PMID- 9199230 TI - An investigation into the prevalence of thyroid disease on Kwajalein Atoll, Marshall Islands. AB - The prevalence of thyroid nodules and thyroid cancer was studied in the indigenous population residing on Ebeye Island, Kwajalein Atoll, in the Republic of the Marshall Islands. This island, centrally located in the nation, is home to about 25% of the nation's population, many who have migrated there from other atolls. The objective of the study was to obtain thyroid disease rate statistics on as much of the population as possible that was alive during the years of nuclear testing and to test the hypothesis that described a linearly decreasing prevalence of palpable nodules with increasing distance from the Bikini test site. 1,322 Marshallese born before 1965 were given a thyroid examination using neck palpation, fine needle aspiration biopsy, and high resolution ultrasound imaging. Approximately 40% of the total population living on this island who are at risk from exposure to radioactive fallout during the years 1946-1958 were screened. Of that group, 815 were alive at the time of the BRAVO test on 1 March 1954. Two hundred sixty-six people with thyroid nodules were found (32.6%): 132 were palpable nodules (16.2%), and 134 were nodules that could be diagnosed with ultrasound only (15.7%). Prevalence of palpable nodules was particularly high in men and women older than 60 y, in men who were 6 to 15 y of age at the time of the BRAVO test, and in women 1 to 10 y of age at the time of the BRAVO test. In 22 people, the clinical diagnosis was most likely cancer though histopathological evidence was only available from 11 operated cases. Of the 11 operated cases, 10 were cancer. Cancer prevalence was particularly high in those women born between 1944 and 1953 (7/220 = 3.2%), i.e., who were children during the early years of nuclear testing. The Ebeye data showed a marginally significant correlation between palpable nodule prevalence among women and distance to Bikini (r = -0.44, p = 0.06). This report summarizes the clinical findings of the thyroid examinations, the age distributions for nodular disease and cancer, and examines the relationship between prevalence of nodules and present day levels of 137Cs in the environment of each atoll. PMID- 9199231 TI - University of Washington's radioecological studies in the Marshall Islands, 1946 1977. AB - Since 1946, personnel from the School of Fisheries, University of Washington (Applied Fisheries Laboratory, 1943-1958; Laboratory of Radiation Biology, 1958 1967; and Laboratory of Radiation Ecology, since 1967), have studied the effects of nuclear detonations and the ensuing radioactivity on the marine and terrestrial environments throughout the Central Pacific. A collection of reports and publications about these activities plus a collection of several thousand samples from these periods are kept at the School of Fisheries. General findings from the surveys show that (1) fission products were prevalent in organisms of the terrestrial environment whereas activation products were prevalent in marine organisms; (2) the best biological indicators of fallout radionuclides by environments were (a) terrestrial-coconuts, land crabs; (b) reef-algae, invertebrates; and (c) marine-plankton, fish. Studies of plutonium and americium in Bikini Atoll showed that during 1971-1977 the highest concentrations of 241Am, 2.85 Bq g(-1) (77 pCi g(-1)) and 239,240Pu, 4.44 Bq g(-1) (120 pCi g(-1)), in surface sediments were found in the northwest part of the lagoon. The concentrations in the bomb craters were substantially lower than these values. Concentrations of soluble and particulate plutonium and americium in surface and deep water samples showed distributions similar to the sediment samples. That is, the highest concentration of these radionuclides in the water column were at locations with highest sediment concentration. Continuous circulation of water in the lagoon and exchange of water with open ocean resulted in removal of 111 G Bq y(-1) (3 Ci y(-1)) 241Am and 222 G Bq y(-1) (6 Ci y(-1)) 239,240Pu into the North Equatorial Current. A summary of the surveys, findings, and the historical role of the Laboratory in radioecological studies of the Marshall Islands are presented. PMID- 9199232 TI - The ecosystem study on Rongelap Atoll. AB - During the 1950's and 1960's, the Laboratory of Radiation Biology at the University of Washington carried out an intensive study of this Atoll, which was contaminated with radioactive fallout from the "Bravo shot" in 1954. This study involved many aspects of the environment and the plant and animal life: soils, land plants, marine life, birds, geology and hydrology, and human diets as well. In much of the research, the fortuitiously present radioactive isotopes, especially 137Cs and 90Sr, were tracers. Although the term "ecosystem study" was not in vogue at that time, it is clear that this was an early use of the ecosystem approach. Soil types and their development, the distribution of mineral elements in plants and soils, including predominant radionuclides, distribution and growth of native terrestrial plants in relation to topography and salinity, some aspects of the human diets, micronutrient nutrition of the coconut palm, island and islet development and stability, were given attention in the studies. Some of the findings in the various areas of study will be presented and discussed. PMID- 9199233 TI - Assessment of a radioactive waste disposal site at Enewetak Atoll. AB - The 43 nuclear tests conducted at Enewetak Atoll by the United States between 1948 and 1958 produced close-in fallout that contaminated the islands and lagoon of the atoll with radioactive fission and activation products, and unfissioned nuclear fuel. In 1972, the U.S. government announced that it would conduct a cleanup and restoration operation to return the atoll to the Enewetak people. The radiological cleanup began in 1977 and lasted to 1980 and focused on reducing the concentration of the transuranium elements (238,239,240Pu and 241Am = TRU) in soils on some of the islands that might eventually be used for residence or for subsistence agricultural. The cleanup plan called for relocating soil and some other contaminated debris to Runit Island on the eastern perimeter of the Atoll. Some of the contaminated soil was mixed with cement and the mixture placed below the water level in the Cactus Crater that was formed by a nuclear explosion in 1958. The remainder of the contaminated material was mixed with concrete and placed above ground over the crater in the shape of a dome. A concrete cap was constructed over the dome of soil. Concern has been expressed by the people of Enewetak and by others over the possible aquatic impacts from the radionuclides entombed in the crater. A National Academy of Sciences committee examined the dome and concluded that the containment structure and its contents present no credible health hazard to the people of Enewetak, either now or in the future. The committee suggested that "at least part of the radioactivity contained in the structure is available for transport to the groundwater and subsequently to the lagoon and it is important to determine whether this pathway may be a significant one." Therefore, a surveillance program was started in 1980, in conjunction with other research efforts, to study the radionuclides in samples of fish, groundwater, and lagoon seawater. Our data and conclusions support the findings suggested by the National Academy committee over a decade ago in that any assumption of rapid remobilization of all or any of the dome's transuranics or other radionuclides is an extreme one. Any fear that this structure contains amounts of activity whose release would cause damage to the environment that will result in greater effect on human health is unfounded. PMID- 9199234 TI - Resuspension studies in the Marshall Islands. AB - The contribution of inhalation exposure to the total dose for residents of the Marshall Islands was monitored at occasions of opportunity on several islands in the Bikini and Enewetak Atolls. To determine the long-term potential for inhalation exposure, and to understand the mechanisms of redistribution and personal exposure, additional investigations were undertaken on Bikini Island under modified and controlled conditions. Experiments were conducted to provide key parameters for the assessment of inhalation exposure from plutonium contaminated dust aerosols: characterization of the contribution of plutonium in soil-borne aerosols as compared to sea spray and organic aerosols, determination of plutonium resuspension rates as measured by the meteorological flux-gradient method during extreme conditions of a bare-soil vs. a stabilized surface, determination of the approximate individual exposures to resuspended plutonium by traffic, and studies of exposures to individuals in different occupational environments simulated by personal air sampling of workers assigned to a variety of tasks. Enhancement factors (defined as ratios of the plutonium-activity of suspended aerosols relative to the plutonium-activity of the soil) were determined to be less than 1 (typically 0.4 to 0.7) in the undisturbed, vegetated areas, but greater than 1 (as high as 3) for the case studies of disturbed bare soil, roadside travel, and for occupational duties in fields and in and around houses. PMID- 9199235 TI - A compilation of nuclear weapons test detonation data for U.S. Pacific ocean tests. AB - Prior to December 1993, the explosive yields of 44 of 66 nuclear tests conducted by the United States in the Marshall Islands were still classified. Following a request from the Government of the Republic of the Marshall Islands to the U.S. Department of Energy to release this information, the Secretary of Energy declassified and released to the public the explosive yields of the Pacific nuclear tests. This paper presents a synopsis of information on nuclear test detonations in the Marshall Islands and other locations in the mid-Pacific including dates, explosive yields, locations, weapon placement, and summary statistics. PMID- 9199236 TI - Some reflections on the role of the Scientific Advisory Panel to the Marshall Islands nationwide radiological study. AB - As a consequence of the U.S. Atomic Weapons Testing Program in the Trust Territory of the Pacific, now the Republic of the Marshall Islands, numerous scientists have advised the Marshallese on matters of radiation and radioactive contamination. Some of the previous advice has appeared to vary or conflict resulting in consequent uncertainty for the people. In a new initiative in 1989, the RMI Government engaged a five member multi-disciplinary Scientific Advisory Panel to oversee the assessment of, and to advise on, the radiological status of the entire nation. The formation of the Panel was accompanied by the establishment of a Resident Scientist position, and ultimately a small scientific team and laboratory on Majuro. The nationwide radiological study was conducted using ground survey methods over the period 1990-1994. Tasks undertaken by the Panel included formulating reasonable objectives for the study and attempting to establish effective communication and understanding of issues with political leaders and RMI Government agencies and people, as well as advising on and monitoring the scientific integrity of the study itself. The attempt was also made to initiate investigations to address matters of concern that emerged. The problem was faced of providing not only technical guidance on radioactivity and radiation measurements, but also explaining the significance of measured values and concepts, such as risk and probability of health effects to a diverse but nontechnical audience, generally across cultural and language barriers. The experience of the Panel in providing advice and guidance to the Republic of the Marshall Islands, while unique in many ways, parallels the difficulties experienced elsewhere in communicating information about risks from radiation exposure. PMID- 9199237 TI - Beyond linearity. PMID- 9199238 TI - Plutonium distribution in the environs of Rocky Flats. PMID- 9199239 TI - Accounting for random error in radon exposure assessment. PMID- 9199240 TI - Localization and intron usage analysis of the human CPT1B gene for muscle type carnitine palmitoyltransferase I. AB - We isolated and sequenced cDNA and genomic DNA fragments of the human CPT1B gene, encoding muscle type camitine palmitoyltransferase I. A recombinant P1 phage containing CPT1B was mapped to chromosome 22qter by fluorescent in situ hybridization. This finding supports the concept that 'liver type' and 'muscle type' isoforms of CPT I are encoded by different loci at separate chromosomal positions. Analysis of CPT1B cDNA sequences revealed the presence of an untranslated 5' exon and differential processing of introns 13 and 19. The alternative splicing of intron 13 causes an in-frame deletion leading to a 10 amino acid residues smaller protein. Using different splice acceptor sites, intron 19 is spliced in the majority of cases, but 4 out of 14 sequenced CPT1B 3' cDNA clones contain part of intron 19 in stead of exon 20. We found that differential polyadenylation is the mechanism behind the existence of these alternative 3' CPT1B mRNA forms. PMID- 9199241 TI - A new functional isoform of the human CRF2 receptor for corticotropin-releasing factor. AB - We have identified the human counterpart of the corticotropin-releasing factor receptor subtype 2beta. Its functional response to human urocortin was demonstrated after stable expression in HEK-293 cells. The receptor was also shown to bind sauvagine, corticotropin-releasing factor and urocortin. In contrast to rodents, the human CRF(2beta) receptor is only weakly expressed in heart and skeletal tissues, where the CRF(2alpha) isoform is predominant. Moreover, we have identified additional mRNAs of the CRF(2beta) type which are probably a consequence of aberrant splicing events. PMID- 9199242 TI - Cloning and expression analysis of mouse Cclp1, a new gene encoding a coiled-coil like protein. AB - Here we describe the nucleotide sequence and expression pattern of a novel gene termed Coiled-coil-like protein 1 (Cclp1). A 2646bp open reading frame encodes a 882 amino acid protein with a predicted coiled-coil domain at the amino terminus. Cclp1 is expressed in a variety of adult tissues and during different stages of embryogenesis. The broad expression pattern suggests a general cellular function of CCLP1. PMID- 9199243 TI - Structural analysis of a bovine arginine tRNA(CCG) gene. AB - A bovine genomic clone containing a 17.4-kb DNA fragment was isolated and found to contain a solitary arginine tRNA gene with an anticodon of CCG that has a 100% identity to its cognate tRNA. This arginine tRNA gene, symbolized as TRR4, has a characteristic internal split promoter and a typical termination site for RNA polymerase III. The tRNA gene was transcribed in vitro by RNA polymerase III using a HeLa cell-free extract to yield a mature-sized tRNA product. The gene was mapped to bovine chromosome 19 using a panel of bovine-rodent somatic cell hybrid DNAs. PMID- 9199244 TI - Isolation of a novel cDNA whose corresponding mRNA is accumulated in growth arrested confluent but not in growing sub-confluent rat 3Y1 cells. AB - Four cDNA fragments, whose corresponding mRNAs were accumulated in growth arrested confluent but not in growing sub-confluent rat 3Y1 cells, were isolated by the mRNA differential display method. Sequencing of the four cDNA fragments indicated that one of them was highly homologous to 3' untranslated DNA region of mouse growth-arrest specific1 cDNA, while the other three (named confluent 3Y1 cell-associated No. 1 (cca1), cca2 and cca3) were novel. Of the three novel cDNAs, we presented some characteristics of cca2 cDNA: the cDNA consisted of 1795 nucleotides with a deduced open reading frame (ORF) encoding a protein of 338 amino acids, which showed a 35% identity with rat type IV 3beta-hydroxysteroid dehydrogenase. The CCA2 protein, with a molecular weight of 38 kDa, was accumulated in 3Y1 cells under growth-arrest conditions. PMID- 9199245 TI - Molecular cloning of cDNA encoding the c-kit receptor of Shiba goats and a novel alanine insertion specific to goats and sheep in the kinase insert region. AB - The complete open reading frame (ORF) of the c-kit cDNA was cloned from a cerebellar cDNA library of the Shiba goat (Capra hircus var Shiba) with the dominant black-eyed white phenotype. The analysis of the deduced amino acid sequence revealed the presence of a single amino acid insertion (alanine) in the kinase insert (KI) region. While the newly found alanine insertion is not correlated with the coat color phenotype of goats, it appears to be characteristic of the c-kit genes in goats and sheep. Although the biological significance of the insert remains to be investigated, its phylogenetically limited distribution will provide us with a useful and interesting tool to analyze the problems of evolution of sheep and goats in bovidae. PMID- 9199246 TI - The sequence of a cDNA encoding functional murine C1-inhibitor protein. AB - The murine C1-inhibitor protein is 482 amino acids long. It consists of an N terminal domain of 118 amino acids rich in proline and threonine and a serpin domain. The N-terminal domain has 39% identity with the corresponding regions of human and bovine C1 inhibitor. PMID- 9199247 TI - Identification and analysis of the rpoS-dependent promoter of katE, encoding catalase HPII in Escherichia coli. AB - The rpoS gene of Escherichia coli encodes an alternative sigma factor of RNA polymerase sigma38 (or sigma(s)) that is required for transcription of katE encoding catalase HPII. The transcription start site of the single katE transcript identified by ribonuclease protection has been determined by primer extension analysis to be either 53 or 54 bp (depending on the strain used) upstream of the open reading frame. A series of promoter fragments were constructed and fused to lacZ to confirm the start site location. A - 10 sequence similar to that found in other sigma70- and sigma38-dependent E. coli promoters was identified 8 or 7 bp upstream of the start site but a sigma70-dependent -35 sequence was not evident. PMID- 9199248 TI - Sequencing of the human vascular endothelial growth factor (VEGF) 3' untranslated region (UTR): conservation of five hypoxia-inducible RNA-protein binding sites. AB - Vascular endothelial growth factor (VEGF) is a potent angiogenic factor whose mRNA expression is induced by hypoxia. This induction is due in large part to an increase in the stability of its mRNA. The RNA sequences and cognate proteins responsible for this increased stability with hypoxia are not well understood. In order to identify regions of functional importance in the 3'UTR of VEGF mRNA, we have sequenced the human VEGF 3'UTR and compared it to the rat sequence. Overall sequence homology was 82% with complete conservation of all four potential polyadenylation signals and both nonameric instability elements. Five hypoxia inducible RNA protein-binding (HI-RPB) sites were identified by RNA electromobility shift assay (EMSA) in the human and rat genes. EMSA and competition studies suggest that these sites bind a similar or related protein complex. On average, the five sites were 95% conserved at the nucleotide level between the rat and corresponding human sequence. This conservation taken together with several previously described, independent correlations between the presence of these RNA-protein complexes and an increase in VEGF mRNA stability suggest an important functional role for these sites in mediating hypoxia inducible VEGF mRNA stability. PMID- 9199249 TI - Nuclear genes for cytochrome c oxidase. PMID- 9199250 TI - Green fluorescent protein as a reporter of gene expression in transgenic mice. AB - We used the green fluorescent protein (GFP) from the jellyfish Aequorea victoria as a reporter of gene expression in transgenic mice. The GFP coding sequence was placed under the control of the human hemopexin and the mouse beta1 integrin promoter that were previously studied in transgenic mice using the lacZ reporter gene. We showed that GFP has a higher degree of sensitivity compared to the lacZ reporter gene allowing to identify cells with low and otherwise undetectable beta galactosidase activity. Thus we showed the potentiality of GFP in replacing lacZ as a reporter gene to investigate promoter mapping and gene regulation in transgenic mice. PMID- 9199251 TI - A novel type of dimerization motif, related to leucine zippers, is present in plant homeodomain proteins. AB - Sunflower HAHR1 is a homeodomain protein presumably involved in some aspects of root development. In the present work, we have studied the oligomerization properties of HAHR1. A protein containing the entire homeodomain plus adjacent C terminal sequences (amino acids 86-325) behaves as a dimer in gel filtration experiments. When a fragment C-terminal to the homeodomain (amino acids 151-263) is fused to the N-terminal domain of the lambda phage repressor, it is able to confer binding efficiency to this domain, as judged by protection from lambda superinfection and repression of beta-galactosidase expression under the control of the P(R) promoter. A smaller fragment (amino acids 151-184) confers only conditional repression. GSH transferase fusion proteins containing the entire homeodomain of HAHR1 plus the above-mentioned adjacent sequences bind with similar efficiency a mixture of oligonucleotides selected from a random population. The smaller protein, however, loses its binding capacity when separated from the GSH transferase moiety. Retention of a labelled HAHR1 protein synthesized in vitro by GSH transferase fusions containing different protein fragments adjacent to the homeodomain and bound to GSH agarose suggests that a portion from amino acids 151-263 is required for efficient interaction. The results obtained indicate that HAHR1 interacts with DNA as a dimer and that its dimerization domain is located immediately C-terminal to the homeodomain. We define two regions, the first of which confers non-efficient dimerization; this region would be stabilized by the presence of the second one through putative mutual interactions. A similar motif is present in other related plant homeodomain proteins. PMID- 9199252 TI - Identification of processes that influence negative supercoiling in the human c myc gene. AB - DNA elements with sequences suitable for Z-DNA formation are found frequently at various positions in chromatin. Z-DNA formation in these sequences depends largely on the level of local negative supercoiling. We can use binding of a Z DNA specific antibody at low concentrations in metabolically active permeabilized nuclei to detect naturally occurring Z-DNA formation. Previously we identified three sequence elements in the human c-myc gene that adopt the Z-DNA conformation in the transcribed gene. The three elements are found far upstream (Z1), close to the main transcription start site (Z2) and in the first intron (Z3). Here we measure the persistence of Z-DNA at these three sites under the influence of various metabolic inhibitors. This provides some insight into the varying levels of negative supercoiling. alpha-Amanitin, an inhibitor of transcription, reduced the persistence of Z-DNA in all three elements. Aphidicolin, an inhibitor of replication, increased the persistence of Z-DNA in one element without significantly influencing the other two elements. When camptothecin an inhibitor of topoisomerase I was added in the presence of alpha-amanitin, the persistence of Z-DNA was extended in all three elements. However, in the presence of aphidicolin no effect of camptothecin on Z-DNA formation was observed. PMID- 9199253 TI - Molecular cloning of cDNAs for mouse low-molecular-weight and high-molecular weight prekininogens. AB - We isolated cDNAs encoding low-molecular-weight (L-) and high-molecular-weight (H ) prekininogens from a mouse liver cDNA library using rat T-kininogen cDNA and rat H-kininogen cDNA respectively, as probes. The signal peptide, the heavy chain, and the bradykinin moiety, which are common between the two prekininogens, consist of 20, 359, and 9 amino acids, respectively, while the light chains of the L- and H-prekininogens are composed of 44 and 273 amino acids, respectively. All 19 cysteine residues present in both mouse prekininogens are located at the same positions relative to those of human origin. The light chain of H prekininogen contains a characteristic 15-repeated His-Gly sequence and a conserved sequence for binding prekallikrein or factor XI. Northern blotting or reverse transcription-polymerase chain reaction followed by Southern blotting using mouse L- and H-kininogen cDNAs demonstrated that both L- and H-kininogens are predominantly expressed in the liver and kidney. L-Kininogen mRNA was also expressed in other tissues, such as the adrenal gland, brain, spinal cord, testis, lung, heart, and skin, while levels of H-kininogen mRNA in these tissues were too low to detect, suggesting that L-kininogen is synthesized in various tissues of mouse, while H-kininogen is exclusively synthesized in the liver and kidney. A genomic Southern blot using H-prekininogen cDNA revealed that the L- and H-prekininogen mRNAs in mouse are probably encoded by a single gene, as is the case in both human and bovine. PMID- 9199255 TI - Prediction of severe coronary artery disease by combined rest and exercise radionuclide angiocardiography and tomographic perfusion imaging with technetium 99m-labeled sestamibi: a comparison with clinical and electrocardiographic data. AB - BACKGROUND: The purpose of this study was to compare the incremental value of clinical information, electrocardiographic data, myocardial perfusion imaging, and radionuclide angiography for predicting severe coronary artery disease at a single testing interval. Clinical information, treadmill exercise studies, radionuclide angiography, and myocardial perfusion imaging are important predictors of severe coronary artery disease. However, the relative and absolute diagnostic importance of each of these methods has not been addressed at a single testing interval. METHODS AND RESULTS: A same-day rest/treadmill exercise perfusion and function study was performed in 167 patients within 90 days of coronary angiography. A multivariable regression model was used to assess the independent informational content of these predictors. Clinical and electrocardiographic data were related strongly to the presence of severe coronary artery disease (chi2 = 12.2 and p < 0.001; chi2 = 11.8 and p < 0.001, respectively). Combined perfusion and functional studies contributed 31% of the diagnostic information beyond that provided by clinical and electrocardiographic data alone (p < 0.05). CONCLUSIONS: These data demonstrate that combined studies of myocardial perfusion and left ventricular function are able to improve prediction of the extent of coronary artery disease, even when clinical and electrocardiographic data are also available. PMID- 9199256 TI - Prognostic value of persistent thallium-201 defects that become reversible after reinjection in patients with chronic myocardial infarction. AB - BACKGROUND: The presence of defects at stress-redistribution thallium-201 scintigraphy is related to a higher risk of cardiac events. However, the prognostic value of defects that become reversible after reinjection is not known. In this study we evaluated the prognostic contribution of stress redistribution-reinjection with special regard to 3-hour fixed defects that become reversible after reinjection. METHODS AND RESULTS: We studied 122 patients with chronic myocardial infarction (>2 months) and suspected or known residual ischemia, with stress-redistribution-reinjection planar scintigraphy. Thallium scans were analyzed by three observers (three segments per view, 5-point score) and classified as normal, fixed, and reversible. The lung/heart ratio was also calculated. At a median follow-up of 47 months, 10 patients had hard events (four deaths and six myocardial infarctions) (group I), 12 patients had unstable angina (group II), 12 patients underwent planned coronary artery bypass grafting or percutaneous transluminal coronary angioplasty (group III), and 86 patients had no events (group IV). The presence of fixed defects that became reversible after reinjection did not identify patients at higher risk. The number of reversible defects at 3 hours was significantly higher only in patients who underwent revascularization. Unstable angina was not predicted by any scintigraphic pattern. The variables that were statistically related to hard events by univariate analysis were increased lung uptake, reversible cavity dilation, and the number of fixed defects that remained fixed after reinjection. By Cox multivariate analysis, the strongest predictor of hard events was the presence of more than three fixed defects that remained fixed after reinjection as a marker of irreversible myocardial damage. CONCLUSIONS: (201)Tl reinjection is a useful approach for not only detecting viable myocardium but also risk stratification in patients with chronic myocardial infarction. PMID- 9199257 TI - Enhanced accuracy of defect detection by myocardial single-photon emission computed tomography with attenuation correction with gadolinium 153 line sources: evaluation with a cardiac phantom. AB - BACKGROUND: Photon attenuation is a major cause of artifacts on single-photon emission tomographic imaging. METHODS AND RESULTS: To study a new method to perform photon attenuation correction (AC), we used a cardiac phantom filled with (99m)Tc and imaged it (1) without extrinsic attenuation or defects, (2) with extrinsic attenuation but without defects, (3) without extrinsic attenuation but with defects involving 10% of the myocardial volume and activity ranging from 0% to 75% of maximum, and (4) with both extrinsic attenuation and defects. Transmission and emission images acquired with a dual-head single-photon emission computed tomographic system with 153Gd line sources were processed by iterative maximum-likelihood-expectation-maximization and were evaluated both qualitatively and quantitatively. The small defects were readily identified both before and after AC. Mean count activity (percent of maximal activity) of the segments with overlying extrinsic attenuation but without defects was only 56% +/- 4% without AC but increased to 86% +/- 4% with AC (p < 0.0001). Without AC, the count activities in the defects with overlying extrinsic attenuation were lower than the actual defect activities, but AC resulted in better approximation of actual defect activities in all but the most severe (0% tracer activity) defects. CONCLUSION: This new AC method provided an improved estimation of actual myocardial count activity. Even small defects with mild reduction in tracer activity were still identifiable after AC. PMID- 9199258 TI - Comparison of arbutamine stress 99mTc-labeled sestamibi single-photon emission computed tomographic imaging and echocardiography for detection of the extent and severity of coronary artery disease and inducible ischemia. AB - BACKGROUND: Arbutamine is a new synthetic catecholamine developed specifically for pharmacologic stress testing. METHODS AND RESULTS: We investigated 39 patients undergoing coronary arteriography to compare arbutamine stress (99m)Tc labeled sestamibi single-photon emission computed tomographic imaging and echocardiography for detection of the extent and severity of coronary artery disease and inducible ischemia. Rest and stress studies were analyzed blindly according to a 12-segment left ventricular model for both techniques. Each segment was graded according to severity of wall thickening abnormality and perfusion defect (1 = normal to 4 = severe). Total perfusion defect and wall thickening scores were calculated at peak stress and the difference in scores between stress and rest (delta perfusion defect; delta wall thickening) were used as indexes of inducible ischemia. Twenty-one patients had multivessel disease, nine had single-vessel disease, and nine had normal coronary arteries. Diagnostic accuracies for the detection of coronary artery disease for single-photon emission computed tomographic imaging and echocardiography were 95% and 92%, respectively. Extent and severity of coronary artery disease indicated by a peak stress perfusion defect score of 26 +/- 6.4 and wall thickening score of 25.1 +/- 8.4 were similar, and there was no significant difference in the delta perfusion defect and delta wall thickening scores of 8.7 +/- 5.5 and 10.4 +/- 7.1, respectively. Segmental concordance rates for the detection of coronary artery disease and inducible ischemia were 74% (K = 0.47; confidence interval 0.39 to 0.55) and 74% (kappa = 0.42; confidence interval 0.34 to 0.51), respectively. Regional concordance for coronary artery disease was 84% (kappa = 0.68; confidence interval 0.51 to 0.84). Where discordance was present, there was a greater prevalence of perfusion abnormality compared with wall thickening abnormality. CONCLUSION: Arbutamine stress single-photon emission computed tomographic imaging and echocardiography provide largely equivalent and accurate pathophysiologic information for the evaluation of coronary artery disease and inducible ischemia. PMID- 9199259 TI - Planar imaging of 99mTc-labeled (bis(N-ethoxy, N-ethyl dithiocarbamato) nitrido technetium[V]) can detect resting ischemia. AB - BACKGROUND: (99m)Tc-labeled (bis(N-ethoxy, N-ethyl dithiocarbamato) nitrido technetium(V)) ([99m]TcN-NOET) is a new lipophilic, neutral-charge cardiac perfusion imaging agent that demonstrates apparent redistribution in animal models and humans. The purpose of this study was to determine whether the kinetics of (99m)TcN-NOET are suitable for the detection of resting ischemia. METHODS AND RESULTS: Microspheres were injected at baseline and simultaneously with (99m)TcN-NOET after a 90% reduction in resting flow in the left circumflex coronary artery in six open-chest canine experiments. The relationship of flow and activity early after injection was determined in one experiment by termination at 10 minutes. The flow ratio (left circumflex/left anterior descending coronary artery) after stenosis fell significantly (0.87 +/- 0.04 vs 0.46 +/- 0.04; p < 0.05). The end-tissue (99m)Tc ratio (0.78 +/- 0.05) was significantly higher than the flow ratio at injection (0.46 +/- 0.04; p < 0.05), indicating substantial redistribution. In vivo imaging was conducted during 2 hours in five experiments, followed by ex vivo imaging. Myocardial clearance from 10 minutes onward was biphasic in left anterior descending and monophasic in left circumflex coronary arteries. Myocardial clearance from 10 to 60 minutes was delayed in left circumflex (35.5% +/- 8.1%) versus left anterior descending coronary arteries (49.2% +/- 8.6%; p < 0.05). No significant difference was observed from 60- to 120-minute clearance. Five of five experiments demonstrated initial defects and complete fill-in at 90 to 120 minutes by qualitative assessment. Quantitation of ex vivo images confirmed significant redistribution. CONCLUSIONS: Resting ischemia caused by moderate to severe stenosis can be detected on scans with (99m)TcN-NOET. Redistribution was near complete in this model by 90 to 120 minutes. (99m)TcN-NOET is a promising new agent for the detection of coronary artery disease in viable myocardium and warrants further investigation. PMID- 9199260 TI - Maximizing radiotracer delivery to experimental atherosclerotic lesions with high dose, negative charge-modified Z2D3 antibody for immunoscintigraphic targeting. AB - BACKGROUND: Two factors that directly affect target/background ratio in immunoscintigraphy are the concentration of the antibody bound to the target and the concentration of the antibody in the circulation. High dosages of monoclonal antibody have been reported to be more efficacious in visualization of tumors. Although administration of a higher dosage of antibody increases the absolute target accumulation of the radiotracer, it also increases the background activity, which may offset this advantage. Negative charge-modified antibodies carry high specific radioactivity to the target sites without significantly increasing the background activity. Therefore we investigated whether higher dosages of negative charge-modified antibody can be used to improve imaging of experimental atherosclerotic lesions. METHODS AND RESULTS: Experimental atherosclerotic lesions were produced in 16 New Zealand White rabbits by balloon deendothelialization of the infradiaphragmatic aorta and hyperlipidemic diet for 12 weeks. Negative charge-modified Z2D3 antibody F(ab')2 specific for an antigen on proliferating smooth muscle cells of human atheroma labeled with (111)In was used for imaging experimental atherosclerotic lesions either at high (100 to 125 microg) or low (25 to 50 microg) dosages. A lower dosage of Z2D3 was labeled with 507 +/- 29.5 microCi (25 to 50 microg) (111)In label, compared with 2.9 +/- 0.24 mCi (100 to 125 microg) for the higher dosage. Although noninvasive visualization of atherosclerotic lesions was possible in all animals at 24 hours, high antibody dose allowed unequivocal visualization of the lesion as early as 3 hours after intravenous administration of the antibody. Eight animals were killed at 24 hours and the remaining eight animals at 48 hours. Mean radioactivity dose delivered per gram of lesion with the low-dose protocol at 24 hours was 0.46 +/- 0.09 microCi, which remained essentially unchanged at 48 hours (0.37 +/- 0.09 microCi; p = 0.51). With the high-dosage protocol, the total radioactivity (dose) per gram uptake in the lesion increased by about eightfold (3.49 +/- 0.58 microCi; p = 0.002) at 24 hours and was sixfold higher at 48 hours (2.21 +/- 0.45 microCi; p < 0.02). CONCLUSIONS: The study demonstrated that the increase in the dosage of negatively charge-modified antibody allows a very high delivery of specific radioactivity to the target, which in turn enables early visualization of experimental atherosclerotic lesions. PMID- 9199261 TI - Understanding Fourier space and filter selection. PMID- 9199262 TI - "We now join the freeway (information superhighway), which is already in progress". PMID- 9199263 TI - Improving our image. PMID- 9199264 TI - Noninvasive detection of acute heart rejection: the quest for the perfect test. PMID- 9199265 TI - Scintigraphic demonstration of myocardial sarcoidosis: the added value of single photon emission computed tomography. PMID- 9199266 TI - Expression of syndecan-1 in rabbit neointima following de-endothelialization by a balloon catheter. AB - Enrichment of proteoglycans is prominent in early atherogenesis, contributing not only to SMC migration and proliferation, but also to low density lipoprotein retention. A family of integral cell membrane proteoglycans termed syndecans has recently been recognized. Among syndecans, syndecan-1, the first isolated member, has received most research attention. In this study, we examined the expression of syndecan-1 in rabbit aorta and aortic neointima, developed in response to a balloon catheter-induced de-endothelialization. The tissues were processed for Northern blot analysis, in situ hybridization, immunohistochemical staining and immunoblotting. Our results indicate that in normal aorta, the signal for syndecan-1 is weak. However, arterial injury induces syndecan-1 expression at both mRNA and protein levels. The presence of syndecan-1 in the neointimal tissue is persistent, prominent even at the 12th week after injury. Syndecan positive cells are distributed in the whole layer of the neointima, but are not visible in the underlying media. The presence of syndecan-1 in arterial neointima suggests a novel means of mediating interactions between neointimal cells and various agents, including extracellular matrix components, growth factors and lipoproteins. PMID- 9199267 TI - Surface expression of low density lipoprotein receptor in EBV-transformed lymphocytes: characterization and use for studying familial hypercholesterolemia. AB - The objectives of the present study were to characterize the surface expression of low density lipoprotein receptor (LDL-R) in Epstein-Barr virus transformed lymphocytes (EBV-L) and to determine the applicability of the cellular system for the study of familial hypercholesterolemia. The EBV-L subsets and LDL-R expression were determined by immuno-cytofluorimetry. The LDL-R expression in EBV L which consisted of mostly B cells was no different among antigenic subsets. EBV L cultured in lipoprotein deficient serum demonstrated a 9.3-fold higher LDL-R expression than primary lymphocytes. Lovastatin caused an additional 1.9-and 1.4 fold increase in EBV-L and primary lymphocytes respectively. This difference in lovastatin response is statistically significant (paired t-test, P < 0.001). 54% of the high LDL-R expression in EBV-L was related to the changes in proliferation measured as stimulation index (SI). LDL and lovastatin modulated the LDL-R expression without affecting SI. FH subjects demonstrated 2% (homozygote, n = 1) and 44.6 +/- 12.3% (heterozygotes, n = 35) in LDL-R expression of controls (n = 30). This maintenance of the FH phenotype and the intrinsically high LDL-R expression in EBV-L make the cellular system suitable for the study of FH as well as the regulation of LDL-R. PMID- 9199268 TI - Alterations in plasma lipids, lipoproteins and high density lipoprotein subfractions in peripheral arterial disease. AB - The concentrations of the major lipoprotein classes and of high density lipoprotein (HDL) subfractions in 63 male patients with arteriosclerosis of the lower limbs (claudication) were determined and compared with values from 63 healthy controls. The patients with peripheral arterial disease (PAD) had reduced levels of total HDL-cholesterol and HDL2b of large particle size, increased levels of small HDL3c particles and a high ratio of total plasma-cholesterol to HDL-cholesterol (coronary risk factor). The PAD patients, however, had lower levels of low density lipoprotein (LDL)-cholesterol but higher concentrations of very low density lipoprotein (VLDL)-cholesterol and plasma triglyceride than healthy subjects. This study therefore suggests that in PAD, the protective effect of HDL may be more important than the atherogenic effect of LDL. It further suggests that while HDL-cholesterol HDL2b and the ratio of total plasma cholesterol to HDL-cholesterol may provide valid indices for identifying individuals at risk of PAD, other factors, such as LDL and total cholesterol, may not provide such an appropriate risk indicator. PMID- 9199269 TI - Augmented Ca2+ influx is involved in the mechanism of enhanced proliferation of cultured vascular smooth muscle cells from spontaneously diabetic Goto-Kakizaki rats. AB - To investigate whether augmented calcium influx is involved in the mechanism of the enhanced proliferation of vascular smooth muscle cells (VSMCs) in diabetes, we studied the association between proliferation and cytosolic free calcium concentration ([Ca2+]i) in cultured aortic VSMCs from spontaneously diabetic Goto Kakizaki (GK) and Wistar rats. Serum, angiotensin II and Bay K 8644, a voltage dependent Ca2+ channel (VDC) agonist, stimulated the proliferation of VSMCs; the magnitude was greater in VSMCs from GK than Wistar rats. VDC blockers, verapamil and nicardipine, inhibited Bay K 8644-induced cell proliferation, and the difference in the proliferation of VSMCs between GK and Wistar rats disappeared. Angiotensin II-induced proliferation was only partially inhibited by VDC blockers, and enhanced proliferation of GK-VSMCs was still observed. Bay K 8644 and angiotensin II increased [Ca2+]i, and the increase was augmented in GK-VSMCs. Bay K 8644-induced [Ca2+]i increase was completely inhibited by pretreatment with verapamil or removal of extracellular Ca2+, suggesting that VDC is associated with this increase. Although angiotensin II-induced [Ca2+]i increase was not affected by verapamil, removal of extracellular Ca2+ slightly but significantly attenuated angiotensin II-induced [Ca2+]i increase, suggesting that VDC blocker insensitive receptor-activated Ca2+ influx is involved. These results indicate that augmented Ca2+ influx via VDC and a receptor-activated pathway may be involved in the mechanism of the enhanced proliferation of VSMCs from GK rats. PMID- 9199270 TI - The role of Fas/APO 1 and apoptosis in the development of human atherosclerotic lesions. AB - The possibility that Fas/APO 1 is involved in the apoptosis of advanced human coronary atherosclerosis was examined in the present study. Coronary arteries with atherosclerosis were obtained from human hearts with chronic ischemic heart disease at cardiac transplantation. Normal vessels were used as controls. Fas/APO 1 was detected by immunohistochemistry with a monoclonal antibody. Apoptotic cells were stained in situ by terminal deoxynucleotidyl transferase mediated-dUTP nick end labeling (TUNEL) and DNA fragmentation into oligonucleosomes was checked by gel electrophoresis. Bcl-2, an antiapoptotic oncoprotein, was detected by immunohistochemistry and Western blot. Apoptotic cells were present in the neointima in all stages of atherosclerosis, and in intraplaque small vessels. In initial lesions, only a few cells were undergoing apoptosis. By contrast, in advanced lesions, many cells were found to undergo apoptosis. Apoptosis was further confirmed by genomic DNA analysis using gel electrophoresis. Apoptotic cells were either smooth muscle cells or macrophages, but also endothelial and blood borne cells. Fas/APO 1 was present in foam cells. Most of the Fas/APO 1 positive cells were stained for the macrophage marker CD68 and for alpha-smooth muscle actin in serial sections. Several anti-Fas/APO 1 positive foam cells were revealed to undergo apoptosis by double staining. Bcl-2 was detected in Fas/APO 1 expressing plaques. A number of CD3-positive T-lymphocytes were found around foam cells expressing Fas/APO 1. This data suggests that Fas/APO 1 regulated apoptosis is involved in the development of advanced human atherosclerotic lesions and that it probably determines the amount of tissue mass in the diseased vessels. PMID- 9199271 TI - Increased proliferation of explanted vascular smooth muscle cells: a marker presaging atherogenesis. AB - The JCR:LA-cp homozygous cp/cp corpulent rat is genetically predisposed to develop atherosclerosis evident after 9 and 18 months of age in males and females and to manifest metabolic derangements resembling those seen in type II diabetes in humans (hyperinsulinemia, insulin resistance, hyperglycemia and hypertriglyceridemia). The present study was undertaken to determine whether vascular smooth muscle cells (SMCs) explanted from vessels destined to become atherosclerotic later in life exhibit intrinsic properties ex vivo that presage atherogenesis to provide a means for evaluating promptly intervention designed to modify it. SMCs were cultured from aortic explants of JCR:LA-cp corpulent (cp/cp) and lean control (+/+) rats of 4, 5, 6, and 9 months of age. Compared with SMCs from controls, SMCs from cp/cp rats exhibited increased proliferation, higher saturation density, increased augmentation of proliferation in response to selected mitogens and greater adherence to extracellular matrix proteins. The increased proliferative activity ex vivo anteceded by several months the development of atherosclerotic lesions in vivo. Thus, it is a promising marker in assessments of the efficacy of interventions designed to retard or prevent atherosclerosis. PMID- 9199272 TI - Impairment of endothelium-dependent relaxation of rabbit aortas by cigarette smoke extract--role of free radicals and attenuation by captopril. AB - The aim of this study was to examine the effects of the water soluble component of cigarette smoke extract (CSE) on endothelium-dependent relaxation (EDR) of isolated rabbit aortas. The incubation with CSE was found to inhibit EDR in a dose-dependent manner. Co-incubation of the aortic strips with superoxide dismutase (SOD), N-acetylcysteine, glutathione or dimethyl sulfoxide (DMSO), free radical scavengers, attenuated the CSE-induced inhibition of the arterial relaxation. Co-incubation of the strips with captopril (3 mM), an angiotensin converting enzyme inhibitor, also attenuated CSE-induced impairment of vasorelaxation. In parallel experiments using cultured human endothelial cells, CSE suppressed endothelial release of NOx, stable metabolites of nitric oxide (NO). SOD, DMSO and captopril attenuated the suppression of NO production by CSE in association with reduction of free radicals, superoxide anions and hydroxyl radicals, in CSE solution. Neither lactate dehydrogenase release from the cultured endothelial cells nor cell death estimated by trypan blue exclusion test was found after the incubation of the cultured endothelial cells with CSE. The results indicate that free radicals in CSE induce the impairment of EDR, which may be partly due to suppression of NO production and is not due to non-specific cytotoxicity by CSE. Captopril attenuates CSE-induced endothelial dysfunction partly through scavenging free radicals. PMID- 9199273 TI - Abnormalities of serum apo A1 containing lipoprotein particles in patients with primary biliary cirrhosis. AB - Patients with primary biliary cirrhosis (PBC) do not appear to have an increased risk of cardiovascular disease despite elevations in serum cholesterol. Recent evidence has pointed to LpA1 (an apo A1 containing particle which contains apo A1 but not apo A2) in protecting against atherosclerosis. The aim of this study was to investigate apo Al containing particles in the serum of patients with PBC. Lipids and apolipoproteins were measured in 31 patients with PBC (30 females) and 27 control subjects (26 females). Patients were divided into 3 groups: group 1 with bilirubin < 18 micromol/l (n = 17); group 2 with bilirubin > 18 micromol/l (n = 11); and group 3 with end stage liver disease (ESLD, n = 3). As expected group 1 and 2 patients had higher total cholesterol, HDL cholesterol and phospholipids than control subjects. Apo B and apo A1 concentrations were similar to control subjects. However, LpA1 was greatly increased: 0.96 g/l (0.60-1.50), median (range) in group 1 and 1.09 g/l (0.75-1.33) in group 2 versus 0.62 g/l (0.45-0.93) for controls both P < 0.005 and the percentage of total apo A1 in the LpA1 fraction was increased: 54.8% (37.9-63.4) in group 1 and 55.7% (47.8-73.7) in group 2 versus 36.8% (25.1-49.1) for controls, both P < 0.005. Apo A2 concentration was reduced in group 1 0.38 g/l (0.30-0.51) and group 2 0.31 g/l (0.14-0.58) versus controls 0.43 g/l (0.36-0.57), P < 0.05 and P < 0.005 respectively. Patients with ESLD had reduced HDL cholesterol, apo A1, LpA1 and apo A2 compared to controls. These results suggest that PBC is associated with an altered distribution of apo A1 favouring an increased concentration of the protective LpA-I particles. Increased LpA1 concentration may be one of the factors contributing to the paradoxically low incidence of atherosclerosis in PBC patients. PMID- 9199274 TI - Plasma fibrinogen concentration in a Chinese population. AB - Plasma fibrinogen concentration has been shown to be a predictor of major cardiovascular events. Information on plasma fibrinogen amongst Chinese has been scanty. We examined the relationships between plasma fibrinogen concentration and cardiovascular risk factors in 988 chinese subjects who underwent 75 g oral glucose tolerance test for screening for glucose intolerance. The study involved a selected sample with subjects who had an history of gestational diabetes, delivery of big babies (birth weight > or = 4 kg), equivocal plasma glucose concentrations and subjects who were family members of diabetic patients. This was mainly a non-smoking (96.6%), non-drinking (98%) and non-exercising (99%) population of which 87% (n = 855) were female. Among the 988 subjects (age +/- S.D. 36.8 +/- 10.2, range 16-79 years), plasma fibrinogen concentration ranged from 1.40 to 9.90 g/l with a mean of 3.26 +/- 0.93 g/l. On stratification of the subjects into 4 quartiles based on plasma fibrinogen concentrations, we found that increased plasma fibrinogen was associated with older age, higher body mass index (BMI), systolic and diastolic blood pressure (BP), fasting and 2 h plasma glucose (PG), prevalence of diabetes, glycated haemoglobin (HbA1c) and triglyceride (TG) level. After adjustment for age and sex, increased plasma fibrinogen concentration remained associated with higher BMI, systolic BP, 2 h PG and TG level. On multivariate analysis using age, BMI, BP, TG, HbA1c and PG as independent variables, plasma fibrinogen was independently related to plasma TG concentration and HbA1c. With 1 S.D. change in TG concentration and HbA1c, there were 3.7 and 5.2% changes in plasma fibrinogen concentration respectively. These findings emphasize the close relationships between plasma fibrinogen and cardiovascular risk factors, in particular abnormal lipid and glucose metabolism. PMID- 9199275 TI - Mechanisms of enhanced production of PGI2 in cultured rat vascular smooth muscle cells enriched with eicosapentaenoic acid. AB - The present investigation was performed to clarify the effect of EPA on PGI2 production in vitro using cultured rat vascular smooth muscle cells (VSMC). To simulate in vivo conditions, a triacylglycerol (TG) emulsified form of EPA was used. An increase in EPA content was achieved without alteration of arachidonic acid concentration. These experiments clearly demonstrated that co-incubation of EPA-TG increased PGI2 production by cultured VSMC in a dose dependent fashion. Among polyunsaturated fatty acid TG examined (docosahexaenoic acid, linoleic acid, oleic acid and EPA), only EPA-TG was effective. Cyclooxygenase (COX) was activated, but neither phospholipase A2 nor PGI2 synthase activity was changed. EPA treatment did not alter the amount of COX-1 and COX-2 protein in VSMC. Addition of antioxidants, such as butylated hydroxytoluene or vitamin E, decreased MDA levels in the medium and cells and reversed the enhanced PGI2 production in EPA rich-VSMC. Therefore, the high polyunsaturation of EPA could generate low levels of lipid peroxides and thereby lead to activation of COX and an increased PGI2 production. Although EPA increased PGI2 production, only a negligible amount of PGI3 was produced by rat aortic tissues. Enhanced production of PGI2 might contribute to the anti-atherogenic effect of EPA. PMID- 9199276 TI - Atherogenic lipoprotein changes in the absence of hyperlipidemia in patients with chronic renal failure treated by hemodialysis. AB - We compared plasma lipid and lipoprotein parameters between 210 chronic renal failure patients treated by hemodialysis and 223 age- and sex-matched healthy control subjects to examine whether atherogenic lipoprotein changes were present in hemodialysis patients in the absence of hyperlipidemia. The hemodialysis group showed higher levels of plasma triglycerides, very low density lipoprotein (VLDL) cholesterol, and intermediate density lipoprotein (IDL) cholesterol and a lower level of high density lipoprotein (HDL) cholesterol. Low density lipoprotein (LDL) cholesterol of the hemodialysis group was not elevated but their LDL was significantly more triglyceride-enriched than that of controls. Subjects were then divided into five categories according to their plasma triglyceride levels at an interval of 50 mg/dl, and comparison was made between the two groups in the same range of plasma triglycerides. Hemodialysis patients again showed higher levels of VLDL- and IDL-cholesterol, and lower levels of HDL-cholesterol than the control group even in the plasma triglycerides-matched comparisons. Similarly, higher VLDL- and IDL-cholesterol levels in hemodialysis patients were significant in plasma total cholesterol-matched subgroup comparisons. Multiple regression analysis indicated that the relationship between plasma lipid concentrations and individual lipoprotein levels were substantially altered in uremic state. The 95th percentile level of IDL-cholesterol in the nonuremic controls was 15 mg/dl, and 45% of hemodialysis patients exceeded this level. Decreased HDL-cholesterol levels < or = 35 mg/dl were seen in 6% of the control and 38% of the hemodialysis group. Elevated IDL-cholesterol and decreased HDL-cholesterol were persistently found in hemodialysis patients with normal lipid levels. It is concluded that hemodialysis patients exhibited more atherogenic lipoprotein profile than nonuremic subjects with comparable levels of plasma triglycerides and total cholesterol. Especially, increased IDL- and decreased HDL-cholesterol levels in hemodialysis patients persisted even at very low levels of plasma lipids. Since elevated IDL and decreased HDL-cholesterol are implicated in the progression of atherosclerosis, these findings are of clinical importance in the diagnosis of lipoprotein disorder in chronic renal failure. PMID- 9199277 TI - Reduction of oxysterol levels up-regulates HMG-CoA reductase activity in rat liver. AB - Cholesterol regulates hepatic 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase activity by feedback inhibition. It has been suggested that oxidized derivatives of cholesterol (oxysterols) play an important role, as an intracellular mediator, in the feedback inhibition of cholesterol biosynthesis. We, therefore, investigated the role of intracellular oxysterols in the regulation of HMG-CoA reductase activity. Rats were fed with food (control), cholesterol, clofibrate as a potentiator of the microsomal monooxygenase cytochrome P-450 enzyme system, ketoconazole as a strong inhibitor of the system, or butylated hydroxytoluene (BHT) as an antioxidant. We analyzed and compared hepatic microsomal oxysterol levels among the groups. The results of this study indicated that the oxysterol level, especially 7beta-hydroxycholesterol and 7 ketocholestrol, in the liver was lowered by the administration of ketoconazole and BHT, and HMG-CoA reductase activity was increased in response to these agents. However, there was no change in the HMG-CoA reductase activity, after the administration of clofibrate. We conclude that reduced levels of oxysterol may release the inhibitory effect on the HMG-CoA reductase enzyme and lead to up regulation of the enzyme. PMID- 9199278 TI - Apolipoprotein E polymorphism has no independent effect on plasma levels of lipoprotein(a). AB - Previous studies show conflicting results concerning an influence of apolipoprotein E (apo E) phenotype on lipoprotein(a) (Lp(a)) plasma levels. We speculated that it is not the apo E phenotype itself but rather its effect on plasma lipid concentrations that might influence Lp(a) levels. In 1562 subjects concentrations of triglycerides, LDL-cholesterol and Lp(a) were measured by standard laboratory methods. Apo(a) and apo E isoforms were determined by sodium dodecyl sulfate gel electrophoresis and isoelectric focusing, respectively, followed by immunoblotting. An univariate analysis revealed a significant influence of apo(a) isoforms, apo E phenotype, triglycerides and LDL-cholesterol on Lp(a) plasma levels (ANOVA: P < 0.001, P < 0.02, P < 0.001 and P < 0.001, respectively). In a multivariate analysis, however, the influence of the apo E phenotype was no longer significant (P>0.10), whereas apo(a) isoforms, LDL cholesterol quintiles and triglyceride quintiles explained 29.2, 2.8 and 1.0% of the variation of the Lp(a) levels (for all three variables: P < 0.001). We conclude that apo E polymorphism does not exert an independent effect on Lp(a) concentrations. Any influence is mediated through the effect of apo E polymorphism on plasma lipids. PMID- 9199279 TI - Influence of two apo A4 polymorphisms at codons 347 and 360 on non-fasting plasma lipoprotein-lipids and apolipoproteins in Asian Indians. AB - Apolipoprotein A-IV (apo A-IV, protein; apo A4, gene) is a major constituent of triglyceride-rich and high-density lipoprotein particles and may, therefore, play an important role in lipid metabolism. We studied the distribution of two apo A4 polymorphisms at codons 347 (alleles A and T) and 360 (alleles 1 and 2) in relation to plasma lipoprotein-lipid and apolipoprotein levels in 176 non-fasting male blood donors from New Delhi, Northern India. The frequencies of the T allele at codon 347 and the 2 allele at codon 360 were 0.12 and 0.03 respectively. Carriers of the T allele (AT and TT genotypes) had significantly lower plasma total cholesterol (P = 0.04) and low density lipoprotein (LDL)-cholesterol (P = 0.02) levels than individuals homozygous for the A allele (AA genotype). The codon 347 polymorphism explained 2.2 and 2.6% of the phenotypic variation in total cholesterol and LDL-cholesterol, respectively. The 2 allele at codon 360 was associated with marginally reduced plasma LDL-cholesterol (P = 0.09) and increased triglyceride (P = 0.05) levels compared to the 1 allele. To further elucidate the combined effects of the two polymorphism we constructed two-site haplotypes. The haplotype data showed a stronger influence and explained 3.0 and 5.2% of the phenotypic variation in total cholesterol and LDL-cholesterol, respectively. The two uncommon haplotypes, T1 and A2, were associated with 24.2 and 23.5 mg/dl lower total cholesterol and 22.5 and 42.0 mg/dl lower LDL cholesterol levels, respectively. The accentuated effect of apo A4 polymorphisms on non-fasting plasma cholesterol suggest that apo A-IV may play an important role in regulating the postprandial metabolism of lipoproteins. PMID- 9199280 TI - Distribution of apolipoprotein E between apo B- and non apo B-containing lipoproteins according to apo E phenotype. AB - Apolipoprotein E (apo E) is a component of all the classes of lipoproteins and can be distributed among apo B- (LpB) and non apo B-containing lipoproteins (Lp non-B). Using a new electroimmunoassay kit, plasma apo E, apo E in Lp-non-B (apo E-Lp-non-B) and apo E in LpB (apo E-LpB) levels were measured in healthy control subjects (n=481) from 3 centers participating in the ECTIM study (Etude Cas Temoins sur l'Infarctus du Myocarde), a population-based study on myocardial infarction. The distribution of apo E among lipoproteins was analyzed according to the apo E phenotype after adjustment for center, body mass index, tobacco use, alcohol consumption and triglycerides. Apo E was higher (average excess: + 0.32; P < 0.0001) and lower (average excess: -0.12; P < 0.0001) in subjects carrying the allele epsilon2 and the allele epsilon4 respectively, than in apo E3/3 subjects. These differences are the consequence of variations in apo E-Lp-non-B which clearly differed between the groups classified according to their apo E phenotype (P < 0.0001). The average excess of apo E Lp non-B compared to apo E3/3 subjects was + 0.43 (P < 0.0001) and -0.22 (P < 0.0001) for the epsilon2 and epsilon4 alleles respectively. Apo E-LpB was lower in subjects carrying the epsilon2 allele (P < 0.02) while the presence of the epsilon4 allele did not modify this parameter. The proportion of apo E within HDL was clearly higher and lower in subjects carrying apo E2 and apo E4 respectively than in apo E3/3 subjects. Although triglyceride levels were dependent on the apo E phenotype, the adjustment of the proportion of apo E in HDL for triglycerides hardly modified the results. For the first time, these results, using direct measurements on a large number of subjects, confirm the greater preference of apo E4 over apo E2 for LpB and vice versa for Lp-non-B. They also show a greater affinity of apo E2 for HDL compared to apo E3. This high affinity of apo E2 for HDL could be due to the formation of the apo E-A-II complex. These results indicate that apo E phenotype modulates the distribution of apo E among lipoproteins and suggest differences in lipoprotein metabolism between apo E2, apo E3 and apo E4. PMID- 9199281 TI - Simvastatin reduces forearm vascular responsiveness to norepinephrine. PMID- 9199282 TI - The branchpoint residue is recognized during commitment complex formation before being bulged out of the U2 snRNA-pre-mRNA duplex. AB - We have analyzed the mechanism of branchpoint nucleotide selection during the first step of pre-mRNA splicing. It has previously been proposed that the branchpoint is selected as an adenosine residue bulged out of an RNA helix formed by the U2 snRNA-pre-mRNA base pairing. Although compatible with this bulge hypothesis, available data from both yeast and mammalian systems did not rule out alternative structures for the branch nucleotide. Mutating the residue preceding the branchpoint nucleotide in our reporter construct conferred a splicing defect that was suppressed in vivo by the complementary U2 snRNA mutants. In contrast, substitutions on the 3' side of the branchpoint could be suppressed by complementary U2 snRNA mutants only in a weakened intron context. To test why the identity of the branch nucleotide was important for its selection, we analyzed the effect of substitutions at this position on spliceosome assembly. We observed that these mutations block the formation of one of the two commitment complexes. Our results demonstrate that yeast branchpoint selection occurs in multiple steps. The nature of the branch residue is recognized, in the absence of U2 snRNA, during commitment complex formation. Then, base pairing with U2 snRNA constrains this residue into a bulge conformation. PMID- 9199283 TI - Coordinate transcription and V(D)J recombination of the kappa immunoglobulin light-chain locus: NF-kappaB-dependent and -independent pathways of activation. AB - To further elucidate the potential role of mitogens and cytokines in regulation of the kappa immunoglobulin light-chain locus, we have characterized the activation of transcription factor binding, kappa germ line transcription, DNase I hypersensitivity, and Vkappa-to-Jkappa recombination upon induction of model pre-B-cell lines. We find that both lipopolysaccharide (LPS) and gamma interferon (IFN-gamma) are capable of activating germ line transcription, DNase I hypersensitivity, and recombination of the kappa locus. We also find that transforming growth factor beta is capable of completely inhibiting LPS activation of transcription and recombination but has no apparent effect on activation of transcription factor binding, including activation of NF-kappaB. To address the functional role of NF-kappaB in LPS and IFN-gamma induction of these events, we blocked the nuclear translocation of NF-kappaB by overexpression of a dominant negative mutant of IkappaB-alpha (IkappaB deltaN). Overexpression of the IkappaB deltaN protein results in an inhibition of LPS but not IFN-gamma activation of germ line transcription, DNase I hypersensitivity, and Vkappa-to Jkappa recombination. Our results demonstrate that activation of NF-kappaB is necessary but not sufficient for LPS activation of transcription and recombination at kappa. These results also suggest that NF-kappaB is not required for IFN-gamma activation of transcription or recombination. These results are important in establishing that there are multiple independent pathways of activation of both transcription and recombination. PMID- 9199284 TI - Transcriptional synergy between LIM-homeodomain proteins and basic helix-loop helix proteins: the LIM2 domain determines specificity. AB - LIM-homeodomain proteins direct cellular differentiation by activating transcription of cell-type-specific genes, but this activation requires cooperation with other nuclear factors. The LIM-homeodomain protein Lmx1 cooperates with the basic helix-loop-helix (bHLH) protein E47/Pan-1 to activate the insulin promoter in transfected fibroblasts. In this study, we show that two proteins originally called Lmx1 are the closely related products of two distinct vertebrate genes, Lmx1.1 and Lmx1.2. We have used yeast genetic systems to delineate the functional domains of the Lmx1 proteins and to characterize the physical interactions between Lmx1 proteins and E47/Pan-1 that produce synergistic transcriptional activation. The LIM domains of the Lmx1 proteins, and particularly the second LIM domain, mediate both specific physical interactions and transcriptional synergy with E47/Pan-1. The LIM domains of the LIM homeodomain protein Isl-1, which cannot mediate transcriptional synergy with E47/Pan-1, do not interact with E47/Pan-1. In vitro studies demonstrate that the Lmx1.1 LIM2 domain interacts specifically with the bHLH domain of E47/Pan-1. These studies provide the basis for a model of the assembly of LIM-homeodomain containing complexes on DNA elements that direct cell-type-restricted transcription in differentiated tissues. PMID- 9199285 TI - Transactivation domains facilitate promoter occupancy for the dioxin-inducible CYP1A1 gene in vivo. AB - We have studied the transcriptional regulation of the dioxin-inducible mouse CYP1A1 gene in its native chromosomal setting. We analyzed the ability of aromatic hydrocarbon receptor (AhR) mutants and AhR chimeras to restore dioxin responsiveness to the CYP1A1 gene in AhR-defective mouse hepatoma cells. Our data reveal that transactivation domains in AhR's C-terminal half mediate occupancy of the nuclear factor 1 site and TATA box for the CYP1A1 promoter in vivo. Transactivation domains of VP16 and AhR nuclear translocator, but not Sp1, can substitute for AhR's C-terminal half in facilitating protein binding at the promoter. Our data also reveal an apparent linear relationship between promoter occupancy and CYP1A1 gene expression in chromatin. These findings provide new insights into the in vivo mechanism of transcriptional activation for an interesting mammalian gene. PMID- 9199286 TI - Mkh1, a MEK kinase required for cell wall integrity and proper response to osmotic and temperature stress in Schizosaccharomyces pombe. AB - We have identified a Schizosaccharomyces pombe gene, mkh1, that encodes a MEK kinase (MEKK) homolog. The coding region of mkh1 is contained within a single exon encoding a 1,116-amino-acid protein. The putative catalytic domain of Mkh1 is 54% identical to the catalytic domain of S. cerevisiae Bck1, the most closely related protein. Deletion of mkh1 did not significantly affect cell growth or division under standard conditions. However, mkh1delta cell growth was inhibited by high KCl or NaCl concentrations. mkh1delta cells required a longer time to reenter the cell cycle after prolonged stationary-phase arrest. Also, mkh1delta cells exhibited a round cell shape, while overexpression of Mkh1 resulted in an elongated cell shape. mkh1delta cells exhibited a more dramatic phenotype when grown in nutrient-limiting conditions at high temperature or in hyperosmotic medium. In such conditions, completion of cytokinesis was inhibited, resulting in the growth of pseudohyphal filaments with multiple septa and nuclei. Also, mkh1delta cells were hypersensitive to beta-glucanase treatment. Together these results suggest that Mkh1 regulates cell morphology, cell wall integrity, salt resistance, cell cycle reentry from stationary-phase arrest, and filamentous growth in response to stress. These phenotypes are essentially identical to those exhibited by cells lacking Pmk1/Spm1, a recently identified mitogen-activated protein kinase. Our evidence suggests that Pmk1/Spm1 acts downstream from Mkh1 in a common pathway. Our results also suggest that Mkh1 and Pck2 act independently to maintain cell wall integrity, cell morphology, and salt resistance but act in opposition to regulate filamentous growth. PMID- 9199287 TI - Topoisomerase function during replication-independent chromatin assembly in yeast. AB - DNA topoisomerases I and II are the two major nuclear enzymes capable of relieving torsional strain in DNA. Of these enzymes, topoisomerase I plays the dominant role in relieving torsional strain during chromatin assembly in cell extracts from oocytes, eggs, and early embryos. We tested if the topoisomerases are used differentially during chromatin assembly in Saccharomyces cerevisiae by a combined biochemical and pharmacological approach. As measured by plasmid supercoiling, nucleosome deposition is severely impaired in assembly extracts from a yeast mutant with no topoisomerase I and a temperature-sensitive form of topoisomerase II (strain top1-top2). Expression of wild-type topoisomerase II in strain top1-top2 fully restored assembly-driven supercoiling, and assembly was equally efficient in extracts from strains expressing either topoisomerase I or II alone. Supercoiling in top1-top2 extract was rescued by adding back either purified topoisomerase I or II. Using the topoisomerase II poison VP-16, we show that topoisomerase II activity during chromatin assembly is the same in the presence and absence of topoisomerase I. We conclude that both topoisomerases I and II can provide the DNA relaxation activity required for efficient chromatin assembly in mitotically cycling yeast cells. PMID- 9199288 TI - Combinatorial determinants of tissue-specific transcription in B cells and macrophages. AB - A tripartite domain of the immunoglobulin mu heavy-chain gene enhancer that activates transcription in B cells contains binding sites for PU.1, Ets-1, and a leucine zipper-containing basic helix-loop-helix factor. Because PU.1 is expressed only in B cells and macrophages, we tested the activity of a minimal mu enhancer fragment in macrophages by transient transfections. The minimal mu enhancer activated transcription in macrophages, and the activity was dependent on all three sites. Analysis of mutated enhancers, in which spacing and orientation of the ETS protein binding sites had been changed, suggested that the mechanisms of enhancer activation were different in B cells and macrophages. Thus, ETS protein binding sites may be combined in different ways to generate tissue-specific transcription activators. Despite the activity of the minimal enhancer in macrophages, a larger mu enhancer fragment was inactive in these cells. We propose that formation of the nucleoprotein complex that is formed on the minimal enhancer in macrophages cannot be helped by the neighboring muE elements that are essential for activity of the monomeric enhancer. PMID- 9199289 TI - Analysis of a meiosis-specific URS1 site: sequence requirements and involvement of replication protein A. AB - URS1 is a transcriptional repressor site found in the promoters of a wide variety of yeast genes that are induced under stress conditions. In the context of meiotic promoters, URS1 sites act as repressor sequences during mitosis and function as activator sites during meiosis. We have investigated the sequence requirements of the URS1 site of the meiosis-specific HOP1 gene (URS1H) and have found differences compared with a URS1 site from a nonmeiotic gene. We have also observed that the sequence specificity for meiotic activation at this site differs from that for mitotic repression. Base pairs flanking the conserved core sequence enhance meiotic induction but are not required for mitotic repression of HOP1. Electrophoretic mobility shift assays of mitotic and meiotic cell extracts show a complex pattern of DNA-protein complexes, suggesting that several different protein factors bind specifically to the site. We have determined that one of the complexes of URS1H is formed by replication protein A (RPA). Although RPA binds to the double-stranded URS1H site in vitro, it has much higher affinity for single-stranded than for double-stranded URS1H, and one-hybrid assays suggest that RPA does not bind to this site at detectable levels in vivo. In addition, conditional-lethal mutations in RPA were found to have no effect on URS1H mediated repression. These results suggest that although RPA binds to URS1H in vitro, it does not appear to have a functional role in transcriptional repression through this site in vivo. PMID- 9199290 TI - Signaling through mitogen-activated protein kinase and Rac/Rho does not duplicate the effects of activated Ras on skeletal myogenesis. AB - The ability of basic helix-loop-helix muscle regulatory factors (MRFs), such as MyoD, to convert nonmuscle cells to a myogenic lineage is regulated by numerous growth factor and oncoprotein signaling pathways. Previous studies have shown that H-Ras 12V inhibits differentiation to a skeletal muscle lineage by disrupting MRF function via a mechanism that is independent of the dimerization, DNA binding, and inherent transcriptional activation properties of the proteins. To investigate the intracellular signaling pathway(s) that mediates the inhibition of MRF-induced myogenesis by oncogenic Ras, we tested two transformation-defective H-Ras 12V effector domain variants for their ability to alter terminal differentiation. H-Ras 12V,35S retains the ability to activate the Raf/MEK/mitogen-activated protein (MAP) kinase cascade, whereas H-Ras 12V,40C is unable to interact directly with Raf-1 yet still influences other signaling intermediates, including Rac and Rho. Expression of each H-Ras 12V variant in C3H10T1/2 cells abrogates MyoD-induced activation of the complete myogenic program, suggesting that MAP kinase-dependent and -independent Ras signaling pathways individually block myogenesis in this model system. However, additional studies with constitutively activated Rac1 and RhoA proteins revealed no negative effects on MyoD-induced myogenesis. Similarly, treatment of Ras-inhibited myoblasts with the MEK1 inhibitor PD98059 revealed that elevated MAP kinase activity is not a significant contributor to the H-Ras 12V effect. These data suggest that an additional Ras pathway, distinct from the well-characterized MAP kinase and Rac/Rho pathways known to be important for the transforming function of activated Ras, is primarily responsible for the inhibition of myogenesis by H Ras 12V. PMID- 9199291 TI - Differential regulation of the mitogen-activated protein and stress-activated protein kinase cascades by adrenergic agonists in quiescent and regenerating adult rat hepatocytes. AB - To study the mechanisms by which catecholamines regulate hepatocyte proliferation after partial hepatectomy (PHX), hepatocytes were isolated from adult male rats 24 h after sham operation or two-thirds PHX and treated with catecholamines and other agonists. In freshly isolated sham cells, p42 mitogen-activated protein (MAP) kinase activity was stimulated by the alpha1-adrenergic agonist phenylephrine (PHE). Activation of p42 MAP kinase by growth factors was blunted by pretreatment of sham hepatocytes with glucagon but not by that with the beta2 adrenergic agonist isoproterenol (ISO). In PHX cells, the ability of PHE to activate p42 MAP kinase was dramatically reduced, whereas ISO became competent to inhibit p42 MAP kinase activation. PHE treatment of sham but not PHX and ISO treatment of PHX but not sham hepatocytes also activated the stress-activated protein (SAP) kinases p46/54 SAP kinase and p38 SAP kinase. These data demonstrate that an alpha1- to beta2-adrenergic receptor switch occurs upon PHX and results in an increase in SAP kinase versus MAP kinase signaling by catecholamines. In primary cultures of hepatocytes, ISO treatment of PHX but not sham cells inhibited [3H]thymidine incorporation. In contrast, PHE treatment of sham but not PHX cells stimulated [3H]thymidine incorporation, which was reduced by approximately 25 and approximately 95% with specific inhibitors of p42 MAP kinase and p38 SAP kinase function, respectively. Inhibition of the p38 SAP kinase also dramatically reduced basal [3H]thymidine incorporation. These data suggest that p38 SAP kinase plays a permissive role in liver regeneration. Alterations in the abilities of catecholamines to modulate the activities of protein kinase A and the MAP and SAP kinase pathways may represent one physiological mechanism by which these agonists can regulate hepatocyte proliferation after PHX. PMID- 9199292 TI - p130 and p107 use a conserved domain to inhibit cellular cyclin-dependent kinase activity. AB - The pRB-related proteins p107 and p130 are thought to suppress growth in part through their associations with two important cell cycle kinases, cyclin A-cdk2 and cyclin E-cdk2, and transcription factor E2F. Although each protein plays a critical role in cell proliferation, the functional consequences of the association among growth suppressor, cyclin-dependent kinase, and transcription factor have remained elusive. In an attempt to understand the biochemical properties of such complexes, we reconstituted each of the p130-cyclin-cdk2 and p107-cyclin-cdk2 complexes found in vivo with purified, recombinant proteins. Strikingly, stoichiometric association of p107 or p130 with either cyclin E-cdk2 or cyclin A-cdk2 negated the activities of these kinases. The results of our experiments suggest that inhibition does not result from substrate competition or loss of cdk2 activation. Kinase inhibitory activity was dependent upon an amino terminal region of p107 that is highly conserved with p130. Further, a role for this amino-terminal region in growth suppression was uncovered by using p107 mutants unable to bind E2F. To determine whether cellular complexes might display similar regulatory properties, we purified p130-cyclin A-cdk2 complexes from human cells and found that such complexes exist in two forms, one that contains E2F-4-DP-1 and one that lacks the heterodimer. These endogenous complexes behaved like the in vitro-reconstituted complexes, exhibiting low levels of associated kinase activity that could be significantly augmented by dissociation of p130. The results of these experiments suggest a mechanism whereby p130 and p107 suppress growth by inhibiting important cell cycle kinases. PMID- 9199293 TI - Prp31p promotes the association of the U4/U6 x U5 tri-snRNP with prespliceosomes to form spliceosomes in Saccharomyces cerevisiae. AB - The PRP31 gene encodes a factor essential for the splicing of pre-mRNA in Saccharomyces cerevisiae. Cell extracts derived from a prp31-1 strain fail to form mature spliceosomes upon heat inactivation, although commitment complexes and prespliceosome complexes are detected under these conditions. Coimmunoprecipitation experiments indicate that Prp31p is associated both with the U4/U6 x U5 tri-snRNP and, independently, with the prespliceosome prior to assembly of the tri-snRNP into the splicing complex. Nondenaturing gel electrophoresis and glycerol gradient analyses demonstrate that while Prp31p may play a role in maintaining the assembly or stability of tri-snRNPs, functional protein is not essential for the formation of U4/U6 or U4/U6 x U5 snRNPs. These results suggest that Prp31p is involved in recruiting the U4/U6 x U5 tri-snRNP to prespliceosome complexes or in stabilizing these interactions. PMID- 9199294 TI - Sequence-specific epigenetic effects of the maternal somatic genome on developmental rearrangements of the zygotic genome in Paramecium primaurelia. AB - In ciliates, the germ line genome is extensively rearranged during the development of the somatic macronucleus from a mitotic product of the zygotic nucleus. Germ line chromosomes are fragmented in specific regions, and a large number of internal sequence elements are eliminated. It was previously shown that transformation of the vegetative macronucleus of Paramecium primaurelia with a plasmid containing a subtelomeric surface antigen gene can affect the processing of the homologous germ line genomic region during development of a new macronucleus in sexual progeny of transformed clones. The gene and telomere proximal flanking sequences are deleted from the new macronuclear genome, although the germ line genome remains wild type. Here we show that plasmids containing nonoverlapping segments of the same genomic region are able to induce similar terminal deletions; the locations of deletion end points depend on the particular sequence used. Transformation of the maternal macronucleus with a sequence internal to a macronuclear chromosome also causes the occurrence of internal deletions between short direct repeats composed of alternating thymines and adenines. The epigenetic influence of maternal macronuclear sequences on developmental rearrangements of the zygotic genome thus appears to be both sequence specific and general, suggesting that this trans-nucleus effect is mediated by pairing of homologous sequences. PMID- 9199295 TI - CREB controls LAP/C/EBP beta transcription. AB - LAP/C/EBP beta is a member of the C/EBP family of transcription factors and is involved in hepatocyte-specific gene expression. Recently we showed that, besides its posttranscriptional regulation, LAP/C/EBP beta mRNA is modulated during liver regeneration. Therefore, in this study we investigated mechanisms which control LAP/C/EBP beta gene transcription. Deletion analysis of the 5'-flanking region, located upstream of the start site of transcription in the LAP/C/EBP beta gene, demonstrated that a small region in close proximity to the TATA box is important in maintaining a high level of transcription of the luciferase reporter gene constructs. In gel shift experiments two sites were identified which are important for specific complex formation within this region. Further analysis by cross-linking, super shift, and competition experiments was performed with liver cell nuclear extracts, hepatoma cell nuclear extracts, or recombinant CREB protein. These experiments conclusively demonstrated that CREB binds to both sites in the LAP/C/EBP beta promoter with an affinity similar to that with the CREB consensus sequence. Transfection experiments with promoter constructs where the CREB sites were mutated showed that these sites are important to maintain both basal promoter activity and LAP/C/EBP beta inducibility through CREB. Northern blot analysis and runoff transcription assays demonstrated that the protein kinase A pathway not only stimulated the activity of the luciferase reporter construct but also the transcription of the endogenous LAP/C/EBP beta gene in different cell types. Western blot analysis of rat liver cell nuclear extracts and runoff transcription assays of rat liver cell nuclei after two thirds hepatectomy showed a functional link between the induction of CREB phosphorylation and LAP/C/EBP beta mRNA transcription during liver regeneration. These results demonstrate that the two CREB sites are important to control LAP/C/EBP beta transcription in vivo. As several pathways control CREB phosphorylation, our results provide evidence for the transcriptional regulation of LAP/C/EBP beta via CREB under different physiological conditions. PMID- 9199296 TI - Fine-resolution analysis of products of intrachromosomal homeologous recombination in mammalian cells. AB - Mouse Ltk- cell lines that contained a herpes simplex virus type 1 (HSV-1) thymidine kinase (tk) gene with a 16-bp insertion mutation linked to either a defective HSV-2 tk gene or a hybrid tk sequence comprised of HSV-1 and HSV-2 tk sequences were constructed. HSV-1 and HSV-2 tk genes have 81% nucleotide identity and hence are homeologous. Correction of the insertion mutant HSV-1 tk gene via recombination with the hybrid tk sequence required an exchange between homeologous tk sequences, although recombination could initiate within a region of significant sequence identity. Seven cell lines containing linked HSV-1 and HSV-1-HSV-2 hybrid tk sequences gave rise to tk+ segregants at an average rate of 10(-8) events per cell division. DNA sequencing revealed that each recombinant from these lines displayed an apparent gene conversion which involved an accurate transfer of an uninterrupted block of information between homeologous tk sequences. Conversion tract lengths ranged from 35 to >330 bp. In contrast, cell lines containing linked HSV-1 and HSV-2 tk sequences with no significant stretches of sequence identity had an overall rate of homeologous recombination of <10(-9). One such cell line produced homeologous recombinants at a rate of 10( 8). Strikingly, all homeologous recombinants from this latter cell line were due to crossovers between the HSV-1 and HSV-2 tk genes. Our results, which provide the first detailed analysis of homeologous recombination within a mammalian genome, suggest that rearrangements in mammalian genomes are regulated by the degree of sequence divergence located at the site of recombination initiation. PMID- 9199297 TI - Constitutive activation of NF-kappaB during progression of breast cancer to hormone-independent growth. AB - Breast cancers often progress from a hormone-dependent, nonmetastatic, antiestrogen-sensitive phenotype to a hormone-independent, antiestrogen- and chemotherapy-resistant phenotype with highly invasive and metastatic growth properties. This progression is usually accompanied by altered function of the estrogen receptor (ER) or outgrowth of ER-negative cancer cells. To understand the molecular mechanisms responsible for metastatic growth of ER-negative breast cancers, the activities of the transcription factor NF-kappaB (which modulates the expression of genes involved in cell proliferation, differentiation, apoptosis, and metastasis) were compared in ER-positive (MCF-7 and T47-D) and ER negative (MDA-MB-231 and MDA-MB-435) human breast cancer cell lines. NF-kappaB, which is usually maintained in an inactive state by protein-protein interaction with inhibitor IkappaBs, was found to be constitutively active in ER-negative breast cancer cell lines. Constitutive DNA binding of NF-kappaB was also observed with extracts from ER-negative, poorly differentiated primary breast tumors. Progression of the rat mammary carcinoma cell line RM22-F5 from an ER-positive, nonmalignant phenotype (E phenotype) to an ER-negative, malignant phenotype (F phenotype) was also accompanied by constitutive activation of NF-kappaB. Analysis of individual subunits of NF-kappaB revealed that all ER-negative cell lines, including RM22-F5 cells of F phenotype, contain a unique 37-kDa protein which is antigenically related to the RelA subunit. Cell-type-specific differences in IkappaB alpha, -beta, and -gamma were also observed. In transient-transfection experiments, constitutive activity of an NF-kappaB-dependent promoter was observed in MDA-MB-231 and RM22-F5 cells of F phenotype, and this activity was efficiently repressed by cotransfected ER. Since ER inhibits the constitutive as well as inducible activation function of NF-kappaB in a dose-dependent manner, we propose that breast cancers that lack functional ER overexpress NF-kappaB regulated genes. Furthermore, since recent data indicate that NF-kappaB protects cells from tumor necrosis factor alpha-, ionizing radiation-, and chemotherapeutic agent daunorubicin-mediated apoptosis, our results provide an explanation for chemotherapeutic resistance in ER-negative breast cancers. PMID- 9199298 TI - Met31p and Met32p, two related zinc finger proteins, are involved in transcriptional regulation of yeast sulfur amino acid metabolism. AB - Sulfur amino acid metabolism in Saccharomyces cerevisiae is regulated by the level of intracellular S-adenosylmethionine (AdoMet). Two cis-acting elements have been previously identified within the 5' upstream regions of the structural genes of the sulfur network. The first contains the CACGTG motif and is the target of the transcription activation complex Cbflp-Met4p-Met28p. We report here the identification of two new factors, Met31p and Met32p, that recognize the second cis-acting element. Met31p was isolated through the use of the one-hybrid method, while Met32p was identified during the analysis of the yeast methionine transport system. Met31p and Met32p are highly related zinc finger-containing proteins. Both LexA-Met31p and LexA-Met32p fusion proteins activate the transcription of a LexAop-containing promoter in a Met4p-dependent manner. Northern blot analyses of cells that do not express either Met31p and/or Met32p suggest that the function of the two proteins during the transcriptional regulation of the sulfur network varies from one gene to the other. While the expression of both the MET3 and MET14 genes was shown to strictly depend upon the presence of either Met31p or Met32p, the transcription of the MET25 gene is constitutive in cells lacking both Met31p and Met32p. These results therefore emphasise the diversity of the mechanisms allowing regulation of the expression of the methionine biosynthetic genes. PMID- 9199299 TI - Intra- and intermolecular cooperative binding of high-mobility-group protein I(Y) to the beta-interferon promoter. AB - The mammalian high-mobility-group protein I(Y) [HMG I(Y)], while not a typical transcriptional activator, is required for the expression of many eukaryotic genes. HMG I(Y) appears to recruit and stabilize complexes of transcriptional activators through protein-DNA and protein-protein interactions. The protein binds to the minor groove of DNA via three short basic repeats, preferring tracts of adenines and thymines arranged on the same face of the DNA helix. However, the mode by which these three basic repeats function together to recognize HMG I(Y) binding sites has remained unclear. Here, using deletion mutants of HMG I(Y), DNase I footprinting, methylation interference, and in vivo transcriptional assays, we have characterized the binding of HMG I(Y) to the model beta interferon enhancer. We show that two molecules of HMG I(Y) bind to the enhancer in a highly cooperative fashion, each molecule using a distinct pair of basic repeats to recognize the tandem AT-rich regions of the binding sites. We have also characterized the function of each basic repeat, showing that only the central repeat accounts for specific DNA binding and that the presence of a second repeat bound to an adjacent AT-rich region results in intramolecular cooperativity in binding. Surprisingly, the carboxyl-terminal acidic tail of HMG I(Y) is also important for specific binding in the context of the full-length protein. Our results present a detailed examination of HMG I(Y) binding in an important biological context, which can be extended not only to HMG I(Y) binding in other systems but also to the binding mode of many other proteins containing homologous basic repeats, which have been conserved from bacteria to humans. PMID- 9199300 TI - Comparison of the transactivation domains of Stat5 and Stat6 in lymphoid cells and mammary epithelial cells. AB - Stat (signal transducers and activators of transcription) and Jak (Janus kinases) proteins are central components in the signal transduction events in hematopoietic and epithelial cells. They are rapidly activated by various cytokines, hormones, and growth factors. Upon ligand binding and cytokine receptor dimerization, Stat proteins are phosphorylated on tyrosine residues by Jak kinases. Activated Stat proteins form homo- or heterodimers, translocate to the nucleus, and induce transcription from responsive genes. Stat5 and Stat6 are transcription factors active in mammary epithelial cells and immune cells. Prolactin activates Stat5, and interleukin-4 (IL-4) activates Stat6. Both cytokines are able to stimulate cell proliferation, differentiation, and survival. We investigated the transactivation potential of Stat6 and found that it is not restricted to lymphocytes. IL-4-dependent activation of Stat6 was also observed in HC11 mammary epithelial cells. In these cells, Stat6 activation led to the induction of the beta-casein gene promoter. The induction of this promoter was confirmed in COS7 cells. The glucocorticoid receptor was able to further enhance IL-4-induced gene transcription through the action of Stat6. Deletion analysis of the carboxyl-terminal region of Stat6 and recombination of this region with a heterologous DNA binding domain allowed the delimitation and characterization of the transactivation domain of Stat6. The potencies of the transactivation domains of Stat5, Stat6, and viral protein VP16 were compared. Stat6 had a transactivation domain which was about 10-fold stronger than that of Stat5. In pre-B cells (Ba/F3), the transactivation domain of Stat6 was IL-4 regulated, independently from its DNA binding function. PMID- 9199301 TI - Expression of var genes located within polymorphic subtelomeric domains of Plasmodium falciparum chromosomes. AB - Plasmodium falciparum var genes encode a diverse family of proteins, located on the surfaces of infected erythrocytes, which are implicated in the pathology of human malaria through antigenic variation and adhesion of infected erythrocytes to the microvasculature. We have constructed a complete representative telomere to-telomere yeast artificial chromosome (YAC) contig map of the P. falciparum chromosome 8 for studies on the chromosomal organization, distribution, and expression of var genes. Three var gene loci were identified on chromosome 8, two of which map close to the telomeres at either end of the chromosome. Analysis of the previously described chromosome 2 contig map and random P. falciparum telomeric YAC clones revealed that most, if not all, 14 P. falciparum chromosomes contain var genes in a subtelomeric location. Mapping the chromosomal location of var genes expressed in a long-term culture of the P. falciparum isolate Dd2 revealed that four of the five different expressed var genes identified map within subtelomeric locations. Expression of var genes from a chromosomal domain known for frequent rearrangements has important implications for the mechanism of var gene switching and the generation of novel antigenic and adhesive phenotypes. PMID- 9199302 TI - A circadian enhancer mediates PER-dependent mRNA cycling in Drosophila melanogaster. AB - Genes expressed under circadian-clock control are found in organisms ranging from prokaryotes to humans. In Drosophila melanogaster, the period (per) gene, which is required for clock function, is transcribed in a circadian manner. We have identified a circadian transcriptional enhancer within a 69-bp DNA fragment upstream of the per gene. This enhancer drives high-amplitude mRNA cycling under light-dark-cycling or constant-dark conditions, and this activity is per protein (PER) dependent. An E-box sequence within this 69-bp fragment is necessary for high-level expression, but not for rhythmic expression, indicating that PER mediates circadian transcription through other sequences in this fragment. PMID- 9199303 TI - Yeast Pab1 interacts with Rna15 and participates in the control of the poly(A) tail length in vitro. AB - In Saccharomyces cerevisiae, the single poly(A) binding protein, Pab1, is the major ribonucleoprotein associated with the poly(A) tails of mRNAs in both the nucleus and the cytoplasm. We found that Pab1 interacts with Rna15 in two-hybrid assays and in coimmunoprecipitation experiments. Overexpression of PAB1 partially but specifically suppressed the rna15-2 mutation in vivo. RNA15 codes for a component of the cleavage and polyadenylation factor CF I, one of the four factors needed for pre-mRNA 3'-end processing. We show that Pab1 and CF I copurify in anion-exchange chromatography. These data suggest that Pab1 is physically associated with CF I. Extracts from a thermosensitive pab1 mutant and from a wild-type strain immunoneutralized for Pab1 showed normal cleavage activity but a large increase in poly(A) tail length. A normal tail length was restored by adding recombinant Pab1 to the mutant extract. The longer poly(A) tails were not due to an inhibition of exonuclease activities. Pab1 has previously been implicated in the regulation of translation initiation and in cytoplasmic mRNA stability. Our data indicate that Pab1 is also a part of the 3' end RNA-processing complex and thus participates in the control of the poly(A) tail lengths during the polyadenylation reaction. PMID- 9199304 TI - The sequence of the 5' end of the U8 small nucleolar RNA is critical for 5.8S and 28S rRNA maturation. AB - Ribosome biogenesis in eucaryotes involves many small nucleolar ribonucleoprotein particles (snoRNP), a few of which are essential for processing pre-rRNA. Previously, U8 snoRNA was shown to play a critical role in pre-rRNA processing, being essential for accumulation of mature 28S and 5.8S rRNAs. Here, evidence which identifies a functional site of interaction on the U8 RNA is presented. RNAs with mutations, insertions, or deletions within the 5'-most 15 nucleotides of U8 do not function in pre-rRNA processing. In vivo competitions in Xenopus oocytes with 2'O-methyl oligoribonucleotides have confirmed this region as a functional site of a base-pairing interaction. Cross-species hybrid molecules of U8 RNA show that this region of the U8 snoRNP is necessary for processing of pre rRNA but not sufficient to direct efficient cleavage of the pre-rRNA substrate; the structure or proteins comprising, or recruited by, the U8 snoRNP modulate the efficiency of cleavage. Intriguingly, these 15 nucleotides have the potential to base pair with the 5' end of 28S rRNA in a region where, in the mature ribosome, the 5' end of 28S interacts with the 3' end of 5.8S. The 28S-5.8S interaction is evolutionarily conserved and critical for pre-rRNA processing in Xenopus laevis. Taken together these data strongly suggest that the 5' end of U8 RNA has the potential to bind pre-rRNA and in so doing, may regulate or alter the pre-rRNA folding pathway. The rest of the U8 particle may then facilitate cleavage or recruitment of other factors which are essential for pre-rRNA processing. PMID- 9199305 TI - The immediate-early gene product Egr-1 regulates the human interleukin-2 receptor beta-chain promoter through noncanonical Egr and Sp1 binding sites. AB - The interleukin-2 IL-2 receptor beta-chain (IL-2Rbeta) is an essential component of the receptors for IL-2 and IL-15. Although IL-2Rbeta is constitutively expressed by lymphocytes, its expression can be further induced by a number of stimuli, including phorbol 12-myristate 13-acetate (PMA). We have now characterized factors that bind to an enhancer region located between nucleotides -170 and -139 of the human IL-2Rbeta promoter. Both Sp1 and Sp3 bound to the 5' portion of this region, whereas a PMA-inducible factor (PIF) mainly bound to its 3' portion and bound to the Sp binding motifs as well. In Jurkat T cells, induction of PIF DNA binding activity was rapidly induced, required de novo protein synthesis, and was sustained at a high level for at least 23 h. Interestingly, PIF was constitutively activated in human T-cell leukemia virus type 1-transformed MT-2 cells. In this paper, we demonstrate that PIF is Egr-1 based on its recognition by anti-Egr-1 antisera in gel mobility shift assays, even though the IL-2Rbeta DNA binding motif differed substantially from the canonical Egr-1 binding site. In addition, Egr-1 bound to the Sp binding site. In Jurkat cells, both sites were required for maximal IL-2Rbeta promoter activity, and in HeLaS3 cells, transfection of Egr-1 could drive activity of a reporter construct containing both sites. Moreover, Sp1 and Egr-1 could form a complex with kinetics that correlated with the production of Egr-1 in Jurkat cells upon PMA stimulation. Thus, Sp1 and Egr-1 physically and functionally cooperate to mediate maximal IL-2Rbeta promoter activity. PMID- 9199306 TI - Multiple mechanisms of transcriptional repression by YY1. AB - The four C-terminal GLI-Kruppel type zinc fingers of YY1 have been identified as a transcriptional repression domain. Previous reports have proposed DNA-bending and activator-quenching mechanisms for this zinc finger-mediated repression. In addition, previous work indicated that p300 and CBP might be involved in YY1 mediated repression. We have analyzed these possible models for the zinc finger mediated repression. The role of each zinc finger in the repression and DNA binding functions was determined by using a structure-and-function approach. We show that zinc finger 2 of YY1 plays a central role in both DNA binding and transcriptional repression. However, a survey of a panel of YY1 mutants indicates that these two functions can be separated, which argues against the DNA-bending model for repression. We show that the physical interaction between YY1 and p300, a coactivator for CREB, is not sufficient for repression of CREB-mediated transcription. Our studies indicate that YY1 functions as an activator-specific repressor. Repression of CTF-1-directed transcription may be accomplished through direct physical interaction between YY1 and this activator. In contrast, physical interaction is not necessary for YY1 to repress Sp1- and CREB-mediated transcription. Rather, the repression likely reflects an ability of YY1 to interfere with communication between these activators and their targets within the general transcription machinery. Taken together, our results suggest that YY1 employs multiple mechanisms to achieve activator-specific repression. PMID- 9199307 TI - Recombination signal sequence binding protein Jkappa is constitutively bound to the NF-kappaB site of the interleukin-6 promoter and acts as a negative regulatory factor. AB - Analysis by electrophoretic mobility shift assays (EMSA) of the different proteins associated with the kappaB sequence of the interleukin-6 (IL-6) promoter (IL6-kappaB) allowed us to detect a specific complex formed with the recombination signal sequence binding protein Jkappa (RBP-Jkappa). Single-base exchanges within the oligonucleotide sequence defined the critical base pairs involved in the interaction between RBP-Jkappa and the IL6-kappaB motif. Binding analysis suggests that the amount of RBP-Jkappa protein present in the nucleus is severalfold higher than the total amount of inducible NF-kappaB complexes but that the latter bind DNA with a 10-fold-higher affinity. A reporter gene study was performed to determine the functional implication of this binding; we found that the constitutive occupancy of the IL6-kappaB site by the RBP-Jkappa protein was responsible for the low basal levels of IL-6 promoter activity in L929sA fibrosarcoma cells and that RBP-Jkappa partially blocked access of NF-kappaB complexes to the IL-6 promoter. We propose that such a mechanism could be involved in the constitutive repression of the IL-6 gene under normal physiological conditions. PMID- 9199308 TI - Insulin-like growth factor I receptor signaling in transformation by src oncogenes. AB - R- cells, a line of mouse embryo fibroblasts with a targeted disruption of the insulin-like growth factor I (IGF-I) receptor genes, are refractory to transformation by several viral and cellular oncogenes. Using colony formation in soft agar as a measure of full transformation, we report here that R- cells can be transformed by v-src, although they still cannot be transformed by the activated c-src527 (mutation at tyrosine 527 to phenylalanine), which readily transforms mouse embryo cells with a wild-type number of IGF-I receptors (W cells). Although v-src is a more potent inducer of tyrosine phosphorylation than c-src527, the extent of phosphorylation of either insulin receptor substrate 1 or Shc, two of the major substrates of the IGF-I receptor, does not seem sufficiently different to explain the qualitative difference in soft agar growth. v-src, however, is considerably more efficient than c-src527 in its ability to tyrosyl phosphorylate, in R- cells, the focal adhesion kinase, Stat1, and p130cas. These results indicate that v-src, but not c-src527, can bypass the requirement for a functional IGF-I receptor in the full transformation of mouse embryo fibroblasts and suggest that qualitative and quantitative differences between the two oncogenes can be used to identify some of the signals relevant to the mechanism(s) of transformation. PMID- 9199309 TI - Interaction between the small GTPase Ran/Gsp1p and Ntf2p is required for nuclear transport. AB - Bidirectional movement of proteins and RNAs across the nuclear envelope requires Ran, a Ras-like GTPase. A genetic screen of the yeast Saccharomyces cerevisiae was performed to isolate conditional alleles of GSP1, a gene that encodes a homolog of Ran. Two temperature-sensitive alleles, gsp1-1 and gsp1-2, were isolated. The mutations in these two alleles map to regions that are structurally conserved between different members of the Ras family. Each mutant strain exhibits various nuclear transport defects. Both biochemical and genetic experiments indicate a decreased interaction between Ntf2p, a factor which is required for protein import, and the mutant GSP1 gene products. Overexpression of NTF2 can suppress the temperature sensitive phenotype of gsp1-1 and gsp1-2 and partially rescue nuclear transport defects. However, overexpression of a mutant allele of NTF2 with decreased binding to Gsp1p cannot rescue the temperature sensitivity of gsp1-1 and gsp1-2. Taken together, these data demonstrate that the interaction between Gsp1p and Ntf2p is critical for nuclear transport. PMID- 9199310 TI - Nucleotide deletion and P addition in V(D)J recombination: a determinant role of the coding-end sequence. AB - During V(D)J recombination, the coding ends to be joined are extensively modified. Those modifications, termed coding-end processing, consist of removal and addition of various numbers of nucleotides. We previously showed in vivo that coding-end processing is specific for each coding end, suggesting that specific motifs in a coding-end sequence influence nucleotide deletion and P-region formation. In this study, we created a panel of recombination substrates containing actual immunoglobulin and T-cell receptor coding-end sequences and dissected the role of each motif by comparing its processing pattern with those of variants containing minimal nucleotide changes from the original sequence. Our results demonstrate the determinant role of specific sequence motifs on coding end processing and also the importance of the context in which they are found. We show that minimal nucleotide changes in key positions of a coding-end sequence can result in dramatic changes in the processing pattern. We propose that each coding-end sequence dictates a unique hairpin structure, the result of a particular energy conformation between nucleotides organizing the loop and the stem, and that the interplay between this structure and specific sequence motifs influences the frequency and location of nicks which open the coding-end hairpin. These findings indicate that the sequences of the coding ends determine their own processing and have a profound impact on the development of the primary B- and T cell repertoires. PMID- 9199311 TI - Processing of DNA prior to illegitimate recombination in mouse cells. AB - In mammalian cells, the predominant pathway of chromosomal integration of exogenous DNA is random or illegitimate recombination; integration by homologous recombination is infrequent. Homologous recombination is initiated at double strand DNA breaks which have been acted on by single-strand exonuclease. To further characterize the relationship between illegitimate and homologous recombination, we have investigated whether illegitimate recombination is also preceded by exonuclease digestion. Heteroduplex DNAs which included strand specific restriction markers at each of four positions were generated. These DNAs were introduced into mouse embryonic stem cells, and stably transformed clones were isolated and analyzed to determine whether there was any strand bias in the retention of restriction markers with respect to their positions. Some of the mismatches appear to have been resolved by mismatch repair. Very significant strand bias was observed in the retention of restriction markers, and there was polarity of marker retention between adjacent positions. We conclude that DNA is frequently subjected to 5'-->3' exonuclease digestion prior to integration by illegitimate recombination and that the length of DNA removed by exonuclease digestion can be extensive. We also provide evidence which suggests that frequent but less extensive 3'-->5' exonuclease processing also occurs. PMID- 9199313 TI - Molecular architecture of the hsp70 promoter after deletion of the TATA box or the upstream regulation region. AB - GAGA factor, TFIID, and paused polymerase are present on the hsp70 promoter in Drosophila melanogaster prior to transcriptional activation. In order to investigate the interplay between these components, mutant constructs were analyzed after they had been transformed into flies on P elements. One construct lacked the TATA box and the other lacked the upstream regulatory region where GAGA factor binds. Transcription of each mutant during heat shock was at least 50 fold less than that of a normal promoter construct. Before and after heat shock, both mutant promoters were found to adopt a DNase I hypersensitive state that included the region downstream from the transcription start site. High-resolution analysis of the DNase I cutting pattern identified proteins that could be contributing to the hypersensitivity. GAGA factor footprints were clearly evident in the upstream region of the TATA deletion construct, and a partial footprint possibly caused by TFIID was evident on the TATA box of the upstream deletion construct. Permanganate treatment of intact salivary glands was used to further characterize each promoter construct. Paused polymerase and TFIID were readily detected on the normal promoter construct, whereas both deletions exhibited reduced levels of each of these factors. Hence both the TATA box and the upstream region are required to efficiently recruit TFIID and a paused polymerase to the promoter prior to transcriptional activation. In contrast, GAGA factor appears to be capable of binding and establishing a DNase I hypersensitive region in the absence of TFIID and polymerase. Interestingly, purified GAGA factor was found to bind near the transcription start site, and the strength of this interaction was increased by the presence of the upstream region. GAGA factor alone might be capable of establishing an open chromatin structure that encompasses the upstream regulatory region as well as the core promoter region, thus facilitating the binding of TFIID. PMID- 9199312 TI - Purification and characterization of FBI-1, a cellular factor that binds to the human immunodeficiency virus type 1 inducer of short transcripts. AB - The human immunodeficiency virus (HIV-1) promoter directs the synthesis of two classes of RNA molecules, short transcripts and full-length transcripts. The synthesis of short transcripts depends on a bipartite DNA element, the inducer of short transcripts (IST), located in large part downstream of the HIV-1 start site of transcription. IST does not require any viral product for function and is thought to direct the assembly of transcription complexes that are incapable of efficient elongation. Nothing is known, however, about the biochemical mechanisms that mediate IST function. Here, we report the identification and purification of a factor that binds specifically to the IST. This factor, FBI-1, recognizes a large bipartite binding site that coincides with the bipartite IST element. It is constituted at least in part by an 86-kDa polypeptide that can be specifically cross-linked to IST. FBI-1 also binds to promoter and attenuation regions of a number of cellular and viral transcription units that are regulated by a transcription elongation block. This observation, together with the observation that the binding of FBI-1 to IST mutants correlates with the ability of these mutants to direct IST function, suggests that FBI-1 may be involved in the establishment of abortive transcription complexes. PMID- 9199314 TI - Transcription reinitiation rate: a special role for the TATA box. AB - Promoters need to specify both the timing of transcriptional induction and the amount of transcript synthesized. In order to explore each of these effects separately, in vitro assays for the level of active preinitiation complex formation and for the rate of continuous RNA production were done. The effects were found to be influenced differently by different promoter elements. A consensus TATA element had a very strong effect on the rate of continuous RNA production, whereas two types of activators were important primarily in forming active transcription preinitiation complexes. Consensus TATA promoters exhibited high rates of continuous transcription; they assembled active preinitiation transcription complexes slowly but then produced transcripts continuously at an approximately fivefold-higher rate. Initiator-containing TATA-less promoters produced continuous transcripts slowly. Point mutations in the TATA element led to lower levels of transcription by reducing the number of preinitiation complexes and amplifying this reduction by lowering the apparent reinitiation rate. The results allow understanding of the sequence diversity of promoter elements in terms of specifying separate controls over the sensitivity of gene induction and over the strength of the induced promoter. PMID- 9199315 TI - Involvement of hepatocyte nuclear factor 3 in endoderm differentiation of embryonic stem cells. AB - The transcription factors of the hepatocyte nuclear factor 3 (HNF3) family, which are active in the liver, are expressed early during endoderm differentiation. To study their involvement in early murine development, we examined their role in embryonic stem (ES) cells. HNF3alpha or HNF3beta mRNA transcripts were not detected in ES cells before differentiation, and only low levels of HNF3beta mRNA were detected at a late stage of differentiation of ES cells to embryoid bodies (EB) (20 days after induction of differentiation). To examine the consequences of overexpressing HNF3alpha or -beta in ES cells, we transfected the two genes into these cells and determined the levels of expression of tissue-specific genes during EB differentiation. Specifically, we examined expression of albumin, cystic fibrosis transmembrane conductance regulator (CFTR), phosphoenolpyruvate carboxykinase (PEPCK), alpha1-antitrypsin, transthyretin, zeta-globin, and neurofilament 68kd as markers for different cell lineages. Overexpression of HNF3beta (and to a lesser extent of HNF3alpha) induced the expression of genes associated with endodermal lineage, namely, the genes for CFTR and albumin, but did not induce the expression of genes involved in late endoderm differentiation, such as the genes for PEPCK and alpha1-antitrypsin. Moreover, expression of HNF1beta was highly induced in HNF3-overexpressing cells, while expression of HNF1alpha and HNF4 was only mildly induced in these cells. Therefore, HNF3alpha and -beta seem to be involved in early endoderm differentiation of ES cells and together with other developmental factors are apparently needed for the induction of the endodermal lineage in vivo. PMID- 9199316 TI - Expression and function of the leucine zipper protein Par-4 in apoptosis. AB - The prostate apoptosis response-4 (par-4) gene was identified by differential screening for genes that are upregulated when prostate cancer cells are induced to undergo apoptosis. The par-4 gene is induced by apoptotic signals but not by growth-arresting, necrotic, or growth-stimulatory signals. The deduced amino acid sequence of par-4 predicts a protein with a leucine zipper domain at its carboxy terminus. We have recently shown that the Par-4 protein binds, via its leucine zipper domain, to the zinc finger domain of Wilms' tumor protein WT1 (R. W. Johnstone et al., Mol. Cell. Biol. 16:6945-6956, 1996). In experiments aimed at determining the functional role of par-4 in apoptosis, an antisense par-4 oligomer abrogated par-4 expression and activator-driven apoptosis in rat prostate cancer cell line AT-3, suggesting that par-4 is required for apoptosis in these cells. Consistent with a functional role for par-4 in apoptosis, ectopic overexpression of par-4 in prostate cancer cell line PC-3 and melanoma cell line A375-C6 conferred supersensitivity to apoptotic stimuli. Transfection studies with deletion mutants of Par-4 revealed that full-length Par-4, but not mutants that lacked the leucine zipper domain of Par-4, conferred enhanced sensitivity to apoptotic stimuli. Most importantly, ectopic coexpression of the leucine zipper domain of Par-4 inhibited the ability of Par-4 to enhance apoptosis. Finally, ectopic expression of WT1 attenuated apoptosis, and coexpression of Par-4 but not a leucine zipperless mutant of Par-4 rescued the cells from the antiapoptotic effect of WT1. These findings suggest that the leucine zipper domain is required for the Par-4 protein to function in apoptosis. PMID- 9199317 TI - Beta interferon and oncostatin M activate Raf-1 and mitogen-activated protein kinase through a JAK1-dependent pathway. AB - Activation of early response genes by interferons (IFNs) and other cytokines requires tyrosine phosphorylation of a family of transcription factors termed signal transducers and activators of transcription (Stats). The Janus family of tyrosine kinases (Jak1, Jak2, Jak3, and Tyk2) is required for cytokine-induced tyrosine phosphorylation and dimerization of the Stat proteins. In order for IFNs to stimulate maximal expression of Stat1alpha-regulated genes, phosphorylation of a serine residue in the carboxy terminus by mitogen-activated protein kinase (MAPK) is also required. In HeLa cells, both IFN-beta and oncostatin M (OSM) stimulated MAPK and Raf-1 enzyme activity, in addition to Stat1 and Stat3 tyrosine phosphorylation. OSM stimulation of Raf-1 correlated with GTP loading of Ras, whereas IFN-beta activation of Raf-1 was Ras independent. IFN-beta- and OSM induced Raf-1 activity could be coimmunoprecipitated with either Jak1 or Tyk2. Furthermore, HeLa cells lacking Jak1 displayed no activation of STAT1alpha, STAT3, and Raf-1 by IFN-beta or OSM and also demonstrated no increase in the relative level of GTP-bound p21ras in response to OSM. The requirement for Jak1 for IFN-beta- and OSM-induced activation of Raf-1 was also seen in Jak1-deficient U4A fibrosarcoma cells. Interestingly, basal MAPK, but not Raf-1, activity was constitutively enhanced in Jak1-deficient HeLa cells. Transient expression of Jak1 in both Jak-deficient HeLa cells and U4A cells reconstituted the ability of IFN-beta and OSM to activate Raf-1 and decreased the basal activity of MAPK, while expression of a kinase-inactive form of the protein showed no effect. Moreover, U4A cells selected for stable expression of Jak1, or COS cells transiently expressing Jak1 or Tyk2 but not Jak3, exhibited enhanced Raf-1 activity. Therefore, it appears that Jak1 is required for Raf-1 activation by both IFN-beta and OSM. These results provide evidence for a link between the Jaks and the Raf/MAPK signaling pathways. PMID- 9199318 TI - Phosphatidylinositol 4,5-bisphosphate phosphatase regulates the rearrangement of actin filaments. AB - Phosphatidylinositol 4,5-bisphosphate (PIP2) reorganizes actin filaments by modulating the functions of a variety of actin-regulatory proteins. Until now, it was thought that bound PIP2 is hydrolyzed only by tyrosine-phosphorylated phospholipase Cgamma (PLCgamma) after the activation of tyrosine kinases. Here, we show a new mechanism for the hydrolysis of bound PIP2 and the regulation of actin filaments by PIP2 phosphatase (synaptojanin). We isolated a 150-kDa protein (p150) from brains that binds the SH3 domains of Ash/Grb2. The sequence of this protein was found to be homologous to that of synaptojanin. The expression of p150 in COS 7 cells produces a decrease in the number of actin stress fibers in the center of the cells and causes the cells to become multinuclear. On the other hand, the expression of a PIP2 phosphatase-negative mutant does not disrupt actin stress fibers or produce the multinuclear phenotype. We have also shown that p150 forms the complexes with Ash/Grb2 and epidermal growth factor (EGF) receptors only when the cells are treated with EGF and that it reorganizes actin filaments in an EGF-dependent manner. Moreover, the PIP2 phosphatase activity of native p150 purified from bovine brains is not inhibited by profilin, cofilin, or alpha actinin, although PLCdelta1 activity is markedly inhibited by these proteins. Furthermore, p150 suppresses actin gelation, which is induced by smooth muscle alpha-actinin. All these data suggest that p150 (synaptojanin) hydrolyzes PIP2 bound to actin regulatory proteins, resulting in the rearrangement of actin filaments downstream of tyrosine kinase and Ash/Grb2. PMID- 9199319 TI - Ras links growth factor signaling to the cell cycle machinery via regulation of cyclin D1 and the Cdk inhibitor p27KIP1. AB - Activation of growth factor receptors by ligand binding initiates a cascade of events leading to cell growth and division. Progression through the cell cycle is controlled by cyclin-dependent protein kinases (Cdks), but the mechanisms that link growth factor signaling to the cell cycle machinery have not been established. We report here that Ras proteins play a key role in integrating mitogenic signals with cell cycle progression through G1. Ras is required for cell cycle progression and activation of both Cdk2 and Cdk4 until approximately 2 h before the G1/S transition, corresponding to the restriction point. Analysis of Cdk-cyclin complexes indicates that Ras signaling is required both for induction of cyclin D1 and for downregulation of the Cdk inhibitor p27KIP1. Constitutive expression of cyclin D1 circumvents the requirement for Ras signaling in cell proliferation, indicating that regulation of cyclin D1 is a critical target of the Ras signaling cascade. PMID- 9199320 TI - DNA elements regulating alpha1-tubulin gene induction during regeneration of eukaryotic flagella. AB - Eukaryotic flagella are complex organelles composed of more than 200 polypeptides. Little is known about the regulatory mechanisms governing synthesis of the flagellar protein subunits and their assembly into this complex organelle. The unicellular green alga Chlamydomonas reinhardtii is the premier experimental model system for studying such cellular processes. When acid shocked, C. reinhardtii excises its flagella, rapidly and coordinately activates transcription of a set of flagellar genes, and ultimately regenerates a new flagellar pair. To define functionally the regulatory sequences that govern induction of the set of genes after acid shock, we analyzed the alpha1-tubulin gene promoter. To simplify transcriptional analysis in vivo, we inserted the selectable marker gene ARG7 on the same plasmid with a tagged alpha1-tubulin gene and stably introduced it into C. reinhardtii cells. By deletion of various sequences, two promoter regions (-176 to -122 and -85 to -16) were identified as important for induction of the tagged alpha1-tubulin gene. Deleting the region between -176 and -122 from the transcription start site resulted in an induction level which was only 45 to 70% of that of the resident gene. Deleting the region upstream of -56 resulted in a complete loss of inducibility without affecting basal expression. The alpha1-tubulin promoter region from -85 to -16 conferred partial acid shock inducibility to an arylsulfatase (ARS) reporter gene. These results show that induction of the alpha1-tubulin gene after acid shock is a complex response that requires diverse sequence elements. PMID- 9199321 TI - Specific regulation of E2F family members by cyclin-dependent kinases. AB - The transcription factor E2F-1 interacts stably with cyclin A via a small domain near its amino terminus and is negatively regulated by the cyclin A-dependent kinases. Thus, the activities of E2F, a family of transcription factors involved in cell proliferation, are regulated by at least two types of cell growth regulators: the retinoblastoma protein family and the cyclin-dependent kinase family. To investigate further the regulation of E2F by cyclin-dependent kinases, we have extended our studies to include additional cyclins and E2F family members. Using purified components in an in vitro system, we show that the E2F-1 DP-1 heterodimer, the functionally active form of the E2F activity, is not a substrate for the active cyclin D-dependent kinases but is efficiently phosphorylated by the cyclin B-dependent kinases, which do not form stable complexes with the E2F-1-DP-1 heterodimer. Phosphorylation of the E2F-1-DP-1 heterodimer by cyclin B-dependent kinases, however, did not result in down regulation of its DNA-binding activity, as is readily seen after phosphorylation by cyclin A-dependent kinases, suggesting that phosphorylation per se is not sufficient to regulate E2F DNA-binding activity. Furthermore, heterodimers containing E2F-4, a family member lacking the cyclin A binding domain found in E2F-1, are not efficiently phosphorylated or functionally down-regulated by cyclin A-dependent kinases. However, addition of the E2F-1 cyclin A binding domain to E2F-4 conferred cyclin A-dependent kinase-mediated down-regulation of the E2F-4-DP-1 heterodimer. Thus, both enzymatic phosphorylation and stable physical interaction are necessary for the specific regulation of E2F family members by cyclin-dependent kinases. PMID- 9199322 TI - Denaturation of the simian virus 40 origin of replication mediated by human replication protein A. AB - The initiation of simian virus 40 (SV40) replication requires recognition of the viral origin of replication (ori) by SV40 T antigen, followed by denaturation of ori in a reaction dependent upon human replication protein A (hRPA). To understand how origin denaturation is achieved, we constructed a 48-bp SV40 "pseudo-origin" with a central 8-nucleotide (nt) bubble flanked by viral sequences, mimicking a DNA structure found within the SV40 T antigen-ori complex. hRPA bound the pseudo-origin with similar stoichiometry and an approximately fivefold reduced affinity compared to the binding of a 48-nt single-stranded DNA molecule. The presence of hRPA not only distorted the duplex DNA flanking the bubble but also resulted in denaturation of the pseudo-origin substrate in an ATP independent reaction. Pseudo-origin denaturation occurred in 7 mM MgCl2, distinguishing this reaction from Mg2+-independent DNA-unwinding activities previously reported for hRPA. Tests of other single-stranded DNA-binding proteins (SSBs) revealed that pseudo-origin binding correlates with the known ability of these SSBs to support the T-antigen-dependent origin unwinding activity. Our results suggest that hRPA binding to the T antigen-ori complex induces the denaturation of ori including T-antigen recognition sequences, thus releasing T antigen from ori to unwind the viral DNA. The denaturation activity of hRPA has the potential to play a significant role in other aspects of DNA metabolism, including DNA repair. PMID- 9199323 TI - Requirements for localization of p130cas to focal adhesions. AB - p130cas (Cas) is an adapter protein that has an SH3 domain followed by multiple SH2 binding motifs in the substrate domain. It also contains a tyrosine residue and a proline-rich sequence near the C terminus, which are the binding sites for the SH2 and SH3 domains of Src kinase, respectively. Cas was originally identified as a major tyrosine-phosphorylated protein in v-Crk- and v-Src transformed cells. Subsequently, Cas was shown to be inducibly tyrosine phosphorylated upon integrin stimulation; it is therefore regarded as one of the focal adhesion proteins. Using an immunofluorescence study, we examined the subcellular localization of Cas and determined the regions required for its localization to focal adhesions. In nontransformed cells, Cas was localized predominantly to the cytoplasm and partially to focal adhesions. However, in 527F c-Src-transformed cells, Cas was localized mainly to podosomes, where the focal adhesion proteins are assembled. The localization of Cas to focal adhesions was also observed in cells expressing the kinase-negative 527F/295M-c-Src. A series of analyses with deletion mutants expressed in various cells revealed that the SH3 domain of Cas is necessary for its localization to focal adhesions in nontransformed cells while both the SH3 domain and the C-terminal Src binding domain of Cas are required in 527F-c-Src-transformed cells and fibronectin stimulated cells. In addition, the localization of Cas to focal adhesions was abolished in Src-negative cells. These results demonstrate that the SH3 domain of Cas and the association of Cas with Src kinase play a pivotal role in the localization of Cas to focal adhesions. PMID- 9199324 TI - Adhesion-dependent regulation of an A+U-rich element-binding activity associated with AUF1. AB - Monocyte adherence results in the rapid transcriptional activation and mRNA stabilization of numerous mediators of inflammation and tissue repair. While the enhancer and promoter elements associated with transcriptional activation have been studied, mechanisms linking adhesion, mRNA stabilization, and translation are unknown. GROalpha and interleukin-1beta (IL-1beta) mRNAs are highly labile in nonadhered monocytes but stabilize rapidly after adherence. GROalpha and IL-1beta transcripts both contain A+U-rich elements (AREs) in the 3' untranslated region (UTR) which have been directly associated with rapid mRNA turnover. To determine if the GROalpha ARE region was recognized by factors associated with mRNA degradation, we carried out mobility gel shift analyses using a series of RNA probes encompassing the entire GROalpha transcript. Stable complexes were formed only with the proximal 3' UTR which contained the ARE region. The two slower moving complexes were rapidly depleted following monocyte adherence but not direct integrin engagement. Deadherence reactivated the two largest ARE-binding complexes and destabilized IL-1beta transcripts. Antibody supershift studies demonstrated that both of these ARE RNA-binding complexes contained AUF1. The formation of these complexes and the accelerated mRNA turnover are phosphorylation-dependent events, as both are induced in adherent monocytes by the tyrosine kinase inhibitor genistein and the p38 MAP kinase inhibitor of IL 1beta translation, SK&F 86002. These results demonstrate that cell adhesion and deadhesion rapidly and reversibly modify both cytokine mRNA stability and the RNA binding complexes associated with AUF1. PMID- 9199325 TI - RNA recognition by the human polyadenylation factor CstF. AB - Polyadenylation of mammalian mRNA precursors requires at least two signal sequences in the RNA: the nearly invariant AAUAAA, situated 5' to the site of polyadenylation, and a much more variable GU- or U-rich downstream element. At least some downstream sequences are recognized by the heterotrimeric polyadenylation factor CstF, although how, and indeed if, all variations of this diffuse element are bound by a single factor is unknown. Here we show that the RNP-type RNA binding domain of the 64-kDa subunit of CstF (CstF-64) (64K RBD) is sufficient to define a functional downstream element. Selection-amplification (SELEX) experiments employing a glutathione S-transferase (GST)-64K RBD fusion protein selected GU-rich sequences that defined consensus recognition motifs closely matching those present in natural poly(A) sites. Selected sequences were bound specifically, and with surprisingly high affinities, by intact CstF and were functional in reconstituted, CstF-dependent cleavage assays. Our results also indicate that GU- and U-rich sequences are variants of a single CstF recognition motif. For comparison, SELEX was performed with a GST fusion containing the RBD from the apparent yeast homolog of CstF-64, RNA15. Strikingly, although the two RBDs are almost 50% identical and yeast poly(A) signals are at least as degenerate as the mammalian downstream element, a nearly invariant 12 base U-rich sequence distinct from the CstF-64 consensus was identified. We discuss these results in terms of the function and evolution of mRNA 3'-end signals. PMID- 9199326 TI - Mitochondrial glutamate dehydrogenase from Leishmania tarentolae is a guide RNA binding protein. AB - To identify specific proteins interacting with guide RNAs (gRNAs) in mitochondrial ribonucleoprotein complexes from Leishmania tarentolae, fractionated and unfractionated mitochondrial extracts were subjected to UV cross linking with added labeled gRNA and also with [alpha-32P]UTP-labeled endogenous RNA. An abundant 110-kDa protein (p110) localized in the T-V complex, which sediments in glycerol gradients at the leading edge of the 10S terminal uridylyltransferase peak, was found to interact with both types of labeled RNAs. The p110 protein was gel isolated and subjected to microsequence analysis, and the gene was cloned. The sequence revealed significant similarity with mitochondrial glutamate dehydrogenases. A polyclonal antiserum was raised against a recombinant fragment of the p110 gene and was used to demonstrate a stable and specific gRNA-binding activity by coimmunoprecipitation and competitive gel shift analyses. Complex formation was strongly inhibited by competition with poly(U) or by deletion or substitution of the gRNA 3' oligo(U) tail. Also, addition of a 3' oligo(U) tail to an unrelated transcript was sufficient for p110 binding. Both the gRNA-binding activity of the p110 protein and in vitro gRNA-independent and gRNA-dependent uridine insertion activities in the mitochondrial extract were inhibited by high concentrations of dinucleotides. PMID- 9199327 TI - Transcription enhancer factor 1 interacts with a basic helix-loop-helix zipper protein, Max, for positive regulation of cardiac alpha-myosin heavy-chain gene expression. AB - The M-CAT binding factor transcription enhancer factor 1 (TEF-1) has been implicated in the regulation of several cardiac and skeletal muscle genes. Previously, we identified an E-box-M-CAT hybrid (EM) motif that is responsible for the basal and cyclic AMP-inducible expression of the rat cardiac alpha-myosin heavy chain (alpha-MHC) gene in cardiac myocytes. In this study, we report that two factors, TEF-1 and a basic helix-loop-helix leucine zipper protein, Max, bind to the alpha-MHC EM motif. We also found that Max was a part of the cardiac troponin T M-CAT-TEF-1 complex even when the DNA template did not contain an apparent E-box binding site. In the protein-protein interaction assay, a stable association of Max with TEF-1 was observed when glutathione S-transferase (GST) TEF-1 or GST-Max was used to pull down in vitro-translated Max or TEF-1, respectively. In addition, Max was coimmunoprecipitated with TEF-1, thus documenting an in vivo TEF-1-Max interaction. In the transient transcription assay, overexpression of either Max or TEF-1 resulted a mild activation of the alpha-MHC-chloramphenicol acetyltransferase (CAT) reporter gene at lower concentrations and repression of this gene at higher concentrations. However, when Max and TEF-1 expression plasmids were transfected together, the repression mediated by a single expression plasmid was alleviated and a three- to fourfold transactivation of the alpha-MHC-CAT reporter gene was observed. This effect was abolished once the EM motif in the promoter-reporter construct was mutated, thus suggesting that the synergistic transactivation function of the TEF-1-Max heterotypic complex is mediated through binding of the complex to the EM motif. These results demonstrate a novel association between Max and TEF-1 and indicate a positive cooperation between these two factors in alpha-MHC gene regulation. PMID- 9199328 TI - Interdigitated residues within a small region of VP16 interact with Oct-1, HCF, and DNA. AB - Upon infection, the herpes simplex virus (HSV) activator of immediate-early (IE) gene transcription VP16 forms a multiprotein-DNA complex with two cellular proteins, Oct-1 and HCF. First, VP16 associates with HCF independently of DNA, and this association stimulates subsequent association with Oct-1 on the DNA target of VP16 activation, the TAATGARAT motif found in HSV IE promoters. We have analyzed the involvement of VP16 residues lying near the carboxy-terminal transcriptional activation domain of VP16 in associating with HCF, Oct-1, and DNA. To assay VP16 association with HCF, we developed an electrophoretic mobility retardation assay in which HCF is used to retard the mobility of a hybrid VP16 GAL4 DNA-binding domain fusion protein bound to a GAL4 DNA-binding site. Analysis of an extensive set of individual and combined alanine substitutions over a 61 amino-acid region of VP16 shows that, even within a region as small as 13 amino acids, there are separate residues involved in association with either HCF, DNA, or Oct-1 bound to DNA; indeed, of two immediately adjacent amino acids in VP16, one is important for DNA binding and the other is important for HCF binding. These results suggest that a small region in VP16 is important for linking in close juxtaposition the four components of the VP16-induced complex and support the hypothesis that the structure of the Oct-1-VP16 interaction in this complex is similar to that formed by the yeast transcriptional regulatory proteins MATa1 and MAT alpha2. We propose that HCF stabilizes this Oct-1-VP16 interaction. PMID- 9199329 TI - Mitogen-activated and cyclin-dependent protein kinases selectively and differentially modulate transcriptional enhancement by the glucocorticoid receptor. AB - Cyclin-dependent kinase (CDK) and mitogen-activated protein kinase (MAPK) phosphorylate the rat glucocorticoid receptor in vitro at distinct sites that together correspond to the major phosphorylated receptor residues observed in vivo; MAPK phosphorylates receptor residues threonine 171 and serine 246, whereas multiple CDK complexes modify serines 224 and 232. Mutations in these kinases have opposite effects on receptor transcriptional activity in vivo. Receptor dependent transcriptional enhancement is reduced in yeast strains deficient in the catalytic (p34CDC28) or certain regulatory (cyclin) subunits of CDK complexes and is increased in a strain devoid of the mammalian MAPK homologs FUS3 and KSS1. These findings indicate that the glucocorticoid receptor is a target for multiple kinases in vivo, which either positively or negatively regulate receptor transcriptional enhancement. The control of receptor transcriptional activity via phosphorylation provides an increased array of regulatory inputs that, in addition to steroid hormones, can influence receptor function. PMID- 9199330 TI - Transcription of the HS2 enhancer toward a cis-linked gene is independent of the orientation, position, and distance of the enhancer relative to the gene. AB - The locus control region (LCR) regulates transcription of the downstream beta like globin genes 10 to 50 kb away. Among hypersensitive sites HS4, -3, -2, and 1, which define the LCR in erythroid cells, HS2 possesses prominent enhancer function. The mechanism by which the HS2 enhancer and other functional components of the LCR act over the distance is not clear. We have used reverse transcription PCR and RNase protection assays to analyze the transcriptional statuses of both the endogenous and the transfected HS2 enhancer in erythroid K562 cells. A novel pattern of HS2 enhancer transcription was observed. The endogenous HS2 enhancer was transcribed predominantly in the direction toward the downstream globin genes. The HS2 enhancer in transfected recombinant chloramphenicol acetyltransferase (CAT) plasmids was also transcribed predominantly toward the CAT gene, regardless of whether the enhancer was placed (i) in the genomic or reverse genomic orientation, (ii) in a position 5' or 3' to the gene, or (iii) at various distances up to 6 kb from the gene. The orientation, position, and distance independence in gene-tropic transcription of the HS2 enhancer correlates with the observed orientation, position, and distance independence of HS2 enhancer function and suggests that enhancer transcription may play a role in enhancer function. PMID- 9199331 TI - Genetic analysis of regulatory mutants affecting synthesis of extracellular proteinases in the yeast Yarrowia lipolytica: identification of a RIM101/pacC homolog. AB - Depending on the pH of the growth medium, the yeast Yarrowia lipolytica secretes both an acidic proteinase and an alkaline proteinase, the synthesis of which is also controlled by carbon, nitrogen, and sulfur availability, as well as by the presence of extracellular proteins. Recessive mutations at four unlinked loci, named PAL1 to PAL4, were isolated which prevent alkaline proteinase derepression under conditions of carbon and nitrogen limitation at pH 6.8. These mutations markedly affect mating and sporulation. A dominant suppressor of all four PAL mutations was isolated from a wild-type genomic library, which turned out to be a C-terminally truncated form of a 585-residue transcriptional factor of the His2Cys2 zinc finger family, which we propose to call YlRim101p. Another C terminally truncated version of YlRim101p (419 residues) is encoded by the dominant RPH2 mutation previously isolated as expressing alkaline protease independently of the pH. YlRim101p is homologous to the transcriptional activators Rim101p of Saccharomyces cerevisiae, required for entry into meiosis, and PacC of Aspergillus nidulans and Penicillium chrysogenum, which were recently shown to mediate regulation by ambient pH. YlRim101p appears essential for mating and sporulation and for alkaline proteinase derepression. YlRIM101 expression is autoregulated, maximal at alkaline pH, and strongly impaired by PAL mutations. PMID- 9199332 TI - Heterodimeric interaction between retinoid X receptor alpha and orphan nuclear receptor OR1 reveals dimerization-induced activation as a novel mechanism of nuclear receptor activation. AB - OR1 is a member of the steroid/thyroid hormone nuclear receptor superfamily which has been described to mediate transcriptional responses to retinoids and oxysterols. On a DR4 response element, an OR1 heterodimer with the nuclear receptor retinoid X receptor alpha (RXR alpha) has been described to convey transcriptional activation in both the absence and presence of the RXR ligand 9 cis retinoic acid, the mechanisms of which have remained unclear. Here, we dissect the effects of RXR alpha and OR1 ligand-binding domain interaction on transcriptional regulation and the role of the respective carboxy-terminal activation domains (AF-2s) in the absence and presence of the RXR ligand, employing chimeras of the nuclear receptors containing the heterologous GAL4 DNA binding domain as well as natural receptors. The results show that the interaction of the RXR and OR1 ligand-binding domains unleashes a transcription activation potential that is mainly dependent on the AF-2 of OR1, indicating that interaction with RXR activates OR1. This defines dimerization-induced activation as a novel function of heterodimeric interaction and mechanism of receptor activation not previously described for nuclear receptors. Moreover, we present evidence that activation of OR1 occurs by a conformational change induced upon heterodimerization with RXR. PMID- 9199333 TI - Functional characterization of the transactivation properties of the PDX-1 homeodomain protein. AB - Pancreas formation is prevented in mice carrying a null mutation in the PDX-1 homeoprotein, demonstrating a key role for this factor in development. PDX-1 can also bind to and activate transcription from cis-acting regulatory sequences in the insulin and somatostatin genes, which are expressed in pancreatic islet beta and delta cells, respectively. In this study, we compared the functional properties of PDX-1 with those of the closely related Xenopus homeoprotein XIHbox8. Analysis of chimeras between PDX-1, XIHbox8, and the DNA-binding domain of the Saccharomyces cerevisiae transcription factor GAL4 revealed that their transactivation domain was contained within the N-terminal region (amino acids 1 to 79). Detailed mutagenesis of this region indicated that transactivation is mediated by three highly conserved sequences, spanning amino acids 13 to 22 (subdomain A), 32 to 38 (subdomain B), and 60 to 73 (subdomain C). These sequences were also required by PDX-1 to synergistically activate insulin enhancer-mediated transcription with another key insulin gene activator, the E2A encoded basic helix-loop-helix E2-5 and E47 proteins. These results indicated that N-terminal sequences conserved between the mammalian PDX-1 and Xenopus XIHbox8 proteins are important in transcriptional activation. Stable expression of the PDX-1 deltaABC mutant in the insulin- and PDX-1-expressing betaTC3 cell line resulted in a threefold reduction in the rate of endogenous insulin gene transcription. Strikingly, the level of the endogenous PDX-1 protein was reduced to very low levels in these cells. These results suggest that PDX-1 is not absolutely essential for insulin gene expression in betaTC3 cells. We discuss the possible significance of these findings for insulin gene transcription in islet beta cells. PMID- 9199334 TI - Nuclear receptor steroidogenic factor 1 directs embryonic stem cells toward the steroidogenic lineage. AB - The orphan nuclear receptor steroidogenic factor 1 (SF-1) is expressed in the adrenal gland and gonads and is an important regulator of the expression of cytochrome P-450 steroidogenic enzymes in cultured cells. Targeted disruption of the SF-1 gene in mice shows that it is a critical participant in the genetic program that promotes the development of urogenital mesoderm into the adrenal gland and gonads. To assess the ability of SF-1 to regulate this differentiation pathway, we ectopically expressed SF-1 in murine embryonic stem (ES) cells. We found that stable expression of SF-1 is sufficient to alter ES cell morphology, permit cyclic AMP (cAMP) and retinoic acid-induced expression of the endogenous side chain cleavage enzyme gene, and consequently, promote steroidogenesis. While steroid production is dependent upon SF-1, cAMP induction of steroidogenesis does not enhance the responsiveness of an SF-1-specific reporter. Furthermore, the activity of a P450SCC promoter/luciferase reporter construct, which is induced by cAMP in steroidogenic cells and ES cells converted by stable expression of SF-1, is not induced by cAMP in wild-type ES cells transiently transfected with SF-1, suggesting that the induction of downstream gene products is required before steroidogenesis can occur. We demonstrate that mutants which disrupt the DNA binding domain or the AF2 transcriptional activation domain of SF-1 do not confer the steroidogenic phenotype to ES cells. Interestingly, however, AF2 mutants fused to the VP16 activation domain do confer the steroidogenic phenotype to ES cells, but only in the presence of a portion of the ligand binding domain. These studies extend the role of SF-1 in steroidogenic tissues to that of a dominant regulator of the steroidogenic cell phenotype. PMID- 9199335 TI - Characterization of a neuregulin-related gene, Don-1, that is highly expressed in restricted regions of the cerebellum and hippocampus. AB - Members of the epidermal growth factor family of receptors have long been implicated in the pathogenesis of various tumors, and more recently, apparent roles in the developing heart and nervous system have been described. Numerous ligands that activate these receptors have been isolated. We report here on the cloning and initial characterization of a second ligand for the erbB family of receptors. This factor, which we have termed Don-1 (divergent of neuregulin 1), has structural similarity with the neuregulins. We have isolated four splice variants, two each from human and mouse, and have shown that they are capable of inducing tyrosine phosphorylation of erbB3, erbB4, and erbB2. In contrast to those of neuregulin, high levels of expression of Don-1 are restricted to the cerebellum and dentate gyrus in the adult brain and to fetal tissues. PMID- 9199336 TI - Involvement of interleukin-8, vascular endothelial growth factor, and basic fibroblast growth factor in tumor necrosis factor alpha-dependent angiogenesis. AB - Tumor necrosis factor alpha (TNF-alpha) is a macrophage/monocyte-derived polypeptide which modulates the expression of various genes in vascular endothelial cells and induces angiogenesis. However, the underlying mechanism by which TNF-alpha mediates angiogenesis is not completely understood. In this study, we assessed whether TNF-alpha-induced angiogenesis is mediated through TNF alpha itself or indirectly through other TNF-alpha-induced angiogenesis-promoting factors. Cellular mRNA levels of interleukin-8 (IL-8), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and their receptors were increased after the treatment of human microvascular endothelial cells with TNF-alpha (100 U/ml). TNF-alpha-dependent tubular morphogenesis in vascular endothelial cells was inhibited by the administration of anti-IL-8, anti-VEGF, and anti-bFGF antibodies, and coadministration of all three antibodies almost completely abrogated tubular formation. Moreover, treatment with Sp1, NF-kappaB, and c-Jun antisense oligonucleotides inhibited TNF-alpha-dependent tubular morphogenesis by microvascular endothelial cells. Administration of a NF-kappaB antisense oligonucleotide almost completely inhibited TNF-alpha-dependent IL-8 production and partially abrogated TNF-alpha-dependent VEGF production, and an Sp1 antisense sequence partially inhibited TNF-alpha-dependent production of VEGF. A c-Jun antisense oligonucleotide significantly inhibited TNF-alpha dependent bFGF production but did not affect the production of IL-8 and VEGF. Administration of an anti-IL-8 or anti-VEGF antibody also blocked TNF-alpha induced neovascularization in the rabbit cornea in vivo. Thus, angiogenesis by TNF-alpha appears to be modulated through various angiogenic factors, both in vitro and in vivo, and this pathway is controlled through paracrine and/or autocrine mechanisms. PMID- 9199337 TI - Analysis of the sorting signals directing NADH-cytochrome b5 reductase to two locations within yeast mitochondria. AB - Mitochondrial NADH-cytochrome b5 reductase (Mcr1p) is encoded by a single nuclear gene and imported into two different submitochondrial compartments: the outer membrane and the intermembrane space. We now show that the amino-terminal 47 amino acids suffice to target the Mcr1 protein to both destinations. The first 12 residues of this sequence function as a weak matrix-targeting signal; the remaining residues are mostly hydrophobic and serve as an intramitochondrial sorting signal for the outer membrane and the intermembrane space. A double point mutation within the hydrophobic region of the targeting sequence virtually abolishes the ability of the precursor to be inserted into the outer membrane but increases the efficiency of transport into the intermembrane space. Import of Mcr1p into the intermembrane space requires an electrochemical potential across the inner membrane, as well as ATP in the matrix, and is strongly impaired in mitochondria lacking Tom7p or Tim11p, two components of the translocation machineries in the outer and inner mitochondrial membranes, respectively. These results indicate that intramitochondrial sorting of the Mcr1 protein is mediated by specific interactions between the bipartite targeting sequence and components of both mitochondrial translocation systems. PMID- 9199338 TI - Small heat shock protein suppression of Vpr-induced cytoskeletal defects in budding yeast. AB - Expression of the auxiliary human immunodeficiency virus type 1 (HIV-1) protein Vpr causes arrest of primate host cells in G2. Expression of this protein in budding yeast has been previously reported to cause growth arrest and a large cell phenotype. Investigation of the effect of Vpr expression in budding yeast, reported here, showed that it causes disruption of the actin cytoskeleton. Expression of HSP42, the gene for a small heat shock protein (sHSP), from a high copy-number plasmid reversed this effect. The sHSPs are induced by exposure of cells to thermal, osmotic, and oxidative stresses and to mitogens. In animal cells, overexpression of sHSPs causes increased resistance to stress and stabilization of actin stress fibers. Yeast cells subjected to mild stress, such as shifting from 23 to 39 degrees C, arrest growth and then resume cell division. Growth arrest is accompanied by transient disorganization of the cytoskeleton. Yeast in which the HSP42 gene was disrupted and which was subjected to moderate thermal stress reorganized the actin cytoskeleton more slowly than did wild-type control cells. These results demonstrate that in yeast, as in metazoan cells, sHSPs promote maintenance of the actin cytoskeleton. PMID- 9199339 TI - Precise switching of DNA replication timing in the GC content transition area in the human major histocompatibility complex. AB - The human genome is composed of long-range G+C% (GC%) mosaic structures thought to be related to chromosome bands. We previously reported a boundary of megabase sized GC% mosaic domains at the junction area between major histocompatibility complex (MHC) classes II and III, proposing it as a possible chromosome band boundary. DNA replication timing during the S phase is known to be correlated cytogenetically with chromosome band zones, and thus the band boundaries have been predicted to contain a switch point for DNA replication timing. In this study, to identify to the nucleotide sequence level the replication switch point during the S phase, we determined the precise DNA replication timing for MHC classes II and III, focusing on the junction area. To do this, we used PCR-based quantitation of nascent DNA obtained from synchronized human myeloid leukemia HL60 cells. The replication timing changed precisely in the boundary region with a 2-h difference between the two sides, supporting the prediction that this region may be a chromosome band boundary. We supposed that replication fork movement terminates (pauses) or significantly slows in the switch region, which contains dense Alu clusters; polypurine/polypyrimidine tracts; di-, tri-, or tetranucleotide repeats; and medium-reiteration-frequency sequences. Because the nascent DNA in the switch region was recovered at low efficiency, we investigated whether this region is associated with the nuclear scaffold and found three scaffold-associated regions in and around the switch region. PMID- 9199340 TI - CD28 mediates transcriptional upregulation of the interleukin-2 (IL-2) promoter through a composite element containing the CD28RE and NF-IL-2B AP-1 sites. AB - Mutagenesis studies have demonstrated the requirement for the CD28-responsive element (CD28RE) within the interleukin-2 (IL-2) promoter for transcriptional upregulation by CD28. Here, we demonstrate that CD28 responsiveness is conferred by a composite element containing both the CD28RE and the NF-IL-2B AP-1 sites (RE/AP). Mutations at either site within the RE/AP composite element abolish activity. The RE/AP composite element is a site for signal integration within the IL-2 promoter, since its activation is dependent on at least two separate signalling pathways being activated, through the T-cell receptor, CD28, and/or phorbol myristate acetate. Activation is maximal when all three signals occur simultaneously. By using a panel of CD28 cytoplasmic domain mutants, it was found that the transcriptional activation of the RE/AP composite element correlates exactly with the pattern of IL-2 secretion induced by these mutants upon stimulation. Similar to the upregulation of IL-2 secretion, the transcriptional upregulation of the RE/AP composite element by CD28 is FK506 insensitive. The pattern of activation of the RE/AP composite element is different from that observed for either an NFAT or consensus AP-1 site, implying that RE/AP represents a unique element. Using gel shift analysis, we demonstrate that stimulation by CD28 induces the association of the NF-kappaB family member c-Rel to the CD28RE within the RE/AP composite element. The transcriptional upregulation of IL-2 by CD28 appears, therefore, to be mediated through the RE/AP composite element, involving the association of c-Rel with the CD28RE. PMID- 9199341 TI - Estrogen-dependent cyclin E-cdk2 activation through p21 redistribution. AB - In order to elucidate the mechanisms by which estrogens and antiestrogens modulate the growth of breast cancer cells, we have characterized the changes induced by estradiol that occur during the G1 phase of the cell cycle of MCF-7 human mammary carcinoma cells. Addition of estradiol relieves the cell cycle block created by tamoxifen treatment, leading to marked activation of cyclin E cdk2 complexes and phosphorylation of the retinoblastoma protein within 6 h. Cyclin D1 levels increase significantly while the levels of cyclin E, cdk2, and the p21 and p27 cdk inhibitors are relatively constant. However, the p21 cdk inhibitor shifts from its association with cyclin E-cdk2 to cyclin D1-cdk4, providing an explanation for the observed activation of the cyclin E-cdk2 complexes. These results support the notion that cyclin D1 has an important role in steroid-dependent cell proliferation and that estrogen, by regulating the activities of G1 cyclin-dependent kinases, can control the proliferation of breast cancer cells. PMID- 9199342 TI - DNA cleavage within the MLL breakpoint cluster region is a specific event which occurs as part of higher-order chromatin fragmentation during the initial stages of apoptosis. AB - A distinct population of therapy-related acute myeloid leukemia (t-AML) is strongly associated with prior administration of topoisomerase II (topo II) inhibitors. These t-AMLs display distinct cytogenetic alterations, most often disrupting the MLL gene on chromosome 11q23 within a breakpoint cluster region (bcr) of 8.3 kb. We recently identified a unique site within the MLL bcr that is highly susceptible to DNA double-strand cleavage by classic topo II inhibitors (e.g., etoposide and doxorubicin). Here, we report that site-specific cleavage within the MLL bcr can be induced by either catalytic topo II inhibitors, genotoxic chemotherapeutic agents which do not target topo II, or nongenotoxic stimuli of apoptotic cell death, suggesting that this site-specific cleavage is part of a generalized cellular response to an apoptotic stimulus. We also show that site-specific cleavage within the MLL bcr can be linked to the higher-order chromatin fragmentation that occurs during the initial stages of apoptosis, possibly through cleavage of DNA loops at their anchorage sites to the nuclear matrix. In addition, we show that site-specific cleavage is conserved between species, as specific DNA cleavage can also be demonstrated within the murine MLL locus. Lastly, site-specific cleavage during apoptosis can also be identified at the AML1 locus, a locus which is also frequently involved in chromosomal rearrangements present in t-AML patients. In conclusion, these results suggest the potential involvement of higher-order chromatin fragmentation which occurs as a part of a generalized apoptotic response in a mechanism leading to chromosomal translocation of the MLL and AML1 genes and subsequent t-AML. PMID- 9199343 TI - Proviral inactivation of the Npat gene of Mpv 20 mice results in early embryonic arrest. AB - The Mpv 20 transgenic mouse strain was created by infection of embryos with a defective retrovirus. When Mpv 20 heterozygous animals were crossed, no homozygous neonatal mice or midgestation embryos were identified. When embryos from heterozygous crosses were cultured in vitro, approximately one quarter arrested as uncompacted eight-cell embryos, indicating that proviral insertion resulted in a recessive lethal defect whose phenotype was manifest very early in development. Molecular cloning of the Mpv 20 insertion site revealed that the provirus had disrupted the Npat gene, a gene of unknown function, resulting in the production of a truncated Npat mRNA. Expression of the closely linked Atm gene was found to be unaffected by the provirus. PMID- 9199344 TI - Shc contains two Grb2 binding sites needed for efficient formation of complexes with SOS in B lymphocytes. AB - Cross-linking of the B-cell antigen receptor (BCR) induces tyrosine phosphorylation of Shc, which is believed to lead to the activation of Ras. Previous work has shown that tyrosine-phosphorylated Shc forms complexes with another adapter protein, Grb2, and the Ras guanine nucleotide exchange factor SOS. Here, we demonstrate that phosphorylation of Shc by the hematopoietic cell specific tyrosine kinase Syk induces binding of Grb2 to Shc, suggesting that Syk phosphorylates Shc in stimulated B cells. Surprisingly, Syk-phosphorylated Shc possesses two Grb2 binding sites rather than the one site that has been previously reported. Both of these sites are required for efficient formation of Shc-Grb2-SOS complexes in vitro and in vivo. We suggest that two Grb2 proteins anchored by a single Shc protein bind simultaneously to one SOS molecule, resulting in a complex that is more stable than a complex containing only a single Grb2 protein bound to one SOS molecule. This model is consistent with our observation that BCR stimulation greatly increases the amount of SOS associated with Grb2. PMID- 9199345 TI - HRS/SRp40-mediated inclusion of the fibronectin EIIIB exon, a possible cause of increased EIIIB expression in proliferating liver. AB - Serine-arginine (SR)-rich proteins are believed to be important in mediating alternative pre-mRNA splicing. HRS/SRp40 expression is elevated in liver cell proliferation during development, regeneration, and oncogenesis. We tested whether HRS expression correlates with the appearance of alternatively spliced fibronectin transcripts during liver growth. HRS was highly expressed during the proliferative phase of liver development, correlating with expression of the fibronectin EIIIB alternative exon. In regenerating liver, HRS protein was induced in a time course consistent with the observed increase in fibronectin transcripts containing the EIIIB exon, particularly in nonparenchymal liver cells. Furthermore, in an in vivo assay, HRS, and not other SR proteins, directly mediated EIIIB exon inclusion in the fibronectin transcript. This alternative splicing was dependent on a purine-rich region within the EIIIB exon to which HRS specifically bound. We have established that HRS has the potential to contribute to the regulation of fibronectin pre-mRNA splicing during liver growth. Changes in fibronectin forms may be important in modifying liver architecture during the proliferative response, thus providing a potential mechanism by which SR proteins may participate in cellular growth control. PMID- 9199346 TI - RING1 is associated with the polycomb group protein complex and acts as a transcriptional repressor. AB - The Polycomb (Pc) protein is a component of a multimeric, chromatin-associated Polycomb group (PcG) protein complex, which is involved in stable repression of gene activity. The identities of components of the PcG protein complex are largely unknown. In a two-hybrid screen with a vertebrate Pc homolog as a target, we identify the human RING1 protein as interacting with Pc. RING1 is a protein that contains the RING finger motif, a specific zinc-binding domain, which is found in many regulatory proteins. So far, the function of the RING1 protein has remained enigmatic. Here, we show that RING1 coimmunoprecipitates with a human Pc homolog, the vertebrate PcG protein BMI1, and HPH1, a human homolog of the PcG protein Polyhomeotic (Ph). Also, RING1 colocalizes with these vertebrate PcG proteins in nuclear domains of SW480 human colorectal adenocarcinoma and Saos-2 human osteosarcoma cells. Finally, we show that RING1, like Pc, is able to repress gene activity when targeted to a reporter gene. Our findings indicate that RING1 is associated with the human PcG protein complex and that RING1, like PcG proteins, can act as a transcriptional repressor. PMID- 9199347 TI - Localization and posttranslational modifications of otefin, a protein required for vesicle attachment to chromatin, during Drosophila melanogaster development. AB - Otefin is a peripheral protein of the inner nuclear membrane in Drosophila melanogaster. Here we show that during nuclear assembly in vitro, it is required for the attachment of membrane vesicles to chromatin. With the exception of sperm cells, otefin colocalizes with lamin Dm0 derivatives in situ and presumably in vivo and is present in all somatic cells examined during the different stages of Drosophila development. In the egg chamber, otefin accumulates in the cytoplasm, in the nuclear periphery, and within the nucleoplasm of the oocyte, in a pattern similar to that of lamin Dm0 derivatives. There is a relatively large nonnuclear pool of otefin present from stages 6 to 7 of egg chamber maturation through 6 to 8 h of embryonic development at 25 degrees C. In this pool, otefin is peripherally associated with a fraction containing the membrane vesicles. This association is biochemically different from the association of otefin with the nuclear envelope. Otefin is a phosphoprotein in vivo and is a substrate for in vitro phosphorylation by cdc2 kinase and cyclic AMP-dependent protein kinase. A major site for cdc2 kinase phosphorylation in vitro was mapped to serine 36 of otefin. Together, our data suggest an essential role for otefin in the assembly of the Drosophila nuclear envelope. PMID- 9199348 TI - The rRNA-processing function of the yeast U14 small nucleolar RNA can be rescued by a conserved RNA helicase-like protein. AB - The phylogenetically conserved U14 small nucleolar RNA is required for processing of rRNA, and this function involves base pairing with conserved complementary sequences in 18S RNA. With a view to identifying other important U14 interactions, a stem-loop domain required for activity of Saccharomyces cerevisiae U14 RNAs (the Y domain) was first subjected to detailed mutational analysis. The mapping results showed that most nucleotides of the Y domain can be replaced without affecting function, except for loop nucleotides conserved among five different yeast species. Defective variants were then used to identify both intragenic and extragenic suppressor mutations. All of the intragenic mutations mapped within six nucleotides of the primary mutation, suggesting that suppression involves a change in conformation and that the loop element is involved in an essential intermolecular interaction rather than intramolecular base pairing. A high-copy extragenic suppressor gene, designated DBP4 (DEAD box protein 4), encodes an essential, putative RNA helicase of the DEAD-DEXH box family. Suppression by DBP4 (initially CA4 [T.-H. Chang, J. Arenas, and J. Abelson, Proc. Natl. Acad. Sci. USA 87:1571-1575, 1990]) restores the level of 18S rRNA and is specific for the Y domain but is not allele specific. DBP4 is predicted to function either in assembly of the U14 small nucleolar RNP or, more likely, in its interaction with other components of the rRNA processing apparatus. Mediating the interaction of U14 with precursor 18S RNA is an especially attractive possibility. PMID- 9199349 TI - A novel transcript encoding an N-terminally truncated AML1/PEBP2 alphaB protein interferes with transactivation and blocks granulocytic differentiation of 32Dcl3 myeloid cells. AB - The gene AML1/PEBP2 alphaB encodes the alpha subunit of transcription factor PEBP2/CBF and is essential for the establishment of fetal liver hematopoiesis. Rearrangements of AML1 are frequently associated with several types of human leukemia. Three types of AML1 cDNA isoforms have been described to date; they have been designated AML1a, AML1b, and AML1c. All of these isoforms encode the conserved-Runt domain, which harbors the DNA binding and heterodimerization activities. We have identified a new isoform of the AML1 transcript, termed AML1 deltaN, in which exon 1 is directly connected to exon 4 by alternative splicing. The AML1 deltaN transcript was detected in various hematopoietic cell lines of lymphoid to myeloid cell origin, as revealed by RNase protection and reverse transcriptase PCR analyses. The protein product of AML1 deltaN lacks the N terminal region of AML1, including half of the Runt domain, and neither binds to DNA nor heterodimerizes with the beta subunit. However, AML1 deltaN was found to interfere with the transactivation activity of PEBP2, and the molecular region responsible for this activity was identified. Stable expression of AML1 deltaN in 32Dcl3 myeloid cells blocked granulocytic differentiation in response to granulocyte colony-stimulating factor. These results suggest that AML1 deltaN acts as a modulator of AML1 function and serves as a useful tool to dissect the functional domains in the C-terminal region of AML1. PMID- 9199350 TI - Regulation of the protein kinase PKR by the vaccinia virus pseudosubstrate inhibitor K3L is dependent on residues conserved between the K3L protein and the PKR substrate eIF2alpha. AB - The mammalian double-stranded RNA-activated protein kinase PKR is a component of the cellular antiviral defense mechanism and phosphorylates Ser-51 on the alpha subunit of the translation factor eIF2 to inhibit protein synthesis. To identify the molecular determinants that specify substrate recognition by PKR, we performed a mutational analysis on the vaccinia virus K3L protein, a pseudosubstrate inhibitor of PKR. High-level expression of PKR is lethal in the yeast Saccharomyces cerevisiae because PKR phosphorylates eIF2alpha and inhibits protein synthesis. We show that coexpression of vaccinia virus K3L can suppress the growth-inhibitory effects of PKR in yeast, and using this system, we identified both loss-of-function and hyperactivating mutations in K3L. Truncation of, or point mutations within, the C-terminal portion of the K3L protein, homologous to residues 79 to 83 in eIF2alpha, abolished PKR inhibitory activity, whereas the hyperactivating mutation, K3L-H47R, increased the homology between the K3L protein and eIF2alpha adjacent to the phosphorylation site at Ser-51. Biochemical and yeast two-hybrid analyses revealed that the suppressor phenotype of the K3L mutations correlated with the affinity of the K3L protein for PKR and was inversely related to the level of eIF2alpha phosphorylation in the cell. These results support the idea that residues conserved between the pseudosubstrate K3L protein and the authentic substrate eIF2alpha play an important role in substrate recognition, and they suggest that PKR utilizes sequences both near and over 30 residues from the site of phosphorylation for substrate recognition. Finally, by reconstituting part of the mammalian antiviral defense mechanism in yeast, we have established a genetically useful system to study viral regulators of PKR. PMID- 9199351 TI - Retinoid antagonism of NF-IL6: insight into the mechanism of antiproliferative effects of retinoids in Kaposi's sarcoma. AB - All-trans-retinoic acid (RA) is active in the treatment of Kaposi's sarcoma (KS), and retinoids inhibit KS cell growth in vitro. To understand the mechanism of retinoid action in KS, we studied the expression of autocrine growth factors of KS cells after RA treatment. We demonstrate that RA and its synthetic analogs inhibit the proliferation of KS cells by inhibiting the mRNA and protein levels of interleukin-6 (IL-6), an autocrine growth factor for KS cells. We further demonstrate that nuclear retinoid receptors (RA receptors [RARs] and retinoid X receptors [RXRs]) inhibit IL-6 promoter action by antagonizing the enhancer action of NF-IL6, a basic domain leucine zipper transcription factor belonging to the family of CAAT enhancer binding proteins. Furthermore, RARs and RXRs do not bind in vitro to an NF-IL6 binding site. However, the secondary folded structure of the DNA binding domain of RAR and RXR is obligatory for inhibiting NF-IL6 activity. Thus, NF-IL6 is a potential therapeutic target for the treatment of KS. Finally, using receptor-selective synthetic retinoids, we demonstrate that NF-IL6 antagonism and transactivation are separable functions of RAR alpha, thus indicating that synthetic retinoids with properties of NF-IL6 antagonism but lacking transactivation capabilities can be synthesized. Such retinoids might increase therapeutic potential in KS. PMID- 9199352 TI - Chimeras of the native form or achondroplasia mutant (G375C) of human fibroblast growth factor receptor 3 induce ligand-dependent differentiation of PC12 cells. AB - Mutations in the gene for human fibroblast growth factor receptor 3 (hFGFR3) cause a variety of skeletal dysplasias, including the most common genetic form of dwarfism, achondroplasia (ACH). Evidence indicates that these phenotypes are not due to simple haploinsufficiency of FGFR3 but are more likely related to a role in negatively regulating skeletal growth. The effects of one of these mutations on FGFR3 signaling were examined by constructing chimeric receptors composed of the extracellular domain of human platelet-derived growth factor receptor beta (hPDGFR beta) and the transmembrane and intracellular domains of hFGFR3 or of an ACH (G375C) mutant. Following stable transfection in PC12 cells, which lack platelet-derived growth factor (PDGF) receptors, all clonal cell lines, with either type of chimera, showed strong neurite outgrowth in the presence of PDGF but not in its absence. Antiphosphotyrosine immunoblots showed ligand-dependent autophosphorylation, and both receptor types stimulated strong phosphorylation of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase, an event associated with the differentiative response of these cells. In addition, ligand-dependent phosphorylation of phospholipase Cgamma and Shc was also observed. All of these responses were comparable to those observed from ligand activation, such as by nerve growth factor, of the native PC12 cells used to prepare the stable transfectants. The cells with the chimera bearing the ACH mutation were more rapidly responsive to ligand with less sustained MAPK activation, indicative of a preactivated or primed condition and consistent with the view that these mutations weaken ligand control of FGFR3 function. However, the full effect of the mutation likely depends in part on structural features of the extracellular domain. Although FGFR3 has been suggested to act as a negative regulator of long-bone growth in chrondrocytes, it produces differentiative signals similar to those of FGFR1, to which only positive effects have been ascribed, in PC12 cells. Therefore, its regulatory effects on bone growth likely result from cellular contexts and not the induction of a unique FGFR3 signaling pathway. PMID- 9199354 TI - The composition of coding joints formed in V(D)J recombination is strongly affected by the nucleotide sequence of the coding ends and their relationship to the recombination signal sequences. AB - V(D)J recombination proceeds in two stages. Precise cleavage at the border of the conserved recombination signal sequences (RSSs) and the coding ends results in flush double-stranded signal ends and coding ends terminating in hairpins. In the second stage, the signal and coding ends are processed into signal and coding joints. Coding ends containing certain nucleotide homopolymers affect the efficiency of V(D)J recombination. In this study, we have tested the effect of small changes in coding-end nucleotide composition on the frequency of coding- and signal joint formation. Furthermore, we have determined the sequences of coding joints resulting from recombination of coding ends with different compositions. We found that the presence of two T nucleotides 5' of both RSSs, but not a single T, reduces the frequency of signal joint formation, i.e., interferes with the cleavage stage of V(D)J recombination. However, coding-joint processing is sensitive even to a single T. Both the sequence of the coding ends and the particular RSS (12-mer or 23-mer) with which the coding end is associated affect the final composition of the coding joints. Thus, the presence of P nucleotides, the conservation of one undeleted coding end, the formation of joints without any deletions, and the template-dependent insertion of nucleotides are strongly influenced by the coding-end nucleotide composition and/or RSS association. The implications of these results with respect to the processing of coding ends are discussed. PMID- 9199355 TI - Establishment of the axis in chordates: facts and speculations. AB - A master plan for the early development of all chordates is proposed. The radial symmetry of the chordate ovum is changed at or after fertilization into a bilateral symmetry by an external signal. Until now two alternative triggers, sperm entry and gravity, have been demonstrated. It is suggested that a correlation exists between the amount of yolk stored in the egg and the mechanism used for axialization. The speed at which axialization of the embryo proper takes place depends on the translocation speed of maternal determinants from the vegetal pole towards the future dorsoposterior side of the embryo. On arrival at their destination, the activated determinants form, in all chordates, an induction center homologous to the amphibian 'Nieuwkoop center', which induces the formation of 'Spemann's organizer'. On the basis of the above general scenario, a revision is proposed of the staging of some embryonic types, as well as of the identification of germ layer and the spaces between them. PMID- 9199353 TI - The Saccharomyces cerevisiae DNA polymerase alpha catalytic subunit interacts with Cdc68/Spt16 and with Pob3, a protein similar to an HMG1-like protein. AB - We have used DNA polymerase alpha affinity chromatography to identify factors involved in eukaryotic DNA replication in the yeast Saccharomyces cerevisiae. Two proteins that bound to the catalytic subunit of DNA polymerase alpha (Pol1 protein) are encoded by the essential genes CDC68/SPT16 and POB3. The binding of both proteins was enhanced when extracts lacking a previously characterized polymerase binding protein, Ctf4, were used. This finding suggests that Cdc68 and Pob3 may compete with Ctf4 for binding to Pol1. Pol1 and Pob3 were coimmunoprecipitated from whole-cell extracts with antiserum directed against Cdc68, and Pol1 was immunoprecipitated from whole-cell extracts with antiserum directed against the amino terminus of Pob3, suggesting that these proteins may form a complex in vivo. CDC68 also interacted genetically with POL1 and CTF4 mutations; the maximum permissive temperature of double mutants was lower than for any single mutant. Overexpression of Cdc68 in a pol1 mutant strain dramatically decreased cell viability, consistent with the formation or modulation of an essential complex by these proteins in vivo. A mutation in CDC68/SPT16 had previously been shown to cause pleiotropic effects on the regulation of transcription (J. A. Prendergrast et al., Genetics 124:81-90, 1990; E. A. Malone et al., Mol. Cell. Biol. 11:5710-5717, 1991; A. Rowley et al., Mol. Cell. Biol. 11:5718-5726, 1991), with a spectrum of phenotypes similar to those caused by mutations in the genes encoding histone proteins H2A and H2B (Malone et al., Mol. Cell. Biol. 11:5710-5717, 1991). We show that at the nonpermissive temperature, cdc68-1 mutants arrest as unbudded cells with a 1C DNA content, consistent with a possible role for Cdc68 in the prereplicative stage of the cell cycle. The cdc68-1 mutation caused elevated rates of chromosome fragment loss, a phenotype characteristic of genes whose native products are required for normal DNA metabolism. However, this mutation did not affect the rate of loss or recombination for two intact chromosomes, nor did it affect the retention of a low-copy-number plasmid. The previously uncharacterized Pob3 sequence has significant amino acid sequence similarity with an HMG1-like protein from vertebrates. Based on these results and because Cdc68 has been implicated as a regulator of chromatin structure, we postulate that polymerase alpha may interact with these proteins to gain access to its template or to origins of replication in vivo. PMID- 9199356 TI - Cell movements, neuronal organisation and gene expression in hindbrains lacking morphological boundaries. AB - Rhombomeres are segmental units of the hindbrain that are separated from each other by a specialised zone of boundary cells. Retinoic acid application to a recently segmented hindbrain leads to disappearance of posterior rhombomere boundaries. Boundary loss is preceded by changes in segmental expression of Krox 20 and Cek-8 and followed by alterations in Hox gene expression. The characteristic morphology of boundary cells, their expression of follistatin and the periodic accumulation of axons normally associated with boundaries are all lost. In the absence of boundaries, we detect no change in anteroposterior dispersal of precursor cells and, in most cases, no substantial cell mixing between former rhombomeric units. This is consistent with the idea that lineage restriction can be maintained by processes other than a mechanical barrier composed of boundary cells. Much of the early organisation of the motor nuclei appears normal despite the loss of boundaries and altered Hox expression. PMID- 9199357 TI - Glial development in the Drosophila CNS requires concomitant activation of glial and repression of neuronal differentiation genes. AB - Two classes of glial cells are found in the embryonic Drosophila CNS, midline glial cells and lateral glial cells. Midline glial development is triggered by EGF-receptor signalling, whereas lateral glial development is controlled by the gcm gene. Subsequent glial cell differentiation depends partly on the pointed gene. Here we describe a novel component required for all CNS glia development. The tramtrack gene encodes two zinc-finger proteins, one of which, ttkp69, is expressed in all non-neuronal CNS cells. We show that ttkp69 is downstream of gcm and can repress neuronal differentiation. Double mutant analysis and coexpression experiments indicate that glial cell differentiation may depend on a dual process, requiring the activation of glial differentiation by pointed and the concomitant repression of neuronal development by tramtrack. PMID- 9199358 TI - The role of Engrailed in establishing the dorsoventral axis of the chick limb. AB - Expression and mutation analyses in mice suggest that the homeobox-containing gene Engrailed (En) plays a role in dorsoventral patterning of the limb. During the initial stages of limb bud outgrowth, En-1 mRNA and protein are uniformly distributed throughout the ventral limb bud ectoderm. Limbs of En-1(-/-) mice display a double dorsal phenotype suggesting that normal expression of En-1 in the ventral ectoderm is required to establish and/or maintain ventral limb characteristics. Loss of En-1 function also results in ventral expansion of the apical ectodermal ridge (AER), suggesting that En-1 is also required for proper formation of the AER. To further investigate the role En plays in dorsoventral patterning and AER formation, we have used the replication competent retroviral vector, RCAS, to mis-express mouse En-1 in the early chick limb bud. We show that ectopic En-1 expression in dorsal ectoderm is sufficient to repress the endogenous expression of the dorsal ectodermal marker Wnt7a, with a resultant decrease in Lmx1 expression in underlying dorsal mesenchyme. Furthermore, the AER is disrupted morphologically and the expression patterns of the AER signalling molecules Fgf-8 and Fgf-4 are altered. Consistent with recent evidence that there is a reciprocal interaction between signalling molecules in the dorsal ectoderm, AER, and zone of polarising activity (ZPA), loss of Wnt7a, Fgf-8 and Fgf-4 expression leads to a decrease in expression of the signalling molecule Shh in the ZPA. These results strongly support the idea that, in its normal domain of expression, En-1 represses Wnt7a-mediated dorsal differentiation by limiting the expression of Wnt7a to the dorsal ectoderm. Furthermore, our results provide additional evidence that En-1 is involved in AER formation and suggest that En-1 may act to define ventral ectodermal identity. PMID- 9199359 TI - Bmp-4 acts as a morphogen in dorsoventral mesoderm patterning in Xenopus. AB - The marginal zone is a ring of tissue that gives rise to a characteristic dorsoventral pattern of mesoderm in amphibian embryos. Bmp-4 is thought to play an important role in specifying ventral mesodermal fate. Here we show (1) that different doses of Bmp-4 are sufficient to pattern four distinct mesodermal cell types and to pattern gene expression in the early gastrula marginal zone into three domains, (2) that there is a graded requirement for a Bmp signal in mesodermal patterning, and (3) that Bmp-4 has long-range activity which can become graded in the marginal zone by the antagonizing action of noggin. The results argue that Bmp-4 acts as a morphogen in dorsoventral patterning of mesoderm. PMID- 9199360 TI - Dorsoventral patterning of the vertebrate neural tube is conserved in a protochordate. AB - The notochord and dorsal ectoderm induce dorsoventral compartmentalization of the vertebrate neural tube through the differential regulation of genes such as HNF 3beta, Pax3, Pax6 and snail. Here we analyze the expression of HNF-3beta and snail homologues in the ascidian, Ciona intestinalis, a member of the subphylum Urochordata, the earliest branch in the chordate phylum. A combination of in situ hybridization and promoter fusion analyses was used to demonstrate that the Ciona HNF-3beta homologue is expressed in the ventralmost ependymal cells of the neural tube, while the Ciona snail homologue is expressed at the junction between the invaginating neuroepithelium and dorsal ectoderm, similar to the patterns seen in vertebrates. These findings provide evidence that dorsoventral compartmentalization of the chordate neural tube is not an innovation of the vertebrates. We propose that precursors of the floor plate and neural crest were present in a common ancestor of both vertebrates and ascidians. PMID- 9199361 TI - Muller-cell-derived leukaemia inhibitory factor arrests rod photoreceptor differentiation at a postmitotic pre-rod stage of development. AB - In the present study, we examine rod photoreceptor development in dissociated cell cultures of neonatal mouse retina. We show that, although very few rhodopsin+ rods develop in the presence of 10% foetal calf serum (FCS), large numbers develop in the absence of serum, but only if the cell density in the cultures is high. The rods all develop from nondividing rhodopsin- cells, and new rods continue to develop from rhodopsin- cells for at least 6-8 days, indicating that there can be a long delay between when a precursor cell withdraws from the cell cycle and when it becomes a rhodopsin+ rod. We show that FCS arrests rod development in these cultures at a postmitotic, rhodopsin-, pre-rod stage. We present evidence that FCS acts indirectly by stimulating the proliferation of Muller cells, which arrest rod differentiation by releasing leukaemia inhibitory factor (LIF). These findings identify an inhibitory cell-cell interaction, which may help to explain the long delay that can occur both in vitro and in vivo between cell-cycle withdrawal and rhodopsin expression during rod development. PMID- 9199362 TI - Disruption of gastrulation and oral-aboral ectoderm differentiation in the Lytechinus pictus embryo by a dominant/negative PDGF receptor. AB - Little is known about the cell signaling involved in forming the body plan of the sea urchin embryo. Previous work suggested that PDGF-like and EGF-like receptor mediated signaling pathways are involved in gastrulation and spiculogenesis in the Lytechinus pictus embryo. Here we show that expression of the human PDGF receptor-beta lacking the cytoplasmic domain disrupted development in a manner consistent with a dominant/negative mechanism. The truncated PDGF receptor-beta inhibited gut and spicule formation and differentiation along the oral-aboral axis. The most severely affected embryos arrested at a developmental stage resembling mesenchyme blastula. Coinjection into eggs of RNA encoding the entire human PDGF receptor-beta rescued development. The truncated PDGF receptor-beta caused the aboral ectoderm-specific genes LpS1 and LpC2 to be repressed while an oral ectoderm-specific gene, Ecto-V, was expressed in all ectoderm cells. The results support the hypothesis that a PDGF-like signaling pathway plays a key role in the intercellular communication required for gastrulation and spiculogenesis, and in cell commitment and differentiation along the oral-aboral axis. PMID- 9199363 TI - The Drosophila kinesin-like protein KLP3A is required for proper behavior of male and female pronuclei at fertilization. AB - Drosophila melanogaster females homozygous for mutations in the gene encoding the kinesin-like protein KLP3A are sterile (Williams et al., 1995). We have investigated the basis of this sterility. The eggs produced by KLP3A mutant mothers are fertilized by sperm, and female meiosis appears to occur normally. However, the large majority of these embryos arrest their development soon thereafter with a characteristic phenotype. The four nuclei produced by female meiosis associate together in a polar body-like structure, while a bipolar spindle is established around the metaphase-arrested male pronucleus. Thus, the major defect caused by depletion of the KLP3A protein is either in specification of the female pronucleus, or in migration of the male and female pronuclei toward each other. We have also found that the KLP3A protein is located throughout the metaphase spindle during meiosis and the early embryonic mitotic divisions, but later accumulates specifically at the midzone of these same spindles during telophase. The protein is also present on two other microtubule structures: the sperm aster; and the radial, monastral array of microtubules established between the two meiosis II spindles. We discuss these results in light of possible functions of the KLP3A protein in pronuclear specification and migration. PMID- 9199364 TI - Melanocyte development in vivo and in neural crest cell cultures: crucial dependence on the Mitf basic-helix-loop-helix-zipper transcription factor. AB - The more than 20 different Mitf mutations in the mouse are all associated with deficiencies in neural crest-derived melanocytes that range from minor functional disturbances with some alleles to complete absence of mature melanocytes with others. In the trunk region of wild-type embryos, Mitf-expressing cells that coexpressed the melanoblast marker Dct and the tyrosine kinase receptor Kit were found in the dorsolateral neural crest migration pathway. In contrast, in embryos homozygous for an Mitf allele encoding a non-functional Mitf protein, Mitf expressing cells were extremely rare, no Dct expression was ever found, and the number of Kit-expressing cells was much reduced. Wild-type neural crest cell cultures rapidly gave rise to cells that expressed Mitf and coexpressed Kit and Dct. With time in culture, Kit expression was increased, and pigmented, dendritic cells developed. Addition of the Kit ligand Mgf or endothelin 3 or a combination of these factors all rapidly increased the number of Dct-positive cells. Cultures from Mitf mutant embryos initially displayed Mitf-positive cells similar in numbers and Kit-expression as did wild-type cultures. However, Kit expression did not increase with time in culture and the mutant cells never responded to Mgf or endothelin 3, did not express Dct, and never showed pigment. In fact, even Mitf expression was rapidly lost. The results suggest that Mitf first plays a role in promoting the transition of precursor cells to melanoblasts and subsequently, by influencing Kit expression, melanoblast survival. PMID- 9199365 TI - Enhanced cardiogenesis in embryonic stem cells overexpressing the GATA-4 transcription factor. AB - GATA-4 is a cardiac-specific member of the GATA family of zinc finger transcription factors. During embryogenesis, GATA-4 expression is detected very early in the cardiogenic area and persists later in the developing heart. Studies have shown that GATA-4 is a potent transcriptional activator of several cardiac muscle-specific genes and a key regulator of the cardiomyocyte gene program. Consistent with a role for GATA-4 in cardiomyocyte formation, inhibition of GATA 4 expression by antisense transcripts interferes with expression of cardiac muscle genes and blocks development of beating cardiomyocytes in P19 embryonic stem cells. In order to better define the function of GATA-4 in cardiogenesis, we have carried out molecular analysis of early stages of cardiomyocyte differentiation in GATA-4-deficient P19 cell lines and in P19 cells stably overexpressing GATA-4. The results indicate that GATA-4 is not required for either endodermal or mesodermal commitment or for initiation of the cardiac pathway. However, in the absence of GATA-4, differentiation is blocked at the precardiac (cardioblasts) stage and cells are lost through extensive apoptosis. In contrast, ectopic expression of GATA-4 in P19 cells accelerates cardiogenesis and markedly increases (over 10-fold) the number of terminally differentiated beating cardiomyocytes following cell aggregation. Together, these findings suggest that, in addition to its role in activation of the cardiac genetic program, GATA-4 may be the nuclear target of inductive and/or survival factors for precardiac cells. PMID- 9199366 TI - The Pax protein Noi is required for commissural axon pathway formation in the rostral forebrain. AB - No-isthmus (Noi) is a member of the zebrafish Pax family of transcriptional regulators that is expressed in restricted domains of the developing CNS. In the developing eye and optic nerve, the Noi+ cells are primitive glial cells that line the choroid fissure and optic stalk/nerve to its junction with the optic tract. This pattern of Noi expression is retained in the adult, defining the optic nerve astroglia, which wrap the left and right nerves separately at the midline, thus forming the bodily crossed optic chiasm found in fish. In embryos carrying mutations in the noi gene, the choroid fissure fails to close, glial cells of the optic nerve fail to differentiate and optic axons exhibit abnormal trajectories exiting the eye and at the midline of the diencephalon. Optic axons select inappropriate pathways into the contralateral optic nerve, rostrally towards the anterior commissure and along the ipsilateral optic tract. Noi+ cells also border the pathway of axons in the postoptic commissure, which is located adjacent to the optic chiasm. These postoptic commissural axons are defasciculated and also exhibit pathfinding defects in noi- embryos. These results indicate that Noi is required in cells that line the pathways taken by optic and non-optic commissural axons for guidance across the midline of the diencephalon. We find that expression of two members of the Netrin family of axon guidance molecules and the signalling protein Sonic hedgehog is disturbed in noi- embryos, whereas several members of the Eph family of receptors and ligands show no obvious alterations in expression at the diencephalic midline. PMID- 9199367 TI - The microtubule motor cytoplasmic dynein is required for spindle orientation during germline cell divisions and oocyte differentiation in Drosophila. AB - During animal development cellular differentiation is often preceded by an asymmetric cell division whose polarity is determined by the orientation of the mitotic spindle. In the fruit fly, Drosophila melanogaster, the oocyte differentiates in a 16-cell syncytium that arises from a cystoblast which undergoes 4 synchronous divisions with incomplete cytokinesis. During these divisions, spindle orientation is highly ordered and is thought to impart a polarity to the cyst that is necessary for the subsequent differentiation of the oocyte. Using mutations in the Drosophila cytoplasmic dynein heavy chain gene, Dhc64C, we show that cytoplasmic dynein is required at two stages of oogenesis. Early in oogenesis, dynein mutations disrupt spindle orientation in dividing cysts and block oocyte determination. The localization of dynein in mitotic cysts suggests spindle orientation is mediated by the microtubule motor cytoplasmic dynein. Later in oogenesis, dynein function is necessary for proper differentiation, but does not appear to participate in morphogen localization within the oocyte. These results provide evidence for a novel developmental role for the cytoplasmic dynein motor in cellular determination and differentiation. PMID- 9199368 TI - Essential role of heparan sulfates in axon navigation and targeting in the developing visual system. AB - Heparan sulfate (HS) is abundant in the developing brain and is a required co factor for many types of fibroblast growth factor (FGF) signaling in vitro. We report that some HSs, when added exogenously to the developing Xenopus optic pathway, severely disrupt target recognition causing axons from the retina to bypass their primary target, the optic tectum. Significantly, HS sidechains from a neuroepithelial perlecan variant that preferentially bind FGF-2, HS(FGF-2), cause aberrant targeting, whereas those that preferentially bind FGF-1 do not. Charge-matched fragments of HS(FGF-2) show that the mistargeting activity associates with the FGF-binding fragments. Heparitinase removal of native HSs at the beginning of optic tract formation retards retinal axon elongation; addition of FGF-2 restores axon extension but axons lose directionality. Late HS removal, after axons have extended through the tract, elicits a tectal bypass phenotype indicating a growth promoting and guidance function for native HSs. Our results demonstrate that different HS sidechains from the same core protein differentially affect axon growth in vivo, possibly due to their distinct FGF binding preferences, and suggest that growth factors and HSs are important partners in regulating axon growth and guidance in the developing visual system. PMID- 9199369 TI - Analysis of growth factor and receptor mRNA levels during development of the rat seminal vesicle and prostate. AB - Development of the mammalian male accessory sexual organs requires both androgens and mesenchymal/epithelial interactions. Paracrine acting factors whose expression is mesenchymal and androgen dependent have been proposed to regulate development of these organs, although the identity of these paracrine mediators is unknown. Keratinocyte growth factor (Kgf) has been shown to play an important role in the development of the mouse seminal vesicle and rat ventral prostate. Also, Kgf is expressed in mesenchymal cells and has been shown to be regulated by androgens in prostatic cells grown in vitro. Thus Kgf has been proposed as a mediator of androgen action. We have investigated the expression of Kgf mRNA during development of the rat seminal vesicle and prostate, both in vitro and in vivo. Additionally we have examined mRNAs for Kgf receptor (KgfR), transforming growth factor alpha (Tgf alpha), epidermal growth factor receptor (EgfR) and cytokeratin 19 (CK19). The levels of growth factor and receptor mRNAs fluctuated during androgen-regulated development; however, these changes reflected variations in the mesenchymal/epithelial ratio rather than regulation by testosterone. Expression of Kgf is mesenchymal, while KgfR is epithelial and Tgf alpha is predominantly epithelial. The changes in the levels of mRNAs for these factors correlated well with changes in the level of an epithelial marker, CK19, suggesting they were due to alterations in the relative abundance of tissue compartments in which they were expressed. Kgf has been shown to mimic androgen action in explant cultures of seminal vesicle and prostate. We demonstrate here that anti-androgens are able to block Kgf stimulated development, suggesting that Kgf and androgen receptor signalling pathways may interact. Taken together our data suggest that, in vivo, Kgf may interact with androgen receptor signalling but it is not a direct target of androgen action. PMID- 9199370 TI - Clonal dispersion and evidence for asymmetric cell division in ferret cortex. AB - Cell lineage analysis with retroviral libraries suggests that clonal progeny disperse widely in rodent cortex. To determine whether widespread dispersion is a general mammalian plan and to investigate phylogenetic differences in cortical development, we analyzed cell lineage in the ferret, a carnivore and near relative of the cat. The ferret possesses a highly developed, folded cerebral cortex, characteristic of higher mammalian species. Progenitor cells of the ferret cerebral cortex were tagged with an amphotropic retroviral library encoding alkaline phosphatase, and sibling relationships were determined using the polymerase chain reaction. Neuronal clones were single neurons (52%) or large clones (48%; average, 7 neurons) containing neurons and glia in widespread cortical locations. Neuronal clones in the ferret labeled at middle to late neurogenesis (embryonic day 33-35) contained large numbers of neurons and showed little tendency to cluster. The large proportion of single neuron clones, contrasted with the large size of multicell clones, suggests that some progenitors divide asymmetrically, producing a postmitotic neuron and regenerating a multipotential cell. PMID- 9199371 TI - Involvement of programmed cell death in morphogenesis of the vertebrate inner ear. AB - An outstanding challenge in developmental biology is to reveal the mechanisms underlying the morphogenesis of complex organs. A striking example is the developing inner ear of the vertebrate, which acquires a precise three dimensional arrangement of its constituent epithelial cells to form three semicircular canals, a central vestibule and a coiled cochlea (in mammals). In generating a semicircular canal, epithelial cells seem to 'disappear' from the center of each canal. This phenomenon has been variously explained as (i) transdifferentiation of epithelium into mesenchyme, (ii) absorption of cells into the expanding canal or (iii) programmed cell death. In this study, an in situ DNA end labeling technique (the TUNEL protocol) was used to map regions of cell death during inner ear morphogenesis in the chicken embryo from embryonic days 3.5-10. Regions of cell death previously identified in vertebrate ears have been confirmed, including the ventromedial otic vesicle, the base of the endolymphatic duct and the fusion plates of the semicircular canals. New regions of cell death are also described in and around the sensory organs. Reducing normal death using retrovirus-mediated overexpression of human bcl-2 causes abnormalities in ear morphogenesis: hollowing of the center of each canal is either delayed or fails entirely. These data provide new evidence to explain the role of cell death in morphogenesis of the semicircular canals. PMID- 9199372 TI - A novel group of pumilio mutations affects the asymmetric division of germline stem cells in the Drosophila ovary. AB - Germline stem cells play a pivotal role in gametogenesis; yet little is known about how they are formed, how they divide to self-renew, and how these processes are genetically controlled. Here we describe the self-renewing asymmetric division of germline stem cells in the Drosophila ovarian germline, as marked by the spectrosome, a cytoplasmic structure rich in membrane skeletal proteins. The ontogeny of the spectrosome marks the lineage of germline stem cells. We identified two new groups of mutations in which the divisional asymmetry is disrupted. The first, which we refer to as ovarette (ovt) mutations, was shown to correspond to a novel class of mutations in the pumilio locus. Since pumilio is known to posttranscriptionally repress the expression of target genes at earlier stages of germ cell development, our results suggest that a similar activity is needed to maintain germ line stem cells. We have also identified a second and novel gene, piwi, whose mutations abolish germline stem cell division. PMID- 9199373 TI - Mechanisms of dorsal-ventral patterning in noggin-induced neural tissue. AB - We have investigated mechanisms of dorsal-ventral patterning of neural tissue, using Xenopus ectoderm neuralized by noggin protein. This tissue appears to be patterned dorsoventrally; cp1-1, a gene expressed in the dorsal brain, and etr-1, a gene largely excluded from the dorsal brain, are expressed in separate territories in noggin-treated explants (Knecht, A. K., Good, P. J., Dawid, I. B. and Harland, R. M. (1995) Development 121, 1927-1936). Here we show further evidence that this pattern represents a partial dorsal-ventral organization. Additionally, we test two mechanisms that could account for this pattern: a dose dependent response to a gradient of noggin protein within the explant, and regulative cell-cell interactions. We show that noggin exhibits concentration dependent effects, inducing cp1-1 at low doses but repressing it at high doses. Since noggin acts by antagonizing Bone Morphogenetic Protein (BMP) signaling, this result suggests that BMPs also may act in a dose-dependent manner in vivo. However, in the absence of a noggin gradient, regulative cell-cell interactions can also pattern the tissue. Such regulation is facilitated by increased motility of noggin-treated cells. Finally, the response of cells to both of these patterning mechanisms is ultimately controlled by a third process, the changing competence of the responding tissue. PMID- 9199374 TI - Pollen tube guidance by the female gametophyte. AB - In flowering plants, pollen grains germinate on the pistil and send pollen tubes down the transmitting tract toward ovules. Previous genetic studies suggested that the ovule is responsible for long-range pollen tube guidance during the last phase of a pollen tube's journey to the female gametes. It was not possible, however, to unambiguously identify the signaling cells within an ovule: the haploid female gametophyte or the diploid sporophytic cells. In an effort to distinguish genetically between these two possibilities, we have used a reciprocal chromosomal translocation to generate flowers wherein approximately half the ovules do not contain a functional female gametophyte but all ovules contain genotypically normal sporophytic cells. In these flowers, pollen tubes are guided to the normal but not to the abnormal female gametophytes. These results strongly suggest that the female gametophyte is responsible for pollen tube guidance, but leave open the possibility that the gametophyte may accomplish this indirectly through its influence on some sporophytic cells. PMID- 9199375 TI - Adult respiratory distress syndrome, pneumonia, and mortality following thoracic injury and a femoral fracture treated either with intramedullary nailing with reaming or with a plate. A comparative study. AB - Multiply injured patients (an Injury Severity Score of 17 points or more) who were admitted to one of two level-I regional trauma centers between 1983 and 1994 because of a fracture of the femoral shaft with a thoracic injury (an Abbreviated Injury Scale score of 2 points or more) or without a thoracic injury were studied retrospectively. The patient populations and the protocols for the treatment of trauma were similar at the two centers; however, the centers differed with regard to the technique that was used for acute stabilization of the fracture of the femoral shaft. At Center I intramedullary nailing with reaming was used in 217 (95 per cent) of the 229 patients, whereas at Center II a plate was used in 206 (92 per cent) of the 224 patients. This difference was used to investigate the effect of acute femoral reaming on the occurrence of adult respiratory distress syndrome in multiply injured patients who had a chest injury. Three groups of patients were evaluated: those who had both a fracture of the femur and a thoracic injury, those who had a fracture of the femur but no thoracic injury, and those who had a thoracic injury without a fracture of the femur or the tibia. The third group was studied at each center to determine if there was a difference between the institutions with regard to the rate of adult respiratory distress syndrome. Patients who had diabetes, chronic obstructive pulmonary disease, asthma, hepatic or renal failure, or an immunosuppressive condition were excluded from the study. The records were abstracted to determine the Injury Severity Score, Abbreviated Injury Scale score, and Glasgow Coma Score for each patient. Requirements for fluid resuscitation were calculated for the first twenty-four hours; these included the number of units of packed red blood cells, fresh-frozen plasma, and platelets that were transfused and the volume of crystalloid that was used. The duration of intubation, the duration of hospitalization, and the occurence of adverse outcomes (death, multiple organ failure, adult respiratory distress syndrome, pneumonia, and pulmonary embolism) were determined for each patient. The groups of patients were analyzed as a whole and then were stratified into subgroups (according to whether or not they had a thoracic injury and whether the Injury Severity Score was less than 30 points or 30 points or more) to determine if the type of fixation of the femoral fracture affected the rate of adult respiratory distress syndrome or mortality. Logistic regression models were used to analyze the data. The over-all occurrence of adult respiratory distress syndrome in the 453 patients who had a femoral fracture was only 2 per cent (ten patients). The rates of adult respiratory distress syndrome for the patients who had a thoracic injury but no femoral fracture (eight [6 per cent] of 129 patients at Center I, compared with ten [8 per cent] of 125 patients at Center II) did not differ between centers, suggesting that the institutions were comparable in their treatment of multiply injured patients. The occurrence of adult respiratory distress syndrome in the patients who had a femoral fracture without a thoracic injury did not differ substantially according to whether the fracture had been treated with a nail (118 patients) or a plate (114 patients). Likewise, the frequency of adult respiratory distress syndrome, pneumonia, pulmonary embolism, failure of multiple organs, or death for the patients who had a femoral fracture and a thoracic injury was similar regardless of whether nailing with reaming (117 patients) or a plate (104 patients) had been used. The use of intramedullary nailing with reaming for acute stabilization of fractures of the femur in multiply injured patients who have a thoracic injury without a major comorbid disease does not appear to increase the occurrence of adult respiratory distress syndrome, pulmonary embolism, failure of multiple organs, pneumonia, or death. PMID- 9199376 TI - Long-term outcome after open reduction through an anteromedial approach for congenital dislocation of the hip. AB - We reviewed the long-term outcome of open reduction of ninety-three congenitally dislocated hips (in seventy-six children) through an anteromedial approach. The average age of the patients was fourteen months (range, two to fifty months) at the time of the reduction and eleven years (range, four to twenty-three years) at the time of the most recent follow-up evaluation. At the most recent follow-up evaluation, sixty-six hips (71 per cent) had an excellent or good result, twenty four (26 per cent) had a fair result, and three (3 per cent) had a poor result, according to the Severin classification system. An inverted neolimbus at the time of the operation and postoperative growth disturbance of the femoral head were associated with a poor roentgenographic result. According to the classification of Bucholz and Ogden, twenty-two hips (24 per cent) had type-II avascular necrosis, thirteen hips (14 per cent) had type-III, three (3 per cent) had type IV, two (2 per cent) had non-classifiable lesions, and fifty-three (57 per cent) did not have avascular necrosis. A high hip dislocation and an operation after the age of twenty-four months were associated with a higher rate of growth disturbances of the femoral head. With the numbers available for study, we did not find any association between short-term preoperative traction, ligation of the medial circumflex vessel, or the type of neolimbus and the prevalence of growth disturbances. Two hips redislocated postoperatively, and seven had transient stiffness. We consider the anteromedial approach to be useful in the management of patients with congenital dislocation of the hip who are twenty-four months old or less. The advantages of this approach include direct access to the obstacles to reduction, avoidance of damage of the iliac apophysis and the abductor muscles, minimum blood loss, the need for only a single operative session for treatment of both hips, and a cosmetically acceptable scar. The prevalence of type-II growth disturbances of the femoral head was higher than had been expected, emphasizing the need for additional investigation. PMID- 9199377 TI - Repair of the defect in spondylolysis. Durable fixation with pedicle screws and laminar hooks. AB - Direct repair of a defect in the pars interarticularis was performed with use of bone-grafting and internal fixation with a pedicle screw, rod, and laminar hook in order to achieve a higher prevalence of osseous union than that achieved with commonly used procedures. The configuration of the head of the screw, which is designed to allow it to connect with the rod at the necessary angle, simplified the placement of the rod. The procedure was performed in sixteen patients who had a bilateral defect of the pars interarticularis with or without grade-I or II spondylolisthesis, had had failure of non-operative treatment, and had had temporary relief of pain after the area of the defect in the pars interarticularis had been infiltrated with lidocaine. Concomitant degeneration of a disc was not a criterion for exclusion. The patients were followed for an average of twenty-five months (range, twenty-four to twenty-eight months). The average age at the time of the operation was thirty-two years (range, twelve to sixty years). Six patients had findings of nerve-root compression on myelography with computerized tomographic scanning, and the bone spurs overlying the affected nerve root around the defect in the pars interarticularis were removed with an ultrasonic osteotome through a small window. The implant was removed about one year after the operation. Oblique radiographs showed osseous union in the previous defect bilaterally in all sixteen patients. Thirteen patients were free of symptoms, and three had major improvement with occasional low-back pain. None had a complication, such as infection, breakage of the implant, or irritation of a nerve root. The method used for direct repair of the defect of the pars interarticularis in these patients proved to be simple and effective. Relief of symptoms appeared to depend on decompression of the affected nerve root, if one was involved, and on preoperative prediction of the locus of the symptoms by infiltration of the pars interarticularis with lidocaine. PMID- 9199378 TI - Total elbow arthroplasty as primary treatment for distal humeral fractures in elderly patients. AB - We retrospectively reviewed the results of primary total elbow arthroplasty for the treatment of an acute fracture of the distal aspect of the humerus in twenty consecutive patients (twenty-one elbows) who had a mean age of seventy-two years (range, forty-eight to ninety-two years) at the time of the injury. The patients were managed between November 1982 and October 1992. The presence of rheumatoid arthritis in nine patients (ten elbows) influenced the choice of treatment. The mean interval between the injury and the total elbow arthroplasty was seven days (range, one to twenty-five days). The mean duration of postoperative hospitalization was seven days (range, four to thirteen days). The mean duration of follow-up was 3.3 years (range, three months to 10.5 years). All patients were followed for a minimum of two years or until the time of death; the duration of follow-up was less than two years for three patients who died. None of the patients were lost to follow-up. Twenty implants were intact at the latest follow up examination. One patient had a revision total elbow arthroplasty twenty months after the index procedure because of a fracture of the ulnar component sustained in a fall on the outstretched arm. On the basis of the Mayo elbow performance score, fifteen elbows had an excellent result and five had a good result; there were inadequate data for one elbow. There were no fair or poor results. The mean arc of flexion was 25 to 130 degrees. There was no evidence of loosening on the radiographs. Postoperative complications included fracture of the ulnar component in one patient, ulnar neurapraxia in three, and reflex sympathetic dystrophy in one. The results suggest that total elbow arthroplasty can be an alternative form of treatment of a severely comminuted fracture of the distal aspect of the humerus in older patients even in the presence of rheumatoid arthritis. This procedure is not an alternative to osteosynthesis in younger patients. PMID- 9199379 TI - Quantification of the radial torsion angle with computerized tomography in cadaver specimens. AB - Torsion of a long bone is the twist along its longitudinal axis; torsion of the radius is defined by the angle between the proximal and distal metaphyses in the transverse plane. Measurement of the radial torsion angle provides a means of detection and quantification of malrotation after a fracture. The purpose of the current study was to develop and standardize a technique for the measurement of torsion of the radius. Axial computerized tomographic images of thirty-nine pairs of dry cadaver specimens of normal radii, and an additional four pairs of radii with a unilateral deformity of the distal metaphysis that was consistent with a previous fracture, were studied and a measurement protocol was established. The radial torsion angle was measured by three independent observers on two separate occasions. Reproducibility of the technique was determined with use of the intraclass correlation coefficient to express both interobserver and intraobserver reliability. Consistency of measurements between observers and by the same observer was high, with intraclass correlation coefficients ranging from 0.87 to 0.94. The mean torsion angle for the eighty-two normal radii in the study was 32.6 degrees (95 per cent confidence interval of the mean, 30.3 to 34.9 degrees; range, 1.4 to 58.8 degrees). There were small variations in torsion angle between the two radii of each normal pair (mean side-to-side difference, 4.9 degrees; 95 per cent confidence interval of the mean, 3.5 to 6.3 degrees). The mean torsion angle of the four radii with a malunited fracture was 10.4 degrees (95 per cent confidence interval of the mean, 5.7 to 15.1 degrees), and the mean side-to-side difference in the pairs containing these radii was 24.1 degrees (95 per cent confidence interval of the mean, 8.5 to 39.6 degrees; p < 0.0001 compared with the normal radii). PMID- 9199380 TI - Near-infrared spectroscopy for monitoring of tissue oxygenation of exercising skeletal muscle in a chronic compartment syndrome model. AB - Variations in the levels of muscle hemoglobin and of myoglobin oxygen saturation can be detected non-invasively with near-infrared spectroscopy. This technique could be applied to the diagnosis of chronic compartment syndrome, in which invasive testing has shown increased intramuscular pressure associated with ischemia and pain during exercise. We simulated chronic compartment syndrome in ten healthy subjects (seven men and three women) by applying external compression, through a wide inflatable cuff, to increase the intramuscular pressure in the anterior compartment of the leg. The tissue oxygenation of the tibialis anterior muscle was measured with near-infrared spectroscopy during gradual inflation of the cuff to a pressure of forty millimeters of mercury (5.33 kilopascals) during fourteen minutes of cyclic isokinetic dorsiflexion and plantar flexion of the ankle. The subjects exercised with and without external compression. The data on tissue oxygenation for each subject then were normalized to a scale of 100 per cent (the baseline value, or the value at rest) to 0 per cent (the physiological minimum, or the level of oxygenation achieved by exercise to exhaustion during arterial occlusion of the lower extremity). With external compression, tissue oxygenation declined at a rate of 1.4 +/- 0.3 per cent per minute (mean and standard error) during exercise. After an initial decrease at the onset, tissue oxygenation did not decline during exercise without compression. The recovery of tissue oxygenation after exercise was twice as slow with compression (2.5 +/- 0.6 minutes) than it was without the use of compression (1.3 +/- 0.2 minutes). PMID- 9199381 TI - Intramuscular deoxygenation during exercise in patients who have chronic anterior compartment syndrome of the leg. AB - Currently, the definitive diagnosis of chronic compartment syndrome is based on invasive measurements of intracompartmental pressure. We measured the intramuscular pressure and the relative oxygenation in the anterior compartment of the leg in eighteen patients who were suspected of having chronic compartment syndrome as well as in ten control subjects before, during, and after exercise. Chronic compartment syndrome was considered to be present if the intramuscular pressure was at least fifteen millimeters of mercury (2.00 kilopascals) before exercise, at least thirty millimeters of mercury (4.00 kilopascals) one minute after exercise, or at least twenty millimeters of mercury (2.67 kilopascals) five minutes after exercise. Changes in relative oxygenation were measured with use of the non-invasive method of near-infrared spectroscopy. In all patients and subjects, there was rapid relative deoxygenation after the initiation of exercise, the level of oxygenation remained relatively stable during continued exercise, and there was reoxygenation to a level that exceeded the pre-exercise resting level after the cessation of exercise. During exercise, maximum relative deoxygenation in the patients who had chronic compartment syndrome (mean relative deoxygenation [and standard error], -290 +/- 39 millivolts) was significantly greater than that in the patients who did not have chronic compartment syndrome ( 190 +/- 10 millivolts) and that in the control subjects (-179 +/- 14 millivolts) (p < 0.05 for both comparisons). In addition, the interval between the cessation of exercise and the recovery of the pre-exercise resting level of oxygenation was significantly longer for the patients who had chronic compartment syndrome (184 +/- 54 seconds) than for the patients who did not have chronic compartment syndrome (39 +/- 19 seconds) and the control subjects (33 +/- 10 seconds) (p < 0.05 for both comparisons). PMID- 9199382 TI - Bankart repair for anterior instability of the shoulder. Long-term outcome. AB - Anterior instability of the shoulder is a commonly encountered entity in orthopaedic practice. The Bankart procedure is considered by many surgeons to be the treatment of choice for this condition. Despite its widespread popularity, there have been no studies on the long-term outcome of the Bankart procedure as far as we know. Sixty shoulders (fifty-six patients) that had been followed for a minimum of eight years after a Bankart procedure were evaluated for range of motion, stability, and strength according to the data form of the American Shoulder and Elbow Surgeons for examination of the shoulder. The results for the involved shoulder were compared with the findings for the contralateral, normal shoulder. All patients completed a questionnaire regarding the history of the instability of the shoulder, the level of participation in sports before and after the operation, the preoperative and postoperative level of pain, and whether the patient had ever sustained a dislocation that needed reduction by a physician. Information about the current ability of the patient to function at home, at work, and during sports also was requested. In addition, the patients were asked to rate the results of the operation and to indicate whether they would have the same procedure again for the same problem. At a mean of 11.9 years after the operation, the mean loss of external rotation was 12 degrees (range, 0 to 30 degrees) (p < 0.0001). There were no significant differences in forward elevation, abduction, or internal rotation between the involved shoulder and the contralateral, normal shoulder. One patient had crepitus on glenohumeral motion. Fifty-five of the fifty-six patients returned to the occupation that they had had preoperatively, without having to alter their activities. Twenty-eight patients had mild pain with strenuous activity, and one patient had pain at rest. Three patients had a dislocation of the involved shoulder because of a new traumatic event more than three years postoperatively. Fifty-two patients rated the result as good or excellent; three, as fair; and one, as poor. Fifty-four patients said that they would have a Bankart procedure performed again for the same problem. We present a new system for rating the shoulder that emphasizes function and is based specifically on the goals stated by the patients to be most important with regard to the shoulder. Using this system, we found that the Bankart procedure offers an excellent objective long-term outcome with a high degree of patient satisfaction. PMID- 9199383 TI - Demographic biases of scoring instruments for the results of total knee arthroplasty. AB - Four knee-scoring systems were used to evaluate 200 adult subjects who had no history of injury, abnormality, or treatment of the knees, hips, lower extremities, or spine. All subjects were in the age-range (fifty to 100 years; average, 65.5 years) typical of candidates for total knee replacement. In addition to a physical examination, complete demographic data were collected for each subject. The knee scores were normalized by dividing the observed score by the maximum possible score. The average normalized total knee score was 91 per cent (range, 22 to 100 per cent) according to the knee score of The Hospital for Special Surgery, 95 per cent (range, 10 to 100 per cent) according to the system of Hungerford and Kenna, 89 per cent (range, -7.75 to 100 per cent) according to a modification of the scoring system of The Knee Society, and 95 per cent (range, 26.5 to 100 per cent) according to the system of Hofmann et al. Demographic variables that had a significant negative correlation with the knee scores included advanced age (particularly of eighty-five years or more), a family income below the poverty level, and two major medical conditions or more. Observed differences in knee scores between different study groups that have not been matched for various clinically relevant factors are at least as likely to represent differences in the patient populations as they are to represent differences in the operative technique or the design of the implant. PMID- 9199384 TI - Medial gastrocnemius transposition flap for the treatment of disruption of the extensor mechanism after total knee arthroplasty. AB - We describe a modified technique for the salvage of a total knee arthroplasty after disruption of the extensor mechanism. Between January and December 1992, seven patients had reconstruction of the extensor mechanism with use of a medial or an extended medial gastrocnemius flap. Six of the seven patients were followed for a mean of thirty-three months (range, twenty-six to forty-one months) and were evaluated both preoperatively and postoperatively with regard to the knee and functional scores of The Knee Society as well as the range of motion, extensor lag, walking status, and patellar height. The seventh patient was lost to follow-up six months postoperatively and was excluded from the analysis of the results. Preoperatively, the knee and functional scores were 16 +/- 12.3 points and 12 +/- 12.1 points (mean and standard deviation), respectively; the mean range of motion was 70 +/- 44.0 degrees; and the mean extensor lag was 53 +/- 33.4 degrees. Postoperatively, the mean knee and functional scores improved to 82 +/- 12.4 points and 51 +/- 23.0 points, respectively; the mean range of motion improved to 100 +/- 21.8 degrees; and the mean extensor lag decreased to 24 +/- 18.8 degrees. After the procedure, all patients who previously had been dependent on a walker were able to walk about the community with or without a cane, and those who had been dependent on a wheelchair were able to walk with the assistance of a walker. Patellar height was measured according to the method of Insall and Salvati for the four patients who had a patella. Preoperatively, the patellar heights were grossly abnormal; postoperatively, they more closely approached accepted normal values for three of the four patients. Reconstruction of a complicated rupture of the extensor mechanism with use of a medial gastrocnemius transposition flap after total knee arthroplasty is a reliable option for treatment. PMID- 9199385 TI - Pyogenic vertebral osteomyelitis. AB - I retrospectively reviewed the records of 111 patients who had pyogenic vertebral osteomyelitis unrelated to an open procedure on the spine. The mean age at the time of the diagnosis was sixty years (range, eighteen to eighty-four years); sixty-one patients (55 per cent) were sixty years old or more. Forty-four patients (40 per cent) had an impaired immune system secondary to diabetes mellitus, the use of corticosteroids, chemotherapy for cancer, rheumatic or immunological disease, renal or hepatic failure, malnutrition, or myelodysplasia. Magnetic resonance imaging, critical for the determination of an early diagnosis, was performed for 103 patients (93 per cent). The infection in sixty-eight patients (61 per cent) was diagnosed within one month after the onset of symptoms. The most frequent infecting organism was Staphylococcus aureus (forty patients; 36 per cent). The infection in forty-one patients (37 per cent) was caused by organisms, such as Staphylococcus epidermidis, Propionibacterium acnes, and diphtheroid species, that are traditionally considered to be of low virulence. The urinary tract was the most frequent source of infection (confirmed in thirteen patients and suspected in twenty-one). The success of non-operative treatment was predicted by four independent variables: an age of less than sixty years, the immune status, infection with Staphylococcus aureus, and a decreasing erythrocyte sedimentation rate. Forty-two patients were managed with debridement and arthrodesis. Fourteen of these patients also had instrumentation of the spine, in the presence of infection, without compromise of the outcome. Eighteen patients died by the time of the latest follow-up evaluation at a mean of four years (range, two years and two months to six years and six months): seven who had been managed non-operatively died in the first month after the diagnosis was made, three died in the acute postoperative period, three died of late complications of paraplegia, and five died of unrelated causes. None of the eighty-nine patients who were seen at a minimum of two years postoperatively had had late recurrence of infection. Chronic, severe back pain was noted in only seven patients. PMID- 9199386 TI - Treatment of complications of shoulder arthrodesis. AB - A reconstructive osteotomy was performed to correct symptomatic malposition after arthrodesis of the shoulder in nine of fourteen patients who had complications related to the arthrodesis. The clinical position of the arm in relation to the trunk was determined with the method described by Rowe. Malposition was primarily the result of fusion in more than 15 degrees of either flexion or abduction, or both, coupled with improper rotation, defined as rotation of less than 40 degrees or more than 60 degrees. Reconstructive osteotomy eliminated pain and improved the ability of the patient to perform six activities of daily living. The complications necessitating operative treatment after the arthrodesis in the remaining five patients included failure of the arthrodesis site to unite (three patients), a wound hematoma at the iliac-crest donor site (one patient), and a superficial wound infection (one patient). Two additional complications - a fracture through a screw-hole in the humerus and a fracture distal to the internal fixation device - occurred after the reconstructive osteotomies for malposition. All of the complications resolved with treatment. Arthrodesis of the shoulder is a technically demanding procedure that can lead to serious complications that necessitate operative intervention. Careful attention to operative technique and to the position of the arthrodesis are essential. PMID- 9199387 TI - Oncological outcomes of operative treatment of subcutaneous soft-tissue sarcomas of the extremities. AB - We reviewed the cases of sixty-two patients who had had a subcutaneous sarcoma to determine the effect of tumor and treatment-related variables on the rates of survival and local recurrence. Fifty-nine (95 per cent) of the patients had had an operation at another hospital before being referred to us. Twenty-nine (47 per cent) of the sixty-two tumors were high-grade, forty-two (68 per cent) were small (five centimeters or less), and thirty (48 per cent) were malignant fibrous histiocytomas. We followed a treatment strategy that consisted of repeat excision with the goal of obtaining wide margins. Excluding thirteen patients who had had a palpable local recurrence at the time of presentation, twenty (49 per cent) of forty-one patients who had had a marginal excision at another hospital had microscopic residual tumor on repeat excision. At a median of fifty-six months after the repeat excision, fifty (81 per cent) of the sixty-two patients had been continuously disease-free, one had no evidence of disease, eight had died of the disease, and three had died of other causes. The five-year rate of disease-free survival was 85 per cent (fifty-three of sixty-two patients). There were three local recurrences, all in patients who had had a marginal resection. No recurrences were noted in patients who had had a wide local excision of the tumor or of the previous operative field. Multivariate analysis revealed that a large tumor (greater than five centimeters), a marginal excision, and adjuvant radiation therapy were associated with a worse prognosis. Excellent rates of survival for patients who have a subcutaneous sarcoma, including those who have a large or high-grade tumor and those who have residual tumor following a previous operation, can be obtained with carefully planned operative treatment alone. We recommend operative excision or repeat excision with wide margins because of the high prevalence of residual tumor. Size is the most important tumor-related factor, and the operative margin is the most important treatment-related factor. The additional value of adjuvant radiation therapy remains unproved. PMID- 9199388 TI - Enchondroma of the distal phalanx of the hand. AB - We saw five patients who had enchondroma of the distal phalanx, a relatively uncommon site for that lesion. Three patients had pain secondary to a pathological fracture and were managed with curettage and bone-grafting through a palmar longitudinal incision. The other two patients had severe deformities of the fingertip and nail. One was managed with disarticulation of the distal interphalangeal joint and the other, with curettage and grafting through a dorsal approach followed by reconstruction of the nail matrix. We believe that the palmar incision in the pulp of the finger has few, if any, complications. PMID- 9199389 TI - Fracture of an acetabular component inserted without cement. A case report. PMID- 9199390 TI - Myositis ossificans of the piriformis muscle: an unusual cause of piriformis syndrome. A case report. PMID- 9199391 TI - Sclerotic lesion of the tibia without involvement of lymph nodes. Report of an unusual case of Rosai-Dorfman disease. PMID- 9199392 TI - Treatment of metastatic adenocarcinoma of the pelvis and the extremities. PMID- 9199393 TI - Use of the Milwaukee brace for progressive idiopathic scoliosis. PMID- 9199394 TI - Use of the Milwaukee brace for progressive idiopathic scoliosis. PMID- 9199395 TI - Gene therapy for central nervous system injury: the use of cationic liposomes: an invited review. AB - This paper briefly reviews general principles of gene therapy with emphasis on the therapeutic potential of cationic liposome-mediated neurotrophin gene transfer to treat central nervous system (CNS) injury. Current developments in studies of gene therapy for CNS injury are both impressive and promising. Ex vivo gene transfer into the CNS is relatively mature in animal studies following more than a decade of experimental studies. In vivo gene transfer into the CNS has gained more attention recently. Although progress has been made using viral vectors, rapid advances in transfection technologies employing cationic liposomes, together with the relatively low toxicity of these nonviral vector systems, suggest that liposomes may have significant potential for clinical applications. Although many investigators have recognized that gene therapy may be useful for treatment of certain genetic defect diseases or cancer, gene therapy for CNS injury is relatively novel. In contrast to genetic defect disorders, temporary induction of transgenes may have therapeutic applications for CNS injuries such as stroke and trauma. Employing gene transfer techniques to achieve therapeutically useful levels of expression of neurotrophins in the CNS could provide a new strategy for treatment of the traumatically injured CNS. PMID- 9199396 TI - Anoxic injury in the rat spinal cord: pharmacological evidence for multiple steps in Ca(2+)-dependent injury of the dorsal columns. AB - To examine anoxic injury in spinal cord white matter, we studied axonal conduction in the dorsal columns during and following a standard 60 min anoxic insult at 36 degrees C. Perfusion of the spinal cord in 0-Ca2+ Ringer solution resulted in significantly improved recovery of the compound action potential. Similarly, removal of Na+ from the perfusate resulted in significantly improved recovery of conduction in dorsal column axons. Exposure of the anoxic spinal cord to the Na+ channel blocker tetrodotoxin (TTX), the Na-Ca exchange blockers benzamil and bepridil, Na(+)-H+ exchange blockers amiloride and harmaline, and perfusion in Ringer solution with pH adjusted to 6.4, all resulted in improved recovery. The tertiary anesthetics procaine and lidocaine, as well as phenytoin and carbamazepine, also resulted in improved recovery of compound action potential amplitude after 60 min of anoxia. These results demonstrate that a significant component of irreversible loss of conduction, following anoxic injury of the dorsal columns, is Ca(2+)-dependent. Moreover, these results demonstrate that TTX-inhibitable Na+ channels participate in the pathophysiology of anoxic injury in spinal cord white matter, and indicate that reverse Na-Ca exchange provides a route for at least part of the damaging influx of Ca2+ into an intracellular compartment in anoxic spinal cord white matter. Our results also suggest that extracellular acidosis may have a protective effect on anoxic spinal cord white matter, and support the hypothesis that anoxic injury of spinal cord white matter may involve the Na(+)-H+ exchanger. PMID- 9199397 TI - Histopathologic clues to the pathways of neuronal death following ischemia/hypoxia. AB - This review describes histopathologic observations made with both light and electron microscopy using both conventional staining techniques and histochemistry. Several conditions are analyzed: Ischemic cell change; delayed neuronal death; selective vulnerability. The histopathologic support for the calcium hypothesis and for the excitotoxic hypothesis explaining neuronal death is also reviewed. The findings lead to several suggestions relevant to attempts at developing interventional therapies administered after the onset of ischemia/hypoxia. (1) Except in gerbils, delayed neuronal death and more rapid neuronal death appear to be on the same continuum of cellular events. The lag between ischemia and either onset or termination of these shared events depends upon the severity and/or duration of ischemia/hypoxia. We still do not know whether the "delay," when it occurs, is a delay between ischemia and initiation of the lethal sequence or is, instead, a delay between an immediate initiation of the sequence and its lethal termination. (2) Selective vulnerability (e.g., of CA1 sector in hippocampus) is only relative. The changes are again those of ischemic cell change and are identical to the changes seen elsewhere in more severe ischemia. (3) There is histopathologic support for both the calcium hypothesis and for the cytotoxic hypothesis. Indeed, there is histopathologic support linking the two hypotheses and linking these mechanisms to the appearance of ischemic cell change. However, the histopathologic data are surprisingly sparse. The role of either hypothesis in explaining neuronal death in all areas of brain, in all types of ischemic insult, and at all times following such an insult remains to be established. (3) Apoptosis may be an important mode of neuronal death following ischemia. It differs from acute ischemic cell change; nevertheless, both calcium overload and/or excitotoxic neurotransmitters may trigger apoptosis. (4) Third cell change has been described: Eosinophilic neurons that are not shrunken and whose nuclei are not pyknotic but contain clumped chromatin. The pathogenesis and fate of these neurons remains uncertain. It is possible that they represent early apoptotic neurons. Adequate assessment of apoptosis and its relationship (to both these neurons and to neurons displaying classical ischemic cell change) may depend upon dual staining with conventional aniline dyes and with histochemical techniques designed to detect intranuclear fragments of DNA. PMID- 9199398 TI - Lack of delayed effects of amphetamine, methoxamine, and prazosin (adrenergic drugs) on behavioral outcome after lateral fluid percussion brain injury in the rat. AB - This study examined the delayed effects of the administration of d-amphetamine, methoxamine (an alpha1-adrenergic receptor agonist), and prazosin (an alpha1 adrenergic receptor antagonist) on the behavioral outcome of lateral fluid percussion (FP) brain injury. Rats trained to perform a beam-walking task were subjected to brain injury of moderate severity (2.1 to 2.2 atm). Twenty-four hours after injury, rats were treated with amphetamine, methoxamine, or prazosin at two or three different dose levels. Amphetamine-treated animals displayed no significant improvement in beam-walking ability either during or after drug intoxication (from days 3 to 5 after brain injury). Similarly, neither methoxamine nor prazosin significantly affected beam-walking ability during or after drug intoxication. Neither amphetamine treatment at three different doses nor treatment with methoxamine or prazosin at two different doses affected the spatial learning disabilities of brain-injured animals. These results suggest that (1) unlike amphetamine administration after sensorimotor cortex (SMC) ablation or contusion brain injury models, amphetamine administration at 24 h after concussive FP brain injury does not improve beam-walking performance; (2) unlike amphetamine administration 10 min after concussive FP brain injury amphetamine administration 24 h after injury does not improve cognitive function; and (3) unlike prazosin administration after SMC ablation brain injury, prazosin administration 24 h after concussive FP brain injury does not effect beam-walking performance. PMID- 9199399 TI - The effect of dietary alpha-tocopherol on the experimental vasogenic brain edema. AB - It has become increasingly obvious that free radicals and lipid peroxidation contribute to brain damage from trauma by mediating edema formation and ischemia. It should, therefore, be expected that the actual level of endogenous antioxidants, as for example, vitamin C and E in plasma, has an influence on the extent of free radical-induced injury. In this communication we investigate the effect of dietary changes in the free radical scavenger alpha-tocopherol on posttraumatic cerebral swelling in Sprague-Dawley rats. Low, normal, and high plasma levels of alpha-tocopherol were established by respective diets supplied over 2 weeks. Animals of all groups received the same food without alpha tocopherol. One group was fed a vitamin E-free diet. The pellet-food for the other animals was supplemented either with 5-mg alpha-tocopherol/100 g or 250-mg alpha-tocopherol/100 g dry mass, respectively. The vitamin E-free diet lowered the alpha-tocopherol level in plasma to 30% of control, whereas supplementation with 250 mg/100 g led to a plasma concentration of 200% of control. The animals were then subjected to a focal cold injury of the left cerebral hemisphere. Twenty-four hours after trauma the brain was removed and the water content of each hemisphere was determined by the wet-dry weight method. Swelling of the traumatized hemisphere was calculated as the difference in weight between the traumatized and contralateral control hemisphere. The 2-week alpha-tocopherol supplementation or -deletion diet, respectively, did not either afford significant reduction or lead to an enhancement of traumatic brain swelling. Likewise, the increase in brain water content of the traumatized hemisphere was not affected. It is concluded that supplementation or depletion of alpha tocopherol for 2 weeks, resulting in a marked increase or decrease of the vitamin E plasma level, does not influence formation of posttraumatic vasogenic brain edema. PMID- 9199400 TI - Effects of mild hypothermia on nitric oxide synthesis following contusion trauma in the rat. AB - The exact mechanism of hypothermic cerebroprotection after traumatic brain injury (TBI) is not fully understood. The present study was conducted to investigate the effects of mild hypothermia on trauma-induced synthesis of nitric oxide (NO), which has been implicated in the pathogenesis of ischemic brain damage associated with glutamate neurotoxicity. Cerebral contusion was created in the rat parietal cortex by a weight-drop method, and extracellular concentrations of the NO end products nitrite and nitrate were measured using in vivo brain microdialysis and capillary electrophoresis under normothermic (37 degrees C) and mild hypothermic (32 degrees C) conditions. In normothermic animals, the level of NO end products increased markedly 10 min after contusion, reaching a maximum level at 20 min. In the hypothermic rats, such increases were absent. Although it is unknown whether endothelial NO synthase, neuronal NO synthase, or both caused the elevation of the NO end products seen in the normothermic animals, the present results indicate that inhibition of NO synthesis may play a part in hypothermic cerebroprotection following TBI. PMID- 9199401 TI - Structural relationships in the OmpR family of winged-helix transcription factors. AB - OmpR, a protein that regulates expression of outer membrane porin proteins in enteric bacteria, belongs to a large family of transcription factors. These transcription factors bind DNA and interact productively with RNA polymerase to activate transcription. The two functions, DNA-binding and transcriptional activation, have been localized within the 100 amino acid DNA-binding domain that characterizes members of the OmpR family. Both DNA binding and transcriptional activation by OmpR related proteins have remained poorly understood for lack of structural information or lack of sequence homology with transcription factors of known three-dimensional structure. The recently determined crystal structures of the Escherichia coli OmpR DNA-binding domain (OmpRc) have defined a new subfamily of "winged-helix-turn-helix" DNA-binding proteins. Structural elements of OmpRc can be assigned functional roles by analogy to other winged-helix DNA-binding proteins. A structure based sequence analysis of the OmpR family of transcription factors indicates specific roles for all conserved amino acid residues. Mutagenesis studies performed on several members of this family, OmpR, PhoB, ToxR and VirG, can now be interpreted with respect to the structure. PMID- 9199402 TI - o29 DNA polymerase residue Lys383, invariant at motif B of DNA-dependent polymerases, is involved in dNTP binding. AB - Bacteriophage o29 DNA polymerase shares with other DNA-dependent DNA polymerases several regions of amino acid homology along the primary structure. Among them, motif B, characterized by the consensus +x3Kx(6-7)YG (+ being a positively charged amino acid), appears to be specifically conserved in those polymerases that use DNA but not RNA as template. In particular, the lysine residue of this motif is invariant in all members of DNA-dependent polymerases. In this paper we report a mutational analysis of this invariant residue of motif B with the construction and characterization of two mutant proteins in the corresponding residue (Lys383) of o29 DNA polymerase. Mutant proteins (K383R and K383P) were overexpressed, purified and analyzed under steady-state conditions. In agreement with the modular organization proposed for o29 DNA polymerase, the exonuclease activity was not affected in either mutant protein. Conversely, mutant K383P showed no detectable capacity to incorporate dNTP substrates using either DNA or TP as primer, although its affinity for DNA was not affected. The conservative substitution of Lys383 by arginine (K383R) resulted in a considerable impairment to use dNTPs, in both processive and non-processive DNA synthesis; the Km for dNTPs being 200-fold higher than that of the wild-type enzyme. Mutant K383R recovered the wild-type polymerase/exonuclease ratio when Mn2+ was used instead of Mg2+ as metal activator, indicating a distorted binding of the [dNTP-metal] chelate at the mutant enzyme active site. The positive charge at residue Lys383 was also critical in the catalysis of deoxynucleotidylation of the terminal protein by o29 DNA polymerase. The results obtained suggest a direct role for the lysine residue in motif B in forming an evolutionarily conserved DNA templated dNTP binding pocket. Additionally, K383R mutant protein was also affected in the progression from protein-primed initiation to DNA elongation, a switch between two modes of priming that characterizes o29 DNA replication. PMID- 9199403 TI - RNA hydration: three nanoseconds of multiple molecular dynamics simulations of the solvated tRNA(Asp) anticodon hairpin. AB - The hydration of the tRNA(Asp) anticodon hairpin was investigated through the analysis of six 500 ps multiple molecular dynamics (MMD) trajectories generated by using the particle mesh Ewald method for the treatment of the long-range electrostatic interactions. Although similar in their dynamical characteristics, these six trajectories display different local hydration patterns reflecting the landscape of the "theoretical" conformational space being explored. The statistical view gained through the MMD strategy allowed us to characterize the hydration patterns around important RNA structural motifs such as a G-U base pair, the anticodon U-turn, and two modified bases: pseudouridine and 1 methylguanine. The binding of ammonium counterions to the hairpin has also been investigated. No long-lived hydrogen bond between water and a 2'-hydroxyl has been observed. Water molecules with long-residence times are found bridging adjacent pro-Rp phosphate atoms. The conformation of the pseudouridine is stiffened by a water-mediated base-backbone interaction and the 1-methylguanine is additionally stabilized by long-lived hydration patterns. Such long-lived hydration patterns are essential to ensure the structural integrity of this hairpin motif. Consequently, our simulations confirm the conclusion reached from an analysis of X-ray crystal structures according to which water molecules form an integral part of nucleic acid structure. The fact that the same conclusion is reached from a static and a dynamic point of view suggests that RNA and water together constitute the biologically relevant functional entity. PMID- 9199404 TI - Mutations in the Res subunit of the EcoPI restriction enzyme that affect ATP dependent reactions. AB - The Res subunits of the type III restriction-modification enzymes share a statistically significant amino acid sequence similarity with several RNA and DNA helicases of the so-called DEAD family. It was postulated that in type III restriction enzymes a DNA helicase activity may be required for local unwinding at the cleavage site. The members of this family share seven conserved motifs, all of which are found in the Res subunit of the type III restriction enzymes. To determine the contribution, if any, of these motifs in DNA cleavage by EcoPI, a type III restriction enzyme, we have made changes in motifs I and II. While mutations in motif I (GTGKT) clearly affected ATP hydrolysis and resulted in loss of DNA cleavage activity, mutation in motif II (DEPH) significantly decreased ATP hydrolysis but had no effect on DNA cleavage. The double mutant R.EcoPIK90R-H229K showed no significant ATPase or DNA restriction activity though ATP binding was not affected. These results imply that there are at least two ATPase reaction centres in EcoPI restriction enzyme. Motif I appears to be involved in coupling DNA restriction to ATP hydrolysis. Our results indicate that EcoPI restriction enzyme does not have a strand separation activity. We suggest that these motifs play a role in the ATP-dependent translocation that has been proposed to occur in the type III restriction enzymes. PMID- 9199405 TI - Reconstitution of human topoisomerase I by fragment complementation. AB - Human topoisomerase I (topo I, 91 kDa) is composed of four major domains; the unconserved and highly charged "N-terminal" domain (24 kDa), the conserved "core" domain (54 kDa), a poorly conserved and positively charged "linker" region (5 kDa), and the highly conserved "C-terminal" domain (8 kDa) which contains the active site tyrosine at position 723. Here we demonstrate that human topo I activity can be reconstituted by mixing a 58 kDa recombinant core domain (residues Lys175 to Ala659) with any one of a series of recombinant C-terminal fragments that range in size from 12 kDa (linker and C-terminal domains, residues Leu658 to Phe765) to 6.3 kDa (C-terminal domain residues Gln713 to Phe765). The C terminal fragments bind tightly to the core domain, forming a 1:1 complex that is stable irrespective of ionic strength (0.01 to 1 M). The reconstituted enzymes are active only over a relatively narrow range of salt concentrations (25 to 200 mM KCl) as compared to the intact topo70 enzyme (missing the N-terminal domain). Under physiological conditions (150 mM KCl and 10 mM Mg2+) they are much more distributive in their mode of action than topo70. The reconstituted enzyme binds DNA with an affinity that is approximately 20-fold lower than that of the intact topo70. In addition, the cleavage/religation equilibrium of the reconstituted enzyme appears to be biased towards religation relative to that of the intact enzyme. Despite differences in the cleavage/religation equilibrium and affinity for DNA, the reconstituted and intact enzymes have identical sequence specificities for the cleavage of duplex DNA or suicide cleavage of oligonucleotide substrates. PMID- 9199406 TI - The steric trigger in rhodopsin activation. AB - Rhodopsin is the seven transmembrane helix receptor responsible for dim light vision in vertebrate rod cells. The protein has structural homology with the other G protein-coupled receptors, which suggests that the tertiary structures and activation mechanisms are likely to be similar. However, rhodopsin is unique in several respects. The most striking is the fact that the receptor "ligand", 11 cis retinal, is covalently bound to the protein and is converted from an "antagonist" to an "agonist" upon absorption of light. NMR studies of rhodopsin and its primary photoproduct, bathorhodopsin, have generated structural constraints that enabled docking of the 11-cis and all-trans retinal chromophores into a low-resolution model of the protein proposed by Baldwin. These studies also suggest a mechanism for how retinal isomerization leads to rhodopsin activation. More recently, mutagenesis studies have extended these results by showing how the selectivity of the retinal-binding site can be modified to favor the all-trans over the 11-cis isomer. The structural constraints produced from these studies, when placed in the context of a high-resolution model of the protein, provide a coherent picture of the activation mechanism, which we show involves a direct steric interaction between the retinal chromophore and transmembrane helix 3 in the region of Gly121. PMID- 9199407 TI - Dual conformations of a T cell receptor V alpha homodimer: implications for variability in V alpha V beta domain association. AB - The crystal structure of a mutant T cell receptor (TCR) V alpha domain containing a grafted third complementarity-determining region (CDR3) from a different V alpha was determined at 2.3 A resolution by molecular replacement using the wild type V alpha structure as a search model. Like the wild-type V alpha domain, the mutant crystallized as a homodimer very similar to TCR V alpha V beta and antibody V(L)V(H) heterodimers, with the CDR loops disposed to form part of the antigen-binding site. However, the relative orientation of the two chains in the mutant V alpha homodimer differs from that in the wild-type by a rotation of 14 degrees such that the buried surface area in the dimer interface of the mutant is 140 A2 less than in the wild-type. While the residues forming the interface are essentially the same in the two structures, there are only four pairs of interface hydrogen bonds in the case of the mutant compared with eight for the wild-type. These results suggest that multiple relative orientations of the V alpha and V beta domains of TCRs may be possible, providing a significant contribution to TCR combining site diversity. PMID- 9199408 TI - The second Kunitz domain of human tissue factor pathway inhibitor: cloning, structure determination and interaction with factor Xa. AB - Tissue Factor Pathway Inhibitor (TFPI) is a 36 kDa glycoprotein that helps maintain haemostasis by inhibiting Factor Xa and the Factor VIIa/Tissue Factor (TF) complex. TFPI contains three tandemly linked Kunitz inhibitor domains, of which the second inhibits factor Xa. We have undertaken a multidisciplinary approach to study the structure and function of the second Kunitz domain of TFPI, with a view towards the rational design of factor Xa inhibitors. Amino acid residues 93 to 154 of the mature TFPI protein, corresponding to the second Kunitz domain (TFPI-kII), were expressed in Escherichia coli. The protein was purified to near homogeneity by ion exchange, hydrophobic interaction, and size exclusion chromatography, respectively. TFPI-kII is a potent factor Xa inhibitor with a Ki of 1.5 x 10(-10) M, a value that does not differ significantly from that of intact TFPI. The three-dimensional structure of TFPI-kII in aqueous solution was determined by 1H nuclear magnetic resonance spectroscopy (NMR). A set of 30 conformers was calculated with the program DIANA using 906 distance constraints derived from nuclear Overhauser effects and 23 dihedral angle constraints. This set, representing the solution structure of TFPI-kII, has an average root-mean square deviation of 0.78 A for the backbone atoms and 1.38 A for all heavy atoms of residues 1 to 58. The structure of TFPI-kII has also been determined in complex with porcine trypsin using X-ray crystallographic techniques. The complex has been solved to a resolution of 2.6 A, with a final R-factor of 16.2%. Comparison of the NMR derived structure with that of TFPI-kII in complex with trypsin reveals little divergence of the two structures, with the exception of residue Tyr17. Superposition of the trypsin:TFPI-kII complex on factor Xa provides insights into macromolecular determinants for the inhibition of factor Xa. Complexation would require a degree of reorganisation of factor Xa residues, in particular of TyrF99, but also perhaps of the F148-loop. The interaction was further investigated using restrained molecular dynamics. Electrostatic interactions would appear to play a major role. The reorganisation of factor Xa is in contrast to the proposed factor Xa:TAP interaction, where TAP would bind to the "ground state" structure of factor Xa. PMID- 9199409 TI - Automated NOESY interpretation with ambiguous distance restraints: the refined NMR solution structure of the pleckstrin homology domain from beta-spectrin. AB - We have used a novel, largely automated, calculation method to refine the NMR solution structure of the pleckstrin homology domain of beta-spectrin. The method is called ARIA for Ambiguous Restraints for Iterative Assignment. The starting point for ARIA is an almost complete assignment of the proton chemical shifts, and a list of partially assigned NOEs, mostly sequential and secondary structure NOEs. The restraint list is then augmented by automatically interpreting peak lists generated by automated peak-picking. The central task of ARIA is the assignment of ambiguous NOEs during the structure calculation using a combination of ambiguous distance restraints and an iterative assignment strategy. In addition, ARIA calibrates ambiguous NOEs to derive distance restraints, merges overlapping data sets to remove duplicate information, and uses empirical rules to identify erroneous peaks. While the distance restraints for the structure calculations were exclusively extracted from homonuclear 2D experiments, ARIA is especially suited for the analysis of multidimensional spectra. Applied to the pleckstrin homology domain, ARIA generated structures of good quality, and of sufficiently high accuracy to solve the X-ray crystal structure of the same domain by molecular replacement. The comparison of the free NMR solution structure to the X-ray structure, which is complexed to D-myo-inositol-1,4,5 triphosphate, shows that the ligand primarily induces a disorder-order transition in the binding loops, which are disordered in the NMR ensemble but well ordered in the crystal. The structural core of the protein is unaffected, as evidenced by a backbone root-mean-square difference between the average NMR coordinates and the X-ray crystal structure for the secondary structure elements of less than 0.6 A. PMID- 9199410 TI - Recognition of analogous and homologous protein folds: analysis of sequence and structure conservation. AB - An analysis was performed on 335 pairs of structurally aligned proteins derived from the structural classification of proteins (SCOP http://scop.mrc lmb.cam.ac.uk/scop/) database. These similarities were divided into analogues, defined as proteins with similar three-dimensional structures (same SCOP fold classification) but generally with different functions and little evidence of a common ancestor (different SCOP superfamily classification). Homologues were defined as pairs of similar structures likely to be the result of evolutionary divergence (same superfamily) and were divided into remote, medium and close sub divisions based on the percentage sequence identity. Particular attention was paid to the differences between analogues and remote homologues, since both types of similarities are generally undetectable by sequence comparison and their detection is the aim of fold recognition methods. Distributions of sequence identities and substitution matrices suggest a higher degree of sequence similarity in remote homologues than in analogues. Matrices for remote homologues show similarity to existing mutation matrices, providing some validity for their use in previously described fold recognition methods. In contrast, matrices derived from analogous proteins show little conservation of amino acid properties beyond broad conservation of hydrophobic or polar character. Secondary structure and accessibility were more conserved on average in remote homologues than in analogues, though there was no apparent difference in the root-mean-square deviation between these two types of similarities. Alignments of remote homologues and analogues show a similar number of gaps, openings (one or more sequential gaps) and inserted/deleted secondary structure elements, and both generally contain more gaps/openings/deleted secondary structure elements than medium and close homologues. These results suggest that gap parameters for fold recognition should be more lenient than those used in sequence comparison. Parameters were derived from the analogue and remote homologue datasets for potential used in fold recognition methods. Implications for protein fold recognition and evolution are discussed. PMID- 9199411 TI - Cytochrome cd1 structure: unusual haem environments in a nitrite reductase and analysis of factors contributing to beta-propeller folds. AB - The central tunnel of the eight-bladed beta-propeller domain of cytochrome cd1 (nitrite reductase) is seen, from a 1.28 A resolution structure, to contain hydrogen donors and acceptors that are satisfied by interaction either with water or the d1 haem. The d1 haem, although bound by an extensive network of hydrogen bonds, is not distorted in its binding pocket and is confirmed to have exactly the dioxoisobacteriochlorin structure proposed from chemical studies. A biological rationale is advanced for the undistorted structure of the d1 haem and the large number of hydrogen bonds it makes. The beta-propeller domain can be closely superimposed on that of methanol dehydrogenase despite the enzymes sharing no common sequence motifs and using a different set of interactions to "Velcro" close the propeller. The sequence and likely structural relationships between cytochrome cd1 or methanol dehydrogenase and other predicted eight-bladed beta-propeller domains in proteins, such as the pyrolloquinoline quinone dependent alcohol dehydrogenase, are discussed and compared with other propeller proteins. From sequencing the nirS gene of Thiosphaera pantotropha, it is established that the amino acid sequence deduced previously in part from X-ray diffraction data at lower resolution was largely correct, as was the proposal that eight N-terminal amino acid residues were not seen in the structure. The unusual haem iron environments in both the c-type cytochrome domain, with His/His coordination, and the d1-type cytochrome domain with Tyr/His coordination are related to the functions of the redox centres. PMID- 9199412 TI - Localization of intravenously administered verocytotoxins (Shiga-like toxins) 1 and 2 in rabbits immunized with homologous and heterologous toxoids and toxin subunits. AB - Rabbits challenged intravenously with Shiga toxin or with Escherichia coli verocytotoxin 1 or 2 (VT1 or VT2) are known to develop diarrhea, paralysis, and death, which can be prevented by immunization with a toxoid. The pathological effects of VT1 in the central nervous system and the gastrointestinal tract of unimmunized rabbits correlate with the localization of 125I-VT1 in these tissues, whereas in immunized animals, localization of 125I-VT1 in target tissues is inhibited and labeled toxin is cleared by the liver and spleen. By using the approach described above in this study, rabbits immunized with VT1 toxoid, VT2 toxoid, or with the A or B subunit of each toxin were challenged with intravenous 125I-VT1 or 125I-VT2. After 2 h, the animals were sacrificed, and selected tissues were analyzed for uptake of labeled toxin. It was found that animals immunized with either VT1 toxoid or VT2 toxoid were protected from target tissue uptake of both 125I-VT1 and 125I-VT2. Rabbits immunized with either the VT1 A or VT2 A subunit were also protected from target tissue uptake of both the homologous and heterologous 125I-labeled holotoxins. In contrast, in animals immunized with the toxin B subunits, protection extended only against challenge by the homologous toxin. These results provide evidence of VT1 and VT2 cross neutralization in vivo in the rabbit model and indicate that the in vivo cross neutralization is a function, mainly, of antibodies directed to the VT A subunits. This suggests that the VT1 A or VT2 A subunit may be a suitable immunogen for immunizing humans against systemic VT-mediated disease. PMID- 9199413 TI - Role of neutrophils in experimental septicemia and septic arthritis induced by Staphylococcus aureus. AB - We have previously described a murine model of hematogenously induced Staphylococcus aureus sepsis and arthritis. In this model, large numbers of granulocytes can be observed both in the circulation and locally in the inflamed synovium within 24 h after bacterial inoculation. To assess the role of neutrophils in this severe infection, mice were given granulocyte-depleting monoclonal antibody RB6-8C5 before being inoculated with S. aureus. All the control mice survived their intravenous injection with 3 x 10(7) CFU of S. aureus, whereas all the mice given RB6-8C5 antibody died of sepsis within 2 to 3 days. Even when the inoculum size was reduced sixfold (i.e., 6 x 10(6) CFU/mouse), 50% of the RB6-8C5-treated animals died within 6 days. The RB6-8C5 treated mice had a significantly higher burden of bacteria in their blood and kidneys 24 and 48 h after bacterial inoculation. In addition, when a suboptimal dose of bacteria was administered, the neutrophil-depleted animals displayed a higher frequency of arthritis than did the controls. The granulocyte-depleted animals exhibited increased levels of the proinflammatory cytokines tumor necrosis factor alpha, interleukin-6, and gamma interferon, reflecting the severity of their disease. This is the first direct demonstration of neutrophils playing a crucial protective role in the early phase of S. aureus infection. PMID- 9199414 TI - The tumor necrosis factor alpha-stimulating region of galactose-inhibitable lectin of Entamoeba histolytica activates gamma interferon-primed macrophages for amebicidal activity mediated by nitric oxide. AB - Entamoeba histolytica adheres via galactose-lectin (Gal-lectin) to human colonic mucins and intestinal epithelial cells as a prerequisite to amebic invasion. Native Gal-lectin is a protective antigen in the gerbil model of amebiasis. Amino acids 596 to 1082 of Gal-lectin mediate E. histolytica adherence to target cells and stimulate tumor necrosis factor alpha (TNF-alpha) production by naive murine bone marrow macrophages (BMM). Resistance to amebiasis requires an effective cell mediated immune response against E. histolytica trophozoites mediated by nitric oxide (NO) released from activated macrophages. Herein, we determine whether the TNF-alpha-stimulating region of Gal-lectin can activate gamma interferon (IFN gamma)-primed BMM for NO production and amebicidal activity. Native Gal-lectin (100 to 500 ng/ml) stimulated TNF-alpha and inducible nitric oxide synthase (iNOS) mRNA expression in IFN-gamma-primed BMM as did lipopolysaccharide (100 ng/ml). Primed BMM produced TNF-alpha and NO in response to Gal-lectin in a dose dependent manner. Antilectin monoclonal antibody IG7, which recognizes a domain (amino acids 596 to 818) of the TNF-alpha mRNA-stimulating region of Gal-lectin, specifically inhibited TNF-alpha and iNOS mRNA induction and TNF-alpha and NO production by primed BMM in response to Gal-lectin (100 ng/ml). Simultaneous treatment of BMM with IFN-gamma and Gal-lectin (100 ng/ml) activated the cells to kill E. histolytica trophozoites, whereas IFN-gamma treatment alone had no effect. In the presence of monoclonal antibody 1G7 or aminoguanidine (an iNOS inhibitor), NO production and amebicidal activity were inhibited >80%. These results suggest that the TNF-alpha-stimulating region of native Gal-lectin is a potent stimulus of IFN-gamma-primed BMM for NO production, which is essential for host defense against amebiasis. PMID- 9199415 TI - Intimin-dependent binding of enteropathogenic Escherichia coli to host cells triggers novel signaling events, including tyrosine phosphorylation of phospholipase C-gamma1. AB - Enteropathogenic Escherichia coli (EPEC) interactions with HeLa epithelial cells induced the tyrosine phosphorylation of a host protein of approximately 150 kDa, Hp150. Phosphorylation of this protein band was dependent on the interaction of the EPEC protein intimin with epithelial cell surfaces and was correlated with pedestal formation. Hp150 phosphorylation was specifically inhibited by the addition of cytochalasin D, an inhibitor of actin polymerization, although this appeared to be an indirect effect preventing interaction of intimin with its receptor, tyrosine-phosphorylated Hp90, and thus triggering Hp150 phosphorylation. This suggests the involvement of an actin-based movement of membrane-bound tyrosine-phosphorylated Hp90 to allow its interaction with intimin. Analysis of the tyrosine-phosphorylated Hp150 protein demonstrated that it is heterogeneous in composition, with phospholipase C-gamma1 (PLC-gamma1) being a minor component. Activation of PLC-gamma1 by tyrosine phosphorylation leads to inositol triphosphate and Ca2+ fluxes, events detected following EPEC infection. EPEC also induced tyrosine dephosphorylation of host proteins, including a 240-kDa host protein (Hp240), following EPEC infection. Protein dephosphorylation appears to be a signaling event which occurs independently of intimin. Inhibition of host tyrosine dephosphorylation events by the addition of the tyrosine phosphatase inhibitor sodium vanadate did not prevent actin accumulation beneath the adherent bacteria. We conclude that EPEC induces two sets of signaling events following infection. One set is dependent on EPEC proteins secreted by the type III secretion pathway (EspA and EspB) which induces Hp90 tyrosine phosphorylation and dephosphorylation of host phosphotyrosine proteins. The second set, which is also dependent on the first signaling events, requires intimin interaction with its receptor, tyrosine-phosphorylated Hp90, to trigger Hp150 and PLC-gamma1 tyrosine phosphorylation as well as pedestal formation. Inhibition of pedestal formation by tyrosine kinase inhibitors indicates an important role for tyrosine phosphorylation events during EPEC subversion of host processes. PMID- 9199416 TI - Increased intracellular survival of Mycobacterium smegmatis containing the Mycobacterium leprae thioredoxin-thioredoxin reductase gene. AB - The thioredoxin (Trx) system of Mycobacterium leprae is expressed as a single hybrid protein containing thioredoxin reductase (TR) at its N terminus and Trx at its C terminus. This hybrid Trx system is unique to M. leprae, since in all other organisms studied to date, including other mycobacteria, both TR and Trx are expressed as two separate proteins. Because Trx has been shown to scavenge reactive oxygen species, we have investigated whether the TR-Trx gene product can inhibit oxygen-dependent killing of mycobacteria by human mononuclear phagocytes and as such could contribute to mycobacterial virulence. The gene encoding M. leprae TR-Trx was cloned into the apathogenic, fast-growing bacterium Mycobacterium smegmatis. Recombinant M. smegmatis containing the gene encoding TR Trx was killed to a significantly lesser extent than M. smegmatis containing the identical vector with either no insert or a control M. leprae construct unrelated to TR-Trx. Upon phagocytosis, M. smegmatis was shown to be killed predominantly by oxygen-dependent macrophage-killing mechanisms. Coinfection of M. smegmatis expressing the gene encoding TR-Trx together with Staphylococcus aureus, which is known to be killed via oxygen-dependent microbicidal mechanisms, revealed that the TR-Trx gene product interferes with the intracellular killing of this bacterium. A similar coinfection with Streptococcus pyogenes, known to be killed by oxygen-independent mechanisms, showed that the TR-Trx gene product did not influence the oxygen-independent killing pathway. The data obtained in this study suggest that the Trx system of M. leprae can inhibit oxygen-dependent killing of intracellular bacteria and thus may represent one of the mechanisms by which M. leprae can deal with oxidative stress within human mononuclear phagocytes. PMID- 9199417 TI - Mycoplasma synoviae has two distinct phase-variable major membrane antigens, one of which is a putative hemagglutinin. AB - Mycoplasma synoviae is a major pathogen of poultry, causing synovitis and respiratory infection. A cluster of 45- to 50-kDa membrane proteins is immunodominant in strain WVU-1853. Four distinct proteins were identified in this cluster by high-pressure liquid chromatography. Monoclonal antibodies and monospecific antisera against each established that they fell into two groups, MSPA and MSPB, each containing two members distinguishable by a difference in hydrophobicity. A 25- to 30-kDa membrane protein (MSPC) was shown to be antigenically related to the MSPB proteins. Considerable variation in the size and expression of MSPA and MSPB was observed among different strains of M. synoviae. Examination of expression in colonies of strain WVU-1853 established that both MSPA and MSPB (and MSPC) were phase variable. Immunostaining of MSPB (and MSPC) with monoclonal antibodies exhibited quantal variation, with three distinct levels observed between and within colonies. Hemadsorption by M. synoviae colonies was also found to be phase variable, with some colonies exhibiting sectorial expression of hemadsorption. Monospecific antisera against MSPA inhibited hemagglutination, but neither monoclonal antibodies nor monospecific antisera against MSPB could inhibit hemagglutination. However, loss of the capacity to hemadsorb by individual clones was associated with loss of expression of both MSPA and MSPB. These findings have elucidated the complexity of structure, function, and expression of the 45- to 50-kDa membrane protein cluster of M. synoviae, and they suggest that all members of the cluster may be involved in adhesion. PMID- 9199418 TI - Passive transfer of a monoclonal antibody specific for a sialic acid-dependent epitope on the surface of Trypanosoma cruzi trypomastigotes reduces infection in mice. AB - Trypanosoma cruzi, the parasite that causes Chagas' disease, proliferates in the cytosol of mammalian cells. When the trypomastigote forms exit the infected cell, they become extensively sialylated because the parasite contains an enzyme called trans-sialidase. This enzyme efficiently catalyzes the transfer of bound sialic acid residues from host glycoconjugates to acceptors containing terminal beta galactosyl residues on the parasite surface. The sialic acid acceptors are developmentally regulated mucin-like glycoproteins that are extremely abundant on the trypomastigote surface. In the present study, we determined whether passive transfer of monoclonal antibodies specific for sialic acid acceptors could reduce the acute infection induced by T. cruzi in a highly susceptible mouse strain. We found that passive transfer to naive mice of an immunoglobulin G1 monoclonal antibody directed to a sialylated epitope of these mucin-like glycoproteins significantly decreased parasitemia and the number of tissue parasites as measured by a DNA probe specific for T. cruzi. Upon challenge with trypomastigotes, mice which received this antibody also had a significant increase in survival. A statistically significant reduction in parasitemia could be accomplished with relatively small doses of immunoglobulin, and Fab fragments alone could not mediate protective immunity. The precise mechanism of parasite elimination is unknown; however, this monoclonal antibody does not lyse trypomastigotes in vitro in the presence of human complement or mouse spleen cells. PMID- 9199419 TI - Enteropathogenic Escherichia coli O103 from rabbit elicits actin stress fibers and focal adhesions in HeLa epithelial cells, cytopathic effects that are linked to an analog of the locus of enterocyte effacement. AB - Escherichia coli O103, a major agent of weaned-rabbit diarrhea in Western Europe, was previously shown to produce diarrhea and attaching-and-effacing intestinal lesions in experimentally infected rabbits and to possess a homolog of the eaeA gene of enteropathogenic E. coli (EPEC). In the present study, we have shown that although negative in the fluorescent-actin staining test on HeLa cells, prototype rabbit E. coli O103 strain B10 was able to induce an original cytopathic effect (CPE) in the same interaction model. This CPE was characterized by a generalized reorganization of the actin cytoskeleton and the formation of focal adhesions on the entire surface of the target cells. These effects amplified with time, leading to cell death about 5 days after the interaction. They were produced by all rabbit E. coli O103 strains tested, by rabbit-infecting E. coli RDEC-1, and also by two human EPEC isolates. We localized genes associated with CPE by using TnphoA insertion mutagenesis in strain B10. In all five independent CPE-negative mutants that we were able to generate, the insertion was located in a region of the genome homologous to the 35-kb locus of enterocyte effacement (LEE locus) of EPEC E2348/69. The mutants concurrently lost the ability to secrete four major supernatant proteins of 25, 37, 39, and 40 kDa, which were shown by immunoprecipitation to share antigenic determinants with secreted proteins of human EPEC E2348/69. The virulence of one of these mutants (strain B10/CA1) was compared with that of the parental strain in the weaned-rabbit diarrhea model. The mutant was totally deprived of virulence, although it colonized the intestine as efficiently as the parental strain did. This study points to a new pathogenic trait of EPEC strains, which is associated with the LEE locus and, possibly, with in vivo virulence. PMID- 9199420 TI - Intrapulmonary response to Histoplasma capsulatum in gamma interferon knockout mice. AB - We examined the immunobiological responses to Histoplasma capsulatum in lungs of gamma interferon (IFN-gamma) knockout mice (GKO mice). Naive GKO mice succumbed by day 9 to intranasal challenge with 2.5 x 10(6) yeasts, whereas all wild-type (WT) mice survived for 45 days. Compared to lungs of WT mice, the lungs of acutely infected GKO mice exhibited dramatically elevated numbers of CFU in lungs and significantly higher levels of tumor necrosis factor alpha (TNF-alpha) and granulocyte-macrophage colony-stimulating factor (GM-CSF) but not interleukin-12 (IL-12) or IL-4. To determine if IFN-gamma is necessary in reexposure histoplasmosis, GKO and WT mice were inoculated with 10(4) yeasts intranasally and given amphotericin B for 3 weeks. Six weeks later, mice were rechallenged with 2.5 x 10(6) yeasts. All GKO mice died by day 6, whereas all WT mice survived for 45 days. Lungs of GKO mice contained substantially elevated numbers of CFU and higher TNF-alpha and GM-CSF levels but not IL-12 or IL-4. Thus, IFN-gamma is requisite for control of pulmonary histoplasmosis in naive and reexposed mice. PMID- 9199421 TI - Internalization of Escherichia coli by human renal epithelial cells is associated with tyrosine phosphorylation of specific host cell proteins. AB - Human renal epithelial cells are capable of internalizing Escherichia coli regardless of whether the bacteria are isolated from individuals with pyelonephritis or from healthy volunteers. In this study, we investigated the role of host cell tyrosine kinase activity in internalization. We found that internalization of both fecal and pyelonephritis isolates is blocked by tyrosine kinase inhibitors. We found increased intensity of two tyrosine-phosphorylated proteins, with relative mobilities of approximately 123,000 and 110,000, in Western blots of extracts from human renal epithelial cells infected with E. coli. The increased intensity of these tyrosine-phosphorylated proteins was observed only in the Triton X-100-insoluble fraction, suggesting that these proteins could be associated with the cytoskeleton. Increased tyrosine phosphorylation of these proteins upon E. coli infection was observed in both transformed and primary human renal epithelial cells and in cells infected with several different strains of E. coli isolated from the feces of healthy individuals or from the blood or urine of patients with pyelonephritis. The increased tyrosine phosphorylation of these proteins required live bacteria and was blocked by tyrosine kinase inhibition but not by protein synthesis inhibitors or cytochalasin D. These experiments establish a strong link between E. coli internalization and host cell signaling through tyrosine kinases in human kidney cells and provide evidence that specific proteins are involved in these processes. PMID- 9199422 TI - Regulation of anthrax toxin activator gene (atxA) expression in Bacillus anthracis: temperature, not CO2/bicarbonate, affects AtxA synthesis. AB - Anthrax toxin gene expression in Bacillus anthracis is dependent on the presence of atxA, a trans-acting regulatory gene located on the resident 185-kb plasmid pXO1. In atxA+ strains, expression of the toxin genes (pag, lef, and cya) is enhanced by two physiologically significant signals: elevated CO2/bicarbonate and temperature. To determine whether increased toxin gene expression in response to these signals is associated with increased atxA expression, we monitored steady state levels of atxA mRNA and AtxA protein in cells cultured in different conditions. We purified histidine-tagged AtxA [AtxA(His)] from Escherichia coli and used anti-AtxA(His) serum to detect AtxA in protein preparations from B. anthracis cells. AtxA was identified as a protein with an apparent size of 56 kDa in cytoplasmic fractions of B. anthracis cells. Our data indicate that atxA expression is not influenced by CO2/bicarbonate levels. However, the steady-state level of atxA mRNA in cells grown in elevated CO2/bicarbonate at 37 degrees C is five- to sixfold higher than that observed in cells grown in the same conditions at 28 degrees C. A corresponding difference in AtxA protein was also seen at the different growth temperatures. When atxA was cloned on a multicopy plasmid in B. anthracis, AtxA levels corresponding to the atxA gene copy number were observed. However, this strain produced significantly less pag mRNA and protective antigen protein than the parental strain harboring atxA in single copy on pXO1. These results indicate that increased AtxA expression does not lead to a corresponding increase in pag expression. Our data strongly suggest that an additional factor(s) is involved in regulation of pag and that the relative amounts of such a factor(s) and AtxA are important for optimal toxin gene expression. PMID- 9199423 TI - Unimpaired down-modulation of the hepatic granulomatous response in CD8 T-cell- and gamma interferon-deficient mice chronically infected with Schistosoma mansoni. AB - The granulomatous response to schistosome eggs is a CD4 T-cell-dependent, Th2 cytokine-dominated immunopathologic response. As infection proceeds to chronicity, both granuloma formation and egg-induced cytokine production become downregulated, and previous experiments have implicated CD8 T cells in this process. One mechanism by which CD8 T cells could suppress immunopathology is through the production of the counterregulatory cytokine gamma interferon (IFN gamma), but no in vivo evidence exists to directly support this hypothesis. In this study, we analyzed hepatic granuloma formation and egg-induced cytokine production in Schistosoma mansoni-infected gene knockout mice deficient in either CD8 lymphocytes or IFN-gamma. Surprisingly, we found that neither immunologic component plays an essential function in the control of granuloma and cytokine responses during either the acute or chronic stage of infection. Thus, other mechanisms may be more important in the regulation of immunopathology in schistosomiasis. PMID- 9199424 TI - Predominant recognition of the ESAT-6 protein in the first phase of interferon with Mycobacterium bovis in cattle. AB - Tuberculosis continues to be a worldwide health problem for both humans and animals. The development of improved vaccines and diagnostic tests requires detailed understanding of the immune responses generated and the antigens recognized during the disease. This study examined the T-cell response which develops in cattle experimentally infected with Mycobacterium bovis. The first significant T-cell response was found 3 weeks after the onset of infection and was characterized by a pronounced gamma interferon (IFN-gamma) response from peripheral blood mononuclear cells directed to antigens in culture filtrates. Short-term culture filtrate (ST-CF) was separated into molecular mass fractions and screened for recognition by T cells from experimentally infected and field cases of bovine tuberculosis. Cattle in the early stages of experimental infection were characterized by strong IFN-gamma responses directed predominantly toward the lowest-mass (<10-kDa) fraction of ST-CF, but cattle in later stages of experimental infection (16 weeks postinfection) exhibited a broader recognition of antigens of various molecular masses. Field cases of bovine tuberculosis, in comparison, preferentially recognized low-mass antigens, characteristic of animals in the early stages of infection. The major T-cell target for this dominant IFN-gamma response was found to be the secreted antigen ESAT-6. This antigen was recognized strongly by the majority of field cases of bovine tuberculosis tested. As ESAT-6 is unique to pathogenic mycobacterial species, our study suggests that ESAT-6 is an antigen with major potential for vaccination against and specific diagnosis of bovine tuberculosis. PMID- 9199425 TI - Generation of targeted nonpolar gene insertions and operon fusions in Pasteurella haemolytica and creation of a strain that produces and secretes inactive leukotoxin. AB - An efficient method for targeted gene inactivation and generation of chromosomal gene fusions in Pasteurella haemolytica has been devised and used to create an lktC::cat operon fusion by allelic exchange at the leukotoxin gene cluster (lktCABD). A copy of the lktC gene was insertionally inactivated by using a nonpolar, promoterless cat cassette and then delivered into P. haemolytica on a shuttle vector. Plasmid incompatibility was used to detect clones where double recombination events had occurred at the chromosomal locus. The insertion in lktC did not affect expression of the downstream genes, and the mutant strain secreted an antigenic proleukotoxin that was neither leukotoxic nor hemolytic. Expression of the lktC gene in trans restored the wild-type phenotype, confirming that LktC is required for activation of the proleukotoxin to the mature leukotoxin. Construction of the lktC::cat operon fusion allowed us to quantitate leukotoxin promoter activity in P. haemolytica and to demonstrate that transcription was maximal during early logarithmic growth phase but was reduced following entry into late logarithmic phase. This allelic exchange system should be useful for future genetic studies in P. haemolytica and could potentially be applied to other members of Haemophilus-Actinobacillus-Pasteurella family, where genetic manipulation is limited. PMID- 9199426 TI - Extracellular proteins of Cryptococcus neoformans and host antibody response. AB - Proteins secreted by the fungal pathogen Cryptococcus neoformans may be involved in invasion and could be useful in vaccine design. Despite the medical importance of this fungus, little is known about its extracellular proteins or the immune response to these antigens. To study C. neoformans extracellular proteins, 12 strains were metabolically radiolabeled and protein supernatants were analyzed. Both strain- and growth condition-dependent differences were observed. Enzymatic assays of filtered culture supernatants revealed butyrate esterase and caprylate esterase lipase activity for 11 of 12 strains, as well as acid phosphatase, naphthol-AS-BI-phosphohydrolase, and beta-glucosidase activities in some strains. Serum from infected rodents immunoprecipitated several secreted proteins, consistent with in vivo expression and development of an antibody response. For strain 24067, two immunodominant species, of approximately 75 and 30 kDa, were recognized. The relative intensity of the autoradiographic bands depended on the route of infection for both rats and mice. In summary, our results indicate that (i) there are multiple proteins in C. neoformans culture supernatants, (ii) there are strain differences in supernatant protein profiles, (iii) there are differences in supernatant protein profile depending on the growth conditions, (iv) there are several new extracellular and/or cell-associated enzymatic activities, and (v) antibodies to several supernatant proteins are made in the course of infection. PMID- 9199427 TI - Enteropathogenic Escherichia coli protein secretion is induced in response to conditions similar to those in the gastrointestinal tract. AB - The pathogenicity of enteropathogenic Escherichia coli (EPEC) is associated with the expression and secretion of specific bacterial factors. EspB is one such secreted protein which is required to trigger host signaling pathways resulting in effacement of microvilli and cytoskeletal rearrangements. These events presumably contribute to the ensuing diarrhea associated with EPEC infections. EPEC encounters several environmental changes and stimuli during its passage from the external environment into the host gastrointestinal tract. In this paper we show that the secretion of EspB is subject to environmental regulation, and maximal secretion occurs under conditions reminiscent of those in the gastrointestinal tract. Thus, secretion is maximal at 37 degrees C, pH 7, and physiological osmolarity. In addition, maximal secretion requires the presence of sodium bicarbonate and calcium and is stimulated by millimolar concentrations of Fe(NO3)3. The secretion of the four other EPEC-secreted proteins appears to be modulated in a manner similar to that of EspB. Our results also show that secretion is not dependent on CO2, as originally reported by Haigh et al. (FEMS Microbiol. Lett. 129: 63-67, 1995), but that CO2 more likely acts as a component of the medium buffering system, since CO2 dependence was abolished by the use of alternative buffers. PMID- 9199428 TI - Characterization of a class II pilin expression locus from Neisseria meningitidis: evidence for increased diversity among pilin genes in pathogenic Neisseria species. AB - Strains of Neisseria meningitidis elaborate one of two classes of pili. Meningococcal class I pili have many features in common with pili produced by N. gonorrhoeae, including the ability to bind monoclonal antibody SM1 and a common gene and protein structure consisting of conserved, semivariable, and hypervariable regions. Class II pili are SM1 nonreactive and display smaller subunit molecular weights than do gonococcal or meningococcal class I pili. In this study, we have determined the N-terminal amino acid sequence for class II pilin and isolated the expression locus encoding class II pilin from N. meningitidis FAM18. Meningococcal class II pilin displays features typical of type IV pili and shares extensive amino acid identity with the N-terminal conserved regions of other neisserial pilin proteins. However, the deduced class II pilin sequence displays several unique features compared with previously reported meningococcal class I and gonococcal pilin sequences. Class II pilin lacks several conserved peptide regions found within the semivariable and hypervariable regions of other neisserial pilins and displays a large deletion in a hypervariable region of the protein believed to be exposed on the pilus face in gonococcal pili. DNA sequence comparisons within all three regions of the coding sequence also suggest that the meningococcal class II pilin gene is the most dissimilar of the three types of neisserial pilE loci. Additionally, the class II locus fails to display flanking-sequence homology to class I and gonococcal genes and lacks a downstream Sma/Cla repeat sequence, a feature present in all other neisserial pilin genes examined to date. These data indicate meningococcal class II pili represent a structurally distinct class of pili and suggest that relationships among pilin genes in pathogenic Neisseria do not necessarily follow species boundaries. PMID- 9199429 TI - Modification of the Staphylococcus aureus fibronectin binding phenotype by V8 protease. AB - The amount of cell surface fibronectin (Fn)-binding protein (FnBP) adhesin expressed by Staphylococcus aureus is maximal during exponential growth but disappears rapidly as the culture progresses into stationary phase. To identify factors responsible for the loss of cell surface FnBP, a culture of S. aureus L170, which shows high levels of Fn binding, was supplemented at the time of inoculation with concentrated stationary-phase supernatant from S. aureus L530, a strain which binds Fn poorly. The resulting exponential-phase cells were devoid of FnBP. The factor responsible for this activity was purified from the culture supernatant and identified as V8 protease. When cultured with 375 ng of exogenous V8 protease ml(-1), exponential-phase cells of S. aureus L170 were devoid of cell surface FnBP, and concentrations as low as 23 ng x ml(-1) resulted in reduced amounts of FnBP. Addition of the protease inhibitor alpha2-macroglobulin to the culture medium prevented the growth-phase-dependent loss of cell surface FnBP, whereas growth with exogenous V8 protease resulted in reduced adherence to the solid-phase N-terminal fragment of Fn and to the extracellular matrix synthesized by fetal rabbit lung fibroblasts. Although FnBP was extremely sensitive to V8 protease, exogenous protease did not exert a significant influence on the amount of cell surface protein A. However, a limited number of other high-molecular weight cell surface proteins were also sensitive to V8 protease. Therefore, both the adhesive phenotype and cell surface protein profile of S. aureus can be modified by V8 protease activity. PMID- 9199430 TI - Synthesis and function of Actinomyces naeslundii T14V type 1 fimbriae require the expression of additional fimbria-associated genes. AB - The nucleotide sequence of the chromosomal DNA flanking the Actinomyces naeslundii (formerly A. viscosus) T14V type 1 fimbrial structural subunit gene (fimP) was determined. Six open reading frames (ORFs), in the order 5' ORF3, ORF2, ORF1,fimP, ORF4, ORF5, ORF6 3', were identified. ORF1 encoded a protein of 408 amino acid residues (Mr = 39,270) and had significant sequence homology with the A. naeslundii T14V type 1 and A. naeslundii WVU45 type 2 fimbrial structural subunits. An in-frame fusion of ORF1 to the malE gene of the expression vector, pMAL-c2, yielded a protein that was immunostained with antibodies raised against the maltose binding protein and A. naeslundii T14V whole bacteria. Digestion of the fusion protein with factor Xa released a protein (apparent molecular mass of 34 kDa) that was immunostained only with the antibody directed against A. naeslundii T14V whole bacterial cells. Integration plasmids carrying a kanamycin resistance gene (kan) that was used to substitute for ORF1 or for DNA fragments internal to the coding region of the other five ORFs were used to transform A. naeslundii T14V. Neither type 1 fimbriae nor the 65-kDa fimbrial structural subunit was detected in mutants obtained by allelic replacement of ORF1 or ORF2. Mutants obtained by allelic replacement of ORF3 or ORF4 expressed only the 65-kDa fimbrial structural subunit. These mutants did not bind, in vitro, to proline rich proteins that serve as the receptors for Actinomyces type 1 fimbriae. In contrast, a mutant in which the integration plasmid DNA had been inserted at a site close to the carboxyl terminus of ORF6 expressed type 1 fimbriae and had adherence properties similar to those observed in the wild-type strain. These results demonstrate the existence of additional genes near fimP that are likely to be involved in the synthesis and function of cell surface fimbriae of A. naeslundii T14V. PMID- 9199431 TI - Analysis of F1F0-ATPase from Helicobacter pylori. AB - The adaptive mechanisms that permit Helicobacter species to survive within the gastric mucosa are not well understood. The proton-translocating F1F0-ATPase is an important enzyme for regulating intracellular pH or synthesizing ATP in many other enteric bacteria; therefore, we used degenerate primers derived from conserved bacterial F1F0-ATPase sequences to PCR amplify and clone the gene (atpD) encoding the H. pylori F1F0-ATPase beta subunit. The deduced amino acid sequences of the F1F0-ATPase beta subunits from H. pylori and Wolinella succinogenes were 85% identical (91% similar). To characterize a potential functional role of F1F0-ATPase in H. pylori, H. pylori or Escherichia coli cells were incubated for 60 min in buffered solutions at pH 7, 6, 5, or 4, with or without 100 microM N,N'-dicyclohexylcarbodiimide (DCCD), a specific inhibitor of F1F0-ATPase. At pH 5 and 4, there was no significant decrease in survival of H. pylori in the presence of DCCD compared to its absence, whereas incubation with DCCD at pH 7 and 6 significantly decreased H. pylori survival. E. coli survival was unaffected by DCCD at any pH value tested. We next disrupted the cloned beta subunit sequence in E. coli by insertion of a kanamycin resistance cassette and sought to construct an isogenic F1F0-ATPase H. pylori mutant by natural transformation and allelic exchange. In multiple transformations of H. pylori cells grown at pH 6 or 7, no kanamycin-resistant F1F0 mutants were isolated, despite consistently successful mutagenesis of other H. pylori genes by using a similar approach and PCR experiments providing evidence for integration of the kanamycin resistance cassette into atpD. The sensitivity of H. pylori to DCCD at pH 7 and 6, and failure to recover F1F0 H. pylori mutants under similar conditions, suggests that the function of this enzyme is required for survival of H. pylori at these pHs. PMID- 9199432 TI - Nitrite reductase from Pseudomonas aeruginosa induces inflammatory cytokines in cultured respiratory cells. AB - Persistent infection with Pseudomonas aeruginosa increases interleukin-8 (IL-8) levels and causes dense neutrophil infiltrations in the airway of patients with chronic airway diseases. To investigate the role of P. aeruginosa infection in IL 8 production in the airway of these patients, we examined whether cell lysates of P. aeruginosa could cause IL-8 production from human bronchial epithelial cells. Diluted sonicated supernatants of P. aeruginosa (SSPA) with a mucoid or nonmucoid phenotype stimulated human bronchial epithelial (BET-1A) cells to produce IL-8. In this study, we have purified a 59-kDa heat-stable protein with IL-8-inducing activity from the SSPA by sequential ion-exchange chromatography. The N-terminal sequence of this purified protein completely matched a sequence at the N-terminal part of the mature protein of nitrite reductase from P. aeruginosa. In addition, immunoblotting with a polyclonal immunoglobulin G (IgG) against recombinant Pseudomonas nitrite reductase demonstrated a specific binding to the purified protein. Furthermore, the immunoprecipitates of the SSPA with a polyclonal IgG against recombinant nitrite reductase induced a twofold-higher IL-8 production in the BET-1A cell culture than did the immunoprecipitates of the SSPA with a control IgG. These lines of evidence confirmed that Pseudomonas nitrite reductase was responsible for IL-8 production in the BET-1A cells. The purified nitrite reductase induced maximal expression of IL-8 mRNA in the BET-1A cells at 1 to 3 h after stimulation, and the IL-8 mRNA expression declined by 8 h after stimulation. New protein translation was not required for nitrite reductase mediated IL-8 mRNA expression in the BET-1A cells. Nitrite reductase stimulated the BET-1A cells, as well as human alveolar macrophages, pulmonary fibroblasts, and neutrophils, to produce IL-8. In contrast, nitrite reductase induced significant levels of tumor necrosis factor alpha and IL-1beta protein only in human alveolar macrophages. These data support the notion that nitrite reductase from P. aeruginosa induces the production of inflammatory cytokines by respiratory cells and may contribute to the pathogenesis of chronic airway diseases and persistent P. aeruginosa infection. PMID- 9199433 TI - Staphylococcal enterotoxin A-induced fever is associated with increased circulating levels of cytokines in rabbits. AB - Rabbits were injected intravenously with 10 to 100 ng of staphylococcal enterotoxin A (SEA) per kg, and colonic temperatures were monitored. The febrile responses were compared with circulating levels of interferon (IFN), tumor necrosis factor (TNF), interleukin-1 (IL-1), IL-2, and IL-6 just before the injection of SEA. Both colonic temperatures and circulating levels of IFN, TNF, and IL-2 started to rise at 1 to 2 h and reached their peak levels at 3 to 5 h after SEA injection. Both the fever and the increased circulating levels of IFN, TNF, and IL-2 produced by SEA were decreased by pretreatment with indomethacin (a cyclo-oxygenase inhibitor) (15 mg/kg, intraperitoneally), anisomycin (a protein synthesis inhibitor) (15 mg/kg, subcutaneously), or dexamethasone (an effective anti-inflammatory and immunosuppressive agent) (4 mg/kg, intravenously) in rabbits. Rabbits were injected intravenously with 30 ng of SEA per kg on four consecutive days, and colonic temperatures were monitored. Compared to rabbits that received the single injection of SEA, rabbits that received four consecutive injections of SEA showed a lesser increase in circulating levels of IFN, TNF, and IL-2 as well as colonic temperatures in response to an intravenous dose of SEA (30 ng/kg). The data suggest that the prevention of the febrile response elicited by SEA by indomethacin, anisomycin, or dexamethasone is due to prevention by these compounds of the increase in the circulating levels of IFN, TNF, and IL-2. The pyrogenic hyporesponsiveness to repeated injection of SEA is associated with decreased production of these circulating cytokines. PMID- 9199434 TI - Hyperlipoproteinemia enhances susceptibility to acute disseminated Candida albicans infection in low-density-lipoprotein-receptor-deficient mice. AB - Recent studies have suggested the use of lipoproteins as an adjuvant treatment of lethal gram-negative infections. However, other important microorganisms for the etiology of sepsis, such as Candida species, grow better in lipid-rich environments. We investigated the effect of hyperlipoproteinemia on systemic candidiasis in low-density-lipoprotein-receptor-deficient (LDLR-/-) mice, in which the loss of the receptor results in a seven- to ninefold-higher plasma LDL level than that in their wild-type littermates (C57BL/6J). LDLR-/- mice died earlier, and the outgrowth of Candida albicans in the kidneys and livers of LDLR /- mice was significantly higher compared with that of controls. After infection, circulating cytokine concentrations were significantly higher in LDLR-/- mice. In vitro, C. albicans grew better in plasma samples of LDLR-/- mice than in control plasma samples and peritoneal macrophages of LDLR-/- mice challenged with heat killed C. albicans produced more cytokines than did those of controls. This latter phenomenon was probably due to increased binding of yeast cells to macrophages of LDLR-/- mice. These data suggest that hyperlipoproteinemia is deleterious in systemic candidiasis. PMID- 9199435 TI - Antigenic characterization and analysis of the human immune response to outer membrane protein E of Branhamella catarrhalis. AB - Outer membrane protein E (OMP E) is a 50-kDa major OMP of Branhamella catarrhalis. Polyclonal antisera and four monoclonal antibodies (MAbs) to OMP E were generated to study its antigenic structure. All antibodies recognized epitopes in all 19 B. catarrhalis strains tested by immunoblot assays. By flow cytometry, it was determined that MAbs 1B3 and 9G10d recognized epitopes which are expressed on the surface of the intact bacterium, while MAbs IC11 and 7C10 recognized epitopes which were buried within the outer membrane. A competitive enzyme-linked immunosorbent assay showed that MAbs 1B3 and 9G10d recognize the same or closely related epitopes. Proteinase K treatment of whole bacterial cells revealed that MAbs 1B3 and 9G10d recognize a surface-exposed epitope located in the 17-kDa region towards the amino terminus of OMP E. The human serum and mucosal antibody responses to OMP E in adults with chronic bronchitis were studied. A majority of these patients had immunoglobulin A to OMP E in sputum supernatants. None of ten adults who experienced lower respiratory tract infections due to B. catarrhalis demonstrated a clear-cut rise in antibody titer to OMP E in serum or sputum supernatant. This study has demonstrated that OMP E has at least one surface-exposed epitope which is highly conserved among strains of B. catarrhalis and which is located in the amino-terminal 184 amino acids of the molecule. PMID- 9199436 TI - Intranasal vaccination of humans with recombinant cholera toxin B subunit induces systemic and local antibody responses in the upper respiratory tract and the vagina. AB - Forty-five volunteers were vaccinated twice intranasally with 10, 100, or 1,000 microg of cholera toxin B subunit (CTB). Blood and nasal and vaginal secretions were collected before and 1 week after the second vaccination from all volunteers, and the specific and total immunoglobulin A (IgA) and IgG titers were determined by enzyme-linked immunosorbent assay. Samples were also taken 6 months (n = 16) and 1 year (n = 14) after the vaccination. The 10- and 100-microg doses were well tolerated by the volunteers, but the 1,000-microg dose induced increased secretions from the nose and repetitive sneezings for several hours. The CTB-specific serum IgA and IgG increased 21- and 7-fold, respectively, 1 week after vaccination with the medium dose and increased 61- and 37-fold, respectively, after the high dose. In nasal secretions the specific IgA and IgG increased 2- and 6-fold after the medium dose and 2- and 20-fold after the high dose, respectively. In vaginal secretions the specific IgA and IgG increased 3- and 5-fold after the medium dose and 56- and 74-fold after the high dose, respectively. The lowest dose did not induce any significant antibody titer increases in serum or in secretions. The specific IgA and IgG levels in secretions were still elevated after 6 months but were decreasing 1 year after the vaccination. These results show that intranasal vaccination of humans with CTB induces strong systemic and mucosal antibody responses and suggest that CTB may be used as a carrier for antigens that induce protective immunity against systemic as well as respiratory and genital infections. PMID- 9199437 TI - Evolutionary relationships among pathogenic and nonpathogenic Escherichia coli strains inferred from multilocus enzyme electrophoresis and mdh sequence studies. AB - Within the species Escherichia coli, there are commensal strains and a variety of pathogenic strains, including enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), and urinary tract infection (UTI) strains. The pathogenic strains are identified by serotype and by possession of specific virulence determinants (toxins and adhesions, etc.) encoded by either monocistronic genes, plasmids, or pathogenicity islands. Although there are studies on the relationships between selected pathogenic strains, the relatedness among the majority of the pathogenic forms to each other, to commensal E. coli, and to the genus Shigella (which has often been suggested to be part of E. coli) has not been determined. We used multilocus enzyme electrophoresis (MLEE) at 10 enzyme loci and the sequence of the mdh housekeeping gene to study the genetic relationships of pathogenic E. coli strains (including Shigella clones), namely, 5 EPEC strains (serotypes O111 and O55), 3 EHEC strains (serotype O157), 6 ETEC strains (serotypes O78, O159, and O148), 5 EIEC strains (serotypes O124, O28, and O112), and 13 Shigella strains representing clones Flexneri, Dysenteriae, Boydii, and Sonnei, to commensal E. coli strains. Both the MLEE and mdh sequence trees reveal that EPEC, EHEC, ETEC, EIEC, and UTI strains are distributed among the ECOR set groups, with no overall clustering of EPEC, ETEC, EIEC, or UTI strains. The genus Shigella is shown to comprise a group of closely related pathogenic E. coli strains. Six pathogenic strains, i.e., M502 (EIEC; O112ac:NM), M503 (EPEC; O111:H12), M526 (ETEC; O159:H4), M522 (EPEC; O111ac:H12), M524 (ETEC; O78:H11), and M506 (ETEC; O78:H11), were found to have mdh sequences identical to those of five ECOR group A strains (ECOR5, ECOR10, ECOR14, ECOR6, and K-12). All 11 strains are closely related by MLEE. The results indicate that pathogenic strains of E. coli do not have a single evolutionary origin within E. coli but have arisen many times. The results also suggest the possibility that any E. coli strain acquiring the appropriate virulence factors may give rise to a pathogenic form. PMID- 9199438 TI - Effect of immunoglobulin G isotype on the infectivity of Chlamydia trachomatis in a mouse model of intravaginal infection. AB - It has been previously shown with an in vitro neutralization system that monoclonal antibodies (MAbs) to the major outer membrane protein (MOMP) of Chlamydia trachomatis, depending on the isotype of the MAb and the host cell used, can either neutralize or enhance the infectivity of this organism. MAbs to variable domain 4 (VD 4) of MOMP have been described that neutralize the infectivity of C. trachomatis when tested in a system in which either the host cell does not have detectable Fc gammaRIII receptors or complement is added to block the interaction of the MAb with the receptor. However, if Fc gammaRIII receptors are available, immunoglobulin G2b (IgG2b) MAbs to the VD 4 are able to enhance the infectivity of this pathogen. Two MAbs that recognize the sequence TLNPTIA in VD 4 of the MOMP but differ in isotype, E4 (IgG2b) and E21 (IgG1), were used to test whether in vivo the isotype of the MAb modulates the outcome of a vaginal infection in a murine model. A third MAb, CP33 (IgG2b), that recognizes the chlamydial lipopolysaccharide but does not neutralize infectivity of C. trachomatis, was also tested. Elementary bodies (EBs) of C. trachomatis, serovar E (BOUR), were pretreated with the three MAbs and were used to inoculate the vaginas of C3H/HeJ mice which had been pretreated with progesterone. Subsequently mice were monitored over a 5-week period with vaginal cultures. In the groups that were inoculated with EBs pretreated with MAbs directed to VD 4 of MOMP, there was a significant decrease (P < 0.05) in the number of mice infected. Only 30% of the mice were infected in the MAb E4-treated group, and 10% were infected in the MAb E21 group. This was in contrast to the groups inoculated with EBs pretreated with MAb CP33 and control untreated EBs, which resulted in 100 and 79% of the mice infected, respectively. Therefore, in this setting in which EBs were introduced in vivo coated with MAb, there was no enhancement of infection by IgG2b MAbs; rather, the results paralled the in vitro neutralization results, in which cells lacking Fc gammaRIII receptors were employed. Mice were also given the MAbs, as well as purified IgG as a control, by intraperitoneal injection before and after intravaginal inoculation with C. trachomatis. Despite relatively high levels of MAbs in serum and detectable levels of MAbs in the vagina at the time of infection, there was only modest protection in animals receiving MAb E21, with 60% of the mice infected in contrast to 90% of the mice receiving MAb E4, MAb CP33, and IgG. However, by the second week of infection compared to controls, there was a significant increase (P < 0.05) in the amount of chlamydiae recovered from the vaginas of mice that had received the two IgG2b MAbs, E4 and CP33. In summary, the presence of IgG2b MAbs directed to surface components of C. trachomatis at certain times during the course of infection may play a role in enhancing the infectivity of this pathogen. PMID- 9199439 TI - Role of bacterial Mn-cofactored superoxide dismutase in oxidative stress responses, nasopharyngeal colonization, and sustained bacteremia caused by Haemophilus influenzae type b. AB - Haemophilus influenzae type b, a causative agent of bacterial sepsis and meningitis in young children, contains a single superoxide dismutase (SOD), a cytoplasmic MnSOD. To study the role of this enzyme, a chromosomal sodA::lacZ mutant (M-2) was constructed. M-2 had an increased sensitivity towards oxygen and the redox-active agent paraquat. A 3.4-fold increase in sodA-lacZ expression was found in M-2 grown with oxygen supply rates between 3 and 36 mmol of O2/liter/h. In similar experiments with the wild type, assaying SodA activity, a 3.1-fold increase was found. Both the wild type and M-2 grew best at the lowest oxygen supply rate tested, consistent with the notion that H. influenzae prefers a more anaerobic environment. In the infant rat model of infection, the ability of M-2 to colonize the nasopharynx was found to be impaired, but its ability to cause invasive disease was unaffected. This suggests that after invasion, the growth disadvantage imposed by a SodA- phenotype is not limiting. PMID- 9199440 TI - Ribotypes and virulence gene polymorphisms suggest three distinct Listeria monocytogenes lineages with differences in pathogenic potential. AB - A total of 133 Listeria monocytogenes isolates were characterized by ribotyping and allelic analysis of the virulence genes hly, actA, and inlA to uncover linkages between independent phylogenetic and specific virulence markers. PCR restriction fragment length polymorphisms revealed 8 hly, 11 inl4, and 2 actA alleles. The combination of these virulence gene alleles and ribotype patterns separated L. monocytogenes into three distinct lineages. While distinct hly and inlA alleles were generally found to cluster into these three lineages, actA alleles segregated independently. These three phylogenetic lineages were confirmed when 22 partial actA DNA sequences were analyzed. The clinical history of the L. monocytogenes strains showed evidence for differences in pathogenic potential among the three lineages. Lineage I contains all strains isolated during epidemic outbreaks of listeriosis, while no human isolates were found in lineage III. Animal isolates were found in all three lineages. We found evidence that isolates from lineages I and III have a higher plaquing efficiency than lineage II strains in a cell culture assay. Strains from lineage III also seem to form larger plaques than strains from lineage II. A distinctive ribotype fragment and unique 16S rRNA gene sequences furthermore suggest that lineage III might represent a L. monocytogenes subspecies. None of the 20 human isolates available but 11% of our animal isolates were grouped in this lineage, indicating that strains in this lineage might have reduced virulence for humans. PMID- 9199441 TI - Sialic acid-dependent recognition of laminin and fibrinogen by Aspergillus fumigatus conidia. AB - In an attempt to define the molecular basis of the adherence of Aspergillus fumigatus conidia to the host tissues, a step which might be mediated by the recognition of basement membrane laminin or fibrinogen, we analyzed the binding of these glycoproteins by flow cytometry and a microtiter plate adherence assay. Flow cytometry revealed that the binding of fluorescein isothiocyanate-labeled laminin to conidia was saturable and specific. Moreover, the ability of conidia to bind laminin increased with their maturation. Competition experiments showed a cross-reactivity between laminin and fibrinogen binding and a lack of interactions with glycosaminoglycans. In addition, the binding of laminin was not inhibited by the different adhesive synthetic peptides tested. Furthermore, the microtiter plate assay of adherence to chymotrypsin degradation products of laminin or fibrinogen purified by gel filtration suggested a unique binding site common to sequential degradation fragments or the presence of multiple binding sites on the two ligands. Therefore, the role of carbohydrates in the recognition process was investigated. Among the carbohydrates tested, constitutive of the conidial wall or of the oligosaccharide side chains of laminin and fibrinogen, only N-acetylneuraminic acid and sialyllactose inhibited the binding of these glycoproteins to conidia. In conclusion, these results strengthen the idea that the laminin and fibrinogen receptors in A. fumigatus are identical and suggest an interaction mediated by a sialic acid-specific lectin of the conidial wall. PMID- 9199442 TI - Effects of neutrophil, natural killer cell, and macrophage depletion on murine Clostridium piliforme infection. AB - Clostridium piliforme infection (Tyzzer's disease) induces enterohepatic disease in many domestic and laboratory animals. Murine susceptibility to Tyzzer's disease varies with host strain, age, and immune status However, little is known about the role of the immune system in control of this disease. To investigate the role of host immunity in Tyzzer's disease, mice were depleted of either neutrophils, natural killer cells, or macrophages by antibody administration or chemotherapy. After depletion, DBA/2 mice, which are naturally susceptible to C. piliforme, or naturally resistant C57BL/6 mice were inoculated intravenously with C. piliforme. Animals were euthanized 3 days postinoculation and evaluated for gross and histologic lesions and hepatic bacterial load. In juvenile DBA/2 or C57BL/6 mice, depletion of either neutrophils or natural killer cells increased severity of disease. In adult mice, depletion of natural killer cells significantly increased severity of Tyzzer's disease in the resistant (C57BL/6) but not in the susceptible (DBA/2) strain. Macrophage depletion did not alter the course of infection in either mouse strain. These studies indicate an important role for neutrophils and natural killer cells in the pathogenesis of murine Tyzzer's disease. The role of macrophages in murine C. piliforme infection will require further evaluation. PMID- 9199443 TI - Cytostatic and cytotoxic effects of activated macrophages and nitric oxide donors on Brugia malayi. AB - The susceptibility of Brugia malayi microfilariae and adults to injury by the murine macrophage cell line J774 activated with gamma interferon and bacterial lipopolysaccharide has been examined in vitro. Parasites of both stages showed a decline in viability over 48 h of coculture with activated macrophages, assessed by their capacity to reduce the tetrazolium salt 3-[4,5-diethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT), although adult parasites were more resistant than microfilariae. Removal of parasites to cell-free medium following exposure to activated macrophages for up to 48 h resulted in partial recovery of their capacity to reduce MTT, suggesting that the effects were primarily cytostatic. However, prolonged exposure to activated J774 cells for 72 h resulted in parasite death. Addition of the nitric oxide synthase inhibitor L-NMMA (N(G)-monomethyl-L arginine monoacetate) indicated that nitric oxide derivatives were responsible for cytostasis and ultimate toxicity. The toxicity of nitric oxide derivatives was confirmed by coincubation of parasites with chemical donors, although far higher concentrations were required than those generated by activated J774 cells, implying additional complexity in macrophage-mediated cytotoxicity. These experiments further suggested that peroxynitrite or its by-products were more potently damaging to filariae than nitric oxide per se. Examination of ultrastructural changes on exposure of parasites to activated macrophages or donors of nitric oxide indicated that hypodermal mitochondria were highly vacuolated, with less prominent cristae. The data are discussed with reference to immunity to lymphatic filariae and their mechanisms of energy generation. PMID- 9199444 TI - Clostridium difficile toxin A induces the release of neutrophil chemotactic factors from rat peritoneal macrophages: role of interleukin-1beta, tumor necrosis factor alpha, and leukotrienes. AB - Clostridium difficile produces a potent enterotoxin and cytotoxin, toxins A and B, respectively, which appear to be responsible for pseudomenbranous colitis and antibiotic-associated diarrhea. In the present study we explored the neutrophil migration evoked by toxin A in the peritoneal cavities and subcutaneous air pouches of rats and examined the role of macrophages and their inflammatory mediators in this process. Toxin A causes a significant dose-dependent neutrophil influx into the peritoneal cavity, with a maximal response at 0.1 microg/ml and at 4 h. The depletion of macrophages by peritoneal washing prevents the toxin A induced neutrophil migration into the peritoneal cavity. In contrast, an increase in macrophages induced by peritoneal injection of thioglycolate amplifies this toxin effect on neutrophil migration. Furthermore, the injection of supernatants from toxin A-stimulated macrophages into the rat peritoneal cavity causes significant neutrophil migration. Pretreatment of rats with BWA4C, nordihydroguaiaretic acid, mepacrine, or dexamethasone inhibits the neutrophil migration evoked by toxin A in the peritoneal cavities. However, pretreatment with the cyclooxygenase inhibitor indomethacin or the platelet-activating factor antagonist BN52021 fails to alter toxin A-induced neutrophil migration. Toxin A was also injected into air pouches of normal rats or rats pretreated with anti interleukin-1beta (anti-IL-1beta) or anti-tumor necrosis factor alpha (anti-TNF alpha) antibodies. Anti-TNF-alpha or anti-IL-1beta antibodies significantly reduce the neutrophil migration induced by toxin A. These data suggest that neutrophil migration evoked by toxin A is in part dependent on macrophage-derived cytokines, such as TNF-alpha and IL-1beta, and leukotrienes. These mediators may help to explain the intense inflammatory colitis caused by C. dificile toxin A in an experimental animal model of this disease. PMID- 9199445 TI - Pseudomonas aeruginosa lipopolysaccharide binds galectin-3 and other human corneal epithelial proteins. AB - The aim of this study was to test whether galectin-3 is present in human corneal epithelium and whether lipopolysaccharide (LPS) purified from Pseudomonas aeruginosa ATCC 19660 binds to this animal lectin and/or to another human corneal epithelial protein(s) (HCEP) and to confirm which component of LPS (inner or outer core or lipid A) is important in bacterial binding by using the eye in organ culture. LPS isolated and purified from P. aeruginosa ATCC 19660 and a commercial LPS (serotype 10) differed in polyacrylamide gel analysis but bound similarly to blotted HCEP. Binding was determined to be a receptor-ligand type of interaction by the solid-phase assay, because it was both specific and saturable. Several LPS binding proteins in HCEP were identified by an overlay method. Western blotting with antibody against galectin-3 revealed the presence of this protein in both freshly isolated and cultured transformed human corneal epithelium. Binding inhibition assays showed that antibody specific for the outer core region of LPS and an anti-galectin antibody significantly inhibited bacterial binding in vitro. These data provide further evidence that LPS is an important adhesin of P. aeruginosa, that it binds to protein receptor molecules in HCEP, that one of the LPS binding proteins is galectin-3, and that the outer core portion of the molecule appears to be critical for LPS binding to the eye. PMID- 9199446 TI - Proinflammatory cytokine deficiency and pathogenesis of Pseudomonas aeruginosa keratitis in aged mice. AB - Corneal clarity in young adult Swiss (HSD:ICR) mice is restored after Pseudomonas aeruginosa infection. Previous data showed that this response involves a rapid up regulation of constitutive intercellular cell adhesion molecule-1 (ICAM-1) and migration of inflammatory cells into the cornea. In contrast, in aged mice, there is no up-regulation of corneal ICAM-1, inflammatory cell infiltration into the cornea is delayed, and the cornea perforates. Therefore, the aim of this study was to test whether specific cytokines which up-regulate ICAM-1 expression differ in young and aged mice. Corneas of young (6- to 8-week-old) and aged (1- to 2 year-old) mice were scarified and inoculated with P. aeruginosa. The eyes were graded for pathologic changes (score 0 to +4); at 6, 12, 24, and 48 h postinfection (p.i.), six mice from each age group were sacrificed. Three corneas from each respective group were excised for quantitation of interleukin-1beta (IL 1beta), tumor necrosis factor alpha, and gamma interferon (IFN-gamma) by enzyme linked immunosorbent assay. The remaining three corneas from each age group were harvested for quantitation of viable bacteria by direct plate count determination and for infiltrating polymorphonuclear leukocytes (PMNs) by a myeloperoxidase (MPO) assay. Compared to those of young mice, the corneas of infected aged mice had less IL-1beta at 6 h p.i. (P < or = 0.04) and less IFN-gamma at 12 to 48 h p.i. (P < or = 0.05). Also, compared to those of young mice, corneas of aged mice had fewer PMNs (P < or = 0.008) by the MPO assay at 6 h p.i. and more viable bacteria (P < or = 0.01) per cornea by plate count determination at 24 h p.i. These data suggest that the lack of up-regulation of ocular ICAM-1 in aged mice may reflect a reduction in both IL-1beta and IFN-gamma levels in the infected cornea. Consequently, a sufficient number of PMNs and other inflammatory cells fail to rapidly migrate into the infected corneas of aged mice, the bacterial load is initially greater than that in young mice, and the cornea perforates. PMID- 9199447 TI - High-frequency invasion of epithelial cells by Streptococcus pyogenes can be activated by fibrinogen and peptides containing the sequence RGD. AB - The ability of Streptococcus pyogenes to invade human epithelial cells has been suggested to be an important contributing factor to the bacterium's ability to cause severe, invasive infections. We know little, however, of the mechanism underlying intracellular invasion by this organism. In this study, we demonstrate that the invasion of cultured human epithelial cells by a serotype M1 strain of S. pyogenes (strain 90-226) is stimulated over 50-fold by the addition of fetal calf serum (FCS) to the cell culture medium (RPMI medium). Purified human fibrinogen and peptides containing the sequence Arg-Gly-Asp (RGD) were also found to promote bacterial invasion of cultured cells. Experiments that demonstrate that the agonists stimulate invasion by interacting with bacterial cells are described. Invasion stimulation did not appear to involve de novo synthesis of a bacterial protein, as FCS and fibrinogen stimulated invasion in the presence of chloramphenicol. Although the agonists stimulated adherence by up to threefold, strain 90-226 efficiently adhered to cultured cells in unsupplemented RPMI medium. The invasion index (the number of internalized CFU/the number of adherent CFU) of strain 90-226 was increased 10- to 25-fold by the addition of the agonists. Postinternalization survival of bacteria was unaffected by fibrinogen or FCS. Thus, the agonistic factors affect the efficiency by which adherent bacteria are internalized by epithelial cells. PMID- 9199448 TI - Cloning and characterization of a Prevotella melaninogenica hemolysin. AB - Hemolysins have been proven to be important virulence factors in many medically relevant pathogenic organisms. Their production has also been implicated in the etiology of periodontal disease. Hemolytic strain 361B of Prevotella melaninogenica, a putative etiologic agent of periodontal disease, was used in this study. The cloning, sequencing, and characterization of phyA, the structural gene for a P. melaninogenica hemolysin, is described. No extensive sequence homology could be identified between phyA and any reported sequence at either the nucleotide or amino acid level. As predicted from sequence analysis, this gene produces a 39-kDa protein which has hemolytic activity as measured by zymogram analysis. Unlike many Ca2+-dependent bacterial hemolysins, both the cloned and native PhyA proteins were enhanced by the presence of EDTA in a dose-dependent fashion with 40 mM EDTA allowing maximum activity. Ca2+ and Mg2+ were found to be inhibitory. The hemolytic activity also was found to have a dose-dependent endpoint. Through recovery of hemolytic activity from a spent reaction, this endpoint was shown to be the result of end product inhibition. This is the first report describing the cloning and sequencing of a gene from P. melaninogenica. PMID- 9199449 TI - Expression and serologic activity of a soluble recombinant Plasmodium vivax Duffy binding protein. AB - Plasmodium vivax Duffy binding protein (DBP) is a conserved functionally important protein. P. vivax DBP is an asexual blood-stage malaria vaccine candidate because adhesion of P. vivax DBP to its erythrocyte receptor is essential for the parasite to continue development in human blood. We developed a soluble recombinant protein of P. vivax DBP (rDBP) and examined serologic activity to it in residents of a region of high endemicity. This soluble rDBP product contained the cysteine-rich ligand domain and most of the contiguous proline-rich hydrophilic region. rDBP was expressed as a glutathione S transferase (GST) fusion protein and was isolated from GST by thrombin treatment of the purified fusion protein bound on glutathione agarose beads. P. vivax rDBP was immunogenic in rabbits and induced antibodies that reacted with P. vivax and Plasmodium knowlesi merozoites. Human sera from adult residents of a region of Papua New Guinea where malaria is highly endemic or P. vivax-infected North American residents reacted with rDBP in an immunoblot and an enzyme-linked immunosorbent assay. The reactivity to reduced, denatured P. vivax rDBP and the cross-reactivity with P. knowlesi indicated the presence of immunogenic conserved linear B-cell epitopes. A more extensive serologic survey of Papua New Guinea residents showed that antibody response to P. vivax DBP is common and increases with age, suggesting a possible boosting of the antibody response in some by repeated exposure to P. vivax. A positive humoral response to P. vivax DBP correlated with a significantly higher response to P. vivax MSP-1(19). The natural immunogenicity of this DBP should strengthen its usefulness as a vaccine. PMID- 9199450 TI - Utilization of iron-catecholamine complexes involving ferric reductase activity in Listeria monocytogenes. AB - Listeria monocytogenes is a ubiquitous potentially pathogenic organism requiring iron for growth and virulence. Although it does not produce siderophores, L. monocytogenes is able to obtain iron by using either exogenous siderophores produced by various microorganisms or natural catechol compounds widespread in the environment. In the presence of tropolone, an iron-chelating agent, growth of L. monocytogenes is completely inhibited. However, the growth inhibition can be relieved by the addition of dopamine or norepinephrine under their different isomeric forms, while the catecholamine derivatives 4-hydroxy-3 methoxyphenylglycol and normetanephrine did not relieve the inhibitory effect of tropolone. Preincubation of L. monocytogenes with chlorpromazine and yohimbine did not antagonize the growth-promoting effect of catecholamines in iron complexed medium. In addition, norepinephrine stimulated the growth-promoting effect induced by human transferrin in iron-limited medium. Furthermore, dopamine and norepinephrine allowed 55Fe uptake by iron-deprived bacterial cells. The uptake of iron was energy dependent, as indicated by inhibition of 55Fe uptake at 0 degrees C as well as by preincubating the bacteria with KCN. Inhibition of 55Fe uptake by L. monocytogenes was also observed in the presence of Pt(II). Moreover, when assessed by a whole-cell ferric reductase assay, reductase activity of L. monocytogenes was inhibited by Pt(II). These data demonstrate that dopamine and norepinephrine can function as siderophore-like compounds in L. monocytogenes owing to their ortho-diphenol function and that catecholamine-mediated iron acquisition does not involve specific catecholamine receptors but acts through a cell-bound ferrireductase activity. PMID- 9199451 TI - Rickettsia rickettsii infection of cultured human endothelial cells induces NF kappaB activation. AB - Rickettsia rickettsii, the etiologic agent of Rocky Mountain spotted fever, is an obligate intracellular bacterial organism that infects primarily the vascular endothelial cells (EC). A component of the EC response to infection is transcriptional activation, which may contribute to the thrombotic and inflammatory consequences of disease. In this study, we explore R. rickettsii induced activation of the nuclear factor-kappaB/Rel (NF-kappaB) family of transcription factors involved in early transcriptional responses to injurious stimuli. Two NF-kappaB species were activated by infection and reacted with a double-stranded oligonucleotide probe corresponding to the kappaB binding domain of the murine kappa light-chain gene enhancer. Gel supershift analysis demonstrated the reactivity of these complexes with antibodies against p65 and p50, and the induced species were tentatively identified as p50-p50 homodimers and p50-p65 heterodimers. Semiquantitative reverse transcription-PCR analysis revealed dramatic increases in the steady-state levels of mRNA coding for the inhibitory subunit of NF-kappaB (IkappaB alpha), transcription of which is enhanced by the binding of NF-kappaB within the IkappaB alpha promoter region. NF kappaB activation was first detected 1.5 h following infection and was biphasic, with an early peak of activation at approximately 3 h, a return to baseline levels at 14 h, and even higher levels of activation at 24 h. It is likely that NF-kappaB activation requires cellular uptake of R. rickettsii, since treatment of EC with cytochalasin B during infection to block entry inhibited activation by only 70% at 3 h. R. rickettsii-induced activation of NF-kappaB may be an important controlling factor in the transcriptional responses of EC to infection with this obligate intracellular organism. PMID- 9199452 TI - The phagosomal environment protects virulent Mycobacterium avium from killing and destruction by clarithromycin. AB - Murine bone marrow-derived macrophages (Mphis) infected with virulent strains of Mycobacterium avium (TMC 724 and a human clinical isolate) or with an avirulent opaque variant that spontaneously dissociates from the virulent human clinical isolate were subjected to a prolonged and continuous treatment with clarithromycin added at the MIC. The efficiency of this antibiotic in terms of inhibition of bacterial growth and bacterial degradation was evaluated during a 21-day treatment period. Growth was assessed by determination of CFU of intracellular bacteria and by a quantitative ultrastructural analysis which allowed us also to determine the extent of bacterial degradation. A similar treatment was applied to the same strains growing in liquid medium. Our data show that in liquid medium, clarithromycin caused a 90% decrease in CFU within 7 days of treatment. When applied to Mphis infected with virulent M. avium, clarithromycin immediately arrested bacterial growth but was unable to fully kill and degrade intracellularly growing virulent bacteria. After 21 days of treatment, 25% of intracellular bacteria were still morphologically intact. These bacteria resumed growth upon removal of the antibiotic, with a normal replication rate. These bacteria had not become more resistant to the drug, since the MIC was unchanged as compared to the one determined for the initial stock used to infect Mphis. Our data therefore suggest that the intraphagosomal environment protects bacteria from degradation. We propose that the inability of the drug to completely destroy bacteria is the result of a limited accessibility of the drug due to prevention of fusions between the immature phagosomes in which virulent bacteria reside and lysosomes in which clarithromycin accumulates. In accord with our proposal, we show that the avirulent opaque variant, which does not prevent phagosome-lysosome fusions (unpublished data), is finally destroyed by clarithromycin even within the phagosomal environment. PMID- 9199453 TI - A synthetic lipopolysaccharide-binding peptide based on the neutrophil-derived protein CAP37 prevents endotoxin-induced responses in conscious rats. AB - The lipid A component of lipopolysaccharide (LPS) derived from Escherichia coli has been implicated as a significant mediator in the development of circulatory and metabolic dysfunction and lethality associated with sepsis. A synthetic peptide corresponding to amino acid residues 20 through 44 of the neutrophil derived 37-kDa cationic antimicrobial protein (CAP37 P(20-44)) possesses lipid A binding characteristics which may be useful in attenuating in vivo responses induced during circumstances of endotoxemia, including sepsis. The E. coli LPS to be used in the in vivo study was shown to be attenuated by CAP37 P(20-44) in a dose-dependent manner in the in vitro reaction with Limulus amoebocyte lysate. Intravenous infusion of CAP37 P(20-44) (1.5 or 3.0 mg/kg of body weight) with E. coli LPS (250 microg/kg over 30 min) into conscious, unrestrained rats prevented LPS-induced hyperdynamic and hypodynamic circulatory shock, hyperlactacidemia, and leukopenia in a dose-related fashion. CAP37 P(20-44) (0.2, 1.0, and 5.0 mg/kg) administered intravenously to conscious, actinomycin D-sensitized rats following a lethal dose of LPS neutralized LPS toxicity, resulting in dose dependent 7-day survival rates of 30, 50, and 80%, respectively. CAP37 P(20-44) (5.0 mg/kg) significantly inhibited the endotoxin-induced increase in circulating tumor necrosis factor alpha in sensitized rats. These data demonstrate that CAP37 P(20-44) has the capacity to abolish in vivo biological responses to LPS that are relevant to human sepsis and to significantly neutralize the toxicity of circulating E. coli LPS. PMID- 9199454 TI - Pathogenicity island sequences of pyelonephritogenic Escherichia coli CFT073 are associated with virulent uropathogenic strains. AB - Urinary tract infection is the most frequently diagnosed kidney and urologic disease, and Escherichia coli is by far the most common etiologic agent. Defined blocks of DNA termed pathogenicity islands have been found in uropathogenic strains to carry genes not generally found in fecal strains. We have identified one of these regions of DNA within the chromosome of the highly virulent E. coli CFT073, isolated from the blood and urine of a woman with acute pyelonephritis. This strain, which is cytotoxic for cultured renal cells and causes acute pyelonephritis in transurethrally infected CBA mice, contains two distinct copies of the pap operon and is hemolytic. One pap operon was localized on a cosmid clone which was used to identify three overlapping cosmid clones. By using restriction mapping, DNA hybridization, sequencing, and PCR amplification, a region of approximately 50 kb was found to be present in this uropathogenic strain and to have no corresponding sequences in E. coli K-12. This gene block also carries hemolysin genes hlyCABD. The pathogenicity island begins 7 bp downstream of dadX (catabolic alanine racemase; 26.55 min) and ends at a position in the K-12 genome 75 bp downstream of the metV tRNA gene (62.74 min); this suggests that a chromosomal rearrangement has occurred relative to the K-12 linkage map. The junctions of the pathogenicity island were verified by PCR amplification directly from the genomic DNA of strain CFT073. DNA sequencing within the boundaries of the junctions revealed genes not previously identified in E. coli or in some cases bearing no known homologs. When used as probes for DNA hybridization, these sequences were found significantly more often in strains associated with the clinical syndromes of cystitis (82%) and acute pyelonephritis (79%) than in fecal strains (19%; P < 0.001). PMID- 9199455 TI - Intranasal immunogenicity and adjuvanticity of site-directed mutant derivatives of cholera toxin. AB - Genetically modified derivatives of cholera toxin (CT), harboring a single amino acid substitution in and around the NAD binding cleft of the A subunit, were isolated following site-directed mutagenesis of the ctxA gene. Two mutants of CT, designated CTS106 (with a proline-to-serine change at position 106) and CTK63 (with a serine-to-lysine change at position 63), were found to have substantially reduced ADP-ribosyltransferase activity and toxicity; CTK63 was completely nontoxic in all assays, whereas CTS106 was 10(4) times less toxic than wild-type CT. The mucosal adjuvanticity and immunogenicity of derivatives of CT were assessed by intranasal immunization of mice, with either ovalbumin or fragment C of tetanus toxin as a bystander antigen. Mice immunized with wild-type CT produced both local (immunoglobulin A in mucosal washes) and systemic immune responses to both CT and bystander antigens. CTS106 showed good local and systemic responses to bystander proteins and to itself. Interestingly, mice immunized with the nontoxic derivative of CT, CTK63, generated weak immune responses to the bystander antigens which were similar to those achieved when CT B subunit was used as an adjuvant. In parallel experiments, an equivalent nontoxic mutant of the Escherichia coli heat-labile enterotoxin, LTK63 (with a serine-to-lysine change at position 63), was tested (9). In contrast to CTK63, LTK63 was found to be more immunogenic and a better intranasal adjuvant than recombinant heat-labile enterotoxin B subunit or CTK63. This information, together with data on immunoglobulin subclass responses, suggests that although highly homologous, CT and heat-labile enterotoxin should not be considered biologically identical in terms of their ability to act as intranasal adjuvants. PMID- 9199456 TI - Macrophages and enriched populations of T lymphocytes interact synergistically for the induction of severe, destructive Lyme arthritis. AB - Hamsters receiving both macrophages exposed to Formalin-inactivated Borrelia burgdorferi (Mphi-FBb) and enriched populations of either immune or naive T lymphocytes developed severe swelling of the hind paws when infected with B. burgdorferi. Swelling was detected 6 days after infection, peaked on day 10, and gradually decreased. Swelling was also observed in the hind paws of hamsters infused with only Mphi-FBb or only enriched populations of either immune or naive T lymphocytes after infection with B. burgdorferi. However, the swelling detected in these hamsters was less severe and of shorter duration. In addition, hamsters receiving both macrophages not exposed to Formalin-inactivated B. burgdorferi (Mphi-NFBb) and enriched populations of either immune or naive T lymphocytes failed to develop severe swelling after infection with B. burgdorferi. No swelling was also observed in hamsters infused with both Mphi-FBb and enriched populations of immune T lymphocytes and then inoculated with spirochetal growth medium. We further showed that macrophages and enriched populations of T lymphocytes did not interact synergistically for controlling B. burgdorferi infection, as spirochetes were readily recovered from the tissues of all cell transfer recipients infected with B. burgdorferi. These findings demonstrate that hamsters infused with both Mphi-FBb and enriched populations of either immune or naive T lymphocytes develop a more fulminate arthritis after infection with B. burgdorferi than recipients infused with either cell type alone. These findings suggest that macrophages and T lymphocytes interact synergistically for the induction of severe, destructive Lyme arthritis. PMID- 9199457 TI - An in vitro model for infection with Leishmania major that mimics the immune response in mice. AB - By using a primary in vitro response specific for Leishmania major, normal T cells from resistant CBA/CaH-T6J and susceptible BALB/c mice commit to a Th1 and a Th2 response, respectively. Since commitment occurred, we measured the production of gamma interferon (IFN-gamma), interleukin-1 (IL-1), IL-2, IL-4, IL 5, IL-10, and IL-12, prostaglandin E2 (PGE2), transforming growth factor beta (TGF-beta), and nitric oxide in the first 7 days of the response to identify factors that are critical for Th1 and Th2 development. While cells from resistant CBA mice produced more IFN-gamma, IL-10, and nitric oxide, cells from susceptible BALB/c mice produced more IL-1alpha, IL-5, PGE2, and TGF-beta. Although substantial amounts of IL-12 were detected, IL-12 did not associate with either Th1 or Th2 development. We did not anticipate that cells from resistant CBA mice would make more IL-10 in vitro. However, this also occurred in vivo since CBA mice produced substantial amounts of IL-10 following infection with L. major. Moreover, adding anti-IL-10 to primary in vitro responses enhanced production of IFN-gamma and nitric oxide by cells from CBA and BALB/c mice. Therefore, IL-10 cannot be regarded as a cytokine that associates with susceptibility to infection with L. major. Finally, the data presented here suggest that a collection of factors that can be produced by accessory cells influence Th commitment (e.g., IL 1, PGE2, and TGF-beta favor Th2 development). PMID- 9199458 TI - Protective responses against skin-dwelling microfilariae of Onchocerca lienalis in severe combined immunodeficient mice. AB - Inoculation of severe combined immunodeficient (SCID) mice with microfilariae of Onchocerca lienalis results in a sustained infection of the skin, extending for months beyond the point at which the parasites are eliminated from immunocompetent BALB/c controls. Reconstitution of SCID mice with spleen cells, thymocytes, or CD4+-cell-enriched splenocytes from naive BALB/c donors confers the ability to mount a protective immune response, leading to the rapid elimination of microfilariae. High levels of interleukin-5 and low levels of gamma interferon in the sera of reconstituted SCID mice during the destruction of microfilariae suggest that this protective immune response is directed by Th2 lymphocytes, mirroring that observed in immunocompetent controls. Unexpectedly, abbreviation of primary infections of unreconstituted SCID mice with the drug ivermectin induces resistance to reinfection with microfilariae at a level equivalent to that induced in secondarily infected, immunocompetent controls. In contrast to protection mediated by adoptive reconstitution, resistance induced by ivermectin-abbreviated infection occurs in the absence of T cells and in association with negligible levels of serum interleukin-5 and gamma interferon. This points to the activation of some alternative host defense mechanism that operates after the clearance of therapeutic levels of drug. Such a response could have important implications for the treatment of human onchocerciasis and may go some way in explaining the long-term suppression of microfilariae observed in patients after treatment with ivermectin. PMID- 9199459 TI - Identification and characterization of S fimbria-binding sialoglycoproteins on brain microvascular endothelial cells. AB - We have previously shown that S-fimbriated Escherichia coli binds brain microvascular endothelial cells (BMEC) via a lectin-like activity of SfaS adhesin specific for NeuAc alpha2,3-galactose; however, BMEC molecules bearing these epitopes have not been identified. In the present study, we showed that the expression of S fimbriae conferred a three-fold increase in adhesion of E. coli to cow, human, and rat BMEC but did not enhance E. coli adhesion to systemic vascular endothelial cells such as human umbilical vein endothelial cells and human aortic arterial endothelial cells. Two BMEC-binding molecules for S fimbriae were identified as 65 (major)- and 130 (minor)-kDa sialoglycoproteins by S fimbria immunoblotting and were purified from bovine BMEC by wheat germ agglutinin and Maackia amurensis lectin (specific to NeuAc alpha2,3-galactose) affinity chromatography. The 65-kDa BMEC glycoprotein showed effective inhibition of S fimbria-mediated binding of E. coli to BMEC. Polyclonal antibodies raised against the mixture of 65- and 130-kDa proteins reacted to 65-kDa protein present only on BMEC, not on systemic vascular endothelial cells. Immunoprecipitation of biotinylated BMEC membrane proteins and immunocytochemistry studies of BMEC with anti-S fimbria-binding protein antibodies revealed that the 65-kDa protein is a surface protein. The N-terminal amino acid sequence of 65- and 130-kDa proteins showed no significant sequence homology with any other known proteins. These findings suggest that 65- and 130-kDa proteins represent novel sialoglycoproteins involved in the binding of S-fimbriated E. coli to BMEC. PMID- 9199460 TI - Epithelial cell polarity affects susceptibility to Pseudomonas aeruginosa invasion and cytotoxicity. AB - Intact tissues are relatively resistant to Pseudomonas aeruginosa-induced disease, and injury predisposes tissue to infection. Intact epithelia contain polarized cells that have distinct apical and basolateral membranes with unique lipids and proteins. In this study, the role of cell polarity in epithelial cell susceptibility to P. aeruginosa virulence mechanisms was tested. Madin-Darby canine kidney (MDCK) cells, human corneal epithelial cells, and primary cultures of two different types of airway epithelial cells were grown on Transwell filters or in plastic tissue culture wells. P. aeruginosa invasion of cells was quantified by gentamicin survival assays with two isolates that invade epithelial cells (6294 and PAO1). Cytotoxic activity was assessed by trypan blue exclusion assays with two cytotoxic strains (6206 and PA103). Basolateral surfaces of cells were exposed by one of two methods: EGTA pretreatment of epithelial cells or growth of cells in low-calcium medium. Both methods of exposing basolateral membranes increased epithelial cell susceptibility to P. aeruginosa invasion and cytotoxicity. Migrating cells were also found to be more susceptible to P. aeruginosa invasion than confluent monolayers that had established membrane polarity. Monolayers of MDCK cells that had been selected for resistance to killing by concanavalin A were resistant to both cytotoxicity and invasion by P. aeruginosa because they were more efficiently polarized for their susceptibility to P. aeruginosa virulence factors than regular MDCK cells and not because they were defective in glycosylation. These results suggest that there are factors on the basolateral surfaces of epithelial cells that promote interaction with P. aeruginosa or that there are inhibitory factors on the apical cell surface. Thus, cell polarity of intact epithelia is likely to contribute to defense against P. aeruginosa infection. PMID- 9199461 TI - Analysis of toxicity of streptococcal pyrogenic exotoxin A mutants. AB - Streptococcal pyrogenic exotoxin A (SPE A) is secreted by some strains of Streptococcus pyogenes and is strongly associated with streptococcal toxic shock syndrome (STSS), a severe and often fatal illness. SPE A possesses a number of biological properties, some of which are shared with a group of exotoxins of streptococcal and staphylococcal origins, the pyrogenic toxin superantigens (PTSAgs). SPE A's most extensively studied property is superantigenicity. Superantigenic activation of T cells and monocytes stimulates the release of cytokines such as tumor necrosis factors alpha and beta, interleukin 1, and gamma interferon. These endogenous mediators are considered to be the primary cause of capillary leak, hypotension, and shock, the most severe manifestations of STSS. However, several studies have suggested that other properties of SPE A, such as ability to greatly enhance host susceptibility to endotoxin and ability to interact directly with endothelial cells, may play substantial roles in the syndrome. In this work we generated single- and double-site mutations of SPE A at residues K16, N20, C87, C90, C98, K157, S195, N20/C98, and N20/K157. The mutant SPE A's were analyzed in vivo for their lethal activity and in vitro for their superantigenic ability. Our results indicate that SPE A's ability to induce lethality and endotoxin enhancement does not require superantigenicity, and conversely superantigenicity does not necessarily lead to lethality. Thus, these properties and their relative contributions to the onset of hypotension and shock may be separable. Furthermore, evidence is presented that certain mutant toxins may be suitable for use as vaccine toxoids. PMID- 9199462 TI - Humoral and cellular immunity in secondary infection due to murine Chlamydia trachomatis. AB - A murine model of pneumonia due to the mouse pneumonitis agent (MoPn [murine Chlamydia trachomatis]) in mice deficient in CD4+ T-cell function (major histocompatibility complex [MHC] class II function [class II-/-], CD8+ T-cell function (beta2-microglobulin deficient, MHC class I deficient [Beta2m-/-]), B cell function (C57BL/10J-Igh(tm1Cgn) [Igh-/-]), and gamma interferon (IFN-gamma) (C57BL/6-Ifg(tm1) [Ifg-/-]) or interleukin-4 (C57BL/6J(tm1Cgn29) [IL4-/-]) production was employed to determine if each of these mechanisms was critical to resistance against reinfection by C. trachomatis or if alternate compensatory mechanisms existed in their absence which could potentially be exploited in vaccine development. Resistance to reinfection with MoPn was heavily dependent on CD4+ T cells. CD4 T-cell-deficient MHC class II-/- mice were very susceptible to reinfection with MoPn, showing the critical importance of this cell to resistance. These mice lacked antibody production but did produce IFN-gamma, apparently by mechanisms involving NK and CD8+ T cells. Neutralization of IFN gamma in these mice led to a borderline increase in susceptibility, showing a possible role (albeit small) of this cytokine in this setting. Tumor necrosis factor alpha (TNF-alpha) was also present at increased levels in these mice. Igh /- B-cell-deficient mice which produce no antibody to MoPn were only modestly more susceptible to reinfection than immunized B-cell-intact controls, showing that antibody, including lung immunoglobulin A, is not an absolute requirement for relatively successful host defense in this setting. Levels of lung IFN-gamma and TNF-alpha were elevated in Igh-/- mice compared to those in controls. IL-4-/- mice (deficient in Th2 function) could develop normal resistance to reinfection with MoPn. Conversely, normal mice rendered partially IFN-gamma deficient by antibody depletion were somewhat impaired in their ability to develop acquired immunity to MoPn, again indicating a role for this cytokine in host defense against rechallenge. Of most importance, however, congenitally IFN-gamma deficient Ifg-/- mice (which have elevated levels of other cytokines, including TNF-alpha and granulocyte-macrophage colony-stimulating factor) are paradoxically more resistant to MoPn rechallenge than controls, showing that IFN-gamma is not an absolute requirement for acquired resistance and implying the presence of very effective compensatory host defense mechanism(s). In vivo depletion of TNF-alpha significantly increased MoPn levels in the lungs in these mice. Thus, resistance to reinfection in this model is flexible and multifactorial and is heavily dependent on CD4+ T cells, with a probable role for IFN-gamma and TNF-alpha and a possible modest role for Th1-dependent antibody. Since IFN-gamma was dispensable in host defense, the highly effective mechanism or mechanisms which can compensate for its absence (which include TNF-alpha) deserve further study. PMID- 9199463 TI - Mycobacterium tuberculosis Des protein: an immunodominant target for the humoral response of tuberculous patients. AB - The phoA gene fusion methodology permitted the identification of a new Mycobacterium tuberculosis exported protein, Des. This protein has significant sequence similarities to plant acyl-acyl carrier protein desaturases, which are enzymes involved in general fatty acid biosynthesis as well as in mycolic acid biosynthesis in mycobacteria. As shown by Western blot and enzyme-linked immunosorbent assay experiments, the Des protein is a major B-cell antigen that was recognized by all the tuberculous M. tuberculosis- and M. bovis-infected human patients tested. PMID- 9199464 TI - Anti-Ehrlichia chaffeensis antibody complexed with E. chaffeensis induces potent proinflammatory cytokine mRNA expression in human monocytes through sustained reduction of IkappaB-alpha and activation of NF-kappaB. AB - Ehrlichia chaffeensis is an obligatory intracellular bacterium that infects monocytes and macrophages and is the etiologic agent of human ehrlichiosis in the United States. Our previous studies showed that the exposure of human monocytes to E. chaffeensis induces the expression of interleukin-1beta (IL-1beta), IL-8, and IL-10 genes in vitro but not the expression of tumor necrosis factor alpha (TNF-alpha) and IL-6 mRNAs. In this study, the effect of anti-E. chaffeensis antibody complexed with E. chaffeensis on the expression of major proinflammatory cytokines in human monocytes was examined. Human monocytic cell line THP-1 was treated with E. chaffeensis which had been preincubated with human anti-E. chaffeensis serum for 2 h, and the levels of cytokine mRNAs were evaluated by competitive reverse transcription-PCR. Anti-E. chaffeensis antibody complexed with E. chaffeensis significantly enhanced mRNA expression of IL-1beta in THP-1 cells. The expression of TNF-alpha and IL-6 mRNAs was also induced. The levels of secreted IL-1beta, TNF-alpha, and IL-6 during 24 h of stimulation were comparable to those induced by Escherichia coli lipopolysaccharide at 1 microg/ml. Fab fragment of anti-E. chaffeensis immunoglobulin G complexed with E. chaffeensis did not induce any of these three cytokines, indicating that ehrlichial binding is required for IL-1beta mRNA expression and that binding of the immune complex to the Fc gamma receptor is required for TNF-alpha and IL-6 mRNA expression and enhanced IL-1beta mRNA expression. Furthermore, prolonged degradation of IkappaB alpha and activation of NF-kappaB were demonstrated in THP-1 cells exposed to anti-E. chaffeensis serum and E. chaffeensis. This result implies that development of anti-E. chaffeensis antibody in patients can result in the production of major proinflammatory cytokines, which may play an important role in the pathophysiology of ehrlichiosis and immune responses to it. PMID- 9199465 TI - Effects of pH and salinity on the antimicrobial properties of clavanins. AB - Clavanins are histidine-rich, amidated alpha-helical antimicrobial peptides that were originally isolated from the leukocytes (hemocytes) of a tunicate, Styela clava. The activities of clavanin A amide and clavanin A acid against Escherichia coli, Listeria monocytogenes, and Candida albicans were substantially greater at pH 5.5 than at pH 7.4. In contrast, clavanin AK, a synthetic variant of clavanin A acid containing 4 histidine-->lysine substitutions exerted substantial activity at both pH 7.4 and pH 5.5. Each of these three clavanins permeabilized the outer and inner membranes of E. coli very effectively at pH 5.5, but only clavanin AK did so at pH 7.4. Unlike magainin 1 and cecropin P1, alpha-helical antimicrobial peptides from frog skin and porcine intestine, respectively, clavanins were broadly effective against gram-positive bacteria, including methicillin-resistant Staphylococcus aureus, as well as gram-negative organisms. Because clavanins exert substantial antimicrobial activity in 0.1 to 0.3 M NaCl, they provide templates for designing broad-spectrum peptide antibiotics intended to function in extracellular environments containing normal or elevated NaCl concentrations. The pH-dependent properties of histidine-rich antimicrobial peptides may allow the design of agents that would function selectively in acidic compartments, such as the gastric lumen, or within phagolysosomes. PMID- 9199466 TI - PlcR1 and PlcR2 are putative calcium-binding proteins required for secretion of the hemolytic phospholipase C of Pseudomonas aeruginosa. AB - The plcHR operon of Pseudomonas aeruginosa includes the structural gene for the hemolytic phospholipase C,plcH (previously known as plcS), and two overlapping, in-phase, genes designated plcR1 and plcR2. Hemolytic and phospholipase C (PLC) activities produced by Escherichia coli and P. aeruginosa T7 expression systems were measured in strains carrying both plcH and plcR genes and in strains carrying each gene separately. When plcH was expressed by itself in the E. coli T7 system, the area of the hemolytic zone on blood agar was less than twice the area of growth. By contrast, when plcR was coexpressed with plcH in this system, the area of the hemolytic zone was approximately 10 times that of the area of the growth on blood agar. Native polyacrylamide gel electrophoretic analyses of PlcH activity expressed in either the E. coli or the P. aeruginosa T7 system carrying plcH alone, or along with the plcR genes, suggest that PlcR either posttranslationally alters the physical or biochemical nature of PlcH or releases PlcH from a complex in the cell so that it can be secreted. The hypothesis that PlcR is involved in the secretion of PlcH is supported by the observation that the ratio of extracellular to cell-associated PlcH activity produced by P. aeruginosa strains containing an in-frame deletion in the chromosomal plcR genes is significantly reduced in comparison with this ratio seen with the wild-type parental strain. This defect in the secretion of PlcH can be complemented by the plcR genes in trans. Additional data suggest that PlcR does not directly affect the secretion of the nonhemolytic phospholipase C (PlcN). PlcR is highly similar to a calcium-binding protein (CAB) from Streptomyces erythraeus. PlcR and CAB contain typical motifs (EF hands) characteristic of eucaryotic calcium-binding proteins, including calmodulin. P. aeruginosa naturally produces membrane vesicles (MVs) containing extracellular proteins including PLC. MVs from the PAO1WT strain contained at least 10-fold more PLC specific activity than those isolated from a strain carrying a deletion of plcR (PAO1 deltaR). Immunogold electron microscopy of PAO1WT and PAO1 deltaR whole cells revealed a distribution of PlcH in these strains consistent with the hypothesis that PlcR is required for the secretion of PlcH. PMID- 9199467 TI - Differences in the association of Chlamydia trachomatis serovar E and serovar L2 with epithelial cells in vitro may reflect biological differences in vivo. AB - Chlamydia trachomatis serovar E is one of the most common bacterial sexually transmitted pathogens. Since it is an obligate intracellular bacterium, efficient colonization of genital mucosal epithelial cells is crucial to the infectious process. Serovar E elementary bodies (EB) metabolically radiolabeled with 35S-Cys Met and harvested from microcarrier bead cultures, which significantly improves the infectious EB-to-particle ratio, provided a more accurate picture of the parameters of attachment of EB to human endometrial epithelial cells (HEC-1B) than did less infectious 14C-EB harvested from flask cultures. Binding of serovar E EB was (i) equivalent at 35 and 4 degrees C, (ii) decreased by preexposure of EB to heat or the topical microbicide C31G, (iii) comparable among common eukaryotic cell lines (HeLa, McCoy), and (iv) significantly increased to the apical surfaces of polarized cells versus nonpolarized cells. In parallel experiments with C. trachomatis serovar L2, serovar E attachment was not affected by heparin or heparan sulfate whereas these glucosaminoglycans dramatically reduced serovar L2 attachment. These data were confirmed by competitive inhibition of serovar E binding and infectivity by excess unlabeled live and UV inactivated serovar E EB but not by excess serovar L2 EB. The noninvasive serovar E strains in the lumen of the genital tract enter and exit the apical domains of target columnar epithelial cells to spread canalicularly in an ascending fashion from the lower to the upper genital tract. In contrast, the invasive serovar L2 strains are primarily submucosal pathogens and likely use the glucosaminoglycans concentrated in the extracellular matrix to colonize the basolateral domains of mucosal epithelia to perpetuate the infectious process. PMID- 9199468 TI - Granuloma cytokines in murine cysticercosis. AB - Neurocysticercosis, caused by Taenia solium, is one of the most common causes of seizures worldwide. The symptoms result from granulomatous inflammation associated with dying cyst forms of the parasite. Although the invasive larvae can be killed by immune serum plus complement, immunity to the cyst stage depends on a cellular response. This dichotomous immune response is reminiscent of the extremes of the immune response associated with T helper 1 (Th1) and Th2 cytokine profiles. To characterize the cytokine response in cysticercosis, granulomas were removed from the peritoneal cavity of mice infected with Taenia crassiceps cysts and examined for cytokine message by in situ hybridization using 35S-labeled RNA probes. The granulomas were staged based on histologic appearance of the degenerating parasite. Message for gamma interferon (IFN-gamma) was identified by light microscopy in 11 of the 12 granulomas, and interleukin-2 (IL-2) message was identified in 9 of the 12. By laser scanning confocal microscopy, significantly increased IFN-gamma and IL-2 pixel intensity was identified in nearly all of the granulomas from early histologic stages. Message for IL-4 was seen in 6 of the 12 granulomas. Only granulomas with complete destruction of the parasite architecture displayed more than minimal amounts of IL-4 message by light microscopy, and only 2 of 12 granulomas had IL-4 pixel intensity significantly above background. Only minimal amounts of IL-10 message were detected in 4 of 11 granulomas. Thus, early granulomas in cysticercosis are predominantly associated with a Th1 response, whereas later granulomas, in which parasite destruction is complete, have a mixture of Th1 and IL-4. The Th1 response appears to play an important role both in the pathogenesis of disease as well as in the clearing of the parasites, with IL-4 involved in downregulation of the initial response. PMID- 9199469 TI - Induction of nitric oxide synthesis and xanthine oxidase and their roles in the antimicrobial mechanism against Salmonella typhimurium infection in mice. AB - The role of superoxide anion (O2-) and nitric oxide (NO) in the host defense mechanism against Salmonella typhimurium (LT-2) was examined by focusing on xanthine oxidase (XO) as an O2(-)-generating system and on inducible NO synthase (iNOS). When ICR mice were infected with a 0.1 50% lethal dose (2 x 10(5) CFU) of S. typhimurium, bacterial growth in the liver reached a peak value 3 days after infection (10(4.32) CFU/g of liver) and decreased thereafter. XO activity in the liver became maximum at 7 days after infection; the value was 34.6 +/- 1.4 mU/g of liver at 7 days (compared with 11.0 +/- 1.3 mU/g of liver before infection). The time profile of NO production in the liver as determined by electron spin resonance spectroscopy was consistent with that of XO activity. Histological examination of infected liver showed the formation of multiple microabscesses with granulomatous lesions consisting of polymorphonuclear cells and mononuclear cells, and iNOS-expressing cells were localized in the confined areas of the microabscesses. When XO inhibitors such as allopurinol and 4-amino-6 hydroxypyrazolo[3,4-d]pyrimidine (AHPP) were administered to the infected mice, the mortality of the mice was significantly increased (10 of 21 and 11 of 20 for the allopurinol- and AHPP-treated groups, respectively, versus 2 of 20 for control mice), and bacterial growth was significantly enhanced. A similar exacerbation of the infection was obtained with N(omega)-monomethyl-L-arginine (L NMMA) treatment of the mice. Of considerable importance is that granuloma formation in the liver was poorly developed by treatment with either XO inhibitors or L-NMMA. These results suggest that XO and NO play an important role in the antimicrobial mechanism against S. typhimurium in mice. PMID- 9199470 TI - Protective immunity against Clostridium difficile toxin A induced by oral immunization with a live, attenuated Vibrio cholerae vector strain. AB - Clostridium difficile causes pseudomembranous colitis through the action of Rho modifying proteins, toxins A and B. Antibodies directed against C. difficile toxin A prevent or limit C. difficile-induced colitis. We engineered plasmid pETR14, containing the hlyB and hlyD genes of the Escherichia coli hemolysin operon, to express a fusion protein containing 720 amino acid residues from the nontoxic, receptor-binding, carboxy terminus of C. difficile toxin A and the secretion signal of E. coli hemolysin A. We introduced pETR14 into Vibrio cholerae and found that the toxin A-HlyA fusion protein was secreted by a number of V. cholerae strains and recognized by both monoclonal and polyclonal anti-C. difficile toxin A antibodies. We introduced pETR14 into an attenuated V. cholerae strain, O395-NT, and inoculated rabbits orally with this construct. Colonization studies disclosed that the V. cholerae vector containing pETR14 was recoverable from rabbit ilea up to 5 days after oral inoculation. Vaccination produced significant systemic anti-C. difficile toxin A immunoglobulin G and anti-V. cholerae vibriocidal antibody responses. Vaccination also produced significant protection against toxin A in an ileal loop challenge assay, as assessed by determination of both fluid secretion and histological changes. These results suggest that the hemolysin system of E. coli can be used successfully in V. cholerae vector strains to effect secretion of large heterologous antigens and that a V. cholerae vector strain secreting a nontoxic, immunogenic portion of C. difficile toxin A fused to the secretion signal of E. coli HlyA induces protective systemic and mucosal immunity against this toxin. PMID- 9199471 TI - Invasion of cultured human epithelial cells by Klebsiella pneumoniae isolated from the urinary tract. AB - The mechanisms which enable entry into cultured human epithelial cells by Klebsiella pneumoniae were compared with those of Salmonella typhi Ty2. K. pneumoniae 3091, isolated from a urine sample of a patient with a urinary tract infection, invaded human epithelial cells from the bladder and ileocecum and persisted for days in vitro. Electron microscopic studies demonstrated that K. pneumoniae was always contained in endosomes. The internalization mechanism(s) triggered by K. pneumoniae was studied by invasion assays conducted with different inhibitors that act on prokaryotic and eukaryotic cell structures and processes. Chloramphenicol inhibition of bacterial uptake revealed that bacterial de novo protein synthesis was essential for efficient invasion by K. pneumoniae and S. typhi. Interference with receptor-mediated endocytosis by g-strophanthin or monodansylcadaverine and inhibition of endosome acidification by monensin reduced the number of viable intracellular K. pneumoniae cells, but not S. typhi cells. The depolymerization of microfilaments by cytochalasin D inhibited the uptake of both bacteria. Microtubule depolymerization caused by colchicine, demecolcine, or nocodazole and the stabilization of microtubules with taxol reduced only the invasion ability of K. pneumoniae. S. typhi invasion was unaffected by microtubule depolymerization or stabilization. These data suggest that the internalization mechanism triggered by K. pneumoniae 3091 is strikingly different from the solely microfilament-dependent invasion mechanism exhibited by many of the well-studied enteric bacteria, such as enteroinvasive Escherichia coli, Salmonella, Shigella, and Yersinia strains. PMID- 9199472 TI - Tyrosine phosphorylation is required for ehrlichial internalization and replication in P388D1 cells. AB - Replication of Ehrlichia risticii was inhibited in P388D1 cells when a protein tyrosine kinase inhibitor (genistein or herbimycin A) was added after internalization of the organism at 3 h postinfection. Upon addition of genistein at day 1, 2, 3, or 4 postinfection, further proliferation of E. risticii was prevented. The inhibition was reversible, since regrowth of E. risticii occurred upon the removal of genistein. Genistein prevented spreading of E. risticii from P388D1 cells to THP-1 cells. Genistein did not prevent binding of [35S]methionine labeled E. risticii to P388D1 cells but did prevent internalization of [35S]methionine-labeled E. risticii. 14CO2 production from L-[14C]glutamine in Percoll density gradient-purified E. risticii was not inhibited by genistein or herbimycin A, which suggests that these reagents did not directly inhibit ehrlichial energy metabolism. Double indirect immunofluorescence labeling with antiphosphotyrosine antibody and anti-E. risticii antibody revealed colocalization of tyrosine phosphoproteins with ehrlichial inclusions. There was, however, no colocalization of phosphotyrosine with phagosomes containing 0.5 microm-diameter fluorescent beads. Western immunoblot analysis revealed that 52- and 54-kDa proteins were tyrosine phosphorylated only in infected cells and that phosphorylation of these two proteins was reduced when infected cells were treated with genistein for 6 h. These results suggest that protein tyrosine phosphorylation is specific and essential for ehrlichial internalization, replication, and spreading in macrophages but not for binding. PMID- 9199473 TI - Characterization of a new isolate of Chlamydia trachomatis which lacks the common plasmid and has properties of biovar trachoma. AB - A Chlamydia trachomatis urethral isolate, alpha/95, yielding pgp3-negative but otherwise normal inclusions by immunofluorescence also gave negative results when pCT-homologous DNA was searched by PCR and Southern blotting. omp-1 sequence analysis identified alpha/95 as a new genotype B variant. These findings confirm that pCT is not required for chlamydial growth in vitro. PMID- 9199474 TI - A monoclonal antibody directed against the 97-kilodalton gonococcal hemin-binding protein inhibits hemin utilization by Neisseria gonorrhoeae. AB - Neisseria gonorrhoeae expresses two hemin-binding proteins (HmBPs) of 97,000 and 44,000 in molecular weight. A murine monoclonal antibody (MAb) produced against the 97-kDa HmBP from N. gonorrhoeae PID543 specifically inhibited in a concentration-dependent manner the ability of hemin to promote growth. The anti 97-kDa HmBP MAb competitively inhibited binding of the 97-kDa HmBP to a hemin agarose affinity column. In Western immunoblots, the MAb recognized the 97-kDa homologs from a limited survey of clinical gonococcal isolates. These results support the contention that the 97-kDa HmBP is involved in the gonococcal hemin acquisition pathway. PMID- 9199475 TI - Role of gamma interferon in the pathogenesis of bacteremic pneumococcal pneumonia. AB - Gamma interferon (IFN-gamma) concentrations in blood, but not in lungs, rose significantly at 24 to 48 h after murine pulmonary infection with virulent pneumococci. In contrast, infection with avirulent pneumococcal strains produced minimal rises in serum IFN-gamma concentrations. Compared with that of immunocompetent mice, mortality was appreciably increased after pulmonary infection of IFN-gamma gene knockout mice, suggesting a protective role for IFN gamma in host response to pneumococcal disease. PMID- 9199476 TI - Murine antibody responses to the verotoxin 1 B subunit: demonstration of major histocompatibility complex dependence and an immunodominant epitope involving phenylalanine 30. AB - Structurally conserved verotoxin 1 (VT1) mutant derivatives, showing reduced receptor binding and cytotoxicity, may serve as natural toxoids to protect against VT-mediated disease. In this study, the antibody responses to the wild type VT1 B subunit, a B-subunit mutant (Phe30Ala B), and the corresponding holotoxin (Phe30Ala HT) were examined in three inbred mouse strains. BALB/c (H 2d) and CBA (H-2k) mice produced strong antibody responses to both wild-type and mutant B subunits. VT1 B-raised sera reacted more strongly with VT1 B than with Phe30Ala B in enzyme-linked immunosorbent assays, while Phe30Ala B-raised sera reacted equally with VT1 B and Phe30Ala B. C57BL/6 (H-2b) and congenic BALB/c (BALB x B [H-2b]) mice produced no detectable antibody response to either VT1 B or Phe30Ala B. However, an anti-VT1 B antibody response was detected in H-2b mice immunized with biologically active Phe30Ala HT. Based on these observations, we conclude that the VT1 B subunit possesses a B-cell immunodominant epitope formed partly by phenylalanine 30 and that the B-subunit antibody response is dependent on the H-2 haplotype of the mouse strain. Our results also support a potential role for the A subunit in providing the T-cell help necessary to overcome a deficient B-subunit antibody response in H-2b mice. PMID- 9199478 TI - Lymphocyte proliferative responses of goats vaccinated with Brucella melitensis 16M or a delta purE201 strain. AB - The response to a Brucella melitensis purEK deletion mutant, delta purE201 (referred to as strain 201), was compared with the response to its parental strain, 16M, in juvenile goats. Proliferative responses to gamma-irradiated bacteria were detected earlier in strain 201-infected goats. Lymphocytes from strain 16M- or 201-infected goats proliferated in response to one-dimensional polyacrylamide gel electrophoresis-separated proteins of similar mass isolated from strain 16M or Brucella abortus RB51. Data from this study suggest that some antigens stimulating cell-mediated responses are conserved among Brucella species, as 201- and 16M-infected goats recognized similar proteins expressed by RB51 and 16M. PMID- 9199477 TI - Induction and characterization of heat shock proteins of Salmonella typhi and their reactivity with sera from patients with typhoid fever. AB - The heat shock protein (HSP) response of Salmonella typhi following exposure to elevated growth temperatures was studied. Three major proteins with molecular sizes of 58, 68, and 88 kDa were abundantly expressed when S. typhi cells were shifted from 37 to 45 degrees C and to 55 degrees C. These proteins were also constitutively expressed at 37 degrees C. Western blotting and immunoprecipitation studies with anti-HSP monoclonal antibodies revealed that the 58- and 68-kDa proteins were analogous to the GroEL and DnaK proteins, respectively, of Escherichia coli. These HSPs are also abundantly present in the outer membrane fraction of disrupted cells and, to a lesser extent, in the cytosol. Immunoblotting experiments with sera from patients with a culture positive diagnosis of typhoid fever showed the presence of antibodies to these HSPs. Nine of twelve sera reacted with the 58-, 68-, and 88-kDa proteins, while three sera reacted only with the 68- and 88-kDa proteins. All 10 sera from healthy individuals showed no binding to these HSPs. In light of the well documented roles of HSPs in the pathogenesis of microbial infections and as immunodominant antigens, these findings may be relevant for a better understanding of disease processes and for the future development of diagnostic and preventive strategies. PMID- 9199480 TI - Effects of anandamide on gestation in pregnant rats. AB - The present study was to test whether the recently described endogenous ligand for the cannabinoid receptor; arachidonyl-ethanolamide (anandamide, ANA), may produce similar effects on pregnancy as the main psychoactive component of marihuana: delta9-tetrahydrocannabinol (THC) in rats. ANA, THC (0.02 mg/kg i.p./day, respectively) or vehicle were injected daily over the third week of pregnancy. The pregnant rats were either killed on day 21 of pregnancy or followed up to delivery. Results show a significant increase in the duration of pregnancy after both THC and ANA treatment. Both drugs caused an increase in the frequency of stillbirths. The mothers' hormone contents in tissues and sera were measured. Decreased LH content was observed in the serum of treated animals. No changes in FSH content were observed either in the pituitary or in the sera. Pituitary prolactin (PRL) levels was lower in ANA treated animals as compared both to controls or THC treated subjects. The serum PRL content decreased in all experimental groups. Decrease in serum progesterone was more prominent in treated rats. Serum levels of prostaglandins (PGF 1alpha and PGF 2alpha) were significantly decreased after THC and ANA treatment. We conclude that ANA has the same tendency to change reproductory parameters in pregnant rats as THC, although in some cases the effects of ANA were slightly different from that of THC. Both endogenous and exogenous cannabinoids inhibit PG synthesis in pregnant rats and this maybe responsible for the delay constitute the mechanism in the onset of labour. PMID- 9199479 TI - FlaA, a putative flagellar outer sheath protein, is not an immunodominant antigen associated with Lyme disease. AB - FlaA was recently found to be associated with flagellar filaments of Borrelia burgdorferi. We tested whether antibodies to this protein are a good indicator of infection, as antibodies to FlaA proteins in other spirochetal infections show an increase in titer. Although overproduction of intact FlaA was highly toxic to Escherichia coli, truncated proteins which lacked the N-terminal signal sequence could be successfully overexpressed. Immunoblotting with sera from mammalian hosts infected with B. burgdorferi indicated that FlaA is not an immunodominant antigen in Lyme disease. However, sera from two patients reacted with both recombinant and native FlaA protein, suggesting that B. burgdorferi FlaA was antigenic and expressed in vivo. PMID- 9199481 TI - Distribution and disappearance of the radiolabeled carbon derived from L-arginine and taurine in the mouse. AB - L-arginine and taurine are still in the center of physiological and pharmacological research. Although the fate of nitrogen of both compounds and of the 35S-taurine is well-documented, the fate of the carbon skeleton has not been elucidated yet. We studied the organ distribution of 14C arginine and 14C taurine over time in the mouse using whole body autoradiography with densitometric image analysis. We describe different organ distribution patterns. Kidney, heart, lung, the Harderian gland, the central nervous system, intestine and testis showed a comparable pattern of arginine disappearance in contrast to rapid disappearance in the salivary gland and the accumulation pattern in bone and spleen. Data on 14C taurine of liver, kidneys, lung, testis and Harderian gland resembled the arginine pattern; Accumulation of taurine carbon was found in salivary gland, bone, intestine, heart and brain. Our studies challenge and demand further related studies to obtaining more information on the fate of the carbon skeleton of these amino acids. PMID- 9199482 TI - Inhibition of the rat adrenal ornithine decarboxylase activity by immobilization stress and/or dexamethasone. AB - The effects of immobilization stress and/or dexamethasone (DEX) on the adrenal ornithine decarboxylase (ODC) activities of sham-operated and adrenal medulloectomized (enucleated) male Sprague-Dawley rats were investigated. On day 11 after surgery, rats were injected with saline or DEX (1 mg/kg), 3 h before the time of sacrifice (0600 h or 1800 h). Four groups, from sham-operated and enucleated rats (ENU) treated with saline or DEX were subjected to immobilization stress for 1 h prior to sacrifice. Groups of rats from stress-sham-DEX, non stress-sham-DEX, stress-sham, non stress-sham, stress-ENU-DEX, non stress-ENU DEX, stress-ENU, and non stress-ENU were sacrificed at 0600 h or 1800 h on day 11 after surgery. Adrenal glands were excised and later analyzed for ODC activities. Results indicated that DEX and/or immobilization stress inhibited ODC activities (p<0.05) in normal and regenerating adrenal glands at 1800 h and ODC activity varies diurnally, the activity being greater at 1800 h than at 0600 hours (p<0.001). PMID- 9199483 TI - Effect of dietary versus pharmacological correction of hypertriglyceridemia on red blood cell membrane sodium/lithium countertransport activity. AB - An elevated red blood cell Na/Li countertransport (Na/Li CT) is often associated with high blood pressure and metabolic abnormalities. Recent studies suggested that a reduction in serum TG levels is associated with a decrease in Na/Li CT activity. However, it is still unclear if this phenomenon could be originated from systemic metabolic alterations or from modifications of the membrane dynamic properties. Aim of the present study was to investigate whether dietary or pharmacological TG lowering therapy might have a different effect on Na/Li CT activity and related metabolic parameters. Twenty normotensive hyper-TG patients were recruited from the Lipid outpatient Clinic: they had a baseline Na/Li CT activity significantly higher compared with age- and BMI-matched normolipidemic controls (386+/-33 vs 274+/-39 umol/l RBC/h, p<0.05). The patients were randomly prescribed one of the following two-months treatment: Group 1)-triglyceride lowering diet; Group 2)-lipid lowering drug (Gemfibrozil 600 mg b.i.d.). Na/Li CT and metabolic and anthropometric variables were measured at baseline and after 1 and 2 months of treatment. At the end of intervention, there was in both groups a significant and comparable fall in plasma triglyceride (group 1: -2.61+/-0.73 mmol/l p<0.01; group 2: -4.29+/-1.20 mmol/l p<0.01). In the diet-treated group there were, in addition small but significant reductions in body weight (-3.7+/ 0.8 kg p<0.01), fasting glucose (-0.36+/-0.14 mmol/l p<0.05) and insulin levels ( 2.1+/-0.5 mU/l, p<0.01), while no such changes were observed in the fibrate treated patients. Na/Li CT activity was significantly and comparably reduced at the end of treatment in both groups (group 1: -97+/-28 umol/l cell/h, p<0.01; group 2: -89+/-30 umol/l cell/h, p<0.01). In conclusion, these results indicate that the decrease in Na/Li CT associated with both dietary and drug treatment of hypertriglyceridemia is to be traced to a direct effect of plasma TG concentration on this transport system (probably as a result of modification in the membrane lipid environment) rather than to changes in plasma insulin levels or insulin resistance. PMID- 9199484 TI - Affinities of dopamine analogs for monoamine granular and plasma membrane transporters: implications for PET dopamine studies. AB - Affinities of dopamine (DA) analogs to both granular and plasma membrane uptake transporters were measured in vitro by inhibition of [3H]DA uptake in bovine chromaffin granule ghosts and C6 glial cells transfected with cDNA for the rat presynaptic dopamine transporter, respectively. Five amines were studied: DA, 6 fluorodopamine (6FDA), m-tyramine (MTA), 6-fluoro-m-tyramine (6FMTA), and beta fluoromethylene-m-tyramine (FMMTA). Direct uptake of 18F labeled 6FDA and 6FMTA was also measured in the chromaffin granule system and compared with [3H]DA uptake. Results show that the transporter affinities of 6FDA and MTA were similar to that of DA in both transport systems while affinities of 6FMTA and FMMTA were lower. Furthermore while the direct uptake of DA and FDA in chromaffin granules were essentially identical and significantly reserpine-inhibitable, the direct uptake of 6FMTA was about 15-fold less and only minimally sensitive to reserpine pretreatment. Thus, although vesicular protection and reuptake may influence the turnover of FDA in 6-fluoroDOPA studies, they are unlikely to be important determinants of the kinetics of the slowly clearing components in studies with either 6-fluoro-m-tyrosine (6FMT) or 6-fluoro-beta-fluoro-methylene-m-tyrosine (6FFMMT), the bioprecursors of 6FMTA and 6-fluoro-FMMTA, respectively. These results are consistent with the finding that the longterm component in 6FMT PET studies is 6-fluoro-hydroxyphenylacetic acid (6FHPAC), which can be explained by the lack of vesicular protection of 6FMTA from MAO oxidation. PMID- 9199485 TI - Disruption of the rat mesenteric arterial bed endothelial function by air perfusion. AB - The impact of air perfusion on the endothelial function of the rat mesenteric arterial bed (MAB; perfused with Krebs' bicarbonate plus indomethacin) was compared to that of the NO synthase inhibitor, N(omega)-nitro-L-arginine methyl ester (L-NAME). Air shifted the dose-response curve for the alpha-adrenoceptor agonist, norepinephrine (NE) to the left (ED50%: 2.9+/-0.7 to 0.9+/-0.7 microg, P < 0.05); maximal vasoconstriction did not change. L-NAME produced a similar increase in midrange sensitivity (ED50% 1.4+/-0.7 microg, P < 0.05) and a 20% increase in maximum (152+/-6 to 183+/-7 mmHg, P < 0.05). Electromechanical stimulation with potassium chloride (KCl) was not modified by reserpine. Neither air nor L-NAME modified midrange sensitivity to KCl. L-NAME produced a 17% increase in maximum (91+/-4 to 107+/-5 mmHg, P < 0.05); reserpine abolished the latter effect. Air and L-NAME diminished endothelium-dependent vasodilation elicited by carbachol. Air did not modify endothelium-dependent vasodilation elicited by sodium nitroprusside; this response was potentiated by L-NAME. In summary, air and L-NAME produced similar effects on receptor-dependent activation of the endothelial L-arginine nitric oxide (NO) pathway. Potentiation by L-NAME of the maximal electromechanical response suggests the existence of a tone dependent NO system. Abolition of the latter response by reserpine suggests that this system is of sympathetic origin. PMID- 9199486 TI - One-shot delayed-type hypersensitivity reaction in the mouse liver causes a sustained liver injury to picryl chloride. AB - The developmental characteristics of liver injury induced by a delayed-type hypersensitivity (DTH) mechanism against picryl chloride were examined for 9 consecutive weeks in 3 mouse strains, BALB/c, Kunming and ICR mice. The changes of most biochemical parameters were similar in these three strains, namely, the activities of serum transaminases, lactic dehydrogenase, and prolidase were elevated significantly on day 1, during the first several weeks, and almost throughout the duration, respectively, of liver injury. The content of liver hydroxyproline was also increased after 1-9 weeks of liver injury. In addition, a significant decrease of liver weight, serum alkaline phosphatase and albumin level was observed in BALB/c and Kunming mice. Similar changes in liver histology were also found in the three strains. The hepatocellular necrosis and inflammatory infiltration into the portal area were the predominant features on day 1 and were still distinct during the subsequent several weeks. The mild or moderate hepatocellular degeneration, regeneration and connective tissue hyperplasia were observed after 1 or 3 weeks. A bridging necrosis between portal and portal was observed in several BALB/c and ICR mice, reflecting the possibility of exacerbation of liver injury. These results suggest that the liver injury could be caused and sustained by a one-shot DTH reaction to picryl chloride. The chronicity of the biochemical and histopathological characteristics may be helpful in elucidating the mechanisms of chronic development of liver injury. PMID- 9199487 TI - Time course of induction of oxytocin messenger ribonucleic acid levels in the hypothalamic paraventricular nucleus of ovariectomized rats following gonadal steroid administration. AB - The nonapeptide oxytocin (OT) is important for uterine contractility at parturition, milk ejection during lactation, and the induction of maternal behavior. OT messenger ribonucleic acid (mRNA) levels increase in the paraventricular and supraoptic nuclei (PVN and SON) of late pregnant and lactating rats and are modulated by the steroid milieu that accompanies these states. Specifically, sequential exposure to estradiol (E2) and progesterone (P) followed by P withdrawal 48 hrs prior to sacrifice increases PVN, and to a lesser but significant degree, SON OT mRNA. To better define the time course of induction of OT mRNA levels following P withdrawal, ovariectomized Sprague-Dawley rats were treated with empty or steroid-filled capsules. On day 1, animals received an E2-filled or empty capsule, followed by P-filled or empty capsules on day 3. On day 14, P-filled or empty capsules were removed and animals were sacrificed 24, 36, or 48 hrs later. The hypothalamic PVN were analyzed for OT mRNA by in situ hybridization histochemistry. Significant differences in PVN OT mRNA were found among the groups (P<0.0001, Kruskal-Wallis). Animals in the 48 hr (P=0.007) and 36 hr (P=0.005), but not the 24 hr, steroid-treated groups had significantly increased OT mRNA relative to their respective sham-treated cohorts (Mann-Whitney U test). The relative abundance of PVN OT mRNA differed among the steroid-treated groups (Kruskal-Wallis, P<0.0003), with highest levels at 48 hr. We conclude that increases in PVN OT mRNA occur by 36 hrs, and are highest at 48 hrs, after P withdrawal in the E2-primed rat. Future studies will determine if OT mediated changes in behavior or physiology that surround parturition are related to these changes in OT mRNA. PMID- 9199488 TI - Comparison of the inhibitory effect of isothiourea and mercapto-alkylguanidine derivatives on the alternative pathways of arginine metabolism in macrophages. AB - Novel, non-arginine based compounds have been identified as potent inhibitors of nitric oxide synthase (NOS). Members of the isothiourea and mercapto alkylguanidine classes have generated much interest, as some members of these classes show selectivity towards the inducible isoform of NOS (iNOS), which plays a role in inflammation and shock. Here we compared the effect of a number of these compounds as well as L-arginine based NOS inhibitor reference compounds on macrophage-derived and liver arginase and macrophage iNOS activities. From the non-arginine based NOS inhibitors studied only S-aminoethyl-isothiourea (AETU) caused a slight inhibition of arginase activity. This inhibition was kinetically competitive and due to the rearrangement of AETU to mercapto-ethylguanidine (MEG). The weak inhibitory effect of non-arginine based iNOS inhibitors on arginase activity further supports the view that such compounds may be of practical use for inhibition of NO production in cells simultaneously expressing iNOS and arginase. PMID- 9199489 TI - Effects of fluoxetine on the immunosuppressive response to stress in mice. AB - Mice exposed to a chronic auditory stressor and treated with fluoxetine (5 mg/kg) showed a reduction in stress-induced suppression of thymus and spleen cellularity, and in peripheral T lymphocyte population. The blastogenic response of spleen lymphoid cells and the delayed type hypersensitivity response (DTH) to sheep red blood cells (SRBC) were also assessed and fluoxetine was found to partially reverse the inhibitory effect of stress on both parameters. PMID- 9199490 TI - Bleomycin-induced lung injury is enhanced by interferon-alpha. AB - We have evaluated the effect of intraperitoneal (I.P.) injection of human recombinant interferon-2alpha (IFN-alpha) on Bleomycin-induced pulmonary injury in hamsters. Pulmonary injury was induced by a single intratracheal (I.T.) instillation of Bleomycin (Bleo). Six groups of male Syrian hamsters were treated as follows: 1) I.T. Bleo and daily I.P. injections of low-dose interferon-alpha (2 x 10(4) U), 2) I.T. Bleo and daily I.P. injections of high-dose interferon alpha (10(5) U), 3) I.T. Bleo and I.P. injections of saline, 4) I.T. saline and I.P. low-dose IFN-alpha, 5) I.T. saline and I.P. high-dose IFN-alpha, 6) I.T. saline and I.P. saline. Animals were sacrificed 28 days after I.T. treatment. Lung injury was evaluated histologically and biochemically. Treatment of hamsters with low-dose but not high-dose IFN-alpha significantly augmented the Bleo induced lung injury, as determined by a semiquantitative morphological index. Lung hydroxyproline measurements were highest in Bleo-low-dose-IFN-alpha followed by Bleo-high-dose-IFN-alpha and Bleo-Sal as compared to Sal-Sal and Sal-IFN-alpha controls. These results suggest that IFN-alpha augments Bleo-induced lung injury but that this effect is complex and does not follow a simple-dose-response pattern. PMID- 9199491 TI - Are dipyridamole (sensitive) calcium channels present in esophageal smooth muscle? AB - Calcium (Ca2+) entry from the extra-cellular space into the cytoplasm through voltage-dependent Ca2+ channels, specifically dipyridamole (DHP) sensitive ones (L-type), control a variety of biological processes, including excitation contraction coupling in vascular and GI muscle cells. It has also been proposed that these channels may control esophageal contractility. However, DHP-sensitive Ca2+ channels in esophagus have not been well characterized biochemically. Thus, it is not known if these channels are similar in number or affinity to those in vascular or neural tissues--organs for which clinical use of calcium channel blockers has been successful. Thus, the purpose of this study was to identify and characterize DHP-sensitive calcium channels in esophagus and compare them to vascular, neural, and other GI tissues. METHODS: We carried out in vitro receptor binding assays on lower esophageal muscle homogenates, gastric and intestinal and colonic homogenates, and aortic muscle homogenates from ca; and on brain homogenates from rat. We used a radio-labeled dihydropyridine derivative [3H]nitrendipine, to label these sites and co-administration of unlabeled nimodipine to define specific binding. RESULTS: As expected, ligand binding to L type Ca2+ channels in aortic vascular smooth muscle and brain was readily detectable: brain, Bmax=252 fmol/mg protein, Kd=0.88 nM; aorta, Bmax=326 fmol/mg protein, Kd=0.84 nM. For esophagus (Bmax=97; Kd=0.73) and for other GI tissues, using the same assay conditions, we detected a smaller signal, suggesting that L type Ca2+ channels are present in lower quantities. CONCLUSION: L-type Ca2+ channel are present in esophagus and in other GI muscles, their affinity is similar, but their density is relatively sparse. These findings are consistent with the relatively limited success that has been experienced clinically in the use of calcium channel blockers for treatment of esophageal dysmotility. PMID- 9199492 TI - Calcium- and pH-linked oligomerization of sorcin causing translocation from cytosol to membranes. AB - Sorcin, a cytosolic calcium-binding protein containing a pair of EF-hand motifs, undergoes a Ca2(+)-dependent translocation to the cell membrane. The underlying conformational change is similar at pH 6.0 and 7.5 and consists in an increase in overall hydrophobicity that involves the aromatic residues and in particular the two tryptophan residues which become less exposed to solvent. The concomitant association from dimers to tetramers indicates that the tryptophan residues, which are located between the EF-hand sites, become buried at the dimer-dimer interface. Ca2(+)-bound sorcin displays a striking difference in solubility as a function of pH that has been ascribed to the formation of calcium-stabilized aggregates. PMID- 9199493 TI - Effect of mutations at Cys237 on the activation state and activity of human phenylalanine hydroxylase. AB - Wild-type recombinant human phenylalanine hydroxylase (wt-hPAH) is activated about 1.5-fold by exposure to alkaline pH (pH 8.5-9.0). In order to study whereas this activation might be related to the activation of the rat enzyme by N ethylamaleimide-modification of Cys237 [Gibbs and Benkovic (1991) Biochemistry 30, 67951, mutant proteins of hPAH with Cys237 changed to Ser (S) or Glu (D) have been prepared. The mutant forms have high specific activity at pH 7.0 and high affinity for L-Phe, notably for hPAH-C237D, which shows a 3-fold higher activity than L-Phe-activated wt-hPAH and it is not further activated by pre-incubation with L-Phe. Moreover, the emission maximum of the intrinsic fluorescence of hPAH C237D (lambda(maxem) = 347 nm) resembles that of activated forms of wt-hPAH. However, the activity of this mutant at neutral pH is further activated by exposure to alkaline pH, indicating that activation of wt-hPAH by alkaline pH is not restricted to ionization of Cys237. PMID- 9199494 TI - Bioavailability of rutin and quercetin in rats. AB - Quercetin is a powerful antioxidant which is widely distributed in edible plants, mainly as glycosides such as rutin. It has been reported to be absorbed in mammals, but its metabolism needs further investigation to evaluate its possible physiological effects. We compared the evolution of the absorption of quercetin and rutin in rats fed with supplemented diets. Rutin was absorbed more slowly than quercetin because it must be hydrolysed by the cecal microflora, whereas quercetin was absorbed from the small intestine. Conjugated derivatives of quercetin, and its methylated forms isorhamnetin and tamarixetin, were recovered in plasma from rats receiving the two kinds of experimental diets after the first meal, but after 10 days, no traces of tamarixetin were detected anymore. The rate of elimination of quercetin metabolites seems very low, and high plasma concentrations are easily maintained with a regular supply of quercetin or rutin in the diet. PMID- 9199495 TI - Lipoxygenase inhibitors block CD95 ligand-mediated apoptosis of human malignant glioma cells. AB - CD95 ligand is a cytotoxic cytokine that induces apoptosis. Here we report that CD95-mediated apoptosis of human malignant glioma cells is associated with arachidonic acid (AA) release. Inhibitors of phospholipase A2, phospholipase C or diacylglycerol lipase have minor effects on AA release and fail to modulate apoptosis. Formation of two AA metabolites generated during CD95-dependent apoptosis is attenuated by the lipoxygenase inhibitor, nordihydroguaretic acid (NDGA). NDGA also blocks CD95 ligand-induced apoptosis. This effect is independent of antioxidant properties of NDGA. Lipoxygenase may thus play a critical role in CD95 ligand-induced apoptosis of human malignant glioma cells. PMID- 9199496 TI - The basal subcellular distribution of beta-adrenergic receptor kinase is independent of G-protein beta gamma subunits. AB - beta-Adrenergic receptor kinase (beta ARK-1 or GRK2) is a key regulatory protein involved in the regulation of G-protein-coupled receptors which associates with microsomal and plasma membranes. beta gamma Subunits of G-proteins have been suggested to mediate agonist-dependent membrane translocation of beta ARK, but their possible role in maintaining the complex subcellular distribution of the kinase is not known. In this study we show that lovastatin-mediated inhibition of G gamma subunits isoprenylation in HEK-293 cells stably transfected with beta ARK1 leads to a significant release of G beta subunits to the cytosol without causing changes in total particulate beta ARK or in the association of this kinase to plasma or microsomal membrane fractions. In addition, transient overexpression of mutant forms of G gamma unable to become isoprenylated resulted in a marked sequestration of G beta to the soluble compartment, but caused no rearrangement in the distribution of cotransfected beta ARK. These results indicate that anchoring of beta ARK to cellular membranes under basal conditions is independent of the availability of heterotrimeric G-protein subunits. PMID- 9199497 TI - Hydrolysis of AMPPNP by the motor domain of ncd, a kinesin-related protein. AB - AMPPNP was found to be hydrolyzed by the motor domain of ncd (the product of a Drosophila gene, non-claret disjunctional), a kinesin-related protein. This hydrolysis could be monitored by 31P NMR spectroscopy and by an assay of phosphate, one of the products of the hydrolysis. The rate was approximately 0.00004 s(-1), 1% of the ATP turnover rate. The AMPPNP turnover was not stimulated by microtubules. Kinesin motor domain also turned over AMPPNP but at a somewhat lower rate. Although the turnover was slow, the present finding may present an important caveat, since AMPPNP has been widely used for investigations of kinesin and kinesin-related proteins as a non-hydrolyzable ATP analogue. PMID- 9199498 TI - Overexpression of a nuclear protein tyrosine phosphatase increases cell proliferation. AB - PTP-S2 is a widely expressed nuclear protein tyrosine phosphatase which shows increased expression upon mitogenic stimulation of a variety of cells. In order to elucidate the role of this enzyme in cell division, stable clones of HeLa cells expressing rat PTP-S2 were isolated and their growth properties analysed. Overexpressed PTP-S2 was located in the cell nucleus and there was no significant change in the total tyrosine phosphatase activity of PTP-S2 overexpressing cells. PTP-S2 overexpressing clones, D3 and B5, showed increased rate of cell division and lower serum requirement as compared with control cells. D3 and B5 cells formed larger colonies in soft agar, were not contact inhibited upon confluency, grew in multilayers, and showed altered morphology. These results are consistent with the suggestion that PTP-S2 may be a positive regulator of cell proliferation. PMID- 9199499 TI - The Rhodobacter sphaeroides flagellar motor is a variable-speed rotor. AB - The rotation rate of the unidirectional stop/start motor of Rhodobacter sphaeroides was investigated using computerised motion analysis of tethered cells. The R. sphaeroides motor was found to have a variable rotation rate compared to the virtually constant-speed motor of wild-type and CheR mutant (smooth swimming) Escherichia coli. In addition, the dynamics of the R. sphaeroides motor during stopping was analysed with no consistent correlation behaviour. The motor could go from full rotation to stop, or stop to full rotation within one video frame, i.e. 0.02 s, but it could also slow down into a stop or restart slowly, taking up to 0.25 s. The R. sphaeroides motor under chemokinetic stimulation was also analysed and was found to show increased torque generation and reduced variation in rotation rate. PMID- 9199500 TI - The Wilms tumor suppressor gene WT1 induces G1 arrest and apoptosis in myeloblastic leukemia M1 cells. AB - WT1 was isolated as a tumor suppressor gene of Wilms tumor. However, high expression of WT1 correlates with poor prognosis in acute leukemia. In addition suppression of WT1 expression by WT1 anti-sense oligonucleotide inhibits proliferation of leukemia cells, suggesting that WT1 is important for their proliferation. To further elucidate the biological significance of WT1 in leukemic cell growth, we overexpressed exogenous WT1 in murine M1 myeloblastic leukemia cells using the isopropyl-beta-D-thiogalactoside (IPTG)-controlled expression system. We found that induction of one splicing variant of WT1 [WT1 17AA(+)-KTS(-)] in M1 cells induces cell cycle arrest and apoptotic cell death. These results suggest that the role of WT1 is different depending on the type of leukemia cell in which it is expressed. PMID- 9199501 TI - Nitric oxide protects endothelial cells from tumor necrosis factor-alpha-mediated cytotoxicity: possible involvement of cyclic GMP. AB - In cultured endothelial cells, incubation with TNF-alpha (50 ng/ml) for 48 h markedly reduced viability of endothelial cells. A 6 h preincubation with Sper/NO (0.03--1 microM) protected endothelial cells in a concentration-dependent manner and increased viability by 63% of control. The NO scavenger PTIO (30 microM) completely abolished cytoprotection by Sper/NO. A cytoprotective effect comparable to Sper/NO was observed when preincubating the cells with 8-bromo cyclic GMP (1-10 microM). Moreover, no protection by Sper/NO occurred in the presence of ODQ (0.1 microM), a selective inhibitor of soluble guanylyl cyclase. Our results demonstrate that NO produces a long-term endothelial protection against cellular injury by TNF-alpha, presumably via a cyclic GMP-dependent pathway. PMID- 9199502 TI - A single serine residue confers tetrodotoxin insensitivity on the rat sensory neuron-specific sodium channel SNS. AB - Sensory neurons express a sodium channel (SNS) that is highly resistant to block by tetrodotoxin (IC50 = 60 microM). SNS is 65% homologous to the cardiac sodium channel, in which a single hydrophilic residue in the SS2 segment is critical for tetrodotoxin resistance. By site-directed mutagenesis, we have substituted phenylalanine for serine at the equivalent position in SNS: this mutated (S356F) SNS channel is functionally similar to wild-type SNS when expressed in Xenopus oocytes, but is potently blocked by tetrodotoxin and saxitoxin with IC50s of 2.8 nM and 8.2 nM, respectively. These data provide clues to the rational design of selective blockers of SNS with potential as analgesic drugs. PMID- 9199503 TI - The neuronal nitric oxide synthase PDZ motif binds to -G(D,E)XV* carboxyterminal sequences. AB - PDZ motifs are small protein-protein interaction modules that are thought to play a role in the clustering of submembranous signalling molecules. The specificity and functional consequences of their associative actions is still largely unknown. Using two-hybrid methodology we here demonstrate that the PDZ motif of neuronal nitric oxide synthase (nNOS) can mediate the binding to several other proteins in brain. Peptide library screening showed that proteins bearing a carboxy-terminal G(D,E)XV* sequence are preferred targets for the nNOS amino terminal PDZ motif. Potential nNOS targets include a melanoma-associated antigen, cyclophilins and the alpha1C-adrenergic receptor. PMID- 9199504 TI - Phosphorylation of microtubule-associated protein tau by stress-activated protein kinases. AB - The paired helical filament, which comprises the major fibrous element of the neurofibrillary lesions of Alzheimer's disease, is composed of hyperphosphorylated microtubule-associated protein tau. Many of the hyperphosphorylated sites in tau are serine/threonine-prolines. Here we show that the stress-activated protein (SAP) kinases SAPK1gamma (also called JNK1), SAPK2a (also called p38, RK, CSBPs, Mpk2 and Mxi2), SAPK2b (also called p38beta), SAPK3 (also called ERK6 and p38gamma) and SAPK4 phosphorylate tau at many serine/threonine-prolines, as assessed by the generation of the epitopes of phosphorylation-dependent anti-tau antibodies. Based on initial rates of phosphorylation, tau was found to be a good substrate for SAPK4 and SAPK3, a reasonable substrate for SAPK2b and a relatively poor substrate for SAPK2a and SAPK1gamma. Phosphorylation of tau by SAPK3 and SAPK4 resulted in a marked reduction in its ability to promote microtubule assembly. These findings double the number of candidate protein kinases for the hyperphosphorylation of tau in Alzheimer's disease and other neurodegenerative disorders. PMID- 9199505 TI - Stimulatory effects of parathyroid hormone and 1,25-dihydroxyvitamin D3 on insulin-like growth factor-binding protein-5 mRNA expression in osteoblastic UMR 106 cells: the difference between transient and continuous treatments. AB - The transient treatment with parathyroid hormone (PTH) for 12 h, followed by its removal for 36 h, stimulated insulin-like growth factor-binding protein (IGFBP)-5 mRNA expression more strongly than the continuous treatment for 48 h in osteoblastic UMR-106 cells. The transient but not continuous treatment with A23187 also stimulated it. In contrast, 1,25-dihydroxyvitamin D3 stimulated it, irrespective of the treatment design. IGFBP-5 stimulated type-1 procollagen mRNA expression. The present study first indicated that the transient treatment with PTH more effectively stimulated IGFBP-5 mRNA expression than its continuous treatment partly via an increase in intracellular calcium and suggested that IGFBP-5 might be involved in the anabolic action of PTH in bone. PMID- 9199506 TI - Xanthones as antimalarial agents; studies of a possible mode of action. AB - We recently demonstrated that 2,3,4,5,6-pentahydroxyxanthone (X5) inhibits the in vitro growth of both chloroquine-sensitive and multidrug-resistant strains of P. falciparum. To study the molecular basis of its antimalarial action, we tested X5 and selected hydroxyxanthone analogs as inhibitors of in vitro heme polymerization in a low ionic strength phosphate solution at mildly acidic pH. We found that addition of 1 Eq. of X5 resulted in complete inhibition of polymerization in this system whereas addition of up to 40 Eqs. of standard antimalarial compounds (chloroquine, primaquine, quinacrine, artemisinin and methylene blue) had no such effect although these compounds did co-precipitate with heme. The antimalarial potency of the hydroxyxanthones correlated well with their ability to inhibit in vitro heme polymerization in our assay, suggesting that these compounds exert their antimalarial action by preventing hemozoin formation. Based on the observed structure-activity relationships, we propose a model displaying possible interactions between hydroxyxanthones and heme. PMID- 9199507 TI - Human Supt5h protein, a putative modulator of chromatin structure, is reversibly phosphorylated in mitosis. AB - The Saccharomyces cerevisiae proteins Spt4p, Spt5p and Spt6p are involved in transcriptional repression by modulating the structure of chromatin. From HeLa cells we have purified a human homologue of Spt5p, Supt5hp, and show here that the protein is reversibly phosphorylated in mitosis. The cloned cDNA predicts a protein of 1087 residues with 31% identity to yeast Spt5p. It includes an acidic N-terminus, a putative nuclear localization signal and a C-terminal region containing two different repeated motifs. One of them, with the consensus sequence P-T/S-P-S-P-Q/A-S/G-Y, is similar to the C-terminal domain in the largest subunit of RNA polymerase II. PMID- 9199508 TI - Repression of interleukin-6 gene expression by 17 beta-estradiol: inhibition of the DNA-binding activity of the transcription factors NF-IL6 and NF-kappa B by the estrogen receptor. AB - The cytokine interleukin-6 (IL-6), a key mediator of immune and acute phase responses of the liver, has also been implicated in uterine functions. Estrogens are potent repressors of IL-6 production by uterine stromal cells. In the endometrial adenocarcinoma cell line Ishikawa, phorbol ester-induced activation of the IL-6 promoter was inhibited to basal levels by 17 beta-estradiol (E2) in a wild-type receptor-dependent fashion. Although tamoxifen has been shown to have estrogenic effects on the endometrium, it did not inhibit induction of the IL-6 promoter. We previously showed that inhibition of IL-6 gene expression by E2 does not involve high-affinity binding of the estrogen receptor (ER) to IL-6 DNA. We now report that the ER can directly interact with the transcription factors NF IL6 and NF-kappa B and can inhibit their ability to bind DNA which might be the molecular basis for repression of IL-6 gene expression by estrogens. PMID- 9199509 TI - Analysis of phosphofructokinase subunits and isozymes in ascites tumor cells and its original tissue, murine mammary gland. AB - Phosphofructokinase (PFK) subunits and isozymes have been examined in ascites tumor cells and murine mammary gland, the tissue from where this tumor originated. Ascites tumor was found to contain predominantly the C-type subunit, whereas the L-type subunit was more abundant in mammary gland. An altered M-type subunit of lower electrophoretic mobility was found in both cell types and no M4 homotetramers were detected in either of them. Characteristic regulatory properties of ascites tumor PFK, i.e. insensitivity to fructose-1,6-P2 activation and inhibition by P-enolpyruvate, were also observed in the mammary gland isozyme. The nature of these properties and the contribution of the distinct subunit types to fructose-1,6-P2 activation are discussed. PMID- 9199510 TI - Liposome-mediated peptide loading of MHC-DR molecules in vivo. AB - Amino acid residues 3-15 of mycobacterial HSP60 define a dominant T-cell epitope for HLA-DR3+ve humans and Mamu-DR3+ve rhesus monkeys. Our results show that Mamu DR3 molecules on PBMC can be efficiently loaded in vivo with the above-mentioned peptides when they are intravenously injected encapsulated in liposomes, but not in the free form. Mamu-DR3 loading is abolished by encapsulation of a nonstimulatory peptide. These results have implications for the delivery of therapeutic peptides in vivo. PMID- 9199511 TI - The transcription of NAM7/UPF1 is enhanced in the absence of Cyp1p/Hap1p concomitant with the appearance of an ISF1-NAM7 cotranscript in Saccharomyces cerevisiae. AB - The two adjacent nuclear genes ISF1 and NAM7 cooperatively participate in mitochondrial functions. It is well known that Cyp1p(Hap1p) activates a number of genes involved in these same functions. We show in this paper that Cyp1p influences the transcriptional regulation of NAM7. In addition, a significant amount of ISF1-NAM7 cotranscript is observed in a cyp1 mutant context. An extensive analysis of the intergenic region which separates the two genes revealed 5' starts of the NAM7 transcripts, additional to those previously mapped. These new 5' starts overlap the 3' ends of ISF1. We propose that NAM7 is under the control of a negative Cyp1p-dependent regulator and that its absence favours a transcriptional read-through which results in the ISF1-NAM7 cotranscript we have identified. PMID- 9199512 TI - The hydrophobic probe 4,4'-bis(1-anilino-8-naphthalene sulfonic acid) is specifically photoincorporated into the N-terminal domain of alpha B-crystallin. AB - Photoincorporation of the fluorescent probe 4,4'-bis(1-anilino-8-naphthalene sulfonic acid) (bis-ANS) can be used to locate solvent-exposed hydrophobic regions in proteins. We show that bis-ANS is specifically incorporated into the putative N-terminal domain of alpha B-crystallin. This incorporation diminishes the chaperone-like activity of alpha B-crystallin, suggesting that hydrophobic surfaces in the N-terminal domain are involved in the binding of unfolding proteins. PMID- 9199513 TI - Affinity for alpha-tocopherol transfer protein as a determinant of the biological activities of vitamin E analogs. AB - alpha-Tocopherol transfer protein (alphaTTP), a product of the gene which causes familial isolated vitamin E deficiency, plays an important role in determining the plasma vitamin E level. We examined the structural characteristics of vitamin E analogs required for recognition by alphaTTP. Ligand specificity was assessed by evaluating the competition of non-labeled vitamin E analogs and alpha [3H]tocopherol for transfer between membranes in vitro. Relative affinities (RRR alpha-tocopherol = 100%) calculated from the degree of competition were as follows: beta-tocopherol, 38%; gamma-tocopherol, 9%; delta-tocopherol, 2%; alpha tocopherol acetate, 2%; alpha-tocopherol quinone, 2%; SRR-alpha-tocopherol, 11%; alpha-tocotrienol, 12%; trolox, 9%. Interestingly, there was a linear relationship between the relative affinity and the known biological activity obtained from the rat resorption-gestation assay. From these observations, we conclude that the affinity of vitamin E analogs for alphaTTP is one of the critical determinants of their biological activity. PMID- 9199515 TI - Plasticity of responses to off-vertical axis rotation. AB - This study assessed whether a change in the magnitude of the angular vestibulo ocular reflex (VOR) influences the magnitude of the linear VOR, thereby suggesting a common gain element for these reflexes. The responses to linear acceleration using yaw off-vertical axis rotation (OVAR) at 30 degrees tilt were recorded before and after an adaptation protocol designed to increase the angular VOR gain. Subjects included eight asymptomatic healthy young individuals. Eye movements, recorded with electro-oculography, were analyzed to yield gain of the horizontal angular VOR and the magnitude of the modulation and bias components of the response to OVAR. Results indicated that there was no consistent influence of angular VOR gain on the eye movement response to OVAR. Since previous studies have shown that responses to earth horizontal axis rotation are influenced consistently by angular VOR gain, our study suggests that high intensity otolithic stimulation is required to observe changes in the linear VOR following modification of the angular VOR. PMID- 9199514 TI - The alpha-amylase from the yellow meal worm: complete primary structure, crystallization and preliminary X-ray analysis. AB - The alpha-amylase from Tenebrio molitor larvae (TMA) was purified from a crude larval extract. After removal of the N-terminal pyroglutamate residue and identification of the following 17 residues by Edman sequencing, the cDNA of mature TMA was cloned from larval mRNA. The encoded enzyme consists of 471 amino acid residues and has 57-79% sequence identity to other insect alpha-amylases and also shows high homology to the mammalian enzymes. TMA was crystallized in form of well-ordered orthorhombic crystals of space group P2(1)2(1)2(1) diffracting beyond 1.6 A resolution with unit cell dimensions of a = 51.24 A, b = 93.46 A, c = 96.95 A. TMA may serve as model system for the future analysis of interactions between insect alpha-amylase and proteinaceous plant inhibitors on the molecular level. PMID- 9199516 TI - Electroneurography in the late period of Bell's palsy. AB - Our purpose is to investigate the electrophysiological characteristics of Bell's palsy and to obtain clues for estimating prognosis in the late period by using electroneurography. Thirty-three patients were followed by electroneurography over a period of 12 months. They were classified according to House-Brackman system. At the end of the follow-up, 100% of grade II-III patients, and 61% of grade IV patients recovered completely. Thirty per cent of grade IV patients recovered as grade II, and one grade IV (8%) and one grade V (100%) patient had bad prognoses (grade IV). There were significant differences between each group in the time course between the first and third months of onset. We concluded that the amount of non-degenerated synchronous fibres can allow us to estimate prognosis of Bell's palsy, especially between the first and third month of onset, if we make serial tests. PMID- 9199517 TI - Possible involvement of nitric oxide in the sensorineural hearing loss of bacterial meningitis. AB - Microperfusion of scala tympani with the NO donors, sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine (SNAP), produced marked depression of the compound action potential (CAP) and cochlear microphonic (CM) together with severe and widespread morphological damage to hair cells and supporting cells in the organ of Corti. In addition, direct perfusion of N-methyl-D-aspartate (NMDA) into scala tympani, which probably induces excess stimulation of NMDA receptors within the cochlea and which is known to lead to the release of NO, was found to elicit similar electrophysiological and structural lesions in the cochlea. Pre perfusion of scala tympani with L-methyl arginine (L-MA), which inhibits the release of NO, or superoxide dismutase (SOD), an O2-scavenger, conferred marked protection upon the cochlea from the lesions caused by NO donors. These observations indicate that enhanced NO production is likely to be an important factor responsible for pathological insult of the cochlea. The possibility is discussed that this factor is involved in the chain of events leading to hearing loss caused by bacterial meningitis. Such hearing loss is a major sequela of bacterial meningitis in children. PMID- 9199518 TI - Abnormal audiograms and elevated acoustic reflex thresholds in obligate carriers of autosomal recessive non-syndromic hearing loss. AB - Pure-tone audiograms and acoustic reflex thresholds were obtained in 24 presumed obligate carriers of autosomal recessive non-syndromic hearing loss and 30 sex and age appropriate control subjects, with a view to evaluating the prevalence of abnormalities on these tests in the two groups, and a possible link between the findings on the two tests, which may help to localize threshold deficits and/or abnormal configurations to different sections of the reflex arc. Six (25%) of the carriers and one control subject had abnormal audiograms, inferred to be of genetic aetiology through careful exclusion of environmental risk factors. Four additional carriers had acoustic reflex threshold abnormalities. None of the carriers had an abnormality on both tests. The audiometric configurations and acoustic reflex patterns of abnormality were diverse, and may be a reflection of the genetic heterogeneity in ARNSHL. PMID- 9199519 TI - Latency of contralateral sound-evoked auditory efferent suppression of otoacoustic emissions. AB - The suppression of transiently evoked otoacoustic emissions by contralateral sound stimulation is thought to occur as a result of the action of the efferent pathway from the superior olivary complex to the cochlea via the medial olivo cochlear neurons. The purpose of this study was to determine the time taken for this pathway to activate the suppressive mechanism in response to contralateral sound in normal human subjects. The time for onset of suppression was found to be between 7 and 20 ms. PMID- 9199520 TI - Electrophysiological effects of multiple instillation of Haemophilus influenzae type b endotoxin on the inner ear. AB - An analysis of auditory brainstem response (ABR) thresholds and ABR-based frequency tuning curves was performed in 15 Sprague-Dawley rats exposed to Haemophilus influenzae type b endotoxin; 5 microg/50 microl toxin was instilled every second day, altogether five times, into the middle ear cavity through a small perforation in the tympanic membrane. ABR was measured 48 h after the second application and 24 h, 48 h, 5 days and 10 days after the fifth instillation. Five applications of toxin had no statistical effect on ABR thresholds and no changes in TC configuration were observed. It is concluded that Haemophilus influenzae type b endotoxin, instilled repeatedly through the tympanic membrane into the middle ear, does not affect cochlear electrophysiology. PMID- 9199521 TI - Localization of endothelin-1 and endothelin-3 in the cochlea. AB - The distribution of endothelin-1 (ET-1) and endothelin-3 (ET-3) was studied by indirect immunostaining of decalcified guinea pig and rat cochleae. No species differences were observed. Perikarya and processes of spiral ganglion cells were highly reactive for both ET-1 and ET-3. The epithelial lining of the cochlear duct stained for ET-1 and ET-3, but reactivity for ET-1 was higher in the lining cells of the inner sulcus, Claudius', and Hensen's cells, while the tympanic covering layer of the basilar membrane stained stronger for ET-3 compared to ET 1. In the stria vascularis, all cell types stained for ET-3, while marginal cells were more reactive for ET-1. Spiral ligament fibroblasts were reactive for ET-1, but not for ET-3. Connective tissue cells of the spiral limbus stained for both endothelins. The region of synapses on outer hair cells reacted for ET-1 and ET-3 but sensory cells remained unstained. Endothelins are discussed to act as modulatory peptides, possibly interfering with nitric oxide, prostaglandins, and atrial natriuretic peptide in the lateral cochlear wall (lateral cochlear wall, i.e. stria vascularis and spiral ligament). The occurrence of endothelins in cochlear neurons suggest their potential role as neurotransmitters. PMID- 9199522 TI - Acute study on the neuronal excitability of the cochlear nuclei of the guinea pig following electrical stimulation. AB - To help deaf patients who cannot benefit from the cochlear implant due to interruption of the auditory nerve, a central auditory prosthesis has been developed to directly stimulate the cochlear nucleus in the brainstem. The electrode array lies on the surface of the cochlear nucleus and is designed to stimulate at 250 pulses/sec. To examine the safety of this prosthesis, guinea pig cochlear nuclei were stimulated acutely with bipolar surface electrodes using charge-balanced biphasic current pulses at rates of 250, 500 or 1,000 pulses/s and charge intensities of 1.8, 2.8, 3.5 or 7.1 microC/phase cm(-2). The electrically evoked auditory brainstem response (EABR) was used to monitor neuronal excitability of the cochlear nuclei following this acute electrical stimulation. Respiration rate and body temperature were also monitored during the experiment. The amplitudes and latencies of the EABR waves were measured and compared among the before, during and after stimulation periods. The results showed that respiration rate and body temperature remained within normal limits for the duration of the acute stimulation. During and after electrical stimulation, no change was found in the EABR waveform, dynamic ranges and threshold with up to 6 h direct continuous stimulation of the cochlear nucleus. There was no significant change in the amplitudes and latencies of the EABR waves after stimulation. However, a slight temporary reduction in the amplitude of the EABR waves was observed at 30-60 min during the course of acute stimulation using the highest charge density (7.1 microC/phase cm(-2)). This reduction showed a stronger correlation with the stimulus current, charge/phase and charge density than threshold. The present findings suggest that acute bipolar electrical stimulation with surface electrodes at rates up to 1,000 pulses/s and charge density up to 7.1 microC/phase cm(-2) is safe for neuronal excitability of the cochlear nucleus in guinea pig. PMID- 9199523 TI - Effect of streptomycin intoxication on vestibular nerve regeneration and posture recovery. AB - The action of streptomycin sulfate (SM) on the regenerative process of the vestibular nerve and posture recovery was studied, using bull frogs. The vestibular nerve was sectioned in various conditions with intact endorgan or with SM intoxication. When the nerve was sectioned with the hair cells left intact, the nerve regenerated well and body balance recovered to normal. However, when neural regeneration was blocked, recovery was incomplete. SM intoxication resulted in various degrees of hair cell damage. Degree of posture recovery correlated well with the number of hair cells. When the nerve was sectioned after damaging the hair cell, the nerve failed to regenerate and posture recovery was incomplete. These results suggest that the degree of posture recovery depends on hair cell function and neural regeneration. Furthermore, neural regeneration is strongly influenced by hair cell function. PMID- 9199524 TI - Sequelae of secretory otitis media: changes in middle ear biomechanics. AB - A new method previously introduced investigating the pressure volume relationship of the middle ear system describes dynamic mechanical properties of the system: the variables measured are hysteresis, compliance, and P(ec0) expressing the zero position of the tympanic membrane. The present study investigates the mechanical properties in 69 adolescents treated with ventilation tubes during childhood due to secretory otitis media. The tympanic membranes displayed various degrees of atrophy, sclerosis, and retraction of the pars flaccida. Atrophy was quantitatively related to decreasing hysteresis and increasing compliance, while myringosclerosis showed opposite effects. P(ec0) was significantly lower for the group of former secretory otitis media than for normals (p < 0.001). This reflects a retraction pattern of the tympanic membrane, which may be explained by a low opening pressure of the eustachian tube or previous pressure load of the drum. Signs of retraction were not found by tympanometry. Treatment with ventilation tubes was associated with a dramatic increase of tympanic membrane pathology (66%) compared to untreated ears (12%), as assessed by otomicroscopy (p < 0.001). However, these changes specific to treatment were not found in the corresponding mechanical variables of the middle ear system, as the effects of combined atrophy and myringosclerosis tend to counterbalance. PMID- 9199525 TI - Effect of middle ear pressure change on middle ear mechanics. AB - The effect of graded variations in middle ear pressure on ossicular vibration was measured in 15 normal human temporal bone specimens. The displacement amplitude of the umbo and stapes head was measured at 16 frequencies between 0.2 kHz and 3.5 kHz at a constant sound pressure of 134 dB SPL at the tympanic membrane (TM) using a non-contacting video measuring system. Both negative and positive pressures decreased umbo and stapes vibration at low frequencies and slightly increased the vibration at higher frequencies. The effects were greater for negative pressure than for positive pressure. The change in stapes vibration was less than that of the umbo at low frequencies, but increased at higher frequencies. In some temporal bones, a small positive pressure produced improvement in stapes vibration at all frequencies. These effects were thought to be primarily due to an increased stiffness of the TM and a damping of ossicular vibration, due to stretching of the ossicular suspensory ligaments and the annular ligament of the footplate. PMID- 9199526 TI - The tympanic membrane and middle ear mucosa during non-typeable Haemophilus influenzae and Haemophilus influenzae type b acute otitis media: a study in the rat. AB - Non-typeable Haemophilus influenzae (NTHi) and encapsulated Haemophilus influenzae type b (Hib) were inoculated into the middle ears of Sprague-Dawley rats. Tympanic membrane (TM) status was assessed otomicroscopically and specimens from various middle ear areas were prepared for light microscopy at various times during the acute phase and up to 6 months after inoculation. Irrespective of bacteria strain, acute otitis media (AOM) was present in all ears 4 days after inoculation. The Hib-infected ears showed initially a severe course of AOM, but all were otomicroscopically resolved by day 12, at which time a few NTHi inoculated ears still exhibited middle ear effusion. The TMs infected with Hib had normalized without scar formation, whereas NTHi induced a persistent thickening of the TMs in half of all cases. The middle ear mucosa of NTHi infected ears initially showed vigorous activity among the goblet cells, but the mucosa normalized after the acute phase. Hib, by contrast, induced prominent changes in the middle ear mucosa. Initially, no goblet cell granules or ciliated cells could be observed in the mucosa. Later on, the epithelium contained large, active goblet cells. Glands appeared beneath the mucosa which persisted as streaks of epithelial cells throughout the study period. The findings show that NTHi and Hib both induce AOM but with differing clinical courses, and affect different targets in the middle ear. PMID- 9199528 TI - Possible value of nasal smear in acute maxillary sinusitis: an experimental study. AB - The aim of this study is to determine the role of nasal smear in evaluating diagnosis and response to the treatment by the patient of acute maxillary sinusitis. We compared nasal smear and histopathological findings obtained from rabbits experimentally induced acute maxillary sinusitis. The animals were divided into two groups; one with blocked ostium and treated with antibiotic and the other applied natural ostiotomy, during a 4-week period. There was no statistically significant difference between the two groups in respect of recovery period. This conclusion was derived from nasal smear and biopsy findings. It was observed that nasal smear and biopsy findings were consistent with each other and with clinical findings. The results of this study revealed that nasal smear may be used in the diagnosis and treatment follow-up of acute maxillary sinusitis. PMID- 9199527 TI - Dexamethasone inhibits mucous glycoprotein secretion via a phospholipase A2 dependent mechanism in cultured chinchilla middle ear epithelial cells. AB - Inhibition or attenuation of mucous hypersecretion in middle ear epithelium is a key step toward resolution of mucoid otitis media. Mucous hypersecretion induced by platelet-activating factor (PAF) in cultured Chinchilla middle ear epithelial cells is dependent on arachidonic acid metabolites via PAF receptors, suggestive of the role of phospholipase A2 (PLA2) in mucous glycoprotein (MGP) secretion. In this study, dexamethasone added to cultured Chinchilla middle ear epithelial cells inhibited baseline and PAF-induced MGP secretion in a concentration dependent manner. A definite time lag (16 h) was observed between administration of dexamethasone and MGP inhibition. This inhibition was reversed by the addition of exogenous PLA2 (the rate-limiting enzyme of arachidonic acid metabolism) and actinomycin D (an inhibitor of mRNA synthesis). This suggests that dexamethasone inhibits baseline and PAF-induced MGP secretion via a PLA2-dependent mechanism. PMID- 9199529 TI - Reduced sialylation of glycoproteins in nasal glands of patients with chronic sinusitis. AB - This study was conducted to investigate the sialylations of glycoproteins in the nasal glands of patients with chronic sinusitis. Sialic acids were detected using lectin histochemistry, and the mRNA of sialyltransferase was evaluated by in situ hybridization histochemistry. Sambucus nigra agglutinin (SNA), which recognizes terminal sialic acids, strongly stained the glandular mucous cells of normal subjects, but not those of patients with chronic sinusitis. In situ hybridization histochemistry showed that the expression of alpha2,6 sialyltransferase mRNA was decreased in the secretory cells of patients with chronic sinusitis. Our present results suggest that a reduction in sialyltransferase activity at the mRNA level in the nasal glands may lead to the persistence of chronic sinusitis. PMID- 9199530 TI - Nasal airway dimensions in term neonates measured by continuous wide-band noise acoustic rhinometry. AB - The nasal airways of 94 healthy term infants (37-42 weeks gestational age) were examined by continuous wide-band noise acoustic rhinometry under standardized conditions on the second or third day postpartum. Validation of the flexible infant probe on tubular plastic models with dimensions similar to the nasal cavity of newborn infants showed a correlation coefficient of 0.98. The mean and standard deviation (SD) of the total minimal cross-sectional area (TMCA), the distance from the nostril to the MCA (DMCA) and the total volumes between the nostril and 45 mm into the nasal cavity (TVOL45) were 0.20 +/- 0.05 cm2, 0.76 +/- 0.29 cm and 2.14 +/- 0.39 cm3 respectively. In general, both anthropometric and rhinometric mean values were higher in males (n = 52) compared to females (n = 42), and the difference was statistically significant for TMCA and head circumference. We conclude that the technical properties, small size and flexible tube of the miniprobe make it uniquely suited for objective assessment of the nasal airways in infants and small children. The RHIN 2000/2100 miniprobe (S.R. Electronics Aps, Lynge, Denmark) is the first infant probe commercially available, making standardization and comparison of results easier. PMID- 9199531 TI - Metachromatic cells and eosinophils in the nasal mucosa and N,N dimethylbenzylamine exposure. AB - Six healthy non-atopic male volunteers participated in a dose-response study of N,N-dimethylbenzylamine (DMBA), which is a reactive chemical used in epoxy systems. The effects on the nasal mucosa after inhalation of 0, 20, 45, 80 and 120 microg/m3 were studied by means of symptom recordings, acoustic rhinometry, nasal lavages and nasal cytology processed for light microscopy of metachromatic cells (MC) and eosinophils (EOS). Although only minor symptoms were provoked, the numbers of MC and Eos tended to increase in a dose-response fashion after inhalation of the chemical. No signs of degranulation of the cells were found, as the levels of tryptase and eosinophil cationic protein in the nasal lavages remained low at all DMBA exposure levels. We therefore conclude that a reactive chemical such as DMBA can influence MC and Eos in the nasal mucosa even at low dose levels without causing significant clinical symptoms. PMID- 9199532 TI - Electron spin resonance spin trapping assay and immunohistochemical localization of superoxide dismutases in the rat nasal mucosa. AB - The immunohistochemical method and electron spin resonance (ESR) spin trapping assay were employed to detect the localization and biochemical activity of superoxide dismutases (SODs) in the rat nasal mucosa. Manganese SOD and copper zinc SOD were immunohistochemically illustrated to be richly expressed in the epithelial cells and the subepithelial glands of nasal mucosa. The olfactory vesicles also showed positive immunostaining for manganese SOD and copper-zinc SOD. ESR spin trapping assay revealed that SOD activity in the mucosa of olfactory areas was significantly higher than in the mucosa of respiratory areas; however, the ratio of SOD activity in the mitochondrial fraction to SOD activity in the cytosolic fraction was similar, approximating 17:83 in the mucosa of both the olfactory and respiratory areas. The predominant localization of SODs in epithelial cells of nasal mucosa suggests the importance of mucosal epithelium in protecting nasal mucosa against cytotoxic superoxide (O2-) radicals. Epithelial goblet cells and the connective tissue of lamina propria, which showed no positive immunostaining for SODs, are considered to be vulnerable to oxidative insults implicated in the generation of O2- radicals. The higher SODs activity in the mucosa of olfactory areas implies that there is a different requirement of SOD in mucosa of the respiratory and olfactory areas on scavenging microenvironmental O2- radicals. PMID- 9199533 TI - Peptidergic innervation in human von Ebner's glands: an immunohistochemical study. AB - The occurrence and distribution of several neuropeptides were studied in human von Ebner's glands. Immunoreactivity for substance P, calcitonin gene-related peptide, vasoactive intestinal polypeptide (VIP), neuropeptide Y (NPY), galanin, and somatostatin was found in the nerve fibers around the acini, ducts, and blood vessels. VIP-immunoreactive varicose fibers were numerous compared with the other five neuropeptides. Most NPY fibers were associated with the vasculature in the gland. These findings suggest that the neuropeptides may regulate the secretion and vascular tone in human von Ebner's glands. PMID- 9199534 TI - Deterioration of the Provox silicone tracheoesophageal voice prosthesis: microbial aspects and structural changes. AB - Device life of tracheoesophageal voice prostheses is limited due to deterioration of the polymers. A group of 55 postlaryngectomy patients fitted with a Provox voice prosthesis have been studied prospectively during 6 months. Thirty-seven prostheses were replaced due to a dysfunctional valve mechanism. Although colonization with Candida species was highly associated with destruction of the silicone material, other upper respiratory tract commensals, e.g. Staphylococcus aureus, were also demonstrated. Electron microscopy of the contaminated devices showed colonization and disruption of the silicone material by penetrating yeast hyphae. During the study a remarkable increase of intratracheal phonatory pressures was assessed with progressive colonization of the prostheses. PMID- 9199535 TI - Electromyographic activity of strap and cricothyroid muscles in pitch change. AB - The EMG activity of the cricothyroid muscle (CT) and the three extrinsic laryngeal muscles (thyohyoid, TH; sternothyroid, ST, and sternohyoid, SH) were recorded throughout the voice range of one female and one male subject, both untrained singers. The voice range was examined using rising and falling glissandos (production of a sustained sound with progressive and continuous variation of fundamental frequency). Muscle activity was observed at various pitches during the glissandos. The strap muscle activity during the production of glissandos appears to be synergistic. At the lowest frequency, the CT is inactive but strap muscles (TH, ST, SH) are active. As frequency increases, strap muscle activity decreases while the CT controls frequency in the middle of the range. At higher frequencies the strap muscles once again become active. This activity might depend on the vocal vibratory mechanism involved. The role of the strap muscles at high pitches is a widely debated point but it seems that in some way they control the phenomena relevant to the rising pitch. The phasic-type strap muscle activity contrasts with the tonic-type activity of the CT. The CT closely controls the frequency, while the straps are not directly linked to the pitch but rather to the evolution of the frequency of voice production (speaking voice, singing voice, held notes, glissandos, trillo, vibrato, etc.). PMID- 9199537 TI - Occupational exposures and amyotrophic lateral sclerosis. A population-based case control study. AB - This population-based case-control study was conducted in three countries in western Washington State to evaluate associations between workplace exposures and the risk of amyotrophic lateral sclerosis (ALS). Cases (n = 174) were all newly diagnosed with ALS by neurologists during 1990-1994, and controls (n = 348), who were matched according to age (+/-5 years) and sex, were identified via random digit dialing or Medicare enrollment files. Four industrial hygienists blindly assessed detailed lifetime job histories for exposures to metals, solvents, and agricultural chemicals. Case-control comparisons were made for jobs held between 15 years of age and 10 years prior to the cases' dates of diagnosis. After adjustment for age and education, ever exposure to agricultural chemicals was associated with ALS (odds ratio (OR) = 2.0, 95% confidence interval (CI) 1.1 3.5); this association was observed separately in men (OR = 2.4, 95% CI 1.2-4.8) but not in women (OR = 0.9, 95% CI 0.2-3.8). Among men, the odds ratio for low exposure to agricultural chemicals (below the median level for exposed controls) relative to no exposure was 1.5 (95% CI 0.4-5.3), and for high exposure, it was 2.8 (95% CI 1.3-6.1) (p for trend = 0.03). Similar analyses based on the panel's assessment of exposures to metals and solvents showed no associations. These findings suggest an association between ALS and agricultural chemicals in men. PMID- 9199536 TI - Cancer mortality in workers exposed to phenoxy herbicides, chlorophenols, and dioxins. An expanded and updated international cohort study. AB - The authors examined cancer mortality in a historical cohort study of 21,863 male and female workers in 36 cohorts exposed to phenoxy herbicides, chlorophenols, and dioxins in 12 countries. Subjects in this updated and expanded multinational study coordinated by the International Agency for Research on Cancer were followed from 1939 to 1992. Exposure was reconstructed using job records, company exposure questionnaires, and serum and adipose tissue dioxin levels. Among workers exposed to phenoxy herbicides contaminated with 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) or higher chlorinated dioxins, mortality from soft-tissue sarcoma (6 deaths; standardized mortality ratio (SMR) = 2.03, 95% confidence interval (CI) 0.75-4.43) was higher than expected from national mortality rates. Mortality from all malignant neoplasms (710 deaths; SMR = 1.12, 95% CI 1.04-1.21), non-Hodgkin's lymphoma (24 deaths; SMR = 1.39, 95% CI 0.89 2.06), and lung cancer (225 deaths; SMR = 1.12, 95% CI 0.98-1.28) was slightly elevated. Risks for all neoplasms, for sarcomas, and for lymphomas increased with time since first exposure. In workers exposed to phenoxy herbicides with minimal or no contamination by TCDD and higher chlorinated dioxins, mortality from all neoplasms (398 deaths; SMR = 0.96, 95% CI 0.87-1.06), non-Hodgkin's lymphoma (9 deaths; SMR = 1.00), and lung cancer (148 deaths; SMR = 1.03) was similar to that expected, and mortality from soft-tissue sarcoma was slightly elevated (2 deaths; SMR = 1.35). In a Poisson regression analysis, workers exposed to TCDD or higher chlorinated dioxins had an increased risk for all neoplasms (rate ratio = 1.29, 95% CI 0.94-1.76) compared with workers from the same cohort exposed to phenoxy herbicides and chlorophenols but with minimal or no exposure to TCDD and higher chlorinated dioxins. These findings indicate that exposure to herbicides contaminated with TCDD and higher chlorinated dioxins may be associated with a small increase in overall cancer risk and in risk for specific cancers. PMID- 9199538 TI - Fatal occupational injuries in a southern state. AB - Fatal occupational injuries were studied using data from medical examiners' reports in North Carolina for the years 1977-1991. Cases were defined as deaths due to accidents or homicide at the workplace, and populations at risk were estimated from the 1980 and 1990 US Censuses. Mortality rate ratios and proportionate mortality ratios were used as measures of association, and the population attributable risk percentage was used as an indicator of the burden of injury. Standard weights for direct age-adjustment of rates were obtained from the total state workforce. There were 2,524 eligible deaths-83 percent from unintentional traumatic injuries, 14 percent from homicide, and the remainder from other causes. This report focuses on unintentional trauma deaths, which were strongly associated with the wood production, fishing, and transportation industries. Elderly, African-American, and self-employed workers had higher fatality rates than members of other groups. Among male workers, motor vehicle crashes were the principal cause of death on the job, followed by falling objects, machinery, and falls. The industries contributing the largest proportions of these deaths were construction, trucking, agriculture, and logging (population attributable risk percentages were 16.8%, 8.8%, 7.9%, and 6.9%, respectively). The fatality patterns of female workers were different: Numbers of deaths from homicide and unintentional trauma were equal, and 27% of the latter deaths occurred in one catastrophic fire. Decentralized and rural industries were the most hazardous, but many deaths were outside the current jurisdiction of occupational safety and health agencies. These patterns suggest that greater scrutiny of such industries, through both research and intervention, is warranted. PMID- 9199539 TI - Prognostic factors and survival of laryngeal cancer patients from Turin, Italy. A population-based study. AB - Little information is available on the role of risk factors for cancer of the larynx in survival. This study analyzed survival through the end of 1994 for 355 cases of laryngeal cancer diagnosed among residents of Turin, Italy, during 1979 1982. Relative survival at 5 years was 75% in women and 67% in men. The role of clinical and etiologic factors was analyzed in detail among 222 male cases. The role of nodal involvement as a strong predictor of poor survival was confirmed. Patients of low socioeconomic status experienced poorer survival than other patients, as did heavy smokers. Alcohol drinking and diet did not seem to strongly influence survival. Survival in this series of laryngeal cancer cases closely parallels that observed in other case series from Europe. While the results regarding socioeconomic status, tobacco smoking, and alcohol drinking parallel those of the few previous studies available, this investigation did not confirm a role for diet in the survival of laryngeal cancer patients, a finding that was recently seen in another study from northern Italy. PMID- 9199540 TI - Blood pressure and performance on the Mini-Mental State Examination in the very old. Cross-sectional and longitudinal data from the Kungsholmen Project. AB - The authors examined the association of blood pressure with cognitive function as assessed by the Mini-Mental State Examination (MMSE) in a community-based Swedish cohort of 1,736 people aged 75-101 years. Age, sex, education, antihypertensive medication use, heart disease, and stroke were considered as covariates. Multiple linear regression analysis indicated that both systolic and diastolic blood pressure, measured in 1987-1989, were positively and significantly related to baseline MMSE score; baseline systolic pressure was also positively and significantly related to follow-up MMSE score, measured after an average period of 40.5 months among subjects who were not taking antihypertensive medication at baseline. Furthermore, in the nontreated group, multiple logistic regression showed that individuals with a baseline systolic pressure less than 130 mmHg had an odds ratio of 1.88 (p = 0.05) for follow-up cognitive impairment (MMSE score < 24) compared with those whose systolic pressure was 130-159 mmHg. An increased but not statistically significant risk of cognitive impairment was associated with high blood pressure (systolic pressure > or = 180 mmHg or diastolic pressure > or = 95 mmHg) only in persons taking antihypertensive medication at baseline. Subjects with systolic pressure of 160-179 mmHg tended to be at lower risk of cognitive impairment. These results may support the view that a certain blood pressure level, particularly a systolic pressure of at least 130 mmHg, is important to the maintenance of cognitive functioning in the very old. They also suggest that severe hypertension that is not well controlled (systolic pressure > or = 180 mmHg or diastolic pressure > or = 95 mmHg) is still a threat to cognitive function in this age group. However, the use of blood pressure measurements made at a single visit and the relatively short follow-up period should be considered when interpreting these results. PMID- 9199541 TI - Fatty acids in serum cholesteryl esters as quantitative biomarkers of dietary intake in humans. AB - The fatty acid composition of serum cholesteryl esters is used as a qualitative biomarker of fatty acid intake, but quantitative data are scarce. Between 1987 and 1992, the authors fed various fatty acids in four controlled trials to 232 healthy Dutch volunteers and measured the proportion of fatty acids in participants' cholesteryl esters. Each 10% of energy fed as linoleic acid (18:2) raised the proportion of linoleic acid in cholesteryl esters by 9.3 g per 100 g of fatty acids (standard deviation (SD) 3.1). For oleic acid (cis-18:1), this figure was 6.5 g/100 g (SD 1.7); for trans fatty acids (trans-18:1), it was 1.1 (SD 0.5); for stearic acid (18:0), 1.0 (SD 0.4); for palmitic acid (16:0), 1.7 (SD 0.5); for myristic acid (14:0), 2.1 (SD 0.7); and for a mixture of saturated fatty acids (12:0, 14:0, and 16:0), it was 2.2 g/100 g (SD 1.0). The coefficient of variation of the responses was fairly constant, indicating that changes in intake for each of these fatty acids can be monitored with similar precision. These data can be used to estimate the degree of compliance in experimental studies involving exchanges of single dietary fatty acids. Most fatty acids in cholesteryl esters may also be used in observational studies to estimate differences in intake. However, because of multiple simultaneous differences in fatty acid intake between free-living individuals and between populations, such data cannot provide information on absolute intake of fatty acids. PMID- 9199542 TI - Concordance of Medicare data and population-based clinical data on cataract surgery utilization in Olmsted County, Minnesota. AB - The authors assessed concordance of local Medicare health care utilization data on cataract surgery and estimates generated using the databases of the Rochester Epidemiology Project, which capture virtually all medical care received by residents of Olmsted County, Minnesota. The Rochester Project databases identified 1,353 primary cataract extractions performed in Olmsted County between October 1989 and December 1993 among county residents aged > or = 65 years. Medicare data identified 1,148 claims-84.8% of the number of procedures identified by the Rochester Project. Ratios of numbers of encounters (Medicare/Rochester Project) were 189/350 (0.540) for 1992 versus 959/1,003 (0.956) for the other years combined. Changes in Medicare data file transfer procedures may have produced the 1992 data shortfall. Medicare data should periodically be compared with source data to assess concordance. PMID- 9199543 TI - Uncertainty and sensitivity analysis of the basic reproductive rate. Tuberculosis as an example. AB - The basic reproductive rate (R0) is a measure of the severity of an epidemic. On the basis of replicated Latin hypercube sampling, the authors performed an uncertainty and sensitivity analysis of the basic reproductive rate of tuberculosis (TB). The uncertainty analysis allowed for the derivation of a frequency distribution for R0 and the assessment of the relative contribution each of the three components of R0 made when TB epidemics first arose centuries ago. (The three components of R0 are associated with fast, slow, and relapse TB.) R0 estimates indicated the existence of fairly severe epidemics when TB epidemics first arose. The R0 for the susceptible persons who developed TB slowly (R0(slow)) contributed the most to the R0 estimates; however, the relative R0(slow) contribution decreased as the severity of TB epidemics increased. The sensitivity of the magnitude of R0 to the uncertainty in estimating values of each of the input parameters was assessed. These results indicated that five of the nine input parameters, because of their estimation uncertainty, were influential in determining the magnitude of R0. This uncertainty and sensitivity methodology provides results that can aid investigators in understanding the historical epidemiology of TB by quantifying the effect of the transmission processes involved. Additionally, this method can be applied to the R0 of any other infectious disease to estimate the probability of an epidemic outbreak. PMID- 9199544 TI - Validity of methods for model selection, weighting for model uncertainty, and small sample adjustment in capture-recapture estimation. AB - In log-linear capture-recapture approaches to population size, the method of model selection may have a major effect upon the estimate. In addition, the estimate may also be very sensitive if certain cells are null or very sparse, even with the use of multiple sources. The authors evaluated 1) various approaches to the issue of model uncertainty and 2) a small sample correction for three or more sources recently proposed by Hook and Regal. The authors compared the estimates derived using 1) three different information criteria that included Akaike's Information Criterion (AIC) and two alternative formulations of the Bayesian Information Criterion (BIC), one proposed by Draper ("two pi") and one by Schwarz ("not two pi"); 2) two related methods of weighting estimates associated with models; 3) the independent model; and 4) the saturated model, with the known totals in 20 different populations studied by five separate groups of investigators. For each method, we also compared the estimate derived with or without the proposed small sample correction. At least in these data sets, the use of AIC appeared on balance to be preferable. The BIC formulation suggested by Draper appeared slightly preferable to that suggested by Schwarz. Adjustment for model uncertainty appears to improve results slightly. The proposed small sample correction appeared to diminish relative log bias but only when sparse cells were present. Otherwise, its use tended to increase relative log bias. Use of the saturated model (with or without the small sample correction) appears to be optimal if the associated interval is not uselessly large, and if one can plausibly exclude an all-source interaction. All other approaches led to an estimate that was too low by about one standard deviation. PMID- 9199545 TI - Behavioral genetics '97: ASHG statement. Recent developments in human behavioral genetics: past accomplishments and future directions. AB - The field of behavioral genetics has enormous potential to uncover both genetic and environmental influences on normal and deviant behavior. Behavioral-genetic methods are based on a solid foundation of theories and methods that successfully have delineated components of complex traits in plants and animals. New resources are now available to dissect the genetic component of these complex traits. As specific genes are identified, we can begin to explore how these interact with environmental factors in development. How we interpret such findings, how we ask new questions, how we celebrate the knowledge, and how we use or misuse this knowledge are all important considerations. These issues are pervasive in all areas of human research, and they are especially salient in human behavioral genetics. PMID- 9199546 TI - Genetic influences in childhood-onset psychiatric disorders: autism and attention deficit/hyperactivity disorder. PMID- 9199547 TI - Understanding the genetic basis of mood disorders: where do we stand? PMID- 9199549 TI - To fire the train: a second malignant-hyperthermia gene. PMID- 9199550 TI - Searching for gene defects that cause high bone mass. PMID- 9199548 TI - Genetics of narcolepsy and other sleep disorders. PMID- 9199551 TI - The great escape. PMID- 9199552 TI - Malignant-hyperthermia susceptibility is associated with a mutation of the alpha 1-subunit of the human dihydropyridine-sensitive L-type voltage-dependent calcium channel receptor in skeletal muscle. AB - Malignant hyperthermia susceptibility (MHS) is characterized by genetic heterogeneity. However, except for the MHS1 locus, which corresponds to the skeletal muscle ryanodine receptor (RYR1) and for which several mutations have been described, no direct molecular evidence for a mutation in another gene has been reported so far. In this study we show that the CACNL1A3 gene encoding the alpha 1-subunit of the human skeletal muscle dihydropyridine-sensitive L-type voltage-dependent calcium channel (VDCC) represents a new MHS locus and is responsible for the disease in a large French family. Linkage analysis performed with an intragenic polymorphic microsatellite marker of the CACLN1A3 gene generated a two-point LOD score of 4.38 at a recombinant fraction of 0. Sequence analysis of the coding region of the CACLN1A3 gene showed the presence of an Arg His substitution at residue 1086, resulting from the transition of A for G3333, which segregates perfectly with the MHS phenotype in the family. The mutation is localized in a very different part of the alpha 1-subunit of the human skeletal muscle VDCC, compared with previously reported mutations found in patients with hypokalemic periodic paralysis, and these two diseases might be discussed in terms of allelic diseases. This report is the first direct evidence that the skeletal muscle VDCC is involved in MHS, and it suggests a direct interaction between the skeletal muscle VDCC and the ryanodine receptor in the skeletal muscle sarcoplasmic reticulum. PMID- 9199553 TI - Linkage of a gene causing high bone mass to human chromosome 11 (11q12-13) AB - The purpose of this paper is to report the linkage of a genetic locus (designated "HBM") in the human genome to a phenotype of very high spinal bone density, using a single extended pedigree. We measured spinal bone-mineral density, spinal Z(BMD), and collected blood from 22 members of this kindred. DNA was genotyped on an Applied Biosystems model 377 (ABI PRISM Linkage Mapping Sets; Perkin Elmer Applied Biosystems), by use of fluorescence-based marker sets that included 345 markers. Both two-point and multipoint linkage analyses were performed, by use of affected/unaffected and quantitative-trait models. Spinal Z(BMD) for affected individuals (N = 12) of the kindred was 5.54 +/- 1.40; and for unaffected individuals (N = 16) it was 0.41 +/- 0.81. The trait was present in affected individuals 18-86 years of age, suggesting that HBM influences peak bone mass. The only region of linkage was to a series of markers on chromosome 11 (11q12 13). The highest LOD score (5.21) obtained in two-point analysis, when a quantitative-trait model was used, was at D11S987. Multipoint analysis using a quantitative-trait model confirmed the linkage, with a LOD score of 5.74 near marker D11S987. HBM demonstrates the utility of spinal Z(BMD) as a quantitative bone phenotype that can be used for linkage analysis. Osteoporosis pseudoglioma syndrome also has been mapped to this region of chromosome 11. Identification of the causal gene for both traits will be required for determination of whether a single gene with different alleles that determine a wide range of peak bone densities exists in this region. PMID- 9199554 TI - Expression of genes from the human active and inactive X chromosomes. AB - X-chromosome inactivation results in the cis-limited inactivation of many, but not all, of the genes on one of the pair of X chromosomes in mammalian females. In addition to the genes from the pseudoautosomal region, which have long been anticipated to escape inactivation, genes from several other regions of the human X chromosome have now been shown to escape inactivation and to be expressed from both the active and inactive X chromosomes. The growing number of genes escaping inactivation emphasizes the need for a reliable system for assessing the inactivation status of X-linked genes. Since many features of the active or inactive X chromosome, including transcriptional activity, are maintained in rodent/human somatic-cell hybrids, such hybrids have been used to study the inactivation process and to determine the inactivation status of human X-linked genes. In order to assess the fidelity of inactivation status in such hybrids, we have examined the expression of 33 X-linked genes in eight mouse/human somatic cell hybrids that contain either the human active (three hybrids) or inactive X (five hybrids) chromosome. Inactivation of nine of these genes had previously been demonstrated biochemically in human cells, and the expression of these genes only in hybrids retaining an active X, but not in those retaining an inactive X, confirms that expression in hybrids reflects expression in human cells. Although the majority of genes tested showed consistent patterns of expression among the active X hybrids or inactive X hybrids, surprisingly, 5 of the 33 genes showed heterogeneous expression among the hybrids, demonstrating a significantly higher rate of variability than previously reported for other genes in either human somatic cells or mouse/human somatic-cell hybrids. These data suggest that at least some X-linked genes may be under additional levels of epigenetic regulation not previously recognized and that somatic-cell hybrids may provide a useful approach for studying these chromosomal phenomena. PMID- 9199555 TI - Three novel homozygous point mutations and a new polymorphism in the COL17A1 gene: relation to biological and clinical phenotypes of junctional epidermolysis bullosa. AB - Junctional epidermolysis bullosa (JEB) is a clinically and biologically heterogeneous genodermatosis, characterized by trauma-induced blistering and healing without scarring but sometimes with skin atrophy. We investigated three unrelated patients with different JEB phenotypes. Patients 1 and 2 had generalized atrophic benign epidermolysis bullosa (GABEB), with features including skin atrophy and alopecia. Patient 3 had the localisata variant of JEB, with predominantly acral blistering and normal hair. All patients carried novel homozygous point mutations (Q1016X, R1226X, and R1303Q) in the COL17A1 gene encoding collagen XVII, a hemidesmosomal transmembrane component; and, therefore, not only GABEB but also the localisata JEB can be a collagen XVII disorder. The nonsense mutations led to drastically reduced collagen XVII mRNA and protein levels. In contrast, the missense mutation allowed expression of abnormal collagen XVII, and epidermal extracts from that patient contained polypeptides of normal size, as well as larger aggregates. The homozygous nonsense mutations in the COL17A1 gene were consistent with the absence of the collagen from the skin and with the GABEB phenotype, whereas homozygosity for the missense mutation resulted in expression of aberrant collagen XVII and, clinically, in localisata JEB. PMID- 9199556 TI - Characterization of FMR1 promoter elements by in vivo-footprinting analysis. AB - Fragile X syndrome is associated with silencing of the FMR1 gene. We studied the transcriptional regulation, by analysis of the FMR1 promoter region for the presence of in vivo protein/DNA interactions and for cytosine methylation at the single-nucleotide level. Four protein-binding sites were present in the unmethylated promoter of the active FMR1 gene. In the methylated promoter of inactive genes no footprints were detected, and no evidence of active repression was found in the region investigated. We propose that the silencing of FMR1 gene transcription results from a lack of transcription-factor binding. PMID- 9199557 TI - Functional and structural features of a tandem duplication of the human mtDNA promoter region. AB - An approximately 260-bp tandem duplication of the human mtDNA regulatory region has been identified in patients with mitochondrial disorders and in a specific Caucasian haplogroup. The functional significance of this mtDNA duplication was difficult to assess, because it was present at very low levels in human tissues. We have isolated several transmitochondrial cybrid lines harboring this mutation, one of which (clone CA17.1) was essentially homoplasmic for the duplication. Oxidative-phosphorylation function was not impaired in clone CA17.1, suggesting that this mtDNA alteration is not pathogenic. mtDNA copy number and steady-state levels of heavy- and light-strand transcripts were unaltered in clone CA 17.1. The steady-state levels of RNAs made from the two promoters (either from the heavy-strand or from the light-strand) were also similar, indicating that oppositely oriented promoters did not interfere with each other. PMID- 9199558 TI - Molecular epidemiology and diagnosis of PBG deaminase gene defects in acute intermittent porphyria. AB - Acute intermittent porphyria (AIP) is the major autosomal dominant form of acute hepatic porphyrias. The disease is due to mutations in the gene encoding for porphobilinogen (PBG) deaminase and is characterized by life-threatening neurovisceral attacks, often precipitated by drugs, fasting, cyclical hormonal changes, or infectious diseases. This report describes a prospective study on the molecular epidemiology of PBG deaminase gene defects in AIP. It uses a sensitive, reliable, and easy-to-handle method for routine AIP molecular diagnosis and family study based on an exon-by-exon denaturing gradient gel electrophoresis (DGGE) strategy followed by direct sequencing. Fifteen genomic DNA fragments, including all the coding sequence and covering 3.35 kb of the PBG deaminase gene, were investigated in 405 subjects from 121 unrelated French Caucasian AIP families who had not been screened previously at the DNA level. PBG deaminase gene mutations were identified in 109 families, but only 78 were of different type, and each of them had a prevalence rate < 5%. Among these mutations, 33 had not been published previously. Sixty percent of these 78 mutations were located in only three exons (exons 10, 12, and 14), 44% were missense, 18% were splice defect, 19% were frameshift, and 16% were nonsense. In addition, two de novo mutational events were characterized. The evaluation of the efficiency of the standard PBG deaminase enzymatic screening method for gene-carrier detection indicated 95% of concordancy with the molecular-based diagnosis. PMID- 9199559 TI - Spectrum of mutations in the OCRL1 gene in the Lowe oculocerebrorenal syndrome. AB - The oculocerebrorenal syndrome of Lowe (OCRL) is a multisystem disorder characterized by congenital cataracts, mental retardation, and renal Fanconi syndrome. The OCRL1 gene, which, when mutated, is responsible for OCRL, encodes a 105-kD Golgi protein with phosphatidylinositol (4,5)bisphosphate (PtdIn[4,5]P2) 5 phosphatase activity. We have examined the OCRL1 gene in 12 independent patients with OCRL and have found 11 different mutations. Six were nonsense mutations, and one a deletion of one or two nucleotides that leads to frameshift and premature termination. In one, a 1.2-kb genomic deletion of exon 14 was identified. In four others, missense mutations or the deletion of a single codon were found to involve amino acid residues known to be highly conserved among proteins with PtdIns(4,5)P2 5-phosphatase activity. All patients had markedly reduced PtdIns(4,5)P2 5-phosphatase activity in their fibroblasts, whereas the ocrl1 protein was detectable by immunoblotting in some patients with either missense mutations or a codon deletion but was not detectable in those with premature termination mutations. These results confirm and extend our previous observation that the OCRL phenotype results from loss of function of the ocrl1 protein and that mutations are generally heterogeneous. Missense mutations that abolish enzyme activity but not expression of the protein will be useful for studying structure-function relationships in PtdIns(4,5)P2 5-phosphatases. PMID- 9199560 TI - A rare branch-point mutation is associated with missplicing of fibrillin-2 in a large family with congenital contractural arachnodactyly. AB - Congenital contractural arachnodactyly (CCA) is an autosomal dominant disorder that is phenotypically similar to but genetically distinct from Marfan syndrome. Genetic-linkage analysis has implicated the fibrillin-2 gene (FBN2) as the CCA locus. Mutation analysis of two isolated CCA patients revealed missense mutations, indicating that defects in FBN2 may be responsible for this disorder. However, cosegregation of a mutant allele with the disease phenotype has not yet been established. We have investigated the primary cause of CCA in a large well characterized kindred with five generations comprising 18 affected individuals. Previous studies demonstrated linkage of this family's CCA phenotype to FBN2. Mutation analysis of cDNA derived from the proband and her affected brother, using a nonisotopic RNase cleavage assay, revealed the partial skipping of exon 31. Approximately 25% mutant transcript is produced, which is apparently sufficient to cause a CCA phenotype. Sequence analysis of genomic DNA revealed an unusual base composition for intron 30 and identified the mutation, a g-26t transversion, in the vicinity of the splicing branch-point site in intron 30. Genomic DNA from 30 additional family members, both affected and unaffected, then was analyzed for the mutation. The results clearly demonstrate cosegregation of the branch-point mutation with the CCA phenotype. This is the first report of a CCA mutation in a multiplex family, unequivocally establishing that mutation in FBN2 are responsible for the CCA phenotype. In addition, branch-point mutations only very rarely have been associated with human disease, suggesting that the unusual composition of this intron influences splicing stability. PMID- 9199561 TI - Identification of mutations in the duplicated region of the polycystic kidney disease 1 gene (PKD1) by a novel approach. AB - Mutation screening of the major autosomal dominant polycystic kidney disease gene (PKD1) has been complicated by the large transcript size (> 14 kb) and by reiteration of the genomic area encoding 75% of the protein on the same chromosome (the HG loci). The sequence similarity between the PKD1 and HG regions has precluded specific analysis of the duplicated region of PKD1, and consequently all previously described mutations map to the unique 3' region of PKD1. We have now developed a novel anchored reverse-transcription-PCR (RT-PCR) approach to specifically amplify duplicated regions of PKD1, employing one primer situated within the single-copy region and one within the reiterated area. This strategy has been incorporated in a mutation screen of 100 patients for more than half of the PKD1 exons (exons 22-46; 37% of the coding region), including 11 (exons 22-32) within the duplicated gene region, by use of the protein-truncation test (PTT). Sixty of these patients also were screened for missense changes, by use of the nonisotopic RNase cleavage assay (NIRCA), in exons 23-36. Eleven mutations have been identified, six within the duplicated region, and these consist of three stop mutations, three frameshifting deletions of a single nucleotide, two splicing defects, and three possible missense changes. Each mutation was detected in just one family (although one has been described elsewhere); no mutation hot spot was identified. The nature and distribution of mutations, plus the lack of a clear phenotype/genotype correlation, suggest that they may inactivate the molecule. RT-PCR/PTT proved to be a rapid and efficient method to detect PKD1 mutations (differentiating pathogenic changes from polymorphisms), and we recommend this procedure as a firstpass mutation screen in this disorder. PMID- 9199562 TI - Identification of proximal spinal muscular atrophy carriers and patients by analysis of SMNT and SMNC gene copy number. AB - The survival motor neuron (SMN) transcript is encoded by two genes, SMNT and SMNC. The autosomal recessive proximal spinal muscular atrophy that maps to 5q12 is caused by mutations in the SMNT gene. The SMNT gene can be distinguished from the SMNC gene by base-pair changes in exons 7 and 8. SMNT exon 7 is not detected in approximately 95% of SMA cases due to either deletion or sequence-conversion events. Small mutations in SMNT now have been identified in some of the remaining nondeletion patients. However, there is no reliable quantitative assay for SMNT, to distinguish SMA compound heterozygotes from non-5q SMA-like cases (phenocopies) and to accurately determine carrier status. We have developed a quantitative PCR assay for the determination of SMNT and SMNC gene-copy number. This report demonstrates how risk estimates for the diagnosis and detection of SMA carriers can be modified by the accurate determination of SMNT copy number. PMID- 9199563 TI - Haplotype and mutation analysis in Japanese patients with Wilson disease. AB - Wilson disease (WD), an autosomal recessive disorder of copper transport, is characterized by impaired biliary excretion and by impaired incorporation of copper into ceruloplasmin. Toxic accumulation of copper causes tissue damage, primarily in the liver, brain, and kidneys. The gene for WD (ATP7B) has been cloned, and the protein product is predicted to be a copper-transporting P-type ATPase with high amino acid identity with that for Menkes disease, an X-linked disorder of copper transport. Mutation screening in WD patients has led to the identification of at least 40 mutations. In addition, haplotype analysis using three dinucleotide-repeat markers, D13S314, D13S301, and D13S316, has been a useful indicator of specific mutations. We have determined haplotypes for the patients and their parents and sibs, in 21 unrelated WD families from Japan. Twenty-eight different haplotypes were observed on 42 WD chromosomes. In all the patients, the ATP7B coding sequence, including the intron-exon boundaries, was screened for mutations, by SSCP, followed by direct-sequence analysis of the shifted fragments. We identified 13 mutations, of which 11 mutations are novel, including 7 mutations-1 insertion, 4 deletions, and 2 missense mutations-in the coding region. The mutations reported in previous studies are 2299insC and Arg778Leu. Two patients were shown to have the 2299insC mutation, which has occurred in many different haplotypes in several populations, indicating a mutation hot spot. Primer-extension analysis of ATP7B mRNA has revealed multiple transcription start sites. Four of the novel mutations (three 1-bp changes and one 5-bp deletion) occur in the 5' UTR and may result in altered expression of the WD gene. PMID- 9199564 TI - Intracellular mitochondrial triplasmy in a patient with two heteroplasmic base changes. AB - We report the clinical, biochemical, and genetic investigation of a patient with a severe mitochondrial encephalomyopathy. Genetic studies identified a novel, heteroplasmic tRNA mutation at nt 10010. This T-->C transition is located in the DHU loop of mitochondrial tRNA(Gly). In skeletal muscle, it was present at lower levels in cytochrome c oxidase (COX)-normal (87.2% +/- 11%) compared with COX deficient fibers (97.3% +/- 2.6%); it was found in skin fibroblasts and blood cells, but at lower levels of heteroplasmy (15% +/- 6% and 17% +/- 10%, respectively). A second, heteroplasmic transition (A-->G), at nt 5656, showed a different distribution than the tRNA(Gly) mutation, with very low levels in skeletal muscle (< 3%) but higher levels in blood (22.7% +/- 3%) and skin fibroblasts (21% +/- 2%). These transitions were followed both in vivo, by repeat biopsy and blood sampling, and in vitro, by establishing primary cultures of myoblasts and skin fibroblasts. Repeat muscle biopsy showed a dramatic increase in COX-deficient fibers, but not of the tRNAGly mutation. Indeed, no significant change in heteroplasmy was measured for either substitution in muscle or blood. In vitro analysis gave very different results. The T10010C was not found in cultured myoblasts, even at early passage. In uncloned fibroblasts, the T10010C was stable (approximately 10%) for several passages but then gradually was lost. In contrast, the A5656G rose progressively from 27% to 91%. In cloned fibroblasts, different combinations of both base-pair changes and wild type could be identified, confirming the presence of clonal, intracellular triplasmy. PMID- 9199565 TI - Haplotype analysis of hemochromatosis: evaluation of different linkage disequilibrium approaches and evolution of disease chromosomes. AB - We applied several types of linkage-disequilibrium calculations to analyze the hereditary hemochromatosis (hh) locus. Twenty-four polymorphic markers in the major histocompatibility complex (MHC) class I region were used to haplotype hh and normal chromosomes. A total of 169 hh and 161 normal chromosomes were analyzed. Disequilibrium values were found to be high over an unusually large region beginning 150 kb centromeric of HLA-A and extending nearly 5 Mb telomeric of it. Recombination in this region was approximately 28% of the expected value. This low level of recombination contributes to the unusually broad region of linkage disequilibrium found with hh. The strongest disequilibrium was found at locus HLA-H (delta = .84) and at locus D6S2239 (delta = .85), a marker approximately 10 kb telomeric to HLA-H. All disequilibrium methods employed in this study found peak disequilibrium at HLA-H or D6S2239. The cys282tyr mutation in HLA-H, a candidate gene for hh, was found in 85% of disease chromosomes. A haplotype phylogeny for hh chromosomes was constructed and suggests that the mutation associated with the most common haplotype occurred relatively recently. The age of the hh mutation was estimated to be approximately 60-70 generations. Disequilibrium was maintained over a greater distance for hh-carrying chromosomes, consistent with a recent mutation for hh. Our data provide a reasonable explanation for previous difficulties in localizing the hh locus and provide an evolutionary history for disease chromosomes. PMID- 9199566 TI - Haplotypes of angiotensinogen in essential hypertension. AB - The M235T polymorphism of the angiotensinogen gene (AGT) has been associated with essential and pregnancy-induced hypertension. Generation of haplotypes can help to resolve whether the T235 allele itself predisposes to the development of hypertension or acts as a marker of an unknown causal molecular variant. We identified 10 diallelic polymorphisms at the AGT locus and genotyped both a series of 477 probands of hypertensive families and 364 controls, all French Caucasians, as well as a series of 92 hypertensives and 122 controls from Japan. Despite a large ethnic difference in gene frequency, a significant association of T235 with hypertension was observed both in Cancasians (.46 vs. .38, P = .004) and in Japanese (.91 vs. .76, P = .002). In both groups, the G-->A substitution located at position -6 upstream of the initial transcription site occurred at the same frequency and in complete linkage disequilibrium with the T235 allele. No other polymorphism was found to be consistently associated with hypertension. Five informative haplotypes subdividing the T235 allele were generated. Whereas two of them were associated with hypertension in Caucasians, none of these two haplotypes (H3 and H4) reached statistical significance in Japanese. The analysis of the AGT-GT repeat revealed marked linkage disequilibriums between each of the diallelic polymorphisms and some (GT)n alleles, with similar patterns in the two populations. The strong disequilibrium between M235 and (GT)16 explained the increased frequency of that particular allele in French controls compared with hypertensives (.42 vs. .36, P < .01). The haplotype combining the M235T and G-6A polymorphisms appears as the ancestral allele of the human AGT gene and as the one associated with hypertension. PMID- 9199567 TI - Homozygosity and linkage-disequilibrium mapping of the urofacial (Ochoa) syndrome gene to a 1-cM interval on chromosome 10q23-q24. AB - The urofacial (Ochoa) syndrome (UFS) is a rare autosomal recessive disease characterized by congenital obstructive uropathy and abnormal facial expression. The patients present with enuresis, urinary-tract infection, hydronephrosis, and voiding dysfunctions as a result of neurogenic bladders. To map the UFS gene, a genome screen using a combination of homozygosity-mapping and DNA-pooling strategies was performed in 20 selected patients, one patient pool, and three control pools (unaffected relatives). After analyses of 36 randomly chosen markers, D10S677 was identified as being linked to and associated with UFS, as suggested by a significant excess of homozygosity in patients compared with that in unaffected relatives (P < 10(-6)), as well as by the allelic-frequency differences between the patient pool and control pools. Ten additional markers flanking D10S677 and covering a 22-cM region then were analyzed to fine-map the UFS gene by use of haplotype (linkage disequilibrium) analysis. All 31 patients were found to be homozygous for two closely linked markers (D10S1726 and D10S198) located approximately 5 cM telomeric to D10S677, whereas only 12% of the unaffected relatives were homozygous for both markers (P < 10(-19)). Several patients are heterozygous at two markers immediately flanking D10S1726/D10S198, one on the centromeric side (D10S1433) and the other on the telomeric side (D10S603). These recombinational events place the UFS gene near D10S1726/D10S198 and within a 1-cM interval defined by D10S1433 and D10S603 on chromosome 10q23 q24. PMID- 9199568 TI - Localization of a novel X-linked progressive cone dystrophy gene to Xq27: evidence for genetic heterogeneity. AB - Clinical reexamination and DNA linkage analysis were carried out in an X-linked progressive cone dystrophy (XLPCD) family, previously described by Pinckers and Timmerman in 1981. In a large pedigree segregating XLPCD, by use of > or = 27 markers spanning the entire X chromosome, a novel locus for XLPCD was identified in Xq27. All other regions on the chromosome could be excluded. Since this novel locus is distinct from previously identified genes or regions involved in XLPCD, we further establish genetic heterogeneity underlying this disease entity. PMID- 9199569 TI - A new locus for dominant "zonular pulverulent" cataract, on chromosome 13. AB - Inherited cataract is a clinically and genetically heterogeneous disease that most often presents as a congenital autosomal dominant trait. Here we report the linkage of a new locus for dominant "zonular pulverulent" cataract (CZP) to chromosome 13. To map the CZP locus we performed molecular-genetic linkage analysis using microsatellite markers in a five-generation English pedigree. After exclusion of eight known loci and several candidate genes for autosomal dominant cataract, we obtained significantly positive LOD scores (Z) for markers D13S175 (maximum Z [Zmax] = 4.06; maximum recombination frequency [theta max] = 0) and D13S1236 (Zmax = 5.75, theta max = 0). Multipoint analysis gave Zmax = 6.62 (theta max = 0) at marker D13S175. Haplotype data indicated that CZP probably lies in the centromeric region of chromosome 13, provocatively close to the gene for lens connexin46. PMID- 9199570 TI - Lysinuric protein intolerance (LPI) gene maps to the long arm of chromosome 14. AB - Lysinuric protein intolerance (LPI) is an autosomal recessive disease characterized by defective transport of cationic amino acids and by hyperammonemia. Linkage analysis in 20 Finnish LPI families assigned the LPI gene locus to the proximal long arm of chromosome 14. Recombinations placed the locus between framework markers D14S72 and MYH7, a 10-cM interval in which the markers D14S742, D14S50, D14S283, and TCRA showed no recombinations with the phenotype. The phenotype was in highly significant linkage disequilibrium with markers D14S50, D14S283, and TCRA. The strongest allelic association obtained with marker TCRA, resulting in a P(excess) value of .98, suggests that the LPI gene locus lies in close proximity to this marker, probably within a distance of < 100 kb. PMID- 9199571 TI - Genetic mapping using microcell-mediated chromosome transfer suggests a locus for Nijmegen breakage syndrome at chromosome 8q21-24. AB - Nijmegen breakage syndrome (NBS) is an autosomal recessive disorder characterized by microcephaly, short stature, immunodeficiency, and a high incidence of cancer. Cultured cells from NBS show chromosome instability, an increased sensitivity to radiation-induced cell killing, and an abnormal cell-cycle regulation after irradiation. Hitherto, patients with NBS have been divided into the two complementation groups V1 and V2, on the basis of restoration of radioresistant DNA synthesis, suggesting that each group arises from a different gene. However, the presence of genetic heterogeneity in NBS has been considered to be controversial. To localize the NBS gene, we have performed functional complementation assays using somatic cell fusion between NBS-V1 and NBS-V2 cells, on the basis of hyper-radiosensitivity, and then have performed a genomewide search for the NBS locus, using microcell-mediated chromosome transfer followed by complementation assays based on radiosensitivity. We found that radiation resistance was not restored in the fused NBS-V1 and NBS-V2 cells and that only human chromosome 8 complements the sensitivity to ionizing radiation, in NBS cell lines. In complementation assays performed after the transfer of a reduced chromosome, merely the long arm of chromosome 8 was sufficient for restoring the defect. Our results strongly suggest that NBS is a homogeneous disorder and that the gene for NBS is located at 8q21-24. PMID- 9199572 TI - Skewed segregation of the mtDNA nt 8993 (T-->G) mutation in human oocytes. AB - Rapid changes in mtDNA variants between generations have led to the bottleneck theory, which proposes a dramatic reduction in mtDNA numbers during early oogenesis. We studied oocytes from a woman with heteroplasmic expression of the mtDNA nt 8993 (T-->G) mutation. Of seven oocytes analyzed, one showed no evidence of the mutation, and the remaining six had a mutant load > 95%. This skewed expression of the mutation in oocytes is not compatible with the conventional bottleneck theory. A possible explanation is that, during amplification of mtDNA in the developing oocyte, mtDNA from one mitochondrion is preferentially amplified. Thus, subsequent mature oocytes may contain predominantly wild-type or mutant mitochondrial genomes. PMID- 9199573 TI - Heritability of longitudinal changes in coronary-heart-disease risk factors in women twins. AB - Numerous studies have demonstrated genetic influences on levels of coronary heart disease (CHD) risk factors, but there also may be genetic effects on the intraindividual variation in these risk factors over time. Changes in risk factors are likely to reflect genetic-environmental interactions and may have important implications for understanding CHD risk. The present study examines the heritability of changes in CHD risk factors, using data from the two examinations by the Kaiser Permanente Women Twins Study, performed a decade apart. The sample consisted of 348 pairs of women twins who participated in both examinations, including 203 MZ pairs and 145 DZ pairs. Average ages at the two examinations were 41 and 51 years, respectively. By means of three different statistical analytic approaches, moderate heritability estimates were demonstrated for changes in LDL cholesterol (h2 = .25-.36) and in HDL cholesterol (h2 = .23-.58), some of which were statistically significant. Although small to moderate heritability estimates were found for systolic blood pressure (.18-.37; P < .05 for some estimates), no genetic influence on changes in diastolic blood pressure was detected. Based on longitudinal twin data in women, this study demonstrates a genetic influence on changes in both lipoprotein risk factors and systolic blood pressure over a decade. In addition to environmental factors, which clearly are operating, the effect of various "variability genes" may be acting independently of the genetic influences on the absolute levels of these risk factors. Both mapping the gene(s) underlying intraindividual variations in these CHD risk factors and understanding their function(s) could lead to targeted intervention strategies to reduce CHD risk among genetically susceptible individuals. PMID- 9199574 TI - Fine-scale genetic mapping based on linkage disequilibrium: theory and applications. AB - Linkage-disequilibrium mapping (LDM) recently has been hailed as a powerful statistical method for fine-scale mapping of disease genes. After reviewing its historical background and methodological development, we present a general, mathematical, and conceptually coherent framework for LDM that incorporates multilocus and multiallelic markers and mutational processes at the marker and disease loci. With this framework, we address several issues relevant to fine scale mapping and propose some efficient computational methods for LDM. We implement various LDM methods that incorporate population growth, recurrent mutation, and marker mutations, on the basis of a general framework. We demonstrate these methods by applying them to published data on cystic fibrosis, Huntington disease, Friedreich ataxia, and progressive myoclonus epilepsy. Since the genes responsible for these diseases all have been cloned, we can evaluate the performance of our methods and can compare ours with that of other methods. Using the proposed methods, we successfully and accurately predicted the locations of genes responsible for these diseases, on the basis of published data only. PMID- 9199575 TI - The Val985Met insulin-receptor variant in the Danish Caucasian population: lack of associations with non-insulin-dependent diabetes mellitus or insulin resistance. PMID- 9199576 TI - Absence of mutations raises doubts about the role of the 70-kD peroxisomal membrane protein in Zellweger syndrome. PMID- 9199577 TI - Disease relevance of the so-called secondary Leber hereditary optic neuropathy mutations. PMID- 9199578 TI - A mutation in the MTM1 gene invalidates a previous suggestion of nonallelic heterogeneity in X-linked myotubular myopathy. PMID- 9199579 TI - Detection of an atypical 22q11 deletion that has no overlap with the DiGeorge syndrome critical region. PMID- 9199580 TI - Linkage disequilibrium between the spinocerebellar ataxia 3/Machado-Joseph disease mutation and two intragenic polymorphisms, one of which, X359Y, affects the stop codon. PMID- 9199581 TI - A common mtDNA polymorphism associated with variation in plasma triglyceride concentration. PMID- 9199582 TI - Up-regulation of the brain and Purkinje-cell forms of dystrophin transcripts, in Becker muscular dystrophy. PMID- 9199583 TI - Bilateral retinoblastoma in a male patient with an X; 13 translocation: evidence for silencing of the RB1 gene by the spreading of X inactivation. PMID- 9199584 TI - Meiotic drive at the myotonic dystrophy and the cone-rod dystrophy loci on chromosome 19q13.3. PMID- 9199585 TI - Reply to letter by Dr. Remmelink. PMID- 9199586 TI - Notes on recent Notes from the Clinic. PMID- 9199587 TI - Single-phase versus two-phase treatment: differing but ethical opinions. PMID- 9199588 TI - Comparison of the effectiveness of two types of toothbrushes on the oral hygiene of patients undergoing orthodontic treatment with fixed appliances. AB - The purpose of this study was to investigate whether orthodontic toothbrushes were superior to classical toothbrushes in the elimination of microbial dental plaque on teeth and brackets and in the maintenance of periodontal tissue health in patients, ages 12 to 22 years, with fixed appliances. Twenty patients undergoing orthodontic treatment with fixed appliances and brushing with the Bass technique were included in the study. Ten patients used the Oral B Ortho type toothbrushes (Oral B Laboratories Ltd.), whereas the remaining 10 patients used the Oral B Plus 35 type toothbrushes. Quigley-Hein plaque index, bonded bracket index, sulcus bleeding index, and periodontal pocket depth measurements were made at the beginning of the study and a month later. No statistically significant difference was found for plaque, sulcus bleeding, and periodontal pocket depth between Oral B Ortho and Plus 35 groups when the preinvestigatory and postinvestigatory measurements for the vestibular and proximal surfaces of upper and lower teeth were compared. This short-term study concluded that the Ortho type toothbrush is not superior to the Plus 35-type toothbrush. PMID- 9199589 TI - Salivary nickel and chromium in subjects with different types of fixed orthodontic appliances. AB - The aim was to investigate nickel and chromium concentrations in saliva of patients with different types of fixed appliances. Saliva samples were collected from 47 orthodontic patients, ages 8 to 30 years. Four samples from each subject were collected: (1) before insertion of the appliance, (2) 1 to 2 days after, (3) 1 week after, and (4) 1 month after insertion of the appliance. A considerable variation in the concentrations of both nickel and chromium was observed. No significant differences were found between the no-appliance samples and the samples obtained after insertion of the appliances. The results suggest that nickel and chromium concentrations of saliva are not significantly affected by fixed orthodontic appliances during the first month of treatment. PMID- 9199590 TI - Effect of pumice prophylaxis on the bond strength of orthodontic brackets. AB - Pumice prophylaxis has long been accepted as a prerequisite for achieving adequate enamel etching during orthodontic bonding procedures. Three methods were used in this study to examine the effects of pumice prophylaxis on the bond strength of orthodontic brackets: (1) shear bond strength of brackets that were bonded to extracted premolars after surface preparation procedures, which either included or did not include prior pumice prophylaxis, was evaluated; (2) scanning electron microscopy (SEM) was used to examine the surface characteristics of teeth that had been etched with and without prior pumice prophylaxis; and (3) rate of bracket failure in patients who had had brackets bonded with and without prior pumice prophylaxis was recorded during an average treatment time of 18 months. No significant differences were noted in bond strength, general etched enamel surface characteristics, or bracket retention rates. Some specific differences, however, were noted on SEM in localized areas of the etched enamel surfaces, although these did not appear to affect the bond strength or bracket retention rates ultimately attained. PMID- 9199591 TI - Clinical effects of chlorhexidine mouthwashes on patients undergoing orthodontic treatment. AB - This study compared the short-term clinical effect of 0.12% chlorhexidine gluconate and placebo mouthrinses in 30 adolescents (ages 11 to 15) undergoing orthodontic treatment. Subjects were randomized into experimental (CHX) and control (C) groups. Baseline values were recorded 10 days after prophylaxis and included Plaque index (PI), Gingival index (GI), Rentention Index (RI), Discoloration index (DI), and probing depths (PD). Both groups (CHX and C) received soft toothbrushes with instructions to brush twice daily, as well as the CHX and placebo mouthrinses, respectively, with oral and written instructions for rinsing twice daily with 15 ml for 30 seconds. Reevaluations were performed 1, 2, and 3 months after baseline, except for the DI and PD, which were only assessed at 3 months. The Student's t test and the paired t test were used to analyze the data at the P < 0.05 level of significance. No differences between groups were seen at baseline for any of the parameters. At 30 days, there was a significant difference for the RI between CHX (0.15 +/- 0.16; mean +/- SD) and C (0.05 +/- 0.06) at the mesial buccal, and for CHX (0.07 +/- 0.10) and C (0.02 +/- 0.05) at the midbuccal. The 60-day evaluation showed similar results. At 90 days, lower PI were observed in the CHX group at the distal buccal (0.38 +/- 0.19), midbuccal (0.22 +/- 0.17), and mesial buccal (0.47 +/- 0.22) sites as compared with the C group (0.97 +/- 0.38, 0.83 +/- 0.40, and 0.95 +/- 0.43, respectively). A similar trend was noted with the GI, as the lower values were related to the CHX group. The changes of the PI and GI, at 30, 60, and 90 days, as analyzed by the paired t test, were statistically significant in the case of the experimental group, as the changes in the means were a reflection of significantly lower scores observed in the experimental group. After 3 months, the DI showed higher scores in the experimental group as compared with the control, but they were not statistically significant. Deeper PD were detected in the C group at 90 days, and they were statistically significant, except for the midlingual site. The RI did not show significant differences at 90 days, but higher values were recorded in the CHX group. The data indicate that the use of the CHX, in addition to regular oral hygiene habits, was effective in reducing plaque and gingivitis in adolescents undergoing orthodontic treatment. PMID- 9199592 TI - Forces produced by lip bumpers on mandibular molars. AB - The purpose of this study was to measure the forces produced by a lip bumper on the mandibular permanent first molars. The forces in a sample of 38 patients were measured bilaterally with specially designed gauges at rest but with their lips lightly touching, speaking the words church, phone, and pop, and swallowing water. Forces were compared between two types of lip bumpers, i.e., wire or shield, and between various anteroposterior and vertical positions of the lip bumper. The resting forces produced by the wire lip bumper 2 mm anterior to the incisors and vertically positioned at the middle of the incisor crown were 5.93 +/- 4.84 gm for the left side and 4.66 +/- 4.8 gm for the right. The forces were found to be significantly higher when the wire lip bumper was placed 4 mm anterior to the incisors and at a more gingival position, measuring 16.68 +/- 8.7 gm for the left side and 13.88 +/- 8.28 gm for the right. The shield lip bumper had higher forces both at the center of the incisor as well as when it was positioned gingivally. A large individual variation was observed. There were no statistically significant differences in force levels between male and female subjects. Speaking the words church, pop, and phone, produced forces between 11 and 23 gm, using a wire lip bumper. Swallowing produced the highest forces, between 32 and 36 gm. Lip thickness and height did not appear to affect the force levels. PMID- 9199593 TI - A study of holographic interferometry on the initial reaction of maxillofacial complex during protraction. AB - Most extraoral appliances used for protracting small or retropositioned maxilla do not allow for variations in the point of force application or in its direction. This variation may be necessary to control vertical, anteroposterior, as well as transverse effects. The purpose of this study was to investigate the initial reaction of the maxillofacial complex according to force magnitude, force direction, and point of force application. For this purpose, an antenna-type modified protraction headgear was tested with double exposure holographic interferometry on a dry human skull with well-aligned upper teeth. Fringe patterns of each protraction condition were compared and analyzed. In most cases, upward rotation of the anterior portion of the maxilla changed to translation, or to downward rotation, as force direction was changed from parallel to the occlusal plane to 20 degrees downward to the occlusal plane. Furthermore, a 500 gm force applied 15 mm above and directed 20 degrees below the occlusal plane produced a translation of the maxillary complex, indicated by a typical circular fringe pattern on the holographic plate, which represents the center of resistance of the maxilla. In most cases, with all force variables tested, a protraction of the maxilla with palatal expansion was more effective in producing translation of the maxilla than was protraction without palatal expansion. By varying force magnitude, force direction and point of force application with maxillary protraction, the amount of maxillary rotation and translation might be controlled. PMID- 9199594 TI - Incomplete canine transposition and maxillary central incisor impaction--a case report. PMID- 9199595 TI - Combined orthodontic-orthognathic surgical treatment of a Class II, Division I malocclusion. AB - This case report shows the need to extract four first premolars in addition to orthognathic surgery, even though the initial treatment plan involved a nonextraction strategy. The extractions were necessary to reduce maxillary dental protrusion and proclination and also to recover from the mandibular incisor proclination that occurred as a consequence of leveling the mandibular arch. PMID- 9199596 TI - The demographics of Dr. Geoffrey Walker's cephalometric collection. AB - The Bolton-Brush Growth Study Center (BBGSC) at Case Western Reserve University recently acquired the radiographic collection of Geoffrey F. Walker, an orthodontist, anthropologist, and pioneer computer expert. Dr. Walker's culturally diverse collection on more than 1500 persons has added lateral and frontal cephalograms to the Bolton-Brush collection. In contrast to the longitudinal Bolton-Brush study, the Walker collection contains predominantly cross-sectional samples of various tribes and ethnic groups around the world. A computerized index (similar to the Bolton-Brush index) has been created for efficient access. The addition of this unique collection enhances the BBGSC's ability to serve as a resource for orthodontic students, researchers, and practitioners, as well as the physical anthropology community. PMID- 9199597 TI - Soldered implant attachments. PMID- 9199598 TI - A new approach to indirect bonding using light-cure composites. AB - Indirect bonding offers many advantages over direct bonding. The ability to better visualize teeth for proper bracket placement and reduced chair time leads to a less stressful bonding experience for the orthodontist. Conventional approaches to indirect bonding have relied on transfer trays made of materials that are not transparent. After these trays are placed on teeth, one cannot ensure proper seating of the tray and, because of their opaque nature, the orthodontist must use self-cure composites that are very difficult to remove around the brackets after the tray is removed. A new approach has been proposed in this article, which significantly modifies the fabrication of the indirect bonding transfer tray and provides direct visualization and access to the brackets during both the laboratory and clinical stages of the procedure. Furthermore, the orthodontist may use light-cure composite resins, clean off excess composite around the brackets, and apply light curing when fully satisfied with bracket position and hygiene. PMID- 9199599 TI - Judging a digital imaging system. PMID- 9199600 TI - Litigation, legislation, and ethics. Revisiting restrictive covenants. PMID- 9199601 TI - Prognostic significance of Bcl-2 expression in localized squamous cell carcinoma of the head and neck. AB - This study was conducted to determine whether Bcl-2 overexpression in localized squamous cell carcinoma of the head and neck (SCCHN) might serve as a marker for tumors unlikely to respond to standard treatment. Tissue samples from 33 patients undergoing surgery or irradiation for early-stage SCCHN during the year 1977 to 1992 were stained for Bcl-2. All patients had either T1N0 lesions of the oral cavity, pharynx, or larynx or T1N0 or T2N0 lesions of the true vocal cords. Of the 33 patients, 26 remained disease-free after at least 3 years of follow-up; the remaining 7 patients developed either tumor recurrence or a second primary tumor, 4 of which were fatal. Twelve patients had tissue specimens staining positive for Bcl-2; 6 of these patients had a poor outcome, and 6 had a good outcome. The relationship between poor outcome and overexpression of Bcl-2 in tumor cells was statistically significant (p = .0047 by Fisher's exact test). For tumors overexpressing Bcl-2, there was no significant difference in recurrence rate between those undergoing surgery and those undergoing radiotherapy as the primary mode of treatment. The overexpression of Bcl-2 in early lesions in this study predicted a cure rate of 50%, as opposed to the generally expected 90%, suggesting that Bcl-2 is a significant prognostic indicator in early SCCHN. Future studies will determine if altering the treatment will improve outcome in these patients. PMID- 9199602 TI - Method and clinical results of a new transthyrotomy closure of the supraglottic larynx for the treatment of intractable aspiration. AB - A new procedure has been developed to surgically separate the pharynx from the trachea that employs the best features of the Montgomery technique, but restricts the closure to only the epiglottis and the aryepiglottic folds. The petiole of the epiglottis is plicated to the false vocal folds and the interarytenoid mucosa. It is performed entirely through a midline thyrotomy approach and avoids injury to any of the structures within the rima glottidis. It has been successfully performed on seven very ill patients to date. The surgical decision making process involved, a complete description of the surgical procedure, and a summary of the patients' preoperative condition, workup, and outcomes are presented and discussed. PMID- 9199603 TI - Diagnosis and management of left main stem bronchus compression. AB - There are four major variants of congenital vascular tracheal compression: innominate artery, aberrant subclavian, aorta or aortic arch anomaly, and pulmonary artery sling. These forms of vascular compression typically involve the trachea and/or the right main stem bronchus. We present eight cases of congenital vascular compression involving the left main stem bronchus. These cases represent a poorly understood variant of vascular tracheal compression. This variant represents approximately 10% of our pediatric tracheobronchial compression or stenosis patients. The finding, both noted endoscopically and now illustrated by magnetic resonance imaging, is caused by compression of the left main stem bronchus between the descending aorta and a portion of the pulmonary artery. Frequently, the descending aorta is in an abnormal anterior position with relation to the thoracic spine. Recognition of this entity is important in our experience and has influenced clinical management. In four of eight children, it required a surgical procedure directed toward the relief of the left main stem compression. PMID- 9199604 TI - Marrow-mesenchyme connections in the fetal and newborn tympanum. A new entity. AB - Examinations of 41 human fetal, 8 infant, and 8 juvenile temporal bones prepared for light microscopic evaluation revealed direct connections between the hematopoietic bone marrow and the unresolved mesenchyme in the middle ear. The connections first appeared at 15 weeks of gestation and became bridged by fibrous tissue, in most cases, by the postpartum age of 10 months. Between 16 and 18 months after birth, the marrow-mesenchyme connections gradually disappeared. The areas in which the connections were most numerous were the anterior epitympanum, the sinus tympani medial to the stapedius muscle, and transitory bone that occupies the area that will become the aditus of the antrum. Immunohistochemical staining demonstrated the existence of mature leukocytes in these connections. These connections may help protect the middle ear against bacterial invasion during the postnatal period. PMID- 9199605 TI - Giant cholesteatoma of the external auditory canal. AB - Cholesteatomas are found almost exclusively in the middle ear and mastoid. Occasionally this disease is seen in the external auditory canal. Cholesteatoma of the external auditory canal is a rare condition. Severe pain and profuse discharge associated with a normal eardrum and normal hearing are essential clinical features. In addition, we found facial paresis and conductive hearing loss in our case. Smaller cholesteatomas can be managed by frequent debridement in the office; larger lesions require surgical intervention. Surgery is successful in resolving otorrhea and relieving pain. In addition, our own experience has shown that surgery is successful in relieving facial paresis. PMID- 9199606 TI - Decrease in immunoreactive neutral endopeptidase in uvula epithelium of patients with obstructive sleep apnea. AB - The purpose of this study was to determine whether neutral endopeptidase (NEP; EC3.4.24.11) is decreased in the uvula epithelium of patients with obstructive sleep apnea (OSA). Tissues were obtained by uvulopharyngopalatoplasty in seven patients with moderate OSA and by autopsy in five individuals not known to have OSA. Using antisera to human NEP and immunoperoxidase staining, we found that NEP was localized in uvula epithelial cells of both patients with OSA and controls. However, there was a significant decrease in the number of epithelial cells staining for NEP in patients with OSA relative to controls (67 +/- 10 cells versus 261 +/- 33 cells, in 5 randomly selected high-power microscopic fields, respectively; mean +/- SEM; p < .05). The intensity of staining for NEP was similar in both groups. We conclude that immunoreactive NEP is significantly decreased in the uvula epithelium of patients with OSA. PMID- 9199607 TI - Middle ear pressure and dysfunction of the labyrinth: is there a relationship? AB - Relationships between middle ear pressure and non-infection-related cochleovestibular dysfunction have been suggested by several authors. According to some data, vertiginous attacks can be prevented by the insertion of a ventilation tube in patients suffering from Meniere's syndrome. The aim of our study was to investigate if the incidence of eustachian tube malfunction and pathologic middle ear pressure is frequent, and if routine implantation of ventilation tubes is reasonable in ears with dysfunctions of the labyrinth, including clinical Meniere's syndrome. So, we determined in our pressure chamber all active and passive parameters of eustachian tube function in 40 patients suffering from Meniere's syndrome, sudden sensory hearing impairment (SSHI), or vestibular neuronitis. Our results disclosed no nonrandom incidence of impaired tubal function among our patients compared to healthy control subjects. Pressure equalization was sufficient in most patients suffering from clinical Meniere's syndrome, and only one patient with vestibular neuronitis presented with a patulous tube. Our results show that impairment of vestibular or cochlear function is not regularly accompanied by eustachian tube dysfunction. Furthermore, no patient reported symptoms while pressure variation was performed. We conclude that variation of middle ear pressure does not usually play a role in the genesis of Meniere's syndrome, vestibular neuronitis, or SSHI. Thus, from our data, we cannot recommend routine implantation of tympanic ventilation tubes in patients suffering from Meniere's syndrome, vestibular neuronitis, or sudden hearing loss. PMID- 9199608 TI - Significant correlation between symptom score and IgG4 antibody titer following long-term immunotherapy for perennial allergic rhinitis. AB - Although there is evidence of some measure of clinical benefit as well as immunologic change during the early phase of immunotherapy, a sustained clinical response is only possible with prolonged therapy. Immunotherapy has to be administered for about 3 to 5 years for such sustained clinical efficacy. This study aimed at investigating the dynamics of IgE and IgG4 antibodies after more than 5 years of immunotherapy, to examine the statistical correlation between these antibodies and symptom scores. Our study demonstrated that the allergen specific IgE antibody level significantly decreases and the IgG4 antibody level significantly increases following immunotherapy. However, the percent decrease in IgE antibodies did not correlate with the percent decrease in symptom scores. On the other hand, the percent increase in IgG4 antibodies correlated with the percent decrease in symptom scores. We infer that an elevation of IgG4 antibodies is not simply an epiphenomenon unrelated to the underlying working mechanism of clinically successful immunotherapy, but probably makes an active contribution to symptom relief. PMID- 9199609 TI - Fine three-dimensional structure of pericytes in the vocal fold mucosa. AB - An investigation was carried out to determine the fine three-dimensional structure of pericytes in excised human vocal fold mucosa, by means of scanning and transmission electron microscopic observation. The results are summarized as follows. 1) There were many pericytes around the true capillaries, arterial capillaries, and venous capillaries in the vocal fold mucosa. 2) Newborns had pericytes around the capillaries in the vocal fold mucosa. 3) The pericytes had bulged fusiform or polygonal cell bodies and branching processes. The branching processes consisted of long and relatively thick longitudinal ones and short circumferential ones. 4) The cell body and processes of the pericytes encircled the capillaries, and the tips of the processes formed intercellular tight junctions with endothelial cells and made a firm connection with them. 5) The pericytes had many cytoplasmic filaments. 6) The pericytes in the vocal fold mucosa appeared to support and protect capillary walls in the vibrating tissue. PMID- 9199610 TI - Pressure gradients affecting the labyrinth during hypobaric pressure. Experimental study. AB - Hypobaric effects on the perilymph pressure were investigated in 18 cats. The perilymph, tympanic cavity, cerebrospinal fluid, and systemic and ambient pressure changes were continuously recorded relative to the atmospheric pressure. The pressure equilibration of the eustachian tube and the cochlear aqueduct was studied, as well as the effects of blocking these channels. During ascent, the physiologic opening of the eustachian tube reduced the pressure gradients across the tympanic membrane. The patent cochlear aqueduct equilibrated perilymph pressure to cerebrospinal fluid compartment levels with a considerable pressure gradient across the oval and round windows. With the aqueduct blocked, the pressure decrease within the labyrinth and tympanic cavities was limited, resulting in large pressure gradients toward the chamber and the cerebrospinal fluid compartments, respectively. We conclude that closed cavities with limited pressure release capacities are the cause of the pressure gradients. The strain exerted by these pressure gradients is potentially harmful to the ear. PMID- 9199611 TI - Extracellular fluid pathway inside the facial nerve fascicles. AB - The extracellular fluid pathway in the facial nerve and the diffusion of a tracer from the facial nerve to other cranial nerves was examined in the rabbit. Sodium fluorescein solution was injected into either the facial nerve fascicles or the epineurial connective tissue as a tracer at the stylomastoid foramen and then localized by fluorescence microscopy. In the facial nerve, fluorescence was observed in the endoneurium and external nerve sheath (epineurium and perineurium) through the geniculate ganglion following injection into the nerve fascicles. The vestibular, trigeminal, and glossopharyngeal ganglia also showed fluorescence on the injection side in ganglion cells and intercellular connective tissues. The results suggested that the endoneurial connective tissue constitutes a diffusion pathway inside the facial nerve fascicles and that the extracellular fluid pathway from the facial nerve to these cranial ganglia may be related to the neural spread of inflammation or neoplastic metastasis. PMID- 9199612 TI - End-to-side neurorrhaphy resulting in limited sensory axonal regeneration in a rat model. AB - This study evaluated reinnervation of an end-to-side neurorrhaphy and the resultant functional recovery in a rat model. The cut distal posterior tibial nerve was repaired to the side of an intact peroneal nerve. In one group, the epineurium of the peroneal nerve was left intact; in another group, the epineurium was stripped; in the third experimental group, a perineurial slit was created. Evaluations included walking track analysis, nerve conduction studies, muscle mass measurements, retrograde nerve tracing, and histologic evaluation. Walking tracks indicated poor functional recovery. No significant difference in nerve conduction between the experimental and control groups was seen. Gastrocnemius muscle mass measurements revealed no functional recovery in the end to-side groups. Retrograde nerve tracing revealed minimal staining of motor neurons. However, sensory neuronal staining of the dorsal root ganglia occurred in all groups. Histology revealed minimal myelinated axonal regeneration. These results suggest that predominantly sensory neural regeneration occurs in an end to-side neurorrhaphy at an end point of 16 weeks. PMID- 9199613 TI - Application of oxygen free radical scavengers to diminish the occurrence of myringosclerosis. AB - The present study was designed to establish whether or not an increased production of oxygen-derived free radicals is involved in the causation of myringosclerosis. Sclerotic lesions in the tympanic membrane were experimentally elicited by keeping rats with perforated tympanic membranes in an atmosphere containing roughly 40% oxygen. The animals were treated daily with a solution containing either copper zinc-superoxide dismutase plus catalase, deferoxamine, or copper sulfate plus iron chloride, applied to the traumatized area. After 1 week the extension of myringosclerotic plaques was determined otomicroscopically. The pars tensa and pars flaccida were then dissected free and prepared for light microscopic studies. The results showed that treatment with copper zinc superoxide dismutase plus catalase and deferoxamine inhibited or reduced the development of myringosclerosis, whereas the ears treated with copper sulfate plus iron chloride appeared unaffected. Consequently, the findings support the hypothesis that the formation of oxygen free radicals contributes significantly to the development of myringosclerosis. PMID- 9199614 TI - Melanoma of the petrous apex of the temporal bone. PMID- 9199615 TI - Monolateral aplasia of the parotid gland. PMID- 9199616 TI - Head and neck Langerhans cell histiocytosis. AB - Among the potential sites of involvement by Langerhans cell histiocytosis (LCH), the head and neck region is the most commonly cited. Though principally a pediatric disease, LCH can affect any age group. It can be unifocal (skeletal) or multifocal (skeletal and/or visceral); it appears as though the presence of visceral lesions is more common in the youngest patients, and may be associated in some with a rapidly progressive course resulting in death. Head and neck manifestations may mimic such varied entities as eczema, otitis media, osteomyelitis, and cholesteatoma. Current approaches to therapy are less aggressive than they were in the past, and are particularly intended to monitor for and treat any complicating secondary infections (which may develop in the youngest patients with multifocal disease including visceral involvement). The prognosis is very good for unifocal skeletal system disease, and poor for multifocal disease with involvement of tissues other than bone. PMID- 9199617 TI - Pathologist's shoulder. PMID- 9199618 TI - Pathologist's shoulder. PMID- 9199619 TI - Interinstitutional comparison of frozen section turnaround time. A College of American Pathologists Q-Probes study of 32868 frozen sections in 700 hospitals. AB - OBJECTIVES: To study the intraoperative turnaround time for performing a frozen section (FS) and to examine pathology practice variables that influence it. DESIGN: Over a 4-month period in 1995, participants in the College of American Pathologists Q-Probes laboratory quality improvement program prospectively collected data on up to 30 FS procedures performed on elective inpatient surgical cases and completed questionnaires profiling their FS practice characteristics. SETTING: Surgical pathology laboratories serving private and public hospitals. PARTICIPANTS: Seven hundred institutions located in North America (667), Australia (12), New Zealand (1), the United Kingdom (3), Hong Kong (1), Mexico (1), and Norway (1). MAIN OUTCOME MEASURES: The 90% FS block completion time defined as the time interval, in minutes, within which the fastest 90% of all FS blocks were completed, measured from the time pathologists received FS specimens to the time they communicated FS results to the surgeon. RESULTS: Participants submitted data on 32868 FS blocks. Ninety percent of FS procedures were completed within 20 minutes. Frozen section turnaround times exceeding 20 minutes, termed outlier turnaround times, were more likely to occur when more than one pathologist participated in the FS diagnosis, pathology residents and medical students participated in the FS procedure, the pathologist had to retrieve and review previous case material during the FS procedure, the pathologist simultaneously received additional specimens from other FS cases, the pathologist was unable to reach a final FS diagnosis, and when technical problems occurred during the FS procedure. Seventy percent of all participating hospitals completed 90% of their frozen sections within 20 minutes. The institutional 90% completion times were shorter for hospitals containing 300 or fewer occupied beds than for those containing more than 300 occupied beds. CONCLUSIONS: The data suggest that 90% of FS block turnaround times can be performed within 20 minutes, measured from the time that pathologists receive FS specimens to the time that pathologists return FS diagnoses to surgeons. PMID- 9199620 TI - The serum anion gap. Has the reference interval really fallen? AB - OBJECTIVE: To compare the anion gap (calculated as the sodium concentration minus the sum of the chloride and total carbon dioxide concentrations) reference interval for three automated chemistry analyzers. DESIGN: We measured serum sodium, chloride, and total carbon dioxide on aliquoted specimens using three commercial instruments. Quality control and proficiency survey materials were run to ensure that the analyzers were functioning optimally. SETTING: Three separate clinical laboratories affiliated with one university medical center participated in the study. PARTICIPANTS: Healthy volunteers from 20 to 60 years of age were recruited from within a clinical laboratory. MAIN OUTCOME MEASURES: The mean and standard deviations of the anion gaps measured by each method were calculated. RESULTS: The parametric reference intervals (+/-2 SD from the mean) were 5 to 10 mmol/L for the Beckman Synchron CX3 analyzer, 9 to 14 mmol/L for the Boehringer Mannheim Hitachi 717 analyzer, and 8 to 13 mmol/L for the Johnson & Johnson Vitros 950 analyzer. CONCLUSIONS: Our results suggest that while it may be appropriate to lower the anion gap reference interval to 5 to 10 mmol/L for some analyzers, as suggested by earlier reports, 9 to 14 mmol/L may be a more appropriate reference interval for other analyzers. For the anion gap to be an effective tool for diagnosing acid-base disorders, clinical laboratorians need to establish (or at least verify) the anion gap reference interval for the instrumentation used in their laboratory, inform clinicians of this reference interval, and perform quality control studies to ensure that the reference interval for this calculated result remains valid. PMID- 9199621 TI - Automated five-part white blood cell differential counts. Efficiency of software generated white blood cell suspect flags of the hematology analyzers Sysmex SE 9000, Sysmex NE-8000, and Coulter STKS. AB - OBJECTIVE: The present study evaluates the efficiency of software-generated white blood cell (WBC) "suspect flags" of the hematology analyzers Sysmex SE-9000, Sysmex NE-8000, and Coulter STKS. DESIGN: Automated WBC differential counts were considered positive if they contained any suspect WBC flag indicating the presence of blasts, myeloid precursor cells, or abnormal lymphocytes. Reference differential counts were performed by microscopic examination of 400 WBCs per sample. After comparison to the reference method, automated differential counts were classified as true-positive, true-negative, false-positive, and false negative. The flagging efficiency of analyzers was expressed as a percentage of subjects correctly classified. SPECIMENS: Four hundred sixty-seven blood samples were randomly chosen for comparison analysis from various inpatient and outpatient departments of the Vienna university hospital, Austria. RESULTS: The efficiency rates of flagging for the presence of > or = 1% abnormal WBCs were 78% (SE-9000), 77% (NE-8000), and 72% (Coulter STKS). The flagging efficiencies were best for samples with normal WBC counts. With regard to the specific suspect flags, the flagging of blast cells was most efficient on all analyzers. CONCLUSIONS: Our results demonstrate the comparable overall performance of three analyzers, SE-9000, NE-8000, and Coulter STKS. They further underscore the importance of critical interpretation of automated differential counts, because at a detection limit of > or = 1% abnormal WBCs > 20% of samples were not correctly flagged by either analyzer. PMID- 9199622 TI - Osteopontin and p53 expression are associated with tumor progression in a case of synchronous, bilateral, invasive mammary carcinomas. AB - OBJECTIVE: To examine the association between expression of osteopontin (OPN), p53, other molecular markers (Ki-67, c-erb B2, and estrogen receptor protein) and tumor progression in a case of synchronous, bilateral, invasive mammary carcinomas of the same histology. DESIGN: Immunohistochemistry was performed on formalin-fixed, paraffin-embedded tissue sections. Plasma OPN level was determined by a quantitative antigen capture assay. SETTING: The patient was seen, treated, and followed up for a period of 5 years at the London Regional Cancer Centre, Ontario, Canada. PATIENT: A 60-year-old woman presented with bilateral infiltrating mammary carcinomas of the same histologic type and grade. Bilateral mastectomy and axillary node dissection showed involvement of 3 of 12 right axillary and 0 of 11 left axillary lymph nodes. She later developed a right chest wall recurrence, followed by widespread metastatic disease to the skull, liver, and left femur. RESULTS: The primary tumor of the right breast was OPN- and p53-positive, whereas the tumor of the left breast was negative for both markers. The development of right axillary lymph node metastases, chest wall recurrence, and distant metastases was associated in all instances with an immunohistochemical profile of high level expression of OPN and p53. Plasma assay for OPN at the time of last admission showed a markedly elevated OPN level. CONCLUSIONS: Increased p53 expression was found to be associated with increased tumor aggressiveness. The association of increased OPN expression with increased malignancy in breast cancer is a novel finding and raises the possibility of a role for OPN in tumor progression, as well as the potential for this marker in predicting clinical aggressiveness. PMID- 9199623 TI - Multiparameter analysis of DNA content and cytokeratin expression in breast carcinoma by laser scanning cytometry. AB - OBJECTIVE: The objective of this study was to test a new laboratory technology, laser scanning cytometry, for the purpose of performing multiparameter DNA content analysis of breast carcinomas. DESIGN: We developed a simplified method of multiparameter DNA content analysis using cytokeratin expression to positively gate epithelial cells. Over 300 consecutive cases of breast carcinoma were analyzed by multiparameter laser scanning cytometry. The first 73 cases were analyzed in parallel by single parameter flow cytometry. SETTING: The Department of Pathology, Christ Hospital and Medical Center, Oak Lawn, Ill. SPECIMENS: Three hundred eighteen consecutive cases of breast carcinoma presenting between March 1994 and December 1995. MAIN OUTCOME MEASURES: Outcome measures included the percentage of cases for which DNA content analysis could be successfully performed given the limitations of specimen size. Additionally, for the first 73 cases, laser scanning cytometry results were compared with flow cytometry results. RESULTS: All of the first 73 cases were successfully analyzed by laser scanning cytometry, but for 8 cases (11%) there was insufficient material for flow cytometry. Correlation of DNA content for the remaining 65 cases analyzed in parallel by the two methods was nearly perfect (p = .994). Five seemingly discrepant cases highlighted the importance of cytokeratin gating of epithelial cells by any technique, as well as other advantages specific to laser scanning cytometry, such as the ability to examine individual cells microscopically and correlate cytologic morphology with DNA content results. CONCLUSIONS: Laser scanning cytometry is a promising new technology for DNA content analysis of solid tissue tumors. Further work needs to be performed to validate the prognostic potential of the laser scanning cytometric assay results and to generate methodologies aimed at providing highly objective determinations of tumor cell S-phase fraction. PMID- 9199624 TI - Intraluminal crystalloids in breast carcinoma. Immunohistochemical, ultrastructural, and energy-dispersive x-ray element analysis in four cases. AB - OBJECTIVE: Intraluminal crystalloids have been described in the prostate, salivary gland, and ovary, but have not yet been reported in the breast. We report four cases of breast carcinoma in which these crystalloids were found in ducts with intraductal carcinoma or atypical hyperplasia. The presence of intraluminal crystalloids may be a useful adjunct in making a diagnosis of carcinoma or may be a feature to look for as a marker for the presence of carcinoma. DESIGN: Four cases of breast carcinoma containing intraluminal crystalloids were identified among 6900 surgical breast specimens between January 1990 and June 1995 at M. D. Anderson Cancer Center, Houston, Tex. Those sections with crystalloids identified by hematoxylin-eosin stain were stained with periodic acid-Schiff, Alcian blue, and mucicarmine stains. Immunohistochemical and ultrastructural studies and energy-dispersive x-ray analysis were also performed on these sections. RESULTS: The intraluminal crystalloids were eosinophilic, varied in shape and size, and did not exhibit birefringence under polarized light. Immunohistochemically, the crystalloids were negative for keratin, muscle-specific actin, and kappa and lambda light chains, but the surfaces stained positively for epithelial membrane antigen. By electron microscopy, the crystalloids had no limiting membrane and were composed of an electron-dense material with no discernible periodicity. By energy-dispersive x ray element analysis, the crystalloids had no mineral content; however, sulfur was found, indicating a protein content. CONCLUSIONS: The pathogenesis and constituents of these intraluminal crystalloids remain to be determined. Inasmuch as intraluminal crystalloids have not been found in normal ducts or acini, or in ductal hyperplasia without atypia, their presence may serve as a marker for breast carcinoma. PMID- 9199625 TI - Myofibroblastoma of the breast with diverse differentiations. AB - BACKGROUND: The histogenesis of myofibroblastoma of the breast remains unknown. DESIGN: Two cases of myofibroblastoma of the female breast were analyzed using light microscopy and immunohistochemistry. RESULTS: The tumors were well circumscribed and predominantly composed of bland bipolar spindle cells and interspersed bands of hyalinized collagen. Additionally, one tumor contained a cartilaginous island, and the other contained a well-defined small nodule that consisted of fascicular arrangements of mitotically inactive atypical cells, simulating atypical leiomyoma. Both lesions contained a fatty element. Immunohistochemically, spindle tumor cells expressed desmin, alpha-smooth muscle actin, muscle actin, and CD34. The atypical cells were strongly and diffusely positive for all these markers. Both tumors were DNA diploid and had a moderate S phase fraction. The patients have no evidence of disease 4 and 18 months after simple excision. CONCLUSIONS: Myofibroblastoma of the breast may be a benign mesenchymal neoplasm capable of diverse lines of differentiation. PMID- 9199626 TI - Significance of peritoneal washings in gynecologic oncology. The experience with 901 intraoperative washings at an academic medical center. AB - OBJECTIVE: Peritoneal washings are routinely performed during gynecologic surgery. The presence or absence of malignant cells in washings helps determine the stage of the malignancy. However, the efficacy of this procedure has not been studied recently. DESIGN: All intraoperative washings for gynecologic disease at our hospital from 1992 through 1994 (901 cases) were reviewed. Of these, 380 were gynecologic malignancies that were reviewed for changes in staging based on the presence of malignant cells. RESULTS: Histologically, 380 cases were gynecologic malignancies, 521 benign, 79 nongynecologic, and 25 had no accompanying surgical pathology. Of the malignancies, 125 had a diagnosis of cancer on washings. In 12 cases (3.1%), a change in stage resulted. CONCLUSIONS: In a small but significant number of cases, malignant cells in the washings changed postoperative staging, impacting therapeutic measures and prognosis for these patients greatly. Peritoneal washings remain a simple yet effective tool in the evaluation and management of gynecologic malignancies. PMID- 9199627 TI - Pulmonary alveolar microlithiasis. A clinicopathologic and chemical analysis of seven cases. AB - OBJECTIVE: To determine the clinical features and outcome of patients with pulmonary alveolar microlithiasis and to determine the chemical composition of the microliths. CASE MATERIAL: We studied seven cases of pulmonary alveolar microlithiasis. The patients were six women and one man, aged 19 to 70 years (mean age 44.5 years). Clinically, five patients were known to have suffered from this condition for 5 to 41 years. One patient presented with shortness of breath, and another had a gradual decrease in exercise tolerance. None of the patients had a previous history of disturbances in metabolism or any other relevant medical condition. Reports on radiographic studies were available in six cases, and chest radiographs were available for review in the seventh case. They all showed diffuse bilateral pulmonary infiltrates. Open lung biopsies were performed in two patients, and autopsy lung material was reviewed in five patients. RESULTS: Histologically, the lung showed the typical features of pulmonary alveolar microlithiasis, that is, presence of numerous microliths filling the alveolar spaces with either a normal or thickened fibrotic interstitium. Chemical analysis performed on the lung tissue of six of these patients revealed that the microliths consisted principally of calcium and phosphorus salts. Five of these patients died of respiratory failure; however, their deaths occurred from 5 to 41 years following the initial diagnosis. No follow-up information was obtained in two patients. CONCLUSIONS: The findings of this study confirm that pulmonary alveolar microlithiasis can be seen in any age group and that the microliths are composed principally of salts of calcium and phosphorus. Additionally, these cases confirm that the disease typically follows a protracted course. PMID- 9199628 TI - A histopathologic study of localized portal hypertension as a consequence of chronic pancreatitis. AB - OBJECTIVE: To evaluate the mechanism of localized portal hypertension associated with stenosis or obstruction of the splenic vein in chronic alcoholic pancreatitis. DESIGN: Surgical and autopsy specimens from 12 patients with clinically diagnosed chronic alcoholic pancreatitis were examined histopathologically. Autopsy specimens of 10 normal pancreases served as control tissues. RESULTS: In tissues from 11 of the 12 patients with chronic-alcoholic pancreatitis, fibrosis in the pancreatic parenchyma continuously extended to the wall of the splenic vein; organized thrombus formation with recanalization was found in five patients, phlebosclerosis in four, and no changes were found in two. In two of the five patients with organized thrombus formation in the splenic vein, localized portal hypertension had been clinically diagnosed because of splenomegaly and varicose veins in the fundus of the stomach, but it was not accompanied by liver cirrhosis. In the control tissues, fibrosis was not observed in the peripancreatic tissue or the area surrounding the wall of the splenic vein. CONCLUSION: We regard localized portal hypertension due to stenosis or obstruction of the splenic vein as one of the consequences of peripancreatic fibrosis in chronic alcoholic pancreatitis. PMID- 9199629 TI - Rapid brain autopsy. The Joseph and Kathleen Bryan Alzheimer's Disease Research Center experience. AB - OBJECTIVE: To develop a system for retrieving brain tissue within 1 hour after death in an effective and useful manner. DESIGN: Nurse clinicians were employed as study co-ordinators and were available to families 24 hours each day. SETTING: Autopsies were performed at Duke University Medical Center, Durham, NC, from 1985 through 1995. PARTICIPANTS: Neuropathology faculty, fellows, and residents, autopsy technicians; and brain bank staff. RESULTS: Fifty-one rapid autopsies with a postmortem interval of less than 1 hour have been performed. Four of these were normal controls, three were disease controls, and 44 represented Alzheimer's disease patients. Tissue retrieved at rapid autopsy has been distributed to 93 research teams, 30 of these located at Duke University Medical Center. Many researchers have received multiple shipments of tissue. CONCLUSIONS: The Bryan Alzheimer's Disease Research Center Rapid Autopsy Program at Duke University Medical Center has been successful in retrieving tissue from individuals with dementia and also from controls within 1 hour of death. The critical features of the success of this program have been the use of nurse clinicians who work closely with patients and their families to ensure a successful autopsy at the time of death and the maintenance of a 24-hour call schedule for nurses and neuropathology staff. Similar programs can be implemented for experimental work into the pathogenesis of a wide variety of human diseases in which the examination of human tissue is required. PMID- 9199630 TI - Prevalence of myocarditis at autopsy in Turin, Italy. AB - OBJECTIVE: To evaluate the prevalence of myocarditis in a general hospital in Turin, Italy. DESIGN: We retrospectively reviewed 17162 postmortem records from autopsies routinely performed at San Giovanni Battista General Hospital, Turin, between 1965 and 1994. RESULTS: Applying the so-called Dallas criteria, myocarditis was histologically found in 91 cases (0.53%, 95% CI 0.4 to 0.7). The prevalence increased, reaching a peak between 1985 and 1994 (1.2%, 95% CI 0.9 to 1.6). The disease was found more frequently in patients from 20 to 39 years of age, with no difference between males and females. The present data were compared to those of a previous study, performed in 1985 and 1993 to 1994, in which we had prospectively taken into account 605 autopsies (not comprised in the present retrospective study) with standardized myocardial sampling for histological examination: a 5.1% prevalence was found (nearly five times as high as that retrospectively detected in the same period). CONCLUSIONS: If a standardized method of myocardial samples for microscopic examination is not followed, it is possible that myocarditis is overlooked in an unsuspected number of cases. PMID- 9199631 TI - Artifactual signet-ring-like cells in endoscopic biopsy of gastric lymphoma. AB - OBJECTIVE: To describe the morphologic characteristics of neoplastic lymphocytes with shrinkage artifact of cytoplasm secondary to formalin fixation and/or necrosis, which simulate carcinoma signet-ring cells in endoscopic biopsy. DESIGN: Sixty-eight endoscopic biopsies with gastric lymphoma were studied retrospectively. We selected those biopsies in which artifactual signet-ring-like cells were the main histologic feature and were confused with adenocarcinoma. Mucin stains were performed on all specimens. To support their B-cell phenotype, immunohistochemical study with leukocyte common antigen (CD45), pan B-cell marker L26, keratin, and carcinoembryonic antigens were performed. The diagnosis of gastric lymphoma was confirmed in gastrectomy specimens. SETTING: The distinction between poorly differentiated adenocarcinoma and gastric lymphoma in endoscopic biopsies is sometimes difficult owing to the morphologic similarities that these neoplasias can share and the small amount of tissue obtained by this technique. An additional factor that may contribute to this confusion is the presence of artifactual lymphocytes resembling signet-ring cells. RESULTS: Three (4%) of the 68 biopsies showed artifactual lymphocytes in most or all the tissue fragments that resembled carcinoma signet-ring cells. These biopsies showed massive substitution of gastric glands by lymphomatous infiltrate. Crush artifact of neoplastic lymphocytes was observed in many fields. Necrosis, inflammation, and epithelial erosion were prominent findings. Mucin stains were negative, and immunohistochemical studies were positive for leukocyte common antigen and B-cell marker L26 in two of the three cases. CONCLUSIONS: We conclude that if a poorly differentiated neoplasm consistent with signet ring adenocarcinoma is found in an endoscopic biopsy with artifactual changes, the diagnosis of gastric lymphoma must be excluded using histochemical and immunohistochemical studies. PMID- 9199632 TI - Intracranial chondromyxoid fibroma. Report of a case and review of the literature. AB - Chondromyxoid fibroma is an unusual benign tumor of cartilaginous derivation. We describe a rare example of chondromyxoid fibroma of the frontal-sphenoid junction with orbital infiltration in a 35-year-old Hispanic woman who presented with frontal headaches. Gross total excision was performed. The excised mass was composed of neoplastic cells with chondrocytic features within a myxoid matrix. Bony infiltration was present without infiltration of dura mater or brain tissue. The lack of mitotic activity, low cell density, lack of nuclear pleomorphism, and a fused lobular architectural pattern indicated that the lesion was a chondromyxoid fibroma. The lack of hyaline cartilage helped differentiate the lesion from enchondroma. Our case demonstrates the uncommon occurrence of intracranial chondromyxoid fibroma with orbital infiltration. When faced with an intracranial chondrocytic tumor, it is important to distinguish this neoplasm from enchondroma and chondrosarcoma. PMID- 9199633 TI - Primary pulmonary meningioma. Report of a case and review of the literature. AB - Extracranial meningiomas are rare outside the head and neck region. We report a case of primary pulmonary meningioma, initially detected as a radiographic incidental finding, occurring in an asymptomatic 45-year-old woman. Light microscopic examination of both cytologic and histologic preparations was typical of a classical meningioma and included such features as intranuclear pseudoinclusions, psammoma bodies, and cellular whorls. Immunohistochemistry demonstrated tumor cell positivity for vimentin and epithelial membrane antigen, as is characteristic of meningioma. Ultrastructural analysis showed interdigitating cell membranes and desmosomes, with no evidence of basal lamina, neurosecretory granules, or microvilli. On short-term follow-up, the patient is well and has no evidence of a cranial or spinal meningioma. The previous 10 cases reported in the literature had similar characteristics, including a tendency toward occurrence in middle age to older women, asymptomatic presentation, peripheral lung location, and morphologic features. Finally, other conditions in the differential diagnosis and possible histogenesis are discussed. PMID- 9199634 TI - Demonstration of composite nodal B-cell lymphoma and subsequent Hodgkin's disease by flow cytometry and immunohistochemistry. Case report and review of the literature. AB - Hodgkin's disease has rarely been reported to occur subsequent to a previous non Hodgkin's lymphoma, usually of B-cell type and with a 5 to 7-year median interval between diagnoses. Even rarer is the finding of residual non-Hodgkin's lymphoma at the time of Hodgkin's disease diagnosis. Six such cases have been reported, with five of the six representing "discordant" lymphomas and the other one a "composite" lymphoma. Only four of the six cases (all discordant lymphomas) were supported by immunohistochemical studies; flow cytometric immunophenotyping has not been performed in any of the reported cases. We report a nodal composite lymphoma (B-cell non-Hodgkin's lymphoma and Hodgkin's disease, mixed cellularity), supported by flow cytometric immunophenotyping and immunohistochemical studies, occurring in a patient 5 years after a diagnosis of B-cell non-Hodgkin's lymphoma. This report emphasizes the application and usefulness of flow cytometric immunophenotyping and immunohistochemical studies in such cases. PMID- 9199635 TI - Continuing medical education on the World Wide Web (WWW). Interactive pathology case studies on the Internet. AB - OBJECTIVE: To present interactive online continuing medical education (CME) over the World Wide Web as a more efficient alternative to traditional modes of CME delivery. DESIGN: A departmental Web site available to those with access to the Internet. SETTING: A tertiary-care teaching hospital in the United States. RESULTS: Comprehensive case studies have been developed and are complete with images, text, and questions, including explanations for correct and incorrect responses. The images are linked to pertinent text to maximize their educational value. The cases are easily accessible, user friendly, and fully referenced. The system became operational in January 1996, and the first CME certificate was awarded to a participant shortly thereafter. CONCLUSIONS: Continuing medical education over the World Wide Web is an efficient means of delivering CME to the community at large; it allows participating physicians the latitude to obtain convenient CME credit at their leisure, in contrast to the regimented experience of formal CME conferences or symposiums. The interactive format of the CME cases allows the participant to submit immediate comments or criticism to case authors and receive instant feedback on their own performance, features unavailable in comparable educational software packages. The dynamic environment of the World Wide Web lends itself to the production and dissemination of such flexible forms of CME for the physician and will continue to expand in this capacity into the foreseeable future. PMID- 9199636 TI - The College of American Pathologists, 1946-1996. The surveys program. PMID- 9199637 TI - Conscious sedation clinical guideline. Conscious Sedation Working Group, Medical Association of South Africa. AB - OBJECTIVE: The objective of this guideline is to promote safe conscious sedation technique. OUTCOME: (I) The use of safe conscious sedation techniques with endpoints of patient comfort and anxiolysis without loss of protective reflexes; (II) the use of drugs with a wide margin of safety that will protect against unintended oversedation. EVIDENCE: Based on existing consensus statements and some research reports. VALIDATION: Relevant organisations involved in conscious sedation were asked to select a representative to participate in the MASA Conscious Sedation Working Group. All participants represented organisations. The working group met in January 1996 to consider a draft guideline. The document was amended and circulated to working group members, representative organisations and 35 interested persons for endorsement and comment using a modified Delphi technique. All respondents endorsed the guideline (with or without minor corrections.) The national organisations endorsing the guidelines are listed. RECOMMENDATIONS: The use of drugs, equipment and personnel as directed in such a way that enhances the safety of the patient. Pre-sedation screening for at-risk patients. Intra-sedation monitoring, especially the use of a pulse oximeter. Post sedation care and discharge instructions for the patient. PMID- 9199638 TI - Type II diabetes mellitus clinical guidelines at primary health care level. A SEMDSA Consensus Document, 1997, in association with DESSA, ADSA. AB - OBJECTIVE: To develop effective education and management programmes for all aspects of diagnosis, treatment and prevention of complications of type II diabetes mellitus in the primary care setting. OPTIONS: The approach suggested is intended to be adaptable to both optimal and minimal health care settings and to individual patient differences both in the public and the private sectors. OUTCOMES: Few long-term studies are currently available worldwide comparing patient outcome (In terms of macrovascular morbidity and overall mortality) with the various treatment regimens or the clinical and biochemical targets recommended. As complications cannot be totally prevented with our current state of knowledge and therapeutics, attention must still be focused on secondary and tertiary prevention to slow the progression of complications after they have been detected. EVIDENCE: Working groups of diabetologists, epidemiologists, gynaecologists, primary care physicians, nurse specialist educators, dieticians, podiatrists and patient-interest organisations reviewed and discussed the current scientific knowledge and clinical recommendations as reflected by the latest overseas position and consensus statements on the subject. The format and spirit of the document are based on the 'Guidelines for the management of non-insulin dependent diabetes in Africa, 1996' and adapted through local clinical experience to the needs and realities of our country at a primary health care level setting. RECOMMENDATIONS: These include details on the provision of information for early detection, effective management and integration of clinical services for type II diabetes into a multidisciplinary chronic (non-communicable) diseases programme within the existing and evolving national health care system, emphasis on lifestyle modification, appropriate education and medication, the prevention and treatment of complications and timely referrals to a higher level of health care, and the concept of the patient charter of rights and responsibilities in the hope of achieving an optimal level of self-management within the constraints of the disease. VALIDATION: Relevant organisations involved in the education and care of people with diabetes were asked to select representatives to participate in the guidelines working group of the National Diabetes Advisory Board (SEMDSA). Draft documents on 12 topics were produced, amended and widely circulated for comments, criticisms, consensus and endorsement. SPONSORS: The consensus conference and the complication and publication of this document were supported by a generous educational grant from Servier Laboratories (SA). PMID- 9199639 TI - Papillary (chromophil) renal cell carcinoma: histomorphologic characteristics and evaluation of conventional pathologic prognostic parameters in 62 cases. AB - For more than two decades, papillary renal cell carcinoma has been recognized as a possible distinct clinicopathologic subtype of renal cell carcinoma (RCC). However, the histologic criteria for its diagnosis and the clinical outcome are still debated. In an attempt to clarify the diagnostic criteria and resolve issues pertaining to biologic potential, we have evaluated the histologic spectrum of 62 papillary RCCs and assessed significance of conventional pathologic prognostic parameters (Fuhrrman's nuclear grade [NG], pathologic stage [Robson and TNM], tumor size, multifocality, necrosis, and foam cells) and correlated these with outcome. The mean age of patients was 61.8 years (range 22 83), and males were more commonly affected (1.8:1). Grossly, most tumors were well circumscribed, averaged 6.7 cm in size (range 1.8-18), and were predominantly localized to the renal poles (polar vs. mid-renal, 3:1). Multifocality was a prominent feature (24 cases), and in three cases tumors were bilateral. Microscopically, papillary RCCs were predominantly papillary or tubulopapillary, often with a thick fibrous capsule, foam cells, necrosis, hemorrhage, and multifocality. Thirty-five percent of these tumors were low grade (NG I and II) and 65% high grade (NG III and IV). Sixteen of these tumors presented in a higher stage (stages III and IV), and the overall stage correlated with NG (chi 2, p = 0.009). Tumors were further distinguished by cytoplasmic features: eosinophilic (42%), basophilic (34%) and mixed (24%). Eosinophilic tumors were predominantly high grade, and basophilic tumors low grade (chi 2, p = 0.000). A mean follow-up of 57 months showed progression (metastasis, recurrence, or death due to disease) in 21%, whereas 63% were free of disease. Eleven percent died of unrelated causes, and 5% were lost to follow-up. Kaplan-Meier survival analysis showed that both high NG and stage were strongly associated with decreased survival (p = 0.0000 each), as were decreased foam cell (p = 0.0025) and vascular invasion (p = 0.0002). Comparison of 196 reported cases of papillary RCC, including the current series, with reported large series of conventional RCC indicates that papillary RCC usually presents at an early stage, and stage I (Robson) papillary RCC has better 5 year survival rates (87%-100%) than does RCC of the same stage (65-75%). The overall 5 years survival rate for papillary carcinoma (82-90%) was also higher than that of conventional RCC (44-54%). In a Cox proportional hazard regression model, TNM stage appeared to be the only significant variable (p = 0.0000, hazard ratio 10.1) in predicting outcome among papillary RCC. Based on this experience, we conclude that (a) papillary RCC is a malignant tumor, with a tendency to present at a lower stage, but with a distinct potential for progression and aggressive behavior; (b) stratification of these tumors according to cell type, amount of foam cells, presence or absence of vascular invasion, nuclear grade, and pathologic stage provides useful prognostic information; (c) the better 5-year survival rate of papillary RCC (overall and for stage I tumors) compared with that of conventional RCC suggests that it is a tumor with lower malignant potential. Thus, histologic subcategorization of a renal carcinoma as papillary RCC appears to have prognostic implications. PMID- 9199640 TI - Cellular angiofibroma: a benign neoplasm distinct from angiomyofibroblastoma and spindle cell lipoma. AB - Four cases of a distinctive soft-tissue tumor of the vulva are described. They were characterized by occurrence in middle-aged women (39-50 years), small size (< 3 cm), and a usually well-circumscribed margin. The preoperative clinical diagnosis was that of a labial or Bartholin gland cyst in three of the four cases. The microscopic appearance was remarkably consistent and was characterized by a cellular neoplasm composed of uniform, bland, spindled stromal cells, numerous thick-walled and often hyalinized vessels, and a scarce component of mature adipocytes. Mitotic activity was brisk in three cases (up to 11 mitoses per 10 high power fields). The stromal cells were positive for vimentin and negative for CD34, S-100 protein, actin, desmin, and epithelial membrane antigen, suggesting fibroblastic differentiation. Two patients with follow-up showed no evidence of recurrence. The differential diagnosis of this distinctive tumor includes aggressive angiomyxoma, angiomyofibroblastoma, spindle cell lipoma, solitary fibrous tumor, perineurioma, and leiomyoma. The designation of "cellular angiofibroma" is chosen to emphasize the two principal components of this tumor: the cellular spindle cell component and the prominent blood vessels. PMID- 9199641 TI - A clinicopathologic and immunohistochemical comparative study of cutaneous and intramuscular forms of juvenile xanthogranuloma. AB - The clinical, histopathologic, and immunohistochemical features of 18 cases of cutaneous juvenile xanthogranuloma (JXG) and two cases of intramuscular JXG were compared. Clinically, intramuscular JXG differs from cutaneous JXG in that it seems to affect exclusively infants and toddlers and to occur as solitary lesions in skeletal muscles of the trunk rather than in the head and neck region, the most common location of cutaneous JXG. Also, the lesions of intramuscular JXG, unlike those of cutaneous JXG, tend to be composed of a monotonous population of histiocytelike cells, with rare, if any, foamy macrophages or Touton-type multinucleated giant cells. These atypical histopathologic features together with infiltrating borders and the presence of a few mitotic figures are the main reason for the misdiagnosis of intramuscular JXG. Awareness of this atypical presentation should prevent misinterpretation of childhood sarcomas and lymphoproliferative disorders. Immunohistochemically, the cutaneous and intramuscular lesions of JXG exhibit identical profiles, and PG-M1 is the best immunohistochemical marker of JXG. Most likely, JXG is a pathologic histiocytic proliferation of a benign nature that should be treated conservatively. The present study failed to demonstrate any myofibroblastic participation in JXG. Its cause is obscure, and although the hypothesis of viral infection seems attractive considering the clinical picture of the lesion, the present study failed to demonstrate any association between JXG and Epstein-Barr virus, varicella-zoster virus, or cytomegalovirus infection. PMID- 9199642 TI - A comparative study of pure tubular and tubulolobular carcinoma of the breast. AB - Tubular carcinoma is a distinctive subtype of invasive, well-differentiated mammary ductal carcinoma that has a good prognosis when treated by modified radical mastectomy. Little is known about tubulolobular carcinoma. The purpose of this study was to compare the frequency of prognostic factors in pure tubular carcinoma (PTC) and tubulolobular carcinoma (TLC). We studied 90 cases of pure PTC and 17 cases of TLC. The following results were found for PTC: size 0.5 to 1.8 cm (mean, 1.2 cm); multifocality in 18 of 90 cases (20%); axillary lymph-node dissection performed in 51 patients; positive axillary lymph nodes in six of 51 cases (12%); recurrences in one of 90 cases (1%), local. The following results were found for TLC: size 0.6 to 2 cm (mean, 1.3 cm); multifocality in five of 17 cases (29%); axillary lymph-node dissection performed in 14 patients; positive axillary lymph nodes in six of 14 cases (43%); recurrences in two of 17 cases (12%); one local and one systemic, each after mastectomy. The relationship between multifocality and positive axillary lymph nodes was as follows: In PTC, the percentage of positive axillary lymph nodes in multifocal cases was 33%, and in nonmultifocal cases, 7%; in TLC, the percentage of positive axillary lymph nodes in multifocal cases was 60% and in nonmultifocal cases, 33%. In conclusion, multifocality and positive axillary lymph nodes were more frequent in TLC than in PTC. Multifocality appeared to predispose to positive lymph nodes in both PTC and TLC. The distribution of prognostic factors suggest that TLC is a higher-grade lesion than PTC. Long-term follow-up is needed to correlate multifocality with recurrence after breast conserving therapy. PMID- 9199643 TI - Nodular histiocytic/mesothelial hyperplasia: a lesion potentially mistaken for a neoplasm in transbronchial biopsy. AB - This report describes two examples of nodular histiocytic/ mesothelial hyperplasia as seen in transbronchial biopsy that initially led to serious consideration of neuroendocrine neoplasm or meningioma. The biopsies showed nodular collections of cohesive polygonal or round cells with ovoid or deeply grooved nuclei and a moderate amount of finely granular cytoplasm. Nuclear pleomorphism was mild. Immunohistochemical studies showed few cells staining for cytokeratin and the mesothelial marker HBME-1, whereas most cells were decorated by the histiocytic marker PG-M1 (CD68). This lesion appears to be identical to nodular mesothelial hyperplasia as described in hernia sacs and mesothelial/monocytic incidental cardiac excrescences, and we propose modifying the designation to "nodular histiocytic/mesothelial hyperplasia" to take into account the marked predominance of histiocytes over mesothelial cells. The clues to recognition of the true nature of the lesion are clinicopathologic correlation and identification of strips of low cuboidal (mesothelial) cells in the vicinity, and the diagnosis can be further confirmed by immunohistochemical staining. Nodular histiocytic/mesothelial hyperplasia probably results from irritation to the mesothelial lining by various causes leading to focal aggregation of histiocytes within retraction pockets or crevices of the serosal cavity. PMID- 9199644 TI - Breast involvement by extranodal Rosai-Dorfman disease: report of seven cases. AB - Seven cases of breast involvement by extranodal Rosai-Dorfman disease are presented. The patients were women and their ages ranged from 15 to 84 years. Three patients had disease confined to the breast; one had involvement of the breast and ipsilateral axillary lymph nodes, and two had bilateral breast involvement as well as disseminated systemic disease. In all cases the clinical and radiographic presentation of the breast lesion raised the possibility of a malignant tumor. All but one of the lesions were treated by excisional biopsy. Microscopically, the lesions were relatively circumscribed, often multinodular masses, located in the breast stroma, with or without associated involvement of the subcutaneous tissue and dermis. They were composed of sheets of S-100 protein positive large histiocytes displaying lymphocytophagocytosis, scattered in a polymorphous background of mature lymphocytes and plasma cells. The microscopic differential diagnosis includes idiopathic granulomatous mastitis, infective granulomas, Langerhans' cell histiocytosis, Erdheim-Chester disease, fibrous histiocytoma, and malignant melanoma. PMID- 9199645 TI - A reappraisal of hepatic siderosis in patients with end-stage cirrhosis: practical implications for the diagnosis of hemochromatosis. AB - The aim of this study was to describe the histologic pattern of iron distribution in end-stage cirrhosis due to various causes and to test the reliability of the hepatic iron index (equal to hepatic iron concentration divided by age) in excluding or confirming associated hemochromatosis in such a condition. Large slices of the resected livers of 30 patients transplanted for alcoholic and/or viral end-stage cirrhosis were assessed histologically for iron distribution and biochemically for hepatic iron concentration in the least and the most iron overloaded nodules of each case. HLA-A3 was used as the marker for the hemochromatosis gene in the population studied. Intranodular parenchymal siderosis was found in 23 cases (12 spotty, 11 diffuse) with diffuse intrabiliary iron deposits apparent in only two cases. Although in 14 patients the hepatic iron index was significantly high (> 1.9) so as to suggest hemochromatosis, these cases did not correspond to homozygous hemochromatosis with respect to the prevalence of HLA-A3 antigen. End-stage cirrhosis arising from different causes is frequently complicated by parenchymal siderosis that may mimic hemochromatosis, including a hepatic iron index greater than 1.9. The diagnosis of hemochromatosis in patients with end-stage cirrhosis, even those with a hepatic iron index greater than 1.9, should rely mainly on clinical and histologic data. PMID- 9199646 TI - Lesions of the bones of the hands and feet. AB - Neoplasms and their simulators in the bones of the hands and feet include the majority of those found in other skeletal sites, and a disproportionate number of some. We examined the clinical, radiologic, and pathologic features of 240 lesions of the hand and foot bones. Benign tumors and lesions including reactive and reparative conditions comprised 203 cases. The largest single category of neoplasms was that with cartilaginous differentiation, with enchondromas (29 cases) and chondrosarcomas (15 cases) the most common. Noncartilaginous malignant tumors were infrequent and displayed typical radiologic and pathologic features. Florid reactive periostitis, bizarre parosteal osteochondromatous proliferations, and giant cell reparative granulomas made up a larger percentage of lesions in these locations than in other skeletal sites. Lesions of the bones of the hands and feet may frequently be biopsied or treated at hospitals without large orthopedic tumor services. Thus, it is important for the surgical pathologist to be aware of the frequency and characteristics of lesions which may present in these sites. PMID- 9199647 TI - Genotypic alterations in benign and malignant salivary gland tumors: histogenetic and clinical implications. AB - Loss of heterozygosity (LOH) and microsatellite instability (MI) were examined at 24 microsatellite loci in 46 primary benign and malignant salivary gland tumors. Among the 27 benign tumors, 11 (40.7%), manifested microsatellite alterations in at least one locus; of these, five (18.5%) showed LOH and four (14.8%) had microsatellite instability at two or more loci. Four of 11 pleomorphic adenomas (36.4%) had allele loss on the long arm of chromosome 8. Among the 19 malignant neoplasms examined, 10 (52.6%) and one (5.2%) had allele losses and MI, respectively, at multiple loci; three tumors showed MI at only one locus. Frequent LOH was detected at D8S166 (8q11-12), D17S799, and D17S122 (17p-17p11-2) loci, with an incidence of 40%, 37.5%, and 43%, respectively. In general, malignant neoplasms with LOH exhibited aggressive tumor characteristics. Statistically significant correlation's were found between LOH and pathologic classification (chi 2, p = 0.05), higher grade (p = 0.02), DNA aneuploidy (p = 0.005), and a proliferative index of > 6% (p = 0.005) of the malignant tumors. Carcinomas with 17p loci alterations, including two carcinomas expleomorphic adenoma with concurrent 8q LOH, showed more aggressive features. The results suggested that (a) loci on chromosome 8q may harbor a tumor suppressor gene or genes associated with the development or progression of some salivary neoplasms; (b) alterations on the short arm of chromosome 17 may represent an event related to tumor progression; and (c) tumors with LOH at multiple loci have aggressive biologic characteristics. PMID- 9199648 TI - Localized chronic fibrosing vasculitis of the skin: an inflammatory reaction that occurs in settings other than erythema elevatum diutinum and granuloma faciale. AB - Erythema elevatum diutinum (EED) and granuloma faciale (GF) are chronic, localized forms of cutaneous leukocytoclastic vasculitis that result in patterned (storiform or concentric) fibrosis. EED often occurs in systemically ill patients as bilaterally symmetrical plaques, papules, or nodules, often over the dorsa of joints. GF occurs as one or a few plaques on the face. Eosinophils and plasma cells are prominent in GF, whereas neutrophils are plentiful in EED. Rarely, extrafacial lesions accompany facial ones in GF, and there are a few reports of upper respiratory tract masses with GF-like histologic features. We report on eight patients with solitary cutaneous lesions with histologic features similar to those of EED or GF, but whose clinical picture was not that of either disease. One, whose histology resembled GF, had a large multinodular dermal and subcutaneous mass that persisted despite attempted resection. Unusual histologic findings in other cases included storiform fibrosis with dense infiltrates of plasma cells, branching nerve fascicles admixed with EED-like changes, and EED like areas adjacent to zones mimicking a sclerotic fibroma. Chronic fibrosing venulitis can be seen outside the stereotypic settings of GF and EED and is an inflammatory reaction pattern that does not signify a specific diagnosis. Because of transitions between EED or GF-like areas and those of patterned sclerosis with plasma cell-rich infiltrates, we believe that some "inflammatory pseudotumors" of the skin, and perhaps of other sites could be the result of localized vasculitis. PMID- 9199649 TI - Can ischemic colitis be differentiated from C difficile colitis in biopsy specimens? AB - Pseudomembranous colitis is often caused by Clostridium difficile; however, it may also be due to ischemia. To determine if any histologic features could be used to differentiate C difficile from ischemia, 49 biopsies of pseudomembranous colitis (25 from patients with C difficile colitis and 24 from patients with ischemic colitis) were coded, randomized, and evaluated for the presence of numerous variables, including the amount and distribution of mucosal necrosis, lamina propria hyalinization, and atrophic "micro-crypts." Hyalinization of the lamina propria was seen in 19 cases of ischemia but not in C difficile colitis (p < 0.0001). Atrophic-appearing micro-crypts were seen in 18 ischemic cases and 6 C difficile cases (p < 0.0006). Lamina propria hemorrhage, full-thickness mucosal necrosis, and a diffuse microscopic distribution of pseudomembranes were significantly more common in ischemia than C difficile. Endoscopic examination identified pseudomembranes significantly more often with C difficile than ischemia, while the endoscopic appearance of masses or polyps was seen exclusively in cases of ischemia. The presence of a hyalinized lamina propria appeared to be a specific and sensitive marker for ischemia in colon biopsies with pseudomembranes. The presence of atrophic micro-crypts, lamina propria hemorrhage, full-thickness mucosal necrosis, diffuse involvement of all the surface of all biopsies by pseudomembranes, and the endoscopic impression of a localized process, polyp, or mass were also markers of ischemia, while the endoscopic identification of diffuse pseudomembranes favored the diagnosis of C difficile. PMID- 9199650 TI - Follicular basal cell hyperplasia overlying dermatofibroma. AB - Follicular basal cell hyperplasia (FBCH) overlying dermatofibroma represents aborted or impeded pilar differentiation. Historically, this hyperplasia has been misinterpreted as basal cell carcinoma. In a large series of dermatofibroma (258 cases), those that contained primitive or malformed follicular structures over the lesion (59 cases) were compared with those without such elements (199 cases). Statistical analysis of various clinicopathologic features showed that FBCH was significantly associated with younger age, trunk location, hypercellular dermatofibroma, loss of a Grenz zone, clear cell hyperplasia, and seborrheic keratosis-like change. There was an inverse correlation between epidermal atrophy, lichen simplex chronicus-like change, and lower extremity location with FBCH. Histologic features favoring a diagnosis of FBCH over basal cell carcinoma are the focal nature and superficial location of the lesion, lack of cytologic atypia and mitoses, recognizable components of hair follicle differentiation, focal condensation of mesenchymal cells around basal cell proliferation, and the association of epidermal hyperplasia. Our findings suggest that FBCH, clear cell hyperplasia, and seborrheic keratosis-like change all represent an expression of follicular differentiation overlying dermatofibroma. PMID- 9199651 TI - Kaposi's sarcoma-associated herpesvirus and human herpesvirus 8 (KSHV/HHV8) associated lymphoma of the bowel. Report of two cases in HIV-positive men with secondary effusion lymphomas. AB - This report describes two cases of Kaposi's sarcoma-associated herpesvirus/human herpesvirus-8 (KSHV/HHV8)-associated lymphomas that primarily involved the large bowel and that secondarily caused malignant effusions. Involvement of the gastrointestinal tract is of interest because epidemiologic evidence suggests that KSHV/HHV-8 may be transmitted via the fecal-oral route, and KSHV/HHV8 DNA has been detected in rectal samples from HIV-positive patients. This report describes two HIV-positive men who developed primary KSHV/ HHV8-associated lymphomas of the bowel. Despite similar morphology and immunophenotype, these cases differ from most KSHV/HHV8-associated primary effusion lymphomas, which present with malignant effusions in the absence of a solid tumor mass. The spectrum of KSHV/HHV8-associated lymphomas is expanded to include a subset of primary bowel lymphomas in individuals infected with human immunodeficiency virus. PMID- 9199652 TI - The significance of intraluminal crystalloids in benign prostatic glands on needle biopsy. AB - Based on data from autopsy, radical prostatectomy, and cystoprostatectomy specimens, it has been suggested that the finding of intraluminal crystalloids in benign glands on needle biopsy may indicate a concurrent carcinoma; therefore, repeat biopsy is recommended. We studied data from 56 consecutive needle biopsies from the Johns Hopkins Hospital and Dianon Systems in which the diagnosis of intraluminal crystalloids in benign glands was rendered and follow-up data were subsequently obtained. Cases in which crystalloids were present in glands suspicious for cancer, in glands of high-grade prostatic intraepithelial neoplasia, or in adenosis were excluded from the study. Follow-up data included repeat biopsy results and serum prostatic specific antigen levels. Of the 56 men, 31 (55%) had repeat biopsy (two underwent transurethral resection of the prostate [TURP]); the remaining men were either noncompliant or had medical conditions precluding subsequent biopsy. Of the 31 men who underwent repeat biopsies, 23 (74%) had benign diagnoses, one (3%) had high-grade prostatic intraepithelial neoplasia, and seven (23%) had adenocarcinoma. There was no difference in serum prostate-specific antigen values between those with and without cancer on repeat biopsy. In a control population of men with a benign first biopsy not showing crystalloids, the incidence of cancer on repeat biopsy was 16.2%, which was not statistically significantly different from the incidence found in our study group. We conclude that men with prostate biopsy results showing benign glands with crystalloids are at no significantly higher risk of having cancer on repeat biopsy than if crystalloids were not present. PMID- 9199653 TI - A nodal gamma/delta T-cell lymphoma with an association of Epstein-Barr virus. AB - The postthymic gamma/delta T-cell lymphoma is rare, and most occur as extranodal tumors, e.g., in hepatosplenic or cutaneous forms. We here report an unusual nodal case that initially presented as a T-zone lymphoma. The neoplasm recurred as systemic lymphadenopathy 25 months after complete remission with terminal high grade transformation. Phenotypic analysis showed CD1-, CD2+, CD3+, CD4-, CD5-, CD7+, CD8+, CD10-, CD16-, CD19-, CD20-, CD21-, CD25-, CD56-, CD57-, T-cell receptor (TCR) alpha/beta antigens negative, TCR gamma/delta antigens positive, and an HLA-DR+ phenotype. Cytogenetic studies showed clonal chromosomal translocations involving chromosomes 1, 5, 6, 8, 15, and X in eight of 15 cells; t(X;5;1)(q13;q13;p22) and t(6;15;8)(p22;q26;q13). Genotypic analysis showed the same clone, characterized by the TCR gamma-chain gene rearrangement pattern, to be present in both initial and recurrent tumors. The lymphoma cells were also demonstrated to express the latent membrane protein-1 by immunohistochemistry and EBV-encoded small RNAs by in situ hybridization. Southern blot analysis using the probe of the terminal repeat demonstrated incorporation of multiple copies of EBV in the recurrent tumor. However, the initial lesion, which contained a smaller number of EBV-positive cells, showed no such evidence of clonal proliferation. These data suggest that EBV may be associated with high-grade transformation, although its exact role in lymphomagenesis remains uncertain. The present study also adds to our understanding of the clinicopathologic spectrum of gamma/delta T cell neoplasia. PMID- 9199654 TI - Clear cell dermatofibroma. PMID- 9199655 TI - Papillary carcinoma of the thyroid with stromal lymphoid infiltrate. PMID- 9199656 TI - Marginal zone lymphoma of skin. PMID- 9199657 TI - Inhibition of Ras prenylation: a signaling target for novel anti-cancer drug design. AB - The cancer-causing activity of Ras requires the prenylation of a cysteine fourth from its carboxyl terminus. Rational design of peptidomimetics of the carboxyl terminal tetrapeptide prenylation site on Ras resulted in pharmacological agents capable of inhibiting Ras processing, selectively antagonizing oncogenic signaling and suppressing human tumor growth in mouse models without side effects. This mini-review describes the efforts of several groups to design, synthesize and evaluate the biological activities of farnesyltransferase and geranylgeranyltransferase I inhibitors. Among the important issues that will be discussed are the mechanism of action of these inhibitors and the potential mechanisms of resistance to inhibition of K-Ras farnesylation. PMID- 9199658 TI - Sequence specificity, enantiospecificity and polyelectrolyte effect in the binding to DNA of a 6-(2-pyridyl)phenanthridine chiral photonuclease. AB - In order to establish the basis for the rational design of a novel family of intercalating chiral photonuclease drugs aimed at photochemotherapy, namely N, N' dialkylated 6-(2-pyridinium)phenanthridinium (pyp) dications, a detailed investigation of the DNA binding of the dq2pyp member (where dq2 stands for CH2CH2-), was conducted. The study addresses the sequence- and enantiospecificity, as well as polyelectrolyte effects in the drug-DNA interaction. Binding isotherms with synthetic polynucleotides, forcefield calculations, affinity chromatography in a DNA-cellulose stationary phase and salt-dependent equilibrium and kinetic studies with DNA were used. dq2pyp shows a strong preference for alternating GC over AT base pairs; binding to homopolymeric DNA is weak (< 3 x 10(4) M-1). Affinity chromatography shows enantiospecific binding of dq2pyp to DNA. The polyelectrolyte contribution to the binding free energy are shown to be relatively important (-4.8 kcal/nmol out of an overall value of -7.2 kcal/mmol at 10.2 mM Na+). The slope of the logkd (dissociation rate constant) vs. log[Na+] plot (0.7) agrees with the values predicted from counterion condensation theory for a dicationic intercalator, giving further support to such a DNA binding mode for dq2pyp. The relatively high kinetic dissociation constants (logkd = 0.70log[Na+] + 3.79) in comparison with those of propidium (two orders of magnitude larger at any Na+ concentration) seems to originate from the absence of amino groups in dq2pyp. The kinetic association constants (logka = -1.06log[Na+] + 5.53) are twice these of propidium, probably due to the less restrictive positioning of dq2pyp at the intercalation site. The kinetic studies support a mechanism of intercalation in which the drug forms a pre-equilibrium outside the complex followed by the intercalation of the drug. Molecular modelling is used throughout to rationalize all the experimental data, as well as to propose new candidates with improved DNA affinity and residence time. PMID- 9199659 TI - Effects of Dex-Verapamil on Doxorubicin cytotoxicity in P388 murine leukemia cells. AB - Resistance-modifying agents (RMAs) such as Verapamil have been proved to be effective in reversing multi-drug resistance (MDR) in many in vitro assays. In this study we have investigated the efficacy of Dex-Verapamil, the R-isomer of Verapamil, as a chemosensitizer in a murine leukemia cell line (P388) and in its resistant counterpart (P388/Dx) expressing a typical MDR phenotype. We have examined in vivo the effect of the co-administration of Dex-Verapamil and Doxorubicin in mice transplanted with P388 or P388/Dx cells. Mice treated with the combination of Doxorubicin plus RMA had a significant increase in survival rate as compared to controls; however, the effect was modest. On the contrary, in vitro Dex-Verapamil can enhance Doxorubicin cytotoxicity in P388/Dx cells with a much greater effect depending on the treatment scheme used, by increasing the intracellular content of drug. Taken together our data indicate that Dex Verapamil can indeed increase the sensitivity to Doxorubicin in resistant cells, but the limited efficacy shown in vivo demonstrates that this phenomenon is strongly dependent on the treatment scheme used and on the maintenance of constantly elevated serum levels. PMID- 9199660 TI - Synthesis, DNA binding and cytotoxicity of 1-[[omega-(9 acridinyl)amino]alkyl]carbonyl -3-(chloromethyl)-6-hydroxyindolines, a new class of DNA-targeted alkylating agents. AB - We report the first synthesis of examples of the seco-CI DNA alkylating moiety 3 (chloromethyl)-6-hydroxyindoline linked to a 9-aminoacridine DNA-intercalating unit (compounds 1a-1c). The sequence-specificity of DNA alkylation by these compounds was studied by the gel electrophoresis cleavage assay. In contrast to the known trimethoxyindole-linked compound (1d), which alkylates exclusively at N3 of adenines in the minor groove, the acridine-linked analogues (1a-1c) alkylate predominantly at the N7 of guanines in the major groove (the first CI analogues reported to do so), although DNase I footprinting experiments show that the initial non-covalent binding of 1a-1c is not base sequence selective. DNA unwinding experiments show that the acridine moiety of 1a-1c remains intercalated after alkylation. PMID- 9199661 TI - The effect of antitumor trans-[PtCl2(E-iminoether)2] on B-->Z transition in DNA. AB - The B-->Z transition of DNA modified by platinum(II) complexes has attracted considerable interest because of a possible relationship with the molecular mechanism of antitumor activity of these metal-based compounds. Until recently it was generally accepted that the cis geometry of leaving groups in the bifunctional platinum(II) complexes should be therapeutically active. This paradigm had to be abandoned recently due to the finding that several analogues of clinically ineffective trans-diamminedichloroplatinum(II) (transplatin) exhibit antitumor activity. One of these therapeutically active trans compounds is trans-dichlorobisiminoetherplatinum(II) (trans-EE) [iminoether = E-HN = C(OMe)Me]. In view of the fact that DNA is the main pharmacological target for platinum(II) complexes and that these complexes attack preferentially guanine residues in DNA, it is of interest to examine the effect of iminoether platinum(II) complexes on the conformation of double-stranded poly(dG-dC) and its synthetic analogues. As these polymers can undergo the B-->Z transition, we have investigated in detail the effect of trans-EE and its cis isomer (cis-EE) on this conformational transformation using different techniques. They include circular dichroism spectroscopy, Raman spectroscopy and immunochemical assay employing specific antibodies against Z-DNA. It has been shown that cis-EE somewhat facilitates the B-->Z transition induced by increasing NaCl concentration and radically lowers its cooperativity. The polymers modified by cis-EE at low or high salt concentrations have been found to adopt distorted conformations which belong to the B and Z families respectively. Thus, cis-EE affects the B-->Z transition in DNA in a way similar to antitumor cis-diamminedichloroplatinum(II) (cisplatin). On the other hand, trans-EE was found to affect B-->Z transition (its cooperativity or the NaCl concentration corresponding to the midpoint of the salt-induced transition) only slightly. This behavior of trans-EE was, however, fundamentally different from that of clinically ineffective transplatin, which hinders B-->Z transition and lowers its cooperativity. The different effects of trans-EE on the B-->Z transition in comparison with the effects of cisplatin, transplatin and monofunctional chlorodiethylenetriamineplatinum(II) chloride are consistent with a unique DNA binding mode of this new antitumor trans compound, which might be associated with its unexpected biological efficacy. PMID- 9199662 TI - Immunohistochemical localization of the Na-K-Cl co-transporter (NKCC1) in the gerbil inner ear. AB - We mapped the cellular and subcellular distribution of the Na-K-Cl co-transporter (NKCC) in the adult gerbil inner ear by immunostaining with a monoclonal antibody (MAb T4) generated against human colon NKCC. Heavy immunolabeling was seen in the basolateral plasma membrane of marginal cells in the stria vascularis and dark cells in the vestibular system. Subpopulations of fibrocytes in the cochlear spiral ligament and limbus and underlying the vestibular neurosensory epithelium also stained with moderate to strong intensity, apparently along their entire plasmalemma. Because MAb T4 recognizes both the basolateral secretory (NKCC1) and the apical absorptive (NKCC2) isoforms of the co-transporter, we employed reverse transcription and the polymerase chain reaction (RT-PCR) to explore isoform diversity in inner ear tissues. Using NKCC1 and NKCC2 isoform-specific PCR primers based on mouse and human sequences, only transcripts for NKCC1 were detected in the gerbil inner ear. The presence of abundant NKCC1 in the basolateral plasmalemma of strial marginal and vestibular dark cells confirms conclusions drawn from pharmacological and physiological data. The co-expression of NKCC1 and Na,K-ATPase in highly specialized subpopulations of cochlear and vestibular fibrocytes provides further evidence for their role in recycling K+ leaked or effluxed through hair cells into perilymph back to endolymph, as postulated in current models of inner ear ion homeostasis. PMID- 9199663 TI - Use of enzyme overlay membranes to survey proteinase activity in frozen sections: cathepsin-like and plasmin-like activity in regenerating newt limbs. AB - We present a method that permits extremely simple and rapid screening of proteolytic enzyme activity in sectioned tissues. Enzyme overlay membranes (EOMs) are custom-made membranes designed to fluoresce at sites of specific proteolytic enzyme activity after separation of proteins by gel electrophoresis. EOMs, selected to detect either plasmin-like or cathepsin B-like activity, have been used in a novel way to document the distribution of enzyme activity in frozen sectioned tissues. When moistened membranes were placed in contact with sectioned regenerating newt limbs, a fluorescent pattern of enzyme activity was generated. In limbs at 3 hr post amputation, cathepsin B-like activity was prominent across the amputation site but plasmin-like activity was distributed in dermal and deeper proximal tissues, suggesting different roles for these two classes of enzymes. EOM enzymology in situ (EEI) on frozen sectioned tissues may be a widely useful technique to display distribution and level of activity of proteolytic enzymes in various systems. PMID- 9199664 TI - Differential expression of c-fos in vitro by all anterior pituitary cell types during the estrous cycle: enhanced expression by luteinizing hormone but not by follicle-stimulating hormone cells. AB - C-fos expression appears in some activated cell types. Because of dynamic changes in gonadotropes during the estrous cycle, this study was initiated to determine if fos might be expressed in gonadotropes before any period of activation. We detected c-fos and pituitary antigens in dissociated anterior pituitary cells by dual-labeling immunocytochemistry. The highest percentage of cells with fos protein were found in proestrous rat populations. In diestrous and proestrous populations, dual labeling showed that 6-9% of pituitary cells contained fos with adrenocorticotropin, thyroid-stimulating hormone, prolactin, or growth hormone antigens. In contrast, only 0.8-3% contained fos with luteinizing hormone (LH) or follicle-stimulating hormone (FSH) antigens. We then tested the hypothesis that gonadotropes might increase fos expression earlier in the cycle. In populations from metestrous rats, c-fos labeling was found in 45% of LH cells compared to only 23% of LH cells in the proestrous group. This suggests that proportionately more LH cells are being activated to produce fos early in the cycle. Perhaps fos is used in translation of LH beta antigens or gonadotropin-releasing hormone (GnRH) receptor mRNAs. In contrast, less than 1% of all pituitary cells expressed fos with FSH at all stages of the cycle (only 6-12% of FSH cells). This differential expression suggests one mechanism behind the regulation of non parallel storage and release of gonadotropin antigens. PMID- 9199665 TI - Prohormone convertases in mouse submandibular gland: co-localization of furin and nerve growth factor. AB - Nerve growth factor (NGF) in mouse submandibular glands (SGs) is generated from a 35-kD precursor by proteolytic enzymes that have yet to be identified. Prohormone convertases (PCs) cleave the NGF precursor in vitro, and in this study we questioned whether PCs could process salivary NGF in vivo. mRNA coding for PC2 (but not PC1) was detected on Northern blots of SG mRNA and also by in situ hybridization within parasympathetic neurons of intralobular ganglia. Northern blot and in situ hybridization analyses also detect mRNA coding for furin. In SGs of male mice, furin mRNA levels are high at birth and remain high throughout development. In glands from female mice, levels decline during postnatal development and are lower in adults than in newborns. Immunocytochemistry detects furin immunoreactivity in pro-acinar and ductal cells of glands from newborn and pubescent mice. In glands of adults, furin immunoreactivity is detectable in acinar cells but highest levels are present in NGF-containing granular convoluted tubule cells. These data, taken together with those from previous studies, suggest that furin is a candidate processing enzyme for NGF in mouse submandibular glands. PMID- 9199666 TI - Modulation of protein kinases and microtubule-associated proteins and changes in ultrastructure in female rat pituitary cells: effects of estrogen and bromocriptine. AB - This study focused on the intracellular signal transduction system and microtubule-associated proteins (MAPs), such as MAP-2 and Tau protein. The modulation of these proteins and their correlation with ultrastructural changes were investigated in rat pituitary prolactin (PRL) cells. Adult female Wistar rats were treated with estrogen and bromocriptine and their pituitary glands were removed for analysis of the expression of tubulin, MAP-2, Tau protein, protein kinase C (PKC), and calcium calmodulin (CaM) kinase. Western blot analysis showed that estrogen increased and bromocriptine decreased the expression of PKC alpha, beta 1, beta 2, CaM kinase alpha, beta, MAP-2, and Tau protein. MAP-2 and Tau protein, which are cytosolic proteins, being translated on free ribosomes, were associated with the membrane of whirling rough endoplasmic reticulum (RER) in estrogen-treated cells and dissociated with vesiculated RER induced by bromocriptine. These results suggested that the modulation of MAP-2 and Tau protein may reflect changes of PKC and CaM kinase, and that the quantitative changes and intracellular modulation of MAPs induced by estrogen and bromocriptine, i.e., estrogen-induced association and bromocriptine-induced dissociation of MAP-2 and Tau protein with membrane of RER, may reflect the dynamics of microtubules and are associated with structural changes in the RER and changes in the synthesis and intracellular transport of PRL. PMID- 9199667 TI - Complex co-localization of chromogranins and neurohormones in the human gastrointestinal tract. AB - Co-localization of chromogranin (Cg) A, B, and C has been studied in different neuroendocrine cell types in histologically normal mucosa from human gastrointestinal tract (corpus, antrum, duodenum, ileum, and colon) using single , double-, and triple-immunofluorescence stainings. Virtually all enterochromaffin (EC) cells contained CgA, and those in the luminal two thirds of the antral mucosa and villi of small intestine often also contained CgB. A few EC cells in the duodenal crypts contained CgC. Most gastrin cells harbored both CgB and CgA, although rather more CgB than CgA, but some gastrin cells contained all three types, i.e., also CgC. Some CCK cells also contained all three chromogranins. Enteroglucagon cells in the duodenal villi contained CgA and some CgB. CgA (but not B or C) was found in some secretin, GIP, enteroglucagon/peptide YY, and neurotensin cells. A few somatostatin cells contained CgA but neither CgB nor CgC. CgA and C were found mainly in the basal cell region, whereas CgB occurred more diffusely throughout the cytoplasm. This varying distribution suggests that not all secretory granules contain CgA, or that CgB may occur in a nongranular form. The varying composition of the different chromogranins may reflect their complex functional roles in the widespread neuroendocrine system. PMID- 9199668 TI - Cloning and expression of murine SC1, a gene product homologous to SPARC. AB - A number of cDNAs (SC1, QR1, and hevin) have been shown to be similar to SPARC (secreted protein acidic and rich in cysteine), a matricellular protein that regulates cell adhesion, cell cycle, and matrix assembly and remodeling. These proteins are 61-65% identical in the final 200 residues of their C-termini; their N-terminal sequences are related but more divergent. All have an overall acidic pl, with a follistatin-like region that is rich in cysteine, and a Ca+2 binding consensus sequence at the C-terminus. Using degenerate primers representing the most highly conserved region in SPARC, SC1, and QR1, we identified a 300-BP SC1 clone in a primary polymerase chain reaction (PCR) screen of a mouse brain cDNA library. This cDNA was used to obtain a full-length clone, which hybridized to a 2.8-KB RNA abundant in brain. Mouse SC1 displays a similarity of 70% to mouse SPARC at the amino acid level. Northern blot and RNAse protection assays revealed a 2.8-KB mRNA expressed at moderate levels (relative to brain) in mouse heart, adrenal gland, epididymis, and lung, and at low levels in kidney, eye, liver, spleen, submandibular gland, and testis. In contrast to SPARC, in situ hybridization showed expression of SC1 mRNA in the tunica media and/or adventitia of medium and large vessels; transcripts were not detected in capillaries, venules, or large lymphatics. The distribution of transcripts for SC1 was also different from that of SPARC in several organs, including adrenal gland, lung, heart, liver, and spleen. Moreover, SC1 mRNA was not evident in endothelium cultured from rat heart, bovine fetal and adult aorta, mouse aorta, human omentum, and bovine retina. Cultured smooth muscle cells and fibroblasts also failed to express SC1 mRNA. The absence of SC1 transcript in cultured cells indicates that the SC1 gene is potentially sensitive to regulatory factors in serum or to a three-dimensional architecture conferred by the extracellular matrix that is lacking in vitro. In conclusion, the expression of SPARC and SC1 appears to be coincident in specific tissues (e.g., adrenal gland and brain), but these proteins exhibit distinct expression patterns in most organs of the mouse. Because SC1 and SPARC are structurally similar and exhibit counteradhesive effects on cultured cells, their overlapping and/or adjacent expression in most tissues predicts that one protein might compensate functionally, at least in part, for the other. PMID- 9199669 TI - Phenotypic modulation of smooth muscle cells after arterial injury is associated with changes in the distribution of laminin and fibronectin. AB - Earlier in vitro studies suggest opposing roles of laminin and fibronectin in regulation of differentiated properties of vascular smooth muscle cells. To find out if this may also be the case in vivo, we used immunoelectron microscopy to study the distribution of these proteins during formation of intimal thickening after arterial injury. In parallel, cell structure and content of smooth muscle alpha-actin was analyzed. The results indicate that the cells in the normal media are in a contractile phenotype with abundant alpha-actin filaments and an incomplete basement membrane. Within 1 week after endothelial denudation, most cells in the innermost layer of the media convert into a synthetic phenotype, as judged by loss of actin filaments, construction of a large secretory apparatus, and destruction of the basement membrane. Some of these cells migrate through fenestrae in the internal elastic lamina and invade a fibronectin-rich network deposited on its luminal surface. Within another few weeks a thick neointima forms, newly produced matrix components replace the stands of fibronectin, and a basement membrane reappears. Simultaneously, the cells resume a contractile phenotype, recognized by disappearance of secretory organelles and restoration of alpha-actin filaments. These findings support the notion that laminin and other basement membrane components promote the expression of a differentiated smooth muscle phenotype, whereas fibronectin stimulates the cells to adopt a proliferative and secretory phenotype. PMID- 9199670 TI - Inducible nitric oxide synthase in the anterior pituitary gland: induction by interferon-gamma in a subpopulation of folliculostellate cells and in an unidentifiable population of non-hormone-secreting cells. AB - In the context of immune-endocrine relationships, we have previously shown that interferon-gamma (IFN-gamma) inhibits hormone secretion in anterior pituitary (AP) cell cultures. The non-hormone-secreting folliculostellate (FS) cells were found to mediate this inhibitory action. Because in the immune system IFN-gamma is a strong stimulator of nitric oxide (NO) release through the induction of NO synthase (NOS), we investigated whether the inducible form of NOS (iNOS) is present in (rat) AP cell cultures, and whether its expression is stimulated by IFN-gamma. Immunocytochemistry revealed that under basal in vitro conditions only a very few AP cells contained iNOS. Treatment with IFN-gamma caused a sixfold rise in the number of iNOS-positive cells and augmented the intensity of the staining. The increased number of iNOS-expressing cells was paralleled by elevated production of NO. Some of the iNOS-positive cells extended cytoplasmic processes between hormone-secreting cells, which is a characteristic of FS cells. Immunostaining of FS cell-poor and FS cell-enriched populations (obtained by gradient sedimentation) also suggested the presence of iNOS in a subpopulation of FS cells. By double immunofluorescence techniques we found that about 65% of iNOS expressing cells were positive for S-100, a marker protein for FS cells. However, around 80% of the S-100-positive cells were not labeled for iNOS. On the other hand, the majority of the S-100-negative iNOS-containing cells could not be further identified by antisera against the classical AP hormones, suggesting the presence of iNOS in a still unidentified non-hormone-secreting cell type of the AP gland. This report is the first to demonstrate the expression of the inducible form of NOS in the AP gland. IFN-gamma upregulates this expression, showing that cytokines may use the same signalling mechanisms in both the immune and the endocrine system. In addition, a putative new function of a subpopulation of FS cells in the paracrine regulation of the AP gland is suggested. PMID- 9199671 TI - Immunohistochemical similarities and differences between amelogenin and tuftelin gene products during tooth development. AB - Amelogenins and tuftelins are highly specialized proteins secreted into the developing enamel matrix during mammalian enamel formation. Both tuftelins and amelogenins have been associated with various functions during nucleation and maturation of the developing enamel matrix. In this study we conducted experiments to investigate whether tuftelins and portions of the amelogenin molecule were deposited and processed in spatially distinguished portions of the developing enamel matrix, using antibodies specific against tuftelin or amelogenins. The amelogenin antibodies were raised against recombinant and native amelogenins and also included an antibody against a polypeptide encoded by amelogenin exon 4. To compare spatial expression patterns of enamel protein epitopes, 3-day postnatal mouse molar tooth organs were processed for paraffin histology and cut into serial sections. Adjacent sections were exposed to antibodies against either tuftelin or various amelogenin epitopes. To investigate age-related changes of enamel protein expression, amelogenin and tuftelin antibodies were applied to tooth organs of developmental stages E19 and 1, 3, 5, 7, 9 and 11 postnatal days. Tuftelin was detected within the odontoblast processes during earlier stages of development (E19 and 1 day postnatal), whereas during later stages (3-11 days) it was recognized in a portion of the enamel layer adjacent to the dentine-enamel junction. In contrast, all four antibodies against amelogenins reacted with parts of the ameloblast cytoplasm and the entire enamel layer. Using immunohistochemistry, we were not able to detect any differences in the spatial distribution of the four amelogenin epitopes investigated. The spatial differences in the distribution of amelogenin and tuftelin as observed in this study may be interpreted as an indication of functional differences between both proteins during early enamel biomineralization. PMID- 9199672 TI - The epidermal stem cell compartment: variation in expression levels of E-cadherin and catenins within the basal layer of human epidermis. AB - The basal layer of the epidermis contains two types of proliferating keratinocyte: stem cells, with high proliferative potential, and transit amplifying cells, which are destined to undergo terminal differentiation after a few rounds of division. It has been shown previously that two- to three-fold differences in the average staining intensity of fluorescein-conjugated antibodies to beta 1 integrin subunits reflect profound differences in the proliferative potential of keratinocytes, with integrin-bright populations being enriched for stem cells. In the search for additional stem cell markers, we have stained sections of normal human epidermis with antibodies to proteins involved in intercellular adhesion and quantitated the fluorescence of individual cell cell borders. In the basal layer, patches of brightly labeled cells were detected with antibodies to E-cadherin, beta-catenin, and gamma-catenin, but not with antibodies to P-cadherin, alpha-catenin, or with pan-desmocollin and pan desmoglein antibodies. In the body sites examined, palm and foreskin, integrin bright regions were strongly labeled for gamma-catenin and weakly labeled for E cadherin and beta-catenin. Our data suggest that there are gradients of both cell cell and cell-extracellular matrix adhesiveness within the epidermal basal layer and that the levels of E-cadherin and of beta- and gamma-catenin may provide markers for the stem cell compartment, stem cells expressing relatively higher levels of gamma-catenin and lower levels of E-cadherin and beta-catenin than other basal keratinocytes. PMID- 9199673 TI - Immunohistochemical localization of arginine and citrulline in rat renal tissue. AB - Using antibodies highly specific for L-arginine and L-citrulline, we localized these amino acids in rat kidney with immunohistochemical methods. Highest levels of arginine immunoreactivity were observed in epithelial cells of proximal tubules in the outer stripe of the outer medulla and the collecting ducts in the cortex. Staining intensity of proximal convoluted tubules in the outer stripe decreased from the inner side to the outer side. In the inner medulla, collecting ducts were labeled with moderate intensity. Staining within the cortex was apparent only with collecting ducts. Citrulline immunoreactivity was localized in the epithelial cells of collecting ducts both in the cortex and medulla. Immunoreactivity was also found in glomerular podocytes and in the epithelial cells of proximal convoluted tubules in the outer medulla. These localizations were different from those of other amino acids previously reported. The precise cellular distribution of arginine and citrulline in rat kidney was determined for the first time by an immunohistochemical method in the present study. PMID- 9199674 TI - PDGFR alpha expression during mouse embryogenesis: immunolocalization analyzed by whole-mount immunohistostaining using the monoclonal anti-mouse PDGFR alpha antibody APA5. AB - We investigated the cells that express platelet-derived growth factor receptor alpha (PDGFR alpha) during mouse embryogenesis. PDGFR alpha expression has been identified by in situ hybridization or immunohistochemistry using polyclonal antibodies on tissue sectins. Because no immunostaining study using whole-mount specimens has been published to date, we established a new monoclonal antibody (MAb), APA5, for this purpose. Our results differed in that APA5 stained only the paraxial mesoderm, whereas other investigators concluded that most if not all mesodermal cells expressed PDGFR alpha. Moreover, we did not find PDGFR alpha expression in embryonic erythrocytes, which have been previously suggested to express PDGFR alpha. On the basis of our present results, we wish to revise the proposed PDGFR alpha expression as follows. At the pregastrulation stage, PDGFR alpha is expressed only in primitive endoderm, particularly that in the ectoplacental cone. On gastrulation, it is expressed at high levels in the paraxial mesoderm. This expression continues after its differentiation into the somite. Along with the differentiation and migration of the sclerotome, PDGFR alpha + cells begin to become distributed throughout the embryonal mesenchyme. During organogenesis, particularly intense staining is detected in regions of epithelial and mesenchymal interaction, such as the tooth bud and bronchi. In addition to mesodermal derivatives, the developing lens, apical ectodermal ridge, glial precursor, cardiac valves, and choroid plexus express PDGFR alpha. Our results with whole-mount immunostaining show that PDGFR alpha is abundantly expressed and may play important roles during embryogenesis. PMID- 9199675 TI - A simple enzyme histochemical method for the simultaneous demonstration of acetylcholinesterase and monoamine oxidase in fixed-frozen sections. AB - We describe an enzyme histochemical technique for the simultaneous demonstration of acetylcholinesterase (AChE) and monoamine oxidase (MAO) (Types A, B, or A+B) in fixed-frozen sections. Several regions in the mesencephalon and brainstem were examined for both somatic and neuropil labeling. The results obtained are equivalent or superior to those obtained using previous methods for the individual localization of these enzymes. The simultaneous visualization of AChE and MAO in the same section allows the relationship of the two enzymes to be easily assessed with brightfield microscopy. PMID- 9199676 TI - Management of arteriovenous malformations: Part II. PMID- 9199677 TI - Gamma knife radiosurgery. PMID- 9199678 TI - An analysis of the natural history of cavernous malformations. AB - BACKGROUND: The treatment of cavernous malformations has been controversial. Some reports suggest that surgical resection of the lesion for the prevention of recurrent hemorrhage should not be considered because of low hemorrhagic risk. However, the role of surgery in management of cavernous malformations is undergoing reevaluation. The decision for surgical resection should be based on a careful analysis of the natural history of this lesion, which is not well understood. METHODS: We investigated, retrospectively, the natural history of 108 cavernous malformations in 62 patients. Individual cavernous malformations were divided into four categories on the basis of magnetic resonance (MR) findings. The pattern of clinical and radiologic presentation and outcomes of management were analyzed. RESULTS: The age of the patients ranged from 4-63 years (mean: 32.2 years). Multiple lesions were found in 13 of 62 patients (21%) and two of these patients were siblings. Twenty-five out of 62 patients had suffered recurrent symptoms. The bleeding rate was 2.3%/person/year (1.4%/lesion/year) during 2509.6 patient years. There were no significant differences between the bleeding rates of each type of lesion. During the follow-up period of 12-48 months (mean: 22.4 months), two of 28 patients conservatively treated had recurrent hemorrhages (rebleeding rate: 3.8%/person/year). During the follow-up period of 12-66 months (mean: 21.7 months), recurrent hemorrhages were observed in two of 17 patients with radiosurgery (rebleeding rate: 7.8%/person/year). CONCLUSION: Our study has provided a profile of the natural history of these lesions. Based on our results, we recommend surgical excision of cavernous malformations in those patients with recurrent symptoms or acute progressive symptoms. PMID- 9199679 TI - Development and prevention of frozen shoulder after acute aneurysm surgery. AB - BACKGROUND: We conducted a study on periarthritis humeroscapularis or "frozen shoulder," a postoperative complication of aneurysm surgery. The purpose of this study was to seek the cause of this complication and the methods of preventing it in patients who undergo aneurysm surgery. METHODS: The diagnosis of frozen shoulder was based on the clinical presence of shoulder pain and difficulty in raising arms that developed within 3 months of surgery. Sixty-four patients who underwent aneurysm surgery with no motor deficit were examined and classified into three groups: (1) early surgery (29 patients in the acute stage after subarachnoid hemorrhage); (2) delayed surgery (19 patients in the chronic stage); and (3) elective surgery (16 patients with unruptured aneurysms). RESULTS: The incidence of frozen shoulder was 41% in the early surgery group, 16% in the delayed surgery group, and 13% in the elective surgery group. The highest incidence of frozen shoulder was found to occur in the early surgery group and was attributed to the immobility of their upper extremities during postoperative treatment. Since patients who undergo surgery in the acute stage are often delirious and confused for several days after surgery, their arms are tied down by their sides in order to prevent them from inadvertently removing catheters such as the one for ventricular drainage. It seems that this manner of immobilizing the patient's arms is the cause of the development of frozen shoulder: Our study showed that if each arm was passively raised by turns above the patient's shoulder, the patient was able to maintain the range of motion of the upper arms and was less likely to develop frozen shoulder. CONCLUSION: Inactivity of the shoulder joints due to immobilizing the upper extremities of patients after acute aneurysm surgery seemed to cause the development of frozen shoulder. The incidence of this complication was greatly reduced by keeping the patient's upper arms raised alternately to maintain their range of motion after acute aneurysm surgery. PMID- 9199680 TI - MRI artifacts following anterior cervical diskectomy. AB - BACKGROUND: Magnetic resonance imaging (MRI), despite being an excellent imaging technique in neurosurgical practice, is unfortunately susceptible to numerous artifacts. Some of these artifacts are easily identifiable and do not interfere; however, others are more subtle and can be easily mistaken for false pathology. Postoperative MRI can further complicate the imaging interpretation, by producing another group of artifacts. It is imperative for practicing neurosurgeons, as well as neuroradiologists, to have a clear understanding of these postoperative artifacts. METHODS: We discuss four cases who had been operated for anterior cervical decompression with bony fusion. All the patients had a postoperative MRI of the cervical region that showed a "false compression" of the cervical cord. The normal computed tomography (CT) scan in some cases and the discrepancy with the clinical condition of the patients excluded the diagnosis of compression of the cervical cord. RESULTS: The overall appearance of the postoperative MRI can be very difficult to interpret. The artifact seen following anterior cervical diskectomy is an example of such a situation. We have confirmed that the postoperative MRIs showing artifacts do not indicate cord or root compression; a routine postoperative plain X ray or CT scan of the operated area can also confirm the absence of compression. CONCLUSION: These are examples of cases in which the postoperative MRI had an unexpected metallic artifact that not only caused difficulty in the interpretation of the images but at times suggested a clinical problem when actually there was none. Very thin cut CT scans may not show these artifacts that are picked up by the sensitive MRI study. A proper clinical evaluation and selection of the appropriate MRI techniques and the MRIs can eliminate or at least decrease the incidence of the artifacts. Above all, further education of practicing physicians is needed to avoid false alarms caused by these metallic artifacts. PMID- 9199681 TI - Surgical treatment of solid brain stem tumors in adults: a report of 22 cases. AB - BACKGROUND: Brain stem tumors in adults are infrequent. Most reports of surgical treatment for these tumors involve partial tumor removal in highly selected patients. A more aggressive approach for removing tumors, especially solid and intrinsic ones, has been controversial. METHODS: Twenty-two adult patients with brain stem tumors were surgically treated. Surgical techniques, potential risks, and selection of appropriate treatment were evaluated. RESULTS: Tumors were totally or subtotally removed in 20 patients and only partially removed in two patients. Serious complications such as respiratory disturbances and circulatory dysfunction occurred in 10 patients. Eight patients with these complications recovered after appropriate treatments. Upon discharge, most signs and symptoms improved in 17 patients. CONCLUSION: Most brain stem tumors, except for malignant gliomas and small ventral tumors, are amenable to an aggressive surgical approach. Exophytic medullary tumors that present dorsally comprise the most benign subgroup of brain stem tumors. Total removal can enhance survival, improve the patient's quality of life, and offer a favorable long-term prognosis. Appropriate management of postoperative complications is essential for good results. PMID- 9199682 TI - Transsphenoidal adenomectomy in Cushing's disease via a lateral rhinotomy approach. AB - BACKGROUND: Cushing's disease may be treated by surgical pituitary adenomectomy. We present a surgical approach to the pituitary gland that increases the possibilities of a selective adenomectomy, and compare our results with those of other studies. METHODS: A retrospective study of patients with Cushing's disease undergoing transsphenoidal selective adenomectomy via a lateral rhinotomy at Sahlgrenska University Hospital from 1984-93 is presented. Thirty-one patients (26 women, five men; mean age: 44 years, range: 13-75 years) with Cushing's disease were followed for a median time of 4.5 years after operation (range: 1-10 years). Preoperative and postoperative urinary and serum cortisol, and circadian rhythm of serum cortisol were measured. We also measured serum TSH, T4, PRL, FSH, LH, and testosterone as well as urine and plasma osmolality. RESULTS: Our remission rate was 77% and the recurrence rate 3%. Hormonal insufficiency was rare. Hypothyroidism and hypogonadism were present in 3% of the patients, and diabetes insipidus occurred in 6% of the patients. CONCLUSION: Selective adenomectomy with its good opportunities for cure and improvement should be regarded as the treatment of choice for Cushing's disease. Using the lateral rhinotomy approach to the sphenoidal cavity results in good accessibility to the sella turcica and its pituitary adenomas, a low frequency of postoperative pituitary insufficiency, and a high remission rate. PMID- 9199683 TI - Chordoma: a case report. AB - BACKGROUND: Chordomas are tumors of notochordal origin that account for approximately 1%-4% of all primary malignant bone tumors. The majority of patients with chordomas have a poor surgical prognosis due to extent of disease at diagnosis. These lesions have been previously classified based solely on their location. METHODS: We describe here a case report of a posterior epidural C5-T1 chordoma that was discovered in a young patient who presented with weakness and paresthesia in all four extremities. This lesion was notable for its extraosseous and extradural characteristics. RESULTS: C5-T1 laminectomy with gross total resection of the mass led to complete resolution of all symptoms. There has been no evidence of tumor recurrence to date. CONCLUSIONS: We propose here a new classification system for chordomas that emphasizes the difference in resectability of these lesions depending on the space they occupy and the presence or absence of an osseous connection. PMID- 9199684 TI - Cerebellar pilocytic astrocytoma with leptomeningeal dissemination: case report. PMID- 9199685 TI - Multiple infected extradural parasellar hydatid cysts. AB - BACKGROUND: Intracranial hydatid disease constitutes 1%-2% of all cases of hydatid disease. Multiple, infected, extradural, parasellar hydatid cysts in a patient constitutes an extremely rare presentation. CASE REPORT: This 21-year-old man presented with a progressive left supraclavicular swelling of 3 years duration and raised intracranial pressure of 6 months duration with a past history of left-sided chronic suppurative otitis media that had resolved with antibiotics. On neurologic examination, he had bilateral deterioration of vision with optic atrophy; right temporal field defect; left III, IV, VI, VI, and V2 cranial nerves palsy; and left ear conductive deafness. The patient's E.S.R was raised. His computed tomography (CT) scan showed a hypodense, lobulated lesion in the middle cranial fossa with a hypodense, nonenhancing rim, septations, and focal calcification without perifocal edema. A purulent fluid was aspirated from the left supraclavicular swelling, which did not reveal any organism on staining and culture. Aspiration of the left temporal swelling showed whitish watery fluid, the cytology of which revealed an infected hydatid cyst. Excision of the left temporal extradural, hydatid cysts was done, except the portion of the capsule adherent to the dura, and albendazole was started. One month later, the supraclavicular hydatid cysts were removed. Six months later, a left mastoidectomy was performed for chronic suppurative otitis media. A repeat CT scan showed complete resolution of the hydatid cysts. There was no recurrence at 1 year follow-up. CONCLUSIONS: A rare case of multiple infected extradural hydatid cysts of the parasellar region is reported. The unusual CT picture of a hypodense lobulated mass with septations and a hyperdense rims is presented. The difficulties in its complete excision and successful management with long-term albendazole therapy are discussed. PMID- 9199686 TI - Hypertrophic cranial pachymeningitis involving the pituitary gland: a case report. AB - BACKGROUND: As the use of magnetic resonance imaging (MRI) has become widespread, the reported cases of pachymeningitis have increased. Various inflammations are known to occur in the pituitary gland. However, an association between pachymeningitis and pituitary inflammation has not been recognized previously. CASE DESCRIPTION: This 56-year-old woman complained of incessant headache and deafness. MRI showed thickened dura at the right convexity and a dumbbell shaped pituitary enlargement, which was isointense to the gray matter (T1-weighted images) and homogeneously enhanced. Microscopic examination of the dura mater showed a marked increase of collagen fibers and various inflammatory cells. The anterior pituitary lobe was infiltrated with lymphocytes, and normal tissues were displaced by increased collagen fibers. CONCLUSION: Hypertrophic cranial pachymeningitis, a rare clinical entity, is a progressive lesion prone to include the dura mater at the skull base and can be fatal. We report a case of hypertrophic cranial pachymeningitis involving the pituitary gland, and discuss the possible etiology of the association. PMID- 9199687 TI - A case of giant cell reparative granuloma of the petrous bone: demonstration of the proliferative component. AB - BACKGROUND: Giant cell reparative granuloma (GCRG) is an uncommon benign lesion of the bone. It typically arises in the mandible and rarely involves the skull. The cytologic nature and genesis of the involved cells are poorly understood. METHODS AND RESULTS: We report a case of GCRG in the petrous bone of a 3-year-old girl. One year following gross total removal, the granuloma recurred locally and was resected en bloc at the second surgery. Histologically, the lesion was composed of oval or spindle-shaped stroma cells admixed with a number of multinucleated giant cells. Immunohistochemical study demonstrated that 5.6% of the stroma cells, but none of the multinucleated giant cells, were positive for MIB-1 antibody. CONCLUSION: These results suggest that this lesion expands by proliferation of the stromal component, with a growth rate roughly between those of the typical benign and malignant brain tumors. The cytologic nature of the cells comprising this uncommon disease remains to be elucidated. PMID- 9199688 TI - MRSA meningitis and intrathecal injection of arbekacin. PMID- 9199689 TI - Inadvertent intraoperative myelography with Hypaque: case report and discussion. AB - BACKGROUND: Myelography is routinely performed safely using nonionic water soluble radiographic contrast media. However, inadvertent introduction of ionic contrast media into the thecal space can result in a syndrome of spasms and convulsions, which can lead to death if not recognized and dealt with in a timely manner. METHODS: We report a case of inadvertent use of the ionic diatrizoate meglumine, an ionic contrast agent, instead of a nonionic contrast agent during intraoperative myelography. RESULTS: The patient developed a sterotypical syndrome of ascending myoclonic spasms, resulting in rhabdomyolysis. Treatment included elevation of the head, removal of cerebrospinal fluid, administration of anticonvulsants, diuresis and sedation, and neuromuscular blockade. The patient recovered well, and there were no long-term sequelae. CONCLUSIONS: Intrathecal introduction of ionic contrast media and the resultant syndrome must be recognized promptly and treated with aggressive medical management to address rhabdomyolysis and seizures. Ionic contrast media should be stored and marked in such a way as to avoid inadvertent use in myelography. PMID- 9199690 TI - Invasive monitoring of limbic epilepsy using stereotactic depth and subdural strip electrodes: surgical technique. AB - BACKGROUND: In spite of advances in noninvasive localization of seizure foci, some cases of intractable limbic epilepsy still require invasive recordings in order to identify the site of seizure onset. This necessitates a safe and reliable method for placing depth and subdural electrodes in the mesial temporal and orbitofrontal regions. The University of Florida has devised a system that utilizes CRW-based stereotactic placement of bitemporal depth electrodes in conjunction with placement of subdural strips over the inferolateral temporal lobe and orbitofrontal cortex. This report describes the surgical technique and initial clinical experience using this method. METHODS: Depth electrodes are placed along the long axis of the hippocampi via an occipital approach. A CRW based stereotactic system was developed that incorporates both computed tomography (CT) and magnetic resonance imaging (MRI) for selection of target and entry sites and displaying the electrode trajectory. Subdural strip electrodes are placed over the inferolateral temporal and orbitofrontal regions through burr holes. RESULTS: This method has been used in 18 patients (depth electrodes only in three patients and depth electrodes with subdural strips in 15 patients). This information lead to surgical resections in 15 patients. No resection was recommended in three patients (two with bitemporal onset and one with no seizures after 6 weeks). Complications were limited to an unplanned removal of one electrode and an asymptomatic lateral temporal lobe contusion in one patient. CONCLUSIONS: This method provides a safe and effective way to sample bilateral mesial temporal and orbitofrontal regions in cases of intractable limbic epilepsy. PMID- 9199691 TI - Brain stem compression secondary to adipose graft prolapse after transpetrosal approach: case report. AB - A case of fat graft prolapse into the prepontine cistern with clinical deterioration is presented. The patient had undergone a transpetrosal approach for a very large craniopharyngioma and had autologous thigh fat strips to obliterate the mastoid cavity; postoperatively she suffered an important deterioration from adipose graft prolapse. Clinical presentation, MRI findings, treatment, and avoidance of this complication are discussed. PMID- 9199692 TI - The blood supply of the hypoglossal nerve: the microsurgical anatomy of its cisternal segment. AB - BACKGROUND: While the characteristics of the vasculature of the second (intracanalicular) segment of the hypoglossal nerve are well known, the vascularization of the first (cisternal) segment of this nerve has not been examined so far. Many pathologic processes and malformations can be located in the premedullary cistern, which may affect the vasculature of the cisternal segment. Consequently, we decided to examine the blood supply of the cisternal segment. METHODS: The anatomic features of the cisternal segment and its vasculature were examined in 15 hypoglossal nerves after injection of india ink and gelatin into the vertebrobasilar arterial system. RESULTS: The cisternal segment was noted to consist of 3-15 long roots, which usually formed two trunks of the hypoglossal nerve. The roots of each nerve received blood from the anterolateral and the lateral medullary arteries, which ranged from 3 to 5 in number and between 100 microns and 500 microns in caliber. These arteries may arise from the perforating branches or the pontomedullary branch of the basilar artery; the vertebral artery or its perforators; the anterior spinal artery or its vascular roots; the posterior spinal artery; and the posterior inferior cerebellar artery. The main hypoglossal arteries, which ranged in diameter from 20 microns to 80 microns, always coursed along the dorsal surface of the roots of the hypoglossal nerve. CONCLUSIONS: The cisternal segment of the hypoglossal nerve was always vascularized by several vessels, which mainly originated from the vertebral artery and its branches. This observation was discussed from the neurosurgical point of view. PMID- 9199693 TI - The Canadian Health Care System. PMID- 9199694 TI - Capillary telangiectasia of the pons. PMID- 9199695 TI - The future of neurovascular surgery. Part I: Intracranial aneurysms. PMID- 9199696 TI - Effects of diethyl-dithiocarbamate on antioxidant system in carp tissue. AB - The influence the pro- and antioxidant values the diethyl-dithiocarbamate (DDTC) in different concentrations in carp tissues was studied. From antioxidant enzyme superoxide dismutase, catalase and glutathion peroxidase activity changes were studied in tissue homogenates. Beside enzyme activities tissue lipid peroxidation (LP), reduced glutathione (GSH) and hydroxyl radical loading of the tissues were measured. Our results indicate that DDTC affects antioxidant parameters by generating GSH excess, not loading the cells thereby with "oxidative stress". PMID- 9199697 TI - Unconditional discrimination as a paradigm for investigating visual processing in the avian brainstem. AB - Visual discrimination between colors and patterns was studied in telencephalectomized chicks of Japanese quail lines that were artificially selected for early approach preferences for particular colors and patterns. Since these preferences are not conditional on prior learning, they provide an effective experimental paradigm to study visual discrimination without reliance on acquired stimulus discrimination. The data indicate that the quail's brainstem thalamus complex is sufficient for relatively fine discriminations between various colors and achromatic patterns. PMID- 9199698 TI - The morphology of the intestine of the entomophagous longfingered bat, Miniopterus inflatus: mucosal topography and possible landmarks. AB - The intestinal tract of the longfingered bat, Miniopterus inflatus, was studied macroscopically, with the light microscope and the scanning electron microscope. The intestine comprised a small mass of coiled loops contained in a rather small abdominal cavity. Macroscopically, the stomach was of the simple type and the intestine was a short convoluted tube whose diameter decreased craniocaudally. A caecum, an appendix and a colon were absent and the only portion of the large intestine observed was a short rectum grossly identifiable only on the account of its greater diameter. Microscopically, a small initial part of the intestine bordering the pylorus was characterized by numerous pits of variable sizes and shapes. This segment preceeded the ridge-like, transversely oriented villi that occupied the rest of the foregut. These villi were tallest in the proximal parts of the foregut and decreased in height caudally, ceasing completely at the junction between the small intestine and the rectum. Goblet cells were few in the cranial part of the intestine and increased caudally, reaching a maximum in the rectum. Intestinal glands were abundant in the region between the villi but Brunner's glands were absent in the submucosa. Generally the intestine of Miniopterus resembles that of the other bats which have been studied but showed structural details suggestive of an increased digestive and absorptive efficiency. PMID- 9199699 TI - Morphometry of the kidneys of fresh water percoid and ostariophysian teleosts. AB - Kidneys of four fresh water teleost species representing percormorpha (Oreochromis niloticus and Micropterus salmoides) and Ostariophysi (Cyprinus carpio and Clarias mossambicus) were studied for quantitative structural characteristics using tissues fixed by perfusion. The renosomatic index (weight of kidney/weight of fish) was higher in the ostariophysian than percoid fish (p < 0.05) but values of volume of nephronic tissue/body weight were not significantly different. Results obtained by point counting morphometry showed that the kidneys of the percoid species were made up of 56.85-64.4% nephronic tissue, 30.45-32.47% blood vessels, 2.84-8.20% ureter and ureteral ducts and 2.07-2.48% connective tissue. The kidneys of the osariophysian species comprised of 35.27-36.71% nephronic tissue, 27.86-29.29% blood vessels, 1.59-4.20% ureter and its ducts, 29.07-29.51% interstitial cells and 2.70-3.60% connective tissue. The mean values for the volume proportions of the components of nephronic tissue were not significantly different in the percoids and ostariophysians and they were; renal corpuscles 3.84-6.54%, neck segments 0.50-1.11%, proximal segments 67.17-69.72%, distal segments 18.16-20.55%, and collecting tubule-collecting duct (CT-CD) system 5.66-7.41%. These results show that the quantitative structural characteristics of the kidneys of fresh water species from different orders are quite similar and emphasise the all persuasive influence of the environment on renal structure and function in teleosts. PMID- 9199700 TI - Phenotypic heterogeneity of the progeny of Streptomyces griseus conidia. AB - In order to understand the complex ontogenetical processes, the development of Streptomyces (S.) griseus was applied as a model. The developmental cycle of S. griseus starts and ends as a conidium. In between, coenocytic mycelium develops which, if studied by cytomorphological or biochemical methods, exhibits conspicious heterogeneity. The hyphae develop into young, transient and old vegetative hyphae and different stages of reproductive forms. In developmentally blocked mutants these sequences of events appear mixed in all possible associations. It seems as if the program of development could be divided into several subprograms. The quantitative evaluation of the results show that the individual morphological markers exhibit certain independence from each other realized with a given probability. The conidia of S. griseus are also heterogeneous concerning all morphological and physiological traits examined so far (shape, size, light refraction, staining and shape of nucleoids with Feulgen, methyl green--pyronine, intensity and form of polysaccharide distribution, heat resistance, etc.). Kinetics of the survival curves of two S. griseus strains--a well-sporulating and its developmentally blocked mutant /24/--are different from each other, one has many more heat resistant conidia than the other but the kinetics of the survival curves of the two S. griseus strains indicate that spore populations of both react differently to heat treatment and heat resistance can be modeled by assuming the presence of two independent subpopulations of spores with different heat sensitivity. The emergence of two distinct subpopulations with (possibly) the same genetic make-up is designated: phenotypic segregation. Heat resistance is first of all species specific (genetically determined) but the epigenetic segregation seems to be characteristic of the developmental process. This process can in certain mutants be affected by environmental conditions and more importantly by the so-called autoregulators (A-factor and factor C). Factor C and A-factor are needed to normal development, if their quantity or the time of addition to the culture was not optimal, the quantity of spores decreased. PMID- 9199701 TI - Genetic analysis of the components of winterhardiness in barley (Hordeum vulgar L.). AB - Winterhardiness in cereals is the consequence of a number of complex and interacting components: cold tolerance, vernalization requirement and photoperiod sensitivity. An understanding of the genetic basis of these component traits should allow for more effective selection. Genome map-based analyses hold considerable promise for dissecting complex phenotypes. A 74-point linkage map was developed from one hundred double haploid lines derived from a winter x spring barley cross and used as the basis for quantitative trait locus (QTL) analyses to determine the chromosome location of genes controlling components of winterhardiness. Despite the greater genome coverage provided by the current map, a previously-reported interval on chromosome 7 remains the only region where significant QTL effects for winter survival were detected in this population. QTLs for heading date under 24 h light map to the same region. A QTL for heading date under this photoperiod regime also maps to chromosome 2. A distinct set of QTLs mapping to chromosomes 1, 2, 3 and 5 determined heading date under 8 h light. Patterns of differential QTL expression underscore the complexity of Winterhardiness. PMID- 9199702 TI - Comparative studies on potato tuber development using an in vitro tuber induction system. AB - A method for synchronized in vitro tuber induction in a Hungarian cultivar of Solanum tuberosum designated "Keszthelyi 855" has been developed. It was shown that in this system tuberization and stolon elongation primarily depend on the level of sucrose in the medium. The cytokinin, 6-bensylaminopurine (BAP), also enhances the efficiency of tuber formation, however, only at sucrose concentration above 4% (w/v). The synchronized plant culture provided starting material for isolation of genes specifically expressed in tuberizing Solanum species during the early stage of tuber development. In comparison with the non tuberizing Solanum brevidens, three types of specific transcripts have been obtained by differential screening. Based on DNA sequence analysis the genes isolated code for the major tuber proteins, patatin and proteinase inhibitors. PMID- 9199703 TI - Study of the alcohol dehydrogenase-1 (Adh1) gene in tetraploid corn: expression in the pollen grains and restriction fragment length polymorphism. AB - The results of two separate experiment are presented in this paper. In the first experiment pollen assays were carried out in Wf9 tetraploid maize plants in order to decide whether the lack of the ADH enzyme activity in the pollen grains was caused by a 'O' allele or an active transposon. On the basis of the results we suppose that the reduced ADH enzyme activity in the pollen grains is the result of a transposon affecting only the gametophyte. In the second experiment a maize Adh1-S genomic clone was constructed by polymerase chain reaction (PCR) and used as a probe to detect polymoprhism around the Adh1 gene in W117 S18 tetraploid maize sublines. The PCR20-EcoRI clone-enzyme combination resulted in a monomorphic RFLP pattern. The PCR20-Bgl II probe-enzyme combination yielded a multiple-banded pattern. PMID- 9199705 TI - Utilisation of population genetic and multivariate methods in the evaluation of results of cattle type traits judgement. AB - The results of the evaluation of linear type traits evaluation of Hungarian beef cattle populations were analysed by means of both traditional population genetic and multivariate biometric methods, i.e. by means of cluster analysis based on factor analysis. By using the multivariate methods it was possible to evaluate the different types of cattle with respect to type traits, reproductivity and productivity properties. It is proven by the results obtained that factor analysis is practicable in the judgement at the population level of the individual properties. A great number of type traits--27 or more--making an interrelating system can be analysed well by this method whilst the cluster analysis based on the factor analysis is suitable to classify the population on the basis of individual evaluation with simultaneous consideration of all type traits. PMID- 9199704 TI - Production of fish chimeras from embryonic cells. AB - The efficiency of two cell-transplantation methods were compared for the production of embryonic cell derived fish chimeras. The classic microinjection technique (blastula stage donor cells are microinjected into blastula embryos) was compared to a novel aggregation method for fish developed by our group. This method utilises the ability of dechorionated fish embryos to aggregate. Morula cells dissociated in Ca2+, Mg2+ free medium were aggregated to recipient embryos of different developmental stage. Donor cells and recipient embryos of different developmental stages were used for most efficient incorporation in the chimeras. THe fate of donor cells derived from blastulae, was followed by labelling them with FITC-dextrane (FD). The most efficient transplantations were gained by using 16-32 cell stage recipients for aggregation (18% survival of chimeras at swim up stage). Labelled donor cells were contributing to the embryos in varying ratio. A comparison of the efficiency of aggregation was made between diploid-diploid, diploid-haploid and diploid-interspecfic (diploid) hybrid chimeras. In all three cases chimeras containing different proportion of donor cells were gained. After one day incubation the embryos were dissociated by trypsin digestion and number of labelled and non-labelled cells were counted under fluorescent microscope. Experiments were performed on Rosy barb (Barbus conchonius), Carp (Cyprinus carpio) and African catfish (Clarias gariepinus). PMID- 9199706 TI - Changes in alkylation damage removal during in vitro neuronal differentiation. AB - Mouse teratocarcinoma cell lines allow the analysis of very early commitment and differentiation events, that are likely to be similar to those operating in the early mouse embryo. We have previously characterised the excision repair capabilities of these cells after ultraviolet light irradiation and found that differentiation is accompanied by reduction of excision repair. In the present study we examine the operation of an other DNA repair pathway participating in the removal of alkylation damage. O6-alkylguanine-DNA alkyltransferase (ATase) activity was determined in undifferentiated and differentiated mouse P19 teratocarcinoma cell line. To obtain more information about regulation of ATase we transfected P19 cells with constructs harbouring a human ATase cDNA driven by a housekeeping promoter. PMID- 9199707 TI - Molecular genetic studies in monogenic and polygenic human diseases. AB - The main goal of this study was to determine and characterise the types of mutations in two monogenic human disorders: cystic fibrosis (CF) and Duchenne/Becker muscular dystrophy (DMD, BMD) and the susceptibility allele frequency in a polygenic disease: type I insulin-dependent diabetes mellitus (IDDM). After analysing 220 chromosomes for mutations in the CF (Cystic Fibrosis Transmembrane Conductance Regulator = CFTR) gene, delta F508 mutation was most abundant (41%) and out of the non-delta F508 CF mutations 5% was identified as G542X, G551D, R553X, N1303K and W1282X. The CF haplotype analysis by using linked markers to the CFTR gene revealed that the CF "B" haplotype occurred in 66.7% of patients, and this haplotype was 57.2% in patients carrying the delta F508 mutation. Prenatal genetic diagnosis for CF was performed in 10 fetuses: 3 were affected, 6 were carriers, and 1 without any CF mutation. Fifty % of 66 patients with DMB/BMD muscular dystrophy had one or more exon deletions in the dystrophin gene. Eighty-five % of the deletions occurred at the 3' and 15% at the 5' end of the gene. Out of the three prenatal diagnosis in one case DMD was substantiated. Thirty-six % of 50 patients with IDDM possessed four, 44% three and 20% two susceptibility markers in the HLA-DQA1, -DQB1 region. The onset of the disease correlated with the number of susceptibility alleles. PMID- 9199708 TI - Monoclonal antibodies to the distinct antigenic sites on glycoproteins C and B and their protective abilities in herpes simplex virus infection. AB - The relative importance of the humoral immune response to various antigenic sites on the glycoproteins C and B (gC, gB) of herpes simplex virus (HSV) was evaluated in BALB/c and DBA/2 mice passively immunized with monoclonal antibodies (MoAbs) and then challenged with lethal dose of infectious virus. Eight MoAbs to three topographically distinct antigenic sites on gC and eight MoAbs to two distinct antigenic sites on gB were selected. The results indicated that any antigenic site on gC and gB contains epitopes for the protective immunity. However, individual MoAbs to different epitopes of the same antigenic site (i.e. antigenic site III on gC, and antigenic site II on gB) varied extremely in their protective ability. The protection did not correlate with the virus neutralization in vitro whereas it correlated significantly with the immune cytolysis in the presence of complement. The information about protective epitopes is essential for understanding the immunology of HSV infection at molecular level and may have implications for the design of HSV vaccine. PMID- 9199709 TI - Biological characteristics of an enzootic subtype of western equine encephalomyelitis virus from Argentina. AB - In order to expand our knowledge on the biological characteristics of an enzootic South American subtype of western equine encephalomyelitis (WEE) virus, strain AG80-646, we inoculated guinea pigs, rabbits, newborn chickens and Vero and chick embryo cell cultures with this and other WEE and Wee-related viruses. AG80-646 was found apathogenic for guinea pigs even when inoculated intracranially (i.e.) or intraperitoneally (i.p.), and the animals did not develop viraemia. AG80-646 killed rabbits and the animals developed high viraemia (peak titer was 7.0 log PFU/0.1 ml). These data and previous serological evidence led us to look for a mammal as a natural host. AG80-646 was found lethal for newborn chickens inoculated subcutaneously (s.c.) (peak viraemia titer was 6.6 log PFU/0.1 ml). AG80-646 produced plaques (diameter 0.8-1.0 mm) in Vero and chick embryo cells 3 4 days post infection (p.i.) A comparison of AG80-646 with other WEE complex virus strains led to the following observations: (1) The lethality for guinea pigs was high for the two epizootic Argentinian strains, Cba 87 and Cba CIV 180, zero for the two enzootic strains, AG80-646 and BeAr 10315 (virus Aura), and intermediate for the Russian strain Y62-33 (low by i.c. route and zero by i.p. route); (2) AG80-646 was more virulent for rabbits inoculated i.p. than the three epizootic strains Cba 87, Cba CIV 180 and McMillan; (3) AG80-646 was less virulent for new-born chickens than the Argentinian epizootic strain Cba CIV 180; (4) The viraemia level correlated always with the strain virulence in each animal host. This study provides tools for the differentiation of WEE complex viruses and strains in the future ecological work on WEE in South America. PMID- 9199710 TI - Contribution to the regulation of virus replication in cells latently infected with human immunodeficiency virus 1. AB - Monocytes/macrophages have been known to play an important role in the initiation and propagation of human immunodeficiency virus 1 (HIV-1) infection. To analyze the function of these cells during the clinical asymptomatic period of infection, we examined the effect of murine peritoneal macrophages and human peripheral blood macrophages on two cell lines latently infected with HIV-1, a promonocytic cell line, U1, and a T-cell line, ACH-2. Monokines of the murine peritoneal macrophages induced significant viral expression in U1, but not in ACH-2 cells. Experiments employing transient transfection of U937 and CEM cells with HIV long terminal repeat (LTR)-chloramphenicol acetyl transferase (CAT) plasmids indicated that the effect of these monokines was due to specific activation of the HIV LTR. In contrast, supernatants of human macrophages induced viral expression in both ACH-2 and U1 cells. These results suggest that several monokines are active in regulating the transition from the clinical asymptomatic period of HIV infection to progression to acquired immunodeficiency syndrome (AIDS). PMID- 9199711 TI - Clinical value of specific intrathecal production of antibodies. AB - The production of intrathecal antibodies is considered a highly specific marker for an infection of the central nervous system (CNS), e.g. borreliosis or tick borne encephalitis (TBE). To investigate the validity of this assumption, we examined records of patients who had been hospitalized between 1989 and 1995, who were tested for borreliosis (n = 8003) and TBE (n = 904) and whose cerebrospinal fluid (CSF) had subsequently tested positive for intrathecal production of antibodies. The time period between the beginning of the symptoms and the time of the CSF examination ranged from one day to six weeks. Seventy-seven patients showed a production of intrathecal antibodies against Borrelia burgdorferi. Three of these patients were false positives with no history and no clinical signs of neuroborreliosis. In two cases, this was due to a non-specific cross-reaction caused by a preceding infection with syphilis. The third false positive was possibly caused by an earlier administration of immunoglobulins. Three patients showed a production of intrathecal antibodies against TBE virus. Two of these patients were false positives. In one case, we suspect that the production of intrathecal antibodies was caused by a non-specific immune reaction during an acute neuroborreliosis. One year earlier, the patient had contact with TBE virus through a vaccination against TBE. The cause of the second false positive is unclear, the clinical findings, acute encephalitis and the serological analysis suggest a cross-reaction with a virus similar to TBE. A specific intrathecal production of antibodies is not a proof for an infection of the CNS. In unclear cases, one should carry out a Western blot analysis or, if one suspects a case of TBE, a neutralization test. PMID- 9199712 TI - Serological relationships between tobacco necrosis virus-inducible cucumber peroxidase and corresponding peroxidase isoenzymes in other Cucurbitaceae. AB - Pathogenesis-related peroxidases (PRXs), found in leaves of different cucurbits infected with tobacco necrosis virus (TNV), were compared serologically using a highly specific rabbit antiserum raised against PRX purified from TNV-infected cotyledons of Cucumis sativus L., cv. Laura. After native polyacrylamide gel electrophoresis (PAGE) of intercellular washing fluid extracts from a large number of species of this family, immunoblot analysis revealed the presence of cross-reacting protein bands in all species tested. Despite the serological relatedness, the analysis also revealed that the antiserum recognized only fast moving PRX isoenzymes localized in the apoplast space. PMID- 9199713 TI - Identification and partial characterization of beta-1,3-glucanase from virus infected cucumber cotyledons. AB - One of the five "pathogenesis-related" (PR) proteins known to accumulate in Cucumis sativus L. cv. Laura and to react hypersensitively to tobacco necrosis virus (TNV) was shown to to have beta-1,3-glucanase activity. The TNV-induced acidic beta-1,3-glucanase activity increased 6-fold after infection and had extracellular localization and estimated M(r) of 25,700. The beta-1,3-glucanase activity was investigated with a new method of activity staining using a conjugated substrate in overlay gels. By using the antiserum against tobacco beta 1,3-glucanase purified to homogeneity, a close serological relationship was demonstrated between cucumber and tobacco beta-1,3-glucanases on immunoblots. PMID- 9199714 TI - Sequence analysis and comparison of 190 K surface antigen gene fragment of a new species of spotted fever group rickettsiae. AB - A 533 bp long PCR product amplified from rickettsial strain HL-93 DNA with the primer pair Rr 190.70p and Rr 190.602n, designed from DNA sequence encoding 190 K protein antigen of R. rickettsii, was cloned into plasmid vector PGEM-T and sequenced. The primer-flanking region of the product, an open reading frame, was 491 bp long. The sequence of the product was compared with those of the corresponding regions of DNAs of R. rickettsii (strain R), R. japonica (strain VR 1363) and R. conorii (strain Malish 7) which were reported earlier by other authors. The results showed that 23, 31 and 52 nucleotides in the compared sequence in strain HL-93 differed from those in R. japonica, R. rickettsii and R conorii, respectively. The homologies of strain HL-93 with R. japonica, R. rickettsii and R. conorii were 95.6%, 94% and 90% in nucleotide, and 89%, 87% and 80% in putative amino acid sequences. We consider strain HL-93 as a new member of the spotted fever group (SFG) rickettsiae on the basis of a high degree of homology and genetic divergence in the nucleotide sequence of a part of the 190 K protein gene. PMID- 9199715 TI - Optimization of purification procedure for potato virus Y strain NN. AB - Potato virus Y strain NN (PVYNN) was purified from mechanically infected plants Nicotiana tabaccum cv. Samsun by extraction of the plants with various buffers, clarification of the suspensions with chloroform or Triton X-100, high speed centrifugation of the virus through sucrose cushion and resuspension of the sedimented virus is different buffers. The two optimal combinations of different procedures tested were either (1) extraction of the plants with the buffer containing 0.3% ascorbic, 0.3% mercaptoethanol, 0.01 mol/l diethyl pyrocarbonate (DEPC) and 5 mmol/l phenylmethylsulphonyl fluoride (PMSF), pH 5.3, clarification with cold chloroform, PEG precipitation, high speed centrifugation through sucrose cushion and resuspension of the sedimented virus in 0.02 mol/l Na-borate buffer pH 7.8, or (2) extraction of the plants with the buffer containing 0.5 mol/l Na-citrate, 1% mercaptoethanol and 5 mmol/l PMSF pH 6.5, clarification with 2% Triton X-100, PEG precipitation, high speed centrifugation through sucrose cushion and resuspension of the sedimented virus in 0.02 mol/l K-phosphate, 0.5 mol/l urea and 0.1% mercaptoethanol, pH 7.5. PMID- 9199717 TI - Some characteristics of a smooth type lipopolysaccharide of Chlamydia psittaci. PMID- 9199716 TI - Phylogenetic analysis of human immunodeficiency virus 1 in Ghana. AB - Eleven human immunodeficiency virus 1 (HIV-1) isolates from Ghanaian acquired immunodeficiency syndrome (AIDS) or AIDS-related complex (ARC) patients obtained by our serosurvey in 1986-1994 were genomically analyzed and phylogenetically compared with other known strains. A phylogenetic tree constructed by analyzing the env region indicated that heterogeneous HIV-1 strains were circulating in Ghana and the majority of them (9 of 11 isolates) belonged to clade (subtype) A which is now furiously epidemic in Africa. Another isolate (1 of 11) belonged to clade D, and the remaining one (1 of 11) belonged to "clade G". This "clade G" virus grouped by the env analysis belonged to clade A by its pol sequence, suggesting an A/G intersubtype recombinant. The characteristic sequences in the V3 tip which have not yet been reported were observed in these Ghanaian isolates, which should be taken into account for future vaccine programs. PMID- 9199718 TI - Failure of Aedes aegypti to become infected by feeding on dengue virus-infected immuno-comprised nude mice. PMID- 9199719 TI - Picobirnavirus, a novel group of undescribed viruses of mammals and birds: a minireview. AB - Picobirnavirus, a novel group of viruses recently detected in children and several species of animals including chickens, are different from the existing members of the family Birnaviridae. Picobirnavirus (PBV) is the tentatively proposed name for the group of these viruses. These viruses are 30-40 nm in diameter and have icosahedral symmetry with triangulation number (T) equal to 3. Their buoyant density in CsCl is 1.4 g/ml. Their genome is bi- or trisegmented double-stranded RNA (dsRNA) with segment lengths of 2.6 and 1.9 kbp for bisegmented and 2.9, 2.4 and 0.9 kbp for trisegmented genomes. The electrophoretic migration profile has considerable heterogeneity. PVBs are detected in diarrhoeic as well as non-diarrhoeic animals, hence, their potential needs further investigation. PMID- 9199720 TI - Parenteral thiamine and Wernicke's encephalopathy: the balance of risks and perception of concern. AB - Wernicke's encephalopathy, a disorder with significant mortality and high morbidity, is common amongst alcohol-dependent patients. Thiamine deficiency appears to play a key role in its aetiology, and parenteral high-dose thiamine is effective in prophylaxis and treatment. Unfortunately, reports of rare anaphylactoid reactions have led to a dramatic reduction in the use of parenteral thiamine, and it is possible that this change in treatment has led, or will lead, to an increase in morbidity and mortality. There is a need for education of doctors who treat alcohol-dependent patients, in order to ensure appropriate use of parenteral thiamine in prophylaxis and treatment of this disorder. PMID- 9199721 TI - Maternal alcohol consumption and spontaneous abortion. AB - This review examines the relationship between maternal alcohol consumption during pregnancy and spontaneous abortions. Although very high spontaneous abortion rates have been reported for alcoholic women, it is still uncertain if this is due to the direct effects of alcohol or the indirect effects of alcoholism related disorders such as cirrhosis. The higher rates of spontaneous abortion among alcoholics may also be due to their higher pregnancy rates. Studies in animals indicate that blood alcohol levels > 200 mg/dl can directly precipitate spontaneous abortion. The association between lower levels of maternal alcohol consumption and spontaneous abortion is much less clear. There is a definite effect of study site in these latter studies: those conducted in North America nearly always report statistically significant associations; those conducted in Europe or Australia nearly always report no significant associations. The reason for this difference is not related to differences in alcohol consumption. Possible explanations for this geographical difference include difference in the socioeconomic status of the women being studied and artefacts associated with the designs used to study these relationships. PMID- 9199722 TI - Ethanol-mediated promotion of oesophageal carcinogenesis: association with lipid peroxidation and changes in phospholipid fatty acid profile of the target tissue. AB - Ethanol consumption is a high risk factor for oesophageal carcinoma and studies indicate that it acts as a promoter of N-nitrosomethylbenzylamine (NMBzA)-induced oesophageal carcinogenesis. The studies described here indicate that ethanol induced promotion was related with an increase in indices of lipid peroxidation in the target oesophageal tissue and that such an increase was associated with significant changes in the fatty acid profile of phospholipids. Young Sprague Dawley rats were treated with NMBzA, 2.5 mg/kg body weight, three times a week for 3 weeks, and a week afterwards fed a 7% ethanolic diet that was continued until their death at 10 months. Cumulative ethane exhaled by rats was measured a week before their death and was found to increase significantly with NMBzA treatment but more so when followed by ethanol consumption. Cholesterol, phospholipids, and some indices of lipid peroxidation were measured in the oesophagus and liver. Whereas the levels of cholesterol and phospholipids were not affected in control-fed rats with or without the NMBzA treatment, ethanol consumption by either the untreated or NMBzA-treated rats caused a significant increase in the targeted oesophagus as well as the liver, the major site of ethanol and carcinogen metabolism. Ethanol consumption also increased all the indices of lipid peroxidation, i.e. malondialdehyde, lipid fluorescence, diene- and triene-conjugates; the largest increases were observed in rats that received both NMBzA and ethanol. A comparison of the fatty acid profile of phospholipids from the oesophagus and liver indicated significant alterations both with the NMBzA treatment and ethanol consumption. However, the fatty acid profile with regard to its peroxidability was significantly modified only with ethanol consumption and only in the oesophagus of the NMBzA-treated or untreated rats. Also, hepatic phospholipids showed a substantial increase in linolenate and no change in arachidonate, but the oesophageal phospholipids exhibited a pronounced increase in the levels of C18:3, C20:2, C20:3, C20:3' and C22:6 with a significant increase in arachidonate when use of ethanol followed the NMBzA treatment, suggesting a disorder in lipid and eicosanoid metabolism. We propose that ethanol may promote carcinogenesis through excessive cell proliferation induced by disordered lipid and eicosanoid metabolism that may cause a selective outgrowth of the initiated cells. PMID- 9199723 TI - The effects of family history, sobriety length, and drinking history in younger alcoholics on P300 auditory-evoked potentials. AB - Event-related potentials (ERPs) have been shown to be different between alcoholics and non-alcoholics. Of particular interest to investigators has been the P300 wave. Because it has been shown that alcohol-induced neural damage can alter P300 waves, particularly amplitude, we attempted to examine alcoholics who most likely suffered little damage because they drank heavily for relatively few years (mean = 6.9 years). The effects of long-term sobriety (mean = 5.0) were also investigated to determine if cognitive functioning, as measured by auditory evoked P300 waves, varies with increased abstinence. Because family history for alcoholism has also been shown to influence P300 amplitude and latency, alcoholics and controls with and without family history were examined. The alcoholic group had significantly longer latencies in P300 measures in both the family history positive and negative groups; P300 amplitudes between alcoholics and non-alcoholics did not vary, regardless of family history. P300 waves were unaffected by sobriety length or drinking history. The results support the hypothesis that P300 differences can be seen between alcoholics and those at risk for alcoholism. PMID- 9199724 TI - The relationship between helping alliance and outcome in outpatient treatment of alcoholics: a comparative study of psychiatric treatment and multimodal behavioural therapy. AB - During the last decades, a positive relation between a good alliance and a successful therapy outcome has been demonstrated across a variety of different therapeutic modalities. The relationship between alliance and drinking outcomes in long-term treatment of alcoholics (12 months or more) has not, as far as we know, been presented. In the present study, alcoholics were randomized to two outpatient treatment programmes; multimodal behavioural therapy (MBT) and psychiatric treatment based on a psychodynamic approach (PT). As part of the study, analyses were performed concerning differences in alliance between the two treatment models (MBT, n = 17; PT, n = 18), and concerning the relationship between alliance and treatment outcome. A Swedish version of the Helping Alliance Questionnaire was used to measure alliance. All therapy sessions were tape recorded. An independent researcher rated the patient's and therapist's contribution to the alliance at the third session (early alliance). Early patient and therapist alliances were not related to sociodemographic data or initial measures of alcohol severity, psychiatric symptoms, or personality structure in either therapy. Early therapist alliance was better in MBT in comparison with PT. For MBT patients, a significant positive correlation was found between early patient alliance and mood dimensions at 6 months. There were no significant positive correlations between early alliance and drinking outcome during the course of treatment and in the third year after start of treatment. Mood and drinking outcome also showed low correlations. In conclusion, an initial positive alliance seems insufficient to reduce alcohol misuse. The relationship between early alliance and improvement in alcohol misuse needs to be further investigated. PMID- 9199725 TI - Israeli Arab and Jewish youth knowledge and opinion about alcohol warning labels: pre-intervention data. AB - This article presents baseline data on the opinion toward alcohol beverage warning labels and on levels of knowledge of the risks discussed in the contents of the labels prior to the labels' introduction, and on levels of knowledge of additional alcohol-related hazards not included in the proposed warning labels, among a sample of 3065 adolescents of four religions living in the northern region of Israel. About 2220 Arab participants (Moslems, Christians and Druze) and 845 Jewish respondents answered in the winter of 1996 a Hebrew version of an American questionnaire, which had been used to measure levels of knowledge of the label in the United States. More respondents were in favour of warning labels on alcohol containers than on advertisements. Arabs as a group were more in favour of warning labels on alcohol containers than Jews. The initial knowledge levels among the participants were not very high, especially concerning the item 'Drinking impairs the ability to operate machinery' (74.4%) which is included on the proposed warning label, and concerning two hazards which are not included: 'Drinking increases risk of cancer' (54.6%) and 'Drinking increases risk of high blood pressure' (60.4%). Abstainers knew more than drinkers that 'Pregnant women should not drink', 'Drinking increases risk of cancer' and 'Alcohol in combination with other drugs is hazardous'. Implications for public health are discussed and alternative warning messages that might be used to inform the Israeli public of several less well-known hazards are suggested. PMID- 9199726 TI - Serum gamma-glutamyl-transpeptidase isoforms in alcoholic liver disease. AB - gamma-Glutamyltranspeptidase (gamma GT) appears in serum in multiple forms; their significance and clinical utility in hepatobiliary and pancreatic diseases are still a matter of controversy. Electrophoretic separation of the multiple forms of gamma GT on agarose gel was performed in 20 alcoholic patients (six with cirrhosis and 14 with fatty liver) and the results compared with those obtained in 50 healthy volunteers, 43 patients affected with chronic hepatitis C, 36 patients with posthepatitic cirrhosis and in 52 epileptic patients on long-term anti-epileptic medication. Multiple forms of gamma GT were separated into several bands (up to 11), labelled 0a, 0b, 1a, 1b, 2a, 2b, 2c, 3a, 3b, 4a, 4b. In the alcoholic patients nine fractions were detected, and the electrophoretic pattern observed was significantly different from that observed in healthy volunteers and in patients with chronic hepatitis C or posthepatitic cirrhosis. No differences were observed in the electrophoretic patterns in the alcohol abusers and epileptic patients. In alcoholic patients significant differences were observed in the electrophoretic patterns in relation to the degree of liver injury; the electrophoretic patterns in patients with alcohol-related cirrhosis and posthepatitic cirrhosis differed significantly. The separation of multiple forms of gamma GT has high sensitivity and good reproducibility. It may be proposed as a complementary test in the diagnosis of alcoholic liver disease. PMID- 9199727 TI - Factors in childhood and youth predicting alcohol dependence and abuse in Swedish women: findings from a general population study. AB - The aim was to assess risk factors during childhood and youth for alcohol dependence/abuse (ADA) in a population-based study of Swedish women. A total of 316 women were interviewed after stratified random sampling in the general population and a screening questionnaire. The interviews focused on social, psychological and behaviour characteristics as well as on early substance use patterns. Alcohol diagnoses were made according to DSM-III-R and CIDI-SAM. Experiences of sexual abuse before the age of 13 years, a history of psychological or psychiatric problems, early deviant behaviour and an episode of alcohol intoxication before the age of 15 years were significantly associated with ADA in a logistic model. General indicators of low social class were not associated with increased risk of ADA in a multivariate analysis. Sexual abuse in childhood was the strongest predictor of ADA. This association has potential public health importance, and should be addressed in future studies on women and alcohol. PMID- 9199728 TI - The effects of alcohol on eye movements during reading. AB - In an experimental double-blind placebo study of 18 subjects (mean age 26.2 years), we investigated the effect of three blood alcohol concentrations (0.0%, 0.05% and 0.1%) on five visuomotor reading parameters: (1) number of eye fixations per 100 words read; (2) the number of words read per minute; (3) the number of regressions per 100 words read; (4) the saccadic length; (5) the duration of eye fixations. The number of fixations and the duration of eye fixations increased significantly as a function of increased breath alcohol concentration. There were no significant changes in the other visuomotor reading parameters. PMID- 9199729 TI - Increasing incidence of Korsakoff's psychosis in the east end of Glasgow. AB - A retrospective analysis of all admissions between 1990 and 1995 in a population of 160,000 identified 47 new cases of Korsakoff's psychosis only seven of which were preceded by Wernicke's encephalopathy. There was a higher ratio of females to males, relative to admissions for severe alcohol dependence. It postulated that the increasing incidence may be related to the warning of anaphylaxis and subsequent withdrawal of high-potency parenteral multivitamins with thiamine. PMID- 9199730 TI - Does liver dysfunction explain neuropsychological status in recently detoxified alcohol-dependent clients? AB - In the search for explanation of persistent cognitive impairment associated with alcohol dependence, the possible role of liver disease has aroused considerable interest. However, review of the relevant literature provides only ambiguous support for any general relationship between neuropsychological status and laboratory tests of liver function. We tested the general hypothesis, and also two specific hypotheses relating particular liver function parameters (gamma glutamyl transferase and serum albumin) to mental ability in a sample of 54 recently detoxified alcohol-dependent people. Despite adequate design power, we failed to obtain evidence for general or specific correlations between mental ability and liver function. We conclude that the accumulated data do not provide direct support for the hypothesis that liver disease plays a part in the genesis of chronic alcohol-related brain impairment in clients without cirrhosis. PMID- 9199731 TI - No longer disabled: the legal impact of the new social construction of HIV. PMID- 9199732 TI - Genetic predispositions, prophylactic treatments and private health insurance: nothing is better than a good pair of genes. PMID- 9199733 TI - Constitutional aspects of physician-assisted suicide after Lee v. Oregon. PMID- 9199734 TI - Narrowing provider choice: any willing provider laws after New York Blue Cross v. Travelers. PMID- 9199735 TI - Developing issues under the Massachusetts 'Physician Profile' Act. PMID- 9199736 TI - Art or science? PMID- 9199737 TI - The revolution in molecular genetics and its impact on ophthalmology. AB - Rapid advances in molecular technology have led to the identification of many genes responsible for inherited disease in ophthalmology. These discoveries also portend an understanding of the pathogenesis of more common ophthalmic disorders which have a genetic component, such as open-angle glaucoma and age-related macular degeneration. This review comprises a summary of these advances in molecular genetics, particularly the contribution of the Human Genome Project; a tabulation of the genes recently proven to be mutated in hereditary ocular conditions; and a discussion of the implications for the practising ophthalmologist. PMID- 9199738 TI - Norman MacAlister Gregg Lecture. The pathogenesis of diabetic retinopathy. AB - Diabetic retinopathy remains a major cause of loss of vision. The Diabetes Control and Complications Trial (DCCT) has implicated hyperglycaemia as a probable major direct causative factor in the pathogenesis of diabetic retinopathy. There are several plausible mechanisms by which high glucose concentrations could lead to the functional and later structural changes characterising diabetic retinopathy. These include increased activity of the aldose reductase pathway, increase de novo synthesis of diacylglycerol from glucose, causing protein kinase C activation, increased non-enzymatic glycation and increased oxidative damage. The demonstration of the potential roles of these pathways and the subsequent effects of growth factors in enhancing angiogenesis provide potential new approaches to the prevention and treatment of diabetic retinopathy. PMID- 9199739 TI - Ida Mann Lecture. Transduction in human photoreceptors. AB - Phototransduction (the process by which light triggers a neural response in retinal rod and cone photoreceptors) is now understood at a molecular level. Indeed, the G-protein cascade of phototransduction is one of the best understood of all biological signalling pathways. The diffusional interactions of the proteins underlying the cascade are described and are briefly analysed. In response to a single activated rhodopsin (R*), formed as a result of a single photon hit, it can be shown that molecules of the G-protein will be activated (to G*) at an approximately constant rate. This, in turn, will cause the number of activated molecules of the third protein (the effector protein, E*, a phosphodiesterase) also to rise linearly with time. These kinetics of protein activation lead to an accurate description of the time-course of the rising phase of the photoreceptor's electrical response, both in single-cell recordings and also in recordings of the human electroretinogram (ERG). By analysing the a-wave of the ERG it is possible to determine the 'amplification' of transduction within living photoreceptors, and to begin to localise the molecular site of dysfunction is cases of photoreceptor abnormality. PMID- 9199740 TI - A clinical trial of MK-507, Trusopt, for raised intraocular pressure--the Australian experience. AB - BACKGROUND: Since the introduction of carbonic anhydrase inhibitors, a topical preparation has been sought to avoid the complications of systemic medication while retaining the effect of lowering intraocular pressure. Recently, a topical carbonic anhydrase inhibitor, MK-507, has been found superior to others and introduced for multicentre clinical trial. PATIENTS AND METHODS: As part of an international multicentre trial, we compared MK-507 with beta blockers for one year in 20 patients with raised intraocular pressure, both as monotherapy and in combination. RESULTS: Twelve patients with a mean peak pressure of 31.9 +/- 6.8 mmHg (range, 22 to 49 mmHg) off all medication received MK-507. After two weeks, mean peak pressure was 24.7 +/- 6.1 mmHg (range, 14 to 41 mmHg)--a 22.6% fall in pressure. Six of these patients had a mean peak pressure of 27.8 +/- 6.4 mmHg (range, 21 to 41 mmHg), a fall of 19.2% from day one and were given timolol as add-on therapy. This resulted in a fall to 19.1 +/- 2.8 (range, 15 to 24 mmHg) at year one, a fall of 31.3% after add-on. Four patients on timolol and four on betaxolol gave similar falls in pressure with an additional fall when MK-507 was used as add-on therapy. CONCLUSIONS: MK-507 demonstrated its effectiveness as an ocular hypotensive agent in this trial of patients with an unusually high rise in pressure. It showed a hypotensive effect roughly equivalent to beta blockers. It is likely that either a topical carbonic anhydrase inhibitor or a cardioselective beta blocker will be the medication of first choice in newly diagnosed glaucoma. PMID- 9199741 TI - The effectiveness of sub-Tenon's infiltration of local anaesthesia for cataract surgery. AB - PURPOSE: To determine if adequate anesthesia and akinesia could be obtained using an inferonasal quadrant sub-Tenons anaesthesia for cataract surgery. METHODS: The sub-Tenons method of local anaesthesia was used in 50 patients undergoing extracapsular cataract extraction and lens implantation. The technique following was that described by JD Stevens in his study of 50 patients. Posterior sub Tenons space was approached through a conjunctival incision in the inferonasal quadrant and the anaesthetic solution delivered by an irrigating cannula. The patients were assessed for residual ocular movements just before surgery. Effectiveness of anaesthesia was assessed during surgery using a verbal pain rating score. Scoring was based on the concept of a visual analogue pain score chart. RESULTS: Total akinesia was obtained in 20% patients and total anaesthesia in 24% patients. The remainder of the patients had adequate akinesia and anaesthesia to proceed with and complete the surgery. CONCLUSION: This method provides satisfactory anaesthesia for cataract surgery. PMID- 9199742 TI - Orbital colour Doppler imaging in carotid-cavernous sinus fistula. AB - BACKGROUND: Colour Doppler imaging (CDI) is a recent advance in ultrasonography that allows for colour-encoded blood flow data of a vascular structure to be displayed simultaneously on a conventional real-time gray-scale B mode image. Real time A and B mode ultrasonography have been used for diagnostic evaluation of ophthalmic disorders since the early 1960s. The haemodynamic characteristics of the ophthalmic circulation have recently been studied by the use of CDI. METHOD: We present three cases of carotid-cavernous sinus fistulas with different presentations. In each case, orbital CDI was used in evaluating the patient's condition. RESULTS: Orbital CDI was successful in confirming the diagnosis in all three cases. CDI was capable of showing the haemodynamic changes in the orbital vasculature which resulted from carotid-cavernous sinus fistula. CONCLUSION: This non-invasive technique presents as an excellent alternative to invasive vascular studies such as angiography for the diagnosis and evaluation of carotid-cavernous sinus fistulas. PMID- 9199743 TI - Excimer phototherapeutic keratectomy for recurrent granular dystrophy. AB - PURPOSE: Corneal grafting is a standard treatment for visually disabling granular dystrophy. The visual results of this procedure are generally good, but can be marred by recurrences of granular deposits some years later. We report the results of phototherapeutic keratectomy (PTK) for recurrent granular dystrophy in three eyes from two patients and discuss the possibilities of treating de novo granular dystrophy with excimer laser. METHODS: The records of two patients (three eyes) treated with excimer PTK for recurrent granular deposits on the donor cornea were reviewed. RESULTS: In all cases visual acuity was improved and repeat corneal grafting avoided. No significant complications occurred although one eye had further recurrence of granular deposits which was also successfully retreated with excimer PTK. Follow-up on these cases varies between five months and three years. CONCLUSIONS: We believe that excimer PTK offers a simple, safe and repeatable way of restoring visual acuity to most cases of recurrent granular dystrophy. Visual recovery is fast and all the incumbent problems of repeat grafting are avoided. PMID- 9199744 TI - Evaluation of topical 0.03% flurbiprofen drops in the treatment of herpetic stromal keratitis. AB - PURPOSE: The management protocol for herpetic stromal keratitis (HSK) is still controversial. We have attempted to compare the relative efficacy of topical dexamethasone 0.01% and flurbiprofen 0.03% in combination with topical acyclovir 3% in HSK. METHODS: In this institutional, prospective, randomized, controlled, double-blind study, 45 clinically diagnosed cases of HSK were randomly distributed into three coded treatment groups--topical placebo, dexamethasone 0.01%, and flurbiprofen 0.03% each in tapering frequency and in combination with acyclovir 3% ointment five times per day for four weeks. Therapeutic response was assessed every third day for four weeks. Decoding of the treatment groups was done at the conclusion of the study and data analysed. RESULTS: Four-week success rate was 93.3% (14 of 15) in the dexamethasone-acyclovir treatment group, 66.7% (10 of 15) in the flurbiprofen-acyclovir treatment group and 20% (3 of 15) in the placebo-acyclovir treatment group. CONCLUSION: While dexamethasone in combination with acyclovir gives the best results in HSK with minimal side-effects, the role of topical flurbiprofen seems promising. PMID- 9199745 TI - Disturbance of central vision after carbon monoxide poisoning. AB - BACKGROUND: Cerebral achromatopsia is a disturbance of colour perception which may be complete or partial. CLINICAL RECORD: A 28-year-old male patient presented five months after carbon monoxide poisoning with achromatopsia. The achromatopsia was unaccompanied by an inability to recognise faces (prosopagnosia) nor was there any disorder of form or depth perception. RESULTS: Magnetic resonance imaging showed bilateral sharply defied areas of haemorrhagic infarction in the globus pallidus with extensive infarction involving temporal and occipital lobes and with apparent partial sparing of the visual cortex, presumably due to arterial insufficiency. The disturbance of central colour vision resolved spontaneously after a further period of 6 months. CONCLUSION: The symptom of achromatopsia is analysed with particular reference to the recent work of Professor Zeki on disturbance of central colour vision following CO poisoning and the unusual MRI findings. PMID- 9199746 TI - Macula toxicity after intravitreal amikacin. AB - BACKGROUND: Although intravitreal aminoglycosides have substantially improved visual prognosis in endophthalmitis, macular infarction may impair full visual recovery. METHODS: We present a case of presumed amikacin retinal toxicity following treatment with amikacin and vancomycin for alpha-haemolytic streptococcal endophthalmitis. RESULTS: Endophthalmitis resolved with improvement in visual acuity to 6/24 at three months. Fundus fluorescein angiography confirmed macular capillary closure and telangiectasis. CONCLUSIONS: Currently accepted intravitreal antibiotic regimens may cause retinal toxicity and macular ischaemia. Treatment strategies aimed at avoiding retinal toxicity are discussed. PMID- 9199747 TI - Vogt-Koyanagi-Harada syndrome in patients of Vietnamese ancestry. AB - BACKGROUND: Vogt-Koyanagi-Harada syndrome (VKH), is an idiopathic, multisystem inflammatory disorder primarily involving the eye. HLA typing has shown a strong association between the HLA-DR4 antigen and people of Japanese, Han Chinese and Hispanic ancestry with VKH. This study reviewed the clinical features and HLA typing of Vietnamese patients with VKH. PATIENTS AND METHODS: A retrospective review of four unrelated Vietnamese patients with VKH seen in private practice and hospital clinic. The American Uveitis Society (1978) criteria for VKH diagnosis were satisfied. Standard microcytotoxic assays for Class I antigens and HLA-DNA typing of Class II DR antigens (DRB1 genotyping) by the PCR-SSO method were performed. RESULTS: The clinical features of VKH in Vietnamese were comparable to those seen in other Asian races. HLA-DR4 was present in three of the four VKH patients. Two of these patients also expressed the allele DRB1*0405. DISCUSSION: The strong association between HLA-DR4 and the DRB1*0405 allele and VKH seen in Japanese people, may well also exist in Vietnamese people. The HLA association suggests an immunogenetic predisposition to VKH. PMID- 9199748 TI - Behcet's disease: corneal perforation as an ocular manifestation. AB - PURPOSE: Although ocular manifestations are common among patients with Behcet's disease, corneal perforation has not been reported in the literature. We report an unusual case of Behcet's disease with corneal perforation as the main ocular involvement. METHODS/RESULTS: An elderly Chinese patient was referred for eye examination because of clinical suspicion of Behcet's disease. Eye examination showed evidence of long-standing autoimmune disease of the eye, mild iritis and a corneal perforation in the left eye. Despite evidence of chronicity and corneal perforation, the patient was asymptomatic. The perforation was successfully treated with cyanoacrylate glue. CONCLUSIONS: While anterior segment involvement is common in Behcet's disease, this case highlights an unusual and hitherto unreported ocular involvement. PMID- 9199749 TI - Oedipism. AB - BACKGROUND: Oedipism or self-enucleation is a rare form of self-mutilation, and most often described in acutely psychotic patients, who have religious or sexual delusions. CLINICAL RECORD: A 47-year-old Chinese man with a history of chronic schizophrenia enucleated his own left eye and mutilated his right eye. The reason for his behaviour was unknown. The history and legends surrounding autoenucleation and the medical literature are reviewed. RESULTS: The patient was managed jointly with the psychiatrist. He suffered extensive injury to his right eye, resulting in loss of vision. CONCLUSION: The management of Oedipism requires close cooperation between ophthalmologists and psychiatrists. Precautions must be taken to prevent repeated attempts or other self-mutilatory behaviour. PMID- 9199750 TI - Long-term complications of Strampelli's osteo-odonto-keratoprosthesis. PMID- 9199751 TI - Blindness and trachoma in South Australia. PMID- 9199752 TI - GP screening for glaucoma. PMID- 9199753 TI - Dendritic cells in the treatment of cancer. PMID- 9199754 TI - Allospecific CD4+, Th1/Th2 and CD8+, Tc1/Tc2 populations in murine GVL: type I cells generate GVL and type II cells abrogate GVL. AB - Donor CD4+ and CD8+ T cells mediate graft-vs.-leukemia (GVL) responses in the allogeneic bone marrow transplantation (alloBMT) setting. To evaluate the role of functional T cell subsets in the mediation of GVL, alloreactive donor CD4+ (Th1/Th2) and CD8+ (Tc1/Tc2) T cells of defined cytokine phenotype were generated by in vitro culture. A leukemia/transplantation model (B6 into B6C3F1; 1050 cGy host irradiation) was established using the bcr/abl-transfected myeloid leukemia line, 32Dp210 (P210; H-2k). Leukemia control mice (1X10(4) P210 cells per recipient) died at day 12.0 post-BMT. Recipients of the CD4+, Th1-type or CD8+, Tc1-type populations were conferred a survival advantage (death at 20.7 and 23.5 days post-BMT, respectively). In contrast, the CD4+, Th2-type population did not mediate GVL (death at 12.3 days). Furthermore, cell mixing experiments demonstrated that the Th2 subset abrogated both Th1- and Tc1-mediated GVL. The CD8+, Tc2 population, which secreted type II cytokines and lysed the P210 leukemia target in vitro, mediated GVL in some experiments; interestingly, the magnitude of Tc2-mediated GVL was inversely related to the level of interleukin 10 (IL-10) secreted in vitro by the Tc2 population. These studies therefore indicate that alloreactive T cells of type I phenotype maximally generate GVL, and that type I/type II interactions are an important consideration for allogeneic transplantation in the setting of leukemic hosts. PMID- 9199755 TI - Long-term engraftment, graft-vs.-host disease, and immunologic reconstitution after experimental transplantation of allogeneic peripheral blood cells from G CSF-treated donors. AB - Peripheral blood cells (PBPC) are an alternative source of bone marrow for allogeneic transplantation. Reports from recent clinical trials granulocyte colony-stimulating factor (G-CSF)-mobilized PBPC for allogeneic transplantation show incidence and severity of graft-vs.-host disease (GVHD) similar to those observed in conventional bone marrow transplantation (BMT), despite the presence of 10- to 20-fold more T cell in the PBPC inoculum. In the present study, we examined the effects of pretreatment of donors with G-CSF on GVHD, long-term engraftment, and lymphocyte reconstitution in a murine parent-->F1 model (B6.Ly 5a-->B6d2F1) using splenocytes as a source of peripheral progenitor cells. Recipients of splenocytes from G-CSF-treated donors experienced less mortality from acute GVHD and showed sustained weight gain by day 100 after transplantation. At that time, there was no histological evidence od GVHD in either liver or gut. Recipients of splenocytes from G-CSF-treated donors showed complete donor engraftment within 1 month, which was sustained until the end of the observation period. In contrast, recipients of T cell-depleted splenocytes showed slower donor engraftment and persistent donor/host chimerism. In addition, lymphocyte phenotype and function in mice receiving splenocytes from G-CSF treated donors was significantly restored by day 100 after transplantation. Thus, the use of G-CSF-mobilized PBPC may provide significant advantages to conventional BMT by reducing GVHD without impairing long-term engraftment and immunologic reconstruction. PMID- 9199757 TI - Novel approaches in blood and marrow transplantation. Report on the 2nd annual scientific meeting of The American Society for Blood and Marrow Transplantation. Joint Special Conference with the American Association for Cancer Research. Jointly sponsored by the College of Physicians and Surgeons of Columbia University. October 2-6, 1996, San Diego, CA. PMID- 9199756 TI - Computer program to predict likelihood of finding and HLA-matched donor: methodology, validation, and application. AB - Approximately 65% of the patients requiring bone marrow transplantation do not have an HLA-A, -B, -DR identical sibling and therefore need to find a phenotypically matched unrelated donor. As of June 30, 1996, the National Marrow Donor Program maintains a registry of 2.31 million volunteer donors, 35% of whom are fully typed for HLA-A, -B, -DR loci. Because a majority of the donors has not been DR typed, a patient who does not find a complete match at the time of the preliminary search may elect to prospectively DR type A, B matched and A, B one antigen mismatched donors. An efficient strategy is therefore needed for determining the likelihood that an appropriate donor exists and for deciding which of the donors that have not yet been DR typed should be tested for DR matching with the candidate. We developed a mathematical algorithm and computer program to facilitate the search for a suitable donor by donor race and phenotype. The program provides information on the likelihood of 1) finding at least one HLA-A, -B, -DR phenotypically matched donor, 2) the likelihood of finding at least one DR match among m A, B matched donors who have not yet been DR typed, and 3) the likelihood of an A, B one antigen mismatched donor of a specific phenotype being a DR match with the patient. The mathematical models underlying the program are based on basic population genetics theory and utilize HLA-A, -B, DR haplotype frequencies derived from the NMDP registry. The results of the validation study show that the prediction is highly accurate at the level of broad antigens. The algorithm and program have the potential to assist patients and physicians in optimizing their decisions regarding clinical management and resource allocation on the process of searching for a suitable unrelated bone marrow donor. PMID- 9199758 TI - Controlled protein delivery from biodegradable tyrosine-containing poly(anhydride co-imide) microspheres. AB - Polymer microspheres capable of the controlled release of macromolecules for periods ranging from days to over a month were developed. The microspheres were made using a new family of anhydride polymers: tyrosine-containing poly(anhydride co-imides), specifically poly[trimellitylimido-L-tyrosine-co-sebacic acid-co-1,3 bis(carboxphenoxy)propane] anhydrides [poly(TMA-Tyr:SA:CPP)]. These polymers may be of particular interest for controlled delivery of vaccine antigens due to the incorporation of an immunological adjuvant, L-tyrosine, into their backbone. Microspheres were produced from a variety of polymer compositions using a double emulsion solvent-evaporation technique, and tested for their ability to provide controlled release of a model protein, bovine serum albumin, in vitro. The microspheres are spherical with smooth surfaces and encapsulate greater than 70% of the protein. Protein release rates from polymers of identical composition could be varied from 0.3 to over 125 micrograms per mg spheres per month by changing the amount of protein encapsulated. This effect can be magnified by using polymers with various monomer ratios. A close correlation between protein release and polymer weight loss was observed, suggesting a release mechanism controlled mainly by polymer erosion. Bovine serum albumin release from poly(TMA Tyr:SA:CPP) microspheres is also pH sensitive, being enhanced at high pH and depressed under acidic conditions. PMID- 9199759 TI - alpha-Amylase and salivary albumin adsorption onto titanium, enamel and dentin: an in vivo study. AB - In vivo adsorption of salivary albumin and alpha-amylase onto titanium, enamel and dentin was analysed following their exposure to the oral cavity for 2h. Oral appliances in six adults served as a platform for carrying 4-mm discs of the three materials. Adherent proteins were eluted from the discs and the amounts of salivary albumin and alpha-amylase were measured by an enzyme-linked immunosorbent assay. While significant difference between the adsorption of albumin and alpha-amylase onto enamel as compared with dentin was observed, adsorption onto titanium was significantly lower. A sample of whole saliva was also collected from each participant. The mean total amounts of albumin and alpha amylase in the participants' whole saliva were 0.03 and 0.54 mg ml-1, respectively. Titanium adsorbed significantly less (4.43%) of the total albumin than did enamel (14.30%) or dentin (18.80%). No significant difference was found in the relative amounts of alpha-amylase adsorbed by the three materials. This significantly selective adsorption of proteins may enable the attachment of specific bacteria and thus alter the composition of the dental plaque and its potential pathogenicity. PMID- 9199760 TI - Experimental evaluation of a resorbable intramedullary plug for cemented total hip replacement. AB - In order to evaluate degradation kinetics and biocompatibility of a resorbable poly(D,L-lactic acid) (PDLLA) plug for total cemented hip prostheses, an experimental in vitro and in vivo study was carried out. Degradation rate studies were performed in Ringer solution and after in vivo plug implantation in the femoral medullar cavity of rabbits. In vitro biocompatibility was evaluated in murine fibroblast cell cultures. PDLLA plugs showed faster degradation kinetics in vivo than in vitro. Histological evaluations showed that PDLLA completely disappeared in vivo 26 weeks after implantation. Fibrous tissue in the medullar cavity was observed at 13 weeks, but no histological changes were observed after 26 weeks. Also, the in vitro tests showed good biocompatibility of the biomaterial. Our results show the possibility of considering this resorbable plug for clinical situations instead of the traditionally used plugs [polyethylene, poly(methyl methacrylate) or cancellous bone] due to its biocompatibility, degradation properties and simplicity of use. PMID- 9199761 TI - Degradation behaviour of a new bioceramic: Ca2P2O7 with addition of Na4P2O7.10H2O. AB - A newly produced bioceramic, beta-Ca2P2O7 with addition of Na4P2O7.10H2O (SDCP), has been implanted into the femoral condyle of rabbits. Within 6 weeks after implantation, most of the bioceramic is replaced by new woven bone. On the contrary, block from hydroxyapatite (HA) and beta-tricalcium phosphate (beta TCP), which are osteoconductible, do not resorb within a short period of time. We believe that the biodegradable behaviour of SDCP may occur in two steps. The first and most important step is the digestion of particles and migration of the particles by phagocytosis. The object of this study is to examine the change in morphologies, chemical compositions and crystal structure of SDCP after soaking in distilled water for a certain period of time. The SDCP ceramic was also co cultured with leucocytes to observe how the SDCP particles were digested by the leucocytes, so that the mechanism of biodegradable behaviour of SDCP ceramic in vivo might be clarified. Four types of sintered calcium phosphate ceramics were tested in the experiment: SDCP, pure beta-Ca2P2O7 (DCP), HA and beta-TCP. They wee soaked in distilled water at 37 degrees C for up to 30 days. The microstructure and morphology of crystals deposited on the surface were observed using scanning electron microscopy. Sodium, calcium and phosphorus ion contents in the supernatant solution were detected by atomic absorption analysis and ion coupled plasma. In summary, HA and DCP showed no significant evidence of dissolution in distilled water. In static distilled water, calcium ions may be released from beta-TCP into solution during the initial 7 days and then converted into HA by reprecipitation. The results showed that the SDCP was firstly dissolved into small grains or fragments by the solution. The small fragments should be so small as to be digested by the phagocytes in a physiological environment. PMID- 9199762 TI - Hydroxyapatite ceramics with selected sintering additives. AB - Several sintering additives for hydroxyapatite (HA) have been tested in order to enhance its sinterability without decomposing the HA and/or decreasing bioactivity and biocompatibility, additionally providing a weak interface for HA ceramics. The ion species of sintering additives were selected from those in the mineral constituents of hard tissues and bioactive glasses. After investigation of phase diagrams in the CaO-P2O5-additive systems, and analysis of physiochemical properties of the additives, several sintering aids for HA have been chosen. Subsequently, densification, phase composition, grain growth and fracture behaviour of HA containing 5 wt% of each additive, sintered at 1000-1100 degrees C, have been studied. H3BO3, CaCl2, KCl, KH2PO4, (KPO3)n and Na2Si2O5 did not enhance densification of HA. K2CO2, Na2CO3, KF and sodium phosphates improved the densification significantly. Expect for KCl and some sodium phosphates, all the additives caused formation of large quantities of undesired beta-tricalcium phosphate or CaO; therefore, they are not appropriate for HA. In the case of sodium phosphate additives, it was possible to avoid formation of CaO or beta tricalcium phosphate by control of the additive quantity and chemical composition. beta-NaCaPO4 has been found to be an effective sintering agent which causes neither decomposition of HA nor formation of other undesired phases. PMID- 9199764 TI - Effect of addition of palladium on properties of Ag2Hg3 (gamma 1) phase. AB - The effect of palladium addition on the microstructure, compressive strength, creep rate and mercury release rate of Ag2Hg3 (gamma 1) phase was evaluated. Experimental results indicated that fairly pure gamma 1 phase could be fabricated using the present trituration method. The heat treatment of gamma 1 at 90 degrees C increased porosity level, increased dimensional shrinkage, increased mercury vapour release and enhanced the formation of beta1 phase. Addition of palladium in gamma 1 slowed down the amalgamation reaction, largely suppressed the phase transition to beta1 and caused a slight shift in open circuit potential toward the anodic direction. Although the overall anodic polarization profiles did not show a significant effect of palladium, scanning electron microscopy revealed morphological differences between pure and palladium-containing gamma 1. Addition of palladium in gamma 1 also increased compressive strength, increased creep resistance, and largely reduced both mercury vapour and ion release rates. Considering overall performance, the optimal palladium content in gamma 1 seems to be in the range between 0.50 and 0.75 wt%. PMID- 9199763 TI - Adhesive composite resins for artificial teeth: a laboratory investigation of bond strength to a cobalt-chromium alloy. AB - Adhesive resin systems are reported to improve the bond strength between resins and cast cobalt-chromium alloy. This investigation compares the behaviour of three resin systems. Cylinders and beams of cobalt-chromium, with 0.6-mm-diameter retention beads regularly cast onto the bonding surfaces, were air-abraded and ultrasonically cleaned. Resin veneers 4 mm deep on the cylinders and 2 mm on the bars were polymerized by heat and pressure or by light. Specimens were water stored for 7 or 90 days, including thermocycling between 4, 37 and 60 degrees C, before testing in a Universal Testing Machine to examine the shear bond strength or the effect of the bonded resin spine on the flexural strength of the beams using a three-point bend test. Specimens were examined with an optical microscope to attempt to determine the nature of the failures that occurred. The investigation showed that, overall, the heat- and pressure-cured urethane dimethacrylate resin with and adhesive based upon methacrylic acid performed significantly less well than a conventional acrylic resin, or a hybrid composite resin with and adhesive monomer. PMID- 9199765 TI - Effects of chitin and its derivatives on the proliferation and cytokine production of fibroblasts in vitro. AB - The effects of chitin and its derivatives on the proliferation of fibroblasts and on the production of cytokines were examined in vitro. Chitin and its derivatives showed almost no acceleratory effect on the proliferation of cultured fibroblasts. On the contrary, high-concentration 500 micrograms ml-1) D glucosamine cultures supplemented with or without a 10% fetal calf serum (FCS) supplementation showed a significant (P < 0.05) reduction in the rate of proliferation of L929 fibroblast cells relative to control. High-concentration chitosan cultures supplemented with 10% FCS showed a significant (P < 0.05) reduction in the rate of L929 fibroblast proliferation. However, the inhibition of cell proliferation by high concentrations of chitosan did not show in cultures without FCS. Interleukin-8 (IL-8) was induced in the supernatants of rat primary cultured dermal fibroblasts stimulated with chitin and its derivatives. Chitin and its derivatives did not stimulate the production of IL-6 by mouse dermal primary cultured fibroblasts. IL-1 alpha, IL-1 beta and tumour necrosis factor alpha were not detected in the fibroblast supernatants. These observations support the notion that cell proliferation is accelerated indirectly by chitin and its derivatives when these materials are used in vivo. In vivo findings of a angiogenesis and migration of neutrophils may be due to persistent release of IL 8 from fibroblasts. PMID- 9199767 TI - Analytical Debye-Huckel model for electrostatic potentials around dissolved DNA. AB - We present an analytical, Green-function-based model for the electric potential of DNA in solution, treating the surrounding solvent with the Debye-Huckel approximation. The partial charge of each atom is accounted for by modeling DNA as linear distributions of atoms on concentric cylindrical surfaces. The condensed ions of the solvent are treated with the Debye-Huckel approximation. The resultant leading term of the potential is that of a continuous shielded line charge, and the higher order terms account for the helical structure. Within several angstroms of the surface there is sufficient information in the electric potential to distinguish features and symmetries of DNA. Plots of the potential and equipotential surfaces, dominated by the phosphate charges, reflect the structural differences between the A, B, and Z conformations and, to a smaller extent, the difference between base sequences. As the distances from the helices increase, the magnitudes of the potentials decrease. However, the bases and sugars account for a larger fraction of the double helix potential with increasing distance. We have found that when the solvent is treated with the Debye-Huckel approximation, the potential decays more rapidly in every direction from the surface than it did in the concentric dielectric cylinder approximation. PMID- 9199768 TI - Collective properties of hydration: long range and specificity of hydrophobic interactions. AB - We report results of molecular dynamics (MD) simulations of composite model solutes in explicit molecular water solvent, eliciting novel aspects of the recently demonstrated, strong many-body character of hydration. Our solutes consist of identical apolar (hydrophobic) elements in fixed configurations. Results show that the many-body character of PMF is sufficiently strong to cause 1) a remarkable extension of the range of hydrophobic interactions between pairs of solute elements, up to distances large enough to rule out pairwise interactions of any type, and 2) a SIF that drives one of the hydrophobic solute elements toward the solvent rather than away from it. These findings complement recent data concerning SIFs on a protein at single-residue resolution and on model systems. They illustrate new important consequences of the collective character of hydration and of PMF and reveal new aspects of hydrophobic interactions and, in general, of SIFs. Their relevance to protein recognition, conformation, function, and folding and to the observed slight yet significant nonadditivity of functional effects of distant point mutations in proteins is discussed. These results point out the functional role of the configurational and dynamical states (and related statistical weights) corresponding to the complex configurational energy landscape of the two interacting systems: biomolecule + water. PMID- 9199766 TI - Transmembrane helix structure, dynamics, and interactions: multi-nanosecond molecular dynamics simulations. AB - To probe the fundamentals of membrane/protein interactions, all-atom multi nanosecond molecular dynamics simulations were conducted on a single transmembrane poly(32)alanine helix in a fully solvated dimyristoyphosphatidylcholine (DMPC) bilayer. The central 12 residues, which interact only with the lipid hydrocarbon chains, maintained a very stable helical structure. Helical regions extended beyond these central 12 residues, but interactions with the lipid fatty-acyl ester linkages, the lipid headgroups, and water molecules made the helix less stable in this region. The C and N termini, exposed largely to water, existed as random coils. As a whole, the helix tilted substantially, from perpendicular to the bilayer plane (0 degree) to a 30 degrees tilt. The helix experienced a bend at its middle, and the two halves of the helix at times assumed substantially different tilts. Frequent hydrogen bonding, of up to 0.7 ns in duration, occurred between peptide and lipid molecules. This resulted in correlated translational diffusion between the helix and a few lipid molecules. Because of the large variation in lipid conformation, the lipid environment of the peptide was not well defined in terms of "annular" lipids and on average consisted of 18 lipid molecules. When compared with a "neat" bilayer without peptide, no significant difference was seen in the bilayer thickness, lipid conformations or diffusion, or headgroup orientation. However, the lipid hydrocarbon chain order parameters showed a significant decrease in order, especially in those methylene groups closest to the headgroup. PMID- 9199769 TI - The renewal of the epidermis: a topological mechanism. AB - Using a topological approach, we study the dynamics of the basement membrane of the mammalian epidermis when basal cells detach or divide. A theoretical characterization of the steady state of the tissue, in very good agreement with experimental data, includes for the first time the division and the disappearance of cells in a two-dimensional random cellular structure. We predict a strong correlation between the size of the attachment of basal cells to the basement membrane and their biological behavior (division or detachment). This suggests that the main factor determining the fate of basal cells, and thus controlling the renewal of the epidermis, is the cells' surface tension and adhesion. PMID- 9199770 TI - Electrophysiological characterization of ionic transport by the retinal exchanger expressed in human embryonic kidney cells. AB - The retinal Na+:Ca2+, K+exchanger cDNA was transiently expressed in human embryonic kidney (HEK 293) cells by transfection with plasmid DNA. The correct targeting of the expressed protein to the plasma membrane was confirmed by immunocytochemistry. The reverse exchange offrent (Ca2+ imported per Na+ extruded) was measured in whole-cell voltage-clamp experiments after intracellular perfusion with Na+ (Na+i, 128 mM) and extracellular perfusion with Ca2+ (Ca2o+, 1 mM) and Ko+ (20 mM). As expected, the exchange current was suppressed by removing Ca2o+. Surprisingly, however, it was also abolished by increasing Na+o to almost abolish the Na+ gradient, and it was almost unaffected by the removal of Ko+. Apparently, then, at variance with the exchanger in the rod outer segment, the retinal exchanger expressed in 293 cells acts essentially as a Na+:Ca2+ exchanger and does not require K+ for its electrogenic activity. PMID- 9199772 TI - Alzheimer's disease amyloid beta-protein forms Zn(2+)-sensitive, cation-selective channels across excised membrane patches from hypothalamic neurons. AB - We have previously shown that the 40-residue peptide termed amyloid beta-protein (A beta P[1-40]) in solution forms cation-selective channels across artificial phospholipid bilayer membranes. To determine whether A beta P[1-40] also forms channels across natural membranes, we used electrically silent excised membrane patches from a cell line derived from hypothalamic gonadotrophin-releasing hormone GnRH neurons. We found that exposing either the internal or the external side of excised membrane patches to A beta P[1-40] leads to the spontaneous formation of cation-selective channels. With Cs+ as the main cation in both the external as well as the internal saline, the amplitude of the A beta P[1-40] channel currents was found to follow the Cs+ gradient and to exhibit spontaneous conductance changes over a wide range (50-500 pS). We also found that free zinc (Zn2+), reported to bind to amyloid beta-protein in solution, can block the flow of Cs+ through the A beta P[1-40] channel. Because the Zn2+ chelator o phenanthroline can reverse this blockade, we conclude that the underlying mechanism involves a direct interaction between the transition element Zn2+ and sites in the A beta P[1-40] channel pore. These properties of the A beta P[1-40] channel are rather similar to those observed in the artificial bilayer system. We also show here, by immunocytochemical confocal microscopy, that amyloid beta protein molecules form deposits closely associated with the plasma membrane of a substantial fraction of the GnRH neurons. Taken together, these results suggest that the interactions between amyloid beta-protein and neuronal membranes also occur in vivo, lending further support to the idea that A beta P[1-40] channel formation might be a mechanism of amyloid beta-protein neurotoxicity. PMID- 9199771 TI - A model of the nicotinic receptor extracellular domain based on sequence identity and residue location. AB - We have modeled the extracellular domains of individual subunits (amino acids 31 200) in the nicotinic acetylcholine receptor using sequence homology with copper binding proteins of known crystal structure, plastocyanin and pseudoazurin, and data from recent site-specific mutagenesis, antibody mapping, and site-directed labelling studies. These data formed an initial model that was refined using molecular dynamics and mechanics as well as electrostatic and solvation energy calculations. The sequences between residues 31 and 164 in the alpha 1-subunit and corresponding residues in homologous receptor subunits show similarity with the core sequence of the cation binding site in plastocyanin and pseudoazurin, a region in the template proteins characterized by multiple hairpin loops. In addition to defining the subunit interfaces that comprise the site for agonist and competitive antagonist binding in more detail, the findings show that negatively charged residues cluster in domains arranged to diminish electrostatic free energy of the complex. Electrostatic factors also appear to distinguish the ligand binding interfaces, alpha gamma and alpha delta, from the other three interfaces on the pentameric receptor. PMID- 9199773 TI - Base-base and deoxyribose-base stacking interactions in B-DNA and Z-DNA: a quantum-chemical study. AB - Base-stacking interactions in canonical and crystal B-DNA and in Z-DNA steps are studied using the ab initio quantum-chemical method with inclusion of electron correlation. The stacking energies in canonical B-DNA base-pair steps vary from 9.5 kcal/mol (GG) to -13.2 kcal/mol (GC). The many-body nonadditivity term, although rather small in absolute value, influences the sequence dependence of stacking energy. The base-stacking energies calculated for CGC and a hypothetical TAT sequence in Z-configuration are similar to those in B-DNA. Comparison with older quantum-chemical studies shows that they do not provide even a qualitatively correct description of base stacking. We also evaluate the base (deoxy)ribose stacking geometry that occurs in Z-DNA and in nucleotides linked by 2',5'-phosphodiester bonds. Although the molecular orbital analysis does not rule out the charge-transfer n-pi* interaction of the sugar 04' with the aromatic base, the base-sugar contact is stabilized by dispersion energy similar to that of stacked bases. The stabilization amounts to almost 4 kcal/mol and is thus comparable to that afforded by normal base-base stacking. This enhancement of the total stacking interaction could contribute to the propensity of short d(CG)n sequences to adopt the Z-conformation. PMID- 9199774 TI - Estimation of the diffusion-limited rate of microtubule assembly. AB - Microtubule assembly is a complex process with individual microtubules alternating stochastically between extended periods of assembly and disassembly, a phenomenon known as dynamic instability. Since the discovery of dynamic instability, molecular models of assembly have generally assumed that tubulin incorporation into the microtubule lattice is primarily reaction-limited. Recently this assumption has been challenged and the importance of diffusion in microtubule assembly dynamics asserted on the basis of scaling arguments, with tubulin gradients predicted to extend over length scales exceeding a cell diameter, approximately 50 microns. To assess whether individual microtubules in vivo assemble at diffusion-limited rates and to predict the theoretical upper limit on the assembly rate, a steady-state mean-field model for the concentration of tubulin about a growing microtubule tip was developed. Using published parameter values for microtubule assembly in vivo (growth rate = 7 microns/min, diffusivity = 6 x 10(-12) m2/s, tubulin concentration = 10 microM), the model predicted that the tubulin concentration at the microtubule tip was approximately 89% of the concentration far from the tip, indicating that microtubule self assembly is not diffusion-limited. Furthermore, the gradients extended less than approximately 50 nm (the equivalent of about two microtubule diameters) from the microtubule tip, a distance much less than a cell diameter. In addition, a general relation was developed to predict the diffusion-limited assembly rate from the diffusivity and bulk tubulin concentration. Using this relation, it was estimated that the maximum theoretical assembly rate is approximately 65 microns/min, above which tubulin can no longer diffuse rapidly enough to support faster growth. PMID- 9199775 TI - Numerical simulation of local calcium movements during L-type calcium channel gating in the cardiac diad. AB - Computer simulation was used to investigate the calcium levels after sarcolemmal calcium influx through L-type calcium channels (DHPRs) into the narrow diadic space of cardiac muscle. The effect of various cytosolic and membranebound buffers, diad geometry, DHPR properties (open time and current), and surface charge were examined. The simulations showed that phospholipid binding sites on the sarcolemmal membrane are the major buffer affecting free calcium ([Ca2+]) levels in the diad. The inclusion of surface charge effects calculated from Gouy Chapman theory resulted in a marked decrease in [Ca2+] levels at all times and a faster decay of [Ca2+] after termination of DHPR influx. For a DHPR current of 200 fA, [Ca2+] at the center of the diad reached peak levels of approximately 73 microM. In larger diads (> or = 400 nm diameter), [Ca2+] decayed more slowly than in smaller diads (100-200 nm diameter), although peak [Ca2+] levels reached during typical DHPR open times were similar. For a wide range of DHPR single channel current magnitudes (Ica = 25-200 fA), [Ca2+] levels in the diad were approximately proportional to ICa. The decrease in calculated [Ca2+] levels due to the effects of surface charge can be interpreted as resulting from an effective "volume expansion" of the diad space. Furthermore, the layer of increased [Ca2+] close to the sarcolemmal membrane can act as a fast buffer. PMID- 9199776 TI - Numerical analysis of ryanodine receptor activation by L-type channel activity in the cardiac muscle diad. AB - Computer simulations were used to examine the response of ryanodine receptors (RyRs) to the sarcolemmal calcium influx via L-type calcium channels (DHPRs). The effects of ryanodine receptor organization, diad geometry, DHPR single-channel current, and DHPR gating were examined. In agreement with experimental findings, the simulations showed that RyRs can respond rapidly (approximately 0.4 ms) to calcium influx via DHPRs. The responsiveness of the RyR depends on the geometrical arrangement between the RyRs and the DHPR in the diad, with wider diads being generally less responsive. When the DHPR single-channel current is small (approximately 25 fA), the organization of RyRs into small clusters results in an improved responsiveness. With experimentally observed DHPR mean open and closed times (0.17 ms and 4 ms, respectively) it is the first opening of the DHPR that is most likely to activate the RyR. A measure of the efficiency (Q) by which DHPR gating evokes sarcoplasmic reticulum release is defined. Q is at maximum for tau approximately 0.3 ms, and we interpret this finding in terms of the "tuning" of DHPR gating to RyR response. If certain cardiac myopathies are associated with a mismatch in the "tuning," then modification of DHPR gating with drugs to "retune" calcium-induced calcium release should be possible. PMID- 9199777 TI - Extension of torsionally stressed DNA by external force. AB - Metropolis Monte Carlo simulation was used to study the elasticity of torsionally stressed double-helical DNA. Equilibrium distributions of DNA conformations for different values of linking deficit, external force, and ionic conditions were simulated using the discrete wormlike chain model. Ionic conditions were specified in terms of DNA effective diameter, i.e., hard-core radius of the model chain. The simulations show that entropic elasticity of the double helix depends on how much it is twisted. For low amounts of twisting (less than about one turn per twist persistence length) the force versus extension is nearly the same as in the completely torsionally relaxed case. For more twisting than this, the molecule starts to supercoil, and there is an increase in the force needed to realize a given extension. For sufficiently large amounts of twist, the entire chain is plectonemically supercoiled at low extensions; a finite force must be applied to obtain any extension at all in this regime. The simulation results agree well with the results of recent micromanipulation experiments. PMID- 9199778 TI - Modeling the electrophoresis of lysozyme. II. Inclusion of ion relaxation. AB - In this work, boundary element methods are used to model the electrophoretic mobility of lysozyme over the pH range 2-6. The model treats the protein as a rigid body of arbitrary shape and charge distribution derived from the crystal structure. Extending earlier studies, the present work treats the equilibrium electrostatic potential at the level of the full Poisson-Boltzmann (PB) equation and accounts for ion relaxation. This is achieved by solving simultaneously the Poisson, ion transport, and Navier-Stokes equations by an iterative boundary element procedure. Treating the equilibrium electrostatics at the level of the full rather than the linear PB equation, but leaving relaxation out, does improve agreement between experimental and simulated mobilities, including ion relaxation improves it even more. The effects of nonlinear electrostatics and ion relaxation are greatest at low pH, where the net charge on lysozyme is greatest. In the absence of relaxation, a linear dependence of mobility and average polyion surface potential, (lambda zero)s, is observed, and the mobility is well described by the equation [formula: see text] where epsilon 0 is the dielectric constant of the solvent, and eta is the solvent viscosity. This breaks down, however, when ion relaxation is included and the mobility is less than predicted by the above equation. Whether or not ion relaxation is included, the mobility is found to be fairly insensitive to the charge distribution within the lysozyme model or the internal dielectric constant. PMID- 9199780 TI - Calcium channel block by (-)devapamil is affected by the sequence environment and composition of the phenylalkylamine receptor site. AB - The pore-forming alpha 1 subunit of L-type calcium (Ca2+) channels is the molecular target of Ca2+ channel blockers such as phenylalkylamines (PAAs). Association and dissociation rates of (-)devapamil were compared for a highly PAA sensitive L-type Ca2+ channel chimera (Lh) and various class A Ca2+ channel mutants. These mutants carry the high-affinity determinants of the PAA receptor site in a class A sequence environment. Apparent drug association and dissociation rate constants were significantly affected by the sequence environment (class A or L-type) of the PAA receptor site. Single point mutations affecting the high-affinity determinants in segments IVS6 of the PAA receptor site, introduced into a class A environment, reduced the apparent drug association rates. Mutation I1811M in transmembrane segment IVS6 (mutant AL25/-I) had the highest impact and decreased the apparent association rate for ( )devapamil by approximately 30-fold, suggesting that this pore-lining isoleucine in transmembrane segment IVS6 plays a key role in the formation of the PAA receptor site. In contrast, apparent drug dissociation rates of Ca2+ channels in the resting state were almost unaffected by point mutations of the PAA receptor site. PMID- 9199779 TI - Heterogeneity of Ca2+ gating of skeletal muscle and cardiac ryanodine receptors. AB - The single-channel activity of rabbit skeletal muscle ryanodine receptor (skeletal RyR) and dog cardiac RyR was studied as a function of cytosolic [Ca2+]. The studies reveal that for both skeletal and cardiac RyRs, heterogeneous populations of channels exist, rather than a uniform behavior. Skeletal muscle RyRs displayed two extremes of behavior: 1) low-activity RyRs (LA skeletal RyRs, approximately 35% of the channels) had very low open probability (Po < 0.1) at all [Ca2+] and remained closed in the presence of Mg2+ (2 mM) and ATP (1 mM); 2) high-activity RyRs (HA skeletal RyRs) had much higher activity and displayed further heterogeneity in their Po values at low [Ca2+] (< 50 nM), and in their patterns of activation by [Ca2+]. Hill coefficients for activation (nHa) varied from 0.8 to 5.2. Cardiac RyRs, in comparison, behaved more homogeneously. Most cardiac RyRs were closed at 100 nM [Ca2+] and activated in a cooperative manner (nHa ranged from 1.6 to 5.0), reaching a high Po (> 0.6) in the presence and absence of Mg2+ and ATP. Heart RyRs were much less sensitive (10x) to inhibition by [Ca2+] than skeletal RyRs. The differential heterogeneity of heart versus skeletal muscle RyRs may reflect the modulation required for calcium-induced calcium release versus depolarization-induced Ca2+ release. PMID- 9199781 TI - Single-channel properties of a rat brain endoplasmic reticulum anion channel. AB - Many intracellular membranes contain ion channels, although their physiological roles are often poorly understood. In this study we incorporated single anion channels colocalized with rat brain endoplasmic reticulum (ER) ryanodine sensitive Ca(2+)-release channels into planar lipid bilayers. The channels opened in bursts, with more activity at negative (cytoplasm-ER lumen) membrane potentials, and they occupied four open conductance levels with frequencies well described by the binomial equation. The probability of a protomer being open decreased from approximately 0.7 at -40 mV to approximately 0.2 at +40 mV, and the channels selected between different anions in the order PSCN > PNO3 > PBr > PCl > PF. They were also permeant to cations, including the large cation Tris+ (PTris/PCl = 0.16). Their conductance saturated at 170 pS in choline Cl. The channels were inactivated by 15 microM 4,4'-diisothiocyanatostilbene-2,2' disulfonic acid (DIDS) and blocked with low affinity (KD of 1-100 microM) by anthracene-9-carboxylic acid, ethacrynic acid, frusemide (furosemide), HEPES, the indanyloxyacetic acid derivative IAA-94, 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB), and Zn2+. Unlike protein translocation pores, the channels were unaffected by high salt concentrations or puromycin. They may regulate ER Ca2+ release, or be channel components en route to their final cellular destinations. Alternatively, they may contribute to the fusion machinery involved in intracellular membrane trafficking. PMID- 9199782 TI - Tail currents in the myelinated axon of Xenopus laevis suggest a two-open-state Na channel. AB - Na tail currents in the myelinated axon of Xenopus laevis were measured at -70 mV after steps to -10 mV. The tail currents were biexponential, comprising a fast and a slow component. The time constant of the slow tail component, analyzed in the time window 0.35-0.50 ms, was independent of step duration, and had a value of 0.23 ms. The amplitude, extrapolated back to time 0, varied, however, with step duration. It reached a peak after 0.7 ms and inactivated relatively slowly (at 2.1 ms the absolute value was reduced by approximately 30%). The amplitude of the fast component, estimated by subtracting the amplitude of the slow component from the calculated total tail current amplitude, reached a peak (three times larger than that of the slow component) after 0.5 ms and inactivated relatively fast (at 2.1 ms it was reduced by approximately 65%). The results were explained by a novel Na channel model, comprising two open states bifurcating from a common closed state and with separate inactivating pathways. A voltage-regulated use of the two pathways explains a number of findings reported in the literature. PMID- 9199783 TI - Comparison of Na+/K(+)-ATPase pump currents activated by ATP concentration or voltage jumps. AB - Using the giant patch technique, we combined two fast relaxation methods on excised patches from guinea pig cardiomyocytes to compare the rate constants of the involved reaction steps. Experiments were done in the absence of intra- or extracellular K+. Fast ATP concentration jumps were generated by photolysis of caged ATP at pH 6.3 with laser flash irradiation at a wavelength of 308 nm and 10 ns duration, as described previously. Transient outward currents with a fast rising phase, followed by a slower decay and a small stationary current, were obtained. Voltage pulses were applied to the same patch in the presence or absence of intracellular ATP. Subtraction of the voltage jump-induced currents in the absence of ATP from those taken in the presence of ATP yielded monoexponential transient current signals, which were dependent on external Na+ but did not differ between intracellular pH (pHi) values 6.3 or 7.4. Rate constants showed a characteristic voltage dependence, i.e., saturating at positive potentials (approximately 200 s-1, 24 degrees C) and exponentially rising with increasing negative potentials. Rate constants of the fast component from transient currents obtained after an ATP concentration jump agree well with rate constants from currents obtained after a voltage jump to zero or positive potentials (pHi 6.3), and the two exhibit the same activation energy of approximately 80 kJ.mol-1. For a given membrane patch, the amount of charge that is moved across the plasma membrane is roughly the same for each of the two relaxation techniques. PMID- 9199784 TI - Topology of the P segments in the sodium channel pore revealed by cysteine mutagenesis. AB - The P segments of the voltage-dependent Na+ channel line the outer mouth and selectivity filter of the pore. The residues that form the cytoplasmic mouth of the pore of the channel have not been identified. To study the structure of the inner pore mouth, the presumed selectivity filter residues (D400, E755, K1237, and A1529), and three amino acids just amino-terminal to each of these residues in the rat skeletal muscle Na+ channel, were mutated to cysteine and expressed in tsA 201 cells. These amino acids are predicted (by analogy to K+ channels) to be on the cytoplasmic side of the putative selectivity filter residues. Inward and outward Na+ currents were measured with the whole-cell configuration of the patch clamp technique. Cysteinyl side-chain accessibility was gauged by sensitivity to Cd2+ block and by reactivity with methanethiosulfonate (MTS) reagents applied to both the inside and the outside of the cell. Outward currents through the wild type and all of the mutant channels were unaffected by internal Cd2+ (100 microM). Similarly, 1 mM methanethiosulfonate ethylammonium (MTSEA) applied to the inside of the membrane did not affect wild-type or mutant outward currents. However, two mutants amino-terminal to the selectivity position in domain III (F1236C and T1235C) and one in domain IV (S1528C) were blocked with high affinity by external Cd2+. The Na+ current through F1236C and S1528C channels was inhibited by MTSEA applied to the outside of the cell. The accessibility of these mutants to externally applied cysteinyl ligands indicates that the side chains of the mutated residues face outward rather than inward. The K+ channel model of the P segments as protein loops that span the selectivity region is not applicable to the Na+ channel. PMID- 9199786 TI - Detection of spontaneous synaptic events with an optimally scaled template. AB - Spontaneous synaptic events can be difficult to detect when their amplitudes are close to the background noise level. Here we report a sensitive new technique for automatic detection of small asynchronous events. A waveform with the time course of a typical synaptic event (a template) is slid along the current or voltage trace and optimally scaled to fit the data at each position. A detection criterion is calculated based on the optimum scaling factor and the quality of the fit. An event is detected when this criterion crosses a threshold level. The algorithm automatically compensates for changes in recording noise. The sensitivity and selectivity of the method were tested using real and simulated data, and the influence of the template parameter settings was investigated. Its performance was comparable to that obtained by visual event detection, and it was more sensitive than previously described threshold detection techniques. Under typical recording conditions, all fast synaptic events with amplitudes of at least three times the noise standard deviation (3 sigma) could be detected, as could 75% of events with amplitudes of 2 sigma. The scaled template technique is implemented within a commercial data analysis application and can be applied to many standard electrophysiological data file formats. PMID- 9199785 TI - The effects of synaptic noise on measurements of evoked excitatory postsynaptic response amplitudes. AB - Spontaneously occurring synaptic events (synaptic noise) recorded intracellularly are usually assumed to be independent of evoked postsynaptic responses and to contaminate measures of postsynaptic response amplitude in a roughly Gaussian manner. Here we derive analytically the expected noise distribution for excitatory synaptic noise and investigate its effects on amplitude histograms. We propose that some fraction of this excitatory noise is initiated at the same release sites that contribute to the evoked synaptic event and develop an analytical model of the interaction between this fraction of the noise and the evoked postsynaptic response amplitude. Recording intracellularly with sharp microelectrodes in the in vitro hippocampal slice preparation, we find that excitatory synaptic noise accounts for up to 70% of the intracellular recording noise, when inhibition is blocked pharmacologically. Up to 20% of this noise shows a significant correlation with the evoked event amplitude, and the behavior of this component of the noise is consistent with a model which assumes that each release site experiences a refractory period of approximately 60 ms after release. In contrast with classical models of quantal variance, our models predict that excitatory synaptic noise can cause the apparent variance of successive peaks in an excitatory synaptic amplitude histogram to decrease from left to right, and in some cases to be less than the variance of the measured noise. PMID- 9199787 TI - Zero-order interfacial enzymatic degradation of phospholipid tubules. AB - The first study of enzymatic hydrolysis of phospholipid tubules is reported. Phosphatidylcholines with acyl chains containing diacetylene groups are known to form tubular microstructures in which the lipids are tightly packed and crystalline. These tubules can be used to probe the role of microstructural form in the mechanics of interfacial enzymatic degradation by such enzymes as phospholipase A2 (PLA2). Hydrolysis by PLA2 may occur most rapidly in regions having the greatest number of bilayer packing defects, such as those that must be found at tubule ends. A microstructure that degrades primarily from its ends should exhibit zero-order kinetics, because the area of the degrading tubule and remains constant as the length of the microstructure decreases. Free fatty acid concentration was measured to follow the generation of PLA2 hydrolysis products in suspensions of diacetylenic phospholipid tubules. The kinetics of tubule hydrolysis were essentially zero-order until conversion was complete, as predicted. However, microscopy of partially hydrolyzed tubules revealed the formation of multiple discrete anionic product domains along the length of degrading tubules as well as in insoluble reaction product microstructures. Furthermore, the rate of tubule hydrolysis was only moderately enhanced by increasing the number of tubule ends, which is consistent with the conclusion that tubule ends are not the only sites of hydrolysis. A model that reconciles the overall kinetics with the morphological evidence is proposed. PMID- 9199789 TI - Adhesion-induced domain formation by interplay of long-range repulsion and short range attraction force: a model membrane study. AB - We study the role of the interplay of specific and universal forces for the adhesion of giant vesicles on solid supported membranes. To model the situation of cell adhesion, we incorporated lipopolymers (phospholipids with polyethyleneoxide headgroups) as artificial glycocalix, whereas attractive lock and-key forces are mimicked by incorporating biotinylated lipids into both membranes and by mediating the strong coupling through streptavidin. Adhesion is studied by quantitative reflection interference contrast microscopy (RICM), which enables visualization of the contact zone and reconstruction of the height profile of the membrane beyond the contact line (outside the contact zone) up to a height of 1 micron. We demonstrate that adhesion is accompanied by lateral phase separation, leading to the formation of domains of tight adhesion (adhesion plaques) separated by areas of weak adhesion exhibiting pronounced flickering. By analyzing the height profile S(x) near the contact line in terms of the tension equilibrium (Young equation) and the moment equilibrium, respectively, the adhesion energy and membrane tension can be approximately measured locally. We show that the adhesion energy is about three orders of magnitude larger for the adhesion plaques than for the weekly adhering regions. The adhesion is studied as a function of the excess area of the vesicle generated by temperature variation. A very remarkable finding is that increased excess area is not always stored in the contact area, but leads to the formation of microbuds (diameter approximately 2 microns). PMID- 9199788 TI - Effect of changing the size of lipid headgroup on peptide insertion into membranes. AB - Adsorption of amphiphilic peptides to the headgroup region of a lipid bilayer is a common mode of protein-membrane interactions. Previous studies have shown that adsorption causes membrane thinning. The degree of the thinning depends on the degree of the lateral expansion caused by the peptide adsorption. If this simple molecular mechanism is correct, the degree of lateral expansion and consequently the membrane thinning should depend on the size of the headgroup relative to the cross section of the hydrocarbon chains. Previously we have established the connection between the alamethicin insertion transition and the membrane thinning effect. In this paper we use oriented circular dichroism to study the effect of varying the size of the headgroup, while maintaining a constant cross section of the lipid chains, on the insertion transition. A simple quantitative prediction agrees very well with the experiment. PMID- 9199790 TI - Effect of polyethyleneglycol-phospholipids on aggregate structure in preparations of small unilamellar liposomes. AB - Phospholipids with covalently attached poly(ethylene glycol) (PEG lipids) are commonly used for the preparation of long circulating liposomes. Although it is well known that lipid/PEG-lipid mixed micelles may form above a certain critical concentration of PEG-lipid, little is known about the effects of PEG-lipids on liposome structure and leakage at submicellar concentrations. In this study we have used cryogenic transmission electron microscopy to investigate the effect of PEG(2000)-PE on aggregate structure in preparations of liposomes with different membrane compositions. The results reveal a number of important aggregate structures not documented before. The micrographs show that enclosure of PEG-PE induces the formation of open bilayer discs at concentrations well below those where mixed micelles begin to form. The maximum concentration of PEG-lipid that may be incorporated without alteration of the liposome structure depends on the phospholipid chain length, whereas phospholipid saturation or the presence of cholesterol has little or no effect. The presence of cholesterol does, however, affect the shape of the mixed micelles formed at high concentrations of PEG lipid. Threadlike micelles form in the absence of cholesterol but adapt a globular shape when cholesterol is present. PMID- 9199791 TI - The influence of cholesterol on phospholipid membrane curvature and bending elasticity. AB - The behavior of dioleoylphosphatidylethanolamine (DOPE)/cholesterol/tetradecane and dioleoylphosphatidylcholine (DOPC)/cholesterol/tetradecane were examined using x-ray diffraction and the osmotic stress method. DOPE/tetradecane, with or without cholesterol, forms inverted hexagonal (HII) phases in excess water. DOPC/tetradecane forms lamellar phases without cholesterol at lower temperatures. With tetradecane, as little as 5 mol% cholesterol in DOPC induced the formation of HII phases of very large dimension. Increasing levels of cholesterol result in a systematic decrease in the HII lattice dimension for both DOPE and DOPC in excess water. Using osmotic pressure to control hydration, we applied a recent prescription to estimate the intrinsic curvature and bending modulus of the HII monolayers. The radii of the intrinsic curvature, RPO, at a pivotal plane of constant area within the monolayer were determined to be 29.4 A for DOPE/tetradecane at 22 degrees C, decreasing to 27 A at 30 mol% cholesterol. For DOPC/tetradecane at 32 degrees C, RPO decreased from 62.5 A to 40 A as its cholesterol content increased from 30 to 50 mol%. These data yielded an estimate of the intrinsic radius of curvature for pure DOPC of 87.3 A. The bending moduli kc of DOPE/tetradecane and DOPC/tetradecane, each with 30 mol% cholesterol, are 15 and 9 kT, respectively. Tetradecane itself was shown to have little effect on the bending modulus in the cases of DOPE and cholesterol/DOPE. Surprisingly, cholesterol effected only a modest increase in the kc of these monolayers, which is much smaller than estimated from its effect on the area compressibility modulus in bilayers. We discuss possible reasons for this difference. PMID- 9199792 TI - Effects of lipid headgroup and packing stress on poly(ethylene glycol)-induced phospholipid vesicle aggregation and fusion. AB - The effect of lipid headgroup and curvature-related acyl packing stress on PEG induced phospholipid vesicle aggregation and fusion were studied by measuring vesicle and aggregate sizes using the quasi-elastic light scattering and fluorescence energy transfer techniques. The effect of the lipid headgroup was monitored by varying the relative phosphatidylcholine (PC) and phosphatidylethanolamine (PE) contents in the vesicles, and the influence of hydrocarbon chain packing stress was controlled either by the relative amount of PE and PC content in the vesicles, or by the degree of unsaturation of the acyl chains of a series of PEs, e.g., dilinoleoylphosphatidylethanolamine (dilin-PE), lysophosphatidylethanolamine (lyso-PE), and transacylated egg phosphatidylethanolamine (TPE). The PEG threshold for aggregation depends only weakly on the headgroup composition of vesicles. However, in addition to the lipid headgroup, the curvature stress of the monolayer that forms the vesicle walls plays a very important role in fusion. Highly stressed vesicles, i.e., vesicles containing PE with highly unsaturated chains, need less PEG to induce fusion. This finding applies to the fusion of both small unilamellar vesicles and large unilamellar vesicles. The effect of electrostatic charge on vesicle aggregation and fusion were studied by changing the pH of the vesicle suspension media. At pH 9, when PE headgroups are weakly charged, increasing electrostatic repulsion between headgroups on the same bilayer surface reduces curvature stress, whereas increasing electrostatic repulsion between apposing bilayer headgroups hinders intervesicle approach, both of which inhibit aggregation and fusion, as expected. PMID- 9199793 TI - Influence of cis double bonds in the sn-2 acyl chain of phosphatidylethanolamine on the gel-to-liquid crystalline phase transition. AB - We have semisynthesized 19 species of mixed-chain phosphatidylethanolamines (PEs) in which the sn-1 acyl chain is derived from saturated fatty acids with varying chain lengths and the sn-2 acyl chain has different chain lengths but contains 0, 1, and 2 cis double bond(s). The gel-to-liquid crystalline phase transition temperatures (Tm) of lipid bilayers prepared from these 19 mixed-chain PEs were determined calorimetrically. When the Tm values are compared with those of saturated and monounsaturated counterparts, a common Tm profile is observed in the plot of Tm versus the number of cis double bonds. Specifically, a marked stepwise decrease in Tm is detected as the number of cis double bonds in the sn-2 acyl chain of the mixed-chain PE is successively increased from 0 to 1 and then to 2. The large Tm-lowering effect of the acyl chain unsaturation can be attributed to the increase in Gibbs free energy of the gel-state bilayer as a result of weaker lateral chain-chain interactions. In addition, we have applied molecular mechanics calculations to simulate the molecular structure of dienoic mixed-chain C(X):C(Y:2 delta n,n+3)PE in the gel-state bilayer, thus enabling the three independent structural parameters (N, delta C, and LS) to be calculated in terms of X, Y, and n, which are intrinsic quantities of C(X):C(Y:2 delta n,n+3)PE. When the Tm values and the corresponding N and delta C values of all dienoic mixed-chain PEs under study are first codified and then analyzed statistically by multiple regressions, the dependence of Tm on the structural parameters can be described quantitatively by a simple and general equation. The physical meaning and the usefulness of this simple and general equation are explained. PMID- 9199794 TI - Characteristics of troponin C binding to the myofibrillar thin filament: extraction of troponin C is not random along the length of the thin filament. AB - Troponin C (TnC) is the Ca(2+)-sensing subunit of troponin responsible for initiating the cascade of events resulting in contraction of striated muscle. This protein can be readily extracted from myofibrils with low-ionic-strength EDTA-containing buffers. The properties of TnC extraction have not been characterized at the structural level, nor have the interactions of TnC with the native myofibrillar thin filament been studied. To address these issues, fluorescein-labeled TnC, in conjunction with high-resolution digital fluorescence microscopy, was used to characterize TnC binding to myofibrils and to determine the randomness of TnC extraction. Fluorescein-5-maleimide TnC (F5M TnC) retained biological activity, as evidenced by reconstitution of Ca(2+)-dependent ATPase activity in extracted myofibrils and binding to TnI in a Ca(2+)-sensitive manner. The binding of F5M TnC to highly extracted myofibrils at low Ca2+ was restricted to the overlap region under rigor conditions, and the location of binding was not influenced by F5M TnC concentration. The addition of myosin subfragment 1 to occupy all actin sites resulted in F5M TnC being bound in both the overlap and nonoverlap regions. However, very little F5M TnC was bound to myofibrils under relaxing conditions. These results suggest that strong binding of myosin heads enhances TnC binding. At high Ca2+, the pattern of F5M TnC binding was concentration dependent: binding was restricted to the overlap region at low F5M TnC concentration, whereas the binding propagated into the nonoverlap region at higher levels. Analysis of fluorescence intensity showed the greatest binding of F5M TnC at high Ca2+ with S1, and these conditions were used to characterize partially TnC-extracted myofibrils. Comparison of partially extracted myofibrils showed that low levels of extraction were associated with greater F5M TnC being bound in the nonoverlap region than in the overlap region relative to higher levels of extraction. These results show that TnC extraction is not random along the length of the thin filament, but occurs more readily in the nonoverlap region. This observation, in conjunction with the influence of rigor heads on the pattern of F5M TnC binding, suggests that strong myosin binding to actin stabilizes TnC binding at low Ca2+. PMID- 9199795 TI - Analysis of birefringence decay profiles for nucleic acid helices possessing bends: the tau-ratio approach. AB - For nucleic acid helices in the 100-200-bp range, a central bend or point of flexibility increases the rate of rotational diffusion. In a transient electric birefringence (TEB) experiment, this increase is manifest as a reduction in the terminal (slowest) birefringence decay time. Previous experimental and theoretical work has demonstrated that the ratio of the decay times for a bent/flexible molecule and its fully duplex (linear) counterpart represents a sensitive, quantifiable measure of the apparent bend angle (tau-ratio approach). In the current work, we have examined the influence of helix parameters (e.g., persistence length, helix rise, diameter) on the tau-ratio for a given bend. The tau-ratio is found to be remarkably insensitive to variations and/or uncertainties in the helix parameters, provided that one employs bent and control molecules with the same sequence and length (apart from the bend itself). Although a single tau-ratio determination normally does not enable one to distinguish between fixed and flexible bends, such a distinction can be made from a set of tau-ratios for molecules possessing two variably phased bends. A number of additional uncertainties are examined, including errors in the estimation of the dimensions of nonhelix elements that are responsible for bends; such errors can, in principle, be estimated by performing a series of measurements for molecules of varying length. PMID- 9199796 TI - Influence of static and dynamic bends on the birefringence decay profile of RNA helices: Brownian dynamics simulations. AB - Bends in nucleic acid helices can be quantified in a transient electric birefringence (TEB) experiment from the ratio of the terminal decay times of the bent molecule and its fully duplex counterpart (tau-ratio method). The apparent bend angles can be extracted from the experimental tau-ratios through the application of static (equilibrium-ensemble) hydrodynamic models; however, such models do not properly address the faster component(s) of the birefringence decay profile, which can represent up to 80% of the total birefringence signal for large band angles. To address this latter issue, the relative amplitudes of the components in the birefringence decay profile have been analyzed through a series of Brownian dynamics (BD) simulations. Decay profiles have been simulated for three-, five-, and nine-bead models representing RNA molecules with central bends of 30 degrees, 60 degrees, and 90 degrees, and with various degrees of associated angle dispersion. The BD simulations are in close agreement with experimental results for the fractional amplitudes, suggesting that both amplitudes and terminal tau-ratios can be used as a measure of the magnitudes of bends in the helix axis. Although the current results indicate that it is generally not possible to distinguish between relatively fixed and highly flexible bends from single tau-ratio measurements, because they can lead to similar reductions in terminal decay time and amplitude, measurements of the dependence of the fractional amplitudes on helix length may afford such a distinction. PMID- 9199797 TI - Influence of sequence on the conformation of the B-DNA helix. AB - We have tried to ascertain whether the variability found in the conformational features of the 10 base steps in B-DNA is mainly due to the flanking sequences or to interactions with the environment. From an analysis of the twist parameter of the base-pair steps available from crystals of oligonucleotides and protein/oligonucleotide complexes, we conclude that in most cases the flanking sequences show little influence: the conformation of a DNA region results from the combination of the independent intrinsic features of each base step (average conformation and intrinsic variability), modulated by their interactions with the environment. Only in some cases (YR steps, in particular CG and CA/TG) does it appear that flanking sequences have an influence on the conformation of the central base step. The values obtained allow an approximation to the parameters expected for repetitive DNA sequences. In particular, it is found that poly[d(AG/CT)] should have a strongly alternating conformation, in agreement with recently reported oligonucleotide structures. PMID- 9199798 TI - Nucleic acid vibrational circular dichroism, absorption, and linear dichroism spectra. I. A DeVoe theory approach. AB - Infrared (IR) vibrational circular dichroism (VCD), absorption, and linear dichroism (LD) spectra of four homopolyribonucleotides, poly(rA), poly(rG), poly(rC), and poly(rU), have been calculated, in the 1750-1550 cm-1 spectral region, using the DeVoe polarizability theory. A newly derived algorithm, which approximates the Hilbert transform of imaginaries to reals, was used in the calculations to obtain real parts of oscillator polarizabilities associated with each normal mode. The calculated spectra of the polynucleotides were compared with previously measured solution spectra. The good agreement between calculated and measured polynucleotide spectra indicates, for the first time, that the DeVoe theory is a useful means of calculating the VCD and IR absorption spectra of polynucleotides. For the first time, calculated DeVoe theory VCD and IR absorption spectra of oriented polynucleotides are presented. The calculated VCD spectra for the oriented polynucleotides are used to predict the spectra for such measurements made in the future. The calculated IR spectra for the oriented polynucleotides are useful in interpreting the linear dichroism of the polynucleotides. PMID- 9199799 TI - Supramolecular self-assembly of d(TGG)4, synergistic effects of K+ and Mg2+. AB - Spectral evidence indicates that molar concentrations of K+ can induce aggregate formation in d(TGG)4. The 320-nm turbidity monitoring indicates that more than 1 M KCl is needed for the onset of aggregation to occur at 20 degrees C within the time span of 24 h. The kinetic profile is reminiscent of autocatalytic reactions that consist of a lag period followed by accelerative and levelling phases. Progressive shortening of lag periods and more rapid accelerative phases accompany further increases in [K+]. Interestingly, the presence of Mg2+ greatly facilitates the aggregate formation and results in the prominent appearance of an intense psi-type CD. For example, whereas 1 M K+ fails to induce aggregate formation of d(TGG)4 within 24 h, the addition of 1 mM Mg2+ to a 1 M K+ solution is sufficient to induce the onset of aggregation in approximately 12 h. Furthermore, adjustment of the buffer to 16 mM Mg2+/1 M KCl reduces the lag time to less than 10 min and aggregation is nearly complete in 2 h. The requirement of [K+] for aggregation is reduced to 2 mM in the presence of 16 mM Mg2+, a reduction of nearly three orders of magnitude when compared to solutions without Mg2+. The effects of K+ and Mg2+ ions are synergistic, because the presence of 16 mM Mg2+ alone does not induce aggregate formation in this oligomer. Thermal stabilities of the aggregates are strongly dependent on the concentrations of these two ions. Although aggregates formed in the presence of 2 M KCl alone melt around 55 degrees C, those formed with added 16 mM Mg2+ melt at approximately 90 degrees C, with some aggregates remaining unmelted even at 95 degrees C. The slow kinetics of aggregate formation led to the appearance of gross hystereses in the cooling profiles. The interplay of these two ions appears to be specific, because the replacement of K+ by Na+ or the replacement of Mg2+ by other divalent cations does not lead to the observed self-assembly phenomenon, although Sr2+ can substitute for K+. A possible mechanism for the formation of self-assembled structures is suggested. PMID- 9199800 TI - Restricted motion of photoexcited bacteriorhodopsin in purple membrane containing ethanol. AB - The molecular motion of retinal within the purple membrane was investigated by flash-induced absorption anisotropies with or without ethanol. In the absence of ethanol, the measured anisotropies at several wavelengths exhibited almost the same slow decay. This slow decay was attributed to only the rotation of purple membrane sheet itself in the aqueous suspension. In the presence of ethanol, however, we observed the wavelength-dependent anisotropies. The fluidity of the purple membrane, investigated with a fluorescence anisotropy method, was increased by the addition of ethanol. These facts indicated that the characteristic motion of bacteriorhodopsin is induced in perturbed purple membrane with ethanol. The data analysis was performed, taking account of the overlapping of absorption from ground-state bacteriorhodopsin and photointermediates. The results showed that the rotational motion of photointermediates within the membrane was more restricted than that of nonexcited bacteriorhodopsin. The addition of ethanol facilitated the rotation of nonexcited protein, whereas it did not significantly affect the motion of photointermediates. The restricted motion of photointermediates is probably caused by a conformational change in them, which may hinder the rotation of monomer protein and/or induce the interaction between photointermediate and neighboring proteins. PMID- 9199801 TI - Kinetics of H+ ion binding by the P+QA-state of bacterial photosynthetic reaction centers: rate limitation within the protein. AB - The kinetics of flash-induced H+ ion binding by isolated reaction centers (RCs) of Rhodobacter sphaeroides, strain R-26, were measured, using pH indicators and conductimetry, in the presence of terbutryn to block electron transfer between the primary and secondary quinones (QA and QB), and in the absence of exogenous electron donors to the oxidized primary donor, P+, i.e., the P+QA-state. Under these conditions, proton binding by RCs is to the protein rather than to any of the cofactors. After light activation to form P+QA-, the kinetics of proton binding were monoexponential at all pH values studied. At neutral pH, the apparent bimolecular rate constant was close to the diffusional limit for proton transfer in aqueous solution (approximately 10(11) M-1 s-1), but increased significantly in the alkaline pH range (e.g., 2 x 10(13) M-1 s-1 at pH 10). The average slope of the pH dependence was -0.4 instead of -1.0, as might be expected for a H+ diffusion-controlled process. High activation energy (0.54 eV at pH 8.0) and weak viscosity dependence showed that H+ ion uptake by RCs is not limited by diffusion. The salt dependence of the H+ ion binding rate and the pK values of the protonatable amino acid residues of the reaction center implicated surface charge influences, and Gouy-Chapman theory provided a workable description of the ionic effects as arising from modulation of the pH at the surface of the RC. Incubation in D2O caused small increases in the pKs of the protonatable groups and a small, pH (pD)-dependent slowing of the binding rate. The salt, pH, temperature, viscosity, and D2O dependences of the proton uptake by RCs in the P+QA- state were accounted for by three considerations: 1) parallel pathways of H+ delivery to the RC, contributing to the observed (net) H+ disappearance; 2) rate limitation of the protonation of target groups within the protein by conformational dynamics; and 3) electrostatic influences of charged groups in the protein, via the surface pH. PMID- 9199802 TI - Probing protein structure by solvent perturbation of NMR spectra: the surface accessibility of bovine pancreatic trypsin inhibitor. AB - In the absence of specific interactions, the relative attenuation of protein NMR signals due to added stable free radicals such as TEMPOL should reflect the solvent accessibility of the molecular surface. The quantitative correlation between observed attenuation and surface accessibility was investigated with a model system, i.e., the small protein bovine pancreatic trypsin inhibitor. A detailed discussion is presented on the reliability and limits of the approach, and guidelines are provided for data acquisition, treatment, and interpretation. The NMR-derived accessibilities are compared with those obtained from x-ray diffraction and molecular dynamics data. Although the time-averaged accessibilities from molecular dynamics are ideally suited to fit the NMR data, better agreement was observed between the paramagnetic attenuations of the fingerprint cross-peaks of homonuclear proton spectra and the total NH and H alpha accessibilities calculated from x-ray coordinates, than from time-averaged molecular dynamics simulations. In addition, the solvent perturbation response appears to be a promising approach for detecting the thermal conformational evolution of secondary structure elements in proteins. PMID- 9199803 TI - Interpretation of multiple Q(0,0) bands in the absorption spectrum of Mg mesoporphyrin embedded in horseradish peroxidase. AB - Mg-mesoporphyrin horseradish peroxidase (MgMP-HRP) and MgMP-HRP complexed with naphtohydroxamic acid (NHA) have been studied by fluorescence line narrowing (FLN) and pressure tuning spectral hole burning (SHB) techniques. In each sample, the low temperature absorption spectra show more than one transition in the origin range of the Q band. Comparisons with broad-band fluorescence spectra and FLN studies suggest that the multiple band feature originates from the presence of different configurations of the metal-porphyrin that are subject to Qx-Qy splitting within the protein cavity. This suggestion is supported by pressure tuning SHB studies. In the uncomplexed as well as in the NHA-complexed form of MgMP-HRP, irradiation in the Q band produces photoproduct bands, which has been attributed to a species with smaller Qx-Qy splitting. In an amorphous matrix, on the other hand, only one form of MgMP could be found, and no splitting could be observed. The binding of NHA does not significantly alter the bulk parameters of the protein matrix, but it reduces the structural variety in the configuration of MgMP to a single form with a more distorted structure and thus with an enlarged Qx-Qy splitting. PMID- 9199804 TI - Secondary structures comparison of aquaporin-1 and bacteriorhodopsin: a Fourier transform infrared spectroscopy study of two-dimensional membrane crystals. AB - Aquaporins are integral membrane proteins found in diverse animal and plant tissues that mediate the permeability of plasma membranes to water molecules. Projection maps of two-dimensional crystals of aquaporin-1 (AQP1) reconstituted in lipid membranes suggested the presence of six to eight transmembrane helices in the protein. However, data from other sequence and spectroscopic analyses indicate that this protein may adopt a porin-like beta-barrel fold. In this paper, we use Fourier transform infrared spectroscopy to characterize the secondary structure of highly purified native and proteolyzed AQP1 reconstituted in membrane crystalline arrays and compare it to bacteriorhodopsin. For this analysis the fractional secondary structure contents have been determined by using several different algorithms. In addition, a neural network-based evaluation of the Fourier transform infrared spectra in terms of numbers of secondary structure segments and their interconnections [sij] has been performed. The following conclusions were reached: 1) AQP1 is a highly helical protein (42 48% alpha-helix) with little or no beta-sheet content. 2) The alpha-helices have a transmembrane orientation, but are more tilted (21 degrees or 27 degrees, depending on the considered refractive index) than the bacteriorhodopsin helices. 3) The helices in AQP1 undergo limited hydrogen/deuterium exchange and thus are not readily accessible to solvent. Our data support the AQP1 structural model derived from sequence prediction and epitope insertion experiments: AQP1 is a protein with at least six closely associated alpha-helices that span the lipid membrane. PMID- 9199805 TI - Hydrolysis of GTP associated with the formation of tubulin oligomers is involved in microtubule nucleation. AB - Hydrolysis of GTP is known to accompany microtubule assembly. Here we show that hydrolysis of GTP is also associated with the formation of linear oligomers of tubulin, which are precursors (prenuclei) in microtubule assembly. The hydrolysis of GTP on these linear oligomers inhibits the lateral association of GTP-tubulin that leads to the formation of a bidimensional lattice. Therefore GTP hydrolysis interferes with the nucleation of microtubules. Linear oligomers are also formed in mixtures of GTP-tubulin and GDP-tubulin. The hydrolysis of GTP associated with heterologous interactions between GTP-tubulin and GDP-tubulin in the cooligomer takes place at a threefold faster rate than upon homologous interactions between GTP-tubulins. The implication of these results in a model of vectorial GTP hydrolysis in microtubule assembly is discussed. PMID- 9199806 TI - Compartmentalized energy transfer in cardiomyocytes: use of mathematical modeling for analysis of in vivo regulation of respiration. AB - The mathematical model of the compartmentalized energy transfer system in cardiac myocytes presented includes mitochondrial synthesis of ATP by ATP synthase, phosphocreatine production in the coupled mitochondrial creatine kinase reaction, the myofibrillar and cytoplasmic creatine kinase reactions, ATP utilization by actomyosin ATPase during the contraction cycle, and diffusional exchange of metabolites between different compartments. The model was used to calculate the changes in metabolite profiles during the cardiac cycle, metabolite and energy fluxes in different cellular compartments at high workload (corresponding to the rate of oxygen consumption of 46 mu atoms of O.(g wet mass)-1.min-1) under varying conditions of restricted ADP diffusion across mitochondrial outer membrane and creatine kinase isoenzyme "switchoff." In the complete system, restricted diffusion of ADP across the outer mitochondrial membrane stabilizes phosphocreatine production in cardiac mitochondria and increases the role of the phosphocreatine shuttle in energy transport and respiration regulation. Selective inhibition of myoplasmic or mitochondrial creatine kinase (modeling the experiments with transgenic animals) results in "takeover" of their function by another, active creatine kinase isoenzyme. This mathematical modeling also shows that assumption of the creatine kinase equilibrium in the cell may only be a very rough approximation to the reality at increased workload. The mathematical model developed can be used as a basis for further quantitative analyses of energy fluxes in the cell and their regulation, particularly by adding modules for adenylate kinase, the glycolytic system, and other reactions of energy metabolism of the cell. PMID- 9199807 TI - Interaction of rabbit C-reactive protein with phospholipid monolayers studied by microfluorescence film balance with an externally applied electric field. AB - C-reactive protein (CRP) is one of the most characteristic acute-phase proteins in humans and many other animals. It binds to phosphorylcholine in a calcium dependent manner. In addition, CRP activates the complement systems via the classical pathway. The interaction between rabbit CRP (rCRP) and model biological membrane is studied using dimyristoylphosphatidylethanolamine and dipalmitoylphosphatidylcholine monolayers. Observations with fluorescence microscopy indicate that rCRP is more likely to be incorporated in the liquid phase of monolayers. Such incorporation does not depend on the presence of calcium and is not inhibited by phosphocholine. The area occupied by the protein when incorporated into the monolayer was estimated. The dipole moment density of the protein crossing the air/water interface was measured by applying an external electric field. Our results indicate that calcium binding leads to a conformational change in CPR, which might modify the orientation of CRP in the monolayer. In addition, a negative charge or negative difference in dipole moment density facilitates the incorporation of CPR into the monolayer. PMID- 9199808 TI - Analysis of phosphorescence in heterogeneous systems using distributions of quencher concentration. AB - A continuous distribution approach, instead of the traditional mono- and multiexponential analysis, for determining quencher concentration in a heterogeneous system has been developed. A mathematical model of phosphorescence decay inside a volume with homogeneous concentration of phosphor and heterogeneous concentration of quencher was formulated to obtain pulse-response fitting functions for four different distributions of quencher concentration: rectangular, normal (Gaussian), gamma, and multimodal. The analysis was applied to parameter estimates of a heterogeneous distribution of oxygen tension (PO2) within a volume. Simulated phosphorescence decay data were randomly generated for different distributions and heterogeneity of PO2 inside the excitation/emission volume, consisting of 200 domains, and then fit with equations developed for the four models. Analysis using a monoexponential fit yielded a systematic error (underestimate) in mean PO2 that increased with the degree of heterogeneity. The fitting procedures based on the continuous distribution approach returned more accurate values for parameters of the generated PO2 distribution than did the monoexponential fit. The parameters of the fit (M = mean; sigma = standard deviation) were investigated as a function of signal-to-noise ratio (SNR = maximum signal amplitude/peak-to-peak noise). The best-fit parameter values were stable when SNR > or = 20. All four fitting models returned accurate values of M and sigma for different PO2 distributions. The ability of our procedures to resolve two different heterogeneous compartments was also demonstrated using a bimodal fitting model. An approximate scheme was formulated to allow calculation of the first moments of a spatial distribution of quencher without specifying the distribution. In addition, a procedure for the recovery of a histogram, representing the quencher concentration distribution, was developed and successfully tested. PMID- 9199809 TI - Photoacoustic analysis of proteins: volumetric signals and fluorescence quantum yields. AB - A series of proteins has been examined using time-resolved, pulsed-laser volumetric photoacoustic spectroscopy. Photoacoustic waveforms were collected to measure heat release for calculation of fluorescence quantum yields, and to explore the possibility of photoinduced nonthermal volume changes occurring in these protein samples. The proteins studied were the green fluorescent protein (GFP); intestinal fatty acid binding protein (IFABP), and adipocyte lipid-binding protein (ALBP), each labeled noncovalently with 1-anilinonaphthalene-8-sulfonate (1,8-ANS) and covalently with 6-acryloyl-2-(dimethylamino)naphthalene (acrylodan); and acrylodan-labeled IFABP and ALBP with added oleic acid. Of this group of proteins, only the ALBP labeled with 1,8-ANS showed significant nonthermal volume changes at the beta = 0 temperature (approximately 3.8 degrees C) for the buffer used (10 mM Tris-HCI, pH 7.5) (beta is the thermal cubic volumetric expansion coefficient). For all of the proteins except for acrylodan labeled IFABP, the fluorescence quantum yields calculated assuming simple energy conservation were anomalously high, i.e., the apparent heat signals were lower than those predicted from independent fluorescence measurements. The consistent anomalies suggest that the low photoacoustic signals may be characteristic of fluorophores buried in proteins, and that photoacoustic signals derive in part from the microenvironment of the absorbing chromophore. PMID- 9199810 TI - High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators. AB - The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging. PMID- 9199812 TI - Polarized light scattering for rapid observation of bacterial size changes. AB - The effect of changing growth conditions on the diameter of rod-shaped bacteria was studied in vivo with the use of polarized light scattering. The value of a ratio of scattering matrix elements was measured as a function of scattering angle at various times after nutritional "upshift" for two strains of Escherichia coli cells. The peak locations of the scattering function were calibrated against the diameter for rod-shaped bacteria. The peaks moved toward smaller angles as a function of time after upshift, indicating that the diameter was increasing. Under special conditions, substantial peak shifts occurred within a few minutes of growth condition change, indicating a rapid onset of growth in diameter. The rate of increase of the diameters after upshift was obtained from the angular shift of peak location. This rate was approximately 14 nm/min for E. coli K12 and approximately 9 nm/min for E. coli B/r at 37 degrees C. The rate of diameter increase is smaller at lower temperatures. Experiments with Bacillus megaterium showed that any diameter change after nutritional upshift at 37 degrees C is limited to at most a very small increase, at least for the strain and medium tested. PMID- 9199811 TI - Structure and orientation of lung surfactant SP-C and L-alpha dipalmitoylphosphatidylcholine in aqueous monolayers. AB - SP-C, a pulmonary surfactant-specific protein, aids the spreading of the main surfactant phospholipid L-alpha-dipalmitoylphosphatidylcholine (DPPC) across air/water interfaces, a process that has possible implications for in vivo function. To understand the molecular mechanism of this process, we have used external infrared reflection-absorption spectroscopy (IRRAS) to determine DPPC acyl chain conformation and orientation as well as SP-C secondary structure and helix tilt angle in mixed DPPC/SP-C monolayers in situ at the air/water interface. The SP-C helix tilt angle changed from approximately 24 degrees to the interface normal in lipid bilayers to approximately 70 degrees in the mixed monolayer films, whereas the acyl chain tilt angle of DPPC decreased from approximately 26 degrees in pure lipid monolayers (comparable to bilayers) to approximately 10 degrees in the mixed monolayer films. The protein acts as a "hydrophobic lever" by maximizing its interactions with the lipid acyl chains while simultaneously permitting the lipids to remain conformationally ordered. In addition to providing a reasonable molecular mechanism for protein-aided spreading of ordered lipids, these measurements constitute the first quantitative determination of SP-C orientation in Langmuir films, a paradigm widely used to simulate processes at the air/alveolar interface. PMID- 9199813 TI - Barriers to diffusion of plasma membrane proteins form early during guinea pig spermiogenesis. AB - The plasma membrane of the mature guinea pig sperm is segregated into at least four domains of different composition. Previous studies have shown that some proteins localized within these domains are free to diffuse laterally, suggesting that barriers to protein diffusion are responsible for maintaining the nonuniform distribution of at least some surface proteins in mature sperm. The different membrane domains appear sequentially during sperm morphogenesis in the testis and during later passage through the epididymis. To determine when diffusion barriers become functional during sperm development, we examined the diffusion of two proteins that are expressed on the cell surface of developing spermatids and become segregated to different plasma membrane domains during the course of spermiogenesis. Both proteins exhibited rapid lateral diffusion throughout spermiogenesis, even after they become localized to specific regions of the surface membrane. These results suggest that barriers to membrane diffusion form concomitantly with membrane domains during spermiogenesis. PMID- 9199814 TI - Simulation of detachment of specifically bound particles from surfaces by shear flow. AB - The receptor-mediated adhesion of cells to ligand-coated surfaces is important in many physiological and biotechnological processes. Previously, we measured the detachment of antibody-coated spheres from counter-antibody- and protein A-coated substrates using a radial-flow detachment assay and were able to relate mechanical adhesion strength to chemical binding affinity (Kuo and Lauffenburger, Biophys. J. 65:2191-2200 (1993)). In this paper, we use "adhesive dynamics" to simulate the detachment of antibody-coated hard spheres from a ligand-coated substrate. We modeled the antibody-ligand (either counter-antibody or protein A) bonds as adhesive springs. In the simulation as in the experiments, beads attach to the substrate under static conditions. Flow is then initiated, and detachment is measured by the significant displacement of previously bound particles. The model can simulate the effects of many parameters on cell detachment, including hydrodynamic stresses, receptor number, ligand density, reaction rates between receptor and ligand, and stiffness and reactive compliance of the adhesive springs. The simulations are compared with experimental detachment data, thus relating measured bead adhesion strength to molecular properties of the adhesion molecules. The simulations accurately recreated the logarithmic dependence of adhesion strength on affinity of receptor-ligand recognition, which was seen in experiments and predicted by analytic theory. In addition, we find the value of the reactive compliance, the parameter which relates the strain of a bond to its rate of breakage, that gives the best match between theory and experiment to be 0.01. Finally, we analyzed the effect of varying either the forward or reverse rate constants as different ways to achieve the same affinity, and showed that adhesion strength depends uniquely on the equilibrium affinity, not on the kinetics of binding. Given that attachment is independent of affinity, detachment and attachment are distinct adhesive phenomena. PMID- 9199815 TI - Kinetic studies of Ca2+ binding and Ca2+ clearance in the cytosol of adrenal chromaffin cells. AB - The Ca2+ binding kinetics of fura-2, DM-nitrophen, and the endogenous Ca2+ buffer, which determine the time course of Ca2+ changes after photolysis of DM nitrophen, were studied in bovine chromaffin cells. The in vivo Ca2+ association rate constants of fura-2, DM-nitrophen, and the endogenous Ca2+ buffer were measured to be 5.17 x 10(8) M-1 s-1, 3.5 x 10(7) M-1 s-1, and 1.07 x 10(8) M-1 s 1, respectively. The endogenous Ca2+ buffer appeared to have a low affinity for Ca2+ with a dissociation constant around 100 microM. A fast Ca2+ uptake mechanism was also found to play a dominant role in the clearance of Ca2+ after flashes at high intracellular free Ca2+ concentrations ([Ca2+]), causing a fast [Ca2+]i decay within seconds. This Ca2+ clearance was identified as mitochondrial Ca2+ uptake. Its uptake kinetics were studied by analyzing the Ca2+ decay at high [Ca2+]i after flash photolysis of DM-nitrophen. The capacity of the mitochondrial uptake corresponds to a total cytosolic Ca2+ load of approximately 1 mM. PMID- 9199816 TI - Force generation in the outer hair cell of the cochlea. AB - The outer hair cell of the mammalian cochlea has a unique motility directly dependent on the membrane potential. Examination of the force generated by the cell is an important step in clarifying the detailed mechanism as well as the biological importance of this motility. We performed a series of experiments to measure force in which an elastic probe was attached to the cell near the cuticular plate and the cell was driven with voltage pulses delivered from a patch pipette under whole-cell voltage clamp. The axial stiffness was also determined with the same cell by stretching it with the patch pipette. The isometric force generated by the cell is around 0.1 nN/mV, somewhat smaller than 0.15 nN/mV, predicted by an area motor model based on mechanical isotropy, but larger than in earlier reports in which the membrane potential was not controlled. The axial stiffness obtained, however, was, on average, 510 nN per unit strain, about half of the value expected from the mechanical isotropy of the membrane. We extended the area motor theory incorporating mechanical orthotropy to accommodate the axial stiffness determined. The force expected from the orthotropic model was within experimental uncertainties. PMID- 9199817 TI - Evaluation of a new automated ELISA test for von Willebrand factor using two monoclonal antibodies. AB - A new automated quantitative enzyme linked immunosorbent assay (ELISA) for the detection of human von Willebrand factor (vWF), VIDAS vWF, has been developed for use on the VIDAS analyser (bioMerieux). The two-step capture/tag test relies on two monoclonal antibodies (mAbs), the second one being labelled with alkaline phosphatase. The lower limit of detection of the assay is < 1 IU/dl, and the upper limit of detection is 120 IU/dl. There is no hook effect for concentrations up to 1100 IU/dl. Intra- and inter-assay precision ranges from 3% and 5%. Assays are performed preferentially using citrated plasma and in these conditions the 95% confidence intervals for normal values are 52-154 IU/dl and 60-200 IU/dl for O blood group and non-O blood group subjects, respectively. Using the lower limits of the normal range as the cut-off level, all patients tested with type 1 (n = 29) or 3 (n = 2) von Willebrand disease (vWD) would be accurately classified with the new ELISA. Comparing the VIDAS test with a conventional ELISA gave a good correlation and comparable results with type 1 and 3 vWD patients (n = 31; r = 0.99; y = 0.99x + 0.24), type 2A and 2B vWD patients (n = 20; r = 0.99; y = 1.05x-1.65) and normal subjects (n = 204; r = 0.94; y = 1.06x-2.6). PMID- 9199819 TI - Activation of coagulation and platelets is affected by the hydrophobicity of artificial surfaces. AB - Clot formation is a limiting factor in the use of biomaterials. We investigated the effect of surface hydrophobicity on haemostatic activation in vitro, using five polyetherurethanes of varying surface hydrophobicity (C94, C74, C54 and C34), C94 the most, and C34, the least hydrophobic, and compared them with a commercial standard pellethane. Sterilised sacks were filled with heparinised blood, rotated at 37 degrees C for 24 h and sequential samples collected into 0.103 M sodium citrate. Thrombin generation measured by thrombin-antithrombin III complexes showed a difference between the polymers at 3 h through to 6 h (P < 0.05), C94 showing the least activation and C34 the most. Factor XIIa and D-dimer levels increased between 12 (P < 0.05) and 24 h (P < 0.01) for all polyetherurethanes. The ratio of soft:hard segments (which determine hydrophobicity) of the polyetherurethanes showed a direct relationship with the degree of activation of coagulation and fibrinolysis. There was no significant increase in monocyte tissue factor expression at 5 and 105 min. Platelet function as measured by whole blood platelet aggregation showed a reduction with pellethane and C94 after 1 h using collagen, with no changes for C34, Altering surface hydrophobicity has diverse effects on haemostatic pathways, with the most hydrophobic surfaces causing least activation of coagulation but most activation of platelets. PMID- 9199818 TI - Fibrinogenolysis and thrombin generation after reduced dose bolus or conventional rt-PA for pulmonary embolism. The Coagulation Project Investigators of the Bolus Alteplase Pulmonary Embolism Group. AB - The aim of this study was to compare the effects on fibrinogenolysis and thrombin generation of two recombinant tissue-type plasminogen activator (rt-PA) regimens in patients with pulmonary embolism entering a randomised, controlled study with a 1:2 allocation ratio to rt-PA, 100 mg over 2 h (Group A) or rt-PA, 0.6 mg/kg, maximum dose 50 mg, over 15 min (Group B). In both groups the heparin infusion was stopped 2-4 h before starting thrombolytic treatment and resumed accordingly to the activated partial thromboplastin time (aPTT) or thrombin clotting time (TcT). Seventeen patients in Group A and 30 patients in Group B were evaluated before starting thrombolytic treatment and 2, 6 and 24 h after its end for the following parameters: aPTT, TcT, fibrinogen, fibrinogen degradation products (FDP), plasmin-alpha 2 antiplasmin (PAP) and thrombin-antithrombin III (TAT) complexes. The two groups had similar coagulation parameters at baseline. Two h after starting rt-PA, the aPTT was more prolonged in Group A than in Group B patients (P = 0.01). Patients in Group B showed less reduction in plasma fibrinogen levels at all study times after rt-PA treatment (P = 0.008). The increase in plasma FDP (P = 0.037) and PAP (P = 0.001) levels was lower at 2 and 6 h samples in Group B compared with Group A. TcT was prolonged (P = 0.003) and TAT increased (P = 0.001) during treatment without differences between the two groups. AUC0-24 of fibrinogen, FDP and PAP levels confirmed the statistically significant differences (P = 0.04) between the two groups over the entire 24 h period of the study. Three patients in Group A (17.6%) and three (10.0%) in Group B suffered major or other important bleeding. Our results indicate that the administration of weight-adjusted reduced-dose rt-PA bolus produces less impairment of blood coagulation than the FDA approved regimen. PMID- 9199820 TI - Acquired deficiency of antithrombin in association with a hypercoagulable state and impaired function of liver and/or kidney in preeclampsia. AB - To determine whether decreases in plasma antithrombin (AT) level, as seen in non gestational acquired AT deficiency, result from a hypercoagulable state and/or liver/kidney damage, AT activity was measured in 24 uncomplicated and 30 preeclamptic women. The fifth percentile of AT levels in the normal pregnancies was used as a cut-off value to subdivide the preeclamptic patients into two groups. Markers of activated coagulation, i.e, levels of thrombin-antithrombin complex (TAT), fibrin D-dimer, soluble fibrin, von Willebrand factor (vWF) and platelet counts, were determined. Indicators of hepatic or renal function, i.e. concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine, urinary albumin (U-albumin) and serum albumin (S-albumin), were assayed. AT levels were lower in those with preeclampsia than in the normal pregnancy group (P < 0.01). In the group with AT levels less than the cut-off point, levels of fibrin D-dimer (P < 0.05), soluble fibrin (P < 0.05), vWF (P < 0.05), ALT (P < 0.05), AST (P < 0.05), creatinine (P < 0.01) and U-albumin (P < 0.01) were increased, whereas platelet counts (P < 0.05) and S-albumin (P < 0.05) were decreased. All patients with ALT levels > 0.46 mu kat/1, AST > 0.58 mu kat/1, S-albumin < 23 g/1 and/or U-albumin > 4.9 g/24 h had AT levels < or = cut off. AT levels correlated with vWF (rs = - 0.73, P < 0.01) and creatinine (Rs = 0.70, P < 0.01). It is suggested that in preeclampsia, acquired AT deficiency is secondary to a hypercoagulable state, and/or associated with impaired hepatic and/or renal function. PMID- 9199821 TI - Variability of the response to activated protein C during normal pregnancy. AB - The response to activated protein C (APC) was investigated in 28 healthy women, non-carriers of the Arg506-Gln mutation in factor V, throughout pregnancy (gestation weeks 12, 20, 28, 32 and 37) and after the delivery. A suppression of APC response was observed which reached lowest values by week 28 (nAPC-ratio 0.78 +/- 0.13), sustained low up to the end of pregnancy and rose after delivery (1.11 +/- 0.22; P < 0.05). APC resistance (nAPC ratio < 0.75) was registered in 16 of the 28 women (57%). A reduction of APC ratio was directly related to its value in the non-pregnant state, being most pronounced in the women with the highest APC ratio. Factor VIII increased during pregnancy and correlated inversely to APC ratio (Z coefficient = -0.645, P < 0.0001). The correlation became weaker in the course of pregnancy, losing significance by week 32. This was explained by the differences in profiles of the two variables: the lowest measured APC ratio preceded the peak of factor VIII in most cases. The most pronounced rise of factor VIII was found in the women with minimal levels of APC ratio between 0.8 and 0.7. These results allowed us to speculate that APC response is closely regulated during pregnancy, aiming to maintain a certain relevant level. Transitory reduction of APC response is connected to factor VIII and discussed as a prevalent mechanism of functional APC resistance during pregnancy. PMID- 9199822 TI - Factor V antigen levels in APC resistance, in factor V deficiency and in combined APC resistance and factor V deficiency (pseudohomozygosis for APC resistance). AB - Factor V antigen levels were measured in 40 patients with factor V deficiency (11 homozygous and 29 heterozygous), in 38 patients with factor V Leiden mutation (16 homozygous and 22 heterozygous) and in three patients with combined heterozygous factor V deficiency and heterozygous factor V Leiden mutation (so-called pseudohomozygosis for APC resistance). Twenty normal subjects of both sexes served as controls. Factor V antigen levels compared well with factor V activity in normal subjects and in all groups of patients. They were normal both in homozygous and heterozygous APC resistance patients. Factor V antigen determination may be useful for the diagnosis of pseudohomozygosis for APC resistance. These patients have a phenotypic picture similar to homozygous APC resistance, but show a factor V antigen level about half the normal value since they are compound heterozygotes for factor V deficiency and APC resistance. In contrast, homozygous patients for APC resistance show normal factor V activity and antigen levels. PMID- 9199823 TI - Successful short-term oral surgery prophylaxis with rFVIIa in severe congenital factor VII deficiency. PMID- 9199824 TI - Incompatibility of a recombinant thromboplastin and an APTT reagent on a fully automated coagulation analyzer. PMID- 9199825 TI - Different contribution of factor V Leiden-associated hypercoagulability to ischaemic cerebrovascular disease in the elderly. PMID- 9199826 TI - Low plasma factor XII activity in patients with systemic lupus erythematosus and a lupus anticoagulant. PMID- 9199827 TI - Bacteriosclerosis. PMID- 9199828 TI - The genetics of glaucoma: an update. PMID- 9199829 TI - Retinal toxicity of intravitreous octreotide in the rabbit. AB - OBJECTIVE: To evaluate the ocular toxicity of intravitreous octreotide. DESIGN: New Zealand white rabbits weighing approximately 2 kg were given 5 mg (group 1, two eyes), 2 mg (group 2, four eyes), 1 mg (group 3, four eyes), 0.5 mg (group 4, two eyes), 0.3 mg (group 5, two eyes) or 0.1 mg (group 7, two eyes) of octreotide acetate, two doses of 0.3 mg 1 week apart (group 6, four eyes) or 0.1 mL of balanced salt solution (group 8 [control group], two eyes). OUTCOME MEASURES: Findings on clinical examination and electroretinography, performed before and 10 days after injection, and on light microscopy. RESULTS: Cataracts developed in groups 1 and 2. No clinical changes were found in groups 3 to 8. Electroretinography showed various degrees of decrease in the b-wave amplitude in groups 1 and 2; the results were normal in groups 3 to 8. Histologic examination showed macrophage and monocyte infiltration in the vitreous and retina in group 1. No histologic change was seen in the eyes in groups 2 to 8. CONCLUSIONS: Octreotide injected intravitreally is safe at dosages of 1 mg or less. PMID- 9199830 TI - Computerized videokeratography of keratoconus kindreds. AB - OBJECTIVE: To assess the role of heredity in the development of keratoconus. DESIGN: Prospective study. SETTING: Eye clinic providing secondary and tertiary ophthalmic care in Toronto. PATIENTS: Thirty-nine patients with keratoconus (57 eyes) and 48 relatives of 11 patients with keratoconus. The corneal topography of the family members was compared with that of a group of 68 volunteer control subjects (136 eyes) without clinical evidence or a family history of keratoconus. OUTCOME MEASURES: Three quantitative measures derived from computerized videokeratography: the relative steepness of the inferior cornea versus the superior cornea, central corneal power and the difference in central corneal power between the two eyes. All the data were statistically analysed with the use of nonparametric discriminant analysis. RESULTS: Fifteen family members who were believed to be clinically normal on the basis of refraction, keratometry and slit lamp examination has statistically significant topographic abnormalities suggestive of early or mild keratoconus. CONCLUSIONS: The presence of these findings in family members of patients with keratoconus may represent the incomplete expression of a gene contributing to the development of the condition. Pedigree analysis suggested an autosomal dominant inheritance pattern in 9 of the 11 families. Our results underline the value of videokeratography for accurate family pedigree analysis and the diagnosis of keratoconus. PMID- 9199831 TI - Modulation of the blood-aqueous barrier by light exposure in patients with uveitis. AB - OBJECTIVE: To evaluate the effect of light deprivation on flare measurements in patients with uveitis. DESIGN: Prospective study. SETTING: Eye clinic providing tertiary ophthalmic care in Calgary. PATIENTS: Five consecutive patients with a history of long-standing uveitis. INTERVENTIONS: Flare measurements were obtained with the Kowa FC-1000 flare-cell meter before and after 24 hours of monocular occlusion, and before and after wearing sunglasses during waking hours in a 24 hour period. OUTCOME MEASURE: Aqueous flare. RESULTS: A significant decrease in flare was observed after 24 hours of occlusion in all patients (p < 0.01). One patient showed an interesting change in response when her steroid treatment was tapered and subsequently stopped. Two of three patients showed a decrease in flare after wearing sunglasses. CONCLUSIONS: These preliminary results suggest that sunlight causes breakdown of the blood-ocular barrier in certain cases of uveitis. Further investigation is needed to determine the extent and mechanisms of light-induced breakdown of the blood-aqueous barrier as well as the role of sunglasses and anti-inflammatory medications as potential modulators of light induced inflammation. PMID- 9199833 TI - Anterior subtotal vitrectomy with fibre-optic illumination. PMID- 9199832 TI - A magnetic resonance imaging study of double elevator palsy. AB - OBJECTIVE: The pathophysiology of double elevator palsy is poorly understood. We assessed two patients with this condition using magnetic resonance imaging (MRI) to evaluate the appearance of the extraocular muscles. DESIGN: Cross-sectional study. SETTING: Radiology department of a university-affiliated hospital in London, Ont. PATIENTS: Two patients from a private ophthalmology practice who had undergone complete transpositions of the horizontal rectus muscles to treat hypotropia associated with double elevator palsy. INTERVENTION: MRI. A volume scanning technique was used to obtain maximum information about the muscles. OUTCOME MEASURE: Appearance of the extraocular muscles. RESULTS: In both patients MRI showed decreased volume of the superior rectus muscle on the affected side. The other rectus muscles were normal. This suggested either congenital hypoplasia or paresis of the involved superior rectus muscle. In addition, the full tendon transpositions of the medial and lateral recti did not appreciably change the middle and deep orbital pathways of the transposed horizontal rectus muscles. CONCLUSIONS: MRI may be a useful adjunct to saccadic velocity assessments in differentiating between primary inferior rectus restriction, primary superior rectus paresis and congenital supranuclear elevator deficiency. PMID- 9199834 TI - Some observations concerning visual perception during vitrectomy after retrobulbar anesthesia. PMID- 9199835 TI - Superior oblique myokymia as a bilateral subjective phenomenon. PMID- 9199836 TI - Angiocentric lymphoma presenting as massive unilateral eyelid swelling. PMID- 9199837 TI - Anterior segment recurrence of acute myelogenous leukemia: treatment with subconjunctival injections of methotrexate and triamcinolone acetonide. PMID- 9199838 TI - Neuroimaging techniques for the ophthalmologist. PMID- 9199839 TI - Polyhydramnios. PMID- 9199840 TI - Physiology of amniotic fluid volume regulation. AB - Although there are fairly wide variations AFV normally undergoes characteristic changes across gestation in which it increases from 10-20 ml at 10 weeks gestation to average 800 ml at 24 weeks. Little change occurs from then until near term when AFV begins to decrease, and large decreases can occur in postterm pregnancies. Across gestation, 95% of AFVs are within the range of 1/2.57-2.57 times the gestational mean volume and 99% are within the range of 1/3.40-3.40 times the gestational mean. Although there are six pathways in which fluid and solutes can enter and/or leave the amniotic sac, there are only four primary pathways that contribute to AFV during late gestation. These include fetal urine and lung fluid secretion as the two primary sources of fluid, with fetal swallowing and intramembranous absorption as the two primary routes of amniotic water clearance. The intramembranous pathway also appears to be a primary source of amniotic solutes (e.g., sodium and chloride). Although fetal hypoxia has been widely believed to cause oligohydramnios, fetal hypoxic hypoxia and anemic hypoxia both appear to be associated with an increased AFV and polyhydramnios rather than oligohydramnios. It is speculated that the oligohydramnios associated with fetal hypoxia is caused by placental dysfunction in addition to the hypoxia. PMID- 9199841 TI - Current thoughts on twin-twin transfusion syndrome. PMID- 9199842 TI - Clinical assessment of amniotic fluid. AB - Amniotic fluid volume estimation has become an integral part of fetal evaluation. Although the sonographic techniques clinically available are limited in their accuracy and predictive value, the careful performance of AFI measurements provide important and complementary clinical data on which to base management decisions in pregnancies at risk. PMID- 9199843 TI - Oligohydramnios: sonographic diagnosis and clinical implications. PMID- 9199844 TI - Therapeutic interventions for oligohydramnios: amnioinfusion and maternal hydration. PMID- 9199845 TI - Ultrasound measurement of fetal urine flow. AB - In summary, the measurement of fetal urinary flow remains controversial because the actual volumes of the human fetal bladder are unknown. The volume formula for an ovoid overpredicts fetal bladder volume and has a wide margin of certainty. The best ultrasonographic prediction of bladder volumes were calculated by applying a regression formula to the area of a sagittal sonographic image of the bladder (0.46323 + 1.39394.sagittal area). Calculations of the HFUFR based on the ovoid volume formula may be overestimated by 40%-70%. However, HFUFR estimated by regression of three to six longitudinal bladder area measurements is better with a narrow margin of uncertainty (+/- 35%). This technique affords improved accuracy and is easier and more convenient. PMID- 9199847 TI - Current status of amniotic fluid tests of fetal maturity. PMID- 9199846 TI - Fetal swallowing: relation to amniotic fluid regulation. AB - In summary, fetal swallowing activity contributes importantly to fetal and amniotic fluid homeostasis, and fetal somatic and gastrointestinal development. Human and ovine fetal swallowing increases throughout gestational with fetal swallowed volumes markedly greater (relative to body weight) than adults. Although the regulation of swallowing activity in early gestation is unknown, intact central and systemic dipsogenic mechanisms have been shown during the last third of ovine gestation. Recent studies suggest that swallowing behavior may be modulated in accordance with neurobehavioral state changes and influenced by hypoxia, hypotension and plasma osmolality changes. Whether fetal swallowing also is regulated by the development of "hunger" sensation, salt appetite, or the development of taste is uncertain. Nevertheless, it is likely that, for species in which swallowing behavior develops in utero, there are potentially dramatic influences of the maternal-fetal pregnancy environment on the imprinting of regulatory mechanisms controlling ingestive behavior. Ultimately, the regulation of fetal swallowing may aid in the prevention and/or therapy of human amniotic fluid disorders. PMID- 9199848 TI - The spectrum of managed care model types. AB - The health care delivery landscape is becoming increasingly complicated. Delivery systems and insurance products are attempting to balance choice with value. Large purchasers are deciding between taking on risk and designing their own benefit package as opposed to paying for the insurance companies to assume their risk and accepting the federally qualified benefit package. PMID- 9199849 TI - Looking down the prospectoscope: obstetrics/gynecology in the year 2000 and beyond. AB - It should be emphasized that the American College of Obstetricians and Gynecologists is advocating that those Obstetrician/Gynecologists who wish to be included under this new designation primary care physician "provide only ambulatory primary care," states the Executive Director, Dr. Ralph W. Hale. Thus, we enter a new era and must understand the lexicons, including HMO, independent provider organization, PPO, managed care organization, exclusive provider organization, IPA, hospital-physician organization, health plan provider data and information set, physician-hospital organization and utilization management--all these have become words with increased meaning for all clinicians. It is a rare physician lounge without physicians preoccupied by discussions regarding managed care, "Have you heard the latest reimbursement schedule?" or "It's less than Medicaid," and the discussions continue indefinitely. There has been a continued effort to have physicians be cost-efficient in their approach to all aspects of obstetrical and gynecological care. Capitation has proven to be the virtually exclusive method of reimbursement. Continued care with respect to clinical outcome, resource utilization, patient satisfaction, and quality care restructuring of practice/personnel/patient approach is based on the quarterly patient surveys, which have been evaluated and carefully reviewed. Patient satisfaction with a sincere effort to provide quality of care remains the underlying theme with respect to obstetrical and gynecological patient care. These are basic tenets. What is the future of obstetrics/gynecology with respect to managed health care? It does make sense to have a planned and well-coordinated approach to delivery of obstetrical as well as gynecological care with the goal of "quality" delivered at a "lower cost" having an overall positive impact on OB/GYN health care delivery. Currently, there are a number of states which have specific legislation enabling a patient to proceed to secure gynecological without going through a "gatekeeper." As we approach the year 2000 and beyond, clearly the prediction is that this will be an ever-increasing goal and objective of state medical societies to provide easy access for women's health care. Thus, we have awakened into a new era that is hallmarked by efficient, quality medical care provided by a physician who initially trained to be a subspecialist and now is an ambulatory primary care physician. PMID- 9199850 TI - Obstetrician/gynecologist as primary care physician in managed health care. PMID- 9199851 TI - Managed obstetrical care. AB - The current maternal/newborn care model is outdated and needs to be revised. The health care reform movement has created a window of opportunity to redefine the episode of pregnancy care and develop a more meaningful and more cost-effective model of care. The ultimate satisfaction for physicians will occur when they exert their natural control regarding the manner in which health care dollars are spent by managing the financial risk and the patient care. The optimal management of the health care dollar can only be achieved through initiation of an integrated model in which a coordinated care team supported by the appropriate risk assessment, education, prevention and wellness program, and medically necessary intensive care of the high-risk pregnancy are brought together effectively. An integrated model will give patients what they want: compassionate, convenient, comprehensive care. It will give the payers what they are looking for--appropriate care at an appropriate predicable cost and improved outcomes. Finally, it will give the providers what they want: control over the delivery medical care. PMID- 9199852 TI - Managed health care's approach to infertility. PMID- 9199853 TI - Preparing obstetricians and gynecologists for capitation. AB - We are indeed facing a changing paradigm in the way medical services are delivered and reimbursed. Physicians are becoming entangled in the business world. Terms such as "medical loss ratio" must be integrated within the physician's existing language. According to Green and Barnett, "No group is so large or so small as to likely survive the next decade without being involved in a capitated contract." The change is not tomorrow, it is today. Do not be lost in the changing world of medicine without becoming involved, and see if it is for you and your practice. Progress of medicine may not be what you would like but do not be passed by without making an impact. Capitation offers an opportunity to regain control of the management of care, to develop innovative and cost effective approaches to delivery, and to enhance income and revenue in the face of declining health care budgets. PMID- 9199854 TI - A performance-based compensation model for obstetricians/gynecologists. PMID- 9199855 TI - Obstetrics/gynecology network development. PMID- 9199856 TI - Anger and hostility in maritally violent men: conceptual distinctions, measurement issues, and literature review. AB - Marital violence researchers have generally used the terms anger and hostility interchangeably. However, there are important differences between anger and hostility that may be vital to understanding the relationship between these constructs and marital violence. The present manuscript highlights the advantages of distinguishing between anger and hostility. In order to investigate the role of anger and hostility in marital violence, we provide a comprehensive review of 26 empirical studies in addition to critically examining researchers' definitions of anger and hostility and the methods of assessment utilized in this body of research. While many researchers have presented data suggesting that maritally violent men are higher in anger and hostility than maritally nonviolent men, the findings are not consistent and vary in accordance with the construct assessed and the assessment strategy used. PMID- 9199857 TI - Psychological factors affecting health after toxicological disasters. AB - Exposure to toxic substances in the environment is an ever more common event, that may cause physical as well as psychological harm. When an entire community is exposed, the term 'toxicological disaster' is used. The mere threat of such an event may be a source of stress, associated with changes in mental health, physical health, and changes in health-related behaviors. A review is presented of the literature about the effects of the stressful experience of toxicological disasters on health and health-related behaviors. Three questions are examined: (a) do toxicological disasters represent a specific type of stressor, different from other stressors?; (b) which stress-mediated health effects have been observed in the aftermath of toxicological disasters? and (c) is there evidence for a higher vulnerability in certain identifiable risk groups? On the basis of the available literature, it is concluded that toxicological disasters may have profound effects on subjective health, especially on symptom reporting, and on a number of psychophysiological parameters. Evidence for a substantial impact of disaster-related stress on either physical or psychiatric morbidity remains inconclusive. In this respect toxicological disasters do not appear to differ from other stressors. There is some evidence that toxicological disasters may have a more pronounced effect on health-related behaviors, especially on reproductive behavior (number of births and abortions). Women, and especially those who have young children to care for, appear to be more at risk for the observed health effects. The evidence for a higher vulnerability in other risk groups (e.g., former psychiatric patients remains inconclusive. PMID- 9199859 TI - An information-processing perspective on childhood anxiety. AB - In the past decade, cognitive theories of adult anxiety disorders have become increasingly complex, reflecting enhanced understanding of anxiety-related information-processing. This growth has fostered the development and enhancement of numerous assessment and treatment methods. Unfortunately, similar growth has been slower to occur in theories of childhood anxiety. This paper attempts to foster such growth by adopting an information-processing perspective. Doing so expands the extant cognitive perspective on childhood anxiety in four major ways. First, the division of cognitive processing into a sequence of steps provides a framework for organizing predictions regarding cognitive factors in childhood anxiety. Second, consideration of the cognitive operations active during each stage in the sequence facilitates elaboration of the types of cognitive deficits and distortions characteristic of anxious children. Third, it promotes development and application of performance-based assessment methodologies. Finally, an information-processing perspective highlights several targets for clinical intervention that may promote widespread change in an anxiety-supporting cognitive system. PMID- 9199858 TI - Contributions of risk-factor research to developmental psychopathology. AB - Risk-factor research refers to the study of antecedent conditions and subsequent outcomes and the ways in which these are interrelated. The research encompasses a broad range of questions and research strategies. The paper discusses the characteristics and contributions of risk-factor research in the context of developmental psychopathology. The ways in which causal paths are conceptualized, the capacity to integrate multiple influences, and applications that can be derived from the findings are discussed. The progression of research is illustrated in relation to key concepts (correlate, risk factor, marker, causal risk factor) that reflect varied levels of understanding antecedent-outcome relations. The identification of causal relations, progressions, and paths over the course of development and the interplay of theory, research, and application are illustrated and discussed. PMID- 9199860 TI - Children's nighttime fears. AB - Some children experience persistent night-time fears that interfere with their daily functioning. Initially, we present developmental considerations necessary to an understanding of severe night-time fears. We postulate that severe night time fears are probably due to a complex interaction of biological, environmental, and cognitive-mediational processes. Several assessment procedures are outlined: behavioral interviews, diagnostic interviews, fear survey schedules for children, home monitoring on the part of parents, and darkness toleration tests. Traditional behavioral interventions, and more recent cognitive-behavioral interventions, are evaluated in terms of their research foundations. Cognitive behavioral strategies appear to have the more empirical support, although we draw attention to several methodological limitations. PMID- 9199861 TI - Daily functioning of the lower extremity amputee: an overview of the literature. AB - The aim of this paper is to review the existing literature on the incidence, morbidity and mortality of lower limb amputation. The functional level of the lower limb amputee and the predictive factors for functioning with a prosthesis are reviewed, both for unilateral and for bilateral amputees. The reported incidence of lower extremity amputation (LEA) varies considerably between different Western countries. The mean survival of LEA patients ranges between two and five years. Assessment of functional outcome is carried out differently. Studies are not comparable and most concern selected groups of amputees. Increasing age, concurrent disease and poor compliance are prognostic factors for a low functional level. For optimal planning of rehabilitation it is necessary to study a complete cohort of amputees with respect to these prognostic factors. PMID- 9199862 TI - A randomized controlled trial of enhanced Social Service occupational therapy for stroke patients. AB - OBJECTIVE: To determine whether stroke patients referred to the Social Service occupational therapy service would benefit from an enhanced service compared to the usual service. DESIGN: Randomized controlled study allocating patients to the enhanced service or the usual service. SUBJECTS: Stroke patients discharged home from hospital and referred to Social Service occupational therapy department. OUTCOME MEASURES: The sections and total score from the Nottingham Extended Activities of Daily Living Scale (EADL), the Barthel Index, the General Health Questionnaire (GHQ) and the number of pieces of equipment provided were analysed. RESULTS: One hundred and eleven stroke patients were recruited to this study. Fifty-three were randomly allocated to the enhanced service and 58 to the usual service. Patients receiving the enhanced service were seen more quickly after referral, for longer, and received significantly more visits (p < 0.01) than those receiving the usual service. Three months after entry to the study the enhanced service group had better EADL (p < 0.01) than the usual service group. This benefit remained significant in only the mobility section of the EADL at six months. Careers of the stroke patients in the enhanced group had lower GHQ scores (p < 0.05) than those in the usual group at six months. CONCLUSIONS: This trial supports the use of domiciliary occupational therapy for stroke patients after discharge from hospital in terms of improvements in functional outcomes in the short term, but the long-term benefits remain unclear. PMID- 9199863 TI - Evaluation of cognitive behavioural treatment for depression after stroke: a pilot study. AB - OBJECTIVE: To evaluate cognitive behavioural therapy in the treatment of depressed stroke patients. DESIGN: A series of AB single case experimental design studies. SUBJECTS: Stroke patients who had been admitted to hospital. METHODS: Patients were identified from a hospital register of stroke patients and sent the Beck Depression Inventory (BDI) and Hospital Anxiety and Depression Scale to determine whether they were depressed. Those who were depressed were sent the BDI weekly during a four-week baseline period. They were then treated and continued to complete the BDI weekly over a three-month treatment period. Patients were also assessed pre- and post-treatment on measures of functional independence. INTERVENTION: Cognitive behaviour therapy. RESULTS: Of the 626 patients on the Stroke Register, 373 returned questionnaires and 155 were identified as depressed. Of these 136 were visited and 19 agreed to take part. The outcome was evaluated using three methods: visual inspection, comparison of patients during baseline and treatment phases and proportion of scores below lowest baseline. This showed that four patients consistently showed beneficial treatment effects, six patients showed some benefit and nine no benefit. For the group as a whole there was a significant decrease in depression during the treatment period on the BDI (p = 0.02) but no significant change in functional abilities. CONCLUSION: Cognitive behavior therapy is an appropriate treatment for some depressed stroke patients and beneficial for some patients. Further evaluation of this treatment with stroke patients is warranted. PMID- 9199864 TI - Effects of electrical stimulation on flexion contractures in the hemiplegic wrist. AB - OBJECTIVE: To study the effects of electrical stimulation (ES) on flexion contractures in the hemiplegic wrist. DESIGN: The investigation was carried out following an OFF (two weeks with rehabilitation only)--ON (two weeks with ES treatment and rehabilitation)--OFF (two weeks rehabilitation only) fixed protocol. SETTING: A stroke ward and an outpatient stroke service. SUBJECTS: Eleven hemiplegic subjects with reduced range of extension and increased resistance to passive movement at the wrist. MAIN MEASURE: Quantitative measures of the hemiplegic posture at the wrist, passive range of extension and resistance to passive extension of the wrist. Measurements were taken at the start of the study and then at two-weekly intervals. Two extra measurements were taken at the end of the ON period. RESULTS: Following two weeks treatment with ES the posture of the wrist improved and the passive range of extension increased. However, there were no significant changes in the resistance to passive movement. These benefits appeared largely to be lost two weeks after ES was discontinued. CONCLUSIONS: Short-term ES gives temporary improvements in contractures at the wrist in poststroke hemiplegia. PMID- 9199865 TI - Comparison of postal version of the Frenchay Activities Index with interviewer administered version for use in people with stroke. AB - OBJECTIVE: To assess the agreement between postal and interviewer-administered versions of the Frenchay Activities Index (FAI) and to assess the criterion validity of the postal version, using interviewer administration as a gold standard. DESIGN: Comparison of responses to FAI administered by post and then by interview (median delay 10 days). SUBJECTS: Forty-eight Oxfordshire residents admitted to hospital with acute stroke between 1 August 1994 and 31 January 1995 and discharged alive within six months of their stroke. RESULTS: The limits of agreement of the total FAI score are from -5.4 to 7.2. The kappa statistic for each of the 15 individual items that make up the FAI ranged from 0.35 to 1. For nine items, agreement was moderate or fair, and for six items, agreement was good or very good. The mean difference between the overall scores was 0.9 (95% confidence interval: -0.1 to 1.9). The correlation between the overall scores was 0.94 (Spearman's rank correlation coefficient). CONCLUSION: The postal version of the FAI is a satisfactory alternative to direct administration, but poor agreement in scores for individual patients emphasizes that the two approaches should not be used sequentially to monitor individual patients. PMID- 9199866 TI - Subjective well-being one year after stroke. AB - OBJECTIVE: To compare the subjective well-being of stroke patients with that of a reference group, and to study its relationship to patient characteristics. DESIGN: Cross-sectional study. SETTING: Interviews performed in the respondents' homes, tests performed at the outpatient clinic. SUBJECTS: Sixty patients one year after stroke (median age 74 years, interquartile range (IQR) 68-80), and 419 reference individuals (median age 75 years, IQR 71-80). MEASURES: Subjective well being assessed with the General Health Questionnaire (GHQ-20). Explanatory variables were demographic and medical characteristics of the individuals and scores on validated tests: Barthel Index, Frenchay Activities Index (FAI), Sodring Motor Evaluation of Stroke Patients, Assessment of Cerebral Stroke and other Brain Damage, and Mini-Mental State Examination (MMSE). RESULTS: A significantly higher proportion of the stroke patients than of the controls rated their subjective well-being as low, also after adjustment for age and gender (adjusted odds ratio 20.1, 95% confidence interval 9.6-42.0 by logistic regression). In bivariate analyses, leg and arm motor impairment, visuospatial impairment, apraxia, aphasia, low Barthel score, low FAI score, low MMSE score, and institutionalization were highly significant predictors of low subjective well-being (p-values < 0.01). In multiple linear regression, a model with gender (p = 0.3) and upper extremity motor score (p < 0.01) fitted the data well, and explained 48% of the variance in GHQ. CONCLUSION: Subjective well-being is decreased one year after stroke, and this is mainly attributed to arm motor impairments. PMID- 9199867 TI - Development of a neurological rehabilitation environment: an observational study. AB - BACKGROUND: Engagement in therapeutic activity among stroke inpatients is advocated by many rehabilitation professionals. However, there is a lack of published evidence to indicate whether this is currently being achieved. OBJECTIVE: To investigate the extent and types of 'rehabilitation' activities on a new neurological rehabilitation ward, and examine change in patients' behaviour related to the new environment and new initiatives. DESIGN: Five one-week observation periods were conducted over two years, with a total of 67 patients being observed. A comparison was made with results of an earlier study of stroke patients' activities conducted at the same hospital. RESULTS: Patients spent an average 43 min per day with therapists (9% of the working day) and this was only marginally supplemented by self-exercise (2% of the working day--approximately 10 min). However, the provision of a new rehabilitation environment was associated with a marked decrease in the time patients spent at their bedsides, and a decrease in time spent passively gazing or watching others. CONCLUSIONS: Overall there was some positive change in patients' behaviour. We suggest that structured guidance is required from the whole multidisciplinary team to stimulate more self directed therapy practice and recreation. PMID- 9199868 TI - Setting handicap goals with elderly people: a pilot study of the Life Strengths Interview. AB - OBJECTIVE: To assess whether the Life Strengths Interview (LSI) is a useful clinical framework to identify handicap goals. DESIGN: Clinical case studies. SETTINGS: Two elderly care rehabilitation hospitals. SUBJECTS: Five people, whose ages ranged from 73 to 90 years. All participants were aware of their likely resultant disability, scored 25+ out of a possible 30 with the Mini-Mental State Examination, were able to communicate effectively and were due to be discharged home in approximately one month. INTERVENTIONS: Each participant undertook the LSI process with the research occupational therapist. MAIN OUTCOME MEASURES: Identified rehabilitation goals and their achievement. RESULTS: Goals were focused around families and other support networks. Six to eight weeks following discharge, achievement of goals varied. CONCLUSIONS: This pilot study suggests that the LSI may be a useful clinical framework but further research needs to investigate whether a modified clinical version may be more suitable. PMID- 9199869 TI - Abnormal illness behaviour in rehabilitation from stroke. AB - OBJECTIVE: To examine the effect of abnormal illness behaviour (AIB) on rehabilitation outcome following stroke. DESIGN: A longitudinal design, with assessments on admission to and discharge from rehabilitation, and six and 12 months after discharge. SETTING: The study was undertaken in the rehabilitation unit at Repatriation General Hospital, in Adelaide, South Australia. SUBJECTS: Ninety-four 12-month stroke survivors who had undergone an inpatient rehabilitation programme. MAIN OUTCOME MEASURES: AIB was assessed using the Illness Behaviour Questionnaire. Additional psychological measurements comprised the Zung Self-Rating Depression Scale, General Health Questionnaire, and a visual analogue mood scale. Functional ability was assessed with the Australian ADL Index, and lifestyle activities with the Frenchay Activities Index. RESULTS: Cluster analysis of discharge data was used to define a rule for identifying patients with AIB. AIB was apparent in nearly 30% of patients at discharge, and persisted for 12 months. Patients with AIB scored more poorly than non-AIB patients on functional, social and psychological indicators. CONCLUSIONS: AIB emerged as a key determinant of long-term disability. It is important to consider why AIB develops during rehabilitation, and how to identify patients at risk. PMID- 9199870 TI - Are there gender differences in functional outcome after stroke? AB - PURPOSE: To study gender differences in functional outcome unexpectedly observed in a follow-up study of stroke patients. DESIGN: Prospective study of hospitalized stroke patients, with evaluations in the subacute phase and after one year. SETTING: Geriatric and general medical wards, and geriatric outpatient clinic of a university hospital serving as general hospital for a defined population. SUBJECTS: All stroke patients admitted during a six-month period (n = 165) were considered for inclusion, of whom 87 could be assessed in the subacute phase and 65 after one year. MAIN OUTCOME MEASURES: Motor function assessed by the Sodring Motor Evaluation of Stroke Patients; cognitive function by the Assessment of Stroke and other Brain Damage; and activities of daily living (ADL) function by the Barthel Index. Nursing-home residency registered after one year. RESULTS: Men achieved a significantly better score than women on most of the scales used. The age-adjusted odds for a man to have a higher Barthel score than a woman was 3.1 (95% confidence interval (CI) 1.3-7.0) in the subacute phase and 3.3 (95% CI 1.2-9.0) after one year. Differences of the same magnitude were seen on the subscales of the motor and cognitive tests. The same trend was observed on all items of the Barthel Index. The males had a lower likelihood to be permanent nursing-home residents after one year, the age-adjusted odds ratio for nursing home residency for females versus males being 6.3 (95% CI 1.2-65.3). CONCLUSION: Women seem to be functionally more impaired by stroke than men. PMID- 9199871 TI - Murine lymphotactin: gene structure, post-translational modification and inhibition of expression by CD28 costimulation. AB - The production of lymphokines by T cells is dependent on TCR signalling and is enhanced by costimulatory signals through CD28. In IL-2 producing T cells clones (Th1 cells), TCR signalling in the absence of costimulatory signals renders these cells unable to product IL-2 in response to subsequent stimulation through the TCR and CD28. Using a differential screening approach we independently cloned murine lymphotactin from a cDNA library produced from an unresponsive Th1 cell. Resting Th1 clones and freshly isolated CD8+ T cells produced lymphotactin mRNA maximally in response to TCR stimulation alone and expression was surprisingly inhibited by CD28 costimulation, at least early after activation. In addition, Th1 cells made unresponsive by prior TCR stimulation, produced lymphotactin but not IL-2 mRNA when restimulated through the TCR and CD28. Sequence analysis of the lymphotactin gene and mapping of the transcription initiation sites have delineated the promoter region and the boundaries of exons and introns. Studies on post-translational modification of lymphotactin demonstrate the cleavage of the signal peptide containing the first two cysteine residues in the predicted amino acid sequence and suggests that the secreted lymphotactin is an O glycosylated protein. PMID- 9199872 TI - Susceptibility to virus infection is determined by a Stat-mediated response to the autocrine effect of virus-induced type I interferon. AB - Interferon (IFN) signalling appears to be the predominant pathway that leads to the development of the host defence against virus. However, other mechanisms that are IFN independent may also be involved. The Stat1 transcription factor is specific for the IFN pathway and plays a central role in mediating many, if not all, IFN-dependent biological responses. Viral infection of cells in the presence or absence of Stat1 was studied to examine the role of IFN-dependent and independent antiviral mechanisms. A lower virus yield measured by plaque assays was observed following challenge with VSV, EMCV and NDV of cells in which either Stat1a or Stat1b was present compared to those lacking both forms of the Stat protein. The more efficient antiviral response was abolished when cells were infected with virus in the presence of IFN-neutralizing antibodies, suggesting that a Stat-dependent pathway is activated following virus infection by endogenously produced IFN. Virus-induced Stat protein translocation from the cytoplasmic compartment, detected within 3 h of infection, and Stat-dependent transcriptional activation of ISGF2 in response to virus challenge, were also abolished by neutralizing type I IFN. Thus, susceptibility to virus infection can be determined by the cell's ability to respond to autocrine IFN through the Stat mediated pathway of gene induction. PMID- 9199873 TI - Regulation of Fas and Fas-ligand expression in NK cells by cytokines and the involvement of Fas-ligand in NK/LAK cell-mediated cytotoxicity. AB - This study demonstrates cytokine-mediated regulation of Fas and Fas-ligand (Fas L) expression in human NK cells and the involvement of the Fas-L pathway in NK/LAK cytotoxicity. Freshly isolated, high purified human CD56+CD3- NK cells were found to express Fas and Fas-L. Cytokines further increased the Fas expression in the CD56+CD3- NK cells, with interleukin (IL)-2 being the most potent stimulus followed by IL-12, while IL-7 had no effect. IL-2 and IL-7 equally enhanced the Fas expression in the CD56+CD3+ population, while IL-12 had a less pronounced effect. Incubation of the CD56+CD3- NK cells with IL-2, but not with IL-12 and IL-7, led to an upregulation at the Fas-L expression, whereas neither of the cytokines affected the Fas-L expression in the CD56+CD3+ cells. Antagonistic Fas mAb M3 and Fas-IgG1 fusion protein significantly inhibited NK cytotoxicity towards Fas-expressing Jurkat cells, while non-antagonistic Fas mAb M31 and irrelevant CD14-IgG1 fusion protein had no effect. IL-2-generated LAK cells were much more potent than NK cells in exerting the cytotoxic effect on Jurkat cells, which was also partially inhibited to M3 and Fas-IgG1. Thus, human NK and LAK cells express Fas and Fas-L, utilize the Fas-L cytotoxic pathway and enhance the expression of these molecules in response to cytokines. PMID- 9199874 TI - Cytokine mRNA expression in human platelets and a megakaryocytic cell line and cytokine modulation of platelet function. AB - Platelet formation and function are regulated, in vivo, to varying degrees by cytokines in the micro-environment. While white blood cells are the major source of cytokines within the cardiovascular system, the question addressed in this study was whether platelets and the platelet precursor, the megakaryocyte, may also serve as a source of cytokines. Cytokines produced by or carried within platelets could be released at sites of vascular injury and participate in wound healing. Platelets and a human megakaryocyte-like cell line, HU3, were found to express message for interleukin 7 (IL-7), stem cell factor (SCF), transforming growth factor beta (TGF-beta), cMpl, the IgE receptor subunits Fc epsilon RI alpha gamma and the transcription factor, NF-E2. Other cytokines expressed in HU3 cells but not in platelets included IL-1 beta, IL-6, IL-10, IL-13, TNF-alpha and the FC epsilon RI beta subunit. The HU3 cell line seemed to be further along the maturation/differentiation pathway to platelet formation than a second blood derived bipotential cell line, MB02. The MB02 cell line did not express IL-6, IL 10, SCF, TNF-omega nor cMpl. Furthermore, culturing the HU3 cells in TPO appeared to repress expression of Fc epsilon RI beta directing the cell closer to the platelet phenotype. In light of the presence of cytokine expression in platelets/megakaryocytes, agonist-induced platelet aggregation was measured in the presence of added cytokines as a means to evaluate potential cytokine modulation of platelet function. Collagen-induced aggregations were significantly enhanced by IL-6, SCF and TPO. Other cytokines tested significantly stimulated the thrombin receptor activating peptide, SFLLRNP-, U46619- and ADP-induced platelet aggregations with TPO being the most consistent activator. It is possible that cytokines released from platelets act in concert with cytokines released from other cellular sources to modulate haemostasis and thrombosis differentially depending upon the site of injury. PMID- 9199875 TI - Induction of gelatinase B and MCP-2 in baboons during sublethal and lethal bacteraemia. AB - Intravenous injection of sublethal or lethal doses of Escherichia coli in baboons resulted in increased serum levels of the matrix metalloprotease gelatinase B and the chemokine monocyte chemotactic protein 2 (MCP-2). In both animal models, gelatinase B appeared faster than MCP-2. After sublethal challenge, serum levels of gelatinase B and MCP-2 were found to be correlated, reaching peak levels between 2 and 4 h after bacterial challenge. After lethal challenge, however, MCP 2 tended to increase until 10 h. The kinetics of appearance suggest induction of release of gelatinase B and de novo synthesis and secretion of MCP-2, both by endotoxin. PMID- 9199876 TI - T cell subsets and cytokines in allergic and non-allergic children. I. Analysis of IL-4, IFN-gamma and IL-13 mRNA expression and protein production. AB - Interleukin 4 (IL-4) and IL-13 are key cytokines inducing switching to immunoglobulin E (IgE), whereas interferon gamma (IFN-gamma) acts inhibitory on this process. We analysed whether differences existed in IL-4, IFN-gamma and IL 13 mRNA expression and protein production between T cells of children with allergic and non-allergic asthma, atopic dermatitis and health control children. IL-4 mRNA expression was increased in stimulated T cells of children with allergic asthma and atopic dermatitis, but not in those with non-allergic asthma as compared with healthy controls. Thus the increase in IL-4 expression can be considered as an underlying mechanism of the allergic disease process and not so much of the asthmatic state of the children. In unstimulated T cells of children with atopic dermatitis increased IFN-gamma mRNA expression with a reduced IFN gamma protein production was found, indicating a post-translational defect in IFN gamma. Differences in IL-13 expression between the groups were not significant, but IL-13 was significantly correlated with the height of the radio-allergo sorbent test (RAST) class and with the severity scoring of atopic dermatitis (SCORAD) index. This indicates the clinical relevance of IL-13 for the degree of allergen-specific sensitization and severity of atopic dermatitis. In conclusion, the imbalance in IL-4 and IFN-gamma secretion in patients with atopic dermatitis may reflect general T cell activation in the presence of an intrinsic defect of IFN-gamma secretion. PMID- 9199877 TI - T cells subsets and cytokines in allergic and non-allergic children. II. Analysis and IL-5 and IL-10 mRNA expression and protein production. AB - Interleukin 5 (IL-5) has an enhancing effect on IL-4 induced immunoglobulin E (IgE) synthesis. Furthermore, IL-5 plays an important role in the differentiation, recruitment, activation and survival of eosinophils. IL-10 has a downmodulating effect on interferon gamma (IFN-gamma) production and can exert strong anti-inflammatory activities. Therefore, we analysed whether differences were present in IL-5 and IL-10 mRNA expression and protein production between T cells of children with allergic and non-allergic asthma, atopic dermatitis and healthy control children. We demonstrated significant increases in IL-5 mRNA expression and protein production in different T cell fractions of children with allergic and non-allergic asthma and children with atopic dermatitis as compared to healthy controls. This indicates that IL-5 is not only involved in allergy, but also plays a role in the inflammatory process of non-allergic asthma. Interestingly, IL-10 mRNA expression by purified T cells of children with allergic and non-allergic asthma and children with atopic dermatitis was strongly decreased as compared with that of healthy controls. In the peripheral blood mononuclear cell (PBMC) fraction, IL-10 mRNA expression was comparable between the four groups. We hypothesize that this decreased T cell derived IL-10 expression results in a lack of immunosuppression of the inflammatory process in these diseases. However, a role of monocyte derived IL-10 cannot be ruled out. PMID- 9199878 TI - Effects of prolactin and metoclopramide on macrophage cytokine gene expression in late sepsis. AB - Previous studies indicate a profound suppression of tumour necrosis factor alpha (TNF-gamma), IL-1 beta and IL-6 release capacity by peritoneal macrophage (PM phi), splenic macrophage (SM phi) and Kupffer cells (KC) during late sepsis. Such a loss of functional capacity may reduce the animal's ability to ward off infection. Prolactin is known to enhance monocyte, T- and B-lymphocyte immune responses under normal conditions and has beneficial effects on cell-mediated immunity after haemorrhage. In the respect, the dopamine antagonist, metoclopramide, has been reported to increase circulating prolactin levels. Nonetheless, it remains unknown whether prolactin or metoclopramide have any salutary effect on macrophage (M phi) cytokine gene expression following sepsis. To study this, male C3H/HeN mice were subjected to sepsis and immediately thereafter were treated with prolactin (100 micrograms/25 g body weight, s.c.), metoclopramide (100 micrograms/100 g BW, s.c.) or given saline. PM phi, SM phi and KC (only SM phi and KC in metoclopramide-treated animals) were isolated at 24 h after sepsis. The monolayers were stimulated with or without LPS 10 micrograms/ml for 1 h in vitro. Total RNA was extracted and mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). A significant depression of constitutive and inducible mRNA levels of IL-1 beta, IL 6 and TNF-alpha in all three M phi populations were observed, when compared with shams (with exception of KC IL-6 mRNA in unstimulated cells). Prolactin as well as metoclopramide treatment after the onset of sepsis caused significant elevation of constitutive and inducible cytokine gene expression in all macrophages examined. Thus, prolactin and metoclopramide enhance the depressed M phi gene expression and may be useful in improving cell-mediated immunity during sepsis. PMID- 9199879 TI - IL-6 and soluble IL-6 receptor levels change differently after surgery both in the blood and in the operative field. AB - To investigate alterations in post-operative levels of IL-6 and soluble IL-6 receptor (sIL-6R), we examined their levels in serum and samples of drainage fluids from 26 patients who underwent thoracoabdominal surgery. Serum IL-6 levels reached the maximum within the first post-operative day and decreased thereafter. The IL-6 levels in the drainage fluid were much higher than in the serum (458 +/- 101-fold; mean +/- SEM) in the early post-operative phase. A large quantity of sIL-6R levels was present in blood samples. The time course of serum sIL-6R levels in 26 patients showed no significant change. sIL-6R concentrations in the drainage fluid were significantly lower than in serum (4.5 +/- 1.1-fold; mean +/- SEM) in the early post-operative phase. We propose that IL-6 is produced in the operative field and enters the peripheral blood stream to induce elevation of serum IL-6. On the other hand, sIL-6R levels in the operative field are lower than in the serum, and the serum sIL-6R levels are not influenced by surgical trauma. These data suggest that sIL-6R is being constantly produced in areas other than the operative field, while sIL-6R level is reduced by consumption in the operative field. Mechanisms to cope with surgical stress, involving sIL-6R together with its ligand IL-6 may thus exist. PMID- 9199880 TI - Increased susceptibility of Alzheimer's disease temporal cortex to oxygen free radical-mediated processes. AB - Reactive oxygen-mediated processes are though to contribute to the pathogenesis of Alzheimer's disease (AD). To investigate this hypothesis we studied autopsy tissue from 11 pairs of AD cases and control individuals matched for age, postmortem delay, and tissue storage time. The temporal neocortex, which is severely involved by AD pathology, and the cerebellum, which is spared, were analyzed for tissue markers of lipid peroxidation (LPO). The average chemiluminescence formed from bond breakage in tissue homogenates during a 3-h incubation, without the presence of catalysts such as metal ions or ascorbate, was significantly increased in the AD temporal cortex to 130% of matched controls. Basal tissue content of LPO products (thiobarbituric acid reactive substances--TBARs) was not different between groups. However, TBARs were significantly elevated in AD temporal cortex to 135% of control after the incubation. In contrast, in the cerebellum there was no difference between AD and control tissue, indicating a disease-specific tissue effect. Because the use of oral antioxidants have received considerable attention in the last few years, the results seen in the testing of an AD patient who took daily vitamin E supplements for 4 years is particularly interesting. The time course for CL reactivity in the temporal cortex was considerably delayed compared to all other samples. This observation is consistent with the hypothesis that antioxidants within tissue will quench ROS-mediated reactions. This study indicates that there is increased susceptibility to ROS in the AD temporal cortex that may contribute to the pathogenesis of the disease. Furthermore, our observation suggest that oral antioxidant supplementation may be protective against LPO in the human brain. PMID- 9199881 TI - Aging and caloric restriction affect mitochondrial respiration and lipid membrane status: an electron paramagnetic resonance investigation. AB - Previous studies have indicated that reactive oxygen species (ROS) are likely involved in the pathogenesis of neurodegenerative diseases including Alzheimer's disease (AD). ROS, generated by succinate-stimulated mitochondria, have been reported to be spin trapped and detected by electron paramagnetic resonance (EPR). Our aim in the current study was to study the impact of aging on the effect of increased metabolic stimuli on mitochondrial respiration in terms of oxy-radical generation and possible lipid peroxidative changes in brain neocortical membranes. A mixed population of brain synaptosomes and mitochondria from brown norway male rats of differing ages being fed either ad lib (AL) or a caloric-restricted diet (DR) was prepared and labeled with 5-nitroxyl stearate (5 NS), a membrane lipid-specific spin label. The changes in anisotropic motion of the intercalated 5-NS spin probe also allows one to evaluate the status of the membrane fluidity in the lipid microenvironment via the order parameter. Upon succinate stimulation of mitochondria, the ROS generated resulted in a decrease in the EPR signal amplitude of the 5-NS reporter molecule indicative of the flux of oxy-radicals produced and possible peroxidation-induced changes in the synaptosomal lipid membrane. The line width remained constant, indicating that the overall intensity was reduced. The results showed a significant overall age effect in the ability to generate oxygen-derived radicals following metabolic stimulation (p < .0001). Stimulation of state 4 mitochondrial respiration with 20 mM succinate resulted in greater oxy-radical production in 25-month-old animals as compared to younger animals, suggesting increased mitochondrial leakage with age. Free radical stress induced by metabolic stimulation also causes a concomitant increase in membrane fluidity (p < .0001). There was also a significant age effect (p < .0007) on the order parameter of the mixed population of membranes. Although caloric restriction attenuated the membrane rigidization caused by aging, it was found to play a role in limiting the oxy-radical production following metabolic stimulation of mitochondria. The overall effect of age on membrane spin-label intensities EPR signal upon succinate stimulation suggests that progressive mitochondrial dysfunction may be a key factor in the aging process and in development of age-associated diseases. PMID- 9199882 TI - Characterization of cholesterol oxidation products formed by oxidative modification of low density lipoprotein. AB - Oxidative modification of LDL is evidenced by alterations in both the protein and lipid components of the particle. Progressive oxidation of the apoprotein is associated with loss of specific amino acids and a gradual increase in electronegativity. Electronegative LDL has been isolated from human plasma (LDL-) by several groups using liquid chromatographic techniques and appears to be oxidized based on increased lipid peroxide levels and cholesterol oxidation products (ChOx). Formation of LDL- also takes place following Cu(2+)-induced oxidation. Cu(2+)-induced oxidation caused a small fraction of the normal unoxidized LDL (n-LDL) to convert to LDL-during the oxidative lag phase while minimal increases in conjugated dienes were apparent. After the lag phase, there was a further increase in LDL-, a rapid accumulation of conjugated dienes, and another more electronegative particle was formed (LDL2-). By the end of the lag phase, approximately 30% and 12% of the total LDL converted to LDL- and LDL2-, respectively. Nearly 40% of the total ChOx formed was present by the end of the lag period, accompanied by small increases in conjugated dienes. The major products accumulating during this time were 7-ketocholesterol, cholesterol-beta epoxide and 7-alpha-hydroxycholesterol. Accumulation of predominated during the subsequent propagation phase. At the end of propagation phase there was a six fold increase in conjugated dienes and total ChOx increased eight-fold. It appears that a subpopulation of LDL rapidly converts to LDL-, representing a mildly oxidized but oxidant sensitive LDL population. Oxidation of cholesterol accompanies these early events in LDL oxidation with formation of specific ChOx. PMID- 9199884 TI - Uric acid and ascorbic acid redox ratios in plasma and tracheal aspirate of preterm babies with acute and chronic lung disease. AB - This study compared plasma redox ratios of uric acid and ascorbic acid in well preterm babies with those with respiratory distress syndrome (RDS) and chronic lung disease (CLD), and investigated the relationship between these ratios and their respective measurements in tracheal aspirate. On day 1 after birth, plasma allantoin and allantoin/uric acid ratio were elevated in CLD (p < .05), and both markers of oxidative stress enabled early prediction of development of CLD (sensitivity and specificity: 54 and 83%, respectively). The relation between allantoin production and oxidative stress is supported by the correlation between the allantoin level and oxygen therapy in both RDS and CLD (p < .05). Reduced and oxidize ascorbic acid in plasma decreased postnatally in all groups and their redox ratio remained stable. Uric acid and ascorbic acid redox ratios were significantly elevated in tracheal aspirates compared to plasma samples (p < .05), and there was a strong positive correlation between both ratios (p < .005). These markers may be useful in monitoring babies with respiratory distress. PMID- 9199883 TI - Inactivation of cathepsin B by oxidized LDL involves complex formation induced by binding of putative reactive sites exposed at low pH to thiols on the enzyme. AB - We recently showed that the poor degradation of apo B in oxidized (ox-) LDL by mouse peritoneal macrophages could be attributed to the inactivation of cathepsin B by ox-LDL. In this current study, we show that enzyme inactivation involves complex formation of ox-LDL with cathepsin B rather than the diffusion of reactive components from ox-LDL to the enzyme. Complex formation between ox-LDL and cathepsin B was far greater at pH 4.5 than at pH 7.4 and far greater with ox LDL than with LDL. Even though complexes were also formed between ox-LDL and other proteins such as BSA, insulin, and LDL, ox-LDL bound up to 30 times more cathepsin B than BSA, when compared on a molar level and under the same conditions. Unlike ox-LDL alone, complexes of ox-LDL and BSA were unable to inactive cathepsin B, suggesting that BSA was sequestering reactive sites on ox LDL. The interaction of ox-LDL with proteins such as cathepsin B appears to represent aldehydic modifications of apo B, since treatment of ox-LDL with the reductant NaBH4, which stabilizes such adducts, greatly decreased the binding of ox-LDL to BSA and prevented ox-LDL from inactivating cathepsin B. It is likely that thiols on cathepsin B or other proteins interact with reactive groups on ox LDL, since BSA in which thiols were blocked with N-ethylmaleimide (NEM), failed to bind to ox-LDL. Moreover, NEM-treated BSA had no effect on the ability of ox LDL to inactivate cathepsin B. Similar results were obtained with LDL modified with 4-hydroxynonenal (HNE). These data suggest that aldehydic adducts on ox-LDL that are unreactive at neutral pH, possibly HNE bound to apo B, become exposed at acidic pH and then covalently bind thiols on neighboring proteins such as cathepsin B in lysosomes, inducing crosslinking of proteins and enzyme inactivation. PMID- 9199886 TI - Role of lipid peroxidation and antioxidants in gastric mucosal injury induced by the hypoxanthine-xanthine oxidase system in rats. AB - Free radical-induced gastric mucosal injury was caused by severe depletion of glutathione and alpha-tocopherol. Intravenous infusion of hypoxanthine (HX) via the jugular vein and local intra-arterial infusion of xanthine oxidase (XO) via the left gastric artery caused marked gastric mucosal injury in the antrum and the corpus. This study was performed to determine whether antioxidants in the gastric mucosa are mobilized during oxidative stress in the rat stomach. The level of thiobarbituric acid (TBA) reactive substance in the gastric mucosa was not significantly changed. The levels of total glutathione and alpha-tocopherol in the gastric mucosa significantly decreased. Total superoxide dismutase (Cu/Zn and Mn-SOD) and glutathione peroxidase activities were not significantly changed. Administration of SOD reversed the glutathione level but not the alpha-tocopherol level in the gastric mucosa. To determine the role of glutathione and alpha tocopherol in oxidative stress, the stomach was removed from a normal, alpha tocopherol supplemented, and glutathione-depleted rat and used for experimentation. Frozen slices of the rat stomach were infused with HX-XO then examined histochemically using cold Schiff's reagent for signs of lipid peroxidation. It was found that the alpha-tocopherol supplemented stomach inhibited lipid peroxidation induced by HX-XO. Biochemical measurements and histochemical examination showed that the glutathione-depleted frozen tissue section and the homogenate had increased by lipid peroxidation induced by HX-XO. These findings suggested that alpha-tocopherol and glutathione may play a role in protecting the gastric mucosa against oxygen free radicals. PMID- 9199885 TI - Green tea polyphenols inhibit oxidant-induced DNA strand breakage in cultured lung cells. AB - The influence of green tea polyphenols (GTP) on the formation of DNA strand breaks (DNA-SB) and lipid peroxidation products (LPP) in cultured human lung cells (A 549) exposed to different oxidants was investigated. Cells were pretreated with GTP for 2 h and then exposed to cigarette smoke solution, H2O2 or FeCl3 for 30 min. After exposure, the cells were analyzed for DNA-SB, LPP, and viability. In addition, the effects of GTP added directly to the incubation mixtures during exposure were examined, using the same end points. It appeared that pretreatment with GTP inhibited both cigarette smoke- and H2O2-induced DNA breakage; i.e., following exposure to cigarette smoke or H2O2, the fraction of DNA passing through a microfilter increased significantly in cells not subjected to GTP, but this effect was prevented or inhibited in GTP-treated cells. Pretreatment with GTP also reduced the overall toxicity of H2O2 as determined by cell growth after exposure. Moreover, addition of GTP during exposure reduced both cigarette smoke- and H2O2-induced DNA breakage as well as formation of LPP after exposure to Fe3+. These results indicate that GTP inhibit the formation of DNA-SB in cells exposed to oxidants. It is possible that this ability to GTP to inhibit DNA-SB formation might contribute to the antitumorogenic properties of green tea. PMID- 9199888 TI - Effect on an iron-chelator on ascorbate-induced cytotoxicity. AB - We investigated the effect of deferoxamine mesylate (DFO), an iron chelator, to test whether ascorbate-induced cytotoxicity is due to iron-catalyzed oxidation. Exposing human promyelocytic leukemic HL-60 cells to either sodium ascorbate or ascorbic acid for 1 h resulted in the progressive production of apoptotic cells characterized by cell shrinkage, as well as nuclear and internucleosomal DNA fragmentation. The addition of micromolar to millimolar concentrations of DFO during the 1-h exposure did not inhibit, but rather enhanced the ascorbate induced apoptosis in both regular and serum-free RPMI1640 medium. However, a higher concentration of serum significantly inhibited the ascorbate-induced cytotoxicity. In contrast, the cytotoxic activity of ascorbate against T98G human glioblastoma cells was enhanced or reduced by micromolar and millimolar concentrations of DFO, respectively. Ascorbate significantly increased the oxidation potential in the culture medium, and the pro-oxidant action of ascorbate was further augmented by the presence of the cells. DFO did not significantly affect the ascorbyl radical intensity and only slightly reduced the ascorbate-elevated oxidation potential. These data demonstrated that ascorbate can induce cytotoxicity even in iron-deficient medium. PMID- 9199887 TI - Generation of a polyclonal antibody against lipid peroxide-modified proteins. AB - A specific polyclonal antibody against the lipid peroxide (LOOH)-modified rabbit serum albumin (RSA) was generated in rabbits. The antibody selectively recognized the modified protein in a concentration-dependent manner and did not cross react with aldehyde-modified proteins or proteins directly oxidized with the free radical generator 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH). Oxidized low-density lipoprotein (Ox-LDL), but not native LDL, was also recognized by the antibody in a concentration-dependent manner. The antibody also cross reacted with several other proteins modified by LOOH suggesting that the antibody is directed towards a common epitope and not towards the protein sequence. Western blot analysis of normal human plasma showed that at least three different proteins are recognized by the antibody. RAW cells, preincubated with LOOH, were immunostained with the antibody and the antigenic epitopes were present intracellularly, while controls lacking in the primary antibodies failed to show immunoreactivity. Atherosclerotic arteries from cholesterol-fed monkeys and human atherosclerotic lesions were also immunostained by the antibody. The immunoreactivity was co-localized in areas rich in foam cell macrophages. These results suggest that LOOH-modified proteins present an unique antigenic epitope that may represent a primary product of interaction of LOOH with proteins. PMID- 9199889 TI - Phospholipase D activity in the intestinal mitochondria: activation by oxygen free radicals. AB - A prominent feature of cell damage caused by oxidative stress is morphological and functional changes in the mitochondria. The present study looked at the effect of free radical exposure on intestinal mitochondrial lipids. Free radical exposure did not alter neutral lipids, but among the phospholipids, phosphatidylethanolamine (PE) content was decreased on exposure to superoxide anion, generated by xanthine-xanthine oxidase or menadione with a concomitant increase in the level of phosphatidic acid (PA), suggesting activation of phospholipase D (PLD). This enzyme did not show transphosphatidylation activity in the presence of ethanol or butanol, and the product formed was phosphatidic acid (PA). This was confirmed by separation of reaction products by HPLC. This alteration in mitochondrial phospholipid was abolished by the presence of superoxide dismutase. Exposure to H2O2 did not have any significant effect. Activation of PLD by free radicals was further confirmed by quantitation of ethanolamine released from PE. Absence of any change in the content of lysophospholipid or diglyceride following exposure of mitochondria to superoxide ruled out the involvement of phospholipase A2 or C in the altered lipid composition. Moreover, inclusion of phospholipase A2 inhibitors, chlorpromazine, or p-bromophenacyl bromide did not prevent the generation of PA on exposure to free radicals. These findings suggest that superoxide anion stimulates intestinal mitochondrial PLD resulting in PE degradation and PA formation. These alterations in mitochondrial lipids may play a role in causing the functional alteration seen in oxidative stress. PMID- 9199890 TI - Effect of aromatic nitroxides on hemolysis of human erythrocytes entrapped with isolated hemoglobin chains. AB - An in vitro model of thalassemia was produced by entrapment of isolated hemoglobin chains in human erythrocytes, thus subjecting the loaded cells to oxidative stress. The presence of these unpaired chains induced physico-chemical modifications at the membrane level as studied by laurdan fluorescence. The polarity of the lipid bilayer was shown to decrease with a concomitant shift towards a gel phase in alpha-loaded erythrocytes. The determination of conjugated dienes before the hemolytic event was used as an oxidation index; the results obtained demonstrate that beta thalassemia is associated with oxidative stress. Furthermore, the presence of indolinic and quinolinic nitroxide radicals, a new class of antioxidants, in suspensions of alpha-loaded erythrocytes protected the erythrocytes from the hemolytic event. However, the protective effect exerted by the nitroxide radicals is not related to effects on membrane polarity and lipid peroxidation, even though a chemiluminescence study has demonstrated the superoxide scavenging activity of these nitroxide radicals. PMID- 9199891 TI - Role of free radicals in the mechanism of the hydrazine-induced formation of megamitochondria. AB - The effect of 4-hydroxy-2,2,6,6-tetramethyl-piperidine-1-oxyl(4-OH-TEMPO), a scavenger for free radicals, and 4-hydroxypyrazolo [3,4 d(pyrimidine)allopurinol], a xanthine oxidase inhibitor, on the hydrazine-induced changes of mitochondrial ultrastructure and those in the antioxidant system of the liver were investigated using rats as experimental animals. Animals were placed on a powdered diet containing 0.5% hydrazine for 7 d in the presence and absence of a combined treatment with 4-OH-TEMPO or allopurinol. Results obtained were as follows. 4-OH-TEMPO completely prevented the hydrazine-induced formation of megamitochondria in the liver, while it was partly prevented by allopurinol. The following changes observed in hydrazine-treated animals were improved almost completely by 4-OH-TEMPO:decreases in the body weight and liver weight; lowered rates of ADP-stimulated respiration and coupling efficiency of hepatic mitochondria; remarkable elevation of the level of lipid peroxidation. Improving effects of allopurinol were incomplete. The present results suggest that free radicals may play a key role in the mechanism of the hydrazine-induced formation of megamitochondria and that a part of free radicals generated during the hydrazine intoxication is ascribed to the degradation of purine nucleotides via xanthine oxidase. A general mechanism of the megamitochondria formation induced in various pathological conditions besides the case of hydrazine are discussed. PMID- 9199892 TI - Do reactive oxygen species underlie the mechanism of apoptosis in the tadpole tail? AB - Although "programmed cell death" has been postulated to underlie the mechanism for the disappearance of the tadpole tail, critical factors that trigger this phenomenon remain to be elucidated. To investigate the mechanism for the disappearance of the tail, changes in morphology, DNA status and the enzymes which metabolize reactive oxygen species were determined in the tail of Rana rugosa tadpoles. Histological examination revealed that apoptotic cell death was apparent in the tail myocytes during metamorphosis. Electrophoretic analysis of the tail DNA revealed a marked fragmentation. During the apoptotic changes, the activity of Cu/Zn-type superoxide dismutase (SOD) in the tail markedly increased with a concomitant decrease in its catalase activity. The apoptotic process was markedly enhanced by adding hydrogen peroxide (H2O2) and aminotriazole, a catalase inhibitor, to the medium. These observations suggested that apoptotic cell death in the tadpole tail might be triggered, at least in part, by a mechanism depending on active oxygen species, such as H2O2. PMID- 9199894 TI - Transcriptional inhibition of manganese superoxide dismutase (SOD2) gene expression by DNA methylation of the 5' CpG island. AB - Manganese superoxide dismutase (MnSOD) enzyme activity and SOD2 gene expression have often been reported to decrease during the development of cancer. SOD2 has also been implicated as a candidate tumor suppressor gene for human malignant melanoma. Genomic DNA methylation patterns are also known to change during carcinogenesis and serve as a mechanism for tumor suppressor gene inactivation. We hypothesized that decreased SOD2 gene expression in some malignant cell populations may be due, at least in part, to methylation of upstream transcriptional regulatory sequences in the SOD2 gene. To test this hypothesis we transfected methylated and unmethylated SOD/2-CAT promoter-reporter constructs in cells known to express the SOD2 gene. Our results indicate that methylation of specific cytokines in the SOD2 5' flanking region is sufficient to repress transcriptional activity of the SOD2 promoter by at least 50%. Moreover, we show that this transcriptional repression was likely mediated by inhibition of AP-2 DNA binding and transactivation from a methylated AP-2 binding site in the SOD2 promoter. DNA methylation may provide a mechanism for transcriptional inactivation of the SOD2 gene during the development of some cancers. PMID- 9199893 TI - Antioxidant constituents from licorice roots: isolation, structure elucidation and antioxidative capacity toward LDL oxidation. AB - The present study analyzed the antioxidative properties of natural compounds from the root of the plant Glycyrrhiza glabra (licorice) toward LDL oxidation. Seven constituents, with antioxidant capacity were isolated from Glycyrrhiza glabra. The isolated compounds were identified as the isoflavans Hispaglabridin A (1), Hispaglabridin B (4), Glabridin (3), and 4'-O-Methylglabridin (2), the two chalcones, isoprenylchalcone derivative (5) and Isoliquiritigenin (6), and the isoflavone, Formononetin (7). Among these compounds, Glabridin constituted the major amount in the crude extract (11.6%, w/w) as detected by high-performance liquid chromatography (HPLC) analysis. The antioxidative capacities of the isolated compounds (1-7) were tested against beta-carotene destruction and LDL oxidation. The isoflavans (1-4) at a concentration of 50 microM inhibited beta carotene consumption, following 90 min of incubation at 50 degrees C, similar to the inhibitory effect of the whole licorice crude extract (at 16 mg/1). The chalcones (5 and 6) exhibited moderate inhibition and the isoflavone 7 was almost inactive, whereas vitamin E (50 microM) completely inhibited beta-carotene consumption. The inhibitory effect of the constituents 1-7, at a concentration of 30 microM on 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH)-induced LDL oxidation was determined by measuring the amount of the thiobarbituric acid reactive substances (TBARS) and the amount of lipid peroxides. While compounds 1 6 exhibited high inhibitory activity, compound 7 and vitamin E were not active. A dose-dependent inhibitory effect of Glabridin, on the formation of cholesteryl linoleate hydroperoxide (CLOOH), in an AAPH-induced LDL oxidation system was also shown. Glabridin, at 5 or 40-60 microM concentration, inhibited the CLOOH formation by 62% and 90%, respectively. These results suggest that constituents 1 6 are very potent antioxidants toward LDL oxidation with Glabridin being the most abundant and potent antioxidant. As LDL oxidation is a key event in the formation of the early atherosclerotic lesion, the use of these natural antioxidants may be proven beneficial to attenuate atherosclerosis. PMID- 9199895 TI - Effects of reactive oxygen species on the biosynthesis of 12 (S) hydroxyeicosatetraenoic acid in mouse epidermal homogenate. AB - Arachidonic acid is converted to 12-hydroxyeicosatetraenoic acid (12-HETE) in a homogenate of mouse epidermal cells. When the epidermal homogenate was preincubated with scavengers of reactive oxygen species (ROS), catalase or superoxide dismutase, significantly larger amounts of 12-HETE were produced as compared to untreated controls, suggesting that 12-lipoxygenase is quite prone to inactivation by ROS and peroxides. Mouse epidermal homogenate was then exposed to nine different ROS-generating systems to study the effects of superoxide, hydrogen peroxide, singlet oxygen, hypochlorite, peroxyl radicals, and alkyl hydroperoxides on the enzyme activity. Analysis by chiral phase high performance liquid chromatography demonstrated that the 12-HETE biosynthesized from arachidonic acid by mouse epidermal homogenate was the 12 (S)-enantiomer and excludes oxidation of arachidonic acid by ROS in a nonspecific free radical mechanism which leads to racemic 12-HETE. ROS generated by the interaction of xanthine with xanthine oxidase strongly inhibited epidermal 12 (S)-HETE biosynthesis. A flux of 0.7 nmol of superoxide/min/ml of reaction medium resulted in more than 50% inhibition of epidermal 12-lipoxygenase activity. The decrease in 12 (S)-HETE biosynthesis appeared to involve both superoxide and hydrogen peroxide. The efficacy of the latter species was also documented by exposure of mouse epidermal 12-lipoxygenase to glucose and glucose oxidase, which resulted in similar inhibitory effects on 12 (S)-HETE biosynthesis. The presence of the iron chelator diethylenetriaminepentaacetic acid during incubation of epidermal 12 lipoxygenase with both the xanthine/xanthine oxidase or the glucose/glucose oxidase systems partially protected the enzyme against inhibition, indicating that hydroxyl radical contributes to the overall inhibitory effect. Also, organic hydroperoxides inhibited epidermal 12-lipoxygenase, whereas singlet oxygen, hypochlorite, and peroxyl radicals were not effective. The results of this study lead to the proposal that 12-lipoxygenase activity may be regulated by ROS such as hydrogen peroxides, superoxide, and hydroxyl radicals. PMID- 9199896 TI - The catalytic role of carbon dioxide in the decomposition of peroxynitrite. AB - The fast reaction of peroxynitrite with CO2 and the high concentration of dissolved CO2 in vivo (ca. 1 mM) suggest that CO2 modulates most of the reactions of peroxynitrite in biological systems. The addition of peroxynitrite to CO2 produces of the adduct ONOO-CO2- (1). The production of 1 greatly accelerates the decomposition of peroxynitrite to give nitrate. We now show that the formation of 1 is followed by reformation of CO2 (rather than another carbonate species such as CO3 = or HCO3-). To show this, it is necessary to study systems with limiting concentrations of CO2. (When CO2 is present in excess, its concentration remains nearly constant during the decomposition of peroxynitrite, and the recycling of CO2, although it occurs, can not be detected kinetically). We find that CO2 is a true catalyst of the decomposition of peroxynitrite, and this fundamental insight into its action must be rationalized by any in vivo or in vitro reaction mechanism that is proposed. When the concentration of CO2 is lower than that of peroxynitrite, the reformation of CO2 amplifies the fraction of peroxynitrite that reacts with CO2. Even low concentrations of CO2 that result from the dissolution of ambient CO2 can have pronounced catalytic effects. These effects can cause deviations from predicted kinetic behavior in studies of peroxynitrite in noncarbonate buffers in vitro, and since 1 and other intermediates derived from it are oxidants and/or nitrating agents, some of the reactions attributed to peroxynitrite may depend on the availability of CO2. PMID- 9199897 TI - Proteins but not nucleic acids are molecular targets for the free radical attack during reoxygenation of rat hepatocytes. AB - Isolated rat hepatocytes generate large amounts of reactive oxygen species and suffer a significant cell injury during postanoxic reoxygenation. The aim of this study was to determine whether oxidation of proteins and nucleic acids occurs during reoxygenation and whether their damage is related to the development of hepatocyte injury. Isolated perfused rat hepatocytes were exposed sequentially to 1 h of aerobic control, 2.5 h of anoxia, and 2 h of reoxygenation. Protein oxidation was determined by measuring the hepatocyte protein carbonyl content. DNA and RNA oxidation was assessed by measuring the 8-hydroxydeoxyguanosine and 8 hydroxyguanosine adducts, respectively. The control preanoxic carbonyl content was 6.48 +/- 1.03 nmol/mg protein. The preanoxic 8-8 hydroxydeoxyguanosine and 8 hydroxyguanosine levels were 4.76 +/- 1.22 pmol/ml and 14.19 +/- 2.17 pmol/ml, respectively. During anoxia, protein and nucleic acid oxidation did not change significantly. With reoxygenation, the protein carbonyl content increased significantly within 30 min, reaching a value of 10.25 +/- 1.58 nmol/mg. The nucleic acid oxidation level remained stable. Perfusion with 100 muM of during reoxygenation abolished protein oxidation. These results indicate that in rat hepatocytes during the early phase of reoxygenation: (1) the protein oxidation level increased significantly above the preanoxic aerobic values; (2) DNA and RNA oxidation does not appear to occur; and (3) free metal-mediated free radical reactions are involved in the oxidative protein damage. PMID- 9199898 TI - S-allyl cysteine inhibits activation of nuclear factor kappa B in human T cells. AB - Reactive oxygen species are involved in signal transduction pathways leading to nuclear factor kappa B (NF-kappa B) activation which has been implicated in the regulation of gene transcription. We recently reported that a garlic compound, S allyl cysteine (SAC), protects bovine pulmonary artery endothelial cells from oxidant injury induced by hydrogen peroxide (H2O2). In this study we determined the effects of SAC on NF-kappa B activation in human T lymphocytes (Jurkat cells) induced by tumor necrosis factor alpha (TNF- alpha) and H2O2. Activated NF-kappa B in nuclear extracts was measured by an electrophoretic mobility shift assay using 32P-labeled probe. SAC consistently exhibited a dose-dependent inhibition of NF-kappa B activation induced by both TNF-alpha and H2O2. Supershift with specific antibodies to NF-kappa B subunits confirmed that the inducible retarded bands observed in the EMSA and p65-p50 heterodimer of the NF-kappa B/Rel protein. Our data suggest that SAC may act via antioxidant mechanisms to block NF-kappa B activation in Jurkat cells. PMID- 9199899 TI - Failure of colonoscopy to detect colorectal cancer: evaluation of 47 cases in 20 hospitals. AB - BACKGROUND: Colonoscopy is the gold standard for the detection of colon polyps and cancers, but failed detections can occur and the reasons are incompletely understood. METHODS: During a retrospective evaluation of the sensitivity of barium enema and colonoscopy in 20 Indiana Hospitals, we encountered 47 cases of colorectal cancer in which a colonoscopy performed within 3 years of the diagnosis had not detected the cancer. Cases were reviewed for location of tumor, extenuating factors, pathologic features, delay in diagnosis from failed detection, and who performed the examination. RESULTS: Failed detection was more likely when colonoscopy was performed by a nongastroenterologist than a gastroenterologist (odds ratio 5:36, 95% CI [2.94,9.77]). Twenty-seven cancers were "missed," and 20 were estimated to be not reached. However, the location of missed tumors and a general absence of adequate documentation of cecal intubation suggested that some cecal and ascending colon cancers recorded as missed may actually have been not reached. Variation in sensitivity among gastroenterologists suggested that meticulous examination is also important in maximizing sensitivity. CONCLUSIONS: These cases suggest several factors that might improve the quality and sensitivity of colonoscopy: (1) examiners should receive adequate training, (2) cecal intubation rates should be high, (3) cecal intubation should be verified by specific landmarks in all cases, (4) failure to reach the cecum should be followed by prompt barium enema, and (5) meticulous examination would appear to improve sensitivity for cancer detection. PMID- 9199900 TI - Extent of Barrett's metaplasia: a prospective study of the serial change in area of Barrett's measured by quantitative endoscopic imaging. AB - BACKGROUND: An accurate determination of the extent of Barrett's metaplasia is critical to the study of its natural history and response to therapy. Our hypothesis is that area calculations offer advantages over length estimates of Barrett's. METHODS: Changes in both measures and estimates of progression or regression between two endoscopies in 17 patients were compared. Area was calculated using a computer image analysis technique. RESULTS: Although there was no significant difference in length correlation versus area correlation between endoscopies (r = 0.90 vs 0.99), the mean change in absolute length (1.4 +/- 0.2 cm) was greater than the change in area (4.5 +/- 1.4 cm2, equivalent to a length of 0.67 +/- 0.2 cm, p = 0.001). The percent change in absolute length (26.9%) was greater than the change in area (16%, p = 0.001). Discordance of estimates of progression or regression between area and length was found in nine patients. The image technique detected no change in the area of squamous islands. CONCLUSIONS: Imaging analysis can precisely measure the extent of Barrett's including squamous islands. Area showed little change, whereas measures of length were more varied. Computer based image analysis provides a more precise estimate of interval change of Barrett's. PMID- 9199901 TI - Prospective comparison of H&E, Giemsa, and Genta stains for the diagnosis of Helicobacter pylori. AB - BACKGROUND: H. pylori is more easily visualized with special stains than with H&E, but this adds time and expense to the diagnostic workup. We sought to determine if the diagnostic accuracy was improved with special stains. METHODS: One hundred-one patients had two "jumbo" biopsies taken from the gastric antrum and two from the body for examination with H&E, Genta, and Giemsa stains. Four separate biopsy specimens were also taken from the antrum and the body for culture and for three types of rapid urease test, and 13C-urea breath tests were also performed. Mixed, coded biopsies were assessed for H. pylori, and density was scored from 0 to 4. A case was considered positive for H. pylori if culture was positive, two rapid urease tests and a urea breath test were positive, or two different stains were positive. Biopsy specimens were excluded from analysis if the slides were missing or there was inadequate tissue for review, or if the specimen showed a lack of staining. RESULTS: Fifty-two (13%) of 404 specimens were excluded because of a poor Genta stain. Sensitivities were comparable for the three stains (H&E, 92%; Giemsa, 88%; Genta, 91%), while H&E specificity (89%) was significantly lower than that of the special stains (98%). Sensitivity for all three stains was significantly lower at low (grade 0 to 1) H. pylori density than at high (grade 2 to 4) density (H&E, 70% vs 98%; Giemsa, 64% vs 96%; Genta, 66% vs 97%), and 20 of 22 false positives were grade 1. CONCLUSIONS: The sensitivities of H&E and special stains are comparable at around 90%, but the specificity of H&E is significantly lower. The Giemsa stain appears to be the preferred stain for H. pylori diagnosis on the basis of its good sensitivity, excellent specificity, and lack of technical difficulty in preparation. However, H&E provides excellent accuracy when more than minimal (grade 1) H. pylori density is present. PMID- 9199902 TI - Endosonographic differentiation of benign and malignant stromal cell tumors. AB - BACKGROUND: Endosonography (EUS) is a valuable technique for diagnosing gastrointestinal stromal cell tumors. However, EUS features that are predictive of malignancy in these tumors have not been defined. METHODS: Videotapes and photographs of EUS examinations performed prior to surgical resection of 35 stromal cell tumors (9 malignant) were blindly reviewed by a single examiner. EUS features associated with malignancy were determined. Interobserver agreement in interpreting these features was then measured among a panel of five expert endosonographers who judged EUS videotapes of 35 resected stromal cell tumors (10 malignant). RESULTS: Stepwise logistic regression analysis demonstrated that tumor size (diameter > 4 cm), irregular extraluminal border, echogenic foci, and cystic spaces were independently associated with malignancy in stromal cell tumors (p < 0.05). Interobserver agreement for irregular extraluminal border, echogenic foci, and cystic spaces, as measured by mean kappa statistic, was 0.43, 0.39, and 0.28, respectively. For the five experts, the sensitivity for detecting malignancy ranged between 80% to 100% when at least two of the three features were judged to be present. The likelihood of finding malignancy ranged between 0% to 11% for the experts when all three features were judged absent. CONCLUSIONS: Tumor size and certain EUS features are useful for predicting malignancy in stromal cell tumors. Absence of these features indicates benign disease. Agreement among experts in interpreting these EUS features is fair to moderate. PMID- 9199903 TI - A comparison of the accuracy of echo features during endoscopic ultrasound (EUS) and EUS-guided fine-needle aspiration for diagnosis of malignant lymph node invasion. AB - BACKGROUND: The purpose of this study was to re-evaluate echo features of lymph nodes during endoscopic ultrasound and assess the utility of these echo features and endoscopic ultrasound-guided fine-needle aspiration in predicting malignant lymph node invasion. METHODS: Thirty-five lymph nodes in 25 patients with lung, esophageal, and pancreatic cancer were evaluated by endoscopic ultrasound. Endoscopic ultrasound examinations were performed with a radial scanning echoendoscope. Confirmation of benign lymph nodes was obtained by surgical resection while malignant lymph nodes were confirmed by real-time endoscopic ultrasound-guided fine-needle aspiration with a linear array echoendoscope. RESULTS: Nineteen benign lymph nodes and 16 malignant lymph nodes in the mediastinum, celiac axis, and the peripancreatic area were included in the study. The following echo features were compared between benign and malignant lymph nodes: size greater than 1 cm, hypoechoic, distinct margins, and round shape. No single feature independently predicted malignant invasion. When all four of the above features were present in the same lymph node, the accuracy for predicting malignant invasion was 80%. However, all four features of malignant involvement were present in only 25% (4 of 16) of malignant lymph nodes. Our study also suggests that the above echo features may be a less reliable predictor of malignant invasion in pulmonary malignancies when compared to gastrointestinal cancers. Endoscopic ultrasound-guided fine-needle aspiration of lymph nodes in 22 patients revealed malignant lymph node invasion in 16 and benign cells in 6 patients. CONCLUSION: Endoscopic ultrasound-guided fine-needle aspiration is an important adjunct for accurate lymph node assessment for malignancy. PMID- 9199904 TI - Endoscopic detection of early esophageal cancer in a high-risk population: does Lugol staining improve videoendoscopy? AB - BACKGROUND: The aim of this study was to prospectively compare the diagnostic accuracy of videoendoscopy, with and without Lugol staining, for the detection of esophageal cancer in alcoholic or smoking patients older than 40 years. METHODS: Daily alcohol and tobacco consumption and overt and latent symptoms were noted. The 158 patients included were examined by videoendoscopy and with Lugol dye. RESULTS: The mean consumption of alcohol and tobacco was 86 +/- 49 gm/day for 26 +/- 11 years, and 30 +/- 18 pack-years, respectively. Twenty-five patients had no symptoms. Before Lugol staining, 12 patients had endoscopically identified cancerous lesions. After Lugol staining, 13 patients had 17 esophageal cancers, 3 of which were high-grade dysplasia. The prevalence of esophageal cancer was 8.2%: 95% CI [4,14]. Dye-free surfaces were significantly larger than the endoscopic patterns observed before Lugol staining (11.6 +/- 9.2 cm2 vs 1.4 +/- 1.7 cm2; p < 0.02). CONCLUSIONS: In an alcoholic smoking population, the prevalence of esophageal cancer detected by endoscopy is high and not related to symptoms described by patients. Lugol staining only moderately improves the diagnostic accuracy of videoendoscopy; its main advantage is the assessment of the mucosal extension of esophageal cancer. PMID- 9199905 TI - Vertical lines in distal esophageal mucosa (VLEM): a true endoscopic manifestation of esophagitis in children? AB - BACKGROUND: We observed an endoscopic abnormally in a group of children with histological esophagitis. We termed this finding "vertical lines in esophageal mucosa" (VLEM). We examined the relationship between the presence of VLEM and significant histologic changes in esophageal mucosal biopsies. METHODS: Between January 1, 1992, and August 31, 1994, the senior author (JFF) performed 255 esophageal biopsies. The procedure reports, available endoscopic photographs, and histology reports were reviewed to establish the endoscopic and histologic appearance of the esophageal mucosa. Intraepithelial cells were counted in a blind review of 42 randomly selected biopsies. RESULTS: The esophageal mucosa had a normal appearance on 160 endoscopic studies (Group 1) and VLEM were the only mucosal abnormalities in 41 endoscopies (Group 2). Histology was normal in 92 of 160 biopsies (57.5%) from Group 1, and 1 of 41 biopsies (2.4%) from Group 2. Most patients in Group 2 had eosinophilic esophagitis (34 of 41, 83%, specificity 0.85, sensitivity 0.5, p > 0.001) which was of moderate to severe intensity (31 of 34, 91.2%, specificity 0.88, sensitivity 0.73, p < 0.001). CONCLUSIONS: Histology usually demonstrated moderate to severe inflammation when VLEM were present. VLEM may be a highly specific endoscopic feature of esophagitis in children. PMID- 9199906 TI - Device choice and experience level in endoscopic foreign object retrieval: an in vivo study. AB - BACKGROUND: Successful foreign object retrieval may depend on device choice and the experience level of the endoscopist, although these factors have not been systematically evaluated. METHODS: In anesthetized pigs, the ability to retrieve foreign objects (metal tack, button disc battery, wooden toothpick) placed endoscopically into the stomach was assessed. Seven university medical center gastroenterology attending physicians (5 clinical and 2 basic science research [BSR]), and 4 fellows-in-training participated. The devices used were the Roth retrieval net, rat tooth forceps, Dormia basket, polypectomy snare, and radial jaw forceps. The time to retrieve each object into an esophageal overtube within a 5 minute maximum was measured. RESULTS: Only the Roth net and Dormia basket were successful in retrieving the button disc battery, although the Roth net was superior (100% vs 27%, Fisher p < 0.025). All devices were equally successful at retrieving the tack (82% to 100%, p = NS). The snare was significantly faster than the Roth net (p < 0.05). For the tack, there was significantly fewer difficulties encountered with the snare than the Roth net (Fisher p < 0.03). The Roth net was incapable of retrieving the toothpick; the other devices were equally successful (91% to 100%). The clinical attendings had a significantly higher success rate (95%) than the fellows (82%, chi squared p < 0.05) or combined fellows/BSR attendings (80%, p < 0.02), and were significantly faster than the fellows (p < 0.0002) or the fellows/BSR attendings (p < 0.0003). CONCLUSIONS: The Roth net is the best device for retrieving smooth objects such as the button disc battery. For sharp objects, such as the tack and toothpick, best results were achieved with the snare, although the forceps were also effective. More experienced endoscopists had higher success rates and faster retrieval times. Both device choice and the experience level of the endoscopists have an impact on successful foreign object retrieval. PMID- 9199907 TI - Current induction in a fiberglass guidewire compared to conventional wires during simulated papillotomy. AB - BACKGROUND: Current induced in a guidewire during papillotomy poses a danger of injury to the bile duct. We measured currents induced in three commercially available guidewires and a prototype fiberglass wire during simulated sphincterotomy under standard and nonstandard conditions. METHODS: Blended current at 55 W was applied to a double-lumen papillotome grounded through a 1000 omega resistor. For extreme conditions, power was increased to 70 W using a single-lumen papillotome. Fault conditions were created with a break in the insulation at the distal end of each wire. Guidewire-induced current was measured, and safety calculations performed for adherence to accepted standards for electrosurgical devices. RESULTS: Induced current was within safety limits for all wires tested under standard conditions. With insulation faults, one of the commercially available wires was unsafe. Under extreme conditions, with or without faults, the three commercial wires produced currents ranging from 9% to 225% above acceptable levels, while only the prototype wire remained safe. CONCLUSIONS: Most guidewires contain metal cores that function as capacitors. Because its core is primarily fiberglass, the prototype wire generates less induced current under nonstandard conditions, thus achieving a greater margin of safety during wire-guided sphincterotomy. PMID- 9199908 TI - Efficacy of prophylactic sclerotherapy in patients with hepatocellular carcinoma and varices negative for the red color sign. AB - BACKGROUND: A prospective randomized controlled study was performed to evaluate the usefulness of prophylactic endoscopic sclerotherapy in patients with hepatocellular carcinoma complicated by esophageal varices. METHODS: The subjects included 58 patients with esophageal varices negative for the red color sign and hepatocellular carcinoma without tumor emboli in the portal trunk or primary portal branches. Patients were randomly assigned to prophylactic sclerotherapy (n = 29) or control (n = 29) groups, and their bleeding and survival rates were compared. RESULTS: A mean of 3.0 sclerotherapy sessions was required for complete disappearance of varices in patients receiving prophylactic sclerotherapy. During the observation period, transcatheter arterial embolization for hepatocellular carcinoma was performed more often in patients with prophylactic sclerotherapy (mean 3.8 times) than in control patients (mean 2.0 times) (p < .05). Percutaneous ethanol injection therapy was performed more often in patients with prophylactic sclerotherapy than in controls (mean 8.1 times vs 5.0 times, respectively) (p < .05). The 3-year bleeding rates were 50% for the control group and 18% for the prophylactic sclerotherapy group (p < 0.05), and the 3-year survival rates were 16% for the control group and 37% for the therapy group (p < 0.05). CONCLUSIONS: Prophylactic sclerotherapy improves survival in patients with hepatocellular carcinoma complicated by red color sign-negative esophageal varices without tumor emboli in the portal trunk or primary portal branches. PMID- 9199909 TI - Laparoscopic evaluation of liver disease in chronic renal failure prior to renal transplantation. AB - BACKGROUND: Diagnostic laparoscopy with liver biopsy has been shown to be safe and effective in the evaluation of patients with chronic liver disease. Patients with end-stage renal disease may be more prone to bleeding complications secondary to liver biopsy as a result of multiple factors directly related to their underlying renal condition. METHODS AND PATIENTS: From January 1994 to June 1996, 16 patients with end-stage renal disease and hepatic dysfunction (6 women and 10 men) underwent diagnostic laparoscopy with liver biopsy prior to renal transplantation at the University of Miami School of Medicine. Laparoscopy was performed using a 5 mm video laparoscope with a left paramedian approach. The mean patient age was 46 years. Fourteen patients had chronic hepatitis C with a reactive anti-HCV by ELISA; one patient had chronic hepatitis B with reactive HBsAg, and one patient was co-infected with both hepatitis B and C viruses. RESULTS: Two patients developed hypotension related to the procedure and one patient developed an intra-abdominal hemorrhage 5 days after laparoscopy that did not require surgical intervention. Biopsy findings were as follows: 13 patients had mild chronic hepatitis; 2 patients had chronic hepatitis with bridging fibrosis; and 1 patient was cirrhotic. Prior kidney transplantation or peritoneal dialysis did not preclude the performance of laparoscopy. CONCLUSION: Diagnostic laparoscopy can be safety performed in patients with end-stage renal disease with acceptable morbidity and mortality. PMID- 9199910 TI - Endoscopic ultrasonography for colorectal cancer using submucosal saline solution injection. PMID- 9199911 TI - Endoscopic mucosal resection of gastric tumors located in the lesser curvature of the upper third of the stomach. PMID- 9199912 TI - Video time lapse endoscopy. PMID- 9199913 TI - An esophageal foreign body impaction from a Tums E-X tablet. PMID- 9199914 TI - Electrohydraulic lithotripsy of a gallstone causing gallstone ileus. PMID- 9199915 TI - A rare anomaly of the biliary tree: the interhepatic duct. PMID- 9199916 TI - Unusual endoscopic features of gastric and duodenal cryptosporidiosis in AIDS. PMID- 9199917 TI - Gastric purpura: an unusual endoscopic feature in gastrointestinal hemorrhage. PMID- 9199918 TI - Missed cancers at colonscopy: learning the hard way. PMID- 9199919 TI - Detecting, measuring, and managing Barrett's esophagus: the complete endoscopist's dilemma. PMID- 9199920 TI - Future tense: ASGE strides toward the millennium. PMID- 9199921 TI - A case of malignant melanoma in the anorectal region: colonoscopic features. PMID- 9199922 TI - Inflation and positioning of the gastric balloon of a Sengstaken-Blakemore tube under ultrasonographic control. PMID- 9199923 TI - PEGS: buttons, balloons, and mushrooms. PMID- 9199924 TI - Management of Ogilvie's syndrome. PMID- 9199925 TI - Endoscopic injection of ethanolamine as a treatment for achalasia: a first report. PMID- 9199927 TI - Risk of colorectal adenomas in patients with a family history of colorectal cancer: some implications for screening programmes. PMID- 9199926 TI - Endoscopic stenting in obstructive jaundice due to liver metastases: does it have benefit for the patient? PMID- 9199929 TI - Single-minded--two genes, three chromosomes. PMID- 9199931 TI - Microsatellite data support an early population expansion in Africa. AB - We have developed a method for the analysis of microsatellite data that is useful in the elucidation of the demographic history of populations. This method, the PK distribution method of pairwise comparisons, is analogous to the mismatch distribution of sequence comparisons developed for the analysis of mitochondrial sequence data by Rodgers and Harpending and is defined as the distribution of the number of repeat unit differences between alleles when each allele in a sample is compared with every other allele in the sample. Using computer simulations of microsatellite loci, we show that the shape of the distribution of PK changes in a distinctive manner as a function either of time since population expansion or effective population size. Increases in both of these affect the PK distribution in a similar fashion leading to a change from a steep distribution with a Po peak to one with a nonzero peak. Analysis of three data sets from surveys of microsatellite loci in ethnographically defined populations reveals that most (9/12) of the African populations analyzed, but none of the 30 non-African populations showed PK distributions with nonzero peaks. These PK distributions indicate either an earlier expansion or a larger effective population size for African populations. This observation is consistent with the hypothesized African origin of modern human. PMID- 9199930 TI - Genomic organization of the human PEX gene mutated in X-linked dominant hypophosphatemic rickets. PMID- 9199932 TI - Ataxia-telangiectasia locus: sequence analysis of 184 kb of human genomic DNA containing the entire ATM gene. AB - Ataxia-telangiectasia (A-T) is an autosomal recessive disorder involving cerebellar degeneration, immunodeficiency, chromosomal instability, radiosensitivity, and cancer predisposition. The genomic organization of the A-T gene, designated ATM, was established recently. To date, more than 100 A-T associated mutations have been reported in the ATM gene that do not support the existence of one or several mutational hotspots. To allow genotype/phenotype correlations it will be important to find additional ATM mutations. The nature and location of the mutations will also provide insights into the molecular processes that underly the disease. To facilitate the search for ATM mutations and to establish the basis for the identification of transcriptional regulatory elements, we have sequenced and report here 184,490 bp of genomic sequence from the human 11q22-23 chromosomal region containing the entire ATM gene, spanning 146 kb, and 10 kb of the 5'-region of an adjacent gene named E14/NPAT. The latter shares a bidirectional promoter with ATM and is transcribed in the opposite direction. The entire region is transcribed to approximately 85% and translated to 5%. Genome-wide repeats were found to constitute 37.2%, with LINE (17.1%) and Alu (14.6%) being the main repetitive elements. The high representation of LINE repeats is attributable to the presence of three full-length LINE-1s, inserted in the same orientation in introns 18 and 63 as well as downstream of the ATM gene. Homology searches suggest that ATM exon 2 could have derived from a mammalian interspersed repeat (MIR). Promoter recognition algorithms identified divergent promoter elements within the CpG island, which lies between the ATM and E14/NPAT genes, and provide evidence for a putative second ATM promoter located within intron 3, immediately upstream of the first coding exon. The low G+C level (38.1%) of the ATM locus is reflected in a strongly biased codon and amino acid usage of the gene. PMID- 9199933 TI - Minisequencing: a specific tool for DNA analysis and diagnostics on oligonucleotide arrays. AB - We describe a method for multiplex detection of mutations in which the solid phase minisequencing principle is applied to an oligonucleotide array format. The mutations are detected by extending immobilized primers that anneal to their template sequences immediately adjacent to the mutant nucleotide positions with single labeled dideoxynucleoside triphosphates using a DNA polymerase. The arrays were prepared by coupling one primer per mutation to be detected on a small glass area. Genomic fragments spanning nine disease mutations, which were selected as targets for the assay, were amplified in multiplex PCR reactions and used as templates for the minisequencing reactions on the primer array. The genotypes of homozygous and heterozygous genomic DNA samples were unequivocally defined at each analyzed nucleotide position by the highly specific primer extension reaction. In a comparison to hybridization with immobilized allele-specific probes in the same assay format, the power of discrimination between homozygous and heterozygous genotypes was one order of magnitude higher using the minisequencing method. Therefore, single-nucleotide primer extension is a promising principle for future high-throughput mutation detection and genotyping using high density DNA-chip technology. PMID- 9199934 TI - Cloning of two human homologs of the Drosophila single-minded gene SIM1 on chromosome 6q and SIM2 on 21q within the Down syndrome chromosomal region. AB - As part of our effort to clone genes of human chromosome 21 that may contribute to Down syndrome, we have previously isolated four exons with homology to Drosophila single-minded (sim) gene, which encodes a transcription factor that is a master regulator of fruit fly neurogenesis. These exons were used to clone and characterize two human homologs of the Drosophila sim gene, SIM1 and SIM2, which map to chromosomes 6q16.3-q21 and 21q22.2, respectively; SIM2 maps within the so called Down syndrome chromosomal region. Recently, two mouse homologs, Sim1 and Sim2, also have been identified. There is a high level of homology among human, mouse, and Drosophila sim genes in their amino-terminal half where the conserved bHLH, PAS1, PAS2, and HST domains are present. In contrast, the carboxy-terminal parts are only homologous between SIM1 and Sim1 and SIM2 and Sim2. Two isoforms (SIM2 and SIM2s) of human SIM2 have been detected that differ in their 3' ends. Northern blot analysis revealed one mRNA SIM1 species of approximately 9.5 kb and four different mRNA SIM2 species of 2.7, 3, 4.4, and 6 kb in human fetal kidney. The function of both human SIM1 and SIM2 is unknown. However, three copies of SIM2 may contribute to some specific Down syndrome phenotypes because of (1) mapping position, (2) potential function as transcriptional repressor, (3) likely dimerization with other transcription factors, (4) the temporal and spatial expression pattern of mouse Sim2, and (5) the potentially analogous role of human SIM2 to that of Drosophila sim during neurogenesis. PMID- 9199935 TI - Genomic organization of the murine Miller-Dieker/lissencephaly region: conservation of linkage with the human region. AB - Several human syndromes are associated with haploinsufficiency of chromosomal regions secondary to microdeletions. Isolated lissencephaly sequence (ILS), a human developmental disease characterized by a smooth cerebral surface (classical lissencephaly) and microscopic evidence of incomplete neuronal migration, is often associated with small deletions or translocations at chromosome 17p13.3. Miller-Dieker syndrome (MDS) is associated with larger deletions of 17p13.3 and consists of classical lissencephaly with additional phenotypes including facial abnormalities. We have isolated the murine homologs of three genes located inside and outside the MDS region: Lis1, Mnt/Rox, and 14-3-3 epsilon. These genes are all located on mouse chromosome 11B2, as determined by metaphase FISH, and the relative order and approximate gene distance was determined by interphase FISH analysis. The transcriptional orientation and intergenic distance of Lis1 and Mnt/Rox were ascertained by fragmentation analysis of a mouse yeast artificial chromosome containing both genes. To determine the distance and orientation of 14 3-3 epsilon with respect to Lis1 and Mnt/Rox, we introduced a super-rare cutter site (VDE) that is unique in the mouse genome into 14-3-3 epsilon by gene targeting. Using the introduced VDE site, the orientation of this gene was determined by pulsed field gel electrophoresis and Southern blot analysis. Our results demonstrate that the MDS region is conserved between human and mouse. This conservation of linkage suggests that the mouse can be used to model microdeletions that occur in ILS and MDS. PMID- 9199936 TI - The complex mutation pattern of a microsatellite. AB - DQCAR is a (CA)n microsatellite located in the HLA class II region and tightly linked to HLA-DQB1. Previous studies showed a strikingly low level of size variation in DQCAR alleles within an extensive subfamily of HLA-DQ subtypes (DQ1). DQCAR alleles in non-DQ1 subtypes showed a higher degree of size polymorphism. In this study sequence analysis demonstrates that DQ1-associated DQCAR alleles have a single C-->A nucleotide substitution interrupting the CA repeat array. Frequent CA-->GA mutations are also observed in DQ1-associated microsatellites with identical allele sizes. In contrast, DQCAR alleles associated with non-DQ1 haplotypes display a perfect CA repeat sequence and the variation in allele size is attributable only to differences in the number of CA repeats. Our results imply that several mutational mechanisms are involved in the generation of allelic diversity within the same microsatellite locus. The possibility of different mutation rates in the same locus should to be taken into account when using these markers in evolutionary and disease studies. PMID- 9199937 TI - Sequencing and functional analysis of the SNRPN promoter: in vitro methylation abolishes promoter activity. AB - The gene encoding the small nuclear ribonucleoprotein-associated polypeptide N (SNRPN) maps to the Prader-Willi syndrome critical region on chromosome 15 and is expressed preferentially from the paternal allele. A CpG island encompassing the first exon of SNRPN is methylated on the inactive maternal allele. DNA sequence was determined for a cosmid containing the first three exons of SNRPN and extending 20 kb upstream and 15 kb downstream from the CpG island. This region is extremely rich in Alu elements and other repetitive sequences and contains a single CpG island, which includes numerous short direct repeat sequences. Functional analysis of the first exon revealed strong promoter activity for a 260 bp fragment extending 207 bp upstream from the exon. In vitro methylation of this 260-bp fragment abolished promoter activity completely, suggesting that the silencing of the maternal SNRPN allele may be a direct consequence of methylation of the promoter region. PMID- 9199938 TI - PowerBLAST: a new network BLAST application for interactive or automated sequence analysis and annotation. AB - As the rate of DNA sequencing increases, analysis by sequence similarity search will need to become much more efficient in terms of sensitivity, specificity, automation potential, and consistency in annotation. PowerBLAST was developed, in part, to address these problems. PowerBLAST includes a number of options for masking repetitive elements and low complexity subsequences. It also has the capacity to restrict the search to any level of NCBI's taxonomy index, thus supporting "comparative genomics" applications. Postprocessing of the BLAST output using the SIM series of algorithms produces optimal, gapped alignments, and multiple alignments when a region of the query sequence matches multiple database sequences. PowerBLAST is capable of processing sequences of any length because it divides long query sequences into overlapping fragments and then merges the results after searching. The results may be viewed graphically, as a textual representation, or as an HTML page with links to GenBank and Entrez. For matching database sequences, annotated features are superimposed on the aligned query sequence in the output, thus greatly increasing the ease of interpretation. Such features may be used for automated annotation of new sequence because PowerBLAST output in ASN.1 form may be "dragged and dropped" into NCBI's Sequin program for sequence annotation and submission. PowerBLAST is capable of analyzing and annotating a 100-kb query in 60 min on NCBI's BLAST server. PMID- 9199939 TI - Centromeric and noncentromeric ADE2-selectable fragmentation vectors for yeast artificial chromosomes in AB1380. AB - We have constructed a set of fragmentation vectors for the truncation of either the centromeric or the noncentromeric end of YACs containing a human DNA insert. These vectors carry ADE2 or HIS5 as the selectable marker, enabling direct use in AB1380, the host strain of most publicly available YAC libraries. Centromeric fragmentation vectors for AB1380 have not been reported previously; the noncentromeric vectors show high frequencies of fragmentation. PMID- 9199940 TI - Coronary stenting in 1000 consecutive patients. Long-term clinical and angiographic results. AB - Coronary stent implantation has become an accepted treatment for selected patients but the long-term outcome after stent implantation has not been investigated in a large unselected population. This study reports clinical and angiographic results after coronary stent implantation in a consecutive group of 1000 patients treated between November 1989 and June 1994. A total of 2012 stents were implanted in 1216 lesions. The anticoagulation regimen after successful stenting was abolished in the last 499 patients, treated with aspirin and ticlopidina. Complex lesions (type B2, type C lesions and chronic total occlusions) were less frequent in the group with anticoagulation (67%, 355/529 lesions) than in the group without anticoagulation (76%, 485/641; p < 0.01). Vessel size was also significantly smaller in the group with no anticoagulation (3.21 +/- 0.50 mm vs 2.89 +/- 0.70 mm; p < 0.001). Procedural success was achieved in 96% of cases. Major complications occurred in 49 pts, including death (1%), emergency bypass surgery (2.7%), and myocardial infarction (2.1%). Subacute stent thrombosis, observed in 2.6% of the patients treated with anticoagulation, decreased to 1% in the last 499 patients receiving only antiplatelet drugs after high pressure stent expansion controlled with intravascular ultrasound (p < 0.05). Angiographic follow-up was obtained at a mean interval of 6.6 +/- 3.0 months in 718 patients (75% of the eligible candidates). The global restenosis rate (greater than 50% diameter stenosis) was 22%, with no statistically significant difference in patients with and without anticoagulation after stenting. Clinical follow-up was available in 890 patients at a mean interval of 14 +/- 17 months. Late cardiac events occurred in 251 patients and included: 159 repeat coronary angioplasties for restenosis (20%), 114 coronary angioplasties for de novo lesions (10%), 33 elective by-pass surgery operations (3.7%) and 42 late deaths (4.7%). At the most recent follow-up there were 719 pts (81%) free of anginal symptoms, including patients having repeat angioplasty for restenosis or disease progression. Coronary stenting has a low incidence of procedural complications and high-pressure ultrasound guided implantation has drastically reduced the risk of subacute thrombosis. Repeat procedures due to restenosis remain the most important limitation of the technique and do not seem to be affected by evolving technique and operator experience. PMID- 9199941 TI - The relative value of exercise-electrocardiography and dipyridamole stress echocardiography for risk stratification early after uncomplicated myocardial infarction. The EPIC (Echo Persantine International Cooperative) Study Group. AB - BACKGROUND: Rational prognostic algorithm should be developed considering the logical progression of the information as it becomes available to the physician, with clinical data first, ECG data second and stress imaging data last. The aim of the present study was to assess in a clinically realistic fashion the relative prognostic value of exercise electrocardiography test (EET) and dipyridamole echocardiography test (DET) early after first acute uncomplicated myocardial infarction. METHODS AND RESULTS: Five hundred and forty-seven in-hospital patients (age = 56 +/- 9 years) with recent clinically uncomplicated first myocardial infarction, baseline echocardiographic findings of satisfactory quality, interpretable ECG and capability to exercise underwent a resting 2D echocardiogram, a DET and an EET at a mean of 10 days from the infarction and were followed up for 16.2 +/- 11 months. During the follow-up, there were 17 cardiac deaths, 19 non-fatal myocardial infarctions and 49 unstable angina. When cardiac death was considered as the only significant event, with multivariate analysis, peak dipyridamole Wall Motion Score Index was the only significant predictor (chi 2 = 5.66; p = 0.013; relative risk estimate = 4.7; confidence intervals = 1.35-16.08). In presence of a negative exercise electrocardiography test for both chest pain and electrocardiographic criteria, the death rate was 2%. CONCLUSION: DET provides stronger information in comparison with historical and EET variables. However, a negative maximal EET is sufficient to identify a very low risk subset in whom additional testing may not be warranted. PMID- 9199942 TI - Electrocardiographic Minnesota code findings predicting short-term mortality in asymptomatic subjects. The Italian RIFLE Pooling Project (Risk Factors and Life Expectancy). AB - Aim of the study was to analyze the predictive power on short term mortality of electrocardiographic findings in asymptomatic subjects belonging to samples of the general population. In the Italian RIFLE Pooling Project (Risk Factors and Life Expectancy) 12 180 men and 10 373 women aged 30 to 69 years had a resting electrocardiogram (ECG) recorded at baseline examination. All of them were free from clinically symptomatic heart disease and represented 23 cohorts spread all over Italy. ECGs were read by the Minnesota Code using 5 large categories of abnormalities, i.e. Q-QS abnormalities, ST-T abnormalities, high R. waves, major arrhythmias, and blocks. Some clinically relevant ECG combination of abnormalities were also analyzed. Six-year mortality from coronary heart disease (CHD), cardiovascular diseases (CVD) and all-cause mortality (ALL) were the end point. Those ECG findings were relatively common and covered the majority (80 to 90%) of all abnormalities found in the general population before excluding subjects with symptomatic heart disease. Most ECG findings on most occasions were associated with an excess mortality from the three end-points in both men and women and among relatively young (age 30-49) and mature (age 50-69) adults. The strongest predictor of fatal events were Q-QS items and blocks. The most consistent predictors were ST-T findings, although this was true for men and not for women. Relative risk against the absence of abnormalities (one by one and all together) were adjusted by multivariate analysis feeding in the models some possible confounders, i.e. age, systolic blood pressure, serum cholesterol, cigarette consumption and body mass index. Relative risks in cells with more than 20 events (cells being separately made by men, women, the 5 ECG findings categories and the 3 end-points) were ranging 1.00 to 9.88 for Q-QS abnormalities, 1.03 to 3.76 for ST-T abnormalities, 1.28 to 5.14 for high R waves, 0.81 to 2.28 for arrhythmias and 0.79 to 3.59 for blocks. Most of these relative risks were statistically significant. Combinations of clinically relevant ECG findings in the same individual (LVH, possible and definite myocardial infarction) were rare but carried a severe prognosis with high and statistically significant relative risks among men (ranging between 3.19 and 7.24) while they could not be properly tested in most cells for women due to the small numbers involved. It is concluded that in the general population high rates of prevalent ignored ECG abnormalities in asymptomatic subjects are associated with significant excess mortality from CHD, CVD and all-cause mortality, suggesting a high prevalence of silent heart disease. PMID- 9199943 TI - Effects of bamiphylline on exercise testing in patients with syndrome X. AB - An abnormal stimulation of adenosine A1-receptors has been suggested to play a role in the pathogenesis of both chest pain and ischemia-like electrocardiographic changes in patients with syndrome X and a nonselective adenosine antagonist (theophylline) has been reported to be beneficial in these patients. In this study we investigated the acute effects of bamiphylline, a specific A1-receptor antagonist, in 16 patients with syndrome X (14 women, age 57 +/- 6 years), with both angina and ST-segment depression inducible during exercise testing. All patients underwent two treadmill exercise tests (Bruce modified protocol) on 2 separate days, 5 minutes after the end of randomized intravenous infusion of either placebo (saline solution) or bamiphylline (300 mg). Severity of chest pain was assessed by a 100 mm visual analogic scale. There were no significant differences in resting heart rate and blood pressure after bamiphylline or placebo. Rate-pressure product (20 600 +/- 5000 vs 20 200 +/- 5200 bpm.mmHg), time to 1 mm ST depression (549 +/- 196 vs 581 +/- 201 sec), time to angina (519 +/- 209 vs 571 +/- 196 sec), and exercise duration (717 +/- 134 vs 676 +/- 166 sec) were also not significantly different after bamiphylline or placebo, but there was a mild reduction of the severity of exercise-induced chest pain (30 +/- 22 vs 39 +/- 20 mm, p < 0.05) with the active drug. Thus, in patients with syndrome X, bamiphylline does not improve exercise-induced ST changes, suggesting that A1-receptors are not significantly involved in their appearance. In addition, bamiphylline had little effect on anginal pain, suggesting that this cannot be mediated exclusively by A1-receptor stimulation in these patients. PMID- 9199944 TI - Cardiac arrest during dobutamine stress echocardiography. AB - In this report, we describe the case of a woman with normal coronary arteries who experienced a cardiac arrest during a dobutamine stress test. The patient was successfully treated with external cardiac massage. The possible mechanisms underlying this unusual life-threatening side effect of dobutamine infusion are discussed. PMID- 9199945 TI - Autoimmunity in myocarditis and dilated cardiomyopathy: cardiac autoantibody frequency and clinical correlates in a patient series from Italy. AB - BACKGROUND: Organ- and disease-specific cardiac autoantibodies, detected by indirect immunofluorescence, represent markers of autoimmunity in a subgroup (25 35%) of patients with dilated cardiomyopathy or myocarditis from Northern Europe and the United States of America. Autoantibody frequencies, as well as associations between clinical and immunological features, may vary in patients from different countries, due to ethnically related differences in genetic susceptibility to autoimmune disease. METHODS: We assessed the frequency of cardiac autoantibodies in a series from Italy, including 91 subjects with idiopathic dilated cardiomyopathy (61 male, aged 49 +/- 11 years) and 11 with biopsy-proven (Dallas criteria) myocarditis (7 male, aged 23 +/- 16), including 2 cases of giant cell myocarditis. Controls were 160 patients with other cardiac disease, 141 with ischemic heart failure and 270 normals Cardiac antibody test was performed blindly by indirect immunofluorescence on normal human myocardium and skeletal muscle. RESULTS: The frequency of organ-specific cardiac autoantibodies was higher (p = 0.0001) in myocarditis (45%) and in dilated cardiomyopathy (20%) than in other cardiac disease (1%), in ischemic heart failure (1%), or in normals (2.5%). Cross-reactive antibodies were detected in similar proportions of study patients and controls. Both patients with giant cell myocarditis were antibody positive. Myocarditis patients with cardiac antibodies had shorter duration of symptoms compared to those who were antibody negative (0.4 +/- 0.3 vs 4 +/- 1 months, p = 0.004). In dilated cardiomyopathy, antibody status was not associated with any clinical or diagnostic feature. CONCLUSIONS: Autoimmunity is involved in a subset of patients with myocarditis and with dilated cardiomyopathy, regardless of their geographical origin or immunogenetic background. The antibody frequency in our dilated cardiomyopathy series from Italy tended to be lower than in other countries. This may reflect reduced antibody levels with disease progression and/or the recognised feature that Mediterranean populations are often less susceptible to autoimmune disease. PMID- 9199946 TI - Circadian variations of ventricular tachyarrhythmias detected by the implantable cardioverter-defibrillator. AB - OBJECTIVE: The purpose of this study was to prospectively evaluate the daily distribution of episodes of ventricular tachyarrhythmias triggering a last generation ICD implanted in survivors of cardiac arrest not receiving any antiarrhythmic drug. BACKGROUND: Previous studies reported a circadian variation of out-of-hospital sudden cardiac arrest. Survivors of cardiac arrest and patients with sustained ventricular tachyarrhythmias have a higher risk of recurrence, but there is a lack of detailed information on the daily distribution of ventricular arrhythmia capable of precipitating the recurrence of a malignant ventricular event. METHODS: We used the data stored by a last-generation implantable cardioverter-defibrillator (ICD) to prospectively evaluate circadian distribution of ventricular tachyarrhythmias in 30 survivors of cardiac arrest, implanted with an ICD during an antiarrhythmic drug-free period. RESULTS: During a follow-up of 475 +/- 127 days, a total of 571 ventricular arrhythmias was recorded. Of these stored events, 39 episodes were ventricular fibrillation (VF), 428 events were ventricular tachycardia (VT) and the remaining 104 device activations were VTs spontaneously terminating before device therapy. A circadian variation (p < 0.001) of episodes of ventricular tachyarrhythmias was evident. The incidence of ventricular arrhythmias sharply increased at 6 a.m. and reached a maximum 4 hours later, after which there was a short decline and then a small peak between 3 p.m. and 5 p.m. With respect to different arrhythmias, VF or VTs with a cycle length < or = 350 msec demonstrated a circadian variation. Instead, VTs with a cycle length longer than 350 msec or spontaneously terminating before device discharge had a more even distribution throughout the day. In addition, most of the VF or VTs with a cycle length < or = 350 msec occurred within a few hours after awakening, which was not the case for VTs with a cycle length longer than 350 msec. CONCLUSIONS: The data of this study clearly show the existence of different circadian variations in the occurrence of ventricular tachyarrhythmias, suggesting complex interactions between the autonomic nervous system, ventricular electrophysiological properties and the onset of arrhythmia. PMID- 9199947 TI - On-line three-dimensional intracoronary ultrasound for guidance of catheter based interventions. AB - Intracoronary ultrasound (ICUS) provides valuable information on the distribution and composition of atherosclerotic plaque. For this reason its use for guidance of interventional procedures has been advocated. Recently, on-line systems of three-dimensional reconstruction have been introduced and offer great potential for guidance of interventional procedures since valuable details on longitudinal architecture of the plaque under treatment are obtained. In this article we review the current clinical application of three-dimensional (3-D) ICUS and report our experience with the use of an on-line 3-D ICUS system for guidance of interventional procedures. In our experience 3-D reconstruction of ICUS proved to be a feasible method facilitating device selection and guidance of catheter based interventions. PMID- 9199948 TI - The "proximal isovelocity surface area" method in assessing mitral valve area in patients with mitral stenosis and associated aortic regurgitation. AB - BACKGROUND: This study compares the mitral valve area determined by Doppler color mapping of the proximal isovelocity surface area (PISA) and by Doppler pressure half-time with that obtained by two-dimensional planimetry in patients affected by mitral stenosis, with and without associated aortic regurgitation. Pressure half-time frequently overestimates the mitral valve area in patients with mitral stenosis and associated aortic regurgitation. PISA is an alternative method for determining mitral valve area in mitral stenosis and is not influenced by regurgitant lesions. METHODS: We studied 76 patients with mitral stenosis; aortic regurgitation > or = 2 was present in 24 patients. The PISA was recorded from the apex and the transmitral maximal flow rate, Q (ml/s), was calculated using the hemispheric equation Q = 2 pi R2 x AV x alpha/180, where R (cm) is the maximal radius of the PISA, AV (cm/s) is the aliasing velocity and alpha/180 is a correction factor accounting for the alpha inflow angle formed by the mitral leaflets. Mitral valve area, A (cm2), was calculated by continuity equation A = Q/V, where V (cm/s) is the peak transmitral flow velocity measured by continuous wave Doppler. RESULTS: The mitral valve area by two-dimensional planimetry (range 0.5-2.4 cm2; mean 1.33 +/- 0.41 cm2) was consistent with both PISA (r = 0.83; SEE 0.23 cm2) and pressure half-time (r = 0.79; SEE 0.25 cm2) methods. Similar agreement was found for the 36 patients with mitral regurgitation and for the 30 patients in atrial fibrillation. However, in patients with aortic regurgitation > or = 2, pressure half-time overestimated two-dimensional and PISA determined mitral valve areas by 0.24 +/- 0.25 cm2 (p < 0.01). CONCLUSIONS: In patients with mitral stenosis and significant aortic regurgitation, the PISA method is more accurate than pressure half-time in assessing mitral valve area. This method may be a reliable alternative when pressure half-time is affected by aortic regurgitation and two-dimensional planimetry images are unsuitable for anatomic evaluation. PMID- 9199949 TI - Ten-year experience with endomyocardial biopsy in myocarditis presenting with congestive heart failure: frequency, pathologic characteristics, treatment and follow-up. AB - The present study summarizes our ten-year (1985-1995) experience with endomyocardial biopsy (EMB) in patients with idiopathic congestive heart failure (CHF), with specific reference to frequency of myocarditis, treatment policy, relative benefits, and follow-up. Of the 601 patients who constituted our series, 38 were clinically suspected of having myocarditis on the bases of a very recent onset of congestive heart failure and/or of arrhythmias and/or of conduction disturbances, and of a close-to-recent history of flu-like febrile illness. Corresponding EMBs showed myocarditis in 16 of the 38 cases (42.1%). A further 10 EMBs, from patients with a recent onset of congestive heart failure without prior infection episodes, showed myocarditis. Therefore, biopsy-proven myocarditis occurred in 26 of the 601 patients (4.3%). Of the 26 cases, 21 were lymphocytic, 1 was necrotizing granulomatous, 1 was eosinophilic and occurred in a patient who later developed overt zoonosis, 1 had some giant cells within endocardial inflammatory infiltrates, and 2 were borderline forms. In active myocarditis, inflammatory cells mostly constituted of T-lymphocytes (CD45RO+) with sparse macrophages (CD68+) and a few B cells (CD20+). B-lymphocytes and macrophages, along with activated T-lymphocytes, all expressed MHC class II HLA DR molecules, which were also expressed "de novo" by activated endothelial calls of capillaries and of small intramural vessels. HLA DR revealed itself as a very useful marker for the detection of activated inflammatory and endothelial cells. We also noted an increase in the number of perivascular and interstitial mast cells. Ultrastructural study was helpful for the characterization of myocyte damage and of interactions between inflammatory cells and myocytes. In 4 cases (1 of whom was later revealed as HIV positive, and subsequently died of AIDS), we found microreticulotubular structures in endothelial cells of small vessel and capillaries; in 7 cases, there were myocyte changes similar to those described in polymyositis; in 1 case, we observed subplasmalemmal buddings, but no viral particles; in 6 cases, there was extensive myocyte damage with myofibrillar lysis and focal adipous metaplasia; the remaining 6 cases showed myocyte damage of differing extent and severity; in the borderline forms, such damage coexisted with interstitial fibrosis. One of the 21 lymphocytic myocardites was not treated because during hospital screening the patient proved to be HIV positive; of the remaining 20 active myocardites, 11 were treated with a 6-month tapered steroid and azathioprine protocol (one was treated for 24 months), while 9 were not treated. The corresponding follow-up was: 6 deaths (congestive heart failure), 2 cardiac transplants and 3 survivals (1 with pace-maker) in the treated group, and 3 deaths (2 of congestive heart failure and 1 of sudden death), 1 cardiac transplant and 5 survivals (1 on the waiting list for transplantation) in the non treated group. One of the 2 patients with borderline myocarditis died of congestive heart failure, and 1 is alive. Of the 22 patients with clinical diagnosis of myocarditis and negative biopsy, 7 died of congestive heart failure (2 on the waiting list for transplantation), 4 underwent cardiac transplantation, and 11 are alive (1 is awaiting transplantation). Of the 20 patients currently alive, 1 was originally in NYHA class III, 15 were in class II and 4 were in class I. Of the 20 overall patients who died, 12 were originally in NYHA class IV, 6 in class III, 2 in class II; of the 8 patients who underwent transplantation, 6 were originally in NYHA class IV and 2 in class III. Our overall experience shows that the frequency of myocarditis diagnosed according to Dallas criteria is high in patients with clinical diagnosis of myocarditis, while it is extremely low in dilated cardiomyopathy patients. This finding suggests that, although non-specific, recent onset of symptoms and prior febrile infe PMID- 9199950 TI - Echocardiographic evaluation of systolic and diastolic left ventricular function following arterial switch operation in the neonatal period for transposition of the great arteries. Midterm results. AB - BACKGROUND: Neonatal arterial one-stage switch operation (ASO) for transposition of the great arteries (TGA) is currently the procedure of choice for TGA. There is a potential risk of myocardial damage related to coronary artery reimplantation and sudden pressure overload imposed on the left ventricle after discontinuation of the cardiopulmonary bypass. OBJECTIVES: This study was carried out in order to evaluate left ventricular systolic and diastolic function in children following ASO. METHODS: We studied 32 children (22 M, 10 F), mean age 23.7 +/- 24.6 months (range 0.5-97.2) following ASO, without any hemodynamically significant residual stenosis by 2D- and Doppler-echocardiography. Twenty-five had TGA with intact ventricular septum and 7 with ventricular septal defect. Mean age at time of one-stage repair was 8.9 +/- 6.9 days (range 2-30) and the mean time of follow-up 23.5 +/- 24.6 months (range 0.3-96). At the time of evaluation all the children were asymptomatic. Regional wall motion of the left ventricle was assessed by 2D-echo. Volumes, mass index, M/V ratio, afterload, systolic function (FS% and VCFc), contractility (stress-velocity SVI and stress-shortening SSI relations) and preload (functional preload index FPI = SSI-SVI) of the left ventricle were determined by m-mode echo together with non-invasive blood pressure monitoring. Left ventricular diastolic function was determined by PW Doppler of transmitral flow. All parameters were compared with those of 32 normal controls matched for age, sex and body surface area. RESULTS: Left ventricular regional wall motion was normal in all but 5 cases who showed a slight reduction of the septal systolic motion. Volumes and EF% did not differ in post-ASO group vs controls. There was a small increase of the mass index in the post-ASO group vs controls (62.8 +/- 14.1 vs 56.2 +/- 6.5 g/m2; p = 0.02) and the M/V ratio did not differ. FS% and VCFc were not different between the 2 groups. Peak and end systolic meridional stress also did not differ in the 2 groups. A normal contractile state was present in the post-ASO group. The preload was slightly reduced in the post-ASO children (FPI -0.6 +/- 0.94 vs 0.07 +/- 0.63; p = 0.001). Parameters of left ventricular diastolic function were not significantly different between the 2 groups. CONCLUSIONS: In conclusion in children undergoing neonatal ASO for TGA with intact septum or ventricular septal defect, evaluated after a mean post-surgical follow-up of 2 years, systolic and diastolic performance of the left ventricle was normal. Regional wall motion abnormalities with slightly reduced septal motion were detected in 5 cases. The reason for the small increase in mass index is unknown. The slight reduction of the preload is related to the routine drug therapy in the patients studied early after surgical repair. PMID- 9199951 TI - Is quantitative angiography sufficient to guide stent implantation? A comparison with three-dimensional reconstruction of intracoronary ultrasound images. AB - Intracoronary ultrasound (ICUS) frequently reveals stent underexpansion despite a satisfactory angiographic result by visual assessment. Whether on-line quantitative coronary angiography (QCA) alone can guide optimal stent deployment is still unknown. The aim of the study was to assess the usefulness of quantitative coronary angiography in the evaluation of optimal stent expansion, confirmed with a new on-line system of 3-D reconstruction of ICUS. The results obtained with 3-D ICUS were compared with the measurements achieved with QCA analyses in 49 patients (70 stents: 31 Palmaz-Schatz, 22 Wallstent, 7 Cordis, 6 Micro-stent, 2 Gianturco-Roubin, 2 Multi-Link). Following delivery of the stent, high pressure intrastent balloon inflation (14.2 +/- 3.3 atmospheres) was performed in all 70 stents. Optimal stent implantation by QCA was defined as minimal lumen diameter post-stenting > or = 90% of the reference diameter preintervention. Percent diameter stenosis (% DS) post-stenting was defined as minimal lumen diameter divided by the reference diameter post-stenting. The on line 3-D ICUS reconstructions and measurements were performed processing the images on-line in the catheterization laboratory with an automated contour detection algorithm based on acoustic quantification. ICUS criteria for optimal stent expansion were defined as: 1) complete apposition of stent struts to vessel wall; 2) minimal stent lumen cross-sectional area > or = 80% of the average lumen area of the proximal and distal reference segments; 3) symmetry index (minimum divided by maximum lumen diameter) > 0.7. Ninety-seven percent of the deployed stents met the QCA criteria. Whilst 3-D ICUS documented complete stent apposition to the vessel wall in all cases and a symmetric expansion in 65 of 70 lesions (93%), the stent minimal lumen area was > or = 80% only in 30 out of 70 stents (43%). The diagnostic sensitivity and specificity at 10% residual diameter stenosis provided by QCA for optimal stent expansion compared to 3-D ICUS criteria were 86 and 45%, respectively. In conclusion 3-D ICUS criteria of adequate stent expansion were achieved only in 43% of patients despite the application of aggressive strategies of stent deployment leading to optimal results with quantitative angiography. Ten percent residual diameter stenosis provided by QCA may be an acceptable alternative for optimal stent deployment in clinical practice. The clinical benefit of an ICUS guided approach of stent deployment and of a lower cost strategy using on-line QCA guidance should be compared in large prospective randomized studies. PMID- 9199952 TI - Identification of viable myocardium early after acute myocardial infarction under beta-blockade by enoximone echocardiography. AB - The influence of the beta-blocker metoprolol on the capacity either of low-dose dobutamine echocardiography or the recently introduced enoximone echocardiography to detect viable dysfunctioning myocardium after myocardial infarction was investigated. Initial clinical experience would suggest that the phosphodiesterase III inhibitor enoximona could be an alternative pharmacological stimulation, inducing an increase in contractility in the presence or absence of beta-receptor stimulation. Ten patients with a baseline low-dose dobutamine echocardiographic test (up to 10 micrograms/kg/min) positive for myocardial viability in > or = 1 segment(s), performed 4-5 days after a first acute myocardial infarction treated with rtPA, were randomized after the administration of intravenous metoprolol (15 mg in three 5-mg boluses) either to dobutamine (up to 15 micrograms/kg/min) or to an enoximone intravenous bolus (1 mg/kg over 5 min) under echocardiographic monitoring, in a crossover sequence, with a 24-h interval. The infarct related artery was patent (TIMI grade 2 o 3) in all the patients. Follow-up echocardiograms were performed 5-7 weeks later. Resting asynergy was found in 40 segments; of these, 17 were viable. All the viable segments remained unresponsive during the post-metoprolol dobutamine infusion, while improved their contractility during enoximone echocardiography. Two patients suffering from early post-infarction angina underwent coronary angioplasty successfully. Eight out of ten patients (2 revascularized and 6 not) showed contractile recovery in a total of 14 segments at the follow-up echocardiogram. Sensitivity, specificity and overall accuracy in predicting reversible dysfunction after acute myocardial infarction for enoximone echocardiography were 93, 85, and 88%, respectively. Our results support the value of enoximone echocardiography in the identification of myocardial viability after myocardial infarction, in patients treated with beta-blockers, which interfere heavily with the results of dobutamine echocardiography. PMID- 9199953 TI - Prevalence and correlates of echocardiographic determined left ventricular hypertrophy in 2318 asymptomatic middle-aged men: the ECCIS project. Epidemiolgia e Clinica della Cardiopatia Ischemica Silente. AB - It is well established that left ventricular hypertrophy is a strong and independent risk factor for cardiovascular morbidity and mortality. This study was designed to determine the prevalence and correlates of left ventricular hypertrophy (LVH) among a sample population of 2318 totally asymptomatic men aged 40-59. This sample is a subset of the participants in the ECCIS Project. Left ventricular mass was estimated by echocardiography. The following individual variables were employed in the multiple linear regression analyses: age, diastolic and systolic blood pressure at rest and at peak exercise, body mass index, body surface area, conditioning physical activity. Three indexes of left ventricular mass were used: left ventricular mass/height, left ventricular mass/body surface area and "adjusted left ventricular mass" derived from adjustment, using a regression model, of left ventricular mass by age, body mass index and body surface area. The sample was subdivided in 3 blood pressure classes; normotensive (n = 1605), borderline (n = 390) and hypertensive (n = 323). All the variables considered with the exception of diastolic blood pressure both at rest and peak exercise were significantly correlated with left ventricular mass. Upper normal limits for left ventricular mass indexed to height and body surface area and of adjusted left ventricular mass were 143 g/m, 129 g/m2, and 245 g respectively. The prevalences of left ventricular hypertrophy, as determined by the reference standard of left ventricular mass/height, left.ventricular mass/body surface area and adjusted left ventricular mass, ranged 2.7-3.2% in the normotensive group, 4.2-5.4% in the borderline group and 11.8-14.5% in the hypertensive group, and were lower using adjusted left ventricular mass index. The results of this study show that the prevalence of left ventricular hypertrophy using adjustment by age, body surface area and body mass index reduces variability of left ventricular mass associated with age and body size and may be useful for the correct identification of left ventricular hypertrophy and hypertensive heart disease. PMID- 9199954 TI - Sleep disorders and breathing alterations in patients with chronic heart failure. AB - Cheyne-Stokes respiration can appear during sleep in patients with chronic heart failure and is generally attributed to a tendency to hyperventilate causing PCO2 to fall below the apnea threshold. We recorded sleep pattern and nocturnal arterial oxygen desaturation during Cheyne-Stokes respiration and correlated those with hemodynamic alterations, in order to investigate their possible role in the evolution of chronic heart failure. Sixty chronic heart failure patients, after optimization of therapy, underwent a polysomnographic study and hemodynamic and echocardiographic evaluations within a few days. The patients were then enrolled in the follow-up of our pre-transplantation program. Only slight alterations of sleep architecture were detected. During sleep, Cheyne-Stoke respiration was present in 50% and arterial oxygen desaturations in 54% of patients. An increased pulmonary wedge pressure (24.7 +/- 8.3 vs 16.7 +/- 8.9 mmHg, p < 0.000) was significantly correlated with the presence of nocturnal Cheyne-Stokes episodes, while cardiac index was not (1.9 +/- 0.6 vs 2.0 +/- 0.5 l m-2 min-1, p = 0.42). In a multivariate analysis of hemodynamic and polysomnographic data, mortality or heart transplantation in status 1 was predicted at the two year follow-up only by an increased pulmonary wedge pressure. In conclusion, in advanced chronic heart failure, with optimized therapy, nocturnal Cheyne-Stokes respiration is present in half of the cases, with concomitant falls in arterial oxygen desaturation. These events were not independently predictive of mortality. The strong correlation found between increased left ventricular filling pressure and presence of Cheyne Stokes respiration and the lack of correlation with cardiac index suggest that other hemodynamic mechanisms besides reduced cardiac output are responsible for this respiratory abnormality. PMID- 9199955 TI - Restrictive cardiomyopathy due to desmin accumulation in a family with evidence of autosomal dominant inheritance. AB - OBJECTIVE: A familial case of restrictive cardiomyopathy due to desmin accumulation characterized by severe disturbances of cardiac conduction is described. BACKGROUND: Desmin is an intermediate filament normally present in the myocardium, particularly in the Purkinje fibres, in the skeletal and in the smooth muscle. METHODS: Resting electrocardiogram, 2-dimensional and Doppler echocardiogram, cardiac catheterization, electrophysiological study have been performed in all siblings. Informed consent for endomyocardial biopsy was obtained only in one patient. RESULTS: The mother showed bilateral pes cavus and complained of episodes of vertigo at the age of 36 years. At that time she was submitted to electrophysiological study and to permanent pacing. After 15 years of good health conditions, she developed heart failure and underwent cardiac transplantation. A 21 year old son had a syncope; his ECG was similar to that of his mother; a permanent pacemaker was implanted and a diagnosis of restrictive cardiomyopathy with desmin accumulation was confirmed at histopathology study. Afterwards, another 24 year old sib had a syncope with head trauma: ECG showed right atrial enlargement, left bundle branch block. After electrophysiological study, he started antiarrhythmic therapy. This patient showed bilateral pes cavus. CONCLUSIONS: The early manifestation of desmin accumulation may be intraventricular conduction disorders that can be often controlled by pacemaker implantation. Clinical symptoms of heart failure may be absent for a long period of time. Pedigree analysis is most consistent of autosomal dominant inheritance. PMID- 9199956 TI - Early results of minimally invasive aortic valve replacement. Experience with the first 34 cases. AB - BACKGROUND: The method of replacing the aortic valve via a minithoracotomy has been reported in the recent literature. This strategy has clear advantages. However, further refinements of the process make the procedure even less invasive. METHODS: Aortic valve replacement was performed in 34 patients whose age ranged from 49 to 82 years, averaging 69 +/- 8 years. As access route, a right parasternal minithoracotomy about eight cm long and without rib resection was used. Cardiopulmonary bypass was connected through the same access. The standard surgical technique and equipment were employed. RESULTS: There were neither intraoperative complications nor hospital death. All patients, except two could be discharged home within one week. Cardiopulmonary bypass time, aortic cross-clamp time, and total operation time averaged 110 +/- 25, 73 +/- 19, and 183 +/- 38 minutes, respectively. Three patients could be extubated in the operating theater, and the others on the intensive care units at an average of 9 +/- 7 hours postoperatively. One patient had to be re-entered immediately after extubation because of a bleeding from the aortic cannulation site. A second patient, who was initially operated because of a florid aortitis, had a limited periprosthetic leak two months postoperatively which was repaired thereafter. CONCLUSIONS: The advantages of the present method include further reduction of surgical trauma, preservation of chest wall integrity, early mobilization, recovery and rehabilitation of the patient. Improvements in the surgical technique include avoidance of groin cannulation, simpler equipment, and an easy access through a mid-sternotomy in case of reoperation. PMID- 9199957 TI - Mobile high-efficiency-filter air cleaners. AB - Mobile high-efficiency-filter air cleaners (MHEFACs) are freestanding air filtration and recirculation systems used to supplement poor ventilation or to compensate for a lack of ventilation in general-use and high-risk areas of healthcare facilities. These devices are primarily used as an engineering control to reduce the concentration of infectious tuberculosis (TB) droplet nuclei in the room air. This Update supplements our October 1995 Evaluation (Health Devices 24[10]) of these devices, in which we evaluated 14 MHEFACs from 10 manufacturers. In the current Update, we evaluate 3 additional MHEFACs from 2 additional manufacturers. We also present update information for the previously evaluated units. Unless otherwise specified, we tested the new units against the same criteria and using the same test methods as those detailed in our October 1995 Evaluation. For brevity, we have not reprinted criteria or test methods that were identical to those used in the original study. In the introduction below, we briefly describe the purpose and use of MHEFACs, focusing on the application for which they are primarily intended: controlling the spread of TB. For more detailed information about this technology, the environments in which MHEFACs are used, and the factors to consider when selecting this type of device, we encourage readers to refer to the following items in the October 1995 issue: The Clinical Perspective, "The Resurgence of Tuberculosis-From Old Problem to New Menace." The Clinical and Technical Overview, in which we discuss the mechanisms of airborne infection and describe and compare some of the controls used to reduce the incidence of such infections. The Guidance Section, "Issues in Selecting, Purchasing, and Using Mobile High-Efficiency-Filter Air Cleaners." The supplementary articles, "The Wells-Riley Equation," "Airborne infection Control Terms and Abbreviations," and "Air Mixing and Airflow Rates." Descriptions of the newly evaluated units, along with our test results and ratings, are presented in individual Product Profiles following the introduction. Following the last profile, we present a comprehensive Conclusions section that details how the newly evaluated units compare with the previously evaluated ones. PMID- 9199958 TI - Trends in medical equipment service. AB - Driven by the recent emphasis on cost containment in the healthcare industry as a whole, hospitals, manufacturers, and service providers have been spurred to try to find ways to reduce their costs and increase their revenues. As a result of the measures taken to achieve these goals, the medical equipment service industry has been experiencing a period of rapid change. In this environment, traditional alternatives for servicing medical equipment are being challenged, and new alternatives are being developed. The changes occurring in this industry are of vital interest to clinical engineers and technicians because it is those changes that will determine the future role of these professionals in the hospital. While such technology experts will not become obsolete-the reliance on medical technology to provide optimal healthcare will certainly continue in the United States and throughout the rest of the developed world-the roles of these experts could very likely change. As hospitals focus more narrowly on their core functions, noncore functions such as servicing and managing technology will come under scrutiny as possible candidates for outsourcing to reduce the hospital's administrative responsibilities and cut its cost burden. In this article, we review the recent past of the medical equipment service industry and describe some current trends that will play a role in shaping the future of this industry. We also discuss how the changes currently taking place will likely affect the viability of in-house clinical engineering departments. In our discussion, we spotlight service issues related to radiology equipment because (1) this equipment dominates service expenditures and (2) the servicing of these devices has spawned some of the changes causing hospitals to reexamine how all their medical equipment is being serviced and managed. PMID- 9199959 TI - Proper use and inspection of patient lifts. PMID- 9199960 TI - Intermittent loss of signal from Hewlett-Packard Component Monitoring System (CMS) modules. PMID- 9199961 TI - Loss of function and blanking of the display on Hewlett-Packard M2350A and M2360A central station monitors. PMID- 9199962 TI - Automatic shutdown of Ohmeda pulse oximeters. PMID- 9199963 TI - The IgV domain of human B7-2 (CD86) is sufficient to co-stimulate T lymphocytes and induce cytokine secretion. AB - B7-1 (CD80) and B7-2 (CD86) are genetically and structurally related molecules expressed on antigen-presenting cells. Both bind CD28 to co-stimulate T lymphocytes, resulting in proliferation and cytokine production. The extracellular portions of B7-1 and B7-2 which bind to CD28 and CTLA-4 are related to Ig variable (V) and Ig constant (C) domain sequences. Recent reports have described splice variant forms of B7 proteins which occur in vivo and are of unknown function. Here we describe soluble recombinant forms of B7-1 and B7-2 containing either both of the Ig-like extracellular domains or the individual IgV or IgC domains coupled to an Ig Fc tail. Soluble B7-1 and B7-2 bind to CD28 and CTLA-4, and effectively co-stimulate T lymphocytes resulting in their proliferation and the secretion of cytokines. Furthermore, the IgV domain of B7-2 binds CD28 and CTLA-4, competes with B7-1 and B7-2 for binding to these receptors, and co-stimulates T lymphocytes. Cross-linked soluble B7-2v was the most potent co-stimulatory molecule tested and was active at a concentration approximately 100-fold lower than cross-linked soluble B7-1 or B7-2 proteins. When bound to tosyl-activated beads, B7-2v was capable of sustaining multiple rounds of T cell expansion. These data complement the description of naturally occurring variants to suggest that T cell co-stimulation in vivo may be regulated by soluble or truncated forms of B7 proteins. PMID- 9199964 TI - A broad cytotoxic T lymphocyte response to influenza type B virus presented by multiple HLA molecules. AB - The HLA restriction and epitope specificity of cytotoxic T lymphocytes (CTL) involved in recovery from influenza type B infection have not been extensively characterized. Here lymphocytes obtained from a healthy individual contained virus-specific CTL restricted by class I HLA molecules, HLA-A1, A2, B7 and B8, and the class II HLA molecules, HLA-DR1 and DR3. Four conserved viral epitopes were predicted from allele-specific motifs for peptides interacting with HLA-B8 and HLA-DR1. Bulk CTL recognized three 9mer HLA-B8-restricted peptides from nucleoprotein, residues 30-38, 263-271 and 413-421, and a 13mer HLA-DR1 restricted peptide from hemagglutinin, residues 308-320. The epitopes presented by HLA-A1, HLA-B7 and HLA-DR3 remain undefined. Peptide-specific CTL lines recognized influenza type B virus-infected cells indicating the peptides are representative of naturally processed epitopes. A hemagglutinin peptide-specific CD4 CTL clone expressed approximately 200 molecules of perforin mRNA/cell, suggestive of a functional perforin pathway for target cell lysis. The results indicate a broad CTL response composed of both CD8 CTL and CD4 CTL recognizing viral epitopes presented by multiple HLA molecules. PMID- 9199965 TI - Human T cell responses against the major cysteine proteinase (cruzipain) of Trypanosoma cruzi: role of the multifunctional alpha 2-macroglobulin receptor in antigen presentation by monocytes. AB - Chagas' disease patients (CDP) develop both humoral and cellular immune responses against the major cysteine proteinase (cruzipain) from Trypanosoma cruzi. Here we demonstrate that complexes formed by cruzipain and alpha 2-macroglobulin (alpha 2M) are efficiently internalized by human monocytes, and that this process results in enhanced presentation of cruzipain peptides to CD4+ T cells from CDP. Purified or serum alpha 2M binds to polymorphic cruzipains, but only a fraction of the proteinases become covalently linked. Once bound to alpha 2M, fluorescein labeled cruzipain (FITC-cruzipain) or [125I]cruzipain were more efficiently internalized by normal peripheral blood mononuclear cells (PBMC) or monocytes; this effect was abolished by (I) pre-treating the cells with receptor-associated protein (rRAP), a known antagonist the of alpha 2M receptor (alpha 2MR/LRP), and (II) inactivating [125I]cruzipain's active site prior to the reaction with alpha 2M, indicating that the exposure of receptor binding sites on alpha 2M complexes required bait region cleavage. We then sought to determine if the alpha 2MR/LRP dependent uptake of alpha 2M:cruzipain by monocytes resulted in increased CD4+ T cell responses of PBMC-CDP (n = 13). These effects were only revealed after depletion of CD19+ B lymphocytes from PBMC-CDP; the threshold of T cell stimulation was far lower in cultures stimulated with alpha 2M:cruzipain, as compared to antigen alone. Myocardial specimens from CDP with chronic myocardiopathy (three necropsies) were analyzed by immunohistochemistry with mAb anti-cruzipain or anti-alpha 2MR/LRP (CD81+). Extracellular depots of cruzipain were localized amidst inflammatory mononuclear infiltrates, part of which contained CD91+ macrophage-like cells. Ongoing studies should clarify if T. cruzi cysteinyl proteinases play a role in the pathogenesis of Chagas' heart disease. PMID- 9199966 TI - HIV type 1 Tat protein enhances activation-but not Fas (CD95)-induced peripheral blood T cell apoptosis in healthy individuals. AB - T cell apoptosis may play an important role in the depletion and functional defects of T cells in HIV disease. A number of investigators have shown that peripheral blood T cells in HIV disease undergo spontaneous and activation induced apoptosis. We found recently that peripheral blood T cells from HIV+ individuals undergo apoptosis when stimulated through Fas. Also, a number of investigators have shown that Tat protein from HIV-1 can increase spontaneous and activation-induced apoptosis. In the present study we examined the effect of HIV type 1 Tat protein on spontaneous, activation-induced and Fas-induced apoptosis of peripheral blood T cells from HIV- individuals. We find that Tat protein has no effect on spontaneous apoptosis but does enhance activation-induced apoptosis of both CD4+ and CD8+ T cells. Tat, however, failed to enhance Fas-induced apoptosis of CD4+ and CD8+ T cells. Examining the mechanisms by which Tat induces apoptosis, we found that inhibitors of reactive oxygen intermediate (ROI) generation or neutralizers of ROI, such as rotenone, a potent inhibitor of mitochondrial complex I of the respiratory chain, and 3,3,5,5 tetramethylpyrroline N-oxide (TMPO), an electron spin trap, could both enhance the spontaneous apoptosis induced by Tat. This enhancement of Tat-induced apoptosis by rotenone and TMPO was independent of ICE activation as it could not be inhibited by the tripeptide z-VAD-fmk, an irreversible inhibitor of ICE/ced-3 protease homologs. These findings suggest that Tat induced enhancement of activation-induced cell death may involve complex mechanisms, some of which are ROI independent. These results indicate that a HIV-specific mechanism other than Tat is responsible for the previously observed increased susceptibility of peripheral blood T cells from HIV-infected individuals to undergo apoptosis in response to Fas stimulation. PMID- 9199967 TI - CD4 and CD8 expressions in African green monkey helper T lymphocytes: implication for resistance to SIV infection. AB - We found that most peripheral CD4 cells co-express a low density of CD8 alpha antigen in African green monkeys (AGM). Further, the cell surface expression of CD4 and the expression of CD4 mRNA underwent a decrease when purified CD4CD8low cells were cultured with mitogen and IL-2. These observations suggest that AGM CD4 cells are subject to loss of CD4 expression after lymphocyte activation. Part of the peripheral CD8 fraction exhibited a significant helper activity which suggested the phenotypic conversion in helper T cells from CD4+ to CD4- in vivo. Simian immunodeficiency virus (SIV) grew well in CD4 panning cells following SIV infection. In contrast, CD4CD8low cells were resistant to SIV infection after their conversion to CD4- cells. PMID- 9199969 TI - IL-4 receptor complexes containing or lacking the gamma C chain are inhibited by an overlapping set of antagonistic IL-4 mutant proteins. AB - IL-4 signals on lymphocytes and other cells through a heterodimeric complex of two cytokine receptor proteins, IL-4R alpha and gamma C. IL-4 variants with mutations in the positions Tyr124, and (less efficiently) Arg121 and Ser125 are deficient in interaction with gamma zero and can be applied as IL-4 antagonists. Some cells respond to IL-4 without using gamma zero presumably by recruiting an alternative subunit into the receptor complex. We have investigated the effects of IL-4 mutant proteins on cells bearing this second type of IL-4 receptor complex. Mutations in the amino acid residues Arg 121 and Tyr124, but not Ser125, completely prevented cellular responses mediated by type 2 IL-4 receptors. Both receptor types are therefore affected by mutations in an overlapping but not identical site of the IL-4 molecule. PMID- 9199968 TI - Differential function of CD80- and CD86-transfected human melanoma cells in the presence of IL-12 and IFN-gamma. AB - Introduction of co-stimulatory molecules like CD80 and CD86 represents a means to augment the immunogenicity of tumor cells and to induce immune responses directed at tumor antigens. Here we compared CD80- and CD86-transfected human melanoma cells to induce primary immune responses by their capacity to promote proliferation of human allogeneic resting T lymphocytes. CD80- and CD86 transfected SkMel63 melanoma cells induced T cell activation to a comparable degree, which was found to be independent of the cell surface density of these co stimulatory molecules. Co-expression of CD80 and CD86 did not result in a synergistic increase in T cell proliferation. Both CD80 and CD86 transfectants induced the proliferation of isolated CD4+ or CD8+ T cells. Exogenous IL-2, IL-4 and tumor necrosis factor-alpha respectively enhanced primary T cell proliferation independent of CD80 or CD86 expression. Interestingly, differential activities of CD80 and CD86 were observed following stimulation of resting T cells in the presence of IL-12. Whereas IL-12 increased T cell proliferation in the presence of CD86-transfected melanoma cells, it exhibited an inhibitory function in the presence of CD80-expressing SkMel63 cells. Experimental evidence indicates that this inhibitory effect was mediated by IFN-gamma since (I) IFN gamma secretion of stimulated T cells was augmented by IL-12, (II) exogenous IFN gamma also inhibited T cell proliferation induced by CD80- but not CD86 transfected SkMel63 cells and (III) the inhibitory effect of IL-12 was blocked by an anti-IFN-gamma mAb. PMID- 9199970 TI - Molecular cloning of a pig homologue of membrane cofactor protein (CD46). AB - Organs of transgenic pigs that express human complement regulatory proteins are under assessment as an alternative to transplantation. A major barrier to the transplantation of pig organs is the hyperacute rejection caused by pre-existing antibodies and complement. Pig cells are very susceptible to human complement, presumably because pig cell-surface complement regulatory proteins are inefficient against it. Expression of human complement regulatory proteins, such as decay-accelerating factor and membrane cofactor proteins (MCP or CD46), by means of transgenes would confer resistance to human complement upon pig cells, thereby preventing hyperacute rejection. To express sufficient levels of human complement regulatory proteins at appropriate sites, regulatory elements of genes of pig membrane-bound complement regulatory proteins would be useful. To obtain their cDNAs, we transfected human cells with a pig cDNA library, selected cells by incubation with pig complement and rescued the plasmids. We cloned a cDNA for the pig homologue of MCP, pMCP. The cDNA encoded a predicted protein of 363 amino acids with 42% amino acid identity with human MCP. The pMCP consisted of four short consensus repeats, a Ser/Thr/Pro-rich domain, and transmembrane and cytoplasmic domains. Recombinant soluble pMCP that lacked transmembrane and cytoplasmic domains had factor I cofactor activity in C3b cleavage, indicating that it is functionally, as well as structurally homologous to MCP. FACS analysis with anti-pMCP mAb demonstrated that pMCP is expressed on all blood leukocytes, erythrocytes, and on endothelial and epithelial cell lines. PMID- 9199971 TI - Positive selection induces CD4 promoter and enhancer function. AB - Developmental expression of the CD4 gene in mature T cells is controlled by at least four transcriptional control elements: a promoter, two enhancers and a silencer. In this report we use a transgenic approach to study the mechanisms in which these elements interact to convey appropriate tissue- and cell-specific expression at all stages of T cell development. Our data indicate that the control of CD4 gene expression requires the interaction of multiple elements functioning in different combinations at different stages of T cell development. Expression of the CD4 gene in immature CD4+CD8+ thymocytes requires a third enhancer element located in the 3' flanking region of the CD4 gene. Interestingly, the CD4 promoter and proximal/distal enhancers first begin to function at the HSAlo CD69lo H-2Khi CD4 single-positive stage; cells of this phenotype are believed to have survived positive selection. These data indicate that the CD4 promoter and late enhancer elements are induced by positive selection; thus, the final maturation process is an active event that requires the initiation of a novel program of gene expression. PMID- 9199972 TI - Impaired antigen presentation by murine I-Ad class II MHC molecules expressed in normal and HLA-DM-defective human B cell lines. AB - The inability of certain antigen processing mutant cell lines to present intact proteins to T cells and to form SDS-stable MHC class II dimers has been shown to result from defective expression of HLA-encoded DMA and DMB genes. We have utilized some of these mutants to determine species compatibility of antigen presentation components. Mouse MHC class II I-Ad cDNA was transfected into the human B cell lymphoblastoid cell lines 8.1.6, 7.9.6 (a mutant cell line derived from 8.1.6) and an independent deletion mutant T2 (called 8.1.6d, 7.9.6d and T2.d respectively). These cells were than examined for various functions in antigen presentation. Interestingly, none of the cells transfected with I-Ad presented peptides derived from intact proteins to specific T cell hybridomas. However, presentation of synthetic peptides by these cells was normal. The ability to form SDS-stable dimers was dramatically reduced in the transfectants. In addition, I Ad molecules at the cell surface appeared loaded predominantly with the invariant chain peptides, CLIP. These properties of the I-Ad transfectants are identical to those described for HLA class II molecules expressed in HLA-DM mutants. Perhaps the most interesting finding was the inability of I-Ad in 8.1.6 to present protein antigens. Since 8.1.6 cells present antigens to HLA-DR, DP, DQ-restricted T cells and also have intact HLA-DM and invariant chain (II) functions, these results argue that some component of human antigen processing machinery is incompatible with I-Ad molecules. PMID- 9199973 TI - The thymus and self-tolerance: co-existence of encephalitogenic S100 beta specific T cells and their nominal autoantigen in the normal adult rat thymus. AB - The adoptive transfer of auto-reactive T cells specific for S100 beta protein mediates experimental autoimmune panencephalomyelitis, an inflammatory autoimmune disease of the nervous system and eye. However, unlike classical encephalitogenic autoantigens which are components of the myelin membrane and restricted to the nervous system, S100 beta is expressed by many different cell types in a wide variety of peripheral tissues. We now report that S100 beta is also expressed within the rat thymus from embryonic day 16 through to adulthood at which time point the protein is localized within stroma cells of the thymic medulla. However, despite the continued expression of this autoantigen within the thymic microenvironment it proved possible to isolate encephalitogenic, S100 beta specific CD4+ alpha beta TCR T cell lines from the naive adult rat thymus. These T cell lines were highly specific for S100 beta, and following activation in vitro and adoptive transfer initiate an inflammatory response in the central nervous system and eye of naive syngeneic recipients. These observations provide additional evidence that clonal deletion of autoaggressive T cell clones in the thymus is leaky. In this case allowing potentially autoaggressive T cell clones specific for S100 beta, a non-myelin autoantigen expressed in the nervous system, thymus and many peripheral tissues, to become an intrinsic component of the normal immune repertoire. PMID- 9199974 TI - Use of eluted peptide sequence data to identify the binding characteristics of peptides to the insulin-dependent diabetes susceptibility allele HLA-DQ8 (DQ 3.2). AB - HLA-DQ8 (A1*0301, B1*0302) and -DQ2 (A1*0501, B1*0201) are both associated with diseases such as insulin-dependent diabetes mellitus and coeliac disease. We used the technique of pool sequencing to look at the requirements of peptides binding to HLA-DQ8, and combined these data with naturally sequenced ligands and in vitro binding assays to describe a novel motif for HLA-DQ8. The motif, which has the same basic format as many HLA-DR molecules, consists of four or five anchor regions, in the positions from the N-terminus of the binding core of n, n + 3, n + 5/6 and n + 8, i.e. P1, P4, P6/7 and P9. P1 and P9 require negative or polar residues, with mainly aliphatic residues at P4 and P6/7. The features of the HLA DQ8 motif were then compared to a pool sequence of peptides eluted from HLA-DQ2. A consensus motif for the binding of a common peptide which may be involved in disease pathogenesis is described. Neither of the disease-associated alleles HLA DQ2 and -DQ8 have Asp at position 57 of the beta-chain. This Asp, if present, may form a salt bridge with an Arg at position 79 of the alpha-chain and so alter the binding specificity of P9. HLA-DQ2 and -DQ8 both appear to prefer negatively charged amino acids at P9. In contrast, HLA-DQ7 (A1*0301, B1*0301), which is not associated with diabetes, has Asp at beta 57, allowing positively charged amino acids at P9. This analysis of the sequence features of DQ-binding peptides suggests molecular characteristics which may be useful to predict epitopes involved in disease pathogenesis. PMID- 9199975 TI - Immunologic characterization and functional properties of murine antibodies raised against deleted mutants of human beta 2-glycoprotein I. AB - beta 2-Glycoprotein I (beta 2GPI) is a 50 kDa molecule proposed as a principal target of 'autoimmune' antiphospholipid antibodies (aPL). We have used deleted mutants (DM) representing different domains of beta 2GPI (I-IV, IV-V and V) for immunization of naive mice and studied the characteristics of the respective murine IgG preparations in comparison with affinity-purified IgG from two patients with primary antiphospholipid syndrome. Immunization with beta 2GPI and with the DM produced anti-beta 2GPI antibodies, part of which reacted with negatively charged phospholipids (PL), whereas reactivity with cardiolipin was evident only in the IgG from mice immunized with beta 2GPI. These results are consistent with the presumption that aPL are induced following the in vivo association of beta 2GPI (used for immunization) with resident negatively charged PL. Accordingly, DM which either lack the PL binding site or aPL attachment locus did not elicit, upon immunization, antibodies reactive with PL. Further, murine anti-beta 2GPI IgG and human 'autoimmune' aPL were similar, albeit not identical, in terms of DM requirement for PL binding and charge dependency. Murine antibodies and human aPL, regardless of their binding characteristics, were found to bind significantly to platelets upon their activation with thrombin and to promote platelet activation. The results of the current study emphasize the dissimilarities between human 'autoimmune' aPL and murine anti-beta 2GPI. Thus, anti-beta 2GPI antibodies to different DM as well as human aPL are capable of binding and activating human platelets provided beta 2GPI is present. PMID- 9199976 TI - Urticaria pigmentosa. PMID- 9199977 TI - Granuloma annulare. PMID- 9199978 TI - Methods for assessing erythema: a critique of parametric and nonparametric techniques. PMID- 9199979 TI - Nehushtan: the Tower of Babel, diversity or guidelines. PMID- 9199980 TI - Perforating granuloma annulare. AB - BACKGROUND: Perforating granuloma annulare (PGA) is considered an histologic subtype of granuloma annulare (GA) and it is described as a very rare disease, usually of children, affecting the dorsum of the hands. Mechanisms leading to perforation are unknown. Our experience suggested a different clinical presentation, so we decided to review our patients and the cases published. METHODS: We present six cases of PGA and review 52 PGA cases reported in the literature. Data regarding sex, age, time of evolution of disease, clinical features, laboratory findings, treatment, and follow-up were collected. The significance level was determined by the chi 2 or Student t test when appropriate. RESULTS: The prevalence of PGA could be up to 5% of GA. Pustular like lesions can be found in 26% of cases, and scars in 37%; papular, umbilicated, and crusted lesions being the most common finding. While PGA appears as a single lesion in only 9% of cases, and half of the patients are older than 30 years. In GA 50% of cases present as a single lesion and 80% of patients are younger than 30 years. CONCLUSIONS: PGA is different to GA not only histologically but also clinically. It is a disseminated disease, affecting both children and adults, which is characterized by the presence of multiple papules, most of them umbilicated and/or crusted, and characteristically pustular lesions and scars. Histology suggests that the superficial localization of the necrobiotic granuloma leads to the epidermal perforation. Treatment is disappointing. PMID- 9199981 TI - Leukocytoclastic vasculitis--correlation between different histologic stages and direct immunofluorescence results. AB - BACKGROUND: Leukocytoclastic vasculitis is a well-known clinico-pathologic entity. A good correlation between clinical and direct immunofluorescence (DIF) findings has been shown only in the early stages of vasculitis. Our purpose was to determine the correlation between different stages of vasculitis, etiology of vasculitis, and DIF findings. METHODS: Histologic and DIF studies were performed and evaluated from 40 patients with leukocytoclastic vasculitis. RESULTS: Thirty seven out of 40 patients (92%) showed positive DIF findings in the blood vessel walls. Eight patients were in the early stage of vasculitis and exhibited deposits mainly of fibrinogen, C3, and IgM. Seventeen patients were at the fully developed vasculitis stage and showed albumin, fibrinogen, and IgG deposits. Fifteen patients were in the late stage of vasculitis and showed deposits of mainly fibrinogen and C3 in the blood vessel walls. CONCLUSIONS: The present study demonstrates that DIF examination is a very sensitive test in the diagnosis of vasculitis, and can be used in all stages of vasculitis and not only in the early stages as has been shown in previous studies. PMID- 9199982 TI - Clinical characteristics of progressive vitiligo. AB - BACKGROUND: Vitiligo is an acquired hypopigmentary disorder with a progressive clinical course. Various clinical characteristics and their significance in the progression of vitiligo were evaluated. METHODS: A clinical study was carried out on 400 patients who had visited the Vitiligo Special Clinic of Severance Hospital, Yonsei University College of Medicine. Questionnaires were completed regarding progression, sex, family history, clinical type, onset age, duration of disease, Koebner's phenomenon, leukotrichia, and mucosal involvement, and the results were recorded. RESULTS: Progression of vitiligo was seen in 355 patients (88.8%); 45 patients (11.2%) did not show progression. No difference in sex, onset age, or leukotrichia was noted between the two groups; however, patients with positive family histories, nonsegmental clinical type, longer duration, Koebner's phenomenon, and mucous membrane involvement showed more progression of vitiligo. CONCLUSIONS: Clinical characteristics such as family history, clinical type, duration of disease, Koebner's phenomenon, and mucous membrane involvement, may be relevant in predicting the progression or prognosis of vitiligo. PMID- 9199983 TI - Neurocutaneous melanosis and Dandy-Walker syndrome in an infant. PMID- 9199984 TI - The Olmsted syndrome. PMID- 9199985 TI - Perforation of large and small bowel in Henoch-Schonlein purpura. PMID- 9199986 TI - Subcutaneous angiocentric T-cell lymphoma associated with fatal hemophagocytic syndrome. PMID- 9199987 TI - Plaque and postauricular nodular mucinosis associated with lupus erythematosus. PMID- 9199988 TI - Facial bullous systemic lupus erythematosus. PMID- 9199989 TI - Steatocystoma multiplex localized only in the face. PMID- 9199990 TI - Clothing as protection from ultraviolet radiation: which fabric is most effective? AB - BACKGROUND: With increasing levels of solar ultraviolet radiation (UVR) in the earth's near environment and evidence that exposure to UVR contributes to skin cancer, cataracts, and photoaging, protection of the skin is imperative during exposure to the sun. The purpose of this study was to examine the effectiveness of various fabrics in screening UVR and to determine if specific characteristics of fabric are directly related to the amount of protection provided. METHODS: Transmission of UVR was measured using spectrophotometric techniques. This transmission, as a function of wavelength over the range 250-400 nm, was weighted with solar and biological spectral data to determine a "sun protection factor" (SPF) for each fabric. RESULTS: The transmission of UVR through fabric depends on the wavelength and varies with factors such as type of fiber, fabric mass, cover, and color. CONCLUSIONS: Of 28 white fabrics tested, 19 offered less protection than a sunscreen with SPF 15. Polyester fabrics offered increased protection over cotton. The presence of dyes increased protection considerably. PMID- 9199992 TI - Laser treatment for further depigmentation in vitiligo. AB - BACKGROUND: In patients with vitiligo, sometimes the greatest part of the skin has already lost its melanocytes. The remaining pigmented patches can be removed by using strong bleaching creams, but many adverse events have been reported with this treatment. A new depigmentation therapy could be treatment with a Fluby laser. METHODS: Before treatment, the patients filled out a questionnaire about their vitiligo history. Eight patients with remaining pigmentation of the arms, hands, and face were treated once with a Ruby laser. Patients were monitored for developing repigmentation during the 9 months after treatment. RESULTS: In patients with a positive Koebner phenomenon, a permanent state of depigmentation was reached after laser therapy. None of the treated patients showed severe side effects. CONCLUSIONS: Ruby laser treatment can be an effective, fast, and safe method for removing cosmetically disturbing remnants of normal pigmentation in vitiligo patients with a positive Koebner phenomenon. PMID- 9199991 TI - Extracorporeal photochemotherapy in progressive systemic sclerosis: a follow-up study. AB - BACKGROUND: Extracorporeal photochemotherapy (photopheresis), an immune modulating therapy, has been demonstrated to elicit a therapeutic response in the treatment of several autoimmune disorders. We evaluated the use of photopheresis in the treatment of patients with progressive systemic sclerosis (PSS; scleroderma). METHODS: Five patients with early-onset, diffuse PSS were treated with photopheresis on 2 successive days monthly for an average of 59 months (range 54-89 months). We initially reported the response this group of patients had to photopheresis treatment at an average of 11 months (range 6-21 months). RESULTS: An improvement or stabilization was noted in most patients in skin thickening, joint mobility, pulmonary function studies, oral aperture, functional index, as well as symptoms including Raynaud's phenomenon, dyspnea, fatigue, dysphagia, arthralgias, and cutaneous ulcers. Renal function tests remained within normal range. A total of 296 monthly treatments were administered without significant toxicity. CONCLUSIONS: This study suggests that extended use of extracorporeal photochemotherapy in the management of early-onset, diffuse PSS is well tolerated and may provide an increasingly beneficial clinical outcome. PMID- 9199993 TI - Effective management of marked lymphedema of the leg. AB - BACKGROUND: Several conservative and surgical treatment methods have been demonstrated to be useful in the management of lymphedema. METHODS: In a patient with an enormous lymphedema of the leg, we first used complex decongestive physiotherapy as proposed by Foldi et al. in 1989, and then followed it with surgery. RESULTS: Satisfactory results were obtained in a short period of time and were maintained during the 2-year follow-up. CONCLUSIONS: We recommend a combined medical, surgical, and physiotherapeutic approach in patients with marked lymphedema. PMID- 9199994 TI - Complications of 585-nm pulsed dye laser therapy. AB - BACKGROUND: Port-wine stains (PWS) are a congenital, progressive ectasia of the superficial cutaneous vascular plexus that occur in 0.3%-0.5% of children at birth. They should be considered a disease with physical and psychologic complications. Treatment with the flashlamp pulsed dye laser (FPOL) has proven to be effective. Because treatment results in selective vascular injury, the risk of complications is minimal. The purpose of this study was to determine the cutaneous side-effects and complications and their frequency in patients with PWS treated with the FPDL. Eighty-nine patients who had been treated with the FPDL for PWS were included in this study. METHODS: Patients were treated at 3-month intervals using slightly overlapping pulses with fluences of 6.0-9.0 J/cm2. Treating physicians were responsible for evaluating the patients. All patients were reviewed and side-effects and complications were recorded. RESULTS: Pigmentary changes were the most frequently observed complications. Hyper- and hypopigmentation appeared in 16.8% and 2.4% of patients, respectively. Cutaneous depressions occurred in 2.2% of patients. One patient developed a hypertrophic scar, and another patient presented with transient cutaneous pustulosis. All these changes usually resolved spontaneously and completely in a few months. CONCLUSIONS: Most of the side-effects and complications produced by the FPDL are usually transitory, and it is essentially a very secure technique. PMID- 9199995 TI - Wound measurement. PMID- 9199996 TI - Dermatophytes in Poland. PMID- 9199997 TI - Risk factors for hip fracture in a Japanese cohort. AB - Risk factors for hip fracture were determined from a Japanese cohort. A cohort of 4573 people (mean age 58.5 +/- 12.2) who participated in the Adult Health Study in 1978-1980 were subsequently followed by biennial examinations up to 1992. Fifty-five incident hip fractures not due to traffic accidents were identified by medical records during the follow-up period. Poisson regression analysis showed that baseline low body mass index (BMI), regular alcohol intake, prevalent vertebral fracture, and having five or more children significantly increased the risk of hip fracture, and low milk intake and later age at menarche were marginally associated with increased fracture risk, after multivariable adjustment. Regular alcohol intake doubled the risk of hip fracture (relative risk 1.91, 95% confidence interval 1.07-3.42). Those individuals who had a vertebral fracture had 2.6 times higher risk than those who did not. The risk was 2.5 times higher among women who had five or more children than women with one or two. Body height, health status, marital status, intake of fish, coffee, tea, Japanese tea, smoking, exposure to atomic bomb radiation, and age at menopause were not associated with hip fracture. Relative risk for hip fracture decreased with decreasing number of preventable risk factors (low BMI, low milk intake, and regular alcohol intake). We conclude that many factors, such as BMI, milk intake, alcohol intake, prevalent vertebral fracture, age at menarche, and number of children, are related to the risk of hip fracture, and prevention programs need to focus on reducing preventable risk factors. PMID- 9199998 TI - Fractures among the elderly: an old problem. PMID- 9199999 TI - Cloning and sequencing of human PEX from a bone cDNA library: evidence for its developmental stage-specific regulation in osteoblasts. AB - Inactivating mutations of the neutral endopeptidase, PEX, have been identified as the cause of X-linked hypophosphatemia (XLH). Though the function of PEX is unknown, current information suggests that impaired renal phosphate conservation in XLH is due to the failure of PEX to either degrade an undefined phosphaturic factor or activate a novel phosphate-conserving hormone. The physiologically relevant target tissue for the XLH mutation has not been identified. An apparent intrinsic defect of osteoblast function in XLH implicates bone as a possible site of PEX expression. In the current investigation, we employed a polymerase chain reaction (PCR) strategy to amplify a PEX cDNA from a human bone cell cDNA library. We found that the human PEX cDNA encodes a 749 amino acid protein belonging to the type II integral membrane zinc-dependent endopeptidase family. The predicted PEX amino acid sequence shares 96.0% identify to the recently cloned mouse Pex cDNA and has 27-38% identity to other members of the metalloendopeptidase family. Using reverse transcriptase (RT)-PCR with PEX specific primers, we detected PEX transcripts in both human osteosarcoma-derived MG-63 osteoblasts and in differentiated mouse MC3T3-E1 clonal osteoblasts but not in immature MC3T3-E1 preosteoblasts. The association of impaired mineralization of bone in XLH and the apparent developmental stage-specific expression of PEX in osteoblasts suggest that bone is a physiologically relevant site of PEX expression and that PEX may play an active role in osteoblast-mediated mineralization. PMID- 9200000 TI - Efferent targets of osseous CGRP-immunoreactive nerve fiber before and after bone destruction in adjuvant arthritic rat: an ultramorphological study on their terminal-target relations. AB - We report the ultramorphological characterization of the terminal-target relation of sensory peptidergic nerve fibers in healthy and diseased osseous tissues. Bone tissue sections were immunoelectronmicroscopically investigated for calcitonin gene-related peptide (CGRP), a neuropeptide widely distributed in sensory peptidergic fibers. Ultramorphological relation of the osseous CGRP immunoreactive (ir) nerve terminals and their target cells was comparatively analyzed using healthy, arthritic, and postarthritic bone specimens from control and adjuvant-induced arthritic rats. Terminal-like profiles of the osseous CGRP ir axons were evidenced in direct contact with the metaphyseal osteoblasts and osteoclasts of the control animals. Terminal-like profiles were also noted in the vicinity of the periosteal lining cells. Nonterminal-like profiles did not make intimate spatial relation to the cells/structures surrounding the nerve. Osseous CGRP-ir terminals and axons, which are either uncovered or thinly ensheathed by the supportive tissues, were extensively degenerated in adjuvant-induced infiltration, whereas larger fibers were relatively resistant. Numerous CGRP-ir axons with distinctive features reinnervated the postarthritic, ossifying periosteum. CGRP-ir axons appeared to reinnervate the eroded surface of metaphyseal bone and cartilage as early as the recruited osteoblasts resume osteogenesis in the postarthritic metaphysis. The observed terminal-target relations in the healthy and diseased bone tissues give an ultramorphological basis for the putative trophic, modulatory actions of CGRP innervation of the bone cells. PMID- 9200001 TI - Measurement of bone degradation products in serum using antibodies reactive with an isomerized form of an 8 amino acid sequence of the C-telopeptide of type I collagen. AB - An enzyme-linked immunosorbent assay for measuring type I collagen degradation products in serum (S-ELISA) was developed. The assay uses a high affinity polyclonal antibody which reacts with an isomerized form of an 8 amino acid sequence of the C-telopeptides of type I collagen (EKAHD-beta-GGR). Cross reactivity to a nonisomerized synthetic peptide form of the 8 amino acid sequence is less than 0.2%. Values obtained in a group of premenopausal women (age, 33.3 +/- 3.11 years) were 69 +/- 24 ng/ml(n = 22). In a group of early postmenopausal women (age, 51.8 +/- 1.88 years) values obtained were 125 +/- 43 ng/ml (n = 46), which represents an increase of 81% (p < 0.001). Values found in untreated patients with Paget's disease were 234 +/- 95 ng/ml (n = 15), and for primary hyperparathyroidism we found 335 +/- 82 ng/ml (n = 10). Intravenous administration of a bisphosphonate (Pamidronate) to Paget's disease patients for 3 days was reflected in the S-ELISA by a decrease in the values of 55% when compared with values before treatment (n = 15). Following treatment with another bisphosphonate (Alendronate) for 6 months, values were decreased to 48 +/- 19 ng/ml (n = 12), which corresponds to a 62% decrease. Clinical results presented in this context support that the assay is a sensitive and specific index of bone resorption. It may, therefore, prove useful in the follow up of treatment of patients with metabolic bone diseases and in the clinical investigation of osteoporosis. PMID- 9200002 TI - Plasmin-mediated proteolysis of osteocalcin. AB - Plasmin cleaves osteocalcin at a site within its carboxyl end, thus creating an N midterminal 1-43 and a short C-terminal 44-49 peptides. The products of the cleavage were identified by matrix assisted laser desorption ionization time of flight mass spectrophotometry and by reversed phase high performance liquid chromatography followed by N-terminal sequence determination. When separated by sodium dodecyl sulfide-polyacrylamide gel electrophoresis in the presence of reducing agents, large (LF; N-midterminal) and a small molecular weight (SF; C terminal) fragments can be identified. The major cleavage site involves arg43 arg44 amino acid residues, and the resulting 44-49 C-terminal fragment appears as a slow migrating band on native gels (SFnat). Elevated levels of calcium ion inhibit the plasmin-mediated lysis of osteocalcin. Plasmin-mediated cleavage of osteocalcin occurs both in solution and when bound to hydroxyapatite. Both osteocalcin cleavage products detach from the hydroxyapatite substrate. Diisopropyl fluorophosphate-inhibited plasmin does not displace osteocalcin from the hydroxyapatite surface. Previously, the C-terminal pentapeptide has been shown to be chemotactic for bone cells while bone particles lacking osteocalcin were resistant to bone resorption. We therefore hypothesize that the plasmin mediated digestion of free and hydroxyapatite-bound osteocalcin could play a role in the regulation of bone remodeling. PMID- 9200003 TI - The vitamin D receptor start codon polymorphism (FokI) and bone mineral density in premenopausal American black and white women. AB - This study examines the association between bone mineral density (BMD) and a start codon polymorphism (SCP) at the translation initiation site of the vitamin D receptor (VDR) gene. The thymine/cytosine (T/C) polymorphism in the first of two start (ATG) codons can be detected by a restriction fragment length polymorphism (RFLP) using the endonuclease FokI, which recognizes ATG as part of its restriction site. F indicates absence of the first ATG and a VDR that is shorter by three amino acids. The FokI genotype was determined in 154 premenopausal American women (72 black and 82 white) who were 20-40 years old. BMD of the total body, femoral neck, and lumbar spine were measured by dual energy X-ray absorptiometry. The distribution of the SCP genotypes differed significantly by race (p < 0.001): 4% of blacks versus 18% of whites were ff homozygous and 65% of blacks versus 37% of whites were FF homozygous. There was no statistically significant interaction between race and SCP genotype in analyses of BMD at any skeletal site. In the group as a whole, the ff women had femoral neck BMD that was 7.4% lower than that of the FF women. The ff white women had total body BMD values that were 4.3% lower and femoral neck values that were 12.1% lower than FF white women. Total body and femoral neck BMD did not differ significantly by genotype in black women, and spine BMD did not differ by genotype in either race. Addition of the SCP genotype to analysis of covariance models comparing BMD of the black and white women reduced estimated differences in femoral neck BMD between the two groups by about 35%. In conclusion, the SCP polymorphism, detected with the endonuclease FokI, appears to influence peak bone density, particularly at the femoral neck. Racial differences in its distribution may explain some of the racial difference in femoral neck BMD. PMID- 9200004 TI - The BsmI vitamin D receptor restriction fragment length polymorphism (bb) influences the effect of calcium intake on bone mineral density. AB - Previous studies of the vitamin D receptor (VDR) polymorphisms and bone mineral density (BMD) have suggested that there may be differences in calcium absorption among groups of women with different VDR genotypes, and that the association may be stronger in younger women. To investigate the association between the VDR polymorphisms and BMD, this study was undertaken in the Framingham Study Cohort and a group of younger volunteers. Subjects from the Framingham Study (ages 69-90 years) included those who underwent BMD testing and who had genotyping for the VDR alleles (n = 328) using polymerase chain reaction methods and restriction fragment length polymorphisms with BsmI (B absence, b presence of cut site). A group of younger volunteer subjects (ages 18-68) also underwent BMD testing and VDR genotyping (n = 94). In Framingham Cohort subjects with the bb genotype, but not the Bb or BB genotypes, there were significant associations between calcium intake and BMD at five of six skeletal sites, such that BMD was 7-12% higher in those with dietary calcium intakes greater than 800 mg/day compared with those with intakes < 500 mg/day. The data also suggested that BMD was higher in persons with the bb genotype only in the group with calcium intakes above 800 mg/day. No significant differences were found in the Framingham Cohort for age-, sex-, and weight-adjusted BMD at any skeletal site between those with the BB genotype and those with the bb genotype regardless of 25-hydroxyvitamin D levels or country of origin. In the younger volunteers, BMD of the femoral neck was 5.4% higher (p < 0.05) in the bb genotype group compared with the BB group and 11% higher (p < 0.05) in males with the bb genotype compared with the BB group. There were no significant differences at the lumbar spine. In this study, the association between calcium intake and BMD appeared to be dependent upon VDR genotype. The findings of an association between dietary calcium intake and BMD only in the bb genotype group suggests that the VDR genotype may play a role in the absorption of dietary calcium. Studies that do not consider calcium intake may not detect associations between VDR genotype and BMD. In addition, the association between VDR alleles and BMD may become less evident in older subjects. PMID- 9200005 TI - Inhibition of bone resorption by pamidronate cannot restore normal gain in cortical bone mass and strength in tail-suspended rapidly growing rats. AB - To clarify how the changes in bone formation and resorption affect bone volume and strength after mechanical unloading, the effect of inhibition of bone resorption by a potent bisphosphonate, pamidronate, on bone mineral density (BMD), histology, and strength of hind limb bones was examined using tail suspended growing rats. Tail suspension for 14 days reduced the gain in the BMD of the femur at both the metaphysis rich in trabecular bone and the diaphysis rich in cortical bone. Treatment with pamidronate increased the total BMD as well as that of the metaphysis of the femur but had almost no effect on the BMD of the diaphysis in both control and tail-suspended rats. Histological examinations revealed that 14-day tail suspension caused a loss of secondary cancellous bone with a reduction in the trabecular number and thickness in comparison with control rats. In the femoral diaphysis, the diameter and cortical bone thickness increased to a lesser degree in tail-suspended rats when compared with rats without tail suspension, and a marked reduction in bone formation and the layers of alkaline phosphatase-positive cells was observed at the periosteal side. Pamidronate treatment increased secondary cancellous bone but could not restore normal growth-induced periosteal bone apposition and bone strength. Because the material strength of the femoral diaphysis at the tissue level was not affected by pamidronate treatment, the inability of pamidronate to prevent the reduction in physical strength of the femoral diaphysis does not appear to be due to a change in the quality of newly formed bone. These results demonstrate that tail suspension reduces the growth-induced periosteal modelling drift and that the antiresorptive agent pamidronate is unable to restore normal periosteal bone apposition. PMID- 9200006 TI - Regional responsiveness of the tibia to intermittent administration of parathyroid hormone as affected by skeletal unloading. AB - To determine whether the acute inhibition of bone formation and deficit in bone mineral induced by skeletal unloading can be prevented, we studied the effects of intermittent parathyroid hormone (PTH) administration (8 micrograms/100 g/day) on growing rats submitted to 8 days of skeletal unloading. Loss of weight bearing decreased periosteal bone formation by 34 and 51% at the tibiofibular junction and tibial midshaft, respectively, and reduced the normal gain in tibial mass by 35%. Treatment with PTH of normally loaded and unloaded animals increased mRNA for osteocalcin (+58 and +148%, respectively), cancellous bone volume in the proximal tibia (+41 and +42%, respectively), and bone formation at the tibiofibular junction (+27 and +27%, respectively). Formation was also stimulated at the midshaft in unloaded (+47%, p < 0.05), but not loaded animals (-3%, NS). Although cancellous bone volume was preserved in PTH-treated, unloaded animals, PTH did not restore periosteal bone formation to normal nor prevent the deficit in overall tibial mass induced by unloading. We conclude that the effects of PTH on bone formation are region specific and load dependent. PTH can prevent the decrease in cancellous bone volume and reduce the decrement in cortical bone formation induced by loss of weight bearing. PMID- 9200007 TI - Calcaneal bone mineral density predicts fracture occurrence: a five-year follow up study in elderly people. AB - A 5-year follow-up study investigated calcaneal bone mineral density (BMD) and changes in BMD in relation to fracture occurrence. The subjects comprised two cohorts born in 1914 and 1910 living in the city of Jyvaskyla in central Finland. One hundred and three men (82%) and 188 women (73%), aged 75, and 57 men (74%) and 136 women (65%), aged 80, of the eligible population participated in the baseline bone measurements. The follow-up bone measurements were obtained for 59 men (68%) and 119 women (66%), aged 80 years, and for 21 men (53%) and 61 women (48%), aged 85 years. During the follow-up period, 8 men and 36 women from the younger and 11 men and 24 women from the older cohort sustained at least one fracture. When the baseline levels of BMD were related to fracture occurrence, the results clearly showed that with increased BMD values the probability of fracture decreased. Where men and women had similar BMD values, they also had a similar fracture probability. Except for one woman in the older cohort, none of those who had initial BMD values more than 1 standard deviation above the mean for their age developed a fracture during the follow-up period. The mean annual decrease in BMD was greater in the women (2.5-2.7%) than in the men (0.8-1.0%). The BMD change tended to associate with fracture occurrence only in the 75-year old women (p = 0.075). The results suggest that calcaneus BMD can be used as a predictor of fracture occurrence in 75- to 80-year-old men and women. However, associating fractures with the change in BMD was difficult due to the limited number of survivors and initial differences in BMD values. PMID- 9200008 TI - Relationship of bone turnover to bone density and fractures. AB - To assess the influence of bone turnover on bone density and fracture risk, we measured serum levels of osteocalcin (OC), bone alkaline phosphatase (BAP), and carboxy-terminal propeptide of type I procollagen (PICP), as well as 24-h urine levels of cross-linked N-telopeptides of type I collagen (NTx) and the free pyridinium cross-links, pyridinoline (Pyd) and deoxypyridinoline (Dpd), among 351 subjects recruited from an age-stratified random sample of Rochester, Minnesota women, PICP, NTx, and Dpd were negatively associated with age among the 138 premenopausal women. All of the biochemical markers were positively associated with age among the 213 postmenopausal women, and the prevalence of elevated turnover (> 1 standard deviation [SD] above the premenopausal mean) varied from 9% (PICP) to 42% (Pyd). After adjusting for age, most of the markers were negatively correlated with bone mineral density (BMD) of the hip, spine, or forearm as measured by dual-energy X-ray absorptiometry, and women with osteoporosis were more likely to have high bone turnover. A history of osteoporotic fractures of the hip, spine, or distal forearm was associated with reduced hip BMD and with elevated Pyd. After adjusting for lower BMD and increased bone resorption, reduced bone formation as assessed by OC was also associated with prior osteoporotic fractures. These data indicate that a substantial subset of elderly women has elevated bone turnover, which appears to adversely influence BMD and fracture risk. Combined biochemical and BMD screening may provide better prediction of future fracture risk than BMD alone. PMID- 9200009 TI - Fractures after thyroidectomy in men: a population-based cohort study. AB - Bone mass is purportedly reduced by an endogenous or exogenous excess of thyroid hormone or, perhaps, by calcitonin deficiency. Patients who have undergone thyroidectomy could be subject to all of these effects, yet their practical implications in terms of fracture risk are poorly defined. Interpretation is further hampered by the focus on women, where results may be influenced by involutional osteoporosis. Consequently, we assessed the potential for fractures among the 136 Rochester, Minnesota men who underwent thyroidectomy between 1935 and 1979, relative to a group of age-matched control men from the community. With 2194 person-years of follow-up in each group, survival free of any fracture of vertebra, proximal humerus, distal forearm, pelvis, or proximal femur was similar in the two groups (p = 0.23), and the relative risk of any of these fractures for thyroidectomized patients versus their controls was increased only 1.5-fold (95% CI, 0.7-3.2). The difference was entirely accounted for by a statistically significant excess of proximal femur fractures in the men with thyroidectomy. Risk factors for fractures among men with thyroidectomy included greater age at surgery, greater extent of surgery, and the presence of risk factors for secondary osteoporosis. Thus, thyroidectomy, performed mainly for adenoma or goiter, seems to have little overall influence on the risk of age-related fractures in men. However, the association with hip fractures requires further evaluation. PMID- 9200010 TI - Attitudes and beliefs of family physicians and gynecologists in relation to the prevention and treatment of osteoporosis. AB - The objective of this study was to evaluate the attitudes and beliefs of primary care physicians (PCPs) and obstetricians/gynecologists (O&Gs) in relation to the prevention and treatment of osteoporosis (OP) in postmenopausal women. A survey was mailed to a random sample of PCPs and to all O&Gs registered in the province of Alberta (Canada). The survey evaluated their practice patterns using closed ended questions, Likert scaled items, and two case studies. Cases 1 and 2 were 52 year-old and 62-year-old healthy postmenopausal women, respectively, with no known risks for OP. Neither had received hormone replacement therapy (HRT). One hundred fifty-seven PCPs and 57 O&Gs participated in the study. Thirty-eight percent of the PCPs and 32% of the O&Gs stated that they never requested bone mineral density measurements (BMDm) in early postmenopausal women. Most would request BMDm only in the presence of risk factors. The most important criteria to request BMDm were chronic glucocorticoid use and recent fractures. For case 1, 7% of the PCPs and 11% of the O&Gs would request BMDm; 76% of the PCPs and 80% of the O&Gs would recommend HRT. For case 2, 29% of the PCPs and 47% of the O&Gs would request BMDm (p = 0.01); 43% of the PCPs and 49% of the O&Gs would prescribe HRT. In general, O&Gs were more inclined to intervene in relation to BMDm and HRT. O&Gs were also more likely to be influenced by clinical trials than PCPs (p < 0.001). Our findings show variations in the patterns of practice of physicians in relation to the prevention of OP. In general, use of densitometry appears to be low. The results of the case studies suggest that individual physician perceptions may be more influential than patient characteristics when requesting BMDm and prescribing HRT, particularly in older postmenopausal women. This group of healthy older women have approximately equal odds of being offered versus not being offered BMDm and HRT according to the physician they consult. PMID- 9200011 TI - Expression of nitric oxide synthase isoforms in bone and bone cell cultures. AB - Recent work has shown that nitric oxide (NO) acts as an important mediator of the effects of proinflammatory cytokines and mechanical strain in bone. Although several bone-derived cells have been shown to produce NO in vitro, less is known about the isoforms of NO synthase (NOS), which are expressed in bone or their cellular distribution. Here we investigated the expression, cellular localization, and regulation of NOS mRNA and protein in cultured bone-derived cells and in bone tissue sections. We failed to detect inducible NOS (iNOS) protein in normal bone using immunohistochemical techniques, even though low levels of iNOS mRNA were detected by sensitive reverse transcribed polymerase chain reaction (RT-PCR) assays in RNA extracted from whole bone samples. Cytokine stimulation of bone-derived cells and bone explant cultures caused dramatic induction of iNOS mRNA and protein in osteoblasts and bone marrow macrophages, but no evidence of iNOS expression was seen in osteoclasts by immunohistochemistry or in situ hybridization. Endothelial NOS (ecNOS) mRNA was also detected by RT-PCR in whole bone, and immunohistochemical studies showed widespread ecNOS expression in bone marrow cells and trabecular lining cells in vivo. Related studies in vitro confirmed that ecNOS was expressed in cultured osteoblasts, stromal cells, and osteoclasts. Neuronal NOS mRNA was detected by RT PCR in whole bone, but we were unable to detect nNOS protein in bone cells in vivo or in studies of cultured bone-derived cells in vitro. In summary, our data show that mRNAs for all three NOS isoforms are expressed in bone and provide evidence for differential expression and regulation of the enzymes in different cell types. These findings confirm the likely importance of the L-arginine-NO pathway as a physiological mediator of bone cell function and demonstrate that it may be possible to exert differential effects on osteoblast and osteoclast activity in vivo by differential targeting of constitutive and inducible NOS isoforms by selective NOS inhibitors. PMID- 9200012 TI - Specific inhibitors of vacuolar H(+)-ATPase trigger apoptotic cell death of osteoclasts. AB - Osteoclasts are multinucleated bone-resorbing cells that play a critical role in bone remodeling. Specific inhibitors of vacuolar H(+)-ATPase (V-ATPase), concanamycin A and bafilomycin A1, abolish bone resorption by osteoclasts. In this study, we examined whether these V-ATPase inhibitors trigger apoptotic cell death in osteoclasts, using murine osteoclast-like multinucleated cells (OCLs) formed in vitro. Acridine orange staining revealed that the treatment of OCLs with concanamycin A resulted in chromatin condensation and alterations in nuclear morphology within a few hours. The TdT-mediated dUTP-nick-end labeling (TUNEL) reaction confirmed the apoptotic features of OCLs treated with concanamycin A. The accelerated apoptotic cell death induced by concanamycin A occurred in OCLs treated with interleukin-1 alpha or macrophage colony-stimulating factor as well, which are known to elongate the survival time of osteoclasts. In contrast, these inhibitors did not induce cell death of osteoblastic cells isolated from mouse calvaria. These results suggest that functional impairment of V-ATPase triggers apoptotic cell death in osteoclasts. PMID- 9200013 TI - Involvement of alpha5beta1 integrin in matrix interactions and proliferation of chondrocytes. AB - Integrins are cell surface receptors involved in cellular processes including adhesion, migration, and matrix assembly. In the present study, we analyzed the possible involvement of alpha 5 beta 1 integrin in the regulation of chondrocyte adhesion, spreading, and proliferation. We found that rabbit growth plate chondrocytes were able to attach to substrates coated with type I collagen, type II collagen, or fibronectin within 24 h of culture. During this time period, attachment to fibronectin appeared to be dependent on alpha 5 beta 1 integrin, whereas adhesion to collagens was not. By day 3 of culture, chondrocytes spread onto all the substrates tested. We found that regardless of the nature of the substrate, cell spreading was reversed by treatment with RGD peptide or antibodies against alpha 5 beta 1 or fibronectin, indicating that cell spreading involved alpha 5 beta 1 and fibronectin endogenously produced and deposited by the chondrocytes themselves. Colony formation by chondrocytes in soft agar was inhibited by treatment with RGD peptides or BIIG2, an antibody that interferes with alpha 5 beta 1 integrin-ligand interactions. Furthermore, DNA content was decreased by treatment with anti-fibronectin antibody in micromass culture of chondrocytes. Immunohistochemical analysis on tissue sections revealed that the alpha 5 subunit was particularly abundant in the proliferative and hypertrophic zones of growth plate. The results of the study indicate that alpha 5 beta 1 integrin plays multiple roles in chondrocyte behavior and function and appears to be involved in the regulation of both chondrocyte-matrix interactions and proliferation. PMID- 9200014 TI - Electrophysiological responses of human bone cells to mechanical stimulation: evidence for specific integrin function in mechanotransduction. AB - Bone cells respond to mechanical stimuli, but the transduction mechanisms responsible are not fully understood. Integrins, a family of heterodimeric transmembrane glycoproteins, which link components of the extracellular matrix with the actin cytoskeleton, have been implicated as mechanoreceptors. We have assessed the roles of integrins in the transduction of cyclical mechanical stimuli to human bone cells (HBCs), which results in changes in membrane potential. HBC showed membrane depolarization following 0.104 Hz mechanical stimulation and membrane hyperpolarization following stimulation at 0.33 Hz. The membrane depolarization response involved tetrodotoxin-sensitive sodium channels and could be inhibited by antibodies against alpha V, beta 1, and beta 5 integrins. In contrast, the hyperpolarization response was inhibited by gadolinium and antibodies to the integrin-associated protein (CD47), alpha 5 and beta 1 integrin. Both responses could be abrogated by ARg-Gly-Asp (RGD) containing peptides, inhibition of tyrosine kinase activity, and disruption of the cytoskeleton. These results demonstrate differential electrophysiological responses of HBC to different frequencies of mechanical strain. Furthermore, they suggest that integrins act as HBC mechanoreceptors with distinct signaling pathways being activated by different frequencies of mechanical stimuli. PMID- 9200015 TI - Assessing cardiovascular status in the older adult with cognitive impairments. AB - When caring for older adults with cognitive impairments, nurses may find that their usual approaches to cardiovascular assessment are not effective. This article summarizes the geropsychiatric nursing literature about interventions that assist people with chronic cognitive impairments to maintain functional skills and decrease disruptive behavior patterns. A case study illustrates application of theory from behavioral gerontology and research from studies of persons with dementia to develop an approach to cardiovascular assessment that can assist clinicians in obtaining accurate data about cardiovascular status. PMID- 9200016 TI - Tissue oxygenation and routine nursing procedures in critically ill patients. AB - Maintaining adequate oxygenation to promote vital organ functions represents a common challenge for the critical care nurse. Critically ill patients with impaired cardiac function may be particularly vulnerable to tissue oxygen deprivation because they have limited ability to increase oxygen delivery when oxygen demands increase. Consequently, routine nursing procedures that increase oxygen requirements may have adverse effects on tissue oxygenation. Interventions that enhance patient tolerance to nursing procedures by supporting the balance between oxygen supply and demand promote physiologic adaptation and may prevent complications associated with hypoxia such as cardiac dysrhythmias, hypotension, and cardiac arrest. This discussion will focus on the principles of tissue oxygenation, the effects of nursing interventions on oxygen demand, and interventions that may enhance patient tolerance to routine nursing interventions. PMID- 9200017 TI - A multicultural perspective of cardiovascular disease. AB - Cardiovascular disease constitutes an emerging public health problem in most countries, even though morbidity and mortality profiles vary according to inherent risk factors. Urbanization and adoption of Western life style appear to predispose populations of all countries to greater risks for coronary heart disease and related conditions. The cultural composition of the United States is rapidly changing due to immigration. This growing ethnic diversity has placed new demands on the health care system for providing culturally sensitive care. In the United States, no ethnic group appears to be immune to cardiovascular disease. Cardiovascular risk factors of major American ethnic populations are discussed, and cultural beliefs that stress implications for nursing care of multicultural patients are introduced. PMID- 9200018 TI - Hypertension in an inner-city minority population. AB - This study describes an inner-city elderly minority population's awareness of hypertension and health perception compared with that of a younger hypertensive population. The elderly African American sample had a mean age of 71.6 years and systolic blood pressure of m = 143 mm Hg SD = 21. The younger sample averaged lower systolic readings (m = 129 mm Hg SD = 20). Overall health perception of both groups was positive (64%). Over 60% of the older people had been told that they were hypertensive. Because of the high percentage of aware hypertensive subjects on treatment, interventions focused on treatment adherence. Using the diffusion model, community awareness and strategies to remind people about their blood pressure were the focal interventions. PMID- 9200019 TI - Cardiovascular risk factors in homeless adults. AB - Homelessness is a growing problem in the United States, with population numbers ranging from 300,000 to several million. This article reviews five studies that focused on identification and assessment of cardiovascular risk factors in the homeless population. A case study presents the successful outcome of a homeless male with several risk factors. Assessment and intervention by a nurse practitioner led to his re-entry into the domiciled population. Implications for clinical practice and recommendations for further research are discussed. PMID- 9200020 TI - Considerations related to disability and exercise in elderly women with congestive heart failure. AB - Congestive heart failure is one of the most common diagnoses in older women. This article reviews physiologic and pathophysiologic factors that contribute to disability in older women with normal left ejection fraction congestive heart failure; the possibility that aerobic exercise training may be an effective means to reduce the disability experienced by these women is examined. Most literature has dealt with low-output ejection fraction congestive heart failure. Comparisons between low-output ejection fraction congestive heart failure and normal left ejection fraction congestive heart failure can help to clarify beneficial interventions. A physiologic model is proposed that includes peripheral and cardiac factors caused by heart disease, and factors caused by aging, that may account for increasing the disabling consequences and reducing exercise tolerance of older women with normal left ejection fraction congestive heart failure. The potential impact of exercise training on these factors is discussed. Directions for practice and further research are included. PMID- 9200021 TI - Factors contributing to rehospitalization of elderly patients with heart failure. AB - This article describes factors contributing to rehospitalizations of elderly patients with heart failure. Advanced practice nurses' logs, study questionnaires, and medical record summaries from a recent clinical trial provided rich, descriptive information about a variety of social and behavioral factors surrounding rehospitalization in these medically fragile older people. Medication and dietary nonadherence were factors affecting symptom appearance and rehospitalization. Social and behavioral factors, such as the absence of strong social support or motivation, contributed to nonadherence. These results suggest that social and behavioral factors must be identified and addressed in an individualized manner to prevent recurrent hospitalizations for elderly patients with heart failure. PMID- 9200022 TI - Blood pressure reactivity in urban youth during angry and normal talking. AB - Distinctions between male and female adult patterns of cardiovascular disease are well established. Although risk for cardiovascular disease begins early in life, little research has been focused on the beginning of gender difference patterns. This pilot study examined blood pressure reactivity, gender, and biologic maturity in adolescents talking about angry life circumstances and the usual progression of their day. Systolic and diastolic blood pressure increased during talking, and the addition of biologic maturity to the analysis distinguished males from females. Males had higher systolic blood pressures than did females throughout the protocol, and, generally, mature subjects had higher systolic and diastolic blood pressures than did less mature subjects. These pilot data confirm the effects of talking on blood pressure. The data suggest that biologic maturity enhances understanding of gender differences in adolescent blood pressure reactivity and continued study of the gender-blood pressure reactivity distinctions in adolescents may further understanding of adult patterns of gender differences in cardiovascular disease. PMID- 9200023 TI - Escherichia coli O157: lessons from the past 15 years. PMID- 9200024 TI - HIV genetic variation: clinical importance. PMID- 9200025 TI - HIV genetic variation: life at the edge. PMID- 9200026 TI - Antibody responses to different strains of coxsackie B4 virus in patients with newly diagnosed type I diabetes mellitus or aseptic meningitis. AB - Considerable differences in antibody responses measured by capture-IgM RIA and neutralization tests (NT) were seen in children with newly diagnosed type I (insulin-dependent) diabetes mellitus (IDDM) when five different strains of Coxsackie B4 virus (CBV-4) were used. The IgM positivity of the 160 patients varied between 3.7 and 10.0% (P < 0.05) and with the use of all strains, IgM was found in 13.2%. Matched controls showed a significantly lower frequency (P < 0.05), but there were no apparent differences between the strains, although no IgM was found against two strains. In the NT different results were also obtained in the IDDM children with use of the five strains. However, these results differed considerably from those of the RIA, indicating that different epitopes on the virus were involved. Serum specimens from 75 patients with aseptic meningitis from whom enteroviruses had been isolated, also showed varying IgM frequencies, ranging from 13.3 to 18.7%, but the differences between the strains were different to those found in the IDDM patients. We speculate that only certain strains of a serotype of CBV may be used to distinguish IDDM pathogenesis from that of other diseases. PMID- 9200027 TI - A comparison of acridine orange, wet microscopy and Gram staining in the diagnosis of bacterial vaginosis. PMID- 9200028 TI - Broviac catheter-related bacteraemias due to unusual pathogens in children with cancer: case reports with literature review. AB - Among 102 episodes of intravenous catheter related bacteraemias documented between January 1989 and July 1996 in children receiving antineoplastic chemotherapy or bone marrow transplantation at G. Gaslini Children's Hospital, Genoa, Italy, were identified seven episodes due to unusual pathogens: Bacillus circulans, Bacillus licheniformis, Brevibacterium casei, Flavimonas oryzihabitans, Porphyromonas asaccharolytica, Comamonas acidovorans and Agrobacterium radiobacter. Susceptibility to different antibiotics of all strains are reported. In all cases catheter removal was required for culture negativization. All episodes were diagnosed in absence of granulocytopenia. PMID- 9200029 TI - Haemophagocytic syndrome in patients infected with the human immunodeficiency virus: nine cases and a review. AB - We report nine cases of haemophagocytic syndrome (HS) in patients with human immunodeficiency virus infection, and review the 17 cases described to date in the literature. In 21 of 26 cases, HS developed during an advanced stage of immunodeficiency. Clinical and haematological signs are not specific in this setting, and the diagnosis relies on histological features, mainly bone marrow examination. An opportunistic infection was associated in three cases and a lymphoid malignancy in two of our nine cases. The role of the human immunodeficiency virus in the occurrence of the HS remains to be defined. The overall prognosis is poor as six of our nine patients died within 12 months of presentation. PMID- 9200030 TI - Acute meningococcal meningitis: analysis of features of the disease according to the age of 255 patients. Copenhagen Meningitis Study Group. AB - Clinical and laboratory features of acute meningococcal meningitis according to age were studied in 255 patients. Whereas males accounted for three out of five patients aged 0-4 years, females accounted for three out of four patients older than 50 years of age. All patients had clinical signs of nuchal rigidity and fever. Patients older than 30 years of age had less frequent petechiae (62%) than younger patients (81%). Furthermore, elderly patients above 50 years of age were prone to an obtunded mental state and a prolonged disease course with fever. Without relation to age, 2/3 had purulent meningitis and 2/3 had marked peripheral leucocytosis (> 15 x 10(9) cells/l); 90% of patients had at least one of these findings. The cellular inflammatory response in peripheral blood indicated a bacterial aetiology in > 95% of the cases. More than 80% of children and adults had abnormal CSF biochemical findings, but the level of protein and the glucose ratio (CSF/serum) were positively and negatively correlated to increasing age of the patient, respectively: thus, in children these biochemical markers may be unreliable in the differentiation between a bacterial and non bacterial aetiology. Thrombocytopenia (< 100.000 x 10(9)/I) was not associated with age, though the lowest platelet count was found in elderly patients. The case fatality rate was 7.5%, but neither age, sex nor sign of septicaemia was associated with fatality. Thrombocytopenia, a lowered coagulation index (< 0.5, factors II, VII, X), a moderate anaemia (haemoglobin < 11 g/dl), an obtunded mental state and a history of convulsions were poor prognostic factors; only anaemia was independently correlated to fatality so this should be considered as an important prognostic marker in the acute phase of meningococcal meningitis. PMID- 9200031 TI - Typhoid fever in hospitalized children in Singapore. AB - A study was conducted to determine the clinical features and severity of typhoid fever in children admitted to the Communicable Disease Centre, Singapore. Over a 5 year period (1990-94), 40 children had documented culture-proven typhoid fever. Nine of the 40 children (22.5%) were below the age of 5 years. The predominant presenting symptoms were fever and diarrhoea. Pneumonia and ileus were the only complications observed. No patient in the series died. Ten Salmonella typhi isolates were multi-drug resistant. The majority of the children were treated with chloramphenicol and ceftriaxone. Typhoid fever appears to be a mild disease in Singapore children. PMID- 9200032 TI - Successful diagnosis and management of cytomegalovirus carditis. AB - Cytomegalovirus (CMV)-associated carditis in the immunosuppressed patient carries a 60% mortality. Underlying pathogenesis is poorly understood but may involve either direct viral invasion or autoimmune cardiac damage triggered in response to the infection. Specific anti-cytomegalovirus therapy and/or anti-inflammatory drugs have been shown to benefit in cases where an early diagnosis was established. We report an unusual case of endo-pericarditis which was temporally related to acute cytomegalovirus infection diagnosed by the immediate early antigen detection in cell culture on whole blood. PMID- 9200033 TI - Usefulness of HCV-RNA assays in efficacy evaluation of interferon treatment for chronic hepatitis C: Amplicor HCV assay and branched DNA probe assay. AB - We investigated the usefulness of Amplicor HCV assay and branched DNA probe assay in efficacy evaluation of interferon therapies for chronic hepatitis C. Subjects were 164 HCV-RNA positive chronic hepatitis C patients who received interferon alpha (IFN-alpha, HLBI) 3-6 MU per day for 14-24 weeks. Their HCV-RNA levels were examined five times by using reverse transcription-polymerase chain reaction (RT PCR) assay, Amplicor HCV assay, and branched DNA probe assay. Complete response rate in the low virus level patients was significantly higher than in the high virus level patients (P < 0.001). In complete responders, the rate of HCV-RNA disappearance 2 weeks after the initiation of IFN therapy was higher than in non responders (P < 0.001). The HCV-RNA positive rates in RT-PCR assay and Amplicor HCV assay agreed by 98% or more. The HCV-RNA negative patients 6 months later were still negative 12 months later by Amplicor HCV assay. Before starting interferon therapy for chronic hepatitis C, it is advisable to make a prediction of treatment efficacy by using branched DNA probe assay. In addition, disappearance of HCV-RNA after 2 weeks of treatment could be a useful predictor of the therapeutic efficacy of IFN. Amplicor HCV assay is useful in detecting HCV RNA and for efficacy evaluation during and after a given interferon therapy. PMID- 9200034 TI - Hepatic pneumocystosis without concomitant PCP in a patient with AIDS. PMID- 9200035 TI - Bilateral facial palsy secondary to herpes zoster meningoencephalitis in a HIV positive woman. AB - Bilateral facial paralysis of diverse infectious aetiologies has been reported in HIV infected patients. We present a patient with bilateral facial palsy most likely due to herpes zoster meningoencephalitis in a patient with neutropenia and who subsequently tested HIV-positive. PMID- 9200036 TI - Pasteurella multocida infection of a total hip arthroplasty following cat scratch. AB - Pasteurella multocida is a well recognized cause of sepsis following animal contact particularly bites and scratches. Spread to prosthetic joints may occur particularly in immunocompromised patients. Immunocompromised patients with prosthetic joints should be warned that animals are potential sources of serious infection and urgent medical advice should be sought if bitten or scratched. PMID- 9200037 TI - The combined value of chemoprophylaxis and pneumococcal vaccine in the prevention of recurrent pneumococcal meningitis. AB - We report the case of an immunocompetent patient who has been the subject of 39 episodes of recurrent pneumococcal meningitis over a 20 year period. The recurrences of bacterial meningitis due to cerebrospinal fluid leakage and the presence of chronic sinusitis were not influenced by the surgical repair of a fistula and the repeated surgical draining interventions on suppurating chronic sinusital foci. Phenoxymethylpenicillin treatment reduced the number of recurrences and the combination of pneumococcal vaccine and penicillin prophylaxis allowed a 5 year period free of any recurrences. PMID- 9200038 TI - Failure of intravenous antibiotic therapy of multiple temporal brain abscesses due to Propionibacterium acnes requiring temporal lobectomy. AB - Propionibacterium acnes is a common skin colonizer. Its involvement in brain abscesses is generally described as a complication of neurosurgical intervention. Propionibacterium acnes is susceptible to antibiotics used as treatment of anaerobic infections, except for the 5-nitroimidazoles. Surgical excision or drainage of a simple abscess combined with a long course of antibiotics is considered the treatment of choice. A case of a patient with multiple brain abscesses located in the right temporal lobe that occurred after the manipulation of an abscess of the right upper maxillary is reported. The patient did not improve despite a prolonged course of high-dose intravenous penicillin plus thiamphenicol and cure was finally obtained after the excision of the right temporal lobe. Culture of the purulent material and the shell of the abscesses yielded P. acnes which was sensitive to all the antibiotics administered to the patient up to the intervention. The temporal lobectomy was followed by a 6-month course of ofloxacin. One year after the intervention, the patient remained apyretic without any other abscess on cranial computed tomography scan. PMID- 9200039 TI - Subclinical rubella reinfection during pregnancy followed by transmission of virus to the fetus. AB - We report a documented case of rubella reinfection during pregnancy in a previously vaccinated woman with residual antibody titre to rubella of 15 IU/ml. The reinfection occurred following an exposure to rubella virus (contact with 6 year-old daughter with clinical rubella) between the 7th and 10th week of pregnancy which resulted in transmission of the virus to the fetus. Umbilical cord blood drawn by cordocentesis was found to be strongly positive for rubella IgM antibody. After termination of the pregnancy rubella virus was isolated in cell culture from fetal tissues. PMID- 9200040 TI - Legionella feeleii pneumonia and pericarditis. AB - Legionella feeleii pneumonia has been described in seven cases, three of them being immunocompromised. We describe a case of L. feeleii pneumonia and pericarditis in a healthy man. Epidemiological survey was not conclusive. To the best of our knowledge, there have been no previous descriptions of pericarditis caused by this organism. PMID- 9200042 TI - Tonsillar actinomycosis presenting as expectorated debris. PMID- 9200041 TI - Staphylococcus aureus bacteraemia, Absidia corymbifera infection and probable pulmonary aspergillosis in a recipient of orthotopic liver transplantation for end stage liver disease secondary to hepatitis C. PMID- 9200043 TI - Cyclospora cayetanensis, potential cause of diarrhoea. PMID- 9200044 TI - Rationalizing back pain: the development of a classification system through cluster analysis. AB - OBJECTIVE: The clinical variations in undifferentiated back pain pose problems for those attempting to develop strategies for care. The objective of this work was to test a methodology for the experimental generation of clinical subgroups of patients with such a complaint, so as to assist more structured study of its natural history and response to treatment. DESIGN: Cluster analysis of dichotomous symptomatic variables from computer-based case histories of three patient cohorts. SETTING: Chiropractic and Orthopedic outpatient clinics. PATIENTS: Three cohorts of new patients with back pain whose symptoms were recorded in a highly standardized way using an interactive computer interview system. CRITERIA ASSESSED: Twenty-four aggravating, relieving and cyclic features of the patients' back complaints assessed for degrees of association and formation of reproducible clusters. RESULTS: Two main patient categories were discerned: one with mechanical features and one that was cyclic. Most patients were assignable to a group. Groupings were largely consistent across all three cohorts and were not related to patient demographics. CONCLUSION: Reproducible and easily-recognized clinical subgroups of back pain patients are possible by cluster analysis using dichotomous case-history variables. More definitive categorization should be obtainable by refining the variable selection and repeating the analysis for additional patient cohorts. These subgroups have the potential to increase the relevance of natural history studies and clinical trials to the day-to-day management of the problem. PMID- 9200045 TI - Chiropractic radiologists: a survey of chiropractors' attitudes and patterns of use. AB - OBJECTIVE: To assess the chiropractic use of radiography, referral patterns to both medical and chiropractic radiologists and attitudes toward radiologists. DESIGN: Random sample mail survey. PARTICIPANTS: Practicing U.S. chiropractors. RESULTS: The response rate was 46% (197 of 425). Seventy-four percent of the respondents have radiographic facilities in their offices. Contraindication screen (71%), pathological diagnosis (63%), biomechanics and posture (51%) and medicolegal protection (27%) were considered important reasons for taking radiographs. When chiropractors refer for radiographic services, 67% refer to medical radiologists and 17% to chiropractic radiologists. Eighty-five percent agreed that chiropractic radiologists are as well qualified as medical radiologists, but 36% thought that medical interpretation carried more legal authority than chiropractic interpretation. Seventy-six percent of respondents thought that the chiropractic radiologist should be consulted only for second opinions. CONCLUSIONS: Most chiropractors obtain radiographs for clinical reasons, such as confirming a diagnosis of pathology, but many continue to use radiography as a screening tool and for medicolegal protection. Prevailing attitudes seem to indicate a need for this specialty in chiropractic, but the chiropractic radiological consultant is not widely used. The disparity between the perceived need for chiropractic radiologists and the current utilization patterns requires further research. PMID- 9200046 TI - Plinth padding and measures of posteroanterior lumbar stiffness. AB - OBJECTIVE: To investigate whether measurement of posteroanterior (PA) lumbar stiffness is affected by the presence of padding on the testing plinth. DESIGN: Within a repeated-measures design, measurements were made of lumbar PA stiffness in subjects without low back pain on both a rigid and a padded plinth surface. SUBJECTS: Nineteen subjects with no history of any low back pain requiring treatment over the preceding 12 months participated in this study. METHODS: PA stiffness was measured at L3 on two occasions under two different conditions. The first condition involved measurement of lumbar stiffness on a rigid plinth surface; the second involved measurement on a padded plinth surface. A reliable mechanical device was used to obtain the PA stiffness measures. RESULTS: Mean lumbar PA stiffness values obtained when testing on a padded plinth were 2.86 N/mm less than those values obtained when testing the same lumbar spines on a rigid plinth. A paired t test showed a significant difference between the PA stiffness measures at L3 obtained on the padded plinth and those obtained on the rigid plinth (t = 6.66, df = 18, p < or = .0001). CONCLUSIONS: These findings suggest that to improve the reliability of lumbar PA stiffness assessment, testing should be performed under the same plinth surface conditions. PMID- 9200047 TI - Supervision of chiropractors: a pilot study. AB - BACKGROUND: Since 1989, Swedish chiropractors with the title "Doctor of Chiropractic" have been authorized as chiropractic physicians by the Swedish health authorities. To obtain authorization after college examination abroad, the new graduate must attend a 1-yr postgraduate course in Sweden. The postgraduate training time in spent half at the hospital (mainly in the orthopaedic department) and half with an authorized chiropractor. OBJECTIVE: To investigate what is important for the chiropractic graduate to learn during his postgraduate training with an authorized chiropractor and which characteristics are important to have to be a good chiropractic supervisor. METHODS: Questionnaires were sent out to 10 authorized chiropractors who have chiropractic graduates at the moment or have had chiropractic graduates in the past. Eleven closed questions and one open question were used. RESULTS: Regarding clinical activities, the most important aspect for the supervisors to learn was to have regular meetings with the graduate. Regarding the graduate's relationship to the patients, the most important aspect was to be professional. In the open question, the most important characteristic to become a good supervisor was to have an interest in teaching. CONCLUSION: Chiropractors feel that one of the most important aspects of supervision is the ongoing dialogue between the chiropractor and the graduate. Among the statements that were considered less important included "awareness of correlation between body and mind" and "to have a holistic approach." It is possible that these two questions received such a low score because chiropractors believe that these activities are not important in the learning process for the graduate. Evaluating what the authorized chiropractor feels is important for the graduate to learn during the postgraduate training will provide some guidelines for those intending to take on graduates in the future. Follow-up studies are needed to compare the opinions of the graduates and the chiropractors. PMID- 9200048 TI - The effect of spinal manipulation in the treatment of cervicogenic headache. AB - PURPOSE: To study whether the isolated intervention of high-speed, low-amplitude spinal manipulation in the cervical spine has any effect on cervicogenic headache. DESIGN: Prospective randomized controlled trial with a blinded observer. SETTING: Ambulatory outpatient facility in an independent research institution. PARTICIPANTS: Fifty-three subjects suffering from frequent headaches who fulfilled the International Headache Society criteria for cervicogenic headache (excluding radiological criteria). These subjects were recruited from 450 headache sufferers who responded to newspaper advertisements. INTERVENTION: After randomization, 28 of the group received high-velocity, low-amplitude cervical manipulation twice a week for 3 wk. The remaining 25 received low-level laser in the upper cervical region and deep friction massage (including trigger points) in the lower cervical/upper thoracic region, also twice a week for 3 wk. MAIN OUTCOME MEASURES: The change from week 1 to week 5 in analgesic use per day, in headache intensity per episode and in number of headache hours per day, as registered in a headache diary. RESULTS: The use of analgesics decreased by 36% in the manipulation group, but was unchanged in the soft-tissue group; this difference was statistically significant (p = .04, chi 2 for trend). The number of headache hours per day decreased by 69% in the manipulation group, compared with 37% in the soft-tissue group; this was significant at p = .03 (Mann Whitney). Finally, headache intensity per episode decreased by 36% in the manipulation group, compared with 17% in the soft-tissue group; this was significant at p = .04 (Mann-Whitney). CONCLUSION: Spinal manipulation has a significant positive effect in cases of cervicogenic headache. PMID- 9200049 TI - George Goodheart, Jr., D.C., and a history of applied kinesiology. AB - Applied Kinesiology (AK), founded by Michigan chiropractor George J. Goodheart, Jr., is a popular diagnostic and therapeutic system used by many health care practitioners. Many of the components in this method were discovered by serendipity and observation. In 1964, Goodheart claimed to have corrected a patient's chronic winged scapula by pressing on nodules found near the origin and insertion of the involved serratus anterior muscle. This finding led to the origin and insertion treatment, the first method developed in AK. Successive diagnostic and therapeutic procedures were developed for neurolymphatic reflexes, neurovascular reflexes and cerebrospinal fluid flow from ideas originally described by Frank Chapman, D.O., Terrence J. Bennett, D.C., and William G. Sutherland, D.O., respectively. Later, influenced by the writings of Felix Mann, M.D., Goodheart incorporated acupuncture meridian therapy into the AK system. Additionally, the vertebral challenge method and therapy localization technique, both based on phenomena proposed by L. L. Truscott, D.C., were added to the AK system. Scholarship has also evolved regarding AK and research on the topic is in its infancy. This paper documents some of the history of AK. PMID- 9200050 TI - Manipulative reduction and management of anterior sternoclavicular joint dislocation. AB - OBJECTIVE: To discuss the management of a patient suffering from an anterior sternoclavicular joint dislocation secondary to blunt trauma from a motor vehicle accident. CLINICAL FEATURES: A 75-yr-old woman suffered from difficulty in swallowing and chest pain after release from a foreign medical facility. Anteroposterior chest X-rays demonstrated an anterior and superior displacement of the right sternoclavicular joint. INTERVENTION AND OUTCOME: Specific joint manipulation for reduction of the dislocation was performed. Immobilization of the joint after reduction was accomplished by a reverse figure-8 bandage. Follow up radiographic evaluation demonstrated reduction of the dislocation. Resolution of difficulty in swallowing and pain was dramatic and instantaneous after reduction. CONCLUSION: Appropriate intervention of chiropractic examination procedures and imaging techniques culminated in successful resolution of this case. When such cases are recognized, appropriate management may occur conservatively with judicious application of joint manipulation and adjunctive procedures. PMID- 9200051 TI - The association between visual incompetence and spinal derangement: an instructive case history. AB - OBJECTIVE: To discuss the difficulties encountered in the diagnosis of concentric narrowing of the visual fields. CLINICAL FEATURES: A 13-yr-old child was referred to a chiropractic clinic after an ocular examination concerning a 6-month period of minor headaches, which culminated in a more severe attack, during which she had to lie down. Her examination was essentially normal, with the exception of the presence of constricted visual fields when measured to a small stimulus. INTERVENTION AND OUTCOME: Standard outpatient spinal adjustments were followed by recovery of vision. DISCUSSION: The recovery of constricted fields of vision with spinal manipulation has now been discussed with greater frequency in the chiropractic literature. The diagnosis of constriction of the visual fields is often the factor that may decide the further management of the patient. In this instance, constriction of the visual fields could easily have been missed, even though clinical examination of the visual fields had been done. CONCLUSION: Elements in the history and physical examination will suggest when a sensitive assessment of the visual fields may be of benefit to the patient. PMID- 9200052 TI - Why the term "carrier screening" should be abandoned. PMID- 9200053 TI - Epidemiological clues from screening. PMID- 9200054 TI - Excess mortality from breast cancer in relation to mammography screening in northern Sweden. AB - OBJECTIVES: Previous randomised studies of mammography screening have shown a significant effect on breast cancer mortality, particularly in women aged 50-69 at randomisation. Breast cancer mortality has traditionally been studied by judgments on causes of death, either from cause of death registers or from medical records. In this study an alternative method was used, estimating the excess mortality associated with breast cancer. SETTING: In 1990 two counties of northern Sweden started population based mammography screening of women aged 40 74. The unscreened population in the two other counties of the same region were selected as controls. RESULTS: Excess mortality associated with breast cancer was lower in the screened population, and was discernible three to four years after the start of screening. The relative risk estimate, based on the cumulative excess number of deaths from breast cancer during 1990-95 in the screened versus the control population aged 40-74 (at diagnosis of breast cancer), was 0.72 (95% confidence interval (CI) 0.53 to 0.99). For women aged 50-69 it was 0.67 (95% CI 0.46 to 0.99). In the 50-69 age group the estimated excess number of deaths from breast cancer during 1995 was 17.0 per 100,000 women (95% CI 5.0 to 29.0) in the screened counties and 51.1 per 100,000 (95% CI 30.2 to 71.9) in the unscreened counties. CONCLUSIONS: Population based routine screening has substantial effects on breast cancer mortality in women aged 50-69. Estimation of excess mortality can be used in future studies to evaluate the effects of mammography screening on breast cancer mortality. PMID- 9200055 TI - UK breast screening programme: how does it reflect the Forrest recommendations? AB - OBJECTIVE: To compare the UK breast screening programme with the Forrest Report recommendations of 1986. SETTING: The UK breast screening programme. METHODS: A postal survey of 97 local breast screening programmes in the United Kingdom. The main outcome measures were the frequency of screening, the use of two view screening on incident screens, reading of screening mammograms, assessment procedures and visits, staffing levels, and the use of building and equipment. RESULTS: Eighty two (85%) of the questionnaires were completed and returned. All programmes screen every three years, as Forrest intended, with the exception of one health region which screens more often. The national policy is to use two views on incident screens where there is a clinical indication. None the less, 14% of programmes are using, or intending to use, two views on all women. Double reading of mammograms is not recommended in the United Kingdom outside Scotland, but is used by 88% of programmes. All programmes have access to the equipment required for the assessment techniques recommended by Forrest. Variation exists between programmes in the procedures women can expect to receive at their initial assessment visit and in the total number of assessment visits. Sixty eight per cent of programmes' breast screening budgets cover the staff required for a multidisciplinary team as defined by the Forrest Report. Ninety three per cent of screening programmes are organised around static sites, with 86% of these also using mobile vans. CONCLUSIONS: The national programme is following recommendations about the frequency of screening, but there seems to be some divergence from policy as regards the use of double reading, two views at incident screening, and the multidisciplinary team covered by the programmes' breast screening budget. Further research is needed on the effectiveness and cost effectiveness of two view incidence screening, double reading, and non radiologists as readers. Investigation is also needed of the costs and effects of the variation between programmes in the number of assessment visits a woman may have. PMID- 9200056 TI - Breast cancer screening: the effect of self selection for screening on comparisons of randomised controlled trials. AB - In randomised controlled trials of breast cancer screening women are randomly allocated to an intervention group that is offered screening, or a control group that receives the usual medical care. Not all women in the intervention group accept screening; the women who do so may differ from other women in their underlying risk of breast cancer. This self selection for screening can result in either women at high risk or at low risk being overrepresented in the screened group. When comparisons between trials are made it is important to take self selection for screening into account, as a trial with self selection of women at high risk has the potential to have greater efficacy than a trial with self selection of women at low risk. A method of adjusting for self selection when comparing randomised controlled trials of breast cancer screening is described. PMID- 9200057 TI - Impact of follow up letters on non-attenders for breast screening: a general practice based study. AB - OBJECTIVE: To determine the effectiveness of follow up letters to non-attenders for screening on the breast screening uptake in practices with a low preliminary uptake of screening. DESIGN: Observational study of two groups of general practices. In 40 of these practices, the preliminary uptake of screening was less than 60%. These 40 practices were offered help from a clerical officer to check names and addresses of non-attenders, and to send non-attenders a reminder letter. SETTING: 93 general practices in South West London in 1995-96. MAIN OUTCOME MEASURES: Preliminary and final uptake of breast screening. RESULTS: Breast screening uptake increased by an average of 4.6% in the 40 intervention practices compared with 1.6% in the 53 control practices (difference 3.0%, P < 0.0001). However, the absolute increase in the uptake of screening in the intervention group was small (from 53.8% to 58.5%). The marginal cost for each additional women screened was Pounds 7 (compared with an average cost for each women screened of Pounds 27). CONCLUSIONS: Reminder letters can help increase the uptake of screening in practices with a low preliminary uptake of breast screening. However, they have a limited role in improving the uptake of breast screening in inner city areas, and other methods of increasing uptake therefore need to be developed and evaluated. PMID- 9200058 TI - Presentation of screen negative results on serum screening for Down's syndrome: variations across Britain. AB - OBJECTIVES: To document current practice of communicating screen negative results to pregnant women undergoing a test for Down's syndrome. SETTING: 169 British NHS hospital antenatal clinics currently offering multiple marker serum screening for Down's syndrome and giving results directly to women. METHODS: All 169 clinics were sent a letter asking about the method and form of communicating screen negative results. RESULTS: In only 29% of programmes were specific arrangements made to inform women of screen negative results, and in 5% these results were not given at all. Screen negative results were given as a verbal phrase in 44% of programmes, as a risk figure in 16% of programmes and as both in 40% of programmes. CONCLUSIONS: These results highlight a gap between screening policy guidelines and practice in the case of Down's syndrome serum screening. PMID- 9200059 TI - Neonatal screening for cystic fibrosis in the Trent region (UK): two-stage immunoreactive trypsin screening compared with a three-stage protocol with DNA analysis as an intermediate step. AB - OBJECTIVES: To assess neonatal screening for cystic fibrosis using immunoreactive trypsin, either alone or in conjunction with DNA analysis for the delta F508 mutation. A novel three-stage screening protocol was compared with the previously introduced two-stage immunoreactive trypsin-DNA protocol. DESIGN: (a) Collection of data from a 4 1/2 year period (phase 1) of two-stage immunoreactive trypsin screening. The initial dried blood samples were obtained at 6 days of age and repeat samples at 27 days of age from babies with results above the 99.5th centile. Babies with persistent hypertrypsinaemia were referred for a diagnostic sweat test. (b) Retrospective DNA analysis: patients with cystic fibrosis diagnosed in phase 1 were genotyped and most samples from babies with increased initial immunoreactive trypsin but normal results in the second sample were analysed for the delta F508 mutation. (c) Phase 2, a prospective study of a three stage neonatal screening protocol, in which only babies heterozygous for the delta F508 cystic fibrosis mutation progressed to the second immunoreactive trypsin test. SETTING: The Trent neonatal screening programme. SUBJECTS: 437 859 babies born between August 1989 and March 1996. MAIN OUTCOME MEASURES: Proportions of unaffected babies requiring a second blood sample or a sweat test. Overall sensitivity for the detection of cystic fibrosis. RESULTS: The two-stage screen failed to identify six out of 94 cases of cystic fibrosis (without meconium ileus). The introduction of the DNA analysis step would have resulted in one additional case being missed. With the three-stage screen there was a 92% reduction in babies requiring a second blood sample and an 80% reduction in negative sweat tests, results close to the predictions of the retrospective study. CONCLUSIONS: The three-stage screening protocol is a marked improvement on the two-stage immunoreactive trypsin strategy and on the two-stage immunoreactive trypsin-DNA strategy recently introduced in some other screening programmes. PMID- 9200060 TI - A survey of screening compliance among first degree relatives of people with colon cancer in New South Wales. AB - OBJECTIVE: To survey screening practices, knowledge, and attitudes towards screening among first degree relatives of people with colon cancer. SETTING: A random sample of people with colon cancer listed on the New South Wales (NSW) Cancer Registry were mailed a questionnaire to be passed on to an appropriate first degree relative. METHODS: Two hundred and twenty five first degree relatives completed a self administered questionnaire. RESULTS: Although there were high levels of awareness about colorectal cancer, and attitudes towards colorectal cancer were generally positive, screening rates were low, and only three relatives had been screened in accordance with current Australian recommendations. Factors associated with previous participation in any type of screening test (usually once) included receiving a medical recommendation to screen, having more than one relative with colorectal cancer, being a sibling of the relative with colon cancer, the relative with cancer being female, and perceiving screening as messy, but not painful. CONCLUSIONS: Strategies to enhance screening awareness and participation among relatives need to be considered. This study provides some insight into factors to be considered in developing awareness programmes. Further research is required to explore these factors, and to identify ways to overcome barriers. PMID- 9200061 TI - General practitioner based screening for cervical cancer: higher participation of women with a higher risk? AB - OBJECTIVE: To test the hypothesis that a personal invitation for cervical screening by a woman's own general practitioner (GP) achieves a higher attendance of women with an increased risk for cervical cancer. SETTING: Two general practices and the local health authority screening programme for cervical cancer, Nijmegen, The Netherlands. METHODS: Attendance rates of women with an increased risk of cervical cancer were compared for two invitation strategies: (a) invitation by the woman's own GP, and (b) invitation by a national call system through the local health authority. Data on risk profiles were gathered by questionnaire. Two hundred and thirty eight women eligible for screening were invited by their GPs (GP group), and 235 women by the local health authority (control group) in 1992. RESULTS: The personal invitation by the GP resulted in an 18% higher overall attendance, and a 28% higher attendance of women with greater risk because of sexual behaviour and smoking. CONCLUSION: Greater involvement of the GP in inviting women for cervical cancer screening results in a higher attendance, particularly among women with increased risk, than a less personal health authority call system. PMID- 9200062 TI - Changing incidence of invasive adenocarcinoma of the uterine cervix in East Anglia. AB - OBJECTIVE: To determine trends in incidence of invasive adenocarcinoma of the uterine cervix in East Anglia. METHODS: Cervical cancer incidence data for both squamous cell carcinomas and adenocarcinomas were obtained from the East Anglian Cancer Registry for the period 1971-94. Similar data were obtained for England and Wales. European age standardised rates (ASRs) were used for comparisons. RESULTS: The mean incidence (ASR) of cervical adenocarcinoma was 0.85 per 10(5) in 1971-76, rising to 2.54 per 10(5) in 1989-94. There has been a marked age shift, with the main increase in incidence occurring in younger women aged 30-39. The mean incidence (ASR) of squamous cell carcinoma of the cervix has decreased from 9.78 to 8.74 per 10(5) over the periods 1971-76 and 1989-94. Again there has been an age shift, moving from a single incidence peak in the 45-59 age band in earlier years to incidence peaks in both the 30-39 and 55-69 age bands in more recent years. Similar trends were noted when data for England and Wales were analysed. Birth cohort analyses show that both tumours are occurring progressively earlier (about five years earlier in each five year birth cohort). CONCLUSION: Although the overall incidence of cervical carcinoma is declining, this study has shown an increased incidence of cervical adenocarcinoma, particularly in the younger age groups. In future it would seem advisable to publish separate incidence and mortality data for squamous cell carcinoma and adenocarcinoma of the uterine cervix. All practitioners involved in the cervical cancer screening programme would then be aware of the very real significance of this tumour. PMID- 9200063 TI - Trends in invasive cervical cancer incidence in East Anglia from 1971 to 1993. AB - OBJECTIVE: To study the trends in the incidence of invasive cervical cancer in East Anglia. DESIGN: Statistical analysis of age specific incidence rate for the period 1971-93 using East Anglian Cancer Registry data. SUBJECTS: All cases of invasive cervical cancer registered with the East Anglian Cancer registry, diagnosed in the period 1971-93. MAIN OUTCOME MEASURES: Changing incidence of cervical cancer. RESULTS: For the 20 years 1971-90, trends varied widely by district and by age group, with little discernible overall effect of the increasing screening activity. Since 1990, rates have fallen sharply in the age groups targeted for screening, with a reduction of 34% (95% confidence interval 26% to 42%) from that expected based on 1971-90 trends. This fall was preceded by a rapid rise in the national uptake of screening. A shift to more favourable stage at diagnosis has also occurred. CONCLUSION: Changes in the organisation and management of the national screening programme introduced in 1988 and 1989 seem to have led to substantial improvements in effectiveness. PMID- 9200064 TI - Quality of the measurement of the infrarenal aortic diameter by ultrasound. AB - OBJECTIVES: To assess quality and variability in measurements of the infrarenal aortic diameter by ultrasound, and to recommend quality control measures to improve consistency in measurements of the infrarenal aortic diameter (IAD) in a long running screening programme. SETTING: An aneurysm screening programme in Huntingdon. METHODS: Quality of the ultrasound image was subjectively assessed by each ultrasonographer. Quality of the measurements was assessed by analysing the frequency of measurements that were outside the normal variability of the estimated true diameter. The interobserver variability was measured by analysing repeated measurements of the IAD in the same patient by two ultrasonographers, using the same scanner. The variability between different scanners was measured by analysing repeat measurements of the IAD in the same patient by the same ultrasonographer, using two scanners. The intraobserver variability was estimated by analysing all patients with three consecutive measurements of the IAD, carried out by the same ultrasonographer. RESULTS: Although the subjective assessment of the quality of the ultrasound image of the aorta varied, there were no statistically significant differences in the likelihood of obtaining measurements outside the limits of agreement between the ultrasonographers. The interobserver, intraobserver, and between scanner variability of ultrasound measurements of the IAD were all around 6 mm. CONCLUSION: The variability in ultrasound measurements of aortic diameters is acceptable for clinical decision making, and the interobserver variability is very similar to the long term intraobserver variability. Quality control measures are suggested to maintain long term consistency of ultrasound measurements of the IAD. PMID- 9200065 TI - An oligopeptide of the feline leukemia virus envelope glycoprotein is associated with morphological changes and calcium dysregulation in neuronal growth cones. AB - Neuropathogenic processes that affect the pathfinding properties of neuronal growth cones could account for many of the dysfunctions unique to retroviral infection of developing nervous systems. Pediatric HIV-1 infection, for example, is associated with a distinctive neuropathogenesis that includes marked cortical atrophy, cognitive disorders, and pyramidal dysfunction. The ability of HIV's envelope glycoprotein, gp120, to produce increased intracellular free calcium ([Ca2+]i) leading to neuronal death has been documented. We hypothesize that gp120 and the envelope glycoproteins of other retroviruses may have similar calcium-increasing effects in advancing growth cones, a property which could disrupt the orderly development of the nervous system. To explore this possibility, we exposed chick ciliary ganglion neurons in culture to a known cytopathic region (CVR5) of the feline leukemia virus' envelope glycoprotein. CVR5 produced [Ca2+]i increases and dose-dependent morphological changes in growth cones isolated from their cell bodies by axotomy. These responses of growth cones to CVR5 suggest that the neurotoxic effects of retroviruses could be mediated at the level of the individual growth cone through exposure to envelope glycoproteins and could constitute one mechanism by which these viruses perturb the normal development of the nervous system. PMID- 9200066 TI - Search for herpesvirus DNA in cerebrospinal fluid of HIV patients with brain disorders: prevalence of cytomegalovirus DNA findings. AB - A study was carried out to search for the presence of the seven human herpesvirus DNAs in cerebrospinal fluid from 52 human immunodeficiency virus-infected patients with brain disorders. Cytomegalovirus DNA was the most prevalent with 12 positive samples; Epstein-Barr virus and varicellazoster DNAs were detected in three and two samples, respectively, while no sample was positive for the DNA of the other herpesviruses. PMID- 9200067 TI - Differences in resistance to herpes simplex virus type 1 (HSV-1) among oligodendroglia derived from different strains of mice are determined after viral adsorption but prior to the expression of immediate early (IE) genes. AB - The nature of an innate cellular resistance to HSV-1 of cultured murine oligodendrocytes (OLs) in three strains of mice (C57BL/6J, Balb/cByJ and A/J) was investigated. The expression of immediate early (ICP4), early (ICP8) and late (gC) antigens in primary OL cultures was studied using an indirect immunofluorescence (IF) technique. HSV-1 infected OLs from C57BL/6J mice showed no viral antigens at 24 h post infection (p.i.) but rather a marked delay in antigen expression beginning at 60 h p.i. In contrast all three proteins were expressed in A/J OLs at 24 h p.i. while Balb/cByJ OLs showed an intermediate protein expression pattern. These results suggest that the innate cellular resistance to HSV-1 is determined prior to the expression of immediate early viral antigens. To further study these differences, the adsorption capacity between the three mouse strains was compared using dextran purified, [3H]thymidine labelled virus. No differences in HSV-1 adsorption were identified. Results from viral penetration studies approached statistical significance suggesting that penetration may be impaired in C57BL/6J and Balb/cByJ OLs when compared to A/J OLs and is likely fusion independent. The selective differences in HSV-1 resistance mediated by OLs, reflect differences in virus host cell interactions, that likely contribute to differences in mortality, viral spread, and the ability of virus to induce central nervous system (CNS) demyelination. PMID- 9200068 TI - Nerve growth factor antibody stimulates reactivation of ocular herpes simplex virus type 1 in latently infected rabbits. AB - Anti-nerve growth factor (anti-NGF) antibody has been shown to induce reactivation of latent herpes simplex virus type 1 (HSV-1) in vitro. We found that systemically administered anti-NGF induces ocular shedding of HSV-1 in vivo in rabbits harboring latent virus. Rabbits in which HSV-1 latency had been established were given intravenous injections of goat anti-NGF serum daily for 10 days beginning 42 days after primary viral infection. Tears were assayed for virus for 12 days beginning on the day of the first injection. All eight rabbits given high titer anti-NGF had infectious virus in their tears at least once during the 12-day period. Fifteen of 16 eyes were positive and the average duration of viral shedding for these eyes was 4.0 days. Latently infected rabbits receiving daily injections of nonimmune goat serum or saline for 10 consecutive days were controls. Only six of the 16 (38%) eyes from rabbits receiving nonimmune goat serum shed virus. Only one of 12 eyes from untreated rabbits shed virus. Sera from control rabbits had no detectable anti-NGF activity; titers in anti-NGF-treated rabbits ranged between 1:1000 and 1:10,000. NGF deprivation may act as a neuronal stressor and may share a common second messenger pathway with heat- or cold-stress induced reactivation of latent HSV-1. PMID- 9200069 TI - Repression of the HSV-1 latency-associated transcript (LAT) promoter by the early growth response (EGR) proteins: involvement of a binding site immediately downstream of the TATA box. AB - During herpes simplex virus (HSV) latency, in neurons of the nervous system, a single family of viral transcripts (the Latency-Associated Transcripts or LATs) are synthesized. Within the LAT promoter region, we have identified a consensus sequence for the EGR proteins in an unusual position immediately downstream of the TATA box. The early growth response (EGR) proteins are rapidly induced in cells by stimuli which also induce HSV to reactivate from latency. In order to determine if EGR proteins play any role in control of LAT transcription, we have analyzed the interactions between EGR proteins and the LAT promoter. Gel retardation and DNase I protection assays demonstrated that EGR1 zinc finger protein bound specifically to the LAT promoter region EGR consensus sequence. To determine if EGR proteins could modulate transcription through the LAT promoter, cotransfection assays were performed using chloramphenicol acetyltransferase (CAT) reporter constructs driven by either the wild-type LAT promoter or a LAT promoter with a mutated EGR binding site. Contransfection of the wild-type LAT promoter construct with EGR expression plasmids resulted in inhibition of the basal level of CAT activity with EGR-2 but not EGR-1 or 3. However, normal levels of CAT activity were observed in cotransfections using the mutant LAT promoter CAT construct suggesting that repression was mediated by the binding of EGR-2 proteins to the LAT promoter. Furthermore, data from combination binding assays using EGR1 and TATA binding protein (TBP) in vitro support the hypothesis that binding of EGR proteins to the LAT promoter prevents binding of TBP and thus suppresses transcription. These results may provide a link between stress responses in neurons of the CNS which activate the EGR family of proteins and HSV reactivation from latency due to the same stress response. PMID- 9200070 TI - CD4+ and CD8+ T cells are not major effectors of mouse hepatitis virus A59 induced demyelinating disease. AB - We examined murine hepatitis virus strain A59 (MHV-A59)-induced demyelinating disease in C57BL/6 mice which had previously been thymectomized at 25 days of age. Demyelination was observed in 51-96% of spinal cord quadrants examined 30 or 60 days post infection (dpi), indicating that neither an intact thymus nor thymic infection is a prerequisite to demyelination. Depletion of CD4+ or CD8+ T cells at 5, 7 or 10 dpi did not influence the extent of demyelination indicating that neither T cell subset is a major effector of demyelination. However, these findings do not exclude the possibility that T cells are involved in initiating demyelinating disease very early in infection. PMID- 9200071 TI - Primary cerebellar T-cell lymphoma with acquired immunodeficiency syndrome. AB - We report a 33-year-old man with the acquired immunodeficiency syndrome (AIDS) and a solitary T-cell lymphoma. Systemic sites of lymphomatous involvement could not be identified. Subtotal resection of the lesion with cranial irradiation resulted in a marked neurologic improvement. Our case suggests that T-cell lymphomas should be considered in the differential diagnosis of a solitary mass of the cerebellum in patients with AIDS and that aggressive therapy may be warranted. PMID- 9200072 TI - Stable neurological function in subjects treated with 2'3'-dideoxyinosine. AB - AIDS Dementia Complex (ADC) is a frequent and devastating complication of HIV infection. There is evidence that zidovudine (ZDV) has an effect in alleviating the symptoms of ADC, and may have a role in its prevention. It is therefore important that new antiretroviral therapies be evaluated not only for the risk of neurologic side effects, but also for their relative efficacy to ZDV in the prevention of ADC. The present study reports the effects of 2'3'-dideoxyinosine (DDI, didanosine, Videx) therapy on neuropsychological performance in the context of several large clinical trials targeting advanced systemic HIV-1 infection. Subjects treated with DDI had stable neurologic performance in quantitative tests over a 1 year period and were similar to zidovudine treated subjects. PMID- 9200073 TI - Pediatric surgeons' activities and future plans. AB - OBJECTIVE: The authors aim was to survey members of the Canadian Association of Paediatric Surgeons (CAPS) on their demographics, practice, and future plans. MATERIALS AND METHODS: A questionnaire was mailed to 86 members. RESULTS: We received 60 questionnaires (70%), with a return rate including 85% men and 15% women. Seventeen percent of the respondents do not have children, 8% did not answer the question pertaining to children, and 8% expect to have more children. Among the 75% of surgeons with children, 16% have reduced the number of hours worked, from 2 hours to more than 1 day per week, for a number of years. Younger surgeons and women are more likely to reduce their work load for their family life. Most surgeons practice in an academic (64%) or a mixed setting (25%), with only 12% involved in private practice. On average, 69% of their time is devoted to patient care; teaching and research each take an average of 10% of the surgeon's time, while 9% of their time is spent on administrative duties. Study respondents work an average of 57 hours per week, and 45 weeks per year. Age significantly influenced the number of hours worked per week, and the number of weeks worked on a yearly basis. Gender and type of practices did not significantly influence the number of hours or weeks worked, whereas location of practice did. Spouse activity also had an impact on the number of hours and weeks worked. When asked about their preference for the next 5 years, 30% of surgeons would opt for a decrease in their level of activity, and 15% wish to retire. Irrespective of the age group or the type of practice, surgeons would prefer to decrease their level of activity. A recurring theme submitted by respondents is the need for increased time for teaching and research and less administrative work. Finally, 60% of surgeons were very satisfied with their work, 27% were satisfied, and 12% were unsatisfied. CONCLUSION: Lifestyle and family commitment have an impact on pediatric surgeons' activity and should be considered when analyzing work force requirements. PMID- 9200074 TI - Histopathologic analysis of interval appendectomy specimens: support for the role of interval appendectomy. AB - The treatment of appendiceal abscess is controversial. For patients initially treated "conservatively" with antibiotics with or without drainage, the role of interval appendectomy is an area of considerable debate. Without interval appendectomy, the true risks of recurrent disease and missed pathological findings are uncertain, and large, long-term, prospective studies are unavailable. To evaluate the role of interval appendectomy, the authors reviewed the histopathologic specimens from patients with presumed appendiceal abscess treated by interval appendectomy. Over a 7-year period, 162 children presented with a clinical diagnosis of perforated appendicitis. Eighteen patients had localized abscesses treated conservatively, followed by interval appendectomy. Standard histopathologic sections of 17 of the 18 appendices were examined by one pathologist who was blinded to the clinical data and to the interpretation of the original pathologist. Of the 11 boys and seven girls (mean age, 7.4 +/- 3.4 years), eight underwent percutaneous drainage and one underwent operative drainage. All received intravenous antibiotics for a mean of 8.6 +/- 3.2 days with a hospital stay of 10.4 +/- 8.3 days. Interval appendectomy was performed at a mean of 92.7 +/- 20.7 days after initial admission, with discharge at a mean of 2 +/- 1.3 days after surgery. There were no complications or deaths. Histopathologic review showed normal appendix (n = 4), normal appendix with mild serositis (n = 6), normal appendix with unsuspected resolved Meckel's diverticulitis (n = 1), appendiceal duplication (n = 1), granulomatous appendicitis (n = 3), and acute appendicitis (n = 2). All appendices had patent lumens, and 15 were documented to be present to the tip. There was no correlation between the histopathologic findings and the interval between abscess and interval appendectomy. Interval appendectomy was performed with no morbidity and a short hospital stay. Two patients had histopathologic recurrent acute appendicitis, five had unsuspected pathological findings (appendiceal duplication, Meckel's diverticulitis, granulomatous inflammation), and none of the appendices had an obliterated lumen, suggesting that all patients were at long-term risk for recurrent disease. These data support the role of interval appendectomy in cases of perforated appendicitis treated conservatively. PMID- 9200075 TI - Hemangioma of the umbilical cord mimicking an omphalocele. AB - A hemangioma of the umbilical cord was misdiagnosed as an omphalocele both antenatally and immediately after birth. The girl was full term and had no other anomalies. Complete resection, including an intraabdominal component, was performed on the first day of life. This rare anomaly and its possible pitfalls are discussed, and the literature is reviewed. PMID- 9200076 TI - Effect of major surgery on neutrophil chemotaxis and actin polymerization in neonates and children. AB - The authors have examined the effect of major surgery in neonates and older children on neutrophil (PMN) chemotaxis and on actin polymerization, an essential early step in PMN movement. Isolated PMNs from the following subjects were studied: healthy adult volunteers (n = 28), healthy newborns (n = 21), newborns undergoing major surgery (n = 7), and older infants and children undergoing major surgery (n = 14). Chemotaxis was measured by a micropore filter assay, and actin polymerization was measured by flow cytometry. Blood samples from surgical patients were obtained preoperatively, hourly during the procedure, immediately postoperatively, and 48 hours after surgery. Mean preoperative newborn PMN chemotaxis was similar to that of healthy newborn PMN, and mean preoperative PMN chemotaxis in children was similar to that of healthy adults. There were no significant alterations in PMN chemotaxis during or after major surgery in neonates or children. Peak PMN actin polymerization, after stimulation with formyl methionyl leucyl phenylalanine (FMLP) (10 nm), was significantly diminished in healthy neonates compared with adults (P < .005). Preoperative surgical neonates had similar peak PMN actin polymerization levels to those of healthy newborns, and older preoperative children had similar levels to adults. PMN actin polymerization did not significantly change during or after major surgery. Despite reductions in PMN chemotaxis and actin polymerization in healthy neonates, there is no further impairment of these PMN functions during or after major surgery. Our data suggest that PMN chemotactic function is resistant to the stress of uncomplicated major surgery in neonates and children. PMID- 9200077 TI - Does extracorporeal membrane oxygenation benefit neonates with congenital diaphragmatic hernia? Application of a predictive equation. AB - The overall survival of neonates with congenital diaphragmatic hernia (CDH) remains poor despite the advent of extracorporeal membrane oxygenation (ECMO). Attempts at accurately predicting survival have been largely unsuccessful. The purpose of this study was twofold: (1) to identify independent predictors of survival from a cohort of CDH neonates treated at the authors' institution when ECMO was not available and combine them to form a predictive equation, and (2) to apply the equation prospectively in a cohort of CDH neonates, treated at the same institution when ECMO was available, to determine whether ECMO improves outcome. From the clinical data of 62 CDH neonates treated at the authors' center by the same team of university neonatologists and pediatric surgeons between 1983 and 1993 (before ECMO availability), 15 preoperative and seven operative variables were selected as potential independent predictors. When subjected to multivariate, stepwise logistic regression analysis, four variables were identified as statistically significant (P < .05), independent predictors of survival: (1) ventilatory index (VI), (2) best preoperative PaCO2, (3) birth weight (BW), and (4) Apgar score at 5 minutes. When combined via logistic regression analysis, the following predictive equation was formulated: P (probability of survival to discharge) = [1 + e(x)]-1 where x = 4.9 - 0.68 (Apgar) - 0.0032 (BW) + 0.0063 (VI) + 0.063 (PaCO2). Applying a standard cut-off rate of survival at less than 20%, the equation yielded a sensitivity of 94% and a specificity of 82% in identifying the correct outcome of patients treated with conventional ventilatory management. The overall survival rate was 66%. Since the availability of ECMO at the center, 32 CDH neonates were treated using the same conventional ventilatory treatment and surgical repair by the same university staff. The overall survival rate was 69%. The predictive equation was applied prospectively to all neonates to determine predicted outcome, but was not used to decide the treatment method. Eighteen neonates received conventional therapy alone; 16 of 18 survived (89%). Fifteen of the 16 patients who survived had their outcomes predicted correctly (94%). Fourteen neonates did not respond to conventional therapy and required ECMO; 6 of 14 survived (43%). Six of the eight patients predicted to survive, lived (75%). All six patients predicted to die, died despite the addition of ECMO therapy (100%). The mean hospital cost, per ECMO patient who died, was $277,264.75 +/- $59,500.71 (SE). An odds ratio analysis, using the four independent predictors to standardize for degree of illness, was performed to assess the risk associated with adding ECMO therapy. The result was 1.25 (P = 0.75). Although the cohort was not large enough to eliminate significant beta error, the data strongly suggested no advantage of ECMO. At this center, absolute survival rates for neonates with CDH have not been significantly altered since ECMO has become available (66% v 69%). The authors conclude that the predictive equation remains an accurate measurement of survival at their center even when ECMO is used as a salvage therapy. The method of creating a predictive equation may be applied at any institution to determine the potential outcome of CDH neonates and assess the effect of ECMO, or other salvage therapies, on survival rates. PMID- 9200078 TI - Do children with repaired low anorectal malformations have normal bowel function? AB - PURPOSE: The authors' aim was to compare bowel function in patients who had undergone repair of a low anorectal malformation with that of normal healthy children. METHODS: The bowel function of 40 patients (29 boys, 11 girls; median age, 7; range, 3 to 13) with low anorectal malformations was evaluated by a multivariate scoring method based on a questionnaire. All patients were toilet trained for defecation and micturition. They were also evaluated clinically, and the outcome was graded as excellent (normal bowel function), good (no or minor social limitations), fair (marked social limitations), or poor (total incontinence). Fifty-four healthy children with a similar age and gender distribution were used as controls. RESULTS: Twenty-one patients (52%) with normal bowel function had continence scores within the range of the scores of healthy children (patients, 19.3 +/- 0.7 v controls, 19.1 +/- 1.3). Fifteen patients had a good clinical outcome. The mean score in this group was 16.3 +/- 2.4. Four patients with a fair outcome had a mean score of 10.5 +/- 2.9. Constipation requiring dietary or medical treatment was reported by 17 patients (42%) and four of the controls (7%). Daily soiling caused by fecal overflow was reported by four patients (10%) and none of the controls. None of the patients had urinary incontinence; occasional wetting was found in 27% of the patients and 22% of the controls (P = .56). CONCLUSION: Only half of the children with a low anorectal malformation have age-appropriate normal bowel function. Long-term follow-up of these patients to manage the main functional problem, constipation, is warranted. PMID- 9200080 TI - Capsulectomy: a cure for the page kidney. AB - Hypertension is a known complication after renal trauma. The cause of posttraumatic hypertension can be renal scarring, infarction, hydonephrosis, infection, vascular injury, and parenchymal compression. The authors report on the case of a 16-year-old boy who experienced hypertension after blunt renal trauma. He had a dense fibrous pseudocapsule causing renal parenchymal compression, which lead to hypertension, a Page kidney. Evaluation with computed tomographic (CT) scan, radioisotope renal scan, renal Doppler, and angiogram confirmed the diagnosis. Removal of the renal capsule and the constricting fibrous pseudocapsule was curative. PMID- 9200079 TI - Collagen induces cytokine release by fetal platelets: implications in scarless healing. AB - In previous studies the authors demonstrated that unlike adult platelets, fetal platelets respond poorly to collagen. When platelets make contact with the exposed collagen at the site of injury, the result is activation, aggregation, and degranulation with the release of cytokines and growth factors. This sequence of events is well characterized in adult wounds, which heal with an acute inflammatory response and dense scar formation. In sharp contrast, fetal dermal wounds heal without an acute inflammatory response and minimal scar formation. Therefore, the aim of this study was to test the hypothesis that collagen, abundant at the site of dermal injury, is a poor inducer of cytokine release by fetal platelets. This could explain, in part, the minimal inflammation accompanying fetal dermal wound healing. Platelet suspensions from six fetal Yorkshire swine at day 80 of gestation (term, 114 days) were exposed to either arachidonic acid, 0.5 mg/mL, collagen, 0.19 mg/mL, or saline. The release into plasma of transforming growth factor-beta (TGF-beta 1), and platelet-derived growth factor (PDGF)-AB effected by these agents was determined by enzyme-linked immunosorbent assays. Transmission electron microscopy (TEM) was used to correlate the biochemical findings with ultrastructural changes and showed that arachidonate-treated platelets were aggregated and devoid of granules. In contrast, collagen-treated platelets had undergone conformational changes but showed only a moderate change in the quantity and homogeneity of their secretory granules compared with saline-treated controls. There was a significant increase in TGF-beta 1 release into plasma after treatment with collagen (6.64 +/- 0.36 ng/mL) and arachidonate (7.64 +/- 0.77 ng/mL) compared with saline (4.74 +/- 0.36 ng/mL), P < .05. Likewise, PDGF-AB release was significantly higher after collagen (0.22 +/- 0.02 ng/mL) and arachidonate treatment (0.44 +/- 0.04 ng/mL) compared with saline (0.09 +/- 0.02 ng/ mL), P < .05. The authors conclude that fetal platelets actually do release cytokines in response to contact with collagen despite poor aggregation. Therefore, impaired aggregation to collagen cannot solely explain the minimal inflammation after fetal wounding. PMID- 9200081 TI - The effects of tracheal occlusion and release on type II pneumocytes in fetal lambs. AB - Fetal tracheal occlusion (TO) has been shown to lead to lung hyperplasia in various animal models, and this procedure has already been carried out in human fetuses with congenital diaphragmatic hernia (CDH). However, the authors previously showed that TO caused a decrease in type II pneumocytes. PURPOSE: The aim of this study is to examine the effects of TO and release on type II pneumocytes. METHOD: To was carried out with a Swan Ganz or Fogarty catheter in fetal sheep at 116 to 118 days of gestation. TO was maintained for 2 weeks followed by deflation of the balloon for 1 week before delivery, in group 1; in group 2, TO was maintained for 19 days and released 2 days before delivery. Group 3 consisted of previously reported animals who had TO maintained until birth. Unoperated twins served as controls. All specimens were analyzed using the surfactant protein C (SP-C) mRNA as a specific marker for type II pneumocytes. We used Northern Blot and in situ hybridization techniques to quantify total SP-C and the density of type II cells. Electron microscopy (EM) was also used to evaluate and quantitate type II cells. RESULTS: TO resulted in significant lung growth in all groups. In situ hybridization and Northern Blot analysis showed that there was a complete recovery of type II cells in group 1 versus controls. Quantitative EM analysis confirmed these findings. In group 2 the number of type II cells was decreased but there was an increase in SP-C content per type II cell versus group 3. CONCLUSION: Lung growth after TO appears to occur at the expense of type II cell differentiation. This effect is reversible with the release of TO before birth in this animal model. PMID- 9200082 TI - A comparison of anal endosonography with electromyography and manometry in high and intermediate anorectal anomalies. AB - Anal endosonography, electromyography (EMG) of the external anal sphincter (EAS), and manometry of the internal anal sphincter (IAS) were performed in 15 patients with anorectal anomalies (10 with high and five with intermediate anomalies), ranging in age from 8 to 18 years. The anal endosonographic findings were compared with those for the EMG of the EAS and manometry of the IAS. An image including the hyperechoic band that corresponds to the EAS was obtained in all 15 patients. However, the distribution of EAS image was inadequate in high anomalies. In four patients who showed in Kelly score of 5 or 6, good visualization of the EAS was obtained in both anal endosonography and EMG. An Image including the hypoechoic band that corresponds to the IAS was obtained in five patients with high anomalies and in one with intermediate anomalies. Therefore, even in patients with anomalies, at these levels the IAS could be ultrasonically detected. However, only one of these six patients exhibited an anorectal reflex. These results indicate that, for the EAS, the findings of anal endosonography correspond well with those of EMG, but that for the IAS, they do not correspond with those of manometry. At the time of surgery for anorectal anomalies care should taken to preserve the IAS, which can be detected by anal endosonography even in patients with high or intermediate anomalies. PMID- 9200083 TI - The Pediatric Bowel Management Clinic: initial results of a multidisciplinary approach to functional constipation in children. AB - The multifactorial nature of functional constipation in children suggests that a multidisciplinary management approach may be effective. The authors tested this hypothesis in a newly created pediatric Bowel Management Clinic (BMC). Detailed data were collected prospectively on all patients seen in the clinic over the first 16 months. Both quantitative and qualitative analyses were performed to describe the index population and to demonstrate the impact of the intervention. Satisfaction with care in the clinic was measured using the Measure of Processes of Care tool, then compared with a normative sample. One hundred fourteen patients, all previously treated unsuccessfully for constipation, were referred to a team comprised of a physician, nurse practitioner, nurse educator, dietitian, and psychosocial nurse specialist. The mean age was 5.4 years with equal gender distribution. Between the first and last visits recorded, several variables including stool consistency and frequency, soiling frequency, abdominal pain, rectal pain, and rectal bleeding all showed statistically significant (P < .05) improvement. Qualitative data analysis showed the significant psychosocial impact of constipation on patients and their families. In the Measures of Processes of Care questionnaire, scores for the BMC were higher than normal on all scales except in provision of information. A multidisciplinary approach to functional constipation leads to both patient and parent satisfaction and significant short-term improvement. Further studies will examine the long-term impact of the clinic. PMID- 9200084 TI - The cecostomy button. AB - Percutaneous insertion of a cecostomy tube, performed under local anesthesia, to facilitate antegrade colonic cleansing, has been an invaluable advance in the management of fecal incontinence. However, the patient is left with a length of tubing (2 to 4 inches) protruding from the cecostomy site that has to be taped down to the abdominal wall. Available devices for insertion in place of the cecostomy tube are cumbersome and have a relatively high profile, projecting more than 1 cm from the surface of the abdominal wall. Worn under a swimsuit, they are clearly discernible. The inflated balloon within the cecum can occasionally break. Furthermore, in the individual with a relatively thick abdominal wall, such devices are too short to reach from the skin to the cecum. A new form of low profile trapdoor device has been developed that overcomes the above shortcomings of other available "buttons." It has been successfully used in a clinical setting in 49 patients. PMID- 9200085 TI - Anterior exposure of spinal deformities and tumors: a 20-year experience. AB - Data from 505 patients (1976 through 1995) who underwent anterior spinal exposure were retrospectively analyzed. There were 222 boys and 283 girls with a mean age of 14.5 years; 166 had thoracic exposure (T), 300 thoracoabdominal (TA), 44 retroperitoneal (R), and 7 transperitoneal (TP); 17 had repeat exposure (5 had initial exposure elsewhere); 70% had scoliosis, 25% kyphosis, 27% a neuromuscular disorder (NMD) and 6.7% a tumor. Average intensive-care-unit stay was 2.5 days, 6.2 days for NMD (P < .05); average ileus was 3.4 days, 4.1 days for NMD (P < .05); and average length of stay was 15.4 days for all patients, 19.3 days for NMD (P < .05). Mechanical ventilation over 96 hours was required in 31 patients, 66% had an NMD (P < .05). The morbidity rate was 9.8%, 10.1% for NMD; the morbidity rate was zero for tumor and repeat exposures. Mortality was zero. Over half of the vessel injuries (57%) and the urinary tract infections (60%) occurred in NMD patients. Differences between the 1976 through 1985 period and the 1986 through 1995 period were a shorter length of stay and a majority of one-stage combined exposures in the latter period. The authors conclude that anterior exposure of spinal deformities is well tolerated by most pediatric patients, and that this technique is easily adaptable to the resection of retroperitoneal and thoracolumbar tumors. PMID- 9200086 TI - Surgical pancreatic complications induced by L-asparaginase. AB - Pancreatitis has been noted to be a potential complication in 2% to 16% of patients undergoing treatment with L-asparaginase for a variety of pediatric neoplasms, but rarely has surgical intervention been necessary. The authors present two fulminant cases of L-asparaginase-induced pancreatitis and review the current literature. The first patient is a 15-year-old boy who underwent induction chemotherapy with L-asparaginase for non-Hodgkin's lymphoma with bone marrow involvement. He presented with diffuse patchy necrosis of the pancreas as well as a large infected pancreatic pseudocyst. He subsequently required operative debridement of the pancreas and external drainage of the pseudocyst. He is currently doing well. The second patient is a 5-year-old boy who was treated with L-asparaginase for a diagnosis of acute lymphocytic leukemia. Within 3 weeks of initiation of therapy, fulminant pancreatitis developed, which progressed to multisystem organ failure. Computed tomography scan demonstrated extensive pancreatic necrosis involving 90% of the gland. He underwent surgical debridement of his necrotic pancreas and wide drainage of the lesser sac. Postoperatively he improved but subsequently multiple complications developed including erosion of his gastroduodenal artery with significant intraabdominal bleeding, which was controlled with angiographic embolization. Subsequently erosion of his endotracheal tube into the innominate vein developed, and he died. L-asparaginase induced pancreatitis has been described after therapy for various pediatric neoplasms, and the reported cases have usually been self-limiting. However, our cases demonstrate potentially fatal sequelae of this complication and mandate early diagnosis with appropriate surgical intervention in this setting. PMID- 9200087 TI - Scrotoschisis associated with contralateral meconium periorchitis. AB - Scrotoschisis, a congenital defect of the scrotal wall associated with extracorporeal testicular ectopy, has been previously reported only twice. Meconium periorchitis is another rare scrotal anomaly indicative of an antenatally healed gastrointestinal perforation. The authors present a third case of scrotoschisis and the first associated with meconium periorchitis. Several hours after birth of an otherwise-normal term baby boy, a scrotal exploration was performed with orchidopexy and primary closure of the scrotal wall defect. At 4 months of age the baby underwent a contralateral inguino-scrotal exploration with excision of a paratesticular mass of calcified meconium. The role of a normally developed scrotum in testicular descent and causes of calcified scrotal masses in infants are discussed. PMID- 9200089 TI - Empyema thoracis in children: a 26-year review of the Montreal Children's Hospital experience. AB - The appropriate management of pediatric empyema thoracis remains controversial. The authors reviewed 47 cases of empyema thoracis over a 26-year period. The management of empyema included initial diagnostic thoracentesis and classification as acute, fibropurulent, or chronic. If the empyema was "acute," therapeutic tap, tube thoracostomy, or no surgical intervention was performed. "Fibropurulent" empyemas were uniformly treated with tube thoracostomy. The lung was decorticated when the empyema was encased by a thick peel, had recurred and was multiloculated, was refractory and the patient remained clinically unwell, or had occurred as a complication of previous thoracotomy. All patients with acute empyemas responded to antibiotics irrespective of drainage (average duration of fever, 17 days; average stay in hospital, 27 days). Of the fibropurulent empyemas in our review, complete drainage was attained in seven of 39 (18%), and decortication was not required in any empyema that was completely drained. Loculations persisted in 25 of 39 (64%) after tube thoracostomy but nonetheless resolved. The remaining seven of 39 (18%) with persistent loculations required formal decortication. Of the patients with fibropurulent empyemas that responded to tube thoracostomy, the average duration of fever was 13 days and hospitalization, 23 days. Of those requiring decortication the average duration of fever was 24 days and hospitalization, 40 days. These results will allow a baseline for comparison of new strategies (fibrinolytics and early thoracoscopy) that may reduce days of fever, hospitalization, and risk of formal decortication. PMID- 9200088 TI - Anal reeducation for postoperative fecal incontinence in congenital diseases of the rectum and anus. AB - From October 1993 to March 1996, 14 patients with anorectal disease were referred to an anal reeducation clinic. Initial evaluation allowed the authors to identify three classes of defects: lack of proprioception in the sphincters, use of synergistic muscles (gluteal) to compensate for anal dysfunction, and inversion of command by contraction, rather than relaxation, of abdominal muscles. Patients were treated by electrostimulation through an anal probe as well as biofeedback therapy coupled with home exercises. This therapy resulted in rapid correction of the abnormal motor commands and erroneous use of accessory muscles. All patients became able to isolate their continence muscles with success, with documented increase in strength, rapidity of response, and duration of contraction. The mean Kelly score went from 1.46 (range, 0 to 4.5) to 3.07 (range, 0.5 to 5.5). This physiological improvement also increased patient motivation and discipline toward continence and subsequently their quality of life. PMID- 9200090 TI - Intestinal vascular anomalies in children. AB - Vascular anomalies are an uncommon cause of gastrointestinal bleeding in childhood. Confusing nomenclature has made objective comparisons of published cases difficult and has interfered with an established consensus regarding diagnosis and therapeutic modalities. The purpose of this study was to clarify the situation by reviewing the records of all children who had intestinal vascular anomalies who were referred to our institution from 1975 to 1995. Thirteen lesions were identified in nine children (five boys and four girls). The median age at clinical onset was 8 years. Only two patients presented with a complex syndrome (Klippel-Trenaunay, 1; Osler-Rendu-Weber, 1). Diagnosis, location, and extension of these anomalies was only possible by angiography, which indicated that seven patients had isolated venous malformations and two had arteriovenous malformations. Because the lesions did not involve the serosa, intraoperative localization was a major problem. The main findings were a few slightly dilated mesenteric veins. Treatment was conservative in four children and surgical in five. Pathological findings on resected bowel demonstrated dilated and abnormal veins in the mucosa and submucosa. Selective angiography should not be delayed in patients with gastrointestinal bleeding if results of all other investigations are negative. Because these lesions are rarely recognizable on operative inspection, precise preoperative angiographic localization of intestinal vascular anomalies is essential to allow for a safe and limited resection of the involved bowel segment. Based on a better understanding of the natural history of these lesions, a classification of vascular anomalies of intestines in children is proposed. PMID- 9200091 TI - Proximity injury by the ultrasonically activated scalpel during dissection. AB - The ultrasonically activated scalpel is a high-frequency oscillating instrument that is reported to have a decreased dispersion of energy to surrounding tissues during use. To determine if this effect is beneficial and safe to surrounding tissue, it was used on anesthetized adolescent swine to dissect the portal vein from the pancreas, the renal artery and vein from the renal hilum, the ureter from the retroperitoneum, the aorta from the inferior vena cava and the common bile duct from surrounding tissue. Three-second contact to intestine and nerve roots was also performed. Wedge biopsy specimens of liver and spleen were performed. Dissection technique used was as described by the company. Structures were dissected with electrocautery using similar techniques for comparison. Tissues were harvested and placed in formalin for histological analysis. Dissection with the ultrasonically activated scalpel was simple, achieved excellent hemostasis, and did not appear to damage adjacent tissue. Microscopic analysis showed adventicial and media injury to vascular structures. The ureter and common bile duct demonstrated marked injury with regions of transmural coagulation. Nerve and small bowel did not appear to have much injury from the 3 second contact with the instrument. This study indicated that although the ultrasonically activated scalpel can ease dissection with good hemostasis, care must be taken to avoid injury to adjacent structures. Although its lateral energy dispersion may be less than that of cautery, it can still cause transmural necrosis to major structures. PMID- 9200092 TI - Diagnosis and management of duodenal injuries in children. AB - Traumatic duodenal perforations in children pose a diagnostic and therapeutic challenge. To identify specific diagnostic criteria and define an optimal therapeutic protocol, we reviewed all duodenal injuries treated at our institution in the past 10 years. There were 14 hematomas and 13 perforations. The diagnosis was confirmed by computed tomography (CT), ultrasound scan (US), upper gastrointestinal contrast studies (UGI), or at laparotomy. The clinical findings and CT findings of the two groups were compared. Children with suspected duodenal hematomas were treated expectantly, and children with duodenal perforations were treated surgically. Twenty-five associated injuries (10 pancreatic) occurred in 19 children. Children with perforations had higher injury severity scores (ISS) (25 v 9), but the two groups could not be differentiated based on presenting signs, symptoms, or laboratory findings. CT findings of retroperitoneal air or contrast were seen in 9 of 9 perforations and in 0 of 10 hematomas. CT findings of intraabdominal or retroperitoneal fluid, mesenteric enhancement, and thickened duodenal wall did not differentiate the two groups. Duodenojejunostomy was performed in one patient, and primary repair was performed in 11 children who had perforation. In five children, duodenostomy tube drainage with feeding jejunostomy or gastrojejunostomy were added. Complications occurred in three of four children in the first 5 years of the study and in two of nine children in the last 5 years. The decreased morbidity rate correlated with reduced time to definitive therapy (28 v 7.8 hours). Duodenal fistulae resulted in three of seven children treated without duodenostomy tube drainage and zero of five treated with drainage. Enteral feeds resumed faster (average, 12 v 27 days) if repair of perforation was combined with feeding jejunostomy or pyloric exclusion and gastrojejunostomy. Children with duodenal hematoma resumed eating an average of 16 days after injury. Only one child required surgery for persistent obstruction. The findings of retroperitoneal air and contrast extravasation on CT accurately distinguish duodenal perforation from hematoma. Conservative management of hematoma is safe and effective. Primary repair of perforation with duodenal drainage results in fewer postoperative complications, and gastrojejunostomy or feeding jejunostomy shorten the time to resumption of feeds. PMID- 9200093 TI - The uncut Collis-Nissen fundoplication: results for 79 consecutively treated high risk children. AB - PURPOSE: The Nissen fundoplication fails to control gastroesophageal reflux (GER) in up to 25% of children with neurological impairment or chronic lung disease. The uncut Collis modification lengthens the intraabdominal esophagus, improving the antireflux function without opening the stomach. This study reviews the results of the uncut Collis-Nissen fundoplication in a pediatric series. METHODS: Seventy-nine children had an uncut Collis-Nissen fundoplication performed over a 5-year period. The median age was 1.4 years. Associated problems included neurological impairment (77%), chronic lung disease (38%), and esophageal atresia (3%). Surgery was undertaken only in children with objective documentation of pathological GER, who had GER complications unresponsive to medical treatment. The usual complications that led to surgery were pulmonary (73%), esophagitis (67%), or failure to thrive (35%). Liquid gastric emptying was assessed routinely preoperatively, and was delayed in 42% patients who then had concomitant pyloroplasty. RESULTS: GER was controlled in 97% of patients after a median follow-up of 1.8 years. All children with recurrent symptoms were restudied, and only two children had documented recurrent GER. One of these required a repeat fundoplication. Thirty-three percent were on promotility medication for feeding difficulties, gagging, or retching. There were postoperative complications in 26% (minor 23%, major 3%) and one postoperative mortality. Eleven late deaths were unrelated to surgery or GER. CONCLUSION: The uncut Collis-Nissen fundoplication provides excellent control of GER in children and is associated with acceptable morbidity and low mortality. It should be particularly considered in children with neurological impairment or chronic lung disease. PMID- 9200094 TI - Assessment of the postoperative visit after routine inguinal hernia repair: a prospective randomized trial. AB - Treatment of pediatric patients undergoing routine inguinal hernia repair usually includes a postoperative clinic visit. We prospectively assessed the necessity for the traditional postoperative visit. One hundred patients undergoing a routine inguinal hernia repair were randomly selected to receive either a follow up visit at 4 weeks or a detailed instruction sheet and no follow-up visit. Parents were given a telephone questionnaire to determine overall satisfaction with their child's care and the usefulness of the follow-up visit or instruction sheet. Forty-seven of 50 parents of patients randomly assigned to a follow-up clinic visit (FU) and all 50 parents in the no follow-up group (NFU) completed the questionnaire. Sixty-eight percent of the FU group found the follow-up visit "helpful" and 59% found it "necessary." Fifty-six percent would have been satisfied with a telephone call instead of a visit. In the NFU group only 4% thought a follow-up visit would have been "helpful" and 4% thought a visit was "necessary." Ninety-six percent found the postoperative instruction sheet "helpful." There was no difference between groups in overall satisfaction with the care received as assessed on a 5-point scale (4.7 FU group v 4.7 NFU group). Accurate postoperative instruction and open access to follow-up when required is as effective as the traditional postoperative clinic visit for patients who have undergone routine inguinal hernia repair. PMID- 9200095 TI - Laparoscopic pull-through procedures using the harmonic scalpel in infants and children with Hirschsprung's disease. AB - Hirschsprung's disease in infants has routinely been treated by a three- or four stage process requiring a rectal biopsy, diverting colostomy, pull-through procedure, and then colostomy takedown. This algorithm requires multiple hospitalizations and surgeries over several months. The authors have adopted a laparoscopic approach that allows the surgery to be performed in one stage with a marked decrease in morbidity and hospital stay. From March 1995 to May 1996, 15 infants and children, ages 7 days to 8 years and weighing 2.3 kg to 40 kg, underwent laparoscopic pull-through procedures. Eleven underwent primary pull through, while four underwent a previous diverting colostomy. The laparoscopic portion of the pull-through was performed using three or four ports, size 3.5 mm or 5 mm and an ultrasonic dissector. The final submucosal dissection was performed transrectally starting 1 cm above the pectinate line. The rectal anastomosis was hand sewn, and no patient was left with a diverting colostomy. Operative time averaged 2 hours and 50 minutes. Average time to feeds was 1.3 days and the average days to discharge was 3.4. There was one intraoperative pathology misdiagnosis and one patient with an anastomotic stricture. All patients are excreting stools spontaneously at least daily and there have been no episodes of colitis. This preliminary report shows that the one-stage laparoscopic pull-through is safe and effective. PMID- 9200096 TI - Morphological changes in the enteric nervous system of the transplanted fetal rat intestine. AB - In this study, enteric nervous system (ENS) of the fetal intestinal grafts was examined histopathologically. Forty-four rat fetal small intestines were transplanted syngenetically into the subcutaneous region of adult rats without vascular anastomosis. Thirty-two grafts survived. They were removed 2, 4, 6, and 8 weeks after transplantation and examined using (1) H&E staining, (2) AChE and NADPH-diaphorase histochemistry, and (3) protein gene product 9.5, S-100 protein, glial fibrillary acidic protein, tyrosine hydroxylase, nerve growth factor receptor, calcitonin gene-related peptide, neuropeptide Y, vasoactive intestinal peptide, somatostatin, and substance P immunohistochemistry. The grafts were compared with the intestines of 2-, 4-, 6- and 8-week-old control rats. ENS of the grafts was different from the controls as follows: (1) tyrosine hydroxylase and neuropeptide Y were markedly reduced but present, suggesting that the extrinsic innervation was present; (2) nitric oxide-producing neurons were well preserved in grafts; (3) hyperganglionosis in the myenteric plexus was seen in 6- and 8-week grafts; (4) AChE activity was increased in the circular muscle and in the lamina propria, (5) S-100 was increased in the lamina propria in 6- and 8 week grafts, (6) calcitonin gene-related peptide was increased in 6- and 8-week grafts, (7) nerve fibers in the muscle layers ran irregularly and disorderly, and (8) hypertrophy of smooth muscle layers. Our data show that although extrinsic as well as intrinsic innervation is present in the fetal intestinal grafts, there is hyperinnervation of the intrinsic nervous system and reduced innervation of the extrinsic ENS. These morphological changes in the ENS of the fetal intestinal grafts may result in motility dysfunction. PMID- 9200097 TI - Intrahepatic biliary-enteric bypass for complete extrahepatic biliary obstruction in children. AB - Intrahepatic biliary-enteric anastomosis is rarely practiced in pediatric surgery. The authors report on two children who have been successfully treated using this method. First described by Longmire and Sandford in 1948, intrahepatic biliary-enteric anastomosis is possible in children. This procedure is only indicated as a last recourse for circumstances in which the extrahepatic biliary tree is completely obstructed. PMID- 9200098 TI - Pentoxifylline inhibits overflow and reduces intestinal reperfusion injury. AB - The aim of this study was to determine the effects of pentoxifylline (Ptx) in reperfusion injury of the small bowel as a leukocyte stabilizer, free radical scavenger, and microcirculatory regulator. Ninety-six male Sprague-Dawley rats were used to determine the biochemical, histopathologic and blood flow changes of the reperfused small intestines after 30 minutes of a warm ischemic insult. Animals were divided into six groups: Sham (S), sham plus Ptx (SP), ischemia (I), ischemia plus Ptx (IP), reperfusion (R), and reperfusion plus Ptx (RP). Pentoxifylline was administered intraperitoneally at a dose of 50 mg/kg 15 minutes before ischemia. The superior mesenteric artery (SMA) was occluded distal to the right colic artery and collateral arcades were ligated as described by Megison. Sixty of the 96 rats (n = 10) were used to determine histopathologic changes, malondialdehyde (MDA), and myeloperoxidase (MPO) levels in tissue. Mucosal lesions were graded on a scale from 0 to 5 as described by Chiu. MDA and MPO levels of the intestinal mucosa were assayed to reflect the free radical formation and neutrophil sequestration, respectively. Thirty-six rats (n = 6) were used to measure blood flow changes of the intestine using 133Xe clearance technique. All data were presented as the mean values plus or minus the standard error of the means (means +/- sem). Although in the R group, mucosal injury score, blood flow, MPO, and MDA levels were higher significantly from the other groups (P < .05), in the RP group blood flow, MPO, and MDA levels were significantly decreased to the basal values (P < .05). Mucosal injury score of the RP group were lower than the reperfusion group but higher than the normal (P < .05). The authors conclude that pentoxifylline pretreatment before reperfusion stabilizes blood flow, decreases MPO and MDA levels to the normal, and attenuates but not completely prevents mucosal damage. PMID- 9200099 TI - Childhood adrenocortical tumors: case series and reevaluation of prognosis--a 24 year experience. AB - Adrenocortical neoplasms are rare in childhood and adolescence. The prognostic significance of tumor size, weight, and histological grade are still very much unclear. Eleven patients, (3 boys, 8 girls), with a median presentation age of 7 years (range, 0.8 to 16 years) were identified. Six presented with virilizing symptoms, two with cushingoid symptoms, one with both, and two others had nonspecific symptoms. The interval between onset of symptoms and diagnosis was an average of 18 months (median, 8 months). Hormonal profile correlated well with clinical presentation in nine patients. Two patients with nonspecific symptoms had an aldosterone-producing lesion and an androgen-secreting tumor. Ten patients underwent complete surgical excision, with one intraoperative spillage. Median tumor weight was 94.5 g (range, 4 to 750 g). Three lesions were less than 5 cm in maximal width, six were between 5 and 10 cm, and two were greater than 10 cm. Two tumors had capsular or vascular invasion. Three patients received chemotherapy: one who had inoperable metastatic disease, and two based on clinical and histopathologic findings. Ten patients are doing well, without evidence of recurrent disease with a median follow-up of 3 years (range, 9 months to 15 years), eight patients have been followed up for more than 2 years. The medically treated patient who had metastatic disease died 3 years after diagnosis. A review of the pediatric literature, in some cases, indicates that larger tumors have a worse prognosis, while other investigators claim histological grade is more important. The authors' results do not support these conclusions, but rather suggest that in the pediatric population, when excision is complete, guarded optimism is warranted even with tumors larger than 5 cm. Addendum: Since submission of the manuscript, patient 4 has been operated on twice for local recurrences 13 and 16 months after the initial surgery. She was the only patient in the series to have an intraoperative capsular tear. All other surgical patients remain free of disease. PMID- 9200100 TI - Botulinum toxin use in pediatric esophageal achalasia: a case report. AB - Esophageal achalasia (EA) has been historically treated by esophageal dilatation or myotomy with or without fundoplication. Botulinum toxin (Botox-Allergan) use in pediatric EA has not been previously described. The authors' objective was to observe the efficacy of botulinum toxin injection into the lower esophageal sphincter (LES) for EA. An 11-year-old boy presented with a 9-month history of frequent pneumonia, productive cough, and a 1-year history of chest discomfort and odynophagia. Chest radiograph showed changes compatible with aspiration. Upper gastrointestinal (UGI) series showed typical narrowing of the LES, and 24 hour pH study showed no reflux. Esophageal manometry showed classic findings of achalasia. An upper gastrointestinal endoscopy was performed showing a huge volume of retained food. A direct four-quadrant injection was performed with a total of 100 U of botulinum toxin into the LES. UGI series showed improvement in esophageal emptying. Esophageal manometry showed impressive improvement in LES pressure (preinjection, 44.1 mm Hg to postinjection mean of 16.6 mm Hg), percent relaxation (preinjection, 30% to postinjection, 58.8%), and duration of relaxation (preinjection, 1.9 seconds to postinjection, 11 seconds). The patient has not had any further respiratory symptoms, chest pain, or odynophagia in 8 months of follow-up. Botulinum toxin injection is simple and effective for EA and merits its study in a prospective manner in the pediatric population. PMID- 9200101 TI - Management of impalpable testes: indications for abdominal exploration. AB - Various approaches to the management of the impalpable testis in cases of cryptorchidism have been advocated. The authors' experience over the past 13 years was reviewed to try to determine an optimal approach. Of 1,305 patients with undescended testicles seen between February 1982 and December 1995, 157 boys (12.03%) had impalpable testes with 17 having bilateral impalpable testes for a total of 174 impalpable testes. A hernia sac was present in 155 impalpable testes with a testicle present in all cases. No hernia sac was found in 19 impalpable testes, five of which had no testicle present. This was confirmed by either open exploration or laparoscopy. One hundred forty-eight boys underwent groin exploration as initial treatment, 13 of these had bilateral impalpable testes. In addition to the five absent testicles with no hernia sac, one patient with a hernia sac and no testicle evident benefited from subsequent laparoscopy to identify an intraabdominal testicle. All other patients underwent routine orchidopexy or orchidectomy (one case with grossly malformed testicle). Nine boys underwent laparoscopy as initial treatment, four of these had bilateral impalpable testes. Two abnormal testicles were found and removed. Groin exploration and subsequent orchidopexy was definitive treatment in all other cases. The association of a hernia sac with an impalpable undescended testicle is very significant (P < .00001 Fisher's Exact test). The absence of a sac therefore may reflect an alternate diagnosis. When no sac is found with a testicle in the groin, this may represent an ectopic testicle. When no sac is found with no testicle, this may represent a vanishing testicle. From this experience the authors conclude that groin exploration should be the initial approach to impalpable testes. The presence of a hernia sac with an absent testicle demands further exploration; the absence of a hernia sac with an absent testicle suggests a vanishing testicle and may need no further exploration. PMID- 9200102 TI - An unusual variant of rectal atresia with rectovestibular fistula. AB - Isolated rectal atresia in the girl with a normal anal canal is extremely rare, and its association with a fistula has not been reported in the literature. A 6 year-old girl who had a unique combination of rectal atresia and rectovestibular fistula was treated successfully by a posterior sagittal approach. PMID- 9200103 TI - Gastrointestinal myoelectric activity in a child with gastroschisis and ileal atresia. AB - Gastroschisis is frequently associated with intestinal atresia and alterations in gastrointestinal function. The authors studied gastric and small bowel myoelectric activity in a child who had a complex course and prolonged inability to tolerate oral intake after staged repair of gastroschisis and an associated ileal atresia. The child remained unable to tolerate oral intake after repair of the atresia and was reexplored 3 months later to rule out a partial small bowel obstruction, with simultaneous placement of serosal electrodes on the stomach and proximal small bowel. Persistent gastric dysrhythmias were observed postoperatively, and the child was unable to tolerate gastrostomy tube feedings. Abnormalities were also seen in small bowel motility, including retrograde propagation of activity fronts of the migrating myoelectric complex. However, the intestine converted to a fed myoelectric pattern with tube feedings, and the child was subsequently able to tolerate feedings via a tube placed directly into the small bowel. The authors conclude that myoelectric recordings via implanted electrodes are safe and feasible in children, and may give information regarding underlying motility alterations. The ultimate clinical role of myoelectric recordings in treating children with suspected motility disorders will require further study. PMID- 9200104 TI - Acquired gastric outlet obstruction during infancy and childhood: a report of five unusual cases. AB - Infantile hypertrophic pyloric stenosis is a common, specific, and well understood entity in the pediatric age group. Prepyloric webs and diaphragms are the other rare causes of gastric outlet obstruction followed by still rarer causes, eg, pyloric stenosis secondary to acid peptic disease. The present report describes a newer, nonspecific, clinical condition of pyloric obstruction in five patients of varying age who responded very well to Henieke-Mikulicz' pyloroplasty. The authors have suggested some modifications in the classification of gastric outlet obstruction in children to include these patients. PMID- 9200105 TI - Early history of the therapy of Hirschsprung's disease: facts and personal observations over 50 years. PMID- 9200106 TI - Functional bowel stasis in the dilated segment of proximal bowel among newborns presenting with duodenal or jejunal atresia. PMID- 9200108 TI - Is delayed surgery really better for congenital diaphargmatic hernia? PMID- 9200107 TI - Upper esophageal stenosis: two case reports. PMID- 9200109 TI - Immediate reconstruction for penile agenesis. PMID- 9200110 TI - What is your diagnosis? Eosinophilic keratoconjunctivitis. PMID- 9200111 TI - Lipomatous infiltration of the canine salivary gland. AB - Benign connective tumours of the canine salivary glands are rare. This report describes lipomatous infiltration of parotid or submandibular salivary glands in seven dogs in which the glands were enlarged as a result of infiltration by fat cells; they appeared to have been successfully treated by local excision. The precise cause of the lipomatous infiltration in the dogs is unclear but different causes of similar lesions in humans are discussed. PMID- 9200112 TI - Catheter drainage of pleural fluid collections and pneumothorax. AB - A technique for virtually atraumatic placement of small size chest catheters for suction drainage of pleural effusions and pneumothorax in the dog and cat is described. Thirty-nine dogs and two cats were treated for pyothorax (10 cases), hydrothorax (eight), chylothorax (three), haemothorax (three), haemothorax/ pneumothorax (three) and pneumothorax (14). In all 41 cases, thin or viscous fluid and/or air were efficiently drained. The mean period of drainage was four days (range, 0.5 to 18 days). The average amount of fluid removed from each patient in 24 hours was 530 ml in pyothorax cases (range, 140 to 1100 ml) and 1300 ml in the other cases (range, 20 to 5000 ml). In 40 cases there were no complications related to the procedure. One dog with severe pleural adhesions was euthanased because of lung perforation and pneumothorax secondary to misplacement of the catheter. PMID- 9200113 TI - Potentiating effect of EDTA-Tris on the activity of antibiotics against resistant bacteria associated with otitis, dermatitis and cystitis. AB - Possible synergistic effects of the combination of EDTA-tromethamine (EDTA-Tris) and three antimicrobial agents (cephaloridine, kanendomycin and enrofloxacin) against resistant Gram-positive and Gram-negative bacteria are reported. Bacteria were isolated from eight cases of chronic otitis externa, five cases of chronic dermatitis and four cases of recurrent cystitis in dogs which had previously been treated with one of the three antibiotics without success. Animals exposed to EDTA-tromethamine plus the antibiotic recovered completely within 10 days, and were controlled clinically and bacteriologically for 180 days. Local irrigation with EDTA-tromethamine solution was well tolerated and no side effects were recorded. PMID- 9200115 TI - Surgical management of subvalvular aortic stenosis and mitral dysplasia in a golden retriever. AB - A 12-month-old neutered male golden retriever was presented with a history of lethargy and exercise intolerance. Clinical examination, electrocardiography, radiography and echocardiography supported a diagnosis of fixed subvalvular aortic stenosis with a Doppler pressure gradient of 77.5 mmHg. Surgical inspection also revealed gross structural abnormalities of the mitral valve consistent with mitral dysplasia. Intervention consisted of resection of the dysplastic mitral valve and the subvalvular aortic stenosis. The mitral valve was replaced with a bioprosthetic valve. Total cardiopulmonary bypass time was 65 minutes and aortic cross-clamp time was 55 minutes. A full recovery was made and 11 months postoperatively the aortic transvalvular gradient was 30 mmHg. At the time of writing, 12 months after surgery, the dog was clinically normal and requires no medication. PMID- 9200114 TI - Superficial necrolytic dermatitis and a pancreatic endocrine tumour in a dog. AB - A 13-year-old dog was referred for a severe dermatological problem of 12 months duration. Skin biopsy results were compatible with superficial necrolytic dermatitis. The only laboratory abnormalities were hyperglycaemia and hyperglucagonaemia. These findings suggested a pancreatic endocrine tumour in association with superficial necrolytic dermatitis. Abdominal ultrasound examination was unremarkable. The dog was euthanased due to the lack of clinical improvement following symptomatic therapy. Postmortem examination revealed a pancreatic endocrine tumour with liver metastases. Pancreatic endocrine tumour cells were immunoreactive for glucagon, insulin and islet amyloid polypeptide. PMID- 9200116 TI - Bilateral pigmented villonodular synovitis in a dog. AB - Villonodular synovitis is an extremely rare condition of the synovial membrane in the dog. A 10-year-old, neutered crossbreed was presented with bilateral, progressive hindlimb lameness. Periarticular swelling was noted in both stifle joints. No craniocaudal instability was noted. Radiographs showed massive intra articular soft tissue proliferation in both joints, with no bony involvement. Arthrocentesis was unsuccessful. Exploratory arthrotomy of the left stifle revealed a greatly thickened, florid, proliferative synovial membrane. An incisional biopsy was carried out and the histopathological diagnosis was chronic active villonodular synovitis. A radical synovectomy was carried out in the right stifle joint 10 days later. Corticosteroid treatment was initiated 10 days after the second surgery and continued for six weeks, with a continuous clinical improvement. Eighteen months after discontinuation of the steroid therapy, the owners reported no recurrence of clinical signs although a mild stiffness was still present. PMID- 9200117 TI - Use of a 0.25 per cent fipronil pump spray formulation to treat canine cheyletiellosis. AB - Two outbreaks of cheyletiellosis are described. In one of the outbreaks, a human member of the household was also affected. A 0.25 per cent fipronil pump spray formulation was applied to the affected and in-contact animals. A permethrin spray was used in an attempt to eliminate environmental contamination. One month later, the affected animals were re-examined. No evidence of Cheyletiella mites could be found. A second application of fipronil was undertaken. No further outbreaks were reported during an eight month follow-up. PMID- 9200118 TI - Osteosarcoma following total hip arthroplasty in a dog. AB - Osteosarcoma involving the distal right femur was diagnosed in a nine-year-old female neutered Rottweiler seven years after total hip arthroplasty had been performed on that limb. The findings were consistent with a primary bone tumour and pathological fracture of the right femoral condyle with loosening of the orthopaedic implant and fracture of the polymethylmethacrylate at the distal aspect of the femoral component. Possible hypotheses to explain the association of osteosarcoma with total hip arthroplasty suggest that the neoplastic process was the result of some derangement of host tissue and the healing process or that the implants or their by-products were carcinogenic. Given the large number of total hip arthroplasties that are routinely performed in dogs, the development of a malignant lesion appears to be an extraordinary complication and may be completely coincidental. PMID- 9200119 TI - Zoonotic diseases: putting the risks in perspective. PMID- 9200120 TI - The genomic organization of guide RNA genes in kinetoplastid protozoa: several conundrums and their solutions. AB - The guide RNA (gRNA) paradigm states that the uridine (U) insertion/deletion type of RNA editing is mediated by short 3' uridylylated gRNAs that are complementary to specific blocks of mature edited sequence. These gRNAs contain the edited sequence information in the form of guiding purine residues that can base pair with the inserted U's and do not base pair with encoded U's that are to be deleted. The minicircle gRNA genes in trypanosomatids are localized at specific sites within the variable region, with the number and the precise localization of genes also being species-specific. The total number of minicircle sequence classes and thereby minicircle-encoded gRNAs varies greatly between species and even between different strains of the same species, with the greatest number being in the trypanosome species. Several conundrums which appeared to raise problems for the gRNA paradigm arose during comparative analysis of minicircle gRNA gene organization. The solution of these conundrums has led to a better understanding of the function and evolution of this RNA modification phenomenon. PMID- 9200121 TI - cDNA cloning of galectins from third stage larvae of the parasitic nematode Teladorsagia circumcincta. AB - A monoclonal antibody raised to a Teladorsagia circumcincta 31-33 kDa doublet antigen was used to immunoscreen a T. circumcincta cDNA expression library. Sheep antibodies eluted from the proteins expressed by two clones immunopositive with the monoclonal antibody specifically recognised the doublet antigen on Western blots of third stage larval extract, confirming that these clones coded for the antigen. Database searches revealed high levels of similarity with beta galactoside-binding lectin-like proteins (Ga1BPs or galectins) from Caenorhabditis elegans and Onchocerca volvulus. By analogy with these sequences, both T. circumcincta cDNA clones contain the full-length protein coding region. The native doublet proteins could be preferentially extracted from homogenates of third stage larvae with lactose and could be affinity purified on an asialofetuin column, confirming the identity of these bands as galectins. Reverse transcriptase-polymerase chain reaction amplification using a primer based on the C. elegans Spliced Leader SL1 sequence showed that the corresponding T. circumcincta mRNAs are also trans-spliced at their 5' ends. While there are considerable nucleotide differences between the two clones, the majority are located in the non-coding regions. Within the coding region there are 87 nucleotide differences but only three of these result in amino acid substitutions. PMID- 9200122 TI - Replication, expression and segregation of plasmid-borne DNA in genetically transformed malaria parasites. AB - To fully exploit the transfection technology developed for Plasmodium we investigated the features of replication, expression and segregation of an episomally maintained DNA construct during a sexual blood stage development in genetically transformed parasites of P. berghei. Using DNA in situ hybridisation techniques we were able to show that the introduced DNA construct is located in the nucleus of the parasite and is not segregating uniformly during schizogony. Replication of the construct mainly takes place between 16 and 24 h after invasion of the merozoites, coinciding with chromosomal replication. Furthermore the plasmid-borne DHFR/TS gene is constitutively transcribed throughout the asexual blood stage development. Hence the DHFR/TS promoter would appear to be a useful tool in the study of (over)expression of introduced genes and performing complementation studies in transfected parasites during the complete a sexual blood stage development of P. berghei. PMID- 9200123 TI - Manipulation of the vsg co-transposed region increases expression-site switching in Trypanosoma brucei. AB - Disruption of a region of DNA in Trypanosoma brucei immediately upstream of the expressed telomere-proximal variant surface glycoprotein gene (vsg), known as the co-transposed region (CTR), can cause a dramatic increase in the rate at which the active expression site (ES) is switched off and a new ES is switched on. Deletion of most of the CTR in two ESs caused a greater than 100-fold increase in the rate of ES switching, to about 1.3 x 10(-4) per generation. A more dramatic effect was observed when the entire CTR and the 5' coding region of the expressed vsg221 were deleted. In this case a new ES was activated within a few cell divisions. This switch also occurred in cell lines where a second vsg had been inserted into the ES, prior to CTR deletion. These cell lines, which stably co expressed the inserted and endogenous Vsgs, in equal amounts, did not differ from the wild-type in growth rate or switching frequency, suggesting that simultaneous expression of two Vsgs has no intrinsic effect. CTR deletion did not disturb the inserted vsg117. We tentatively conclude that it was not the disruption of the vsg221 in itself that destabilized the ES. All of the observed switches occurred without additional detectable DNA rearrangements in the switched ES. Deletion of the 70-bp repeats and/or a vsg pseudogene upstream of the CTR did not affect ES stability. Several speculative interpretations of these observation are offered, the most intriguing of which is that the CTR plays some role in modulating chromatin conformation at an ES. PMID- 9200124 TI - Identification of hemoglobin degradation products in Plasmodium falciparum. AB - Malaria parasites break down human hemoglobin to its constituent amino acids by cysteine and aspartic proteinases. However, no one has previously been able to identify hemoglobin cleavage products in intact parasites. When isolated parasites were subjected to non-denaturing polyacrylamide gels electrophoresis, a unique protein band was found which contains heme and reacts with anti-human hemoglobin antibodies. This protein does not appear to represent oxidized or glycosylated hemoglobin, and is present in isolated parasites but not in the cytosol of infected or uninfected erythrocytes. When this band was eluted and subjected to SDS polyacrylamide gel electrophoresis, three bands were seen on Western blots. The proteins in these bands contain proteins with the N-terminal sequences of alpha- and beta-globin chains but molecular masses of only 13.2-13.4 kDa. These data suggest that hemoglobin alpha- and beta-chains are initially cleaved within the parasite phagolysosome to release peptides of 15-17 and 23-25 amino acids from the C-termini of alpha- and beta-globin chains, respectively. Production of the hemoglobin breakdown products was inhibited by E-64, a cysteine proteinase inhibitor, suggesting the involvement of a cysteine proteinase in an early step of hemoglobin degradation. PMID- 9200125 TI - Molecular cloning and characterization of two iron superoxide dismutase cDNAs from Trypanosoma cruzi. AB - Two cDNAs (FeSODA and FeSODB cDNAs) corresponding to superoxide dismutase (1.15.1.1., SOD) were isolated from a Trypanosoma cruzi cDNA library. Comparison of the deduced amino acid sequences with previously reported SOD protein sequences revealed that the T. cruzi open reading frames had considerable homology with FeSODs. The coding region of the T. cruzi FeSODB cDNA has been expressed in fusion with glutathione-S-transferase using an Escherichia coli mutant QC779, lacking both MnSOD and FeSOD genes (sodA sodB). Staining of native polyacrylamide gels for SOD activity of T cruzi crude lysate and the recombinant SOD suggests that this protein is an FeSOD. The recombinant enzyme also protected the E. coli mutant QC779 from paraquat toxicity. Northern blot analysis showed that FeSODB is differentially expressed, showing a higher level at the epimastigote stage of T. cruzi development; whereas, FeSODA is constitutively expressed at a lower level in all developmental stages. Furthermore, Southern hybridization shows that both FeSODA and FeSODB genes appear to be present in the T. cruzi genome as multiple repeating units (multi-copy gene family). PMID- 9200126 TI - Three main patterns in the expression of six actin genes in the plerocercoid and adult Diphyllobothrium dendriticum tapeworm (Cestoda). AB - The expression of six actin genes was examined in adult and plerocercoid Diphyllobothrium dendriticum tapeworms using in situ hybridization. On the basis of their structures, these genes are divided into three groups, the cestoda-I, II and -III actins. Current studies show that the expression of actins belonging to different groups vary to a great extent. The three cestoda-I actins are expressed primarily in muscle cells of both adult and plerocercoid tapeworms, the expression being restricted to fewer cells in the plerocercoid larva. The two cestoda-II actins are cytoplasmic actin isoforms, expressed in a variety of cells, i.e. in cells dividing, differentiating and migrating. Expression of the cestoda-III actin gene is detected merely in the peripheral part of the outer parenchyma, mainly in the tegument cell bodies. This pattern is very weak in plerocercoids. The results indicate that actins also in D. dendriticum can be divided into cytoplasmic and muscle-specific isoforms. In this organism, one major pattern of muscle actin gene expression (cestoda-I) and two major patterns of non-muscle actin gene expression (cestoda-II and -III) were found. PMID- 9200127 TI - Trypanosoma cruzi strains partition into two groups based on the structure and function of the spliced leader RNA and rRNA gene promoters. AB - We have previously identified a major proximal sequence element (PSE) responsible for transcription of the spliced leader (SL) gene from Trypanosoma cruzi strain CL, and showed that the sequence encompassing this PSE exhibits approximately 30% divergence between two major groups of T. cruzi isolates, but strong conservation within the groups. In this report, we show that the SL RNA gene promoter from the CL strain (group I) is efficiently expressed only in T. cruzi isolates from group I. Similarly, the sequence of the approximately 643 bp promoter region of the T. cruzi rRNA is strongly conserved within, but diverged approximately 20% between, the two groups. Reporter constructs driven by the rRNA promoter sequences from group I strains are strongly expressed after electroporation into other group I strains, but not expressed in group II strains. In contrast, constructs bearing rRNA promoter sequences from group II strains are active in strains from both groups. Phylogenetic analyses performed with both the rRNA and the SL RNA gene promoter sequences yielded similar trees, and these trees strongly reinforce the partitioning of known T. cruzi into two major groups that parallel the observed functional specificity of the promoters. Given the well-documented species specific pattern of both rRNA promoters and PSEs in higher eukaryotes, these results suggest an ancient evolutionary divergence among organisms currently classified as T. cruzi. PMID- 9200128 TI - Diversity in repeat-containing surface proteins of Leishmania major. AB - The gene B protein (GBP) is one of the products of the LmcDNA16 gene family, a cluster of related but non-identical genes that are differentially-expressed during the Leishmania life cycle. This protein, which is found on the surface of infective stage parasites, contains an extensive region of proline-rich amino acid repeats, constituting 45% of the total protein. The structure and stability of these repeats have been investigated in a number of L. major strains by polymerase chain reaction (PCR) amplification and Southern blotting. Data reported in this paper demonstrate variability between strains with respect to the number of repeats encoded by GBP, although those strains isolated within adjacent geographical regions have conserved repeat structures. The data also reveal that some parasite lines have additional repeat sequences within a second, related gene in the LmcDNA16 array. Western blotting experiments have established that these sequences are expressed in vivo, indicating that L. major strains are heterogeneous in their surface complement of gene B repeat-containing proteins. PMID- 9200129 TI - Structural characterization of the N-glycans from Echinococcus granulosus hydatid cyst membrane and protoscoleces. AB - Infection by the tapeworm Echinococcus granulosus in the intermediate host results in the development of a hydatid cyst which contains the protoscoleces within a fluid-filled cavity enclosed by the bilayered cyst membrane. N-glycans were enzymatically released from crude extracts of homogenates of hydatid cyst membranes and protoscoleces and their structures were defined by high sensitivity fast atom bombardment mass spectrometry in conjunction with sequential exoglycosidase digestions. The major N-glycans from the cyst membrane were found to be non-charged structures having complex-type antennae and core fucosylation. The antennae are either truncated at the first N-acetylglucosamine or are extended with beta-galactose to form N-acetyllactosamine (lacNAc). A significant proportion of the lacNAc backbones are capped by alpha-galactose. The resulting Gal alpha-Gal beta-terminal structures may account for the earlier observation that antibodies against the blood group P1 epitope recognise components of hydatid cyst extracts. The complex-type N-glycans identified in the protoscoleces extracts were the same as the neutral structures found in the cyst membrane but a small proportion of high mannose structures and truncated di- and trimannosyl core structures were also identified. Sialylated N-glycans were identified as minor constituents of the cyst membrane preparation but were not observed in protoscoleces extracts. Whether the sialylated glycans are host derived or endogenously synthesized by the parasite remains to be established. This is the first reported structural analysis of N-glycans from cestodes and provides new insights into protein glycosylation in helminths. PMID- 9200131 TI - Amino acid distribution in immature rat brain. AB - We compared the levels of amino acids in the free pool in 6 regions (cerebral cortex, olfactory bulb, substantia nigra, globus pallidus, caudate nucleus, and spinal cord) in the newborn rat brain. The amino acid distribution was heterogeneous, with the area of highest concentration containing 2-3 fold as much as the lowest area. These differences were considerably less than those previously found for adult brain. Although some areas often contained high levels of amino acids, and others mostly low levels, the distribution of the various amino acids was highly variable. This heterogeneity of distribution in the newborn brain was different from that in the adult brain. We conclude that there is significant heterogeneity of amino acid distribution, that it is different for each amino acid, and that it undergoes major changes during development. PMID- 9200130 TI - Sequence diversity in the amino-terminal 47 kDa fragment of the Plasmodium falciparum serine repeat antigen. PMID- 9200132 TI - Some doubts about the basic concept of hole-board test. AB - Hole board test is a generally used method for screening the potential anxiolytic character of drugs. The test is based on the assumption, that head-dipping activity of the animals is inversely proportional to their anxiety state. We tested this assumption by measuring the head-dipping activity of animals in environments with different levels of aversive character (illumination). The anxiolytic chlordiazepoxide significantly elevated head-dipping activity in moderately aversive environment, it was not active in non-aversive environment, and it exhibited inhibitory activity in highly aversive environment. When the latency of the first head-dip was measured, we found that the proportion of animals with short latency was significantly increased in moderately and highly aversive environments. It is concluded, that the inverse relation between anxiety state and head-dipping activity is true only in a certain range of anxiety level. In more aversive situations, when the anxiety level of the animals is high, the holes nay represent a possible way to escape from the aversive environment instead of an explorable object. In this case the relation between anxiety state and head-dipping activity is directly and not inversely proportional. PMID- 9200133 TI - Tau proteins bind to kinesin and modulate its activation by microtubules. AB - Microtubule-associated tau proteins are likely candidates to interfere with axonal transport of membranous organelles. We studied that tau proteins influenced the enzyme activity of kinesin, known to drive anterograd transport along microtubules. An in vitro reconstituted system was applied; microtubules were assembled from purified tubulin with or without tau proteins. Both types of reconstituted microtubules stimulated MgATPase activity of purified kinesin in a concentration dependent, saturable manner. The extent of maximal stimulation by tau-coated microtubules was lower than that of microtubules without tau proteins. Analysis of kinetic data, on the other hand, suggests that tau-coated microtubules apparently bind kinesin with higher affinity then microtubules not associated with tau proteins. Tau proteins, similarly to tubulin dimers, seem to bind to the heavy chain of kinesin. These data support the notion that tau proteins could act as regulators of kinesin-driven processes. PMID- 9200134 TI - Opposite short-term changes induced by an organophosphate in cortical and hippocampal evoked activity. AB - Organophosphates are the most widely used pesticides throughout the world. The considerable amount brought out to the environment poses a risk on the whole population. As organophosphates are neurotoxic substances and their residues can persist in the environment for several weeks, their influence on the nervous system of humans and animals is of principal interest. In the present study, we investigated the alterations induced by dichlorvos, a common pesticide substance, in parameters of somatosensory evoked potentials and hippocampal evoked population spikes of rats. The changes of the cortical vs. hippocampal evoked responses were opposite and only hippocampal effects could be directly explained through an increased cholinergic activity. PMID- 9200135 TI - Quantitative analysis of gyrification of cerebral cortex in dogs. AB - The degree of gyrification of the cerebral cortex was studied in a population of various breeds of dogs. The aim of the study was to demonstrate whether or not variations in brain weight, body size or body weight affect gyrification within a species. Results suggest that in spite of significant correlations between all these parameters the degree of gyrification is essentially determined by the size of the brain. This parameter shows neither sex-dependence nor asymmetry between the two hemispheres. The examination of frontal serial sections through dogs' brains has also revealed that in agreement with data on the primate brain the association cortical regions show an extraordinarily high gyrification. Accordingly, gyification is dependent from the overall growth of the brain in particular from that of the association areas growing most intensively in mammals. Measurement of the gyrification index is therefore, thought to be a suitable method to express in quantitative terms developmental trends of cortical organization. PMID- 9200136 TI - Retinoic acid induces a tissue-specific deletion in the expression domain of Otx2. AB - The expression domain of Otx2, a gene essential for the development of the fore- and midbrain, has previously been shown to be affected by exposure to all-trans retinoic acid (AT-RA). However, morphological abnormalities of the fore- and midbrain induced by exposure of early somite-stage embryos to AT-RA were not associated with abnormal Otx2 expression. To identify abnormal expression of developmental genes induced by exposure at early somite-stages, we performed a fine analysis of the expression domains of Otx2, Otx1, Emx2, and Pax-6 by combining in situ hybridization (ISH) with computer-assisted superpositions and three-dimensional reconstructions of these expression domains. No alteration in the relative location of the caudal boundaries of the expression domains of these genes was observed. The only abnormality was a deletion of the most cranial portion of the neural folds (NF). PMID- 9200137 TI - Reading and language in 9- to 12-year olds prenatally exposed to cigarettes and marijuana. AB - Facets of reading and language were examined in 131 9- to 12-year-old children for whom prenatal exposure to marijuana and cigarettes had been ascertained. The subjects were from a low-risk, predominantly middle class sample who are participants in an ongoing longitudinal study. Discriminant Function Analysis revealed a dose-dependent association that remained after controlling for potential confounds, between prenatal cigarette exposure and lower language and lower reading scores, particularly on auditory-related aspects of this latter measure. The findings are interpreted as consistent with earlier observations of an association between cigarette smoking during pregnancy and altered auditory functioning in the offspring. Similarities and differences between the reading observations and dyslexia are discussed. Maternal prenatal passive smoke exposure did not appear to contribute to either the language or reading outcomes at this age but postnatal secondhand smoke exposure by the child was associated with poorer language scores. Prenatal marijuana exposure was not significantly related to either the reading or language outcomes. PMID- 9200138 TI - Effects of prenatal ethanol exposure and early experience on radial maze performance and conditioned taste aversion in mice. AB - C57BL/6 mice were intubated on gestational days 14-18 twice daily with 1.58 g/kg ethanol, 4.2 g/kg sucrose, or remained untreated. Offspring of ethanol-treated or lab chow control groups were raised either by group-housed dams and weaned on postnatal day (PND) 28 (enriched condition), or by individually housed dams and weaned on PND 21 (standard condition). Offspring of the sucrose control group were raised by individually housed dams and weaned on PND 21. Groups did not differ in pup weight or litter size. Male and female offspring were assessed for performance in an unbaited radial maze (PND 45-52) and male offspring only were tested for conditioned taste aversion (PND 54-59). As hypothesized, mice prenatally exposed to ethanol and raised under standard conditions failed to develop the conditioned taste aversion response. In contrast, subjects with in utero ethanol exposure that were raised under enriched preweaning conditions developed the taste aversion response. Maze performance improved significantly over days, but no significant effects were detected for either prenatal treatment or preweaning rearing conditions. In conclusion, enriched preweaning rearing conditions abolished the detrimental effects of prenatal ethanol exposure on conditioned taste aversion, but radial maze performance remained unaffected by any treatment in this study. PMID- 9200139 TI - Paternal exposure of rabbits to lead: behavioral deficits in offspring. AB - Paternal exposures to exogenous agents have been reported to produce a variety of developmental defects in the offspring. In experimental animals, these effects include decreased litter size and weight, increased stillbirth and neonatal death, birth defects, tumors, and functional/behavioral abnormalities-some of these effects being transmitted to the second and third generations. The majority of experimental studies assessing nervous system function of offspring following paternal exposures have utilized rats as the experimental animal, but other species can be used. The National Toxicology Program (NTP) has initiated studies to validate the rabbit as an animal model for human reproductive toxicity, because rabbits are the smallest laboratory animal from which ejaculates can be collected repeatedly. An important part of reproductive toxicology is assessment of the reproductive ability of males following exposure, as well as developmental and functional assessment of their offspring. This article describes a pilot study and a main study to investigate the feasibility of using rabbits to assess the functional effects of paternal exposure to lead. The pilot study included seven male rabbits per group exposed for 15 weeks to lead acetate sufficient to produce 0, 50, or 110 micrograms/dl blood lead. The main study included 15 male rabbits per group exposed for 15 weeks to lead acetate to produce 0, 20, 40, and 80 micrograms/dl blood lead. At the conclusion of the exposure, male rabbits were mated with unexposed females. These females carried their litters to term, delivered, and reared their own offspring. The offspring were weighed at 5, 10, 15, 20, 25, 30, and some at 35 days of age. They were also tested for exploratory activity in a standard figure-eight "maze" for 30 min/day on days 15, 20, 25, and 30. A second assessment of exploratory behavior, along with a simple test of aversive conditioning, was attempted in the pilot study, but was judged not to be suitable for the main study. Of the 21 male rabbits that were mated in the pilot study, 16 produced viable litters (6/7, 6/7, and 4/7 in control, low- and high lead groups, respectively), with a mean number of 6 live births/litter in each treatment group (range 2-8). Of the 60 rabbits mated in the main study, 57 produced litters, and two rabbits died giving birth. Significant postnatal deaths were observed in all groups, with about one half of the offspring dying before testing was initiated at day 15. There were no treatment-related effects on offspring weight gain through wearing. The data suggest that paternal lead exposure of rabbits may reduce figure-eight activity on day 25, the time of peak activity in the offspring. PMID- 9200140 TI - Retinal lesions in rat fetuses prenatally exposed to cocaine. AB - The increased use of cocaine in the United States and worldwide has created great concern about its effects on fetuses and neonates of pregnant cocaine abusers. The effects on neonates are varied: fetal growth delay, microcephaly, abnormal neurological functions, microphthalmia, and maternal obstetric complications. In this study, the effect of prenatal cocaine administration was studied microscopically in the retina of rat fetuses. Twenty-five pregnant Wistar rats were injected i.p. with an aqueous cocaine solution using a 30 mg/kg daily dose for 45 days. Control group rats (15 pregnant animals) received saline solution for the same period. Day 0 of gestation was the day after mating. Dosing began on this day. The rats were killed on gestation day 21 and fetuses were obtained for examination. The histopathological light and electron microscope studies of the retinas showed interstitial oedema, areas depleted of cells, necrosis, and hyperchromatic ganglion cells. There also was a significantly lower number of retinal cells compared to control fetuses. In four cases, teratogenic lesions of the retina were observed whereas no changes were present in control fetuses. Results indicate that development of retina in fetuses prenatally exposed to cocaine was altered by cocaine exposure. PMID- 9200141 TI - Effects of low-dose phenytoin administered to newborn mice on developing cerebellum. AB - To examine correlations between dose levels of phenytoin (PHT) and neurotoxic effects on cerebellar development, we administered 10, 17.5, 25, and 35 mg/kg PHT suspended in sesame oil orally to newborn Jcl:ICR mice once a day during postnatal days 2-4 and determined plasma PHT concentrations during the administration period. Mortality rates were 12.5% and 35.2% in males and 15.3% and 33.3% in females for the 25 and 35 mg/kg PHT-treated groups during the PHT treatment, respectively. In the 25 and 35 mg/kg PHT-treated groups, total brain weight, the size of the cerebellum, and cerebellar weight were significantly reduced on postnatal day 21. However, in the 10 and 17.5 mg/kg PHT-treated groups, total brain weight and the size and weight of the cerebellum did not differ from those of the control group. Histologically, the number of pyknotic cells in the external granular layer (EGL) in the 25 and 35 mg/kg PHT-treated groups was increased on postnatal day 5, and the EGL was thicker than in the control group on postnatal day 14. Some of the Purkinje cells in the 35 mg/kg PHT treated group showed degeneration. Plasma PHT levels were 10.7 +/- 2.2 and 24.6 +/- 2.6 micrograms/ml in the 25 and 35 mg/kg PHT groups on the third day of PHT treatment, respectively. In the 25 mg/kg PHT group, plasma PHT level was found to be in the therapeutic range for humans, 10-20 micrograms/ml. Accordingly, during pregnancy, epileptic women should be carefully given PHT at the lowest effective dose while plasma PHT levels are monitored properly. These findings emphasize the importance of pharmacokinetics in evaluating of phenytoin-induced developmental neurotoxicity. PMID- 9200142 TI - The effect of developmental exposure to cadmium (Cd) on visual evoked potentials (VEPs) and lipid peroxidation. AB - Pregnant Swiss albino rats were divided into three groups: control (C), gestational exposure of Cd (G-Cd), and gestational/postnatal exposure of Cd (GP Cd) groups. Control animals received tap water, and the rats of GP-Cd group received Cd as CdC12 in their drinking water during the experimental period. The G-Cd group was given Cd during pregnancy, but given tap water after birth. Twenty two days after birth, 15 rats (for each group) were taken from their mothers and continued to be treated with Cd (GP-Cd group) or tap water (C and G-Cd groups) for an additional 38 days. On postnatal day (PND) 60, flash visual evoked potentials (FVEPs) were recorded with disc electrodes attached with collodion 0.5 cm in front of and behind bregma. The mean latencies on N1, P2, and P3 were prolonged in the GP-Cd group compared with controls. The mean latency of P3 was also significantly different between G-Cd and GP-Cd groups. P1-N1 and N1-P2 amplitudes of VEPs were significantly decreased in the GP-Cd group compared with control group. N1-P2 amplitude of the G-Cd group was significantly lower than that of the control group. Thiobarbituric acid reactive substances (TBARS) were determined as an indicator of lipid peroxidation. Our data showed that pre- and postnatal Cd treatment caused a significant increase of lipid peroxidation in the brain. PMID- 9200143 TI - S-allyl cysteine inhibits nitrosomorpholine formation and bioactivation. AB - Water extracts of garlic, deodorized garlic powder, and onions, but not leeks, were found to significantly (p < 0.05) reduce the in vitro formation of N nitrosomorpholine (NMOR), a known liver carcinogen. Addition of increasing quantities (20, 40, and 80 mM) of S-allyl cysteine (SAC), a water-soluble compound in processed garlic, depressed NMOR formation by 16%, 27%, and 43%, respectively (p < 0.05). The ability of SAC to block NMOR formation decreased as the NaNO7 and morpholine concentrations increased. SAC and its non-allyl analog S propyl cysteine effectively blocked NMOR formation. SAC and S-propyl cysteine were less effective than isomolar cysteine in reducing NMOR formation (p < 0.05). The oil-soluble sulfur compounds diallyl disulfide (DADS), dipropyl disulfide, and diallyl sulfide were ineffective inhibitors of NMOR generation (p > 0.05). SAC and DADS reduced the mutagenicity of NMOR in Salmonella typhimurium TA100 (p < 0.05). SAC at 70 mumol/plate reduced the number of histidine revertants per plate by 51% (p < 0.05), whereas DADS at 0.12 mumol/plate reduced mutant colony number by 76% (p < 0.05). SAC and DADS were more effective than isomolar cysteine in reducing NMOR mutagenicity (p < 0.05). The ability of sulfur compounds in garlic and onions to depress nitrosamine formation and bioactivation in these studies is consistent with epidemiologic evidence that higher intake of allium plants is associated with a reduction in the risks of some cancers. PMID- 9200144 TI - Comparison of pure inositol hexaphosphate and high-bran diet in the prevention of DMBA-induced rat mammary carcinogenesis. AB - Inositol hexaphosphate (IP6), abundant in cereals and legumes, has been demonstrated to be a promising anticancer agent in different in vivo and in vitro models. Because IP6 is particularly abundant in the bran part of certain mature seeds such as wheat, we investigated whether a high-fiber bran diet containing high IP6 shows a dose-response inhibition of 7,12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary carcinogenesis. Starting at two weeks before DMBA intubation, rats were divided into five groups and fed AIN-76A diet only or AIN 76A diet containing 5%, 10%, or 20% Kelloggs' All Bran; the fifth group received 0.4% IP6 given in drinking water, an amount equivalent to the IP6 content in 20% bran. After carcinogen administration, the rats remained on these regimens for 29 weeks. Compared with the carcinogen control, at 29th week, tumor incidence was reduced by 16.7%, 14.6%, and 11.4% in rats fed 5%, 10%, and 20% bran, respectively (not statistically significant). However, rats given 0.4% IP6 in drinking water, equivalent to that in 20% bran, had a 33.5% reduction in tumor incidence (p < 0.02) and 48.8% fewer tumors (p < 0.03). These data show that supplemental dietary fiber in the form of bran exhibited a very modest, statistically nonsignificant inhibitory effect, which was also not dose dependent. In contrast, animals given IP6 showed significant reduction in tumor number, incidence, and multiplicity. Thus IP6 an active substance responsible for cereal's beneficial anticancer effect, is clearly more effective than 20% bran in the diet. In practical terms, intake of IP6 may be a more pragmatic approach than gorging enormous quantities of fiber for cancer prophylaxis. PMID- 9200145 TI - Dietary fiber and risk of breast cancer: a case-control study in Uruguay. AB - To examine whether dietary fiber modifies breast cancer risk, a case-control study involving 351 newly diagnosed patients with breast cancer and 356 hospitalized controls was conducted in Uruguay. Dietary patterns were assessed in detail by use of a food frequency questionnaire on 64 items, which allowed the calculation of total energy intake. Nutrient residuals were calculated through regression analysis. After adjustment for potential confounders (which included age, residence, family history of breast cancer, prior history of benign breast disease, parity, total energy, red meat, lutein/zeaxanthin and quercetin intake, and menopausal status), dietary fiber and total nonstarch polysaccharides were associated with a strong reduction in risk of breast cancer (odds ratio for uppermost quartile of total dietary fiber = 0.51, 95% confidence limit = 0.31 0.82). Also the dose-response pattern was highly significant (p < 0.001). The inverse association was observed in pre- and post-menopausal women and was similar for soluble and insoluble fiber. Furthermore, dietary fiber displayed a strong joint effect with fat, quercetin, and lutein/zeaxanthin. PMID- 9200147 TI - dl-alpha-tocopherol induces apoptosis in erythroleukemia, prostate, and breast cancer cells. AB - Vitamin E, best known as a potent antioxidant, has been shown to have other functions that are not mediated by this activity. Recent reports have suggested that vitamin E may inhibit smooth muscle cell and also cancer cell growth. We have studied the effect of dl-alpha-tocopherol (vitamin E) on a series of well established cancer cell lines that included two erythroleukemia cell lines and a hormone-responsive breast and prostate cancer cell line. Cell proliferation was examined in these cell lines, which were maintained at optimal growth conditions. A dose-dependent inhibition of cell growth was found in all cell lines examined, with the MCF-7 breast and CRL-1740 prostate cancer cell lines showing potent suppression of growth at 0.1 mM vitamin E, whereas the erythroleukemia cell lines, HEL and OCIM-1, responded only at > 0.25 mM vitamin E with inhibition of proliferation. Studies of [3H]thymidine incorporation showed that vitamin E supplementation reduced DNA synthesis in all cell lines. Analysis of high molecular-weight DNA revealed extensive fragmentation, indicating apoptosis of all cell lines supplemented with vitamin E. Our studies thus give evidence of a general inhibition of cell proliferation by dl-alpha-tocopherol, with breast and prostate cancer cells distinctly more sensitive than erythroleukemia cells. PMID- 9200146 TI - Relationship among colonocyte proliferation, differentiation, and apoptosis as a function of diet and carcinogen. AB - To determine the relationship among colonocyte proliferation, differentiation, and apoptosis as a function of fiber and carcinogen, we conducted a 2 x 2 factorial design study with two fibers (pectin or cellulose) and two injection protocols (azoxymethane or saline) in male Sprague-Dawley rats. Rats were killed six weeks after the injections, and in vivo cell proliferation was measured by incorporation of bromodeoxyuridine into DNA, differentiation by binding of the lectin Dolichos biflorus agglutinin, and apoptosis by immunoperoxidase detection of digoxigenin-labeled genomic DNA. In the proximal colon, pectin decreased differentiation and apoptosis, resulting in a greater number of cells per crypt column. In the distal colon, pectin increased cell proliferation, resulting in more crypts per millimeter of colon and a greater number of surface cells. Azoxymethane increased cell proliferation and decreased differentiation and apoptosis in the proximal and the distal colon. This resulted in a greater number of surface cells proximally and more crypts per millimeter of colon distally. These results illustrate the importance of considering all three parameters (proliferation, differentiation, and apoptosis) when evaluating neoplastic growth. PMID- 9200148 TI - Increasing dietary lipid and iron content decreases manganese superoxide dismutase activity in colonic mucosa. AB - Manganese superoxide dismutase (MnSOD) is an important mitochondrial antioxidant. Alteration in the regulation of MnSOD activity has been proposed to play a critical role in the development of many types of tumors. Colorectal cancer is one of the most common human malignancies and has been shown to be influenced by dietary factors. Two such dietary factors include lipid and iron. Lipid and iron are also potential modulators of MnSOD activity. This study examined lipid and iron influence on MnSOD activity in colonic mucosa. Fischer rats were fed one of eight test diets for six weeks. Four of the diets included AIN-76A-based formulas containing 5% corn oil or 20% lipid from corn oil, menhaden oil, or beef tallow. Four additional diets included identical formulations with iron supplemented to a level of 140 mg/kg. Results showed that iron supplementation decreased MnSOD activity in animals fed the 5% corn oil diet. An increase in dietary lipids from 5% to 20% also decreased MnSOD activity in colonic mucosa. The lipid and iron variables used in this study decreased MnSOD activity without affecting manganese status or other antioxidant mechanisms in this tissue. PMID- 9200149 TI - Influence of isoflavones in soy protein isolates on development of induced prostate-related cancers in L-W rats. AB - Lobund-Wistar (L-W) rats are inherently susceptible to spontaneous and induced metastasizing adenocarcinomas in the prostate-seminal vesicle (P-SV) complex. L-W rats were fed soy protein isolates containing high isoflavones (genistein and daidzein) or low isoflavones to determine their effects on development of induced P-SV tumors in two stages of the tumorigenic process. In rats fed the high isoflavone-supplemented soy diet before initiation by methylnitrosourea (MNU), the incidence of induced prostate-related cancer was reduced and the disease-free period was prolonged by 27% compared with rats fed the same diet but low in isoflavones. Rats fed the same diets, started after MNU, manifested suggestive but less consistent results than those noted above. The incidence rates were of marginal significance, suggesting that the high intensity of the active induced disease may not represent the character of the slower-growing spontaneous (natural) disease. The delay of disease onset is of clinical significance. PMID- 9200150 TI - Nutrient intake according to education, smoking, and alcohol in Italian women. AB - The control group of a hospital-based case-control study on breast cancer was used to assess the relationships between education, smoking habits, alcohol consumption, and intake of selected macro- and micronutrients in Italian women. The study subjects were 2,588 women admitted to a network of hospitals in various Italian regions for nonneoplastic, acute diseases unrelated to long-term changes in the diet. Although relatively few differences were observed, less educated subjects consumed more linoleic acid and polyunsaturated fats than did more educated women. Smoking habits were associated with the largest differences in selected antioxidant vitamins. Significant differences were observed for beta carotene and vitamin C intake, with an 11% higher intake of beta-carotene and a 12% higher intake of vitamin C in ex-smokers than in current smokers. Heavier alcohol drinkers tended to consume more retinol and iron but less beta-carotene than did moderate or nondrinkers. Thus the differences in macro- and micronutrient intake were generally moderate across categories of education, smoking, and alcohol consumption in this data set of Italian women. Nonetheless, they confirm the importance of allowing for these variables in analyzing the relationship between nutritional factors and disease risk. PMID- 9200151 TI - Are dietary factors involved in DNA methylation associated with colon cancer? AB - Disturbances in DNA methylation have been hypothesized as being involved in carcinogenesis. It has been proposed that dietary factors such as folate, alcohol, and methionine may be associated with colon cancer because of their involvement in DNA methylation processes. Data from a large retrospective population-based case-control study of incident colon cancer were used to evaluate whether intake of alcohol and other dietary factors involved in DNA methylation are associated with colon cancer. Dietary data were obtained using a detailed diet history questionnaire. We did not observe strong independent associations between folate, vitamin B6, vitamin B12, methionine, or alcohol and risk of colon cancer after adjusting for body size, physical activity, cigarette smoking patterns, energy intake, and dietary intake of fiber and calcium. However, when assessing the associations between colon cancer and a composite dietary profile based on alcohol intake, methionine, folate, vitamin B12, and vitamin B6, we observed a trend of increasing risk as one moved from a low- to a high-risk group. This trend was modest and most marked in those diagnosed at a younger age [odds ratio (OR) for men = 1.3, 95% confidence interval (CI) = 0.9 1.9; OR for women = 1.6, 95% CI = 1.0-2.6]. We observed that associations with this high-risk dietary profile were greater among those who took aspirin or nonsteroidal anti-inflammatory drugs on a regular basis and were younger at the time of diagnosis (men OR = 1.7, 95% CI = 1.0-3.2; women OR = 2.2, 95% CI = 1.0 4.8) and for distal tumors (men OR = 1.4, 95% CI = 0.9-2.3; women OR = 2.0, 95% CI = 1.0-3.8). Findings from this study provide only limited support for previously reported associations between dietary factors involved in DNA methylation and risk of colon cancer. PMID- 9200152 TI - Intervention of transplantable human mammary carcinoma MX-1 chemotherapy with dietary menhaden oil in athymic mice: increased therapeutic effects and decreased toxicity of cyclophosphamide. AB - We investigated the effects of dietary menhaden oil on cyclophosphamide (CP) antineoplastic activity and its protective effect against CP toxicity. We found that dietary menhaden oil (HMO, 20% menhaden oil + 5% corn oil) enhanced the CP antitumor effect at the lowest dose tested (50 mg/kg) compared with the control group (LCO, 5% corn oil). Dietary HMO and CP treatment had a significant effect on the activities of tumor and liver microsomal cytochrome P-450 (CYP) over the controls. Activity of one of the key CP activating enzymes, CYP2B1 (which is similar to human CYP2B6), was significantly enhanced in the liver and tumor by the HMO diet, which could result in the formation of more pharmacologically active CP metabolites and, therefore, increased CP antitumor response. Moreover, the HMO diet exhibited a very significant protective effect against CP acute toxicity. The activity of the CP detoxifying enzyme aldehyde dehydrogenase (ADH) was significantly increased in the liver after HMO feeding; thus the observed protective effect of HMO feeding against CP toxicity may be partially the result of induction of ADH activity in the liver. In summary, our findings suggested that dietary menhaden oil can modulate ADH and CYP activities in a manner that may alter the metabolism of CP and, therefore, improve its therapeutic index by increasing its therapeutic effect and decreasing its toxicity. PMID- 9200153 TI - Possible mechanisms involved in apoptosis of colon tumor cell lines induced by deoxycholic acid, short-chain fatty acids, and their mixtures. AB - Apoptosis of tumor cells is an important growth-regulating event in tumor masses. In this study we have confirmed that deoxycholic acid (DCA) and the short-chain fatty acids (SCFA) butyrate and propionate induce a time- and concentration dependent apoptosis in two human colon tumor cell lines: HT-29 and CaCO2. DCA is more potent, inducing effects at low concentration (50 microM) and after 24 hours of incubation, whereas SCFA (4 mM) requires 72-96 hours of treatment. Combining low concentrations of DCA (12.5-25 microM) with butyrate and propionate (4 mM) produces an additive effect on the percentage of apoptotic cells, as demonstrated by flow cytometry and DNA fragmentation. Protein kinase C, protein tyrosine kinase, and gene transcription/translation inhibitors do not significantly modify the rate of apoptosis, whereas the intracellular Ca2+ chelator 1,2-bis(o aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA-AM) completely abolishes the DCA-induced effect without affecting the SCFA-induced apoptosis. Measurement of intracellular Ca2+ by inverted fluorescence microscopy reveals that DCA induces a rapid increase of cytosolic Ca2+ that is abolished when the cells are preincubated with BAPTA-AM, whereas ethyleneglycolbis(beta aminoethyl ether)-N,N,N',N'-tetraacetic acid has a minimal effect. In contrast, SCFA does not modify the intracellular Ca2+ concentration. Thus the DCA-induced apoptosis is a Ca(2+)-dependent process, whereas the intracellular signals responsible for the SCFA-induced effect remain unknown. The ionophore activity of DCA could be responsible for the increased intracellular Ca2+, but other mechanisms, such as activation of phospholipase C and phosphoinositide hydrolysis, have to be considered. PMID- 9200155 TI - Trends in food intake: the 1987 and 1992 National Health Interview Surveys. AB - To examine food intake trends in the US population, cross-sectional nationally representative food intake data were obtained from the 1987 and 1992 National Health Interview Survey Cancer Control Supplements. In each of these years, approximately 10,000 respondents completed methodologically consistent food frequency questionnaires containing the same 57 food items. Between 1987 and 1992, the proportion of Americans consuming high-fat foods, including fried fish, fried chicken, bacon, eggs, whole milk, and butter, decreased. The proportion of Americans drinking alcoholic beverages also decreased: fewer drank wine and hard liquor in 1992. The proportion of fruit and vegetable consumers remained stable over time. These results are similar to those obtained from more detailed national surveys. National guidelines urge Americans to avoid intake of high-fat foods, increase consumption of fruits and vegetables, and practice moderation when drinking alcoholic beverages to prevent cancer and other chronic diseases. The direction of Americans' apparent changes in food usage between 1987 and 1992, evaluated using limited data from food frequency questionnaires, suggests greater behavioral changes in the direction of guidelines recommending avoidance of foods that may increase the risk of cancer than in the direction of guidelines recommending increased consumption of foods that may confer protection. PMID- 9200154 TI - Suppression of aberrant colonic crypt foci by synthetic sphingomyelins with saturated or unsaturated sphingoid base backbones. AB - Supplementation of the diet of CF1 mice with sphingomyelin isolated from milk has been shown to reduce the number of aberrant crypt foci (ACF) and the appearance of colonic adenocarcinoma induced by 1,2-dimethylhydrazine (Schmelz et al., Cancer Res 56, 4936-4941, 1996). The objective of this study was to determine whether chemically synthesized sphingomyelin reduces the appearance of ACF, one of the earliest morphological changes in the development of colonic tumors, and to investigate the specificity of this inhibition for the unsaturated sphingoid base backbone. 1,2-Dimethylhydrazine was administered intraperitoneally to female CF1 mice, then the animals were fed a semipurified AIN 76A diet without supplementation (controls) or supplemented with 0.1% (wt/wt) sphingomyelin isolated from skim milk powder, synthetic N-palmitoylsphingomyelin, or N palmitoyldihydrosphingomyelin for four weeks. The number of ACF in the sphingomyelin-fed groups was significantly lower than in the control by 54% (p = 0.002), 52% (p = 0.002), and 70% (p < 0.0001) for milk sphingomyelin, synthetic sphingomyelin, and synthetic dihydrosphingomyelin, respectively. Suppression of ACF by the synthetic dihydrosphingomyelin was significantly greater than by synthetic sphingomyelin (p = 0.035). These findings establish that sphingomyelin, and not merely a possible contaminant of the naturally occurring sphingomyelin preparation used previously, suppresses ACF formation. Furthermore, the greater potency of dihydrosphingomyelin reveals that the 4,5-trans double bond of the sphingoid backbone is not required for this suppression. PMID- 9200156 TI - Antiproliferative effect of fermented milk on the growth of a human breast cancer cell line. AB - In vivo and in vitro studies have shown an antitumor activity of Lactobacilli in colon cancer, and some epidemiologic studies have indicated a reduced risk of breast cancer in women who consume fermented milk products. We studied the direct effect of milk fermented by five bacteria strains (Bifidobacterium infantis, Bifidobacterium bifidum, Bifidobacterium animalis, Lactobacillus acidophilus, and Lactobacillus paracasei) on the growth of the MCF7 breast cancer cell line. Our results showed a growth inhibition induced by all fermented milks, even though B. infantis and L. acidophilus were the most effective (85% inhibition after 9 days). The antiproliferative effect was not related to the presence of bacteria in fermented milk, and neither whole milk (crude or ultrahigh temperature sterlizied) nor its main fractions (lactalbumin or beta-lactoglobulin fraction) affected cell growth. Our findings suggest the presence of an ex novo soluble compound produced by lactic acid bacteria during milk fermentation or the microbial transformation of some milk components in a biologically active form. Although the mechanism of the antitumor activity is not clear, the present study suggests the potentiality offered by fermented milk as producers of compounds with antiproliferative activity useful in the prevention and therapy of solid tumors like breast cancer. PMID- 9200157 TI - Effect of acute acipimox administration on the rates of lipid and glycogen synthesis in cachectic tumor-bearing rats. AB - Increased energy expenditure in cancer cachexia may be associated with increased postprandial glycogen synthesis via an indirect pathway involving gluconeogenesis. The possible beneficial effect of acipimox, a nicotinic acid analogue that suppresses lipolysis and may also inhibit gluconeogenesis, were therefore examined. Rats bearing a transplantable Leydig cell tumor and freely fed controls were fasted overnight, then given a test meal with or without 10 mg of acipimox. The meal included 200 mg of [1-13C]glucose, and the rats were injected simultaneously with 7 mCi of 3H2O and 1 microCi of [14C]glycerol. The rats were killed one hour later. The rate of incorporation of 3H2O into hepatic glycogen was increased in the tumor-bearing rats and suppressed by acipimox. Positional analysis of the tritium incorporated into glycogen indicated that a greater proportion of the glycogen was synthesized via pyruvate in the tumor bearing rats. Acipimox tended to reduce this proportion, although the effect was not statistically significant. Neither tumor growth nor acipimox significantly affected the proportion of 13C incorporated into different positions in the glycogen glucose. Glycogen synthesis from glycerol tended to decrease when lipolysis was suppressed by acipimox, although the statistical significance of this effect was marginal. Fatty acid synthesis in liver and adipose tissue was reduced in tumor-bearing rats, but acipimox had no effect. It is concluded that acipimox does suppress gluconeogenesis and glycogenesis in the postprandial state, but it does not normalize all the metabolic abnormalities observed in cancer cachexia. PMID- 9200158 TI - Effects of caloric intake on anticancer therapy in rats with valine-depleted amino acid imbalance. AB - Valine-depleted amino acid imbalance solution markedly inhibits tumor growth but causes fatty liver as a side effect. However, much remains unknown about the mechanism of the development of fatty liver. Valine-depleted amino acid imbalance solution containing various concentrations of calories was administered to tumor bearing rats for four days by means of total parenteral nutritional methods to investigate the interaction of caloric intake and the development of fatty liver. Compared with the total parenteral nutrition control group the triglyceride content of the liver rose significantly in the group given valine-depleted amino acid imbalance solution with an increase in caloric intake. Plasma total protein and albumin significantly decreased. The very-low-density lipoprotein concentration in serum was also significantly lower than that in the control group. Valine-depleted amino acid imbalance caused hypoproteinemia, suggesting a fall in synthesis of apolipoproteins in the liver indispensable for lipid release. Along with the increase in the total caloric intake, triglyceride synthesis in the liver increased, resulting in augmentation of fatty content of the liver, probably because of the decreased lipid release. PMID- 9200159 TI - The practitioner's dilemma: ethics, cost constraint, and quality of care issues in managed care. PMID- 9200160 TI - Role of optometric vision therapy for surgically treated strabismus patients. AB - BACKGROUND: Occasionally, co-management involving both optometry and ophthalmology is needed to optimize treatment outcome for the strabismic patient. METHODS: JB, a 47-month-old consecutive esotrope presented to our clinic. Two previous attempts to surgically correct her exotropia had failed and the parents sought another treatment approach. We recommended optometric vision therapy (VT) to improve sensorimotor fusion before any further surgery. After 31 VT sessions (bi-weekly for a time, then weekly), before a third scheduled surgery, sensorimotor fusion was good in the amblyoscope, but unstable with neutralizing prism in free-space. We recommended surgery be postponed, but the family proceeded. Esotropia recurred with constant suppression. After additional VT, JB developed stable sensorimotor fusion and random dot stereopsis in free-space with neutralizing prism. A fourth surgery was then performed resulting in esophoria at all distances with good sensory fusion. RESULTS: Twenty-one months postoperatively, JB remains nonstrabismic with good sensory fusion. CONCLUSIONS: Clinicians should understand the roles and limitations of available treatment options. Surgery reduces the magnitude of the deviation, whereas optometric VT provides the unique role of establishing normal sensory processing. PMID- 9200161 TI - Biosocial consequences of poverty: associated visual problems. AB - Although the impact of poverty on the health and development of children is readily acknowledged, the extent of accompanying visual functional and perceptual motor disorders has not received very much attention. These visual disorders are shown to be linked with poverty. In particular, research studies to support the notion that neurointegrative and concomitant visual problems can be the result of malnutrition, low birthweight, teenage pregnancy, and maternal complications of pregnancy are cited and discussed. The association between perceptuocognitive functioning in children and sociodemographic factors are examined and related to central nervous system maturation in a hostile environment. The role of the optometrist in diagnosing and treating children with visual and neurointegrative problems is reviewed. Special emphasis is placed on the potential for optometric intervention to improve the ability of disadvantaged children to respond more effectively to classroom learning and enable them to make fuller use of the intelligence they possess. The authors recommend that optometrists play a vital role in their communities to ensure the rendering of appropriate professional treatment for economically impoverished and socially disadvantaged children. PMID- 9200162 TI - Protein changes during aging and the effects of long-term cortisol treatment in macaque monkey lens. AB - Macaca nemestrina pig-tail macaques were administered daily oral doses of 3.85 or 5.78 mg/kg of cortisol for 1 year. The ages of the macaques were from 19 to 29 years. After 1 year, lenses were observed using a slitlamp ophthalmoscope and the stage of cataract was classified in each eye. Enucleated lenses were analyzed for content of soluble and insoluble proteins. Lens proteins were analyzed using SDS polyacrylamide gel electrophoresis (SDS-PAGE) and the changes in lens proteins were quantified using densitometry of the individual gels. Untreated control lenses were obtained over the range of 4 to 29 years of age and the proteins were analyzed. A slow progressive increase in the cataract stage and in the proportion of insoluble protein aggregates corresponded with the animal age, not the cortisol treatment. The observed changes in the protein components may suggest an important role for cytoskeletal proteins in lens transparency during aging. Exposure to high doses of oral steroids over a period of 1 year did not result in detectable modification of crystallin or cytoskeletal proteins. Even at high doses, longer exposure may be necessary to produce the cataract associated with steroid administration. PMID- 9200163 TI - Changes in lactate dehydrogenase activity in bovine corneal stroma and epithelium in response to in vitro toxic challenges in the enucleated eye test. AB - PURPOSE: To assess whether changes in lactate dehydrogenase (LDH) activity, as a marker of cell damage, could be detected in corneal stroma and corneal epithelium after toxicity measurements using the enucleated eye test (EET). METHODS: The corneal surface of isolated bovine eyes was continuously wetted with commercial balanced salt solution (BSS) over 4 h at 37 degrees C and central corneal thickness (CCT) was measured. Initial 5-min challenges were made with dilute solutions of benzalkonium chloride (up to 0.032%) or NaOH (up to 0.4 M). The residual LDH activity in the corneal epithelium and corneal stroma were then assessed. RESULTS: Dose-dependent increases in CCT of up to 70% were measured. A strong correlation (r = -0.961) was found between increases in CCT and decreases in stromal LDH activity, but the correlation was much less (r = -0.512) for epithelial LDH activity. CONCLUSIONS: LDH activity can be used as one marker for cell integrity in the corneal stroma and epithelium, although dose-dependent correlations may not be high. PMID- 9200164 TI - Sequential staining: the effects of sodium fluorescein, osmolarity, and pH on human corneal epithelium. AB - BACKGROUND: Previous reports have suggested that sequential applications of sodium fluorescein (NaF) to the ocular surface cause loss of epithelial cells. In those experiments the solutions were free of preservatives, but delivered a hypertonic and alkaline load. It is possible that either the hyperosmolarity, NaF, and/or alkalinity may have contributed to the epithelial cell loss. Our study explored the possible impact of these three factors on epithelial integrity. METHODS: We used a paradigm in which we designed four test solutions to isolate the cytotoxic effects of the three factors. Fifteen subjects were exposed to one of the solutions on separate visits. One solution was instilled in both eyes, every 3 min, for a total of seven applications, and a slitlamp examination was performed. Staining was graded on a scale of 0 to 3 for each of five corneal sectors. RESULTS: Application of solution 1 (NaF, hyperosmotic, and alkaline) resulted in staining for all subjects. For the other three solutions (without NaF), insignificant staining occurred. CONCLUSIONS: The epithelium tolerates changes in pH and osmolarity between 7.2 to 7.8 and 290 to 350, respectively. However, NaF when applied in multiple doses may be cytotoxic. PMID- 9200165 TI - Optical performance of aspheric concave ophthalmic lenses: the effect of vertex distance. AB - Variations of power errors in the periphery of concave aspheric lenses were assessed as a function of vertex distance. Comparisons were made between the performance of different types of spherical lenses and that of aspheric lenses ( 6.00 and -8.00 D lenses were used). Off-axis measurements were made by means of a modified focimeter, which enabled rotation of the lens around the center of the vertex sphere. Reduction of the vertex sphere's radius from 27 to 23 mm simulated a 4-mm decrease in vertex distance. Results of the measurements are compared with a computed model. These results indicate that off-axis performance is more affected by a reduction of vertex distance in the case of aspheric lenses compared to spherical lenses. Resulting increases in oblique astigmatism (OA) and mean oblique error (MOE) are at the limit of clinical acceptance. The importance, for the practitioner, of respecting the vertex distance selected for a specific aspheric design when fitting this type of lens is emphasized. PMID- 9200166 TI - Long wavelength pass filters designed for the management of color vision deficiencies. AB - This study reports on the effectiveness of long wavelength pass filters dispensed as tinted spectacles as an aid for individuals with congenital red-green color vision deficiencies. The effectiveness of the filters was evaluated by the performance on a series of clinical color vision tests and a questionnaire after the subjects had tried the lenses for 1 week. The lenses improved performance on color vision tests that required discrimination between large color differences, particularly between red and green hues. However, performance was degraded on tests which required fine color discrimination or used colors that were located parallel to the tritan confusion axis. The improved performance on certain tests was primarily based on brightness artificats induced by the filters, whereas the degraded performance on the other tests was due to the absorption of short- to midwavelength light by the filters. A slight majority (56%) of the subjects rated the filters as being moderate to highly effective in improving their color discrimination. Nevertheless, only 17% were interested in actually purchasing a pair. Common reasons for rejecting the filters were the color distortions produced by the red filters and fewer colors were actually perceived when wearing the filters. PMID- 9200167 TI - Performance of a color indicator in a disinfecting solution for the maintenance of soft contact lenses. AB - An experimental study of Allergan's Oxysept Comfort system was performed by measuring the slight reddish hue that appears in the disinfecting solution, indicating to the users that their lenses are again ready to be worn. The temporal evolution of the color of the solution has been measured under standardized conditions and analyzed in the CIELAB system, from the perspective of the typical threshold discrimination of the human eye. Color differences between neutralized and non-neutralized solutions occurred in an appropriate direction of the color space to enhance discrimination and were clearly perceptible by normal observers (greater than 9.7 +/- 1.2 CIELAB units). Colorimetric analyses have been used to draw conclusions regarding observers with defective color vision. The color of the solution changes abruptly, approximately 25 min after the neutralization process begins, and remains nearly constant after about 60 min, this agreeing well with the temporal evolution of the hydrogen peroxide concentration. PMID- 9200168 TI - Failure of convergence of two lines of research: buried treasure. PMID- 9200169 TI - An assessment of the efficacy of physical therapy and physical modalities for the control of chronic musculoskeletal pain. AB - An analysis of review articles and controlled clinical trials for temporomandibular disorders and similar chronic musculoskeletal pain disorders was carried out. Although little evidence was found that any specific therapy had long-term efficacy greater than placebo, we did find strong evidence that symptoms improve during treatment with most forms of physical therapy, including placebo. When the frequency of significant between-group differences in trials that used placebo and waiting list control (i.e., no treatment) groups were compared, it was found that treatment was better than placebo in only 7/22 trials, whereas treatment was almost always better than no treatment (15/16). This difference was highly significant (P = 0.001). A similar analysis of trials that included more than one treatment group showed that while equal amounts of treatment were usually associated with equal outcome (9/10), unequal treatment regimes led to unequal outcome (10/15; P = 0.012). The group that received the most therapy appeared to do best. In conclusion, it seems that patients are helped during the period that they are being treated with most forms of physical therapy. However, most of these therapies have not been shown to be more efficacious than placebo. PMID- 9200171 TI - Thalamic NMDA receptors modulate inflammation-produced hyperalgesia in the rat. AB - The effects of inhibition of thalamic NMDA receptor function and synthesis on thermal and mechanical hyperalgesia induced by hindlimb intraplantar injection of carrageenan in the rat were studied in the 'acute' phase (within 3-5 h) and the 'subacute' phase (24 h) after carrageenan administration. Blockade of NMDA receptors was produced by intrathalamic injection of D,2-amino-5-phosphonovaleric acid (D-APV) and NMDA receptor synthesis was decreased (or not) by pretreatment of rats with intrathalamic (hindlimb representation area) injections of antisense, sense or missense oligodeoxynucleotides (ODNs) directed against the NR1 subunit of the NMDA receptor complex. Treatment with D-APV, but not saline, in the contralateral (but not ipsilateral) thalamus significantly reduced both the acute thermal and mechanical hyperalgesia in the injected paw; these same rats demonstrated significantly less thermal and mechanical hyperalgesia in the sub-acute phase than rats that had received saline or D-APV in the ipsilateral thalamus. None of the treatments had any effect on withdrawal responses of the uninjected hindpaw. Rats pretreated with NR1 sense or missense ODNs developed both thermal and mechanical hyperalgesia that was equivalent in magnitude and duration to rats that received intrathalamic saline injections. In contrast, rats pretreated with NR1 antisense ODN did not develop either acute or subacute thermal hyperalgesia; they developed less mechanical hyperalgesia than saline, sense or missense ODN-treated rats. Antisense ODN-treated rats also displayed a decrease in the number of thalamic NMDA receptors as determined by receptor binding assay. These results suggest an involvement of thalamic NMDA receptors in the development and maintenance of hyperalgesia associated with neurogenic inflammation in a model of tonic pain. PMID- 9200170 TI - Benzodiazepine mediated antagonism of opioid analgesia. AB - Activation of supraspinal gamma-aminobutyric acid-A (GABAA) receptors is known to result in antagonism of opioid analgesia. Since benzodiazepines enhance the action of GABA at GABAA receptors, we hypothesized that administration of these agents for preoperative sedation might antagonize the analgesic effects of opioids administered postoperatively. If so, then administration of the benzodiazepine antagonist flumazenil should enhance postoperative morphine analgesia. In a double-blind, placebo-controlled study of patients who received a preoperatively administered benzodiazepine (diazepam) for sedation and a postoperatively administered opioid (morphine) for analgesia, we investigated opioid-benzodiazepine interactions affecting postoperative dental pain. We found that flumazenil significantly enhanced morphine analgesia consistent with the hypothesis that the preoperatively administered benzodiazepine exerts an ongoing antianalgesic effect. In addition, we followed these patients over the first and second postoperative days to determine if there were differences between the drug groups in post-discharge pain, analgesic consumption, or side-effects. Participants receiving flumazenil reported significantly less post-discharge nausea and used significantly less ibuprofen. Since post-discharge pain levels were not significantly different, these results suggest that the patients receiving flumazenil required less analgesic medication to achieve a comparable level of pain control. In summary, our results indicate that the benzodiazepine antagonist flumazenil enhances morphine analgesia and decreases post-discharge side-effects as well as post-discharge need for analgesic medication. PMID- 9200172 TI - Hydromorphone analgesia after intravenous bolus administration. AB - This study investigated the analgesic effects of three intravenous bolus doses of hydromorphone (10, 20, 40 micrograms/kg) on experimental pain measures in normal humans. Ten healthy male volunteers participated in four study sessions, one for each of the hydromorphone doses as well as a placebo (saline). They received the four treatments in counterbalanced order under double-blind conditions and with study days at least 1 week apart. During each session subjects underwent repeated electrical tooth pulp stimulation at intensities sufficient to elicit a rating of 'strong pain' before drug administration. Subjective pain reports (PRs) and dental evoked potential amplitude measures (EPs) served as analgesic effect indicators. We observed dose-dependent analgesia as measured by both PR (P = 0.009) and EP (P = 0.017). Area under the PR versus time curve as well as the EP versus time curve decreased in a log dose-dependent fashion. Although the peak effect was poorly defined, the onset of analgesia was rapid, within 5 min, and maximum analgesic effect was seen between 10 and 20 min after maximum plasma hydromorphone concentration. However, within sessions we found a poor correspondence between hydromorphone plasma concentration and effect. Compared to pain report data from other human studies done in our laboratory, hydromorphone has a shorter time to peak effect compared to morphine, and overall, hydromorphone hydrochloride is approximately five times as potent as morphine sulfate on a milligram basis. PMID- 9200173 TI - Cutaneous field stimulation (CFS): a new powerful method to combat itch. AB - Scratching the skin, while instantly relieving itch, often aggravates itch over time due to skin injury. To relieve itch, without damaging the skin, a new technique termed cutaneous field stimulation (CFS) was developed and tested on 21 subjects. CFS uses a flexible plate with needle-like electrodes (n = 16) to electrically stimulate nerve fibres in the superficial skin. The electrodes were stimulated consecutively (4 Hz per electrode, pulse duration 1 ms, intensity 0.4 0.8 mA, 25 min). CFS resulted in a pricking and burning sensation that usually faded rather quickly. The burning sensation was still present during a selective block of impulse conduction in myelinated fibres indicating that nociceptive C fibres are activated by CFS. Furthermore, a flare reaction developed around the CFS electrodes indicating activation of axon reflexes in nociceptive C-fibres. Itch, elicited by transdermal iontophoresis of histamine, was abolished within the skin area pre-treated with CFS, and was reduced to 14% of control 10 cm distally. Contralateral effects were small or non-existent. After 4 h, itch was reduced ipsilaterally to 32% of control. In comparison, 2 h after transcutaneous electrical nerve stimulation (TENS; 10-20 mA, 100 Hz, 25 min) ipsilateral itch was reduced to 56% of control. In conclusion, CFS offers a powerful new method for combating itch. It is suggested that CFS acts through endogenous central inhibitory mechanisms that are normally activated by scratching the skin. PMID- 9200174 TI - Differential activities of intrathecal MK-801 or morphine to alter responses to thermal and mechanical stimuli in normal or nerve-injured rats. AB - Nerve ligation injury in rats results in reduced nociceptive and non-nociceptive thresholds, similar to some aspects of clinical conditions of neuropathic pain. Since underlying mechanisms of hyperalgesia and allodynia may differ, the present study investigated the pharmacology of morphine and MK-801 in rats subjected to a tight ligation of the L5 and L6 nerve roots or to a sham-operation procedure. Response to acute nociception was measured by (a) withdrawal of a hindpaw from a radiant heat source, (b) withdrawal of the tail from a radiant heat source or (c) the latency to a rapid flick of the tail following immersion in water at different noxious temperatures. Mechanical thresholds were determined by measuring response threshold to probing the hindpaw with von Frey filaments. Nerve ligation produced a significant, stable and long-lasting decrease in threshold to mechanical stimulation (i.e., tactile allodynia) when compared to sham-operated controls. Standardization of the diameter of the filaments (to that of the largest filament) did not alter the response threshold in nerve-injured animals. Nerve ligation produced decreased response latency of the ipsilateral paw (i.e., hyperalgesia) when compared to that of sham-operated rats. Tail-flick latencies to thermal stimuli induced by water at constant temperatures (48 degrees, 52 degrees or 55 degrees C) or by radiant heat were not significantly different between nerve-injured and sham-operated groups. At doses which were not behaviorally toxic, MK-801 had no effect on tactile allodynia. At these doses, MK 801 blocked decreased paw withdrawal latency to radiant heat in nerve-injured rats, but did not significantly elevate the response threshold of sham-operated rats. Systemic (i.p.) or intracerebroventricular (i.c.v.) doses of morphine previously shown to be antiallodynic in nerve-ligated rats did not affect the response to probing with von Frey filaments in sham-operated controls. Intrathecal (i.t.) morphine did not change paw withdrawal thresholds elicited by von Frey filaments of either nerve-ligated rats (as previously reported) or of sham-operated rats at doses maximally effective against thermal stimuli applied to the tail or foot. Spinal morphine produced dose-dependent antinociception in both nerve-injured and sham-operated groups in the foot-flick test but was less potent in the nerve-injured group. Presuppression of hyperalgesia of the foot with i.t. MK-801 in nerve-injured animals did not alter the potency of i.t. morphine. I.t. morphine was also active in the tail-flick tests with decreased potency in nerve-injured animals and, at some stimulus intensities, with a decreased efficacy as well. These data emphasize the distinction between the inactivity of morphine to suppress mechanical withdrawal thresholds (as elicited by von Frey filaments) and the activity of this compound to block the response to an acute thermal nociceptive stimulus in sham-operated or nerve-injured rats. It appears that nerve ligation injury produces a thermal allodynia/hyperalgesia which is likely dependent upon opioid-sensitive small-diameter primary afferent fibers and a mechanical allodynia which may be largely independent of small-fiber input. PMID- 9200175 TI - Modulation of cutaneous nociceptor activity by electrical stimulation in the brain stem does not inhibit the nociceptive excitation of dorsal horn neurons. AB - In anesthetized cats, recordings were obtained from single lumbar dorsal horn neurons and from primary afferent fibers of the posterior tibial nerve excited by controlled noxious radiant heating of glabrous hindpaw skin. Electrical stimulation in four brain stem regions (periaqueductal gray and lateral reticular formation in the midbrain, raphe and reticular formation in the medulla) during noxious skin heating markedly reduced the nociceptive excitation of the dorsal horn neurons. In contrast, such brain stem stimulation had small and variable effects upon the noxious heat-evoked activity in the primary afferent fibers; both increases and decreases were observed. The brain stimulation also produced transient changes in blood pressure, suggesting that circulatory effects may underlie the mechanism of nociceptor modulation. It is concluded that brain stem stimulation can modulate cutaneous nociceptor activity, but that this modulatory effect on nociceptor inflow is too small and inconsistent to explain the marked descending inhibition of the nociceptive excitation of dorsal horn neurons. PMID- 9200176 TI - Cholecystokinin antisense RNA increases the analgesic effect induced by electroacupuncture or low dose morphine: conversion of low responder rats into high responders. AB - The analgesic effects of the rat in response to electroacupuncture (EA) or low dose morphine (3 mg/kg) show marked individual variations. In the midbrain periaqueductal gray (PAG) of the rat, the content of the neuropeptide cholecystokinin octapeptide (CCK-8) was found to be significantly higher in the low responder (LR) rats as compared to that in the high responders (HR). Since PAG has been shown to be a strategic site for CCK-8 to exert an anti-opioid action, a high CCK content in PAG may account for the low analgesic responsiveness to EA and morphine. In order to block the expression of the gene encoding preproCCK in the brain, antisense CCK expression vector pSV2-CCKAS was microinjected into the lateral cerebral ventricle of the rat, leading to a decrease of the CCK-mRNA as well as the CCK-8 content in rat brain. This effect started 4 days after the intracerebroventricular (i.c.v.) injection of the antisense expression vector, and lasted no more than 1 week. This procedure was shown to be very effective in converting LR rats into HR for EA analgesia and morphine analgesia, and also delayed the development of tolerance elicited by prolonged EA stimulation or repeated morphine administration. The time course of the augmentation of opioid analgesia (4-6 days after the i.c.v. injection of the expression vector) paralleled the decrease of the brain CCK-8 content. The results argue that blocking the CCK gene expression in the brain may tilt the balance between opioid and anti-opioid peptides in favor of the former, thus strengthening the EA analgesia and morphine analgesia, and delaying the development of opioid tolerance. PMID- 9200177 TI - Inhibition of the blink reflex R2 component after supraorbital and index finger stimulations is reduced in cluster headache: an indication for both segmental and suprasegmental dysfunction? AB - Peripheral as well as central mechanisms are thought to play a role in cluster headache pathogenesis. We have studied recovery curves of the R2 component of the blink reflex after conditioning by supraorbital or index finger stimuli in 10 episodic cluster headache (CH) patients during a cluster period and in 10 healthy controls. There was no significant change of R2 threshold, latency or area in CH patients. After paired supraorbital stimuli, R2 recovered more rapidly in patients on the symptomatic side. After index stimulations, R2 recovery was more rapid on both symptomatic and non-symptomatic sides in patients compared to controls. Naloxone (0.4 mg) i.v. in two subjects partially reversed the R2 suppression induced by index finger stimuli. The unilateral decrease of R2 inhibition after a segmental supraorbital stimulus most likely reflects sensitisation in the spinal trigeminal nucleus. Whether the latter is due to irritation of the ophthalmic nerve within the cavernous sinus region, which is thought to be pivotal in CH pathogenesis, remains to be proven. In addition, we propose that the bilateral deficit of R2 inhibition after an extrasegmental exteroceptive stimulus might reflect hypoactivity of reticular nuclei, possibly because of reduced central opioid activity. PMID- 9200178 TI - Effects of the bradykinin B1 receptor antagonist des-Arg9[Leu8]bradykinin and genetic disruption of the B2 receptor on nociception in rats and mice. AB - The contributions of B1 and B2 bradykinin receptors to acute and chronic inflammatory hyperalgesia were examined using the peptide B1 receptor antagonist des-Arg9[Leu8]bradykinin and transgenic Bk2r-/- mice. In normal rats and mice, des-Arg9[Leu8]bradykinin (30 nmol/kg i.v. or s.c.) inhibited carrageenan-induced hyperalgesia and the late phase nociceptive response to formalin. The active dose range was narrow, suggesting partial agonist activity of this peptide. In rats with monoarthritis, des-Arg9[Leu8]bradykinin (up to 30 nmol/kg i.v.) failed to reduce the number of vocalisations elicited by gentle flexion and extension of the inflamed limb; however, hyperalgesia was exacerbated by administration of the B1 receptor agonist des-[Arg9]bradykinin (100 nmol/kg i.v.), consistent with other evidence for local induction of B1 receptors during adjuvant-induced arthritis. The nociceptive response to intraplantar injection of bradykinin (10 nmol) and hyperalgesia induced by carrageenan (0.6 mg) were absent in Bk2r-/- mice, indicating that stimulation of B2 receptors is an essential step in the initiation of some nociceptive and inflammatory reactions. However, the nociceptive response to formalin (2.5% intraplantar), including inhibition of the late phase by des-Arg9[Leu8]bradykinin (0.3 nmol), and induction of thermal hyperalgesia by Freund's adjuvant (0.1%) appeared intact in Bk2r-/- mice. These findings support other evidence for an involvement of B1 receptors in inflammatory hyperalgesia and suggest that B1 receptor antagonists may be clinically useful as anti-inflammatory and analgesic drugs. PMID- 9200179 TI - Chronic treatment with systemic morphine induced tolerance to the systemic and peripheral antinociceptive effects of morphine on both carrageenin induced mechanical hyperalgesia and spinal c-Fos expression in awake rats. AB - The effects of intravenous (3 mg/kg i.v.) and intraplantar (50 micrograms/50 microliters i.pl.) morphine were investigated on spinal c-Fos expression induced 2 h after intraplantar carrageenin (6 mg/150 microliters of saline) and on carrageenin (2 mg/150 microliters of saline) induced mechanical hyperalgesia, at day 4, in both naive and chronic morphine treated (80 mg/kg/day s.c. on days 1, 2 and 3) rats. In naive rats, i.v. and i.pl. morphine significantly decreased spinal c-Fos expression (64 +/- 4% and 44 +/- 4% reduction of control carrageenin c-Fos expression, P < 0.0001 for both, respectively) and mechanical hyperalgesia (maximal increase: 326 +/- 29%, P < 0.0001 and 87 +/- 5%, P < 0.005 of control carrageenin paw pressure vocalisation threshold (VTPP), respectively), which only developed in the carrageenin injected paw. Both treatments were ineffective in chronic morphine treated rats (92 +/- 9% and 106 +/- 6% of control carrageenin c Fos expression; 33 +/- 17% and 30 +/- 15% increase of control carrageenin VTPP, respectively). Furthermore, only i.v. morphine increased the VTPP in the contralateral paw, in naive rats (maximal increase: 90 +/- 8%, P < 0.0001 of control carrageenin VTPP), its effects being significantly less pronounced than for the inflamed paw (P < 0.0001). These studies based on spinal c-Fos expression as an indirect marker of spinal nociceptive processes and on behavioural experiments clearly revealed that chronic treatment with systemic morphine induced tolerance to both its systemic and peripheral effects. PMID- 9200180 TI - A 'leaking' Synchromed pump. PMID- 9200181 TI - Respiratory depression following oral tramadol in a patient with impaired renal function. PMID- 9200182 TI - Comment on Ogon et al., PAIN, 64 (1996) 425-428. PMID- 9200183 TI - Response to Lariviere and Melzack, PAIN, 66 (1996) 271-277. PMID- 9200184 TI - Comments on Schwartz et al., PAIN, 65 (1996) 227-233. PMID- 9200185 TI - Mechanisms underlying arterial fragility and the complications of atherosclerosis. AB - The etiology of atherosclerosis must explain the development of primary pathological complications (intimal tears, ectasia, tortuosity, aneurysms and stenoses). They are interrelated and associated with destruction of mural architecture and concomitant loss of tensile strength (fragility) attributable to bioengineering fatigue. The complications become manifested clinically by ischemia, hemorrhage and pressure effects developing with greater frequency in association with hypertension, arteriovenous shunts or connective tissue disorders. Moreover they are produced experimentally and iatrogenically by hemodynamic means but are unexplained by other current etiological hypotheses. PMID- 9200186 TI - Expression of TA1, a rat oncofetal cDNA with homology to transport-associated genes, in carbon-tetrachloride-induced liver injury. AB - TA1, a novel rat oncofetal cDNA, is the predicted homolog of the human lymphocyte activation gene E16. The encoded peptides share high homology with transport associated and uncharacterized sequences in distant species, suggesting an important and conserved function in cellular homeostasis. Moderate steady-state levels of TA1 RNA were induced following acute and chronic CCl4-mediated liver injury. TA1 expression was either greatly reduced or absent in livers of animals receiving injury-protective doses of vitamin E in conjunction with CCl4. In contrast to the in vivo data, acute in vitro exposure of hepatocytes to CCl4 did not induce TA1 RNA. Our results indicate that TA1 is spatially and temporally associated with liver injury in vivo and may play an adaptive role in the hepatic response to environmental toxicants. PMID- 9200188 TI - Esophageal carcinogenesis in the rat: zinc deficiency and alcohol effects on tumor induction. AB - Sprague-Dawley male rats were fed zinc-deficient or supplemented diets for 2 weeks, administered a carcinogenic dose of methylbenzylnitrosamine and observed over 20 or more weeks for effects of superimposing excess zinc or alcohol on development of esophageal tumors. In three separate experiments it was shown that (1) excess zinc offered no protection, (2) switching diets during or after carcinogen exposure pointed toward involvement of zinc in both initiation and promotion, (3) neither ethanol nor 3-methyl butanol alone affected tumorigenesis but the two combined and superimposed on a zinc deficiency resulted in a significant enhancement of neoplasia. In one group of rats fed the zinc-deficient diet only, with no carcinogen, 4 rats developed neoplasms, one of which was malignant. Cell proliferation, an integral component of zinc deficiency, appears to be an important contribution to tumor induction in this model. PMID- 9200187 TI - Immunization with meningococcal membrane-bound lipooligosaccharide accelerates granulocyte recovery and enhances lymphocyte proliferation in myelosuppressed mice. AB - Protective effects of detergent-treated outer membrane vesicles (D-OMVs) prepared from the parent group B M986 strain and the nonencapsulated M986-NCV mutant in myelosuppressed mice were investigated in models of experimental septic shock. The effects of D-OMVs on expansion of the myeloid compartment, on spleen cell proliferation to mitogen stimulation, and on cytokines induced during this period were investigated. On 3 consecutive days, mice were injected with 1 microgram/kg of lipooligosaccharide (LOS) or lipopolysaccharide, or 75 micrograms/kg D-OMV followed by a single dose of cyclophosphamide (200 mg/kg) 24 h later. Eight weeks after the last injection, animals were challenged with a combination of galactosamine (400 mg/kg) and live Neisseria meningitidis. More than 90% of control mice died within 24 h when challenged with 10(5) CFU of bacteria. Mice immunized with LOS or D-OMV were rendered neutropenic but were protected against bacterial challenge of at least 10(7) CFU. At different time intervals, peripheral blood samples were obtained to characterize changes in circulating blood cells. The rise in absolute granulocyte numbers occurred 24 h earlier with peak cell counts about 3 times higher than those seen in the placebo groups. Peripheral blood cells from D-OMV-treated animals expressed about twofold more Gr 1 antigen (myeloid surface cell marker) than placebo-treated controls. The proliferative responses to both B and T cells were reduced in all treatment groups due to the effects of cyclophosphamide. D-OMV treatment afforded the greatest protection for mitogen-activated lymphocytes from the lethal effects of cyclophosphamide and also enhanced T and B cell proliferation. Low IL-1 beta levels and increases in serum IL-6 were detected in all treatment groups. In contrast, significant IFN-gamma and IL-3 levels were only detected in D-OMV treated groups. These results indicate that D-OMVs, which have reduced toxicity, have prophylactic potential in inducing specific cytokines that accelerate granulocyte recovery following cytoreductive therapy by promoting both proliferation and maturation of myeloid precursors as well as augmenting the immune system. PMID- 9200190 TI - Stealth virus epidemic in the Mohave Valley. I. Initial report of virus isolation. AB - Increasing numbers of patients within the Mohave Valley region of the United States are reporting symptoms attributable to atypical neurological illness. Many of these patients have experienced an acute-onset gastrointestinal illness during the spring and summer of 1996. Stealth viral cultures performed on the blood of 40 of these patients have been uniformly positive, yielding unequivocal transmissible cytopathic effect (CPE) in both human- and monkey-derived cell lines. One patient has died from a stealth-CPE-positive glioblastoma, while another patient has developed a plemorphic adenoma of the parotid. Viral cultures and epidemiological data support human-to-human, and probable human-to-dog, transmission of the Mohave stealth virus infection. PMID- 9200189 TI - Common variable immunodeficiency associated with myelocathexis and altered membrane sodium-proton antiport. AB - Alterations in protein kinase C (PKC) activity have been implied in the pathogenesis of common variable immunodeficiency (CVID). We analyzed amiloride sensitive red blood cell Na+/H+ exchange (sodium-proton antiport, SPA) and its response to protein kinase stimulation in a patient with CVID. Compared with healthy subjects or patients with sepsis, a unique pattern of SPA activation has been shown. The patient's SPA was decreased and unresponsive to PKC stimulation, whereas stimulation by insulin, a tyrosine kinase activator, restored SPA activity. An alteration of serine-threonine phosphorylation is suggested as a possible mechanism for the immune failure. PMID- 9200191 TI - Detection of RNA sequences in cultures of a stealth virus isolated from the cerebrospinal fluid of a health care worker with chronic fatigue syndrome. Case report. AB - A cytopathic stealth virus was cultured from the cerebrospinal fluid of a nurse with chronic fatigue syndrome. Reverse transcriptase-polymerase chain reaction (RT-PCR) performed on the patient's culture yielded positive results with primer sets based on sequences of a previously isolated African green monkey simian cytomegalovirus-derived stealth virus. The same primer sets did not yield PCR products when tested directly on DNA extracted from the cultures. The findings lend support to the possibility of replicative RNA forms of certain stealth viruses and have important implications concerning the choice of therapy in this type of patient. PMID- 9200192 TI - X-ray vision. PMID- 9200193 TI - Concerns about microabrasion. PMID- 9200194 TI - On Medicaid. PMID- 9200195 TI - Dentin bonding: SEM comparison of the dentin surface in primary and permanent teeth. AB - The literature suggest differences between primary and permanent teeth regarding the composition and morphology of the dentin. The purpose of this study was to compare the effect of two dentin conditioners on the micromorphology of the dentin surface of primary and permanent teeth. Human extracted and noncarious molars were divided into four groups and conditioned with either 10% phosphoric acid (All-Bond 2) or 10% maleic acid (Scotchbond Multi-Purpose) for different time periods. SEM photomicrographs (1500x) were taken from the conditioned dentin and evaluated blindly by three calibrated examiners. The results indicate that the smear layer was removed more easily from primary teeth than from permanent teeth (P = 0.0001), which suggests greater reactivity to acidic dentin conditioners. We also found that the longer the time of application of dentin conditioner the more smear layer is removed (P = 0.0094). In comparing primary and permanent dentin, the results of this study indicate that less time is required for appropriate acid conditioning of primary dentin surfaces. Such a differentiated protocol for bonding to primary tooth dentin results in surface morphological characteristics similar to those found in conditioned permanent teeth. PMID- 9200197 TI - Comparison of air abrasion versus acid etch sealant techniques: six-month retention. AB - This study compared two techniques for placing sealant in an elementary school setting. Eighty-five children in grades 1 to 4 in two schools were assigned randomly to two sealant treatment groups: 1) acid etch technique (AE), and 2) air abrasion with no acid etch (KCP-1000). Noncarious, nonfilled occlusal, distolingual and buccal pit surfaces of first permanent molars were sealed. A total of 300 teeth received sealants, and 230 were evaluated at 6 months. Rates of complete sealant retention at 6 months were: occlusal surfaces, 97% for AE and 96% for KCP-1000; distolingual surfaces, 82% for AE and 49% for KCP-1000; and buccal surfaces, 77% for AE and 7% for KCP-1000. Differences in complete retention at 6 months between AE and KCP-1000 were not significant for occlusal surfaces (P = 0.14) but were significant for buccal and distolingual surfaces (P < 0.0001). Results suggest that sealants placed with air abrasion have retention rates for occlusal surfaces similar to AE. More research is needed to identify factors contributing to low retention rates on other surfaces for KCP-1000. PMID- 9200196 TI - A comparison of three resin bonding agents to primary tooth dentin. AB - This study determined the shear bond strength of resin composites to primary dentin using three dentin adhesives and the presence or absence of a hybrid zone. The buccal and lingual surfaces of 40 recently extracted noncarious primary teeth were ground flat with SiC paper ending with the 600 grit. The teeth were divided at randomly into eight groups of five teeth (10 surfaces) each: 1) Unetched dentin, dry dentin, All-Bond 2/Bis-Fil P; 2) Unetched dentin, moist dentin, All Bond 2/Bis-Fil P; 3) Dentin etched for 15 sec with 10% phosphoric acid, dry dentin, All-Bond 2/Bis-Fil P; 4) Dentin etched for 15 sec with 10% phosphoric acid, moist dentin, All-Bond 2/Bis-Fil P; 5) Dentin etched with 10% maleic acid for 15 sec, dry dentin, Scotchbond Multi-Purpose/Z100; 6) Dentin etched with 10% maleic acid for 15 sec, moist dentin, Scotchbond Multi-Purpose/Z100; 7) Dentin etched with 10 citric acid/3% ferric chloride, dry dentin, Amalgambond Plus/Z100; 8) Dentin etched with 10 citric acid/3% ferric chloride, moist dentin, Amalgambond Plus/Z100. All teeth were thermocycled 1000x (5 and 55 degrees C, 30 sec dwell time), and shear bond strength testing was conducted using an Instron (crosshead speed 0.5 mm/min). Failure sites after debonding were examined with the SEM. For each group, one additional tooth was used to prepare two class V cavities (one facial and one lingual) restored according to the specification in each group, sectioned buccolingually and examined with the SEM. The results, in MPa, were: 1) 12.55 +/- 5.97; 2) 10.41 +/- 6.16; 3) 9.94 +/- 7.26; 4) 12.25 +/- 4.70; 5) 13.02 +/- 8.01; 6) 16.51 +/- 8.62; 7) 12.51 +/- 8.95; 8) 17.93 +/- 6.44. ANOVA and Student-Newman-Keuls tests showed no statistically significant differences. SEM evaluation showed that the smear layer was removed in all groups exposing primary dentin tubules infiltrated by resin. A resin-reinforced hybrid layer was readily seen in all specimens. PMID- 9200198 TI - Microleakage of cements for stainless steel crowns. AB - Microleakage is related to recurrent decay, inflammation of vital pulps, and reinfection of previously treated root canals. The purpose of this investigation was to compare the abilities of new adhesive cements and conventional nonadhesive controls to prevent microleakage under stainless steel crowns on primary anterior teeth. Standardized preparations were made, and stainless steel crowns were adapted. Specimens were assigned randomly to cement groups: zinc phosphate (ZP), polycarboxylate (PC), glass-ionomer (GI), resin-modified glass-ionomer (RMGI), RMGI with a dentin bonding agent (RMGI + DBA), adhesive composite resin (ACR) and zinc oxide eugenol (ZOE). Specimens were stored in water, aged artificially, stained, embedded, and sectioned, and the microleakage was measured. Group means and standard errors were calculated. ANOVA discerned differences among groups (P < 0.0001), and Turkey's multiple comparisons testing (P < 0.05) ranked the groups from least to most microleakage as follows: [RMGI + DBA, RMGI, ACR, GI], [ZP], and [PC, ZOE]. The adhesive cements significantly reduced microleakage. PMID- 9200199 TI - An in vitro comparison of four surface preparation techniques for veneering a compomer to stainless steel. AB - Compomers are a new class of materials reportedly having the anticariogenicity and the bonding ability to metals similar to glass ionomers while maintaining the high esthetic qualities of composite resins. The purpose of this study was to determine and evaluate the shear bond strength and fracture pattern of a compomer (Dyract) to stainless steel crowns (SSCs) using different mechanical and chemical retention procedures for possible future development of a chair-side technique of producing esthetic SSCs. Thirty-two Unitek SSCs, divided into four groups, were mounted in autopolymerizing acrylic resin so that the resulting specimen has the crown's flat lingual surface projecting above and parallel to the top surface of the acrylic resin block. Dyract was placed in transparent nylon cylinders (3 x 3 mm) and bonded to SSC's surfaces directly (group 1) or following sandblasting of the SSCs (group 2). In group 3, Dyract was bonded to stainless steel lingual cleats that were previously spot-welded to the SSCs. In group 4, Dyract was bonded to sandblasted SSC's surfaces using Scotchbond Multi-Purpose Plus dental adhesive. Specimens were placed in deionized water for 1 hr at 37 degrees C. Shear bond strength was measured using a universal testing machine. The mean (SD) shear bond strengths in MPa for groups 1-4 respectively were as follows: 2.998 (1.381), 9.518 (2.464), 13.909 (1.653), and 9.372 (3.723). One-way ANOVA and Tukey's multiple range tests revealed a statistically significant difference between the groups (P < 0.00001). While no significant difference was found between groups 2 and 4 in which Dyract-PSA prime/adhesive and Scotchbond Multi Purpose Plus dental adhesive were used, group 3 had significantly higher shear bond strength than other groups. Stereoscopic and SEM examinations revealed adhesive and mixed bond failures. It is concluded that the bond strength of Dyract to SSCs could be enhanced significantly by applying simple mechanical means of retention that could be available in dental offices. PMID- 9200200 TI - Composite rebonding to stainless steel metal using different bonding agents. AB - The purpose of this study was to determine the in vitro bond strengths of composite rebonded to stainless steel crown metal (SS) using five different bonding agents after composite to SS bond failure had been produced. The adhesive systems were applied to the failed bonds following the manufacturers' instructions and, as a control, composite was bonded to SS without using a bonding agent. Each group was then divided into two subgroups: mechanically prepared (MP), in which the SS was roughened by a diamond bur, and unprepared (NMP), in which no modification of the SS was done. ESPE VISIO-GEM composite was placed in a plastic mold and light cured to the treated SS. Samples were stored in water at 37 degrees C for 72 hr, thermocycled for 500 cycles between 5 and 55 degrees C, and mounted in an Instron Universal Testing Machine. Caulk's Adhesive System provided significantly higher rebond strength (228.97 +/- 106.9 kg/cm2) than the other materials, and mechanical surface preparation offered no significant advantages. PMID- 9200201 TI - Relationship of tonsil size on an airway blockage maneuver in children during sedation. AB - A previous report suggested that airway compromise without self-correction may occur in pediatric dental patients sedated with chloral hydrate (CH) and nitrous oxide (N2O) and may be interpreted as "deep" sedation. The purpose of this institutionally approved study was to determine 1) the association between the size of the tonsils and 2) the degree of expired carbon dioxide (CO2) and oxygen saturation (SaO2) changes to simulated airway obstruction using the Moore head tilt maneuver for 30 sec or less. Thirty healthy children (ASA I), aged 22-40 months, were evaluated for tonsil size and sedated with CH (50 mg/kg) and hydroxyzine (2 mg/kg) and supplemented with N2O. Pulse oximetry and capnography were used to monitor the child. During the restorative phase when the patient appeared asleep, the head was rolled forward with the chin touching the chest for a period of 30 sec. Changes in SaO2 and CO2 waveform were observed during this period. The results indicated that seven children who had enlarged tonsils had blocked airways (as determined by capnography) lasting approximately 15 sec. The remaining children did not have enlarged tonsils and continued to exchange air appropriately. O2 levels did not change during this period. The results suggest that the likelihood of airway blockage increases with enlarged tonsils. In children without airway blockage, ventilation occurs unimpeded, and attempts to readjust the head may not occur. The association between airway blockage and patient responsiveness is discussed in relation to sedation levels. PMID- 9200202 TI - Use of endosseous implants in a 3-year-old child with ectodermal dysplasia: case report and 5-year follow-up. PMID- 9200203 TI - The efficacy of primer on sealant shear bond strength. PMID- 9200205 TI - Intraoral expansion screw replacement using a light-cured acrylic technique. PMID- 9200204 TI - Fluoridated and nonfluoridated unfilled sealants show similar shear strength. PMID- 9200206 TI - Cryosurgery in the management of mucoceles in children. PMID- 9200207 TI - Cell membrane-bound proteases: not "only" proteolysis. AB - Ectopeptidases are widely distributed among various cell systems. Their expression on an appropriate cell type is finely regulated, reflecting the specific functional cell implications and engagement in defined physiological pathways. Protein turnover, ontogeny, inflammation, tissue remodelling, cell migration and tumor invasion are among the many physiological and pathological events in which cell-surface proteases play a crucial role, both as effector as well as regulatory molecules. It has recently become clear that also non catalytic effects of membrane-bound proteases are of great importance in some biological regulations. They may generate specific signal transduction intracellularly, after reacting with certain target molecules. They may also play a pivotal role in cell-cell and cell-virus contact and recognition, as well as in binding to the extracellular matrix. This short review provides some insight into the multifunctional mechanisms attributed to cell membrane-bound proteases. PMID- 9200208 TI - Opposite effects of nitric oxide on identified inhibitory and excitatory cholinergic synapses of Aplysia californica. AB - The effects of nitric oxide on evoked acetylcholine (ACh) release were studied at two identified cholinergic neuro-neuronal synapses of the nervous system of the mollusc Aplysia californica. The NO-donor, 3-morpholinosydnonimine (SIN-1), decreased the amplitude of evoked inhibitory postsynaptic currents (buccal ganglion) and potentiated that of evoked excitatory postsynaptic currents (abdominal ganglion). SIN-1 acted by modulating the number of ACh quanta released. 8Br-cGMP mimicked the effects of NO on ACh release in both types of synapses thus pointing to the involvement of a NO-sensitive guanylate cyclase. Presynaptic voltage-dependent Ca2+ and K+ (IA and late outward rectifier) currents were not modified by SIN-1 suggesting another final target for NO/cGMP. The labelling of a NO-synthase by immunostaining in several neurones as well as the modulation of ACh release by L-arginine indicate that an endogenous NO synthase is involved in the modulation of synaptic efficacy in both buccal and abdominal ganglia. PMID- 9200209 TI - Production of hydrogen peroxide by alveolar macrophages from rats exposed to subacute and chronic hypoxia. AB - We have studied in vitro alveolar macrophages (AMs) obtained by tracheobronchial lavage from rats exposed to subacute (3 hours and 3 days) and chronic (3 weeks) hypoxia (FiO2 = 0.1) and from rats recovering from chronic hypoxia. Hydrogen peroxide production by AMs was measured by luminol-dependent chemiluminescence after AMs adhered to the walls of the measuring cuvette, after stimulation with phorbol-myristate-acetate (PMA), and when N-formyl-methionyl-leucyl-phenylanine (FMLP) was added subsequently to the cells which had been previously stimulated by adherence or PMA. H2O2 production after cell adherence and adherence combined with FMLP stimulation did not differ between the groups. The increase of H2O2 production after adding PMA, and FMLP in addition to PMA was significantly higher in AMs from rats exposed to hypoxia for 3 days than in the controls. Other experimental groups did not differ from their controls. It is concluded that 3 days' hypoxia primes AMs for enhanced production of H2O2 upon stimulation. The mechanism is probably at the level of synthesis of proteins involved in H2O2 production, or the shift to a more reactive phenotype of alveolar macrophages subpopulations. PMID- 9200210 TI - Transport of an antihypoxic drug stobadine across the blood-brain barrier in rat striatum and its influence on catecholamine-oxidative current: a voltammetric study under normal and anoxic/ischaemic conditions. AB - Differential pulse voltammetry with a carbon fibre microelectrode (ME) was used in pentobarbital-anaesthetized rats for monitoring the stobadine current (STB.C) on both sides of the blood-brain barrier (BBB) in the arterial bloodstream (BS) and in the corpus striatum (CS). The STB.C exhibited a distinct peak at a polarization voltage 540 +/- 30 mV (n = 4). The maximum of STB.C in BS attained 2 3 min after the STB administration (2.8 mg/100 g in 1.0 ml saline solution i.a.) was followed by a rapid decrease to about 20% within next 3 min. The STB readily passed across the BBB: the STB.C peak appeared in the CS in the 3rd minute and continued to rise up to the 30th min. The administration of STB did not prevent a large increase (1347 +/- 326%, n = 3) of the catechol-oxidative current (CA.OC) occurring in the CS between the 4th and 5th minute after cardiac arrest. However, a decrease of ME sensitivity to CA.OC in the presence of STB was observed. This fact leads to the speculation whether a similar "quenching" of dopamine by STB could not participate in the protective effects of STB observed in the brain exposed to hypoxia-reoxygenation. PMID- 9200211 TI - Inhibitory effect of ethanol on the Na(+)-ATPase activity of rat kidney proximal tubular cell plasma membranes. AB - The inhibitory effect of 2% ethanol (400 mM) in the incubation medium on several characteristics of the Na(+)-ATPase of basolateral plasma membranes from rat kidney proximal tubular cells was investigated. Ethanol did not change the Km of the enzyme for Mg2+, ATP or Na+; it did not change either the optimal pH or temperature values of the incubation medium for the enzyme to act and finally, it did not affect the apparent energy of activation of the enzyme. It was also found that 2% ethanol produced stronger inhibition of the ATPase when it is in an activated or stimulated state, than when it is working at its lower basal level. The presented results can be explained by assuming that 2% ethanol in the incubation medium inhibits Na(+)-ATPase activity by affecting the enzyme structure as well as its activating mechanism. PMID- 9200213 TI - The inhibition of angiotensin converting enzyme attenuates the effects of chronic hypoxia on pulmonary blood vessels in the rat. AB - The effect of chronic administration of angiotensin converting enzyme inhibitor on the development of hypoxic pulmonary hypertension was studied in rats. Male Wistar rats were-exposed for 3 weeks to isobaric hypoxia (10% O2) and treated with 10 mg/kg b.w. of Ramipril daily. The haemodynamic properties of the pulmonary vasculature were then measured in isolated blood-perfused lung preparation. Ramipril administration during the sojourn in hypoxia resulted in lower baseline perfusion pressure and lower slope of perfusion pressure-flow relationship compared to non-treated hypoxic rats. Partitioning of the distribution of pulmonary vascular resistance across the vascular bed by the occlusion technique showed that it was mainly due to a decrease of arterial and venous vascular resistances to blood flow. It is suggested that Ramipril attenuates the process of morphological reconstruction of pulmonary vasculature by chronic hypoxia rather than the level of vascular smooth muscle tone. PMID- 9200212 TI - Modulation of haemopoietic radiation response of mice by diclofenac in fractionated treatment. AB - The effects of diclofenac, an inhibitor of prostaglandin synthesis, were studied on the acute radiation syndrome elicited in mice by fractional irradiation. Several haematological parameters were evaluated in mice irradiated with 5x2 Gy and 3x, 4x, or 5x3 Gy (intervals between fractions 24 h) from a 60Co gamma-ray source. The animals were treated with diclofenac either before each fraction or only once before the last fraction. The survival of mice was recorded after the irradiation regimen of 5x3 Gy followed by a "top-up" dose of 3.5 Gy given 24 h after the last radiation fraction. Statistically significant enhancement of the endogenous spleen colony formation and of leukopoiesis was found in mice treated with diclofenac repeatedly, as compared with both saline-treated irradiated controls and animals administered a single diclofenac dose, if a sublethal total radiation dose had been accumulated. However, following accumulation of a lethal total radiation dose, slightly impaired survival was observed in mice given diclofenac. It follows from the results that diclofenac is a suitable drug for enhancing leukopoiesis impaired by sublethal fractionated irradiation. Nevertheless, undesirable side effects of this drug negatively influence the survival of experimental animals following a lethal accumulated radiation dose. PMID- 9200214 TI - Response of immobilized hepatocytes in a perfusion system to anoxia/reoxygenation: effect of cyclosporine A pretreatment. AB - The present study was designed to investigate the ameliorative effect of cyclosporine A (CsA) pretreatment on an anoxia/reoxygenation injury model by using immobilized perfused hepatocytes. Rats received an i.p. injection of two successive doses of CsA (5 mg/kg/day). Twenty-four hours later hepatocytes were isolated from CsA-treated and control rats. After hepatocyte isolation, immobilization, perfusion, induction of anoxia/reoxygenation, the structural and functional integrity of the hepatocytes was followed in a perfusion medium by measuring the leakage of lactate dehydrogenase (LD) and the time course of urea biosynthesis. CsA pretreatment reduced the initial rate of urea synthesis during normoxia but reduced the drop in the relative percentage rate of urea synthesis during the period of anoxia. LD leakage was increased threefold by anoxia and sevenfold by reoxygenation in cells of untreated animals. After CsA pretreatment in vivo, hepatocytes showed no increase in LD leakage into the medium. These findings demonstrate that the perfused immobilized hepatocytes can be used as a cellular model to assess the effects of liver insults such as anoxia/reoxygenation injury and that CsA modulates the injury. The mechanisms of CsA beneficial effects at the experimental level remain to be elucidated. PMID- 9200215 TI - Readiness potentials related to self-initiated movement and to movement preceded by time estimation: a comparison. AB - Two procedures for eliciting premovement potentials were compared: (1) the estimation of a 3 s interval elapsed after a warning auditory signal, and (2) classical "self-pacing". Eleven healthy right-handed subjects participated in the experiment, EEG records from scalp electrodes placed at CZ, C3+ and C4+ were analyzed. It has been shown that both procedures induced similar premovement potentials except that in the first procedure the early component of the potential was longer. The time estimation itself induced a negative slow potential consisting of a rapid set-up and a subsequent plateau. PMID- 9200216 TI - Lack of evidence for the interaction between renin-angiotensin-aldosterone system and endothelin in vivo. AB - Aim of the study was to reveal the possible factors regulating plasma endothelin (ET) levels in vivo in patients with essential hypertension (EH) by the simultaneous determination of plasma renin activity (PRA) and plasma aldosterone (ALD). In addition, the possible relationship between ET and circulating endothelial cells as a marker of endothelial damage was also investigated. The postural test revealed a significant increase of ET levels (26.7 +/- 9 vs 11.5 +/ 3 fmol/ml, p < 0.05) in the upright position. Captopril administration did not change plasma ET levels. No significant correlation was found between ET and PRA or ALD. Although a tendency to a positive correlation between ET and circulating endothelial cells (as the marker of endothelial perturbation) was found, it did not attain statistical significance. Our data do not support the suggestion that the renin-angiotensin-aldosterone system plays a major role in the regulation of ET secretion in vivo in EH. Postural stimulation of ET secretion may be caused by other factors than renin-angiotensin-aldosterone system. PMID- 9200217 TI - The action of pramiracetam on consequences of hypobaric hypoxia is only moderate. AB - The possible protective action of pramiracetam, a pyrrolidinone nootropic drug, against hypobaric hypoxia was studied in two age groups of immature rats with implanted electrodes. Epileptic afterdischarges induced by hippocampal stimulation were used as a measure of hypoxic damage. Pramiracetam did not substantially change these afterdischarges in 12- and 18-day-old rat pups which were not exposed to hypoxia. Hypobaric hypoxia (simulated altitude of 7000 m for one hour) led to prolongation of the first afterdischarge in both age groups. Pramiracetam did not influence this prolongation in 12-day-old rats. The first afterdischarge was shortened significantly in 18-day-old animals but not to the level of rats not exposed to hypoxia. The afterdischarges elicited by repeated stimulations (four times at 10 min intervals) did not differ in pramiracetam treated and control rats. PMID- 9200218 TI - Respiratory control index of mitochondria isolated from regenerating rat liver. AB - Mitochondria were isolated from regenerating rat liver 12, 24 and 48 h after partial hepatectomy. The "State 3" and "State 4" respiration were measured in the presence of succinate. The P/O quotient and respiratory control index (RCI) were calculated. The experimental data showed that the partial uncoupling of oxidative phosphorylation in regenerating liver mitochondria occurring in the early period of regeneration is partly due to free fatty acids. PMID- 9200220 TI - Cloning and sequence analysis of a novel insertion element from plasmids harbored by the carbofuran-degrading bacterium, Sphingomonas sp. CFO6. AB - Sphingomonas sp. CFO6 (a member of the alpha group of Proteobacteria) was isolated from a Washington soil by enrichment on the insecticide carbofuran as a sole source of carbon and energy. This strain has been shown to harbor five plasmids, at least some of which are required for catabolism of carbofuran. Rearrangements, deletions, and loss of individual plasmids resulting in the loss of the carbofuran-degrading phenotype were observed following treatment with heat or introduction of Tn5. Several putative insertion sequence elements of different sizes were cloned from these plasmids by trapping in pUCD800, a positive selection vector for isolation of transposable elements. Three of the most common putative IS elements (designated IS1412, IS1487, and IS1488) in the clone library were of different sizes and cross-hybridize with each other. An element hybridizing with IS1412, IS1487, and IS1488 was mobilized during growth of CFO6 at 42 degrees C and inserted into one of CFO6's plasmids (pCFO4), corresponding to a deletion in the plasmid and a loss of catabolic function. IS1412 was completely sequenced and its sequence analyzed. IS1412 is 1656 bp in length and possesses terminal partially matched inverted repeats of unequal length (17 and 18 bp). In addition, IS1412 contains an open reading frame which encodes a putative transposase with significant homology to the putative transposases of IS1380 from Acetobacter pasteurianus, HRS1 from Bradyrhizobium japonicum, and IS1247 from Xanthobacter autotrophicus. These related IS elements form part of a family of common IS elements distributed among members of the alpha group of the Proteobacteria. PMID- 9200219 TI - Identification of a chromosomal Shigella flexneri multi-antibiotic resistance locus which shares sequence and organizational similarity with the resistance region of the plasmid NR1. AB - The ampicillin resistance gene from Shigella flexneri 2a strain YSH6000 was cloned and shown by Southern hybridization analysis to be closely linked to the previously cloned streptomycin, chloramphenicol, and tetracycline resistance determinants, which are borne on a chromosomally integrated 99-kb element. Analysis of this chromosomal multi-antibiotic resistance locus revealed that it had a high level of sequence and organizational similarity to an equivalent region of the Shigella R-plasmid, NR1. However, the chromosomal locus exhibited several differences, including the presence of two stretches of sequence derived from IS elements, the precise insertion of a beta-lactamase encoding oxal cassette into the Tn21-borne integron In2, a possible 17.5-kb deletion, and the loss or inactivation of the mercury resistance determinant. Based on these data, it is proposed that the chromosomal locus arose following integration of an NR1 like plasmid. PMID- 9200221 TI - The hydrophobic TraM protein of pKM101 is required for conjugal transfer and sensitivity to donor-specific bacteriophage. AB - pKM101 is a self-transmissible plasmid of the IncN incompatibility group. Analysis of the DNA sequences of the genes required for conjugal transfer suggested the existence of a previously uncharacterized open reading frame, designated traM, that might be required for conjugation. Merodiploid strains containing transposon insertion mutations either in traM or in neighboring tra genes were used to demonstrate that traM constitutes a new complementation group essential for conjugation and donor phage sensitivity. The hydrophobicity profile of TraM suggests that it contains a signal sequence. The remainder of TraM is also composed predominantly of hydrophobic amino acids but contains one possible surface exposed loop. TraM-alkaline phosphatase and TraM-beta-galactosidase fusion proteins supported the hypothesis that TraM has a small cytoplasmic loop. We were unable to detect heterologous complementation between any tra mutation and its homolog from the virB operon of Agrobacterium tumefaciens. PMID- 9200222 TI - Characterization of the cryptic plasmids of the Pseudomonas alcaligenes type strain. AB - The species type strain of Pseudomonas alcaligenes contains three small cryptic plasmids (designated pECB1, 2, and 3) of 7740, 4480, and 2700 bp, respectively. Partial restriction enzyme maps have been constructed for pECB1 and 2 which on this basis do not appear to be related. pECB3 proved refractile to cutting with commonly used restriction enzymes, though it was completely rendered by those enzymes which recognize 4-bp sequences containing only G + C. This suggested that pECB3 is especially rich in these bases. Hybridization studies using labeled pECB2 as probe revealed homology with pECB3 and with regions of the bacterial chromosome, but not with pECB1. A 1214-bp region of pECB2 showed great sequence similarity to the basic replicon of pPS10, a 10-kb Pseudomonas-specific plasmid isolated from Pseudomonas syringae pv. savastonoi. The putative replicon (including the gene for a replication protein) was subcloned and both DNA strands were sequenced. Introduction of the putative replicon into the Escherichia coli plasmid pUC19 created a recombinant vector able to replicate in both E. coli and Pseudomonas spp. Minicell analyses did not reveal any peptides which could be attributed to the remaining region of pECB2 or to pECB1--a finding supported by sequencing studies. Attempts at plasmid curing were unsuccessful. A phenotypic comparison with a non-plasmid-harboring strain of P. alcaligenes, based on nutritional versatility and antibiotic susceptibility, revealed a single difference of note: the type strain alone was able to utilize benzoate for growth. Transformation of the non-harboring strain with pECB1-3, followed by selection on a minimal medium containing benzoate, gave no colonies. The advantages gained by possession of pECB1-3, if any, are at present unknown. PMID- 9200223 TI - Isolation and characterization of a plasmid from Lactobacillus fermentum conferring erythromycin resistance. AB - Lactobacillus fermentum is a lactic acid bacterial species commonly found in the digestive tracts of pigs and rodents and also present in man. We characterized a 5.7-kb plasmid, pLEM3, conferring erythromycin resistance, which was isolated from a porcine strain of L. fermentum. Plasmid pLEM3 established efficiently in L. fermentum, conferred high-level erythromycin resistance (MIC > 1 mg/ml), and was segregationally stable. A deletion derivative of pLEM3, called pLEM5, was constructed and found to be as genetically stable as the parent. A multiple cloning site was inserted into pLEM5, generating plasmid pLEM7. Nucleotide sequence determination of pLEM5 revealed similarities with known genes. The replicon itself is a member of the pC194 family of rolling circle plasmids. The region responsible for erythromycin resistance was 98.2% identical to the erm gene of conjugative transposon Tn1545. PMID- 9200224 TI - Identification of two sequence elements associated with the gene encoding the 24 kDa crystalline component in Bacillus thuringiensis ssp. fukuokaensis: an example of transposable element archaeology. AB - A 6.5-kb fragment of plasmid DNA from Bacillus thuringiensis (Bt) ssp. fukuokaensis that encodes a 24-kDa crystalline component was analyzed to identify additional open reading frames (orfs). A novel Bt IS240-like element was found upstream of this gene and is considered to be a vestige of a once active insertion sequence due to a stop codon that interrupts the long orf encoding the putative transposase. This element was bounded by 17-bp terminal inverted repeats that defined the length of the insertion sequence as 802 bp. Further upstream of this element two tandem overlapping and out of phase open reading frames (orfX and orfY) were identified which represent the first example of an IS150-like element in Bt containing both orfs. orfX and orfY are not bounded by terminal inverted repeats but are associated with a gene encoding a putative site-specific recombinase of a type found in Staphylococcus aureus Class II transposons but not previously in Bt. PMID- 9200225 TI - Comparative evaluation of three commercial broiler stocks in hot versus temperate climates. AB - Hot climate is a major limiting factor of broiler production in tropical and subtropical regions. The use of standard stocks in hot climates may result in large economic losses because genotypes selected in temperate climates may respond differently to the high ambient temperatures in hot regions or seasons. The summer and fall in Izmir, Turkey, provided the natural hot and temperate climates, respectively, for this study. Broiler chicks were obtained from three commercial stocks, all bred in temperate climates. Male and female chicks, 60 per pen, were housed in four pens per stock per season. Individual BW was determined at hatch, and at 4 and 7 wk of age. Feed consumption and efficiency were determined per pen. Feathering was scored at 4, 5, and 6 wk of age. Body temperature was measured twice on three birds per sex per pen, 16 h and immediately before slaughter, and feather weight was determined for each of these birds. The two seasons clearly differed in ambient temperature at the broiler house, and consequently, BW at 7 wk was significantly lower in the summer than in the fall in all stocks, with an average reduction of 23%. The season effect was largest (33.5%) on BW gain from 4 to 7 wk, along with 23 and 15% reductions in feed consumption and efficiency, respectively, during these 3 wk. A significant season by stock interaction was detected for BW gain from 0 to 4 wk and 4 to 7 wk. The three stocks exhibited similar 4- to 7-wk BW gains under the temperate fall climatic conditions, but differed significantly in the summer. These differences were not related to normal differences in feather coverage or body temperature, suggesting that standard broiler stocks must be tested in hot climates in order to find the one most suited to these conditions. PMID- 9200226 TI - Performance of naked neck and normal broilers in hot, warm, and temperate climates. AB - Chickens suffer at high ambient temperatures because their feather coverage hinders internal heat dissipation. Naked neck broilers (Na/na) and their normally feathered sibs (na/na) were evaluated in three natural climates. Three experiments were conducted in Turkey, during the summer in the extremely hot region of Adana (Ad-Sm), and in the moderate region of Izmir during the spring (Iz-Sp) and summer (Iz-Sm), always following the same experimental protocol. Ambient temperatures averaged 19, 28, and 32 C in Iz-Sp, Iz-Sm, and Ad-Sm, respectively. About 300 birds per genotype were included in each experiment. Feather weight was lower by about 20% in Na/na broilers than in na/na ones, independent of climate, sex, and age (6 or 7 wk). The Na/na broilers exhibited higher breast weight in all cases, from 2.5 to 10.9% higher than their na/na counterparts. Body weight gain from 4 to 7 wk (BWG4-7) clearly reflected the differences in ambient temperature among climates. The effect of the Na/na genotype on BWG4-7 interacted with climate and sex. In the hottest climate (Ad Sm), both male and female Na/na broilers exhibited a highly significant advantage over their na/na counterparts. In the more moderate climate (Iz-Sm), the Na/na genotype exhibited superior growth only among males, and the magnitude of this advantage was lower than in Ad-Sm. In the cool temperate climate (Iz-Sp), BWG4-7 and BW7 (BW at 7 wk) means were similar for both genotypes. In Iz-Sp, feed efficiency (FE) of the Na/na birds was lower by about 4%, but in the two summer climates (Iz-Sm and Ad-Sm), FE of the Na/na birds was about 9% higher than that of their na/na counterparts. Body temperature was lower in the Na/na broilers than in their na/na counterparts; in all cases, the difference increasing with ambient temperature. The results indicate that the reduction in feather coverage provided relative heat tolerance, and therefore, under hot climates the Na/na broiler were superior to their normally feathered counterparts. It is concluded that naked neck broilers should be preferred in hot climates. PMID- 9200227 TI - Educational opportunities and challenges in poultry science: impact of resource allocation and industry needs. AB - Because the number of Poultry Science departments in the U.S. has declined dramatically, and because scientist years and research funding for poultry, relative to other commodities, have also declined, a survey of poultry meat companies was conducted. Objectives of the survey were: to evaluate corporate concern over the status of Poultry Science departments, to categorize hiring patterns, to determine expectations for prerequisite skills of graduates, and to ascertain attitudes toward hiring of Associate-degreed students (A.S.). A two page survey was distributed to corporate Vice Presidents or Directors of Human Resources of the 17 largest broiler and 10 largest turkey companies. When asked to gauge the difficulty they encountered in locating adequate numbers of Poultry Science graduates, 83% noted at least some difficulty. All respondents indicated concern over the loss of poultry programs in the U.S. and 44% noted "extreme" concern. There appears to be little resistance to hiring 2-yr A.S. degree graduates in Poultry Science. The relative scarcity of these programs is demonstrated by the fact that only one-third of the respondents had ever hired A.S. degree graduates. However, greater than 80% of the firms indicated they would hire these students. Finally, communication and business skills were more highly rated by human resources management than technical ability in Poultry Science. Given these results, academic programs must: develop curricula that reflect market-place expectations, enhance the efficiency of resource utilization, embrace new technologies that provide novel methods for information delivery, and reassess cooperative linkages among industrial and governmental organizations. PMID- 9200228 TI - Water consumption by broilers in high cyclic temperatures: bell versus nipple waterers. AB - Broilers were maintained on litter in environmental chambers to study water consumption from bell and nipple waterers. The chambers were set at high cyclic temperatures of either 24-35-24 C or 24-32-24 C daily cycles in three trials. Water consumption was recorded by computer each 30 min and calculated as a percentage of body weight. Daily water consumption from nipples was always less than from bell waterers. Water consumption by quarter-day revealed that consumption from nipples was often similar to that from bell waterers during the lowest temperatures but was less during the periods of highest temperatures. Further study revealed that water consumption from nipple waterers was related to the height of the nipples such that consumption was greater for lower nipples. The results suggest that panting broilers have difficulty drinking from high nipple waterers. PMID- 9200229 TI - Estimation of the composition of broiler carcasses from their specific gravity. AB - An experiment was conducted to quantify the relationships between broiler carcass specific gravity and chemical composition (percentage moisture, percentage lipid, percentage protein). Carcasses of widely varying compositions were produced by feeding several dietary protein and energy combinations (52 to 64% moisture, 0.6 to 2.5% ash, 1.6 to 11.7% lipid, and 4.9 to 8.0% nitrogen). Very strong relationships were found between percentage moisture and percentage lipid (r = 0.969) and percentage moisture and percentage N (r = 0.968). Strong relationships were found between specific gravity and percentage lipid (r = -0.872) and specific gravity and percentage N (r = 0.857). Specific gravity is recommended as a means to estimate carcass fat in broiler chickens. PMID- 9200230 TI - Atypical Escherichia coli strains and their association with poult enteritis and mortality syndrome. AB - To date, no definitive etiology has been described for Poult Enteritis and Mortality Syndrome (PEMS). However, two atypical Escherichia coli colony types are isolated consistently from moribund and dead poults afflicted with PEMS. To test the infectivity of these E. coli strains, poults were placed into floor pens in three isolation treatment rooms: 1) CONTROL: no bacterial challenge, 2) E. coli colony Types 1 or 2 posthatch oral challenge: 10(8) cfu/per poult at 1 d, and 3) E. coli colony Types 1 or 2 posthatch oral challenge: 10(8) cfu/per poult at 6 d. Daily intramuscular injections of cyclophosphamide (100 micrograms per poult) from 1 to 5 d posthatch were given to half of the poults in each treatment. Atypical E. coli challenge caused BW depression, and cyclophosphamide treatment exacerbated the response. All E. coli-challenged poults developed diarrhea similar to PEMS. Mortality was increased by both atypical E. coli colony types, but at 21 d E. coli colony Type 2 caused greater mortality than colony Type 1. With cyclophosphamide treatment, mortality was exacerbated with both colony types, but colony Type 2 at 1 d caused the greatest mortality. Ultrastructural damage to ileum epithelium cell microvilli and subcellular organelles indicated that part of the BW depression could be attributed to malabsorption of nutrients. It was concluded that the atypical E. coli colony Types 1 and 2 play a significant role in the PEMS disease. PMID- 9200231 TI - Effect of Santoquin and oxidized fat on liver and intestinal glutathione in broilers. AB - Experiments were conducted to determine effects of Santoquin (ethoxyquin) and oxidized fat on liver and intestinal reduced (GSH) and oxidized (GSSG) glutathione, and pulmonary hypertension syndrome (PHS) mortality. Male broilers were randomly assigned in a 2 x 2 factorial consisting of 3.5% normal (NF) or oxidized (OxF) fat with or without ethoxyquin (E). Body weights and feed intake were monitored weekly, and tissues obtained at 3 and 7 wk for GSH and GSSG analysis. Compared to the NF group, NF/E gained more weight during the starter (0 to 3 wk), but not the grower (4 to 7 wk) period. Birds fed NF/E or NF exhibited greater feed efficiency in the starter period and greater gains during the starter and grower periods than birds fed OxF or OxF/E. No differences in PHS mortality between treatments were observed. Birds fed OxF exhibited lower liver GSSG at 3 wk than the other groups, but there were no differences in liver GSH. Duodenal GSH was higher in birds fed OxF/E than in birds of NF group at 3 and 7 wk. Ileal GSH was higher at 3 wk in OxF/E birds than in OxF birds, but no differences were observed at 7 wk. All tissues exhibited higher GSH levels at 7 wk than at 3 wk. Birds fed ethoxyquin, regardless of fat source, exhibited higher duodenal GSH at 3 and 7 wk and higher ileal GSH at 3 wk than birds that did not receive ethoxyquin. Higher GSH would be beneficial by enhancing protection of intestinal cells to deleterious effects of toxins or other forms of oxidative stress. PMID- 9200232 TI - Role of contact and genetic transmission of endogenous virus-21 in the susceptibility of chickens to avian leukosis virus infection and tumors. AB - The role of contact and genetic transmission of endogenous virus-21 (EV21) on response of chickens to avian leukosis virus (ALV) infection and tumors was studied. F1 progeny of a cross between RPRL late-feathering (LF) line EV21+ males and RPRL early feathering (EF) line 15B1 females harboring or lacking EV21 were used. The EF chicks lacking EV21 were inoculated with a field strain of subgroup A ALV at hatch and contact exposed to LF, EV21+ hatchmates for various time intervals. In a second experiment, EV21 contact-exposed and unexposed EF chicks as well as LF, EV21+ hatchmates were inoculated with ALV at various ages. Chickens were tested for ALV-induced viremia and antibody and were observed for tumors until 24 wk of age. Antibody to EV21 in EF chickens contact-exposed to LF, EV21+ hatchmates varied from 10 to 65%, and was detected by 10 wk of age. By 24 wk of age, ALV-induced viremia and tumors in EF chickens varied from 5 to 30%, and from 15 to 32%, respectively, regardless of exposure to EV21. The incidence of ALV-induced tumors was significantly higher in LF chickens genetically infected with EV21 than in EV21 contact-exposed or unexposed EF chickens, but only in chickens inoculated with ALV at hatch. The data suggest that contact infection with EV21 has no influence on ALV infection and tumors. The data also suggest that genetic transmission of EV21 may increase susceptibility of chickens to ALV infection and tumors following infection with ALV at hatch, but not at 4 wk of age or older. PMID- 9200233 TI - Immunogenic characterization of a tissue culture-derived vaccine that affords partial protection against avian coccidiosis. AB - The immunogenicity of a tissue culture-derived vaccine generated from an Eimeria tenella-infected cell line in a serologically defined bird line, and the ability to confer protection against homologous challenge in young chicks was examined. The cell line, SB-CEV-1/F7, was infected with E. tenella sporozoites and the resulting 72-h postinfection cell-free supernatants were adjuvanted and used to immunize Leghorn chicks homozygous for the B19 haplotype. Peripheral blood and splenic lymphocytes from these immunized birds proliferated in vitro in response to both sporozoite and SB-CEV-1/F7 tissue culture-derived parasite antigens. In addition, splenic immune lymphocytes obtained from birds previously exposed to E. tenella in vivo responded to these tissue culture-derived parasite antigens in vitro. To evaluate the efficacy of the vaccine, B19B19 chicks were vaccinated s.c. with adjuvanted 72-h postinfection cell-free supernatants or an ammonium sulfate precipitate derivative thereof, orally boosted, and then subjected to homologous parasite challenge at 10 d of age. The level of protection (body weight gain, cecal lesions) was assessed 6 d after challenge. Performance results from four battery trials demonstrated that vaccinated birds were significantly protected against weight loss compared to unimmunized, challenged controls. In addition, in two of the four trials, vaccinated birds were significantly protected against lesions. These results provide strong evidence that tissue culture-derived parasite antigens obtained from the E. tenella-infected SB-CEV 1/F7 cell line are immunogenic in birds and can provide partial protection against E. tenella clinical coccidiosis. PMID- 9200234 TI - Enhancement of humoral and cellular immunity by vitamin E after embryonic exposure. AB - In the present study, the amnion of turkey and chicken embryos were injected 3 d prior to hatch with different levels of vitamin E (VE). In Experiments 1 and 2, turkey embryos received 10, 20, and 30 IU of VE. In Experiment 3, broiler embryos received 10 IU VE. In all three experiments, sham-injected control embryos (0 IU VE) received 300 microL of saline. In Experiments 1 and 2 (turkey embryos), 20 and 30 IU of VE reduced (P < or = 0.05) percentage hatchability below that of controls. At hatch, poults exhibited a dose related increase (P < 0.05) in plasma VE levels. Mean BW gain up to 35 d and relative bursa of Fabricius and spleen weights were not different among treatment groups. When challenged at 7 d posthatch, total (P < 0.05) and IgM (P < 0.08) anti-SRBC antibodies were higher in 10 IU VE poults than in controls. Immunoglobulin G levels did not differ among the treatment groups. Poults in the 10 IU VE group had higher (P < 0.002) numbers of Sephadex-elicited inflammatory exudate cells, as well as a greater percentage of phagocytic macrophages (P < 0.0001). Additionally, the numbers of SRBC per phagocytic macrophage were greater (P < 0.001), than in control poults at 4 wk of age. In Experiment 3, chick embryos exposed to 10 IU VE, exhibited no differences in hatchability, BW gain, or bursal and splenic weights from the sham-exposed group. However, total and IgM antibody responses against SRBC were greater (P < 0.01) in the 10 IU VE group at 7 d postinjection. A secondary SRBC challenge given at 14 d after primary injection resulted in higher total (P < 0.07) and IgG (P < 0.04) antibody responses in the 10 IU VE chicks than in the controls. Similarly, broiler chicks (10 IU VE) had more Sephadex-elicited abdominal exudate cells (P < 0.07), and greater macrophage phagocytic potential (P < 0.0001). In ovo VE exposure (10 IU) also increased nitrite production (P < 0.04) by chick macrophages. The results from this study demonstrated an enhanced antibody and macrophage response and suggest that in ovo exposure with VE may improve posthatch poult and broiler quality. PMID- 9200235 TI - Nutrition of the broiler chicken around the period of compensatory growth. AB - Three experiments were conducted with cage-reared broilers to 21 d following nutrient restriction from 6 to 12 d age. In Experiment 1, birds were full-fed from 6 to 11 or given 50% of ad libitum intake on a daily basis, or 100% of ad libitum intake on a daily basis when the diet was diluted 50% with oat hulls. Birds were not able to fully recover body weight depression by 21 d, although birds previously restricted, by whatever method, were more efficient (P < 0.01) in overall energy intake:body weight gain. Prior feed restriction had no effect on ability to metabolize diet energy (P > 0.05), although these birds did exhibit increased nitrogen retention compared to birds full-fed from 6 to 11 d. In a second experiment, birds were fed diets with 1.25, 1.38, 1.51, 1.63, 1.76, or 1.88% lysine in the realimentation diet from 12 to 21 d. Lysine level had no effect on growth rate or feed efficiency (P > 0.05) for full-fed birds; however there was a linear (P < 0.05) decline in growth rate from 12 to 21 d in response to extra dietary lysine for the birds previously feed restricted from 6 to 12 d. In a third experiment, birds were fed diets varying in energy (3,000 to 3,300 kcal/kg) or protein (22 to 29% CP) from 12 to 21 d following ad libitum vs 50% feed restriction from 6 to 11 d age. Protein level of the diet had little effect on performance traits to 21 d, although there was an indication of improved growth in response to the higher energy concentration. Birds full-fed from 6 to 11 d showed increased liver size at 21 d when fed more protein, although the converse was true for the restricted birds (P < 0.05). The growth response to diet energy was associated with increased carcass fatness. In general, there does not seem to be any advantage to manipulating diet formulation during realimentation of birds previously nutrient-restricted. PMID- 9200236 TI - The response of broiler chickens and turkey poults to dietary energy supplied either by fat or carbohydrates. AB - The efficacy of fat and carbohydrates as energy sources was compared in 1- to 4- and 4- to 7-wk-old broiler chickens and in 16- to 19-wk-old turkeys. An increase in dietary energy by carbohydrate was made by a graded replacement of wheat bran by wheat. Energy was increased by fat through a graded replacement of soybean hulls with refined soybean oil. In the experiments with broiler chickens, the feed efficiency responses to added energy were observed within the entire range of dietary energy tested, with no significant differences between the responses to carbohydrate and fat as energy supplements. The growth response to energy from either source appeared to be characterized by diminishing returns in the chicken. In the 16- to 19-wk-old turkeys, the growth and feed efficiency responses were linear within the range from 2,650 to 3,250 kcal/kg. In chickens and in turkeys, the growth and feed efficiency responses to energy supplied by fat were indistinguishable from those of carbohydrates. In chickens, the fractions of abdominal fat and pectoral muscle were not affected significantly by the energy density and source. PMID- 9200237 TI - The response of broiler chickens and turkey poults to steam-pelleted diets supplemented with fat or carbohydrates. AB - The responses of growth and feed efficiency to pelleted feed was investigated in 4- to 7-wk-old broiler chickens, and in 8- to 12- and 16- to 20-wk-old turkeys. In all cases, the growth and feed efficiency responses were linear within the ranges of dietary energy tested. When energy was added by carbohydrate supplementation, weight gain and feed efficiency responses were parallel for both mash and pellets, but due to the growth response to pellets, the elevation was higher for pellets than for mash. When energy was added by fat, the growth response to pellets also resulted in an increase in function elevation but the slope of the response was lower than in mash feeding, possibly due to a decline in pellet quality as dietary fat increased. Grinding of pellets completely abolished the growth and feed efficiency responses observed when the physical form was preserved. In chickens, comparisons of ground pellets to mash suggested some decline in nutritional quality due to the process of pelleting when either carbohydrates or fat were increased in the diets. In both chickens and turkeys, the feeding of pelleted diets resulted in an increase in abdominal fat. PMID- 9200239 TI - Diphasic allometric growth of body components in white Leghorn pullets fed ad libitum and restricted diets. AB - Growth patterns of crude fat (cfat; ether extract), CP, ash, and water in White Leghorn pullets that ate ad libitum and restricted (feed or lysine) diets were analyzed by mono- and diphasic allometric functions, with plucked empty body mass (EBM) or fat-free EBM (FFEBM) as the independent variates. In general, the diphasic model provided a better fit to the data than the monophasic model, indicating that the allometric slope (beta) changes at a certain level of development. Data demonstrate that, under an ad libitum feeding regimen, cfat growth is proportionally (beta = 1) related to growth of the fat-free body, up to about 500 g FFEBM (pullets aged 8.5 wk). From 500 g FFEBM onwards, the beta changed to about 2, illustrating the late maturation of cfat compared to FFEBM. For feed-restricted groups, slopes and transitions for cfat growth varied from those of the control group, according to changes in dietary supply. Diphasic relationships between each fat-free body component and EBM in each treatment had different slopes for first and second phases (about 1 and 0.70). Differences, however, were mainly due to the diphasic growth pattern of cfat per se, and diminished largely if FFEBM instead of EBM was chosen as the independent variable (about 1 and 0.95). Incorrect statements on body component growth in restricted birds, due to the "masking" role of varying fat deposition rates, can be avoided if relative growth was expressed as a function of the fat-free body instead of BW or EBM. Diphasic growth relationships among the fat-free body components were not affected by feeding regimen, but by age. The beta 1 were about unity for the relationship between each fat-free component and FFEBM, suggesting a constant composition of the fat-free body in the first phase. In the second phase, CP accrued relatively faster than ash and water (beta 2 of 1.10 vs 0.94 and 0.91). Results indicate that the changes in growth pattern of fatty and fat-free tissue, both relative to the fat-free body, had not been revealed if a monophasic allometric model instead of a diphasic had been used. PMID- 9200238 TI - Interactive effects of betaine and monensin in uninfected and Eimeria acervulina infected chicks. AB - Three experiments (Exp.) were conducted to evaluate the interactive effects of dietary betaine (BET) and monensin (MON) in uninfected or Eimeria acervulina infected chicks. The treatments were replicated with six (Exp. 1) or five (Exp. 2 and 3) pens of five chicks each. The experimental periods lasted 9 (Exp. 1 and 2) or 10 (Exp. 3) d each and the coccidiosis infections were established on Day 2 (Exp. 1 and 2) or Day 3 (Exp. 3) of the experiment. Average initial weight of the chicks was 101, 73, and 68 g in Exp. 1 to 3, respectively, and the initial age of the chicks was 5 (Exp. 1) or 4 (Exp. 2 and 3) d. A corn-soybean meal basal diet was used in each experiment. In Exp. 1, the effect of dietary BET (0, 0.1, or 0.5%) in uninfected or coccidiosis-infected (COC; 5 x 10(5) sporulated E. acervulina oocysts) chicks was investigated. In Exp. 2, the interactive effects of BET (0 or 0.1%) and MON (0 or 55 ppm) in uninfected or COC chicks were investigated in a 2 x 2 x 2 factorial arrangement of treatments. Experiment 3 was identical to Exp. 2, except the level of MON was 110 rather than 55 ppm. In Exp. 1, 2, and 3, COC reduced (P < 0.01) gain, feed intake (FI), feed efficiency (GF), and plasma carotenoid concentration (CAR) and increased (P < 0.01) lesion score (LS). In Exp. 1, gain and FI were decreased in uninfected chicks fed 0.1% BET but gain and FI were increased in COC chicks fed 0.1% BET (COC x BET quadratic, P < 0.01). Dietary BET linearly increased (P < 0.05) GF. In Exp. 2 and 3, MON increased (P < 0.01) gain, FI, GF, and CAR and decreased (P < 0.01) LS of COC chicks, but MON had no effect in uninfected chicks (COC x MON, P < 0.01). In Exp. 2, GF was increased more in chicks fed both MON and BET than in chicks fed MON (BET x MON, P < 0.06). In Exp. 3, BET increased GF of uninfected chicks fed MON and of COC chicks not fed MON (COC x BET x MON, P < 0.02). Betaine may have an effect on E. acervulina-infected chicks, but there is no conclusive evidence to indicate that the efficacy of MON is improved when fed in combination with BET. PMID- 9200240 TI - Effect of strain and age of the broiler breeder female on incubation time and chick weight. AB - Two experiments were conducted to evaluate effects of strain [five in Experiment (Exp.) 1 and six in Exp. 2)] and age (29, 47, and 57 wk in Exp. 1 and 29, 41, and 52 wk in Exp. 2) of commercial broiler breeders on incubation time and chick weight. Highly significant differences in egg weight were found among strains in both Exp. After adjusting for effects of egg weight, significant effects of strain, age, and their interactions were found on incubation time, egg weight at transfer, and chick weight at hatch in Exp. 1, but not in Exp. 2. Mean incubation times varied among strains from 496.6 to 498.8 h in Exp. 1 and from 499.3 to 501.9 h in the second experiment. In Exp. 1, incubation time decreased from 498.6 h when breeders were 29 wk to 494.8 at 47 wk, whereas in Exp. 2, it decreased from 510.5 h at 29 wk to 495.1 h at 41 wk. This decrease also resulted in a negative correlation between egg weight and incubation time. Differences due to strain and age were found for yolk and albumen percentage and yolk: albumen ratio. Percentage yolk was 27.2 and 32.7% and percentage albumen was 60.1 and 55.9% in eggs from 29 to 52 wk breeders, respectively. Shell percentage was significantly affected by strain. Strain by age interactions were found for each response in Exp. 1 but only for set and chick weight in Exp. 2. Differences among incubators were found only for incubation time; interactions of incubation time and strain and age were also detected. Results indicate that genotype, age of the female breeder, and incubator should be considered along with their interactions to obtain optimum hatching performance. PMID- 9200241 TI - Relationship between aerobic bacteria, salmonellae and Campylobacter on broiler carcasses. AB - Broiler carcasses were removed from commercial processing lines immediately after defeathering, before chilling, and after chilling to determine whether any relationship exists between aerobic bacteria and the human enteropathogens salmonellae and Campylobacter. In two experiments, a whole carcass rinse procedure was used to sample 30 carcasses after defeathering, 90 carcasses before chilling, and 90 carcasses after chilling, for a total of 210 different carcasses. Aerobic bacteria and Campylobacter spp. were enumerated and the incidence of salmonellae was determined. Salmonellae and Campylobacter incidences were 20 and 94%, respectively, for all carcasses sampled. After picking, neither salmonellae-positive nor Campylobacter-positive carcasses had mean aerobic most probable number (MPN) values that were different from carcasses negative for those organisms. Immediately before chilling, aerobic and Campylobacter counts were 7.12 and 5.33 log10 cfu per carcass, respectively. Immersion chilling reduced aerobic counts by approximately 1.8 log and Campylobacter by 1.5 log, with no change in salmonellae-positive carcasses. There was no difference in aerobic or Campylobacter counts between carcasses that were positive or negative for salmonellae at any of the sampling locations, nor was any correlation found between levels of aerobic organisms and Campylobacter. Carcasses with aerobic counts above the mean or more than one standard deviation above the mean also failed to show any correlation. Discriminant analysis indicated error rates as high as 50% when numbers of aerobic bacteria were used to predict incidence of salmonellae or Campylobacter on individual carcasses. Aerobic bacteria are not suitable as index organisms for salmonellae or Campylobacter on broiler carcasses. PMID- 9200242 TI - The relationship of broiler breast meat color and pH to shelf-life and odor development. AB - Experiments were conducted to compare the shelf-life of dark-colored and light colored broiler breast meat. In each of three trials, 100 breast fillets were obtained from a commercial processing plant and subjectively categorized as "dark" or "light". The 100 fillets were then objectively evaluated for C.I.E. color values (lightness, redness, and yellowness). The fillets were separated into five storage groups, with each group containing 10 dark and 10 light fillets, and the fillets were held at 3 C for 0, 3, 6, 9, and 12 d. On each sampling day, fillets were evaluated in duplicate for psychrotrophic plate count (PPC), capacitance detection time (CDT), pH, and subjective odor evaluation. Dark fillets had significantly (P < 0.05) lower lightness values (L*), higher redness values (a*), lower yellowness values (b*), and higher pH values. Regression coefficients for odor scores resulted in darker fillets having significantly (P < 0.05) higher slopes than lighter-colored fillets even though intercept values were similar. Significant correlations existed between pH and color as well as odor, CDT, and PPC. These data suggest that darker broiler breast meat fillets have a shorter shelf-life than lighter breast fillets; the shorter shelf-life may be due to differences in pH. PMID- 9200243 TI - Muscle metabolism and meat quality of Pectoralis from turkeys treated with postmortem electrical stimulation. AB - This experiment was conducted to evaluate the effects of electrical stimulation (ES) on muscle metabolism and breast meat quality in turkeys. Thirty-six turkey hens were either ES at the neck in a saline bath (570 V, 450 mA, AC, 60 Hz, 2 s on 1 s off for 10 pulses) or used as unstimulated controls. One breast fillet from all carcasses was harvested at 2 h postmortem. The opposite fillet was harvested from the ES carcasses at 8 h postmortem and from the unstimulated controls at either 8 or 24 h postmortem. All fillets were sampled at time of deboning for expressible moisture, pH, R-value, gravimetric fragmentation index (GFI), and sarcomere length. The remainder of the fillet and the samples for GFI and sarcomere length were aged on ice until 24 h postmortem. After aging, fillets were analyzed for cook loss and shear value. Color was measured at time of deboning and at 24 h postmortem. Electrical stimulation accelerated rigor mortis development as indicated by significantly lower pH values and higher R-values at 2 h postmortem when compared to control fillets. The pH and R-values of the 2-h ES treatment were not significantly different from the 8-h ES, 8-h controls, or the 24-h controls. Fillets from carcasses that were ES and deboned at 2 h had significantly longer sarcomeres than the 2-h controls; however, there were no significant differences between the 2-h ES and the 8-h ES treatments, 8-h controls, or the 24-h controls. Although ES accelerated muscle metabolism at 2 h postmortem, it had no effect on shear value, expressible moisture, cook loss, GFI, L*, or a* color values. These results suggest that this postmortem ES system would not benefit turkey processors. PMID- 9200244 TI - Effects of stunning duration on quality characteristics of early deboned chicken fillets. AB - The objective of this study was to determine effects of electrical stunning duration on quality of broiler chicken fillets. Seventy-two broiler chickens were electrically stunned for 0, 2, 4, 6, 8, or 10 s, slaughtered, and chilled. After 1 h post-mortem, both pectoral muscles were excised and cooked. Cooking loss, pH, cooked color values, and shear values were measured. As stunning time increased, pH and shear values significantly increased. Except for a small but significant increase in yellowness, color values were unaffected by stunning duration. Cooking loss was unaffected by stunning duration. These data indicate that stunning duration can affect post-mortem muscle metabolism as measured by pH change. Therefore, control of the process of slaughtering broilers requires careful regulation of stunning duration. PMID- 9200245 TI - Relative contributions to the net joint moment for a planar multijoint throwing skill: early and late in practice. AB - The purpose of this investigation was to determine, for a planar, multijoint throwing skill, if the relative contributions of the components of the net joint moment (NJM) at the elbow and shoulder change after practice. Each participant (N = 7) performed 200 throwing trials equally distributed across 5 consecutive days. Each participant threw a 0.15-kg ball as far as possible using the nondominant arm while the motion of the throwing arm was restrained to a horizontal plans. From video data and body segment inertial estimations, NJMs and NJM components (i.e., generalized muscle moments and motion-dependent moments) were calculated for selected early and late practice trials. Performance (throwing distance) showed an expected improvement from early to late practice. The dynamics analysis indicated that participants increased average NJMs and NJM components at both joints. However, the relative contribution of NJM components, expressed as ratios of those components to the NJM at each joint, did not change after extended practice. Restraining the throwing arm to a horizontal plane may partly explain why no changes were found in the relative contributions of NJM components. The lack of change in moment ratios support a motor strategy of scaling joint moments for faster movements. PMID- 9200246 TI - A multilevel analysis of school factors associated with health-related fitness. AB - School factors associated with health-related fitness, using the National Children and Youth Fitness Survey II (NCYFS II) data, have been examined by Pate and Ross (1987), but the hierarchical structure of the data was ignored in the data analyses. The purpose of this study was to reanalyze the NCYFS II data using the hierarchical linear model and reexamine the effects of school physical education (PE) and other factors on children's 1-mile run/walk performance. Only two of five "significant" school factors were confirmed by this study, suggesting that heightened probabilities of Type I errors might have occurred in the previous study. Two confirmed key characteristics of school PE programs that led to improvement of children's cardiovascular endurance were PE specialists and the administration of fitness tests. For a hierarchical data structure, both experimental and observational units should be considered in the data analysis. The hierarchical linear model, which not only provides more accurate individual prediction but also takes group effect into account, proved to be an appropriate analytical model in analyzing the hierarchical data. PMID- 9200247 TI - Recalling demonstrated and guided movements using imaginary and verbal rehearsal strategies. AB - This study investigated the recall of movement patterns presented either by demonstration or guided movement with vision eliminated. Participants were instructed to rehearse and remember each of the 12 patterns using one of four strategies: imagery, verbal labeling, imagery and verbal labeling, or no rehearsal strategy (i.e., control condition). Recall was better for patterns that were demonstrated than for those presented via guided movement. In addition, more patterns were remembered if a combination of imagery and verbal labeling were employed as a rehearsal strategy compared to using imagery alone. These results are discussed using Annett's (1994) model showing the relationships between action, language, and imagination in the acquisition of motor skills. PMID- 9200248 TI - The effect of knowledge of results delay and the subjective estimation of movement form on the acquisition and retention of a motor skill. AB - This study examined the effects of knowledge of results (KR) delay and subjective estimation of movement form on the acquisition and retention of a motor skill. During acquisition, four groups of participants performed 60 trials of a throwing accuracy task under the following conditions: (a) immediate KR, (b) delayed KR, (c) immediate KR + form estimation, and (d) delayed KR + form estimation. Retention tests of throwing accuracy and outcome error estimation in the absence of visual KR were administered 5 min and 24 hours following acquisition. Throwing accuracy was significantly higher during acquisition but significantly lower during retention for immediate-KR participants than for delayed-KR participants. However, participants who estimated their movement form during acquisition produced significantly higher throwing accuracy and lower estimation error during retention than those who did not. PMID- 9200249 TI - Development of an instrument to assess cognitive processes in physical education classes. AB - Using the mediating process paradigm (Doyle, 1977) as a framework, the need to develop reliable and valid instruments for assessing cognitive processes is apparent, and that was the purpose of this study. Participants (N = 819) completed the Cognitive Processes Questionnaire in Physical Education (CPQPE), as well as three other instruments addressing dispositional goal orientation, perception of motivational climate, and beliefs about causes of success in physical education. The five-factor structure that emerged in an exploratory factor analysis produced an acceptable fit with the data in the confirmatory factor analysis. The subscales of the CPQPE were related to a task-involved goal perspective and the belief that success is attributed to motivation and effort. The results indicate the CPQPE is a valid and reliable instrument that can provide valuable information about the teaching and learning process. PMID- 9200250 TI - The physiological responses to walking with and without Power Poles on treadmill exercise. AB - Power Poles are specially constructed, rubber-tipped ski poles designed for use during walking. Using Power Poles simulates the arm motion of cross-country skiing, thus increasing the muscle mass used during walking. This study investigated the potential increases in exercise intensity and energy cost associated with the use of walking poles. Thirty-two apparently healthy volunteers (16 men and 16 women) between the ages of 19 and 33 years participated. Each completed a treadmill maximal oxygen consumption (VO2max) test and two randomly assigned, submaximal walking trials (one with poles and one without poles) on separate days. Each walking trial was conducted on a level treadmill, for 20 minutes, at an identical self-selected pace. Expired gases, heart rate in beats per minute (bpm), and ratings of perceived exertion (RPE) were recorded each minute. Results between trials were compared using repeated measures analysis of variance and Tukey's post hoc tests. It was found that walking with poles resulted in an average of 23% (4.4 ml.kg-1.min-1) higher VO2, 22% higher caloric expenditure (1.5 kcal.min-1), and 16% (18 bpm) higher heart rate responses compared to walking without poles on a treadmill. RPE values averaged 1.5 units higher with the use of the poles, and the pattern of responses between conditions was similar for men and women. It is concluded that the use of Power Poles can increase the intensity of walking at a given speed and, thus, may provide additional training benefits to walkers. PMID- 9200251 TI - Summary knowledge of results and task processing load. PMID- 9200252 TI - How close is too close for precise knowledge of results? PMID- 9200253 TI - Running economy: comparison of body mass adjustment methods. PMID- 9200254 TI - Indication, incidence and management of blood transfusion during sinus surgery: a review over 12 years. AB - The number of reports about blood transfusion-related HIV and hepatitis virus infections is increasing, presently. Thus, it should seriously be considered to inform the patient of any anticipated blood loss necessitating a transfusion of blood products. This is especially necessary for surgical procedures with only a low risk for high blood loss, such as endonasal surgery as a common otorhinolaryngological procedure. However, reports about the incidence of blood transfusion during this kind of surgery are very rare. The medical histories of 6,296 patients who underwent sinus surgery between 1982-1993 in the Department of Otorhinolaryngology at the University of Kiel were analyzed. Twenty-nine of these patients received a transfusion. Risk factors for required blood, the necessity of pre-operative information and the recommendation policy for pre-operative donation of autologous blood are discussed. The intra-operative blood losses of 120 patients who did not require a transfusion and who underwent sinus surgery in 1986 and 1989, were analyzed. The transfusion rate was 0.46% on average during the 12-year period. The incidence of blood transfusion and the amount of intra operative blood loss decreased after combination of endonasal surgery with controlled intra-operative hypotension (0.07%; p < 0.01). Risk factors for the necessity of a transfusion in these cases were extensive polyposis and purulent exacerbation of the disease. There are risks for a blood transfusion in endonasal surgery. Every transfusion carries a certain risk for the infection with HIV or hepatitis, therefore every patient should be informed about the possibility of a blood transfusion prior to the operation. Endonasal microscopic sinus surgery performed by well-trained surgeons, combined with controlled intra-operative hypotension lowered the risk for a transfusion significantly (p < 0.01). PMID- 9200255 TI - The nasal septum and the development of the midface. A longitudinal study of a pair of monozygotic twins. AB - The development of the nose and the growth of the midface has been followed in a pair of identical twins. One of them (twin A) had nasal septum destruction after septal haematoma and abscess at the age of 7 years, and was treated by immediate implantation of homologous septal cartilage from a tissue bank. From 7-17 years of age the growth and development of the nose and face were followed. Lateral cephalograms, photographs, acoustic rhinometry and rhinoscopy were performed. Twin B presented a normal nasal and facial growth and served as control. Twin A developed a saddle nose, an upward displacement of the anterior part of the maxilla, diminished vertical development of the nasal cavity, and a retrognathically positioned maxilla due to decreased anteroposterior maxillary growth. This case report seems to indicate that the cartilaginous nasal septum is an important factor influencing vertical and sagittal growth of the maxilla. PMID- 9200256 TI - Anatomical guidelines for intranasal surgery of the lacrimal drainage system. AB - To facilitate identification of the nasolacrimal duct during intranasal surgery, we have determined the distances between the lacrimal drainage system and certain anatomical structures on the lateral wall of the nasal cavity. A total of 15 adult cadaver skulls were bisected mid-sagittally and evaluated morphometrically. In our specimens, the average distance from the natural ostium of the maxillary sinus to the nasolacrimal duct (NLD) was only 5.5 mm. This rather small distance should be taken into consideration, in order to prevent trauma of the NLD during surgical enlargement of the ostium of the maxillary sinus. The distances from NLD to the anterior surface of the bulla ethmoidalis, the free edge of the uncinate process and the attachment point of the middle turbinate on the lateral nasal wall were found to be 10.2 mm, 8.8 mm and 5.4 mm, respectively. Taking these distances into account, easy identification of the NLD during endonasal dacryocystorhinostomy surgery will be possible. PMID- 9200257 TI - Reduced nasal airway resistance following uvulopalatoplasty. AB - Active anterior rhinomanometry was performed on adult healthy snorers before and after uvulopalatopharyngoplasty or laser uvulopalatoplasty. Significant reduction of the nasal airway resistance both before and after pharmacological decongestion of the nasal mucosa was found in a group of 46 patients. Oedema disappearing after surgery may be an explanation for the results. PMID- 9200258 TI - Chemohormonal therapy for malignant melanomas of the nasal and paranasal mucosa. AB - We present three cases of primary malignant melanoma of the nasal or paranasal mucosa that were successfully treated by chemohormonal therapy using tamoxifen (TAM), an anti-estrogen agent. All of the patients showed good responses. TAM is widely known to be an anti-estrogen chemotherapeutic agent in the treatment of breast cancer and is thought to exert its anti-neoplastic effect in breast cancer tissues by competing with estrogen for estrogen receptors. The mechanism of the effect of TAM in malignant melanoma is not yet known. Although its anti neoplastic mechanism requires further exploration, we believe that chemohormonal therapy may become important in multidisciplinary treatment of malignant melanoma of the nasal and paranasal mucosa. PMID- 9200259 TI - Stereological estimation of blood vessel surface and volume densities in human normal and rhinitic nasal mucosa. AB - A technique is proposed for applying well-established stereological methods to study fixed nasal biopsy material to obtain an unbiased estimate of blood vessel surface and volume densities. Biopsies of the nasal mucosa from the anterior 10 mm of an inferior turbinate were obtained from 18 subjects (15 males, 3 females with a mean age of 28.5 years [range: 17-54 years]), ten of whom had perennial allergic rhinitis, and eight control subjects. The mucosal tissue volumes were estimated by water displacement. Zamboni's-fixed cryostat sections (10 microns thick), stained with haematoxylin and eosin, were examined histologically. Computerised images of randomly selected tissue sections were analysed with point counting and intercept-counting techniques to determine large blood vessel volume and surface densities, respectively. There were no significant differences between the volumes of tissue analysed from the control and rhinitic subjects (p = 0.35). The average volume density of the vessels was similar in the control group (6.17 +/- 1.41%) and the rhinitic group (7.8 +/- 5.59%; p = 0.38), but with a greater variability in the rhinitic group. Surface density estimations were 3.14 +/- 0.74 mm-1 in the control group and 3.10 +/- 1.41 mm-1 in the rhinitic group. Therefore, on average, the volume and surface densities of the cavernous blood vessels in rhinitis were unaltered and there was no evidence of vascular remodelling. PMID- 9200260 TI - Computerised tomography evaluation of the frontal recess in inflammatory diseases of the frontal sinus: standardisation of a new technique. AB - The authors present a new investigation technique by means of CT of the frontal sinus drainage pathway, the frontal recess, which could be of considerable help in defining its potential role in determining a chronic or recurrent inflammatory process of this cavity. Among the main characteristics of this technique are: (1) a clear presentation of the course and conformation of the recess and its relations with surrounding structures; (2) speedy, and therefore economical, operation (12 min for a complete examination); and (3) tolerability, because this examination starts off with axial-scans, which, compared to CT coronal projections and MRI scans, are less prone to defects and do not require strained postures. This this means that all kinds of patients can be assessed, which is a basic requirement for achieving standardisation. With axial scans the authors work from paraxial reconstructions (oblique sagittal) which, in their opinion, give the best definition of the frontal recess so far recorded. PMID- 9200261 TI - CO2 laser surgery of hypertrophied inferior turbinates. AB - The inferior turbinates are responsible for nasal obstruction in patients with allergic and vasomotor rhinitis. Until today there is no satisfactory means of treating hypertrophied turbinates. One hundred and eighty-four patients with nasal obstruction due to hypertrophied inferior turbinates were treated with the CO2 laser using the microscope and a micromanipulator. A few laser spots (1 W, 1 s, laser power density: 2,038 W/cm2) were applied to the head of the turbinate. After a few days a positive effect was present. One hundred and twelve patients were followed for over 2 years. Six months after laser surgery, 87.5% had excellent or good results. After one year, 82.1% of the patients were satisfied, and after 2 years 80.4% were satisfied. The procedure involves little bleeding, no pain and can be done under local anaesthesia in an outpatient setting. Less post-operative wound care is necessary. Therefore, the CO2 laser surgical technique can be considered as an effective method in the treatment of hyperthrophied turbinates. PMID- 9200262 TI - Septal reconstruction in nasal septum surgery with a composite-sandwich technique. AB - The authors describe a technique to reconstruct the medial and posterior portions of the nasal septum during surgical correction. This technique uses a "composite sandwich" made of two thin strips of Spongostan containing autologous crushed bone and cartilage. Thirty patients have been operated using this technique with satisfying results. PMID- 9200263 TI - Rhinolithiasis. AB - Rhinoliths are mineralized masses located in the nasal cavity. In this report, 12 patients with rhinolithiasis who were operated at the 2nd ENT Clinic of Ankara Numune Hospital are presented. The most frequently seen symptom is nasal obstruction, which has been seen in 9 patients. The disease most frequently seen in association with rhinolithiasis is chronic sinusitis. All masses have been extracted intranasally. PMID- 9200264 TI - A case of gustatory rhinorrhoea. AB - The authors describe a case of gustatory rhinorrhoea that appeared one year after skull trauma with delayed facial palsy. Traumatic interruption and abnormal regrowth of salivary parasympathetic fibers is hypothesized. In order to explain the pathogenesis of this syndrome an anatomical review of the transpetrosal nerves is included. A review of the literature is also presented. PMID- 9200265 TI - Endoscopic excision of a giant pyogenic granuloma of the nasal cavity caused by nasal packing. AB - A case of a giant pyogenic granuloma of the inferior turbinate secondary to nasal packing is presented and its removal via an endoscopic approach is detailed. The sinus endoscope provides excellent visualization and operative control during excision, obviating the need for a lateral rhinotomy. Pyogenic granulomas of the posterior nasal cavity are rare, and should be considered when a nasal mass is detected after packing for epistaxis. PMID- 9200266 TI - Von Graefe's circular saw for rhinoplasty. PMID- 9200267 TI - Recommendations for the reliable detection of illicit drugs in urine, with special attention to the workplace, in the European Union (December 1996). The Toxicology Experts' Working Group. PMID- 9200268 TI - Inhibition of nitric oxide synthase does not affect survival in a rat model of abdominal sepsis. AB - The effect of inhibiting nitric oxide (NO) synthesis during sepsis was studied in a caecal perforation model on Wistar rats. This model induces severe abdominal sepsis with a 48-h mortality > 90% in untreated animals. The survival time in hours (median values with the 95% confidence intervals in parentheses) for the control group was 15 (11.5-27.0) h. Treatment with the inhibitors of NO synthesis, NG-monomethyl-L-arginine (LNMMA), 30 mg kg-1 body weight (BW) or S-(2 aminoethyl)-isothiourea (AET), 3 mg kg-1 BW, given either once or twice after sepsis induction did not affect survival in this model. Survival time when LNMMA was given once was 13 (11-22) h and when given twice it was 14 (8-41) h. The corresponding survival times were 9 (4-28) h and 11 (5-27) h for treatment with AET. In conclusion, this study demonstrates that survival in the present model of live multiplying bacterial sepsis is not affected by either LNMMA or AET. Testing the potential clinical effects of inhibition of NO synthesis during sepsis should not be confined to short-term studies of haemodynamic changes induced by lipopolysaccharide. PMID- 9200269 TI - Sequential changes of inflammatory and nutritional markers in patients with community-acquired pneumonia. AB - The aim of the present study was to describe the long-term sequential changes of the acute phase proteins and of commonly used so-called nutritional markers in patients with community-acquired pneumonia (CAP), and to calculate the normalization rate of serum C-reactive protein (CRP), defined as the time for a 50% decrease, during the initial treatment of these patients. The long-term sequential changes of inflammatory and nutritional markers in patients with CAP have not been previously well-documented. However, in the diagnostic work-up of patients with suspected infectious diseases CRP levels are often used nowadays. Serum albumin, transthyretin (prealbumin), and transferrin together with serum iron, have often been used as "nutritional markers" in patients. We therefore studied the long-term changes of these parameters in patients with CAP, as these markers also are influenced by inflammatory reactions, in pneumonia for example. All the patients within the age range 50-85 years, with the exception of immunocompromised patients, who were admitted with CAP to the Department of Infectious Diseases at Danderyd Hospital during a 12-month period (January 1992 January 1993), were reviewed for inclusion in a prospective study of the long term sequential changes of inflammatory and nutritional markers in CAP patients. A total of 97 patients (50 men) with a mean age of 69.6 years were included in the study. Blood samples were drawn on admission, during the hospitalization period, and at the follow-up visits. Serum CRP, alpha 1-antitrypsin, haptoglobin and orosomucoid (alpha 1-acid glycoprotein) were used as acute phase proteins. However, albumin, transthyretin, and transferrin together with serum iron and percentage transferrin saturation were also included. Of all the parameters studied, CRP showed the greatest variation, already having the highest values at admission. CRP also showed, together with iron, the earliest response to recovery in the patients. The median time for a 50% decrease of CRP was 3.3 days for the patients (n = 73) with more than two CRP values measured during the first nine days. Transthyretin responded faster to patient recovery than did albumin. CRP showed the greatest amplitude of changes and together with iron and percentage saturation of transferrin it also showed the earliest response to recovery in patients with CAP. This indicates that CRP is the best of the parameters studied for use in diagnostic work-up and in follow up. PMID- 9200270 TI - The elimination of secretory leukocyte protease inhibitor (SLPI) from the gastrointestinal tract in man. AB - Secretory leukocyte protease inhibitor (SLPI) is the dominant protease inhibitor in the mucus secretions of the genital and respiratory tract and it was recently also detected in intestinal mucosa. Furthermore an earlier study showed high concentrations of SLPI in peritoneal fluid from patients with perforations of the large and small intestines. As SLPI is acid-stable, this raised the question of to what extent swallowed SLPI may contribute. The present study investigated the turnover of swallowed SLPI in the gastrointestinal tract. Native 125I-labelled SLPI was instilled in the duodenum of three healthy volunteers and radioactivity in plasma, faeces and urine was measured. Within 72 h 81.4% of the injected radioactivity was excreted in urine and 3.6% in faeces as radioactive low molecular weight proteins (< 1 kDa). Recombinant human SLPI (rh-SLPI) was incubated with porcine pepsin or human gastric or duodenal juice. This resulted in rapid degradation of SLPI to smaller molecules. In conclusion, SLPI is rapidly degraded in the stomach and duodenum. There were no measurable amounts of SLPI in the faeces. We have shown that during normal conditions, swallowed SLPI was rapidly degraded in the stomach and duodenum, and therefore it probably can not be of any importance for inflammatory diseases in the intestines. PMID- 9200272 TI - Serum and urinary markers of types I and III collagen turnover during short-term prednisolone treatment in healthy adults. AB - During recent years new sensitive serum and urinary makers have been introduced for assessment of collagen turnover. The aim of the present study was to assess whether short-term prednisolone treatment is associated with any adverse effects on serum levels of the type I collagen synthesis marker, the carboxy terminal propeptide of type I procollagen (PICP); on the type I collagen degradation marker in serum, the carboxy terminal pyridinoline cross-linked telopeptide of type I collagen (ICTP); on a serum marker of type III collagen synthesis, the aminoterminal propeptide of type III procollagen (PIIINP), or on the type I collagen degradation markers urinary pyridinoline (PYD) and deoxypyridinoline (DPD) concentrations. We studied 12 men and 8 premenopausal women aged 19-45 years (mean 31). All subjects were healthy. The design was a randomized double blind, placebo-controlled parallel group study with a 2-day run-in, a 3-day treatment period and a 4-day run-out. During run-in and run-out no medication was given. During the treatment period the subjects took either prednisolone, 40 mg per day, or placebo. Blood and urine were collected at the last day of each period. The intergroup comparisons of run-in treatment values showed that prednisolone suppressed PICP (p < 0.001) and PIIINP (p < 0.001). PICP levels remained suppressed during run-out, whereas PIIINP returned to pretreatment levels. NO prednisolone-induced effects on ICTP or on urinary PYD or DPD were detected by the intergroup comparisons. Short-term prednisolone treatment is associated with suppressive effects on type I and III collagen turnover. Whether serum PICP is more sensitive than urinary PYD and DPD for detection of short-term suppressive effects on type I collagen turnover remains to be further evaluated. PMID- 9200271 TI - Angiotensin-I-converting enzyme DD genotype is a risk factor of coronary artery disease. AB - Coronary artery disease (CAD) is a polygenic disease whose phenotypic manifestation depends on the interaction of a number of environmental factors. A number of genes, including the angiotensin-I-converting enzyme (ACE) gene, have been implicated in the pathogenesis of CAD. ACE could affect smooth muscle cell and fibroblast migration and proliferation, low-density lipoprotein (LDL) oxidation and endothelial cell function; these are all important factors in atherosclerosis. A polymorphic variant of the ACE gene correlates with higher circulating ACE levels and carries an increased risk of myocardial infarction, and cardiomyopathies. In this study, we sought to determine the distribution of ACE genotypes and the frequency of allele D in patients undergoing coronary angiography at our institution. DNA from 196 patients with angiographically proven CAD and 96 controls without CAD was amplified by polymerase chain reaction (PCR). The primers flanked the region of the ACE gene (intron 16) where the insertion (I) or deletion (D) of a 287-bp fragment results in the I/D polymorphism. PCR amplification of alleles I and D resulted in 490- and 190-bp products, respectively. In the control group, the relative allele frequencies of the polymorphism were similar to those of previously published European studies. The ACE genotype DD was present in 37.3% of patients with CAD as compared to 23.4% in the controls (p < 0.001, odds ratio 1.95, 95% confidence intervals (CI) 1.06-3.57). There was no association with the history of prior myocardial infarction. The genotype distribution in patients with single-vessel involvement was not significantly different from controls (p = 0.14). However, the DD genotype was significantly more common in patients having multivessel CAD when compared to single-vessel disease, indicating an association of this polymorphism with the extent of CAD. ACE genotype DD is more common in patients with multivessel CAD as compared to controls and to patients with single-vessel involvement, indicating that genotype DD is a genetic risk factor for extensive, multivessel CAD. PMID- 9200273 TI - Familial aggregation of LDL oxidation. AB - The "oxidation hypothesis" states that oxidative modification of low-density lipoprotein (LDL) is important in the pathogenesis of the atherosclerotic lesion. We studied 15 families (fathers, mothers and male twins of 16 to 18 years of age) to investigate the familial aggregation of LDL oxidation. As an indicator of LDL oxidation products we measured baseline levels of conjugated dienes extracted from LDL (LDL-BDC). For this analysis LDL was first isolated by rapid precipitation with buffered heparin. LDL-BDC was highest in fathers (mean 673 delta Abs per mg LDL cholesterol, 95% confidence interval (CI) 547-800) followed by mothers (500, 95% CI 408-592) and twins (383, 95% CI 337-430). There was a high correlation in the LDL-BDC between the identical twins (r = 0.81, 95% CI 0.44-0.95), but no correlation between the parents (r = -0.36). The LDL-BDC of boys correlated positively with that of fathers (r = 0.49, 95% CI 0.16-0.72), but not with that of mothers (r = 0.00). Highly significant positive correlations were observed between LDL-BDC and serum lipid risk factors among parents, but among twins the correlations were usually weaker. Our study suggests that inherited factors contribute to interindividual variability in the oxidative modification of LDL. PMID- 9200274 TI - Lowering of plasma phospholipid transfer protein activity by acute hyperglycaemia induced hyperinsulinaemia in healthy men. AB - Human plasma contains two lipid transfer proteins involved in the remodelling of plasma lipoproteins; cholesteryl ester transfer protein (CETP) and phospholipid transfer protein (PLTP). CETP mediates the transfer/exchange of cholesterylesters, triglycerides and phospholipids between high-density lipoproteins (HDL) and chylomicron (remnants), very low-density lipoproteins (VLDL) and low density lipoproteins (LDL). The physiological function of PLTP is unknown. It is able to transfer phospholipids (but not neutral lipids) between lipoproteins and to modulate HDL particle size in vitro. The effects of acute endogenous hyperinsulinaemia on plasma CETP and PLTP activity, as well as on lipid and lipoprotein levels, were assessed in eight healthy men during a 3-h hyperglycaemic clamp. Another group of seven men received an infusion of an equal volume of saline in order to detect possible dilution effects or effects on lipoprotein changes over time (control group). Plasma cholesterol and triglyceride concentrations fell during the clamp and the decreases were significantly different from the minor changes during saline infusion in the control group (p < 0.05 and p < 0.01, respectively). Plasma CETP activity levels did not change, but plasma PLTP activity levels decreased by 7.7 and 5.1% after 2 and 3 h of hyperglycaemia (p < 0.01 for each time-point). The hyperglycaemia induced mean percentage change in PLTP activity levels during the 3 h of the clamp was greater than the essentially absent change during the NaCl infusion (p < 0.05). Plasma PLTP activity during the clamp was related negatively to the insulin sensitivity index (p < 0.01 by analysis of covariance). It is concluded that acute hyperglycaemia-induced hyperinsulinaemia lowers plasma PLTP, but not CETP activity levels, either directly or in conjunction with an effect on plasma lipoproteins. PMID- 9200275 TI - Determination of the exocrine pancreatic function with the NBT-PABA test using a novel dual isotope technique and gas chromatography-mass spectrometry. AB - We describe a tubeless test of exocrine pancreatic function based on a new dual isotope technique, using N-benzoyl-L-tyrosyl-p-aminobenzoic acid (NBT-PABA) as a substrate for intestinal chymotrypsin activity and the stable isotope, 13C-PABA as marker. Gas chromatography-mass spectrometry (GC-MS) was used for the quantification of PABA and 13C-PABA in blood. The method involves hydrolysis, extractions, separation by HPLC, and methyl ester formation of the test substances before GC-MS analysis. The test is precise and shows good separation of healthy volunteers from patients with pancreatic insufficiency. The PABA/13C PABA ratios in serum after 1.5 h were 2.64 +/- 0.14 (mean +/- SEM) in 10 healthy volunteers and 1.26 +/- 0.22 in 10 patients with exocrine pancreatic insufficiency. We present a sensitive and specific assay, which is free of analytical interference and radiation hazards and, additionally, it illuminates extrapancreatic pharmacokinetic conditions. This test can eliminate the need for duodenal intubation, which makes it very acceptable to the patients. PMID- 9200276 TI - Serum and renal IGF-I levels after uninephrectomy in the rat. AB - The hypertrophy of the remaining kidney following uninephrectomy (UNx) has been related to an increase in renal insulin growth factor-I (IGF-I) content. However, while the increase in renal IGF-I lasts for only days after UNx, renal hypertrophy continues for months. In the present study we investigated whether IGF-I also plays a role in the late post uninephrectomy growth of the remaining kidney. Renal IGF-I in the remnant kidney was greater than that of control kidneys (78.3 +/- 17.3 vs 56.0 +/- 14.0 pmol g-1; p < 0.05) 3 days after UNx, tended to remain higher 30 days after UNx (83.8 +/- 23.6 vs 57.3 +/- 14.5 pmol g 1; p = 0.07), but was similar to that of the control kidney when examined 60 days after UNx (66.6 +/- 15.6 vs. 70.4 +/- 6.7 pmol g-1). Serum IGF-I in uninephrectomized rats was similar to that of controls 3 days after UNx, started to increase above the control level at day 10 after UNx and remained higher 30 and 60 days after UNx (75.9 +/- 6.9 vs. 48.7 +/- 7.3 nmol l-1 at 30 days, and 81.2 +/- 13.7 vs 52.9 +/- 11.0 nmol l-1 at day 60, p < 0.05 for both). The kidney weight of uninephrectomized rats was higher by 21% than that of controls 3 days after UNx, by 45% 30 days after UNx and by 63% 60 days after UNx (p < 0.05 for all three observations). At the end of the study, the glomerular volume of uninephrectomized rats was higher by 36% than that of the controls (p < 0.05) We suggest that in the rat, while the initial post uninephrectomy hypertrophy of the remnant kidney is associated with and most probably mediated by an increase in renal IGF-I, the hypertrophy that persists in later post UNx periods is associated with and may be mediated by an increase in serum IGF-I. PMID- 9200277 TI - Iodine nutrition in Sudan: determination of thyroid-stimulating hormone in filter paper blood samples. AB - In this study we examined the technique of measuring thyroid-stimulating hormone (TSH) on filter paper blood samples for use in evaluating the iodine nutrition status of newborns and adults living in iodine-deficient areas. Filter paper blood samples were obtained between the 5th and 7th day after birth from 103, 43 and 103 term newborns living in Khartoum (mild iodine deficiency), Kosti (moderate iodine deficiency) and Darfur (severe iodine deficiency), respectively. TSH was measured with a commercial assay and the levels were compared with those obtained with the same method in 1147 samples from term Swedish newborns, obtained on the 3rd to the 5th day of life. The mean (95% confidence interval) TSH levels of the three Sudanese groups and the Swedish group were 7.1 (4.8-9.4), 8.3 (6.6-10.1), 11.9 (0.9-22.9) and 4.51 (3.8-5.3) mU l-1, respectively. The mean TSH for all three Sudanese groups was higher than the Swedish mean (p < 0.001). TSH levels determined in filter paper blood samples from adults living in an iodine-deficient area showed a correlation (p < 0.001; r = 0.55) to levels in corresponding serum samples (range 0.52-14.1 mU l-1, median 3.4 mU l-1). No significant correlation was found, however, between blood spot levels and serum levels within the reference range (< 5 mU l-1). A modification of the commercial procedure consisting in using three instead of two monoclonal antibodies did not sufficiently improve the assay for measurements of TSH within the reference range. Thus, there is a need to develop the filter paper technique further to make it a useful test for monitoring iodine deficiency in populations expected to have TSH levels close to the reference range. In the examination of neonates, however, with their birth-induced surge of TSH, the current assay promises to be a convenient tool for discovering iodine deficiency within a community. PMID- 9200278 TI - The association between diabetic nephropathy and autonomic nerve function in type 1 diabetic patients. AB - Diabetic cardiovascular autonomic neuropathy increases the risk of deterioration in renal function and is associated with increased mortality in patients with renal failure. Type 1 diabetic patients with long diabetes duration, matched for age (38 +/- 9 years) and diabetes duration (28 +/- 8 years) were studied regarding the association between cardiovascular autonomic nerve function and different degrees of diabetic nephropathy. Eighteen patients were normo- (< 30 mg/l), six micro- (30-300 mg/l), and 13 macroalbuminuric (> 300 mg/l) based on urinary albumin concentrations in three separate morning samples. They were compared with 33 control subjects with similar age. Autonomic nerve function was evaluated by measuring the response of heart rate to deep breathing and active standing. Beat-to-beat finger artery blood pressure (Finapres) was tested during active standing. During deep breathing both change in heart rate (17 +/- 11, 9 +/ 7 and 4 +/- 3 beats/min) and ratio between expiratory and inspiratory R-R intervals (1.32 +/- 0.24, 1.14 +/- 0.15 and 1.05 +/- 0.04) decreased from normo- over micro- to macroalbuminuria (p < 0.05 vs normoalbuminuric and control subjects [17 +/- 5 beats/min and 1.28 +/- 0.10, respectively]). Similar results were obtained during active standing with respect to change in systolic arterial blood pressure (3 +/- 8, 2 +/- 13 and -6 +/- 11 mmHg; p < 0.05 vs control subjects [8 +/- 11 mmHg]). However, the response of diastolic arterial blood pressure or mean heart rate to standing up did not differ between any of the groups. The ratio of maximum to minimum R-R interval during the dynamic response of heart rate to active standing decreased with the degree of nephropathy (1.27 +/- 0.17, 1.11 +/- 0.11 and 1.05 +/- 0.06) with significantly higher values in patients with normo- compared with patients with macroalbuminuria (p < 0.05). All patients groups had significantly lower values than control subjects (1.46 +/- 0.22, p < 0.05). The overshoot of the blood pressure after an initial fall during active standing decreased with the degree of diabetic nephropathy. In conclusion, type 1 diabetic patients with long duration of diabetes have signs of cardiovascular autonomic neuropathy, the severity of which is related to the degree of nephropathy. PMID- 9200279 TI - Ranitidine effervescent and famotidine wafer in the relief of episodic symptoms of gastro-oesophageal reflux disease. AB - BACKGROUND: The aim of this study was to measure the efficacy of 150-mg ranitidine effervescent tablets compared with 20-mg famotidine wafers in the management of patients presenting to primary care physicians with episodic symptoms of gastro-oesophageal reflux disease (GORD). METHODS: The study was of a multicentre, open, randomized, parallel-group design in which 32 Norwegian general practitioners participated. After a pre-treatment demographic and symptom assessment, eligible patients were allocated to either ranitidine effervescent tablet or famotidine wafer. Patients were then provided with the study medication, a stopwatch, and a 2-week symptom diary card. Efficacy was primarily determined by the time to adequate symptom relief for the first symptom episode. RESULTS: In total, 377 patients were recruited to the study; 187 patients received ranitidine effervescent, and 190 received famotidine wafer. More than 50% of the patients had daily GORD symptoms before recruitment. Median time to adequate symptom relief was 15 min in the ranitidine group and 18.5 min in the famotidine group (P = 0.005). Adequate symptom relief within 60 min was reported by 165 (92%) ranitidine patients and 156 (84%) famotidine patients (P = 0.02). The number of non-responders after 60 min was twice as great in the famotidine group: 30 (16%) versus 15 (8%). A greater proportion of patients in the famotidine group liked taking the wafer formulation: 173 (94%) versus 126 (70%) (P = 0.001). CONCLUSION: There was a statistically significant difference in favour of 150-mg ranitidine effervescent tablets in terms of time to adequate symptom relief and the proportion of patients who achieved adequate symptom relief for the first episode. A greater proportion of patients in the famotidine group liked the type of formulation than in the ranitidine group. PMID- 9200280 TI - Evaluation of biologic gastrin activity of compound CI-988 in the isolated, vascularly perfused rat stomach. AB - BACKGROUND: The peptoid CI-988 has previously been shown to have high affinity for the cholecystokinin (CCK)-B/gastrin receptor and has been reported to be a powerful CCK antagonist in many systems, although it has agonist activity on histidine decarboxylase in the rat. METHODS: In the present study the effect of CI-988 on acid secretion and histamine release in the totally isolated, vascularly perfused rat stomach was assessed. RESULTS: CI-988 was found to be a gastrin agonist with regard to the stimulation of both histamine release and acid secretion. CONCLUSION: Thus, in this stomach model CI-988 behaved as a CCKB/gastrin agonist. The present study underlines the importance of testing the biologic activity of ligands in models with sufficient sensitivity. PMID- 9200281 TI - Peptic ulcer perforation before and after the introduction of H2-receptor blockers and proton pump inhibitors. AB - BACKGROUND: The aim of this retrospective study was to compare patients treated for perforated peptic ulcer before and after the introduction of the H2-receptor antagonists and proton pump inhibitors (PPI) with regard to their medical history, clinical features, methods of diagnosis and treatment, complications, and mortality. METHODS AND RESULTS: During the study period 1974 to 1992 we found a significant reduction in the incidence of peptic ulcer perforation (P < 0.001). Patients admitted during the later period of the study were older and more seriously ill. The incidence of perforation among men decreased, but that among women was stable, thus changing the sex ratio towards a female preponderance at the end of the study period. After the introduction of PPI the relative number of gastric perforations decreased compared with the number of perforations in the duodenum. A relatively higher proportion of patients with gastric perforations was taking acetylsalicylic acid or non-steroid, anti-inflammatory drugs at the time of admission compared with patients with duodenal perforation. Simple suture of the perforation was the operative procedure used in 80% of the patients. CONCLUSIONS: Even though patients were increasingly older and more ill, neither the mortality nor the rate of postoperative complications changed during the study period. PMID- 9200282 TI - Gastric juice levels of lactoferrin and Helicobacter pylori infection. AB - BACKGROUND: Recently, in vitro studies suggested that lactoferrin (Lf) might play an important role in the physiopathology of Helicobacter pylori-associated gastritis. However, whether Lf is present in the gastric juice and its relationship with H. pylori infection have not as yet been reported. In the present investigation the presence of Lf in gastric juice and its correlation with H. pylori infection were assessed. METHODS: This study comprised 30 H. pylori-positive and 14-negative patients with chronic gastritis. Gastric juice levels of Lf were measured with enzyme-linked immunoassays. Gastric juice concentration of Lf was also investigated in accordance with the histologic findings of biopsy specimens in the gastric body and antrum. RESULTS: Lf concentration in gastric juice was significantly higher in H. pylori-positive than in -negative patients (P = 0.033). The pH values are known to influence the levels of Lf. However, intragastric Lf levels were also significantly increased in H. pylori-positive patients as compared with H. pylori-negative patients after correcting the Lf levels for pH values (P = 0.029) or after adjusting the pH values of the gastric juice with NaHCO3 solution in both groups of patients (P = 0.0007). In addition, the gastric juice levels of Lf correlated significantly with the gastric mucosal concentrations of Lf in the gastric body (P < 0.005, r = 0.568) and the antrum (P < 0.05, r = 0.401). CONCLUSIONS: This study showed for the first time that Lf is present in gastric juice and that it correlates with H. pylori infection. Lf may constitute a good marker for H. pylori-associated gastritis. Although correlation does not prove causation, this study suggests that Lf might play an important role in the physiopathology of H. pylori associated gastritis. PMID- 9200283 TI - Eradication of Helicobacter pylori affects symptoms in non-ulcer dyspepsia. AB - BACKGROUND: The relationship between Helicobacter pylori infection and non-ulcer dyspepsia is controversial. METHODS: In a prospective, long-term, double-blind study we randomized 100 patients with non-ulcer dyspepsia and H. pylori infection to receive either of two treatment regimens: 1) bismuth-based triple therapy (n = 50) or 2) bismuth + placebo (n = 50). RESULTS: Triple therapy: subjects who became H. pylori-negative (n = 42) showed a significant symptomatic response when interviewed at 8 weeks, 6 months, and 1 year (P < 0.01). This improvement was evident in the 'ulcer-like' dyspepsia group at all times (P < 0.01) but in the 'reflux-like' and 'motility-like' groups at 6 months only (P < 0.01). Those who remained H. pylori-positive showed no decrease in symptoms at 8 weeks, 6 months, and 1 year. Bismuth-placebo therapy: subjects who became H. pylori-negative (n = 7) showed an improvement in symptoms at 8 weeks, 6 months, and 1 year. Those who continued to harbour the infection after treatment (n = 42) showed an insignificant improvement in the motility and non-specific groups only. CONCLUSION: This study shows that eradication of H. pylori results in a significant long-term reduction in symptoms of non-ulcer dyspepsia. PMID- 9200284 TI - Double-blind placebo-controlled study of cisapride in patients with nonspecific esophageal motility disorder accompanied by delayed esophageal transit. AB - BACKGROUND: Nonspecific esophageal motility disorder (NEMD) represents a difficult therapeutic challenge because of the heterogeneous nature of the esophageal motor functions. We studied the effects of cisapride on the esophageal symptoms and esophageal motor function in a group of patients with NEMD showing delayed esophageal transit. METHODS: Seventy eligible patients were entered into a 4-week, double-blind randomized comparison of 10 mg of cisapride or placebo, four times daily. Symptom assessment, esophageal manometry after wet swallows, and esophageal scintigraphy after intake of a liquid and solid bolus were performed in each patient before and after treatment. RESULTS: After 4 weeks of treatment cisapride significantly increased the prevalence of esophageal peristaltic contractions (percentage of total contractions, P < 0.05 versus base line and placebo) and significantly improved esophageal emptying of the solid bolus (P < 0.05 versus placebo) but not of the liquid bolus. Placebo did not have any significant effects versus base line on these variables. Both placebo and cisapride improved the distal esophageal amplitude versus base line (no significant intergroup differences). Symptom scores were significantly reduced after 4 weeks of treatment versus base line in both groups (no significant intergroup differences except for heartburn and regurgitation, P < 0.05). On global evaluation of treatment, significantly more patients in the cisapride group were rated as markedly or moderately improved, when compared with placebo. CONCLUSIONS: The results of the present study showed that cisapride is effective and well tolerated in patients with NEMD accompanied by delayed esophageal transit. Symptomatic improvement may possibly be related to its beneficial action on the esophageal body by increasing the number of peristaltic contractions and esophageal emptying of solids. PMID- 9200285 TI - Delayed gastric emptying during interferon-alpha therapy in patients with chronic hepatitis C: relief by cisapride. AB - BACKGROUND: Patients receiving interferon-alpha often experience symptoms such as upper abdominal discomfort, anorexia, and nausea, which suggest a delay in gastric emptying. Reduction of the dosages of interferon-alpha or even interruption of the treatment is sometimes required because of these symptoms. The present study was designed to investigate the effect of interferon-alpha on gastric emptying and to evaluate the effects of cisapride on gastric emptying and upper abdominal symptoms during interferon-alpha therapy. METHODS: Gastric emptying in 14 patients with chronic hepatitis C was estimated by the sulfamethizole capsule method before and 1 and 2 weeks after the beginning of interferon-alpha (6 million U/day) therapy. RESULTS: Before therapy none of the patients complained of upper abdominal symptoms, and all had normal gastric emptying. Interferon treatment delayed gastric emptying in 12 of the patients and induced discomfort and anorexia in 9 of the patients. The administration of cisapride reversed the delayed gastric emptying in six of seven patients and relieved abdominal discomfort and anorexia. CONCLUSIONS: These findings indicate that interferon-alpha delays gastric emptying and suggest that cisapride administration corrects the delayed gastric emptying and relieves the abdominal symptoms associated with interferon-alpha therapy. PMID- 9200286 TI - Glucagon-like peptide-1 7-36 amide and peptide YY have additive inhibitory effect on gastric acid secretion in man. AB - BACKGROUND: Glucagon-like peptide-1 7-36 amide (GLP-1) and peptide YY (PYY) are colocalized in the L-cell of the ileal mucosa, and both peptides may function as enterogastrone hormones. However, it is not known whether they interact with regard to the effect on acid secretion. METHODS: The effect of intravenous infusion of GLP-1 and PYY, either alone or in combination, on pentagastrin induced acid secretion in eight healthy volunteers was examined. The peptides were infused at two different rates: 0.25 pmol/kg/min (low rate) and 0.5 pmol/kg/min (high rate). RESULTS: Given alone, GLP-1 and PYY inhibited acid secretion by 26 +/- 5% and 18 +/- 5% (low rate) and 45 +/- 8% and 38 +/- 7% (high rate), respectively. Combined infusion resulted in an inhibition of 32 +/- 5% (low rate) and 62 +/- 7% (high rate). Both infusion rates resulted in GLP-1 and PYY plasma concentrations below or similar to postprandial levels. CONCLUSION: The present study suggests that the interaction between GLP-1 and PYY in man is of the additive type. The results indicate that GLP-1 and PYY have an important role in the physiologic control of gastric acid secretion. PMID- 9200287 TI - Luminal bacteria and small-intestinal permeability. AB - BACKGROUND: The influence of luminal bacteria on small-intestinal permeability has not been fully assessed. This study addressed this issue. METHODS: Thirty four subjects (mean age 64 years; range 22-95 years) were investigated for possible small-intestinal bacterial overgrowth (SIBO) with culture of a small intestinal aspirate. A lactulose/mannitol small-intestinal permeability test was performed, small-intestinal histology assessed and serum vitamin B12 concentrations measured in all subjects. Permeability was also assessed in a control group of 34 asymptomatic volunteers. RESULTS: Urinary lactulose/mannitol ratios were significantly increased in subjects with SIBO with colonic-type flora (P < 0.0005), even in the absence of villous atrophy. Urinary lactulose/mannitol ratios were increased in this group due to significantly increased urinary lactulose concentrations (P < 0.0005) rather than reduced urinary mannitol levels, after correcting for inter-subject variations in renal function. Counts of intraepithelial lymphocytes of CD8 phenotype were significantly increased in this group (P = 0.003). Although a significant correlation was found between intraepithelial lymphocyte counts and small-intestinal permeability overall (P < 0.002), these counts were not significantly different in subjects with SIBO with colonic-type flora whose permeability values were < or = > 0.028, the upper limit of normal in asymptomatic controls. Serum vitamin B12 concentrations did not differ significantly between groups (P > 0.5). Ageing did not independently influence small-intestinal permeability (P > 0.5). CONCLUSIONS: Small-intestinal permeability is increased in subjects with SIBO with colonic-type bacteria. This effect is independent of ageing and not mediated by vitamin B12 deficiency. Although counts of intraepithelial lymphocytes of CD8 phenotype are increased in this disorder, it is also unlikely that these cells play an important causative role in this process. Routine light microscopic assessment underestimates the prevalence of small-intestinal functional disturbance in this disorder. PMID- 9200288 TI - Evaluation of the role of neutrophils in the pathogenesis of acetic acid-induced colitis in mice. AB - BACKGROUND: Neutrophils are thought to play a role in the pathogenesis of inflammatory bowel diseases (IBD) such as ulcerative colitis and Crohn's disease, since prominent neutrophil infiltration has been observed in the inflamed colonic mucosa of patients with IBD. However, the role of neutrophils in the pathogenesis of IBD and experimental colitis remains equivocal. The aim of the present study is to clarify the possible role of neutrophils in the progression of acetic acid induced colitis in mice. METHODS: Using neutropenic mice treated with cyclophosphamide or with an LTB4 receptor antagonist, ONO-4057, the relationship between the severity of macroscopic colonic damage, the extent of myeloperoxidase (MPO) activities in the colonic tissues, and the number of neutrophils in the blood were examined after induction of colitis in mice. RESULTS: Changes of MPO activity in the colonic tissues paralleled well with the severity of the mucosal damage. In spite of a significant reduction in the number of neutrophils in the blood in cyclophosphamide-treated mice, neither the severity of mucosal damage in the colon nor the increase in MPO activities in the colonic tissues was affected 24 h after induction of colitis. Treatment with ONO-4057 significantly suppressed both the severity of mucosal damage in the colon and MPO activities in the colonic tissues in acetic acid-induced colitis in mice. CONCLUSIONS: The present results, obtained using treatment with cyclophosphamide and ONO-4057, show that the severity or the progression of acetic acid-induced colitis in mice was not influenced by a reduction of circulating neutrophils to about 25% of base line. PMID- 9200289 TI - Self-reported food intolerance in chronic inflammatory bowel disease. AB - BACKGROUND: Although suggested, it has never been convincingly documented that food sensitivity is of pathogenetic importance in chronic inflammatory bowel disease. However, many patients may relate their gastrointestinal symptoms to specific food items ingested and may restrict their diet accordingly. METHODS: A questionnaire was sent to all patients with chronic inflammatory bowel disease who attended the outpatient clinic, Medical Dept., Roskilde County Hospital in Koge, Denmark, in the year 1993. The patients were asked whether they had problems with any particular food item and, if so, to describe the symptoms experienced from it. A control group of 70 healthy persons were included. RESULTS: Among 189 patients, 132 (70%) responded. One hundred and thirty had completed the questionnaire, 52 males and 78 females aged 13-89 years (median, 43 years). Fifty-three (41%) had Crohn's disease (CD), 69 (53%) ulcerative colitis (UC), and 8 (6%) unclassified colitis. Forty-one patients (31 CD, 10 UC) were operated on; 51 (19 CD, 32 UC) had disease activity. Sixty-five per cent of the patients and 14% of the controls reported being intolerant to one or more food items (P < 0.0001). The intolerance covered a wide range of food products. The commonest symptoms among patients were diarrhoea, abdominal pain, and meteorism and among controls, regurgitation. Food intolerance was equally common in CD (66%) and UC (64%) and was not related to previous operation, disease activity or disease location. CONCLUSION: Most patients with chronic inflammatory bowel intolerance disease feel intolerant to different food items and may restrict their diet accordingly. The frequency and pattern of food intolerance did not differ between patients with CD and UC. The food intolerance was probably unspecific rather than of pathogenetic importance. PMID- 9200290 TI - Hereditary non-polyposis colorectal cancer: clinical features and survival. Results from the Danish HNPCC register. AB - BACKGROUND: Hereditary non-polyposis colorectal cancer (HNPCC) is a dominantly inherited syndrome characterized by the development of colorectal cancer (CRC) and other carcinomas. Our aim was to evaluate tumour parameters and survival in HNPCC. METHODS: One hundred and eight Danish HNPCC patients were compared with 870 patients with sporadic colorectal cancer. RESULTS: The median age at CRC diagnosis was 41 years in the HNPCC group. HNPCC patients had significantly more carcinomas located to the right colon (68% against 49% in controls), more synchromous tumours (7% versus 1%), more metachronous CRC after 10 years (29% versus 5%), more localized carcinomas (62% versus 39%), and significantly higher crude cumulative 5-year survival (56% versus 30%). CONCLUSIONS: CRC in HNPCC behaves differently compared to sporadic cases concerning age of onset, frequency of multiple lesions, and location. The metastatic tendency is less than in sporadic CRC and the survival is better. PMID- 9200291 TI - Interleukin-1 receptor antagonists and other markers in colorectal cancer patients. AB - BACKGROUND: Although the interleukin-1 receptor antagonist (IL-1ra) has been suggested as a potentially valuable therapeutic agent and has been shown to improve outcome in various animal models of arthritis, septic shock, and inflammatory bowel disease, there is little information available about its level in the circulation in patients with cancer. METHODS: Serum levels of IL-1ra, soluble interleukin-2 receptor (sIL-2r), soluble intercellular adhesion molecule 1 (sICAM-1), and cortisol were measured in normal controls and patients with colorectal cancer. RESULTS: The data showed that serum IL-1ra levels in patients were significantly lower than those of healthy controls (P < 0.05). In contrast, serum sIL-2r and cortisol levels in patients were significantly higher than those of normal controls (P < 0.01). Serum sICAM-1 levels in patients were the same as in normal controls. CONCLUSIONS: These results suggested that a reduced level of IL-1ra exists in colorectal cancer patients relative to normal controls, indicating that cancer patients have an immunologic disorder and that exogenous IL-1ra administration might be a future alternative for cancer treatment. PMID- 9200292 TI - Plasma YKL-40: a new potential marker of fibrosis in patients with alcoholic cirrhosis? AB - BACKGROUND: YKL-40 (human cartilage glycoprotein-39, or 38-kDa heparin-binding glycoprotein) is a mammalian member of a protein family that includes bacterial chitinases. YKL-40 mRNA is expressed by human liver and may play a role in tissue remodelling. The aims were to assess whether circulating YKL-40 is released or extracted in the hepatosplanchnic system and to localize YKL-40 in liver tissue. METHODS: Plasma YKL-40 was determined by radioimmunoassay in 25 patients with liver diseases (alcoholic cirrhosis (n = 20), chronic active hepatitis (n = 2), cirrhosis of unknown aetiology (n = 2), and fatty liver (n = 1) and in 18 subjects with normal liver function during a haemodynamic investigation with catheterization of liver vein and the femoral artery. Immunohistochemical studies of the localization of YKL-40 in cryostal liver biopsy specimens were obtained from eight other patients with alcoholic liver disease. RESULTS: Plasma YKL-40 was significantly increased in patients with alcoholic cirrhosis (median, 523 micrograms/l; P < 0.001) compared with controls (106 micrograms/l), and plasma YKL-40 in the hepatic vein was higher (P < 0.01) than that of the artery in both the patients and controls, showing release of YKL-40 from the hepatosplanchnic area. The release rate of YKL-40 from the hepatosplanchnic area was higher in patients with liver disease than in controls (11.0 versus 2.1 micrograms/min, P < 0.05). Furthermore, the highest plasma YKL-40 levels were found in patients with a moderate or severe degree of liver fibrosis, and immunohistochemical studies showed positive staining for YKL-40 antigen in areas of the liver biopsy with fibrosis. CONCLUSIONS: The increased plasma YKL-40 in patients with alcoholic cirrhosis may reflect the remodelling of liver fibrosis. PMID- 9200293 TI - Endogenous nitric oxide in exhaled air from patients with liver cirrhosis. AB - BACKGROUND: The aim of this study was to investigate the potential effects of liver insufficiency on nitric oxide concentrations in exhaled air. METHODS: Nitric oxide concentrations in the exhaled air from 13 patients with liver cirrhosis and 11 healthy control subjects was examined by the single-breath technique. RESULTS: There was a clear correlation between Child-Pugh score and NO in exhaled air (peak after 15 sec of breathholding, R = 0.623, P = 0.023). Similarly, there were significant correlations in peak NO concentrations and alkaline phosphatase, bilirubin, aspartate and alanine aminotransferase, and albumin. The most severely ill patient in our study had the highest NO concentrations in her exhaled air. On recovery from her liver illness the concentration of NO in her exhaled air decreased. There was no correlation between circulating levels of the endogenous NO synthase inhibitors asymmetric and symmetric N(G), N(G)-dimethyl-arginine and exhaled NO concentrations. CONCLUSIONS: The present data show a correlation between endogenous NO formation in the respiratory system and liver dysfunction. This might contribute to the understanding of the pathophysiology in pulmonary vasodilatation in liver disease. PMID- 9200294 TI - Treatment of small hepatocellular carcinoma associated with cirrhosis by percutaneous ethanol injection. A trial with a comparison group. AB - BACKGROUND: Ethanol injection has been reported to be effective in the treatment of hepatocellular carcinoma, but no controlled randomized trials have been performed. We therefore performed a trial comparing ethanol injection with an untreated, matched historical comparison group in the treatment of hepatocellular carcinoma. METHODS: From 1992 to 1993, 35 patients (14 Child's A and 21 Child's B cirrhosis) with small (< 4 cm) hepatocellular carcinoma associated with cirrhosis were treated by ethanol injection. Each patient was matched with an untreated case (followed up during the period 1984-89) for variables known to have independent prognostic value (age, Child's classification, number of lesions, alpha-fetoprotein, and modality of diagnosis). RESULTS: The 1-, 2-, and 3-year survival rates of ethanol-treated patients were 86% (95% confidence interval (CI), 69-94), 53% (95% CI, 34-68), and 33% (95% CI, 15-52), whereas the survival rates of the comparison group were 75% (95% CI, 56-85), 26% (95% CI, 13-41), and 14% (95% CI, 5-27) (P = 0.01). The 1-, 2-, and 3-year survival rates of Child's A were 100%. 87% (95% CI, 30-97), 71% (95 CI, 33-90), 71% (95% CI, 33-90) in the ethanol-treated patients and 92 (95% CI, 59-99), 43% (95% CI, 23-73), and 21% (95% CI, 23-72) in untreated patients. The 1-, 2-, and 3-year survival of Child's B patients were 76% (95% CI, 59-97), 32% (95% CI, 13-53), and 9% (95% CI, 0.8-33) in the treated group and 61% (95% CI, 40-83), 14% (95% CI, 3-32), and 9% (95% CI, 1-26) in the treated group. CONCLUSIONS: These data suggest that ethanol injection prolongs the life of patients with hepatocellular carcinoma associated with Child's A cirrhosis but seems not to influence the survival of Child's B patients. PMID- 9200295 TI - Hepatobiliary dysfunction and primary sclerosing cholangitis in patients with Crohn's disease. AB - BACKGROUND: Only a few studies have attempted to determined the prevalence of long-standing abnormal liver function and primary sclerosing cholangitis (PSC) in patients with Crohn's disease (CD). The aim of the study was to determine the prevalence of long-standing abnormal liver function test results and to describe the clinical, biochemical, and histologic findings in patients with large-duct classic PSC and small-duct PSC (that is, normal cholangiogram) in patients with CD during a 15-year period. METHODS: Patients with CD and long-standing abnormal liver function results were investigated individually with endoscopic retrograde cholangiography and liver biopsy. RESULTS: Of 262 consecutive patients with CD, 38 (15%) had long-standing increased alkaline phosphatase (ALP) values (mean, 1065 U/l; range, 321-4165 U/l). Of these, 10 patients were classified as having hepatic disease (4%), of which 9 had PSC and 1 had a non-specific reactive hepatitis. Of nine patients with PSC (3.4%), three were classified as having large-duct PSC; five, small-duct PSC; and one, unclassified. In patients with large-bowel CD (n = 102) the prevalence of PSC was 9%. Mean age at diagnosis of PSC was 35 years (22-46 years), and the female to male ratio, 7:2. All PSC patients had large-bowel involvement (P < 0.00015), and two of them developed colonic carcinoma of the large bowel (P < 0.01). All cases of small-duct PSC were stage 1, whereas large-duct PSC were stage 2-3. During the observation period (mean, 5.4 years) no PSC patients died. CONCLUSIONS: The results of our study indicate that PSC is the major hepatic disease in patients with CD and long standing abnormal liver function tests and is approximately as prevalent as in ulcerative colitis. Patients with PSC and CD may have a milder liver disease than patients with PSC and ulcerative colitis, perhaps because large-duct PSC is less common in patients with CD. Cholangiograms and liver biopsies are both needed to evaluate the extent of the disease. PMID- 9200296 TI - Randomization to surgery or observation in patients with symptomatic gallbladder stone disease. The problem of evidence-based medicine in clinical practice. AB - BACKGROUND: The description and understanding of gallbladder stone disease in the medical literature are difficult because an assessment of the natural course of this symptomatic disease, with separation of patients in strictly defined groups, is generally lacking. METHOD: A multicenter study was carried out with patients randomized to either surgery or conservative, expectant treatment to examine optimal treatment and natural history in well-defined groups of symptomatic gallbladder stone disease with pain, episodes only (study group 1) or acute cholecystitis (study group 2). The patients were between 18 and 80 years of age and had right upper quadrant or midline epigastric pain and ultrasonographic evidence of gallbladder stone, with or without acute cholecystitis. Medical treatment was ordinated on the basis of signs and symptom severity. Patients randomized to surgery were placed on the hospital's waiting list and electively operated on with cholecystectomy as soon as conveniently possible. Preliminary results of follow-up are based on questionnaires mailed at regular intervals and consultations if required by the patients' symptoms. Quality of life and freedom from pain are study end points for evaluation. RESULTS: Five hundred and one patients were considered for the study, 318 patients (63.5%) belonging to group 1 and 183 (36.5%) to group 2. One hundred and eighty-four patients (36.7%) were randomized, 120 (24%) in group 1 and 64 (12.7%) in group 2. Three hundred and seventeen patients (63.3%), with an equal proportion from each study group, were excluded, the largest number of patients because of incapacitating pain or serious presenting symptoms (26%). More women (81%) than men (19%) had pain alone (group 1), but the sex ratio was reduced, 56% women and 44% men, in patients with acute cholecystitis (group 2), with a highly significant difference (P = < 0.001) between the two groups. A significant difference in patient withdrawal from randomized treatment was registered, with 24% from observation and 12% from surgery (P = 0.032), but with an equal distribution in the two study groups. CONCLUSIONS: A randomized trial of this nature is feasible but extremely difficult to perform because of the heterogeneous nature of gallbladder stone disease, leading to exclusion of many patients and difficulties in measuring and evaluating outcome variables. PMID- 9200297 TI - Reversal of long-standing iron deficiency anaemia after eradication of Helicobacter pylori infection. AB - Helicobacter pylori has been proposed as a major determinant in multiple gastric disorders. We describe the case of a young adult with a long-standing medical history of sideropenic anaemia and of oral iron consumption dependence with a chronic superficial H. pylori-positive gastritis. All other causes of sideropenic anaemia were carefully excluded. Histology showed a peculiar pattern of non active H. pylori-positive gastritis. The bacterium was a non-VacA-producing strain. The first attempt at eradication caused a reduction in bacterial load and led to a partial normalization of haematologic variables without improving the ferritin level. A successful second course of eradication therapy completely reversed the anaemia and restored the iron deposit, which persisted at the 29 month follow-up. H. pylori infection can be involved in unexplained cases of iron deficiency anaemia in adults, and its cure can normalize the haematologic picture. PMID- 9200298 TI - Immature mucopeptic cell expansion in Helicobacter pylori-positive patients: a marker of progression towards gastric preneoplastic changes? PMID- 9200300 TI - The evaluation and treatment of functional constipation. AB - Constipation and defecation may be considered as the last taboo. The inability to defecate or to achieve this only by digital evacuation has never been a popular topic among patients and doctors. Application of tests from the colorectal laboratory has made it possible to study the function of the different parts of the colon and the mechanism of continence. We consider transit studies, defecography, EMG, and anal manometry, all useful as diagnostic procedures for functional constipation. Several causes of functional constipation can be distinguished in slow transit and difficult evacuation or colonic inertia, spastic pelvic floor syndrome, rectocele and intussusception. This article presents our view of the assessment and management of functional constipation. PMID- 9200299 TI - Hysterectomy: the anorectal pitfall. A guideline for evaluation. AB - Constipation following routine hysterectomy seems to occur more frequently than originally thought. Treatment depends on whether the patient is referred and to whom. Physical examination seems of limited value. Proper protocols for evaluation of complaints after hysterectomy are mandatory. Colonic transit studies and dynamic rectal examination could be useful. We found an overrepresentation of enteroceles in the hysterectomy group. Management of these abnormalities seems much more complicated than was previously thought. Prospective studies are needed to investigate anorectal disorders after hysterectomy. PMID- 9200301 TI - Gastro-oesophageal reflux in children. AB - Gastro-oesophageal reflux in children is different in several aspects from in adults. Pathophysiologically, 50% of reflux episodes are due to increased abdominal pressure which overcomes the lower oesophageal sphincter pressure. This pathophysiological abnormality disappears in children at the age of 1.5-2 years. Treatment is therefore different and aimed at thickening the gastric contents to inhibit reflux (Nutrition, Gaviscon, Algicon). The child is placed in the anti Trendelburg position when asleep. No further investigation or intensification of treatment is necessary in young children under the age of 2 years unless complications are present. With complicated gastro-oesophageal reflux, treatment in children is comparable to that in adults; the effects of H2 antagonists and proton-pump inhibitors are identical. Long-term complications of gastro oesophageal reflux are rare. In the near sudden death syndrome or acute life threatening events in infants due to total sphincter relaxation aspiration is possible and should be prevented. Optimal treatment and monitoring are mandatory. In mentally handicapped children rumination is more prominent than gastro oesophageal reflux. It is difficult to distinguish between vomiting, regurgitation and rumination. Treatment of oesophagitis might improve quality of life. When clear eosinophilic oesophagitis is observed food allergy should be considered and appropriately treated. PMID- 9200302 TI - The dumping syndrome. Current insights into pathophysiology, diagnosis and treatment. AB - The dumping syndrome is encountered in approximately 10% of patients after gastric surgery. A postprandial peripheral and splanchnic vasodilatation and ensuing relative hypovolaemia are pivotal in the pathophysiology of early systemic symptoms. Late dumping symptoms are a consequence of a reactive hypoglycaemia, which results from an exaggerated insulin and glucagon-like peptide-1 release. The diagnosis of dumping syndrome can reliably be made with the aid of a provocation test using 50 g glucose orally. Most patients with dumping can be treated with advice on diet and lifestyle. Octreotide effectively controls the signs and symptoms of dumping in patients refractory to standard therapy. It acts through its inhibitory effects on insulin and gut hormone release, a delay of intestinal transit time and inhibition of food-induced circulatory changes. Its long-term use is somewhat limited by side effects, particularly diarrhoea and steatorrhoea. PMID- 9200303 TI - Role of Helicobacter pylori in the pathogenesis of atrophic gastritis. AB - BACKGROUND: Atrophic gastritis is defined as a loss of the glandular structures and a collapse of the reticulin skeleton of the stomach mucosa. It is often accompanied by intestinal metaplasia. Both conditions result from long-term persistent chronic active gastritis and significantly increase the risk for gastric cancer. METHODS: Review of the role of Helicobacter pylori in the pathogenesis of atrophic gastritis. Specific attention is given to the classifications and histologic features of atrophic gastritis, the frequency with which atrophic gastritis occurs in H. pylori-infected subjects, the factors that influence the process of development of atrophic gastritis in the presence of infection, and the various mechanisms by which this bacterial infection may induce atrophic gastritis. In addition, the possible role of H. pylori in the etiology of auto-immune atrophic gastritis and pernicious anemia is discussed. CONCLUSIONS: H. pylori infection eventually causes atrophic gastritis in a considerable number of infected subjects. In different populations, the prevalence of atrophic gastritis increases by 1 to 3% per annum. Factors that may increase the risk for atrophy are infection at an early age, cytotoxin production by the infecting strain, and lowering of acid output. The association between H. pylori infection and the development of atrophic gastritis significantly supports the role of this infection in gastric carcinogenesis. PMID- 9200304 TI - Diagnosis of Helicobacter pylori infection. Review of diagnostic techniques and recommendations for their use in different clinical settings. AB - Multiple diagnostic tests are available for determining the presence of Helicobacter pylori infection. Some tests rely on endoscopy while others are non invasive. The accuracy of a test depends on the clinical situation in which it is used. Most tests have been evaluated in untreated individuals, while few studies have investigated their performance post-treatment when bacterial counts might be low. In this overview we first discuss the different diagnostic techniques available to the clinician, and their value in the pre-treatment and post treatment settings. Finally, we discuss the patient management-related question that arise in several clinical settings and recommend when to use which tests and why. PMID- 9200305 TI - New developments in Helicobacter pylori eradication therapy. AB - BACKGROUND: The optimal strategy for the eradication of Helicobacter pylori has yet to be determined. This paper summarizes some of the latest treatment strategies for eradicating H. pylori infection. METHOD: Literature review by author. RESULTS: Successful eradication of H. pylori requires the use of combination therapy involving control of gastric acid secretion together with anti-microbial drugs. The two most popular strategies use either a proton-pump inhibitor or an H2 antagonist plus two antibiotics-usually metronidazole, amoxycillin or clarithromycin. Ranitidine bismuth citrate is a co-precipitate of ranitidine hydrochloride and bismuth citrate, producing a high rate of H. pylori eradication when combined with clarithromycin. Future development of this combination may involve the addition of a second antibiotic. CONCLUSION: Modern combination therapy usually results in an 80-95% H. pylori eradication rate in compliant patients. PMID- 9200306 TI - Antiviral therapy of hepatitis C. AB - BACKGROUND: Chronic hepatitis C can be treated with interferon therapy, but persistent viral clearance is only achieved in 20% of patients. Which patients have a high chance of viral clearance and what other treatment might enhance the effectivity of interferon therapy are reviewed. METHODS: Data from published randomized trials on interferon mono-therapy, ribavirin mono-therapy and combination therapy of interferon-ribavirin and interferon-ursodeoxycholic acid are analysed separately and in a meta-analysis of individual data. RESULTS: Interferon mono-therapy leads to viral clearance in only 10% of patients with genotype 1 and in less than 10% in cirrhosis; patients with plasma HCV RNA detectable at 4 weeks of therapy have only 2% chance of viral clearance. Prolongation of therapy reduces relapse in treatment responders. Interferon ribavirin combination therapy appears to enhance the efficacy 2-3 fold without increasing toxicity. CONCLUSIONS: The benefit-risk/cost ratio of interferon mono therapy can be improved by selection of patients, monitoring plasma HCV RNA at 4 weeks, and prolonging therapy to 12 months in responders with genotype 1. Interferon-ribavirin combination is promising for its enhanced efficacy. PMID- 9200307 TI - Recent developments in screening, diagnosis and surgical treatment of hepatocellular carcinoma. AB - A relationship between the pathogenesis of hepatocellular carcinoma (HCC) and various types of underlying liver disease has been clearly demonstrated. Presently, this knowledge has not led to efficacious screening programmes for identifying patients with curable primary liver cancer at an early stage. Screening strategies should be improved in order to realize optimal management of patients with HCC. Detection and staging of primary liver cancer is hampered by the quality of imaging techniques and treatment is restricted by access to modalities such as liver transplantation. In this review recent developments in the management of hepatocellular carcinoma are discussed and an algorithm is proposed to define the most optimal approach for screening, diagnosis and surgical treatment. PMID- 9200308 TI - Acute liver failure: spontaneous recovery or transplantation? AB - BACKGROUND: Decision-making in acute liver failure. Acute liver failure is a disease with multiple organ involvement and a high mortality rate. Conservative management alone will only partly influence the outcome. The option of emergency liver transplantation has greatly improved survival rates, but unables spontaneous recovery. A set of prognostic criteria enables selection of patients who will benefit the most from emergency liver transplantation. METHODS: Retrospective review and survey of the Groningen results. RESULTS: Of 52 patients (33 adults and 19 children) admitted for acute liver failure 2 were beyond recovery and died, 9 were treated conservatively and recovered and 41 were listed for emergency liver transplantation because of an estimated survival rate < 20%. Of these, 3 died and 1 recovered spontaneously while waiting and 37 were transplanted. Survival rate for 41 patients listed for transplantation was 23 (56%) and was similar for children and adults. CONCLUSIONS: In patients with acute liver failure, management and decision-making in a specialized liver unit with the possibility of emergency liver transplantation is mandatory. PMID- 9200309 TI - An update on the pathogenesis and treatment of cholesterol gallstones. AB - The primum movens in cholesterol gallstone formation is hepatic cholesterol hypersecretion and chronic supersaturation of bile. A cascade of events will then include: (i) multiple biochemical defects: increased total biliary proteins (and qualitative shift to cholesterol crystallization-promoting factors), increased proportions of hydrophobic bile salts, increased mucin secretion, and rapid nucleation/crystallization of cholesterol from cholesterol-enriched biliary vesicles; (ii) multiple motility defects: impaired gallbladder contractility in vitro and gallbladder stasis in vivo, delayed intestinal transit. A genetic predisposition (together with environmental factors) might also be important. Therapy should be offered to patients with symptomatic gallstones. Cholecystectomy remains the only radical therapy for cholelithiasis. For a subgroup of patients with symptomatic, uncomplicated cholesterol stones who are unwilling to undergo surgery, or who have a significant surgical risk, alternative non-invasive therapies include: (i) oral litholysis of small stones by bile salts, (ii) fragmentation of 1-3 medium-sized stones by extracorporeal shock-wave lithotripsy, and (iii) topical dissolution of multiple stones by methyl tertbutyl ether. A major disadvantage of all non-surgical therapies, however, is the 50% recurrence rate of stones at 5 years. A number of prokinetic agents can improve gallbladder and/or intestine transit, two important contributing factors in gallstone disease. In selected patients, administration of these agents might enhance the clearance of cholesterol crystals/gallstones or might impede/delay gallstone formation and recurrence. PMID- 9200310 TI - Clinical implications of the sugar absorption test: intestinal permeability test to assess mucosal barrier function. AB - BACKGROUND: Functional integrity as an aspect of the mucosal barrier function of the small bowel can be estimated by the intestinal permeability for macromolecules. In the first part of this paper, an overview of intestinal permeability and its measurement is given. METHODS: In the second part of the paper our own experience with the Sugar Absorption Test using lactulose and mannitol to assess mucosal barrier function of gastric, small and large bowel, respectively, is described. RESULTS AND CONCLUSIONS: The Sugar Absorption Test is not recommended as a predictor of NSAID-related upper gastrointestinal damage nor as a marker of disease activity in inflammatory bowel diseases. The Sugar Absorption Test is very useful in screening for small intestinal disease, in assessing the response to treatment, and in predicting the prognosis, especially in coeliac disease. In our opinion, the D-xylose test is obsolete. PMID- 9200311 TI - Prevention of colorectal cancer. Costs and effectiveness of sigmoidoscopy. AB - There is increasing evidence that endoscopic polypectomy is a very effective means of preventing colorectal cancer. The feasibility of population-wide endoscopic screening is not established, however. The appearance of adenomatosis around the age of 60 in one-third of the population, many years prior to the manifestation of significant numbers of colorectal cancers, and the observation that the risk of developing metachronous adenomas correlates with pre-existing numbers of adenomas per colon suggest that a baseline endoscopy at 60 years with minimal follow-up may be a feasible screening strategy with a high rate of primary prevention in average risk individuals. Projections were made of the costs and benefits of various scenarios. A key element of this analysis was the assessment of probabilities to develop metachronous adenomas as a function of prior adenoma status by a mathematical approach using autopsy data. A screening strategy consisting of a baseline sigmoidoscopy at 60 years with follow-up restricted to 6% of the population was estimated to prevent 50% of colorectal cancers occurring after 60 years. A range of alternative scenarios, giving rates of primary prevention of colorectal cancer from 40 to 70%, had costs which were comparable to those of breast-cancer screening, but was far superior considering effectivity. PMID- 9200312 TI - Familial aggregation of inflammatory bowel disease: a population-based study in South Limburg, The Netherlands. The South Limburg IBD Study Group. AB - BACKGROUND: The aim of our study was to investigate the prevalence of Crohn's disease (CD) and ulcerative colitis (UC) in first-degree relatives of IBD patients living in a well-defined area. METHODS: IBD patients known at the IBD Registration South Limburg as well as population controls were asked about the occurrence of IBD in their first-degree relatives. RESULTS: IBD was reported and confirmed in 16 (out of 1554) relatives by 11 (out of 245) patients. Prevalence of IBD was highest for siblings (1.5%) and children (1.3%), while only 0.2% of the parents were affected with IBD. Among relatives of the control subjects, IBD was observed in 0.8% (versus 4.5% in IBD patients), resulting in an odds ratio of 5.7 (95% CI: 2.0-16.7). CONCLUSIONS: The observed risk of IBD for first-degree relatives of IBD patients was higher than in controls. However, the risk in our population is lower than has been reported by other centres, possibly because of the population-based character of our study. PMID- 9200313 TI - Mediators of mucosal inflammation: implications for therapy. AB - Treatment of inflammatory bowel disease remains a challenge. The major shortcoming in the development of new therapeutic approaches is the fact that the cause of inflammatory bowel disease is still unknown. Recognition of the importance of the arachidonic acid cascade of inflammatory mediators presents the opportunity to specifically inhibit or antagonize leukotriene B4, thromboxane, platelet activating factor, or phospholipase. Interleukins and cytokines have more recently been defined as targets for specific therapy. The results of these specific immune modulating studies are not only important from a therapeutic point of view, but substantially contribute to our understanding of the pathogenic cascades in IBD. In this review, several targets for novel therapeutic intervention are discussed. PMID- 9200314 TI - Role of animal models for the pathogenesis and treatment of inflammatory bowel disease. AB - The etiology of chronic inflammatory bowel diseases (IBD) such as Crohn's disease and ulcerative colitis is still unknown. Recent studies including animal models of intestinal inflammation have identified interactions between the mucosal immune system, host genetic susceptibility and environmental factors, including normal intestinal microflora. Although the ideal animal model of IBD has not been found, each model can study pathogenetic factors such as acute intestinal injury and healing, acute and chronic inflammation, regulation by key cytokines, T lymphocyte mediation, the role of luminal bacteria, immunoregulatory factors and genetic susceptibility. The relevance of these studies for the treatment of IBD is also discussed. PMID- 9200315 TI - New therapies for inflammatory bowel disease: an update on chimeric anti-TNF alpha antibodies and IL-10 therapy. AB - In the past few years new concepts have been formulated for the therapeutic management of difficult-to-treat inflammatory bowel disease, based on better insights in the pathophysiological processes in inflamed colonic mucosa. TNF alpha, for example, has been shown to play a major role orchestrating several other cytokines and pathways in the immunological network. TNF alpha is a pro inflammatory cytokine. In contrast, IL-10 is an anti-inflammatory cytokine. IL-10 deficient mice ('IL-10 knockout mice') will spontaneously develop enteritis in several parts of the digestive tract. Administration of IL-10 has been shown to improve and even to prevent the enteritis in these mice. One of the functions of IL-10 is to inhibit the TNF alpha production. Elimination of TNF alpha with anti TNF alpha antibodies and administration of IL-10 is, therefore, thought to achieve healing of the inflamed mucosa in man as well. The first uncontrolled studies with intravenous administration of chimeric anti-TNF alpha antibodies and with human recombinant IL-10 are hopeful for the future management of patients with inflammatory bowel disease; particularly since a fast healing of colonic mucosa was demonstrated and no serious adverse events were described. One has to be aware, however, of allergic reactions to the 'foreign' peptide. Because of rapid fibrosing and scarring in a narrow lumen, e.g. the ileum, stenosis formation could take place. In one described case a surgical intervention was needed because of this complication. For the near future these new and potent drugs seem very promising, although at present they are only available for trials with Crohn's disease patients. Certainly, in the coming years more immunomodulating drugs will be developed for use in man. PMID- 9200316 TI - Local corticosteroid injection in sport: review of literature and guidelines for treatment. AB - The risks and benefits of local injection therapy of overuse sports injuries with corticosteroids are reviewed here. Injection of corticosteroid inside the tendon has a deleterious effect on the tendon tissue and should be unanimously condemned. No reliable proof exists of the deleterious effects of peritendinous injections. Too many conclusions in the literature are based on poor scientific evidence and it is just the reiteration of a dogma if all steroid injections are abandoned. The corticosteroids represent an adjuvant treatment in the overall management of sports injuries: basic treatment is 'active' rest and graduated rehabilitation within the limits of pain. With proper indications there are only few and trivial complications that may occur with corticosteroid injections. Guidelines for proper local injection therapy with corticosteroids are given. PMID- 9200317 TI - Intra- and interobserver reproducibility of Cybex EDI 320 measuring spinal mobility. AB - The purpose of the present study was to conduct intra- and interobserver reproducibility tests of the measurement of spinal movement using an electronic digital inclinometer (Cybex EDI 320). Sixteen healthy subjects with no history of back pain were tested. Their mean age was 28.5 years (range 24-34). After palpation of reference points and a standard warming-up procedure, three repetitions of maximal ventral, dorsal, and lateral flexion were performed for measurements of lumbar and thoracolumbar movements. Intra- and interobserver reproducibility were conducted with 2-5 days interval. The coefficient of variance in the intraobserver test for ventral and lateral flexion varied between 4.3 and 10.1%, in the two interobserver tests between 3.4 and 13.8%. However, the coefficient of variance in intra- and interobserver tests for dorsiflexion varied between 18.1 and 27.6%. PMID- 9200318 TI - An experimental in vivo method for analysis of local deformation on tibia, with simultaneous measures of ground reaction forces, lower extremity muscle activity and joint motion. AB - This paper presents the pilot procedures of a new in vivo experimental method for measures of local bone deformation on tibia. The tibia transducer consists of a strain gauge mounted on a surgical staple, and was designed to measure local bone deformation. Pilot measurements were undertaken during two standardized conditions of forefoot and heel landing in seven healthy volunteers. Implantation of two tibia force transducers on tibia were performed under local anaesthesia. The local peak tibia deformation occurred at 20-42 ms (median) after ground contact, and was up to eight times higher during stance phase loading compared with standing still on one leg. Ground reaction forces, muscle activation patterns and kinematics were registered simultaneously, and were used to validate that the observed local deformation on tibia occurred under controlled and clinically relevant conditions. The new method may be used for investigating local deformation within various bone structures of the lower extremity. There are further methodological issues to address before major clinical interpretations may be concluded. In order to verify that the strain gauge transducer system was valid, a controlled displacement of the staple shanks was performed with a micrometer, and showed a linear relationship between applied deformation and strain gauge response (r = 0.97-0.99). In addition, a linear relationship was found between externally applied static forces and strain gauge response in a four-point bending cadaver system (r = 0.96-0.98). PMID- 9200319 TI - Effect of acute endurance and strength exercise on circulating calcium-regulating hormones and bone markers in young healthy males. AB - Physical activity plays a role in the maintenance of the skeleton but the mechanical, metabolic and hormonal mechanisms involved are largely unknown. The influence of acute endurance and strength exercise on circulating levels of calcitonin, parathyroid hormone (PTH), PTH-related peptide (PTHrP), osteocalcin, carboxyterminal cross-linked telopeptide of type I collagen (ICTP) and ionized calcium (Ca2+) was therefore evaluated. Eight healthy young males performed three exercise bouts on separate occasions: endurance exercise, i.e. cycling on a cycle ergometer for 45 min at 55% of Vo2max (E55%) and 15 min at 85% of Vo2max (E85%) and strength exercise at 85% of three repetitions maximum using a leg-press device (STR). Control experiments included the same subjects with the same time schedule but without exercise. Blood samples were taken before, immediately after exercise and during the recovery period. Hormones and bone markers were measured by use of various immunoassays. There was no obvious influence on calcitonin and PTHrP levels, whereas PTH was increased after strength exercise. ICTP and osteocalcin levels correlated positively at all times and showed regular variations. In comparison with the controls, ICTP levels showed a more pronounced decrease following physical activity whereas osteocalcin followed the same pattern as the controls except for after prolonged endurance exercise when a decrease was abolished. In conclusion, an increase in PTH after strength exercise and a pronounced decrease in ICTP after all exercise together with a relative increase in osteocalcin after prolonged endurance exercise might reflect some mechanisms involved in the positive effect of physical activity on bone mass. PMID- 9200320 TI - The effect of inhaled salbutamol and salmeterol on lung function and endurance performance in healthy well-trained athletes. AB - The present randomized, double-blind placebo-controlled study aimed at investigating the possible improvement in endurance performance caused by inhaled salmeterol (long-acting beta 2-agonist) and salbutamol (short-acting) compared to placebo in 18 healthy well-trained athletes, aged 17-30 years old. Lung function (flow-volume loops) was measured before and after each inhaled study drug and after run to exhaustion. After inhalation of study drug and 10 min warm-up, anaerobic threshold was measured; thereafter maximum oxygen uptake, peak ventilation and running time until exhaustion during a brief graded exercise were measured. No significant differences were found for ventilation, oxygen uptake or heart rate at anaerobic threshold or at maximum performance between placebo and the beta 2-agonists. Lung function increased significantly after exercise, but without differences between the beta 2-agonists and placebo. Running time till exhaustion was significantly reduced after both the long- and the short-acting beta 2-agonist compared to the placebo. PMID- 9200321 TI - Incidence of acute volleyball injuries: a prospective cohort study of injury mechanisms and risk factors. AB - The purpose of the study was to examine the incidence and mechanisms of acute volleyball injuries, with particular reference to possible risk factors for ankle injuries. Coaches and players in the top two divisions of the Norwegian Volleyball Federation were asked to keep records of exposure time and all acute volleyball injuries causing a player to miss at least one playing day during one season. We found 89 injuries among 272 players during 51588 player hours, 45837 h of training and 5751 h of match play. The total injury incidence was 1.7 +/- 0.2 per 1000 h of play, 1.5 +/- 0.2 during training and 3.5 +/- 0.8 during match play. The ankle (54%) was the most commonly injured region, followed by the lower back (11%), knee (8%), shoulder (8%) and fingers (7%). Of the ankle injuries, 79% were recurrences, and the relative risk of injury was 3.8 (P < 0.0001) for previously injured ankles (38 of 232) vs. non-injured ankles (10 of the 234). Moreover, a reinjury was observed in 21 of the 50 ankles that had suffered an ankle sprain within the last 6 months (42.0 +/- 7.0%; risk ratio: 9.8 vs. uninjured ankles; P < 0.000001). The data indicate that external supports should be worn for 6-12 months after an ankle sprain and that specific injury prevention programs may be developed for ankle sprains in volleyball. PMID- 9200322 TI - A twofold reduction in the incidence of acute ankle sprains in volleyball after the introduction of an injury prevention program: a prospective cohort study. AB - The purpose of this study was to examine the effects of an injury prevention program, consisting mainly of an injury awareness session, technical training (with emphasis on proper take-off and landing technique for blocking and attacking) and a balance board training program, for players with recurrent sprains. Baseline data were collected during the 1992-93 season and the program was introduced during the 1993-94 season. The 1994-95 season was used to evaluate the effects of the prevention program. The coaches and players in the top two division of the Norwegian Volleyball Federation kept monthly records of exposure time and acute injuries (causing a player to miss at least one playing day). The total exposure time was 149968 h, 132757 h of training and 17211 h of match play during the three seasons. The incidence of ankle injuries was reduced from 0.9 +/ 0.1 per 1000 player hours during the 1992-93 season (48 injuries) to 0.7 +/- 0.1 during the 1993-94 season (38 injuries; NS vs. 1992-93) and to 0.5 +/- 0.1 during the 1994-95 season (24 injuries, P < 0.01 vs. 1992-93). PMID- 9200323 TI - The treatment of acute soft tissue trauma in Danish emergency rooms. AB - Rest, ice, compression, elevation (RICE) is the most recommended treatment for acute traumatic soft tissue injuries. A questionnaire was given to all Danish emergency rooms (n = 5) regarding their routines for acute treatment of ankle sprains and muscle contusions. Complete answers were received from 37 emergency rooms (73%), covering the treatment of 111 ankle sprains and 101 muscle contusions. Treatment with RICE was given in a minority of injuries, ice (21%), compression (32%) and elevation (58%) similarly between injury types. A complete RICE treatment was rarely applied (3%). Verbal information on RICE and rehabilitation was given in less than half of the cases. We conclude that the acute treatment of ankle sprains and muscle contusions in the Danish emergency rooms is not applied in accordance with consensus from international literature, and that the instruction in rehabilitation should be improved. PMID- 9200324 TI - Immediate external compression in the management of an acute muscle injury. AB - In a prospective, non-randomized study 40 athletes with contusion or distension injuries to the thigh or the calf muscle were followed with tests of range of motion (ROM) of knee or ankle joint, test of serum creatine kinase (CK) and ultrasonography of the injury until completely recovered. An experimental group of 19 injuries where subjects received treatment with application of a maximum compression bandage within 5 min (mean = 2 min) of the injury was compared to a control group of 21 injuries where subjects were treated with rest and elevation only, and in some cases non-maximum compression after 10-30 min. No significant differences were noted with respect to time to complete subjective recovery, ultrasonic size of the injury or time to normal findings on ultrasound between treatment and control groups. Strain injuries, although showing a tendency to be smaller in size, took a longer time to complete recovery than contusion injuries (mean +/- SD = 26 +/- 22 days and 19 +/- 9 days, respectively, P = 0.02). Diagnostic CK values and reductions in ROM were not correlated to the severity of the trauma, while ROM showed weak correlation to the sonographically measured size of the hematoma (r = 0.42: P < 0.01). Injuries displaying a circumscript anechoic, low-echogenic or mixed lesion at the diagnostic ultrasound investigation normalized more slowly (P = 0.001) and took longer to complete recovery (P = 0.001) than injuries with diffuse hyperechogenic lesions. We conclude that in this study the application of a maximum compression bandage within 5 min of a muscle trauma did not significantly reduce the size of the hematoma nor significantly shorten the time to complete subjective recovery compared with no immediate treatment. The diagnostic ultrasound investigation was valuable in predicting the severity of the trauma. PMID- 9200325 TI - Conservative treatment of a partial Achilles tendon rupture with an intratendinous lesion. AB - The Achilles tendon is a common site of acute and overuse injuries in runners. A case is described here in which the diagnosis of a post-traumatic intratendinous lesion was based on clinical examination and magnetic resonance imaging (MRI), and where conservative treatment was given. After a 6 months follow-up, symptoms as well as the MRI verified that intra-tendinous structural abnormalities had disappeared. This case report demonstrates that conservative treatment may be sufficient to cure Achilles injury with severe structural changes inside the tendon. PMID- 9200326 TI - V(D)J recombination moves in vitro. AB - Substantial progress has been made recently in the study of the mechanism of V(D)J recombination, prompted in large part by the development of an in-vitro system for performing the DNA cleavage portion of the reaction. It is now clear that the RAG1 and RAG2 proteins play a direct role in DNA recognition and cleavage and that both RAG proteins are required for the nicking and strand transfer steps of the cleavage reaction. The heptamer of the recombination signal is critical for precise and efficient targeting of cleavage. In contrast, the nonamer plays an important role in initial sequence specific DNA binding and is contacted directly by RAG1. In-vitro systems have been established that perform cleavage at endogenous antigen receptor loci in intact nuclei, or that perform the complete recombination reaction, suggesting that progress in the field will continue at its current rapid pace. PMID- 9200327 TI - Accessibility and the developmental regulation of V(D)J recombination. AB - Antigen receptor genes are assembled from their component gene segments by a highly regulated series of site-specific DNA recombination reactions known as V(D)J recombination. Proteins encoded by the RAG1 and RAG2 genes are responsible for the recognition and double-stranded cleavage of a highly conserved DNA sequence flanking all rearranging gene segments. It remains uncertain how this common lymphoid recombinase is targeted to distinct loci in developing B and T cells and to specific loci at successive stages of lymphocyte development. This review considers evidence that DNA sequences which regulate the transcription of antigen receptor genes also regulate the recombination reaction by determining the accessibility of individual loci to the V(D)J recombinase. PMID- 9200328 TI - Events that regulate differentiation of alpha beta TCR+ and gamma delta TCR+ T cells from a common precursor. AB - T cells in vertebrates are composed of two distinct lineages of cells distinguished by the type of antigen receptors they express: the conventional population expresses TCR composed of alpha beta heterodimers while a smaller fraction of mature T cells displays gamma delta TCRs on the cell surface. The alpha beta and gamma delta T cells develop from common T-cell progenitors and the alpha beta/gamma delta lineage commitment process should be viewed in the context of the diverse cell fate decision processes crucial for normal cell differentiation from a single pluiripotent stem cell. A major focus in this field has been to determine the role of TCR gene rearrangement and expression in alpha beta/gamma delta lineage commitment. In this review we summarize the current state of understanding in the pattern of TCR gene rearrangement and expression of alpha beta and gamma delta T cells to address this central question of whether or not the lineage commitment is dictated by the state of TCR gene rearrangement. PMID- 9200329 TI - The RAD52 epistasis group in mammalian double strand break repair. AB - The S. cerevisiae RAD52 epistasis group gene products mediate DNA double strand break repair and recombination. These proteins and their modes of action have been extensively characterized. The existence of highly conserved mammalian RAD52 epistasis group homologues suggests that information regarding the functions and mechanisms of double strand break repair proteins in yeast may be applicable to mammalian recombinational DNA repair. Herein, we provide an overview of the S. cerevisiae RAD52 epistasis group and describe the characterization of the five mammalian RAD52 epistasis group homologues identified to date. In the context of their expression patterns and other functional analyses, we discuss potential roles for these proteins in mammalian recombinational DNA repair and specialized recombination events such as V(D)J recombination. PMID- 9200330 TI - The end-joining reaction in V(D)J recombination. AB - V(D)J recombination consists of a DNA cleavage reaction catalysed by RAG1 and RAG2, followed by an end-joining reaction that utilizes the cell's double-strand break repair machinery. Genes essential for the end-joining reaction include: XRCC4 encoding a protein of unknown enzymatic function; XRCC5 and XRCC6 encoding 86 and 70 kDa subunits of the Ku autoantigen, a DNA end-binding protein that is also the regulatory subunit of DNA-dependent protein kinase (DNA-PK); and XRCC7 encoding the catalytic subunit (DNA-PKcs) of DNA-PK. Recent progress in understanding the cleavage reaction, coupled with what was previously known about Ku, DNA-PK, and double-strand break repair, provide the foundation for a working model of how V(D)J recombination might be catalysed. PMID- 9200331 TI - Essential and perilous: V(D)J recombination and DNA damage checkpoints in lymphocyte precursors. AB - V(D)J recombination generates a diverse array of antigen-binding specificities, but breakage and re-joining of DNA segments have grave implications for the maintenance of genomic stability and oncogenic risk. Exposure of eukaryotic cells to genotoxic agents activates a DNA damage checkpoint that induces cell-cycle arrest and DNA repair, or apoptosis. We discuss several lines of evidence implicating DNA-dependent protein kinase (DNA-PK), and the gene mutated in ataxia telangiectasia (ATM), two mammalian homologues of yeast DNA damage-checkpoint genes, in regulating the response to intrinsic DNA damage that occurs during V(D)J recombination. PMID- 9200332 TI - Lymphocyte-specific genomic instability and risk of lymphoid malignancy. AB - Genetic instability is a force that links evolution, development and cancer. We have developed a quantifiable assay for a particular kind of lymphocyte-specific genetic instability (antigen receptor trans-rearrangements) mediated by the V(D)J recombinase complex. We find that the level of this type of instability correlates (at the population level) with risk for lymphoid malignancy. We have developed a murine model for this measure of instability to allow a more refined analysis of the genetic and environmental factors that sum to define population cancer risk. PMID- 9200333 TI - Synthesis and biological effects of N-alkylamine-labeled low-molecular-mass dermatan sulfate. AB - Dermatan sulfate (DS) is a component of connective tissue and catalyzes the heparin cofactor II-mediated inhibition of thrombin. Low-molecular-mass dermatan sulfates (LMMDS) are produced to prolong the antithrombotic activity of this substance. Cleavage of DS by nitrous acid leads to an LMMDS with a terminal 2,5 anhydrotalose (At) group at the reducing end which can react with primary amines. Tyramine (Tyr) was bound to the terminal At of LMMDS using reductive amination. LMMDS-tyr is produced using DS. LMMDS desacetglated were produced using totally deaminated DS. These compounds were employed as a model for the characterization of DS using NMR spectroscopy. The purity of the compounds was checked using capillary electrophoresis. The structure of the products was proven by 1H- and 13C-NMR spectroscopy. LMMDS-Tyr was radiolabeled with 125I for use in a radioimmunoassay. The anti-Xa activity and antithrombin activity of the tyramine labeled DS are very low. The clotting assays Heptest, aPTT, thrombin time, and ecarin time indicate a highly anticoagulant-active substance. The heparin cofactor II-mediated inhibition of thrombin is similar to the parent compound. LMMDS were labeled "endpoint-attached." They are a new tool to understand the actions of DS in biologic systems. PMID- 9200334 TI - Comparison of the pharmacodynamic and pharmacokinetic profiles of two low molecular-mass heparins in rats. AB - The pharmacodynamic and pharmacokinetic properties of certoparin and dalteparin were analyzed after intravenous and subcutaneous administration. The two different LMMHs exhibited different molecular mass and in vitro activities. The aim of the present study was to show the extent to which these in vitro differences influenced the biological activity and pharmacokinetics in vivo. It was possible to measure the plasma concentrations of the LMMHs by using a competitive assay with protamine-coated latex beads. Both LMMHs showed a biphasic aXa and aIIa activity curve after intravenous injection. Certoparin and dalteparin showed comparable aXa pharmacodynamics after IV and SC injection. By measuring the aIIa activities after IV administration of the LMMHs, T1/2, Amax, and AUC were comparable but tPC differed. After SC injection the aIIa activities of the LMMHs exhibited comparable T1/2 and Amax but different AUC and tPC. The plasma concentrations of the LMMHs showed comparable T1/2 but certoparin exhibited a higher Amax and AUC after IV and SC administration. The relative bioavailability of the aXa activity, aIIa activity, and the plasma concentrations ranged between 70 and 100%. The differences in the aIIa pharmacodynamic and pharmacokinetic profiles of certoparin and dalteparin may be caused by the differences in the in vitro activities and the different molecular mass. Clinical relevance of the different pharmacologic profiles is only expected of the aIIa activity-related biological effects of the LMMHs. PMID- 9200335 TI - Biochemical and pharmacologic characteristics of Reviparin, a low-molecular-mass heparin. AB - The introduction of low-molecular-mass heparins (LMMHs) has added a new dimension to the prophylactic and therapeutic management of thromboembolic disorders. These agents are now globally accepted as drugs of choice for postsurgical prophylaxis of deep vein thrombosis (DVT). Currently, the LMMHs are being developed for various therapeutic and cardiovascular indications. Reviparin is an optimized LMMH prepared by controlled nitrous acid digestion of porcine mucosal heparin. This drug has been developed utilizing validated procedures and exhibits a relatively narrow molecular mass distribution in contrast to most other commercially available LMMHs. The specific activity in the anticoagulant assays is approximately 40 U/mg whereas the specific activity in amidolytic anti-Xa assays is approximately 100 anti-Xa U/mg. Reviparin is capable of producing dose- and time-dependent antithrombotic effects in animal models of thrombosis. Although ex vivo anticoagulant effects are initially observed at dosages that are antithrombotic, this agent has been found to produce sustained antithrombotic effects when ex vivo anticoagulant actions are not measurable. Repeated administration of this LMMH induces progressively stronger antithrombotic effects. This drug has also been found to release tissue factor pathway inhibitor (TFPI) following both intravenous (IV) and subcutaneous (SC) administration. The studies included in this article are designed to provide additional data on the molecular profile using new calibration methods and additional results on pharmacologic studies. In particular, the release of TFPI following IV and SC administration to nonhuman primates is described. The effect of repeated administration of Reviparin mimicking the postsurgical prophylaxis of DVT is also reported in terms of any augmentation of the antithrombotic or hemorrhagic effects of this agent. PMID- 9200336 TI - Endothelial cell-associated tissue factor pathway inhibitor (TFPI) antigen in severe nondiabetic obese patients: effect of hyperinsulinemia. AB - It has been suggested that the extrinsic coagulation system plays a crucial role in the initiation of blood coagulation in atherosclerotic disease and that TFPI, the inhibitor of the factor VIIa/tissue factor complex, bound to the endothelial cells, could prevent in vivo blood clotting. Because obesity has a role in the pathogenesis of atherosclerotic cardiovascular disease, we measured TFPI antigen plasma levels (ng/mL) by ELISA at baseline and 5 minutes after an IV bolus of 20 IU/kg body weight of unfractionated commercial mucous heparin in 12 obese patients with a mean body mass index (BMI) of 41.4 +/- 1.4 kg/m2 and 14 normal weight control subjects (BMI 23.1 +/- 1.3 kg/m2). All subjects were submitted to an OGTT. The obese patients displayed a normal glucose tolerance. However, they had a different glucose-induced hyperinsulinemia (14.9 +/- 2.0 versus 7.8 +/- 0.8 mU/L, p < 0.01). Total serum cholesterol did not differ between controls and obese patients, whereas plasma triglycerides were higher in the latter group. Basal TFPI antigen plasma levels were similar in obese and controls (83.8 +/- 5.0 versus 77.7 +/- 3.5 ng/mL, p = N.S.). In contrast, after heparin a significantly lower rise in TFPI antigen plasma levels was observed in obese patients (511.2 +/ 43.4 ng/mL) (p < 0.003). Moreover, a significant inverse correlation was found between the heparin-stimulated TFPI antigen plasma levels and both BMI and basal plasma insulin concentrations. Thus, the link between insulin level and endothelial cell-associated TFPI could at least partially explain why obese patients are more prone to develop cardiovascular disorders. PMID- 9200337 TI - Tissue factor and plasminogen activator inhibitor type 2 expression in human stimulated monocytes is inhibited by heparin. AB - Stimulated monocytes are involved in blood clotting and fibrin dissolution by synthesizing tissue factor (TF) and fibrinolytic components such as plasminogen activator inhibitor type 2 (PAI-2). Heparin interacts with smooth muscle cells, platelets, and endothelial cells and specifically binds to human monocytes. In endothelial and smooth muscle cells, heparin selectively inhibits collagenase and tissue plasminogen activator gene expression. To investigate (1) heparin's influence on the hemostatic system by its interactions with plasma factors and cellular elements and (2) to determine its effects on gene expression in blood circulating cells, we studied the effect of heparin on TF and PAI-2 protein and mRNA in human lipopolysaccharide (LPS)- or interferon-gamma (IFN-gamma) stimulated monocytes. TF and PAI-2 proteins were investigated by ELISA and by assaying procoagulant activity. The mRNA study was carried out by an initial PCR screening followed by a Northern blot semiquantitative analysis. Heparin (0.5 U/mL) inhibited both TF and PAI-2 production and gene expression. The contemporaneous protein and mRNA decrease (TF and PAI-2 protein 22 and 42%, respectively; suggests that this action is, at least partially, at the transcriptional level. The effect is not specific for heparin and is not demonstrated by other glycosaminoglycans (chondroitin-4-sulfate or dermatan sulfate). This action may be relevant for the antithrombotic activity of heparin in cell-mediated blood clotting activation. PMID- 9200338 TI - Recombinant hirudin in the prevention of venous thromboembolism in patients undergoing elective hip surgery. AB - Venous thromboembolism is a major cause of mortality and morbidity in patients undergoing orthopedic surgery. In these patients systemic prophylaxis with anticoagulants appears to be the most effective approach to reduce venous thromboembolic events. Low-molecular-weight heparins (LMWHs) are the prophylactic agents of choice in patients undergoing elective hip replacement. However, their use is still associated with a 15% incidence of deep vein thrombosis (DVT); hence, there is a need for potentially more effective agents. Among these, hirudin and other selective thrombin inhibitors have recently been evaluated. The main pharmacologic properties of recombinant hirudin and the clinical trials carried out with this agent in the prevention of DVT in patients undergoing elective hip replacement are reviewed in this article. Overall, the results of these clinical trials are quite promising; they actually indicate that a b.i.d. subcutaneous injection of 15 or 20 mg of recombinant hirudin is an effective and safe prophylactic approach. Hirudin was more effective than unfractionated heparin, given at a low fixed dose, in two clinical trials and of a LMWH in one clinical trial. However, further studies are required to define the role of recombinant hirudin in the prevention of venous thromboembolism in orthopedic surgery patients. PMID- 9200339 TI - Prolonged prophylaxis in postoperative medicine. AB - Major surgical procedures, especially orthopedic surgeries, such as elective hip or knee replacement, are associated with a high incidence of postoperative deep vein thrombosis (DVT) and potentially fatal pulmonary embolism (PE). Although most surgeons exercise thromboprophylaxis, the length of time of prophylactic measures is at this time uncertain. With increasing shortening of in-hospital stays and thus shorter times of prophylaxis, increasing numbers of "late" PE are being recognized. These observations have raised the issue of postdischarge continuation of prophylaxis. This problem was recently addressed by four studies involving hip arthroplasty patients. In all trials prophylaxis was performed with low-molecular-weight heparins (LMWH). Two studies used enoxaparin, two used dalteparin. Duration of in-hospital treatment lasted from 7 to 15 days. The length of postdischarge prophylaxis ranged from 21 to 28 days. DVT was diagnosed by bilateral venography. In all instances there was a significant reduction in DVT in the treated patient group, compared with those who were not treated after discharge. It is assumed that this reduction also impacts the frequency of potentially fatal PE. The trials suggest that thrombosis prophylaxis should be continued in patients following discharge from hospital after major surgical procedures, especially when risk factors persist. PMID- 9200340 TI - Prophylaxis of venous thromboembolism in stroke patients. AB - Venous thromboembolism is a common complication in patients with acute thrombotic stroke. Estimates of the frequency of deep vein thrombosis (DVT) in untreated patients range from 20 to 75%. This wide range reported depends on the methods used to detect DVT and, importantly, on the degree of lower limb paralysis. Most thrombi occur in the paralyzed limbs in which the frequency ranges from 60 to 75%. Of these thrombi, 25% occur in the proximal segment and present a high risk for pulmonary embolism. Indeed, pulmonary embolism is the third most common cause of death in stroke patients and occurs in 1 to 2% of patients who do not receive prophylaxis. A number of methods of preventing DVT have been shown to be safe and effective in stroke patients. These include low-dose heparin, low-molecular weight heparin, and a heparinoid. Of these, the data with the heparinoid danaparoid provide the most solid evidence for efficacy, and in comparative trials it has been shown to be more effective than heparin. PMID- 9200341 TI - Evidence that endogenous heparin activity deficiency may be an important factor in atherogenesis. AB - Known atherosclerotic risk factors account today for only 50% of atherogenesis. Evidence is presented that a deficiency of endogenous heparin may account for the other half. Sensitive techniques have shown that there are trace quantities of heparin in plasma of humans. An inverse relationship has been found between plasma heparin levels and triglyceride-bearing Sf 12-400 lipoprotein, and a lower plasma heparin level could be an important determinant of atherogenesis fostering lipid abnormalities. Endogenous heparin also protects endothelium from harmful mediators that can impair normal function. Since atherosclerosis is a chronic inflammatory disease process, with monocyte-mediated release of cytokines and activation of integrins and phospholipase A2-mediated generation of platelet activating factor, heparin inhibits many of these events. Endogenous heparin activity thus opposes the effects of inflammatory activators. Heparin is also kown to inhibit complement activation and to suppress endothelin release from endothelial cells. In addition, endogenous heparin may suppress smooth muscle cell proliferation and decrease microthrombi formation on injured endothelial sites. All of these data seem to suggest that a deficiency of endogenous heparin or heparin-like substances predipose to atherosclerosis. It is conceivable that a genetically determined endogenous heparin deficiency is involved in atherosclerosis. PMID- 9200342 TI - Long-term anticoagulation of outpatients with adverse events to oral anticoagulants using low-molecular-weight heparin. AB - Bleeding complications are one of the major risks during oral anticoagulation. If further anticoagulation is indicated, low-molecular-weight heparin (LMWH) may offer an alternative treatment in those patients. In a prospective, nonrandomized study, 120 patients have been switched from oral anticoagulants to LMWH because of bleeding complications or other severe side effects during treatment with vitamin K antagonists. Indication for further anticoagulation was prophylaxis of recurrent thromboembolism, artificial heart valve replacement, atrial fibrillation with embolism and cardiomyopathy. The treatment period ranged from 2 months to 10.8 years. No fatal embolism occurred. One major but not severe episode of gastrointestinal bleeding occurred in a patient with an as yet unknown colon carcinoma. The cumulative treatment period amounts to 250 years. No drop in platelet count occurred in any patient. No other side effects were observed. LMWH was injected subcutaneously at doses ranging from 2500 to 15,000 anti-factor Xa units per day by the patient himself. The dose was adjusted on the basis of body weight, bleeding risk and thromboembolic risk. The results indicate that LMWH may be effectively and safely used as alternative anticoagulant regimen in patients with side effects or other complications on oral anticoagulants. PMID- 9200343 TI - Low-molecular-weight heparin therapy and mortality. AB - There is ample evidence from clinical trials to justify giving certain low molecular-weight heparins (LMWHs) subcutaneously rather than administering continuous intravenous unfractionated heparin for the initial treatment of venous thromboembolic disease. The LMWHs given by subcutaneous injection have a predictable anticoagulant response and prolonged duration of action. They can, therefore, be administered once or twice daily to treat venous thrombosis. Furthermore, treatment with these agents does not require laboratory monitoring. Eliminating the need for intravenous therapy and for laboratory monitoring should allow patients to be discharged earlier, and eventually lead to the outpatient treatment of venous thromboembolism. Studies to date indicate that LMWH is safer and as effective as continuous intravenous heparin in the treatment of venous thrombosis. The decreased mortality rates seen in two clinical trials, particularly in patients with metastatic cancer, were an unexpected but intriguing finding. This requires further confirmation, in larger prospective randomized trials. PMID- 9200344 TI - Treatment of deep vein thrombosis: is thrombosis regression a desirable endpoint? AB - Recent studies on the treatment of acute deep vein thrombosis (DVT) with low molecular-weight heparins have demonstrated that a certain degree of early recanalization of thrombosed veins can be obtained which is higher than that observed under standard heparin treatment. Thrombolytic treatment of DVT is mainly advocated because a reduction of late sequelae of DVT is expected from early thrombolysis. It has been made likely by several small long-term studies that this expectation is true, but conclusive evidence is still missing. There are also large differences in the reported incidence of late postthrombotic syndrome after acute DVT. It seems likely that there is a minimal reopening rate which is required to be of possible clinical value to the individual patient. A 30% or higher reduction of the Marder score is at present used in several clinical trials as a sign of individual response to the treatment and may prove to be a useful clinical endpoint in these and in future studies. Validated methods to predict the late sequelae of acute DVT, mainly severe postthrombotic syndrome, do not yet exist. Foot plethysmography, air plethysmography, duplex sonography (peak velocity of venous reflux, valve competence), and venous pressure reduction under exercise are possible candidates to be used in future prospective trials. From the existing evidence it is very likely that a marked or total reduction of thrombi will reduce the incidence of postthrombotic syndromes. Clinical studies aiming at a high rate of venous recanalization by prolonged treatment with low-molecular-weight heparins are ongoing. PMID- 9200345 TI - The natural history of deep-vein thrombosis. AB - Symptomatic deep vein thrombosis (DVT) carries a high risk for recurrent venous thromboembolism that persists for many years. This risk is dependent on readily identifiable risk factors, being low after a trauma or a surgical intervention, and higher in all the other patient categories. Severe post-thrombotic manifestations seem related with ipsilateral recurrence of DVT but are rare in patients with a first episode of venous thrombosis adequately treated with anticoagulant drugs and wearing compression elastic stockings. In the absence of malignancy, the prognosis with regard to survival is good. PMID- 9200346 TI - Anticoagulation in patients with heparin-induced thrombocytopenia type II. AB - Heparin-induced thrombocytopenia (HIT) together with simultaneously occurring thromboembolism is a serious complication of heparin treatment. At present an immunologic cause of this side effect of heparin is equivocally accepted. However, further anticoagulation of these patients is still debated. The present article summarizes the treatment of 20 patients with such complications. Two of these patients did not develop thrombocytopenia but presented cutaneous allergy or necrosis. All patients were treated either with intravenous heparinoid (Orgaran) or with low-molecular-weight heparin without/with simultaneous intravenous high-dose immunoglobulins or with intravenous r-hirudin. Based on these experiences the treatment of choice depends at present on the availability of the anticoagulants and on the local experience with the different anticoagulants. In the future r-hirudins and other nonheparin thrombin inhibitors may become the drugs of choice in this indication. Surgical intervention has to be considered additionally. PMID- 9200347 TI - Anticoagulation with Novastan (argatroban) in patients with heparin-induced thrombocytopenia and heparin-induced thrombocytopenia and thrombosis syndrome. AB - Heparin-induced thrombocytopenia (HIT) is a syndrome that has been identified with increased frequency. The mortality associated with HIT approaches 35%. Previous strategies for treatment of the associated thrombosis with HIT have frustrated clinicians with poor outcomes. Recent awareness of the complex pathophysiology of HIT combined with the availability of new anticoagulants has led to the development of a rational therapeutic strategy for this group of patients. The foundation of this strategy involves thrombin inhibition and careful patient monitoring. Our preliminary results with the thrombin inhibitor argatroban (Novastan) have been favorable and warrant continued investigation. PMID- 9200348 TI - Which glycosaminoglycans are suitable for antithrombogenic or athrombogenic coatings of biomaterials? Part I: Basic concepts of immobilized GAGs on partially cationized cellulose membrane. AB - Six different GAGs of different natural origin such as unfractionated heparin (HE), chondroitin sulfate (CS), dermatan sulfate (DS), keratan sulfate (KS), endothelial cell surface heparan sulfate (ESHS), and hyaluronan (HA) have been ionically immobilized onto partially cationized cellulose membranes with a substitution degree of 0.06. The GAGs have been characterized in terms of total sulfate content and relative molecular weight. The amount of immobilized GAGs was 10(3) times higher than the theoretical amount for monomolecular side-to-side coordination on polymer surfaces. In a standardized perfusion system with shear rates of 1050 sec-1, 37 degrees C, and 5 minutes with citrated blood, the level of platelet adhesion was 100% for partially cationized membrane, 90% for HA, 75% for each HE, CS, DS, KS, 5% for unmodified cellulose membrane, and 5% for confluent bovine aorta endothelial cells, referred to subendothelial matrix as standard for 100% platelet adhesion. ESHS-coated membranes were completely inert to platelets. The amount of ionically released GAGs during perfusion was also estimated. The partial cationic membrane is a suitable polymer surface in combination with the perfusion system to estimate the hemocompatibility of ionically immobilized water-soluble polyanions in terms of platelet adhesion at defined shear rates. The results of platelet adhesion are discussed in terms of structure and analytical parameter of the immobilized GAGs. PMID- 9200349 TI - Which glycosaminoglycans are suitable for antithrombogenic or athrombogenic coatings of biomaterials? Part II: Covalently immobilized endothelial cell surface heparan sulfate (ESHS) and heparin (HE) on synthetic polymers and results of animal experiments. AB - A systematic study was performed on immobilizing unfractionated heparin and endothelial cell surface heparan sulfate covalently with the spacer concept onto two polymer surfaces, followed by characterization of the surface concentration and in vitro and in vivo platelet adhesion properties under comparable high shear rates for microvascular vessels. Oligoamide spacer with a 16-atom chain length on cellulose surface and an 11-atom chain length on silicon surface, respectively, was used for immobilizing HE and ESHS via amino groups of glucosamine to the spacer which was anchored to the polymer surface. The surface concentration was in the range of 7 to 10 pmol/cm2 for HE and 1 to 1.5 pmol/cm2 for ESHS. This is in agreement with a calculated monolayer covering of ESHS and HE. In vitro and preliminary in vivo measurements (beagle, sheep) showed no platelet adhesion on the ESHS coatings, whereas HE showed high platelet adhesion and thrombus formation in vitro as well as in vivo. ESHS coating may be a potential candidate for preparing smooth artificial microvascular blood vessels. PMID- 9200350 TI - Anticoagulants and extracorporeal circuits. AB - Anticoagulants are pivotal to achieve circulation in extracorporeal circuits. In this review we discuss several anticoagulants in clinical use or in the preclinical phase. In hemodialysis the low-molecular-weight heparins (LMWHs) appear to be as effective and safe as standard heparin (SH). The main advantages of LMWHs in hemodialysis are the efficacy of a single loading dose, and the lack of laboratory control requirements. Newer anticoagulants such as dermatan sulfate and hirudin have been used in dose-finding studies in hemodialysis, although long term experience is lacking. LMW heparinoid may be used to replace SH or LMWH in the case of heparin-induced thrombocytopenia. In cardiopulmonary bypass surgery (CPB) heparin-coated extracorporeal circuits are now being commonly applied. Their main advantage is the requirement of lower systemic dosages of heparin. In CPB the place of LMWH or other anticoagulants needs to be investigated. PMID- 9200351 TI - Teratological studies on craniofacial malformations. AB - Craniofacial malformations cause great human suffering. The purpose of the experimental studies was to investigate teratogenically induced craniofacial malformations in the rat, and to study if vitamin B6 could prevent the teratogenically induced malformations in the rat. The aim of the clinical investigation was to compare mandibulofacial dysostosis (MFD) with hemifacial microsomia (HFM) and thalidomide-induced malformations restricted to the first and second branchial arches. In the experimental studies we used two different teratogenic agents, etretinate and BAPN (beta-aminoproprionitrile). Vitamin B6 was administered one day prior to and simultaneously with the teratogenic agent. The induced malformations were observed by direct microscopy, histology, differential staining, microdissection and enzyme histochemistry. Knowledge of isoenzymic differentiation was obtained by isoelectric focusing and polyacrylamide gel electrophoresis. The clinical features of 29 patients with MFD, 26 with HFM and seven with thalidomide-induced malformations were investigated. The patients underwent clinical investigations, radiography, tomography, computed tomography, surgical exploration and audiograms. The etretinate-induced syndrome in the rat shows similarities to first and second branchial arch syndromes in man. Defective formation of Meckel's cartilage and the cartilaginous skull base, the zygoma and the middle ear ossicles were prominent features of the observed malformations. The induced malformations were accompanied by increased staining for alkaline phosphatase (APase) in the skull and skull base cartilages and Meckel's cartilage. BAPN induced cleft palate in 95% of the cases and the teratogenically induced cleft palate was accompanied by a pathological differentiation pattern that could be traced by determination of isoenzymes in the palatal shelves as well as in amniotic fluid. Vitamin B6 could prevent the teratogenic malformations induced by etretinate and BAPN in the rat. Comparing MFD, HFM and thalidomide-induced malformations, all syndromes included patients with external, middle and inner ear malformations. Cranial nerve palsy/paresis was only seen in HFM and thalidomide-induced malformations. A relationship between disturbed neural crest cell migration and defects of the first and second branchial arches seems possible. PMID- 9200352 TI - Children's dental health in Europe. An epidemiological investigation of 5- and 12 year-old children from eight EU countries. AB - This thesis is based on a cross-sectional comparative study of dental health, treatment needs and attitudes to dental care in groups of 5- and 12-year-old children from the following eight cities in respective EU countries: Athens Greece, Berlin-Germany, Cork-Ireland, Dundee-Scotland, Gent-Belgium, Sassari Italy, Stockholm-Sweden and Valencia-Spain. A total of 3,200 children, 200 in each age group, were clinically examined by well-calibrated dentists, the parents completing a questionnaire on dental habits, parental and children's attitudes to dental care, smoking habits and parental occupations. The results disclosed pronounced differences in dental health and treatment need among the children from the different countries. The Scottish, Italian and German 5-year-olds exhibited the highest values for decayed, missing and filled teeth (dmft). The m component dominated for the Scottish sample, the d component in the Italian and d and f in the German sample. The highest values for DMFT in the 12-year-olds were found in the German, Greek and Italian samples followed by the Swedish sample. The F component dominated in the German and Swedish samples, while D dominated in the Greek and Italian samples. Analyses of the influence of socio-demographic and behavioural factors on the dental health, expressed as dmft/DMFT, showed that the most important factors explaining differences in caries experience were toothache, social class of the family and dental fear in the children. The frequency of similar attitudes (dental fear) in subjects and parents was 50% or higher in all the samples, and the frequency of similar dental attendance patterns in child and parent was 42% or higher in all the samples. For both age groups the proportion of subjects with regular dental attendance habits was highest in the Swedish, Belgian, German and Scottish samples. These findings, together with the high frequency of regular attenders without treatment need in the Swedish 5-year-olds indicate that organization of dental care must be closely adapted to the population it is set to serve. Separate strategies are necessary to manage the dental needs of healthy respectively diseased children. Reliable epidemiological data are necessary for planning, so that resources can be directed to the individuals with the greatest needs. However, to reach the children before onset of disease, parents, teachers, general health workers, sports coaches etc. must work jointly together with the dental profession. Among the eight countries, there is greater similarity in the organization of dental care for schoolchildren than for pre-school children. Only the Swedish system offers both preventive and restorative treatment irrespective of initiatives from the parents. In the other countries parents are mainly responsible for arranging for restorative treatment, above all for pre-school children. Different policies to promote dental health in the child population can be seen. Fluoridation of domestic water supplies has been implemented in Ireland, and the frequent use of fissure sealants in the Scottish, Irish and also the Belgian 12-year-olds is another example of a cost-effective measure influencing the dental health. PMID- 9200353 TI - My mother caused my illness: the story of a survivor of Munchausen by proxy syndrome. AB - OBJECTIVE: Munchausen by proxy syndrome (MBPS) is a form of child abuse in which a parent fabricates or produces illness in a child. Although the medical consequences of MBPS have been well described, there is no detailed published account of what it was like to grow up in a family where the mother systematically induced serious illness. This article describes one victim's childhood experiences. METHODS: The medical history was obtained from a review of the original medical records, notes from the primary physician, discussions with two physicians who provided treatment, and several meetings with the victim and the victim's therapist. RESULTS: This article chronicles the actual experiences of an MBPS victim through 8 years of medical abuse at the hands of her mother, reveals the victim's account of what happened to her, describes what her family was like, details the long-term consequences on emotional and physical development, identifies the factors that influence recovery, and details the impact on family relationships. CONCLUSIONS: Child maltreatment and MBPS need to be part of the differential diagnosis when the clinical picture is atypical or does not appear medically plausible. The consequences of MBPS are psychological and physical and impact the entire family. Suggestions to assist heath care providers recognize, assess, and report cases of suspected MBPS are provided. PMID- 9200354 TI - Trends in practice characteristics: analyses of 19 periodic surveys (1987-1992) of Fellows of the American Academy of Pediatrics. AB - OBJECTIVE: To examine 6 years of practice characteristics data of Fellows of the American Academy of Pediatrics (AAP), focusing on sex differences for specialty area, primary activity, practice setting, and practice location. METHODS: We analyzed data from 19 Periodic Surveys that were fielded between 1987 and 1992. The Periodic Survey is used to survey AAP members regularly about current issues in pediatric practice. There are no duplicate respondents in these analyses of the first 19 Periodic Surveys. We collapsed the 19 surveys into the years in which they were fielded, and analyzed sex differences for each of the 6 years. In addition, we ran logistic regressions on several questions, including all 16 868 respondents, to examine how the characteristics of the specialty have been affected by the increase in the number of female pediatricians, controlling for survey year, age of respondents, and specialty area practiced. RESULTS: The proportion of nonresident AAP members who are female has grown throughout the 6 years; in 1987, 26.9% were female, and in 1992, 36.4% were female. For 5 of the 6 years there were sex differences in specialty area, usually concerning pediatric subspecialties. Substantial sex differences occurred in primary activity, in which each year women were more likely than men to be salaried. Men were more often in group practices, whereas women were generally more likely to practice in hospitals or clinics. Logistic regression demonstrated that there are sex differences in practice characteristics across time, but there is also a substantial change in practice characteristics accountable to survey year, eg, a pediatrician of either sex was 75% more likely to be salaried in 1992 than in 1987. CONCLUSIONS: Throughout the 6-year period, AAP members became increasingly more likely to practice general pediatrics, to be salaried, and to be younger-all effects independent of sex, all effects stronger for females. Rapid transformations in the health care system will likely reduce current sex differences in practice characteristics of the future. PMID- 9200355 TI - Mycobacterium tuberculosis transmission from a pediatrician to patients. AB - The following report describes the contact investigation of a pediatrician with tuberculosis (TB). The pediatrician's disease was discovered in late February 1993 after tuberculin skin testing (TST) of his 15-month-old son was positive (13 mm induration). Further investigation to identify the source of the child's infection revealed a positive (15-mm induration) TST in the pediatrician. The pediatrician had been symptomatic with a cough since September 1992. The pediatrician had a chest radiograph that revealed numerous cavitary lesions and a sputum smear that was positive for acid-fast bacilli. An investigation was initiated to assess whether the transmission of Mycobacterium tuberculosis had occurred in the pediatrician's office to patients, families, or other visitors. The investigation was later extended to include the hospitals and the day care center where the pediatrician worked. METHODS: A letter was mailed to parents of children served by the practice, explaining the potential exposure to TB and requesting that all persons who visited the office after September 1, 1992 complete an interview and Mantoux TST. Mass interviewing, testing, and test interpretation within the practice took place seven times during March and April 1993. RESULTS: At the completion of screening, 181 (87%) of 208 children who had close contact with the index case were reliably skin-tested and returned for interpretation. Three (1.7%) of the 181 children were TST-positive (>/=5 mm). Thirty-seven (13%) of the 286 adults tested and returning for interpretations were TST-positive (>/=10 mm). Thirty-two (86%) of the 37 adults who tested positive were foreign-born. CONCLUSION: This investigation highlighted the need for identifying childhood TB infection as a sentinel event for adult disease. It also demonstrated the difficulty associated with deciding the extent of contact investigation of a health care worker with TB. Finally, the investigation emphasized the importance of maintaining regularly scheduled and appropriate testing for TB infection in health care workers and the need for health care workers to be cognizant of their own risk and be able to identify, especially in themselves, signs and symptoms of potential TB disease. PMID- 9200356 TI - Safety and pharmacokinetics of multiple doses of recombinant human CuZn superoxide dismutase administered intratracheally to premature neonates with respiratory distress syndrome. AB - OBJECTIVES: To examine the safety and pharmacokinetics of multiple intratracheal (IT) doses of recombinant human CuZn superoxide dismutase (rhSOD) in premature infants with respiratory distress syndrome who are at risk for developing bronchopulmonary dysplasia (BPD). Methods. Thirty-three infants (700 to 1300 g) were randomized and blindly received saline, 2.5 mg/kg or 5 mg/kg rhSOD IT within 2 hours of surfactant administration. Infants were treated every 48 hours (as long as endotracheal intubation was required) up to 7 doses. Serial blood and urine studies, chest radiographs, neurosonograms, SOD concentration and activity measurements, and tracheal aspirate (TA) inflammatory markers were assessed throughout the 28-day study. RESULTS: SOD concentrations in serum (0.1 [0.05/0.15] microg/mL-geometric mean with lower/upper confidence intervals), tracheal aspirates (TA) (0.2 [0.1/0.3] microg/mL) and urine (0.3 [0.2/0.4] microg/mL) were similar at baseline in all 3 groups and did not change significantly in the placebo group. In the rhSOD treatment groups, SOD concentrations were increased on day 3 and did not change significantly thereafter over the 14-day dosing period (also measured on days 5, 7, and 13). SOD concentrations averaged 0.4 [0.3/0.5] microg/mL in serum, 0.8 [0.6/1.2] microg/mL in TA and 1.1 [1.0/1.3] microg/mL in urine for the low-dose group and 0.6 [0.5/0.7] microg/mL in serum, 1.1 [0.9/1.5] microg/mL in TA, and 2.2 [1.6/2.9] microg/mL in urine for the high-dose group over the 14-day dosing period. Enzyme activity directly correlated with SOD concentration and rhSOD was active even when excreted in urine. TA markers of acute lung injury (neutrophil chemotactic activity, albumin concentration) were lower in the rhSOD agroups compared with placebo. No significant differences in any clinical outcome variable were noted between groups. CONCLUSIONS: These data indicate that multiple IT doses of rhSOD increase the concentration and activity of the enzyme in serum, TA and urine, reduce TA lung injury markers and are well-tolerated. Further clinical trials examining the efficacy of rhSOD in the prevention of BPD are warranted. PMID- 9200357 TI - Comparison of Infasurf (calf lung surfactant extract) to Survanta (Beractant) in the treatment and prevention of respiratory distress syndrome. AB - OBJECTIVE: To compare the relative safety and efficacy of Infasurf (calf lung surfactant extract; ONY, Inc, Amherst, NY, IND #27169) versus Survanta (Beractant, Ross Laboratories, Columbus, OH) in reducing the acute severity of respiratory distress syndrome (RDS) when given at birth and to infants with established RDS. DESIGN: A prospective, randomized, double-blind, multicenter clinical trial. SETTING: Thirteen neonatal intensive care units participated in the treatment arm: seven of these concurrently participated in the prevention arm. PATIENTS: The treatment arm enrolled infants of /=48 hours. Differences were significant in July and November 1995, and in the final survey in March 1996. Nevertheless, in March 1996, 38% of short stay infants were scheduled to be seen 4 or more days after discharge, and 33% 14 days after discharge. CONCLUSION: Although follow-up practices have changed in response to shorter newborn hospital stays, a significant proportion of pediatricians are not following the American Academy of Pediatrics guidelines for the follow-up of short-stay infants. Whether or not failure to follow these guidelines will lead to an increase in morbidity is unknown. PMID- 9200363 TI - Infant sleep position in licensed child care centers. AB - OBJECTIVE: To determine 1) familiarity of child care centers with American Academy of Pediatrics (AAP) recommendations regarding infant sleep position, 2) predominant infant sleep positions in child care settings, and 3) child care policies pertaining to sleep position for infants less than 6 months of age. DESIGN: A descriptive, cross-sectional telephone survey including the age and number of infants cared for, infant sleep positions currently in use, and details regarding reasons for sleep position policies. PARTICIPANTS: All licensed child care centers caring for infants less than 6 months of age in Washington, DC, and Montgomery and Prince Georges Counties in Maryland. RESULTS: Out of 137 centers in these areas that accept infants less than 6 months of age, 131 completed the survey. Only 57% (75) of the centers were aware of recommendations regarding infant sleep position. Infants were placed prone in 49% (64) of the centers and 20% (26) of the centers positioned infants exclusively in the prone position. Of the centers, 75% (98) did not have a written policy regarding sleep position. Most common reasons for placing infants in the prone position included child comfort, fear of choking, and guidance by the parents of the infants. Centers that used the prone position exclusively cared for significantly fewer infants on average than centers that never or only sometimes placed infants prone. CONCLUSIONS: Almost half (43%) of licensed child care centers surveyed in the greater Washington, DC area were unaware of the association between sudden infant death syndrome (SIDS) and infant sleep position. Child care centers aware of prone positioning as a SIDS risk were less likely to place infants to sleep in this position, with many such centers avoiding prone positioning entirely. However, it was common for centers aware of the SIDS risk to still place infants prone if directed to do so by parents or if concerned about child comfort. Further educational efforts directed toward child care providers are needed. PMID- 9200364 TI - Mortality within the first 2 years in infants exposed to cocaine, opiate, or cannabinoid during gestation. AB - OBJECTIVE: To determine the mortality rate, during the first 2 years of life, in infants who were exposed to cocaine, opiate, or cannabinoid during gestation. METHODS: For a period of 11 months, a large group of infants were enrolled and screened at birth for exposure to cocaine, opiate, or cannabinoid by meconium analysis. Death outcome, within the first 2 years after birth, was determined in this group of infants using the death registry of the Michigan Department of Public Health. RESULTS: A total of 2964 infants was studied. At birth, 44% of the infants tested positive for drugs: 30. 5% positive for cocaine, 20.2% for opiate, and 11.4% for cannabinoids. Compared to the drug negative group, a significantly higher percentage (P < .05) of the drug positive infants had lower weight and smaller head circumference and length at birth and a higher percent of their mothers were single, multigravid, multiparous, and had little to no prenatal care. Within the first 2 years of life, 44 infants died: 26 were drug negative (15.7 deaths per 1000 live births) and 18 were drug positive (13.7 deaths per 1000 live births). The mortality rate among cocaine, opiate, or cannabinoid positive infants were 17.7, 18.4, and 8.9 per 1000 live births, respectively. Among infants with birth weight /=85th percentile. More then 30% of NIDDM youths presented with hypertension. Diabetic ketoacidosis was present in >25% of NIDDM patients. Acanthosis nigricans was documented in 86% of NIDDM and 0% of IDDM patients. CONCLUSIONS: In Arkansas, youths with NIDDM are characterized by significant obesity in contrast to youths with IDDM. Physical characteristics such as obesity, acanthosis nigricans, and hypertension on examination of any youth with new-onset diabetes should raise suspicion of NIDDM. PMID- 9200366 TI - Determinants of behavior in homeless and low-income housed preschool children. AB - OBJECTIVES: To describe the characteristics of homeless and low-income preschool aged children, and to identify family and environmental determinants of their behavior. METHODS: An unmatched case-control design was used to recruit a sample of sheltered homeless families and a comparison group of low-income housed families who were never homeless in Worcester, Massachusetts. Seventy-seven sheltered homeless and 90 low-income housed mothers with preschool-age children were assessed using a comprehensive interview protocol. Information about mothers' housing, income, service use, par-enting practices, and children's father was obtained. Data about children's background, health, and life events were included. Standardized instruments were administered to assess mothers' mental health and their children's behavior. Comparisons of homeless and low income housed families were used to describe the sample of 167 preschoolers. Multiple linear regression was used to examine the association of various stressors, such as homelessness, and family factors with their behavior. RESULTS: Although homeless preschoolers were significantly more likely to have experienced stressful life events, undergone a care and protection investigation, and been placed in foster care when compared with low income preschoolers, differences in adverse behaviors were minimal. Although homeless children scored higher than housed children on the internalizing, externalizing, and total problem score on the Child Behavior Checklist (CBCL) (52.5 vs 49.9, 54.8 vs 51.2, and 54.4 vs 51.1, respectively), approximately equal numbers of children from both groups scored in the clinical range. With regard to determinants of behavior, mothers' emotional status was one of the strongest independent predictors of negative behavioral outcomes on both subscales. Foster care placement and death of a child's friend were predictors of adverse internalizing behavioral outcomes on the CBCL. After controlling for housing status, parenting practices, child's age, child's history of physical abuse, and specific life stressors predicted adverse externalizing behavioral outcomes. For both subscales, housing status and behavior were only marginally associated in the multivariate model. CONCLUSIONS: Both homeless and low-income children experienced significant adversity in their lives, with homeless preschool children facing more stress. However, differences in behavior as measured by the CBCL were minimal. Mothers' emotional status, in addition to various stressors, strongly predict children's negative outcomes for both CBCL subscales. These findings emphasize the importance of preventive family oriented interventions that address the needs of preschoolers and their mothers. PMID- 9200367 TI - Molecular genetics of childhood cancer: implications for pathogenesis, diagnosis, and treatment. PMID- 9200368 TI - Summary of major changes in the 1997 Red Book: Report of the Committee on Infectious Diseases. PMID- 9200369 TI - The adolescent varicocele: what's new with an old problem in young patients? PMID- 9200370 TI - Management of the young febrile child: a commentary on recent practice guidelines. PMID- 9200371 TI - Management of the young febrile child. Commentary on practice guidelines. PMID- 9200372 TI - Management of the young febrile child. Clinical guidelines in the setting of incomplete evidence. PMID- 9200373 TI - Management of the young febrile child: a continuing controversy. PMID- 9200374 TI - Plastic bronchitis: an unusual complication associated with sickle cell disease and the acute chest syndrome. PMID- 9200375 TI - Placental leptin: an important new growth factor in intrauterine and neonatal development? AB - BACKGROUND: Leptin, the protein product of the ob gene, is produced by the adipocyte and seems to function as a link between adiposity, satiety, and activity. Leptin has also been found to be necessary for pubertal development, conception, and pregnancy in mice, and is increased in prepubertal children, independent of adiposity, suggesting a role in childhood growth and development. This study investigated 100 mother/newborn pairs to determine the role of leptin in neonatal development. Placental tissue was assayed for leptin mRNA to evaluate it as a source of leptin production in utero. METHODS: One hundred mother/newborn pairs were enrolled in this study. Radioimmunoassay was performed for leptin on maternal venous and newborn cord blood. Leptin concentrations were measured in 43 children in Tanner stages 1 and 2 as a control group. Placental tissue was obtained from five mothers and assayed for leptin mRNA by reverse transcription/polymerase chain reaction (RT/PCR). Human placental cell lines JAR and JEG-3 were also assayed for leptin mRNA expression. RESULTS: Leptin was present in all newborns studied at a mean concentration of 8.8 ng/mL (+/-9.6 standard deviations). Leptin concentrations in cord blood correlated with newborn weight (r = .51), body mass index (BMI) (r = .48), and arm fat (r = .42). There was no correlation between leptin and insulin. When statistically covarying for adiposity for newborns and Tanner stages 1 and 2 children, newborns had greater concentrations of leptin (mean, 10.57 ng/mL) than children (mean, 3.04 ng/mL). Leptin was present in all mothers at a mean value of 28.8 ng/mL (+/-22.2 standard deviations). Leptin concentration correlated with prepregnancy BMI (r = .56), BMI at time of delivery (r = .74), and arm fat (r = .73). Maternal leptin correlated with serum insulin (r = .49). There was no correlation between maternal and newborn leptin concentrations. Thirteen percent of newborns had higher leptin concentrations than their mothers. Placental tissue from five separate placentas expressed leptin mRNA at comparable or greater levels than adipose tissue. Two human trophoblastic placental cell lines, JAR and JEG-3, also expressed leptin mRNA. CONCLUSIONS: The correlation between leptin and adiposity found in children and adults was also found in newborns. Serum leptin concentrations in newborns were increased more than three-fold compared with children in Tanner stages 1 and 2 when controlling for adiposity, suggesting that leptin concentrations in the newborn are not explained by adiposity alone. Maternal leptin concentrations correlated with measures of adiposity at delivery but did not correlate with newborn adiposity or leptin. Leptin mRNA was expressed both in placental tissue and in two human placental cell lines. These data suggest that leptin has a role in intrauterine and neonatal development and that the placenta provides a source of leptin for the growing fetus. PMID- 9200376 TI - Surgical treatment of craniosynostosis: outcome analysis of 250 consecutive patients. AB - OBJECTIVE: Surgery for craniosynostosis has evolved rapidly over the past two decades, with increased emphasis on early, extensive operations. Older published series may not accurately reflect more recent experience. Our study was designed to analyze outcome in a large series of consecutive patients treated recently at a single center. METHODS: We reviewed 250 consecutive patients who underwent surgical treatment of craniosynostosis between January 1, 1987 and December 31, 1992. They were divided into nine groups by suture involvement: sagittal, unilateral coronal, bilateral coronal, unilateral lambdoid, bilateral lambdoid, metopic, multiple suture, the Klee-blattschadel deformity (cloverleaf skull), and acquired craniosynostosis. Outcome was analyzed in terms of residual deformities and irregularities, complications, mortality, as well as the need for additional surgery. RESULTS: There were 157 males (62. 8%) and 93 females (37.2%), with most of the male preponderance accounted for by the large sagittal synostosis group, which consisted of 82 males and 25 females. Median age at first operation was 147 days. A named syndrome was present in 23 patients (9.2%) and was more common than expected with bilateral and unilateral coronal synostosis, the Kleeblattschadel deformity, and multiple suture synostosis. There were two deaths (0.8%), both with Klee-blattschadel patients, and 17 other complications (6.8%). Morbidity and mortality were significantly associated with secondary vs primary operations and syndromic vs nonsyndromic patients. Outcome analysis revealed the best surgical results with metopic synostosis and significantly less good results with the Kleeblattschadel deformity, multiple suture synostosis, and bilateral coronal synostosis. CONCLUSIONS: Using modern surgical techniques, craniosynostosis can be corrected with good outcomes and relatively low morbidity and mortality, particularly for otherwise healthy, nonsyndromic infants. PMID- 9200377 TI - Omeprazole-based dual and triple regimens for Helicobacter pylori eradication in children. AB - OBJECTIVE: To evaluate the efficacy and safety of omeprazole-based dual and triple regimens for the treatment of children with Helicobacter pylori infection. METHODS: Twenty-two patients (3 with gastric ulcer, 12 with duodenal ulcer, and 7 with nodular gastritis alone) were studied. Twelve ulcer patients also had nodular gastritis. The dual regimen included a 2-week course of omeprazole (0.6 mg/kg twice a day) and amoxicillin (30 mg/kg twice a day) (n = 10), and the triple regimen included the dual regimen plus clarithromycin (15 mg/kg twice a day) (n = 12). In patients with active ulcers, omeprazole once daily was administered for another 4 weeks. Endoscopic biopsies were taken before therapy and 4 weeks after completion of a 2-week course of therapy, and patients were followed for 6 months. The gastritis score (grade 0 to 3) and serum anti-H pylori IgG antibody titers were also determined. RESULTS: The regimens were tolerated by all patients. Eradication rates for the dual and triple regimens were 70% and 92%, respectively. Active ulcers completely healed within 6 weeks. Patients with nodular gastritis alone showed different clinical responses to therapy. Pretreatment histology showed chronic gastritis in all patients. Successful H pylori eradication significantly reduced the mean gastritis score from 2.9 to 1.3, but unsuccessful eradication did not reduce it. The disappearance of antral nodularity often coincided with the success of eradication. Successful eradication significantly decreased pretreatment serum anti-H pylori IgG antibody titers by 29% at 1 month, by 52% at 3 months, and by 67% at 6 months. Side effects were mild and were reported in 23% of patients. CONCLUSION: An omeprazole based regimen is safe and may be a better option for eradication of H pylori in children. Antral nodularity is a macroscopic marker of H pylori infection. PMID- 9200378 TI - Mortality, severe respiratory distress syndrome, and chronic lung disease of the newborn are reduced more after prophylactic than after therapeutic administration of the surfactant Curosurf. AB - OBJECTIVE: To test the hypothesis that prophylactic treatment with the surfactant Curosurf (Chiesi Farmaceutici SPA, Parma, Italy) improves survival and respiratory problems more than rescue treatment. DESIGN: Meta-analysis of three prophylaxis versus rescue treatment trials, conducted in four countries. METHODS: A meta-analysis was performed with the original, individual data of mortality, severe respiratory distress syndrome, and chronic lung disease of 671 newborns as outcomes. The random-effects logistic model (accounting for the trial-within country structure) was applied and adjusted for imbalances in covariates. RESULTS: The probability of each outcome differed between the countries, but the actual treatment effect itself did not. The adjusted odds ratios (ORs) and confidence intervals (CIs) for prophylaxis versus rescue were as follows: mortality: OR, .47; 95% CI, .30 to .73; severe RDS: OR, .50; 95% CI, .33 to .74; and chronic lung disease of the newborn in the survivors at day 28 after birth: OR, .54; 95% CI, .34 to .86. Gender, birth weight, gestational age, and prenatal administration of glucocorticosteroids were significant confounding covariates. CONCLUSION: The analysis shows that for the porcine surfactant Curosurf, prophylactic administration of surfactant has significant advantages over rescue therapy. PMID- 9200379 TI - Neurodevelopmental outcomes of Ugandan infants with human immunodeficiency virus type 1 infection. AB - BACKGROUND: The neurodevelopmental outcomes of human immunodeficiency virus type 1 (HIV-1)-infected Ugandan infants of nondrug-using mothers were studied using controlled, prospective methodology. METHOD: The sample of 436 full-term infants included 79 HIV-infected infants of HIV-1-infected mothers, 241 uninfected infants of HIV-1-infected mothers (seroreverters), and 116 uninfected infants born to HIV-negative mothers. Neurologic status, information processing ability, and motor and mental development were assessed from 6 to 24 months of age. Observations of caretaker-child interaction and home environments were made at 6 and 12 months. All evaluators were blinded to the HIV status of the child and family. RESULTS: Compared with seroreverters and uninfected infants, HIV-infected infants demonstrated greater deficits in motor development and neurologic status, and more frequent and earlier onset of motor and neurologic abnormalities. Compared with controls, HIV-infected infants had more abnormalities in mental development at 6 and 18 months and an earlier onset of abnormalities. By 12 months, 30% of HIV-infected infants demonstrated motor abnormalities and 26% cognitive abnormalities as compared with 11% and 6% among seroreverters and 5% and 6% among seronegative infants. HIV-infected infants (62%) demonstrated a higher probability of developing an abnormal neurologic examination by 12 months, compared with seroreverters (17%) or seronegative infants (15%). Information processing abilities did not differ as a function of HIV infection. Home environments and infants' interactions with caretakers were similar across groups. CONCLUSION: We conclude that HIV infection results in more frequent and earlier abnormalities in infants' neurologic status and motor development that are not attributable to other biological and environmental risk factors. More frequent mental developmental abnormalities were evident at several ages. However, information-processing abilities, such as recognition memory, may be spared from HIV-related deficits. PMID- 9200380 TI - Human granulocyte colony-stimulating factor may improve outcome attributable to neonatal sepsis complicated by neutropenia. AB - OBJECTIVES: To determine whether adjunctive therapy with recombinant human granulocyte colony-stimulating factor (rhG-CSF) could reverse sepsis-associated neonatal neutropenia and improve neonatal survival compared with conventional therapy in a phase I/II-type trial. STUDY DESIGN: An intravenous infusion of rhG CSF (10 microg/kg/d x 3 d) was administered to 14 septic neutropenic neonates. Neutrophilic responses and outcome of these neonates were compared with 11 concurrently treated, retrospectively selected, case-matched control septic patients identified by using a search of medical records coded for sepsis with neutropenia (>/=24 hours). RESULTS: Seven neonates with early-onset sepsis with neutropenia at birth and seven neonates with late-onset sepsis plus neutropenia (all with necrotizing enterocolitis) were entered in the rhG-CSF treatment group. Results were compared with a conventional therapy control group (five early onset, six late onset). No significant differences existed in the birth weight, gestational age, use of antibiotic therapy, magnitude of respiratory support, severity of metabolic acidosis, use of vasopressors, or other supportive therapy between the two groups. In the rhG-CSF-treated group and in the conventionally treated control group, the absolute neutrophil count (ANC) (mean +/- SEM) was 585 +/- 138 and 438 +/- 152, respectively. The ANC increased to more than baseline in the rhG-CSF-treated group by 10-fold versus 2-fold at 24 hours, 18-fold versus 4 fold at 48 hours, 24-fold versus 5-fold at 72 hours (significant by one-way analysis of variance in the rhG-CSF group only), and 29-fold versus 16-fold at 7 to 10 days when compared with the conventional therapy group. There were no nonresponders in the rhG-CSF group by 24 hours after the first dose of study drug. Monocyte cell counts also increased significantly in both groups by 7 days after entry into this protocol but remained within normal range for age. No clinically significant effect on lymphocytes, erythrocytes, or platelet counts was noted. Thirteen patients in the rhG-CSF-treated group (92%; 13 out of 14) and five in the conventionally treated group (55%; 5 out of 11) survived to 28 days after the onset of the signs of sepsis. No adverse effects were noted in the rhG CSF-treated group. CONCLUSIONS: rhG-CSF can increase the neutrophil count in critically ill septic neutropenic neonates. This finding suggests that rhG-CSF may be effective in a therapeutically useful time frame to treat septic neonates with neonatal neutropenia attributable to bone marrow suppression or neutrophil consumption. Future randomized trials are needed to validate the beneficial effects of rhG-CSF and to determine whether any significant side effects of therapy exist. PMID- 9200381 TI - Growth failure as a prognostic indicator of mortality in pediatric HIV infection. AB - OBJECTIVE: To study the effect of perinatally acquired human immunodeficiency virus (HIV) on somatic growth and examine the relationship of nutritional status to mortality in HIV-infected infants. METHOD: Pregnant women attending the antenatal clinic at Mulago hospital in Kampala, Uganda, were enrolled. All live born babies born to HIV-1 seropositive (HIV+) women, and to every fourth age matched HIV-1 seronegative (HIV-) woman, were followed for 25 months. RESULTS: The mean weight-for-age and length-for-age curves of HIV+ children were significantly lower than those of HIV- controls and seroeverters. Forty-five (54%) of the 84 HIV+ infants died before their second birthday, as compared with a 1.6% and 5.6% mortality in HIV- and seroeverters. HIV+ infants with an average weight-for-age Z-score below -1.5 in the first year of life have a nearly fivefold risk of dying before 25 months of age compared with noninfected controls. CONCLUSION: Perinatally acquired HIV infection is associated with early and progressive growth failure. The severity of growth failure is associated with an increased risk of mortality. The effect of early, aggressive nutritional intervention in delaying HIV progression and mortality should be evaluated by controlled intervention studies. PMID- 9200382 TI - School-related issues among HIV-infected children. AB - OBJECTIVE: Many children with human immunodeficiency virus (HIV) infection are surviving long enough to reach school age. This study describes issues related to school attendance and disclosure of HIV infection in a population of HIV-infected children. METHODS: A statewide pediatric HIV surveillance system was used to collect data on school-age (>/=5 years old) HIV-infected children. In addition, HIV clinic nurses familiar with the child's history participated in a cross sectional survey that collected information on school-related issues during the 1993-1994 school year. RESULTS: Of the 92 school-age children, only 3 were too ill to attend school. Another 5 children were home-schooled. Of the 84 who attended school outside the home, 25% had severe symptoms of HIV infection (Centers for Disease Control and Prevention [CDC] clinical category C). Absence from school ranged from less than 2 weeks during the year for half of the children (51%) to more than 8 weeks for 9 children (12%). Twenty-nine percent of the children received medication in school, usually administered by the school nurse. Over two thirds of the 50 children ages 5 to 10 years had not been told that they had HIV infection. Only 1 of the 20 children more than 10 years of age was not aware of her HIV infection. For 53% of the children attending school, no school personnel had been informed of the child's HIV infection. Administration of HIV medications at school, age of child, and treatment at one particular HIV clinic were associated with the parents' decision to inform school personnel. In the 47% of cases where the school had been informed, school nurses were most frequently notified, followed by principals and teachers. CONCLUSION: Only 3% of school-age children were too ill to attend school, and almost all were enrolled in public schools. The number of HIV-infected children reaching school age will continue to grow, and public schools will bear the responsibility for educating these children. Health care providers will increasingly be called upon for guidance by both educators and families to assure that HIV-infected children receive the best education possible. PMID- 9200383 TI - Congenital lymphocytic choriomeningitis virus syndrome: a disease that mimics congenital toxoplasmosis or Cytomegalovirus infection. AB - OBJECTIVE: To describe the clinical characteristics of intrauterine infection with lymphocytic choriomeningitis (LCM) virus, an uncommonly recognized cause of congenital viral infection. PATIENTS: Three infants born in the midwestern United States in 1994 and 1995 with clinical features and serologic studies consistent with congenital LCM virus infection and cases of congenital infection identified by review of the medical literature between 1955 and 1996. RESULTS: Twenty-six infants with serologically confirmed congenital LCM virus infection were identified. Twenty-two infants were products of term gestations, and birth weights ranged from 2384 to 4400 g (median, 3520 g). Ocular abnormalities, macrocephaly, or microcephaly were the most commonly identified neonatal features. Twenty-one infants (88%) had chorioretinopathy, 10 (43%) had macrocephaly (head circumference >90th percentile) at birth, and 3 (13%) were microcephalic (head circumference <10th percentile). Macrocephaly and hydrocephalus developed postnatally in one of the latter infants. Hydrocephalus or intracranial calcifications were documented in five infants by computed tomography or magnetic resonance imaging. Nine infants (35%) died, and 10 (63%) of the 16 reported survivors had severe neurologic sequelae, consisting of spastic quadriparesis, seizures, visual loss, or mental retardation. One-half of the mothers reported illnesses compatible with LCM virus infection, and 25% reported exposures to rodents during their pregnancies. CONCLUSIONS: These cases suggest that congenital LCM virus infection could be an underrecognized cause of congenital infection among infants born in the United States. Because of the clinical similarities of these congenital infections, cases of congenital LCM virus infection can be confused with infections with cytomegalovirus or Toxoplasma gondii. PMID- 9200384 TI - Human monocytic ehrlichiosis in children. AB - BACKGROUND: Much of what is known about human monocytic ehrlichiosis (HME) is based upon studies with adult patients. PURPOSE: To review our experience with HME to better understand the epidemiology, clinical manifestations, and outcome of this disease in children. METHODS: Demographic, clinical, and laboratory data were gathered after review of the medical records of patients identified with HME. RESULTS: Twelve patients with an median age of 7.4 years (range, 7 months to 13.7 years) were identified with HME; 10 were white, 7 were male, and 10 were from hometowns of <800 people. Eight patients presented from May through July, and 8 had a history of tick bites. Symptoms demonstrated by the patients during their illness included fever (100%), rash (67%), myalgias (58%), and vomiting, diarrhea, and headache (25%). On presentation, patients demonstrated thrombocytopenia (92%), elevated liver function tests (91%), lymphopenia (75%), hyponatremia (67%), leukopenia (58%), and anemia (42%) on the initial laboratory examination. Four patients presented in shock and 3 required blood pressure support and mechanical ventilation for a median of 10 days (8 to 37 days). These complicated patients required longer hospitalization (19.5 days vs 5. 5 days) and attained higher blood urea nitrogen levels (42.5 mg/dL vs 10 mg/dL) than the patients not presenting with shock. Morbidity associated with HME patients included a decrease in cognitive and neurologic performance. CONCLUSIONS: More information and long-term follow-up is required to understand the full spectrum of disease and morbidity associated with HME in children. PMID- 9200385 TI - Does the supine sleeping position have any adverse effects on the child? I. Health in the first six months. The ALSPAC Study Team. AB - OBJECTIVE: To assess whether the recommendations that infants sleep supine could have adverse health consequences. DESIGN: A prospective study of infants, delivered before, during, and after the Back to Sleep Campaign in the United Kingdom (UK), followed to 6 months of age. The children were part of the Avon Longitudinal Study of Pregnancy and Childhood (ALSPAC). Subjects. Singletons born to mothers resident in the three former Bristol-based health districts of Avon in the period June 1991 to December 1992, and for whom questionnaires were completed on sleeping position at 4 to 6 weeks of age (n = 9777); for these infants 8524 questionnaires were also completed at 6 to 8 months of age. MAIN OUTCOME MEASURES: Subjective measures of health, the presence of specific signs and symptoms, duration of sleep at night, and calling the family doctor to the home. RESULTS: Of 43 outcomes considered, after adjustment for 12 factors using logistic regression only 2 were associated with raised risk among infants put to sleep on their back (diaper rash and cradle cap). Infants put to sleep prone had increased risk of a number of health outcomes, including cough and possibly pyrexia. CONCLUSIONS: There is no evidence that putting infants to sleep in the supine position results in increased morbidity, although changes in prevalence of rare disorders would not have been identified. PMID- 9200386 TI - Predictors of hemolytic uremic syndrome in children during a large outbreak of Escherichia coli O157:H7 infections. AB - OBJECTIVE: To evaluate risk factors for progression of Escherichia coli O157:H7 infection to the hemolytic uremic syndrome (HUS). STUDY DESIGN: We conducted a retrospective cohort study among 278 Washington State children <16 years old who developed symptomatic culture-confirmed E coli O157:H7 infection during a large 1993 outbreak. The purpose of the study was to determine the relative risk (RR) of developing HUS according to demographic characteristics, symptoms, laboratory test results, and medication use in the first 3 days of illness. RESULTS: Thirty seven (14%) children developed HUS. In univariate analysis, no associations were observed between HUS risk and any demographic characteristic, the presence of bloody diarrhea or of fever, or medication use. In multivariate analysis, HUS risk was associated with, in the first 3 days of illness, use of antimotility agents (odds ratio [OR] = 2.9; 95% confidence interval [CI] 1.2-7.5) and, among children <5.5 years old, vomiting (OR = 4. 2; 95% CI 1.4-12.7). Among the 128 children tested, those whose white blood cell (WBC) count was >/=13 000/microL in the first 3 days of illness had a 7-fold increased risk of developing HUS (RR 7. 2; 95% CI 2.8-18.5). Thirteen (38%) of the 34 patients with a WBC count >/=13 000/microL developed HUS, but only 5 (5%) of the 94 children whose initial WBC count was <13 000/microL progressed to HUS. Among children who did not develop HUS, use of antimotility agents in the first 3 days of illness was associated with longer duration of bloody diarrhea. CONCLUSIONS: Prospective studies are needed to further evaluate measures to prevent the progression of E coli O157:H7 infection to HUS and to assess further clinical and laboratory risk factors. These data argue against the use of antimotility agents in acute childhood diarrhea. Our finding that no intervention decreased HUS risk underscores the importance of preventing E coli O157:H7 infections. PMID- 9200387 TI - Sequential external counterpulsation increases cerebral and renal blood flow. AB - The purpose of this study was to evaluate the effect of sequential external counterpulsation (SECP) on cerebral and renal blood flow. The effect of SECP on carotid and renal artery blood flow was studied in 35 and 18 patients, respectively. With a portable unit, cuffs were applied to the calves and thighs, sequentially inflated with air at the onset of diastole, and deflated at the onset of systole. Carotid and renal artery Duplex studies were performed during intermittent SECP. Flow velocity and flow velocity integral were measured at baseline and during SECP. Diastolic augmentation of carotid and renal artery flow velocity was observed in all patients. The mean carotid flow velocity integral increased by 22% from 27.7 +/- 1.8 cm to 33.1 +/- 2.3 cm (P = 0.001). The mean renal artery flow velocity integral increased by 19% from 21 +/- 1 cm to 25 +/- 1 cm (P = 0.0001). With SECP, a new diastolic Doppler flow velocity wave was observed, with an average peak carotid diastolic flow velocity of 56 +/- 4 cm/sec and an average peak renal artery diastolic flow velocity of 40 +/- 2.5 cm/sec. This diastolic wave was 75% (carotid) and 68% (renal) as high as the systolic wave during SECP. In addition, with SECP the systolic wave increased by 6% and 8% in the carotid and renal artery, respectively (P = 0.02 and 0.006, respectively). In conclusion, SECP significantly increases carotid and renal blood flow. This noninvasive, harmless treatment may be useful to support patients with decreased cerebral and renal perfusion. PMID- 9200388 TI - Salutary effect of adjunctive intracoronary nicorandil administration on restoration of myocardial blood flow and functional improvement in patients with acute myocardial infarction. AB - Salutary effect of nicorandil, a K+ adenosine triphosphate channel opener, on restoration of myocardial blood flow and functional improvement after coronary revascularization was investigated in 20 patients with first anterior acute myocardial infarction. Ten patients received intracoronary administration of nicorandil (2 mg) after coronary revascularization; the other 10 patients received coronary revascularization only and served as control subjects. Myocardial contrast echocardiography and two-dimensional echocardiography were performed to assess microvascular integrity and regional function in the infarcted area. Nicorandil improved peak contrast intensity ratio (p < 0.001), calculated as the ratio of peak contrast intensity in the infarcted and noninfarcted areas, indicating the restoration of myocardial blood flow to the infarcted myocardium. Regional wall motion improved more significantly in 1 month in patients who received nicorandil (p < 0.01). Thus our results suggested the usefulness of intracoronary nicorandil administration after coronary revascularization for restoring blood flow and functional improvement in patients with acute myocardial infarction. PMID- 9200389 TI - Glucose tolerance status and severity of coronary artery disease in men referred to coronary arteriography. AB - Increasing attention is being paid to disturbances in glucose metabolism as key explanatory factors for the development of coronary artery disease. We studied the prevalence of impaired glucose tolerance and non-insulin-dependent diabetes and the levels of plasma insulin after an oral glucose tolerance test in 99 men with heart disease but without a history of diabetes referred to coronary arteriography; we also compared the outcome with a matched control group (n = 116). The severity of atherosclerosis in coronary angiograms was evaluated according to glucose tolerance status. Among the 99 patients with coronary artery disease, 37.4% had an abnormal oral glucose tolerance test result, whereas only 18.1% of the control group had an abnormal result (p < 0.01). Moreover, patients with heart disease and normal glucose tolerance were hyperinsulinemic compared with the control group (p < 0.01). By analysis of variance no statistically significant difference in severity of coronary atherosclerosis on coronary angiograms was found. In conclusion, we demonstrated frequent disturbances in glucose metabolism indicating insulin resistance in patients with ischemic heart disease without a history of diabetes, but we could not demonstrate a relation between these disturbances and degree of coronary atherosclerosis. PMID- 9200390 TI - Selection of thrombolytic therapy for individual patients: development of a clinical model. GUSTO-I Investigators. AB - We developed a logistic regression model with data from the GUSTO-I trial to predict mortality rate differences in individual patients who received accelerated tissue plasminogen activator (TPA) versus streptokinase treatment for acute myocardial infarction. A nomogram was developed from a reduced version of this model that approximated the underlying risk of patients treated with streptokinase, and thus the benefit of TPA. The 30-day mortality rate with accelerated TPA was 0.063 versus 0.073 with streptokinase and subcutaneously administered heparin and 0.074 with streptokinase and intravenously administered heparin. No baseline patient characteristics were significantly associated with a different relative effect of TPA. Older patients and those with anterior infarction, higher Killip classification (except Killip class IV), lower blood pressure, and increased heart rate had the greatest absolute benefit with accelerated TPA. Patients with acute myocardial infarction who had more high-risk characteristics derived a greater absolute benefit from treatment with accelerated TPA versus streptokinase. PMID- 9200391 TI - Quantitative measures of regional asynergy add independent prognostic information to left ventricular ejection fraction in patients with prior myocardial infarction. AB - The purpose of this study was to determine if quantitative measurements of regional asynergy add independent prognostic information to global ejection fraction in patients with chronic coronary artery disease. Four hundred eighty six patients with a history of Q-wave myocardial infarction who underwent gated equilibrium radionuclide angiography at least 3 months after infarction were monitored for a median duration of 4.7 years. During follow-up there were 95 deaths. Four of five regional asynergy indexes analyzed were associated with overall mortality. The strength of the association between overall mortality and the index that proved to be optimal (univariate chi2 = 26.4, p < 0.001) was stronger than for global ejection fraction (univariate chi2 = 21.5, p < 0.001). For patients with global ejection fraction <40%, 4-year survival was 87% for those with a low asynergy index versus 65% for those with a high asynergy index (p = 0.016). In conclusion, indexes of regional asynergy add independent prognostic information to global left ventricular ejection fraction. PMID- 9200392 TI - Treatment of medically and surgically refractory angina pectoris with high thoracic epidural analgesia: initial clinical experience. AB - Surgical sympathectomy can relieve symptoms of angina in patients with refractory angina. However, in these high-risk patients this thoracic surgery may result in significant morbidity and mortality rates. Similar sympathetic blockade can now be produced with high thoracic epidural analgesia (HTEA). From September 1995 to August 1996, we treated 10 consecutive patients with HTEA. These eight men and two women, aged 58 +/- 5 years, with extensive three-vessel coronary disease and ejection fractions of 40% +/- 5%, had New York Heart Association (NYHA) class IV angina despite medical therapy, including nitrates, beta-blockade, calcium channel blockade, and narcotics. HTEA was performed at the T1 through T4 levels with a catheter placed either percutaneously or surgically, with radiographic confirmation of catheter placement with an epidurogram or computed tomography scan. Bupivacaine (0.25% to 0.5%), an amide local anesthetic, was given as a bolus through the epidural catheter and then maintained either as a continuous infusion or an intermittent rebolus. The epidural catheter remained in place for 7 days in four patients, 14 days in three patients, and > or =90 days in three patients. Before consideration for HTEA, each patient was deemed unsuitable for or refused coronary bypass surgery and percutaneous coronary angioplasty and had NYHA class IV symptoms of angina. Seven of 10 patients required intravenous nitroglycerin and heparin and were unable to be discharged from the intensive care unit because of anginal symptoms. Two of these seven patients also required an intraaortic balloon pump for symptom control. After HTEA, all 10 patients had improved symptoms, with five patients improving to NYHA class II symptoms and five improving to NYHA class III. All seven patients receiving intravenous nitroglycerin, heparin, or intraaortic balloon pump support had these modalities discontinued. Six of these seven patients were subsequently discharged from the hospital. One patient died from a non-HTEA related cause. There were no HTEA related deaths. There were three catheter-related complications necessitating catheter removal during 12 months of HTEA use. Local infection developed in one patient, one had catheter occlusion caused by fibrosis, and one patient had chronic back pain exacerbation from a paraspinous muscle spasm. No patient had a myocardial infarction or a significant arrhythmia. In patients with otherwise intractable angina pectoris, HTEA is an effective modality that produces symptomatic relief of angina pectoris and allows increased activity level. PMID- 9200393 TI - Long-term implications of racial differences in the use of revascularization procedures (the Myocardial Infarction Triage and Intervention registry). AB - The purpose of this study was to determine if less intensive use of revascularization procedures in black patients influenced the long-term survival of patients hospitalized for acute myocardial infarction (AMI) in metropolitan Seattle. From January 1988 through June 1994, AMI developed in 420 (4%) black and 10,834 white patients before hospital discharge or death. Black patients were 6 years younger, more socioeconomically disadvantaged, and had more hypertension, diabetes, and less prior coronary surgery. Similar proportions of black and white patients received thrombolytic therapy or cardiac catheterization. However, during hospitalization, 18% of black patients underwent coronary angioplasty compared with 26% of white patients (p = 0.0004); coronary artery bypass surgery was also used less frequently in black patients (7% vs 12%, p = 0.002). Unadjusted 2-year survival was 79% for black patients and 77% for white patients (p = 0.12). After adjusting for age, clinical variables, and the use of cardiac catheterization, thrombolytic therapy, and revascularization, race was not associated with long-term survival (hazard ratio = 0.92, 95% confidence interval = 0.73 to 1.17). Despite the less intensive use of revascularization procedures in black patients in Seattle, long-term survival after AMI was similar in the two groups of patients. PMID- 9200394 TI - Clinical characteristics and long-term outcome of patients in whom congestive heart failure develops after thrombolytic therapy for acute myocardial infarction: development of a predictive model. AB - Ischemic heart disease is the most common cause of congestive heart failure, which often begins after acute myocardial infarction. To better delineate the clinical characteristics and outcomes of patients in whom congestive heart failure develops after acute myocardial infarction in the thrombolytic era, we prospectively evaluated patients enrolled in six of the TAMI trials. The study cohort comprised 1619 consecutive patients who had at least 1 mm of ST-segment elevation in two contiguous electrocardiographic leads within 6 hours of the onset of acute myocardial infarction and who received intravenous thrombolytic therapy. We prospectively collected clinical characteristics, baseline demographics, acute and 1-week angiographic variables, and in-hospital and 1-year outcome data. We performed stepwise multivariable regression analysis to determine the noninvasive and invasive predictors of the development of in hospital congestive heart failure. Congestive heart failure developed in 301 patients in the hospital (19% of 1521 patients admitted were not in heart failure). These patients were likely to be older and female, have diabetes mellitus and previous myocardial infarction, and have an anterior wall myocardial infarction. On acute angiography, they had lower ejection fractions and a higher incidence of multivessel disease. Patency at 90 minutes was lower in the patients with congestive heart failure, and acute mitral regurgitation occurred in 1.6% versus 0.21% of patients without congestive heart failure. Patients with congestive heart failure had higher mortality, more in-hospital complications, and longer hospitalizations. At 1-year follow up, 21% of the patients in whom congestive heart failure developed had died versus 5% in the group without congestive heart failure. Predictors of new congestive heart failure included increased age, anterior wall myocardial infarction, lower pulse pressure and systolic blood pressure, diabetes mellitus, and the presence of rales on admission. The acute angiographic variables of reduced ejection fraction, increased number of diseased vessels, and attempted percutaneous intervention improved the concordance of the predictive model by 6%. Congestive heart failure remains a common clinical problem after acute myocardial infarction and is associated with a twofold increase in in-hospital morbidity and a fourfold increase in in-hospital and 1-year mortality. The development of congestive heart failure in the hospital can be predicted from noninvasive and invasive baseline characteristics. We present a simple table to predict congestive heart failure from baseline characteristics and invasive information. PMID- 9200395 TI - Ventricular pacing with a novel gastroesophageal electrode: a comparison with external pacing. AB - Temporary endocardial pacing is a technically demanding invasive procedure requiring sterile precautions and access to fluoroscopy. External (transcutaneous) pacing requires high current for capture and is poorly tolerated in the conscious patient. An esothoracic pacing system has been developed capable of reliable ventricular capture. The flexible gastroesophageal electrode is passed into the stomach. The distal 6 cm is angled to 90 degrees with an internal pulley system, positioning the tip of the gastroesophageal electrode in the fundus of the stomach. Ventricular pacing is performed with a spherical electrode (cathode) mounted on the gastroesophageal electrode tip in conjunction with a chest pad (anode) positioned medial to the cardiac apex. Of 91 subjects in which esothoracic pacing was attempted, 86 (94.5%) demonstrated successful ventricular capture at the maximum pulse duration used (40 msec). Threshold current for ventricular capture ranged from 22.5 +/- 8.1 mA at a pulse duration of 40 msec to 29.9 +/- 8.6 mA at a pulse duration of 10 msec. Esothoracic pacing was compared with external pacing in a subgroup (n = 30) of patients. Ventricular capture with the gastroesophageal electrode was more common when compared with the external approach (27 [90%] of 30 vs 13 [43.3%] of 30, p < 0.001). In those subjects in whom ventricular capture was obtained with both methods, threshold current for capture was significantly lower with the esothoracic approach. This gastroesophageal electrode may be useful in the emergency management of acute bradyarrhythmias. PMID- 9200396 TI - Automated morphometry of coronary arteries with digital image analysis of intravascular ultrasound. AB - We designed and tested digital image processing strategies to perform fully automated segmentation of luminal and medial-adventitial boundaries in intravascular ultrasound images of human coronary arteries. Automated segmentation is an essential tool for advanced techniques of clinical visualization and quantitative measurement. Vascular compliance measurements and three-dimensional reconstructions are demonstrated as examples of such applications. Digital image processing was performed in three phases: (1) preprocessing, including a polar transform, local contrast enhancement, and speckle noise filtering; (2) segmentation, involving radial scanning, region growing, or cost-function minimization techniques; and (3) postprocessing, involving dropout filtering and outline smoothing. Cross-sectional areas were compared with manual tracings from experienced operators and showed good agreement. The algorithm bias ranged from -0.34 to 1.18 mm2; interclass and intraclass correlation coefficients ranged from 0.83 to 0.94. The designed techniques currently allow fully automated segmentation without operator interaction of the luminal and, if present, medial-adventitial boundary. PMID- 9200397 TI - Increased left ventricular cavity size, not wall thickness, potentiates myocardial ischemia. AB - Left ventricular (LV) hypertrophy increases the vulnerability of the myocardium to ischemia. The purpose of this study was to determine whether LV diameter or wall thickness was the principal determinant of the effect of LV mass on the development of ischemia, measured by exercise thallium perfusion imaging, in a population with coronary artery disease (CAD). We studied 109 patients with CAD but no prior myocardial infarction who underwent exercise thallium imaging within 1 year of coronary angiography. Thallium perfusion defects were present in 76% of patients. LV mass index was associated with thallium perfusion abnormalities (odds ratio 2.09 for 50 gm increments), an association that persisted after adjusting for extent of CAD. LV end-diastolic diameter had a strong correlation with a thallium defect (odds ratio 3.7 for 10 mm increments), but LV wall thickness had no correlation (odds ratio 1.0 for 5 mm increments). In a stepwise regression model that included extent of CAD and other potential clinical variables, LV end-diastolic diameter was the strongest predictor of thallium defects (adjusted odds ratio 4.5). This study confirms the association of LV hypertrophy with ischemia in patients with CAD, specifically in patients with eccentric hypertrophy. PMID- 9200398 TI - Allograft aortopathy: an in vivo study of donor aorta involvement in cardiac allograft vasculopathy. AB - Limited histopathologic studies of failed cardiac allografts have demonstrated that cardiac allograft vasculopathy extends into the donor aorta; however, no study has examined the development of allograft aortic intimal proliferation in vivo in conjunction with coronary intimal hyperplasia. By using simultaneous intracoronary and intraaortic ultrasound, we studied 20 consecutive heart transplant recipients at 2.5 +/- 2.1 years after transplantation. The degree of coronary intimal thickening was strongly correlated with the development of intraaortic intimal hyperplasia (r = 0.90; p < 0.0001). Multivariate predictors of aortic intimal thickening included years after transplant (r = 0.47; p = 0.03), serum cholesterol level (r = 0.65, p = 0.003), and serum triglyceride level (r = 0.51; p = 0.03). Allograft aortopathy occurs in a similar manner to allograft coronary disease, thus providing support for the notion that an immunologic stimulus operating across the allograft vascular bed may be responsible for the development of cardiac allograft vasculopathy. Furthermore, this investigation provides insight into the putative role of hyperlipidemia in allograft vascular disease. PMID- 9200399 TI - Evolving trends in the epidemiologic factors of heart failure: rationale for preventive strategies and comprehensive disease management. PMID- 9200400 TI - Diastolic myocardial velocity measurements. PMID- 9200402 TI - Growth retardation in children with chronic renal insufficiency. PMID- 9200401 TI - Concomitant nitroglycerin and rTPA. PMID- 9200403 TI - Pathogenesis of acute renal failure: new aspects. AB - Based on 2 case presentations - acute renal failure (ARF) due to myeloma kidney and due to angiotensin-converting enzyme inhibitor administration in the presence of transplant artery stenosis - new aspects in the pathogenesis of ARF are presented and discussed. The multifactorial pathogenesis of ARF includes (a) a disturbance of glomerular microcirculation (afferent and perhaps mesangial constriction, inadequate efferent dilatation); (b) a disturbance of medullary microcirculation (medullary capillary congestion) attributed to a combination of endothelial damage and tubular dilatation; (c) tubular cell damage which, though rarely in humans justifying the term 'acute tubular necrosis', promotes both backleak of glomerular filtrate and shedding of brush border vesicles; (d) the latter promotes tubular obstruction by casts which consist of Tamm-Horsfall protein and brush border components. Once ARF is established, repair processes set in which appear to depend on growth factors such as epidermal growth factor and insulin-like growth factor 1, of which there is a relative shortage in established ARF. Experimental therapeutic approaches focus on the restitution of microcirculation (endothelin receptor antagonists, atriopeptins), interference with cast formation (integrin receptor blockers), and the promotion of recovery by growth factors. PMID- 9200404 TI - Glomerular volume in congestive cardiac failure. AB - Glomerular volume has been reported to be increased in patients with congenital cyanotic heart disease and cor pulmonale; however it has not been systematically studied in patients with congestive cardiac failure (CCF). Glomerular volume was therefore measured by point-counting serially sectioned glomerular profiles of 25 randomly selected glomeruli using the Cavalieri principle in autopsy specimens from 8 patients dying from CCF and 6 age-matched controls with no renal or cardiac pathology. Mean glomerular volume was not different between patients dying from CCF and controls, 2.49 (0.21) vs. 2.25 (0.26) x 10(6) microm3, and the distribution of individual glomerular volumes was similar in the two groups. We conclude that severe CCF is not associated with significant glomerular enlargement and that the previously reported glomerular enlargement in cyanotic heart disease is likely to be mediated through hypoxemia. PMID- 9200405 TI - Renal function in cancer patients treated with hyperthermic isolated limb perfusion with recombinant tumor necrosis factor-alpha and melphalan. AB - Hyperthermic isolated limb perfusion (HILP) with recombinant tumor necrosis factor-alpha (r-TNF alpha) and melphalan has been shown to result in a sepsis like syndrome due to leakage of r-TNF alpha from the perfusion system to the systemic circulation. We have studied renal function parameters in 11 cancer patients, who underwent 12 perfusions. Three patients, perfused with melphalan only, served as controls. All patients treated with r-TNF alpha developed a sepsis syndrome and needed volume replacement and inotropes to remain normotensive; controls had an uneventful postoperative course. Creatinine clearance decreased transiently on the day of perfusion in both groups (mean preperfusion clearance 118 ml/min, mean post-perfusion clearance 68 ml/min, p < 0.02, n = 15). Follow-up measurements of renal plasma flow and glomerular filtration rate in 9 r-TNF alpha-treated patients did not suggest permanent damage. One patient became hypotensive and developed transient multiple organ dysfunction with renal failure needing hemofiltration. In r-TNF alpha-treated patients, but not in controls, a transient increase in clearance of beta2 microglobulin (0.05 vs. 8 ml/min, p < 0.001) and urinary excretion of phosphate (12 vs. 48 mmol/l, p < 0.05) was seen, compatible with proximal tubular dysfunction. These data suggest that HILP with melphalan decreases glomerular function, whether or not r-TNF alpha is added to the perfusion circuit. Extension of the treatment regimen with r-TNF alpha may result in additional proximal tubular dysfunction. If hypotension can be avoided, this deterioration in renal function seems to be transient, with full recovery within weeks. PMID- 9200406 TI - Histological localization of advanced glycosylation end products in the progression of diabetic nephropathy. AB - We studied the immunohistochemical localization of advanced glycosylation end products (AGEs) in the progression of diabetic nephropathy. Fourteen NIDDM patients with diabetic nephropathy were evaluated: 2 patients with normoalbuminuria, 4 with microalbuminuria (MA) and 8 with overt proteinuria (OP). Three patients with minor glomerular abnormalities were used as nondiabetic controls. Immunoreactivity to a monoclonal anti-AGE antibody (6D12) was recognized on the internal elastic membranes of arterial walls in every diabetic group. Hyaline lesions of arterioles of the MA and OP groups demonstrated strong reactions with 6D12. A portion of the nodular and exudative lesions in glomeruli of OP group patients also revealed immunoreactivity to 6D12. No immunoreactivity to 6D12 was observed in nondiabetic control specimens. We confirm that the accumulation of AGEs began in arterial walls of the early stage and presented in glomerular lesions of the late stage of the progression of diabetic nephropathy. PMID- 9200407 TI - Increased expression of platelet-derived growth factor A and collagenous matrix proteins in congenital multicystic renal dysplasia. AB - The expression of platelet-derived growth factor A (PDGF-A), and its spatial and temporal relationship to interstitial collagens in kidneys with congenital multicystic dysplasia using in situ hybridization, have been examined. Seventeen dysplastic kidneys (16 weeks to 7 months) and 20 normal age-matched controls were used in the study. Increased PDGF-A mRNA was detected in dysplastic compared to normal kidneys in all age groups including extensively fibrotic postnatal kidneys. An abundant PDGF-A mRNA signal was seen within the epithelial cells of cystically dilated or dysplastic tubules and within interstitial fibroblasts and disorganized primitive mesenchyme. A comparable amount of PDGF-A protein was detected by Western blotting. Procollagen I and III mRNA were increased in fibroblasts surrounding cystic and dysplastic tubules. We conclude that tubular epithelial production of PDGF-A may induce collagenous matrix production by adjacent fibroblasts, while marked up-regulation of PDGF-A by interstitial cells may be responsible for sustainable fibrogenic effects in the fetal kidney contributing to renal maldevelopment. PMID- 9200408 TI - GB virus C and hepatitis C virus infections in hemodialysis patients in eight Japanese centers. AB - RNA of a putative non-A to E hepatitis virus, designated GB virus C (GBV-C), was detected in 40 (6.2%) of 645 hemodialysis patients, at a frequency significantly higher than in 3 (0.9%) of 336 blood donors in Japan (p < 0.001). A history of transfusion was more frequent (88 vs. 58%, p < 0.001), the duration of dialysis was longer (13.2 +/- 7.9 vs. 7.9 +/- 6.5 years, p < 0.001), and the detection of hepatitis C virus RNA was more often (38 vs. 18%, p < 0.01) in the 40 patients with GBV-C RNA than in the 605 patients without it. The prevalence of GBV-C RNA varied widely from 0 to 10% among the 8 dialysis centers. These results indicate that hemodialysis patients would be at increased risk of GBV-C transmitted by transfusions. The detection of GBV-C RNA in the 5 patients without a history of transfusion and a high prevalence restricted to certain dialysis centers would reflect nosocomial infection. PMID- 9200409 TI - Defect of cell-mediated immune response against hepatitis B virus: an indication for pathogenesis of hepatitis-B-virus-associated membranous nephropathy. AB - To elucidate the questions of why not all patients with hepatitis B virus (HBV) infection develop HBV membranous nephropathy (HBVMN), we first measured serum HBe circulating immune complex (CIC) during the acute nephrotic phase of HBVMN and in HBV carriers. We found that the level of HBe CIC was low in the HBVMN patients and absent either in HBsAg+/HBeAg+ patients without HBVMN or HBsAg+/HBeAg- asymptomatic carriers. Second, we needed to characterize the cellular immune response to HBV in patients with HBVMN. However, lack of a suitable autologous effector/target cell system makes a precise study of HBVMN pathogenesis difficult. In the present study, we established a model system by using autologous HBcAg-expressing Epstein-Barr-virus-immortalized lymphoblastoid cell lines (LCL) as stimulator/target cells. Both proliferative response after stimulation with HBcAg and cytotoxic activity against autologous HBcAg-expressing LCL of the peripheral blood T cells obtained from the HBVMN patients and HBsAg carriers could be measured. Using autologous HBcAg-expressing LCL as stimulator/target cells for the study of HBcAg-specific cytotoxic T lymphocytes, we found that HBVMN patients had lower cytotoxic activity than did both HBV carriers and HBsAg-/HBsAb+, HBeAg-/HBeAb+ children. From the in vitro cytokine production study of peripheral blood T cells after stimulation with HBcAg, we found that T-helper-cell-1-related IL-2 and IFN-gamma productions were very low in HBVMN patients but T-helper-cell-2-related IL-10 production was higher in HBsAg+/HBeAg+ patients with HBVMN than in those without HBVMN. Based on these findings, we conclude that HBVMN children seem to have an inadequate cellular immune response to HBcAg. PMID- 9200410 TI - Renal angiotensin-converting enzyme localization in diabetic rats and the effect of low protein diet. AB - Recent evidence suggests a role of angiotensin-converting enzyme (ACE) in diabetic nephropathy. The effect of diabetes and low protein diet on renal immunohistochemical ACE localization was studied in streptozotocin-induced DM rats. Immunohistochemical ACE localization was reduced in DM rats, and a low protein diet partially resolved this abnormality while inhibiting the progression of diabetic nephropathy. PMID- 9200412 TI - Selective breeding for high serum IgA levels from noninbred ddY mice: isolation of a strain with an early onset of glomerular IgA deposition. AB - An outbred mouse strain known as ddY has been reported to spontaneously develop, late in life, mesangioproliferative glomerulonephritis with a severe glomerular immunoglobulin A (IgA) deposition that mimics human IgA nephropathy. However, the incidence of the disease in this strain is not very high, probably due to its heterogeneous genetic background. Therefore, we attempted to isolate a strain with a high incidence and an early onset of the disease through selection for high serum IgA from the outbred ddY mice. The selection procedure was successful in increasing the serum IgA level of the selected line and proved effective both in increasing the incidence and in accelerating the onset of the disease. We propose to designate this line of mice 'HIGA', denoting a line with high serum IgA levels. More than half of the mice from the HIGA strain showed a moderate to severe glomerular IgA deposition as early as 25 weeks of age. The severe deposition observed was comparable to that occasionally seen in the original nonselected ddY strain after 40 weeks of age. Thus, we have succeeded in generating a mouse model of IgA nephropathy with a high incidence and an early onset of glomerular IgA deposition. Using light microscopy, progressive and marked mesangial matrix accumulation was shown to develop in HIGA mice. However, they showed only mild proteinuria (100-300 mg/dl) and did not show hematuria. PMID- 9200411 TI - Lysyl oxidase expression and collagen cross-linking during chronic adriamycin nephropathy. AB - Collagen cross-linking induced by lysyl oxidase has been implicated in liver and lung fibrosis. To define the role of this process in kidney fibrosis, we investigated the renal expression of lysyl oxidase and the content in collagen cross-links at various stages of chronic Adriamycin nephropathy in Sprague-Dawley rats. Lysyl oxidase expression was determined by RT-PCR; collagen pyridinium residues, indicating lysyl oxidase induced cross-links, were evaluated by HPLC. These parameters followed a synergic albeit asynchronous outcome: (a) lysyl oxidase mRNA levels in total kidney, glomeruli and medulla from Adriamycin treated rats increased up to 3 times compared to controls between week 8 and 12, then returning within the normal range; (b) the pyridinium residue content did not show any significant difference between Adriamycin-treated and control rats, until diffuse interstitial fibrosis developed (16 weeks), showing at this time a 2- to 3-fold increment. Lysyl oxidase was expressed by several renal cell lines and in tubular-epithelial cells it was up-regulated in vitro by TGF beta-1, a recognized fibrogenetic factor in Adriamycin nephropathy. Our observations demonstrated that an increased expression of lysyl oxidase in the kidney precedes the development of diffuse fibrotic lesions and that, at this stage, collagenic structures contain highly cross-linked components, the final product of lysyl oxidase activity. The evidence of lysyl oxidase up-regulation in tubular epithelial cells by the same factor implicated in Adriamycin toxicity in the kidney suggests a common pathogenetic mechanism. Collagen cross-link formation by lysyl oxidase may be implicated in the pathogenesis of irreversible, fibrotic renal lesions. PMID- 9200414 TI - A young man with acute renal failure and severe loin pain. AB - The first case of exercise-induced acute renal failure (EIARF) is reported measuring the blood flow and arterial resistance in the kidney by pulsed Doppler ultrasound. A 20-year-old Japanese male suffered from severe loin pain and non oliguric acute renal failure after strenuous exercise. Serum myoglobin and creatine phosphokinase were normal and urinary myoglobin was not detectable. The Doppler pattern in several segmental arteries showed a slow end-diastolic velocity (EV) and a high resistance index (RI), indicating increased renal vascular resistance, which suggested severe renal vasoconstriction. Three days later, the EV had apparently increased and the RI normalized in accordance with improvement of renal function. The ultrasound Doppler technique is useful for the detection of a decrease in arterial blood flow on real time and for the diagnosis of EIARF. PMID- 9200413 TI - Extracellular ATP promotes cellular growth of renal inner medullary collecting duct cells mediated via P2u receptors. AB - The present study was undertaken to determine whether extracellular adenosine 5' triphosphate (ATP) promotes cellular proliferation of cultured rat renal inner medullary collecting duct cells. Extracellular ATP increased inositol 1,4,5 triphosphate (IP3) production and cellular free calcium concentration - [Ca2+]i - in a dose-dependent manner. ATP also caused a transient cellular acidification. Extracellular ATP activated mitogen-activated protein (MAP) kinase and [3H]thymidine incorporation in a dose-dependent manner. However, such effects were not obtained with adenosine 5'-diphosphate, adenosine monophosphate, and adenosine. In addition, uridine triphosphate, a P(2u) purinergic agonist, increased IP3 production and activated MAP kinase. 2-Methylthio ATP, a P(2y) purinergic agonist, also increased IP3 production, but did not affect the MAP kinase activity. We also examined the effect of arginine vasopressin on cellular growth. Arginine vasopressin did not alter MAP kinase activity and [3H]thymidine incorporation in cultured rat renal inner medullary collecting duct cells. These results indicate that extracellular ATP activates phospholipase C mediated through P(2u) and P(2y) purinergic receptors and promotes cellular proliferation mediated through P(2u) purinergic receptors in renal inner medullary collecting duct cells. PMID- 9200415 TI - Relapsing membranous nephropathy with a good response to steroid therapy. PMID- 9200417 TI - Urinary excretion of human epidermal growth factor in premature infants requiring assisted ventilation over the first week of life. PMID- 9200418 TI - Therapeutic effect of sarpogrelate, a new 5-hydroxytryptamine receptor 2A antagonist, on diabetic nephropathy and neuropathy. PMID- 9200416 TI - Mesangioproliferative glomerulonephritis in a patient with hepatitis C virus infection and IgA deficiency. PMID- 9200419 TI - Use of parenteral supplements in dialysis patients. PMID- 9200420 TI - High prevalence of hepatitis E virus antibodies in Spanish hemodialysis patients. PMID- 9200421 TI - Do not forget lupus anticoagulant in the era of antiphospholipid antibodies. PMID- 9200422 TI - Neonatal anuria by ACE inhibitors during pregnancy. PMID- 9200423 TI - An unusual case of Wegener's granulomatosis. PMID- 9200426 TI - Measurement of recirculation without peripheral venipuncture. PMID- 9200424 TI - Levels of plasma lipoprotein(a) in continuous ambulatory peritoneal dialysis are not related to peritoneal losses of albumin. PMID- 9200427 TI - A simple assessment of the renal function in geriatric patients. PMID- 9200428 TI - Low prevalence of hepatitis C virus antibodies in a Spanish population with glomerular diseases. PMID- 9200429 TI - Paradoxical and persistent renal impairment in Henoch-Schonlein purpura after high-dose immunoglobulin therapy. PMID- 9200430 TI - Testosterone therapy ameliorates experimental autoimmune encephalomyelitis and induces a T helper 2 bias in the autoantigen-specific T lymphocyte response. AB - Female SJL mice are more susceptible than male mice to experimental autoimmune encephalomyelitis (EAE) induced by myelin basic protein (MBP)-specific T lymphocytes. In the present study, we examined mechanisms involved in this gender related difference in disease susceptibility. MBP-specific T lymphocytes derived from spleens of males during the effector phase of adoptive EAE produced significantly higher levels of IL-10, an anti-inflammatory cytokine in EAE. A protective effect of testosterone was then shown. Females implanted with dihydrotestosterone pellets demonstrated a significantly less severe course of EAE as compared with females implanted with placebo pellets. Finally, MBP specific T lymphocytes derived from dihydrotestosterone-implanted females produced significantly higher levels of IL-10 than those from placebo. Together these data indicate that testosterone exerts a protective effect in EAE that is mediated at least in part by enhanced production of IL-10 by autoantigen-specific T lymphocytes. PMID- 9200431 TI - IL-4 secretion by CD4+ NK1+ T cells induces monocyte chemoattractant protein-1 in early listeriosis. AB - IL-4 is a major promotor of Th2 differentiation and an antagonist of IFN-gamma production. Although experimental listeriosis is characterized by a Th1 response, IL-4-producing cells were detected in spleens of mice promptly after Listeria monocytogenes infection. We identified this early IL-4 as inducer of the chemokine, monocyte chemoattractant protein-1 (MCP-1), which mainly attracts monocytes/macrophages, but not neutrophils. MCP-1-secreting cells were demonstrable in spleens of mice infected with L. monocytogenes, and IL-4 neutralization with anti-IL-4 mAb 11B11 markedly diminished frequencies of MCP-1 producing cells. Cell depletion experiments and studies with gene disruption mutant mice lacking distinct T cell subsets and surface MHC molecules point to CD4+ NK1+ T cells as a cellular source of early IL-4. Since monocyte infiltration to infective foci contributes to early control of listeriosis, our results suggest that IL-4-producing CD4+ NK1+ T cells participate in the innate immune response against L. monocytogenes through MCP-1 induction. PMID- 9200432 TI - The dominant role of bone marrow-derived cells in CTL induction following plasmid DNA immunization at different sites. AB - Although plasmid DNA immunization provides an effective means of inducing CTL responses to an expressed Ag, the mechanism by which CTL precursors are activated remains to be established. Insights could be gained by identifying the cells responsible for Ag presentation when DNA is introduced into different tissue sites. By immunizing parent into F1 bone marrow chimeric mice with an influenza nucleoprotein-expressing plasmid, we have demonstrated that the key cells in this presentation process for both gene gun-mediated epidermal injection and needle intramuscular injection of plasmid DNA are bone marrow derived. Furthermore, as assessed by intramuscular injection, coexpression of nucleoprotein with the costimulatory molecule B7-2, or the cytokines granulocyte-macrophage CSF and IL 12, did not convert nonhemopoietic cells into APCs. Thus, for two distinctly different modes of DNA immunization, in one case with or without coexpressed immunostimulatory factors, the APCs were consistently found to be of hemopoietic origin. PMID- 9200433 TI - Ca2+-dependent, Fas- and perforin-independent apoptotic death of allografted tumor cells by a type of activated macrophage. AB - Both perforin- and Fas ligand (FasL)-deficient CTLs show impaired lytic activity toward most target cells. In this study, we examined whether these molecules could be involved in the cell-to-cell contact-dependent cytotoxicity mediated by macrophages infiltrating into the rejection site of allografted Meth A fibrosarcoma cells. FasL-expressing lymph node cells from MRL-lpr/lpr mice were inactive toward Meth A tumor cells. In C3H/HeJ-gld/gld (abnormal FasL) mice, allograft-induced macrophages (AIM) were cytotoxic against Meth A cells expressing no Fas Ag. Furthermore, allografted Meth A tumor cells were acutely rejected by both C3H/HeJ-gld/gld and control C3H/HeJ mice, indicating that the cytotoxic activity of AIM against Meth A tumor cells was Fas/FasL independent. On the other hand, the cytotoxic activity of AIM against the allografts was dose dependently inhibited by EGTA; and the suppression was restored by the addition of Ca2+, but not Mg2+, implying the involvement of perforin in the cytotoxicity. In the perforin-deficient mice, however, AIM were cytotoxic against Meth A tumor cells; and allografted Meth A tumor cells were acutely rejected by both perforin deficient and control B6 mice, indicating a perforin-independent cytotoxicity. Thus, through a yet unknown mechanism, AIM induced the apoptotic death of allografted Meth A tumor cells. These results indicate that AIM-mediated cytotoxicity against allografted Meth A tumor cells is Ca2+ dependent, and that an attack by AIM results in the apoptotic death of allografted Meth A tumor cells through a third mechanism, one other than the Fas/FasL- and perforin-based pathways. PMID- 9200434 TI - The immunosuppressive metabolite of leflunomide, A77 1726, affects murine T cells through two biochemical mechanisms. AB - The immunosuppressive metabolite of leflunomide, A77 1726, inhibits the enzymatic activity of protein tyrosine kinases and of dihydro-orotic acid dehydrogenase, an enzyme involved in pyrimidine biosynthesis. Here murine CTLL cell lines were studied to determine which of the biochemical targets of A77 1726 was responsible for the observed inhibition of proliferation and cytotoxic activity. At low concentrations of A77 1726, pyrimidine biosynthesis is the target, since inhibition of proliferation correlates with a reduction in pyrimidine NTP levels and is reversed by uridine. At higher concentrations of A77 1726, uridine no longer reverses the inhibition of proliferation even though pyrimidine NTP levels are restored. This second mechanism for inhibiting proliferation is probably inhibition of protein tyrosine kinases, since these higher concentrations of A77 1726 inhibit IL-2-induced tyrosine phosphorylation of Jak1 and Jak3, the protein tyrosine kinases initiating signaling by the IL-2R. Tyrosine phosphorylation of the beta-chain of the IL-2R, which is required for IL-2-driven proliferation, is also inhibited by A77 1726. Cytotoxicity of a CTLL line that overexpresses the Lck protein tyrosine kinase is inhibited by A77 1726; this inhibition is not affected by uridine, but does correlate with inhibition of an Lck in vitro kinase reaction. These studies establish that inhibition of pyrimidine biosynthesis and that of protein tyrosine kinase both contribute to the effects of A77 1726 on CTLL cell lines. PMID- 9200435 TI - IL-12-deficient dendritic cells, generated in the presence of prostaglandin E2, promote type 2 cytokine production in maturing human naive T helper cells. AB - We studied to what extent the presence of an inflammatory mediator PGE2, during the development of dendritic cells (DC) affects their subsequent ability to induce Th1- and Th2-type cytokines in maturing naive Th cells. PGE2 (10(-9)-10( 6) M) did not alter the morphology or the expression of class II MHC and costimulatory molecules on DC obtained from monocytes in the presence of granulocyte-macrophage CSF and IL-4, although at concentrations above 10(-8) M, PGE2 prevented the acquisition of CD1a marker. Both control DC and DC maturing in the presence of PGE2 (PGE2-DC) were potent stimulators of naive Th cells. In contrast to control DC, which produced high amounts of IL-12 and trace amounts of IL-10, PGE2-DC produced no IL-12 and high amounts of IL-10 when stimulated in the absence of PGE2. This distinct cytokine profile of PGE2-DC was stable for at least 48 h of additional culture in the absence of PGE2. Control DC induced the development of Th0-like cells from superantigen-activated naive Th cells, whereas PGE2-DC promoted the development of Th cells that produced high amounts of IL-4 and IL-5. Experiments using IL-12-neutralizing Abs or rIL-12 indicated a crucial role of IL-12 deficiency in the induction of type 2 cytokine profiles. These findings suggest that elevated levels of PGE2 promote type 2 Th responses by stably impairing the ability of maturing DC to produce IL-12. Since type 2 Th responses are protective in several Th1-related autoimmune disorders, PGE2-DC may be considered for use in immunotherapy. PMID- 9200436 TI - Differential expression of transcription directed by a discrete NF-AT binding element from the IL-4 promoter in naive and effector CD4 T cells. AB - Acquiring the ability to selectively produce IFN-gamma or IL-4 is a fundamental property of the immune system and enables T cell subsets (Th1, Th2) to deliver their effector functions. To create an experimental system to examine regulation of critical promoter elements within the IL-4 gene, we have prepared and analyzed transgenic mice expressing the luciferase gene under the control of the distal NF AT binding site from the IL-4 promoter. This site is immediately adjacent to an AP-1 site, and this NF-AT/AP-1 composite site is termed either P1 or PubB. The distal NF-AT site we have used to prepare these reporter transgenic mice lacks the AP-1 binding site but contains the NF-AT binding site (P1(NF-AT)) These transgenic mice were also intercrossed with transgenic mice expressing a single MHC class II-restricted TCR. Expression of transcriptional activity under the control of P1(NF-AT) was observed only in effector T cells, and not naive T cells, after stimulation with Ag or polyclonal stimuli. By contrast, gel mobility shift assays showed that nuclear extracts from both naive and effector T cells contained NF-AT, which could effectively bind to the P1(NF-AT) element. IL-4 stimulated Th2 differentiation did not increase the TCR responsiveness of the P1(NF-AT) element by more than 2-fold but increased production of IL-4 protein by more than 10-fold. These data suggest that factors interacting with the P1(NF-AT) element regulate transcriptional activity in a naive/effector T cell-specific manner but not in a Th1/Th2-specific manner. Th1/Th2-specific regulation of the composite P1 element may result from regulation of transacting factors that bind to the AP-1 portion of this element. PMID- 9200437 TI - Spontaneously colitic C3H/HeJBir mice demonstrate selective antibody reactivity to antigens of the enteric bacterial flora. AB - The idiopathic inflammatory bowel diseases, ulcerative colitis and Crohn's disease, are chronic disorders that appear to arise from an aberrant interaction of environmental, genetic, and immunologic factors. The aim of this study was to examine the immune reactivity of a spontaneously colitic mouse strain, C3H/HeJBir, to epithelial, food, and enteric bacterial Ags. Serum Ab responses of colitic C3H/HeJBir and noncolitic parental C3H/HeJ mice were measured by enhanced chemiluminescence Western blotting. No reactivity to epithelial or food Ags was detected. However, the sera from C3H/HeJBir mice had a reproducible banding pattern on Western blot to bacterial Ags, whereas sera from C3H/HeJ mice did not. Only a small, highly selected number of enteric bacterial Ags were recognized. There were major differences in the degree of recognition of different bacterial strains, marked by remarkably few Abs to Ags of the major anaerobes of the bacterial flora. The serum Abs detected on immunoblot were primarily IgG2a, suggesting a Th1 response. Comparison of sera reactivity to histopathologic severity showed an inverse relationship: one third of young C3H/HeJBir mice during the peak of colitis produced Abs to bacterial Ags, while later in life, when the colitis had resolved, 96% produced Abs. These data are consistent with an abnormal immune reactivity to enteric bacterial flora in C3H/HeJBir mice, a reactivity that is highly selective considering the abundant bacterial Ags present in the colon lumen. We postulate that this reactivity plays a role in the pathogenesis of colitis in these mice. PMID- 9200438 TI - Defective TCR stimulation in anergized type 2 T helper cells correlates with abrogated p56(lck) and ZAP-70 tyrosine kinase activities. AB - Development of IgE-mediated allergic conditions is dependent on the secretion of a Th2 cytokine pattern, including IL-4, IL-5, and IL-13. The induction of anergy would be one mechanism to abrogate cytokine secretion by Th2 cells, which may be pivotal to the allergic response. We demonstrate here that incubation of cloned human CD4+ phospholipase A2 (PLA)-specific Th2 cells with antigenic peptide, in the absence of professional APC, results in a state of nonresponsiveness. The anergic T cells failed to proliferate or secrete IL-4 in response to optimal stimulation with PLA and autologous, professional APC. Secretion of IL-5 and IL 13, however, was only partially inhibited. The anergic state of the Th2 cells was not associated with CD3 or CD28 down-regulation. However, anergy did appear to be closely related to alterations in signaling pathways, mediated through the TCR, of the cells. In contrast to untreated Th2 cells, anergized Th2 cells failed to respond to anti-CD3 mAb with either increased tyrosine kinase activity or increased levels of tyrosine phosphorylation of p56(lck) or ZAP70. A strong and sustained intracellular calcium flux, observed in untreated Th2 cells in response to anti-CD3 mAb, was absent in anergic Th2 cells. Furthermore, the induction of anergy seems to represent an active process, associated with increased levels of basal tyrosine kinase activity, cytokine production, and CD25 up-regulation in anergic Th2 cells. Together, our results indicate that anergy in Th2 cells is associated with defective transmembrane signaling through the TCR. PMID- 9200439 TI - Inactivation of lck and loss of TCR-mediated signaling upon persistent engagement with complexes of peptide:MHC molecules. AB - T cell activation follows recognition of specific peptide:MHC molecule complexes in the context of proper costimulation. The earliest detectable event in T cell activation, within seconds of TCR ligand recognition, is tyrosine phosphorylation of TCR subunits. This causes a cascade of events leading to up-regulation of gene transcription that will drive T cell proliferation and differentiation. Regulation of TCR-mediated signaling upon T cell commitment is unclear. Here, we report that persistent stimulation of T cells, beyond 10 min, correlated with a reversible decrease in tyrosine phosphorylation of T cell lysates that did not affect T cell commitment to proliferation. Loss of Ag-induced tyrosine phosphorylation was not due to lack of Ag presentation, loss of TCR expression, or T cell death, but, rather, it was associated with a lack of TCR subunit tyrosine phosphorylation. We termed this phenomenon TCR desensitization by analogy to the loss of signaling observed in other receptor systems upon persistent engagement with agonist ligands. TCR desensitization correlated with surface reexpression of TCR without concomitant reexpression of coreceptor molecules. Biochemically, TCR desensitization correlated with increased levels of serine-phosphorylated lck, loss of lck kinase activity, and reversible loss of cytosolic lck. Thus, TCR signaling is regulated by desensitization that may be due to serine phosphorylation of lck causing inactivation and loss of this src kinase. This may have important implications by preventing TCR signaling and activation-induced cell death once the T lymphocyte is committed to proliferate. PMID- 9200440 TI - Proto-oncoprotein Vav interacts with c-Cbl in activated thymocytes and peripheral T cells. AB - The molecular adapter c-Cbl is rapidly tyrosine phosphorylated following stimulation through the TCR and associates with Src homology domain-2 (SH2)/SH3 domain-containing adapters such as Grb2, Crk, and Crk-L, which interact with guanine nucleotide exchange factors specific for the Ras family. This suggests that c-Cbl may link TCR activation to molecules that regulate GTP binding proteins. The SH2/SH3-containing protein Vav also contains a guanine nucleotide exchange factor domain, and Vav has a crucial role in thymocyte development and activation of peripheral T cells following stimulation through the TCR. Here we show that Vav and c-Cbl form inducible molecular complexes in TCR-activated murine thymocytes and peripheral T cells as well as pervanadate-treated T cells. Vav/c-Cbl interactions are also detectable in freshly isolated T cells from gene targeted mice that lack the T cell-specific inhibitory receptor CTLA-4, in which c-Cbl is hyperphosphorylated on tyrosine residues. The interaction between Vav and c-Cbl is directly mediated via the SH2 domain of Vav and is dependent on tyrosine phosphorylation of c-Cbl. In addition, we show that the conserved motif Y699 MTP present in c-Cbl is the binding site for the Vav SH2 domain in vitro. These data imply that c-Cbl is a molecular adapter that regulates the function of Vav in thymocytes and peripheral T cells. PMID- 9200441 TI - Immunodominant CD4+ T cell receptor Vbeta repertoires involved in graft-versus host disease responses to minor histocompatibility antigens. AB - In vitro CTL responses to multiple minor histocompatibility Ags (miHA) are governed by immunodominance as demonstrated in the C57BL/6By (B6) anti-BALB.B strain combination. Immunodominance was also demonstrated to be operative in graft-vs-host disease (GVHD) responses against BALB.B-derived miHA following transplantation of B6 T cells into irradiated recipients of both the BALB.B and CXB recombinant inbred strains. The hierarchy of in vivo and in vitro T cell responses to miHA differed. GVHD did not develop in CXBG and CXBK mice, which express immunodominant miHA for CTL generation, whereas disease occurred in the BALB.B, CXBE, CXBI, and CXBJ mouse strains. Previous results demonstrated that B6 CD4+ T cells provide helper function for CD8+ T cells involved in GVHD responses in the BALB.B, CXBE, and CXBI strains. CD4+ T cells alone were mediators of GVHD in all strains except CXBE. This study analyzed the TCR Vbeta repertoires of CD4+ thoracic duct lymphocytes (TDL) collected during the initial stages of GVHD in the B6-->BALB.B and B6-->CXBE strain combinations. Positively selected CD4+ TDL from the B6-->BALB.B (B6(+BALB.B)) combination exhibited marked expansion in the TCR Vbeta6+ and Vbeta8.1/8.2+ families, as well as smaller increases in the Vbeta7 and Vbeta9 families. CD4+ TDL from the B6-->CXBE (B6(+CXBE)) combination displayed expansions in only the Vbeta7+ and Vbeta9+ families. These data suggest that B6 CD4+ T cells can recognize a limited number of immunodominant miHA during GVHD induction and that in both BALB.B and CXBE recipients, the TCR Vbeta repertoires partially overlap. PMID- 9200442 TI - Selective expansion of Vgamma2-Vdelta7 TCR gammadelta cells in C57BL/6 mice is postnatal and extrathymic. AB - Previous studies have shown that TCR-gammadelta cells expressing Vgamma2 region elements are selectively expanded in vivo in C57BL/6 (B6), but not DBA/2, mice. Genetic analysis demonstrated that the expansion of Vgamma2+ was linked to the TCR alphadelta loci, suggesting that a particular Vgamma-Vdelta pair may be necessary for the expansion. In the studies presented here, we find that the expanding TCR gammadelta cells in B6 mice express a Vgamma2+/Vdelta7+ TCR. The Vgamma2-Jgamma and Vdelta7-Ddelta-Jdelta junctional amino acid sequences of these cells display wide variation in length, suggesting that expansion is based on variable region usage and not junctional diversity. The kinetics and dynamics of Vgamma2+/Vdelta7+ T cell expression were studied to determine the biological basis of clonal expansion. Although expression of the Vgamma2+ cells in B6 and DBA/2 neonates was similar, Vgamma2+ cells in the B6 mice expanded fourfold by 4 wk of age, while the expression in DBA/2 mice remained constant. In addition, expansion of the Vgamma2+ cells occurred in athymic nude mice, suggesting that expansion was driven by extrathymic stimuli. Finally, B6 mice housed under germfree conditions expressed expanded levels of Vgamma2+ gammadelta T cells similar to their normally housed counterparts. Thus, expansion and diversification of Vgamma2+/Vdelta7+ T cells are postnatal extrathymic events that do not require microbial antigenic exposure. PMID- 9200443 TI - Regulation of tumor antigen presentation by urocanic acid. AB - Urocanic acid (UCA) accumulates in the epidermis after deamination of histidine. UCA isomerizes from the trans to the cis form upon exposure to environmental UV radiation. Cis-UCA is immunosuppressive in several models. Topically applied cis UCA was reported to enhance the cutaneous tumor yield in chronically UV irradiated mice, suggesting involvement of cis-UCA in photocarcinogenesis. Since Langerhans cells (LC) are capable of presenting tumor-associated Ags (TAA) for primary and secondary tumor-immune responses, we examined the effects of trans- and cis-UCA on LC tumor Ag presentation in a model of immunity to the S1509a spindle cell tumor (H-2a). In this system, induction of immunity requires exposure of LC to granulocyte-macrophage CSF. Naive CAF1 (H-2(a/d)) mice were immunized against S1509a by injection with granulocyte-macrophage CSF-exposed and TAA-pulsed epidermal cells (EC), as assessed by growth inhibition of inoculated tumor cells. Incubation of EC in cis-, but not trans-UCA completely inhibited Ag presentation in this system. Neither histamine antagonists nor indomethacin reversed these effects of cis-UCA. The ability of trans- and cis-UCA to modulate EC presentation of TAA for secondary immune responses was also examined. EC were pulsed with TAA in vitro and then injected into hind footpads of tumor-immune mice. After 24 h, footpad swelling was assessed as a measure of delayed-type hypersensitivity. Incubation with cis-, but again not trans-UCA before TAA exposure significantly inhibited elicitation of delayed-type hypersensitivity. These data indicate that cis-UCA may be an important regulator of LC Ag presenting function in tumor-immune responses, and thus may play a role in photocarcinogenesis. PMID- 9200444 TI - Propionibacterium acnes treatment diminishes CD4+ NK1.1+ T cells but induces type I T cells in the liver by induction of IL-12 and IL-18 production from Kupffer cells. AB - LPS injection into normal mice does not induce liver injury, while the same treatment of Propionibacterium acnes-primed mice induces severe liver injury, indicating that P. acnes treatment renders the mice susceptible to LPS. Since IFN gamma sensitizes macrophages to LPS, we investigated the mechanism of induction and activation of IFN-gamma-producing (type 1) T cells by P. acnes. Twenty percent of liver lymphocytes of C57BL/6 mice are CD4+ NK1.1+ T cells that promptly produce IL-4 in response to anti-CD3 in vitro. However, P. acnes treatment diminished these lymphocytes. Therefore, liver lymphocytes from P. acnes-primed mice showed reduced IL-4 production. Furthermore, P. acnes treatment induced CD4- type 1 T cells in the liver. Isolated P. acnes-elicited Kupffer cells produced IL-12 and to a lesser degree IL-18 in vitro. Injection of anti-IL 12 Ab totally abrogated these actions of P. acnes, while injection of anti-IL-18 Ab caused only partial abrogation. Thus, administration of P. acnes diminished CD4+ NK1.1+ T cells, but induced type 1 T cells in the liver by induction of IL 12 and IL-18 production. Injection of IL-12 (approximately 1,000 ng) dose dependently diminished CD4+ NK1.1+ T cells, but induced type 1 T cells. In contrast, injection of IL-18 (approximately 1,000 ng) failed, although injection of a much larger dose of IL-18 (10,000 ng) or IL-18 (approximately 1,000 ng) with suboptimal doses of IL-12 (1-100 ng) diminished CD4+ NK1.1+ T cells in a dose dependent manner. Thus, P. acnes treatment renders the mice highly susceptible to LPS by induction and activation of type 1 T cells. PMID- 9200445 TI - Inhibition of primary and recall allergen-specific T helper cell type 2-mediated responses by a T helper cell type 1 stimulus. AB - Allergic responses are characterized by the production of Ag-specific IgE Abs that are dependent upon Th2-mediated T cell help. We determined whether heat killed Brucella abortus (BA), an inducer of Th1 responses, could influence the allergic Th2-mediated IgE response to OVA adsorbed to alum (O/A). BA plus O/A, but not O/A alone, induced high levels of mRNA for IFN-gamma and IL-12 promptly after injection. Furthermore, initial treatment with BA plus O/A rendered both BALB/c and C57Bl/6 mice incapable of mounting high IgE responses even after repeated challenges with allergen alone. Long term abrogation of anti-OVA IgE correlated with an increased frequency of IFN-gamma-secreting OVA-specific cells and a decreased frequency of IL-4-secreting OVA-specific cells. Initial treatment with anti-IL-12 prevented BA-induced early IFN-gamma production and secondary IgG2a responses, but did not abrogate IgE suppression. Additionally, secondary OVA-specific IgE responses were down-regulated by BA conjugated to OVA or by BA given with O/A. BA-induced down-regulation of secondary IgE responses was associated with increased frequency of Ag-specific IFN-gamma-secreting cells. These results suggest the possibility that even recall Th2-mediated immune responses can be attenuated if Ag is given with a carrier or adjuvant that induces potent Th1-promoting cytokines. PMID- 9200446 TI - TGF-beta down-regulates stromal IL-7 secretion and inhibits proliferation of human B cell precursors. AB - Development of lymphoid progenitors in vivo requires interaction with a bone marrow stromal microenvironment containing multiple cytokines involved in the development of nonlymphoid hemopoietic lineages. We tested the effect of one such cytokine, TGF-beta, on the proliferation of early human clonogenic lymphoid progenitors using a stroma-dependent in vitro culture system. TGF-beta caused a dose-dependent inhibition of lymphoid progenitor colonies that was reversible at low TGF-beta doses by addition of exogenous IL-7 to the cultures. IL-7 was unable to reverse the inhibitory effect of higher TGF-beta concentrations or inhibition caused by IL-1alpha, IL-4, or TNF-alpha. Stromal IL-7 mRNA expression and protein secretion were markedly down-regulated by TGF-beta, suggesting that inhibition of stromal IL-7 secretion partially accounts for the inhibitory effect of TGF-beta on lymphopoiesis in this culture system. It is likely that higher TGF-beta concentrations do not inhibit lymphopoiesis by down-regulating IL-7 receptor expression, since this cytokine did not reduce IL-7R alpha or gamma c mRNA levels in normal B cell precursors. Since direct stromal contact is required for in vitro lymphopoiesis, the potential regulation of the IL-7 pathway by cell adhesion was examined. Adhesion of human B cell precursors to stroma did not alter stromal IL-7 expression or expression of IL-7R alpha or gamma c-chains by B cell precursors. These results indicate that TGF-beta is a significant negative regulator of stroma-dependent proliferation of early human lymphoid progenitors and acts in part by down-regulating stromal IL-7 secretion. PMID- 9200447 TI - Bcl-2 expression in target cells leads to functional inhibition of caspase-3 protease family in human NK and lymphokine-activated killer cell granule-mediated apoptosis. AB - In the granule exocytosis pathway of cell-mediated cytotoxicity, rapid apoptotic nuclear damage in target cells has been unequivocally linked to granzyme B activity. Direct cleavage and activation of caspase-3 and related proteases by granzyme B have been identified as a central event in apoptosis induction by cytotoxic granules. The Bcl-2 oncoprotein has been recently shown to act at the level or upstream of caspase-3 family activation to inhibit apoptosis induced by various stimuli including Fas ligation, an alternative cell-mediated lytic pathway. In this study, we have investigated whether activation of this caspase family by granzyme B, during human NK and lymphokine-activated killer cell granule-mediated apoptosis, could be influenced by Bcl-2 expression. Bcl-2 overexpressing clones were generated from parental K562 and U937 cell lines (K6 and U4 clones, respectively). Bcl-2 expression abrogated early 125I-DNA release and DNA fragmentation, these defects being compensated for by extended incubation times. Cleavage of poly(ADP-ribose) polymerase, a specific caspase-3 family substrate, was detected in parental K562 cells exposed to lymphokine-activated killer effectors but not in K6 targets, indicating that caspase-3 and related proteases function was inhibited by Bcl-2. Functional inhibition of caspase-3 family with benzyloxycarbonyl-Asp-Glu-Val-Asp(OMe) fluoromethylketone led to similar consequences on apoptotic nuclear events as for Bcl-2 expression. Thus, Bcl-2 antagonizes granzyme B-mediated apoptosis by a mechanism that interferes with caspase-3 activity. Finally, Bcl-2 expression or the Asp-Glu-Val-Asp peptide was much less efficient in preventing phosphatidylserine externalization, suggesting that despite impaired nuclear apoptosis, immediate recognition and elimination of Bcl-2-expressing cells by tissue phagocytes should remain partly unaffected. PMID- 9200448 TI - xid affects events leading to B cell cycle entry. AB - X-linked agammaglobulinemia patients and X-linked immunodeficient (xid) mice possess mutations in the Bruton's tyrosine kinase (Btk kinase) gene and display defects in B cell development and activation by sIg cross-linking. Btk is an early activation kinase in sIg-cross-linked B cells. xid does not ablate Btk protein kinase activity, and immediate signal transduction events, such as tyrosine phosphorylation, occur in sIg-activated xid B cells. These cells do not subsequently progress into cell division and have a high rate of apoptosis, which has been shown to correlate with an absence of sIg-mediated induction of the bcl xL protein. To establish the point where Btk activity is critical for progression beyond immediate signaling, we examined early and late events in sIg-cross-linked xid B cells. Induction of proto-oncogenes and nuclear factors occurred normally in xid cells. However, induction of cyclins and increased GAPDH mRNA was not observed in xid cells. Degradation of the cyclin inhibitor p27Kip1 occurred normally in xid cells. After 24 h of culture with anti-mu, the remaining live, nonapoptotic xid cells were enlarged, viable, and primed for subsequent stimulation by LPS. Our data suggest that the Btk kinase is not essential for several G1 events and that the failure of sIg-activated xid B cells to enter cell cycle correlates with a defect of cyclin induction. Moreover, these data suggest that Btk is important not only for immediate events following B cell activation and control of apoptosis but also for subsequent events leading to cyclin activation. PMID- 9200449 TI - Interaction of CTLA-4 with the clathrin-associated protein AP50 results in ligand independent endocytosis that limits cell surface expression. AB - CTLA-4 is a lymphocyte cell surface receptor expressed by activated T cells that functions to down-regulate T cell responses induced by TCR and CD28 stimulation. Since CTLA-4 competes with CD28 for binding to the common ligands B7-1 and B7-2, the level of CTLA-4 surface expression is likely to play an important role in its ability to inhibit CD28-dependent T cell activation. The factors that regulate these levels are poorly understood. Recent studies have revealed that following T cell activation, the majority of CTLA-4 is localized intracellularly rather than on the cell surface, and surface CTLA-4 is rapidly reinternalized. In this study, we investigate the molecular mechanism underlying the rapid clearance of CTLA-4 from the cell surface. The data demonstrate that cell surface CTLA-4 is endocytosed into clathrin-coated vesicles even in the absence of ligand. The targeting of CTLA-4 to clathrin-coated vesicles is mediated by the clathrin associated adaptor complex AP-2. The cytoplasmic domain of CTLA-4 was found to specifically bind to AP50, the medium chain subunit of AP-2 in both yeast two hybrid and coimmunoprecipitation assays. The interaction requires the peptide sequence 199-GVYVKM-204 in the cytoplasmic tail of CTLA-4. Mutation of the CTLA-4 amino acid residue Y201 abrogates the interaction with AP50, resulting in the accumulation of CTLA-4 at the cell surface. Together these data suggest that the interaction of CTLA-4 with AP50 plays an important role in regulating the cell surface expression of CTLA-4. PMID- 9200450 TI - Direct MHC class I complementary DNA transfer to thymus induces donor-specific unresponsiveness, which involves multiple immunologic mechanisms. AB - Our purposes were 1) to determine whether direct transfer of cDNA encoding allogeneic MHC class I Ag to the rat thymus would be capable of inducing donor specific unresponsiveness and 2) to study the immunologic mechanism of this effect. Plasmid DNA encoding donor strain (ACI-RT1.Aa) MHC class I Ag was directly injected into Lewis (RT1(l)) rat recipient thymus 7 to 10 days before ACI liver transplantation. A single dose of anti-lymphocyte serum was given i.p. on the day of thymic injection. Rats injected intrathymically with plasmid DNA and treated with anti-lymphocyte serum demonstrated prolonged survival in 9 of 13 rats (>100 days). PCR was used to demonstrate that RT1.Aa cDNA was expressed in thymus transiently and later appeared in spleen. CTL limiting dilution assays showed that CTL precursor frequency was decreased in tolerant liver recipients. To test the hypothesis of clonal deletion vs anergy, CTL limiting dilution assays cultures were restimulated with donor cells and IL-2 to reverse anergy. Restimulation caused CTL precursor frequency to return to near normal in only one of five tolerant rats, suggesting clonal deletion or a dense anergic state. Passive transfer of splenocytes from tolerant rats to naive recipients prolonged cardiac allograft survival, suggesting that suppressor-type cells may also contribute to thymic tolerance in our model. In summary, our data suggest that donor MHC class I Ag expressed in thymus by direct DNA injection, followed by liver allografting, results in donor-specific unresponsiveness. The mechanism of this effect is complex, involving multiple immunologic mechanisms. PMID- 9200451 TI - Requirement of CD4 T cells for skin graft rejection against thymus leukemia (TL) antigen and multiple epitopes on the TL molecule recognized by CD4 T cells. AB - By selective depletion of CD4 and CD8 T cells in vivo using the respective mAbs, we demonstrate that CD4 T cells are necessary for skin graft rejection against thymus leukemia (TL) Ag. The skin expressing T3b-TL Ag from transgenic C3H Tg.Con.3-1 mice given chimeric H-2Kb/T3b-TL gene was rejected when grafted onto C3H/He recipient mice. Depletion of CD4, but not of CD8, T cells blocked rejection. CD8 CTL were generated in MEM (control)-treated C3H/He recipient mice, while Thy-1+ CD4- CD8- CTL were generated in CD8-depleted recipient mice after rejection. However, no CTL were generated in CD4-depleted or both CD4- and CD8 depleted recipient mice. Thus, the generation of both CD8 and Thy-1+ CD4- CD8- CTL was dependent on CD4 T cells. Ab blocking indicated that both CD8 and Thy-1+ CD4- CD8- CTL were TCR alphabeta and recognized TL Ag. We furthermore demonstrated that CD4 T cells in spleen cells from C3H/He mice that had rejected C3H Tg.Con.3-1 skin showed a weak, but significant, proliferative response to in vitro stimulation with mitomycin C-treated C3H Tg.Con.3-1 spleen cells. Analysis of the reactivity of bulk CD4 T cell lines to 73 synthetic overlapping peptides encompassing the entire T3b-TL molecule showed that CD4 T cells recognized multiple epitopes on the T3b-TL molecule in an APC-dependent manner. PMID- 9200452 TI - In vivo mechanism by which leflunomide controls lymphoproliferative and autoimmune disease in MRL/MpJ-lpr/lpr mice. AB - Two activities have been identified for the immunosuppressive metabolite of leflunomide, A77 1726: inhibition of dihydroorotate dehydrogenase (DHO-DHase), an enzyme involved in the biosynthesis of pyrimidine nucleotides (PyN); and inhibition of protein tyrosine kinases. The in vitro potency of A77 1726 as a DHO DHase inhibitor is reported to be 10- to 500-fold greater than as a tyrosine kinase inhibitor. These observations suggested that the immunosuppressive efficacy of leflunomide in vivo is related to inhibition of DHO-DHase. However, observations that patients with disorders in the PyN synthetic pathway are not overtly immunodeficient militate against this hypothesis. We investigated the effects of leflunomide in vivo and report that amelioration of lymphoproliferative and autoimmune diseases in MRL/MpJ-lpr/lpr (lpr/lpr) mice by leflunomide is not accompanied by reduced PyN concentrations in lymph node cells. Our hypothesis that lymphocytes could salvage serum uridine to counter the effects of reduced PyN synthesis in vivo was supported by in vitro studies. Finally, we observed that amelioration of disease correlated with a reduction of tyrosine phosphorylated proteins in lymph node cells of lpr/lpr mice. These observations suggest that the primary mechanism by which leflunomide prevents autoimmune and lymphoproliferative diseases in lpr/lpr mice is not depletion of PyN, but correlates with reduced tyrosine phosphorylation concentrations in lymph node cells. PMID- 9200453 TI - TGF-beta inhibits IL-2-induced tyrosine phosphorylation and activation of Jak-1 and Stat 5 in T lymphocytes. AB - Signaling through IL-2R, IL-2 induces tyrosine phosphorylation and activation of Jak-1 and Jak-3 kinases and Stat 3 and Stat 5 transcription factors leading to cell cycle progression of activated T cells from G1 to S phase. TGF-beta is an immunosuppressive cytokine, which inhibits T cell proliferation at G1 to S phase transition. We examined the effect of TGF-beta on IL-2R signal transduction pathway in activated T cells. We show here that treatment of activated T cells with TGF-beta inhibited IL-2-induced tyrosine phosphorylation and activation of Jak-1 and Stat 5 but not Jak-3 and Stat 3. TGF-beta also inhibited IL-2-induced expression of alpha- and beta-chains of IL-2R and induced apoptotic cell death in T cells. These results suggest that TGF-beta-induced growth arrest and apoptosis are associated with the modulation of IL-2-induced activation of Jak-Stat pathway in T cells. PMID- 9200454 TI - CD38 ligation in human B cell progenitors triggers tyrosine phosphorylation of CD19 and association of CD19 with lyn and phosphatidylinositol 3-kinase. AB - CD38 is a 45-kDa transmembrane glycoprotein highly expressed in lymphoid progenitors. Ligation of CD38 with specific Abs inhibits growth and induces apoptosis in human immature B cells. CD38 ligation also triggers tyrosine phosphorylation of syk, c-cbl, and phospholipase C-gamma and activates phosphatidylinositol 3-kinase (PI3-K). In the present study, we investigated whether the cell surface membrane molecules used in B cell receptor-mediated signaling, such as Ig alpha, Ig beta, and CD19, could be involved in the CD38 mediated signaling cascade. In the B cell receptor-negative immature B cell lines RS4;11, 380, and REH, Ig alpha and Ig beta were expressed exclusively in the cytoplasm and were not tyrosine phosphorylated after CD38 ligation. By contrast, CD19 was markedly tyrosine phosphorylated and was associated with lyn and PI3-K. PI3-K activation appears to be directly linked to the growth-arresting effects of CD38 ligation, which are reduced by PI3-K inhibitors. Ligation of either CD38 or CD19 resulted in a similar pattern of protein tyrosine phosphorylation; both signaling pathways caused tyrosine phosphorylation of c-cbl. Levels of CD38 surface expression were not affected by prolonged incubation with anti-CD19 Ab, while CD19 expression markedly decreased. These results indicate that CD19 is a major component of the CD38 signaling cascade in B cell precursors, serving as a cell surface membrane docking site for cytoplasmic kinases. CD38 and CD19 are not physically linked, but activate an overlapping set of kinases in human immature B cells. PMID- 9200456 TI - Expression and role in apoptosis of the alpha- and beta-chains of the IFN-gamma receptor on human Th1 and Th2 clones. AB - The mRNA and protein expression of the alpha- and beta-chains of IFN-gammaR were evaluated on a panel of human Th1 and Th2 clones. When cultured in IL-2 conditioned medium, both types of clones expressed mRNA for the alpha- and beta chains, and both chains were present in the cytoplasm. Membrane expression of the alpha-chain was higher on Th2 than on Th1, whereas the beta-chain was poorly expressed on both types but increased following IL-2 withdrawal or PHA stimulation. In addition, both types of clones overexpressed MHC class I glycoproteins following IFN-gammaR triggering by exogenous IFN-gamma, although the kinetics was slower in Th1, and this exposure induced mRNA for IRF-1. When their TCR was triggered in the absence of APC, Th1 only underwent apoptosis. This activation-induced apoptosis was prevented by blocking of the alpha-chain or by IFN-gamma neutralization. Addition of IFN-gamma triggered the apoptosis of Th2 clones. Apoptosis of both types of clones was mediated by autocrine or exogenous IFN-gamma through the up-regulation of Fas-L expression, since anti-IFN-gammaR alpha mAb inhibited its expression on Th1 and exogenous IFN-gamma increased its expression on Th2. These results indicate that activated human Th1 and Th2 lymphocytes express IFN-gammaR alpha- and beta-chains and are both sensitive to signals provided by IFN-gamma. Data also suggest that IFN-gamma is critical for switching off their responses. PMID- 9200455 TI - CD38 ligation results in activation of the Raf-1/mitogen-activated protein kinase and the CD3-zeta/zeta-associated protein-70 signaling pathways in Jurkat T lymphocytes. AB - CD38 ligation with the specific mAb IB4 induced early and late signaling events in Jurkat T cells, as judged by the transient induction of tyrosine phosphorylation of phospholipase C-gamma1, c-Cbl, zeta-associated protein (ZAP) 70, Shc, extracellular signal-regulated protein kinase-2 (Erk-2) as mitogen activated protein (MAP) kinase, and increased expression of the activation Ag CD69. In addition, CD38 ligation induced Ras-dependent events such as Erk-2 mobility shift and increased Erk-2 kinase activity. Further evidence that Erk-2 activation is regulated by CD38 ligation was obtained indirectly with the observed induction of Raf-1, Lck, and Sos-1 mobility shifts, processes that are believed to be dependent, at least in part, on MAP kinase activation. Using a protein tyrosine kinase inhibitor, herbimycin A, or a protein kinase C inhibitor, Ro-31-8220, we found that the anti-CD38-induced Erk-2 activation is both protein tyrosine kinase and protein kinase C dependent. CD38 ligation also resulted in increased CD3-zeta tyrosine phosphorylation and its association with ZAP-70. CD38 ligation in a Jurkat Lck-deficient mutant, JCam1, failed to induce substrate tyrosine phosphorylation and activation of Erk-2. These data indicated that in Jurkat T cells, CD38 receptor triggering results in Lck-regulated activation of both Raf-1/MAP kinase and CD3-zeta/ZAP-70/phospholipase C-gamma1 signaling pathways. PMID- 9200457 TI - Antigen specificity of dual reactive T hybridomas determines the requirement for CD40 ligand-CD40 interactions. AB - CD40 ligand (CD40L) expression on T cells is known to play a crucial role in B cell responses. Some evidence also supports a role for CD40L-CD40 interactions in T cell responses, at least in vivo. Whether the T cell requirement for these interactions is an invariable finding, however, is less clear. Here, we provide evidence that the Ag specificity of T cells influences the requirement for CD40L. T cell hybridomas with dual reactivity for two different Ags, allo-H2-Ap and Mls(a) superantigens, display a differential requirement for CD40L expression. Whereas the response to splenic APC expressing Mls(a) Ags requires CD40L expression, the response to alloantigen-bearing APC does not. The requirement for CD40L expression for the Mls(a) response appears to reflect a strong dependence of this response on ICAM-1 (intercellular adhesion molecule-1) and the ability of CD40-mediated signals to regulate ICAM-1 expression. These findings demonstrate that CD40L-CD40-mediated cross-talk is important for some but not all T cell responses and is influenced by both the type of Ag recognized and the type of APC. PMID- 9200458 TI - Fc epsilonRI gamma can support T cell development and function in mice lacking endogenous TCR zeta-chain. AB - Fc epsilonRI gamma (Fc gamma) is a member of the zeta family of signal transducing molecules that function as components of both the TCR and Fc receptors (FcR). While the majority of thymocytes and T cells express TCRs containing zeta-chain homodimers, certain unique populations of T cells express TCRs that contain both zeta and Fc gamma. To examine the ability of Fc gamma to substitute for zeta-chain in T cell development and function, we introduced a transgene encoding Fc gamma into mice made genetically deficient for zeta-chain (zeta(e)-/-). Analysis of thymocyte development in zeta(e)-/-;Fc gamma Tg mice demonstrated that Fc gamma was able to support the maturation of both gammadelta TCR+ and alphabeta TCR+ T cells. However, positive selection of alphabeta TCR+ thymocytes was less efficient in zeta(e)-/-;Fc gamma Tg mice than in zeta(e)-/- mice reconstituted with zeta-chain. This difference may be due to the fact that Fc gamma contains a single immunoreceptor tyrosine-based activation motif (ITAM) whereas zeta-chain contains three ITAMs. Interestingly, the peripheral T cells that develop in zeta(e)-/- mice reconstituted with Fc gamma are functional and respond to TCR-specific stimuli. These data suggest that Fc gamma and zeta are interchangeable in their ability to mediate T cell development and function, however zeta-chain is more efficient at promoting positive selection and T cell maturation. The difference in efficiency between zeta and Fc gamma may be responsible in part for the unusual developmental and functional properties of T cells that constitutively express Fc gamma as a signaling component of their TCRs. PMID- 9200460 TI - Structural requirements for assembly of dimeric IgA probed by site-directed mutagenesis of J chain and a cysteine residue of the alpha-chain CH2 domain. AB - The structural features of J chain required for interaction with IgA in IgA dimer assembly were investigated by coexpression of wild-type and mutant forms of J chain with IgA1 in CHO cells. With wild-type J chain, a mixture of J chain containing dimers and monomers was secreted. Substitution of Cys14 of J chain with Ser resulted in expression of only monomer IgA covalently associated with J chain. Similarly, mutation of Cys68 to Ser also resulted in expression predominantly of a monomer IgA-J chain species. These results suggest that Cys14 and Cys68 play critical roles in formation of J chain-containing IgA dimers, with each forming a disulfide bridge to an IgA monomer. Substitution of Asn48 with Ala, to prevent attachment of N-linked carbohydrate to J chain, also resulted in markedly reduced dimer assembly, suggesting a requirement for the sugar moiety in J chain function. We also mutated Cys311 on the C alpha2 domain of the IgA heavy chain to Ser. When coexpressed with wild-type J chain, this mutant was still capable of forming dimers, indicating that this residue was not involved in dimerization. Taken together, our results are consistent with an arrangement in which IgA monomers are linked end-to-end with J chain interposed. PMID- 9200459 TI - B cell antigen receptor desensitization: disruption of receptor coupling to tyrosine kinase activation. AB - Antigen binding to the B cell receptor (BCR) induces receptor desensitization, a condition characterized by cellular unresponsiveness to subsequent Ag stimulation despite the continued ability to bind Ag. To better understand the molecular mechanism of this unresponsiveness, we have used complementary lymphoma (K46 mu) and Ig transgenic (3-83 mu delta) mouse models to study regulation of BCR signaling. Our findings in the lymphoma model show that an initial Ag encounter renders receptors unresponsive to subsequent Ag challenge, as measured by their inability to mobilize Ca2+ and to mediate phosphorylation of receptor-proximal kinases, including Lyn, Blk, and Syk. Most importantly, the Ig alpha and Ig beta components of desensitized receptors are not phosphorylated, and receptor associated kinases are not activated upon Ag challenge. The molecular defect does not appear to result from Lyn inactivation, sequestration, or repression, since Lyn from desensitized cell lysates is activated in vitro by synthetic doubly phosphorylated immunoreceptor tyrosine-based activation motif peptides. A similar deficit in Ag-induced receptor phosphorylation was observed in desensitized B cells from 3-83 mu delta transgenic mice. These studies indicate that Ag receptor desensitization reflects an inability to initiate activation of receptor associated kinases that normally phosphorylate receptor Ig alphabeta subunits, leading to signal propagation. PMID- 9200461 TI - Structure and genomic organization of a second class of immunoglobulin light chain genes in the channel catfish. AB - Earlier studies distinguished two classes of catfish light (L) chain (designated F and G). The cDNA structure and genomic organization of G L chain gene clusters has also been characterized previously. In this study, full length cDNA encoding F L chain was derived using PCR strategies based on the determined amino-terminal protein sequence. The encoded V region is readily delineated into framework regions (FR) and complementarity-determining regions (CDR). Multiple sequence alignments indicate that the F V(L) is closely related to kappa gene families. The F C(L) cannot be generally classified but it is structurally distinct from the C(L) regions of G: the amino acid sequence similarity is <35%. cDNA sequences representing processed sterile F transcripts of different loci were identified. Each sequence begins within the J(L) recombination signal sequence and extends downstream through the I(L)-C(L) segments. Genomic blots hybridized with C(L) probes indicate that there are at least 50 different C(L) segments. Based upon V(L) hybridization studies, different families of V(L) segments appear to be associated with closely related F C(L) segments. In characterized genomic clones, F gene segments are arranged in closely linked clusters with single copies of V(L), J(L), and C(L) segments within each cluster. The V(L) segments are located in opposite transcriptional polarity relative to the J(L) and C(L) segments, which indicates that V(L) segments rearrange by inversion. These combined studies establish that two structurally distinct classes of L chains are present in teleost fish and that both of the L chain classes evolved within a common organizational pattern of clustered segmental genes. PMID- 9200462 TI - Isolation and characterization of 2'-fluoro-, 2'-amino-, and 2'-fluoro-/amino modified RNA ligands to human IFN-gamma that inhibit receptor binding. AB - CD4+ Th cells produce cytokines that play a pivotal role in the induction and regulation of cell-mediated and humoral immunity. Th1 cells, characterized by their secretion of IFN-gamma, induce macrophage cytotoxicity, delayed hypersensitivity, and enhanced cellular immunity. Secretion of IFN-gamma may even suppress Th2-enhanced humoral immunity. A counterproductive Th1 response and concomitant secretion of IFN-gamma may result in inflammatory and autoimmune diseases. IFN-gamma regulation of T cell function has potential for therapeutic intervention. To isolate high affinity oligonucleotide inhibitors of IFN-gamma activity, combinatorial libraries of RNA molecules modified at the 2' position of pyrimidine nucleotides with fluoro (F), amino (NH2), or a mixture of F and NH2 (2'-F/NH2) were screened using the SELEX (systematic evolution of ligands by exponential enrichment) combinatorial chemistry process. Each modified library of RNA molecules provides an expanded repertoire of molecules with increased structural diversity and unique binding properties. This added diversity increases the possibility of isolating molecules with the desired functional properties. These RNAs modified at the 2' position have also been shown to be nuclease resistant. High affinity ligands to human IFN-gamma from each modified library were isolated and characterized. The K(d)s of these ligands were determined and their secondary structures were predicted. The specificity of these ligands for IFN-gamma binding was confirmed, and their ability to inhibit binding of IFN-gamma to its receptor on A549 human lung carcinoma cells was determined. A 2'-NH2-modified ligand (2'-NH2-30) is described that binds IFN gamma with high affinity and inhibits IFN-gamma-induced expression of MHC class I and ICAM-1 by human myeloid leukemia cells. PMID- 9200463 TI - The alpha subunit cytoplasmic domain regulates the assembly and adhesiveness of integrin lymphocyte function-associated antigen-1. AB - The integrin LFA-1 mediates activation-dependent leukocyte adhesion. The beta subunit cytoplasmic domain has been demonstrated previously to modulate the adhesiveness of LFA-1. To investigate whether the alpha subunit cytoplasmic domain is also involved in the regulation of LFA-1-adhesive function, we stably expressed cytoplasmic domain truncated forms of the alpha subunit in a Jurkat mutant (Jurkat-beta2.7) deficient in the endogenous LFA-1 alpha subunit and in K562 cells. Clones expressing similar levels of cell surface LFA-1 were tested for their ability to bind to immobilized ICAM-1. Truncation of the alpha subunit cytoplasmic domain before, but not after, the conserved GFFKR sequence motif resulted in constitutive ICAM-1 binding of both Jurkat-beta2.7 and K562 transfectants. However, truncation after the GFFKR motif reduced sensitivity to stimulation by PMA or stimulatory Abs. Internal deletion of the GFFKR motif, or point mutations of the Gly (G), the two Phe (F), or the Arg (R) in the GFFKR motif to Ala (A) rendered LFA-1 constitutively active. Mutation of the Lys (K) did not affect LFA-1 adhesion to ICAM-1. These findings indicate that the GFFKR motif maintains the low adhesive state of LFA-1, possibly by restraining the receptor conformation. We further demonstrate that the alpha subunit cytoplasmic domain and the conserved GFFKR motif are also required for efficient formation of LFA-1 alphabeta heterodimers. PMID- 9200464 TI - Refocusing neutralizing antibody response by targeted dampening of an immunodominant epitope. AB - Immunodominant epitopes are known to suppress a primary immune response to other antigenic determinants by a number of mechanisms. Many pathogens have used this strategy to subvert the immune response and may be a mechanism responsible for limited vaccine efficacies. HIV-1 vaccine efficacy appears to be complicated similarly by a limited, immunodominant, isolate-restricted immune response generally directed toward determinants in the third variable domain (V3) of the major envelope glycoprotein, gp120. To overcome this problem, we have investigated an approach based on masking the V3 domain through addition of N linked carbohydrate and reduction in net positive charge. N-linked modified gp120s were expressed by recombinant vaccinia virus and used to immunize guinea pigs by infection and protein boosting. This modification resulted in variable site-specific glycosylation and antigenic dampening, without loss of gp120/CD4 binding or virus neutralization. Most importantly, V3 epitope dampening shifted the dominant type-specific neutralizing Ab response away from V3 to an epitope in the first variable domain (V1) of gp120. Interestingly, in the presence of V3 dampening V1 changes from an immunodominant non-neutralizing epitope to a primary neutralizing epitope with broader neutralizing properties. In addition, Ab responses were also observed to conserved domains in C1 and C5. These results suggest that selective epitope dampening can lead to qualitative shifts in the immune response resulting in second order neutralizing responses that may prove useful in the fine manipulation of the immune response and in the development of more broadly protective vaccines and therapeutic strategies. PMID- 9200465 TI - Human alveolar T lymphocyte responses to Mycobacterium tuberculosis antigens: role for CD4+ and CD8+ cytotoxic T cells and relative resistance of alveolar macrophages to lysis. AB - T cell-mediated cytotoxicity against Mycobacterium tuberculosis (MTB)-infected macrophages may be a major mechanism of specific host defense, but little is known about such activities in the lung. Thus, the capacity of alveolar lymphocyte MTB-specific cell lines (AL) and alveolar macrophages (AM) from tuberculin skin test-positive healthy subjects to serve as CTL and target cells, respectively, in response to MTB (H37Ra) or purified protein derivative (PPD) was investigated. Mycobacterial Ag-pulsed AM were targets of blood CTL activity at E:T ratios of > or = 30:1 (51Cr release assay), but were significantly more resistant to cytotoxicity than autologous blood monocytes. PPD- plus IL-2 expanded AL and blood lymphocytes were cytotoxic for autologous mycobacterium stimulated monocytes at E:T ratios of > or = 10:1. The CTL activity of lymphocytes expanded with PPD was predominantly class II MHC restricted, whereas the CTL activity of lymphocytes expanded with PPD plus IL-2 was both class I and class II MHC restricted. Both CD4+ and CD8+ T cells were enriched in BL and AL expanded with PPD and IL-2, and both subsets had mycobacterium-specific CTL activity. Such novel cytotoxic responses by CD4+ and CD8+ T cells may be a major mechanism of defense against MTB at the site of disease activity. PMID- 9200466 TI - Production of IL-18 (IFN-gamma-inducing factor) messenger RNA and functional protein by murine keratinocytes. AB - Recently, the novel cytokine IL-18 (IFN-gamma-inducing factor) has been described as a growth and differentiation factor for Th1 cells. Epidermal keratinocytes (KC) are known to direct T cell education by production of cytokines. Therefore, expression of IL-18 was sought in KC. Epidermal RNA was analyzed following stimulation with contact sensitizers or controls for IL-18 mRNA expression by semiquantitative reverse transcription-PCR. Constitutive expression of IL-18 mRNA was low in untreated epidermal cells (EC), but early up-regulation of IL-18 mRNA signals was detected following application of a contact allergen in vivo. The peak strength of IL-18 signals was observed within 4 to 6 h following stimulation with an allergen. Application of an irritant (benzalconiumchloride) or solvents did not result in increased signal strength. To determine the cellular origin of IL-18 mRNA in EC, depletion experiments were performed. IL-18 signals were not affected by depletion with anti-CD3 (T cells) or anti-MHC class II mAb-coupled beads identifying KC as a major source of IL-18. These results were confirmed by analysis of mRNA derived from the KC cell line PAM 212. Strong IL-18 signals could be detected by reverse transcription-PCR. To delineate whether IL-18 protein was produced by EC/KC, a sandwich ELISA was used to assay for IL-18 production. Supernatants from allergen-stimulated EC and KC showed production of IL-18 protein. To confirm that IL-18 protein was functional, EC or KC supernatants were tested for their ability to induce IFN-gamma production. Significant amounts of IFN-gamma were detected in supernatants of allergen treated cells. In aggregate, our data indicate that murine KC are a source of both IL-18 mRNA and functional protein. PMID- 9200467 TI - The intronic region of an incompletely spliced gp100 gene transcript encodes an epitope recognized by melanoma-reactive tumor-infiltrating lymphocytes. AB - Recent studies have characterized a number of the Ags that are recognized by melanoma-reactive T cells. Although the majority of tumor Ags appear to represent nonmutated gene products, a variety of epitopes have been shown to arise from either mutated or alternatively processed transcripts. Here, we report that the screening of a cDNA library with a HLA-A24-restricted melanoma-reactive T cell cloid derived from tumor infiltrating lymphocytes resulted in the isolation of a variant of the gp100 gene that had retained the entire fourth intron of this gene, termed gp100-in4. The gp100-in4 transcript could be detected by reverse transcriptase-PCR but could not be detected in Northern blots conducted with melanoma RNA, indicating that it represents a relatively rare transcript. Read through of this transcript into the region corresponding to the fourth intron gave rise to an additional 35 amino acids not found in the normal gp100 glycoprotein, and a peptide within this region conforming to the HLA-A24 consensus motif (VYFFLPDHL) was shown to be recognized by the T cell cloid. The sequence of the intron was identical with that of a previously isolated genomic gp100 clone, and T cells that recognized the gp100-in4 gene product were found to recognize HLA-A24-matched allogeneic melanoma cell lines and melanocytes, demonstrating that this represents a nonmutated epitope. These results further extend the types of Ags that can be recognized by melanoma-reactive T cells to aberrant transcripts of melanosomal genes. PMID- 9200468 TI - NKR-P1A stimulation of arachidonate-generating enzymes in rat NK cells is associated with granule release and cytotoxic activity. AB - NKR-P1A protein has been implicated in the triggering of NK-mediated natural killing contributing to target cell recognition by NK cells. The aim of the present work was to assess whether NKR-P1A receptor triggering also induced arachidonic acid (AA) generation and to determine the possible role of this event on granule release and cytotoxicity. We demonstrated that activation of fresh peripheral blood rat NK cells by cross-linking with the anti-NKR-P1A 3.2.3 mAb induced calcium-dependent AA release, which is due to the activation of cytosolic phospholipase A2 (cPLA2), secretory phospholipase A2 (sPLA2), and diacylglycerol/monoacylglycerol lipase. We also documented the presence of a type II sPLA2 activity in the supernatant fluids from NKR-P1A-activated rat NK cells, suggesting that AA and lysophospholipids could be mobilized from the outside of the cell. The involvement of AA-generating enzymes in NKR-P1A-induced cytotoxic functions was also investigated. Treatment of effector cells with arachidonyl trifluoromethylketone, a cPLA2 inhibitor; p-bromophenacylbromide, a sPLA2 inhibitor; or RHC80267, a diacylglycerol lipase inhibitor, led to a partial inhibition of the redirected lysis against P815 target cells as well the granule content release induced by NKR-P1A cross-linking. A complete abolishment of these events was observed when the cells were simultaneously incubated with all three inhibitors. Taken together, our results support a crucial role for the arachidonate-generating enzymes in the induction of lytic activity of NK cells directly or by leading to generation of additional mediators that can play a role in the context of NK cell activation and cytotoxic functions. PMID- 9200469 TI - Macrophage inflammatory protein-1alpha (MIP-1alpha) is required for the efferent phase of pulmonary cell-mediated immunity to a Cryptococcus neoformans infection. AB - Our objective was to define the role of the chemokine macrophage-inflammatory protein-1alpha (MIP-1alpha) in the efferent phase of pulmonary cell-mediated immunity (CMI) against Cryptococcus neoformans. Following intratracheal inoculation of C. neoformans (24067) into CBA/J mice, the development of CMI was required for leukocyte recruitment into the lungs at 2 wk postinfection. MIP 1alpha mRNA was expressed by day 6 postinfection, and MIP-1alpha protein in bronchoalveolar lavage fluid was detectable at day 6, but significantly elevated at days 19 and 33. Administration of neutralizing anti-MIP-1alpha Abs from days 7 to 13 blocked the increase in bronchoalveolar lavage fluid MIP-1alpha and resulted in a 37% decrease in total leukocytes in the lungs at day 16. There were 66% fewer macrophages/monocytes and 42% fewer neutrophils in the lungs of anti MIP-1alpha-treated mice, and the pulmonary burden of C. neoformans was threefold higher. There was no significant difference in the number of eosinophils, CD4+, CD8+, or B220+ lymphocytes between the two groups of mice. Neutralization of MIP 1alpha did not significantly decrease the levels of monocyte chemotactic protein 1 (MCP-1); however, neutralization of MCP-1 significantly decreased MIP-1alpha levels, demonstrating that induction of MIP-1alpha was largely dependent on MCP-1 production. Neutralization of MIP-1alpha also blocked the cellular recruitment phase of a recall response to cryptococcal Ag in the lungs of immunized mice. Thus, in both the contexts of active cryptococcal infection or rechallenge with cryptococcal Ag, MIP-1alpha was required during the efferent phase of CMI for maximal leukocyte recruitment into the lungs, most notably the recruitment of phagocytic effector cells (neutrophils and macrophages). PMID- 9200470 TI - IL-12 treatment attenuates T helper cell type 2 and B cell responses but does not improve vaccine-enhanced lung illness. AB - In humans and mice, sensitization to respiratory syncytial virus (RSV) Ags can result in severe inflammatory lung disease during subsequent infection with RSV. Although specific antiviral T cells are thought to be responsible for this augmentation of disease, the precise roles of different functional subsets are unknown, and no protective nonpathogenic subset has been defined. BALB/c mice sensitized to the major surface glycoprotein of RSV (G) expressed by recombinant vaccinia virus develop Th2-driven lung eosinophilia after intranasal challenge with RSV. In an attempt to manipulate the outcome of vaccination, we treated mice with IL-12 at various times during vaccination and challenge. IL-12 treatment reduced the vaccine-induced lung eosinophilia during RSV challenge, but increased the total lymphoid cell infiltration into the alveolar space. Analysis of intracellular cytokines by flow cytometry showed that IFN-gamma production during challenge was increased, and IL-4 and IL-5 levels were reduced by IL-12 treatment. In control treated mice, 40 to 50% of the lung lymphoid cells were B cells. Treatment with IL-12 reduced this figure to approximately 1.5%. Although IL-12 treatment reduced lung eosinophilia, illness (as assessed by weight loss) was not eliminated and, in some experiments, was increased. The present study shows that reversing Th2-associated pathology with IL-12 does not necessarily benefit the host. PMID- 9200471 TI - Determinants of T cell reactivity to the Mycobacterium leprae GroES homologue. AB - The 10-kDa protein Ag of Mycobacterium leprae, a human GroES hsp10 cognate, is a major T cell Ag in human leprosy infection. We investigated the mechanism for T cell responsiveness to this Ag according to the trimolecular interaction between T cell, peptide, and Ag-presenting element. This research was accomplished by mapping T cell epitopes in leprosy patients and correlating these responses with peptide-MHC binding affinities. We found that the majority of tuberculoid leprosy patients responded to peptides corresponding to residues 25-39 and 28-42. Truncation analysis of these peptides mapped the exact epitope to be within the overlapping region comprising residues 28-39. Responsiveness was correlated with the HLA-DRB5*0101 allele, which bound the peptides with moderate affinity. This allele is linked to HLA-DR2, which is associated with the resistant form of leprosy. Therefore, T cell responsiveness in tuberculoid leprosy may be mediated by the ability of HLA-DRB5*0101 to bind and present peptides of the immunodominant 10-kDa Ag. PMID- 9200472 TI - Differential CD45 isoform expression accompanies reduced natural antibody binding in L5178Y-F9 tumor progression. AB - Considerable evidence supports a role for polyclonal serum natural Ab (NAb) as a mediator of natural resistance against tumors. However, the molecular mechanisms of this NAb activity are not known. Flow cytometry selection of L5178Y-F9 murine T lymphoma cells for high NAb binding provided the variant LYNAb+, which exhibited an inversely corresponding reduction in tumorigenicity. Accompanying the increased NAb binding, LYNAb+ bound more monoclonal 14.8 anti-CD45RA and DNL 1.9 anti-CD45RC and less 13/2 anti-pan CD45, and the binding of MB23G2 anti CD45RB was eliminated. However, neuraminidase treatment increased NAb binding and detection of pan CD45, CD45RA, and CD45RC but reduced CD45RB expression, suggesting that the epitopes recognized by the former Abs are masked by sialic acid, while the latter includes sialic acid. Growth of the LYNAb+ from a threshold s.c. inoculum in syngeneic DBA/2 mice yielded more tumorigenic cells which bound less NAb, anti-CD45RA, and anti-CD45RC; the same very low anti CD45RB; and more anti-pan CD45. In accord with the mAb binding, the L5178Y-F9 and an in vivo passaged LYNAb+ variant expressed predominantly lower m.w. CD45 isoforms while the LYNAb+ expressed predominantly higher 200-kDa isoforms. The consistent correspondence between CD45RA and CD45RC determinant expression, CD45 isoform expression, tumorigenicity, and NAb binding exhibited by T lymphoma cells selected for high NAb binding in vitro or through tumor progression in vivo suggests that asialo high m.w. isoforms of the cell surface-signaling molecule CD45 are differentially expressed during tumor development and furthermore that they participate in NAb-mediated antitumor mechanisms. PMID- 9200473 TI - Eradication of murine bladder carcinoma by intratumor injection of a bicistronic adenoviral vector carrying cDNAs for the IL-12 heterodimer and its inhibition by the IL-12 p40 subunit homodimer. AB - IL-12 is a heterodimeric immunoregulatory cytokine composed of covalently linked p40 and p35 subunits and exhibits antitumor activity in a variety of laboratory models. The efficacy of systemically administered cytokines for cancer therapy is often limited by toxicity. The gene therapy approach provides a mechanism to achieve temporary and high local concentrations of cytokines within a tumor with less risk of systemic toxicity. We constructed replication-defective adenoviruses containing the murine IL-12 p40 subunit (Ad.mp40) or a bicistronic vector containing cDNAs for the p40 and p35 subunits (Ad.mIL-12). Murine MB49 bladder cancer cells infected with Ad.mIL-12 secrete high concentrations of biologically active IL-12, while those infected with Ad.mp40 produce the p40 homodimer. Tumors injected with Ad.mIL-12 show rapid increases in IL-12 and IFN-gamma expression over 2 to 5 days and a return to baseline by 7 to 14 days. Injection of tumors with Ad.mIL-12 (1 x 10(9) plaque-forming units) results in a complete tumor regression in all mice, while those treated with control adenovirus succumb to their tumor. Efficacy is reduced when studies are performed in mice depleted of CD4+ and CD8+ cells or in nude mice. Mice cured of their tumor by Ad.mIL-12 demonstrate specific protective immunity upon rechallenge. Ad.mp40 does not exhibit antitumor activity and may antagonize the activity of rIL-12 or Ad.mIL 12. In summary, gene therapy strategies for cancer using adenoviral vectors containing IL-12 are highly effective with no significant toxicity in mice. PMID- 9200474 TI - A mechanism for selective recruitment of CD8 T cells into B7-1-transfected plasmacytoma: role of macrophage-inflammatory protein 1alpha. AB - An important aspect of immunity is the recruitment and accumulation of lymphocytes into target tissues where Ags are localized. Because the TCR recognizes antigenic peptides presented by MHC molecules, the subset of T cells that exert effector function is determined by the class of MHC molecules expressed on a given tissue. We and others have demonstrated that CD8 T cells are preferentially recruited into B7-1-transfected class II-negative plasmacytoma J558, and the virus-infected central nervous system. However, the mechanism for such specificity has not been addressed. Here we analyzed the mechanism for selective recruitment of CD8 T cells into B7-1-transfected plasmacytoma J558. We show that sustained accumulation of CD8 T cells in vivo requires local expression of B7-1. In addition, we have observed a striking correlation between expression of macrophage-inflammatory protein 1alpha (MIP1alpha) and selective accumulation of CD8 T cells in the tumors. The selective recruitment of CD8 T cells is blocked by in vivo administration of neutralizing anti-MIP1alpha antisera. Moreover, gene transfer studies reveal that locally produced MIP1alpha is sufficient to induce selective recruitment of CD8 T cells. Taken together, our study reveals that costimulation by B7 leads to sustained local production of MIP1alpha, which selectively recruits CD8 T cells into tumors. These results have important implications for T cell recruitment in vivo and for tumor immunotherapy. PMID- 9200475 TI - Expression of a biologically active recombinant mouse IL-1 receptor antagonist and its use in vivo to modulate aspects of the acute phase response. AB - Recombinant mouse IL-1receptor antagonist protein (rmIL-1ra) was expressed in Escherichia coli. In vivo administration of rmIL-1ra, in a casein-induced murine model of acute inflammation, completely abolished the hepatic induction of the mRNAs specifying serum amyloid A1 (A-SAA1) and A-SAA2 for up to 12 h, indicating that hepatic A-SAA mRNA synthesis is totally IL-1 driven. A-SAA protein, however, was present in the serum of rmIL-1ra-treated casein-stimulated mice (although at lower levels than in untreated casein-stimulated mice) at 12 h indicating that extrahepatic A-SAA synthesis is driven in part by factors acting independently of IL-1. Hepatic mRNA levels of the other mouse acute phase reactants (APRs), serum amyloid P component, C-reactive protein, alpha1-acid glycoprotein, and C3 were also induced with casein after 12 h, as were serum protein levels of SAP and C3. These inductions were only partially inhibited by rmIL-1ra, indicating that hepatic expression of the latter APRs (unlike that of A-SAA) is driven partly by IL-1 and partly by factors acting independently of IL-1. Hepatic mRNA levels of the negative APRs apolipoprotein A-I and serum albumin were down-regulated 12 h after casein stimulation. rmIL-1ra partially restored serum albumin mRNA levels but not apo A-I mRNA levels, indicating differential regulation of these negative APRs. The rmIL-1ra will be useful in studies of IL-1-mediated gene regulation in murine models of inflammation. PMID- 9200476 TI - IL-13 inhibits TNF production but potentiates that of IL-6 in vivo and ex vivo in mice. AB - IL-13 was reported to inhibit the synthesis of various cytokines in vitro, including that of TNF. It has divergent effects on IL-6 production, which is increased in endothelial cells and decreased in monocytes. We studied the effect of IL-13 administration on TNF and IL-6 production in vivo in mice. IL-13 (1 microg/mouse, i.v., 10 min to 6 h before LPS) decreased LPS (100 ng/mouse, i.v.) induced serum TNF levels by 50%, while it increased the levels of IL-6 by fourfold. IL-13 potentiated IL-1beta (100 ng/mouse, i.v.)-induced serum IL-6 levels as well as IL-1- or LPS-induced serum amyloid A. When blood from IL-13 treated mice was stimulated with LPS in vitro, TNF production was decreased fivefold, and that of IL-6 was slightly decreased. We also cultured in vitro the aorta obtained from IL-13-pretreated mice and found that they produce more IL-6 (up to sevenfold) than aorta from control mice. Little or no TNF could be detected in these samples. Thus, IL-13 in vivo inhibits serum TNF but up regulates serum IL-6. The differential regulation of IL-6 and TNF together with the results of ex vivo experiments could be explained by hypothesizing that the cellular origins of the two cytokines are different. PMID- 9200477 TI - Effect of the inducible nitric oxide synthase inhibitors aminoguanidine and L-N6 (1-iminoethyl)lysine on zymosan-induced plasma extravasation in rat skin. AB - The effect of nitric oxide synthase (NOS) inhibitors on plasma extravasation in a rat model of zymosan-induced inflammation has been investigated. Plasma extravasation was determined in response to intradermal test agents over 0 to 45 min or 0 to 4 h by the accumulation of i.v. injected 125I-labeled human serum albumin. Zymosan (1-100 microg/site) produced a dose- and time-dependent plasma extravasation. N(G)-nitro-L-arginine methyl ester (30-300 nmol/site), but not aminoguanidine (AG; 10-300 nmol/site) or L-N6-(1-iminoethyl)lysine (L-NIL; 10-300 nmol/site), significantly (p < 0.01) inhibited zymosan-induced (10 microg/site) plasma extravasation over 0 to 45 min. However, both AG and L-NIL produced significant (p < 0.05) inhibition over 0 to 4 h. The inhibition produced by AG was reversed by i.v. L-arginine or by coinjection of the vasodilator, calcitonin gene-related peptide. Zymosan (10-100 microg/site) induced an increase in dermal blood flow (laser-Doppler flowmetry) and this was inhibited by AG. Neutrophils were depleted selectively with antiserum, but this did not affect plasma extravasation except at the highest dose of zymosan (100 microg/site). Furthermore, zymosan-induced edema was not modified at either time point by pretreatment with the cyclooxygenase inhibitor indomethacin (30 micromol/kg, s.c., -30 min). In conclusion, in this model of dermal inflammation, it is suggested that inducible NOS inhibitors selectively remove an inducible NOS component that, at least in part, acts to increase microvascular blood flow and thus the edema formation observed during 0 to 4 h. There is no evidence of a contributory role for neutrophils or cyclooxygenase products in this model. PMID- 9200478 TI - Endotoxin induces expression of type II phospholipase A2 in macrophages during acute lung injury in guinea pigs: involvement of TNF-alpha in lipopolysaccharide induced type II phospholipase A2 synthesis. AB - Elevated levels of secretory type II phospholipase A2 (sPLA2-II) have been associated with a poor clinical outcome in the acute respiratory distress syndrome. This study identifies the cell source(s) and the mechanisms of sPLA2-II synthesis in the guinea pig model of acute respiratory distress syndrome induced by intratracheal injection of LPS. Administration of LPS led to an increase in lung membrane-associated calcium-dependent sPLA2 activity, which was abrogated by LY311727, a selective inhibitor of sPLA2-II. No sPLA2 activity was detected in the vascular compartment of the lung. LPS administration induced a parallel accumulation of sPLA2-II mRNA in lung tissues. In situ hybridization showed that sPLA2-II transcripts were synthesized in interstitial and alveolar macrophages (AM). Incubation of AM with LPS enhanced the expression of sPLA2-II mRNA, leading to stimulation of sPLA2-II synthesis and secretion. This increase was prevented by the addition of anti-TNF-alpha and anti-p55 TNF receptor Abs. Furthermore, the addition to AM of cellfree bronchoalveolar fluid collected from LPS-treated guinea pigs increased sPLA2-II expression, which was abrogated by anti-TNF-alpha Ab. These findings demonstrate that 1) macrophages are in vivo the major cell source of sPLA2-II in LPS-induced acute lung injury; 2) in contrast to that in other cell systems, regulation of LPS-induced sPLA2-II synthesis in AM is TNF alpha dependent; and 3) production of TNF-alpha in the air-lung interface is an important step for sPLA2-II synthesis in macrophages. PMID- 9200480 TI - Fc receptor-triggered insertion of secretory granules into the plasma membrane of human neutrophils: selective retrieval during phagocytosis. AB - We studied the kinetics of secretion in human neutrophils stimulated by IgG opsonized zymosan. Secretion of azurophilic and specific granules was quantified measuring the appearance of the granule markers CD63 and CD66b, respectively, at the cell surface. The kinetics of secretion was compared with the course of phagocytosis, revealed by the trapping of the fluid phase marker, Lucifer Yellow, in vacuoles containing zymosan particles. We found that secretion of both azurophilic and specific granules precedes phagosome sealing. An initial rapid phase of secretion was followed by a decrease in the amount of CD63 and CD66b at the cell surface. This subsequent disappearance of surface CD63 and CD66b was inhibited by cytochalasin B and probably represents internalization of the granular markers into the forming phagosome. The decrease in the amount of CD63 and CD66b exposed at the cell surface was not accompanied by a commensurate reduction in cell surface area, measured with the amphiphilic fluorescent dye FM1 43. These findings imply that CD63 and CD66b are selectively retrieved from the plasma membrane following secretion. Evidence is also presented that calcium is not the sole mediator of the rapid secretion of azurophilic and specific granules triggered by IgG-opsonized particles and that cytochalasin does not impair signaling of the calcium transient elicited by Fc receptors. Instead, actin disassembly appears to reduce the efficiency of the interaction between opsonized particles and their receptors, an effect that can be overcome by increasing the concentration of the stimulating particles. PMID- 9200479 TI - Transgenic monocyte chemoattractant protein-1 (MCP-1) in pancreatic islets produces monocyte-rich insulitis without diabetes: abrogation by a second transgene expressing systemic MCP-1. AB - Monocyte chemoattractant protein-1 (MCP-1) is a CC chemokine that attracts monocytes and T lymphocytes in vitro; however, its in vivo functions are poorly understood. To address this question, we constructed transgenic mice expressing MCP-1 controlled by an insulin promoter. These mice developed a chronic insulitic infiltrate composed of F4/80+ monocytes with minor populations of CD4+, CD8+, and B220+ cells. Despite persistent transgene expression, the insulitis never progressed, and blood glucose levels remained normal. Thus, MCP-1 alone is sufficient to elicit a monocytic infiltrate, but not to activate elicited cells. These results differ from those obtained with another transgenic model using the mouse mammary tumor virus long terminal repeat, in which mice expressed substantial MCP-1 in several organs but had no infiltrates. However, mice expressing both transgenes had minimal insulitis, indicating that high systemic levels of MCP-1 prevented monocytes from responding to local MCP-1. Thus, the ability of MCP-1 to elicit monocytic infiltration depends on its being expressed at low levels in an anatomically restricted area. PMID- 9200481 TI - Differential regulation of indoleamine 2,3-dioxygenase expression by nitric oxide and inflammatory mediators in IFN-gamma-activated murine macrophages and microglial cells. AB - Induction of indoleamine 2,3-dioxygenase (IDO) and nitric oxide synthase (NOS) is involved in the immunomodulatory roles of IFN-gamma and evidence suggests that these pathways are functionally cross-regulated. We report here that nitric oxide (NO) negatively modulates the expression of IDO activity in IFN-gamma-primed macrophages, but not in microglial cells from mouse. In MT2 macrophages, the induction of IDO activity by IFN-gamma was further increased by the presence of NOS inhibitors, whereas culturing of IFN-gamma-activated MT2 cells with NO generators produced a marked reduction of IDO activity expression. Conversely, neither NOS inhibitors nor exogenous NO affected the induction of the enzyme activity in N11 microglial cells after IFN-gamma activation. LPS and picolinic acid, two costimulatory agents that up-regulate inducible NOS in activated cells, regulated IDO induction differently in the two cell lines. LPS and picolinic acid caused a significant decrease of IDO activity in IFN-gamma-activated MT2 cells. This effect, however, did not appear to be mediated by the ability of LPS and picolinic acid to stimulate NO production. In N11 cells, LPS further stimulated the enzyme activity and picolinic acid had no effect. Northern blot analysis revealed that, in MT2 macrophages, NOS inhibitors increased the levels of IDO mRNA, while a reduction was observed with picolinic acid. No changes in IDO mRNA levels were detected in N11 cells. Consistent with the functional heterogeneity of phagocytes, the reported results indicate the existence of marked differences in the regulation of IDO expression between murine macrophages and microglial cells. PMID- 9200482 TI - Shared TCR Vbeta gene expression by the pancreas and salivary gland in immunodeficient alymphoplasic mice. AB - Mice with the homozygous mutation alymphoplasia (aly) lack lymph nodes and Peyer's patches and show defects in both humoral and cellular immunity. In these mice, spontaneous infiltration of mononuclear cells was observed in multiple exocrine organs, including the pancreas, salivary glands, and lacrimal glands from the age of 15 wk, progressing to a marked tissue destruction at the age of 25 wk. Using this strain, we examined the phenotypes and TCR Vbeta gene expression of infiltrating T cells to identify the pathologic role of T cell immunity in idiopathic pancreatitis. Most of the infiltrating cells were CD4+ and Thy-1+ cells. Analysis of the TCR gene expression on T cells infiltrating the pancreas and salivary glands showed a high expression of Vbeta1 and Vbeta5 in both organs at the age of 15 wk. In contrast, a diverse expression of TCR Vbeta genes was noted at 25 wk. Sequence analysis of complementarity-determining region 3 (CDR3) of the most prominent TCR Vbeta gene family expressed in these cells, Vbeta1, showed oligoclonal expansion of infiltrating T cells in both organs. Frequent use of glutamine and proline at position 97 was observed in paired tissues. Our data suggest that oligoclonal expansion of organ specific T cells might be one of the etiologic mechanisms of chronic pancreatitis and that common autoantigens could trigger autoimmunity in multiple organs. PMID- 9200483 TI - Outside-in signaling by lipopolysaccharide through a tailless integrin. AB - Ligand binding to integrins activates intracellular signaling pathways that coordinate and regulate a variety of cellular responses. There is evidence to suggest that the cytoplasmic tails play a key role in several of these signaling events. We sought to determine whether the beta2 integrin complement receptor type 3 (CR3; CD11b/CD18), a receptor for LPS, could initiate an intracellular signal in the absence of its cytoplasmic domains. Expression of full length CR3 in a Chinese hamster ovary-K1 fibroblast line enabled serum-independent translocation of nuclear factor-kappaB in response to binding LPS. Unexpectedly, a cell line expressing a mutated form of CR3 deficient in the cytoplasmic domains was also competent for transmitting a signal in response to LPS. In contrast, phagocytosis of whole Gram-negative bacteria and iC3b-coated erythrocytes took place only with a full length receptor. Thus, while full length CR3 is necessary for productive phagocytic signals, LPS activation does not require the cytoplasmic domains. CR3 may function to activate cells by presenting LPS to a downstream signal transducer. PMID- 9200484 TI - Cyclooxygenase-2-dependent delayed prostaglandin D2 generation is initiated by nerve growth factor in rat peritoneal mast cells: its augmentation by extracellular type II secretory phospholipase A2. AB - When rat serosal connective tissue mast cells (CTMC) were stimulated with nerve growth factor (NGF), the immediate prostaglandin D2 (PGD2) generation was followed by delayed PGD2 generation that occurred between 2 and 24 h, reaching levels as high as 50 ng and 260 ng/10(6) cells in the absence or presence of lysophosphatidylserine (lysoPS), respectively. This delayed PGD2 generation was accompanied by de novo induction of cyclooxygenase (COX)-2, with NGF and lysoPS acting as inducer and enhancer, respectively. COX-2 induction and the attendant delayed PGD2 generation in CTMC were modestly induced by c-kit ligand, but not by Fc epsilonRI cross-linking. This indicated that the stimulus specificity differed from that observed in the immediate phase, in which NGF, c-kit ligand, and Fc epsilonRI cross-linking, either in combination with each other or with lysoPS as a cofactor, elicited comparable levels of PGD2 generation within 10 min, reaching 10 to 20 ng/10(6) cells. Addition of type II secretory phospholipase A2 (sPLA2), a PLA2 isoform that is detected in microg/ml levels in inflammatory exudates, to NGF-stimulated CTMC significantly augmented delayed, but not immediate, PGD2 generation, and this augmentative effect was mediated in part by the enhancement of COX-2 expression by sPLA2. These results suggest that CTMC have the capacity to produce PGD2 over a prolonged period in the presence of tissue-derived cytokines and sPLA2 in a COX-2-dependent manner. PMID- 9200485 TI - Recombinant histamine-releasing factor enhances IgE-dependent IL-4 and IL-13 secretion by human basophils. AB - Human recombinant histamine-releasing factor (HrHRF) is known to directly stimulate histamine release and IL-4 secretion from basophils of selected atopic donors in a reaction requiring the expression of a particular type of IgE, referred to as IgE+. In this study, HrHRF is shown to affect the IgE-mediated release of IL-4, IL-13, and histamine from basophils not normally releasing to this protein (i.e, those expressing IgE-). Priming with several different concentrations of HrHRF for 15 min enhanced basophil secretion of IL-4 and histamine after 4 h in a dose-dependent fashion following activation with anti IgE Ab (10 ng/ml). This effect of HrHRF priming also occurred in cultures activated with 1 or 100 ng/ml of anti-IgE Ab. The secretion of IL-13 protein was enhanced similarly by HrHRF priming in cultures stimulated for 16 to 20 h with anti-IgE Ab. There were, however, no apparent changes in the secretion of histamine or cytokine by basophils primed with HrHRF and activated with the IgE independent secretogogue, FMLP. These findings suggest that HrHRF modifies the response of basophils for IgE-dependent secretion by binding to a specific receptor, broadening the possible role of this protein in chronic allergic inflammation. PMID- 9200486 TI - Genetic dissection of systemic lupus erythematosus pathogenesis: Sle2 on murine chromosome 4 leads to B cell hyperactivity. AB - Susceptibility to systemic lupus erythematosus in the NZM2410 murine model maps to Sle1, Sle2, Sle3, and the H2 loci. To unravel how these loci contribute to the pathogenesis of lupus, individual NZM2410-derived genomic intervals bearing these loci have been successfully backcrossed onto the resistant C57BL/6 (B6) background. The focus of this study was to understand how Sle2 on murine chromosome 4 impacts the immune system. Compared with C57BL/6 (B6) mice, B6 mice congenic for Sle2 exhibit a variety of immunophenotypes affecting their B cells. They have an early, but transient, expansion of splenic, CD23(low) B cells. Thereafter, their B cells appear activated by surface phenotype and functional criteria, paralleled by elevated serum levels of polyreactive/polyclonal IgM. Importantly, Sle2 leads to a heightened B cell responsiveness to in vitro stimuli and to in vivo antigenic challenge. Finally, they exhibit increased levels of peritoneal and splenic B1 cells. Thus, Sle2 harbors a gene that leads to B cell hyperactivity and elevated B1 cell formation. However, Sle2 by itself on the normal B6 background is insufficient to generate IgG antinuclear Abs (ANA) or nephritis. By reducing the B cell signaling threshold, Sle2 might serve to amplify an ongoing autoimmune response. PMID- 9200487 TI - Role of macrophages and macrophage-derived cytokines in the pathogenesis of Kilham rat virus-induced autoimmune diabetes in diabetes-resistant BioBreeding rats. AB - The diabetes-resistant BioBreeding (DR-BB) rat, derived from diabetes-prone forebears, does not normally develop spontaneous insulitis or diabetes, but when infected with Kilham rat virus (KRV) this animal develops autoimmune diabetes similar to the diabetes-prone BioBreeding (DP-BB) rat. In this study, we attempted to determine whether macrophages and macrophage-derived cytokines play a role in the development of KRV-induced diabetes in DR-BB rats. Seventy-eight percent of DR-BB rats treated with KRV and poly(I:C) develop diabetes, whereas depletion of macrophages with liposome-encapsulated dichloromethylene diphosphonate (lip-Cl2MDP) in KRV and poly(I:C)-treated DR-BB rats results in the near-complete prevention of insulitis and diabetes. Measurement of the macrophage derived cytokines IL-12, TNF-alpha, and IL-1beta revealed a selective increase of their expression, after KRV infection, in the splenic lymphocytes and the pancreatic islets. Measurement of CD4+ T cell-derived cytokines revealed that IL 2 and IFN-gamma cytokine gene expression closely correlates with an elevation of IL-12, but IL-4 and IL-10 do not change. Depletion of macrophages before the isolation of splenic lymphocytes from DR-BB rats treated with KRV and poly(I:C) resulted in the loss of ability to transfer diabetes to young DP-BB rats. On the basis of these observations, we conclude that macrophages and macrophage-derived cytokines play a critical role in the cascade of events leading to the destruction of pancreatic beta cells, culminating in the development of insulin dependent diabetes mellitus. PMID- 9200488 TI - Identification of rat prostatic steroid-binding protein as a target antigen of experimental autoimmune prostatitis: implications for prostate cancer therapy. AB - The long term goal of this study is to develop autoimmune prostatitis as a therapy for prostate cancer. An immune attack capable of destroying normal prostate epithelial cells should also destroy malignant prostate tissue and provide therapeutic benefit in cancer patients. The current study was initiated to identify antigenic targets for experimental autoimmune prostatitis on the assumption that such proteins might also be suitable targets for immunotherapy of prostate cancer. Male Lewis rats were immunized with syngeneic prostate homogenates, and the immune sera were used to screen prostate proteins for immunoreactivity by Western blot analysis. The dominant protein recognized by the immune sera was purified by ion exchange chromatography and reverse phase HPLC. Microsequence analysis of two polypeptide components of this immunodominant protein demonstrated N-terminal sequences identical with two of the three component chains of rat prostatic steroid-binding protein (PSBP). T cell responses to PSBP were also detected in rats immunized with prostate homogenate. Immunizing male rats with purified PSBP induced vigorous Ab and T cell responses. Significant prostate inflammation was observed in some rats immunized with PSBP. Adoptive transfer of T cells immune to PSBP induced rapid and severe destructive autoimmune prostatitis. These results demonstrate that PSBP is a major target Ag of experimental autoimmune prostatitis in a rat model and may serve as a target Ag for vaccine and T cell therapy against prostate cancer. PMID- 9200489 TI - Clonal expansion of CD8+ T cells in Kawasaki disease. AB - Kawasaki disease (KD) is the major cause of acquired heart disease in children. KD is suspected of being an infectious disease, but the etiology has not yet been clarified. Immunologically, the disease is associated with the activation of T cells, monocytes, and macrophages resulting in highly elevated levels of several cytokines. Recently, expansions of T cells expressing TCRBV2 and TCRBV8 chains have been reported, and this suggests the involvement of a superantigen in the pathogenesis of KD. To address the role of a superantigen in KD, we investigated clonal expansion of T cells by estimating the complementarity-determining region 3 size profile among T cells expressing TCRBV1, TCRBV2, TCRBV4, TCRBV5, TCRBV8, TCRBV14, TCRBV16, TCRBV17, TCRBV18, and TCRBV20 chains during acute KD, during subacute KD, and during the long term follow-up period. During the acute phase of KD, several clonal expansions were found mainly in the CD8+ T cells that disappeared during the long term follow-up period. Our data suggest that the conventional Ags rather than a superantigen were involved in the pathogenesis of acute KD. PMID- 9200490 TI - A role for insulin-like growth factor in the regulation of IL-6-responsive human myeloma cell line growth. AB - Insulin-like growth factors (IGF-I, IGF-II) have long been recognized as important mitogens in many types of malignancies. Because the role of IGFs in growth control of myeloma cells has not been extensively examined, we have used a panel of IL-6-responsive myeloma cell lines to address this issue. Initial studies demonstrated that IGF-I and IGF-II significantly enhanced DNA synthesis by each of the four cell lines, even when assayed in the absence of IL-6. The specificity of the IGF response was confirmed using an IGF-I receptor Ab, and additional studies demonstrated that IGF responsiveness did not result from induction of autocrine IL-6 expression. When IL-6 responsiveness was assayed, three of four cell lines synthesized DNA in response to IL-6 alone; however, the magnitude of responsiveness was greatly enhanced by addition of IGFs. Similar results were obtained when proliferation and cell cycle progression were analyzed. By contrast, the KP-6 cell line was responsive to IL-6 only when IGF was present. Finally, we analyzed the effects of IGF-I on normal B lymphocytes. IGF, however, did not stimulate B cell DNA synthesis, suggesting that IGF responsiveness may represent a key difference between normal and malignant B cells. In summary, these studies suggest that IGFs may play an important role in multiple myeloma by virtue of their ability to directly stimulate tumor cell growth as well as modulate the magnitude of IL-6-driven growth. PMID- 9200491 TI - Triggers of autoimmune disease in a murine TCR-transgenic model for multiple sclerosis. AB - The combination of genetic and environmental factors that contribute to human autoimmune responses has made potential triggers of these diseases difficult to identify. We examined how experimental allergic encephalomyelitis (EAE), an animal model of multiple sclerosis, is triggered using TCR-transgenic mice specific for myelin basic protein (MBP). In these TCR-transgenic mice, EAE can be actively induced and also occurs spontaneously. The incidence of spontaneous EAE in this model is largely confined to adolescence and early adulthood and is more prevalent among males than females, indicating that hormonal influences may contribute to triggering central nervous system autoimmune disease. Disease induction studies show that not all stimuli that activate MBP-specific T cells in vivo also induce EAE. Immunization with MBP peptide stimulates the transgenic T cells to produce Th1 cytokines; however, the activated T cells do not accumulate in the central nervous system and induce EAE unless pertussis toxin is also administered. EAE can be induced by intrathecal injection of either stimulated or nonstimulated transgenic T cells into nontransgenic or transgenic recipients. Therefore, gaining access to the central nervous system appears to be the critical step in this model for the induction of EAE, regardless of the activation state of the T cells. PMID- 9200492 TI - Subarachnoid hemorrhage impairs cerebral blood flow response to nitric oxide but not to cyclic GMP in large cerebral arteries. AB - Nitric oxide (NO) increases 3',5'-cyclic guanosine monophosphate (cGMP) in vascular smooth muscle and increases cerebral blood flow (CBF). In early stages of cerebral ischemia, NO plays a beneficial role in sustaining CBF. Subarachnoid hemorrhage (SAH), one of the main causes of ischemia, may impair vascular reactivity to NO. To test the hypothesis, 48 h after SAH was induced in rats, we examined the CBF response to the NO donor, SIN-1 (3-morpholinosydnonimine). We measured CBF by laser-Doppler flowmetry in association with: (1) intracarotid injection (for 30 min) of SIN-1 (1.5 mg/kg), 8-bromo-cGMP (7.5 mg/kg), papaverin (1.5 mg/kg) or vehicle; (2) cortical superfusion (for 90 min) of SIN-1 (10(-5) M) or vehicle through the cranial window. Hypotension produced by these vasodilators was controlled with phenylephrine. Vehicle alone did not change CBF throughout the measurement. Intracarotid infusion of SIN-1 (n = 6/group) increased CBF up to 128.6 +/- 3.9% and 111.9 +/- 2.9% in the control group and the SAH group, respectively. SAH significantly attenuated the response (P < 0.05, ANOVA). SAH did not affect the CBF increases elicited by intracarotid administration of cGMP or papaverin, or cortical superfusion of SIN-1. We conclude that during chronic vasospasm SAH disturbs the pathway between NO release and cGMP production in large cerebral arteries. The impairment accounts for the fragility of the brain in the face of ischemia following SAH. PMID- 9200493 TI - Effects of estrogen on basal forebrain cholinergic neurons vary as a function of dose and duration of treatment. AB - Studies suggest that estrogen replacement can influence learning and memory processes via effects on cholinergic neurons located in specific regions of the basal forebrain. In the present study, immunocytochemical techniques were used to examine the effects of estrogen on basal forebrain cholinergic neurons as a function of the dose and duration of estrogen treatment. Ovariectomized rats received 2, 10, 25, or 100 microg estradiol every other day for a period of 1, 2, or 4 weeks. Sections through the basal forebrain were then processed for the detection of choline acetyltransferase (ChAT) or the low-affinity nerve growth factor receptor (p75NGFR), and the number of immunoreactive cells in the medial septum (MS), the horizontal limb of the diagonal band of Broca (HDB) and the nucleus basalis magnocellularis (NBM) were counted. The effects of dose and duration of estrogen treatment were evaluated by analysis of variance and individual group means were compared with ovariectomized controls using a two tailed Dunnets test. Administration of 2, 10, or 25 microg estradiol for 1 week produced a dose-related increase in the number of ChAT-like immunoreactive (IR) cells detected in the MS. Likewise treatment with 10 microg estradiol for 1 week, or with 2 microg estradiol for 2 weeks resulted in a significant increase in the number of ChAT-IR cells detected in the NBM. These effects were not observed following treatment with higher doses of estradiol. Nor were they maintained following repeated administration of estradiol for longer periods of time. In contrast, repeated administration of estradiol for 2 or 4 weeks resulted in significant decreases in the number of p75NGFR-IR cells detected in the MS, with the greatest effects observed following treatment with the higher doses of estradiol for longer periods of time. These findings demonstrate that (1) estrogen replacement produces regionally selective effects on basal forebrain cholinergic neurons which vary as a function of both the dose and duration of estrogen treatment, and (2) estrogen has both short-term and longer-term effects on basal forebrain cholinergic neurons, each of which may contribute to the effects of estrogen on learning and memory process and the development of age- and disease-related cognitive decline. PMID- 9200494 TI - 5-HT2 receptor regulation of acetylcholine release induced by dopaminergic stimulation in rat striatal slices. AB - The role of 5-hydroxytryptamine (5-HT) receptor subtypes in acetylcholine (ACh) release induced by dopamine or neurokinin receptor stimulation was studied in rat striatal slices. The dopamine D1 receptor agonist SKF 38393 potentiated in a tetrodotoxin-sensitive manner the K(+)-evoked [3H]ACh release while SCH 23390, a dopamine D1 receptor antagonist, had no effect. [3H]ACh release was decreased by the dopamine D2 receptor agonist LY 171555 (quinpirole) and slightly potentiated by the dopamine D2 receptor antagonist haloperidol. The selective neurokinin NK1 receptor agonist [Sar9, met(O2)11]SP also potentiated K(+)-evoked release of [3H]ACh. GR 82334, a NK1 receptor antagonist, blocked not only the effect of [Sar9, met(O2)11]SP but also the release of ACh induced by the D1 receptor agonist SKF 38393. Among the 5-HT agents studied, only the 5-HT2A receptor antagonists ketanserin and ritanserin were able to reduce the ACh release induced by dopamine D1 receptor stimulation. Mesulergine, a more selective 5-HT2C antagonist, showed an intrinsic releasing effect but did not affect K(+)-evoked ACh release induced by SKF 38393. Methysergide and methiothepin, mixed 5-HT1/2 antagonists, as well as ondansetron, a 5-HT3 receptor antagonist, showed an intrinsic effect on ACh release, their effects being additive to that of SKF 38393. 5-HT2 receptor agonists were ineffective. However, the 5-HT2 agonist DOI was able to prevent the antagonism by ketanserin of the increased [3H]ACh efflux elicited by SKF 38393, suggesting a permissive role of 5-HT2A receptors. None of the above indicated 5-HT agents was able to reduce the ACh release induced by the selective NK1 agonist. The results suggest that 5-HT2 receptors, probably of the 5-HT2A subtype, modulate the release of ACh observed in slices from the rat striatum after stimulation of dopamine D1 receptors. It seems that this serotonergic control is exerted on the interposed collaterals of substance P containing neurons which promote ACh efflux through activation of NK1 receptors located on cholinergic interneurons. PMID- 9200495 TI - Insect 86 kDa protein kinase C substrate is a filament interacting protein regulated by Ca2+/calmodulin and phosphorylation. AB - A specific substrate for protein kinase C (PKC) has been purified to apparent homogeneity from neuronal tissue of the honeybee Apis mellifera. The phosphoprotein shows an apparent molecular mass of 86 kDa, generates multiple spots around the pH of 4.6 upon isoelectric focussing and shows phosphopeptide patterns after limited proteolysis that are nearly identical to those of myristoylated alanine-rich C kinase substrate (MARCKS) purified from bovine brain. Investigations of the functional properties show, that the 86 kDa protein is regulated by both, the phosphorylation by PKC and by Ca2+/calmodulin. The phospho- and dephospho-86 kDa protein bind to F-actin with dissociation constants of K(d) approximately 280 nM and K(d) = 690 nM, respectively. Ca2+/calmodulin inhibits both, the phosphorylation of the 86 kDa protein by PKC and its interaction with F-actin. These data strongly suggest that the insect 86 kDa protein is a potential site of convergence of the Ca2+/calmodulin and PKC signalling pathways in the regulation of cytoskeleton-membrane rearrangement, with structural and functional homologies to vertebrate MARCKS. PMID- 9200496 TI - Involvement of nitric oxide in re-innvervation of rat molar tooth pulp following transection of the inferior alveolar nerve. AB - The aim of this study was to elucidate whether nitric oxide (NO) is involved in re-innervation of rat molar tooth pulp following transection of the inferior alveolar nerve. The inferior alveolar nerves (IAN) of rats were transected unilaterally under anesthesia with chloral hydrate. The animals received horseradish peroxidase (HRP) application to mandibular molar tooth pulps on both sides and were fixed by transvascular perfusion. The average number of labeled cells on each side of the trigeminal ganglion was not significantly different [101 +/- 11 (mean +/- S.E.M.; n = 6, left) and 89 +/- 11 (n = 6, right)]. With HRP application on postoperative day 3, the ratio of the number of labeled neurons in the transected vs. non-transected (contralateral) sides was 31.5 +/- 5.8% (n = 11). The i.p. administration of N(omega)-nitro-L-arginine methyl ester (L-NAME; 100 mg/kg, once a day for a period of 4 days), but not D-NAME, significantly decreased the ratio of the number of labeled neurons (10.1 +/- 7.0%, n = 10). L-Arginine (300 mg/kg, i.p., once a day for a period of 4 days) slightly increased the number of labeled neurons on the transected side. Clonidine (25 microg/kg, i.p., once a day for a period of 4 days) failed to exhibit any significant effect on nerve regeneration. In the trigeminal ganglion ipsilateral to the transected IAN on postoperative day 4, NADPH-diaphorase (NADPH d)-positive neurons had significantly increased. On the other hand, no changes in NADPH-d were observed in the superficial layers of the subnucleus caudalis of the spinal trigeminal nucleus from where primary neurons innervating the mammalian tooth pulp project. These results suggest that NO is involved in several mechanisms related to neuronal regeneration. PMID- 9200497 TI - Involvement of spinal cord delta opiate receptors in the antinociception of gestation and its hormonal simulation. AB - Physiological as well as hormone-simulated pregnancy (HSP) is associated with opioid-mediated elevations in maternal nociceptive thresholds. Previous reports from this laboratory have demonstrated the involvement of spinal cord kappa opiate receptors in this phenomenon. The present study was undertaken in order to determine the exclusivity of this mediation. Intrathecal (i.t.) administration of the delta opiate receptor-selective antagonists naltrindole (NTI), 7 benzylidenenaltrexone (BNTX) or naltriben (NTB) substantially reduces nociceptive thresholds of gestation (day 20) and HSP (day 19). Hyperalgesic actions of these compounds following i.t. administration are not observed in non-pregnant or vehicle-treated control animals. These data indicate that delta opiate receptor activity is a prerequisite for the manifestation of a substantial portion of gestational and HSP analgesia. In contrast, i.t. application of the micro selective antagonist D-Phe-Cys-Tyr-D-Trp-Arg-Thr-Pen-Thr-NH2 (CTAP) has no effect on nociceptive thresholds of gestational day 20, as was previously demonstrated for HSP-induced antinociception. Thus, the potent spinal mu analgesic system does not participate in gestational or HSP analgesia. During physiological pregnancy, less robust constituents of intrinsic opioid pain-attenuating systems in the spinal cord (delta and kappa opioid systems) are recruited to mediate the maternal antinociception of gestation. Furthermore, the ability of estrogen and progesterone to modulate spinal opioid antinociceptive activity emphasizes potential differences between men and women in their response to pain medication. PMID- 9200498 TI - Spatial zones for muscle coactivation and the control of postural stability. AB - It is hypothesized that, depending on the motor task, the angular range of a joint may be subdivided into zones in which agonist and antagonist muscles are coactive, only one group of muscles is active or neither group is active. It is further hypothesized that central commands may change the size and location of these spatial zones. We investigated whether spatial zones are used by the nervous system and how they may be changed to provide postural stability of the elbow. We compared responses to sudden unloading of the elbow flexors in neurologically normal subjects with those in patients with postural control deficits due to unilateral hemispheric and/or subcortical lesions. By studying responses in patients, we sought to determine whether the specification of zones of agonist/antagonist muscle coactivation ("coactivation zones") may be essential for postural stability. At an initial elbow angle (130 degrees; full extension is 180 degrees), flexors were pre-activated by compensating an initial load which was equal to approximately 30% of the subject's maximal isometric voluntary contraction effort. Subjects were instructed not to correct the arm displacement elicited by a sudden decrease in the load. Data from 10 trials were collected at each of 4-6 final load levels (separated by 1.5-2 Nm) in order to map out the relationship between torque and angle in each subject. The procedure was repeated from a more flexed initial position of the elbow (100 degrees). EMG activity from two elbow flexors and two elbow extensors, as well as torque, velocity and joint position were recorded. Healthy control subjects and patients with mild clinical symptoms had coactivation zones or small silent zones around the final positions established after unloading. In these subjects, final positions of the limb were stable. Voluntary movement, i.e., transition of the limb from one initial position to another, was associated with a change in the location of the zone in articular space. The presence of large silent zones in patients with moderate or severe symptoms was correlated with postural instability and oscillations about the final position of the arm after unloading. The comparison of results from healthy and hemiparetic subjects implies that the central specification of the size and the location of a coactivation zone may be fundamental for the control of posture and movement. PMID- 9200499 TI - Brain alpha-adrenoceptors in depressed suicides. AB - alpha1-Adrenoceptors and alpha2-adrenoceptors were measured by radioligand binding to homogenates of brain samples obtained at post-mortem from suicides with a retrospective diagnosis of depression, and age and gender-matched controls. Suicides were subdivided into those who had been free of antidepressant drugs for at least three months, and those in whom prescription of antidepressant drugs was clearly documented. The number of alpha1-adrenoceptors (or alpha1A + alpha1D-adrenoceptors) did not differ significantly between antidepressant-free or antidepressant-treated suicides and controls. In antidepressant-free suicides, the number of alpha2-adrenoceptors was significantly higher in temporal cortex (Ba 21/22). alpha2A-Adrenoceptors did not differ significantly from controls in this brain region, suggesting the involvement of other alpha2-adrenoceptor subtypes. In antidepressant-treated suicides, significantly lower numbers of alpha2-adrenoceptors were found in occipital cortex and hippocampus (and for alpha2A-adrenoceptors in caudate and amygdala) compared to controls. PMID- 9200500 TI - Glutamine transport in cerebellar granule cells in culture. AB - In the present study, uptake of glutamine by rat cerebellar granule cells, a predominantly glutamatergic nerve cell population, has been investigated. Glutamine is taken up by granule cells via at least three transport systems, A, ASC and L. The L-type low affinity system (K(m) = 2.6 mM) is the major transport system in the absence of Na+. The systems A and ASC represent the Na(+)-dependent transport routes, both with almost identical high affinity for glutamine (K(m) = 0.26 mM). Similar transport systems for glutamine are also found in cerebral cortical neurons, a predominantly GABAergic nerve cell population, and cerebral cortical astrocytes. The glutamine transport properties in granule cells, however, show a series of differences from that of cortical neurons and astrocytes: (1) uptake of glutamine by granule cells is primarily mediated by system A (54%), while contributions by system A in cortical neurons and astrocytes are less than 30%; (2) granule cells exhibit strikingly higher transport efficiency for glutamine (V(max)/K(m) = 20 min(-1) for system A as compared to the V(max)/K(m) ratio of 5 min(-1) in cortical neurons and astrocytes), and (3) the initial uptake rates and the steady-state accumulation levels of glutamine are two- to threefold higher in granule cells than that of cortical neurons and astrocytes. These results taken together suggest that in accordance with the important need to replenish the neurotransmitter pool of glutamate, glutamatergic neurons exhibit highly efficient transport systems to accumulate glutamine, one of the major precursors of glutamate. PMID- 9200501 TI - Learning and novelty induced increase of central benzodiazepine receptors from chick forebrain, in a food discrimination task. AB - Young chicks were trained to discriminate food grains from inedible pebbles. On Day 1 and Day 2 of the task, latency to peck, and number of pecks were scored and the forebrain [3H]flunitrazepam receptor binding was also determined at 0 and 30 min after an 8-min training session. Compared with quiet controls, the receptor density increased 46%, 30 min after the training session on Day 1. Compared with chicks that had learned the discrimination and were merely repeating already learned behavior on Day 2, the receptor increased more than 46%. Since chicks that had learned the discrimination had a higher behavioral activity, we interpret that the learning of a new task is itself responsible in addition to stress for the receptor density increase. Stressful factors accompanying the learning task as handling and novelty increased 17% the receptor density, 30 min after a training session without food, compared with quiet controls. However, receptor density did not increase in chicks repeating the same housing conditions, suggesting that chicks were habituated to handling and novelty on Day 2. Differences in receptor density were not observed between quiet controls and experimental groups, at 0 min after the training session, indicating that changes were time dependent. In all cases the affinity remained unchanged. Our results suggest that, the GABA(A) receptor (i) is involved in early stages of memory formation and in stress adaptive responses, and (ii) is modulated by new non repetitive environmental conditions. PMID- 9200502 TI - Inorganic phosphate enhances phosphonucleotide concentrations in cultured fetal rat cortical neurons. AB - Our laboratory has recently characterized saturable Na(+)-dependent P(i) import into cultured fetal rat cortical neurons and shown that a substantial fraction of the P(i) so accumulated is incorporated into ATP. We now report that the ATP, NADPH and intracellular free P(i) ([P(i)]i) concentrations of cultured fetal rat cortical neurons are dependent on the extracellular P(i) concentration ([P(i)]e). [ATP], [NADPH] and [P(i)]i display a hyperbolic dependence upon [P(i)]e, being significantly increased after incubation with [P(i)]e of > or = 10 microM, and maximal at > or = 500 microM. Increases in both [ATP] and [NADPH] are abolished in the absence of glucose. In the absence of extracellular P(i), both [ATP] and [P(i)]i decline over time. Our data suggest that in cultured fetal rat cortical neurons [P(i)]e has a direct effect on glucose utilization, stimulating both ATP and NADPH synthesis via glycolysis and the pentose phosphate pathway. PMID- 9200504 TI - Changes in Fos-like immunoreactivity evoked by maturation of the sneeze reflex triggered by nasal air puff stimulation in kittens. AB - The sneeze reflex is a valuable tool for exploring the maturation of the respiratory control in the newborn as it alters both inspiratory and expiratory activities. Air puff stimulation of the superior nasal meatus innervated by ethmoidal afferents consistently evokes sneeze in adult cats. Such stimulation evokes only a reinforcement of expiratory activities in newborn kittens. This study demonstrates that the pattern of Fos-like immunoreactivity evoked by nasal stimulation changes during functional maturation of sneeze. Nasal stimulation evoked immunoreactivity (i) in the trigeminal sensory complex, at the levels where nasal afferents project, (ii) in the reticular formation, (iii) in the solitary complex and (iv) in the parabrachial area of mature kittens. The evoked immunoreactivity was the same in newborn kittens as in mature kittens in the projection areas of the nasal primary afferents. Fos response was less than half that in mature kittens in the reticular formation and absent in the solitary complex or the parabrachial area. Sneeze can be elicited from the time when evoked immunoreactivity in the solitary complex and the parabrachial area is above control levels. These data provide evidence that the maturation of sneeze is dependent on the development of central relays allowing peripheral inputs to be integrated by neurons engaged in respiratory control. PMID- 9200503 TI - Acute rise in the concentration of free cytoplasmic calcium leads to dephosphorylation of the microtubule-associated protein tau. AB - The objective of this study was to asses the response of the microtubule associated protein tau to acute rise in the concentration of free cytoplasmic calcium ([Ca2+]i) in rat cortical neurons and mouse cerebellar granule cells in culture. One-hour exposure to glutamate (100 microM), N-methyl-D-aspartate (100 microM), KCl (50 mM), and ionomycin (5 microM) led to tau protein dephosphorylation as indicated by an appearance of additional faster moving bands on Western immunoblots with a phosphorylation-independent antibody and an increase in the tau-1 immunoreactivity associated with the appearance of an additional faster moving band. Lowering the extracellular concentration of Ca2+ to less than 1 microM fully prevented the drug-induced tau protein dephosphorylation indicating a dependence on Ca2+ influx from the extracellular environment. Administration of okadaic acid (inhibitor of phosphatase 1/2A) simultaneously with the above mentioned drugs decreased the drug-mediated dephosphorylation. Pre-incubation with okadaic acid fully prevented the dephosphorylation. Treatment with cypermethrin (inhibitor of phosphatase 2B) was without effect when administered either alone, simultaneously with the drugs, or pre-incubated. These findings indicate that, independently of the influx pathway, [Ca2+]i elevation leads to dephosphorylation of the microtubule-associated protein tau and implicate phosphatase 1 and/or 2A in the process. PMID- 9200505 TI - Hypoxia-induced ABR change and heat shock protein expression in the pontine auditory pathway of young rabbits. AB - The auditory brainstem response (ABR) was compared with the immunohistochemical expression of heat shock protein (HSP-72) and microtubule-associated protein 2 (MAP-2) of the brainstem auditory pathway in young rabbits subjected to hypoxic stress. Severe hypoxia for 2 h produced significant prolongation and decreased amplitude of the later component of ABR. HSP-72 expression was distinctly increased in the cochlear nucleus, but there was less induction in the inferior colliculus under severe hypoxia. MAP-2 immunostaining of neuropiles in the inferior collicular nucleus was decreased slightly after severe-long hypoxia, but cytoplasmic staining did not change. The present ABR change, which was produced by brainstem hypoxia-ischemia and acidosis, may be due to the neural cytoarchitectural derangement and less induction of stress proteins in the upper brainstem. PMID- 9200506 TI - The effects of traumatic brain injury on inhibition in the hippocampus and dentate gyrus. AB - Changes in inhibitory neuronal functioning may contribute to morbidity following traumatic brain injury (TBI). Evoked responses to orthodromic paired-pulse stimulation were examined in the hippocampus and dentate gyrus at 2 and 15 days following lateral fluid percussion TBI in adult rats. The relative strength of inhibition was estimated by measuring evoked paired pulses in three afferent systems: the CA3 commissural input to the CA1 region of the hippocampus; the entorhinal cortical input to the ipsilateral CA1 area (temporoammonic system); and the entorhinal input to the ipsilateral dentate gyrus (perforant path). In addition to quantitative electrophysiological estimates of inhibitory efficacy, levels of gamma-aminobutyric acid (GABA) were qualitatively examined with immunohistochemical techniques. Effects of TBI on paired-pulse responses were pathway-specific, and dependent on time postinjury. At 2 days following TBI, inhibition of population spikes was significantly reduced in the CA3 commissural input to CA1, which contrasted with injury-induced increases in inhibition in the dentate gyrus seen at both 2 and 15 days postinjury. Low-level stimulation, subthreshold for population spikes, also revealed changes in paired-pulse facilitation of field extracellular postsynaptic potentials (fEPSPs), which depended on fiber pathway and time postinjury. Significant injury-induced electrophysiological changes were almost entirely confined to the hemisphere ipsilateral to injury. Intensity of GABA immunobinding exhibited a regional association with electrophysiological indices of inhibition, with the most pronounced increases in GABA levels and inhibition found in the dentate gyrus. TBI-induced effects showed a regional pattern within the hippocampus which corresponds closely to inhibitory changes reported to follow ischemia and kindling. This degree of similarity in outcome following dissimilar injuries may indicate common mechanisms in the nervous system response to injury. PMID- 9200507 TI - Antinociception induced by opioid or 5-HT agonists microinjected into the anterior pretectal nucleus of the rat. AB - The changes in the latency for tail withdrawal in response to noxious heating of the skin induced by microinjection of opioid or serotonergic agonists into the anterior pretectal nucleus (APtN) was studied in rats. The mu-opioid agonist DAMGO (78 and 156 picomol), but not the delta-opioid agonist DADLE (70 and 140 pmol), the kappa-opioid agonist bremazocine (0.24 and 0.48 nanomol) or the sigma opioid agonist N-allylnormetazocine (0.54 nanomol), produced a dose-dependent antinociceptive effect. The 5-HT1 agonist 5-carboxamidotryptamine (19 and 38 nanomol) and the 5-HT1B agonist, CGS 12066B (1.12 and 2.24 nanomol), but not the non-selective 5-HT agonist m-CPP (41 to 164 nanomol), 5-HT2 agonist alpha methylserotonin (36 and 72 nanomol) and 5-HT3 agonist 2-methylserotonin (36 and 72 nanomol), produced a dose-dependent antinociceptive effect. These results indicate that the antinociceptive effects of opioid or serotonergic agonists microinjected into the APtN depend on drug interaction with local mu or 5-HT1B receptors, respectively. PMID- 9200508 TI - The influence of preischemic hyperglycemia on acute changes in the apparent diffusion coefficient of brain water following global ischemia in rats. AB - We report the effect of increased plasma glucose levels on changes in the apparent diffusion coefficient of brain water (ADCw) during the first few minutes of global ischemia in rats. Brain ADCw values were acquired every 15 s using a diffusion-weighted line-scan MR pulse sequence. Preischemic hyperglycemia was achieved by infusion of 50% dextrose (i.v.) prior to KCl-induced cardiac arrest global ischemia. Analysis based on single voxels (3.4 microl) in brain demonstrated significant differences in the time course of ADCw decline between normoglycemic (n = 8) and hyperglycemic (n = 6) groups. Mean data from the hyperglycemic group indicated a biphasic decline of ADCw that was characterized by an initial rapid drop followed by a plateau of approximately 1 min before gradually declining and leveling off to its minimum value. In the normoglycemic group, ADCw declined to the same value as in the hyperglycemic group, but without a notable plateau. In the cerebral cortex, the times to maximal and half maximal ADCw drop following global ischemia in the hyperglycemic group were 3.96 and 2.26 min respectively. Corresponding time intervals for the normoglycemic group were 1.86 and 1.14 min, respectively. The time course for changes in ADCw demonstrated here is significantly different than that for anoxic depolarization reported under similar experimental conditions and suggests that events other than the complete loss of membrane ionic homeostasis and subsequent cell swelling may be involved in the initial decline of ADCw in global cerebral ischemia. PMID- 9200509 TI - Brain-gut induction of heat shock protein (HSP) 70 mRNA by psychophysiological stress in rats. AB - Restraint water-immersion stress-induced expression of heat shock protein (HSP)70 mRNA in the cerebral cortex and stomach of rats was evaluated by Northern blotting. Cerebral and gastric HSP70 mRNA significantly increased in the 6 h stressed rats and the amount of mRNA measured as optical densities was highest in the 12 h-stressed rats. These data confirmed our previous observations and suggest that families of HSPs play a salient cytoprotective role in stress vulnerable organs. PMID- 9200510 TI - Mechanical stimulation of neurites generates an inward current in putative aortic baroreceptor neurons in vitro. AB - We investigated the responses of putative aortic baroreceptor neurons to mechanical stimulation of their processes. Putative aortic baroreceptor neurons were identified by applying the carbocyanine dye DiI to the adventitia of the aortic arch of anesthetized rats. After at least 1 week, the nodose ganglia were removed and the neurons were cultured. Within 2-3 days, neurite outgrowth was evident on many neurons. The soma was voltage-clamped using whole cell patch clamp techniques while the neurites were deformed with pneumatic ejection of bath solution at 5-15 psi using a glass pipette (7-15 microm) positioned at least 50 microm from the neurite. Mechanical stimulation induced an inward current in 15 out of 17 putative aortic baroreceptor neurons. The magnitude of the current was related to the intensity of stimulation. The current was blocked by 20 microM gadolinium (n = 11), a reported blocker of mechanically sensitive ion channels, or by incubating the cells overnight in 10 microM phalloidin, which binds to actin filaments (n = 5). We conclude that mechanical deformation of neurites of putative baroreceptor neurons activates a mechanosensitive inward current in the soma and that the cytoskeletal actin filaments are involved in the generation of this current. PMID- 9200511 TI - Cardiovascular effects of nitric oxide in the rostral ventrolateral medulla of rats. AB - To investigate the cardiovascular role of nitric oxide (NO) in the rostral ventrolateral medulla (RVLM), NOC 18, an NO donor, was microinjected into the RVLM of rats. NOC 18 significantly decreased mean arterial pressure (MAP). Pre treatment with an NO trapper, carboxy-PTIO, abolished the NOC 18-induced decrease in MAP. Microinjection of L-NAME, an NO synthase inhibitor, increased MAP. L Arginine reduced MAP and inhibited the pressor response induced by L-NAME. Results suggest that NO acts on the RVLM neurons and plays an important role in cardiovascular regulation. PMID- 9200513 TI - Gas chromatography in anaesthesia. I. A brief review of analytical methods and gas chromatographic detector and column systems. AB - Practical applications and relevant studies involving the anaesthetic gases, have been extensively described in the literature. Many eminent analytical methods have already been developed for medical practice where routine analysis of anaesthetics is frequently needed, particularly during anaesthesia, and in related and respiratory research programmes. The determination of halothane, isoflurane, enflurane and nitrous oxide concentrations from vaporizers, in exhaled and inhaled gas mixtures, in body fluids and tissues is necessary to control anaesthetic concentrations, and thus, the relevant and adverse effects successfully. Therefore, a literature review, with particular emphasis on gas chromatography would provide important information for investigators in the search for a suitable analytical method for the analysis of multi-component mixtures of anaesthetic gases. PMID- 9200512 TI - Immunoreactivity of presenilin-1 in human, rat and mouse brain. AB - Monoclonal antibodies (mAbs) D3G6 and C8A5, specific for amino acid residues 160 168 of S182 protein, immunolabeled neurons, ependymal and choroid plexus cells, and myocytes in brain sections from normal subjects and people with Alzheimer disease or Down syndrome and in rats and mice. Oligodendroglia, microglia, and the majority of astrocytes were negative. S182 protein or a fragment of the protein detected with these mAbs is not a constituent of amyloid-beta deposits or tangles. PMID- 9200514 TI - Determination of 3-oxo-delta4- and 3-oxo-delta4,6-bile acids and related compounds in biological fluids of infants with cholestasis by gas chromatography mass spectrometry. AB - A method has been developed for the determination of 3-oxo-delta4- and 3-oxo delta4,6-bile acids and related bile acids in biological fluids of infants by gas chromatography-mass spectrometry (GC-MS) of the methyl ester-dimethylethylsilyl ether-methoxime derivatives. The 7alpha-hydroxylated 3-oxo-delta4-bile acids were partially dehydrated to give the 3-oxo-delta4,6-bile acids by trimethylsilyl or dimethylethylsilyl derivatization and other pretreatments under acidic or alkaline conditions for GC-MS analysis. To prevent dehydration, the 3-oxo-delta4 bile acids were derivatized to the oximes by treatment with O-methylhydroxylamine prior to pretreatments such as solid-phase extraction, enzymatic solvolysis and hydrolysis of the conjugates, and silylation with dimethylethylsilylimidazole. Calibration curves for the bile acids were linear over a range of 5-250 ng and the detection limit was 100 pg for each 3-oxo-delta4-bile acid. Recoveries of the bile acids and their glycine and taurine conjugates from bile acid-free urine and serum ranged from 94.2 to 105.9% of their added amounts. The bile acids in urine and serum of four patients with severe cholestatic liver disease were measured by the analytical method, and the 3-oxo-delta4-bile acids were determined to be the major bile acids (59-68%) in the urines associated with 3-oxo-delta4-steroid 5beta-reductase deficiency or dysfunction. PMID- 9200515 TI - High-performance liquid chromatographic determination of biogenic amines in fish implicated in food poisoning. AB - A rapid, sensitive and reproducible high-performance liquid chromatographic procedure for the determination of nine biogenic amines in fish by improved benzoylation with benzoyl chloride was developed. The benzoylation of amines with benzoyl chloride at 30 degrees C for 40 min was the optimal condition to eliminate the influence of interfering peaks during analysis. The calibration curve for each amine was linear within the range of 0.02-4 microg. The amine recovery from fish meat was better by extraction with 6% trichloroacetic acid than with 1 M HClO4. The application of this method to detect amines in a fried marlin fillet implicated in a food poisoning incident indicated that a high level (84.1 mg/100 g) of histamine was present in the sample. PMID- 9200516 TI - High speed liquid chromatography of phenylethanolamines for the kinetic analysis of [11C]-meta-hydroxyephedrine and metabolites in plasma. AB - A method is developed and described for analysis of [11C]-meta-hydroxyephedrine, [11C]MHED, a tracer of cardiac function, and its metabolites in plasma samples. The method combines on-column solid-phase extraction and separation on a single weak cation-exchange column. Phenylethanolamines were used to develop the separation method that concentrates the analytes on-column from physiological saline and then elutes them by changing to an acidic mobile phase. Hydrophobic interactions determine the selectivity, and elution order is the same as for reversed-phase liquid chromatography on a C1 stationary phase. The mechanism of separation is mixed mode, with ion-exchange coupled with a reversed-phase liquid chromatography mechanism. Each sample analysis requires only 10 min and does not require deproteinization or the use of organic solvents. In human samples, a single plasma metabolite of [11C]MHED along with the parent compound were observed using this method. The method was sufficiently rapid so that in 70 min seven samples were assayed, providing a well-defined time course for MHED and its metabolites in blood. The metabolite concentration increased with time to approximately 85% of the plasma activity 50 min after administration. The results with the developed method are comparable to those described for reversed-phase separations, with the advantage that our method does not require deproteinization, reducing sample analysis time by a factor of two. PMID- 9200517 TI - Ion chromatographic study of sodium, potassium and ammonium in human body fluids with bulk acoustic wave detection. AB - In the present paper, determination of Na+, K+ and NH4+ in saliva and serum was carried out using an ion chromatography (IC) method with a bulk acoustic wave (BAW) sensor as detector and 2.0 mmol/l nitric acid as mobile phase. The IC-BAW method is simple, rapid and accurate. Comparison between the BAW detector and the conventional conductivity detector (CDD) was discussed. The IC-BAW showed agreement with the commonly used flame photometric method for Na+ and K+ and enzymatic method for NH4+, as well as IC-CDD for those three cations. PMID- 9200518 TI - Highly sensitive gas chromatographic-mass spectrometric method for morphine determination in plasma that is suitable for pharmacokinetic studies. AB - A sensitive method was devised to determine morphine plasma concentrations by gas chromatography-mass spectrometry (GC-MS) using selected ion monitoring (SIM) with nalorphine as the internal standard. This method was rugged, reliable, selective and sensitive and was used to determine the morphine content of over 2000 samples. Sample preparation involved extraction of basified sample using n-butyl chloride-chloroform (5:1) and evaporation of the extract to dryness. The residue was derivatised with pentafluoropropionic anhydride, evaporated to dryness, reconstituted in 40 microl toluene and injected onto the GC-MS. For a sample size of 1 ml, the limit of quantitation was 0.75 ng/ml (S/N ratio 10:1) and the estimated limit of detection was calculated to be 0.2 ng/ml (S/N ratio 3:1), expressed as morphine base. Precision (n=5) was 4.9% at 0.75 ng/ml, 6.8% at 1.5 ng/ml, 3.0% at 37.5 ng/ml and 2.3% at 150 ng/ml. Standard curves for the range of 0-750 ng/ml morphine in plasma were linear with all r2 values greater than 0.99. No interfering peaks were seen for either morphine or internal standard in the blank samples. The method is suitable for pharmacokinetic studies after subclinical doses of morphine where it has been used to study morphine plasma concentrations for 6 h after a dose of only 2 mg. PMID- 9200519 TI - Gas chromatographic-mass spectrometry and gas chromatographic-Fourier transform infrared spectroscopy assay for the simultaneous identification of fentanyl metabolites. AB - Fentanyl, a synthetic opioid, undergoes important biotransformation to several metabolites. A gas chromatographic-mass spectrometric assay was applied for the simultaneous analysis of fentanyl and its major metabolites in biological samples. The identification of different metabolites was performed by gas chromatography-mass spectrometry (electronic impact and chemical ionisation modes) and gas chromatography-Fourier transform infrared spectroscopy. In the present study, rat and human microsomes incubation mixtures and human urines were analysed. In vitro formation of already known fentanyl metabolites was confirmed. The presence of metabolites not previously detected in human urine is described. PMID- 9200520 TI - Gas chromatographic method using electron-capture detection for the determination of musk xylene in human blood samples. Biological monitoring of the general population. AB - Musk xylene (2,4,6-trinitro-1,3-dimethyl-5-tert.-butylbenzene, MX), a synthetic musk often used in different fragrances and soaps to substitute the natural musk, is a potential contaminant of humans. In this publication, a specific and sensitive detection method for the determination of musk xylene in human blood samples is described. The clean-up of the blood samples includes an extraction step followed by a solid-phase adsorption to separate MX from other plasma components. Separation and detection was carried out by capillary gas chromatography and an electron capture detector (GC-ECD). The results were verified using qualitative capillary gas chromatography and a mass selective detector with electron impact ionisation (GC-EI-MS). epsilon Hexachlorocyclohexane (epsilon-HCH) is used as internal standard. The reliability of the GC-ECD method has been proved. The relative standard deviations of the within-series imprecision were 12.7% for samples with a concentration of 0.5 microg/l and 2.1% for samples with a concentration of 5.0 microg/l, whereas the relative standard deviations for the between-day imprecision were 14.9% (0.5 microg/l samples) and 3.4% (5.0 microg/l samples). The losses during sample treatment were between 10.1% and 17.8%. No interfering peaks were observed. The absolute detection limit was 0.1 microg/l plasma. A total of 72 human blood samples were analysed to determine the MX concentrations within the general population. In 66 of the 72 human blood samples, the MX concentrations ranged from 0.10 to 1.12 microg/l plasma for the described method. In six samples no MX was detected. The median concentration was 0.24+/-0.23 microg MX/l plasma. The 95 percentile was 0.79 microg/l. No correlation could be found between MX concentrations and smoking habit, broca index, age, sex as well as fish consumption habits. Nevertheless, the results demonstrate the exposure of the general population to MX. PMID- 9200521 TI - Membrane adsorbers for selective removal of bacterial endotoxin. AB - Surface-modified flat-sheet microfiltration membranes were functionalised with poly-L-lysine, polymyxin B, poly(ethyleneimine), L-histidine, histamine, alpha amylase and DEAE as well as deoxycholate. Their suitability to remove endotoxin from both buffers and protein solutions was examined using bovine serum albumin, murine IgG1 and lysozyme as model proteins. In protein-free solutions reduction from 6000 EU/ml to <0.1 EU/ml was achieved with all applied ligands; only alpha amylase as well as L-histidine and histamine, when immobilized via the non-ionic spacer bisoxirane, exhibited low clearance factors at neutral pH. The adsorption of endotoxin is mainly ruled by electrostatic interaction forces. Thus in multi component systems, such as endotoxin-contaminated protein solutions, competing interactions take place: acidic proteins compete with endotoxin for binding sites at the membrane adsorbers, basic proteins compete with the ligands for endotoxin and act as endotoxin carriers. With properly chosen conditions the membrane adsorbers presented here show exceptional effectiveness also in the presence of proteins. They are generally superior to functionalised Sepharose chromatographic sorbents and allow fast processing. They may contribute to reduce the risks in the application of parenterals and diagnostics. PMID- 9200522 TI - Sensitive microanalysis of imipramine and desipramine in single rat thyroids by gas chromatography-mass spectrometry. AB - A new sensitive method for the quantitative determination of imipramine and desipramine in single rat thyroids using gas chromatography-mass spectrometry with selected ion monitoring, after enzymatic hydrolysis and liquid-liquid extraction has been developed. The technique was deemed suitable for microanalysis of single rat thyroids and for other solid tissues, using smaller sample sizes than usually required for traditional determination methods. The quantification was linear from 10 to 200 nmol/l (i.e., from 0.25 to 5 microg/g) for imipramine and from 100 nmol/l to 2000 nmol/l (i.e., from 2.4 to 47 microg/g) for desipramine, and the limits of detection (less than 25 ng/g tissue for both compounds) were better than those previously reported. Recoveries, repeatability and reproducibility of this technique were satisfactory. It has been successfully applied in a preliminary study of the concentration-time profiles of imipramine and desipramine in the thyroid of rats treated with either of these drugs. PMID- 9200523 TI - Lamotrigine analysis in plasma by gas chromatography-mass spectrometry after conversion to a tert.-butyldimethylsilyl derivative. AB - Lamotrigine (lamictal) is a new anticonvulsant drug recently approved by the FDA for clinical use. Therapeutic monitoring of lamotrigine is useful for patient management (therapeutic range 1-4 microg/ml). Here we describe a gas chromatography-mass spectrometric identification and quantitation of lamotrigine after extraction from human serum and derivatization. Lamotrigine was extracted from alkaline serum with chloroform and derivatized with N-methyl-N-(tert. butyldimethysilyl) trifluoroacetamide containing 2% tert. butyldimethylchlorosilane. Oxazepam-d5 was used as an internal standard. The tert.-butyldimethylsilyl derivative of lamotrigine showed distinct molecular ions at m/z 483 and 485 as well as other peaks at m/z 426, 370 and 334 for unambiguous identification. The base peak was observed at m/z 199. Similarly, the tert. butyldimethysilyl derivative of oxazepam-d5 showed molecular ions at m/z 519 and 521 along with other characteristic peaks at m/z 462, 376 and 318. For the analysis of lamotrigine, the mass spectrometer was operated in the selective ion monitoring mode. The within-run and between-run precisions were 4.3% (mean=3.01, S.D.=0.13 microg/ml) and 5.1% (mean=2.93, S.D.=0.15 microg/ml), respectively at a serum lamotrigine concentration of 3.0 microg/ml. The within-run and between-run precisions were 8.2% (mean=0.49, S.D.=0.04 microg/ml) and 10.6% (mean=0.47, S.D.=0.05 microg/ml), respectively at a serum lamotrigine concentration of 0.5 microg/ml. The assay was linear for serum lamotrigine concentrations of 0.5-20 microg/ml. The detection limit was 0.25 microg/ml. The assay was free from interferences from common tricyclic antidepressants, benzodiazepines, other common anticonvulsants, salicylate and acetaminophen. PMID- 9200524 TI - Quantitative determination of BMS186716, a thiol compound, in dog plasma by high performance liquid chromatography-positive ion electrospray mass spectrometry after formation of the methyl acrylate adduct. AB - As it is extremely unstable in blood, the thiol compound BMS186716 was stabilized by the addition of methyl acrylate (MA) to blood samples. The blood samples were then kept in ice for 10-15 min for completion of the Michael addition reaction to occur between the thiol group of BMS186716 and MA, after which the plasma was separated by centrifugation under refrigeration. For sample analysis, the standard and quality control samples were prepared by spiking blank plasma with the BMS186716-MA adduct. After addition of the internal standard, BMS 188035-MA, each sample was acidified with HCI and then extracted with methyl tert.-butyl ether. Each reconstituted extract was injected into a high-performance liquid chromatography-positive ion electrospray ionization mass spectrometric system. The electrospray condition was chosen to enhance the [M+NH4]+ signal at the expense of the [M+H]+ signal. Monitoring the [M+NH4]+ signal, a lower limit of quantitation of 2.5 ng/ml was achieved, with 0.5 ml plasma. We have thus shown that a sulfhydryl compound (BMS186716) in blood can successfully be stabilized by reacting it with MA and that the adduct produced is adequately stable in blood and plasma to allow the development of a rugged quantitative bioanalytical method. PMID- 9200525 TI - Low level determination of a novel 4-azasteroid and its carboxylic acid metabolite in human plasma and semen using high-performance liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry. AB - Compound I (4.7beta-dimethyl-4-azacholestan-3-one, MK-0386) is a potent 5alpha reductase type 1 (5alphaR1) inhibitor. Sensitive (0.2 ng/ml), specific and separate assays have been developed and validated for the analysis of I and its carboxylic acid metabolite (II) in human semen and plasma based on high performance liquid chromatography (HPLC) with tandem mass spectrometric (MS-MS) detection. After liquid-liquid extraction of the analytes from biological matrix, the extracts were chromatographed on a short (50 mm) analytical column during analysis of I, and on a longer (150 mm) column with a weaker mobile phase during the analysis of II. This additional chromatographic separation was required to separate II from a secondary metabolite present in post-dose plasma samples interfering with the quantification of II. The MS-MS detection was performed on a Sciex API III Plus tandem mass spectrometer using the heated nebulizer probe. Monitoring the parent-->product ion combinations of m/z 416-->114 and 404-->114, in the multiple reaction monitoring (MRM) mode, after chromatographic separation, allowed quantification of both analytes. The standard curve in plasma was linear in the concentration range of 0.2 to 200 ng/ml for both I and II with correlation coefficients greater than 0.99 and coefficients of variation of less than 15% for replicate (n=5) analysis at all concentrations within the standard curve range. For the semen assay the linear range for determination of I was from 0.2 to 50 ng/ml. These assays were applied to support a number of clinical studies with I and their validity and long-term performance was confirmed during analyses of clinical samples from these studies. The need for careful assessment of the specificity of MS-MS assays in post-dose biological fluid samples in the presence of metabolites was emphasized. PMID- 9200526 TI - Determination of a novel growth hormone secretagogue (MK-677) in human plasma at picogram levels by liquid chromatography with atmospheric pressure chemical ionization tandem mass spectrometry. AB - A sensitive and specific method for the determination of N-[1(R)?[1,2-dihydro-1 methylsulfonylspiro(3H-indole-3,4'-piper idin)-1'-yl]carbonyl?-2 (phenylmethoxy)ethyl]-2-amino-2-meth ylpropanamide (MK-677, I), a growth hormone secretagogue, has been developed. The method is based on high-performance liquid chromatography (HPLC) with tandem mass spectrometric (MS-MS) detection. The analyte and internal standard (II) were isolated from the basified plasma using a liquid-liquid extraction with methyl-tert.-butyl ether (MTBE). The organic extract was evaporated to dryness, the residue was reconstituted in mobile phase and injected into the HPLC system. The MS-MS detection was performed on a PE Sciex API III Plus tandem mass spectrometer using a heated nebulizer interface. Multiple reaction monitoring of parent-->product ion combinations at m/z 529- >267 and 527-->267 was used to quantify I and II, respectively. The assay was validated in human plasma in the concentration range of 0.1 to 100 ng/ml, and the limit of quantification (LOQ) was 0.1 ng/ml. The precision of the assay, as expressed as coefficients of variation (C.V.,%) was less than 7% at all concentrations within the standard curve range, with adequate assay specificity and accuracy. The HPLC-MS-MS method provided sufficient sensitivity to completely map the pharmacokinetic time-course following a single 5-mg oral dose of I. PMID- 9200527 TI - Amperometric detection of organic thiols at a tungsten wire electrode following their separation by liquid chromatography. AB - The amperometric detection of thiols following their separation by reversed-phase chromatography and reaction into a post-column mercury carrier stream is shown to be a sensitive method when using a tungsten wire sensor as the working electrode in a three-electrode flow cell. Five organic thiols (cysteine, homocysteine, reduced glutathione, D,L-penicillamine and 2-mercaptopropionic acid) and thiourea could be separated within approx. 18 min. The analytical performance is comparable, and stability superior, to chemically modified electrodes previously reported. PMID- 9200528 TI - Quantitative determination of paroxetine in plasma by high-performance liquid chromatography and ultraviolet detection. AB - An accurate, reliable procedure was developed for kinetic and therapeutic monitoring of paroxetine in human plasma. Steady-state plasma levels of paroxetine were measured for 18 geriatric patients (mean age 75) in a double blinded study. Paroxetine doses ranged from 10 to 40 mg/day. The assay was suitable for patients on concurrent medications, and a small sample volume (1 ml) of patient plasma was used with sufficient sensitivity and specificity. After extraction and separation on a Beckman, Ultrasphere 5-microm C18 column (150x2 mm I.D.), the recovery (mean+/-S.D.) for paroxetine was determined to be 86.5+/ 5.2%. The limit of quantitation for paroxetine in this assay was 5 ng/ml. Inter assay reproducibility (C.V.) for the patient samples and quality controls ranged from 3.7 to 7.6%. PMID- 9200529 TI - Simultaneous determination of imipramine, desipramine and their 2- and 10 hydroxylated metabolites in human plasma and urine by high-performance liquid chromatography. AB - A simultaneous assay for imipramine, desipramine and their 2- and 10-hydroxy metabolites using high-performance liquid chromatography (HPLC) is described. The drugs and internal standard, pericyazine, were extracted from plasma or urine at pH 9.6 with diethyl ether and back-extracted into 0.1 M orthophosphoric acid. The recovery of the compounds ranged from 78.6% for imipramine to 94.3% for 2 hydroxydesipramine. The extracts were analysed by reversed-phase HPLC with electrochemical detection using a mobile phase of 30% acetonitrile in 0.1 M K2HPO4 at pH 6.0 delivered at 2 ml/min. All compounds were resolved in a run time of 15 min with lower limits of quantification of 1.5 ng/ml for hydroxy metabolites and 3 ng/ml for imipramine and desipramine. The intra- and inter-day coefficients of variation at 50 ng/ml were 5.2% and 6.8%, respectively (n=8). PMID- 9200530 TI - Determination of two CI-1007 sulfate metabolites in monkey plasma and urine. AB - Two HPLC assays were developed and validated for simultaneous quantitation of two sulfate metabolites, PD 163637 (VI) and PD 163639 (VIII), of an investigational antipsychotic drug CI-1007 (I) in monkey plasma and urine. VI and VIII were identified as major metabolites in monkey plasma, and both were excreted in urine. Monkey plasma samples were directly injected after deproteinization, and urine samples were analyzed after a clean-up procedure using methyl-tert.-butyl ether. Liquid chromatographic separation was achieved on a Zorbax RX C8 analytical column using gradient elution. Column effluent was monitored using fluorescence detection with excitation and emission wavelengths of 254 and 330 nm, respectively. Minimum quantitation limit was 50 ng/ml in plasma and 100 ng/ml in urine. Linearity was demonstrated up to 3000 ng/ml in plasma and urine. Recoveries of the analytes from plasma and urine were greater than 85%. The assay has been applied to the determination of VI and VIII in plasma and urine samples from monkeys receiving oral administration of I. PMID- 9200531 TI - High-performance liquid chromatographic assay using electrochemical detection for the combined measurement of amifostine, WR 1065 and the disulfides in plasma. AB - A high-performance liquid chromatographic (HPLC) method was developed for the combined analysis of the chemoprotective agent, amifostine, its active metabolite, WR 1065, and the (symmetrical and mixed) disulfides of WR 1065 in plasma. These three compounds were quantified by measuring WR 1065 after three different sample pretreatment procedures. During these procedures, amifostine and the disulfides were quantitatively converted into WR 1065, by incubating the sample either at a low pH or in the presence of dithiothreitol, respectively. The resulting amounts of WR 1065 were determined by HPLC with electrochemical detection (Au electrode, + 1.00 V). The lower limit of quantitation of WR 1065 was 0.15 microM. The within-day and between-day precision were < or =4.4 and < or =8.2% for WR 1065, < or =4.9 and < or =13.1% for amifostine and < or =8.5 and < or =5.5% for the disulfides, respectively. The within-day and between-day accuracy ranged from 97.2 to 109.8% and from 97.6 to 101.5% for WR 1065, from 88.3 to 110.7% and from 99.4 to 101.5% for amifostine and from 99.2 to 110.2% and from 103.3 to 104.9% for the disulfides, respectively. This method is superior to other described methods due to its simple and relatively rapid analysis of all three compounds in one system. Furthermore, it is at least as sensitive as earlier reported methods for one of the compounds and the application of the gold electrode requires only minor maintenance. Therefore, this method is very suitable for pharmacokinetic studies of amifostine and its metabolites. As an example, the plasma concentrations of amifostine, WR 1065 and the disulfides are shown in a patient after receiving an i.v. dose of 740 mg/m2 amifostine. PMID- 9200532 TI - High-performance liquid chromatographic determination of cisplatin as platinum(II) in a pharmaceutical preparation and blood samples of cancer patients. AB - An high-performance liquid chromatographic (HPLC) method has been developed for the determination of cisplatin, based on precolumn derivatization of platinum(II) with bis(salicylaldehyde)tetramethylethylenediimine, extraction in chloroform and elution from a 3 microm Hypersil ODS column with methanol-acetonitrile-water as mobile phase and detection at 254 nm. Copper(II), iron(II), nickel(II), palladium(II), dioxouranium(IV) separated completely and did not affect the determination of platinum(II). The method was applied for the determination of cisplatin as platinum(II) in a pharmaceutical preparation and in blood samples of cancer patients after infusion of cisplatin. PMID- 9200533 TI - Bioanalysis of captopril: two sensitive high-performance liquid chromatographic methods with pre- or postcolumn fluorescent labeling. AB - This study describes the development and comparison of two HPLC methods for the analysis of the antihypertensive drug captopril. The first method is based on a precolumn derivatization of captopril with the fluorescent label monobromobimane (MBB). The second method is based on a postcolumn reaction with the fluorescent reagent o-phthaldialdehyde (OPA). Since the disulfide metabolites of captopril can be reconverted to the active drug in vivo, the bioanalysis of captopril should involve both the determination of its free thiol form (free captopril) and the total amount of free thiol and reducible disulfides (total captopril). For total captopril analysis, disulfides were reduced with tributylphosphine (TBP) prior to protein precipitation. Since the reducing agent interfered with the MBB derivatization reaction, this method was not suitable for total captopril analysis. Both methods were validated for the bioanalysis of free captopril in human plasma. After removal of plasma proteins, samples were analyzed without an additional extraction procedure. The limit of quantitation in plasma was 12.5 ng/ml for the MBB method (limit of detection 30 pg) and 25 ng/ml for the OPA method (limit of detection 50 pg). The OPA method was also validated for total captopril analysis in human plasma and urine. The limit of quantitation was 25 ng/ml in plasma and 250 ng/ml in urine (limit of detection 50 pg). We conclude that for the analysis of free captopril the precolumn MBB method is superior to the OPA method since only the derivatization reaction has to be carried out immediately. The postcolumn OPA method is especially suitable for the analysis of total captopril since reducing reagents and high concentrations of endogenous thiols do not interfere with the derivatization reaction. PMID- 9200534 TI - Chromatographic identification of phenolic compounds in human urine following oral administration of the herbal medicines Daisaiko-to and Shosaiko-to. AB - Chemical identification of the compounds in human urine following administration of the traditional Chinese medicines, Daisaiko-to and Shosaiko-to (Dachaihu-tang and Xiaochaihu-tang in Chinese, respectively), was achieved by using a linear relationship between the logarithm of the capacity factor, log k', and that of the volume fraction of CH3CN, log X(s)(vol), in the aqueous mobile phase: -log k'=A+B log X(s)(vol). Comparison of the slope, B, and the intercept, A, between the urinary compound and its suspected authentic specimen gave satisfactory results in the chemical identification. We applied this method to the initial stage of pharmacokinetic studies on the herbal medicines and identified seven flavonoids and two anthraquinone derivatives in the urine specimens obtained after herbal administration. PMID- 9200536 TI - Stereoselective determination of R-(-)- and S-(+)-prilocaine in human serum by capillary electrophoresis using a derivatized cyclodextrin and ultraviolet detection. AB - A capillary electrophoresis (CE) method for the quantification of R-(-)- and S (+)-prilocaine in human serum was developed and validated. Stereoselective resolution was accomplished using 15 mM heptakis(2,6-di-methyl)-beta-cyclodextrin and 0.03 mM hexadecyltrimethylammonium bromide (HTAB) contained in 100 mM phosphate buffer, pH 2.5. Solid-phase extraction was used as a sample preparation technique to remove endogenous interferences. A 72-cm uncoated fused-silica capillary at a voltage of 25 kV and 30 degrees C was used for the analysis. The detection limits for R-(-)- and S-(+)-prilocaine were 38 ng/ml using 1 ml of human serum and the limits of quantitation were 45 ng/ml. The calibration curve was linear over the range of 45-750 ng/ml with procainamide as the internal standard. Precision and accuracy of the method were 2.86-8.50% and 3.29-7.40%, respectively, for R-(-)-prilocaine, and 3.94-9.17% and 2.0-6.73%, respectively, for S-(+)-prilocaine. The CE method was compared to an existing chiral HPLC method in terms of sensitivity and selectivity for the routine analysis of the drug. PMID- 9200537 TI - Simultaneous determination of phospholipid hydroperoxides and cholesteryl ester hydroperoxides in human plasma by high-performance liquid chromatography with chemiluminescence detection. AB - A method was developed for the simultaneous determination of phosphatidylcholine hydroperoxides (PCOOH) and cholesteryl ester hydroperoxides (CEOOH). Lipid hydroperoxides (LOOH) were quantitatively extracted from human plasma with a mixture of n-hexane and ethyl acetate, and separated by column-switching high performance liquid chromatography using one aminopropyl column and two octyl columns followed by chemiluminescence detection. LOOHs could be completely separated from each other and detected at picomole levels. The results of method validation tests were satisfactory. This method was then applied to determine LOOH in normal human plasma; the levels of PCOOH and CEOOH found were 36.0+/-4.0 nM (mean+/-S.D., n=6) and 12.3+/-3.1 nM (mean+/-S.D., n=6), respectively. PMID- 9200535 TI - High-performance thin-layer chromatographic method for the detection and determination of lansoprazole in human plasma and its use in pharmacokinetic studies. AB - A rapid and sensitive high-performance thin-layer chromatography (HPTLC) method has been developed for the measurement of lansoprazole in human plasma and its use for pharmacokinetic study has been evaluated. Detection and quantitation were performed without using an internal standard. A single stage extraction procedure was followed for extracting lansoprazole from plasma and a known amount of the extract was spotted on precoated silica gel 60 F254 plates using a Camag Linomat IV autosampler. Lansoprazole was quantified using a Camag TLC Scanner 3. The recovery study of authentic analytes added to plasma at 0.05 to 0.25 microg/ml was 95.37+/-2.15% and the lowest amount of lansoprazole that could be detected was 20 ng/ml plasma. The method provides a direct estimate of the amount of lansoprazole present in plasma. The method was used for the determination of plasma levels as well as pharmacokinetic parameters of lansoprazole after oral administration of two marketed preparations to healthy volunteers. PMID- 9200538 TI - High-performance liquid chromatographic determination of myocardial interstitial dihydroxyphenylglycol. AB - This study describes a high-performance liquid chromatographic method with electrochemical detection (HPLC-ED) for monitoring dihydroxyphenylglycol (DHPG) in the myocardial interstitial space. Using a cardiac dialysis technique, 10 microl dialysates were sampled from the myocardial interstitial space (1-min fractions) and were injected directly into the HPLC-ED system. The in vitro quantification limit for DHPG was 250 fg in a 10-microl injection. The basal DHPG concentration of dialysate was 181+/-46 pg/ml. This system offers a new possibility for monitoring myocardial interstitial DHPG levels. PMID- 9200539 TI - Determination of tramadol in human plasma by capillary gas chromatography-mass spectrometry using solid-phase extraction. AB - An analytical method using solid-phase extraction, capillary gas chromatography and mass selective detection in the electron-impact ionization (EI) mode was developed for the determination of tramadol (2-[(dimethylamino)methyl]-1-(3 methoxyphenyl)cyclohexanol) in human plasma. The advantages of this method are the high sensitivity and selectivity and the linearity over the concentration range 2-500 ng/ml. Quantification was made using nefopam as an internal standard, and the detection limit was found to be 1 ng/ml. The standard deviations of the intra-day precision test ranged from 4.5 to 6.0% with respect to the concentration. Accuracy ranged from 1.0 to 4.0% (inter-day). The method was used for the determination of tramadol in a bioequivalence study. PMID- 9200540 TI - Simple and robust high-performance liquid chromatographic method for the determination of ranitidine in microvolumes of human serum. AB - A simple robust high-performance liquid chromatographic method is described for the determination of ranitidine in microvolumes of human serum. The drug of interest was isolated using liquid-liquid extraction with dichloromethane and back-extraction with 0.1% phosphoric acid and separation was obtained using a reversed-phase column under isocratic conditions, with ultraviolet detection at 313 nm. Intra-day and inter-day coefficients of variation ranged from 1 to 6% and 3 to 10%, respectively. Accuracy of the assay was less than 10% at all concentrations. The limit of detection and the limit of quantitation were 2 and 7 ng/ml, respectively. The linearity was assessed in the range 10-1000 ng/ml. It was shown that a group of common drugs co-administered with ranitidine did not interfere with its determination. The applicability of this method for the pharmacokinetic study of ranitidine following i.v. infusion in patients was demonstrated using only 100 microl of serum. The ruggedness of the assay was demonstrated over a three-year period. PMID- 9200541 TI - Determination of acyclovir in human plasma by high-performance liquid chromatography. AB - A selective and sensitive isocratic high-performance liquid chromatographic method for the analysis of acyclovir in human plasma was described. Acid deproteinisation was used as sample treatment. Mean analytical recoveries were higher than 94% at low and high concentrations. The quantification limit was 0.1 mg/l for a plasma volume of 500 microl and precision study exhibits coefficients of variation lower than 5%. The method is suitable for therapeutic monitoring of acyclovir concentrations in organ-transplant recipients. PMID- 9200542 TI - High-performance liquid chromatographic analysis of Peptide T in rabbit plasma with on-line column enrichment. AB - The present paper describes the development of a simple and sensitive analytical method for quantification of Peptide T (PT) in rabbit plasma, using standard analytical equipment and on-line column enrichment, without prior extraction, clean-up or derivatization. The analytical procedure was found to be accurate, precise and linear. The accuracy was 100% (range 97-103%) and the mean precision was 8% (range 3-14%) for all (n=6) concentrations (0, 15, 50, 100 and 200 ng/ml). The total recovery was found to be approximately 80%, and it was found to be dependent upon the injection rate onto the extraction column. The correlation between added and found concentrations was 0.9982, and the limit of detection was estimated to be around 5 ng/ml. The method is therefore found to be suitable for bioavailability studies, involving Peptide T, in rabbits. PMID- 9200543 TI - Simple high-performance liquid chromatographic method for the determination of acyclovir in human plasma using fluorescence detection. AB - A simple high-performance liquid chromatographic method using fluorescence detection was developed for the determination of acyclovir in human plasma. The method entailed direct injection of the plasma sample after deproteination. It is both specific and sensitive with a detection limit of 30 ng/ml at a signal-to noise ratio of 3:1, and is thus suitable for use in pharmacokinetic studies of acyclovir. The method had a mean absolute recovery of 96%, while the within-day and between-day coefficients of variation and percentages error were all less than 8%. The calibration curve was linear over a concentration range of 62.5-4000 ng/ml. PMID- 9200544 TI - Finasteride in biological fluids: extraction and separation by a graphitized carbon black cartridge and quantification by high-performance liquid chromatography. AB - A simple, specific and sensitive high-performance liquid chromatographic method has been developed and validated for the determination of finasteride in human plasma. A solid-phase extraction procedure was used to isolate finasteride from the biological matrix before quantitative analysis. The analyte was separated on a Symmetry reversed-phase column using acetonitrile-0.04 M orthophosphoric acid (pH 4.0) as mobile phase and quantified by measuring its UV absorbance at 215 nm. The limit of detection for the analyte was 0.005 microg/ml. 4-Androstene-3,17 dione was used as internal standard. The calibration graph of the method was linear from 0.01 to 3.0 microg/ml of finasteride in human plasma, and the coefficient of variation less than 4.5%. This HPLC procedure is simple, precise and accurate for the determination of finasteride in human plasma. PMID- 9200545 TI - Identification of Gingko biloba flavonol metabolites after oral administration to humans. AB - An extract of Ginkgo biloba leaves (EGb) was given to healthy volunteers. Urine samples were collected for 3 days, and blood samples were withdrawn every 30 min for 5 h. The samples were purified through SPE C18 cartridges and analyzed by reversed-phase LC-diode array detection for the presence of EGb metabolites. Only urine samples contained detectable amounts of substituted benzoic acids, i.e., 4 hydroxybenzoic acid conjugate, 4-hydroxyhippuric acid, 3-methoxy-4 hydroxyhippuric acid, 3,4-dihydroxybenzoic acid, 4-hydroxybenzoic acid, hippuric acid and 3-methoxy-4-hydroxybenzoic acid (vanillic acid). In contrast to rats no phenylacetic acid or phenylpropionic acid derivatives were found in urine, thus indicating that in humans a more extensive metabolism takes place. As for rats the metabolites found in human urines accounted for less than 30% of the flavonoids given. The same procedure was applied to blood samples, and no metabolites could be detected. PMID- 9200546 TI - Discriminative stimulus effects of the endogenous neuroactive steroid pregnanolone. AB - Naive male Sprague-Dawley rats were trained to discriminate the endogenous neuroactive steroid pregnanolone (5.6 mg/kg) from saline. Three positive modulators of the GABA(A) receptor complex substituted for pregnanolone: the neuroactive steroid allopregnanolone (1.0-10.0 mg/kg), the barbiturate pentobarbital (3.0-17.0 mg/kg), and the benzodiazepine diazepam (0.3-3.0 mg/kg). In contrast, buspirone, a 5-HT1A-mediated anxiolytic, failed to substitute up to rate-suppressing doses (1.0-5.6 mg/kg). The present experiment demonstrated the ability of an endogenous neuroactive steroid to function as a discriminative stimulus. Moreover, these results suggest that the discriminative stimulus effects of pregnanolone are mediated via positive modulation of GABA(A) receptors. PMID- 9200547 TI - Modulation by presynaptic adenosine A1 receptors of nicotinic receptor antagonist induced neuromuscular block in the mouse. AB - We have investigated how altering the activation of adenosine A1 receptors modifies nicotinic receptor antagonist-induced fade of tetanic contractions in the mouse isolated hemi-diaphragm. Vecuronium-induced tetanic fade was attenuated by an adenosine A1 receptor antagonist (8-cyclopentyl-1,3-dipropylxanthine, DPCPX, 10(-7) M) and by an inhibitor of the synthesis of extracellular adenosine from ATP (alpha,beta-methylene ADP, MeADP, 5 x 10(-5) M). Conversely, vecuronium induced tetanic fade was potentiated by an adenosine A1 receptor agonist (N6 cyclohexyladenosine, CHA, 10(-7) M) and an inhibitor of the extracellular destruction of adenosine (erythro-9-[2-hydroxy-3-nonyl]adenine, EHNA, 10(-4) M). The ability of an adenosine A1 receptor antagonist to attenuate vecuronium induced tetanic fade indicates that a component of this fade is due to endogenous adenosine. Further, the ability of the inhibitor of adenosine synthesis to attenuate vecuronium-induced tetanic fade indicates that this endogenous adenosine is derived from ATP. Hexamethonium-induced tetanic fade was also potentiated by increasing adenosine A1 receptor activation, albeit with a higher concentration of CHA (10(-4) M). However, unlike for vecuronium, hexamethonium induced tetanic fade was not attenuated by reducing adenosine A receptor activation. This latter observation suggests that the tetanic fade produced by hexamethonium and vecuronium does not share a common mechanism of action. PMID- 9200548 TI - Prolonged anticonvulsant action of glutamate metabotropic receptor agonists in inferior colliculus of genetically epilepsy-prone rats. AB - The anticonvulsant activity of (S)-4-carboxy-3-hydroxyphenylglycine ((S)-4C3HPG) (an antagonist of Group I and an agonist of Group II metabotropic glutamate (mGlu) receptors), of (1S,3S)-1-aminocyclopentane-1,3-dicarboxylic acid ((1S,3S) ACPD) (an agonist of Group II mGlu receptors), and of L-serine-O-phosphate (an agonist of Group III mGlu receptors) was studied against sound-induced seizures in genetically epilepsy-prone (GEP) rats following bilateral microinjection into the inferior colliculus. All 3 drugs produce dose-dependent suppression of all phases of sound-induced seizures (wild running, clonic and tonic). (S)-4C3HPG produces an immediate and short-lasting (< 2 h) protection against sound-induced seizures with an ED50 value of 4.3 (3.2-5.7) nmol, at 5 min. The preferential agonists of Group II and Group III mGlu receptors produce an immediate, transient (< 10 min) proconvulsant effect followed by a prolonged (> 1 day) anticonvulsant effect against sound-induced seizures. The anticonvulsant ED50 value for (1S,3S) ACPD is 9 (5-18) nmol at 2 h, and for L-serine-O-phosphate is 36 (6.5-199) nmol at 2 days. It is concluded that mGlu receptor activation potently modifies seizure threshold. PMID- 9200549 TI - Pharmacologically induced neural plasticity in the prefrontal cortex of adult gerbils (Meriones unguiculatus). AB - Using a selective antibody serum against glutaraldehyde-conjugated gamma aminobutyric acid (GABA), GABAergic neurons were identified in the medial prefrontal cortex of young adult gerbils (Meriones unguiculatus) following a single non-invasive dose of methamphetamine (25 mg/kg i.p.) applied at the age of 90 days. GABA-immunoreactive profiles were electron microscopically counted in a defined test field (0.875 mm2) covering the prefrontal prelimbic area after a single dose of either methamphetamine or saline. Within 30 days following the drug challenge the density of GABAergic innervation significantly increased by about 20%. Several lines of previous investigation indicate that a single dose of methamphetamine is an appropriate stimulus to cause selective autotoxic destruction of certain prefrontal dopamine fibres due to drug-induced hyperactivation. There is further indication of postsynaptic and transneuronal neuroplasticity since the densities of dendritic spines on prefrontal pyramidal cells went through a significant sequence of post-drug gain and loss. These structural dynamics resemble typical alterations seen after classical mechanical or chemical lesioning in other regions of the brain. The present results on drug induced reactive neuroplasticity are discussed together with the current understanding of stimulus-induced adaptive reorganization in the mammalian central nervous system. PMID- 9200550 TI - N6-2-(4-aminophenyl)ethyl-adenosine enhances the anticonvulsive activity of antiepileptic drugs. AB - N6-2-(4-Aminophenyl)ethyl-adenosine (APNEA, a non-selective agonist of the adenosine A3 receptors), at the subprotective dose of 1 mg/kg against electroconvulsions, significantly potentiated the anticonvulsive action of phenobarbital, diphenylhydantoin and valproate against maximal electroshock, being ineffective at lower doses. APNEA (0.0039-1 mg/kg) also enhanced the protective activity of carbamazepine. Aminophylline (5 mg/kg) and 8-cyclopentyl 1,3-dimethylxanthine (8-CPX, 5 mg/kg), reversed the APNEA (1 mg/kg)-induced enhancement of the anticonvulsive action of phenobarbital, diphenylhydantoin and valproate, but not that of carbamazepine produced by APNEA at 0.0039 mg/kg. The adenosine agonist did not alter the plasma levels of antiepileptic drugs studied, so a pharmacokinetic interaction is not probable. Finally, APNEA (0.0156 and 1 mg/kg) administered alone or in combination with carbamazepine significantly decreased the body temperature and impaired long-term memory. Our results suggest that APNEA at low doses potentiates the protective activity of carbamazepine most likely through the A subtype of adenosine receptors. At higher doses, APNEA seems to enhance the anticonvulsive effect of other antiepileptics via adenosine A1 receptors. PMID- 9200551 TI - Apamin-sensitive SK(Ca) channels modulate adrenal catecholamine release in anesthetized dogs. AB - We investigated the role of high conductance (BK(Ca)) and small conductance Ca2(+)-activated K+ (SK(Ca)) channels in adrenal catecholamine release in response to splanchnic nerve stimulation, acetylcholine, the nicotinic receptor stimulant 1,1-dimethyl-4-phenyl-piperazinium (DMPP), and muscarine in anesthetized dogs. The selective SK(Ca) channel blocker apamin and the selective BK(Ca) channel blocker charybdotoxin were infused into the adrenal gland through the phrenicoabdominal artery, and the cholinergic agonists were injected into the same artery. Splanchnic nerve stimulation (1, 2, 3 and 10 Hz), acetylcholine (0.75, 1.5 and 3 microg), DMPP (0.1, 0.2 and 0.4 microg) and muscarine (0.5, 1 and 2 microg) produced frequency- or dose-dependent increases in catecholamine output as measured in adrenal venous blood. Apamin infusion (1, 3 and 10 ng/min) enhanced the acetylcholine-, DMPP- and muscarine-induced increases in catecholamine output in a dose-dependent manner, but it did not affect the splanchnic nerve stimulation-induced catecholamine response. Charybdotoxin infusion (10, 30 and 100 ng/min) did not affect the increases in catecholamine output induced by the agonists and splanchnic nerve stimulation. Neither apamin nor charybdotoxin affected basal catecholamine output. These results suggest that apamin-sensitive SK(Ca) channels located in adrenal medullary cells may play an inhibitory role in the regulation of adrenal catecholamine release mediated by extrasynaptic nicotinic and muscarinic receptors. PMID- 9200552 TI - Estrogen augments cyclopiazonic acid-mediated, endothelium-dependent vasodilation. AB - The modulatory effects of chronic estrogen treatment on the responses to cyclopiazonic acid, an endoplasmic reticulum Ca2(+)-ATPase inhibitor, were studied in rings of aorta and the isolated perfused kidney of the rat. Rings of aorta were obtained from the following groups of age-matched rats (i) male, (ii) female, and two groups of rats implanted with a subcutaneous pellet (iii) ovariectomized, placebo-treated, (iv) ovariectomized, 17beta-estradiol-treated (0.5 mg/pellet/21 days). In phenylephrine (2 microM) pre-contracted rings with intact endothelium, cyclopiazonic acid (10(-7) to 3 x 10(-5) M) produced endothelium-dependent relaxations in a concentration-dependent manner. The cyclopiazonic acid dilation as a percentage loss of phenylephrine tone was greater in aortic rings from female (72.9 +/- 2.4%) and estrogen-treated rats (65.5 +/- 4.8%) compared to those from male (51.5 +/- 3.4%) or ovariectomized rats (40.8 +/- 3.9%) (P < 0.05, one-way analysis of variance (ANOVA)). These relaxation responses of cyclopiazonic acid were converted to contractions by pre treatment with an inhibitor of nitric oxide (NO) synthase, N(omega)-nitro-L arginine methyl ester (L-NAME, 200 microM; 30 min). There were no differences in cyclopiazonic acid-induced contractions of aortas excised from either estrogen treated or untreated-ovariectomized rats. In perfused kidneys, cyclopiazonic acid (10(-5) M) caused a larger decrease in perfusion pressure in kidneys from female rats (110 +/- 0.4 mmHg) than it did in kidneys from male rats (80 +/- 0.6 mmHg). These results demonstrate that cyclopiazonic acid causes a greater endothelium dependent dilation in estrogen-treated ovariectomized and control female rats, possibly due to unmasking of estrogen-enhanced Ca2+ entry into the endothelial cells. PMID- 9200553 TI - Alterations in alpha subunit expression of cardiac Na+,K+-ATPase in spontaneously hypertensive rats: effect of antihypertensive therapy. AB - The alpha-2 subunit abundance of Na+,K(+)-ATPase in the rat heart has been reported to be reduced in several induced hypertensive models. To determine whether this reduction also occurs in a genetic model of hypertension, we studied expression of the alpha subunits in left ventricles of spontaneously hypertensive rats (SHR), and normotensive Wistar-Kyoto (WKY) and Sprague-Dawley rats using Western blotting and quantitative dot-blotting analysis with monoclonal antibodies. While the alpha-1 subunit was not affected in any of the strains, a significant reduction of the alpha-2 subunit expression was noted in 19-week-old SHRs, but not in age-matched WKY and Sprague-Dawley rats, supporting the hypothesis that elevated arterial pressure may differentially downregulate the alpha-2 subunit in the rat heart. To further test this hypothesis we designed experiments in which hypertensive rats were treated with the antihypertensive agents hydralazine and nifedipine. Both agents effectively normalized the blood pressure in the SHRs with no significant effect on the blood pressure in the WKY and Sprague-Dawley rats. The alpha-2 subunit in SHRs treated with hydralazine and nifedipine showed a 63.3% (n = 6, P < 0.05, analysis of variance and Fischer's test) and a 27.4% increase, respectively, over the hypertensive SHR controls, although the reversal effect of nifedipine did not quite reach significance. The alpha-1 subunit expression was not affected by any of the drug treatments. No effect of either of the drugs on the alpha-1 or alpha-2 subunit was observed in the WKY or Sprague-Dawley rat groups. These data support our hypothesis that the alpha-2 subunit may be a pressure-sensitive isoform of the cardiac Na+,K(+) ATPase and that high blood pressure is, directly or indirectly, responsible for the reduction of the alpha-2 subunit protein expression. PMID- 9200554 TI - Failure of heparin to inhibit the expression of the thrombin receptor following endothelial injury of the rabbit carotid artery. AB - The effect of heparin on thrombin receptor expression was evaluated in an experimental model of myointimal smooth muscle cell proliferation in rabbits. Myointimal hyperplasia was induced by an air-drying injury of the carotid artery and thrombin receptor expression following endothelial injury was measured by in situ hybridisation and immunohistochemistry. In healthy arteries, thrombin receptor mRNA and protein were detected in the endothelial cells only. In contrast, 14 days after endothelial injury, thrombin receptor mRNA expression increased in the smooth muscle cells present in the neointima, predominantly in areas of active cell proliferation. A 2-week subcutaneous treatment with heparin (10 mg/kg per day, s.c.) inhibited smooth muscle cell hyperplasia occurring in the intima following deendothelialization (80 +/- 7.8% inhibition, P < 0.001). The 14-day heparin treatment strongly reduced thrombin receptor gene and protein expression observed in the endothelial cells in healthy arteries but did not affect thrombin receptor expression which occurred in smooth muscle cells which have proliferated in the neointima as a consequence of endothelial injury. These results therefore establish that thrombin receptor expression during intimal hyperplasia is an heparin-insensitive event. PMID- 9200555 TI - The dog saphenous vein: a sensitive and selective preparation for the Y2 receptor of neuropeptide Y. AB - The dog saphenous vein responds to neuropeptide Y with a dose-dependent contraction and this vasopressor effect is not altered by the removal of the endothelium nor by the neuropeptide Y Y1 receptor antagonist, BIBP 3226 ((R)-N2 (diphenylacetyl)-N-[(n-hydroxyphenyl)methyl]-argininami de). The dose-response curves obtained with neuropeptide Y, peptide YY and with C-terminal fragments such as neuropeptide Y-(2-26), neuropeptide Y-(13-36) and peptide YY-(3-36) have similar slopes and maxima. EC50 values of these compounds vary between 30 +/- 10 and 89 +/- 47 nM. The neuropeptide Y Y1 receptor-selective agonist [Leu31,Pro34]neuropeptide Y and human pancreatic polypeptide are inactive up to 1 microM. This pharmacological profile suggests that the contraction of the dog saphenous vein induced by neuropeptide Y and its homologues or fragments is mediated by a neuropeptide Y Y2 receptor type. Moreover, this neuropeptide Y Y2 receptor appears to be localized in the venous smooth muscle, where it exerts a direct myotropic effect that may be useful for the pharmacological characterization of new compounds acting as agonists or antagonists of the neuropeptide Y Y2 receptor. PMID- 9200556 TI - FR167653, a dual inhibitor of interleukin-1 and tumor necrosis factor-alpha, ameliorates endotoxin-induced shock. AB - Increased production of interleukin-1 and tumor necrosis factor-alpha (TNF-alpha) have been implicated in the pathophysiology of a variety of diseases including circulatory shock. The present study evaluated the efficacy of FR167653 (1-[7-(4 fluorophenyl)-1,2,3,4-tetrahydro-8-pyridylpyrazolo[5,1-c] [1,2,4]triazin-2-yl]-2 phenylethanedione sulfate monohydrate), a dual inhibitor of interleukin-1 and TNF alpha production, to protect rabbits from the shock and lethality induced by lipopolysaccharide. In this sepsis model, FR167653 at a dose of 0.32 mg/kg per h ameliorated the 7-day mortality from 93% in the placebo group to 47% in the FR167653-treated group and, at doses of 0.10-0.32 mg/kg per h, attenuated the hypotensive response to lipopolysaccharide challenge and returned mean arterial blood pressure to almost normal levels. The increases in plasma interleukin-1 and TNF-alpha levels evoked by lipopolysaccharide administration were also inhibited by treatment with FR167653, which was efficacious at doses of 0.1-0.32 mg/kg per h. In addition, FR167653 treatment attenuated the increases in plasma creatinine concentrations consistent with renal damage in a dose-dependent manner. These findings suggested that FR 167653 has a beneficial potential as a drug for septic shock or multiple organ dysfunction syndrome. PMID- 9200557 TI - Inhibitory effects of a fullerene derivative, dimalonic acid C60, on nitric oxide induced relaxation of rabbit aorta. AB - Dimalonic acid C60 (10(-5) M), a new fullerene derivative, produced an augmentation of phenylephrine-induced tone and reduced both the acetylcholine induced maximum relaxation and the amplitude of substance P (10(-8) M)-induced relaxation in endothelium-containing thoracic aorta of rabbit; the acetylcholine- and substance P-induced relaxation was restored in the presence of superoxide dismutase (250 U/ml). Dimalonic acid C60 (10(-5) M) did not influence the phenylephrine-induced contractile response in the absence of endothelium, but the acetylcholine-induced relaxation was eliminated by removal of the endothelium. Superoxide anion generation, using hypoxanthine (1 mM)/xanthine oxidase (16 mU/ml), reduced the acetylcholine-induced relaxation and produced an augmentation of phenylephrine-induced tone in endothelium-containing strips; these effects were negated by the addition of superoxide dismutase (250 U/ml). A nitric oxide generating agent, S-nitroso-N-acetylpenicillamine, caused relaxation of aorta without endothelium in a concentration-dependent manner, and the concentration response curve was shifted to the right in the presence of dimalonic acid C60. This inhibitory effect of dimalonic acid C60 was also masked in the presence of superoxide dismutase. Sodium nitroprusside-induced relaxation was not affected by either dimalonic acid C60 or superoxide dismutase. These observations suggest that dimalonic acid C60 inhibits endothelium (nitric oxide)-dependent agonist induced relaxation through the production of superoxide. PMID- 9200558 TI - Effects of endothelin-1 on arterial and venous resistances in anaesthetized rats. AB - The effects of endothelin-1 and vehicle (0.9% NaCl) on mean arterial pressure, heart rate, mean circulatory filling pressure, systemic arterial resistance, cardiac output and venous resistance were studied in four groups of pentobarbitone-anaesthetized rats, either in presence or absence of phentolamine. I.v. bolus injections of endothelin-1 at 0.5, 1 and 2 nmol/kg dose dependently increased mean arterial pressure (22, 34 and 40 mmHg), arterial resistance (33, 93 and 122% over baseline), venous resistance (40, 117 and 143% over baseline) and mean circulatory filling pressure (1.0, 1.7 and 1.8 mmHg), but decreased heart rate (-16, -21 and -17 beats/min) and cardiac output (-6, -28 and -35% below baseline). The vehicle did not significantly alter any of these variables. During the continuous infusion of phentolamine (300 microg/kg per min), endothelin-1 caused similar increases in arterial resistance, venous resistance and mean circulatory filling pressure, similar reduction in cardiac output but significantly greater pressor and bradycardic responses, suggesting that the arterial and venous constrictor effects of endothelin-1 are not due to sympathetic activation and the stimulation of alpha-adrenoceptors. The results show that endothelin-1 raised mean arterial pressure via the increment in systemic arterial resistance, since cardiac output was markedly reduced. This decrease in cardiac output was mediated by increases in arterial as well as venous resistances. The vasoconstrictor and venoconstrictor effects of endothelin 1 were independent of sympathetic tone. PMID- 9200559 TI - Effect of a selective 5-HT3 receptor agonist on gastric motility in fasted and fed dogs. AB - The effect of m-chlorophenylbiguanide, a selective 5-HT3 receptor agonist, on gastric antral motility was investigated in conscious dogs with a force transducer implanted chronically. m-Chlorophenylbiguanide (0.1-1 mg/kg i.v.) dose dependently enhanced antral motility in the fasted state, and the amplitude of m chlorophenylbiguanide (1 mg/kg i.v.)-induced antral contractions reached the level of natural phase III contractions. In contrast, m-chlorophenylbiguanide reduced the amplitude of antral contractions in the fed state. A selective 5-HT3 receptor antagonist, ramosetron (0.0003-0.03 mg/kg i.v.), inhibited both effects of m-chlorophenylbiguanide. m-Chlorophenylbiguanide (1 mg/kg i.v.)-induced contractions were inhibited by atropine (0.03 or 0.1 mg/kg i.v.). These results indicate that pharmacological activation of 5-HT3 receptors has opposite effects on canine gastric antral motility in the fasted and in the fed state, being stimulatory and inhibitory, respectively. The stimulatory effect seems to be mediated mainly via the release of acetylcholine. PMID- 9200560 TI - Tolterodine--a new bladder-selective antimuscarinic agent. AB - Tolterodine is a new muscarinic receptor antagonist intended for the treatment of urinary urge incontinence and other symptoms related to an overactive bladder. The aim of the present study was to compare the antimuscarinic properties of tolterodine with those of oxybutynin, in vitro and in vivo. Tolterodine effectively inhibited carbachol-induced contractions of isolated strips of urinary bladder from guinea pigs (K(B) 3.0 nM; pA2 8.6; Schild slope 0.97) and humans (K(B) 4.0 nM; pA2 8.4; Schild slope 1.04) in a concentration-dependent, competitive manner. The affinity of tolterodine was similar to that derived for oxybutynin (K(B) 4.4 nM; pA2 8.5; Schild slope 0.89) in the guinea-pig bladder. Tolterodine (21-2103 nmol/kg (0.01-1 mg/kg); intravenous infusion) was significantly more potent in inhibiting acetylcholine-induced urinary bladder contraction than electrically-induced salivation in the anaesthetised cat. In contrast, oxybutynin displayed the opposite tissue selectivity. Radioligand binding data showed that tolterodine bound with high affinity to muscarinic receptors in urinary bladder (K(i) 2.7 nM), heart (K(i) 1.6 nM), cerebral cortex (K(i) 0.75 nM) and parotid gland (K(i) 4.8 nM) from guinea pigs and in urinary bladder from humans (K(i) 3.3 nM). Tolterodine and oxybutynin were equipotent, except in the parotid gland, where oxybutynin bound with 8-times higher affinity (K(i) 0.62 nM). Binding data on human muscarinic m1-m5 receptors expressed in Chinese hamster ovary cells showed that oxybutynin, in contrast to tolterodine, exhibits selectivity (10-fold) for muscarinic m3 over m2 receptors. The K(B) value determined for oxybutynin (4.4 nM) in functional studies on guinea-pig bladder correlated better with the binding affinity at muscarinic M2/m2 receptors (K(i) 2.8 and 6.7 nM) than at muscarinic M3/m3 receptors (K(i) 0.62 and 0.67 nM). The tissue selectivity demonstrated for tolterodine in vivo cannot be attributed to selectivity for a single muscarinic receptor subtype. However, the combined in vitro and in vivo data on tolterodine and oxybutynin may indicate either that muscarinic M3/m3 receptors in glands are more sensitive to blockade than those in bladder smooth muscle, or that muscarinic M2/m2 receptors contribute to bladder contraction. PMID- 9200561 TI - Involvement of ATP-sensitive K+ channels in the inhibitory effect of calcitonin gene-related peptide on neurotransmission in rat vas deferens. AB - This study investigated the inhibitory action of human calcitonin-gene-related peptide (CGRP) on neurotransmission in rat isolated vas deferens. The electrically stimulated contractile responses, which were mediated predominantly by activation of postganglionic noradrenergic nerve fibers, were concentration dependently inhibited by human CGRP (0.1-100 nM, IC50 = 2.15 +/- 0.21 nM, n = 17). Human CGRP at concentrations greater than 3 nM reduced the contractile responses to exogenous noradrenaline and ATP. The inhibitory effect of human CGRP on the electrically stimulated or agonist-induced contractions was antagonized by human CGRP-(8-37), the CGRP receptor antagonist. Glibenclamide (3-10 microM) decreased the effect of human CGRP at a concentration greater than 1 nM whilst glibenclamide did not affect the inhibitory effect of human CGRP on the agonist induced contractions. These results indicate that human CGRP at low concentrations exerts its inhibitory action mainly by acting on CGRP receptors at the sympathetic nerve terminals supplying rat vas deferens and the activation of ATP-sensitive K+ channels is at least in part involved in the action of human CGRP on neurotransmission. PMID- 9200562 TI - (-)-Deprenyl treatment restores serum insulin-like growth factor-I (IGF-I) levels in aged rats to young rat level. AB - We studied the effects of treatment with (-)-deprenyl, a monoamine oxidase B inhibitor, on plasma levels of insulin-like growth factor-I (IGF-I) (as indicator of growth hormone (GH) secretion), levels of monoamines and their metabolites, and the activity and content of tyrosine hydroxylase - the rate-limiting enzyme in the biosynthesis of catecholamines - in the hypothalamus and hypophysis of old male rats. Male Wistar rats (22 months old) were treated with 2 mg deprenyl/kg body weight s.c. three times a week for 2 months. At the end of the treatment period, blood was collected for measurement of plasma IGF-I levels by radioimmunoassay (RIA). The concentrations of dopamine, serotonin (5-HT) and their main metabolites were determined by high performance liquid chromatography (HPLC) with electrochemical detection, and the tyrosine hydroxylase content in hypothalamus and hypophysis was determined by enzyme-linked immunoabsorbent assay (ELISA). (-)-Deprenyl treatment produced a pronounced increase in dopamine and 5 HT in both the hypothalamus and hypophysis (P < 0.01). The main dopaminergic metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC), decreased in hypothalamus but not in hypophysis, and treatment had no effect on the concentration of 5 hydroxyindole-3-acetic acid (5-HIAA). The tyrosine hydroxylase activity and tyrosine hydroxylase content increased in hypothalamus and hypophysis (P < 0.05). In the hypophysis the increase in tyrosine hydroxylase activity was consistent with the increase in tyrosine hydroxylase amount. Moreover, (-)-deprenyl treatment restored the IGF-I plasma levels in old rats to a concentration similar to those found in young animals. Postulated anti-aging effects of (-)-deprenyl could hence be due to restoration of hypothalamic hormones such as GH. PMID- 9200563 TI - A selective inhibitor of cyclooxygenase-2 reverses endotoxin-induced pyretic responses in non-human primates. AB - The anti-pyretic effect of a selective cyclooxygenase-2 inhibitor, DFU (5,5 dimethyl-3-(3-fluorophenyl)-4-(4-methylsulfonyl)phenyl-2(5H)-furano ne), was examined in conscious, un-restrained squirrel monkeys (Saimiri sciureus) using a radio telemetric system. Injection of bacterial endotoxin (lipopolysaccharide, 6 microg kg(-1), i.v.) in squirrel monkeys caused a gradual increase in core body temperature reaching a plateau of 2.07 +/- 0.17 degrees C above baseline at 2 h post-injection. Oral administration of DFU (1 mg kg(-1)) reduced, and DFU (3 mg kg(-1)) completely reversed the lipopolysaccharide-induced pyretic responses. The onset of action of DFU (about 30 min) is in good agreement with the pharmacokinetic profile of this compound in squirrel monkeys. The effect of DFU is comparable to that of a conventional non-selective non-steroidal anti inflammatory drug (NSAID), diclofenac (3 mg kg(-1)). Since the plasma levels achieved for DFU at the dose employed in the present study are below the threshold required for inhibition of cyclooxygenase-1, it is concluded that the anti-pyretic effect of DFU can be attributed predominantly to an inhibitory action on cyclooxygenase-2. Thus, lipopolysaccharide-induced pyresis in squirrel monkeys can be used as a model for evaluation of anti-pyretic activity of cyclooxygenase inhibitors. PMID- 9200564 TI - Expression of a brain-type cannabinoid receptor (CB1) in alveolar Type II cells in the lung: regulation by hydrocortisone. AB - Using the polymerase chain reaction with degenerate primers to identify novel G protein-coupled receptors of the rat alveolar Type II cell, we identified sequences expressed by the Type II cell identical to the sequence of the rat brain cannabinoid receptor (CB1). The use of Northern blot analysis to examine expression of CB1 mRNA in rat tissues revealed differences between the brain and lung. While rat brain expressed a 6.0 kb mRNA as previously described, rat lung expressed mRNA of 4.5 and 6.0 kb. Isolated lung alveolar Type II cells also expressed mRNA of 4.5 and 6.0 kb as determined by Northern analysis. However, only freshly isolated Type II cells contained cannabinoid receptor mRNA. Reverse transcriptase-polymerase chain reaction (RT-PCR) failed to detect CB1 mRNA in Type II cells maintained in culture for 1 or 2 days. We next determined developmental changes in lung CB1 mRNA expression using semi-quantitative RT-PCR. CB1 expression was detected as early as gestational day 16 in rat lung and mRNA levels increased to fetal day 20 before birth, before declining to adult levels. Fetal rat lung explants were utilized to further examine the ontogeny and hormonal effects on CB1 mRNA expression. Hydrocortisone induced a dose-dependent expression in 15-day and 18-day explants, similar to previous results for surfactant-associated proteins. Our results demonstrate expression of CB1 mRNA in rat alveolar Type II cells and rat lung. This expression is ontogenically and hormonally regulated, with maximal expression noted just prior to birth in rat lung. Since CB1 mRNA is only expressed in freshly isolated Type II cells, CB1 may be useful as a Type II cell marker. PMID- 9200566 TI - Overcoming of multidrug resistance by VA-033, a novel derivative of apovincaminic acid ester. AB - We have studied the effects of a novel derivative of apovincaminic acid ester, VA 033, on the resistance of tumors to chemotherapeutic agents. VA-033 increased the sensitivity of drug-resistant cell lines (P388/VCR, P388/ADM, AD10, and K562/ADM) to adriamycin or vincristine. The potency of VA-033 was stronger than verapamil. The drug lengthened the survival time of the P388/VCR-implanted mice treated with vincristine. VA-033 increased the intracellular accumulation of vincristine in the tumor cells, and the photolabeling of P-glycoprotein by [3H]azidopine was inhibited by VA-033. VA-033 showed a slight inhibitory effect on the L-type Ca2+ current in the ventricular myocytes, and had less effect on the cardiovascular parameters such as blood pressure, contractile force and atrio-ventricular conduction time than verapamil when administered systemically in the dog. These results suggest that VA-033 may become a beneficial compound as a modifier to the neoplastic cell resistant to multidrugs. PMID- 9200565 TI - Ionic mechanisms of tetrandrine in cultured rat aortic smooth muscle cells. AB - The ionic mechanism of tetrandrine, an alkaloid extracted from the Chinese medicinal herb Radix stephania tetrandrae, was investigated in A7r5 vascular smooth muscle cells. The nystatin-perforated whole-cell voltage-clamp technique was performed to examine the effects of tetrandrine on ionic currents. Tetrandrine (1-100 microM) reversibly caused an inhibition of L-type voltage dependent Ca2+ current (I(Ca,L)) in a concentration-dependent manner. Tetrandrine did not cause any change in the overall shape of the current-voltage relationship of I(Ca,L). The IC50 value of tetrandrine-induced inhibition of I(Ca,L) was 5 microM. In the presence of Bay K 8644 (3 microM) or cyclopiazonic acid (30 microM), tetrandrine still produced a significant inhibition of I(Ca,L). The inhibitory effects of tetrandrine on I(Ca,L) exhibited tonic and use-dependent characteristics. Moreover. tetrandrine (3 microM) shifted the steady-state inactivation curve of I(Ca,L) to more negative membrane potentials by approximately -15 mV. These results indicate that tetrandrine directly inhibits the voltage-dependent L-type Ca2+ current in vascular smooth muscle cells, which may predominantly contribute to the vasodilatory actions of tetrandrine. PMID- 9200567 TI - Binding profile of the novel 5-HT1B/1D receptor antagonist, [3H]GR 125,743, in guinea-pig brain: a comparison with [3H]5-carboxamidotryptamine. AB - Native brain 5-HT1B/1D) receptors were studied using the novel antagonist, [3H]GR 125,743 (N-[4-methoxy-3-(4-methylpiperazin-1-yl)phenyl]-3-methyl-4-(4-pyri dyl)benzamide). In guinea-pig striatal membranes, [3H]GR 125,743 displayed rapid association (t1/2 = 4.5 min), high (90%) specific binding and high affinity (K(d) = 0.29 nM), although B(max) values (fmol/mg protein) varied according to brain region-striatum: 199; frontal cortex: 89; hippocampus: 79; cerebellum: 26. In frontal cortex, the B(max) determined with [3H]5-CT ([3H]carboxamidotryptamine) was significantly higher (178; P < 0.05), suggesting that it also labels other binding sites. In striatal membranes, guanylylimidodiphosphate (GppNHp) inhibited [3H]5-CT but not [3H]GR 125,743 binding, suggesting that the latter has antagonist properties. Nevertheless, in competition binding experiments, the pK(i) values obtained with [3H]GR 125,743 and [3H]5-CT for 20 serotonergic ligands, including L 694,247 (2-[5-[3-(4-methylsulphonylamino)benzyl-1,2,4 oxadiazol-5-yl ]-1H-indole-3-yl]ethylamine), GR46,611 (3-[3-(2-dimethylamino ethyl)-1H-indol-6-yl]-N-(4-methoxybenzyl)acrylami de), sumatriptan and alniditan, were highly correlated (r = 0.99). Ketanserin and ritanserin showed low affinity for [3H]GR 125,743 binding to guinea-pig striatal sites (K(i) = 12600 and 369 nM), suggesting that 5-HT1B (rather than 5-HT1D) receptors are predominantly labelled in this tissue. The present data indicate that [3H]GR 125,743 is a useful tool for studying native 5-HT1B/1D receptors. PMID- 9200568 TI - Tetracaine stimulates extracellular Ca2+-independent insulin release. AB - The effect of the local anesthetic, tetracaine, on 45Ca efflux, cytoplasmic Ca2+ concentration [Ca2+]i and insulin secretion in pancreatic B-cells was studied. At a physiological level of [Ca2+]o, tetracaine (0.1-5 mM) dose-dependently inhibited insulin secretion induced by 22 mM glucose. Paradoxically, at the same glucose concentration but in the absence of external Ca2+, tetracaine dose dependently increased insulin secretion. At a low glucose level (2.8 mM) tetracaine failed to affect secretion, either in the presence or absence of external Ca2+. At high (22 mM) or low (2.8 mM) glucose, [Ca2+]i was increased by tetracaine in a dose-dependent manner. Tetracaine (2 mM) also increased the 45Ca efflux from isolated islets. This effect was of the same magnitude at both low and high glucose concentrations, and was independent of the presence of extracellular Ca2+. Finally, tetracaine increased 45Ca efflux from islets perifused in the presence of thapsigargin. In conclusion, our data indicate that tetracaine releases Ca2+ from a thapsigargin-insensitive store in pancreatic B cells. Under suitable experimental conditions, insulin release can be elicited by a [Ca2+]o-independent pathway. The existence of a ryanodine-like Ca2+ channel in pancreatic B-cells is proposed. PMID- 9200569 TI - Factors influencing learners' specialty decisions. PMID- 9200570 TI - Another strategy for rescuing teaching hospitals. PMID- 9200571 TI - Concern about new California licensure requirement. PMID- 9200572 TI - Making optimal use of information technology. PMID- 9200573 TI - The unrecognized medical educator. PMID- 9200574 TI - Improving relationships with commercial supporters of medical education. PMID- 9200575 TI - Reflections of the past in today's applicant pool. PMID- 9200576 TI - Institutional challenges posed by faculty development. PMID- 9200577 TI - Reflections on the scholarship of teaching. PMID- 9200578 TI - How one teaching hospital system and one medical school are jointly affirming their academic mission. AB - The economic forces that are reshaping the practice of medicine and the funding of medical research will have great impact on clinical education and research in teaching hospitals and their associated medical schools. Changes in the setting of and approach to medical education will need to be made in order to continue to train physicians at the same high level as in the past and to maintain the productivity of our national biomedical research enterprise and its contributions to health. Academic leaders, such as department chiefs who have clinical service responsibilities, are finding it more and more difficult to manage simultaneously the demands of the clinical business, education, and research. In an effort to organize a teaching hospital and a medical school in a manner that would position them to maintain more effectively their common academic mission front and center with the clinical business, Harvard Medical School and the Beth Israel Hospital created a joint venture in 1996. The new nonprofit Institute for Education and Research has education and research as its top (and only) mission. It is designed to provide additional and specific academic leadership and to enable the joint venture to undertake strategic planning for the academic mission. In addition to the challenges it faces from changes in the external environment, the Institute for Education and Research will need to establish a new pattern of interactions internally within the parent institutions. Collaborations with department chairs and faculty are an essential ingredient for its success. It is hoped that this structure will prove to be a useful template for organizing other medical school hospital collaborations on behalf of the academic mission. PMID- 9200579 TI - Beyond corporate-style downsizing: a better way for medical schools to succeed in a changing world. AB - There is a critical need for medical schools and universities to consider strategies beyond corporate-style downsizing to address revenue needs and reposition their organizations. The author presents considerable evidence and three reasons to reject downsizing as a way to facilitate long-term organizational success. Instead, she recommends that institutions use a comprehensive approach to individual and organizational development to assure a flexible, enduring organization. Specifically, medical schools and universities should take an institution-wide perspective and approach to continually training, retraining, or reassigning faculty and should continually adapt their organizational structures and procedures as necessary to achieve changing institutional goals. The result will be the retention of able and dedicated faculty, who will be crucial in helping their schools continue to be successful while adapting to a changing world. PMID- 9200580 TI - Implementing a comprehensive approach to managing faculty roles, rewards, and development in an era of change. AB - The current environment in which medicine is taught and practiced requires that medical schools pay increased attention to the faculty member's roles, rewards, career development, and productivity. Medical schools must make strategic decisions about the allocation of resources that can nurture their faculties and support the activities in academic and community settings in which faculty are involved. From 1993 to 1995 Allegheny University of the Health Sciences (formerly Medical College of Pennsylvania and Hahnemann University) designed a comprehensive system for the professional development of faculty. This system is based upon expanded categories of faculty academic activity and scholarship. New programs were implemented to reorient faculty toward conducting and documenting the expanded array of scholarly activities. The main characteristics of the new system are the establishment of formally defined performance expectations, the vertical alignment of the individual faculty member's objectives with the department's mission and the school's mission, and an increasing emphasis upon faculty interdependence, accountability, and use of sound business practices. The authors describe these and other aspects of the design of the new system in detail and report initial results and lessons learned from the system's implementation, evaluation, and dissemination throughout the university. The long term success of this comprehensive professional development program will be assessed over time by observing how this institution advances its mission in a well-planned and cost-effective manner that retains talented, productive, and professionally fulfilled faculty. PMID- 9200581 TI - Ideas for medical education. Introduction. PMID- 9200582 TI - Defining and evaluating quality for ambulatory care educational programs. AB - As the training of medical students and residents increasingly moves to ambulatory care settings, clerkship and program directors must find a way to use their limited resources to guide the development and evaluation of the quality of these ambulatory-based learning experiences. To evaluate quality, directors must first define, in operational and measurable terms, what is meant by the term "quality" as it is applied to ambulatory-based education. Using educational theories and the definition of quality used by health care systems, the authors propose an operational definition of quality for guiding the planning, implementation, and evaluation of ambulatory care educational programs. They assert that quality is achieved through the interaction of an optimal learning environment, defined educational goals and positive outcomes, participant satisfaction, and cost-effectiveness. By describing the components of quality along with examples of measurable indicators, the authors provide a foundation for the evaluation and improvement of instructional innovations in ambulatory care education for the benefit of teachers, learners, and patients. PMID- 9200583 TI - Perspectives on computing and medical education. Introduction. PMID- 9200584 TI - "Just-in-time" clinical information. AB - The just-in-time (JIT) model originated in the manufacturing industry as a way to manage parts inventories process so that specific components could be made available at the appropriate times (that is, "just in time"). This JIT model can be applied to the management of clinical information inventories, so that clinicians can have more immediate access to the most current and relevant information at the time they most need it--when making clinical care decisions. The authors discuss traditional modes of managing clinical information, and then describe how a new, JIT model may be developed and implemented. They describe three modes of clinician-information interactions that a JIT model might employ, the scope of information that may be made available in a JIT model (global information or local, case-specific information), and the challenges posed by the implementation of such an information-access model. Finally, they discuss how JIT information access may change how physicians practice medicine, various ways JIT information may be delivered, and concerns about the trustworthiness of electronically published and accessed information resources. PMID- 9200585 TI - Medicine and the arts. Charlotte's Web. PMID- 9200586 TI - Financing academic medicine: strengthening the tangled strands before they snap. PMID- 9200587 TI - Improving the privacy and security of electronic health information. PMID- 9200588 TI - A systematic analysis of how medical school characteristics relate to graduates' choices of primary care specialties. AB - PURPOSE: To examine medical school characteristics, in particular federal funding for biomedical research, as they relate to the graduates' choices of family medicine, general internal medicine, general pediatrics, or all three specialties. METHOD: Data were collected for 121 U.S. medical schools, including information on funding, faculty, curricula, and other school characteristics. In addition, a questionnaire was mailed to the schools requesting information about non-federal funding for primary care, primary care department characteristics, and primary care representation on the admission, curriculum, and promotion and tenure committees. Analyses were carried out separately for each specialty and for all three combined. The first multiple regression analysis was done to predict specialty choice (proximate predictors), the second to predict the predictors of specialty choice (intermediate predictors), and the third to predict those predictors (distal predictors). RESULTS: Prediction was best for family medicine practice. Interest at matriculation and required third-year and fourth-year time in primary care were the two best proximate predictors. The best predictors of initial interest were the percentage of rural students and special programs for primary care, while the best predictors of required time in primary care were funding for family medicine and the percentage of faculty in family medicine (intermediate predictors). The best predictor of the percentage of faculty in family medicine was funding for family medicine (distal predictor). CONCLUSION: The results suggest that the most effective way to increase the number of physicians with generalist practices is to increase the number of students interested in a family medicine career at matriculation. PMID- 9200589 TI - A study of medical students' specialty-choice pathways: trying on possible selves. AB - PURPOSE: To describe the decision-making processes reported by graduating medical students in choosing primary care (PC) or non-primary-care (NPC) specialties. METHOD: Members of the University of Washington School of Medicine's graduating class of 1995 were invited to participate in focus groups. Six specialty-choice pathways were defined based on a previously administered survey of recalled preferences at matriculation and stated choice at the time of the National Resident Matching Program. Students were assigned to focus groups based on specialty-choice pathway. Transcribed discussions and summaries were thematically coded and analyzed using grounded theory and quantitative comparisons. RESULTS: Of 157 students, 140 (89%) completed the initial survey, and 133 (85%) provided enough information to be classified by pathway. In all, 47 students participated in the focus group discussions. The PC students cited PC orientation, diversity of patients and activities, role models and mentors, interaction with patients, and overall medical school culture as having influenced their choice. The NPC students cited lifestyle, controllable hours, opportunities to do procedures, therapeutic urgency and effect, active tempo, exciting settings, and intellectual challenge. Role models influenced PC career choice much more than NPC career choice, and often served to refute negative stereotypes. The sense of personal fit between themselves and specialties was important to the students in all groups, but differed in emphasis according to career-choice pathways. Those whose preferences did not change experienced a confirmation of pre-existing beliefs, while those who switched specialty areas developed a sense of fit through the inclusion or elimination of different practice aspects. Those who switched specialty areas reported more negative influences and misunderstanding of their initially preferred specialties. CONCLUSION: The process of specialty choice can be described usefully as a socially constructed process of "trying on possible selves" (i.e., projecting oneself into hypothetical career and personal roles). This may explain role models' exceptional influence in disproving negative stereotypes. Medical students' choices can best be facilitated by recognizing their needs to gain knowledge not only about specialty content, but also about practitioners' lives and the students' own present and possible selves. PMID- 9200591 TI - Costs of preceptors' time spent teaching during a third-year family medicine outpatient rotation. AB - PURPOSE: To quantify the cost of teaching a student in a family physician's office in terms of the time a preceptor spends teaching or of the possible decrease in a preceptor's productivity, as reflected in a smaller number of patients seen per day. METHOD: During July and August 1995 data were collected from 26 different preceptor-student pairs (the third-year students were attending the University of Cincinnati College of Medicine). A single research assistant was used to time the actions of the students and the preceptors and to record every activity (under one of four categories) of the preceptors that related to the students. Also recorded were the numbers of patients seen by the preceptor with and without the student and, if the preceptor was in a partnership or group practice, the number of patients seen by the preceptor's partner on the same day in the same office. RESULTS: For ten preceptors with partners, there was no significant difference between the number of patients seen by the preceptor and the number seen by a non-teaching partner. The preceptors averaged 0.17 hours listening to the students' presentations of patients, 0.51 hours waiting for the students to finish seeing a patient, 2.55 hours seeing patients with the students in the examination room, and 0.56 hours giving mini-lectures or testing the students' knowledge, for a total of 3.79 hours per day in student-related activities. CONCLUSION: Much of the 3.79 hours the preceptors spent with the students was spent seeing patients who would have been seen even if the preceptors had not been teaching. The best estimate of "extra" time spent by the preceptors would be the time spent lecturing or testing plus the time spent listening to presentations plus an estimated 20% of the time spent seeing patients with the students, a total of 1.23 hours. At $60 per hour, the cost of extra preceptor time would be $73.80 per day, or $1,254.60 per student for a typical four-week rotation. PMID- 9200590 TI - A longitudinal study of students' depression at one medical school. AB - PURPOSE: Using a standardized measure of depression at three assessment points, to examine depression in medical students during their training. METHOD: Students entering the University of Massachusetts Medical School in the fall in 1987, 1988, and 1989 were mailed a recruitment letter and baseline questionnaire four weeks prior to the start of classes. Subsequent assessments took place in the middles of year 2 and year 4 and included only the students who had participated in the baseline assessment. The baseline assessment included the Center for Epidemiological Studies Depression (CES-D) scale, the Bortner Type A Behavior scale, the Spielberger Trait Anger scale, and the Spielberger Anger Expression scale. In addition, the baseline package included a rating of perceived stress, a demographics questionnaire, and a social-life survey. The follow-up assessments included the CES-D scale, the rating of perceived stress level, and the social life survey. Analytic methods used were univariate descriptive statistics, correlation, and multiple-linear-regression analyses, two-sample t-tests, analysis of variance, and chi-square tests. RESULTS: Of the initial pool of 300 students, 264 responded at the baseline assessment (88% response rate; 53% men); 171 of these participated in the year-2 assessment (65% response rate; 51% men), and 126 participated in the year-4 assessment (48% response rate; 48% men); a total of 99 students participated in all three assessments. CES-D scores > or =80th percentile were obtained for 18% of the entering students. This rose to 39% at year 2 and 31% at year 4 (p = .0001). No gender difference was found at baseline; however, the women experienced higher depression levels than did the men at year 2 (p = .004) and at year 4 (p = .04). Overall, gender and increases in perceived stress (from baseline to year 2) were significant predictors of increased CES-D scores (from baseline to year 2; p = .01 and p = .0001, respectively). For the women, increased perceived stress, angerin, and frequency of social contacts outside work/school were significant predictors of the magnitude of increases in CES-D scores (baseline to year 2; p = .0001, p = .02, and p = .03, respectively). CONCLUSION: These preliminary data support the view that, upon entering medical school, students' emotional status resembles that of the general population. However, the rise in depression scores and their persistence over time suggest that emotional distress during medical school is chronic and persistent rather than episodic. Also, the women had more significant increases in depression scores than did the men. PMID- 9200592 TI - Measuring the effects of problem-based learning on the development of veterinary students' clinical expertise. AB - PURPOSE: To investigate whether repeated exposure to simulated clinical cases, as employed in problem-based learning (PBL), accelerates the development of clinical expertise in veterinary students. METHOD: In 1995 all 122 second-year students at the Texas A&M University College of Veterinary Medicine were organized into 24 self-selected groups to complete four computer-based case simulations in veterinary neuroanatomy. A scoring rubric of four categories (poor, neutral, good, and excellent) was used to assign quality classifications for physical examination questions selected by the students. Each group's diagnostic efficiency was calculated for each case on the basis of these classifications. An analysis of variance (ANOVA) using a one-factor repeated-measures procedure was employed to examine the cumulative effects of the case simulations on diagnostic efficiency. Post hoc procedures involved the use of contrasts to determine the trend of diagnostic efficiency with repeated use of case simulations. A Pearson product-moment correlation coefficient was calculated to determine the strength of the relationship between the change in diagnostic efficiency for each group and the selected group characteristics, total hands-on veterinary experience, and average grade-point average (GPA). RESULTS: Because six of the 24 groups did not perform a physical examination in one of the cases, a rating of zero was given when no physical examination was done; the data were then analyzed with and without the six groups included, and compared. The ANOVA yielded a significant result for all 24 groups (F(3,69) = 2.75, p = .0491) and for the 18 groups that performed physical examinations in all four cases (F(3,51) = 3.03, p = .0377). Significant linear contrasts were also found for all 24 groups (F(1,69) = 7.21, p = .009) and for the 18 groups (F(1,51) = 4.25, p = .044). Improvement in diagnostic efficiency could not be correlated with GPAs and was only somewhat correlated with the total amount of prior clinical experience reported by the students. CONCLUSION: Study findings suggest that there is a significant relationship between the repeated use of case simulations in PBL and the accelerated development of clinical expertise. PMID- 9200593 TI - Introducing case management to a general medicine ward team of a teaching hospital. AB - PURPOSE: To introduce case management to a general medicine ward team of a teaching hospital to improve patient care and ensure comprehensive longitudinal care. METHOD: The Department of Veterans Affairs Medical Center is one of four hospitals used by University of Oklahoma School of Medicine residents. There are five medicine teams, each comprising a second- or third-year resident, one or two interns, two medical students, and a faculty physician. The case-management program was initiated in November 1994. No attempt was made to limit the residents assigned to the case-managed team (i.e., many residents who worked with the case-managed team subsequently rotated through the other teams). Patients were assigned to the teams by rotation, and no attempt was made to adjust for the severity of illness among admissions. The teams were separated as follows: pre case-management teams (all five teams prior to the case-management program), non case-management teams (the four teams without case managers after the program's initiation), and the case-management team. The study periods were January-July 1994 (pre-case management) and January-July 1995 (after case management). RESULTS: The numbers of patients treated by the three groups were 1,305, 1,139, and 289, respectively. The median length of stay for pre-case-management patients was 5 days (interquartile range, 3-9 days); for non-case-management patients, 5 days (range, 3-8 days); and for case-management patients, 5 days (range, 3-7 days). The cumulative distribution of lengths of stay for case-management patients was significantly different from those of the other study groups by the Kolmogorov-Smirnov test (p = .02). More case-management patients were discharged by day 7. Rates of readmission were not significantly different between the teams. CONCLUSION: In this study a case-management program was effectively implemented in a teaching hospital, resulting in reduced lengths of stay for patients. As academic health centers become more concerned with efficiency and cost, case management should be seriously considered as a way to deal with such issues. PMID- 9200594 TI - A study of peer-review marking reveals weaknesses. PMID- 9200595 TI - A faculty-development needs assessment at one medical school. PMID- 9200596 TI - A preliminary study of students' comfort and preparedness with different types of patient groups. PMID- 9200599 TI - The on and off of floral regulatory genes. PMID- 9200597 TI - Results of the National Resident Matching Program for 1997. PMID- 9200598 TI - Tuberculosis--search for the cure. PMID- 9200600 TI - Microtubule function in morphological differentiation: growth zones and growth cones. PMID- 9200601 TI - Starting at the beginning, middle, and end: translation initiation in eukaryotes. PMID- 9200602 TI - Properties of H. volcanii tRNA intron endonuclease reveal a relationship between the archaeal and eucaryal tRNA intron processing systems. AB - To better understand the relationship between archaeal and eucaryal tRNA introns and their processing systems, we have cloned the gene encoding the tRNA intron endonucleases from the archaeon H. volcanii. The gene encodes a 37 kDa protein that appears to be present as a homodimer under native conditions. Recombinant forms of this protein were expressed in E. coli and found to cleave precursor tRNAs lacking full mature tRNA structure, a property observed for the native endonuclease. Comparative sequence analysis revealed that similar proteins existed in other Archaea and that these proteins have significant similarity with two subunits of the yeast tRNA intron endonuclease. These results provide evidence that the archaeal and eucaryal tRNA intron processing systems are related and suggest a common origin for tRNA introns in these organisms. PMID- 9200603 TI - The yeast tRNA splicing endonuclease: a tetrameric enzyme with two active site subunits homologous to the archaeal tRNA endonucleases. AB - The splicing of tRNA precursors is essential for the production of mature tRNA in organisms from all major phyla. In yeast, the tRNA splicing endonuclease is responsible for identification and cleavage of the splice sites in pre-tRNA. We have cloned the genes encoding all four protein subunits of endonuclease. Each gene is essential. Two subunits, Sen2p and Sen34p, contain a homologous domain of approximately 130 amino acids. This domain is found in the gene encoding the archaeal tRNA splicing endonuclease of H. volcanii and in other Archaea. Our results demonstrate that the eucaryal tRNA splicing endonuclease contains two functionally independent active sites for cleavage of the 5' and 3' splice sites, encoded by SEN2 and SEN34, respectively. The presence of endonuclease in Eucarya and Archaea suggests an ancient origin for the tRNA splicing reaction. PMID- 9200604 TI - The eucaryal tRNA splicing endonuclease recognizes a tripartite set of RNA elements. AB - The tRNA splicing endonuclease cleaves intron-containing tRNA precursors on both sides of the intron. The prevailing belief has been that the enzyme binds only to the mature domain through the invariant bases. We show instead that, for recognition, the endonuclease utilizes distinct sets of structural elements, several of which are within the intron. One subset of recognition elements, localized in the mature domain, is needed for recognition of both cleavage sites, while two other subsets, localized at the exon-intron boundaries, are used for recognition of either one or the other cleavage site. The two cleavage sites are essentially independent: neither is required by the other for cleavage to take place. These results support a two-active-site model for the eucaryal endonuclease. PMID- 9200605 TI - An RNA 5'-triphosphatase related to the protein tyrosine phosphatases. AB - mRNA capping requires the sequential action of three enzymatic activities: RNA triphosphatase, guanylyl-transferase, and methyltransferase. Here we characterize a gene (CEL-1) believed to encode the C. elegans capping enzyme. CEL-1 has a C terminal domain containing motifs found in yeast and vaccinia virus capping enzyme guanylyltransferases. The N-terminal domain of CEL-1 has RNA triphosphatase activity. Surprisingly, this domain does not resemble the vaccinia virus capping enzyme but does have significant sequence similarity to the protein tyrosine phosphatase (PTP) enzyme family. However, CEL-1 has no detectable PTP activity. The mechanism of the RNA triphosphatase is similar to that of PTPs: the active site contains a conserved nucleophilic cysteine required for activity. These results broaden the superfamily of PTP-like phosphatases to include enzymes with RNA substrates. PMID- 9200606 TI - Structural and functional analysis of the mitotic rotamase Pin1 suggests substrate recognition is phosphorylation dependent. AB - The human rotamase or peptidyl-prolyl cis-trans isomerase Pin1 is a conserved mitotic regulator essential for the G2/M transition of the eukaryotic cell cycle. We report the 1.35 A crystal structure of Pin1 complexed with an AlaPro dipeptide and the initial characterization of Pin1's functional properties. The crystallographic structure as well as pH titration studies and mutagenesis of an active site cysteine suggest a catalytic mechanism that includes general acid base and covalent catalysis during peptide bond isomerization. Pin1 displays a preference for an acidic residue N-terminal to the isomerized proline bond due to interaction of this acidic side chain with a basic cluster. This raises the possibility of phosphorylation-mediated control of Pin1-substrate interactions in cell cycle regulation. PMID- 9200607 TI - Structure of an enzyme required for aminoglycoside antibiotic resistance reveals homology to eukaryotic protein kinases. AB - Bacterial resistance to aminoglycoside antibiotics is almost exclusively accomplished through either phosphorylation, adenylylation, or acetylation of the antibacterial agent. The aminoglycoside kinase, APH(3')-IIIa, catalyzes the phosphorylation of a broad spectrum of aminoglycoside antibiotics. The crystal structure of this enzyme complexed with ADP was determined at 2.2 A. resolution. The three-dimensional fold of APH(3')-IIIa reveals a striking similarity to eukaryotic protein kinases despite a virtually complete lack of sequence homology. Nearly half of the APH(3')-IIIa sequence adopts a conformation identical to that seen in these kinases. Substantial differences are found in the location and conformation of residues presumably responsible for second-substrate specificity. These results indicate that APH(3') enzymes and eukaryotic-type protein kinases share a common ancestor. PMID- 9200608 TI - Frpo: a novel single-stranded DNA promoter for transcription and for primer RNA synthesis of DNA replication. AB - We describe a novel promoter for E. coli RNA polymerase that functions efficiently only in the form of single-stranded DNA. Derived from the leading region of F plasmid, single-stranded Frpo sequence directs RNA polymerase to initiate transcription at a specific site within Frpo, and this specific transcription is highly stimulated by SSB. Prior denaturation activates transcription from otherwise inactive duplex DNA containing Frpo. Since RNAs synthesized on SSB-coated single-stranded Frpo are efficiently elongated into DNA chains by DNA polymerase III holoenzyme, transcription at Frpo serves also for priming DNA replication. A mode of recognition by RNA polymerase of a unique secondary structure within Frpo is proposed, and possible roles of this novel single-stranded promoter in expression and replication during conjugal transfer of F plasmid are discussed. PMID- 9200609 TI - SH2 signaling in a lower eukaryote: a STAT protein that regulates stalk cell differentiation in dictyostelium. AB - The TTGA-binding factor is a transcriptional regulator activated by DIF, the chlorinated hexaphenone that induces prestalk cell differentiation in Dictyostelium. The same activity also functions as a repressor, controlling stalk cell differentiation. We show that the TTGA-binding factor is a STAT protein. Like the metazoan STATs, it functions via the reciprocal interaction of a phosphotyrosine residue on one molecule with an SH2 domain on a dimerizing partner. Furthermore, it will bind specifically to a mammalian interferon stimulated response element. In Saccharomyces cerevisiae, where the entire genomic sequence is known, SH2 domains have not been identified. It would seem, therefore, that SH2 signaling pathways arose very early in the evolution of multicellular organisms, perhaps to facilitate intercellular comunication. PMID- 9200610 TI - An exported peptide functions intracellularly to contribute to cell density signaling in B. subtilis. AB - Competence development and sporulation in B. subtilis are partly controlled by peptides that accumulate in culture medium as cells grow to high density. We constructed two genes that encode mature forms of two different signaling molecules, the PhrA peptide that stimulates sporulation, and CSF, the competence- and sporulation-stimulating factor. Both pentapeptides are normally produced by secretion and processing of precursor molecules. The mature pentapeptides were functional when expressed inside the cell, indicating that they normally need to be imported to function. Furthermore, at physiological concentrations (10 nM), CSF was transported into the cell by the oligopeptide permease encoded by spo0K (opp). CSF was shown to have at least three different targets corresponding to its three activities: stimulating competence gene expression at low concentrations, and inhibiting competence gene expression and stimulating sporulation at high concentrations. PMID- 9200611 TI - Chaperonin-mediated folding in the eukaryotic cytosol proceeds through rounds of release of native and nonnative forms. AB - The eukaryotic cytosolic chaperonin, CCT, plays an essential role in mediating ATP-dependent folding of actin and tubulin. There is debate about whether it mediates folding through a single round of association followed by release of native forms, or through cycles of binding and full release in which only a fraction of released molecules reaches native form in any cycle. We examine the fate of newly synthesized substrate proteins bound to CCT in reticulocyte lysate or intact Xenopus oocytes. When a chaperonin "trap," able to bind but not release substrate protein, is introduced, production of the native state is strongly inhibited, associated with transfer to trap. While predominantly nonnative forms of actin, tubulin, and a newly identified substrate, G(alpha)-transducin, are released from CCT, a small fraction reaches native form with each round of release, inaccessible to trap. This overall mechanism resembles that of the bacterial chaperonin, GroEL. PMID- 9200612 TI - tea1 and the microtubular cytoskeleton are important for generating global spatial order within the fission yeast cell. AB - Fission yeast cells identify and maintain growing regions exactly opposed at the ends of a cylindrical cell. tea1 mutants disrupt this organization, producing bent and T-shaped cells. We have cloned tea1 and shown that tea1 is located at the cell poles. Microtubules are continuously required to transfer tea1 to the cell ends, and tea1 is located at the ends of microtubules growing toward the cell poles. We suggest that tea1 acts as an end marker, directing the growth machinery to the cell poles. tea1 is down-regulated in cells treated with pheromone that grow toward a mating partner and no longer maintain their ends exactly opposed. tea1 may also influence microtubular organization, affecting the maintenance of a single central axis. PMID- 9200613 TI - Cocrystal structure of the messenger RNA 5' cap-binding protein (eIF4E) bound to 7-methyl-GDP. AB - The X-ray structure of the eukaryotic translation initiation factor 4E (eIF4E), bound to 7-methyl-GDP, has been determined at 2.2 A resolution. eIF4E recognizes 5' 7-methyl-G(5')ppp(5')N mRNA caps during the rate-limiting initiation step of translation. The protein resembles a cupped hand and consists of a curved, 8 stranded antiparallel beta sheet, backed by three long alpha helices. 7-methyl GDP binds in a narrow cap-binding slot on the molecule's concave surface, where 7 methyl-guanine recognition is mediated by base sandwiching between two conserved tryptophans, plus formation of three hydrogen bonds and a van der Waals contact between its N7-methyl group and a third conserved tryptophan. The convex dorsal surface of the molecule displays a phylogenetically conserved hydrophobic/acidic portion, which may interact with other translation initiation factors and regulatory proteins. PMID- 9200614 TI - Calcium-induced restructuring of nuclear envelope and endoplasmic reticulum calcium stores. AB - The spatial organization of endoplasmic reticulum (ER) and nuclear envelope (NE) calcium stores is important for the regulation of localized calcium signals and sustained calcium gradients. Here, we have used a lumenal GFP fusion protein and shown that, in resting cells, large molecules can rapidly diffuse across the cell within the lumenal storage space defined by the ER and NE membranes. Increases in cytosolic calcium concentration reversibly fragmented ER tubules and prevented lumenal diffusion. However, the integrity of the NE was maintained, and a significant fraction of NE lumenal protein accumulated in an NE-associated vesicle. These dynamic properties of ER-NE calcium stores provide insights into the spatiotemporal control of calcium signaling. PMID- 9200615 TI - The store-operated calcium current I(CRAC): nonlinear activation by InsP3 and dissociation from calcium release. AB - Patch-clamp experiments aimed at determining the relationship between intracellular Ca2+ release and activation of store-operated calcium current I(CRAC) reveal that both agonist and InsP3-mediated activation of I(CRAC) are highly nonlinear, occurring over a narrow concentration range. Ca2+ release and Ca2+ influx can be dissociated, as they possess differential sensitivities to InsP3: low concentrations induce substantial Ca2+ release without any activation of I(CRAC), whereas micromolar concentrations of InsP3 are required to activate Ca2+ influx. This suggests functionally distinct stores controlling Ca2+ release and influx and enables cells to switch between sources of Ca2+ to fit best their current needs. PMID- 9200617 TI - A piece of my mind. Bea's legacy. PMID- 9200616 TI - A requirement for Flk1 in primitive and definitive hematopoiesis and vasculogenesis. AB - Mouse embryos lacking the receptor tyrosine kinase, Flk1, die without mature endothelial and hematopoietic cells. To investigate the role of Flk1 during vasculogenesis and hematopoiesis, we examined the developmental potential of Flk1 /- embryonic stem cells in chimeras. We show that Flk1 is required cell autonomously for endothelial development. Furthermore, Flk1-/- cells do not contribute to primitive hematopoiesis in chimeric yolk sacs or definitive hematopoiesis in adult chimeras and chimeric fetal livers. We also demonstrate that cells lacking Flk1 are unable to reach the correct location to form blood islands, suggesting that Flk1 is involved in the movement of cells from the posterior primitive streak to the yolk sac and, possibly, to the intraembryonic sites of early hematopoiesis. PMID- 9200618 TI - When US medicine became imperial. PMID- 9200619 TI - Lessons from US history of drug use. PMID- 9200620 TI - Screening to help asthmatics breathe easier. PMID- 9200621 TI - Military makes 'house calls' in 100 countries. PMID- 9200622 TI - Ronna Siegel, MD, is 17th Fishbein Fellow. PMID- 9200623 TI - From the Centers for Disease Control and Prevention. Transmission of nosocomial Legionnaires disease. PMID- 9200624 TI - Glucocorticoid use and risks of ocular hypertension and glaucoma. PMID- 9200625 TI - Glucocorticoid use and risks of ocular hypertension and glaucoma. PMID- 9200626 TI - Glucocorticoid use and risks of ocular hypertension and glaucoma. PMID- 9200627 TI - Elevated cerebrospinal fluid glutamate in patients with HIV-related dementia. PMID- 9200628 TI - The negative side of cost-effectiveness analysis. PMID- 9200629 TI - Modifiable risk factors for pulmonary embolism. PMID- 9200630 TI - Predicting progression of Alzheimer disease. PMID- 9200631 TI - Is managed care really here to stay? PMID- 9200632 TI - Is managed care really here to stay? PMID- 9200633 TI - Implementation of the Ottawa ankle rules in France. A multicenter randomized controlled trial. AB - OBJECTIVES: To assess the impact of the implementation of the Ottawa ankle rules on radiography requests in French hospitals during a 5-month intervention period and the impact of using posters alone to sustain the effect of the rules during a 5-month postintervention period. DESIGN: Multicenter randomized controlled trial preceded and followed by observational studies of radiological practices. SETTING: The emergency departments of 5 Paris university teaching hospitals of the Assistance Publique-Hopitaux de Paris. PATIENTS: A total of 2218, 1911, and 851 patients-all aged 18 years and older-who were seen for acute ankle or midfoot injuries in emergency departments during preintervention, intervention, and postintervention periods, respectively. INTERVENTION: Implementation of the Ottawa ankle rules by emergency department physicians in the intervention hospitals (using meetings, posters, pocket cards, and data forms). During the postintervention period, posters alone were used to sustain the intervention effect. MAIN OUTCOME MEASURE: Percentage of patients for whom radiography was requested. RESULTS: During the preintervention period, 98% and 98.5% of patients were referred for radiography in the intervention and control groups, respectively. During the intervention period, the mean proportions of patients referred for radiography by physicians was 78.9% in the intervention group and 99% in the control group (P=.03). Between preintervention and intervention periods, a relative reduction of 22.4% (95% confidence interval [CI], 19.8% 24.9%) in radiography requests was observed in the intervention group, while requests increased by 0.5% (95% CI, 0%-1.4%) in the control group. During the postintervention period, the proportion of radiography requests in the intervention hospitals was lower than the proportion observed in the preintervention period (83.1% vs 98%). CONCLUSIONS: Implementation of the Ottawa ankle rules significantly reduced radiography requests in French hospitals. Using a minimal postintervention implementation strategy, the effect of this intervention decreased but persisted after it was discontinued. PMID- 9200634 TI - Social ties and susceptibility to the common cold. AB - OBJECTIVE: To examine the hypothesis that diverse ties to friends, family, work, and community are associated with increased host resistance to infection. DESIGN: After reporting the extent of participation in 12 types of social ties (eg, spouse, parent, friend, workmate, member of social group), subjects were given nasal drops containing 1 of 2 rhinoviruses and monitored for the development of a common cold. SETTING: Quarantine. PARTICIPANTS: A total of 276 healthy volunteers, aged 18 to 55 years, neither seropositive for human immunodeficiency virus nor pregnant. OUTCOME MEASURES: Colds (illness in the presence of a verified infection), mucus production, mucociliary clearance function, and amount of viral replication. RESULTS: In response to both viruses, those with more types of social ties were less susceptible to common colds, produced less mucus, were more effective in ciliary clearance of their nasal passages, and shed less virus. These relationships were unaltered by statistical controls for prechallenge virus specific antibody, virus type, age, sex, season, body mass index, education, and race. Susceptibility to colds decreased in a dose-response manner with increased diversity of the social network. There was an adjusted relative risk of 4.2 comparing persons with fewest (1 to 3) to those with most (6 or more) types of social ties. Although smoking, poor sleep quality, alcohol abstinence, low dietary intake of vitamin C, elevated catecholamine levels, and being introverted were all associated with greater susceptibility to colds, they could only partially account for the relation between social network diversity and incidence of colds. CONCLUSIONS: More diverse social networks were associated with greater resistance to upper respiratory illness. PMID- 9200635 TI - Dose-related efficacy of levomethadyl acetate for treatment of opioid dependence. A randomized clinical trial. AB - OBJECTIVE: To compare the clinical efficacy of different doses of levomethadyl acetate hydrochloride (known as LAAM) in the treatment of opioid dependence. DESIGN: A randomized controlled, double-blind, parallel group, 17-week study. SETTING: Outpatient facilities at Johns Hopkins University Bayview Medical Center, Baltimore, Md. PATIENTS: Opioid-dependent volunteers (N=180) applying to a treatment-research clinic. INTERVENTION: Thrice-weekly (Monday/Wednesday/Friday) oral LAAM dose conditions of 25/25/35 mg, 50/50/70 mg, and 100/100/140 mg and nonmandatory counseling. MAIN OUTCOME MEASURES: Retention in treatment, self-reported heroin use, and opioid-positive urine specimens. RESULTS: Retention was independent of subjects' sex and dose. Self-reported heroin use decreased in a dose-related manner. At final assessment, patients in the high-dose condition reported using heroin 2.5 of 30 days as compared with 4.1 or 6.3 days for patients in the medium-dose and low-dose conditions, respectively (high dose vs low dose, P<.05); urinalysis results were similarly dose related. Overall, 20 (34%) of 59 patients in the high-dose condition remained opioid abstinent for 4 consecutive weeks, as compared with 8 (14%) of 59 in the medium dose and 7 (11%) of 62 in the low-dose conditions (P<.01). Self-report and urinalysis data are consistent with a greater than 90% reduction in illicit opioid use by the high-dose group relative to pretreatment levels. CONCLUSION: Opioid substitution treatment with LAAM substantially reduces illicit opioid use. The clinical efficacy of LAAM is positively related to dose. PMID- 9200636 TI - Trends in importation of measles to the United States, 1986-1994. AB - OBJECTIVES: To describe patterns among imported measles cases to the United States. DESIGN: Descriptive analysis of national case-based surveillance data on measles cases. SETTING: United States in the period 1986 through 1994. PATIENTS: All reported confirmed cases of measles. MAIN OUTCOME MEASURES: Demographic variables, immunization history, country of exposure, and reporting state. RESULTS: The number of reported imported cases of measles to the United States has dropped from an average of 99 cases annually in 1986 through 1988 and 190 cases in 1989 through 1991 to 61 cases in 1992 through 1994. Since 1990, the number of imported cases originating in Latin America declined by 98%, despite continued increase in the number of travelers to this region; cases from other regions remained relatively constant. This decrease paralleled the rapid decrease in measles incidence in the Western Hemisphere associated with national measles elimination programs. Most imported cases occurred among children, although 22% of cases occurred among young adults. Rates of measles cases per 1 million travelers are higher among non-US citizens than among US citizens. CONCLUSIONS: The sharp decline in importations into the United States from Latin America since 1991 provides evidence of the success of measles control efforts undertaken there. The decrease in imported cases has been associated with a decline in total measles cases in the United States. Sustained elimination of measles in the United States will require improved measles control in other countries in addition to a high level of population immunity. PMID- 9200637 TI - Pulmonary function in space. AB - The lung is exquisitely sensitive to gravity, and so it is of interest to know how its function is altered in the weightlessness of space. Studies on National Aeronautics and Space Administration (NASA) Spacelabs during the last 4 years have provided the first comprehensive data on the extensive changes in pulmonary function that occur in sustained microgravity. Measurements of pulmonary function were made on astronauts during space shuttle flights lasting 9 and 14 days and were compared with extensive ground-based measurements before and after the flights. Compared with preflight measurements, cardiac output increased by 18% during space flight, and stroke volume increased by 46%. Paradoxically, the increase in stroke volume occurred in the face of reductions in central venous pressure and circulating blood volume. Diffusing capacity increased by 28%, and the increase in the diffusing capacity of the alveolar membrane was unexpectedly large based on findings in normal gravity. The change in the alveolar membrane may reflect the effects of uniform filling of the pulmonary capillary bed. Distributions of blood flow and ventilation throughout the lung were more uniform in space, but some unevenness remained, indicating the importance of nongravitational factors. A surprising finding was that airway closing volume was approximately the same in microgravity and in normal gravity, emphasizing the importance of mechanical properties of the airways in determining whether they close. Residual volume was unexpectedly reduced by 18% in microgravity, possibly because of uniform alveolar expansion. The findings indicate that pulmonary function is greatly altered in microgravity, but none of the changes observed so far will apparently limit long-term space flight. In addition, the data help to clarify how gravity affects pulmonary function in the normal gravity environment on Earth. PMID- 9200638 TI - Antiretroviral therapy for HIV infection in 1997. Updated recommendations of the International AIDS Society-USA panel. AB - OBJECTIVE: To provide current recommendations for antiretroviral therapy for human immunodeficiency virus (HIV) disease. PARTICIPANTS: The original International AIDS Society-USA 13-member panel representing international expertise in antiretroviral research and care of patients with HIV infection. EVIDENCE: The following were considered: Newly available clinical and basic science study results, including phase 3 controlled trials; clinical, virological, and immunologic end-point data; interim analyses of studies presented at national and international research conferences; studies of HIV pathophysiology; and expert opinions of panel members. Recommendations were limited to the drugs available in mid 1997. PROCESS: The full panel met on a regular basis (July 1996, September 1996, November 1996, January 1997, and April 1997) since the publication of its initial recommendations in mid 1996 to review new research reports and interim results. The panel discussed whether and how new information changed its initial recommendations. The recommendations contained herein were determined by group consensus. CONCLUSIONS: New data have provided a stronger rationale for earlier initiation of more aggressive therapy than previously recommended and reinforce the importance of careful selection of initial drug regimen for each patient for optimal long-term clinical benefit and adherence. The plasma viral load is a crucial element of clinical management for assessing prognosis and the effectiveness of therapy, and such testing must be done properly. Treatment failure is most readily indicated by a rising plasma HIV RNA level and should be confirmed prior to a change of treatment. Therapeutic approaches must be updated as new data, particularly on the long-term clinical effect of aggressive antiretroviral treatment, continue to emerge. PMID- 9200639 TI - A 36-year-old woman recuperating from a stroke. PMID- 9200640 TI - A 17-year-old mother seeking contraception, 1 year later. PMID- 9200641 TI - Practice guidelines and prediction rules should be subject to careful clinical testing. PMID- 9200642 TI - Augmentation of hepatic glucose uptake by a positive glucose gradient between hepatoportal and central nervous systems. AB - To determine the role of the glucose gradient between the hepatoportal system (HPS) and the central nervous system (CNS) in regulating hepatic glucose uptake, experiments were conducted with seven conscious dogs using a hepatic venous catheterization technique. With the infusion of somatostatin (0.8 microg x kg(-1) x min(-1)), glucagon (0.65 ng x kg(-1) x min(-1)), and insulin (27 pmol x kg(-1) x min(-1)), arterial glucose levels could be maintained at 8 mmol/l by adjusting the intravenous glucose infusion (G(inf)) according to the following three periods: 1) peripheral glucose infusion period (PE), G(inf) alone; 2) portal glucose infusion period (PO), G(inf) plus constant glucose infusion into the portal vein (GIR(PV), 55.6 micromol x kg(-1) x min(-1)); 3) portal and brain glucose infusion period (PO+CNS), G(inf) and GIR(PV) plus additional glucose infusion into the unilateral carotid and vertebral arteries to abolish the positive glucose gradient between HPS and CNS. Arterial plasma glucose levels were clamped during the three periods (8.1 +/- 0.1, PE; 8.2 +/- 0.1, PO; 8.2 +/- 0.1 mmol/l, PO+CNS). During PO, when a positive glucose gradient was promoted between HPS and CNS, the net hepatic glucose balance (NHGB) determined by the difference between hepatic glucose inflow and outflow was significantly lower than that of PE (-41.5 +/- 5.3, PO vs. -7.5 +/- 3.4 micromol x kg(-1) x min(-1), PE; P < 0.01). However, this decrease in the NHGB significantly increased during PO+CNS, when the glucose gradient between HPS and CNS was minimized, compared with PO (-21.7 +/- 3.2 micromol x kg(-1) x min(-1), P < 0.05). We conclude that a positive glucose gradient between HPS and CNS is an important regulatory factor of hepatic glucose uptake, but other factors also play important roles because minimizing the glucose gradient between HPS and CNS diminished the net hepatic glucose uptake by 50%. PMID- 9200643 TI - Incorporation of [3-3H]glucose and 2-[1-14C]deoxyglucose into glycogen in heart and skeletal muscle in vivo: implications for the quantitation of tissue glucose uptake. AB - 2-deoxyglucose has been widely used to quantitate tissue glucose uptake in vivo, assuming that 2-deoxyglucose is transported and phosphorylated but not further metabolized. We examined the validity of this assumption by infusing [3 3H]glucose and 2-[1-14C]deoxyglucose in a similar primed continuous fashion to chronically catheterized, freely moving rats during normoglycemic hyperinsulinemic conditions. The rates of 2-deoxyglucose uptake were determined from the accumulation of 2-[1-14C]deoxyglucose-6-phosphate and 2-[1 14C]deoxyglucose-6-phosphate combined with the rate of the incorporation of 2-[1 14C]deoxyglucose into glycogen in rectus abdominis muscle and the heart. When the rates of glycogen synthesis during the 2-h hyperinsulinemic period from the two tracers were compared in rectus abdominis muscle, the rate of glycogen synthesis was twofold higher when measured with [3-3H]glucose (337 +/- 14 micromol x kg(-1) x min(-1)) than when measured with 2-[1-14C]deoxyglucose (166 +/- 10 micromol x kg(-1) x min(-1), P < 0.001). In the heart, the rate of glycogen synthesis was twofold higher when measured with 2-[1-14C]deoxyglucose (141 +/- 20 micromol x kg(-1) x min(-1)) than when measured with [3-3H]glucose (72 +/- 15 micromol x kg( 1) x min(-1), P < 0.001). The rate of 2-deoxyglucose uptake was 29% underestimated in rectus abdominis muscle, when counts found in glycogen were not included in glucose uptake calculations (398 +/- 25 vs. 564 +/- 25 micromol x kg( 1) x min(-1), P < 0.001). In the heart, glucose uptake was underestimated by 7% if glycogen counts were not taken into account (1,786 +/- 278 vs. 1,926 +/- 291 micromol x kg(-1) dry x min(-1), P < 0.05). The fraction of [3-3H]glucose incorporated into glycogen of total glucose metabolism (calculated from 2 deoxyglucose conversion to 2-deoxyglucose-6-phosphate and glycogen) was 0.6 (337/564) in rectus abdominis muscle and 0.037 (72/1,926) in the heart. We conclude that 2-deoxyglucose is incorporated into glycogen in the heart and in skeletal muscle in vivo under normoglycemic hyperinsulinemic conditions in the rat. Failure to consider the incorporation of 2-deoxyglucose into glycogen will underestimate the rate of tissue glucose uptake. To avoid such problems, the amount of 2-deoxyglucose incorporated into glycogen should be quantitated in subsequent studies. PMID- 9200644 TI - Fatty acids mediate the acute extrahepatic effects of insulin on hepatic glucose production in humans. AB - We have shown previously in humans that insulin partly suppresses hepatic glucose production (HGP) by an extrahepatic (indirect) mechanism. In the present study, we investigated the role of free fatty acids (FFAs) in mediating the extrahepatic effects of insulin in humans and determined the extent to which insulin can regulate HGP by a non-FFA-mediated effect. Sixteen healthy men received an intravenous tolbutamide infusion for 3 h, and pancreatic insulin secretion was calculated by deconvolution of peripheral C-peptide levels. On a subsequent occasion, equimolar exogenous insulin was infused by peripheral vein. In both studies, glucose was clamped at euglycemia. We have previously validated this method and shown no independent insulin-like activity of tolbutamide. During the clamp, 9 of the 16 subjects received a low dose of heparin and Intralipid to prevent the insulin-induced suppression of FFAs, while 7 subjects received a high dose of heparin and Intralipid to raise FFAs approximately 2.5-fold. In both the high- and low-dose groups, peripheral insulin was higher and calculated portal insulin lower with peripheral versus portal insulin delivery. In the low-dose group, HGP decreased by 68.3 +/- 2.1% with portal insulin delivery and 64.7 +/- 3.7% with peripheral insulin delivery (NS). In the high-dose group, HGP decreased by 58.0 +/- 4.5% with portal insulin and 48.3 +/- 5.0% with peripheral insulin (P < 0.05). Four individuals who participated in the high-dose group underwent an additional peripheral insulin study in which the same dose of exogenous insulin was infused as in the high-dose group but in the absence of heparin and Intralipid. During this latter study, FFA levels declined by approximately 90% during hyperinsulinemia, and HGP was suppressed by 71.8 +/- 5.6%, which was a much greater suppression (P < 0.01) than when FFA levels were raised in these subjects during the equivalent rate insulin infusion. In summary, the previously observed greater suppression of HGP with equimolar peripheral versus portal insulin is eliminated or reversed, depending on plasma FFA levels, if FFAs are prevented from decreasing, suggesting an important role of FFAs in mediating the extrahepatic effects of insulin on HGP. However, the effect of FFA clamping is relatively small with a significant degree of suppression of HGP (by approximately 50%), which remains even when FFAs are elevated above basal levels, suggesting that in the physiological range FFAs only partially influence the suppression of HGP in humans. This suggests that other mechanisms, most likely hepatic, dominate the acute insulin-induced suppression of glucose production. PMID- 9200645 TI - Improved human islet isolation using a new enzyme blend, liberase. AB - Enzymatic digestion of donor pancreases is a vital step in human and large mammalian islet isolation. The variable enzymatic activities of different batches of commercially available collagenase is a major obstacle in achieving reproducibility in islet isolation procedures. In the present work, the effectiveness of Liberase, a standardized mixture of highly purified enzymes recently developed for the separation of human islets, was compared with that of a traditional collagenase preparation (type P). The results of 50 islet isolations using Liberase enzyme were compared with those of 36 isolations with collagenase, type P. No significant differences in donor age, cold ischemia time, digestion time, or weight of the pancreases were observed between the two groups. Islet yield was significantly higher in the group where the Liberase enzyme was used. All parameters examined (islet number, islet number per gram of tissue, islet equivalent number, and islet equivalent number per gram of tissue) were significantly improved when Liberase enzyme was used. Different lots of Liberase enzyme were tested, and no difference was observed. Islets isolated with Liberase enzyme were also of larger size and were much less fragmented, suggesting a gentler enzymatic action and better preservation of anatomical integrity. Islets isolated with Liberase enzyme, assessed both in vitro and in vivo, revealed a functional profile similar to that of islets separated with collagenase. Liberase enzyme appears, therefore, to represent a new powerful tool for improving the quality of human islet isolation. PMID- 9200646 TI - Inhibition of diabetes by an insulin-reactive CD4 T-cell clone in the nonobese diabetic mouse. AB - A cloned Th1 cell line was isolated from pancreatic lymph nodes of NOD mice that carries a T-cell receptor encoding Vbeta14 and proliferates in response to NOD islets, islet supernatant, and crystalline bovine and rat insulin, specifically to a B-chain peptide bound to IA(g7). The response to islet supernatant was reduced by 75% by anti-insulin antibody treatment. The insulin-reactive clone reduced insulitis and totally blocked the development of spontaneous diabetes in NOD mice (n = 8) as well as the adoptive transfer of diabetes into irradiated NOD mice following the injection of splenocytes from diabetic mice (n = 13). Trafficking of the adoptively transferred cells was assessed by labeling the clone or diabetic splenocytes with a fluorescent marker (DiI). The labeled clone was detected in the islet periphery, whereas labeled splenocytes alone invaded the islets by 3 days. In contrast, the protective clone dramatically delayed and reduced the number of labeled diabetic splenocytes infiltrating the islet, although their appearance in the spleen was unaffected. In vitro, the clone as well as supernatant derived from the clone blocked the proliferation of diabetic NOD splenocytes to islets. This inhibitory effect was diminished by anti transforming growth factor-beta. In conclusion, an insulin-specific Th1 cell was isolated from NOD mice that traffics to the islet and prevents the spontaneous development and the adoptive transfer of diabetes. It appears to act locally by releasing transforming growth factor-beta and/or other factors that inhibit homing to and/or proliferation of diabetic splenocytes within the islet. These findings may provide insights into and suggest mechanisms for the protective effects of insulin therapy against diabetes. PMID- 9200647 TI - Dietary cow's milk protein does not alter the frequency of diabetes in the BB rat. AB - One theory of the pathogenesis of IDDM proposes that exposure to cow's milk proteins triggers the disease in genetically susceptible individuals. We tested this hypothesis in the BB/Wor rat model of human IDDM. Diabetes-prone (DP) BB/Wor rats spontaneously develop IDDM. Coisogenic diabetes-resistant (DR) BB/Wor rats do not develop diabetes spontaneously, but IDDM can readily be induced by treatment with polyinosinic:polycytidylic acid and depletion of RT6+ T-cells. Pregnant BB/Wor rats were fed one of four experimental diets or a standard Purina commercial rat chow (5010) that was certified to be free of cow's milk protein. Offspring were maintained on the maternal diet after weaning. DP-BB/Wor rats, fed either of two experimental diets based on hydrolyzed casein and free of intact milk protein (Nutramigen or D11236), developed diabetes at only half the rate of animals fed Purina 5010 chow. Neither the addition of bovine serum albumin (BSA) to Nutramigen nor the substitution of total milk protein for the hydrolyzed casein in the D11236 diet increased the frequency of spontaneous diabetes. In contrast, there was no relationship between diet and susceptibility of DR-BB/Wor rats to IDDM induction. However, the methods used to induce IDDM in DR-BB/Wor animals were found to induce antibodies against BSA. We conclude the following: 1) Dietary modification can reduce spontaneous IDDM expression in DP-BB/Wor rats, but the agent of protection is not elimination of cow's milk protein. 2) The addition of BSA or intact milk protein does not abrogate the effectiveness of a protective diet. 3) The genetic susceptibility of the DR-BB/Wor rat to autoimmune diabetes is unaffected by any of the tested diets, but a role of anti-BSA-like autoreactivity in IDDM expression cannot be excluded. PMID- 9200649 TI - Hexokinase shift to mitochondria is associated with an increased sensitivity to glucose in rat pancreatic islets. AB - When rat pancreatic islets are incubated in 5.5 or 16.7 mmol/l glucose for 3 h, an increased sensitivity is observed in islets pre-exposed to high glucose, as indicated by a shift to the left of the glucose dose-response curve (EC50 7.1 +/- 0.9 and 11.5 +/- 1.2 in high- and low-glucose-exposed islets, respectively; n = 5, P < 0.05). To investigate the mechanism(s) responsible for this effect, we measured hexokinase and glucokinase activity both in the cytosolic fraction and in a mitochondrion-enriched fraction, since binding to the outer mitochondrial membrane has been reported to result in an increased enzyme activity. In islets cultured at 16.7 mmol/l glucose, the cytosolic hexokinase activity was similar to control islets, but mitochondrial enzyme activity was significantly increased (124 +/- 7 vs. 51 +/- 9 nmol x microg(-1) x 90 min(-1), P < 0.01). As a consequence, the cytosolic:mitochondrial fraction ratio was altered in comparison with control islets. In contrast, glucokinase activity in the two groups of islets was similar in the cytosolic fraction and undetectable in the mitochondrial fraction. Hexokinase I quantitation by Western blot confirmed the enzyme translocation from the free cytosolic to the mitochondria-bound form in islets cultured at 16.7 mmol/l glucose. Glucose-induced alterations were reversible after 1 h exposure to 5.5 mmol/l glucose. Moreover, in islets exposed to 16.7 mmol/l glucose, inhibition of hexokinase binding to mitochondria by the addition of 20 nmol/l dicyclohexylcarbodiimide resulted in no increase of glucose sensitivity (EC50 10.9 +/- 0.4, n = 3, similar to that of control islets). These data indicate that after chronic exposure to high glucose, the beta-cell becomes more sensitive to glucose before eventually getting desensitized. This increased sensitivity is associated with (and may be due to) an increased hexokinase activity secondary to a subcellular shift of the enzyme from the free cytosolic to the mitochondria-bound, more active form. PMID- 9200648 TI - Glucose stimulates islet beta-cell mitogenesis through GTP-binding proteins and by protein kinase C-dependent mechanisms. AB - Glucose is a cardinal secretory and mitogenic stimulus for the insulin-producing pancreatic beta-cell both in vitro and in vivo, but the mechanisms by which the sugar acts mitogenically remain largely elusive. In this study, the intracellular pathways that convey glucose-induced mitogenic and secretory signaling in beta cells were investigated. For this purpose, fetal rat pancreatic islets enriched in beta-cells were cultured in 3.3 or 16.7 mmol/l glucose for 3 days. It was found that glucose stimulated beta-cell replication, insulin secretion, and cAMP content. These effects were mimicked by agonists of cAMP-dependent protein kinases but not by guanosine-3',5'-cyclic monophosphate (cGMP). Antagonists of cAMP-dependent protein kinases failed to block the glucose-induced increments in beta-cell replication and insulin secretion. Glucose is known to activate protein kinase C, and a protein kinase C-activating phorbol ester was found to promote beta-cell mitogenesis and insulin secretion. Conversely, when protein kinase C was inhibited, the mitogenic (but not secretory) response to glucose was attenuated. There were no additive or synergistic effects on beta-cell replication when cAMP and phorbol ester were combined, whereas insulin secretion was potentiated by this combination. Artificially causing Ca2+ inlet by glibenclamide or ionomycin did not result in a stimulated mitogenic response, and preventing Ca2+ influx by blocking plasma membrane Ca2+ channels did not abolish the mitogenicity of glucose, although it reduced insulin secretion. Pretreatment of islets with pertussis toxin, known to regulate transduction of signals through heterotrimeric GTP-binding proteins, completely prevented the stimulatory effect of glucose on beta-cell mitogenesis but not on insulin secretion. We conclude that specific activation of protein kinase C or cAMP synthesis is sufficient to increase beta-cell mitogenesis and insulin secretion, whereas cGMP appears not to affect these processes. However, cAMP does not seem to mediate the mitogenicity or secretory action of glucose. The results instead suggest that signaling through GTP-binding proteins and protein kinase C activation is required for transduction of the mitogenic, but not secretory, message of the sugar in the beta-cell. PMID- 9200650 TI - Stimulation of islet protein kinase C translocation by palmitate requires metabolism of the fatty acid. AB - The secretory, metabolic, and signaling aspects of glucose/palmitate interaction on beta-cell function have been studied on rat islets. Palmitate potentiated the glucose-induced insulin response of perifused islets at suprathreshold (>3 mmol/l) sugar concentrations. This potentiating effect could be suppressed by 8 bromo-cGMP, which also blocks palmitate metabolism. Palmitate did not modify glucose utilization, but it slightly reduced glucose oxidation and concomitantly increased lactate production. The very low rate of palmitate oxidation (80-fold lower than that of 20 mmol/l glucose) might explain its lack of effect on glycolysis and hence that the glucose/fatty acid cycle is inoperative in islet cells. However, glucose determines the metabolic fate of exogenous palmitate, which is mainly diverted toward lipid synthesis at high sugar concentrations and might then generate lipid messengers for cell signaling. Palmitate did not increase glucose-induced production of inositol-1,4,5-trisphosphate, but it stimulated the translocation of protein kinase C activity from a cytosolic to a particulate fraction at 20 but not at 3 mmol/l glucose. This increased translocation was partially or completely blocked by hydroxycitrate or 8-bromo cGMP, respectively, which are agents interfering with palmitate metabolism (inhibiting lipid synthesis). The metabolic interaction between glucose and palmitate might generate lipid messengers (diacylglycerol, phosphatidylserine) necessary for the activation of islet protein kinase C, which would in turn result in a potentiation of glucose-induced insulin secretion. PMID- 9200651 TI - Voluntary wheel running decreases adipose tissue mass and expression of leptin mRNA in Osborne-Mendel rats. AB - The purpose of this study was to assess the effects of voluntary wheel running on the expression of leptin mRNA in rats that are either sensitive (OM) or resistant (S5B/Pl) to diet-induced obesity. Male OM and S5B/Pl rats had ad libitum access to standard rodent diet and water. At 3-5 weeks of age, animals of both strains were randomly assigned to either an exercise or sedentary control group. The exercise groups had 24-h access to a running wheel, and they trained for 7 weeks. During weeks 1-4, animals in both OM and S5B/Pl exercise groups progressively increased their running. During weeks 5-7, S5B/Pl exercisers tended to run more than did OM (approximately 60 vs. 45 km/week), but by the end of the study both groups had an equally greater heart weight (mg/g body weight) and planteris citrate synthase activity than their sedentary controls. Oral glucose tolerance tests performed during the last week of training revealed that compared with their appropriate controls, insulin sensitivity was enhanced (P < 0.05) in OM but not in the S5B/Pl wheel-running groups. Inguinal, epididymal, and retroperitoneal fat pads weighed less in the running than in the nonrunning groups of both strains (P < 0.01). Additionally, exercised animals had an increased percentage of smaller cells (40-60 microm; P < 0.05) and a decreased percentage of larger cells (120-160 microm; P < 0.05) in the epididymal fat depot. Epididymal leptin mRNA measured by Northern blot analysis was reduced in the exercise-trained rats of both strains (P < 0.05). Furthermore, serum leptin was reduced in exercise trained compared with the control animals of both strains. In comparison to S5B/Pl, control OM animals exhibited both a higher expression and higher circulating levels of leptin (P < 0.05). While serum leptin levels were decreased and food intake was increased in the exercise-trained animals of both strains (P < 0.05), the exact relationship between exercise, leptin, and food intake in this rat model of dietary obesity remains to be determined. Nonetheless, these results suggest that the expression and secretion of leptin can be influenced by exercise training and that these changes (i.e., reduced expression and secretion of protein) can occur independently of changes in whole-body insulin sensitivity and susceptibility to diet-induced obesity. PMID- 9200652 TI - Differences between the tolbutamide-boosted and the insulin-modified minimal model protocols. AB - The insulin-modified frequently sampled intravenous glucose tolerance test (FSIGTT) with minimal model analysis (MINMOD) was compared with the tolbutamide protocol and the glucose clamp in 35 nondiabetic subjects (age 38 +/- 2 years [mean +/- SE], BMI 27.2 +/- 0.9 kg/m2). Each subject underwent two FSIGTTs, one with tolbutamide (300 mg) and the other with insulin (0.03 U/kg) and a euglycemic hyperinsulinemic clamp (40 mU x m(-2) x min(-1)). Insulin sensitivity was determined from each FSIGTT with MINMOD and from the clamp. Insulin sensitivity indexes (S(I)) from the two FSIGTTs were significantly correlated (r = 0.77, P < 0.001), but S(I(insulin)) was 29 +/- 4% lower than S(I(tolbutamide)). Both S(I(insulin)) and S(I(tolbutamide)) correlated significantly with S(I(clamp)) (r = 0.70 and 0.71, P < 0.001 for each). Expressed in the same units (dl/min per pU/ml), S(I(tolbutamide)) was on average 13 +/- 6% lower than S(I(clamp)) (4.51 +/- 0.40 vs. 5.36 +/- 0.36 x 10(-2), P = 0.009), whereas S(I(insulin)) was 44 +/- 4% lower. S(G(tolbutamide)) and S(G(insulin)) were not different (1.88 +/- 0.10 vs. 2.01 +/- 0.09 x 10(-2) min(-1), P = 0.167) and were significantly correlated (r = 0.50, P = 0.002). Thus, insulin sensitivity estimates from both protocols correlate significantly with each other and with the clamp. They are quantitatively discrepant, however, possibly due to differences in the route of insulin delivery, saturation of insulin action, and/or tolbutamide-induced proinsulin release. Data obtained from these two MINMOD protocols are not directly comparable, and the same protocol must be used in any single cross sectional or longitudinal study. PMID- 9200653 TI - Long-term intensive therapy of IDDM patients with clinically overt autonomic neuropathy: effects on hypoglycemia awareness and counterregulation. AB - To test the hypothesis that hypoglycemia unawareness and impaired counterregulation are reversible after meticulous prevention of hypoglycemia in IDDM patients with diabetic autonomic neuropathy (DAN), 21 patients (8 without DAN [DAN-]; 13 with DAN [DAN+]; of the latter, 7 had orthostatic hypotension [DAN+PH+] and 6 did not [DAN+PH-]) and 15 nondiabetic subjects were studied during stepped hypoglycemia (plateau plasma glucose decrements from 5.0 to 2.2 mmol/l) before and 6 months after prevention of hypoglycemia (intensive therapy). After 6 months, frequency of mild hypoglycemia decreased from approximately 20 to approximately 2 episodes/patient-month while HbA1c increased from 6.2 +/- 0.3 to 6.9 +/- 0.2% (P < 0.05). Responses of adrenaline improved more in DAN- patients (from 1.17 +/- 0.12 to 2.4 +/- 0.22 nmol/l) than in DAN+PH- (from 0.75 +/- 0.25 to 1.56 +/- 0.23 nmol/l) and DAN+PH+ patients (from 0.80 +/- 0.24 to 1.15 +/- 0.27 nmol/l, P < 0.05) but remained lower than in nondiabetic subjects (4.9 +/- 0.37 nmol/l, P < 0.05), whereas glycemic thresholds normalized only in DAN-, not DAN+. Autonomic symptoms of hypoglycemia improved but remained lower in DAN- (6.2 +/- 0.6) than in nondiabetic subjects (8.1 +/- 1.1) and lower in DAN+PH+ (4 +/- 0.8) than in DAN+PH- subjects (5.1 +/- 0.8, P < 0.05), whereas neuroglycopenic symptoms normalized (NS). Cognitive function deteriorated less before than after prevention of hypoglycemia (P < 0.05). Thus, intensive therapy with emphasis on preventing hypoglycemia reverses hypoglycemia unawareness in DAN+ patients despite marginal improvement of adrenaline responses, results in low frequency of hypoglycemia despite impaired counterregulation, and maintains HbA1c in the range of intensive therapy. We conclude that DAN, long IDDM duration per se, and antecedent recent hypoglycemia contribute to different extents to impaired adrenaline responses and hypoglycemia unawareness. PMID- 9200654 TI - Long-term effect of lisinopril and atenolol on kidney function in hypertensive NIDDM subjects with diabetic nephropathy. AB - The aim of our study was to evaluate whether inhibition of ACE (lisinopril 10-20 mg/day) can reduce the rate of decline in kidney function more than reducing blood pressure with conventional antihypertensive treatment (atenolol 50-100 mg/day), usually in combination with a diuretic. We performed a prospective, randomized, parallel study for 42 months, double blind for the first 12 months and single blind thereafter. Forty-three (21 lisinopril and 22 atenolol) hypertensive NIDDM patients with diabetic nephropathy were enrolled. Data from 36 patients (17 lisinopril and 19 atenolol, 60 +/- 7 years of age, 27 men) who completed at least 12 months of the study period are presented. At baseline, the two groups were comparable: glomerular filtration rate (51Cr-EDTA plasma clearance) was 75 +/- 6 and 74 +/- 8 ml x min(-1) x 1.73 m(-2), mean 24-h ambulatory blood pressure (A&D TM2420) was 110 +/- 3 and 114 +/- 2 mmHg, and 24-h urinary albumin excretion rate was 961 (range 331-5,727) and 1,578 (476-5,806) mg/24 h in the lisinopril and atenolol groups, respectively. The mean follow-up time was similar, 37 and 35 months in the lisinopril and atenolol groups, respectively. Mean ambulatory blood pressure was equally reduced in the two groups, 12 +/- 2 and 10 +/- 2 mmHg in the lisinopril and atenolol groups, respectively. Glomerular filtration rate declined in a biphasic manner with a faster initial (0 to 6 months) change of 1.25 +/- 0.49 and 0.81 +/- 0.29 ml x min(-1) x month(-1) followed by a slower sustained decline (6 to 42 months) of 0.59 +/- 0.10 and 0.54 +/- 0.13 ml x min(-1) x month(-1) in the lisinopril and atenolol groups, respectively. No significant differences were observed in either initial or sustained decline in glomerular filtration rate between the two groups. Urinary albumin excretion was reduced (% reduction of baseline) more in the lisinopril than in the atenolol group, at 55 (95% CI 29-72) and 15% (-13 to 34), respectively (P = 0.01). In conclusion, the relentless decline in kidney function characteristically found in hypertensive NIDDM patients with diabetic nephropathy can be reduced equally effectively by two antihypertensive treatments, the beta-blocker atenolol and the ACE inhibitor lisinopril. PMID- 9200655 TI - Neural tube defects in embryos of diabetic mice: role of the Pax-3 gene and apoptosis. AB - Neural tube defects are among the most common of the malformations associated with diabetic embryopathy. To study the molecular mechanisms by which neural tube defects occur during diabetic pregnancy, we have developed a new experimental system using pregnant diabetic mice. In this system, the rate of neural tube defects is about three times higher in embryos of diabetic mice than in embryos of nondiabetic mice. Most of the defects affected presumptive midbrain and hindbrain structures and included open defects (i.e., exencephaly) and gross maldevelopment. By semiquantitative reverse transcription-polymerase chain reaction and in situ hybridization, we found that expression of Pax-3, a gene required for neural tube closure in the area of the midbrain and hindbrain, is significantly reduced in the embryos of diabetic mice. The same regions of the neural tube where Pax-3 had been underexpressed were found subsequently to contain high concentrations of cells undergoing apoptosis. Reduced expression of Pax-3 appears to be responsible for this apoptosis because apoptotic cells were also found at sites of neural tube defects in embryos carrying null mutation of the Pax-3 gene. Finally, mouse strains that carry null mutations in Pax-3 develop neural tube defects that resemble the malformations that occur in embryos of diabetic mice. These results suggest that Pax-3 is an important developmental control gene, expression of which is disturbed in embryos of diabetic mice, and that as a consequence, apoptosis of the neural tube occurs. This pathway may be responsible for many of the neural tube defects resulting from diabetic pregnancy. PMID- 9200656 TI - Early, but not advanced, glomerulopathy is reversed by pancreatic islet transplants in experimental diabetic rats: correlation with glomerular extracellular matrix mRNA levels. AB - In this study, we investigated 1) whether long-term restoration of euglycemia by means of pancreatic islet transplants is capable of preventing and/or reversing renal functional and structural alterations in an experimental model of insulin deficient diabetes, and 2) whether changes in extracellular matrix (ECM) and cell turnover at the glomerular level and biochemical abnormalities associated with hyperglycemia correlate with the renal outcome after transplantation. Male Lewis rats, rendered diabetic by intravenous injection of streptozotocin, underwent homologous islet transplantation via the portal vein at 2 weeks (study A), at 4 months (study B), and at 8 months (study C) after the induction of diabetes and killed 12 months after transplantation in study A and 4 months after transplantation in studies B and C. Age-matched nondiabetic and untreated diabetic rats were used as control animals and were studied at 4, 8, and 12 months. In the untreated diabetic animals, metabolic derangement was associated with increased erythrocyte polyol and fructose levels, tail-tendon content of advanced glycation end products (AGEs), total proteinuria, albuminuria, kidney weight, and mean glomerular volume as well as with marked glomerular and extraglomerular lesions. Glomerular gene expression for the ECM components fibronectin and collagen IV and for TGF-beta was also increased, whereas glomerular cell proliferation was unaffected by diabetes. In study A, changes in renal function and structure observed in diabetic rats at 12 months were completely prevented by successful islet transplants. In study B, all functional and structural abnormalities detected in diabetic rats at 4 months of disease duration were virtually reversed by 4 months of euglycemia in transplanted animals, whereas they progressed further in untreated diabetic rats. In study C, the course of functional and structural changes observed in untreated diabetic rats was not reversed by islet transplantation. Likewise, tissue AGE accumulation and particularly upregulation of glomerular ECM and transforming growth factor (TGF)-beta gene expression, which are believed to play a role in the pathogenesis of altered renal function and structure in diabetes, were normalized in transplanted rats from study A and study B, but not in those from study C. These experiments show that restoration of euglycemia by islet transplants is capable of preventing experimental diabetic glomerulopathy and reversing early changes in renal function and structure induced by diabetes. In a later phase of the disease, when glomerular matrix gene expression becomes independent of hyperglycemia, possibly because of the persistent increase in tissue AGE accumulation, metabolic control is not capable of reversing renal abnormalities. PMID- 9200657 TI - Optimization of glycemic control by insulin therapy decreases the proportion of small dense LDL particles in diabetic patients. AB - Small dense LDL particles (B phenotype) are considered to be more atherogenic than large buoyant LDL particles. The influence of glycemic control on LDL particle size and density is still under debate. The aim of this study was to determine LDL subfraction phenotype in both IDDM and NIDDM patients in poor glycemic control compared with that of respective matched control groups. In addition, we evaluated the effect of a 3-month period of optimized glycemic control on this parameter. Thirty-seven IDDM patients and 33 NIDDM patients, together with two respective age-, sex-, and BMI-matched control groups were studied. Non-A phenotype prevalence in IDDM patients before (19%) and after blood glucose optimization (11%) was similar to that of their control group (12%). However, NIDDM patients displayed a higher proportion of the non-A phenotype (51%) than did the control group (28%), but it became closer (30%, P < 0.05) after glycemic control improved. All subjects with non-A phenotype that changed to A phenotype showed triglyceride levels below 1.63 mmol/l and a greater decrease in HbA1c than did subjects whose phenotype did not change (4.9 +/- 1.5 vs. 3.1 +/- 1.4%, P < 0.05). A higher proportion of small dense LDL was observed in NIDDM women than in nondiabetic women (LDL5 10.0 +/- 4.8 vs. 6.3 +/- 1.5%, LDL6 6.1 +/- 2.2 vs. 4.2 +/- 0.8%, P < 0.05) during both stages of glycemic control, but no differences were observed between NIDDM and nondiabetic men. In conclusion, these findings provide new evidence for the relevance of near-normal glycemic control in the prevention of macrovascular disease and could contribute to an explanation of the loss of protection for cardiovascular disease in diabetic women. PMID- 9200659 TI - Mapping NIDDM susceptibility loci in French families: studies with markers in the region of NIDDM1 on chromosome 2q. PMID- 9200658 TI - Increased synthesis of tumor necrosis factor-alpha in uterine explants from pregnant diabetic rats and in primary cultures of uterine cells in high glucose. AB - The production of tumor necrosis factor-alpha (TNF-alpha) was investigated in uterine explants from normal, diabetic, or insulin-treated diabetic pregnant rats. Explants from diabetic rats released more soluble TNF-alpha than did those in the other groups. The extent of this secretion was correlated with blood glucose concentration at the time of explantation. The concentration of cell membrane-associated TNF-alpha in the explants was not altered by diabetes. Daily insulin administration failed to normalize uterine TNF-alpha secretion despite correction of glycemia in the diabetic rats. Explants from normal pregnant rats cultured in vitro with increasing concentrations of D-glucose showed a dose dependent increase in TNF-alpha secretion. The production of TNF-alpha in high glucose was also tested in primary cultures of uterine cells isolated from either immature or adult rats. TNF-alpha secretion was increased in high D-glucose but not in iso-osmolar concentrations of L-glucose, D-raffinose, D-galactose, or mannitol. Cell membrane-associated TNF-alpha was not influenced by high D glucose. Semiquantitative reverse transcription-amplification of RNA extracted from primary cultures of uterine cells showed that the steady-state level of TNF alpha transcripts was increased by high D-glucose but not by high L-glucose. The results are consistent with the hypothesis that hyperglycemia is instrumental in the overexpression of TNF-alpha in the diabetic uterus. Because TNF-alpha has a demonstrated negative impact on embryonic growth, enhanced TNF-alpha synthesis in the pregnant uterus may contribute to the embryopathy associated with maternal diabetes. PMID- 9200660 TI - The 31-cM region of chromosome 11 including the obesity gene tubby and ATP sensitive potassium channel genes, SUR1 and Kir6.2, does not contain a major susceptibility locus for NIDDM in 127 non-Hispanic white affected sibships. PMID- 9200661 TI - PPAR-gamma gene expression is elevated in skeletal muscle of obese and type II diabetic subjects. AB - The peroxisome proliferator activated receptor PPAR-gamma has been identified as a nuclear receptor for thiazolidenediones, which are compounds with insulin sensitizing properties in several tissues, including skeletal muscle. To determine whether this receptor is expressed and possibly involved in insulin action/resistance in skeletal muscle, PPAR-gamma mRNA abundance and its regulation by insulin were quantified in muscle tissue and cultures from lean and obese nondiabetic and type II diabetic subjects using competitive reverse transcription-polymerase chain reaction (RT-PCR). In muscle biopsy specimens, PPAR-gamma mRNA was elevated in obese nondiabetic and type II diabetic subjects (23.4 +/- 4.2 and 28.0 +/- 5.69 x 10(3) copies/microg total RNA, respectively; both P < 0.05) compared with lean nondiabetic control subjects (9.4 +/- 2.3 x 10(3) copies/microg total RNA). Significant positive correlations were present among skeletal muscle PPAR-gamma mRNA levels, BMI (r = 0.67, P < 0.01), and fasting insulin concentration (r = 0.76, P < 0.001). PPAR-gamma mRNA levels were also elevated in muscle cultures from type II diabetic subjects compared with lean nondiabetic control subjects (330.1 +/- 52.9 vs. 192.1 +/- 27.0 x 10(3) copies/microg total RNA, P < 0.05). Insulin stimulation of muscle tissue (by hyperinsulinemic-euglycemic clamp for 3-4 h) or muscle cultures (30 nmol/l for 120 min) stimulated PPAR-gamma mRNA expression up to fourfold (10.0 +/- 2.7 to 41.3 +/- 7.4 x 10(3) copies/microg total RNA, P < 0.05, and 174.9 +/- 56.9 to 268.2 +/- 78.6 x 10(3) copies/microg total RNA, P < 0.05, respectively). In summary, PPAR-gamma mRNA expression in human skeletal muscle is acutely regulated by insulin and is increased in both obese nondiabetic and type II diabetic subjects in direct relation to BMI and fasting insulinemia. We conclude that abnormalities of PPAR-gamma may be involved in skeletal muscle insulin resistance of obesity and type II diabetes. PMID- 9200662 TI - Evidence for a novel peripheral action of leptin as a metabolic signal to the adrenal gland: leptin inhibits cortisol release directly. AB - The crucial role of glucocorticoids in obesity and insulin resistance and the actions of the OB protein leptin on the hypothalamic-pituitary-adrenal (HPA) axis suggest that there is an important interaction of leptin with the glucocorticoid system. Therefore, we designed a study to test the effect of leptin directly on adrenocortical steroidogenesis. Primary cultures of bovine adrenocortical cells were incubated with increasing concentrations (10-1,000 ng/ml) of recombinant mouse leptin for 24 h, and the effects of leptin on basal and ACTH-stimulated cortisol secretion were determined. The accumulation of P450 17alpha mRNA following incubation with ACTH (10 nmol/l) and leptin (10-1,000 ng/ml) was analyzed by Northern blot. Adrenocortical cells were characterized by immunohistochemical staining for 17alpha-hydroxyprogesterone. Leptin (10-1,000 ng/ml) inhibited basal and ACTH-stimulated cortisol release. At a concentration that occurs in obese individuals in vivo (100 ng/ml), it reduced basal cortisol secretion to 52.7 +/- 37% (mean +/- SE). The rise in cortisol secretion following maximal ACTH stimulation (10 nmol/l) was blunted to 55.2 +/- 27%. At more physiological concentrations of ACTH (0.1 nmol/l), the inhibition of cortisol release by coincubation with low doses of leptin (10 ng/ml) was even more pronounced, leading to a reduction to 32.8% (1,248 +/- 134 vs. 410 +/- 157 nmol/l). Addition of OB protein (10-1,000 ng/ml) led to a dose-dependent reduction of ACTH-stimulated cytochrome P450 17alpha mRNA accumulation (from 80 to 45%), suggesting that leptin regulates adrenal steroidogenesis at the transcriptional level. These data clearly demonstrate that leptin inhibits cortisol production in adrenocortical cells and therefore appears to be a metabolic signal that directly acts on the adrenal gland. PMID- 9200663 TI - Melanocortins and opiate addiction. AB - Adrenocorticotropic hormone (ACTH) and alpha-melanocyte stimulating hormone (alpha-MSH) are centrally acting melanocortin peptides with numerous reported functions, including induction of excessive grooming and antipyresis, among others. Also reported is a role for melanocortins in aspects of opiate action. Although early work examined the effects of ACTH and MSH on opiate-induced behaviors, further progress has been limited. Recently, however, advances in the identification and characterization of melanocortin receptor (MC-R) subtypes have provided novel tools with which to study interactions between melanocortins and addiction. The present review discusses the effects of ACTH and MSH on opiate induced behaviors and relates these findings to more recent reports on the regulation of melanocortin systems by exogenous opiates. Emerging from these data is the possibility that melanocortin receptor activation, specifically at the MC4 R subtype, may act to antagonize certain properties of exogenous opiates, including perhaps addiction. PMID- 9200664 TI - Discriminative stimulus properties of eltoprazine. AB - Rats were trained to discriminate eltoprazine (1-(2,3-dihydro-1,4-benzodioxin-5 yl)-piperazine) (1.0 mg/kg p.o.) from demineralized water in a two lever operant procedure. Eltoprazine generalized to the 5-HT1B receptor agonist anpirtoline (6 chloro-2-[piperidyl-4-thiol]-pyridine hydrochloride), the 5-HT(1A,1B) receptor agonists batoprazine (8-(1-piperazinyl)-2H-1-benzopyran-2-one) and 1-NP (1-(1 naphthyl)piperazine hydrochloride), and to the 5-HT(1B/2C) receptor agonist mCPP (1-(3-chlorophenyl)piperazine dihydrochloride). The 5-HT1A receptor agonist flesinoxan (R(+)-N-[2[4-(2,3-dihydro-2-2-hydroxy-methyl-1,4-benzodioxin-5-yl) -1 piperazinyl]ethyl]-4-fluorobenzoamide) generalized partially and the 5-HT1A receptor antagonist WAY-100635 (N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N (2-pyridinyl) cyclohexanecarboxamide trihydrochloride) failed to antagonize the eltoprazine cue, suggesting that 5-HT1A receptors are of limited importance in the discriminative stimulus properties of eltoprazine. Methiothepin, mCPP, mianserin and alprazolam did not antagonize the eltoprazine cue. The 5 HT(1A,1B,1D) receptor agonist GR46611X (3-[3-(2-dimethylamino-ethyl)-1H-indol-6 yl]-N-(4-methoxy-benzyl)acrylam ide) and the 5-HT(1B,1D) receptor antagonist GR127935T (N-[4-methoxy-3-(4-methyl-1-piperazinyl) phenyl]-2'-methyl-4'-(5-methyl 1,2,4-oxadiazol-3-yl) [1,1,-biphenyl]-4-carboxamide) did neither generalize to nor antagonize the eltoprazine cue, whereas (-)-alprenolol showed partial antagonism and substitution. These results show that the eltoprazine discriminative stimulus is mediated by the 5-HT1B receptor, although the lack of good 5-HT1B receptor antagonists weakens this conclusion. PMID- 9200665 TI - Influence of growth hormone on cysteamine-induced gastro-duodenal lesions in rats: the involvement of somatostatin. AB - It is known that cysteamine ulcerogenic effect depends, among others, on a depletion of somatostatin in the gastro-intestinal tract. Since growth hormone (GH) causes the release of hypothalamic somatostatin (SRIH) and potentiates the ulcerogenic action of cysteamine we have studied the influence of GH on gastro duodenal mucosa levels of SRIH, and its relevance for the ulcerogenic action of cysteamine. Female rats of the Sprague-Dawley strain were pretreated with GH (0.25, 0.5 or 1 mg/kg) and subjected to cysteamine-induced gastric lesions. These animals showed an increased mortality and severity of gastric lesions. The measurement of gastric and duodenal barrier mucus levels revealed that GH administration was followed by a decrease in mucus production. Pretreatment with SRIH (25 or 50 microg/kg) was followed by a decreased percent incidence and severity of gastric mucosa lesions induced by cysteamine. The mucus production was increased by SRIH administration. GH pretreatment was followed by a reduction of SRIH-like immunoreactivity in gastro-duodenal mucosa and an increase of insulin plasma levels. Acute injection of cysteamine per se was also followed by a decrease of gastro-duodenal SRIH and an increase in insulin plasma levels. These results suggest that high levels of plasma GH, as induced by exogenous GH administration, may cause a decrease of SRIH gastro-intestinal content and this in turn may potentiate the ulcerogenic activity of cysteamine. PMID- 9200666 TI - Dual action of colchicine on hypertonic activation of system A amino acid transport in vascular smooth muscle cells. AB - Amino acid transport system A is present in many cells and tissues and is regulated by hormones and other factors, including hypertonic stress. System A in vascular smooth muscle cells is also activated when microtubules are disrupted by drugs such as colchicine. The present study examined the action of colchicine on hypertonic activation of system A in smooth muscle cells. In serum-free medium, activation of system A by modest (340 mOsm) hypertonicity was not affected by colchicine addition. However, at high osmotic stress (460 mOsm) the addition of colchicine partially blocked the activation of system A. Addition of colchicine alone, at normal osmolarity, produced activation of system A. In the presence of serum, colchicine action was markedly different. Colchicine consistently inhibited hypertonic activation of system A at any degree of hypertonic stress but had no effect on system A at normal osmolarity. The action of colchicine as both an activator and inhibitor of system A implies microtubule involvement at more than one step in the intracellular regulation of system A. PMID- 9200667 TI - Metabolic alterations in muscle of thermally injured rabbits, measured by positron emission tomography. AB - The hypermetabolic inflammatory state that occurs after major trauma has been extensively studied at the whole body level, however, there is only limited information on metabolic changes in individual tissues. In this study, the effect of thermal injury on metabolic function of uninjured hind limb muscle of rabbits was measured noninvasively by positron emission tomography (PET). Rabbits were subjected to full thickness burn on 25% of their body surface area. Two to three weeks after injury, PET and arterial blood sampling was performed during inhalation of 15O2, C15O2 and 11CO and after injection of 18FDG. The tissue and blood data were analyzed by standard kinetic models for blood flow, oxygen extraction fraction (OEF), oxygen utilization and glucose metabolism. A total of seven injured and five sham animals were studied. Total body oxygen consumption was measured by indirect calorimetry and plasma concentrations of glucose, insulin and IGF-1 were measured with standard assays. Compared to sham rabbits, blood flow to muscle of injured animals was unchanged. However, OEF, oxygen utilization and glucose metabolism were significantly reduced (p<0.01) in uninjured muscle of burned rabbits. These data demonstrate that thermal injury is associated with alterations in muscle metabolism, which are not related to change in blood flow. PMID- 9200668 TI - Inhibition of nitric oxide facilitates LH release from rat pituitaries. AB - We examined the effects of nitric oxide (NO) modulators on rat pituitary LH content in vivo and studied their response to LHRH-stimulated LH secretion in vitro in ovariectomized adult female Sprague Dawley rats. Alzet mini pumps (flow rate 10 microl/h) delivering either normal saline (Group I, 1.2 mg nitroglycerin, a donor of NO (Group II) or 50 mg of nitro-L-Arginine methyl ester, a NO synthase (NOS) inhibitor (Group III), were subcutaneously implanted into experimental animals. Following 36 h infusion, pituitaries were removed and either frozen for LH quantitation, or fragmented and challenged in the superfusion system with 10 min pulses of LHRH (1 ng/ml) at 90 min intervals for 10 hours. LH was assayed by radio-immunoassay (RIA) in the homogenates of pituitaries and in aliquots of the superfusate collected every 10 mins. Significantly lower pituitary LH levels were noted in Group III (150.3 +/- 18.6 ng) in comparison to Groups I (215.6 +/- 5.5 ng; p<0.04) or II (221.2 +/- 14.9 ng; p<0.01), suggesting that low levels of NO stimulate LH secretion in vivo. The pituitary LH contents were not significantly different in Groups I and II. In vitro studies reveal that exogenous LHRH stimulated response, measured as average pulse response (90 minute period after LHRH), and total LH released during the 10 hour perfusion, was 290 +/- 23.6 ng and 1646.7 +/- 270.8 ng, respectively, in Group III; 57.9 +/- 3.1, and 344.7 +/- 24.3 ng in Group I, and 105.3 +/- 6.3, and 633.7 +/- 77.1 mg in Group II. Thus, our in vitro studies demonstrate significantly enhanced (p<0.05) LHRH- stimulated LH secretion in Group III in comparison to Groups I and II, while Group II shows higher responsiveness than Group I (p<0.05). The results of the current studies provide evidence that NOS inhibition facilitates pituitary LH secretion. The differential responses to LHRH-stimulated LH secretion in vitro in the 3 groups suggest a possible role of NO in modulating pituitary LHRH receptor concentrations. However, this will have to be tested by further studies. PMID- 9200669 TI - Oral insulinlike growth factor-I stimulates intestinal enzyme maturation in newborn rats. AB - Insulinlike growth factor-I (IGF-I) has been found in the milk of various species. To investigate if milk-borne IGF-I has any effect on postnatal gut development in neonatal animals, newborn rat pups were given orally 1 microg recombinant human IGF-I daily for 3 days. For comparison, a separate group of newborn pups was given 150 microg hydrocortisone, the hormone known to stimulate intestinal maturation in neonatal rats. Oral IGF-I treatment had no significant effect on the animal body weight nor on the weight of the stomach, small and large intestines, and pancreas. Oral administration of hydrocortisone significantly reduced body weight gain, but it had no apparent effect on internal organ weights. Both IGF-I and hydrocortisone treatments, however, significantly increased lactase, maltase and sucrase activities and hydrocortisone significantly increased aminopeptidase activity at the proximal small intestine when compared with the control. The finding supports the hypothesis that milk borne IGF-I may play a role in regulating postnatal gut development in the suckling young. PMID- 9200670 TI - Beta-3 adrenoceptor (beta-3AR) expression in leptin treated OB/OB mice. AB - Leptin-deficient Ob/Ob mice are hypometabolic and have reduced fat cell expression of beta-3 adrenoceptors (ARs). To determine whether leptin repletion restores beta-3 AR number, C57BL/6J Ob/Ob mice were given exogenous leptin (5 mg/kg I.P. daily) for 21 days. Leptin administration reduced body weight from 43.1+/-3.7 to 34.1+/-3.7 g in Ob/Ob animals but had no effect on weight in wildtype animals. Body weight increased by 12% in Ob/Ob mice receiving saline. Beta-3 AR mRNA concentrations were markedly reduced in Ob/Ob animals at baseline. Leptin increased beta-3 AR mRNA to control levels in Ob/Ob mice, but had no effect in wildtype animals. Adipocyte leptin mRNA was increased by 400% in Ob/Ob mice and did not suppress with exogenous leptin administration, suggesting no direct feedback regulation of leptin synthesis. We speculate that restoration of beta-3 AR expression by repleting leptin may be important in correcting hypometabolism in Ob/Ob animals. PMID- 9200671 TI - The role of catecholamines in the etiology of infertility in diabetes mellitus. AB - Patients suffering from diabetes mellitus often develop reproductive dysfunction including anovulation, infertility and disrupted pregnancy. The biochemical basis of these phenomena is yet to be provided. The current study utilizes a neuroendocrine paradigm involving an in vitro microdissection technique in conjunction with jugular catheterization to examined the proestrus dynamics of norepinephrine (NE) and the preovulatory luteinizing hormone (LH) surge in streptozotocin (STZ) treated female rats, an animal model for insulin dependent diabetes mellitus. Radioimmunoassays revealed that in control subjects LH was at basal level during the morning of proestrus (900-1200 h); the first significant increase in the level of this pituitary hormone occurred at 1400-1500 h. A maximum peak concentration of LH was attained at 1700 h. In contrast, plasma levels of LH in diabetic subjects showed the first significant increase at 1500 h and peaked at 2000 h. The peak of the LH curve in diabetic rats was reduced by about 65% with a 3 h shift to the right. Alpha-methyl-p-tyrosine-induced blockade of newly synthesized NE-based assay showed that NE turnover rates in several hypothalamic nuclei (e.g. medial preoptic nucleus, MPN; median eminence, ME; suprachiasmatic nucleus, SCN; arcuate nucleus, AN) of control subjects were at basal level during the morning of proestrus (0900-1100 h). However, they increased by the 1200-1400 h interval and remained elevated during the 1500-1700 h. This time dependent increase in hypothalamic NE turnover rates during proestrus was not observed in the STZ diabetic rats. Most of the above metabolic derangements were partially reversed following the institution of insulin replacement therapy. Overall, our data support the concept that the endocrine abnormalities (e.g. infertility, delayed preovulatory LH surge) in diabetes are due, at least in part, to a functional deficit in noradrenergic neurons within the hypothalamus. PMID- 9200672 TI - Monometoxytrityl derivatives of uridine as inhibitors of a human recombinant UDP glucuronosyltransferase: UGT1*6. AB - A series of inhibitors of the human liver recombinant UDP-glucuronosyltransferase 1*6 derived from uridine were synthetized as probes of the binding site of the cosubstrate, UDP-glucuronic acid. If triphenylmethanol or uridine alone failed to inhibit the glucuronidation of 4-methylumbelliferone, the trityl derivatives of uridine were found to be very effective inhibitors of the enzyme (Ki 4.4 to 73 microM). The type of inhibition (competitive or mixed) varied with the substitutions on the uracile or on the triphenylmethyl moiety by halogen atoms or methyl groups. Structural features for the binding of the cofactor are postulated. PMID- 9200673 TI - Evaluation of a Cys23Ser mutation within the human 5-HT2C receptor gene: no evidence for an association of the mutant allele with obesity or underweight in children, adolescents and young adults. AB - Serotonin is a neurotransmitter involved in a large number of psychophysiological processes including the regulation of mood, arousal, aggression, sleep, learning, nociceptions, nerve growth and importantly, appetitive functions. Alterations of 5-HT receptor activity have been shown to occur in many psychiatric diseases including depression, anxiety, eating disorders, schizophrenia etc. Hence, genetic variation in genes coding for serotonin receptor proteins might well be involved in the genetic predisposition to these diseases and therefore are of great pharmacogenetic relevance. Knockout mice deficient of a functional 5-HT2C receptor have implicated a potential role of this receptor subtype in the serotonergic control of appetite. A Cys23Ser mutation in the human 5-HT2C receptor gene discovered recently prompted us to investigate this mutation with regard to the development of human obesity. We have evaluated this mutation in 241 obese children and adolescents (mean BMI > or = 97th percentile), 80 normal weight children (BMI 5th-85th percentile) and 92 underweight probands (BMI < or = 15th percentile) for a possible association with obesity. The frequencies of the mutant allele in all three weight groups (obese subjects: 0.1597; normal weight: 0.168; underweight: 0.1575) were very similar. Association as well as linkage studies were negative. Therefore it is unlikely that this receptor mutation plays a direct role in the development of human obesity. PMID- 9200674 TI - AT1 receptors mediate pressor responses induced by angiotensin II in the periaqueductal gray area of rats. AB - Microinjection of angiotensin II (ANGII) (0.01 to 1 nmol) into the periaqueductal gray area (PAG) of anaesthetised rats caused dose-dependent increases in blood pressure. Preinjection (10 min before) of losartan (a selective non-peptide AT1 receptor antagonist; 50 nmol) to the PAG reduced the pressor response to ANGII whereas PD123319 (a selective non-peptide AT2 receptor antagonist; 50 nmol) did not affect the ANGII-induced hypertension. Thus, our data suggest that the activation of AT1 but not AT2 receptors mediates ANGII-induced blood pressure changes in the PAG area. PMID- 9200675 TI - The effect of ovariectomy on the contractile response of the rat isolated detrusor muscle and urethra. AB - Contractile responses induced by carbachol on the detrusor muscle and by noradrenaline on the isolated urethra were compared between ovariectomized rats pretreated with estradiol (50 microg/animal s.c. twice daily for five days), untreated ovariectomized rats and intact animals. In the detrusor muscle, contractions induced by 30 microM carbachol, when normalized with respect to KCl 100 mM-induced contraction, were similar for the three groups. Furthermore, contractions induced by 100 microM noradrenaline in the isolated urethra were not significatively different between groups. However, the pD2 value for noradrenaline was greater in urethral tissue from ovariectomized rats compared with ovariectomized -estrogen treated and control rats. A similar result was found for pD2 values for carbachol-induced contractions on the detrusor muscle. These results suggest that ovariectomy increases the sensitivity of the urinary bladder and urethra to the contractile effects of carbachol and noradrenaline, respectively and that this effect is reversed by in vivo estrogen pretreatment. PMID- 9200676 TI - Chemical versatility of transplatin monofunctional adducts within multiple site specifically platinated DNA. AB - The first step of the reaction between DNA and the antitumor drug cisplatin or its clinically inactive isomer transplatin yields monofunctional adducts. Most of the cisplatin monofunctional adducts further react and rather rapidly (t(1/2) smaller than a few hours) to form intrastrand and interstrand cross-links. It is generally accepted that the clinical activity of cisplatin is related to the formation of bifunctional lesions. As concerns transplatin, several studies disagree on the rate of closure of the monofunctional adducts and the nature of the bifunctional lesions. In order to explain these discrepancies, we have prepared several duplexes containing a single monofunctional trans [Pt(NH3)2(dG)Cl]+ adduct and zero to two monofunctional [Pt(dien)(dG)]2+ adducts at defined positions. In these duplexes, the inert [Pt(dien)(dG)]2+ adducts mimic the presence of transplatin monofunctional adducts. We show that the closure of the transplatin monofunctional adducts is strongly affected by the presence of other adducts and by the length of the duplexes. These findings suggest that the discrepancies in the literature originate from the nature of the platinated samples (molar ratio of bound platinum per nucleotide, length of the DNA fragments). Our general conclusion is that within transplatin-modified DNA, at a low level of platination, the monofunctional adducts evolve slowly (t(1/2) > 24 h) into bifunctional lesions and that these bifunctional lesions are mainly interstrand cross-links. This could explain, at least in part, the clinical inefficiency of transplatin. PMID- 9200677 TI - 13C Magic angle spinning NMR analysis and quantum chemical modeling of the bathochromic shift of astaxanthin in alpha-crustacyanin, the blue carotenoprotein complex in the carapace of the lobster Homarus gammarus. AB - Selective isotope enrichment, 13C magic angle spinning (MAS) NMR, and semiempirical quantum chemical modeling, have been used to analyze ligand-protein interactions associated with the bathochromic shift of astaxanthin in alpha crustacyanin, the blue carotenoprotein complex from the carapace of the lobster Homarus gammarus. Spectra of alpha-crustacyanin were obtained after reconstitution with astaxanthins labeled with 13C at positions 4,4', 12,12', 13,13', or 20,20'. The data reveal substantial downfield shifts of 4.9 and 7.0 ppm at positions 12 and 12' in the complex, respectively. In contrast, at the 13 and 13' positions, small upfield shifts of 1.9 ppm were observed upon binding to the protein. These data are in line with previously obtained results for positions 14,14' (3.9 and 6.8 ppm downfield) and 15,15' (0.6 ppm upfield) and confirm the unequal perturbation of both halves after binding of the chromophore. However, these results also show that the main perturbation is of symmetrical origin, since the chemical shift differences exhibit a similar pattern in both halves of the astaxanthin molecule. A small downfield shift of 2.4 ppm was detected for the 4 and 4' positions. Finally, the 20,20' methyl groups are shifted 0.4 ppm upfield by the protein. The full data set provides convincing evidence that charge polarization is of importance for the bathochromic shift. The NMR shifts are compared with calculated charge densities for astaxanthin subjected to variations in protonation states of the ring-functional groups, as models of ligand-protein interactions. Taking into account the color shift and other available optical data, the current model for the mechanisms of interaction with the protein was refined. The results point toward a mechanism in which the astaxanthin is charged and subject to strong electrostatic polarizations originating from both keto groups, most likely a double protonation. PMID- 9200679 TI - Anisotropic molecular rotational diffusion in 15N spin relaxation studies of protein mobility. AB - The backbone dynamics of the uniformly 15N-labeled N-terminal 63-residue DNA binding domain of the 434 repressor has been characterized by measurements of the individual 15N longitudinal relaxation times, T1, transverse relaxation times, T2, and heteronuclear 15N[1H]-NOEs at 1H resonance frequencies of 400 and 750 MHz. The dependence of an apparent spherical top correlation time, tauR, on the orientation of the N-H bond vector with respect to the principal axes of the global diffusion tensor of the protein was used to establish the fact that the degree of anisotropy of the global molecular tumbling amounts to 1.2, which is in good agreement with the values obtained from model calculations of the hydrodynamic properties. A model-free analysis showed that even this small anisotropy leads to the implication of artifactual slow internal motions for at least two residues when the assumption of isotropic global motion is used. Additional residues may actually undergo internal motions on the same time scale as the global rotational diffusion, in which case the model-free approach would, however, be inappropriate for quantifying the correlation times and order parameters. Overall, the experiments with 434(1-63) demonstrate that the assumption of isotropic rotational reorientation may result in artifacts of model free interpretations of spin relaxation data even for proteins with small deviations from spherical shape. PMID- 9200678 TI - Protein-protein interaction of the human poly(ADP-ribosyl)transferase depends on the functional state of the enzyme. AB - Poly(ADP-ribosyl)transferase (pADPRT) is a nuclear protein which catalyzes the polymerization of ADP-ribose using NAD+ as substrate, as well as the transfer of ADP-ribose polymers to itself and other protein acceptors. The catalytic activity of pADPRT strictly depends on the presence of DNA single-strand breaks. In this report, protein-protein interaction of pADPRT was found to depend on both the extent of automodification with poly(ADP-ribose) and the presence of DNA. Specific binding of radiolabeled pADPRT to transblotted proteins was first tested in blot overlay experiments. For radiolabeling, use was made of the ability of the enzyme to incorporate [32P]ADP-ribose from [32P]NAD+. Varying the concentration of NAD+, two different forms of automodified pADPRT were obtained: oligo(ADP-ribosyl)ated pADPRT with less than 20 ADP-ribose units per chain, and poly(ADP-ribosyl)ated pADPRT with polymer lengths of up to 200 ADP-ribose residues. Interaction of these probes with transblotted HeLa nuclear extracts, purified histones, and distinct regions of recombinant pADPRT was investigated. While the oligo(ADP-ribosyl)ated enzyme associated preferentially with transblotted purified histones, or pADPRT present in HeLa nuclear extracts, poly(ADP-ribosyl)ated pADPRT bound to a variety of transblotted proteins in the nuclear extracts. In the presence of DNA, both the oligo- and the poly(ADP ribosyl)ated enzymes bound to the transblotted recombinant zinc finger domain of pADPRT even at high salt concentrations. In the absence of DNA, the transblotted automodification domain of pADPRT appeared to be the region involved in self association. In another set of experiments, unmodified or poly(ADP-ribosyl)ated pADPRT was immobilized on Sepharose. Affinity precipitation of recombinant pADPRT domains confirmed the specific interaction of pADPRT with its zinc finger region and the automodification domain, whereas no interaction was observed with the NAD+ binding domain. Affinity precipitation of HeLa nuclear extracts with poly(ADP-ribosyl)ated pADPRT-Sepharose led to the enrichment of a number of proteins, whereas nuclear proteins bound to the unmodified pADPRT-Sepharose in a smaller extent. The results suggest that protein-protein interaction of the human pADPRT is governed by its functional state. PMID- 9200680 TI - Molecular dynamics simulations of the unfolding of barnase in water and 8 M aqueous urea. AB - Molecular dynamics simulations of barnase have been conducted both in water and in 8 M urea solution for 500 ps at 25 degrees C and for 2000 ps at 85 degrees C. The final structure of the aqueous simulation at room temperature matches closely the structure obtained by NMR and the experimentally observed protections from isotopic exchange. The comparison of the structures generated by the aqueous simulation at 85 degrees C reveals a trajectory composed of groups of geometrically related structures separated by narrow regions of rapid change in structure. The first of these regions displays changes in backbone rmsd to the crystal structure and solvent-accessible area suggestive of a transition state, while the properties observed during the final 300 ps of the simulation are consistent with a stable intermediate. These assignments were confirmed by calculation of the "progress along the reaction coordinate" phi-values using an empirical equation based on a linear response method. The pathway of unfolding defined in this fashion agrees well with the experimental results of site directed mutagenesis in terms of secondary structure content of the transition state and the intermediate and reproduces the relative stability of the different elements of secondary structure. The results of the simulations in urea suggest a mechanism at the molecular level for its well-known enhancement of the denaturation of proteins. The analysis of radial distribution functions shows that the first solvation shell of the protein is enriched in urea relative to the bulk solvent. The displacement of water molecules allows greater exposure of hydrophobic side chains, as witnessed particularly in the analysis of solvent accessible surface areas at the higher temperature. Almost all urea molecules in the first shell form at least one hydrogen bond with the protein. They provide a more favorable environment for accommodation of the remaining water molecules, and they facilitate the separation of secondary structure elements by acting as a bridge between groups previously forming intraprotein hydrogen bonds. PMID- 9200681 TI - Three-dimensional reconstruction of native and reassembled Lumbricus terrestris extracellular hemoglobin. Localization of the monomeric globin chains. AB - The approximately 3.5 MDa hexagonal bilayer (HBL) hemoglobin (Hb) of the earthworm Lumbricus terrestris is composed of monomers and disulfide-bonded trimers (T) of globin chains and of four types of heme-deficient linker chains (L). Cryoelectron microscopic images of native Hb and of HBL reassembled from the constituent subunits depleted in monomer subunit (HBL[T+L]) were subjected to three-dimensional reconstructions by the random conical tilt series method. Native Hb has an architecture very similar to those of other annelid and vestimentiferan Hbs, consisting of 12 hollow globular substructures (HGS). Each HGS is comprised of six dense masses, has a 3-fold symmetry, and is organized in two hexagonally symmetric layers, with the vertices of the upper layer rotated 16 degrees clockwise relative to those of the lower layer. The layers are tethered to a central linker complex, consisting of two bracelets of connections perpendicular to the 6-fold axis and a set of six vertical connections linked to a flat hexagonal mass. HBL[T+L] shared all these features with the native Hb, except for a large hole around the 3-fold symmetry axis in each HGS, indicating the probable location of the missing monomer subunit. PMID- 9200682 TI - Immunosuppressor binding to the immunophilin FKBP59 affects the local structural dynamics of a surface beta-strand: time-resolved fluorescence study. AB - The interaction of the immunophilin domain of FKBP59 (FKBP59-I) with immunosuppressant drugs was investigated by steady-state and time-resolved fluorescence of tryptophan. One of the two Trp residues present in this protein (W89), conserved in almost all immunophilins, is buried in the hydrophobic core and participates in the immunosuppressant binding. By comparison with the highly homologous protein FKBP12, containing only the buried Trp, it has been concluded that its weak fluorescence is due to an atypical H-bond interaction involving the indole nitrogen and the Phe129 benzene ring. The second Trp residue (W59) in FKBP59-I is located on the external hydrophilic side of the 50-60 beta-sheet [Craescu, C. T., Rouviere, N., Popescu, A., Cerpolini, E., Lebeau, M.-C., Baulieu, E.-E., & Mispelter, J. (1996) Biochemistry 35, 11045-11052] and is responsible for >95% of the fluorescence emission. The long lifetime of the major excited state, the large activation energy of thermal quenching, and the rotational correlation time distribution pattern suggest that its environment is not highly mobile. Binding of the immunosuppressant drugs FK506 and rapamycin leads to a approximately 60% decrease of the fluorescence intensity without any change in the fluorescence emission maximum. Time-resolved measurements show that this "quenching" is due to a conformational change which depletes the long excited-state lifetime population to the profit of a more quenched minor excited state, which becomes prominent in the complexes. This is accompanied by a strong slowing of the indole ring dynamics in the case of FK506 and by a complete immobilization in the case of rapamycin, as shown by two-dimensional (tau, theta) maximum entropy analysis of the polarized fluorescence decays. Binding of the immunosuppressant drugs therefore modifies the structure and the dynamics of the external side of the 50-60 beta-sheet in FKBP59-I, which could be relevant for the formation of ternary complexes with other protein targets. PMID- 9200683 TI - A conformational change in F-actin when myosin binds: fluorescence resonance energy transfer detects an increase in the radial coordinate of Cys-374. AB - Interactions of myosin with actin filaments probably induce conformational changes in actin which are crucial for its function. Fluorescence resonance energy transfer spectroscopy can determine changes in distance (range 10-100 A) between two probes and therefore can sense conformational changes in proteins. We have investigated changes in the radial coordinates of fluorescent probes bound to Cys-374 of F-actin when either of the isozymes (S1A1 and S1A2) of myosin subfragment 1 (S-1) bind. Using 5-[[2-[(iodoacetyl)amino]ethyl]amino]naphthalene 1-sulfonic acid and N-(4-dimethylamino-3,5-dinitrophenyl)maleimide as donor and acceptor probes, respectively, we calculated a radius of 13-14 A. This distance increased by approximately 4.5 A upon addition of S-1. No differences were detected between the effects of S1A1 and S1A2. This increase is reversed by MgATP. The average position of the probes on Cys-374 is closer to the filament axis than expected from the current models of F-actin. S-1 increases the radial position of Cys-374 either by direct interaction or via an allosteric conformational change associated with its binding. PMID- 9200684 TI - Cardiac muscarinic receptors. Cooperativity as the basis for multiple states of affinity. AB - Cooperativity has been investigated as the mechanistic basis for effects observed with cardiac muscarinic receptors in washed membranes from Syrian hamsters. Specifically, N-[3H]methylscopolamine labeled only 66-75% of the sites labeled by [3H]quinuclidinylbenzilate at apparently saturating concentrations of each radioligand. Also, receptors labeled by N-[3H]methylscopolamine revealed three states of affinity for agonists, both in native membranes and following irreversible blockade of about 80% of the sites by propylbenzilylcholine mustard; in both preparations, guanylylimidodiphosphate (GMP-PNP) effected an apparent interconversion of sites from higher to lower affinity for agonists and from lower to higher affinity for the antagonist. Excellent and mechanistically consistent descriptions of the data were obtained in terms of a model comprising cooperative and noncooperative forms of the receptor; the former was described by a variant of the Adair equation, and the latter was included to account for low affinity sites that survived treatment with the mustard. If differences in apparent capacity derive from negative cooperativity in the binding of N [3H]methylscopolamine, the cooperative form of the receptor was at least trivalent in native membranes; otherwise, constraints imposed by the effects of GMP-PNP at the concentrations of radioligand used in the assays dictate that the cooperative form of the receptor was at least tetravalent. In contrast, a divalent receptor is sufficient with the data from alkylated membranes, in accord with the reduced likelihood of interactions between functional sites within an oligomeric array. A model is presented wherein the receptor interconverts spontaneously between two or more states differing in their cooperative properties. The effects of GMP-PNP can be rationalized as a shift in the equilibrium between the different states. PMID- 9200685 TI - Cardiac muscarinic receptors. Relationship between the G protein and multiple states of affinity. AB - An expanded version of the mobile receptor model has been assessed in studies on the binding of N-[3H]methylscopolamine and [35S]GTPgammaS to cardiac muscarinic receptors and their attendant G proteins in ventricular membranes from hamster. The model comprises two pools of receptor, one of which lacks G proteins, and a heterogeneous population of G proteins that compete for the receptor within the G protein-containing pool. To guide the formulation of the model itself and to define the various parameters, data were combined from assays performed under various conditions with native membranes and following irreversible blockade of about 80% of the receptors with propylbenzilylcholine mustard. Multiple G proteins are indicated primarily by multiple states of affinity evident in the dose-dependent effect of guanyl nucleotides on the binding of carbachol; G protein-free receptors are indicated by sites of low affinity for carbachol that survive treatment with the mustard. The expanded model generally succeeds where more frugal schemes have been inadequate, but it nevertheless fails to yield a mechanistically consistent description of the data. Guanyl nucleotides and partial alkylation do not affect the inhibitory potency of carbachol in a manner consistent with their supposed effect on the equilibrium between uncoupled and G protein-coupled receptors. As inferred from the model, G proteins are lost upon alkylation of the receptor, and their numbers are regulated by guanyl nucleotides. Parameters estimated via N-[3H]methylscopolamine are wholly inconsistent with the same parameters estimated via [35S]GTPgammaS. The failure of the model suggests that multiple states of affinity may not arise from a ligand-regulated equilibrium between free receptors and G proteins on the one hand and one or more RG complexes on the other. PMID- 9200686 TI - Integrin alphaIIb beta3 reconstituted into lipid bilayers is nonclustered in its activated state but clusters after fibrinogen binding. AB - Integrin activation, ligand binding, and integrin clustering were analyzed using alphaIIb beta3 reconstituted into phospholipid vesicles and into supported planar lipid bilayers. Strong and specific binding of fibrinogen and the gamma-chain dodecapeptide of fibrinogen to alphaIIb beta3 indicated that the integrin is in an activated state after membrane reconstitution. Cryoelectron and fluorescence microscopy suggested a nonclustered state of the protein in the vesicle membrane. Supported planar lipid membranes were generated by fusion of vesicles in which approximately equal fractions of integrins were pointing inside-out and outside in. This distribution led to an immobilization of about 40% of the integrin in supported bilayers due to attachment of the large extracellular domains to the quartz support. Fluorescence recovery after photobleaching indicated a diffusion coefficient of D = (0.70 +/- 0.06) x 10(-8) cm2/s, consistent with a nonclustered state of the mobile integrin. Upon fibrinogen binding, the integrins became immobile, and fluorescence micrographs showed a patchy distribution of fibrinogen integrin complexes consisting of approximately 250 molecules. In addition to the expected dimer formation by bivalent fibrinogen, additionally induced fibrinogen clustering may account for the large size of the complexes. In contrast, binding of monovalent GRGDS pentapeptide or the gamma-chain dodecapeptide of fibrinogen altered neither the mobile fraction nor the association state of alphaIIb beta3. Our data indicate that integrin alphaIIbb3 is activated while monodisperse, and became clustered upon fibrinogen binding, leading to an irreversibly bound state. PMID- 9200687 TI - Spin-label studies on the anchoring and lipid-protein interactions of avidin with N-biotinylphosphatidylethanolamines in lipid bilayer membranes. AB - The specific binding of hen egg white avidin to phosphatidylcholine lipid membranes containing spin-labeled N-biotinylphosphatidylethanolamines (biotin PESLs) was investigated by using ESR spectroscopy. Spin-labeled biotin-PEs were prepared with the nitroxide group at position C-5, C-8, C-10, C-12, or C-14 of the sn-2 chain and were incorporated at 1 mol % in lipid bilayer membranes of dimyristoylphosphatidylcholine. Binding of avidin produced a strong and selective restriction of the biotin-PE lipid mobility at all positions of chain labeling, as shown by the ESR spectra recorded in the fluid lipid phase. The spectral components of the fraction of the biotin-PESLs that were not complexed by avidin indicated that the mobility of the bulk membrane lipids was unperturbed by binding avidin, as demonstrated by difference spectroscopy. Comparison of the positional profiles and temperature dependences of the outer hyperfine splittings from the biotin-PESLs suggests that the C-12 and C-14 positions of the avidin bound biotin-PEs are in register with the C-5 and C-7/C-6 positions, respectively, of the chains of the bulk membrane lipids. This indicates that the biotin-PEs are partially withdrawn from the membrane, with a vertical displacement of ca. 7-8 A, on complexation with avidin. In addition, the specific lipid-protein interaction with avidin results in a selective reduction in the rates of lipid chain motion, as shown by the increased ESR line widths. These data define the way in which avidin is anchored to lipid membranes containing biotin-PEs. PMID- 9200688 TI - Glucose transporter of Escherichia coli: NMR characterization of the phosphocysteine form of the IIB(Glc) domain and its binding interface with the IIA(Glc) subunit. AB - The transmembrane subunit of the glucose transporter, IICB(Glc), mediates vectorial transport with concomitant phosphorylation of glucose. Glucose phosphorylation proceeds through a cystein phosphate intermediate of the cytosolic IIB domain of IIC(Glc), which is phosphorylated by the IIA(Glc) subunit of the glucose transporter. Two- and three-dimensional NMR experiments were used to characterize the phosphorylation of the 10 kDa subclonal IIB domain and the complementary binding interfaces of [15N]IIB and [15N]IIA(Glc). The largest chemical shift perturbations and the only NOE differences accompanying IIB phosphorylation are confined to the active site residue Cys35, as well as Ile36, Thr37, Arg38, Leu39, and Arg40, which are all located in the turn between strands beta1 and beta2 and on beta2 itself. The significant increase of the amide cross peak intensities of Ile36, Thr37, and Arg38 upon phosphorylation suggests that the conformational freedom of these groups becomes restrained, possibly due to hydrogen bonding to the oxygens of the bound phosphate and to interactions between the guanidinium group of Arg38 and the phosphoryl group. The residues of IIB which experience chemical shift perturbations upon binding of IIA are located on a protruding surface formed by residues of strands beta1, beta2, and beta4, the beta4/alpha3 loop, and residues from the first two turns of alpha3. The corresponding binding surface of the IIA(Glc) domain is comprised of residues on five adjacent beta-strands and two short helices surrounding the active site His90. The binding surface of IIA(Glc) for IIB coincides with the binding surface for HPr, the phosphoryl carrier protein by which IIA(Glc) is phosphorylated [Chen, Y., Reizer, J., Saier, M. H., Fairbrother, W. J., & Wright, P. E. (1993) Biochemistry 32, 32-37]. PMID- 9200689 TI - Mutagenesis of the L7 loop connecting beta strands 12 and 13 of calcineurin: evidence for a structural role in activity changes. AB - Calcineurin is a heterodimer consisting of a catalytic A-subunit and a B-subunit, and is regulated by binding of calmodulin and calcium. The C-terminus of the A subunit contains an autoinhibitory domain which plays an important role in regulation of calcineurin activity. In this study, we have mutated the L7 loop connecting beta strands 12 and 13 in calcineurin A. These mutants included two chimeric mutants in which a four amino acid stretch in the cognate L7 loops of the related proteins phosphatase-1 (GEFD) or -2A (YRCG) were substituted for the calcineurin sequence DVYN (313-316), a point mutation (L312C), and a truncated mutant in which the YRG sequence replaced residues 313-316. Examination of the activities of these mutants led to the striking finding that truncation of the loop region by one residue resulted in hyperactivation of the calcineurin A subunit. That the hyperactivation is due to conformational effects on the catalytic core of the enzyme was established since this effect was maintained in truncation mutants (at residues 456 and 388) in which the calmodulin and autoinhibitory domains were deleted. These studies provide evidence that the L7 loop is an important structural element in the conformation of the active site, and may participate in the conformational transitions of calcineurin between a catalytically repressed state and an activated state under the influence of the B subunit. PMID- 9200690 TI - Insertion of Argos sequences into the B-loop of epidermal growth factor results in a low-affinity ligand with strong agonistic activity. AB - Recently, it has been shown that the activation of the Drosophila EGF receptor (DER) by its natural ligand Spitz is inhibited by Argos [Schweitzer, R., et al. (1995) Nature 376, 699-702]. Argos and Spitz both have an EGF-like domain which in the case of Argos differs from that of Spitz and other EGF receptor agonists in that it has an extended B-loop of 20 amino acids instead of 10 amino acids which in addition contains an unusual cluster of charged residues. To investigate whether B-loop sequences are an important determinant for receptor activation and play a causal role in the antagonistic activity of Argos, three human (h)EGF mutants were constructed in which amino acids derived from the Argos B-loop were introduced. In one mutant (E3A4E/B10), replacement of four amino acids in the B loop of hEGF (123, E24, D27, and K28) by the corresponding Argos residues neither altered the binding affinity of the growth factor for the hEGF receptor nor did it change its ability to induce a mitogenic response. Insertion of 2 additional Argos residues (E3A4E/B12) or extension of the B-loop by 10 amino acids (E3A4E/B20) resulted, however, in a significant loss of binding affinity. In spite of this, both E3A4E/B12 and E3A4E/B20 appeared to be strong agonists for the hEGF receptor with similar dose-response curves for mitogenic activity and MAPK activation as wild-type hEGF. These data show that several nonconservative substitutions in the hEGF B-loop are tolerated without affecting receptor binding or activation. Furthermore, they show that receptor binding and receptor signaling efficiency can be uncoupled which is a prerequisite for the development of receptor antagonists. PMID- 9200691 TI - Roles of the structure and orientation of ligands and ligand mimics inside the ligand-binding pocket of the vitamin D-binding protein. AB - 1alpha,25-Dihydroxyvitamin D3, the vitamin D hormone, manifests its diverse biological properties by specifically binding to the vitamin D sterol-binding pockets of vitamin D-binding protein (DBP) and vitamin D receptor. In the past, several affinity, photoaffinity, and chemical modification studies have been carried out to probe the vitamin D sterol-binding pocket of DBP and to evaluate the relationship between the structure of this pocket and the functions of the protein. In the present study, we examined the steric requirements inside this pocket by considering conformational structures of various bromoacetate derivatives of 25-hydroxyvitamin D3 and 1alpha,25-dihydroxyvitamin D3 and their abilities to covalently and specifically modify this pocket. We observed that, although 25-hydroxyvitamin D3 3beta-bromoacetate (25-OH-D3-3-BE), 1alpha,25 dihydroxyvitamin D3 3beta-bromoacetate [1alpha,25(OH)2D3-3-BE], 1alpha,25 dihydroxyvitamin D3 1alpha-bromoacetate [1alpha,25(OH)2D3-1-BE], and 1alpha,25 dihydroxyvitamin D3 1alpha,3beta-dibromoacetate [1alpha,25(OH)2D3-1,3-di-BE] bound DBP in a specific manner, only [3H]-25-OH-D3-3-BE and [3H]-1alpha,25(OH)2D3 3-BE affinity labeled the protein. BNPS-skatole cleavages of [3H]-25-OH-D3-3-BE- and 3H-1alpha,25(OH)2D3-3-BE-labeled DBP samples produced the same labeled peptide (N-terminal), demonstrating the specificity of labeling by these analogs. Energy-minimized conformational structures of these bromoacetate derivatives indicated significant changes in the A-ring conformations of these analogs. These structural changes were invoked to explain the inability of [3H]-1alpha,25(OH)2D3 1-BE and [3H]-1alpha,25(OH)2D3-1,3-di-BE to affinity label DBP. Overall, these studies suggested that the vitamin D sterol-binding pocket in DBP is sterically quite restrictive. This information could be potentially important in designing future vitamin D-based drugs for several diseases. PMID- 9200692 TI - Role of Leu99 of thrombin in determining the P2 specificity of serpins. AB - A recent study indicated that Tyr99 (chymotrypsin numbering) of factor Xa and Thr99 of activated protein C are S2 subsite residues that determine the P2 specificity of their substrates and inhibitors. To investigate the contribution of Leu99 to the P2 binding specificity of thrombin, three mutants of thrombin were prepared in which Leu99 was substituted with Tyr (L99Y), Thr (L99T), or Gly (L99G). Kinetic analysis indicated that antithrombin (AT with P2 Gly) inhibited thrombin L99Y, 14.1- and 5.5-fold slower than thrombin in the absence and presence of heparin, respectively. The L99Y mutation increased the stoichiometry of AT inhibition in the presence of heparin from approximately 1.6 to approximately 4.6, indicating that L99Y recognized AT as a substrate. The inhibition rates of L99T and L99G by AT, respectively, were 500.0- and 916.7-fold slower than thrombin in the absence of heparin but only 41.8- and 64.5-fold slower than thrombin in the presence of heparin. Resolution of the two-step reactions of AT with the mutant thrombins revealed that the impaired reactivities occurred in the second reaction step in which a non-covalent AT-thrombin encounter complex is converted to a stable, covalent complex. In reactions with protein C inhibitor (PCI with P2 Phe), L99Y was inhibited 3.5-fold slower than thrombin, L99T was inhibited at a similar or faster rate, and L99G was inhibited 23.9-fold faster than thrombin. The epidermal growth factor-like domains 4-6 of thrombomodulin (TM4-6) accelerated the PCI inhibition of wild-type and L99G thrombins 73.9- and 5.3-fold, respectively. Further studies indicated that the fibrinogen clotting and protein C activation rates by the mutants were impaired, but the cofactor function of TM was not affected as TM4-6 bound to wild-type [Kd(app) = 5.9 nM] and mutant thrombins with similar affinities [Kd(app) = 4.4 6.9 nM] and enhanced protein C activation rates by all mutants effectively. These results indicate that (1) Leu99 of thrombin is critical for determination of the P2 specificity of serpins, AT and PCI, (2) increasing the polarity of the S2 pocket of thrombin by introduction of a hydrophilic residue into this pocket is detrimental for reaction with AT, but it is tolerated in reaction with PCI, so that only the size of the S2 pocket of thrombin determines the P2 specificity of PCI, and (3) the thrombomodulin-induced conformational change that results in acceleration of thrombin inhibition by PCI involves Leu99. PMID- 9200693 TI - Evidence supporting a role for microfilaments in regulating the coupling between poorly dissociable IgE-Fc epsilonRI aggregates downstream signaling pathways. AB - Aggregation of Fc epsilonRI, the high-affinity receptor for IgE, on RBL-2H3 mast cells caused by reversible ligands such as multivalent antigen causes cellular responses that can be halted by subsequent addition of excess monovalent ligand. In contrast, Ca2+ and degranulation responses elicited by effectively irreversible streptavidin cross-linking of biotinylated IgE-Fc epsilonRI are not stopped by addition of excess biotin after stimulation is initiated. These results support previous conclusions based on studies with covalent oligomers of IgE that stable cross-links can continue to deliver stimulatory signals for extended periods of time. Dissociation measured in the presence of monovalent hapten reveals two populations of IgE-Fc epsilonRI cross-linked by multivalent antigen that differ in functional effectiveness. Aggregates with readily dissociable cross-links are normally responsible for triggering essentially all of the degranulation response, whereas aggregates with poorly dissociable cross links apparently do not trigger this response. Treatment of RBL-2H3 cells with cytochalasin D, an inhibitor of actin polymerization, enhances downstream signaling and enables the less readily dissociable aggregates to stimulate Ca2+ and degranulation responses. Under these conditions, cytochalasin D does not affect hapten-mediated dissociation of multivalent antigen, nor does it prevent hapten from reversing tyrosine phosphorylation of Syk. Cytochalasin D alone causes tyrosine phosphorylation of a protein at approximately 75 kDa, and it reduces hapten-induced reversal of antigen-stimulated tyrosine phosphorylation of several other proteins. Taken together, these results indicate that stimulated actin polymerization normally regulates the coupling of aggregated Fc epsilonRI to downstream signaling pathways, and they provide an explanation for seeming discrepancies between responses to stable and reversible cross-links. PMID- 9200694 TI - Detection of antigen-antibody binding events with the atomic force microscope. AB - An atomic force microscope (AFM) has been used to directly monitor specific interactions between antibodies and antigens employed in an immunoassay system. Results were achieved using AFM probes functionalized with ferritin, and monitoring the adhesive forces between the probe and anti-ferritin antibody coated substrates. Analysis of the force distribution data suggests a quantization of the forces, with a period of 49 +/- 10 pN. This periodic force may be attributed to single unbinding events between individual antigen and antibody molecules. These results demonstrate that the AFM could be employed as an analytical tool to study the interactions between the molecules involved in biosensor systems. The potential of the technique to provide information relating to the manner in which the antibody molecule binds to its specific antigen is also discussed. PMID- 9200695 TI - Potent inhibition of terminal complement assembly by clusterin: characterization of its impact on C9 polymerization. AB - The interactions of the heterodimeric apolipoprotein and complement inhibitor, clusterin (CL, 80 kDa), with actively assembling terminal complement proteins were characterized. Clusterin inhibited at three sites and by two modes of action. Clusterin inhibited C9 assembly on C5b-8 and C5b-9 and also bound to C5b 7 to prevent membrane attachment. The impact on C5b-9 assembly was the most potent. C9 assembly was monitored by assembly-induced fluorescence changes of C9 labeled with fluorescein isothiocyanate (FITC-C9). Assembly of monomeric FITC-C9 with C5b-8 or C5b-9(1) produced a substantial decrease in fluorescence intensity due to changes in the environment of the probe. Addition of the next subunit of unlabeled C9 produced a further small change. One equivalent of FITC-C9 bound to C5b-8 at low temperatures, but the fluorescence change and addition of more C9 did not occur until the temperaure was increased. Kinetic analysis of the fluorescence change suggested an irreversible, first-order process with an activation energy of 29 kcal/mol (k = 0.12 s(-1) at 25 degrees C). The kinetic properties differed for C9 addition to C5b-9(1) (0.27 s(-1) at 25 degrees C, 21 kcal/mol), indicating that C9 activation occurred at a different or altered site. Clusterin binding to C5b-8-(FITC-C9)1 caused fluorescence quenching similar to that of unlabeled C9, indicating that it bound to the C9 binding site. Clusterin binding to C5b-8 and C5b-9(1) was reversible with affinities that were 2 and 15 times that of C9 for the C5b-8 and C5b-9(1) complexes, respectively. The results suggested that the presence of <10% of the circulating clusterin in its heterodimeric, active form could reduce the rate of complement cytolysis of nucleated cells by 10-fold, and under some conditions by 100-fold or more. This would provide a high level of protection for certain cells and may allow time for action by other inhibitors of complement. PMID- 9200696 TI - Activation of the leukocyte NADPH oxidase subunit p47phox by protein kinase C. A phosphorylation-dependent change in the conformation of the C-terminal end of p47phox. AB - The leukocyte NADPH oxidase of neutrophils is a membrane-bound enzyme that catalyzes the production of O2- from oxygen using NADPH as electron donor. Dormant in resting neutrophils, the enzyme acquires catalytic activity when the cells are exposed to appropriate stimuli. During activation, the cytosolic oxidase components p47phox and p67phox migrate to the plasma membrane, where they associate with cytochrome b558, a membrane-bound flavohemoprotein, to assemble the active oxidase. An essential element of the activation process is the phosphorylation of p47phox, an event that accompanies oxidase activation in whole cells and can activate the oxidase in a cell-free system. We show here that the phosphorylation of p47phox leads to a substantial decrease in the reactivity of cysteine C378 toward N-ethylmaleimide, indicating the occurrence of a conformational change involving the C-terminal region of p47phox. A similar conformational change occurs when p47phox is exposed to arachidonate, one of a number of anionic detergents that activate the oxidase in the cell-free system. We propose that this change in conformation results in the appearance of a binding site through which p47phox interacts with cytochrome b558 during the activation process. PMID- 9200697 TI - Structure of glycan moieties responsible for the extended circulatory life time of fetal bovine serum acetylcholinesterase and equine serum butyrylcholinesterase. AB - Cholinesterases are serine hydrolases that can potentially be used as pretreatment drugs for organophosphate toxicity, as drugs to alleviate succinylcholine-induced apnea, and as detoxification agents for environmental toxins such as heroin and cocaine. The successful application of serum-derived cholinesterases as bioscavengers stems from their relatively long residence time in the circulation. To better understand the relationship between carbohydrate structure and the stability of cholinesterases in circulation, we determined the monosaccharide composition, the distribution of various oligosaccharides, and the structure of the major asparagine-linked oligosaccharides units present in fetal bovine serum acetylcholinesterase and equine serum butyrylcholinesterase. Our findings indicate that 70-80% of the oligosaccharides in both enzymes are negatively charged. This finding together with the molar ratio of galactose to sialic acid clearly suggests that the beta-galactose residues are only partially capped with sialic acid, yet they displayed a long duration in circulation. The structures of the two major oligosaccharides from fetal bovine serum acetylcholinesterase and one major oligosaccharide from equine serum butyrylcholinesterase were determined. The three carbohydrate structures were of the biantennary complex type, but only the ones from fetal bovine serum acetylcholinesterase were fucosylated on the innermost N-acetylglucosamine residue of the core. Pharmacokinetic studies with native, desialylated, and deglycosylated forms of both enzymes indicate that the microheterogeneity in carbohydrate structure may be responsible, in part, for the multiphasic clearance of cholinesterases from the circulation of mice. PMID- 9200699 TI - Pigment assignment in the absorption spectrum of the photosystem II reaction center by site-selection fluorescence spectroscopy. AB - The steady state fluorescence properties of the photosystem II reaction center (D1-D2-cyt-b559 complex, PSII-RC) have been investigated by site-selection spectroscopy. The pattern of the vibronic bands in the emission spectra is used to identify the fluorescing species that have their absorption maxima on the red edge of the spectrum (at around 682 nm). At 10 K, even samples with a low content of red absorbing chlorophyll a (Chl) show pure Chl emission upon excitation at 685 nm, whereas at 77 K the fluorescence of the PSII-RCs is contributed to by Chl and pheophytin a (Pheo) in a ratio of roughly 8:2. These results allow an unequivocal distinction between two different spectral decompositions that were recently suggested for the absorption spectrum of the PSII-RC [Konermann, L., & Holzwarth, A. R. (1996) Biochemistry 35, 829]. Only one of these decompositions is compatible with the experimental data presented here according to which the absorption on the red edge of the spectrum is dominated by an accessory Chl. PMID- 9200698 TI - Threonine-89 participates in the active site of bacteriorhodopsin: evidence for a role in color regulation and Schiff base proton transfer. AB - Bacteriorhodopsin (bR) functions as a light-driven proton pump in the purple membrane of Halobacterium salinarium. A major feature of bR is the existence of an active site which includes a retinylidene Schiff base and amino acid residues Asp-85, Asp-212, and Arg-82. This active site participates in proton transfers and regulates the visible absorption of bacteriorhodopsin and its photointermediates. In this work we find evidence that Thr-89 also participates in this active site. The substitution Thr-89 --> Asn (T89N) results in changes in the properties of the all-trans retinylidene chromophore of light-adapted bR including a redshift of the visible lambda(max) and a downshift in C=N and C=C stretch frequencies. Changes are also found in the M and N intermediates of the T89N photocycle including shifts in lambda(max), a downshift of the Asp-85 carboxylic acid C=O stretch frequency by 10 cm(-1), and a 3-5-fold decrease in the rate of formation of the M intermediate. In contrast, the properties of the 13-cis retinylidene chromophore of dark-adapted T89N as well as the K and L intermediates of the T89N photocycle are similar to the wild-type bacteriorhodopsin. These results are consistent with an interaction of the hydroxyl group of Thr-89 with the protonated Schiff base of light-adapted bR and possibly the N intermediate but not the 13-cis chromophore of dark-adapted bR or the K and L intermediates. Thr-89 also appears to influence the rate of Schiff base proton transfer to Asp-85 during formation of the M intermediate, possibly through an interaction with Asp-85. In contrast, the hydroxyl group of Thr-89 is not obligatory for proton transfer from Asp-96 to the Schiff base during formation of the N intermediate. PMID- 9200701 TI - Characterization of pheophytin ground states in Rhodobacter sphaeroides R26 photosynthetic reaction centers from multispin pheophytin enrichment and 2-D 13C MAS NMR dipolar correlation spectroscopy. AB - The electronic ground states of pheophytin cofactors potentially involved in symmetry breaking between the A and B branch for electron transport in the bacterial photosynthetic reaction center have been investigated through a characterization of the electron densities at individual atomic positions of pheophytin a from 13C chemical shift data. A new experimental approach involving multispin 13C labeling and 2-D NMR is presented. Bacterial photosynthetic reaction centers of Rhodobacter sphaeroides R26 were reconstituted with uniformly 13C biosynthetically labeled (plant) Pheo a in the two pheophytin binding sites. From the multispin labeled samples 1-D and 2-D solid-state 13C magic angle spinning NMR spectra could be obtained and used to characterize the pheophytin a ground state in the Rb. sphaeroides R26 RCs, i.e., without a necessity for time consuming selective labeling strategies involving organic synthesis. From the 2-D solid state 13C-13C correlation spectra collected with spinning speeds of 8 and 10 kHz, with mixing times of 1 and 0.8 ms, many 13C resonances of the [U-13C]Pheo a molecules reconstituted in the RCs could be assigned in a single set of experiments. Parts of the pheophytins interacting with the protein, at the level of 13C shifts modified by binding, could be identified. Small reconstitution shifts are detected for the 17(2) side chain of ring IV. In contrast, there is no evidence for electrostatic differences between the two Pheo a, for instance, due to a possibly strong selective electrostatic interaction with Glu L104 on the active branch. The protonation states appear the same, and the NMR suggests a strong overall similarity between the ground states of the two Pheo a, which is of interest in view of the asymmetry of the electron transfer. PMID- 9200700 TI - Quenching of chlorophyll a fluorescence in the aggregates of LHCII: steady state fluorescence and picosecond relaxation kinetics. AB - The protein composition, steady state and time-resolved fluorescence emission spectra were studied in solubilized and aggregated LHCII complexes, that were prepared according to two different isolation protocols: (1) by fractionation of cation-depleted thylakoid membranes using the non-ionic detergent Triton X-100 according to the procedure of Burke et al. [(1978) Arch. Biochem. Biophys. 187, 252-263] or (2) by solubilization with N-beta-dodecyl maltoside (beta-DM) of photosystem II (PSII) membrane fragments in the presence of cations [Irrgang et al. (1988) Eur. J. Biochem. 178, 207-217]. Based on the analysis of the decay associated emission spectra measured at 10 and 80 K five long-wavelength chlorophyll species were identified in aggregated LHCII complexes. These five forms are characterized by emission maxima at 681.5, 683, 687, 695, or 702 nm. All of these forms were found in both types of LHCII preparations but the relative amounts and temperature dependency of these species were markedly different in the aggregated LHCII complexes isolated by the two procedures. It was found that these differences cannot be simply explained by effects due to using a less mild detergent as beta-DM or by an ionic influence of Ca2+. Biochemical analysis of the protein composition showed that beta-DM type LHCII consists of all the chlorophyll (Chl)binding proteins belonging to the antenna system of PSII except the CP29 type II gene product (CP29). In contrast, the Triton X-100-solubilized LHCII is highly depleted in CP26 (CP 29 type I gene product) and is contaminated by a variety of unidentified polypeptides. It is proposed that the aggregates of LHCII prepared using Triton X-100 acquire specific spectral and kinetic features due to interaction between the bulk of LHCII subunits and minor protein(s). PMID- 9200702 TI - Parallel polarization electron paramagnetic resonance studies of the S1-state manganese cluster in the photosynthetic oxygen-evolving system. AB - Magnetic properties of the S1-state manganese cluster in the oxygen-evolving photosystem II were studied by parallel polarization electron paramagnetic resonance spectroscopy. Dark minus light spectra gave rise to a broad S1-state signal with a g value of about 4.9 [Dexheimer, S. L., Klein, M. P. (1992) J. Am. Chem. Soc. 114, 2821-2826]. Temperature variation of the signal intensity between 1.9 and 10 K observed in PS II with a sucrose buffer indicates that the signal originates from an excited state with a spin S of 1 with separation from the ground state (S = 0) of about 2.5 K. The S1-state signal was also observed in the sucrose buffer supplemented by 50% glycerol. However, no S1-state signal was detected by addition of 3% methanol or 30% ethylene glycol in the sucrose buffer, although illumination at 200 K in the presence of these alcohols induced the normal multiline S2 signal. Furthermore, modification of the Mn cluster by Cl- or Ca2+ depletion from PS II membranes failed to produce a detectable S1-state signal. A possible magnetic structure of the Mn cluster responsible for the generation of the S1-state signal is discussed on the basis of these observations. PMID- 9200703 TI - Detection of an Fe2+-protoporphyrin-IX intermediate during aspirin-treated prostaglandin H2 synthase II catalysis of arachidonic acid to 15-HETE. AB - Spectral intermediates associated with the dioxygenase and peroxidase activities of prostaglandin H2 (PGH2) synthase I and II were monitored by stopped-flow spectrometry. During reactions of PGH2 synthase I with arachidonic acid (AA) and ethyl hydrogen peroxide (EtOOH), compound I (Fe5+; formally (protoporphyrin-IX) x +Fe4+=O) and compound II (Fe4+; formally (protoporphyrin-IX)Fe4+=O) were detected. These intermediates were observed sooner with EtOOH (within 50 ms) than with AA (within 200 ms). Compound I and compound II were found to be kinetically competent with respect to AA-dependent O2 uptake. These findings are consistent with a mechanism in which peroxidative cleavage precedes AA dioxygenation. During reactions with PGH2 synthase II with AA, compound I and compound II were again observed within 200 ms and were kinetically competent to participate in dioxygenation. However, during reactions of PGH2 synthase II with EtOOH, compound I and compound II were detected much later (after 10 s). These findings would be inconsistent with a mechanism in which peroxidative cleavage precedes AA dioxygenation. When aspirin-treated PGH2 synthase II was reacted with EtOOH, a normal peroxidase cycle occurred with compound I and compound II formation occurring over 10 s. However, when aspirin-treated PGH2 synthase II was reacted with AA, a unique spectral intermediate with lambda(max) at 446 nm was detected within 3 ms and was strikingly similar to ferrous (Fe2+) protoporphyrin-IX. Aspirin-treated PGH2 synthase II was found to produce 15-HETE, and the appearance of the Fe2+ intermediate (within 3 ms) indicated that it was kinetically competent to participate in the 15-dioxygenation event. The detection of this Fe2+ intermediate and the slow formation of compound I and compound II observed with EtOOH in PGH2 synthase II suggest that peroxidative cleavage is not the initiating event in dioxygenation. Instead, it is proposed that the reduction of Fe3+ in heme to Fe2+ oxidizes a peroxide to yield an initiating peroxy radical. Since it is unlikely that 11- and 15-dioxygenation occurs via different mechanisms, our findings question mechanisms of catalysis in both PGH2 synthases. PMID- 9200704 TI - Function of conserved tryptophans in the Aspergillus niger glucoamylase 1 starch binding domain. AB - Nuclear magnetic resonance (NMR) and ultraviolet (UV) difference spectroscopy were used to assess the role of a number of tryptophan residues in the granular starch binding domain (SBD) of glucoamylase 1 from Aspergillus niger. Wild-type SBD and three variant (W563K, W590K, and W615K) proteins were produced using an A. niger expression system. Titration studies were conducted with beta cyclodextrin (betaCD), a cyclic analogue of starch, as the ligand. The NMR studies show that the W563K and W590K variants only bind 1 equiv while the wild type protein forms a 2:1 (ligand:protein) complex. It also clearly demonstrates the abolition of binding at site 1 and site 2 in W590K and W563K, respectively. UV difference spectroscopy was used to calculate dissociation constants with addition of betaCD: 14.4 microM (apparent) for the wild type, 28.0 microM for W563K, and 6.4 microM for W590K. The implication of this is that the two binding sites have unequal contributions to the overall binding of the SBD which may be related to functional differences between the two binding sites. The low stability of the third variant, W615K, suggests that this tryptophan is not involved in binding but has an essential structural role. PMID- 9200705 TI - Kinetic analysis of human deoxycytidine kinase with the true phosphate donor uridine triphosphate. AB - Deoxycytidine kinase is the rate-limiting process in the activation for several clinically important antitumor agents. Previous studies have focused on deoxycytidine (dCyd) and adenosine triphosphate (ATP) as substrates for this enzyme. In view of recent data indicating that uridine triphosphate (UTP) is the physiologic phosphate donor for this enzyme, a study of the kinetic properties of dCyd kinase with dCyd and UTP was undertaken. The results presented here demonstrate that UTP and ATP produce kinetically distinguishable differences in nucleoside phosphorylation by dCyd kinase. At high dCyd concentrations, dCyd kinase exhibited substrate activation with ATP. In contrast, in the presence of UTP, substrate inhibition was observed at concentrations of dCyd greater than 3 microM. Inhibition by dCyd was noncompetitive with respect to UTP and could not be reversed by a 200-fold increase in UTP concentration, indicating that the inhibition was not due to dCyd binding at the nucleotide binding site. The kinetic mechanism for dCyd kinase was determined with dCyd and UTP as substrates. UTP was the preferred phosphate donor with a true Km value of 1 microM compared to 54 microM with ATP, resulting in a 50-fold greater substrate efficiency for UTP. Although the double-reciprocal plots with UTP produced parallel lines, initial velocity plots with other phosphate donors and product inhibition studies indicated that dCyd kinase formed a ternary complex with its substrates. The parallel lines with UTP were apparently due to a low dissociation constant for UTP, which was calculated as more than 13-fold lower than its Km value. Analysis of product inhibition studies indicated that dCyd kinase followed an ordered A-B random P-Q reaction sequence, with UTP as the first substrate to bind. In contrast, previous results demonstrated a random bi-bi sequence for dCyd kinase in the presence of ATP. The combined results indicate that the enzyme can follow a random bi-bi reaction sequence, but with UTP as the phosphate donor, the addition of nucleotide prior to dCyd is strongly preferred. The noncompetitive substrate inhibition, which was independent of UTP concentration, indicates that high concentrations of dCyd promote addition of the nucleoside prior to UTP, resulting in a lower velocity. PMID- 9200706 TI - Electrostatic effects on substrate activation in para-hydroxybenzoate hydroxylase: studies of the mutant lysine 297 methionine. AB - p-Hydroxybenzoate hydroxylase (EC 1.14.13.2) is a flavoprotein monooxygenase that catalyzes the incorporation of one atom of molecular oxygen into p hydroxybenzoate to form 3,4-dihydroxybenzoate. The enzyme activates the substrate at the 3 position to electrophilic substitution by lowering the pKa of the phenolic oxygen. The results presented here indicate that regions of positive potential in the active site facilitate this substrate activation, which is necessary for rapid hydroxylation. We have neutralized a positive point charge by mutating lysine 297 to methionine (K297M). This mutation changes an amino acid near the active site, but not directly in contact with the flavin or the substrate. A variety of transient state kinetic and static parameters have been determined with two substrates. The results indicate that the K297M mutant does not activate the substrate through phenolic ionization to the same extent as wild type (WT) and yet remains a competent hydroxylase. However, catalysis by the mutant is slow compared to that of WT, particularly in the oxidative half reaction. Thus, normally quite labile oxygenated flavin intermediates encountered in the hydroxylation pathway of WT p-hydroxybenzoate hydroxylase are stabilized and their decay is rate limiting in the K297M turnover. Electrostatic potential calculations offer an explanation for the lack of substrate activation. The stability of the oxidative reaction intermediates seems to be related to a lower degree of substrate activation. PMID- 9200707 TI - Reconstitution of human base excision repair with purified proteins. AB - Base excision repair is a major mechanism for correcting aberrant DNA bases. We are using an in vitro base excision repair assay to fractionate and purify proteins from a human cell extract that are involved in this type of repair. Three fractions are required to reconstitute base excision repair synthesis using a uracil-containing DNA as a model substrate. We previously showed that one fraction corresponds to DNA polymerase beta. A second fraction was extensively purified and found to possess uracil-DNA glycosylase activity and was identified as the product of the UNG gene. A neutralizing antibody to the human UNG protein inhibited base excision repair in crude extract by at least 90%. The third fraction was highly purified and exhibited apurinic/apyrimidinic (AP) endonuclease activity. Immunoblot analysis identified HAP1 as the major polypeptide in fractions possessing DNA repair activity. Recombinant versions of UNG, HAP1, and DNA polymerase beta were able to substitute for the proteins purified from human cells. Addition of DNA ligase I led to ligated repair products. Thus, complete base excision repair of uracil-containing DNA was achieved by a combination of UNG, HAP1, DNA polymerase beta, and DNA ligase I. This is the first complete reconstitution of base excision repair using entirely eukaryotic proteins. PMID- 9200708 TI - Rapid-reaction analysis of plasmid DNA cleavage by the EcoRV restriction endonuclease. AB - Rapid-reaction methods have been used previously to identify intermediates in the reaction of the EcoRV restriction endonuclease on oligonucleotide substrates. In this study, the pathway on macromolecular DNA was elucidated by using the quench flow method to analyze EcoRV reactions on a plasmid with one recognition site. Some reactions were carried out by first allowing the EcoRV enzyme to bind nonspecifically to the DNA and then initiating DNA cleavage by adding magnesium ions. The subsequent transfer of the enzyme from nonspecific to specific sites was extremely rapid, at a random walk rate of at least 5 x 10(5) base pairs per second. The two strands of the DNA at the EcoRV recognition site were then cleaved sequentially, at rates that were faster than the turnover number of the enzyme. The rates recorded for the cleavage steps were direct measurements of phosphodiester hydrolysis, while the turnover is limited by the dissociation of the product cleaved in both strands. Other reactions were initiated by adding EcoRV and MgCl2 to the DNA: these revealed not only the processes observed in reactions starting from DNA-bound enzyme but also the bimolecular association of the protein with the plasmid. The association rate was limited by diffusion but its rate constant, 1.2 x 10(8) M(-1) s(-1), was unusually small for the binding of a protein to DNA. The slowness of this diffusion-controlled process may be due to a rapid oscillation of the protein between closed and open conformations, with only the open form capable of binding DNA. PMID- 9200709 TI - Kinetic analysis of a mutational hot spot in the EcoRV restriction endonuclease. AB - The EcoRV endonuclease contacts the minor groove of DNA through a peptide loop encompassing residues 67-72. This loop adapts to distorted DNA in the specific complex and to regular DNA in the nonspecific complex. Random mutagenesis had previously identified glutamine 69 as the key component of the loop and this study reports on mutants with glutamate (Q69E), lysine (Q69K), or leucine (Q69L) at this position. The mutants bound DNA specifically at the EcoRV recognition site in the presence of Ca2+, in the same manner as wild-type EcoRV. In the absence of divalent metals, Q69K and Q69L showed the same nonspecific binding as native EcoRV while Q69E failed to bind DNA. Glutamate at position 69 presumably repels nonspecific DNA whilst allowing the adaptations to specific DNA. Both Q69E and Q69K had severely impaired DNA cleavage activities, while Q69L had a steady state k(cat) within an order of magnitude of wild-type EcoRV though its primary product was nicked DNA, in contrast to double strand breaks by wild-type EcoRV. The activity of Q69L required higher concentrations of Mg2+ than the wild-type and showed a sigmoidal dependence upon the Mg2+ concentration, indicating two metal ions per strand scission. Transient kinetics on Q69L gave lower rate constants for phosphodiester hydrolysis than wild-type EcoRV and its reaction also involved a slow conformational change preceding DNA cleavage that had no equivalent with the wild-type. Gln69 in EcoRV thus plays key roles in the adjustments of the protein to varied DNA structures and in the alignment of the catalytic functions for DNA cleavage. PMID- 9200710 TI - Mechanism of DNA-dependent protein kinase inhibition by cis diamminedichloroplatinum(II)-damaged DNA. AB - We have determined the mechanism of DNA-dependent protein kinase (DNA-PK) inhibition by cis-diamminedichloroplatinum(II)-(cisplatin-) damaged DNA. We previously have demonstrated that Ku, the DNA binding subunit of DNA-PK, is capable of binding to DNA duplexes globally damaged with cisplatin but was unable to stimulate DNA-PKcs, the catalytic subunit [Turchi & Henkels (1996) J. Biol. Chem. 271, 2992-3000]. In this report we have assessed Ku binding and DNA-PK stimulation using a series of DNA substrates containing single, site-specific d(GpG), d(ApG), and d(GpXpG) intrastrand cisplatin adducts and a substrate with a single interstrand cisplatin adduct. Results demonstrate that Ku binding is marginally decreased by the presence of cisplatin adducts on each substrate. When assayed for the ability to stimulate DNA-PK, each cisplatin-damaged substrate resulted in significantly decreased activity compared to undamaged DNA controls. The degree of inhibition of both Ku binding and kinase activity varied depending on the specific adduct employed. The inhibition of DNA-PK activity by cisplatin damaged DNA was observed using either a synthetic peptide or human replication protein A as a substrate. Autophosphorylation of the DNA-PKcs and Ku subunits was also inhibited in reactions performed with cisplatin-damaged DNA, demonstrating that increased autophosphorylation of DNA-PKcs does not account for the decreased kinase activity observed with cisplatin-damaged DNA. Equilibrium binding and initial velocity experiments revealed a less than 2-fold increase in the Kd of Ku and the Km of DNA-PK for DNA containing a single 1,2-d(GpG) cisplatin adduct. The mechanism of DNA-PK inhibition by cisplatin-damaged DNA can be attributed to a large decrease in the Vmax and small increase in Km. PMID- 9200711 TI - Evidence supporting a role for histidine-235 in cation binding to human 3-hydroxy 3-methyglutaryl-CoA lyase. AB - Histidine-235 of human 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) lyase is the second basic residue in a conserved HXH motif. This residue is solvent accessible, readily reacting with the group specific reagent diethyl pyrocarbonate. Site-directed mutagenesis has been employed to substitute alanine or aspartate for H235. Characterization of the isolated H235A and H235D lyase mutants indicates that their tertiary structure is substantially intact. The mutant proteins, like the wild-type enzyme, are stoichiometrically modified by the affinity label, 2-butynoyl-CoA. Catalytic activity of the mutants is diminished by 15-fold and Km for HMG-CoA elevated approximately 4-fold in comparison with the values for wild-type enzyme. The function of H235 is suggested by investigation of the interaction of these enzymes with the dissociable divalent cation (e.g. Mg2+ or Mn2+) that is required for activity. ESR experiments show that wild-type enzyme forms a stable binary E*M complex. In contrast, H235A and H235D proteins do not efficiently form a binary complex. Significant interaction with cation (Mn2+) only occurs in the presence of the substrate analog, 3-hydroxyglutaryl-CoA. Similarly, when cation interaction is estimated in the presence of substrate using steady-state kinetic approaches, activator constants (Ka) and divalent cation Km values are measurable but are elevated by 15-90-fold over comparable estimates for the wild-type enzyme. The data confirm our earlier suggestion that both protein and substrate contribute ligands to HMG-CoA lyase's divalent cation activator. More specifically, the current observations suggest that H235 has an important function in cation binding. PMID- 9200712 TI - Interactions of structural C and regulatory N domains of troponin C with repeated sequence motifs in troponin I. AB - The actomyosin ATPase inhibitory protein troponin I (TnI) plays a central regulatory role in skeletal and cardiac muscle contraction and relaxation through its calcium-dependent interactions with troponin C (TnC) and actin. Previously we have demonstrated the utility of F29W and F105W mutants of TnC for measurement of binding affinities of inhibitory peptide TnI(96-116) to its regulatory N and structural C domains, both in isolation and in the intact TnC molecule [Pearlstone, J. R. & Smillie, L. B. (1995) Biochemistry 34, 6932-6940]. This approach is now extended to fragment TnI(96-148). Curve-fitting analyses of fluorescence changes induced in the intact TnC mutants and the isolated N and C domains by increasing [TnI(96-148)] have permitted the assignments of K(D) values (designated K(D,N) and K(D,C)) to the interaction of TnI(96-148) with the N and C domains, respectively, of intact TnC. Taken together with the previous data for TnI(96-116) binding, it can be concluded that, within TnI(96-148), residues 96 116 are primarily responsible for binding to C domain of intact TnC and residues 117-148 to its N domain. Inspection of the available mammalian and avian skeletal muscle TnI amino acid sequences reveals a previously unrecognized conserved motif repeated 3-fold, once in the inhibitory peptide region (approximately residues 101-114; designated alpha) and twice more in the region of residues approximately 121-132 (beta) and approximately 135-146 (gamma). The number and distribution of these motifs have important structural implications for the TnI x C complex. In the beta motif of cardiac TnI, as compared with skeletal, several changes in charged amino acids are suggested as candidates responsible for the greater sensitivity of cardiac Ca2+-regulated actomyosin to acidic pH as in ischemia. PMID- 9200713 TI - Interaction of smooth muscle myosin phosphatase with phospholipids. AB - The 130 kDa myosin-binding subunit (MBS) of smooth muscle myosin phosphatase was detected in cytoskeletal, cytosolic, and membrane fractions of T24 cells. Also, MBS was distributed between cytoplasm and plasmalemma in mitotic REF52 cells. These observations prompted this study of the interaction(s) of phospholipids with myosin phosphatase. Using a sedimentation assay, gizzard myosin phosphatase bound to vesicles of acidic phospholipids, i.e. phosphatidylserine (PS), phosphatidylinositol, and phosphatidic acid (PA). Neutral phospholipids did not bind. Binding of PS to myosin phosphatase also was demonstrated by electrophoresis under nondenaturing conditions. Preferential binding of PA, compared to that of the other acidic phospholipids, was indicated. Interaction of acidic phospholipids with myosin phosphatase inhibited phosphatase activity toward phosphorylated myosin. The extent of PS binding with myosin phosphatase decreased on increasing ionic strength and Mg2+ concentration. MBS (M130/M133) and M20 were phosphorylated by protein kinase A to 3 and 1 mol of P/(mol of subunit), respectively. Phosphorylation of the holoenzyme decreased phospholipid binding with recovery of phosphatase activity. Using limited proteolysis of the holoenzyme and various mutants, it was shown that phospholipid binding was associated with the C-terminal part of MBS, Ser 667-Ile 1004, and M20. The phosphorylation site involved in regulation of phospholipid binding is within the C-terminal MBS sequence. These results suggest that myosin phosphatase may interact with membranes and that phosphorylation by protein kinase A could modify this interaction. This mechanism could be important in localization of myosin phosphatase and in targeting substrates at different loci. PMID- 9200714 TI - Structural analysis of apolipoprotein A-I: limited proteolysis of methionine reduced and -oxidized lipid-free and lipid-bound human apo A-I. AB - The domain structures of lipid-free and lipid-bound apolipoprotein A-I (apo A-I) containing reduced and oxidized methionines were analyzed by limited proteolysis. Lipid-free apo A-I is cleaved primarily in the extreme carboxy-terminus and, to a much lesser extent, in the central region of the protein between residues 115 and 136. Oxidation of methionines 112 and 148 to the corresponding sulfoxides in putative amphipathic helices 4 (P99-E120) and 6 (P143-A164), respectively, causes helices 1 (L44-G65), 2 (P66-S87), and 7 (P165-G186) to become susceptible to protease digestion. These results are consistent with a discrete, globular tertiary structure for the lipid-free protein minimally formed from amphipathic helices 1, 2, 4, 6, and 7. In distinct contrast to lipid-free apo A-I, lipid bound apo A-I is most susceptible to cleavage in the extreme amino-terminus and, to a lesser extent, in both the central and carboxy-terminal regions. The observed cleavage pattern for the reduced lipid-bound protein supports the existence of many of the turns between helices predicted by sequence analysis of the lipid-bound protein. Methionine oxidation of lipid-bound protein results in a decreased protease susceptibility in the extreme amino-terminus and a concomitant increase in protease susceptibility in the central and carboxy-terminal regions. The results from methionine oxidation indicate the oxidation state of the protein is an important determinant in defining the conformation of both lipid-free and lipid-bound apo A-I. PMID- 9200715 TI - Very high single channel water permeability of aquaporin-4 in baculovirus infected insect cells and liposomes reconstituted with purified aquaporin-4. AB - The insect cell/baculovirus system was used to express the mercurial-insensitive water channel aquaporin-4 (AQP4) for purification and reconstitution. Immunoblot analysis of Sf9 cells infected with recombinant baculovirus showed greatest AQP4 expression at 72 h after infection at a multiplicity-of-infection of 5. Immunostaining and cell membrane fractionation indicated AQP4 plasma membrane expression. Quantitative immunoblot analysis showed approximately 60 microg of AQP4 per milligram of plasma membrane protein (approximately 2 mg of AQP4 protein per liter of Sf9 cell culture). Functional analysis by stopped-flow light scattering indicated that AQP4 functioned as a mercurial-insensitive water selective transporter. Osmotic water permeability (Pf) in plasma membrane vesicles from AQP4-expressing Sf9 cells was very high (0.053 cm/s at 10 degrees C), weakly temperature dependent (activation energy, 4.5 kcal/mol), and not inhibited by HgCl2. The AQP4 single channel water permeability (p(f)), estimated from Pf and protein amount, was 19 x 10(-14) cm3/s. Purification of AQP4 to a single Coomassie blue-stained protein on SDS-PAGE (1300-fold over homogenate) was achieved by membrane fractionation, carbonate stripping of nonintegral proteins, solubilization in octyl-beta-glucoside, and anion exchange chromatography. AQP4 protein identity was confirmed by mass spectrometry. Reconstitution of purified AQP4 into proteoliposomes increased osmotic water permeability by >40-fold, giving a p(f) of 15 x 10(-14) cm3/s, remarkably greater than that of 4.9 x 10( 14) cm3/s measured in parallel for AQP1. These results establish the first purification of an aquaporin from a heterologous expression system. The high AQP4 p(f) suggests (a) significant functional differences among the aquaporins, (b) inadequacy of existing pore models to account for high water flow and water permselectivity, and (c) possible enhancement of water flow by AQP4 assembly in orthogonal arrays. PMID- 9200716 TI - Quantal analysis suggests presynaptic involvement in expression of neocortical short- and long-term depression. AB - Long-term depression together with long-term potentiation represent popular experimental models to study synaptic plasticity. However, analyses of the mechanisms underlying the expression of cortical long-term depression are in their infancy and have been confined to the hippocampus. Short- and long-term depression in neocortex is not well understood. Here we recorded small excitatory postsynaptic potentials intracellularly from rat visual cortex slices. The responses fluctuated between several amplitude levels suggesting a quantal nature of the synaptic transmission. Consistent changes in the quantal steps accompanied neither paired-pulse depression (50 ms interval within the pair) nor long-term depression (induced by 1 Hz, 5 min stimulation). The amplitude distributions shifted to smaller values suggesting decreases in the number of quanta released without essential changes in the postsynaptic quantal efficiency. Both the coefficient of variation of response amplitudes and the number of response failures increased; cases were encountered suggesting a very low release probability after depression. Changes in quantal content estimated from the deconvolution analysis were correlated with the magnitude of depression. The findings suggest predominantly presynaptic loci for expression of short- and long term neocortical depressions. The likely underlying mechanism is a decrease in transmitter release probability. Long-term depression decreased the probability so strongly that some inputs became virtually silent. PMID- 9200717 TI - Loss of autoreceptor function in dopaminergic neurons from dopamine D2 receptor deficient mice. AB - Dopamine plays a key role in the control of motor and cognitive functions through the interaction with membrane receptors. Dopamine elicits its physiological effect by interacting with receptors that belong to the seven transmembrane domain G-protein-coupled receptors family. Pharmacological and structural analyses have allowed the division of these receptors into two classes: the D1- and D2-like receptors. The D1-like subfamily comprises D1 and D5 while the D2 like is formed by D2, D3 and D4. Dopaminergic neurons arise from the ventral tegmental area and the substantia nigra. These neurons give rise to four dopaminergic pathways: the nigrostriatal, the mesolimbic, the mesocortical and tuberoinfundibular pathways. These pathways are involved in the control of movement, learning, motivation reward and hormone synthesis and release. Dysfunction in these pathways leads to neurological, psychiatric and endocrine disorders. Indeed, degeneration of the nigrostriatal pathway leads to Parkinson's disease in humans, characterized by a strong reduction of released dopamine. Thus, a fine tuning of the firing discharge of dopaminergic neurons is a key function in the regulation of dopamine mediated activities in the central nervous system. Somatodendritic dopaminergic autoreceptors of the D2-like family are responsible for such a function. However, it is still controversial whether this function could be ascribed only to one or more members of this subfamily. PMID- 9200718 TI - Synapsin I and syntaxin 1B: key elements in the control of neurotransmitter release are regulated by neuronal activation and long-term potentiation in vivo. AB - The messenger RNAs encoding proteins of the exocytotic machinery were measured at different times following the induction of long-term potentiation or increasing neuronal activity in the dentate gyrus of the rat in vivo. In situ hybridization revealed that from the many messenger RNAs that encode proteins involved in regulated exocytosis, only those encoding synapsin I and syntaxin 1B were specifically increased. The levels of messenger RNA encoding both synapsin I and syntaxin 1B were increased on the ipsilateral side of the dorsal dentate gyrus 2 and 5 h following the induction of long-term potentiation. Syntaxin 1B was also increased in the ventral dentate gyrus at the same time-points. On the contralateral side of the dentate gyrus there was an increase in both synapsin I and syntaxin 1B at 5 h only. All of these long-term potentiation-induced changes were prevented when the tetanus was delivered in the presence of the N-methyl-D aspartate receptor antagonist. (D(-)-2-amino-5-phosphonopentanoic acid. Immunocytochemical staining revealed that protein levels for both synapsin I and syntaxin 1B were elevated in the mossy fibre terminal zone of CA3 5 h after the induction of long-term potentiation. In addition to these plasticity-induced changes, a transient increase in the messenger RNA encoding syntaxin 1B was observed at 2 h in conditions of high intensity stimulation of the perforant path to increase the level of cellular activation, but this change was not maintained even when high intensity stimulation was sustained for 5 h. No changes in either of the messenger RNAs were observed under low frequency stimulation and pseudotetanus at either time-points. These results show that an overall increase in neuronal excitation within a neuronal network can be differentiated from a change in synaptic strength at a specific subset of the synapses, where only synaptic plasticity leads to long-term changes in the expression of selective members of the exocytotic machinery. Altered concentrations of key vesicle proteins may thus provide the means for modulation of neurotransmitter release over long time-periods. The persistent long-term potentiation-induced postsynaptic increase in messenger RNAs encoding these presynaptic proteins has important implications for the propagation of signals downstream from the site of long-term potentiation induction in hippocampal neural networks, and highlights a candidate molecular mechanism for mediating the propagation of synaptic plasticity in such networks. PMID- 9200719 TI - Inhibition by compactin demonstrates a requirement of isoprenoid metabolism for long-term potentiation in rat hippocampal slices. AB - Hippocampal long-term potentiation of synaptic transmission is the primary experimental model of learning and memory in the vertebrate brain. However, the detailed intracellular mechanisms giving rise to this persistent increase in synaptic efficacy remain incompletely understood. Mevalonic acid constitutes the basic precursor not only for cholesterol, dolichol and ubichinone but also for farnesyl-pyrophosphate and geranylgeranylpyrophosphate, which are required for post-translational modification of proteins. We have used the specific 3-hydroxy 3-methylglutaryl-CoA reductase inhibitor, compactin, to examine the role of isoprenoid metabolism for long-term potentiation in rat hippocampal slices. Compactin was applied at a concentration of 25 microM for 70 min before and during tetanization and the orthodromic population spike amplitude and field excitatory postsynaptic potentials were recorded from CA1 pyramidal cells. Compactin had no effect on the initial tetanization. However, compactin-treated slices were not able to maintain long-term potentiation for more than 60 min and population spike as well as field excitatory postsynaptic potentiation returned to basal levels after 120 min. When the slices were retetanized after 180 min, an almost full potentiation of the population spike and an only partial potentiation of the field excitatory postsynaptic potentials were observed. These results suggest an essential role of isoprenoid intermediates for maintenance of hippocampal long-term potentiation. PMID- 9200720 TI - Imaging free zinc in synaptic terminals in live hippocampal slices. AB - Some glutamatergic synapses in the mammalian central nervous system exhibit high levels of free ionic zinc in their synaptic vesicles. The precise role of this vesicular zinc remains obscure, despite suggestive evidence for zinc as a neuromodulator. As a step towards elucidating the role of free zinc in the brain we have developed a method for imaging zinc release in live brain slices. A newly synthesized zinc-sensitive fluorescent probe, N-(6-methoxy-8-quinolyl)-p carboxybenzoylsulphonamide (TFLZn), was used to monitor intracellular zinc in live rat hippocampal slices. The dye loaded into the zinc-rich synaptic vesicles of the mossy fibre terminals in the hippocampal formation. Direct electrical stimulation of the mossy fibre pathway diminished the fluorescence in the mossy fibre terminals, consistent with a stimulus-dependent zinc release. The synaptic release of zinc was followed by the rapid replenishment of the zinc levels in vesicles from an as yet unidentified intracellular zinc source. Furthermore, we present evidence that zinc may play a role in a form of long-term potentiation exhibited by the mossy fibre pathway. PMID- 9200721 TI - Differential expression of apolipoprotein D and apolipoprotein E in the kainic acid-lesioned rat hippocampus. AB - Expression of apolipoprotein D, a member of the lipocalin superfamily of transporter proteins, was investigated in the kainic acid-lesioned rat hippocampus. Using an anti-rat apolipoprotein D antibody and biotin avidin enhanced immunocytochemistry, in the normal rat hippocampus there was little apolipoprotein D expression, that was restricted mainly to scattered astrocytes. By contrast, kainic acid-injected rats showed apolipoprotein D immunoreactivity in the pyramidal neurons of the affected CA fields 24-48 h after injection of the excitotoxin, at a time when there was no histological evidence of cell death. Apolipoprotein D immunoreactivity peaked by day 3, coincident with neuronal cell death, and declined thereafter, reaching very low levels by day 7. Besides pyramidal neurons, apolipoprotein D immunoreactivity was also observed in a small number of reactive glial cells in the affected CA fields, but not in the vascular compartments at any time-point. In contrast to the neuronal expression of apolipoprotein D, apolipoprotein E immunoreactivity was observed predominantly in degenerating astrocytes. In conclusion, following excitotoxic injury with kainic acid, apolipoprotein D is expressed in hippocampal pyramidal neurons destined for subsequent cell death. PMID- 9200722 TI - Atypical antipsychotics block the excitatory effects of serotonin in septohippocampal neurons in the rat. AB - We recently reported that serotonin excites a subpopulation of GABAergic neurons in the rat medial septum/diagonal band of Broca complex via multiple serotonin receptors, including the serotonin2A subtype. Since a subpopulation of medial septum/diagonal band GABAergic neurons projects to the hippocampus, in the present study we tested the effect of serotonin on antidromically-activated septohippocampal neurons using extracellular recordings. Bath-applied serotonin had an excitatory effect in a majority of septohippocampal neurons; serotonin excited septohippocampal neurons had a mean conduction velocity -1.63 +/- 0.07 m/s (n=101). Pharmacologically, MDL 100,907, a selective serotonin2A antagonist blocked the excitatory effect of serotonin in 78% of septohippocampal neurons tested, with a mean pA2 of 8.51 +/- 0.12 (n=22). Additionally, the atypical antipsychotics risperidone and clozapine but not the typical antipsychotic haloperidol, blocked the excitatory effects of serotonin at clinically relevant concentrations. The pA2 values of 8.84 +/- 0.11, 6.57 +/- 0.13 and 5.94 +/- 0.27 for risperidone, clozapine and haloperidol, respectively, obtained in the present study, give a rank order of potency risperidone (1.6 nM) clozapine (269 nM) haloperidol (1.1 microM) which corresponds to that reported in binding studies. Additionally, in whole-cell patch-clamp recordings, risperidone (10 nM) blocked serotonin-induced increase in GABAergic synaptic currents. In conclusion, serotonin excites septohippocampal neurons primarily via the serotonin2A receptor and atypical antipsychotics block this excitation at clinically relevant concentrations. PMID- 9200723 TI - Intracerebral grafts promote recovery of the cholinergic innervation of the hippocampal formation in rats withdrawn from chronic alcohol intake. An immunocytochemical study. AB - We have previously found that alcohol withdrawal aggravates the neuronal cell loss induced by chronic alcohol consumption in the rat hippocampal formation. We have also shown that intracerebral grafts of immature hippocampal tissue could reverse the progressive degeneration that occurs during this withdrawal. Furthermore, we have shown that chronic alcohol consumption reduces the areal density of choline acetyltransferase-immunoreactive neurons and the density of choline acetyltransferase-immunoreactive fibres in the hippocampal formation. Thus, we thought it would be of interest to investigate the effects of alcohol withdrawal in the hippocampal cholinergic innervation and to determine whether the intracerebral grafting of immature hippocampal tissue would have beneficial effects upon the cholinergic system in this condition. Choline acetyltransferase immunoreactive fibres and perikarya were analysed in 14-month-old control, alcohol-fed, withdrawal and withdrawal-grafted groups of rats. The areal density of choline acetyltransferase-immunoreactive neurons was reduced in all experimental groups when compared to controls. The density of choline acetyltransferase-immunoreactive fibres was lower in the alcohol-fed and withdrawal groups than in the control and withdrawal-grafted groups. We conclude that the grafted tissue probably produced neurotrophic factors which allowed a recovery of the hippocampal cholinergic fibre network. This recovery might be of importance to reverse the cognitive dysfunction described after chronic alcohol consumption and withdrawal. PMID- 9200724 TI - Developmental regulation and cell-specific expression of N-methyl-D-aspartate receptor splice variants in rat hippocampus. AB - The present study demonstrates cell-specific and developmental regulation of 5' and 3' splicing of the N-methyl-D-aspartate receptor NR1 subunit within specific neuronal populations of the hippocampus. At birth, NR1 transcripts lacking exon 5 (encoding the amino-terminal splice cassette N1) exhibit mature patterns of labelling within the hippocampus, with high levels of expression in the CA1, CA3, and dentate gyrus. In contrast, exon 5-containing (NR1(1XX)) transcripts are expressed at low levels until P8, at which time expression is prominent and essentially uniform in the CA1, CA3, and dentate gyrus. Exon 5 expression increases at a faster rate in CA3 than in CA1 or dentate gyrus. By the third week postnatal (postnatal day P21), exon 5-containing transcripts exhibit a distinct gradient of labelling, with more intense expression in CA3, than in CA1 or dentate gyrus. By P21 pyramidal neurons of the CA1 and granule cells of the dentate gyrus express mainly NR1(0XX) receptor messenger RNAs (lacking exon 5). Because splicing in of the N1 splice cassette confers greater current amplitude and enhanced potentiation by protein kinase C, these observations predict elevated levels of synaptic activity in the CA1 early in postnatal life, a time at which synaptic plasticity is enhanced. The carboxy-terminal splice cassettes C1 and C2 are regulated independently within the hippocampus. Whereas NR1(X11) (C1-, C2-containing) and NR1(X01) (C2 only) receptors exhibit high levels of expression in CA1, CA3, and dentate gyrus, NR1(X00) receptors are expressed more intensely in pyramidal neurons of CA3. NR1(X10) receptor expression is very low in all cells and at all times examined, even in adults. Because splicing in of the C1 cassette is thought to regulate receptor targeting, clustering, and cytoskeletal interactions, N-methyl-D-aspartate receptors in the two hippocampal subfields may play differing roles in synaptogenesis and the formation of new neuronal contacts. Moreover, cell-specific patterns of NR1(X11) receptor messenger RNAs parallel those of NR1(0XX) receptor messenger RNAs; and cell specific patterns of NR1(1XX) (N1-containing) receptor messenger RNAs parallels those of NR1(X00) (C1-, C2-lacking) receptor messenger RNAs throughout development. These observations suggest that NR1(100) receptors, which exhibits the greatest potentiation by protein kinase C, are likely to be important in CA1 during the second and third weeks postnatal. Cell-specific expression of NR1 splice variants undoubtedly contributes to functional diversity of N-methyl-D aspartate receptor properties in neuronal populations within the hippocampus. Developmental regulation of NR1 splicing is likely to influence synaptic plasticity and the formation of new synaptic contacts. Moreover, findings from this study suggest that a change in NR1 splicing following a neurological injury could significantly alter glutamate pathogenicity in a particular population of cells. PMID- 9200725 TI - Expression of non-N-methyl-D-aspartate glutamate receptor subunits in the olfactory epithelium. AB - The channel properties of the multimeric ionotropic glutamate receptors can be regulated by their subunit composition. The relationship between the structure and physiological functions of glutamate receptors, however, is difficult to study in the CNS because of the large number of these subunits, their widespread distribution, and neuronal heterogeneity. To avoid these difficulties, and to uncover possible novel functions of ionotropic glutamate receptors in sensory neurons, we examined the expression of non-N-methyl-D-aspartate glutamate receptor subunits in a simple neuronal system: the olfactory epithelium. It contains only one neuronal type, the olfactory receptor neuron, that receives no synaptic innervation within the epithelium and therefore should not require conventional postsynaptic glutamate receptors. The axons of these neurons, however, terminate and release glutamate in the glomerular region of the olfactory bulb, and may contain presynaptic glutamate receptors. By reverse transcriptase-polymerase chain reaction amplification and RNase protection assays, we showed that a subset of non-N-methyl-D-aspartate receptor subunits is expressed in the olfactory epithelium. The most abundant is KA2, which can form kainate-selective ion channels with GluR5 or GluR6. Messenger RNAs for GluR6, and for the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate/kainate-type (AMPA/KA) GluR2 and GluR3 subunits, are also present, but at levels lower than that of KA2 by an order of magnitude. In situ hybridization and immunocytochemistry localized KA2 to only the olfactory receptor neurons, and not to any other cell type in the olfactory epithelium. Surprisingly, antibodies against KA2 or GluR5/6/7 primarily stained the olfactory neuron dendritic knobs that are specialized for odorant signalling at the sensory epithelial lumenal surface, and the olfactory neuron axon bundles that project to the olfactory bulb. The presence of a limited subset of non-N-methyl-D-aspartate receptor subunits in the olfactory epithelium, and the localization of a kainate-selective receptor to both the axons and specialized dendritic knobs of olfactory receptor neurons, which receive no known synaptic input, suggest that these non-N-methyl-D aspartate receptor subtypes may mediate either novel non-synaptic functions in the olfactory neuron dendrites or presynaptic functions in the olfactory nerve terminals or axons. These data also suggest that the olfactory sensory system, possessing a relatively simple anatomical organization and a limited number of glutamate receptor subunits, may be useful for elucidating facets of the complex relationships between subunit composition and physiological function of ionotropic glutamate receptors. PMID- 9200726 TI - Glomerular synaptic responses to olfactory nerve input in rat olfactory bulb slices. AB - In olfactory bulb slices from young rats, the field potential evoked in the glomerular layer by stimulation in the olfactory nerve layer consisted of two negative components: an early component (N1) which was blocked by bath application of the kainate/amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 10 microM), and a late, prolonged component (N2; duration > or = 350 msec) which was unaffected by CNQX, was enhanced by reduction of Mg2+ in the medium, and was blocked by the N-methyl-D-aspartate receptor antagonist DL-2-amino-5 phosphonovalerate (50 microM). A comparison of the glomerular field potentials before and after knife cuts that isolated the glomerular layer from the deeper layers of the olfactory bulb indicated that both N1 and N2 were produced by currents generated, for the most part, within the glomeruli. A laminar analysis of the field potential profiles evoked by olfactory nerve stimulation in standard medium, or in the presence of CNQX, showed that N1 and N2 reversed polarity in the external plexiform and mitral cell layers, suggesting that both components reflected synaptic responses in the distal, apical dendrites of mitral/tufted cells. Simultaneous field potential recordings in the glomerular layer and intracellular recordings in the mitral cell layer showed that: (i) N1 is associated with a brief, short-latency spiking activity of mitral cells, and (ii) N2 is associated with prolonged mitral cell spiking, since N2 and the late cell firing had similar time-courses, and both were blocked by bath applied DL-2-amino 5-phosphonovalerate. Application of the GABA(A) receptor antagonist bicuculline methiodide (10 microM) to standard medium selectively enhanced N2. The enhanced N2 was significantly reduced by DL-2-amino-5-phosphonovalerate. Strychnine, an antagonist of glycine receptors, had similar effects to those of bicuculline, but only at high concentrations that have been previously shown to block GABA(A) receptors; at low concentrations strychnine had no effect. The effects of all drugs tested were reversible. In the rat olfactory bulb, activation of the olfactory nerve evokes a kainate/AMPA receptor-mediated response in the distal, apical dendrites of mitral/tufted cells, followed by a slow N-methyl-D-aspartate receptor-mediated response which triggers prolonged discharge of mitral cells. GABA(A) receptor-mediated inhibition appears to suppress, preferentially, this N methyl-D-aspartate receptor-mediated component. The presence of prolonged N methyl-D-aspartate receptor-mediated postsynaptic activity at the primary synapses of the olfactory system may play a key role in olfactory processing by facilitating synaptic integration and plasticity. PMID- 9200727 TI - Introduction of the glutamate receptor subunit 1 into motor neurons in vitro and in vivo using a recombinant herpes simplex virus. AB - We developed and characterized a recombinant herpes simplex virus vector and used it to introduce the complementary DNA encoding glutamate receptor subunit 1 flip into postmitotic motor neurons. Infection of purified motor neurons in vitro with this vector resulted in selective, high-level expression of glutamate receptor subunit 1 immunoreactivity in nearly 100% of the neurons. Patch-clamp experiments demonstrated that the protein product of the glutamate receptor subunit 1 flip transgene assembles into functional alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) receptor channels. Herpes simplex virus-glutamate receptor subunit 1 flip was introduced into spinal cord cells by direct injection into the ventral horn and selectively into motor neurons by sciatic nerve injection. High levels of expression were sustained for at least one week and were accompanied by changes in the ionic permeability of AMPA receptors in transgene-expressing neurons. Throughout the first week of infection, there was little evidence for toxicity. Herpes simplex virus provides a versatile tool for manipulating the glutamate receptor phenotype of postmitotic neurons and will permit study of the role of individual glutamate receptor subunits in neuronal physiology and pathophysiology. PMID- 9200728 TI - Membrane properties and inhibitory connections of normal and upper cervically axotomized rubrospinal neurons in the rat. AB - Membrane properties and inhibitory synaptic connections of normal and axotomized rat rubrospinal neurons were examined using a coronal slice preparation. Rubrospinal neurons were axotomized at the C2 vertebral level in vivo. Retrograde labelling in vivo and intracellular biocytin injection following recording were combined to identify recorded axotomized rubrospinal neurons. Their input resistances decreased three and four days and became higher than normal four and 10 weeks following lesioning which coincided with a sequential increase and decrease of their soma area. On the other hand, although their membrane time constant was reduced three and four days following lesioning, it returned to normal value four and 10 weeks following axotomy. Other than these, their membrane current-voltage relationship including an inward rectification in the hyperpolarizing direction was not altered. Normal rubrospinal neurons generated very fast spikes which were not affected by axotomy. Both normal and axotomized cells generated trains of repetitive spikes with a fast spike frequency adaptation at the beginning upon suprathreshold current injection. However, the slope of the steady-state spike frequency and applied current relationship was increased four and 10 weeks following axotomy which also showed an increased steady-state spike frequency in response to high-amplitude current injection. Synaptically, the amplitude and duration of the monosynaptic inhibitory potential evoked from nearby reticular formation were reduced following axotomy. In addition, fewer rubrospinal neurons were found to receive this inhibition 10 weeks following axotomy. Thus, our results show that spinal axotomy induces a time-dependent modification of the membrane properties and spike generating behaviour of rubrospinal neurons which probably represents an initial decrease and a later increase of their excitability. This is accompanied by a persistent decrease of synaptic inhibition which is expected to affect structures that remained innervated by the undamaged axon collaterals of these spinally axotomized neurons. PMID- 9200729 TI - Protein synthesis inhibitors delay transneuronal death in the piriform cortex of young adult rats. AB - It has been demonstrated that apoptotic cell death is an active process that is dependent on RNA and protein synthesis. The question remains as to whether neuronal death in adult, mammalian brains can also be demonstrated in vivo to be dependent on protein synthesis. To address this question we have analysed transneuronal death in the piriform (olfactory) cortex. Following unilateral olfactory bulb ablation in young adult rats, layer IIa of the piriform cortex undergoes rapid degeneration, that commences 12 h after ablation and that is almost complete at 48 h. In order to block protein synthesis, three to six subcutaneous injections of the short acting protein synthesis inhibitor anisomycin, were given at 2 h intervals beginning just before the ablation of the olfactory bulb. In other cases a single injection of the long acting protein synthesis inhibitor emetine were made intracerebrally just before or after olfactory bulb ablation. The number of dying cells was then counted in sections through the rostrocaudal extent of the piriform cortex. Both anisomycin and emetine injections markedly reduced the number of pyknotic cells in layer IIa of the piriform cortex after olfactory bulb ablation. The effect of anisomycin was dose-dependent, near lethal doses leading to an almost complete absence of cell death (six injections of 100 mg/kg). As the doses of anisomycin were reduced, more dying cells were observed. Emetine was only effective at near lethal doses (10 mg/kg) and showed a greater capacity to reduce the levels of cell death when injected into structures near the piriform cortex (e.g., accumbens nucleus) than when injected into more distant structures. To further confirm that the cell death observed was due to apoptosis, we analysed sections by tunel staining to demonstrate DNA fragmentation. We found that tunel-positive cells were also always pyknotic, one of the landmarks of apoptosis. The appearance of pyknotic cells labelled by the tunel method demonstrated that the dying cells in the piriform cortex did indeed undergo apoptosis. PMID- 9200730 TI - Tolerance to diazepam and changes in GABA(A) receptor subunit expression in rat neocortical areas. AB - Long-term treatment with diazepam, a full allosteric modulator of the GABA(A) receptor, results in tolerance to its anticonvulsant effects, whereas an equipotent treatment with the partial allosteric modulator imidazenil does not produce tolerance. Use of subunit-specific antibodies linked to gold particles allowed an immunocytochemical estimation of the expression density of the alpha1, alpha2, alpha3, alpha5, gamma(2L&S) and beta(2/3) subunits of the GABA(A) receptor in the frontoparietal motor and frontoparietal somatosensory cortices of rats that received long-term treatment with vehicle, diazepam (three times daily for 14 days, doses increasing from 17.6 to 70.4 micromol/kg), or imidazenil (three times daily for 14 days, doses increasing from 2.5 to 10.0 micromol/kg). In this study, tolerance to diazepam was associated with a selective decrease (37%) in the expression of the alpha1 subunit in layers III-IV of the frontoparietal motor cortex, and a concomitant increase in the expression of the alpha5 (150%), gamma(2L&S) and beta(2/3) subunits (48%); an increase in alpha5 subunits was measured in all cortical layers. In the frontoparietal somatosensory cortex, diazepam-tolerant rats had a 221% increase in the expression of alpha5 subunits in all cortical layers, as well as a 35% increase in the expression of alpha3 subunits restricted to layers V-VI. Western blot analysis substantiated that these diazepam-induced changes reflected the expression of full subunit molecules. Rats that received equipotent treatment with imidazenil did not become tolerant to its anticonvulsant properties, and did not show significant changes in the expression of any of the GABA(A) receptor subunits studied, with the exception of a small decrease in alpha2 subunits in cortical layers V-VI of the frontoparietal somatosensory cortex. The results of this study suggest that tolerance to benzodiazepines may be associated with select changes in subunit abundance, leading to the expression of different GABA(A) receptor subtypes in specific brain areas. These changes might be mediated by a unique homeostatic mechanism regulating the expression of GABA(A) receptor subtypes that maintain specific functional features of GABAergic function in cortical cell layers. PMID- 9200731 TI - The corticosterone synthesis inhibitor metyrapone decreases dopamine D1 receptors in the rat brain. AB - Experiments were performed to examine the effect of metyrapone, an inhibitor of corticosterone synthesis, on the level of dopamine D1 receptors and their transcripts in the caudate-putamen, nucleus accumbens and olfactory tubercle of the rat brain. The binding to dopamine D1 receptors was measured by receptor autoradiography using the specific D1 receptor antagonist [3H]SCH 23390. The level of dopamine D1 receptor messenger RNA was determined by in situ hybridization histochemistry. The results obtained have shown that metyrapone (two injections of 150 and 50 mg/kg, i.p., given 20 and 3 h before killing, respectively) induced a decrease in the D1 receptor-specific binding in the studied areas of the rat brain. In the caudate putamen, the decrease in [3H]SCH 23390 binding was stronger in the medial (31-39%) than in the lateral part (24 27%). Decreases similar to those in the caudate-putamen were observed in the nucleus accumbens (21%) and olfactory tubercle (32%). Furthermore, metyrapone decreased the level of dopamine D1 receptor messenger RNA in the caudate putamen (17-28%), nucleus accumbens (20%) and olfactory tubercle (18%). In conclusion, our study indicates that glucocorticoids might be involved in the regulation of dopamine D1 receptor level in the rat brain. since metyrapone (which inhibits the synthesis of these hormones) decreases the messenger RNA encoding D1 receptor synthesis, as well as the specific binding to this receptor. PMID- 9200732 TI - Continuous or pulsatile chronic D2 dopamine receptor agonist (U91356A) treatment of drug-naive 4-phenyl-1,2,3,6-tetrahydropyridine monkeys differentially regulates brain D1 and D2 receptor expression: in situ hybridization histochemical analysis. AB - The effect of a chronic D2 dopamine receptor agonist (U91356A) treatment on dopamine receptor gene expression in the brain of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP)-lesioned monkeys was investigated using quantitative in situ hybridization histochemistry. U91356A was administered to MPTP-monkeys for 27 days in a pulsatile (n=3) or continuous (n=3) schedule. Animals treated in a pulsatile mode showed progressive sensitization and developed dyskinesia; whereas with the continuous mode behavioural tolerance was observed but no dyskinesia developed. Untreated MPTP as well as naive control animals were also studied. The efficacy and uniformity of the MPTP effect was assessed by measures of dopamine concentrations by high performance liquid chromatography with electrochemical detection in the relevant brain areas. D1 and D2 receptor messenger RNAs levels were examined by in situ hybridization histochemistry using human complementary RNA probes. Intense specific labelling for D1 and D2 receptor messenger RNAs was measured in the caudate and putamen with a rostrocaudal gradient for D2 receptors and a lower density in the cortex for D1 receptors messenger RNA. D1 receptor mRNA levels in rostral striatum and cortex decreased whereas D2 receptor messenger RNA in caudal striatum increased in MPTP-monkeys compared to control animals. Continuous administration of U91356A reversed the MPTP-induced increase of D2 receptor messenger RNA, whereas the pulsatile administration did not significantly correct these messenger RNA changes. U91356A treatment whether continuous or pulsatile partially corrected the D1 receptor messenger RNA lesion induced decrease in the striatum, whereas no correction was observed in the cortex. All MPTP-monkeys were extensively and similarly denervated suggesting that the D1 and D2 receptor expression changes following U91356A administration were treatment related. Our data show a lesion-induced imbalance of D1 (decrease) and D2 (increase) receptor messenger RNAs in the striatum of MPTP-monkeys. The response of these receptors to D1 agonist treatment showed receptor selectivity and was influenced by the time-course of drug delivery. Hence chronic continuous but not pulsatile administration of U91356A reversed the striatal D1 receptor messenger RNA increase. PMID- 9200733 TI - Brain-derived neurotrophic factor regulates maturation of the DARPP-32 phenotype in striatal medium spiny neurons: studies in vivo and in vitro. AB - The medium spiny neuron is the predominant striatal neuronal subtype. The striatum, a participant in motor and cognitive functions, is a site of pathophysiology in prevalent neuropsychiatric diseases and is the target of many currently utilized pharmacologic agents. DARPP-32, a dopamine and cyclic AMP regulated phosphoprotein, is a widely-used marker of mature striatal medium-sized neurons, but the molecules regulating DARPP-32 transcription have not been identified. We show that a null mutation in the mouse brain-derived neurotrophic factor gene leads to decreased DARPP-32 immunoreactivity in striatal medium spiny neurons at birth and postnatal day 10. Striatal DARPP-32 messenger RNA and protein are decreased relative to wild-type littermate controls. In densely plated (1 x 10(6) cells/cm2) primary cultures derived from the ganglionic eminences, addition of brain-derived neurotrophic factor (100 ng/ml) to defined media results in a greater than 3-fold increase in the number of DARPP-32 immunopositive cells after 12 h and greater than 4-fold (P<0.005) after 24 h. The increase in DARPP-32-immunopositivity is abolished by the addition of 2 microg/ml actinomycin D without a significant effect on cell viability. These data suggest that brain-derived neurotrophic factor directly or indirectly regulates DARPP-32 transcription in medium spiny neurons. This is the first demonstration of transcriptional regulation of DARPP-32, and the first evidence of a forebrain abnormality in a newborn neurotrophin "knockout" mouse. PMID- 9200734 TI - The interleukin-1beta-mediated regulation of proenkephalin and opioid receptor messenger RNA in primary astrocyte-enriched cultures. AB - Opioids have been found to modulate the function of the immune system by regulating the biochemical and proliferative properties of its cellular components. The interaction of opioid and immune systems, however, is not unidirectional, but rather, bidirectional in nature. In the CNS, one cellular target of immune system activation is the astrocytes, glial cells known to synthesize proenkephalin. We have recently shown that these cells also express the messenger RNA transcripts for the opioid receptors mu, delta and kappa, raising the question of the functional significance of this opioid peptide and the related receptors in the astrocytes. That is, why do astrocytes express proenkephalin and opioid receptors, and are these molecules responsive to a factor to which the astrocytes could be exposed in vivo? Furthermore, do these molecules respond to this factor in a region-specific fashion? In the present study, in order to characterize the astrocytic opioid response to an immune factor, we examined the concomitant regulation of mu, delta, kappa and proenkephalin messenger RNAs by interleukin-1beta (1 ng/ml=60 pM, 24 h) in primary astrocyte-enriched cultures derived from the rat (post-natal day 1-2) cortex, striatum, cerebellum, hippocampus and hypothalamus. Interleukin-1beta treatment was found to increase by 55-75% the level of mu receptor messenger RNA in striatal, cerebellar and hippocampal cultures, but not in cultures derived from the cortex or hypothalamus. However, the cytokine had no effect on the level of delta receptor messenger RNA in any of the five cultures examined. In marked contrast to its stimulatory effects on mu receptor messenger RNA levels and its lack of an effect on 6 receptor messenger RNA expression, interleukin-1beta reduced to 10-30% of control levels the kappa receptor messenger RNA levels in all cultures. Interleukin-1beta had no effect on the level of proenkephalin messenger RNA in cortical, striatal, cerebellar and hypothalamic cultures, but did significantly decrease the expression of proenkephalin messenger RNA in hippocampal cultures to 40% of the control level. Therefore, interleukin-1beta differentially regulated opioid receptor messenger RNA in astrocyte-enriched cultures in a manner dependent upon both the receptor type and the brain region from which the culture was derived. The cytokine also differentially regulated proenkephalin messenger RNA in a region-dependent fashion. These findings suggest a capacity for astrocytes to differentially regulate opioid peptide and receptor messenger RNAs in response to an immune factor, supporting the potential existence of a novel immune-opioid system interaction in the CNS. PMID- 9200735 TI - Insulin-like growth factor-I is an osmoprotectant in human neuroblastoma cells. AB - A role in neuronal homeostasis is suggested by the persistent expression of the insulin-like growth factors in the adult nervous system. SH-SY5Y human neuroblastoma cells, a well-characterized in vitro model of human neurons, were used to investigate the effects of hyperosmotic stress on neurons. Neuronal DNA fragmentation was detected within 1 h and pyknotic nuclei were apparent in attached cells after 12 h of hyperosmotic stress. In parallel, flow cytometry measurements revealed a sudden increase in the rate of cells irreversibly undergoing programmed cell death after 12 h of hyperosmotic exposure. Insulin like growth factor-I delayed the onset of a laddered DNA fragmentation pattern for 24 h and provided continuing protection against hyperosmotic exposure for 72 h. Amino acid uptake was decreased in hyperosmotic medium even in the presence of insulin-like growth factor-I; the protein synthesis inhibitor cycloheximide neither prevented the induction of programmed cell death nor interfered with the ability of insulin-like growth factor-I to act as an osmoprotectant in hyperosmotic medium. Cysteine and serine protease inhibitors each prevented DNA fragmentation under hyperosmotic conditions, suggesting that the osmoprotectant activity of insulin-like growth factor-I involves the suppression of protease activity. Collectively, these results indicate that insulin-like growth factor-I limits the death of neurons under stressful environmental conditions, suggesting that it may provide a candidate therapy in the treatment of hyperosmolar coupled neurological injury. PMID- 9200736 TI - Induction of apoptosis in vitro and in vivo by the cholinergic neurotoxin ethylcholine aziridinium. AB - The patterns of cell death induced by the cholinergic neurotoxin ethylcholine aziridinium have been investigated in vitro and in vivo. In vitro, the drug induced apoptosis both in neuronal SK-N-MC cells (human neuroblastoma cells) and in non-neuronal 293 cells (a human embryonic kidney cell line). Apoptosis was developed maximally between 15 and 24 h of exposure to ethylcholine aziridinium (100 microM). At the ultrastructural level apoptotic cells were characterized by condensation and margination of nuclear chromatin, fragmentation of nuclei and the formation of apoptotic bodies. Inhibition of endonuclease by zinc almost completely prevented the occurrence of apoptosis. The free radical scavenger Tempol effectively inhibited ethylcholine aziridinium-induced apoptosis by 78.6 +/- 10.3% (n=4), whereas cycloheximide and actinomycin D were only partially effective. In vivo, following injection of ethylcholine aziridinium (2 nmol) into the lateral ventricle of rat brain a high incidence of apoptotic cells as verified by in situ tailing was visible in the periventricular tissue. Neurons as well as glia were affected by the neurotoxin. The number of apoptotic cells peaked two to three days after injection of ethylcholine aziridinium and declined thereafter. Up to one week after ethylcholine aziridinium no signs for the induction of apoptosis in the medial septal nucleus were found. This study provides clear evidence that a neurotoxic compound that induces programmed cell death in vitro is likely to have the same capacity in vivo. Yet, in the case of ethylcholine aziridinium, both the in vitro and the in vivo induction of programmed cell death appears to be an additional feature of ethylcholine aziridinium, which may be independent of the well-established degenerative effect of ethylcholine aziridinium on the cholinergic septohippocampal pathway. The present data indicate that ethylcholine aziridinium provides a useful tool to study molecular mechanisms of neuronal apoptosis. PMID- 9200737 TI - Angiotensin II interacts with nitric oxide-cyclic GMP pathway in the central control of drinking behaviour: mapping with c-fos and NADPH-diaphorase. AB - Recognition of the role of nitric oxide in cell-to-cell communication has changed the concept of traditional neurotransmission. We have shown previously that N methyl-D-aspartate receptors mediate dipsogenic responses and c-Fos expression induced by intracerebroventricular infusion of angiotensin II. Since these receptors are known to be linked to the nitric oxide-cyclic GMP pathway, the present study explores the contribution of this path to the behavioural and cellular effects of intracerebroventricular angiotensin II by using behavioural testing, NADPH-diaphorase histochemistry and immunocytochemical staining for the immediate-early gene, c-fos. N(G)-nitro-L-arginine methyl ester (125 and 250 microg, intracerebroventricular), an inhibitor of nitric oxide synthase, and Methylene Blue (100 microg), an inhibitor of guanylate cyclase activation, antagonized water intake induced by intracerebroventricular injection of 25 pmol angiotensin II. The effects of N(G)-nitro-L-arginine methyl ester were reversed by co-injection of L-arginine, the substrate for nitric oxide synthase. However, N(G)-nitro-L-arginine methyl ester did not alter the pattern of angiotensin II induced c-fos expression in the organum vasculosum of the lamina terminalis, median preoptic nucleus, hypothalamic paraventricular nucleus and supraoptic nucleus. Double staining with NADPH-diaphorase histochemistry and c-Fos immunocytochemistry showed that neurons staining for both were localized to the anterior third ventricle. However, only 19-25% of the c-Fos-positive neurons expressed NADPH. There were also substantial numbers of neurons in which angiotensin II induced c-Fos that were NADPH-negative. Extensive co-distribution of NADPH-diaphorase-stained cells and those expressing c-fos in response to intracerebroventricular injection of angiotensin II, especially in the median preoptic nucleus, imply that nitric oxide might participate in the mechanism of angiotensin II-induced drinking behaviour. However, a low rate of co-localization of the two markers to individual cells suggests that angiotensin II stimulated the production of nitric oxide and c-Fos in different populations of neurons. Since our previous results showed that glutamate blockade, but not nitric oxide synthase inhibition, suppressed angiotensin II-induced c-Fos, the experiments reported here further suggest that nitric oxide release is not an essential requirement for the expression of c-fos elicited by angiotensin II. They also provide evidence that the dipsogenic and c-Fos responses to angiotensin II are dissociated at a cellular level. PMID- 9200738 TI - Long-lasting effect of catecholamine deficiency on differentiating vasopressin and oxytocin neurons in the rat supraoptic nucleus. AB - According to our earlier study, the catecholamine depletion in neonatal rats resulted in stimulation of the vasopressin and oxytocin gene expression in the neurons of the supraoptic nucleus. The present study extends this line, evaluating whether the catecholamine deficiency provides a long-lasting effect on the differentiating vasopressin and oxytocin neurons of the supraoptic nucleus. Catecholamines were depleted by daily injections of an inhibitor of the catecholamine synthesis, alpha-methyl-p-tyrosine, first, to pregnant rats from the 9th to the 21st day of gestation and, then, to their pups from the 2nd to the 10th postnatal day. The animals, injected with saline instead of drugs, served as controls. The pharmacologically-treated and control rats were kept for four months under normal laboratory conditions until processing the materials for semi quantitative in situ hybridization and immunocytochemistry of vasopressin and oxytocin messenger RNAs and peptides, respectively. There were no differences in the vasopressin and oxytocin messenger RNA concentrations in the supraoptic nucleus in rats following preliminary catecholamine depletion compared to controls. Conversely, the catecholamine deficiency resulted in an increased content of the vasopressin-immunoreactive material in cell bodies and processes. This was also the case for the oxytocin-immunoreactive cell bodies but only in females, suggesting an interference of catecholamines with sexual steroids in their action. The number and size of vasopressin and oxytocin neurons did not change in pharmacologically-treated rats compared to the controls. Thus, the catecholamine deficiency in the course of the neuron differentiation resulted in a long-lasting augmentation of the intracellular content of vasopressin and oxytocin but did not influence the vasopressin and oxytocin gene expression. This might be explained rather by the reduced level of peptide release than by an increased level of the peptide production. PMID- 9200739 TI - Serotonergic regulation of circadian rhythms in Syrian hamsters. AB - This study investigated the effects of (+/-)-2-dipropylamino-8-hydroxy-1,2,3,4 tetrahydronaphthaline hydrobromide (8-OH-DPAT) on circadian rhythms in Syrian hamsters. Systemic administration of 8-OH-DPAT (0.75 mg in 150 microl saline) at circadian time 7 produced phase advances in the circadian activity rhythm. These 8-OH-DPAT-induced phase advances were blocked by microinjection of bicuculline (166 ng, 200 nl) into the suprachiasmatic nucleus, suggesting that GABAergic activity in the suprachiasmatic nucleus mediates the phase shifts produced by systemic injections of 8-OH-DPAT. Microinjection of 8-OH-DPAT (1 microg, 200 nl) or serotonin (0.7 microg, 200 nl) directly into the suprachiasmatic nucleus did not induce phase shifts at circadian time 7, suggesting that the phase shifting effects of systemic injection of 8-OH-DPAT are mediated outside the suprachiasmatic nucleus. To examine possible sites of action of 8-OH-DPAT, 8-OH DPAT (0.5 microg (100 nl) or 1.0 microg (200 nl)) was microinjected into the intergeniculate leaflet, dorsal raphe nuclei, and the median raphe nucleus at circadian time 7. Significant phase advances were observed after microinjection into the dorsal raphe and median raphe but not the intergeniculate leaflet. These results support the hypothesis that systemic injection of serotonergic agonists can alter circadian rhythms via action in the midbrain raphe nucleus, and that the phase shifts induced by microinjection of 8-OH-DPAT into the raphe nuclei are mediated by a neurotransmitter other than serotonin within the suprachiasmatic nucleus. PMID- 9200740 TI - Basal forebrain and cerebral cortical muscarinic receptors mediate increase in cortical blood flow provoked by periaqueductal gray matter. AB - The midbrain periaqueductal gray matter has been identified as a reflex centre located uppermost in the central organization of diverse defensive reactions. We recently found that when activated, the caudal third of the lateral periaqueductal gray was also capable of provoking a marked increase in cortical blood flow. The response may be the combined outcome of a flow increase of nitrergic origin and that coupled to a possible concomitant cortical activation. In the present study, we attempted to clarify the neural substrates for mediation of the increase in flow (observed by laser-Doppler flowmetry), in 49 anaesthetized, artificially ventilated, and cervically cordotomized rats. The flow increase provoked by stimulation of the particular subdivision of the periaqueductal gray with N-methyl-D-aspartate (1 mM, 100 nl) was unaffected by i.v. pentolinium tartrate (10 mg/kg), suggesting little contribution by the cerebrovasodilator parasympathetic nervous system to the response. The response was abolished by i.v. or topical cortical administration of scopolamine hydrobromide (3.16 mg/kg or 1.0 mM, respectively). Placement of bilateral lesions in the basal forebrain with alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (15 mM) impaired the cortical choline acetyltransferase activity and attenuated the flow response. Overall, we suggest that the cholinergic corticopetal neurons of the nucleus basalis of Meynert and cortical muscarinic receptors may form a principal efferent arm of a central circuitry emanating from the subdivision of the periaqueductal gray, in the mediation of the increase in cortical blood flow and possible cortical activation. PMID- 9200741 TI - Aortic barodenervation up-regulates alpha2-adrenoceptors in the nucleus tractus solitarius and rostral ventrolateral medulla: an autoradiographic study. AB - Earlier findings have shown that alpha2-adrenoceptors in the nucleus tractus solitarius and rostral ventrolateral medulla modulate baroreflexes. The present study investigated whether attenuation of baroreflexes induced by surgical interruption of aortic baroafferents is related to an alteration of alpha2 adrenoceptor binding in these regions of the brainstem. In vitro autoradiography was utilized to assess the density and binding dissociation constant (affinity) of alpha2-adrenoceptors in the rostral ventrolateral medulla and in the middle and rostral portions of the nucleus tractus solitarius of aortic-barodenervated and sham-operated rats. Compared to sham operation, aortic barodenervation caused an acute rise in mean arterial pressure and heart rate and a significant reduction in baroreflex sensitivity. Two days later, mean arterial pressure and heart rate of conscious aortic-barodenervated rats subsided to sham-operated levels, whereas the baroreflex sensitivity remained significantly (P<0.05) reduced when measured by phenylephrine (0.55+/-0.08 vs 1.26+/-0.07 ms/mmHg) or nitroprusside (0.43+/-0.06 vs 1.01+/-0.09ms/mmHg). Examination of brainstem coronal sections obtained from separate groups of rats 48 h after surgery and preincubated with [3H]rauwolscine (0.5-16 nM) revealed that labeling of alpha2 binding sites was saturable and of high affinity. Scatchard analysis of the saturation isotherms obtained from the three brain areas of sham-operated rats showed an uneven distribution of alpha2 binding sites; the rostral nucleus tractus solitarius exhibited the highest density and lowest affinity. Aortic barodenervation caused region-dependent changes in the binding activity of alpha2 adrenoceptors. These changes comprised significant (P<0.05) increases in the density of alpha2-adrenoceptors in the middle nucleus tractus solitarius (436+/ 60 vs 240+/-50 fmol/mg protein) and rostral ventrolateral medulla (350+/-67 vs 194+/-35 fmol/mg protein) compared with sham-operated rats; no significant changes occurred in the rostral nucleus tractus solitarius. The affinity of alpha2 binding sites was not changed by aortic barodenervation in any of the three brain regions. These findings suggest that attenuation of baroreflexes produced by aortic barodenervation coincides with up-regulation of alpha2 adrenoceptors in brainstem areas that play critical roles in the control of cardiovascular functions. PMID- 9200742 TI - Expression and localization of Na+/H+ exchangers in rat central nervous system. AB - Neurons in the central nervous system regulate their intracellular pH using particular membrane proteins of which two, namely the Na+-dependent Cl-/HCO3- exchanger and the Na+/H+ exchanger, are essential. In this study we examined messenger RNA expression and distribution of Na+/H+ exchanger in the newborn rat central nervous system and with maturation using Northern blot analysis and in situ hybridization. Our study clearly shows that each Na+/H+ exchanger has a different expression pattern in the rat central nervous system. As in non excitable tissues, Na+/H+ exchanger 1 is by far the most abundant of all Na+/H+ exchangers in the rat central nervous system. Its expression is ubiquitous although its messenger RNA appears at higher levels in the hippocampus, in the 2nd/3rd layers of periamygdaloid cortex and in the cerebellum. The low level of messenger RNAs encoding Na+/H+ exchanger 2 and 4 is mainly expressed in the cerebral cortex and in the brainstem-diencephalon, while Na+/H+ exchanger 3 transcripts are found only in the cerebellar Purkinje cells. From a developmental point of view, Na+/H+ exchanger 1, 2 and 4 showed an increased level in their transcripts in the cerebral cortex while an opposite trend existed in the cerebellum from postnatal day 0 to postnatal day 30. The messenger RNA for Na+/H+ exchanger 3, however, increased its level with age in cerebellum. From our data we conclude that: i) the expression of the Na+/H+ exchanger is age-, region-, and subtype-specific, with Na+/H+ exchanger 1 being the most prevalent in the rat central nervous system; ii) specialization of groups of neurons with respect to the type of Na+/H+ exchanger is clearly illustrated by Na+/H+ exchanger 3 which is almost totally localized in cerebellar Purkinje cells; and iii) the developmental increase in the messenger RNA for Na+/H+ exchanger 1 in the cerebral cortex and hippocampus is consistent with our previous studies on intracellular pH physiology in neonatal and mature neurons. Together this study indicates that expression of each Na+/H+ exchanger messenger RNA is differentially regulated both during development and in the different regions of rat central nervous system. PMID- 9200743 TI - Trigeminal ganglion innervation of the cochlea--a retrograde transport study. AB - Innervation patterns of sensory nerves from the trigeminal ganglion to the cochlear blood vessels were studied using retrograde transport of wheat germ agglutinin conjugated to horseradish peroxidase. Guinea-pigs (n=7) were unilaterally implanted with an osmotic pump and a cannula for cochlear delivery of 2% or 20% wheat germ agglutinin horseradish peroxidase (Group 1), 2% wheat germ agglutinin-horseradish peroxidase followed by 100 micromol capsaicin (Group 2), or vehicle alone. Histological sections of the trigeminal ganglia, the C1 and C2 dorsal ganglia, the superior and inferior ganglia of the glossopharyngeal nerve bilaterally, the midbrain and the brainstem were obtained after 48 h of infusion. In Group 1, a large number of labeled nerve cell bodies were observed in the anteromedial portion of the trigeminal ganglion and at the origin of the ophthalmic nerve. Some labeled cells were also found on the lateral side of the ophthalmic nerve, as well as on the medial side of the maxillary nerve root. Capsaicin pretreatment significantly reduced the density of labeled neurons in the trigeminal ganglion. A few labeled neurons were also found in the trigeminal brainstem nucleus complex and in certain auditory brainstem nuclei. No wheat germ agglutinin horseradish peroxidase-positive cells were observed in the spinal C1 or C2 cervical ganglia or in the superior or inferior glossopharyngeal ganglia. In contrast, wheat germ agglutinin-horseradish peroxidase application to the middle ear resulted in labeled cells in the middle posterolateral portion of the trigeminal ganglia and in the superior ganglia of the glossopharyngeal nerve. These results provide the first direct evidence that the trigeminal ganglion sends projections to the cochlea. PMID- 9200744 TI - Age-related muscle stiffness: predominance of non-reflex factors. AB - This study was aimed at assessing the contribution of reflex and non-reflex factors to the muscle tone of old female Wistar rats. The hind foot of a rat was flexed or extended at the ankle joint by 25 degrees over 250 ms. The resistance of the foot to passive movements (torque, mechanomyogram), as well as the reflex electromyographic activity in the gastrocnemius and tibialis anterior muscles, were recorded simultaneously. Moreover, the impact of the blockade of the reflex activity caused by the local anesthetic lignocaine (1-2 ml of a 2% solution, injected in the vicinity of the sciatic nerve) on the muscle tone was investigated. Additionally, old rats' hind leg muscle samples were analysed using fluorescent microscopy for the expression of fibronectin, which is an early marker of connective tissue formation. It has been shown that old rats are characterized by (i) a substantially increased resistance of flexor muscle stiffness (measured during extension) and unchanged resistance of extensors (measured during flexion), (ii) the loss of a major part of the reflex electromyographic activity and (iii) the increased content of fibronectin in muscles. Moreover, it has been shown that lignocaine, which completely blocked the electromyographic reflex activity in the gastrocnemius and tibialis anterior muscles in young animals, was unable to counteract the resistance of these muscles to passive movements in old rats. The present results suggest that the muscle stiffness seen in old rats is not due to a reflex response, but depends mainly on non-reflex factors--chiefly on a large overgrowth of non-elastic connective tissue replacing degenerated active muscle fibers. PMID- 9200745 TI - Unique properties of [3H]MK-801 binding in membranes from the rat spinal cord. AB - In order to investigate possible differences between NMDA receptor-coupled ion channels in the spinal cord and in the cerebral cortex, we have characterized [3H]MK-801 [(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine] binding and its regulation by glutamate and glycine in membrane preparations of the rat spinal cord and cerebral cortex. The K(D) value of [3H]MK-801 binding was higher in the spinal cord than in the cerebral cortex, mainly due to a lower association rate constant. When corrected for the concentrations of residual endogenous amino acids, the EC50 values for glycine were lower at spinal NMDA receptors compared to those in the cerebral cortex, whereas the EC50 values for glutamate were similar in both regions. The IC50 values of D-((3)-2 carboxypiperazin-4-yl)-propyl-1-phosphonic acid (D-CPP) were significantly lower in the spinal cord in the presence of saturating concentrations of glutamate. The IC50 values of 7-chloro-4-hydroxy-3-(3-phenoxy)phenyl-2(H)-quinoline (L-701,324) were significantly lower in the spinal cord under all conditions. These results suggest that NMDA receptors in the spinal cord display low affinity for MK-801, which may correspond to a lower affinity of the voltage-dependent Mg2+ block. Furthermore, NMDA receptors in the spinal cord appear to display high sensitivity to glycine and to glutamate and glycine antagonists. PMID- 9200747 TI - Cuneiform nucleus stimulation-induced sympathoexcitation: role of adrenoceptors, excitatory amino acid and serotonin receptors in rat spinal cord. AB - Stimulation of the midbrain cuneiform nucleus has previously been shown to produce increases in arterial blood pressure and lumbar sympathetic nerve activity. While this sympathoexcitatory effect is, in part, due to excitation of premotor sympathoexcitatory neurons in the rostral ventrolateral medulla, the specific spinal neurotransmitter systems recruited by cuneiform nucleus stimulation remains to be elucidated. In this study, mean arterial pressure, resting and cuneiform nucleus stimulation-evoked lumbar sympathetic nerve activity were analysed following intrathecal injections of an excitatory amino acid antagonist (kynurenic acid), alpha1-adrenoceptor antagonist (prazosin) and a serotonin receptor antagonist (methiothepin) in anesthetized, paralysed male Sprague-Dawley rats. Mean arterial pressure and resting sympathetic nerve discharge were decreased by all treatments (n = 6/group) compared to the vehicle control group. Intermittent electrical stimulation of the cuneiform nucleus produced a bimodal sympathoexcitatory response, of which the short latency peak was significantly attenuated (43% reduction) by intrathecal kynurenate whereas the long latency peak was reduced by intrathecal prazosin (decrease of 21%) and methiothepin (38% attenuation). These results are consistent with the significant roles of excitatory amino acid, alpha1-adrenergic and serotonin receptors in modulating the activity of sympathetic vasomotor preganglionic neurons supplying the lumbar sympathetic nerve trunk, and suggest the existence of at least three neuronal groups and/or pathways associated with the sympathoexcitatory response to cuneiform nucleus stimulation. PMID- 9200746 TI - Ultra-low doses of naltrexone or etorphine increase morphine's antinociceptive potency and attenuate tolerance/dependence in mice. AB - In previous studies we showed that low (pM) concentrations of naloxone (NLX), naltrexone (NTX) or etorphine selectively antagonize excitatory, but not inhibitory, opioid receptor-mediated functions in nociceptive types of sensory neurons in culture. Cotreatment of these neurons with pM NTX or etorphine not only results in marked enhancement of the inhibitory potency of acutely applied nM morphine [or other bimodally-acting (inhibitory/excitatory) opioid agonists], but also prevents development of cellular manifestations of tolerance and dependence during chronic exposure to microM morphine. These in vitro studies were confirmed in vivo by demonstrating that acute cotreatment of mice with morphine plus a remarkably low dose of NTX (ca. 10 ng/kg) does, in fact, enhance the antinociceptive potency of morphine, as measured by hot-water tail-flick assays. Furthermore, chronic cotreatment of mice with morphine plus low doses of NTX markedly attenuates development of naloxone-precipitated withdrawal-jumping in physical dependence assays. The present study provides systematic dose response analyses indicating that NTX elicited optimal enhancement of morphine's antinociceptive potency in mice when co-administered (i.p.) at about 100 ng/kg together with morphine (3 mg/kg). Doses of NTX as low as 1 ng/kg or as high as 1 microg/kg were still effective, but to a lesser degree. Oral administration of NTX in the drinking water of mice was equally effective as i.p. injections in enhancing the antinociceptive potency of acute morphine injections and even more effective in attenuating development of tolerance and NLX-precipitated withdrawal jumping during chronic cotreatment. Cotreatment with a subanalgesic dose of etorphine (10 ng/kg) was equally effective as NTX in enhancing morphine's antinociceptive potency and attenuating withdrawal-jumping after chronic exposure. These studies provide a rationale for the clinical use of ultra-low dose NTX or etorphine so as to increase the antinociceptive potency while attenuating the tolerance/dependence liability of morphine or other conventional bimodally-acting opioid analgesics. PMID- 9200748 TI - NH3- and CO2-induced suppression of taste nerve responses in clawed toads and eels. AB - We investigated the effects of intracellular pH values (pHi) on taste nerve responses of clawed toads and eels. (1) CO2, NH3 or trimethylamine reversibly suppressed the taste nerve responses of clawed toads to various amino acids, CaCl2 and caffeine. IC50 values of the suppression of the responses to 0.1 mM L proline were as follows: approximately 10 mM for CO2, approximately 0.3 mM for NH3, approximately 0.2 mM for trimethylamine. (2) Cross-adaptation experiments showed that L-proline, caffeine and CaCl2 stimulated different receptor sites from each other, indicating the suppressive effect was non-specific. (3) Although CO2, NH3 or trimethylamine yielded the charged molecules, HCO3-, CO3(2-), NH4+ or trimethylammonium on hydration, none of these charged species suppressed taste responses. (4) NH3 increased the threshold concentration of L-proline by e-fold per 0.37 mM NH3 and decreased the maximum response to L-proline with increasing NH3 concentration. (5) The taste nerve responses of eels to 0.1 mM L-arginine, a potent stimulus on eel taste receptors, were similarly suppressed by NH3 with an IC50 value of approximately 0.3 mM. (6) These results indicated that these uncharged species changed pHi, which suppressed the taste responses. CO2-induced acidosis and NH3- or trimethylamine-induced alkalosis are likely to inhibit activities of ion channels or enzymes involved in taste transduction mechanisms. PMID- 9200749 TI - Co-localization of vasoactive intestinal polypeptide, gamma-aminobutyric acid and choline acetyltransferase in neocortical interneurons of the adult rat. AB - Interneurons immunoreactive for vasoactive intestinal polypeptide (VIP) are integral elements of columnar organization patterns in the rat cerebral cortex. By application of the sensitive mirror technique, the co-localization of VIP with the classical inhibitory neurotransmitter gamma-aminobutyric acid (GABA) and the acetylcholine-synthesizing enzyme, choline acetyltransferase (ChAT), was investigated in neocortical neurons. Furthermore, the frequency of co localization of ChAT with GABA was determined. In a sample of 118 VIP immunoreactive neurons, mostly from the primary somatosensory cortex, it was demonstrated that virtually all of them reveal immunoreactivity for GABA and, therefore, are to be GABAergic. Moreover, 34% of mostly bipolar, VIP-positive neurons contained ChAT and are, thus, supposedly cholinergic as well. Co localization of VIP and ChAT varied according to cortical laminae. Finally, 88% of a total of 60 ChAT-immunoreactive neurons were also immunostained for GABA. It is concluded that almost all VIP-immunoreactive neurons and most of the cholinergic neurons in rat neocortex represent partly overlapping subpopulations of inhibitory interneurons utilizing GABA. PMID- 9200751 TI - The role of the frontal cortex in the mouse in behavioral sensitization to amphetamine. AB - Pharmacological studies have shown that a variety of neuroeffectors are involved in behavioral sensitization to amphetamine-induced stereotypy. In the present work, the effect of some of these drugs on sensitization was studied after intracortical administration in order to determine the role of the cortex in mediating their systemic effects. The dopamine antagonists sulpiride and spiperone were both ineffective against the acute response to amphetamine; nevertheless, both blocked the induction of sensitization, suggesting that the mesocortical dopamine pathway is not involved in the acute response but is necessary for the induction of sensitization. Both CPP, an NMDA receptor antagonist, and THIP, a GABA(A) agonist, blocked the acute response and the induction of sensitization to amphetamine. On the other hand, mecamylamine, the nicotinic cholinergic antagonist, failed to affect either the acute response or the induction of sensitization, which suggests that the cortex is not a locus of its activity. Anisomycin, an inhibitor of protein synthesis, and diltiazem, a calcium-channel blocker, were both ineffective against the acute response, but both blocked induction. All of the drugs, except CPP and THIP, were ineffective against the expression of sensitization; therefore, the ability of the other drugs to block expression must reside within another locus. Bicuculline injected intracortically in non-convulsant doses produced a stereotypy indistinguishable from that induced by amphetamine; and the effect was readily antagonized by CPP administered either systemically or intracortically. In contrast, sulpiride by either route of administration failed to block the bicuculline-induced stereotypy; we conclude, therefore, that the stereotypic effect of bicuculline is not mediated by dopamine. These results imply that amphetamine-induced stereotypy is mediated in the cortex by the removal of the inhibitory control of the excitatory system. The data also suggest that cortical dopamine, as well as the NMDA and GABA(A) systems, is important in sensitization to amphetamine. In general the data demonstrate that different neuroeffectors involved in sensitization exert their effects at different brain loci. PMID- 9200750 TI - Localization of angiotensin-converting enzyme, angiotensin II, angiotensin II receptor subtypes, and vasopressin in the mouse hypothalamus. AB - The hypothalamic angiotensin II (Ang II) system plays an important role in pituitary hormone release. Little is known about this system in the mouse brain. We studied the distribution of angiotensin-converting-enzyme (ACE), Ang II, Ang II receptor subtypes, and vasopressin in the hypothalamus of adult male mice. Autoradiography of binding of the ACE inhibitor [125I]351A revealed low levels of ACE throughout the hypothalamus. Ang II- and vasopressin-immunoreactive neurons and fibers were detected in the paraventricular, accessory magnocellulary, and supraoptic nuclei, in the retrochiasmatic part of the supraoptic nucleus and in the median eminence. Autoradiography of Ang II receptors was performed using [125I]Sar1-Ang II binding. Ang II receptors were present in the paraventricular, suprachiasmatic, arcuate and dorsomedial nuclei, and in the median eminence. In all areas [125I]Sar1-Ang II binding was displaced by the AT1 receptor antagonist losartan, indicating the presence of AT1 receptors. In the paraventricular nucleus [125I]Sar1-Ang II binding was displaced by Ang II (Ki = 7.6 X 10(-9)) and losartan (Ki = 1.4 X 10(-7)) but also by the AT2 receptor ligand PD 123319 (Ki = 5.0 X 10(-7)). In addition, a low amount of AT2 receptor binding was detected in the paraventricular nucleus using [125I]CGP42112 as radioligand, and the binding was displaced by Ang II (Ki = 2.4 X 10(-9)), CGP42112 (Ki = 7.9 x 10(-10)), and PD123319 (Ki = 2.2 x 10(-7)). ACE, Ang II, and AT1 as well as AT2 receptor subtypes are present in the mouse hypothalamus. Our data are the basis for further studies on the mouse brain Ang II system. PMID- 9200752 TI - The promoting effects of bFGF and astrocyte extracellular matrix on process outgrowth by adult human oligodendrocytes are mediated by protein kinase C. AB - Process extension by oligodendrocytes (OLs) is a critical early step in myelin formation. We have previously reported that the basal- or phorbol ester-enhanced process outgrowth by adult human OLs is mediated by oligodendroglial protein kinase C (PKC). Recently, we demonstrated that astrocytes facilitated process outgrowth by adult human OLs through the interaction between astrocyte-derived basic fibroblast growth factor (bFGF) and astrocyte extracellular matrix (ECM). If PKC is central to the signal transduction cascade that leads to process formation by OLs, then the effects of bFGF and astrocyte ECM should also involve PKC. In the current study, we have addressed the involvement of PKC in the bFGF- and astrocyte ECM- enhanced process formation by adult human OLs by using a selective inhibitor of PKC, calphostin C. The results show that calphostin C dose dependently reduced process extension elicited by bFGF and astrocyte ECM, at IC50 concentrations of 24.5 and 26.6 nM, respectively. At the concentrations of calphostin C that inhibited process extension by adult human OLs, necrosis (measured by lactate dehydrogenase release) and apoptosis (determined by using a fluorescent terminal deoxynucleotidyl transferase assay) of OLs did not occur. Finally, we demonstrate that another specific inhibitor of PKC, CGP 41251, also reduced process formation that is elicited by bFGF and astrocyte ECM. Thus, all process-extending agents for adult human OLs identified to date signal through PKC, further implicating PKC of OLs as being central to the production of process extension, an early event in myelinogenesis. PMID- 9200753 TI - Regulation of cytoskeletal proteins by thyroid hormone during neuronal maturation and differentiation. AB - Primary cultures of neurons from 16- to 17-day-old embryonic rat cerebra were maintained for 3 weeks in thyroid hormone deficient (THdef) and thyroid hormone supplemented (THsup) media to investigate how TH regulates the cytoskeletal (CSK) proteins during neuronal differentiation and maturation. Two distinct phases of regulation of triton-insoluble CSK-proteins by TH were discernible--an early phase (days 4-8 of culture) when TH-deficiency resulted in down-regulation of these proteins and a late phase (days 16-20) involving up-regulation of these proteins. In contrast, the triton-soluble non-CSK proteins always remained up regulated by TH. The two main effects of TH-deficiency were retarded neurite outgrowth and altered neuronal morphology. Of all the CSK proteins, actin was found to be predominantly sensitive. Alterations in the level of CSK actin during neuronal maturation were found to be parallel to changes in steady-state level of actin mRNA as well as actin synthesis. However, these TH-induced changes (up regulation of actin during the early phase and down-regulation during the late phase) did not lead to parallel changes in the level of soluble G-actin which was comparable at both days 8 and 16 in THdef and THsup cultures. Quantitation of different forms of intracellular actin revealed that G-actin level declined by about 50% between days 8 and 16. In the case of THsup neurons, this reduction in G-actin was accompanied by a parallel increase in the non-CSK F-actin, whereas TH deficiency resulted in a corresponding increase in CSK F-actin during the terminal differentiation of neurons. Thus TH regulates the biogenesis of CSK proteins with a predominant effect on actin and the transformation of G-actin into non-CSK F-actin appears to be the key step in neuronal maturation which is affected by hypothyroidism. PMID- 9200754 TI - Changes of forearm EMG and cerebral evoked potentials following sudden muscle stretch during isometric contractions in patients with Parkinson's disease. AB - Various investigators have reported that the late reflex EMG activity following muscle stretch is increased in patients with Parkinson's disease. To explore the basis of this increased activity, we have now recorded the late EMG responses together with associated cerebral responses following muscle stretch in parkinsonian patients. Nine patients and eight controls participated in two sets of experiments in which they grasped a handle attached to a torque motor and maintained the wrist isometrically against a constant flexor force. The force was changed unpredictably (first set) or predictably (second set of experiments), causing a stretch of wrist extensors or flexors. Cerebral responses and muscle responses from the forearm were recorded and averaged separately depending upon condition. The late muscle responses to unpredictable muscle stretch were enhanced in parkinsonian patients while the cerebral responses were attenuated when compared to controls. The alteration of the electrocerebral response began approx. 25 ms prior to the late M2 muscle response. Both controls and patients showed a markedly attenuated cerebral response when the muscle stretch was predictable. These results indicate that the electrocerebral response to muscle stretch is altered prior to the onset of M2 in patients with Parkinson's disease, and suggest that these cerebral events reflect components of a long-latency transcerebral reflex pathway that is affected in this disorder. PMID- 9200755 TI - Selegiline treatment after transient global ischemia in gerbils enhances the survival of CA1 pyramidal cells in the hippocampus. AB - Selegiline (L-deprenyl) has shown neuroprotective effects in a variety of degenerative processes. The present experiments were designed to test whether post-ischemia administered selegiline would alleviate delayed neuronal death of the gerbil hippocampal pyramidal cells following transient global ischemia. Common carotid arteries were occluded for 5 min. Saline or selegiline, 0.25 mg/kg s.c., was administered 2 h after the ischemia followed by a daily injection for either 3 or 7 days. After decapitation, the delayed death of the hippocampal CA1 pyramidal cells was assessed using Nissl-stained sections. In situ hybridization was used to reveal the expression of hsp70 mRNA 1, 3 or 7 days after the ischemia. Animals treated with selegiline for 7 days showed significantly lower damage score (scale 0-3: 0, normal; 1, < 10% of the neurons damaged; 2, 10-50% damaged; 3, > 50% damaged) compared to the saline-treated animals 1.73 +/- 0.18 and 2.41 +/- 0.16 (mean +/- S.E.M., P = 0.0133), respectively. A similar trend was found in animals after the 3-day treatment: 1.68 +/- 0.32 vs. 2.06 +/- 0.25 (P > 0.5). The expression of hsp70 mRNA in the CA1 pyramidal cell layer was strong still 3 days after the ischemic insult but vanished by 7 days. Densitometric measurements using 14C-plastic standards showed that the intensity of the CA1a hsp70 signal on the 3rd day correlated negatively to the cell-damage score (r = -0.72, P < 0.001), suggesting that hsp70 does not serve as a quantitative marker for CA1 neuronal injury in this model. Instead, the hsp70 expression was associated with improved neuronal survival lasting often longer in selegiline-treated animals (P > 0.5). The results show that a low dose of selegiline can alleviate the delayed hippocampal neuronal death in gerbils when administered 2 h after an ischemic insult. PMID- 9200757 TI - Affinity of naloxone and its quaternary analogue for avian central delta and mu opioid receptors. AB - Quaternary narcotic antagonists that are assumed not to penetrate the blood-brain barrier following systemic administration are commonly used to distinguish between peripheral and central actions of opiates. In mammals, these antagonists have a lower affinity for opioid receptors than their tertiary parent compounds. The relative affinity of quaternary vs. tertiary antagonists either for opioid receptors in non-mammalian species or for specific receptor subtypes has, however, not been determined. Using brain tissues from a passerine songbird (Junco hyemalis), we found the affinity of the quaternary antagonist, naloxone methiodide (Nal MI), for brain opioid receptors to be less than 10% that of Nal HCl. Further, Nal MI affinity for mu and delta receptors is 8.7% and 3.7%, respectively, that of Nal HCl. These results confirm that tertiary narcotic antagonist quaternization substantially reduces the affinity of these derivatives for central opioid receptors. They show that this reduction is receptor-type selective, and they extend previous reports demonstrating functional similarities between mammalian and non-mammalian central opioid receptors. PMID- 9200756 TI - Endogenous adenosine facilitates neurotransmission via A2A adenosine receptors in the rat superior colliculus in vivo. AB - The concentration of endogenous adenosine in the cerebrospinal fluid increased 2 3-fold of the original level in the area of rat superior colliculus after the intraperitoneal administration of an adenosine deaminase inhibitor, EHNA (erythro 9-(2-hydroxy-3-nonyl)adenosine, 10 mg/kg). Potentials evoked in the superior colliculus by optic tract stimulation were also facilitated by 120-160% of their initial amplitudes. A selective A1 adenosine receptor antagonist, DPCPX (8 cyclopentyl-1,3-dipropylxanthine), failed to reduce such EHNA-induced facilitation. However, a selective A2A adenosine receptor antagonist, KF17837 (8(3,4-dimethoxystyryl)-1,3-dipropyl-7-methylxanthine) completely eliminated the facilitatory effects of EHNA. Northern blot analysis demonstrated abundant expression of A1 adenosine receptor mRNA in the superior colliculus. RT-PCR analysis was able to detect the concomitant expression of A2A adenosine receptor mRNA, but at levels lower than one-tenth of the striatal expression. In the superior colliculus, A2A adenosine receptors function predominantly on the facilitatory effects of adenosine, irrespective of the ubiquitous expression of A1 adenosine receptors. PMID- 9200758 TI - Operant behavior during sessions of intravenous cocaine infusion is necessary and sufficient for phasic firing of single nucleus accumbens neurons. AB - The activity of individual accumbens neurons in rats was recorded in relation to intravenous cocaine infusions that were either response (i.e., lever press) contingent or response non-contingent. Neural firing was additionally recorded in relation to non-reinforced lever presses. Comparisons of firing under the three conditions showed that operant behavior was necessary and sufficient for preinfusion firing to occur. Surprisingly, the same was true, in many cases, for firing that occurred during the infusion. For other neurons, firing during the infusion was unrelated to operant behavior and possibly related to infusion stimuli. The relationship to operant behavior exhibited by the majority of NAcc neurons is consistent with previous studies that demonstrated a necessary relationship between NAcc neurons and cocaine reinforced operant behavior. PMID- 9200759 TI - Circadian rhythms in mouse suprachiasmatic nucleus explants on multimicroelectrode plates. AB - The suprachiasmatic nucleus (SCN) of the mammalian hypothalamus functions as a circadian pacemaker. This study used multimicroelectrode plates to measure extracellular action potential activity simultaneously from multiple sites within the cultured mouse SCN. Neurons within the isolated mouse SCN expressed a circadian rhythm in spontaneous firing rate for weeks in culture. PMID- 9200760 TI - Differential expression of FGF-2 isoforms in the rat adrenal medulla during postnatal development in vivo. AB - Basic fibroblast growth factor (FGF-2) isoforms of the adrenal medulla are differentially expressed during rat postnatal development. While the 18 and 23 kDa isoforms continuously rise towards the adult expression level, the 21 kDa isoform displays a peak expression at postnatal day 28. The peak expression of the 21 kDa isoform correlates with the peak of the corticosterone concentration during postnatal development. Together with the previously demonstrated increase of the 21 kDa isoform in the adrenal medulla in vivo after glucocorticoid administration these results suggest that the differential regulation of the FGF 2 isoforms could be a physiologically occurring mechanism. PMID- 9200761 TI - Cerebral ischemia and white matter edema in experimental hydrocephalus: a combined in vivo MRI and MRS study. AB - T2 and diffusion weighted MRI, as well as 31P and 1H MRS were performed in kaolin induced hydrocephalic rats. Extracellular white matter edema was detected in the early stages of progressive hydrocephalus. Phosphocreatine (PCr)/inorganic phosphate (Pi) ratios in hydrocephalic animals were decreased compared to controls, and lactate was detected during the acute and chronic stages of hydrocephalus. These MR spectroscopic results are indicative of a compromised energy metabolism and suggest the occurrence of cerebral ischemia in experimental hydrocephalus. PMID- 9200762 TI - The ethics of multiple blood sampling in children for research. PMID- 9200763 TI - The influence of dosage, age, and comedication on steady state plasma lamotrigine concentrations in epileptic children: a prospective study with preliminary assessment of correlations with clinical response. AB - The effects of age, dosage, and type of comedication on plasma lamotrigine (LTG) concentrations and the relationship between plasma drug levels and clinical response were evaluated in a prospective study of 45 patients, aged 3 to 38 years, with epilepsy uncontrolled by conventional anticonvulsant therapy. Six of the 45 patients were on single-drug therapy, and 39 were on two to five concurrently administered antiepileptic drugs when LTG was added. Thirteen patients were assessed at three or more LTG dosage levels. Within individuals, steady state plasma LTG concentrations increased linearly with increasing daily dosage over the examined dose range (25 to 575 mg/day or 0.75 to 21 mg/kg.day). Among patients also receiving enzyme-inducing agents, such as carbamazepine, barbiturates, or phenytoin, plasma LTG concentrations normalized to a 1 mg/kg daily dose were lower in children aged 3 to 6 years (0.30 +/- 0.17 microgram/ml; n = 6) than in the older children (0.43 +/-0.18 microgram/ml; n = 12) and adolescents/adults (0.68 +/- 0.26 microgram/ml; n = 10). In patients treated with valproate, the age dependency of plasma LTG was less evident, possibly because of a smaller sample size and the confounding effect of comedication. Within any given age group, dose-normalized LTG concentrations were about five-fold higher in patients comedicated with valproic acid than in those comedicated with enzyme inducers. Twenty patients showed a favorable response (with a > or = 40% reduction in seizure frequency compared with the pre-LTG period) and continued on long-term treatment. Plasma drug concentrations in these apparent responders were highly variable and did not differ significantly from those observed in nonresponders (6.6 +/- 5.2 versus 4.8 +/- 3.3 microgram/ml). These findings show that plasma LTG concentrations exhibit a wide interindividual variability under the influence of age and type of comedication, but they are predictably related to dosage within individual patients. Although there was no apparent relationship between drug levels and clinical response in this difficult-to-treat population, further studies on the potential value of monitoring plasma LTG concentrations are indicated. PMID- 9200764 TI - Increased plasma concentrations of bromperidol and its reduced metabolite with levomepromazine, but not with thioridazine. AB - Bromperidol is a close structural analog of haloperidol. The authors studied the effects of levomepromazine and thioridazine, which are frequently added to other neuroleptics as sedatives, on plasma concentrations of bromperidol and its reduced metabolite. The subjects were 26 inpatients with schizophrenia receiving bromperidol, 12 to 24 mg/day, for 1 to 19 weeks. In 10 cases, 50 mg levomepromazine per day and in nine cases, 50 mg thioridazine per day were coadministered for 1 week. In seven cases, both drugs were coadministered with > or = 2-week intervals. Plasma concentrations of bromperidol and reduced bromperidol were measured by a high-performance liquid chromatographic method. Levomepromazine (n = 17) significantly (p < 0.001) increased plasma concentrations of bromperidol (7.3 +/- 4.1 versus 10.2 +/- 4.8 ng/ml) and reduced bromperidol (1.8 +/- 1.4 versus 4.5 +/- 3.3 ng/ml). Thioridazine (n = 16) did not significantly change plasma concentrations of bromperidol (9.1 +/- 5.7 versus 8.6 +/- 5.5 ng/ml), while those of reduced bromperidol could not be measured because of interfering peaks. The current study suggests that levomepromazine, but not thioridazine, increases plasma concentrations of bromperidol and reduced bromperidol by inhibiting the metabolism of these compounds. PMID- 9200765 TI - Special considerations for monitoring vancomycin concentrations in pediatric patients. AB - The objectives of this study were to estimate the prevalence of low or excessive vancomycin dosing after initiation of treatment in pediatric patients and to determine the factors that are most predictive of optimized vancomycin dosage in this group. Among 74 patients who underwent vancomycin concentration monitoring, low trough (< 4.0 micrograms/ml) and/or peak (< 15.0 micrograms/ml) concentrations were noted in 28 (38%) patients after the initiation of therapy but in only four of the 28 (14%) patients (p = 0.29) after optimization of the initial dosage. There were not toxic peak concentrations (> 60 micrograms/ml) reported during the study. In patients older than 1 month old, 11 low peaks were associated with troughs less than 7.5 micrograms/ml, whereas no low peaks were associated with troughs more than 7.5 micrograms/ml. The significant predictive variables of optimized vancomycin dosage in the reduced regression model (p < 0.00001; adjusted r2 =0.85; n = 36) were (log) initial dose (p < 0.0001), initial trough (p < 0.0001), and age (p = 0.009). Initial peak concentrations were not associated with the optimized dosage (p = 0.50). The results of this study indicate that approximately 40% of all pediatric patients will be at risk of significant underdosing if standard vancomycin dosing guidelines are followed and that patients older than 1 month old with initial trough concentrations less than 7.5 micrograms/ml are at a greater risk of low peak concentrations than individuals with trough concentrations more than 7.5 micrograms/ml. Monitoring vancomycin concentrations appears to be essential to prevent the underdosing of many pediatric patients; however, only initial trough vancomycin concentrations may be needed to optimize dosages. PMID- 9200766 TI - Amikacin Bayesian forecasting in critically ill patients with sepsis and cirrhosis. AB - This study was designed to determine the population pharmacokinetic parameters of amikacin in two subpopulations of intensive care unit patients with sepsis and cirrhosis and sepsis without cirrhosis. The authors evaluated the usefulness of the obtained parameters to forecast the serum amikacin concentrations in a validation group of patients with sepsis and cirrhosis when used as a priori distribution in a Bayesian forecaster. The population parameters were estimated by a nonparametric expectation maximization algorithm (NPEM), and the accuracy of the predictions were evaluated through a prediction error analysis. Significant differences (p < 0.05) were found in Vd (0.668 versus 0.470 l/kg) and K (0.0701 versus 0.161 h-1) between subpopulations of patients with and without cirrhosis. The model derived for patients with cirrhosis used as a priori distribution, with and without feedback, was superior to the model derived for patients with sepsis in forecasting amikacin serum concentrations. The results show the relevance of using the specific model for the subgroup with cirrhosis as a priori distribution in a Bayesian forecaster to obtain a nonbiased prediction with an acceptable precision. PMID- 9200767 TI - Comparison and validation of limited sampling equations for cyclosporine area under-the-curve monitoring calculations in pediatric renal transplant recipients. AB - Therapeutic monitoring of cyclosporine (CsA) by using area-under-the concentration-time-curve (AUC) values in renal transplant recipients has been previously demonstrated to predict posttransplant clinical outcome. Two previous studies also reported that limited sampling equations could accurately determine the AUC of CsA using one to six blood concentration determinations in adults. The purpose of this study was to validate the accuracy of these equations in a pediatric population. In 18 pediatric patients who received renal allografts, three limited sampling equations, which used one, four, or five concentration time points, accurately estimated CsA AUC when compared with an actual 7- to 9 point curve. An equation that used a single concentration time point at 5 hours was unbiased and provided the best precision in calculating a 12-hour interval AUC. This equation had a mean absolute percentage error of 5.8% (95% confidence interval, 3.3 to 8.3). Equations using four or five concentration time points were found to provide estimates of AUC for a 24-hour interval AUC, with less than 10% error. These findings suggest that limited sampling models using as few as one to four concentration time points provide accurate estimations of CsA AUC in pediatric patients. The use of these limited sampling models provides the clinical advantage of lower blood sampling requirements and reduced costs associated with the monitoring of cyclosporine. PMID- 9200768 TI - Valproic acid-ketoconazole interaction in normal, hypoalbuminemic, and uremic sera: lack of interaction in uremic serum caused by the presence of inhibitor. AB - Ketoconazole is an antifungal agent widely used in the management of patients with fungal infection, especially in patients with acute acquired immuno deficiency syndrome (AIDS). Ketoconazole is 99% bound to serum albumin and may interact with valproic acid, an anticonvulsant with 90% to 95% binding to serum albumin. The interaction may be more significant in hypoalbuminemia, a common finding in patients with AIDS. However, valproic acid-ketoconazole interaction has not been reported. The authors prepared two serum pools from patients receiving valproic acid with normal serum albumin and another pool from patients with hypoalbuminemia. Another serum pool was prepared from uremic patients not receiving valproic acid. The aliquots of serum pool were supplemented with various concentrations of ketoconazole, representing therapeutic and slightly higher therapeutic concentrations. The concentrations of free valproic acid were determined in protein-free ultrafiltrates (prepared by centrifuging specimens at 25 degrees C with the Centrifree Micropartition System at 1500 g for 20 minutes) using fluorescence polarization immunoassay. In the serum pool with normal albumin concentration, the authors observed statistically significant displacement of valproic acid only at higher ketoconazole concentrations (10 and 20 micrograms/ml) whereas, in the serum pool with hypoalbuminemia, they observed statistically significant displacement of valproic acid by ketoconazole with lower and higher concentrations of ketoconazole. The magnitude of displacement was more significant at high valproic acid concentrations (95 and 150 mg/ml, respectively) probably because of the concentration-dependent binding of valproic acid to serum albumin. The authors observed no displacement of valproic acid by ketoconazole in the uremic serum pool. On the other hand, the free valproic acid concentrations were decreased in the presence of ketoconazole in the uremic serum pool. PMID- 9200770 TI - Optimized high-performance liquid chromatographic method for determination of lamotrigine in serum with concomitant determination of phenytoin, carbamazepine, and carbamazepine epoxide. AB - Lamotrigine (LG), phenytoin (PY), carbamazepine (CM), and carbamazepine epoxide (CE) are measured with an optimized procedure that uses thin sorbent extraction disks and a highly selective, sterically protected bonded silica high-performance liquid chromatography (HPLC) column. Routinely, serum (200 microliters at pH 6.8 with cyheptamide as internal standard) is applied to an Empore octyl (C8) solid phase extraction disk to isolate the drugs. a water wash removes interferences, and the retained drugs are eluted with a small volume of solvent. The eluate is directly injected onto a Zorbax Stable Bond cyanopropyl HPLC column with quantification at 214 nm. Evaporation-concentration steps are unnecessary. Overall, for all drugs, between-run precision coefficients of variation (n = 16 each) ranged from 2.1% to 4.9% at concentrations from 0.75 to 20.5 mg/l; extraction recoveries fell within a range of 96% to 110% at concentrations of 2, 10, and 30 mg/l tested for each drug; the lowest limit of detection was 0.15 to 0.35 mg/l. The analytical response was linear for each drug > 80 mg/l (LG) and > 50 mg/l (PY, CM, and CE). Optimization graphs are presented to illustrate the rationale for selection of test parameters for a robust method. In addition, a comparison study between two commercial laboratories demonstrates accuracy problems associated with LG testing. PMID- 9200769 TI - Effect of diltiazem on plasma concentrations of oxypurines and uric acid. AB - To determine the clinical effect of diltiazem on the metabolism of adenosine, and its importance in ischemic heart disease, arterial plasma concentrations of the purine metabolites were determined in 21 healthy volunteers (10 female and 11 male) and 19 patients with effort angina (8 female and 11 male) before, during, and immediately after standard treadmill exercise tests conducted before and after they had taken 60 mg diltiazem (Cardizem; Hoechst Marion Roussel, Laval, QC, Canada) four times a day for 1 week. The results showed that the cardiac patients had significantly lower mean plasma concentrations of uric acid (46.82 +/- 25.51 versus 95.47 +/- 35.41 micrograms/ml, p 0.05), inosine (0.25 +/- 0.19 versus 0.84 +/- 0.17 microgram/ml, p < 0.05), and hypoxanthine (0.28 +/- 0.35 versus 0.50 +/- 0.27 microgram/ml, p < 0.05). Diltiazem decreased the mean resting plasma concentrations of uric acid in patients (uric acid 43.47 +/- 22.26 versus 46.82 +/- 25.51 micrograms/ml, p < 0.05) and healthy volunteers (uric acid 85.68 +/- 26.71 versus 95.47 +/- 35.41 micrograms/ml, p < 0.05). There was no statistically significant change in the plasma concentrations of the purine metabolites during exercise (p < 0.05). Female subjects had significantly lower plasma concentrations of uric acid than males (patients, 34.87 +/- 26.93 versus 55.78 +/- 21.25 micrograms/ml; healthy volunteers, 84.79 +/- 32.07 versus 104.22 +/- 37.05 micrograms/ml; p < 0.05 for both). Results of the study suggest that normal therapeutic doses of diltiazem may modulate the metabolism of adenosine and that some of the purine metabolites may be useful markers for specific types of ischemic heart disease. PMID- 9200771 TI - Determination of felbamate concentration in pediatric samples by high-performance liquid chromatography. AB - A simple and rapid procedure to determine felbamate (2-phenyl-1,3-propanediol dicarbamate) concentrations in human plasma/serum by high-performance liquid chromatography is described. The method employs a high-performance liquid chromatography unit equipped with a C18 reverse-phase cartridge (3-microliters particle diameter, 3.2 x 40 mm), an acetonitrile/water gradient, and detection at 210 nm. The sample is deproteinized with acetonitrile containing internal standard (2-methyl-2-phenyl-1,3-propanediol dicarbamate), and the resulting supernatant, after diluting 1:1 with water, is injected onto the column. The felbamate and internal standard are eluted with a linear gradient of 0% to 22% acetonitrile for 11 minutes at a flow rate of 0.8 ml/minute. Under these conditions, felbamate and the internal standard are eluted at 9.2 +/- 0.03 and 10.8 +/- 0.03 minutes, respectively. The assay is linear from 10 to 400 microgram/ml. It is highly reproducible; at 100 micrograms/ml felbamate, within day and between-day coefficients of variation are less than 0.5% and 4.3%, respectively. Recovery is > or = 95%. No interferences from other common antiepileptic drugs and analgesics are observed. Advantages of this method include simple and fast sample preparation; use of a gradient to eliminate interferences; and use of a cartridge column, which is economical, provides good resolution, allows rapid equilibration and elution, and operates at low back pressures. The method requires samples of only 100 microliters and is ideal for pediatric samples. PMID- 9200772 TI - Plasma level monitoring of olanzapine in patients with schizophrenia: determination by high-performance liquid chromatography with electrochemical detection. AB - A sensitive high-performance liquid chromatography method with electrochemical detection for the determination of olanzapine in human plasma is described. Olanzapine from plasma samples was isolated by a simple one-step liquid--liquid extraction with 15% methylene chloride in pentane with an extraction recovery of approximately 94% of the total olanzapine in plasma. The compound was separated on a cyano column. Under the conditions described, commonly coadministered drugs and other common antipsychotic drugs did not interfere with the analysis of olanzapine. The lower limit of determination of the assay was 0.25 ng of olanzapine per ml when 1 ml of plasma was used for the analysis. The interaassay and intraassay variance was (CV%) less than 10%. The standard curve was linear within the range of 0.25 to 50 ng/ml of olanzapine. This method has been used for the determination of plasma levels of olanzapine in patients with schizophrenia who were treated with daily oral doses of 10, 15, and 20 mg of olanzapine. The results indicate that the plasma level of olanzapine increases linearly with the administered daily oral dose (r = 0.6889, p = 0.01). PMID- 9200773 TI - Carbamazepine: detection of another metabolite in serum, 9 hydroxymethyl-10 carbamoyl acridan. AB - In this article, the authors discuss 9-hydroxymethyl-10-carbamoyl acridan (9-OH CBZ), another metabolite of carbamazepine (CBZ) found in serum. The retention time of unconjugated 9-OH-CBZ should be known when using a chromatographic method, because it appears in concentrations varying from one eighth to one third of the CBZ-10,11-epoxide (CBZ-E) concentration and may therefore cause analytical interactions. Liquid chromatography/electrospray mass spectrometry ((LC/ES-MS) in a serum extract identified and confirmed 9-OH-CBZ. The amount of 9-OH-CBZ present as conjugate in serum was between 42% and 65%. The correlation factor values (r) between serum concentrations of 9-OH-CBZ and 10,11-dihydro-10,11-trans-dihydroxy CBZ (trans-CBZ-diol), CBZ-E, and CBZ in 100 serum samples were 0.77, 0.80, and 0.53, respectively. The origin of 9-OH-CBZ is discussed. PMID- 9200774 TI - Possibilities for therapeutic drug monitoring of azathioprine: 6-thioguanine nucleotide concentrations and thiopurine methyltransferase activity in red blood cells. AB - The objectives of this study were to establish monitoring of azathioprine (AZA) treatment in renal allograft recipients by red blood cell (RBC) 6-thioguanine nucleotide (6-TGN) measurements and to characterize the variability of RBC thiopurine methyltransferase (TPMT) activity and the effects on 6-TGN levels and the incidence of rejection episodes. In 82 renal allograft recipients, the effect of standard AZA dosage (3 mg/kg tapered to 1 mg/kg) was compared with higher dosages (3 mg/kg for several days) under 6-TGN monitoring. The authors measured TPMT in these patients and in a group not receiving AZA. The authors did not find an inverse correlation between RBC TPMT activity and 6-TGN concentrations, and baseline TPMT activity did not predict the incidence of rejection episodes The slight increase in RBC TPMT activity after transplant was associated with the use of furosemide rather than AZA; in the five patients receiving furosemide for less than 10 days, TPMT activity declined. The higher AZA dosage in the 6-TGN monitored group was not sufficient to increase RBC 6-TGN to target levels (100 to 200 pmol/8 x 10(8) RBC); median 6-TGN levels were similar in the two groups, as was the incidence of rejection episodes. Based on these findings, the authors suggest that higher dosages be studied in conjunction with 6-TGN monitoring, to explore the possibilities for therapeutic improvements. PMID- 9200775 TI - A simple and reliable reverse-phase high-performance liquid chromatographic procedure for determination of paclitaxel (taxol) in human serum. AB - A simple, fast, and reliable isocratic (mobile phase: acetonitrile/methanol/water [48/11/41], reverse-phase (C18 column) high-performance liquid chromatography method for the determination of paclitaxel concentration in human serum is presented. The procedure uses a new and convenient one-step sample-purification procedure that requires only 400 microliters of sample and uses N-heptylbenzamide as an internal standard. Paclitaxel is detected by UV absorbance measurement at 227 nm. The method has a broad linear range (0.01 to 10 mg/l, or 0.012 to 11.7 mumol/l; r > 0.999), and the detection limit is 0.01 mg/l (0.012 mumol/l). The deviation from target value is < or = 1.5%, and coefficients of variation are < or = 13.8% within runs and < or = 15.3% between runs. Recovery paclitaxel is > or = 92.6%. No interferences were observed from endogenous compounds or from more than 30 drugs that may be administered with paclitaxel. Docetaxel, which is not concurrently administered, coeluted with paclitaxel. Compared with previously published high-performance liquid chromatography procedures for the determination of paclitaxel, the particular advantage of the method presented here is its simple and rapid single-step sample-purification procedure, which makes a high recovery of paclitaxel from serum samples possible and results in a pure extract, avoiding interferences from endogenous compounds. The method is suitable for pharmacological studies and routine analysis. PMID- 9200776 TI - Therapeutic drug monitoring of risperidone and 9-hydroxyrisperidone in serum with solid-phase extraction and high-performance liquid chromatography. AB - This laboratory developed a simple and efficient solid-phase extraction method that is combined with high-performance liquid chromatography for rapid and precise therapeutic monitoring of risperidone (Risperdal) in blood concentrations. The solid-phase extraction uses a mixed bed column. Sensitivity of the chromatographic method is 0.5 ng/ml (180 pmol/ml) of drug in serum, and separations can be performed in a 15-minute chromatographic run. Advantages of this approach include enhanced speed, sensitivity, and efficiency. A high level of sensitivity may be achieved because of the absence of interference from other drugs, metabolites, or serum components. PMID- 9200777 TI - Measurement of tacrolimus (FK506) and its metabolites: a review of assay development and application in therapeutic drug monitoring and pharmacokinetic studies. AB - Tacrolimus (FK506, Prograf) is a macrolide immunosuppressant used for the prevention of organ rejection after transplantation. Tacrolimus demonstrates considerable interindividual variation in its pharmacokinetic profile. This has caused difficulty in defining the optimum regimen and has highlighted the need for therapeutic drug monitoring. Several assay methods for the measurements of tacrolimus in biological specimens have been developed. These assay methods were used for therapeutic drug monitoring and/or pharmacokinetic studies. Two commercially available immunoassays, based on the same monoclonal antibody to tacrolimus, have been used for therapeutic drug monitoring of tacrolimus in whole blood. For pharmacokinetic studies, the assay methods were used to measure tacrolimus and its metabolites in very low concentrations in selected biological matrixes to determine the metabolic and pharmacokinetic profiles of this drug. PMID- 9200778 TI - A rapid high-performance liquid chromatographic method for the measurement of midazolam plasma concentrations during long-term infusion in ICU patients. AB - A new high-performance liquid chromatographic method was developed for quantification of midazolam in plasma samples from intensive care unit patients on long-term intravenous infusion of this benzodiazepine. Plasma samples (0.5 ml) were mixed with 1 microgram flurazepam (internal standard), alkalinized with 2.5 N NaOH, and extracted with toluene. The organic phase was evaporated to dryness, and the residue was dissolved in the mobile phase (acetonitrile/0.05 M phosphate buffer pH 4.5) and injected into the analytical column (C18 Nova-Pak 3.9 x 150 mm, 4 microns, maintained at room temperature; mobile phase flow rate: 1.2 ml/minute). The eluate was monitored at 207 nm, which reduced the risk of interferences from concurrent medications. Retention times of flurazepam, 1' hydroxymidazolam (an active metabolite) and midazolam were approximately 4.5, 6.1 and 13.5 minutes, respectively. The assay was linear over the range 100 to 3000 ng/ml. The coefficients of variation of the within-day and between-day assay for the 100 to 3000 ng/ml range were < 5% and < 7%, respectively. The developed method is fast, reproducible, and well suited to monitor steady state midazolam plasma concentrations in clinical samples. PMID- 9200779 TI - Effect of mycophenolic acid glucuronide on inosine monophosphate dehydrogenase activity. AB - Mycophenolic acid glucuronide (MPAG) inhibition kinetics were evaluated using purified recombinant human type II inosine monophosphate dehydrogenase (IMPDH). MPAG inhibitory concentrations (IC50) were found to be 532- to 1022-fold higher than those for MPA. As expected, according to tight-binding inhibitor kinetics, mycophenolic acid (MPA) IC50 values increased as IMPDH concentrations increased, whereas IC50 values for xanthosine monophosphate (competitive IMPDH inhibitor used as control), an inhibitor known not to be tight binding, remained independent of enzyme concentration. Although MPAG exhibited only weak inhibition of IMPDH activity, in comparison with MPA, IC50 values increased with increasing enzyme concentration. The presence of trace quantities of MPA (0.2% on a molar basis) in the MPAG preparation, detected by high-performance liquid chromatography analysis, could account for this observation. These data support the proposal that MPAG is a pharmacologically inactive metabolite of MPA. PMID- 9200781 TI - Minimizing digoxin-like immunoreactivity with TDx digoxin in dialysis patients. PMID- 9200780 TI - Decreased hypoprothrombinemic effect of warfarin associated with furosemide. AB - The authors describe a case of furosemide possibly inhibiting the hypoprothrombinemic effect of warfarin. The initiation of furosemide dosing in a patient receiving a stable dose of warfarin was associated with an 28% decrease in the international normalized ration (INR). Using normal volunteers, two previous controlled studies of an interaction between warfarin and chlorthalidone, and between warfarin and spironolactone, assert that volume depletion produced by forced diuresis results in the inhibition of warfarin's hypoprothrombinemia. Consistent with this hypothesis, the authors found that their patient's hematocrit (believed to reflect hydration status) correlated with the INR: r2 = 0.78, p < 0.05. This case provides further evidence suggesting that acute diuresis can decrease the hypoprothrombinemic effect of warfarin. PMID- 9200782 TI - Awareness of potential drug interactions may aid avoidance. PMID- 9200783 TI - Hazardous waste: its impact on human health in Europe. AB - Hazardous waste management is of great concern to the nations of Europe. The European public, like that in North America, expresses great concern that hazardous waste is impacting individual health and degrades the environment. The level of resources and degree of hazardous waste problems varies widely throughout Europe. In particular, the Central and Eastern European countries face enormous challenges in trying to solve their waste problems. Progress in managing the hazardous waste burden is evident in Europe, but cooperation across the nations of Europe will be essential to assure success. PMID- 9200784 TI - Hazardous waste: human health effects. PMID- 9200785 TI - Environmental contamination and health effects: what is the evidence? PMID- 9200786 TI - Ecogenetics: from ecology to health. PMID- 9200787 TI - Lead levels in exposed herring gulls: differences in the field and laboratory. AB - We compared blood lead levels for herring gull (Larus argentatus) chicks exposed at two days of age in the field and the laboratory. One randomly selected chick in each family of three was injected with lead, the second with a sterile saline solution, and the third was not injected. Field birds were then completely free living, and were entirely cared for by their parents. Blood was drawn at 35 or 45 days of age for comparison with laboratory-reared chicks. In both the laboratory and the field, blood lead levels were positively related to dose, and concentrations were lower at 45 than at 35 days of age. However, at each dose, the field birds had lower levels than did the laboratory birds. We postulate that this relates to higher overall activity and accelerated bone development in the field, and perhaps to a move varied diet. Wild young gulls were mobile and practiced flight more often than did laboratory-reared gulls. Growth metabolic, and behavioral factors may enhance deposition of lead in the bone, reducing blood lead. Thus, both in ecological risk assessments and in using birds as bioindicators, caution is required in extrapolating from laboratory studies in field conditions. PMID- 9200788 TI - Biochemical assessment of cyanide-induced toxicity in migratory birds from gold mining hazardous waste ponds. PMID- 9200789 TI - Clinical applications of L-line X-ray fluorescence to estimate bone lead values in lead-poisoned young children and in children, teenagers, and adults from lead exposed and non-lead-exposed suburban communities in the United States. AB - In summary, LXRF estimates of Pb in tibial cortical bone have yielded highly relevant clinical data relating to the efficacy of chelation therapy with CaNa2EDTA in lead poisoned children; diagnostic approach(es) to childhood lead poisoning; and evaluations of exposure in children, teenagers, and adults in lead exposed and non-lead-exposed suburban communities. It is anticipated that KXRF and LXRF estimates of Pb in bone will yield new information concerning the epidemiology of hypertension, osteoporosis, and the contribution of maternal Pb to the developing fetus. It would be premature to delineate the age-developmental time boundaries over which these two systems [KXRF, LXRF] can be viewed as operated optimally. Consequently, it is appropriate to view the two approaches as providing complementary information. All such information might, in fact, be required to obtain a complete exposure profile and a comprehensive framework for assessment of health risks. PMID- 9200790 TI - Performance of cytogenetic biomarkers on children exposed to environmental pollutants. PMID- 9200791 TI - Using measured contaminant concentrations versus modeling for CERCLA-related air pathway risk assessments. PMID- 9200792 TI - Quantifying individual residential exposure to smelter emissions in four Arizona copper smelter communities: exposure estimation procedures and results. PMID- 9200793 TI - Combining qualitative and quantitative approaches to assessing impacts of environments on human health and well-being in local community studies. PMID- 9200794 TI - Science put to service: enhancing environmental health services in communities affected by hazardous substances. PMID- 9200795 TI - Strategies for preventing adolescent mercury exposure in Brazilian gold mining areas. PMID- 9200796 TI - A community-based study of adverse pregnancy outcomes near a large hazardous waste landfill in California. PMID- 9200797 TI - Reported health outcomes among residents living adjacent to a hazardous waste site, Harris County, Texas, 1992. PMID- 9200798 TI - Cancer risk in an arsenic-contaminated area of Chile. PMID- 9200799 TI - The lattice diagram and 2 x 2 tables. PMID- 9200800 TI - Geographic information system (GIS) studies of cancer around NPL sites. PMID- 9200801 TI - The role of demographics in public health assessments at Superfund sites: a case study of Rocky Mountain arsenal. PMID- 9200802 TI - Using patterns of child growth and development to assess communitywide effects of low-level exposure to toxic materials. AB - Child growth is a low tech method for assessing a socially valued health outcome, and its assessment focuses attention on the early portion of the lifespan. It is easily measured and sensitive to a variety of influences including chemical ones. As summary measure of health, it does not signify that a particular chemical is present, but it is a response to a variety of them. When used in conjunction with other epidemiologic methods, child growth assessment constitutes a practical, socially meaningful and biologically reasonable tool to assess the impact on community health and well being of chronic low-level exposures to toxic materials. PMID- 9200803 TI - Local health department activity at hazardous waste sites: a spectrum of responses. PMID- 9200804 TI - Improved data collection and response to surveys with the assistance of neighborhood volunteers in a south Texas city. PMID- 9200805 TI - The process of community involvement--a case study: the Bartlesville, Oklahoma, Lead project. PMID- 9200806 TI - [Burkitt lymphoma as a rare cause of hydronephrosis]. AB - Involvement of the genitourinary tract by lymphoproliferative disorders is rare. Rapid proliferating Burkitt's lymphomas could mimic acute diseases. Due to the extreme chemosensitivity, the lymphoma is cureable and surgery should be done only for histology. Initial surgery should be restricted to acute abdominal complications or complete removal of localised resectable disease. In any case a multiagent chemotherapy is necessary. PMID- 9200807 TI - Anticoagulation with r-hirudin in regular haemodialysis with heparin-induced thrombocytopenia (HIT II). The first long-term application of r-hirudin in a haemodialysis patient. AB - A 69-year-old female patient with renal failure developed heparin-induced thrombocytopenia type II (HIT II) two months after starting haemodialysis therapy with heparin as anticoagulant and a 6-week course of thromboembolism prophylaxis with enoxaparin sodium. The platelet count dropped by 50% as compared with initial values and ex vivo platelet aggregation induced by heparin antibodies (HIPA-test) was detected. Haemodialysis therapy was complicated by a massive thrombosis of dialyzer and ensuing repeated interruptions of treatment. After confirmation of the diagnosis of HIT II haemodialysis therapy was continued with hirudin as anticoagulant. Polysulfone dialyzers and an intravenous bolus of 0.14 mg/kg of recombinant hirudin (r-hirudin) achieved efficient haemodialysis therapy of 4.5 hours, with a minimum therapeutic blood level of hirudin of 0.5 micrograms/mL. More than 50 regular haemodialysis with hirudin anticoagulation were performed without additional problems. The ecarin clotting time (ECT) was used as bedside method to monitor blood levels and for dosage adjustments of hirudin. After the 34th haemodialysis, the frequency (previously 3-4 haemodialyses sessions/week) was reduced to 2 sessions/week. The creatinine clearance increased continuously from initially 2.6 to 10.4 ml/min after the 13th week of hirudin-anticoagulated haemodialysis and the platelet count normalized. In conclusion, we report the first long-term administration of r-hirudin to a patient on regular haemodialysis therapy complicated by heparin-induced thrombocytopenia. The use of hirudin as anticoagulant along with dialyzers impermeable to hirudin offers a novel alternative means of anticoagulation and, even in patients with HIT, enables performing an efficient haemodialysis therapy. Hirudin dosage must be individually adjusted by using bedside drug monitoring of plasma concentrations. PMID- 9200808 TI - Cardiorespiratory consequences to hobble restraint. AB - Mechanical restraints in agitated, violent psychiatric patients are still sometimes in use in the initial phase of emergency treatment, especially when patients are taken to hospital by law enforcement. Sudden death has occurred in persons in hobble restraint. Cardiopulmonary response to prone or upright hobble restraint for three minutes was investigated in six male volunteers in randomised crossover trial. RESULTS: No change was observed in the investigated cardiopulmonary parameters after hobble restraint in the upright position. After hobble restraint in the prone position, mean forced vital capacity decreased by 39.6%, mean forced exspiratory volume by 41%, mean end-tidal carbon dioxide increased by 14.7%, mean heart rate decreased by 21.3%, mean systolic blood pressure decreased by 32.3%, mean diastolic blood pressure decreased by 26.1% and mean cardiac output decreased by 37.4% (P for all reported changes < 0.01). CONCLUSION: Hobble restraint in the prone position leads to a dramatic impairment of hemodynamics and respiration. Upright position and frequent control of vital parameters are necessary to prevent a possibly fatal outcome in persons in hobble restraint. PMID- 9200809 TI - Single-dose pharmacokinetics of teicoplanin during hemodialysis therapy using high-flux polysulfone membranes. AB - Teicoplanin is a new glycopeptide antibiotic with potent activity against Gram positive bacteria. It has been considered to be non-dialyzable due to its high molecular weight (1875-1891 d) and high protein binding (89%). Therefore, a reduced dose was recommended for patients on hemodialysis therapy, with the loading dose being followed by a considerably lower maintenance dose and/or extension of the interval between doses. The present study was performed to evaluate the pharmacokinetics of teicoplanin during hemodialysis therapy using high flux membranes. The pharmacokinetic parameters of teicoplanin were studied in 15 patients with chronic renal failure on hemodialysis. A high flux polysulfone membrane (ultrafiltration coefficient of 40 ml/h/mmHg) was used. Teicoplanin was administered at a dosage of 10 mg.kg-1 body weight in 100 ml isotonic saline solution during the first 10 minutes of hemodialysis therapy. Pharmacokinetic analysis was performed using a three compartment analysis. After a single dose of teicoplanin plasma peak levels were 26.4 +/- 12.0 micrograms/mL (mean +/- SD) after 30 minutes. Teicoplanin concentrations rapidly declined to a nadir of 6.1 +/- 2.5 micrograms/mL at the end of the 3.5-hour session dialysis. Extracorporeal clearance was 39.7 +/- 24.5 mL/min. Removal of 19.3 +/- 7.7% of the drug was estimated if infused during hemodialysis. T 1/2 alpha were 0.37 +/- 0.25 hrs, t 1/2 beta 20.1 +/- 7.1 hrs, and t 1/2 gamma 549.7 +/- 210.5 hrs. We conclude that teicoplanin levels are reduced to a subtherapeutic range during one single high-flux dialysis session if the drug is administered during hemodialysis. Thus, in contrast to previous suggestions relevant amounts of teicoplanin are removed during hemodialysis and thus teicoplanin cannot be viewed as non-dialyzable drug. We recommend obligatory drug monitoring to achieve therapeutic plasma concentrations. PMID- 9200810 TI - Reconstitution of the NF-kappa B system in Saccharomyces cerevisiae for isolation of effectors by phenotype modulation. AB - NF-kappa B is a ubiquitous transcription factor that contributes to the induction of many genes playing a central role in immune and inflammatory responses. The NF kappa B proteins are subject to multiple regulatory influences including post translational modifications such as phosphorylation and proteolytic processing. A very important component of this regulation is the control of their subcellular localization: cytoplasmic retention of NF-kappa B is achieved through interaction with I kappa B molecules. In response to extracellular signals, these molecules undergo degradation, NF-kappa B translocates to the nucleus and activates its target genes. To investigate novel proteins involved in this dynamic response, we have reconstituted the NF-kappa B/I kappa Beta system in the yeast Saccharomyces cerevisiae. We have successively introduced p65, the main transcriptional activator of the NF-kappa B family, which leads to the activation of two reporter genes controlled by kappa B sites, and the I kappa B alpha inhibitory protein, which abolishes this activation. By transforming such a yeast strain with a cDNA library we have performed a genetic screen for cDNAs encoding proteins capable of either dissociating the p65/I kappa B alpha complex or directly transactivating the expression of the reporter genes. The efficiency of our screen was demonstrated by the isolation of a cDNA encoding the p105 precursor of the p50 subunit of NF-kappa B. We also used this system to test stimuli known to activate signalling pathways in yeast, in order to investigate whether the related mammalian cascades might be involved in NF-kappa B activation. We showed that yeast endogenous kinase cascades activated by pheromone, hypo- or hyperosmotic shock cannot modulate NF-kappa B activity in our system, and that the p38 human MAP kinase does not act directly on the p65/I kappa B alpha complex. PMID- 9200811 TI - The linear plasmid pDHL1 from Debaryomyces hansenii encodes a protein highly homologous to the pGKL1-plasmid DNA polymerase. AB - Both the linear plasmids, pDHL1 (8.4 kb) and pDHL2 (9.2 kb), of Debaryomyces hansenii TK require the presence of a third linear plasmid pDHL3 (15.0 kb) in the same host cell for their replication. A 3.5 kb Bam HI-PstI fragment of pDHL1 strongly hybridized by Southern analysis to the 3.5 kb NcoI-AccI fragment of pDHL2, suggesting the importance of this conserved region in the replication of the two smaller pDHL plasmids. The 4.2 kb pDHL1 fragment containing the above hybridized region was cloned and sequenced. The results showed that the cloned pDHL1 fragment encodes a protein of 1000 amino acid residues, having a strong similarity to the DNA polymerase coded for by ORF1 of the killer plasmid pGKL1 from Kluyveromyces lactis. The catalytic and proof-reading exonuclease domains as well as terminal protein motif were well conserved as in DNA polymerases of pGKL1 and other yeast linear plasmids. Analysis of the cloned fragment further showed that pDHL1 encodes a protein partly similar to the alpha subunit of the K. lactis killer toxin, although killer activity was not known in the DHL system. Analysis of the 5' non-coding region of the two above pDHL1-ORFs reveal the presence of the upstream conserved sequence similar to that found upstream of pGKL1-ORFs. The possible hairpin loop structure was also found just in front of the ATG start codon of the pDHL1-ORFs like pGKL1-ORFs. Thus the cytoplasmic pDHL plasmids were suggested to possess a gene expression system comparable to that of K. lactis plasmids. PMID- 9200812 TI - The AFT1 transcriptional factor is differentially required for expression of high affinity iron uptake genes in Saccharomyces cerevisiae. AB - High-affinity iron uptake in Saccharomyces cerevisiae involves the extracytoplasmic reduction of ferric ions by FRE1 and FRE2 reductases. Ferrous ions are then transported across the plasma membrane through the FET3 oxidase FTR1 permease complex. Expression of the high-affinity iron uptake genes is induced upon iron deprivation. We demonstrate that AFT1 is differentially involved in such regulation. Aft1 protein is required for maintaining detectable non-induced level of FET3 expression and for induction of FRE2 in iron starvation conditions. On the contrary, FRE1 mRNA induction is normal in the absence of Aft1, although the existence of AFT1 point mutations causing constitutive expression of FRE1 (Yamaguchi-Iwai et al., EMBO J. 14: 1231-1239, 1995) indicates that Aft1 may also participate in FRE1 expression in a dispensable way. The alterations in the basal levels of expression of the high-affinity iron uptake genes may explain why the AFT1 mutant is unable to grow on respirable carbon sources. Overexpression of AFT1 leads to growth arrest of the G1 stage of the cell cycle. Aft1 is a transcriptional activator that would be part of the different transcriptional complexes interacting with the promoter of the high affinity iron uptake genes. Aft1 displays phosphorylation modifications depending on the growth stage of the cells, and it might link induction of genes for iron uptake to other metabolically dominant requirement for cell growth. PMID- 9200813 TI - Sequence comparison of the Ty1 and Ty2 elements of the yeast genome supports the structural model of the tRNAiMet-Ty1 RNA reverse transcription initiation complex. AB - In the reverse transcription initiation complex of the yeast Ty1 retrotransposon, interaction between the template RNA and primer tRNAiMet is not limited to base pairing of the primer binding site (PBS) with ten nucleotides at the 3' end of tRNAiMet, but three regions named boxes O, 1 and 2.1 interact with the T and D stems and loops of tRNAiMet. Sequence comparison of 33 Ty1 elements and 13 closely related Ty2 elements found in the yeast genome shows that the nucleotide sequence of all elements is highly conserved in the region spanning the PBS and the three boxes. Since the domain of the template RNA encodes a portion of protein TyA, we have calculated its amino acid profile and its nucleotide profile to evaluate the role played by nucleotide sequence conservation in the selection for TyA function and in the maintenance of base pairing interactions for the priming function of Ty1 RNA. Our results show that the nucleotide sequence conservation of Ty1 RNA is constrained not only by selection for Ty1 function but also by maintenance of a given nucleotide sequence able to base pair with the tRNAiMet in the primer-template initiation complex. PMID- 9200814 TI - 'Marker swap' plasmids: convenient tools for budding yeast molecular genetics. AB - One-step gene disruption constructs for disruption of HIS3, LEU2, TRP1 or URA3 with each of the other three markers have been constructed. All of these constructs have been tested and found to effectively convert markers either in gene disruptions or on plasmids. The 'swapped' strains allow the unambiguous genetic analysis of synthetic phenotypes with multiple genes, even if the original gene disruptions were made with the same marker. They also allow introduction of multiple plasmids in a single transformant, even if the original plasmids had the same marker, and allow transformation of plasmids into strains containing gene disruptions made with the same marker that is on the plasmids. These 'marker-swap' plasmids therefore eliminate the need for much subcloning to change markers. Marker-swapped alleles are acceptably stable mitotically and meiotically for most applications. PMID- 9200815 TI - DNA sequencing and analysis of 130 kb from yeast chromosome XV. AB - We have determined the nucleotide sequence of 129,524 bases of yeast (Saccharomyces cerevisiae) chromosome XV. Sequence analysis revealed the presence of 59 non-overlapping open reading frames (ORFs) of length > 300 bp, three tRNA genes, four delta elements and one Ty-element. Among the 21 previously known yeast genes (36% of all ORFs in this fragment) were nucleoporin (NUP1), ras protein (RAS1), RNA polymerase III (RPC1) and elongation factor 2 (EF2). Further, 31 ORFs (53% of the total) were found to be homologous to known protein or DNA sequences, or sequence patterns. For seven ORFs (11% of the total) no homology was found. Among the most interesting protein identification in this DNA fragment are an inositol polyphosphatase, the second gene of this type found in yeast (homologous to the human OCRL gene involved in Lowe's syndrome), a new ADP ribosylation factor of the arf6 subfamily, the first protein containing three C2 domains, and an ORF similar to a Bacillus subtilis cell-cycle related protein. For each ORF detailed sequence analysis was carried out, with a full consideration of its biological function and pointing out key regions of interest for further functional analysis. PMID- 9200817 TI - Molecular cloning of a Candida albicans gene (SSB1) coding for a protein related to the Hsp70 family. AB - We have cloned and sequenced a Candida albicans gene (SSB1) encoding a potential member of the heat-shock protein seventy (hsp70) family. The protein encoded by this gene contains 613 amino acids and shows a high degree (85%) of sequence identity to the ssb subfamily (ssb1 and ssb2) of the Saccharomyces cerevisiae hsp70 family. The transcribed mRNA (2.1 kb) is present in similar amounts both in yeast and germ tube cells of C. albicans. PMID- 9200818 TI - Current awareness on yeast. PMID- 9200816 TI - Sequence analysis of Candida albicans phosphoribosyl-aminoimidazole carboxylase (ADE2) gene. AB - The nucleotide sequence of the Candida albicans ADE2 gene, which encodes phosphoribosylaminoimidazole carboxylase, has been determined. The sequence possesses an uninterrupted open reading frame of 1704 nucleotides corresponding to 568 amino acid residues. The deduced amino acid sequence shares a high degree of homology with ADE2 homologues in other fungal species including Saccharomyces cerevisiae, Pichia methanolica, Schwanniomyces occidentalis and Schizosaccharomyces pombe. Three regions of amino acid sequence were highly conserved among all reported ADE2 genes. The hexanucleotide TGACTC characteristic of genes involved in purine and amino acid biosynthesis is located in front of putative TATA boxes in the promoter region. PMID- 9200819 TI - Quantitative measurement of cell membrane transport: technology and applications. AB - The transport of water and cryoprotective chemicals across cell membranes plays an absolutely fundamental role in the outcome of cryopreservation processing. The diversity of cell types as well as the remarkable range of perturbations that cells are subjected to as part of cryopreservation practices generate many interesting research questions. Simply stated, the extreme conditions typical of cryopreservation protocols extend the limits of membrane transport inquiry well beyond that considered in "normal" cell physiology. This paper provides a brief review of methods which have been used for measuring membrane transport properties, especially those methods developed during the past decade which allow us to measure coupled and uncoupled membrane transport properties of water and cryoprotective agents for individual cells in terms of classical Kedem-Katchalsky membrane transport theory. Representative results obtained from these new technologies will be offered to illustrate their utility and relevance to membrane transport issues arising in cryopreservation practice. Engineers have made significant contributions to this area of research primarily in terms of device development and the application of inverse methods to estimate membrane transport properties. PMID- 9200820 TI - New approaches for studying the permeability of fish embryos: toward successful cryopreservation. AB - This paper describes some new approaches for understanding the permeability of teleost embryos. The dechorionated zebrafish (Brachydanio rerio) was used as a model for basic studies of water and cryoprotectant permeability. These embryos are composed of two compartments, a large yolk (surrounded by the yolk syncytial layer) and differentiating blastoderm cells. Cellular water was distributed unequally in each compartment. Measurements indicated that the total water in the embryo was 74%, while the total water in the yolk was 42%, and total water in the blastoderm was 82%. The internal isosmotic value for the zebrafish embryo is unknown. However, for one-compartment modeling studies of membrane permeability, the mean Lp (+/- SEM) values were 0.022 +/- 0.002 to 0.049 +/- 0.008 microns x min-1 atm-1 at 40 mOsm (assuming this was one possible internal isosmotic value for the entire embryo) and 0.040 +/- 0.004 to 0.1 +/- 0.017 microns x min-1 atm-1 at 300 mOsm (assuming this was another possible internal isosmotic value for the entire embryo). When three- and six-somite embryos were placed in 1.5 and 2.0 M cryoprotectants (dimethyl sulfoxide and propylene glycol), osmometric measurements of volume changes indicated no cryoprotectant permeation. However, similar measurements with methanol revealed a small volume decrease (ca. 8%) and recovery (ca. 5%) for six-somite embryos in a 2.0 M solution. Magnetic resonance (MR) images of the spatial distribution of three cryoprotectants (dimethyl sulfoxide, propylene glycol, and methanol) demonstrated that only methanol permeated the entire embryo within 15 min. The other cryoprotectants exhibited little or no permeation into the yolk over 2.5 h. The results from MR spectroscopy and cryoprotectant microinjections into the yolk suggested that the yolk syncytial layer plays the critical limiting role for cryoprotectant permeation throughout the embryo. PMID- 9200821 TI - Novel microwave technology for cryopreservation of biomaterials by suppression of apparent ice formation. AB - Ice formation inside or outside cells has been proposed to be a factor causing cryoinjury to cells/tissues during cryopreservation. How to control, reduce, or eliminate the ice formation has been an important research topic in fundamental cryobiology. The objective of this study was to test a hypothesis that the coupled interaction of microwave radiation and cryoprotectant concentration could significantly influence ice formation and enhance potential vitrification in cryopreservation media at a relative slow cooling rate. Test samples consisted of a series of solutions with ethylene glycol (a cryoprotectant) concentration ranging from 3 to 5.5 M. A specific microwave resonant cavity was built and utilized to provide an intense oscillating electric field. Solutions were simultaneously exposed to this electric field and cooled to -196 degrees C by rapid immersion in liquid nitrogen. Control samples were similarly submerged in liquid nitrogen but without the microwave field. The amount of ice formation was determined by analysis of digital images of the samples. The morphology of the solidified samples was observed by cryomicroscopy. It was found that ice formation was greatly influenced by microwave irradiation. For example, ice formation could be reduced by roughly 56% in 3.5 M ethylene glycol solutions. An average reduction of 66% was observed in 4.5 M solutions. Statistical analysis indicated that the main effects of microwave and ethylene glycol concentration as well as the interaction between these two factors significantly (P < 0.01) influenced ice formation amount, confirming the hypothesis. This preliminary study suggests that a combined use of microwave irradiation and cryoprotectant might be a potential approach to control ice formation in cells/tissues during the cooling process and to enhance vitrification of these biomaterials for long term cryopreservation. PMID- 9200822 TI - Minimally invasive cryosurgery--technological advances. AB - The technological advances which have caused renewed interest in cryosurgery are the development of intraoperative ultrasound to monitor the therapeutic process and the development of new cryosurgical equipment designed to use supercooled liquid nitrogen. The thin, highly efficient probes, available in several sizes, can be placed in diseased sites via endoscopy or percutaneously in minimally invasive procedures. The manner of use is to place the probe in the desired location in the diseased tissue with ultrasound guidance. If required by the size or location of the tumor, as many as five probes can be inserted and cooled to 195 degrees C simultaneously. The process of freezing is monitored by ultrasound which displays a hypoechoic (dark) image when the tissue if frozen. Rapid freezing, slow thawing, and repetition of the freeze/thaw cycle are standard features of technique. Clinical applications which have become common in the past 4 years include the treatment of prostatic cancer and liver tumors. The cases selected for cryosurgery are generally those for which no conventional treatment is possible. However, especially in prostatic cancer, the operative morbidity is so low and the results of therapy are sufficiently good in the short term to merit consideration of use in earlier stages of the disease. Diverse tumors in other sites, such as the brain, bronchus, bone, pancreas, kidney, and uterus, have also been treated in small numbers by cryosurgery. Judging from this experience, further expansion in the use of cryosurgical techniques seems certain. PMID- 9200823 TI - Quail egg yolk: a novel cryoprotectant for the freeze preservation of Poitou jackass sperm. AB - For many years, attempts have been made to establish a sperm bank for the Poitou jackass population which is threatened with extinction. Unfortunately, no cryopreservation technique has ever been described for spermatozoa of this species. In an attempt to find a suitable technique, we studied the relative effectiveness of chicken egg yolk and quail egg yolk in preserving the motility and characteristics of movement of Poitou jackass spermatozoa during the freezing thawing process. Semen was diluted to 60 x 10(6) sperm/ml in a preservation medium containing 4% (v/v) glycerol with 0, 2, 5, 10, 15, or 20% (v/v) of chicken or quail egg yolk. The chemical composition of these two eggs was compared. Effects were assessed using an automated analyzer which measured curvilinear velocity (VCL), straight line velocity (VSL), and the velocity of the average path. Linearity was defined as VSL/VCL x 100. The amplitude of the lateral head displacement was also measured. It was found that after the freeze-thaw process, quail egg yolk improved the percentages of motile and progressively undulating spermatozoa and the movement characteristics compared with chicken egg yolk. The optimal concentration of quail egg yolk was 10%. The general composition of the two types of egg yolk were similar, but quail egg yolk contained significantly more phosphatidylcholine, less phosphatidylethanolamine, and a smaller ratio of polyunsaturated to saturated fatty acids than chicken egg yolk. The improvement of motility for frozen-thawed Poitou jackass spermatozoa using frozen-thawed quail egg yolk compared to chicken egg yolk may be due to the differences in composition of the two yolks. PMID- 9200824 TI - Gross damage accumulation on frozen rabbit liver due to mechanical stress at cryogenic temperatures. AB - The second phase of a pilot study dealing with the mechanical response of frozen biological tissues to external compressive load is presented. This stage deals with histological observations of the damage accompanying mechanically induced permanent deformation in frozen rabbit liver. no significant gross histological damage was observed in the liver samples due to either processing the tissue in the frozen state, due to slow cooling of the liver tissues down to -20 degrees C, or due to rapid cooling of the samples down to -196 degrees C. No histological changes were observed in tissue samples that were loaded within the elastic regime, that is, below the yield strength of the material. Therefore, it is concluded that histological changes due to mechanical stresses are associated with plastic (permanent) deformations. Histological observations indicate that linear cracks which appear to have no preferred orientation develop due to mechanical stress beyond the yield strength of the frozen tissue. These cracks accumulate until final failure of the frozen tissue, when the tissue sample collapses to rubble. Based on histological observations and concepts from solid mechanics, an interaction between crack formation and irregularities in the frozen medium is suggested. Significant sources for such irregularities, in an homogeneous tissue such as the liver, are blood vessels and bile ducts. These irregularities may either initiate crack formation or, on the other hand, may also arrest propagating cracks. PMID- 9200825 TI - The efficacy of antioxidants administered during low temperature storage of warm ischemic kidney tissue slices. AB - Accumulation of products of lipid peroxidation (malondialdehyde, conjugated dienes, lipid peroxides, and Schiff bases) was evaluated in rabbit kidney cortex slices made ischemic for 60 min followed by 18 h storage at 5 degrees C in UW Na gluconate solution and 210 min normothermic reoxygenated incubation. In addition, the effect of adding Trolox (1 mM), deferoxamine (1 mM), and ascorbate (1 mM) as supplemental antioxidants to the UW gluconate solution was evaluated. Lipid peroxidation was slightly increased after hypothermic storage compared to slices subjected to ischemia alone but was not significantly different than ischemic slices during subsequent incubation at normothermia. The addition of either deferoxamine or Trolox to the storage solution substantially reduced lipid peroxidation both during hypothermic storage and subsequent to normothermic incubation. Ascorbate had a mild prooxidant effect as a sole additive to the UW gluconate solution but was clearly prooxidant when combined with either deferoxamine or Trolox. These results suggest that supplemental antioxidants added to the UW gluconate solution under conditions analogous to machine perfusion preservation have a potential role in reducing oxidative stress in kidney tissues harvested after warm ischemia and that hypothermia may be a valuable adjunct to resuscitative therapeutic regimens developed for salvage of ischemic kidneys for transplantation. PMID- 9200826 TI - Postglacial genetic differentiation of reproductive ecotypes of kokanee Oncorhynchus nerka in Okanagan Lake, British Columbia. AB - Okanagan Lake, south-central interior of BC, contains two reproductive ecotypes of kokanee Oncorhynchus nerka; individuals spawn in tributary streams ('stream spawners') as well as on shoreline gravel areas ('beach-spawners'). We tested the hypothesis that these sympatric ecotypes comprise a single panmictic population by assaying variation in morphological traits and at allozyme, mitochondrial and minisatellite DNA loci in fish collected from three stream-spawning and two beach spawning sites. No morphological traits consistently distinguished the reproductive ecotypes with the exception of the number of anal fin rays which was greater in stream-spawning kokanee. Four of 18 allozyme loci screened were polymorphic, but no significant allele frequency differences were detected among populations within ecotypes or between ecotypes. Similarly, allele frequencies at two minisatellite DNA loci were not significantly different among populations or between ecotypes. By contrast, significant differences in the frequencies of mitochondrial DNA restriction fragment length polymorphism (mtDNA RFLP) haplo types were detected between stream- and beach-spawners, but not among populations within ecotypes. Further, two RFLPs that distinguished stream- and beach-spawning adults were found in juvenile kokanee sampled from the limnetic zone of Okanagan Lake. The two mtDNA RFLPs and a d-loop sequence variant appear to be unique to Okanagan Lake Kokanee because we did not observe these haplotypes in sockeye salmon and kokanee sampled outside of Okanagan Lake. Our data suggest that: (i) there is restricted female-mediated gene flow between stream- and beach-spawning kokanee in Okanagan Lake, (ii) the forms have diverged within the lake basin since the retreat of the Wisconsinian glaciers (< approximately equal to 11 000 years ago), and (iii) distinct reproductive niches may promote divergence in north temperate freshwater fish faunas. PMID- 9200828 TI - Mitochondrial DNA sequence variation and phylogeography among Salmo trutta L. (Greek brown trout) populations. AB - To investigate the phylogenetic relationships and geographical structure among brown trout S. trutta L. Populations from the South Adriatic-Ionian and Aegean sea basins, mitochondrial DNA sequence comparisons were used. A 310-base-pair (bp) segment of the control region (D-loop), and an additional 280-bp segment of the cytochrome beta gene were sequenced from representatives of 13 brown trout populations. Phylogenetic analyses, conducted after combining the data presented with published data from other Eurasian brown trout, revealed four major phylogenetic groups, three of which were found widely distributed within the southern Balkan region. The phylogeographical patterns revealed by mtDNA represent one of the few cases where phylogenetic discontinuity in a gene tree exists without obvious geographical localization within a species' range and has most likely resulted from the differentiation of the major mtDNA clades during Messinian or early Pleistocene times. Finally, the genetic relationships among the populations suggested by mtDNA were generally not in accordance with either allozyme or morphological data. PMID- 9200830 TI - Detection and isolation of nuclear haplotypes by PCR-SSCP. AB - SSCP (single-strand conformational polymorphism) is used widely in the field of human biomedicine, but its potential as a population genetics tool for the recovery of nuclear gene genealogies remains to be realized. We describe and illustrate a use for SSCP in the physical isolation of nuclear haplotypes that circumvents several difficulties associated with more conventional cloning procedures. The DNA sequence can be determined directly from the isolated haplotypes and used for phylogenetic inference. SSCP provides a convenient first step toward generating nuclear genealogies for population studies. PMID- 9200831 TI - Characterization of microsatellite loci in the greater white-toothed shrew Crocidura russula. PMID- 9200832 TI - Isolation and characterization of microsatellite loci from Apodemus flavicollis (Rodentia, Muridae) and Clethrionomys glareolus (Rodentia, Cricetidae). PMID- 9200833 TI - Extensive maternal representation of DNA-binding proteins that interact with regulatory target sites of the Strongylocentrotus purpuratus CyIIIa gene. AB - Nine different embryonic transcription factors interact at specific target sites in the cis-regulatory domain of the Strongylocentrotus purpuratus CyIIIa gene. We tested eight of these site sequences and show here that for every one a DNA binding protein was present in unfertilized egg cytoplasm. The concentrations of active DNA-binding proteins per egg were estimated. We also present a new and convenient method for preparation of a material cytoplasmic fraction that retains these factors in active form. PMID- 9200834 TI - Delayed in vitro fertilization of zebrafish eggs in Hank's saline containing bovine serum albumin. AB - In zebrafish it is possible to create viable diploid fish whose genomic DNA is derived only from the female parent (parthenogenesis) or, as was more recently shown, only from the male (androgenesis). Androgenesis requires holding zebrafish eggs in an inactivated state in vitro for an hour or more. Previously this was achieved by placing the zebrafish eggs in ovary fluid obtained from rainbow trout (Onchorhynchus mykiss) or coho salmon (Onchorhynchus kisutch). Here we report that adding bovine serum albumin (BSA) to Hank's buffered saline prevents zebrafish egg activation in vitro. Of the zebrafish eggs placed in Hank's saline plus 0.5% BSA, 85% +/- 8.7% were fertilizable after incubation for one hour at room temperature (23 degrees C). Longer incubations are possible but with lower efficiency of fertilization. This technique not only could facilitate androgenesis, but also might be useful when making transgenics by microinjection, when performing antibody or RNA injections before fertilization, or for studying the mechanisms of egg activation in zebrafish. PMID- 9200836 TI - Efficient gene transfer into zebrafish skeletal muscle by intramuscular injection of plasmid DNA. AB - The ability of zebrafish skeletal muscles to internalize and express plasmid DNA was demonstrated using pCMVCAT1, a chloramphenicol acetyltransferase (CAT) construct driven by the human cytomegalovirus immediate early (CMV-IE) promoter. We found that CAT activity was correlated to the amount of plasmid DNA injected, with maximal expression at 5 micrograms of pCMVCAT1. CAT activity was also shown to increase steadily over the first seven days after injection, with high levels of CAT expression persisting up to one year. Intramuscular injection of CAT constructs driven by other viral promoters also resulted in high levels of CAT activity. Histochemical localization using a CMV beta-galactosidase construct confirmed that only myofibers at the site of injection expressed beta galactosidase enzyme. The persistence and strong expression of injected plasmid constructs suggest that zebrafish may be a simple and readily accessible system for direct muscle injection studies. PMID- 9200835 TI - Zebrafish (Danio rerio) p53 tumor suppressor gene: cDNA sequence and expression during embryogenesis. AB - Three methods were used in succession to screen a whole adult zebrafish cDNA library for expressed p53-like genes. The sequences of the resultant clones describe an open reading frame 1122 nucleotides in length, with another 43 and 940 bases of 5' and 3' untranslated sequence, respectively. The deduced amino acid sequence of the zebrafish p53 protein is 63% identical to that of trout and 48% identical to that of human p53. Two of the three zebrafish clones overlap to span the entire reported cDNA sequence and are identical in their deduced amino acid sequence over their coincident length. The third clone contains a conservative amino acid change, as well as an inserted amino acid subsequently found to be at the junction of exons 2 and 3, suggestive of alternative splicing in the p53 mRNA for this species. Northern analysis demonstrated a zebrafish p53 related transcript to be present and most abundant in zygotes and early-cleavage embryos less than 1 hour after fertilization, thereafter declining to barely detectable levels at 48 hours. A similar temporal expression was detected for the zebrafish L-myc, known to be present in maternally derived RNA, whereas zebrafish N-myc and the zebrafish homologue of the murine T gene were not detectable prior to the onset of zygotic transcription. PMID- 9200837 TI - Random amplified polymorphic DNA (RAPD) markers for determination of genetic variation in wild populations of the black tiger prawn (Penaeus monodon) in Thailand. AB - Random amplified polymorphic DNA (RAPD) analysis was used to amplify the genome of black tiger prawns (Penaeus monodon) to detect DNA markers and assess the utility of the RAPD method for investigating genetic variation in wild P. monodon in Thailand. A total of 200 ten-base primers were screened, and 84 primers yielded amplification products. Six positive primers that gave highly reproducible RAPD patterns were selected for the analysis of three geographically different samples of Thai P. monodon. A total of 70 reproducible RAPD fragments ranging in size from 200 to 2000 bp were scored, and 40 fragments (57%) were polymorphic. The RAPD analysis of broodstocks from three different locales, Satun Trang, Trat, and Angsila, revealed different levels of genetic variability among the samples. The percentages of polymorphic bands were 48% and 45% in Satun-Trang and Trat, respectively, suggesting a high genetic variability of the two samples to be used in selective breeding programs. Only 25% polymorphic bands were found in the Angsila sample, indicating the lowest polymorphic level among the three samples examined. Primer 428 detected a RAPD marker that was found only in P. monodon originating from Satun-Trang, suggesting the potential use of this marker as a population-specific marker in this species. PMID- 9200838 TI - Characterization of gene expression of a p53 homologue in the soft-shell clam (Mya arenaria). AB - Expression of a clam p53 homologue was examined in tissues of the soft-shell clam, Mya arenaria, from Beal's Island, Maine. Southern analysis reveals that p53, in this population, is a single copy gene. A 1.7 to 1.9-kb p53 mRNA was detected at very low levels in normal adult gonadal tissue. This transcript is similar in size to that of vertebrate p53 genes. RNAs were harvested from several tissues, including individual clam gonads during gametogenesis. These were hybridized in ribonuclease (RNase) protection assays to a p53 antisense probe designed from the clam p53 cDNA sequence. RNase protection profiles indicate that p53 mRNA is expressed in adductor muscle, gill, and gonads of both sexes. Although p53 mRNA is expressed throughout gametogenesis in mature male and female gonads, ovaries have significantly higher levels of expression. The significance of our findings to the study of normal clam gametogenesis and to etiology of gonadal tumors is discussed. PMID- 9200839 TI - Intron-length polymorphism at the actin gene locus mac-1: a genetic marker for population studies in the marine mussels Mytilus galloprovincialis Lmk. and M. edulis L. AB - A novel intron-length polymorphism at the actin gene locus mac-1 is here reported and used as a genetic marker for population studies in mussels of the genus Mytilus. Two closely related genes subsequently identified as alleles, mac-1a1 and mac-1b1, from a genomic library of M. galloprovincialis were partially cloned and sequenced. They mainly differed from each other by a 65-bp insertion within their first intron. Polymerase chain reaction (PCR) primers were designed outside the insertion. The PCR analysis of 166 individual mussels from M. galloprovincialis and M. edulis populations revealed three size-classes of alleles or allelomorphs, two of which were of the expected sizes for mac1a1 and mac-1b1. One allelomorph was absent from M. edulis samples, although it was present at substantial frequencies in M. galloprovincialis populations. The frequencies of the two other allelomorphs significantly differed between M. galloprovincialis and M. edulis populations. The comparison of six mac-1 intron sequences over 277 bp showed at once that allelomorphs encompassed alleles differing from one another by substantial numbers of mutations, and that identical alleles were present in both M. galloprovincialis and M. edulis individuals, a probable result of the recent introgression between the two species. PMID- 9200840 TI - Amplification of the complete mitochondrial genome of two protostome worms: a useful technique for comparative studies of metazoan mitochondrial DNA. AB - We report the first polymerase chain reaction (PCR) amplification of the complete mitochondrial genomes of a nemertean and a sipunculan worm in one piece using a recently published two-polymerase protocol for long and accurate DNA amplification. Successful amplification was achieved from nanogram quantities of both purified mitochondrial DNA (nemertean) and crude total DNA (sipunculan). This technique allows the rapid generation of sufficient quantities of entire mitochondrial DNAs for cloning and restriction fragment length polymorphism (RFLP) analyses, and thus will facilitate comparative studies of metazoan mitochondrial genomes. PMID- 9200842 TI - The urinary excretion of solvents and gases for the biological monitoring of occupational exposure: a review. AB - 'In the field' application of the measurement of urinary excretion of unmodified solvent for the biological monitoring of exposed workers has been investigated in many recent papers. The results obtained for several solvents are reviewed. The values of correlation coefficients (r) and regression lines obtained for benzene, toluene, xylene, styrene, n-hexane, cyclohexane, 2- and 3-methylpentane, methyl chloride, tetrachloroethylene, carbon tetrachloride, methyl chloroform, p dichlorobenzene, nitrous oxide, halothane, isoflurane, enflurane, acetone, methyl ethyl ketone and methyl isobutyl ketone are presented. The correlations observed were generally good: r values range from 0.50-0.97, and the majority are between 0.84 and 0.90. The regression lines reported for the same solvent in different studies present some variability: this is possibly due to an inadequate control of factors influencing the relationship between external dose and absorption, such as differences in body burden, work load, individual characteristics, etc. These factors are discussed. As a whole, results reported in the literature show that measuring of urinary excretion of unmodified solvents provides a highly sensitive and specific exposure index, and can also be applied for the biological monitoring of occupational exposure to low levels of solvents or to solvent mixtures. Nevertheless, for an adequate assessment of biological limit values, further studies evaluating the reproducibility of regression lines are needed, given that the aspects influencing the correlation between external dose and urinary excretion are fully controlled. Another crucial aspect is the correlation with early effects: even though this has yet to be evaluated for several solvents, for others such as styrene and perchloroethylene a good correlation was obtained, further supporting the usefulness of the measurement of urinary excretion of solvent for the biological monitoring of occupational exposure. PMID- 9200841 TI - Temporal expression pattern of insulin-like growth factor mRNA during embryonic development in a teleost, rainbow trout (Onchorynchus mykiss). AB - The appearance of insulin-like growth factor I (IGFI) and II (IGF-II) mRNA was studied during rainbow trout embryonic development. Reverse transcription polymerase chain reaction (RT-PCR), followed by Southern blotting and high stringency hybridization with rainbow trout IGF-I and IGF-II cDNA probes, was used to detect all four forms of IGF-I mRNA and one form of IGF-II mRNA from whole-embryo total RNA isolated from a staged series. IGF-I and IGF-II mRNA were detected in unfertilized eggs. IGF-I and IGF-II mRNA were also detected during cleavage, gastrulation, and organogenesis, at hatching, during yolk absorption, and at feeding. IGF-IEa-1 and Ea-3 mRNA were detected in unfertilized eggs, while IGF-I Ea-4 was first detected at stage 9 when the zygotic genome is believed to become activated in rainbow trout. IGF-IEa-2 was not detected at any stage of embryonic development. IGF mRNA of immunoreactive peptides have been detected durin embryonic development in all vertebrates studied to date. Our results confirm the presence of IGF-I mRNA in teleost embryos. In addition, IGF-II mRNA is also present in teleost embryos. Our results suggest that the role IGF-I and IGF-II play during embryonic development may be conserved in vertebrates. PMID- 9200843 TI - Excretion of N-acetyl-S-(1-phenyl-2-hydroxyethyl)-cysteine and N-acetyl-S-(2 phenyl-2-hydroxyethyl)-cysteine in workers exposed to styrene. AB - Styrene (S) has been shown to be responsible for neurotoxic effects, including behavioural changes and neuroendocrine disturbances. The initial step of S metabolism is conversion to styrene 7,8-epoxide (SO), which is present in two enantiomeric forms [(R)(+)-SO and (S)(-)-SO]; this electrophilic intermediate is considered to be directly responsible for most toxic effects of S. The major urinary metabolites derived from the biotransformation of SO in man are mandelic acid (MA) and phenylglyoxylic acid (PGA). In rats an alternative pathway has been demonstrated, which involves the conjugation of SO to glutathione (GSH), leading to the excretion of two specific mercapturic acids, N-acetyl-S-(-(1-phenyl-2 hydroxyethyl)-cysteine [M1] and N-acetyl-S-(2-phenyl-2-hydroxy-ethyl)-cysteine [M2]; a close relationship has been found between exposure to S and urinary excretion of M1 and M2 in rats. As a consequence of the chiral nature of SO, both M1 and M2 consist of two diastereoisomers (M1-'R', M1-'S', M2-'R' and M2-'S'). Early reports have shown that the conversion of S to mercapturic acids is much lower in man (below 1% of the absorbed dose) than in rats (about 10%). We propose an analytical method for the determination of urinary M1 and M2 in man, which involves a urine clean-up by a chromatographic technique with a short reversed phase pre-column; purified samples are then deacetylated with porcine acylase and deproteinized by centrifugal ultrafiltration. A derivatization is then performed with o-phthaldialdehyde and 2-mercaptoethanol and the fluorescent derivatives are separated on a reversed-phase analytical column. The mobile phase consists of acetate buffer and methanol mixed at variable proportions, the fluorescence detector is set at 330 nm (exc.) and 440 nm (em.). M1-'S' and M1-'R' are separated (retention times = 52.8 and 73.7 min, respectively) while the diastereoisomers of M2 coelute as a single peak at 70.5 min. The detection limit is about 7 micrograms/l, the coefficients of variation are below 7% and the error percentages are less than 6%. The method was applied to 25 urine samples from workers exposed to S: significant correlations were found between mercapturic acids and MA and PGA, the best correlation being between M2 and PGA (r = 0.79). Urine samples form unexposed subjects showed no detectable amounts of the analytes. A high stereoselectivity is shown by the enzymes involved in the metabolism of S to mercapturic acids: M1-'S', which derives from (S)-SO, is excreted in much higher amounts than M1-'R', which derives from (R)-SO. PMID- 9200844 TI - A biological monitoring study of 1-methoxy-2-propanol: analytical method development and a human volunteer study. AB - 1-Methoxy-2-propanol (M2P) is finding increasing industrial use as a less toxic alternative to the short-chained ethylene glycol ethers. Like most glycol ethers, M2P is readily absorbed through the skin and biological monitoring is therefore appropriate in assessing occupational exposure. An analytical method, suitable for routine monitoring, was developed for the determination of free M2P in urine. The method involves solvent extraction, gas chromatography-mass spectrometry and is sensitive (detection limit 1 mumol/l), specific and reproducible (intra- and inter-assay coefficients of variation 5% and 9%, respectively). A human volunteer study, involving six volunteers, was also conducted. Volunteers were exposed to 100 ppm M2P for 8 h (the occupational exposure standard in the UK) including a 30 min break. Post-exposure levels of free M2P in urine were found to reach up to 110 mumol/l). Levels of M2P were also monitored in blood (maximum 103 mumol/l) and exhaled air samples (up to 252 nmol/l). The volunteer study showed that M2P is rapidly excreted in urine with a half-life of less than 2.6 h. PMID- 9200845 TI - Biomonitoring of technical grade 1-alkoxy-2-propanol acetates by analysing urinary 2-alkoxypropionic acids. AB - Today commercially available 1-alkoxy-2-propanol acetates contain a harmful isomer 2-alkoxy-1-propanol acetates, which makes up less than 20% of the commercial product. This harmful isomer provides a means with which to study the toxicity and exposure to alkoxypropanols. Assessing exposure to technical grade 1 alkoxy-2-propanol acetates through the biological monitoring of urinary 2 alkoxypropionic acids has been found to be the most accurate method. The method developed in this study provides a procedure for the simultaneous urinalysis of methoxyacetic acid (MAA), ethoxyacetic acid (EAA), butoxyacetic acid (BAA), oxalic acid (OA), 2-methoxypropionic acid (2-MPA) and 2-ethoxypropionic acid (2 EPA). This possibility is very valuable in workplaces where workers are exposed simultaneously to different glycol ethers. This study was conducted among 54 silkscreen printers, who gave a urine sample to be analysed using a capillary gas chromatograph for 2-MPA and 2-EPA. The mean urinary concentrations of 2-MPA and 2 EPA were 1.27 (S.D. = 1.60) nmol/mol creatinine (median = 0.53, n = 26) and 1.23 (S.D. = 2.31) mmol/mol creatinine (median = 0.26, n = 39), respectively. The urinary excretion of 2-MPA and 2-EPA immediately after shift was linearly dependent on the preceding technical grade 1-methoxy-2-proponol acetate (7 = 0.16x + 0.26, n = 26 R2 = 0.78) and technical grade 1-ethoxy-2-propanol acetate (y = 2.05x - 0.09, n = 39, R2 = 0.68) respective exposure, as measured in the workers' breathing zone. According to the results of this study it is possible to monitor exposure to the technical grade 1-methoxy-2-propanol acetate and technical grade 1-ethoxy-2-propanol acetate through urinalysis of 2-MPA and 2 EPA. PMID- 9200846 TI - Inter-individual variability of benzene metabolism to trans,trans-muconic acid and its implications in the biological monitoring of occupational exposure. AB - Unmodified benzene (UBz) and trans,trans-muconic acid (t,t-MA) were measured in urine samples collected at the end of the first half-shift in 80 bus drivers from a large city in Northern Italy. Mean UBz was 1155 ng/l (S.D. = 494), range 85 1980 ng/l; these values roughly correspond to 10-1000 micrograms/m3 of benzene in air. Mean t,t-MA was 297 micrograms/g creatinine; the range was large (20-1295 micrograms/g creatinine), and the distribution of values was bimodal. At further analysis of t,t-MA data, two subgroups of 59 and 18 subjects were identified (3 outliers were excluded): mean values of the index were 108 (S.D. = 65) and 916 (S.D. = 264) micrograms/g creatinine respectively, and the values within each subgroup were normally distributed. The mean ratio between t,t-MA and UBz in the subgroups were 0.15 and 0.85, respectively; the difference was significant. The first subgroup was defined as 'poor t,t-MA metabolizers', the other as 'efficient t,t-MA metabolizers'. No inter-subgroup differences were observed regarding the main characteristics (age, dietary and smoking habits, etc.). As the parent compound of t,t-MA, trans,trans-muconaldehyde is myelotoxic, and its production has been implicated in benzene-induced leukemia. 'efficient' t,t-MA metabolizers may be at higher risk of developing benzene toxicity. If confirmed in further studies, the inter-individual variability rate of metabolizing benzene to t,t-MA may introduce some limitations in the application of this metabolite as an exposure index of low benzene exposure. Nevertheless, the t,t-MA/UBz ratio may be an important index of susceptibility to benzene toxicity. PMID- 9200848 TI - Inhalation toxicokinetics of 1,2,4-trimethylbenzene in volunteers: comparison between exposure to white spirit and 1,2,4-trimethylbenzene alone. AB - The objective of this study was to compare the toxicokinetics of inhaled 1,2,4 trimethylbenzene (1,2,4-TMB) in man after exposure to white spirit with that observed after exposure to 1,2,4-TMB alone. TMBs occur mainly in petroleum products and the TMBs or their metabolites have been suggested as suitable biomarkers of exposure to white spirit and other distillation products. The toxicokinetics were studied in 9 male, healthy volunteers exposed to solvent vapours in an exposure chamber for 2 h during a work load of 50 W. The subjects were exposed to 11 mg/m3 of 1,2,4-TMB on two occasions; during exposure to 1,2,4 TMB vapour alone and during exposure to 300 mg/m3 of white spirit. The 1,2,4-TMB isomer was analyzed in blood and exhaled air by gas chromatography. In addition, a major urinary metabolite of 1,2,4-TMB, 3,4-dimethylhippuric acid (3,4-DMHA), was analyzed by high performance liquid chromatography. Further the occurrence of acute effects was studied by means of a questionnaire. Irritation and central nervous system symptoms were recorded by ratings on a 100-mm visual analogue scale. Blood levels of 1,2,4-TMB and excretion rates of 3,4-DMHA in urine were markedly elevated both during and after exposure to white spirit as compared to exposure to TMB alone. Thus, it appears that components in white spirit inhibit the metabolic elimination of 1,2,4-TMB. This should be considered in biological exposure monitoring as well as in risk assessment. No irritation or central nervous system effects were reported at these conditions. PMID- 9200847 TI - Biological monitoring of exposure to benzene in the production of benzene and in a cokery. AB - The purpose of this study was to compare different biological methods in current use to assess benzene exposure. The methods involved in the study were: benzene in blood, urine and exhaled air, and the urinary metabolites t,t-muconic acid (MA) and S-phenylmercapturic acid (S-PMA). Blood, urine and exhaled air samples were collected from workers in a benzene plant (pure benzene exposure) and cokery (mixed exposure, e.g. polycyclic aromatic hydrocarbons--PAHs) in an Estonian shale oil petrochemical plant. The benzene in these samples was analysed with a head-space gas chromatograph, and the metabolites MA and S-PMA with a liquid chromatograph using methods developed from published procedures. Some of the values measured in the Estonian shale oil area were high in comparison with those published during the last few years, whereas the values measured in the control group did not show any exposure to benzene except in the smokers group. The highest median exposure was in the benzene factory, 0.9 cm3/m3 TWA (2.9 mg/m3) and the highest individual value was 15 cm3/m3 TWA (49 mg/m3). All biological measurements in this study gave the same assessment about exposure to benzene and correlated highly significantly with each other and with the air measurements (r = 0.8 or more). In the benzene factory the correlation was good even when calculated from samples with air concentration < 1 cm3/m3 (3.2 mg/m3) in the case of blood benzene and urinary MA. However, for S-PMA it was weak (r = 0.4) and for benzene in urine and exhaled air it did not exist any more. In the cokery, with mixed exposure, the correlation at low levels was weaker even for blood benzene and urinary MA (r = 0.6). According to the results in the benzene factory the exposure to pure benzene at the level 1 cm3/m3 (3.25 mg/m3) TWA gave: the blood benzene value about 110 nmol/l (8.6 micrograms/l), MA 23 mumol/l (3.3 micrograms/l) or 2.0 mg/g creatinine, S-PMA 58 micrograms/g creatinine or 0.4 mumol/l (95.7 micrograms/l), benzene in urine 499 nmol/l (39 micrograms/l), and benzene in the exhaled air 2.8 nmol/l (0.2 microgram/l). In general, the measurement of benzene in blood and in exhaled air, as well as benzene and its metabolites MA and S-PMA in urine, all gave similar results. However, at low exposure level (< 1 cm3/m3) the most reliable analyses were MA in urine and benzene in blood. PMID- 9200850 TI - A novel device for capturing breath samples for solvent analysis. AB - We have developed a novel breath sampling device suitable for capturing a portion of end-tidal air. This breath sample is then transferred onto a Perkin Elmer automated thermal desorption (ATD) sampling tube which is subsequently analysed by ATD-gas chromatography-mass spectrometry (GCMS). The breath sampler has been evaluated in the laboratory, in brief field trials and in human volunteer studies. The method is sensitive with a typical detection limit of 1 nmol/l and reproducible with an overall coefficient of variation between 5% and 15% for collection and analysis of breath samples from volunteers. The field trials used the sampler to assess exposure to solvents in several industries including the shoe manufacturing industry, the inks and coatings industry and at dry cleaning establishments. The sampler was found easy to use and reliable. Solvents detected include ethyl acetate (6.4-25.5 nmol/l), propan-2-ol (3.4-39.3 nmol/l), 2 butanone (0-6.6 nmol/l) and tetrachloroethene (0-557 nmol/l). The breath sampler was also used to monitor the elimination of solvents in breath from human volunteers after exposure chamber studies. More than 500 breath samples have been analysed from 24 volunteers in exposures to 10 different solvents (toluene, trimethyl benzene, tetrachloroethene, tetrahydrofuran, acetone, propan-2-ol, xylene, 2-butanone, 1-methoxy-2-propanol and n-hexane). The breath sampler allowed the rapid and non-invasive collection of data on elimination of solvents. PMID- 9200849 TI - Biological monitoring of experimental human exposure to trimethylbenzene. AB - Trimethylbenzene (TMB) is a component of numerous commercial preparations of organic solvents (Farbasol, Solvesso, Shellsol) used in the chemical, plastics, printing and other industries. TMB is a mixture of three isomers (pseudocumene 1,2,4-TMB; mesitylene-1,3,5-TMB; hemimellitene-1,2,3-TMB). The proportion of individual isomers in the mixture differs. The aim of this study was to obtain toxicokinetic data on the absorption and elimination of trimethylbenzene and its metabolites in biological fluids and to investigate the relationship between the biological indices of exposure and the absorbed dose. Eight-hour inhalation tests were performed in a toxicological chamber, The subjects were eight volunteers aged 20-39 with no history of exposure to TMB. They were exposed to pseudocumene, mesitylene or hemimellitene at concentrations ranging from 5 to 150 mg/m3 air. Exhaled air, capillary blood and urine samples were collected before, during and after the exposure. The determinations of TMB or its metabolites were performed using gas chromatography (HP 5890 II Plus, MSD, FID). Pulmonary ventilation in the volunteers ranged from 0.56 to 1.0 m3/h. The retention of 1,2,4-TMB; 1,3,5 TMB; 1,2,3-TMB in the lungs was 68%, 67% and 71%, respectively. The elimination of TMB from capillary blood occurred in accordance with the open three compartment model. Urinary excretion of dimethylbenzoic acids (DMBA) proceeded according to the open two-compartment model. Based on the toxicokinetic data, a simulation model of accretion and excretion of DMBA in urine during a 14-day period was developed. The highest rates of metabolite excretion and the highest quantities of DMBA in urine during 24-h intervals were observed on day 5 of exposure. The relationship between the levels of TMB or DMBA in biological material and TMB air concentration or absorbed dose were determined. To select the urine fraction suitable for determining occupational TMB exposure, linear regression analysis was performed. The biological exposure limit (BEL) for TMB has been proposed, with the current maximum allowable concentration (MAC) value of 100 mg/m3 (Polish standard) baseline value. PMID- 9200851 TI - Occupational exposure to chromium and nickel in the 1980s in Finland. AB - Two large data bases accumulated from the 1980s at the Finnish Institute of Occupational Health, one with results on urinary chromium and nickel analyses and the other with results on total and hexavalent chromium and nickel, were compiled and analysed in order to clarify the occupational exposure during the 1980s, and to reveal possible trends in the exposure of workers in different jobs. The data were processed in three batches: years 1980-1982, 1983-1985 and 1986-1989. The median values of urinary chromium exceeded the biomonitoring action level, BAL, in metal workers, and the mean values exceeded the BAL in both metal workers and plasma cutters. Among all worker groups the median values of urinary chromium remained quite stable during the study period. The median values of urinary nickel concentration did not exceed the BAL in any worker group studied, but an increasing trend was observed among moulders. In the breathing zone of grinders, the median value of total or trivalent chromium exceeded the occupational exposure limit, OEL. The medial of hexavalent chromium concentration in the breathing zone of metal sprayers and spray painters was higher than the OEL. No decreasing trend in exposure could be observed during the 10-year period in breathing zone air. PMID- 9200852 TI - Analysis of aluminium in serum and urine for the biomonitoring of occupational exposure. AB - A reliable and sensitive graphite furnace atomic absorption spectrometry (GFAAS) method with Zeeman background correction was developed for the analysis of aluminium in serum and urine in the biological monitoring of aluminium exposure. The method is based on platform atomisation in pyrolytically coated graphite tubes after fourfold dilution with nitric acid. For serum analysis, a matrix matched standard curve is prepared and for urine the method of standard additions is used. The within-run imprecision (C.V.) for serum and urine was 3% and 5%, and the between-day imprecision, 6% and 7.2%, at a concentration level of 4.0 mumol/l. The between-day imprecision for urinary aluminium was 15.7% at a concentration level of 0.24 mumol/l. The detection limits were 0.02 mumol/l for serum and 0.07 mumol/l for urine. During 1 year of participation in TEQAS external quality assessment scheme of the Robens Institute for Health and Safety (Guildford, UK) for serum aluminium the maximum cumulative performance score was achieved. For urinary aluminium a certificate in the external quality control scheme of the German Society of Occupational Medicine was obtained. The mean concentration of aluminium in a non-exposed population, who did not use antacid drugs, was 0.06 mumol/l (S.D. 0.03, range 0.02-0.13, n = 21) in serum, and 0.33 mumol/l (S.D. 0.18, range 0.07-0.82, n = 44) in urine. The upper reference limit for aluminium in a healthy, non-exposed population was estimated to be 0.1 mumol/l in serum and 0.6 mumol/l in urine. PMID- 9200853 TI - Biological monitoring of cadmium exposure and renal effects in a population group residing in a polluted area in China. AB - In an area of China, not previously studied in detail concerning cadmium pollution and possible adverse effects on the kidney of exposed populations, concentrations of cadmium in urine as an indicator of renal accumulation of cadmium was studied and related to indicators of renal dysfunction in order to examine if a relationship could be documented. Cadmium concentrations in urine were analysed by graphite furnace atomic absorption spectrometry and urinary beta 2 microglobulin (UBM) and albumin (UALB) were measured as indicators of renal dysfunction, Rice samples and urine samples were obtained from three areas in Zhejiang province, China, representing a highly exposed area, a medium exposed area and a control area, respectively. Cadmium concentrations in rice were 3.70, 0.51 and 0.072 mg/kg for the heavily, medium polluted areas and the control area, respectively. Cadmium concentrations in urine (geometric means) were 10.7, 1.62 and 0.40 micrograms/l in the high, medium and control areas respectively. There was a clear increase in UBM and UALB in the heavily exposed group in comparison to the control group and a slight increase in the medium exposed group. There was a statistically significant dose-response relationship between cadmium in urine and beta 2-microglobulin excretion in urine, which is similar to what has previously been reported in other countries. The findings constitute the first report concerning a dose-response relationship in this population group in Zhejiang province in China. PMID- 9200854 TI - An investigation of occupational metal exposure in thermal spraying processes. AB - A cross-sectional study of 34 workers engaged in thermal spraying at six worksites was undertaken in order to determine levels of exposure to and uptake of metals during different metal spraying activities. Levels of exposure to cobalt, chromium and nickel were highest in plasma sprayers and, on occasions exceeded UK Occupational Exposure Limits. Exposure to metals during detonation gun and electric arc spraying was better controlled and levels remained below the relevant Occupational Exposure Limits throughout the study period. Urinary levels of cobalt and nickel mirrored the airborne concentrations and the highest urine concentrations were again found in plasma sprayers. Urinary chromium levels were highest in electric arc sprayers, which may also reflect an increased body burden in this group due to a longer history of exposure. The findings clearly indicate that exposure to and uptake of metals may exceed UK Occupational Limits or Standards when spraying is performed manually or semi-automatically and where control relies on local exhaust ventilation (LEV) and personal respiratory protective equipment (RPE). PMID- 9200855 TI - Possible oral lead intake via contaminated facial skin. AB - Thirty-six workers exposed to low or moderate levels of lead at low temperature refining processes were surveyed to examine the route of lead intake. Blood lead level (BPb), delta-aminolevulinic acid in urine, lead in facial skin wipes (Face Pb) and lead in fingernails (Nail-Pb) were measured and their personal hygienic behavior was surveyed by a questionnaire. BPb showed a significant correlation with Face-Pb and Nail-Pb (r = 0.730 and r = 0.590, respectively). Multiple regression analysis extracted the factors of smoking at the workplace, face-Pb and nail-Pb as significantly related to BPb level. Electron-microscopic observation revealed that the majority of dust particles collected from worker's faces were larger than respirable size. Lead ingestion from contaminated face skin and fingers may contribute to elevations in the BPb level among workers. PMID- 9200856 TI - Estimation of individual dust exposure by magnetopneumography in stainless steel production. AB - The objectives of the study were to measure the magnetic dust lung burden of workers in stainless steel production by magnetopneumography (MPG) and to investigate the relationship of the results with air-borne concentrations of dust, total and hexavalent chromium as well as urinary excretion of chromium. There were 128 workers from the chromite mine, sintering plant, ferrochrome smelter, stainless steel smelting shop, cold rolling mill and welding shop in the exposed groups and five persons from the office staff in the control group. The remanent magnetic field (RMF) in the lungs was slightly elevated among workers in the ferrochromium and steel smelting shops; the levels were, however, lower than those reported for welders earlier and those observed in the welding/repair shop. Workers in the mine, concentrator and sintering plants and in the cold rolling mill exhibited remanent magnetic fields comparable to the referents. There was a relationship between the RMF and the actual urinary chromium concentration. Miners and concentrator and sintering plant workers showed retarded relaxation rate (ReR) of the remanent magnetic field. However, the RMF of the first two of these groups were low (< 0.1 nT) and this made it difficult to measure the ReR accurately. The duration of exposure correlated weakly but significantly with the relaxation rate, while smoking was not related to it. PMID- 9200857 TI - Methods for routine biological monitoring of carcinogenic PAH-mixtures. AB - The ability of a biomarker to provide an assessment of the integrated individual dose following uptake through multiple routes is especially valuable for mixtures of polycyclic aromatic hydrocarbons (PAH), due to methodological and practical difficulties of collecting and analysing samples from the various environmental compartments like air, water and soil and various media such as diet, cigarette smoke and workroom air. Since 1980, a large variety of novel approaches and techniques have been suggested and tested, e.g. urinary thioethers, mutagenicity in urine, levels of PAH or PAH-metabolites in blood and urine and methods for determination of adducts in DNA and proteins. Two approaches are more frequently reported: PAH-DNA-adduct monitoring in blood cells and urinary 1-hydroxypyrene monitoring. A large research effort has been made to use the extent of binding of PAH to DNA as a biomarker of exposure. The 32P-post-labeling assay detects the total of aromatic DNA-adducts and the adduct level in white blood cells is claimed to be an indicator of the biological effect of the PAH-mixture. However, the levels of aromatic DNA-adducts may be subject to appreciable analytical and biological variation. The present technical complexity of the method makes it more convenient for research applications than for routine application in occupational health practice. Pyrene is a dominant compound in the PAH mixture and is mainly metabolised to the intermediary 1-hydroxypyrene to form 1 hydroxypyrene-glucuronide, which is excreted in urine. Since the introduction of the determination of 1-hydroxypyrene in urine as a biomarker for human exposure assessment in 1985, many reports from different countries from Europe, Asia and America confirmed the potential of this novel approach. The conclusion of the first international workshop on 1-hydroxypyrene in 1993 was that urinary 1 hydroxypyrene is a solid biological exposure indicator of PAH. Studies with a comparison of several biomarkers confirmed that 1-hydroxypyrene in urine is a valid and sensitive indicator of exposure. Periodical monitoring of 1 hydroxypyrene appears to be a powerful method in controlling occupational PAH exposure in industries. The reference level and the biological exposure limit of 1-hydroxypyrene in urine are discussed. PMID- 9200859 TI - Urinary 1-hydroxypyrene levels of garbage collectors with low-level exposure to polycyclic aromatic hydrocarbons. AB - Because garbage collectors work in the street, they are exposed to polycyclic aromatic hydrocarbons (PAHs) in motor vehicle exhaust gas as they work. Urinary 1 hydroxypyrene (1-OH-pyrene) began to be used as a biological monitoring index for human exposure to high concentrations of PAHs. The objective of this study was to examine the applicability of urinary 1-OH-pyrene as a biological monitoring index for human low-level PAH exposure, such as the PAH exposure experienced while working in the street. The subjects were fifteen male garbage collectors. We measured individual exposure to PAHs, urinary 1-OH-pyrene concentrations and urinary cotinine concentrations. Individual air samplers were attached to the collar of the clothing of five workers to capture PAHs. Urine samples were collected before work, around noon and after finishing the day's work. In all, five PAH samples and 45 urine samples were collected. As control data, we analyzed the urinary 1-OH-pyrene and urinary cotinine levels of six smoking and four non-smoking control subjects who were not occupationally exposed to PAHs. The benzo[a]pyrene level in the air sampled for 5-6 h was 2.5-10.5 ng/m3, and the pyrene level as 10.3-70.3 ng/m3. These levels were similar to those in the vicinity of streets in Japan. A positive correlation between total PAH levels and the pyrene levels was observed. The average urinary 1-OH-pyrene level of the smokers was 0.21 +/- 0.13 mumol/mol creatinine, vs. 0.15 +/- 0.11 mumol/mol creatinine in the non-smokers. The urinary 1-OH-pyrene level obtained in this study was slightly higher than in the control group. No correlation was found between pyrene exposure and the urinary 1-OH-pyrene level of the five workers who wore the personal samplers. A significant positive correlation was observed between the urinary 1-OH-pyrene level and urinary cotinine level of the smokers. A significant positive correlation was also observed between the urinary 1-OH pyrene and urinary cotinine levels of the control group smokers. In conclusion, urinary 1-OH-pyrene is not applicable for biological monitoring of extremely low levels of exposure to PAHs, as in the case of working in the street. Caution is required to exclude the effects of smoking when evaluating PAH exposure. PMID- 9200858 TI - Ambient and biological monitoring of exposure to polycyclic aromatic hydrocarbons at a coking plant. AB - The exposure to polycyclic aromatic hydrocarbons (PAH) was measured in a Finnish coking plant over a 7-year period (1988-1994), since the beginning of production. Hygienic measurements including dust and vapour sampling were performed and the correlations between the concentrations of airborne pyrene with the levels of pyrene metabolite 1-pyrenol in urine were calculated. The profile of measured 12 or 15 PAHs was very similar between mean concentrations of personal samples, which suggests that it is possible to calculate the concentrations of total PAH by using e.g. pyrene as a marker compound. Measurements suggest that the progress of working conditions has been very favourable because the mean exposure level of shift workers to benzo[a]pyrene has decreased from 2.5 micrograms/m3 to 0.3 micrograms/m3. This points to successful measures of technical prevention. The mean concentration of 1-pyrenol in urine has been 0.2-0.6 mumol/mol creatinine. The concentration increases slightly towards the end of the working day, but the correlation urinary pyrenol and air pyrene was weak. Therefore the usefulness of pyrenol level for predicting the pyrene concentration at low exposure level in the ambient air is very limited. PMID- 9200860 TI - The influence of skin moisture on the dermal absorption of propoxur in human volunteers: a consideration for biological monitoring practices. AB - A large number of workers in agriculture are exposed daily (through skin contact) to pesticides either directly during mixing and loading or indirectly due to contact. The aim of this study was to investigate the influence of skin moisture on the dermal uptake of the pesticide propoxur. The study was conducted in human volunteers under controlled temperature conditions (30 degrees C) and environmental relative humidities of either 50, 70 or 90%. The study was approved by the Medical Ethics Committee. In this study a linear relationship between the environmental relative humidity and the level of skin moisture was observed. The results indicate that the level of skin moisture influences the absorption of propoxur via the dermal route, dramatically ranging from, on average, 13, 33-63% of the potentially absorbed dose' which is excreted in urine as the primary metabolite 2-isopropoxyphenol (IPP) at relative humidity levels of, on average 50, 70 and 90%, respectively. The 'potentially absorbed dose' is defined as the difference between the applied dose and the dislodged dose after 4 h. It can be concluded that by assessing health risks of workers in agriculture exposed dermally to pesticides and e.g. in testing the efficiency of protective clothing under realistic conditions, the influence of the level of skin moisture on absorption of substances may be considerable and has to be taken into account. PMID- 9200861 TI - Biological monitoring of pyrethroids in blood and pyrethroid metabolites in urine: applications and limitations. AB - The objective of this study was to perform biological monitoring of subjects who are occupationally exposed to pyrethroids. The study group consisted of 30 pest control operators exposed to cyfluthrin, cypermethrin or permethrin. After exposure, 24-h urine samples were collected and 20 ml of blood was drawn. The pyrethroid metabolites cis- and trans-3-(2,2-dichlorovinyl)-2,2 dimethylcyclopropanecarboxylic acid, 3-phenoxybenzoic acid and fluorophenoxybenzoic acid were determined in the urine samples (limit of detection: 0.5 micrograms/l) by GC MS and the pyrethroids in plasma (limit of detection: 5 micrograms) GC-ECD. The concentrations of metabolites in the urine of the pest control operators ranged between < 0.5 micrograms/l and 277 micrograms/l urine. The concentrations of cyfluthrin, cypermethrin and permethrin in the plasma were below the limits of detection (< 5 micrograms/l). To test if the metabolites are specific for pyrethroid exposure, they were determined in the urine of non-exposed subjects (n = 40). In no case could pyrethroid metabolites be detected. A cyfluthrin elimination experiment showed that cyfluthrin metabolites are eliminated following first-order kinetics (t 1/2 = 6.4 h). Storage experiments demonstrate that frozen urine samples (-21 degrees C) show no significant losses of metabolites within a year. In contrast, pyrethroids stored in plasma are susceptible to further biodegeneration. PMID- 9200862 TI - Assessment of environmental pollution by PCBs, DDT and its metabolites using human milk of mothers in Zimbabwe. AB - The milk samples were collected from mothers who had lived in the area for at least 5 years, healthy and breast feeding their first, second or third child. Of the 175 mothers' milk samples analysed, the organochlorine pesticide residues were detected in the following order of frequency: pp-DDE, 100%, pp-DDT 98%; and sum PCB, 53%. Of all the seven areas analysed the Kariba area and the highest mean level of sum DDT--25,259 ng/g milk fat and the lowest mean level of sum DDT of 1607 ng/G milk fat was found in Esigodini which is a rural area. The major DDT metabolite was pp-DDE. The ratio of pp-DDT/pp-DDE was highest in Kariba (0.6) suggesting recent pollution by DDT in that area. The results show that the vector control programmes (extensive pesticide spraying of disease-carrying pests, such as mosquitoes and tsetse flies), agricultural activities and dietary habits were the main contributing factors towards the high levels of pesticides in most of the areas. Kadoma area had the highest mean level of sum-PCB (60 ng/g milk fat). PMID- 9200863 TI - Recent policy and technical developments in biological monitoring in the United Kingdom. AB - In 1996 the United Kingdom's Health and Safety Executive introduced biological monitoring guidance values for six substances, butoxyethanol, N,N dimethylacetamide, lindane, methylene-bis(2-chloroaniline), mercury and methylenedianiline. These guidance values were set as either health-based values or hygiene-based values calculate according to the 90th percentile (benchmark concept). Recent technical developments from the Health and Safety Laboratory are described in this paper and include: (i) the use of breath analysis as a useful non-invasive routine monitoring technique; (ii) flow cytometry as a means for measuring different patterns of immune cell activation from workers exposed to respiratory sensitisers when compared with those exposed to chemical irritants; (iii) the use of molecular techniques to explore the possible role of individual susceptibility in the development and severity of glomerulonephritis; (iv) the development of expert systems for predicting the skin permeability of chemicals, and respiratory and skin sensitisation. PMID- 9200864 TI - Large-scale biological monitoring in Japan. AB - Data from the large-scale biological monitoring program in Japan were assembled and analyzed and the following results were obtained. All workers handling lead and eight kinds of major organic solvents received physical examinations and biological monitoring at the same time. Therefore, the number of workers handling industrial chemicals and that received physical examinations and the number of workers been examined by biological monitorings were similar to each other. The total number of cases examined from 1989 to 1994 was about 661,000 for lead in the blood and about 4,173,000 for the urinary metabolites of eight organic solvents. The results were classified into three categories and category 3 consists of workers having exposure concentrations above the 1988-1989 biological exposure indices of the ACGIH with the exception of lead concentration in the blood where the limit in Japan was set at 40 micrograms/100 ml. The percentage of exposed workers in category 3 was 1.4% for blood lead and 0.2-2.4% for the urinary metabolites of the eight organic solvents. The percentage of exposed workers in category 3 for blood lead, delta-aminolevulinic acid, urinary mandelic acid, N-methylformamide and 2,5-hexanedione in the urine has decreased with time. In ambient monitoring, the percentage of workplaces in classification 3 for lead and styrene also has decreased with time. PMID- 9200865 TI - Biomonitoring of early effects on the kidney or the lung. AB - Biomarkers of toxicity are usually altered before the onset of functional changes or clinical manifestations. Peripheral biomarkers of toxicity present an additional advantage in that they can be applied on easily accessible biological materials (blood or urine). They represent useful tools that can be used for identifying subjects or groups at risk in the industry or environment, or for establishing acceptable exposure levels. In the case of the kidney, for instance, a relatively large battery of peripheral markers has been developed during the last decade. These markers are either constituents of the renal parenchyma whose urinary excretion can signal lesions or an abnormal secretion, or plasma proteins reflecting the integrity of nephron structures involved in their selective filtration or reabsorption. The epidemiological application of these tests has revealed that widespread occupational or environmental pollutants such as lead, cadmium, crystalline silica or perchloroethylene may cause very early effects on different segments of the nephron. In the case of the lung, we have recently identified a lung toxicity biomarker applicable not only on bronchoalveolar lavage fluid but also or sputum provided some precautions are taken. This biomarker called Clara cell protein (CC16) is a kDa protein secreted in the respiratory tract by the non-ciliated Clara cells known for their vulnerability to toxic insult. Studies on occupationally exposed workers and experimental animals indicate that the assay of CC16 in serum is a sensitive and a relatively specific test to detect early acute or chronic effects of toxicants on the tracheobronchial tree. The ultimate goal, however, is the development and validation of biomarkers that have a sufficient toxicological relevance to be used for health risk assessment. It is thus of premier importance to establish in health significance of early biomarkers of toxicity by determining to what extent they reflect critical steps in, and are predictive of the development of a chronic and irreversible disease. PMID- 9200866 TI - A new concept for risk assessment of the hazards of non-genotoxic chemicals- electronmicroscopic studies of the cell surface. Evidence for the action of lipophilic chemicals on the Ca2+ signaling system. AB - Recently, we presented evidence for the localization of components of the cellular Ca2+ signaling pathway in microvilli. On stimulation of this pathway, microvilli undergo characteristic morphological changes which can be detected by scanning electron microscopy (SEM) of the cell surface. Here we show that both receptor-mediated (vasopressin) and unspecific stimulation of the Ca2+ signaling system by the lipophilic tumor promoters thapsigargin (TG) and phorbolmyristateacetate (PMA) are accompanied by the same type of morphological changes of the cell surface. Since stimulated cell proliferation accelerates tumor development and sustained elevation of the intracellular Ca2+ concentrations is a precondition for stimulated cell proliferation, activated Ca2+ signaling is one possible mechanism of non-genomic tumor promotion. Using isolated rat hepatocytes we show that all tested lipophilic chemicals with known tumor promoter action, caused characteristic microvillar shape changes. On the other hand, lipophilic solvents that were used as differentiating agents in cell cultures such as dimethylsulfoxide (DMSO) and dimethylformamide also, failed to change the microvillar shapes. Instead DMSO stabilized the original appearance of microvilli. The used technique provides a convenient method for the evaluation of non-genomic carcinogenicity of chemicals prior to their industrial application. PMID- 9200867 TI - Urinary 1-hydroxypyrene as a marker of exposure to pyrene: an epidemiological survey on a general population group. AB - Urinary levels of 1-hydroxypyrene in a general adult population group are studied. Experimental data are not normally distributed; statistical analysis required a base 10 logarithmic transformation of data. The concentrations of urinary 1-hydroxypyrene measured were expressed as microgram g-1 urinary creatinine and are comparable with those reported by other authors, both for smoker and non-smoker subgroups. Multiple regression analysis shows that, for smokers, the number of cigarettes smoked per day and the body mass index (BMI) significantly influence the levels of urinary 1-hydroxypyrene expressed as microgram g-1 urinary creatinine, whereas no personal or behavioural variable (age, sex, alcohol consumption, dietary intake of pyrene, BMI) modified the 1 hydroxypyrene levels for non-smokers. PMID- 9200868 TI - Trace metals in field samples of zooplankton from the Fram Strait and the Greenland Sea. AB - Trace metals (Cd, Pb, Ni, Cu, Zn and Hg) were evaluated in 14 zooplankton taxa collected on cruise ARK IX/Ib of RV 'Polarstern' to the Fram Strait and the Greenland Sea in March and April 1993. We found a substantial interspecific heterogeneity, e.g. with rather low Cd concentrations in calanoid copepods (0.1 0.7 mg kg-1, dry wt.) but remarkably high levels in the decapod Hymenodora glacialis (7-9 mg kg-1) and in the amphipods Themisto abyssorum and T. libellula (24-34 kg-1). In general, Pb was low (< 1 mg kg-1), while some enhanced Ni concentrations were found in the ostracod Conchoecia borealis (66-86 mg kg-1). A comparison to world-wide reported data on marine crustaceans did not reveal any suggestions on increased metal availabilities in both areas investigated, although one might expect a transport of some metals from Siberian rivers across the pole by the Transpolar Ice Drift Stream. However, more information on accumulation strategies of zooplankton under winter and summer conditions is necessary before a full assessment of metals in Arctic waters will be possible. PMID- 9200870 TI - Different element ratios of red cosmetics excavated from ancient burials of Japan. AB - Marker elements of red cosmetics, collected from ancient burials of Matsuyama, Tokushima and Nara Japan, were determined by emission spectrometry (ICP/AES). The mass ratios of Hg, Fe, Cu, and Zn were different between samples. Element levels were compared with reference to relative amounts of sulfur. Of the possible contaminants from the bone and sand of burials, the relative amounts of Hg and Fe to S were most commonly available to evaluate the difference between the cosmetics. The cosmetics were divided into four groups; type I (high Hg with less Fe), type II (both moderate Hg and Fe), type III (moderate Hg with high Fe) and type IV (less Hg with high Fe). The main constituents of cosmetics are mercury sulfide (cinnabar) or ferric oxide mixed with trace metals. Zinc contents differ between the Fe and Hg amounts for the three areas. Cosmetic compositions varied with each burial site, suggesting that they were derived from different mines of ancient Japan. PMID- 9200869 TI - Studies on leaching of Cr and Ni from stainless steel utensils in certain acids and in some Indian drinks. AB - Leachates of Cr and Ni from stainless steel utensils viz., frying pans, bowls and tumblers, have been investigated, by exposing the utensils to decinormal solutions of citric, tartaric and lactic acids and to some common Indian drinks. A comparison of observed results indicate that the complexation of metal ions with organic acid anions is most vital and metal leaching is largely a function of the availability of free anions. The intake of Cr and Ni by human beings has also been calculated. PMID- 9200871 TI - An international symposium on neonatal hypoglycemia: thirty years later. Does it injure the neonatal brain? AB - A discussion of neonatal hypoglycemia was held on 18 November 1995 as a satellite symposium of the 40th Annual Meeting of Japan Society for Premature and Newborn Medicine and continued in closed session of 19 November to address neonatal hypoglycemia in the 21st century. This represented a 30-year follow-up of a discussion of carbohydrate and energy metabolism in the newborn held in Tokyo on 10 November 1965. This follow-up was prompted by the incredible advances in clinical care in perinatal medicine and in basic knowledge in the neurosciences, neonatal physiology and metabolism that have occurred in Japan and around the world throughout these 3 decades. PMID- 9200872 TI - Neonatal hypoglycemia 30 years later: does it injure the brain? Historical summary and present challenges. AB - Since 1911, blood sugars have been measured in newborn infants. Significant neonatal hypoglycemia was first reported in 1937. In 1959, the report of transient symptomatic neonatal hypoglycemia generated worldwide reports. This, along with the ongoing advances in studies of energy metabolism, thermal control and oxygen requirements, led to the first conference on Energy and Carbohydrate Metabolism in the newborn in Tokyo, 1965. Subsequently, a number of hypoglycemia syndromes were discovered. Concurrently, pre-, peri- and neonatal care changed dramatically with the survival or very tiny and very sick newborns. These advances in care made previously derived statistical definitions of hypoglycemia irrelevant. New functional definitions are needed to define abnormal glucose concentrations. Significant hypoglycemia is a continuum of low glucose concentrations of varied duration and severity. Its impact depends upon other risk factors as well. In addition, new hypoglycemic syndromes have appeared. These include deficiencies of blood-brain glucose transporters, the association of hyperinsulinemic hypoglycemia with isoimmune thrombocytopenia and a variety of acyl CoA dehydrogenase deficiencies. Concurrently, carbohydrate disorders in infancy appear to be changing. Neonatal diabetes mellitus, previously transient and benign, now shows a high frequency of recurrence and remaining as a permanent condition. Idiopathic ketotic hypoglycemia of infancy has disappeared in the USA. Familial hyperinsulinemic hypoglycemic syndromes of infancy appear to have a good prognosis, respond to medical intervention and have had their genetic defect localized to a specific gene. Current advances promise reliable bedside techniques to measure central nervous system function, cerebral blood flow, endocrine hormones and receptors as well as glucose transporters and specific genetic defects. These data, when correlated with plasma glucose concentrations and central nervous system function and development, should provide a better understanding of the impact of prolonged and profound hypoglycemia on long-term outcome. PMID- 9200873 TI - Hypoglycemia in the neonate: current controversies. AB - An overview of current controversies in the field of neonatal hypoglycemia is presented. Such controversies include the biochemical definition of hypoglycemia, the methods of monitoring blood glucose concentrations at the bedside, and the effects of hypoglycemia on the brain, together with the relationship between neonatal symptoms, metabolic status and treatment, and neurological outcome. The historical background to this confusion is reviewed, emphasizing that much of this contention arises from analyses of blood glucose concentrations in the first postnatal days, performed on neonates over 30 years ago in which no consideration of the effects of medical management and nutritional policy was taken into account in interpreting the subsequent blood glucose profiles. It is emphasized that there is no evidence to support the perception that the brain of the low birthweight infant is more tolerant to low blood glucose concentrations that that of the fullterm infant. Possible approaches to definition are presented from which it is concluded that the definition of hypoglycemia should be: the lowest concentration of glucose which in combination with other metabolic fuels allows normal brain function. It is emphasized that hypoglycemia is a continuum, no single blood glucose concentration reflecting functional changes in every infant at that level. Confusion over the diagnosis of hypoglycemia is compounded by the inadequacy of currently available methods of bedside monitoring for blood glucose concentrations in the newborn nursery. A pragmatic approach to monitoring and to treatment is presented, suggesting that any infant at risk with a blood glucose concentration less than 2.6 mmol/L should be monitored thereafter, especially in relation to blood glucose changes before and after feeding. Finally, it is emphasized that the study that needs to be done is one which there is documentation of glucose and other fuel concentrations in relation to neurophysiological function during the newborn period and linked with randomized controlled trials of interventions, with follow-up into later childhood. PMID- 9200874 TI - Approaches to the definition of neonatal hypoglycemia. AB - There is no current agreement on the definition of neonatal hypoglycemia. Three different bases for the definition are presented, with a review of evidence of relevant to each approach. (1) Not commonly encountered; requires evidence of normal distribution of blood glucose concentrations. (2) Increased risk of adverse sequelae; requires evidence of short-term and long-term sequelae of different blood glucose concentrations. (3) Benefits of intervention outweigh risks; requires evidence from randomized trials of intervention. The usual distribution of neonatal blood glucose concentrations varies with birthweight, postnatal age, nutritional intake and other factors. Clinical signs occur in some but not all babies with low blood glucose concentrations, but such signs lack specificity. Abnormality in sensory evoked potentials is associated with variations in blood glucose concentrations; these changes suggest that values less than 2.6 mmol/L have acute effects in areas of the brain with high glucose demand. Among low birthweight babies, case series suggest a high risk of impairment (50%) following symptomatic hypoglycemia, and controlled studies generally confirm an association between very low neonatal blood glucose concentrations and adverse neurodevelopment. However, these studies do not establish a level of blood glucose concentration or duration of hypoglycemia that is critical. There are no reports of randomized trials that have assessed the effect on neurodevelopment of alternative policies of glucose provision in the neonatal period. There is a need for a randomized controlled trial to assess reliably the short- and long-term clinical effects of alternative policies of the clinical management of blood glucose concentration in babies at high risk both of low neonatal blood glucose concentrations and adverse neurodevelopment. PMID- 9200875 TI - Neonatal insulin secretion: implications for the programming of metabolic homeostasis. AB - Patterns of metabolic adaptation are described in the neonate, which generate two fundamental concepts. First, that early nutritional experiences may have long term effects on the control of metabolic homeostasis, and second, that insulin has a fundamental role in this process. The endocrine pancreas in the neonate is unable to regulate insulin secretion in relation to blood glucose concentration with the same level of tight control seen in the older child and adult. Moreover, the pattern of metabolic adaptation in the fullterm infant in the first postnatal week is different to that of the preterm baby and the infant born small-for gestational-age (SGA), with both preterm and SGA infants being unable to generate counter-regulatory ketogenesis as blood glucose concentrations fall. The inability to initiate ketogenesis and switch off insulin secretion after birth persists for several weeks in preterm infants. Methods of feeding term and preterm infants have profound effects on the neonatal endocrine milieu and it is suggested that patterns of insulin secretion provoked in the newborn period may 'programme' the subsequent development of metabolic control. The recently described molecular mechanisms that underlie the pathogenesis of abnormal insulin secretion in the syndrome of persistent hyperinsulinaemic hypoglycemia of infancy (or pancreatic nesidioblastosis) may offer insights into how such programming may occur. PMID- 9200876 TI - Cerebral metabolic consequences of neonatal pathologies in the immature rat. AB - The cerebral metabolic consequences of hypoxia, seizures and hyperbilirubinemia were explored in immature rates between the postnatal age of 10 (P10) and 21 days (P21) by the quantitative autoradiographic [14C]2-deoxyglucose technique. The effects of a previous bilirubin exposure on cerebral regional permeability to bilirubin were measured by autoradiography. Hypoxia was induced by breathing a 7% N2/93% O2 gas mixture and seizures were initiated by injections of pentylenetetrazol. Hyperbilirubinemia was induced by the perfusion of a bilirubin/albumin solution. Hypoxia and seizures induced a general increase in cerebral metabolic rates to glucose (LCMRglc) in P10 rats, except in hippocampus during seizures. At P14, LCMRglc remained increased during seizures, except in the hippocampus. During hypoxia LCMRglc were unchanged in the genu of the corpus callosum and the anterior commissure and decreased in the cerebellar white matter. At P21, LCMRglc decreased in all white matter regions during hypoxia and in the hippocampus during seizures, while they were unchanged in the amygdala and increased in the nucleus of the solitary tract. During hyperbilirubinemia, LMCRglc decreased at all ages with very marked changes in the nucleus of the auditory nerve at P10 and in the inferior colliculus at P21 (72-86%). Twofold decreases were also recorded in the hippocampus. The basic regional cerebral permeability to the anion was higher at P10 than P21 and the marked increases in regional permeability to bilirubin after a previous exposure to the anion were located in the nucleus of the auditory nerve and the hippocampus. PMID- 9200877 TI - The value of neurophysiologic approaches in the anticipation and evaluation of neonatal hypoglycemia. AB - The association of low blood glucose with central nervous system (CNS) injury was first described in 1937 by Hartmann and Jaudon. In the early 60 years since publication of these observations the effects of hypoglycemia upon the brain remain poorly understood. Technology capable of accurately determining plasma glucose concentrations has been developed. Investigators have sought to establish critical values below which glucose levels should not be allowed to fall. Despite these efforts the definitive level of glucose capable of producing brain injury in any particular patient remains unknown. Glucose homeostasis within the neonatal CNS represents a dynamic process consisting of many interrelated variables including gestational and chronologic age, genotype, relative health, blood flow, metabolic rate and availability of other suitable substrates. New technique for assessing the glucose delivery: consumption ratio and directly monitoring the cellular consequences of glucose deprivation within discrete regions of the brain will help to answer the question 'How long is too low and how long is too long?' PMID- 9200878 TI - Hypoglycemia in healthy, full-term, breast-fed neonates during the early days of life: preliminary observation. AB - To examine the incidence of symptomatic and asymptomatic hypoglycemia during the early days of life, the blood glucose levels were analyzed in 38 healthy, full term, breast-fed neonates cared for by rooming-in immediately after birth. Blood glucose levels were measured randomly using a blood glucose analyzer from birth to discharge. Preliminary results have shown that hypoglycemia (< 40 mg/dL) seldom occurred in healthy, full-term, breast-fed neonates when cared for in rooming-in with frequent suckling immediately after birth. PMID- 9200879 TI - Neonatal hypoglycemia in infants with intrauterine growth retardation due to pregnancy-induced hypertension. AB - The early-onset type (onset earlier than 28 gestational weeks) of pregnancy induced hypertension (PIH) has the clinical characteristics of a high incidence of intrauterine growth retardations (IUGR), fetal distress, neonatal hypoglycemia and hypertensive disposition. Moreover, the infants from early-onset type of PIH mothers showed a statistically significant higher incidence of neurological handicap (cerebral palsy, mental retardation and epilepsy) than late onset type. The infants with a neurological handicap had severe IUGR and intractable hypoglycemia in the early neonatal period, probably due to low storage of glucose in the liver and fetal hypoxia. Appropriate perinatal management, including proper evaluation of fetal well-being and good timing of delivery, could improve the outcome of infants from early-onset type of PIH mothers. PMID- 9200880 TI - Follow-up study of neonatal hypoglycemia. AB - A follow-up study was done on the neurological handicap and intellectual development of high-risk infants with neonatal hypoglycemia. The frequency of neonatal hypoglycemia in high-risk infants is 8.6%, and the incidence of a major neurological handicap in high-risk infants with neonatal hypoglycemia is 11%. Risk factors for the handicap group are very low birthweight, intrauterine growth retardation, perinatal asphyxia and mothers with pregnancy-induced hypertension. Average DQ at 1.5 and 2.5 years and IQ at 4 and 6 years of very low birthweight infants with neonatal hypoglycemia showed no significant difference, in contrast to the control group. PMID- 9200881 TI - Pre-adolescent chumship as a buffer against psychopathology in adolescents with weak family support and weak parental bonding. AB - This study examines the degree to which the existence of a pre-adolescent "chum" interacts with family and social environments to buffer mental distress in adolescents. 831 high school students participated in this study, (male: 355; female: 476; mean age 16.7 +/- 1.0). Subjects were administered questionnaires assessing psychopathology and support systems. A pathway analyses model was used to investigate pathways and their interrelationships from chum to psychopathology and from social and family support to psychopathology. Only when adolescents experience weak parental bonding does chumship have a role in buffering distress. PMID- 9200882 TI - Children's use of transitional objects: parental attitudes and perceptions. AB - Parental attitudes and perceptions toward children's use of transitional object among 75 sets of parents were examined. Overall, parents appear to understand the significance and importance of children's attachment to transitional objects, thus providing consistency in childrearing. Parental differences emerged in the following areas: situations when children want their transitional object, age children should give up transitional object use, and age parents gave up their own transitional object. Some racial and socioeconomic differences were found, however these differences must be viewed within the context of a broader cultural perspective. The study suggests that parents are perceptive and, for the most part, respectful of the significance of transitional objects in the lives of children. PMID- 9200883 TI - Child psychiatric patients' interactions with their mothers. AB - This study investigated whether child psychiatric patients' and their mothers interacted differently as a function of whether the children were diagnosed as having internalizing or externalizing disorders. Twenty children and their mothers were rated on eight behavior dimensions as they engaged in a ten-minute play session. Maternal depression was found to interact with their children's diagnoses and behavioral ratings. PMID- 9200884 TI - The Fort Bragg continuum of care Demonstration Project: the population served was unique and the outcomes are questionable. AB - Examination of the evaluation sample and the outcome data from the Fort Bragg Demonstration Project suggests that the children served were mildly disturbed, were atypical of those served in most public mental health clinics, spent less than optimal time in the new services developed, and were judged as making considerable progress with minimal treatment regardless of age or level of judged psychopathology. The use of normative outcome measures in a pre-post design was considered a major reason for failure to find any significant differences between differently treated children. PMID- 9200885 TI - Trichotillomania and related disorders in children and adolescents. AB - Eleven chronic hair pullers, 11 subjects with obsessive-compulsive disorders (OCD), and 11 subjects with a non-OCD anxiety disorder were assessed with structured interviews and the Child Behavior Checklist (CBCL). Only 4 hair pullers (36%) reported both rising tension and relief with hair pulling. Each group had significantly more internalizing than externalizing symptoms on the CBCL. Seven hair pullers (64%) had a lifetime history of at least one other axis I diagnosis. The results provide further evidence that trichotillomania in referred children and adolescents is usually a chronic disorder often associated with internalizing symptoms and psychiatric comorbidity. Rising tension followed by relief with hair pulling may be an unnecessary restriction in the diagnosis of childhood trichotillomania. PMID- 9200886 TI - Standard values for reproductive and clinical chemistry parameters of Japanese quail. AB - Historical control data of Japanese quail collected from reproduction studies conducted at the Federal Institute for Health Protection of Consumers and Veterinary Medicine between 1988 and 1994 are presented in this paper. Reproductive and clinical chemistry data from control animals of 10 ecotoxicological studies are summarized and discussed to develop a normal data base of this species. The data obtained were compared to the control reproductive parameters of bob-white quail and mallard ducks available in the literature. For a long time these two species have been most used in avian reproductive toxicology studies. In summary, the data obtained indicate that Japanese quail appears to be more appropriate to be used for the determination of reproductive effects of pesticides on birds. PMID- 9200887 TI - Failure to induce formation of proteinase K resistant fibrils in pigeons through experimental infection with paramyxovirus type 1. AB - Ten racing pigeons were infected experimentally with the paramyxovirus (PMV) type 1 of the pigeon. Within twelve weeks of observation, they were euthanized at different times. Their brains were examined for proteinase K resistant fibrils and histopathologically for spongiform lesions. No proteinase K resistant fibrils and no spongiform lesions could be detected in any case. Therefore, it is estimated that PMV type 1 of the pigeon is not likely to induce pathogenic mechanisms assumed for transmissible spongiform encephalopathies. PMID- 9200888 TI - Tissue concentrations and pharmacokinetics of florfenicol in broiler chickens. AB - Florfenicol was once administered to broiler chickens via i.v., i.m. and oral route (30 mg/kg body weight) to study its plasma concentrations, kinetic behaviour, systemic bioavailability and tissue levels. Following a single i. v. injection, the kinetic disposition of florfenicol followed a two-compartmental open model with an elimination half-life of 172 min, total body clearance of 26.9 ml/kg/min and a steady state volume of distribution of 5.11 litre/kg. The highest plasma concentrations of florfenicol were 3.82 and 3.20 micrograms/ml following single i.m. and oral administration, respectively. The systemic bioavailability was 96.6 and 55.3 per cent after i.m. and oral administration. The plasma protein binding of florfenicol was 18.5%. Following the administration, the highest tissue concentration of the drug was found in kidney, bile, lung, muscle, intestine, heart, liver, spleen and serum. Low concentrations of the drug were found in brain, bone marrow and fat. No florfenicol residues were detected in tissues and serum after 72 h except in the bile, it disappeared after 96 h. PMID- 9200889 TI - Orthodontic treatment and temporomandibular disorder: is there a relationship? Part 2: Clinical implications. AB - Although a review of the current literature does not reveal compelling evidence for the claim that orthodontic treatment prevents, causes, or cures temporomandibular disorders (TMD), the currently available clinical studies devoted to this topic share some methodological weaknesses, some of which are discussed in the present article. Another purpose of this paper is to extend the current understanding about the relationship between orthodontics and TMD to situations occurring in routine clinical practice. By doing so, we provide suggestions that are intended to help the orthodontist in decision-making when he or she deals with a patient who is in need of orthodontic treatment for dental or skeletal reasons, but has a history of TMD, or who develops TMD signs and symptoms during or after treatment. PMID- 9200890 TI - Mesiodentes: incidence, morphology, etiology. AB - There are many publications in the literature focusing on clinical, radiological and surgical aspects of the treatment of mesiodentes. However, the etiology of this dental anomaly remains widely unclear. The purpose of this study was to evaluate etiologic factors for mesiodentes in a collective comprising 30 patients with a total of 45 mesiodentes. Thirty-one percent of the patients showed a familial disposition, pointing to inheritance as a key factor in the development of mesiodentes. Our results further support the hypothesis of related etiologic factors for several dental and craniofacial anomalies, such as hyperdontia, hypodontia and cleft lip and palate. Finally, we report the gemination of a deciduous incisor on the same side as a mesiodens. We also found differences in the mesiodistal width of central incisors depending on unilateral or bilateral occurrence of mesiodentes. Both these findings support the dichotomy theory of the split in the tooth bud inducing the development of mesiodentes, a theory we favor over that of local hyperactivity of the dental lamina. PMID- 9200891 TI - Influence of facial growth pattern on outcome of extraction therapy. AB - The present clinical-radiological study analyzes orthodontic casts and lateral cephalometric X-rays (at start and finish of orthodontic treatment) of 56 extraction cases, most of them adolescent patients who had 4 teeth extracted. The test group was classified into 3 morphological categories according to growth patterns. For comparison purposes, cephalometric findings of morphologically matching non-extraction groups as well as corresponding data from the literature were used. There was no deepening of overbite in any of the extraction cases in the different test groups. However, an average bite opening of 1.2 mm was found in patients with a neutral or horizontal growth pattern. Irrespective of the growth pattern, a significant increase in anterior and posterior facial height as well as a mean reduction of the ANB angle between 0.9 degrees and 1.3 degrees was found in the different extraction groups. These results matched those of the corresponding nonextraction control groups. At the end of treatment, the longitudinal axis of the upper incisors appeared too steep (retruded). Overall, the individual growth pattern was found to be of very little relevance to treatment results, provided a well considered treatment plan had been drawn up. PMID- 9200892 TI - Luscher colour test in orthodontic patients. Practicability for compliance assessment? AB - The Luscher colour test has been repeatedly recommended as a tool for the prediction of orthodontic treatment course and patient cooperation despite its scientific shortcomings. A series of studies involving patients aged 9 to 16 years undergoing treatment with removable appliances at various orthodontic practices and at one university clinic provided no evidence of systematic correlations between Luscher test colour preferences and cooperation with appliance wear. There was only slight consistency in colour preferences in a re test conducted a few months after baseline measurements. Statistical evaluation of specific hypotheses and exploratory analysis of the global potential of the test provided no support for generalizable and useful correlations between colour preferences and patient compliance. Therefore the Luscher test cannot be generally recommended for the rating of patient compliance in the clinical orthodontic practice. PMID- 9200894 TI - Viggo Andresen--a pioneer in orofacial orthopedics. PMID- 9200893 TI - Light-cured glass ionomer cement as a bracket adhesive with different types of enamel conditioners. AB - Eighty bovine incisors were ground on 320-grit silicone carbide paper and cleaned with fluoride-free prophylaxis paste. The enamel surface conditions were: 1. no conditioning; 2. salicylic acid (10%, 10s); 3. benzoic acid (10%, 10s); 4. air polishing with sodium hydrogen carbonate/Prophy-Jet; 5. Prophy-Jet, followed by polyacrylic acid (PAA, 10%, 10 s); 6. PAA, followed by saliva contamination; 7. PAA; 8. phosphoric acid (37%, 10 s). Fuji Ortho II LC (GC) was used as a bracket adhesive in groups 1 t0 7, and in group 8 Concise orthodontic (3M). Stainless steel lingual buttons were placed by hand. Polymerisation with visible light was carried out 20 s from mesial, distal, incisal and gingival. After 24 h storage in tap water at room temperature the shear bond strengths were tested in accordance with ISO specification TC 106/SC/WG16. Mean values of the groups were compared using Student's t-test. Group 7 (PAA) attained the highest mean shear strength (in comparison with control group): 28 MPa. This was both significantly different from the control group (Concise, 33 MPa) and highly significant in comparison with the other groups (< 16 MPa). The shear bond strength of Fuji Ortho II LC on PAA conditioned enamel indicates the clinical applicability of this material. PMID- 9200896 TI - The naming of antibiotics. PMID- 9200895 TI - Grave violations. PMID- 9200897 TI - The diagnosis and treatment of post-stroke depression. AB - Depression is common after stroke but the diagnosis may be difficult. Anterior and subcortical brain lesions increase the risk of depression. It is important to make an accurate diagnosis of PSD as treatment may reduce morbidity. Treatment should consist of family support, education and anti-depressant medication. PMID- 9200898 TI - Stroke rehabilitation. PMID- 9200899 TI - Obesity hypoventilation [corrected] syndrome in the differential diagnosis of a pulmonary mass. AB - Dyspnea and cyanosis are common presenting manifestations of cardiopulmonary disease. When these findings occur in a cigarette smoker with an apparent pulmonary mass on chest radiograph, the differential diagnosis rapidly narrows to a short list of possibilities that include pulmonary neoplasm, pulmonary infection and pulmonary infarction. Pulmonary hypertension with pulmonary arterial enlargement and hypoxia secondary to alveolar hypoventilation should also, however, be included as a diagnostic possibility in the appropriate setting because the evaluation and treatment of this entity may differ markedly. PMID- 9200900 TI - Pulmonary actinomycosis: a cause of adult respiratory distress syndrome. PMID- 9200901 TI - Managed care--complaints and regulations. PMID- 9200902 TI - Proposed breast cancer screening recommendations. PMID- 9200903 TI - Percussion and stamps. PMID- 9200904 TI - The challenges of managed care. PMID- 9200905 TI - The psychiatric unit comes to the general hospital: a history of the movement. PMID- 9200907 TI - Medication noncompliance. PMID- 9200906 TI - Toward an integrated approach to the study of inpatient treatment of schizophrenia. PMID- 9200908 TI - Family involvement in general hospital inpatient care. PMID- 9200909 TI - Treatment of comorbid schizophrenia and substance abuse disorders. PMID- 9200910 TI - Discharge planning in psychiatric units in general hospitals. PMID- 9200911 TI - Meaningful linkage practices: challenges and opportunities. PMID- 9200912 TI - The future of inpatient psychiatry in general hospitals. PMID- 9200913 TI - [Molecular immunology on autoimmune disease]. PMID- 9200914 TI - [Clonal deletion and clonal anergy as the mechanism of self-tolerance induction]. AB - In the immune system, it is most important to discriminate self from nonself and to acquire and maintain the unresponsiveness to self antigens, self-tolerance. Recently, the transgenic animals provides the evidence of the mechanism of self tolerance induction. Self-tolerance is established mainly by elimination, clonal deletion, and functional inactivation, clonal energy, of the autoreactive T cells and B cells. Clonal deletion and clonal anergy are involved not only in the central tolerance, in thymus or bone marrow, but also in the peripheral tolerance. The failure of self-tolerance involves the induction of auto-immune disease. PMID- 9200915 TI - [Molecular mechanism of apoptosis operated in the immune system]. AB - Apoptosis is the physiological cell death, and plays an important role in the various fields of immunology, especially (1) in the formation of T cell and B cell repertoires, (2) in the regulation of hyper-activated T and B cells by Fas FasL, and (3) in the effector phases of eliminating potentially dangerous cells. Although, the exact mechanism of cell death is still unknown, progress has been made in identifying key elements in apoptosis. Particularly, ICE family was found essential for apoptotic execution, and Bcl-2 related proteins can block the molecular events of apoptosis. This article briefly reviewed such "on" and "off" signals in apoptosis, and discussed from the view point that the disturbance of the balance of these signals are relevant to the etiology of autoimmune diseases. PMID- 9200916 TI - [T cell tolerance and autoimmune disease]. AB - T cell tolerance is ensured by several complementary mechanisms arranged in a fail-safe manner, which contain clonal deletion in the thymus, and clonal anergy, activation-induced cell death, and suppression in the periphery. What is still unclear is the conditions under which, and the mechanisms by which, all these T cell tolerance systems can break down, inducing autoimmune phenomenon and then autoimmune disease. Fas or FasL mutant and CTLA-4 knock-out mice have successfully unraveled the molecular mechanisms for breakdown and maintenance of some T cell tolerance. Although they are of great theoretical interest, their involvement in the pathogenesis of the major autoimmune diseases remains controversial. In this minireview discussed are their precise mechanisms and possible importance in therapeutic immune intervention. PMID- 9200917 TI - [Mechanisms for B cell tolerance and their defects in systemic autoimmune diseases]. AB - Lines of evidence suggest that self-reactive B cells are deleted or functionally inactivated at the several different steps of maturation from immature B cells to antibody producing cells. These self-tolerance mechanisms appear to involve B cell apoptosis induced by signaling via the antigen receptor (surface immunoglobulin) or Fas. In mice prone to systemic autoimmune diseases such as bcl 2 transgenic, NZB or (NZB x NZW) F1 mice, antigen receptor-mediated B cell apoptosis is defective. In another autoimmunity-prone mice MRL/lpr, autoantibody production requires defects of Fas in B cells. These findings strongly suggest that the defects in B cell tolerance play an important role in the pathogenesis of systemic autoimmune diseases. PMID- 9200919 TI - [Superantigens and autoimmune diseases]. AB - Superantigens are potent immunomodulators derived from microorganisms such as bacteria, viruses and mycoplasmas. Their effects on immune systems are obtained through their binding both to outer portion of binding grooves of an MHC on antigen presenting cells and to non-recognizing structure of hypervariable region of T cell antigen receptors. X-ray crystallography revealed precise structures of some superantigens, a beta barrel domain as a ligand to MHC class II alpha chain and a beta grasp domain to TCRV beta chain. Superantigens induce not only T/B cell activation but also immunological tolerance through oligoclonal deletion and/or anergy. Contribution of superantigens to the pathogenesis of autoimmune diseases has been discussed since a superantigen, mycoplasma arthritis T cell mitogen revealed to be arthritogenic to mice, and the murine arthritis resembled human rheumatoid arthritis in the pathological findings. Rheumatoid arthritis as well as Kawasaki Disease, Sjogren syndrome, and multiple sclerosis is now well studied through the oligoclonal expression of TCR beta specificities on infiltrating T cells. Application of superantigens to the treatment of autoimmune diseases is also discussed. PMID- 9200918 TI - [Molecular mechanism of immunological recognition and the abnormality]. AB - Processing of non-self antigens is an initial step in the sequential immunoreactive system. However, the mechanism of the processing of endogenous antigens, which is presented with MHC (major histocompatibility complex)-class I molecules, has been remained without clarifying. Recently, proteasomes, functioning as a non-lysosomal, ATP/ubiquitin-dependent protease to degrade unnecessary proteins selectively, are thought to be a processing enzyme complex responsible for MHC class I-restricted antigen presentation. A major immunomodulatory cytokine, gamma-interferon (gamma-IFN), was found to regulate this processing system through two distinct mechanisms. First, gamma-IFN induced replacements of the proteasomal subunits X, Y and Z by LMP7, LMP2 and LMP10, respectively, producing "immunoproteasomes" that perhaps function more appropriate for the immunological processing of endogenous antigens. Second, the newly-identified proteasome activator, termed PA28, was induced greatly by gamma IFN. A relationship between the antigen presentation pathway and its abnormality is also discussed. PMID- 9200920 TI - [Molecular mimicry and mechanisms of autoantibody production]. AB - Molecular mimicry is defined as similar structures shared by products of dissimilar genes. This review article discusses a possible role of molecular mimicry in the production of autoantibodies. Antibodies reactive with products of bacterial and viral genes sometime cross-react with normal cellular proteins. Sera from patients with systemic autoimmune diseases show crossreactivity with some bacterial and/or viral gene products at a significant frequency. Identification of the structures of antigenic epitopes recognized by disease associated autoantibodies by expression cloning of autoantigen molecules and gene fragment expression revealed the amino acid residues that are shared by autoantibody-defined epitopes and microbial proteins. The presence of amino acid sequences shared between microbial proteins and autoantigens and the detection of antibodies in patients' sera that bind to the crossreactive epitopes suggest that immune responses to bacterial and viral infections may initiate the production of autoantibodies. Receptor-mediated phagocytosis of autoantigen molecules by crossreactive B cells and subsequent antigen presentation to helper T cells may facilitate the production of autoantibodies reactive with separate epitopes, if the autoantigen complex contains multiple B-cell epitopes and a shared T helper cell epitope. Analyses of the fine specificity of T helper cell epitopes on Friend murine leukemia virus env gene products revealed unexpected heterogeneity and redundancy of T cell responses even to a single epitope. This heterogeneity in T cell responses might play a role in the activation of self-reactive T helper cells through molecular mimicry. PMID- 9200921 TI - [Recent advances in animal models of autoimmune disease]. AB - Various autoimmune diseases can be induced in normal animals by simply manipulating the thymus/T cells. For example, elimination of CD25+CD4+ T cells, which constitute about 10% of peripheral CD4+ T cells of normal mice, leads to the development of various organ-specific autoimmune diseases, such as thyroiditis, gastritis, adrenalitis, insulitis, sialoadenitis, oophoritis, or orchitis. Reconstitution of CD25+CD4+ T cells prevents the autoimmune development. Elimination/reduction of CD25+CD4+ T cells induced by genetical manipulation of T-cell ontogeny or caused by environmental agents, such as virus infection, also causes similar autoimmune diseases in normal mice. These newly developed animal models of autoimmune disease will be useful for elucidating the mechanism of immunologic self-tolerance and pathogenetic mechanism of autoimmune disease. PMID- 9200922 TI - [Binding-peptide motifs of HLA class II molecules susceptible to autoimmune diseases]. AB - Recent advances in knowledge of crystal structures of MHC class II molecules has advanced understanding of the molecular basis for interactions between peptides and HLA class II molecules. Polymorphism of HLA class II molecules influences structures of peptides bound to HLA class II molecules. To elucidate mechanisms for statistical association between particular HLA class II alleles and susceptibility to autoimmune diseases, it is important to identify self peptides presented by disease-susceptible HLA class II molecules and triggering disease causative autoreactive T cells. In this study, we tried to identify self-peptides triggering autoimmune diseases including rheumatoid arthritis, insulin autoimmune syndrome, insulin dependent diabetes mellitus and infant-onset myasthenia gravis. Susceptibility to all of these diseases in the Japanese population are known to be strongly associated with particular HLA-DR-DQ haplotypes unique to Asians, and clinical features of some of these diseases are different between Caucasians and Asians including Japanese. We investigated differences in binding-peptide motifs between disease susceptible and non-susceptible HLA class II molecules and predicted candidates of autoimmune self-peptides carrying binding-motifs to disease-susceptible HLA class II molecules. Indeed the major epitope for insulin autoreactive CD4+ T cell was successfully identified by this strategy. We also found heterogeneity in immunogenetic background between Western type and Asian type of multiple sclerosis. Our data indicated that our strategy is useful to identify autoimmune self-peptides, and it is suggested that not only disease susceptible HLA class II but also self-peptides causing diseases are different between Caucasians and Asians. These differences may well correlate to different clinical manifestations of diseases between the two ethnic groups. PMID- 9200923 TI - [Peptide binding motif for HLA class I and autoimmune disease]. AB - Some HLA class I alleles are strongly associated with autoimmune diseases, such as HLA-B27 for ankylosing spondylitis (AS), and HLA-B51 for Behcet's disease. But it is not clear how the class I molecules play a role in induction of the diseases. Recent studies have shown that HLA class I associates with beta 2 microglobulin and one of a range of naturally processed short peptides. The peptides bind into a groove formed by two alpha helices and a beta-pleated sheet of HLA class I molecules. HLA class I molecules present peptides with allele specific motifs to T cells, and CTL recognize the peptides on HLA class I molecules. Further studies of peptides bound to HLA class I molecules are expected to identify peptides associated with autoimmune disease. PMID- 9200924 TI - [Detection of antigen specific T cell clonality in autoimmune diseases and epitope analysis of autoantigen-reactive T cells]. AB - In order to develop an ultimate therapy to overcome autoimmune diseases, it is very important to identify whether or not autoantigen-reactive T cells accumulate in such diseases. A novel method was established to analyze T cell clonality using a combination of reverse transcriptase-polymerase chain reaction of T cell receptor beta chain transcripts and single-strand conformation polymorphism (RT PCR/SSCP). Using RT-PCR/SSCP, clonal expansion of T cells were identified in NOD mouse, collagen induced arthritis model, rheumatoid arthritis, Crohn's disease, Sjogren's syndrome, bone marrow transplantation, pregnancy, malignant tumors, HTLV-I associated myopathy, and interstitial pneumonia. By using a tandem mass spectrometry or a lymphocytic proliferation assay, T cell epitopes were mapped in NOD mice, EAE, MCTD, and melanoma. PMID- 9200925 TI - [TCR repertoire of autoreactive T cells in autoimmune disorders]. AB - In human autoimmune diseases such as rheumatoid arthritis, Sjogren's syndrome, and multiple sclerosis, it has been clarified that autoreactive T cells play a crucial role in the generation of autoimmune disorders. Immunohistochemical studies have shown that most infiltrating lymphocytes are CD4 positive alpha beta T cells. Recent studies with polymerase chain reaction (PCR) provides evidence about the T cell receptor (TCR) V beta and V alpha genes on their T cells. Sequence analysis of the complementarity determining region 3 (CDR3), which is a central portion for recognition of antigens by T cells, indicates some conserved amino acid motifs, supporting the notion that infiltrating T cells recognize relatively few epitopes on autoantigen. PMID- 9200926 TI - [Costimulatory molecules in autoimmunity: role of CD28/CTLA4-CD80/CD86]. AB - Optimal T cell activation requires at least two signals. One is provided by the interaction between antigen-specific T cell receptor and antigen-MHC complexes. The other is provided by costimulatory signals. One best characterized costimulation is mediated by CD28/CTLA4-CD80/CD86 (CD28/B7). In the past decade, costimulatory signal mediated by CD28 has been shown to be critical to augment T cell proliferative response and effector function such as cytokine production. Recent intensive analysis of the CD28/B7 pathway have revealed unexpected means in which this pathway may be involved in the maintenance and breakdown of self tolerance. In vivo studies using antagonists or transgenes of this pathway reveals that inappropriate expression of CD80, ligand for CD28/CTLA4, could induce autoimmunity and blockade of CD28/B7 pathway could ameliolate several autoimmune disease models. Furthermore, CD80 and CD86 plays differential roles for the development of autoimmune disease since helper T cell development is differentially affected by the blockade of either. CD80 or CD86 in certain conditions. More recent studies demonstrated the crucial role of CTLA4 in negatively regulating T cell activation and autoreactivity. Taken together, this pathway play pivotal roles for autoimmunity, and manipulation of this pathway raises the possibility for controlling autoimmune diseases. PMID- 9200927 TI - [Role of TCR V alpha 24 J alpha Q+ T cells in autoimmune diseases]. AB - We investigated the role of T cell receptor (TCR) V alpha 24+ T cells in the pathogenesis of systemic sclerosis (SSc) and systemic lupus erythematosus (SLE). The invariant V alpha 24 J alpha Q was expanded and comprised 50-90% of the total V alpha 24 in healthy individuals. In contrast, patients with SSc and SLE showed the selective reduction of the invariant V alpha 24 J alpha Q with oligoclonal expansion of V alpha 24 TCR other than V alpha 24 J alpha Q. Because human V alpha 24 J alpha Q TCR is analogous to murine invariant V alpha 14 J alpha 281 TCR which is the major genotype of NK T cells, these results suggest that the selective reduction of T cells with invariant V alpha 24 J alpha Q TCR might play an important role of the generation of autoreactive T cells including T cells bearing other V alpha 24 TCR in autoimmune disease patients. PMID- 9200928 TI - [Cytokine gene expression in Th1 and Th2: its molecular mechanism and clinical implication]. AB - Activation of helper T-cells mediated by the T-cell receptor induces a series of biochemical events. Among them, both the activation of PKC/Ras- and CaM/CN mediated pathways play a central role in the signal transduction of cytokine gene induction. Cytokines produced by non-transformed Th1 and Th2 clones were classified into three groups, based on their signal requirement patterns for their expression. Closer examination using various stimulation conditions suggested that the balance between the activities of the two signaling pathways contributes to cytokine expression. Th1 and Th2 effector functions and their development are attributable to their coordinated and differential expression of cytokines. Clarification of the mechanisms of Th1/Th2 differentiation should lead to rational strategies for manipulating pathological immune responses. PMID- 9200929 TI - [An imbalance between Th1 and Th2-like cytokines in patients with autoimmune diseases--differential diagnosis between Th1 dominant autoimmune diseases and Th2 dominant autoimmune diseases]. AB - An imbalance between T helper cell (Th)1 and Th2-like cytokines has been described in several autoimmune diseases. Organ specific autoimmune diseases such as multiple sclerosis (MS) and inflammatory bowel diseases (IBD) are caused by Th1 dominant immune responses. On the contrary, systemic autoimmune diseases such as systemic lupus erythematosus (SLE) and Sjogren's syndrome(SS) are characterized by Th2 dominant imbalance of cytokine production. It might be useful for differential diagnosis among patients with various autoimmune diseases such as SLE, SS, IBD, and MS to measure the serum levels of cytokines such as IL 10, IFN gamma, and TNF alpha using ultrasensitive enzyme-linked immunosorbent assay system. PMID- 9200930 TI - [The mechanisms of oral tolerance]. AB - Oral or nasal administration of a single high dose or repeated mucosal delivery of low doses of proteins have been shown to induce systemic unresponsiveness in the presence of mucosal IgA responses. The induction of oral tolerance(or mucosally-induced tolerance) is mediated by T cells involved in the generation of active suppression, clonal anergy or clonal deletion. Studies of T helper(Th) cytokine responses have suggested that Th1- and Th2-type cells are involved in the induction of oral tolerance. Further, gamma delta T cells appear to be an important T cell subset for the regulation of oral tolerance. PMID- 9200931 TI - [Breakdown of B cell-tolerance and its genetic control]. AB - We examined Fas expression level and apoptosis-sensitivity of autoantibody producing B cells in systemic lupus erythematosus-prone (NZB x NZW)F1 mice. In young mice, B cells expressed little Fas. However, stimulation with anti-CD40 mAb up-regulated Fas expression, in which B cells were divided into two groups, one Fas(high), apoptosis-sensitive and the other Fas(low), apoptosis-resistant. In aged mice with overt SLE, a considerable proportion of B cells spontaneously expressed low level of Fas with apoptosis-resistant phenotype. Anti-DNA antibodies were virtually produced by apoptosis-resistant Fas(low) B cells in both young and aged mice. PMID- 9200932 TI - [V gene repertoire and pathogenic autoantibodies]. AB - The studies for autoantibody-associated V genes have failed to find any V gene which might be specific for pathogenic autoantibodies in humans; Namely, both normal subjects and patients have immunoglobulin V genes which can encode autoantibodies. Also single V gene can encode antibodies for both foreign antigens and autoantigens. However, some germline V genes such as V3-7 and VH4-21 are preferentially used for autoantibodies and may be regarded as prototype V genes for autoantibodies. Although somatic mutation in V genes is characteristic in IgG autoantibodies, 0-81 idiotype-positive IgM antibodies, which may be precursor B cells for pathogenic anti-DNA antibody also included many somatic mutations in VH genes, indicating an intrinsic abnormality in B cell development in SLE. Some studies also indicated an abnormal V gene repertoire in autoimmune states. These results may be attributed to dysregulation of autoantibody associated B cell development. PMID- 9200933 TI - [Analysis of B cell autoepitopes and mechanisms for autoantibody production in autoimmune diseases]. AB - One of the representative immunological disorders in systemic autoimmune diseases is production of autoantibodies. Recent studies were concentrated on B cell epitopes of intranuclear autoantigens using recombinant proteins. They revealed autoepitopes homologous to those on exogenous agents and multiple epitopes, which respectively indicated molecular mimicry and antigen-driven immune responses as a mechanism for autoantibody production. Further, some autoantigens are thought to be catalyzed in a disease-specific manner. These antigen-specific factors would contribute to the autoantibody production, cooperating with antigen-nonspecific factors such as polyclonal B cell activation, overexpression of cytokines and dysfunction of T cells and antigen presenting cells. PMID- 9200934 TI - [The mechanism of autoantibody production in systemic autoimmune diseases]. AB - The appearance of autoantibodies to nuclear autoantigens is the hallmark of systemic autoimmune diseases. The mechanism of the autoantibody production has remained elusive, though it is generally accepted that autoantigen-specific T cells drive the production. These autoantigen-specific T cells as well as autoreactive B cells can be found in peripheral blood from healthy people, suggesting that these autoreactive cells are regulated peripherally. In this review dealing with autoantibody production mechanism, the brief introduction of our approach employing mouse genetics is also included. PMID- 9200935 TI - [Endogenous retroviruses in autoimmune diseases]. AB - Endogenous retroviruses (ERVs) and those related genes are thought to be originated from integration of infectious retroviruses to germ cells or evolved from transposable genetic elements. Some of them can make biological effects on activities of hosts, by promoting the expression of flanking genes or producing certain regulatory proteins. If those mechanisms occur on cells in immune networks, immunological dysregulation including autoimmunity will develop. ERVs can also produce proteins serving as auto-antigens, followed by autoimmune responses. In this article, studies up to date on ERVs of animals and humans are shortly reviewed, related to immunological disorders to autoimmune diseases, showing some examples. PMID- 9200936 TI - [Pathomechanism of HTLV-I associated arthropathy and the role of tax gene]. AB - HTLV-I is known to be a causative agent for adult T cell leukemia. Recent studies revealed this virus is also related to several autoimmune disorders, such as arthropathy, myelopathy and Sjogren's syndrome. We studied etiology of HTLV-I associated arthropathy (HAAP), and found that tax is a causative gene for synovial proliferation and induction of immunogenicity. HTLV-I transgenic mice supported the etiopathological role of tax gene. Our results suggested that HAAP is considered to be a prototype of rheumatoid arthritis, and tax is a best tool for recognizing pathomechanism of rheumatoid arthritis. PMID- 9200937 TI - [Autoimmune rheumatic diseases associated with HIV infection and its pathogenesis]. AB - In autoimmune rheumatic diseases, retroviruses have been repeatedly discussed as important etiologic factors. However, despite a considerable amount of indirect evidence that retroviruses might indeed be involved in triggering or initiating autoimmune rheumatic diseases, clear cut direct evidence is still missing. Studies on autoimmune or rheumatic disorders associated with HTLV-I or HIV-I infection as well as new data from the autoimmune rheumatic mouse (MLR/1pr mouse) model might help to answer the questions how and what mechanisms retroviral infection may lead to autoimmune rheumatic diseases. From data obtained in patients with HIV-I infection, apoptosis and molecular mimicry to autoantigens opens new approaches to the study of rheumatic disease pathogenesis. PMID- 9200938 TI - [Systemic lupus erythematosus (SLE) and retrovirus]. AB - Retrovirus have repeatedly been suggested as a possible trigger mechanism for animal and human autoimmune disease. Recent reports indicate that not only exogenous, but also endogenous, retrovirus could play an important role to induce autoimmunity. Here we described the relationship between retrovirus and systemic lupus erythematosus. PMID- 9200939 TI - [Viral myocarditis and autoimmunity]. AB - The pathogenesis of viral myocarditis has not been clarified yet especially in the subacute and chronic stages. Recent reports have shown that myocarditis may be caused by autoimmune reactions. Autoantigens and autoantibodies have recently been reported in myocarditis and cardiomyopathy, and most of them may be induced by the cross-reaction between virus and cardiac tissue, molecular mimicry, or by the destruction of immune tolerance. However it is not clear how autoimmune reactions affect viral myocarditis. We have reported that cytokines are increased in the blood of patients with myocarditis and cardiomyopathy. Many kinds of cytokines modulate viral myocarditis in different ways in murine models. Nitric oxide is also recognized as an important factor in viral myocarditis. We have reported that drugs for heart failure modulate cytokine production. Classifying drugs from this point of view may be helpful for developing new therapeutic strategy for patients with myocarditis and cardiomyopathy. PMID- 9200940 TI - [Potential role of Epstein-Barr virus in Sjogren's syndrome]. AB - In this review, I discuss recent progress in our understanding of the role of Epstein-Barr virus on pathological conditions in Sjogren's syndrome (SS). In order to clarify the association of EBV in SS pathogenesis and further analyze the precise transcriptional mechanism of EBV reactivation, we assayed transcription of ZEBRA which is the fast transcribed EBV-encoded immediate early gene product and was an indispensable role in EBV reactivation. The ZEBRA expression was observed in the ductal epithelial cells and infiltrating B cells, and SS peripheral blood lymphocytes (PBL). On the other hand, to identify the inducer of EBV reactivation, we have used in vitro EBV reactivation model cell line. Screening of cDNA by mRNA differential display subtraction method, one clone, named "AK-1", was highly expressed in SS PBL. Nucleotide sequence analysis revealed that AK-1 was complete match with p300/CBP-associated factor (P/CAF). Furthermore, as another approach, we have partially cloned cDNA with a novel sequence containing a region homologous to the basic leucin zipper domain of CREB/ATF family by degenerate PCR. Our results together with recent reports, suggest the possibility that CREB family protein-p300-P/CAF ternary complex on ZEBRA promoter might be involved in ZEBRA transcriptional activation. PMID- 9200941 TI - [T cell vaccination--induction of anti-idiotypic immune response against TCR and shift of Th1/Th2 balance]. AB - T cell vaccination, originally contrived and coinned by I.R. Cohen is the injection of autoimmune pathogenic T cell line/clone or T cell receptor peptides in an attempt to induce anti-idiotypic regulation to treat autoimmune disease. Establishment of many T cell lines/clones from various autoimmune animal model and successful injection of these cells as T cell vaccine have been reported, although the exact mechanism for vaccination effect has not been elucidated. However, recent reports suggest that not the clonal deletion or anergy but the shift of Th1/Th2 balance of disease-related T lymphocytes may be involved in the effect of vaccination. PMID- 9200942 TI - [TCR peptide vaccination for autoimmune diseases: involvement of T-T interaction between autoimmune T cells presenting TCR peptide and CD4-CD8- regulatory T cells]. AB - TCR peptide vaccination has been recently regarded as a future therapy for autoimmune diseases. In spite of the potential value, its underlying mechanisms have not been clearly established. M. F. Kozovska, T. Tabira and I have recently analyzed the immune regulation triggered by V beta 17a-50-68 peptide vaccine in SJL/J mice and desciribed a novel T-T interaction between encephalitogenic T cells presenting TCR peptide and CD4-CD8- regulatory T cells (Journal of Immunology 157:1781, 1996). In this brief review, I discuss how anti-idiotypic regulation can be triggered by the exogenous TCR peptide, and what we have learned from the animal experiments with regard to the natural regulation in human autoimmune diseases such as multiple sclerosis. PMID- 9200943 TI - [Immune intervention by peptides having a MHC class II binding motif--application of MHC blockers to autoimmune disease models]. AB - Intervention of T cell activation and the treatment of autoimmune disease models by MHC class II binding peptides was reviewed in this article. Analog peptides derived from antigenic peptides were shown to inhibit T cell activation in vitro as well as in vivo either by T cell antagonism, T cell tolerance induction, or by MHC blockade. The induction of immune suppression by MHC blocker peptides was discussed in detail. Successful application of MHC blocker peptides in the treatment of experimental allergy encephalomyelitis (EAE), non-obese diabetic mice and collagen-induced arthritis models indicated that in vivo blocking of MHC class II molecules represents a promising approach for the prevention and possibly treatment of human autoimmune diseases. An approach in identifying non peptidic MHC blockers was also described. PMID- 9200944 TI - [Manipulation of costimulatory pathways in autoimmune disease]. AB - The past year has seen significant advances in our understanding of the role of costimulatory pathways in antigen-specific T cell activation and maintenance of self-tolerance. It has been suggested that the absence of costimulators on normal tissue cells could serve to induce self-tolerance and that inappropriate expression of costimulators on antigen-presenting cells (APC) could activate self reactive T cells, resulting in autoimmunity. CD28 on T cells and CD80 and CD86 on APC are key molecules in the maintenance and breakdown of anergy. CTLA-4Ig fusion protein, that binds to both CD80 and CD86 with high affinity and thereby prevents interaction of CD80/CD86 with CD28/CTLA-4, prevents or ameliorates several autoimmune disease in experimental animal models, supporting an importance of this pathway in the development of autoimmune diseases. However, the studies using specific monoclonal antibodies against CD80 and CD86 have shown different outcomes in individual autoimmune models. This suggests that the actual regulatory mechanisms of this pathway in autoimmunity is much more complex, because of the existence of two receptors (CD28 and CTLA-4) and two ligands (CD80 and CD86) and the opposite function of CD28 and CTLA-4 in T cell activation. Further investigation on physiological function of this pathway in vivo may help for developing rational therapeutic approaches manipulating this pathway. PMID- 9200945 TI - [Application of oral tolerance to the treatment of autoimmune diseases--active suppression and bystander suppression]. AB - Oral tolerance is a phenomenon in which an orally ingested antigen induces systemic hyporesponsiveness to the same antigen. If the mechanism can be applied to autoantigens, it could be a promising mode of antigen-specific immunomodulatory treatment for patients with autoimmune diseases. Multiple ingestion of low doses of antigen induces active suppression. Under this condition, suppression of autoimmune attack to target tissues is mediated by anti inflammatory cytokines such as TGF-beta, which are released from regulatory T cells triggered antigen-specifically. This type of oral tolerance, termed "bystander suppression", has one advantage of needing to know only the major components of the target tissue instead of exact epitopes of autoimmunity. Utilizing this mechanism, clinical trials of oral tolerance therapy are going on among the patients with multiple sclerosis, rheumatoid arthritis, and uveitis. PMID- 9200948 TI - [Specific inhibition of anti-DNA antibody production by an anti-DH ribozyme]. AB - In order to develop a new therapy for systemic lupus erythematosus, we designed a ribozyme (catalytic RNA) to specifically cleave the mRNA of anti-DNA-responsible VH genes and tested for their ability to regulate the production of autoantibody by human B cell clone O-81 expressing nephritogenic idiotypes. Based on a preliminary antisense experiments for O-81 VH gene, we determined the regions in O-81 mRNA, which would be the target site for the cleavege. The O-81 ribozyme specifically inhibited the production of anti-DNA antibody by O-81 cones. For application to clinical state, nuclease-resistance ribozyme has been expected. These results indicate to introduce ribozymes as a new method of therapy in autoimmune diseases. PMID- 9200947 TI - [Application of chimeric and humanized antibodies to autoimmune diseases therapy]. AB - Monoclonal antibodies (MAbs) have been extensively developed for treating autoimmune diseases such as rheumatoid arthritis, multiple sclerosis and intestinal bowl diseases. Recombinant DNA technology made it possible to manufacture chimeric and humanized MAbs, resulting in lower antigenicity, longer half-life in serum and higher biological activities compared to the original murine MAbs. Here, chimeric and humanized MAbs under clinical investigations in the field of autoimmune diseases are reviewed. At this time, non-deleting type of anti-CD4 MAb and anti-TNF alpha MAb have an attractive attention because of their excellent efficacy in the clinical trials. Although deleting type of anti-CD4 MAb failed to show the efficacy in double-blind phase III trials reproducibly, it should be re-evaluated on the administration dosage and duration. In the near future, MAbs to adhesion molecules and co-stimulatory molecules will be studied in clinics. PMID- 9200949 TI - [HIV infection and long-term non progressor]. AB - AIDS(acquired immunodeficiency syndrome) is caused by HIV(human immunodeficiency virus) that belongs to the lentivirus group. Generally, the length of time between HIV infection and development of AIDS is considered to be 10 years on average, but a few percent of infected persons-so-called long-term non progressor, do not develop AIDS even more than 10 years after infection. In long term non progressors, it has been described that 1) there is a low viral load, 2) isolated HIV is relatively non virulent type, 3) antibody to HIV do not enhance virus propagation, 4) type 1 cytokines is more dominant than type 2, and 5) antiviral activity of CD8+ cell is strong. Thus, in long-term non progressors, the immune-response actively suppress HIV proliferation, so that viruses are controlled at low level. PMID- 9200946 TI - [Monoclonal antibodies for treating autoimmune diseases]. AB - Monoclonal antibodies (mAbs) directed to selected cell surface antigens, cytokines and their receptors, and adhesion molecules have been shown to inhibit autoimmune responses in preclinial animal models. Therapeutic trials using the chimeric or humanized mAbs have already been undergoing in patients with rheumatoid arthritis (RA). Effects of anti-CD4 mAb are controversial, but anti TNF-alpha mAb reveals encouraging results. Although it is too early to determine the efficacy, mAbs are new hopeful therapeutic strategies for autoimmune diseases. PMID- 9200950 TI - [The gene therapy for a patient with ADA deficiency; report of the first gene therapy trial in Japan]. AB - Since the first gene therapy clinical trial for an ADA deficient patient was performed in September 1990, 10 ADA deficient patients have been enrolled in gene therapy clinical trial. We have been performing the first gene therapy trial in Japan for a 5 year boy with ADA deficiency since August 1995. Activated T cells from the patient's peripheral mononuclear cells were transduced by a retrovirus vector, LASN, which contained cDNA of human ADA gene, and re-infused to him intravenously after 7-11 days. We have already performed 10 cycles of the therapy for the patient. Here, we report the successful results of the gene therapy with laboratory and clinical evaluation. Furthermore, we overview the results of gene therapy for ADA deficient patients which were recently reported from 4 other groups. PMID- 9200951 TI - [Pathogenesis of IgA nephropathy: role of outer membranes of Haemophilus parainfluenzae antigens]. AB - IgA nephropathy is characterized by IgA deposits, predominantly in the glomerular mesangium and mesangial proliferative glomerulonephritis. Concerning its pathogenesis, several investigators suggest that the deposited IgA is an antibody to viral, bacterial, or dietary antigens. Such reports strengthen the possibility of a relationship between mucosal immunity and the pathogenesis of IgA nephropathy. We previously observed that Haemophilus parainfluenzae (HP) is more commonly isolated from the pharynx of patients with IgA nephropathy than from those with other diseases. We have also identified the glomerular deposition of the outer membranes of HP antigens (OMHP) and an increased serum concentration of IgA antibodies against OMHP in patients with IgA nephropathy. These findings suggest that HP has a role in the etiology of IgA nephropathy. PMID- 9200952 TI - [Clinical evaluation and perspective of charged particle therapy]. AB - In cancer radiotherapy, protons and heavy-ions share the same beneficial property of superior dose localization. In addition, heavy-ions are high-LET radiations and have increased biological effectiveness when compared with photons or protons. The potential indications for them are those diseases which are usually hard to cure with conventional radiations. At present there are 17 operating facilities world-wide for proton therapy, which in Japan is being conducted at the National Institute of Radiological Sciences (NIRS) and Tsukuba University (PMRC). Currently, Massachusetts General Hospital (MGH) in USA and the National Cancer Institute East Hospital in Japan are building a hospital-based proton facility, both of them will have a fixed energy cyclotron (230MeV). New plans for commencing proton therapy are also proposed by many other institutions in the world. As with heavy-ion therapy, clinical trials have been carried out since 1994 using carbon-ions generated by HIMAC (Heavy-ion medical accelerator in Chiba) at NIRS. The HIMAC is the world's only heavy-ion accelerator complex dedicated to medical use in a hospital environment. The Hyogo prefecture has just decided to build a similar ion therapy facility. As of February 1997, a total of 230 patients were treated in carbon-ion Phase I/II studies at NIRS. So far, carbon-ions appear to be effective in such cases as locally advanced tumors and non-squamous tumors. PMID- 9200953 TI - [Adipocyte differentiation and nuclear receptor]. AB - The roles of nuclear receptors in differentiation and function of adipocytes were reviewed and discussed. Expression of peroxisome proliferator-activated receptor (PPAR) gamma have been reported to be strongly induced during adipocyte differentiation and maintained in matured adipocytes. Forced expression of PPAR gamma converted NIH3T3 fibroblasts to adipocytes, indicating PPAR gamma regulates essential genes to obtain the adipocyte phenotype. Newly developed antidiabetic thiazolidinediones known as high affinity ligands for PPAR gamma improved insulin resistance. This finding suggests that PPAR gamma contributes regulation of insulin action. Several genes regulated by troglitazone, one of the most potent thiazolidinediones, in matured 3T3-L1 adipocytes-were obtained by differential display PCR method. Orphan receptors ROR alpha/gamma and Rev-ErbA which bind to the same response element are also induced during adipocyte differentiation but their function is still to be investigated. PMID- 9200954 TI - Gastric epithelial dysplasia. Is histological grading bearing any clinical relevance? PMID- 9200955 TI - Grading of gastric epithelial dysplasia. An interobserver study and analysis of diagnostic criteria. AB - One of the fundamental problems in the pathology of gastric epithelium is differentiation of reactive or regenerative proliferations, which are not precancerous from precancerous proliferations (i.e. dysplasia) and cancer. Diagnostic and interpretational difficulties, a need for a close cooperation between pathologists and clinicians and an attempt to more precisely assess gastric epithelial dysplasia prompted us to evaluate the usefulness of the current morphological criteria in the diagnosis and grading of gastric epithelial proliferations. The present study indicates that there are no sufficient grounds for grading of dysplasia, although the current morphological criteria permit establishment of a correct diagnosis. Therefore, the currently used three-grade classification of dysplasia may be successfully replaced with an easier two-grade classification or resigned totally. PMID- 9200956 TI - Retinoids and secosteroids induce gene expression in human colorectal carcinoma derived cells but the macroscopic-scale effects remain unpredictable. AB - Expression of a number of retinoid-responsive genes (hRAR alpha, CRABP I, CRABP II, MK cytokine) and secosteroid-responsive genes (hVD3R, Calbindin) was studied in in vitro model of human colorectal carcinoma by Relative RT-PCR. MK cytokine mRNA has been identified in human colonocytes for the first time. Proliferation of SW480 cells was inhibited by 5 microM all-trans retinoic acid and 5 microM 1 alpha, 25-dihydroxycholecalciferol; however, SW620 cells were not inhibited by all-trans retinoic acid. Unexpectedly, SW620 cells were stimulated by nanomolar concentrations of 1 alpha, 25-dihydroxycholecalciferol. In the latter case, no induction of gene expression was seen. Gene expression was induced in both cell types, whether there was a responsive element in the promotor region or not, suggesting that signal transduction to cellular nucleus did occur. Also, the Scatchard analysis for hVD3R receptor protein confirmed that the amount of the protein was modified under the treatment with both hormones; however, non-linear relationship between the amount of the mRNA and the protein was observed. In general, the genes responded differently to the treatment than it had been predicted. While this variability could be ascribed to the genetic instability, we hypothesize that instability in the cellular network of genes, mRNAs, and proteins is responsible for the observed effects. Due to the complexity, a microscopic-scale phenomenon such as gene expression cannot determine a macroscopic-scale process such as proliferation. This study provides a molecular background for retinoid/secosteroid chemoprevention of colorectal carcinoma; however, these hormones should be applied early to control premalignant lesions rather than advanced carcinomas. PMID- 9200957 TI - Ultrastructural changes in the small bowel epithelium after total gastrectomy. AB - The purpose of the study was to analyze ultrastructural changes in the small bowel mucosa in patients after total gastrectomy. We studied mucosal specimens obtained from 25 patients during control gastroscopy. The specimens were routinely processed for examination in transmission electron microscopy. Early after the operation (up to 6 months) we observed marked inflammatory reaction, disordered architecture of the small bowel mucosa epithelium, the presence of dysplasia-like changes and foci of dysplasia. Later on the structure of the mucosa returned to normality. Only a few dysplastic changes were seen. No relationship was found between altered epithelial structure and type of operation. In conclusion, the epithelium of the small bowel does not transform to a gastric type epithelium. PMID- 9200958 TI - Ki-67 as a marker of proliferation activity in tumor progression of recurrent gliomas of supratentorial localization. Immunocytochemical quantitative studies. AB - Quantitative study of labeling index (expressed in per cent of positive cells to all tumor cells) of Ki-67 (Ki-67LI), a marker of tumor proliferation, was performed in a group of 37 primary glial tumors and their recurrences. Three cases were entirely negative (no positive cells were found). Mean Ki-67LI for all primary tumors was 19.2% and for their recurrences-20.1%. There was no significant difference between tumors with longer (exceeding 1 year) and shorter survival between first and second surgery. In 17 cases recurrent tumor was of the same grade and in 16 cases malignancy at second surgery was higher than in primary tumor. Among tumors whose malignancy increased between first and second surgery as well as among tumors with the same grade at first and second surgery there were cases with increased and decreased Ki-67LI comparing samples from primary and secondary (recidiving) tumors. It is however, noteworthy that there was no tumor with markedly (at least twofold) decreased Ki-67LI and increased malignancy. The present findings indicate that there is still no simple answer to the question of prognostic role of Ki-67 in gliomas. PMID- 9200959 TI - Examination of DNA-ploidy in melanocytic nevi cells using video-imaging cytometry. AB - DNA content in 61 different types of melanocytic nevi was determined by video imaging cytometry. The nevi were selected for this study because of melanoma risk associated with each nevus, which is 20-50% according to different authors. DNA ploidy was detected in paraffin embedded and fresh tissue material of each patient and the results were comparable. The sample preparation process and video imaging method were presented in this study, 47 (77.0%) lesions exhibited diploid cell populations, 13 (21.3%) aneuploid and 1 (1.7%) tetraploid cell population. A significant correlation was observed between DNA ploidy measured in fresh tissue and retrospective material. The results indicate the presence of abnormal DNA content in some of the lesions. PMID- 9200960 TI - Mesangiocapillary glomerulonephritis type III and idiopathic membranous glomerulopathy. Morphometric comparison of the deposits and glomerular basement membrane including clinico-morphological correlations. AB - Fifteen renal biopsy specimens from patients with idiopathic mesangiocapillary glomerulonephritis type III (MCGN-III) and fifteen from patients with primary membranous glomerulopathy (MGN) for whom both light and electron microscopy as well as immunofluorescence microscopy and full clinical data were available were examined quantitatively and compared with six normal controls. Morphometric investigations of electron micrographs were performed by means of a computer image analysis system to compare glomerular basement membrane (GBM) thickness and the electron-microscopic density of the deposits in MCGN-III and MGN as well as to study whether these parameters could correlate with the clinical data. The study revealed that GBM thickness and the electron-microscopic density of the deposits were significantly-increased in MGN in comparison with MCGN-III. It should be also noted that the degree of proteinuria was closely correlated with the density of the deposits in both MCGN-III and MGN groups. While the present morphometric analysis of glomerular ultrastructure has not yet elucidated the nature of hematuria in MCGN-III and MGN, our results suggest the location rather than the density of the deposits may play a role in the process. PMID- 9200961 TI - Cholesterol crystals, smooth muscle cells and new data on the genesis of atherosclerosis. AB - The histologic appearance of atherosclerosis has been well described but its pathogenesis is still vigorously debated. The purpose of the present study is to clarify the stimuli for smooth muscle cells (SMC) migration and proliferation as well as the way of cholesterol crystals (CC) generation. We performed postmortem ultrastructural analysis of myocardial samples obtained from 45 patients (33 males, 12 females, age range 18-85) who died from different diseases, mainly from acute myocardial infarction-MI (37 cases). Tissue was taken by transthorax express-necropsy method immediately after patients' death in the clinic. In myocardial infarction cases intact zones of the heart were examined. We have found a few foci of modified SMC proliferation in the periarteriolar space around necrotic cellular debris in elder patients. Lymphocytes, myofibroblasts and some leukocytes infiltrated those areas. The modified SMC phagocyted necrotic material and formed secondary lysosomes, inside which from one to three CC originated. In parallel with the reduction of necrotic mass inside the lysosomes, CC size increased. In the final phase the CC were discharged from the SMC into the interstitium. After exocytosis a tendency for the CC to accumulate was observed. They formed clusters consisting of 20-30 crystals. Most of the CC were of typical needle-like or rhomboid shape. Modified SMC were producing not only CC but also collagen and elastin. This study indicates that atherosclerotic process in the myocardium is connected with the appearance of modified SMC inside which CC are generated. PMID- 9200962 TI - Synchronous carcinoma and primary MALT-lymphoma of the stomach. A report of two cases. AB - A synchronous presentation of an adenocarcinoma and a primary low grade B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) in the stomach is reported in a 73-year-old woman and a 55-year-old man. The diagnosis was based on microscopic examination of surgical specimens with immunohistochemistry. A possible etiology of the simultaneous presence of these two neoplasms in the stomach is discussed on the basis of our own material and review of the literature. PMID- 9200963 TI - Pulmonary infiltrates with masto- and lymphocytosis in BALF associated with arechine. AB - Pulmonary infiltrates developed in a 40-year-old man while receiving arechine. The hyperplasia of mast cells, hyperlymphocytosis, a predominance of cytotoxic T cells of the CD8 type with the very low CD4/CD8 ratio in BAL were observed. The chest roentgenogram findings resolved within a month after discontinuing arechine therapy. A hypersensitivity reaction with the onset of lung fibrosis due to arechine seemed to be the likely cause. PMID- 9200964 TI - Simvastatin-induced myopathy in a patient treated for hypercholesterolemia. Morphological aspects. AB - Functional and histological changes in skeletal muscle developing during hypolipemic therapy, especially with 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase inhibitors are rare. This paper reports a case of simvastatin induced myopathy confirmed histopathologically and ultrastructurally. PMID- 9200965 TI - Systolic function impairment in higher erythropoietin requirement patients in hemodialysis. AB - Circulatory adaptation to hemodialysis (HD) depends on several inter-related factors, due to the particular patient and to technical procedures. The cardiac effects of nutritional status and the improvement that erythropoietin (r-HuEPO) treatment, through the reduction of anemia, can determine, were reported; r-HuEPO requirements are quite different, in different patients, while achieving similar haemoglobin end-point. Thirty-four patients on r-HuEPO and 11 patients not treated, all in hemodialysis, were studied by echocardiography at beginning and at the end of 3 consecutive dialysis sessions. Ejection fraction increased with hemodialysis in all patients of the control group. A slight and not significant decrease of ejection fraction was observed, as average, in the r-HuEPO treated patients. But, more in detail, 20 patients on r-HuEPO showed a worsening of systolic function with hemodialysis; in the other 14 patients on r-HuEPO, as in not treated control patients, systolic function improved. A significant difference between the two sub-groups on r-HuEPO was the higher drug dose requirement, associated with a slightly lower haemoglobin concentration, both observed in patients with decreased ejection fraction after hemodialysis. Moreover, a close relationship between higher r-HuEPO requirements and worsening of systolic function was observed. This trend is not associated with adequacy of dialysis and malnutrition. Patients with higher r-HuEPO requirement share a worse cardiac adaptation to hemodialysis and slightly lower haemoglobin levels. PMID- 9200966 TI - [An unusual location of sporotrichosis in a subject with a carcinoma]. PMID- 9200968 TI - Progress towards leprosy elimination. PMID- 9200967 TI - Role of lipopolysaccharide and related cytokines in Helicobacter pylori infection. AB - Helicobacter pylori is a gram-negative bacterium which accounts for the development of chronic gastritis and peptic ulcer in man. In this review, emphasis has been laid on the role of lipopolysaccharide (LPS) of the H. pylori cellular wall in the pathogenesis of gastroduodenal disease. H. pylori LPS exhibits a reduced endotoxic potency in terms of pyrogenicity, lethality, toxicity, mitogenicity, cytokine (CK) release and chromogenic limulus amebocyte lysate assay. This low biological activity of the LPS could be ascribed to the underacylation and underphosphorylation pattern of the lipid A backbone. However, also LPS core structures seem to contribute to the biological activity of the molecule. Despite this low immunological potential, an array of proinflammatory CKs are produced both in vitro and in vivo following stimulation of mucosal cells with H. pylori organisms. It is likely that LPS plays a major role in triggering interleukin (IL)-8, IL-1 and tumor necrosis factor-alpha production from both epithelial cells and macrophages. Finally, the lower immune response elicited by H. pylori LPS in comparison with other enterobacterial LPS may represent an escape mechanism from the host immunosurveillance exerted by this bacterium, thus allowing its survival and persistence in the gastric niche. PMID- 9200970 TI - Multifactorial self-concept and delinquency in Australian adolescents. AB - Self-concept is an integral part of identity development for adolescents. The present study was designed to test Australian adolescents self-evaluations regarding delinquent behaviors. The measures used (Self-Report Delinquency Scale [Mak, 1993] and Personal Descriptive Scale) successfully differentiated nondelinquents, noninstitutionalized delinquents, and institutionalized delinquents. The relative self-concept scores of the groups indicate that the more serious the delinquent behavior, the more negative the self-concept. PMID- 9200969 TI - An unusual manifestation of Meckel's diverticulum: bleeding and perforation--a case report. AB - Meckel's diverticulum is the most common congenital abnormality of the gastrointestinal tract. Bleeding of the diverticulum from a peptic ulcer due to ectopic gastric mucosa makes up 25% of all complications in the disease. We report a case of a 20 year old male with chronic anemia, severe hematochezia and perforation due to Meckel's diverticulum. Segmental small bowel resection was performed. Difficulty of preoperative diagnosis is discussed, indication for incidental diverticulectomy is established and the literature reviewed. PMID- 9200971 TI - Psychological reaction to perceived erasure of community boundaries. AB - The reaction of residents of a small district in the Northeast of England to the perceived compromising of existing community boundaries was examined in the light of Tajfel and Turner's (1986) social identity theory. Social identity theory was found to provide a useful framework for explaining specific psychological reactions of residents and spokespersons to the perception of ingroup status inferiority. PMID- 9200972 TI - Cooperativeness and bully/victim problems among Australian schoolchildren. AB - The relationship was examined between the self-reported cooperativeness of Australian secondary-school students and their involvement in peer abuse at school, both as bullies and as victims. An 18-item Likert-type measure, the Cooperativeness Scale, was developed, and its reliability and concurrent validity were supported by the results of its application to two samples of Australian students (N = 176 and N = 763, respectively) attending different coeducational secondary schools, the first in a predominantly middle-class area and the second in a lower class socioeconomic area. At both schools, girls scored higher in cooperativeness than boys. Students at the second school also anonymously completed multiple measures of the extent of their involvement during the current year in bullying, victimization, or both. As predicted, correlations and multiple regression analyses supported the hypothesis that relatively low levels of cooperativeness were characteristic, not only of both boys and girls who engaged in bullying, but also, to a lesser extent, of those who were frequently victimized by their peers at school. PMID- 9200973 TI - Gender differences in extraversion, neuroticism, and psychoticism in 37 nations. AB - Mean gender differences on Eysenck's three personality traits of extraversion, neuroticism, and psychoticism were collated for 37 nations. Women obtained higher means than men on neuroticism in all countries, and men obtained higher means than women on psychoticism in 34 countries and on extraversion in 30 countries. The relation between the magnitude of the gender differences and per capita incomes was not significant for any of the three traits. PMID- 9200974 TI - Age and gender differences in the self-esteem of Chinese children. AB - A Chinese version of the Self-Description Questionnaire 1 (SDQ-1; Marsh, 1988) was used to investigate age and gender differences in a sample of 303 male and 296 female 10-year-old children and 116 male and 116 female 13-year-old children attending typical Beijing public schools. Significant Age x Gender interaction effects were found on all 8 SDQ-1 scales. Main effects for age were found on the Physical Abilities, Reading, and School subscales and for gender on the same three subscales plus Peer Relations. Further analysis indicated that the older girls tended to report significantly lower self-esteem than both the younger girls and older boys in the areas of physical abilities, reading, mathematics, and general self-concept. The boys reported more positive self-perceptions on most nonacademic self-scales, but both the older boys and older girls reported less favorable self-esteem than their younger peers on the scales for reading and school in general. PMID- 9200975 TI - Person perception as a function of marital status and age. PMID- 9200976 TI - Distribution and function of gonadotropin-releasing hormone (GnRH) in the teleost brain. PMID- 9200977 TI - Identification of a vanadium-associated protein from the vanadium-rich ascidian, Ascidia sydneiensis samea. AB - Ascidians are known to accumulate vanadium in their blood cells (vanadocytes) at extremely high levels which correspond to about 10(6) to 10(7) times the levels of vanadium ions in seawater. The route for the accumulation of vanadium ions from the outside environment into the blood system in ascidians has not yet been discovered. In the present experiments, using a combined technique of anion exchange column and atomic absorption spectrometry, we first extracted a vanadium associated protein (VAP) from the blood cells of the ascidian Ascidia sydneiensis samea. VAP was estimated to associate with vanadium at an approximate ratio of 1 mol:16 mole. SDS-PAGE and a polyclonal antibody against VAP (anti-VAP) revealed that VAP is composed of at least two types of peptides estimated to be 12.5 kDa and 15 kDa with a minor peptide of 18 kDa and that VAP is localized in the cytoplasm of the vanadocytes. PMID- 9200978 TI - Finding of the same antigens in the polychaete, Pseudopotamilla occelata, as those in the vanadium-rich ascidian, Ascidia sydneiensis samea. AB - The polychaete Pseudopotamilla occelata is the first animal revealed to contain high levels of vanadium besides ascidians. The present experiment disclosed that P. occelata has the same antigens with those in the ascidian Ascidia syndneiensis samea, which were recognized by two types of antibodies, a polyclonal antibody against vanadium-associated proteins extracted from blood cells and a monoclonal antibody against vanadocytes in the vanadium-rich ascidian A. sydneiensis samea. There is, therefore, a possibility that similar mechanism works on the accumulation of vanadium between the Polychaeta and the Ascididae. PMID- 9200979 TI - Delayed response of QM- and DA/DAPI-fluorescence in C-heterochromatin of the small Japanese field mouse, Apodemus argenteus. AB - The small Japanese field mouse Apodemus argenteus has the diploid chromosome number of 46, carrying rather large centromeric C-heterochromatin in most of the 44 autosomes and a large amount of C-heterochromatin in the sex chromosomes: the largest subtelocentric X was heterochromatic in almost two-fifth (whole short arm and proximal part of the long arm) of its entire length and the medium-sized acrocentric Y was totally heterochromatic. The C-heterochromatin (C-positive) areas, other than those of the Y and smallest three pairs, had a unique property of "delayed QM-fluorescence", which has not been reported to-date, showing dull QM-fluorescence immediately after exposure to blue light (BL), but gradually turning to bright fluorescence in a few minutes. The fluorescence intensity gradually decreased after attaining its peak, and finally became extinct. A similar pattern of fluorescence was also obtained in DA/DAPI-stained-X chromosome C-heterochromatin, but not in autosomal C-heterochromatin. No such dull-to-bright transition of QM-fluorescence could be obtained by CMA staining, for which the C positive areas were apparently negative even after overexposure to BL. These facts indicate that the C-positive areas of A. argenteus showing dull-to-bright transition of QM-fluorescence contain A-T rich DNA. The delayed QM-fluorescence was found only in A. argenteus, in thirteen mammalian species so-far examined. Furthermore, this unique property of QM-fluorescence could be artificially altered to non-delayed ordinary type of fluorescence by sequentially pretreating the fixed chromosomes with hydrochloride and barium hydroxide solutions. The cytological implication of the delayed fluorescence in the C-heterochromatin of A. argenteus is briefly discussed. PMID- 9200980 TI - Isolation of a cDNA encoding a chitinase family protein from cuticular tissues of the Kuruma prawn Penaeus japonicus. AB - To identify and characterize a chitinase related to molting in the Kurumia prawn Penasus japonicus, we searched for chitinase-encoding cDNAs expressed in cuticular tissues. Using two degenerate oligonucleotide primers derived from the two conserved regions of the chitinase protein family, a RT (reverse transcription)-PCR product was obtained. This product was used as a probe to screen a cDNA library from a mixture of the tall fan and blade-two tissues which consist mainly of chitinous exoskeleton and underlying epidermis. A positive cDNA clone was analyzed for the sequence. This clone contains an open reading frame for a protein (named Pjchi-2) of 527 amino acids which exhibits sequence similarity to known chitinases. A typical signal sequence could not be found in the Pjchi-2 sequence. Significant accumulation of Pjchi-2 mRNA was detected in the mixture of the tall fan and blade prior to molting, whereas the transcript level was much lower during the intermolt stage. This observation suggests that Pjchi-2 plays a role in molting. The mRNA was not detected in the hepatopancreas. This expression pattern of Pjchi-2 makes a contrast to that of Pjchi-1 which encodes another chitinase family protein in P. japonicus, and is expressed in the hepatopancreas but not in the tall fan or blade. PMID- 9200981 TI - Cloning and sequencing of gene coding for a periplasmic 5.4 kDa peptide of the macronucleus-specific symbiont Holospora obtusa of the ciliate Paramecium caudatum. AB - We purified a 5.4 kDa peptide which is present in the intermediate and infectious form, but not in the reproductive form of a symbiotic bacterium Holospora obtusa of the ciliate Paramecium caudatum. Sequencing of its gene revealed that it encodes a peptide composed of 49 amino acids, and that the peptide is preceded by a putative signal peptide of 19 amino acids. We determined the transcription start point for the gene by primer extension analysis, indicating that the transcription starts with a G nucleotide located 33 nucleotides upstream from the translational initiation codon. Northern blot hybridization showed that the gene is highly expressed in the intermediate form, a transitional stage from the reproductive to infectious form of the bacterium. Immunoelectron microscopy with anti-5.4 kDa peptide antiserum revealed that the 5.4 kDa peptide is localized in the periplasm of the infectious form. PMID- 9200982 TI - Primary structure of mouse actin-related protein 1 (Arp1) and its tissue expression. AB - Different types of actin-related proteins which constitute an actin-superfamily together with conventional actin have recently been described (Mullins et al., 1996). Among them, Arp1 exhibits the highest homology with conventional actin. With the aim of clarifying the cellular function of Arp1 in mammalian cells, we cloned the cDNA encoding mouse alpha-Arp1, one of the variants of Arp1, from a mouse diaphragm cDNA library; two types of alpha-Arp1 cDNAs, which are probably generated by alternative RNA splicing from a single gene, were obtained and the entire sequences were determined. They differed only in the presence or absence of an insertion of 1.3 kb in the 3'-non-cooling region but shared a common open reading frame. The deduced amino acid sequence was identical with that of human alpha-Arp1. Northern blot analysis showed that the alpha-Arp1 mRNA corresponding to the longer cDNA is transcribed not only in various non-muscle tissues but also in muscle tissues, while the transcript corresponding to the shorter one becomes expressed only in skeletal muscle as development progresses. It is suggested that alpha-Arp1 may play some role in muscle, as judged by the significant level of its expression. PMID- 9200983 TI - Cellulose digestion in the wood-eating higher termite, Nasutitermes takasagoensis (Shiraki): distribution of cellulases and properties of endo-beta-1,4-glucanase. AB - beta-Glucosidase [EC 3.2.1.21] and endo-beta-1,4-glucanase [EC 3.2.1.4] activities were measured in the wood-eating higher termite Nasutitermes takasagoensis. beta-Glucosidase activity was present mainly in the salivary glands (66.7%) and midgut (22.2%), whereas endo-beta-1,4-glucanase activity was detected mainly in the midgut (90.1%). Specific activity of endo-beta-1,4 glucanase was also the highest in the midgut, indicating that cellulose is digested in the midgut. The major endo-beta-1,4-glucanase component of N. takasagoensis was purified from whole termites by gel filtration on Sephaoryl S 200 HR, Superdex-75 and hydroxyapatite column chromatography. Subsequently, the endo-beta-1,4-glucanase activity from a crude midgut extract was eluted in an identical volume (Kd = 0.68) to that from whole termites, suggesting the purified endo-beta-1,4-glucanase is identical to that in the midgut. The molecular weight of the purified endo-beta-1,4-glucanase was 47 kDa, and its specific activity was 1,200 units/mg. The optimal pH and temperature were 5.8 and 65 degrees C, respectively. The Km and Vmax values on carboxymethyl cellulose were 8.7 mg/ml and 2,222 units/mg, respectively. The purified endo-beta-1,4-glucanase hydrolyzed cellopentaose to cellotriose and cellobiose, and cellotetraose to cellobiose and a trace of cellotriose and glucose, but cellotriose and cellobiose were not hydrolyzed. The activity and stability on pH and temperature of the purified endo beta-glucanase are prominent among those from various organisms. PMID- 9200986 TI - Possible involvement of folliculo-stellate cells in the differentiation of muscle fibers during monolayer culture of pituitary cells. AB - A major objective of the present study was to examine the possibility that non granular folliculo-stellate (FS) cells in the rat anterior pituitary are involved in the myogenesis that occurs during pituitary cell culture. Enzymatically dissociated anterior pituitary cells were fractionated by use of the Percoll gradient method. The proportion of FS cells was 5.8% on average before cell fractionation. After employing the Percoll gradient procedure, FS cells were enriched to a ratio of 12.2%. Three of five cell fractions were separately cultured, and the incidence of striated muscle fibers was quantitatively investigated. There was a good correlation between the numbers of muscle fibers and the proportions of FS cells in the fractions obtained from the Percoll gradient. These results suggest that FS cells are the cells that transform into striated muscles in pituitary monolayer cultures. PMID- 9200984 TI - Cell-cell contact down-regulates expression of membrane type metalloproteinase-1 (MT1-MMP) in a mouse mammary gland epithelial cell line. AB - Membrane type matrix metalloproteinase (MT-MMP), which possesses a C-terminal transmembrane domain, is expressed on the cell membrane (Sato et al., 1994, Nature 370: 61-65). It was suspected, therefore, that the expression of MT-MMPs might be regulated by cell-cell interactions. We examined the patterns of MT1-MMP expression in a mouse mammary gland epithelial cell line, HC11, which is capable of responding to prolactin in vitro. HC11 cells form well-differentiated monolayer of cuboidal epithelium at confluence. During the log growth phase, cells which are well dispersed and seemingly migrating actively, or located at the periphery of small colonies, reacted strongly with an anti-MT1-MMP antibody, whereas no MT1-MMP immunoreactivity was detected in the cells which established cell-cell contact with adjacent cells. At confluence, the HC11 cells lost MT1-MMP immunoreactivity completely. Northern blot analysis revealed that MT1-MMP mRNA is present at a high level in HC11 cells during the log phase of growth. Although MT1-MMP immunoreactivity disappeared by the 1st day confluence was reached, the decline of MT1-MMP mRNA levels started only a few days later. The discrepancy in the timing of decrease of MT1-MMP protein and that of the transcripts suggests the presence of translational control mechanisms for MT1-MMP expression during cell-cell interaction. PMID- 9200985 TI - Stability of chicken troponin T expression in cultured muscle cells. AB - Cells prepared from chicken skeletal muscles of early developmental stages were cultured to study their troponin T isoform expression, using antisera specific to fast- and slow-muscle-type isoforms, and compared with the cells from later stages described in the previous study (Mashima at al., 1996). We found that cultured myogenic cells from chickens and chick embryos could be classified, as in the previous study, into two types, fast type and fast/slow type in which fast and slow-muscle-type isoforms were coexpressed. Ratios of these two types of muscle cells varied depending on their origins and developmental stages, and fast/slow type cells were in the majority at early stages. Since two distinct populations of cells committed to myogenic cell lineages were supposed to give rise to the two types of myotubes, we investigated the intrinsic stability of troponin T expression of the cultured myogenic cells using the serial subcloning method. The results of clonal analysis suggested that the expression pattern of troponin T isoform in cultured muscle cells is stable and that myogenic cell lineages play an important role in giving rise to different muscle types. PMID- 9200987 TI - Enhancement of the receptor binding and Nb2 proliferation activities of rat prolactin by site-directed mutagenesis. AB - To investigate the roles of the amino acids of rat prolactin (rPRL), the structure of which is presumed to consist of an antiparallel, four-alpha-helix bundle, mutations constructed by site-directed mutagenesis were assayed in terms of their receptor binding and Nb2 cell proliferation activities. Replacement of P64L (replacing proline at position 64 with leucine) and K67E, which are located in the long loop region between helices 1 and 2, produced drastic decreases in the binding and proliferation activities. Mutations at D91 and E118 in the second and third helices, respectively, resulted in increased Nb2 cell proliferation activity with a slight increase in receptor binding activity. Mutations at L81 in the second helix and Y145 and W148 in the second loop between helices 3 and 4 produced no marked changes. Mutation at D158N in helix 4 markedly increased receptor binding activity with a slight loss of Nb2 cell proliferation activity, although two other mutation, D158H and D158R, produced a decrease of receptor binding activity without notable changes in Nb2 cell proliferation activity. These results demonstrate that P64 and K67 are crucial for PRL function while replacement of D91, E118 and D158 possibly leads to functional enhancement. PMID- 9200988 TI - Effects of estrogen and dopamine agonists on the expression of argyrophilic nucleolar organizer regions in prolactin cells of rats. AB - The number of nucleolar organizer regions reflects nuclear and cellular activity, such as the proliferation and differentiation of cells. Prolonged administration of estrogen (E2) induces the hyperplasia of prolactin (PRL) cells and the development of PRL-secreting pituitary tumors in rats. Dopamine agonists are known to reduce the effects of E2 treatment. The purpose of this study was to investigate whether the changes of argyrophilic nucleolar organizer regions (AgNORs) in PRL cells are related to circulating levels of PRL or to the proliferative activity of PRL cells during the administration of stimulatory (E2) or inhibitory (dopamine agonist) treatment in rats. E2 increased the size and number of AgNORs per nuclear profile in PRL cells in rats. Bromocriptine and cabergoline, which are dopamine agonists, each reduced the number and size of AgNORs in PRL cells treated with E2 for 10 weeks. In rats treated with E2 alone or dopamine agonists followed by E2, the number and size of AgNORs were correlated more closely with serum levels of PRL than with the proliferative activity of PRL cells. However, neither the number nor size of AgNORs in PRL cells was related with these parameters in different ages of the control. The number and size of AgNORs may be useful in evaluating the secretory activity of pituitary cells during the administration of stimulatory or inhibitory agents. PMID- 9200989 TI - Effect of hormones on expression of prolactin receptor messenger ribonucleic acids in pancreatic islets of adult female mice in vitro. AB - We studied the effects of hormones on expression of prolactin receptor (PRL-R) mRNA in pancreatic islets of adult female mice in vitro. We quantified mRNA expression in small amounts of the islet tissue by competitive PCR and one-sided competitive PCR. Fifty pancreatic islets from adult female mice were cultured in a wall for 4 days with or without ovine prolactin (PRL), bovine growth hormone or estradiol-17 beta. PRL (1 microgram/ml) significantly increased the insulin secretion and the amount of PRL-R mRNA relative to that of beta-actin mRNA. Growth hormone (1 microgram/ml) also increased the relative amount of PRL-R mRNA, although it did not significantly increase the insulin secretion. Neither insulin secretion nor the relative amount of PRL-R mRNA was affected by estradiol (100 ng/ml). The ratio of the short form to the long form of PRL-R mRNA was not altered by these hormones. The present observation that PRL increased PRL-R mRNA expression in pancreatic islets thus suggests the possibility that PRL up regulates the tissue sensitivity to PRL itself during lactation. PMID- 9200990 TI - Phylogenetic position and geographic differentiation of the Japanese dormouse, Glirulus japonicus, revealed by variations among rDNA, mtDNA and the Sry gene. AB - The Japanese dormouse, Glirulus Japonicus, is the only extant lineage that represents this genus and it has been classified as a single species distributed on the three main islands of Japan, namely Honshu, Shikoku and Kyushu. However, individuals collected from Fukui, Wakayama and Kochi Prefectures (southwestern part of Japan) yielded distinctly different profiles of restriction fragments of the nuclear ribosomal DNA (rDNA) spacer from those collected from Yamanashi and Nagano Prefectures (central Japan). The estimated sequence divergence between the two groups was 2.8% on average, which corresponds to a putative divergence some two million years ago. Representing mitochondrial DNA (mtDNA) sequences, 402 bases of cytochrome b gene were determined by direct sequencing and the estimated extent of the sequence divergence between the two groups was 6.5-7%. Differences between the two geographic groups were also substantial in the sequences of about 300 base-fragments from the Y-linked, sex-determining locus, Sry. To assess the phylogenetic relationships between the Japanese dormouse and members of the family Myoxidae, we compared sequences of mitochondrial 12S rRNA gene of Japanese dormice with those of the forest dormouse (Dryomys nitedula) and the common dormouse (Muscardinus avellanarins), two continental genera thought to be closely related to the genus Glirulus. The results showed that the sequences from Japanese dormice were distinct from any sequences of the two continental species and the extent of the differences were somewhat similar to that between the rat (Rattus norvegicus) and the hamster (mesocricetus auratus). PMID- 9200991 TI - S-HAM induction chemotherapy with or without GM-CSF in patients with high-risk myelodysplastic syndromes. AB - Thirty-one adult patients with high-risk myelodysplastic syndromes (MDS) were enrolled in a prospective randomized double-blind placebo-controlled trial evaluating the efficacy of sequential high-dose Ara C/mitoxantrone chemotherapy with or without GM-CSF. GM-CSF or placebo was given subcutaneously once daily at a dose of 250 micrograms/m2 starting 48 h prior to chemotherapy and continued until neutrophil recovery. This design allowed us to investigate the role of GM CSF as a priming factor for the leukemic clone, as well as its effect on the recovery of normal hematopoiesis. Twenty-eight patients are currently evaluable for response. The patients reached a complete remission (36%), eight patients had persistent MDS (29%), and ten patients died within 6 weeks after the onset of treatment (early death). Infectious complications during cytopenia were the major cause of death (8/10). Median time to complete hematologic recovery (neutrophils > 500/microliter and platelets 20,000/microliter) and time to neutrophil recovery above 1500/microliter was 29 and 35 days, respectively. Median remission duration was 190 days (6.4 months). Analysis of prognostic subgroups showed a low CR rate (25%) and a high early-death rate (44%) in patients > 55 years of age, suggesting that the intensified treatment approach should be limited to younger patients. No data concerning the influence of GM-CSF on response to chemotherapy or duration of neutropenia are presently available. PMID- 9200992 TI - Association between trisomy 8 and the immunophenotype of blast cells from acute leukemias secondary to a myelodysplastic syndrome or chronic myeloproliferative disorders. AB - In the present study we have used FISH to analyze the incidence of trisomy 8 in acute leukemias following either a primary myeloproliferative disorder (MPD) or a myelodysplastic syndrome (MDS) and correlated it with both the immunophenotype and the cell-cycle distribution of the leukemic blast cells. Six of the 21 (28%) acute leukemias studied displayed trisomy 8 by FISH. The number of trisomic cells in these cases ranged from 20 to 84%, with a mean of 46 +/- 24%. Trisomy 8 was associated with a homogeneous population of leukemic cells, phenotypically characterized by CD34+/HLADR+/CD13+/CD33+/CD11b-/ CD15-/CD14-. No significant differences were observed on the proliferative rate of cases with trisomy 8, as compared with blast cells from the remaining patients. Overall, our findings suggest that in acute leukemias secondary to MPD or MDS, trisomy 8 is associated with a blockade of myeloid maturation at an early step of the differentiation process. PMID- 9200993 TI - Recombinant human granulocyte colony-stimulating factor increases circulating CD34-postive cells in patients with AIDS. AB - In a gene therapy-based treatment of AIDS, it would be desirable to have as many transduced target cells as possible. A limiting factor is the number of target cells. In this study, we investigated whether it was possible to increase the absolute number of one possible target cell, i.e., the circulating hematopoietic progenitor cells (CD34 cells) in patients with AIDS, using the recombinant human granulocyte colony-stimulating factor (G-CSF). Eight patients with AIDS were treated with G-CSF for neutropenia (< 1.0 x 10(9)/l). Treatment consisted of daily subcutaneous injections with 300 micrograms G-CSF for 3-5 consecutive days. Within 5 days of initiation of G-CSF therapy, an increase in the absolute neutrophil count (ANC) was seen in all patients. There was a median increase in ANC from 0.4 to 3.4 x 10(9)/l. In addition, G-CSF treatment significantly increased the absolute number of CD34 cells. The median increase in CD34 cells was from 0.8 to 2.2 x 10(6)/l. Finally, using a highly sensitive HIV-1 RNA PCR, we found that treatment of AIDS patients with G-CSF did not lead to enhanced HIV replication. These observations indicate that G-CSF may be used to mobilize CD34 cells in patients with AIDS, e.g., for a gene therapy protocol. PMID- 9200994 TI - Risk factors for capillary leakage syndrome after bone marrow transplantation. AB - Age, hematopoietic growth factors, cyclosporin A, mode of bone marrow transplantation (BMT) (autologous, allogeneic-related, unrelated), and underlying disease were assessed as potential risk factors for capillary leakage syndrome (CLS) in 96 patients after BMT. CLS was defined as unexplained weight gain of > 3% within 24 h and nonresponsiveness to furosemide. CLS occurred in 9/21 patients after unrelated compared with 2/33 after allogeneic-related BMT (p = 0.0017) for hematopoietic disorders (n = 54) and in 6/7 patients after allogeneic-related compared with 3/35 after autologous BMT (p = 0.001) for solid tumors (n = 42). Hematopoietic growth factors and cyclosporin A were no significant risk factors on their own. We conclude that unrelated BMTs or high-intensity conditioning regimens used in combination with allogeneic-related BMT are the main risk factors for CLS. PMID- 9200995 TI - G-CSF and cyclosporin induce an increase of normal cells in hypoplastic paroxysmal nocturnal hemoglobinuria. AB - Four paroxysmal nocturnal hemoglobinuria (PNH) patients with severe thrombocytopenia, hemolytic anemia and neutropenia were treated using a combination of filgrastim (G-CSF) and cyclosporin. In all patients a trilineage response of hematopoiesis was achieved. In addition, the proportion of glycosyl phosphatidylinositol (GPI)-deficient granulocytes decreased. All patients mobilized CD34+ hematopoietic progenitors into peripheral blood after starting treatment with G-CSF. The majority of early progenitors (CD34+ CD38-) after mobilization into peripheral blood was found to be unaffected by the GPI anchoring defect. No patient developed leukemia while under therapy. We conclude from these data that the combination of G-CSF and cyclosporin represents an efficient option for the treatment of hypoplastic PNH. PMID- 9200996 TI - Preclinical evaluation of biotin labeling for red cell survival testing. AB - Biotin labeling of red cells was tested in dogs as a preclinical study for cell survival. Red cells were labeled with either spacered Biotin-X-NHS (BxNHS) or water-soluble biotin compounds. After reinfusion, biotinylated red cells were detected in small blood samples (5 microliters) with flow cytometry. Improved BxNHS labeling allows an easy detection of positive red cells for almost 100 days, whereas labeling with watersoluble compounds-despite strong labeling during the first days-results in a decrease of label, which prevented a discrimation between labeled and negative cells after about 4 weeks. When biotin labeling of red cells was compared with 51Cr labeling, slopes of red cell survival were quite similar after the latter were corrected for elution. Survival slopes were linear, and the mean survival time was t = 93 d. In two blood-donor dogs the slopes of red cell survival where log linear and the mean survival time was t = 45 d. In conclusion, BxNHS, but not the water-soluble biotin compounds, is a good nonradioactive, nontoxic alternative for red cell survival studies. No health hazards are to be expected from the very low dose of Dimethylformamide, which is used as a solvent for biotin-x-NHS. PMID- 9200997 TI - Specificities of platelet autoantibodies in patients with lupus anticoagulants in primary antiphospholipid syndrome. AB - We have studied target platelet antigens in 22 patients with lupus anticoagulants and a primary antiphospholipid syndrome in order to determine whether any specificities of platelet autoantibodies are correlated with thromboembolism, and if these antibodies cross-react with phospholipids, which would suggest their role in the development of thromboembolic disease. Platelet counts were median 203 x 10(9)/l, range 100-298 x 10(9)/l. Platelet antibodies were found in six thrombocytopenic patients and in further nine patients. All these 15 patients had antibodies against GPIIb/ IIIa, five patients against GPIb/IX, and six patients against GPIV. Anti-GPIb/IX and -GPIV occurred only in combination with anti GPIIb/IIIa antibodies. There was no correlation between the presence of detectable platelet antibodies or any of their glycoprotein specificity and thrombocytopenia or the history of a thromboembolic disease. Eluates from platelets contained only GPIIb/IIIa reactivities, but neither anti-GPIb/IX nor anti-GPIV. None of the eluates contained lupus anticoagulant activity. In one case, the platelet eluates contained anti-GPIIb/IIIa and anticardiolipin IgG antibodies. These results suggest that in patients with a primary antiphospholipid syndrome the presence of platelet autoantibodies neither indicate a risk for thromboembolic disorder nor have lupus anticoagulant activity. PMID- 9200998 TI - Chronic lymphocytic leukemia with two cellular populations: a biphenotypic or biclonal disease. AB - A case of CLL with two different cellular populations is reported. A 50-year-old man was evaluated for persistent absolute lymphocytosis. A peripheral blood smear revealed numerous small lymphocytes (83% of white blood cells counted). Frequent Grumpecht shadows were present, too. On bone marrow aspirate smears lymphocytes comprised 85% of the total cells counted, and the bone marrow biopsy showed a mixed nodular-interstitial infiltration pattern. The immunophenotypic study showed two different leukemic populations. The first one (comprising 79% leukemic cells) was CD5+, CD19+, CD10-, CD20+, CD18-, CD22-, CD23+ +, lambda dim, and FMC7 . The second population (comprising 21% leukemic cells) was CD5+, CD19+, CD10-, CD20+, CD18+, CD22+, CD23+, lambda+ +, and FMC7+. Gene rearrangement studies detected the germline and one rearranged band in Jk blot with each restriction endonuclease. In the Jh blot the germline and two rearranged bands were detected with EcoRI and BamHI and three rearranged bands with HindIII. The JBI/JBII blot detected only the germline band. The detection of three rearranged bands was interpreted as evidence of the presence of at least two monoclonal populations of cells with the same light chain restriction. PMID- 9200999 TI - BCL6 rearrangement in a patient with mantle cell lymphoma. AB - We describe a patient with mantle cell lymphoma (MCL) associated with BCL6 gene rearrangement. MCL is a distinct subtype of non-Hodgkin's lymphoma characterized by CD5+, CD10-, CD20+, t(11;14)(q13;q32) and PRAD1/cyclin D1 overexpression. Although rearrangement of the BCL6 gene is the most frequent genetic change among diffuse lymphomas and some follicular lymphomas this is the first report of a patient with MCL associated with BCL6 rearrangement. PMID- 9201000 TI - Is increased renin activity as a paraneoplastic phenomenon in acute leukemia a protective mechanism against hyperkalemia? PMID- 9201001 TI - Stain therapy and reduced incidence of stroke. Implications of cholesterol lowering therapy for cerebrovascular disease. PMID- 9201002 TI - System, supervision, standards, and the 'epidemic' of negligent medical errors. PMID- 9201003 TI - The management of diabetes insipidus in adults. AB - The management of patients with diabetes insipidus can be confusing because of the disorder's variable pathophysiology, the numerous medications used, and the possible complications related to their use. Nevertheless, the primary care physician, rather than the subspecialist, will increasingly be called on to manage patients with such relatively uncommon conditions in the future. If a few basic facts and principles are kept in mind, the care of most patients with diabetes insipidus can be successful. A comprehensive, practical review of the short- and long-term therapy for patients with diabetes insipidus, including central diabetes insipidus, nephrogenic diabetes insipidus, and the "excessive vasopressinase syndrome," is presented. The use of single and multidrug regimens, and of the newly marketed oral formulation of desmopressin acetate, is described for common clinical settings. PMID- 9201004 TI - Reductase inhibitor monotherapy and stroke prevention. AB - BACKGROUND: Epidemiologic evidence and meta-analyses of data from early clinical trials suggest that lowering the levels of cholesterol does not reduce the events of stroke. These analyses have not included more recent clinical trials using reductase inhibitors. OBJECTIVE: To conduct a meta-analysis of the effect of reducing cholesterol levels on stroke in all reported clinical trials of primary (n = 4) and secondary (n = 8) prevention of coronary heart disease that used reductase inhibitor monotherapy and provided information on incident stroke. RESULTS: Analysis of combined data from primary and secondary prevention trials showed a highly statistically significant reduction of stroke associated with the use of reductase inhibitor monotherapy (27% reduction in stroke; P = .001). Analysis of secondary prevention trials alone disclosed a similar statistically significant effect (32% reduction in stroke; P = .001). A smaller nonsignificant reduction in stroke was noted in the primary prevention trials (15% reduction in stroke; P = .48). CONCLUSIONS: Reductase inhibitors now in use for lowering cholesterol levels are more potent and have fewer side effects than the cholesterol-lowering agents previously available. They appear to reduce stroke, most notably in patients with prevalent coronary artery disease, which may be partly due to the effects of lowering the levels of cholesterol on the progression and plaque stability of extracranial carotid atherosclerosis or the marked reduction of incident coronary heart disease associated with treatment. PMID- 9201005 TI - Physicians counseling smokers. A population-based survey of patients' perceptions of health care provider-delivered smoking cessation interventions. AB - OBJECTIVE: To examine associations between sociodemographic and psychological characteristics of smokers and delivery of 5 types of smoking cessation counseling interventions by physicians and office staff. METHODS: We used a telephone survey of a population-based sample of adult cigarette smokers (N = 3037) who saw a physician in the last year. Primary outcomes included patients' report of whether a physician or other health care provider (1) talked about smoking, (2) advised them to quit, (3) offered help to quit, (4) arranged a follow-up contact, and (5) prescribed nicotine gum or other medication. RESULTS: Fifty-one percent of smokers were talked to about their smoking; 45.5% were advised to quit; 14.9% were offered help; 3% had a follow-up appointment arranged; and 8.5% were prescribed medication. In multivariate analyses, the most consistent predictors of receipt of almost all counseling behaviors were medical setting (private physician's office only > care in other settings), health status (fair or poor > good, very good, or excellent), more years of education, greater number of cigarettes smoked per day, stage of readiness to quit smoking (preparation > precontemplation), and greater reported benefits of smoking. CONCLUSIONS: Physicians and other health care providers are not meeting the standards of smoking intervention outlined by the National Cancer Institute and the Agency for Health Care Policy and Research. Health care providers who intervene only with those patients who are ready to quit smoking are missing opportunities to provide effective smoking interventions to the majority of their patients. Interventions are also less likely to be provided to healthier and lighter smokers. PMID- 9201006 TI - Organizing pneumonia. Features and prognosis of cryptogenic, secondary, and focal variants. AB - BACKGROUND: Organizing pneumonia (OP) is a non-specific response to many types of lung injury. Clinicians frequently encounter pathology reports of OP in patients with no underlying condition (cryptogenic OP, also known as BOOP or bronchiolitis obliterans OP) or in association with drugs or nonpulmonary disease. The goals of this study are to describe the clinical course and outcomes in patients with 3 clinical variants of OP. METHODS: A retrospective study of patients with OP seen at the Mayo Clinic, Rochester, Minn, from January 1, 1984, through June 30, 1994, was conducted. Initial features were obtained from medical records. Chest radiographs and pathology specimens were reviewed for this study. Resolution, relapse, and survival were obtained from medical records and a follow-up patient questionnaire. RESULTS: Seventy-four patients had pathologically confirmed OP. Organizing pneumonia was classified into 3 clinical groups: symptomatic cryptogenic OP; symptomatic OP related to underlying hematologic malignant neoplasm, collagen vascular disease, or drugs (secondary OP); and asymptomatic OP presenting as a focal nodule (focal OP). Thirty-seven patients (50%) had cryptogenic OP and 27 patients (36%) had secondary OP. No difference was found between cryptogenic and secondary OP in type or severity of symptoms, signs, laboratory and pulmonary function tests, or radiologic or pathologic findings. Corticosteroids were given at a similar initial dose (prednisone, about 50 mg/d). Resolution of symptoms was more frequent in patients with cryptogenic OP than those with secondary OP. Relapse was infrequent in both of these groups. Five year survival was higher in patients with cryptogenic OP (73%) than in secondary OP (44%), and respiratory-related deaths were more frequent in patients with secondary OP. Organizing pneumonia was an asymptomatic focal rounded opacity in 10 patients (14%), most often detected on chest radiograph and diagnosed on lung biopsy done for suspicion of lung cancer. Patients with focal OP required no treatment and had no relapse or respiratory-related deaths. CONCLUSIONS: Clinical classification of OP is useful to predict clinical course and outcome. Cryptogenic OP most often was a symptomatic bilateral lung process that had an overall favorable prognosis with prolonged corticosteroid therapy. Patients with secondary OP had a high mortality rate when the disease was associated with predisposing conditions or drugs. Patients with asymptomatic focal OP had an excellent prognosis. PMID- 9201008 TI - Use of postmenopausal hormone replacement therapy by African American women. The importance of physician discussion. AB - BACKGROUND: Although nationally the use of hormone replacement therapy (HRT) has increased dramatically in the last decade, little is known about its use by disadvantaged, minority women or the role that physician discussion plays in determining its use. METHODS: In 1994, we surveyed a total of 328 predominantly indigent, African American women (refusals, 22) who attended public hospital medical continuity of care clinics staffed by internal medicine house officers. RESULTS: Of the 328 women who completed the survey, the mean age was 63 years, 302 women (92%) were African American, and 286 (87.3%) had yearly incomes of less than $10000. Of the 328 women, 52 (15.6%) were receiving HRT at the time of the survey, varying from 22 (28.2%) of 78 women aged 50 to 59 years to 1 (3%) of 33 women older than 80 years (P = .006). In a logistic regression model adjusting for age and ethnicity, women who had previously undergone a hysterectomy were significantly more likely to use HRT, with an odds ratio of 2.76 (95% confidence interval, 1.44-5.30; P = .002). The levels of education and income and the history of myocardial infarction were not significantly associated with the use of HRT (P > .20). Although all women who were currently receiving HRT recalled discussing HRT with their physicians, of the 276 women who were not receiving HRT, only 62 (22%) recalled any such discussion (P < .001). CONCLUSIONS: In poor, African American women who reside in the inner city, younger age, undergoing a hysterectomy, and physician discussion of HRT are significantly associated with current use of HRT. Physician discussion of HRT may need greater emphasis in internal medicine training programs. PMID- 9201007 TI - Duration of estrogen replacement therapy in relation to the risk of incident myocardial infarction in postmenopausal women. AB - BACKGROUND: There is little information about whether an increasing duration of estrogen replacement therapy is associated with a declining risk for myocardial infarction in postmenopausal women. OBJECTIVE: To conduct a population-based, case-control study among enrollees of the Group Health Cooperative (GHC) of Puget Sound, Seattle, Wash. SUBJECTS AND METHODS: Case subjects were all post menopausal women who were enrolled in the GHC with an incident fatal or nonfatal myocardial infarction from July 1986 through December 1993. Control subjects were a stratified random sample of postmenopausal women who were enrolled in the GHC without myocardial infarction and matched to case subjects by age and calendar year. We reviewed the medical records of the 850 case subjects and 1974 control subjects and conducted telephone interviews with consenting survivors. Use of estrogen or estrogen and progestin was assessed using GHC's computerized pharmacy database. RESULTS: Among women who were currently using estrogen, a longer duration of use was inversely associated with a risk for myocardial infarction after adjustment for age, year of identification, diabetes mellitus, angina, and smoking. For categories of increasing duration of estrogen use (never, > 0-< 1.8 years, 1.8-< 4.2 years, 4.2-< 8.2 years, and > or = 8.2 years), the odds ratios for myocardial infarction were 1.00 (reference), 0.91, 0.70, 0.65, and 0.55 (for trend among the current users, P = .05). Among women who had used estrogen in the past, there was no evidence of decreasing risk with increasing duration of estrogen use. CONCLUSION: In this study, a long duration of hormone replacement therapy among women currently using estrogen was associated with a reduced risk for first myocardial infarction. PMID- 9201009 TI - Outcome of hypertension management in Asian Americans. AB - BACKGROUND: Ethnic and/or racial differences in drug response to antihypertensive agents have been recognized, yet the prescribing practices and the information on efficacy of various agents rely mainly on the response of whites to drugs. OBJECTIVES: To assess the management of hypertension in Asian Americans and to compare it with an age- and sex-matched group of white patients with hypertension. METHODS: The patients' medical records were used as the primary source of information for the data collection. The observational period was a 12 month window and included 200 patients of Asian origin with hypertension and 196 white patients with hypertension whose medical records were randomly selected. RESULTS: The study describes the pattern of use of antihypertensive agents and the differences in response to antihypertensive agents between Asian Americans and whites. The preferred antihypertensive agents in both Asian and white patients included monotherapy with either calcium channel blockers or angiotensin converting enzyme inhibitors. However, medication changes, dose reduction, and the experience of side effects were all significantly more frequently recorded in Asian patients than in white patients (P < .001, P < .008, and P < .002, respectively). CONCLUSIONS: These findings are supportive of some previous reports on ethnic differences in drug response to antihypertensive agents. The findings also point to the need for further prospective studies on the outcome of hypertension management in Asian American patients. PMID- 9201010 TI - Choice of treatment improves quality of life. A study on patients undergoing dialysis. AB - BACKGROUND: Quality of life (QOL) is an important measure of the success of medicine. Choice of treatment is an important variable influencing QOL. We studied QOL in patients undergoing treatment for end-stage renal failure. Until June 1993 our patients needing dialysis could freely choose continuous ambulatory peritoneal dialysis (CAPD); however, since that time most patients have been forced to undergo CAPD because the hemodialysis program is full. METHODS: We compared QOL in patients accepted before or after June 1993. Forty-five patients undergoing CAPD were studied during the period of choice compared with 44 who had no choice. Quality of life was studied by Bradburn Affect Scale, Mental Health Scale, Campbell Life Satisfaction, Perceived Health, Karnofsky Scale, Activity Scale, Physical Symptoms Scale, and desire for treatment change. RESULTS: The patients undergoing CAPD in the no-choice group had a lower score than the choice population in 4 of the 7 QOL scales. The Mental Health Scale mean score was 18.4 compared with 15.5, and the patients ranking highest on the Mental Health Scale decreased from 33% to 18%, while those ranking lowest increased 7-fold from 2% to 14% comparing choice with no-choice group. The Bradburn Affect Scale score was +0.7 in the choice group compared with -0.3 in the no-choice group. There were no differences in age, sex, race, or treatment that explained the difference. Influence of other time-related factors is unlikely as there were no similar lower scores with time in the QOL reported by patients in the in-center or assisted self-care hemodialysis or transplant groups. CONCLUSIONS: Once the freedom of choice of treatment is gone from the patients undergoing CAPD their psychological QOL deteriorates. PMID- 9201011 TI - Postcoital vaginal bleeding as a risk factor for transmission of the human immunodeficiency virus in a heterosexual partner study in Brazil. Rio de Janeiro Heterosexual Study Group. AB - BACKGROUND: Several risk factors for male-to-female human immunodeficiency virus (HIV) transmission are well established, but few studies have examined the role of postcoital vaginal bleeding. METHODS: Couples recruited from AIDS centers in Rio de Janeiro, Brazil, were interviewed for risk factors and had blood collected for examinations. Eligibility criteria included confirmed HIV positivity for the male partner, aged 18 years of older, heterosexual contact in the past year, and no other risk factor for female partners except sexual contact with the HIV infected male partner. Logistic regression was used to assess the association between HIV serostatus and risk factors in the female partners. RESULTS: The prevalence of HIV infection was 47.6% among the 418 women available for analysis. The following factors were independently associated with HIV infection; anal sex (odds ratio [OR], 3.06), condom use during vaginal sex sometimes (OR, 1.42) and rarely to never (OR, 2.00) compared with always, frequency of sexual contacts (> 100 in the previous year) (OR, 1.71), HIV-infected male partners with symptoms of acquired immunodeficiency syndrome (OR, 1.70), and postcoital vaginal bleeding (OR, 1.89). The association of postcoital bleeding and HIV infection was more pronounced among women who did not engage in anal sex. CONCLUSIONS: The results support previous studies showing that advanced HIV infection in the male partner, anal sex, inconsistent condom use, and frequent sex are associated with HIV infection among the female partners of HIV-infected men. Postcoital vaginal bleeding was also identified as a risk factor for infection. In addition to other established preventive measures, a recommendation for seeking diagnosis and treatment of sexually transmitted diseases that are associated with postcoital bleeding and using water-soluble lubricants during sex to minimize trauma seems prudent. PMID- 9201012 TI - Smoking cessation after surgery. A randomized trial. AB - BACKGROUND: Cigarette smoking is the greatest cause of preventable mortality in the United States. Because most smokers would like to quit and most hospitals are smoke free, surgical admissions represent a window of opportunity for tobacco cessation interventions. METHODS: A total of 324 patients (98% men), aged 25 to 82 years, who were current smokers and who underwent noncardiac surgery were enrolled in a randomized controlled trial at the Veterans Affairs Medical Center, San Francisco, Calif. One hundred sixty-eight participants (52%) received a multicomponent intervention designed to increase self-efficacy and coping skills that included face-to-face in-hospital counseling, viewing a smoking cessation videotape, self-help literature, nicotine replacement therapy, and 3 months of telephone follow-up. One hundred fifty-six participants (48%) received self-help literature and brief counseling lasting 10 minutes. Serum or saliva cotinine levels were measured to confirm self-reported smoking cessation. RESULTS: At 12 months of follow-up, the self-reported quit rate was 27% among the intervention group and 13% among the comparison group (relative risk, 2.1; 95% confidence interval, 1.2-3.5; P < .01). Based on biochemical confirmation, 15% of the intervention group, compared with 8% of the comparison group, quit smoking at 12 months (relative risk, 2.0; 95% confidence interval, 1.0-3.9; P = .04). CONCLUSIONS: A smoking cessation intervention targeted at smokers hospitalized for noncardiac surgery can increase long-term quit rates. Surgical hospitalizations provide an opportunity to reach smokers who want to quit smoking. PMID- 9201014 TI - Vulnerability to warfarin: could undernutrition be a predictor? PMID- 9201013 TI - Association of gender and survival in patients with acute myocardial infarction. AB - BACKGROUND: During the last 5 years, many studies have produced conflicting results concerning the survival of women hospitalized with acute myocardial infarction (AMI). OBJECTIVE: To determine if gender is associated with hospital mortality and long-term survival in individuals with AMI. METHODS: This prospective study included 4255 consecutive women (34%) and 8076 (66%) men who developed AMI in 19 Seattle, Wash, area hospitals between January 1988 and June 1994. Key information was abstracted from hospital records and entered in the Myocardial Infarction Triage and Intervention registry database. In addition, data concerning survival and rehospitalization were obtained from the state of Washington and linked to the Myocardial Infarction Triage and Intervention registry. RESULTS: In comparison with men, women were 8 years older, more often had history of congestive heart failure, hypertension, or diabetes mellitus, and less often had history of myocardial infarction or coronary surgery. During hospitalization, women were less likely to undergo coronary angiography, thrombolytic therapy, coronary angioplasty, or bypass surgery. After adjustment for covariates, women were 20% more likely to die in the hospital (odds ratio, 1.22; 95% confidence interval, 1.06-1.39), yet long-term survival was similar in the 2 groups (hazard ratio, 0.97; 95% confidence interval, 0.90-1.05). The use of thrombolytic therapy or revascularization during the index hospitalization did not change the association between gender and survival. CONCLUSIONS: All things being equal, women with AMI were more likely to die in the hospital, yet survival after hospital discharge did not differ according to gender. Appropriate treatment to reduce hospital mortality in women is needed. PMID- 9201015 TI - Thyrotropin testing in recent-onset atrial fibrillation. PMID- 9201016 TI - Familial pheochromocytoma due to mutant von Hippel-Lindau disease gene. PMID- 9201017 TI - Tuberculous pericarditis in a woman infected with human immunodeficiency virus type 1, long-term nonprogressive. PMID- 9201018 TI - Acute promyelocytic leukemia following leiomyosarcoma of the jejunum. PMID- 9201019 TI - Colonic histoplasmosis associated with acquired immunodeficiency syndrome. PMID- 9201020 TI - Hospital readmissions for elderly patients with congestive heart failure. PMID- 9201021 TI - Cyclic strain-induced monocyte chemotactic protein-1 gene expression in endothelial cells involves reactive oxygen species activation of activator protein 1. AB - Endothelial cells (ECs) are constantly exposed to blood pressure-induced mechanical strain. We have previously demonstrated that cyclic strain can induce gene expression of monocyte chemotactic protein-1 (MCP-1). The molecular mechanisms of gene induction by strain, however, remain unclear. Recent evidence indicates that intracellular reactive oxygen species (ROS) can act as a second messenger for signal transduction and thus affect gene expression. The potential role of ROS in strain-induced MCP-1 expression was investigated. ECs under cyclic strain induced a sustained elevated production of intracellular superoxide. ECs under strain or pretreated with either H2O2 or xanthine oxidase/hypoxanthine induced MCP-1 expression. Strain- or oxidant-induced MCP-1 mRNA levels could be inhibited by treating ECs with catalase or antioxidant N-acetyl-cysteine (NAC). Functional analysis of MCP-1 promoter and site-specific mutations indicates that the proximal tissue plasminogen activator-responsive element (TRE) in the -60-bp promoter region is sufficient for strain or H2O2 inducibility. Electrophoretic mobility shift assays demonstrated an increase of nuclear proteins binding to TRE sequences from ECs subsequent to strain or H2O2 treatment. NAC or catalase pretreatment of ECs inhibited the strain- or H2O2-induced AP-1 binding. These results clearly indicate that cyclic strain inducibility of MCP-1 in ECs uses the interaction of AP-1 proteins with TRE sites via the elevation of intracellular ROS levels in strained ECs. These findings emphasize the importance of intracellular ROS in the modulation of hemodynamic force-induced gene expression in vascular ECs. PMID- 9201022 TI - Monocytes harboring cytomegalovirus: interactions with endothelial cells, smooth muscle cells, and oxidized low-density lipoprotein. Possible mechanisms for activating virus delivered by monocytes to sites of vascular injury. AB - Cytomegalovirus (CMV) infection and its periodic reactivation from latency may contribute to atherogenesis and restenosis. It is unknown how CMV is delivered to the vessel wall and is reactivated. We examined the following hypothesis: CMV, present in monocytes recruited to sites of vascular injury, is activated by endothelial cell (EC) or smooth muscle cell (SMC) contact and by oxidized low density lipoproteins (oxLDLs). The CMV major immediate-early promoter (MIEP) controls immediate-early (IE) gene expression, and thereby viral replication. To determine whether elements of the vessel wall can activate CMV present in monocytes, we transiently transfected the promonocytic cell line HL-60 with a chloramphenicol acetyltransferase reporter gene construct driven by MIEP. MIEP activity increased 1.7 +/- 0.5-fold (P = .02) when the transfected HL-60 cells were cocultured with ECs, 4.5 +/- 1.5-fold when cocultured with SMCs (P = .03), and 2.0 +/- 0.5-fold (P = .01) when exposed to oxLDL. The combination of oxLDL and EC coculture increased MIEP activity over 7-fold. We also found that freshly isolated human monocytes, infected with endothelium-passaged CMV, were capable of transmitting infectious virus to cocultured ECs or SMCs. CMV-related progression of atherosclerosis or restenosis may, at least in part, involve monocyte delivery of the virus to the site of vascular injury, where the vascular milieu, ie, contact with ECs, SMCs, and oxLDL, can contribute to viral reactivation and/or replication by enhancing CMV IE gene expression. The virus may then infect neighboring ECs or SMCs, initiating a cascade of events predisposing to the development of atherogenesis-related processes. PMID- 9201023 TI - Diverse effects of heparin on mitogen-activated protein kinase-dependent signal transduction in vascular smooth muscle cells. AB - Proliferation of vascular smooth muscle cells (SMCs) is implicated in pathological events, including atherosclerosis and intimal hyperplasia following angioplasty. The glycosaminoglycan heparin is a growth inhibitor of SMCs in vitro and in vivo. The underlying mechanism, however, is still poorly understood. In the present study, we report that heparin inhibited the activation of the mitogen activated protein kinase (MAPK) in baboon SMCs by serum but not by platelet derived growth factor (PDGF). When fibroblast growth factor was used, heparin had a stimulatory effect on MAPK. The only MAPK-activating kinase found in SMCs was MAPK kinase (MAPKK)-1, although MAPKK-2 was present in comparable amounts. Activation of MAPKK-1 and DNA synthesis were affected by heparin in a similar fashion. Heparin does not appear to exert its effects through members of the protein kinase C family, which are downregulated by phorbol esters, because it was still capable of inhibiting MAPK/MAPKK-1 stimulation by FCS in phorbol ester pretreated cells. The present findings support the conclusions that the effects of heparin depend on the nature of the mitogen and that heparin inhibits SMC proliferation by preventing activation of MAPKK-1. PMID- 9201024 TI - Induction of vascular endothelial growth factor in balloon-injured baboon arteries. A novel role for reactive oxygen species in atherosclerosis. AB - Neovascularization is a hallmark of neointimal formation in atherosclerotic plaques and restenotic lesions. Vascular endothelial growth factor (VEGF) promotes neovascular growth, whereas oxidative stress is a potent factor in vascular cell proliferation. To investigate the mechanisms of neovascular formation, we treated human and rat vascular smooth muscle cells (VSMCs) with H2O2. Northern blot analysis demonstrated a dose- and time-dependent increase in VEGF mRNA, with a maximum of 4-fold at 3 hours (200 mumol/L). As determined by immunoblotting and enzyme-linked immunosorbent assay, VEGF protein expression and secretion were similarly increased. Human umbilical vein endothelial cells were treated with conditioned medium from VSMCs incubated with 200 mumol/L H2O2. DNA synthesis, measured by thymidine incorporation, was increased 4-fold compared with control, an effect that was blocked by a neutralizing anti-VEGF antibody. The lipid peroxidation product 4-hydroxynonenal (1 mumol/L), an endogenous reactive oxygen species present in human atherosclerotic lesions, also increased VEGF secretion in VSMCs in a similar time-dependent fashion. Immunohistochemical staining and in situ hybridization of aortic sections from balloon-injured baboons demonstrated increased VEGF expression in discrete areas of the neointima and media compared with control sections, and expression correlated with the generation of 4-hydroxynonenal. Regulators of VEGF expression, such as reactive oxygen species, may enhance neovascularization of atherosclerotic and restenotic arteries. PMID- 9201025 TI - Pulmonary vasoconstriction and hypertension in mice with targeted disruption of the endothelial nitric oxide synthase (NOS 3) gene. AB - NO, synthesized in endothelial cells by endothelial NO synthase (NOS 3), is believed to be an important endogenous pulmonary vasodilator substance that contributes to the normal low pulmonary vascular resistance. To selectively investigate the role of NOS 3 in the pulmonary circulation, mice with targeted disruption of the NOS 3 gene were studied. Pulmonary hemodynamics were studied by measuring pulmonary artery pressure, left ventricular end-diastolic pressure, and lower thoracic aortic flow by using a novel open-chest technique. Transient partial occlusion of the inferior vena cava was used to assess the pulmonary artery pressure-flow relationship. Tension developed by isolated pulmonary artery segments after acetylcholine stimulation was measured in vitro. The histological appearance of NOS 3-deficient and wild-type murine lungs was compared. NOS 3 deficient mice (n = 27), when compared with wild-type mice (n = 32), had pulmonary hypertension (pulmonary artery pressure, 19.0 +/- 0.8 versus 16.4 +/- 0.6 mm Hg [mean +/- SE]; P < .05) that was due to an increased total pulmonary resistance (62 +/- 6 versus 33 +/- 2 mm Hg.min.g.mL-1; P < .001). In vitro, acetylcholine induced vasodilation in the main pulmonary arteries of wild-type but not NOS 3-deficient mice. The morphology of the lungs of NOS 3-deficient mice did not differ from that of wild-type mice. We conclude that NOS 3 is a key enzyme responsible for providing basal pulmonary NO release. Congenital NOS 3 deficiency produces mild pulmonary hypertension in mice. PMID- 9201026 TI - Evidence that late preconditioning against myocardial stunning in conscious rabbits is triggered by the generation of nitric oxide. AB - Recent studies in conscious pigs and rabbits have demonstrated that a series of brief coronary occlusions renders the heart relatively resistant to myocardial "stunning" 24 hours later (late preconditioning [PC] against stunning). The mechanism of this powerful cardioprotective response is unknown. The goal of the present study was to test the hypothesis that the development of late PC against stunning is triggered by increased generation of NO during the first ischemic challenge. Conscious rabbits underwent a sequence of six 4-minute coronary occlusion/4-minute reperfusion cycles for 3 consecutive days (days 1, 2, and 3). On day 1, rabbits received either an intravenous infusion of the NO synthase inhibitor NG-nitro-L-arginine (L-NA, 13 mg/kg before the first occlusion) (group II, n = 10) or vehicle (group I [control], n = 10). In the control group, on day 1 systolic wall thickening (WTh) in the ischemic/reperfused region remained significantly depressed for 4 hours after the sixth reperfusion, indicating myocardial stunning. On days 2 and 3, however, the recovery of WTh improved markedly, so that the total deficit of WTh decreased by 60% on day 2 and 55% on day 3 compared with day 1 (P < .01). In the L-NA-treated group, the total deficit of WTh on day 1 was similar to that observed in the control group. On day 2, however, the total deficit of WTh was not significantly different from that observed on day 1 and was 132% greater than that observed in control rabbits on day 2 (P < .01). On day 3, the total deficit of WTh was 66% less than that noted on day 2 (P < .01). Thus, in L-NA-treated rabbits the sequence of six coronary occlusions and reperfusions performed on day 1 failed to precondition against stunning on day 2, but the same sequence performed on day 2 did precondition against stunning on day 3. Another group of rabbits (group III, n = 6) received L NA on day 1 in the absence of ischemia and was subjected to the occlusion/ reperfusion sequence on days 2 and 3. In these animals, the total deficit of WTh on day 2 did not differ from that observed in control rabbits on day 1, indicating that administration of L-NA did not exacerbate the severity of myocardial stunning 24 hours later; therefore, the absence of late PC against stunning on day 2 in group II cannot be ascribed to a delayed deleterious action of L-NA on WTh. In conclusion, these results demonstrate that the NO synthase inhibitor L-NA completely blocks the development of late PC against myocardial stunning in conscious rabbits, indicating that NO generated as a result of the PC ischemia triggers the development of the cardioprotective response observed 24 hours later. NO is known to exert numerous biological actions resulting in rapid but transient physiological responses. The present observations support a novel pathophysiological paradigm in which NO also plays a key role in the delayed myocardial adaptations to ischemic stress, acting as a signaling step in the transduction pathway that leads to increased resistance to subsequent ischemic injury. PMID- 9201027 TI - Delayed enhanced nitric oxide-mediated coronary vasodilation following brief ischemia and prolonged reperfusion in conscious dogs. AB - The goal of this study was to determine both the early and delayed effects of a brief (10-minute) coronary artery occlusion (CAO) and prolonged (5-day) reperfusion (CAR) on coronary endothelial function. Fourteen mongrel dogs were chronically instrumented to measure aortic and left ventricular pressures, wall thickness, and left circumflex coronary blood flow (CBF). Before CAO and during CAR, coronary vascular reactivity was investigated by 15-second CAO and subsequent reactive hyperemia (RH) and by the selective intracoronary infusion of acetylcholine (ACh, 10 micrograms/min) and bradykinin (BK, 2.5 micrograms/min), endothelium-dependent vasodilators, and sodium nitroprusside (SNP, 40 micrograms/min), an endothelium-independent vasodilator. CBF responses to ACh and BK began to increase after 6 hours of CAR, reached a peak after 1 to 2 days of CAR, and then subsided over the subsequent 4 days. After 1 day of CAR, compared with before CAO, enhanced CBF responses (P < .05), associated with increased coronary sinus oxygen content, were observed for-ACh (+66 +/- 20%), BK (+74 +/- 24%), and RH (+24 +/- 5%) but not SNP (-2 +/- 10%). Production of NO metabolites (nitrate and nitrite), measured as their coronary arteriovenous differencexCBF, was significantly increased after 1 to 2 days of CAR, both at baseline (153 +/- 56%) and during BK infusion (220 +/- 76%) (P < .05). Holding CBF at pre-CAO levels during the initial CAR period did not attenuate the delayed enhanced endothelial vasodilation to ACh and BK. However, NO blockade with intracoronary NG-nitro-L-arginine blocked the enhanced coronary vasodilation to ACh and BK. Thus, in contrast to previous studies, these data indicate that brief ischemic episodes induce delayed enhanced coronary endothelial function, which is delayed in onset and prolonged in duration. This can be explained by an upregulation of coronary vascular NO production, potentially involved in the mechanism of the delayed window of preconditioning. PMID- 9201028 TI - Nitric oxide can increase heart rate by stimulating the hyperpolarization activated inward current, I(f). AB - We investigated the chronotropic effect of increasing concentrations of sodium nitroprusside (SNP, n = 8) or 3-morpholinosydnonimine (SIN-1, n = 6) in isolated guinea pig spontaneously beating sinoatrial node/atrial preparations. Low concentrations of NO donors (nanomolar to micromolar) gradually increased the beating rate, whereas high (millimolar) concentrations decreased it. The increase in rate was (1) enhanced by superoxide dismutase (50 to 100 U/mL, n = 6), (2) prevented by the guanylyl cyclase inhibitors 6-anilino-5,8-quinolinedione (5 mumol/L, n = 6) or 1H-(1,2,4)oxadiazolo(4,3-a)quinoxalin-1-one (10 mumol/L, n = 6), and (3) mimicked by 8-bromo-cGMP (n = 6) with no additional positive chronotropic effect of SIN-1 (n = 5). The response to 10 mumol/L SNP (n = 28) or 50 mumol/L SIN-1 (n = 16) was unaffected by IcaL antagonism with nifedipine (0.2 mumol/L) but was abolished after blockade of the hyperpolarization-activated inward current (I(f)) by Cs+ (2 mmol/L) or 4-(N-ethyl-N-phenylamino)-1,2-dimethyl 6-(methylamino)pyrimidinium chloride (1 mumol/L). The effect on I(f) was further evaluated in rabbit isolated patch-clamped sinoatrial node cells (n = 21), where we found that 5 mumol/L SNP or SIN-1 caused a reversible Cs(+)-sensitive increase in this current (+130% at -70 mV and +250% at -100 mV). In conclusion, NO donors can affect pacemaker activity in a concentration-dependent biphasic fashion. Our results indicate that the increase in beating rate is due to stimulation of I(f) via the NO-cGMP pathway. This may contribute to the sinus tachycardia in pathological conditions associated with an increase in myocardial production of NO. PMID- 9201029 TI - Mitochondrial response to heart rate steps in isolated rabbit heart is slowed after myocardial stunning. AB - The oxidative capacity of mitochondria isolated from myocardium is undiminished after myocardial stunning, which is remarkable because stunning affects many other cellular functions. The aim of the present study was to assess the mitochondrial oxidative response in intact rather than isolated myocardium. The mean response time of mitochondrial O2 consumption to heart rate steps (tmito) was measured before and after 15-minute ischemia or high-flow hypoxia in isolated rabbit hearts. The tmito was calculated from the time course of venous O2 tension to steps in heart rate, with corrections made for diffusion and vascular transport delay. Isovolumic hearts were perfused with Tyrode's solution at 37 degrees C. Developed left ventricular pressure at 35 minutes of reperfusion was decreased significantly to 67 +/- 3% after ischemia (mean +/- SEM, n = 8) and to 79 +/- 6% after hypoxia (n = 8) relative to the control condition (n = 8), without increased cellular creatine kinase release. Before ischemia or hypoxia, tmito was 4.3 +/- 0.3 seconds. During reperfusion after ischemia or hypoxia, the increase in tmito (by 62 +/- 10% and 64 +/- 18%, respectively) was significantly larger than that in time controls (24 +/- 12% increase). The major determinant of decreased contractility and slower mitochondrial response appeared to be O2 deprivation and/or reintroduction rather than other consequences of stopped flow. O2 consumption at a given rate-pressure product was not increased after ischemia or hypoxia, indicating undiminished cardiac contractile economy. Brief ischemia or hypoxia, resulting in stunning, was associated with a slowing of the in vivo mitochondrial oxidative response, indicating that energy transfer and/or signaling between energy-consuming sites and mitochondria is affected in stunned myocardium. PMID- 9201030 TI - Expression of a mutant (Arg92Gln) human cardiac troponin T, known to cause hypertrophic cardiomyopathy, impairs adult cardiac myocyte contractility. AB - The mechanism(s) by which mutations in sarcomeric proteins cause hypertrophic cardiomyopathy (HCM) remains unknown. A leading hypothesis proposes that mutant sarcomeric proteins impair cardiac myocyte contractility, providing an impetus for compensatory hypertrophy. To test this hypothesis, we determined the impact of expression of a mutant (Arg92Gln) human cardiac troponin T (cTnT), known to cause HCM in humans, on adult cardiac myocyte contractility. A full-length human cTnT cDNA was cloned, and the Arg92Gln mutation was induced. Recombinant adenoviruses Ad5/CMV/cTnT-N and Ad5/CMV/cTnT-Arg92Gln were generated through homologous recombination. Adult feline cardiac myocytes were infected with recombinant adenoviruses or a control viral vector (Ad5 delta E1) at a multiplicity of infection of 100. Expression levels of the full-length normal and mutant cTnT proteins were equal on Western blots. Expression of the exogenous cTnT proteins in cardiac myocytes was also shown by immunocytochemistry and immunofluorescence, and their incorporation into myofibrils was confirmed by Western blotting on myofibrillar extracts. Electron microscopy showed intact sarcomere structure in rod-shaped cardiac myocytes in all groups. Cell fractional shortening and the peak velocity of shortening were not significantly different among the groups 24 hours after transduction. However, 48 hours after transduction, both fractional shortening and the peak velocity of shortening were significantly reduced (24% [P < .001] and 26% [P < .001], respectively) in cardiac myocytes in the Ad5/CMV/cTnT-Arg92Gln compared with the Ad5/CMV/cTnT-N groups. The magnitude of the reductions was greater at 72 hours after transduction (45% and 39%, respectively; P < .001). Our results indicated that expression of the mutant (Arg92Gln) cTnT, known to cause HCM in humans, impaired intact adult cardiac myocyte contractility. Our data also show that both normal and mutant cTnT were incorporated into myofibrils. These results provide a potential mechanism by which mutations in sarcomeric proteins cause HCM. PMID- 9201031 TI - Type B atrial natriuretic peptide receptor in cardiac myocyte caveolae. AB - We have previously shown that atrial natriuretic peptide (ANP) is present in caveolae of in situ rat atrial myocytes. To investigate whether intracaveolar ANP of rat atrial myocytes exists within caveolae bound to type B ANP receptors (ANP RB, a guanylyl cyclase), we have used confocal immunofluorescence microscopy applied to primary cultures of atrial myocytes from adult rats and to freshly dissociated rat atrial myocytes (not cultured). These experimental designs tested whether atrial myocyte ANP-RB colocalizes at the plasmalemma and elsewhere in the cell with the muscle-specific isoform of the caveolar coating protein caveolin-3, and with a fraction of cellular ANP. The experiments showed that cellular caveolin-3, a fraction of cellular ANP-RB, and a fraction of cellular ANP colocalize at the plasmalemma of cultured atrial myocytes and of freshly dissociated atrial myocytes. The observations support the hypothesis that in rat atrial myocytes, intracaveolar ANP is bound to ANP-RB, a protein whose cytosolic amino acid sequences are known to encode guanylyl cyclase activity. We suggest that among the (probably multiple) effects of the cGMP thus generated in the cytoplasmic microdomain underlying atrial myocyte caveolae may be the activation of cGMP-dependent protein kinase, which would thereby inhibit plasma membrane Ca2+ channel activity and contribute to a negative inotropic effect of ANP. PMID- 9201032 TI - Positive inotropy mediated by diacylglycerol in rat ventricular myocytes. AB - Many neurohormones stimulate phospholipid hydrolysis and elevate diacylglycerol in the mammalian heart, but the physiological consequences of these intracellular events are unclear. Regulation of myocardial contraction by diacylglycerol was investigated in the present study by releasing the diacylglycerol analogue dioctanoylglycerol (diC8) within adult rat ventricular myocytes by using a light sensitive caged compound. This approach permitted us to avoid exposure of myocytes to extracellular diC8 and yet to control the amount of diC8 released into the cells. Photorelease of diC8 produced a slowly developing (half-time, 1.9 +/- 0.1 minute; n = 26) but robust (406 +/- 42%) enhancement of twitch amplitude in electrically paced myocytes (0.5 Hz, 1 mmol/L Ca2+, Ringer's solution [pH 7.4], 22 degrees C). This positive inotropic effect was dose dependent, stereospecific for the S-enantiomer of diC8, synergistically enhanced by arachidonic acid, and blocked by the protein kinase C inhibitor chelerythrine. The data provide evidence that diacylglycerol can induce a strong positive inotropic effect in mammalian ventricular muscle, possibly by activating protein kinase C. By contrast, perfusion of diC8 extracellularly onto myocytes caused a 42 +/- 2% decline in twitch amplitude, in accordance with previous reports. To account for this dependence on how diC8 is applied, we postulate that diC8 has distinct physiological actions at intracellular and extracellular sites. The peptide neurohormone endothelin-1, which elevates diacylglycerol in cardiac tissues, produced a positive inotropic effect that was similar to the response to photoreleased diC8. The diacylglycerol/protein kinase C pathway has now become a good candidate for mediator of at least a component of the positive inotropy associated with agents that stimulate phospholipid turnover in adult mammalian myocardium. PMID- 9201033 TI - Inhibitory effects of glibenclamide on cystic fibrosis transmembrane regulator, swelling-activated, and Ca(2+)-activated Cl- channels in mammalian cardiac myocytes. AB - Recent studies have provided evidence that sulfonylureas, in addition to blocking ATP-sensitive K+ (KATP) channels, also inhibit cystic fibrosis transmembrane regulator (CFTR) Cl- channels in epithelial and cardiac cells. The purpose of this study was to test whether the sulfonylurea glibenclamide might also inhibit other types of cardiac Cl- channels. Whole-cell patch-clamp techniques were used to compare the effects of glibenclamide on CFTR Cl- currents in guinea pig ventricular myocytes, swelling-activated Cl- currents in guinea pig atrial myocytes, and Ca(2+)-activated Cl- currents in canine ventricular myocytes. Glibenclamide markedly inhibited CFTR Cl- currents in a voltage-independent manner at 22 degrees C, with estimated IC50 values of 12.5 and 11.0 mumol/L at +50 and -100 mV, respectively. The outwardly rectifying swelling-activated Cl- current in atrial cells was less sensitive to glibenclamide, and the block exhibited voltage dependence. At 22 degrees C, the estimated IC50 values were 193 and 470 mumol/L at +50 and -100 mV, respectively, and block was enhanced at 35 degrees C. Macroscopic Cl- currents activated by a rise in intracellular Ca2+, induced by either Ca(2+)-induced Ca2+ release or by external application of the Ca2+ ionophore A23187, were also markedly inhibited at 22 degrees C by glibenclamide in a voltage-independent manner. The estimated IC50 values were 61.5 and 69.9 mumol/L at +50 and -100 mV, respectively. These results suggest that glibenclamide, an inhibitor of cardiac CFTR Cl- channels, also inhibits swelling-activated and Ca(2+)-activated Cl- channels at higher concentrations. The results also suggest that studies attributing the beneficial or deleterious effects of sulfonylurea compounds in the heart solely to blockade of KATP channels should use submicromolar concentrations of these agents to minimize possible secondary interactions with cardiac Cl- channels. PMID- 9201034 TI - Alterations of Na+ currents in myocytes from epicardial border zone of the infarcted heart. A possible ionic mechanism for reduced excitability and postrepolarization refractoriness. AB - Previously, we have shown abnormalities in Vmax and in the recovery of Vmax in myocytes dispersed from the epicardial border zone (EBZ) of the 5-day infarcted canine heart (myocytes from the EBZ [IZs]). Thus, we sought to determine the characteristics of the whole-cell Na+ current (INa) in IsZs and compare them with the INa of cells from noninfarcted hearts (myocytes from noninfarcted epicardium [NZs]). INa was recorded using patch-clamp techniques under conditions that eliminated contaminating currents and controlled INa for measurement (19 degrees C, 5 mmol/L [Na+]zero). Peak INa density (at -25 mV) was significantly reduced in IZs (4.9 +/- 0.44 pA/pF, n = 36) versus NZs (12.8 +/- 0.55 pA/pF, n = 54; P < .001), yet the half-maximal activation voltage (V0.5), time course of decay, and time to peak INa were no different. However, in IZs, V0.5 of the availability curve (I/Imax curve) was shifted significantly in the hyperpolarizing direction ( 80.2 +/- 0.48 mV in NZs [n = 45] versus -83.9 +/- 0.59 mV in IZs [n = 27], P < .01). Inactivation of INa directly from a depolarized prepotential (-60 mV) was significantly accelerated in IZs versus NZs (fast and slow time constants [T1 and T2, respectively] were as follows: NZs [n = 28], T1 = 71.5 +/- 5.6 ms and T2 = 243.7 +/- 17.1 ms; IZs [n = 21], T1 = 36.3 +/- 2.4 ms and T2 = 153 +/- 11.3 ms; P < .001). Recovery of INa from inactivation was dependent on the holding potential (VH) in both cell types but was significantly slower in IZs. At (VH) = -90 mV, INa recovery had a lag in 18 (82%) of 22 IZs (with a 17.6 +/- 1.5-ms lag) versus 2 (9%) of 22 NZs (with 5.9- and 8.7-ms lags); at VH = -100 mV, T1 = 60.9 +/- 2.6 ms and T2 = 352.8 +/- 28.1 ms in NZs (n = 41) versus T1 = 76.3 +/- 4.8 ms and T2 = 464.4 +/- 47.2 ms in IZs (n = 26) (P < .002 and P < .03, respectively); at VH = -110 mV, T1 = 33.4 +/- 1.8 ms and T2 = 293.5 +/- 33.6 ms in NZs (n = 21) versus T1 = 44.3 +/- 2.9 ms and T2 = 388.4 +/- 38 ms in IZs (n = 18) (P < .002 and P < .07, respectively). In sum, INa is reduced, and its kinetics are altered in IZs. These changes may underlie the altered excitability and postrepolarization refractoriness of the ventricular fibers of the EBZ, thus contributing to reentrant arrhythmias in the infarcted heart. PMID- 9201035 TI - Developmental changes in transient outward current in mouse ventricle. AB - Developmental changes in the transient outward K+ current (Ito) in mouse ventricular myocytes were assessed by the whole-cell patch-clamp technique. The density of Ito in mouse ventricular myocytes was significantly increased from the day-1 neonate to the adult. At +50 mV, the density of Ito was 3 +/- 1 pA/pF in the day-1 neonate, 15 +/- 3 pA/pF in the day-14 neonate, and 19 +/- 4 pA/pF in the adult (P < .01). Unlike other species, the rate of Ito inactivation significantly slowed in mouse ventricular cells during development. Moreover, the time courses of inactivation and recovery from inactivation of Ito were well described by a monoexponential function in day-1 neonatal cells, whereas they were best fitted by a biexponential function in day-14 neonatal and adult cells. The characteristics of steady state inactivation were also significantly different in day-1 neonatal cells (half-inactivation potential [Vh] = -66 +/- 4 mV, slope factor [k] = 12 +/- 2 mV), in day-14 neonatal cells (Vh = -40 +/- 3 mV, k = 13 +/- 1 mV), and in adult cells (Vh = -34 +/- 4 mV, k = 6 +/- 1 mV). Microelectrode studies revealed that action potential duration progressively decreased in mouse ventricles during normal postnatal development. In addition, 4 aminopyridine (1 mmol/L) prolonged action potential duration more in adult than in neonatal mouse ventricles, suggesting that the developmental increase in the density of Ito contributes to the age-related shortening of action potential duration in mouse ventricles. In conclusion, Ito in adult mouse ventricular myocytes exhibits a higher density, slower inactivation kinetics, and a relatively more positive half-inactivation potential. All these characteristics result in Ito being a physiologically more important repolarizing K+ current in adult than in neonatal mouse hearts. PMID- 9201036 TI - Regional localization of ERG, the channel protein responsible for the rapid component of the delayed rectifier, K+ current in the ferret heart. AB - Repolarization of the cardiac action potential varies widely throughout the heart. This could be due to the differential distribution of ion channels responsible for repolarization, especially the K+ channels. We have therefore studied the cardiac localization of ERG, a channel protein known to play an important role in generation of the rapid component of the delayed rectifier K+ current (IKr), an important determinant of the repolarization waveform, Cryosections of the ferret atrium and ventricle were prepared to determine the localization of ERG by fluorescence in situ hybridization (FISH) and immunofluorescence. We found that in the ferret, ERG transcript and protein expression was most abundant in the epicardial cell layers throughout most of the ventricle, except at the base. In the atrium, we found that ERG is most abundant in the medial right atrium, especially in the trabeculae and the crista terminalis of the right atrial appendage. It also is present in areas within the sinoatrial node. In all regions studied, FISH and immunofluorescence showed concordant localization patterns. These data suggest that repolarization mediated by IKr is not uniform throughout the ferret heart and provide a molecular explanation for heterogeneity in action potential repolarization throughout the mammalian heart. PMID- 9201037 TI - Pulmonary Artery Catheter Consensus Conference. PMID- 9201038 TI - The flow-directed, pulmonary artery catheter and outcome in critically ill patients: have we heard the last word? PMID- 9201039 TI - Impaired oxygen extraction in sepsis: is supranormal oxygen delivery helpful? PMID- 9201040 TI - High-frequency oscillatory ventilation for adult respiratory distress syndrome: let's get it right this time! PMID- 9201041 TI - Recombinant tissue plasminogen activator restores perfusion in meningococcal purpura fulminans. PMID- 9201042 TI - Pulmonary Artery Catheter Consensus conference: consensus statement. PMID- 9201043 TI - Oxygen transport patterns in patients with sepsis syndrome or septic shock: influence of treatment and relationship to outcome. AB - OBJECTIVE: To investigate the relationship between oxygen transport patterns and outcome in patients with sepsis syndrome or septic shock managed according to two different treatment regimens. DESIGN: Retrospective study of a subgroup of patients with sepsis syndrome or septic shock taken from a randomized, prospective, controlled trial. SETTING: General intensive care units in a teaching and a district general hospital. PATIENTS: Seventy-eight patients classified according to predetermined criteria as having sepsis syndrome or septic shock were drawn retrospectively from a larger study group of 109 consecutive patients considered to be at risk for developing multiple organ failure. INTERVENTIONS: All patients received volume expansion to an optimal pulmonary artery occlusion pressure. If the therapeutic goals (cardiac index of > 4.5 L/min/m2, oxygen delivery [DO2] of > 600 mL/min/m2, and oxygen consumption [VO2] of > 170 mL/min/m2) were not achieved with fluids alone, patients were randomized to either a control group or a treatment group. In the treatment group, dobutamine (5 to 200 micrograms/kg/min) was administered to increase cardiac index and DO2 until all three goals were simultaneously achieved. In the control group, dobutamine was administered only if the cardiac index was < 2.8 L/min/m2. In both groups, norepinephrine was infused to maintain mean arterial pressure at 80 mm Hg. MEASUREMENTS AND MAIN RESULTS: Hemodynamic, oxygen transport, and lactate measurements were made at the time of admission to the study, at the time of optimal volume administration, at 1, 2, 4, 8, 12, 16, 20, and 24 hrs, then every 6 hrs for the next 24 hrs, and at least every 8 hrs thereafter. The time at which all therapeutic goals were first achieved simultaneously or the time of maximal DO2 was identified and termed "tmax." Survivors from both the control and treatment groups significantly (p < .001) increased cardiac index and DO2 in response to maximal resuscitation, and despite an associated decrease in oxygen extraction (p < .01), there was a significant (p < .01) increase in VO2. In nonsurvivors from both groups, despite significant increases in cardiac index (p < .05) and DO2 (p < .01) at tmax, oxygen extraction decreased (p < .01) and VO2 remained unchanged. DO2 and VO2 were significantly lower (p < .05) at tmax in nonsurvivors than in survivors from both groups. Persistently high lactate concentrations were characteristic of nonsurvivors. CONCLUSIONS: Survivors of sepsis syndrome or septic shock are characterized by an ability to increase both DO2 and VO2. In contrast, nonsurvivors typically have reduced cardiac reserve, they fail to increase VO2 following resuscitation, and when delivery is enhanced with aggressive inotropic support, oxygen extraction falls. These patterns of response were similar in both treatment and control groups, although the magnitude of the changes was exaggerated in the treatment group. These observations may help to explain the findings by some investigators that treatment aimed at achieving survivor values of cardiac index, DO2, and VO2 fails to improve outcome when instituted following admission to intensive care. PMID- 9201044 TI - High-frequency oscillatory ventilation for adult respiratory distress syndrome--a pilot study. AB - OBJECTIVE: To evaluate the safety and effectiveness of high-frequency oscillatory ventilation using a protocol designed to recruit and maintain optimal lung volume in patients with severe adult respiratory distress syndrome (ARDS). SETTING: Surgical and medical intensive care units in a tertiary care, military teaching hospital. DESIGN: A prospective, clinical study. PATIENTS: Seventeen patients, 17 yrs to 83 yrs of age, with severe ARDS (Lung Injury Score of 3.81 +/- 0.23) failing inverse ratio mechanical conventional ventilation (PaO2/FiO2 ratio of 68.6 +/- 21.6, peak inspiratory pressure of 54.3 +/- 12.7 cm H2O, positive end expiratory pressure of 18.2 +/- 6.9 cm H2O). INTERVENTIONS: High-frequency oscillatory ventilation was instituted after varying periods of conventional ventilation (5.12 +/- 4.3 days). We employed lung volume recruitment strategy that consisted of incremental increases in mean airway pressure to achieve a PaO2 of > or = 60 torr (> or = 8.0 kPa), with an FiO2 of < or = 0.6. MEASUREMENTS AND MAIN RESULTS: High-frequency oscillator ventilator settings (FiO2, mean airway pressure, pressure amplitude of oscillation [delta P] frequency) and hemodynamic parameters (cardiac output, oxygen delivery [DO2]), mean systemic and pulmonary arterial pressures, and the oxygenation index (oxygenation index = [FiO2 x mean airway pressure x 100]/PaO2) were monitored during the transition to high frequency oscillatory ventilation and throughout the course of the high-frequency protocol. Thirteen patients demonstrated improved gas exchange and an overall improvement in PaO2/FiO2 ratio (p < .02). Reductions in the oxygenation index (p < .01) and FiO2 (p < .02) at 12, 24, and 48 hrs after starting high-frequency oscillatory ventilation were observed. No significant compromise in cardiac output or DO2 was observed, despite a significant increase in mean airway pressure (31.2 +/- 10.3 to 34.0 +/- 6.7 cm H2O, p < .05) on high-frequency oscillatory ventilation. The overall survival rate at 30 days was 47%. A greater number of pretreatment days on conventional ventilation (p < .009) and an entry oxygenation index of > 47 (sensitivity 100%, specificity 100%) were associated with mortality. CONCLUSIONS: High-frequency oscillatory ventilation is both safe and effective in adult patients with severe ARDS failing conventional ventilation. A lung volume recruitment strategy during high-frequency oscillatory ventilation produced improved gas exchange without a compromise in DO2. These results are encouraging and support the need for a prospective, randomized trial of algorithm-controlled conventional ventilation vs. high-frequency oscillatory ventilation for adults with severe ARDS. PMID- 9201045 TI - Detection of Pneumocystis carinii in tracheal aspirates of intubated patients using calcofluor-white (Fungi-Fluor) and immunofluorescence antibody (Genetic Systems) stains. AB - OBJECTIVE: To compare the detection rate of Pneumocystis carinii in endotracheal aspirates with that rate in bronchoalveolar lavage fluid, using calcofluor-white (Fungi-Fluor) and immunofluorescence antibody (Genetic Systems) staining methods. DESIGN: Prospective, consecutive cases. SETTING: Medical intensive care unit at Ben Taub General Hospital. PATIENTS: Thirty-one intubated patients admitted with respiratory failure and suspected P. carinii pneumonia. INTERVENTIONS: An endotracheal aspirate specimen was obtained after maximally advancing a closed system suction catheter, instilling aliquot portions of saline, and suctioning the lavage fluid. This procedure was followed within 30 mins by fiberoptic bronchoscopy and bronchoalveolar lavage. MEASUREMENTS AND MAIN RESULTS: Endotracheal aspirate and bronchoalveolar lavage specimens from each patient were mixed with Saccomano's fixative, blended, and centrifuged. Using a modified method for P. carinii cysts, the sediment was stained with the test calcofluor white stain Solution A and the test antibody stain. The test antibody stain on the bronchoalveolar lavage specimens was positive for P. carinii for 13 patients and was used as the standard for comparison. In the endotracheal aspirate specimens, the test antibody stain detected 12 (92%) P. carinii-positive patients while the test calcofluor-white stain detected ten (77%) P. carinii-positive patients. CONCLUSIONS: We described a simple method for obtaining, processing, and staining endotracheal aspirate specimens for P. carinii. Obtaining an endotracheal aspirate specimen did not require specially trained personnel or a specialized and more expensive catheter, and was not associated with any complications. PMID- 9201046 TI - Increased plasma concentrations of adrenomedullin correlate with relaxation of vascular tone in patients with septic shock. AB - OBJECTIVE: To investigate plasma concentrations of adrenomedullin in patients with septic shock and the potential association of these concentrations with relaxation of vascular tone. DESIGN: Prospective, case series. SETTING: Department of Emergency and Critical Care Medicine, Nara Medical University. PATIENTS: Twelve patients who fulfilled the clinical criteria for severe sepsis or septic shock (as defined by the Members of the American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference Committee) and 13 healthy volunteers. INTERVENTIONS: Arterial blood samples were obtained via a 20-gauge cannula inserted into each patient's radial artery. MEASUREMENTS AND MAIN RESULTS: After extraction and purification, plasma adrenomedullin was measured by radioimmunoassay. Systemic vascular resistance index, pulmonary vascular resistance, cardiac index, and stroke volume index were determined with a thermodilution catheter. The mean plasma concentration of adrenomedullin was markedly higher in patients than in controls (226.1 +/- 66.4 [SEM] vs. 5.05 +/- 0.21 fmol/mL, p < .01). Moreover, these concentrations correlated significantly with cardiac index, stroke volume index, and heart rate values, and correlated significantly with decreases in diastolic blood pressure, systemic vascular resistance index, and pulmonary vascular resistance index values. CONCLUSIONS: Enhanced production of adrenomedullin in patients with septic shock may contribute to reduced vascular tone, hypotension, or both. More data are needed to clarify the role of adrenomedullin in the regulation of vascular tone in this patient population. PMID- 9201047 TI - Endothelin-1 and endothelin-4 stimulate monocyte production of cytokines. AB - OBJECTIVE: To determine the effect of endothelin-1 and endothelin-4 on human monocyte production of cytokines. DESIGN: Previous work from our laboratory has shown that endothelin-1 activates leukocytes. Endothelin-1 and endothelin-3 are principally produced by vascular endothelium. However, epidermal cells in gut mucosa, lung, and kidney produce endothelin-2 and endothelin-4, which differ by a single amino acid. While structurally similar to endothelin-1, endothelin-2 and endothelin-4 may affect gut smooth muscle and other tissues differently. The effect of endothelin-1 and endothelin-4 was examined on monocyte production of interleukins (IL) and neutrophil activation factors. SETTING: A clinically oriented basic science laboratory in a Veterans Administration Hospital and Medical Center. SUBJECTS: Healthy volunteer adult male/female medical students, researchers, and hospital workers. INTERVENTIONS: Human peripheral blood mononuclear cells were separated on density gradients and cultured in media, with or without the addition of bacterial endotoxin or varying molar concentrations of endothelin-1 and endothelin-4. Supernatants were harvested at 10 mins, and at 1, 6, 12, 24, and 48 hrs, and enzyme-linked immunosorbent assays were performed to determine the presence of tumor necrosis factor-alpha, IL-1 beta, IL-6, IL-8, and granulocyte-macrophage colony-stimulating factor. MEASUREMENTS AND MAIN RESULTS: Endothelin-1 and endothelin-4 were potent stimuli for monocyte production of tumor necrosis factor-alpha, IL-8, and granulocyte-macrophage colony-stimulating factor. They also caused IL-1 beta and IL-6 production. CONCLUSIONS: Endothelin-1 and endothelin-4 may activate leukocytes after shock or gut ischemia, resulting in further injury to reperfused tissues and distant injury to lungs and other organs. PMID- 9201048 TI - Small hemodynamic effect of typical rapid volume infusions in critically ill patients. AB - OBJECTIVES: To determine what volumes are commonly used for rapid volume infusions in critically ill patients admitted to the intensive care unit (ICU) for > 12 hrs; and to determine the effectiveness of a typical rapid volume infusion in producing hemodynamic change and increasing left ventricular end diastolic volume. DESIGN: A prospective survey of clinical practice (part 1) and a prospective clinical investigation (part 2). SETTING: Two hospital ICUs (11 and six beds) of which one is university affiliated. PATIENTS: Critically ill patients admitted to the ICU for > 12 hrs. INTERVENTIONS: Infusion of 500 mL of normal saline over 5 to 10 mins. MEASUREMENTS AND MAIN RESULTS: For 1 month, we recorded the volume and composition of all volume infusions given as a rapid bolus in patients admitted to the ICU for > 12 hrs. We then measured the effected the median rapid volume infusion in a subset of 13 patients by measuring hemodynamics (using arterial and pulmonary artery flotation catheters) and left ventricular end-diastolic area (using transgastric short-axis views from transesophageal echocardiograms). During 470 patient days, 159 rapid volume infusions were administered. The average rapid volume infusion administered was 390 +/- 160 mL (median 500; interquartile range 250 to 500). Crystalloid solutions were used for two thirds of the rapid volume infusions and colloid solutions were used for one third of the rapid volume infusions. The rapid volume infusion of 500 mL of saline did not significantly increase mean arterial pressure (78.0 +/- 11.9 to 79.3 +/- 14.6 mm Hg), cardiac index (4.3 +/- 1.7 to 4.6 +/- 1.8 L/min/m2), right atrial pressure (11.1 +/- 3.8 to 12.4 +/- 3.3 mm Hg), left ventricular end-diastolic area (8.6 +/- 1.7 to 9.1 +/- 1.8 cm2/m2), or left ventricular end-systolic area (3.5 +/- 1.5 to 3.6 +/- 1.5 cm2/m2). Pulmonary artery occlusion pressure increased slightly but significantly from 12.9 +/- 3.4 to 14.7 +/- 3.3 mm Hg (p < .05). CONCLUSIONS: After patients are admitted to the ICU for > 12 hrs, rapid volume infusions are common therapeutic interventions but the rapid volume infusions are typically small. The effect of a typical rapid volume infusion on hemodynamics and left ventricular areas in these patients is surprisingly small. PMID- 9201049 TI - Delayed hyperemia causing intracranial hypertension after cardiopulmonary resuscitation. AB - OBJECTIVE: To clarify whether early or delayed failure of cerebral perfusion after cardiopulmonary resuscitation (CPR) occurs in humans and contributes to secondary brain damage. DESIGN: Prospective, repeated-measures study. SETTING: Intensive care unit of Hiroshima University School of Medicine. PATIENTS: Eight comatose patients who had undergone successful resuscitation from cardiac arrest. INTERVENTIONS: All patients underwent transcranial Doppler sonography examination. The intracranial cerebral pressure (ICP) and jugular venous oxygen saturation (SO2) also were continuously monitored in five patients and three patients, respectively. MEASUREMENTS AND MAIN RESULTS: In each patient, we measured the mean flow velocity of the middle cerebral artery transcranially and the mean flow velocity of the internal carotid artery, high in the neck, using transcranial Doppler sonography. The pulsatility index for each measurement was also calculated. The first examinations were performed within 4 to 12 hrs of CPR, and repeat examinations were performed approximately every 12 hrs. The initial mean flow velocities of the middle cerebral artery and the initial mean flow velocities of the internal carotid artery were relatively low, with relatively high pulsatility indices. The mean flow velocities of the middle cerebral artery began to increase at 12 to 24 hrs after CPR and peaked 24 to 120 hrs after CPR. A simultaneous increase in mean flow velocities of the internal carotid artery was observed during this period. The pulsatility index in both arteries dropped significantly during peak mean flow velocity of the middle cerebral artery. In six of seven patients with an abnormal increase (> 100 cm/ sec) in peak mean flow velocity of the middle cerebral artery, the ratio of mean flow velocity of the middle cerebral artery to mean flow velocity of the internal carotid artery was < 3. This value tended to be lower in patients with poor outcomes. An increased mean flow velocity of the middle cerebral artery, with a ratio of < 3 for mean flow velocity of the middle cerebral artery to mean flow velocity of the internal carotid artery, was defined as hyperemia. Although the mean flow velocity of the internal carotid artery was not measured, another patient with an abnormal increase in mean flow velocity of the middle cerebral artery revealed a high jugular venous SO2 value of 83.5%, also representing hyperemia. All ICP values were within the normal range 4 to 12 hrs after CPR and tended to increase before peak mean flow velocity of the middle cerebral artery. The two patients with the lowest ratios of mean flow velocity of the middle cerebral artery to mean flow velocity of the internal carotid artery showed significant increases in ICP after the peak mean flow velocity of the middle cerebral artery. These two patients subsequently developed brain death. CONCLUSIONS: Delayed hyperemia occurs in humans after resuscitation from cardiac arrest. Our data suggest that this delayed hyperemia can lead to intracranial hypertension and occasionally acute brain swelling, contributing to a poor outcome. A high mean flow velocity of the middle cerebral artery with a low ratio of mean flow velocity of the middle cerebral artery to mean flow velocity of the internal carotid artery may be predictive of critical hyperemia. As an indirect method of measuring cerebral blood flow transcranial Doppler sonography can be used to adjust treatment for failure of cerebral perfusion after resuscitation. PMID- 9201051 TI - Clinical and economic outcome of patients undergoing tracheostomy for prolonged mechanical ventilation in New York state during 1993: analysis of 6,353 cases under diagnosis-related group 483. AB - OBJECTIVE: To examine and describe the relation between age and disposition in patients undergoing tracheostomy. DESIGN: Retrospective analysis of a statewide database. SETTING: All acute care hospitals in New York state. PATIENTS: All patients (n = 6,353) > or = 18 yrs of age who were discharged from the hospital during 1993 with a final diagnosis-related groups code of 483. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The final disposition, according to six disposition codes (other acute care facility, residential healthcare facility, other healthcare facility, home, home healthcare services, and death) was examined for the entire population. Cost per case was assumed to equal the average statewide Medicaid rate. An inverse relation between survival rate and age was observed, which resulted in an age-related increased cost per survivor. Also, survivors in older age groups had an increased rate of discharge to residential healthcare facilities. There was a negative, albeit less marked, effect of older age on the rates of survivors discharged to home and to other healthcare facilities. CONCLUSIONS: Care of patients who undergo tracheostomy for prolonged mechanical ventilation is expensive. The older the patient, the less satisfactory the outcome from an economic, clinical, and possibly social perspective. PMID- 9201050 TI - Prolonged extracorporeal life support (ECLS) for varicella pneumonia. AB - OBJECTIVE: To review the institutional experience of a national tertiary referral center for extracorporeal life support (ECLS) in severe varicella pneumonia. DATA SOURCES: Hospital records and ECLS flow sheets. STUDY SELECTION: All pediatric (nonneonatal) and adult patients who were treated for varicella pneumonia with ECLS at the University of Michigan Medical Center between 1986 and 1995. DATA EXTRACTION: Diagnosis of varicella pneumonia was made by history of recent exposure to chickenpox, progressive dyspnea, fever, a characteristic diffuse, vesicular rash, and a supporting chest roentgenogram. Indications for ECLS included a shunt fraction of > 30% or PaO2/FlO2 ratio of < 80 despite maximal conventional therapy, which included aggressive diuresis, blood transfusions to optimize oxygen-carrying capacity, pressure-controlled/inverse-ratio ventilation, and intermittent prone positioning. DATA SYNTHESIS: Between 1986 and 1995, 191 patients were referred for ECLS. Among these patients, there were 51 (27%) cases of viral pneumonia, of which nine cases were due to acute varicella-zoster infection. Intravenous acyclovir was administered to eight of the nine patients. Of the nine patients, two patients improved using conventional ventilator management, and seven patients underwent ECLS. Overall survival on ECLS was 71% (5/7). The mean (+/-SD) alveolar-arterial oxygen gradient and PaO2/FlO2 ratio were 533 +/- 101 torr (71.3 +/- 13.5 kPa) and 67 +/- 24, respectively. The median duration of mechanical ventilation before ECLS and the subsequent duration of ECLS were 4 and 12.8 days, respectively. One of the deaths was from progressive right heart failure secondary to pulmonary hypertension and the other death was from overwhelming Pseudomonas sepsis. CONCLUSIONS: Early recognition of imminent pulmonary failure and rapid institution of ECLS are critical in the successful management of severe, life-threatening varicella pneumonia. PMID- 9201052 TI - Utilization and diagnostic yield of blood cultures in a surgical intensive care unit. AB - OBJECTIVE: To evaluate the diagnostic yield of blood cultures obtained in a surgical intensive care unit (ICU) and to assess factors potentially influencing yield. DESIGN: Retrospective, descriptive study. SETTING: Surgical ICU in a university hospital. SUBJECTS: All patients who had a blood culture obtained during their admission to the trauma/neurosurgical ICU of Presbyterian University Hospital from January 1, 1993 to December 31, 1993. MEASUREMENTS AND MAIN RESULTS: Blood culture isolates were categorized as pathogens or contaminants and overall diagnostic yield was determined. Blood cultures were positive for pathogens in 4.6% of all culture episodes, while contaminants were isolated in 5.5% of all culture episodes. A total of 23 true bacteremias were identified in 21 patients, for an overall rate of bacteremia of 3.6 per 100 admissions (5.9 per 1,000 patient days). Concurrent antibiotics were being used at the time of blood culture in 65.3% of all culture episodes. The yield for pathogens was significantly lower (2.2%) when cultures were obtained on antibiotics compared with culture episodes obtained off antibiotics (6.4%) (p < .05). Single-set blood culture episodes were obtained in approximately 32% of all culturing episodes with the overall yield for pathogens of these culturing episodes lower (2.9%) than that of multiple-set culture episodes (5.3%) (p = NS). CONCLUSIONS: Blood culture yield in this surgical ICU was relatively low in comparison with other published studies. The data further suggest that concurrent use of systemic antibiotics and inappropriate or excessive culturing may negatively influence blood culture yield. PMID- 9201053 TI - Variation of inhaled nitric oxide concentration with the use of a continuous flow ventilator. AB - OBJECTIVE: To investigate the homogeneity of nitric oxide concentrations at different ventilator settings in a delivery system using a continuous flow ventilator. DESIGN: A prospective bench study using a nitric oxide delivery system, mixing a nitrogen/nitric oxide gas mixture in the ventilator circuit with two types of continuous flow ventilators (Babylog 8000, Draeger, Luebeck, Germany; Infant Star, Infrasonics, San Diego, CA). SETTING: A biomedical laboratory. INTERVENTIONS: A nitrogen/nitric oxide gas mixture was injected at three different sites in the ventilator circuit (just before and just behind the humidifier, and 20 cm before the Y-connector). Ventilator flow (12, 15, and 20 L/min) and rates (30 to 110 breaths/min with increments of 10 breaths/min) were changed as well as the compliance of the test lung (0.36, 0.5, and 1.0 mL/cm H2O). Carbon dioxide, instead of nitrogen/nitric oxide, was injected at the same points in the circuit. MEASUREMENTS AND MAIN RESULTS: The mean nitric oxide concentration increased significantly (p < .001) with increasing ventilator rates (although the flow ratio of the ventilator gas and the nitrogen/nitric oxide gas mixture was kept constant) when the nitrogen/nitric oxide injection site was near to the Y-connector of the ventilator circuit with both ventilators. The mean nitric oxide concentration did not change significantly when the nitrogen/nitric oxide gas mixture was mixed to the ventilator gas at the inlet of the humidifier, using the Babylog 8000 ventilator. Analysis of ventilator circuit flow patterns showed fluctuations during the respiratory cycle. The magnitude of the flow changes was different at the three injection sites in the ventilator circuit. Real-time measurements of the CO2 concentration showed fluctuations during the distinct respiratory phases that differed at the separate injection sites. Mean CO2 concentrations showed a similar pattern as compared with the mean nitric oxide concentration data at the same settings. CONCLUSIONS: Mixing a nitrogen/nitric oxide gas mixture 20 cm before the Y-connector results in an increase of the mean nitric oxide concentration with increasing ventilator rates. This phenomenon does not occur with the nitrogen/nitric oxide gas mixture mixed at the inlet of the humidifier, using a ventilator with a throughout constant flow at the inspiratory outlet of the ventilator. The fluctuations of the main ventilator circuit flow result in changing ratios of nitrogen/nitric oxide gas mixture and the ventilator gas flow. We speculate this changing flow ratio produces the increase in mean nitric oxide concentration within the ventilatory circuit. To ensure a constant concentration of nitric oxide by blending a nitrogen/nitric oxide gas mixture in the ventilator circuit of a continuous flow ventilator, the site of injection of the nitrogen/nitric oxide gas mixture should be at the point where ventilator circuit flow fluctuations are minimal. PMID- 9201054 TI - Effects of intra-aortic balloon occlusion on hemodynamics during, and survival after cardiopulmonary resuscitation in dogs. AB - OBJECTIVE: To evaluate the effect of balloon occlusion of the proximal descending aorta during cardiopulmonary resuscitation (CPR) on hemodynamics, restoration of spontaneous circulation, and 24-hr survival. DESIGN: Prospective, randomized, controlled trial. SETTING: Experimental laboratory in a university hospital. SUBJECTS: Eighteen anesthetized dogs. INTERVENTIONS; Catheters were placed for hemodynamic and blood gas monitoring. An aortic balloon catheter was placed with its tip just distal to the left subclavian artery. After 10 mins of ventricular fibrillation without CPR, 3 mins of Basic Life Support (chest compressions and ventilation with 100% oxygen) was followed by up to 30 mins of Advanced Cardiac Life Support with canine drug dosages. In the treatment group (n = 8), the intra aortic balloon was inflated when Advanced Cardiac Life Support started and not deflated until shortly after restoration of spontaneous circulation. The control animals (n = 10) were treated with an identical resuscitation but without intra aortic balloon occlusion. MEASUREMENTS AND MAIN RESULTS: In the treatment group, coronary perfusion pressure was greater during Advanced Cardiac Life Support (p = .026). Restoration of spontaneous circulation was more frequent (7/8 dogs) as compared with the control group (3/10 dogs) (p = .025). There was a trend toward greater 24-hr survival in the treatment group (5/8 dogs) than in the control group (3/10 dogs). CONCLUSIONS: Balloon occlusion of the proximal descending aorta during experimental CPR improves restoration of spontaneous circulation. Further laboratory and human studies are needed to determine the clinical efficacy of this technique. PMID- 9201055 TI - Oxalated pyridoxalated hemoglobin polyoxyethylene conjugate normalizes the hyperdynamic circulation in septic sheep. AB - OBJECTIVE: Excessive production of nitric oxide significantly contributes to the hyperdynamic state associated with sepsis. The ability of hemoglobin to scavenge nitric oxide may therefore be beneficial in the treatment of sepsis. In this study, we determined the effects of different doses of the modified human pyridoxalated hemoglobin polyoxyethylene conjugate in an ovine model of hyperdynamic sepsis. DESIGN: Prospective, experimental study. SETTING: Large animal research laboratory at a university medical center. INTERVENTIONS: Sheep (n = 23) were surgically prepared for chronic study. After a 5-day recovery period, all animals received a continuous infusion of live Pseudomonas aeruginosa (2.5 x 10(6) colony-forming units/min) for the next 48 hrs. After 24 hrs of sepsis, the animals were divided into four groups: a) six sheep were used as controls and received a bolus of 200-mL vehicle; b) three sheep received a bolus of 50 mg/kg hemoglobin; c) six sheep received 100 mg/kg of hemoglobin; d) six sheep received 200 mg/kg of hemoglobin. MEASUREMENTS AND MAIN RESULTS: All animals that survived the first 24 hrs of sepsis (n = 21) developed a hyperdynamic circulation. All three doses of hemoglobin reversed this hyperdynamic state by increasing mean arterial pressure and systemic vascular resistance while decreasing cardiac index. Pulmonary arterial pressure increased after hemoglobin infusion. Increased pulmonary arterial pressure did not affect arterial oxygen saturation nor result in the development of pulmonary edema. Infusion of hemoglobin also caused a 30-fold increase in endothelin-1 plasma concentrations and significantly decreased nitrate and nitrite plasma concentrations. CONCLUSIONS: The infusion of low doses of pyridoxalated hemoglobin polyoxyethylene conjugate in septic sheep reverses the hyperdynamic circulatory state. An increase in pulmonary arterial pressure was the only observed hemodynamic side effect; changes in the structure or function of other organ systems, or their biochemical correlates were not investigated in this study. In addition to a possible nitric oxide scavenging effect, pyridoxalated hemoglobin polyoxyethylene may affect the nitric oxide synthase and endothelin systems. PMID- 9201056 TI - Effects of carboxy-PTIO on systemic hemodynamics, liver energetics, and concentration of liver metabolites during endotoxic shock in rabbits: a 31P and 1H magnetic resonance spectroscopic study. AB - OBJECTIVE: To investigate the effects of 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO), a nitric oxide scavenger, on the lipopolysaccharide-induced hypotension, hepatocellular dysfunction, and liver damage in endotoxic rabbits. DESIGN: Experimental, comparative study. SETTING: Laboratory of a university hospital. SUBJECTS: Eighteen Japanese white rabbits (3.0 to 3.2 kg body weight) anesthetized with ketamine-xylazine were studied. INTERVENTIONS: We randomly divided the rabbits into three groups: saline controls (group 1, n = 5); animals receiving lipopolysaccharide (400 micrograms/kg) alone (group 2, n = 8); and animals receiving lipopolysaccharide plus carboxy-PTIO at a rate of 0.17 mg/kg/min for 3 hrs (group 3, n = 5). Blood gases and mean arterial pressure (MAP) were monitored. In vivo phosphorus-31 magnetic resonance spectra were continuously obtained every 30 mins. In addition, the livers were sampled and underwent fractionation at 7 hrs after lipopolysaccharide administration. The hydrophilic and hydrophobic extracts from the livers were analyzed by in vitro hydrogen-1 and phosphorus-31 magnetic resonance spectroscopy. MEASUREMENTS AND MAIN RESULTS: After the administration of lipopolysaccharide, the first phase of decrease in MAP within 30 mins was followed by partial recovery within the next 30 mins. In group 2, MAP started to decrease progressively within 180 mins after lipopolysaccharide administration (second phase) and decreased by 33% from the baseline value to 49 +/- 9 mm Hg at 420 mins. In contrast, the infusion of carboxy-PTIO significantly attenuated the second decrease in MAP (68 +/- 10 mm Hg, at 420 mins). In group 2, a slow and progressive decrease in adenosine triphosphate (ATP) and increase in inorganic phosphate concentrations occurred from 120 mins after lipopolysaccharide administration, and continued throughout the observation period. These changes were accompanied by a progressive decrease in intracellular pH. On the other hand, in group 3, there were no significant changes in ATP and inorganic phosphate concentrations compared with the controls from 120 to 360 mins after lipopolysaccharide administration. Moreover, restorations of both arterial and hepatocellular acidosis were observed in group 3. The differences of the degree of liver damage--as determined by the total amount of phospholipid, free fatty acids concentration, and membrane fluidity--were not significant among the three groups. Three of eight rabbits in group 2 died within 7 hrs, but no animal in the other two groups died during the study. CONCLUSIONS: The results of this study indicate that the infusion of carboxy-PTIO: a) prevented the delayed hypotension associated with endotoxic shock in rabbits; b) returned the hepatocellular ATP concentrations nearly to the level of the controls and alleviated hepatocellular acidosis; c) normalized various hydrophilic metabolites, such as lactate and alanine in the liver; and d) did not exacerbate liver injury after the administration of lipopolysaccharide. These findings indicate that carboxy-PTIO, a nitric oxide scavenger, may have a positive vasopressor effect during hypodynamic septic shock without exacerbating liver injury. PMID- 9201057 TI - Monoclonal antibody to endotoxin attenuates hemorrhage-induced lung injury and mortality in rats. AB - OBJECTIVES: To determine the possible role of enteric bacteria-derived endotoxin in the pathogenesis of the lung injury and mortality in rats following hemorrhagic shock and resuscitation. DESIGN: Prospective, randomized study. SETTING: Animal laboratory of an institute for research traumatology. SUBJECTS: Male Sprague-Dawley rats, weighing 450 to 480 g. INTERVENTIONS: Anesthetized rats were subjected to a prolonged hemorrhagic shock (mean arterial pressure of 30 to 35 mm Hg for 180 mins) followed by resuscitation. A murine monoclonal antibody to lipopolysaccharide from Escherichia coli and Salmonella, WN1 222-5, was administered at a total dose of 5 mg/kg i.v., starting at the onset of shock (WN1 group). The control group was treated similarly to the WN1 group but received saline at the same volume as WN1 222-5. MEASUREMENTS AND MAIN RESULTS: The 48-hr mortality rate was significantly reduced by WN1 222-5 treatment (28.6% in the treatment group vs. 78.6% in the control group; p = .0169). The characteristic lung injury in this model was significantly reduced in the WN1 group, as assessed by microscopic histopathologic examination increase in lung wet weight (7.60 +/- 0.47 g/kg in the control group vs. 5.14 +/- 0.31 g/kg in the WN1 group; p = .0002), and pulmonary neutrophilic infiltration (myeloperoxidase activity: 1835 +/- 567 mU/g wet weight in the control group vs. 891 +/- 212 mU/g wet weight in the WN1 group). CONCLUSIONS: These data suggest that a) endotoxin derived from enteric bacteria might play an important role in the pathogenesis of lung injury; and b) antiendotoxin agents, such as WN1 222-5, appear to protect against endogenous bacterial endotoxin-related disorders in severe hemorrhagic shock in rats. PMID- 9201058 TI - Comparative analysis of brain protection by N-methyl-D-aspartate receptor antagonists after transient focal ischemia in cats. AB - OBJECTIVE: We tested the hypothesis that the administration of the competitive N methyl-D-aspartate (NMDA) receptor antagonist 2R,4R,5S-(2-amino-4,5-(1,2 cyclohexyl)-7-phosphonoheptanoic acid) (NPC 17742) or cis-4-(phosphonomethyl) piperidine-2-carboxylic acid (CGS 19755) or the noncompetitive NMDA receptor antagonist dizocilpine (MK-801), at the appropriate doses, would all have efficacy in decreasing early postischemic brain injury in a feline model of transient focal ischemia. DESIGN: Prospective, randomized, controlled animal trial. SETTING: University research laboratory. SUBJECTS: Forty mixed-breed cats. INTERVENTIONS: Halothane-anesthetized cats underwent 90 mins of left middle cerebral artery occlusion plus 4 hrs of reperfusion. At 75 mins of ischemia, control cats received intravenous saline (n = 10). Experimental cats (n = 10 in each group) were treated with NPC 17742 (5 mg/kg bolus and 2.5 mg/kg/hr throughout reperfusion), MK-801 (5 mg/kg intravenous bolus), or CGS 19755 (40 mg/kg intravenous bolus) in a randomized fashion. MEASUREMENTS AND MAIN RESULTS: Microsphere-determined blood flow to the ipsilateral inferior temporal cortex and caudate nucleus decreased to the same extent during ischemia, and recovered to the same extent during early reperfusion, in the four groups. Triphenyltetrazolium-determined injury volume of the ipsilateral caudate nucleus in cats treated with NPC 17742 (105 +/- 25 [SEM] mm3), MK-801 (97 +/- 22 mm3), and CGS 19755 (97 +/- 13 mm3) was less than in control cats (198 +/- 21 mm3). Hemisphere injury volumes with NPC 17742 (1209 +/- 405 mm3) and MK-801 (1338 +/- 395 mm3) were less than that value in controls (2193 +/- 372 mm3), whereas injury volume with CGS 19755 (1553 +/- 519 mm3) treatment did not attain significance (p < .09). CONCLUSIONS: NMDA receptor activation during reperfusion may contribute to the progression of injury in ischemic border regions after 90 mins of transient focal ischemia in the cat. At the doses chosen, there appear to be no major differences in therapeutic efficacy for competitive and noncompetitive NMDA receptor antagonists. PMID- 9201059 TI - Xanthine oxidase mediates myocardial injury after hepatoenteric ischemia reperfusion. AB - OBJECTIVES: To determine if myocardial injury results from hepatoenteric ischemia reperfusion. We also proposed to determine if this remote heart injury is mediated by a xanthine oxidase-dependent mechanism. DESIGN: Randomized, controlled animal study. SETTING: University-based animal research facility. SUBJECTS: Thirty-six New Zealand white male rabbits, weighing 1.8 to 3 kg. INTERVENTIONS: Anesthetized rabbits were randomly assigned to one of four groups (n = 9 per group): a) a sham-operated group; b) a sham-operated group pretreated with sodium tungstate (xanthine oxidase inactivator); c) an aorta occlusion group; and d) an aorta occlusion group pretreated with sodium tungstate. Descending thoracic aorta occlusion was maintained for 40 mins with a 4-Fr Fogarty embolectomy catheter, followed by 2 hrs of reperfusion. MEASUREMENTS AND MAIN RESULTS: Myocardial injury, manifested by increased circulating creatine kinase-MB fraction activity, was significantly associated with aortic occlusion and reperfusion (p < .05). Sodium tungstate pretreatment significantly (p < .05) reduced circulating and myocardial xanthine oxidase activity. Xanthine oxidase inactivation by sodium tungstate significantly decreased circulating creatine kinase-MB fraction activity after hepatoenteric ischemia-reperfusion (p < .05). Finally, circulating creatine kinase-MB fraction activity was significantly associated with circulating xanthine oxidase activity (r2 = .85; p < .001). CONCLUSIONS: We conclude that remote myocardial injury is caused by hepatoenteric ischemia-reperfusion. The pathoetiology of this myocardial injury involves a xanthine oxidase-dependent mechanism. PMID- 9201060 TI - Effect of NG-nitro-L-arginine-methyl-ester on cardiopulmonary function and biosynthesis of cyclooxygenase products during porcine endotoxemia. AB - OBJECTIVE: To determine if inhibition of nitric oxide synthase with NG-nitro-L arginine-methyl-ester (L-NAME) potentiates endotoxin-induced cardiopulmonary dysfunction and release of cyclooxygenase products in a porcine model of endotoxemia. DESIGN: Prospective, multiple group, controlled experimental study. SETTING: Physiologic research laboratory at a veterinary medicine college. SUBJECTS: Fifty-seven domestic pigs (mean 28.7 +/- 0.8 [SEM] kg). INTERVENTIONS: Pentobarbital-anesthetized pigs were intubated and mechanically ventilated to normocapnia with room air. A ther-modilution cardiac output catheter was advanced into the pulmonary artery. Additional catheters were inserted into the jugular and femoral veins and femoral artery. The pigs received the following infusions: saline (control, n = 5); L-NAME (0.1, 0.5, 2.2, or 5.5 mg/ kg/hr, from -0.5 to 2 hrs, n = 16); Escherichia coli endotoxin (5 micrograms/ kg from 0 to 1 hr followed by 2 micrograms/kg from 1 to 2 hrs, i.v., n = 14); L-NAME plus endotoxin (n = 9); indomethacin plus endotoxin (n = 6); or L-NAME indomethacin plus endotoxin (n = 7). MEASUREMENTS AND MAIN RESULTS: L-NAME significantly (p < .05) worsened endotoxin-induced hypoxemia and enhanced the increases in pulmonary vascular resistance index and systemic vascular resistance index at 30 to 60 mins. Endotoxin increased (p < .05) plasma concentrations of thromboxane B2 by seven- to eight-fold at 30 to 120 mins and 6-keto-prostaglandin F1 alpha by 16- to 24-fold at 60 to 120 mins. L-NAME enhanced (additive effect) endotoxin-induced increases in plasma concentrations of thromboxane B2 (60 mins) and significantly (p < .05) potentiated the increases in 6-keto-prostaglandin F1 alpha (120 mins). At 120 mins of endotoxemia, indomethacin (cyclooxygenase inhibitor) plus L-NAME markedly increased (p < .05, synergistic effect) systemic vascular resistance index compared with endotoxemic pigs pretreated with either L-NAME or indomethacin. CONCLUSIONS: During endotoxemia, inhibition of nitric oxide synthase with L-NAME may be deleterious to cardiopulmonary function, as evidence by potentiation of endotoxin-induced systemic and pulmonary vasoconstriction, impairment of gas exchange, and enhanced biosynthesis of cyclooxygenase products. Moreover, during endotoxemia, the concomitant inhibition of two important vasodilators (i.e., nitric oxide and prostacyclin) is associated with a potentiated (p < .05) increase in systemic vascular resistance index. PMID- 9201061 TI - Effects of varying levels of positive end-expiratory pressure on intracranial pressure and cerebral perfusion pressure. AB - OBJECTIVE: To determine the influence of positive end-expiratory pressure (PEEP) on intracranial pressure and cerebral perfusion pressure. DESIGN: Neurosurgical intensive care patients requiring intracranial pressure monitoring and mechanical ventilation were studied in a randomized, controlled study. SETTING: Tertiary care, neurosurgical intensive care unit. PATIENTS: Eighteen patients were enrolled in the study. Patients had posttraumatic head injuries (n = 9), subarachnoid hemorrhage (n = 7), obstructive hydrocephalus (n = 1), and intracerebral hemorrhage of unknown cause (n = 1). INTERVENTIONS: Patients had PEEP levels of 5, 10, and 15 cm H2O applied to their lungs. MEASUREMENTS AND MAIN RESULTS: Changes in intracranial pressure, mean arterial pressure, and cerebral perfusion pressure were measured. The results were analyzed separately for patients with normal and increased intracranial pressure (> 15 mm Hg). PEEP at 5 cm H2O had no effect on intracranial pressure in the group with normal intracranial pressure. However, PEEP at 10 and 15 cm H2O produced a significant (p < .05) increase in intracranial pressure (1.9 and 1.5 mm Hg, respectively). In the group with increased intracranial pressure, no significant change in intracranial pressure occurred at any of the PEEP levels used. In both groups, cerebral perfusion pressure was unchanged throughout. CONCLUSIONS: In patients with normal intracranial pressure, PEEP at 5 cm H2O did not significantly alter intracranial pressure. The clinical relevance of the intracranial pressure increase at PEEP levels of 10 and 15 cm H2O is questionable because cerebral perfusion pressure did not change and remained > 60 mm Hg. In patients with increased intracranial pressure, higher levels of PEEP did not significantly change intracranial pressure or cerebral perfusion pressure. PMID- 9201062 TI - Endogenous nitric oxide production and atrial natriuretic peptide biological activity in infants undergoing cardiac operations. AB - OBJECTIVES: To examine whether preoperative heart failure and cardiac surgery influence nitric oxide production and atrial natriuretic peptide (ANP) biological activity in infants and whether nitric oxide and ANP participate in the control of postoperative pulmonary vascular tone. DESIGN: Prospective, clinical study. SETTING: Tertiary pediatric cardiac intensive care unit in a referral cardiosurgical center. PATIENTS: Nineteen infants (median age 4 months) undergoing cardiac surgery: 13 infants with ventricular or atrioventricular septal defect associated with heart failure and pulmonary hypertension (group 1); and six infants with tetralogy of Fallot, without heart failure (group 2). INTERVENTIONS: Blood samples obtained from indwelling catheters or bypass circuit outlets. MEASUREMENTS AND MAIN RESULTS: Nitrite and nitrate blood concentrations (as a marker for nitric oxide synthesis) and the molar ratio of cyclic guanosine 3',5'-monophosphate (cGMP) to ANP (as a marker for ANP biological activity) were determined before, during, and up to 24 hrs after cardiopulmonary bypass (CPB). In group 1 patients, these biological parameters were related to postoperative pulmonary arterial pressure. Preoperative nitrite and nitrate concentrations were higher in group 1 patients than in group 2 patients (p < .02), and this difference persisted during CPB. Nitrite and nitrate concentrations 24 hrs postoperatively were lower than preoperative values in group 1 patients (p < .05) and were unchanged in group 2 patients. An inverse correlation was observed postoperatively between nitrite and nitrate concentrations and systolic pulmonary arterial pressure (r2 = 0.4, p < .05). Group 1 patients had a lower preoperative cGMP/ANP ratio than group 2 patients (p < .05), despite higher ANP levels (p < .005). The cGMP/ANP ratio decreased during CPB in both groups (p < .0001), and in group 2 patients, cGMP and ANP values remained below preoperative values < or = 24 hrs postoperatively. A correlation was observed between ANP levels and systolic pulmonary arterial pressure 2 and 4 hrs postoperatively (r2 = .4, p < .05, respectively), but no correlation was observed between ANP biological activity and postoperative pulmonary arterial pressure. CONCLUSIONS: Infants with heart failure and pulmonary hypertension have increased nitric oxide synthesis and decreased ANP biological activity; both phenomena may be involved in the pathophysiology of this clinical condition. CPB has no detectable effect on nitric oxide production but does decrease ANP biological activity. In patients with preoperative heart failure and pulmonary hypertension, endogenous nitric oxide appears to play a role in the control of postoperative pulmonary vascular tone. PMID- 9201064 TI - Recombinant tissue plasminogen activator restores perfusion in meningococcal purpura fulminans. AB - OBJECTIVE: To investigate whether an infusion of recombinant tissue plasminogen activator would dissolve microvascular thromboses and improve organ perfusion in a patient with fulminant meningococcemia. DESIGN: Descriptive case report. SETTING: Fifteen-bed pediatric intensive care unit (ICU) in a university hospital. PATIENT: A 4-month-old male with fulminant meningococcemia, refractory shock, and multiple organ failure. INTERVENTIONS: In addition to standard aggressive ICU care, the patient received a recombinant tissue plasminogen activator infusion at a total dose of 1.25 mg/kg over 4 hrs. MEASUREMENTS AND MAIN RESULTS: Heart rate, arterial blood pressure, urine output, and base deficit (as a reflection of severity of metabolic acidosis) were recorded immediately before the recombinant tissue plasminogen activator infusion and 4 hrs later, after completion of the recombinant tissue plasminogen activator infusion. The amount of exogenous vasopressor and inotropic support required to maintain the patient's hemodynamic status before and after recombinant tissue plasminogen activator infusion were also compared. Subjective observations regarding the patient's peripheral perfusion status were also noted. The patient showed a dramatic improvement in hemodynamics, urine output, and metabolic acidosis, as well as a perceived increase in skin perfusion after recombinant tissue plasminogen activator infusion. CONCLUSIONS: In this patient, recombinant tissue plasminogen activator infusion resulted in improved organ perfusion and cardiac performance. Selective use of recombinant tissue plasminogen activator in the treatment of fulminant meningococcemia merits further investigation. PMID- 9201063 TI - Measurements of total plasma nitrite and nitrate in pediatric patients with the systemic inflammatory response syndrome. AB - OBJECTIVES: The systemic inflammatory response syndrome (SIRS) is typified by the presence of fever, hemodynamic changes, and end organ dysfunction. Endothelial cell activation leads to overproduction of nitric oxide, which results in sustained vasodilation and hypotension. This study was undertaken to determine the sensitivity, specificity, and positive and negative predictive values of plasma nitrite/nitrate measurements in identifying patients with clinical characteristics of SIRS, as defined by criteria based on physician diagnosis. DESIGN: Prospective cohort study with consecutive sampling of patients. SETTING: Tertiary, multidisciplinary, pediatric intensive care unit (ICU) at Children's Hospital of Wisconsin. PATIENTS: Patients were divided into five groups. There were 16 pediatric controls undergoing elective surgery and 177 pediatric ICU patients without and 46 pediatric ICU patients with physician-diagnosed sepsis, septic shock, SIRS, or sepsis syndrome documented in the medical record (all considered physician-diagnosed sepsis). The 223 pediatric ICU patients included 195 pediatric ICU patients not meeting and 28 pediatric ICU patients meeting predetermined physiologic criteria for SIRS (considered criteria-based sepsis). INTERVENTIONS: Blood samples were obtained for quantitative nitrite/nitrate analysis at the time of admission to the pediatric ICU and daily until discharge. MEASUREMENTS AND MAIN RESULTS: Mean plasma nitrite/nitrate concentrations in the controls were 34.5 +/- 12 microM (95th percentile 54 microM). In pediatric ICU patients without and with physician-diagnosed sepsis, mean plasma nitrite/nitrate concentrations were 39 +/- 24 microM (p > .05 compared with controls) and 127 +/- 91 microM (p < .0001 compared with both controls and patients without physician diagnosed sepsis), respectively. In pediatric ICU patients without and with criteria-based sepsis, the mean total plasma nitrite/nitrate concentrations were 56 +/- 59 microM (p = .008 compared with controls) and 80 +/- 64 microM (p = .003 compared with patients without criteria-based sepsis), respectively. The ability of plasma nitrite/nitrate > 54 microM to identify patients with physician diagnosed sepsis is characterized as follows: 87% sensitivity, 77% specificity, 50% positive predictive value, and 96% negative predictive value. The ability of plasma nitrite/nitrate > 54 microM to identify patients with criteria-based sepsis is characterized as follows: 61% sensitivity, 68% specificity, 21% positive predictive value, and 92% negative predictive value. CONCLUSIONS: Clinical diagnosis of SIRS is strongly associated with increased total plasma nitrite/nitrate concentrations in pediatric patients in the pediatric ICU. Many patients with increased nitrite/nitrate concentrations have inflammation without having a clinical diagnosis of SIRS. Our data suggest that increased plasma nitrite/nitrate concentrations are the standard for identifying patients with inflammation in the pediatric ICU. PMID- 9201065 TI - End-of-life issues: the physician's role. PMID- 9201066 TI - A "prone dependent" patient with severe adult respiratory distress syndrome. PMID- 9201067 TI - Intensive care unit costs. PMID- 9201068 TI - Role of positive end-expiratory pressure in extravascular lung water decrease. PMID- 9201069 TI - Irritable bowel syndrome. Diagnosis in the managed care era. AB - The common occurrence of irritable bowel syndrome underscores the importance of an accurate diagnostic evaluation without unnecessary expense. A preliminary diagnosis can usually be made with the Manning symptom criteria and additional history data in patients without warning signs of organic disease. A confident diagnosis can often be made with the addition of a physical examination and only limited laboratory and structural studies, such as a proctosigmoidoscopy and complete blood count. Tests that may be indicated in some patients include fecal examination for parasites and occult blood, dietary lactose exclusion or a lactose-hydrogen breath test, and a complete colon structural study. Other tests are occasionally indicated. Routine rectal biopsy and abdominal ultrasonography are unnecessary in patients with only typical symptoms, and large bowel motility testing is not useful. After a confident diagnosis, further testing for recurrent symptoms can be minimized. Investigation for psychosocial factors, while not necessary to diagnose irritable bowel syndrome, is important in treatment and may reduce medical costs. Misdiagnosis can result in unnecessary hysterectomy and other surgery, and it may be reduced by closer collaboration with gynecologists and general surgeons in the evaluation of patients. PMID- 9201070 TI - Sigmoid afferent mechanisms in patients with irritable bowel syndrome. AB - Up to 60% of patients with IBS have lowered perception thresholds in the rectum to balloon distension. The current study sought to test the hypothesis that IBS patients with normal perception thresholds in the rectum show hypersensitivity of afferent pathways in the sigmoid colon. Eleven healthy normal subjects and eight IBS patients with normal rectal perception thresholds underwent a balloon distension protocol in the sigmoid and rectum. Discomfort thresholds, receptive relaxation, compliance, and referral patterns were measured. Although IBS patients had significantly lower discomfort thresholds in the sigmoid when measured as volume, pressure, and wall tension, thresholds were similar to normals. Receptive relaxation and dynamic compliance were significantly decreased in IBS patients in the sigmoid. Referral patterns were similar during sigmoid distention in IBS patients in comparison to normals. Despite normal perception thresholds in rectum and sigmoid, IBS patients show evidence for alterations in rectosigmoid afferent mechanisms. In the sigmoid, this is seen in the form of reduced reflex relaxation and compliance and in the rectum in the form of altered viscerosomatic referral. PMID- 9201072 TI - Local and systemic complement activity in small intestinal bacterial overgrowth. AB - It is unknown whether bacteriolysis due to luminal complement activation contributes to local defense mechanisms against small intestinal bacterial overgrowth, particularly with gram-negative bacteria. This study addressed this issue. Thirty adult subjects were investigated with culture of luminal secretions adherent to proximal small intestinal mucosa. Luminal and plasma concentrations of C3 and C3d and C3d/C3 ratios were determined. Activated terminal complement complex was sought in surface epithelium to which aspirated secretions had been adherent. Small intestinal bacterial overgrowth with gram-negative bacteria was present in 12/30 (40.0%) subjects. C3, C3d, and C3d/C3 profile indicated that increased local but not systemic C3 activation occurs in this group. Conversely, no activation of terminal complement complex was evident in this circumstance. Thus, complement-mediated bacteriolysis is unlike to contribute to local defense mechanisms against small intestinal bacterial overgrowth, even when overgrowth flora includes gram-negative bacteria. Factors preventing full local activation of the complement cascade in this circumstance require investigation. PMID- 9201071 TI - Antisecretory mechanisms of peptide YY in rat distal colon. AB - Peptide YY (PYY) is a potent regulator of intestinal secretion. These studies investigated the role of Y1 and Y2 receptor subtypes in mediating the antisecretory effects of PYY on mucosa-submucosa preparations of rat distal colon. Addition of vasoactive intestinal peptide (VIP) to these tissues resulted in a 140 +/- 18% increase in basal short-circuit current (Isc) and the induction of Cl- secretion. VIP-stimulated increases in Isc were abolished by the addition of each of PYY, (Pro34)-PYY, a Y1 receptor-selective agonist, and PYY-(3-36), an endogenous Y2 receptor-selective ligand. However, when tissue neural transmission was blocked with tetrodotoxin, neither PYY nor its receptor subtype-selective analogs were able to inhibit VIP-stimulated increases in Isc. These results suggest that in the rat distal colon, the antisecretory actions of PYY are mediated through a combination of Y1 and Y2 receptor subtypes or through a novel receptor subtype that is unable to discriminate between (Pro34)-PYY and PYY-(3 36). PMID- 9201073 TI - Serum n3 polyunsaturated fatty acids are depleted in Crohn's disease. AB - To determine fatty acid patterns in Crohn's disease, we measured various serum fatty acids by gas chromatography in 20 patients with the disease and compared them with those in 18 healthy controls. All the patients had been free from any nutritional supplementation during preceding six months or had no history of intestinal resection. Eight of the patients were affected in the small bowel only, three in the large bowel only, and the remaining nine in both the small and large bowel. Both serum concentrations and percentages of C20:4n6, C20:5n3, C22:0, C22:6n3, total n3 polyunsaturated fatty acids, and total polyunsaturated fatty acids were lower in the patients than in the controls. Both essential fatty acids (C18:2n6, C18:3n3) and C20:3n9 levels were not different between the two groups. Among nine fatty acids that correlated with the Crohn's disease activity index, C20:5n3 and total n3 polyunsaturated fatty acids showed the most significant negative correlations. These findings suggest that essential fatty acid deficiency rarely occurs in Crohn's disease and also that n3 polyunsaturated fatty acids may be relevant to the activity of the disease. PMID- 9201074 TI - Inflammatory myositis in association with inflammatory bowel disease. PMID- 9201076 TI - Comparative assessment of power dynamics of gastric electrical activity. AB - This study was undertaken to investigate the correlation between power dynamics of gastric electrical activity (GEA) assessed with different recording techniques. A total of seven eight-channel 1-hr combined recordings were obtained from three subjects in five consecutive postoperative days. Four channels were recorded from bipolar electrodes implanted into the gastric antral wall, and four channels were electrogastrographic (EGG). Six pairs of bipolar electrodes were inserted into the antral wall (three anterior; three posterior) of 16 anaesthetized dogs. Fourteen-channel (six internal GEA and eight EGG) 1/2-hr recordings were obtained from each dog. Sets of power values calculated from channel pairs (internal, EGG or mixed) were cross-correlated and the significance of the obtained correlation coefficients was examined (P < 0.05). The majority of power correlations of internal GEA channel pairs, and those of mixed (internal GEA-EGG) channel pairs were insignificant. These findings question the claims that EGG power dynamics mirrors the power dynamics of internal GEA. PMID- 9201075 TI - Distribution of beta-adrenoceptor subtypes in gastrointestinal tract of nondiabetic and diabetic BB rats. A longitudinal study. AB - The effects of aging and diabetes on the distribution of beta-adrenoceptor subtypes in the gut were investigated in the BB rat. [125I]Cyanopindolol binding to 10-micron sections was evaluated using film autoradiography. Cyanopindolol binding to beta-, beta 1-, and beta 2-adrenoceptors was displaced by 1 microM propranolol, 50 nM ICI-89-406, and 100 nM ICI-118-551, respectively. beta Adrenoceptor binding was highest in the circular muscle of proximal colon and lowest in the pylorus of 4- to 5-month-old rats. Aging (8- to 10-month-old vs. 4- to 5-month-old rats) was associated with increased beta-adrenoceptor binding in the pylorus and reduced binding in the proximal colon. Diabetes had a time dependent effect on the level of beta-adrenoceptor binding. It was increased in the antral and pyloric stomach but longer periods of diabetes caused a reduction in beta-adrenoceptor binding in the pylorus. Those in the intestine were reduced time-dependently and involved beta 1- or beta 2-adrenoceptors or both. PMID- 9201077 TI - Gastric emptying rate of solids in patients with nonulcer dyspepsia. AB - The underlying role of motility disorders and delayed gastric emptying in nonulcer dyspepsia is still questioned. This study aimed to determine the role of the gastric emptying rate of solids in patients with nonulcer dyspepsia. By means of breath test technology, gastric emptying results of 344 consecutive patients with nonulcer dyspepsia were compared with those of 70 normal healthy volunteers. Although gastric emptying was significantly delayed in patients with nonulcer dyspepsia compared with normal volunteers, there was a great overlap between the two groups. Using 5-95% confidence intervals of the control group in about 30% of the patients with nonulcer dyspepsia gastric emptying was delayed. No correlation was found between gastric emptying rate and age, weight, height, or sex of the subjects in both groups. These findings suggest that, apart from gastric emptying, other mechanisms are very important in the etiology of nonulcer dyspepsia. PMID- 9201078 TI - Chronic intestinal pseudoobstruction associated with fetal alcohol syndrome. AB - Alcohol acts as a teratogen in the fetus, resulting in prenatal or postnatal growth failure, characteristic facial dysmorphic features, and central nervous system dysfunction. The toxic effects of alcohol on the developing brain are well recognized, but gastrointestinal neuropathy has not been described in fetal alcohol syndrome (FAS). Five children with FAS presented in infancy with signs and symptoms suggestive of chronic intestinal pseudoobstruction. They were not able to sustain adequate caloric intake by mouth, and all required prolonged special methods of alimentation. We performed antroduodenal manometry in these children to determine whether their symptoms were associated with a gastrointestinal motility disorder. All patients had abnormally propagating phase III-like episodes during fasting (retrograde in four, simultaneous in two). Persistent clusters of stationary contractions were a prominent feature in two patients. In utero neurotoxicity of alcohol may not be limited to the central nervous system, but may also cause an enteric neuropathy presenting in infancy as chronic intestinal pseudoobstruction. PMID- 9201079 TI - Effect of octreotide on human sphincter of Oddi motility following liver transplantation. AB - The effect of octreotide on sphincter of Oddi motility was investigated in six liver transplant patients, employing percutaneous (through the T-tube tract) manometry. Continuous and simultaneous sphincter of Oddi and duodenal motor activities were recorded before and for 60 min after the administration of octreotide (100 micrograms subcutaneously) and after the injection of cholecystokinin (0.02 microgram/kg intravenously). With octreotide, contraction frequency and basal pressure significantly increased (P < 0.05). This effect lasted more than 60 min, long after octreotide-induced duodenal migrating motor complex phase III activity had ceased. Sphincter of Oddi contraction amplitude and duration were unaffected by octreotide. Subsequent cholecystokinin administration transiently reduced sphincter of Oddi basal pressure and contraction frequency. We conclude that octreotide significantly increases sphincter of Oddi basal pressure and contraction frequency. This effect is distinct from octreotide induction of migrating motor complex phase III activity, persists for a prolonged period, and is inhibited by cholecystokinin. PMID- 9201080 TI - Medical residents' colorectal cancer screening may be dependent on ambulatory care education. AB - Colorectal cancer results in significant morbidity and mortality in the United States. Screening is a critical component of cancer prevention. However, research has suggested that physicians may inconsistently adhere to surveillance guidelines. Since residency training can significantly impact upon future practice patterns, assessment of postgraduate colorectal cancer education is important. This retrospective chart review of patients > or = 50 years of age compared screening performed by resident physicians' in different internal medicine residency programs at The George Washington University Medical Center. Resident physicians who received multiple lectures in colorectal cancer surveillance or were required to document performance of screening on a medical record preventive care summary form performed significantly more rectal examinations (P < 0.0004), fecal occult blood testing (P < 0.00001), and flexible sigmoidoscopies (P < 0.00001) when compared to other resident physicians. Postgraduate education should employ multiple education techniques and reinforcement procedures to increase physician compliance with cancer screening. PMID- 9201081 TI - Jejunal lymphangioma. An unusual cause of chronic gastrointestinal bleeding. AB - We discuss a case of lymphangioma of the small bowel that caused chronic anemia secondary to gastrointestinal blood loss. We believe that this is the first jejunal lymphangioma to be diagnosed by Sonde endoscopy. There are several other lymphatic abnormalities of the small intestine that are briefly discussed. In the Western literature, anemia or chronic gastrointestinal blood loss is an unusual presentation of lymphangioma of the small bowel. PMID- 9201082 TI - Esophageal dysmotility and gastroesophageal reflux in intrinsic asthma. AB - This study was undertaken to determine the prevalence of esophageal motor abnormalities, the incidence of gastroesophageal reflux, and the coexistence of gastroesophageal reflux with esophageal dysmotility in patients with intrinsic asthma. Based on clinical criteria, 34 consecutive asthmatics, 15 patients with gastroesophageal reflux, and 10 subjects with upper gastrointestinal symptoms with normal results of esophageal manometry and 24-hr esophageal pH test (controls) were studied. Esophageal motor disorders were noted in 23 of 34 asthmatics, and in 10 of 15 patients with acid reflux but in none of the subjects of the control group. A positive result of the prolonged esophageal pH study (pH in the distal esophagus less than 4 for more than 4.2% of the recording time) was obtained in 14 of 17 patients with asthma (only 17 of the original patients were tested because the others did not give informed consence for this test) and in all patients with gastroesophageal reflux. None of the members of the control group had positive test results. The findings of this study show that: (1) it is possible to identify a group of subjects with nonallergic asthma presenting with esophageal dysmotility, (2) the 24-hr esophageal pH study must be properly done in such patients; (3) esophageal motor abnormalities are often associated with positive pH results; and (4) more reflux was observed while in a supine position (especially during the night) than that observed either in control or reflux patients. Based on these results, patients with intrinsic asthma with reflux can benefit from both acid suppressive and prokinetic drugs with notable clinical implications regarding standard treatment for asthma, and those with prevalent supine compared to upright reflux could even benefit from surgery. PMID- 9201083 TI - Effects of red wine on 24-hour esophageal pH and pressures in healthy volunteers. AB - The purpose of this study was to assess the effects of red wine taken with meals on esophageal motility, esophageal exposure to acid, and gastric pH. Following a randomized design, 14 healthy male volunteers (mean age 25 years, range 18-35 years were given 360 ml of red wine or tap water during lunch or dinner. All subjects underwent ambulatory 24-hr esophageal motility and esophagogastric pH monitoring studies. Three different periods were analyzed: during meals (30 min), postprandial (3 hr), and 8-hr supine. Two volunteers complained of heartburn after wine ingestion. An increase in the number of high amplitude waves (> 125 mm Hg, 95th percentile of our motility unit controls) was observed during meals accompanied by wine: water 1.2 (0-10.2), wine 1.6 (0-32.6), P = 0.02 [median (range)]. No other esophageal motility changes occurred. Percent reflux time increased during the postprandial period after wine ingestion in comparison with water: 1.7 (0-14.9) vs 0.1 (0-0.8), P < 0.05. Gastric pH was unaffected by the type of drink. Ingestion of moderate amounts of red wine with meals increases postprandial esophageal exposure to gastric acid in healthy persons. PMID- 9201084 TI - Inhibition of parietal cell acid secretion is mediated by the classical epidermal growth factor receptor. AB - Epidermal growth factor (EGF) and transforming growth factor-alpha (TGF-alpha) inhibit gastric acid secretion both in vivo and in vitro. Previous studies have indicated that EGF and TGF-alpha bind to the same EGF/TGF-alpha receptor. Nevertheless, we and others have previously demonstrated that inhibition of acid secretion by these growth factors requires concentrations of the peptides that are 10-fold higher than those necessary for induction of mitogenesis. Therefore, we have sought to investigate whether gastric parietal cells may possess a second EGF/TGF-alpha receptor class. Two systems were studied: First, [125I]TGF-alpha was cross-linked to the receptor in isolated rabbit parietal cell membranes, and labeled species were resolved on SDS-PAGE. Second, acid secretion was evaluated in pylorus-ligated waved-2 mutant mice, which carry a disabling point mutation in their classical EGF/TGF-alpha receptor. In isolated parietal cells, [125I]TGF alpha was cross-linked into a single species of 170 kDa. Cross-linking was inhibited in the presence of unlabeled TGF-alpha with an IC50 of 80 nM. In the pylorus-ligated mice, control littermate mice demonstrated a dose-dependent inhibition of acid secretion by EGF with an IC50 of 20 micrograms/kg. In contrast, EGF had no inhibitory effect on acid secretion in waved-2 mice at concentrations up to 100 micrograms/kg. No alterations in parietal cell or gastrin cell numbers were observed. These results in both isolated rabbit parietal cells and waved-2 mice support the existence of only a single class of EGF/TGF-alpha receptors in parietal cells. Differences in growth factor affinity are likely due to the modification of the receptor or one of its coordinate regulators. PMID- 9201085 TI - Immunohistochemical studies on EGF family growth factors in normal and ulcerated human gastric mucosa. AB - Expression of members of the epidermal growth factor family, including epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), amphiregulin (AR), and Cripto, as well as their putative receptor, epidermal growth factor receptor (EGFR), was studied immunohistochemically in human gastric mucosa to evaluate their possible roles in cell proliferation of normal and regenerative gastric mucosa. We also examined the correlation betwen cell proliferation and EGFR by double immunohistochemical staining for proliferating cell nuclear antigen (PCNA) and EGFR. In normal gastric mucosa, TGF-alpha, Cripto, and AR immunoreactivities were observed in the surface epithelial and parietal cells of gastric fundic glands, respectively. EGF immunoreactivity was not observed in any of normal mucosa examined. EGFR immunoreactivity was detected on foveolar cells in proliferative zones and in parietal cells. Double immunostaining revealed that EGFR immunoreactivity was distributed much more widely than PCNA immunoreactivity. PCNA positive epithelial cells adjacent to gastric ulcer margin expressed relatively intense EGFR but did not express any of the growth factors examined. On the other hand, relatively intense immunoreactivity of both TGF alpha and Cripto was detected in PCNA-negative regenerative epithelium located distant from gastric ulcer margin. Relative immunoreactivity of AR in regenerative gastric epithelium associated with ulcer was not different from that in normal gastric mucosa. TGF-alpha, AR, and Cripto are considered to play important roles in normal gastric mucosal proliferation, and TGF-alpha and Cripto may be involved in ulcer healing, possibly via a paracrine mechanism. PMID- 9201086 TI - Interleukin-8 stimulates leukocyte migration across a monolayer of cultured rabbit gastric epithelial cells. Effect associated with the impairment of gastric epithelial barrier function. AB - Acute Helicobacter pylori infection produces predominantly neutrophilic infiltration of the gastric mucosa. However, the precise mechanisms and mediators of neutrophil migration are not known. Interleukin-8 (IL-8), a potent chemotactic factor for neutrophils, is present at high concentration in the gastric mucosa of subjects with chronic gastritis caused by H. pylori infection. The aims of this study were to determine whether IL-8 stimulates polymorphonuclear leukocyte (PMN) migration across a cultured monolayer of rabbit gastric epithelial cells and whether PMN migration affects epithelial cell barrier function. Confluent gastric epithelial monolayers grown on the inserts were overlaid with PMNs and various amounts of IL-8 were administered into the well under the insert. Gastric epithelial barrier function was assessed by sodium back diffusion. IL-8 stimulated PMN migration across the monolayer in a dose- and time-dependent manner. PMN transmigration significantly increased sodium back diffusion. In conclusion, IL-8 induces PMN migration across a monolayer of cultured gastric epithelial cells. This IL-8 action is associated with impairment of gastric epithelial barrier function. Since H. pylori infection causes a local mucosal increase of IL-8, our present findings may explain the mechanism of H. pylori induced PMN infiltration of the gastric glands and mucosal injury. PMID- 9201087 TI - Effects of adrenalectomy on serotonin-, somatostatin-, and gastrin-immunoreactive cells in rat gastrointestinal tract. AB - The effects of bilateral adrenalectomy on the serotonin-, somatostatin-, and gastrin-immunoreactive cells in the rat gastrointestinal tract were studied four weeks after surgery. Body weight was reduced and the small intestine shorter in adrenalectomized animals compared with controls, while no changes were found in the histology of the mucosa. In the adrenalectomized animals the number of serotonin-immunoreactive cells was increased in the cecum and large intestine, while the somatostatin-immunoreactive cells were decreased in number in the antrum and increased in the corpus, cecum, and large intestine. The gastrin immunoreactive cells in the antrum were not affected in number, but their nuclear size was enlarged, possibly indicating increased cellular activity. PMID- 9201088 TI - Relationship between changes of active oxygen metabolism and blood flow and formation, progression, and recovery of lesions is gastric mucosa of rats with a single treatment of compound 48/80, a mast cell degranulator. AB - The relationship between the changes of active oxygen metabolism and blood flow and the formation, progression, and recovery of lesions was examined in the gastric mucosa of rats treated once with compound 48/80, a mast cell degranulator. Gastric mucosal lesions appeared 0.5 hr after compound 48/80 treatment, became worst at 3 hr, and recovered fairly well at 12 hr. Increases in gastric mucosal lipid peroxide content and xanthine oxidase and myeloperoxidase activities and decreases in gastric mucosal vitamin E and hexosamine contents and Se-dependent glutathione peroxidase activity occurred with the formation and progression of gastric mucosal lesions. These changes were attenuated with the recovery of the lesion. Gastric mucosal nonprotein SH content decreased with the formation of gastric mucosal lesions, and this decreased SH content returned to near the original level with lesion progression. No changes in gastric mucosal superoxide dismutase and catalase activities occurred with the formation, progression, and recovery of gastric mucosal lesions. Gastric mucosal blood flow decreased with the formation of gastric mucosal lesions, and this decreased blood flow recovered with lesion progression. Serum serotonin concentration, an index of mast cell degranulation, increased with the formation of gastric mucosal lesions, and this increased serotonin level was attenuated with lesion progression and recovery. Pretreatment with ketotifen, a connective tissue mast cell stabilizer, prevented the formation of gastric mucosal lesions, the increases of gastric mucosal lipid peroxide content, xanthine oxidase and myeloperoxidase activities, and serum serotonin level; and the decreases of gastric mucosal nonprotein SH content, glutathione peroxidase activity, and blood flow found at 0.5 hr after compound 48/80 treatment. These results indicate that the changes of gastric mucosal active oxygen metabolism and blood flow are closely related to the formation, progression, and recovery of gastric mucosal lesions in rats with a single compound 48/80 treatment. The present results also suggest that this compound 48/80-induced gastric mucosal injury could be a kind of ischemia-reperfusion-induced injury occurring through degranulation of connective tissue mast cells. PMID- 9201089 TI - Suramin enhances ethanol-induced injury to gastric mucosa in rats. AB - Suramin is currently used in clinical practice as antineoplastic agent because of its complex interaction with the biological activity of various growth factors involved in tumor progression. The influence exerted by suramin on gastric injury induced in rats by intraluminal injection of absolute ethanol was investigated in the present study. The morphometric analysis of gastric histological sections revealed that suramin, 18 mg/kg, administered intraperitoneally for 14 days every other day, caused a marked enhancement of ethanol-induced mucosal damage. This effect was more pronounced 1-8 hr following ethanol administration, and it was still significant after 48 hr. In suramin-treated animals the evaluation of Alcian blue recovery from gastric-bound mucus showed that the levels of adherent mucus were significantly lower than those detected in untreated rats. In addition, pretreatment with suramin did not modify basal acid secretion, but caused potentiation of acid output stimulated by pylorus ligation or electrical vagal stimulation. Overall, the present results indicate that suramin exerts a negative influence on both gastric protective and repairing mechanisms. Due to the peculiar pharmacodynamic profile of suramin, it is suggested that interference with endogenous growth factors, endowed with physiological protective activity on gastric mucosa, might account for the damage-enhancing action of this drug. PMID- 9201090 TI - Effects of several denervation procedures on distribution of calcitonin gene related peptide and substance P immunoreactive in rat stomach. AB - The effect of chemical deafferentation, vagotomy (VGX), and gangliosympathectomy (GSX) on the density of fibers containing calcitonin gene-related peptide (CGRP) and substance P (Sub.P) in the rat gastric wall was studied. Chemical deafferentation by capsaicin abolished the density of CGRP-immunoreactive (IR) fibers, not Sub.P-IR fibers. Ten days after VGX, the density of CGRP-IR or Sub.P IR fibers in the mucosa was largely reduced, while no reduction of CGRP-IR and Sub.P-IR fibers was seen in submucosal and muscular layers. GSX significantly reduced the density of CGRP-IR fibers in the mucosa and caused a moderate decrease in the fibers in submucosal and muscular layers. Pretreatment with 6 hydroxydopamine, a neurotoxin for noradrenergic nerves, did not affect the density of CGRP-IR fibers in the gastric wall. The density of Sub.P-IR fibers in the gastric wall was not affected by GSX. These studies indicate that the CGRP-IR and Sub.P-IR fibers in the mucosa are susceptible to extrinsic nerve denervation compared with those in the submucosa and muscle layers, that a major portion of the CGRP-IR fibers in the mucosa is of both vagal and spinal origin, and that a major portion of the Sub.P-IR fibers in the mucosa is of vagal origin. Furthermore, the present results support that CGRP-IR fibers, not Sub.P-IR fibers, in the rat stomach are capsaicin-sensitive. PMID- 9201091 TI - Comparison of omeprazole and ranitidine for stress ulcer prophylaxis. AB - Stress ulcer prophylaxis protects against clinically important gastrointestinal bleeding and has gained widespread use. This study compares the efficacy of omeprazole to ranitidine for this indication. This was a prospective, randomized clinical trial. Sixty-seven high-risk patients were randomized to receive either ranitidine 150 mg (N = 35) intravenously daily or omeprazole 40 mg (N = 32) daily orally or by nasogastric route. Patients were monitored for clinically important bleeding. There was no statistically significant difference between treatment groups in the number of patients enrolled, gender, race, or age. The study groups were comparable in regard to the severity of illness based on their similar APACHE II score, duration of ICU stay, duration of ventilator dependence, and mortality rate. A significant difference was found only in regard to the number of risk factors per patient. The ranitidine-treated group had 2.7 risk factors per patient while the omeprazole-treated group had 1.9 (P < 0.05). Eleven patients (31%) given ranitidine and two patients (6%) given omeprazole developed clinically important bleeding (P < 0.05). Nosocomial pneumonia developed in five patients (14%) receiving ranitidine and one patient (3%) receiving omeprazole (P > 0.05). We conclude that oral omeprazole is safe, effective, and clinically feasible for stress ulcer prophylaxis. PMID- 9201092 TI - Alteration of dopamine D2 receptors in human malignant stomach tissue. AB - Dopamine is an important enteric neurotransmitter with a wide spectrum of physiological actions on the gastrointestinal tract. In addition, it showed inhibition of malignant cell proliferation as well as a protective influence on experimental carcinogenesis in the gastrointestinal tract of murine hosts. It is well established that dopamine acts on target cells through specific receptors. Therefore the status of dopamine receptors in malignant tumors of the stomach has been evaluated. Normal, benign, and malignant stomach tissue showed the presence of high-affinity D2 dopamine receptors. The concentration (Bmax) and affinity (Kd) of dopamine binding sites in normal and benign tumor tissues were similar. In malignant stomach tissue Bmax showed a significant decrease compared to normal and benign controls; however, Kd was similar. This alteration of dopamine receptors may be of significance in understanding the etiopathogenesis of gastric cancer at the level of peripheral neurotransmitters. Rational use of dopamine receptor antagonists for various stomach diseases may be suggested. PMID- 9201093 TI - Prognostic indicators for survival after curative resection for patients with carcinoma of the stomach. AB - This study aims to determine prognostic indicators among patient-, tumor-, and treatment-related factors of gastric cancer patients. A total of 510 patients who underwent curative gastric resection were studied. Univariate analysis of patient related factors showed a significantly lower survival in patients with a history of obstruction, hypoalbuminemia, and anemia. Tumor-related factors including gross appearance, location, and size of tumor; depth of cancer invasion; level, number, and frequency of lymph node metastasis; stromal reaction and tumor growth pattern; and histological classification all significantly affected survival. Surgical treatment related factors such as total or distal subtotal gastrectomy, extent of lymphadenectomy, and combined resection of adjacent organ(s) showed a statistically significant adverse influence on survival. Multivariate analysis identified only four tumor-related factors-number of metastatic lymph nodes, depth of cancer invasion, stromal reaction, and gross appearance of the tumor-as independently affecting survival. These findings suggest that only four tumor related factors were prognostic indicators in patients with gastric cancer. PMID- 9201094 TI - Peritoneal mucinous carcinomatosis. A possible explanation for an unusual laparoscopic appearance. PMID- 9201096 TI - Activation of human neutrophils by calcium carbonate polymorphs. AB - Gallstone formation is frequently accompanied by inflammation of the gallbladder mucosa. Some gallstone components such as cholesterol, calcium bilirubinate, and calcium hydroxyapatite have been previously shown to activate neutrophils. We investigated the effect on neutrophils of the calcium carbonate polymorphs aragonite, calcite, and vaterite (all found in gallstones). By chemiluminescence, superoxide, and degranulation assay, all three crystals were shown to cause rapid activation of neutrophils. The potency of the crystals was aragonite > vaterite > calcite. In vivo, crystals may be plasma-protein-coated before they encounter neutrophils; therefore some experiments were repeated using crystals that had been preincubated with plasma. For aragonite and vaterite, protein adsorption decreased the chemiluminescence response by approximately 50%. In contrast, protein-coated calcite crystals elicited a greater chemiluminescence response than did uncoated crystals. In summary, the calcium carbonate polymorphs are potent activators of neutrophils and thus have the potential to contribute to gallstone-associated cholecystitis. PMID- 9201095 TI - Successful topical dissolution of cholesterol gallbladder stones using ethyl propionate. AB - Topical dissolution of cholesterol gallbladder stones using methyl tert-butyl ether (MTBE) is useful in symptomatic patients judged too ill for surgery. Previous studies showed that ethyl propionate (EP), a C5 ester, dissolves cholesterol gallstones rapidly in vitro, but differs from MTBE in being eliminated so rapidly by the liver that blood levels remain undetectable. Our aim was to test EP as a topical dissolution agent for cholesterol gallbladder stones. Five high-risk patients underwent topical dissolution of gallbladder stones by EP. In three patients, the solvent was instilled via a cholecystostomy tube placed previously to treat acute cholecystitis; in two patients, a percutaneous transhepatic catheter was placed in the gallbladder electively. Gallstone dissolution was assessed by chromatography, by gravimetry, and by catheter cholecystography. Total dissolution of gallstones was obtained in four patients after 6-10 hr of lavage; in the fifth patient, partial gallstone dissolution facilitated basketing of the stones. In two patients, cholesterol dissolution was measured and averaged 30 mg/min. Side effects were limited to one episode of transient hypotension and pain at the infusion site; no patient developed somnolence or nausea. Gallstone elimination was associated with relief of symptoms. EP is an acceptable alternative to MTBE for topical dissolution of cholesterol gallbladder stones in high-risk patients. The lower volatility and rapid hepatic extraction of EP suggest that it may be preferable to MTBE in this investigational procedure. PMID- 9201097 TI - Biliary interleukin-6 and tumor necrosis factor-alpha in patients undergoing endoscopic retrograde cholangiopancreatography. AB - Cytokines are low-molecular-weight protein mediators that possess a wide spectrum of inflammatory, metabolic, and immunomodulatory properties. Cytokines have been shown to be produced by monocytes/macrophages, lymphocytes, fibroblasts, endothelial cells, and more recently, hepatocytes and biliary epithelium. The aim of this study was to define biliary levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) in various disease states. Fifty-four patients undergoing ERCP comprised the study group. IL-6 and TNF-alpha were measured in aspirated bile using an ELISA technique. Levels of both TNF-alpha and IL-6 were significantly higher in patients with cholangitis (P < 0.00001). Moreover, IL-6 was 100% specific for cholangitis since none of the patients without bacterial cholangitis-including patients with biliary obstruction secondary to cholangiocarcinoma or pancreatic carcinoma-had measurable IL-6 in their bile. Low levels of biliary TNF-alpha were detectable in five patients without cholangitis; the sensitivity and specificity of TNF-alpha for cholangitis were 100% and 82%, respectively. There was a strong statistical correlation between biliary IL-6 and TNF-alpha levels (r = 0.819, P < 0.0001). In contrast, the correlations between biliary cytokines and serum biochemical parameters were weak. These results suggest that IL-6 and TNF-alpha are sensitive markers for cholangitis and may differentiate it from other types of biliary tract disease. PMID- 9201098 TI - Diffuse intrahepatic biliary strictures in sarcoidosis resembling sclerosing cholangitis. Case report and review of the literature. AB - We report a case of sarcoidosis with severe cholestasis and cholangiographic features of sclerosing cholangitis that responded dramatically to corticosteroid therapy. Although an association between sarcoidosis and primary sclerosing cholangitis has been suggested by previous reports, features suggestive of primary sclerosing cholangitis, including inflammatory bowel disease, hepatic histology and serum neutrophil cytoplasmic antibodies, were absent in this case. Cholangiography may be useful in the evaluation of patients with cholestatic sarcoid liver disease, and intrahepatic biliary strictures should be included in the spectrum of hepatic involvement by sarcoidosis. A trial of corticosteroid therapy may be of benefit in patients with bile ductal involvement by sarcoidosis. PMID- 9201100 TI - Predictive value of family history in diagnosis of hereditary hemochromatosis. AB - Our objective was to study the predictive value of the family history in the initial diagnosis of hereditary hemochromatosis. Sixty five hemochromatosis proband patients and 66 control patients with chronic liver disease were assessed for a family history of hemochromatosis, cirrhosis, diabetes, and arthritis. There were no significant differences in the frequency of cirrhosis, diabetes, and arthritis between hemochromatosis patients and control patients. A family history of hemochromatosis was present in 3.6% of hemochromatosis patients and none of the control patients. Multivariate discriminative analysis demonstrated that the combination of cirrhosis, diabetes, and arthritis could only predict the diagnosis of hemochromatosis in 48% of cases. We conclude that a family history of cirrhosis, arthritis, and diabetes is not more common in hemochromatosis patients compared to control patients with chronic liver disease. PMID- 9201099 TI - Method of measurement of pancreatic elastase II activity and postnatal development of proteases in human duodenal juice and bovine and porcine pancreatic tissue. AB - A specific method for pancreatic elastase II activity analysis was developed. True elastase II activity could be discriminated from that of elastase I and chymotrypsin. The postnatal development of four pancreatic proteases in the duodenal juice of children and in the pancreatic homogenates of calves and piglets was measured. The study was carried out on patients without (14 children) and with (5 children) pancreatic insufficiency. Calves and piglets were either milk-fed or weaned until slaughter at different ages. Profiles of enzyme development were globally similar in milk-fed piglets and calves, while in children without pancreatic insufficiency, no significant change was observed between 4 and 168 months. In children with pancreatic insufficiency, enzyme activity was low. In animals, elastase II and chymotrypsin activities were maximal at birth, decreased with age, and probably were associated with the digestion of milk protein. In contrast, elastase I and trypsin activities increased markedly after weaning in connection with the intake of solid food. PMID- 9201101 TI - Effects of mixed ETA and ETB-receptor antagonist (Ro-47-0203) on hepatic microcirculation after warm ischemia. AB - There is evidence that endothelin (ET) is involved in disturbances of the hepatic microcirculation after warm ischemia. In this study we investigated the influence of a mixed ETA-, ETB-receptor antagonist (Bosentan) on ischemia-reperfusion damage of the liver by means of intravital fluorescence microscopy (IVM). Clamping of the left liver lobe (= warm ischemia) was performed in 16 male Wistar rats for 70 min. The treatment group (N = 8) received 15 mg/kg Bosentan (Ro-47 0203) 1 min prior to reperfusion. Controls (N = 8) received an equivalent amount of Ringer's solution. Between 20 and 90 min after reperfusion, leukocyte endothelial cell interactions in sinusoids and postsinusoidal venules as well as perfusion of hepatic acini were studied. Application of Bosentan improved sinusoidal blood flow, attenuated manifestations of microvascular perfusion failure, and decreased the number of rolling leukocytes in postsinusoidal venules. Our results provide further evidence that ET is involved in postischemic impairment of hepatic microhemodynamics during reperfusion. PMID- 9201103 TI - Dobutamine stress echocardiography for the detection of coronary artery disease and viable myocardium. AB - Dobutamine stress echocardiography has become a diagnostic tool for the evaluation of coronary artery disease and the detection of myocardial viability. In the diagnosis of significant coronary artery disease, it provides similar accuracy to exercise stress thallium-201 myocardial perfusion scintigraphy. Dobutamine stress echocardiography is also a promising modality for predicting the recovery of hibernating myocardium from contractile dysfunction after coronary angioplasty or bypass surgery. This article, reviews our recent clinical experience with the detection of coronary artery disease and viable myocardium by dobutamine stress echocardiography. PMID- 9201102 TI - Effect of dietary nucleotides on degree of fibrosis and steatosis induced by oral intake of thioacetamide. AB - The administration of thioacetamide in rats induces nodular cirrhosis of the liver, characterized by fibrous septae, parenchymal nodules, proliferation of the bile ducts, and excessive deposition of connective tissue elements. Nodular cirrhosis is also associated with changes in lipid metabolism, as shown by the accumulation of lipid droplets in the hepatocyte cytoplasm. Adequate nutritional support during cirrhosis is important to sustain liver function and promote recovery after the lesions have been induced. Supplementation with nucleotides may increase cellular proliferation and thus optimize hepatic recovery. The aim of this study was to investigate the effects of dietary nucleotide supplementation on the degree of fibrosis and steatosis in rats with liver cirrhosis induced by four months of oral intake of thioacetamide. The use of dietary nucleotides after thioacetamide administration was found to decrease the percentage area of fibrous septae. In animals with liver cirrhosis fed the nucleotide-supplemented diet for two weeks, the total area of fibrosis was reduced. Withdrawal of the hepatotoxic agent led to a decrease in the degree of steatosis in cirrhotic animals, which was significant in rats given the nucleotide-supplemented diet during a two-week recovery period. In conclusion, dietary nucleotides may be an important factor in the histological recovery of damaged liver in experimental cirrhosis. PMID- 9201104 TI - Cardiac involvement in progressive muscular dystrophy of the Duchenne type. AB - Duchenne's progressive muscular dystrophy (DMD) is a genetic muscle disorder that causes degeneration and atrophy of the systemic and cardiac muscle. The disease is manifested early in childhood, and most of patients die by age 20 years of respiratory failure or heart failure. The cardiac involvement in DMD is characterized pathologically by degeneration and fibrosis of the myocardium, centering around the posterolateral wall of the left ventricle. Functionally, an abnormal electrocardiogram, valve motion, wall thickness, and wall motion are observed. Furthermore, abnormalities in plasma levels of atrial natriuretic peptide and autonomic function are also demonstrated. In this review, the cardiac involvements in DMD in the following aspects are described: 1) Electrocardiogram; a) high-frequency notches on the QRS complexes, b) amplitude of QRS complexes, c) late potential, d) arrhythmias, e) heart rate variability, f) a 10-year follow-up study, 2) Echocardiographic findings, 3) Hemodynamic findings, 4) Atrial natriuretic peptide. PMID- 9201105 TI - Long-term outcome in double-vessel coronary artery disease in Japanese patients. AB - The long-term (average: 10 years) outcome in 220 patients with double-vessel disease (DVD) treated medically was investigated. The patients underwent coronary angiography between September 1973 and February 1984, and significant (75% or more) stenosis was detected in each of two major coronary arteries. These patients showed relatively good 5-year and 10-year survival rates of 94.5% and 87.4%, respectively. Cardiac death occurred in 31 patients (14.1%) and nonfatal myocardial infarction (MI) developed in 16 patients (7.3%) during follow-up. When these were defined as cardiac events, the annual attrition rate was 3.1%. A comparison of the outcome with regard to the presence or absence of MI revealed worse results for the MI group, but no difference was observed between different sites of infarction. There was also no difference in outcome with regard to the presence or absence of lesions in the left anterior descending artery (LAD). In the MI group, patients with impaired left ventricular function (ejection fraction < or = 40%) had inferior survival to those with good left ventricular function. Thus, DVD associated with good left ventricular function had a relatively good outcome when treated medically, while patients with impaired left ventricular function might benefit from revascularization. PMID- 9201106 TI - Patency of intermediate size side branches after Palmaz-Schatz stent implantation. AB - The immediate and long-term patency of intermediate size side branches was assessed by serial coronary angiography in 47 patients with 48 lesions to determine whether the presence of these side branches (1-2 mm in diameter) is unsuitable for Palmaz-Schatz stent implantation. Coronary angiography was performed at baseline, after conventional balloon angioplasty, immediately after stent implantation, at 1 week, and at 6-month follow-up. Sixty-eight lesion associated side branches that were 1-2 mm in diameter, 11 with branch ostial stenosis (Group A) and 57 without ostial stenosis (Group B) were studied. After stent implantation, 9 (13%) branches became totally occluded and coronary flow deteriorated in 13 branches (19%). The incidence of side branch occlusion during the procedure in group A was greater than in group B (55% vs. 12%; p < 0.005). One (2%) patient suffered persistent chest pain, but no procedure was complicated by Q-wave myocardial infarction or significant elevation of creatine kinase concentration. Flow improved in 82% of the occluded side branches after 1 week and in 90% after 6 months. These results suggest that the presence of intermediate size side branches is not a contra-indication to Palmaz-Schatz stent implantation. PMID- 9201107 TI - Transesophageal echocardiography in various ischemic stroke subtypes. AB - Transesophageal echocardiography (TEE) is more sensitive than transthoracic echocardiography (TTE) in detecting the potential source of emboli in cardioembolic strokes (CES). To establish the prevalence of a potential cardiac source of embolism detectable on TEE and its relationship to vascular risk factors, an unselected ischemic stroke population was evaluated. Twenty-six age and sex-matched cases with normal cardiological and neurological examinations as well as normal CT-scans, TTE and ECGs were included in the study as the control group. One hundred and eight patients with cardioembolic stroke (53 patients), atherothrombotic stroke (36 patients), and lacunar stroke (19 patients) were investigated by TTE and TEE. Seven of the 26 (26.9%) controls had thoracic atherosclerotic plaques on TEE examinations. The prevalence of abnormal TEE findings in patients was higher compared to the controls (p < 0.001). TEE revealed more specific findings in every etiological group when compared to TTE (74.0% vs 10.2%, p < 0.001). Atrial fibrillation correlated with the abnormalities of TEE (p < 0.05) while other risk factors did not. Left atrial spontaneous echo contrast was the most common finding on TEE of cases with cardioembolic stroke while atherosclerotic aortic plaques were mostly encountered in patients with atherothrombotic stroke. No specific findings by TEE were seen in patients with lacunar stroke. TEE is capable of detecting definite etiologies in cardioembolic stroke and associated cardiac pathologies in atherothrombotic stroke and lacunar stroke. These observations suggest that TEE is a useful tool to guide the physician for the treatment of ischemic stroke patients. PMID- 9201108 TI - Significance of downsloping ST-segment depression induced by low-level exercise in severe coronary artery disease. Assessment with myocardial ischemia and collateral perfusion. AB - Exercise-induced downsloping ST-segment depression is a common manifestation of severe myocardial ischemia. Although greater downsloping ST-segment depression is suspected to indicate more severe ischemia, its exact relationship to regional myocardial blood flow (RMBF) has not yet been clarified. We investigated the relationship between the magnitude of downsloping ST-segment depression and exercise-induced changes in RMBF and collateral perfusion. Nitrogen-13 ammonia positron emission tomography was performed in 6 healthy volunteers and 72 patients with angiographically proven coronary artery disease. The left ventricle was divided into 11 regions of interest, and RMBF in each region was measured at rest and during low-level supine bicycle exercise. Downsloping ST-segment depression of 0.1 mV or more at 80 milliseconds after the J point was accepted as significant. Low-level exercise induced downsloping depression of 0.1 to 0.2 mV in 10 patients (group D1) and downsloping depression of 0.2 mV or more in 8 patients (group D2). Multivessel disease was common in both group D1 (80% of patients) and group D2 (88% of patients). Collateral circulation was significantly more frequent in group D1 (90%) than in group D2 (13%, p < 0.01). Ischemic areas were larger and cardiac function was worse in group D2 than in group D1. The RMBF increased sufficiently in all regions (56 +/- 30%) with exercise in the healthy group. In group D1, RMBF was unchanged or decreased in ischemic areas (10 +/- 23%) but increased sufficiently in surrounding areas (50 +/- 32%). In group D2, RMBF was unchanged in ischemic areas (17 +/- 24%) and increased insufficiently in surrounding areas (41 +/- 21%). Therefore, exercise induced downsloping ST-segment depression of 0.1 to 0.2 mV may reflect an underlying change in blood flow in viable myocardium with collateral perfusion, and downsloping depression of 0.2 mV or more may reflect more severely impaired myocardium without collateral perfusion. PMID- 9201109 TI - Influence of exercise on QT dispersion in ischemic heart disease. AB - QT dispersion (QTd: maximum QT interval-minimum QT interval) is associated with severe cardiac arrhythmia and with abnormal ventricular repolarization. We investigated the influence of exercise on QTd in patients with ischemic heart disease. On standard 12-lead electrocardiograms, QTd was measured before and after treadmill exercise in 7 normal subjects, 17 patients with effort angina pectoris (and > or = 75% stenosis on coronary arteriography), and 33 patients with old myocardial infarction. Bazett's formula was used to obtain the corrected QTd (QTcd). The pre-exercise resting QTcd was 45.9 +/- 10.6, 44.3 +/- 15.2, and 74.8 +/- 28.1 msec in the respective groups, being significantly greater in the infarct group (p < 0.05). The QTcd at 5 min after exercise was respectively 49.3 +/- 9.0, 58.8 +/- 19.9, and 75.4 +/- 30.9 msec (p = 0.0347, infarct vs. controls). The difference in QTcd was significant for the angina group before and after physical exercise (p = 0.0003). There was a significant increase of QTcd after exercise in the angina group whether or not the patients were receiving beta-blockers. The infarct patients without beta-blocker therapy showed an increase of QTcd after exercise, while those receiving beta-blockers showed a decrease. The post-exercise difference between these subgroups was significant (p = 0.0351). CONCLUSIONS: QTcd was significantly increased by exercise in the angina group, possibly reflecting impaired repolarization due to ischemia. Inhibition of the increase in QTd by beta-blockers suggested a possible preventive effect on severe arrhythmias due to nonhomogeneous ventricular repolarization. PMID- 9201110 TI - Lack of association between angiotensin-converting enzyme gene polymorphism and coronary heart disease in a Chinese population. AB - Insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme (ACE) gene has been postulated as a risk factor for coronary heart disease. We conducted a case-control study of 271 Chinese, including 114 subjects with coronary artery disease (CAD), 42 with non-CAD and 115 apparently normal controls to examine the association of I/D polymorphism and CAD. The genotypes were identified by polymerase chain reaction and the plasma ACE activity was assayed by spectrophotometry. The allele and genotype frequencies were not different among the CAD, non-CAD and apparently normal groups (p = 0.42 and 0.63). Plasma ACE activity was not different among the three groups (p = 0.32). The D-allele and DD genotype were not more prevalent in subjects with low risk CAD (p = 0.07 and 0.16) and subjects with myocardial infarction (p = 0.79 and p = 0.35). No association was found between I/D polymorphism and severity of CAD (p = 0.42 and 0.70). In conclusion, the deletion polymorphism of the ACE gene may not be an independent risk factor in the development of CAD or myocardial infarction in this Chinese population. The unique or synergistic effect of other genes needs further study. PMID- 9201111 TI - Calcium antagonists and prevention of ventricular fibrillation induced by transient or persistent ischemia. AB - Experimental studies have shown the limitation by calcium antagonists of the propensity to fibrillation secondary to the occlusion of a large coronary artery. However, this capacity, studied in the acute phase of infarction, is less obvious and still under debate. Ischemia was therefore produced in anesthetized, open chest pigs by complete occlusion of the left anterior descending coronary artery according to two modes, either near its origin during brief but increasing periods (30, 60, 120, 180 s, etc) or half-way from this origin for a much longer time (60 min). The time course of vulnerability to fibrillation was monitored by ventricular fibrillation threshold (VFT), measured by trains of diastolic stimuli of 100 ms. Verapamil was administered in a 50 micrograms/kg dose followed by 2 micrograms/kg/min infusion. 1) In the case of brief proximal occlusions under pacing at a constant high rate (180 beats/min), verapamil slowed the decline of VFT from 6-8 mA to nearly 0 mA. VFT was 4.4 +/- 0.4 mA after 60 s ischemia, whereas it had already fallen to 1.8 +/- 0.3 mA (p < 0.001) in the absence of the drug. Accordingly, the onset of spontaneous fibrillation which depends on the decrease in VFT to about 0 mA was prolonged from 2-3 to 6-9 min. Bradycardia, concurrently produced by verapamil, is a factor which enhances these alterations. 2) In the case of a persistent midportion occlusion of the artery under sinus rate, fibrillations were similarly delayed by verapamil from 14-25 to 23-49 min after occlusion, but they were more numerous. VFT was lowered to critical values later, but also for a longer time. The period propitious to fibrillation was prolonged because the return of VFT to higher values reflecting hypoexcitability subsequent to the first cell injury was substantially delayed. Consequently, calcium antagonists should often prevent ventricular fibrillation when transient ischemia disappears before VFT falls to the vicinity of 0 mA. In contrast, a real benefit could not be expected from these drugs when ischemia is persistent since they then only delay fibrillations, the number of which is increased. PMID- 9201112 TI - Ventriculo-arterial coupling and the areas under the end-systolic pressure-volume relation. AB - Ventriculo-arterial coupling is expressed as the ratio Emax/eam (maximum ventricular elastance/arterial elastance). Different areas under the end-systolic pressure-volume relation (ESPVR) are expressed in terms of Emax/eam. The explicit inclusion of the active force of the myocardium in the mathematical formalism describing the pressure-volume relation (PVR) leads to new insight into the mechanics of left ventricular contraction. Applications to experimental data related to stroke work area SW under ESPVR are discussed and provide further evidence for the consistency of the mathematical formalism used. PMID- 9201113 TI - Effect of chronic digoxin on beta-adrenergic receptors in rabbits with heart failure. AB - This study investigated the effect of chronic digitalis glycoside use on beta adrenergic sympathetic activities in heart failure. Twenty-two Japanese white rabbits were anesthetized by intravenous injection of chloral hydrate. Aortic regurgitation (AR) was produced by perforating aortic valves in 14 rabbits. Digoxin was given for 1 week to 7 rabbits with AR (AR + Dig) and saline to 7 rabbits with AR (AR + C). Sham operation was performed in the remaining 8 rabbits (S). The left ventricular end-diastolic pressure was higher in AR + C than S (p < 0.05). It was lower in AR + Dig than AR + C (p < 0.05). Cardiac output was lower in AR + C than S (p < 0.05). There was no difference between AR + Dig and S. Both the left ventricular end-diastolic and end-systolic diameters were larger in AR + C (p < 0.05) than S, but they were similar between AR + Dig and S. Plasma norepinephrine level was lower in AR + Dig than AR + C. Myocardial beta adrenergic receptors number determined by radioligand binding assay using 30-800 pM 125I-iodocyanopindolol was lower in AR + C than S (28.8 +/- 7.9 vs. 69.9 +/- 12.3 fmol/mg protein, p < 0.05). It was higher in AR + Dig (39.9 +/- 9.8) than AR + C (p < 0.05). Myocardial norepinephrine content was lower in both AR + C (p < 0.05) and AR + Dig than S (p < 0.05). Thus, digitalis glycosides exert favorable effects on beta-adrenergic sympathetic activities in addition to the effects on hemodynamic variables in this animal model of heart failure. PMID- 9201114 TI - Serum angiotensin-converting enzyme and plasma atrial natriuretic peptide levels in hyperthyroid and hypothyroid rabbits. AB - BACKGROUND: It is known that serum angiotensin-converting enzyme (ACE), and plasma atrial natriuretic peptide (p-ANP) levels increase in hyperthyroidism. However, the precise mechanism of the effects of thyroid hormone on ANP release remains to be clarified. No study investigating serum ACE together with p-ANP levels has been performed in experimental hyperthyroid and hypothyroid rabbits. The present study was designed in order to provide additional evidence of increased ANP production and secretion in hyperthyroidism and to investigate the relationships between ANP, ACE and thyroid hormones. METHODS: Male New Zealand white rabbits (2.3-3.4 kg) were used throughout the study. Hyperthyroidism was induced by daily intraperitoneal administration of L-thyroxin (50 micrograms/100 g). Hypothyroidism was induced by daily intraperitoneal injection of propylthiouracil (2 mg/100 g body weight). Twelve days after the end of treatment, animals were sacrificed under anesthesia and blood samples were obtained from the aorta for serum ACE and thyroid hormone and p-ANP determinations. RESULTS: Serum ACE, plasma renin activity (PRA) and p-ANP were higher in hyperthyroid rabbits and lower in hypothyroid rabbits than in euthyroid rabbits. ANP concentration in atria was lower in hyperthyroid rabbits and higher in hypothyroid rabbits than in euthyroid rabbits. p-ANP, PRA and serum ACE levels were positively correlated with serum thyroxin levels. Inverse correlation was found between serum thyroxin and ANP concentration in atria (a-ANP), and between p-ANP and a-ANP. CONCLUSIONS: Our results indicate that not only p-ANP but also serum ACE activity was markedly increased in experimental hyperthyroid rabbits. It was thought that there were both direct and indirect effects of thyroxin on the release of ANP. PMID- 9201115 TI - Laminin alpha 1, alpha 2, alpha 4 and beta 1 chain mRNA expression in mouse embryonic, neonatal, and adult hearts. AB - We examined the relative expression of laminin alpha 1, alpha 2, alpha 4 and beta 1 chain genes in the heart of embryonic, neonatal and adult BALb/c mice. The reverse transcriptase-polymerase chain reaction was employed to determine the mRNA expression of these chains because of the relatively small amount of RNA extracted from rat embryonic hearts. Glyceraldehyde 3-phosphate dehydrogenase and beta-actin were used as internal controls. Among the hearts examined, a relatively high expression of laminin alpha 1 chain was observed in the embryonic hearts, while its expression was very weak in the neonatal and negligible in the adult hearts. Conversely, expression of laminin alpha 2 chain was virtually not observed in the embryonic hearts, but this chain was expressed weakly in the neonatal and substantially in the adult hearts. Similar to laminin alpha 1, laminin alpha 4 was expressed in the embryonic hearts, while its expression was relatively weak in the neonatal and adult hearts. Laminin beta 1 was expressed in the hearts of mice at all stages examined. These results demonstrate that the laminin chain gene expression changes in the different developmental stages of the hearts of BALb/c mice. PMID- 9201116 TI - Intraatrial rerouting in left isomerism and late complications. AB - We performed a total repair for a 1-year-old boy with left isomerism, a common atrium, incomplete atrioventricular septal defect and hemiazygos continuation. The operation performed was intraatrial rerouting with a baffle and mitral repair. The postoperative course was uneventful. However, two and a half years after the total repair, the patient revealed obstruction of the systemic atrium constructed by the baffle and the left ventricular outflow tract. Reoperation to relieve these obstructions was successful. PMID- 9201117 TI - Surgical treatment of acute myocardial ischaemia related to coronary angioplasty with special reference to use of perfusion balloon catheter and long-term outcome. AB - Twenty of 569 consecutive patients (3.5%) undergoing percutaneous transluminal coronary angioplasty required emergency coronary artery bypass grafting for acute closure of the dilated vessel. In seven patients an intracoronary autoperfusion balloon catheter was inserted to ensure antegrade blood flow across the injured zone of the coronary artery. The time needed for completion of the bypass grafts ranged from 100 to 399 minutes (mean 180 minutes). An average of 1.9 coronary artery bypasses was inserted. In total, 11 of the 20 patients (55%) developed new Q waves and had elevated CK-MB levels. However, the myocardial infarction rate was only 14% in those with a perfusion balloon catheter as against 77% in those without one. The insertion of a ball-out catheter permitted greater utilization of the internal mammary artery as a bypass graft. Angiographic follow-up was conducted after a mean of 28 months (19 patients). The patency rate of the bypass grafts placed in the emergency setting was relatively good (91%). Thallium tomography revealed a scar of variable size in all 17 patients studied and a reversible exercise perfusion defect requiring coronary reangioplasty in three patients. In conclusion, the insertion of a perfusion balloon catheter after abrupt coronary occlusion during coronary angioplasty solved the problems of acute myocardial ischemia and markedly lowered the definite myocardial infarction rate. This technique ensures favourable haemodynamic conditions for emergency myocardial revascularization. PMID- 9201118 TI - Effect of anticoagulants on the incidence of late pericardial tamponade following open heart surgery. The Hawaii experience. AB - We report on 17 cases of late pericardial tamponade (LPT, > 60 hours postoperatively) occurring post aortic and/or mitral valve replacement or coronary artery bypass graft surgery between 1979 and 1994. This includes one patient in whom LPT occurred twice. These cases were found from a search of 374 patients including those who were diagnosed with hemorrhagic complications secondary to open heart surgery, pericardial effusion and tamponade, those who underwent pericardiocentesis and a randomly picked group of patients. The mean age of the group was 57.8 years and included 11 males and 6 females. Due to the relatively small size of our sample (reflecting the infrequency of this complication) we force matched this tamponade group to look for any relationships that may exist between the incidence of LPT and anticoagulant therapy. No significant difference was found between the two groups with documented preoperative anticoagulant therapy (number of days; p > 0.2) or in relation to coagulation tests (prothrombin time, partial thromboplastin time and platelet counts; p > 0.2). In our case series, anticoagulant therapy did not appear to significantly affect the incidence of LPT. PMID- 9201119 TI - Primary repair of interrupted aortic arch and associated heart lesions in newborns. AB - Primary repair of interrupted aortic arch and associated heart lesions was performed in 13 patients aged from 1 to 85 days. The surgery was performed through the midline sternotomy approach in extracorporeal circulation and deep hypothermia. Hypothermic circulatory arrest at 14 to 19 degrees C was used for reconstruction of the aortic arch. In all patients it was possible to perform a direct anastomosis between the ascendent and descendent aorta. At the same time closure of the ventricular septal defect was performed in 11 patients, closure of the atrial septal defect in 4, correction of persistent truncus arteriosus in 3, resection of subaortic stenosis in 2, arterial switch repair of transposition of the great arteries in 1, correction of double outlet right ventricle in 1 and patch closure of aortico-pulmonary window in 1 patient. Three (23.1%) newborns died in the early postoperative period: two from sepsis and one from multiple organ failure. Ten patients (76.9%) were followed up for 1 to 29 months postoperatively. All of them are in very good condition with a nonrestrictive aortic anastomosis. Primary one-stage repair of interrupted aortic arch and associated heart lesions is preferred to the two-stage repair in all newborns with this critical congenital heart disease. PMID- 9201120 TI - Aneurysms of the sinus of Valsalva. AB - OBJECTIVE: To evaluate the properties of the coexistent cardiac anomalies associated with the aneurysm of sinus of Valsalva (ASV) and examine the long-term surgical results after operation. PATIENTS: From 1980 to 1994, nine patients (median age 22 years) underwent surgical correction of ASV. Aneurysms originated from the right (n = 5), noncoronary (n = 3) and left coronary sinus (n = 1) and entered into right ventricle (n = 5), right atrium (n = 3). In one patient, ASV originated from the left coronary sinus and unruptured. Coexistent cardiac lesions were aortic valve insufficiency (n = 4), ventricular septal defect (n = 5), patent foramen ovale (n = 1), right ventricular outflow tract obstruction (n = 1) and coronary artery anomaly (n = 2). All patients were symptomatic (sudden onset of symptoms in 3, gradual onset in 6). INTERVENTIONS: Ruptured ASVs were repaired by double approach in which both the involved chamber and the aortic root. Concomitant aortic surgery was performed in four patients (2 replacement, 2 valvuloplasty). VSDs were closed by patch in 4 and by direct suture in 1. RESULTS: The incidence of coexisting coronary artery anomaly was 22.2%. There was no hospital and late mortality. The mean follow-up period was 6.8 years (range 1 to 14 years). There were no reoperation for leaks of VSD, recurrence of aneurysm and aortic regurgitation. Eight patients were found to be in New York Heart Association class I, one patient in class II. CONCLUSION: The risk of the recurrent fistula or VSD is prevented by double approach technique, and also this approach reduces the incidence of late aortic insufficiency. Routinely preoperative coronary angiography must be performed for determine of coronary anomaly. PMID- 9201121 TI - Williams-Beuren syndrome. Long-term results of surgical treatments in six patients. AB - To settle long-term outcome after surgery for supravalvular aortic stenosis in the Williams-Beuren syndrome, we reviewed the records of 6 patients who had repair of the localized form (n = 5) or diffuse form (n = 1) at our Institution from 1965 to 1971. Four patients were females and 2 males, ages at operation ranged from 9 to 16 years (mean = 13 +/- 2.37 years). In all the patients was present the typical elfin facies with mental retardation and reduced I.Q. Preoperative omeral pressure was different between left and right arm (89 +/- 7/67 +/- 8 vs 105 +/- 8/77 +/- 4). Chest X-ray showed and enlargement of the cardia silhouette in all the patients. Cardiac catheterization, performed in all the patients, allowed diagnosis of supravalvular aortic stenosis and, in one case of subaortic stenosis associated. Intraoperatively a coronary tree enlargement was found in all cases with particular involvement of the right coronary in two patients. The mean diameter of the ascending aorta was 5.67 +/- 1.97 mm but the smallest (3 mm) was in the diffuse group. In group with localized stenosis the aortic root was enlarged with a teardrop patch in Dacron (n = 4) or a simple transverse suture after a longitudinal incision (n = 1). A pantaloon-shaped patch was necessary in the diffuse form case. There were no operative deaths and all the patients were discharged from the hospital after 2 weeks. A clinical follow up was possible in all the patients (10%) extended from 25 to 30 years (mean = 27.7 +/- 2.19 years); there were no late deaths and at presents time the mean age of the patient is 40 +/- 3 years. All patients were in functional class I or II. There was no significant difference between patients with a teardrop-shaped or pantaloon-shaped patch in terms of late gradient, survival, or aortic insufficiency studied by Echocardiography and color-Doppler. Of six patients two are living with parents or relatives but four are in a farm-college for disable people working and having some responsibility. We conclude that surgery for the correction of supravalvular aortic stenosis in Williams-Beuren syndrome is mandatory and both the procedures with patch techniques provide excellent long term results of gradients and aortic valve competence. Moreover the patients after the operation can have a normal activity with a satisfactory style and expectation of life. PMID- 9201122 TI - Surgical management of endocarditis-induced aorto-atrial and aorto-ventricular fistula: a new technique. AB - Early onset aortic prosthetic valve endocarditis with annular extension of the infectious process remains a highly lethal disease. Destruction of the annular tissue requires complex techniques for repair. The most challenging situation is associated with destruction and fistula formation at the left fibrous trigone. We report a technique using the anterior mitral leaflet to repair an aorto-atrial and aorto-ventricular fistula. This technique places autologous tissue from the anterior leaflet of the mitral valve against the infected area, and may decrease the risk of reinfection. PMID- 9201123 TI - Ductus diverticulum aneurysm associated with bicuspid aortic valve and dilatation of ascending aorta. AB - Saccular aortic aneurysm arising in the ductal region, known as ductus diverticulum aneurysm, is a rare anomaly. Due to potentially malignant evolution, recognition of radiological signs associated with this anomaly during the asymptomatic phase may be important in order to plan surgical intervention in time. We report an unusual case of ductus diverticulum aneurysm combined with bicuspid aortic valve, dilatation of ascending aorta, and coronary artery disease. PMID- 9201124 TI - Classification of the subclavian steal syndrome with transcranial Doppler. AB - Ultrasound has provided a highlight of the different types of subclavian steal. The authors report epidemiological and clinical data concerning 40,000 ultrasound examinations performed on epiaortic arteries and particularly the last 12,000 in which Doppler c.w., duplex scanner and transcranial Doppler were used. Various types of steal are described; five types of subclavian steal have been classified and patients stratified as being symptomatic and asymptomatic. The neurological symptoms are divided as follows: generalized cerebral ischemia, vertebro-basilar ischemia and hemispheric ischemia. Based on this clinical and haemodynamic outline, surgical therapy is indicated and type of surgery suggested. PMID- 9201125 TI - Operative results of thoracoabdominal repair for chronic type B aortic dissection. AB - From January 1991 to May 1994, we have operated on 15 cases of Type B aortic dissection. In 10 of these patients, thoracoabdominal repair was performed. According to Crawford's classification, 2 patients fell into Type I, 6 patients into Type II, and 2 patients into Type III. The aneurysms were exposed through a left thoracotomy extending into the retroperitoneum with the hemidiaphragm divided circumferentially. The operations were performed under femoro-femoral partial cardiopulmonary bypass. In 6 of these cases selective perfusion of the visceral branches was used. The celiac axis was reconstructed in 10 patients, superior mesenteric artery in 9, right renal artery in 7, left renal artery in 6. Abdominal vessels were reconstructed by the "inclusion" technique described by Crawford in 2 patients, by "beveling" the distal prosthetic end in 6 and by the "interposition" technique in 4 patients. Vessels arising from the false lumen were reconstructed by the "interposition" technique. To prevent paraplegia, the evoked spinal cord potentials by direct stimulation of the cord (ESPs-dsc) were monitored perioperatively and the aneurysms were repaired sequentially in segments. In all patients except 2 with Crawford type III aneurysms, spinal cord ischemia was detected by ESPs-dsc. In 7 of these patients, 2 to 8 pairs of intercostal/lumbar arteries (I/L aa.) that arose from the "responsible" aortic segment were reconstructed. Reconstruction techniques included the "inclusion" technique in 2 patients, the "beveling" technique in 1, the "interposition" technique in 1 and the "on lay grafting" technique in 3 patients. One hospital death occurred in a patient who had chronic renal insufficiency and liver cirrhosis preoperatively. Spinal cord injury occurred in 5 patients, including 4 paraparesis and 1 delayed-onset paraplegia. In 2 of these patients, responsible I/L aa., were not reconstructed correctly despite ESPs changes, and injury might have been prevented if reconstruction of the "responsible" arteries had been performed. Thoracoabdominal repair for chronic Type B aortic dissection could be performed safely with an acceptable mortality rate. Spinal cord injury remains an unsolved problem. PMID- 9201126 TI - Tourniquet occlusion technique for lower extremity artery reconstruction in war wound. AB - Reconstruction of blood vessels after war injuries is mandatory for life and limb salvage. In an effort to prevent prolonged major bleeding and make reconstruction quicker and technically more comfortable, thigh pneumatic tourniquet occlusion was performed pre- and intraoperatively in eleven of 53 wounded with injuries of the arteries of the lower extremities during 1991/92 war against Croatia at the Department of Surgery, Osijek Clinical Hospital. This method benefits life and limb salvage under war conditions. PMID- 9201127 TI - The successful management of a non-atherosclerotic ilio-femoro-popliteal aneurysm. Case report. AB - The successful management of an extensive non-atherosclerotic ilio-femoro popliteal aneurysm is described. This very rare aneurysm developed at the site of an old penetrating lower thigh wound caused by an infected arrow. To the best of our knowledge this is the first reported case in English literature. PMID- 9201128 TI - Aneurysmal arterial disease in a patient with Ehlers-Danlos syndrome. Case report and literature review. AB - Multiple aneurysmal lesions and dissections of the right femoral artery in a young man with type IV Ehlers-Danlos syndrome are presented. Type IV results in a high incidence of vascular lesions-extreme fragility of arteries is associated with multiple aneurysm formation and spontaneous rupture and dissection of arteries. Surgical management of patients with this disorder is hazardous and often unrewarding. The successful surgical treatment of an acutely thrombosed aneurysmal femoral artery by means of an interposition graft, is presented and brief guidelines for the treatment and prevention of vascular complications are discussed. PMID- 9201129 TI - Ruptured aortic pseudo-aneurysm: a rare presentation as aortocaval fistula. AB - Pseudo-aneurysms after abdominal aortic replacement are rare, occurring in less than 1% of operated patients. Usually asymptomatic, they may present clinically as a pulsatile mass or less commonly complicated with rupture. Aortocaval fistulas are rare, usually related to ruptured aortic aneurysms or trauma. A case of aortocaval fistula secondary to a ruptured pseudoaneurysm in a 81 year old woman is reported. After an infrarenal aortic aneurysm repair, the patient remained asymptomatic for 7 years, but abdominal pain and syncope developed and promoted further investigation. She was found to have a 6 cm by 6 cm retroaortocaval false aneurysm which had ruptured into the inferior vena cava. Computed tomography with intravenous contrast suggested the diagnosis and color Doppler ultrasound failed to confirm it. The etiology and management of this rare case are discussed. PMID- 9201130 TI - Persistent sciatic artery aneurysm with ruptured internal iliac artery aneurysm. AB - A case of aneurysm of persistent sciatic artery with ruptured aneurysm of the internal iliac artery is reported. A 62-year-old man was admitted to our hospital in emergency because of abdominal severe pain. Abdominal CT and angiography showed aneurysm of the infrarenal aortic aorta and left internal iliac artery and left persistent sciatic artery. The persistent sciatic artery aneurysm was coursed down the left thigh. Left superficial femoral artery was supplying blood to the popliteal artery as normal connection. Emergency operation was performed. The left internal iliac aneurysm was ruptured to a retroperitoneal space. The aneurysm was replaced by a Y-shaped dacron double velour graft from the infrarenal abdominal aorta to right common iliac and left external iliac arteries, which was followed by a ligation at the origin of left persistent sciatic aneurysm. The pulsation of the left lower extremity was good and additional bypass procedure to the popliteal artery was not performed. The post operative course of the patient was excellent and post-operative angiography showed normal anatomy of the left lower extremity. Optimal management of ruptured abdominal aneurysm thus requires not only an emergency operation but detailed observation of preoperative CT and angiography in order to scrutinize the associated vascular anomalies. PMID- 9201131 TI - Combined surgical and endovascular treatment of a traumatic pseudo-aneurysm of the brachiocephalic trunk with anatomical anomaly. AB - The authors report a case of combined surgical and endovascular treatment of a traumatic pseudo-aneurysm of the innominate artery in which the left common carotid artery originated from the brachiocephalic trunk. After a conventional surgical intervention with the implantation of the left common artery on the left subclavian artery, to correct the anatomic anomaly, a safe and effective endovascular stent-graft placement excluded the aneurysm. This new technique proposes a good chance for polytraumatized patients to receive a better prognosis and a much faster rehabilitation. PMID- 9201132 TI - A new device for prevention of postoperative haematoma in the surgery of varicose veins. AB - The objective of this article is the presentation of a new device, simple, easy to use, at low cost, for the prevention of postoperative haematoma following surgery of varicose veins of the lower limbs. It consists in a two-part device that functions as an elastic and pneumatic bandage, that wraps thigh and leg, with the knee articulation free and that is placed immediately before the stripping of the saphenous vein when all the surgical wounds are closed, except the supramalleolar one. While the head of the stripper is pulled, the device is inflated by air with a compression of 40-50 mmHg and the last surgical wound is sutured. Pneumatic compression is held for 24-36 hours, allowing the patients to walk and, in the meantime, to control the colour and the temperature of the foot. The advantages of this device are: easy use and low costs; compression on the area of the saphenous vein and of the main collaterals; uniform but moderate pressure on all the limb circumference. PMID- 9201133 TI - The radiological appearance of atherosclerotic popliteal artery aneurysms: the "dog-leg" sign. AB - The radiological appearance of popliteal aneurysms is described. Althought the lumen of the artery is often of normal diameter there is elongation of the vessel which frequently results in an acute "dog-leg". This sign has not been previously described. PMID- 9201134 TI - Extended intravascular shunting in an experimental model of vascular injury. AB - PURPOSE: To examine the extended patency (> 24 hrs) of heparin-bonded intravascular shunts in a porcine model of vascular injury. PROCEDURES: Adult swine underwent bilateral, common iliac artery resection (n = 5) or bilateral common iliac vein resection (n = 5) and vessel replacement with interposition, heparin-bonded shunts. Three control swine had vessel dissection only. Hematologic and coagulation profiles were measured at baseline and 24 hrs. Limb perfusion was assessed at 24 hrs by clinical exam and angiography. RESULTS: At 24 hrs, all limbs in both shunt groups were well perfused. All arterial shunts were angiographically patent. No distal emboli were detected. Nine of 10 venous shunts were patent, seven were lined with non-occluding thrombus. No alterations in hematologic or coagulation profiles were noted. CONCLUSIONS: Heparin-bonded shunts remained patent in arteries for 24 hours. Shunts placed in the venous system were prone to thrombus formation but most remained patent. PMID- 9201135 TI - Carcinoid tumour of the lung. AB - A retrospective analysis of 29 patients with carcinoid tumour of the lung treated between 1980 to 1995 is presented. There were 15 females and 14 males with a mean age of 57 years (range 28-72). Fourteen of the 29 carcinoids were peripheral and the remaining 15 were central. Preoperative histology was available in 17. Surgical resection was carried out in 27 patients, one patient was unfit whilst the other patient had multiple liver metastases at presentation. Surgical treatment offered were lobectomy (n = 19), pneumonectomy (n = 3), sleeve lobectomy (n = 3) and segmentectomy (n = 2). Twenty three patients were stage 1 tumours, 3 were stage II and one was stage III and 1 was stage IV. Postoperative histology confirmed typical carcinoids in 24 patients and the remaining 5 were atypical. There was one perioperative death from massive pulmonary embolism and there was no morbidity. Overall five year survival rate for patients with carcinoid was 96.4%. Five year survival for typical carcinoid and that of atypical carcinoid was 100% and 77.8% respectively. Typical carcinoids carry an excellent prognosis and should be offered conservative lung resection, whilst atypical carcinoids which behave aggressively should be treated by radical lung resections. PMID- 9201137 TI - Placental blood transplantation and autologous banking--caveat emptor. PMID- 9201136 TI - Imaging of bronchial carcinoid tumors associated to Cushing syndrome with 111In Octreoscan scintigraphy and immunoscintigraphy with anti-chromogranin monoclonal antibodies. Report of two cases. AB - Bronchial carcinoid tumors are neuroendocrine neoplasms capable of expressing somatostatin receptors and of secreting neuromediators such as ACTH and chromogranins. Radiologic appearance is usually non-specific and has to be distinguished from benign pulmonary nodules and other malignant diseases. Standard radiological techniques have limited accuracy in the evaluation of such lesions. Radioisotopic imaging techniques may increase the specificity of diagnostic assessment. The role of immunoscintigraphy with anti-chromogranin A and B monoclonal antibodies (MoAbs) and of 111In-Octreoscan scintigraphy is evaluated in two cases of bronchial carcinoid tumors associated to Cushing syndrome. PMID- 9201138 TI - Pediatric phase II cancer chemotherapy trials: a Pediatric Oncology Group study. AB - PURPOSE: This study reviewed the Pediatric Oncology Group experience with phase II clinical trials in children (< 21 years of age) with refractory tumors. PATIENTS AND METHODS: Patients registered in Pediatric Oncology Group phase II studies were evaluated. Patients had to be < 21 years of age with recurrent and refractory measurable disease. Tumor types and response rates were determined. Death on therapy from either drug toxicity, progressive disease, infection, or hemorrhage was measured. Tumor-specific, disease-free survival curves were calculated by Kaplan-Meier analysis. RESULTS: Between 1984 and 1994, 2,465 patient entries were made on 45 phase II trials. Malignancies registered included acute lymphocytic leukemia (ALL) (16.7%), acute myeloid leukemia (AML) (12.0%), osteogenic sarcoma (7.8%), neuroblastoma (7.2%), astrocytoma (7.2%), medulloblastoma (7.1%), glioma (6.7%), ependymoma (6.1%), and others (29.2%). The overall response rate was 19.6% (CR + PR) for children entered on phase II trials. Tumor-specific response rates ranged from 62.1% (23/37) for children with Hodgkin's disease to no responses (0/23) in patients with hepatoblastoma. When comparing single versus multiagent trials, a significantly better initial response rate was seen in the latter studies. However, 5-year survival was comparable. Progression-free survival for all tumor histologies were 12.9% and 9.2% at 2 and 5 years, respectively. Death on study was seen in 11.6% of the patients; however, only three deaths were directly related to drug toxicity. There were no significant gender differences in regards to response, progressive disease, or death on study. CONCLUSION: Phase II studies conducted in children offer a considerable likelihood of therapeutic benefit without exposing these patients to untoward toxicity. PMID- 9201139 TI - Correction of iron deficiency with an iron-fortified fluid whole cow's milk in children: results of a pilot study. AB - PURPOSE: This study assesses the efficacy of an iron-fortified (15 mg Fe, as stabilized ferrous sulfate (SFE-171), per liter) fluid whole cow's milk (IFFWCM) for the treatment of mild iron deficiency in children. Previous studies in healthy adult volunteers showed a mean 10.2 +/- 4.7% iron absorption. PATIENTS AND METHODS: Seventeen children (12 to 48 months old) with iron deficiency (serum iron (SI) < 60 micrograms/dl, transferrin saturation (TS) < 15%, serum ferritin (SF) < 15 ng/ml) were included in this study; 11 of them were anemic. As treatment, they received IFFWCM, instead of the customary whole cow's milk, for at least 4 months; medicinal iron was not administered. Hematocrit (Hct), hemoglobin (Hb), SI, TS, and SF were determined monthly. RESULTS: The Hb increased from 10.3 +/- 0.8 to 12.7 +/- 0.6 g/dl in the group with anemia (delta F-B: 2.4 +/- 1.0 g/dl) and from 12.6 +/- 0.7 to 13.5 +/- 0.3 g/dl in the group without anemia (delta F-B: 0.9 +/- 0.5 g/dl); the difference between both groups was significant (p < 0.01); the rate for Hct values showed a similar pattern. In the whole group, the SI increased to 84.8 +/- 37.4 micrograms/dl, with no difference between children with anemia and children without anemia; TS showed a similar pattern (delta F-B: 19.0 +/- 11.0%). The mean SF increased from 12.1 +/- 2.7 ng/ml to 27.9 +/- 25.4 ng/ml. Normal values for Hct, Hb, SI, and TS were reached by 100% of children; the rate for SF was 56.3%. Time required to reach normal Hct in the children with anemia was 59.4 +/- 33.0 days. Acceptance and tolerance were excellent; no treatment had to be discontinued. The group of patients with anemia was compared with an historical group composed of 55 children matched for age, basal Hct, and achieved Hct increase, treated with medicinal FS (4-6 mg/kg/day): time required to reach normal Hct was shorter in the FS-treated group (39.0 +/- 14.5 days) (p = 0.050). CONCLUSION: The use of IFFWCM alone could be an effective, relatively inexpensive, and well-tolerated treatment of iron deficiency in children. PMID- 9201140 TI - Safety and efficacy of low-dose intravenous immune globulin (IVIG) treatment for infants and children with immune thrombocytopenic purpura. Low-Dose IVIG Study Group. AB - PURPOSE: This report presents pooled data from two multicenter studies conducted to assess the efficacy, safety, and tolerance of lower-dose intravenous immune globulin (IVIG) regimens of 250 mg/kg/day, 400 mg/kg/day, and 500 mg/kg/day for 2 days, compared to an established higher-dose regimen of 1 g/kg/day for 2 days, in children with immune thrombocytopenic purpura (ITP). PATIENTS AND METHODS: A total of 24 children received IVIG (Gammar i.v.). In Study 1, 10 centers enrolled 12 children between 5 and 12 years old who received IVIG at either 400 mg/kg/day or 1 g/kg/day for 2 days. In Study 2, five centers enrolled 12 infants and children younger than 5 years old who received IVIG at 250 mg/kg/day or 500 mg/kg/day for 2 days. Both studies were prospective and randomized. RESULTS: IVIG treatment was effective (platelets increased at least 30,000/cu mm over baseline) in 94% (16 of 17) of the evaluable patients in the low-dosage group. Platelet increases occurred rapidly: by 48 hours, total platelet counts ranged from 32,000/cu mm to 256,000/cu mm, and peak platelet counts reached 38,000/cu mm to 551,000/cu mm. Adverse events (AEs) were most often mild, lasted less than 3 hours, and were usually those typically associated with immunoglobulin administration-headache, nausea, vomiting, and fever. There were two serious AEs an anaphylactoid reaction in one patient in the 400 mg/kg group and aseptic meningitis in one patient in the 1 g/kg high-dosage group. Both patients recovered without sequelae and were responders. Although the incidence of AEs varied by dosage groups, this difference was not significant. However, the incidence of AEs was affected by age. AEs were significantly lower in patients younger than 5 years of age. CONCLUSIONS: In this small, randomized trial, low dose IVIG in 2-day regimens of 250, 400, or 500 mg/kg/day rapidly reversed thrombocytopenia just as effectively as 1 g/kg/day in infants and young children with ITP. Lower-dosage regimens are safe and well-tolerated; the incidence of AEs is lower in children younger than 5 years of age. PMID- 9201142 TI - Early hospital discharge of children with fever and neutropenia: a prospective study. AB - PURPOSE: We report a prospective study on brief IV antibiotic therapy in selected children with cancer experiencing fever and neutropenia (FN) after chemotherapy. PATIENTS AND METHODS: All children with FN (T degree > or = 38 degrees C; ANC < 0.5 x 10(9)/L) were hospitalized for treatment with broad spectrum IV antibiotics. They were divided into three groups: group A (no infection), group B (clinically documented infection), and group C (bacteremia). Children in group A (and some children in group B) were discharged before recovery of neutropenia, if afebrile and in good condition. RESULTS: Eighty-eight consecutive episodes of FN occurred in 30 children. Children in group A (44 episodes; 50%) received IV antibiotics for a median of 3 days; on 25 occasions (57%), IV antibiotics were stopped before recovery of neutropenia. In children in group B (30 episodes; 34%), early discharge was allowed in eight cases of minor infections (27%); six received oral antibiotics. Two children (group A) were rehospitalized for recurrent FN but recovered without complications. CONCLUSION: In chemotherapy induced neutropenia, children hospitalized for fever but without documented infections and some children with minor infections can cautiously be discharged before evidence of bone marrow recovery if afebrile and in good general condition. PMID- 9201141 TI - Antiphospholipid antibodies and coagulation regulatory protein abnormalities in children with pulmonary emboli. AB - PURPOSE: To evaluate the demographics, presentation, family history, and laboratory findings in children with clinically recognized pulmonary emboli. METHODS: Data were collected about children with clinically recognized pulmonary emboli from 1987 to 1994 at two pediatric hematology referral centers. RESULTS: Sixteen children, mean age 11.8 years (standard deviation 4.69 years) including 11 boys were affected. Lower extremity thromboses were present in 7/14 children evaluated. Eight of the 16 children were apparently well before development of pulmonary emboli; seven were found to have antiphospholipid antibodies. None of the 15 children tested were antithrombin III deficient. One of 14 children tested was protein C deficient. Three of 13 children tested were protein S deficient or had a free protein S antigen at the fifth percentile. One of 10 children tested had an acquired dysfibrinogenemia. Two of nine children tested had the Factor V Leiden mutation. CONCLUSIONS: Our limited data suggest at least 70% of children with pulmonary emboli referred for hematology evaluation have antiphospholipid antibodies and coagulation regulatory protein abnormalities. PMID- 9201143 TI - Methods for tracing, contacting, and recruiting a cohort of survivors of childhood cancer. AB - PURPOSE: Due to the use of combined modalities of multiagent chemotherapy, radiation therapy, and surgery, many children with a diagnosis of cancer are now surviving into adulthood. This pilot study sought to determine the feasibility of establishing a cohort of childhood cancer survivors and then to develop methods to trace and contact eligible participants. MATERIALS AND METHODS: A retrospective cohort design was used. Four hundred and forty subjects who were treated for cancer at the University of Minnesota Hospital before the age of 21, between 1970 and 1986, had survived 5 years, and were alive at last contact were eligible. Tracing efforts were undertaken if the address was more than 2 years old or if a letter was returned by the post office. Contact procedures in this study were designed to determine whether participation rates differed according to the method of contact. RESULTS: In this cohort of 440 individuals, 11 had died and were not traced. Of the remaining 429 eligible individuals, 408 (95.1%) were successfully contacted. Successful tracing efforts differed by both current age and age at diagnosis. Once contacted, 370 (90.6%) agreed to participate in this study and returned a baseline health questionnaire. Each method of participation, and the combination of methods, showed similar percentages of participation. CONCLUSIONS: Results from this pilot study show that appropriate methods exist to establish a cohort of adults who have not been contacted since childhood. PMID- 9201144 TI - Late effects of therapy in survivors of Ewing's sarcoma family tumors. AB - PURPOSE: This late effects study was designed to determine if survivors of Ewing's sarcoma family tumors (ESFT) had adverse outcomes in employment, marital status, fertility, and functional status when compared to sibling controls. SUBJECTS AND METHODS: Eighty-nine survivors (case subjects) of ESFT treated at the National Cancer Institute between 1965 and 1992 and 97 sibling controls completed a questionnaire probing aspects of quality of life. The answers from case subjects were compared to pooled and matched sibling controls for all key variables. Odds ratios (OR) and p values from pooled analyses are presented. RESULTS: Although case subjects and controls did not differ in educational achievement, case subjects were less likely to be employed full-time (OR 0.4, p < 0.01), to be married (OR 0.2, p < 0.01), and to have children (OR 0.3, p < 0.01). Their most common treatment-related difficulties included permanent hair and skin changes (43%), lung problems (18%), neurologic problems (14%), visual difficulties (10%), second malignancy (7%), and amputation (5%). Functional status, measured by Karnofsky performance scale, was also adversely affected in case subjects. Case subjects did not differ from sibling controls in health care insurance status or in utilization of health services. CONCLUSIONS: Important aspects of life such as employment, marital status, fertility, and functional status are affected in survivors of ESFT. More studies are needed to better define the health status of adult survivors of pediatric cancer and the impact of cancer in adolescence on psychosocial development. PMID- 9201145 TI - Modulation of an acquired coagulation factor V inhibitor with intravenous immune globulin. AB - PURPOSE: We report that treatment of an immune mediated postoperative Factor V (FV) deficiency with intravenous immune globulin (IVIg) resulted in serological and clinical disappearance of the inhibitor. PATIENTS AND METHODS: A 9-year-old girl was exposed to bovine thrombin during cardiovascular surgery and subsequently developed severe, refractory hemorrhage caused by acquired FV deficiency (FV activity < 5%). Despite blood product transfusions, hemorrhage continued, and the patient was given IVIg, 400 mg/kg daily, for 9 day. RESULTS: Prolonged clotting times immediately trended toward normal, and the hemorrhage ceased by the fifth IVIg treatment day, concomitant with increasing plasma FV activity and disappearance of human FV inhibitor activity. The patient's plasma initially had a much higher inhibitor titer against bovine FV (122-215 Bethesda units) than against human FV (3-4 Bethesda units). Circulating antibodies (IgM and IgG) to bovine and human thrombin and FV were detected by enzyme-linked immunosorbent assay (ELISA). After completion of IVIg treatment, IgG antibodies to bovine FV and thrombin persisted, as did high-titer inhibition of bovine FV, whereas the subpopulation of IgG and IgM antibodies reactive with human FV were undetectable. CONCLUSIONS: The inhibitor likely developed from a heterogenetic immune response to bovine FV contaminating the topical thrombin preparation used during surgery. To our knowledge, this is the first demonstration of immunological clearance of an acquired FV antibody associated with the use of IVIg. The data suggest an antiidiotypic mechanism of IVIg in modulating clearance of antihuman FV antibodies. PMID- 9201146 TI - Treatment of childhood lymphangiomas with interferon-alpha. AB - PURPOSE: Nonsurgical treatment of lymphangiomas has shown limited efficacy and often carries unacceptable toxicities, demonstrating the need for a more effective, less toxic therapy. PATIENTS AND METHODS: We describe two patients with lymphangiomatosis treated for 12 to 40 months with recombinant interferon alpha. RESULTS: Both patients demonstrated stabilization or marked improvement of disease, based on clinical and radiologic findings, with minimal toxicity. CONCLUSIONS: The favorable responses to interferon-alpha therapy in these two cases suggest that this is an effective and well-tolerated treatment for lymphangiomas in children. PMID- 9201147 TI - Successful treatment of infantile hemangiomas with interferon-alpha-2b. AB - PURPOSE: Hemangiomas are benign tumors occurring in 10% of infants. A small percentage are complicated by blockage of vital structures, consumptive coagulopathy, or heart failure, resulting in a mortality of -20% of patients with complications. Here, we describe four infants with complicated hemangiomas responding to interferon-alpha-2b therapy. PATIENTS AND METHODS: Four children with hemangiomas were treated with interferon-alpha-2b for complicating heart failure (1), visual impairment (2), or coagulopathy (1). Patients received interferon-alpha-2b alone or in conjunction with corticosteroid therapy over 2 to 9 months. Imaging studies and urinary basic fibroblast growth factor (bFGF) levels were used to monitor treatment response. RESULTS: Three of four patients demonstrated involution of the hemangiomas with improvement in their coagulopathy or visual impairment. The fourth patient expired due to cardiac complications despite radiologic evidence of hemangioma involution. Side effects associated with interferon-alpha-2b treatment included elevated transaminases (2) and leukocytosis (2), which resolved upon completion of therapy. One patient developed mild gross motor delay (1), which improved after cessation of therapy. Decreased urinary bFGF levels correlated with hemangioma involution. CONCLUSION: Interferon-alpha-2b therapy is an effective, well-tolerated treatment for complicated hemangiomas. Measurement of urinary bFGF levels may provide an objective method for monitoring treatment response. PMID- 9201148 TI - Oral treatment in selective vitamin B12 malabsorption. AB - PURPOSE: The efficacy of oral treatment with megadose vitamin B12 in a patient with selective vitamin B12 malabsorption is studied. PATIENTS AND METHODS: An 8 year-old boy with megaloblastic anemia due to selective vitamin B12 malabsorption is presented. His history was significant for anemia of 4 years duration, requiring transfusion on two occasions. On admission, the Hb was 7.9 g/dL, WBC 6 x 10(9)/L, mean corpuscular volume 124 fl, red cell distribution width 16.8%, platelets 156 x 10(9)/L, reticulocyte 0.04%, and the serum vitamin B12 level 87 pmol/L. There was proteinuria. Replacement treatment with oral B12 1,000 micrograms/daily was instituted. RESULTS: Reticulocytosis was observed on the third day of treatment, which was followed by a gradual increase in Hb level to 12 g/dL in 3 weeks. A Schilling test performed after a 5-day interruption of therapy was compatible with malabsorption. CONCLUSIONS: Our study suggests that the oral route is as effective as the parenteral route when vitamin B12 is given at a dose larger than that of parenteral therapy. PMID- 9201149 TI - Preoperative chemotherapy for congenital hemangiopericytoma and a review of the literature. AB - PURPOSE: Malignant hemangiopericytoma is an uncommon tumor in the pediatric age group. A case of congenital hemangiopericytoma is presented with a review of the literature. PATIENTS AND METHODS: A 2-month-old boy presented at birth with an enlarging posterior neck mass. Subsequent histopathologic studies showed findings consistent with hemangiopericytoma. Imaging studies demonstrated an extensive, infiltrative, vascular lesion not readily amenable to surgical resection. A pulmonary metastatic lesion was present. RESULTS: Combined treatment using vincristine, doxorubicin, and cyclophosphamide given preoperatively resulted in a significant decrease in size of the primary tumor, allowing for resection without complication and resolution of the metastatic lesion. CONCLUSION: Preoperative chemotherapy may have a significant role in the management of infants with malignant hemangiopericytoma. PMID- 9201150 TI - Multifocal osteosarcoma in a patient with Fanconi anemia. AB - PURPOSE: The purpose of this report is to present a case of multifocal osteosarcoma in a patient with Fanconi anemia. PATIENTS AND METHODS: A nine year old girl with known Fanconi anemia presented with a pathologic femur fracture. Imaging studies confirmed soft tissue and bony involvement in the femur, ipsilateral tibia, and possibly contralateral femur. RESULTS: Biopsy and subsequent amputation confirmed the presence of classic osteosarcoma in the involved femur and ipsilateral tibia. The patient died seven months later of pulmonary failure secondary to metastases. CONCLUSION: Fanconi anemia is known to be associated with malignancies such as leukemias. We report a new association of Fanconi anemia with osteosarcoma. PMID- 9201151 TI - Cardiac rhythm abnormalities during intravenous immunoglobulin G infusion for treatment of thrombocytopenia. AB - PURPOSE: Several side effects of intravenous immunoglobulin G (IVIG) therapy are known, but it has never been reported to be associated with cardiac rhythm abnormalities other than sinus tachycardia. PATIENTS AND METHODS: We describe the development of cardiac dysrhythmias during intravenous immunoglobulin G infusion in two children with thrombocytopenia. One of the patients had a history of supraventricular tachycardia, and the other had evidence suggestive of preexisting long QT syndrome. CONCLUSION: Cardiac rhythm abnormalities may be exacerbated in individuals with preexisting cardiac problems by IVIG infusion, and such patients should be monitored closely during IVIG administration. PMID- 9201152 TI - Complications of the nevoid basal cell carcinoma syndrome: a case report. AB - PURPOSE: We report that patients with nevoid basal cell carcinoma syndrome (Gorlin syndrome) are at risk for developing neoplasms, especially basal cell carcinomas and rarely medulloblastoma. METHODS: A case report is presented of a 5 year-old child with medulloblastoma and multiple basal cell carcinomas who was diagnosed with nevoid basal cell carcinoma syndrome. Genetic analyses were performed on tumor DNA from the patient's medulloblastoma and basal cell carcinoma as well as germline DNA from the patient and unaffected family members. RESULTS: After radiation therapy for medulloblastoma, the patient developed thousands of additional basal cell carcinomas. Analysis of tumor DNA revealed the characteristic defect of nevoid basal cell carcinoma syndrome, loss of heterozygosity at 9q22. Photodynamic therapy was successfully used to control the majority of her cutaneous tumors. CONCLUSION: DNA analysis confirmed the presence of the distinctive genetic lesion of nevoid basal cell carcinoma syndrome in both medulloblastoma and basal cell carcinoma. Omitting or limiting radiation therapy for children with nevoid basal cell carcinoma syndrome and medulloblastoma should be considered. PMID- 9201153 TI - Wilms tumor associated with polycythemia: case report and review of the literature. AB - PURPOSE AND METHODS: A case of polycythemia with a normal serum erythropoietin is described, which led to the diagnosis of Wilms tumor. The clinical features of the reported cases of Wilms tumor associated with polycythemia are reviewed. RESULTS: An asymptomatic 6-year-old boy with polycythemia, a normal serum erythropoietin, and no evidence of erythroid colony forming activity in his serum was found to have a Wilms tumor. After resection and chemotherapy, he has had no recurrence of either the polycythemia or the Wilms tumor. There have now been 10 cases of Wilms tumor reported: 7 patients were more than 16 years of age, 8 were boys, and 9 were clinical stage I with a favorable histology. CONCLUSIONS: Polycythemia is a rare manifestation of Wilms tumor that can occur in the absence of an elevated serum erythropoietin and has an association with male gender, older patient age, and low clinical stage. Children with unexplained polycythemia should be investigated for Wilms tumor, even if the serum erythropoietin level is normal. PMID- 9201154 TI - Skin metastases of osteogenic sarcoma: a case report with review of the literature. PMID- 9201155 TI - Intrathecal leukovorin after intrathecal methotrexate overdose. PMID- 9201156 TI - "Single vision and Newton's sleep". PMID- 9201157 TI - Is coronary angiography necessary for vascular surgery patients who have positive results of dipyridamole thallium scans? AB - PURPOSE: Because dipyridamole thallium (DT) scanning is a useful predictor of perioperative cardiac events, a positive results of a DT scan is frequently the basis for performing more invasive cardiac evaluation and for consideration for performing coronary revascularization procedures before performing peripheral vascular surgery. The rationale for this approach has been that the treatment of anatomically significant coronary artery disease would lower the risk of performing a subsequent vascular operation. However, the benefit of performing aggressive diagnostic and therapeutic cardiac procedures in such patients remains unproved. To examine this issue, data from patients who underwent coronary angiography because of thallium redistribution were compared with data from matched control subjects who underwent peripheral vascular operations without further cardiac evaluation. METHODS: The medical records of 70 consecutive patients who underwent coronary angiography because of the presence of two or more segments of redistribution on DT scan were reviewed and compared with 70 other patients matched with respect to age, gender, peripheral vascular operation, and number of segments of redistribution on DT scan who did not undergo additional cardiac evaluation. RESULTS: DT scans were performed on 934 preoperative peripheral vascular surgery patients to help in the assessment of operative risk. Ischemic responses, defined as two or more segments of redistribution, were observed in 297. Of these, 70 underwent cardiac catheterization and 25 underwent coronary revascularization procedures. Adverse outcomes affected 46% of the coronary angiography group and 44% of the control group (p = NS). Patients who underwent coronary angiography and were considered for myocardial revascularization had fewer cardiac events with a subsequent vascular operation than did the control subjects. However, any possible benefit from invasive cardiac evaluation was offset by the three deaths and two myocardial infarctions (MIs) that complicated the cardiac evaluation. There was no significant difference between the angiography group and the matched control subjects with respect to perioperative nonfatal MI (13% vs 9%), fatal MI (4% vs 3%), late nonfatal MI (16% vs 19%), or late cardiac death (10% vs 13%). In long term follow-up, MIs occurred later in patients who underwent coronary angiography than the control subjects (p = 0.049), but this difference was not associated with an improvement in the overall survival rate. CONCLUSIONS: The risks of extended cardiac evaluation and treatment did not produce any improvement in either the perioperative or the long-term survival rate. For most vascular surgery patients who have a positive result of a DT scan, coronary angiography does not provide any additional useful information. PMID- 9201158 TI - Intraoperative autologous transfusion during elective infrarenal aortic reconstruction: a decision analysis model. AB - PURPOSE: The use of intraoperative autologous transfusion devices expanded during the last decade as a result of the increased awareness of transfusion-associated complications. This study was designed to determine whether routine use of an intraoperative autologous transfusion device (Haemonetics Cell Saver [CS]) during elective infrarenal aortic reconstructions is cost-effective ($50,000/QALYs threshold). METHODS: A decision analysis tree was constructed to model all of the complications that are associated with red blood cell replacement during aortic reconstructions for both abdominal aortic aneurysm (AAA) and aortoiliac occlusive disease (AIOD). It was assumed that a unit of CS return (CSR; 250 ml/unit) equaled a unit of packed red blood cells (PRBCs) and that all CS transfusions were necessary. Transfusion requirements (AAA:PRBC = 2.8 +/- 3.2 units, CSB = 3.7 +/- 3.2 units; AIOD:PRBC = 3.1 +/- 3.0 units, CSR = 2.1 +/- 1.7 units) were determined from retrospective review of all elective aortic reconstructions (AAA, N = 63; AIOD, N = 75) from Jan. 1991 to June 1995 in which the CS was used (82.1% of all reconstructions). Risk of allogenic transfusion-related complications (transfusion reaction, hepatitis B, hepatitis C, human immunodeficiency virus, human T-cell lymphotropic virus types I and II) and their associated treatment costs (expressed in dollars and quality-adjusted life years (QALYs) were obtained from the medical literature, institutional audit, and a consensus of physicians. RESULTS: Routine use of the CS during elective infrarenal aortic reconstructions was not cost-effective in our practice. Use during reconstructions for AAA repairs cost $263.75 but added only 0.00218 QALYs, for a rate of $120,794/QALY. Use during reconstructions for AIOD was even more costly at $356.68 and provided even less benefit at 0.00062 QALYs, for a rate of $578,275/QALY. The sensitivity analyses determined that the routine use of the CS would be cost-effective in our practice only for AAA repairs if the incidence of hepatitis C were tenfold greater than the baseline assumption. The model determined that CS was cost effective if the CSR exceed 5 units during reconstructions for AAA and 6 units during reconstructions for AIOD. CONCLUSIONS: The routine use of the CS during elective infrarenal aortic reconstructions is not cost-effective. The use of the device should be reserved for a select group of aortic reconstructions, including those in which cost-effective salvage volumes are anticipated. Alternatively, the CS should be used as a reservoir and activated as a salvage device if significant bleeding is encountered. PMID- 9201159 TI - Subfascial endoscopic perforator ligation: an analysis of early clinical outcomes and cost. AB - PURPOSE: Early results of subfascial endoscopic perforator surgery (SEPS) were examined. Data on ulcer healing, complications, and costs are presented. METHODS: Data were prospectively collected for all patients who underwent SEPS at our institution. A concurrent control group was not available because primary open perforator ligation is no longer performed at our hospital. Preoperative assessment included duplex scanning (valve closure times and perforator mapping), plethysmography, and phlebography. Completeness of therapy was assessed with postoperative duplex mapping of perforating veins. Clinical status was monitored after surgery, and actual costs, including equipment, personnel, and facilities management, are reported. RESULTS: Eighteen procedures were performed in 15 patients (mean age, 52 years; range, 42 to 65 years). Two patients underwent bilateral SEPS, and one patient underwent a second procedure on the same leg. Active ulceration (class 6) was present in 14 of 18 limbs (78%), recently healed ulcers (class 5) in two of 18 (11%), and lipodermatosclerosis with edema (class 4) in two. Deep venous insufficiency was present in 14 of 18 (78%). The number of perforating veins ligated per leg ranged from 0 to 12 (mean, 4.3). Follow-up ranged from 3 to 64 weeks (mean, 22 weeks). Complete ulcer healing occurred in eight of 14 limbs (57%) at a mean of 14 weeks. Reduction in ulcer size was noted in four of 14 (29%), and two limbs were not improved. There were no new ulcers. Residual perforating veins were noted in four of 18 limbs. None of the limbs with residual perforating veins had complete healing of ulceration. Operating room costs were higher than those associated with limited-incision open perforator ligation ($2570 vs $1883). CONCLUSION: These preliminary data suggest that when used as part of a treatment plan to correct deep and superficial venous insufficiency SEPS results in a high rate of wound healing, with no recurrent ulceration in this series. Increased operating room costs associated with longer operations and greater disposable expenses will likely be overcome by shortened length of stay and diminished wound complications. These findings emphasize the importance of ligating all incompetent perforating veins, as ulcer healing was never achieved when residual perforating veins were found at follow-up. PMID- 9201160 TI - Durability of early prosthetic dialysis graft cannulation: results of a prospective, nonrandomized clinical trial. AB - PURPOSE: Initiation of hemodialysis frequently requires temporary central venous catheterization, which leads to subsequent venous stenosis in 50% of patients. These lesions severely limit upper extremity dialysis fistula creation. The present study was undertaken to determine whether early cannulation (EC) allowed omission of temporary venous catheterization without affecting perioperative morbidity and long-term graft patency. METHODS: Seventy-nine prosthetic grafts for hemodialysis were placed in 76 patients over a 40-month period. Patients who required hemodialysis between 24 and 72 hours after surgery were assigned to EC. The remaining grafts underwent late cannulation (LC) after postoperative day 14. All grafts were constructed with a 6 mm stretch-expanded polytetrafluoroethylene conduit in the brachial artery-to-axillary vein position. Statistical analysis of cumulative primary patency estimates and patient survival data were determined by Kaplan-Meier analysis and log-rank test, patient variables were compared using chi 2 and Fisher's exact test, and multivariate analysis was performed using Cox's proportional hazard model. RESULTS: Forty-eight patients underwent EC and 31 underwent LC. There were no significant differences regarding age (mean, 61.5 years), history of diabetes, congestive heart failure, hematocrit level (mean, 30%), or presence of peripheral vascular disease. Thrombosis occurred before cannulation in one of 48 ECs (2.0%) and one of 31 LCs (3.2%). There were no episodes of cannulation hemorrhage or wound infection in either group. Cumulative primary patency estimates for EC were 0.89, 0.82, and 0.70 at 3, 6, and 12 months, respectively. These were not significantly different from the LC estimates of 0.86, 0.78, and 0.74 at 3, 6, and 12 months, respectively. Overall, patients who had a history of peripheral vascular disease had a significantly decreased 12-month patency rate (60% vs 74%; p = 0.05). Central venous catheters were omitted in 47 of 48 EC patients. CONCLUSION: EC of prosthetic dialysis grafts does not increase perioperative morbidity rates or decrease 12-month cumulative primary patency rates. PMID- 9201161 TI - Surgical complication outcome (SCOUT) score: a new method to evaluate quality of care in vascular surgery. AB - PURPOSE: Surgical outcome data are generally reported as raw morbidity and mortality rates, which do not necessarily reflect quality of surgical care. The Society for Vascular Surgery has led this area with recommendations by the Ad Hoc Committee on Reporting Standards to establish standardized methods of outcome assessment in vascular surgery. The purpose of this study was to evaluate a new method for evaluating the overall quality of surgical care, which includes surgeon, nursing, and hospital system performance. The goal of the method is to identify problem areas in surgical practice that can be targeted for focused improvement to improve outcome. METHODS: A database of more than 9000 general and vascular surgical cases was compiled over a 3-year period. Every postoperative complication was tabulated prospectively by a surgical nurse on a daily basis. Fifty clinically significant complication types specific for vascular surgery patients were identified from a list of 151 postoperative events by a panel of vascular surgeons and were grouped into nine broad categories (vascular, cardiac, pulmonary, etc.). These complications reflect the entire continuum of postoperative care, including surgeon, nursing, and hospital system performance. Each complication type was further stratified into four grades (mild, moderate, severe, death) and assigned a SCOUT severity score from 0 to 100 (0, no complication; 100, death) by the panel of surgeons. For case of data collection and monitoring of outcome, a software program was developed to run on a laptop computer and includes medical history, risk factors, pertinent laboratory data, and the preassigned SCOUT severity scores for measuring outcome. In this study, 170 major vascular procedures performed over the previous 12-month period were prospectively evaluated usig the SCOUT method in an attempt to more easily identify problem areas of practice. In-hospital morbidity and 30-day mortality results were examined. RESULTS: One hundred sixteen postoperative complications were identified in the 170 patients, with an overall morbidity rate of 51% and a 30-day mortality rate of 1.8%. Fifty-three percent of the complications were "mild" and required minimal intervention or observation only. Abdominal aortic aneurysm repair was associated with the highest morbidity rate (mean SCOUT score, 384.35), whereas distal extremity bypass grafting had the lowest morbidity rate (mean SCOUT score, 114.4). However, subgroup analysis demonstrated that cardiac events accounted for 52% of the morbidity associated with distal extremity bypass but only 34.7% of the morbidity associated with abdominal aortic aneurysm repair (p < 0.05). CONCLUSIONS: The SCOUT score is a new technical quality of care measure that can objectively quantify surgeon and other hospital system-related performance. The SCOUT score allows the surgeon to identify problem areas that can then be targeted for improvement to positively affect outcome. PMID- 9201162 TI - Hemodynamic impact of vein graft stenoses and their prediction in the vascular laboratory. AB - PURPOSE: We undertook this prospective evaluation to define the direct hemodynamic impact of vein graft stenoses and to correlate intraoperative hemodynamic findings with the preoperative duplex scan. METHODS: Twelve consecutive isolated vein graft stenoses were identified in the vascular laboratory during our routine duplex scanning surveillance protocol over 10 months. Peak systolic flow velocity ratios (PSFVRs; velocity within stenosis/velocity proximal to the stenosis) at the stenoses ranged from 2.7 to 10 (mean, 5.5), and ankle-brachial indexes ranged from 0.47 to 0.94 (mean, 0.68) Preoperative arteriograms were obtained and confirmed the isolated stenoses, which radiographically ranged from 20% to 83% diameter reduction (mean, 64%). At the time of surgery the stenotic graft segment was isolated, and simultaneous pressure measurements proximal and distal to the graft stenosis were measured, along with ultrasound transit-time blood flow measurements. Pressure and flow wave forms were recorded for 10 seconds at 200 Hz and were stored on a personal computer-based digital acquisition system. The graft stenoses were then repaired with either a vein patch or short interposition graft, and the hemodynamic measurements were repeated. Fourier transformation of the pressure and flow curves was performed, and the resistance and longitudinal impedance were calculated for each graft segment. RESULTS: Before repair, mean pressure gradients across the stenotic graft segments (delta P) ranged from 1.0 to 74.6 mm Hg (mean, 20.4 mm Hg), and vein graft flow ranged from 4.9 to 140 ml/min (mean, 45.2 ml/min). After repair of the stenotic segments, the mean pressure gradient was decreased to a mean of 1.3 mm Hg, and vein graft flow increased to a mean of 104.8 ml/min. The PSFVR recorded in the vascular laboratory correlated significantly with delta P (r = 0.71; p = 0.01) and allowed prediction of delta P as: delta P = 7.4 (PSFVR) - 19.8. PSFVR also correlated with measured resistance across the stenosis (r = 0.79; p = 0.004). Conversely, the angiographic measurement of stenosis did not correlate significantly with these parameters. The angiographic measurement of stenosis showed a moderate correlation with the PSFVR (r = 0.58; p = 0.046). CONCLUSIONS: The PSFVR, as measured in the laboratory, is an accurate and useful indicator of the hemodynamic impact of vein graft stenosis. Revision of stenotic vein graft segments resulted in a significant improvement in graft hemodynamics. PMID- 9201163 TI - Thrombolysis of occluded infrainguinal vein grafts: predictors of outcome. AB - PURPOSE: The purpose of this study was to identify factors that influence graft patency and limb salvage rates after thrombolysis of occluded infrainguinal vein grafts. METHODS: The records of patients who underwent percutaneous catheter directed thrombolysis of occluded infrainguinal vein bypass grafts at our institution between 1985 and 1995 were reviewed. Life table analysis was used to determine survival and patency differences. Univariate and multivariate analyses were used to identify the patient-specific factors that affected outcomes. RESULTS: Forty-four patients with 44 thrombosed infrainguinal vein grafts underwent thrombolysis with urokinase. The thrombolysis-related mortality rate was 2%, and nonfatal complications occurred in 16%. Thrombolysis was unable to restore graft patency in 25% of grafts (11 of 44). Of the remaining 33 successfully lysed grafts, 88% required adjunctive surgery or percutaneous transluminal angioplasty after thrombolysis. Overall, the primary graft patency rate was 25% at 1 year and 19% at 2 years after thrombolysis. Considering only successfully lysed grafts, the primary patency rate improved to 34% at 1 year and 25% at 2 years. Multivariate analysis revealed that the graft patency rate was substantially better in patients without diabetes and in vein grafts that had been in place for longer than 12 months (p < 0.01). The limb salvage rate was significantly improved by successful thrombolysis (63% at 2 years vs 31% if lysis failed; p < 0.01). The patient survival rate was high-89% 2 years after thrombolysis. CONCLUSIONS: Even with adjunctive therapy, vein graft thrombolysis is unlikely to yield durable patency overall. However, successful thrombolysis improves limb salvage rates and may be beneficial in patients without diabetes who have mature vein grafts but who do not have options for other autogenous revascularization procedures. PMID- 9201164 TI - Validation of a new and specific intraoperative measurement of vein graft resistance. AB - PURPOSE: Clinical studies have revealed that the most important predictor of successful bypass grafting is the origin and quality of the bypass conduit. Attempts at intraoperative evaluation of the hemodynamic properties of the conduit, including assessment of blood flow (Q), pressure gradients (delta P), and resistance (R), have not been useful. This is because each of these parameters measures the characteristics of the graft plus the outflow bed. To date, no specific measurement of the resistive properties of the conduit only is available. The purpose of this investigation was to evaluate longitudinal impedance (ZL) as a measure of conduit-specific resistance and to evaluate its potential in predicting the outcome of infrainguinal vascular reconstructions. METHODS: ZL was measured during surgery in 73 infrainguinal autologous vein reconstructions performed in 68 patients in two separate institutions over a 21 month period. Vein graft ultrasonic transit time Q and delta P (from proximal to distal anastomosis) were measured at baseline and after maximal peripheral vasodilatation with an intraarterial injection of papaverine 30 mg. Waveforms were recorded for 10 seconds at 200 Hz using a digital acquisition system. R was calculated as proximal mean pressure divided by mean blood flow (Q). After Fourier transformation, ZL was calculated as delta P/Q at each harmonic and total ZL (integral of ZL) was defined as the integral of moduli from 0 to 4 Hz. RESULTS: All hemodynamic variables were significantly affected by papaverine vasodilatation (delta P, 3.9 +/- 0.5 vs 6.3 +/- 0.8 mm Hg; Q, 78.2 +/- 7.0 vs 126 +/- 11 ml/min; R, 134 +/- 17 vs 72.7 +/- 6.2 x 10(3) dyne.sec.cm-5; p < 0.0001), except integral of ZL, which remained constant (31.1 +/- 2.8 vs 30.8 +/- 2.8 x 10(3) dyne.cm-5; p = NS). After follow-up of 1 week to 17 months (median, 5 months), the 1-year primary, primary-assisted, and secondary patency rates were 72% +/- 7%, 77% +/- 6%, and 81% +/- 6%, respectively. Using Cox analysis, primary patency was significantly associated with decreased integral of ZL (p = 0.0001), but not with baseline or papaverine-stimulated delta P, Q, delta P/Q, or R integral of ZL > 47 x 10(3) dyne.cm-5 predicted primary failure with 90% positive and negative predictive value. CONCLUSIONS: Intraoperative measurement of integral of ZL in infrainguinal vein grafts is independent of outflow conditions (that is, does not change with papaverine), and hence describes the resistive properties of the conduit only. In addition, these preliminary data suggest that integral of ZL is predictive of short-term primary patency. integral of ZL is the first available hemodynamic measurement that is conduit-specific and may therefore be a better predictor of graft patency than currently available methods. PMID- 9201165 TI - Photodynamic therapy inhibits transforming growth factor beta activity associated with vascular smooth muscle cell injury. AB - PURPOSE: The multifunctional cytokine, transforming growth factor beta 1 (TGF beta), plays an important role in the development of injury-associated intimal hyperplasia (IH). Strategies to suppress local TGF-beta activity may have a clinical potential to prevent restenosis caused by IH. Photodynamic therapy (PDT) involves the local generation of cytotoxic free radicals by light activation of photosensitizer dyes and has been shown to inhibit experimental IH. This study investigated whether PDT-generated free radicals can affect TGF-beta activity in a biologic system using vascular smooth muscle cells (SMCs). METHODS: The release and activation of TGF-beta by injured SMCs in culture was compared between mechanical injury and PDT. Mechanical injury was induced with a rubber policeman, and PDT was performed with the photosensitizer chloroaluminum sulfonated phthalocyanine (5 micrograms/ml) and 675 nm laser light at subtherapeutic 10 J/cm2 and the in vivo therapeutic dose of 100 J/cm2. Cell viability was assessed by the tetrazolium salt conversion assay, and active and total (active + latent) TGF-beta was determined by enzyme-linked immunosorbent assay in the conditioned media of SMCs 24 hours after treatment. Functional TGF-beta activity was assessed by inhibition of endothelial cell mitogenesis. RESULTS: Both forms of injury severely reduced (p < 0.0005) SMC viability to less than 15%. In untreated SMC conditioned media, only 14.5% of the total TGF-beta was active (27.7 +/- 8.7 pg per 1 x 10(5) cells). However, after mechanical injury and PDT with 10 J/cm2, there was a significant increase (p < 0.02) in active TGF-beta (60.1 +/- 10.1 pg and 48.6 +/- 21.0 pg, respectively), despite a total reduction of approximately 50%. In contrast to this result, PDT with 100 J/cm2 did not result in increased levels of active TGF-beta (8.1 +/- 3.5 pg), despite having similar levels of total TGF-beta. Consequently, the conditioned media of SMCs that had 100 J/cm2 PDT did not inhibit endothelial cell mitogenesis as compared with the conditioned media of SMCs with mechanical injury and 10 J/cm2 PDT (p < 0.0002). CONCLUSIONS: This report describes two novel findings: (1) injury to SMCs in vitro induces the conversion of biologically latent TGF-beta to active TGF-beta; and (2) the therapeutic PDT dose interferes with this injury activation process. This study substantiates the concept of local cytokine inhibition by PDT in a biologic system and provides new insights into the mechanisms of PDT-mediated inhibition of experimental IH. PMID- 9201166 TI - Impact of activated protein C resistance on general vascular surgical patients. AB - PURPOSE: The prevalence of activated protein C resistance (APCR) and associated thrombotic morbidity among patients who undergo arterial reconstruction were investigated. METHODS: Preoperative assays for functional APCR and factor V (Leiden) mutation were performed on 262 patients who underwent arterial reconstructions that consisted of cerebrovascular surgery (109), aortic or iliofemoral procedures (76), or infrainguinal bypass procedures (77). Patients were monitored for thrombotic complications during the postoperative period. RESULTS: Depending on the stringency of the definition used, functional APCR was detected in 10.6% to 22.0% of patients tested. Factor V (Leiden) was found in 5.3% of patients. Thrombotic morbidity consisting of myocardial infarction, cerebrovascular event, or graft thrombosis occurred in 9.9% of patients, who were followed-up for a mean of 4.8 months. No significant overall correlations were found between APCR and thrombotic morbidity. Subgroup analysis revealed significant associations between functional APCR and total early postoperative thrombotic complications and early graft failure, and between factor V (Leiden) and early cerebrovascular events and late graft thrombosis (p < 0.03). CONCLUSIONS: Functional APCR is somewhat more prevalent among general vascular surgical patients than in the general population, but factor V (Leiden) is no more prevalent. APCR is not a prominent cause of thrombotic morbidity in contemporary vascular surgery. Nonetheless, it is a sufficiently important potential contributor to morbidity among some subgroups to warrant selective testing and directed therapy pending further study. PMID- 9201167 TI - The role of integrins in saphenous vein vascular smooth muscle cell migration. AB - PURPOSE: Smooth muscle cell (SMC) migration is an essential feature of the intimal hyperplastic process that so frequently limits the patency of vascular reconstructions. The purpose of this investigation was to evaluate the effect of a series of integrins, or cell surface receptors that mediate cellular attachment, on platelet-derived growth factor (PDGF) and extracellular matrix (ECM) protein-induced migration of human SMCs. METHODS: Immunofluorescence staining was used to search for various integrins and subunits on the surface of SMCs derived from human saphenous vein. Chemotaxis and haptotaxis of SMCs to various matrix proteins and PDGF were assayed using a 48-well microchemotaxis chamber in the presence or absence of antibodies that blocked the function of these integrins. RESULTS: Several subunits (beta 1, alpha 2, alpha 5) and one integrin (alpha v beta 3) were identified in saphenous vein SMCs. The beta 1 integrin antibody inhibited chemotaxis to collagen I and IV, laminin, and PDGF. The alpha 2 integrin antibody inhibited collagen I and IV, and laminin-induced chemotaxis. The alpha 5 integrin antibody had no effect on SMC migration. The alpha v beta 3 integrin antibody inhibited chemotaxis to PDGF but not to the ECM proteins. CONCLUSIONS: Integrins are necessary for SMC migration induced by PDGF and ECM proteins. The integrin or subunits responsible for facilitating migration varies with the stimulant. Agonists designed to inhibit integrin function might be used to suppress SMC migration and suppress the formation of intimal hyperplasia. PMID- 9201168 TI - Diabetes mellitus: a risk factor for carotid endarterectomy? AB - PURPOSE: Symptomatic cerebrovascular disease is more common in patients who have diabetes mellitus than in the nondiabetic population, even when matched for associated risk factors. Although the safety and efficacy of carotid endarterectomy has been established by NASCET and ACAS, several small studies have noted an increased rate of perioperative neurologic morbidity in patients with diabetes. METHODS: Data for all patients who underwent carotid endarterectomy at a single institution from Jan. 1990 to Dec. 1995 were prospectively entered into a computerized vascular registry and form the basis of this report. RESULTS: Of 732 carotid endarterectomy procedures performed, 284 (39%) were performed in patients who had diabetes mellitus. Patients with diabetes and without diabetes were matched for clinical presentation (diabetic patients, 45% asymptomatic; nondiabetic patients, 43%) and internal carotid artery percent stenosis (86.6% +/- 10.6% vs 86.4% +/- 10.6%). Patients with diabetes were younger at presentation than patients without (68.8 +/- 8.5 years vs 70.9 +/- 8.5 years; p < 0.005) and were more likely to have a history of coronary artery disease (53% vs 45%; p = 0.04). The mean total length of stay was 6.1 days for patients with diabetes and 4.8 days among patients without (p = 0.01). An adverse postoperative cardiac event (myocardial infarction, congestive heart failure, or arrhythmia) occurred in nine patients with diabetes (3.2%) and in five nondiabetic patients (1.1%; p < 0.05). By logistic regression analysis, however, diabetes was not an independent risk factor for a postoperative cardiac event (p = 0.28). There were 11 perioperative neurologic events (eight cerebrovascular accidents, three transient ischemic attacks) during the entire period (1.5%), of which six were among diabetic patients (2.1%) and five among nondiabetic patients (1.1%; p = NS). Of the eight cerebrovascular accidents, three occurred in diabetic patients (1.0%) and five in nondiabetic patients (1.1%; p = NS). The total operative mortality rate was 0.3% (diabetic patients, 1 of 284, 0.35%; nondiabetic, 1 of 447, 0.2%). CONCLUSIONS: Patients with diabetes who undergo carotid endarterectomy are more likely to have coexisting cardiac disease, which may contribute to a higher incidence of postoperative cardiac morbidity. Diabetes mellitus alone, however, is not a risk factor for postoperative cardiac morbidity in patients who undergo carotid surgery. In addition, carotid endarterectomy may be safely performed in patients with diabetes with neurologic morbidity and mortality rates that are comparable with those of the nondiabetic population PMID- 9201169 TI - Vascular surgery and the Resource-based Relative Value Scale five-year review. AB - PURPOSE: The first 5-year review of the Medicare Resource-based Relative Value Scale (RBRVS) work values (RVUs) began in 1995, and adjustments became effective January 1, 1997. This report summarizes the methods used by The Society for Vascular Surgery (SVS) and the International Society for Cardiovascular Surgery, North American Chapter, (ISCVS-NA) Joint Council Government Relations Committee (GRC) to evaluate vascular surgery work RVUs and the results that were achieved. METHODS: The GRC performed a work study to determine accurate skin-to-skin operative times for typical vascular and nonvascular operations. These were compared with the original Harvard/Hsiao time estimates and intraservice work per unit time (IWPUT) values that had been used to determine work RVUs. For most vascular procedures the current operative times were longer than the original Harvard estimates, resulting in calculated IWPUTs substantially less than the Harvard values. This lack of correspondence was not identified in the nonvascular procedures, where operating room times and IWPUT values were more consistent with Harvard data. These study results were then used to support compelling evidence arguments in a petition to the Health Care Financing Administration (HCFA) that identified vascular surgery as being undervalued in the RBRVS. Nine commonly performed vascular procedures were cited for review in the 5-year update, and five distinct work analysis methods were used to justify each recommended RVU increase. These techniques included a standardized survey from the American Medical Association (AMA)/Specialty Society Relative Value Update Committee (RUC), a work calculation using accurate intraservice times and appropriate IWPUT values, and an evaluation and management (E&M) building-block approach. RESULTS: The RUC met throughout 1995 to assess codes submitted for review, and recommendations were forwarded to HCFA. The Notice of Proposed Rule Making (NPRM), which contained HCFA's preliminary RVU determinations, was released in May 1996. RVU increases from 11.5% to 44.6% were proposed for the nine vascular services cited by the SVS/ISCVS-NA. Also included were two increases and two reductions in less-common vascular operations. Of far greater overall fiscal import, HCFA proposed substantial increases in the work RVU for all E&M except that performed within global surgical packages. The SVS/ISCVS and most other surgical societies appealed HCFA's proposal regarding E&M. The Final Rule for the 1997 Medicare Fee Schedule was published late in 1996. CONCLUSIONS: The Final Rule upheld the 11 vascular work value improvements and the E&M increases that excluded global service packages. Because most surgical E&M is performed within 10- or 90-day global periods, the E&M ruling will produce an estimated annual $2.5 billion shift from surgical to nonsurgical specialties. Because the overall fiscal impact of the 5-year review was mandated to be budget-neutral, HCFA imposed an 8.3% reduction in the work payment of every service in Part B of the Medicare program, primarily to compensate for the increased nonsurgical E&M payments. The net fiscal impact of the 5-year review for vascular surgery has been estimated at +0.5%. PMID- 9201170 TI - Regarding "Statement regarding carotid angioplasty and stenting". PMID- 9201171 TI - Regarding "Infected thrombophlebitis of the right internal jugular vein". PMID- 9201173 TI - Cord blood banking and transplantation. PMID- 9201172 TI - Prospective randomized trials of carotid endarterectomy with primary closure and patch reconstruction: the problem is power. PMID- 9201174 TI - Multiple sclerosis, multiple genes. PMID- 9201175 TI - Rationing and the objectives of health care. PMID- 9201176 TI - Factors in accessibility of general practice in rural Australia. AB - OBJECTIVE: To ascertain the importance rural Australians attribute to different factors of accessibility in their decision to consult a general practitioner. DESIGN: Survey by interview or delivery-and-collection questionnaire (participant's choice) based on the method of paired comparisons. SETTING AND PARTICIPANTS: All residents of 10 small rural communities in north-west New South Wales aged over 16 years in July and August 1996. MAIN OUTCOME MEASURES: Rank order and relative importance of residents' preferences for choosing to consult a particular doctor. RESULTS: Social accessibility or acceptability considerations were more important than geographical proximity in the choices of rural residents to consult a particular doctor. Elderly people, in particular, attributed most significance to acceptability and continuity of care. Geographical proximity ranked most highly for young and middle-aged people and men living in isolated communities. CONCLUSIONS: For rural inhabitants, geographical distance is not the sole or even the most important determinant in their choice of general practice care; rather, they will seek the services of a GP with whom they feel comfortable. Incentives programs designed to recruit and retain more GPs in rural practice must acknowledge the importance of attracting acceptable doctors. This requires that rural doctors acquire suitable clinical and communication skills to meet the diverse needs of their patients, as well as an understanding of rural culture. PMID- 9201177 TI - Characteristics of incompletely excised basal cell carcinomas of the skin. AB - OBJECTIVES: To determine the proportion of basal cell carcinomas (BCCs) treated by excision biopsy that extended to the margins of surgical excision (incompletely excised tumours) and to identify their characteristics. DESIGN: Case series of BCCs submitted to a single pathologist in the first six months of 1995. SETTING: Rural and metropolitan (Perth) Western Australia. PATIENTS: 268 patients with 353 histologically confirmed BCCs. OUTCOME MEASURES: Age and sex of patients; discipline of referring doctor; anatomical site of BCC; macroscopic features; histological growth pattern; and completeness of excision. RESULTS: Sixteen per cent of BCCs (58/353) extended to the margin of surgical excision. Most of these were situated on the head or neck (43/58; 74%) and were flat (47/58; 81%); a high proportion of incompletely excised BCCs (21/58; 36%) had an infiltrative growth pattern. Recurrent BCCs (28/353; 8%), categorised from the history or because of histologically identified surgical scarring, were even more likely to be flat (26/28; 93%) and to show a microscopic infiltrative growth pattern (18/28; 64%). Seven of the 28 (25%) recurrent BCCs were incompletely excised; all seven were on the head and five had an infiltrative growth pattern. CONCLUSION: Incompletely excised BCCs are those most likely to recur. Because most recurrent tumours are situated on the head and neck and have an infiltrative growth pattern, we recommend that: Pathologists report on the microscopic growth pattern of BCCs as well as on completeness of excision. Clinicians attempt to excise head and neck BCCs with wide margins initially, where possible. Tumours extending to the margin of excision which are infiltrative in pattern and located on the head and neck may be particularly likely to recur, and immediate re excision should be considered in these patients. PMID- 9201178 TI - The risk of transmitting HCV, HBV or HIV by blood transfusion in Victoria. AB - OBJECTIVE: To report the incidence rate of hepatitis B virus (HBV), hepatitis C virus (HCV) and HIV in Victorian repeat blood donors and to derive the residual risk of transmission of the viruses by screened blood transfusion. DESIGN: The interval from the previous whole blood donation was extracted retrospectively from Victorian Red Cross Blood Bank records for each of the 358332 repeat donations given between March 1994 and December 1995. Records of repeat donors found positive for the viruses in this period were traced to the previous seronegative donation and accepted if screened by the same test. For each virus, the number of previous donations screened by the same test was calculated and the sum of all donation intervals used to derive the incidence of infection in the repeat donor population. Published intervals after infection (when a donation can be infective although seronegative) were used to calculate the risk of release of a seronegative unit which would be infective. PARTICIPANTS AND SETTING: Homologous blood donors at the Red Cross Blood Bank of Victoria. MAIN OUTCOME MEASURES: Incidence rate of HBV, HCV and HIV in regular blood donors and risk of infective donations being seronegative. RESULTS: The incidence of infection in repeat donors was: HBV: 1.67 per 100000 person-years; HCV: 1.89 per 100000 person years; and HIV: 1.31 per 100000 person-years. The risk of a seronegative repeat donation being infective was: HBV: 2.71 per million donations (adjusted to 6.45 to account for viraemias which remain seronegative); HCV: 4.27 per million donations; and HIV: 0.79 per million donations. CONCLUSION: The risk of transmitting HCV, HBV or HIV by repeat blood donors is low and compares favourably with overseas data. Repeat donors have an incidence rate of HIV and HBV comparable to that of the general population, but the incidence rate of HCV is lower for repeat donors than in the general population. PMID- 9201179 TI - A red-back spider bite in a lymphoedematous arm. AB - We administered a record eight ampoules of antivenom over one week to a 55-year old woman with a red-back spider bite on her chronically lymphoedematous right arm. She had severe local signs of envenomation but relatively little systemic effect. The final antivenom dose, injected subcutaneously at the bite site, had a good effect. PMID- 9201180 TI - What are the risks of diagnostic medical radiation? AB - It is both ethically and economically desirable to restrict the use of diagnostic medical radiation to only those who will benefit from it. However, patients should not refuse diagnostic tests based on an exaggerated estimation of the risks because most of these tests involve low doses of radiation. It is probable that the risks derived from studies of the atomic bomb survivors, who were exposed to high doses of radiation, overestimate the risks at low doses. No evidence of thyroid cancer, leukaemia or non-Hodgkin's lymphoma has been found in patients exposed to diagnostic levels of ionising radiation. For most diagnostic tests, the risks arising from the radiation exposure are too small to be observed and the benefits will almost always outweigh the risk. PMID- 9201181 TI - Clinical practice guidelines: to what end? AB - The move to develop clinical practice guidelines in Australia is gaining momentum as part of a national approach to improving the quality of clinical practice. The National Health and Medical Research Council has published a "guidelines for guidelines". While it has also produced guidelines for nine specific clinical topics, it is now passing this role to professional organisations, such as clinical colleges and learned societies, and reverting to an overseeing, facilitating and credentialling role. PMID- 9201182 TI - New guidelines for management and prevention of meningococcal disease in Australia. Meningococcal Disease Working Party of the National Health and Medical Research Council. AB - The incidence of invasive meningococcal disease in Australia has increased over the past decade, and in April 1997 the National Health and Medical Research Council published guidelines for management of patients with meningococcal disease and their contacts. These guidelines emphasise the need for immediate intravenous antibiotic treatment of patients with suspected meningococcal disease, before transfer to hospital or lumbar puncture. When possible, blood for culture should be collected before antibiotic therapy, if this does not delay treatment. PMID- 9201183 TI - Anaesthesia--150 years strong. PMID- 9201184 TI - The chest wall and pleural space. PMID- 9201185 TI - The effectiveness of donor selection for reducing the risk of HCV, HBV and HIV in new blood donors. PMID- 9201187 TI - Top school marks don't necessarily make top medical students. PMID- 9201186 TI - "Christmas eye". PMID- 9201188 TI - Selecting Australian doctors of the future. PMID- 9201189 TI - The rural medical crisis. PMID- 9201190 TI - Blood lead concentrations and iron status of preschool children from low income families. PMID- 9201191 TI - Circumcision blues. PMID- 9201192 TI - Anogenital warts. PMID- 9201194 TI - Telemedicine: here to stay. PMID- 9201193 TI - Silicone breast implants: implications for society and surgeons. PMID- 9201195 TI - An inexpensive "orthosis" for plantar fasciitis. PMID- 9201197 TI - Recent advances in managing non-small-cell lung cancer: 2. Surgery. AB - Surgery can be curative in non-small-cell lung cancer, but has only a minor role in small-cell lung cancer. About two-thirds of patients with non-small-cell lung cancer present with advanced disease that cannot be treated surgically. Lymph node involvement alone does not indicate inoperable or incurable cancer. Lobectomy in patients with Stage I disease results in a five-year survival rate of 70%. Lobectomy or pneumonectomy with resection of involved nodes in patients with Stage II disease has a five-year survival rate of 40%-45%. Carefully selected patients with Stage IIIA disease may benefit from surgical treatment. PMID- 9201196 TI - Recent advances in managing non-small-cell lung cancer: 1. Clinical aspects of diagnosis and staging. AB - Lung cancer is the leading cause of death from cancer in Australian adults. Over 90% of lung cancers are due to exposure to tobacco smoke. Female smokers are more susceptible to developing lung cancer than male smokers. The overall survival of non-small-cell lung cancer is 13%, having increased from 6% over the past 30 years. The development of new, unexplained respiratory or systemic symptoms in a heavy smoker or ex-smoker should raise the suspicion of lung cancer. Early detection and consideration of tumour resection for all operable tumours are the main strategies for improving the cure rate of non-small-cell lung cancer at present. Reducing the prevalence of smoking remains the best method for reducing lung cancer deaths. PMID- 9201198 TI - Recent advances in managing non-small-cell lung cancer: 3. Radiation therapy. AB - The most important role of radiation therapy in NSCLC is the palliation of symptoms due to intrathoracic and metastatic disease. Short courses of treatment appear to be as effective as protracted courses. In patients with good performance status and no evidence of distant metastases, high dose radiation therapy (radical radiation therapy) provides short term survival benefits. Up to a quarter of stage I patients may survive for five years. Combinations of chemotherapy and radiation therapy appear to be more effective than radiation therapy alone, and sufficient evidence now exists to support combined radiation therapy and platinum-based chemotherapy as the best standard of care for patients with good performance status and unresectable disease. PMID- 9201199 TI - Recent advances in managing non-small-cell lung cancer: 4. Chemotherapy of metastatic cancer. AB - Chemotherapy has been shown to prolong survival and to be cost effective in the palliative treatment of non-small-cell lung cancer. A meta-analysis of 11 randomised trials has found a small benefit from combination chemotherapy in advanced NSCLC, increasing one-year survival rates from 5% to 15%. New combination regimens, especially carboplatin-paclitaxel, show higher response rates in selected patients, and promise improved survival rates, but this has yet to be confirmed in randomised trials. PMID- 9201200 TI - Recent advances in managing non-small-cell lung cancer: 5. Endobronchial palliative therapy. AB - A variety of treatments can be effective palliative therapy of symptoms of airway obstruction caused by endobronchial tumour: Nd-YAG laser resection may be indicated for tumours that are relatively short in length, situated in trachea, mainstem or proximal lobe bronchi. Cryotherapy may be an alternative to laser resection. Endobronchial brachytherapy (precise delivery of radiation to an endobronchial tumour via a catheter loaded with iridium 192) may be indicated in patients with endobronchial mural disease or tumours that extend beyond the bronchial wall. Endobronchial stents. Randomised trials are required to determine the relative merits of these treatments and the optimal management of endobronchial complications of lung cancer. PMID- 9201201 TI - Creutzfeldt-Jakob disease. PMID- 9201202 TI - Screening for prostate cancer: what do general practitioners think? AB - AIM: To determine how general practitioners in New Zealand view screening for prostate cancer and the extent to which this is undertaken in general practice. METHOD: A questionnaire survey of a random sample of 500 general practitioners. RESULTS: Completed questionnaires were received from 317 of an eligible sample of 462. Approximately 50% believed digital rectal examination (DRE) and prostate specific antigen (PSA) were effective screening tests for prostate cancer and that asymptomatic men should be screened; 40% believed all men aged 50 years or more should be screened using either DRE or PSA. The majority of the general practitioners currently screen at least some of the men aged 50 years or more on their lists using DRE or PSA regardless of beliefs about the efficacy of the tests. The results also indicated that significantly more general practitioners in the age groups 50-59 years and 60 years and over believed asymptomatic patients should be screened with DRE or PSA. CONCLUSION: Despite the absence of evidence to support screening for prostate cancer using DRE or PSA and the increasing number of professional organisations releasing guidelines and statements to that effect, the majority of the general practitioners who participated in this survey are screening some of their patients aged 50 years or more using DRE and/or PSA. PMID- 9201203 TI - A comparison of two pertussis epidemics in Auckland. AB - AIM: To determine if the addition of the 6 week dose of pertussis vaccine in 1984 was associated with any change in the hospitalisation rate for children with pertussis and the higher hospitalisation rates for Maori and Pacific Islander children with pertussis. METHODS: DESIGN: Population based study of pertussis hospitalisations using a retrospective chart review of hospitalisation data for children during the 1991 epidemic, which was compared to previously published data from the 1982 epidemic. SETTING: Princess Mary and Middlemore hospitals, Auckland. SUBJECTS: Children aged 0-14 years resident in metropolitan Auckland and hospitalised in Auckland during 1982 or 1991 with pertussis. MEASUREMENTS: Hospitalisation rates were calculated as number of children with a discharge diagnosis of pertussis per 1000 children aged 0-14 years based on 1981 and 1991 census data. 1982 data were converted to person-years as published report was for an 8 month period. Hospitalisation rates were compared as a relative risk (RR) of hospitalisation in 1991 versus 1982. RESULTS: There were 84 cases during 8 months in 1982 and 66 cases in 1991. Rates of hospitalisation by ethnic group; in 1982 were 0.24 Other/European (OE), 1.98 Maori (M), 1.37 Pacific Islander (PI); and in 1991 were 0.22 OE, 0.51 M, 0.40 PI. Compared to 1982 the relative risk of hospitalisation in 1991 adjusted for ethnicity was 0.43 (CI 0.33, 0.58, p < 0.0001). Compared to 1982 there was a significant reduction in the hospitalisation rate in 1991 for M (RR = 0.26, CI 0.16, 0.43, p < 0.0001); and PI children (RR = 0.29, CI 0.16, 0.54, p < 0.0001); but not for OE children (RR = 0.91, 95% CI 0.57, 1.46, p = 0.70). CONCLUSIONS: There was a significant reduction in the rate of hospitalisation for pertussis in 1991 compared to 1982. This reduction in hospitalisation rate was due to a reduction in rates for Maori and Pacific Islander children. PMID- 9201204 TI - Baby feeding: the thoughts behind the statistics. AB - AIM: To investigate attitudes towards baby feeding and to identify reasons why women stop breast feeding. METHOD: A series of six focus groups were held with thirty eight mothers with babies aged between 3 and 18 months, who had been breast and/or bottle fed. RESULTS: The discussions identified a number of significant themes. Decisions on baby feeding were made before birth. Women felt under considerable pressure to breast feed and felt guilty about bottle feeding. Information available about baby feeding was generally inconsistent, unrealistic and incomplete although all women were well informed about the benefits of breast feeding. Most women found breast feeding more difficult than anticipated and needed more help with common problems. A number of difficulties were identified with bottle feeding. Those women who enjoyed breast feeding were most likely to continue. The best support for breast feeding came from other mothers and supportive partners. Ceasing breast feeding was difficult for some women. CONCLUSION: Exploring mothers' attitudes to breast feeding highlighted the need for non judgemental attitudes to baby feeding and consistent information and support on both breast and bottle feeding. Duration of breast feeding is likely to be improved if problems can be addressed. A larger, more detailed prospective study would more accurately identify problem areas and suggest ways of solving them. PMID- 9201206 TI - Octreotide for the treatment of sulphonylurea induced hypoglycaemia in type 2 diabetes. PMID- 9201205 TI - Susceptibility patterns of bacterial isolates from intensive care and haematology/oncology patients in New Zealand. AB - AIMS: To determine the current susceptibility pattern of bacterial isolates from intensive care and haematology/oncology patients in New Zealand. METHOD: Over a 6 month period 417 consecutive clinically relevant bacterial isolates from intensive care and haematology/oncology patients from seven New Zealand hospitals had their susceptibility to multiple antimicrobial agents determined by the agar plate dilution method. Methicillin resistant staphylococci were not included. RESULTS: Of the 417 isolates, 224 (54%) were gram negative and 193 were gram positive. Predominant species/groups were: Escherichia coli 63 (15%), Enterobacter spp 26 (6%), other Enterobacteriacae 41 (10%), Pseudomonas aeruginosa 42 (10%), Staphylococcus aureus 111 (27%), coagulase negative staphylococci 30 (7%), Streptococcus spp 31 (7%), and Enterococcus spp 19 (5%). Isolate sources were: respiratory tract, 170 (41%); cutaneous sites, 81 (19%); blood, 64 (15%); and urine 63 (15%). Resistance was uncommon amongst staphylococci, streptococci, enterococci, and H influenzae. No vancomycin resistant or beta-lactamase-positive enterococci were encountered. For different groups of enteric gram negative bacilli: amoxycillin and amoxycillin-clavulanic acid resistance was common, 46-93% and 24-85% respectively; cefpirome was the most active cephalosporin; aminoglycoside resistance was uncommon; and no isolate possessed extended spectrum beta-lactamase. For P aeruginosa: most isolates were susceptible to cefpirome and ceftazidime, and aminoglycoside resistance was uncommon. CONCLUSION: Gram positive bacteria make up a higher proportion of isolates than in a similar European study. At present New Zealand does not have widespread resistance amongst common isolates. Several agents currently available in New Zealand provide adequate cover for commonly encountered pathogens. The choice of which agent to choose therefore rests more with their purchase and administration costs, as well as safety and efficacy data than simply susceptibility data alone. PMID- 9201207 TI - Visit to Qui Nhon, Vietnam, by New Zealand medical team. PMID- 9201208 TI - Te Waikato: sanatorium for consumptives. PMID- 9201210 TI - The practice of medicine. PMID- 9201211 TI - Acute psychiatric bed availability. PMID- 9201209 TI - Measles control. PMID- 9201212 TI - Entry of mouse hepatitis virus into cells by endosomal and nonendosomal pathways. AB - OBLV60 is an acid-dependent syncytium-forming variant isolated from OBL21 cells persistently infected with the pH-independent mouse hepatitis virus (MHV)-4 strain. The fusion activity of OBLV60 can be strictly regulated by controlling pH and thus provides the means to definitively examine the entry of MHV into cells by endosomal and nonendosomal pathways. Shortly after high multiplicity infection, both MHV-4 and OBLV60 were detected by electron microscopy in endosomal vesicles and were recovered from lysates of cells treated with proteinase K to remove extracellular virus. For OBLV60, but not MHV-4, exposure to lysosomotropic compounds early in infection prevented viral penetration and significantly reduced viral yields. These results suggested that both MHV-4 and OBLV60 utilized the endosomal route of entry into cells, but that MHV-4 did not require acidification of endosomal vesicles. Studies on the entry of virus through fusion at the cell surface were performed by briefly exposing surface bound OBLV60 to a fusion-permissive pH under conditions that prevent endocytic entry. Acid treatment of surface-bound OBLV60 caused a significant increase in the yields of virus produced in cultures of fusion-sensitive Sac- or DBT cells, demonstrating entry of virus by fusion at the cell surface. No measurable increase in virus production was detected with acid treatment of OBLV60 bound to OBL21 cells, suggesting that entry at the cell surface does not occur in these cells, which are resistant to MHV-induced syncytia formation. These results raise interesting questions concerning how mechanisms of MHV entry influence the selection of fusion variants. PMID- 9201213 TI - Genetic analysis of internal ribosomal entry site on hepatitis C virus RNA: implication for involvement of the highly ordered structure and cell type specific transacting factors. AB - Hepatitis C virus (HCV) carries an internal ribosomal entry site (IRES) within the 5' portion of the RNA. To identify structures that influence efficiency of the translation initiation, relative activities of modified IRESs were examined by using engineered bicistronic mRNAs, between the two cistrons of which various mutant IRESs were inserted. An IRES derived from genotype 2b is at least two times more efficient than one from genotype 1b in cultured cells. Activity ratios of genotype 2b IRES to 1b IRES differ in magnification among cultured cells, suggesting the difference in assortment of IRES-related host factors among individual cell types. Recombinant IRESs between the genotypes show similar or higher activities compared with 2b IRES in cell-free systems and show intermediate activities in cultured cells. Patterns of relative activities of those IRESs indicate that the IRES activity is not regulated by defined structure(s), although a cluster of different nucleotides is observed in the genome region of nucleotides 176-224 between the two alleles. The results suggest that a highly ordered structure formed by the entire 5' portion of the RNA is important for the IRES activity. The 5' border of HCV IRES was examined by using a series of deletion RNAs in various systems. The results strongly suggest that the border resides between nucleotide positions 28 and 45. Patterns of relative activities of the deletion IRESs differ in translation systems or cell types. These results imply that interactions of HCV RNA with the related transacting factor(s) may differ in the translation systems or cell types. PMID- 9201214 TI - The genome of molluscum contagiosum virus: analysis and comparison with other poxviruses. AB - Analysis of the molluscum contagiosum virus (MCV) genome revealed that it encodes approximately 182 proteins, 105 of which have direct counterparts in orthopoxviruses (OPV). The corresponding OPV proteins comprise those known to be essential for replication as well as many that are still uncharacterized, including 2 of less than 60 amino acids that had not been previously noted. The OPV proteins most highly conserved in MCV are involved in transcription; the least conserved include membrane glycoproteins. Twenty of the MCV proteins with OPV counterparts also have cellular homologs and additional MCV proteins have conserved functional motifs. Of the 77 predicted MCV proteins without OPV counterparts, 10 have similarity to other MCV proteins and/or distant similarity to proteins of other poxviruses and 16 have cellular homologs including some predicted to antagonize host defenses. Clustering poxvirus proteins by sequence similarity revealed 3 unique MCV gene families and 8 families that are conserved in MCV and OPV. Two unique families contain putative membrane receptors; the third includes 2 proteins, each containing 2 DED apoptosis signal transduction domains. Additional families with conserved patterns of cysteines and putative redox active centers were identified. Promoters, transcription termination signals, and DNA concatemer resolution sequences are highly conserved in MCV and OPV. Phylogenetic analysis suggested that MCV, OPV, and leporipoxviruses radiated from a common poxvirus ancestor after the divergence of avipoxviruses. Despite the acquisition of unique genes for host interactions and changes in GC content, the physical order and regulation of essential ancestral poxvirus genes have been largely conserved in MCV and OPV. PMID- 9201215 TI - Mutation rate of GB virus C/hepatitis G virus over the entire genome and in subgenomic regions. AB - A patient on maintenance hemodialysis was infected with a recently discovered putative non-A to -E hepatitis virus designated GB virus C (GBV-C) or hepatitis G virus (HGV) by transfusion. The viral isolate was recovered from the patient soon after she turned positive for GBV-C/HGV RNA in serum (GS185) and 8.4 years thereafter (GS193), and the entire nucleotide sequences were determined. They both had a genomic length of 9391 nucleotides with a defective C gene made of only 42 nucleotides. Between GS185 and GS193, 31 (0.33%) nucleotides were different, which changed 5 (0.18%) of the encoded 2842 amino acids. Thus, GBV C/HGV was estimated to have a mutation rate of 3.9 x 10(-4) base substitutions per site per year. Nucleotide conversions were distributed over subgenomic regions, except in the 5' untranslated region of 552 nucleotides and a defective short C gene, which were conserved in sequence. The change in the putative envelope genes (E1 and E2) was no different from that in the entire genome with only 6 (0.35%) nucleotide substitutions among the 1730, just 1 of which induced an amino acid conversion. Taken along with the comparison of the two isolates with the reported five GBV-C or HGV isolates, these results indicate that GBV C/HGV would not have hypervariable regions and would use a strategy for viral persistence that is different from immune escape. PMID- 9201216 TI - Mutations in CR1 of E1A 12S yield dominant negative suppressors of immortalization and the lytic cycle. AB - The Adenovirus 5 E1A 12S gene is responsible for the establishment of immortalization of primary cells by Adenovirus. We have identified two mutants of 12S (30K and NTdl814), which, when coexpressed with wild-type 12S in primary baby rat kidney cells, were capable of suppressing the immortalizing function of the wild-type 12S gene, even when the mutant proteins were expressed at levels lower than wild type. 30K and NTdl814 did not affect the ability of the coexpressed 12S to activate the cell cycle, but have a suppressive effect on 12S-induced DNA synthesis and proliferation at late times in the immortalization pathway. Both the dominant negative mutants have a deletion in conserved region (CR)1 in the first exon of E1A, which encompasses one of the pRb-family binding regions. However, the mutants did not effect the binding of cellular proteins to full length 12S. A suppressive effect on wild-type 12S was not observed with mutants that have lost any other region or function. In addition, expression of 30K, which is equivalent to the protein encoded by the 10S mRNA of E1A, inhibited E1A function in lytic cycle. Thus, loss of the CR1 seems to be a prerequisite for a mutant to have a dominant negative effect on E1A functions. PMID- 9201217 TI - RNA synthesis by the brome mosaic virus RNA-dependent RNA polymerase: transition from initiation to elongation. AB - The initiation and elongation phases of (-)-strand RNA synthesis in vitro by the brome mosaic virus RNA-dependent RNA polymerase (RdRp) are differentially sensitive to inhibitors. In an attempt to characterize further the transition RdRp makes from initiation to elongation, we determined the conditions needed to pause the ternary complex and complete only one round of RNA synthesis. During the transition we were able to discern step-wise increases in the affinity of RdRp for RNA by measuring sensitivity to heparin and competition for RdRp by an alternative template. Three distinct stability levels of RdRp-template interactions were found. The first stable RdRp-RNA complex was observed when RdRp bound to the RNA template. A further increase occurred when RdRp synthesized the first phosphodiester bond. A final increase occurred upon formation of between 3 and 13 phosphodiester bonds. After this last transition, RdRp appeared to be tightly committed to the template RNA. These results are analogous to the mechanism of action of DNA-dependent RNA polymerases and are relevant to protein RNA interaction and template switching by an RdRp. PMID- 9201218 TI - Experimental measles. I. Pathogenesis in the normal and the immunized host. AB - An animal model to study measles pathogenesis and the correlates of protective immunity was established using rhesus monkeys. A measles isolate, obtained during an epidemic of measles in the primate colony at the University of California, Davis, was passaged through rhesus monkeys and amplified in rhesus mononuclear cells to create a pathogenic virus stock. Sequence analysis of the nucleoprotein and hemagglutinin genes of this isolate revealed strong homology with the Chicago 89 strain of measles virus. Conjunctival/intranasal inoculation of juvenile rhesus monkeys with this virus resulted in skin rash, pneumonia, and systemic infection with dissemination to other mucosal sites and to the lymphoid tissues. Inflammation and necrosis occurred in the lungs and lymphoid tissues and many cell types were infected with measles virus on Day 7 postinoculation (p.i.). The most commonly infected cell type was the B lymphocyte in lymphoid follicles. Measles antigen was found in follicular dendritic cells on Day 14 p.i. In contrast to naive monkeys infected with measles virus, animals vaccinated with the attenuated Moraten strain did not develop clinical or pathologic signs of measles after challenge. However, moderate to marked hyperplasia occurred in the lymph nodes and spleen of a vaccinated animal on Day 7 after pathogenic virus challenge, suggesting that an effective measles vaccine limits but does not prevent infection with wild-type measles virus. PMID- 9201220 TI - A conserved internal hydrophobic domain mediates the stable membrane integration of the dengue virus capsid protein. AB - The mature flavivirus capsid protein (virion C) is commonly thought to be free in the cytoplasm of infected cells and to form a nucleocapsid-like complex with genomic RNA in mature virus particles. There is little sequence conservation among flavivirus virion C proteins, but they are similar in size (e.g., 99 amino acids [aa] for the dengue-4 [DEN4] C) and in bearing a net positive charge. In addition, we noted that C contained a conserved internal hydrophobic segment (spanning aa 45-65 in the DEN4 C). Results of in vivo expression and in vitro translation of wt and mutant forms of the DEN4 virion C demonstrated that the conserved internal hydrophobic segment in the DEN C functioned as a membrane anchor domain. Signal peptide function of this segment was also suggested by its requirement for the entry of C into membranes. Virion C was integrated in membranes in a "hairpin" conformation; positively charged segments amino- and carboxy-terminal to the hydrophobic signal-anchor segment were accessible to protease digestion in the "cytoplasm." The net positive charge in the amino terminal extramembraneous portion of C (aa 1-44) was one determinant of the hairpin membrane orientation; a conserved positively charged residue within the hydrophobic segment (Arg-54 in the DEN4 C) was not. PMID- 9201219 TI - In vivo longitudinal analysis of a dominant TCR repertoire selected in human response to influenza virus. AB - Recent studies have demonstrated biased usage of TCR V beta 17 and a high degree of diversity in J beta usage within the influenza virus matrix epitope (M.58-66) specific CTL response. In contrast, in the course of a study on the cellular response to influenza A virus, we found preferential usage of V beta 17-J beta 2.2 rearrangement in an individual with an unexpectedly high number of CTL precursors (CTLp). We took advantage of such situation to study the longitudinal repertoire of the CD8+ T cell precursors. By limiting dilution analysis combined with the use of a clonotypic primer corresponding to the CDR3 region of this matrix-specific TCR V beta chain, the influenza-specific CTLp were shown to be stable for a period of 6 years. Overall, our results show that virus-specific CTLp can be directly monitored in vivo by molecular fingerprinting without in vitro restimulation. These findings might be extremely important for evaluation of the specific immune response to a given human pathogen. PMID- 9201221 TI - Attenuation of B5R mutants of rabbitpox virus in vivo is related to impaired growth and not an enhanced host inflammatory response. AB - The rabbitpox virus (RPV) B5R protein, synthesized late in infection, is found as a 45-kDa membrane-associated protein of the envelope of infectious extracellular enveloped virus (EEV) and as a 38-kDa protein secreted from the cell by a process independent of morphogenesis. The protein is not found associated with intracellular mature virus (IMV). Deletion of the gene attenuates the virus (RPV delta B5R) in animals (mice and rabbits), has relatively little effect on formation of IMV, prevents EEV formation in some but not all cells, and leads to a reduced host range. Analysis of the sequence of the protein suggests relatedness to factor H of the complement cascade. Collectively, these observations suggest that attenuation of the virus in vivo could be linked to an inhibition of the inflammatory response, a deficiency in growth, or both. In this report we have analyzed the behavior of RPV delta B5R in infected mice and rabbits and conclude that attenuation of the mutant virus likely results from simple failure to grow within the infected animal and that the inflammatory response probably contributes little to the observed attenuation. PMID- 9201222 TI - SV40 and adenovirus may act as cocarcinogens by downregulating glutathione S transferase expression. AB - We have discovered a novel function of the SV40 T antigen and the adenovirus E1A proteins: the ability to downregulate the endogenous expression of an important detoxification enzyme, glutathione S-transferase alpha (GST alpha). GST alpha mRNA is much less abundant in rat and human cells that express SV40 T antigen than in the parental cell lines. This GST alpha downregulation does not require expression of SV40 small t antigen or complex formation between large T antigen and p53, p300, or the pRb family of proteins. As might be predicted, cells that express SV40 T antigen are more sensitive than normal cells to alkylating drugs, which GST alpha is known to detoxify. Finally, GST alpha expression is also downregulated in cells that express the adenovirus E1A proteins. We propose that by downregulating GST alpha expression and inactivating p53 function, SV40 and adenovirus may contribute to the initiation of, or the progression toward, malignancy. Thus, in their quest to establish persistent infections, these viruses may inadvertently make the cellular environment more permissive for tumorigenesis. PMID- 9201224 TI - Cell attachment and mouse virulence of echovirus 9 correlate with an RGD motif in the capsid protein VP1. AB - The recently analyzed sequences of the nonpathogenic prototype strain Hill and the mouse-virulent strain Barty of the human echovirus 9 differ particularly in an insertion coding for an RGD motif at the C-terminus of the capsid protein VP1 in the genome of strain Barty. To investigate molecular determinants of virulence, we generated a panel of recombinant viruses derived from cDNA clones of strains Hill and Barty. In this communication, we show that the mouse pathogenic character of strain Barty correlates with a 310-aa segment including the RGD motif. By mutating the RGD to an RGE tripeptide, the infectivity of the resulting echovirus 9 clones for GMK cells is lost. Furthermore, we could show that synthetic peptides containing the RGD sequence influence binding of mouse virulent echovirus 9 strains to GMK cells, whereas binding of apathogenic strains is not affected. These results suggest that the RGD motif is a significant factor affecting pathogenicity of echovirus 9 strains. PMID- 9201223 TI - Characterization of the lysogeny DNA module from the temperate Streptococcus thermophilus bacteriophage phi Sfi21. AB - Phage phi Sfi21, the only temperate Streptococcus thermophilus phage from our phage collection, showed extensive DNA homology with virulent phages from lytic group I. Southern blot hybridizations demonstrated that the phi Sfi21-specific DNA was clustered in an approximately 6.6-kb-long region, the putative lysogeny module. Sequence analysis and database research identified an integrase within this module; orf 203 with homology to an anonymous orf 258 from the temperate lactococcal phage BK5-T; orf 127 and orf 122 with weak homology to the N- and C terminal parts, respectively, of the cl-like repressor from lactococcal phages Tuc2009 and BK5-T; orf 75 with homology to a repressor protein from lambdoid phage 434 and an anti-repressor ant with homology to phage P1. The molecular arrangement of the predicted orfs in phage phi Sfi21 was very similar to that of the lactococcal phage BK5-T. The transition from phi Sfi21-specific DNA into DNA shared with virulent phages was abrupt and flanked at one side by notable DNA repeats. Sequence analysis identified a holin protein to the left of the lysogeny module. A site-specific deletion of 2.4 kb, which reproducibly transformed phi Sfi21 into a lytic phage, was localized in the lysogeny module. It was flanked at both sides by conspicuous DNA repeats. One repeat region reflected the DNA around the attP site, while the other reflected the putative genetic switch region between repressor and anti-repressor genes. S. thermophilus host Sfi1 transformed with a plasmid containing int and orf 203 showed resistance to superinfection by heterologous phages, but not by the homologous phi Sfi21. Part of the int gene could be deleted without loss of this activity, while a deletion in orf 203 resulted in loss of the phage resistance. We speculate on the possibility of a bipartite immunity system for the control of lysogeny in phi Sfi21. PMID- 9201225 TI - Long sequences in the 5' noncoding region of plum pox virus are not necessary for viral infectivity but contribute to viral competitiveness and pathogenesis. AB - The 5'-terminal 31 nucleotides of the 146-nucleotides-long 5' noncoding region of plum pox potyvirus (PPV) are highly conserved in all the members of the Potyvirus genus. To map the sequences of the 5' noncoding region that are necessary in vivo for infectivity, we have constructed a nested set of substitution and deletion mutants. While we were not able to infect Nicotiana clevelandii plants with full length PPV transcripts bearing mutations in the 5'-terminal 35 nucleotides of the viral genome, the deletion of long sequences located between nucleotides 39 and 145 did not alter either the rate of infection or viral accumulation. Nevertheless, these mutants were not able to compete with the wild-type strain in coinoculation experiments. Plants infected with a PPV mutant that lacked nucleotides 127 to 145 showed a very mild symptomathology; the wild-type symptom severity was recovered after spontaneous second-site mutations. PMID- 9201226 TI - The influenza A virus M2 channel: a molecular modeling and simulation study. AB - The M2 protein of influenza virus forms ion channels activated by low pH which are proton permeable and play a key role in the life cycle of the virus. M2 is a 97-residue integral membrane protein containing a single transmembrane (TM) helix. M2 is present as disulfide-linked homotetramers. The TM domain of M2 has been modeled as a bundle of four parallel M2 helices. The helix bundle forms a left-handed supercoil surrounding a central pore. Residue H37 has been implicated in the mechanism of low-pH activation of the channel. Models generated with H37 in a fully deprotonated state exhibit a pore occluded by a ring of H37 side chains oriented toward the lumen of the pore. Models with H37 in a fully protonated state no longer exhibit such occlusion of the pore, as the H37 side chains adopt a more interfacial location. Extended molecular dynamics simulations with water molecules within and at the mouths of the pores support this distinction between the H37-deprotonated and H37-protonated models. These simulations suggest that only in the H37-protonated model is there a continuous column of water extending the entire length of the central pore. A mechanism for activation of M2 by low pH is presented in which the H37-deprotonated model corresponds to the "closed" form of the channel, while the H37-protonated model corresponds to the "open" form. A switch from the closed to the open form of the channel occurs if H37 is protonated midway through a simulation. The open channel is suggested to contain a wire of H-bonded water molecules which enables proton permeability. PMID- 9201227 TI - Measles virus recognizes its receptor, CD46, via two distinct binding domains within SCR1-2. AB - Measles virus (MV) enters cells by attachment of the viral hemagglutinin to the major cell surface receptor CD46 (membrane cofactor protein). CD46 is a transmembrane glycoprotein whose ectodomain is largely composed of four conserved modules called short consensus repeats (SCRs). We have previously shown that MV interacts with SCR1 and SCR2 of CD46. (M. Manchester et al. (1995) Proc. Natl. Acad. Sci. USA 92, 2303-2307) Here we report mapping the MV interaction with SCR1 and SCR2 of CD46 using a combination of peptide inhibition and mutagenesis studies. By testing a series of overlapping peptides corresponding to the 126 amino acid SCR1-2 region for inhibition of MV infection, two domains were identified that interacted with MV. One domain was found within SCR1 (amino acids 37-56) and another within SCR2 (amino acids 85-104). These results were confirmed by constructing chimeras with complementary regions from structurally similar, but non-MV-binding, SCRs of decay accelerating factor (DAF; CD55). These results indicate that MV contacts at least two distinct sites within SCR1-2. PMID- 9201228 TI - Growth of lion and puma lentiviruses in domestic cat cells and comparisons with FIV. AB - Feline immunodeficiency virus (FIV-Fca) is a lentivirus that causes gradual immunological deterioration in domestic cats. Lentiviruses related to FIV have been detected in several nondomestic feline species; the biologic significance of these viruses remains to be defined. To examine the in vitro cell tropism of these nondomestic cat lentiviruses, prototypical puma and lion lentiviruses (FIV Pco and FIV-Ple) were cultured in a variety of feline cell cultures. A domestic cat T lymphoma cell line, 3201, best supported the replication of both FIV-Pco and FIV-Ple. Moreover, FIV-Ple was lytic for these cells. RT-PCR amplification of a conserved pol gene region demonstrated species-specific primer homology. Sequence and phylogenetic analyses of this amplification product confirmed the identity of the replicating viruses and classified two previously uncharacterized viruses within predictable lion and puma clades. Sequence analysis of a conserved pol region demonstrated homology with previously characterized FIV-Ple and FIV Pco. Western blot analysis using domestic cat anti-FIV-Fca sera showed that both FIV-Pco and FIV-Ple were antigenically related, to differing degrees, to three serotypes of FIV-Fca. These studies demonstrate that though nondomestic cat lentiviruses differ significantly from FIV-Fca and that a viral-specific protocol may be necessary for sensitive viral detection, these viruses can replicate in cells of domestic cats. suggesting the potential for cross-species transmission. PMID- 9201229 TI - Indicator cell lines for detection of primary strains of human and simian immunodeficiency viruses. AB - CCR5 and CXCR4 are the two major coreceptors that have been identified for human immunodeficiency virus (HIV) entry. We have modified several beta-galactosidase based HIV indicator cell lines to express CCR5 and/or CXCR4. Using these new reagents, we have been able to detect all primary isolates tested using one or both of these cell lines. However, there is large variation in the absolute viral infectivity among primary strains. Furthermore, all HIV strains are capable of causing syncytia in the indicator cells when the coreceptor is present regardless of whether they had previously been characterized as "syncytia-inducing" or "non syncytium-inducing." PMID- 9201230 TI - Coxsackievirus-induced chronic inflammatory myopathy: virus variants distinguish between acute cytopathic effects and pathogenesis of chronic disease. AB - Infection with the Tucson strain of coxsackievirus B1 (CVB1T) causes the development of chronic inflammatory myopathy (CIM) and hind limb weakness in susceptible strains of mice. In this study, a panel of six plaque-purified viruses exhibiting either small or large plaque phenotypes was derived from parental CVB1T and parental CVB1T that had been passaged through monkey kidney cells. All six variants caused similar acute histopathology in muscle, but three of four passaged viruses (AMP1, AMP2, and AMP3) did not induce CIM while the fourth (MP3) caused some hind limb weakness but without associated muscle inflammation. In contrast, both viruses (MP1 and MP2) isolated directly from the parental CVB1T stock were myopathic. Large plaque MP2 caused higher mortality and more rapid inhibition of host cell biosynthesis, but both MP1 and MP2 induced CIM that was comparable to that induced by parental CVB1T. Plaque size was a stable characteristic of the variants but did not correlate with their ability to induce CIM. Five of the six variants showed equivalent levels of replication in muscle, monkey kidney cells, and GB myoblasts while one, AMP3, was selectively impaired for replication. Receptor binding and virus-induced inhibition of host cell transcription and translation were not linked to myopathogenicity. Thus, most of the passaged variants are robust infectious viruses, suggesting that viral induction of CIM does not depend solely on cytopathogenicity during the acute infection. PMID- 9201231 TI - Characterization of P91, a protein associated with virions of an Orgyia pseudotsugata baculovirus. AB - Polyclonal antiserum produced against preoccluded virions from the Orgyia pseudotsugata multinucleocapsid nuclear polyhedrosis virus (OpMNPV) was used to screen an OpMNPV lambda gt11 expression library. The insert from one of the immunoreactive phage isolates hybridized to OpMNPV orf86 (p91), a 2460-bp (819 amino acids) open reading frame that encodes a predicted protein of 91 kDa. Antibodies generated against a maltose binding protein-P91 fusion detected a band of approximately 91 kDa on Western blots of extracts of OpMNPV-infected Lymantria dispar cells. This band was first observed at 18 hr p.i. and was present at all later time points. Similar results using this antiserum were seen with a time course of Autographa californica-infected Spodoptera frugiperda cells. Localization of P91 by confocal immunofluorescence microscopy showed that the protein was concentrated near the nuclear membrane and at late times p.i. was most concentrated near polyhedra. Immunoelectron microscopy indicated that P91 was present in both the capsid and envelope surrounding the capsid of occlusion derived virions. Fractionation studies employing NP-40 and Western blot analysis indicated that P91 was associated with the capsid structure. PMID- 9201232 TI - Avian influenza A viruses differ from human viruses by recognition of sialyloligosaccharides and gangliosides and by a higher conservation of the HA receptor-binding site. AB - Avian influenza virus strains representing most hemagglutinin (HA) subtypes were compared with human influenza A (H1N1,H3N2) and B virus isolates, including those with no history of passaging in embryonated hen's eggs, for their ability to bind free N-acetylneuraminic acid (Neu5Ac) and sialylollgosaccharides in a competitive binding assay and to attach to gangliosides in a solid-phase adsorption assay. The avian viruses, irrespective of their HA subtype, showed a higher affinity for sialyl-3-lactose and the other Neu5Ac2-3Gal-terminated oligosaccharides and a lower affinity for sialyl-6-lactose than for free Neu5Ac, indicative of specific interactions between the HA and the 3-linked Gal and poor accommodation of 6 linked Gal in the avian receptor-binding site (RBS). Human H1 and H3 strains, by contrast, were unable to bind to 3-linked Gal, interacting instead with the asialic portion of sialyl-6-(N-acetyllactosamine). Different parts of this moiety were recognized by H3 and H1 subtype viruses (Gal and GlcNAc, respectively). Comparison of the HA amino acid sequences revealed that residues in positions. 138, 190, 194, 225, 226, and 228 are conserved in the avian RBS, while the human HAs harbor substitutions at these positions. A characteristic feature of avian viruses was their binding to Neu5Ac2-3Gal-containing gangliosides. This property of avian precursor viruses was preserved in early human H3 isolates, but was gradually lost with further circulation of the H3 HA in humans. Consequently, later human H3 isolates, as well as H1 and type B human strains, were unable to bind to short Neu5Ac2-3Gal-terminated gangliosides, an incompatibility that correlated with higher glycosylation of the HA globular head of human viruses. Our results suggest that the RBS is highly conserved among HA subtypes of avian influenza virus, while that of human viruses displays distinctive genotypic and phenotypic variability. PMID- 9201233 TI - U5 region of the human immunodeficiency virus type 1 long terminal repeat contains TRE-like cAMP-responsive elements that bind both AP-1 and CREB/ATF proteins. AB - Activating protein-1 (AP-1) binding phorbol ester responsive elements (TRE) are located downstream of the transcription initiation site in the U5 region of the human immunodeficiency virus type-1 (HIV-1) long terminal repeat (LTR). These downstream sequence elements, termed DSE, can bind cFos and junD and transmit protein kinase C (PKC) activation signals to the LTR. Further studies suggested the DSE might also bind AP-1-related proteins of the CREB/ATF family. Since enhanced HIV-1 expression is associated with activation of the cAMP-dependent protein kinase A (PKA) signaling pathway, we determined whether binding of CREB/ATF proteins to the DSE mediate cAMP/PKA activation of the HIV-1 LTR. In the present study. DSE binding complexes in nuclear protein extracta from colonic epithelial cells are shown to contain ATF-1, ATF-2, and CREB and transfection of either an ATF-2 or PKA expressing plasmid transactivated the DSE. Cholera toxin (Ctx), a potent activator of the cAMP/PKA pathway. Increased HIV-1 virus production from a latently infected promonocytic cell line, U1. Ctx increased LTR promoter activity and increased the CREB content of DSE binding complexes. Transfection of U1 cells with a series of mutant LTR reporter constructs demonstrated that the Ctx response was in large part mediated by the DSE. The Ctx response was also mediated by a heterologous promoter containing multiple TRE sites. Nuclear protein extracts from a T-cell line infected by HIV-1 contained higher levels of CREB/ATF proteins and manifested increased CREB/ATF binding activity. Collectively, these results indicate the DSE are TRE-like cAMP responsive elements that bind both AP-1 and CREB/ATF permitting induction of the HIV-1 LTR by both PKC and PKA activation signals. PMID- 9201234 TI - Control of gene expression during lymphoid development: targeted gene disruption provides new clues. AB - The expression of structural genes is thought to be regulated by DNA-binding factors interacting with cis-acting regulatory elements. These regulatory elements, identified for many lymphopoietic genes, have served in recent years to identify and clone novel transcription factors. The expression of some of these factors is found to be confined to the lymphoid lineage. This regulated expression in both time and space is thought to mediate entry into and progression along the correct developmental differentiation programs. In recent years, many laboratories have tried to assess the functional relevance of these DNA-binding factors by making use of gene targeting techniques. A review of the results of such knock-out experiments and the consequences for lymphoid development models appears below. PMID- 9201235 TI - Gene amplification in the diagnosis of mycobacterial infections. AB - The number of importance of infections caused by Mycobacterium tuberculosis and other mycobacterial species is increasing. Since the detection and identification of mycobacteria by conventional laboratory methods (cultivation, staining, and biochemical tests) is a slow and complex procedure, rapid diagnostic methods are urgently needed. Several amplification methods based on different techniques have been applied in the detection of mycobacteria directly from clinical specimens. Most experience has been obtained from different polymerase chain reaction (PCR) assays and their general performance is good. However, their sensitivity in the analysis of samples containing small amounts of mycobacteria or samples containing inhibitory substances has been low. Furthermore, the risk of false positives caused by contamination is high, and the clinical relevance of the results may be unclear. Thus, these gene amplification techniques are a valuable adjunct to the diagnosis of mycobacteria, but so far they cannot replace conventional microbiological methods. PMID- 9201236 TI - Chronic cold agglutinin disease of the "idiopathic" type is a premalignant or low grade malignant lymphoproliferative disease. AB - We investigated the clinical, pathological, and immunological features of "idiopathic" cold agglutinin disease (CAD) in a population-based study. Fourteen patients were studied, giving a prevalence of about 14 per million with a mean age of 75 years. Haemolysis was present in all cases, but only eight patients had clinical symptoms of peripheral haemagglutination. Serum electrophoresis, immunofixation, morphological bone marrow evaluation, and flow cytometric immunophenotyping were used to detect any monoclonal lymphoproliferative disorder. Flow cytometry seemed to be a sensitive way to demonstrate a clonal B cell proliferation. Some evidence of clonality was found in 13 patients, and a clonal lymphoproliferative disease was documented by flow cytometry or biopsy in 10 out of 11 patients. We conclude that CAD is a symptom-producing monoclonal lymphoproliferative disorder in nearly all patients. PMID- 9201237 TI - Prognosis of radically operated breast carcinoma patients. A retrospective study of 167 consecutive patients with emphasis on histopathological grading, reproducibility and mean nuclear area. AB - A retrospective study of 167 consecutive radically treated breast cancer patients with histopathologically confirmed ductal carcinoma is presented. The aim was to establish the prognostic significance and reproducibility of histopathological grading done independently by two pathologists. Further-more, the value of measurements of mean nuclear area (MNA) in the primary tumour was assessed. The two pathologists reviewed the same histological sections using a three-point scoring system based on tubular structures, number of mitoses and nuclear pleomorphism. Grading was identical for 70% of the tumours (Kappa value 0.51). With increasing MNA, the fraction of poorly differentiated tumours increased. In the univariate analysis, tumour-related survival was significantly related to histopathological grading when G3 tumours were compared to G1/G2 tumours (p < 0.05). In the multivariate analysis, tumour size (pT category), lymph-node status and grading were the only significant factors influencing patient outcome (p < 0.05). MNA had no significant prognostic value. A combination of tumour size and histopathological grading identifies a group of node-negative patients (pT2 G2/G3) who may have a less favourable prognosis and for whom adjuvant treatment may be beneficial. PMID- 9201239 TI - The homogeneity of hidradenitis suppurativa lesions. A histological study of intra-individual variation. AB - Recent data suggest that hidradenitis suppurative is a disease of the follicle, but the histological homogeneity of findings over time and in different anatomical regions has not been verified. Its description may help towards a better classification of the follicular diseases. Correct classification provides both knowledge by interference and a way of generalizing with respect to the significance of specific findings. The intra-individual variation of hidradenitis was described through classification of specimens taken from patients with multiple simultaneous or consecutive excisions. A total of 51 specimens from 11 patients were examined; of these 30 were from synchronous biopsies, and 21 from consecutive biopsies (range 2 months to 6 years). The majority of specimens (44/51) contained poral occlusion, sinus tracts or cysts. This pattern was present in 50-85% of the specimens from any given patient with hidradenitis. No primary apocrine involvement was seen. Fibrosis occurred often (33/51 specimens), and eccrine involvement was seen more often than apocrine involvement (10 vs 7 specimens). The homogeneous histology of hidradenitis supports its reclassification as a follicular disease. The reproducibility of the findings further suggests that the changes are specific and that hidradenitis is therefore a definite disease entity within the spectrum of follicular diseases. Additional functional studies may help classify hidradenitis more precisely in relation to other follicular diseases. PMID- 9201238 TI - Serum concentration of the aminopropeptide of type I procollagen in patients on haemo- and peritoneal dialysis. AB - Using an ELISA technique, the aminopropeptide of type I procollagen (PINP) was measured in serum from patients with chronic renal failure treated with haemodialysis (HD) (n = 19) or continuous ambulatory peritoneal dialysis (CAPD) (n = 14), and compared to the commonly used bone markers. The serum concentrations for PINP, compared to healthy controls were significantly increased in both the HD-group (p < 0.00001) and the CAPD-group (p < 0.00001). In the HD-group a close correlation was found between PINP and parathyroid hormone (PTH) (R(s) = 0.745; p = 0.00026) and between PINP and total alkaline phosphatase (R(s) = 0.623; p = 0.004), but in the CAPD-group the corresponding p-values were 0.17 and 0.06 only. No significant difference was found between the HD and CAPD patients with respect to serum levels of PINP, PTH, total alkaline phosphatase, or ionized calcium. In the HD-patients, a significantly higher level of serum phosphate was found compared to in the CAPD-patients. The present study demonstrates a close correlation between PTH, total alkaline phosphatase and PINP, which indicates that PINP might be used as a marker for evaluating increased bone turnover in patients with chronic renal failure treated with haemodialysis, and perhaps also in patients treated with peritoneal dialysis, and that the ideal biochemical parameters to analyse changes in bone metabolism in these patients may be a combination of the initiating hormone (PTH) and PINP as a marker of the effect of PTH on bone metabolism. PMID- 9201240 TI - Expression of the ubiquitin carboxyl-terminal hydrolase PGP 9.5 in axons following spinal cord compression trauma. An immunohistochemical study in the rat. AB - Protein gene product 9.5 (PGP 9.5) is a neuron-specific protein which acts as a ubiquitin carboxyl-terminal hydrolase. It facilitates the conversion of polyubiquitin to monoubiquitin, which can be reused for another catalytic cycle. Monoubiquitin plays an important role in degrading abnormal and denatured proteins. Previously, we have reported that ubiquitin-like immunoreactivity is expressed in axonal swellings following compression trauma to the rat thoracic cord. It was characterized by fast occurrence, progressive increase and gradual disappearance over a period of 9 days. The expression of PGP 9.5 has now been studied in the same material. Control rats showed a weak PGP 9.5 immunoreactivity in the nerve cell bodies of the cord. Except for the corticospinal tracts, the axons of other longitudinal tracts were weakly stained. Accumulation of PGP 9.5 immunoreactivity occurred in expanded axons at the site of compression already 4 h after trauma. They became more frequent in number 1 and 4 days after injury and remained so over the entire observation period of 9 days. The labelled axons were randomly distributed in the longitudinal tracts, but were never found in the corticospinal tracts. The extent of immunoreactivity was related to the degree of impact on the cord. Compression injury thus induces accumulation of both ubiquitin and PGP 9.5 immunoreactivity in axonal expansions. The injured axons may have a mechanism for degradation of proteins by the ubiquitin-mediated proteolytic pathway and another mechanism for effective ubiquitin regenerative cycling by the action of PGP 9.5. PMID- 9201241 TI - Mechanisms involved in the early interaction between HeLa cells, platelets and endothelial cells in vitro under the influence of thrombin. Effects of acetylsalicylic acid and Na-salicylate. AB - The aim of the study was to obtain more information about the mechanisms involved in the initial adhesion of tumour cells to endothelial cell during metastasis. In a previous paper, we found that addition of both platelets and thrombin increased the adhesion of tumour cells to cultured endothelial cells within 15 min, compared to when either one or both of the ingredients were absent. In the present study, HeLa cells, prelabelled with radioactive 51Cr, human platelets, and thrombin, were added to the medium in dishes of endothelial cells. The dishes were then shaken for 15 min at 37 degrees C. Scanning and transmission electron micrographs showed HeLa cells adhering to the endothelium either together with platelets or without them. In other experiments, the endothelium was pretreated for 30 min with either of the following: 0.5 mM or 0.1 mM acetylsalicylic acid (ASA); 0.5 mM or 0.1 mM Na-salicylate (NaS). Pretreatment of the endothelium with 0.5 mM ASA significantly increased the percentage of adherent tumour cells, while 0.1 mM ASA and the two concentrations of NaS caused only minor changes. In addition, the ASA-treatment caused more HeLa cells to adhere without platelets while NaS-treatment caused more HeLa cells to adhere together with platelets. Release of 51Cr from HeLa cells during the experimental period was also measured; the addition of thrombin and platelets did not change the 51Cr release significantly. In separate experiments, HeLa cells and platelets were mixed without the presence of endothelial cells. Transmission electron micrographs showed that in the absence of thrombin, mixed HeLa cells and platelets did not react with each other; when thrombin was added they formed co-aggregates. In conclusion, we show that in our experimental model HeLa cells adhere to the endothelium in two ways, both with and without platelets. The production of prostacyclin in the endothelial cells has an inhibitory effect on tumour cell adhesion. Without thrombin, the HeLa cells are not capable of activating platelets. PMID- 9201242 TI - Effect of adenovirus 2 on cellular gene activation in blood-derived monocytes and macrophages. AB - We have investigated the effect of adenovirus 2 (Ad2) infection on human monocytes and monocyte-derived macrophages with regard to expression of TNF-alpha and IL-1 beta. In monocytes, the virus was bound to the surface without being internalized. On the other hand, Ad2 was internalized by macrophages. No virus replication and no transcription of the Ad2 early genes was observed in either of the cells. Ad2 infection induced transient increase in the mRNA levels for TNF alpha and IL-1 beta in both monocytes and in macrophages, although the kinetics of the transcription was slightly different. The production of both cytokines, measured by ELISA tests, was enhanced in monocytes. In macrophages, a slight enhancement of TNF-alpha production was seen, whereas IL-1 beta was not detected. The data indicate that cellular genes might be activated by Ad2 virus infection in nonpermissive cells where no viral gene products could be detected. PMID- 9201243 TI - Transfer of primed CD4+OX40- T lymphocytes induces increased immunity to experimental Salmonella typhimurium infections in rats. AB - The protective effect of primed CD4 T cells against a lethal dose of Salmonella typhimurium was studied in Lewis rats. Primed CD4 T cells were obtained by inoculating Lewis rats with a non-lethal dose of S. typhimurium. Four weeks after the infection, spleen non-adherent mononuclear cells were isolated. The cells were separated according to their expression of CD4 and the OX40 antigen by FACS. OX40+ and OX40- CD4+ T-cell subpopulations were together with unsorted CD4+ T cells transferred to untreated rats 24 h prior to infection with S. typhimurium. Transfer of either unsorted CD4+ T cells or CD4+ T cells sorted into OX40- or OX40- subpopulations significantly increased animal survival compared to controls. Animals receiving OX40+CD4+ T cells did not differ significantly in survival probability from those receiving unsorted CD+ T cells. However, animals receiving OX40-CD4+ T cells had a significantly better survival compared to animals given unsorted CD4+ T cells. It is concluded that OX40-CD4+ T cells can induce significant protection against S. typhimurium infections in rats. This is most likely due to the fact that the OX40-CD4+ T-cell population contains a significant number of antigen-specific memory T cells that have returned to a resting state. PMID- 9201244 TI - Sclerosing peritonitis in a case of benign cystic ovarian teratoma. A case report. AB - A 52-year old woman with regular menstruation presented with ascites and abdominal swelling and pain. Bilateral salpingo-oophorectomy, hysterectomy and excision of a few peritoneal nodules was performed. The omentum was firm, giving the impression of carcinoma. The ovaries were enlarged, each containing a non ruptured cyst with thick yellowish fluid. Microscopy revealed a sclerosing peritonitis in the omentum and nodules. There was a benign cystic teratoma in the left ovary, a corpus luteum in the right ovary. Follow-up has been uneventful for 26 months. The sclerosing peritonitis is considered to be secondary to the ovarian changes, most probably the teratoma. PMID- 9201245 TI - Clinical response versus clinical benefit in oncology: not necessarily equivalent terms. PMID- 9201246 TI - Regulation of the release of tumour necrosis factor (TNF)alpha and soluble TNF receptor by gamma irradiation and interferon gamma in Ewing's sarcoma/peripheral primitive neuroectodermal tumour cells. AB - This study analyses the production of tumour necrosis factor (TNF)alpha and soluble TNF receptor (sTNF-R) before and after exposure to gamma irradiation and interferon gamma (IFN gamma) in 12 cell lines derived from Ewing's sarcoma (ES)/peripheral primitive neuroectodermal tumours (pPNET). Supernatants from ES/pPNET cell cultures were tested in a TNF alpha-specific amplified enzyme linked immunosorbent assay (ELISA), a bioassay, and sTNF-Rp55 and sTNF-Rp75 ELISA. The tumour cell lines released minimal amounts of TNF alpha, prominent amounts of sTNF-Rp55 (7/12 cell lines) and no sTNF-Rp75. Exposure to gamma irradiation (5 Gy) either induced (3/12) cell lines) or up-regulated (3/12 cell lines) TNF alpha release without changing sTNF-Rp55 and sTNF-Rp75 levels. Priming of cultures with recombinant human IFN gamma (rhIFN gamma) markedly enhanced TNF alpha secretion in the radiation-responsive cell lines and had no influence on sTNF-Rp55 and sTNF-Rp75 levels. rhIFN gamma affected the magnitude rather than the sensitivity of the radiation response. The TNF alpha secreted was bioactive, as shown by its cytotoxic effect of WEHI-164 cells, and neutralization of its activity by anti-TNF alpha monoclonal antibody. Herbimycin A (a tyrosine-specific protein kinase inhibitor) but not calphostin C (a protein kinase C inhibitor), H89 (a protein kinase A inhibitor), AA-COCF3 (a specific inhibitor of phospholipase A2) and MK-886 (a specific inhibitor of 5-lipoxygenase) abrogated gamma-irradiation-stimulated TNF alpha release. The antioxidants N acetylcysteine, nordihydroguaiaretic acid and mepacrine dose-dependently inhibited gamma-irradiation-mediated TNF alpha production. Collectively our findings indicate that IFN gamma priming potentiates the secretion of bioactive TNF alpha by ES/pPNET cells in response to gamma irradiation without affecting sTNF-R release. The data suggest a requirement for protein tyrosine kinase activity and a role for reactive oxygen species in the gamma-irradiation-mediated intracellular signalling pathway leading to TNF alpha production. PMID- 9201247 TI - Genotoxic changes in the pulmonary alveolar macrophages of mice, rats and hamsters treated with tobacco smoke. AB - To determine whether tobacco smoke (TS) is genotoxic for lung tissue macrophages (pulmonary alveolar macrophages, PAM) as a general result of its inhalatory action BD6 rats, Syrian golden hamsters and BDF1 (C57BlxDBA2) mice were subjected to wholebody exposure for 90 or 60 min daily (600 cm3 mainstream smoke in 16-1 glass chamber, 9 or 6 exposures of 15 min each, respectively), for different periods ranging up to 30 days. A significant enhancement of the frequency of polynucleated macrophages (BiN PAM) was observed in all animal species after more than 10-days of repeated exposure to TS. The increased level of BiN PAM (the number of bi- (+) poly-nucleated PAM) correlates with the duration of exposure to TS: on day 20 after the start of inhalation, more than 25/1000 of mice PAM were polynucleated, while on day 30 this applied to approximately 50/1000. Furthermore, a highly significant increase in the level of micronucleated PAM (MN PAM) was also established after 10 days TS treatment of mice and persisted to the end of these examinations. TS was effective in enhancing the micronucleated and polynucleated PAM levels in hamsters irrespective of their sex, as it was in male BD6 rats aged 2 or 11 months. It appears that TS induces a more pronounced elevation of polynucleated PAM frequency in rats than in hamsters and mice. These data suggest that inhaled TS is genotoxic in alveolar macrophages in all exposed species of laboratory animals. An attempt was made to trace the possible clastogenic effect of a single i.p. administration of cyclophosphamide (CP, 15 mg/kg) in mice simultaneously in bone marrow and in PAM. A definite clastogenic effect in bone marrow 24 h and 48 h after CP injection and a total absence of changes in PAM from the lungs during the 15-day period after clastogen exposure were established. These data may support the hypothesis of local production of PAM in the lung from their proliferative precursor. The results provide evidence that PAM in laboratory animals are a sensitive and useful target for assessing harmful effects associated with environmental chemical factors that can be inhaled, including TS. PMID- 9201248 TI - A practical prognostic index for inoperable non-small-cell lung cancer. AB - Radical radiotherapy is widely used to treat inoperable non-small-cell lung cancer (NSCLC) although only a small number of patients benefit in the long run from this intensive treatment. There is a small proportion of long-term survivors who might derive advantage from even more aggressive radiotherapy combined with chemotherapy. In order to support optimal treatment selection we have carried out univariate and multivariate analyses of possible prognostic variables in the retrospective data of 502 NSCLC patients treated at one institute with external radiotherapy, both with curative and palliative intent. To obtain more accurate tools for a rational treatment decision, we identified, by using Cox's proportional-hazards model, the five most powerful determinants of overall survival and combined them to a prognostic index. On the basis of only the number of these risk factors (advanced stage, general or metastatic symptoms, poor performance status, anemia and tumor size of at least 7 cm), the patient falls into one of the six possible prognostic groups and these groups turned out to be identifiable as separate prognostic clusters. Thirty-one per cent of the patients have three or more risk factors and a median survival of 5-7 months compared with 18 months for patients without any non-favorable factor. Furthermore, the prognostic factors were so strong that multivariate analyses did not reveal the treatment selection to have any significant influence on survival. As each of the five variables have the advantage of being routinely available, our index is simple enough to be used in daily clinical practice. The clinical value of the prognostic index should be verified by using independent data. PMID- 9201249 TI - Effect of the cytostatic butyric acid pro-drug, pivaloyloxymethyl butyrate, on the tumorigenicity of cancer cells. AB - Previously we have shown that pivaloyloxymethyl butyrate (AN-9), a pro-drug of butyric acid (BA), is a differentiation-inducing agent in a variety of cells. In this report, we demonstrate that AN-9 is a cytostatic but not cytotoxic agent in a myelomonocytic cell line (WEHI); thus, the cells were growth-arrested and differentiated. These late changes in the cells were preceded by changes in the expression of the early regulatory genes, c-myc and c-jun. Although initiation of all these events had already occurred after 1 h exposure to AN-9, the tumorigenicity of these cells tested in Balb/c mice was not affected. A marked reduction in the tumorigenicity of AN-9-treated cells was observed after 4 h of exposure. Exposure of the highly metastatic subclone of Lewis lung carcinoma (3LLD122) to AN-9 resulted in a very pronounced effect on the tumorigenicity of these cells tested in C57BL mice. Unlike WEHI cells, the tumorigenicity of 3LLD122 was almost completely diminished after 1 h of exposure. In both cell types a 10-fold higher concentration of BA did not affect the tumorigenicity of the cells as did AN-9. PMID- 9201250 TI - Modifying effect of reproductive risk factors on the age at onset of breast cancer for German BRCA1 mutation carriers. AB - Female carriers of mutations in the BRCA1 gene on chromosome 17q have a very high risk of developing breast and/or ovarian cancer during their lifetime. There is, however, little knowledge of to what extent non-genetic risk factors, such as age at menarche, age at first birth, and body mass index, alter the age at onset of disease. We identified individuals showing a high probability of linkage to BRCA1 and examined the effect of other known risk factors on disease risk. A total of 43 families with at least three breast or ovarian cancer cases, including two affected before 60 years of age, were studied for linkage to the susceptibility locus BRCA1. Blood samples from relevant family members were used to genotype for at least three chromosome 17q polymorphic markers. Information on reproductive history, hormone use and lifestyle factors was collected from female members using a self-administered questionnaire. Diagnoses of breast and ovarian cancer were verified through pathology reports and paraffin blocks were obtained when available. Multipoint LOD (logarithm of the odds) scores were calculated and individuals from 10 families with a posteriori probability for linkage greater than 0.90 were used for further analysis. Forty-six BRCA1 carriers were identified by the disease haplotype; 30 were affected with breast cancer and 5 with ovarian cancer. Proportional- hazards analysis of age at onset of breast cancer yielded increased relative risks of 1.74 for early age at menarche (< 14 years), 1.58 for late age at first birth (> or = 25 years) or nulliparity, and 2.78 for recent year of birth (> or = 1940); however, none of the risk estimates was statistically significant. When both breast and ovarian cancer were considered as disease endpoints, the birth cohort effect was stronger and age at first birth showed no effect. Our data provide some evidence that reproductive risk factors for breast cancer have an effect on age at onset for BRCA1 carriers. However, considering that our analyses were based on limited numbers, these results warrant further clarification. PMID- 9201253 TI - Interdisciplinary forum: from Genetic Diagnosis to Gene Therapy in Oncology. 26 September-1 October, 1996, Madrid, Spain. PMID- 9201252 TI - The development of alkylphosphocholines as signal transduction inhibitors: experimental and clinical challenges. AB - Alkylphosphocholines are a new class of anticancer agents. Their mode of action is considered to be related to the inhibition of phospholipase C and protein kinase C. These enzymes play a major role in intracellular signalling pathways. Their inhibition by alkylphosphocholines leads in the dimethylbenzanthracene induced mammary carcinoma of the rat to a response pattern similar to that of the antiestrogen zindoxifene. This suggests that the inhibition of transcription factor formation might be the common pathway for alkylphosphocholines and antihormones. Based on the experimental dose-response pattern, new clinical strategies for dose finding and response evaluation will have to be developed for inhibitors of signal transduction, such as alkylphosphocholines. PMID- 9201251 TI - Increased phospholipase D activity in human breast cancer. AB - Phospholipase D is believed to play an important role in cell proliferation and tumorigenesis. One of its major functions is to cause a sustained activation of protein kinase C through the primary production of phosphatidic acid from phosphatidylcholine by the enzyme, followed by dephosphorylation forming diacylglycerol. Protein kinase C is known to be activated or translocated in some tumors including breast tumors. In order to examine phospholipase D activity in breast tumors, surgical specimens of human breast tumors were obtained by mastectomy or wide excision, and their phospholipase D activities were assayed by determining the formation of phosphatidylethanol from phosphatidylcholine and ethanol. Phospholipase D activity was predominantly localized in the microsomal fraction of the tumor tissue and markedly stimulated by oleic acid. We observed a significant increase in phospholipase D activity in 17 out of 19 spontaneous human breast tumors as compared to adjacent histologically normal breast tissue. The mean specific activity in the tumors was 52.9 +/- 41.8 (SD) pmol min-1 mg protein-1 whereas the value for the normal breast tissue was 34.0 +/- 36.2 (SD) pmol min-1 mg protein-1 (P < 0.01; paired Wilcoxon's rank-sum test). The mean tumor/normal activity ratio was 2.37. Among prognostic factors, the nuclear grade, evaluated according to Schnitt et al., was found to be correlated with the activity ratio. Our results suggest a role for phospholipase D in human breast tumors. An elevation in phospholipase D activity is useful as a potential marker for malignant disease in the breast. PMID- 9201254 TI - Changes in N-acetylglucosaminyltransferase III, IV and V in renal cell carcinoma. AB - The activities of N-acetylglucosaminyltransferase (GnT) III, IV and V were determined in 10 cases of renal cell carcinoma (RCC) and compared with the normal kidney cortex (NKC) regions of the same kidney resected from RCC patients. It was found that the GnT III and GnT IV activities decreased consistently in all samples of RCC, while GnT V activity increased, decreased or did not change in different samples. The mean levels of GnT III and GnT IV activities in RCC were found to be very significantly lower than those of NKC on statistical analysis, but the mean value of GnT V activity was almost identical in RCC and NKC. The decrease in GnT activities in RCC were compatible with the decrease in bisecting N-acetylglucosamine (GlcNAc) and antennary number of complex-type N-glycans in gamma-glutamyltranspeptidase (gamma-GT) partially purified from RCCs as studied with concanavalin A (ConA) affinity column chromatography, which showed a decrease of unbound fraction and increase of bound fractions. PMID- 9201255 TI - Mechanisms of neutrophil-induced parenchymal cell injury. AB - Neutrophils are involved in organ damage induced by an excessive acute inflammatory response after ischemia-reperfusion, trauma, and sepsis. In addition to causing vascular injury, neutrophils can transmigrate and attack parenchymal cells. This review summarizes recent advances in our understanding of neutrophil induced parenchymal cell injury using the liver as an example. Reviewed are the mechanisms of neutrophil sequestration in the hepatic vasculature, transendothelial migration, adherence to hepatic parenchymal cells, and mechanisms of cytotoxicity. Discussed are the involvement of various adhesion molecules in these processes, the role of cytokines and chemokines in the pathophysiology, as well as the effects of proteases and reactive oxygen species released by neutrophils. The emerging understanding of the basic mechanisms of neutrophil-induced organ damage is critical for the development of therapeutic strategies to attenuate excessive acute inflammatory responses without compromising essential host defense mechanisms. PMID- 9201257 TI - Expression of TNF-alpha by human plasma cells in chronic inflammation. AB - Tumor necrosis factor-alpha (TNF-alpha) is a potent pro-inflammatory cytokine and mediator of the inflammatory response. It has been implicated in the pathogenesis of many inflammatory disorders, including rheumatoid arthritis (RA), septic shock, and Crohn's disease. Using a specific anti-human TNF-alpha antibody we detected immunoreactivity for this cytokine in the cytoplasm of inflammatory cells in several chronic inflammatory disorders, including RA, scleritis, and polyarteritis nodosa. These cells were identified predominantly as IgG-expressing plasma cells. Lymph nodes from patients with Hodgkin's lymphoma and breast cancer, but not from control subjects, were also found to contain TNF-alpha positive plasma cells. Cultured EBV-B lymphocytes and a human plasma cell line (ARH-77) when stimulated with phorbol myristate acetate demonstrated cytoplasmic TNF-alpha immunoreactivity. Western blot analysis of cell membranes and conditioned media from both cell types revealed the presence of the 26-kDa membrane-bound from and the 17-kDa soluble from of TNF-alpha, respectively. TNF alpha was quantitated by enzyme-linked immunosorbent assay and found to be biologically active as determined by the L929 cytotoxicity assay. This is the first demonstration that plasma cells may be capable of modulating immune and inflammatory responses, not only by antibody production, but also by their secretion of a key inflammatory mediator, TNF-alpha. PMID- 9201256 TI - Immunosuppressive retroviral peptides: immunopathological implications for immunosuppressive influences of retroviral infections. AB - Studies of the effects of retroviruses on the immune system, which date back through thirty years of investigations, are reviewed. In the earliest published studies in the 1960s, it was demonstrated that mice infected with oncogenic viruses were immunosuppressed. Since then, numerous articles have been published describing profound immunodeficiencies observed in vivo in humans infected with human immunodeficiency virus and in animals such as cats infected with the feline immunodeficiency virus. In vitro investigations have shown that inactivated retroviruses or transmembrane envelope protein p15E as well as a synthetic 17 amino acid peptide (CKS-17) impressively conserved within the transmembrane envelope protein of several animal or human retroviruses are highly immunosuppressive. More recently, dysfunction of cytokines produced by CKS-17 at both a cellular and molecular level have been found to mimic influences observed in vivo in patients infected with the human immunodeficiency virus. CKS-17 has also been shown to induce cAMP in vitro. The significance of these observations to understanding the immunological disturbances observed in malignancy, cytokine biosynthesis, and modulations of immune functions through cAMP is discussed. PMID- 9201258 TI - Monocytic-endothelial cell interaction: regulation of prostanoid synthesis in human coculture. AB - Coculture of a monocytic cell line (HL-60) and iliacal endothelial cells as an in vitro model of vascular inflammation was investigated for cooperative regulatory mechanisms of prostanoid synthesis under conditions of selective prestimulation. In coculture of endothelial cells and 12-O-tetradecanoylphorbol 13-acetate (TPA) prestimulated monocytic HL-60 cells the capacity of prostanoid synthesis from arachidonic acid was strongly increased compared with monocultures. Concomitant with up-regulation of specific adhesion molecules, cyclooxygenase (COX) 2 mRNA was induced in endothelial cells in coculture independent of cell contact. HL-60 cells exhibited no alterations in mRNA expression of cyclooxygenases or thromboxane synthase. Coculture of TPA-prestimulated endothelial cells with unstimulated HL-60 cells led to a selectively increased capacity of thromboxane production. Under this condition HL-60 cells up-regulated COX1 and COX2 mRNA, whereas endothelial mRNA levels did not change. Our data demonstrate that the increase in prostanoid synthesis in coculture essentially depends on rapid induction of COX2 mRNA within 2 h. PMID- 9201259 TI - LPS-induced blood neutrophilia is inhibited by alpha 1-adrenoceptor antagonists: a role for catecholamines. AB - A role for catecholamines in the regulation of the blood neutrophilia induced by intravenous (i.v.) injection of lipopolysaccharide (LPS; 250 micrograms/kg) was examined in Wistar rats by means of surgical adrenalectomy or pretreatment with adrenergic and dopaminergic antagonists into naive animals. Treatment of animals with a single dose (250 micrograms/kg) of LPS caused a dramatic increase in the number of circulating neutrophils concomitant with a decrease in the number of these cells in the bone marrow. These effects were partially reversed when catecholamine stores were depleted with reserpine. It was found that neither adrenalectomy nor pretreatment with the dopaminergic antagonists, chlorpromazine and pimozide, affected the changes in neutrophil counts induced by LPS. The injection of the alpha 1/alpha 2-adrenoceptor antagonist, phentolamine, and the selective alpha 1-adrenoceptor antagonist, prazosin, significantly decreased blood neutrophilia induced by LPS. However, neither the selective alpha 2 adrenoceptor antagonist, yohimbine, nor the beta-adrenoceptor antagonist, propranolol, had any effect on LPS response. Taken together, these findings support the hypothesis that the catecholamine norepinephrine plays a role in the regulation of the LPS-induced neutrophilia through activation of alpha 1 adrenoceptors. PMID- 9201260 TI - Secretory leukocyte proteinase inhibitor is a major leukocyte elastase inhibitor in human neutrophils. AB - Secretory leukocyte proteinase inhibitor (SLPI) is the main neutrophil elastase (HLE) inhibitor found in the upper airways during pulmonary inflammation. It has been shown to be synthesized and secreted in vitro by epithelial cells and has been localized in tracheal glands and bronchiolar epithelial cells by immunocytochemistry. In this study, using immunodetection and immunopurification techniques with specific anti-SLPI immunoglobulin G (IgG), we show that SLPI is present as a native 14-kDa molecule in neutrophil cytosol. In addition, we demonstrate that SLPI is the major inhibitor of HLE present in neutrophil cytosol because pre-incubation with specific anti-SLPI IgG was able to inhibit completely the anti-HLE activity of the cytosol. SLPI can be secreted (probably in an inactive form) by neutrophils and its secretion is enhanced when the cells are stimulated with phorbol myristate acetate (PMA). Elafin, an elastase-specific inhibitor, is also present in minute amounts in neutrophil cytosol and its secretion can be up-regulated. The presence of SLPI in the cytosol of neutrophils may serve as a protective screen against proteinases spilling from azurophilic granules. An alternative or supplementary role may be the maintenance of a differentiated phenotype. PMID- 9201261 TI - Complex regulation of human neutrophil activation by actin filaments: dihydrocytochalasin B and botulinum C2 toxin uncover the existence of multiple cation entry pathways. AB - In human neutrophils, the chemotactic peptide, N-formyl-L-methionyl-L-leucyl-L phenalalanine (fMLP), the Ca(2+)-ATPase inhibitor, thapsigargin, and the lectins, concanavalin A (Con A) and mistletoe lectin I (ML I), stimulate the entry of Ca2+ and Na+ with subsequent activation of exocytosis and superoxide anion (O2-) formation. We studied the role of actin in neutrophil activation. The actin filament-disrupting substances, dihydrocytochalasin B (dhCB) and botulinum C2 toxin (C2 toxin) potentiated fMLP- and lectin-stimulated Ca(2+)- and Na+ entry. Lectin-induced Mn2+ entry was enhanced by actin disruption, whereas fMLP triggered Mn2+ entry was unaffected. dhCB and C2 toxin inhibited fMLP- and lectin stimulated Ba2+ influx. The actin disrupters also inhibited fMLP- and ML I induced Sr2+ influx, whereas Con A-stimulated Sr2+ entry was not influenced by dhCB and C2 toxin. Thapsigargin-stimulated cation entry was not altered by actin disruption. DhCB and botulinum C2 toxin potentiated lysozyme release induced by all four stimuli. Con A and ML I per se activated O2- formation only in the presence and not in the absence of dhCB. Con A potentiated the stimulatory effects of ML I on O2- formation in the presence of dhCB and primed neutrophils to respond to ML I in the absence of dhCB. Our data indicate the following: (1) dhCB and C2 toxin uncover the existence of multiple cation entry pathways in neutrophils; (2) actin disruption facilitates exocytosis and O2- formation by enhancement of Ca(2+)- and Na+ entry and by altering the function of proteins involved in activation of secretion and O2- formation; and (3) Con A and ML I, which possess different sugar specificities, activate different signaling pathways in neutrophils. PMID- 9201262 TI - Timing of prostaglandin exposure is critical for the inhibition of LPS- or IFN gamma-induced macrophage NO synthesis by PGE2. AB - Macrophage nitric oxide (NO) synthesis is an integral component of the host defense system. We have previously found that NO and prostaglandins interact in a variety of ways. NO modulates Kupffer cell prostaglandin E2 (PGE2) production and we have recently described the inhibitory effects of PGE2 on NO synthesis in both Kupffer cells and hepatocytes. Activated macrophages produce a number of prostaglandins but studies regarding the capacity of prostaglandins to regulate macrophage NO synthesis have yielded conflicting results. We found that exogenous PGE2 decreased lipopolysaccharide (LPS)-induced NO synthesis in murine resident peritoneal macrophages and in the RAW 264.7 murine macrophage cell line. PGE2 also suppressed NO synthesis in response to interferon-gamma (IFN-gamma) alone and a combination of LPS + IFN-gamma. Inhibition of endogenous PGE2 synthesis with indomethacin or ibuprofen had no effect on NO synthesis. PGE2 added with the activating stimulus was most effective. PGE2 lost the capacity to block NO synthesis if added more than 180 min after LPS. PGE2 decreased inducible NO synthesis (iNOS) mRNA and immunoreactive iNOS protein, consistent with the hypothesis that exogenous PGE2 inhibits macrophage iNOS expression but that the inhibition depends on the time and concentration of prostaglandin exposure. PMID- 9201263 TI - Quantitation of surface CD14 on human monocytes and neutrophils. AB - The absolute number of membrane-expressed CD14, the most important endotoxin receptor, on human monocytes and neutrophils shows remarkable variation in the literature. To quantify these numbers two fluorescence methods using fluorescein isothiocyanate (FITC)-labeled monoclonal antibodies (mAb) were applied. A commercially available set of standard beads was used in flow cytometry to quantitate CD14 with eight different mAbs. Independent from their isotype the various mAbs showed minor differences and indicated that peripheral blood monocytes expressed 99,500-134,600 (115,400 +/- 10,600) and neutrophils 1,900 4,400 (3,300 +/- 800) CD14 receptors. There was no significant difference in CD14 expression on leukocytes in unprocessed freshly obtained whole blood and after a Ficoll isolation procedure. However, a short temperature shift resulted in a 1.3- to 1.6-fold up-regulation of CD14. The results obtained with the reference beads were verified with fluorescence Scatchard analysis and spectrofluorometry using mAb 26ic-FITC and showed 109,500 CD14 per monocyte and 6,700 CD14 per neutrophil. For comparison the number of CD14 on the monocytic THP-1 cells and Fc gamma receptors on human leukocytes were determined using the reference beads and flow cytometry and gave results comparable to published data. Our data indicate that resting human monocytes express roughly 110,000 CD14 molecules on their surface using a simple fluorometric assay. Correct determination of the number of CD14 and other cell surface receptors is of importance in the monitoring of septic patients. PMID- 9201264 TI - IL-15 is chemotactic for natural killer cells and stimulates their adhesion to vascular endothelium. AB - Interleukin-15 (IL-15) is a recently described cytokine with IL-2-like stimulating activities on T lymphocytes and natural killer (NK) cells. IL-15 mediates its function through the beta- and gamma-chains of the IL-2 receptor. In this work, we have investigated the effect of IL-15 on the directional migration of NK cells in chemotaxis assays and on the ability of NK cells to bind to vascular endothelium. IL-15 (10-20 ng/mL) had chemotactic effects on freshly isolated resting NK cells as well as on long-termed IL-2-cultured NK cells. A checkerboard experiment demonstrated that migration in response to IL-15 was observed only in the presence of a positive gradient (chemotaxis). Overnight treatment of freshly isolated NK cells with IL-15 (10-20 ng/mL) augmented their binding to cultured endothelial cells (EC) in vitro, especially to resting EC. IL 15-activated NK cells bound to resting and tumor necrosis factor-activated EC by use of LFA-1/ICAM-1 and VLA-4/VCAM-1 adhesion pathways, essentially as untreated NK cells do. The fact that IL-15 increased NK cell binding to ICAM-1-transfected NIH-3T3 fibroblasts, together with the finding that IL-15 did not increase binding to extracellular matrix proteins, where the major molecules involved are VLA proteins, indicated that IL-15 primarily stimulates LFA-1-dependent adhesion. By increasing NK cell adhesion to vascular endothelium and migratory response, IL 15 is an important determinant of NK cell recruitment in tissues. PMID- 9201265 TI - Primary structure of rat CD14 and characteristics of rat CD14, cytokine, and NO synthase mRNA expression in mononuclear phagocyte system cells in response to LPS. AB - Rat CD14 cDNA clones were isolated. The predicted protein sequence exhibits 82, 61.6, and 64% identity with the mouse, rabbit, and human CD14, respectively. The levels of rat CD14 mRNA expression in resident peritoneal macrophages (PM), alveolar macrophages (AM), and peripheral blood monocytes (BM) were constitutively high, whereas that in Kupffer cells (KC) was low. On intravenous injection with lipopolysaccharide (LPS), the expression of rat CD14 mRNA in KC increased markedly, whereas the increases in PM, AM, and BM were mild. Similar features of expression of rat CD14 in these cells were observed after stimulation with LPS in vitro. The level of tumor necrosis factor alpha (TNF-alpha) mRNA expression in KC after stimulation with LPS in vivo was comparable to that in PM, AM, and BM, whereas that of TNF-alpha mRNA expression in KC and PM after stimulation with LPS in vitro was lower than that in AM and BM. Interleukin (IL) 1 beta and iNOS mRNA expressions in KC after stimulation with LPS in vivo and in vitro were low, whereas those in PM, AM, and BM were high. Little or no expression of IL-6 was observed in KC after stimulation with LPS in vivo and in vitro, whereas higher expression was observed in PM, AM, and BM than in KC. PMID- 9201267 TI - Desensitization of the fMLP-induced NADPH-oxidase response in human neutrophils is lacking in okadaic acid-treated cells. AB - The chemoattractant N-formyl-methionyl-leucyl-phenylalanine (fMLP) interacts with neutrophils, generating signals that induce activation of the superoxide anion/hydrogen peroxide-producing NADPH-oxidase. Low temperature binding of fMLP to its neutrophil surface receptors is associated with a desensitization of the cells with respect to activation of the oxidase. Other stimuli can still activate the oxidase (in fact even induce a primed response), indicating that the observed phenomenon is stimulus specific and could not be accounted for by an effect on the oxidase itself. Furthermore, no desensitization is obtained in the presence of cytochalasin B, suggesting that the cytoskeleton is involved in the process leading to desensitization. Okadaic acid is a toxin produced by dinoflagellates and exerts its effects by an inhibition of cellular phosphatases. To investigate the role of phosphorylation/dephosphorylation events in the desensitization process we used okadaic acid as a scientific tool. We show that neutrophils treated with okadaic acid are primed with respect to the fMLP-induced production of superoxide anion, and that no desensitization is obtained in toxin-treated cells. Because the recovery of ligand-receptor complexes in a Triton X-100 insoluble fraction is very low in the cells treated with okadaic acid, we suggest that protein dephosphorylation is required to obtain binding to the cytoskeleton of occupied fMLP receptors; binding of the occupied receptors to the cell cytoskeleton being the mechanism behind desensitization. PMID- 9201266 TI - Inhibitory effect of 3,4-dichloro-propionaniline on cytokine production by macrophages is associated with LPS-mediated signal transduction. AB - Our previous studies demonstrated that both in vivo and in vitro 3,4-dichloro propionanilide (propanil) exposure inhibited interleukin-6 (IL-6) and tumor necrosis factor (TNF) production by adherent thioglycollate-elicited peritoneal cells (macrophages) after lipopolysaccharide (LPS) stimulation. In this study, we report that IL-6 and TNF-alpha message is reduced by propanil in a concentration dependent pattern, yet the stability of cytokine mRNA is not affected. In addition, exposure of macrophages to propanil after a relatively short period of LPS stimulation significantly reduced the production of IL-6 and TNF. Determination of the intracellular Ca2+ levels demonstrates that LPS-induced Ca2+ release is abrogated in propanil-treated macrophages. However, the binding of LPS to macrophages is not affected. Measurement of inositol 1,4,5-triphosphate (IP3) demonstrates that propanil significantly increases the level and the duration of IP3 in macrophages. These results suggest that the inhibitory effect of propanil on macrophage cytokine production is associated with the early stages of LPS mediated signal transduction in macrophages. PMID- 9201268 TI - DNA-PK: at the cross-roads of biochemistry and genetics. PMID- 9201269 TI - A path for coevolution of recombinational DNA repair, transposition, and the common nucleotides. PMID- 9201270 TI - Preferential incision of interstrand crosslinks induced by 8-methoxypsoralen plus UVA in yeast during the cell cycle. AB - Interstrand crosslink (ICL) induction by 8-methoxypsoralen plus UVA and the incision step of the repair have been investigated during the mitotic cell cycle of haploid Saccharomyces cerevisiae. Cells were synchronised by elutriation and events were examined at the level of the MAT alpha and the HML alpha loci in a SIR strain. The DNA sequence of these two loci is identical, but the MAT alpha locus may be replicated earlier in S phase and is transcriptionally active while the HML alpha locus may be replicated later in S phase and is transcriptionally inactive because of Sir repression that creates a heterochromatin-like structure at this locus. ICL were induced to similar extents in both loci during the stages of the cell cycle examined, and these levels were identical to those reported for asynchronous cultures. Preferential incisions occurred for ICL in the MAT alpha locus compared to those in the HML alpha locus, independently of the cell cycle phase studied. The levels of incision were comparable for events in the early G1 phase (eG1), late G1 phase (lG1), early S phase (eS), middle S phase (mS), late S phase (lS) or G2 phase (G2). Thus the preferential incision of ICL observed previously in asynchronous cell culture is maintained throughout the cell cycle and, surprisingly, occurs equally well in G1. Here the opportunities for recombination to further process the incised damaged are substantially limited compared to those in the S and G2 phases. PMID- 9201271 TI - Damage-induced ectopic recombination in the yeast Saccharomyces cerevisiae. AB - Mitotic recombination in the yeast Saccharomyces cerevisiae is induced when cells are irradiated with UV or X-rays, reflecting the efficient repair of damage by recombinational repair mechanisms. We have used multiply marked haploid strains that allow the simultaneous detection of several types of ectopic recombination events. We show that inter-chromosomal ectopic conversion of lys2 heteroalleles and, to a lesser extent, direct repeat recombination (DRR) between non-tandem repeats, are increased by DNA-damaging agents; in contrast, ectopic recombination of the naturally occurring Ty element is not induced. We have tested several hypotheses that could explain the preferential lack of induction of Ty recombination by DNA-damaging agents. We have found that the lack of induction cannot be explained by a cell cycle control or by an effect of the mating-type genes. We also found no role for the flanking long terminal repeats (LTRs) of the Ty in preventing the induction. Ectopic conversion, DRR, and forward mutation of artificial repeats show different kinetics of induction at various positions of the cell cycle, reflecting different mechanisms of recombination. We discuss the mechanistic and evolutionary aspects of these results. PMID- 9201272 TI - Influence of nucleotide excision repair of Escherichia coli on radiation-induced mutagenesis of double-stranded M13 DNA. AB - To investigate a possible role of nucleotide excision repair (NER) of E. coli in the removal of gamma-radiation-induced DNA lesions, double-stranded M13mp10 DNA, which contains a part of the lac operon, including the promoter/operator region, the lacZ alpha gene and a 144 basepair (bp) inframe insert in the lacZ alpha gene, as mutational target was gamma-irradiated in a phosphate buffer under N2. Subsequently, the radiation-exposed DNA was transfected to wild-type or NER deficient (uvrA-) E. coli, mutants in the mutational target selected, followed by characterization of the mutants by sequence analysis. Both the mutations obtained from wild-type and uvrA- E. coli appeared to consist mainly of bp substitutions. However, in contrast to wild-type cells, a relatively large proportion of the mutations obtained from the NER-deficient cells (about 25%) is represented by -1 bp deletions, indicating that NER may be responsible for the removal of lesions which cause this particular type of frameshift. Comparison of the bp substitutions between both E. coli strains showed considerable differences. Thirty per cent of all bp substitutions in the NER-deficient host are T/A-->C/G transitions which are virtually absent in wild-type E. coli. This indicates that NER is involved in the elimination of lesions responsible for these transitions. This may also be true for a part of the lesions which cause C/G-->T/A transitions, which make up 52% of the bp substitutions in uvrA- cells versus 17% in wild-type cells. Strikingly, C/G-->G/C transversions appeared to be only formed in wild-type, where they make up 22% of all bp substitutions, and not in the NER-deficient E. coli. This result suggests, that due to the action of NER, a particular type of mutation may be introduced. A similar indication holds for C/G ->A/T transversions, which are predominant in wild-type (58%) and in the minority in uvrA- cells (15%). PMID- 9201273 TI - The role of N-acetylcysteine as a putative radioprotective agent on X-ray-induced DNA damage as evaluated by alkaline single-cell gel electrophoresis. AB - Samples of human whole blood from 8 different donors were incubated with physiological saline or N-acetyl-L-cysteine (NAC, 1 x 10(-3) M) before being irradiated in vitro with high-energy X-rays (0.7 or 2.0 Gy). Primary DNA damage was evaluated in isolated lymphocytes using alkaline single-cell gel electrophoresis. Whereas the lymphocytes from non-irradiated blood samples showed a similar 'background level' of damage, there was a difference in sensitivity towards the radiation-induced DNA damage, especially at 2.0 Gy. When the data were pooled there was a clear and dose-related increase (p < 0.001) in damage, both in the absence and presence of NAC. Using the two most sensitive 'comet parameters' for DNA damage, i.e., the tail inertia and tail moment, the radiation induced damage was found to be significantly increased already at 0.7 Gy in the samples that had been irradiated without NAC. Overall, NAC was found to be without radioprotective effects. Instead, the incubation with NAC itself was found to be associated with a slightly increased level of DNA damage. If the present findings are relevant also in an in vivo situation using peripheral lymphocytes as a surrogate for non-malignant cells in the body, NAC seems to be of limited value as a radioprotective agent in the clinic, at least when it comes to the acute DNA-damaging effects of therapeutic doses of high-energy X-rays. PMID- 9201274 TI - Incidence and kinetics of distant metastases in patients with operable breast cancer. AB - The purpose of this paper is to evaluate the incidence and kinetics of distant metastases in operable breast cancer and to relate these estimates to various tumor and patient characteristics. The records of 309 consecutive patients with operable breast cancer in stage T1-4N0-1M0 were reviewed, and the incidence of distant metastases (DM) and death due to DM were evaluated. 195 patients had positive axillary nodes with the following distribution of the number of nodes: 45% had 1-2 node, 16% had 3-4 nodes, 14% and 25% had 5-7 and more nodes, respectively. All patients were treated with radical mastectomy with axillary nodes dissection (the only treatment in 39% of cases). In 198 cases radical mastectomy was combined with radiotherapy and/or chemotherapy given pre- or postoperatively. Hormonal treatment was given in 27% of cases. Minimum follow-up was 10 years. Distant metastases were found in 150 cases (49%) and in 78 cases (25%) they developed early, during the first 18 months follow-up. Average rate of DM in N0 cases was 25%. Number of involved nodes and extracapsular invasion were found significant and independent prognostic factors. High risk (50%) of DM and death due to DM correlate with age < 40 y, premenopausal status, tumor stage > or = T3, more than 2 axillary nodes and/or extracapsular invasion. The linearity of the curves for freedom from DM and for freedom from death due to the DM suggest uniform distribution of progression rates with a median value for halving time for freedom from early DM of about 8 months, and of about 40 months for freedom from the DM occurring later than 18 months, being for whole group an average of 20 months. High incidence of DM is a significant cause of poor long-term survival. Early appearance (< 18 month follow-up) of about half of the DM suggests that they are already present as subclinical micrometastases at the time of initial loco-regional treatment. The time of appearance of distant metastases is consistent with a wide range of metastatic cell burdens among patients. Systemic therapy, at least for selected group of patients, might decrease the incidence of DM and improve long-term results. PMID- 9201275 TI - Prognostic parameters in low-grade non-Hodgkin's lymphomas. AB - In a group of 73 patients with low-grade non-Hodgkin's lymphomas (LG NHL) a multivariate analysis of the following variables was performed: pathological types following the International Working Formulation, clinical stage, B symptoms, erythrocyte sedimentation rate, hemoglobin level, white blood cell and lymphocyte count, serum gamma globulin level, serum total lactic dehydrogenase (LDH) level, mitotic and complete remission. The patients with lymphocytic lymphoma manifested the best survival in this group. Low-proliferative lymphomas showed better survival than high-proliferative lymphomas at 3 and 10 years. B symptoms, serum total LDH level and complete remission were significant independent prognostic factors. PMID- 9201276 TI - Clinical thermoradiotherapy: the influence of some prognostic parameters on recurrence-free survival. AB - Thirty-one patients with loco-regional advanced tumors accessible for local thermoradiotherapy were treated at the Institute of Oncology in Ljubljana, between 1989-1993. There were six primary inoperable and 25 recurrent or residual tumors after previous radiotherapy. In 13 patients treatment consisted of combined interstitial water hyperthermia and brachyradiotherapy, in 5 patients combination of interstitial hyperthermia and percutaneous radiotherapy was used, and percutaneous microwave hyperthermia with percutaneous irradiation was employed in remaining 13 patients. Complete response (CR) was achieved in 17/31 (55%) of all treated patients. Among various tumoral and therapeutic parameters tested significant influence on complete response rate was found for tumor volume (p = 0.047), minimum intratumoral temperature (p = 0.004), time interval between hyperthermia and radiotherapy (p = 0.02), and fraction-size of immediate radiotherapy (p = 0.002). More than one hyperthermia treatment and total tumor dose of irradiation > 45 Gy did not significantly improve local control rate in our patients. For all 31 patients treated with thermoradiotherapy 3-year recurrence-free survival (RFS) of 41% was achieved. For the group of 9 patients in whom the interval between hyperthermia and irradiation exceeded 1 hour, RFS of 18% compared to 53% for 22 patients treated with "synchronous" thermoradiotherapy was achieved, however the difference between the groups was not significant (log rank p = 0.17). In 25 patients in whom minimum intratumoral temperature (Tmin50) exceeded 42.5 degrees C significant difference in RFS between the subgroups of 19 patients treated synchronously and 6 patients in whom time interval between the two modalities was longer than 1 hour, i.e. 65% vs. 25% respectively, was found (log rank p = 0.048). However, most favorable RFS of 81% was achieved in the subgroup of 15 patients in whom good hyperthermia treatment (Tmin50 > or = 42.5 degrees C) was followed by an immediate irradiation using fraction size > or = 3 Gy (p = 0.015). Treatment related toxicity was acceptable and did not correlate with response rate. Our conclusion is that thermoradiotherapy is more effective when somewhat larger fraction-size of radiotherapy than conventional, i.e. 3-5 Gy, are employed in synchronous combination of both treatment modalities. PMID- 9201277 TI - In vitro and in vivo studies of murine sarcoma cells after prolonged treatment with promoting phorbol ester TPA. AB - Murine sarcoma cell line (L-1) treated with promoting phorbol ester (TPA) showed decreased content and activity of protein kinase C (PKC) as measured by Western blotting and histone phosphorylation methods. The PKC depleted line (L-1R) produced bigger, tumors after s.c. transplantation into syngeneic mice and more spontaneous and artificial metastases developing after i.v. injection of tumor cells. The in vitro studies revealed decreased: adhesiveness, migratory and invasiveness properties of PKC depleted cells. Negative correlation between in vitro and in vivo studies were found. PMID- 9201278 TI - Growth-regulating influence of nonactivated resident macrophages on transformed and tumor cells in the in vitro contact interactions. AB - Susceptibility to cytostatic activity of nonactivated macrophages (Mph) of Syrian hamster embryo fibroblasts (HEF) transformed in vitro by BAV-3, SV40, RSV-SR, or spontaneously and of their in vivo selected variants was studied in dynamics (5 days of co-incubation). With the use of 3H-TdR incorporation test it was demonstrated that HEF transformed by BAV-3 appeared to be able to overcome the cytostasis at 4-5 day of the co-incubation with Mph, in contrast to deeply suppressed spontaneously transformed cells of STHE strain. HEF transformed by SV40, or RSV-SR appeared to be resistant to growth-inhibiting activity of Mph during almost all the 5-day period. The in vivo selected malignant variants of STHE cells were able to recover from cytostasis after 4-5 days of the co incubation with nonactivated Mph, in contrast to low-malignant variant and to parental cell strain. PMID- 9201279 TI - Retinoic acid-induced alteration of cell surface topography and other changes in an oral carcinoma cell line in culture. AB - Nature of antiproliferative action of retinoic acid (RA) on KB cells was studied by using monolayer and agar culture techniques. RA-treated cultures showed increased requirement of serum for their growth. Growth of colonies in agar culture was significantly retarded when cells were treated with 40 mumol RA. RA induced growth inhibitions in both monolayer and agar cultures were independent of cell seeding densities. Cortisone and hydrocortisone showed no reversal of the inhibitory effects induced by RA on KB cells. Scanning electron microscopy study revealed a significant alteration in cell surface topography of RA-treated cells in monolayer culture. The results demonstrate that RA has a potential of reversing some of the properties which are associated with transformed state of oral carcinoma cells. PMID- 9201280 TI - Glutathione and glutathione metabolizing enzymes in tissues and blood of breast cancer patients. AB - Many reports indicate that glutathione and enzymes cooperating with it are important in neoplastic processes. Glutathione (GSH) concentrations and glutathione S-transferase (GSH STr) and glutathione peroxidase (GSH-Px) activities were determined in breast cancer tissue and adjacent healthy tissues, as well as in blood of 28 patients. There were considerable differences in the investigated parameters among individual patients. Therefore we analyzed the paired samples of normal and cancerous tissues from the same individual. In 68% of the patients the activities of GSH-Px and in 85% patients those of GSH STr were found to be higher in the tumor than in the normal tissue. GSH concentration in 48% tumor samples were higher and in 44% lower than in corresponding normal tissues. Statistically significant correlation was found between GSH-Px and GSH STr in normal (r = 0.51, p < 0.005) and in cancer tissues (r = 0.64, p = 0.001). Correlation coefficient between GSH Px activity in normal and corresponding cancer tissues was r = 0.71 (p < 0.001), however this correlation in the case of GSH STr was much lower but still significant (r = 0.38, p < 0.05). No significant correlation in the determined parameters was found between erythrocytes or plasma and normal or cancer tissues. PMID- 9201281 TI - Phylloid breast tumors and three steroid hormone receptors. AB - The concentrations of three steroid hormones (estrogen, progesteron and 1,25 dihydroxycholecalciferol) receptors (ER, PgR, DR) in tissue cytosol were analyzed in a group of 17 breast phylloid tumors. Comparison with breast carcinoma tissue samples (n = 37) did not reveal significant differences in average values of ER, PgR, and DR. Comparison with another control set of 30 samples of dysplastic tissue of the mammary gland showed significant differences only in PgR values. Only 18% of phylloid tumor samples contained levels above cut-of-line of all three receptors (ER, PgR, DR-5,10,10 resp. fmol/ mg protein). The most frequent combination was ER+PgR+DR-(41%). As far as we know, DR in phylloid breast tumors have never been examined before. In approximately 60% of our samples we found the expression of DR, in 36% the estimated values were above 10 fmol/mg protein. Cells of the tissue not expressing DR seem to belong to a special phenotype. We found no ER+PgR- or ER-PgR-combinations in them. The group which expresses DR is characterized by a higher dispersion of PgR values. PMID- 9201282 TI - Familial testicular cancer and developmental anomalies. AB - Familial occurrence belongs to factors followed in etiology and pathogenesis of testicular germ-cell tumors. Association with abnormal testicular development, or with other risk factors is relatively frequent. In our material 650 patients had been treated for testicular cancer in the period of 1981-1995. Familial occurrence was observed 7-times (1.08%), most frequently in combination with cryptorchidism. Individual families were analyzed in details, including HLA typing. On basis of the observations the supplementation of initial examination of each patient with suspicious testicular cancer with detailed familial history aimed also at the occurrence of urogenital developmental anomalies and tumors has been recommended. The knowledge about familial tumor occurrence in the first degree relatives in combination with thorough testicular self-examination is being considered of great importance in the secondary prevention. PMID- 9201283 TI - Richter syndrome with emphasis on large-cell non-Hodgkin lymphoma in previously unrecognized subclinical chronic lymphocytic leukemia. AB - The authors report seven cases of Richter's syndrome, i.e. of large-cell non Hodgkin's lymphoma (NHL) arising in association with chronic lymphocytic leukemia (CLL). Six patients had the recently recognized variant of this syndrome, occurring in patients with previously undiagnosed subclinical CLL. All patients were treated with aggressive chemotherapy and a complete response of large cell NHL was achieved in 4/7. A complete response of NHL was observed in 3 out of 6, and a partial response in 2/6 patients with simultaneous occurrence of subclinical CLL and large cell NHL (response rate 5/6). Our findings might suggest that patients with Richter syndrome occurring in previously undiagnosed subclinical CLL could represent a better prognostic group in the overall population of patients with large cell NHL transformation of CLL. PMID- 9201284 TI - Possible role of indoor environment and coal combustion emission in lung carcinogenesis in Fuyuan County, China. AB - Fuyuan Country, in Yunnan Province, China has an extremely high lung cancer mortality both in males and non-smoking females. Out of 5768 deaths, 588 patients died of malignant diseases. Lung cancer was the number one cause of death among malignant diseases both in males and females. The rate of lung cancer death to the whole of malignant diseases was 56.2% for males and 55.0% for females. Indoor soot and combustion emission derived from smoky coal produced in northern Fuyuan exhibited high mutagenic activities against Salmonella typhimurium TA98 strain in Ames test. Resected lung tissues derived from the patients with lung cancer in Fuyuan contained significantly higher concentrations of benzo(a)pyrene than those in Japan, both in males and females (i.e., 608.7 +/- 477.1 pg/dry weight for samples of the patients in Fuyuan, 180.1 +/- 104.5 for Japanese non-smokers, and 207.5 +/- 98.8 for Japanese heavy smokers, respectively). These results suggest that mutagenic chemicals contained in coal as well as indoor environment may have a great influence on lung carcinogenesis in Fuyuan, Yunnan Province, China. PMID- 9201285 TI - Increasing occurrence of oropharyngeal cancers among males in Slovakia. AB - The gradual decline of oropharyngeal cancers in postwar period was followed by their rapid increase during recent two decades among males in Slovakia. Overall age-adjusted incidence rates of cancers of oral cavity and pharynx increased from 4.5 in 1968-1970 to 20.7 in 1990-1992 and corresponding mortality rates from 2.8 to 14.0 per 100,000 males. Oropharyngeal cancers have recently accounted for 6.5% of all newly diagnosed cancers yearly and present the fourth most frequent cancer site among males in this country. The cancers of tongue, floor of mouth, oropharynx and hypopharynx are responsible for the dramatic increase of this combined site in males. The culmination of the age-specific incidence and mortality rates of these cancers in the age groups 40-59 confirms the leading role of middle-aged men in their increase and dominant position. The occurrence of cancers of major salivary glands and nasopharynx in males, as well as the incidence and mortality rates of all oropharyngeal cancers in females remained very low and an unchanged (less than 1% of total). Increasing and extremely high incidence and mortality rates from oropharyngeal cancers among males in Slovakia require more effective primary prevention, above all substantial reduction of smoking. PMID- 9201286 TI - Expression of 65-kDa oncofetal protein in experimental hepatoma after anticancer therapy. AB - We have tested the expression of a 65-kDa oncofetal protein (p65) after combined treatment with menadione and methotrexate in hamsters transplanted with Kirkman Robins hepatoma. The treatment of tumor-bearing animals with these compounds significantly inhibited both the tumor development and the expression of p65. This inhibition in tumor tissue was calculated from densitograms of Western blots. The inhibition of p65 expression was also confirmed in the serum of hepatoma bearing animals by using solid-phase radioimmunoassay (RIA) to quantify the specificity of polyclonal antibodies to fetal p65 molecules. Additionally, p65 was shown to localize both in cytoplasm and in the nuclear extracts prepared from hepatoma tissue. PMID- 9201287 TI - Increased efficacy of aphidicolin killing of human neuroblastoma cells in vitro by encapsulation in liposomes. AB - Aphidicolin is a tetracyclic diterpene antibiotic which kills human neuroblastoma cells (NB) in vitro while it has no significant effect on the viability of different human cell types including normal embryonal cells. In the present study, we tested whether aphidicolin encapsulated in liposomes kills NB cells with the efficacy superior to that of unencapsulated aphidicolin. The drug was entrapped in vesicles composed of phosphatidylcholine, phosphatidylserine and cholesterol in a molar ratio of 83:5:12. The treatment with encapsulated aphidicolin at a concentration of 200 nmol for 5 days killed all cells of three human NB cell lines. In contrast, at least 30% of the cells survived 5 days of treatment with 200 nmol unencapsulated aphidicolin. The results showed that aphidicolin killing of human NB cells may be increased by encapsulation in liposomes. PMID- 9201288 TI - Preclinical comparison of bis-diketopiperazine-propane (dexrazoxane) and bis diketopiperazine-ethane (antimet) on the adriamycin-cardiotoxic effect. AB - A cardiotoxic effect induced by adriamycin (by repeated i.v. administration to experimental rats in 7-day intervals of administration) begins to be manifested in the ECG record by prolongation of the S alpha T segment between days 14 and 20, on day 30 it is statistically significant. By means of this index, the known protective effect of dexrazoxane (the preparation Cardioxan) against adriamycin cardiotoxicity has been successfully confirmed in a four-week experiment. A comparative study (using the identical frequency of the dosing scheme and S alpha T segment as the decisive parameter) has revealed that antimet-as another original substance of the diketopiperazines group-also involves (though less significantly) protective effects against the toxic action of adriamycin. PMID- 9201289 TI - Ascorbic acid and 6-deoxy-6-chloro-ascorbic acid: potential anticancer drugs. AB - The role of ascorbic acid (AA) in prevention and suppression of carcinogenesis has been known for a long time. It was also found that AA may inhibit the growth of some tumor cells in vitro and in vivo. We examined the influence of ascorbic acid and 6-chloro-6-deoxy ascorbic acid (6-Cl-AA) on the growth of various human cell lines: lung fibroblasts (Hef), ovarian adenocarcinoma (OVCAR), colon adenocarcinoma (HT-29), laryngeal carcinoma (HEp2) cells, HEp2 cells resistant to vincristine (HEp2VA3), cervical carcinoma (HeLa) cells, HeLa cells resistant to cisplatin (Helacis), breast adenocarcinoma (SK-BR-3) cells, and SK-BR-3 resistant to doxorubicin (SK-BR-3-Dox), as well as mouse fibroblasts L929, mouse melanoma B16 (Mel B16) cells and Chinese hamster fibroblasts (V79). Both drugs arrested the growth of: HeLa, SK-BR-3, SK-BR-3-Dox, L929, and Mel B16 cells, but did not influence the growth of others: Hef, OVCAR, HEp2, HEp2VA3 and V79. 6-Cl-AA suppressed more the proliferation of HeLacis, SK-BR-3-Dox and Mel B16 cells than AA, while AA was active only against HT-29 cells. Inhibitory effect of 6-Cl-AA was confirmed by the in vivo experiments on solid melanoma B16 tumors. Our results indicate that AA and 6-Cl-AA could serve as potential antitumor agents, especially against some tumor cells resistant to chemotherapy. PMID- 9201290 TI - The radiosensitivity of human malignant melanomas evaluated by cytokinesis-block micronucleus assay. AB - Cytokinesis-block micronucleus assay (CB-MNA) was applied for comparison of radiation sensitivity of 25 human malignant melanomas in primary culture. Cells obtained from tumor specimens were irradiated (0-4.Gy) on dishes, incubated with cytochalasin B (2 micrograms/ml) to block cytokinesis, stained in situ and micronuclei (MN) scored in binucleate cells (BNC). Proportions of BNC in nonirradiated controls after fixed time of incubation (96 h) ranged from 2.3 to 38% indicating great differences (C.V. = 74%) in proliferative activity among tumors evaluated. No correlation was observed between proliferative activity and susceptibility of cells to induction of MN by radiation. The great inter-tumor heterogeneity was observed in respect of radiation sensitivity expressed either as normalized (Net) frequency (Fq) of BNC with MN or as number of MN per BNC. Both endpoints differed widely at 2 Gy and 4 Gy as well (Net FqBNC with MN = 0.28 25.4% or 1.5-45% and MN/BNC = 0.004-0.309 or 0.013-0.593 respectively at 2 Gy and 4 Gy) with coefficients of variation ranging from 44 to 57%. Extreme difference in MN frequency was also observed between one primary tumor and its metastasis indicating intra-tumor heterogeneity. Our results suggest that CB-MNA may contribute some clinically useful information for discriminating tumors that will eventually respond to radiotherapy and those that will probably not. However, studies aimed at comparison of MN induction in vitro with clinical radioresponsiveness of malignant melanomas are urgently required. PMID- 9201292 TI - Comparative study of blood insulin levels in breast and endometrial cancer patients. AB - Blood insulin level was measured in 113 breast cancer (BC) patients, 18 endometrial cancer (EC) patients, and 35 women with benign breast disease (BBD), after fasting and after 120 min of oral glucose tolerance test (OGTT). A significant increase in reactive insulin level was shown in postmenopausal BC patients with abdominal obesity (waist/hip ratio > 0.85) and no differences in insulin level were found between BC and BBD patients. Menstrual status and overweight (Quetelet index) did not influence significantly blood insulin concentration in BC patients, but the basal insulin level was lower in those patients who had been moderate smokers. In EC patients, the level of insulin after fasting and following 120 min OGTT was much higher than in BC and BBD patients although they had a similar body mass to these groups of patients. The effect of age on insulin secretion in BC patients is discussed as well as the possible causes and consequences of hyperinsulinemia developing in EC and BC patients. PMID- 9201291 TI - Radiosensitivity of different aged human lymphocytes following electron irradiation in vitro. AB - Cytochalasin B-blocking micronucleus test and chromosomal aberration analysis were used in this study to estimate the yield of individual variability in radiation response of different aged human lymphocytes. Both analyses were performed in three groups of adults, aged 18-65 years, on two sampling times, following irradiation by therapeutical dose of 2 Gy e- in vitro. No statistically significant difference in the induced yield of exchange aberrations between individuals under consideration was found. The yield of total aberration data showed greater variability and was statistically significant in the oldest group against two other adult groups. Regarding to fixation times no statistically significant differences in the induced yield of chromosomal aberrations (exchanges as well as total aberrations) were observed. The study has shown a slight increase in spontaneously occurring micronuclei with age. Almost equal mean number of radiation induced micronuclei was observed in the groups of adults aged 18-20 and 45-55 years. The highest mean number was observed in the oldest group. Evident variation in number of radiation induced micronuclei among individuals from the same age group was observed. The results of micronuclei assay for all individuals under consideration show statistically significant difference in the yield of radiation-induced micronuclei regarding the second fixation time. This study has shown that cytochalasin-B blocking micronucleus test is more sensitive assay than chromosomal aberration analysis for the estimation of individual radiosensitivity. PMID- 9201293 TI - Prognostic value of DNA ploidy in breast cancer stage I-II. AB - The DNA content of paraffin-embedded tumor tissue has been measured by flow cytometry in 169 patients with operable breast cancer Stage I-II. The medium follow-up period was 123 months. Aneuploid primary tumors were found in 49% of patients. Tumor ploidy significantly correlated with histological type of tumor (p < 0.05), whereas no clear correlation between DNA ploidy and tumor size, histological grade and lymph node involvement was found. After 10-year follow-up, recurrence-free survival (RFS) of patients with diploid tumors was slightly better than the survival of those with aneuploid tumors, but the difference was not statistically significant (p = 0.39). In a Cox multivariate analysis only the axillary lymph node involvement and tumor size proved to be independent prognostic factors for recurrence, whereas DNA ploidy lost its prognostic value already in the univariate analysis. Therefore, we can conclude that the information on DNA ploidy, obtained from archival material, does not contribute significantly to better discrimination between good-risk and poor-risk operable breast cancer patients. PMID- 9201294 TI - Differential chemosensitivity of leukemic cells in the myeloid and lymphoid phases of stem cell leukemia (a case report). AB - A five-year-old girl, initially diagnosed as having acute lymphoblastic leukemia (ALL; FAB-L1) relapsed with ALL 4 months after completion of chemotherapy (BFM 83). The initial ALL presentation and subsequent ALL relapse were analyzed using conventional morphology, cytochemistry, cytogenetics and immunophenotyping. The results were consistent with a diagnosis of B-lymphocyte precursor ALL. Bone marrow leukemic cells revealed a 46, XX karyotype at diagnosis and a 46, XX, del(7) (q22; qter) when the girl first relapsed. The case was managed with a BFM REZ-ALL 90 protocol. Upon completion of the first cycle of the protocol, severe myelosuppression developed. This was treated with GM-CSF. Three days later, however, GM-CSF was stopped because the WBC reached 1.1 x 10(9) per liter with 60% of blasts in peripheral blood. Laboratory characteristics were typical of AML. Cytogenetic analysis revealed 46, XX, del(7) (q22; qter) karyotype as before. The bcr-abl fusion gene was not detected. Myeloid blasts were placed in a culture and maintained at 37 degrees C and 7.5% CO2 for two weeks. During this period, formation of hemopoietic colonies was observed and subsequently analyzed using histology and electron microscopy. This showed that the colonies consisted of differentiating erythroid, megakaryocytic and myeloid cells. Further, the chemosensitivity of leukemic cells was examined in both "lymphoid" and "myeloid" relapse instances. While the "lymphoid" phenotype was characterized by good sensitivity to corticosteroids, a typical feature of the "myeloid" phenotype was a high resistance to corticosteroids with marginally increased sensitivity to ARA C. PMID- 9201295 TI - Adrenal incidentalomas--analysis of 23 cases discovered by ultrasound. AB - Frequent use of abdominal ultrasonography (USG) increases discovery of incidental adrenal tumors. Our experience and concise review of recent opinions on management of adrenal incidentalomas is presented. In four out of 23 patients with adrenal incidentalomas false positivity of USG was found (all on the left side), 4 cases were identified as pseudoadrenal masses. Hormonal activity was proved in 4 out of 15 true adrenal masses (2 pheochromocytomass, 2 aldosteronomas). Five out of 11 hormonally inactive tumors were benign adenomass, 2 myelolipomas, 2 simple cysts, 1 metastasis of bronchogenic carcinoma and 1 tuberculotic involvement. The smallest tumor was aldosteronoma (2 cm in diameter), the largest was myelolipoma (more than 10 cm). Size of benign adenomas ranged between 2.5-4.8 cm. Three main ultrasonic patterns of adrenal tumors were recognized: (1) anechogenic cysts, (2) complex but predominantly hyperechogenic myelolipomas, (3) hypoechogenic all other masses. PMID- 9201296 TI - Psoriasis vulgaris, streptococci and the immune system: a riddle to be solved soon? PMID- 9201297 TI - The SH3 domain of Bruton's tyrosine kinase interacts with Vav, Sam68 and EWS. AB - Bruton tyrosine kinase (BTK) is a cytoplasmic protein tyrosine kinase which controls crucial steps of differentiation of B lymphocytes. Mutations affecting either the PH, SH3, SH2 or kinase domain of BTK all give rise to X linked agammaglobulinaemia (XLA) in humans. In this study, the authors report that the BTK-SH3 domain binds to a set of proteins expressed in pro-B, pre-B and B cell lines. Three of them were characterized as Vav, Sam68 and EWS. The authors show that a Pro-->Leu substitution in a region of the SH3 domain, which is deleted in an XLA patient, is sufficient to abolish BTK-SH3 binding potential. The authors also report that several of the BTK-SH3 binding proteins, including Sam68, EWS and Vav, are tyrosine phosphorylated in conditions that also promote BTK kinase activity. For EWS and Sam68 this tyrosine phosphorylation was cell cycle dependent. PMID- 9201298 TI - A human in vitro granuloma model using heat killed Candida albicans cells immobilized on plastic culture wells. AB - A new model for studying the initial events of granuloma formation in vitro is presented using heat killed Candida albicans immobilized on the surface of plastic culture wells. Human monocytes were induced to accumulate and to proliferate, forming multinucleated giant cells (MGC) and epitheloid cells within 4 days of culture. Tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta and IL-6 were detected in culture supernatants. These monokines, and additionally macrophage colony stimulating factor (M-CSF), were also detected immunocytochemically. The granuloma formation was inhibited by Dexamethasone (Dex), Pentoxifylline (POF), or interferon-gamma (IFN-gamma) in a dose-dependent manner. Antibodies to M-CSF reduced the granuloma formation to a great extent with a striking reduction of monocyte proliferation. Using antibodies to TNF alpha the authors found a complete inhibition of the granuloma including MGC formation and monocyte proliferation. PMID- 9201299 TI - Restoration of MHC class I surface expression and endogenous antigen presentation by a molecular chaperone. AB - Presentation of cytosolic peptides in the context of major histocompatibility complex (MHC) class I antigen is crucial for immune recognition of virus-infected and malignant cells. This process, which is often defective in cancer cells, involves a series of cellular events which may be facilitated by heat shock proteins (molecular chaperones). To address the influence of chaperone function on the presentation of cytosolic peptides, we have utilized B16 melanoma cells (H 2b). These tumour cells are resistant to lysis by MHC class I-restricted cytotoxic T lymphocytes (CTL), due to a very low level of surface MHC expression. The authors found that stably transfected clones of B16 expressing a heterologous heat shock protein (Hsp65) exhibit significantly increased levels of MHC class I antigens on their surface, and are effectively lysed by alloreactive CTL. These MHC class I molecules can form functional MHC-peptide complexes which are recognized by virus-specific CTL. Moreover, mice immunized with Hsp65-expressing tumour cells, but not with control-transfected tumour cells, display a significantly increased resistance to a subsequent challenge with live, wild-type B16. Together, these results indicate that the suitable expression of a molecular chaperone can overcome a defect in MHC class I expression and antigen presentation, and suggest a novel approach to cancer immunotherapy. PMID- 9201300 TI - Cytokine genes are down-regulated when attachment of Entamoeba histolytica to HT 29 colon epithelial cells is prevented. AB - Entamoeba histolytica can cause invasive disease by disruption of the intestinal barriers and subsequent lysis of the intestinal cells. Adherence to and contact dependent killing of host cells requires the galactose inhibitable lectin. To elucidate the mechanism whereby E. histolytica influences host defence, the authors assessed the change of proinflammatory cytokine genes expressed by colon epithelial cells in response to co-culture with E. histolytica trophozoites and carbohydrates, including galactose, N-acetyl-galactosamine or N-acetyl lactosamine, which prevented E. histolytica from attaching to epithelial cells. After HT-29 human colon epithelial cells were co-cultured with E. histolytica trophozoites in the presence or absence of carbohydrates (0.1-100 mM), RNA was extracted from the epithelial cells by an acid guanidinium thiocyanate-phenol chloroform method. Cytokine gene expression was assessed by quantitative RT-PCR using a synthetic internal standard, and proteins were determined by ELISA. IL-8 mRNA expressed by HT-29 cells in response to E. histolytica trophozoites was downregulated in the presence of galactose, N-acetylgalactosamine or N-acetyl lactosamine (0.1-100 mM), and this was paralleled by decreased IL-8 protein secretion. GM-CSF and IL-1 alpha/beta mRNAs were also downregulated in those cells in the presence of these agents. These results suggest that the expression of proinflammatory cytokine genes could be inhibited by preventing E. histolytica from attaching to the host's colon epithelial cells. PMID- 9201301 TI - A cryopreservation method of human peripheral blood mononuclear cells for efficient production of dendritic cells. AB - The establishment of a cryopreservation method for unstimulated fresh peripheral blood mononuclear cells (PBMC) with nearly 100% viability would greatly contribute to the conduct of various immunological experiments. The cells most sensitive to freezing and thawing procedure seem to be dendritic cells (DC) and their precursors, which are of the most potent antigen-presenting cells. The authors investigated and established a method of cryopreserving fresh PBMC from which DC were recovered and differentiated efficiently by using recombinant (r) GM-CSF and rIL-4. PBMC frozen in the presence of 12% dimethylsulfoxide and 25-30% fetal calf serum recovered DC as efficiently as freshly obtained PBMC. Established DC could also be cryopreserved in the presence of 12% DMSO with their viability maintained at more than 90%. The 12% DMSO freezing solutions were superior to both the 10% DMSO solution and the previously reported DC freezing medium (2 M or 15.4% DMSO). The DC obtained from the cryopreserved PBMC expressed HLA-DR, HLA-DQ, CD80 and CD86 antigens, and stimulated allogenic PBMC to an extent almost identical to that obtained from fresh PBMC. These findings indicate that the conditioned medium utilized here enables safe cryopreservation of DC and DC precursors in PBMC. PMID- 9201302 TI - High dose IL-2-activated murine natural killer (A-NK) cells accumulate glycogen and granules, lose cytotoxicity, and alter target cell interaction in vitro. AB - Activated natural killer (A-NK) cells, defined by immunophenotype and selected by adherence to the plastic, were cultured from murine splenocytes for up to 10 days with the addition of 1000 U/ml of recombinant human IL-2 at 48 h intervals. During culture days 2-4 with high DNA synthesis the initially non-granulated small cells established large granular lymphocyte (LGL) morphology and then differentiated further into giant hypergranulated cells with huge accumulations of glycogen. Timed EM observations indicated that specific dual-compartment (lytic) granules arose by a sequence of events starting with neo-synthesis of small progenitors with a dense core and a few membranous lamellae at one pole. Core and vesicular regions probably expanded independently to give the mature organization of the granule. Eventually, the vesicular region of granules contained large amounts of multi-lamellar material and probable debris, and the dense core could be multiplied. Intracellular proteoglycans, visualized with Cupromeronic Blue cytochemistry, were organized in a three-dimensional network within the dense cores. In contrast with earlier reports, and in spite of several fold increased granularity, the in vitro cytotoxicity of the A-NK cells against YAC-1 and B16 cells decreased after the third day of culture. A-NK cells with glycogen accumulations caused focal clearing in melanoma monolayers whereas younger effectors adhered to the targets. It is concluded that high dose IL-2 stimulation causes more far-going progressive morphological and functional differentiations of the A-NK cells than has previously been observed with bearing for the use of these cells in experimental adoptive immunotherapy. PMID- 9201303 TI - Evaluation of T cell subsets by an immunocytochemical method compared to flow cytometry in four countries. AB - The authors tested an alternative method for CD4 and CD8 T lymphocytes enumeration, the immunoalkaline phosphatase method (IA), in three African countries and in Denmark. The IA determinations from 136 HIV antibody positive and 105 HIV antibody negative individuals were compared to the corresponding results obtained by flow cytometry (FC) performed in the respective countries. The authors found good correspondence between the two methods for measurements of CD4 and CD8 T lymphocytes independent of serological status and geographical site. However, the CD4 and CD8 T lymphocytes values obtained by the two methods are not interchangeable as IA compared to FC consistently gives higher percentage of CD4 T lymphocytes, and lower percentage of CD8 T lymphocytes. Mean differences between the two methods did not differ between the three African countries indicating that the IA method provides systematic results. Replicate measurements suggested good correspondence between results obtained by IA. By using an IA level of < 300 CD4 T lymphocytes/microliter, the sensitivity was 81% and specificity 96% for detecting an FC level of < 200 CD4 T lymphocytes/microliter. Using an IA level of < 20% CD4 T lymphocytes, the sensitivity was 89% and specificity 95% for detecting an FC level of < 14% CD4 T lymphocytes. The FC and IA methods had the same internal correspondence between low absolute CD4 T cell count and low CD4 percentages; the sensitivity and specificity for detecting a low absolute CD4 T cell counts with a low CD4 percentage was 92% and 68% for FC and 91% and 73% for IA, respectively. The IA method is 10-fold cheaper than FC, is independent of advanced laboratory facilities, and does not need immediate processing of samples as blood smears can be stored for long periods. The IA method is therefore suitable for use in areas with limited resources and laboratory facilities where there is a need for immunological surveillance in hospital or community studies. PMID- 9201305 TI - B cell memory in xid mice is long-lived despite reduced memory B cell frequency. AB - Brutons tyrosine kinase (Btk) deficient xid mice have a diminished primary T cell dependent immune response, resulting in a reduced memory B cell frequency. Boosting at 35 days post primary immunization, however, generates a normal secondary immune response, indicating a functional memory B cell compartment. The longevity of B cell memory appears to depend on both the presence of antigen and expression of cell survival genes such as bcl-2. Since there is a natural decay in the number of memory B cells over time and since xid B cells have been demonstrated to have reduced Bcl-2 levels, we aimed at determining whether B cell memory of xid mice would be long-lasting. This report demonstrates that memory B cell precursors are detectable in xid mice more than 100 days after primary immunization. Furthermore, a secondary immune response of normal magnitude and kinetics can be generated in xid mice at 150 days after primary immunization indicating that B cell memory is long-lived in xid mice. Thus, although survival of B cell memory is presumably dependent on immunoglobulin (Ig)-mediated interaction with antigen, this interaction does not depend solely on signalling through Btk. PMID- 9201304 TI - Retrovirally induced mouse anti-TCR monoclonals can synergize the in vitro proliferative T cell response to bacterial superantigens. AB - Antibodies directed against the beta chain of the T-cell receptor (TCR) have been detected in animals and in humans in a number of distinct immune states that do not involve direct immunization with either T cells or TCR epitopes. When C57B1/6 mice are infected experimentally with the LP-BM5 retrovirus mixture they produce increased titres of autoantibodies directed against TCR V beta complementarity determining region 1 (CDR1) epitopes. Here, the authors utilized hybridoma technology to isolate monoclonal immunoglobulin (Ig)M antibodies (MoAbs) that arose at the peak of infection. The authors characterized the binding specificity tested using synthetic peptides modelling the CDR1 segments of 24 distinct V beta gene products and determined the VH gene usage by two such monoclonals. One binds to a restricted set of TCR V beta CDR1 peptides, and the second reacts with approximately half of the CDR1 peptide homologues. These MoAbs are specific for T cell receptor beta chains and do not bind to immunoglobulin light chains or to unrelated protein molecules. Both MoAbs bind to intact T cells expressing the V beta domain (human V beta 8 and mouse V beta 11) from which selection peptides were derived, and costimulate a V beta specific in vitro T cell proliferative response induced by the staphylcoccal enterotoxin E (SEE) superantigen. PMID- 9201306 TI - Differential role for IL-2 and IL-15 in the inhibition of apoptosis in short-term activated human lymphocytes. AB - Interleukin (IL)-15 is a newly described cytokine with properties similar to IL 2. Even though it does not share sequence homology with IL-2, both cytokines bind to the same receptor with the noted exception of a cytokine specific alpha-chain. In this study the authors compared IL-2 and IL-15 to determine their ability to rescue short term activated lymphocytes (phytohaemagglutinin stimulation of peripheral blood mononuclear cells for 6 days, followed by expansion in medium containing IL-2 for 2 days) from apoptotic cell death. The authors found that both IL-2 and IL-15 can inhibit induction of apoptosis in this experimental model with similar time and dose kinetics. On mRNA or protein levels induction of pro- and anti-apoptotic gene products like fasL, bcl-2, or bax with minor effects on fas/Apo-1 or bcl-XL was observed under culture conditions with both IL-2 and IL 15. Next, it was found that phytohaemagglutinin (PHA) blasts were less responsive (in terms of cellular proliferation and prevention from apoptosis) to IL-2 if signals through the alpha-chain were blocked, with no effect on beta-chain specific monoclonal antibodies (MoAb). By contrast, IL-15 was less effective in induction of cellular proliferation and prevention of apoptosis if IL-2R beta chain specific MoAb were added to cell cultures. Testing intracellular signalling induced by IL-2 or IL-15, the authors found identical changes in tyrosine phosphorylation patterns in PHA blasts cultured in medium or under IL-2 or IL-15 stimulation. By contrast, they found consistent differences if PHA stimulated peripheral blood mononuclear cells (PBMC) were expanded in medium containing IL 15 (instead of IL-2). These IL-15 expanded PHA blasts showed a significantly increased percentage of apoptosis after growth factor withdrawal. Furthermore, IL 2 was more efficient than IL-15 in rescuing IL-15 expanded PHA blasts from apoptosis. In IL-15 expanded PHA blasts expression of IL-2R alpha-chain was lower than that in IL-2 expanded PHA blasts. A model presenting a differential role for IL-2 and IL-15 in inhibition of apoptosis in vivo is discussed. PMID- 9201307 TI - Arthritis susceptibility in mice expressing human type II collagen in cartilage. AB - Collagen type II (CII) induced arthritis (CIA) in mice is an experimental model for rheumatoid arthritis. Induction with non-self (e.g. human) CII induces severe arthritis whereas the mice are less susceptible to induction with self CII (i.e. mouse). To analyse whether an autoimmune response to human CII can develop and is pathogenic the authors have established transgenic mice expressing human CII in cartilage and backcrossed them into two different gene backgrounds susceptible to CIA (DBA/1 and C3H.Q). The transgenic human CII expression was restricted to cartilage and did not disturb cartilage morphology or lead to chondrodystrophy. In addition, development of stress-induced arthritis was not affected by the transgene. The cartilage specific expression of human CII reduced, but did not eliminate, the susceptibility to CIA irrespective of the species source (human, bovine, chick, rat) of CII used for immunization. A common denominator between these heterologous CII in comparison with mouse CII is the previously defined CII 256-270 epitope. An expression level dependent T-cell tolerance was seen in this epitope as well as to the entire CII. However, all human transgenic mouse lines could still mount significant autoreactive T- and B-cell responses. Approximately 10% of the transgenic mice developed arthritis after immunization with human CII. These findings show, therefore, that cartilage-located human CII induce tolerance but can nevertheless be a target for development of arthritis. PMID- 9201308 TI - Conversion of in vitro cultured human monocytes into effective presenters of an HER2/neu-encoded CTL peptide epitope. AB - Tumour-derived peptides have been surveyed, in a variety of systems, for their ability to elicit cytokine release from class I restricted T cells. Analogous studies on ovarian carcinoma have employed the antigen-processing defective T2 cell line, Purified dendritic cells (DC) have been reported to act as highly effective APC. A facile method was developed whereby DC-like cells were generated from monocyte precursors. Herein, evidence is presented suggesting DC-like cells are superior to T2 with respect to their ability to present a defined CTL epitope associated with ovarian carcinoma. PMID- 9201309 TI - Biological effects of the immunomodulator beta 1-3D polyglucose are strongly potentiated by conjugation to biodegradable microbeads. AB - It is demonstrated that the biological effects of the immunomodulator beta 1-3D polyglucose, when covalently linked to polymethacrylate or biodegradable albumin microbeads, are strongly potentiated. The potentiation is recorded as an increased protection effect of the conjugates in Escherichia coli sepsis in mice, and as increased IL-1 production by murine macrophages in vitro. PMID- 9201310 TI - Circulating T cells of patients with active psoriasis respond to streptococcal M peptides sharing sequences with human epidermal keratins. AB - Psoriasis is a T-cell mediated inflammatory skin disease which has been associated with group A, beta-haemolytic streptococcal infections. Four 20 a.a. long M6-peptides sharing 5-6 a.a. sequences with human epidermal keratins were identified. To investigate the role of potentially cross-reactive T cells in the pathogenesis of psoriasis, interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) responses of circulating T cells to these peptides were analysed by ELISPOT and RT-PCR in 14 psoriatic patients, 12 healthy individuals and six patients with atopic dermatitis (AD). Untreated psoriatic patients' responses were significantly higher to these peptides than healthy and AD controls, while responses to a control M6-peptide, not sharing sequences with keratin, were negligible in all groups. No difference was found in response to streptokinase/streptodornase (SK/SD). M6-protein and peptides exclusively elicited IFN-gamma production, with little IL-4 production, even in AD patients. Interferon-gamma responses to all the M6-peptides were abolished after successful treatment of psoriatic patients, but responses to SK/SD were unaffected. The results indicate that active psoriasis is associated with Th1-like cells responding to streptococcal M6-peptides sharing sequences with human epidermal keratin. This is consistent with the hypothesis that psoriasis may be induced and exacerbated in susceptible individuals by M-protein specific Th1-like cells that cross-react with human epidermal keratin. PMID- 9201311 TI - Differential immunological aberrations in patients with primary and secondary Sjogren's syndrome. AB - The aim of the present study was to analyse possible differences in immunological features between patients with primary and secondary Sjogren's syndrome (SS). Ten patients with primary SS and 10 patients with secondary SS also suffering from rheumatoid arthritis, were identified according to established criteria for SS. Ten healthy, age-matched women served as controls. The authors analysed the phenotypic characteristics of lymphocytes in peripheral blood as well as in focal inflammatory infiltrates of minor salivary gland biopsies. Functional analyses of T lymphocytes were performed after stimulation with mitogens and antigen. B cell activity was determined at the single cell level by spontaneous and mitogen induced immunoglobulin production. Serum levels of IL-4, IL-6 and IFN-gamma were also analysed. Patients with primary SS displayed a significantly higher degree of salivary gland inflammation and reduced salivary flow than did patients with secondary SS. Decreased in vitro T cell responses to antigen and mitogens were evident in both patient groups. The CD4/CD8 ratios in both peripheral blood and salivary gland lesions were significantly lower in primary SS compared with secondary SS patients. Polyclonal B cell activation, measured as the frequency of spontaneous immunoglobulin producing cells, was most prominent in primary SS, whereas a diminished response to poke-weed mitogen (PWM), a T cell dependent B cell mitogen, was more pronounced in secondary SS. The results reveal certain immunological aberrations in the whole group of patients with SS. In addition, the authors demonstrated distinct differences in immune dysfunction between patients with primary and secondary SS, indicating that they may constitute separate entities. PMID- 9201312 TI - CD40 ligation inhibits IL-2 and SAC+IL-2 induced proliferation in chronic lymphocytic leukaemia cells. AB - This paper reports on differences between B cell-type chronic lymphocytic leukaemia (B-CLL) and normal B cells in their response to IL-2+thioredoxin (Trx), Staphylococcus aureus Cowan strain 1 (SAC)+IL-2+Trx, and to CD40 ligation of IL 2+Trx and SAC+IL-2+Trx stimulation. The authors found that Trx acted synergistically with IL-2 and SAC+IL-2 in inducing DNA synthesis in B-CLL cells, but not in the normal B cells. Interestingly, IL-2+Trx alone was found to induce proliferation in B-CLL cells from patients with advanced stages of disease. In addition, we also found that IL-2+Trx and SAC+IL-2+Trx-induced DNA synthesis of B CLL cells was inhibited by CD40 activation (by soluble anti-CD40 MoAb and anti CD40 MoAb presented on irradiated CD32L cells). In clear contrast, SAC+IL-2+ Trx induced DNA synthesis of normal B cells was not inhibited by soluble anti-CD40 MoAb. The authors therefore conclude that B-CLL cells differ from normal B cells in their response to IL-2 (IL-2+Trx) and CD40 ligation. PMID- 9201313 TI - Expression and signal transduction of T-cell antigen receptor (TCR)/CD3 complexes on fresh or in vitro expanded T lymphocytes from patients with Hodgkin's and non Hodgkin's lymphomas. AB - T-cell responses against soluble antigens, alloantigens and mitogens are frequently diminished in patients with certain types of cancer. In the present study, the authors investigated possible mechanisms for the partial T-cell immunodeficiency in patients with Hodgkin's or non-Hodgkin's lymphomas. It was found that T-cells from lymphoma patients had significantly reduced proliferative responses to EBV-transformed B-cell lines and to anti-TCR/CD3 MoAb; a 30-50% reduction of cells expressing membrane T-cell receptor (TCR) complexes; and a significantly reduced signal transduction function. Long-term in vitro culture conditions were developed to expand T cells in TCR/CD3-dependent or TCR/CD3 independent manners. With such methods, it was found that the decreased T-cell responses in patients with Hodgkin's and non-Hodgkin's lymphomas appeared to be an intrinsic T-cell defect (not at the antigen presenting cell level), and the T cell responses could be recovered after only a few days in culture. Thus, it is suggested that the T-cell response-defect in Hodgkin or non-Hodgkin lymphoma patients is a reversible phenomenon, dependent on the patient's tumour-bearing environment. PMID- 9201315 TI - Effects of chronic treatments with adrenal steroids on acid-base homeostasis in the rat. AB - Urinary parameters related to acid base homeostasis were studied in adrenalectomized rats (ADX) as well as in ADX treated with physiological doses of corticosterone (B), aldosterone (aldo) or 18-Hydroxycorticosterone (18HOB) during 1, 3 or 5 days, under basal conditions and after gravage with 200 mM HCI. The results showed: a) persistent effect of B and 18HOB increasing titratable acidity principally in response to acidosis; b) an increased phosphate elimination in acidotic B treated ADX on the first day, and in 18 HOB treated ADX on days 3 and 5; c) pronounced increases in blood pH and blood bicarbonate levels provoked by the three steroids on day 1; d) increments of ammonium elimination in response to acidosis by aldo treatments on the first day, while B and 18HOB increase ammonium elimination under almost all conditions during the whole experiment; e) the effects of B and 18 HOB would be independent of an increase in sodium retention as well as glomerular filtration rate. PMID- 9201314 TI - Dual-receptor T cells expressing one self-restricted TCR. AB - During thymic development, T cells rearrange and express genes encoding clonotypic T-cell receptors (TCR), and undergo selective events involving interactions between TCR and thymic major histocompatibility complex (MHC) molecules. Recent studies indicate that up to 30% of peripheral T cells may express two TCR, that both TCR on these cells are functional, and that dual specific T cells can promote autoimmunity. Here we demonstrate that dual-receptor T cells are readily generated in class II-deficient (class II-) mice expressing the class II-restricted AND TCR transgene (TCRtg). While this TCRtg is unable to promote positive selection in class II- mice, T cells arise in class II-TCR+tg mice by co-expressing non-transgenic endogenous TCR that permit positive selection on class I molecules. Our findings indicate that when a rearranged TCR fails to promote positive selection on self MHC molecules, expression of a second receptor can rescue the developing T cell. Accordingly, many dual-receptor T cells may actually be mono-specific in vivo, possessing only one self MHC restricted TCR, and therefore should not require unique regulatory controls to prevent autoreactivity. PMID- 9201316 TI - Water permeability properties of the human small intestine in vitro: effects of Escherichia coli heat-stable enterotoxin. AB - The net absorptive water flux (Jw), the transepithelial potential difference (PD) and the short-circuit current (Isc) were simultaneously measured in the human small intestine in vitro with the following results: 1) An absorptive Jw was observed when the jejunum or the ileum were mounted between two identical standard solutions in the presence of an hydrostatic pressure gradient (delta P) of 13 cm of water (mucosal side positive). 2) The absorptive Jw was a linear function of the applied delta P or the imposed osmotic transepithelial gradient (delta Osm) in both intestinal segments. The hydrostatic (Phydr) and osmotic (Posm) permeabilities to water for jejunum and ileum were: 0.349 +/- 0.049 cm/s vs. 0.156 +/- 0.022 cm/s and 0.0012 +/- 0.0001 cm/s vs. 0.0019 +/- 0.0003, respectively. 3) A fraction of this absorptive Jw was independent of the presence of any hydrostatic, osmotic or chemical gradient and represented the transport associated to movement of water (Jwt). 4) PD and Isc values were similar in the jejunum and in the ileum but the transepithelial resistance (Rt) was significantly greater in ileum than in jejunum. 5) 2 micrograms/ml of E. coli heat-stable enterotoxin (STa) caused a significant inhibition of the absorptive Jw without modification of Phydr, Posm or Isc. 6) After STa treatment, the absorptive Jwt reverted to a secretory one in the jejunum. In the ileum, STa action caused a 48% decrease in the absorptive Jwt values. PMID- 9201317 TI - Influence of the pituitary gland on the immune response in young rats. AB - The response of hypophysectomized (HYPOX) and sham-operated (S-HYPOX) female and male Wistar young rats (8 weeks old) to antigenic stimulation was compared. Humoral antigenic responses against hemocyanin were measured by ELISA. [3H]thymidine incorporation into cultured spleen cells was used to determine proliferative response to concanavalin A (ConA) or antigenic stimulation. Anti hemocyanin serum titers in the HYPOX animals was about half of that observed in control S-HYPOX rats. Similarly, the cellular proliferative response was significantly decreased in HYPOX animals when compared to S-HYPOX rats; the blastogenic response to hemocyanin in UC rats (which did not receive the antigen injection) was close to zero. S-HYPOX control rats responded to direct ConA stimulation as UC controls. Body weight and the weight of pituitary target organs (adrenal, thyroid, ovary and testes) was about 1/4 of that of controls. Hypophysectomy also resulted in a striking reduction in spleen weight. These results indicate that the pituitary gland is involved in cellular and humoral immune regulation in young rats. PMID- 9201318 TI - Visually-evoked pattern and photomyoclonic responses in video game and television epilepsy: case reports. AB - This research paper reports a case study of two male photosensitive epileptic patients, aged 14 and 16 years old respectively, whose epileptic seizures were often triggered by the flickers from television and video games respectively. The 14-year old patient had no family history of epilepsy, while the 16 year old had a family history of epilepsy. A comprehensive electroencephalogram (EEG), including hyperventilation, intermittent photic stimulation (IPS) and pattern stimulation were carried out on them and EEG abnormalities including photoparoxysmal responses (PPR) and generalized myoclonic responses were evoked. A thorough analysis of the EEG morphology of the myclonic responses and the clinical manifestations showed evidence of two separate entitles of seizures namely: visually evoked pattern-myoclonic responses (PTMR) and visually evoked photomyoclonic responses (PMR). PTMR was independent of flash rate and occurred before a PPR and at the same time as the flash rate, while PMR occurred after the PPR and was dependent on flash rate. These findings suggest that "Video Game" epilepsy is probably a pattern sensitive epilepsy, electronic screen being the source of the triggering patterns; hence, the morphology and the family histories and the myoclonic phenomena differ from those of pure photosensitive epilepsy. PMID- 9201319 TI - Alterations in dense LDL subfractions in normolipidemic IDDM patients. AB - Low density lipoproteins (LDL) of human plasma, consist of a continuum of particles subclasses with distinct physicochemical, immunological and hydrodynamic characteristics. Such structural differences are intimately linked to atherogenesis. The current study was designated to investigated the LDL subclasses profile in 12 normolipidemic IDDM patients, and compare it with 11 healthy controls. Four plasma LDL subfractions were isolated by sequential ultracentrifugation in the density range (1.025-1.063 g/ml) and were characterized by their content of free and esterified cholesterol, triglycerides, phospholipids, and proteins. The net electrical charge was evaluated. Plasma concentration of the two denser LDL subfractions were higher in IDDM patients vs control subjects, due to an increase in cholesterol (free and esterified) and phospholipids, while more buoyant subfractions in the two groups were not different. In IDDM patients the LDL profile was skewed towards the dense subclasses LDL-III and LDL-IV, being significant this increase for LDL-IV: 22.3 +/- 5.2 vs 18.3 +/- 4.0%, p < 0.05, X +/- DS. In the healthy controls the LDL profile was skewed toward the lighter subclasses (LDL-I and LDL-II), being significant for LDL-II: 30.0 +/- 4.3 vs 23.3 +/- 4.2%, p < 0.005. Diabetic patients, even those who are normolipidemic, present increased risk of premature atherosclerosis. This suggest that normal values in lipid and lipoprotein profile can mask deeper alterations, such as changes in the composition and distribution of the denser subclasses, whose characteristics make them potentially more atherogenic. Despite the apparently normolipidemic status, dense LDL particles considered to be atherogenic, are increased in IDDM patients. PMID- 9201320 TI - Effects of alpha-MSH and cholinergic agents on cGMP and IP3 levels in rat brain slices. AB - On one hand, it has been demonstrated that the exposure of rat brain slices containing caudate putamen and accumbens nuclei to alpha-MSH brings about an increase in cAMP. This increase is affected when dopamine is present in the incubation medium. On the other hand, an interaction of melanotropinergic-like peptides with acetylcholinergic drugs has been showed to be similar to the one observed with dopamine. In this study we have intended to measure cGMP or IP3 in response to alpha-MSH, and also to study the interaction with cholinergic drugs by measuring the second messengers recently mentioned. cGMP and IP3 have been measured in tissues and medium in their response to the effect of alpha-MSH alone or in the presence of the peptide plus pilocarpine (selective muscarinic agonist) or atropine (selective muscarinic antagonist). None of them modified the cGMP levels when compared with the control group. The exposure of rat brain slices containing CP and Acc nuclei to alpha-MSH resulted in an increase in IP3 levels. Pilocarpine by itself brought about an increase of IP3 only when the highest doses was used. Atropine did not modify the IP3 content. However, when slices were exposured to both alpha-MSH and pilocarpine, IP3 content was similar to control values. The blockage of the muscarinic receptor with atropine blocked the IP3 increase induced by alpha-MSH as well. Therefore, we assume that alpha-MSH does not induce changes in cGMP but it does change the IP3 levels, probably acting at the muscarinic receptor level. PMID- 9201321 TI - Converting enzyme inhibition and renal function of conscious uninephrectomized rats [correction of enzyme]. AB - We studied the effect of a Converting Enzyme Inhibitor, Captopril, on renal function of conscious, chronically instrumented uninephrectomized rats three weeks after surgery and normal two kidney rats. Captopril increased glomerular filtration rate from 9.28 +/- 0.50 to 14.23 +/- 1.07 ml/min.kg and effective renal plasma flow from 31.6 +/- 2.4 to 45.8 +/- 3.7 ml/min.kg in two kidney rats. Glomerular filtration rate and effective renal plasma flow of uninephrectomized rats did not change after acute converting enzyme inhibition (from 6.96 +/- 0.61 to 7.16 +/- 0.39 and 24.5 +/- 2.2 to 27.9 +/- 1.5 ml/min.kg respectively). Plasma renin activity was lower in uninephrectomized rats (0.72 +/- 0.15 ng AngI/ml.h) than in intact rats (1.41 +/- 0.20 ng AngI/ml.h). Acute converting enzyme inhibition increased urinary sodium excretion, fractional sodium excretion and plasma renin activity in both, two kidney and uninephrectomized rats without changes of mean arterial pressure. Present data suggest that the Angiotensin II does not participate in the control of glomerular filtration rate and effective renal plasma flow of uninephrectomized rats but it is implicated in the control of Na homeostasis. PMID- 9201322 TI - [Miguel Dias Pimenta (1661-1715) and the history of chagasic megaesophagus and megacolon]. AB - In the present study an attempt was made to analyse from a clinical viewpoint the descriptions in the book "Noticias do que he o achaque do bicho" by Miguel Dias Pimenta (1661-1715), which are considered by some authors to be the first reference to the chagasic megaesophagus and megacolon that appeared in history. In descriptions considered to refer megaesophagus, although dysphagia, the major symptom of this disease, is not recognized, typical manifestations of a irritating, inflammatory or ulcerative condition are identified, not affecting the esophagus but the stomach. In the description considered to refer to megacolon, the signs and symptoms suggest the diagnostic possibility of hemorrhoids and of the "achaque do bicho" itself, and do not recall the clinical picture of the chagasic megacolon in an absolute manner. On this basis, there is no reason to maintain the book "Noticias do que he o achaque do bicho" within the history of the digestive form of Chagas' disease. PMID- 9201323 TI - [Videofluoroscopy in the study of the oral and pharyngeal phases of deglutition]. PMID- 9201324 TI - [Swallowing defects determined by tracheostomy]. AB - We have studied the interference on swallowing of the skin-tracheal fixation determined by tracheostomy. We have analyzed this interference by videofluoroscopy. One hundred and twelve patients with complain of dysphagia have been studied by videofluoroscopy at the Hospital Universitario "Clementino Fraga Filho", Rio de Janeiro, RJ. Four have undertaken tracheostomy. One (female, 52 years) had already a metal cannula in the trachea, three others (two females and one male/40 to 66 years) exhibited a longitudinal anterior depressed scar, for more than five years. Two patients had also a neurological disease. In the videofluoroscopic observation it was used liquid medium (barium solution) and also solid and soft contrast media made of barium powder mixed with a bread dough. We have found a correlation between skin-tracheal fixation and swallowing defects. The skin-tracheal fixation interference occurs basically by opposition to the hyo-larynx free displacement. This limited displacement determines a small amplification of the laryngo-pharyngeal space, a restrictive opening of the pharingo-esophageal segment and also determines that the laringeous aditus remains near the pharingo-esophageal limit. We have observed swallowing defects in all the four tracheostomized patients. The observed defects relative to tracheostomy was the penetration of contrast medium in the airway. This penetration was cleared by forced expiration. In three patients the swallowing defects was clearly linked to tracheostomy. We can admit that the skin-tracheal fixation, without any other pathology, can determine dysphagia. In association with other diseases this fixation can increase the disturbance: On the other hand it can run unnoticed due to more evident pathology. The consequence of skin tracheal fixation is better determined by videofluroscopy. PMID- 9201325 TI - Diet of South Asians with inflammatory bowel disease. AB - The diet of South Asian patients with inflammatory bowel disease was studied using a postal questionnaire and compared with controls matched for age and ethnic group. Hindus with Crohn's disease are sweets significantly more often than either healthy controls (chi 2 = 13.0, P < 0.005) or Hindus who had ulcerative colitis (chi 2 = 9.8, P < 0.05). Hindus with ulcerative colitis were significantly less likely to drink milk than their matched controls (chi 2 = 7.4, P < 0.01). Hindu patients were also less likely to use spices and eat flour than controls (chi 2 = 12.5, P < 0.005). No difference was found in the diets of patients belonging to other religious groups and controls. This study again suggests that a high intake of refined carbohydrate is associated with Crohn's disease, while patients with ulcerative colitis have significantly altered their traditional diet. PMID- 9201326 TI - [Protective effect of reduction of pancreatic enzyme content on the pulmonary disease induced by acute pancreatitis]. AB - Lung injury develop in up to 50-70 percent of patients with acute pancreatitis. Cerulein in physiological doses reduces the rate mortality of pancreatitis by decreasing the enzyme content of the pancreas. In order to assess the effect of acute reduction of pancreatic enzyme content on the pancreatitis pulmonary injury, pancreatitis was induced in Wistar rats by intraductal injection of 5 per cent sodium taurocholate: group I, with pancreatitis; group II, pancreatitis after decreasing pancreatic enzyme content; group III, control group. Dye Evans blue was used to evaluate the lung injury. The pancreatitis pulmonary injury was smaller in group II than group I (P < 0.05) but it was similar to control group (group III). In conclusion, it is speculated that the reduction of the pancreatic enzyme content reduces the pancreatitis pulmonary injury by decreasing the quantity of pancreatic enzymes reaching the systemic circulation. PMID- 9201327 TI - [Endoscopic laser lithotripsy for difficult calculi after unsuccessful extracorporeal shock wave lithotripsy]. AB - INTRODUCTION: More than 97% of common bile duct stones can be successfully managed by endoscopic papillotomy, mechanical lithotripsy and extracorporeal shock-wave lithotripsy. In this study, we evaluate the role of laser lithotripsy after failure of the above mentioned endoscopic methods. PATIENTS AND METHODS: Eighteen patients (15F, 3M; median age = 69 (28-83) years) were treated by endoscopic laser lithotripsy after ESWL failure. We employed a Rhodamine-6 G laser with a stone-tissue recognizing system. The laser fibers were cholangioscopically (direct vision) or blindly (under plain fluoroscopic control) placed. RESULTS: Seventeen patients were treated endoscopically and one was successfully managed percutaneously after failure of the transpapillary approach. Fourteen (78%) were stone-free after a mean of 1.56 laser therapy sessions alone. Two additional patients were successfully managed after partial fragmentation with combined treatment (mechanical lithotripsy: n = 1, electrohydraulic lithotripsy: n = 1). Overall, 89% of the patients were freed from their calculi. Cholangitis occurred once and the mortality was zero. CONCLUSIONS: We conclude that laser lithotripsy is an effective and safe treatment alternative in a highly selected patient population with difficult bile duct stones and considerable surgical risk. PMID- 9201328 TI - [Ultrasonically guided needle biopsy in malignant focal solid nodules of the liver]. AB - In the most of the cases, the diagnosis of focal solid hepatic lesions are performed by ultrasonography, computed tomography, magnetic resonance and hepatic angiography imagig. However, the distinction between benign and malignant neoplasias, sometimes is made after liver biopsy. This report is about 32 of these lesions, diagnosed after guided liver biopsy by ultrasonography. The efficacy of this propedeutic method, minimally invasive, is defined emphasizing that there are no mortality and low levels of morbidity. PMID- 9201329 TI - [Microsurgical pancreatoduodenal transplantation in rats. Technique and results following 12 years of investigation]. AB - In this study we present the technical details, adaptations and modifications of the original procedure of pancreaticoduodenal transplantation in rats described by Lee et al. in 1972. We also present the results and technical failures observed in a follow-up of 12 years. From March, 1982 to December, 1994, we performed in the Laboratory of Surgical Technique and Experimental Surgery of Faculty of Medicine, Botucatu-UNESP, Brazil, 665 duodenopancreatectomies in donor rats and 592 surgeries for revascularization of the pancreatic graft in recipient animals. The observed percentage of technical failures in donor rats was 11% due to bleeding and/or vascular complications, irregular flushing of the graft with saline and respiratory insufficiency. In recipients of grafts, we observed a percentage of technical failures of 22.5% due to porto-caval thrombosis, vascular bleeding, pancreatitis and graft ischemia. In both surgeries, the successful results are directly related to the technical performance of the surgeon and the cares in the postoperative period. PMID- 9201330 TI - [Glucagonoma: case report and literature review]. AB - Glucagonoma is a neuroendocrine tumor of pancreatic alpha cells manifested by necrolytic migratory erythema, hyperglucagonemia, glucose intolerance, weight loss, anemia and hypopaminoacidemia. We report a case of glucagonoma in a 38 years-old patient diagnosed by the presence of a pancreatic tumor, liver metastasis, weight loss, glucose intolerance, necrolytic migratory erythema, hyperglucagonemia (1400 pg/ml; normal < 200 pg/ml) and histologic demonstration of glucagon and neurospecific enolase by immunocytochemical reaction. Actual therapeutic of glucagonoma includes surgery, chemotherapy, somatostatin or octreotide for control of the symptoms, and more recently alpha-interferon was suggested. PMID- 9201331 TI - [Biological activity of fish oil]. AB - Omega-3 fatty acids (n-3) found specially on fish oil are represented by the acid alfa-linolenic, eicosapeniaenoic and docosahexaenoic. After 72 hours of n-3 fatty acids intake there are changes on membrane composition and decrease of synthesis of prostaglandin (PG), leucotriens (LT) and thromboxanes (TX) of the 2 and 4 series production and substitution for prostaglandin, thromboxanes and leucotriens of 3 and 5 series respectively. These alterations can modulate the inflammatory response in some diseases. N-3 fatty acid have been used in cardiology on hypercholesterolemy and hypertension control and on immunologic diseases as psoriasis, rheumatoid arthritis and Crohn disease. Experimentally the n-3 fatty acid inhibit tumor growth and metastasis. The authors consider the biophysiological actions of n-3 fatty acids and discuss the results of its use on clinical results. PMID- 9201332 TI - Report of two cases of children with Budd-Chiari syndrome successfully treated with streptokinase. AB - Two children with Budd-Chiari syndrome were successfully submitted to thrombolytic therapy. This study suggests that streptokinase is safe and effective in the treatment of this syndrome and should be considered as primary treatment in case of early diagnosed acute disease in view of the poor prognosis and the aggressiveness of surgical treatment currently available. PMID- 9201333 TI - [Meningiomas. Epidemiological and anatomopathological study of 340 cases]. AB - The authors have retrospectively reviewed all tumors of central nervous system (CNS) operated at the most important neurosurgery hospitals of Curitiba in a 5 year period (1990-1994) and found 304 (22.4%) cases of meningioma. Age mean of the patients was 48.5 years, with a range of 3 to 90 years. A marked female preponderance (68.7%) was noted. The most common tumor location was brain (n = 280) and the remaining cases occurred in spinal cord (n = 10), cerebellum (n = 9) and cranial nerves (n = 5). Histologically, there were 294 (96.7%) meningiomas of the classical type, six malignant or anaplastic, three atypical and one papillary. Two hundred and sixty seven classical meningiomas were from the meningotelial subtype, ten psamomatousos, five fibroblastic, five microcystic, five transicional and two angiomatous. The authors conclude that meningiomas are one of the most common group of primary neoplasias of CNS and the definition of malignancy in those tumors is beset by frequent discordance between histologic and biologic features. PMID- 9201334 TI - [Compulsory notification of cysticercosis in Ribeirao Preto-SP, Brazil]. AB - Cysticercosis is a severe public health problem in several regions of Asia, Africa and Latin America. Epidemiologic studies based on the frequency of cases observed in specialized neurology, neurosurgery and computed tomography services, at autopsy and in seroepidemiologic studies do not permit the determination of the true prevalence of the disease in the population. The objective of the present study was to investigate the prevalence of cysticercosis by compulsory notification. The coefficient of prevalence was 54 cases/100,000 inhabitants in the municipality of Ribeirao Preto. The results also indicated that cysticercosis is not under control in our region since 21% of cases presented the active form of the disease. Compulsory notification proved to be a valuable resource for the epidemiologic study of cysticercosis, also permitting the mapping of more affected areas for a better direction of prevention strategies. PMID- 9201335 TI - Neurocysticercosis in Paraiba, Northeast Brazil. An endemic area? AB - Neurocysticercosis is the central nervous system infestation by Cysticercus cellulosae, the larval form of Taenia solium. It is related to poor hygiene habits and sanitation; although Northeast is poorest Region of Brazil, it has been always stated as a non-endemic area. After the installation of computed tomography (CT) service, the incidence of neurocysticercosis began to raise in neurology services in Campina Grande PB, a city where people from the interior Paraiba can find specialized medical facilities. We analyse 5,883 CT record of the TomoHPI Computed Tomography Service from August 1993 to December 1995, observing 1.02% suggestive neurocysticercosis cases and classified them according to sex and age, procedence and socioeconomic condition. Distribution of cases according to age is homogeneous until the age of 50 (mean: 28.36 years old). Men and women are equally affected. Urban areas inhabitants represented 83.33%. Residents of Campina Grande represented 48.33% and 48.34% were residents of cities around Campina Grande (until 50 Km around) and other cities of Paraiba State. Fifty-eight patients were dependent to public health care system. We conclude that neurocysticercosis seems to be endemic in Paraiba State, demanding a more detailed study to determine its incidence/prevalence. PMID- 9201336 TI - [Clinical and laboratory characteristics of bacterial meningitis in children]. AB - Data from the records of 528 children under 15 years old with diagnosis of acute bacterial meningitis, admitted at the Hospital Couto Maia between 1990 and 1992 were analyzed. Bacterial meningitis was more frequent in children under the age of 1 year (37.8%). The most common etiologic agent was H. influenzae (42.2%). The global lethality was 20.9%. Individual predictors of poor outcome were: absence of the "classic triad", CSF cell count under 1000/mm3, age under 2 years, presence of seizures, depressed sensorium, and S. pneumoniae as causal agent. PMID- 9201337 TI - Creutzfeldt-Jakob disease. A survey of 14 patients. AB - Creutzfeldt-Jakob disease (CJD) is a transmissible disease of the nervous system causatively related to the presence of an abnormal prion protein, with dementia, myoclonic jerks, and periodic EEG activity. Fourteen patients (7 females and 7 males) ranging from 26 to 76 years of age (median 59 years) were evaluated between 1974 and 1995 at the Neurologic Clinic of Sao Paulo University School of Medicine. The average duration of the disease was 12 months (3.5-34 months). Early clinical findings were: behaviour changes in 7 patients, dementia in 4, visual disturbances in 4, vertigo in 2, tremor in 9, and dystonia in one. Advanced symptoms were dementia and myoclonus in all patients. Pyramidal tract dysfunction was found in 6, cerebellar ataxia in 2, seizures in 3, nystagmus and vertigo in 4, and peripheral nervous system involvement in 2. Atypical clinical forms were found in 5 patients. Periodic EEG activity was found in 10 patients. Cerebrospinal fluid evaluation showed pleocytosis in 1 patient, higher protein content in 2, and higher gamma globulin level in 2. In 10 patients anatomopathological evidence in the central nervous system confirmed the clinical diagnosis by presenting with status spongiosus. All except one patient presented with the sporadic form of the disease. PMID- 9201338 TI - [Antiphospholipid antibodies in 66 patients with cerebral infarction between 15 and 40 years old]. AB - The antiphospholipid antibodies (aPLs) are a heterogeneous group of immunoglobulins that have been related with alterations in blood coagulability in recent years. Patients with elevated titers of these antibodies have a high probability to develop thrombotic events, including cerebral infarct (CI). The tests currently used to detect these antibodies are the lupus anticoagulant and ELISA for anticardiolipin antibodies which have a larger proportion of positivity among young patients with CI. In our study we tested 66 patients with cerebral infarcts whose ages ranged from 15 to 40 years for the presence of lupus anticoagulant and anticardiolipin antibodies. The results showed that eleven (16.65%) patients were positive for aPLs and three (4.55%) of them fulfilled the diagnostic criteria for primary antiphospholipid syndrome. These data point out to the importance of investigating aPLs in young patients with CI and its high prevalence in this group compared with healthy population. PMID- 9201339 TI - [Cerebral infarctions in young patients related to deficiency of natural anticoagulants. Protein C and protein S]. AB - The possible etiologies of cerebral infarcts (CI) in young patients differ from those in the older stroke population. Recently, deficiencies of fibrinolytic factors emerged as an important etiology of stroke in the young population. The literature has emphasized the diagnosis of such conditions especially in stroke cases of undetermined etiology and with history of recurrent thrombosis. We carried out a research on the serum level of protein C, protein S, and antithrombin III in young patients, between 15 and 40 years, with CI of undetermined cause after application of a standard protocol. Three patients had low levels of these proteins; two had protein C deficiency, and one protein S deficiency. None of them had antithrombin III deficiency. We conclude that systematic investigation of such causes of cerebral infarct will reduce the group of undetermined strokes, and will open the possibility of prophylactic treatment in a non-negligible group of patients. PMID- 9201340 TI - [Congenital myotonia. Report of 7 patients]. AB - Myotonia is the phenomenon of decrease of muscular relaxation rate, after either a contraction or a mechanical or electrical stimulus. Congenital myotonias are hereditary affections and do not present muscular dystrophy. The current trend is to group them as ionic channels diseases, together with the periodic paralysis. The authors accompanied the cases of seven patients, six males and one female, with ages ranging from 16 to 48 years (average 27 years) and onset of symptoms between 1 and 10 years (average 5 years). These patients presented a myotonic phenomenon unleashed by intensive contraction and global muscular hypertrophy. Three patients were diagnosed as cases of Becker type generalized myotonia because they presented a recessive autosomic heredity and/or transient episodes of muscular weakness. Two patients fitted the description of Thomsen congenital myotonia, with a pattern of dominating autosomic heredity and/or absence of weakness episodes or worsening factors for their condition. Two patients presented fluctuating myotonia, which because worse in cold weather or at potassium intake. The clinical diagnosis was confirmed through complementary tests (electroneuromyography, muscle biopsy and DNA study). Each of the patients made use of different drugs, in the search of optimal lessening of their myotonia. There were five reports of amelioration with the use of diphenilhydantoine; one report with the use of carbamazepine; three reports with the use of acetazolamide; one report with the use of a calcium channel blocker; one report with the use of a beta-adrenergic; one report with the use of thiazide; and none with the use of quinidine/procainamide. PMID- 9201341 TI - Evaluation of the respiratory function in myasthenia gravis. An important tool for clinical feature and diagnosis of the disease. AB - Myasthenic gravis may affect both inspiratory and expiratory muscles. Respiratory involvement occurred in almost all patients with myasthenia gravis in all clinical forms of the disease: 332 lung function tests done in 324 myasthenic patients without respiratory symptoms (age 34.6 +/- 18.3 years) were examined. Lung volumes analysis showed that all the patients of both sexes with generalized or ocular myasthenia gravis showed "myasthenic pattern". Male patients with "ocular" form only presented the "myasthenic pattern" with lung impairment and had, from the lung function point of view, a more benign behaviour. Female patients with the "ocular" form exhibited a behaviour of respiratory variables similar to that of the generalized form. It was not observed modification of the variables that suggested obstruction of the higher airways. The "myasthenic pattern" was rarely observed in other neuromuscular diseases, except in patients with laryngeal stenosis. PMID- 9201342 TI - [Osteochondromas of the spine. A diagnosis to consider in spinal cord compression syndromes]. AB - The authors reviewed 312 cases of solitary and multiple osteochondromas seen in SARAH Hospital for the Locomotor System during a period of 13 years-from 1982 to 1994. They selected six cases of patients with osteochondromas of the spine, corresponding to 1.92% of the total number of diagnosed cases of this entity. The selected cases were submitted to roentgenographic examination that comprised plain roentgenograms, myelography, computerized tomography and, in one of them, magnetic resonance imaging. They were submitted to decompressive surgical procedures (including laminectomy) with exeresis of the lesions and posterior histopathological examination which confirmed the initial diagnostic hypothesis. This study also includes a review of the possible mechanisms implicated in the pathogenesis of the disease. PMID- 9201343 TI - [Temporal lobe epilepsy. Surgical treatment]. AB - The authors report the surgical management of 32 patients with medically intractable seizures. In all cases the epileptiform focus present in the temporal region was demonstrated by electroencephalography. Our report was made up of 14 male patients and 18 female patients. Their ages ranged from 9 to 62 years. The material was divided into two groups. The first, with eighteen patients with cerebral lesion (like gliomas, arteriovenous malformation, epidermoid tumor) demonstrated on the CT scan and MR imaging underwent to lesion resection: in some cases with adjacent irritative area (guided by electrocorticography) out of eloquent zone, the removal of this irritative area was done. The second, with fourteen patients without cerebral expansive lesion; the MR imaging showed mesial temporal sclerosis in eight cases; all the patients of this group underwent to temporal lobectomy; the histopathologic exam showed temporal sclerosis in nine cases and normal brain in five. The postoperative follow-up showed better results in the cases with expansive cerebral lesion (83.4% seizure free) than the cases without that lesion (71.4% seizure free). PMID- 9201344 TI - The hyperactive child and the body. A clinical study on the origin of hyperactivity in children. AB - A group of 22 hyperactive children from 7 to 12 years of age was selected among 38 out-patients registered at Hospital do Servidor Publico de Sao Paulo (Civil Servant Hospital of the State of Sao Paulo). Their psychiatric evaluation was negative, the neurological examination showed "psychomotor syndrome", and psychological evaluation revealed disorders related with Ego maturation in all cases. Although all children were referred to psychotherapy, only thirteen underwent individual sessions once a week for an uninterrupted period of up to one year. Neither diets nor medicines were prescribed. After six months and one year of treatment, the children were reevaluated. They showed improved school performance, reduced hyperactivity, and better internal psychic organization. These results are considered as undeniable evidence of the psychodynamic origin of hyperactivity syndrome in children, when no definite neurologic or psychiatric diseases are demonstrated. PMID- 9201345 TI - [Which patients does the neurologist assist? Basis for a curriculum in neurology]. AB - OBJECTIVE: To present the most frequent diagnosis of patients referred to a neurologist and to discuss the importance of this finding for the definition of the curriculum in Neurology. BACKGROUND: The development of subspecialties of Neurology is interfering in the definition of what should be taught to train a physician or a neurologist. The knowledge of which are the most common neurological diseases may contribute to construct these curricula. METHOD: The initial diagnosis in 1815 outpatients referred to the neurologic service of an university-affiliated public hospital in Sao Paulo, Brazil, were analyzed. RESULTS: The most common diagnosis, in decreasing order of frequency were: headache, epilepsy, mental disorders, cerebrovascular disease, head injury, polyneuropathy, vestibular syndrome, spastic crural paraparesis, extrapyramidal syndrome, dementia, intracranial hypertension and facial palsy. CONCLUSION: The importance of the subspecialties in the curriculum should be related to the frequency of the neurologic diseases in the community. PMID- 9201346 TI - [Micro-mesoscopic study of the lateral wall of the human cavernous sinus]. AB - The authors studied the structures of human cavernous sinus in its interior as well as on the lateral wall, utilizing thick, frontal, sequential sections. They show the significance of this wall, frequently used as surgical accessway to diseases encountered within this venous structure of the dura-mater. PMID- 9201347 TI - [Acute hepatic failure associated with valproic acid in children. Report of 3 cases]. AB - We report the cases of three epileptic children who developed hepatotoxicity induced by valproic acid. Two patients had developmental delay. Including the one who died, all patients were receiving polytherapy (carbamazepine in two and phenobarbital in one). The patients age ranged from 2 years and 8 months to 5 years and 1 month. The onset of hepatic complications occurred within 6 months of valproate therapy in two patients and 12 months in one. All patients developed the classical clinical signs of hepatotoxicity. Vomiting, edema and jaundice were the initial symptoms. Fever occurred in two patients. The serum levels of glutamic oxaloacetic transaminase were mildly elevated with a maximum of 194 IU. The bilirubin levels ranged from 5.5 to 19.8 mg%. Two patients recovered clinically and showed normalization of the laboratory abnormalities and one had fatal course. The hepatotoxicity must be considered as a side effect of valproic acid mainly in children under two years age, with polytherapy regimen and neurologic damage. The hepatic insufficiency can be reversible. PMID- 9201349 TI - [Cerebral vasculopathy in the primary antiphospholipid antibody syndrome. Report of 2 cases]. AB - The antiphospholipid antibodies are associated with a large number of neurologic syndromes, cerebral infarct (CI) being the most common of them. In these cases the pathogenesis of the CI is poorly understood and remains controversial; however, the existence of a vasculopathy is indubitable. We report the cases of two young patients with CI and diagnosis of primary antiphospholipid syndrome who were submitted to cerebral angiograms, and one of them to necropsy. In one case the angiographic findings were similar to those of vasculitis in intracranial vessels. In the other case we observed obstruction in internal carotid artery at the angiography that looked like thrombosis in situ; at necropsy we found non atherosclerotic obstruction in coronary arteries. In summary, is the primary lesion vasculitis, thrombosis, or both? These cases illustrate the discussion and demonstrate that vasculitic mechanisms may be involved in the vasculopathy of primary antiphospholipid syndrome even though thrombosis occur more frequently. PMID- 9201348 TI - [Familial cavernous angioma. Report in 3 generations]. AB - Cavernous angioma is a vascular malformation that affect 0.5 to 0.7% of the population making up 8 to 15% of cerebrovascular malformations. It is the second vascular malformation in frequency of the central nervous system, supplanted only by classic arteriovenous malformation. It may occur in two forms: a sporadic form characterized by isolated lesions: and a familial form characterized by multiple lesions with an autosomal dominant mode of inheritance with high penetrance and varied expressivity in the proportion M1:F1. Symptoms related to cavernous angioma are seizures, headache or progressive neurologic deficit. The authors present a Chinese family with familial cavernous angioma. Manifestations of the disease occurred in three generations affecting only females. Clinical, neuroimage, pathological, natural course and genetical aspects of the disease are discussed. PMID- 9201350 TI - The "one-and-a-half" syndrome. Case report. PMID- 9201352 TI - [Giant intracranial aneurysm in a 9 year-old child. Case report]. AB - The case of a 9-year-old girl with intracranial hemorrhage due to a giant aneurysm of distal middle cerebral artery is reported. Cerebral aneurysms are rare in children, particularly in the first decade of life. Aneurysms in childhood occur more frequently in peripheral branches and they are more often giant in size. Clinical features and pathogenesis of this lesion are discussed. PMID- 9201351 TI - [Cruciate hemiplegia associated with basilar impression, Arnold-Chiari malformation and syringomyelia. Case report]. AB - The authors report a case of cruciate hemiplegia associated with basilar impression, Chiari malformation and syringomyelia. The neuroanatomical controversy, the surgical treatment and the good outcome of the patient are discussed. PMID- 9201353 TI - [Acute schistosomiasis with brain involvement. Case report]. AB - A case of acute schistosomiasis with magnetic resonance images (MRI) of the brain suggestive of demyelinating lesions, pyramidal disorder in the lower limbs and normal cerebrospinal fluid is presented. Diagnosis could be established by detection of antibodies on blood and cerebrospinal fluid. Schistosoma mansoni involves the spinal cord more often than the brain. Praziquantel associated to prednisolone was effective in this case. There are few reports of brain involvement with S. mansoni, but its prevalence is probably greater. Due to the paucity of studies, its pathophysiology and therapeutics remain to be better clarified. The immune and MRI aspects are emphasized. PMID- 9201354 TI - [Idiopathic lumbosacral plexus neuropathy in child. Case report]. AB - We describe a lumbosacral plexus neuropathy case in childhood in which detailed investigation, including electromyography and magnetic resonance imaging, was normal. Muscle biopsy showed mild denervation. No underlying condition was detected. The patient presented with pain, weakness and light atrophy in left lower limb, reduced reflex at the ankle, loss of the quadriceps reflex and paresthesy in involved limb. Recovery after one year was almost complete, with persistent slight weakness and atrophy. PMID- 9201355 TI - [Dementia in Parkinson's disease. Critical evaluation of the literature]. AB - In the last 30 years, Parkinson's disease has been object of great progress. The majority of patients reaches a longer life with quality because of the modern therapeutic approach. However, dementia that can occur in the evolutive process, has its neuropathology not completely defined until now. There are lesions in the basal ganglia, in the ventral area of the mesencephalic tegmentum in the thalamus, in the substantia nigra and in the frontal cortex. The presence of Lewy bodies in the cortex is associated with dementia, in the same way that the anatomopathological features of Alzheimer's disease, in many cases. Dementia should have a multifactorial basis. Different types of neurotransmitters, like serotonin, acetylcholine and dopamine, or even hormones, like cortisol, may be altered in a great number of demented parkinsonians. Depression, found in up to 40% of patients, have been related as a risk factor for dementia, present approximately in 25% of cases. Studies in this area are still conflicting, with some confirming the relation among depression, cortical atrophy, hypercortisolemia and Parkinson's disease. Neuropsychologic studies show that the dementia in Parkinson's disease is of subcortical type. It is also known that parkinsonians, even those without cognitive deficiencies clinically significant, present deficits if submitted to more detailed neuropsychological tests. It is assumed, so, that cognitive impairments are intrinsic to the disease, varying its expression among patients. Dementia shall be diagnosed based on the criteria established in the diagnostic and statistical manual of mental disorders of the American Psychiatry Association, as well as computed tomography and magnetic resonance. For treatment, parkinsonian dementia does not recognize efficacious agents until now. PMID- 9201356 TI - [Early onset Parkinson's disease. Critical review of the literature]. AB - Since its original description Parkinson's disease has been considered as a clinical condition which affects older people. Nonetheless, since late in the last century, cases starting in very young age have been described. A great controversy has arisen concerning the real pathology in these cases and, consequently, how should they be named. Early or young onset parkinsonism, early or young onset Parkinson's disease, juvenile parkinsonism, all these terms have been used indistinguishable. There have been few pathological descriptions in early onset parkinsonism. Some show striking differences from the cases of older patients but others are very similar to what has been considered classical Parkinson's disease. Younger starting age usually corresponds to greater possibility of other family members being affected. Dyskinesias and fluctuations due to chronic levodopa treatment are an early and almost invariable complication in the course of young patients. Comments on several aspects based on an extensive literature review are presented. PMID- 9201357 TI - [From death concepts to brain death diagnostic criteria]. AB - The authors present considerations about death and brain death concepts, as well the legal aspects for its diagnosis in Brazil. They also present the UNICAMP Protocol for the Diagnosis of Brain Death, revised and according with the current law, with standard techniques for the diagnostic exam. They emphasize the importance of a mature ethical position for this frequent and challenging situation. PMID- 9201358 TI - Anisakis, anisakidosis, and allergy to Anisakis. PMID- 9201359 TI - Bronchial epithelial cells in asthma. PMID- 9201360 TI - European Academy of Allergology and Clinical Immunology (EAACI) guidelines for continuing medical education. Essentials for accreditation, standards for commercial support, and system of credits. PMID- 9201361 TI - Physical urticaria: classification and diagnostic guidelines. An EAACI position paper. PMID- 9201362 TI - Cross-reactivity between IgE-binding proteins from Anisakis, German cockroach, and chironomids. AB - Anisakis simplex larvae parasitize animals used as seafood and can produce a specific immune response in man. The ingestion of seafood contaminated with stage three of A. simplex larvae can induce a specific IgE response with clinical symptoms, usually urticaria, even if the fish is cooked before ingestion and the invasive infestation power destroyed by heating. Our preliminary studies showed a strong association of A. simplex sensitization with Ascaris lumbricoides, Daphnia, chironomid spp., Atlantic shrimp (Pandalus borealis), and German cockroach (Blattella germanica). We conducted the cross-reactivity study with cockroach, a ubiquitous insect, and Chironomidae (red mosquito larvae), a work related allergen, without any possibility of Anisakis contamination. Serum samples were collected from 60 pediatric patients, with serum specific IgE to A. simplex. Both specific-IgE and immunoblot-inhibition studies, with a serum pool from 18 patients, were performed to determine whether the association of sensitizations to nematodes and arthropods was due to immunologic cross reactivity. In addition, serum samples from 21 of 60 patients who showed also sensitization to German cockroach were used for individual immunoblot studies. In the serum pool, dose-dependent inhibition of B. germanica and Chironomus spp, was observed after preincubation with the A. simplex extract. Immunoblot of Anisakis, inhibited with Chironomus and German cockroach, yielded a partial blot inhibition but mainly on bands below 41 kDa. Blot inhibition of German cockroach and Chironomus with Anisakis was dose related. The band patterns in individual blots were heterogeneous, but most of them had bands of 30-43 kDa. None of these sera recognized allergens in the 14-kDa area. In our study, CAP-inhibition and immunoblot-inhibition analysis of Anisakis showed that several IgE-binding components could be shared by the three allergens. PMID- 9201364 TI - Physical capacity and dyspnea in patients with asthma-like symptoms but negative asthma tests. AB - Ten female patients with asthma-like symptoms but negative asthma tests (study group) were compared with 10 female asthmatics in an exercise test with and without pretreatment with beta 2-stimulants. The aim was to determine whether the asthma-like symptoms in the patients of the study group could be explained by bronchoconstriction, circulatory abnormalities, or physical unfitness when provoked physically, and whether the exercise test could be used to distinguish these patients from asthmatics. Without pretreatment, the asthma group reacted with bronchoconstriction, as indicated by postexercise systematic changes in PEFR, FEV1, FVC, and SaO2, which were not seen in the study group. The groups differed in the ratings of "difficulty in getting air", as only the asthma group had significantly lower ratings when pretreated. The study group's mean test performance was 94 W; the asthma group's was 106 W. The low performance was not explained by disturbances in heart rate, electrocardiogram, or blood pressure or physical unfitness. The exercise test was found to distinguish between the groups, especially for bronchoconstriction, oxygen saturation, and ratings of dyspnea. It could be used complementary to lung function tests to eliminate bronchoconstriction, circulatory abnormalities, and physical unfitness as a cause of the asthma-like symptoms. PMID- 9201363 TI - Intercellular adhesion molecule-1 on cultured human epithelial cell lines: influence of proinflammatory cytokines. AB - The expression of intercellular adhesion molecule-1 (CD54 or ICAM-1) on epithelial cells during acute or chronic inflammation may favor the interaction between epithelial cells and leukocytes expressing the natural ligands of ICAM-1, LFA-1 (CD11a/CD18), and Mac-1 (CD11b/CD18). We have evaluated in vitro the expression of ICAM-1 by a conjunctival (WK) and an intestinal (I407) human continuous epithelial cell line. Cells were cultured for 24 h in the presence or absence of IFN-gamma, TNF-alpha, IL-1 beta, IL-4, IL-6, IL-8, IL-10, and TGF-beta 1. Both epithelial cell lines showed a constitutive expression of ICAM-1. IFN gamma at 500 U/ml and TNF-alpha at 200 ng/ml upregulated ICAM-1 expression; IL-1 beta at 100 pg/ml upregulated ICAM-1 on WK cells only. Cells cultured in the presence of both IFN-gamma and TNF-alpha exhibited a mean fluorescence intensity far greater than those cultured with IFN-gamma or TNF-alpha alone. I407 and WK cells were able to release soluble ICAM-1. IFN-gamma and TNF-alpha enhanced the release of sICAM-1. IL-4, IL-6, IL-8, IL-10, and TGF-beta 1 did not affect either ICAM-1 expression or sICAM-1 release. In conclusion, continuously cultured human epithelial cells may express ICAM-1 on their surface and release it in culture medium. These phenomena are upregulated by proinflammatory cytokines. PMID- 9201365 TI - Comparison of direct immunostaining and electroimmunoassay for analysis of airborne grass-pollen antigens. AB - Sensitive pollen-allergic patients have been reported to show allergic symptoms not only during the pollen release of allergenic plants but also both before and after the pollen season. Symptoms before the season are evidently provoked by small-sized particles originating partly from developing pollen grains, partly from other plant parts. After the pollen season, antigenic material settles on various surfaces, which thus form a new source of allergenic material. Measuring the allergen concentrations in indoor and outdoor environments demands an effective sampling method and a rapid and sensitive immunochemical analysis, especially for particles of small-sized fractions which are not detected in microscopic analyses. The efficiency of an ELISA and an immunochemical staining method was tested with monoclonal IgG against Phl p 5, the main grass allergen. The Burkard trap and MPC impactor (Marple personal cascade impactor with six stage particle size fractionation) were compared. The sampling was carried out in southwestern Finland in the summer of 1994. The number of grass-pollen antigen spots greatly exceeded the simultaneous pollen count, indicating considerable antigen activity outside the pollen grains. The counts were especially high in small-sized fractions after the pollen season, when hardly any airborne pollen was found. Spots and pollen divided according to size were highly intercorrelated, indicating that the threshold values used were appropriate. PMID- 9201366 TI - Quantitative measurement of extracellular histamine concentrations in intact human skin in vivo by the microdialysis technique: methodological aspects. AB - Calculation of recovery is needed in microdialysis studied to calculate absolute concentrations of compounds in the extracellular water space. The purposes of this study were to determine the extracellular concentration of histamine in intact human skin in vivo and to study the validity of absolute histamine measurements during allergic skin reactions. A skin microdialysis technique and two calibration techniques, the no net flux method and the flow rate method, were used to quantify histamine concentrations in resting skin. To validate these techniques, skin glucose concentrations were analysed as well. In addition, the influence of vasodilation and plasma extravasation on recovery was followed after intradermal injection of codeine, a mast-cell secretagogue. As expected, both calibration methods estimated skin glucose concentrations to be identical with venous blood glucose concentrations. However, skin histamine levels could not be calculated by the no net method, because the data did not meet the theoretic assumptions of this method. In contrast, histamine data fitted theoretically with the flow rate method, and skin histamine concentrations of 18.8 +/- 2.8 nM were found to be significantly greater than plasma histamine concentrations of 4.3 +/- 0.7 nM. Within minutes after intradermal injection of codeine, recovery increased significantly in a dose-dependent fashion. Vasodilation per se did not influence recovery. In conclusion, absolute assessment of skin histamine concentrations can be made by microdialysis by the flow rate method. The validity of such an estimate and the theoretic prerequisites for the calculations are discussed. Quantitative measurement of skin histamine levels during allergic reactions cannot be performed since recovery is altered by plasma extravasation after skin challenge. PMID- 9201367 TI - Allergy to the ornamental indoor green plant Tradescantia (Albifloxia). AB - We report on a 32-year-old atopic female office employee with a moderate tree pollinosis who also suffered from indoor-related perennial rhinoconjunctivitis. Once when she repotted her two ornamental nonflowering green plants of the genus Tradescantia (synonym; Albifloxia; family Commelinaceae), she immediately experienced itching of the face, throat, and conjunctiva; swelling of the lips; and dyspnea and wheezing. Skin prick tests with the leaves of Tradescantia (T. albifloxia and T. fluminensis) (Ta and Tf) were strongly positive as was the specific IgE to Ta leaves extract. On RAST inhibition studies, no cross reactivity was found between Ta and Ficus benjamina (weeping fig), a nonflowering green plant, which produces, in its milky sap, an important respiratory allergen. Green plants should be considered potential indoor allergens and tested in plant keepers referred for allergologic investigation. PMID- 9201368 TI - Relevance of pollen-specific IgE levels to the development of Apiaceae hypersensitivity in patients with birch pollen allergy. AB - A large clinical/serologic study was carried out to determine the prevalence of Apiaceae (carrot, celery, and fennel) hypersensitivity in patients with birch pollen allergy, and to investigate its relationship with apple and hazelnut allergy and with birch pollen-specific IgE levels. A total of 196 birch pollen hypersensitive patients with oral allergy syndrome (OAS) caused by different vegetable foods were examined in the cross-sectional part of the study. Of this total, 195 patients had apple and/or hazelnut allergy, and 103 had Apiaceae sensitivity; only one patient had Apiaceae allergy alone. Apiaceae-positive patients showed significantly higher birch pollen-specific IgE levels than negative ones (median 13 vs 7 AU/ml; P < 0.0001). The prospective part of the study was performed on 103 birch pollen-hypersensitive patients who were OAS-free at the time of the first visit and were periodically followed-up for OAS. Patients who developed Apiaceae sensitivity showed much higher birch-specific IgE levels than patients who developed apple/hazelnut allergy only (median 15.5 vs 8.5 AU/ml; P < 0.05), whereas those who remained OAS-free showed the lowest specific IgE levels (median 5 AU/ml). This study suggests that most Apiaceae determinants cross-react with apple or hazelnut determinants, whereas only some apple or hazelnut determinants cross-react with Apiaceae-allergenic determinants; moreover, it shows that birch-specific IgE levels heavily influence the onset of OAS as a whole, and probably play a critical role in the development of allergies to distinct vegetable foods as well. PMID- 9201369 TI - Periodate treatment of Anisakis simplex allergens. AB - Anaphylactic reactions after parasitized fish consumption are mediated by an IgE response. However, positive skin tests and specific IgE can also be found in many asymptomatic subjects who recognize a single medium-mol.-wt. antigen by IgE immunoblot. The study aimed to find out whether this unspecificity was due to the carbohydrate moieties of parasite antigens. Sixty-two patients with suspected parasite allergy, 51 blood donors, 18 bakers, and 38 atopic patients were studied by blotting. Parasite proteins were treated with periodate. Several selected sera were inhibited with a crude wheat extract and fungal amylase. Twelve patients (19%), eight donors (16%), six bakers (33%), and one atopic patient (3%) recognized a single medium-mol.-wt. band in blotting and should be considered false-positive. This band was periodate-sensitive, but specific IgE to this allergen could not be inhibited by a wheat extract nor by fungal amylase and was clinically irrelevant. Diagnosis of Anisakis simplex hypersensitivity by skin tests and/ or specific IgE values should always be confirmed by specific IgE immunoblotting in order to detect the presence of clinically unrelated antibodies directed to periodate-sensitive allergens. These allergens are probably not a carbohydrate moiety of a parasite glycoprotein. PMID- 9201370 TI - Prevalence of childhood asthma in Istanbul, Turkey. AB - In order to determine the asthma prevalence in 6-12-year-old schoolchildren in Istanbul, we issued 2350 questionnaires, according to ISAAC criteria, in six randomly selected city primary schools to be completed at home by parents. A total of 2232 of the questionnaires were completed, an overall response rate of 94.9%, and 2216 questionnaires were taken into consideration. The prevalence of asthma was found to be 9.8% and wheezing 15.1%. To investigate the effect of socioeconomic status on the prevalence of asthma, we evaluated the heating system at home, the place of residence, the educational levels of the mother and father, the number of people living in the house, the sharing of bedrooms, and the annual family income. In conclusion, the prevalence of childhood asthma was not affected by any of these factors. Atopic family history, food allergy, eczema, and frequent otitis media and sinusitis attacks were evaluated and found to be significant in asthma prevalence. PMID- 9201371 TI - Anisakis simplex, a relevant etiologic factor in acute urticaria. AB - Anisakis simplex, a parasite of fish and cephalopods, can induce IgE-mediated reactions. This study aimed to determine the etiologic role of A. simplex in patients affected by urticaria/angioedema (AE) or anaphylaxis. We studied 100 adult subjects suffering acute episodes of urticaria/AE, by anamnesis, prick tests with A. simplex and fish-mix extracts, and total and specific IgE to both A. simplex and cod. The following criteria of A. simplex allergy were considered: 1) urticaria/AE within 6 h after fish ingestion; 2) specific IgE to A. simplex; 3) positive prick test to A. simplex extract; 4) exclusion of other suspected causes. Double-blind, placebo-controlled food challenge was not carried out because ethical considerations forbid challenge with a parasite. Specific IgE to A. simplex (> 0.7 kU/l) was found in 22 subjects, but only eight were diagnosed as having A. simplex allergy. Other allergens were involved in 37 patients, and 55 cases were considered idiopathic. Specific IgE to fish (> 0.7 kU/l) was found in two patients, but only one was diagnosed as having fish allergy. We concluded that A. simplex is an important etiologic factor in acute urticaria. We suggest that it should be considered in cases of urticaria/AE or anaphylaxis, especially after fish ingestion. PMID- 9201372 TI - A prospective safety-monitoring study of immunotherapy in mite-allergy patients with mass-units-standardized extract. AB - To determine the tolerance of an allergen extract standardized in mass units, we closely monitored the side-effects, during the buildup phase of immunotherapy treatment, in 88 patients with well-documented respiratory allergy to house-dust mite (Dermatophagoides pteronyssinus). Thirty-four patients (38.6%) suffered from moderate perennial rhinitis, and 54 had mild to moderate bronchial asthma (61.4%). For the desensitizing treatment, we used a depot extract adsorbed in aluminum hydroxide of D. pteronyssinus (Pangramin Depot UM), biologically standardized and having the major allergens Der p 1 and Der p 2 quantified in micrograms. A total of 1244 doses were administered. All patients except one (98.9%) reached the expected maximum dose of 4 micrograms Der p 1. Only five patients suffered mild adverse reactions (5.7%). All adverse reactions except one appeared to be related to the vial of maximum concentration: vial III (4 micrograms Der p 1). Considering the number of patients who had adverse reactions and the frequency of adverse reactions per dose, we found no significant differences between rhinitis and asthma sufferers. We think that immunotherapy in doses of 4 micrograms Der p 1 and 2 micrograms Der p 2 is well tolerated and should not be avoided in mildly to moderately asthmatic patients when treating house-dust mite allergy. PMID- 9201374 TI - Leukotriene E4 plasma levels in adult asthmatic patients with variable disease severity. AB - Cysteinyl leukotrienes (C-LTs) are local inflammatory mediators involved in bronchial asthma. Seventeen asthmatic patients (FEV1 ranging from 41 to 99.8% of predicted values) and 11 healthy subjects were studied. The clinical severity of asthma was assessed by the Aas score. Plasma C-LTs were measured by enzyme immunoassay (EIA) after sample purification by solid-phase extraction (SPE), to investigate whether differences may exist between asthmatic and control subjects and whether leukotriene E4 (LTE4) levels were related to the severity of disease. LT measurements showed that 87.6 +/- 1.2% was recovered as LTE4 and 9.4 +/- 1.3% as LTC4. In asthmatic subjects, LTE4 plasma levels were found to be significantly higher than those in the control group (1.073 +/- 0.133 and 0.53 +/- 0.19 ng/ml of plasma, respectively; P < 0.002). Moreover, there was a significant correlation between LTE4 plasma levels and the Aas clinical score (P < 0.005). These data suggest that plasma LTE4 levels might be used to assess the severity of asthma. PMID- 9201373 TI - Urinary eosinophil protein X in relation to disease activity in childhood asthma. AB - The clinical use of urinary eosinophil protein X (U-EPX) measurements in monitoring inflammation in childhood asthma was investigated. U-EPX and pulmonary function were assessed in 80 children with bronchial asthma and 24 healthy, age matched controls. In addition, 14 patients with asthma were re-examined after 1-2 months. U-EPX levels were increased in children with asthma compared with controls (median 68.4 vs 35.3 micrograms/mmol creatinine; P < 0.0001). In addition, U-EPX levels were higher in symptomatic than in asymptomatic patients (median 123.5 vs 48.9 micrograms/mmol creatinine; P < 0.0001) independent of treatment modalities (i.e., inhaled steroids or disodium cromoglycate) or atopy (median 65.1 vs 86.0 micrograms/mmol creatinine). Furthermore, U-EPX levels were significantly correlated with pulmonary function. During the follow-up period, changes in U-EPX values were significantly related to changes in pulmonary function. In conclusion, our findings demonstrate that eosinophil activation can be measured in urine in childhood asthma. Concentrations of U-EPX are related to disease activity and pulmonary function, as shown in both cross-sectional and longitudinal analyses, but are independent of atopy and treatment modalities. Measurement of U-EPX may be useful in assessing the inflammatory process and therefore in the management of childhood asthma. PMID- 9201375 TI - Ciprofloxacin-induced cutaneous vasculitis. PMID- 9201376 TI - Occupational asthma in poultry-slaughterhouse workers. PMID- 9201377 TI - Nimesulide reduces skin-test reactivity. PMID- 9201378 TI - IgE-mediated hypersensitivity to custard-apple. PMID- 9201379 TI - Immediate hypersensitivity reaction to date. PMID- 9201380 TI - Ciprofloxacin-induced vasculitis. PMID- 9201382 TI - Latency estimation of auditory brainstem response by neural networks. AB - In the clinical application of auditory brainstem responses (ABRs), the latencies of five to seven main peaks are extremely important parameters for diagnosis. In practice, the latencies have mainly been done by manual measurement so far. In recent years, some new techniques have been developed involving automatic computer recognition. Computer recognition is difficult, however, since some peaks are complicated and vary a lot individually. In this paper, we introduce an artificial neural network method for ABR research. The detection of ABR is performed by using artificial neural networks. A proper bandpass filter is designed for peak extraction. Moreover, a new approach to estimate the latencies of the peaks by artificial neural networks is presented. The neural networks are studied in relation to the selection of model, number of layers and number of neurons in each hidden layer. Experimental results are described showing that artificial neural networks are a promising method in the study of ABR. PMID- 9201381 TI - Artificial neural network analysis of noisy visual field data in glaucoma. AB - This paper reports on the application of an artificial neural network to the clinical analysis of ophthalmological data. In particular a 2-dimensional Kohonen self-organising feature map (SOM) is used to analyse visual field data from glaucoma patients. Importantly, the paper addresses the problem of how the SOM can be utilised to accommodate the noise within the data. This is a particularly important problem within longitudinal assessment, where detecting significant change is the crux of the problem in clinical diagnosis. Data from 737 glaucomatous visual field records (Humphrey Visual Field Analyzer, program 24-2) are used to train a SOM with 25 nodes organised on a square grid. The SOM clusters the data organising the output map such that fields with early and advanced loss are at extreme positions, with a continuum of change in place and extent of loss represented by the intervening nodes. For each SOM node 100 variants, generated by a computer simulation modelling the variability that might be expected in a glaucomatous eye, are also classified by the network to establish the extent of noise upon classification. Field change is then measured with respect to classification of a subsequent field, outside the area defined by the original field and its variants. The significant contribution of this paper is that the spatial analysis of the field data, which is provided by the SOM, has been augmented with noise analysis enhancing the visual representation of longitudinal data and enabling quantification of significant class change. PMID- 9201383 TI - The development and implementation of an expert system for the analysis of umbilical cord blood. AB - An assessment of neonatal outcome may be obtained from analysis of blood in the umbilical cord of the infant immediately after delivery. This can provide information on the health of the newborn infant, guide requirements for neonatal care, and is recommended practice of the Royal College of Obstetricians and Gynaecologists. However, there are problems with the technique. Samples frequently contain errors in one or more of the important parameters, preventing accurate interpretation and many clinical staff lack the expert knowledge required to interpret error-free results. In this paper the development and implementation of an expert system to overcome these difficulties is described. The expert system validates results, provides a textual interpretation and archives all results to database for audit, research and medico-legal purposes. The system has now been in routine clinical use for over 3 years in Plymouth, and has also been installed in several other hospitals in the UK. Results are presented in which the types and frequency of errors are established and the user acceptance of the system is determined. PMID- 9201384 TI - Applications of abduction: hypothesis testing of neuroendocrinological qualitative compartmental models. AB - It is difficult to assess hypothetical models in poorly measured domains such as neuroendocrinology. Without a large library of observations to constrain inference, the execution of such incomplete models implies making assumptions. Mutually exclusive assumptions must be kept in separate worlds. We define a general abductive multiple-worlds engine that assesses such models by (i) generating the worlds and (ii) tests if these worlds contain known behaviour. World generation is constrained via the use of relevant envisionment. We describe QCM, a modeling language for compartmental models that can be processed by this inference engine. This tool has been used to find faults in theories published in international refereed journals; i.e. QCM can detect faults which are invisible to other methods. The generality and computational limits of this approach are discussed. In short, this approach is applicable to any representation that can be compiled into an and-or graph, provided the graphs are not too big or too intricate (fanout < 7). PMID- 9201385 TI - BANTER: a Bayesian network tutoring shell. AB - We present an educational tool for bringing the information contained in a Bayesian network to the end user in an easily intelligible form. The BANTER shell is designed to tutor users in evaluation of hypotheses and selection of optimal diagnostic procedures. BANTER can be used with any Bayesian network containing nodes that can be classified into hypotheses, observations, and diagnostic procedures. The system enables one to present various types of queries to the network, to test one's ability to select optimal diagnostic procedures, and the request explanations. We describe the system's capabilities by illustrating how it functions with two structurally different network models of real-world medical problems. PMID- 9201386 TI - Efficacy of aluminum hydroxide-adjuvanted Escherichia coli bacterin in turkey poults. AB - Aluminium hydroxide-adjuvanted Escherichia coli bacterins were evaluated for efficacy in protecting turkeys against homologous challenge. In each of six trials involving four different E. coli serotypes, poults in one group received a single subcutaneous injection at 1 day of age, poults in a second group were vaccinated twice at 1 and 14 days of age, and those in a third nonvaccinated group served as controls. Vaccinated and control turkeys were challenged at 4 wk of age and survivors were necropsied 1 wk later. Mortality and gross lesion scores of both vaccinated groups were compared with those of the nonvaccinated group. Poults vaccinated twice had significantly lower mortality and less severe gross lesions than poults receiving no vaccine prior to challenge. Immunization with one or two injections of aluminum hydroxide-adjuvanted E. coli bacterin did not impair weight gain of poults. PMID- 9201387 TI - Experimental reproduction of a spiking mortality syndrome of turkeys. AB - Two-day-old turkey poults were inoculated with either a chicken embryo homogenate used previously to produce spiking mortality syndrome in chickens (the "Oakwood Agent") or an intestine-pancreas homogenate collected from field turkeys with the syndrome known as spiking mortality of turkeys. Twelve days postinoculation, the mean plasma insulinlike growth factor-1 (IGF-1) level and mean body weights were significantly depressed, and the mean plasma growth hormone level was significantly elevated, in the poults receiving the turkey-derived homogenate (P < or = 0.0003), as was previously reported in chickens with spiking mortality syndrome. The depression in plasma IGF-1 levels may explain the runting seen in poults that survive spiking mortality of turkeys in the field. Following a 4-hr fast and a brief cool water spraying, poults exhibited clinical signs indistinguishable from those of chicks with spiking mortality syndrome. However, plasma glucose levels in the affected poults were within the normal range, unlike chickens with spiking mortality syndrome. Immunohistochemistry on formalin-fixed intestines, ceca, and bursae produced positive staining using an arenavirus antibody in epithelial cells of poults inoculated with the turkey homogenate and those inoculated with the Oakwood Agent. Tissues of uninoculated controls were negative. Poults inoculated with the Oakwood Agent did not show noticeable disease. PMID- 9201388 TI - Isolation and identification of infectious bronchitis virus from chickens in Sichuan, China. AB - A nonhemagglutinating virus was isolated from kidneys and lungs of chickens suspected of having infectious bronchitis infection. Specific-pathogen-free embryonated chicken eggs were used as the cultural system. With the use of the ciliary activity of chicken embryo tracheal organ cultures as indicator system, the physicochemical properties of one of the isolated strains (SAIB3) were shown to be similar to infectious bronchitis virus (IBV) strain M41 (standard strain); whereas electron microscopy of the isolate showed coronavirus particles. Virus neutralization tests were performed in tracheal organ cultures to compare the serotypes of five IBV isolates and six known IBV strains on the basis of reciprocal neutralization titers and euclidean distance. The cross-neutralization pattern indicated that one isolate was of the T-strain IBV serotype, another of the M41 IBV serotype, while others had partial serotype relationship to M41 and T strains of IBV. PMID- 9201390 TI - Epitope diversity of F strain Mycoplasma gallisepticum detected by flow cytometry. AB - A culture of F strain Mycoplasma gallisepticum (F-MG) that exhibited an epitope identified by monoclonal antibody (MAb) 6F10 was used to inoculate leghorn hens in two different trials. In Trial 1, mature hens chronically infected with F-MG were swabbed at intervals from 230 to 345 days postinoculation (PI). The F-MG isolates were tested with an agar plate fluorescent antibody (APFA) method that used a polyclonal antibody and with a flow cytometry (PC) technique that used MAb 6F10. Primary cultures of swabs taken at 258, 272, 293, 318, and 345 days PI were all identified as positive by APFA, whereas FC identified 23%-41% as positive. Subsequently, MAb 6B11 was found, which reacted positively with isolates negative to MAb 6F10. Both 6F10 and 6B11 were used in the second trial, which was designed to identify F-MG isolates that were negative to 6F10. In Trial 2, naive birds were inoculated with F-MG when they were 9 wk old and were sampled at six intervals from 13 to 154 days PI. The APFA method was used to identify primary isolation (P0) cultures, and FC was performed on P0 cultures and the same cultures after they had been passed three times (third serial passage [P3] cultures). The APFA test identified 100% of the P0 cultures as F-MG. The FC results on P0 cultures showed 34.5% as 6F10 positive and 85.1% as 6B11 positive. Results for FC on P3 cultures showed 92.3% 6F10 positive and 96.3% 6B11 positive. These results suggest that the microenvironment of the colonization site in the hen induced an epitope diversity in F-MG, as evidenced by the loss in the expression of MAb 6F10-defined epitope. Isolation of the organism from hens and propagation for several in vitro passages resulted in the re-expression of the epitope defined by MAb 6F10. PMID- 9201389 TI - Recombinant env-gp85 of HPRS-103 (subgroup J) avian leukosis virus: antigenic characteristics and usefulness as a diagnostic reagent. AB - We describe the construction of a recombinant baculovirus containing the cloned DNA encoding the gp85 envelope glycoprotein of HPRS-103 (subgroup J) avian leukosis virus fused to the carboxy-terminus of the affinity tag glutathione-S transferase. The fusion protein was efficiently secreted into the supernatant medium of the infected insect cell culture and could be purified in a single step using immobilized glutathione. An enzyme-linked immunosorbent assay using the recombinant protein was found to be specific and sensitive for detection of HPRS 103 virus-specific antibodies in the sera of infected birds. PMID- 9201391 TI - Salmonella enteritidis contamination of eggs from hens inoculated by vaginal, cloacal, and intravenous routes. AB - Laying hens were inoculated intravaginally (IVg) once (IVg-single) or three times (IVg-triple), intracloacally (IC), or intravenously (IV) with Salmonella enteritidis (SE) phage type 4. Eggs tested were significantly (P < 0.05) fewer positive in group IC than in other groups. SE was recovered from egg contents in the groups IVg-single (9.6%), IVg-triple (4.2%), and IV (11.5%). IVg and IC inoculation resulted in colonization of the cloaca and lower portions of the oviduct but not the portion above the isthmus, whereas IV inoculation resulted in colonization of the entire oviduct. Only IV inoculation resulted in colonization of the ovary. In group IV, SE was recovered from three of six eggs found in the oviduct at necropsy, but in other groups, SE was not recovered from 53 eggs in the oviduct. The results suggested that the SE infection of vagina resulted in a frequent incidence of contaminated eggs and that SE adhered to the eggs from the contaminated vagina might pass through shells and shell membranes. PMID- 9201392 TI - The individual and combined effects of the Fusarium mycotoxins moniliformin and fumonisin B1 in turkeys. AB - Fumonisin B1 (FB1) and moniliformin (M) were supplied by Fusarium moniliforme M 1325 and Fusarium fujikuroi M-1214 culture material, respectively. Turkeys were fed a control ration, or rations containing 200 mg FB1/kg, 100 mg M/kg, or a combination of both 200 mg FB1/kg and 100 mg M/kg feed from 1 to 21 days of age. These rations contained 0, 3.8, 1.0, and 4.8% culture material, respectively. In comparison to controls, turkeys fed FB1 had increased relative liver weights. Both aspartate aminotransferase and lactate dehydrogenase were increased in poults fed FB1. Turkeys fed M had decreased feed intake and body weight gains and increased relative heart weights in comparison to controls. Poults fed FB1 had moderate diffuse hepatocellular hyperplasia and poults fed moniliformin had a loss of cardiomyocyte cross striations. Turkeys fed the ration containing both M and FB1 had all the above changes; however, no additive or synergistic effects were evident for any single parameter measured. No treatment-related morbidity or mortality was observed in the study. PMID- 9201393 TI - Proliferation and apoptosis in infection with infectious bursal disease virus: a flow cytometric study. AB - Programmed cell death, or apoptosis, is involved in the normal physiology of many immunocompetent organs, including lymphocytes of the bursa of Fabricius in chickens. Involvement of apoptosis has also been described in some viral diseases such as AIDS. The purpose of this work was to study the potential role of apoptosis in the pathogenesis of Gumboro disease in the bursa of Fabricius. Our results show that 1-3 days after infection of young chickens with infectious bursal disease virus, the number of apoptotic cells increases and cellularity and proliferation decrease. Because of the dynamic nature of bursal lymphocyte populations and the involvement of apoptosis in lymphocyte cell physiology, the increased level of cells undergoing apoptosis may be due to an impairment in the withdrawal of apoptotic cells. A concomitant increase in macrophages in infected bursae and a dramatic decrease in cellularity suggest that an increase in apoptosis may be an important cause of cell depletion. PMID- 9201395 TI - Natural and experimental infection of turkeys with avian paramyxovirus-7. AB - Avian paramyxovirus-7 was isolated from a natural outbreak of respiratory tract disease in male and female commercial turkey breeder flocks. The disease spread readily between different housing complexes. The virus was isolated from affected flocks and used for experimental studies in specific-pathogen-free poults. Inoculated and contact-exposed poults developed a respiratory disease characterized by rhinitis and airsacculitis. The virus was isolated from the inoculated and contact-exposed poults, and hemagglutination inhibition antibodies were detected in exposed birds. PMID- 9201394 TI - Comparison of live avirulent PM-1 and CU fowl cholera vaccines in turkeys. AB - The live avirulent PM-1 Pasteurella multocida vaccine, grown in brain-heart infusion broth, was evaluated and compared in two experiments with the Clemson University (CU) vaccine, which had been shown to be effective in preventing fowl cholera in turkeys. Experiment 1 was performed during warm environmental temperatures and Expt. 2 during cooler environmental temperatures. The PM-1 vaccine was comparable with the CU vaccine in protecting turkeys against challenge with virulent P. multocida but was considered no less virulent than the CU because turkeys died after vaccination with both the PM-1 and the CU vaccines. A significantly (P < 0.05) higher percentage of unvaccinated turkeys challenged during the cooler environmental temperatures died than did unvaccinated turkeys challenged during the warmer temperatures. A microtiter agglutination test demonstrated a significant (P < 0.01) correlation between the level of serum anti P. multocida antibody found 1 wk after vaccination and survival after challenge with virulent P. multocida in Expt. 1 and a significant (P < 0.05) correlation between these parameters in Expt. 2. However, there was a significant (P < 0.01) negative correlation between serum anti-P. multocida antibody titer 1 wk after vaccination and body weight gained 4 wk after vaccination, but before challenge, in Expt. 1, suggesting that vaccination with the live vaccines may have had a negative effect on body weight gain. At 4 wk after challenge or 8 wk after vaccination in Expt. 2, there was also a highly significant (P < 0.001) negative correlation between these parameters in the surviving turkeys. PMID- 9201396 TI - Comparison of several enzymes between normal physeal and tibial dyschondroplastic cartilage of broiler chickens. AB - Normal physeal and dyschondroplastic cartilage of broiler chickens was examined for six enzymes by isoelectric focusing in thin-layer polyacrylamide slab gels. Acid phosphatase (ACP), esterase (EST), malate dehydrogenase (MDH), and peroxidase (PRX) were present in the normal physeal cartilage but not in the dyschondroplastic cartilage. Staining intensity of glucose-6-phosphate isomerase (GPI) and triose-phosphate isomerase (TPI) was reduced in the dyschondroplastic cartilage compared with that of the physeal cartilage. Differences in the presence of these enzymes possibly demonstrated their roles in processes of bone formation, cartilage resorption, and calcification. ACP could be involved in calcification. Lack of EST and PRX may be related to the failure of vascular invasion in dyschondroplastic cartilage of afflicted birds. A deficiency of MDH and reduced GPI and TPI in dyschondroplastic cartilage may reflect a reduction in the activity of energetic metabolism, causing the dissipation of energy and necrotic cells. PMID- 9201398 TI - Histologic study of hepatic lesions in two turkey flocks. AB - Hepatic lesions were studied in two turkey flocks by euthanatizing 50 birds a week from the ages of 1 through 15 wk. Samples of liver that contained lesions and samples of duodenum, pancreas, ileum, and cecal tonsil were examined histologically. Lymphocytic infiltrations made up 82% and 75% of the hepatic lesions, and granulomas occurred in 18% and 25% of the livers. Nematode larvae were present in 12% and 15% of the hepatic lesions. PMID- 9201397 TI - Pathogenicity and diagnosis of H5N2 Mexican avian influenza viruses in chickens. AB - Chickens were inoculated with one of five H5N2 Mexican-origin avian influenza virus (AIV) isolates to determine their pathogenicity for chickens and to determine the ability of routine virologic and serologic tests to detect infections. In laboratory infections, three AIVs, H5/94, M5/94, and J12/94, produced sporadic illness and death and were categorized as mildly pathogenic. Q1/95 produced illness and death in all inoculated chickens and was categorized as highly lethal and highly pathogenic (HP). P11/94B commonly produced clinical illness, but deaths were infrequent. During the presence of clinical signs, oropharyngeal swabs were superior for isolation of AIV, but cloacal swabs were more successful after disappearance of clinical signs. Agar gel precipitin (AGP) serologic test was superior for detecting AIV infection during the clinical phase, but AGP and hemagglutinin inhibition tests were equally effective in detecting infections after recovery from clinical illness. Passage of P11/94B parent stock and selected 14-day-embryo-passed AIVs in adult hens resulted in emergence of some HP AIV derivatives. The hemagglutinin of Q1/95 and P11/ 94B parent stock and derivative AIVs had an identical proteolytic cleavage site of.... Pro-Gln-Arg-Lys-Arg-Lys-Thr-Arg-Gly, consistent with AIVs of high pathogenicity. However, no consistent differences were identified in the sequence of the hemagglutinin gene to explain the discrepancy in lethality patterns of the P11/94B AIVs. This suggests that genes other than the hemagglutinin impact the full expression of high lethality of Mexican-origin AIV infections in chickens. PMID- 9201399 TI - The effect of the flow of air on horizontal transmission of Salmonella enteritidis in chickens. AB - Horizontal transmission of Salmonella enteritidis and the effect of airflow on spreading were examined in 80 5-wk-old chickens divided into five groups. Sixteen chickens in each group were placed in four cages in a row separated by wire. One among four chickens placed in a cage at the downwind end of the row was inoculated orally with 10(9) colony-forming units of S. enteritidis. Cecal droppings, drinking water, and feed were cultured every day. Horizontal transmission was rapid in the row with low air velocity but slow in the row with high air velocity. However, in another experiment, where the inoculated chicken was situated in a cage upstream in the airflow, horizontal transmission was equally rapid whether the airflow was rapid or slow. Contamination of feed and water never preceded the appearance of positive cecal droppings. These findings suggest that the rapidity of horizontal transmission of S. enteritidis may be affected by airflow patterns. PMID- 9201400 TI - Isolation and identification of chicken infectious anemia virus in China. AB - A chicken infectious anemia virus (CIAV) isolate was obtained from broiler flocks aged 25-40 days with anemia and poor performance and was designated SR43. The CIAV isolate was resistant to treatment with chloroform and induced thymus atrophy, bone marrow aplasia, and low hematocrit values when inoculated into 1 day-old, susceptible, specific-pathogen-free chicks. CIAV-specific antigens could be demonstrated in SR43-infected MDCC-MSB-1 cells, a cell line derived from a Marek's disease lymphoma, with the use of a monoclonal antibody specific for CIAV. CIAV DNA in infected MDCC-MSB-1 cell cultures was detected by using a polymerase chain reaction assay. These findings demonstrate that CIAV is present in China. PMID- 9201401 TI - Nucleotide sequence and phylogenetic analysis of Newcastle disease virus isolates from recent outbreaks in Taiwan. AB - Portions of the hemagglutinin neuraminidase (HN) gene of Newcastle disease virus (NDV) isolates from two recent outbreaks were sequenced to investigate epidemiology of this disease in Taiwan. These NDV isolates were all viscerotropic velogenic according to the clinical lesions produced in chickens. Sequence data were obtained from 14 NDV isolates (12 from 1995 and 2 from 1984). All isolates differed in their nucleotide sequences (from 0.3 to 15.3%), and represented potentially different strains of NDV. Phylogenetic analysis revealed that these isolates are closely related to viruses isolated from Japan and Malaysia. Some viruses isolated in 1995 appeared to evolve from viruses isolated in 1984. The results suggest that the 1995 outbreak of Newcastle disease (ND) in Taiwan may have been caused by multiple strains of velogenic NDV that have cocirculated in Taiwan for some time. Moreover, NDV isolates from racing pigeons were very similar to isolates from chickens in the same period, suggesting that both domestic and free-living birds were involved in the spread of ND in Taiwan. PMID- 9201402 TI - Characterization of a nonradioactive cloned cDNA probe for detecting avian reoviruses. AB - Avian reoviruses (ARVs) cause important losses in the poultry industry. Improved tests are needed for diagnosis of ARV infections. A cDNA library was prepared from the S1133 isolate of ARV. EcoRI-adaptored cDNA molecules were ligated into the plasmid pUC19 and used to transform Escherichia coli strain DH5 alpha MCR. One cDNA clone was selected and designated HJp1. The HJp1 was labeled by random priming with digoxigenin-dUTP. This cDNA probe hybridized with the S1 gene fragment of the ARV S1133 strain in northern blot hybridization. The probe detected ARV isolates in dot-blot hybridization assays. The probe did not cross hybridize with nucleic acids extracted from mock-infected chicken embryo fibroblast cells or unrelated avian viruses. Probe HJp1 detected as little as 0.78 ng of ARV RNA. PMID- 9201403 TI - Infectious bronchitis: effect of viral doses and routes on specific lacrimal and serum antibody responses in chickens. AB - An enzyme-linked immunosorbent assay was used to measure the effect of various infectious bronchitis virus (IBV) (strain H-120) vaccine doses and routes of immunization on specific lacrimal and serum antibody responses. The results of the first trial showed that the maximum dose, 10(6) median embryo infective doses (EID50s), delivered by the ocular route elicited both a systemic and a local antibody response in the vaccinated chickens. Lower doses of vaccinal virus, 10(4) (median dose) and 10(2) (minimum dose) EID50 delivered by the same route did not induce a detectable systemic antibody response. A significant increase of IBV-specific lacrimal IgA was elicited by both the maximum and the median vaccine doses. The low vaccine dose (10(2) EID50) did not induce a detectable increase of lacrimal IgA. In a second trial approximately the same vaccine dose was administered to different chicken groups by ocular instillation, drinking water, spray, and cloaca. The results showed that all routes of vaccination tested, including the cloacal route, resulted in an increase of specific serum antibodies. Higher IgG levels were detected throughout the experimental period after vaccination by the ocular route as compared with vaccination via the drinking water. All routes of vaccination tested resulted in an increase of specific IgA in lacrimal fluid. The vaccine application methods spray, ocular instillation, and drinking water induced similar lacrimal IgA responses. PMID- 9201404 TI - Lack of correlation between microscopic lesion scores and gross lesion scores in commercially grown broilers examined for small intestinal Eimeria spp. coccidiosis. AB - Comparisons were made between microscopic lesion scores (MLSs) and gross lesion scores (GLSs) in sections from small intestine of broilers during three routine coccidiosis screenings. The duodenal and jejunal GLS were determined and recorded by different evaluators. During each screening, 2-cm sections of duodenum and jejunum were collected, and sections of intestine were then scored using a microscopic lesion scoring system. No correlation between MLS and GLS was observed in duodenum in two out of three coccidiosis screenings, and no correlation was observed between MLS and GLS in jejunum in three out of three screenings. Our findings demonstrate that, if used alone in coccidiosis screening, GLSs can underestimate infections and may not provide a true representation of the magnitude of Eimeria maxima infection within broiler flocks. PMID- 9201405 TI - Prevalence of Salmonella in municipal sewage treatment plant effluents in southern California. AB - Effluents from 12 sewage treatment plants in southern California were examined for Salmonella using a Moore swab technique. Eight of the 12 plants were positive for Salmonella when sampled at the chlorination/dechlorination site (inside the plant). Effluents from 11 of 12 sewage treatment plants were positive for Salmonella when samples were analyzed downstream of the chlorination/dechlorination site, before effluents merge with the receiving stream (outside the plant). Two of the three control sites, an urban runoff, a raw potable water reservoir, and two other sites were also positive for Salmonella. A total of 683 Salmonella isolations were represented by 11 serogroups and 54 serotypes from 26 of 32 sampling sites. Effluents from three treatment plants and one control site (raw potable water resevior) yielded Salmonella enteritidis phage type 4, in addition to other serotypes. PMID- 9201406 TI - Effects of Newcastle disease vaccines and Newcastle disease/infectious bronchitis combination vaccines on the head-associated lymphoid tissues of the chicken. AB - Ten Newcastle disease virus (NDV) and 10 NDV and infectious bronchitis virus (IBV) combination vaccines (NDV/IBV) were evaluated for their effect on the head associated lymphoid tissue (HALT) of 2-wk-old chicks. After vaccination, the chicks were subjected to an in vivo assay that measures the ability of the gland of Harder (GH) to respond to killed Brucella abortus antigen given in the eye by titering B. abortus antibodies in the tears. Following this, several sites in the HALT and trachea were examined histologically and scored for microscopic changes. The results indicated that three of the NDV/IBV combination vaccines (one BI/Mass&Conn and two LaSota/Mass&Conn) interfered with the GH response to killed B. abortus, whereas none of the NDV vaccines did Histologically, several changes were noted in the vaccinated chicks; however, no changes in the GH were observed that could explain microscopically the GH depression. With the IBV-only vaccines reported earlier (16), and the NDV-only and NDV/IBV combination vaccines reported here, a total of 36 vaccines have been evaluated using the same testing protocol. The conclusions of these combined studies suggest that several of the modified live virus vaccines containing IBV, either alone or in combination with NDV, interfere with the ability of the GH/HALT to respond to antigenic stimulation. PMID- 9201408 TI - A reproducible model for the induction of avian cellulitis in broiler chickens. AB - Avian cellulitis was reproduced in 39-day-old broilers by subcutaneous injection of Escherichia coli originally isolated from a cellulitis lesion. One hundred percent of the birds injected with the isolate on the dorsal and ventral surfaces developed characteristic fibrino-caseous plaques. A slightly lower percentage (90%) of the birds injected subcutaneously in the inguinal area developed the same lesions. Only 30% of the birds that had been inoculated by scratching the skin and swabbing the bacterial inoculum onto the wound developed the lesion. No birds inoculated by swabbing the inoculum onto a feather follicle, from which the feather had been pulled, developed cellulitis. Characteristic cellulitis plaques could be produced as early as 18 hr postinfection (PI). Lesions, consisting of a serosanguinous, yellow-pink-to-orange-tinged fluid appeared as early as 6 hr PI. The lesions progressed, changing to a more thin, yellow, purulent fluid by 12 hr PI followed by plaque formation. Although there was a trend for lesion size to diminish with time, the majority of the challenged birds, examined as late as 3 wk PI, still had prominent cellulitis plaques. Lesions in birds injected subcutaneously on the dorsal surface sometimes extended into other regions of the body, including the abdominal region, and thereby resembled the type of lesions that have previously been described as type I or hatchery-borne cellulitis. PMID- 9201407 TI - Retroviral insertional mutagenesis of a herpesvirus: a Marek's disease virus mutant attenuated for oncogenicity but not for immunosuppression or in vivo replication. AB - Our earlier studies have shown that retrovirus insertion into herpesvirus is an efficient process that engenders recombinant herpesviruses with altered biological properties. The RM1 clone is derived from the JM strain of Marek's disease virus (MDV) through retrovirus insertional mutagenesis and contains sequences of reticuloendotheliosis virus inserted at the junction of the internal repeat and unique short regions of the genome. In previous studies, the RM1 clone appeared attenuated for oncogenicity but caused marked atrophy of the thymic lobes. The present studies represent a detailed analysis of the biological characteristics of the RM1 clone in order to better understand mechanisms of oncogenicity and gene function of MDV. RM1 was almost fully attenuated for oncogenicity but retained other in vivo properties of virulent viruses such as thymic and bursal atrophy, early immunosuppression, early cytolytic infection followed by efficient replication, and contact spread--all normally absent in attenuated strains. This suggests that, for serotype 1 MDV, oncogenicity is not tightly linked with immunodepression or viral replication and that these properties may be controlled by different genes or mechanisms. The mutation was stable through serial passage of the virus in chickens as determined by molecular analysis. None of the mutant viruses demonstrated expansion of the 132-bp repeat region of the genome, indicating that such expansion is not required for attenuation. Chickens vaccinated with RM1 clones were protected against challenge with virulent MDV, and levels of protection exceeded those of other attenuated serotype 1 vaccine viruses. Thus, attenuation by selective mutation may be an advantageous strategy for development of serotype 1 Marek's disease vaccines. PMID- 9201409 TI - Evaluation of ELISA titers to infectious laryngotracheitis. AB - Infectious laryngotracheitis (ILT), a highly infectious upper respiratory disease of chickens, can cause serious economic loss in areas where the poultry industry is concentrated. To determine the antibody levels associated with vaccine administration, field challenge, and protection, six groups of 20 specific pathogen-free leghorn chickens were housed in biosecured isolation units. Individual groups served as either negative controls, vaccinated (one full dose per bird of chicken embryo origin [CEO] administered by the eyedrop method) and challenged (intratracheal administration with USDA strain ILT virus at 10(4.1) 50% embryo infective dose [EID50]), or unvaccinated and challenged with USDA strain ILT virus at various dose levels (10(2.1), 10(5.1), or 10(4.1) EID50). Chickens in each group were bled weekly, and their sera were tested for antibody using a commercially available enzyme-linked immunosorbent assay test kit. The antibody response using CEO vaccine resulted in a 400-600 geometric mean titer that appeared to be protective against severe field challenge. Negative controls had no titers, whereas vaccinated and/or challenged chickens had detectable titers within 2 wk of exposure, and these titers remained high for the next 4-7 wk. Mortality in nonvaccinated controls began at 3 days post-challenge and continued for up to 10 days. PMID- 9201410 TI - The use of feed restriction for mortality control of chickens in broiler farms. AB - Feed restriction was attempted for the control of mortality at broiler farms. Respiratory signs were observed in both restricted-feed flocks and fully fed flocks, but they were less severe in the restricted-feed flocks. The death rate in the fully fed flocks began to rise at the age of 4 wk but did not do so in the restricted-feed flocks. Mortality in the restricted-feed flocks was significantly lower than in the fully fed flocks aged from 3 to 7 wk. The economic performance with restriction feeding was better than that with full feeding as a result of improvements in viability and feed conversion rates. Feed restriction appeared to be beneficial in decreasing the death losses. PMID- 9201411 TI - Heterophil response to intraperitoneal challenge with invasion-deficient salmonella enteritidis and Salmonella-immune lymphokines. AB - The present work compared the accumulation of intraperitoneal heterophils in day of-hatch chicks following treatment with Salmonella enteritidis-immune lymphokine (ILK) and challenge with various strains of Salmonella enteritidis (SE). Day-of hatch chicks received ILK by intraperitoneal injection and were challenged 1 hr later by intraperitoneal inoculation with one of the following SE strains: a wild type, SE 890034-3; a delta cya-12 delta cyp-11 avirulent vaccine strain, chi 4357; and an invasion-deficient strain, InvA::kan, chi 4420. Four hours after challenge heterophils were recovered from the peritoneal cavity by lavage. The concentration of heterophils in the recovered lavage fluid was determined. Heterophil concentrations increased in response to challenge with each SE strain but there was a lower response to the invasion-deficient strain. The difference was statistically significant. This diminished heterophil response to challenge with invasion-deficient salmonellae supports existing evidence that the initial defensive reaction occurs at the earliest stages of the Salmonella-host interaction. PMID- 9201412 TI - Experimental reproduction of hypoglycemia-spiking mortality syndrome in broiler chickens with the use of homogenized brains containing arenaviruslike particles. AB - Severe hypoglycemia-spiking mortality syndrome was experimentally reproduced in broiler chicks. Inoculum was homogenized brains from 28-day-old commercial broiler chicks with central nervous system signs (50% [v/v] in phosphate-buffered saline with 2% fetal calf serum). Oral inoculations of 1.2 ml of the homogenate were given at 1 day of age to broiler chicks (n = 15). Fourteen days later, chicks were fasted and stressed with a 2-sec cool water spray. Six chicks (40%) developed clinical signs of spiking mortality syndrome and were severely hypoglycemic. Uninoculated control chicks (n = 15) from the same hatch, also fasted and stressed simultaneously, were unaffected. Examination of a banded fraction produced from the inoculum with the use of transmission electron microscopy with negative staining revealed viruslike particles indistinguishable from arenavirus particles stained and examined simultaneously. Avian encephalomyelitis virus was isolated by one of three laboratories attempting virus isolation with the use of embryonating chicken eggs. PMID- 9201413 TI - In situ detection of reovirus in formalin-fixed, paraffin-embedded chicken tissues using a digoxigenin-labeled cDNA probe. AB - An in situ hybridization (ISH) technique using a digoxigenin (DIG)-labeled cDNA probe detected avian reovirus (ARV) RNA in formalin-fixed, paraffin-embedded chicken tissues. Tissues were collected 3 and 10 days following inoculation with the R-2 or the S1133 strain of ARV. The cDNA clone HJp1, located on the S1 gene segment of the ARV S1133 strain, was used to prepare a nonradioactive probe. The ISH assay localized ARV RNA in infected tissues including heart, liver, intestine, pancreas, and tendon. No positive-stained cells occurred in sections from uninfected chickens. PMID- 9201414 TI - Efficacy of INOVOJECT egg injection system for delivering Marek's disease vaccine under hatchery conditions. AB - The ability of the INOVOJECT egg injection system to effectively deliver the appropriate titer of herpesvirus of turkeys (HVT) vaccine during normal hatchery operation was evaluated. The INOVOJECT machines configured for the Jamesway and Chickmaster commercial hatchers both maintained the integrity of the HVT vaccine without an appreciable loss in titer from the diluted vaccine to the egg. PMID- 9201415 TI - Application of normal avian gut flora by prolonged aerosolization onto turkey hatching eggs naturally exposed to Salmonella. AB - A commercial preparation of normal avian gut flora (NAGF) was aerosolized for an extended period over turkey hatching eggs during pipping and hatching to examine any protective effects against natural exposure to salmonellae. Turkey hatching eggs, produced by salmonellae-infected breeder flocks and hatched in a commercial hatchery with a history of salmonellae contamination, were used in two trials. In Trial 1, four doses of NAGF inoculum per hatching egg were aerosolized through an automated hatcher fogging system during the final 48 hr of the pipping and hatching process. In Trial 2, two doses of NAGF inoculum were aerosolized in a like manner. In both trials, poults were exposed to Salmonella montevideo during hatching, as indicated by samples collected at the time of pull. At day 7, treated poults in both trials were culture negative for salmonellae and control poults were culture positive for salmonellae. In Trial 1, control poults were infected with Salmonella brandenburg, and in Trial 2, control poults were infected with S. montevideo. These studies justify further critical evaluation of the protective effects of prolonged aerosolization of normal avian gut flora during pipping and hatching against salmonellae colonization in turkey poults. PMID- 9201416 TI - Inactivated vaccine for protection against duck virus enteritis. AB - Immunogenicity of inactivated tissue-culture-derived duck enteritis virus (DEV) vaccines was evaluated in white Pekin and mallard ducks. DEV from a Lake Andes outbreak was propagated in chicken embryo fibroblast cells, inactivated with beta propiolactone, and emulsified with Freund's adjuvant (FA), multiple-oil emulsion (MOE), or Squalane-pluronic L121 (L121). White Pekin and mallard ducklings were vaccinated at 2 or 3 wk of age, respectively. Challenge at 2 wk postvaccination with a virulent DEV isolated from a Long Island outbreak indicated that inactivated Lake Andes (ILA) vaccine mixed with any of the above adjuvants conferred protection, even with a single-dose inoculation. Antibody responses to vaccination, as determined by indirect enzyme-linked immunosorbent assay, showed that ILA virus with FA induced an early production of antibodies similar to that induced by commercially available modified live virus (MLV) vaccine. However, the mean anti-duck virus enteritis (DVE) IgG titers determined by multiple samplings during the first 35 days postvaccination showed titers from ILA virus with FA to be at least 10 times higher than those induced by MLV vaccine. The highest antibody titers were induced by ILA mixed with FA followed by ILA mixed with the MOE and L121. The results of this study indicated that inactivated vaccine is as efficacious as modified live vaccine in enhancing protection against virulent DEV in waterfowl. PMID- 9201417 TI - Squamous cell carcinoma-like and pox lesions occurring simultaneously in chorioallantoic membranes of chicken embryos inoculated with materials from squamous cell carcinoma and pox lesions in broiler chickens. AB - The finding of closely associated squamous cell carcinoma (SCC)-like lesions and pox lesions in chorioallantoic membranes (CAMs) inoculated with skin and palate samples taken from broilers is described. The samples were obtained from two broilers coming from different flocks that were not vaccinated against fowl pox. Both birds presented skin lesions, which were diagnosed in one bird as fowl pox, and in the other as SCC. After inoculation of CAMs with fresh tissues from both birds, histologic examination revealed, in all CAMs, lesions that were characteristic of fowl pox together with lesions consistent with those seen in the skin of broilers affected with SCC. This finding was unexpected and may shed some light on the etiology of SCC. PMID- 9201418 TI - Adenovirus-induced inclusion body hepatitis in four-day-old broiler breeders. AB - Two separate parent broiler flocks originating from the same grandparent flock experienced mortalities of 23% and 40%, respectively, in chicks between 1 and 14 days of age. Chicks affected at 4 days of age had tremors, depression, and hypoglycemia. They had pale yellow, swollen, friable livers. Pancreata were discolored and hemorrhagic. Spleens were swollen and sightly darkened. Microscopic lesions consisted of multifocal areas of acute hepatic and pancreatic necrosis with numerous basophilic intranuclear inclusions with karyomegaly. Splenic sections had severe lymphoid depletion and reticular cell and macrophage hyperplasia. An adenovirus from affected livers was isolated in chicken embryo liver cells. Serologic evidence suggests that the grandparent flock began egg production seronegative to adenovirus antibodies, was exposed during production, and, subsequently, shed adenovirus vertically to its progeny. The clinical syndrome was reproduced by injecting the isolated adenovirus into 1-day-old antibody-negative chicks. Histologic lesions in the experimentally reproduced disease cases were identical to those in the naturally occurring cases. PMID- 9201419 TI - Multiple intussusceptions in a juvenile rhea (Rhea americana) with proventricular impaction. AB - Multiple intussusceptions of the small intestine were identified in a 4-mo-old rhea (Rhea americana) that died acutely after chronically poor growth. The chick was one of a group of 12 chicks that exhibited musculoskeletal deformities, poor growth, and subsequent death. Gross necropsy findings of this chick revealed proventricular impaction by sticks and stones accompanied by multiple intussusceptions of the small intestine. This finding identifies a disease process that may afflict ratites when raised for production and that may complicate treatment of proventricular impaction. PMID- 9201420 TI - Debilitating cutaneous poxvirus infection in a Hodgson's grandala (Grandala coelicolor). AB - A case of cutaneous avian pox infection in a Hodgson's grandala (Grandala coelicolor) is described. The bird was emaciated and had nodules on the eyelids, bill, neck, legs, and toes. Eosinophilic intracytoplasmic inclusion bodies were visualized by light microscopy in epithelial cells of the cutaneous nodules. Electron microscopy revealed numerous pox virions in the inclusion bodies. This is the first report of cutaneous poxvirus infection in a Hodgson's grandala. PMID- 9201421 TI - Clinical illness in a wild turkey with Laminosioptes cysticola infestation of the viscera and peripheral nerves. AB - Laminosioptes cysticola, the fowl cyst mite, was found in peripheral nerves and thoracic and abdominal viscera of an emaciated eastern wild turkey (Meleagris gallopavo) exhibiting severe torticollis, circling, loss of balance, and wing droop. Mites, sometimes accompanied by granulomatous inflammation, were abundant in brachial plexus and sciatic nerves. Mild lymphoplasmacytic perivascular cuffing was present in the cerebellum, but no direct evidence of mites or other infectious agents was found in the central nervous system. This is the first report of L. cysticola infestation in a wild turkey and of the invasion of nervous tissue by this mite. PMID- 9201422 TI - Status of Salmonella gallinarum-pullorum infections in poultry in Zambia. AB - Ten outbreaks of Salmonella gallinarum-pullorum infections on poultry farms in Zambia were investigated. Three cases were seen in day-old broiler chickens and were diagnosed by culture as S. gallinarum-pullorum and characterized as pullorum disease because the mortality was only in the first few weeks. Another case was diagnosed by culture from broiler parent stock. Day-old chicks from two of the three cases were supplied by a hatchery. Five cases in 5-to-18-month-old layer chickens were diagnosed by culture as S. gallinarum-pullorum and characterized as fowl typhoid because of the clinical disease appearing after 5 months of age and the typical lesions of fowl typhoid. The last case was in 5-month-old village bred fowls and was diagnosed by culture and clinical manifestation as fowl typhoid. Outbreaks of S. gallinarum-pullorum are still manifest in Zambia. Clinically, both pullorum disease and fowl typhoid were observed, and it was indicated that hatchery infection plays an important role in the transmission of S. gallinarum-pullorum. PMID- 9201423 TI - Fatal avian polyomavirus infection during quarantine in adult wild-caught red faced lovebirds (Agapornis pullaria). AB - An outbreak of avian polyomavirus infection is reported in a group of six wild caught red-faced lovebirds (Agapornis pullaria), all of which died during quarantine. The birds had not shown any previous symptoms. Histologic examination of the lungs, kidneys, livers, and spleens revealed the presence of basophilic intranuclear inclusions. Avian polyomavirus was isolated from the liver and the spleen. Neutralizing antibodies were detected in the sera from other lovebirds (Agapornis personata, Agapornis taranta) that had contact with the A. pullaria. A serologic comparison showed a close relationship with budgerigar polyomavirus. PMID- 9201424 TI - Lymphosarcoma with plasmacytoid differentiation in a scarlet macaw (Ara macao). AB - A lymphosarcoma in a scarlet macaw (Ara macao) affecting periocular structures is described. Microscopically and ultrastructurally, many of the lymphoid cells had plasmacytoid features. Polymerase chain reaction amplification failed to detect exogenous avian retrovirus RAV-1 in the neoplastic mass. PMID- 9201425 TI - Chlamydiosis in captive white-winged doves (Zenaida asiatica). AB - Chlamydia psittaci was isolated from the spleen of a moribund white-winged dove (Zenaida asiatica). The isolate was serotyped as the serovar B that is commonly isolated from pigeons. A fourfold increase in the titer of antichlamydial IgM activity occurred in that bird in paired serum samples tested by chlamydial elementary body agglutination (EBA) and a greater than or equal to fourfold decrease of IgG occurred by direct complement fixation (DCF). The increases or decreases of EBA and DCF titers in other clinically ill birds that were treated with tetracycline varied, as normally occurs in cases of avian chlamydiosis. Titers in clinically normal birds were consistent with past infections. These birds were from a captive group of about 200 birds to be used for breeding and reproduction research. A small sample of recently caught wild birds was serologically negative for chlamydial antibody activity. PMID- 9201426 TI - Smoking cessation. 1: An overview of research. AB - Although tobacco smoking has long been recognized as having negative health consequences, more than one quarter of the US adult population smokes. This article presents (a) national trends in the prevalence of tobacco smoking, (b) health consequences associated with tobacco smoking and tobacco's mode of action (how tobacco/nicotine cause the problems), and (c) a brief overview of the smoking cessation treatment literature and several recommendations based on the review of research. PMID- 9201427 TI - Smoking cessation. 2: Components of effective intervention. AB - Smoking cessation treatment is an essential component of comprehensive healthcare, but many healthcare providers lack formal training and are hesitant to provide such intervention. The recently published US Agency for Health Care Policy and Research (AHCPR) Smoking Cessation Clinical Practice Guideline provided empirically based recommendations to address these issues. The most effective components of smoking cessation include the use of nicotine replacement therapy, provider support and encouragement, and training in such skills as problem solving and coping. Methods of using these recommendations are illustrated, and sample scripts are offered to serve as references for providers from various disciplines who conduct smoking cessation interventions. PMID- 9201428 TI - Smoking cessation. 3: Needed healthcare policy changes. AB - Smoking is the primary preventable cause of mortality and morbidity in our society, killing more than 430,000 people each year--more than 1,000 a day. Despite this deadly record, the treatment of nicotine dependence has not been integrated into routine medical care. Although professionals from many healthcare fields can be effective providers of smoking cessation treatment, relatively few actually advise patients to quit smoking; and even fewer assist their patients in quitting. Systematic changes in healthcare policies are needed to rectify these problems and improve the provision of smoking cessation services. In this article, the issues of who should be providing cessation treatment, why more providers do not offer this service, and what changes should be made to ensure more widespread inclusion of smoking cessation treatment in future healthcare practice are examined. PMID- 9201429 TI - Patient-provider agreement on guidelines for preparation for breast cancer treatment. AB - Guidelines for preparing cancer patients for threatening medical procedures were developed and refined and their perceived relevance and importance rated by three concerned groups--84 breast cancer patients, 64 doctors, and 140 nurses and nurse oncologists. All three groups indicated strong support for the guidelines. Patients and nurses rated more of the guidelines as essential aspects of good quality care than did doctors. Items in which a significant discrepancy existed included the importance of (a) consistent information, (b) involvement of others in preparation, and (c) assistance to the patient in coping with treatment for breast cancer. Doctors, compared with patients and nurses, underrated the importance of some aspects of preparation. These issues should be given more prominence in undergraduate and specialist medical training, as well as in continuing medical education. PMID- 9201430 TI - Serum bone alkaline phosphatase is superior to plasma levels of bone matrix proteins for assessment of bone metabolism in patients receiving renal transplants. AB - The plasma concentrations of two bone matrix proteins (osteocalcin, osteonectin) were monitored in 56 samples from 14 patients receiving renal transplants and the values compared with serum bone alkaline phosphatase mass concentrations and osteotropic hormone levels (parathyroid hormone, calcitriol). There were no significant changes in the concentrations of plasma osteonectin at any time after transplantation, as compared with the values before transplantation (P > 0.1). None of the plasma samples showed osteonectin levels above the reference interval. There was a weak but significant relationship between platelet counts and plasma osteonectin levels (r = +0.322; P < 0.05). Osteocalcin showed a marked decrease of the values 1 week following transplantation as compared with the values before transplantation without further change of the values 1 and 3 months after transplantation (P > 0.5) whereas 3 months after transplantation bone alkaline phosphatase levels were higher than before transplantation (P < 0.05). Multiple regression analysis (performed with data from 42 samples obtained after transplantation) revealed serum creatinine as an independent predictor of plasma osteocalcin whereas serum calcitriol was an independent predictor of serum bone alkaline phosphatase (P < 0.05). No correlation was observed between serum calcitriol/plasma parathyroid hormone on the one hand and plasma osteocalcin on the other (P > 0.05). After transplantation there was a lack of correlation between serum bone alkaline phosphatase mass concentrations and plasma osteocalcin values (P > 0.05). In conclusion, serum bone alkaline phosphatase should be preferred to bone matrix proteins for the assessment of bone metabolism in patients receiving renal transplants: (a) bone alkaline phosphatase-but not osteocalcin-is significantly correlated with calcitriol and adequately reflects increased bone formation after renal transplantation; (b) interpretation of osteocalcin values is severely hampered by their strong correlation with serum creatinine concentrations; (c) plasma osteonectin determinations are not useful for monitoring bone formation. PMID- 9201431 TI - Extracellular glutathione peroxidase and ascorbic acid in aqueous humor and serum of patients operated on for cataract. AB - Patients operated on for cataract (32 men/75 women, aged 50-93 years) were studied with respect to antioxidative agents in aqueous humor and serum. Extracellular glutathione peroxidase (eGSHPx) was demonstrated in aqueous humor for the first time by a radioimmunoassay, the concentration of eGSHPx being 0.66(0.18) mg/l (mean(S.D.)). The concentration of eGSHPx in serum was 3.81(0.84) mg/l, and its level in aqueous humor was 18(7)% of that level. Serum selenium had positive correlations with both serum eGSHPx (r = 0.34, P < 0.001) and aqueous humor eGSHPx (r = 0.25, P = 0.011). However, there was no relation between the concentrations of eGSHPx in aqueous humor and in serum, suggesting that the maintenance of eGSHPx levels in the two fluids is controlled by different mechanisms beside selenium status. There was an inverse correlation between age and serum eGSHPx but not with aqueous humor eGSHPx. The concentration of ascorbic acid in aqueous humor was 2.04(0.58) mmol/l, and it was closely correlated to the level of ascorbic acid in serum (0.052(0.032) mmol/l), r = 0.58 (P < 0.001). The ratio between the level of ascorbic acid in aqueous humor and that in serum was 39(17). There was no significant difference among patients with nuclear (n = 39), cortical (n = 20), posterior-subcapsular (n = 23) or mixed (n = 23) lens opacity with respect to levels of eGSHPx and ascorbic acid in serum and aqueous humor. Since serum ascorbic acid is related to ascorbic acid intake, its association to aqueous humor ascorbic acid indicates that dietary habits are important for maintaining that level which could play an important role in protecting ocular tissue against oxidative damage. The role of eGSHPx secreted into aqueous humor in the oxidant defence system needs further study. PMID- 9201432 TI - An improved HPLC-enzyme-reactor method for the determination of oxalic acid in complex matrices. AB - In this paper we present an improved method for the selective and sensitive determination of oxalate in different matrices such as urine, plasma, and food. The method uses ion chromatography for the separation of anions. To overcome problems with interfering matrix-anions, colourings, and macromolecules, we used an inline enzyme-reactor (ER) containing immobilised oxalate oxidase, which converts oxalate to hydrogen peroxide. Hydrogen peroxide was analysed with high sensitivity by amperometric detection. The determination limit for the HPLC-ER method was 1.5 mumol/1, the mean recovery in urine was 102%. The evaluation in a urinary matrix achieved C.V. values from 2.2% to 6.7% for the within-run precision and C.V. values from 3.7% to 8.6% for the between-batch precision. The results of the new method were statistically equivalent to those obtained by enzymatic kits. We present first results of the HPLC-ER method, when applied to body fluids and food analysis. PMID- 9201433 TI - The fat emulsion agglutination test: a reliable and cost effective alternative to the latex agglutination test for rapid bedside CRP measurement. AB - We compared two tests for bedside C reactive protein (CRP) measurement: the latex agglutination test (LAT) and the fat agglutination test (FAT). FAT is based on the property of CRP to agglutinate fat emulsions in the presence of CaCl2. The sensitivity, specificity and accuracy of FAT and LAT to detect a CRP > 10 mg/l, determined with radial immunodiffusion (n = 500 pediatric patients, CRP range 0- > 80 mg/l), were 91%, 82% and 90% respectively for FAT and 82%, 95% and 85% for LAT. FAT reagent could be stabilized with NaN3 (0.02%) for at least one year, when stored at 4 degrees C (n = 49). NaN3 (0.02%) had no effect on agglutination of FAT (n = 40). In conclusion, in pediatric patients, FAT is a reliable and cost effective alternative to LAT, if serum samples are used. PMID- 9201434 TI - Hyperglycemia-induced hyperinsulinemia acutely lowers plasma apolipoprotein B but not lipoprotein (a) in man. AB - Acute hyperinsulinemia lowers plasma apolipoprotein B (apo B) and triglycerides by suppressing hepatic lipoprotein secretion and probably by enhancing catabolism of triglyceride-rich lipoproteins, but the effect of acute hyperinsulinemia on the plasma lipoprotein(a) (Lp(a)) level is unclear. We measured plasma triglycerides, cholesterol, apo B and Lp(a) in response to 3 h hyperglycemia induced hyperinsulinemia (blood glucose clamped at 10 mmol/l) in 16 subjects (eight women and eight men). In a control experiment saline was infused in another group of seven men. After 3 h of hyperinsulinemia plasma triglycerides decreased by 29 +/- 14% (mean +/- S.D., P < 0.001) and this fall differed from the unchanged triglyceride level during saline infusion (P < 0.001). Plasma cholesterol fell by 8 +/- 5% (P < 0.001), which was different from the unchanged cholesterol during saline infusion (P < 0.02). Plasma apo B decreased by 9 +/- 8% (P < 0.001), which was again different from the minor fall in apo B (3 +/- 2%) during saline infusion (P < 0.02). However, plasma Lp(a) remained unchanged during hyperinsulinemia (change 8 +/- 15%, n.s.), as well as during saline infusion (change 5 +/- 15%, n.s.). The % change in apo B exceeded the % change in Lp(a) during hyperinsulinemia (P < 0.01). Baseline Lp(a) was inversely correlated with first phase insulin secretion (P < 0.05), but its level during the clamp was not related to insulin sensitivity. This study demonstrates that acute hyperglycemia-induced hyperinsulinemia has a different effect on plasma apo B and Lp(a) in healthy subjects. The present data support the notion that Lp(a) is metabolized differently from triglyceride-rich lipoproteins. PMID- 9201436 TI - Paradoxical behaviour of lyophilised commercial control materials for CK and CK MB assays after reconstitution at either 4 degrees C or 24 degrees C. PMID- 9201435 TI - Blood lipids of patients with chronic hepatitis: differences related to viral etiology. AB - In order to investigate whether a difference might exist in blood cholesterol and its subtractions between patients with chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infection, serum cholesterol, HDL-cholesterol, triglycerides and common liver function tests were measured in 138 patients (92 male, 46 female) with biopsy-proven chronic viral hepatitis without cirrhosis. Twenty-four had hepatitis B and 114 hepatitis C. Mean serum cholesterol was lower in HCV-infected in comparison to HBV-infected patients (175 +/- 36 mg/dl vs. 189 +/- 28 mg/dl, p < 0.05). On multivariate analysis, etiology of hepatitis appeared to be associated with the value of serum cholesterol, independently of age, sex and liver synthetic function (improvement of chi-square 4.40, p < 0.05). In patients with HBV infection, circulating tumor necrosis factor-alpha demonstrated a correlation with serum triglycerides (p = 0.618) and an inverse correlation with serum HDL-cholesterol (p = -0.456); in the group of patients with HCV infection, interleukin-6 correlated with triglycerides (p = 0.370) and HDL cholesterol (p = -0.355). Thus, differences in the mechanisms of liver damage and of viral clearance in hepatitis C in comparison to hepatitis B, reflected in these patients by the levels of circulating cytokines, may be mirrored by differences in their blood lipid composition. PMID- 9201437 TI - Evaluation of a new automated latex agglutination assay for lipoprotein(a): comparison with a manual ELISA. PMID- 9201438 TI - Is echinocytic transformation of erythrocytes related to lipid peroxidation in hemodialysed uraemic patients? PMID- 9201439 TI - IgG subclass typing by immunofixation. PMID- 9201440 TI - Desferrioxamine infusion due to heparin interference with iron assay. A paediatric problem. PMID- 9201441 TI - Examining the conceptual and scientific underpinnings of research in developmental psychopathology. PMID- 9201442 TI - The Hubble hypothesis and the developmentalist's dilemma. AB - Developmental psychopathology stands poised at the close of the 20th century on the horns of a major scientific dilemma. The essence of this dilemma lies in the contrast between its heuristically rich open system concepts on the one hand, and the closed system paradigm it adopted from mainstream psychology for investigating those models on the other. Many of the research methods, assessment strategies, and data analytic models of psychology's paradigm are predicated on closed system assumptions and explanatory models. Thus, they are fundamentally inadequate for studying humans, who are unparalleled among open systems in their wide ranging capacities for equifinal and multifinal functioning. Developmental psychopathology faces two challenges in successfully negotiating the developmentalist's dilemma. The first lies in recognizing how the current paradigm encourages research practices that are antithetical to developmental principles, yet continue to flourish. I argue that the developmentalist's dilemma is sustained by long standing, mutually enabling weaknesses in the paradigm's discovery methods and scientific standards. These interdependent weaknesses function like a distorted lens on the research process by variously sustaining the illusion of theoretical progress, obscuring the need for fundamental reforms, and both constraining and misguiding reform efforts. An understanding of how these influences arise and take their toll provides a foundation and rationale for engaging the second challenge. The essence of this challenge will be finding ways to resolve the developmentalist's dilemma outside the constraints of the existing paradigm by developing indigenous research strategies, methods, and standards with fidelity to the complexity of developmental phenomena. PMID- 9201443 TI - The book of names: DSM-IV in context. AB - The authors review the constraints of current mental disorder classification systems that rely upon descriptive symptom-based approaches, and weigh the benefits and hazards of these classification and diagnostic strategies. By focusing principally on superficial descriptions of symptoms, current systems fail to address the complex nature of persons' transactions within and adaptations to difficult environments. While attempting to be atheoretical, current systems exclude types of information that may elucidate individuals' functioning across various contexts, often because it is difficult to obtain such data reliably. With current approaches, misdiagnosis is likely, particularly when diagnostic criteria are applied to persons in nonclinical settings. Alternative approaches that take fuller advantage of clinicians' expertise and other forms of clinical data are reviewed, and recommendations are made for the next generation of classification systems. Application of evolutionary theory to psychiatry and psychology, as well as development of a theory and nosology of context in terms of persons' adaptations, are needed to expand our knowledge of normal and abnormal human development and psychopathology. PMID- 9201444 TI - Psychopathology as an outcome of development. AB - When maladaptation is viewed as development rather than as disease, a transformed understanding results and a fundamentally different research agenda emerges. Within a developmental perspective, maladaptation is viewed as evolving through the successive adaptations of persons in their environments. It is not something a person "has" or an ineluctable expression of an endogenous pathogen. It is the complex result of a myriad of risk and protective factors operating over time. Key research questions within this framework center on discovery of factors that place individuals on pathways probabilistically leading to later disturbances and factors and processes which maintain individuals on, or deflect them from, such pathways once enjoined. There is an interest in recognizing patterns of maladaptation which, while not properly considered disorder themselves, commonly are precursors of disorder and also in conditions of risk that lie outside of the individual, as well as any endogenous influences. Likewise, there is a focus on factors and processes that lead individuals away from disorder that has emerged, which goes beyond interest in management of symptoms. Finally, many topics that currently are capturing attention in the field, such as "comorbidity" and "resilience," are seen in new ways from within the perspective of development. PMID- 9201445 TI - When is development disordered? Developmental psychopathology and the harmful dysfunction analysis of mental disorder. AB - One goal of developmental psychopathology is to understand the origins and course of mental disorders. I argue that pursuit of this goal requires a valid conceptual understanding of disorder and that this understanding can be provided by the "harmful dysfunction" analysis of the concept of disorder. The harmful dysfunction analysis holds that a disorder is a condition that is both harmful according to social values and caused by an internal dysfunction, that is, by a failure of an internal mechanism to perform a function for which it was naturally selected. This analysis explains why many of the distinctive features of developmental psychopathology are appropriate to the study of disorder. It is argued that the harmful dysfunction analysis is a necessary supplement to other proposed criteria for disorder, such as developmental deviation or predictive validity. PMID- 9201446 TI - A person-oriented approach in research on developmental psychopathology. AB - There is a growing acceptance of a holistic, interactionistic view in which the individual is seen as an organized whole, functioning and developing as a totality. This view emphasizes the importance of patterns of operating factors. Within this framework, a standard variable-oriented approach, focusing on the variable as the main theoretical and analytical unit, has limitations. A person oriented approach would often be preferable, where the main theoretical and analytical unit is the specific pattern of operating factors. Such an approach is presented here, focusing on individual development and psychopathology. A brief theoretical and methodological overview is given and a classification approach is emphasized. Empirical examples concerning the longitudinal study of adjustment problems illustrate a number of issues believed to be important to development and psychopathology: problem gravitation, the significance of single variables and of patterns, the developmental study of syndromes (= typical patterns), and the detection of "white spots" in development. PMID- 9201447 TI - Conceptualizing psychopathology: the importance of developmental profiles. AB - This paper examines critically four habits of social scientists: a reliance on constructs for processes that do not specify the class of agent or the context of action, skepticism toward empirical truths that are inconsistent with political or social goals, a reluctance to use profiles or characteristics that have different weights to classify persons, and an aversion to treating subjects with extreme values as representing special categories. These points are made in the context of the author's research on temperaments in children. PMID- 9201449 TI - Epigenetic approaches to developmental psychopathology. AB - We evaluate the usefulness of 11 key epigenetic concepts from the behavior genetic research paradigm for advancing the field of developmental psychopathology. Key assumptions, empirical examples, and caveats in interpreting results are presented. We emphasize the usefulness of incorporating both dimensional (e.g., temperament trait) and categorical (e.g., diagnosis) variables, environmental measures, direct behavioral assessments, and multiple, theoretically relevant occasions of study into classic twin and family studies. We highlight contemporary techniques for identifying specific chromosomal regions associated with behavioral patterns, and the importance of considering nonmendelizing genetic influences when discerning the panorama of genetic influences on behavior. PMID- 9201448 TI - Integrating nature and nurture: implications of person-environment correlations and interactions for developmental psychopathology. AB - The developmental interplay between nature and nurture is discussed, with particular reference to implications for research in developmental psychopathology. The general principles include individual differences in reactivity to the environment, two-way interplay between intraindividual biology and environmental influences, and the need to consider broader social contextual features. Individuals actively process their experiences; they also act on their environment to shape and select their experiences, and individual characteristics change over time. Key findings on genetic effects include their ubiquitous influence, the multifactorial origin of most psychopathology, the involvement of several genes in most mental disorders, some genetic effects operate through dimensional risk features rather than directly on disorder, some genetic effects are dependent on gene-environment correlations and interactions, and genetic effects increase with age. Key findings on environmental effects include their ubiquitous influence, the genetic mediation of some supposed environmental effects, the importance of passive gene-environment correlations, the paucity of evidence regarding environmental effects on lifetime liability to psychopathology, the lack of understanding of environmental effects on the organism, and the importance of nonshared environmental effects. Research strategies to investigate environmental risk mediation include the range of genetically sensitive designs, migration studies, secular trend investigations, studies of nonfamilial environments, and examination of intraindividual change in relation to measured environmental alterations. Proximal processes involved in person-environment interplay are discussed in relation to person-environment interactions and evocative and active person-environment correlations. PMID- 9201450 TI - From impasse to insight in autism research: from behavioral symptoms to biological explanations. AB - The incomplete interface between remediation-oriented research and basic science research has hampered progress toward gaining insight into the etiologies of autism, despite the availability of abundant research data. Investigators of these two research domains differ in their background training and primary goals, which necessarily affect their missions, perspectives, research questions posed, methodologies selected, and interpretation of data from the research. Miscommunication between the two types of researchers has brought about disagreement on nearly every aspect of the research process. We discuss both sides of the impasse: a traditional clinical practice perspective based on the requirement for finding immediate answers to the remediation question and the basic science perspective with the goal of delineating the sequence of biological changes from the initial cause(s) of abnormal development to behavioral outcome. Although remediation-oriented research aims at alleviation of symptoms for today's patients, we propose that a basic science perspective seeks insight into the triggering causes and pathogenesis of the disorder from which better diagnosis and remediation may be devised for patients in the future. We suggest that research in autism can progress beyond the impasse of disagreement and competition toward information integration and insight by means of dialogue, data exchange, discussion, collaboration, and cooperation. PMID- 9201451 TI - What should be the focus of emotion regulation in children? A nonlinear dynamic mathematical model of children's peer interaction in groups. AB - This paper questions the assumption that children's social and emotional competence be placed within the developing child, rather than in the interaction of the child with the range of peer social ecologies in which the children might function. This paper presents a new nonstatistical mathematical approach to modeling children's peer social interaction in small groups using nonlinear difference equations in which both an uninfluenced and an influenced regulatory set point of positive minus negative interaction can be separately estimated. Using this model and the estimation procedure, it is possible to estimate what a focal child and the group initially brings to the group interaction and also how these regulatory set points are influenced by the interaction to determine two influenced regulatory set points. Six-person mainstreamed and specialized groups were established involving three types of unacquainted preschool boys: children with and without developmental delays and a language disordered but intellectually normally functioning group, using a methodology that ensured appropriate matching of child and family characteristics. For each 2-week play group, the social interactions of each child were observed during a designated free play period. Handicapped children were observed in either a specialized or mainstreamed setting. The application made of this modeling process in this paper is generating theory to attempt to understand influence processes. Parameters are introduced that reflect uninfluenced target child and group set points, emotional inertia, and influence functions. PMID- 9201452 TI - Conceptualizing and scaling the developmental structure of behavior disorder: the Lifetime Alcohol Problems Score as an example. AB - We contrast the current, clinically based framework for behavior disorder against a life course framework, as an alternative structure upon which to map the variations in onset and stability of clinical symptomatology known to take place in adult life. This alternative developmental framework is used as a base around which to understand known variations in rates of alcohol abuse/dependence over the life course and to review existing schemes for the evaluation of developmental variation in "caseness." From this work, it was proposed that symptom structure be regarded as a mass of greater or lesser breadth, with properties of extensiveness in time and life course invasiveness, as a function of where in the life course the symptomatology first emerged, and the degree to which the mass sustained itself in developmental time. This framework guided the construction of a time-based measure of alcohol related symptomatology, called the Lifetime Alcohol Problems Score (LAPS). The LAPS discriminated among a variety of alcohol-specific and nonalcohol-specific measures of alcohol-related difficulty, including diagnosis of alcohol dependence, having been in treatment, level of other psychopathology, and measures of family disorganization. The measure has potential applicability for prospective studies, and in estimating clinical prognosis. The utility of the paradigm as a framework within which to conceptualize the emergence, ebb, and flow of other behavior disorders is also discussed. PMID- 9201453 TI - The Carhart Memorial Lecture, American Auditory Society, Salt Lake City, Utah 1996. Phoneme and word recognition for words in isolation and in sentences. AB - OBJECTIVE: To evaluate relations among scores for phonemes, words in isolation, and words in sentences for listeners with normal hearing and for listeners with sensorineural hearing loss. DESIGN: Ten-word lists of consonant-vowel-consonant monosyllables with each list utilizing the same 10 vowels and 20 consonants (Boothroyd, 1968) were devised and recorded. These words also were incorporated into contextually correct sentences and recorded by the same talker. The materials were presented in quiet to 36 listeners with normal hearing and to 876 listeners (1260 ears) with sensorineural hearing loss. Formulae derived by Boothroyd and Nittrouer (1988) to relate scores for phonemes, words, and sentences were applied to the data. RESULTS: Phoneme scoring yielded scores that were on the order of 20% higher than scores for whole words heard in isolation, and scores for words in sentences were about 20% higher than when the same words were heard singly. Relations among scores for phonemes, words in isolation, and words in sentences were very similar to those observed by Boothroyd and Nittrouer (1988). The constants derived from application of their formulae to our data were very similar to the constants Boothroyd and Nittrouer obtained for a different set of materials presented against a noise background to listeners with normal hearing. Further, the constants were similar for our group of listeners with normal hearing and our large sample of listeners with sensorineural hearing loss. CONCLUSIONS: 1) These findings support Bilger's (1984) unifying assumptions that speech recognition is a single construct; therefore, scores on all speech recognition tests must be related and scores on one speech recognition test should be predictive of scores on other tests. 2) Advantages of phoneme scoring include: A) It increases the sample size of scored items for a given list of words, thereby reducing variability in test results. B) Statistical equivalence of phoneme scores for the same 30 phonemes in each of two isophonemic word lists can be evaluated quickly and easily by applying the binomial distribution model to the scores (Thornton & Raffin, 1978). C) Phoneme scores are reasonably accurate predictors of recognition of words in the contextually correct but generally low probability sentences used in this study. PMID- 9201454 TI - Audiological correlates of speech understanding deficits in elderly listeners with mild-to-moderate hearing loss. III. Factor representation. AB - OBJECTIVE: The objective of the study was to determine the major factors that underlie auditory/audiological performance measures in an elderly population, with particular emphasis on finding those factors responsible for speech understanding under specific conditions of interference. DESIGN: Audiological status and auditory performance of a group of elderly (60- to 81-yr-old) and normal-hearing young (18- to 30-yr-old) individuals was determined through a test battery. When present, the hearing loss of elderly subjects was symmetrical in the two ears and, at most, moderate. The battery included tests of speech intelligibility on the word and sentence levels, with and without the presence of interfering speech. In addition to pure-tone and speech reception thresholds, perception of spectrally or temporally distorted speech as well as auditory resolution of frequency, time, and space were tested. Two tests received special consideration: the Speech Perception In Noise Test and the Modified Rhyme Reverberation Test. Taking the overall results as well as various subsets of the results, principal component analyses were conducted to identify major factors underlying auditory performance. RESULTS: The factors extracted by the principal component analyses present a portrayal of the auditory performance profile in which effects of interference, high-frequency hearing, and basic auditory function play a major role. Interference factors include general susceptibility to noise as well as segregation of concurrent speech sounds on the basis of temporal dissimilarities and spatial separation. Comparison of factors extracted from various subsets of tests indicate that factors underlying the decline of the "cocktail party effect" in the elderly are addressed mostly by tests specifically designed to assess speech understanding in spatially distributed babble or in a reverberant environment. CONCLUSIONS: Factor analysis of test measures obtained from a group of elderly individuals with normal hearing or mild-to-moderate hearing loss led to two main findings. First, it portrayed hearing loss as a component of different factors rather than as a factor on its own. Second, the independence of measures of speech understanding in babble or reverberation from other measures suggests that such tests should become an integral part of audiological test batteries designed to assess auditory functions in aging. PMID- 9201455 TI - Effect of set size and method on speech reception thresholds in noise. AB - OBJECTIVE: The threshold for speech is known to improve as signal (Miller, Heise, & Lichten, 1951) or response (Pollack, 1959) uncertainty is decreased. The definition of threshold as the signal level or signal to noise ratio (S/N) at which a fixed percent-correct score is obtained, therefore, becomes problematic when set size, M, is varied through the range for which threshold varies systematically (M = 2 to M = 16) because the meaning of that fixed percent changes as M is increased or decreased. The goal of the present study is to examine the effect of set size, M, on speech reception thresholds (SRTs) under two testing strategies. DESIGN: SRTs were obtained in the presence of 80 dB SPL white noise for sets containing 2, 4, 8, and 16 words using two different procedures, one in which threshold was based on a fixed percent correct (American Speech-Language-Hearing Association, 1988) and one in which threshold was defined in terms of d' = 1.00. The subjects were 12 young women with normal hearing and little or no experience with audiologic testing procedures. RESULTS: When threshold was based on a fixed percent correct, S/N at threshold was found to be dependent on set size (F = 3.333; df = 3, 33; p = 0.031). When threshold was defined in terms of d' = 1.00, S/N at threshold was found to be independent of set size. CONCLUSIONS: If a smaller set size than that recommended by the American Speech-Language-Hearing Association (1988) guidelines is to be used for obtaining SRTs in a clinical setting, thresholds should be based on a criterion free measure that is independent of the size of the set of words being tested, if possible. PMID- 9201456 TI - A tinnitus problem questionnaire in a clinic population. AB - OBJECTIVE: The purpose of this study was to investigate difficulties associated with tinnitus in a large sample of patients using an open-ended problem questionnaire. DESIGN: Four hundred seventy-three unselected patients referred to the Tinnitus Clinic of the Welsh Hearing Institute (1986 through 1991) completed the Tinnitus Problem Questionnaire (Tyler & Baker, 1988). Before clinical assessment, patients were invited to list problems that they associated with their tinnitus in order of importance in an open-ended questionnaire. Their responses were coded into 45 categories. The responses of 39 patients who only described their tinnitus were excluded, leaving unselected responses from 436 patients, 224 women and 212 men, whose average age was 57.1 yr. The questionnaire elicited an average 3.78 responses per patient (range 1 to 12). RESULTS: The 30 most common difficulties attributed to tinnitus are described. There was broad agreement between the problems reported in this clinic-based study and those reported by a self-help group sample (Tyler & Baker, 1983). CONCLUSIONS: Analysis of these responses into groups based on psychological (30.1%), hearing (23.5%), health (20.7%), sleep (14.6%), and situational (11.1%) difficulties highlights the need for recognition of the global consequences of tinnitus to many patients and for a broadly based tinnitus management model. PMID- 9201457 TI - Effect of negative middle-ear pressure on transient-evoked otoacoustic emissions. AB - OBJECTIVE: The purpose of the study was to illustrate the effect of negative middle-ear pressure (MEP) on both the stimulus and response of transient-evoked otoacoustic emissions (TEOAEs) and the effect of compensating for negative pressure in the middle ear by pneumatically introducing pressure into the ear canal. Simulation of negative MEP by introducing positive pressure into the ear canal also was examined. DESIGN: TEOAEs were measured over 6 mo in a subject who frequently had negative MEP out to -150 daPa. Compensation was done for MEPs of 105, -135, and -165 daPa. Simulation of negative pressure was done for these same pressures. The effect of a pressure differential across the eardrum on the stimulus spectrum was measured at 100, 200, and 300 daPa. All measurements were made on the same subject. RESULTS: Small amounts of negative MEP significantly affected both stimulus and response spectra. The simulated negative MEP approximated actual MEP at MEPs of -105 and -135 daPa. At -165 daPa, a divergence between the two spectra occurred below 2.0 kHz. Compensation for negative MEP by pneumatically introducing pressure into the ear canal essentially returned both spectra to that seen when the MEP was close to ambient pressure, at least for frequencies above 1.5 to 2.0 kHz. At lower frequencies, compensation resulted in increased TEOAE amplitude relative to the amplitude at ambient pressure. CONCLUSIONS: Small amounts of negative MEP may affect TEOAE spectra and potentially influence the reliability of the test. For long-term monitoring of TEOAEs, MEPs either should be near ambient pressure or should be compensated for by an equivalent pressure in the ear canal. PMID- 9201458 TI - A role for otoacoustic emissions in screening for hearing impairment and middle ear disorders in school-age children. AB - OBJECTIVE: The primary purpose of this study was to investigate the potential role of transient-evoked otoacoustic emissions (TEOAEs) for screening for hearing impairment and middle ear disorders in school-age children. Because TEOAEs are present in ears with normal cochlear and middle ear function and typically are absent or reduced in ears with cochlear and/or middle ear disorders of even mild degree, TEOAE screening could serve as a first-stage screening to separate from the general population of school-age children those at greater risk for hearing impairment and/or middle ear disorder. There were two secondary objectives. First, the relationship between TEOAE measurement variables and measures of middle ear immittance in ears declared clinically normal was investigated. Second, the performance of TEOAEs in screening was compared with the performance of the pure-tone hearing and tympanometric screening protocol commonly used in the schools. DESIGN: Sixty-six children (ages 5 to 10 yr) participated. TEOAEs, pure-tone hearing screening, acoustic immittance (single-frequency and multi frequency tympanometry), and an otoscopic exam by a pediatrician, who previously had been "validated" for identification of middle ear effusion, were done on each child under typical school hearing screening conditions. Performance of the TEOAE screening was determined based on the pediatrician's determination of middle ear status and the pure-tone hearing screening as the gold standards. RESULTS: Of the 66 subjects, 61 completed the study. Fifty-six children passed the hearing and otoscopic screenings bilaterally, and five children did not pass either or both the hearing screenings or otoscopic examination in at least one ear. A variety of TEOAE criteria were examined with respect to their ability to identify ears with either hearing impairment and/or middle ear disease. Several different otoacoustic emission criteria performed well according to our diagnostic criteria. Correlations between TEOAE variables and immittance measures of middle ear function were all low. In addition, tympenometric data were used to compare the TEOAE screening with the American Speech-Language-Hearing Association's (ASHA) recommended protocol for the same ears. The ASHA protocol, as recommended, did not do as well as the TEOAE screening. Using slightly modified criteria, the ASHA protocol did as well as TEOAEs. CONCLUSION: There were some screening criteria based on TEOAE measurement that produced good sensitivity and specificity. A TEOAE screening for hearing impairment and middle ear disease performed as well as or better than the ASHA-recommended protocol, which requires a minimum of two different tests, even when the ASHA protocol was modified to optimize performance. The results suggest that the TEOAE test has the potential to be incorporated successfully into hearing screening programs for school-age children and may have advantages over current screening protocols. Finally, no relationship between TEOAEs and middle ear function, as measured using single frequency and multifrequency tympanometry, could be determined in ears with normal hearing and normal middle ear function. PMID- 9201459 TI - Cochlear implantation of children with minimal open-set speech recognition skills. AB - OBJECTIVE: The purpose of this study was to evaluate the postoperative performance of 12 children who demonstrated some open-set speech recognition skills before receiving a Nucleus multichannel cochlear implant with a view toward expanding the selection criteria for cochlear implant candidacy to include children who derive minimal benefit from amplification. DESIGN: Pre- and postoperative performance of two groups of children were compared. Group 1 consisted of 12 children who demonstrated some open-set speech recognition skills before receiving a Nucleus multichannel cochlear implant (Borderline group). Group 2 consisted of 12 children who demonstrated no open-set speech recognition skills before implantation with a Nucleus device (Traditional group). In all children, candidacy was determined based on preimplant binaural aided performance. For most subjects, the poorer ear was selected for implantation. Mean pre- and postoperative speech recognition scores of the Borderline subjects were compared to determine the benefit provided by their cochlear implants. Secondly, matched-pair analyses were used to compare the mean speech recognition scores obtained by the Borderline and Traditional subjects. RESULTS: The scores of the Borderline group improved significantly on five of six speech recognition measures when 6 mo postoperative scores obtained with the implant were compared with preoperative test scores obtained with hearing aids. By the 12 mo postoperative interval, the scores of the Borderline group had improved significantly (p < 0.05) on all six measures. In contrast, scores obtained by the Traditional group had improved significantly on three of six measures at both the 6 and 12 mo postoperative intervals. Comparison of postoperative test scores revealed that the Borderline group scored significantly higher than the Traditional group on three of six measures at the 6 mo test interval and on six of six measures at the 12 mo test interval (p < 0.05). CONCLUSIONS: The findings of this study indicate that both groups derive significant benefit from their cochlear implants. Although the mean preoperative audiograms for the implanted ears did not differ significantly for the two groups of subjects, members of the Borderline group exhibited significantly better speech recognition skills than the Traditional group during the first year after implantation. These findings suggest that the increased auditory experience of the Borderline subjects positively influenced their performance with a cochlear implant. The authors advocate that the selection criteria used to determine pediatric cochlear implant candidacy be broadened to include consideration of children who demonstrate minimal open-set speech recognition skills. PMID- 9201460 TI - Perception of rhythmic and sequential pitch patterns by normally hearing adults and adult cochlear implant users. AB - OBJECTIVE: This study compares the musical perception of 17 adult recipients of the Nucleus cochlear implant using two different formant extraction processing strategies (F0F1F2 and MPEAK). DESIGN: Over a 12 mo period, participants were alternately switched between two strategies every 3 mo. Performance was evaluated using three measures of rhythmic and sequential pitch perception. RESULTS: Three individuals performed significantly better with the MPEAK strategy on one particular rhythm task, 11 participants performed better with the MPEAK strategy on another rhythm task, and no significant differences were found between the two strategies on a sequential pitch pattern task. CONCLUSIONS: Neither strategy seems clearly superior for perception of either sequential pitch or rhythmic patterns. PMID- 9201461 TI - Comparison between esophageal Wallstent and Ultraflex stents in the treatment of malignant stenoses of the esophagus and cardia. AB - BACKGROUND AND STUDY AIMS: Several published studies have examined various self expanding metal esophageal stents for use in the palliative treatment of esophageal or cardiac neoplasia, but few have compared different self-expanding metal stents. The aim of this study was to evaluate non-covered Wallstent and Ultraflex prostheses in the treatment of malignancies in the esophagus and the cardiac region. MATERIALS AND METHODS: In a retrospective study, the effectiveness of non-covered Wallstents (46 patients) and Ultraflex stents (36 patients) was compared in the treatment of malignancies in the esophageal and cardiac regions. RESULTS: Reintervention procedures were necessary in 16 of the 46 Wallstent patients (six patients during an early phase) and in 22 of the 36 Ultraflex patients (13 during an early phase) (overall P = 0.022; early P = 0.018). The major complication in the Wallstent group was tumor ingrowth (12 of 35 complications), while in the Ultraflex group, it was incomplete deployment (18 of 49 complications). Incomplete stent deployment occurred more often in patients treated with Ultraflex (P = 0.01), and food impaction was more often observed in the Wallstent group (P = 0.001). In addition, in patients with Ultraflex stents, more complex reinterventions were necessary than those required with Wallstents (four vs. 13 complex reinterventions, P = 0.0046). Wallstents tended to improve dysphagia better than Ultraflex stents. CONCLUSION: Compared to Ultraflex stents, Wallstents have several significant short-term and long-term advantages in the palliative treatment of malignancy of the esophagus and cardia. PMID- 9201462 TI - Nonsurgical treatment of esophageal perforations after endoscopic palliation in advanced esophageal cancer. AB - BACKGROUND AND STUDY AIMS: Iatrogenic esophageal perforation during palliative endoscopic treatment in patients with incurable esophageal or cardiac cancer is a severe complication, associated with a high rate of mortality. The treatment remains controversial, since both nonsurgical and surgical treatment regimens are used. The present study describes a nonsurgical regimen. PATIENTS AND METHODS: Nine cases of perforation occurred in 142 consecutive patients referred for endoscopic palliation of dysphagia, corresponding to a perforation rate of 6%. Laser therapy was the main treatment used (argon plasma coagulation or Nd:YAG photocoagulation). RESULTS: Nonsurgical treatment was successful in six patients (75%). Two patients died (22%) as a direct result of esophageal perforation following endoscopic palliation procedures. CONCLUSION: These findings show an acceptable mortality rate using a nonsurgical treatment regimen involving broad spectrum antibiotics, nasogastric suction, and parenteral nutrition, with pleural drainage and endoprosthesis placement in addition when indicated. PMID- 9201463 TI - Colonic perforation due to colonoscopy: a retrospective study of 48 cases. AB - BACKGROUND AND STUDY AIMS: The aim of this retrospective study was to analyze data on the treatment of 48 cases of colonic perforation, with a view to defining the criteria for choosing between medical and surgical treatment. PATIENTS AND METHODS: A questionnaire requesting information about complications of colonoscopy and their treatment was sent out to four hospital gastroenterological and surgical units. RESULTS: From January 1979 to December 1993, we reviewed the records of 48 cases of colonic perforation following colonoscopy (24 perforations occurred after diagnostic colonoscopy and 24 after therapeutic colonoscopy). Diagnosis of perforation was delayed in 42% of the patients, with a mean delay of two days (range 0.5-7 days). The treatment was surgical in 35 cases, including eight in which previous medical treatment had been unsuccessful. The perforation was in the sigmoid colon in 74% of the surgical population. Operations were carried out using two procedures, including colostomy, in the case of 20 patients (57%). Colostomy closure was performed in 12 patients (60%) with no mortalities. Surgical mortality occurred in five patients (14%), in four cases due to preexisting medical diseases. Medical treatment was attempted in 21 cases, and was successful in 13, mainly in cases in which perforation had occurred after therapeutic colonoscopy (12 patients). CONCLUSION: The choice of the right type of treatment for colonoscopic perforation seems to depend on the size of the lesion. Surgical treatment is appropriate when the perforation has occurred during diagnostic colonoscopy, since the lesion in this case is usually a large colonic laceration, whereas nonsurgical treatment seems to be justified after polypectomy, as long as there is rapid clinical improvement. PMID- 9201464 TI - Endoscopic resection of submucosal tumor of the esophagus: results in 62 patients. AB - BACKGROUND AND STUDY AIMS: Although most submucosal tumors of the esophagus are benign, reliable exclusion of leiomyosarcoma requires histological analysis. However, this is rarely possible with an endoscopic forceps biopsy. In an attempt to establish the diagnosis, and as an alternative to surgery, we present here our experience with the endoscopic removal of submucosal tumors of the esophagus using two different techniques. PATIENTS AND METHODS: Sixty-two patients (38 men, 24 women, mean age 47) with submucosal tumors of the esophagus were treated endoscopically. If the tumor was less than 2 cm in diameter, polypoid, or showed a round protrusion with at least moderate elevation at endoscopy, a conventional snare polypectomy was performed. If the tumor was larger than 2 cm in diameter or only mildly elevated, the technique of modified endoscopic incisional enucleation was carried out, consisting of complete stripping of the overlying tissue followed by tumor enucleation using an electrocautery snare and a coagulation electrode. RESULTS: Based on these criteria, 36 patients underwent conventional snare polypectomy, and 25 received endoscopic incisional enucleation; complete resection of the tumor was possible in these 61 cases. In one patient, only partial removal was possible, due to firm and wide adhesions to the surrounding tissue. The tumor diameters ranged from 0.6 cm to 7.5 cm, with a mean value of 1.9 cm; 14 tumors measured more than 3 cm. At histopathology, the resected specimens were found to be 56 leiomyomas, four granular cell tumors, one neurogenic tumor, and one cyst. No serious complications such as perforation or massive bleeding occurred, and oozing bleeding, which was encountered in three patients, was easily managed by endoscopic electrocoagulation. During the follow up period (mean 38.4 months, range 3-107 months) no recurrence was observed in any of the 61 patients who received complete resections. CONCLUSION: This method of endoscopic removal of submucosal tumors of the esophagus appears to be safe and effective in experienced hands. It allows complete histopathological workup, and at the same time complete removal of the tumor. The method can be considered as an alternative to surgery in symptomatic cases. PMID- 9201465 TI - Effectiveness of local endoscopic resection of rectal carcinoid tumors. AB - BACKGROUND AND STUDY AIMS: In the treatment of rectal carcinoid tumors, confusion arises in the choice between radical surgery and local endoscopic resection, since the malignancy of individual tumors differs widely. We investigated the appropriateness of using endoscopic therapy for this disease. PATIENTS AND METHODS: Twenty-two patients were diagnosed with rectal carcinoid tumors at the First Department of Internal Medicine, Yamaguchi University School of Medicine and its affiliated hospitals, from 1977 to 1994. The tumors were resected and examined regarding their size, depth of invasion, and histological atypia. The post-treatment course in patients whose tumors were completely resected without atypia was observed by colonoscopy and ultrasonography at yearly intervals. RESULTS: In 21 patients, tumor invasion did not extend beyond the submucosal layer, and there were no signs of atypia. The size of the tumor varied from 2.2 mm to 10.0 mm in diameter, with an average of 5.4 mm. After endoscopic resection of the tumors in 18 patients and surgical local resection in three patients, no local recurrences or liver metastases were experienced. The patients survived for a minimum of 29 months and a maximum of 237 months; the mean survival period was 72.8 months. In one patient, the tumor showed cellular atypia invading into the tunica muscularis, and measured 25 mm in diameter. The patient underwent surgery, but died ten months later due to liver metastasis. CONCLUSIONS: Endoscopic treatment of rectal carcinoid tumors was found to be appropriate when the tumor measured 10 mm or less in diameter, did not infiltrate beyond the submucosal layer, and had no histological atypia. PMID- 9201466 TI - Argon plasma coagulation in flexible gastrointestinal endoscopy: pilot experiences. AB - BACKGROUND AND STUDY AIMS: In argon plasma coagulation (APC), high-frequency energy is transmitted to tissue by ionized gas, thus reducing contact with the tissue to a minimum. Successful endoscopic APC was initially reported in the palliative treatment of gastrointestinal neoplasms. The main objectives in this pilot study were to evaluate the treatment indications, efficacy and safety of the use of APC. PATIENTS AND METHODS: Between September 1994 and January 1996, APC was used to treat 125 patients with various forms of gastrointestinal pathology. RESULTS: For local palliative treatment, APC was successfully used alongside snare loop coagulation, dilation, stenting and/or radiotherapy to treat the following conditions: carcinoma of the esophagus: 15 patients, mean number of treatment sessions (MTS) 3.3; gastric carcinoma: 10 patients, MTS 4.9; rectosigmoid carcinoma: seven patients, MTS 2.7; carcinoma of the papilla of Vater: two patients, MTS 1.5. Repeated treatment was also effective for tubulovillous adenoma of the rectum (20 patients, MTS 2.5), stomach (three patients, MTS 2.0), duodenum (two patients, MTS 1.5) and papilla of Vater (two patients, MTS 3.0). In addition, APC proved helpful in coagulating the remaining tissue and achieving hemostasis after polypectomy in the colon (18 patients, MTS 1.2) and in endoscopic treatment of Zenker's diverticulum, for coagulation of the tissue bridge and hemostasis (31 patients, MTS 2.5). Finally, APC was helpful in coagulation of multiple gastric polyps (one patient, one session), hemostasis in superficial ulceration of the duodenal bulb (one patient, one session), after dilation of benign stenoses of anastomoses in the esophagus (one patient, one session) and colon (one patient, one session) and for vascular malformations in the colon (three patients, MTS 1.3), duodenum (one patient, one session), antrum (one patient, two sessions), and watermelon stomach (six patients, MTS 2.8). We recognized signs of perforation in six patients after treatment of Zenker's diverticulum (n = 3), polypectomy in the colon (n = 2) and coagulation of angiodysplasia in the cecum. Laparotomy was carried out in two patients; in one, a perforation was sutured, and in the other no focus of leakage was seen. All six patients recovered without further complications. No complications were observed in any other patients. CONCLUSIONS: These initial experiences indicate that APC seems to be effective in a number of indications, and relatively safe. Objective evaluation, a longer follow-up period, and comparative trials with other treatment modalities should follow. PMID- 9201467 TI - Clinical significance of magnetic resonance cholangiopancreatography (MRCP) compared to endoscopic retrograde cholangiopancreatography (ERCP). AB - BACKGROUND AND STUDY AIMS: The clinical importance of magnetic resonance cholangiopancreatography (MRCP) as a noninvasive diagnostic modality for investigation of the biliary tree and pancreatic duct system is under debate. Using endoscopic retrograde cholangiopancreatography (ERCP) as the gold standard, this study determined in a prospective, blinded fashion the sensitivity and further statistic values of MRCP findings for evaluation of the biliary and pancreatic tract. PATIENTS AND METHODS: Seventy-eight patients referred for ERCP were studied prospectively with MRCP and ERCP during a 12-month period. All images were interpreted on a blinded basis by two radiologists. Any dilations, strictures, and intraductal abnormalities were recorded and correlated with the clinical diagnoses. RESULTS: MRCP images of diagnostic quality were obtained in 76 of the 78 patients (97%). Magnetic resonance cholangiography (MRC) showed sensitivities (and positive predictive values) of 71% (62%) for recognition of normal bile ducts, 83% (91%) for recognition of dilation, 85% (100%) for recognition of strictures, 77% (91%) for correct stricture location, and 80% (100%) for diagnosing bile duct calculi. In addition, the sensitivity of MRC in classifying benign and malignant strictures was 50% and 80%, respectively. The statistical values (sensitivity and positive predictive value) for magnetic resonance pancreatography findings were determined for the recognition of normal pancreatic ducts (33% and 50%), recognition of dilation (62% and 100%), recognition of strictures (76% and 87%) and correct location (66% and 100%), diagnosis of benign strictures (87% and 87%) and malignant strictures (60% and 75%), and for diagnosing pancreatic duct stones (60% and 100%). CONCLUSIONS: MRCP is capable of providing diagnostic information equivalent to ERCP in many patients, and should be applied whenever established techniques provide no results, or inadequate results. PMID- 9201468 TI - Preliminary experience with a new ultraviolet-cured expandable plastic endoprosthesis in the management of malignant esophageal strictures. AB - BACKGROUND AND STUDY AIMS: Different types of expandable metal stent are currently available for the palliative treatment of malignant esophageal strictures. To overcome some of the disadvantages involved in the design of metal mesh stents, we designed a balloon-expanded plastic endoprosthesis, which is hardened by irradiation with ultraviolet light after deployment. We present here our preliminary results. PATIENTS AND METHODS: From April 1995 to January 1996, four patients with unresectable esophageal malignancies were treated with this stent. Insertion of the stent was the only palliative procedure carried out. The patients were followed up until death. RESULTS: Stent placement was successful in all patients, and no procedure-related complications occurred. Dysphagia was reduced from an average score of 2.6 to 1.0. Early complications included retrosternal pain of limited duration in one patient. One patient died 72 hours after uncomplicated stent placement, due to cardiac arrhythmia. Late complications were limited to tumor overgrowth in two patients, after a mean of 104 days. The stent patency rate averaged 92 days. CONCLUSIONS: In patients with a malignant esophageal stricture, this newly developed expandable endoprosthesis is effective in relieving dysphagia. The deployment of the stent is easy and safe. The endoprosthesis has potential advantage over current expandable metal mesh stents. PMID- 9201469 TI - Endoscopic resection of gastrointestinal tumors: how far can the endoscopist go? PMID- 9201470 TI - Argon plasma coagulation (APC): ballyhoo or breakthrough? PMID- 9201471 TI - Transmission of hepatitis C and prion diseases through digestive endoscopy: evaluation of risk and recommended practices. PMID- 9201472 TI - ESGE guidelines for the prevention of endoscopic transmission of type C hepatitis and update on Creutzfeldt-Jakob disease. European Society of Gastrointestinal Endoscopy. PMID- 9201473 TI - Endoscopic sewing and stapling machines. PMID- 9201474 TI - Nitinol self-expanding stents: when severe twisting occurs, self-expansion may ensue after a waiting period. AB - One of the major disadvantages of nitinol endocoil stents is said to be incomplete deployment leading to twisting and stent dysfunction. We report a patient with irresectable pancreatic head cancer where severe twisting of the nitinol stent occurred which resolved on the day after stent insertion; the stent took its normal shape ten days later, whereafter the patient lived without stent related problems for one year until his death. This shows that prospective management rather than immediate stent extraction may be warranted in such cases of stent dysfunction. PMID- 9201475 TI - Endosonography-guided endoscopic resection of duodenal carcinoid tumor. AB - Endoscopic resection techniques using snare polypectomy with or without submucosal saline injection have also been applied to resect smaller duodenal carcinoid tumors. We report on two patients where endoluminal ultrasound using a small diameter probe was used to visualize the adherence of the carcinoids to the underlying wall layers and the separating effect of subtumoril saline injection. One patient, in whom a clear separation between the tumor and the underlying tissue after saline injection was visualized, underwent successful endoscopic resection. In the other patient no clear separation could be achieved after saline injection and the patient underwent surgical removal of his tumor; both tumors were 1 cm or less in their maximal diameter. PMID- 9201476 TI - Carcinoid tumors of the duodenum: report of three cases treated by endoscopic resection. AB - Three patients with small (less than 1 cm) duodenal carcinoid tumors are described, two of whom underwent endoscopic resection using simple snare polypectomy. In the third patient, a small tumor (5 mm) could no longer be found after initial biopsies on subsequent repeated endoscopic-bioptic follow-up. All three patients are free of local recurrence or systemic symptoms 2-14 years after the procedure. PMID- 9201477 TI - Microcalculi of the common bile duct: a misdiagnosed problem. PMID- 9201479 TI - Reflux esophagitis: a complication of Helicobacter pylori eradication therapy? PMID- 9201478 TI - Endoscopic diagnosis of postoperative ileocolonic intussusception. PMID- 9201480 TI - Laparoscopic cholecystectomy is a safe procedure for the treatment of porcelain gallbladder. PMID- 9201481 TI - Laparoscopic management of cholecystogastric fistula. PMID- 9201482 TI - Endoscopy-assisted laparoscopic resection of the gastric wall facilitated by a double-lifting method. PMID- 9201483 TI - Laparoscopic cecoplication for mobile cecum. PMID- 9201484 TI - A new option in the treatment of complete and acute obstruction due to colorectal cancer. PMID- 9201485 TI - Endoscopic appearance of primary anorectal melanoma. PMID- 9201486 TI - Unusual rectal foreign body: treatment using argon-beam coagulation. PMID- 9201487 TI - Asymptomatic pneumoperitoneum after endoscopic treatment of pseudocysts. PMID- 9201488 TI - Ultrasound-guided emergency endoscopic retrograde biliary drainage without radiography. PMID- 9201489 TI - Reloading a variceal rubber band ligator with hemorrhoidal bands: an inexpensive and effective method. PMID- 9201490 TI - Developing primary care through education. PMID- 9201491 TI - 'Ending up a GP': a qualitative study of junior doctors' perceptions of general practice as a career. AB - OBJECTIVE: This study aimed to investigate junior doctors' perception of general practice as a career. METHOD: In-depth interviews with a purposive sample of 54 junior doctors were carried out. RESULTS: Three main criteria were identified: clinical content of practice, lifestyle, and organizational context of practice. Clinical content was most highly valued, but was recognized to conflict with lifestyle. Compared with hospital medicine, general practice was associated with inferior clinical content but superior lifestyle. Views on organizational context were more equivocal. Choice of general practice as a career was often based on negative judgements. CONCLUSIONS: Junior doctors' perceptions of general practice are expressions of a hospital-centric culture. Criteria for career choice are diffuse and complex. There was no evidence that the 1990 GP Contract deterred recruitment. The compromise between intrinsic satisfaction and lifestyle may be significant for GP morale. PMID- 9201492 TI - Does a computerized price comparison module reduce prescribing costs in general practice? AB - OBJECTIVE: We aimed to assess the trends in prescribed defined daily doses (DDD) and drug expenses before and after the introduction of a computerized cost containment module into the computer record system of a defined group of GPs. The GPs' expectations for and experiences with the module were examined. METHOD: We performed a controlled follow-up study on antecedent data before and after intervention. A questionnaire was administered to the intervention group at the introduction and 1 year later. Data on prescribing were collected in the database of the Health Insurance Aarhus County, as a normal routine for accounting. The GPs were not aware of the ongoing cost supervision study. Additional cost information software was introduced on 1 January 1993 to 20 practices with 28 GPs. The software assisted the GPs in a semiautomatic way to identify and prescribe the cheapest drugs. The subjects comprised 158 practices including 231 GPs in Aarhus County, Denmark. Questionnaires were sent to the 20 intervention practices. The main outcome measures were prescribed DDD, reimbursement for prescribed drugs, and reimbursement per prescribed DDD quarterly during 1992 and 1993. RESULTS: Compared with the controls there were no changes in prescribed DDD, reimbursement for prescribed drugs, and reimbursement per prescribed DDD in the intervention group after the introduction of the module. CONCLUSION: Simply giving a random group of GPs computer assistance to choose less expensive drugs did not reduce expenditure per DDD. Cost containment procedures should be more intensive than just giving the doctors a computer-assisted decision aid. PMID- 9201493 TI - Identifying relevant diagnostic studies in MEDLINE. The diagnostic value of the erythrocyte sedimentation rate (ESR) and dipstick as an example. AB - OBJECTIVE: We aimed to examine sensitivity and positive predictive value of MEDLINE searching for diagnostic studies, relevant for the primary health care setting. METHOD: Results of MEDLINE searches were compared with a reference standard collection of studies on two subjects, the diagnostic value of ESR in discriminating between 'pathology' and 'no pathology', and the dipstick method in diagnosing urinary tract infections. The main outcome measures were sensitivity (proportion of the total number of reference standard diagnostic studies that could be identified by the search) and positive predictive value (proportion of the total number of publications retrieved by MEDLINE that were incorporated in the reference standard). RESULTS: The combined MeSH and freetext search was more sensitive than MeSH term searching only, for both the ESR and the dipstick search. With this combined search sensitivities of 0.91 and 0.98 and predictive values of 0.10 and 0.68 were found for ESR and dipstick respectively. By restricting the search with keywords describing the primary health care setting the predictive values increased to 0.72 and 1.00 but sensitivity dropped to 0.10 and 0.07 (ESR and dipstick respectively). CONCLUSION: Combining freetext and MeSH term searching, without restriction to the primary health care setting, is a valuable strategy in systematically searching for available evidence on the value of a diagnostic test in the scope of a specific disease. The predictive value seems to depend on the breadth of the disease area. MEDLINE should provide a term such as 'diagnostic evaluation study' to be used in the limit field Publication Type to specify diagnostic studies. PMID- 9201494 TI - Impact of local evidence-based clinical guidelines--a Danish intervention study. AB - OBJECTIVE: We aimed to evaluate whether a cheap and less labour-intensive regional implementation strategy for guidelines was sufficient to change knowledge and behaviour among GPs. The model studied was the implementation of anticoagulant therapy to prevent stroke in atrial fibrillation. METHOD: The intervention took place in the county of Viborg (149 GPs), Denmark, with the county of Ringkobing (166 GPs) as control. A local interdisciplinary steering group modified national college-based guidelines, followed by a regional dissemination and implementation strategy. The effect of the intervention was evaluated during a follow-up period by a repeated questionnaire and by monitoring prescriptions for oral anticoagulants in 1993 and 1995 in the Danish National Health Service. RESULTS: Adherence to the guidelines was higher after the intervention but, considering secular trends and baseline differences, the guidelines had no significant effect. The use of oral anticoagulants increased substantially in both counties during the 2-year follow-up period, but the difference in relative change between the counties was negligible. Adherence to the guidelines could not be predicted by any of the reported practice characteristics or attitudes to guidelines. CONCLUSION: Despite solid scientific documentation and regional modification to establish ownership of nationally agreed guidelines, the impact of guidelines on GPs' knowledge and behaviour was disappointing. PMID- 9201496 TI - Preventive care for patients following myocardial infarction. The Wessex Research Network (WReN). AB - OBJECTIVE: We aimed to assess general practice care for patients following a myocardial infarction (MI). METHOD: A structured review was carried out of general practice records of patients identified from hospital administration data. A total of 266 survivors following MI were identified from the discharge data of 13 hospitals in Southern England and registered with 71 GPs belonging to the Wessex Research Network. Median time since hospital discharge was 2.1 years. The main outcome measures were the provision of appropriate preventive care, including cardiac rehabilitation, drug therapy, and lifestyle advice for modifiable risk factors. RESULTS: Basic care was provided to nearly all patients; 253 (95.1%, 95% Cl 91.8-97.4) had blood pressure documented after their MI, 216 of 234 patients eligible for aspirin (92.3%; 88.1-95.4) had been recommended treatment, and the provision of advice on smoking cessation was documented for 27 of 33 continuing smokers (81.8%; 64.5-93.0). However, only 73 of 236 patients eligible to attend a structured rehabilitation programme (30.9%; 25.0-36.8) were documented as having received rehabilitation. Of 89 patients with heart failure following MI, 33 (37.1%; 27.1-48.0) had no record of having been offered treatment with an ACE inhibitor. Total cholesterol measurement was documented for only 144 patients (54.1%; 48.1-60.1). We estimate that there is still the potential to prevent between 4 and 9 deaths in this group of 266 surviving patients in the next 2 years by further improving the quality of follow-up care. CONCLUSIONS: Preventive care in patients with proven ischaemic heart disease in general practice remains haphazard, even among doctors enthusiastic to participate in research and to audit their quality of care. As general practitioners we should ensure that we are providing high quality preventive care to patients with clinical disease before we focus on the even more demanding task of primary prevention. PMID- 9201495 TI - Doctors' willingness to refer elderly patients for elective surgery. AB - OBJECTIVES: We aimed to examine the relationship between doctors' willingness to refer elderly patients for elective surgical operations and patients' age, comorbidity, institutionalization, living habits and signs of dementia. METHOD: A random selection of 837 medical doctors in Finland (response rate 56%) received a postal questionnaire consisting of 18 vignettes, i.e. imaginary patient cases. Respondents were asked whether they would refer the patient on the vignette for elective surgical operation, treat the patient conservatively, or choose some other alternative. In the vignettes, the age of patients was randomly varied between 65 and 85, at 5-year intervals, to provide eight different questionnaires, and each respondent obtained one of them. RESULTS: The proportion of doctors willing to refer the patients for surgery was inversely related to the patients' age: in all the vignettes, doctors said they would refer fewer patients in the oldest age groups. Almost all the doctors claimed they would refer healthy, home-dwelling persons aged 65-70 years for operations. In the oldest age groups of patients, the doctors' willingness to refer was highest for cataract operations (69%) and hip prosthesis operations (63%), but only 18% of doctors would refer such patients for coronary by-pass operations. Comorbidity and institutionalization were associated with fewer doctors referring the patients: the proportion of doctors willing to refer these patients was about half that of those willing to refer otherwise healthy and home-dwelling patients. Smoking by patients also decreased the proportions of doctors willing to refer, but moderate signs of dementia in an elderly patient with cataract were associated with only a slight decrease in referring. CONCLUSIONS: Doctors are less willing to refer old patients for elective surgery, but comorbidity, patients' lifestyle and institutionalization have a greater effect on referrals than age. PMID- 9201497 TI - Adult asthma review in general practice: nurses' perception of their role. AB - BACKGROUND: Asthma clinics have become widespread in general practice with nurses now playing an important role in asthma review. However, little is known about training of nurses carrying out reviews and how this affects the nurse role in patient management. OBJECTIVES: We aimed to discover the level of asthma training of practice nurses carrying out review of adult asthma patients in one Health Authority and to see if this has any effect on their perception of their role. METHOD: All 187 practice nurses in Grampian were sent a postal questionnaire investigating how asthma review is organized in general practice, their role in review and the asthma training they had received. Personal interviews were carried out with 17 nurses, exploring in more depth the topics covered in the questionnaire. RESULTS: A total of 167 nurses from 92% of the practices in Grampian responded, of whom 61% carried out asthma reviews. Among nurses carrying out reviews 71% did so on their own. 49% of nurses had or were training for advanced asthma qualification. Nurses without an asthma qualification were significantly more likely to feel that their training was not sufficient for their asthma related tasks (54% versus 11%, P = 0.0002). Nurses without advanced asthma qualifications were less likely to provide or review a self-management plan (29% versus 49%, P = 0.01), to review patient PEF recording (38% versus 65%, P < 0.01), to discuss patient worries (75% versus 94%, P < 0.05) or to make the initial diagnosis of asthma (24% versus 76%, P < 0.005). Nurses were unlikely to view their role as fully responsible unless they had an asthma qualification (13% versus 49%, P < 0.001). CONCLUSION: Nurses without advanced asthma qualifications do not feel fully confident in responsibility for patient management. Nurses without training are more likely to only carry out routine monitoring at reviews while nurses with asthma training are more likely to actively develop patient self-management skills. This suggests that nurses should be supported to obtain asthma qualifications if they are to give the best possible care to asthma patients. PMID- 9201498 TI - Refusal of videorecording: what factors may influence patient consent? AB - BACKGROUND: Videorecording of the consultation has become a widespread training and research technique in general practice. Previous studies have suggested a good level of patient acceptability. OBJECTIVE: This study aimed to elucidate patient factors associated with refusal to be videoed. METHOD: A research study of GP detection of psychological problems was carried out by opportunistic recruitment of adult attenders in primary care for collection of sociodemographic data, mental health status, and video of consultation. Patients were consented for participation in a research project by a skilled research assistant while waiting to see their general practitioner. This involved a two-stage process, where completion of a brief questionnaire was followed by consent for video before the patient entered the consulting room. RESULTS: Although the overall response rate was 85%, a surprisingly high level of refusal of consent by patients for their consultations to be videoed was found. The data were therefore analysed to examine the characteristics of the patient population, and to look for possible associated factors. The main factors associated with refusal of video consent were decreasing age, and overt presentation of a psychological problem. Although patients who were defined (either in the opinion of the GP or by mental health questionnaire score) as psychologically distressed were more likely to decline to be videoed, this was a less significant predictor of whether the patient would consent. CONCLUSIONS: Consent to be videoed may be more context specific than previous studies have suggested. Refusal to be videoed appears to be associated with psychological problems, but the relationship is complex, and sociodemographic factors also play a part. The factors which influence consent, including the reason given for recording, need further clarification. PMID- 9201499 TI - Awareness of diagnostic and clinical features of fibromyalgia among family physicians. AB - OBJECTIVES: The aim was to assess the awareness and knowledge of family physicians about diagnostic and clinical features of fibromyalgia syndrome (FS), and to evaluate the contribution of rheumatology education to the improvement of this knowledge. METHODS: A detailed questionnaire on FS was completed by 172 family physicians. A composite score, based on five items, was constructed to quantitatively assess knowledge of FS (maximum score of 5). A comparison was made between physicians exposed to extensive education on FS (in Beer Sheva medical centre) and physicians without such exposure (in other centres). RESULTS: Ninety six per cent of the physicians claimed to be familiar with FS. They recognized most of the FS-related symptoms, but had limited knowledge of the diagnostic criteria, treatment modalities and prognosis. Only 55% knew that FS is associated with widespread pain and 25% were familiar with the point count criterion. Physicians trained in Beer Sheva scored significantly higher than those trained elsewhere: 3.0 +/- 1.2 versus 2.4 +/- 1.2, respectively (P = 0.006), and their knowledge of FS treatment was significantly better. CONCLUSION: Family physicians in Israel are quite unfamiliar with the diagnostic criteria of FS, though educational exposure improves their awareness and knowledge. PMID- 9201500 TI - Biopsychosocial features of patients with widespread chronic musculoskeletal pain in family medicine clinics. AB - OBJECTIVES: We aimed to describe the clinical and psycho-socio-familial features of patients with widespread chronic musculoskeletal pain (WCMP)/fibromyalgia (FM) in primary care settings. To detect differences and similarities between both 'entities'. METHODS: An observational study was carried out with a newcoming clinical case series. Five family medicine surgeries were included. Patients aged from 18 to 50 attended the clinic to fulfil pain criteria for WCMP. Differences between WCMP and FM were based on the presence of 'tender points'. Measurements were made of general characteristics, occupation, pain description, symptoms, tender points, radiographic and laboratory studies, and questionnaires to assess self-rated health (NHP), social support (DUKE), family support (Family-APGAR), and psychopathological traits (CAQ). RESULTS: We identified 48 patients (23 WCMP;25 FM) with a mean age of 38.4 +/- 8.4; 95.8% were females. The back was the anatomical place most frequently reported (93.7%) and 34.8% of the patients pointed out the nape as being the most painful place. The average duration of pain was 6.7 +/- 7 years. Unsteadiness (72.9%), impairment in symptoms with weather (70.8%), with activity (70.8%) and general fatigue (68.8%) were the most frequently detected symptoms. 'Pain' (59.5) and 'energy' (54.4) were the scales of the NHP test most affected. Half of the patients were poorly satisfied with the responses of their families to their needs and over 60% showed psychopathological traits. The patients with FM reported worse self-rated health than those with WCMP; the number of years of pain (4.9 versus 8.2) and the number of symptoms (6.6 versus 8.9) were both greater in patients with FM. CONCLUSIONS: The clinical and psychological features of patients with WCMP-FM are similar to those reported by others. The self-rated health reported by these patients is poor and closer to that reported by patients suffering other chronic osteoarticular diseases. These results support the hypothesis that FM should be considered as more advanced clinical stage of the widespread musculoskeletal pain continuum. PMID- 9201501 TI - 'It would be good to know you're not alone': the health care needs of women with menstrual symptoms. AB - OBJECTIVES: We aimed to explore how menstrual symptoms affect women, women's health care needs, and their expectations and experiences when seeking care; to identify ways to assist women in having their needs met. METHODS: Qualitative research using focus group methodology was carried out. Identification of women experiencing menstrual symptoms through a random community survey of 200 women aged 30-50 years in the Hunter region of New South Wales. Focus group discussions were recorded and transcripts were analysed. RESULTS: All women attending focus groups had sought medical advice for their menstrual problems. Having one doctor with whom they felt comfortable was important. Women expressed difficulties asking questions and were concerned that symptoms may not be taken seriously or may be dismissed as psychological. There was widespread acceptance of alternative 'natural' therapies. CONCLUSIONS: Interactions between doctors (particularly GPs) and women with menstrual symptoms are central to how women perceive the care they receive. There is a need for doctors to demonstrate empathy. For many women, what they needed most from their doctors was to be understood and 'to know they weren't alone'. PMID- 9201502 TI - Access to medical care one year prior to diagnosis in 100 HIV-positive women. AB - BACKGROUND: Anonymous antenatal testing for HIV antibodies suggests that the majority of HIV-positive women in the UK remain undiagnosed. Primary care reaches the majority of the population and women present more often than men. Sexual health matters are frequently raised, so there is an opportunity to discuss concerns with respect to HIV. Women often present with advanced HIV disease and with antiviral treatments proving to be more effective, there is now an even greater incentive to diagnose HIV early. OBJECTIVE: We aimed to look at access to medical care 1 year prior to HIV diagnosis in 100 HIV-positive women and to establish whether a discussion regarding HIV was recalled. METHOD: The setting was an established clinic for HIV-positive women in the Royal Free Hospital, London. In a 6-month period a questionnaire was completed by 100 women with their clinic doctor. RESULTS: Women were young (mean age 31). Most (84%) were infected by heterosexual sex. Forty-six per cent of the women presented with symptomatic HIV or AIDS and 50% were black Africans, hence there is a large ethnic bias in this sample. General practice was accessed by 65% of the women 1 year prior to HIV diagnosis. Few (14%) women recalled a discussion concerning HIV. Secondary care settings such as gynaecology and general outpatients were also frequently attended, but again HIV was apparently rarely discussed. There was no significant difference when variables such as time since diagnosis, health care setting, or ethnic group were concerned with respect to recall of a discussion concerning HIV. CONCLUSIONS: Despite coming from 'high risk' groups or presenting with symptomatic disease in the year prior to HIV diagnosis, few women recalled discussing HIV in either primary or secondary care settings. As these sites were commonly accessed, we feel that doctors and other health care workers should be encouraged and trained to raise HIV more routinely in their consultations. PMID- 9201503 TI - Selections from current literature: obsessive compulsive disorder. PMID- 9201504 TI - Distribution and viability of cultured human chondrocytes in a three-dimensional matrix as assessed by confocal laser scan microscopy. PMID- 9201505 TI - Sex related differences in the adhesion, migration, and growth of rat aortic smooth muscle cells in culture. PMID- 9201506 TI - Application of surfactin for mycoplasma inactivation in virus stocks. PMID- 9201507 TI - Chick embryo brain cultures enriched for neurons or astroglial cells support the replication of influenza A, B, and C viruses. PMID- 9201508 TI - Initiation of epithelial cell cultures from palleal buds of Botryllus schlosseri, a colonial tunicate. PMID- 9201509 TI - A cell line from the gill tissues of Indian cyprinoid Labeo rohita. PMID- 9201510 TI - Adult rat cardiomyocyte proliferation assay. PMID- 9201511 TI - Three-dimensional endothelial-tumor epithelial cell interactions in human cervical cancers. AB - The purpose of this study is to understand the multicellular interaction between tumor epithelial (TEC) and human umbilical vein endothelial cells (HUVEC). The development of in vitro systems in which to coculture these cells as multicellular aggregates is very critical. Cell lines were established from cervical tumor cells (n = 6) and two from HUVEC (n = 2) and they were cultured as three-dimensional (3-D) multicellular-cultures using Cytodex-3 microcarrier beads in the rotating wall vessel (RWV). After a 240-h incubation, TEC and HUVEC proliferated exponentially to 4.2 x 10(7) and 2.2 x 10(7) cells/ml, respectively, without requiring a feeder layer; in contrast to the two-dimensional (2-D) cultures that average about 8 x 10(5) cells/ml. Phase contrast microscopy indicated formation of 3-D aggregates that varied in size from 0.5 to 5 mm. The size of the aggregates (1-5 mm, 6-14 microcarriers) increased over time; however, the number of aggregates (0.5-1 mm, 2-5 microcarriers) decreased over a long-term incubation (240 h) because the cells merged to form large clumps. Maximum aggregation was observed with TEC at 120 h and HUVEC at 96 h. The culture of TEC in the absence of HUVEC produced minimal differentiation in contrast to cocultures. The TEC and HUVEC as cocultures in RWV proliferated at an accelerated rate (1.3 x 10(7) cells/ml, 96 h). The TEC-HUVEC coculture presented tubular structures penetrating the tumor cell masses, forming aggregates larger in size than the monocultures and typically with greater cell mass and number. The cells were viable (trypan blue exclusion) and metabolically active (glucose utilization) until 240 h. These data suggest that RWV provides a new model that allows us to investigate the regulatory factors that govern tumor angiogenesis. PMID- 9201512 TI - TGF-beta 1 reverses PDGF-stimulated migration of human aortic smooth muscle cells in vitro. AB - Platelet-derived growth factor (PDGF) and transforming growth factor beta-1(TGF beta 1) were tested separately or together for the ability to stimulate migration of human aortic vascular smooth muscle cells (VSMC). PDGF (10 ng/ml) stimulated migration of VSMC over a 48-h period. TGF-beta 1 (10 ng/ml) had no effect on migration during the same period. VSMC exposed simultaneously to both TGF-beta 1 and PDGF exhibited diminished migration (50%) when compared to cells treated only with PDGF. Cells that migrated in the presence of PDGF possessed short actin cables that extended from cellular processes at the leading edge of migrating cells; focal adhesions containing the alpha v beta 3/beta 5 integrins localised to the same region. Cells grown in the presence of TGF-beta 1 exhibited long, intensely stained actin filaments that spanned the entire length of the cell and were similar to untreated control VSMC. Focal adhesions containing alpha v beta 3/beta 5 distributed evenly on the basal surface in both TGF-beta 1-treated cells and control cultures. Cellular responses to PDGF were mitigated when TGF-beta 1 was present in the culture medium. VSMC grown in the presence of both PDGF and TGF-beta 1 exhibited elongated actin filaments that were similar to nonmotile TGF beta 1-treated cultures. Concomitant exposure of VSMC to PDGF and TGF-beta 1 resulted in focal adhesions that distributed evenly on the lower cell surface. This study suggests that TGF-beta 1 can partially reverse the stimulatory effect of PDGF on VSMC migration in vitro by modifying the actin cytoskeleton and the distribution of the alpha v beta3/beta 5 integrins. PMID- 9201513 TI - Improved conditions for culture of biosynthetically active cockroach corpora allata. AB - Currently, short-term culture of insect corpora allata is most often performed in TC199. We now show that L-15B, a medium widely used in arthropod tissue culture, is superior to TC199 for both short- and long-term culture of cockroach corpora allata. In 3-h and 48-h incubations, juvenile hormone biosynthesis by corpora allata from Diploptera punctata was significantly higher in L-15B than in TC199. In addition, in both media, corpora allata activity was significantly improved by flotation of glands at the medium surface. Characteristics of L-15B responsible for its superiority were examined by comparison of gland activities in several TC199 formulations that had been modified in different ways to be more similar to L-15B. Adjusting the osmotic pressure of TC199 (288 mOsm/l) to near that of L-15B (362 mOsm/l) and D. punctata hemolymph (360 mOsm/l) significantly improved gland activity during the second 12 h of a 36-h incubation. Increasing the concentrations of amino acids, sugars, and organic acids in TC199 to the same levels as in L-15B significantly improved gland activity during both the second and third 12-h intervals of a 36-h incubation. These results suggest that L-15B is superior to TC199 because L-15B is isoosmotic with D. punctata hemolymph and because L-15B, like cockroach hemolymph, contains a high level of organic constituents. It is therefore more appropriate to use L-15B than TC199 for short term in vitro assays of juvenile hormone biosynthesis and for extended corpora allata culture. PMID- 9201514 TI - Three-dimensional growth patterns of various human tumor cell lines in simulated microgravity of a NASA bioreactor. AB - Growth patterns of a number of human tumor cell lines that from three-dimensional structures of various architectures when cultured without carrier beads in a NASA rotary cell culture system are described and illustrated. The culture system, which was designed to mimic microgravity, maintained cells in suspension under very low-shear stress throughout culture. Spheroid (particulate) production occurred within a few hours after culture was started, and spheroids increased in size by cell division and fusion of small spheroids, usually stabilizing at a spheroid diameter of about 0.5 mm. Architecture of spheroids varied with cell type. Cellular interactions that occurred in spheroids resulted in conformation and shape changes of cells, and some cell lines produced complex, epithelial-like architectures. Expression of the cell adhesion molecules, CD44 and E cadherin, was upregulated in the three-dimensional constructs. Coculture of fibroblast spheroids with PC3 prostate cancer cells induced tenascin expression by the fibroblasts underlying the adherent prostate epithelial cells. Invasion of the fibroblast spheroids by the malignant epithelium was also demonstrated. PMID- 9201515 TI - Extended primary culture of human hepatocytes in a collagen gel sandwich system. AB - To develop a strategy for extended primary culture of human hepatocytes, we placed human hepatocytes between two layers of collagen gel, called a "collagen gel sandwich." Maintenance of hepatocellular functions in this system was compared with that of identical hepatocyte preparations cultured on dry-collagen coated dishes or cocultured with rat liver epithelial cells. Human hepatocytes in a collagen gel sandwich (five separate cultures) survived for more than 4 wk, with the longest period of culture being 78 d. They maintained polygonal morphology with bile canaliculuslike structures and high levels of albumin secretion throughout the period of culture. In contrast, hepatocytes on dry collagen became feature-less, and albumin secretion could not be detected after 14 d of culture. This loss of albumin secretion was partially recovered by overlaying one layer of collagen gel. Ethoxyresorufin O-deethylase activity, associated with cytochrome P450 1A2, was detected basally up to 29 d in collagen gel sandwich culture. These activities were induced four- to eightfold after induction with dibenz(a,h)anthracene. Cocultures also maintained basal activity up to 29 d. However, their inducibility was lower than that of hepatocytes in collagen gel sandwich. No ethoxyresorufin O-deethylase activity was detected in hepatocytes cultured on dry-collagen at 7 d. Thus, the collagen gel sandwich system preserves differentiated morphology and functions of human hepatocytes in primary culture for a prolonged period of time. This system is a promising model for studying human hepatocellular function, including protein synthesis and drug metabolism in vitro. PMID- 9201516 TI - Individual and combined effects of calciotropic hormones and growth factors on mineral metabolism in embryonic chick tibiae. AB - We have investigated single and combined effects of calciotropic hormones and growth factors on the regulation of alkaline phosphatase (ALP) activity and calcium metabolism in an optimized serum-free bone organ culture system of embryonic chick tibiae. Parathyroid hormone PTH(1-34) alone mobilized calcium from bone tissue time- and dose-dependently and inhibited ALP activity. Both the bisphosphonate (BM 21.0955) and to a lesser extent salmon calcitonin alone slightly increased calcium uptake and inhibited the stimulation of bone resorption by PTH(1-34). 1,25(OH)2D3 mobilized calcium and inhibited ALP activity in contrast to 24,25(OH)2D3 which inhibited ALP activity but had no significant effect on calcium metabolism. Interestingly the combination of PTH(1-34) with 1,25(OH)2D3 but not 24,25(OH)2D3 reduced calcium mobilization. The combination of the midregional fragment PTH(28-48), which by itself has no effect on calcium metabolism, with 1,25(OH)2D3 reduced calcium mobilization more efficiently. Several PTH-regulated mediators have been assayed in this system. Of the tested growth factors, IGF-I at high concentrations caused bone resorption with no effect on ALP activity. TGF-beta 1 (transforming growth factor beta) and BMP-2 had no significant effect on calcium metabolism; however, ALP activity was inhibited by TGF-beta 1 and induced dose dependently by BMP-2. Of the other factors known to be present in bone, platelet-derived growth factor (PDGFA/B) and epidermal growth factor (EGF) had a small effect on calcium mobilization but had no effect on ALP activity. bFGF reduced ALP activity slightly without an effect on calcium metabolism. Our results show that this in vitro system can mimic some interactions of calciotropic hormones in vivo and allows the assaying of mediators in terms of regulation of ALP activity and of calcium metabolism. PMID- 9201517 TI - Differential requirements of two insect cell lines for growth in serum-free medium. AB - The development of a serum-free medium that supports the growth of cells from a Spodoptera frugiperda and a Lymantria dispar cell line is reported. A yeast hydrolysate provided the B-vitamin complex, and a combination of a meat hydrolysate and tryptose provided most of the free amino acids required for cell growth. Supplemental cystine and methionine were required to achieve maximum cell growth. The serum or serum replacements used in earlier formulations were replaced with commercial lipid preparations and increased levels of iron salts. Although the cell growth cycle had a somewhat extended lag phase and the population doubling time of the S. frugiperda cells was longer than on serum containing medium, the saturation densities were much higher. Spodoptera cells grown in this medium replicated the Autographa californica nuclear polyhedrosis virus well, producing 8.71 x 10(6) TCID50 extracellular virus and 4.4 x 10(6) polyhedra/ml culture. The specific activity of the polyhedra was somewhat less than that of polyhedra produced in insects. PMID- 9201518 TI - Neutrophil adhesion molecules and MOF. PMID- 9201519 TI - Interpreting dual-sugar absorption studies in critically ill patients: what are the implications of apparent increases in intestinal permeability to hydrophilic solutes? PMID- 9201520 TI - The hemodynamic consequences of mechanical ventilation: an evolving story. PMID- 9201521 TI - Base deficit after major trauma directly relates to neutrophil CD11b expression: a proposed mechanism of shock-induced organ injury. AB - OBJECTIVE: To determine whether expression of neutrophil integrin receptors is related to the degree of post-traumatic shock. DESIGN: Data were collected prospectively on patients with major trauma admitted to the surgical intensive care unit. SETTING: Denver General Hospital, Colorado. PATIENTS AND PARTICIPANTS: 17 severely injured adults. MEASUREMENTS AND RESULTS: The mean fluorescence intensity and per cent positive of neutrophil integrin receptors CD11 b, CD18 and CD11 a, and systolic blood pressure, blood transfusion, lactate and base deficit as indices of shock. CD11 b expression on circulating neutrophils was increased 6 and 12 h after trauma. After correcting for the other shock indices, base deficit predicted CD11 b expression at 12 h. CD11 b expression was negatively correlated with the circulating neutrophil count. CONCLUSIONS: The degree of metabolic acidosis after trauma correlates directly with CD11 b receptor expression on circulating neutrophils. This relation may be the mechanism whereby post traumatic shock results in neutrophil sequestration and neutrophil-mediated organ injury and failure. PMID- 9201522 TI - Gastrointestinal permeability following cardiopulmonary bypass: a randomised study comparing the effects of dopamine and dopexamine. AB - OBJECTIVE: To compare the effects of dopexamine and dopamine on the mucosal permeability of the gastrointestinal tract (GIT). DESIGN: Prospective, randomised clinical trial. SETTING: Intensive care unit of a postgraduate teaching hospital, London, England. PATIENTS: Thirty patients undergoing elective surgery involving cardiopulmonary bypass, performed by a single surgeon. INTERVENTIONS: Patients were randomly assigned to receive either dopexamine 2.0 micrograms/kg per min or dopamine 2.5 micrograms/kg per min for the duration of the study period. MEASUREMENTS AND MAIN RESULTS: Hemodynamic parameters and gastric intramucosal pH (pHi) were measured at intervals throughout the study. GIT permeability was measured once, post-operatively, using the ratio of absorbed lactulose to L rhamnose. The groups were similar with respect to demographics, pre- and post operative risk factors. The lactulose/rhamnose ratio was (mean +/- SEM) 0.44 +/- 0.10 in the dopexamine group vs 0.65 +/- 0.08 in that receiving dopamine (p < 0.05). The dopexamine group had a significantly higher oxygen delivery preoperatively (479.5 +/- 32.0 ml/min per m2 vs 344.4 +/- 23.9 ml/min per m2 for dopamine, p < 0.01), but no other significant differences emerged between the groups. CONCLUSIONS: Compared to dopamine, dopexamine reduces GIT permeability following surgery involving cardiopulmonary bypass. The mechanism of this effect remains unclear. PMID- 9201523 TI - Saccharomyces boulardii prevents diarrhea in critically ill tube-fed patients. A multicenter, randomized, double-blind placebo-controlled trial. AB - OBJECTIVE: To assess the preventive effect of Saccharomyces boulardii on diarrhea in critically ill tube-fed patients and to evaluate risk factors for diarrhea. DESIGN: Prospective, multicenter, randomized, double-blind placebo-controlled study. SETTING: Eleven intensive care units in teaching and general hospitals. PATIENTS: Critically ill patients whose need for enteral nutrition was expected to exceed 6 days. INTERVENTION: S. boulardii 500 mg four times a day versus placebo. MEASUREMENTS AND RESULTS: Diarrhea was defined by a semiquantitative score based on the volume and consistency of stools. A total of 128 patients were studied, 64 in each group. Treatment with S. boulardii reduced the mean percentage of days with diarrhea per feeding days from 18.9 to 14.2% [odds ratio (OR) = 0.67, 95% confidence interval (CI) = 0.50-0.90, P = 0.0069]. In the control group, nine risk factors were significantly associated with diarrhea: nonsterile administration of nutrients in open containers, previous suspension of oral feeding, malnutrition, hypoalbuminemia, sepsis syndrome, multiple organ failure, presence of an infection site, fever or hypothermia, and use of antibiotics. Five independent factors were associated with diarrhea in a multivariate analysis: fever or hypothermia, malnutrition, hypoalbuminemia, previous suspension of oral feeding, and presence of an infection site. After adjustment for these factors, the preventive effect of S. boulardii on diarrhea was even more significant (OR = 0.61, 95% CI = 0.44-0.84, P < 0.0023). CONCLUSIONS: S. boulardii prevents diarrhea in critically ill tube-fed patients, especially in patients with risk factors for diarrhea. PMID- 9201524 TI - Validation of air tonometric measurement of gastric regional concentrations of CO2 in critically ill septic patients. AB - OBJECTIVE: To evaluate the accuracy of continuous air tonometry (Tonocap, Tonometric Division, Instrumentarium, Helsinki, Finland). DESIGN: The accuracy of air tonometry was tested by comparing it with conventional saline tonometry in mechanically ventilated, critically ill septic patients and in vitro determining the partial pressure of carbon dioxide (PCO2) of humidified gases with known concentrations of CO2. SETTING: A mixed intensive care unit in a university hospital. PATIENTS: 16 mechanically ventilated patients with sepsis. MEASUREMENTS AND RESULTS: Two gastric tonometer catheters (TRIP NGS catheter, Tonometric Division, Instrumentarium, Helsinki, Finland) were introduced into the patients' stomachs. The control catheter was used as a conventional saline tonometer and the other catheter was used with the Tonocap monitoring device. A total of 153 paired measurements was made and analysed according to Bland and Altman. The mean difference between air PCO2 and saline PCO2 values (bias), the standard deviation of the differences (precision), and the Pearson correlation coefficient between air PCO2 and saline PCO2 were calculated. The data on patients were pooled and calculated for different cycle times. The mean bias (kPa) was-0.02 with a 10-min cycle time, 0.31 with 15 min, 0.56 with 30 min and 0.21 with 60-min. The precisions were 0.39, 0.54, 0.44 and 0.76, respectively. Pearson correlation coefficients were 0.93, 0.97, 0.95 and 0.82, respectively (p < 0.0001). In vitro tonometry with the Tonocap was performed in a gas chamber fully saturated with known CO2 concentrations. The clinically important 10-min cycle time was tested with 5 Tonocap monitors. Except for the first 10-min cycle time, PCO2 values determined by the Tonocap monitoring systems were comparable to known CO2 concentrations. CONCLUSIONS: The accuracy of Tonocap continuous air tonometry is close to that of conventional saline tonometry. Moreover, the clinically important 10-min cycle time with air tonometry correlated very well with saline tonometry and the time response with air tonometry was short. PMID- 9201525 TI - Acute respiratory distress syndrome in a community hospital ICU. AB - OBJECTIVE: To estimate the incidence of the acute respiratory distress syndrome (ARDS) in an Australian urban community, and to describe the pattern of disease and outcomes in a community hospital intensive care unit (ICU). SETTING: An eight bed general ICU in a community hospital. DESIGN: Retrospective chart review. PATIENTS: 32 patients identified over a 4-year period as having ARDS. MEASUREMENTS AND RESULTS: The incidence of ARDS in an Australian urban community was estimated to be 7.3-9.3 cases/100,000 population per year. In-hospital mortality was 59%, while ICU mortality was 47%. Sepsis, pneumonia and aspiration were the main aetiological factors accounting for 94% of the patient population. There was no trauma. The Acute Physiology and Chronic Health Evaluation and Murray scores and values for the ratio of the partial pressure of oxygen in arterial blood and fractional inspired oxygen on admission to the ICU were similar between survivors and nonsurvivors, and none of these parameters were reliable predictors of outcome. Mean age, however, was different between survivors (56 +/- 16 years) and non-survivors (69 +/- 9 years) (p < or = 0.01). Mean daily fluid balance was also different between survivors (536 +/- 545 ml/day) and non-survivors (1576 +/- 1255 ml/day) (p < or = 0.02). Haemodynamic data were collected on 21 of the 32 patients within 72 h of the onset of ARDS. None of the haemodynamic parameters reached significance. There was, however, a trend for better cardiac function and oxygen consumption in the survivors. CONCLUSIONS: These data show that for ARDS, at least, mortality outcome can be comparable in a community ICU to a tertiary referral institution. The pattern of disease in an urban Australian community hospital is different to that often reported from tertiary centres. The incidence of ARDS in an Australian urban community is comparable to the reported incidence in North America and Western Europe. PMID- 9201526 TI - Respiratory system mechanics in the early phase of acute respiratory failure due to severe kyphoscoliosis. AB - OBJECTIVE: To evaluate respiratory mechanics in the early phase of decompensation in a group of seven patients with severe kyphoscoliosis (KS) (Cobb angle > 90 degrees) requiring mechanical ventilatory support. DESIGN: Prospective clinical study with a control group. SETTING: General intensive care unit at University of Rome "La Sapienza". PATIENTS: Seven consecutive patients affected by severe KS in the early phase of acute decompensation and a control group of six ASA (American Society of Anesthesiology) 1 subjects who were mechanically ventilated during minor surgery. MEASUREMENTS AND RESULTS: Respiratory mechanics were evaluated during constant flow-controlled mechanical ventilation at zero end-expiratory pressure with the end-inspiratory and end-expiratory occlusion technique. In five patients who showed increased ohmic resistance (RRSmin), we evaluated the possibility of reversing this increase with a charge dose of 6 mg/kg doxophylline i.v. In four KS patients, in whom a reliable esophageal pressure was confirmed by a positive occlusion test, we separated respiratory system data into lung and chest wall component. All KS patients showed reduced values of respiratory compliance (CRS) and increased respiratory resistance (RRS). The average basal values of CRS were 36 +/- 10 vs 58 +/- 8.5 cmH2O in control patients; RRSmax was 20 +/- 3.1 vs. 4.5 +/- 1.2 cmH2O/1 per s; RRSmin 6.2 +/- 1.2 vs. 2 +/- 0.5 cmH2O/1 per s: delta RRS 14 +/- 2.6 cmH2O vs 2.4 +/- 0.7 cmH2O/1 per s. All KS patients showed low values of intrinsic positive end-expiratory pressure (PEEPi) (1.8 +/- 1.5 cmH2O). Separation of lung and chest-wall mechanics, performed only in four patients, showed a reduction in both lung (66.7 +/- 7.2 ml/cmH2O) and chest wall values (84 +/- 8.2 ml/cmH2O), while both RmaxL and RmaxCW were increased (16.6 +/- 2 and 2.8 +/- 0.4 cmH2O/1 per s, respectively). Infusion of doxophylline did not significantly change respiratory mechanics when evaluated 15, 30, and 45 min after the infusion. CONCLUSIONS: During acute decompensation, both lung and chest-wall compliance are severely reduced in KS patients: conversely, and, contrary to that in patients with chronic obstructive pulmonary disease, increases in airway resistance and PEEPi seem to play only a secondary role. PMID- 9201527 TI - Breathing pattern and additional work of breathing in spontaneously breathing patients with different ventilatory demands during inspiratory pressure support and automatic tube compensation. AB - OBJECTIVE: We designed a new ventilatory mode to support spontaneously breathing, intubated patients and to improve weaning from mechanical ventilation. This mode, named Automatic Tube Compensation (ATC), compensates for the flow-dependent pressure drop across the endotracheal tube (ETT) and controls tracheal pressure to a constant value. In this study, we compared ATC with conventional patient triggered inspiratory pressure support (IPS). DESIGN: A prospective, interventional study. SETTING: A medical intensive care unit (ICU) and an ICU for heart and thoracic surgery in a university hospital. PATIENTS: We investigated two groups of intubated, spontaneously breathing patients: ten postoperative patients without lung injury, who had a normal minute ventilation (VE) of 7.6 +/- 1.7 l/min, and six critically ill patients who showed increased ventilatory demand (VE = 16.8 +/- 3.0 l/ min). INTERVENTIONS: We measured the breathing pattern [VE, tidal volume (VT), and respiratory rate (RR)] and additional work of breathing (WOBadd) due to ETT resistance and demand valve resistance. Measurements were performed under IPS of 5, 10, and 15 mbar and under ATC. RESULTS: The response of VT, RR, and WOBadd to different ventilatory modes was different in both patient groups, whereas VE remained unchanged. In postoperative patients, ATC, IPS of 10 mbar, and IPS of 15 mbar were sufficient to compensate for WOBadd. In contrast, WOBadd under IPS was greatly increased in patients with increased ventilatory demand, and only ATC was able to compensate for WOBadd. CONCLUSIONS: The breathing pattern response to IPS and ATC is different in patients with differing ventilatory demand. ATC, in contrast to IPS, is a suitable mode to compensate for WOBadd in patients with increased ventilatory demand. When WOBadd was avoided using ATC, the patients did not need additional pressure support. PMID- 9201528 TI - Left ventricular systolic and diastolic function in septic shock. AB - OBJECTIVE: The identification of myocardial dysfunction in septic shock has not yet been fully elucidated. We therefore studied patients with persistently vasopressor-dependent septic shock, both with invasive haemodynamic monitoring and transoesophageal two-dimensional and Doppler echocardiography (TEE). DESIGN: Prospective study. SETTING: General ICU in University Hospital. PATIENTS AND METHODS: All patients were monitored with arterial and pulmonary artery catheters. Haemodynamics were obtained concomitantly with TEE measurements. TEE was performed at three levels: a) a midpapillary short axis view of the left ventricle (LV) in order to measure end-systolic and end-diastolic areas; b) at the level of both the mitral valve for early (E) and late (A) filling parameters and c) the level of the right upper pulmonary vein for systolic (S) and diastolic (D) filling characteristics. Each parameter was characterised by maximal flow velocity and time velocity integral. RESULTS: Although the measurements of cardiac index demonstrated a wide range, three subsets of patients were identified post hoc after analysis on the basis of different Doppler patterns: first, patients with a LV without regional wall motion abnormalities and both E/A and S/D greater than 1 (group 1); second, patients with a comparable haemodynamic condition, apparently normal LV systolic function but with altered Doppler patterns: S/D less than 1 in conjunction with E/A more than 1 (group 2); finally, patients with compromised global LV systolic function, E/A less than 1 and S/D less than (group 3). CONCLUSIONS: Notwithstanding the known various interfering factors which limit the broad applicability of TEE to determine LV function in septic shock, our data suggest that cardiac dysfunction in septic shock shows a continuum from isolated diastolic dysfunction to both diastolic and systolic ventricular failure. These data strengthen the need of including the evaluation of pulmonary venous Doppler parameters in each investigation in order to obtain supplementary information to interpret diastolic function of the LV in septic shock patients. PMID- 9201529 TI - Nitric oxide synthase inhibition by L-NAME in leukocytopenic patients with severe septic shock. AB - OBJECTIVES: To investigate the effects of nitric oxide synthase inhibition by NG nitro-L-arginine methyl ester (L-NAME) on hemodynamics and outcome in leukocytopenic (< 1000/microliter) patients with severe septic shock requiring strong vasopressor support. DESIGN: Prospective clinical study. SETTING: Medical intensive care unit. PATIENTS: 10 patients with hematologic malignancies in chemotherapy-induced leukocytopenia with severe septic shock and high-dose vasopressor requirement. INTERVENTION: Continuous intravenous infusion of L-NAME (0.3 mg/ kg per hour) for a study period of 24 h with prolongation for up to 96 h according to individual requirements. MEASUREMENTS AND RESULTS: Compared to baseline values, an increase in mean arterial pressure (p = 0.0021), systemic vascular resistance (p = 0.0001), and left ventricular stroke work index (p = 0.023) with a concomitant decrease in vasopressor requirement (p < 0.05) was observed during the first 24 h of L-NAME treatment. Cardiac output data were unchanged during the study period (p = 0.49). L-NAME was tapered off in five patients who again became responsive to vasopressor medication. Two patients survived the episode of septic shock and vasoactive medication was stopped. CONCLUSIONS: The data demonstrate that inhibition of nitric oxide synthase may be beneficial for the treatment of severe septic shock in leukocytopenic patients as indicated by an increase in systemic vascular resistance, mean arterial pressure, and left ventricular stroke work index. PMID- 9201530 TI - Does continuous heparinization influence platelet function in the intensive care patient? AB - OBJECTIVE: To study the influence of continuous administration of heparin on platelet function in intensive care patients. DESIGN: Prospective, serial investigation. SETTING: Clinical investigation on a surgical and neurosurgical intensive care unit in a university hospital. PATIENTS: The study included 45 patients: 15 postoperative with patients sepsis (Acute Physiology and Chronic Health Evaluation II score between 15 and 25), 15 trauma patients (Injury Severity Score 15 to 25), and 15 neurosurgical patients. INTERVENTIONS: Management of the patients was carried out according to the guidelines for modern intensive care therapy. Sepsis and trauma patients received standard (unfractionated) heparin continuously [aim: an activated partial thromboplastin time (aPTT) approximately 2.0 times normal value; sepsis-heparin and trauma heparin patients], whereas neurosurgical patients received no heparin (neurosurgical patients). MEASUREMENTS AND RESULTS: From arterial blood samples, platelet aggregation was measured by the turbidimetric method. Platelet aggregation was induced by adenosine diphosphate (ADP; 2.0 mumol/l), collagen (10 micrograms/ml), and epinephrine (25 mumol/l). Measurements were carried out on the day of diagnosis of sepsis or 12 h after hemodynamic stabilization (trauma and neurosurgery patients) (baseline) and during the next 5 days at 12.00 noon. Standard coagulation parameters [platelet count and fibrinogen and antithrombin III (AT III) plasma concentrations] were also monitored. Heparin 4-10 U/kg per h (mean dose: approximately 500 U/h) was necessary to reach an aPTT of about 2.0 times normal. Platelet count was highest in the neurosurgical patients, but it did not decrease after heparin administration to the trauma and sepsis patients. AT III and fibrinogen plasma levels were similar in the three groups of patients. In the sepsis group, platelet aggregation variables decreased significantly (e.g., epinephrine-induced maximum platelet aggregation:-45 relative % from baseline value). Platelet function recovered during the study and even exceeded baseline values (e.g., ADP-induced maximum platelet aggregation: +42.5 relative % from baseline value). Continuous heparinization did not blunt this increase of platelet aggregation variables. In the heparinized trauma patients, platelet aggregation variables remained almost stable and were no different to platelet aggregation data in the untreated neurosurgical patients. CONCLUSIONS: Continuous administration of heparin with an average dose of approximately 500 U/h did not negatively influence platelet function in the trauma patients. Recovery from reduced platelet function in the sepsis group was not affected by continuous heparinization. Thus, continuous heparinization with this dose appears to be safe with regard to platelet function in the intensive care patient. PMID- 9201531 TI - Assessing the efficiency of the admission process to a critical care unit: does the literature allow the use of benchmarking? AB - OBJECTIVES: To determine the ability of the current literature to supply appropriate data for benchmarking admission practice to a multidisciplinary critical care unit. DESIGN: Retrospective review of data collected prospectively on a cohort of 614 patients and a systematic review of the literature. SETTING: A 30-bed multidisciplinary critical care unit at a university teaching hospital. PATIENTS: Consecutive admissions to the critical care unit over a 6-month period. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: For each patient, demographic data and admitting diagnosis were recorded on admission. Information necessary to calculate the Acute Physiology and Chronic Health Evaluation II and Therapeutic Intervention Scoring System (TISS) scores were collected daily. TISS variables were categorized as "active" or "non-active" treatment variables. Patients were then identified on a daily basis as receiving or not receiving active treatment. A review of the literature, using MEDLINE and the search term "Therapeutic Intervention Scoring Index" (as a textword), was conducted to identify studies that had similarly divided their patients. Using the method of benchmarking, the proportion of patients admitted who received active treatment during their stay in the critical care units was compared between the index critical care unit and those in the literature. A greater proportion of the patients admitted to our unit received active treatment (97.7%) when compared to other studies in the literature (20-66%). However, a number of potential confounding factors were present, such as the availability of intermediate care units, overnight recovery room ventilation, and critical care bed availability between the index critical care unit and those described in the literature. CONCLUSIONS: The current literature does not provide adequate data on critical care unit admission practices to allow useful application of the method of benchmarking. There is a need for publicly accessible large databases to allow individual critical care units to determine their level of efficiency when compared to similar institutions. PMID- 9201532 TI - Quality of life outcomes after intensive care. Comparison with a community group. AB - OBJECTIVE: Compare the health related quality of life of intensive care patients with a community sample. DESIGN: Self-completed questionnaire posted to a consecutive sample of 238 patients 16 months after discharge from an intensive care unit (ICU) and to a random community sample (n = 242). SETTING: The Liverpool Hospital is the main referral and teaching hospital in a community of 620,000 people. It has a ten-bed general ICU. PATIENTS AND PARTICIPANTS: All patients admitted to the ICU over 8 months with a length of stay > or = 24 h and a sample drawn from the community telephone directory. MEASUREMENTS AND MAIN RESULTS: The self completed questionnaire contained physical and psychosocial health and quality of life (QOL) scales. Analysis of variance indicated that ICU patients were more physically ill and anxiously depressed than the community sample. Sixty-three per cent of patients had not attained full health, were functionally impaired and had a poorer QOL than those patients who had returned to full health and the community. Psychosocial health (apart from anxious depression) was related to the level of perceived physical health rather than to whether or not they had been admitted to the ICU. Those subjects not in full health had poorer interpersonal relationships, less positive attitudes about life, more anxious depression and more suicidal depression. CONCLUSIONS: ICU patients following discharge have worse perceived health and more anxiety than others in the community. Sixty-three per cent of patients had a poorer QOL and functional health than those who returned to full health and those in the community. PMID- 9201533 TI - Detection of subclinical brain dysfunction by sensory evoked potentials in patients with severe diabetic ketoacidosis. AB - OBJECTIVE: Subclinical brain dysfunction is a potentially deleterious complication of diabetic ketoacidosis but is rarely recognized. Thus, we investigated the diagnostic value of sensory evoked potentials for detecting subclinical brain dysfunction in patients with diabetic ketoacidosis. DESIGN: Prospective trial. SETTING: Intensive care unit in a university hospital. PATIENTS: 5 neurologically asymptomatic patients (Glasgow Coma Scale score 15, slight drowsiness; aged 20 to 66 years) with an established diagnosis of severe diabetic ketoacidosis were studied. MEASUREMENTS AND RESULTS: Short- and long latency sensory evoked potentials were recorded within 2 h of initiation of therapy for ketoacidosis and 7 days after normalization of ketoacidosis, respectively. Two hours after starting therapy, sensory evoked potential peak latencies were prolonged in all five patients compared to age-matched healthy subjects [cervical N 13 to cortical N 20 interpeak latency of short-latency evoked potentials (mean) 5.8 vs 5.3 ms, p < 0.05; N 35 peak latency 40 vs 34 ms, p < 0.05; N 70 peak latency of long-latency evoked potentials 102 vs 76 ms, p < 0.01]. In all five patients, cervical N 13 to cortical N 20 interpeak latency and N 35 and N 70 peak latency reverted to normal 7 days after recovery from diabetic ketoacidosis. CONCLUSIONS: Our study indicates that the recording of sensory evoked potentials is a sensitive method of detecting subclinical brain dysfunction in patients with severe diabetic ketoacidosis. Since sensory evoked potentials were significantly prolonged in all five patients, this strongly suggests that subclinical brain dysfunction occurs more frequently than is generally recognized. PMID- 9201535 TI - Dysautonomy in daily activities impair the prognosis of acutely ill adults admitted to hospital. PMID- 9201534 TI - Neurological disturbances and hyperdynamic shock in a patient with esophagocoloplasty. AB - A 39-year-old man, with no history of alcohol intake, who had had an esophago ileo-colo-gastroplasty with ileotransversostomy, developed diplopia, seizures, metabolic acidosis, and cardiac failure and finally refractory hyperdynamic shock. He died 20 h after admission to our intensive care unit from cardiocirculatory collapse. Postmortem results revealed low erythrocyte transketolase activity, which was increased by 22% by in vitro addition of thiamine diphosphate (TDP effect). Cerebral pathology showed the alterations of Wernicke's encephalopathy. We discuss the possible mechanisms of fatal cardiovascular collapse and the unusual presentation of a case without a history of alcoholic intake or clinical malnutrition. PMID- 9201536 TI - Survival after cardiac arrest and severe acidosis (pH = 6.54) PMID- 9201537 TI - Severe arterial and venous thrombosis in a patient with heparin-induced thrombocytopenia type II. PMID- 9201538 TI - Severe hypokalemia causing rhabdomyolysis and quadriplegia in a patient with AIDS. PMID- 9201539 TI - Acute ammonia inhalation. PMID- 9201540 TI - Defibrillation in tricyclic antidepressant overdose. PMID- 9201541 TI - Fatal sodium valproate poisoning. PMID- 9201542 TI - Saline-lavaged lung injury and pressure support ventilation. PMID- 9201543 TI - Serious Clostridium difficile sepsis. PMID- 9201544 TI - Heterogeneity of corticotropin-releasing factor (CRF). AB - Although CRH is the major contributor to the physiological regulation of ACTH secretion, it must be recognized that a considerable degree of heterogeneity exists regarding the chemistry and biological activity of various CRF-active substances, some of which still remain to be fully clarified. The widespread extrahypothalamic sources and diverse extrapituitary actions of CRFs suggest that multiple CRF components must act in concert for the organism to cope with the environmental changes and stresses. PMID- 9201545 TI - The role of the pericellular fibrinolytic system in angiogenesis. PMID- 9201546 TI - Enhancement of splenic interferon-gamma, interleukin-2, and NK cytotoxicity by S36 acupoint acupuncture in F344 rats. AB - The effect of Tsusanli acupuncture point (S36 acupoint) stimulation on splenic natural killer (NK) cytotoxicity was examined in Fischer 344 (F344) rats. Electro acupuncture stimulation (voltage intensity, 1 to 5 V; duration, 1 ms; frequency, 1 Hz) was applied to bilateral S36 acupoints once a day (1 h) for 3 d. NK cytotoxicity was measured by the standard 4-h 51Cr release assay. Successive acupuncture treatment for 3 d significantly enhanced splenic NK cytotoxicity (p < 0.001) on the first day after final treatment as compared to that of the control. However, similar stimulation to abdominal muscle did not influence splenic NK cytotoxicity. We also examined endogenous cytokine activities in aqueous spleen extracts prepared from acupunctured and control rats. The extracts from rats acupunctured at the S36 acupoint contained high levels of interleukin (IL)-2 and interferon (IFN)-gamma as compared to those of abdominal muscle acupunctured and non-acupunctured control rats (p < 0.01). Furthermore, a significant positive correlation (p < 0.01) was observed between the levels of each cytokine tested and splenic NK cytotoxicity. The same positive correlation was also observed between the levels of IL-2 and IFN-gamma (p < 0.01). These observations indicate that electro-acupuncture stimulation of the S36 acupoint enhances splenic NK cytotoxicity and that IL-2 and IFN-gamma may function, at least in part, in the regulation of NK cell activity in this system. PMID- 9201547 TI - Optical mapping of conduction patterns of normal and tachycardia-like excitations in the rat atrium. AB - Multiple-site optical recording of transmembrane voltage activity (MSORTV), using a voltage-sensitive dye and a 12 x 12-element photodiode array, was employed to monitor action potentials in the rat atrium. Atrial preparations including the sinus node area, caval area, and atrial septum were dissected from adult rat hearts and stained with a voltage-sensitive merocyanine-rhodanine dye (NK2761). For suppression of optical artefacts due to contractile movements, a bathing medium containing 2,3-butanedione monoxime (BDM: 10-20 mM) was used. The spread of spontaneous excitation from the pacemaker was assessed optically by timing the initiation of the action potential-related extrinsic absorption changes. The optical signals were recorded from more than 300 contiguous loci in the atrium by sliding the photodiode array over the image of the preparation; in this way, the intra-atrial conduction pattern of spontaneous excitation was mapped. The obtained maps revealed a non-radial spread of excitatory waves originating in the pacemaking area over the atrium. Furthermore, we also mapped the conduction pattern of long-lasting tachycardia-like excitation evoked by a short train of electrical stimulation (5-10 Hz, 1-2s) applied with a bipolar electrode. These maps suggest that excitatory waves are propagated in a circular pathway which often surrounds the ostium of the superior or inferior vena cava. Various patterns for the pathway were also demonstrated optically. PMID- 9201548 TI - Permeation of organic cations and ammonium through the glutamate receptor channel in Drosophila larval muscle. AB - The inside-out configuration of the patch-clamp technique was used to study the effects of large organic cations on the single-channel current through the glutamate receptor channel in muscles of Drosophila larvae. Control experiments with symmetrical Na+ showed slightly supra-linear I-V curves. When external Na+ was equiosmolarly replaced with either arginine+ or N-methyl-D-glucamine+ (NMDG+), the reversal potential changed almost according to VNa, suggesting that both these ions are only slightly permeant through the channel. However, both ions strongly reduced the current amplitude in a voltage-dependent manner, the effect being most pronounced for the inward current. Tris+ had similar effects on the Na+ current, although the reversal potential indicated that this ion is somewhat permeant. All of these results could be fitted with a "one-ion" Eyring model for the channel with internal binding sites. The ion-channel dissociation constants for arginine+, NMDG+, and Tris+ were found to be 17, 27, and 23 mM, respectively. In order to acquire evidence for the "one-ion" hypothesis used in the model, I-V data were taken with different mixtures of Na+ and a comparably permeant ion, NH4+. All of the data could be accurately fitted with the results of the Eyring theory for singly occupied channels. PMID- 9201549 TI - Lipopolysaccharide-induced fever in rats prolonged by a needle prick. AB - We studied the effect of an abdominal prick with a needle on LPS-induced fever in freely-moving rats. LPS was injected intraperitoneally by the following 3 methods: 1) through a hypodermic needle pricked into the abdominal cavity, 2) through a catheter chronically indwelt in the abdominal cavity, and 3) through a catheter chronically indwelt in the abdominal cavity immediately after an abdominal prick was made. In the second method, core body temperature (Tb) began to rise about 1 h after the injection, reaching a maximal level at around 2.5 h and decreasing gradually thereafter. In the first and third methods, Tb rose again to make a second peak after making the first peak of fever. This was the same when LPS was injected through a hypodermic needle pricked into the abdominal cavity under restrained condition. These results suggest that the abdominal prick with a needle is responsible for the development of the second peak (or prolongation) of LPS-fever in rats. PMID- 9201551 TI - Photometric assessment of volume changes coupled with membrane potential in valinomycin-incorporated red blood cells. AB - Employing photometric methods, we have attempted to derive a possible quantitative relationship between volume change and membrane potential for valinomycin-incorporated red blood cells. The cells, collected from a human, rats or bullfrogs, were suspended in test solutions composed of a mixture of NaCl and KCl in varying proportions. The osmolality of the suspending medium was appropriately fixed at different values. After the addition of valinomycin to the suspension, changes in optical density (turbidity) at 620 nm and in fluorescence from a voltage-sensitive permeant dye (diS-C3-[5]) were measured in different concentrations of external potassium ions. The changes in optical density and fluorescence intensity were converted into relative cell volume and membrane potential changes in the test cells, respectively. Cell volume increased with depolarization of the membrane. We derived an empirical equation for the volume change versus membrane potential relation curves obtained experimentally, and have also shown that the observed volume change may be plausibly represented by a hyperbolic function of transmembrane potential involving the medium osmolality as an important parameter. PMID- 9201550 TI - 2,3-Butanedione monoxime suppresses excitation-contraction coupling in the canine blood-perfused left ventricle. AB - The negative inotropism of 2,3-butanedione monoxime (BDM) < or = 5 mmol/l has been attributed primarily to directly suppressed crossbridge force development without much suppressed intracellular Ca2+ handling. However, there is evidence that BDM simultaneously or even primarily suppresses myocardial excitation contraction (E-C) coupling. We therefore studied the mechanoenergetic effects of intracoronary BDM in the left ventricle (LV) of 11 canine excised cross circulated hearts. We fully utilized the VO2-PVA-Emax framework that we have developed, where VO2 is myocardial O2 consumption, PVA is the systolic pressure volume area as a measure of the total mechanical energy, and Emax is a contractility index. We gradually depressed Emax from 5.9 to 3.4 mmHg/(ml/100 g) on average by increasing intracoronary BDM to 2.6 +/- 2.1 mmol/l, and then gradually restored Emax to the pre-BDM level by increasing intracoronary CaCl2. We compared the O2 cost of Emax between BDM and Ca2+. We found that BDM and Ca2+ had a similar O2 cost of Emax. BDM did not affect the concentrations of blood borne catecholamines. We therefore conclude that the negative inotropism of BDM is primarily due to suppressed E-C coupling in canine blood-perfused hearts. PMID- 9201552 TI - Intracisternal injection of basic fibroblast growth factor reduces the severity of gastric mucosal lesions evoked by ethanol in rats. AB - This study was conducted to examine the hypothesis that basic fibroblast growth factor (bFGF) may have an anti-ulcer action through an acid-independent mechanism. The intracisternal injection of bFGF (1 microgram/10 microliters) significantly attenuated the development of gastric mucosal damage evoked by either subcutaneous indomethacin or intragastric absolute ethanol. On the other hand, intraperitoneally injected bFGF (1 microgram) failed to inhibit the formation of gastric mucosal injury by indomethacin or ethanol. These results suggest that bFGF acts in the brain to exert a gastroprotective action. Since ethanol-induced gastric lesion formation does not depend upon luminal acid, we speculate that an acid-independent mechanism may mediate the anti-ulcer action of central bFGF. PMID- 9201553 TI - ATP, thapsigargin and cAMP increase Ca2+ in rat hepatocytes by activating three different Ca2+ influx pathways. AB - The intracellular Ca2+ concentration ([Ca2+]i) in single isolated rat hepatocytes was measured using fura-2. Extracellular ATP induced La(3+)-sensitive and verapamil-insensitive Ca2+ influx together with Ca2+ release from intracellular Ca2+ stores. Incubation of hepatocytes with 2 microM thapsigargin produced a large prolonged increase in [Ca2+]i, which was insensitive to both 100 microM La3+ and 40 microM verapamil. Incubation with 1 mM dibutyryl cAMP increased [Ca2+]i in the presence of extracellular Ca2+ but did not in the absence of extracellular Ca2+, indicating that dibutyryl cAMP induces Ca2+ influx which was found to be sensitive to both La3+ and verapamil, without mobilizing Ca2+ from the intracellular pools. This study shows the presence of at least 3 different Ca2+ influx pathways in the plasma membrane: that is, 1) the ATP-induced, La(3+) sensitive and verapamil-insensitive pathway; 2) the thapsigargin-induced, La(3+) insensitive and verapamil-insensitive pathway; and 3) the cAMP-induced, La(3+) sensitive and verapamil-sensitive pathway. PMID- 9201554 TI - Effects of the angiotensin AT1 receptor antagonist GRI38950 on haemodynamic function in dogs. AB - 1. The antagonist activity of the angiotensin AT1 receptor antagonist, GR138950, and its haemodynamic effects have been investigated in beagle dogs. 2. In four anaesthetized dogs, GR138950 (0.1, 1 and 10 mg kg-1 i.v.), displaced dose response curves to angiotensin II (AngII) rightwards, in a parallel manner; GR138950, at 1 mg kg-1 i.v., produced a 15-fold displacement of the AngII dose response curve. In four conscious dogs, GR138950 (1 mg kg-1 i.v. or p.o.) antagonized AngII, and produced rightward and parallel displacements of the AngII dose-pressor response curves. Maximum displacements of approximately 33- and 16 fold occurred at 1 h after intravenous and 5 h after oral administration, respectively. The inhibitory effect of GR138950 was long lasting; AngII dose pressor response curves were still displaced by 10-fold from control values, 24 h after intravenous or oral administration of GR138950. 3. In comparison with its vehicle, GR138950 (0.1-10 mg kg-1) caused a significant decrease in resting blood pressure as a consequence of a significant decrease in total peripheral resistance in anaesthetized dogs. Effects on cardiac output, stroke volume and heart rate were not significantly different between GR138950- or vehicle-treated dogs. 4. Compared with vehicle, GR138950 administration did not significantly affect blood flow to the mesenteric and femoral vascular beds but did significantly increase blood flow to the renal vascular bed. Vascular resistance of the femoral bed was unaffected but that of the mesenteric and renal vascular beds was significantly reduced by GR138950, compared with the changes produced by vehicle treatment. 5. Compared with vehicle, left ventricular end diastolic pressure was significantly reduced by GR138950, but GR138950 had no significant effect on indices of cardiac contractility (+dP/dtmax and +dP/dt@40) or cardiac relaxation during early diastole (-dP/dt). 6. In conclusion, GR138950 acts as a competitive, surmountable antagonist at vascular angiotensin II receptors in beagle dogs. GR138950 reduced arterial blood pressure by reducing total peripheral resistance, attributable partly to reduction in resistance in the mesenteric and renal vascular beds. GR138950 reduced left ventricular end diastolic pressure whilst having no direct effect on cardiac contractility or relaxation. PMID- 9201555 TI - Inhibition of vasodilator neurotransmission in the sheep middle cerebral artery by VIP antiserum. AB - 1. Middle cerebral artery rings from the sheep were relaxed by vasoactive intestinal peptide (VIP) (20-200 nM) or by stimulation of the vasodilator nerves, electrical field stimulation (EFS). 2. VIP antiserum 1:256 inhibited the relaxation produced by both exogenous VIP (from 70 +/- 5 to 32 +/- 9% of 5-HT induced tone at 200 nM VIP) and by EFS (from 43 +/- 5 to 26 +/- 6% of 5-HT induced tone after 20 min), while pre-immune serum was inactive. 3. Capsaicin (1 microM) produced a transient relaxation but did not alter the response to EFS which was subsequently inhibited by addition of L-NOArg (100 microM). VIP-induced relaxation was antagonized by L-NOArg (50 microM) (from 68 +/- 5 to 46 +/- 2% relaxation of 5-HT-induced tone) but not by D-NOArg. 4. Exogenous VIP produced an approximately 2.4-fold increase in cyclic GMP content which was prevented by preincubation with L-NOArg (100 microM) but not D-NOArg. 5. VIP and neuronal NOS immunoreactivity was co-localized to the same adventitial nerve fibres in the sheep middle cerebral artery. 6. These results provide evidence that neurogenic relaxation in the sheep middle cerebral artery is, at least in part, mediated by VIP, involving activation of NO synthase. PMID- 9201556 TI - Pharmacological characterization of the muscarinic receptor subtypes responsible for the contractile response in the rat urinary bladder. AB - 1. Contractile responses of smooth muscle from the Wistar rat urinary bladder were studied with the use of muscarinic agonists and antagonists. 2. McN-A-343 induced only weak contractile responses of the bladder muscle. In contrast, oxotremorine showed higher potency than either acetylcholine or bethanechol in inducing a contractile response (the respective pD2 values were 6.38 +/- 0.25, 4.82 +/- 0.24 and 4.42 +/- 0.14). 3. The M2 antagonists, methoctramine (10(-9) M to 10(-5) M) and gallamine (10(-9) M to 10(-5) M), did not reduce acetylcholine induced (10(-5) M) contractions of the bladder muscle strip. On the other hand, 4 diphenyl-acetoxy-N-methyl piperidine methiodide (4-DAMP, 10(-10) M to 10(-7) M), an M3 receptor blocker, effectively antagonized the acetylcholine-induced contractions in a concentration-dependent manner. 4-DAMP had a similar pA2 value to those of the non-selective antagonists, atropine and scopolamine (pA2 values were 8.26 +/- 0.05, 8.36 +/- 0.05 and 8.41 +/- 0.11, respectively). Pirenzepine, and M1 blocker, antagonized the contractions at higher concentrations (10(-8) M to 10(-5) M, pA2 = 6.23 +/- 0.04). 4. It is concluded that (1) the dominant muscarinic receptor subtype responsible for smooth muscle contraction in the rat urinary bladder is M3; and (2) the muscarinic agonist oxotremorine was more potent than acetylcholine and bethanechol in inducing a contractile response. PMID- 9201557 TI - Spectral analysis of intercycle heart fluctuations in the diethyl-ether anaesthetized or pithed rat treated with l-hyoscyamine. AB - 1. Within the context of neural regulation of the activity of sinus node pacemaker cells, the study of heart rate variability, as explored in the frequency domain by spectral analysis, was proposed about 15 years ago as a quantitative tool for the evaluation of short-term autonomic cardiovascular control. It has since been postulated that the two main oscillations observed, one at low and the other at high frequency, may respectively be markers of sympathetic vs. vagal efferent cardiac activity, and that the low- and high frequency signals may reflect a reciprocal or 'push-pull' relationship between sympathetic and parasympathetic control. 2. In our power spectra assessment, ECG R-R intervals were submitted to fast Fourier transformation analysis in order to study the mechanisms underlying the control of heart beats in rats. Data were acquired in conditions of steady arterial blood pressure and cardiac and respiratory activity (spontaneous or artificially stimulated) in diethyl-ether anaesthetized and pithed rats, as well as in a group of control rats, all in the presence and absence of l-hyoscyamine. 3. With increasing doses of the parasympathetic antagonist, the fractal dimension of the time-series structure remained stable in most cases. The low-frequency spectral component narrowed with increasing drug doses and the high-frequency band underwent either no, or only very slight, changes. 4. In these rodent assays, the low- and high-frequency signals cannot be interpreted as a push-pull relationship between sympathetic and parasympathetic control. PMID- 9201558 TI - Suppression of the rat micturition reflex by imipramine. AB - 1. This study investigates possible mechanisms through which imipramine (IMI) exerts its antienuretic effect. The micturition reflex in response to bladder distension produced by saline infusion was examined in anaesthetized rats. 2. The amplitude and frequency of micturition reflex contractions were reduced by peripheral administration of IMI, but the micturition reflex was abolished after its intracerebroventricular (i.c.v.) administration. A muscarinic antagonist, atropine, displayed an inhibitory effect similar to that of IMI. A muscarinic agonist, carbachol, produced a dose-related rightward shift of the dose-response curve to IMI. Both IMI i.c.v. and the muscarinic antagonist l-methylscopolamine i.c.v. elevated the threshold of volume and pressure for micturition initiation, indicating that IMI and muscarinic antagonists mainly exert a central inhibitory effect on the micturition reflex. 3. In addition, to evaluate the role of central monoaminergic neurotransmission on micturition, the acetylcholine depletor hemicholinium-3 (HC-3), the catecholamines depletor alpha-methyl-p-tyrosine (AMPT), and the serotonin depletor p-chlorophenylalanine (PCPA) were examined alone or in combination with IMI. The micturition threshold was increased by treatment with HC-3, but not by AMPT or PCPA. In HC-3 treated rats, the inhibitory effect of IMI on the micturition reflex was more prolonged than in normal rats. After administration of IMI, the recovery from the cessation of micturition reflex contractions was facilitated by carbachol in normal rats, but not in HC-3 treated rats. This indicates that acetylcholine plays a facilitatory role in initiating micturition reflex contractions. 4. Acute treatment with IMI decreased the frequency and increased the volume threshold of micturition reflex contraction. Acute and chronic treatment with IMI prolonged the cessation period of micturition by IMI. 5. These results suggest that IMI exerts an inhibitory action on the micturition reflex by a central cholinergic mechanism. Muscarinic receptors located at the supraspinal level are tonically stimulated during distension-induced micturition reflex. PMID- 9201559 TI - Pharmacological modulation of electromechanical coupling in the proximal and distal regions of the guinea-pig renal pelvis. AB - 1. The effect of drugs affecting calcium and potassium channels and intracellular calcium handling/release on electromechanical coupling in the smooth muscle of the guinea-pig proximal vs. distal renal pelvis were investigated by using the single sucrose gap method. 2. Spontaneous action potentials discharged from the proximal renal pelvis were bell-shaped, did not show a pronounced plateau and had a small after-hyperpolarization. Spontaneous action potentials from the distal renal pelvis were characterized by a fast depolarization, a pronounced plateau and after-hyperpolarization. 3. Nifedipine (1 microM) suppressed action potentials in both regions of the renal pelvis. A submaximally effective concentration of nifedipine (50 nM) shortened action potential duration and reduced contractility in both regions of the renal pelvis. On the other hand Bay K 8644 (1 microM) markedly prolonged the duration of the action potential and increased contractility in both regions of the renal pelvis. 4. Tetraethylammonium (0.5 mM) markedly prolonged the action potential duration and contraction in the distal renal pelvis without affecting action potentials in the proximal renal pelvis. Similar effects were produced by a slightly higher concentration of tetraethylammonium (2 mM) in the proximal renal pelvis. 5. Charybdotoxin (30 nM) markedly prolonged the duration of action potential and increased and prolonged the contraction in both the proximal and distal renal pelvis. 6. 4-aminopyridine (1 mM) selectively increased the frequency of action potentials in the distal renal pelvis without affecting other parameters of the action potential nor contractility. 4-aminopyridine had no effect in the proximal renal pelvis. 7. The inhibitor of sarcoplasmic reticulum Ca-ATPase, cyclopiazonic acid (10 microM) transiently increased the frequency of action potentials in both regions of the renal pelvis; CPA markedly delayed the repolarizing phase of the action potential in both the proximal and distal renal pelvis and, in parallel, increased contractility. 8. We conclude that action potentials generated from the proximal and distal regions of the guinea-pig renal pelvis are evenly dependent upon the availability of L-type Ca channels; that Ca-dependent maxi K channels provide a major contribution to the repolarization of action potentials in both regions of the renal pelvis, thus regulating duration/intensity of Ca influx and contraction; that release of Ca from the internal store is not important in providing activator Ca for contraction but regulates duration of the action potential and may be involved in setting the frequency of discharge of pacemaker cells. PMID- 9201561 TI - On needles, breasts and bullets: health and the conflict of values. PMID- 9201560 TI - Effect of 4-aminopyridine and 3,4-diaminopyridine on guinea-pig isolated ileum. AB - 1. 4-Aminopyridine and 3,4-diaminopyridine produced concentration-dependent contraction on guinea-pig isolated ileum incubated in Tyrode solution. The EC30 values were 1.14 x 10(-4) and 1.39 x 10(-4) M, respectively. 2. Calcium channel blockers such as verapamil, diltiazem, nifedipine, flunarizine, and lanthanum chloride antagonized the contracting effect induced by 4-aminopyridine and 3,4 diaminopyridine in guinea-pig isolated ileum. 3. Diazoxide and atropine sulphate behaved similarly as antagonists of the contracting effect induced by 4 aminopyridine and 3,4-diaminopyridine in guinea-pig isolated ileum. 4. It is concluded that the aminopyridines exert their effects through the release of acetylcholine from parasympathetic nerve terminals. PMID- 9201562 TI - Growth hormone treatment for growth hormone deficient adults. AB - Growth hormone (GH) deficiency in adults is now recognized as a clinical syndrome with characteristic signs and symptoms. Numerous trials with daily subcutaneous biosynthetic human growth hormone (hGH) have been conducted in this patient group. Generally, improvements in insulin-like growth factor levels, decreases in total fat mass and increases in lean body mass are recorded with no overall effect on total body weight. Variable effects on serum cholesterol, bone mineral density and quality of life have also been reported. The true place of GH replacement therapy in adults has yet to be defined. Several questions relating to the dose, duration of treatment, long-term side-effects, quality of life changes and health economic implications of treatment still need to be assessed. PMID- 9201563 TI - Therapeutic advances: riluzole for the treatment of motor neurone disease. AB - Amyotrophic laterial sclerosis is a fatal neurogenerative disorder, for which only symptomatic treatment was previously available. Riluzole was recently launched in the U.S.A. and Europe. This is the first drug to produce a modest increase in survival (approximately 3 months) in patients with this disease. Unfortunately, treatment causes a number of side-effects of which asthenia is particularly troublesome. Between 10 and 20 per cent of patients can be expected to withdraw from treatment due to these adverse effects. Data on the real time survival advantage and quality of life that can be expected whilst on treatment are needed to identify the place of riluzole in the management of this distressing disease. PMID- 9201565 TI - Plasma concentrations of vigabatrin in epileptic patients. AB - OBJECTIVE: To measure plasma concentrations of vigabatrin in a group of 30 adult epileptic patients with complex partial seizures (CPS) refractory to conventional antiepileptic drugs. With a view to better defining the drug's dose-response relationships in the presence of concomitant alternative antiepileptic drugs. METHODS: High-performance liquid chromatographic analysis of blood samples drawn at steady-state trough levels from patients receiving vigabatrin. RESULTS: The steady-state plasma concentrations of vigabatrin showed marked interpatient variability (CV = 59.5%). The mean concentration was 42 +/- 25 micrograms/ml (range 11.5-102.7 micrograms/ml). Nineteen patients (63%) had plasma levels between 20 and 60 micrograms/ml. The plasma clearance of vigabatrin ranged from 0.24 to 1.37 ml/min/kg (mean +/- SD = 0.74 +/- 0.40 ml/min/kg), with a median value of 0.64 ml/min/kg. Concomitant treatment with carbamazepine increased the plasma clearance of vigabatrin from 0.59 ml/min/kg (monotherapy), 0.54 ml/min/kg (co-treated with phenobarbital) and 0.41 ml/min/kg (co-treated with valproic acid) to 0.92 ml/min/kg. CONCLUSION: Vigabatrin plasma levels show wide interpatient variability, and co-administration with carbamazepine increases vigabatrin's clearance. While there is no relationship between plasma level and anti-epileptic effect, abnormally high levels of the drug may increase toxicity. PMID- 9201564 TI - The rationale for new therapies in acute ischaemic stroke. AB - Although stroke is a major cause of morbidity and mortality, it is only relatively recently that a concerted effort has been made to develop acute treatments. Thrombolytics, such as recombinant tissue plasminogen activator (rt PA), may benefit selected patients within 3 h of cerebral infarction. CUrrently, rt-PA is only licensed for use in the United States. Many potential strategies for neuroprotection exist and are currently under investigation. Because the mechanisms of neurotoxicity involve numerous interdependent processes, it may be that the interpretation of a single site in the cascade of events is insufficient to provide effective neuroprotection. Drugs acting at several sites in the neurotoxic cascade may be more effective, and the results of Phase III studies with the novel neoroprotectant lubeluzole are anticipated. PMID- 9201567 TI - Does once-daily dosing of aminoglycosides affect neuromuscular function? AB - OBJECTIVE: Aminoglycosides have been reported to produce a curare-like neuromuscular blockade in animals at serum concentrations higher than those obtained with traditional dosing (1-2 mg/kg every 8 h) in humans. Aminoglycoside induced neuromuscular blockade is rarely, if ever, seen in humans with traditional dosing. The recent adoption of once-daily dosing of aminoglycosides has raised concerns about increased potential for this adverse effect because higher serum concentrations are produced. The objective of this study was to determine if once-daily dosing of aminoglycosides inhibits respiratory muscle function. METHOD: Nine mechanically ventilated ICU patients on once-daily dosing of gentamicin 6 mg/kg/day were assessed for respiratory muscle strength by measuring maximum inspiratory pressure (MIP). MIP is a measurement of the maximal negative pressure generated by repeated inhalations against an occluded airway over 20 s. This was measured within 1 hour before (MIPpre) and within 1 hour after each aminoglycoside dose (MIPpost). RESULTS: Mean values for MIPpre and MIPpost were -26.7 cm H2O and -26.5 cm H2O, respectively. The mean difference between MIPpre and MIPpost was -0.2 cm H2O, which was not statistically significant (P > 0.05). CONCLUSION: The effect of gentamicin (6 mg/kg/day) on respiratory muscle function was not statistically, nor clinically significant, and weaning from mechanical ventilation does not seem to be inhibited by once daily dosing of aminoglycosides as detectable by measurement of MIP. PMID- 9201566 TI - Educating the public about skin cancer prevention: a role for pharmacists. AB - OBJECTIVES: The incidence of skin cancer is increasing at an alarming rate. This study aimed to assess pharmacists' willingness to counsel patients about skin cancer and factors which are predictive of this. METHODS: A 30 item survey assessing skin cancer prevention counselling attitudes and behaviours, personal skin cancer prevention behaviours, and willingness to counsel on skin cancer prevention, was mailed to 300 randomly selected San Diego County pharmacists. RESULTS: Data obtained from 128 pharmacists (43% response rate) indicated that the incidence of skin cancer prevention counselling was very low, with over one third stating that they never counsel patients on this topic. Of those that did counsel, the percentage of patients counselled on skin cancer prevention was below 5%. Bivariate analyses indicated that pharmacists who counselled on this topic differed from those that did not on knowledge of skin cancer, attitudes and beliefs about counselling patients, pharmacy setting, services offered by their pharmacies, and their personal sun protective behaviours. Multivariate analysis indicated that two variables independently predicted skin cancer prevention counselling; whether the pharmacy where the respondent was employed offered this service, and attitude about counselling. Over 90% of respondents reported they were willing to counsel patients on skin cancer prevention and other health related topics. CONCLUSIONS: Future studies should focus on skin cancer prevention counselling barriers and environmental prompts encouraging this type of interaction between pharmacists and patients. PMID- 9201568 TI - An audit of discharge medication for secondary prophylaxis post-myocardial infarction. AB - BACKGROUND AND OBJECTIVE: Prophylactic pharmacotherapy post-myocardial infarction (post-MI) has been shown to reduce the risk of subsequent non-fatal infarction and to reduce overall mortality. The present study aimed to review the therapy of such patients in a teaching hospital. METHOD: Medicines prescribed to 77 post-MI patients for secondary prophylaxis were reviewed. The drugs prescribed by a cardiologist-led team were compared with those prescribed by a general medical team. Seventeen patients died before discharge, leaving a study cohort of 60 patients to complete the audit. RESULTS: Thirty-nine of the 60 patients (65%) met the discharge standard, and there was no significant difference in adherence to the audit standard when the consultant cardiologist and the general medical consultant were compared. The audit highlighted the therapeutic dilemma concerning the appropriateness of initiating low-dose aspirin in patients already receiving warfarin therapy. CONCLUSION: The audit has enabled us to develop guidelines which feed into the multi-disciplinary cardiac rehabilitation programme, helping to improve prescribing adherence to the discharge standard. A follow-up audit is planned. PMID- 9201569 TI - Pharmacokinetics of amikacin in intensive care unit patients. AB - OBJECTIVE: To characterize the population pharmacokinetics of amikacin in intensive care unit (ICU) patients and to analyse whether these patients show different kinetic behaviour on the basis of their clinical diagnoses. METHOD: The patient population comprised 104 medical ICU patients on amikacin treatment for several presumed or documented Gram-negative infections. Four study groups were defined according to patients' clinical diagnosis: sepsis group (n = 39), trauma group (n = 20), pneumonia group (n = 21) and 'other diagnosis' group (n = 24). The pharmacokinetic parameters for amikacin in these patients were then compared. RESULTS: The ICU patients were found to have increased values for the amikacin volume of distribution (0.52 +/- 0.21 litres/kg), whereas total amikacin clearance expressed as a linear function of creatinine clearance was Cl (ml/min/kg) = 0.13 +/- 0.86 ClCR, which is not significantly different from other estimations reported in the literature. However, this relationship revealed statistically significant differences among the four groups of ICU patients. Moreover, the septic and trauma patients showed higher (but not statistically significant) values for the amikacin volume of distribution. CONCLUSION: The amikacin pharmacokinetic parameters obtained should allow Bayesian individualization of amikacin doses in patients admitted to medical ICUs, on the basis of their clinical diagnoses. PMID- 9201570 TI - The influence of Rx-to-OTC changes on drug sales. Experiences from Sweden 1980 1994. AB - OBJECTIVE: A descriptive study of national sales data from Sweden to find out the effect of over-the-counter (OTC) switches on total sales of a number of drugs. During the period 1980-94, 16 drugs were changed from prescription only (Rx) status to the Swedish OTC market. Total sales increased for 14 out of these 16 drugs. The increase was seen soon after the change to OTC status. Two years after the change an average increase of 36% was seen. In the following 2 years, the increase was typically very modest (average 1%). Large differences in the changes were seen for the individual drugs. The prescription of OTC packs decreased on average by 26% during the first 2 years after the switch. Converting this decrease in sales in terms of number of packs no longer prescribed led to an estimated yearly saving of SEK 200 million ($US 30 million) for the national drug budget. Taking account of the total increase in defined daily doses (DDDs) 2 years after the change for those 16 drugs led to an estimated yearly saving of SEK 2.5 billion ($US 400 million). PMID- 9201571 TI - Survival of lactic acid bacteria in a dynamic model of the stomach and small intestine: validation and the effects of bile. AB - This study was conducted to validate a dynamic model of the stomach and small intestine to quantify the survival of lactic acid bacteria and to assess the influence of gastrointestinal secretions. The survival of a single strain of each of the following species, Bifidobacterium bifidum, Lactobacillus acidophilus, Lactobacillus bulgaricus, and Streptococcus thermophilus, was measured under physiological conditions (e.g., peristalsis, changes in pH, and changes in concentrations of enzymes and bile) and were compared with data obtained from humans. No significant differences were found between the in vitro and in vivo data, indicating that the model has a predictive value for the survival of these bacteria in humans. The survival of these strains of lactic acid bacteria in the gastrointestinal model was investigated under two different conditions in the small intestine: simulation of physiological secretion of bile and low bile secretion. Reductions in viability were significantly different between the bacterial species. The dose-response effect of bile on the survival of the tested bacteria was significant, demonstrating the bactericidal effect of bile salts. This study demonstrates the differences among bacterial species in their sensitivity to gastric and intestinal secretions. PMID- 9201572 TI - Influence of standards and antibodies in immunochemical assays for quantitation of immunoglobulin G in bovine milk. AB - This study reports the influence of using various reference antigens or standards as well as the source of antibody in the immunochemical quantitation of bovine immunoglobulin (Ig) G. Standard curves from analyses by ELISA and radial immunodiffusion were compared for bovine IgG from serum, colostrum, and cheese whey and for the two subclasses IgG1 and IgG2. Also compared were different sources of polyclonal antibodies (antisera from rabbit and sheep or hen yolk) and monoclonal antibodies that had various antibody specificities. The results indicate that IgG1 was a reliable alternative to purified IgG from milk or cheese whey for quantitation of IgG in milk based on ELISA absorbance. Serum IgG, colostral IgG, and IgG2 greatly underestimated milk IgG, regardless of the source of antibody used. Determination of milk IgG by radial immunodiffusion using antisera that were specific for rabbit anti-bovine IgG (H + L) was less dependent on the source of IgG that was used as the standard antigen. However, radial immunodiffusion using subclass-specific antibodies led to inaccurate estimation of IgG in milk unless the quantitation was based on a standard curve for IgG from milk. The Ig from hen yolk were a feasible alternative source of specific antibodies for immunoassay of IgG in bovine milk. PMID- 9201573 TI - Binding of retinoids to beta-lactoglobulin isolated by bioselective adsorption. AB - Binding of the retinoids, all-trans-retinol, all-trans-retinal, all-trans-retinyl acetate, and all-trans-retinoic acid, to beta-lactoglobulin (LG) (96% purity) that had been prepared by bioselective adsorption on N-retinyl-Celite was determined from changes in the fluorescence quenching (332 nm) of the protein tryptophanyl residues. High affinity binding of all of these compounds occurred at pH 7.0, and the apparent dissociation constant ranged from 1.7 to 3.6 x 10(-8) M. Furthermore, a stoichiometry of 1.0 mol.mol-1 of protein was obtained for each case, indicating that all of the sites in the protein preparation were available. When beta-LG in whey protein isolate (57.4% beta-LG) was studied, a stoichiometry of 0.65 to 0.82 mol.mol-1 of protein was obtained, indicating that a large number of the sites already had bound lipid or that the protein had been denatured. As the pH was lowered toward 5.15, the affinity decreased about fourfold, but the stoichiometry of binding was unchanged. Far UV circular dichroism spectra indicated that the secondary structure of the protein was not significantly affected by ligand binding; however, the near UV spectra were changed, indicating that the flexibility of tryptophanyl residues decreased. The latter effect is consistent with the change in fluorescence quenching and suggests that a tryptophan is in the binding site. PMID- 9201574 TI - Binding of vitamin D and cholesterol to beta-lactoglobulin. AB - beta-Lactoglobulin was isolated directly from acidic whey by bioselective adsorption on N-retinyl-Celite, yielding preparations of > or = 96% purity. Interactions of these preparations with vitamin D2, vitamin D3, ergosterol, cholesterol, and 7-dehydrocholesterol were examined by following changes in the fluorescence spectra. Both the excitation and emission spectra indicated that energy was transferred between the tryptophanyl residues of the protein and the chromophore of the ligand. Analyses of the fluorescence changes that occurred upon titration of beta-LG with the various ligands allowed determination of the dissociation constant for the complex and the number of moles bound per mole of protein. The affinity for vitamin D2 (dissociation constant of 4.91 nM) was 10 fold higher than that of the other compounds, except for ergosterol, which was 5 fold larger than the others. Also, the affinity was 10-fold higher than that typically reported for the retinoids. Furthermore, the value obtained for the number of moles bound per mole of protein was 2 mol.mol-1 for each of the ligands examined in this study; it has been well established that all of the retinoids are bound with a stoichiometry of 1.0. These results suggest that beta-LG may be a better carrier of vitamin D than of vitamin A. PMID- 9201575 TI - Empirical Bayes prediction of 305-day milk production. AB - A lactation curve described by an algebraic formula can be fitted by regression to the milk weights in the partial record of an individual lactation in progress; however, curves that are obtained in this manner do not provide useful predictions of milk production throughout the remainder of the lactation. This study examined the reasons for this failure and introduced a new empirical Bayes statistical method for fitting Wood's curve that was designed to provide good predictions of future production. The results of a comparison between predictions produced by the new method and predictions from Dairy Herd Improvement Association extension factors were quite favorable to the new method, which has advantages other than greater accuracy; the method does not require preparation of extension factor tables and can be readily adapted to individual herds. Comparisons revealed features of the Dairy Herd Improvement Association predictions that limit their usefulness for some herd management purposes. PMID- 9201576 TI - Role of insulin in the regulation of milk fat synthesis in dairy cows. AB - Five lactating Holstein cows were fitted with rumen fistulas and subjected to a hyperinsulinemic-euglycemic clamp and abomasal casein infusion to examine the effects on milk fat synthesis and the composition of milk fatty acids. The experiment consisted of two periods of abomasal infusions (water or 0.5 kg/d of casein); each period was divided into three 4-d intervals. The initial interval allowed for acclimation, and baseline measurements were established during the second interval. During the third 4-d interval, a hyperinsulinemic-euglycemic clamp was maintained, and insulin was infused continuously at the rate of 1 microgram/kg of body weight per h. Circulating concentrations of insulin were increased more than fourfold, and euglycemia was maintained by infusion of glucose at variable rates. Insulin had no effect on milk fat yield but casein infusion increased milk yield and tended to increase fat yield. A trend toward higher milk yield during the clamp, combined with a slight numerical decrease in milk fat yield, resulted in decreased fat percentage. Calculated net energy balance was positive throughout the study, although feed intake decreased during the insulin clamp, particularly for the water infusion period. Minor changes occurred in the composition of milk fatty acids during the clamp when the balance between de novo and preformed fatty acids shifted slightly toward de novo. Overall, results demonstrated that a relatively constant rate of milk fat synthesis was maintained during chronic hyperinsulinemia. Effects on milk fat yield and composition of fatty acids offered no support for the role of insulin on milk fat depression. PMID- 9201577 TI - The effect of bovine somatotropin and diet on somatotropin binding sites in hepatic tissue of lactating dairy cows. AB - In the lactating cow, galactopoiesis is stimulated by treatment with recombinant bovine somatotropin (bST) and by an improved plane of nutrition. The present study determined the interaction between these variables and examined whether a positive galactopoietic effect was accompanied by a change in hepatic binding sites for bST. Lactating dairy cows received one of three diets with increasing nutrient density; diet 1, 150 g/kg of dry matter (DM) of crude protein (CP) and 10.5 MJ/kg of DM of metabolizable energy; diet 2, 170 g/kg of DM of CP and 11.3 MJ/kg of DM of metabolizable energy; and diet 3, 190 g/kg of DM of CP and 12.1 MJ/kg of DM of metabolizable energy. At 90 d after calving, half of the cows in each dietary group were treated with bST every 14 d for the rest of the lactation. Both nutrient density and administration of bST increased milk yield significantly in mid and late lactation; there was no significant treatment by diet interaction. Treatment with bST significantly increased plasma concentrations of insulin-like growth factor (IGF)-I compared with IGF-I concentrations in controls in both mid and late lactation. Comparisons within diet revealed that concentrations of IGF-I were significantly higher in cows fed diet 3 than in cows fed diets 1 and 2 at both stages of lactation. Increases in plasma insulin were confined to cows in late lactation, and no changes were observed for nonesterified fatty acids. Liver biopsies showed that concentrations of hepatic binding sites for bST were not affected significantly by bST treatment but were increased in midlactation for cows fed diet 3. Concentration of hepatic binding sites per unit weight of tissue were greater for cows in midlactation than for cows in late lactation. In summary, exogenous bST treatment and increased nutrient density were associated with elevated plasma IGF-I concentrations and increased milk yield; however, only nutrient density in midlactation increased the number of hepatic binding sites for bST. Exogenous bST treatment had relatively little effect on the concentration of hepatic bST receptors compared with nutrient density. PMID- 9201578 TI - The empirical impact of bovine somatotropin on New York dairy farms. AB - Data from 259 farms that participated in the Cornell University Dairy Farm Business Summary Project were used to measure the impact of bovine somatotropin (bST) on milk production and profitability per cow. Linear regression was used to estimate the response to bST when other variables were held constant. Bovine somatotropin was a dummy variable for bST use or nonuse in the equations. For farms that used bST on 25% or more of the cow days in 1994, milk production increased by 510 kg per cow. Returns above purchased feed costs increased $153 per cow for farms that used bST. Milk receipts over operating costs increased $120 per cow for farms that used bST. The use of bST was estimated to increase net farm income by $27 per cow, but this estimated coefficient was not statistically different from 0. Differences in costs for labor or veterinary services per cow were not significantly different between farms that used bST and farms that did not use bST. PMID- 9201579 TI - Predicting optimal time of insemination in cows that show visual signs of estrus by estimating onset of estrus with pedometers. AB - An experiment was designed to estimate the optimal interval from the beginning of estrus to artificial insemination (AI). The data were analyzed by means of a mathematical model. The analysis was based on pedometer readings and results of rectal palpation at 42 to 49 d post-AI of 171 breedings in 121 cows. The chance of conception was highest between 6 and 17 h after increased pedometer activity; the estimated optimum was at 11.8 h. In this data file, the effects of disease, inseminator, time of AI (a.m. or p.m.), and bull did not contribute to the improvement of the model. The effects of disease were not significant because of the low incidence of any specific disease. Activity measurements can be used as a tool for AI strategy to improve conception in groups of healthy cows and heifers already showing visual signs of estrus. PMID- 9201580 TI - Discrimination of people by dairy cows based on handling. AB - This study examined whether dairy cows could distinguish among people based on the treatment received, whether cows used color as a cue to make this discrimination, and whether cows generalized their discrimination to other locations. Twelve cows were each repeatedly treated in a special treatment stall by two people wearing red or yellow overalls. One person always treated the cows aversively, and the other always treated them gently. The distance between each person and each cow in the home stall and in the treatment stall was scored during tests. Before treatment, the distances that cows maintained from the two people were uncorrelated, and the distances that they maintained in the treatment stall were uncorrelated with those in the home stall. Before and after treatments, the cows stood further from the handlers in the treatment stall than in the home stall, regardless of color of the overalls. Defecation and urination were more frequent during aversive treatments. After treatment, the cows stood further from the aversive handler than from the gentle handler in both stalls, and distance from the aversive handler was positively correlated with distance from the gentle handler. The cows did not discriminate when the aversive and gentle handlers wore blue overalls (as worn by the usual barn handlers), when two unfamiliar people wore the same color overalls as the handlers, or when the cows were shown photographic slides of the two handlers. In conclusion, the cows learned to discriminate among the handlers, partially based on the color of the clothes worn. This discrimination was generalized to another location. PMID- 9201581 TI - Evaluation of selected antibiotic residue screening tests for milk from individual goats. AB - Because somatic cell counts (SCC) of caprine milk are higher than SCC of bovine milk, the performance of antibiotic residue tests for screening bovine milk was investigated for caprine milk. Eighty-five does that were free of antibiotic usage for at least 30 d and that were free of clinical mastitis were sampled at three milkings during a 37-d period. At each sampling, foremilk was collected for bacteriological analysis, and composite bucket milk samples were collected for antibiotic testing and SCC. Day of lactation, parity, 305-d mature equivalent milk yield, and SCC averaged 221 d (57 to 577 d), 2.3 lactations (one to nine lactations), 1160 kg (623 to 1750 kg), and 2.2 x 10(6)/ml (0.3 to 30.7 x 10(6)/ml), respectively. The mean Dairy Herd Improvement Association test day milk yield for the month of sample collection was 3 kg (1.4 to 6.4 kg). Intramammary infections were present in 54% of the goats and in 36% of the udder halves. Assays included positive (5 and 10 ppb of penicillin-G and 50 ppb of ceftiofur) and negative controls that had been prepared in caprine milk and controls supplied by the manufacturers. One false-negative outcome and one false positive outcome were recorded. For one sampling day, a positive linear relationship existed between SCC and the results of one test, and a quadratic relationship existed between SCC and the results of another test. The antibiotic residue screening tests for milk from individual goats adequately identified milk that was free of antibiotic. These tests are therefore recommended for use with caprine milk. PMID- 9201582 TI - Effect of yeast culture (Saccharomyces cerevisiae) on adaptation of cows to diets postpartum. AB - For approximately 14 d prepartum and exactly 14 d postpartum, 20 multiparous Holstein cows were fed different basal diets that were supplemented, or not supplemented, with a yeast culture preparation. Cows supplemented with yeast culture lost less body condition prepartum, which was consistent with numerically higher body weight gain. No treatment differences were found in prepartum or postpartum dry matter intakes (DMI) or components of DMI. In addition, the extent of the depression in DMI prepartum and the rate of increase in DMI postpartum were not influenced by yeast culture supplementation. Milk and milk component yields were not influenced by yeast culture supplementation. Cows in both groups had higher calculated net energy for lactation for the diets postpartum than would have been expected based on values of the National Research Council for feedstuffs. The increased net energy for lactation seemed to be related to higher than expected metabolic efficiency during early lactation. Results of both the prepartum and postpartum periods were consistent with the hypothesis that supplementation of yeast culture in the diet increased net digestion in the forestomach, particularly of fiber, leading to increased energy output. However, there was no evidence to suggest that supplementation of yeast culture prepartum alleviated the reduction in DMI prepartum or improved the rate of increase in DMI postpartum. PMID- 9201583 TI - Effects of cellobiose and monensin on in vitro fermentation of organic acids by mixed ruminal bacteria. AB - The objective of this study was to determine the effects of cellobiose and monensin on the in vitro fermentation of organic acids (L-aspartate, fumarate, and DL-malate) by mixed ruminal bacteria. Ruminal fluid was collected from a steer fed 36.7 kg of forage and 4.5 kg of concentrate supplement once per day. Ruminal fluid was centrifuged to sediment feed particles and protozoa, and the resulting supernatant, which contained bacteria, was added (33%, vol/vol) to anaerobic media (500 ml). Incubations (n = 2) were performed in batch culture at 39 degrees C and sampled at 0, 2, 4, 6, 8, 12, and 24 h. Organic acids were added to achieve a final concentration of 7.5 mM. Cellobiose was added to obtain a final concentration of 5 mM, and monensin dissolved in ethanol was included at concentrations of 0 or 5 ppm. Addition of cellobiose to organic acid fermentations increased the rate of organic acid utilization by the mixed bacterial population. Total concentrations of volatile fatty acids were increased by the addition of cellobiose to all fermentations. A lag period (< or = 8 h) occurred in fermentations that were treated with monensin before organic acids were utilized. Total concentrations of volatile fatty acids were increased, and the acetate to propionate ratio was decreased, by monensin treatment. When cellobiose and monensin were added together, propionate production and organic acid utilization were increased. Both cellobiose and monensin affected the in vitro fermentation of organic acids by mixed ruminal bacteria by providing a carbon and energy source and by influencing electron disposal. PMID- 9201585 TI - Relationships between in situ protein degradability and grass developmental morphology. AB - The objective of this research was to determine the relationships between the morphological development and in situ ruminally degradable protein (RDP), ruminally undegradable protein (RUP), and microbial protein of two cool season grasses (intermediate wheatgrass and smooth bromegrass) and two warm season grasses (switchgrass and big bluestem). The initial growth of grass tillers grown near Mead, Nebraska was clipped at ground level six times during the 1992 growing season and morphologically classified. Mean stage was calculated. Forage was ground to pass a 2-mm screen and was incubated in ruminally fistulated steers for 16 h. The RUP was adjusted for microbial protein and acid detergent insoluble N. The mean stage of cool season grasses was higher than that of warm season grasses throughout the growing season. The RDP decreased as plant maturity increased for all species. The RUP expressed as a percentage of crude protein for the cool season grasses was lower than that for warm season grasses. The RUP for intermediate wheatgrass, smooth bromegrass, and switchgrass remained constant across maturities, but RUP for big bluestem decreased as maturity increased. Microbial augmentation of RUP decreased as crude protein decreased in all species. The RUP corrected for acid detergent insoluble N and microbial protein was relatively constant across plant maturities. The quantification of RUP across a range of plant maturities provided information for incorporating RUP content of forage grasses into the diets of animals. PMID- 9201584 TI - Effect of monensin on milk production and efficiency of dairy cows fed two diets differing in forage to concentrate ratios. AB - Four primiparous Holstein cows were gradually introduced, according to a Latin square design, to four diets obtained from the factorial combination of two forage to concentrate ratios (70:30 and 50:50) and two concentrations of monensin sodium (0 and 300 mg/d per cow). Addition of monensin tended to depress feed intake and milk fat content without affecting milk production and without interactions with forage to concentrate ratios. Ruminal propionate percentage was increased more by the addition of monensin to the low forage diet than by the addition of monensin to the high forage diet. Serum urea and concentrations of nonesterified fatty acids tended to decrease when monensin was added to the high forage diet but did not change when monensin was added to the low forage diet. The results suggested that monensin had moderate positive effects on efficiency of milk production and might have an antiketogenic effect with high forage diets. PMID- 9201586 TI - Effects of amount and ruminal degradability of protein on nutrient digestibility and production by cows fed tallow. AB - Five cows with ruminal cannulas were used in a 5 x 5 Latin square design to determine the effects of fat and amount and ruminal degradability of dietary crude protein (CP) on nutrient digestibility and production of milk and milk components. Treatments were 1) control; 2) 15% CP, soybean meal; 3) 15% CP, by product proteins; 4) 18% CP, soybean meal; and 5) 18% CP, soybean meal and by product proteins. Diets 2 through 5 contained 3.5% tallow. Diets consisted of 28% alfalfa haylage, 22% corn silage, and 50% concentrate on a dry matter (DM) basis. Fat did not affect dry matter intake or percentages and yields of fat and CP in milk but increased milk yield 2.5 kg/d. Fat did not affect N fractions in milk but decreased the percentages of short- and medium-chain fatty acids (C6:0 to C16:0) and increased the percentages of long-chain fatty acids (C18:0 and C18:1) in milk fat. Fat did not affect ruminal fermentation characteristics or the percentages of dietary DM, organic matter, CP, acid detergent fiber, neutral detergent fiber, starch, ether extract, and energy that were digested. An increase in dietary CP from 15 to 18% increased dry matter intake 1.7 kg/d; increased intake of gross energy 8 Mcal/d; increased the percentages and quantities of DM, organic matter, CP, and energy digested; increased the quantities of acid detergent fiber and neutral detergent fiber digested; decreased ruminal pH; increased concentrations of total volatile fatty acids; and increased NH3 N in ruminal fluid. However, the difference in dietary CP did not affect milk yield or composition. Replacement of soybean meal in the diet with a mixture of by-product proteins decreased NH3 N in ruminal fluid, tended to decrease concentrations of total volatile fatty acids and increase pH of ruminal fluid, but did not affect milk yield or percentages and yields of milk components. PMID- 9201587 TI - Effects on nutrient digestion of wheat processing and methods of tallow addition to the diets of lactating dairy cows. AB - Five multiparous Holstein cows in midlactation that were fitted with ruminal and duodenal cannulas were used in a 3 x 5 incomplete Latin square. The objective of this study was to examine the effects on nutrient digestion of wheat processing and method of tallow addition to the diets of lactating dairy cows. Diets consisted of 45% forage and 55% concentrate, and each diet contained 20% wheat and 2% tallow (as-fed basis). Treatments were dry-rolled wheat with tallow added to the concentrate, steam-rolled wheat with tallow added to the concentrate, and steam-rolled wheat with tallow added first to the wheat. The dry matter intake; digestion of starch, fiber, and fatty acids; ammonia N concentration; and molar proportions of volatile fatty acids in ruminal fluid were not affected by treatments. The apparent digestibility in the total tract of organic matter and nitrogenous compounds was significantly higher for the steam-rolled treatment with tallow added first to the wheat. Mean ruminal fluid pH was similar across treatments; however, cows fed the diet containing steam-rolled wheat with tallow added first to the wheat had the smallest pH change from 0 to 2 h postfeeding. Milk yield did not differ, regardless of cow diet. Method of tallow addition had marked effects on the apparent digestibility of organic matter and N in the total tract of lactating dairy cows. PMID- 9201588 TI - Effects of ruminally inert fat and evaporative cooling on dairy cows in hot environmental temperatures. AB - Under hot summer conditions of Tucson, Arizona, 24 Holstein cows (mean = 80 d of lactation) were assigned for 56 d to four treatments in a randomized block design with a 2 x 2 factorial arrangement of treatments. Factors were 1) medium [4.6% of dry matter (DM)] versus high (7.4% of DM) amounts of dietary fat and 2) corral shade only versus shade equipped with evaporative cooling. The high fat diet contained 3% prilled fatty acids. The efficiency of the conversion of feed to milk tended to be better for cows fed prilled fat than for cows fed medium dietary fat, but other lactation measurements were unaffected. Cows with access to evaporative cooling had greater milk yields than did cows with access to shade only. Prilled fatty acids did not depress the percentage of milk protein, but reduced short- and medium-chain fatty acids (C6:0 to C14:0) in milk fat and increased palmitic acid. Digestibilities of DM, organic matter, crude protein, acid detergent fiber, neutral detergent fiber, and starch were unaffected by amount of fat or by cooling method, but prilled fatty acids tended to decrease apparent digestibility of fatty acids. No differences were observed among treatments in respiration rates or rectal temperatures. When rectal temperatures were determined, cows were crowded, which probably negated detection of an effect of evaporative cooling. Evaporative cooling increased milk yield of cows in hot weather, but the addition of 3% fatty acids did not increase yield, and no interactions were observed. PMID- 9201589 TI - Influence of tallow and Aspergillus oryzae fermentation extract in dairy cattle rations. AB - Objectives were to determine the effects of adding 3 g/d of Aspergillus oryzae fermentation extract to diets with or without 5.6% added tallow. Twenty-eight Holstein cows (mean = 98 d of lactation) were assigned to a randomized block experiment in a 2 x 2 factorial arrangement of treatments. Treatments were the basal diet 1) without tallow or extract, 2) with extract but no tallow, 3) with tallow but no extract, and 4) with tallow and extract. Milk production, dry matter intake, 3.5% fat corrected milk, digestibility of neutral detergent fiber in the total tract were depressed for cows fed tallow. Addition of fermentation extract did not stimulate fiber digestion or milk production of cows fed diets with or without fat. Addition of extract did not overcome depression of fiber digestibility by cows fed tallow. PMID- 9201590 TI - Lactational response of cows to different concentrations of calcium salts of canola oil fatty acids with or without bicarbonates. AB - Holstein cows (n = 24) averaging 39 d of lactation were used in a randomized complete block design during an 8-wk trial. From wk 1 to 4, diets contained 62% alfalfa silage and 38% concentrates (dry matter basis), and, from wk 5 to 8, diets contained 47% forage and 53% concentrates. The concentrates were increased for the second phase so that the effect of bicarbonates could be expressed more fully. Diets 1, 2, and 3 contained 2% of a blend of Na and K bicarbonates and 0, 2, or 4% of Ca salts of canola oil fatty acids (percentage of dry matter), respectively. Diet 4 contained the same percentage of Ca salts as did diet 3 but without bicarbonates. Dry matter intake decreased linearly (wk 4), and milk yield was altered quadratically (wk 4), as the percentage of Ca salts in the diet increased. Milk fat percentage (wk 8) and yield (wk 4 and 8), as well as milk protein percentage (wk 4 and 8) and yield (wk 4), decreased linearly as the percentage of Ca salts in the diet increased. Short- and medium-chain saturated fatty acids decreased linearly, and C18:0, trans-delta-11-C18:1, cis-delta-9 C18:1, cis-delta-11-C18:1, and C18:2 increased linearly, as Ca salts in the diet increased. Addition of Na and K bicarbonates to the diets that contained Ca salts increased milk and milk protein yields and increased the proportions of C18:2 in milk fat at wk 8. Dietary bicarbonates had no effect on the responses of other milk fatty acids to supplementation of 4% Ca salts of canola oil fatty acids. PMID- 9201591 TI - Response of lactating cows to methionine or methionine plus lysine added to high protein diets based on alfalfa and heated soybeans. AB - Lactation diets based on wilted alfalfa silage and heated whole soybeans are common in the midwestern US. We examined the milk production response of multiparous Holstein cows to the addition of ruminally protected methionine at two percentages to a basal total mixed ration. An additional total mixed ration included both methionine and lysine supplementation. Sixteen Holstein cows in early lactation were used in a replicated 4 x 4 Latin square design with 21-d periods. Milk production, milk composition, and dry matter intake were determined for the last 5 d of each period. Milk production (41.5 kg/d), dry matter intake (25.9 kg/d), and milk fat concentration (3.26%) were unaffected by the supplementation of amino acids. The addition of methionine increased milk protein concentration and yield linearly. Each gram of methionine increased milk protein yield by 4 g, and milk protein concentration increased from 2.89 to 2.99% with the addition of 10.5 g/d of methionine. The proportion of casein N in total milk N was unaffected by treatment. The addition of lysine did not elicit a response. Total mixed rations based on alfalfa haylage, heated soybeans, and animal proteins were clearly limited by their methionine content but were adequate in their lysine content. PMID- 9201592 TI - Supplementation of nicotinic acid for lactating Holstein cows under heat stress conditions. AB - Twenty-six lactating Holstein cows (90 d of lactation) were blocked according to milk production, parity, and days of lactation for assignment to one of two dietary treatments. Diets included a control diet with no supplemental niacin and a diet supplemented with increasing concentrations of niacin (12, 24, or 36 g/d per cow over three consecutive 17-d periods. Cows were housed in a covered free stall barn and were fed and milked twice daily. Mean maximum air temperatures and temperature-humidity indexes were 28.5, 31.4, and 25.2 degrees C and 79.6, 85.1, and 75, respectively, for the three periods. Rectal temperature was measured with a rectal probe, tail and rump temperatures by infrared thermometry, and respiratory rate by visual observation. Measurements were made daily at 0800, 1600, and 2200 h. Rectal temperature was not affected by treatment. Comparison of skin temperatures for control cows and cows fed niacin showed higher temperatures at the tail (34.0 vs. 33.7 degrees C at 0800 h; 35.1 vs. 34.8 degrees C at 1600 h, respectively) and rump (34.1 vs. 33.7 degrees C at 0800 h; 35.3 vs. 35.0 degrees C at 1600 h, respectively) for control cows during period 1. No differences in thermal responses were observed during period 3. Niacin did not significantly increase milk production but decreased skin temperatures during periods of mild or severe heat stress. PMID- 9201593 TI - Dietary mixtures of sodium bicarbonate, sodium chloride, and potassium chloride: effects on lactational performance, acid-base status, and mineral metabolism of Holstein cows. AB - The objective of this study was to determine lactational, blood mineral, and blood acid-base responses to dietary mixtures of NaHCO3, NaCl, and KCl and dietary cation-anion difference by lactating diary cows. Three 100:0:0 (primary) blends, three 50:50:0 (binary) blends, and one 33:33:33 (tertiary) blend of NaHCO3, NaCl, and KCl, respectively, were formulated to replace 1% of the dry matter in a diet based on corn silage. Seven treatments were defined according to a simplex-centroid mixtures design using a partially balanced incomplete block arrangement. An eighth treatment served as a control and contained 1% SiO2 instead of the mineral blends. Dietary cation-anion difference ranged from +25 to +40 meq of (Na + K - Cl)/100 g of dietary dry matter. Diets were fed for three consecutive 28-d periods during summer to 36 midlactation cows. Cows that were fed the tertiary mixture had lower milk protein percentage, whole blood bicarbonate, and plasma K than did cows fed the other blends. With the exception of milk protein percentage and body weight gain, none of the mixtures had a significant impact on lactational performance. The lack of differences could have been due to the narrow range in the dietary cation-anion difference studied. PMID- 9201594 TI - Genetic evaluation of dairy cattle using test day yields and random regression model. AB - A model for analyzing test day records that contains both fixed and random regression coefficients was applied to the genetic evaluation of first lactation data for Canadian Holstein dairy cows. Data were 5.1 million test day records with milk, fat, and protein yields from calvings between 1988 and 1995 from herds in four regions of Canada. Each evaluated animal received five predictions for each trait representing the random regression coefficients. From these solutions, a range of estimated breeding values for various parts of the lactation could be calculated. Three genetic measures of persistency were compared. Bulls could be selected for both yields and persistency of their daughters in whatever combination was desirable. Test day analyses could result in monthly genetic evaluations for yield traits. PMID- 9201595 TI - Estimation of genetic variation in the interval from calving to postpartum ovulation of dairy cows. AB - Data on the interval from calving to the commencement of luteal activity of postpartum dairy cows were obtained for 1737 lactations of 1137 British Friesian cows in 11 commercial herds and 1 experimental herd between 1975 and 1982. The interval from calving to commencement of luteal activity was measured using progesterone concentrations of milk samples that were collected three or more times per week from shortly after calving to approximately 100 DIM of the following gestation. Genetic models were fitted using REML and accounting for known genetic relationships. Estimates of heritability and repeatability were 0.28 and 0.28, respectively, for the untransformed data; 0.21 and 0.26, respectively, for log-transformed data; and 0.13 and 0.26, respectively, after reciprocals were considered. In all cases, the heritability was significantly different from 0, and, of three scales, the log transformation had the greater likelihood. The likelihood of the transformation was closely related to the magnitude of the coefficient of skewness, and the power transformation with maximum likelihood was between 0.35 and 0.30, for which heritability was 0.19. The geometric mean interval was 25.6 d; coefficient of variation was 37%; and herds, years, parity, and season all had significant effects upon the interval to commencement of luteal activity. The postpartum interval grew longer by 2.2% with each parity [confidence interval 95% (1.1%, 3.0%)] and showed seasonal variation. Cows calving during spring took 1.21 times longer to commencement of luteal activity than did cows calving during autumn [95% confidence interval (1.13, 1.29)]. Genetic regression on PTA of the sire for milk, fat, and protein yields and for fat and protein percentages and on a national economic index were carried out using a subset of animals during 721 lactations. The regression was positive for fat percentage, but not significantly different from 0 for others. The magnitude of the heritability estimate in this study indicates that the postpartum interval to commencement of luteal activity may be useful for selecting cattle for improved fertility because shorter intervals have been postulated to be correlated with higher reproductive efficiency. PMID- 9201596 TI - Genetic markers in early diabetic retinopathy of adolescents with type I diabetes. AB - The aim of this study was to evaluate the role of HLA (human leucocyte antigen) class I (A, B, C) and class II (DR) alleles and familial insulin-dependent diabetes mellitus as possible risk markers for early retinopathy in a population of 103 Finnish adolescents with type I diabetes mellitus for 3.6-16.2 years. Fifty-one of the patients (49.5%) had signs of retinopathy in fundus photographs. HLA DR1 was found in 31% of the subjects with retinopathy, but in only 5% of those without retinopathy (p = 0.02). The corresponding figures for HLA DR1/4 were 17% and 2.6%, respectively (p = 0.22). The frequency of HLA DR3, DR4, or DR3/4 heterozygosity did not differ between the two groups of patients. Signs of early retinopathy showed thus an association with the presence of the HLA DR1 allele, and a mild protective effect of the HLA A9 and B40 alleles was indicated. Other HLA A, B, C, or DR alleles did not have any effect on the risk for early development of retinopathy, neither had a positive family history of type I diabetes. PMID- 9201598 TI - Early retinal and renal abnormalities in diabetes. AB - The integrity of the blood-retinal and blood-glomerular vascular barriers were investigated simultaneously in diabetic individuals to determine whether or not the early forms of diabetic retinopathy and nephropathy are temporally related. The blood-retinal barrier was assessed by the technique of vitreous fluorophotometry. Twenty-four hour urinary excretion of albumin was determined by radioimmunoassay before fluorescein measurement. Posterior vitreous fluorescein leakage was greater in the study cohort than in the control population after diabetes had been present 11-20 years (p < 0.05) and 21 years or more (p < 0.01). Albumin excretion was also increased in the diabetic subjects (p < 0.001) and correlated to duration of diabetes (r = 0.51, p < 0.005). Hypertension raised midvitreous fluorescein levels (p < 0.05), but it had no effect on posterior vitreous values. Hypertension was an independent predictive factor for urinary albumin excretion (p < 0.05). Partial correlation analysis showed that vitreous fluorescence and urinary protein were not significantly correlated when controlled for duration of diabetes and for age. Early proteinuria did not predict retinal vascular leakage, nor did increased fluorescein leakage predict renal decompensation in the diabetic subjects. The data suggest that during the early stages of retinal and renal abnormalities associated with insulin-dependent diabetes, the eye and kidney follow different temporal courses to abnormal function. PMID- 9201597 TI - Limited joint mobility in non-insulin-dependent diabetic (NIDDM) patients: correlation to control of diabetes, atherosclerotic vascular disease, and other diabetic complications. AB - This study examined the association between limited joint mobility (LJM) and diabetic control, atherosclerotic vascular disease and other diabetic complications in non-insulin-dependent diabetic (NIDDM) patients. LJM was studied in 139 [age (mean +/- SD) 61.3 +/- 12.3 years] NIDDM patients. Limitation of several joints was examined with a goniometer and LJM was classified by the Rosenbloom method. The NIDDM patients were examined for the following diseases: history of myocardial infarction, coronary heart, cerebrovascular and peripheral vascular diseases. The diabetic complications, background and proliferative retinopathy, nephropathy, and neuropathy, were also assessed. The metabolic control of the diabetes was evaluated by the average glycosylated hemoglobin Alc (GHbA kappa) concentration and lipid values were also measured. Mean levels of GHbAlc were 8.9 vs. 8.2% (p < 0.05) in NIDDM patients with and without LJM. NIDDM patients with LJM had a 3.1- (95% confidence interval, 1.2-7.7) and a 4.0-fold risk (95% confidence interval, 1.2-13.0) for coronary heart and cerebrovascular disease respectively, when the confounding effects of age, duration of diabetes and control of diabetes were controlled using stepwise logistic regression analysis. Patients with LJM had a 9.3- (95% confidence interval, 1.1-79.0) and a 3.3-fold risk (95% confidence interval, 1.0-10.5) of proliferative retinopathy and nephropathy respectively, when the confounding effects of age and duration of diabetes were controlled, but the correlation disappeared when control of diabetes was included in the model. In conclusion, the presence of LJM is associated with the control of diabetes and with the presence of coronary heart and cerebrovascular diseases in NIDDM patients. PMID- 9201599 TI - Measuring renal function in patients with diabetes mellitus. AB - We estimated the glomerular filtration rate in 33 patients from our diabetic clinic using three methods: the creatinine clearance measured from a timed urine sample and a serum creatinine; the creatinine clearance calculated from the serum creatinine according to the formula of Cockcroft and Gault; and, the plasma clearance of ethylenediaminetetra-acetic acid (EDTA) labeled with 51Cr ([51Cr]EDTA). We repeated the measurements in seven subjects. The measured creatinine clearance was not reproducible. The other two methods were correlated, but not according to the formula y = x. The calculated creatinine clearance significantly underestimated the [51Cr]EDTA clearance particularly at higher [51Cr]EDTA clearance rates. [51Cr]EDTA clearance has been shown by others to parallel, but underestimate, inulin clearance, the optimal method of estimating glomerular filtration rate but difficult to perform in routine practice. Accurately measuring renal function in routine clinical practice is difficult, and this must be borne in mind when making clinical decisions based on current measurements. PMID- 9201600 TI - Skeletal muscle lipoprotein-lipase activity in insulin-dependent diabetic patients with and without albuminuria. AB - In patients with insulin-dependent diabetes mellitus (IDDM), albuminuria reflects widespread vascular dysfunction. Albuminuria has been associated to defects of heparan sulfate proteoglycan (HSPG) within the extracellular matrix. Our hypothesis is that loss of HSPG in vascular walls reduces the HSPG-bound lipoprotein-lipase activity (LPLA), thereby causing elevated levels of plasma triglyceride (TG) seen in IDDM patients with albuminuria. The aim of the present study was to evaluate whether LPLA in muscle capillaries could be related to TG in IDDM patients with and without albuminuria. This is a cross-sectional study including ten healthy control subjects (group C), nine patients with IDDM and urinary albumin excretion rate (AER) of 30 mg/24 h or less (group D0) and 20 patients with IDDM and AER greater than 30 mg/24 h (group DA). Muscle LPLA, plasma TG, total cholesterol, high-density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL), and very-low-density lipoprotein cholesterol (VLDL) were measured. Between groups no difference in total cholesterol, TG, VLDL, and LDL was found. In patients with albuminuria, LPLA was reduced compared to controls, however, the difference between the groups was not statistically significant [median (range)] 35.9 mU/g (20.4-103) versus 44.6 mU/g (28.2-57.2) and 40.9 mU/g (21.7-53.5) in group DA, C, and D0, respectively, p = 0.76. AER was not correlated to LPLA. An overall negative correlation between TG and LPLA was found; r = -0.33, p = 0.04, supported by an overall significant positive correlation between LPLA and HDL; r = 0.32, p = 0.045. We conclude that, in insulin-dependent diabetes mellitus, skeletal muscle lipoprotein-lipase activity is associated with plasma triglyceride, while an association between lipoprotein-lipase activity and urinary albumin excretion is questionable. PMID- 9201601 TI - Dolichol-mediated enhanced protein N-glycosylation in experimental diabetes--a possible additional deleterious effect of hyperglycemia. AB - In liver cells from diabetic rats, an increased incorporation of labeled glucosamine into cellular and secretory proteins was found, when related to the incorporation of labeled leucine. This increased N-glycosylation was present in the face of decreased synthesis of hepatic cellular and secretory proteins evident from reduced leucine incorporation and diminished glycosyltransferase activity. To elucidate the mechanisms involved we incubated isolated hepatocytes with two N-glycosylation inhibitors: tunicamycin and 2-deoxyglucose. Tunicamycin exerted a marked inhibitory effect on the incorporation rate of labeled glucosamine into proteins in liver cells from diabetic rats, while 2-deoxyglucose had a negligible effect on this process in these cells. These diverse effects might be explained by the fact that tunicamycin acts through strong association with the enzyme catalyzing the first step in glycoprotein synthesis, namely, the transfer of UDP-GlcNAc to dolichol-P (indicating noncompetitive inhibition). This enzyme is reduced in liver cells from diabetic animals. On the other hand, 2 deoxyglucose exerts its effect by being attached to dolichol-P, preventing further elongation of oligosaccharide chain on the protein backbone. This latter effect might be eliminated by excess dolichol-P (indicating competitive inhibition). The dolichol content in liver extract from diabetic rats was about 2.5-fold higher compared with nondiabetic rats (51.6 micrograms/g versus 20.6 micrograms/g wet liver weight). These two lines of evidence confirm the notion that the enhanced enzymatic glycosylation in liver from diabetic animals is maintained by an increased hepatic dolichol concentration, which is most probably related to the hyperglycemia. Thus, the dolichol-N-glycosylation pathway may represent another detrimental aspect of hyperglycemia and may operate by dolichol mass action rather than through glycosylating enzyme activity. PMID- 9201602 TI - Do tissue plasminogen activator-plasminogen activator inhibitor-1 complexes relate to the complications of insulin-dependent diabetes mellitus? Pittsburgh Epidemiology of Diabetes Complications Study. AB - The purpose of this study was to examine the potential relationship of tissue plasminogen activator-plasminogen activator inhibitor-1 (tPA-PAI-1) complexes and diabetic complications in individuals with insulin-dependent diabetes mellitus (IDDM). To address this issue, data from the third follow-up visit of participants in the Epidemiology of Diabetes Complications (EDC) study were examined. There were 454 participants, aged 32 +/- 8 years, with duration of IDDM of 23 +/- 8 years. Higher levels of tPA-PAI-1 complexes were seen for both men and women with IDDM complications. Specifically, statistically significant differences were seen in men with neuropathy (1.81 +/- 0.9 versus 1.42 +/- 0.8 ng/mL, p < 0.01), microalbuminuria (1.77 +/- 1.1 versus 1.35 +/- 0.6 ng/mL, p < 0.01), retinopathy (1.67 +/- 0.9 versus 1.43 +/- 0.8 ng/mL, p < 0.05), and lower extremity arterial disease (1.93 +/- 0.7 versus 1.50 +/- 0.9 ng/mL, p < 0.05) versus men without the particular complication. In women, higher complex levels were shown for those with retinopathy (1.51 +/- 0.8 versus 1.29 +/- 1.1 ng/mL, p < 0.01). Potential mechanisms for the relationship of higher complex levels and diabetic complications include an altered fibrinolytic response and/or insulin resistance. Because the results are cross sectional, it cannot be established whether the higher concentration of complexes is a result of the presence of complications or are antecedent. Prospective follow-up will be required to determine if tPA-PAI-1 complexes are predictive of the development of IDDM complications. PMID- 9201603 TI - Hyperglycemia counterbalances the antihypertensive effect of glutathione in diabetic patients: evidence linking hypertension and glycemia through the oxidative stress in diabetes mellitus. AB - Diabetes mellitus is associated with hypertension. An antihypertensive effect of the antioxidant glutathione has been recently demonstrated. It has been suggested that hyperglycemia may contribute to the pathophysiology of hypertension in diabetes by generating an oxidative stress. In this study, three different tests were performed in ten hypertensive and ten nonhypertensive diabetic subjects: (1) an oral glucose tolerance test, (2) glutathione i.v. administration (1 g/m2 bolus + 1 g/m2 in 2 h), and (3) oral glucose tolerance test + glutathione administration. At -15', 0', 30', 60', 90', 120', and 180' systolic and diastolic blood pressure, plasma glucose, and insulin were measured. Variations in plasma glucose and insulin levels were not different during each test in the two groups of patients and in test (1) compared to (3). Glutathione administration reduced systolic and diastolic blood pressure in both hypertensive and nonhypertensive diabetic subjects from 30' to 120'. This phenomenon was abolished as glycemia increased after oral glucose loading. In hypertensive, but not in nonhypertensive diabetic subjects, a significant increase of systolic and diastolic blood pressure was observed at 90' and 120' of the oral glucose tolerance test (p < 0.01). These data show that hyperglycemia can counteract the hypotensive effects of the antioxidant glutathione, suggesting that glucose may impair arterial relaxation by producing free radicals. Also, it appears that hypertension in diabetic patients is aggravated by high glucose plasma levels. PMID- 9201604 TI - A pentamidine-treated acquired immunodeficiency syndrome patient associated with sudden onset diabetes mellitus and high tumor necrosis factor alpha level. PMID- 9201605 TI - The diabetic cataract: an unusual presentation in a young subject: case report. AB - This case report concerns a 14-year-old female patient, whose insulin-dependent diabetes mellitus was displayed by one infrequent complication, the cataract. This is an unusual manifestation in a 14-year-old patient; indeed, there are many findings in experimental animals demonstrating the development of this complication by maintaining blood glucose levels above 12 mM. After surgical therapy, complete vision was recovered, but we think that an earlier diagnosis and therapy of metabolic derangement of diabetes may have avoided this complication. PMID- 9201606 TI - Liquid chromatographic analysis of cocaine and benzoylecgonine in plasma of traditional coca chewers from Bolivia during exercise. AB - The purpose of the present study was to determine the amount of cocaine and benzoylecgonine in the plasma of Aymara Indians from the Bolivian Andes after traditional chewing of coca leaves during exercise performance. The determination was carried out by high performance liquid chromatography after solid-liquid extraction. The results showed that such use of coca leaves is well correlated with pharmacologically active concentration of cocaine in plasma. PMID- 9201607 TI - A study of the chemical composition of Erythroxylum coca var. coca leaves collected in two ecological regions of Bolivia. AB - Coca-Erythroxylum coca Lamarck var. coca-remains one of the most common plants of the folk medicine of Bolivia used as a general stimulant. Aymara and Quechua natives prefer to chew the sweeter coca leaves from the Yungas (tropical mountain forests of the eastern slopes of the Andes) rather than those from the Chapare lowlands. The contents in cocaine and minor constituents of leaf samples cultivated in these regions does not rationalize this choice. PMID- 9201608 TI - Hepatoprotective effect of the fractions of Ban-zhi-lian on experimental liver injuries in rats. AB - The hepatoprotective effect of various fractions (n-hexane, CHCl3, EtOAc, n-BuOH, and H2O) of Ban-zhi-lian derived from Scutellaria rivularis Benth was studied against carbon tetrachloride (CCl4), D-galactosamine (D-GalN) and acetaminophen (APAP)-induced acute hepatotoxicity in rats. Liver damage was assessed by quantifying serum activities of glutamate oxaloacetate transaminase (sGOT) and glutamate pyruvate transaminase (sGPT), as well as by histopathological examination. The results indicated that the CHCl3 fraction and EtOAc fractions exhibited the greatest hepatoprotective effects on CCl4-induced liver injuries, the CHCl3 fraction and n-hexane fraction are most potent against D-GalN-induced intoxication, and the CHCl3 fraction represented the most liver-protective effect on APAP-induced hepatotoxicity. The pathological changes of hepatic lesions caused by these three hepatotoxicants were improved by treatment with the fractions mentioned above, which were compared to Glycyrrhizin (GLZ) and Silymarin as standard reference medicines. PMID- 9201609 TI - Comparison between the essential oils of Santolina insularis (Genn. ex Fiori) Arrigoni and Santolina corsica Jord. et Fourr. from the island of Sardinia (Italy). AB - A comparative study of the essential oils of Santolina insularis and Santolina corsica has been carried out. The two specimens are found in Sardinia and were indiscriminately used in traditional medicine on the island. The essential oil was extracted by steam distillation of fresh aerial parts and analysed by GC, GC MS and GC-FTIR. The analysis of the essential oils shows substantial qualitative and quantitative differences. Some of the components identified were present in both investigated species, others were characteristic of one species only. PMID- 9201610 TI - The effect of Syzygium cumini (L.) skeels on post-prandial blood glucose levels in non-diabetic rats and rats with streptozotocin-induced diabetes mellitus. AB - This study was undertaken to investigate whether a tea prepared from Syzygium cumini, reported to be used by diabetics in Porto Alegre, Brazil, might have an antihyperglycemic effect in experimental models. Teas prepared from leaves and seeds of S. cumini, in concentrations ranging from 2-64 g/l, were administered, as water substitute for 14-95 days, to 16 groups with 8-9 normal albino rats and to four groups with 10-12 rats with streptozotocin-induced diabetes mellitus. Post-prandial blood glucose levels were determined by the glucose oxidase method on blood samples obtained by decapitation. None of the tea concentration had any detectable antihyperglycemic effect either in normal or in diabetic rats, suggesting that this plant, prepared in a manner similar to that employed by humans, is destitute of an antihyperglycemic effect. PMID- 9201611 TI - Effect of Maharishi AK-4 on H2O2-induced oxidative stress in isolated rat hearts. AB - Oxidative damage to crucial biomolecules due to excess generation of reactive oxygen species has been implicated as a major cause of organ damage and hence compounds capable of negating such damage have potential benefits. Using hydrogen peroxide (H2O2) as a model pro-oxidant to induce oxidative stress, we have examined the ability of natural food supplement Maharishi Amrit Kalash (MAK-4) to decrease oxidative damage in potassium-arrested isolated rat hearts. The protocol was that hearts isolated from male Sprague-Dawley rats were retrograde-perfused with Krebs-Henseleit (K-H) solution for 30 min for equilibration. After this period, the hearts were subjected to cardioplegia with high potassium (26-30 mM), followed by reperfusion with K-H solution in the presence or absence of 200 microM H2O2. As expected, H2O2 treatment following cardioplegia induced a high degree of oxidative stress as assessed by release of lactate dehydrogenase (LDH, a marker of plasma membrane damage) and total glutathione (GSH + GSSG). H2O2 also impaired the ability of heart to regain developed tension during the testing period. However, addition of MAK-4 in the perfusate containing H2O2 decreased oxidative stress in terms of release of LDH and glutathione. In parallel with these biochemical studies, in a few experiments the cardiac function was assessed by measuring developed contractile tension. These preliminary studies also showed that in the presence of MAK-4 the H2O2-treated hearts were able to regain better developed tension. Further in vitro studies to examine the possible mechanisms of MAK-4 action reveal that this formulation contains H2O2 binding activity which resulted in the decreased availability of H2O2 itself. Our studies hence reveal that the ayurvedic food supplement MAK-4 may have potential benefits in reducing oxidative stress. PMID- 9201612 TI - Chemical composition and antimicrobial activity of Croton urucurana Baillon (Euphorbiaceae). AB - In the methanolic extract of Croton urucurana Baillon (Euphorbiaceae) a number of known compounds, such as acetyl aleuritolic acid, stigmasterol, beta-sitosterol, campesterol, beta-sitosterol-O-glucoside, sonderianin, catechin and gallocatechin were isolated and identified by MS and NMR spectroscopy, HRGC and data from literature. The antibacterial activity of the aqueous-EtOH extract, some fractions of the methanolic extract and some of the isolated compounds, were tested against Staphylococcus aureus and Salmonella typhimurium. Acetyl aleuritolic acid exhibits the best minimum inhibitory concentration (MIC) against both Staphylococcus aureus and Salmonella typhimurium. PMID- 9201613 TI - Antimicrobial activity of flavonoids from leaves of Tagetes minuta. AB - The total extract and fractions with different solvents, obtained from leaves of Tagetes minuta, showed several degrees of antimicrobial activity against Gram positive and Gram negative microorganisms. The same fractions were inactive against Lactobacillus, Zymomonas and Saccharomices species. The major component of the extract: quercetagetin-7-arabinosyl-galactoside, showed significant antimicrobial activity on pathogen microorganisms tested. Correlation results were carried out using chloramphenicol as standard antibiotic. PMID- 9201614 TI - Management of giardiasis by a herbal drug 'Pippali Rasayana': a clinical study. AB - Pippali Rasayana (PR), an Indian ayurvedic drug prepared from Palash (Butea monosperma (Lamk) Kuntze; Leguminaceae) and Pippali (Piper longum L.; Piperaceae), was administered at a dose of 1 g p.o. three times daily for a period of 15 days to patients (25 treated, 25 placebo controls) suffering from giardiasis with clinical signs and symptoms, and stools positive for trophozoites/cysts of Giardia lamblia. After 15 days of drug treatment there was a complete disappearance of G. lamblia (trophozoites/cysts) from the stools of 23 out of 25 patients. General signs and symptoms of ill health and abdominal discomfort, presence of mucus, pus cells and RBCs were significantly reduced. There was a marked improvement in the clinical and haematological profile of the patients. Spontaneous recovery in 20% cases was recorded in placebo controls. PMID- 9201615 TI - Matching Ca efflux and influx to maintain steady-state levels in cultured cardiac cells. Flux control in the subsarcolemmal cleft. AB - The study examines the feed-back mechanism by which Ca efflux via Na/Ca exchange adjusts to match Ca influx via Ca channels-the condition which must obtain if cellular steady-state Ca level is to be maintained. It is proposed that variation in Ca channel influx produces changes in sarcoplasmic reticulum content which, in turn, through interaction within the subsarcolemmal cleft space, sets the level of Na/Ca exchange-mediated efflux. We measured Ca channel influx with whole-cell voltage-clamp, Ca efflux specifically via Na/Ca exchange by high resolution, non perfusion-limited 45Ca wash-out technique and sarcoplasmic reticulum (SR) Ca content by combination of the latter with measurement of sarcolemmal Ca-binding by use of instantaneous "gas-dissected" sarcolemmal membrane isolation. Ca channel influx was varied by Bay K 8644 or nifedipine. Ca influx via Ca channels was increased by 37.8% with Bay K and decreased by 68% with nifedipine. The SR contribution to Na/Ca exchange-mediated efflux was increased by 36.7% with Bay K and reduced by 66.8% by nifedipine-virtually identical changes. Application of the cleft space model to determine beat-to-beat efflux via Na/Ca exchange predicted values which closely matched beat-to-beat channel influx. PMID- 9201616 TI - Determinants of the S-adenosylhomocysteine (SAH) technique for the local assessment of cardiac free cytosolic adenosine. AB - The S-adenosylhomocysteine (SAH) technique allows the estimation of the free cytosolic adenosine concentration using the kinetic properties of the enzyme SAH hydrolase (adenosine+homocysteine reversible SAH+H2O). Besides the cytosolic adenosine concentration, the local SAH signal may also depend on the local homocysteine availability, the continuous production of SAH from S adenosylmethionine (SAM-->SAH+CH3) and the activity of the enzyme SAH-hydrolase. These variables were studied with high spatial resolution (sample dry mass 25 mg) in left ventricular myocardium from 26 anesthetized open-chest dogs in which heart rate averaged 86 +/- 14 beats/min and mean aortic pressure 96 +/- 17 mmHg. Homocysteine infusion (48 mg/kg i.v.) increased the normal plasma homocysteine concentration from 5.0 +/- 0.8 to 586 +/- 40 microM after 30 min when the average tissue concentration was 94% of the plasma concentration and similar in low and high flow areas (flow range 0.04 to 1.91 ml/min/g). Local SAH content was 1.18 +/ 0.48 nmol/g under control conditions and increased to 4.33 +/- 0.59 nmol/g within 60 min following competitive blockage of the SAH-hydrolase by adenosine dialdehyde (10 mumol/kg i.v.). This increase of the SAH content was slightly more in high than in low-flow areas (P < 0.01). Regional SAH-hydrolase activity (9.0 +/- 0.5 nmol/min/g) was comparable in high and low flow areas. All three variables exhibited an observed variability which was larger than the methodical variability suggesting significant spatial heterogeneity in the myocardium. A regrouping analysis indicated that between four and five samples taken from distant sites should be averaged to obtain a robust estimate of the above metabolic parameters. Reconciling the measurements with a mathematical model of cardiac adenosine metabolism and fitting of the measured SAH tissue levels gave an estimate of 72 pmol/min/g for the mean transmethylation rate. Estimates of the cytosolic adenosine concentration of cardiomyocytes and endothelial cells under control physiological conditions were 24 and 7 microM, respectively. Thus, the present measurements provide a basis for the quantitative assessment of the local cytosolic adenosine concentration in relation to blood flow. PMID- 9201617 TI - Primary culture of human atrial myocytes is associated with the appearance of structural and functional characteristics of immature myocardium. AB - We examined changes in the structural and physiological characteristics of human atrial myocytes during primary culture in the presence of serum. Action potentials and ionic currents were recorded in freshly dissociated (FM) and cultured (CM) whole-cell patch-clamped myocytes, alpha-smooth muscle actin, sarcomeric alpha-actinin and beta-myosin heavy chains (beta-MHC) were stained with monoclonal antibodies. From day 5 to day 21, myocytes lost their rod shape, spread and exhibited reorganized sarcomeres. These morphological changes were associated with a marked increase in membrane capacitance (+266%). Both beta-MHC and alpha-smooth muscle actin were expressed in CM but not in FM, indicating a dedifferentiation process. CM were characterized by a lower resting potential ( 30 +/- 2 v -60 +/- 4 mV, P < 0.05) and, when repolarized, by a shorter action potential duration (APD) than FM (APD-60: 126.9 v 159.6 ms, P < 0.05). The inward rectifier K+ current was absent in CM, thus explaining the low resting potential. The density of the transient component of the voltage-activated K+ current Ito1 was not modified during culture, while that of the sustained component Isus was increased fourfold. The amplitude of ICa was increased, but its density was unchanged, indicating that CM maintained a normal density of functional calcium channels. Neither the voltage dependence nor the inactivation of ICa was modified in CM. The time constants of inactivation of ICa were unchanged, although the amplitude of the rapidly inactivating component of ICa was increased in CM compared to FM. Moreover, ICa was increased by the beta-adrenergic agonist isoproterenol (1 microM) throughout the culture period. Our results demonstrate that in long-term serum-supplemented culture, adaptation of human atrial myocytes to their new environment is associated with differential alterations of the main ionic currents and phenotypic changes characteristic of immature myocardium. PMID- 9201618 TI - Vascular endothelial growth factor inhibits endothelial cell apoptosis induced by tumor necrosis factor-alpha: balance between growth and death signals. AB - A series of experiments was performed to determine whether vascular endothelial growth factor (VEGF), in addition to its endothelial cell specific mitogenic activity, can also protect endothelial cells from toxin-induced programmed cell death. Apoptosis was induced in endothelial cell culture with tumor necrosis factor-alpha (TNF-alpha). Simultaneous exposure of endothelial cells to VEGF resulted in a dose dependent inhibition of apoptosis when evaluated by: (1) direct counting of cells with morphologic features of apoptosis after acridine orange staining; (2) analysis of DNA fragmentation by (a) agarose gel electrophoresis and (b) fluorescence activated cell sorting (FACS); and (3) viability assays dependent upon mitochondrial function. Induction of fibronectin and beta 3 integrin expression in endothelial cells by VEGF suggests that altered adhesion molecule expression may explain this survival effect. PMID- 9201619 TI - Restoration of action potential duration and transient outward current by regression of left ventricular hypertrophy. AB - The presence of left ventricular hypertrophy (LVH) is associated with an increased incidence of arrhythmias. Our previous study on hypertrophied rat hearts has demonstrated that regression of LVH prevents ischemia-induced lethal arrhythmias. To elucidate the underlying mechanism of the reduced incidence of arrhythmias in regression of LVH, we examined electrophysiological properties of both hypertrophied and regressed left ventricular cells. Hearts from spontaneously hypertensive rats (SHR) were used as LVH, and those from Wistar Kyoto rats (WKY) served as control. SHR with regression of LVH (REG) was produced by captopril treatment. Action potentials and membrane currents of subendocardial left ventricular cells were compared by the whole-cell patch-clamp techniques. Although the membrane capacitance of SHR cells was significantly greater than that of WKY cells, that of REG cells was normalized to the control level. Prolonged action potential duration (APD) and reduced density of transient outward current (ito) in SHR cells was normalized by LVH regression (APD at 75% repolarization (ms) and ito density at +60 mV (pA/pF): WKY 36.1 +/- 4.2, 11.9 +/- 1.3, SHR 73.1 +/- 12.9, 5.2 +/- 0.7, REG 29.5 +/- 3.9, 10.4 +/- 2.0, P = 0.015, P = 0.001 v WKY). No significant differences were observed in the densities of steady-state outward current, inward rectifier current, and L-type Ca2+ current. The restoration of ito density by regression of LVH could normalize the prolonged APD in hypertensive LVH, which may be causally related to the reduced incidence of arrhythmias in LVH regression. PMID- 9201620 TI - Norepinephrine pretreatment attenuates Ca2+ overloading in rat trabeculae during subsequent metabolic inhibition: improved contractile recovery via an alpha 1 adrenergic, PKC-dependent signaling mechanism. AB - The present study was designed in order to investigate more precisely the role of calcium homeostasis maintenance in protein kinase C (PKC) mediated preconditioning. We used a 15 min pre-incubation period, with 1 mumol/l exogenous norepinephrine (NE) to pharmacologically precondition isolated, superfused rat trabeculae against contractile dysfunctioning following 120 min of metabolic inhibition (MI, in 2 mmol/l CN- containing Tyrode without glucose at 1 Hz stimulation frequency). Contractile recovery was studied during a subsequent 60 min recovery period (RP, in glucose containing Tyrode at 0.2 Hz). Tyrode was gassed with 95%, O2/ 5% CO2 and kept at a constant temperature of 24 degrees C. Force and intracellular free calcium ([Ca2+]i) were monitored throughout the experimental protocol; [Ca2+]i was measured using fura-2. Pretreatment with NE (group NE-I) significantly increased the fraction of trabeculae that resumed to contract during RP, from 36 +/- 13% (mean +/- S.E.M.) in controls to 82 +/- 10% (P < 0.05). In correspondence with this, NE-pretreatment increased the proportion of trabeculae in which the Ca2+ rise from the onset of rigor development during MI was attenuated. After 40 min of MI [Ca2+]i in the failing control, as well as failing group NE-I, trabeculae (1.08 +/- 0.20 and 1.51 +/- 0.26 mumol/l, respectively) was increased significantly compared to the mean value registered in the recovering preparations of these groups (0.34 +/- 0.04 mumol/l: P < 0.05). Specific inhibition of PKC with 2 mumol/l chelerythrine (group NE-IV) almost completely blocked the protection induced by NE-pretreatment, including its protective action against Ca2+ overload, i.e. the fraction of trabeculae that resumed to contract during RP returned to untreated control level (46 +/- 11%: P < 0.05 v group NE-I). Also in this case [Ca2+]i in the failing group NE-IV trabeculae after 40 min of MI was increased substantially, compared to the value measured in the recovering preparations (4.75 +/- 1.00 and 0.60 +/- 0.08 mumol/ l, respectively). The relative importance of both alpha-adrenergic and beta adrenergic receptor pathways in this preconditioning-like effect of NE pretreatment, was investigated using specific blockers. The results point to an alpha 1-adrenergic receptor mediated signaling mechanism, which enhances PKC dependent control of [Ca2+]i from the onset of rigor development during MI. PMID- 9201621 TI - Ischemic preconditioning is mediated by a peripheral opioid receptor mechanism in the intact rat heart. AB - Previously, our laboratory has shown that opioid receptors are involved in ischemic preconditioning (PC) in the intact rat heart; however, it is not known whether this cardioprotection is mediated by central or peripheral mechanisms. To test this hypothesis, both naloxone (NL), the non-selective opioid receptor antagonist and naloxone methiodide (QNL), its quaternary derivative which does not cross the blood-brain barrier, were used to determine if opioid receptor induced myocardial protection occurs via a central or peripheral locus of action in inactin-anesthetized, open-chested, Wistar rats. In group I, the control group was subjected to 30 min of occlusion and 2 h of reperfusion. In group II, ischemic PC was elicited by three 5-min occlusion periods interspersed with 5 min of reperfusion. In group III, QNL (10 mg/kg, i.v.) was administered 10 min before the 30 min of occlusion. Groups IV and V consisted of a dose-response effect of QNL on ischemic PC in which QNL (0.3 or 10 mg/kg, i.v., respectively) was given 10 min prior to ischemic PC. In addition, in groups VI and VII, one of two doses of naloxone (1 or 3 mg/kg, i.v.) was administered 10 min before ischemic PC. Infarct size (IS) as a percentage of the area at risk (AAR) was determined by tetrazolium staining. Ischemic PC reduced IS to 9 +/- 2% (P < 0.05) v control (53 +/- 4%). The low dose of QNL partially blocked the cardioprotective effect of ischemic PC; whereas the high dose completely abolished its cardioprotective effect. The high dose of QNL had no effect on IS alone. Similarly, the low dose of NL did not antagonize the cardioprotective effect of ischemic PC; however, the high dose completely abolished ischemic PC. These results indicate that the cardioprotective effect of ischemic preconditioning is mediated by a peripheral opioid receptor mechanism in the intact rat heart. PMID- 9201622 TI - Effect of ischemia on the fraction of ryanodine-sensitive cardiac sarcoplasmic reticulum. AB - The effect of 15 min of global, normothermic ischemia on cardiac sarcoplasmic reticulum (SR) was investigated using the Ca2+ uptake rate and 3H-ryanodine binding of ventricular homogenates and isolated SR vesicles. Ischemia did not affect ryanodine binding in the homogenate, while it increased it in the isolated SR vesicles. Although ischemia decreased the homogenate oxalate-supported Ca2+ uptake rate, measured in the presence of high ryanodine to close the ryanodine sensitive efflux pathway (+RY), its decrease of the Ca2+ uptake rate, measured in the absence of ryanodine (-RY), was more marked. This finding was also observed in the isolated SR. Although inhibition of the Ca-ATPase and its coupled Ca2+ uptake by thapsigargin proportionately decreased SR Ca2+ uptake -RY and +RY, ischemia decreased the Ca2+ uptake -RY proportionately more. This result suggested that there was a greater fraction of Ca2+ uptake activity in ryanodine sensitive vesicles after ischemia. However, ischemia also reduced the yield of SR activity in the isolated SR fraction and the results could potentially be due to differential selection of ryanodine-sensitive and ryanodine-insensitive SR in the isolation procedure. We directly tested the hypothesis that ischemia changes the fraction of Ca2+ uptake activity in the ryanodine-sensitive vesicles by estimating the Ca-oxalate capacity measured +RY and -RY. Ischemia decreased the capacity -RY much more than +RY in the homogenate, indicating that more of the SR volume and Ca2+ uptake activity was in the ryanodine-sensitive vesicles after ischemia. PMID- 9201623 TI - Cultured myofibroblasts generate angiotensin peptides de novo. AB - Scar tissue found at the site of myocardial infarction (MI) contains phenotypically transformed fibroblast-like cells termed myofibroblasts (myoFb). In injured cardiac tissue, autoradiography and immunolabeling have localized high density angiotensin (Ang) converting enzyme (ACE) and Ang II receptor binding to these cells, suggesting that they may regulate local concentrations of Ang II and transduce signals at this site. Ang II is known to modulate type I collagen gene expression of fibroblasts and myoFb, and to promote fibrous tissue contraction, each of which may contribute to tissue repair. It is unknown whether myoFb themselves generate Ang peptides de novo via expression of angiotensinogen (Ao), an aspartyl protease needed to convert Ao to Ang I, and ACE. We therefore isolated and cultured myoFb from 4-week-old scar tissue of the adult rat left ventricle with transmural MI. In cultured myoFb we found: (a) immunoreactive membrane-bound ACE, cytosolic cathepsin D (Cat-D), and AT, receptors by immunofluorescence and confocal microscopy, (b) mRNA expression for Ao, ACE, and Cat-D, but not renin, by reverse transcriptase-polymerase chain reaction, (c) production of Ang I and II in serum-free culture media; (d) absence of renin activity; (e) a time-dependent conversion of Ao to Ang I by myoFb cytosol, which was inhibited by pepstatin A, but not by renin inhibitor; and (f) significant increase in Ang II production (P < 0.05) by exogenous Ao and Ang I (10 nM), which was significantly blocked by lisinopril (0.1 microM: P < 0.05). Thus, cultured myoFb express requisite components and are able to generate Ang I and II de novo. In an autocrine and/or paracrine manner, Ang II may regulate myoFb collagen turnover and fibrous tissue contraction. PMID- 9201624 TI - Beta-adrenergic receptor signalling in stunned myocardium of conscious pigs. AB - The primary goal of this study was to compare the effects of isoproterenol which stimulates beta-adrenergic receptors and forskolin, and NKH 477, a water soluble derivative of forskolin, which stimulate adenylyl cyclase in stunned myocardium of conscious pigs, previously instrumented for measurements of left ventricular pressure and dP/dt, arterial pressure, and wall thickening. Ten min of coronary artery occlusion induced transmural reductions in blood flow (radioactive microspheres) in subepicardium (-98 +/- 2%) and subendocardium (-99 +/- 1%). Wall thickening (piezoelectric crystals) fell from 2.50 +/- 0.26 mm to -0.26 +/- 0.26 mm and remained depressed at 1.37 +/- 0.19 mm after 20-30 min coronary artery reperfusion, reflecting myocardial stunning. At that time, isoproterenol (0.2 microgram/kg) increased wall thickening in stunned myocardium (+1.40 +/- 0.16 mm, P < 0.05) more than in control (+0.71 +/- 0.22 mm), while forskolin elicited the opposite effects. NKH 477 (30 micrograms/kg), which does not penetrate the blood brain barrier, increased systolic wall thickening similarly before (+0.95 +/- 0.25 mm) and during (+1.01 +/- 0.24 mm) myocardial stunning. The reflex inotropic responses to inferior vena caval occlusion on wall thickening were diminished, P < 0.05, in the stunned myocardium (+0.53 +/- 0.05 mm) compared with control (+0.95 +/- 0.07 mm). beta-adrenergic receptor density, which was quantitated with 125I-cyanopindolol binding, was increased transmurally in stunned myocardium compared with non-ischemic myocardium (subepicardium: +23 +/- 5%, subendocardium: +34 +/- 13%, P < 0.05). Basal and forskolin-stimulated adenylyl cyclase activities were decreased slightly, but significantly, in the stunned subendocardium but not in the subepicardium, while isoproterenol stimulation of adenylyl cyclase activity showed no differences. In summary, paradoxical responses to beta-adrenergic receptor stimulation were observed in stunned myocardium, with pharmacological stimulation with isoproterenol evoking enhanced responses, and neural stimulation with inferior vena caval occlusion eliciting depressed responses. The diminished responses to forskolin in vivo, in stunned myocardium were out of proportion to the biochemical measurements, and may be attributed to neurally mediated cardiac effects of forskolin, since the responses to direct stimulation of adenylyl cyclase by NKH 477 were preserved. PMID- 9201625 TI - Phospholipid degradation in energy-deprived cardiac myocytes: does annexin V play a role? AB - This study explored whether annexin V, a protein with established phospholipase A2 inhibiting properties, plays a role in the degradation of membrane phospholipids of adult cardiac myocytes during metabolic inhibition (20 mM 2 deoxyglucose and 1 mM iodoacetic acid). Experiments were carried out on isolated cardiac myocytes prelabeled with [14C]-arachidonic acid, which were subjected to metabolic inhibition for up to 240 min. Under control conditions, annexin V was found to be localised predominantly at the sarcolemma. After 120 min of metabolic inhibition, the release of lactate dehydrogenase (LDH) was still comparable with control cells, while morphological changes were already visible. After 240 min of metabolic inhibition, LDH release was significantly elevated compared to control cells incubated for the same period of time (35% v 20% of total cellular activity). All myocytes had lost their typical elongated shape and sarcolemmal "blebs" had been formed. In metabolically inhibited cells, annexin V localisation seemed to be more pronounced at the level of the sarcolemma compared to controls, whereas membrane phospholipid hydrolysis occurred at a significantly elevated rate, as evidenced by a significantly enhanced accumulation of labeled arachidonic acid within the cells. The present findings are not in favor of the hypothesis that the increase in net degradation of phospholipids in energy deprived cardiac myocytes is caused by a loss of annexin V from the sarcolemma, which would increase the vulnerability of the sarcolemma to phospholipase A2 activity. PMID- 9201626 TI - Cyclic nucleotides regulate the activity of L-type calcium channels in smooth muscle cells from rat portal vein. AB - In order to advance our previous findings that the macroscopic slow Ca2+ currents of vascular smooth muscle (VSM) cells are regulated by cyclic nucleotides, the effects of cAMP and cGMP on the activity of single slow (L-type) Ca2+ channels were investigated using cell-attached patch clamp (22-25 degrees C). Freshly isolated VSM cells were obtained from adult male rat portal vein. For the single channel recordings, the pipette was filled with a solution containing 90 mM Ba2+ and 1 microM Bay-K-8644 solution, and the bath contained 140 mM KCl to "zero" the membrane potential. Depolarizing pulses to 0 mV, from a holding potential (HP) of -80 mV, elicited inward unitary currents. The activity of these channels was completely blocked by superfusion of 10 microM nifedipine. Extracellular perfusion of the single cells with membrane-permeable cGMP and cAMP analogs (8Br cGMP and 8Br-cAMP) at 1 mM caused a slight inhibition, but higher doses (3 mM), clearly showed an inhibitory effect on the single-channel activity. cAMP (100 microM) stimulated one out of five patches tested, and 100 microM cGMP showed no effect in three patches tested. Compared with control, both cyclic nucleotides at 3 mM decreased the ensemble-averaged currents by 26.7 +/- 4.1% and 37.3 +/- 2.1%, respectively. Unit amplitude and slope conductance were not changed. The normal conductance of the Ca2+ channel was 20.8 +/- 0.04 pS (n = 9), and the conductances in the presence of cAMP (n = 5) and cGMP (n = 6) were 19.3 +/- 0.04 and 20.5 +/- 0.05 pS, respectively. Single-channel kinetic analysis showed that cAMP did not affect the mean open-time, and cGMP slightly decreased the mean open time. However, both cAMP and cGMP increased the mean closed-time. In addition, cAMP decreased the open probability (NPo) by a factor of 1.7, from 0.26 +/- 0.04 to 0.15 +/- 0.03 (P < 0.05, Student's t-test) and cGMP decreased NPo by a factor of 2.5, from 0.24 +/- 0.08 to 0.10 +/- 0.02 (P < 0.05). H-7, a non-specific protein kinase inhibitor, prevented the inhibitory effects of both cAMP and cGMP on the activity of single Ca2+ channels in rat portal vein cells. The results demonstrate that both cAMP and cGMP inhibit L-type Ca2+ channel activities in VSM cells from rat portal vein. This inhibition may be mediated by the cAMP and cGMP dependent protein kinase phosphorylation of the L-type Ca2+ channels (or an associated regulatory protein). PMID- 9201627 TI - Lack of evidence for connexin 43 gene mutations in human autosomal recessive lateralization defects. AB - Heterotaxy is the failure of the developing embryo to establish normal left-right asymmetry, which is often associated with multiple malformations. Previous studies have identified different mutations in the cytoplasmic tail of the connexin 43 (cx 43) gene in six patients from a series of six sporadic cases with defects of laterality and severe heart malformations. These cases showed that of the genes involved in lateralization defects with autosomal recessive transmission, cx 43 was the most important. This result was challenged by two different teams, which, on sequencing only the carboxyl terminal end of the cx 43 gene in 30 patients, found no mutations. To assess the responsibility of the cx 43 gene in human autosomal recessive lateralization defects, we tested its involvement in a selected group of 25 patients (19 familial cases) with a wide variety of lateralization defects and cardiovascular malformations. The whole coding sequence and direct flanking sequences were screened for mutations, both by single strand conformation analysis and direct fluorescent sequencing. We could only detect a single base pair insertion in the 3' untranslated region of one patient. To test the possibility of mutations in other parts of the cx 43 gene, the gene was located onto the physical map of chromosome 6, and flanking polymorphic markers were genotyped. Haplotype analysis excluded the cx 43 gene locus in nearly all of the familial cases of lateralization defects. Thus, our results do not support the suggestion that this gene is implicated in human autosomal recessive lateralization defects. PMID- 9201628 TI - Mechanisms of suppression and initiation of pacemaker activity in guinea-pig sino atrial node superfused in high [K+]o. AB - The electrophysiological mechanisms by which changes in [K+]o suppress and initiate pacemaker activity were studied in guinea-pig isolated sino-atrial node (SAN) superfused in vitro. High [K+]o (10 mM or higher) gradually decreases maximum diastolic potential and action potential amplitude, until only subthreshold responses and eventually quiescence follow. When the threshold potential is missed, an oscillatory afterpotential (Vos) is often superimposed on early diastolic depolarization (DD). During the subsequent late DD, gradually increasing oscillatory prepotentials (ThVos) appear, whose depolarizing phase may initiate an action potential. If ThVos miss the threshold, they gradually decrease in size. In quiescent SAN, on decreasing high [K+]o, the resumption of spontaneous activity is caused by ThVos. In high [K+]o, Cs+ and Ba2+ may induce spontaneous activity in quiescent SAN and accelerate spontaneously active SAN. A low [Ni2+]o does not suppress SAN, whereas nifedipine blocks excitation (but not DD); and high [Ca2+]o induces spontaneous discharge in quiescent SAN. Tetrodotoxin and low [Na+]o often cause block of conduction. In conclusion, high [K+]o suppresses SAN discharge not by abolishing DD, but by preventing the attainment of the threshold. A slower rhythm may be maintained by ThVos arising during the late DD. After arrest, resumption of activity is due to gradually increasing ThVos. The effects of current blockers suggest that in high [K+]o the mechanism underlying DD may involve IK, but not I(f) or ICa. Initiation of discharge by high [Ca2+]o and induction of quiescence by nifedipine suggest a role of Ca2+ in excitation (but not in DD). The effects of tetrodotoxin and low [Na+]o suggest a role of Na+ in conduction within SAN superfused in high [K+]o. PMID- 9201629 TI - Epinephrine facilitates cardiac fibrillation by shortening action potential refractoriness. AB - Epinephrine released during ventricular tachycardia (VT) or early fibrillation (VF) appears to be instrumental in stabilizing fibrillation. However, mechanisms remain unclear. Effects of epinephrine on refractory period at normal sinus rates depend on basic cycle length, but effects at short cycle lengths, typical of VT/VF, are unknown. Therefore, the goal of this study was to determine whether epinephrine shortens action potential duration and refractoriness at these short cycle lengths. To simulate early VT/VF, myocardial cell aggregates (n = 35) were paced using field stimulation (5 ms rectangular waveform) at cycle lengths of 200, 180, 160 and 140 ms, which occur during in situ fibrillation: normal sinus rhythm was simulated by pacing at 600 and 400 ms intervals. Action potentials and excitation threshold were recorded with intracellular microelectrodes under control conditions, with 0.9 microM/l epinephrine, and with 0.9 microM/l epinephrine and 0.5 microM/l propranolol. At short cycle lengths, epinephrine significantly shortened action potential duration and refractoriness compared to control. At a cycle length of 160 ms, action potential duration was reduced by 14 ms at 60% repolarization (P < 0.0002) and stimulation threshold by 18% (P < 0.02). Epinephrine also allowed pacing at a cycle length of 140 ms, not achievable under control conditions. Because epinephrine decreases action potential duration at short cycle length in situ, re-entry wavefronts are less likely to encounter refractory tissue: fibrillation is more likely to occur and to remain stabilised. Reduction in action potential duration and excitation threshold were reversed by propranolol, suggesting that epinephrine effects are produced by beta-stimulation. PMID- 9201630 TI - The effect of alterations to action potential duration on beta-adrenoceptor mediated aftercontractions in human and guinea-pig ventricular myocytes. AB - Aftercontractions induced by beta-adrenoceptor stimulation in human and guinea pig cardiomyocytes may be related to changes in action potential duration (APD). We investigated the effects of altering APD during the occurrence of isoproterenol-induced aftercontractions, using the KATP channel openers cromakalim and lemakalim or the action potential voltage clamp technique, in guinea-pig and human ventricular cardiomyocytes. Contractile responses were measured at 32 degrees C using a video-based edge-detection system. In guinea-pig myocytes, action potentials, Indo-1 fluorescence and contraction were measured at 22 degrees C. Isoproterenol (< or = 12 nM) had variable effects on APD but induced aftercontractions, the majority (14/19 cells) of which occurred during the action potential. Short action potentials were produced using K+ channel openers. These compounds reduced or completely abolished the isoproterenol induced aftercontractions. Increasing isoproterenol in the presence of K+ channel opener restored the main contraction to a level similar to or above those with isoproterenol alone, but without the reappearance of aftercontractions. When cells were stimulated to contract under action potential voltage clamp, isoproterenol-induced aftercontractions were abolished by voltage clamping with action potentials of short duration. It was possible to induce aftercontractions in some cells without application of isoproterenol if voltage clamp-imposed action potentials of very long duration were used. These aftercontractions were also abolished by shortening action potential duration. We conclude that K+ channel openers or the imposition of action potentials of short duration can dissociate positively inotropic beta-adrenoceptor stimulation from aftercontraction formation and that action potentials of long duration can be pro arrhythmic. PMID- 9201631 TI - Sepsis alters myocardial and plasma concentrations of endothelin and nitric oxide in rats. AB - Cardiovascular derangements during sepsis may arise from a mismatch between endothelin (ET) and nitric oxide (NO). We hypothesized that progression of chronic peritoneal sepsis would affect cardiac performance and would modulate the concentrations of NO and ET in the heart and plasma. Male Sprague-Dawley rats (340-390 g) were catheterized and made septic with a cecal slurry (200 mg/kg: i.p.). Heart rate, mean arterial pressure, and plasma ET and nitrite/nitrate (NOX) were determined at 0, 4, 8, 12, 24, and 48 h after induction of sepsis. Septic rats were found to have tachycardia at 48 h following induction of sepsis. Mean arterial pressure and pulse pressure were not altered in septic and non septic rats. In a separate series of experiments, the function of isolated hearts from septic and non-septic rats was assessed at preload pressures of 2, 5, and 10 mmHg. Sepsis produced a significant decrease in rates of pressure development and relaxation (+/-dP/dt) at 24 and 48 h as compared to the hearts of non-septic rats. In septic rats, plasma concentrations of ET were significantly increased at t = 4, 8, 12 h as compared to basal values, and at 12 h as compared to non-septic rats, and returned to basal levels at 24 and 48 h. In contrast, circulating NO levels did not become elevated until t = 8 h and remained elevated throughout the remaining times. In the left ventricle, the concentration of ET was found to be significantly increased both in septic and non-septic rats at 4 and 8 h as compared to t = 0 h. In the left ventricles of non-septic rats, ET levels returned to baseline values at 12 h, while in septic rats, the concentration of ET remained significantly elevated until 12 h. In septic rats, left ventricular NO levels were found to be significantly increased at t = 12 h. It appeared that induction of sepsis contributed to an imbalance in the plasma concentration of ET and NO 12 h after the induction of sepsis. However, a similar imbalance was not observed in the left ventricle. It is concluded from these observations that peritoneal sepsis in a chronic rat model produced a divergence of plasma NO and ET levels. This suggests a homeostatic imbalance between vasoactive mediators, i.e. ET and NO, could contribute to the cardiovascular derangements that occur during sepsis. PMID- 9201632 TI - Receptor-independent activation of cardiac adenylyl cyclase by GDP and membrane associated nucleoside diphosphate kinase. A new cardiotonic mechanism? AB - Regulation of adenylyl cyclase activity by guanine nucleoside tri- and diphosphates as well as by stimulatory and inhibitory receptors was studied in canine cardiac sarcolemmal membranes. Guanosine triphosphate (GTP) increased adenylyl cyclase activity by a maximum of 80%, with an EC50 value of 0.7 mumol/l. The addition of the beta-adrenoceptor agonist, isoprenaline (100 mumol/l), caused a further, about 100%, increase in GTP-stimulated activity. The nucleoside diphosphate (GDP) also activated cardiac adenylyl cyclase, but in a biphasic manner. At low concentrations (EC50 0.12 mumol/ l). GDP increased enzyme activity by about 80%, followed by a plateau at 0.5-2 mumol/l and a second increase to a maximum of 60% with an EC50 value of 14 mumol/l. The stable GDP analog, guanosine 5'-O-(2-thio)diphosphate (GDP beta S), also increased cardiac adenylyl cyclase activity, but in a monophasic manner, by a maximum of 150%, with an EC50 of 0.4 mumol/l. Addition of uracil diphosphate (UDP) (3 mmol/l), which completely inhibited transphosphorylation of GDP to GTP, did not reduce adenylyl cyclase stimulation by low concentrations of GDP, whereas enzyme stimulation by high GDP concentrations was almost completely attenuated. Furthermore, pretreatment of the membranes with cholera toxin led to an increased stimulation of adenylyl cyclase activity by high concentrations of GDP. These findings suggest that the second phase of adenylyl cyclase stimulation by GDP is due to transphosphorylation of GDP to GTP, associated with activation of Gs proteins, and that stimulation by GDP itself (first phase) and endogenously formed GTP (second phase) is additive. However, in contrast to exogenously added GTP, beta-adrenoceptor activation did not enhance GDP-stimulated adenylyl cyclase activity. Furthermore, in the presence of 1 mumol/l GDP, the addition of GTP did not cause any further increase in enzyme activity. On the other hand, the muscarinic acetylcholine receptor agonist carbachol inhibited both GTP- and GDP-activated adenylyl cyclase. The inhibition of GDP-stimulated activity was lost when formation of GTP from GDP was blocked. The contrasting effects of endogenously formed GTP and exogenous GTP suggest that the formation of GTP from GDP is closely linked to the activation site of adenylyl cyclase, i.e. the stimulatory Gs protein. This receptor independent activation can apparently bypass beta-adrenoceptor-dependent activation of cardiac adenylyl cyclase. PMID- 9201633 TI - Regionally different vascular response to vasoactive substances in the remodelled infarcted rat heart; aberrant vasculature in the infarct scar. AB - Remodelling after myocardial infarction (MI) is associated with vascular adaption, increasing vascular capacity of non-infarcted myocardium, and angiogenesis in the infarcted part during wound healing and scarring. We investigated regional vascular reactivity in the infarcted rat heart. Transmural infarction of the left ventricular free wall was induced by coronary artery ligation. After 3 weeks, regional flow during maximal vasodilation (nitroprusside, NPR) and submaximal vasoconstriction (arginine-vasopressin, AVP) were studied in buffer-perfused hearts. The main findings were: (1) a reduced vasodilator response (NPR) in the viable part of the left ventricular free wall, where hypertrophy was most pronounced, resulting in reduced maximal tissue perfusion of the myocardium bordering the scar (19.7 + 0.6 v 25.7 + 1.2 ml/min.g), whereas perfusion of other non-infarcted regions was preserved. (2) A 54% lower vasodilator response (NPR) and a 25% stronger vasoconstriction (AVP) in scar tissue compared to viable parts of MI hearts. Microscopy showed thicker walls of resistance arteries in scar tissue than in viable parts of MI hearts or in sham hearts, morphometrically substantiated by two- to three-fold greater wall/lumen ratios. These data indicate a deviant response of scar vessels of MI hearts, and in the non-infarcted part, a reduced coronary reserve in the most hypertrophied region. Whereas the former may be caused by different vessel structure, the reduced vasodilator reserve of the spared part of the left ventricular free wall may indicate vasodilation at rest due to insufficient vascular growth. Thus, the most hypertrophied region would be at the highest risk of further ischemic damage. PMID- 9201634 TI - Direct gene transfer into the mouse heart. AB - Direct injection of plasmid DNA into the myocardium of several species has been shown to be useful for studying cardiac gene expression. However, despite a better understanding of mouse genetics and the availability of several disease models in mice, gene injection with plasmid DNA into the mouse heart has not been reported. In this study, we demonstrate a simple and reproducible method for gene transfer into the mouse heart via direct injection of plasmid DNA. A firefly luciferase gene, driven by the RSV promoter, was used to quantitatively determine the spatial and temporal characteristics of gene transfer. Luciferase gene expression was stable for 8 weeks and showed a dose-dependent response over a range of 0.3-3 micrograms of input DNA. Inter-animal variability was low and gene expression was restricted to the left ventricle, near the site of injection. This method was also demonstrated to be suitable for detecting the expression of structural genes under the control of cellular promoters. Immunohistochemistry was used to detect the expression of an epitope-tagged myosin heavy chain driven by a rat alpha-myosin heavy chain promoter. Thus, naked DNA injection into the mouse heart results in a highly reproducible expression of constructs with either viral or cellular promoters. It is a relatively inexpensive and efficient means of studying cardiac gene regulation in vivo and a useful tool for screening the potential transgenes before generating transgenic mice. PMID- 9201635 TI - Laxity of knee cruciate ligaments during pregnancy. AB - Pregnancy-related increase in ligament laxity may cause joint instability. The purpose of this study was two-fold: 1) to assess knee laxity changes during pregnancy and 2) to evaluate the effect of exercise on knee laxity due to a typical prenatal fitness program. The subjects were healthy pregnant women. One group (N = 27) participated in exercise classes designed according to national guidelines. The second group (N = 38) was more sedentary. A clinical arthrometer, KT-1000, was used, and anterior and posterior drawer tests were performed. The results were added and averaged for the two knees. Laxity was constant in the second half of pregnancy and had significantly decreased by 14% 4 months after birth. No influence of parity or exercise was detected. The exercise program employing minimal to moderate weight bearing did not result in any measurable increases in knee laxity and, therefore, appears to be appropriate with regard to knee stability. These results should not, however, be extended to different types of exercise programs without additional research. PMID- 9201636 TI - Comparison of nonballistic active knee extension in neural slump position and static stretch techniques on hamstring flexibility. AB - Nonballistic, active range of motion exercises have been advocated as more effective than static stretching for increasing range of motion, yet no published data exist to support this claim. This study compared the effect of nonballistic, repetitive active knee extension movements performed in a neural slump sitting position with static stretching technique on hamstring flexibility. Forty healthy, adult volunteer subjects with limited right hamstring flexibility (i.e., minimum of 15 degrees loss of active knee extension measured with femur held at 90 degrees of hip flexion) were randomly assigned to one of three groups. Group 1 (static stretch) performed a 30-second stretch twice daily. Group 2 (active stretch) performed 30 repetitions of active knee extension while sitting in a neural slump position twice daily. Group 3 served as a control. Hamstring flexibility was determined by an active knee extension test before and after 6 weeks of stretching. Goniometric measurement of knee joint flexion angle was obtained from videotape recording of the active knee extension test. A 3 (group) x 2 (test) repeated measures analysis of variance and subsequent Tukey post hoc testing revealed no significant difference in knee joint range of motion gains between the static (mean = 8.9 degrees) and active stretch (mean = 10.2 degrees). Both stretch groups' knee joint range of motion improved significantly (p < .05) more than the control group. We conclude that 6 weeks of nonballistic, repetitive active knee extensions (30 repetitions, twice daily) performed in a neural slump sitting position improves hamstring flexibility in uninjured subjects, but is no different compared with static stretching (30 seconds, twice daily). PMID- 9201637 TI - The efficacy of cryotherapy following arthroscopic knee surgery. AB - Cryotherapy has historically been used as a treatment following knee surgery. In the literature, there is little evidence of beneficial effects which support this practice. This study examined the effects of cryotherapy treatments on 45 subjects following minor arthroscopic knee surgery. Subjects were randomized to one of two treatment groups and the assessor remained blind to treatment group allocation. Subjects performed a 1-week home program of either cryotherapy and exercises or exercises alone. One week following surgery, a statistically significant difference was found between the groups for the affective dimension of the McGill pain questionnaire, medication consumption, compliance, and weight bearing status. No significant differences were found between the groups for other outcome variables. These results indicate that the addition of cryotherapy to a regime of exercises following arthroscopic knee surgery produced benefits of increased compliance, improved weight-bearing status, and lower prescription medication consumption. PMID- 9201638 TI - Preferential activation of the vastus medialis oblique, vastus lateralis, and hip adductor muscles during isometric exercises in females. AB - Disagreement exists as to whether the individual components of the quadriceps femoris can be preferentially activated, i.e., that one muscle component is activated to a greater degree of its maximum voluntary contraction ability than the remaining components. Preferential activation of the vastus medialis (VM) might be useful in the treatment of knee patients demonstrating VM atrophy. The purpose of the present investigation was to determine if the vastus medialis oblique (VMO), vastus lateralis (VL), and hip adductor (HA) muscles were preferentially activated in females during the following maximal voluntary isometric exercises: 1) unilateral quadriceps setting (QS) with the ankle positioned in neutral, 2) unilateral quadriceps setting combined with ankle dorsiflexion (QS + D), and 3) maximal bilateral hip adduction. Integrated electromyography (IEMG in mV.sec) was determined for the VMO, VL, and HA muscles of the preferred leg (i.e., that used to kick a ball) of 20 healthy females. Data were normalized using QS exercise as the reference exercise. Nonnormalized IEMG (+/-SD) of the VMO and VL was similar during QS [i.e., VMO = 1050 (+/-802) mV.sec, VL = 1075 (+/-738) mV. sec] and QS + D exercises [i.e., VMO = 1191 (+/ 738) mV.sec, VL = 1202 (+/-836) mV.sec], but significantly less than these values during hip adduction exercise [i.e., VMO = 174 (+/-62) mV. sec, VL = 194 (+/-70) mV.sec]. Nonnormalized IEMG of the HA muscles was similar during both QS and QS+D [i.e., 286 (+/-405) mV.sec and 195 (+/-432) mV.sec], but significantly higher than these values during hip adduction exercise [i.e., 413 (+/-235) mV.sec]. Normalized IEMG (+/-SD)(%) demonstrated similar patterns, i.e., the ratios for the VMO and the VL muscles did not differ from one another under either QS + D [i.e., VMO = 121 (+/-60)%, VL = 116 (+/-40)%] or hip adduction conditions [i.e., VMO = 33 (+/-24)%, VL = 36 (+/-25)%]. As a result, the degree of activation of the two muscles was considered the same. These results suggest no preferential activation of the quadriceps femoris component muscles during QS, QS + D, and hip adduction exercises in the nonweight-bearing position. The use of hip adduction to preferentially activate the VMO over the VL compared with QS exercises was not substantiated. A mean increase of 20% in the VMO and VL myoelectric activity during QS (as demonstrated by the normalized IEMG), by the addition of dorsiflexion, may be clinically significant. However, further study is required. PMID- 9201639 TI - The effects of "decelerated" rehabilitation following anterior cruciate ligament reconstruction on a hyperelastic female adolescent: a case study. AB - Current concepts in postoperative anterior cruciate ligament (ACL) reconstruction management include participation in an "accelerated" rehabilitation program. There are no published reports examining the effects of accelerated or conservative rehabilitation on subjects with generalized ligamentous hyperelasticity. The purpose of this case study was to examine the effects of a conservative or "decelerated" rehabilitation program on the functional outcome of a hyperelastic female adolescent athlete following ACL reconstruction. The subject was a 15-year-old female basketball player who sustained a unilateral ACL tear and underwent subsequent ACL reconstruction using a patellar tendon autograft. The subject immediately began participation in a "decelerated" rehabilitation program in which the intensity and rate of progression was decelerated, emphasizing a prolonged period of maximum graft protection. Progress was objectively quantified with a battery of diagnosis-specific tests at scheduled intervals. Results at 52 weeks postoperative revealed normal range of motion, proprioception, balance, knee stability, quadriceps strength, hamstring strength, and subjective assessment values, and only a 4.0% deficit in functional scores. Our results suggest a "decelerated" rehabilitation program may be appropriate for the population with generalized ligamentous hyperelasticity by yielding excellent functional results without compromising the integrity of the graft and, ultimately, knee stability. PMID- 9201640 TI - Case study: physical therapy management of hip osteoarthritis prior to total hip arthroplasty. AB - It is important that we have information on the role of physical therapists in the treatment of patients with osteoarthritis of the hip prior to total hip arthroplasty. This article describes the management of a patient with limited range of motion of the right hip due to osteoarthritis. The patient made a significant improvement with decreased pain, increased range of motion of the right hip, increased periarticular muscle strength, improved gait, and improved mobility. One year later, the patient had a right total hip arthroplasty. The rationale of the management of patients with osteoarthritis of the hip is discussed. In addition, the role of physical therapists in the management and treatment of patients with osteoarthritis prior to total hip arthroplasty is discussed. PMID- 9201641 TI - Follow-up to the clinical and cost-effectiveness of two different programs for rehabilitation following ACL reconstruction. PMID- 9201642 TI - Follow-up to the clinical and cost-effectiveness of two different programs for rehabilitation following ACL reconstruction. PMID- 9201643 TI - Epidemiologic advances in chronic fatigue syndrome. AB - Epidemiologic studies of chronic fatigue syndrome (CFS) have been hampered by the absence of a specific diagnostic test, but with increasing interest in this disorder there has been a greater understanding of the risk factors, illness patterns, and other aspects of this multisystem disorder. Working case definitions have been developed for research purposes but they have continued to change over time and have not always been utilized precisely by various investigators. This has been a major factor in the widely varying estimates of prevalence rates, but two different studies using the same working definition and including a medical work-up have estimated the prevalence to be approximately 200/100,000. Clusters of CFS cases, which appear to be related to earlier reports of "epidemic neuromyasthenia", have attracted considerable attention and appear to be well documented, although investigated with varying methodology and often with dissimilar case definitions. Risk factors for cases occurring in clusters and sporadically appear to be similar, the most consistent ones being female gender and the co-existence of some form of stress, either physical or psychological. The prognosis of CFS is difficult to predict, although cases occurring as part of clusters appear to have a better prognosis as a group than sporadic cases, and those with an acute onset have a better prognosis than those with gradual onset. It is highly unlikely that there is a single agent, infectious or noninfectious, that is responsible for more than a small proportion of CFS cases and, at the present time, the risk factors for developing CFS appear to lie more prominently in the host rather than the environment. PMID- 9201644 TI - An epidemiologic study of fatigue with relevance for the chronic fatigue syndrome. AB - We surveyed households in four rural Michigan communities to confirm a reported cluster of cases resembling chronic fatigue syndrome (CFS) and to study the epidemiology of fatigue in a rural area. Data were collected from 1698 households. We did not confirm the reported cluster. The prevalence of households containing at least one fatigued person was similar between communities thought to harbor the cluster and communities selected for comparison. Symptoms and features of generic forms of fatigue were very similar to those often attributed to CFS. PMID- 9201645 TI - Fatiguing illness among employees in three large state office buildings, California, 1993: was there an outbreak? AB - The objective was to determine if a cluster of chronic fatigue syndrome (CFS) like illness had occurred among employees in two large state office buildings in northern California, and to identify risk factors for and features of fatiguing illness in this population. DESIGN: case-control study. POPULATION AND SETTING: Over 3300 current employees in two state office buildings and employees in a comparable "control" building. Information was collected on demographic and occupational variables, the occurrence of fatiguing illness for at least one month in the previous year, and the presence of 36 symptoms. A total of 3312 (82%) of 4035 employees returned questionnaires. Overall, 618 (18.7%) persons reported fatigue lasting at least one month; including 382 (11.5%) with fatigue of at least six months' duration and 75 (2.3%) with symptoms compatible with a CFS-like illness. Independent risk factors for fatigue lasting one month or longer were found to be Native American ethnicity (OR 2.4, CI 1.1,5.3), Hispanic ethnicity (OR 1.7, CI 1.3,2.3), female sex (OR 1.5, CI 1.2,1.9), gross household incomes of less than $50,000 (OR 1.3, CI 1.1,1.6), and less than a college education (OR 1.3, CI 1.1,1.6). Similar risks were observed for persons who reported fatigue lasting six months or longer. Female sex (OR 3.2, CI 1.7, 6.4) was the only independent risk factor found for those persons classified as having a CFS-like illness. Case prevalence rates for all three categories of fatigue, as determined by multivariate analysis, were not significantly different among buildings. Despite finding a substantial number of employees with fatiguing illness in the two state office buildings, the prevalence was not significantly different than that for a comparable control building. Previously unidentified risk factors for fatigue of at least one month and at least six months identified in this population included Hispanic ethnicity, not having completed college, and income below $50,000. PMID- 9201646 TI - Chronic fatigue syndrome criteria in patients with other forms of unexplained chronic fatigue. AB - To determine the prevalence of chronic fatigue syndrome (CFS) criteria in other forms of unexplained chronic fatigue, 297 consecutive outpatients under the age of 40 from a general medicine practice were studied. After excluding the three with chronic fatigue syndrome, the remaining 294 individuals were divided into those with unexplained chronic fatigue (64 patients) those without (the remaining 230 patients). Chronic fatigue syndrome criteria noted to be significantly more common in those with unexplained fatigue compared to those without include: fever, painful adenopathy, muscle weakness, myalgia, headache, migratory arthralgia, neuropsychologic symptoms, and sleep disorder. Like chronic fatigue syndrome, unexplained chronic fatigue often started suddenly. I conclude that the CFS criteria are noted more commonly than expected in other forms of unexplained chronic fatigue. PMID- 9201647 TI - The natural history of concurrent sick building syndrome and chronic fatigue syndrome. AB - An outbreak of chronic fatigue syndrome linked with sick building syndrome was recently described as a new association. Whether chronic fatigue syndrome acquired in this setting tends to remit or, as sporadic cases often do, persist, is unknown. To clarify the natural history of chronic fatigue syndrome in association with sick building syndrome the 23 individuals involved in the outbreak were interviewed four years after the onset. In the previous interview one year after the onset of symptoms, 15 (including 5 with chronic fatigue syndrome and 10 with idiopathic chronic fatigue) of the 23 noted fatigue. Three years later 10 of the 15 were "fatigue free" or "much improved". Five were only "some better", "the same", or "worse". Three of the five people previously diagnosed with chronic fatigue syndrome were "much improved" (two) or "fatigue free" (one). The remaining two were seriously impaired, homebound and unable to work. The 10 individuals with substantially improved fatigue (three of the five with chronic fatigue syndrome and seven of the 10 with idiopathic chronic fatigue) were more likely to have noted improvement in nasal and sinus symptoms, sore throats, headaches, and tender cervical lymph nodes when compared to those with a lingering significant fatigue (p < 0.001). Upper respiratory symptoms and headaches improved in those with reduced fatigue but remained problematic in those with persisting significant fatigue. We conclude that the fatigue related to sick building syndrome, including chronic fatigue syndrome, is significantly more likely to improve than fatigue identified in sporadic cases of chronic fatigue syndrome. PMID- 9201648 TI - Precipitating factors for the chronic fatigue syndrome. AB - The etiology of the Chronic Fatigue Syndrome (CFS) is unknown but it is usually considered to be postinfectious or postviral. Many infecting agents have been suspected as causative but none has been proven. We investigated precipitating factors in 134 CFS patients through the use of a questionnaire, interview, clinical examination and serology for infecting agents; 35 healthy controls completed a similar questionnaire. CFS started with an apparently infectious illness in 96 (72%) but a definite infection was only found in seven of these 96 (7%). Thirty-eight (28%) had no apparent infectious onset: 15/38 (40%) had noninfectious precipitants (trauma, allergy, surgery). There was no apparent precipitating event in 23/38 (61%). Immunization was not a significant precipitant. Stressful events were very common in the year preceding the onset of CFS (114/134, 85%) but these occurred in only 2/35 (6%) of the controls (p < .0001). The onset of CFS may be associated with preceding stressful events and multiple other precipitants. An infectious illness is not uniformly present at the onset and no single infectious agent has been found; CFS is most likely multifactorial in origin. PMID- 9201649 TI - Neuroendocrine correlates of chronic fatigue syndrome: a brief review. AB - Chronic fatigue syndrome remains one of the more perplexing syndromes in contemporary clinical medicine. One approach to understanding this condition has been to acknowledge its similarities to other disorders of clearer pathophysiology. In this review, a rationale for the study of neuroendocrine correlates of chronic fatigue syndrome is presented, based in part on the clinical observation that asthenic or fatigue states share many of the somatic symptom characteristics seen in recognized endocrine disorders. Of additional interest is the observation that psychological symptoms, particularly disturbances in mood and anxiety, are equally prominent in this condition. At this time, several reports have provided replicated evidence of disruptions in the integrity of the hypothalamic-pituitary-adrenal axis in patients with chronic fatigue syndrome. It is notable that the pattern of the alteration in the stress response apparatus is not reminiscent of the well-understood hypercortisolism of melancholic depression but, rather, suggests a sustained inactivation od central nervous system components of this system. Recent work also implicates alterations in central serotonergic tone in the overall pathophysiology of this finding. The implications of these observations are far from clear, but they highlight the fact that, though chronic fatigue syndrome overlaps with the well-described illness category of major depression, these are not identical clinical conditions. PMID- 9201650 TI - Sudden vs gradual onset of chronic fatigue syndrome differentiates individuals on cognitive and psychiatric measures. AB - To examine the influence of mode of illness onset on psychiatric status and neuropsychological performance, 36 patients with CFS were divided into two groups: sudden vs gradual onset of symptoms. These two CFS subgroups were compared to each other and to sedentary healthy controls on standardized neuropsychological tests of attention/concentration, information processing efficiency, memory, and higher cortical functions. In addition, the distribution of comorbid Axis I psychiatric disease between the two CFS groups was examined. The rate of concurrent psychiatric disease was significantly greater in the CFS gradual group relative to the CFS-sudden group. While both CFS groups showed a significant reduction in information processing ability relative to controls, impairment in memory was more severe in the CFS-sudden group. Because of the significant heterogeneity of the CFS population, the need for subgroup analysis is discussed. PMID- 9201651 TI - Somatomedin C (insulin-like growth factor I) levels in patients with chronic fatigue syndrome. AB - Chronic fatigue syndrome is a disorder clinically quite similar to fibromyalgia syndrome, and it is of interest to examine if these two syndromes have pathogenetic as well as clinical features in common. Somatomedin C levels have been found to be lower in patients with fibromyalgia syndrome than in healthy controls. An attractive hypothesis relating sleep disturbance, altered somatotropic neuroendocrine function and fibromyalgia symptoms has been put forward as a plausible pathogenic mechanism for fibromyalgia syndrome. We therefore sought to investigate the level of somatomedin C in patients with chronic fatigue syndrome. Somatomedin C levels were determined by radioimmunoassay in frozen serum specimens from 49 patients with CFS and 30 healthy blood donor control subjects of similar age and gender. Somatomedin C levels were higher in patients with CFS than in healthy control subjects (255.3 +/- 68.5 vs 211.9 +/- 76.2, P = 0.01). There was no effect of gender, use of nonsteroidal anti-inflammatory drugs or tricyclic drugs on levels of somatomedin C. There was a tendency for somatomedin C levels to fall with age. In contrast to patients with fibromyalgia, in whom levels of somatomedin C have been found to be reduced, levels in patients with CFS were found to be elevated. Thus, despite the clinical similarities between these two conditions, they may be associated with different abnormalities of sleep and/or of the somatotropic neuroendocrine axis. PMID- 9201652 TI - Gender differences in host defense mechanisms. AB - Extensive studies in both humans and animals have shown that females express enhanced levels of immunoreactivity compared to males. Whereas this provides females with increased resistance to many types of infection, it also makes them more susceptible to autoimmune diseases. This review will focus on gender-related differences in non-specific host defense mechanisms with a particular emphasis on monocyte/macrophage function and a primary product of monocytes: interleukin-1 (IL-1). Immunomodulatory cytokines such as IL-1 are influenced by gender sensitive hormones, and reciprocally, these cytokines influence gender-specific hormones and tissues. Patients with chronic fatigue syndrome (CFS) are predominantly women, therefore it may be useful to look toward gender-specific differences in immune function to find a key for this poorly understood syndrome. PMID- 9201653 TI - Electron microscopic immunocytological profiles in chronic fatigue syndrome. AB - Structures consistent in size, shape and character with various stages of a Lentivirus replicative cycle were observed by electron microscopy in 12-day peripheral-blood lymphocyte cultures from 10 of 17 Chronic Fatigue Syndrome patients and not in controls. Attempts to identify a lymphoid phenotype containing these structures by immunogold labelling failed and the results of reverse-transcriptase assay of culture supernatants were equivocal. The study was blind and case-controlled, patients being paired with age, sex and ethnically matched healthy volunteers. Prescreening of subjects included the common metabolic and immunological disorders, functional conditions and a virus-screen against hepatitis B and C, Epstein-Barr Virus, Cytomegalovirus and Human Immunodeficiency Virus. PMID- 9201654 TI - The relationship between fibromyalgia and interstitial cystitis. AB - Interstitial cystitis (IC) is a relatively uncommon and enigmatic disorder characterized by pain in the bladder and pelvic region, typically accompanied by urinary urgency and frequency. Fibromyalgia is a more common disorder, with the prominent symptoms being diffuse musculoskeletal pain and fatigue, and it has been well established that there is substantial clinical overlap between fibromyalgia and chronic fatigue syndrome (CFS). Although genitourinary and musculoskeletal symptoms predominate in IC and fibromyalgia respectively, both disorders share a number of features, including similar demographics, "allied conditions" (e.g. irritable bowel syndrome, headaches, etc.), natural history, aggravating factors, and efficacious therapy. We hypothesized that there was substantial clinical overlap between fibromyalgia and IC, and examined cohorts of individuals with these two disorders in parallel, to compare the spectrum of symptomatology. Sixty fibromyalgia patients, 30 IC patients, and 30 age-matched healthy controls were questioned regarding current symptomatology. A dolorimeter examination was also performed in the three groups to assess peripheral nociception. We found that the frequency of current symptoms was very similar for the fibromyalgia and IC groups. Both the fibromyalgia and IC patients displayed increased pain sensitivity when compared to healthy individuals, at both tender and control points. These data suggest that IC and fibromyalgia have significant overlap in symptomatology, and that IC patients display diffusely increased peripheral nociception, as is seen in fibromyalgia. Although central mechanisms have been suspected to contribute to the pathogenesis of fibromyalgia for some time, we speculate that these same types of mechanisms may be operative in IC, which has traditionally been felt to be a bladder disorder. PMID- 9201655 TI - Double-blind randomized controlled trial to assess the efficacy of intravenous gammaglobulin for the management of chronic fatigue syndrome in adolescents. AB - A double blind randomized controlled trial was conducted in 71 adolescents aged 11-18 years. Inclusion in the trial required fulfilment of the diagnostic criteria, (Fukuda et al., 1994). Three infusions of 1 gm/kg (max 1 litre of 6 gm/100 ml in 10% w/v maltose solution) were given one month apart. The dummy solution was a 10% w/v maltose solution with 1% albumin of equivalent volume for weight. Efficacy was assessed by difference in a mean functional score including school attendance, school work, social activity and physical activity, between baseline, three months and six months after the final infusion. There was a significant mean functional improvement at the six month follow-up of 70 adolescents with Chronic Fatigue Syndrome of average duration 18 months. There was also a significant improvement for both groups from the beginning of the trial to the six month post infusion follow-up. Adverse effects were common with both solutions but not predictive of response. Neither solution could be identified by recipients. PMID- 9201656 TI - Cytokine production by adherent and non-adherent mononuclear cells in chronic fatigue syndrome. AB - It has been suggested that cytokines play a role in certain clinical manifestations of chronic fatigue syndrome (CFS). In this study adherent (monocytes) and non-adherent (lymphocytes) mononuclear cells were stimulated in the presence or absence of phytohemagglutinin (PHA) or lipopolysaccharide (LPS), respectively, and supernatants were assayed for IL-6, TNF-alpha, and IL-10 by ELISA. IL-6 was also measured at the mRNA level by polymerase chain reaction. The levels of spontaneously (unstimulated) produced TNF-alpha by non-adherent lymphocytes and spontaneously produced IL-6 by both adherent monocytes and non adherent lymphocytes were significantly increased as compared to simultaneously studied matched controls. The abnormality of IL-6 was also observed at mRNA level. In contrast, spontaneously produced IL-10 by both adherent and non adherent cells and by PHA-activated non-adherent cells were decreased. This preliminary study suggests that an aberrant production of cytokines in CFS may play a role in the pathogenesis and in some of the clinical manifestations of CFS. PMID- 9201657 TI - The tobacco dilemma: from warning to banning--or just plain smoke? PMID- 9201658 TI - The marvelous egg. PMID- 9201659 TI - After the menopause: tamoxifen and other new prevention maintenance therapies. PMID- 9201660 TI - Are observational data adequate to guide lipid altering therapy in women? AB - The epidemiologic data presented by Emond and Zareba in this issue provide interesting observations of the relationships among cholesterol concentrations, age, and risk of CHD events and death. The data they present did not incorporate some important variables in the CHD risk profile for women. The absence of good information on diabetes mellitus, menopause, and HRT and of adequate HDL-C data means that these data cannot be used to guide lipid-altering treatment. Other observational datasets and limited current intervention trial data suggest that women with documented CHD and women who, in the absence of CHD, have high risk for CHD should be treated according to currently recommended (i.e., NCEP) guidelines. Treatment of lipid levels in very low-risk patients is less clear. Data from clinical studies currently under way will define the potential benefits of lipid-altering therapy. Intervention trials with HRT will likely give us insight about the exact role of this modality in CHD risk reduction in women. Results of these trials should be available within the next decade. PMID- 9201661 TI - Physician as detective: discerning and recording domestic violence injuries. PMID- 9201662 TI - Observations from the CDC. The Prevention Research Centers Program: collaboration in women's health. PMID- 9201663 TI - Long-term follow-up of gender-specific outcomes after thrombolytic therapy for acute myocardial infarction from the GUSTO-I trial. Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries. AB - Our objective was to assess gender differences in mortality 1 year after acute myocardial infarction (MI). The Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries (GUSTO-I) trial database of 41,021 patients with suspected acute MI was used to generate 1-year Kaplan-Meier survival plots. Risk quartiles and mortality of women and men were compared. The unadjusted 1-year mortality rate for the initial GUSTO-I population and 30-day survivors demonstrates a large gender gap [odds ratio for all patients = 2.2, 95% confidence interval (CI), 2.0-2.3, p < 0.001]. For the initial population, when adjusted for age, the gender gap is still apparent (odds ratio = 1.4, 95%, CI = 1.3-1.5, p < 0.001) although no longer significant when adjusted using the 30-day survival model (odds ratio = 1.06, 95% CI = 0.97-1.15, p < 0.001). For the 30-day survivors, adjustment based on age alone explained the 1-year mortality difference (risk ratio = 0.96, 95% CI 0.85-1.07, p = 0.441). When the population was divided into expected risk quartiles, women were more likely to fall into the higher expected risk quartiles, even after adjusting for age. A gender gap after acute MI is apparent, nearly all of which occurs within the first 30 days. A substantial portion of the gender gap is explained by the increased age of women, and the rest of the gap may be attributed to differences in variables predictive of 30-day mortality. During 1-year follow-up, the late mortality of women is no greater than that of age-matched men. PMID- 9201664 TI - Prognostic value of cholesterol in women of different ages. AB - We assessed the short-term and long-term prognostic relationship between cholesterol and mortality in women of different ages with the aid of statistical graphics. Our population-based cohort study involved 2873 women in the Framingham Heart Study, with a median follow-up of 31 years. The primary outcome was all cause mortality. Secondary outcome measures were coronary heart disease, noncoronary heart disease, and stroke mortality. We found that significant age interactions were present in the relationships between total cholesterol and mortality from all causes, coronary heart disease (CHD), stroke, and non-CHD causes. For women ages < or = 55, cholesterol is related positively to both short term (p > 0.05) and long-term (p = 0.05) all-cause mortality. For women ages 56 70, there are significant U-shaped relationships between cholesterol and both short-term and long-term all-cause mortality (p < 0.01). Lowest short-term and long-term mortality rates for women in this age group are at cholesterol values between 240 and 280 mg/dl. For women ages > 70, cholesterol < 240 mg/dl is associated with increased short-term mortality (p < 0.01), and no significant long-term association was detected. These cholesterol/mortality relationships and age interactions can be explained by patterns of association between mortality and both high- and low-density lipoprotein cholesterol among women in the different age groups. These results do not support the hypothesis that cholesterol < 200 mg/dl leads to decreased mortality in women > 55 years old. PMID- 9201665 TI - Epidemiology of hysterectomy in the United States: demographic and reproductive factors in a nationally representative sample. AB - We describe the epidemiology of hysterectomy, overall as well as for specific indications. Data were obtained from the Epidemiologic Follow-up to the First National Health and Nutrition Examination Survey, a nationally representative cohort followed prospectively from the mid-1970s through 1992. Black and white women 25-49 years of age, interviewed during follow-up, were included in the analyses. The probability of undergoing a hysterectomy was estimated by demographic and reproductive factors. Hysterectomy as confirmed by hospital records was our main outcome measure. We found that women who had completed 9-11 years of education were more likely to have undergone a hysterectomy than were women with either more or less education. Women who had completed 9-11 years of education were also more likely to have had a hysterectomy because of menstrual problems. Three or more miscarriages, especially if caused by uterine prolapse, increased the probability of hysterectomy. Having had no live births decreased the probability of hysterectomy for menstrual disorders and uterine prolapse, but women who had their first child before age 20 were at increased risk of hysterectomy because of endometriosis. Hysterectomy appears to be associated with low education, high parity, and a history of multiple miscarriages. The influence of these factors varies depending on the primary indication for the hysterectomy. PMID- 9201666 TI - Genetic testing for susceptibility to breast cancer: findings from women's focus groups. AB - Before designing an intervention to assist women in making informed decisions about BRCA1 testing, we conducted focus groups with women who had breast cancer and unaffected women whose relatives had it to better understand women's knowledge, concerns about testing, and potential influences and support needs in making a decision about genetic testing for susceptibility to breast cancer. Findings show a general lack of knowledge about genetic testing for breast cancer and what it means to have a positive test result, a strong concern for family members, particularly daughters, to use information from testing to help them make better decisions about their health and lifestyle choices, a strong sense of altruism, particularly among affected women, about being tested to help other women, not just family, and various support needs surrounding the testing experience, including an active role for physicians in the decision process. The major advantages to testing seem to be for information that could help reduce uncertainty and assist with making future decisions about medical treatment and plans for surveillance and some lifestyle changes. The major disadvantages to testing were concerns about confidentiality and loss of insurance, the lack of proven options for women after testing, and stress from knowing one had the BRCA1 mutation. These focus group discussions show women's concerns and ambivalence about genetic testing. We need to provide women with balanced information about the positive and negative aspects of such testing, determine how best to involve physicians in women's decisions about testing, consider the effects of testing on family relationships, and provide more public education about what genetic testing is and what it means. PMID- 9201667 TI - Women's health as a paradigm for understanding factors that mediate disease. AB - Women's health research provides an opportunity to focus on the mechanisms underlying health and disease in both men and women. We propose that differences in risk between men and women and among individual women are primarily determined by biologic factors, such as sex steroid hormone metabolism, anatomy, immunologic function, genetic influences, and the effects of reproduction, interacting with external influences, such as psychologic development, sociocultural environment, and economic status. We provide examples whereby a comparison of disease frequencies between men and women and among women allows a focus on underlying mechanisms of disease. We then demonstrate the importance of posing questions about the genesis of differences in disease frequency. PMID- 9201668 TI - Fluoxetine vs. tricyclic antidepressants in women with major depressive disorder. AB - Major depressive disorder and dysthymia are twice as prevalent in women as in men, and the lifetime risk of a woman's developing a major depressive disorder is about 20%. Yet depression is often unrecognized or misdiagnosed in women, and only about one quarter of women who meet criteria for major depressive disorder receive appropriate therapy. Until recently, women were generally excluded from clinical drug trials because of concerns of inadvertent pregnancy and risk of teratogenicity. Thus, information on safety and efficacy in those most likely to require antidepressant therapy is lacking. Studies have shown that the antidepressant fluoxetine, a selective serotonin reuptake inhibitor (SSRI) has a more tolerable side effect profile than do tricyclic amine (TCA) antidepressants, but few data have been reported on the efficacy and tolerability of fluoxetine or other SSRIs in female patients. In this study, a retrospective analysis of 11 randomized, double-blind, well-controlled trials was done to compare data from 427 female patients on fluoxetine and 423 female patients on TCAs. Both fluoxetine and TCAs significantly reduced the HAMD17 total mean score from baseline to end point, week 5 (fluoxetine, 24.35 to 14.37; TCAs, 24.57 to 14.43; p < 0.001). Both treatment groups were associated with significant reductions in the HAMD17 anxiety/somatization and insomnia subfactor scores. Abnormal vision, constipation, dizziness, dry mouth, and somnolence occurred more frequently (p < 0.05) in the TCA group. Insomnia and nausea were the only adverse events more common (p < 0.05) in the fluoxetine group. This study demonstrates that fluoxetine is an effective and tolerable agent for the treatment of major depressive disorder in women. PMID- 9201669 TI - Search engines. PMID- 9201670 TI - Validation of family history of breast cancer and identification of the BRCA1 and other syndromes using a population-based cancer registry. PMID- 9201671 TI - Chorionic tumours. PMID- 9201672 TI - Reproductive health LiteratureWatch. PMID- 9201673 TI - T2-weighted MR imaging of the liver: optimization of hybrid-RARE sequences. AB - The objectives of this study were to optimize T2-weighted hybrid-RARE pulse sequences for clinical MR imaging of the liver, and to compare them to the conventional spin-echo (CSE) sequence. Specifically, the ranges of the echo train length (ETL) and the effective echo time (TEeff) were investigated to optimize image quality and liver-spleen contrast, in healthy volunteers. A total of thirteen volunteers were scanned at 1.5 Tesla with an extensive array of hybrid RARE scans. The images were analyzed quantitatively with respect to CNR (contrast to-noise ratio of spleen vs. liver), SNR (signal to noise ratio of the spleen), SIR (signal intensity ratio of liver and spleen) and CDR (contrast difference ratio between the spleen and liver). The images were also analyzed qualitatively with respect to image sharpness, vascular artifacts, ghosting, chemical shift, and truncations. Results of quantitative analysis indicated that CDR and SIR of hybrid-RARE at higher ETL (> 13) were consistently better than both the reference CSE and the lower ETL sequences (p < 0.05) at all TEeff. SNR was slightly inferior for all hybrid-RARE sequences than for the CSE sequence. Image quality for hybrid-RARE sequences with ETL > 13 proved to be consistently better than that for the CSE (TE = 90 ms) with respect to imaging sharpness, vascular artifacts and ghosting artifacts (p < 0.05). In conclusion, the optimized hybrid RARE sequences with ETL greater than or equal to 13 are capable of producing sharp and relatively artifact free images with the advantage of a much greater acquisition time efficiency. PMID- 9201674 TI - Characterization of focal hepatic masses by dynamic contrast-enhanced MR imaging: findings in 311 lesions. AB - This study aimed to determine the overall accuracy of known enhancement patterns for the characterization of a large series of focal hepatic masses on dynamic contrast-enhanced magnetic resonance (MR) images. Breath-hold T1-weighted images of the liver acquired before intravenous gadolinium administration and serially over 6-10 min after contrast injection were acquired in < 25 a using FLASH or rapid spin-echo pulse sequences. A total of 311 proven focal hepatic masses in 128 patients were analyzed, including 192 malignant lesions (166 metastases and 26 hepatomas) and 119 benign lesions (48 cavernous hemangiomas, 45 hepatic cysts, and 26 other abnormalities). The lesions were evaluated for a variety of characteristics independently by two reviewers who were blinded to results. Cavernous hemangiomas showed early peripheral nodular enhancement (80% sensitivity and 99% specificity) and hepatic cysts showed no enhancement (100% sensitivity and 95% specificity). Hepatic metastases showed variable, moderate enhancement (47% by one reviewer and 74% by the other). Metastatic lesions from hypervascular primary neoplasms displayed peak enhancement during the hepatic artery dominant (bolus) phase, while other malignant neoplasms showed later peak enhancement (72% sensitivity and 77% specificity). Five metastatic foci with early homogeneous enhancement showed a delayed peripheral washout of contrast (rim sign), while no nonmetastatic foci displayed this finding (3% sensitivity and 100% specificity). Characteristic enhancement patterns of focal hepatic lesions were described in a large series of patients. This study confirms results of previous investigators who have shown that early nodular peripheral enhancement was highly specific for hemangiomas and lack of enhancement was highly specific for hepatic cysts. Hypervascular metastatic foci show earlier peak enhancement than other malignant lesions. Some (2-3%) metastatic lesions display a peripheral washout of contrast on serial images, with 100% specificity. PMID- 9201675 TI - Introduction of fast MR imaging in the assessment of hepatic steatosis. AB - We determined the utility of fast gradient echo techniques (modified Dixon method) in the assessment of hepatic fat content. Fast spoiled gradient echo was performed on bovine liver/corn oil homogenates with known fat fractions (FFE) to assess the accuracy of fat quantitation (FFMRI). The pulse sequence was manipulated via alterations in TE (echo time), TR (repetition time), and alpha (flip angle). In vivo studies were then performed using breath-holding maneuvers on normal adult volunteers and subjects at risk to develop hepatic steatosis, with cystic fibrosis or morbid obesity. At out-of-phase, TE, TR, and alpha were 2.1 ms, 7.3 ms, and 30-50 degrees and in-phase TE, TR, and alpha were 4.2 ms, 9.3 ms, and 30-50 degrees; FFMRI correlated well with FFE. An elevated fat fraction was observed in a high percentage of subjects with cystic fibrosis and morbid obesity. Fast gradient echo techniques were used successfully in the assessment of hepatic steatosis. The reduced acquisition times permitted in vivo analysis on adults and children using breath hold maneuvers. PMID- 9201676 TI - Dynamic multi-planar EPI of the urinary bladder during voiding with simultaneous detrusor pressure measurement. AB - Magnetic resonance imaging gives high quality images of the urinary bladder with excellent contrast. We report here the first application of dynamic, multi-slice, echo planar imaging to a study of urinary bladder emptying. Changes in urinary bladder volumes and rates of urine expulsion from the bladder have been measured simultaneously with bladder pressure. The method shows promise for clinical applications involving compromised bladder function, for reappraising bladder contraction strength-volume relationships, and for investigating the rate of change of length, three-dimensional shape, and wall tension in different parts of the bladder during micturition. PMID- 9201677 TI - A study of the effects of patient anxiety, perceptions and equipment on motion artifacts in magnetic resonance imaging. AB - We investigated to see if motion artifacts (MA) occurring in magnetic resonance imaging (MRI) are related to prescan anxiety measures and test the feasibility of identifying patients at risk for the development of MA before scanning. Furthermore, to determine a possible influence of constructional differences between a 1.5 and a 0.5 tesla scanner on the frequency of MA. Two hundred and ninety-seven first time MRI patients were surveyed before and after imaging with anxiety and attitude questionnaires. Frequency and impact on diagnostic quality of MA were documented. 12.8% of all scans showed MA not related to normal body pulsations. In 6.4% the diagnostic quality was impaired. Constructional differences did not influence the frequency of MA. Also, anxiety as determined with the most common anxiety measuring instrument was not related to the development of MA. Concern about the technical apparatus identified 70.6% of all individuals developing MA. Patients at risk for the development of MA can be identified prior to scanning. It seems necessary to further develop reliable methods to detect them and to evaluate strategies to prevent MA. PMID- 9201678 TI - Accuracy of gamma-variate fits to concentration-time curves from dynamic susceptibility-contrast enhanced MRI: influence of time resolution, maximal signal drop and signal-to-noise. AB - Concentration-time curves derived from dynamic susceptibility-contrast enhanced magnetic resonance imaging are widely used to calculate cerebrovascular parameters. To exclude effects of recirculation, a non-linear regression method is used to fit a gamma-variate function to the concentration-time course. In previous studies the errors arising from the fitting procedure have not been quantified. In a computer simulation we investigate the uncertainties of parameters calculated from the fitted gamma-variate function, exploring the dependencies on signal-to-noise (SNR), time resolution (delta t), and maximal signal drop (MSD). Our study was performed to give a framework on how to design MR-sequences and choose contrast media and their application in order to yield concentration-time curves which allow a reliable performance of the gamma-variate fitting procedure. We recorded 396 concentration-time curves from regions of interest of 40 patients. The gamma-variate fitting procedure was applied to these curves resulting in 396 parameter sets. Ideal concentration-time curves as gamma variate functions were generated from these sets with a given delta t, MSD, and SNR. Recirculation effect was simulated. Then the gamma-variate fitting was performed again. From ideal and simulated gamma-variate function the area and the normalized first moment were calculated. The uncertainties of the values calculated from the simulated curve relating to the values of the original one were determined. Increase of SNR decreases the involved errors. With SNR values of 100 and more there is only minor influence of delta t and MSD and the fitted curve approximates the original data very well. Smaller values of SNR lead to a stronger influence of delta t and MSD and a higher number of fitting failures. With increasing delta t the uncertainties also increase. Intermediate values of MSD (30% to 70%) yield the smallest errors while increasing or decreasing MSD yields an increase of uncertainty. To achieve low uncertainties in the calculation of cerebrovascular parameters from gamma-variate fits, delta t of the imaging sequence and MSD must be considered. This is more important the lower SNR is. The shown dependencies should be taken into account when choosing MR sequence parameters and application of contrast media. PMID- 9201679 TI - The use of active noise control (ANC) to reduce acoustic noise generated during MRI scanning: some initial results. AB - MRI scanning generates high levels of acoustic noise that cannot only pose a safety hazard, but also impair communication between staff and patient. In this article we present active noise control (ANC) techniques that introduce antiphase noise to destructively interfere with the MRI noise and with the aim of producing a zone of quiet around the patient's ears. Using noise recorded from a 1.0 Tesla midfield MR scanner the acoustic noise generated by three standard MR imaging sequences was replayed to a real time two channel ANC system. The results obtained show a useful attenuation of low-frequency periodic acoustic noise components. Therefore, in combination with standard passive ear protection, this suggests that MR generated acoustic noise can be effectively attenuated at both low and high frequencies leading to improved patient comfort. PMID- 9201681 TI - T2 relaxation times of irradiated vertebral bone marrow in patients with seminoma. AB - Our purpose was to demonstrate the effects of localized radiotherapy on lumbar vertebral bone marrow with the use of quantitative MRI with measurements of T2 relaxation times. Ten patients with early stage testicular seminoma with a history of radiation therapy to a "dog-leg" field including the lumbar vertebrae underwent MR imaging of their lumbar spine using a 0.5 Tesla magnet. Five healthy subjects and two nonirradiated patients were imaged as well. The intervals from the beginning of radiotherapy to MRI examination varied from 1.5 to 52 months, and the radiation dose ranged from 3000-4200 cGy. The T2 relaxation times of the lumbar vertebral bone marrow and subcutaneous fat were calculated for each subject. Postirradiation bone marrow in irradiated seminoma patients exhibited significantly longer T2 relaxation times than nonirradiated bone marrow in controls (71.1 vs. 63.6 ms, p = 0.047, t-test). The differences between the T2 relaxation times of bone marrow and subcutaneous fat for each subject allowed for even better differentiation between irradiated patients and controls (10.4 vs. 0.4 ms, p = 0.0004, t-test). Postirradiation bone marrow had significantly longer T2 relaxation times than subcutaneous fat in irradiated patients (N = 10, 71.1 vs. 60.7 ms, p = 0.00009, t-test), while nonirradiated bone marrow had T2 relaxation times not statistically different from subcutaneous fat in nonirradiated subjects (N = 7, 63.6 vs. 63.2 ms). Measurements of T2 relaxation times of bone marrow enabled us to differentiate between irradiated seminoma patients and controls. Postirradiation bone marrow undergoes late radiation effects resulting in longer T2 relaxation times than nonirradiated bone marrow and subcutaneous fat. PMID- 9201680 TI - Monitoring brain tumor response to therapy using MRI segmentation. AB - The performance evaluation of a semi-supervised fuzzy c-means (SFCM) clustering method for monitoring brain tumor volume changes during the course of routine clinical radiation-therapeutic and chemo-therapeutic regimens is presented. The tumor volume determined using the SFCM method was compared with the volume estimates obtained using three other methods: (a) a k nearest neighbor (kNN) classifier, b) a grey level thresholding and seed growing (ISG-SG) method and c) a manual pixel labeling (GT) method for ground truth estimation. The SFCM and kNN methods are applied to the multispectral, contrast enhanced T1, proton density, and T2 weighted, magnetic resonance images (MRI) whereas the ISG-SG and GT methods are applied only to the contrast enhanced T1 weighted image. Estimations of tumor volume were made on eight patient cases with follow-up MRI scans performed over a 32 week interval during treatment. The tumor cases studied include one meningioma, two brain metastases and five gliomas. Comparisons with manually labeled ground truth estimations showed that there is a limited agreement between the segmentation methods for absolute tumor volume measurements when using images of patients after treatment. The average intraobserver reproducibility for the SFCM, kNN and ISG-SG methods was found to be 5.8%, 6.6% and 8.9%, respectively. The average of the interobserver reproducibility of these methods was found to be 5.5%, 6.5% and 11.4%, respectively. For the measurement of relative change of tumor volume as required for the response assessment, the multi-spectral methods kNN and SFCM are therefore preferred over the seedgrowing method. PMID- 9201682 TI - Effects of vasodilators on the signal intensity of perfluorocarbon monitored by in vivo 19F-NMR spectroscopy. AB - The effects of vasodilators on peripheral vessels were examined by monitoring the 19F-NMR signal of perfluorocarbon in vivo. Nitroglycerin, a venodilator that acts mainly on venous smooth muscle, increased the signal intensity of FC-43, whereas hydralazine, a typical arteriolar dilator that acts on arteriolar smooth muscle, decreased the signal intensity. These results indicate that the in vivo effects of vasodilators on smooth muscles of the venous and arterial systems are reflected by their effects on the signal intensity of FC-43. PMID- 9201683 TI - Changes in 31P-relaxation times during organ preservation: observations on cold stored human donor livers. AB - During cold preservation for transplantation the tissue hydration state changes. It is not known whether such changes lead to altered relaxation times of 31P nuclei with potential consequences for the quantification of tissue metabolites. Therefore, 31P spectroscopic and proton T1 relaxometric measurements were performed on 42 isolated human donor livers shortly before implantation. The results demonstrate that 31P T1 relaxation times change during preservation for clinical transplantation, thus quantification of tissue metabolites in cold stored donor livers may be in part dependent on the tissue hydration state. Furthermore, it appeared that changes in tissue hydration state especially affect the physico-chemical characteristics of the intracellular fluid compartment. This study indicates that reliable spectroscopic quantification of tissue metabolites, particularly during sequential spectroscopic measurements in cold stored donor organs is best warranted under fully relaxed conditions. PMID- 9201685 TI - An application of NMR microimaging to investigate nitrogen fixing root nodules. AB - Various techniques to obtain high-resolution NMR images (voxel size down to 39 x 39 x 250 microns) of nitrogen-fixing root nodules from soybean [Glycine max (Merr.)] and peanut (Arachis hypogaea L.) are compared. We describe the artefacts arising from changes in the magnetic susceptibility throughout the sample and how these can be minimised. A series of T1 (TR = 220 to 3020 ms) and T2 (TE = 9.3 to 33.6 ms) weighted images are presented. From these it has been possible to locate the oxygen diffusion barrier. A possible interpretation in terms of nodule biochemistry and physiology are given. The data and parameters presented are shown to serve as a basis for more extensive investigations of root nodules (e.g., the oxygen diffusion barrier or the mechanisms driving the regulation of the oxygen concentration in the infected zone by leghemoglobin) by NMR microimaging. PMID- 9201684 TI - ESR imaging of the rat brain with a nitroxide radical perfused by in vivo microdialysis. AB - We report here our investigation of the spatial distribution of free radicals using an electron spin resonance (ESR)-imaging system combined with an in vivo brain microdialysis method, which was performed in the resonator of the ESR imaging system. A nonmagnetic cannula, newly developed in this study, was used for the perfusion of the exogenous free radicals agent. A nitroxide, 3-carbamoyl 2,2,5,5-tetramethylpyrrolidine-1-oxyl (carbamoyl PROYXL), was used as the imaging agent in saline solution at a concentration of 0.3 M, which was perfused into the right caudate putamen of the rat at 2 microliters/min by a microinfusion pump. Two-dimensional ESR projection of the Z-X plane, which was clearly distinguished (about phi 10 mm) from the nonperfused brain area, was obtained 6 h after the beginning of perfusion of carbamoyl PROXYL. The present method is considered to be a useful tool to introduce stable free radicals into a specific area of the brain. PMID- 9201686 TI - Design of a biplanar gradient coil using a genetic algorithm. AB - A biplanar z-gradient coil has been designed using a genetic algorithm, and its efficiency for producing a gradient along the axis of a solenoid magnet compared to that of a conventional Maxwell coil set. Coils of 21.8 cm by 20.9 cm area and 10 cm separation give 0.37 m Tm-1 A-1 with standard and maximum deviations of 2.6 and 13.1% of this value over an optimised cuboid region of 12 by 15 by 1.8 cm. The experimentally useable linear volume extends beyond this to 50% of the separation between the planes. Design data are also given for a transverse gradient set. PMID- 9201687 TI - Localized pleural mesothelioma: CT and MR findings. AB - A case of localized pleural mesothelioma inducing hypoglycemic coma is presented. CT and MR findings are described. T1-weighted MR images demonstrated the mass of slightly high signal intensity with lesions of signal void. Varying degrees of T2 shortening were shown on T2-weighted MR images. Coronal MR imaging was useful for assessing the relationship between the diaphragm and lesions in the lower chest. PMID- 9201688 TI - Reproducibility of metabolite peak areas in 1H MRS of the human brain. PMID- 9201689 TI - Quantitative assessment of tuftsin receptor expression and second messenger during in vitro differentiation of peripheral blood derived monocytes of leprosy patients. AB - Tuftsin, a tetrapeptide (Thr-Lys-Pro-Arg) is known to potentiate the immunogenic activity of antigen-fed macrophages. The present study describes the mechanism of action of tuftsin in leprosy patients throughout the spectrum of the disease in vitro as a function of culture age in terms of (A) involvement of second messengers cAMP, cGMP and [Ca2+]i and (B) number of tuftsin binding sites/and their relative affinities on the monocytes/macrophages. There is apparently no direct involvement of either cAMP or cGMP while comparing the stimulated and unstimulated cultures during in vitro differentiation of monocytes (days 1, 3 and 7) or with the spectrum of the disease. Inhibition of superoxide anion release either by verapamil or with Quin 2 clearly demonstrated the involvement of [Ca2+]i as a second messenger during activation of monocytes/macrophages with tuftsin. Scatchard analysis of radiolabelled tuftsin binding data showed only one type of tuftsin receptor (low affinity) on BL/ LL monocytes/macrophages and normal and BT/TT cultures showed a gradual change in receptor number and affinities (low to high) with the maturation of monocytes to macrophages in contrast to BL/LL groups which displayed significantly less number of receptors. This study elicits a model which depicts that the biological responses/metabolic functions of early monocytes of normal and BT/TT gradually increase with the age of the culture till day 3 and tapers off thereafter in the older (day 7) cultures, whereas the monocytes/macrophages of BL/LL group are metabolically active only on day 1. The present study thereby implies that the clearance of leprosy bacilli from lepromatous leprosy lesions as a consequence of local or systemic immunotherapy (in the present study, the macrophage modulation by tuftsin) depends on the influx of new competent macrophages, rather than the local activation of resident lepromatous macrophages. PMID- 9201690 TI - Transient Ca2+ changes in endothelial cells induced by low doses of reactive oxygen species: role of hydrogen peroxide. AB - Cultured human and rat endothelial cells were used to study cellular toxicity and Ca2+ signalling upon exposure to reactive oxygen species. Superoxide and hydrogen peroxide (O2.-/H2O2) were produced by the hypoxanthine/xanthine oxidase system (HX/XO) and caused intracellular Ca2+ concentration ([Ca2+]i) to rise steadily when activities above 2 mU/ml were used. These Ca2+ increases were also measured when the glucose/glucose oxidase (G/GO) system above 5 mU/ml was used to produce hydrogen peroxide (H2O2). Gross morphological changes appeared to parallel elevated [Ca2+]i levels preceding cell death. However, when HX/XO or G/GO were used at non toxic doses rapid and transient changes in [Ca2+]i were measured. These treatments did not alter subsequent receptor mediated Ca2+ signalling induced by ATP (10 microM) or histamine (100 microM). Superoxide dismutase (50 U/ml), which dismutates O2.- into H2O2 also had no influence, whereas catalase (50 U/ml), which removes H2O2, completely diminished transient [Ca2+]i responses. H2O2 added directly was able to induce similar Ca2+ transients when concentrations of at least 500 microM were used. Buffering trace amounts of iron (o-phenanthroline; 200 microM) in order to inhibit .OH radical formation was not effective to alter Ca2+ changes. Experiments performed in Ca(2+)-free buffer showed a similar rise in [Ca2+]i and readdition of Ca2+ to the extracellular medium indicated the activation of store operated Ca2+ entry. Blocking Ca(2+) ATPases of the endoplasmatic reticulum with thapsigargin (1 microM) inhibited ROS induced transient increases and cells preincubated with pertussis toxin (200 nM) showed unchanged Ca2+ transients after exposure to both enzyme systems. Phospholipase C inhibitor U73122 (2 microM) effectively reduced hydrogen peroxide induced emptying of intracellular stores. Taken together, we demonstrate that enzymatically produced non-toxic H2O2 rather than O2.- or .OH causes calcium signalling from thapsigargin sensitive stores, and activates store operated Ca2+ entry at least partially by activating phospholipase C. These changes clearly differ from pathological 'oxidative stress' associated with a progressive increase in [Ca2+]i. PMID- 9201691 TI - Formation of advanced glycation end (AGE) products in diabetes: prevention by pyruvate and alpha-keto glutarate. AB - Glycation of proteins and their subsequent structural and functional modifications have been ascribed to play a prominent role in the pathogenesis of several secondary complications of diabetes, such as cataract and retinopathy. In addition, it plays a role in the generalized ageing process as well. Investigations have been conducted to explore the possibility of preventing the above process by use of pyruvate and alpha-keto glutarate as representatives of physiologically compatible keto acids. The results demonstrate that both these compounds are effective in preventing the initial glycation reaction as well as the formation of AGE products. Both these compounds also inhibit the generation of high molecular weight aggregates associated with cataract formation. Mechanistically, the preventive effects appear to be due to (1) competitive inhibition of glycation by the keto acids and (2) the antioxidant (radical scavenging) properties of these compounds. The results are hence considered useful from the point of view of developing these and other keto acid derivatives as pharmacological agents useful in preventing glycation related protein changes and consequent tissue pathological manifestations. PMID- 9201692 TI - Inhibitors of in vitro mineralization from flexor tendons of rabbits and their role in biological mineralization. AB - Studies demonstrate that flexor tendons contain loosely associated biomolecules which inhibit its mineralization under physiological conditions. Based upon their molecular weights, these inhibitory biomolecules, could be classified into two categories, having molecular weights less than and greater than 13,000 daltons. The main inhibitory biomolecule was found to be an acidic polypeptide having molecular weight of 12,400 daltons. PMID- 9201694 TI - Nuclease susceptibility of the rat liver satellite DNA-containing chromatin decreases with age. AB - Nuclease susceptibility of the satellite DNA-containing chromatin of the liver of young (18 +/- 2 weeks) and old (100 +/- 5 weeks) rats was analysed using nick translated rat 185 bp satellite I DNA fragment cloned in pBR322. With increasing concentration of DNaseI and micrococcal nuclease (MNase), multimeric forms of the satellite ladder gradually disappear in both the ages. The rate of disappearance is faster in young rats as compared to old ones. Such age-dependent decrease in the susceptibility of satellite DNA-containing chromatin reflects its condensation towards heterochromatization in old age. PMID- 9201693 TI - Calreticulin inhibits glucocorticoid- but not cAMP-sensitive expression of tyrosine aminotransferase gene in cultured McA-RH7777 hepatocytes. AB - Calreticulin is a ubiquitously expressed Ca2+ binding protein of the endoplasmic reticulum which inhibits DNA binding and transcriptional activation by steroid hormone receptors. In this study the effects of calreticulin on tyrosine aminotransferase (TAT) gene expression in cultured McA-RH7777 hepatocytes was investigated. McA-RH7777 cells were stably transfected with calreticulin expression vector to generate cells overexpressing the protein. The transcriptional activity of the TAT gene, which is glucocorticoid-sensitive and cAMP-dependent, was investigated in the mock transfected McA-RH7777 and in cells overexpressing calreticulin (designated McA-11 and McA-17). In the presence of dexamethasone or the cAMP analog (CTP-cAMP) expression of the TAT gene was induced in mock transfected McA-RH7777 cells by approximately 4.5 and 5 fold, respectively. In McA-11 and McA-17 cells, overexpressing calreticulin, glucocorticoid-sensitive expression of the TAT gene was significantly inhibited, however, the CTP-cAMP-dependent expression of the TAT gene was not affected. The ability of calreticulin to inhibit glucocorticoid-sensitive TAT gene expression but not the cAMP-dependent expression of the gene suggests that the protein affects specifically the action of transcription pathways involving steroid receptors or transcription factors containing KxFF(K/R)R-like motifs. Calreticulin may play an important role in the regulation of glucocorticoid sensitive pathway of expression of the hepatocytes specific genes during development. PMID- 9201696 TI - The effects of anaplerotic substrates on D-3-hydroxybutyrate metabolism in the heart. AB - In this study the effects of propionate, L-valine, L-isoleucine, and DL methionine on the metabolism of D-3-hydroxybutyrate (D-3-HB) were investigated in the isolated perfused non-working rat heart. Propionate inhibited the utilization (the total removal of D-3-HB by the heart) but stimulated the oxidation of D-3 HB. The degree of D-3-HB inhibition was dependent on the concentrations of propionate and D-3-HB. Furthermore, increasing the concentration of DL hydroxybutyrate (DL-3-HB) to 16 or 30 mM abolished the inhibitory effect of propionate (4 mM). Whereas increasing the perfusion pressure from 40-80 mmHg stimulated the utilization and the oxidation of D-3-HB; propionate (4 mM) severely inhibited the utilization of D-3-HB at 40 and 80 mmHg, when DL-3-HB was 5 mM. On the other hand insulin (2 mU .ml-1) stimulated the utilization and the oxidation of D-3-HB at perfusion pressure of 40 mmHg, but showed no effect at 80 mmHg. Insulin was unable to overcome the inhibitory effect of propionate. Propionate improved the oxidation but inhibited the utilization of D-3-HB, while L-valine and L-isoleucine showed no effects on the utilization and the oxidation of D-3-HB. DL-methionine increased the utilization of D-3-HB by 14% without noticeable effects on the oxidation of D-3-HB. None of these anaplerotic substrates were suitable to ameliorate the utilization of D-3-HB. PMID- 9201697 TI - Heme metabolism in promastigotes of Leishmania donovani. AB - Promastigotes of Leishmania donovani (Dd-8 strain) showed presence of important key enzymes of heme synthesizing (delta-aminolevulinic acid synthase and ferrochelatase) and degrading (heme oxygenase and biliverdin reductase) systems, classical leishmanicidal drugs viz allopurinol, amphotericin B, pentamidine and CDRI compound 93/202 inhibited the heme oxygenase activity of the parasite, whereas, delta-aminolevulinic acid synthase activity practically remained unaffected. The Km, Vmax and pH values of heme oxygenase of promastigotes were found to be 1666 microM hemin, 625 nmol of bilirubin formed h-1 mg protein-1 and 7.5 respectively. The findings suggest the presence and importance of heme metabolism in the de novo synthesis of different hemoproteins of the Leishmania parasite as well as the detoxification and its defence against biological insults. PMID- 9201695 TI - Inhibition of human lymphocyte function by organic solvents. AB - We studied the direct effect of reactive hydroxyl precursors and inhibitors on CD4+ T-cell function. We used hydrogen peroxide plus ferrous chloride as the hydroxyl radical-generating system and di-methyl sulphourea, di-methyl sulfoxide, pyrrolidine dithiocarbonate, methanol, and ethanol, at a noncytotoxic concentration, as inhibitors. The immune parameter studies were proliferation and interleukin-2 production by peripheral blood lymphocytes stimulated with anti-CD3 antibody, phytohemagglutinin and alloantigens; proliferation, interleukin-2 production and mRNA expression of interleukin-4 and interferon gamma by allogeneic CD4+ T-cell clones stimulated with alloantigens. The results show that lymphocytes produce significant amounts of reactive oxygen species as measured by malondialdehyde produced in cultures. The hydroxyl radical-generating system did not change any of the cellular responses studied although it doubled Malondialdehyde production. Hydroxyl radical scavengers significantly inhibited all responses at doses that didn't significantly decrease malondialdehyde production. DNA analysis failed to show evidence for apoptosis. CONCLUSION: Hydroxyl radical scavengers inhibit lymphocyte mitogenesis by a process that is independent of scavenging hydroxyl radicals. PMID- 9201698 TI - Effect of cations on the tyrosine kinase activity of the insulin receptor: inhibition by fluoride is magnesium dependent. AB - We have recently reported that fluoride interacts directly with the insulin receptor, which causes inhibition of its phosphotransferase activity. The inhibitory effect of fluoride on phosphotransferase activity is not due to the formation of complexes with aluminium and occurs in the absence of alterations to the binding of ATP or insulin. In this report we substantiate that the tyrosine kinase activity of insulin receptors partially purified from rat skeletal muscle shows a strict requirement of Mg2+ ions (Ka near 11 mM). This effect of Mg2+ was inhibited in a competitive manner by Mn2+, which is compatible with competition of both divalent ions for binding sites. The inhibition of tyrosine kinase activity caused by fluoride was dependent on the concentration of Mg2+ in the medium and no inhibitory effect was detected at low concentrations of Mg2+. Moreover, the addition of increasing concentrations of Mn2+ in the presence of a constant high concentration of Mg2+, led to a gradual decrease in the inhibitory effect of fluoride. These results indicate that the Mg-insulin receptor complex is the major fluoride-susceptible form. Based on the characteristics of the inhibition of tyrosine kinase shown by fluoride it might be proposed that its action is exerted by the formation of multi-ionic MgF complexes analogous to Pi, which bind to the insulin receptor kinase. PMID- 9201700 TI - Magnesium: effects on reperfusion arrhythmias and membrane potential in isolated rat hearts. AB - The effects of Mg2+ concentration (Mg2+o, 0, 1.2, 2.4, and 4.8 mM) on the incidence of reperfusion arrhythmias and on the cellular electrical activity were studied in spontaneously beating rat hearts. The surface electrogram and the membrane potential were recorded in control conditions, during 10 min of regional ischemia (ligature of the left anterior descending coronary artery), and on reflow. Changes in Mg2+o did not alter action potential morphology but the depolarization induced by ischemia decreased with increasing Mg2+o. In hearts perfused with Mg2+ free solution or 1.2 mM subthreshold delayed afterdepolarizations (DADs) were often detected during ischemia. Moreover, DADs could be identified as initial events in the production of extrabeats or tachycardia appearing on reperfusion under these conditions. Chaotic electrical activity during fibrillation precluded the observation of DADs. The overall incidence (100%) and severity of ventricular tachyarrhythmias (80% tachycardia and fibrillation) was similar in both groups. At high Mg2+o, subthreshold DADs were occasionally observed during ischemia and often on reperfusion where they did not lead to the development of overt arrhythmias. Consequently, the incidence, severity, and duration of arrhythmic episodes on reflow was markedly reduced. Raising Mg2+ only on reperfusion did not prevent the development of arrhythmias, whose morphology in the intracellular recordings was similar to that found in hearts perfused without Mg2+ or with 1.2 mM. The recovery of sinus rhythm after 10 min of reperfusion was linearly related to Mg2+o. Our data strengthen the view that reperfusion arrhythmias belong to the Ca2+ mediated non reentrant type and suggest that Mg2+ counteracts these arrhythmias by depressing cytosolic Ca2+ oscillations. Besides, it appears that raising Mg2+o reduces ischemic K+o accumulation. The resulting changes in resting potential could contribute to lower DADs amplitude and thus decrease the arrhythmogenic potential of the Ca2+i oscillations induced by reperfusion. PMID- 9201701 TI - Similar nature of inhibition of mitochondrial respiration of heart tissue and malignant cells by methylglyoxal. A vital clue to understand the biochemical basis of malignancy. AB - The effect of methylglyoxal on the oxygen consumption of mitochondria of heart and of several other organs of normal animals of different species has been tested. The results indicate that methylglyoxal (3.5 mM) strongly inhibits ADP stimulated alpha-oxoglutarate and malate plus pyruvate-dependent respiration of exclusively heart mitochondria of normal animals of different species. Whereas, with the same substrates, but at a higher concentration of methylglyoxal (7.5 mM), the respiration of mitochondria of other organs of normal animals is not inhibited. Methylglyoxal also inhibits the respiration of slices of rat and toad hearts. But this inhibition is less pronounced. However, methylglyoxal (15 mM) fails to have any effect on perfused toad heart. Using rat heart mitochondria as a model, the effect of methylglyoxal on the oxygen consumption was also tested with different respiratory substrates, electron donors at different segments of the mitochondrial respiratory chain and site-specific inhibitors to identify the specific respiratory complex which might be involved in the inhibitory effect of methylglyoxal. The results strongly suggest that methylglyoxal inhibits the electron flow through complex I of rat heart mitochondrial respiratory chain. Moreover, lactaldehyde (0.6 mM), a catabolite of methylglyoxal, can exert a protective effect on the inhibition of rat heart mitochondrial respiration by methylglyoxal (2.5 mM). The effect of methylglyoxal on heart mitochondria as described in the present paper is strikingly similar to the results of our previous work with mitochondria of Ehrlich ascites carcinoma cells and leukemic leukocytes. We have recently proposed a new hypothesis on cancer which suggests that excessive ATP formation in cells may lead to malignancy. The above mentioned similarity apparently provides a solid experimental foundation for the proposed hypothesis which has been discussed. PMID- 9201699 TI - Analysis of the in vitro effect of exogenous nitric oxide on human lymphocytes. AB - We investigated the role of endogenous or exogenous nitric oxide (NO) on human lymphocyte function. We used sodium nitroprusside, nitroglycerine, S-nitroso-N acetylpenicillamine, sodium nitrite and S-nitroso-L-glutathione as NO-generating compounds. All agents were used at doses that do not produce direct cytotoxicity as measured by trypan blue exclusion as well as chromium-51 release assay. The immune responses examined were peripheral blood lymphocytes (PBL) proliferation and IL-2 production after activation with OKT3 and PHA; allogeneic mediated proliferation and cell mediated cytotoxicity (CML) in MLR; IgG and IgM production after PBL activation with Con-A; proliferation and expression of IFN-gamma and IL 4 mRNA after activation of allogeneic CD4+T cell clones. Cytokine mRNA expression was measured by reverse transcriptase PCR. Our results show that proliferating lymphocytes do not produce a detectable amount of NO as measured by the Griess reaction. In separate experiments, the addition of NG-monomethyl-L-arginine (L NMMA) did not affect lymphocyte proliferation. Sodium nitroprusside and nitroglycerine exerted a dose dependent antimitogenic effect, inhibited cytokine production and expression, CML generation and antibody production. DNA gel electrophoresis showed no evidence for enhanced programmed cell death. The antimitogenic effect could not be blocked by the NO scavengers, hemoglobin or methylene blue. In contrast, the other nitric oxide generating compounds did not inhibit lymphocyte mitogenesis. The results suggest that human lymphocytes do not produce appreciable amounts of NO to affect lymphocyte mitogenesis. Sodium nitroprusside and nitroglycerine have a potent but nonspecific immunoinhibitory effect on human lymphocyte function by a mechanism other than NO production. In addition, pharmacological levels of NO do not inhibit human lymphocyte mitogenesis. PMID- 9201702 TI - Chromatographic separation of DNA dependent RNA polymerases and molecular properties of RNA polymerase II from a Leishmania Spp. AB - Multiple forms of DNA-dependent RNA polymerases have been isolated and characterized from Leishmania strain UR6 promastigotes. RNA polymerases from this organism fail to resolve into multiple forms by conventional chromatography on DEAE-Sephadex A25, but could be separated by a modification of the method using CM-Sephadex C25. The CM-Sephadex bound enzyme is resistant to alpha-amanitin even up to a concentration of 250 micrograms/ml. The activity which flows through CM Sephadex further resolves into two forms upon chromatography on DEAE-Sephadex A25. These forms are sensitive to alpha-amanitin to different extent. Enzyme activity in peak I is 50% inhibited by 3 micrograms/ml and in peak II by 50 micrograms/ml of the drug respectively. The enzyme in peak I has been further purified by heparin agarose and fast performance liquid chromatography (FPLC) on MonoQ. The enzyme has Stoke's radius of 70 A, a sedimentation coefficient of 17.6S and an f/fo of 1.35. Analysis of ammonium sulfate and metal ion optima of the enzyme in peak I, relative activities with Mn+2 versus Mg+2 and template specificities gave results similar to those reported for other type II RNA polymerases in eukaryotes. The MonoQ purified enzyme resolves into 16 polypeptides on denaturing polyacrylamide gel and densitometric analysis suggests that 9 major bands are present in the stoichiometry expected of RNA polymerase subunits having molecular weights: 154000; 104000; 77000; 64000; 52000; 48000; 46000; 45000 and 39000 respectively. PMID- 9201703 TI - Phosphorylation of protein phosphatase-1 isoforms by cdc2-cyclin B in vitro. AB - Protein Phosphatase-1 is phosphorylated in vitro by cdc2-cyclin B (Villa-Moruzzi, FEBS Lett 304: 211-215, 1992). In the present study we show that all the three Phosphatase-1 isoforms, alpha, gamma 1, delta, are phosphorylated by cdc2-cyclin B. Phosphorylation is specific for this kinase and involves a C-terminal Thr. This site is most likely Thr 320 in alpha (shown by others to be phosphorylated also by cdc2-cyclin A). Such Thr is conserved in gamma 1, delta and in the testis specific gamma 2, and is the only Thr that fits the cdc2-consensus sequence in the C-terminal region. Phosphorylation of Phosphatase-1 purified from skeletal muscle, which is a mixture of the alpha, gamma 1 and delta isoforms, is up to 0.4 mol/mol and induces 30-35% enzyme inactivation. Following tryptic proteolysis each isoform yields a distinct phosphopeptide map. This is in agreement with the different sequences of the isoforms in the C-terminal regions and may be useful to distinguish the isoforms in extracts from metabolically-labelled cells. Our results suggest that all the Phosphatase-1 isoforms may be potentially regulated at M-phase. PMID- 9201704 TI - The Wilms tumor protein is persistently associated with the nuclear matrix throughout the cell cycle. AB - In order to investigate the subnuclear interactions of the WT1 gene product, nuclear fractionation analyses were performed with human osteosarcoma HOS and myelogenous leukemia K562 cells. The WT1 protein was tightly associated with the nucleus and was resistant to high-salt or detergent extraction and DNase I digestion. Both the expression level and stability of WT1 and its resistance to high salt and DNase I treatments remained constant during the cell cycle. In addition, human WT1 ectopically expressed in mouse NIH3T3 cells was also resistant to these treatments. These results suggest that WT1 functions in tight association with the nuclear matrix. PMID- 9201705 TI - Increase in calcium content and Ca(2+)-ATPase activity in the brain of fasted rats: comparison with different ages. AB - The effect of fasting on calcium content and Ca(2+)-ATPase activity in the brain tissues of 5 weeks and 50 weeks old rats was investigated. Brain calcium content and Ca(2+)-ATPase activity in the microsomal and mitochondrial functions of the brain homogenate from young and elderly rats were significantly increased by overnight-fasting. These increases were appreciably restored by a single oral administration of glucose solution (400 mg/100 g body weight) to fasted rats. In comparison with young and elderly rats, brain calcium content and microsomal Ca(2+)-ATPase activity were significantly elevated by increasing ages. The effect of ageing was not seen in the brain mitochondrial Ca(2+)-ATPase activity. When calcium (50 mg/100 g) was orally administered to young and elderly rats, brain calcium content was significantly elevated. The calcium administration-induced increase in brain calcium content was greater in elderly rats than in young rats. Also, calcium administration caused a significant increase in Ca(2+)-ATPase activity in the microsomal and mitochondrial fractions of brain homogenates from young rats. In aged rats, the microsomal Ca(2+)-ATPase activity was not further enhanced by calcium administration, although the mitochondrial enzyme activity was significantly raised. The present study demonstrates that the fasting-induced increase in brain calcium content is involved in Ca(2+)-ATPase activity raised in the brain microsomes and mitochondria of rats with different ages, supporting a energy-dependent mechanism in brain calcium accumulation. PMID- 9201706 TI - Androgen deprivation causes up-regulation of androgen receptor transcript in the rat prostate. AB - We have studied the effect of androgenic deprivation on the level of androgen receptor transcript in the rat ventral prostate. The rats were treated with estradiol benzoate, flutamide and [D Trp6, des Gly10]gonadotropin releasing hormone (GnRH) for different time periods. These treatments produced a significant decrease in the weight of prostate. Total RNA isolated from the ventral prostates was hybridized with the cDNA probe for androgen receptor. Densitometric analysis of the autoradiographic signal revealed a rise in the level of androgen receptor RNA following treatment of rats with estradiol benzoate and flutamide. Treatment of rats with [D Trp6, des Gly10] GnRH brought about a transient rise in the level of androgen receptor RNA. Thus, our results indicate that androgenic deprivation up-regulates the level of androgen receptor transcript. PMID- 9201707 TI - Functional domains in nuclear import factor p97 for binding the nuclear localization sequence receptor and the nuclear pore. AB - The interaction of the nuclear protein import factor p97 with the nuclear localization sequence (NLS) receptor, the nuclear pore complex, and Ran/TC4 is important for coordinating the events of protein import to the nucleus. We have mapped the binding domains on p97 for the NLS receptor and the nuclear pore. The NLS receptor-binding domain of p97 maps to the C-terminal 60% of the protein between residues 356 and 876. The pore complex-binding domain of p97 maps to residues 152-352. The pore complex-binding domain overlaps the Ran-GTP- and Ran GDP-binding domains on p97, but only Ran-GTP competes for docking in permeabilized cells. The N-ethylmaleimide sensitivity of the p97 for docking was investigated and found to be due to inhibition of p97 binding to the pore complex and to the NLS receptor. Site-directed mutagenesis of conserved cysteine residues in the pore- and receptor-binding domains identified two cysteines, C223 and C228, that were required for p97 to bind the nuclear pore. Inhibition studies on docking and accumulation of a NLS protein provided additional evidence that the domains identified biochemically are the functional domains involved in protein import. Together, these results suggest that Ran-GTP dissociates the receptor complex and prevents p97 binding to the pore by inducing a conformational change in the structure of p97 rather than simple competition for binding sites. PMID- 9201708 TI - Centromere position in budding yeast: evidence for anaphase A. AB - Although general features of chromosome movement during the cell cycle are conserved among all eukaryotic cells, particular aspects vary between organisms. Understanding the basis for these variations should provide significant insight into the mechanism of chromosome movement. In this context, establishing the types of chromosome movement in the budding yeast Saccharomyces cerevisiae is important since the complexes that mediate chromosome movement (microtubule organizing centers, spindles, and kinetochores) appear much simpler in this organism than in many other eukaryotic cells. We have used fluorescence in situ hybridization to begin an analysis of chromosome movement in budding yeast. Our results demonstrate that the position of yeast centromeres changes as a function of the cell cycle in a manner similar to other eukaryotes. Centromeres are skewed to the side of the nucleus containing the spindle pole in G1; away from the poles in mid-M and clustered near the poles in anaphase and telophase. The change in position of the centromeres relative to the spindle poles supports the existence of anaphase A in budding yeast. In addition, an anaphase A-like activity independent of anaphase B was demonstrated by following the change in centromere position in telophase-arrested cells upon depolymerization and subsequent repolymerization of microtubules. The roles of anaphase A activity and G1 centromere positioning in the segregation of budding yeast chromosomes are discussed. The fluorescence in situ hybridization methodology and experimental strategies described in this study provide powerful new tools to analyze mutants defective in specific kinesin-like molecules, spindle components, and centromere factors, thereby elucidating the mechanism of chromosome movement. PMID- 9201709 TI - Nanomolar concentrations of nocodazole alter microtubule dynamic instability in vivo and in vitro. AB - Previous studies demonstrated that nanomolar concentrations of nocodazole can block cells in mitosis without net microtubule disassembly and resulted in the hypothesis that this block was due to a nocodazole-induced stabilization of microtubules. We tested this hypothesis by examining the effects of nanomolar concentrations of nocodazole on microtubule dynamic instability in interphase cells and in vitro with purified brain tubulin. Newt lung epithelial cell microtubules were visualized by video-enhanced differential interference contrast microscopy and cells were perfused with solutions of nocodazole ranging in concentration from 4 to 400 nM. Microtubules showed a loss of the two-state behavior typical of dynamic instability as evidenced by the addition of a third state where they exhibited little net change in length (a paused state). Nocodazole perfusion also resulted in slower elongation and shortening velocities, increased catastrophe, and an overall decrease in microtubule turnover. Experiments performed on BSC-1 cells that were microinjected with rhodamine-labeled tubulin, incubated in nocodazole for 1 h, and visualized by using low-light-level fluorescence microscopy showed similar results except that nocodazole-treated BSC-1 cells showed a decrease in catastrophe. To gain insight into possible mechanisms responsible for changes in dynamic instability, we examined the effects of 4 nM to 12 microM nocodazole on the assembly of purified tubulin from axoneme seeds. At both microtubule plus and minus ends, perfusion with nocodazole resulted in a dose-dependent decrease in elongation and shortening velocities, increase in pause duration and catastrophe frequency, and decrease in rescue frequency. These effects, which result in an overall decrease in microtubule turnover after nocodazole treatment, suggest that the mitotic block observed is due to a reduction in microtubule dynamic turnover. In addition, the in vitro results are similar to the effects of increasing concentrations of GDP-tubulin (TuD) subunits on microtubule assembly. Given that nocodazole increases tubulin GTPase activity, we propose that nocodazole acts by generating TuD subunits that then alter dynamic instability. PMID- 9201710 TI - Functional analysis of the interaction between Afr1p and the Cdc12p septin, two proteins involved in pheromone-induced morphogenesis. AB - Saccharomyces cerevisiae mating pheromones induce production of Afr1p, a protein that negatively regulates pheromone receptor signaling and is required for normal formation of the projection of cell growth that becomes the site of cell fusion during conjugation. Afr1p interacts with Cdc12p, which belongs to a family of filament-forming proteins termed septins that have been studied primarily for their role in bud morphogenesis and cytokinesis. The significance of the interaction between Afr1p and Cdc12p was tested in this study by examining the effects of AFR1 mutations that destroy the Cdc12p-binding domain. The results demonstrate that sequences in the C-terminal half of Afr1p are required for interaction with Cdc12p and for proper localization of Afr1p to the base of the mating projection. However, the Cdc12p-binding domain was not required for regulation of receptor signaling or for mating projection formation. This result was surprising because cells carrying a temperature-sensitive cdc12-6 mutation were defective in projection formation, indicating a role for Cdc12p in this process. Although the Cdc12p-binding domain was no essential for Afr1p function, this domain did improve the ability of Afr1p to promote morphogenesis, suggesting that the proper localization of Afr1p is important for its function. PMID- 9201711 TI - Perlecan regulates Oct-1 gene expression in vascular smooth muscle cells. AB - Vascular smooth muscle cells (SMCs) are very quiescent in the mature vessel and exhibit a remarkable phenotype-dependent diversity in gene expression that may reflect the growth responsiveness of these cells under a variety of normal and pathological conditions. In this report, we describe the expression pattern of Oct-1, a member of a family of transcription factors involved in cell growth processes, in cultured and in in vivo SMCs. Oct-1 mRNA was undetectable in the contractile-state in vivo SMCs; was induced upon disruption of in vivo SMC extracellular matrix interactions; and was constitutively expressed by cultured SMCs. Oct-1 transcripts were repressed when cultured SMCs were plated on Engelbreth-Holm-Swarm tumor-derived basement membranes (EHS-BM) but were rapidly induced after disruption of SMC-EHS-BM contacts; reexpression was regulated at the transcriptional level. To identify the EHS-BM component involved in the active repression of Oct-1 mRNA expression, SMCs were plated on laminin, type IV collagen, fibronectin, or perlecan matrices. Oct-1 mRNA levels were readily detectable when SMCs were cultured on matrices composed of laminin, type IV collagen, or fibronectin but were repressed when SMCs were cultured on perlecan matrices. Finally, the Oct-1-suppressing activity of EHS-BM was sensitive to heparinase digestion but not to chondroitinase ABC or hyaluronidase digestion, suggesting that the heparan sulfate side chains of perlecan play a biologically important role in negatively regulating the expression of Oct-1 transcripts. PMID- 9201712 TI - The role of inhibitory phosphorylation of CDC2 following DNA replication block and radiation-induced damage in human cells. AB - It has been suggested that the survival response of p53 defective tumor cells to agents that inhibit DNA replication or damage DNA may be largely dependent on cell cycle checkpoints that regulate the onset of mitosis. In human cells, the mitosis-inducing kinase CDC2/cyclin B is inhibited by phosphorylation of threonine-14 and tyrosine-15, but the roles of these phosphorylations in enforcing checkpoints is not known. We have investigated the situation in a human cervical carcinoma cell line (HeLa cells) and found that low level expression of a mutant nonphosphorylatable form of CDC2 abrogates regulation of the endogenous CDC2/cyclin B. Disruption of this pathway is toxic and renders cells highly sensitive to killing by DNA damage or by inhibition of DNA replication. These findings establish the importance of inhibitory phosphorylation of CDC2 in the survival mechanism used by human cells when exposed to some of the most common forms of anticancer therapy. PMID- 9201713 TI - Mitotic spindle function in Saccharomyces cerevisiae requires a balance between different types of kinesin-related motors. AB - Two Saccharomyces cerevisiae kinesin-related motors, Cin8p and Kip1p, perform an essential role in the separation of spindle poles during spindle assembly and a major role in spindle elongation. Cin8p and Kip1p are also required to prevent an inward spindle collapse prior to anaphase. A third kinesin-related motor, Kar3p, may act antagonistically to Cin8p and Kip1p since loss of Kar3p partially suppresses the spindle collapse in cin8 kip1 mutants. We have tested the relationship between Cin8p and Kar3p by overexpressing both motors using the inducible GAL1 promoter. Overexpression of KAR3 results in a shrinkage of spindle size and a temperature-dependent inhibition of the growth of wild-type cells. Excess Kar3p has a stronger inhibitory effect on the growth of cin8 kip1 mutants and can completely block anaphase spindle elongation in these cells. In contrast, overexpression of CIN8 leads to premature spindle elongation in all cells tested. This is the first direct demonstration of antagonistic motors acting on the intact spindle and suggests that spindle length is determined by the relative activity of Kar3p-like and Cin8p/Kip1p-like motors. PMID- 9201714 TI - Saccharomyces cerevisiae genes required in the absence of the CIN8-encoded spindle motor act in functionally diverse mitotic pathways. AB - Kinesin-related Cin8p is the most important spindle-pole-separating motor in Saccharomyces cerevisiae but is not essential for cell viability. We identified 20 genes whose products are specifically required by cell deficient for Cin8p. All are associated with mitotic roles and represent at least four different functional pathways. These include genes whose products act in two spindle motor pathways that overlap in function with Cin8p, the kinesin-related Kip1p pathway and the cytoplasmic dynein pathway. In addition, genes required for mitotic spindle checkpoint function and for normal microtubule stability were recovered. Mutant alleles of eight genes caused phenotypes similar to dyn1 (encodes the dynein heavy chain), including a spindle-positioning defect. We provide evidence that the products of these genes function in concept with dynein. Among the dynein pathway gene products, we found homologues of the cytoplasmic dynein intermediate chain, the p150Glued subunit of the dynactin complex, and human LIS 1, required for normal brain development. These findings illustrate the complex cellular interactions exhibited by Cin8p, a member of a conserved spindle motor family. PMID- 9201715 TI - An unusual fibrillarin gene and protein: structure and functional implications. AB - The diploid germinal nucleus of the ciliated protozoan Tetrahymena thermophila is unusual among eukaryotes in that it encodes a single copy of the gene for rRNA allowing identification of cis-acting mutations in rDNA affecting rRNA structure, function, and processing. The generally conserved nucleolar protein fibrillarin has been characterized from a number of systems and is involved in pre-rRNA processing. We have demonstrated that Tetrahymena has fibrillarin and have analyzed the cDNA and the genomic DNA encoding this protein. The derived amino acid sequence of the N-terminal region of Tetrahymena fibrillarin shows little similarity with the generally highly conserved glycine/arginine-rich N-terminal domain of other eukaryotic fibrillarins. The remainder of the amino acid sequence of the molecule is more conserved. Polyclonal antibodies generated against the full-length Tetrahymena fibrillarin expressed in bacteria recognize a protein of M(r) approximately 32,000 in whole-cell or nucleolar preparations. Immunocytochemistry localizes fibrillarin to nucleoli in the somatic macronuclei of vegetative cells. Transformation experiments demonstrate that fibrillarin is an essential protein in Tetrahymena. The Tetrahymena fibrillarin is expressed but does not complement a NOP1 null mutation when transformed into the yeast Saccharomyces cerevisiae, indicating less functional conservation among fibrillarins than previously suggested. PMID- 9201716 TI - Genetic approach to regulated exocytosis using functional complementation in Paramecium: identification of the ND7 gene required for membrane fusion. AB - Paramecium is a unicellular organism that possesses a specialized pathway for regulated secretion that is amenable to genetic studies. Numerous mutations affecting the process have been isolated over the years, among which is a subclass blocking the terminal step of fusion of the secretory granule with the plasma membrane. We report herein the cloning by functional complementation of one such gene, ND7. The 506-amino acid polypeptide encoded by ND7 is predicted to be a type I integral membrane protein with a highly charged cytosolic domain featuring amphipathic and coiled-coil regions. This structure is compatible with the physiological data on the mutant nd7-1 suggesting that the protein is anchored in the membrane of the secretory granule and that it may interact with other proteins. This work presents the first identification by a genetic approach of a novel gene involved in regulated secretion and establishes Paramecium as a powerful model system for the genetic dissection of this process. PMID- 9201717 TI - Sequence and overexpression of GPP130/GIMPc: evidence for saturable pH-sensitive targeting of a type II early Golgi membrane protein. AB - It is thought that residents of the Golgi stack are localized by a retention mechanism that prevents their forward progress. Nevertheless, some early Golgi proteins acquire late Golgi modifications. Herein, we describe GPP130 (Golgi phosphoprotein of 130 kDa), a 130-kDa phosphorylated and glycosylated integral membrane protein localized to the cis/medial Golgi. GPP130 appears to be the human counterpart of rat Golgi integral membrane protein, cis (GIMPc), a previously identified early Golgi antigen that acquires late Golgi carbohydrate modifications. The sequence of cDNAs encoding GPP130 indicate that it is a type II membrane protein with a predicted molecular weight of 81,880 and an unusually acidic lumenal domain. On the basis of the alignment with several rod-shaped proteins and the presence of multiple predicted coiled-coil regions, GPP130 may form a flexible rod in the Golgi lumen. In contrast to the behavior of previously studied type II Golgi proteins, overexpression of GPP130 led to a pronounced accumulation in endocytotic vesicles, and endogenous GPP130 reversibly redistributed to endocytotic vesicles after chloroquine treatment. Thus, localization of GPP130 to the early Golgi involves steps that are saturable and sensitive to lumenal pH, and GPP130 contains targeting information that specifies its return to the Golgi after chloroquine washout. Given that GIMPc acquires late Golgi modifications in untreated cells, it seems likely that GPP130/GIMPc continuously cycles between the early Golgi and distal compartments and that an unidentified retrieval mechanism is important for its targeting. PMID- 9201718 TI - A novel Sec18p/NSF-dependent complex required for Golgi-to-endosome transport in yeast. AB - The vacuolar protein-sorting (VPS) pathway of Saccharomyces cerevisiae mediates localization of proteins from the trans-Golgi to the vacuole via a prevacuolar endosome compartment. Mutations in class D vacuolar protein-sorting (vps) genes affect vesicle-mediated Golgi-to-endosome transport and result in secretion of vacuolar proteins. Temperature-sensitive-for-function (tsf) and dominant negative mutations in PEP12, encoding a putative SNARE vesicle receptor on the endosome, and tsf mutations in VAC1, a gene implicated in vacuole inheritance and vacuolar protein sorting, were constructed and used to demonstrate that Pep12p and Vac1p are components of the VPS pathway. The sequence of Vac1p contains two putative zinc-binding RING motifs, a zinc finger motif, and a coiled-coil motif. Site directed mutations in the carboxyl-terminal RING motif strongly affected vacuolar protein sorting. Vac1p was found to be tightly associated with membranes as a monomer and in a large SDS-resistant complex. By using Pep12p affinity chromatography, we found that Vac1p, Vps45p (SEC1 family member), and Sec18p (yeast N-ethyl maleimide-sensitive factor, NSF) bind Pep12p. Consistent with a functional role for this complex in vacuolar protein sorting, double pep12tsfvac1tsf and pep12tsf vps45tsf mutants exhibited synthetic Vps- phenotypes, the tsf phenotype of the vac1tsf mutant was rescued by overexpression of VPS45 or PEP12, overexpression of a dominant pep12 allele in a sec18-1 strain resulted in a severe synthetic growth defect that was rescued by deletion of PEP12 or VAC1, and subcellular fractionation of vac1 delta cells revealed a striking change in the fractionation of Pep12p and Vps21p, a rab family GTPase required for vacuolar protein sorting. The functions of Pep12p, Vps45p, and Vps21p indicate that key aspects of Golgi-to-endosome trafficking are similar to other vesicle-mediated transport steps, although the role of Vac1p suggests that there are also novel components of the VPS pathway. PMID- 9201719 TI - The Cdc2 protein kinase controls Cdc10/Sct1 complex formation. AB - In the fission yeast Schizosaccharomyces pombe, the execution of Start requires the activity of the Cdc2 protein kinase and the Cdc10/Sct1 transcription complex. The loss of any of these genes leads to G1 arrest and activation of the mating pathway under appropriate conditions. We have undertaken a genetic and biochemical analysis of these genes and their protein products to elucidate the molecular mechanism that governs the regulation of Start. We demonstrate that serine-196 of Cdc10 is phosphorylated in vivo and provide evidence that suggests that phosphorylation of this residue is required for Cdc10 function. Substitution of serine-196 of Cdc10 with alanine (Cdc10 S196A) leads to inactivation of Cdc10. We show that Cdc10 S196A is incapable of associating with Sct1 to form a heteromeric complex, whereas substitution of this serine with aspartic acid (S196D) restores DNA-binding activity by allowing Cdc10 to associate with Sct1. Furthermore, we demonstrate that Cdc2 activity is required for the formation of the heteromeric Sct1/Cdc10 transcription complex and that the Cdc10 S196D mutation alleviates this requirement. We thus provide biochemical evidence to demonstrate one mechanism by which the Cdc2 protein kinase may regulate Start in the fission yeast cell cycle. PMID- 9201720 TI - Ran1 functions to control the Cdc10/Sct1 complex through Puc1. AB - We have undertaken a biochemical analysis of the regulation of the G1/S-phase transition and commitment to the cell cycle in the fission yeast Schizosaccharomyces pombe. The execution of Start requires the activity of the Cdc2 protein kinase and the Sct1/Cdc10 transcription complex. Progression through G1 also requires the Ran1 protein kinase whose inactivation leads to activation of the meiotic pathway under conditions normally inhibitory to this process. We have found that in addition to Cdc2, Sct1/Cdc10 complex formation requires Ran1. We demonstrate that the Puc1 cyclin associates with Ran1 and Cdc10 in vivo and that the Ran1 protein kinase functions to control the association between Puc1 and Cdc10. In addition, we present evidence that the phosphorylation state of Cdc10 is altered upon inactivation of Ran1. These results provide biochemical evidence that demonstrate one mechanism by which the Ran1 protein kinase serves to control cell fate through Cdc10 and Puc1. PMID- 9201721 TI - Persistent DNA damage inhibits S-phase and G2 progression, and results in apoptosis. AB - We used genetically related Chinese hamster ovary cell lines proficient or deficient in DNA repair to determine the direct role of UV-induced DNA photoproducts in inhibition of DNA replication and in induction of G2 arrest and apoptosis. UV irradiation of S-phase-synchronized cells causes delays in completion of the S-phase sometimes followed by an extended G2 arrest and apoptosis. The effects of UV irradiation during the S-phase on subsequent cell cycle progression are magnified in repair-deficient cells, indicating that these effects are initiated by persistent DNA damage and not by direct UV activation of signal transduction pathways. Moreover, among the lesions introduced by UV irradiation, persistence of (6-4) photoproducts inhibits DNA synthesis much more than persistence of cyclobutane pyrimidine dimers (which appear to be efficiently bypassed by the DNA replication apparatus). Apoptosis begins approximately 24 h after UV irradiation of S-phase-synchronized cells, occurs to a greater extent in repair-deficient cells, and correlates well with the inability to escape from an extended late S-phase-G2 arrest. We also find that nucleotide excision repair activity (including its coupling to transcription) is similar in the S-phase to what we have previously measured in G1 and G2. PMID- 9201724 TI - [A new approach to innovating selective anxiolytics: pharmacological profile of a novel 5-HT1A agonist (tandospirone)]. AB - Tandospirone (sedil) is a newly developed anxiolytic drug that has a much higher selective affinity for 5-HT1A than dopamine D2 receptors without the binding affinities with noradrenergic, dopaminergic, cholinergic and GABAergic receptors. This agent binds with 5-HT1A receptors located in both 5-HT neurons in the raphe nucleus and other postsynaptic neurons to induce hyperpolarization of the neurons by opening the K+ channels to eventually inhibit the target neuronal activities. With repeated administrations of tandospirone, a decrease in 5-HT2A receptor population was observed. Behavioral studies in experimental animals have demonstrated that tandospirone inhibits conflict in Vogel methods, aggressive behavior and muricide in manners similar to those of diazepam. In addition, tandospirone showed antistress effects in experimental models and antidepressive effects in forced swimming tests. Unlike diazepam, tandospirone does not produce sedative, sleep-inducing, anticonvulsant, nor muscle relaxant effects at doses effective for conflict tests. Drug dependance, one of the serious problems with bezodiazepine, is not observed with repeated treatment of tandospirone in rats and monkeys. Furthermore, tandospirone has been reported to show a significantly more superior or equipotent effect to diazepam in controlling autonomic disturbances, psychiatric cardiovascular and vegetative syndromes as well as neurosis in double blind clinical studies. These effects are probably due to the selective action of tandospirone on 5-HT1A receptors in the limbic system to eventuate anxiolytic and antidepressant effects. A decrease in 5-HT2A receptor population with repeated treatment of tandospirone may have contributed to the antidepressive effect. Furthermore, 5-HT1A receptors relatively, selectively distributed in the limbic system are not involved in sedation, sleep or muscle relaxation. Such unwanted effects of benzodiazepines are thus not observed with tandospirone treatment. PMID- 9201725 TI - [Context-dependent sensitization: reconsideration and a hypothesis]. AB - The repeated administration of amphetamine-like psychostimulants results in an augmentation of their behavioral responses, a phenomenon known as behavioral sensitization. One important factor associated with the process of behavioral sensitization is environmental influence. It has been reported that, when drug administration is paired with a particular environment, sensitization is observed only in that particular environment. This phenomenon has been known as context dependent sensitization. However, considering recent reports and our own studies, the classical concept of context-dependent sensitization may not be satisfactory. We propose an alternative hypothesis. Psychostimulants are known to induce different behaviors in different environments. We believe that the repeated administration of a psychostimulant in different environments results in the augmentation of different behaviors. For instance, rats treated with a stimulant in a small cage did not locomote but were observed to sniff and rear. After repeated treatment, they showed sensitization not in locomotion but in stereotyped behaviors such as sniffing and rearing. This suggests that environment does not facilitate the development of sensitization, but rather modifies the pattern and character of a stimulant-induced behavior in the sensitized animals. In this paper, we briefly review various literature and present our hypothesis. PMID- 9201723 TI - Palmitoylation of p59fyn is reversible and sufficient for plasma membrane association. AB - Members of the Src family of protein tyrosine kinases are localized to subspecialized regions of the plasma membrane. Herein we show that the N-terminal SH4 region of the Src family member p59fyn (Fyn) is both necessary and sufficient for targeting of Fyn and heterologous proteins to the plasma membrane and detergent-insoluble subdomains. Attachment of the first 16 amino acids of Fyn to a normally cytosolic protein, beta-galactosidase, resulted in distinct plasma membrane localization of the chimeric protein. Mutation of the palmitoylation site (cysteine-3) within Fyn16-beta-galactosidase or wild-type Fyn abrogated plasma membrane localization, resulting in redistribution of the mutant proteins into intracellular membranes. Substitution of the SH4 motif within Fyn with heterologous sequences from other palmitoylated proteins (G alpha o and GAP43) revealed that the presence of palmitate is sufficient to direct plasma membrane localization independent of surrounding amino acid sequences and myristate. Palmitoylated Fyn chimeras were also enriched in the Triton X-100-resistant matrix, whereas nonpalmitoylated forms of these proteins were detected in the detergent-soluble fraction. The palmitate moiety on Fyn exhibited a half-life of 1.5-2 h. In contrast, the half-life of the polypeptide backbone was 8 h, indicating that palmitoylation is a reversible modification. These studies establish that the palmitoylated SH4 sequence of Fyn can be used to specifically target proteins to the plasma membrane in a reversible manner. PMID- 9201722 TI - RCC1 and nuclear organization. AB - We have examined the effect of RCC1 function on the nuclear organization of pre mRNA splicing factors and poly(A)+ RNA in the tsBN2 cells, a RCC1 temperature sensitive mutant cell line. We have found that at 4-6 h after shifting cells from the permissive temperature (32.5 degrees C) to the restrictive temperature (39.5 degrees C), both small nuclear ribonucleoprotein particles and a general splicing factor SC35 reorganized into 4-10 large round clusters in the nucleus, as compared with the typical speckled distribution seen in cells at the permissive temperature. In situ hybridization to poly(A)+ RNA resulted in a similar pattern. Examination by double labeling demonstrated that the redistribution of splicing factors coincides with that of poly(A)+ RNA. Such changes in the nuclear organization of splicing factors and poly(A)+ RNA were not the result of the temperature shift or of chromatin condensation. Cellular transcription was not significantly altered in these cells and extracts made from both the permissive and restrictive temperature were splicing competent. Electron microscopic examination demonstrated that the large clusters containing both splicing factors and poly(A)+ RNA were fused interchromatin granule clusters. In addition, small electron-dense dot-like structures measuring approximately 80 nm in diameter were also observed, most of which are accumulated in enlarged interchromatin granule clusters in the nucleoplasm of RCC1- cells. In spite of the significant changes observed in the nucleoplasm, relatively little alteration was observed in nucleolar structure by both light and electron microscopic examination. The above observations suggest that the RCC1 protein directly or indirectly regulates the organization of splicing components and poly(A)+ RNA in the cell nucleus and that RCC1 may play a role in nuclear organization. PMID- 9201726 TI - 5-Hydroxytryptamine1B and alpha 2 adrenaline receptors in the brain of rats prenatally treated with methylazoxymethanol. AB - Methylazoxymethanol (MAM)-induced cerebral hypoplasia resulted in a significant increase in concentrations of 5-hydroxytryptamine (5-HT, serotonin) and norepinephrine in the frontal cortex, suggesting that these monoaminergic neurons were compressed due to smaller brain volumes. The serotonergic and noradrenergic presynaptic autoreceptors in rat brain are thought to be 5-HT1B receptors and alpha 2-adrenoceptors, respectively. If so, prenatal MAM treatment should increase the density of 5-HT1B and adrenaline alpha 2 receptors in the brain via the compression of noradrenergic and serotonergic axon terminals in the brains of rats with MAM-induced microencephaly. However, neither the densities nor the affinities of 5-HT1B and adrenaline alpha 2 receptors were changed in the MAM rats, suggesting that these presynaptic autoreceptors comprise only a small percentage of the total receptor population. Most 5-HT1B and adrenaline alpha 2 receptors were localized post-synaptically. PMID- 9201728 TI - Cholinergic and GABAergic interneurons in the striatum. AB - In the striatum, interneurons have not been as well characterized physiologically as the spiny projection cells. We found that the neostriatal interneurons can be divided at least into three classes by physiological, chemical and morphological criteria. The first was FS cells (fast-spiking cells) which fired very short duration action potentials at constant spike frequency during depolarizing pulses, were immunoreactive for parvalbumin (calcium-binding protein), and had axons with very dense collateralization within or near their dendritic fields. Another class was identified as those which fired low-threshold spikes (LTS cells) from hyperpolarized potentials, were positive for somatostatin and nitric oxide synthase (NOS), and had the largest axonal fields. The other class of interneurons had longer-lasting afterhyperpolarizations (LA cells), were positive for choline acetyltransferase, and were mostly large aspiny cells. Glutamic acid decarboxylase (GAD67) or GABA immunoreactivity was detected at the somata or terminals of parvalbumin FS cells and somatostatin/NOS LTS cells, but not of cholinergic LA cells. Substance P, probably released from the collaterals of cells projecting to the substantia nigra, excited LA cells and LTS cells, but not FS cells. These results suggest that the striatum has at least one type of cholinergic and two types of GABAergic interneurons which are different in physiological, chemical and pharmacological characteristics. PMID- 9201727 TI - [A study of the effects of antidepressants on the GABAA receptor and its complex based on the drug actions on the power-spectral changes of rat hippocampal EEG induced by GABA antagonists and inverse agonists]. AB - To analyze the effects of antidepressants on the GABAA receptor, we investigated how the chronic administration of antidepressants (10 mg/kg twice a day for three or seven days, ip) influenced the power-spectral changes induced by pentylenetetrazol (PTZ; a GABA antagonist; 27.5 mg/kg) or beta-carboline-3 carboxylic-acid-methylester (beta-CCM; an inverse agonist; 1 mg/kg) on rat hippocampal EEGs. PTZ and beta-CCM are known to inhibit the chloride ionophore and benzodiazepine receptor (GABAA receptor complex), respectively. After the ip injection of both compounds, the EEG power under 12 Hz increased to about five times that before injection. Between the rats that did not receive any antidepressants and all those injected with the drugs for 3 days or treated with desipramine (DMI) for 7 days, there were no apparent changes in the effect of PTZ or beta-CCM. However, in the rats treated with imipramine, fluoxetine or trazodone for 7 days, the increase in power after the injection of PTZ or beta CCM was apparently suppressed. In these rats, the power values were less than three times those before the dosing of PTZ or beta-CCM. DMI is known to inhibit the re-uptake of norepinephrine (NE), while the other three antidepressants inhibit that of serotonin (5-HT). Trazodone is also reported to block the 5-HT2 sites. These observations might indicate that the chronic administration of antidepressants prompted the function of the GABAA receptor complex. Moreover, it is also suggested that, to that action, the effect of antidepressants on the 5-HT system or interaction between the 5-HT system and GABA receptors might play some role. PMID- 9201729 TI - DA D2 receptors in the ventral striatum: multiple effects or receptor subtypes? AB - This short review summarizes work from the authors' laboratory regarding electrophysiological roles for two members of the dopamine (DA) D2 receptor family, the D2 and D2 receptors, and their interactions with DA D1 receptors, within the ventral striatum. The inhibitory effects of D2 receptor class agonists on ventral striatal neurons appears to be mediated by D2 as opposed to D2 receptors. This effect requires simultaneous stimulation of the D1 (D1A) receptor. The inhibitory effects of D1 class receptor agonists may involve both the D1 receptor as well as a D1-like receptor not linked to adenylyl cyclase. This effect does not require co-stimulation of a D2 class receptor. The ability of DA to enhance the excitatory effects of glutamate on ventral striatal neurons requires stimulation of both D1 and D2 class receptors. Stimulation of D2 class receptors can either increase or decrease whole cell sodium currents; the particular D2 receptor subtypes responsible for these effects have yet to be established. PMID- 9201730 TI - D2 receptor activation in distinct striatal neurons in comparison with D3 receptors. AB - The role of D2/D3 receptors in striatum was electrophysiologically examined in vitro in chloralose-anesthetized rats. In addition, in vitro patch clamp method with rat brain slices was followed. Stimulations of the substantia nigra pars compacta (SN) in vivo elicited spike generation which was inhibited by microiontophoretically applied domperidone, a D2 antagonist. These domperidone sensitive neurons were activated by microiontophoretic application of D2 agonists such as talipexole, quinpirole and bromocriptine as well as the D2 agonist, 7-OH DPAT. They were also excited by either intravenous injection of bromocriptine or talipexole in a dose-dependent manner. Furthermore, the SN-induced increases in neuronal firing were blocked during microiontophoretic application of domperidone. In patch clamp whole-cell recording large-sized cells, identified visually under Ramanosky microscope, were depolarized with repetitive firing on bath application of talipexole and 7-OH-DPAT at a current clamp mode. Talipexole induced depolarization in the large-sized cell was similarly observed in the presence of TTX and high Mg2+ in Ca(2+)-free physiological solution. In contrast, the medium-sized cells were hyperpolarized on bath application of talipexole without being affected by 7-OH-DPAT. These findings suggest that the large-sized cells, which were presumably cholinergic interneurons, are activated by dopamine derived from the SN via D2 and/or D3 receptors, while the medium-sized cells are inhibited by dopamine via D2 receptors. PMID- 9201731 TI - Role of dopamine receptors in the short- and long-term regulation of corticostriatal transmission. AB - Several studies have tried to clarify the role of different dopamine (DA) receptors in the control of membrane excitability of striatal neurons. Activation of DA receptors influences both synaptic and intrinsic membrane properties of striatal neurons. More recently it has been reported that endogenous DA plays an important role in the expression of striatal synaptic plasticity. In this review we will try to summarize and discuss the available data concerning the possible impact of the functional role of D1 and D2 receptor activation on the short- and long-term modulation of the excitatory glutamatergic corticostriatal transmission. Moreover, we will also describe the function of the striatum in the integration of glutamatergic and DAergic inputs to produce long-term changes of synaptic efficacy: long-term depression (LTD) and long-term potentiation (LTP). PMID- 9201732 TI - State-dependent regulation of neuronal excitability by dopamine. AB - Since the discovery that the loss of the dopaminergic innervation of the striatum resulted in Parkinson's disease, physiologists have attempted to understand the role of dopamine on striatal activity. Hypotheses relying upon concepts derived from studies of fast synaptic transmission have consistently failed to explain the actions of dopamine or other receptors coupled to G-proteins which modulate the properties of voltage-dependent ionic conductances responsible for synaptic integration and spike activity. Recently, patch clamp studies have revealed that in medium spiny striatal neurons dopamine D1-class receptors modulate voltage dependent Na+, K+ and Ca2+ channels. From a consideration of the biophysical properties of these channels and the state transitions that medium spiny neurons undergo while responding to cortical input, a novel picture of dopamine's actions is beginning to emerge. Our results and those of others suggest that D2-class receptors serve to make the transition to the depolarized 'upstate' from the hyperpolarized 'downstate' more probable in response to cortical input. But, once the transition has occurred, the alteration in excitability should be short-lived unless the neuron has recently been active. This state-dependent modulation provides a mechanism by which dopamine could shape global striatal activity governing the execution of motor behaviors. PMID- 9201733 TI - Do D2-like dopamine receptors mediate neuronal excitation or inhibition: some functional-behavioural implications. AB - Few controversies in contemporary neuroscience have endured as long as that concerning the immediate neurophysiological effect(s) of 'D2-like' dopamine receptor activation. While the issue is of fundamental importance, it could be argued that the matter is more abstract and of less consequence than clarifying receptor-mediated actions at other, more functional levels from second messenger through neurochemistry to behaviour. Yet it is not possible to define fully these processes without insight into the immediate neurophysiological action. This necessity has been accentuated by a number of recent developments, including (i) recognition of multiple, complex forms of functional interactions between 'D2 like' and 'D1-like' receptor families which ultimately regulate so many forms of behaviour and (ii) the ongoing debate as to the extent to which members of 'D1 like' and 'D2-like' receptor families might be co-localised on the same neuronal membrane to subserve such interactions. Full specification of these phenomena requires a clear understanding of the relevant neurophysiological events. PMID- 9201734 TI - Patients can be pain-free while undergoing implanted port assessment. PMID- 9201735 TI - A descriptive study of the role of the oncology nurse practitioner. AB - PURPOSE/OBJECTIVES: To describe the characteristics and activities of nurse practitioners (NP) with a focus in oncology. DESIGN: Descriptive. SAMPLE: 129 NPs employed in an oncology setting who completed on NP program and were functioning in the NP role. METHODS: Subjects completed an eight-page, self-administered questionnaire comprised of fixed-choice and open-ended questions. MAIN RESEARCH VARIABLES: Demographics, employment settings, populations served, advanced practice subroles, clinical functions, practice privileges, reimbursement issues, job descriptions, performance appraisals, job satisfaction, and facilitators/barriers to role implementation. FINDINGS: The majority of oncology NPs (ONPs) were located in the eastern United States in university-affiliated hospitals. The most common patient population served by the respondents was adults in the medical oncology outpatient setting. More than three-quarters of the respondents worked from protocols, almost two-thirds performed procedures traditionally performed by physicians, and approximately half had prescriptive authority. Few NP respondents reported that they obtained direct reimbursement for their services from third-party payors. Physicians were cited as the most facilitative of the NP role, and administrators were cited as the most frequent barrier. The vast majority of the respondents were satisfied with their roles. IMPLICATIONS FOR NURSING PRACTICE: The NP role in oncology is established and expanding. The scope of practice and more detailed characterization of the role is an area for future research. Data on the effectiveness of ONPs, particularly regarding cost-effectiveness, quality of care, and patient satisfaction, are needed to maintain their viability within the healthcare system. PMID- 9201736 TI - Getting a Pap smear: focus group responses of African American and Latina women. AB - PURPOSE/OBJECTIVES: To identify barriers and facilitating factors associated with Papanicolaou (Pap) smear use among African American and Latina women. DESIGN: Descriptive, exploratory. SAMPLE AND SETTING: Fifty-two African American and Latina women recruited from health and social service agencies. METHODS: Guided by the Theory of Planned Behavior, focus group interviewing and an open-ended questionnaire were used to identify cancer beliefs and salient behavioral beliefs, referents, and control beliefs about Pap smears. FINDINGS: False negative results, financial burden, and the role of the physician were the most salient beliefs for African American women. For Latinas, embarrassment, use of a cold or unclean speculum, and discomfort were the most salient beliefs. Salient referents for African American and Latina women were physicians and male partners; access to Pap smear screening and financial assistance were their frequently held control beliefs. Beliefs regarding cancer fatalism were consistent with reports of national samples of African American and Latino adults. CONCLUSIONS: Cancer beliefs among African Americans and Latinas have not changed significantly in the past 10-15 years, despite increased national cancer education and screening advertisements. Moreover, as issues regarding Pap smear use remain the same, new concerns develop. Specifically, beliefs about speculums, perceptions of discomfort, perceptions of disapproval of physicians and male partners, fear as a motivator and inhibitor, and lack of information about cervical cancer and Pap smear screening present new concerns for these women. IMPLICATIONS FOR NURSING PRACTICE: The findings provided data needed to develop a questionnaire that will be used to test Pap smear screening intentions among African American and Latina women. PMID- 9201737 TI - One school's experience with the development of an oncology nurse practitioner curriculum. AB - PURPOSE/OBJECTIVES: To describe the foundational work and implementation of a nurse practitioner (NP) curriculum geared toward oncology nurses. The study is selective (not comprehensive) and reflective of one school's experience. DATA SOURCE: Journal articles, curriculum guidelines, anecdotal experience, and interviews. DATA SYNTHESIS: The NP is used more frequently in oncology, both as a clinician and for other aspects of advanced practice nursing. NPs must be prepared to fulfill graduate criteria as outlined by definitive sources for curriculum development. CONCLUSIONS: Schools must work with employers, graduates, and patients in conducting outcome evaluations to measure safety issues and role effectiveness of oncology NPs (ONPs), as well as fulfillment of all aspects of the advanced nursing practice role. IMPLICATIONS FOR NURSING PRACTICE: If healthcare employers continue to rely heavily on the use of ONPs, schools of nursing must be prepared to graduate safe clinicians, experts in oncology, and advanced practice nurses, all combined into one graduate. This difficult task requires evaluation of current practices. PMID- 9201738 TI - Intravenous drug compatibility: a challenge for the oncology nurse. AB - PURPOSE/OBJECTIVES: To examine the cause(s) and types of IV incompatibilities that can occur in the oncology population, to present information on commonly used oncologic medications, to identify resources available to nurses, and to provide specific interventions designed to prevent incompatibilities in various oncology settings. DATA SOURCES: Published articles, nursing and pharmacology textbooks. DATA SYNTHESIS: Many common oncologic medications are not compatible. A number of factors, including pH, drug additives, and cosolvents, may cause these incompatibilities. Incompatible reactions may be visually apparent or may occur on a molecular level. CONCLUSIONS: A basic understanding of incompatibilities can minimize their occurrence. A variety of resources and interventions are available. IMPLICATIONS FOR NURSING PRACTICE: Incompatibilities can occur easily in a variety of oncology settings and can result in detrimental medical and financial outcomes. Nurses play an important role in preventing incompatibilities. They must be familiar with the medications they are administering and have access to compatibility information and resources. PMID- 9201739 TI - Women's experiences of lymphedema. AB - PURPOSE/OBJECTIVES: To explore women's experiences of lymphedema. DESIGN: Qualitative descriptive. SETTING: An urban community in the midwestern United States. SAMPLE: Ten women who experienced lymphedema after breast cancer treatment and who had (a) completed their treatment for stage I or stage II breast cancer at least one year before the study, (b) experienced an onset of lymphedema at least two months after surgery, (c) no current evidence of cancer disease or cancer recurrence, (d) lymphedema not caused by cancer in the brachial plexus, and (e) no history of hospitalization for alcoholism, substance abuse, or psychiatric conditions. The women ranged in age from 36-75 years. Mean survival time was seven years, and the mean time since onset of lymphedema was four years. METHODS: Two in-depth interviews per participant. PATIENTS: Most women were able to continue living their normal lives. Some women experienced depression, anxiety, and impairments related to their intimate, work, and social relationships. Physicians' limited knowledge about lymphedema, conflicting treatment information, and the limited number of available treatment centers caused distress for the participants. Their experiences can be categorized into three predominant themes: Abandonment by Medicine, Concealing the imperfect image, and Living the Interrupted Life. CONCLUSIONS: Research efforts to expand the knowledge base regarding the epidemiology, prevention, and treatment of lymphedema are needed. Also needed is research that explores the impact of lymphedema on quality of life and functional ability as well as the psychosocial impact of lymphedema on body image, self esteem, and social support. IMPLICATIONS FOR NURSING PRACTICE: Care providers and breast cancer survivors should be educated about the prevention and treatment of lympedema. Nurses should refer patients to advocacy hot lines and support groups for information and support when appropriate. Women with lymphedema should be assessed periodically for psychosocial distress and referred for care as needed. PMID- 9201740 TI - Incentives and barriers to exercise in women with a history of breast cancer. AB - PURPOSE/OBJECTIVES: To develop and test an instrument to identity the most important incentives and barriers to exercise in women with breast cancer. DESIGN: Descriptive, quantitative, retrospective. SETTING: Ambulatory, national. SAMPLE: Phase I-11 Caucasian women, with a mean age of 43 years, who were treated for breast cancer within the past five years; Phase II-64 primarily Caucasian women, with a mean age of 47 years, who were treated for breast cancer within the past 10 years. METHODS: Content analysis of structured interviews, questionnaire. FINDINGS: The mean score on the decisional balance index (DBI), an overall exercise weight, was a low +19. The range of scores was from a low -26 to a moderate +57 on a scale from -72 to +72. The DBI accurately predicted 88% of the distribution of women who exercised regularly and those who did not. The most important incentives for exercise in a sample of women with a history of breast cancer were expectation of benefit and sense of responsibility. The most important barriers were lack of time and inertia. CONCLUSIONS: Replication is needed with larger, more diverse samples, but the Incentive and Barriers to Exercise Scale (IBES) potentially can screen women with breast cancer with the greatest need for exercise interventions. An individualized plan to promote incentives and reduce barriers should be incorporated into interventions. IMPLICATIONS FOR NURSING PRACTICE: Nurses' proactive support for and education about exercise is needed. PMID- 9201741 TI - Expectations and experiences of patients with cancer participating in phase I clinical trials. AB - PURPOSE/OBJECTIVES: To describe the expectations and experiences of patients entering phase I clinical trials. DESIGN: Descriptive, exploratory, prospective. SETTING: A large military medical teaching center. SAMPLE: Thirty-seven adult patients completed the entry and exit interviews. Subjects had a good performance status, were middle aged, and had common tumor types. METHODS: Interviews using structured entry and exit questionnaires. MAIN RESEARCH VARIABLES: Expectations and experiences of patients in phase I clinical trials. FINDINGS: Patients expected slightly increased support from family members and received more support than expected. Patients' expectations for tumor response and increased communication with their physician were not met. Patients expected symptoms such as fatigue, nausea and vomiting, and weight loss to improve during therapy, yet their expectations were not met. CONCLUSIONS: One theme that emerged from the data was hope/optimism. An issue that needs further exploration is the extent to which patients accurately understand information in the consent form. Findings also support the importance of communication between the patient and family members and the healthcare team. IMPLICATIONS FOR NURSING PRACTICE: Oncology nurses can mediate the flow of information between physicians and patients. Oncology nurses can and should assess the patient's level of understanding of the clinical trial, the consent form, and potential side effects at the time of entry into the trial and intermittently during the course of therapy. Nurses must allow patients with cancer who are undergoing investigational therapy to maintain a level of hope, while realistically counseling them about their progress during phase I trial participation. PMID- 9201742 TI - A comparison of the level of hope in patients with newly diagnosed and recurrent cancer. AB - PURPOSE/OBJECTIVES: To compare levels of hope in patients with newly diagnosed and recurrent cancer. DESIGN: Descriptive study. SETTING: Three oncology practices in two urban areas of the southern United States. SAMPLE: Convenience sample of 20 newly diagnosed patients with cancer and 16 patients with recurrent cancer (mean age = 56 years). The majority of the patients were Caucasian, female, and married; had a high school degree; and had a religious affiliation. METHODS: Subjects completed the Herth Hope Scale and answered the open-ended question "What gives you the most hope at the present time?" Analysis included descriptive statistics (i.e., frequency, means, standard deviations, percents), t tests, Chi-square, and analysis of variance. MAIN RESEARCH VARIABLES: Level of hope each subject had in relation to the stage of the cancer at the time of diagnosis. FINDINGS: Contrary to expectations, patients with newly diagnosed and recurrent cancer did not differ in regard to their level of hope. However, significant differences were found related to the type of hope utilized. Married patients and male patients experienced higher levels of hope. Recurrent themes in response to the open-ended question were family support, nonfamily support, faith, outlook, and health professionals/care. CONCLUSIONS: Patients with newly diagnosed cancer use their treatment and nurses, physicians, and other healthcare professionals as sources of hope and support. Patients with recurrent cancer reported drawing hope from faith. IMPLICATIONS AND NURSING PRACTICE: Heightened awareness of the patient-healthcare professional relationship will enable healthcare professionals to provide care that is more sensitive to one congruent with patients' needs. Healthcare professionals need to assess the meaning of faith for each individual patient and offer services to foster this source of hope. PMID- 9201743 TI - A comprehensive interdisciplinary chemotherapy teaching documentation flowsheet. AB - PURPOSE/OBJECTIVES: To describe the development and implementation of one approach to standardize and document interdisciplinary chemotherapy education for patients and families. DATA SOURCES: Cancer center interdisciplinary team. Oncology Nursing Society standards of care, clinical experience, and published literature. DATA SYNTHESIS: Because chart reviews of patient records demonstrated inconsistent chemotherapy education, a comprehensive chemotherapy curriculum pain was designed as a template for patient education. A flowsheet was developed to document use of the patient curriculum as well as other chemotherapy-related education materials. Patient chemotherapy curriculum included information regarding cancer and its treatment, adverse effects, and self-care measures and material about psychosocial and spiritual care. CONCLUSIONS: A standardized approach dramatically improved chemotherapy-related patient education as well as interdisciplinary documentation of patient education. The curriculum and the flowsheet are interdisciplinary and consistent with the patient and family education standards required by the Joint Commission on Accreditation of Health Care Organizations. IMPLICATIONS FOR NURSING PRACTICE: The combination of a patient chemotherapy curriculum and a documentation flowsheet saves time and standardizes content. Patient information is comprehensive and consistent, yet open to individual interpretation. Use of the flowsheet also has increased interdisciplinary collaboration, and the standardized curriculum has decreased redundancy between providers because all members of the interdisciplinary team know what is required and what information has been taught by whom. PMID- 9201744 TI - Jack bean (Canavalia ensiformis): nutrition related aspects and needed nutrition research. AB - The nutritional characteristics and food potentials of jack bean (Canavalia ensiformis) have been reviewed. The bean is a good sources of protein, 23% to 34%, and carbohydrate 55%. It is also a good source of Ca, Zn, P, Mg, Cu and Ni. Jack bean protein is adequate in most essential amino acids with the exception of methionine and cystine which may be nutritionally limiting. Antinutritional and toxic factors including trypsin inhibitors, hemagglutinins, cyanogen glucosides, oligosaccharides and others are present in jack bean. Properly processed jack bean could be used to prepare some of the popular dishes made from cowpea, peanut, pigeon pea and soybean. Industrial products such as protein concentrates and isolates, starch, flakes, grits and flours can be produced from the bean. Further research is needed to identify varieties with high protein and nutritional quality. Development of new highly nutritious food products based on whole or processed jack bean should increase production and expand use. PMID- 9201745 TI - Physicochemical characteristics of five date fruit cultivars grown in the United Arab Emirates. AB - The physical measurements and chemical analyses of date fruits of five cultivars grown in the United Arab Emirates were measured in this study. Due to differences in seed weight, the flesh accounted for 83-92% of the total fruit. At the tamr stage, the absence of sucrose and the presence of higher concentrations of reducing sugars, especially fructose and glucose, characterized these cultivars as the soft type. On maturation from the kimri to the tamr stage, the sugar content had increased, but other constituents like moisture, crude protein, crude fat, ash, crude fiber, tannins, and pectin had decreased. PMID- 9201746 TI - Microspectrophotometric evaluation of digestibility of pollen grains. AB - Digestibility of pollen grains of poppy (Papaver rhoeas) and hazelnut (Corylus avellana) subjected to a human-like in vitro digestion with pancreatic enzymes was evaluated. Pollens showed different types of walls. Digestibility was determined for total protein and insoluble carbohydrate contents by means of a new application of microspectrophotometry. Results demonstrated that pollen grains of both species were only partly digested; after 24 h treatment, only 26% of carbohydrates and 48% of proteins were digested in poppy and only 3% and 59% in hazelnut. This is probably due to the difficulty of enzymes to penetrate the intine of pollen grains. The degree of digestion of insoluble carbohydrates varied in the studied species according to their chemical nature and their storage sites. PMID- 9201748 TI - Cotyledon thermal behavior and pectic solubility as related to cooking quality in common beans. AB - The characteristic of proteins, starch and pectic substances in cotyledons of two bean cultivars varying in cooking time were determined to investigate their possible contribution to bean cooking quality. Both cultivars showed the same enthalpies of starch gelatinization but different protein denaturation enthalpies. The proportion of hot water soluble pectins was higher in Michigan, the cultivar with the lower cooking time, than in Ojo de Cabra, the cultivar with the higher cooking time. These results were not due to differences in pectin methylation or in the ratio of monovalent to divalent cations in the tissue, suggesting that in fresh beans the beta-elimination reaction is not the sole or predominant route of thermal pectin degradation. Overall, this study indicates that varietal differences in bean cooking quality may be reflections of the rate of pectin loss during soaking/heating and that the thermal properties of starch and protein fractions seem to have a minor contribution. Researchers involved in this study propose that in fresh beans, the thermal pectin loss results from a two step mechanism: pectin enzymic breakdown during the bean soaking followed by thermal solubilization rather than beta-elimination during the bean heating. PMID- 9201747 TI - Functional suitability of commercially milled rice bran in India for use in different food products. AB - The effect of blending of commercially available full fat and defatted rice brans in India from modern multistage rice mills with parboiling/stabilizing facilities in different food products in comparison to those obtained from laboratory milling of rice is reported. Bread volume and cookie spread decreased but muffin volume increased with the addition of different types of bran to wheat flour, however, the cookie spread factor was not affected by addition of full fat rice bran. The yields of the extrudate were increased by the blending of full fat rice bran but were decreased by the addition of defatted rice bran. Rice brans could be added to different food products to the extent of 5-10%. However, the full fat rice bran could not be used for production of extruded snack food. PMID- 9201750 TI - Enzymatic treatment to enhance carotenoid content in dehydrated marigold flower meal. AB - The effect of enzymatic treatments using a commercial enzyme (Econase-cep at pH 5.0 and 0.1% w/w concentration) at different levels of dehydrated marigold meal (5, 10, 15 and 20% dry weight), to enhance carotenoid extraction, was evaluated. The AOAC method consisting of a simultaneous extraction and saponification by using hexane-ethanol-acetone-toluene (10:6:7:7 v/v) and metanolic KOH (40% w/v) was used to evaluate the carotenoid content. The measurement of carotenoids was also carried out in samples in which the water soluble compounds were previously eliminated (AOAC-H2O). Total carotenoids ranged from 11.4 to 17.4 g/kg and 18 to 24.7 g/kg of control and treated marigold meal, respectively. Highest amount of carotenoids were noted when 5% level of treated meal was used. PMID- 9201749 TI - Amino acid composition, available lysine content and in vitro protein digestibility of selected tropical crop seeds. AB - As the search for alternative sources of food to alleviate hunger continues, this study was undertaken to determine nitrogen and amino acid content, chemical score, protein digestibility corrected amino acid score, available lysine and in vitro digestibility of 8 lesser known, wild tropical seeds, gathered in Nigeria. Results were contrasted with a tropical soybean variety (Glycine max, TGX 1660 15F). The investigated seeds were Millettia thonningii, Gliricidia sepium, Lonchocarpus sericeus, Albizia zygia, Daneillia ogea and Afzella bella from the family of Leguminosae, Diospyros mespiliformis (Ebenaceae) and Entandrophragma angolense (Meliaceae). The crude protein content, based on nitrogen determination, was found to be lower in the wild seeds compared to soybean, which was partly due to the relatively high content of non-protein nitrogen. With reference to amino acid requirement and digestibility in most seed samples, lysine, followed by sulphur amino acids and threonine, were the limiting amino acids. It was concluded, that these less familiar wild seed plants may be used as valuable food or feed complements. However, further investigation is necessary to elucidate potential toxic and antinutritional factors. PMID- 9201752 TI - Psychological stress in middle-aged and aged executives. AB - Neurotic symptoms were considered to be present in 21.9% of the 210 employees of a company following data analysis from a General Health Questionnaire (GHQ, 60 question items). The results in middle-aged and aged executives were compared. The total GHQ score was > or = 17 in 18/55 executives aged 40 years or above, but in only 9/60 non-executives controls was a significant difference observed (P < 0.05) between executives and non-executives. The scores for three subordinate scales, namely somatic symptoms, anxiety and insomnia, and social dysfunction, were higher in the executives than in the non-executives and showed a significant difference (P < 0.05) in the anxiety and insomnia scale. These results suggest that middle-aged and aged executives are more likely to have neurotic symptoms and more frequently to have anxiety and insomnia than their controls. We would like to apply this method to aid in the prevention, early detection and early management of stress. PMID- 9201751 TI - Glycemic index of grain amaranth, wheat and rice in NIDDM subjects. AB - Glycemic index of grain amaranth, wheat and rice preparations was studied in non insulin dependent diabetic subjects. Diets containing 50 g carbohydrate equivalent were given and post-prandial blood glucose estimated at different intervals. Glycemic index calculated for different experimental diets showed that GI of amaranth-wheat composite flour diet (25:75) was the least (65.6%) followed by wheat diet (65.7%), rice diet (69.2%), amaranth-wheat flour 50:50 (75.5%), and popped amaranth in milk (97.3%). Therefore 25:75 combination of amaranth and wheat, wheat and rice can be considered low GI food, 50:50 grain amaranth and wheat medium GI food and popped amaranth and milk combination high GI food. PMID- 9201753 TI - The relationship of DSM-III-R personality disorder to clinical variables in patients with major depression: possible difference between personality disorder clusters. AB - The relationship of DSM-III-R personality disorder (PD) to demographic and clinical variables was investigated based on 96 consecutive outpatients with major depression. No significant difference in the variables was found between those with and those without PD. Those with PD from each cluster were compared with those without PD in terms of the variables. In these comparisons many relationships of PD to the variables were found, and these relationships were different between the three PD clusters detailed in DSM-III-R. Patients with cluster B PD demonstrated a prominent uniqueness in his/her relationship to the variables. This uniqueness was similar to what had been reported previously with regard to patients with PD. There was no significant difference in the variables between those with cluster C PD and those without PD. Those with cluster A PD may have a negative family history of affective disorders. PMID- 9201754 TI - Adolescents with developmental psychopathology in adulthood. AB - We conducted a comparative study of the outcomes at 7 years and at 17 years after initial diagnosis of 77 cases of adolescent developmental psychopathology. The results suggest that the prognosis of adolescent developmental psychopathology could be derived at 5-6 years after commencement of medical treatment. The parent child relationships during infancy to adolescence, considered to affect the long term prognosis of adolescent developmental psychopathology, was studied. It is believed that the emotional bond between parents and children in early to middle childhood are vital which seems to affect their relationships during pre adolescence to early adolescence and to the development of adolescent turmoil. It can be considered vital that the development of the infantile provocative state during preadolescence to early adolescence should progress to a more mature ability to negotiate and which forms the core of adolescent turmoil. The function of the latter will bring better results to adolescent developmental psychopathologies. PMID- 9201755 TI - Acute polymorphic psychosis in adults with mild intellectual deficits. AB - This paper describes two representative cases of acute polymorphic psychotic disorder (APPD) defined in the International Classification of Diseases (ICD)-10 in adults with mild intellectual deficits. The patients showed mild intellectual impairment but had worked diligently for many years without their impairment being noticed by others. Some displayed maladjustments in their childhood and were classified as having intellectual disabilities. With their low self-esteem, precipitating events forced the patients to act inappropriately and, as a result, they became psychotic. Although APPD is supposed to occur without causative stress, an intellectual limitation combined with a certain personality can precipitate such a psychosis. It is noted that APPD might mask intellectual deficit and their precipitating factors and that a relapse could be prevented by advising a member of the patient's family. PMID- 9201756 TI - Alcohol dementia and alcohol delirium in aged alcoholics. AB - In the present study, 126 alcoholics aged 60 years or older were compared with 104 alcoholics aged 35-45 years. No dementia was found in the younger group, whereas 62.7% of the aged patients had dementia; the dementia being irreversible in 32.9% of such patients. Cases of so-called alcohol dementia excluding organic brain diseases accounted for 42.1%. The percentage of aged alcoholics having dementia increased with age, being far beyond the frequency of senile dementia in the general aged. Among various physical complications, hepatic injury and myocardiopathy were more frequent in the aged alcoholics than in general aged people, suggesting that hypertension, myocardiopathy and hepatic injury underlie the manifestation of dementia. There was no case of dementia attributable to the direct effect of alcohol distinctly exceeding the effects of various physical factors. Problem behaviors characteristic of the aged group included 'being soaked in drink' and being inebriated, showing no correlation with the presence or absence of dementia. There was no significant difference in frequency of delirium between the aged group and the younger group. However, in aged alcoholics delirium tended to continue for a longer period during abstinence and was more likely to occur even during non-abstinence. A similar trend was found in aged alcoholics with dementia compared with those without dementia. PMID- 9201758 TI - Effective electroconvulsive therapy for stupor in the high risk patient: a report of two cases. AB - We report two cases of stupor in which the patients were safely treated by electroconvulsive therapy (ECT) despite high risk conditions. Case 1 was a 72 year old schizophrenic woman who had developed catatonic stupor and had joint contractures as a complication of rheumatoid arthritis. Case 2 was a 52 year old woman who developed a stuporous state which was complicated by severe dehydration with hypernatremia. In both cases, psychotic symptoms were improved by ECT without event. Careful application of ECT seemed to be effective and safe even for stupor in high risk patients. PMID- 9201757 TI - Cavum septum pellucidum in schizophrenia: a magnetic resonance imaging study. AB - In order to determine if cavum septum pellucidum (CSP) is more prevalent in schizophrenic patients, we studied 72 Japanese patients who fulfilled the DSM-III R criteria for schizophrenia and 41 normal controls. Sagittal, 1 mm thick magnetic resonance imaging slices of the entire cranium were obtained using a gradient-echo pulse sequence, and coronal and axial images were reconstructed for assessment. A CSP was observed in 34 patients (47.2%) and in 16 controls (38.0%). Although the CSP appeared to be more prevalent in schizophrenic patients, this difference was not statistically significant. However, schizophrenic patients with a history of long-term institutionalization had a higher incidence of CSP compared with patients who had not been admitted to hospital for more than 3 years (68.2 vs 38.0%). These results suggest that the CSP may be a pathophysiology that characterizes schizophrenic patients with poor prognoses. PMID- 9201759 TI - Trazodone for aggression in an adolescent with hydrocephalus. AB - A case of 19 year old male with hydrocephalus is reported whose aggressive self injurious behaviors were resistant to conventional pharmacotherapy but successfully treated by trazodone. In addition to the self-injurious behaviors, this patient displayed withdrawal and eating refusal, which initially resulted in his admission to a psychiatric ward. Various conventional treatments with pharmacotherapy (e.g., tricyclic antidepressants, antipsychotics, anxiolytics and anticonvulsants) in combination with psychotherapy and family therapy proved not to be effective for 15 months. Neither was electroconvulsive therapy successful. Administration of trazodone for 5 months after tapering of the above agents improved his aggressive behaviors. A survey of previous cases with organic brain syndromes who had aggressive behaviors and responded well to trazodone revealed that most of the cases were aged individuals and that cases in adolescence are rare. PMID- 9201760 TI - Immunohistochemical examination of phosphorylated tau in granulovacuolar degeneration granules. AB - Granulovacuolar degeneration (GVD) and neurofibrillary tangles (NFT) are neuropathological features in Alzheimer's disease (AD). The molecular mechanism of GVD formation remains unknown. Recent immunohistochemical investigations suggested a potential link of NFT to GVD formation. Enzyme histochemical studies and electronmicroscopic findings suggested that GVD is formed through lysosomal autophagy of intraneuronal substances. We recently demonstrated that in non demented cases NFT was phosphorylated at serines 199, 202 and 422 in paired helical filament (PHF)-tau more than in serine 396, while NFT in AD cases was similarly phosphorylated at these four sites in tau. In this study, we demonstrated immunohistochemically a similar phosphorylation state of tau in GVD granules to that in NFT in both non-demented cases and AD patients by using a mouse monoclonal anti-tau antibody and three phosphorylation site-specific antibodies for PHF-tau, indicating that GVD granules and NFT are composed of similar phosphorylated-tau. However, we could not detect PHF structures within any GVD using electronmicroscopy, indicating that PHF itself is not phagocytized by lysosomes during GVD formation. Therefore, the source of GVD granules might be phosphorylated pre-PHF-tau. PMID- 9201761 TI - Effect of chronic ipsapirone treatment on the density of 5-HT1A receptors and 5 HT1A receptor mRNA in discrete regions of the rat brain. AB - There is increasing evidence that the 5-hydroxytryptamine (HT)1A partial agonist ipsapirone is an effective anxiolytic/antidepressant agent, although its mechanism of action is not clear. In this study, we investigated the effects of chronic ipsapirone treatment (5 or 10 mg/kg; twice daily, 3 weeks) on 5-HT1A receptor density 8-hydroxy-2-(di-n-propyl amino) tetralin (8O H-DPAT) binding and the level of its mRNA (in situ hybridization) in various regions of the rat (male Wistar 250 g) brain. Receptor density was reduced in the frontal cortex, but did not change significantly in the hippocampus and dorsal raphe nucleus. The level of receptor mRNA was unchanged in each of these brain regions. The present results suggest that the clinical anxiolytic effects of ipsapirone may be mediated partly by postsynaptic action on serotonergic transmission in the frontal cortex, but not in the hippocampus or dorsal raphe nuclei. PMID- 9201763 TI - Transient elevation of nerve growth factor content in the rat hippocampus and frontal cortex by chronic ethanol treatment. AB - The nerve growth factor (NGF) content in the hippocampus and frontal cortex of chronic ethanol-treated rats was measured and compared with that of control rats, using a two-site enzyme immunoassay (EIA) system. The different time periods of chronic ethanol treatment caused transient elevation of the NGF content in both the hippocampus and frontal cortex. The NGF content in the hippocampus was significantly elevated in rats undergoing ethanol treatment of 2 weeks and 1 month. Nerve growth factor content of the 1 month treatment was higher than that of the 2 week treatment. However, a 3 month administration of ethanol reduced the NGF content to the control level. The NGF content in the frontal cortex increased significantly in the 2 week administration, but decreased to the control level in the 1 month administration. The increase of NGF may be caused by the proliferation of glial cells or the enhancement of neuronal production of NGF. PMID- 9201762 TI - Magnesium ion augmentation of inhibitory effects of adenosine on dopamine release in the rat striatum. AB - The effects of adenosine and magnesium ion (Mg2+) on striatal dopamine release were studied in awake rats by in vivo microdialysis. The mean striatal basal levels of dopamine release at Mg2+ free perfusate were 56.95 +/- 5.30 fmol/sample (for 20 min). By varying the Mg2+ levels in perfusate from 0 mmol/L to 1, 10 or 40 mmol/L, the dopamine release was inhibited by Mg2+ in a level-dependent manner. Perfusion with modified Ringer's solution containing zero Mg2+ and from 5 to 50 mumol/L adenosine, non-selective adenosine agonist, as well as 0.1 mumol/L 2-chloro-N6-cyclopentyladenosine (CCPA), selective adenosine A1 agonist, showed no effect on dopamine release. However, from 5 to 50 mumol/L adenosine and from 0.1 to 1 mumol/L CCPA plus Mg2+ (1 and 40 mumol/L) perfusion decreased the dopamine release. This inhibitory effect of adenosine and CCPA on striatal dopamine release was enhanced by an increase in extracellular Mg2+ levels. Levels of 50 mumol/L of 8-cyclopentyl-1,3-dimethylxanthine (CPT), a selective adenosine A1 receptor antagonist, in perfusate increased the dopamine release under conditions both with and without Mg2+. This stimulatory effect of CPT on striatal dopamine release was reduced by an increase in extracellular Mg2+ levels. As a result, CPT antagonized the inhibitory effects of adenosine and CCPA on dopamine release under conditions of the presence and absence of Mg2+. These results suggest that the inhibition of striatal dopamine release by adenosine was mediated by adenosine A1 receptor. This inhibition was intensified by Mg2+. This study also revealed that the concentrations of Mg2+, which ranged from physiological to supraphysiological, reduced the striatal dopamine release; furthermore it was found that the physiological concentration of Mg2+ potentiated the effects of adenosine agonists, but inhibited adenosine antagonist. Thus, the present study, using in vivo microdialysis preparations, suggests Mg2+ inhibits the calcium ion channels and enhances the adenosinergic function in the central nervous system. PMID- 9201764 TI - Increased striatal glutamate transporter by repeated intermittent administration of methamphetamine. AB - We have examined the immunoreactivities of a glutamate (Glu) transporter, GLT-1, in rat brains treated with a single or repeated intermittent administration of methamphetamine (MAP). In the stratum of behaviorally sensitized rate induced by repeated intermittent MAP treatment, GLT-1 immunoreactivities were increased by 51%. There was no difference in the GLT-1 immunoreactivities in all regions examined between rate treated with a single administration of MAP and the control rats. These results suggest that hyperglutamatergic activity in the striatum is involved in the induction of behavioral sensitization caused by MAP. PMID- 9201766 TI - A spectrum for obsession and personality disorders. AB - Obsession was introduced by Kraepelin in 1915 and has been studied extensively since. When a person with obsession becomes physically exhausted with chronic rumination accompanied by suspicion, he or she is driven to impulsive acts, and develops a personality disorder that displays persistent abnormal activities. Obsession is related closely to depression and schizophrenia. Obsession is induced when uncertainty and instability dominates intelligence and creativity. The current social hierarchy of a strongly controlled society rejects diversity of humanity and often triggers personality disorders. This article reviews obsession and a myth as primitive mentality, normal and abnormal obsession, obsession vs possession, society and obsession/ impulsion/degeneration, obsession and slowness/autism, a recent biological approach to obsession and a spectrum for obsession. PMID- 9201765 TI - The concept of 'atypical psychoses': special reference to its development in Japan. AB - Reviewing the development of concepts of 'atypical' psychoses in European countries and in the United States shows that there are various terminologies which are given to a group of psychoses unclassifiable within Kraepelinian dichotomy. Bouffee delirante (French school), cycloid psychoses (Leonhard, Perris), reactive psychoses (Scandinavian school) and acute schizoaffective psychoses (Kasanin) are the most common terms. These are consistent in terms of acute onset, polymorphic symptomatology and good prognosis, and are considered to be distinct from major psychoses, especially from typical schizophrenia. The concept atypical psychoses in Japan was developed under the influence of Mitsuda's clinico-genetic studies. According to Mitsuda, atypical psychoses are not mere phenotypical variants of typical schizophrenia and manic-depressive psychosis (MDP) but belong to a genetically different category and are probably heterogeneous. The characteristic features in the Japanese concept of atypical psychoses emphasizes the alteration of consciousness in symptomatology and pays attention to the nosological relationship with epilepsy, as well as with schizophrenia and MDP. Thus, in Japan it is generally considered that atypical psychoses are independent of 'typical' major psychoses and are located nosologically in the border area between typical schizophrenia, MDP and epilepsy. PMID- 9201767 TI - Relationship between solvent inhalation and antisocial behavior: special emphasis on two types of violence seen in solvent abusers. AB - Organic solvent inhalation is a serious problem among youths in Japan. It induces physical and mental disorders, and is related closely to crime and delinquency. The relationship between solvent inhalation and antisocial behavior was investigated in 75 youths. The subjects were divided into three groups according to the level of violent behavior and the time of appearance: a non-violent group, a late-onset group (violence occurred after the start of inhalation) and an early onset group (violence had occurred before the start of inhalation). Various parameters were compared among the groups. The late-onset group showed the following characteristics; (i) the frequency of inhalation was high, and many subjects experienced hallucinations and mood changes caused by inhalation; (ii) the family environment was characterized by conflict; and (iii) the subjects had strong psychological conflicts and showed dissociative coping with frustration. The violence, psychological conflict and dissociation in this group were found to be related to the inhalation, with familial conflict as a background. The violence in the early-onset group was considered to be the manifestation of a violent personality as the frequency of inhalation and the incidence of mental symptoms were both low. The subjects in this group showed weak psychological conflict and tended to be demanding of others when attempting to cope with frustration. Two types of violent behaviors in the teenagers who inhaled solvents were identified. The violence of solvent inhalers should be managed according to the type. PMID- 9201768 TI - Somatization in adolescence with reference to dysmorphophobia. AB - An adolescent male obtained a position in a large business with assistance from others. Initially he had tension headaches due to maladjustment. He then showed signs of dysmorphophobic symptoms, expressed as a dissatisfaction with his nose. Subsequently, he had cosmetic rhinoplasty twice for esthetic reasons. After a few years, he presented with a psychosis. During treatment his symptoms developed into headache and fever that disappeared upon recovery. He was therefore able to seek employment independently. This transition of symptoms assumes an aspect of adolescent mentality and presents a trial and error approach to establishing a social identity. In general, somatization is assumed to be an immature defense mechanism of individual psychopathology. However, somatization may be a sign of an improvement in one's health. Somatization in adolescence may also be a sign of the beginning of a reintegration into society and have a bridging functional aspect that induces socialization in adulthood. PMID- 9201769 TI - Olfactory evoked potentials in Parkinson's disease, Alzheimer's disease and anosmic patients. AB - Olfactory evoked potential (OEP) recordings were undertaken using amyl acetate stimulation in 20 patients with Parkinson's disease, nine patients with Alzheimer's disease, seven patients with olfactory dysfunction with no other neurological disorder, and 17 control subjects. In order to eliminate the somatosensory factor from the combined somatosensory and olfactory components produced by amyl acetate stimulation, we substracted the potentials using odorless air from those using amyl acetate. In normal subjects, three components were observed, the mean latencies of which were 309 +/- 46, 484 +/- 61 and 710 +/ 55 ms. In all subjects with anosmia (n = 7), no responses were observed. In the patients with Alzheimer's disease, the components were fewer despite having no olfactory dysfunction. In the 20 patients with Parkinson's disease, eight [corrected] patients showed no components, seven patients showed one component and four [corrected] patients showed two components. The components rarely were detected in spite of whether the patients had olfactory dysfunction or not. Olfactory evoked potentials are useful in detecting olfactory dysfunction and the early stages of Alzheimer's disease and Parkinson's disease. PMID- 9201770 TI - Higher cerebral dysfunction in a case with atypical multiple sclerosis with concentric lesions. AB - A patient with atypical multiple sclerosis (MS) with clear concentric structure was studied using high field magnetic resonance imaging (MRI). This case was considered to be Balo's concentric sclerosis. Magnetic resonance imaging showed diffuse multiple concentric demyelinating lesions in the bilateral centrum semiovales, which finally regressed with the necrotic lesions remaining when the patient was discharged. During his clinical course, he showed some higher cerebral dysfunctions, such as memory disturbance, constructual apraxia and acalculia. He was treated with glycerin, prednisone and rehabilitation; all of which were effective in his recovery. Over a 4 month period, the patient recovered clinically, but some intellectual impairment remained. PMID- 9201771 TI - Clinical studies of pervasive developmental disorders in Japan. AB - Articles on pervasive developmental disorders (PDD) published mainly by Japanese child psychiatrists in international journals for the last 20 years were reviewed for the purpose of clarifying the accomplishment and aims of Japanese PDD research. Although Japanese child psychiatrists investigated PDD in various specialties, their contributions to international archives were much fewer than those of Japanese professionals in other branches of medical sciences. This may be accounted for by the absence of an authorized education system of child psychiatrists and strong clinical orientation together with some reluctance of Japanese child psychiatrists to perform research. However, the epidemiology of PDD subtypes, the speech loss in PDD and the psychopathology of persons with high functioning PDD seem to be providing promising research areas for Japanese child psychiatrists based on their clinical experiences. PMID- 9201772 TI - Attitudes of medical students towards persons with mental disorders: a comparative study between Japan and Thailand. AB - This study was conducted at three universities, two in Japan and one in Thailand, in order to elucidate the effects of medical education, especially with regard to contact experience on medical students' attitudes toward persons with mental disorders. Questionnaires, which included the Attitudes Towards Disabled Persons Scale (ATDP) and the Contact with Disabled Persons Scale (CDP), were distributed to 1st year students prior to the commencement of their medicine/psychiatry studies and distributed to 6th (or 5th) year students who had completed their psychiatric curriculum. The ATDP scores were lower for 6th year students at all universities, suggesting that post-education students had a more unfavorable attitude than pre-education students. Thai students indicated more unfavorable attitudes than did the Japanese students. Three factors were extracted from the ATDP scale and termed: negation of character, negation of ability and affirmation of normality. Four factors from the CDP scale were extracted and labeled intimate contact experience, ordinary contact experience, unpleasant contact experience and pleasant contact experience. Greater negative attitudes of post-education students than pre-education students were thought to attribute mainly to an increase in factor score of negation of ability and this result was correlated with an increase in factor score of ordinary contact experience in post-education students. Of the three ATDP factor scores, the higher score of Thai students for negation of character contributed to their overall unfavorable attitude scores. The cross-national similarities and differences of students' attitudes towards and contact experience with mentally disordered persons were discussed from the viewpoint of medical education. PMID- 9201773 TI - Specificity and developmental consequences of speech loss in children with pervasive developmental disorders. AB - Speech loss (SL) was compared in 276 children who had pervasive developmental disorders (PDD) with 62 children with intellectual disabilities without PDD. Speech loss seems relatively specific to PDD because it is significantly more common in children with PDD (26.1%, 72/276) than in those with intellectual disabilities (1.6%, 1/62). In three PDD categories, speech loss occurred in all the 12 children with disintegrative psychosis, 35/149 (23.5%) children with infantile autism and 25/115 (21.7%) children with other PDD. Children with pervasive developmental disorders and speech loss had spoken significantly earlier yet developed less satisfactorily after speech loss than those without it. Speech loss seems fairly specific to PDD and is indicative of unfavorable intellectual development in children with PDD. PMID- 9201774 TI - The clinical features of Tourette's disorder with obsessive-compulsive symptoms. AB - Twenty-three patients with Tourette's disorder (13 with obsessive-compulsive symptoms [OCS] and 10 without) were comparatively investigated. In contrast to OCS-free Tourette's disorder patients, those with OCS were found to be characterized by (i) a higher incidence of volatile temper, (ii) a higher incidence of compulsive tics, (iii) a higher incidence of perinatal disorders and brain wave abnormalities, (iv) a higher severity as rated using the Severity Scale, and (v) a higher prevalence of complications, especially of developmental disorders. Of the subjects with OCS-accompanied Tourette's disorder, approximately half had developed OCS by the onset of tics. These findings suggest the likelihood that OCS-accompanied Tourette's disorder is more strongly associated with organic cerebral disorders, independently of sites of tic disorders, than is OCS-free Tourette's disorder. PMID- 9201775 TI - Erythrocyte deformability in schizophrenic patients. AB - Erythrocyte deformability as a clinical indicator of microcirculatory disturbance was determined in the erythrocyte of 25 schizophrenic patients and of 18 normal controls. Schizophrenic patients had significantly lower erythrocyte deformability than did the normal controls (P < 0.001). This result suggests that microcirculation in schizophrenic patients is disturbed, and that this disturbance might be involved in the pathophysiological genesis of schizophrenia. PMID- 9201776 TI - A multicenter study of sleep-wake rhythm disorders: clinical features of sleep wake rhythm disorders. AB - A multicenter study of sleep-wake rhythm disorders (i.e. non-24 hour sleep-wake syndrome; non-24), delayed sleep phase syndrome (DSPS), irregular sleep-wake pattern (irregular sleepers), and long sleepers, was conducted with the co operation of 25 institutions. One hundred and twenty-one primary sleep-wake rhythm disorders were diagnosed and were classified as 13 non-24, 90 DSPS, 12 irregular and six long sleepers. The mean onset age was about 20 years old and psycho-social factors associating the onset of the disorder were identified in 36% of these patients. The major factors of sleep-wake disorders were personal relationships, advancing to a higher level education, gaining employment, and changes in environment. Most patients were 'night active' prior to appearance of their symptoms. Increase in night activities of modern society seem to result in the occurrence of such sleep-wake rhythm disorders. PMID- 9201777 TI - A multicenter study of sleep-wake rhythm disorders: therapeutic effects of vitamin B12, bright light therapy, chronotherapy and hypnotics. AB - One hundred and six subjects with primary sleep-wake rhythm disorders [13 non-24 hour sleep-wake syndrome (non-24), 76 delayed sleep phase syndrome (DSPS), 11 irregular sleep-wake pattern (irregular) and six long sleepers] were treated with vitamin B12, bright light, chronotherapy and/or hypnotics. These therapies caused moderate or remarkable improvement in 32% of the non-24, 42% of DSPS, 45% of irregular and 67% of long sleepers. A lack of adequate sleep, unpleasant feelings at waking and daytime drowsiness were also improved in DSPS. PMID- 9201778 TI - Diagnostic use of daytime polysomnography versus nocturnal polysomnography in sleep apnea syndrome. AB - The usefulness of daytime polysomnography (DPSG) in the diagnosis of sleep apnea syndrome (SAS) is examined. Diagnostic use was investigated by conducting DPSG of two different time periods (Group M, 11.00-14.00 h, and Group A, 15.00-18.00 h). The subjects were 30 patients (28 men and two women; mean age, 54.0 years). Nocturnal polysomnography (NPSG) and DPSG were investigated by comparing indices of sleep, apnea index (AI) and arterial oxygen saturation (SaO2). There was no significant difference among these indices but there was a significant positive correlation between NPSG and DPSG in all variables related to sleep apnea. Moreover, there was no significant difference in the frequency of each type of apnea between NPSG and DPSG in either group. These findings suggest that DPSG is useful not only in diagnosing SAS but in evaluating its severity. PMID- 9201779 TI - Effects of caffeine on event-related potentials: comparison of oddball with single-tone paradigms. AB - We investigated the acute effects of caffeine (500 mg) on event-related potentials (ERP) in 10 healthy subjects using standard oddball and single-tone paradigms. Event-related potentials were recorded before oral ingestion of caffeine or placebo and 30 min and 210 min after. The oddball paradigm, but not the single-tone paradigm, showed that the P300 amplitude and the area were significantly increased 30 min after ingestion of caffeine and significantly decreased 30 min after ingestion of placebo. The effects of caffeine disappeared at 210 min. Neither the P300 latency nor the reaction time changed significantly with the oddball paradigm. However, the reaction time was shortened 30 min after ingestion of caffeine with the single-tone paradigm. These findings suggest that the caffeine-induced increase in the P300 amplitude may have resulted from the increased allocation of attentional resources to the discriminating process which was not, however, accompanied by facilitation of the process and that caffeine may specifically stimulate the discriminating process involved in the oddball paradigm. In addition, the simple psychomotor performance of buttonpressing in response to a tone signal was accelerated by caffeine. PMID- 9201780 TI - Effects of entorhinal cortex lesion on learning behavior and on hippocampus in the rat. AB - The initial stage of Alzheimer's disease is characterized by a neuropathological change in the entorhinal cortex. In a previous study it was shown that rats with excitotoxic lesion of entorhinal cortex showed an impaired acquisition of passive and active avoidance responses. In this study a rat with excitotoxic lesion of the entorhinal cortex was tested for 'more operant' behavioral learning (i.e., positive reinforcement operant learning). The hippocampus was also examined histologically as acetylcholinesterase-stained sections, and as synaptophysin immunostained sections and examined biochemically by liquid chromatography. Eight weeks after operation, the bilateral entorhinal cortex lesioned rats showed an impaired acquisition of positive reinforcement operant learning. The lesioned side of unilateral entorhinal cortex lesioned rats showed a decrease of acetylcholinesterase-positive fibers in the CA3, the dentate gyrus, and of synaptophysin-positive substances in the CA3. Biochemical study showed a decreased level of acetylcholine in the CA3, and in the dentate gyrus. The histological and biochemical findings are interpreted as indicating that the entorhinal cortex of the rat provides the major extrinsic synaptic input to the hippocampal formation via the circuit which serves as a relay passage through the dentate gyrus and via direct projections into the hippocampus. Behavioral findings confirmed the importance of the entorhinal cortex in memory acquisition and indicated that rats with a partial neuronal loss in the entorhinal cortex may be a useful model for the memory disturbance of Alzheimer's disease. PMID- 9201781 TI - Non-existence of a positive correlation between urinary levels of alpha 1 microglobulin and ulinastatin in patients with Parkinson's disease. AB - Urinary levels of alpha 1-microglobulin (alpha 1M) and of ulinastatin (UT) and the alpha 1M/UT ratio did not differ significantly between age-matched controls and patients with Parkinson's disease, and among subdivided groups based on Yahr's stages in Parkinson's disease. Furthermore, these indexes did not correlate with Yahr's stages. Although alpha 1M and UT levels did not correlate in patients with Parkinson's disease, a positive correlation was observed in the control group. The non-existence of a positive correlation between alpha 1M and UT levels distinguishes Parkinson's disease from other neuropsychiatric diseases such as dementia (Alzheimer-type and vascular dementia), schizophrenia and mood disorder. PMID- 9201782 TI - Reliability of the Japanese version of the Inventory to Diagnose Depression. AB - The reliability of the Japanese version of the Inventory to Diagnose Depression (IDD) which is a self-report to diagnose major depressive disorders (MDD) of DSM III-R, was investigated in 30 cases with MDD and 30 control subjects. On test retest reliability, the agreement of diagnostic performance was substantial (kappa = 0.64, P < 0.001) for 60 subjects, and scores of total and individual items correlated significantly (P < 0.001) between test and retest except for decreased energy, decreased interest, and decreased concentration. The average score of the total IDD severity at test (38.4) was significantly higher than that at retest (28.0; P < 0.01). However, excluding the recovered 10 cases, there was no significant difference seen on the average total score between test and retest (38.8, 30.1, respectively). Internal consistency (Cronbach's alpha = 0.80) and split-half reliability (0.79) were sufficient, and item-total correlations of the IDD were significant (P < 0.01) except for weight gain. The IDD might be useful as a screening tool and for clinical evaluation of subjects in Japan; however, it is necessary to examine the validity of this instrument. PMID- 9201783 TI - Longitudinal change in youth suicide mortality in Okinawa after World War II: a comparative study with mainland Japan. AB - Okinawa prefecture has a unique socio-cultural status in Japan including the experience of having been occupied by the USA from the end of World War II to 1972. In this study, the longitudinal change in youth suicide mortality for those aged 10-29 years in Okinawa (1960-90) was compared with that for the same sex-age groups in mainland Japan (1950-90). In contrast with mainland Japan, no dramatic change in the youth suicide mortality was observed in Okinawa in the 1960s. The rise and fall of teenage suicide mortality in Okinawa during the 1970-80s might be associated with 'reversion anxiety', rather than with the traumatic experience of World War II itself. This seems to be inconsistent with previous speculation regarding the change in youth suicide mortality in mainland Japan. The suicide mortality for men aged 20-29 in Okinawa was significantly higher than that for the same sex-age group in mainland Japan through the observed period. The possible effects of the USA occupation, economic anomie or migration on the suicide in Okinawa should be further examined. PMID- 9201784 TI - Age differences of psychiatric inpatients presenting with physical complications. AB - We reviewed the records of 292 inpatients in the psychiatric ward of Kagoshima University Hospital who were referred from other medical facilities over a 5-year period in order to clarify age differences in the reason for referral. Patients were classified into groups of physically and mentally ill individuals based on indications for admission. Both groups were further divided into four subgroups based on age. The incidence of inpatients with physical illnesses increased with age. Conditions related to pregnancy, childbirth and the puerperium occurred at high frequency in female patients in the 20- to 39-year-old subgroup. Individuals in the 40- to 59-year-old and in the > or = 60 years subgroups suffered more frequently from neoplasms. The proportion of patients manifesting a defective state in all age subgroups with the exception of the under 19-year-old subgroup was significantly higher in the physical illness group than in the mental illness group. The proportion of patients in a depressive state in the > or = 60 years subgroup was significantly higher in the mental illness group than in the physical illness group. Hence, it is necessary to find a method to be able to cope with psychiatric patients with physical complications to solve this problem. PMID- 9201785 TI - Survey of the living conditions and psychosomatic states of the elderly in Japan who receive home help. AB - The living conditions and psychosomatic states of 2828 elderly people receiving care from home helpers were investigated. The physical condition of men was significantly lower than that of women. With regard to daily life (i.e. eating, sitting, standing, excretion, dressing, bathing, walking, tidiness) 70% were rated as being self-supporting. Eighty per cent of subjects were judged as having normal intelligence. Twenty-two per cent of subjects had more than one psychiatric symptom (e.g. memory impairment, insomnia, talking to oneself, and delusions). Among the various medico-welfare supports, home help was recommended most often. PMID- 9201786 TI - Periodicity and prediction of mania onsets in biopolar I affective disorders. AB - In studying the periodicity of mania onsets, cycle-oriented diagrams were made of the clinical course from 257 manic episodes analyzed retrospectively in 34 bipolar I manic-depressive patients for a period of about 5 years. Using these diagrams, the frequent period of mania onsets located in one-quarter of the follow-up period was pre-estimated, and the accordance ratio during a 25 month follow-up period was analyzed. The accordance ratio in all subjects was 39% (11/28) for the first episode and 35% (7/20) for the second episode. These ratios were not significantly different from the expected level (25%). The number of subjects was limited to 11 patients (10 rapid and 1 non-rapid cyclers) whose number of episodes used for the determination of the index cycle was eight or more. The accordance ratio was 64% (7/11) and 60% (6/10) for the first and second episodes, respectively. Their levels were significantly higher than those expected. Periodicity of mania onsets existed at least in rapid cyclers with abundant past data. PMID- 9201787 TI - Functional psychosis mimicking acute confusional state: longitudinal neuropsychological assessment of an acute and transient psychotic patient. AB - One patient with acute and transient functional psychosis was assessed repeatedly using a brief neuropsychological assessment during his recovery from the psychotic episode. The psychotic features of the patient were characterized by perplexed behavior, attentional disturbance and emotional turmoil. Characteristic findings, including impairment of attention tests, dysgraphia and constructional disturbances, were seen. Findings improved in accordance with recovery on a behavioral level. We discussed the similarity of neuropsychological and behavioral abnormalities of this patient and those of patients in an acute confusional state. PMID- 9201788 TI - Brief reactive psychosis induced by sensitivity training: similarities between sensitivity training and brainwashing situations. AB - Sensitivity training (ST), which originated in the USA during the late 1940s, has been used as part of training seminars in Japanese corporations since the late 1950s. The possibility of negative psychiatric effects of ST, and especially its role in inducing psychiatric symptoms, is yet to be clarified. A case of a 41 year-old male company worker whose brief psychosis was induced by a sensitivity training seminar held by the company he worked for is presented. In reviewing the psychopathology of the case with records from the ST seminar, we found similarities between the patient's ST seminar and brainwashing situations. Specifically, the patient experienced severe conflict (of thought process) between his Christian beliefs and being labeled a coward at the seminar. We conclude that monitoring of the ST programs is crucial in order to ensure the psychological safety of ST participants in Japan. PMID- 9201789 TI - The wish to be held: female patients with borderline personality disorder. AB - Two female patients expressed in psychotherapy their wish to be held. This reflects their pathology, intense loneliness and defective sense of self and reality. The 'holding' function of the psychotherapist can place the wish to be held into the mutual interaction even if it seems unacceptable, similar to the holding in mother-child communication. The deliberate efforts by the psychotherapist to 'hold' lead to an improvement of the pathology, and contribute to form an endurable therapeutic relationship. Unavoidable body contact with patients who are in regressive states is discussed, and the need to re-establish the communicative value of contact experiences is emphasized. PMID- 9201790 TI - Relationship between odor perception and depression in the Japanese elderly. AB - Odor perception has been studied in patients with various mental disorders; however, no consensus has been reached as to its detection, identification, or pleasantness/unpleasantness of odors especially in patients with depression. One hundred and nineteen normal elderly individuals living at home were exposed to odors of rose, perfume, white ginger, Indian ink, cigarette smoke, milk, feces and orange scent using the scratch and sniff method. They were asked to rate the strength of each odor, its pleasantness or unpleasantness, their liking for it, and their familiarity with it. They were also asked to complete a self-rating depression scale (SDS). The relationship of the score of each psychological olfactory scale with the SDS score and the difference in the score of each psychological scale between high-SDS and low-SDS groups are discussed. PMID- 9201791 TI - The differences of self-ratings of sleep quality associated with epinephrine and wake time during 4 hour sleep. AB - The present study examined the differences of self-ratings of 4 h sleep in three states: L-WE, where the percentage of waking time and urinary epinephrine are low (< 20% waking time); H-W, where the percentage of waking time and epinephrine levels increase along the basal regression line as determined by a previous study (20-100% waking time and < 7 ng/min); H-E, where epinephrine levels increase more than expected from the basal regression line for the two parameters (> 7 ng/min). Eight healthy male subjects participated twice in a 4 h polysomnograph experiment with four types of sleep onset (total of 64 observations). In group L-WE (52 observations for eight subjects), there were no excessively negative feelings on sleep latency, sleep depth, and feelings of sleep compared with usual sleep according to the questionnaire. Subjective sleep diagrams in group L-WE were similar to polysomnographic findings. Thus, group L-WE was thought objectively and subjectively to have a good sleep state. Groups H-W (eight observations for four subjects) and H-E (four observations for two subjects) had negative feelings regarding sleep depth and feelings of sleep compared with usual sleep. Approximately half the group H-W underrated their sleep compared with objective diagrams, while all cases in group H-E remarkably underrated their sleep in the subjective diagrams. The state of remarkable adrenal medullary secretory activity seen in group H-E and that of the slightly increased activity shown in group H-W were included in poor sleep states objectively and subjectively. PMID- 9201792 TI - Ultra-low-field magnetic resonance imaging in upper airways obstruction in sleep apnea syndrome. AB - The hypothesis that the sites of upper airways obstruction (UAO) are varied in a patient with obstructive sleep apnea syndrome (OSAS) among different sleep stages is studied. Four patients with OSAS underwent ultra-low-field magnetic resonance imaging (MRI) with a field strength of 0.064 Tesla provided real-time images and generated less noise and necessitated less strict magnetic isolation compared with conventional high-field MRI. After the fixed end-apneic sleep stage was determined, the polysomnogram was switched off and ultra-low-field MRI was commenced. The effects of continuous positive airway pressure (CPAP) on the upper airway patency in the deepest sleep stage obtained for each patient was assessed. Upper airway obstruction was found at the level of the palatopharynx (PP) at sleep onset extended to the glossopharynx (GP) during rapid eye movement (REM) sleep in two cases and during NREM sleep in one case. This combined PP and GP obstruction was observed from sleep onset and remained unchanged in one case. The patent upper airways were observed during treatment with CPAP during REM sleep in two patients and during stage two of NREM sleep in the other two patients. It can be concluded that the sites of UAO vary in a patient with OSAS in different sleep stages. The results also suggest the use of the ultra-low-field MRI in order to visualize the dynamic and real-time behaviors of the upper airways during sleep in patients with OSAS. PMID- 9201793 TI - Improvement of schizophrenic symptoms and changes in plasma HVA concentrations, plasma anti-D2 and anti-5-HT2 receptor activities with clozapine. AB - In order to investigate the biological mechanisms underlying the clinical efficacy of clozapine, 200 mg/day of clozapine was added to the drug regimens of 19 patients with chronic, anti-psychotic-resistant schizophrenia, and the plasma homovanillic acid (HVA), clozapine concentrations, anti-dopamine D2 and anti serotonin 5-HT2 receptor activities were measured. After 28 days, six patients showed an improvement of more than 20% over baseline Brief Psychiatric Rating Scale (BPRS) scores. Mean plasma HVA concentrations and anti-D2 receptor activities did not change significantly in the entire group or in the six patients showing improvement. However, anti-5-HT2 receptor activities increased significantly in all 19 patients. Changes in BPRS scores did not correlate significantly with changes in plasma HVA or with changes in clozapine concentrations, or with anti-D2 and anti-5-HT2 receptor activities. PMID- 9201794 TI - The influence of physical restraint or fasting on plaque-forming cell response in mice. AB - This study was designed to investigate the effects of 1- and 3-day (16 h/day) physically restrained or fasting on immunological and endocrine responses in CBF1 mice. The influence of stressors on these responses was evaluated using anti sheep red blood cell plaque-forming assay, and by examining T cell subsets, thymus weight and endocrine hormone levels. The results revealed that a significant elevation of the plaque-forming cells (PFC) was found in spleen cells in 1-day restrained mice, that the PFC were conversely suppressed following 3-day physically restrained stress, and that the PFC were not affected by 1- or 3-day fasting stress. Serum levels of norepinephrine were found to be significantly increased only in 1-day physically restrained mice. No change of T cell subsets and thymus weight was found in 1-day physically restrained mice. A significant increase in serum corticosterone levels was elicited in both 1- and 3-day physically restrained mice, and 3-day fasting mice, while increased Lyt2-positive T cell and thymic atrophy were found only in 3-day physically restrained mice. These findings suggest that immune function was differentially affected by the duration and types of stressors. PMID- 9201795 TI - Age-dependent induction and maintenance of sensitization to methamphetamine induced hyperactivity in mice. AB - Repeated administrations of methamphetamine (2 mg/kg, s.c.), 10 times at 3-day intervals, induced ambulatory sensitization in all groups of mice that were 13-, 15-, 19-, 23- and 36-week-old at the start of methamphetamine administration. The most prominent sensitization was observed in the 19-week-old mice. Among five groups of mice, even though the mice of 36 weeks old showed the highest sensitivity to methamphetamine at the first administration, they exhibited the lowest sensitization during the latter stage of repeated methamphetamine administration. Methamphetamine sensitization once established was well reproduced by the post-sensitization period of 8 weeks. Furthermore, the group of mice given methamphetamine with post-sensitization interval of 8 weeks (19-week old mice) exhibited further enhancement of the sensitization. In contrast, the groups of mice given methamphetamine with post-sensitization intervals of 12 and 25 weeks (the 23- and 36-week-old, respectively) showed a significant reduced sensitization, and the latter group failed to reach the level of sensitization previously established. These results suggest that the induction of and maintenance of methamphetamine sensitization are dependent on the age of the mice, and that methamphetamine sensitization once established completely persists for up to 8 weeks. PMID- 9201796 TI - Psychomotor effects of the anxiolytic abecarnil: a comparison with lorazepam. AB - Abecarnil is a metabolically stable beta-carboline that binds with high affinity and selectivity to central benzodiazepine receptors. The effects on cognitive and psychomotor skills of abecarnil (ZK 112-119), 2.5 mg and 5.0 mg, were compared with lorazepam 2.0 mg and placebo. Twenty-four healthy, young males participated in a double-blind, four-way Latin square design and performed batteries of behavioral tests at predrug and at 20, 40, 60, 80, 100, 120, 180, 240, 360 and 480 min after drug administration. Abecarnil 5.0 mg and lorazepam 2.0 mg displayed similar impairment profiles in tests of cognitive functions including memory encoding. Abecarnil 2.5 mg was substantially less impairing than lorazepam. Impairment levels of the abecarnil and lorazepam treatments peaked at 2-3 h after oral administration. The two abecarnil doses showed dose-dependent effects on the cognitive and psychomotor tasks. All three drug treatments were well tolerated by the subjects, with no one terminating early due to adverse events. The incidence of reported adverse events for abecarnil was dose dependent. The most frequent, statistically significant adverse effects were drowsiness, lack of concentration and visual disturbance for abecarnil 5.0 mg; and lack of concentration and dizziness for lorazepam 2.0 mg. There were no significant differences in adverse incidence rates between abecarnil 2.5 mg and placebo. PMID- 9201797 TI - The attenuating effect of carteolol hydrochloride, a beta-adrenoceptor antagonist, on neuroleptic-induced catalepsy in rats. AB - It is known that beta-adrenoceptor antagonists are effective in the treatment of akathisia, one of the extrapyramidal side effects that occur during neuroleptic treatment. Neuroleptic-induced catalepsy, a model of neuroleptic-induced extrapyramidal side effects, was considered suitable as a model for predicting neuroleptic-induced akathisia in humans, although neuroleptic-induced catalepsy was not considered a specific test for neuroleptic-induced akathisia. Therefore, the effects of carteolol, a beta-adrenoceptor antagonist, on haloperidol-induced catalepsy in rats were behaviorally studied and compared with those of propranolol and biperiden, a muscarinic receptor antagonist. Carteolol, as well as propranolol and biperiden, inhibited the haloperidol-induced catalepsy. The inhibitory effect of carteolol was almost comparable to that of propranolol, but was weaker than that of biperiden. Carteolol did not evoke postsynaptic dopamine receptor-stimulating behavioral signs such as stereotypy and hyperlocomotion in rats. Carteolol did not antagonize the inhibitory effects of haloperidol on apomorphine-induced stereotypy and locomotor activity in rats. In addition, carteolol did not evoke 5-HT1A receptor-stimulating behavioral signs such as flat body posture and forepaw treading and did not inhibit 5-hydroxytryptophan-induced head twitch in rats. Finally, carteolol did not inhibit physostigmine-induced lethality in rats. These results strongly suggest that carteolol improves haloperidol-induced catalepsy via its beta-adrenoceptor antagonistic activity and is expected to be effective in the treatment of akathisia without attenuating neuroleptic-induced antipsychotic effects due to its postsynaptic dopamine receptor antagonistic activity. PMID- 9201798 TI - Morphine-induced long-term sensitization to the locomotor effects of morphine and amphetamine depends on the temporal pattern of the pretreatment regimen. AB - The development of behavioural sensitization is thought to depend on the dose and temporal pattern of drug treatment. Previous studies have shown that two distinct morphine pretreatment regimens cause different long-term neuroadaptations in rat striatum. Therefore, in the present study the ability of these pretreatment regimens to induce long-term behavioural sensitization was investigated. One pretreatment regimen, termed "chronic", consisted of three daily injections, for 5 days, with escalating doses (10-50 mg/kg) of morphine, and the other, termed "intermittent", of 14 daily injections with morphine (10 mg/kg). Both intermittent and chronic morphine pretreatment caused sensitization to the locomotor effects of morphine, 3 weeks post-treatment, although the former induced a far greater level of sensitization. Moreover, 3 weeks post-treatment, intermittent, but not chronic, morphine pretreatment induced cross-sensitization to the locomotor stimulant effects of amphetamine. Behavioural sensitization following intermittent morphine pretreatment was clear-cut both 1 day and 3 weeks post-treatment, while after 9 weeks, the locomotor effects of morphine were still slightly augmented. It is concluded that intermittent morphine pretreatment is far more effective in inducing long-term behavioural sensitization than chronic morphine pretreatment. PMID- 9201799 TI - A double-blind comparison of abecarnil and diazepam in the treatment of uncomplicated alcohol withdrawal. AB - Treatment of the alcohol withdrawal syndrome is best accomplished using pharmacologic agents that have minimal interaction with alcohol, have limited adverse effects, and are without abuse potential. The partial benzodiazepine receptor agonist beta-carboline compound, abecarnil, has been shown in animal and human studies to possess a number of these characteristics and to be useful in the reduction of alcohol withdrawal convulsions in mice. In this study, 49 alcohol-dependent inpatients who exhibited at least moderate symptoms of uncomplicated alcohol withdrawal were treated over a 5-day detoxification period with abecarnil or diazepam and rated daily for alcohol withdrawal symptoms and adverse events. Both the abecarnil and diazepam treatment groups exhibited a similar marked reduction in withdrawal symptoms over time. In addition, similar rates of successful treatment and improvement were observed after 1 day of treatment and at termination in alcoholics treated with either medication. Overall, rates of adverse events and changes in liver enzymes were similar in both treatment groups and were generally benign. Because of the unique pharmacologic profile of abecarnil in animal and in non-clinical human studies, including anticonvulsant action, low abuse liability, and a favorable side effect profile, further study of compounds of the partial benzodiazepine receptor agonist type in the treatment of alcohol withdrawal syndromes seems warranted. PMID- 9201800 TI - Mecamylamine- or scopolamine-induced learning impairment: ameliorated by nefiracetam. AB - Nefiracetam is undergoing preclinical and clinical tests as a cognition-enhancing drug in Alzheimer's disease (AD). Nicotinic cholinergic receptors are lost in AD, and nicotinic as well as muscarinic cholinergic receptors are involved in the modulation of eyeblink conditioning. Experiments were carried out using young rabbits to examine the effect of nefiracetam on cholinergic antagonists to nicotinic (mecamylamine) and muscarinic (scopolamine) receptors. Rabbits were tested for 15 days in the 750 ms delay eyeblink classical conditioning paradigm in paired and explicitly unpaired conditions. Nefiracetam at a dose of 15 mg/kg significantly ameliorated the effects of 0.5 mg/kg mecamylamine, and nefiracetam at a dose of 10 mg/kg significantly ameliorated the effect of 1.5 mg/kg scopolamine. The vehicle alone and nefiracetam alone groups performed similarly to the groups treated with mecamylamine or scopolamine and nefiracetam. Reversal by nefiracetam of a nicotinic as well as a muscarinic cholinergic antagonist indicates that the drug may affect deficits specific to AD. PMID- 9201801 TI - Opioid modulation of attention-related responses: delta-receptors modulate habituation and conditioned bradycardia. AB - Endogenous opioids modulate attention-related heart rate responses evoked by novel stimuli and conditioned signals in ways that differ from their better-known effects on motivation and memory functions. We investigated the role of delta opioids in modulating bradycardiac orienting and Pavlovian conditioned responses in rabbits, following i.v. treatment with the highly selective delta-receptor antagonist naltrindole (NTI; 0.037-0.370 mg/kg). When administered immediately before testing, NTI induced modest but detectable effects: the lowest dose increased cardiac discrimination near the end of the first training session, whereas the higher doses of NTI impaired discrimination, compared to saline treated controls. NTI treatment immediately before testing also appeared to promote habituation of bradycardiac orienting responses elicited by novel tones, but NTI did not alter unconditioned heart rate responses following tone-shock pairs or extinction of conditioned responses. In contrast, the low dose of NTI administered 20 min, rather than immediately before testing, facilitated conditioned bradycardia during extinction, as well as during training. These results provide evidence that endogenous delta-opioid modulators normally delay the disappearance of bradycardiac orienting responses during habituation, inhibit or promote the development of bradycardiac conditioned responses during Pavlovian training depending on dose, and promote the disappearance of conditioned responses during extinction. These findings suggest that endogenous delta-opioid activity, probably involving both peripheral and central systems, coincides with, and may reflect, uncertainty about stimulus significance. PMID- 9201802 TI - The relationship between D2 receptor occupancy and plasma levels on low dose oral haloperidol: a PET study. AB - The purpose of this study was to determine the relationship between dopamine D2 receptor occupancy and plasma haloperidol. Twelve patients treated with 1-5 mg/day of haloperidol had their D2 occupancy measured using [11C]-raclopride and positron emission tomography and haloperidol plasma levels measured using gas chromatograph mass spectrophotometer. The patients exhibited haloperidol plasma levels ranging from 0.5 to 5.8 ng/ml and D2 occupancy from 53 to 88%. The D2 occupancy was related to the plasma level as a saturating rectangular hyperbola relationship (r2 = 0.84) and it showed that, on average, 50% D2 occupancy was achieved with 0.51 ng/ml and 80% D2 occupancy with 2.0 ng/ml. Our findings demonstrate that 2-5 mg/day of haloperidol, which usually leads to plasma levels of 1-2 ng/ml, would be expected to induce 60-80% dopamine D2 receptor occupancy. If, as has been claimed, 70% D2 occupancy is adequate for typical neuroleptic response, then the conventional use of > 10 mg/day may have been too high, since 70% occupancy can be achieved in most patients by 2-5 mg/day. On the other hand, if as others have suggested, 8-12 ng/ml of haloperidol is the correct therapeutic window for plasma levels, then the required therapeutic D2 occupancy is closer to 90%, not 70%. The implications of the D2 occupancy findings for the optimal dosing of neuroleptics are discussed. PMID- 9201803 TI - Effects of flumazenil in the treatment of benzodiazepine withdrawal--a double blind pilot study. AB - Flumazenil, a partial benzodiazepine agonist with low intrinsic activity, was tested for potential use in patients experiencing withdrawal symptoms after traditional treatment for benzodiazepine dependency. On two occasions, separated by 1-13 weeks, ten patients treated for benzodiazepine dependency and ten controls received cumulative doses of flumazenil (0.05, 0.10, 0.25, 0.50 and 1.00 mg at 15-min intervals) or placebo, with assessments of withdrawal symptoms and physiological variables after each dose. As expected, there was an overall difference between patients and controls, with patients scoring higher on negative and somatic items and lower on positive psychological items. Flumazenil reduced symptoms thought to be important in withdrawal in patients treated for benzodiazepine dependency. In contrast to the patient group, controls reacted in the opposite direction with increases in negative experience when given flumazenil. Further research may develop flumazenil as a therapeutic option in the treatment of benzodiazepine withdrawal. PMID- 9201804 TI - Disruption of trace conditioning of the nictitating membrane response in rabbits by central cholinergic blockade. AB - Central muscarinic cholinergic involvement in classical conditioning of eyeblink responses was determined in trace and delay paradigms. Rabbits were trained on a trace procedure in which a 250-ms tone conditioned stimulus (CS) and a 100-ms air puff unconditioned stimulus (UCS) were presented with a 500-ms trace interval. Each training session day consisted of ten tone alone, ten air-puff alone and 80 paired CS-UCS trials. Scopolamine hydrochloride at doses of 0.03 and 0.1 mg/0.5 ml per kg, s.c. dose-dependently disrupted acquisition of conditioned responses. Rabbits that were treated with scopolamine and failed to learn showed a gradual increase in conditioned responses during an additional training period with saline injections and no transfer from earlier training. Scopolamine methyl bromide, which does not appreciably cross the blood-brain barrier, showed no effects in the trace conditioning paradigm at a dose of 0.1 mg/kg, s.c., indicating central cholinergic blockade is responsible for the suppressive effect of scopolamine. Scopolamine hydrochloride at a dose of 0.1 mg/kg, s.c. did not block acquisition in the delay procedure with a 250-ms inter-stimulus interval, although the rate of acquisition was somewhat reduced by the drug. These data are the first to demonstrate that classical conditioning of the eyeblink response in the trace procedure is highly sensitive to central cholinergic deficits. PMID- 9201805 TI - Nitric oxide synthase inhibition blocks phencyclidine-induced behavioural effects on prepulse inhibition and locomotor activity in the rat. AB - The ability of the nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME), to block the behavioural effects of the potent psychotomimetic, phencyclidine, was tested in rats using two different behavioural models. L-NAME was found to block both phencyclidine-induced disruption of prepulse inhibition of acoustic startle and phencyclidine-induced stimulation of locomotor activity. A selective action of L-NAME on the effects of phencyclidine was indicated, since L-NAME did not alter the effects of amphetamine, another potent psychotomimetic, in these behavioural models. These observations suggest that a nitric oxide dependent mechanism may be involved in the effects of phencyclidine in the central nervous system. PMID- 9201806 TI - Non-functional CYP2D6 alleles and risk for neuroleptic-induced movement disorders in schizophrenic patients. AB - The use of classic anti-psychotic drugs in the long-term treatment of schizophrenia is associated with risk for extrapyramidal side-effects, such as akathisia, parkinsonism and tardive dyskinesia (TD). Approximately 5-10% of European Caucasians lack the cytochrome P450 enzyme CYP2D6 (so-called poor metabolizers; PM), which normally metabolizes several drugs including many neuroleptics. PM subjects may achieve high or toxic plasma levels upon standard drug therapy. In this study we have examined 100 subjects from the Nithsdale cohort of schizophrenic patients in South-west Scotland receiving long-term neuroleptic medication, which enabled us to perform both a cross-sectional and longitudinal evaluation of extrapyramidal side-effects in relation to the genetically impaired CYP2D6 metabolism. We identified ten (10%) schizophrenic subjects with the PM genotype. In the cross-sectional study, the prevalence of TD, parkinsonism and akathisia was 51%, 38% and 15%, respectively. Patients with TD or parkinsonism were significantly older than patients without these side effects. In contrast, patients with akathisia were significantly younger than patients without akathisia. There was a non-significant tendency for PM subjects to have more severe ratings for TD and parkinsonism. In the long-term evaluation based on repeated ratings since 1981, there was a non-significant 3-fold higher frequency of PM subjects among schizophrenic patients with longitudinal TD, as compared with the group of patients with fluctuating or no TD. These results indicate that genetically impaired CYP2D6 metabolism may be a contributing factor for the development of persistent TD. PMID- 9201807 TI - Behavioural profiles of the reversible monoamine-oxidase-A inhibitors befloxatone and moclobemide in an experimental model for screening anxiolytic and anti-panic drugs. AB - The present study compared the behavioural effects of acute and chronic (one daily i.p. injection for 14 days) treatments with the reversible monoamine oxidase-A inhibitors (RIMAs) moclobemide (3 and 10 mg/kg) and befloxatone (0.3 and 1 mg/kg) in the Mouse Defence Test Battery (MDTB) which has been designed for screening anxiolytic and anti-panic drugs. In the MDTB, Swiss mice were confronted with a natural threat (a rat) and situations associated with this threat. Primary measures taken before, during and after rat confrontation were escape attempts, flight, risk assessment (RA) and defensive threat and attack. After acute administration of both compounds, no modification of defensive behaviours were observed. This was in contrast to chronic treatments, where moclobemide (3 and 10 mg/kg) and befloxatone (1 mg/kg) produced a significant reduction in one flight measure (avoidance distance when the rat was approaching). In addition, befloxatone (0.3 and 1 mg/kg), but not moclobemide, increased RA responses when mice were constrained in one part of the apparatus facing the rat, which remained at a constant distance. No other drug effects were observed with either compound. Although these behavioural profiles are consistent with an anxiolytic-like effect, the finding of an action upon a limited number of defence responses suggests a weaker anxiolytic-like potential compared to that of classical anxiolytics. However, in view of previous data with panic-modulating compounds on flight behaviours in the MDTB, the present results are in line with clinical results showing that moclobemide is effective in panic disorders and suggest that befloxatone may have some efficacy in the clinical management of panic. PMID- 9201808 TI - Amygdala and hippocampus control dissociable aspects of drug-associated conditioned rewards. AB - Limbic innervation of the nucleus accumbens via the ventral subiculum/hippocampus and basolateral area of the amygdala has been shown to determine dissociable aspects of behaviour controlled by stimuli associated with natural rewards. However, the respective contributions of the ventral subiculum and amygdala to behaviour governed by drug-associated stimuli remain to be determined. Experiments consisted of two phases: drug-stimulus training, and subsequent stimulus-only testing. Initial training sessions were of two alternating forms. During drug sessions, responding upon one lever resulted in an infusion of 1 microgram d-amphetamine into the nucleus accumbens, whilst during saline sessions d-amphetamine was replaced with saline. Each infusion (drug or saline) was preceded with either a light, or tone. Responding upon a control lever had no programmed consequences. Following training, the levers were retracted, and instead two novel vertical bars were extended from the chamber ceiling. Movement of one bar produced the drug-associated stimulus, whilst the alternative bar produced the saline-associated stimulus. Infusions of the AMPA receptor antagonist CNQX into the ventral subiculum or basolateral area of the amygdala (0, 0.2, 2.0 nmol) were made immediately before the start of each session. Intra basolateral area of the amygdala CNQX reduced responding upon the drug-associated stimulus bar, but at the same time increased responding upon the saline associated stimulus bar. By contrast, intra-ventral subiculum CNQX reduced drug associated stimulus responding selectively. Neither manipulation affected levels of activity within the operant chamber extraneous to the bar-pushing response. Hence, the basolateral area of the amygdala appeared to have determined the degree of discriminative control exerted over behaviour by the drug-associated stimulus, whilst the ventral subiculum is suggested to have determined the efficacy of the conditioned reward. PMID- 9201809 TI - Effects of methylphenidate on spatial working memory and planning in healthy young adults. AB - Previous studies of the effects of the psychomotor stimulant, methylphenidate, have concentrated on vigilance and reaction time tasks. In this study, the effects of methylphenidate on more complex aspects of cognition were studied using tasks from the CANTAB battery and related tests which have been shown to be sensitive to frontal lobe dysfunction. Twenty-eight young healthy men participated in a counterbalanced, double-blind, placebo-controlled study of the effects of methylphenidate. Cognitive assessment included tests of spatial working memory, planning, verbal fluency, attentional set-shifting and sustained attention. Methylphenidate had significant effects on performance of the tests of spatial working memory and planning but not on the attentional and fluency tests. When the drug was taken on the first test session, performance on the spatial tests was enhanced by the drug compared to placebo. However, when the drug was taken second, performance accuracy was impaired whereas response latencies were decreased. These results are consistent with a hypothesis that methylphenidate influences performance in two conflicting ways; enhancing executive aspects of spatial function on novel tasks but impairing previously established performance. This pattern of effects is discussed within the framework of dual, interacting arousal mechanisms. PMID- 9201810 TI - A human B-lymphoblastoid cell line constitutively producing Epstein-Barr herpesvirus and JHK retrovirus. AB - The human B-lymphoblastoid cell line, designated JHK-3, with pre-B-cell characteristics, chronically produces two viruses, Epstein-Barr virus (EBV) and JHK virus, an apparently novel retrovirus. The JHK-3 cells are much more productive of extracellular EBV than the high-producer marmoset line B95-8. The extracellular virus of the JHK-3 EBV strain is relatively fragile, more broadly dispersed in an ultracentrifuged sucrose gradient than the B95-8 EBV and more susceptible to disruption by combined treatment with urea and dithiothreitol. By restriction fragment length polymorphism analysis, the JHK-3 EBV strain resembles the EBV strain FF-41. The JHK-3 cells also produce an incompletely characterized, relatively fragile, enveloped, icosahedral RNA virus that contains Mn(++) dependent reverse transcriptase. JHK virions measure 85 nm in ultrathin sections, much smaller than other Retroviridae. The JHK virus exhibits a distinctive morphogenesis, most nearly resembling C-type retroviruses. The JHK-3 cell line provides a human cell model for investigating virus/virus interactions and their pathogenetic affects on host cells which chronically and simultaneously produce DNA and RNA viruses. PMID- 9201811 TI - Inhibition of prokaryotic cell growth by HIV1 Vpr. AB - We have cloned the nef, vif, vpr and vpu genes of HIV1 in the pGEX system to produce auxiliary proteins of HIV1 as N-terminal fusions with glutathione S transferase (GST). Some GST proteins are difficult to obtain under standard conditions. The synthesis and solubility varied considerably from one protein to another. We investigated the reasons for the poor production of GST-Vpr, GST-Vpu and GST-Vif. Interestingly, using this GST prokaryotic model, we demonstrated that Vpr, which is known to block the cell cycle of mammalian and yeast cells at the G2 phase, is also bacteriostatic for Escherichia coli. The effect on E. coli was specific to Vpr, and was not linked to the expression of the other HIV1 proteins. This suggests that Vpr interferes with components of cell replication that are conserved from prokaryotes to eukaryotes. Thus, E. coli appears to be a convenient model system for studies on the function of Vpr. PMID- 9201812 TI - Evaluation of HIV1 infection status by HIV1 PCR and culture methodologies in a small cohort of intravenous drug users. AB - A small cohort of high-risk intravenous drug users (IVDU) from the Baltimore, MD, area was evaluated for HIV1 infection status and viral load. Quantitative dilution endpoint HIV1 DNA polymerase chain reaction (PCR) results, from HIV proviral DNA from quantitated peripheral blood mononuclear cell (PBMC) lysates, were compared to the dilution endpoint results for HIV PBMC micrococulture. The quantitative dilution endpoint HIV1 PCR was more rapid, sensitive and reproducible. In addition, an HIV1 capture RT-PCR technique was used to qualitatively detect the presence or absence of intact HIV1 virus in IVDU plasma and was compared with plasma culture detection, for HIV1 viraemia. Using the results of the PCR techniques, a rapid molecular assessment of the HIV1 infection status can be attained, which is important, as the IVDU population can be difficult to study prospectively. The PCR techniques can also be used to assess HIV1 burden as well as the potential effectiveness of antiviral therapies. These molecular techniques can be used to monitor the progression of HIV in patients and to evaluate the clinical effects of concurrent substance abuse. PMID- 9201814 TI - Genotyping of hepatitis C virus performed by type-specific PCR in comparison to nucleotide sequencing of NS5 and core regions. PMID- 9201813 TI - PCR-based detection and typing of human adenoviruses in clinical samples. AB - Since human adenoviruses (Ad) are associated with a variety of diseases, there is need for a fast and sensitive diagnostic procedure. The polymerase chain reaction (PCR) has been previously applied for the detection and typing of adenoviruses directly in clinical samples. So far, only Ad8, Ad31, Ad40 and Ad41 could be typed by PCR. To extend the technique of type-specific PCR to other adenovirus serotypes, type-specific primers for Ad1, Ad2, Ad4, Ad5, Ad19 and Ad37 were evaluated. In the present study, 50 stool and 68 eye swab specimens were first tested for the presence of adenoviruses using genus-specific primers. Adenoviruses could be detected in 42 stool and 47 eye swab samples. While the adenovirus-positive stool samples were subsequently typed with primers for Ad2, Ad5, Ad31, Ad40 and Ad41, the positive eye swab specimens were typed with primers for Ad4, Ad8, Ad19 and Ad37. PMID- 9201816 TI - Drugs in pregnancy. Introduction. PMID- 9201815 TI - Characterization of iridovirus IV1 polypeptides: mapping by surface labelling. AB - A comprehensive index of IV1(Tipula iridescent virus, or TIV)-associated polypeptides has been established using enrichments of "empty" and "filled" virions that are considered to be maturation-phase-related. The mapping strategy which involved one- and two-dimensional polyacrylamide gel electrophoresis, silver staining, disulphide bond reduction and 125I iodination of putative surface proteins revealed 103 and 116 polypeptides for empty and filled capsids, respectively. These estimates could be reduced to < or = 70 by reclassing multicharged polypeptides of approximately identical masses as single entitles. At least 10 polypeptides from empty and filled virions were involved in intermolecular sulphhydryl linkages and another 11 species were identified as putative outer shell (surface) polypeptides. These data provide useful criteria for iridovirus classification and identification of candidate polypeptides involved in capsid formation and maturation. PMID- 9201817 TI - Drugs in pregnancy: anticonvulsants. AB - Although 90% of patients using anticonvulsant drugs can expect a favorable pregnancy result, this outcome can be maximized by careful preconceptional, antepartum, and postpartum management. There is no clearcut agreement that any one of the four major drugs used for the treatment of seizure disorders (phenytoin, phenobarbital, valproic acid, and carbamazepine) is more teratogenic than others. Patients should be counseled that the risk of adverse maternal and neonatal outcomes from recurrent seizures during pregnancy is greater than the risk of teratogenicity with anticonvulsant drugs. The lowest possible dose of one of the four front-line agents is recommended for seizure control in pregnancy. The interaction of epilepsy and pregnancy, as well as the possible mechanisms of anticonvulsant teratogenesis, are reviewed. The use and toxicity of individual, commonly used, anticonvulsant drugs are described. Detailed consensus recommendations for preconceptional, antenatal, intrapartum, and postpartum management of patients using anticonvulsant drugs are discussed. PMID- 9201818 TI - Angiotensin converting enzyme inhibitors in pregnancy. AB - Angiotensin converting enzyme (ACE) inhibitors are excellent antihypertensive agents and are becoming widely used as first-line therapy for chronic hypertension in women of reproductive age owing to their efficacy and few side effects. Reports of adverse fetal and neonatal effects from the use of ACE inhibitors in pregnancy in both animal and human studies prompted recommendations against their use in human pregnancy by several authors. This review discusses the mechanism of action of ACE inhibitors and the use of ACE inhibitors in pregnancy both in experimental animals and use in human pregnancy. ACE inhibitors used during pregnancy may have untoward effects on the fetus. Based on reports in the literature, one should avoid starting ACE inhibitors during pregnancy and discontinue them in current users if at all possible. PMID- 9201819 TI - Vitamin A and its congeners. AB - Vitamin A (retinol) is a fat-soluble vitamin that is necessary for cell growth and differentiation. Excess vitamin A has been associated with teratogenic effects in animals and humans. Because vitamin A deficiency is very uncommon in the industrialized world, the current recommendation is that routine vitamin A supplementation is not necessary. If vitamin A supplements are used, they should be limited to less than 5,000 IU per day. Systemic administration of the naturally occurring retinoid tretinoin has been associated with birth defects, fetal resorption, and stillbirths in animals; however, topical use is not associated with increased birth defects and is classified as a category B drug during pregnancy. The synthetic retinoids isotretinoin, etretinate, and etretin are strictly contraindicated during pregnancy (category X) as they have been associated with teratogenic syndromes in humans. In addition, owing to the prolonged elimination half-life of aromatic retinoids, effective contraception should be used for at least 2 years following discontinuation of treatment with these drugs. PMID- 9201820 TI - Immunosuppressant therapy during gestation. AB - Use of immunosuppressants during pregnancy is indicated for anti-rejection therapy in transplantation patients and treatment of autoimmune diseases. Maternal side effects include nephrotoxocity and hepatotoxicity. All immunosuppressant drugs cross the placenta. Immunosuppressant use during the first trimester is not strongly associated with an increased risk of congenital anomalies, although some agents (eg, azathioprine) may be associated with slightly increased frequencies of birth defects. Effects of exposure to this class of drugs during the second and third trimesters affects the fetus' immune system. The result is an infant with a transiently compromised immune system at an increased risk of slightly lower birth weight. Other direct toxic effects of the drugs on the infant's pancreas, liver, and lymphocytes are reported. Certain agents (eg, penicillamine, chloroquine) should be avoided during pregnancy, if possible. However, their use cannot be discontinued during pregnancy given the life-threatening nature of the indication for use of immunosuppressants. PMID- 9201821 TI - Anticoagulants and thrombolytics during pregnancy. AB - Although venous thromboembolism is a rare complication of pregnancy, it is one of the leading causes of maternal mortality. As many as 40% of asymptomatic women with deep venous thrombosis may indeed have a pulmonary embolism. Therefore, pregnant women with thromboembolic disease, a history of thromboembolic disease, or those who are at increased risk of thromboembolism (mechanical cardiac valve prostheses, antithrombin II, or protein C or S deficient) should receive anticoagulant therapy. The choice of anticoagulant therapy in a pregnant woman as well as the dose and duration will depend on the specific condition being treated. Although anticoagulant therapy is beneficial, it is not without risks to both mother and fetus. This article discusses the use of anticoagulants and thrombolytics in pregnant women. PMID- 9201822 TI - Psychotropics. AB - The risks during pregnancy from commonly prescribed psychopharmacological agents are summarized in this article. Psychotropic agents included in this discussion are antidepressants, antianxiety agents, and antipsychotic agents. The focus of this review is to describe the association between these medications and the incidence of congenital malformations. Epidemiological studies, cohort studies, case reports, and animal studies are discussed. PMID- 9201823 TI - [Effects of frame of reference on the judgments of whole-body vibration intensity]. AB - Although the concept of the term 'riding comfort' is ambiguous, in the present paper it means a perceptual experience derived from the vibrational factors of a running railway vehicle. When we regard riding comfort evaluation as a perceptual judgment process, we must consider that what is perceived is dependent not only on the physical properties of the stimuli, but also on the frame of reference. The purpose of the present study is to examine the effect of the frame on the judgments of vibration intensity in the anchoring effect paradigm. Using the four axis vibration apparatus, we conducted experiments for eighty subjects, in which frequencies and lateral accelerations of vibrations were changed. As the result, we found a clear anchoring effect. This suggests that we must take into consideration effects of frame of reference in terms of riding comfort criterion of railway vehicles. PMID- 9201824 TI - [A socio-relational basis of "irrational" cooperation: an experimental study with the selective-play paradigm]. AB - Experimental gaming researchers have recently realized that a larger context is important in which dyadic prisoner's dilemma (PD) relations are embedded. The main purpose of this study was to examine whether people facing a "selective play" situation spontaneously adopt the out-for-tat strategy (OFT), which previous studies found the most effective in such a situation. OFT proscribes to (1) always cooperate, (2) keep playing with the current player as long as he/she cooperates, and (3) desert the partner as soon as he/she stops cooperating. Results of an experiment with 90 students, of groups of six or eight, showed that subjects who faced a high level of social uncertainty in fact spontaneously adopted the strategy, interacting with the same partner as long as he/she cooperated, and exiting from the relation when the partner defected. Subjects who faced a low level of social uncertainty kept only the third part of the strategy, leaving the relation with a defector, but not necessarily keeping interaction with a cooperative partner. PMID- 9201825 TI - [A study in cognitive complexity of the self: an evaluation of the Linville's index]. AB - The purpose of the present study was to evaluate H statistic, proposed by Linville (1985, 1987), as an index for cognitive complexity of the self. Linville asserted that high self-complexity would act as a buffer against life stress or depression. One hundred and eighty-seven undergraduates sorted 40 personality trait adjectives into as many categories as necessary in order to describe themselves. In addition, 126 participants filled out several scales including self-consciousness and esteem. Main findings were as follows: (a) H statistic was not significantly associated with any variable related to the self-ratings, and showed no stress-buffering effect. (b) On the other hand, participants who had high cognitive complexity for the negative aspects of the self, as operationalized by Woolfolk, Novalany, Gara, Allen, and Polino (1995), were low in self-esteem and high in public self-consciousness. The results suggest that cognitive complexity of the negative self may indicate a predisposition for depression or neurosis. (c) Also, women scored significantly higher than men on cognitive complexity of the negative self. PMID- 9201826 TI - [A study of gender difference in student apathy]. AB - This study attempted to construct Male Apathy Inventory (MAI), Female Apathy Inventory (FAI), and Obsession Scale (OS), and investigate the relations among apathy, obsession, self-consciousness, and self-image. Forty-seven original items for MAI and FAI, 13 original items for OS, Sugawara's self-consciousness scale (1984), and Murase and Murase's self-image scale (1966) were administered to 508 university students. Principal component analysis and factor analysis were used to select 24, 32, and 9 items for MAI, FAI, OS, respectively. Students high on OS tended to be higher on private self-consciousness, and male students who were low on both apathy and obsession had the lowest public self-consciousness. Self images of the students high on apathy tended to be negative. Also, high obsession scores seemed beneficial for the adjustment of female students. PMID- 9201827 TI - [A new wave of behavior genetic modeling using covariance structure analysis]. AB - A number of useful methods for analyzing covariance structure have been proposed in the studies of human behavior genetics, reflecting the fact that the behavior genetic studies are one of the main origins of covariance structure model. In this paper, I review recent progress on methodology for behavior genetic studies of twins and families from the standpoint of the structural equation modeling. Especially, genetic ACE (additive genetic, common environment and random environment) model, multivariate ACE model, genetic factor analysis model and twin-parent model are focused upon. This review also discusses how to construct applied structural equation models which are useful for psychological research. PMID- 9201828 TI - To brace or not to brace: the true value of school screening. PMID- 9201829 TI - Viscoelastic relaxation and regional blood flow response to spinal cord compression and decompression. AB - STUDY DESIGN: To better understand the relationships between primary mechanical factors of spinal cord trauma and secondary mechanisms of injury, this study evaluated regional blood flow and somatosensory evoked potential function in an in vivo canine model with controlled velocity spinal cord displacement and real time piston-spinal cord interface pressure feedback. OBJECTIVES: To determine the effect of regional spinal cord blood flow and viscoelastic cord relaxation on recovery of neural conduction, with and without spinal cord decompression. SUMMARY OF BACKGROUND DATA: The relative contribution of mechanical and vascular factors on spinal cord injury remains undefined. METHODS: Twelve beagles were anesthetized and underwent T13 laminectomy. A constant velocity spinal cord compression was applied using a hydraulic loading piston with a subminiature pressure transducer rigidly attached to the spinal column. Spinal cord displacement was stopped when somatosensory evoked potential amplitudes decreased by 50% (maximum compression). Six animals were decompressed 5 minutes after maximum compression and were compared with six animals who had spinal cord displacement maintained for 3 hours and were not decompressed. Regional spinal cord blood flow was measured with a fluorescent microsphere technique. RESULTS: At maximum compression, regional spinal cord blood flow at the injury site fell from 19.0 +/- 1.3 mL/100 g/min to 12.6 +/- 1.0 mL/100 g/min, whereas piston spinal cord interface pressure was 30.5 +/- 1.8 kPa, and cord displacement measured 2.1 +/- 0.1 mm (mean +/- SE). Five minutes after the piston translation was stopped, the spinal cord interface pressure had dissipated 51%, whereas the somatosensory evoked potential amplitudes continued to decrease to 16% of baseline. In the sustained compression group, cord interface pressure relaxed to 13% of maximum within 90 minutes; however, no recovery of somatosensory evoked potential function occurred, and regional spinal cord blood flow remained significantly lower than baseline at 30 and 180 minutes after maximum compression. In the six animals that underwent spinal cord decompression, somatosensory evoked potential function and regional spinal cord blood flow recovered to baseline 30 minutes after maximum compression. CONCLUSIONS: Despite rapid cord relaxation of more than 50% within 5 minutes after maximum compression, somatosensory evoked potential conduction recovered only with early decompression. Spinal cord decompression was associated with an early recovery of regional spinal cord blood flow and somatosensory evoked potential recovery. By 3 hours, spinal cord blood flow was similar in both the compressed and decompressed groups, despite that somatosensory evoked potential recovery occurred only in the decompressed group. PMID- 9201830 TI - Progression of vertebral wedging in an asymmetrically loaded rat tail model. AB - STUDY DESIGN: A rat tail model was used to test the hypothesis that angulation and asymmetric axial compressive loading would lead to vertebral wedging because of asymmetric longitudinal growth in the physes. OBJECTIVES: To study the effect of angulation and asymmetric loading on the progression of spinal curvature in a rat tail model. SUMMARY OF BACKGROUND DATA: Large idiopathic scoliotic curves in children with significant growth remaining are the curves most likely to progress. The mechanism of progression of skeletal deformities is thought to be controlled by the Hueter-Volkmann law, whereby additional axial compression decelerates growth, and reduced axial compression accelerates growth. It has been hypothesized that spinal curvature leads to asymmetric loading transversely along the vertebral growth plate, causing progressive vertebral wedging by means of a vicious cycle. METHODS: Two 32-mm diameter external ring fixators were glued to 0.7-mm pins that had been inserted percutaneously through the eighth and 10th caudal vertebra of 10 6-week-old Sprague-Dawley rats. Calibrated springs and 15 degrees wedges, mounted on stainless steel threaded rods passing through holes distributed around the rings, imposed a 30 degrees Cobb angle and axially compressed the instrumented vertebrae. Fluorochrome labels and radiographs were used to document the progression of vertebral wedging. RESULTS: The wedging initially was entirely in the intervertebral discs, but by 6 weeks the wedging of the discs and vertebrae were approximately equal. Fluorochrome labeling confirmed that the vertebral wedging resulted from asymmetric growth in the physes. CONCLUSIONS: This study shows that vertebrae, when asymmetrically loaded, become wedged. This is consistent with the concept of mechanically provoked progression of scoliotic deformities according to the Hueter-Volkmann law. PMID- 9201831 TI - Role of serotonin for scoliotic deformity in pinealectomized chicken. AB - STUDY DESIGN: The effect of intraperitoneal injection of 5-hydroxytryptophan (5 HTP) versus control in pinealectomized chickens. OBJECTIVE: To find if the serotonin may have some role in the cause of treatment of idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: One of the causes of idiopathic scoliosis is thought to be the disruption of postural reflex. Serotonin has been proposed to have a crucial role in maintaining normal postural muscle tone or postural equilibrium. METHOD: Forty pinealectomized chickens served as controls, and an additional 40 pinealectomized chickens received daily intraperitoneal injections of 5-hydroxy tryptophan, a precursor of serotonin, which can pass through the blood-brain barrier. Spine radiographs were examined to measure the scoliotic deformity. RESULTS: Scoliosis developed in all 40 pinealectomized chickens (control), whereas only 28 chickens in the 5-hydroxytryptophan-treated group (6 in severe, 22 in mild) had scoliosis developed. The remaining 12 chickens grew up with normal spines. Most chickens with mild scoliosis did not have curve progression but continued to have wedged vertebrae. CONCLUSION: Serotonin deficit secondary to a defect of melatonin may have disturbed postural muscle tone or postural equilibrium resulting in scoliosis in pinealectomized chicken. Prevention from the development of scoliosis or its progression in chickens treated with 5 hydroxytryptophan suggests that serotonin may have potential therapeutic value. PMID- 9201832 TI - A comparison between the Boston brace and the Charleston bending brace in adolescent idiopathic scoliosis. AB - STUDY DESIGN: The authors studied 319 patients with adolescent idiopathic scoliosis treated at the same institution with either a Boston brace or a Charleston bending brace. OBJECTIVES: To determine if both orthoses are equally effective in stopping curve progression and preventing the need for surgical correction. SUMMARY OF BACKGROUND DATA: Early reports suggest that the Charleston brace may be comparable to the Boston brace in its effectiveness and that both braces positively influence the natural history of idiopathic scoliosis. METHODS: Skeletally immature (Risser 0, 1, or 2) patients with idiopathic scoliosis who were 10 years old or older at the time of brace prescription, had curves from 25 degrees to 45 degrees, and had no prior treatment were studied retrospectively. All measurements were collected by a single observer, and all patients were followed up to skeletal maturity. RESULTS: The Boston brace is more effective than the Charleston brace, both in preventing curve progression and in avoiding the need for surgery. These findings were most notable for patients with curves of 36 degrees-45 degrees, in whom 83% of the those treated with a Charleston brace had curve progression of more than 5 degrees, compared with 43% of those treated with the Boston brace (p < 0.0001). CONCLUSION: When given the choice between these two orthoses in the treatment of adolescent idiopathic scoliosis, the authors recommend use of the Boston brace. The Charleston brace should be considered only in the treatment of smaller single thoracolumbar or single lumbar curves. PMID- 9201833 TI - Chiari I malformation associated with syringomyelia and scoliosis. AB - STUDY DESIGN: A retrospective review of a series of 12 children who underwent suboccipital foraminotomy and duroplasty for Chiari I malformation. OBJECTIVE: To assess the effects of this surgery on associated syringomyelia and scoliosis. SUMMARY OF BACKGROUND DATA: Suboccipital foraminotomy for the treatment of syringomyelia associated with Chiari I malformation was greatly stimulated by Gardner's hydrodynamic theory, and its results proved to be encouraging. However, several authors reported improvement or stabilization of associated scoliosis after this surgery. METHODS: A retrospective review was conducted on 12 patients who underwent suboccipital foraminotomy for Chiari I malformation associated with syringomyelia. Neurologic Impairment, extent of syringomyelia, and severity of associated spinal deformity were assessed preoperatively and at a 4.5-year average follow-up (range, 2.1-12 years). Anomaly of superficial abdominal reflexes was found in all cases, and para or tetraparesis in three cases. Syringomyelia was of variable localization and extent. Scoliosis was present in 7 cases (greater than 40 degrees in 5 cases). RESULTS: Diminution or complete disappearance of syringomyelia was observed in 11 cases, 3 months to 1 year after surgery. Superficial abdominal reflexes anomaly improved in four cases. Minimal neurologic deficit persisted in one case. Scoliosis improved in one case, remained unchanged in one case, and progressed in the five cases with preoperative severe deformity, requiring instrumentation and fusion. CONCLUSIONS: Improvement of syringomyelia and neurologic deficit, observed with suboccipital foraminotomy, supports the theory that abnormal hydrodynamics of the cerebral spinal fluid is most likely to cause these deficits. PMID- 9201834 TI - Ophthalmic complications after spinal surgery. AB - STUDY DESIGN: A retrospective review of 3450 spinal surgeries was performed. OBJECTIVES: To review ophthalmic complications and their etiologies, as well as treatments and outcomes, in patients who have undergone spinal surgery. SUMMARY OF BACKGROUND DATA: Ophthalmic complications after major spinal reconstructive surgery are rare and have not been adequately addressed in the orthopedic literature. METHODS: In a series of 3450 spinal surgeries at three institutions, the authors identified seven patients (incidence = 0.20%) whose postoperative course was complicated by loss of visual acuity. These perioperative ophthalmic complications included posterior optic nerve ischemia, occipital lobe infarcts, and central retinal vein thrombosis. Operative time, estimated blood loss, and medical history of peripheral vascular, cardiovascular, or ophthalmic disease were obtained from the charts, as were follow-up data. RESULTS: Three patients recovered completely, and one had partial return of visual function. In the remaining three patients, significant visual loss persisted. CONCLUSIONS: The risk of ophthalmic complications with spinal surgery has not been fully appreciated. Because ophthalmic complications in spinal surgery may be reversed with prompt recognition and intervention, it is important for clinicians to be aware of their possible occurrence. PMID- 9201835 TI - Visual loss as a complication of spine surgery. A review of 37 cases. AB - STUDY DESIGN: Thirty-seven patients who experienced visual loss after spine surgery were identified through a survey of the members of the Scoliosis Research Society and a review of the recent literature. OBJECTIVES: Records were reviewed in an attempt to identify preoperative and intraoperative risk factors and to assess the likelihood of recovery. SUMMARY OF BACKGROUND DATA: Postoperative blindness after spine surgery has been documented in case reports or small series. The authors report the largest group of such cases to date and the first to allow conclusions regarding risk and prognosis. METHODS: Letters were sent to members of the Scoliosis Research Society requesting copies of medical records concerning patients who experienced postoperative visual deficits after spine surgery. An additional 10 well-documented recent cases were identified from published reports. RESULTS: Patients with visual loss had a mean age of 46.5 years. Surgery included instrumented posterior fusion in 92% of the cases, with an average operative time of 410 minutes and blood loss of 3500 mL. Most cases had significant intraoperative hypotension, with a mean drop in systolic blood pressure from 130 to 77 mm Hg. However, comparison with a matched group of patients with no visual symptoms showed no differences in the hematocrit or blood pressure values. Visual loss occurred because of ischemic optic neuropathy, retinal artery occlusion, or cerebral ischemia. Eleven cases were bilateral, and 15 patients had complete blindness in at least one eye. Most deficits were permanent. CONCLUSIONS: The authors conclude that blindness after spine surgery is more common than has been recognized previously. Most cases are associated with complex instrumented fusions. PMID- 9201836 TI - The treatment of progressive kyphoscoliosis in camptomelic dysplasia. AB - STUDY DESIGN: This study evaluated the different forms of treatment of camptomelic dysplasia, a rare form of short-limbed dwarfism. OBJECTIVES: To determine the most efficacious form of management of spinal deformities in camptomelic dysplasia. SUMMARY OF BACKGROUND DATA: The literature on treatment of spinal deformities in camptomelic dysplasia is sparse. One report advocates aggressive surgical treatment to prevent curve progression and prevent already compromised respiratory function. METHODS: Eight patients with camptomelic dysplasia and progressive spinal deformity underwent a retrospective chart and radiographic review by an independent observer. Follow-up averaged 3 years and 9 months. RESULTS: Five of eight patients initially were treated with bracing and six of eight patients eventually required surgery. Average initial kyphosis was 114 degrees and scoliosis 61 degrees, compared with 99 degrees kyphosis and 52 degrees scoliosis at follow-up. Complications included pseudarthrosis (50%) and neurologic problems (33%). CONCLUSIONS: The authors advocate anterior/posterior uninstrumented fusion and halo cast immobilization postoperatively to prevent curve progression and avoid the potentially fatal sequelae associated with this disorder. PMID- 9201837 TI - Allograft bone use during instrumentation and fusion in the treatment of adolescent idiopathic scoliosis. AB - STUDY DESIGN: In a retrospective study, 25 patients undergoing posterior spine fusion with allograft bone and Cotrel-Dubousset Instrumentation were assessed regarding the efficacy of allograft bone use. OBJECTIVES: To determine if allograft bone use had deleterious effects regarding fusion rates and maintenance of deformity correction. SUMMARY OF BACKGROUND DATA: Previous studies using allograft bone in adult lumbar spine fusion models have consistently shown poor fusion rates. Studies in the pediatric population have been more favorable but in idiopathic cases have used cast or brace immobilization with Harrington instrumentation. METHODS: Twenty-five skeletally immature patients with is average age of 14 +/- 4 years and an average follow-up of 4 +/- 2 years (minimum of 3 years) were evaluated with anteroposterior, lateral, and oblique radiographs to assess the fusion mass. RESULTS: Preoperative curves averaged 55.5 degrees and immediate correction averaged 58% with an average postoperative curve of 23.2 degrees. Loss of correction at final follow-up was 3.7 degrees. No pseudarthroses were identified clinically or radiographically. CONCLUSIONS: Allograft bone use in the pediatric patient with idiopathic scoliosis undergoing rigid segmental instrumentation dependably results in fusion with good maintenance of correction. PMID- 9201838 TI - Posterior arthrodesis in the skeletally immature patient. Assessing the risk for crankshaft: is an open triradiate cartilage the answer? AB - STUDY DESIGN: Thirty-three skeletally immature patients younger than 12 years of age and having posterior arthrodesis and evidence of solid posterior fusion without "adding on" were retrospectively reviewed. All patients had a minimum of 5 years of follow-up. OBJECTIVES: To ascertain factors associated with crankshaft and to determine how accurate a marker the triradiate cartilage was. SUMMARY OF DATA: All patients had Risser Stage 0 curves and all of the girls were premenarchal preoperatively. The average age was 9 years 3 months (range, 2 years 11 years 11 months). Preoperative diagnoses consisted of 14 idiopathic, 11 congenital, five dysplastic, and three neuromuscular etiologies. METHODS: Preoperatively, within 3 months after surgery, and at 2-year, 5-year, and final postoperative follow-up, the following radiographic parameters were reviewed: coronal Cobb, apical vertebral rotation, apical vertebral translation, rib vertebral angle difference, and trunkshift. RESULTS: The triradiate cartilage was open in 24 patients at the time of operation. Of those 24, only nine (37.5%) had documented proof of crankshaft. Patients with closed triradiate cartilage had no significant postoperative increase in radiographic parameters (0 of 9). The subgroup of patients with idiopathic scoliosis had an average age of 11 years 3 months (range, 9 years 2 months-11 years 11 months). Five of 14 patients had an open triradiate cartilage. All were followed up to skeletal maturity. None had significant progression in postoperative radiographic parameters. CONCLUSION: This study did not find an open triradiate cartilage to be an absolute prognostic indicator for the occurrence of crankshaft. Additional refinement of markers of maturity are needed to determine who requires anterior arthrodesis. PMID- 9201839 TI - Prediction of the crankshaft phenomenon by peak height velocity. AB - STUDY DESIGN: Retrospective review. OBJECTIVES: To evaluate the relation of the peak height velocity with the occurrence of the crankshaft phenomenon after posterior arthrodesis and instrumentation in idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Although patients with closed triradiate cartilages are unlikely to exhibit the crankshaft phenomenon after a posterior spinal fusion and instrumentation, open triradiate cartilages do not necessitate that crankshafting will occur. Less than half of patients with idiopathic scoliosis and open triradiate cartilages will exhibit the crankshaft phenomenon. METHODS: The authors reviewed 43 patients with idiopathic scoliosis who were Risser 0 at the time of posterior spinal fusion. Twenty-three patients had open triradiate cartilages and twenty had closed. The timing of peak height velocity was identified. RESULTS: All patients with closed triradiate cartilages were beyond their peak height velocity at the time of surgery. Among those with open triradiate cartilages, 8 were operated on before or during their peak and 15 were operated on afterward. All patients fused before or during the peak crankshafted. Two of the fifteen patients fused after the peak crankshafted. In one, it was low grade. In the other, it appears that the fusion blunted the peak height velocity to a point at which it was unidentifiable. CONCLUSIONS: In patients with open triradiate cartilages, surgery performed before or during the peak height velocity is a strong predictor of the crankshaft phenomenon, and later surgery is a strong negative predictor of the crankshafting (P = 0.000009). Isolated posterior fusion before the height velocity decelerates results in the crankshaft phenomenon, whereas fusion during the deceleration phase does not. PMID- 9201840 TI - Anterior correction of thoracic scoliosis with Kaneda anterior spinal system. A preliminary report. AB - STUDY DESIGN: Analysis of the clinical results of 20 patients with thoracic scoliosis treated by anterior procedure with Kaneda anterior spinal system. OBJECTIVES: To evaluate the efficacy of the anterior surgical correction procedure with a new anterior instrumentation in thoracic scoliosis. SUMMARY OF BACKGROUND DATA: Posterior correction and fusion with posterior instrumentation has been a main component of the surgical management of thoracic scoliosis. However, to the best of the authors' knowledge, no clinical results of anterior instrumentation surgery for thoracic scoliosis have been published in the English literature. METHODS: Anterior correction and fusion using Kaneda anterior spinal system was performed in 20 patients with thoracic scoliosis (3 patients with King Type II curve, 13 with Type III, and 4 with Type IV). The average follow-up was 3 years, with a range of 2 years, 3 months to 4 years, 1 month. There were 1B patients with idiopathic scoliosis (13 adolescents and 5 adults) and 2 patients with a single thoracic curve caused by other etiologies. All patients had correction of scoliosis by fusion within the major thoracic curve. Radiographic evaluations were performed to analyze frontal, sagittal, and rotational deformities of the spine. RESULTS: The average correction rate of scoliosis was 71%. Above the instrumented levels, the correction rate was 75%. Preoperative kyphosis of the instrumented levels of 7 degrees was corrected to 14 degrees of kyphosis. The trunk shift was improved from 17 mm before surgery to 9 mm at final follow-up evaluation. The average improvement of the tilt-angle in the lower and vertebra of fusion was 81%, and was 83% in the stable vertebra. Apical vertebral rotation showed correction rate of 15% in patients without performing resection of the rib head joints and rod rotation maneuver (n = 6). However, the correction rate was improved to 58% after introduction of the technique discussed (n = 14). The angle of tangential rib deformity (rib hump) showed a correction rate of 50%. There was 1.2 degrees of frontal plane and 1.0 degree of sagittal plane correction loss within the instrumented area at final follow-up evaluation. At final follow-up, nonunion at the uppermost segment of the fusion range developed in one patient, and decompensation in the lumbar spine was observed in one patient with Type II curve. CONCLUSIONS: Anterior correction with Kaneda anterior spinal system provides excellent correction of the frontal curvature and sagittal alignment by fusing within the range of the major curve, without a significant loss of correction and implant failure. Rigid rotational deformity of the thoracic scoliosis is effectively corrected by resection of the rib head joints and rod rotation maneuver. However, too much correction of the thoracic curve should be avoided, to prevent decompensation of the lumbar curve, especially in Type II curves. PMID- 9201841 TI - Lumbar pedicle screws versus hooks. Results in double major curves in adolescent idiopathic scoliosis. AB - STUDY DESIGN: A retrospective assessment of the effectiveness of lumbar pedicle screws versus laminar hooks in lumbar curve correction with double major curves in adolescent idiopathic scoliosis. OBJECTIVE: To determine if pedicle screw fixation of the lumbar spine has any advantage compared with multiple laminar hook instrumentation in the treatment of double major curves in adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: Although hooks have been used most commonly, pedicle screws may offer advantages in correction and maintenance of reduction of the lumbar curve in adolescent idiopathic scoliosis. METHODS: A consecutive series of 39 patients with double major curves underwent thoracic and lumbar instrumentation by a single surgeon. Lumbar pedicle screws and hooks were used in 20 patients (Group S) and in 19 patients only lumbar hooks were used (Group H). Thoracic Cotrel-Dubousset instrumentation with hooks was the same in both groups. Preoperative age, gender, bracing, and Cobb angles were similar in both groups. Preoperative, 1-month postoperative, and latest follow-up standing posteroanterior and lateral spine radiographs were blinded to the surgeon and lumbar instrumentation covered to hide its identity. Measurements included Cobb angles, preoperative flexibility, lumbar and thoracic apical vertebral deviation, and reduction of lateral tilt and lateral displacement of the first free lumbar vertebra below the instrumentation. Percent correction, maintenance of correction at follow-up, and total levels fused were calculated. RESULTS: The mean follow-up was 3.5 years (range, 2-8 years), which was similar for Groups H and S. Pedicle screws appear to offer some advantage in lumbar curve correction, maintenance of correction, and correction of the uninstrumented spine below the fusion when compared with the use of hooks alone. Horizontalization of the first free lumbar vertebra below the instrumentation percent correction of tilt: 62% screws vs. 11% hooks; P = 0.0003), residual tilt (8 degrees screws vs. 17 degrees hooks; P = 0.004), and loss of horizontalization at follow-up (5% screws vs. 26% hooks) were dramatically better for the group using screws. Lumbar curve correction (72% screws vs. 60% hooks; P = 0.026), loss of lumbar curve correction (5% screws vs. 13% hooks), and correction of lateral apical vertebral deviation (2.2-cm screws vs. 1.5-cm hooks or 63% vs. 31%; P = 0.013) were better when screws were used. There was no significant difference in loss of correction of the thoracic curves (35% vs. 37%) or any difference in loss of correction of lateral displacement of the thoracic apical vertebra (12% vs. 14%). There was no difference in total levels fused, operative blood loss, operative time, or ultimate patient outcome. No patients in either group had spinal imbalance at latest follow-up. There were no complications related to pedicle screw placement. Two cases of transient postoperative superior mesenteric artery syndrome (duodenal obstruction by the superior mesenteric artery) in the pedicle screw group are attributed to acute correction of the lumbar scoliosis and thoracolumbar kyphosis with resultant lordosis at the thoracolumbar junction. CONCLUSIONS: Lumbar pedicle screws may offer greater lumbar curve correction, better maintenance of correction, and greater correction of the uninstrumented spine below double major curves. No complications were associated with the placement of pedicle screws. PMID- 9201842 TI - Does scoliosis have a psychological impact and does gender make a difference? AB - STUDY DESIGN: A population-based case-control study, we identified adolescents with and without scoliosis in Minnesota who were 12 through 18 years of age. Matched control subjects were randomly selected from school children who did not have scoliosis or any other condition. Information on scoliosis was obtained by a self-administered questionnaire, the Adolescent Health Survey. Collected on more than 75,000 school age adolescents, with established validity and reliability, a secondary analysis of adolescents with scoliosis was performed as compared with a normative peer group. OBJECTIVE: To describe and characterize the psychosocial impact of scoliosis on the areas of peer relations, body image, and health compromising behavior, such as suicidal thought and alcohol consumption. SUMMARY OF BACKGROUND DATA: The impact of adolescent idiopathic scoliosis has not been assessed using generic health status measures appropriate for adolescents. Previous studies have concentrated on the health status of adults by measuring work status, marriage status, and other adult measures. The purpose of this study was to study the health status of patients with adolescent idiopathic scoliosis, using the Adolescent Health Survey, a generic health status measure with established validity and reliability. METHODS: Body image, peer relations, social and high-risk behavior, and comparative health were assessed to determine if scoliosis was an independent risk factor and to determine if scoliosis was associated with these psychosocial issues. RESULTS: Six hundred eighty-five cases of scoliosis were identified from the 34,706 adolescents. The prevalence was 1.97%. Of the 685 adolescents with scoliosis and their control subjects, the adjusted odds ratio for having suicidal thought among adolescent with scoliosis, compared to adolescents without scoliosis, was 1.40 (P value of 0.04) after adjustment for race, gender, socioeconomic status, and age. The adjusted odds ratio for having feelings about poor body development among adolescents with scoliosis was 1.82 (P value 0.001) compared with adolescents without scoliosis after adjustment for race, gender, socioeconomic status, and age. Scoliosis was an independent risk factor for suicidal thought, worry and concern over body development, and peer interactions after adjustment. CONCLUSION: Scoliosis is a significant risk factor for psychosocial issues and health-compromising behavior. Gender differences exist in male and female adolescents with scoliosis. PMID- 9201843 TI - Evaluation of motor function during thoracic and thoracolumbar spinal surgery based on motor-evoked potentials using train spinal stimulation. AB - STUDY DESIGN: Using compound muscle action potentials after train spinal stimulation, intraoperative motor functional monitoring was performed during thoracic and thoracolumbar spinal surgery. OBJECTIVES: This study was designed to clarify the clinical usefulness of train spinal stimulation and to determine the critical point of compound muscle action potential change at which neurologic injury during surgery occurs. SUMMARY OF BACKGROUND DATA: In 1995 the authors reported that train spinal stimulation allows for the recording of compound muscle action potentials, even in animals and humans under general anesthesia. The facilitative effect of train stimulation overcomes the suppressive effects of anesthetics and allows potentials to pass through synapses, thereby enabling a reliable recording of lower extremity compound muscle action potential. METHODS: Multisegmental recording of compound muscle action potentials after train spinal stimulation was conducted on 34 patients Undergoing surgical treatment for thoracic or thoracolumbar lesions. During surgery, train stimuli (5 pulse, Interstimular Interval: 1 ms) were administered using an epidural electrode introduced transcutaneously. Compound muscle action potentials were recorded from a total of 128 muscles. Anesthesia was maintained using fentanyl and propofol or nitrous oxide with or without isoflurane. Muscle relaxation was attained mainly by controlled infusion of vecuronium bromide. The percent occurrence of recordable compound muscle action potentials was determined, and the potential changes were correlated with changes in muscle strength. RESULTS: Compound muscle action potentials could be recorded from at least one muscle in 94% of the patients, even in most patients with severe motor dysfunction. The compound muscle action potential changes before and after surgical maneuver were divided into four grades. All compound muscle action potential changes in deteriorated muscles belonged to Grade 2 (a 10% latency delay) or Grade 3 (disappearance). CONCLUSIONS: The success rate in obtaining muscle potentials was greatly enhanced when all of the following methods were used: train spinal stimulation, anesthetic with weak suppressive effect, multiple muscle recording, and percutaneous introduction of epidural electrode. The critical point of compound muscle action potential change should be defined as a 10% latency delay or disappearance. Multisegmental muscle potential after train spinal stimulation is the most appropriate method for thoracic and thoracolumbar spinal surgery. PMID- 9201844 TI - Use of intravenous Premarin to decrease postoperative blood loss after pediatric scoliosis surgery. AB - STUDY DESIGN: The hemostatic effect of conjugated estrogens (Premarin, Wyeth Ayerst Laboratories, Philadelphia, PA) has been reported in the literature, but no study has investigated its effectiveness in decreasing blood loss in pediatric spinal procedures. OBJECTIVE: To determine the effectiveness of intravenously administered Premarin in the immediate postoperative period in decreasing Hemovac drainage postoperatively. METHODS: Sixty-four adolescent patients undergoing spinal surgery, both posterior and anterior/posterior procedures, were studied. Thirty-two patients served as control subjects. The other 32 received Premarin, 1 mg/kg intravenously in the immediate postoperative period. Hemovac drainage was measured during a period of 48 hours. RESULTS: In measuring Hemovac drainage for 48 hours in both groups, there was a 37% decrease in the volume of the postoperative drainage (mL/kg) in the patients receiving Premarin postoperatively. Measurement of adjusted volumes (mL/kg) revealed a statistically significant decrease in postoperative bleeding after administration of Premarin. CONCLUSIONS: Administration of Premarin resulted in a 37% decrease in the volume of postoperative drainage (mL/kg) without complications. Premarin appears to be effective in decreasing postoperative blood loss in a pediatric postoperative spinal population. PMID- 9201845 TI - Anterior release and fusion in pediatric spinal deformity. A comparison of early outcome and cost of thoracoscopic and open thoracotomy approaches. AB - STUDY DESIGN: A consecutive series of patients undergoing thoracoscopic anterior spinal release and fusion for scoliosis or kyphosis was compared with a consecutive series of patients treated with an open thoracotomy approach. OBJECTIVES: To compare the early clinical results, costs, and charges of performing an anterior thoracic spinal release and fusion with the two approaches. SUMMARY OF BACKGROUND DATA: The thoracoscopic approach to the spine is gaining acceptance, yet there are little data comparing the technique with standard open methods for the treatment of spinal deformity. METHODS: The first 14 thoracoscopic cases performed at the authors' hospital were compared with 18 open thoracotomy cases treated during the previous 12-month period. In each case the discs were excised and bone grafted before performing a posterior fusion. The early clinical outcomes and the hospital charges/costs were analyzed. RESULTS: The percent curve correction was similar between the thoracoscopic and open methods: scoliosis 56% and 60%, respectively; kyphosis, 88% and 94%, respectively. The blood loss and complication rates were similar between the two groups; however, the chest tube output was greater in the thoracoscopic group. The length of hospital stay was not reduced, and the cost of the open procedure is 29% less than the thoracoscopic approach. The minimally invasive thoracoscopic approach avoids cutting the chest/shoulder musculature, greatly decreasing the morbidity of anterior spinal surgery. CONCLUSIONS: The thoracoscopic technique is a safe and effective alternative to open thoracotomy in the approach to the anterior thoracic spine for the treatment of pediatric and adolescent spinal deformity. PMID- 9201847 TI - Some notable American spine surgeons of the 19th century. AB - In the 19th century, American physicians were drawn to the renowned medical centers of Europe, beacons of excellence in all areas of clinical science, to complement and supplement medical education and training. Such Old World master surgeons as Victor Horsley and William Macewen constantly reported novel and innovative operative procedures, including a significant number of operations on the spine and spinal cord. In this same era, a number of American surgeons made important contributions to the emerging specialty of spine surgery. The lives and work of Alban Smith, Berthold Hadra, DeForest Willard, and Robert Abbe are submitted in this report as examples. PMID- 9201846 TI - Laparoscopic assisted fusion of the lumbosacral spine. A biomechanical and histologic analysis of the open versus laparoscopic technique in an animal model. AB - STUDY DESIGN: An animal model for laparoscopic lumbosacral fusion. OBJECTIVES: To compare the biomechanical and histologic results of open to laparoscopic lumbosacral discectomy and fusion in an animal model. BACKGROUND DATA: Early clinical reports of laparoscopic lumbosacral fusions are encouraging, but animal experiments have not been reported. METHODS: Ten pigs (50-80 kg) were divided into two groups. Group 1 underwent an open anterior lumbosacral discectomy and fusion at L7-S1 using autologous bone graft and a titanium MOSS (DePuy Motech) cage. Group 2 was identical to Group 1 except that a laparoscopic technique was used. The animals were killed at 3 months, and the lumbosacral spines were harvested for biomechanical and histologic testing. RESULTS: Estimated blood loss and average length of operation, respectively, for the two groups were: Group 1, 50 mL, 2 hours 50 minutes; and Group 2, 40 mL, 3 hours 40 minutes. There were no perioperative or postoperative complications in either group. Motion analysis results showed less motion in lateral bending, flexion, and extension than in the intact specimen in both groups. Tensile testing showed that the stiffness was significantly greater in the open group than in the laparoscopic group (P < 0.004). Histologic examination showed a less extensive discectomy and less bone growth in the implant in the laparoscopic group. Inadequate decortication of end plates occurred in two animals who underwent laparoscopy. CONCLUSIONS: Although lumbosacral discectomy and implant insertion can be performed using the laparoscopic technique, the construct may not have the same biomechanical strength as that attained with the open procedure. Laparoscopic-assisted lumbosacral fusion surgery requires additional investigation before it is widely used in clinical situations. PMID- 9201848 TI - Application of polyvinyl alcohol hydrogel membrane as anti-adhesive interposition after spinal surgery. PMID- 9201849 TI - The outcome of posterolateral fusion in highly selected patients with discogenic low back pain. PMID- 9201850 TI - Chronic cervical zygapophysial joint pain after whiplash--a placebo-controlled prevalence study. PMID- 9201851 TI - Functional outcome of thoracolumbar burst fractures managed with hyperextension casting or bracing and early mobilization. PMID- 9201852 TI - Experimental spinal fusion with recombinant human bone morphogenetic protein-2 without decortication of osseous elements. AB - STUDY DESIGN: L4-L5 intertransverse process fusions were produced with 58 micrograms, 230 micrograms, or 920 micrograms of recombinant human bone morphogenetic protein-2 in 20 dogs. Eleven had traditional decortication of posterior elements before insertion of the implant. Nine were left undecorticated. All animals were evaluated 3 months after surgery. OBJECTIVES: To determine whether decortication is a prerequisite for successful fusion in the presence of osteoinductive proteins such as bone morphogenetic protein-2. SUMMARY OF BACKGROUND DATA: Recombinant osteoinductive proteins can induce de novo bone in ectopic soft-tissue sites in the absence of bone marrow elements. Traditional methods for achieving spinal fusion rely on exposure of bone marrow through decortication to facilitate osteogenesis. It is hypothesized that the presence of an implanted osteoinductive protein obviates the need for exposure and release of host inductive factors. METHODS: Recombinant human bone morphogenetic protein-2 induced intertransverse process fusions were performed with and without decortication. Fusion sites were evaluated by computed tomography imaging, high resolution radiography, manual testing, mechanical testing, and histologic analysis. RESULTS: One hundred percent of decorticated spines and 89% of undecorticated spines were clinically fused by 3 months. Ninety-one percent of decorticated spines and 78% of undecorticated specimens exhibited bilateral transverse process osseous bridging. The only spines that failed to achieve solid bilateral arthrodesis were in the lowest dose group. With the higher two doses, there was histologic evidence of osseous continuity between the fusion mass and undecorticated transverse processes. CONCLUSIONS: There were no statistical differences in clinical and radiographic fusion rates between decorticated and undecorticated sites. With higher doses of recombinant human bone morphoganetic protein-2, there was little histologic distinction between fusions in decorticated versus undecorticated spines. PMID- 9201853 TI - Strength and stability of posterior lumbar interbody fusion. Comparison of titanium fiber mesh implant and tricortical bone graft. AB - STUDY DESIGN: A paired comparison was done of the bending flexibility and compression strength of tricortical bone graft and titanium fiber mesh implants in a human cadaver model of posterior lumbar interbody fusion. OBJECTIVES: To test the hypothesis that a titanium fiber mesh implant and a tricortical bone graft provide adequate and equal mechanical strength and stability in posterior lumbar interbody fusion constructs. SUMMARY OF BACKGROUND DATA: Although studies of posterior lumbar interbody fusion constructs have been performed, the authors are unaware of any study in which the strength and stability of a titanium fiber mesh implant are compared with those of tricortical bone graft for posterior lumbar interbody fusion in the human cadaver lumbar spine. METHODS: Changes in neutral zone and range of motion were measured in a bending flexibility test before and after placement of posterior lumbar interbody fusion constructs. Tricortical bone graft and titanium fiber mesh implant construct stability than were compared in a paired analysis. The constructs than were loaded to failure to evaluate construct strength as a function of graft material and bone mineral density. RESULTS: The posterior lumbar interbody fusion procedure produced statistically significant decreases in neutral zone when compared with the intact spine. No statistically significant differences in neutral zone, range of motion, or strength were detected between the two implants. Construct strength correlated strongly with bone mineral density. CONCLUSIONS: Posterior lumbar interbody fusion procedures result in equal or improved acute stability for titanium fiber mesh implants and tricortical bone graft implants when used without additional posterior stabilization. PMID- 9201854 TI - Intermediate-term outcome of cervical spinal cord-injured patients older than 50 years of age. AB - STUDY DESIGN: This study retrospectively reviewed the intermediate-term clinical outcome of patients who were 50 years of age or older at the time they experienced their cervical spinal cord injury. OBJECTIVES: To establish reasonable expectations for the functional outcome in the older patient with cervical spinal cord injury. BACKGROUND DATA: The long-term morbidity and mortality of large groups of patients with spinal cord injury have been reported. The specific functional ability, disposition, morbidity, and mortality of this group of older patients injured after 50 years of age, however, have been less well defined. METHODS: Forty-one consecutive patients older than 50 years of age at the time of cervical cord injury were studied, and functional abilities, independence, need for assistance in activities of daily living, disposition, morbidity, and mortality were assessed. All patients had more than 2 years of follow-up examinations (mean, 5.5 years) by the same spine injury service. RESULTS: There were 13 complete and 28 incomplete cervical cord lesions. The mean age of the patients at follow-up examination was 67.5 years. The average follow up period was 5.5 years after injury. None of the patients with complete cord injury improved, and all required extensive care. Twenty-one (80%) of 26 of the patients with incomplete cord injury were able to ambulate with some assistance. Nineteen of 26 patients had independent or near-independent abilities with activities of daily living. Twenty (77%) of 26 were able to return home. All patients with complete cord injury (13 of 13) had died by the time of the follow up visit. Seventy-seven percent (10 of 13) of this patient group had died within the first year. Those surviving lived an average of 3.5 years after their injury. Fourteen of 28 patients with incomplete cord injury (50%) had died by the time of the follow-up visit. Six (43%) of the 14 deaths were attributed to complications of their spinal cord injury. CONCLUSION: The functional outcome of the person older than 50 years with a complete cervical cord injury is poor. Of the 14% who survived the first year, all required extensive attendant care, and no neurologic improvement was seen. The patient with an incomplete cord injury has an overall good outcome regarding ambulation and returning to home. PMID- 9201855 TI - Efficacy of five cervical orthoses in restricting cervical motion. A comparison study. AB - STUDY DESIGN: Twenty volunteers, 10 men and 10 women, with clinically and radiographically normal cervical spines were studied. OBJECTIVES: To evaluate the effectiveness of five cervical orthoses in their ability to restrict cervical motion in flexion, extension, lateral tilt, rotation, and intervertebral motion. SUMMARY OF BACKGROUND DATA: The five cervical orthoses evaluated were the Philadelphia collar (Philadelphia Collar Co., Philadelphia, PA), Aspen (International Healthcare Devices, Long Beach, CA), Stifneck (Laerdal, Armonk, NY), Miami J (Jerome Medical, Moorestown, NJ), and NecLoc (Jerome Medical, Moorestown, NJ) orthoses. Together these five orthoses comprise 80% of the rigid cervical and extrication devices in current use. METHODS: The normal and unrestricted ranges of active cervical motion in flexion, extension, and lateral tilt were measured in each subject and compared with the motion permitted in each of the five cervical orthoses. Lateral radiographs of the cervical spine in the neutral position and at maximum flexion and extension were obtained. Measurements of flexion, extension, and combined flexion-extension were determined for the cervical spine as a whole as measured from the occiput to the seventh cervical vertebra and at each intervertebral cervical level. Lateral tilt was measured on an anteroposterior radiograph at the extreme of motion. Rotation was measured using a compass goniometer. Each volunteer served as his own control for the radiographic and goniometric measurements. RESULTS AND CONCLUSION: The NecLoc cervical orthosis demonstrated statistically superior restriction of cervical motion in flexion, extension, rotation, and lateral tilt in comparison with the other four orthoses studied in healthy volunteers. The Miami J cervical orthosis was the next most restrictive orthosis and was superior to the Philadelphia Collar and Aspen orthosis in all parameters of motion. PMID- 9201856 TI - Clinical, radiographic, and kinematic results from an adjustable four-pad halovest. AB - STUDY DESIGN: Charts and radiographs of all patients treated with this halovest at one university hospital were reviewed retrospectively. OBJECTIVES: To describe the outcomes from an adjustable four-pad halovest and to compare them with those from standard halovests, as previously published. SUMMARY OF BACKGROUND DATA: With standard halovests, there can be cervical motion up to 70% of normal values, substantial loads between the halo and vest, and complications of pin loosening, pin infections, and scapular pressure sores. The four-pad vest reduces halovest loads and vest-torso motions. METHODS: The four-pad vest has four independently adjustable pads that completely avoid contact with the scapula, clavicle, and abdomen. Clinical records were analyzed to determine the incidences of halo pin loosening, pressure sores, injury or surgical site nonunion, and loss of cervical alignment. Lateral radiographs were taken with the patient in the upright and supine positions at various times to determine intervertebral rotations (flexion extension). RESULTS: The clinical results with the four-pad vest were at least as good as those for standard vests. Scapular pressure sores were prevented completely by the absence of vest-scapula contact. Kyphosis did not increase significantly with time. The mean segmental rotations were all 3 degrees or less and showed a smoothly decreasing pattern from C1-C2 to C6-C7. The value at Oc-C1 was opposite to that at C1-C2 and is the subject of further analysis. CONCLUSIONS: The rotations occurring with the four-pad vest are less than or equal to those occurring with standard vests, for overall cervical rotation and for individual motion segment rotations. This is consistent with the smaller halovest forces seen with this vest. Prospective, comparative testing will assess the clinical significance of these findings. PMID- 9201858 TI - The effect of airway maneuvers on the unstable C1-C2 segment. A cadaver study. AB - STUDY DESIGN: This is a cadaver study in which video fluoroscopy is used to measure motion of the unstable spine at C1-C2 during intubation maneuvers. OBJECTIVES: To quantify the amount of motion that occurs at an unstable C1-C2 spinal segment during the use of various intubation techniques using a cadaver model. SUMMARY OF BACKGROUND DATA: In previous work by the authors, a methodology and measurements for the unstable C5-C6 segment in a cadaver model were developed. These studies showed that the most motion was created by a chin lift and jaw thrust and that oral techniques created more motion than nasal intubation. The potential motion that occurs during intubation with instability at C1-C2 is yet unstudied. Therefore, a study to determine the effects of intubation on the spine with an unstable C1-C2 segment was designed. METHODS: Six human cadavers were used for the study. Measurements before and after transoral osteotomy of the odontoid were performed using video fluoroscopy. Pre-intubation maneuvers and oral and nasal intubation were studied. RESULTS: Oral intubation and nasal intubation caused similar diminution of space available for the cord. Chin lift and jaw thrust caused a larger diminution of space available for the cord than either nasal or oral intubation techniques. CONCLUSIONS: Although nasal intubation is the accepted procedure for intubation of the unstable spine, nasal and oral intubation seemed to have the same ability to narrow the space available for the cord in the model in this study. Great care should be taken while performing the chin lift/jaw thrust maneuvers in preparation for intubation, because these pre-intubation techniques caused the most motion and hence narrowed the space available for the cord in the unstable cervical spine. PMID- 9201857 TI - Occipitocervical fusion with C1 laminectomy in children. AB - STUDY DESIGN: Eight children in whom atlantoaxial dislocation had developed underwent occipitocervical fusion using a rectangular rod. The postoperative results are presented, and the postoperative growth and deformation of the cervical spine were determined radiographically. OBJECTIVES: To investigate in a relatively long-term follow-up study whether occipitocervical fusion affects the growth of the cervical spine and induces spinal deformation. SUMMARY OF BACKGROUND DATA: It has been reported that children who have undergone C1-C2 posterior fusion are likely to develop abnormal curvature or deformation of the cervical spine as a result of a disturbance of growth of the fused vertebrae. There have been no studies, however, to confirm that these changes occur after occipitocervical fusion in children. METHODS: The subjects were one boy and seven girls who had undergone occipitocervical posterior fusion during childhood. The average age at the time of surgery was 8.3 years, and the average follow-up period was 5.9 years. The following were assessed radiographically: redislocation of the atlas, bone union, changes in the curvature of the cervical spine, the height and width of the vertebral bodies, and the anteroposterior diameter of the spinal canal. RESULTS: Solid bone union was achieved in all patients with maintenance of the reduced position at the time of surgery. None of the patients exhibited abnormal curvature of the cervical spine. The rate of increase in height of the C2 vertebral body was significantly less than that of vertebral bodies below C3. The rate of increase in width of the vertebral body and the anteroposterior diameter of the spinal canal of the C2 vertebral body and vertebral bodies below C3 did not differ significantly. CONCLUSIONS: Occipitocervical fusion with a rectangular rod is useful for treating atlantoaxial dislocation in children and yields excellent results because of the firm internal fixation it achieves. This surgery induced no apparent postoperative spinal deformations. PMID- 9201859 TI - The long-term follow-up of patients with Klippel-Feil syndrome and congenital scoliosis. AB - STUDY DESIGN: This study evaluated the long-term results of Klippel-Feil syndrome in patients with congenital scoliosis. OBJECTIVES: To determine the incidence of cervical and cervical-related symptoms of patients who have Klippel-Feil syndrome associated with congenital scoliosis. SUMMARY OF BACKGROUND DATA: Many authors have described the association of Klippel-Feil syndrome and congenital scoliosis. In this population of patients, cervical lesions often are discovered incidentally. The significance of these lesions is unknown. METHODS: Thirty-two patients with congenital scoliosis and Klippel-Feil syndrome were observed for more than 10 years. They were questioned specifically about cervical and cervical related symptoms. All patients had sequential cervical radiographs and physical examinations. RESULTS: Despite rather dramatic radiographic appearances, only seven (22%) of the 32 patients had cervical or cervical-related symptoms, with two patients requiring surgery for their cervical lesions. The extent of the deformities and the average number of cervical vertebrae fused and cervical fusion-patterns were statistically similar between the symptomatic and asymptomatic groups. Patients fused to the cervicothoracic junction for management of their deformities had a significantly increased incidence of cervical symptoms. Also, patients with congenital stenosis had a significantly greater incidence of upper extremity pain. CONCLUSIONS: Only a small number of patients with Klippel-Feil syndrome and congenital scoliosis developed cervical symptoms. No fusion pattern that placed the patient at greater risk for developing symptoms could be identified. Factors that did lead to a greater incidence of cervical symptoms were fusion to the cervicothoracic junction and congenital cervical stenosis. PMID- 9201860 TI - Early detection of progression in adolescent idiopathic scoliosis by measurement of changes in back shape with the Integrated Shape Imaging System scanner. AB - STUDY DESIGN: A retrospective study of 78 patients with right thoracic idiopathic scoliosis was done. OBJECTIVES: To evaluate the reliability of the integrated Shape Imaging System scan (Oxford Metrics Ltd, Oxford, UK) in detecting progression of scoliosis and the use of back shape data in predicting scoliosis progression. SUMMARY OF BACKGROUND DATA: At first presentation and every 3-6 months during the follow-up period, all patients underwent integrated Shape Imaging System scans and radiographic examinations, from which the Cobb angle was measured. The follow-up period was 18-49 months (mean = 31.4 months). METHODS: Patients were divided into three groups according to the severity and progression of the Cobb angle. The spinal fusion, brace, and observation groups were compared using analysis of variance and the student's t test to detect significant differences among groups in the progression of deformity as measured by the integrated Shape Imaging System parameters and the Cobb angle. RESULTS: Three of the Integrated Shape Imaging System parameters detected significant progression in the spinal fusion group 1 year earlier than the Cobb angle. Only one of the Integrated Shape Imaging System parameters detected a significant difference in progression between the brace and observation groups. CONCLUSIONS: The Integrated Shape Imaging System technique demonstrated significant changes in this group of patients with progressive scoliosis. Serial measurements of back surface shape, particularly the size of the rib hump, may be predictive of progression. Serial Integrated Shape Imaging System scanning has advantages over serial radiography in the management of idiopathic scoliosis in addition to the avoidance of exposure to ionizing radiation. PMID- 9201861 TI - The effect of a plaster cast on lumbosacral joint motion. An in vivo assessment with precision motion analysis system. AB - STUDY DESIGN: This study was conducted to assess the effect of a plaster cast on the mobility of the lumbosacral joint in 10 patients with chronic low back pain. During static and dynamic exercises, movements between the proximal vertebra (L4 or L5) and the sacrum were registered in 10 patients without a support and after the application of a plaster cast, and with and without unilateral hip immobilization, respectively. OBJECTIVES: To investigate whether plaster casts actually immobilize the lumbosacral joint. SUMMARY OF BACKGROUND DATA: The presumed stabilizing effect of a lumbar orthosis on the lumbosacral joint has been the subject of many studies in the past years, and contradictory reports have been published. METHODS: The measurements were performed by means of Precision Motion Analysis System, an optoelectronic three-dimensional motion analysis system using infrared light. The patients were asked to perform maximal spinal flexion to extension, maximal pelvic tilt (static test conditions), and to walk within the measurement volume (dynamic test condition). This procedure was repeated with the patients wearing a plaster cast with and without unilateral hip fixation. Mobility was expressed in translations and rotations around three axes. For statistical analysis, repeated measurements two-way analysis of variance was used. RESULTS: Considerable rotations were found only in the sagittal plane. Both plaster casts appeared to decrease mobility during the static test conditions. During the dynamic test condition, however, no significant decrease of mobility of the lumbosacral joint by either of the casts could be observed. In both cast conditions, considerably more sagittal rotation was found during walking than with the other two exercises. CONCLUSION: In the sagittal plane, a plaster cast with or without unilateral hip immobilization can decrease motion during spinal flexion-extension. This stabilizing effect on the lumbosacral joint could not be observed during walking. PMID- 9201862 TI - Reliability of lumbar spine radiograph reading by chiropractors. AB - STUDY DESIGN: An intraobserver and interobserver study on the reproducibility of data was performed. OBJECTIVES: This study investigates the variability in the interpretation of lumbar spine radiographs by chiropractors working in private practice. SUMMARY OF BACKGROUND DATA: In chiropractic practice radiographs are used often, but this use is currently under debate. Therefore, there is a need for further study of the value of diagnoses made by radiographs by chiropractors. An acceptable intra- and interobserver agreement in radiograph reading is a prerequisite for a useful application of radiographs as a diagnostic tool in daily practice and in research. METHODS: Four chiropractors read 100 blinded sets of standard, erect anteroposterior and lateral lumbar radiographs independently. The same set was read in two separate sessions with a 2-month interval. The first session revealed the interobserver agreement. The comparison of the ratings by the same assessor in the two sessions indicated the intraobserver agreement. The assessors used a specially developed criteria list with emphasis on "nonspecific" radiographic findings. The prevalence of some important categories was increased artificially. Agreement was expressed in percentage agreement and generalized kappa, combining the results of all four assessors. RESULTS: Most kappas ranged from 0.40 to 0.75, representing fair to good agreement. In general, intraobserver agreement was better than interobserver agreement. The low kappas that were found may be explained partially by the high-agreement-low-kappa paradox as a result of a low prevalence. CONCLUSION: The kappas and percentage agreement were acceptable, although not excellent. These results will be beneficial for future research on the value of radiograph diagnosis of nonspecific findings for delivery of safe and effective chiropractic therapy. PMID- 9201863 TI - Risk indicators of low back pain among workers in Japan. Association of familial and physical factors with low back pain. AB - STUDY DESIGN: A questionnaire was given to 3,042 Japanese workers at a factory in 1992. It surveyed age, gender, weight, height, job classification, and work environment, as well as the perceived causes, onset age, and characteristics of low back pain. Family history of low back pain among first-degree relatives and perception of physical condition also were assessed. OBJECTIVES: To investigate the risk indicators of low back pain in Japanese workers with various kinds of job classifications in a manufacturing company. SUMMARY OF BACKGROUND DATA: Risk indicators of low back pain in Japanese patients have not been fully investigated in previous studies. METHODS: Prevalence rates and characteristics of low back pain were examined among 3,042 factory workers (2,517 men and 525 woman) with jobs with varying physical demands. In the analysis of risk indicators of low back pain, the odds ratios and 95% confidence intervals were computed. In addition, a multiple logistic analysis was performed to evaluate risk indicators of low back pain. RESULTS: Point and lifetime prevalence of low back pain were correlated with the physical demands of the job. The perceived cause found to be most associated with low back pain were lifting in workers with jobs requiring moderate to heavy work and sports activity in sedentary workers. Family history of low back pain in parents, siblings, and children was a risk indicator of low back pain. The average age of the first attack of low back pain in workers with a family history of it in their parents was significantly younger than that in workers with no family history in a multiple logistic analysis in male workers, the physical work demands, age, and family history of low back pain in their parents were risk indicators; however, obesity was not a risk indicator. Physical and mental conditions of workers also were associated with low back pain. CONCLUSIONS: The physical job demands show a clear association with the point and lifetime prevalence of low back pain, and improvements in work conditions may decrease low back symptoms among workers. It is likely that a family history of low back pain and physical and mental conditions of workers also should be considered in the management of low back pain. PMID- 9201864 TI - Reliability of roentgenogram evaluation of pedicle screw position. AB - STUDY DESIGN: This was a human cadaver study of the accuracy of biplanar roentgenography in determining pedicle screw position. OBJECTIVE: To determine the independent accuracy of radiologic evaluation of screw placement and to determine if there are any particular screw malpositions that are more likely to produce a false sense of acceptable screw position. SUMMARY OF BACKGROUND DATA: Other investigators have reported the correlation between radiologic evaluation and anatomic dissection. However, in those studies the radiologic evaluation was not independent of the surgeons placing the screws. There has been no comment in the literature regarding particular screw malpositions that would lead the surgeon into a false sense of successful screw placement. METHODS: Pedicle screws were placed in cadaver spines, and biplanar roentgenograms of the specimens were evaluated by independent observers. The results of the roantgenogram evaluation then were compared to those of the anatomic dissection. RESULTS: The accuracy of roentgenogram evaluation varied from 73% to 83%, depending on the experience of the surgeon grading the roentgenograms. Screws misplaced medially into the spinal canal are more likely to give the surgeon a false sense of successful screw placement. CONCLUSIONS: The surgeon must not rely solely on the roentgenograms, but instead continue to use tactile sensory skills, anatomic knowledge, and additional modalities such as electromyography monitoring. PMID- 9201865 TI - Improved accuracy of pedicle screw insertion with computer-assisted surgery. A prospective clinical trial of 30 patients. AB - STUDY DESIGN: A prospective clinical trial was done to study the accuracy of pedicle screw placement in 30 consecutive computer-assisted orthopedic surgeries. OBJECTIVES: To determine the accuracy and clinical applicability of this new method for pedicle screw insertion. SUMMARY OF BACKGROUND DATA: Conventional screw insertion techniques have been associated with high pedicle screw malplacement rates in cadaver studies and clinical studies with postoperative computed tomography evaluation. METHODS: Thirty transpedicular, low-back, titanium instrumentations were performed with computer-assisted orthopedic surgery. The accuracy of screw placement was evaluated using a sophisticated computed tomography protocol. RESULTS: The total number of pedicle screws was 174. Of these, 139 (79.9%) could be inserted with computer-assisted orthopedic surgery. The malplacement rate of computer-assisted orthopedic surgery screws was 4.3%. In screws that were not inserted by computer-assisted orthopedic surgery, the malplacement rate was 14.3%. One malplaced screw that had not been inserted with computer-assisted orthopedic surgery caused L4 root paresis. CONCLUSIONS: The accuracy of pedicle screw placement using computer-assisted surgery proved to be superior to the accuracy obtained when using conventional techniques. PMID- 9201866 TI - Endoscopic transiliac approach to L5-S1 disc and foramen. A cadaver study. AB - STUDY DESIGN: The toros of fresh cadavers were used to create endoscopic channels through the iliac wings to gain access to the L5-S1 disc and foramen. The spine and pelvis then were dissected out en bloc, and the anatomic relationships were studied. OBJECTIVES: To determine the feasibility of a transiliac approach to the L5-S1 disc and foramen and to assess the safety of this approach by studying the anatomic relationships of the transiliac track. SUMMARY OF BACKGROUND DATA: Because of its location deep in the pelvis, the L5-S1 disc and foramen are not easily accessible via a supra-iliac portal. A laparoscopic approach violates the abdominal cavity, and makes major vessels and viscera at risk for injury. METHODS: A core drill was inserted over a guide wire into the iliac wing under fluoroscopy to obtain a core of bone, which then was removed to create a transiliac channel. An arthroscope was inserted through the channel to perform discoscopy or foraminoscopy. The spine was dissected out en bloc to study the relationships of the track. RESULTS: It was possible to use a transiliac approach to L5-S1 in all the experiments. There was no damage of neural structures in any of the experiments. CONCLUSIONS: The results of this study suggest that it is possible to access the L5-S1 disc and foramen through the ilium without injuring important structures. It would be necessary to conduct a study based on an animal model and to evaluate the results before using this procedure in a clinical situation. PMID- 9201867 TI - Posterior lumbar epidural fat as a functional structure? Histologic specificities. AB - STUDY DESIGN: A topographic and histologic study was done to describe the location of the lumbar epidural fat and to find potential tissular specificities. OBJECTIVES: To search for possible histologic characteristics of posterior lumbar epidural fat, which so far has been described as semifluid tissue, and to determine whether posterior lumbar epidural fat is not a simple incidental tissue. SUMMARY OF BACKGROUND DATA: The lumbar epidural fat on two fetuses was studied. In adults, subcutaneous fat and posterior lumbar epidural fat were taken from seven corpses. The authors obtained 13 posterior lumbar epidural fat pads (two at L1-L2, three at L2-L3, six at L3-L4, and two at L4-L5) and four subcutaneous fat pads. METHODS: The authors studied abdominal axial histologic sections in two fetuses, histologic multiplanar sections in seven adults, and semithin sections in four adults of posterior lumbar epidural fat and subcutaneous fat. RESULTS: Fetal distribution of epidural fat was circumferential. Adult epidural fat distribution was limited to the posterior part of the vertebral canal and located at the disc level. Fascicles of connective tissue were less numerous and thinner in posterior lumbar epidural fat than in subcutaneous fat. Organized sliding spaces were found in the posterior epidural fat ped. CONCLUSIONS: Posterior lumbar epidural fat is not a simple incidental tissue and shows specific histologic features: sliding spaces and rarefaction of connective tissue that could explain semifluid features of the tissue. These characteristics suggest a functional role of posterior epidural fat in the lumbar spinal unit. PMID- 9201868 TI - Spine update. Administrative databases in spine research. AB - The use of administrative health care databases for the storage and retrieval of information is increasing. The data collection, entry, and collation follows a predictable process for hospital admissions. Many conclusions have been drawn from research performed using administrative databases. These conclusions can have significant and important implications for patients, providers, and society at large, to the extent that such data inform participants in the current health care policy debate. In an effort to better understand the significance of conclusions drawn from studies that rely on electronic administrative databases as their source of information, the present report addresses the process, strengths, weaknesses, and future plans for the use of administrative databases in spine research. PMID- 9201869 TI - Traumatic spondylopelvic dissociation: a case report and literature review. PMID- 9201870 TI - Color Doppler imaging and ovarian tumor angiogenesis: the Janus approach. PMID- 9201871 TI - Deciphering the hieroglyphics of venous Doppler velocities. PMID- 9201872 TI - Computerized analysis of behavior in fetuses with congenital abnormalities. AB - An observational study was undertaken to evaluate a computerized fetal behavior program in a clinical setting. Behavior of normal fetuses was compared with that of fetuses with a variety of congenital abnormalities. Forty-three fetuses were studied at 28-36 weeks; 26 were normal (49 recordings) and 16 had congenital abnormalities (26 recordings; ten had structural abnormalities of the central nervous system, one had Down's syndrome and five had other abnormalities). Behavior was recorded with the use of Doppler ultrasound. The duration of each recording was 60 min or more in all but two instances. The behavioral criteria studied were (1) the relative percentage of time spent in low and high fetal heart rate (FHR) variation; and (2) the percentage of time fetal activity was detected in low and high FHR variation. Fetuses with abnormalities exhibited varying patterns of behavior: only eight had patterns of FHR variation that were within the 10th and 90th centiles for normal fetuses and only one fetus exhibited a fetal activity pattern that was between the 10th and 90th centiles for the normal group. All the abnormal fetuses had a FHR pattern and/or fetal activity rate outside the 10th or 90th centiles. This study suggests that such a computerized behavioral analysis program may serve as a functional adjunct to the evaluation of a fetus with structural abnormalities. PMID- 9201873 TI - Lethal congenital arthrogryposis presents with increased nuchal translucency at 10-14 weeks of gestation. AB - This study examines the ultrasonographic features of congenital lethal arthrogryposis. In 27 cases of arthrogryposis diagnosed in the second and third trimesters there was severe bilateral talipes, fixed flexion deformities of the wrists and elbows and either fixed flexion or extension of the knees. In seven (26%) of the cases there was nuchal edema. In two fetuses with arthrogryposis that were examined at 13 weeks of gestation the nuchal translucency thickness was above the 99th centile of the normal range for crown-rump length. In three other women with previously affected pregnancies, ultrasound examination at 10-14 weeks demonstrated normal fetal nuchal translucency thickness and none of these fetuses were subsequently found to have arthrogryposis. These findings suggest that lethal arthrogryposis, which is usually diagnosed by the demonstration of multiple joint contractures during the second or third trimester of pregnancy, may present as increased nuchal translucency thickness at 10-14 weeks of gestation. PMID- 9201874 TI - Early sonographic diagnosis of Jarcho-Levin syndrome: a prospective screening program in one family. AB - The purpose of this study was to evaluate the possibility of early diagnosis of Jarcho-Levin syndrome by ultrasound examination of the fetus. Over a period of 5 years, nine women from one Arab family, known to carry an autosomal recessive form of the disease, were prospectively and repeatedly examined using ultrasound. Out of eight pregnancies, four fetuses were diagnosed as being affected by the disease as early as 12 gestational weeks. Three elected to terminate the pregnancy before viability and one was born at term. There were no misdiagnoses. We conclude that early prenatal ultrasonographic diagnosis of Jarcho-Levin syndrome is feasible, although later sonographic confirmation is often warranted. PMID- 9201875 TI - In vitro demonstration of inhibition of retrograde flow in the human umbilical vein. AB - This study was designed to investigate the apparent inhibition of retrograde flow in umbilical veins. While flushing fluid through cord veins for another study it appeared that there existed an impedance to retrograde flow. Simple hydraulic flow and Doppler ultrasound were used to evaluate this observation. First, in vitro studies were carried out on 100 normal umbilical cords to measure the time it took fluid to flow forward through a 30-cm segment of umbilical vein. These data were then compared to the time required for flow in the opposite direction. The average time was 8.84 s in the antegrade direction and 9.97 s in the retrograde direction. This difference was statistically significant with a p value of < 0.001. To evaluate this observation under more controlled conditions the second phase of the study was conducted. During phase two, umbilical cords were collected from term uncomplicated pregnancies. Doppler test fluid was pumped through the cord at a rate based on newborn weight. The volume and velocity were measured in both directions by Doppler ultrasound. Average flow volume was 369.1 ml/min in the antegrade direction and 174.8 ml/min in the retrograde direction. The velocities were 9.1 cm/s and 5.5 cm/s, respectively. The difference in mean flow volume was 194.3 ml/min. The difference of the mean velocity was 3.58 cm/s. Both are statistically significant with p-values of < 0.01. These data suggest there is an intrinsic inhibition of retrograde flow in the umbilical vein. The etiology and significance of this finding are discussed. PMID- 9201876 TI - Reversal of diastolic flow in the middle cerebral artery of the fetus during the second half of pregnancy. AB - This study obtained data on 22 fetuses in whom reversal of diastolic flow in the middle cerebral artery (MCA) was seen. In 59% of cases there was normal heart function and in 28% there was isolated tricuspid valve regurgitation. The majority (73%) of cases presented with normal fetal anatomy and most (82%) had a normal amniotic fluid index and normal fetal growth. In all cases, Doppler results of the umbilical artery and vein were normal. Of the women, 65% were on no medication and 73% did not smoke during pregnancy. The reversal of diastolic flow in the MCA was seen temporarily in the majority of cases (for 2-30 min); however, in one case with rhesus factor disease and a rim of ascites, it was observed for a longer period (about 2 h) and on the following day, intrauterine demise was recorded. In most of the cases without structural malformations, the neonatal outcome was normal. We conclude that reversed diastolic flow in the MCA is a rare and usually transient event. In one of our cases prolonged reversed flow preceded intrauterine demise. Therefore, this may be an ominous sign and careful fetal surveillance should be undertaken when this observation is made. There are a few possible pathomechanisms of reversed diastolic flow in the MCA. In the majority of cases the cause of the observed phenomenon remains unknown, but an increased pressure in the right ventricle and possible tricuspid regurgitation should be considered. PMID- 9201877 TI - Doppler assessment of the uterine and uteroplacental circulation in the second trimester in pregnancies at high risk for pre-eclampsia and/or intrauterine growth retardation: comparison and correlation between different Doppler parameters. AB - During a 20-month period we studied 175 pregnant women at high risk for hypertensive disorders of pregnancy or intrauterine growth retardation, and 172 patients at low risk, in a prospectively designed cross-sectional trial. Using duplex pulsed wave Doppler ultrasound, we recorded blood velocity waveforms from both main uterine arteries, the uteroplacental arteries in the region of placental implantation and the umbilical artery at 21-24 weeks of gestation. Persistent notches in the main stem uterine arteries and elevated resistance indices of > 0.68 in the uterine arteries and > 0.38 in the uteroplacental arteries were defined as abnormal waveforms. The incidence of proteinuric pregnancy-induced hypertension (PPIH) and intrauterine growth retardation (IUGR) were recorded as main outcome measures. Doppler proved to be more efficient at predicting a complicated pregnancy in those patients who were at high risk: a positive medical history alone was associated with a three-fold greater risk of developing PPIH and/or IUGR. In the high-risk group a single pathological Doppler sign accounted for an additional three- to four-fold increased risk, and the combination of all three pathological signs, a seven-fold additional risk for later disease. In this group PPIH and/or IUGR was found in 58.3%, compared to 8.3% if Doppler results were normal. The criterion for the definition of pathological Doppler results, whether persistent notching, the resistance index (RI) of the main stem uterine artery, or the RI in the arteries of the uteroplacental bed, was of minor importance, as all Doppler parameters were strongly correlated. However, the combination of all parameters was superior to a single parameter, and a bilateral notch was superior to a unilateral notch in terms of minimizing false-positive results. However, Doppler was less powerful in the population at low risk. Here PPIH and/or IUGR was seen in 6.1-6.4% in the group with abnormal Doppler findings compared to 5.2% in pregnancies with normal findings. None of the patients showed bilateral notching. In conclusion, pathological Doppler velocimetry of the uterine and uteroplacental circulation was a powerful predictor of PPIH and/or IUGR in high-risk pregnancies, identifying a group in which 58.3% would suffer from disease later in pregnancy. A combination of several Doppler parameters was superior to a single parameter, although the parameters were strongly correlated with each other. PMID- 9201878 TI - Transvaginal ultrasound and computed tomography combined with clinical parameters and CA-125 determinations in the differential diagnosis of persistent ovarian cysts in premenopausal women. AB - The purpose of this prospective study was to compare the accuracy of computed tomography (CT) and transvaginal ultrasonography in the differential diagnosis of persistent cystic ovarian lesions. The candidates for this study were 161 premenopausal non-pregnant women with an adnexal mass. After a 3-month follow-up, 83 masses persisted and were examined by both techniques before surgery. We also evaluated the CA-125 plasma levels. The CT and ultrasonographic diagnoses were then compared with the histopathological diagnosis. The overall agreement between the test results and the actual outcome was calculated by means of the kappa statistic. Transvaginal ultrasonography has a closer accuracy in the diagnosis of serous cysts and serous cystadenoma, ovarian carcinoma and endometrioma (value of kappa: 0.78, 0.73 and 0.80, respectively) than CT, even if the latter is associated with clinical and biochemical parameters such as patient's age and CA 125 plasma levels. Only in the diagnosis of cystic teratoma, is transvaginal ultrasonography less accurate than CT. In conclusion, in premenopausal women, transvaginal ultrasonography remains a cost-effective method in the diagnosis of most cystic ovarian lesions. PMID- 9201880 TI - Abnormal pulmonary venous return diagnosed prenatally by pulsed Doppler flow imaging. AB - Pulsed Doppler ultrasound has been used to characterize distinctive pulmonary venous flow patterns in the normal fetus and child. Changes in these characteristic flow patterns have been related to abnormal atrial and ventricular hemodynamics. We report a case of total anomalous pulmonary venous return diagnosed prenatally because of an abnormal pulsed Doppler echocardiographic flow pattern, even though color flow mapping appeared to demonstrate normal pulmonary venous drainage. This case demonstrates the importance of obtaining pulsed Doppler pulmonary venous flow profiles during fetal echocardiography, especially in cases of complex congenital heart disease. PMID- 9201879 TI - Doppler findings in chronic ectopic pregnancy: case report. AB - Chronic ectopic pregnancy is an uncommon form of tubal pregnancy manifested as a pelvic mass with minimal symptoms and a low or absent titer of human chorionic gonadotropin. For this reason, most of the reported cases have been diagnosed only after explorative laparotomy. The value of Doppler ultrasonography for preoperative diagnosis of this entity has not yet been established. We report on a 36-year-old patient who was admitted for intermittent right lower quadrant abdominal pain of 3 months' duration, and a right adnexal mass found on pelvic examination. On Doppler ultrasonography, a right complex adnexal mass was demonstrated, characterized by extensive external vascularization, aberrant vessels and arteriovenous shunting, but with no internal blood flow. Explorative laparotomy revealed a right tubal mass adherent to the omentum, and covered by numerous enlarged and tortuous blood vessels originating in the omentum. Pathological examination of the mass revealed a chronic ectopic pregnancy. The possible contribution of Doppler-specific characteristics for the diagnosis of chronic ectopic pregnancy is described and discussed. PMID- 9201881 TI - Intra-amniotic debris identified at ultrasound scanning: a feature of congenital ichthyosis. AB - A case is described in which intra-amniotic debris was identified during an ultrasound scan in a woman with polyhydramnios at 33 weeks' gestation. At delivery the neonate was found to have severe ichthyosis involving the whole of her head, trunk and limbs, with large plaques of hyperkeratotic skin on the palms of her hands and the soles of her feet. The finding of excessive debris in association with polyhydramnios should raise the possibility of an exfoliative skin disorder. PMID- 9201882 TI - Diagnosis of occult colon cancer: possible new use of transvaginal color imaging. PMID- 9201884 TI - False-positive diagnosis of fetal face malformation due to brow compression in early labor. PMID- 9201883 TI - An unusual late presentation of hematocolpos diagnosed by sonography. PMID- 9201886 TI - [72nd annual meeting of the Japanese Society for Tuberculosis. Sapporo City. June 12-13, 1997. Abstracts]. PMID- 9201887 TI - [Regional meeting of the Japanese Society of Otolaryngology. 1996. Abstracts]. PMID- 9201885 TI - Report from the first Kenneth Gottesfeld-Charles Hohler Memorial Foundation conference on obstetric and gynecological ultrasound. PMID- 9201888 TI - [39th annual meeting of the Japan Geriatrics Society. Tokyo. June 18-20, 1997. Abstracts]. PMID- 9201889 TI - [Vaccination, immunobiological agents safety and citizen's rights]. PMID- 9201890 TI - Studies on mosquitoes (Diptera:Culicidae) and anthropic environment. 12-host seeking behaviour of Anopheles albitarsis s.l. in south-eastern Brazil. AB - Results obtained with Anopheles albitarsis s.l. catches mainly performed through human bait at the Ribeira Valley region, SP (Brazil), are reported. Two species of the complex were recognized, namely An. albitarsis s.s. and species B. This latter predominated both in the rice fields and in the dwelling environments. The crepuscular rhythms showed an unimodal sunset pattern with most blood-seeking females caught during dusk. The absence of differences between indoor and outdoor behavior was confirmed for both species of the complex. PMID- 9201891 TI - Mortality among patients with non-affective functional psychoses in a metropolitan area of south-eastern Brazil. AB - High mortality rates among those suffering from schizophrenia and related psychoses have been consistently described in developed societies. However, to date there is a lack of data on this matter in Brazil. In order to examine this issue, a prospective 2-year follow-up study was carried out in S. Paulo. The sample consisted of 120 consecutive admissions to psychiatric hospitals in a defined catchment area, aged 18 to 44 years old, with clinical diagnoses of non affective functional psychoses according to the ICD-9. After 2 years, 116 (96.7%) subjects were traced. During the study period there were 7 deaths (6.0% of those traced), 5 (4.3%) due to suicide. All but one of the suicides occurred in the first year after discharge from hospital. Age and sex Standardised Mortality Ratios (relative to rates for the population of the city of Sao Paulo) were 8.4 for overall mortality (95% confidence interval: 4.0-15.9) and 317.9 for deaths due to suicide (95% confidence interval: 125.2-668.3). These results are in agreement with previous studies, and show that in Brazil non-affective functional psychoses are life-threatening illnesses, which need adequate care, particularly when patients go back to live in the community after hospital discharge. PMID- 9201892 TI - ["Avoidable" infant mortality in two cities of northeastern Brazil: quality indicator of the local health system]. AB - This paper seeks to discover the magnitude and causality structure of infant mortality--considered a "sentinel even" for quality-of-care indexes in health--in two municipalities of Northeastern Brazil. This is a population based study of the "invoked experimentation" type comparing observed infant mortality with that expected, given a properly functioning maternal and infant care program, allowing for the calculation of a "preventable index" (PDI) for these two municipalities. The preliminary step consisted of an active search and epidemiological investigation of deaths in order to eliminate their underreporting as events. Infant mortality rates in the two areas were relatively low--39 and 44 per thousand live births, respectively--but PDI in both was classified of the order of 40%, thus indicating a causality structure compatible with mortality rates of 100 per thousand. These findings suggest an uneven distribution of deaths, proved by an analytical comparison of the low income population with that of other income brackets (with risk ratios of 8 and 17.6 for total infant mortality and infant mortality from infectious diseases, respectively). PDI proved to be a useful index of preventability of infant deaths, and has the advantage of being simple and easy for health system managers concerned with the quality of health programs devoted to mothers and children to use. PMID- 9201893 TI - [Youth mortality: analysis of the period from 1930 to 1991 (the epidemiological transition to violence)]. AB - Youth (15 to 24 years old) mortality in the cities of Rio de Janeiro and S. Paulo from 1930 to 1991 is studied. The objective is the recovery of historical data covering the period from the third decade of this century up to the present so as to evaluate mortality profile changes based on causes of death and to compare them with international indicators. Results show that S. Paulo experienced a rapid decline in the death rate for the group up to 1970, as also happened in Rio de Janeiro city up to 1980. This latter city has always shown higher mortality rates. However, during the past decade a higher proportion of deaths occurred in S. Paulo resulting in closer mortality curves. Young people's mortality rates are not no longer decreasing. The rising tendency is accounted on male mortality increase. Infectious diseases, primarily tuberculosis, were responsible for the highest rates during the first decades studied, up to the fifties. After 1960 a transition took place and violent deaths, such as accidents and homicide, became the leading causes. Besides them, depending on the period analysed, cardiovascular diseases, respiratory infection and, later on, AIDS came to occupy a prominent position. PMID- 9201894 TI - [Behavior of hemolymph amebocytes from planorbidae in the presence of Schistosoma mansoni larvae parasitism, after inoculation of Indian ink or fracture of the shell]. AB - The behavior of the hemolymph amebocytes of Biomphalaria glabrata and Biomphalaria tenagophila was studied by means of the infection by the Schistosoma mansoni, using inoculation with Indian ink and fracture of the shell. A differential count of the amebocytes in the hemolymph was made with special attention given to the types of cell found in each sample. Histopathological evaluation of the molluscs exposed to the Schistosoma mansoni miracidia and analyses of the amebocytes were made. Correlation between the variance of the number of hemolymph amebocytes and tissue reaction was also made. The morphological study of amebocytes was carried out with the help of a phase microscope. An increase in granulocytes in Biomphalaria tenagophila occurred only when the molluscs were infected by Schistosoma mansoni, which suggests the specificity of the reaction to the parasitism. Comparing the results obtained with B. glabrata and B. tenagophila it is concluded that these molluscs show different amebocitary behavior in the presence of the diverse stimuli used. PMID- 9201895 TI - [Distribution of Biomphalaria straminae in the southern neotropical region of Brazil]. AB - A careful anatomical revision of 10,616 preserved specimens of snails from 76 localities of the State of S. Paulo, Brazil, was made with a view better to determining the geographical distribution of Biomphalaria straminea in the Neotropical Region of Southern Brazil. The analysis has shown that previous determinations were correct. The study was then complemented with a survey of information from the literature about distribution of the species. The distribution pattern of the species has expanded greatly over the last few years, perhaps an account of the construction of new dams, and the navigation system in the upper Parana Basin. Epidemiological data have shown that B. straminea is a good host to S. mansoni. Continuous schistosomiasis control must be exercised so as to prevent the further expansion of the disease. PMID- 9201896 TI - [Fortification of fluid milk for the prevention and treatment of iron deficiency anemia in children under 4 years of age]. AB - The effectiveness of the use of chelate amino acid iron fortified fluid milk in the treatment of iron deficiency in children under four years of age was studied. The 269 children included in this trial received 1 liter/day of fluid milk fortified with 3 mg of chelate amino acid iron and were evaluated at six monthly intervals. At the beginning of the study 62.3% of the children presented anemia. After 6 months, this percentage had decreased to 41.8% and at the end of one year to 26.4%. The greatest decreases occurred in the groups comprising the subjects who were of 12 to 23 months of age and those under one year of age. Among the children who presented initial hemoglobin levels under 9.5 g/dl, 59.3% were free of anemia after one year of follow-up. Of those presenting initial hemoglobin levels between 9.5 and 10.9 g/dl, 66.7% recovered from their anemia. There was also greater hematological improvement in the children that ingested over 750 ml/day of fortified milk in those families that did not share the supply of supplement among their other members and in those families that had only one child under five years of age. These findings led to the conclusion that the fortification of fluid milk is a viable and effective method for the treatment of iron deficiency in pre-school children. PMID- 9201898 TI - [Diabetes mellitus and ischemic heart disease: case-control study]. AB - Several works have been reported diabetes as an independent risk factor for ischaemic heart disease. The use of different methodologies have been an obstacle to a comparison of these studies. The study realized has sought to test the association between diabetes and ischaemic heart disease, after adjusting for known confounders and/or modifiers of effect. The study was designed as a case control study and the period of data collection was one year (March/93 to February/94). The cases were compared with three kinds of controls in two base populations: primary and secondary. The sample was composed of 833 individuals of both genders of 30-69 years of age living in the city of S. Paulo, Brazil. Logistic regression was the statistical method used for the analysis of the data. The results showed that diabetes was not an independent risk factor for IHD. Hypertension, hypercholesterolemia, smoking and family antecedents of cardiovascular diseases were considered major risk factors for ischaemic heart disease. The interaction between diabetes and exposure levels of the others variables did not present statistical significance. Some methodological issues are presented to explain different magnitudes of effect according to the different kinds of controls. It is concluded that the presence of major risk factors in the models contributed to the disappearance of the association between diabetes and ischaemic heart disease. PMID- 9201897 TI - [Neurological disorders in workers with low levels of lead in the blood. II- Neuropsychological disorders]. AB - This is a cross-sectional study with a randomized choice of individuals aiming at studying the validity of the Brazilian biological exposure limits applied to lead level in the blood (PbB) and delta-aminolevulinic acid in the urine (ALAU), which are 60 mu/dl and 10 mg/g.creat., respectively. Thus, twenty workers, whose PbB and ALAU values have been below these limits over the past two years, were selected at random at a battery plant in the State of S. Paulo, Brazil. The workers were submitted to a variation of the WHO Neurobehavioral Core Test Battery. The results were compared with those obtained for workers of a control group also chosen at random. The lead workers showed memory, mood and motor coordination disorders. Comparing these results with those obtained from the control group, a significant difference was observed (p-value < 0.02). The results indicate that the Brazilian biological exposure limits above should be reconsidered. PMID- 9201899 TI - [Environment and health: an analysis of intra-urban differentials in the city of Sao Paulo, Brazil]. AB - A field study undertaken in the city of S. Paulo is presented as part of the project Environment and Health in Developing Countries: An Analysis of Intra Urban Differentials Using Existing Data financed by the Ministry of Foreign Affairs of the United Kingdom with academic support from the London School of Hygiene and Tropical Medicine (LSHTM). The research aim was to fill in the gaps in the information needed to establish associations between mortality, urbanization and the environment. Statistics were produced by means of existing data collected by city departments, research carried out by universities and census data. Data quality was assessed taking into consideration data coverage, accuracy, and sensibility to pinpoint deprived areas in the city of S. Paulo. Socioeconomic data were used to create a composite index for districts and subdistrict in order to form four socioeconomic zones. Mortality differentials between the zones by five broad age groups (0-4, 5-14, 15-44, 45-64 e 65+) and four ICD chapters (circulatory, respiratory, infectious and parasitic and external causes) are presented. The zoning used showed that 43.8% of S. Paulo residents live in areas under the worst socioeconomic conditions. Health data demonstrated that people living in this areas had higher rates of mortality then those living in the areas with the best conditions. Finally, excess mortality data are analyzed and suggested as a good method for allocating health resources. PMID- 9201900 TI - [Antinomies and epistemological "sutures" between the socio-biological and the collective-individual characteristics within the field of social epidemiology]. AB - The complexity of the epidemiological object has provoked discussion of the elements that have constituted it over time sometimes assuming the form of antinomies. By using as substract fundamental texts, the formulation and the proposal for a solution of the antinomies between the social-biological and the collective-individual are been analysed. The validity of the theoretical wasy ahead indicated by the epidemiological discourse for the solution of the conflict of "laws" that surround this object are been criticized. The concept of the individual in Marx and Heller is been used to contribute to the "suturing" of the three strata of reality: the universal, the particular and the singular. PMID- 9201901 TI - [First recorded occurrence of Aedes (Stegomyia) albopictus (Skuse) in the southeastern region of Brazil]. AB - The first recorded occurrence of Aedes (Stegomyia) albopictus (Skuse) in Curitiba, Parana, South of Brazil, was described. The collection was carried out by means of aspirator in human bait. PMID- 9201902 TI - Fertility, family planning, and women's health: new data from the 1995 National Survey of Family Growth. AB - OBJECTIVES: This report shows data on a wide range of topics from the 1995 National Survey of Family Growth (NSFG), including: pregnancy and birth, marriage, divorce, cohabitation, sexual intercourse, contraception, infertility, use of family planning and other medical services, and health conditions and behavior. METHODS: The data in this report are based on in-person interviews with a national sample of 10,847 women 15-44 years of age. The interviews lasted an average of 103 minutes. The response rate was 79 percent. The sample data are adjusted for nonresponse and are national estimates. RESULTS: Following large increases in the 1970's and 1980's, the proportion of teenagers who have ever had sexual intercourse decreased slightly between 1990 and 1995; condom use, both at first intercourse and currently, has increased markedly since the 1970's. These changes may have contributed to the decreases in the teen birth rate observed in the 1990's. For all women 15-44 years of age, the number whose partner was currently using the condom (at the date of interview) increased from 3.6 million in 1982 to 5.1 million in 1988 and 7.9 million in 1995. About 8 percent of women reported that their first intercourse was not voluntary. This result is consistent with an earlier national survey. About 20 percent reported that they had been forced by a man to have intercourse at some time in their lives. About 10 percent of births in 1990-95 were unwanted by the mother compared with 12 percent in 1984-88. The decrease in unwanted births was particularly large for black women. It appears that the prevalence of pelvic inflammatory disease (PID) and vaginal douching have both decreased since 1988. PMID- 9201903 TI - High-energy channeling in protein folding. AB - Recent controversy about the role of populated intermediates in protein folding emphasizes the need to better characterize other events on the folding pathway. A complication is that these involve high-energy states which are difficult to target experimentally since they do not accumulate kinetically. Here, we explore the energetics of high-energy states and map out the shape of the free-energy profile for folding of the two-state protein U1A. The analysis is based on nonlinearities in the GdnHCl dependence of the activation energy for unfolding, which we interpret in terms of structural changes of the protein-folding transition state. The result suggests that U1A folds by high-energy channeling where most of the conformational search takes place isoenergetically at transition-state level. This is manifested in a very broad and flat activation barrier, the top of which covers more than 60% of the reaction coordinate. The interpretation favors a folding mechanism where the pathway leading to the native protein is determined by the sequence's ability to stabilize productive transition states. PMID- 9201904 TI - Activation of transducin guanosine triphosphatase by two proteins of the RGS family. AB - RGS proteins (regulators of G protein signaling) constitute a newly appreciated group of negative regulators of G protein signaling. Several members of this group stimulate the guanosine triphosphatase (GTPase) activity of various G protein alpha-subunits, including the photoreceptor G protein, transducin. In photoreceptor cells transducin GTPase is known to be substantially accelerated by the coordinated action of the gamma-subunit of its effector enzyme, cGMP phosphodiesterase (PDE gamma), and another yet unidentified membrane-associated protein factor. Here we test the possibility that this factor belongs to the RGS family of GTPase stimulators. We report a detailed kinetic analysis of transducin GTPase activation by two members of the RGS family, RGS4 and G alpha interacting protein (GAIP). RGS4, being at least 5-fold more potent than GAIP, stimulates the rate of transducin GTPase by 2 orders of magnitude. Neither RGS4 nor GAIP requires PDE gamma for activating transducin. Rather, PDE gamma causes a partial reversal of transducin GTPase activation by RGS proteins. The effect of PDE gamma is based on a decreased apparent affinity of RGS for the alpha-subunit of transducin. Our observations indicate that GTPase activity of transducin can be activated by at least two distinct mechanisms, one based on the action of RGS proteins alone and another involving the cooperative action of the effector enzyme and another protein. PMID- 9201905 TI - Structural study of the relationship between the rate of membrane fusion and the ability of the fusion peptide of influenza virus to perturb bilayers. AB - The amino-terminal segment of the HA2 protein of influenza virus (fusion peptide) has been identified as an important region for membrane fusion. The wild type virus can fuse to membranes more rapidly at pH 5 than at pH 7.4. It has been demonstrated that there is a relationship between the ability of the peptide to promote the formation of inverted phases and the fusogenicity of the intact virus. In this work, we use small-angle X-ray diffraction to study the mechanism of the structural effect of the peptide, at different pHs, on lipid systems characterized by each having a different spontaneous radius of curvature. The overall results show that the action of the peptide on the polymorphism of the lipid systems investigated is strongly pH-dependent. In particular, a rapid formation of cubic phases at pH 5.0 is observed in the presence of this fusion peptide. The ability of the fusion peptide to promote cubic phases exhibits the same dependence on the pH as does the fusogenicity of the intact virus. It is proposed that the peptide promotes cubic phases at pH 5.0 by changing the kinetics of the lamellar to inverted phase transitions. PMID- 9201906 TI - Role of tryptophan-63 of the kringle 2 domain of tissue-type plasminogen activator in its thermal stability, folding, and ligand binding properties. AB - Conservative (F and Y) and radical (H and S) mutations have been engineered at a rigidly conserved aromatic residue, W63, of the isolated recombinant kringle 2 domain of tissue-type plasminogen activator (r-K2tPA), an amino acid residue predicted from the X-ray crystal structure to be important in the ligand binding properties of this isolated protein domain. The variants were expressed in Pichia pastoris cells. The binding constants of epsilon-aminocaproic acid (EACA), 7 aminoheptanoic acid (7-AHpA), and trans-(aminomethyl)cyclohexanecarboxylic acid (AMCHA) to each of these mutant polypeptides were determined by titrations of the alterations in intrinsic fluorescence of the variant kringles with the ligands. As compared to wild-type r-K2tPA, increases in the Kd (dissociation) values of approximately 15-fold and 20-200-fold were found for the W63F and W63Y mutants, respectively, toward these three ligands. Neither the W63H nor the W63S variant interacted with these same ligands. Differential scanning calorimetric analyses were also performed on each of the peptides to determine whether the alterations affected the conformational stability of wtr-K2tPA. The data demonstrated that all of these mutants were thermally destabilized, possessing temperatures of maximum heat capacity (Tm) values that were 12-20 degrees C lower than that of wtr-K2tPA. Addition of EACA resulted in increases (approximately 12 degrees C) in the Tm values of r-[W63F]-K2tPA and r-[W63Y]K2tPA, a result showing that EACA stabilized the native conformations adopted by these kringle domains. As expected from its greatly diminished binding to r-[W63H]K2tPA and r-[W63S]-K2tPA, high concentrations of EACA had little effect on the Tm of thermal denaturation of these latter mutants. 1H-NMR analysis of the two aromatic mutant kringles was employed to assess their overall comparative folding properties. The high upfield chemical shifts (-0.98 ppm) of the CH3(delta') protons of L47, a major signal of proper kringle folding, were slightly lowered to -0.83 to -0.86 ppm in the cases of all of the mutants. This is due to alterations in the W25-L47 side-chain spatial orientations, possibly the result of slight conformational alterations that affect the distance relationships of these two amino acid side chains. Assignments of nearly all of the protons of the aromatic residues in the W63F and W63Y mutants were accomplished, and few additional differences from their wild type counterpart were noted. Reactivities of the mutants against four different monoclonal antibodies directed to wtr-K2tPA revealed the possibility that some small local conformational alterations might have resulted from the residues that have replaced the W63. We conclude that W63 possesses an important direct role in the ligand binding properties of r-K2tPA. This residue also contributes significantly to the stability of the native conformation of this kringle domain and perhaps to maintenance of local conformations. PMID- 9201907 TI - Crystal structures of a mutant (betaK87T) tryptophan synthase alpha2beta2 complex with ligands bound to the active sites of the alpha- and beta-subunits reveal ligand-induced conformational changes. AB - Three-dimensional structures are reported for a mutant (betaK87T) tryptophan synthase alpha2beta2 complex with either the substrate L-serine (betaK87T-Ser) or product L-tryptophan (betaK87T-Trp) at the active site of the beta-subunit, in which both amino acids form external aldimines with the coenzyme, pyridoxal phosphate. We also present structures with L-serine bound to the beta site and either alpha-glycerol 3-phosphate (betaK87T-Ser-GP) or indole-3-propanol phosphate (betaK87T-Ser-IPP) bound to the active site of the alpha-subunit. The results further identify the substrate and product binding sites in each subunit and provide insight into conformational changes that occur upon formation of these complexes. The two structures having ligands at the active sites of both alpha- and beta-subunits reveal an important new feature, the ordering of alpha subunit loop 6 (residues 179-187). Closure of loop 6 isolates the active site of the alpha-subunit from solvent and results in interaction between alphaThr183 and the catalytic residue alphaAsp60. Other conformational differences between the wild type and these two mutant structures include a rigid-body rotation of the alpha-subunit of approximately 5 degrees relative to the beta-subunit and large movements of part of the beta-subunit (residues 93-189) toward the rest of the beta-subunit. Much smaller differences are observed in the betaK87T-Ser structure. Remarkably, binding of tryptophan to the beta active site results in conformational changes very similar to those observed in the betaK87T-Ser-GP and betaK87T-Ser-IPP structures, with exception of the disordered alpha-subunit loop 6. These large-scale changes, the closure of loop 6, and the movements of a small number of side chains in the alpha-beta interaction site provide a structural base for interpreting the allosteric properties of tryptophan synthase. PMID- 9201908 TI - Cis-autophosphorylation of juxtamembrane tyrosines in the insulin receptor kinase domain. AB - Receptor tyrosine kinases undergo ligand-induced dimerization that promotes kinase domain trans-autophosphorylation. However, the kinase domains of the insulin receptor are effectively dimerized because of the covalent alpha2beta2 holomeric structure. This fact has made it difficult to determine the molecular mechanism of intraholomeric autophosphorylation, but there is evidence for both cis- and trans-autophosphorylation in the absence and presence of insulin. Here, using the cytoplasmic kinase domain (CKD) of the human insulin receptor, we demonstrate that autophosphorylation in the juxtamembrane (JM) subdomain follows a cis-reaction pathway. JM autophosphorylation was independent of CKD concentration over the range 6 nM-3 microM and was characterized kinetically: Half-saturation (K(ATP)) was observed at 75 microM ATP [5 mM Mn(CH3CO2)2] with a maximal rate of 0.24 mol of PO4 (mol of CKD)(-1) min(-1). Pairwise substitutions of Phe for Tyr in the other two autophosphorylation subdomains, generated by site directed mutagenesis, altered the kinetics of JM autophosphorylation but did not change the pathway from a cis-reaction. Tyr(1328,1334) to Phe (in the carboxy terminal subdomain) yielded <2-fold increase in the efficiency of JM autophosphorylation, whereas Tyr(1162,1163) to Phe (in the activation loop subdomain) yielded approximately 38-fold increased efficiency of JM autophosphorylation, due predominantly to a 23-fold decreased K(ATP). These findings demonstrate basal state binding of ATP to the CKD leading to cis autophosphorylation and novel basal state regulatory interactions among the subdomains of the insulin receptor kinase. On the basis of these results and the crystal structure of the conserved catalytic core of this kinase [Hubbard, S. R., et al. (1994) Nature 372, 746], a model is proposed which reconciles the JM cis reaction and the activation loop cis-inhibition/trans-reaction with the complex kinetics of insulin receptor autophosphorylation [Kohanski, R. A. (1993) Biochemistry 32, 5766]. PMID- 9201909 TI - Site-directed mutagenesis of dendrotoxin K reveals amino acids critical for its interaction with neuronal K+ channels. AB - Dendrotoxin K (DTXK) is a 57-residue protein from mamba venom that blocks certain non-inactivating, voltage-activated K+ currents in neurones. In order to pinpoint the residues responsible for its specificity, structure-activity relations of DTX(K) were investigated by mutagenesis. A previously cloned gene encoding this toxin [Smith et al. (1993) Biochemistry 32, 5692-5697] was used to make single mutations; after expression in Escherichia coli as fusion proteins and enzymatic cleavage of the conjugates isolated from the periplasmic space, nine toxins were purified. Structural analysis of the native DTXK and representative mutants by circular dichroism showed that no significant differences were detectable in their folded structures. The biological activity of the mutants, which contained alterations of positively charged and other amino acids, was determined from their abilities to compete for the binding of 125I-labeled DTX(K) to K+ channels in synaptic plasma membranes from rat cerebral cortex. Mutants with residues substituted in the alpha-helix near the C-terminus (R52A or R53A) yielded binding parameters similar to those of wild-type and native DTX(K). In the case of the beta-turn (residues 24-28), however, altering single amino acids reduced binding to the high-affinity site of K+ channels, with the rank order of decreases being K26A >> W25A > K24A = K28A. Also, substitutions made in the 3(10)-helix (residues 3-7), a region located close to the beta-turn, produced equivalent effects (K3A > K6A). Thus, it is deduced that residues in the distorted beta-turn and neighboring 3(10)-helix of DTX(K) are critical for its interaction with neuronal K+ channels. PMID- 9201910 TI - Calcium binding and homoassociation of E-cadherin domains. AB - Cadherins are single pass transmembrane glycoproteins which mediate calcium dependent cell-cell adhesion by homophilic interactions. To reveal the molecular details of calcium binding and homoassociation, we recombinantly expressed in Escherichia coli a domain pair consisting of the first two domains of E-cadherin (ECAD12) and the single domains 1, 2, and 5. ECAD12 encompasses the most N terminal of the four putative calcium-binding pockets in the extracellular region of E-cadherin. Equilibrium dialysis experiments revealed that the single domains do not bind Ca2+, but ECAD12 was found to bind three calcium ions. ECAD12 dimerizes (Kd = 0.08 +/- 0.02 mM) in the presence of Ca2+ as we could demonstrate by analytical ultracentrifugation. Calcium binding to ECAD12 induces conformational changes which were monitored by electrophoretic mobility and by circular dichroism. By analyzing our equilibrium dialysis data with a single binding site model, we found an average Kd of 460 microM for the three bound Ca2+. Assuming a model for three binding sites, which slightly increased the quality of the fit, we obtained two identical Kds of 330 microM and a third much higher Kd of 2 mM. The entire extracellular region of E-cadherin, which was recombinantly expressed in mammalian cells, binds nine Ca2+ with a much lower average Kd of 30 microM. Therefore, we conclude that the four calcium binding pockets are not identical. Since binding to ECAD12 occurs at Ca2+ concentrations close to those in the extracellular space, we suggest that the N-terminal domain pair might be involved in calcium regulation of E-cadherin mediated cell-cell adhesion. PMID- 9201911 TI - Identification of oxidation-sensitive peptides within the cytoplasmic domain of the sarcoplasmic reticulum Ca2+-ATPase. AB - We have examined the oxidative sensitivity of the Ca2+-ATPase of skeletal muscle sarcoplasmic reticulum (SR) membranes, exposing isolated SR membranes to the thermolabile water soluble free radical initiator, 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH). Incubation with up to 702 microM AAPH-derived radicals results in a concentration- and time-dependent inhibition of calcium-dependent ATPase activity correlating with the loss of monomeric Ca2+-ATPase polypeptides, and the concomitant appearance of higher molecular weight species. However, no oxidant-induced protein fragmentation is detected. The observed formation of oxidant-induced bityrosine accounts for the intermolecular Ca2+-ATPase cross links, as well as intramolecular cross-links. The oxidation of sulfhydryl groups to disulfides as another possible source of intermolecular cross-links has been ruled out after examination of SDS -PAGE performed under both reducing and non reducing conditions. Exposure of the SR membranes to AAPH-derived radical species results in a small degree of lipid peroxidation that is not correlated with enzyme inactivation, suggesting that modification of membrane-spanning peptides is not related to enzyme inactivation. Six cytoplasmic peptides have been identified that are modified by exposure to AAPH or, alternatively, to hydrogen peroxide, suggesting that these regions of the Ca2+-ATPase are generally sensitive to oxidants. These oxidized peptides were identified after separation by reversed-phase HPLC followed by N-terminal sequencing and amino acid analysis as corresponding to the following sequences of the Ca2+-ATPase: (i) Glu121 to Lys128, (ii) His190 to Lys218, (iii) Asn330 to Lys352, (iv) Gly432 to Lys436, (v) Glu551 to Arg604, and (vi) Glu657 to Arg671. The Glu551 to Arg604 peptide, located within the nucleotide binding domain, was found to participate in the formation of intermolecular bityrosine cross-links with the identical Glu551 to Arg604 peptide from a neighboring Ca2+-ATPase polypeptide chain. PMID- 9201912 TI - A model membrane approach to the epidermal permeability barrier: an X-ray diffraction study. AB - The permeability of mammalian skin is determined in large part by lamellar lipid domains packed between cells of the upper layer of the epidermis, the stratum comeum. Although these lamellae have features in common with typical biological membranes, they differ in having a lipid population composed mainly of ceramides, cholesterol, and free fatty acids. In our initial studies of the relationship between lipid composition and phase behavior in this unusual system, we used deuterium NMR [Kitson et al. (1994) Biochemistry 33, 6707-6715] to examine aqueous dispersions of nonhydroxylated bovine brain ceramide, cholesterol, and perdeuterated palmitic acid, and found complex phase behavior as a function of temperature and pH, whereas analogous dispersions in which sphingomyelin replaced ceramide resulted in spectra consistent with a fluid lamellar phase under the same conditions. To extend these observations, we examined the same dispersions at pH 5.2 by means of X-ray diffraction. The significant findings are as follows: (1) the ceramide dispersions form complex crystalline phases between room temperature and about 40 degrees C; (2) the majority of the crystalline cholesterol is not in a separate phase; and (3) the analogous sphingomyelin dispersions form a fluid lamellar phase under the same conditions. We conclude that ceramides, even in the presence of considerable mole fractions of cholesterol, can form crystalline lamellar structures. We suggest that the existence of such structures in stratum corneum may be important in the function of the epidermal permeability barrier, and that the interaction between ceramide and cholesterol in other biological membranes may result in regions having unique physical properties. PMID- 9201913 TI - The molar ratios of alpha and beta subunits of the Na+-K+-ATPase differ in distinct subcellular membranes from rat skeletal muscle. AB - The Na+-K+-ATPase consists of alpha and beta subunits proposed to function as an alpha-beta heterodimer. Skeletal muscle is characterized by expression of alpha1, alpha2, beta1, and beta2 subunit isoforms. The relative molar proportions of each subunit or each protein isoform are not known, yet their subcellular distribution and expression in muscles of different fiber types are markedly different. In this study, the molar ratio of each pump subunit isoform was measured in purified membranes from skeletal muscle and compared with those in kidney and brain microsomes. Recombinant proteins were used as standards to quantitate each isoform by immunoblotting in combination with measurements of [3H]ouabain binding. The results indicate that in kidney microsomes, which express predominantly alpha1 and beta1 isoforms, the alpha:beta subunit molar ratio is approximately 1:1. In brain microsomes, the sum of all alpha (alpha1, alpha2, and alpha3) and all beta (beta1 and beta2) subunits also yielded a molar ratio of approximately 1:1. In contrast, in red (oxidative) skeletal muscles, the all alpha:beta subunit ratio was 0.2 in plasma membranes and 0.4 in intracellular membranes. The ratio of alpha2 subunits to alpha1 subunits ranged from 1.6 in surface membranes to up to 7 in internal membranes, while the beta1 subunits exceeded the beta2 subunits by approximately 4-fold in all membrane fractions. Thus, intracellular membranes of red skeletal muscles contain primarily alpha2 and beta1 subunits. When these intracellular membranes were further subfractionated by velocity gradient centrifugation, the alpha2:beta1 subunit ratio was 0.5 in the faster migrating (larger) membranes and 1.0 in the slower migrating (smaller) ones. This was due to a progressive decrease in abundance of the beta1 subunits without a change in the concentration of alpha2 subunits per unit protein. The Na+-K+-ATPase hydrolytic activity was higher in the larger vesicles than in the smaller ones along the sucrose gradient. These results suggest that the ratio of beta to alpha subunits may serve to regulate the catalytic activity of the Na+-K+-ATPase in skeletal muscle. PMID- 9201914 TI - Activation of regulated actin by SH1-modified myosin subfragment 1. AB - The reactive SH1 (Cys-707) group of the myosin subfragment 1 (S1) has been used frequently as an attachment site for fluorescent and spin probes in solution and muscle fiber experiments. In this study we examined (i) the motor function of SH1 spin-labeled heavy meromyosin (HMM) in the in vitro motility assays and (ii) the effect of SH1-modified S1 on the motility of regulated actin, i.e., actin complexed with tropomyosin and troponin. N-ethylmaleimide (NEM), N-(1-oxyl 2,2,6,6-tetramethyl-4-piperidinyl)-iodacetamide (IASL), N [[(iodoacetyl)amino]ethyl]1-sulfo-5-naphthylamine (IAEDANS), and iodoacetamide (IAA) were used to selectively modify the SH1 group on S1; the SH1 group on HMM was labeled with IASL. In the in vitro motility assays, 10-20% of unregulated actin filaments moved at a speed of approximately 1 microm/s over a surface coated with 90-95% modified IASL-HMM. Actin sliding was not observed with 95-98% modified IASL-HMM. The sliding of regulated actin over unmodified HMM was activated by the addition of S1 modified with any of the SH1 reagents to the in vitro motility assay solutions; both the speeds and the percentage of the moving filaments increased at pCa 5, 7, and 8. To shed light on the activation of regulated actin sliding by SH1-modifed S1, acto-S1 ATPase and the binding to actin were determined for IASL-S1. While the binding affinities to actin were similar for IASL-S1 and unmodified S1 in the presence and absence of ADP and ATP, the Km and Vmax values were approximately 10-fold lower for the modified protein. It is concluded that the activation of regulated actin by SH1-modifed S1 facilitates the interaction of unmodified HMM heads with actin and thus can increase the sliding speeds and the percentage of regulated actin filaments that move in the in vitro motility assays. PMID- 9201915 TI - The nucleotide binding folds of the cystic fibrosis transmembrane conductance regulator are extracellularly accessible. AB - Analysis of the primary sequence of the cystic fibrosis transmembrane conductance regulator (CFTR) has suggested the presence of two predicted cytoplasmic regions of the protein which are thought to be nucleotide binding folds (NBF1 and NBF2). Previous studies have shown that purified recombinant NBF1 can form anion conducting channels in planar phospholipid bilayers [Arispe et al. (1992) Proc. Natl. Acad. Sci. U.S.A. 89, 1539-1543] and that the bacterial His P protein (analogous to a NBF) can be extracellularly labeled with a membrane-impermeant reagent [Baichwal et al. (1993) Proc. Natl. Acad. Sci. U.S.A. 90, 620-624]. Based on these observations, it is reasonable to hypothesize that the NBFs from the CFTR are associated with the plasma membrane and have extracellularly-accessible regions. Direct biochemical evidence for this was obtained by determining the ability of the individual NBFs, expressed in intact Hi5 cells, to be chemically modified with the membrane-impermeant reagent NHS-biotin. The results indicate that both NBF1 and NBF2, in intact cells, can be chemically modified by extracellular NHS-biotin. The negative control, the cytosolic enzyme beta galactosidase, was not significantly labeled under these conditions, verifying the extracellular nature of the labeling reaction. When the surface-accessibility of a NBF1 construct containing the CF-causing mutation deltaF508 was analyzed, similar labeling was observed, suggesting that the mutation does not affect this aspect of the CFTR's structure. These data support the conclusion that, under certain conditions, the NBFs are capable of spanning the plasma membrane, perhaps constituting a portion of the CFTR's ion conducting channel. PMID- 9201916 TI - Structural and functional consequences of substitutions at the tyrosine 55-lysine 104 hydrogen bond in Escherichia coli inorganic pyrophosphatase. AB - Tyrosine 55 and lysine 104 are evolutionarily conserved residues that form a hydrogen bond in the active site of Escherichia coli inorganic pyrophosphatase (E PPase). Here we used site-directed mutagenesis to examine their roles in structure stabilization and catalysis. Though these residues are not part of the subunit interface, Y55F and K104R (but not K104I) substitutions markedly destabilize the hexameric structure, allowing dissociation into active trimers on dilution. A K104I variant is nearly inactive while Y55F and K104R variants exhibit appreciable activity and require greater concentrations of Mg2+ and higher pH for maximal activity. The effects on activity are explained by (a) increased pK(a)s for the catalytically essential base and acid at the active site, (b) decreases in the rate constant for substrate (dimagnesium pyrophosphate) binding to enzyme-Mg2 complex vs enzyme-Mg3 complex, and (c) parallel decreases in the catalytic constant for the resulting enzyme-Mg2 substrate and enzyme-Mg3-substrate complexes. The results are consistent with the major structural roles of Tyr55 and Lys104 in the active site. The microscopic rate constant for PPi hydrolysis on either the Y55F or K104R variants increases, by a factor of 3-4 in the pH range 7.2-8.0, supporting the hypothesis that this reaction step depends on an essential base within the enzyme active site. PMID- 9201917 TI - Crystal structure of holo inorganic pyrophosphatase from Escherichia coli at 1.9 A resolution. Mechanism of hydrolysis. AB - Crystalline holo inorganic pyrophosphatase from Escherichia coli was grown in the presence of 250 mM MgCl2. The crystal structure has been solved by Patterson search techniques and refined to an R-factor of 17.6% at 1.9 A resolution. The upper estimate of the root-mean-square error in atomic positions is 0.26 A. These crystals belong to space group P3(2)21 with unit cell dimensions a = b = 110.27 A and c = 78.17 A. The asymmetric unit contains a trimer of subunits, i.e., half of the hexameric molecule. In the central cavity of the enzyme molecule, three Mg2+ ions, each shared by two subunits of the hexamer, are found. In the active sites of two crystallographically independent subunits, two Mg2+ ions are bound. The second active site Mg2+ ion is missing in the third subunit. A mechanism of catalysis is proposed whereby a water molecule activated by a Mg2+ ion and Tyr 55 play essential roles. PMID- 9201918 TI - Mechanism-based inhibitor discrimination in the acyl-CoA dehydrogenases. AB - The catalytically essential glutamate base in the acyl-CoA dehydrogenase family is found either on the loop between J and K helices (e.g., in short-chain, medium chain, and glutaryl-CoA dehydrogenases) or on the G helix (long-chain and isovaleryl-CoA dehydrogenases). While active-site bases at either position are functionally equivalent with respect to alpha-proton abstraction, reactions that require removal of a gamma-proton show marked differences between the two enzyme classes. Thus short-chain, medium-chain, and glutaryl-CoA dehydrogenase are rapidly inactivated by 2-pentynoyl-CoA with abstraction of a gamma-proton, whereas isovaleryl-CoA dehydrogenase is not significantly inhibited. This resistance is not due to weak binding: the complex between isovaleryl-CoA dehydrogenase and 2-pentynoyl-CoA shows a Kd of 1.8 microM at pH 7.6. Migration of the catalytic base to the loop between J and K helices (using the Glu254Gly/Ala375Glu double mutant) makes isovaleryl-CoA dehydrogenase sensitive to irreversible inhibition by 2-pentynoyl-CoA. Molecular modeling suggests that this mutation brings the catalytic base close enough to abstract a gamma-proton from the bound inhibitor. Experiments with two mechanism-based inactivators that target the FAD of the medium- and short-chain acyl-CoA dehydrogenases support this conclusion. 3-Methyl-3-butenoyl-CoA requires activation by alpha-proton abstraction and rapidly yields a reduced flavin adduct with wild-type isovaleryl CoA dehydrogenase. In contrast, the isomeric 3-methyl-2-butenoyl-CoA is inert toward this enzyme because it cannot be activated by gamma-proton abstraction. Molecular modeling supports these observations. This unusual selectivity toward mechanism-based inactivators provides additional discrimination between members of the acyl-CoA dehydrogenase family. PMID- 9201919 TI - Characterization of two important histidine residues in the active site of xylanase A from Streptomyces lividans, a family 10 glycanase. AB - The active site of xylanase A (XlnA) from Streptomyces lividans contains three histidine residues, two of which (H81 and H207) are almost completely conserved in family 10 glycanases. The structural analysis of the enzyme shows that H81 and H207 are part of an important hydrogen bond network in the vicinity of the two catalytic residues (E128 and E236). In order to investigate the role of these two histidine residues for the structure/function of XlnA, three mutant enzymes were produced at each position, namely, H81R/S/Y and H207E/K/R. The specific activity of these mutant enzymes is reduced by more than 95%, revealing the importance of these two residues for the catalytic function of XlnA. The kinetic parameters of the three more active enzymes were determined, of which mutation H207K increased the K(M) 3-fold. The k(cat) of the mutant enzymes is reduced proportionally to the specific activity. Furthermore, the pKa values of the two catalytic residues are decreased in all six mutations, demonstrating a role for H81 and H207 in the hydrogen bond network responsible for maintaining the ionization state of the two catalytic residues. In most cases, the unfolding of mutated XlnA in guanidine hydrochloride (Gdn-HCl) showed that the concentration required to denature 50% of the XlnA decreased, thus demonstrating the importance of those two residues for the stability of the enzyme. Moreover, the m value [m = d(deltaG)/d[Gdn-HCl]] for the unfolding of XlnA in Gdn-HCl is increased for each of the six mutations, suggesting that the mutant proteins have less residual structure in the denatured state than does the wild-type enzyme. PMID- 9201920 TI - Geldanamycin, a heat shock protein 90-binding benzoquinone ansamycin, inhibits steroid-dependent translocation of the glucocorticoid receptor from the cytoplasm to the nucleus. AB - When they are translated, steroid receptors are assembled into a multiprotein complex containing hsp90, p23, an immunophilin, and often some hsp70. Some of the receptors, such as that for progesterone, have nuclear localization signals that are functional in the absence of hormone, and they move into the nucleus where they exist in the same multiprotein heterocomplex with hsp90. Other receptors, such as the glucocorticoid receptor, are localized predominantly in the cytoplasm in the absence of hormone and move into the nucleus in a hormone-dependent fashion. We have previously proposed that hsp90 and the immunophilin play a role in receptor trafficking [Pratt, W. B. (1993) J. Biol. Chem. 268, 21455-21458]. In this work, we show that treatment of L cells with geldanamycin, a benzoquinone ansamycin that binds to hsp90 and disrupts its function, impedes dexamethasone dependent trafficking of the glucocorticoid receptor from the cytoplasm to the nucleus. Because geldanamycin treatment of hormone-free cells causes a rapid loss of steroid binding activity, receptors were prebound with dexamethasone by incubating cells with hormone at 0 degrees C prior to shifting the temperature to 37 degrees C for 20 min to permit receptor transformation and translocation in the presence or absence of geldanamycin. Geldanamycin does not cause steroid to dissociate from prebound receptors, and it does not inhibit hormone-mediated receptor transformation assayed by conversion to the DNA-binding state. However, as reported previously for the progesterone receptor, geldanamycin blocks assembly of the glucocorticoid receptor-hsp90 heterocomplex at an intermediate state of assembly where the receptor is bound to hsp70 and p60, both of which are required components in the assembly mechanism. Our observations support the proposal that dynamic association of receptors with hsp90 is required for receptor translocation from the cytoplasm to the nucleus. PMID- 9201921 TI - The thiocarboxanilide nonnucleoside UC781 is a tight-binding inhibitor of HIV-1 reverse transcriptase. AB - The thiocarboxanilide nonnucleoside inhibitor (NNI) UC781 inhibited HIV-1 reverse transcriptase (RT) DNA polymerase activity at a 1:1 molar ratio of inhibitor to enzyme. Inhibition was linear uncompetitive with respect to template/primer (T/P) and mixed noncompetitive with respect to deoxynucleoside triphosphate (dNTP), typical of NNI. When the RT-T/P binary complex was incubated with UC781 and then separated from unbound inhibitor, recovery of enzyme activity was slow, with only about 60% activity recovered after 25 min. The inactivation of the RT-T/P complex was prevented by the presence of a large excess of UC84, another carboxanilide NNI that interacts with this RT mechanistic form. UC781 protected the RT-T/P-dNTP ternary complex from irreversible inactivation by a photoactivatable azido analog of nevirapine, implying that UC781 binds to the NNI pocket of this RT mechanistic form. UC781 did not photoprotect either the free enzyme or the RT-T/P binary complex; however, protein fluorescence quenching studies indicated that UC781 interacted with all RT mechanistic forms, with the order of affinity being RT-T/P dNTP ternary complex > RT-T/P binary complex > free RT. Reaction progress curve analysis showed that the binding of UC781 to RT is rapid (k(on) approximately 1.7 x 10(6) M(-1) s(-1)), but that dissociation is slow (k(off) approximately 1.6 x 10(-3) s(-1)). UC781 is therefore a rapid tight-binding inhibitor of HIV-1 RT, the first NNI to demonstrate this property. PMID- 9201922 TI - Identification of a functional imperfect estrogen-responsive element in the 5' promoter region of the human cathepsin D gene. AB - 17beta-Estradiol (E2) induces cathepsin D gene expression in MCF-7 human breast cancer cells. Previous studies have identified an Sp1-imperfect estrogen responsive element (ERE) half-site [GGGCGG(N)23ACGGG] (-199 to -165) in the promoter region which forms an Sp1-estrogen receptor (ER) complex and confers E2 responsiveness on the corresponding Sp1-ERE-chloramphenicol acetyl transferase (CAT) construct. Further analysis of downstream regions of the promoter identified a CGCCC(N)3TGACC sequence (-119 to -107) which is homologous to the adenovirus major late promoter element (MLPE) and binds the ER to form a retarded band in a gel electrophoretic mobility shift assay. The corresponding promoter CAT construct is also E2-inducible. The MLPE resembles an imperfect palindromic ERE containing imperfect (5') and perfect (3') ERE half-sites; analysis of oligonucleotides with mutations in these half-sites shows that only the perfect ERE half-site is required for binding the ER, whereas both sites are required for transactivation. In vivo exonuclease III footprinting showed that treatment with E2 also enhanced binding at the MLPE site. Identification of this second functional enhancer sequence in the 5'-promoter region of cathepsin D is consistent with the increasingly complex cell-specific regulation of hormone responsive genes. PMID- 9201923 TI - Cleavage of p220 by purified poliovirus 2A(pro) in cell-free systems: effects on translation of capped and uncapped mRNAs. AB - Poliovirus protease 2A(pro) has been obtained in soluble form as a fusion protein with maltose binding protein (MBP). Addition of MBP-2A(pro) to rabbit reticulocyte cell-free systems gives rise to efficient cleavage of the initiation factor of translation p220 (eIF-4G). Translation of capped mRNA encoding the influenza virus NP protein is severely impaired in lysates in which p220 has been proteolytically cleaved. This inhibition is dependent on the concentration of mRNA added to the lysate. Thus, increasing the concentrations of mRNA substantially overcomes the blockade of NP synthesis after p220 cleavage. Notably, translation of uncapped NP mRNA is also compromised in p220-deficient rabbit reticulocyte lysates, suggesting that p220 participates in the translation of both capped and uncapped NP mRNAs. The effects of p220 proteolysis by poliovirus 2A(pro) have also been assayed on luciferase mRNA translation. Three types of mRNAs encoding for luciferase have been examined: capped, uncapped, and mRNA bearing the poliovirus 5' leader region (leader luc mRNA). Synthesis of luciferase directed by any of these mRNAs was inhibited after cleavage of p220 in rabbit reticulocyte lysates. Interestingly, supplementation of the lysate with HeLa cell extracts stimulates leader luc mRNA translation by poliovirus 2A(pro). These results indicate that activation of translation of mRNAs bearing the poliovirus leader region promoted by this poliovirus protease requires a factor present in HeLa cell extracts. These findings agree well with recent experiments implicating p220 not only in protein synthesis directed by capped mRNAs but also in the translation of naturally uncapped mRNAs. PMID- 9201924 TI - Structure and interactions of the single-stranded DNA genome of filamentous virus fd: investigation by ultraviolet resonance raman spectroscopy. AB - The filamentous bacteriophage fd is a member of the Ff class of Inovirus, which includes phages f1 and M13. Ultraviolet resonance Raman (UVRR) spectra of fd have been obtained using excitation wavelengths of 257, 244, 238, and 229 nm. Excitation at 257 nm selectively enhances Raman markers of the packaged single stranded (ss) DNA genome, while excitation at the shorter wavelengths favors the detection of Raman signals from coat protein aromatics, particularly tryptophan (W26) and tyrosine residues (Y21 and Y24) of the viral coat subunit (pVIII). The principal findings are the following: (1) Distinctive markers of dA, dC, dG, and dT residues of the packaged genome are identified in UVRR spectra of fd excited at 257 and 244 nm, despite the low DNA mass composition (12%) of the virion. (2) Raman bands of the bases of packaged ssDNA show extraordinary resonance Raman hypochromism. Raman intensity losses as large as 80% of the parent DNA nucleotide intensities are observed. This is interpreted as evidence of extensive short range interactions involving bases of the packaged genome. (3) Conversely, Raman bands of tryptophan and tyrosine residues of the coat protein generally exhibit strong hyperchromism. Typically, Raman markers of the aromatic amino acids are about 3-fold more intense in the UVRR spectrum of fd than in spectra of the free amino acids. The very high Raman cross sections for residues Y21, Y24, and W26 are indicative of unusual hydrophobic environments in the viral assembly. (4) UVRR band shifts that accompany the transfer of fd from H2O to D2O solution indicate that bases of the packaged ssDNA are readily exchanged by the solvent. Similarly, the indole N1H group of W26 is accessible to solvent, as shown by N1H -> N1D exchange in D2O solution. (5) The UVRR markers of the packaged fd genome confirm the conclusion reached previously from off-resonance Raman studies that fd DNA nucleosides favor the C3'-endo/anti conformation, rather than the C2' endo/anti conformation that is characteristic of the lowest energy structure of DNA. We conclude that nucleoside conformations of the packaged fd genome are influenced by the specific organization of ssDNA and coat protein subunits in the native virion assembly. PMID- 9201925 TI - Fidelity of binding of the guanidinium nucleic acid (DNG) d(Tg)4-T-azido with short strand DNA oligomers (A5G3A5, GA4G3A4G, G2A3G3A3G2, G2A2G5A2G2). A kinetic and thermodynamic study. AB - Short strand DNA oligomers (A5G3A5, GA4G3A4G, G2A3G3A3G2, and G2A2G5A2G2) and the guanidinium (g) linked thymidyl nucleoside d(Tg)4-T-azido associate as triplexes. The melting temperatures, Tm, the association and dissociation kinetic and thermodynamic parameters and activation energies for the triplexes were determined by UV thermal analysis. The hypochromic shift and Tm for triplex formation increases with increase in concentration and decreases with the number of mismatches. The melting temperatures are between 35 and 55 degrees C in the range of ionic strength of 0.06-0.24 and decrease with increase in ionic strength at 100 deg/(ionic strength unit). The melting and cooling curves exhibit hysteresis behavior in the temperature range 5-95 degrees C at 0.2 deg/min thermal rate. From these curves, the rate constants and the energies of activation for association (k(on), E(on)) and dissociation (k(off), E(off)) processes were obtained. The second-order rate constants, k(on), for the triplex formation at 288 K are between 10 and 500 M(-2) s(-1). Values of k(on) increase with the decrease in the ionic strength. The first order rate constants for the dissociation, k(off), at 288 K are between 10(-6) and 40 x 10(-6) s(-1) and increase with increase in ionic strength. The energies of activation for the association and dissociation processes are in the range -22 to -9 kcal/mol and 8 to 29 kcal/mol, respectively. At 6.3 x 10(-5) M/base and at the physiological ionic strength (0.15-0.30) and below, the triplex structures formed with d(Tg)4-T azido and A5G3A5 and GA4G3A4G have well-defined Tm values. The melting curves with G2A3G3A3G2 and G2A2G5A2G2 are very shallow with small hypochromic shifts denoting negligible binding at physiological ionic strength. Therefore, with the increase in the G content (mismatched base pairs) at a certain concentration (e.g., 6.3 x 10(-5) M/base), discrimination (change in fidelity) occurs in the formation and strength of binding of d(Tg)4-T-azido to d(pAn pGm) oligomers. The standard molar enthalpies for triplex formation have in general larger negative values at low ionic strength than at high ionic strength, indicating that at lower mu values the formation of triplexes of d(Tg)4-T-azido with d(pAn pGm) are more favorable. The values of deltaH(standard)(288) calculated from the activation parameters are between -17 and -49 kcal/mol, and the values of deltaG(standard)(288) are between -7.5 and -11.8 kcal/mol for A5G3A5, GA4G3A4G, G2A3G3A3G2, and G2A2G5A2G2, respectively. There is a linear relationship in the enthalpy-entropy compensation for the triplex melting thermodynamics. PMID- 9201926 TI - Time-dependent inhibition of recA protein-catalyzed ATP hydrolysis by ATPgammaS: evidence for a rate-determining isomerization of the recA-ssDNA complex. AB - The ATP analog ATPgammaS is a competitive inhibitor of the recA protein-catalyzed ssDNA-dependent ATP hydrolysis reaction. The degree of inhibition by ATPgammaS, however, changes in a time-dependent manner and is consistent with a two step binding mechanism. In the first step, ATPgammaS binds to the recA-ssDNA complex in a rapid equilibrium step (KD = 50 microM). This initial binding step is followed by an isomerization of the recA-ssDNA-ATPgammaS complex to a new conformational state in which ATPgammaS is bound with a significantly higher affinity (overall K(D) = 0.3 microM). This isomerization is followed by the slow hydrolysis of ATPgammaS to ADP and thiophosphate (0.01 min(-1)). The first-order rate constant for the ATPgammaS-mediated isomerization step (20 min(-1)), although significantly greater than the rate of ATPgammaS hydrolysis, is identical to the steady-state rate constant for the recA protein-catalyzed ATP hydrolysis reaction. These results are consistent with a kinetic model in which an ATP-mediated isomerization of the recA-ssDNA complex represents the rate determining step on the recA protein-catalyzed ssDNA-dependent ATP hydrolysis reaction pathway. PMID- 9201927 TI - Intramolecular electron transfer between [4Fe-4S] clusters studied by proton magnetic resonance spectroscopy. AB - The rate constants for the intramolecular electron transfer between the two [4Fe 4S] clusters of a series of native and genetically engineered ferredoxins have been determined by proton magnetic resonance (1H NMR) spectroscopy. The measurement relies on the properties of the signals assigned to beta-protons of the coordinating cysteines when the protein is substoichiometrically reduced: these signals include coalesced peaks arising from the fast hopping of an extra electron between the two oxidized clusters of the protein. An upper limit of significantly less than 10(5) M(-1) s(-1) for the intermolecular and an average of the order of 5 x 10(6) s(-1) for the intramolecular electron transfer rate constants of several ferredoxins have been obtained. Owing to the edge-to-edge intercluster distance of approximately 10 A derived from the crystallographic structure of Clostridium acidurici ferredoxin, the rate constant associated with the intramolecular process is as expected for a nonadiabatic redox process, assuming a reasonable value of less than 1 eV for the reorganization energy. The latter could not be determined from the temperature dependence of the rate constant since no variation was observed over the temperature range accessible in these experiments. Structural changes introduced around and between the two [4Fe 4S] clusters in Clostridium pasteurianum ferredoxin by site-directed mutagenesis have been used to probe the potential involvement of dominant electron transfer pathways between the clusters. These changes have no major effect on the value of the intramolecular electron transfer rate constant. From this analysis, no specific amino acid side chain seems to play a central role in the process. The rate constants derived in the present work may serve as a basis for the study of enzymes containing two closely spaced [4Fe-4S] clusters such as found in these ferredoxins. PMID- 9201928 TI - Nature and electronic structure of the Ni-X dinuclear center of Desulfovibrio gigas hydrogenase. Implications for the enzymatic mechanism. AB - The recent determination of the X-ray crystal structure of Desulfovibrio gigas hydrogenase has revealed that the active site is a Ni-X dinuclear center [Volbeda, A., Charon, M. H., Piras, C., Hatchikian, E. C., Frey, M., & Fontecilla Camps, J. C. (1995) Nature 373, 580-587]. This unexpected result calls for a re examination of the magnetic and redox properties that have been attributed previously to a mononuclear Ni center. We have used a combination of dosimetric and electron paramagnetic resonance (EPR) techniques to investigate the nature and the electronic structure of the Ni-X center in the redox forms of D. gigas hydrogenase giving EPR signals. The metal atom X was first shown to be Fe by accurate metal content analyses. Next, by determining the EPR characteristics of a polycrystal powder, it was shown that the redox form of the enzyme studied in the X-ray crystal experiments was essentially Ni-A. The temperature dependence of the Ni-A, Ni-B, Ni-C, and Ni-L EPR signals was studied over a large temperature range. No deviation from Curie's law could be detected, which places strong constraints upon the magnitude of the possible magnetic interactions between the Ni and Fe centers. When these results and the other available magnetic data are analyzed in the light of the crystal structure, it is concluded that the Fe center is diamagnetic in all the redox states of the enzyme. On the basis of these results, a mechanistic scheme consistent with a large body of experimental data can be proposed for Ni-containing hydrogenases. PMID- 9201929 TI - Carotenoid-dependent oligomerization of the major chlorophyll a/b light harvesting complex of photosystem II of plants. AB - Under many environmental conditions, plants are exposed to levels of sunlight in excess of those required for photosynthesis. Then, a regulated increase in the rate of nonradiative dissipation of excess excitation energy in the thylakoid membrane correlates with the conversion of the carotenoid violaxanthin into zeaxanthin and provides protection from the damaging effects of excessive irradiation. The hypothesis that these carotenoids specifically control the oligomerization of the light harvesting complexes of photosystem II was tested by investigating the effects of violaxanthin and zeaxanthin on the behavior of the major complex, LHCIIb, on sucrose gradients; it was found that zeaxanthin stimulated the formation of LHCIIb aggregates with reduced chlorophyll fluorescence yield whereas violaxanthin caused the inhibition of such aggregation and an elevation of fluorescence. Measurements of 77 K fluorescence indicated that zeaxanthin was not exerting an additional direct quenching of chlorophyll fluorescence. These effects can explain the physiological control of the light harvesting system by the xanthophyll cycle. PMID- 9201930 TI - Specific photoaffinity labeling of Tyr-49 on the light chain in the steroid combining site of a mouse monoclonal anti-estradiol antibody using two epimeric 6alpha- and 6beta-(5-azido-2-nitrobenzoyl)amidoestradiol photoreagents. AB - A mouse monoclonal anti-7-(O-carboxymethyl)oximinoestradiol antibody was photoaffinity labeled with two cross-reactive 6alpha- and 6beta-(5-azido-2 nitrobenzoyl)amido[17alpha-3H]estradiol photoreagents (6alpha- and 6beta-ANBA [17alpha-3H]estradiol). Covalently bound radioactivity was found exclusively on the light chain. The maximal level of specific incorporation was 0.18 mol of label per mole of antibody for both photoreagents. In both cases, tryptic digestion of the photolabeled light chain, immunopurification with the immobilized antibody, reverse-phase liquid chromatography, and Edman degradation showed the presence of radioactive peptide GLM-([3H]X)-HGNTLEDGIPSR derived from peptide 46-61 of the light chain sequence (determined from cDNA) in which the unidentified amino acid corresponding to X is a Tyr residue. Two other radioactive peptides were also isolated, one corresponding probably to the methionine sulfoxide derivative of the peptide 46-61 photolabeled with the 6beta reagent and the other to the N-terminal tetrapeptide 46-49 of the peptide 46-61 photolabeled with the 6alpha-reagent. In all cases, the main peak of radioactivity was released at the fourth Edman cycle, thus suggesting that the same Tyr-49 residue on the light chain was photolabeled. This residue is contiguous to the N-terminal amino acid of the second hypervariable complementary determining region 50-56 of light chain. Covalent labeling was confirmed by mass spectrometry of photolabeled peptides which showed molecular ion values corresponding to the addition of the photoactive 6alpha- or 6beta-ANBA-estradiol nitrene derivatives to the peptide. PMID- 9201931 TI - Tissue factor cytoplasmic domain peptide is multiply phosphorylated in vitro. AB - Human tissue factor was phosphorylated when incubated with lysates of U87-MG cells or fractions from preparative isoelectric focusing of the lysates. The cytoplasmic domain peptide, isolated following chemical cleavage at cysteine 245, focused on PhastGel IEF near pH 3.4, indicating the presence of three phosphate groups. A peptide corresponding to the carboxyl-terminal cytoplasmic domain (residues 245-263) was synthesized and shown to be a protein kinase substrate when incubated with lysates of U87-MG cells and radiolabeled ATP. As found with full-length tissue factor, the TF(245-263) peptide was phosphorylated at all three serines, but a diphosphate form was also identified. TF(245-263) was phosphorylated in the absence of calcium as well as in the presence of calphostin C, indicating that phosphorylation can be independent of protein kinase C. These results reveal that tissue factor can be multiply phosphorylated in vitro, and that the synthetic TF(245-263) cytoplasmic domain peptide serves as a model substrate. PMID- 9201932 TI - Two-state thermal unfolding of a long dimeric coiled-coil: the Acanthamoeba myosin II rod. AB - Acanthamoeba myosin II rod is a long alpha-helical coiled-coil with a flexible hinge containing a helix-breaking proline. The thermal stability of the complete rod domain of myosin II (residues 849-1509), a mutant in which the hinge proline was replaced by alanine (P398A), and a mutant with the whole hinge region deleted (delta(384-408)) was studied in 0.6 and 2.2 M KCl, pH 7.5. In analytical ultracentrifugation studies, the purified myosin II rods sedimented as monodisperse dimers with sedimentation coefficients s(20,w) = 3.8 S (wild-type, Mr = 149,000) and 3.6 S (P398A and delta(384-408)). Circular dichroism (CD) and differential scanning calorimetry (DSC) showed that the thermal unfolding of the myosin II rod is reversible and highly cooperative. The unfolding of the rod is coupled to a dissociation of the chains, as shown by HPLC gel filtration at high temperatures and by the concentration dependence of the transition temperature. The CD and DSC data are consistent with a two-state mechanism (Tm approximately 40 degrees C, deltaH approximately 400 kcal/mol) in which the dimeric rod unfolds with concomitant formation of two unfolded monomers. We found no evidence for independent unfolding of the two rod domains that are separated by the hinge region. The only difference observed in the unfolding of the mutant rods from that of the wild type was a approximately 2 degrees C increase in the thermal stability of the hinge-deletion mutant. Thus, the mechanism of unfolding the Acanthamoeba myosin II rod is different from those of skeletal muscle myosin rod and tropomyosin, for which non-two-state thermal transitions have been observed. The cooperative unfolding of the entire coiled-coil rod of Acanthamoeba myosin II may underlie the previously reported regulatory coupling between its N-terminal head and C-terminal tail. PMID- 9201934 TI - Complex formation between the hepatitis C virus serine protease and a synthetic NS4A cofactor peptide. AB - The NS3 protein of the hepatitis C virus contains a serine protease that, upon binding to its cofactor, NS4A, is responsible for maturational cleavages that occur in the nonstructural region of the viral polyprotein. We have studied in vitro complex formation between the NS3 protease domain expressed in Escherichia coli and a synthetic peptide spanning the minimal domain of the NS4A cofactor. Complex dissociation constants in the low micromolar range were measured using different techniques such as activity titration, fluorescence titration, and pre equilibrium analysis of complex formation. Cofactor binding was strictly dependent on the glycerol content of buffer solutions and was not significantly influenced by substrate saturation of the enzyme. NS4A peptide binding to NS3 was accompanied by changes in the circular dichroism spectrum in the region between 270 and 290 nm, as well as by an enhancement of tryptophan fluorescence. Conversely, no changes in the far UV region of the circular dichroism spectrum were detectable. These data are indicative of induced tertiary structure changes and suggest that the secondary structure content of the uncomplexed enzyme does not differ significantly from that of the NS3-cofactor complex. Pre-equilibrium measurements of complex formation showed very low values for k(on), suggesting conformational transitions to be rate limiting for the association reaction. PMID- 9201933 TI - How enzymes control the reactivity of adenosylcobalamin: effect on coenzyme binding and catalysis of mutations in the conserved histidine-aspartate pair of glutamate mutase. AB - Glutamate mutase is one of a group of adenosylcobalamin-dependent enzymes that catalyze unusual isomerizations that proceed through the formation of radical intermediates. It shares a structurally similar cobalamin-binding domain with methylcobalamin-dependent methionine synthase. In particular, both proteins contain the "DXHXXG" cobalamin-binding motif, in which the histidine provides the axial ligand to cobalt. The effects of mutating the conserved histidine and aspartate residues in methionine synthase have recently been described [Jarrett, J. T., Amaratunga, M., Drennan, C. L., Scholten, J. D., Sands, R. H., Ludwig, M. L., & Matthews, R. G. (1996) Biochemistry 35, 2464-2475]. Here, we describe how similar mutations in the "DXHXXG" motif of glutamate mutase affect coenzyme binding and catalysis in an adenosylcobalamin-dependent reaction. The mutations made in the MutS subunit of glutamate mutase were His16Gly, His16Gln, Asp14Asn, Asp14Glu, and Asp14Ala. All the mutations affect, in varying degrees, the rate of catalysis, the affinity of the protein for the coenzyme, and the coordination of cobalt. Mutations of either Asp14 or His16 decrease k(cat) by 1000-fold, and whereas cob(II)alamin accumulates as an intermediate in the wild-type enzyme, it does not accumulate in the mutants, suggesting the rate-determining step is altered. The apparent Kd for adenosylcobalamin is raised by about 50-fold when His16 is mutated and by 5-10-fold when Asp16 is mutated. There are extensive differences between the UV-visible spectra of wild-type and mutant holoenzymes, indicating that the mutant enzymes coordinate cobalt less well. Overall, the properties of these mutants differ quite markedly from those observed when similar mutations were introduced into methionine synthase. PMID- 9201935 TI - Denaturation and self-association of apolipoprotein A-I investigated by electrophoretic techniques. AB - Purified human apolipoprotein A-I (apoA-I), when run across a transverse urea gradient at alkaline pH, gives a complex pattern characterized by a number of parallel sigmoidal curves, in which the transition between high- and low-mobility forms, i.e. from folded to unfolded structure, occurs between 1.1 and 3.2 M urea. Size differences appear to be the major cause of this isomerism. When migrated across a wide pH range in the presence of varying amounts of urea to display its titration curve, apoA-I is resolved into two pairs of bands, running parallel in the neutral to basic pH region while merging at acidic pH; such a finding does not correlate with a differential exposure of His residues, as shown by diethyl pyrocarbonate titration. Ferguson plot analysis, confirmed by cross-linking experiments, demonstrates a gradual shift from higher to lower mass aggregates as the urea concentration is raised; the monomeric form undergoes denaturation by swelling to an approximately 50% larger hydrodynamic volume than in its native state. At alkaline pH, where apoA-I exists as aggregated species, disaggregation and unfolding appear to happen at once, the larger aggregates being less stable than the smaller ones. At acidic pH, apoA-I does not form aggregates and has little secondary structure; unfolding is then a progressive rather than a cooperative process. PMID- 9201936 TI - Nonlamellar phases induced by the interaction of gramicidin S with lipid bilayers. A possible relationship to membrane-disrupting activity. AB - The interactions of the cyclic peptide gramicidin S (GS) with a variety of single component lipid bilayers, and with membrane polar lipid extracts of Acholeplasma laidlawii B and Escherichia coli, were examined by differential scanning calorimetry (DSC), 31P-nuclear magnetic resonance (NMR) spectroscopy, and X-ray diffraction. The DSC data indicate that the effects of GS on the thermotropic phase behavior of phosphatidylcholine and phosphatidylethanolamine dispersions are compatible with those expected of peptides interacting primarily with the polar headgroup and/or the polar/apolar interfaces of lipid bilayers. These DSC studies also suggest that GS exhibits stronger interactions with the more fluid bilayers. For mixtures of GS with lipids such as phosphatidylcholine, phosphatidylserine, cardiolipin, and sphingomyelin, axially symmetric 31P-NMR powder patterns are observed throughout the entire temperature range examined (0 90 degrees C), and there is little evidence for significant destabilization of the lipid bilayer with respect to nonlamellar phases. With mixtures of GS with either phosphatidylethanolamine, phosphatidylglycerol, or a nonlamellar phase forming phosphatidylcholine, axially symmetric 31P-NMR powder patterns are also observed at low temperatures. However, at high temperatures, an isotropic component is observed in their 31P-NMR spectra, and the relative intensity of this component increases significantly with temperature and with GS concentration. Once formed at high temperatures, this isotropic component exhibits a marked cooling hysteresis and in most cases disappears only when the sample is recooled to temperatures well below the lipid hydrocarbon chain-melting phase transition temperature. We also show that GS induces the formation of isotropic components in the 31P-NMR spectra of heterogeneous lipid mixtures such as occur in A. laidlawii B and E. coli membranes. These observations suggest that GS induces the formation of cubic or other three dimensionally ordered inverted nonlamellar phases when it interacts with some types of lipid bilayers, a suggestion strongly supported by our X-ray diffraction studies. Our results also suggest that the capacity of GS to induce the formation of such phases increases with the intrinsic nonlamellar phase-preferring tendencies of the lipids with which it interacts probably by producing localized increases in membrane monolayer curvature stress. The latter effect could be part of the mechanism through which this peptide exhibits its antimicrobial and hemolytic activities. PMID- 9201938 TI - Examination of the transition state of the low-molecular mass small tyrosine phosphatase 1. Comparisons with other protein phosphatases. AB - The reactions of p-nitrophenyl phosphate (pNPP) with the low-molecular mass tyrosine phosphatase Stp1 and with the mutants D128N, D128A, D128E, and S18A have been studied by measurement of heavy-atom isotope effects in the substrate. The isotope effects were measured at the nonbridging oxygen atoms [18(V/K)nonbridge], at the bridging oxygen atom (the site of bond cleavage) [18(V/K)bridge], and at the nitrogen atom in the nitrophenol leaving group [15(V/K)]. The results with native Stp1 were 1.0160 +/- 0.0005 for 18(V/K)bridge, 1.0007 +/- 0.0001 for 15(V/K), and 1.0018 +/- 0.0003 for 18(V/K)nonbridge. The values for 18(V/K)nonbridge and 15(V/K) differ from those previously measured with other protein-tyrosine phosphatases and from those of the aqueous hydrolysis reaction of pNPP. The values indicate that in the transition state of the native Stp1 reaction the leaving group bears a partial negative charge, and there is nucleophilic interaction between the Cys nucleophile, and the phosphoryl group, causing some decrease in the nonbridge P-O bond order. The transition state remains highly dissociative with respect to the degree of bond cleavage to the leaving group. Mutation of the general acid from aspartic acid to glutamic acid slows catalysis but causes no change in the isotope effects and thus does not alter the degree of proton transfer to the leaving group in the transition state. Mutations of this residue to asparagine or alanine give values for 18(V/K)bridge of about 1.029, for 15(V/K) of about 1.003, and for 18(V/K)nonbridge of 1.0010 (D128A) to 1.0024 (D128N). These data indicate a dissociative transition state with the leaving group departing as the nitrophenolate anion and indicate more nucleophilic participation than in the aqueous hydrolysis of the pNPP dianion, just as in the native enzyme. The isotope effects with the S18A mutant, in which a hydrogen bonding stabilization of the anionic Cys nucleophile has been removed, were within experimental error of those with the native enzyme, indicating that this alteration has no effect on the transition state for phosphoryl transfer from pNPP. PMID- 9201937 TI - Rapid and parallel formation of Fe3+ multimers, including a trimer, during H-type subunit ferritin mineralization. AB - Conversion of Fe ions in solution to the solid phase in ferritin concentrates iron required for cell function. The rate of the Fe phase transition in ferritin is tissue specific and reflects the differential expression of two classes of ferritin subunits (H and L). Early stages of mineralization were probed by rapid freeze-quench Mossbauer, at strong fields (up to 8 T), and EPR spectroscopy in an H-type subunit, recombinant frog ferritin; small numbers of Fe (36 moles/mol of protein) were used to increase Fe3+ in mineral precursor forms. At 25 ms, four Fe3+-oxy species (three Fe dimers and one Fe trimer) were identified. These Fe3+ oxy species were found to form at similar rates and decay subsequently to a distinctive superparamagentic species designated the "young core." The rate of oxidation of Fe2+ (1026 s(-1)) corresponded well to the formation constant for the Fe3+-tyrosinate complex (920 s(-1)) observed previously [Waldo, G. S., & Theil, E. C. (1993) Biochemistry 32, 13261] and, coupled with EPR data, indicates that several or possibly all of the Fe3+-oxy species involve tyrosine. The results, combined with previous Mossbauer studies of Y30F human H-type ferritin which showed decreases in several Fe3+ intermediates and stabilization of Fe2+ [Bauminger, E. R., et al. (1993) Biochem. J. 296, 709], emphasize the involvement of tyrosyl residues in the mineralization of H-type ferritins. The subsequent decay of these multiple Fe3+-oxy species to the superparamagnetic mineral suggests that Fe3+ species in different environments may be translocated as intact units from the protein shell into the ferritin cavity where the conversion to a solid mineral occurs. PMID- 9201939 TI - Does poly(ADP-ribosyl)ation regulate the DNA methylation pattern? AB - The existence of a possible correlation between poly(ADP-ribosyl)ation and DNA methylation processes was investigated. In vivo and in vitro experiments were carried out on L929 mouse fibroblasts preincubated for 24 h with or without 3 aminobenzamide, a well-known inhibitor of poly(ADP-ribose) polymerase. Both experimental approaches evidenced a close relationship between these two important nuclear enzymatic mechanisms, suggesting that the poly(ADP-ribosyl)ated isoform of H1 histone and/or long and branched protein-free ADP-ribose polymers could act as protecting agents against full methylation of the CpG dinucleotides in genomic DNA. PMID- 9201941 TI - Cleavage of 16S rRNA within the ribosome by mRNA modified in the A-site codon with phenanthroline-Cu(II). AB - Cleavage of 16S rRNA was obtained through mRNA modified at position +5 with the chemical cleavage agent 1,10-o-phenanthroline. In the presence of Cu2+, and after addition of reducing agent to the modified mRNA-70S complex, cleavage of proximal nucleotides within the 16S rRNA occurred. Primer extension analysis of 16S rRNA fragments revealed that nucleotides 528-532, 1196, and 1396-1397 were cleaved. Nucleotides 1053-1055 were also cleaved but did not show the same level of specificity as the former. These results provide evidence that at some point in the translation process these regions are all within 15 A of position +5, the A site codon, on the mRNA. PMID- 9201940 TI - Mutational analysis of 26 residues of vaccinia DNA topoisomerase identifies Ser 204 as important for DNA binding and cleavage. AB - Vaccinia DNA topoisomerase, a 314 amino acid type I enzyme, catalyzes the cleavage and rejoining of DNA strands through a DNA-(3'-phosphotyrosyl)-enzyme intermediate formed at a specific target sequence, 5'-(C/T)CCTT downward arrow. To identify amino acids that participate in the DNA binding and transesterification steps, we introduced alanine substitutions at 18 positions within a centrally located 27 amino acid segment (181-RLYKPLLKLTDDSSPEEFLFNKLSERK 207) and at 8 positions near the N-terminus (1-MRALFYKDGK-10). All mutant proteins except two displayed wild-type activity in relaxing supercoiled DNA. F200A and S204A exhibited reduced rates of relaxation and were subjected to a kinetic analysis of the strand cleavage reaction under single-turnover and equilibrium conditions. The F200A and S204A mutations reduced the rate of single turnover DNA cleavage by factors of 5 and 70, respectively. Both mutations shifted the cleavage-religation equilibrium in favor of the noncovalently bound state. The S204A mutation reduced the affinity of topoisomerase for CCCTT containing DNA, but did not alter the site-specificity of DNA cleavage. Vaccinia residue Ser-204, which is conserved in all poxvirus topoisomerases, but not in the cellular homologues, may contribute to the unique cleavage site specificity of the poxvirus enzymes. Phe-200 is conserved in all members of the type IB topoisomerase family. PMID- 9201942 TI - Structural characterization of Paracoccus denitrificans cytochrome c peroxidase and assignment of the low and high potential heme sites. AB - The amino acid sequence of the diheme cytochrome c peroxidase from Paracoccus denitrificans has been determined as the result of sequence analysis of peptides generated by chemical and enzymatic cleavages of the apoprotein. The sequence shows 60% similarity to the cytochrome c peroxidase from Pseudomonas aeruginosa, 39% similarity to an open reading frame encoding a putative triheme c-type cytochrome in Escherichia coli, and remote similarity to the MauG proteins from two methylotrophic bacteria. It is proposed, on the basis of the pattern of conserved residues in the sequences, that a change in iron coordination in the N terminal heme domain may accompany reduction to the active mixed valence state, a change which may be accompanied by conformational adjustments in the highly conserved interface between the N- and C-terminal domains. These conformational adjustments may also lead to the appearance of a second Ca2+ binding site in the mixed valence enzyme. The exposed edge of the heme in the C-terminal domain is surrounded by several different patterns of charged residues in the Paracoccus and Pseudomonas enzymes, and this is consistent with the interaction of the former with the highly positively charged front face of the donor cytochrome c 550. PMID- 9201943 TI - An RNA structural determinant for tRNA recognition. AB - Escherichia coli tRNACys contains an unusual G15.G48 tertiary base pair that is important for recognition and aminoacylation by cysteine tRNA synthetase. This G15.G48 tertiary base pair has a distinctive chemical modification signature that suggests an N2.N3 base pairing. The N2.N3 pairing of a G.G base pair has not been described in any existing RNA structures. Identification of the structural determinant of G15.G48 is of fundamental importance for understanding the formation of an RNA tertiary base pair, as well as the role of RNA tertiary structure in tRNA recognition. We show here that the structural determinant for G15.G48 is an A13.A22 mismatch in the dihydrouridine stem. Introduction of A13.A22 to an unrelated tRNA confers the distinctive chemical modification signature of G15. G48 while substitution of A13.A22 eliminates this signature. The relationship between G15.G48 and A13.A22 enables the unrelated tRNA to be efficiently recognized by cysteine tRNA synthetase. Modeling studies show that A13.A22 has the potential to form a base triple with A46, which is directly connected to G48 in the G15.G48 base pair. The proposed A13.A22.A46 base triple provides a framework for understanding how two RNA structural elements may be related to each other in playing an important role in tRNA aminoacylation. PMID- 9201944 TI - Extended DNA-recognition repertoire of peptide nucleic acid (PNA): PNA-dsDNA triplex formed with cytosine-rich homopyrimidine PNA. AB - Peptide nucleic acid (PNA) is an oligonucleotide mimic in which the backbone of DNA has been replaced by a pseudopeptide. Thymine-rich homopyrimidine PNA oligomers have been found to recognize double-stranded DNA targets by displacement of the pyrimidine DNA strand and forming an internal Watson-Crick Hoogsteen base-paired PNA(pyr)-DNA(pu)-PNA(pyr) triplex. We here show that cytosine-rich homopyrimidine PNA sequences instead add to double-stranded polynucleotide targets as Hoogsteen strands forming PNA(pyr)-DNA(pu)-DNA(pyr) triplexes. Furthermore, PNA strands with homopurine or alternating thymine guanine sequences are shown to invade their respective DNA targets by displacing the identical DNA strands of the polynucleotides and forming new PNA-DNA duplexes. These results indicate distinct mechanistic variations as to how PNA interacts with a DNA target depending on choice of nucleobases, which could be of importance for future design of gene-specific diagnostic or therapeutic agents. PMID- 9201946 TI - Kinetics of the RNA-DNA helicase activity of Escherichia coli transcription termination factor rho. 2. Processivity, ATP consumption, and RNA binding. AB - The RNA-binding and RNA-DNA helicase activities of the Escherichia coli transcription termination factor rho have been investigated using natural RNA molecules that are 255 and 391 nucleotide residues in length and that contain the trp t' rho-dependent termination sequence of E. coli. Helicase substrates were prepared from these RNA molecules by annealing one or more DNA oligomers to complementary sequences located at or near the 3'-ends of the RNA molecules to form defined RNA-DNA hybrid sequences ranging in length from 20 to 100 bp. By comparing the fraction of the RNA molecules bound to rho with the fraction of bound DNA oligomers removed from the RNA during one round of the helicase reaction, we have shown that rho translocates processively at 37 degrees C in buffer containing 50 mM KCl. Helicase reactions and ATPase measurements were performed in parallel in the presence of RNA molecules containing RNA-DNA hybrids of various lengths, and we show that both the rate of translocation of the rho hexamer along the RNA chain and the rate of ATP consumption are similar, whether or not DNA is hybridized to the RNA transcript. By combining measurements of translocation and ATPase rates, we estimate that rho consumes approximately 1-2 ATP molecules in translocating over 1 nucleotide residue of the RNA chain at 37 degrees C in 50 mM KCl. The ATPase activity of rho remains the same after one round of the helicase reaction, indicating that rho appears to hydrolyze ATP at the same rate, whether it is translocating along the RNA, separating RNA-DNA hybrids, or bound at the 3'-end of the RNA substrate. We also show that rho binds cooperatively ( approximately 2-4 rho hexamers per RNA chain) to the RNA substrates under our standard helicase reaction conditions. However, cooperative binding is not essential for helicase activity, since this binding stoichiometry can be reduced to approximately 1.5 rho hexamers per 255-nucleotide residue RNA chain by blocking approximately 100 nt of either end of the rho binding site of the helicase substrate with complementary DNA oligonucleotides, with no change in helicase properties. The implications of these results for models of rho helicase function and for the role of rho in termination are discussed. PMID- 9201945 TI - Kinetics of the RNA-DNA helicase activity of Escherichia coli transcription termination factor rho. 1. Characterization and analysis of the reaction. AB - The kinetics of the ATP-dependent RNA-DNA helicase activity of Escherichia colitranscription termination factor rho have been analyzed. Helicase substrates were assembled using 255 nt and 391 nt RNA sequences from the trp t' RNA transcript of E. coli. These RNA sequences each carry a rho "loading site" at a position near the 5'-end, and a rho-dependent terminator sequence at the 3'-end to which complementary approximately 20 nt DNA oligonucleotides have been annealed. A rapid ( approximately 30 s) pre-steady-state burst of helicase activity (DNA oligomer release), followed by a slow linear phase, is observed in reactions carried out at low salt concentrations (50 mM KCl). Using poly(rC) or poly(dC) as traps for the rho that is released after one round of activity, we have shown that the first (burst) phase of the reaction represents the processive translocation of prebound rho hexamers from the rho loading site to the 3'-end of the RNA molecule. The slow phase of the reaction is complex and represents a combination of many different processes, including the slow release of RNA from rho, the reannealing of complementary DNA oligonucleotides to the RNA substrate, and the recycling of rho hexamers onto additional RNA molecules. Reactions carried out at higher salt concentrations (150 mM KCl) consist of only one phase, since under these conditions rho dissociates more rapidly from the RNA, with an amplitude corresponding to several DNA oligomers removed per rho hexamer. Thus, rho can recycle and function as a catalytic helicase under reaction conditions resembling those found in the cell. PMID- 9201947 TI - Spectroscopic determination of open complex formation at promoters for Escherichia coli RNA polymerase. AB - A considerable amount of effort has been expended studying the kinetics of association of Escherichia coli RNA polymerase with promoter DNA in forming the open complex. Strand separation occurs over about 12 base pairs and includes the transcription start site. However, these efforts have been significantly hampered by the lack of a sensitive, real time method by which formation of an open complex could be assayed. Here, we employ short (86 bp) synthetic promoters with 2-aminopurine (2-AP) substitutions in the region that becomes single-stranded to spectroscopically monitor open complex formation. We demonstrate that promoters bearing the substitutions behave in a manner similar to that of those containing only the four common bases with respect to both the region of strand separation and start site selection. Open complex formation was found to yield an increased fluorescence signal with an emission maximum characteristic of 2-aminopurine. This spectroscopic assay for open complex formation was found to be well-suited to the investigation of a strong promoter, allowing open complex formation to be followed over a time scale of seconds with a stopped flow apparatus. The introduction of two additional nonconsensus base pairs in the -35 region resulted in a promoter for which open complex formation was 100-fold slower. The same substrates were also used to monitor the promoter re-annealing that ensues upon initiation of RNA synthesis. Similar rates for this process were observed for the two promoter variants employed in this study. PMID- 9201948 TI - Deglycosylation susceptibility and base-pairing stability of 2'-deoxyoxanosine in oligodeoxynucleotide. AB - We have demonstrated recently that nitrous acid or nitric oxide converts 2' deoxyguanosine (dGuo) into 2'-deoxyoxanosine (dOxo) [Suzuki, T., Yamaoka, R., Nishi, M., Ide, H., & Makino, K. (1996) J. Am. Chem. Soc. 118, 2515-2516]. In the present study, we have measured susceptibility of the N-glycosidic bond of dOxo to spontaneous hydrolysis and its base-pairing stability to evaluate the biological significance of dOxo as a new lesion in DNA. When oligodeoxynucleotide d(T5OT6) (O = dOxo), isolated from nitrous acid-treated d(T5GT6), was incubated at pH 4.0 and 70 degrees C, hydrolysis of the N-glycosidic bond of dOxo occurred with a first-order rate constant. Comparison of the rate constants with those of dGuo and dXao indicates that the N-glycosidic bond of dOxo was as stable as that of dGuo in d(T5GT6) and hydrolyzed 44-fold more slowly than that of 2' deoxyxanthosine (dXao), a simultaneously generated damage by nitrous acid and nitric oxide. For the estimation of the base-pairing stability, UV melting curves were measured for the duplexes of d(T5OT6).d(A6NA5) (N = A, G, C, and T) at neutral pH. The Tm values obtained were 15.3, 14.1, 19.3, and 16. 3 degrees C for N = A, G, C, and T, respectively, which are much lower than that of the intact duplex containing a G.C pair at the same position [d(T5GT6).d(A6CA5), Tm = 32.8 degrees C] but comparable with those of d(T5XT6).d(A6NA5) (X = dXao, Tm = 14.8 22.3 degrees C). CD spectra of the four duplexes containing dOxo showed preservation of the structure of the intact duplex at low temperature. UV and NMR pH-titration studies indicated the pKa for the ring-opening and -closing equilibrium to be 9.4, implying that dOxo is in the ring-closed form at physiological pH. This structure appears to be not suitable geometrically for the hydrogen bond formation with a specific counter base, thus causing equally low Tm values for all the counter bases. Consequently, these results imply that dOxo, a novel DNA lesion, may have an important and unique role in mutagenic events in cells. PMID- 9201949 TI - Oligomerization of the amide sensor protein AmiC by x-ray and neutron scattering and molecular modeling. AB - AmiC is the negative regulator of the amidase operon which is involved in amide metabolism in the cytosol of Pseudomonas aeruginosa. Crystal structures show that AmiC contains two large domains that are very similar to the periplasmic leucine isoleucine-valine binding protein (LivJ) of Escherichia coli. Synchrotron X-ray and neutron (in 100% 2H2O buffer) scattering data were obtained for AmiC in the presence of its substrate acetamide and its anti-inducer butyramide which binds more weakly to AmiC than acetamide. Guinier analyses to obtain radius of gyration RG and molecular weight Mr values showed that AmiC formed trimers whose formation was favored in the presence of acetamide and which exhibited concentration dependent properties at concentrations between 0.4 and 2 mg/mL. Above 2 mg/mL, where trimers predominated, the RG data were identical within 0.05 nm for AmiC acetamide and AmiC-butyramide with mean X-ray and neutron RG values of 3.35 and 3. 28 nm, respectively. Scattering curve fits constrained by the crystal structure of AmiC-acetamide were evaluated in order to describe a model for trimeric AmiC. A translational search of parallel alignments of three monomers to form a symmetric AmiC homotrimer gave a good X-ray curve fit. Combinations of calculated curves for monomeric, dimeric, trimeric, and tetrameric AmiC as seen in the crystal structure of AmiC gave reasonable but weaker X-ray curve fits which did not favor the existence of tetrameric AmiC. It is concluded that AmiC exhibits novel ligand-dependent oligomerization properties in solution when these are compared to other members of the periplasmic binding protein superfamily, where AmiC exists in monomeric and trimeric forms, the proportions of which depend on the presence of acetamide or butyramide. PMID- 9201950 TI - Investigation of the structural basis for thermodynamic stabilities of tandem GU wobble pairs: NMR structures of (rGGAGUUCC)2 and (rGGAUGUCC)2. AB - The symmetric, tandem GU mismatch motifs, and , which only differ in the mismatch order, have an average difference in thermodynamic stability of 2 kcal/mol at 37 degrees C. Thermodynamic studies of duplexes containing these motifs indicate the effect is largely localized to the mismatches and adjacent base pairs. The three dimensional structures of two representative duplexes, (rGGAGUUCC)2 and (rGGAUGUCC)2, were determined by two-dimensional NMR and a simulated annealing protocol. Local deviations are similar to other intrahelical GU mismatches with little effect on backbone torsion angles and a slight overtwisting between the base pair 5' of the G of the mismatch and the mismatch itself. Comparisons of the resulting stacking patterns along with electrostatic potential maps suggest that interactions between highly negative electrostatic regions between base pairs may play a role in the observed thermodynamic differences. PMID- 9201951 TI - Partial activation of muscle phosphorylase by replacement of serine 14 with acidic residues at the site of regulatory phosphorylation. AB - Phosphorylation of glycogen phosphorylase at residue Ser14 triggers a conformational transition that activates the enzyme. The N-terminus of the protein, in response to phosphorylation, folds into a 310 helix and moves from its location near a cluster of acidic residues on the protein surface to a site at the dimer interface where a pair of arginine residues form charged hydrogen bonds with the phosphoserine. Site-directed mutagenesis was used to replace Ser14 with Asp and Glu residues, analogs of the phosphoserine, that might be expected to participate in ionic interactions with the arginine side chains at the dimer interface. Kinetic analysis of the mutants indicates that substitution of an acidic residue in place of Ser14 at the site of regulatory phosphorylation partially activates the enzyme. The S14D mutant shows a 1.6-fold increase in Vmax, a 10-fold decrease in the apparent dissociation constant for AMP, and a 3 fold decrease in the S0.5 for glucose 1-phosphate. The S14E mutant behaves similarly, showing a 2.2-fold increase in Vmax, a 6-fold decrease in the apparent dissociation constant for AMP, and a 2-fold decrease in the S0.5 for glucose 1 phosphate. The ability of the mutations to enhance binding of AMP and glucose 1 phosphate and to raise catalytic activity suggests that the introduction of a carboxylate side chain at position 14 promotes docking of the N-terminus at the subunit interface and concomitant stabilization of the activated conformation of the enzyme. Like the native enzyme, both mutants show significant activity only in the presence of the activator, AMP. Full activation, analogous to that provided by covalent phosphorylation of the enzyme, likely is not achieved because of differences in the charge and the geometry of ionic interactions at the phosphorylation site. PMID- 9201953 TI - Solution conformations and interactions of alpha and beta subunits of the Oxytricha nova telomere binding protein: investigation by Raman spectroscopy. AB - Solution conformations of the alpha and beta subunits of the Oxytricha nova telomere binding protein have been investigated by Raman spectroscopy. Raman spectra have also been obtained for a deletion mutant of the beta subunit, betaC232, which retains the N-terminal domain that is active in ternary complex (alpha:beta:DNA) formation but lacks the C-terminal domain that is active in catalyzing guanine quadruplex formation. The Raman spectra show that alpha, beta, and betaC232 are rich in beta-strand secondary structure ( approximately 40-50%) and turns. The Raman signature of the C-terminal 153 amino acids of beta, generated by subtracting the spectrum of betaC232 (residues 1-232) from that of the full subunit, indicates that the domain active in guanine quadruplex formation contains less beta-strand secondary structure and more irregular structure than the domain active in alpha:beta:DNA formation. Raman markers also provide information about the environments and orientations of several key side chains, including tryptophan residues in N- and C-terminal domains of the beta subunit. Both alpha and beta denature between 30 and 40 degrees C, as evidenced by large changes in Raman bands diagnostic of main chain conformation and side chain environments. The Raman spectrum of an equimolar alpha/beta mixture exhibits no evidence of specific interaction between the subunits; further, the denaturation profile of this mixture is indistinguishable from the sum of denaturation profiles of the constituent subunits, consistent with the absence of appreciable interaction between alpha and beta throughout the range 0-50 degrees C. The present results provide insights into the solution conformations of the Oxytricha telomere binding protein subunits and serve as the basis for future study of subunit interactions with telomeric DNA. PMID- 9201952 TI - A proposal for the Mg2+ binding site of P-type ion motive ATPases and the mechanism of phosphoryl group transfer. AB - Mutations of D586 in the DPPR sequence of sodium pump decrease the enzyme's affinity for inorganic phosphate [Farley R. A., Heart, E., Kabalin, M., Putnam, D., Wang, K., Kasho, V. N., and Faller, L. D. (1997) Biochemistry 36, 941-951]. Therefore, it was proposed that D586 coordinates the Mg2+ required for catalytic activity. This hypothesis is tested (1) by determining the substrate for catalysis of 18O exchange between inorganic phosphate and water and (2) by comparing conserved amino acid sequences in P-type pumps with the primary structures of enzymes of known tertiary structure that catalyze phosphoryl group transfer. From the isotope exchange data, it is concluded that the Mg2+-dependent and Na+- and K+-stimulated ATPase binds Mg2+ before inorganic phosphate. Sequence homology is demonstrated between the conserved DPPR and MV(I,L)TGD sequences of P type pumps and two conserved adenylate kinase sequences that coordinate Mg2+ and/or bind nucleotide in the crystal structure of the yeast enzyme. A model for the Mg2+ site of P-type pumps and the mechanism of phosphoryl group transfer is proposed and tested by demonstrating that the conserved sequences are also structurally homologous. PMID- 9201954 TI - Unusual mechanism of oxygen atom transfer and product rearrangement in the catalytic reaction of 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase. AB - The oxygenation reaction of 2-methyl-3-hydroxypyridine-5-carboxylic acid (MHPC) oxygenase with the substrate, MHPC, was investigated. Two oxygenated flavin intermediates C(4a)-hydroperoxy flavin and C(4a)-hydroxy flavin were found, implying that the enzyme functions similarly to flavoprotein hydroxylases. This finding is supported by the results of independent oxygen-18 tracer experiments, which showed that one atom of oxygen from 18O2 and one atom of oxygen from H218O are incorporated in the product. MHPC oxygenase normally catalyzes both the oxygenation and the hydrolytic ring opening of the pyridine ring of MHPC to yield the acyclic compound, alpha-(N-acetylaminomethylene)succinic acid. Using 5 hydroxynicotinic acid (5HN), which has no 2-methyl group, we tested whether the hydrolytic reaction was due to the presence of the 2-methyl group on MHPC (that prevented rearomatization of the initial product) or to the specific properties of MHPC oxygenase. Product analysis of the enzymatic reaction of 5HN and MHPC oxygenase shows that the enzyme catalyzes the hydroxylation and subsequent hydrolysis of the hydroxylated substrate to yield an acyclic product. The investigation of the oxygenation reaction demonstrates that the enzyme uses the same mechanism to catalyze the 5HN reaction as it does in the MHPC reaction. PMID- 9201955 TI - 2-Oxo-3-alkynoic acids, universal mechanism-based inactivators of thiamin diphosphate-dependent decarboxylases: synthesis and evidence for potent inactivation of the pyruvate dehydrogenase multienzyme complex. AB - A new class of compounds, the 2-oxo-3-alkynoic acids with a phenyl substituent at carbon 4 was reported by the authors as potent irreversible and mechanism-based inhibitors of the thiamin diphosphate- (ThDP-) dependent enzyme pyruvate decarboxylase [Chiu, C.-F., & Jordan, F. (1994) J. Org. Chem. 59, 5763-5766]. The method has been successfully extended to the synthesis of the 4-, 5-, and 7 carbon aliphatic members of this family of compounds. These three compounds were then tested on three ThDP-dependent pyruvate decarboxylases: the Escherichia coli pyruvate dehydrogenase multienzyme complex (PDHc) and its E1 (ThDP-dependent) component, pyruvate oxidase (POX, phosphorylating; from Lactobacillus plantarum),and pyruvate decarboxylase (PDC) from Saccharomycescerevisiae. All three enzymes were irreversibly inhibited by the new compounds. The 4-carbon acid is the best substrate-analog inactivator known to date for PDHc, more potent than either fluoropyruvate or bromopyruvate. The following conclusions were drawn from extensive studies with PDHc: (a) The kinetics of inactivation of PDH complexes and of resolved E1 by 2-oxo-3-alkynoic acids is time- and concentration dependent. (b) The 4-carbon acid has a Ki 2 orders of magnitude stronger than the 5-carbon acid, clearly demonstrating the substrate specificity of PDHc. (c) The rate of inactivation of PDH complexes and of resolved E1 by 2-oxo-3-alkynoic acids is enhanced by the addition of ThDP and MgCl2. (d) Pyruvate completely protects E1 and partially protects PDHc from inactivation by 2-oxo-3-butynoic acid. (e) E1 but not E2-E3 is the target of inactivation by 2-oxo-3-butynoic acid. (f) Inactivation of E1 by 2-oxo-3-butynoic acid is accompanied by modification of 1.3 cysteines/E1 monomer. The order of reactivity with the 4 carbon acid was PDHc > POX > PDC. While the order of reactivity with PDHc and POX was 2-oxo-3-butynoic acid > 2-oxo-3-pentynoic acid > 2-oxo-3-heptynoic acid, the order of reactivity was reversed with PDC. PMID- 9201956 TI - Cobalamin-dependent methionine synthase is a modular protein with distinct regions for binding homocysteine, methyltetrahydrofolate, cobalamin, and adenosylmethionine. AB - Methionine synthase (MetH) catalyzes the transfer of a methyl group from bound methylcobalamin to homocysteine, yielding enzyme-bound cob(I)alamin and methionine. The cofactor is then remethylated by methyltetrahydrofolate. We now demonstrate that MetH is able to catalyze methylation of free cob(I)alamin with methyltetrahydrofolate. MetH had previously been shown to catalyze methylation of homocysteine with free methylcobalamin as the methyl donor, in a reaction that is first-order in added methylcobalamin, and we have confirmed this observation using homogenous enzyme. A truncated polypeptide lacking the cobalamin-binding region of the holoenzyme, MetH(2-649), was overexpressed and purified to homogeneity. MetH(2-649) catalyzes the methylation of free cob(I)alamin by methyltetrahydrofolate and the methylation of homocysteine by free methylcobalamin. Furthermore, a protein comprising residues 2-353 of the holoenzyme has now been overexpressed and purified to homogeneity, and this protein catalyzes methyl transfer from free methylcobalamin to homocysteine but not from methyltetrahydrofolate to free cob(I)alamin. The mutations Cys310Ala and Cys311Ala in MetH(2-649) completely abolish methyl transfer from exogenous methylcobalamin to homocysteine but do not affect methyl transfer from methyltetrahydrofolate to exogenous cob(I)alamin, consistent with a modular construction for MetH. We infer that MetH is a modular protein comprising four separate regions: a homocysteine binding region (residues 2-353), a methyltetrahydrofolate binding region (residues 354-649), a region responsible for binding the cobalamin prosthetic group (residues 650-896), and an AdoMet binding domain (residues 897-1227). PMID- 9201957 TI - The carbohydrate recognition domain of surfactant protein A mediates binding to the major surface glycoprotein of Pneumocystis carinii. AB - Pneumocystis carinii is a common cause of life-threatening pneumonia in immunodeficient patients. Pulmonary surfactant protein A (SP-A), an alveolar glycoprotein containing collagen-like and carbohydrate recognition domains (CRD), binds P. carinii and enhances adherence to alveolar macrophages. In this study, we examined the structural basis of the interaction between SP-A and the major surface glycoprotein of P. carinii (MSG). Rat SP-A bound to purified rat P. carinii-derived MSG in a saturable and calcium-dependent manner, which was partially reversible by coincubation with excess monosaccharides, or pretreatment of MSG with N-glycanase. Mutant recombinant SP-As with neutral amino acid substitutions for the predicted calcium- and carbohydrate-coordinating residues of the CRD were synthesized in insect cells using baculovirus vectors and tested for binding to MSG. Substitutions of negatively charged (Glu195, Glu202, and Asp215) and polar residues (Asn214) of the CRD with alanine but not substitution of the Arg197 with glycine reduced the binding of SP-A to mannose-Sepharose beads and to MSG. Deletion of the N-linked oligosaccharides from SP-A by mutagenesis of the consensus sequences for glycosylation had no effect on binding. We conclude that the CRD mediates the binding of SP-A to oligosaccharides attached to MSG. PMID- 9201958 TI - Serine-578 is a major phosphorylation locus in human plasma plasminogen. AB - It has been reported that human plasminogen (HPg) exists in plasma in a phosphorylated form. We now document that both major glycoforms of plasma HPg contain a phosphoserine residue in their latent protease chains, as revealed by quantitative protein phosphate determinations and 31P-NMR analysis. The sequence location of the phosphoserine residue was established by time-of-flight matrix assisted laser desorption ionization with delayed extraction mass spectrometric analysis of peptides resulting from complete tryptic and cyanogen bromide digests of the latent protease chain of HPg. Confirmation of the presence of organic phosphate in the identified peptide was obtained by determination of the resulting mass shift after treatment of the peptide with alkaline phosphatase. The data show that Ser578 is a major phosphorylation site in HPg. PMID- 9201959 TI - C repeats of the streptococcal M1 protein achieve the human serum albumin binding ability by flanking regions which stabilize the coiled-coil conformation. AB - The M and M-like proteins of Streptococcus pyogenes are fibrous cell surface proteins. They have multiple binding sites for several human proteins and are composed of the C-terminal anchor domain, the alpha-helical coiled-coil domain, and the N-terminal non-coiled-coil domain. The coiled-coil domain of the M1 protein consists of repeat units called B, C, and D and a spacer unit S between B and C. Recombinant fragments A-B-S-C-D, A-B-S, B-S-C, S-C, S-C-D, C-D, and C of the coiled-coil domain were studied by analyzing their secondary structures and binding affinities to human serum albumin (HSA). As shown by circular dichroism, all fragments are in an alpha-helical conformation. C-D and S-C-D form coiled coils at room temperature and bind below 37 degrees C with high affinity to HSA. C-D and S-C-D unfold in two steps with Tm values of approximately 31 and approximately 65 degrees C; complex formation with HSA increases the unfolding temperatures. B-S-C has a lower alpha-helical content, a less pronounced coiled coil conformation, and a reduced thermal stability, binds HSA weaker, and is only slightly stabilized by HSA binding in comparison to C-D and S-C-D. C and S-C are less stable than the other fragments and are not organized as coiled coils showing some features of alpha-helical single strands only below 20 degrees C, and binding of HSA was not observed. The results indicate that the formation of coiled-coil structures, supported by flanking D regions and, to a lesser extent also B regions, is essential for the binding of C repeat units to HSA. PMID- 9201961 TI - Comparative Fourier transform infrared studies of the secondary structure and the CO heme ligand environment in cytochrome P-450cam and cytochrome P-420cam. AB - For the first time, Fourier transform infrared spectroscopy has been applied to cytochrome P-450 to analyze the protein secondary structure. From Fourier self deconvolution and fitting the infrared spectra in the amide I' region (1600-1700 cm-1), we estimate 44% alpha-helix, 31% beta-sheet, and 18% turns for substrate free cytochrome P-450cam. In the presence of camphor, 54% alpha-helix and 310 helix, 21% beta-sheet, and 21% turns are obtained which agree with the crystallographic data of 53% alpha-helix, 19% beta-sheet, and 16% turns [Poulos, T. L., Finzel, B. C., & Howard, A. J. (1987) J. Mol. Biol. 195, 687-700]. Cytochrome P-420cam is produced from substrate-free cytochrome P-450cam in two ways: (i) by temperature elevation up to 60 degrees C and (ii) by exposure to KSCN up to 1.5 M. The secondary structure composition is determined for each temperature and KSCN concentration and compared with the changes observed in the iron ligand CO stretch vibration bands appearing between 1900 and 2000 cm-1. Thermally induced cytochrome P-420 has an alpha-helix content of 19%, a beta sheet content of 53%, 14% turns, and 5% antiparallel beta-sheets from intermolecular hydrogen bonds within protein aggregates. The formation of cytochrome P-420 as a function of the KSCN concentration indicates two types of cytochrome P-420. Up to 1 M KSCN, the induced cytochrome P-420 displays only little modification of the secondary structure, whereas at 1.5 M KSCN, larger changes are observed, resulting in 85% cytochrome P-420 without protein precipitation and containing 30% alpha-helix, 48% beta-sheet, and 17% turns. Infrared spectra in the iron ligand CO stretch region show several subconformers for cytochrome P-420. During the cytochrome P-420 formation, the CO stretch modes are shifted to higher frequencies by 3-11 cm-1, with a main feature at about 1964 cm-1, compared to those of substrate-free cytochrome P-450cam-CO. PMID- 9201960 TI - Unique Alzheimer's disease paired helical filament specific epitopes involve double phosphorylation at specific sites. AB - Alzheimer's disease (AD) paired helical filaments (PHFs), building blocks of neurofibrillary tangles (NFTs) are composed of hyperphosphorylated forms of the microtubule-associated protein tau (i.e., PHF-tau). Currently, much effort is devoted to the development of diagnostic antibodies specific for PHF-tau since elevated tau levels are found in the cerebral spinal fluid of AD patients. To this end, we have mapped the epitopes of a large panel of monoclonal antibodies (mAbs) that recognized only phosphorylation dependent epitopes on PHF-tau. These mAbs include the PHF-tau specific mAb AT10 and 12 newly developed anti-PHF mAbs that recognize PHF-tau but not autopsy-derived normal adult tau on Western-blot and enzyme-linked immunosorbent assay (ELISA). Epitope analysis, together with data on known binding sites of previously published mAbs, revealed that Ser214, Thr231, and Ser396 are immunodominant phosphorylated amino acids in PHF-tau. Six of the 12 new mAbs recognized one of these three phosphorylated sites. With the exception of AT10 and PHF-27, all the mAbs also labeled fetal tau and biopsy derived tau. Since mAbs AT10 and PHF-27 had little or no affinity for fetal tau and biopsy tau, they can be considered as the first "true" PHF-specific antibodies capable of distinguishing tau isoforms from normal versus AD subjects, suggesting a possible utility of these mAbs as diagnostic markers. Remarkably, the true PHF-specific antibodies recognized peptide sequences phosphorylated on more than one amino acid residue. The peptide recognition of mAb AT10 required the simultaneous phosphorylation of Thr212 and Ser214, and the peptide recognition of mAb PHF-27 was markedly increased when both the primary site Thr231 and the subsite Ser235 were phosphorylated. Since AT10 and PHF-27 are the only mAbs currently available that bind specifically to PHF-tau, these data suggest that double phosphorylation at Thr212/Ser214 and Thr231/Ser235 may be unique to PHF-tau. These data may facilitate the development of mAbs that can be used as specific diagnostic reagents for the detection of altered tau in cerebrospinal fluid of AD patients. PMID- 9201962 TI - Effects of viscosity and temperature on the kinetics of the electron-transfer reaction between the triplet state of zinc cytochrome c and cupriplastocyanin. AB - This is a study of the effects of viscosity (in the range of 0.8-790 cP), of temperature (in the range of 260.7-307.7 K), and of ionic strength (in the range of 2.5-20.0 mM) on the kinetics of photoinduced electron-transfer reaction 3Zncyt/pc(II) --> Zncyt+/pc(I) within the electrostatic complex of zinc cytochrome c and cupriplastocyanin at pH 7.0. The unimolecular rate constant is kF. The apparent activation parameters DeltaH*, DeltaS*, and DeltaG* for this reaction were obtained in experiments with aqueous glycerol solutions having a constant composition. The interpolation of kF values obtained at the constant composition into the dependence of kF on temperature at constant viscosity gave the proper activation parameters, which agree with those obtained in experiments with solutions having a constant viscosity. This agreement validates the latter method, which is more efficient than the former, for determining activation parameters of processes that are modulated by viscosity. The smooth change in kF is governed by the change in viscosity, not in other properties of the solvent, and it does not depend on the choice of the viscosigen. Donor/acceptor electronic coupling (HAB) and reorganizational energy (lambda), obtained by fitting of the temperature dependence of kF to the Marcus equation, are consistent with true electron transfer and with electron transfer that is coupled to, or gated by, a preceding structural rearrangement of the diprotein complex 3Zncyt/pc(II). The fact that at very high viscosity kF approaches zero shows that the reaction is probably gated throughout the investigated range of viscosity. Kinetic effects and noneffects of ionic strength, viscosity, and thermodynamic driving force indicate, but do not prove, that the reaction under consideration is gated. The kinetic effect of viscosity is analyzed in terms of two models. Because ln kF is a nonlinear function of ln eta, protein friction has to be considered in the analysis of viscosity effects on kinetics. PMID- 9201963 TI - Solution conformations of a peptide containing the cytoplasmic domain sequence of the beta amyloid precursor protein. AB - The cytoplasmic domain of the beta amyloid precursor protein (betaAPP) may play a role in cellular events that lead to the secretion of the Abeta peptide, the major constituent of amyloid plaques found in the brains of individuals affected by Alzheimer's disease, by interacting with cellular factors involved in betaAPP function or processing. In order to elucidate the structural basis of cytoplasmic domain activity, the conformations adopted in solution by a peptide containing the 47-residue C-terminal sequence of betaAPP have been investigated by NMR and CD spectroscopy. The peptide does not have a stable tertiary structure, but local regions of the polypeptide chain populate defined conformations. In particular, the amino acid sequences TPEE and NPTY form type I reverse turns. These structured regions correspond to sequences within the cytoplasmic domain implicated in the biological activity of betaAPP. PMID- 9201964 TI - Three-dimensional structure and position of porcine motilin in sodium dodecyl sulfate micelles determined by 1H NMR. AB - The solution structure of the porcine gastrointestinal peptide hormone motilin was determined in the presence of sodium dodecyl sulfate (SDS) micelles at 28 degrees C using 1H nuclear magnetic resonance, full relaxation matrix analysis, and structure calculations based on restrained molecular dynamics. The structure of motilin in SDS micelles is described by a reverse gamma-turn and a beta-turn of type II in the N terminal end, an alpha-helical region in the middle of the molecule, and an extended structure at the C terminus. The position of the motilin molecule relative to the SDS micelles was probed by adding spin-labeled stearic acids, containing 12-doxyl or 5-doxyl spin-labels. We observed selective broadening of the proton resonances of residues 3-5 and concluded that they must be located in the interior of the micelle. These experiments suggest a structural model in which the hydrophobic N terminus consists of two well-defined turns buried in the interior of the micelle, whereas the amphiphilic alpha-helical part is located at the surface of the micelle. Spectral density mapping using a 13C label on the alphaC of Leu10 gave overall rotational correlation times taum of 6.6 and 4.5 ns at 35 and 45 degrees C, respectively. The long correlation time in combination with a high order parameter (S = 0.92) indicates that motilin has a rigid structure in the complex with the SDS micelle. PMID- 9201965 TI - Structures of Cys319 variants and acetohydroxamate-inhibited Klebsiella aerogenes urease. AB - Cys319 is located on a mobile flap covering the active site of Klebsiella aerogenes urease but does not play an essential role in catalysis. Four urease variants altered at position C319 range from having high activity (C319A) to no measurable activity (C319Y), indicating Cys is not required at this position, but its presence is highly influential [Martin, P. R., & Hausinger, R. P. (1992) J. Biol. Chem. 267, 20024-20027]. Here, we present 2.0 A resolution crystal structures of C319A, C319S, C319D, and C319Y proteins and the C319A variant inhibited by acetohydroxamic acid. These structures show changes in the hydration of the active site nickel ions and in the position and flexibility of the active site flap. The C319Y protein exhibits an alternate conformation of the flap, explaining its lack of activity. The changes in hydration and conformation suggest that there are suboptimal protein-solvent and protein-protein interactions in the empty urease active site which contribute to urease catalysis. Specifically, we hypothesize that the suboptimal interactions may provide a significant source of substrate binding energy, and such hidden energy may be a common phenomenon for enzymes that contain mobile active site loops and undergo an induced fit. The acetohydroxamic acid-bound structure reveals a chelate interaction similar to those seen in other metalloenzymes and in a small molecule nickel complex. The inhibitor binding mode supports the proposed mode of urea binding. We complement these structural studies with extended functional studies of C319A urease to show that it has enhanced stability and resistance to inhibition by buffers containing nickel ions. The near wild-type activity and enhanced stability of the C319A variant make it useful for further studies of urease structure-function relationships. PMID- 9201966 TI - Adsorption of pulmonary surfactant protein D to phospholipid monolayers at the air-water interface. AB - The intrinsic surface activity of recombinant rat surfactant-associated protein D (SP-D) expressed in CHO-K1 cells has been determined from measurements of surface tension of its aqueous solutions. The interactions of recombinant SP-D with monolayers of phosphatidylcholine (PC), phosphatidylglycerol (PG), and phosphatidylinositol (PI) spread at the air-water interface have been characterized. Injection of SP-D beneath preformed lipid monolayers at surface pressures less than 30 mN/m produced an increase in surface pressure, consistent with SP-D penetrating the lipid films. The adsorption of SP-D to the lipid monolayers did not display significant head group dependency, suggesting that the changes in surface pressure produced by the protein were likely due primarily to hydrophobic interactions with the lipid layers. In the presence of calcium ions in the subphase, SP-D displayed lower surface activity by itself and a reduced ability to generate surface pressure changes during adsorption to lipid monolayers compared to these properties of the protein in the absence of 2 mM Ca2+. Circular dichroism measurements on SP-D solutions with or without Ca2+ suggested that the cations altered the conformation of the protein and this possibly led to the calcium dependency of the surface activity of the protein in the presence or absence of lipid monolayers. Compressional isotherms of surface pressure versus area for SP-D/(DPPC-PI) and SP-D/(DPPC-PG) films formed by adsorption of the protein to preformed lipid monolayers were consistent with incorporation of some or all of the SP-D molecules into the lipid layers. The isotherms obtained on compression of the SP-D/lipid films to a maximum surface pressure of about 70 mN/m were consistent with the interpretation that any SP-D which was incorporated by adsorption was apparently not squeezed out, nor was lipid removed by the protein. The work suggests that when soluble recombinant SP D is allowed to interact with phospholipid monolayers under the selected conditions of this experiment, it does so to a limited extent driven by primarily hydrophobic forces, and apparently without a high selectivity for phospholipid head groups. PMID- 9201967 TI - Functional interactions between synthetic alkyl phospholipids and the ABC transporters P-glycoprotein, Ste-6, MRP, and Pgh 1. AB - The ABC superfamily of transporters includes the mammalian P-glycoprotein family (Class I and Class II P-gps), the multidrug resistance-associated protein (MRP), the Pgh-1 product of Plasmodium falciparum gene pfmdr1, all of which are associated with cellular pleiotropic drug resistance phenomena. STE6, the yeast transporter for the farnesylated peptide pheromone a, is also a member of this family. Structural similarities in this family translate into functional homology as expression of mouse Mdr3S (P-gp), P. falciparum Pgh-1, and human MRP partially restore mating in a sterile yeast mutant lacking a functional STE6 gene. The demonstration that Class II P-gps function as phosphatidylcholine (PC) translocators raise the possibility that other ABC transporters may also interact with physiological lipids. We report the identification of the synthetic lipid and PC analog ET-18-OCH3 (edelfosine) as a substrate for not only Class II P-gp but also for Class I P-gps and surprisingly for the other ABC transporters MRP, Pgh-1, and STE6. Expression of these proteins in the yeast Saccharomyces cerevisiae JPY201 was found to confer cellular resistance to cytotoxic concentrations of this lipid by a factor of 4-20-fold in a growth inhibition assay. The noted activity of ABC transporters toward this synthetic lipid was specific as a mutant variant of Mdr3 (Mdr3F) with reduced activity could not convey cellular resistance to ET-18-OCH3. ET-18-OCH3 was also found capable of blocking a-peptide pheromone transport and STE6 complementation by these ABC proteins. The inhibitory effect of ET-18-OCH3 on cell growth and a-factor transport could be abrogated by incubation with the lipid acceptor protein BSA or by enzymatic cleavage by microsomal alkylglycerol mono-oxygenase (MAMO). MAMO and BSA reversal of the ether lipid effect was only seen in the presence of a functional transporter. These results suggest that the group of cytotoxic synthetic PC analogs studied reveal possible structural and functional aspects common to the ABC transporters tested. Furthermore, the studies with BSA and MAMO suggest that the mechanism of transport of ET-18-OCH3 by these ABC transporters may be related to the flippase mechanism of PC transport by Mdr2. PMID- 9201968 TI - Annexin V interaction with phosphatidylserine-containing vesicles at low and neutral pH. AB - Annexin V belongs to a class of Ca2+-binding proteins for which different functions in the cell are discussed, e.g., involvement in exocytosis, inhibition of protein kinase C, and calcium channel activity in cartilage matrix vesicles. All these functions are related to the ability of the annexins to bind to acidic phospholipids. In this study, the interaction of annexin V with large unilamellar vesicles (LUV) prepared from phosphatidylserine (PS) at low pH was compared to that at neutral pH. Annexin V strongly binds to PS LUV at low pH, whereas at neutral pH 20-100 microM Ca2+ are required to induce binding. This is caused by the different electric charge of the protein. The binding ability of the PS LUV for annexin V is higher at low pH. Binding of annexin V induces dehydration of the vesicle surface and a decrease of the lateral diffusion within the bilayer. While this dehydration is due to vesicle contact at pH 4, at pH 7.4 it is due to surface covering by annexin V. Annexin V promotes the phospholipid intermixing between LUVs at pH 5 and below but inhibits it at pH 7.4. A substitution of up to 80% of the PS by the uncharged phosphatidylcholine does not impair the extent of phospholipid intermixing at pH 4. The high binding capacity of PS LUV, the disappearance of the inhibiting action, and a calculated increase of the annexin V hydrophobicity make it likely that annexin V is able to penetrate into the membrane at low pH. At neutral pH, annexin V molecules act as steric barriers, preventing close apposition of two vesicles. At pH 5, annexin V lowers the threshold concentration of the Ca2+-induced phospholipid intermixing. Such a promotion is well-known for annexin VII (synexin). The effect may be related to the isoelectric points of the two annexins which have been reported as 4.8 (annexin V) and 7.0 (annexin VII), respectively. PMID- 9201969 TI - Properties and regulation of cytosolic phospholipase A2. PMID- 9201970 TI - Molecular mechanism and functional significance of the MinK control of the KvLQT1 channel activity. AB - The very slowly activating delayed rectifier K+ channel IKs is essential for controlling the repolarization phase of cardiac action potentials and K+ homeostasis in the inner ear. The IKs channel is formed via the assembly of two transmembrane proteins, KvLQT1 and MinK. Mutations in KvLQT1 are associated with a long QT syndrome that causes syncope and sudden death and also with deafness. Here, we show a new mode of association between ion channel forming subunits in that the cytoplasmic C-terminal end of MinK interacts directly with the pore region of KvLQT1. This interaction reduces KvLQT1 channel conductance from 7.6 to 0.58 picosiemens. However, because MinK also reveals a large number of previously silent KvLQT1 channels (x 60), the overall effect is a large increase (x 4) in the macroscopic K+ current. Conformational changes associated with the KvLQT1/MinK association create very slow and complex activation kinetics without much alteration in the deactivation process. Changes induced by MinK have an essential regulatory role in the development of this K+ channel activity upon repetitive electrical stimulation with a particular interest in tachycardia. PMID- 9201971 TI - Human factor IX binds to specific sites on the collagenous domain of collagen IV. AB - The primary region of factor IX that mediates binding to bovine aortic endothelial cells resides in residues 3-11 of the N-terminal region known as the Gla domain. Recently, it was proposed that the observed binding to endothelial cells is actually a measure of the interaction between factor IX and collagen IV (Cheung, W. F., van den Born, J., Kuhn, K., Kjellen, L., Hudson, B. G., and Stafford, D. W. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 11068-11073). To confirm that factor IX binds to collagen IV and to examine the specificity of this interaction, we used scanning force microscopy to examine factor IX binding to collagen IV. We imaged collagen IV in the presence and the absence of factor IX and observed specific interactions between factor IX and collagen IV. Our results demonstrate that factor IX binds to collagen IV at specific sites in the collagenous domain approximately 98 and approximately 50 nm from the C-terminal pepsin-cleaved end. PMID- 9201972 TI - Amino acid residues responsible for galactose recognition in yeast Gal2 transporter. AB - A novel, systematic approach was used to identify amino acid residues responsible for substrate recognition in the transmembrane 10 region of the Gal2 galactose transporter of Saccharomyces cerevisiae. A mixture of approximately 25,000 distinct plasmids that encode all the combinations of 12 amino acids in transmembrane 10 that are different in Gal2 and the homologous glucose transporter Hxt2 was synthesized. Selection of galactose transport-positive clones on galactose limited agar plates yielded 19 clones, all of which contained the Tyr446 residue found in Gal2. 14 of the 19 clones contained Trp455 found in Gal2, whereas the other 5 contained Cys455, a residue not found in either Gal2 or Hxt2. When Tyr446 of Gal2 was replaced with any of the other 19 amino acids, no galactose transport activity was observed in the resulting transporters, indicating that Tyr446 plays an essential role in the transport of this sugar. Replacement of 2 amino acids of Hxt2 with the corresponding Tyr446 and Trp455 of Gal2 allowed the modified Hxt2 to transport galactose. The Km of galactose transport for the modified transporter was 8-fold higher than that of Gal2. These results and other evidence unequivocally show that Tyr446 is essential and Trp455 is important for the discrimination of galactose versus glucose. PMID- 9201973 TI - Activation of c-Jun N-terminal kinase antagonizes an anti-apoptotic action of Bcl 2. AB - Bcl-2 is an intracellular membrane-associated protein that prevents cell death induced by a variety of apoptotic stimuli. A mechanism by which Bcl-2 exerts an anti-cell death effect is, however, not fully understood. In the present study, Bcl-2 suppressed cell death of N18TG neuroglioma cells caused by various apoptotic stresses, including etoposide, staurosporine, anisomycin, and ultraviolet irradiation. Concomitantly, Bcl-2 disrupted a signaling cascade to the c-Jun N-terminal kinase activation induced by the apoptotic stresses. Bcl-2 also prevented the etoposide-induced stimulation of MEKK1. Furthermore, overexpression of c-Jun N-terminal kinase antagonized the death-protective function of Bcl-2. These data suggest that suppression of the c-Jun N-terminal kinase signaling pathway may be critical for Bcl-2 action. PMID- 9201974 TI - Protein phosphatase 2A inhibits nuclear telomerase activity in human breast cancer cells. AB - Most cancer cells have increased levels of telomerase activity implicated in cell immortalization. Activation of telomerase, a ribonucleoprotein complex, catalyzes the elongation of the ends of mammalian chromosomal DNA (telomeres), the length of which regulates cell proliferation. Currently, how telomerase is regulated in cancer is not yet established. The present study shows that telomerase activity is regulated by protein phosphorylation in human breast cancer cells. Incubation of cell nuclear telomerase extracts with protein phosphatase 2A (PP2A) abolished the telomerase activity; in contrast cytoplasmic telomerase activity was unaffected, and protein phosphatases 1 and 2B were ineffective. Inhibition of telomerase activity by PP2A was both concentration- and time-dependent and was prevented by the protein phosphatase inhibitor okadaic acid. In addition, nuclear telomerase inhibited by PP2A was reactivated by endogenous protein kinase(s) in the presence of ATP, but not in the presence of ATPgammaS. Furthermore, telomerase activity in cultured human breast cancer PMC42 cells was stimulated by okadaic acid, consistent with a role for PP2A in the regulation of telomerase activity in intact cells. These findings suggest that protein phosphorylation reversibly regulates the function of telomerase and that PP2A is a telomerase inhibitory factor in the nucleus of human breast cancer cells. PMID- 9201975 TI - Human breast cancer growth inhibited in vivo by a dominant negative pleiotrophin mutant. AB - Pleiotrophin (PTN) is a recently described 18- kDa heparin binding growth/differentiation factor. It also is a proto-oncogene; cells transformed by the Ptn gene form highly angiogenic tumors when implanted into the nude mouse. PTN may be an important regulator of transformation in other tumors, because constitutively high levels of expression of the pleiotrophin (Ptn) gene are found in human breast cancer and other malignant cell lines, and its levels of expression are high in many human tumor specimens. To determine whether PTN is an important regulator of the malignant phenotype of human breast cancer cells, we constructed a mutant cDNA to encode a truncated PTN designed to heterodimerize with the product of the endogenous Ptn gene during processing. The mutant gene product blocked transformation of NIH 3T3 cells by the wild type (wt) Ptn gene product. The mutant Ptn cDNA was then introduced into human breast cancer MDA-MB 231 cells, and clonal lines that stably express the mutant Ptn cDNA were selected. The truncated PTN was shown to form heterodimers with the endogenous Ptn gene product in these cells. Furthermore, the MDA-MB-231 cells that express the mutant Ptn gene were no longer transformed; they failed to form plaques or colonies in soft agar and were unable to form tumors in the athymic nude mouse. The results establish an important role of PTN in the dysregulated growth of human breast cancer cells and suggest that constitutive expression of PTN may be essential to the malignant phenotype of human breast cancers in vivo. PMID- 9201976 TI - Mutants of rat intestinal fatty acid-binding protein illustrate the critical role played by enthalpy-entropy compensation in ligand binding. AB - Site-specific variants of rat intestinal fatty acid-binding protein were constructed to identify the molecular interactions that are important for binding to fatty acids (FAs). Several variants displayed affinities that appeared incompatible with the crystal structure of the protein-FA complex. Thermodynamic measurements provided an explanation for these apparent inconsistencies and revealed that binding affinities often inaccurately reported changes in protein FA interactions because changes in the binding entropy and enthalpy were usually compensatory. These results demonstrate that understanding the effects of amino acid replacements on ligand binding requires measurements of enthalpy and entropy, in addition to affinity. PMID- 9201977 TI - Gene knockout reveals a novel gene cluster for the synthesis of a class of cell wall lipids unique to pathogenic mycobacteria. AB - Surface-exposed unusual lipids containing phthiocerol and phenolphthiocerol are found only in the cell wall of slow-growing pathogenic mycobacteria and are thought to play important roles in host-pathogen interaction. The enzymology and molecular genetics of biosynthesis of phthiocerol and phenolphthiocerol are unknown. We postulate the domain organization of a set of multifunctional enzymes and a cluster of genes (pps) that would encode these enzymes for the biosynthesis of phthiocerol and phenolphthiocerol. A cosmid containing the postulated pps gene cluster was identified by screening a genomic library of Mycobacterium bovis BCG with the postulated homologous domains from mycocerosic acid synthase and fatty acid synthase genes as probes. Homologous cosmids were also identified in the genomic libraries of Mycobacterium tuberculosis and Mycobacterium leprae. M. bovis BCG was transformed with a pps disruption construct, made from the BCG cosmid by introducing the hygromycin resistance gene as the positive-selectable marker and the sacB gene as the counter-selectable marker. Gene disruption by homologous recombination with double crossover was confirmed by polymerase chain reaction and Southern hybridization. Chromatographic analysis showed that the phenolphthiocerol derivative, mycoside B, and phthiocerol dimycocerosates were not produced by the gene knockout mutants. This result confirms the identity of the pps genes. With the identification of the pps gene clusters in both M. tuberculosis and M. leprae, it should be possible to test the postulated roles of these unique lipids in tuberculosis and leprosy. PMID- 9201978 TI - Expression, purification, and properties of recombinant barley (Hordeum sp.) hemoglobin. Optical spectra and reactions with gaseous ligands. AB - A cDNA encoding barley hemoglobin (Hb) has been cloned into pUC 19 and expressed in Escherichia coli. The resulting fusion protein has five extra amino acids at the N terminus compared with the native protein, resulting in a protein of 168 amino acids (18.5 kDa). The recombinant Hb is expressed constitutively. Extracts made from the bacteria containing the recombinant fusion construct contain a protein with a subunit molecular mass of approximately 18.5 kDa comprising approximately 5% total soluble protein. Recombinant Hb was purified to homogeneity according to SDS-polyacrylamide gel electrophoresis by sequential polyethylene glycol precipitation and fast protein liquid chromatography. Its native molecular mass as assessed by fast protein liquid chromatography-size exclusion was 40 kDa suggesting that it is a dimer. Ligand binding experiments demonstrate that 1) barley Hb has a very slow oxygen dissociation rate constant (0.0272 s-1) relative to other Hbs, and 2) the heme of ferrous and ferric forms of the barley Hb is low spin six-coordinate. The subunit structure, optical spectrum, and oxygen dissociation rate of native barley hemoglobin are indistinguishable from those obtained for the recombinant protein. The implications of these kinetic data on the in vivo function of barley Hb are discussed. PMID- 9201979 TI - Differential inhibition of the yeast bc1 complex by phenanthrolines and ferroin. Implications for structure and catalytic mechanism. AB - o-Phenanthroline and m-phenanthroline both inhibit the electron transfer activity of lauryl maltoside-solubilized yeast bc1 complex progressively with time. Pre steady-state kinetics indicate that these compounds bind to the complex on the intermembrane space side, thereby blocking reduction of cytochrome b via the ubiquinol oxidation site. o-Phenanthroline is additionally capable of chelating an iron atom derived from the Rieske Fe-S cluster, thereby distorting the structure of the Rieske protein. EPR analysis shows that the secondary effect of o-phenanthroline occurs after initial inactivation and that m-phenanthroline, which lacks chelating activity, does not affect the Rieske Fe-S cluster. Spectral analysis shows that the b and c1 cytochromes are still dithionite-reducible after inactivation by o-phenanthroline, indicating that they remain intact. Inactivation by o-phenanthroline can be prevented by the addition of Fe2+. Surprisingly, ferroin, the o-phenanthroline-ferrous sulfate complex, also inhibits the bc1 complex activity. In contrast to o-phenanthroline, this effect is instantaneous. The two types of inhibition are clearly distinguishable by pre steady-state reduction kinetics. Interestingly, ferroin can only inhibit electron transfer activity by about 50%. This behavior is discussed in relation to the dimeric structure of the bc1 complex, and we conclude that ferroin binds to only one of the two protomers. The rate of inactivation by o-phenanthroline is dependent on the incubation temperature and can be quantitated in terms of the half-life for a certain temperature, the time at which the bc1 activity is reduced to 50%. In contrast to the solubilized form, the bc1 complex in intact mitochondria is insensitive to o-phenanthroline, suggesting that the inactivation rate by o-phenanthroline is dependent on accessibility of the complex to the agent. Reaction with o-phenanthroline is thus a useful technique for study of structural stability of the bc1 complex under different conditions and should provide a sensitive tool for determination of the relative stability of mutant enzymes. PMID- 9201980 TI - Rapid protein sequencing by tandem mass spectrometry and cDNA cloning of p20 CGGBP. A novel protein that binds to the unstable triplet repeat 5'-d(CGG)n-3' in the human FMR1 gene. AB - The autonomous expansion of the unstable 5'-d(CGG)n-3' repeat in the 5' untranslated region of the human FMR1 gene leads to the fragile X syndrome, one of the most frequent causes of mental retardation in human males. We have recently described the isolation of a protein p20-CGGBP that binds sequence specifically to the double-stranded trinucleotide repeat 5'-d(CGG)-3' (Deissler, H., Behn-Krappa, A., and Doerfler, W. (1996) J. Biol. Chem. 271, 4327-4334). We demonstrate now that the p20-CGGBP can also bind to an interrupted repeat sequence. Peptide sequence tags of p20-CGGBP obtained by nanoelectrospray mass spectrometry were screened against an expressed sequence tag data base, retrieving a clone that contained the full-length coding sequence for p20-CGGBP. A bacterially expressed fusion protein p20-CGGBP-6xHis exhibits a binding pattern to the double-stranded 5'-d(CGG)n-3' repeat similar to that of the authentic p20 CGGBP. This novel protein lacks any overall homology to other known proteins but carries a putative nuclear localization signal. The p20-CGGBP gene is conserved among mammals but shows no homology to non-vertebrate species. The gene encoding the sequence for the new protein has been mapped to human chromosome 3. PMID- 9201981 TI - Evidence for a unique mechanism of strand transfer from the transactivation response region of HIV-1. AB - We previously found that strand transfer by human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) is promoted at sites where RT pauses during synthesis. In this report, strand transfer is measured within the 5' transactivation response region (TAR) of HIV-1 RNA. We hypothesized that the stable hairpin structure of TAR would induce RT pausing, promoting RNase H directed cleavage of the template and subsequent transfer at that site. We further predicted that HIV-1 nucleocapsid protein (NC), known to melt secondary structures, would decrease transfer. We show that TAR created a strong pause site for RT, but NC significantly promoted strand transfer. The effect of NC is specific, since other single strand binding proteins failed to stimulate transfer. In another unexpected outcome, preferred positions of internal transfer were not at the pause site but were in the upper stem and loop of TAR. Thus, we propose a new mechanism for transfer within TAR described by an interactive hairpin model, in which association between the donor and the acceptor templates within the TAR stem promotes transfer. The model is consistent with the observed stimulation of strand transfer by NC. The model is applicable to internal and replicative end transfer. PMID- 9201982 TI - Cloning and characterization of HB2, a candidate high density lipoprotein receptor. Sequence homology with members of the immunoglobulin superfamily of membrane proteins. AB - The protection against coronary artery disease attributed to high density lipoprotein (HDL) may be associated with several functions, including its central role in reverse cholesterol transport, possible antioxidant and antithrombotic properties and others not yet identified which may depend on specific interactions between HDL and cell receptors. Several HDL-binding proteins have been identified including two candidate liver HDL receptors, HB1 and HB2 recently purified in this laboratory. We now report the cloning, sequencing, and some properties of HB2, the most abundant of the pair. It shows significant homology with the adhesion molecules ALCAM and BEN of the immunoglobulin superfamily and the cDNA, when transfected into HepG2 or COS cells, caused specific HDL3 binding to increase by 80-100%. Further, ligand blotting of glycoproteins isolated from phorbol 12-myristate 13-acetate-treated THP-1 cells or from transfected HepG2 and Chinese hamster ovary cells also provided evidence of increased binding of HDL3 to HB2. Differentiation of THP-1 cells into macrophages resulted in a striking increase in HB2 mRNA which was attenuated if cells were cholesterol-loaded by incubation with acetylated low density lipoprotein. If the interaction between HDL and HB2 reduces the adhesion-induced inflammatory cellular events that characterize arterial wall injury, thereby achieving the protection associated with higher plasma levels of HDL, these findings may provide a clue to one mitigating effect of HDL in heart disease. PMID- 9201983 TI - Flexible DNA: genetically unstable CTG.CAG and CGG.CCG from human hereditary neuromuscular disease genes. AB - The properties of duplex CTG.CAG and CGG.CCG, which are involved in the etiology of several hereditary neurodegenerative diseases, were investigated by a variety of methods, including circularization kinetics, apparent helical repeat determination, and polyacrylamide gel electrophoresis. The bending moduli were 1.13 x 10(-19) erg.cm for CTG and 1.27 x 10(-19) erg.cm for CGG, approximately 40% less than for random B-DNA. Also, the persistence lengths of the triplet repeat sequences were approximately 60% the value for random B-DNA. However, the torsional moduli and the helical repeats were 2.3 x 10(-19) erg.cm and 10.4 base pairs (bp)/turn for CTG and 2.4 x 10(-19) erg.cm and 10.3 bp/turn for CGG, respectively, all within the range for random B-DNA. Determination of the apparent helical repeat by the band shift assay indicated that the writhe of the repeats was different from that of random B-DNA. In addition, molecules of 224 245 bp in length (64-71 triplet repeats) were able to form topological isomers upon cyclization. The low bending moduli are consistent with predictions from crystallographic variations in slide, roll, and tilt. No unpaired bases or non-B DNA structures could be detected by chemical and enzymatic probe analyses, two dimensional agarose gel electrophoresis, and immunological studies. Hence, CTG and CGG are more flexible and highly writhed than random B-DNA and thus would be expected to act as sinks for the accumulation of superhelical density. PMID- 9201984 TI - Triplet repeat instability and DNA topology: an expansion model based on statistical mechanics. AB - The variance of writhe, the contribution of writhe to supercoiling, and the free energies of supercoiling were calculated for (CTG.CAG)n and (CGG.CCG)n triplet repeat sequences (TRS) by statistical mechanics from the bending and torsional moduli previously determined. Expansions of these sequences are inherited by non mendelian transmission and are linked with several hereditary neuromuscular diseases. The variance of writhe was greater for the TRS than for random B-DNA. For random B-DNA, (CGG)n, and (CTG)n, the contribution of writhe to supercoiling was 70, 78, and 79%, whereas the free energy of supercoiling at a length of 10 kilobase pairs was 1040.RT, 760.RT, and 685.RT, respectively. These data indicate that the TRS are preferential sites for the partitioning of supercoiling. Calculations of the differences in free energy of supercoiling between the TRS and random B-DNA revealed a local minimum at approximately 520 base pairs. Human medical genetic studies have shown that individuals carrying up to 180-200 copies of TRS (540-600 base pairs, premutations) in the fragile X or myotonic dystrophy gene loci are usually asymptomatic, whereas large expansions (>200 repeats, full mutations), which lead to disease, are observed in their offspring. Therefore, the length corresponding to the local minimum in free energy of supercoiling correlates with the genetic breakpoint between premutation and full mutation. We propose that (a) TRS instability is mediated by DNA mispairing caused by the accumulation of supercoiling within the repeats, and (b) the expansions that take place at the premutation to full mutation threshold are associated with increased mispairing caused by the optimal partitioning of writhe within the TRS at this length. PMID- 9201985 TI - Hairpin formation during DNA synthesis primer realignment in vitro in triplet repeat sequences from human hereditary disease genes. AB - Genetic expansion of DNA triplet repeat sequences (TRS) found in neurogenetic disorders may be due to abnormal DNA replication. We have previously observed strong DNA synthesis pausings at specific loci within the long tracts (> approximately 70 repeats) of CTG.CAG and CGG.CCG as well as GTC.GAC by primer extensions in vitro using DNA polymerases (the Klenow fragment of Escherichia coli DNA polymerase I, the modified T7 DNA polymerase (Sequenase), and human DNA polymerase beta). Herein, we have isolated and analyzed the products of stalled synthesis found at approximately 30-40 triplets from the beginning of the TRS. DNA sequence analyses revealed that the stalled products contained short tracts of homogeneous TRS (6-12 repeats) in the middle of the sequence corresponding to the flanking region of the primer-template sequence. The sequence at the 3'-side terminated at the end of the primer, indicating that the primer molecule had served as a template. In addition, chemical probe and polyacrylamide gel electrophoretic analyses revealed that the stalled products existed in hairpin structures. We postulate that these products of the DNA polymerases are caused by the existence of an unusual DNA conformation(s) within the TRS, during the in vitro DNA synthesis, enhancing the DNA slippages and the hairpin formations in the TRS due to primer realignment. The consequence of these steps is DNA synthesis to the end of the primer and termination. Primer realignment including hairpin formation may play an important intermediate role in the replication of TRS in vivo to elicit genetic expansions. PMID- 9201986 TI - Studies on the symmetry and sequence context dependence of the HIV-1 proteinase specificity. AB - Two major types of cleavage sites with different sequence preferences have been proposed for the human immunodeficiency virus type 1 (HIV-1) proteinase. To understand the nature of these sequence preferences better, single and multiple amino acid substitutions were introduced into a type 1 cleavage site peptide, thus changing it to a naturally occurring type 2 cleavage site sequence. Our results indicated that the previous classification of the retroviral cleavage sites may not be generally valid and that the preference for a residue at a particular position in the substrate depends strongly on the neighboring residues, including both those at the same side and at the opposite side of the peptide backbone of the substrate. Based on these results, pseudosymmetric (palindromic) substrates were designed. The retroviral proteinases are symmetrical dimers of two identical subunits; however, the residues of naturally occurring cleavage sites do not show symmetrical arrangements, and no obvious symmetrical substrate preference has been observed for the specificity of HIV proteinase. To examine the role of the asymmetry created by the peptide bonds on the specificity of the respective primed and nonprimed halves of the binding site, amino acid substitutions were introduced into a palindromic sequence. In general, the results suggested that the asymmetry does not result in substantial differences in specificity of the S3 and S3' subsites, whereas its effect is more pronounced for the S2 and S2' subsites. Although it was possible to design several good palindromic substrates, asymmetrical arrangements may be preferred by the HIV proteinase. PMID- 9201987 TI - Interactions between the human RNA polymerase II subunits. AB - As an initial approach to characterizing the molecular structure of the human RNA polymerase II (hRPB), we systematically investigated the protein-protein contacts that the subunits of this enzyme may establish with each other. To this end, we applied a glutathione S-transferase-pulldown assay to extracts from Sf9 insect cells, which were coinfected with all possible combinations of recombinant baculoviruses expressing hRPB subunits, either as untagged polypeptides or as glutathione S-transferase fusion proteins. This is the first comprehensive study of interactions between eukaryotic RNA polymerase subunits; among the 116 combinations of hRPB subunits tested, 56 showed significant to strong interactions, whereas 60 were negative. Within the intricate network of interactions, subunits hRPB3 and hRPB5 play a central role in polymerase organization. These subunits, which are able to homodimerize and to interact, may constitute the nucleation center for polymerase assembly, by providing a large interface to most of the other subunits. PMID- 9201988 TI - Characterization of beta-1,2-mannosyltransferase in Candida guilliermondii and its utilization in the synthesis of novel oligosaccharides. AB - A particulate insoluble enzyme fraction containing mannosyltransferases from Candida guilliermondii IFO 10279 strain cells was obtained as the residue after extracting a 105,000 x g pellet of cell homogenate with 1% Triton X-100. Incubation of this fraction with a mannopentaose, Manalpha1-->3(Manalpha1- >6)Manalpha1-->2Manalpha1+ ++-->2Man, in the presence of GDP-mannose and Mn2+ ion at pH 6.0 gave a third type of beta-1,2 linkage-containing mannohexaose, Manbeta1 ->2Manalpha1-->3(Manalpha1-->6)Manalpha1++ +-->2Manalpha1-->2Man , the structure of which was identified by means of a sequential NMR assignment. The results of a substrate specificity study indicated that the beta-1,2-mannosyltransferase requires a mannobiosyl unit, Manalpha1--> 3Manalpha1-->, at the nonreducing terminal site. We synthesized novel oligosaccharides using substrates possessing a nonreducing terminal alpha-1,3-linked mannose unit prepared from various yeast mannans. Further incubation of the enzymatically synthesized oligosaccharide with the enzyme fraction gave the following structure, Manbeta1-->2Manbeta1- >2Manalpha1-->3(Manalpha1- ->6)Manalpha1--> 2Manalpha1-->2Man, which has been found to correspond to antigenic factor 9. Incubation of Candida albicans serotype B mannan with the enzyme fraction gave significantly transformed mannan, which contains the third type of beta-1,2-linked mannose units. PMID- 9201989 TI - Fluorescence probing of yeast actin subdomain 3/4 hydrophobic loop 262-274. Actin actin and actin-myosin interactions in actin filaments. AB - Residues 262-274 form a loop between subdomains 3 and 4 of actin. This loop may play an important role in actin filament formation and stabilization. To assess directly the behavior of this loop, we mutated Ser265 of yeast actin to cysteine (S265C) and created another mutant (S265C/C374A) by changing Cys374 of S265C actin to alanine. These changes allowed us to attach a pyrene maleimide stoichiometrically to either Cys374 or Cys265. These mutations had no detectable effects on the protease susceptibility, intrinsic ATPase activity, and thermal stability of labeled or unlabeled G-actin. The presence of the loop cysteine, either labeled or unlabeled, did not affect the actin-activated S1 ATPase activity or the in vitro motility of the actin. Both mutant actins, either labeled or unlabeled, nucleated filament formation considerably faster than wild type (WT) actin, although the critical concentration was not affected. Whereas the fluorescence of the C-terminal (WT) probe increased during polymerization, that of the loop (S265C/C374A) probe decreased, and the fluorescence of the doubly labeled actin (S265C) was approximately 50% less than the sum of the fluorescence of the individual fluorophores. Quenching was also observed in copolymers of labeled WT and S265C/C374A actins. An excimer peak was present in the emission spectrum of labeled S265C F-actin and in the labeled S265C/C374A-WT actin copolymers. These results show that in the filaments, the C-terminal pyrene of a substantial fraction of monomers directly interacts with the loop pyrene of neighboring monomers, bringing the two cysteine sulfurs to within 18 A of one another. Finally, when bound to labeled S265C/C374A F-actin, myosin S1, but not tropomyosin, caused an increase in fluorescence of the loop probe. Both proteins had no effect on excimer fluorescence. These results help establish the orientation of monomers in F-actin and show that the binding of S1 to actin subdomains 1 and 2 affects the environment of the loop between subdomains 3 and 4. PMID- 9201990 TI - Role of heparan sulfate proteoglycans in the uptake and degradation of tissue factor pathway inhibitor-coagulation factor Xa complexes. AB - Tissue factor pathway inhibitor (TFPI) is a potent inhibitor of blood coagulation factor Xa (fXa) and factor VIIa. We have recently shown that fXa binding stimulates the uptake and degradation of cell surface-bound 125I-TFPI (Ho, G., Toomey, J. R., Broze, G. J., Jr., and Schwartz, A. L. (1996) J. Biol. Chem. 271, 9497-9502). In the present study we examined the role of cell surface glycosaminoglycans (GAGs) in this process. Removal of cell surface GAG chains by treatment of cells with heparinase or heparitinase but not chondroitinase markedly reduced fXa-stimulated 125I-TFPI uptake and degradation. Inhibition of GAG sulfation by growth of cells in chlorate-containing medium similarly decreased fXa-stimulated 125I-TFPI degradation. These results suggest that heparan sulfate proteoglycans (HSPGs) are required for the uptake and degradation of 125I-TFPI.fXa complexes. Chemical cross-linking/immunoprecipitation analyses revealed that 125I-TFPI was directly associated with HSPGs on the cell surface and that fXa binding increased the amount of 125I-TFPI bound. Of the several cell lines evaluated, bend endothelial cells demonstrated the greatest fXa stimulation of 125I-TFPI uptake and degradation. Cross-linking/immunoprecipitation analyses on bend cells also revealed that HSPGs were specifically associated with TFPI and fXa. These data suggest that HSPGs may directly act as the uptake and degradation receptor for TFPI.fXa complexes. PMID- 9201991 TI - A novel proximal element mediates the regulation of mouse Ren-1C promoter by retinoblastoma protein in cultured cells. AB - The protein product of the retinoblastoma susceptibility gene, RB, is a nuclear phosphoprotein that modulates transcription of genes involved in growth control via interactions with transcription factors. Renin is a rate-limiting enzyme of the renin-angiotensin system that regulates blood pressure and water-electrolyte balance. Renin gene expression is regulated in a tissue-specific and developmentally linked manner. Similarly, the expression of RB is controlled in a differentiation-linked manner. Thus, to investigate whether RB is involved in the regulation of renin gene expression, we examined the effects of RB on transcriptional activity of the mouse renin (Ren-1C) promoter. The Ren-1C promoter contains two transcriptionally important elements; the RU-1 (-224 to 138) and RP-2 (-75 to -47) elements. RB activated the Ren-1C promoter in human embryonic kidney cells. The promoter element responsible for RB-mediated transcriptional regulation was the RP-2 element. The results of DNA-protein binding experiments showed that RB increased nuclear binding activity to the RP-2 element, and site-directed mutation which disrupted binding of nuclear factors to the RP-2 element markedly reduced RB-mediated activation of Ren-1C promoter in human embryonic kidney cells. These results indicate that the RP-2 element plays an important role in RB-mediated transcriptional regulation of Ren-1C promoter activity in human embryonic kidney cells, thereby suggesting an interesting mechanism by which RB may modulate the renin-angiotensin system. PMID- 9201992 TI - Refolding intermediates of acid-unfolded mitochondrial aspartate aminotransferase bind to hsp70. AB - The cytosolic (cAAT) and mitochondrial (mAAT) isozymes of eukaryotic aspartate aminotransferase share a high degree of sequence identity and almost identical three-dimensional structure. The rat liver proteins can be refolded and reassembled into active dimers after unfolding at low pH. However, refolding of the mitochondrial form after unfolding at pH 2.0 is arrested in the presence of hsp70, whereas this chaperone does not affect the refolding of the cytosolic isozyme unfolded under similar conditions. To elucidate the nature of the differential interaction between hsp70 and the two transaminase forms, we have characterized their refolding from their acid-unfolded states. The recovery of activity of the cytosolic enzyme is monophasic and can be adequately described by a single first-order reaction. By contrast, two sequential first-order rate limiting steps can be detected for the refolding and reactivation of the mitochondrial protein. The overall refolding pathway of mAAT includes a very fast collapse to an intermediate with 80% of the secondary structure of the active dimer. This is followed by a slow isomerization to form assembly-competent monomers that rapidly associate to form an inactive dimer and a final structural rearrangement of the dimer to the native conformation. Analysis of the interaction of hsp70 with intermediates along the folding pathway of mAAT shows that the polypeptide loses its ability to bind to the chaperone after it has proceeded through the first isomerization/fast dimerization steps. Thus it appears that only the first collapsed intermediate states in the folding of mAAT bind hsp70. By contrast a faster refolding of cAAT from this collapsed state could explain, at least in part, the inability of hsp70 to bind this isozyme. PMID- 9201993 TI - Effect of saposins A and C on the enzymatic hydrolysis of liposomal glucosylceramide. AB - The degradation of glucosylceramide in lysosomes is accomplished by glucosylceramidase with the assistance of, at least, another protein, saposin C (Sap C), which is generated from a large precursor together with three other similar proteins, saposins A, B, and D. In the present study, we have examined the effects of saposins on the enzymatic hydrolysis of glucosylceramide inserted in large and small phospholipid liposomes. The glucosylceramide contained in large unilamellar vesicles (LUV) was degraded by glucosylceramidase at a rate 7-8 fold lower than glucosylceramide inserted in small unilamellar vesicles (SUV). The separate addition of either Sap A or Sap C to the LUV system partially stimulated the sphingolipid degradation while saposins B and D had no effect. In the presence of both Sap A and Sap C, the rate of sphingolipid degradation was higher than the sum of the rates with the two saposins individually, indicating synergism in their actions. The stimulatory effect of the two saposins depended on the incorporation of an acidic phospholipid such as phosphatidylserine (PS) into LUV. The characteristics of glucosylceramidase activation by Sap C were different from those of Sap A. Sap C increased the rate of hydrolysis of both the artificial water soluble substrate, 4-methylumbelliferyl-beta-D-glucopyranoside, and the lipid substrate, glucosylceramide, while Sap A only stimulated degradation of the sphingolipid. Also the binding properties of Saps A and C were markedly different. At acidic pH values, Sap C bound to PS-containing LUV and promoted the association of glucosylceramidase with the membrane. In contrast, Sap A had poor affinity for the membrane even in the presence of glucosylceramide; moreover, Sap A did not potentiate the capacity of Sap C to mediate glucosylceramidase binding. In conclusion, our results show that both Sap A and Sap C are required for maximal hydrolysis of glucosylceramide inserted in PS-containing LUV, that their effects are synergistic, and that their mode of action is different. Sap C is responsible for the membrane binding of glucosylceramidase, while Sap A stimulation is possibly related to its effect on the conformation of the enzyme. It can be envisaged that Sap A in conjunction with Sap C might have a physiological role in glucosylceramide degradation. PMID- 9201994 TI - Reconstitution of FhuA, an Escherichia coli outer membrane protein, into liposomes. Binding of phage T5 to Fhua triggers the transfer of DNA into the proteoliposomes. AB - The Escherichia coli outer membrane protein FhuA catalyzes the transport of ferrichrome and is the receptor of bacteriophage T5. Purified FhuA was reconstituted into liposomes. The size of the proteoliposomes and the distribution of the proteins in the vesicles were determined by freeze fracture electron microscopy. Unilamellar vesicles with a diameter larger than 200 nm were observed frequently. FhuA was symetrically oriented in the proteoliposomes. Reconstituted FhuA was functional as binding of phage T5 induced the release of phage DNA and its transfer inside the vesicles. PMID- 9201995 TI - Genomic organization of the 3' region of the human mucin gene MUC5B. AB - MUC5B, mapped clustered with MUC6, MUC2, and MUC5AC to chromosome 11p15.5, is a human mucin gene of which the genomic organization is being elucidated. We have recently published the sequence and the peptide organization of its huge central exon, 10,713 base pairs (bp) in length. We present here the genomic organization of its 3' region, which encompasses 10,690 bp. The genomic sequence has been completely determined. The 3' region of MUC5B is composed of 18 exons ranging in size from 32 to 781 bp, contrasting thus with the very large central exon. The sizes of the 18 introns range from 114 to 1118 bp. Some repetitive sequences were identified in four introns. The peptide deduced from the sequence of the 18 exons consists of an 808-amino acid peptide. This carboxyl-terminal region exhibits extensive sequence similarity to MUC2, MUC5AC, and von Willebrand factor, particularly the number and the positions of the cysteine residues, suggesting that this domain may be derived from a common ancestral gene. The presence in these components of a cystine knot also found in growth factors such as transforming growth factor-beta is of particular interest. Moreover, one part of this peptide is identical to the 196-amino acid sequence deduced from the cDNA clone pSM2-1, which codes for a part of the high molecular weight mucin MG1 isolated from human sublingual gland. Considering the expression pattern of MUC5B and the origin of MG1, we can thus conclude that MUC5B encodes MG1. PMID- 9201996 TI - Discovery of the shortest sequence motif for high level mucin-type O glycosylation. AB - The consensus primary amino acid sequence for mucin-type O-glycosylation sites has not been identified. To determine the shortest motif sequence required for high level mucin-type O-glycosylation, we prepared more than 100 synthetic peptides and assayed in vitro O-GalNAc transfer to serine or threonine in these peptides using a bovine colostrum UDP-N-acetylgalactosamine:polypeptide N acetylgalactosaminyl transferase (O-GalNAcT). We chose the sequence PDAASAAP from human erythropoietin (hEPO) for further systematic substitutions because it accepted GalNAc and was a fairly simple sequence consisting only of four kinds of amino acids. Several substitutions showed that threonine is approximately 40-fold better than serine as the glycosylated amino acid and a proline at position +3 on the C-terminal side is very important. To define the effect of proline residues around the glycosylation site, we analyzed a series of peptides containing one to three proline residues in a parent peptide AAATAAA. The results clearly indicated that prolines at positions +1 and +3 had a positive effect. The O-GalNAc transfer level of AAATPAP was increased approximately 90-fold from AAATAAA. The deletion of amino acids from the N-terminal side of the glycosylation site suggested that five amino acids from position -1 to +3 were especially important for glycosylation. Moreover, the influence of all 20 amino acids at positions -1, +2, and +4 was analyzed. Uncharged amino acids were preferred at position -1, and small or positively charged amino acids were preferred at position +2. No preference was observed at position +4. We propose a mucin-type O-glycosylation motif, XTPXP, which may be suitable as a signal for protein O-glycosylation. The features observed in this study also appear to be very useful for prediction of mucin-type O-glycosylation sites in glycoproteins. PMID- 9201997 TI - Binding specificities of the sialoadhesin family of I-type lectins. Sialic acid linkage and substructure requirements for binding of myelin-associated glycoprotein, Schwann cell myelin protein, and sialoadhesin. AB - The carbohydrate binding specificities of three sialoadhesins, a subgroup of I type lectins (immunoglobulin superfamily lectins), were compared by measuring lectin-transfected COS cell adhesion to natural and synthetic gangliosides. The neural sialoadhesins, myelin-associated glycoprotein (MAG) and Schwann cell myelin protein (SMP), had similar and stringent binding specificities. Each required an alpha2,3-linked sialic acid on the terminal galactose of a neutral saccharide core, and they shared the following rank-order potency of binding: GQ1balpha >> GD1a = GT1b >> GM3 = GM4 >> GM1, GD1b, GD3, GQ1b (nonbinders). In contrast, sialoadhesin had less exacting specificity, binding to gangliosides that bear either terminal alpha2,3- or alpha2,8-linked sialic acids with the following rank-order potency of binding: GQ1balpha > GD1a = GD1b = GT1b = GM3 = GM4 > GD3 = GQ1b >> GM1 (nonbinder). CD22 did not bind to any ganglioside tested. Binding of MAG, SMP, and sialoadhesin was abrogated by chemical modification of either the sialic acid carboxylic acid group or glycerol side chain on a target ganglioside. Synthetic ganglioside GM3 derivatives further distinguished lectin binding specificities. Deoxy and/or methoxy derivatives of the 4-, 7-, 8-, or 9 position of sialic acid attenuated or eliminated binding of MAG, as did replacement of the sialic acid acetamido group with a hydroxyl. In contrast, the 4- and 7-deoxysialic acid derivatives supported sialoadhesin binding at near control levels (the other derivatives did not support binding). These data are consistent with sialoadhesin binding to one face of the sialic acid moiety, whereas MAG (and SMP) may have more complex binding sites or may bind sialic acids only in the context of more restricted oligosaccharide conformations. PMID- 9201998 TI - Functional and structural relationship between the calmodulin-binding, actin binding, and actomyosin-ATPase inhibitory domains on the C terminus of smooth muscle caldesmon. AB - Multiple functional domains responsible for calmodulin (CaM) binding and actin binding/actomyosin ATPase inhibition are present in the region between residues 598-756 of the chicken gizzard smooth muscle caldesmon (CaD) molecule. To precisely localize these functional domains and to further elucidate the structural basis of these domains, we analyzed a series of purified mutants of chicken gizzard smooth muscle CaD generated by internal deletions of amino acid sequences and expression in a baculovirus expression system. Our results demonstrate that, in addition to a strong actin-binding site sequence between residues 718-723 (Wang, Z., and Chacko, S. (1996) J. Biol. Chem. 271, 25707 25714), two weak actin-binding motifs are present in the regions between residues 690-699 and 650-666. These weak actin-binding regions function independently and are associated with weak actomyosin inhibitory activity. Analysis of the CaM binding sites A (residues 658-666) and B (residues 690-695), the major CaM binding sites in the C-terminal region of CaD, provided direct evidence for the involvement of both CaM-binding sites in the CaM-mediated reversal of the inhibition of actomyosin ATPase activity by CaD and for the functional independence of the two CaM-binding sites. Furthermore, the sequences between residues 598-649, upstream of CaM-binding site A, and 700-717, downstream of CaM binding site B, appear to have no effect on either actin-binding or CaM-binding. The data also suggest that both CaM-binding sites A and B structurally overlap or lie in close proximity to the adjacent weak actin-binding sites and weak actomyosin ATPase inhibitory determinants. PMID- 9201999 TI - Dictyostelium myosin heavy chain kinase A subdomains. Coiled-coil and wd repeat roles in oligomerization and substrate targeting. AB - Myosin heavy chain kinase A (MHCK A) participates in the regulation of cytoskeletal myosin assembly in Dictyostelium, driving filament disassembly via phosphorylation of sites in the myosin tail. MHCK A contains an amino-terminal coiled-coil domain, a novel central catalytic domain, and a carboxyl-terminal domain containing a 7-fold WD repeat motif. We have overexpressed MHCK A truncation constructs to clarify the roles of each of these domains. Recombinant full-length MHCK A, MHCK A lacking the predicted coiled-coil domain, and MHCK A lacking the WD repeat domain were expressed at high levels in Dictyostelium cells lacking endogenous MHCK A. Biochemical analysis of the purified proteins demonstrates that the putative coiled-coil domain is responsible for the oligomerization of the MHCK A holoenzyme. Removal of the WD repeat domain had no effect on catalytic activity toward a synthetic peptide, but did result in a 95% loss of protein kinase activity when native myosin filaments were used as the substrate. Cellular analysis confirms that the same severe loss of activity against myosin occurs in vivo when the WD repeat domain is eliminated. These results suggest that the WD repeat domain of MHCK A serves to target this enzyme to its physiological substrate. PMID- 9202000 TI - Relationship of conserved residues in the IMP binding site to substrate recognition and catalysis in Escherichia coli adenylosuccinate synthetase. AB - Gln34, Gln224, Leu228, and Ser240 are conserved residues in the vicinity of bound IMP in the crystal structure of Escherichia coli adenylosuccinate synthetase. Directed mutations were carried out, and wild-type and mutant enzymes were purified to homogeneity. Circular dichroism spectroscopy indicated no difference in secondary structure between the mutants and the wild-type enzyme in the absence of substrates. Mutants L228A and S240A exhibited modest changes in their initial rate kinetics relative to the wild-type enzyme, suggesting that neither Leu228 nor Ser240 play essential roles in substrate binding or catalysis. The mutants Q224M and Q224E exhibited no significant change in KmGTP and KmASP and modest changes in KmIMP relative to the wild-type enzyme. However, kcat decreased 13-fold for the Q224M mutant and 10(4)-fold for the Q224E mutant relative to the wild-type enzyme. Furthermore, the Q224E mutant showed an optimum pH at 6.2, which is 1.5 pH units lower than that of the wild-type enzyme. Tryptophan emission fluorescence spectra of Q224M, Q224E, and wild-type proteins under denaturing conditions indicate comparable stabilities. Mutant Q34E exhibits a 60 fold decrease in kcat compared with that of the wild-type enzyme, which is attributed to the disruption of the Gln34 to Gln224 hydrogen bond observed in crystal structures. Presented here is a mechanism for the synthetase, whereby Gln224 works in concert with Asp13 to stabilize the 6-oxyanion of IMP. PMID- 9202001 TI - Conditional expression of the mitogen-activated protein kinase (MAPK) phosphatase MKP-1 preferentially inhibits p38 MAPK and stress-activated protein kinase in U937 cells. AB - Phorbol ester tumor promoters, such as phorbol 12-myristate 13-acetate (PMA), are potent activators of extracellular signal-regulated kinase 2 (ERK2), stress activated protein kinase (SAPK), and p38 mitogen-activated protein kinase (MAPK) in U937 human leukemic cells. These kinases are regulated by the reversible dual phosphorylation of conserved threonine and tyrosine residues. The dual specificity protein phosphatase MAPK phosphatase-1 (MKP-1) has been shown to dephosphorylate and inactivate ERK2, SAPK, and p38 MAPK in transient transfection studies. Here we demonstrate that PMA treatment induces MKP-1 protein expression in U937 cells, which is detectable within 30 min with maximal levels attained after 4 h. This time course coincides with the rapid inactivation of PMA-induced SAPK activity, but not ERK2 phosphorylation, which remains elevated for up to 6 h. To examine directly the role of MKP-1 in the regulation of these protein kinases in vivo, we established a U937 cell line that conditionally expresses MKP 1 from the human metallothionein IIa promoter. Conditional expression of MKP-1 inhibited PMA-induced ERK2, SAPK, and p38 MAPK activity. By titrating the levels of MKP-1 expression from the human metallothionein IIa promoter, however, it was found that p38 MAPK and SAPK were much more sensitive to inhibition by MKP-1 than ERK2. This differential substrate specificity of MKP-1 can be functionally extended to nuclear transcriptional events in that PMA-induced c-Jun transcriptional activity was more sensitive to inhibition by MKP-1 than either Elk-1 or c-Myc. Conditional expression of MKP-1 also abolished the induction of endogenous MKP-1 protein expression in response to PMA treatment. This negative feedback regulatory mechanism is likely due to MKP-1-mediated inhibition of ERK2, as studies utilizing the MEK1/2 inhibitor PD98059 suggest that ERK2 activation is required for PMA-induced MKP-1 expression. These findings suggest that ERK2 mediated induction of MKP-1 may play an important role in preferentially attenuating signaling through the p38 MAPK and SAPK signal transduction pathways. PMID- 9202002 TI - Kinetics and thioredoxin specificity of thiol modulation of the chloroplast H+ ATPase. AB - The kinetics of thiol modulation of the chloroplast H+-ATPase (CF0CF1) in membrana were analyzed by employing thioredoxins that were kept reduced by 0.1 mM dithiothreitol. The kinetics of thiol modulation depend on the extent of the proton gradient. The process is an exponential function of the thioredoxin concentration and reaction time and can be described by an irreversible second order reaction. The results indicate that the formation of the complex between thioredoxin and CF0CF1 is slow compared with the subsequent reduction step. Furthermore we have compared the efficiencies of the Escherichia coli thioredoxin Trx and the two chloroplast thioredoxins Tr-m and Tr-f. The second order rate constants are 0.057 (Tr-f), 0.024 (Trx), and 0.010 s-1 microM-1 (Tr-m) suggesting that Tr-f rather than Tr-m is the physiological reductant for the chloroplast ATPase. The often employed artificial reductant dithiothreitol exhibits a second order rate constant in thiol modulation of 1.02.10(-6) s-1 microM-1. PMID- 9202003 TI - Ubiquinol:cytochrome c oxidoreductase. The redox reactions of the bis-heme cytochrome b in unenergized and energized submitochondrial particles. AB - The redox reactions of the bis-heme cytochrome b of the ubiquinol:cytochrome c oxidoreductase complex (complex III, bc1 complex) were studied in bovine heart submitochondrial particles (SMP). It was shown that (i) when SMP were treated with the complex III inhibitor myxothiazol (or MOA-stilbene or stigmatellin) or with KCN and ascorbate to reduce the high potential centers of complex III (iron sulfur protein and cytochromes c + c1), NADH or succinate reduced heme bL slowly and incompletely. In contrast, heme bH was reduced by these substrates completely and much more rapidly. Only when the complex III inhibitor was antimycin, and the high potential centers were in the oxidized state, NADH or succinate was able to reduce both bH and bL rapidly and completely. (ii) When NADH or succinate was added to SMP inhibited at complex III by antimycin and energized by ATP, the bis heme cytochrome b was reduced only partially. Prereduction of the high potential centers was not necessary for this partial b reduction, but slowed down the reduction rate. Deenergization of SMP by uncoupling (or addition of oligomycin to inhibit ATP hydrolysis) resulted in further b reduction. Addition of ATP after b was reduced by substrate resulted in partial b oxidation, and the heme remaining reduced appeared to be mainly bL. Other experiments suggested that the redox changes of cytochrome b effected by energization and deenergization of SMP occurred via electronic communication with the ubiquinone pool. These results have been discussed in relation to current concepts regarding the mechanism of electron transfer by complex III. PMID- 9202004 TI - Activating transcription factor 2 (ATF2) down-regulates hepatitis B virus X promoter activity by the competition for the activating protein 1 binding site and the formation of the ATF2-Jun heterodimer. AB - The hepatitis B viral X promoter is known to be positively autoregulated by its own HBx protein, which also interacts with many cellular regulatory proteins. We investigated the effect of activating transcription factor 2 (ATF2) on the activity of the X promoter. Cotransfection of the ATF2 expression vector with a X promoter-chloramphenicol acetyltransferase plasmid repressed the X promoter activity in HepG2 cells. HBx activated activating protein 1 (AP-1)-mediated transcription through the hepatitis B virus E element by 35-fold, while its activation activity was inhibited in the presence of ATF2, suggesting that ATF2 inhibited the autoactivation of X promoter by HBx and basal transcription mediated by AP-1. Since the binding sites of AP-1 and ATF2 in the hepatitis B virus E element overlap, the repression of X promoter activity by ATF2 is exerted by the competition for the AP-1 binding site and the formation of the ATF2-Jun heterodimer as in the case of the consensus AP-1 element. However, the small X promoter had a ATF2 binding site and was activated by ATF2. These results suggest that the syntheses of X proteins are differentially regulated by ATF2. PMID- 9202005 TI - Inhibition of Plasmodium falciparum proliferation in vitro by ribozymes. AB - Catalytic RNA (ribozymes) suppressed the growth of the human malarial parasite Plasmodium falciparum in vitro. The phosphorothioated hammerhead ribozymes targeted unique regions of the P. falciparum carbamoyl-phosphate synthetase II gene. The P. falciparum carbamoyl-phosphate synthetase II gene encodes the first and limiting enzyme in the pathway, and its mRNA transcript contains two large insert regions absent in other carbamoyl-phosphate synthetases, including that from humans. These inserts are ideal targets for nucleic acid therapy. Exogenous delivery of ribozymes to cultures reduced malarial viability up to 55% at 0.5 microM ribozyme concentrations, which is significantly greater than control levels (5-15% reduction), suggesting a sequence-specific inhibition. This inhibition was shown to be stage-specific, with optimal inhibitions being detected after 24 h, coincident with maximal production of the carbamoyl phosphate synthetase enzyme in the course of the life cycle of the parasite. A decrease in total carbamoyl-phosphate synthetase activity was observed only in cultures treated with the ribozymes. The task of developing alternative therapeutic agents against malaria is urgent due to the evolution of drug resistant strains of P. falciparum, the most virulent of all human malarial parasites. Another critical issue to be addressed is the possibility of eliminating or reducing any systemic toxicity to the host, which can potentially be provided by nucleic acid therapy. This work is the first reported assessment of the ability of ribozymes as antimalarials. Ribozyme inhibition assays can also aid in identifying important antimalarial loci for chemotherapy. The malarial parasite can, in turn, be a useful in vivo host to study the catalysis and function of new ribozyme designs. PMID- 9202006 TI - Role of the glycine triad in the ATP-binding site of cAMP-dependent protein kinase. AB - A glycine-rich loop in the ATP-binding site is one of the most highly conserved sequence motifs in protein kinases. Each conserved glycine (Gly-50, Gly-52, and Gly-55) in the catalytic (C) subunit of cAMP-dependent protein kinase (cAPK) was replaced with Ser and/or Ala. Active mutant proteins were expressed in Escherichia coli, purified to apparent homogeneity, separated into phosphoisoforms, and characterized. Replacing Gly-55 had minimal effects on steady-state kinetic parameters, whereas replacement of either Gly-50 or Gly-52 had major effects on both Km and kcat values consistent with the prediction of the importance of the tip of the glycine-rich loop for catalysis. Substitution of Gly-50 caused a 5-8-fold reduction in Km (ATP), an 8-12-fold increase in Km (peptide), and a 3-5-fold drop in kcat. The Km (ATP) and Km (peptide) values of C(G52S) were increased 8- and 18-fold, respectively, and the kcat was decreased 6 fold. In contrast to catalytic efficiency, the ATPase rates of C(G50S) and C(G52S) were increased by more than an order of magnitude. The thermostability of each mutant was slightly increased. Unphosphorylated C(G52S) was characterized as well as several isoforms phosphorylated at a single site, Ser-338. All of these phosphorylation-defective mutants displayed a substantial decrease in both enzymatic activity and thermal stability that correlated with the missing phosphate at Thr-197. These results are correlated with the crystal structure, models of the respective mutant proteins, and conservation of the Glys within the protein kinase family. PMID- 9202007 TI - Structural and functional complementation of an inactive Bcl-2 mutant by Bax truncation. AB - Interactions among proteins in the Bcl-2 family regulate the onset of programmed cell death. Previous work has shown that the death-inhibiting family members Bcl 2 and Bcl-xL form heterodimers with the death-promoting homologue Bax and that certain site-directed mutants of Bcl-2 and Bcl-xL lose both biological activity and the ability to bind Bax. To better understand the structural basis of heterodimer formation, we have used a yeast two-hybrid assay to screen for mutants of Bax that regain the ability to bind to these inactive Bcl-2(G145A) and Bcl-xL(G138A) mutants. This screen identified a series of C-terminally truncated Bax molecules that contain complete BH3 (Bcl-2 homology domain 3) domains but that have lost BH1 and BH2 sequences. These results indicate that while the Bcl-2 and Bcl-xL mutants fail to bind full-length Bax, they still retain a binding site for the critical BH3 domain. This suggests that conformational constraints in full-length Bax regulate its ability to bind to other Bcl-2 family members. Furthermore, we demonstrate that the normally inert Bcl-2(G145A) mutant effectively blocks apoptosis induced by a C-terminally truncated Bax molecule, but does not block apoptosis induced by wild-type Bax. This demonstrates that cell protection can be effected by directly binding pro-apoptotic members of the Bcl-2 family. PMID- 9202008 TI - Allosteric mechanism of induction of CytR-regulated gene expression. Cytr repressor-cytidine interaction. AB - Transcription from cistrons of the Escherichia coli CytR regulon is activated by E. coli cAMP receptor protein (CRP) and repressed by a multiprotein complex composed of CRP and CytR. De-repression results when CytR binds cytidine. CytR is a homodimer and a LacI family member. A central question for all LacI family proteins concerns the allosteric mechanism that couples ligand binding to the protein-DNA and protein-protein interactions that regulate transcription. To explore this mechanism for CytR, we analyzed nucleoside binding in vitro and its coupling to cooperative CytR binding to operator DNA. Analysis of the thermodynamic linkage between sequential cytidine binding to dimeric CytR and cooperative binding of CytR to deoP2 indicates that de-repression results from just one of the two cytidine binding steps. To test this conclusion in vivo, CytR mutants that have wild-type repressor function but are cytidine induction deficient (CID) were identified. Each has a substitution for Asp281 or neighboring residue. CID CytR281N was found to bind cytidine with three orders of magnitude lower affinity than wild-type CytR. Other CytR mutants that do not exhibit the CID phenotype were found to bind cytidine with affinity similar to wild-type CytR. The rate of transcription regulated by heterodimeric CytR composed of one CytR281N and one wild-type subunit was compared with that regulated by wild-type CytR under inducing conditions. The data support the conclusion that the first cytidine binding step alone is sufficient to induce. PMID- 9202009 TI - The yeast Rab escort protein binds intracellular membranes in vivo and in vitro. AB - In both mammals and yeast, intracellular vesicular transport depends on the correct shuttling between membrane and cytosol of the Rab/Ypt small G proteins. Membrane association of these proteins requires prenylation by the Rab geranylgeranyl transferase that recognizes a complex formed by the Rab/Ypt protein and the Rab escort protein (REP). After prenylation the Rab/Ypt protein is delivered to the target membranes by REP. Little is known about the early steps of the Rab-REP complex formation and where this association occurs in the cell. Although prenylation is believed to take place in the cytosol, we show that the yeast Rab escort protein Mrs6 is present in both soluble and particulate fractions of cell extracts. Mrs6p is associated with the heavy microsomal fraction that contains endoplasmic reticulum-Golgi membranes but is absent in the plasma membrane, vacuoles, mitochondria, and microsomal subfraction associated with mitochondria. The solubilization pattern of the particulate pool of Mrs6p implies that this protein is peripherally but tightly associated with membranes via hydrophobic interactions and metal ions. We also report that the C terminus of Mrs6p is important for maintaining the solubility of the protein because its deletion or replacement with the C terminus of RabGDI results in a protein that localizes only to membranes. PMID- 9202010 TI - Depolarization of rat brain synaptosomes increases phosphorylation of voltage sensitive sodium channels. AB - Depolarization of rat brain synaptosomes causes an increase in phosphorylation of serine residues 573, 610, 623, and 687 on voltage-sensitive sodium channels. Although these sites have been shown to be phosphorylated by cAMP-dependent protein kinase in vitro and in situ, the depolarization-induced increase in their state of phosphorylation is not due to increased cAMP-dependent protein kinase activity, but requires calcium influx and protein kinase C. Since phosphorylation at this cluster of sites inhibits sodium current and would decrease neuronal excitability, this may be an important negative feedback mechanism whereby calcium influx during prolonged or repetitive depolarization can attenuate neuronal excitability and prevent further calcium accumulation. Phosphorylation of purified channels by protein kinase C decreases dephosphorylation of cAMP dependent phosphorylation sites by purified calcineurin or protein phosphatase 2A. This suggests that one mechanism by which protein kinase C may increase phosphorylation of cAMP-dependent phosphorylation sites in sodium channels is to inhibit their dephosphorylation. This represents an important new mechanism for convergent regulation of an ion channel by two distinct signal transduction pathways. PMID- 9202011 TI - Cyclic ADP-ribose-gated Ca2+ release in sea urchin eggs requires an elevated. AB - Cyclic ADP-ribose (cADPr) has been shown to release intracellular Ca2+ from sea urchin eggs and a variety of vertebrate cell types, although its mechanism of action remains elusive. We employed the caged version of cADPr to study the [Ca2+] transient kinetics in intact sea urchin eggs for insights into how cADPr gates Ca2+ release. Ca2+ release triggered by photolytic production of cADPr was initially slow, with an effective delay of several hundred milliseconds before the onset of a rapid Ca2+ release phase. In contrast, Ca2+ release induced by photolysis of caged inositol 1,4,5-trisphosphate was immediate in onset and roughly an order of magnitude faster. The delay before cADPr-induced Ca2+ release was eliminated when the [Ca2+] was step-elevated coincident with the photoliberation of cADPr and greatly prolonged in the presence of exogenous Ca2+ buffers. Thus, the slow onset of Ca2+ release does not reflect an intrinsically slow rate by which cADPr gates release channels. Rather, a [Ca2+] rise from resting levels is needed to achieve more than minimal cADPr activity. Full release of Ca2+ by cADPr in intact sea urchin eggs requires a positive Ca2+ feedback. PMID- 9202013 TI - Macrophage stimulating protein (MSP) binds to its receptor via the MSP beta chain. AB - Macrophage stimulating protein (MSP) is a 78-kDa disulfide-linked heterodimer belonging to the plasminogen-related kringle protein family. MSP activates the RON receptor protein-tyrosine kinase, which results in cell migration, shape change, or proliferation. A structure-activity study of MSP was performed using pro-MSP, MSP, MSP alpha and beta chains, and a complex including the first two kringles and IgG Fc (MSP-NK2). Radioiodinated MSP and MSP beta chain both bound specifically to RON. The Kd of 1.4 nM for MSP beta chain is higher than the reported Kd range of 0.6-0.8 nM for MSP. Pro-MSP, MSP alpha chain, and MSP-NK2 did not bind. Only MSP stimulated RON autophosphorylation. Although the beta chain bound to RON and partially inhibited MSP-induced RON phosphorylation in kidney 293 cells, it did not induce RON phosphorylation. Pro-MSP, MSP alpha chain, or MSP-NK2 failed to activate RON, consistent with their inability to bind to the RON receptor. Functional studies showed that only MSP induced cell migration, and shape change in resident macrophages, and growth of murine keratinocytes. Our data indicate that the primary receptor binding domain is located in a region of the MSP beta chain, in contrast to structurally similar hepatocyte growth factor, in which the receptor binding site is in the alpha chain. However, full activation of RON requires binding of the complete MSP disulfide-linked alphabeta chain heterodimer. PMID- 9202012 TI - p-Hydroxyphenylacetaldehyde, the major product of L-tyrosine oxidation by the myeloperoxidase-H2O2-chloride system of phagocytes, covalently modifies epsilon amino groups of protein lysine residues. AB - Activated human phagocytes employ the myeloperoxidase-H2O2-Cl- system to convert L-tyrosine to p-hydroxyphenylacetaldehyde (pHA). We have explored the possibility that pHA covalently reacts with proteins to form Schiff base adducts, which may play a role in modifying targets at sites of inflammation. Because Schiff bases are labile to acid hydrolysis, prior to analysis the adducts were rendered stable by reduction with NaCNBH3. Purified pHA reacted with Nalpha-acetyllysine, an analog of protein lysine residues. The reduced reaction product was identified as Nalpha-acetyl-Nepsilon-(2-(p-hydroxyphenyl)ethyl)lysine by 1H NMR spectroscopy and mass spectrometry. The compound Nepsilon-(2-(p-hydroxyphenyl)ethyl)lysine (pHA-lysine) was likewise identified in acid hydrolysates of bovine serum albumin (BSA) that were first exposed to myeloperoxidase, H2O2, L-tyrosine, and Cl- and then reduced with NaCNBH3. Other halides (F-, Br-, I-) and the pseudohalide SCN- could not replace Cl- as a substrate in the myeloperoxidase-H2O2-L-tyrosine system. In the absence of the enzymatic system, pHA-lysine was detected in reduced reaction mixtures of BSA, L-tyrosine, and reagent HOCl. In contrast, pHA lysine was undetectable when BSA was incubated with L-tyrosine and HOBr, peroxynitrite, hydroxyl radical, or a variety of other peroxidases, indicating that the aldehyde-protein adduct was selectively produced by HOCl. Human neutrophils activated in the presence of tyrosine also modified BSA lysine residues. pHA-lysine formation required L-tyrosine and cell activation; it was inhibited by peroxidase inhibitors and catalase, implicating myeloperoxidase and H2O2 in the reaction pathway. pHA-lysine was detected in inflamed human tissues that were reduced, hydrolyzed, and then analyzed by mass spectrometry, indicating that the reaction of pHA with proteins may be of physiological importance. These observations raise the possibility that the identification of pHA-lysine in tissues will pinpoint targets where phagocytes inflict oxidative damage in vivo. PMID- 9202014 TI - Abeta(1-40) prevents heparanase-catalyzed degradation of heparan sulfate glycosaminoglycans and proteoglycans in vitro. A role for heparan sulfate proteoglycan turnover in Alzheimer's disease. AB - Alzheimer's disease is characterized by senile plaques composed of polymeric fibrils of beta amyloid (Abeta), a 39-42-amino acid peptide formed after proteolytic processing of the amyloid precursor protein (betaAPP). Heparan sulfate proteoglycans have been shown to colocalize with Abeta in Alzheimer's disease brain, and experimental evidence indicates that the interactions between the proteoglycan and the peptide are important for the promotion, deposition, and/or persistence of the senile plaques. Studies in rat brain indicated that both the core protein and the heparan sulfate glycosaminoglycan chains are required for amyloid fiber formation and deposition in vivo (Snow, A. D., Sekiguchi, R., Nochlin, D., Fraser, P., Kimata, K. , Mizutani, A., Arai, M., Schreier, W. A., and Morgan, D. G. (1994) Neuron 12, 219-234), suggesting that one mechanism to prevent the formation of Abeta-heparan sulfate proteoglycan complexes that lead to deposition of amyloid would be to degrade the proteoglycan. Normally, heparan sulfate proteoglycans are internalized and degraded to short glycosaminoglycans by intracellular heparanases. These reactions occur in the endosomal-lysosomal pathway, which is the same intracellular location where betaAPP is processed to Abeta. Using partially purified heparanase activities from Chinese hamster ovary cells we examined whether Abeta(1-40) affects the catabolism of Chinese hamster ovary heparan sulfate glycosaminoglycans and proteoglycans in vitro. Abeta(1-40) binds to both the long heparan sulfate glycosaminoglycans attached to core proteins and the short, heparanase-derived chains in a concentration-dependent and pH-dependent manner. When Abeta(1-40) is added to heparanase assays, it prevents the partially purified activities from releasing heparan sulfate chains from core proteins and degrading them to short glycosaminoglycans; however, a large molar excess of the peptide to heparan sulfate is required to see the effect. Our results suggest that normally the levels of Abeta in the endosomal pathway are not sufficient to interfere with heparanase activity in vivo. However, once the level of Abeta peptides are elevated, as they are in Alzheimer's disease, they could interact with heparan sulfate proteoglycans and prevent their catabolism. This could promote the formation and deposition of amyloid, since the binding of Abeta to the proteoglycan species will predominate. PMID- 9202015 TI - Homocyst(e)ine decreases bioavailable nitric oxide by a mechanism involving glutathione peroxidase. AB - Hyperhomocyst(e)inemia is believed to injure endothelial cells in vivo through a number of mechanisms, including the generation of hydrogen peroxide (H2O2). Earlier in vitro studies demonstrated that homocyst(e)ine (Hcy) decreases the biological activity of endothelium-derived relaxing factor and that this decrease can be reversed by preventing the generation of hydrogen peroxide. Here we show that Hcy treatment of bovine aortic endothelial cells leads to a dose-dependent decrease in NOx (p = 0.001 by one-way analysis of variance) independent of endothelial nitric-oxide synthase activity or protein levels and nos3 transcription, suggesting that Hcy affects the bioavailability of NO, not its production. We hypothesized that, in addition to increasing the generation of H2O2, Hcy decreases the cell's ability to detoxify H2O2 by impairing intracellular antioxidant enzymes, specifically the intracellular isoform of glutathione peroxidase (GPx). To test this hypothesis, confluent bovine aortic endothelial cells were treated with a range of concentrations of Hcy, and intracellular GPx activity was determined. Compared with control cells, cells treated with Hcy showed a significant reduction in GPx activity (up to 81% at 250 microM Hcy). In parallel with the decrease in GPx activity, steady-state GPx mRNA levels were also significantly decreased compared with control levels after exposure to Hcy, which appeared not to be a consequence of message destabilization. These data suggest a novel mechanism by which Hcy, in addition to increasing the generation of hydrogen peroxide, may selectively impair the endothelial cell's ability to detoxify H2O2, thus rendering NO more susceptible to oxidative inactivation. PMID- 9202016 TI - The fate of cholesterol exiting lysosomes. AB - Cholesterol released from ingested low density lipoproteins in lysosomes moves both to the plasma membrane and to the endoplasmic reticulum (ER) where it is re esterified. Whether cholesterol can move directly from lysosomes to ER or first must traverse the plasma membrane has not been established. To examine this question, the endocytic pathway of rat hepatoma cells was loaded at 18 degrees C with low density lipoproteins (LDL) labeled with [3H]cholesteryl linoleate, and the label then was chased at 37 degrees C. The hydrolysis of the accumulated ester proceeded linearly for several hours. Almost all of the released [3H]cholesterol moved to the plasma membrane rapidly and without a discernable lag. In contrast, the re-esterification in the ER of the released [3H]cholesterol showed a characteristic lag of 0.5-1 h. These data are inconsistent with direct cholesterol transfer from lysosomes to ER; rather, they suggest movement through the plasma membrane. Furthermore, we found that progesterone, imipramine and 3 beta-[2-(diethylamino)ethoxy]androst-5-en-17-one (U18666A) strongly inhibited the re-esterification of lysosomal cholesterol in the ER. However, contrary to previous reports, they did not block transfer of [3H]cholesterol from lysosomes to the cell surface. Therefore, the site of action of these agents was not at the lysosomes. We suggest instead that their known ability to block cholesterol movement from the plasma membrane to the ER accounts for the inhibition of lysosomal cholesterol esterification. These findings are consistent with the hypothesis that cholesterol released from lysosomes passes through the plasma membrane on its way to the ER rather than proceeding there directly. As a result, ingested cholesterol is subject to the same homeostatic regulation as the bulk of cell cholesterol, which is located in the plasma membrane. PMID- 9202017 TI - Seminiferous tubule basement membrane. Composition and organization of type IV collagen chains, and the linkage of alpha3(IV) and alpha5(IV) chains. AB - Seminiferous tubule basement membrane (STBM) plays an important role in spermatogenesis. In the present study, the composition and structural organization of type IV collagen of bovine STBM was investigated. STBM was found to be composed of all six alpha-chains of type IV collagen based upon immunocytochemical and biochemical analysis. The content of alpha3(IV) chain (40%) and the alpha4(IV) chain (18%) was substantially higher than in any other basement membrane collagen. The supramolecular structure of the six alpha(IV) chains was investigated using pseudolysin (EC 3.4.24.26) digestion to excise triple-helical molecules, subsequent collagenase digestion to produce NC1 hexamers and antibody affinity chromatography to resolve populations of NC1 hexamers. The hexamers, which reflect specific arrangements of alpha(IV) chains, were characterized for their alpha(IV) chain composition using high performance liquid chromatography, two-dimensional electrophoresis, and immunoblotting with alpha(IV) chain-specific antibodies. Three major hexamer populations were found that represent the classical network of the alpha1(IV) and alpha2(IV) chains and two novel networks, one composed of the alpha1(IV)-alpha6(IV) chains and the other composed of the alpha3(IV)-alpha6(IV) chains. The results establish a structural linkage between the alpha3(IV) and alpha5(IV) chains, suggesting a molecular basis for the conundrum in which mutations in the gene encoding the alpha5(IV) chain cause defective assembly of the alpha3(IV) chain in the glomerular basement membrane of patients with Alport syndrome. PMID- 9202018 TI - Specificity for activase is changed by a Pro-89 to Arg substitution in the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase. AB - Tobacco activase does not markedly facilitate the activation of ribulose-1,5 bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1. 39) from non-Solanaceae species, including the green alga Chlamydomonas reinhardtii. To examine the basis of this specificity, we focused on two exposed residues in the large subunit of Rubisco that are unique to the Solanaceae proteins. By employing in vitro mutagenesis and chloroplast transformation, P89R and K356Q substitutions were separately made in the Chlamydomonas enzyme to change these residues to those present in tobacco. Both mutants were indistinguishable from the wild type when grown with minimal medium in the light and contained wild-type levels of holoenzyme. Purified Rubisco was assessed for facilitated activation by spinach and tobacco activase. Both wild-type and K356Q Rubisco were similar in that spinach activase was much more effective than tobacco activase. In contrast, P89R Rubisco was not activated by spinach activase but was well activated by tobacco activase. Thus, the relative specificities of the spinach and tobacco activases for Chlamydomonas Rubisco were switched by changing a single residue at position 89. This result provides evidence for a site on the Rubisco holoenzyme that interacts directly with Rubisco activase. PMID- 9202019 TI - Regulation of the AKAP79-protein kinase C interaction by Ca2+/Calmodulin. AB - The A kinase-anchoring protein AKAP79 coordinates the location of the cAMP dependent protein kinase (protein kinase A), calcineurin, and protein kinase C (PKC) at the postsynaptic densities in neurons. Individual enzymes in the AKAP79 signaling complex are regulated by distinct second messenger signals; however, both PKC and calcineurin are inhibited when associated with the anchoring protein, suggesting that additional regulatory signals must be required to release active enzyme. This report focuses on the regulation of AKAP79-PKC interaction by calmodulin. AKAP79 binds calmodulin with high affinity (KD of 28 +/- 4 nM (n = 3)) in a Ca2+-dependent manner. Immunofluorescence staining shows that both proteins exhibit overlapping staining patterns in cultured hippocampal neurons. Calmodulin reversed the inhibition of PKCbetaII by the AKAP79(31-52) peptide and reduced inhibition by the full-length AKAP79 protein. The effect of calmodulin on inhibition of a constitutively active PKC fragment by the AKAP79(31 52) peptide was shown to be partially dependent on Ca2+. Ca2+/calmodulin reduced PKC coimmunoprecipitated with AKAP79 and resulted in a 2.6 +/- 0.5-fold (n = 6) increase in PKC activity in a preparation of postsynaptic densities. Collectively, these findings suggest that Ca2+/calmodulin competes with PKC for binding to AKAP79, releasing the inhibited kinase from its association with the anchoring protein. PMID- 9202020 TI - Thioredoxin reductase-dependent inhibition of MCB cell cycle box activity in Saccharomyces cerevisiae. AB - Mlu1 cell cycle box (MCB) elements are found near the start site of yeast genes expressed at G1/S. Basal promoters dependent on the elements for upstream activating sequence activity are inactive in Deltaswi6 yeast. Yeast were screened for mutations that activated MCB reporter genes in the absence of Swi6. The mutations identified a single complementation group. Functional cloning revealed the mutations were alleles of the TRR1 gene encoding thioredoxin reductase. Although deletion of TRR1 activated MCB reporter genes, high copy expression did not suppress reporter gene activity. The trr1 mutations strongly (20-fold) stimulated MCB- and SCB (Swi4/Swi6 cell cycle box)-containing reporter genes, but also weakly (3-fold) stimulated reporter genes that lacked these elements. The trr1 mutations did not affect the level or periodicity of three endogenous MCB gene mRNAs (TMP1, RNR1, and SWI4). Deletion of thioredoxin genes TRX1 and TRX2 recapitulated the stimulatory effect of trr1 mutations on MCB reporter gene activity. Conditions expected to oxidize thioredoxin (exposure to H2O2) induced MCB gene expression, whereas conditions expected to conserve thioredoxin (exposure to hydroxyurea) inhibited MCB gene expression. The results suggest that thioredoxin oxidation contributes to MCB element activation and suggest a link between thioredoxin-oxidizing processes such as ribonucleotide reduction and cell cycle-specific gene transcription. PMID- 9202021 TI - Decreased endosomal delivery of major histocompatibility complex class II invariant chain complexes in dynamin-deficient cells. AB - Major histocompatibility complex class II molecules are heterodimeric cell surface molecules which acquire antigenic peptides in the endosomal/lysosomal system. Invariant chain (Ii), a third chain which is associated with class II molecules intracellularly mediates the endosomal targeting, but it is debated whether class II molecules reach the endosomal system mainly from the trans-Golgi network or via the cell surface. Dynamin is a cytosolic GTPase which is necessary for the formation of clathrin-coated vesicles from the plasma membrane, but which is not required for vesicle formation from the trans-Golgi network. Here we have used HeLa cells expressing a dominant negative form of dynamin to show that inhibition of clathrin-mediated uptake from the plasma membrane leads to accumulation of transfected Ii-class II complexes at the cell surface, while delivery of such complexes to endosomes/lysosomes is decreased. Our data therefore suggest that in this experimental system the majority of Ii-class II complexes traverse the cell surface before they reach the endosomal system. PMID- 9202022 TI - Structural organization of the major subunits in cyanobacterial photosystem 1. Localization of subunits PsaC, -D, -E, -F, and -J. AB - Based on an improved isolation procedure using perfusion chromatography, trimeric Photosystem 1 (PS1) complexes have been isolated from various deletion mutants of the mesophilic cyanobacterium Synechocystis PCC 6803. These mutants are only deficient in the deleted subunits, which was carefully checked by high resolution gel electrophoresis in combination with immunoblotting. These highly purified and well characterized PS1 particles were then examined by electron microscopy, followed by computer-aided image processing with single particle averaging techniques as described earlier (Kruip, J., Boekema, E. J., Bald, D., Boonstra, A. F., and Rogner, M. (1993) J. Biol. Chem. 268, 23353-23360). This precise methodological approach allowed a confident localization of the PS1 subunits PsaC, -D, -E, -F, and -J; it also shows shape and size of these subunits once integrated in the PS1 complex. Subunits PsaC, -D, and -E form a ridge on the stromal site, with PsaE toward the edge of each monomer within the trimer and PsaD extending toward the trimeric center, leaving PsaC in between. PsaF (near PsaE) and PsaJ are close together on the outer edge of each monomer; their proximity is also supported by chemical cross-linking, using the zero-length cross-linker EDC. This localization of PsaF contradicts the position suggested by the published low resolution x-ray analysis and shows for the first time the existence of at least one transmembrane alpha-helix for PsaF. A topographic three dimensional map has been drawn from this set of results showing the location of the major PS1 subunits (besides PsaA and PsaB). These data also led to the assignment of electron density in the recent medium resolution x-ray structure for PS1 (Krauss, N., Schubert, W.-D., Klukas, O., Fromme, P., Witt, H. T., Saenger, W. (1996) Nat. Struct. Biol. 3, 965-973). PMID- 9202023 TI - Characterization of the WW domain of human yes-associated protein and its polyproline-containing ligands. AB - We had previously identified the WW domain as a novel globular domain that is composed of 38-40 semiconserved amino acids and is involved in mediating protein protein interaction. The WW domain is shared by proteins of diverse functions including structural, regulatory, and signaling proteins in yeast, nematode, and mammals. Functionally it is similar to the Src homology 3 domain in that it binds polyproline ligands. By screening a 16-day mouse embryo expression library, we identified two putative ligands of the WW domain of Yes kinase-associated protein which we named WW domain-binding proteins 1 and 2. These proteins interacted with the WW domain via a short proline-rich motif with the consensus sequence of four consecutive prolines followed by a tyrosine. Herein, we report the cDNA cloning and characterization of the human orthologs of WW domain-binding proteins 1 and 2. The products encoded by these cDNA clones represent novel proteins with no known function. Furthermore, these proteins show no homology to each other except for a proline-rich motif. By fluorescence in situ hybridization on human metaphase chromosomes, we mapped the human genes for WW domain-binding proteins 1 and 2 to chromosomes 2p12 and 17q25, respectively. In addition, using site directed mutagenesis, we determined which residues in the WW domain of Yes kinase associated protein are critical for binding. Finally, by synthesizing peptides in which the various positions of the four consecutive proline-tyrosine motif and the five surrounding residues were replaced by all possible amino acid residues, we further elucidated the binding requirements of this motif. PMID- 9202024 TI - Interaction of the NH2-terminal domain of fibronectin with heparin. Role of the omega-loops of the type I modules. AB - Determinants of the interaction of the 29-kDa NH2-terminal domain of fibronectin with heparin were explored by analysis of normal and mutant recombinant NH2 terminal fibronectin fragments produced in an insect cell Baculovirus host vector system. A genomic/cDNA clone was constructed that specified a secretable human fibronectin NH2 fragment. With the use of site-directed mutagenesis a set of 29 kDa fragments was obtained that contained glycine or glutamic acid residues in place of basic residues at various candidate sites for heparin binding in the five type I modules that make up the domain. The recombinant fragment containing the wild type sequence had a nearly normal circular dichroic spectra and a melting profile, as assayed by loss of ellipticity at 228 nm, that was indistinguishable from that of the native fragment obtained by trypsinization of plasma fibronectin. A substantial proportion of the wild type recombinant fragment bound to heparin-Sepharose, where it was eluted at the same NaCl concentration as the native fragment. The wild type fragment was capable of promoting matrix-driven translocation, a morphogenetic effect in artificial extracellular matrices that depends on the interaction of the fibronectin NH2 terminus with heparin-like molecules on the surfaces of particles. Mutant fragments in which arginines predicted to be most exposed in the folded fragment were converted to glycines retained the same affinity for heparin as the wild type fragment. In contrast, a mutant fragment in which the single basic residue (Arg99) in the minor loop ("Omega-loop") of the second type I module was converted to a glycine had an essentially normal melting profile but exhibited no binding to heparin and failed to promote matrix-driven translocation. A mutant fragment in which the single basic residue (Arg52) of the first type I module was converted to a glycine also completely lacked heparin binding activity, but one in which the single basic residue (Arg191) the fourth type I module was converted to a glycine retained the ability to bind heparin. A mutant fragment in which the single basic residue (Lys143) in the Omega-loop of the third type I module was converted to a glutamic acid lacked heparin binding activity but had a CD spectrum similar to the heparin-liganded native protein and was capable of promoting matrix-driven translocation. The results indicate that multiple residues in the Omega-loops of the fibronectin NH2-terminal domain participate in its interactions with heparin. In addition, the conformation of one of the nonbinding mutants may mimic the heparin-induced structural alteration in this fibronectin domain required for certain morphogenetic events. PMID- 9202025 TI - Nitric oxide trapping of the tyrosyl radical of prostaglandin H synthase-2 leads to tyrosine iminoxyl radical and nitrotyrosine formation. AB - The determination of protein nitrotyrosine content has become a frequently used technique for the detection of oxidative tissue damage. Protein nitration has been suggested to be a final product of the production of highly reactive nitrogen oxide intermediates (e. g. peroxynitrite) formed in reactions between nitric oxide (NO.) and oxygen-derived species such as superoxide. The enzyme prostaglandin H synthase-2 (PHS-2) forms one or more tyrosyl radicals during its enzymatic catalysis of prostaglandin formation. In the presence of the NO. generator diethylamine nonoate, the electron spin resonance spectrum of the PHS-2 derived tyrosyl radical is replaced by the spectrum of another free radical containing a nitrogen atom. The magnitude of the nitrogen hyperfine coupling constant in the latter species unambiguously identifies it as an iminoxyl radical, which is likely formed by the oxidation of nitrosotyrosine, a stable product of the addition of NO. to tyrosyl radical. Addition of superoxide dismutase did not alter the spectra, indicating that peroxynitrite was not involved. Western blot analysis of PHS-2 after exposure to the NO.-generator revealed nitrotyrosine formation. The results provide a mechanism for nitric oxide-dependent tyrosine nitration that does not require formation of more highly reactive nitrogen oxide intermediates such as peroxynitrite or nitrogen dioxide. PMID- 9202026 TI - Recombination-dependent repair of DNA double-strand breaks with purified proteins from Escherichia coli. AB - We have developed an in vitro system in which repair of DNA double-strand breaks is performed by purified proteins of Escherichia coli. A segment was deleted from a circular duplex DNA molecule by restriction at two sites. 3' single-stranded overhangs were introduced at both ends of the remaining linear fragment. In a first step, RecA protein catalyzed the formation of a D-loop between one single stranded tail and a homologous undeleted supercoiled DNA molecule. In a second step, E. coli DNA polymerase II or III used the 3' end in the D-loop as a primer to copy the missing sequences of the linear substrate on one strand of the supercoiled template. Under proper conditions, the integrity of the deleted substrate was restored, as shown by analysis of the products by electrophoresis, restriction, and transformation. In this reaction, DNA synthesis is strictly dependent on recombination, and repair is achieved without formation of a Holliday junction. PMID- 9202028 TI - Ubiquitous expression of the alpha1(XIX) collagen gene (Col19a1) during mouse embryogenesis becomes restricted to a few tissues in the adult organism. AB - Type XIX collagen is a poorly characterized member of the fibril-associated collagens with an interrupted triple helices (FACIT) class of collagen molecules. As a first step toward elucidating its function, we have isolated full size cDNA clones from the mouse alpha1(XIX) collagen gene (Col19a1) and established its pattern of expression in the developing embryo and adult organism. Col19a1 transcripts can be detected as early as 11 days of gestation and in all embryonic tissues, except the liver, of an 18-day postcoitum mouse. In contrast, only a few adult tissues, brain, eye, and testis, seem to accumulate Col19a1 mRNA. Col19a1 transcripts are at least 10 times more abundant in adult than fetal brain and significantly less in adult than fetal muscle and skin. Consistent with the RNA data, polyclonal antibodies for alpha1(XIX) collagen reacted with a 150-kDa protein in the neutral salt extraction of adult mouse brain tissues. We therefore propose that type XIX collagen plays a distinct role from the other FACIT molecules, particularly in the assembly of embryonic matrices and in the maintenance of specific adult tissues. PMID- 9202027 TI - Vasoactive peptides modulate vascular endothelial cell growth factor production and endothelial cell proliferation and invasion. AB - The proliferation of vascular endothelial cells (EC) is an important event in angiogenesis. The synthesis of the EC growth factor, vascular endothelial cell growth factor (VEGF), is stimulated by a variety of activators; but the effects of important vasoactive peptides are not well understood, and there are no known natural inhibitors of VEGF production. We found that the vasoactive peptides endothelin (ET)-1 and ET-3 stimulated the synthesis of VEGF protein 3-4-fold in cultured human vascular smooth muscle cells, comparable in magnitude to hypoxia. ET-1 and ET-3 acted through the ETA and ETB receptors, respectively, and signaling through protein kinase C was important. Atrial natriuretic peptide (ANP), C-type natriuretic peptide, and C-ANP-(4-23), a ligand for the natriuretic peptide clearance receptor, equipotently inhibited production of VEGF by as much as 88% and inhibited ET- or hypoxia-stimulated VEGF transcription. EC proliferation and invasion of matrix were stimulated by VEGF secreted into the medium by ET-incubated vascular smooth muscle cells. This was inhibited by ANP. Our results identify the natriuretic peptides as the first peptide inhibitors of VEGF synthesis and indicate a novel mechanism by which vasoactive peptides could modulate angiogenesis. PMID- 9202029 TI - Structure of the m1 muscarinic acetylcholine receptor gene and its promoter. AB - The m1 receptor is one of five muscarinic receptors that mediate the metabotropic actions of acetylcholine in the nervous system where it is expressed predominantly in the telencephalon and autonomic ganglia. RNase protection, primer extension, and 5'-rapid amplification of cDNA ends analysis of a rat cosmid clone containing the entire m1 gene demonstrated that the rat m1 gene consists of a single 657-base pairs (bp) non-coding exon separated by a 13. 5 kilobase (kb) intron from a 2.54-kb coding exon that contains the entire open reading frame. The splice acceptor for the coding exon starting at -71 bp relative to the adenine of the initiating methionine. This genomic structure is similar to that of the m4 gene (Wood, I. C., Roopra, A., Harrington, C. A., and Buckley, N. J. (1995) J. Biol. Chem. 270, 30933-30940 and Wood, I. C., Roopra, A., and Buckley, N. J. (1996) J. Biol. Chem. 271, 14221-14225). Like the m4 gene, the m1 promoter lacks TATA and CAAT consensus motifs, and the first exon and 5' flanking region are not gc-rich. The 5'-flanking region also contains the consensus regulatory elements Sp-1, NZF-1, AP-1, AP-2, E-box, NFkappaB, and Oct 1. Unike the m4 promoter, there is no evidence of a RE1/NRSE silencer element in the m1 promoter. Deletional analysis and transient transfection assays demonstrates that reporter constructs containing 0.9 kb of 5'-flanking sequence and the first exon are sufficient to drive cell-specific expression of reporter gene in IMR32 neuroblastoma cells while remaining silent in 3T3 fibrobasts. PMID- 9202030 TI - Paclitaxel-resistant human ovarian cancer cells have mutant beta-tubulins that exhibit impaired paclitaxel-driven polymerization. AB - Acquired resistance to paclitaxel can be mediated by P-glycoprotein or by alterations involving tubulin. We report two paclitaxel-resistant sublines derived from 1A9 human ovarian carcinoma cells. Single-step paclitaxel selection with verapamil yielded two clones that are resistant to paclitaxel and collaterally sensitive to vinblastine. The resistant sublines are not paclitaxel dependent, and resistance remained stable after 3 years of drug-free culture. All cell lines accumulate [3H]paclitaxel equally, and no MDR-1 mRNA was detected by polymerase chain reaction following reverse transcription. Total tubulin content is similar, but the polymerized fraction increased in parental but not in resistant cells following the paclitaxel addition. Purified tubulin from parental cells demonstrated paclitaxel-driven increased polymerization, in contrast to resistant cell tubulin, which did not polymerize under identical conditions. In contrast, epothilone B, an agent to which the resistant cells retained sensitivity, increased assembly. Comparable expression of beta-tubulin isotypes was found in parental and resistant cells, with predominant expression of the M40 and beta2 isotypes. Sequence analysis demonstrated acquired mutations in the M40 isotype at nucleotide 810 (T --> G; Phe270 --> Val) in 1A9PTX10 cells and nucleotide 1092 (G --> A; Ala364 --> Thr) in 1A9PTX22 cells. These results identify residues beta270 and beta364 as important modulators of paclitaxel's interaction with tubulin. PMID- 9202031 TI - Structure and folding of nascent polypeptide chains during protein translocation in the endoplasmic reticulum. AB - To investigate the role of protein folding and chaperone-nascent chain interactions in translocation across the endoplasmic reticulum membrane, the translocation of wild type and mutant forms of preprolactin were studied in vivo and in vitro. The preprolactin mutant studied contains an 18-amino acid substitution at the amino terminus of the mature protein, eliminating a disulfide bonded loop domain. In COS-7 cells, mutant prolactin accumulated in the endoplasmic reticulum as stable protein-protein and disulfide-bonded aggregates, whereas wild type prolactin was efficiently secreted. In vitro, wild type and mutant preprolactin translocated with equal efficiency although both translation products were recovered as heterogeneous aggregates. Studies with translocation intermediates indicated that aggregation occurred co-translationally. To evaluate the contribution of lumenal chaperones to translocation and folding, in vitro studies were performed with native and reconstituted, chaperone-deficient membranes. The absence of lumenal chaperones was associated with a decrease in translocation efficiency and pronounced aggregation of the translation products. These studies suggest that chaperone-nascent chain interactions significantly enhance translocation and indicate that in the absence of such interactions, aggregation can serve as the predominant in vitro protein folding end point. The ramifications of these observations on investigations into the mechanism of translocation are discussed. PMID- 9202032 TI - The synaptobrevin-related domains of Bos1p and Sec22p bind to the syntaxin-like region of Sed5p. AB - SNAREs (soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors) are cytoplasmically oriented membrane proteins that reside on vesicular carriers (v-SNARE) and target organelles (t-SNARE). The pairing of a stage-specific v-SNARE with its cognate t-SNARE may mediate the specificity of membrane traffic. In the yeast Saccharomyces cerevisiae transport between the endoplasmic reticulum and Golgi complex employs two v-SNAREs, Bos1p and Sec22p, each containing a domain that is related to the neuronal v-SNARE synaptobrevin. Sed5p, which is homologous to syntaxin, is the t-SNARE that functions at this stage of the secretory pathway. Here we report that regions of Bos1p and Sec22p, which are homologous to synaptobrevin, bind to the syntaxin-like domain of Sed5p. Furthermore, we demonstrate that efficient v-SNARE/t-SNARE interactions require the participation of both v-SNAREs, indicating that, unlike post-Golgi membrane traffic, the active form of the endoplasmic reticulum to Golgi v-SNARE is a heteromeric complex. PMID- 9202033 TI - A Ni2+ binding motif is the basis of high affinity transport of the Alcaligenes eutrophus nickel permease. AB - Amino acid exchanges in the Alcaligenes eutrophus nickel permease (HoxN) were constructed by site-directed mutagenesis, and their effects on nickel ion uptake were investigated. Mutant hoxN alleles were expressed in Escherichia coli, and activity of the altered permeases was examined via a recently described physiological assay (Wolfram, L., Friedrich, B., and Eitinger, T. (1995) J. Bacteriol. 177, 1840-1843). Replacement of Cys-37, Cys-256, or Cys-318 by alanine did not severely affect nickel ion uptake. This activity of a C331A mutant was diminished by 60%, and a similar phenotype was obtained with a cysteine-less mutant harboring four Cys to Ala exchanges. Alterations in a histidine-containing sequence motif (His-62, Asp-67, His-68), which is conserved in microbial nickel transport proteins, strongly affected or completely abolished transport activity in the E. coli system. The analysis of HoxN alkaline phosphatase fusion proteins implied that His-62, Asp-67, and His-68 exchanges did not interfere with overall membrane topology or stability of the nickel permease. These mutations were reintroduced into the A. eutrophus wild-type strain. Analyses of the resulting HoxN mutants indicated that exchanges in the histidine motif led to a clearly decreased affinity of the permease for nickel ion. PMID- 9202034 TI - P11, a unique member of the S100 family of calcium-binding proteins, interacts with and inhibits the activity of the 85-kDa cytosolic phospholipase A2. AB - Using a two hybrid system screen of a human cDNA library, we have found that p11, a unique member of the S100 family of calcium-binding proteins, interacts with the carboxyl region of the 85-kDa cytosolic phospholipase A2 (cPLA2). p11 synthesized in a cell-free system interacts with cPLA2 in vitro. The p11-cPLA2 complex is detectable from a human bronchial epithelial cell line (BEAS 2B). Furthermore, p11 inhibits cPLA2 activity in vitro. Selective inhibition of p11 expression in the BEAS 2B cells by antisense RNA results in an increased PLA2 activity as well as an increased release of prelabeled arachidonic acid. This study demonstrates a novel mechanism for the regulation of cPLA2 by an S100 protein. PMID- 9202035 TI - Serine/threonine kinase activity associated with the cytoplasmic domain of the lymphotoxin-beta receptor in HepG2 cells. AB - The lymphotoxin-beta receptor (LT-betaR) has been shown to be the receptor for the membrane-bound lymphotoxin heterotrimers LTalpha1/beta2 and LTalpha2/beta1. The extracellular domain of LT-betaR shows extensive similarity with members of the tumor necrosis factor receptor family, while its cytoplasmic domain is distinct and lacks any inherent enzymatic activity. This suggests that the interaction of LT-betaR with other molecules might be important for signal transduction. Here we demonstrate the association of a fusion protein, comprising glutathione S-transferase and the cytoplasmic domain of LT-betaR (GST-LT betaR(CD)), with several proteins in the size range 29-80 kDa from HepG2 cell lysates. We present evidence that two of these proteins are serine/threonine kinases, which associate with amino acids 324-377 of the cytoplasmic domain of LT betaR and phosphorylate this receptor. The characteristics of these novel kinases indicate that they are distinct from the previously described tumor necrosis factor receptor-associated kinases. This suggests the presence of novel signal transduction pathway(s) for LT-betaR. PMID- 9202036 TI - Role of the juxtamembrane domains of the transforming growth factor-alpha precursor and the beta-amyloid precursor protein in regulated ectodomain shedding. AB - Although regulated ectodomain shedding is a well known process that affects a large group of transmembrane molecules, it is not clear how the shedding system selects its substrates. Here we investigate the structural requirements for the regulated shedding of two substrates of the general shedding system, the transforming growth factor-alpha precursor, pro-TGF-alpha, and the beta-amyloid precursor protein, beta-APP. The ability of different regions of pro-TGF-alpha or beta-APP to confer susceptibility to the shedding system was tested using as a reporter a transmembrane molecule that is not a substrate of this shedding system. For this purpose we chose the TGF-beta accessory receptor, betaglycan, since genetic and biochemical evidence showed that betaglycan is not a substrate of the shedding system. We determined that replacement of the 14 extracellular amino acids adjacent to the transmembrane region of betaglycan with the corresponding regions of TGF-alpha or beta-APP rendered betaglycan susceptible to ectodomain shedding. These domain swap constructs were cleaved in response to protein kinase C stimulation, and cleavage was prevented by the metalloprotease inhibitor TAPI, both effects being characteristic of the general shedding system. Domain swap constructs containing the transmembrane and/or the cytoplasmic domains of pro-TGF-alpha did not undergo regulated ectodomain cleavage. We conclude that despite a lack of sequence similarity, the extracellular regions of pro-TGF-alpha and beta-APP immediately preceding their transmembrane domains are key determinants of ectodomain shedding. PMID- 9202037 TI - Evidence for a direct interaction between insulin receptor substrate-1 and Shc. AB - Insulin receptor substrate-1 (IRS-1) and Shc are two proteins implicated in intracellular signal transduction. They are activated by an increasing number of extracellular signals, mediated by receptor tyrosine kinases, cytokine receptors, and G protein-coupled receptors. In this study we demonstrate that Shc interacts directly with IRS-1, using the yeast two-hybrid system and an in vitro interaction assay. Deletion analysis of the proteins to map the domains implicated in this interaction shows that the phosphotyrosine binding domain of Shc binds to the region of IRS-1 comprising amino acids 583-661. An in vitro association assay, performed with or without activation of tyrosine kinases, gives evidence that tyrosine phosphorylation of IRS-1 and Shc drastically improves the interaction. Site-directed mutagenesis on IRS-1 583-693 shows that the asparagine, but not the tyrosine residue of the N625GDY628motif domain, is implicated in the IRS-1-Shc-phosphotyrosine binding interaction. Mutation of another tyrosine residue, Tyr608, also induced a 40% decrease in the interaction. This study, describing a phosphotyrosine-dependent interaction between IRS-1 and Shc, suggests that this association might be important in signal transduction. PMID- 9202038 TI - Transcriptional regulation by triiodothyronine of the UDP-glucuronosyltransferase family 1 gene complex in rat liver. Comparison with induction by 3 methylcholanthrene. AB - This study demonstrates that the expression of the phenol UDP glucuronosyltransferase 1 gene (UGT1A1) is regulated at the transcriptional level by thyroid hormone in rat liver. Following 3,5, 3'-triiodo-L-thyronine (T3) stimulation in vivo, there is a gradual increase in the amount of UGT1A1 mRNA with maximum levels reached 24 h after treatment. In comparison, induction with the specific inducer, 3-methylcholanthrene (3-MC), results in maximal levels of UGT1A1 mRNA after 8 h of treatment. In primary hepatocyte cultures, the stimulatory effect of both T3 and 3-MC is also observed. This induction is suppressed by the RNA synthesis inhibitor actinomycin D, indicating that neither inducer acts at the level of mRNA stabilization. Indeed, nuclear run-on assays show a 3-fold increase in UGT1A1 transcription after T3 treatment and a 6-fold increase after 3-MC stimulation. This transcriptional induction by T3 is prevented by cycloheximide in primary hepatocyte cultures, while 3-MC stimulation is only partially affected after prolonged treatment with the protein synthesis inhibitor. Together, these data provide evidence for a transcriptional control of UGT1A1 synthesis and indicate that T3 and 3-MC use different activation mechanisms. Stimulation of the UGT1A1 gene by T3 requires de novo protein synthesis, while 3-MC-dependent activation is the result of a direct action of the compound, most likely via the aromatic hydrocarbon receptor complex. PMID- 9202039 TI - Evidence for the presence of myosin I in the nucleus. AB - We produced and affinity-purified polyclonal antibodies to adrenal myosin I. These antibodies recognize adrenal myosin I by Western blot analysis (116 kDa) and inhibit the actin-activated ATPase activity of purified adrenal myosin I. They also recognize a 120-kDa protein in extracts prepared from many different cell lines. Fluorescence microscopy demonstrated the presence of immunoreactive material in the perinuclear region, the leading edges, and the nuclei of 3T3 cells. Fluorescence microscopy also demonstrated nuclear staining in mouse oocytes at the germinal vesicle stage and in the pronuclei during fertilization. Confocal and immunoelectron microscopy confirmed the intranuclear localization. Electron microscopy also demonstrated staining of structures in nucleoli that are thought to be associated with rDNA transcription. Western blot analyses revealed the presence of the 120-kDa protein in extracts prepared from nuclei that are apparently free of cytosolic contamination. The same nuclear protein binds 125I calmodulin and is photoaffinity labeled with [alpha-32P]ATP. The 120-kDa protein was partially purified from twice washed nuclei using ammonium sulfate fractionation and gel filtration chromatography. Column fractions containing 120 kDa protein as revealed by Western blot analysis also contain K+-EDTA ATPase activity. The 120-kDa protein was also shown to bind actin in the absence, but not the presence, of ATP. Since K+-EDTA ATPase activity, actin, and ATP binding are defining features of the members of the myosin superfamily of proteins, we propose that the 120-kDa protein is a previously undescribed myosin I isoform that is an intranuclear actin-based molecular motor. PMID- 9202040 TI - Induced mutant mice expressing lipoprotein lipase exclusively in muscle have subnormal triglycerides yet reduced high density lipoprotein cholesterol levels in plasma. AB - To determine the contribution of muscle lipoprotein lipase (LPL) to lipoprotein metabolism, induced mutant mice were generated that express human LPL exclusively in muscle. By cross-breeding heterozygous LPL knockout mice with transgenic mice expressing human LPL only in muscle, animals were obtained that express human LPL primarily in skeletal muscle on either the null (L0-MCK) or normal (L2-MCK) LPL backgrounds, and these were compared with control littermates (L2). Fed and fasted post-heparin plasma (PHP) LPL activities were increased 1.4- and 2.3-fold, respectively, in L2-MCK mice and were normal in L0-MCK mice compared with controls. The specific enzyme activities of human LPL in mouse plasma was comparable to human LPL in human PHP. Skeletal muscle LPL activity was increased in both L2-MCK and L0-MCK mice in the fed (6.6-fold) and fasted (4.2-fold in L2 MCK; and 3.4-fold in L0-MCK) states. Adipose tissue LPL mRNA and activity were not detectable in L0-MCK mice. Growth and body mass composition were similar among all groups. In the fasted and fed state, L2-MCK mice had 31% and 53% reductions, respectively, in plasma triglycerides (TG), compatible with increased PHP LPL activity. Unexpectedly, both in the fasted and fed state the L0-MCK mice also had reduced TG (22%), despite normal PHP LPL activities. Very low density lipoprotein (VLDL) turnover studies revealed that the decreased TG were due to increased particle fractional catabolic rate in both L2-MCK and L0-MCK mice. Despite reduced TG, both L2-MCK and L0-MCK mice showed reduced high density lipoprotein (HDL) cholesterol levels (16% and 19%, respectively). HDL turnover studies indicated increased HDL cholesteryl ester fractional catabolic rate in the L2-MCK and L0-MCK compared with control mice. In summary, these studies suggest that muscle LPL is particularly potent with regard to VLDL metabolism and is sufficient to compensate for the lack of LPL in other tissues with regard to lipolyzing VLDL particles. With regard to HDL, muscle LPL expression does not result in normal levels due to enhanced breakdown either by mediating accelerated HDL clearance or by failing to establish normal HDL particles that are then cleared more quickly than normal. These studies provide new insights on the tissue-specific effects of LPL on lipoprotein metabolism. PMID- 9202041 TI - The protein kinase CK2 site (Ser111/112) enhances recognition of the simian virus 40 large T-antigen nuclear localization sequence by importin. AB - The mechanism by which phosphorylation regulates nuclear localization sequence (NLS)-dependent nuclear protein import is largely unclear. Whereas nuclear accumulation of SV40 large tumor antigen (T-ag) fusion proteins is completely dependent on the T-ag NLS (amino acids 126-132), the rate of nuclear import is increased 50-fold by amino acid residues 111-125 and in particular a site for the protein kinase CK2 (CK2) at serine 111/112. Because the first step of nuclear protein import involves the binding of the NLS by an NLS-receptor complex such as the importin 58/97 heterodimer, we established a novel enzyme-linked immunosorbent assay to test whether NLS recognition is influenced by amino acids amino-terminal to the NLS and the CK2 site. We found that recognition of the T-ag NLS by importin 58/97 was enhanced 10-fold in the presence of amino acid residues 111-125 and strongly dependent on importin 97. A T-ag fusion protein in which the spacer between the CK2 site and the NLS was decreased showed 30% reduced binding by importin 58/97. Maximal nuclear accumulation of this protein was reduced by more than 50%, indicating the physiological importance of the correctly positioned CK2 site. Phosphorylation by CK2 increased the T-ag NLS binding affinity for importin 58/97 by a further 40%. We conclude that flanking sequences and in particular phosphorylation at the CK2 site are mechanistically important in NLS recognition and represent the basis of their enhancement of T-ag nuclear import. This study thus represents the first elucidation of the mechanistic basis of the regulation of nuclear protein import through phosphorylation within a phosphorylation-regulated NLS. PMID- 9202042 TI - Phospholipase A2 is necessary for tumor necrosis factor alpha-induced ceramide generation in L929 cells. AB - The role of cytosolic phospholipase A2 (cPLA2) in the regulation of ceramide formation was examined in a cell line (L929) responsive to the cytotoxic action of tumor necrosis factor alpha (TNFalpha). In L929 cells, the addition of TNFalpha resulted in the release of arachidonate, which was followed by a prolonged accumulation of ceramide occurring over 5-12 h and reaching 250% over base line. The formation of ceramide was accompanied by the hydrolysis of sphingomyelin and the activation of three distinct sphingomyelinases (neutral Mg2+-dependent, neutral Mg2+-independent, and acidic enzymes). The variant cell line C12, which lacks cPLA2, is resistant to the cytotoxic action of TNFalpha. TNFalpha was able to activate nuclear factor kappaB in both the wild-type L929 cells and the C12 cells. However, TNFalpha was unable to cause the release of arachidonate or the accumulation of ceramide in C12 cells. C6-ceramide overcame the resistance to TNFalpha and caused cell death in C12 cells to a level similar to that in L929 cells. The introduction of the cPLA2 gene into C12 cells resulted in partial restoration of TNFalpha-induced arachidonate release, ceramide accumulation, and cytotoxicity. This study suggests that cPLA2 is a necessary component in the pathways leading to ceramide accumulation and cell death. PMID- 9202043 TI - Phosphorylation of the respiratory burst oxidase subunit p67(phox) during human neutrophil activation. Regulation by protein kinase C-dependent and independent pathways. AB - The respiratory burst oxidase of phagocytes and B lymphocytes catalyzes the reduction of oxygen to superoxide anion (O-2) at the expense of NADPH. This multicomponent enzyme is dormant in resting cells but is activated on exposure to an appropriate stimulus. The phosphorylation-dependent mechanisms regulating the activation of the respiratory burst oxidase are unclear, particularly the phosphorylation status of the cytosolic component p67(phox). In this study, we found that activation of human neutrophils with formyl-methionyl-leucyl phenylalanine (fMLP), a chemotactic peptide, or phorbol myristate acetate (PMA), a stimulator of protein kinase C (PKC), resulted in the phosphorylation of p67(phox). Using an anti-p67(phox) antibody or an anti-p47(phox) antibody, we showed that phosphorylated p67(phox) and p47(phox) form a complex. Phosphoamino acid analysis of the phosphorylated p67(phox) revealed only 32P-labeled serine residues. Two-dimensional tryptic peptide mapping analysis showed that p67(phox) is phosphorylated at the same peptide whether fMLP or PMA is used as a stimulus. In addition, PKC induced the phosphorylation of recombinant GST-p67(phox) in vitro, at the same peptide as that phosphorylated in intact cells. PMA-induced phosphorylation of p67(phox) was strongly inhibited by the PKC inhibitor GF109203X. In contrast, fMLP-induced phosphorylation was minimally affected by this PKC inhibitor. Taken together, these results show that p67(phox) is phosphorylated in human neutrophils by different pathways, one of which involves protein kinase C. PMID- 9202044 TI - Genetic evidence for a tyrosine kinase cascade preceding the mitogen-activated protein kinase cascade in vertebrate G protein signaling. AB - The signal transduction pathway from heterotrimeric G proteins to the mitogen activated protein kinase (MAPK) cascade is best understood in the yeast mating pheromone response, in which a serine/threonine protein kinase (STE20) serves as the critical linking component. Little is known in metazoans on how G proteins and the MAPK cascade are coupled. Here we provide genetic and biochemical evidence that a tyrosine kinase cascade bridges G proteins and the MAPK pathway in vertebrate cells. Targeted deletion of tyrosine kinase Csk in avian B lymphoma cells blocks the stimulation of MAPK by Gq-, but not Gi-, coupled receptors. In cells deficient in Bruton's tyrosine kinase (Btk), Gi-coupled receptors failed to activate MAPK, while Gq-coupled receptor-mediated stimulation is unaffected. Taken together with our previous data on tyrosine kinases Lyn and Syk, the Gq coupled pathway requires tyrosine kinases Csk, Lyn, and Syk, while the Gi-coupled pathway requires tyrosine kinases Btk and Syk to feed into the MAPK cascade in these cells. The central role of Syk is further strengthened by data showing that Syk can bind to purified Lyn, Csk, or Btk. PMID- 9202045 TI - Urokinase plasminogen activator and gelatinases are associated with membrane vesicles shed by human HT1080 fibrosarcoma cells. AB - Membrane vesicles are shed by tumor cells both in vivo and in vitro. Although their functions are not well understood, it has been proposed that they may play multiple roles in tumor progression. We characterized membrane vesicles from human HT1080 fibrosarcoma cell cultures for the presence of proteinases involved in tumor invasion. By gelatin zymography and Western blotting, these vesicles showed major bands corresponding to the zymogen and active forms of gelatinase B (MMP-9) and gelatinase A (MMP-2) and to the MMP-9. tissue inhibitor of metalloproteinase 1 complex. Both gelatinases appeared to be associated with the vesicle membrane. HT1080 cell vesicles also showed a strong, plasminogen dependent fibrinolytic activity in 125I fibrin assays; this activity was associated with urokinase plasminogen activator, as shown by casein zymography and Western blotting. Urokinase was bound to its high affinity receptor on the vesicle membrane. Addition of plasminogen resulted in activation of the progelatinases associated with the vesicles, indicating a role of the urokinase plasmin system in MMP-2 and MMP-9 activation. We propose that vesicles shed by tumor cells may provide a large membrane surface for the activation of membrane associated proteinases involved in extracellular matrix degradation and tissue invasion. PMID- 9202046 TI - Mechanism of beta-adrenergic receptor desensitization in cardiac hypertrophy is increased beta-adrenergic receptor kinase. AB - Pressure overload cardiac hypertrophy in the mouse was achieved following 7 days of transverse aortic constriction. This was associated with marked beta adrenergic receptor (beta-AR) desensitization in vivo, as determined by a blunted inotropic response to dobutamine. Extracts from hypertrophied hearts had approximately 3-fold increase in cytosolic and membrane G protein-coupled receptor kinase (GRK) activity. Incubation with specific monoclonal antibodies to inhibit different GRK subtypes showed that the increase in activity could be attributed predominately to the beta-adrenergic receptor kinase (betaARK). Although overexpression of a betaARK inhibitor in hearts of transgenic mice did not alter the development of cardiac hypertrophy, the beta-AR desensitization associated with pressure overload hypertrophy was prevented. To determine whether the induction of betaARK occurred because of a generalized response to cellular hypertrophy, betaARK activity was measured in transgenic mice homozygous for oncogenic ras overexpression in the heart. Despite marked cardiac hypertrophy, no difference in betaARK activity was found in these mice overexpressing oncogenic ras compared with controls. Taken together, these data suggest that betaARK is a central molecule involved in alterations of beta-AR signaling in pressure overload hypertrophy. The mechanism for the increase in betaARK activity appears not to be related to the induction of cellular hypertrophy but to possibly be related to neurohumoral activation. PMID- 9202047 TI - Multiple phosphorylation of SacY, a Bacillus subtilis transcriptional antiterminator negatively controlled by the phosphotransferase system. AB - The Bacillus subtilis SacY transcriptional antiterminator is a regulator involved in sucrose-promoted induction of the sacB gene. SacY activity is negatively controlled by enzyme I and HPr, the general energy coupling proteins of the phosphoenolpyruvate:sugar phosphotransferase system (PTS), and by SacX, a membranal protein homologous to SacP, the B. subtilis sucrose-specific PTS permease. Previous studies suggested that the negative control exerted by the PTS on bacterial antiterminators of the SacY family involves phosphoenolpyruvate dependent phosphorylation by the sugar-specific PTS-permeases. However, data reported herein show direct phosphorylation of SacY by HPr(His approximately P) with no requirement for SacX. Experiments were carried out to determine the phosphorylatable residues in SacY. In silico analyses of SacY and its homologues revealed the modular structure of these proteins as well as four conserved histidines within two homologous domains (here designated P1 and P2), present in 14 distinct mRNA- and DNA-binding bacterial transcriptional regulators. Single or multiple substitutions of these histidyl residues were introduced in SacY by site directed mutagenesis, and their effects on phosphorylation and antitermination activity were examined. In vitro phosphorylation experiments showed that SacY was phosphorylated on three of the conserved histidines. Nevertheless, in vivo studies using cells bearing a sacB'-lacZ reporter fusion, as well as SacY mutants lacking the phosphorylatable histidyls, revealed that only His-99 is directly involved in regulation of SacY antitermination activity. PMID- 9202048 TI - Coupling of epidermal growth factor (EGF) with the antiproliferative activity of cAMP induces neuronal differentiation. AB - Nerve growth factor (NGF) functions as a progression factor with both mitogenic and antimitogenic activities. When PC12 cells are treated with NGF, they advance to the G1 stage of the cell cycle before they differentiate. The correlation between cessation of proliferation and differentiation suggests that the antimitotic activity of NGF may be obligatory for differentiation. Although epidermal growth factor- (EGF) and NGF-treated PC12 cells share several common properties, including activation of the mitogen-activated protein (MAP) kinase pathway and induction of immediate early genes, EGF is mitogenic for PC12 cells and does not normally stimulate differentiation. However, combinations of EGF and low levels of cAMP stimulate differentiation even though neither agent alone does (Mark, M. D., Liu, Y., Wong, S. T., Hinds, T. R., and Storm, D.R. (1995) J. Cell Biol. 130, 701-710). Since EGF is mitogenic for PC12 cells and differentiation may not occur until proliferation is inhibited, differentiation caused by cAMP and EGF may be due to the antiproliferative activity of cAMP. To test this hypothesis, we examined the effect of EGF or combinations of EGF and cAMP on PC12 cell proliferation. EGF alone stimulated proliferation of PC12 cells and increased the levels of several cell cycle progression factors including cdk2, cdk4, and cyclin B1. Cyclic AMP inhibited the EGF-stimulated increases in cell cycle progression factors as well as proliferation. Other antiproliferative agents including rapamycin, mimosine, and nitric oxide agonists also synergized with EGF to stimulate differentiation. These data indicate that the coupling of antiproliferative signals with EGF modifies the biological properties of EGF and converts it to a differentiating growth factor. PMID- 9202050 TI - Epithelial cells as sensors for microbial infection. PMID- 9202049 TI - Perspectives series: cell adhesion in vascular biology. Integrin signaling in vascular biology. PMID- 9202051 TI - Kallistatin is a potent new vasodilator. AB - Kallistatin is a serine proteinase inhibitor which binds to tissue kallikrein and inhibits its activity. The aim of this study is to evaluate if kallistatin has a direct effect on the vasculature and on blood pressure homeostasis. We found that an intravenous bolus injection of human kallistatin caused a rapid, potent, and transient reduction of mean arterial blood pressure in anesthetized rats. Infusion of purified kallistatin (0.07-1.42 nmol/kg) into cannulated rat jugular vein produced a 20-85 mmHg reduction of blood pressure in a dose-dependent manner. Hoe 140, a bradykinin B2-receptor antagonist, had no effect on the hypotensive effect of kallistatin yet it abolished the blood pressure-lowering effect of kinin and kallikrein. Relaxation of isolated aortic rings by kallistatin was observed in the presence (ED50 of 3.4 x 10(-9) M) and in the absence of endothelium (ED50 of 10(-9) M). Rat kallikrein-binding protein, but not kinin or kallikrein, induced vascular relaxation of aortic rings. Neither Hoe 140 nor Nomega-nitro--arginine methyl ester, a nitric oxide synthase inhibitor, affected vasorelaxation induced by kallistatin. Kallistatin also caused dose dependent vasodilation of the renal vasculature in the isolated, perfused rat kidney. Specific kallistatin-binding sites were identified in rat aorta by Scatchard plot analysis with a Kd of 0.25+/-0.07 nM and maximal binding capacity of 47.9+/-10.4 fmol/mg protein (mean+/-SEM, n = 3). These results indicate that kallistatin is a potent vasodilator which may function directly through a vascular smooth muscle mechanism independent of an endothelial bradykinin receptor. This study introduces the potential significance of kallistatin in directly regulating blood pressure to reduce hypertension. PMID- 9202052 TI - Stretch-induced VEGF expression in the heart. AB - Vascular endothelial growth factor (VEGF) is an endothelial cell mitogen involved in vascular development and angiogenesis. Recently we have observed increased VEGF expression in the normal myocardium after myocardial infarction in a rat heart. This study was designed to explore the mechanism responsible for this increase in VEGF expression. Induction of myocardial stretch in an isolated perfused Langendorff preparation by inflation of an intraventricular balloon to an end-diastolic load of 35 mmHg for 30 min resulted in a nearly sixfold increase in VEGF message level not only in the chamber subjected to stretch (left ventricle) but also in the unstretched right ventricle, thus raising the possibility of a soluble factor mediating stretch- induced induction of VEGF expression. This was further confirmed by demonstrating that coronary venous effluent collected from the stretched heart and used to perfuse isolated hearts in which no balloon was present was able to induce VEGF expression in these normal hearts. Inhibition of TGF-beta activity using a neutralizing antibody, but not antagonists/inhibitors of endothelin and angiotensin II, eliminated stretch induced increase in VEGF expression. Staurosporine, a protein kinase C inhibitor, also blocked stretch-induced increase of VEGF expression. Measurement of TGF-beta concentration in the perfusate demonstrated increased amounts of the cytokine after myocardial stretch, and addition of TGF-beta protein to the perfusion buffer resulted in increased VEGF expression in control hearts. These results suggest that stretch-induced increase of VEGF expression in the heart is mediated at least in part by TGF-beta. PMID- 9202053 TI - Receptor-mediated cellular entry of nuclear localizing anti-DNA antibodies via myosin 1. AB - A unique subset of anti-DNA antibodies enters living cells, interacts with DNase 1, and inhibits endonuclease activity, before their nuclear localization and subsequent attenuation of apoptosis. We now report that endocytosis of these immunoglobulins is mediated by cell surface binding to brush border myosin (myosin 1). Cellular entry and internalization via this unique receptor provides initial contact for entry and sorting these immunoglobulins to translocate to the nuclear pore and enter the nucleus, interact with DNase 1 within the cytoplasm, or recycle back to the cell surface. This internalization pathway provides clues to the translocation of large proteins across cell membranes and the functional effects of intracellular antibodies on cytopathology. This is the first demonstration that brush border myosin functions as a specific cell surface receptor for internalization of large proteins. PMID- 9202054 TI - Transgenic dissection of HIV genes involved in lymphoid depletion. AB - Transgenic mice carrying an HIV provirus, with selective deletion of all three structural genes, developed extensive lymphoid depletion which was detected not only in the spleen and lymph nodes but also in the thymus. Mice with a high level of HIV gene expression developed acute disease which resulted in premature death, and mice with a low level of viral transcripts developed chronic disease with long-term survival. Neither HIV replication nor the envelope glycoprotein (gp120) was required for T cell depletion. Despite abundant viral gene expression early in life, cell death did not become evident until about the time of full lymphoid maturation, suggesting that thymopoiesis was not affected. The more mature T cells in the peripheral lymphoid organs and in the thymic medulla were less sensitive to the apoptotic process than the immature T cells in the thymic cortex. Gradual depletion of the T cell compartment in the peripheral lymphoid organs was intimately accompanied by the reciprocal expansion of the B cell compartment, resulting in the almost complete replacement of T lymphocytes with B immunoblasts in lymph nodes. Unlike T cells, which showed abundant HIV gene expression, B cells did not. The transgenic approach may help identify the HIV nonstructural gene(s) responsible for immune deficiency and help facilitate dissection of its role in inducing apoptosis. PMID- 9202055 TI - Collagen from the osteogenesis imperfecta mouse model (oim) shows reduced resistance against tensile stress. AB - Osteogenesis imperfecta (OI) is a disease attributable to any of a large number of possible mutations of type I collagen. The disease is clinically characterized in part by highly brittle bone, the cause of this feature being unknown. Recently a mouse model of OI, designated as osteogenesis imperfecta murine (oim), and having a well defined genetic mutation, has been studied and found to contain mineral crystals different in their alignment with respect to collagen and in their size. These observations are consistent with those reported in human OI and the unusual crystal alignment and size undoubtedly contribute to the reduced mechanical properties of OI bone. While the mineral has been investigated, no information is available on the tensile properties of oim collagen. In this study, the mechanical properties of tendon collagen under tension have been examined for homozygous (oim/oim), heterozygous (+/oim), and control (+/+) mice under native wet conditions. The ultimate stress and strain found for oim/oim collagen were only about half the values for control mice. Assuming that prestrained collagen molecules carry most of the tensile load in normal bone while the mineral confers rigidity and compression stability, the reported results suggest that the brittleness of OI bone in the mouse model may be related to a dramatic reduction of the ultimate tensile strain of the collagen. PMID- 9202056 TI - BCR/ABL induces multiple abnormalities of cytoskeletal function. AB - The BCR/ABL oncogene causes human chronic myelogenous leukemia (CML), a myeloproliferative disease characterized by massive expansion of hematopoietic progenitor cells and cells of the granulocyte lineage. When transfected into murine hematopoietic cell lines, BCR/ABL causes cytokine-independence and enhances viability. There is also growing evidence that p210(BCR/ABL) affects cytoskeletal structure. p210(BCR/ABL) binds to actin, and several cytoskeletal proteins are tyrosine phosphorylated by this oncoprotein. Also, at least one aspect of cytoskeletal function is abnormal, in that the affinity of beta1 integrins for fibronectin is altered in CML cells. However, isolated changes in beta1 integrin function would be unlikely to explain the clinical phenotype of CML. We used time-lapse video microscopy to study cell motility and cell morphology on extracellular cell matrix protein-coated surfaces of a series of cell lines before and after transformation by BCR/ABL. BCR/ABL was associated with a striking increase in spontaneous motility, membrane ruffling, formation of long actin extensions (filopodia) and accelerated the rate of protrusion and retraction of pseudopodia on fibronectin-coated surfaces. Also, while untransformed cells were sessile for long periods, BCR/ABL-transformed cells exhibited persistent motility, except for brief periods during cell division. Using cell lines transformed by a temperature-sensitive mutant of BCR/ABL, these kinetic abnormalities of cytoskeletal function were shown to require BCR/ABL tyrosine kinase activity. Similar abnormalities of cytoskeletal function on fibronectin-coated surfaces were observed when hematopoietic progenitor cells purified by CD34 selection from patients with CML were compared with CD34 positive cells from normal individuals. Interestingly, alpha-interferon treatment was found to slowly revert the abnormal motility phenotype of BCR/ABL-transformed cells towards normal. The increase in spontaneous motility and other defects of cytoskeletal function described here will be useful biological markers of the functional effects of BCR/ABL in hematopoietic cells. PMID- 9202057 TI - Plasmin and plasminogen activator inhibitor type 1 promote cellular motility by regulating the interaction between the urokinase receptor and vitronectin. AB - The urokinase receptor (uPAR) coordinates plasmin-mediated cell-surface proteolysis and promotes cellular adhesion via a binding site for vitronectin on uPAR. Because vitronectin also binds plasminogen activator inhibitor type 1 (PAI 1), and plasmin cleavage of vitronectin reduces PAI-1 binding, we explored the effects of plasmin and PAI-1 on the interaction between uPAR and vitronectin. PAI 1 blocked cellular binding of and adhesion to vitronectin by over 80% (IC50 approximately 5 nM), promoted detachment of uPAR-bearing cells from vitronectin, and increased cellular migration on vitronectin. Limited cleavage of vitronectin by plasmin also abolished cellular binding and adhesion and induced cellular detachment. A series of peptides surrounding a plasmin cleavage site (arginine 361) near the carboxy-terminal end of vitronectin were synthesized. Two peptides spanning res 364-380 blocked binding of uPAR to vitronectin (IC50 approximately 8 25 microM) identifying this region as an important site of uPAR-vitronectin interaction. These data illuminate a complex regulatory scheme for uPAR-dependent cellular adhesion to vitronectin: Active urokinase promotes adhesion and also subsequent detachment through activation of plasmin or complex formation with PAI 1. Excess PAI-1 may also promote migration by blocking cellular adhesion and/or promoting detachment, possibly accounting in part for the strong correlation between PAI-1 expression and tumor cell metastasis. PMID- 9202058 TI - CpG motifs in bacterial DNA cause inflammation in the lower respiratory tract. AB - Since unmethylated CpG motifs are more frequent in DNA from bacteria than vertebrates, and the unmethylated CpG motif has recently been reported to have stimulatory effects on lymphocytes, we speculated that bacterial DNA may induce inflammation in the lower respiratory tract through its content of unmethylated CpG motifs. To determine the role of bacterial DNA in lower airway inflammation, we intratracheally instilled prokaryotic and eukaryotic DNA in C3H/HeBFEJ mice and performed whole lung lavage 4 h after the exposure. Heat denatured, single stranded Escherichia coli genomic DNA (0.06 ng endotoxin/microg DNA) was compared to heat denatured, single stranded calf thymus DNA (0.007 endotoxin/microg DNA). 10 microg of bacterial DNA, in comparison to 10 microg of calf thymus DNA, resulted in a fourfold increase in the concentration of cells (P = 0.0002), a fivefold increase in the concentration of neutrophils (P = 0.0002), a 50-fold increase in the concentration of TNF-alpha (P = 0.001), and a fourfold increase in the concentration of both IL-6 (P = 0.0003) and macrophage inflammatory protein-2 (P = 0.0001) in the lavage fluid. Importantly, instillation of 0.60 ng of E. coli LPS resulted in a negligible inflammatory response. To test whether the stimulatory effects of bacterial DNA are due to its unmethylated CpG dinucleotides, we methylated the bacterial DNA and also prepared 20 base pair oligonucleotides with and without CpG motifs. In comparison to instillation of untreated bacterial DNA, methylation of the bacterial DNA resulted in a significant reduction in the concentration of cells and cytokines in the lower respiratory tract. Moreover, oligonucleotides containing embedded unmethylated CpG motifs resulted in inflammation in the lower respiratory tract that was indistinguishable from that observed with untreated bacterial DNA. In contrast, oligonucleotides without the embedded CpG motifs or with embedded but methylated CpG motifs resulted in significantly less inflammation in the lower respiratory tract. The possible relevance of these data to human disease was shown by extracting and analyzing DNA in sputum from patients with cystic fibrosis (CF). Approximately 0.1 to 1% of this sputum DNA was bacterial. Intratracheal instillation of highly purified CF sputum DNA caused acute inflammation similar to that induced by bacterial DNA. These findings suggest that bacterial DNA, and unmethylated CpG motifs in particular, may play an important pathogenic role in inflammatory lung disease. PMID- 9202059 TI - Hepatic origin of cholesteryl oleate in coronary artery atherosclerosis in African green monkeys. Enrichment by dietary monounsaturated fat. AB - Relationships among plasma lipoprotein cholesterol, cholesterol secretion by the isolated, perfused liver, and coronary artery atherosclerosis were examined in African green monkeys fed diets containing cholesterol and 35% of calories as fat enriched in polyunsaturated, monounsaturated, or saturated fatty acids. The livers of animals fed monounsaturated fat had significantly higher cholesteryl ester concentrations (8.5 mg/g wet wt) than the livers of the other diet groups (3.65 and 3.37 mg/g wet wt for saturated and polyunsaturated fat groups, respectively) and this concentration was highly correlated with plasma cholesterol and apoB concentrations in each diet group. Cholesteryl oleate was 58 and 74. 5% of the liver cholesteryl ester in the saturated and monounsaturated fat groups. In each diet group, perfusate cholesteryl ester accumulation rate was highly correlated to liver and plasma cholesterol concentrations, and to plasma LDL cholesteryl ester content. Cholesteryl oleate was 48 and 67% of the cholesteryl esters that accumulated in perfusate in the saturated and monounsaturated fat animals, and this percentage was very highly correlated (r = 0.9) with plasma apoB concentration. Finally, in these two diet groups, liver perfusate cholesteryl ester accumulation rate was well correlated (r >/= 0.8) to coronary artery cholesteryl ester concentration, a measure of the extent of coronary artery atherosclerosis that occurred over the five years of diet induction in these animals. These data define an important role for the liver in the cholesteryl oleate enrichment of the plasma lipoproteins in the saturated and monounsaturated fat groups, and demonstrate strong relationships among hepatic cholesteryl ester concentration, cholesteryl ester secretion, and LDL particle cholesteryl ester content. The high correlation between liver cholesteryl ester secretion and coronary artery atherosclerosis provides the first direct demonstration of the high degree of importance of hepatic cholesteryl ester secretion in the development of this disease process. The remarkable degree of enrichment of cholesteryl oleate in plasma cholesteryl esters of the monounsaturated fat group may account for the relatively high amount of coronary artery atherosclerosis in this group. PMID- 9202060 TI - Nephropathy in human immunodeficiency virus-1 transgenic mice is due to renal transgene expression. AB - HIV-associated nephropathy (HIVAN) is a progressive glomerular and tubular disease that is increasingly common in AIDS patients and one of the leading causes of end stage renal disease in African Americans. A major unresolved issue in the pathogenesis of HIVAN is whether the kidney disease is due to renal cell infection or a "bystander" phenomenon mediated by systemically dysregulated cytokines. To address this issue, we have used two different experimental approaches and an HIV-1 transgenic mouse line that develops a progressive renal disease histologically similar to HIVAN in humans. In the murine model, kidney tissue expresses the transgene and in heterozygous adults, renal disease develops shortly thereafter. We demonstrate by terminal deoxynucleotide transferase mediated dUTP-biotin nick-end labeling assay that similar to the disease in humans, apoptosis of renal tubular epithelial cells is a component of the molecular pathogenesis. To determine whether apoptosis is due to transgene expression or environmental factors, we treated fetal kidney explants (normal and transgenic) with UV light to induce transgene expression. Apoptosis occurred in transgenic but not normal littermates after stimulation of transgene expression. To confirm a direct effect of HIV expression on the production of HIVAN, we transplanted kidneys between normal and transgenic mice. HIVAN developed in transgenic kidneys transplanted into nontransgenic littermates. Normal kidneys remained disease free when transplanted into transgenic littermates. Thus, the renal disease in the murine model is intrinsic to the kidney. Using two different experimental approaches, we demonstrate a direct effect of transgene expression on the development of HIVAN in the mouse. These studies suggest that in humans, a direct effect of HIV-1 expression is likely the essential cause of HIVAN, rather than an indirect effect of cytokine dysregulation. PMID- 9202061 TI - Aggrecan degradation in human cartilage. Evidence for both matrix metalloproteinase and aggrecanase activity in normal, osteoarthritic, and rheumatoid joints. AB - To examine the activity of matrix metalloproteinases (MMPs) and aggrecanase in control and diseased human articular cartilage, metabolic fragments of aggrecan were detected with monospecific antipeptide antibodies. The distribution and quantity of MMP-generated aggrecan G1 fragments terminating in VDIPEN341 were compared with the distribution of aggrecanase-generated G1 fragments terminating in NITEGE373. Both types of G1 fragments were isolated from osteoarthritic cartilage. The sizes were consistent with a single enzymatic cleavage in the interglobular domain region, with no further proteolytic processing of these fragments. Both neoepitopes were also detected by immunohistochemistry in articular cartilage from patients undergoing joint replacement for osteoarthritis (OA), rheumatoid arthritis (RA), and in cartilage from adults with no known joint disease. In control specimens, the staining intensity for both G1 fragments increased with age, with little staining in cartilage from 22-wk-old fetal samples. There was also an increase with age in the extracted amount of MMP generated neoepitope in relation to both aggrecan and collagen content, confirming the immunohistochemical results. After the age of 20-30 yr this relationship remained at a steady state. The staining for the MMP-generated epitope was most marked in control cartilage exhibiting histological signs of damage, whereas intense staining for the aggrecanase-generated fragment was often noted in adult cartilage lacking overt histological damage. Intense staining for both neoepitopes appeared in the more severely fibrillated, superficial region of the tissue. Intense immunostaining for both VDIPEN- and NITEGE- neoepitopes was also detected in joint cartilage from patients with OA or RA. Cartilage in these specimens was significantly more degraded and high levels of staining for both epitopes was always seen in areas with extensive cartilage damage. The levels of extracted VDIPEN neoepitope relative to collagen or aggrecan in both OA and RA samples were similar to those seen in age-matched control specimens. Immunostaining for both types of aggrecan fragments was seen surrounding the cells but also further removed in the interterritorial matrix. In some regions of the tissue, both neoepitopes were found while in others only one was detected. Thus, generation and/or turnover of these specific catabolic aggrecan fragments is not necessarily coordinated. Our results are consistent with the presence in both normal and arthritic joint cartilage of proteolytic activity against aggrecan based on both classical MMPs and "aggrecanase." PMID- 9202062 TI - Liver-directed gene transfer and prolonged expression of three major human ApoE isoforms in ApoE-deficient mice. AB - Apolipoprotein E (apoE) plays a key role in lipoprotein metabolism and may have other important biological functions. In humans, there are three common, naturally occurring isoforms of apoE that are associated with differences in lipid levels and atherosclerosis. However, the direct in vivo effects of the apoE isoforms on lipoprotein metabolism and atherosclerosis are not yet fully understood. To investigate the effect of the apoE isoforms in vivo, we constructed second-generation recombinant adenoviruses encoding each of the apoE isoforms. These recombinant adenoviruses were injected intravenously into apoE deficient mice fed a Western diet (mean baseline cholesterol level 1401 mg/dl) in order to study their effects in the absence of endogenous mouse apoE. Hepatic expression of apoE3 and apoE4 completely normalized the lipoprotein profile; 3 d after injection, mean plasma cholesterol levels were 194 and 217 mg/ dl, respectively, and this effect was maintained for at least 6 wk. Expression of apoE2 had much less effect on lipoprotein levels (mean cholesterol level 752 mg/dl 3 d after injection), despite much higher plasma levels of apoE2 compared with apoE3 and apoE4; by 6 wk after injection the cholesterol levels had returned to baseline levels in the apoE2-expressing mice. Expression of all three isoforms significantly increased HDL cholesterol levels by approximately threefold and was independent of the cholesterol-lowering effect. ApoE transgene expression was substantially prolonged compared with that achieved using a first generation adenovirus and apoE was readily detected in plasma 3 mo after virus injection. These studies demonstrate: (a) prolonged in vivo expression of human apoE isoforms in apoE deficient mice after second-generation recombinant adenovirus mediated somatic gene transfer; and (b) significantly impaired ability of apoE2 in vivo to mediate clearance of remnant lipoproteins in apoE-deficient mice fed a Western diet compared with apoE3 and apoE4. PMID- 9202063 TI - Characterization of protein kinase C beta isoform activation on the gene expression of transforming growth factor-beta, extracellular matrix components, and prostanoids in the glomeruli of diabetic rats. AB - Induction of protein kinase C (PKC) pathway in the vascular tissues by hyperglycemia has been associated with many of the cellular changes observed in the complications of diabetes. Recently, we have reported that the use of a novel, orally effective specific inhibitor of PKC beta isoform (LY333531) normalized many of the early retinal and renal hemodynamics in rat models of diabetes. In the present study, we have characterized a spectrum of biochemical and molecular abnormalities associated with chronic changes induced by glucose or diabetes in the cultured mesangial cells and renal glomeruli that can be prevented by LY333531. Hyperglycemia increased diacylglycerol (DAG) level in cultured mesangial cells exposed to high concentrations of glucose and activated PKC alpha and beta1 isoforms in the renal glomeruli of diabetic rats. The addition of PKC beta selective inhibitor (LY333531) to cultured mesangial cells inhibited activated PKC activities by high glucose without lowering DAG levels and LY333531 given orally in diabetic rats specifically inhibited the activation of PKC beta1 isoform without decreasing PKC alpha isoform activation. Glucose induced increases in arachidonic acid release, prostaglandin E2 production, and inhibition of Na+-K+ ATPase activities in the cultured mesangial cells were completely prevented by the addition of LY333531. Oral feeding of LY333531 prevented the increased mRNA expression of TGF-beta1 and extracellular matrix components such as fibronectin and alpha1(IV) collagen in the glomeruli of diabetic rats in parallel with inhibition of glomerular PKC activity. These results suggest that the activation of PKC, predominately the beta isoform by hyperglycemia in the mesangial cells and glomeruli can partly contribute to early renal dysfunctions by alteration of prostaglandin production and Na+-K+ ATPase activity as well as the chronic pathological changes by the overexpression of TGF beta1 and extracellular matrix components genes. PMID- 9202064 TI - Anti-A2/RA33 autoantibodies are directed to the RNA binding region of the A2 protein of the heterogeneous nuclear ribonucleoprotein complex. Differential epitope recognition in rheumatoid arthritis, systemic lupus erythematosus, and mixed connective tissue disease. AB - The recently described anti-A2/RA33 autoantibodies occur in 20-40% of patients with RA, SLE, and mixed connective tissue disease (MCTD). They are directed to the A2 protein of the heterogeneous nuclear ribonucleoprotein complex (hnRNP-A2), an abundant nuclear protein associated with the spliceosome. The NH2-terminal half of the antigen contains two conserved RNA binding domains whereas its COOH terminal part is extremely glycine-rich. The aim of this study was to characterize the autoepitopes of hnRNP-A2 and to investigate the effects of anti A2/RA33 autoantibodies on possible functions of the antigen. Using bacterially expressed fragments, two major discontinuous epitopes were identified. One containing the complete second RNA binding domain was recognized by the majority of patients with RA and SLE but not by patients with MCTD. The second epitope contained sequences of both RNA binding domains and was preferentially targeted by patients with MCTD. When the RNA binding properties of the antigen were investigated, oligoribonucleotides containing the sequence motif r(UUAG) were found to bind to a site closely adjacent or overlapping with the epitope targeted by autoantibodies from patients with RA and SLE. Moreover, anti-A2/RA33 autoantibodies from patients with RA or SLE, but not from patients with MCTD, inhibited binding of RNA. Thus, anti-A2/RA33 autoantibodies recognize conformation-dependent epitopes located in a functionally important region of the antigen. Furthermore, the specific recognition of an epitope by MCTD patients may be used as another argument in favor of considering MCTD a distinct connective tissue disease. PMID- 9202065 TI - Direct, MHC-dependent presentation of the drug sulfamethoxazole to human alphabeta T cell clones. AB - T cells can recognize small molecular compounds like drugs. It is thought that covalent binding to MHC bound peptides is required for such a hapten stimulation. Sulfamethoxazole, like most drugs, is not chemically reactive per se, but is thought to gain the ability to covalently bind to proteins after intracellular drug metabolism. The purpose of this study was to investigate how sulfamethoxazole is presented in an immunogenic form to sulfamethoxazole-specific T cell clones. The stimulation of four CD4(+) and two CD8(+) sulfamethoxazole specific T cell clones by different antigen-presenting cells (APC) was measured both by proliferation and cytolytic assays. The MHC restriction was evaluated, first, by inhibition using anti-class I and anti-class II mAb, and second, by the degree of sulfamethoxazole-induced stimulation by partially matched APC. Fixation of APC was performed with glutaraldehyde 0.05%. The clones were specific for sulfamethoxazole without cross-reaction to other sulfonamides. The continuous presence of sulfamethoxazole was required during the assay period since pulsing of the APC was not sufficient to induce proliferation or cytotoxicity. Stimulation of clones required the addition of MHC compatible APC. The APC could be fixed without impairing their ability to present sulfamethoxazole. Sulfamethoxazole can be presented in an unstable, but MHC-restricted fashion, which is independent of processing. These features are best explained by a direct, noncovalent binding of sulfamethoxazole to the MHC-peptide complex. PMID- 9202066 TI - Immune cell-derived beta-endorphin. Production, release, and control of inflammatory pain in rats. AB - Localized inflammation of a rat's hindpaw elicits an accumulation of beta endorphin-(END) containing immune cells. We investigated the production, release, and antinociceptive effects of lymphocyte-derived END in relation to cell trafficking. In normal animals, END and proopiomelanocortin mRNA were less abundant in circulating lymphocytes than in those residing in lymph nodes (LN), suggesting that a finite cell population produces END and homes to LN. Inflammation increased proopiomelanocortin mRNA in cells from noninflamed and inflamed LN. However, END content was increased only in inflamed paw tissue and noninflamed LN-immune cells. Accordingly, corticotropin-releasing factor and IL 1beta released significantly more END from noninflamed than from inflamed LN immune cells. This secretion was receptor specific, calcium dependent, and mimicked by potassium, consistent with vesicular release. Finally, both agents, injected into the inflamed paw, induced analgesia which was blocked by the co administration of antiserum against END. Together, these findings suggest that END-producing lymphocytes home to inflamed tissue where they secrete END to reduce pain. Afterwards they migrate to the regional LN, depleted of the peptide. Consistent with this notion, immunofluorescence studies of cell suspensions revealed that END is contained predominantly within memory-type T cells. Thus, the immune system is important for the control of inflammatory pain. This has implications for the understanding of pain in immunosuppressed conditions like cancer or AIDS. PMID- 9202067 TI - Ectopic expression of decorin protein core causes a generalized growth suppression in neoplastic cells of various histogenetic origin and requires endogenous p21, an inhibitor of cyclin-dependent kinases. AB - Decorin belongs to a family of secreted, small, leucine-rich proteoglycans that affect matrix assembly and cellular growth. Ectopic expression of decorin proteoglycan or protein core as a mutated form lacking any glycosaminoglycan side chains induced growth suppression in neoplastic cells of various histogenetic origins, including tumor cells derived from gastrointestinal, genital, skeletal, cutaneous, or bone marrow tissues. Exogenously added recombinant decorin also suppressed overall growth of the parental cell lines. In all stably-transfected clones, growth retardation was specifically associated with induction of the potent cyclin-dependent kinase inhibitor p21, but not p27, and subsequent translocation of p21 protein into the nuclei of decorin-expressing cells. This led to a greater proportion of the cells arrested in G1 phase of the cell cycle. These changes were independent of functional p53 or retinoblastoma protein. De novo expression of decorin in HCT116 human colon carcinoma cells harboring a disrupted p21 gene failed to induce growth suppression, in contrast to the wild type cells in which p21 and growth arrest could be induced. These findings indicate that ectopic production of decorin protein core can retard the growth of a variety of tumor cells and that endogenous p21 is a required downstream effector of this biological axis. PMID- 9202068 TI - Oncostatin M is a proinflammatory mediator. In vivo effects correlate with endothelial cell expression of inflammatory cytokines and adhesion molecules. AB - Oncostatin M is a member of the IL-6 family of cytokines that is primarily known for its effects on cell growth. Endothelial cells have an abundance of receptors for oncostatin M, and may be its primary target. We determined if oncostatin M induces a key endothelial cell function, initiation of the inflammatory response. We found that subcutaneous injection of oncostatin M in mice caused an acute inflammatory reaction. Oncostatin M in vitro stimulated: (a) polymorphonuclear leukocyte (PMN) transmigration through confluent monolayers of primary human endothelial cells; (b) biphasic PMN adhesion through rapid P-selectin expression, and delayed adhesion mediated by E-selectin synthesis; (c) intercellular adhesion molecule-1 and vascular cell adhesion molecule-1 accumulation; and (d) the expression of PMN activators IL-6, epithelial neutrophil activating peptide-78, growth-related cytokine alpha and growth-related cytokine beta without concomitant IL-8 synthesis. The nature of the response to oncostatin M varied with concentration, suggesting high and low affinity oncostatin M receptors independently stimulated specific responses. Immunohistochemistry showed that macrophage-like cells infiltrating human aortic aneurysms expressed oncostatin M, so it is present during a chronic inflammatory reaction. Therefore, oncostatin M, but not other IL-6 family members, fulfills Koch's postulates as an inflammatory mediator. Since its effects on endothelial cells differ significantly from established mediators like TNFalpha, it may uniquely contribute to the inflammatory cycle. PMID- 9202069 TI - Gene recombination in postmitotic cells. Targeted expression of Cre recombinase provokes cardiac-restricted, site-specific rearrangement in adult ventricular muscle in vivo. AB - Mouse models of human disease can be generated by homologous recombination for germline loss-of-function mutations. However, embryonic-lethal phenotypes and systemic, indirect dysfunction can confound the use of knock-outs to elucidate adult pathophysiology. Site-specific recombination using Cre recombinase can circumvent these pitfalls, in principle, enabling temporal and spatial control of gene recombination. However, direct evidence is lacking for the feasibility of Cre-mediated recombination in postmitotic cells. Here, we exploited transgenic mouse technology plus adenoviral gene transfer to achieve Cre-mediated recombination in cardiac muscle. In vitro, Cre driven by cardiac-specific alpha myosin heavy chain (alphaMyHC) sequences elicited recombination selectively at loxP sites in purified cardiac myocytes, but not cardiac fibroblasts. In vivo, this alphaMyHC-Cre transgene elicited recombination in cardiac muscle, but not other organs, as ascertained by PCR analysis and localization of a recombination dependent reporter protein. Adenoviral delivery of Cre in vivo provoked recombination in postmitotic, adult ventricular myocytes. Recombination between loxP sites was not detected in the absence of Cre. These studies demonstrate the feasibility of using Cre-mediated recombination to regulate gene expression in myocardium, with efficient induction of recombination even in terminally differentiated, postmitotic muscle cells. Moreover, delivery of Cre by viral infection provides a simple strategy to control the timing of recombination in myocardium. PMID- 9202070 TI - Susceptibility to atherosclerosis in mice expressing exclusively apolipoprotein B48 or apolipoprotein B100. AB - All classes of lipoproteins considered to be atherogenic contain apo-B100 or apo B48. However, there is a distinct paucity of data regarding whether lipoproteins containing apo-B48 or apo-B100 differ in their intrinsic ability to promote the development of atherosclerosis. To address this issue, we compared the extent of atherosclerosis in three groups of animals: apo-E-deficient mice (apo-B+/+apo-E-/ ) and apo-E-deficient mice that synthesize exclusively either apo-B48 (apo B48/48apo-E-/-) or apo-B100 (apo-B100/100apo-E-/-). Mice (n = 25 in each group) were fed a chow diet for 200 days, and plasma lipid levels were assessed throughout the study. Compared with the levels in apo-B+/+apo-E-/- mice, the total plasma cholesterol levels were higher in the apo-B48/48apo-E-/- mice and were lower in the apo-B100/100apo-E-/- mice. However, the ranges of cholesterol levels in the three groups overlapped. Compared with those in the apo-B+/+apo-E-/ mice, atherosclerotic lesions were more extensive in the apo-B48/48apo-E-/- mice and less extensive in the apo-B100/100apo-E-/- mice. Once again, however, there was overlap among the three groups. The extent of atherosclerosis in each group of mice correlated significantly with plasma cholesterol levels. In mice from different groups that had similar cholesterol levels, the extent of atherosclerosis was quite similar. Thus, susceptibility to atherosclerosis was dependent on total cholesterol levels. Whether mice synthesized apo-B48 or apo B100 did not appear to have an independent effect on susceptibility to atherosclerosis. PMID- 9202071 TI - Possible evidence for endogenous production of a novel galanin-like peptide. AB - Galanin mRNA and peptide are not detectable in normal islets. We studied the effect of galanin antagonists on insulin secretion in the rat beta cell line, RIN5AH, and in perifused rat islets. In RIN cell membranes galanin and its antagonists showed high affinity for 125I-galanin binding sites [Kd: (galanin) 0.03+/-0.01; Ki for galanin antagonists: (C7) 0.12+/- 0.02, (M35) 0.21+/-0.04, and (M40) 0.22+/-0.03 nM, mean+/- SEM, n = 4]. Galanin (1 microM) inhibited glucose-induced insulin release in islets (control 21.2+/-1.5 vs. galanin 4.5+/ 0.2 fmol/islet per min, P < 0.001, n = 6) and RIN5AH cells (control 0.26+/-0.01 vs. galanin 0.15+/-0.02 pmol/10(6) cells per h, P < 0.001, n = 9). In RIN5AH cells, all antagonists blocked the inhibitory effects of galanin and stimulated insulin release in the absence of galanin. C7 and M40 (1 microM) alone significantly stimulated glucose-induced insulin secretion. Purified porcine galanin antibody (GAb) enhanced glucose-induced insulin release from islets (control 100+/- 16.3% vs. GAb 806.1+/-10.4%, P < 0.001, n = 6), and RIN5AH cells (control 100+/-9.6% vs. GAb 149+/-6.8%, P < 0. 01, n = 6). Western blotting of dexamethasone-treated islet extracts using GAb showed a specific band of similar molecular weight to porcine galanin not detected using a rat specific galanin antibody. One possible explanation for these results is the presence of an endogenous galanin-like peptide. PMID- 9202076 TI - Early research into the vitamins: the work of Wilhelm Stepp. PMID- 9202075 TI - Respiratory syncytial virus infection results in airway hyperresponsiveness and enhanced airway sensitization to allergen. AB - Viral respiratory infections can predispose to the development of asthma by mechanisms that are presently undetermined. Using a murine model of respiratory syncytial virus (RSV) infection, acute infection is associated with airway hyperresponsiveness as well as enhanced responses to subsequent sensitization to allergen. We demonstrate that acute viral infection results in increased airway responsiveness to inhaled methacholine and pulmonary neutrophilic and eosinophilic inflammation. This response is associated with predominant production of Th-1-type cytokines in peribronchial lymph node cells in vitro. Mice sensitized to ovalbumin via the airways after RSV infection developed increased airway responsiveness to methacholine and pulmonary eosinophilic and neutrophilic inflammation, associated with the predominant production of Th-2 type cytokines. Treatment of the mice with anti-IL-5 antibody abolished airway hyperresponsiveness and eosinophilic but not neutrophilic inflammation in both acutely infected mice and mice sensitized after infection. We conclude that RSV infection results in airway hyperresponsiveness in the acute phase and leads to changes in immune function that can enhance the effects of airway sensitization to antigen after infection. In both situations, airway hyperresponsiveness is closely associated with pulmonary eosinophilic inflammation. This model provides a means for further analyzing the influence of viral respiratory infections on airway sensitization and the development of altered airway responsiveness. PMID- 9202072 TI - Inhibitors of the proteasome reduce the accelerated proteolysis in atrophying rat skeletal muscles. AB - Several observations have suggested that the enhanced proteolysis and atrophy of skeletal muscle in various pathological states is due primarily to activation of the ubiquitin-proteasome pathway. To test this idea, we investigated whether peptide aldehyde inhibitors of the proteasome, N-acetyl-leucyl-leucyl-norleucinal (LLN), or the more potent CBZ-leucyl-leucyl-leucinal (MG132) suppressed proteolysis in incubated rat skeletal muscles. These agents (e.g., MG132 at 10 microM) inhibited nonlysosomal protein breakdown by up to 50% (P < 0.01), and this effect was rapidly reversed upon removal of the inhibitor. The peptide aldehydes did not alter protein synthesis or amino acid pools, but improved overall protein balance in the muscle. Upon treatment with MG132, ubiquitin conjugated proteins accumulated in the muscle. The inhibition of muscle proteolysis correlated with efficacy against the proteasome, although these agents could also inhibit calpain-dependent proteolysis induced with Ca2+. These inhibitors had much larger effects on proteolysis in atrophying muscles than in controls. In the denervated soleus undergoing atrophy, the increase in ATP dependent proteolysis was reduced 70% by MG132 (P < 0.001). Similarly, the rise in muscle proteolysis induced by administering thyroid hormones was reduced 40 70% by the inhibitors. Finally, in rats made septic by cecal puncture, the increase in muscle proteolysis was completely blocked by MG132. Thus, the enhanced proteolysis in many catabolic states (including denervation, hyperthyroidism, and sepsis) is due to a proteasome-dependent pathway, and inhibition of proteasome function may be a useful approach to reduce muscle wasting. PMID- 9202074 TI - Rho proteins play a critical role in cell migration during the early phase of mucosal restitution. AB - In the intestine, several growth factors stimulate migration of epithelial cells, contributing to the maintenance of tissue integrity. The Ras-like GTPase Rho regulates a signal transduction pathway linking growth factor receptors to the formation of actin stress fibers and focal adhesions, presumed to be important for motility. Using an in vitro wound-induced migration assay, we have examined the role of Rho GTPases in the migration of IEC-6 and Caco-2 cells, and provide evidence that the Rho GTPases play an essential role in the initial phase of mucosal wound healing. Treatment of the cells with Clostridium difficile toxins A and B, inhibitors of the Rho family GTPases inhibited migration in a dose dependent fashion. Microinjection of the inhibitory exchange factor Rho-guanine nucleotide dissociation inhibitor (GDI), or Clostridium botulinum C3 ADP-ribosyl transferase (C3) toxin, a Rho-ADP-ribosylating exoenzyme, potently inhibited migration. Microinjection of RhoT19N, a dominant negative form of RhoA, or in vitro ADP-ribosylated RhoA impaired the ability of cells to migrate. Rho-GDI and C3 exoenzyme also inhibited EGF-induced migration of IEC-6 cells. These results demonstrate that Rho is required for endogenous and EGF-induced migration of small intestinal crypt cells, and that Rho proteins are essential elements of a mechanism by which growth factors induce cell migration to restitute mucosal integrity. PMID- 9202077 TI - Intestinal uptake and biliary excretion of the isoflavone genistein in rats. AB - The intestinal absorption, biliary excretion and metabolism of genistein, a potent and specific protein tyrosine kinase inhibitor that occurs naturally in soy foods, was examined in anesthetized, adult female rats fitted with indwelling biliary cannulas. 4-14C-Genistein, when infused into the duodenum, was rapidly absorbed from the intestine, taken up by the liver and excreted into the bile as its 7-O-beta-glucuronide conjugate. Cumulative recovery of 14C-radioactivity in the bile over a 4-h period was 70-75% of the dose. When genistein was infused into the portal vein, it was also taken up efficiently by the liver, conjugated with glucuronic acid and transported into bile. However, portal blood collected after duodenal infusions of genistein contained mostly genistein 7-O-beta glucuronide, suggesting that in vivo glucuronidation occurred in the intestinal wall rather than the liver. This was confirmed using everted intestinal sac preparations. Reinfusion of genistein 7-O-beta-glucuronide into the duodenum or into the mid small intestine resulted in its reappearance in the bile, albeit more slowly than when genistein was infused. Over a 4-h collection period, the cumulative recovery of 14C-radioactivity in bile was 27 and 70-75% of the administered dose for duodenal and ileal infusions, respectively. These data indicate that genistein is highly bioavailable in rats and because of its enterohepatic circulation may accumulate within the gastrointestinal tract. PMID- 9202078 TI - The positional distribution of dioleoyl-palmitoyl glycerol influences lymph chylomicron transport, composition and size in rats. AB - The effects of 1,3-dioleoyl-2-palmitoyl glycerol (OPO) on lymph chylomicron transport, composition and size in rats were investigated in comparison with 1,2 dioleoyl-3-palmitoyl glycerol (OOP). The OPO and OOP were prepared by enzymatic transesterification reactions. The concentrations of OPO and OOP in the preparations were 65.7 g/100 g, and the composition of fatty acids was similar for each. The OPO preparation contained triacylglycerols with 76.6% of the palmitic acid in the sn-2 position, whereas 100% of the oleic acid was esterified to the sn-2 position in the OOP preparation. Rats were infused with lipid emulsion containing 150 g/L of OPO or OOP via a stomach catheter. Lymph was collected through the mesenteric lymphatic trunk at 1-h intervals for 12 h. Collected lymph chylomicrons were analyzed for triacylglycerol, fatty acids, apolipoprotein A-I and particle size. The maximum transport rates of triacylglycerols in the OPO group were higher than those in the OOP group. The overall absorption of triacylglycerols, palmitic acid and oleic acid in the OPO group was also higher than that in the OOP group. In the chylomicrons, 60-70% of the fatty acids at the sn-2 position of the infused triacylglycerol was transported at the original position. The transport rates of dioleoyl-palmitoyl glycerol in the OPO group were higher than those in the OOP group. The transport rates of apolipoprotein A-I did not differ between groups, whereas the mean diameter of the chylomicrons in the OPO group was larger than that in the OOP group. These results indicate that OPO is absorbed and transported more effectively than OOP. PMID- 9202073 TI - Intestinal epithelial cells use two distinct pathways for HLA class II antigen processing. AB - Intestinal epithelial cells express a low level of HLA class II molecules constitutively, with elevated levels seen in the setting of mucosal inflammation including inflammatory bowel disease. The ability of intestinal epithelial cells to act as antigen presenting cells for alphabeta CD4(+) T lymphocytes was examined through a molecular analysis of the HLA class II antigen processing pathway. We have shown that intestinal epithelial cells contain abundant constitutive levels of the cathepsin proteases proven to function in HLA class II mediated antigen presentation. Activation of these cells by gamma-IFN induced the expression of invariant chain and HLA-DM alphabeta, thus facilitating the formation of compact, SDS-stable HLA- DR alphabeta heterodimers. Using HLA-DR restricted T cells and retroviral mediated gene transfer of HLA-DR alleles into the intestinal epithelial cell lines HT-29 and T84, we demonstrated efficient antigen processing and presentation to CD4(+) T lymphocytes in the presence of the proinflammatory cytokine gamma-IFN. The class II processing pathway and presentation in the presence of gamma-IFN was indistinguishable from that observed with a conventional antigen presenting cell. Antigen processing also occurred in intestinal epithelial cells in the absence of gamma-IFN, and in contrast to that seen after stimulation with gamma-IFN, required high concentration of antigen and was not inhibited by the protease inhibitor leupeptin. These data suggest the use of two distinct pathways of HLA class II antigen processing in enterocytes with differential immunomodulatory properties in the presence or absence of mucosal inflammation. PMID- 9202079 TI - Vitamin A regulates genes involved in hepatic gluconeogenesis in mice: phosphoenolpyruvate carboxykinase, fructose-1,6-bisphosphatase and 6 phosphofructo-2-kinase/fructose-2,6-bisphosphatase. AB - We examined the effects of vitamin A deficiency and all-trans retinoic acid (RA) supplementation on regulation of three important genes in hepatic gluconeogenesis: the genes for phosphoenolpyruvate carboxykinase (PEPCK), fructose-1,6-bisphosphatase (Fru-1,6-P2ase) and 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase (6-PF-2-K/Fru-2,6-P2ase). Mice were made vitamin A deficient in the second generation by initiating a vitamin A-deficient diet on d 10 of gestation. At 7 wk of age, vitamin A-deficient mice were treated with all-trans RA or vehicle alone and killed for RNA analysis. In liver, vitamin A deficiency resulted in PEPCK mRNA levels that were 74% lower and 6-PF-2-K/Fru-2,6-P2ase mRNA levels that were 42% lower than the respective mRNA measured in control mice. The Fru-1,6-P2ase mRNA abundance was not affected by vitamin A deficiency. The decrease in hepatic PEPCK and 6-PF-2-K/Fru-2,6-P2ase mRNA levels was reversed by treatment with all-trans RA within 3 h of administration. In mice fed the control diet, food deprivation for 15 h resulted in PEPCK mRNA levels that were 3.5-fold higher, Fru-1,6-P2ase mRNA levels that were 2-fold higher, and 6-PF-2-K/Fru-2,6 P2ase mRNA levels that were 3.4-fold higher than in fed mice. Vitamin A-deficient mice did not respond to food deprivation with induced PEPCK mRNA levels, whereas 6-PF-2-K/Fru-2,6-P2ase and Fru-1,6-P2ase mRNA levels were induced. The pattern of 6-PF-2-K/Fru-2,6-P2ase mRNA abundance with vitamin A deficiency and food deprivation was complex and different from that for either PEPCK or Fru-1,6-P2ase transcripts. The cAMP-responsiveness of the PEPCK gene in vitamin A-deficient mice was tested. Vitamin A deficiency caused a significant reduction in cAMP stimulation of PEPCK mRNA levels in liver. These results in the whole animal indicate that vitamin A regulation of the hepatic PEPCK gene is physiologically important; without adequate vitamin A nutriture, stimulation of the PEPCK gene by food deprivation or cAMP treatment is inhibited in the liver. PMID- 9202080 TI - Low dietary protein impairs blood coagulation in BHE/cdb rats. AB - The influence of dietary protein on blood coagulation tests was evaluated in BHE/cdb rats. Three experiments were conducted in order to compare effects of diets with low (8 g/100 g diet) or high (38 g/100 g diet) protein, to establish values for coagulation tests at intermediate (12-30 g/100 g diet) concentrations of dietary protein, and to compare feeding identical quantities of diets with 8 g protein/100 g diet vs. 18 g protein/100 g diet. After 4 wk of feeding the semipurified diets, bleeding time exceeded 15 min in the groups fed low protein diets, compared to a range of 3-6 min for the groups fed high protein diets. Several in vitro tests of coagulation were abnormal in the rats fed low protein diets. For example, prothrombin time averaged 27 +/- 8 s in rats fed 8 g protein/100 g diet plus beef tallow, but 17 +/- 1 s in rats fed 38 g protein/100 g diet plus tallow. The coagulation deficit in rats fed low protein was not affected by fat source (tallow vs. menhaden oil), but fibrinogen was elevated in rats fed diets with menhaden oil. Conversely, no differences in coagulation tests were observed among rats fed 12-30 g protein/100 g diet. Bleeding times ranged from 7 to 9 min, and prothrombin time was 17-18 s. Significant differences in plasma fibrinogen concentration and prothrombin time were observed in rats fed 8 vs. 18 g protein/100 g diet at a fixed rate of 6 g/100 g body weight. Platelet and blood cell numbers were unaffected by dietary protein. The evidence for multiple deficits in the coagulation system suggests that hepatic function in BHE/cdb rats may become impaired when the rats are fed low protein diets of the composition used here. PMID- 9202081 TI - Colostrum enhances the nutritional stimulation of vital organ protein synthesis in neonatal pigs. AB - Our objective was to determine the relative importance of the macronutrient components of colostrum in the stimulation of vital organ protein synthesis in neonatal pigs. We studied colostrum-deprived newborn pigs within 4-6 h after birth (unfed) and three groups fed for 24 h mature milk, colostrum, or a formula containing a macronutrient composition comparable to that of colostrum. We measured protein synthesis in vivo using a flooding dose of 3H-phenylalanine. The fractional rates of protein synthesis (Ks) in the brain, heart, lung, kidney and spleen were significantly higher in all fed groups than in the unfed newborns. Among the three fed groups, brain and heart protein synthesis rates were greater in colostrum-fed than in either milk- or formula-fed pigs. Kidney and spleen protein synthesis rates in colostrum- and formula-fed pigs were not significantly different, but both were higher than in milk-fed pigs. The stimulation of kidney protein synthesis in response to feeding was primarily a consequence of greater protein synthetic efficiency; however, protein synthetic capacity in the heart, lung and spleen was generally greater in colostrum- and formula-fed pigs than in unfed newborns. Our results suggest that the predominant stimulus for vital organ protein synthesis in colostrum-fed neonatal pigs is nutrient intake. However, there was a specific stimulation of both brain and heart protein synthesis in colostrum-fed pigs that cannot be attributed to macronutrients. PMID- 9202082 TI - Supplementation with vitamin C, vitamin E or beta-carotene influences osmotic fragility and oxidative damage of erythrocytes of zinc-deficient rats. AB - Dietary zinc deficiency in rats causes increased osmotic fragility of their erythrocytes. In this study, the influence of supplementary antioxidants (vitamin C, vitamin E or beta-carotene) on osmotic fragility, oxidative damage and components of the primary defense system of erythrocytes of zinc-deficient rats was investigated. Indicators of hemolysis in vivo were also examined. Five groups of 12 male rats were force-fed a zinc-adequate diet (control rats), a zinc deficient diet or a zinc-deficient diet enriched with vitamin C, vitamin E or beta-carotene. Compared with the control rats, the rats fed the zinc-deficient diet without supplementary antioxidants had greater red blood cell osmotic fragility, higher concentrations of thiobarbituric acid-reactive substances and alanine, higher glutathione S-transferase activity, lower concentration of glutathione and activity of glutathione peroxidase as well as lower activity of superoxide dismutase in plasma (P < 0.05). Supplementation with antioxidants generally improved osmotic fragility in zinc-deficient rats without influencing zinc concentration or alkaline phosphatase activity in plasma, indicators of zinc status. At some of the hypotonic saline concentrations tested, vitamin C and beta carotene significantly affected osmotic fragility. The zinc-deficient rats fed a diet without supplementary antioxidants had significantly higher concentrations of alanine in erythrocytes than the zinc-deficient rats supplemented with vitamin C, vitamin E or beta-carotene and had significantly higher levels of thiobarbituric acid-reactive substances in erythrocytes than the rats supplemented with beta-carotene. There was no indication of hemolysis in vivo in rats fed zinc-deficient diets. The results show that supplementary antioxidants decrease osmotic fragility and oxidative damage of erythrocytes in zinc-deficient rats. PMID- 9202084 TI - Selenium regulation of classical glutathione peroxidase expression requires the 3' untranslated region in Chinese hamster ovary cells. AB - Classical glutathione peroxidase (GPX) mRNA levels fall dramatically in selenium (Se)-deficient animals, but it is not known whether this mechanism is related to the mRNA 3' untranslated region (3'UTR) sequences that have been shown to direct Se incorporation. In this study, we used recombinant GPX constructs to investigate the role of the GPX 3'UTR in Se regulation of GPX mRNA levels in Chinese hamster ovary (CHO) cells. The CHO cells were transfected with GPX (pRc/GPX), GPX lacking the 3'UTR (pRc/Delta3'UTR) or the pRc/CMV vector alone, and GPX activity and GPX mRNA levels were determined in stable transfectants grown in low Se basal medium with a range of added Se concentrations. We identified two pRc/GPX transfectants with significantly elevated GPX activity levels compared with pRc/CMV transfectants. The elevated GPX expression did not dramatically shift the amount of Se that was sufficient for GPX activity to reach the Se-adequate plateau level (100 nmol/L added Se). As expected, GPX activity was not significantly different when pRc/Delta3'UTR transfectants were compared with pRc/CMV control transfectants. Among the wild type and transfected CHO cells, Se-deficient GPX activity levels averaged 35 +/- 5% of Se-adequate levels. Selenium-deficient levels of endogenous GPX mRNA as well as recombinant pRc/GPX mRNA averaged 54-58% of Se-adequate levels; 3-4 nmol/L added Se was sufficient for maximal GPX mRNA levels. In contrast, pRc/Delta3'UTR mRNA levels in the unsupplemented cells remained at Se-adequate levels and showed no distinct Se regulation. These studies demonstrate that the GPX 3'UTR is necessary for Se regulation of GPX mRNA levels in addition to its role in Se incorporation. PMID- 9202083 TI - Retinoic acid stimulates early cellular proliferation in the adapting remnant rat small intestine after partial resection. AB - Following loss of small bowel surface area, the remnant intestine undergoes a remarkable adaptive response. To define more fully the underlying molecular mechanisms, we have identified genes that are specifically induced in the adapting remnant after partial small bowel resection. Several of these, including cellular retinol binding protein II (CRBP II) and apolipoprotein (apo) AI, participate in vitamin A and lipid trafficking. The CRBP II and apo A-I promoters contain response elements for the nuclear retinoid X receptor RXR-alpha. It is well established that vitamin A is essential for normal cell growth, differentiation and maintenance of epithelial tissues and that CRBP II functions to facilitate intestinal vitamin A absorption and metabolism. On the basis of these considerations, changes in CRBP II and apo A-I mRNA levels could reflect a role for retinoids in modulating the intestinal adaptive response. To explore this hypothesis, we used a rat resection model of intestinal adaptation to examine the temporal patterns of CRBP II, apo A-I and RXR-alpha expression postresection. CRBP II and apo A-I mRNA levels were increased in the remnant intestine in distinct temporal patterns, whereas RXR-alpha expression was unchanged. To address directly the effects of vitamin A in adaptation, retinoic acid or vehicle was administered intravenously to rats immediately after 70% small bowel resection. Compared with vehicle, all-trans-retinoic acid significantly stimulated crypt cell proliferation in the adapting remnant intestine by 6 h after surgery. These data suggest that retinoic acid acts to modulate intestinal proliferation in the adapting small intestine after loss of functional small bowel surface area. PMID- 9202085 TI - Dietary triacylglycerols with palmitic acid (16:0) in the 2-position increase 16:0 in the 2-position of plasma and chylomicron triacylglycerols, but reduce phospholipid arachidonic and docosahexaenoic acids, and alter cholesteryl ester metabolism in formula-Fed piglets. AB - Milk triacylglycerols have an unusual fatty acid distribution, with palmitic acid (16:0) esterified predominately at the center (sn-2) position. Other dietary triacylglycerols contain 16:0 predominantly at the sn-1,3 positions. This study was designed to evaluate the effect of formula triacylglycerol fatty acid distribution on the composition and distribution of plasma lipoprotein fatty acids in piglets fed formula containing synthesized triacylglycerols or palm olein oil with about 32 or 4.2% 16:0, respectively, in fatty acids at the sn-2 position, with comparison to piglets fed sow's milk. Feeding formula with 16:0 at the triglyceride sn-2 position or sow's milk resulted in higher chylomicron triacylglycerol sn-2 16:0 than when palm olein was fed. This suggests that dietary triacylglycerol sn-2 position fatty acids are conserved during digestion, absorption and reassembly to chylomicron triacylglycerols. The increased chylomicron triacylglycerol sn-2 position 16:0 in piglets fed synthesized triacylglycerols was accompanied by lower chylomicron triacylglycerol arachidonic and docosahexaenoic acid than in piglets fed formula with palm olein, suggesting an interaction between dietary triacylglycerol saturated fatty acid distribution and (n-6) and (n-3) fatty acid transport. PMID- 9202086 TI - Von Willebrand factor restores impaired platelet thrombogenesis in copper deficient rats. AB - Dietary copper restriction reduces microvascular thrombogenesis. We have now examined the roles of shear forces and von Willebrand factor (vWF) in in vivo thrombus formation in the cremaster microcirculation of copper-deficient rats. Male weanling Sprague-Dawley rats were fed purified diets that were either copper adequate (6.3 mg Cu/kg) or copper-deficient (0.3 mg Cu/kg) for 4 wk. Intravascular fluorescein isothiocyanate tagged to bovine serum albumin was activated with 450-490 nm light to induce thrombus formation in microvessels. Thrombus initiation time was significantly prolonged in copper-deficient rats; after thrombus appearance, however, vessel occlusion was significantly accelerated. The greater shear rates of arterioles compared with venules significantly increased the thrombus initiation time in both groups. However, vessel occlusion time and thrombus growth time were independent of shear rate. Intravascular vWF (0.2 u/100 g body wt) decreased thrombus initiation time in the CuD group without affecting thrombus growth time. The data suggest that decreased thrombogenesis in copper-deficient rats is not a result of altered rheological factors or arteriolar-venular differences, but appears to result from decreased platelet-to-endothelial cell adhesion. PMID- 9202089 TI - Genetic variation in cholesterol absorption efficiency among inbred strains of mice. AB - The initial study utilized the outbred Black Swiss, the inbred 129/SvEv and their hybrid mice to test for possible genetic difference in cholesterol absorption efficiency. Female mice (10-12 wk old) were fed a lipid test meal containing [3H]cholesterol and beta-[14C]sitosterol by stomach tube. The amount of [3H]cholesterol excreted in the feces was determined as nonabsorbed cholesterol and was normalized based on the recovery of the nonabsorbable beta [14C]sitosterol. The Black Swiss mice absorbed significantly less cholesterol than the 129/SvEv mice within a 24-h period. Cholesterol absorption efficiency of the hybrid mice varied widely and did not segregate with either parental group. Differences in cholesterol absorption efficiency were also observed among six different inbred strains of mice fed either a basal low fat diet or a high fat/high cholesterol diet for 3 wk. Cholesterol absorption efficiency did not differ among DBA/2, C57BL/6, C3H/He, BALB/c and AKR/J mice under basal dietary conditions. However, cholesterol absorption was significantly lower in the DBA/2 mice than in C57BL/6 and C3H/He mice after mice were fed a high fat/high cholesterol diet. Cholesterol absorption by the C57L/J mice did not differ from that of C57BL/6, C3H/He, BALB/c and AKR/J mice under basal diet conditions, but was significantly lower when mice were fed a high fat/high cholesterol diet. Cholesterol absorption efficiency differed between DBA/2 and C57L/J mice under both dietary conditions. These results suggest that cholesterol absorption is controlled by multiple genetic factors. PMID- 9202088 TI - Toxic damage to the respiratory epithelium induces acute phase changes in vitamin A metabolism without depleting retinol stores of South African children. AB - Whereas there is much information concerning the effects of vitamin A status on response to infectious challenge, the effects of infection or trauma on vitamin A metabolism and status are less well documented. These relationships need to be understood to optimize clinical and public health programs to improve vitamin A status and health of children in less-developed countries. We measured acute changes in retinol and retinol-binding protein in 57 young South African children hospitalized following respiratory epithelial damage resulting from accidental ingestion of kerosene. In addition, vitamin A status, as measured by the modified relative dose response test, of these children 3 mo later was compared with that of neighborhood control children to determine whether their illness had depleted retinol stores. Plasma retinol was already significantly below control levels when children were admitted [geometric mean (95% CI): 0.57 micromol/L (0.48-0.67) compared with 1.15 micromol/L (1.02-1.30) for controls] and decreased further the following morning [0.38 micromol/L (0.31-0.46)]. Significant differences in retinol-binding protein were not detected until the next morning [5.99 mg/L (4.70 7.63) compared with 14.0 mg/L (11.8-16.6) for controls] and were not as large as the relative differences in retinol. This dissociation between changes in retinol and its binding protein suggests that there may be increased retinol uptake by certain tissues during the acute phase response. The proportion of case children (37/46, 80%) with inadequate liver retinol stores 3 mo after the illness was slightly, but not significantly (chi2 = 2.16, P = 0.14), greater than the proportion of control children (28/42, 67%). Acute respiratory illness therefore did not further deplete retinol stores in this population in which stores were already frequently inadequate. PMID- 9202087 TI - Zinc supplementation affects the activity patterns of rural Guatemalan infants. AB - Zinc deficiency has been associated with growth deficits, reduced dietary intake and appetite, and has been hypothesized to result in reduced activity. This randomized, double-blind, placebo-controlled study examined whether 10 mg of oral zinc as zinc sulfate, given daily for up to 7 mo, affected activity patterns of 85 Guatemalan infants recruited at 6-9 mo of age. Infant activity was assessed by time sampling-observation method at 10-min intervals during a 12-h data collection period, at base line, 3 and 7 mo follow-up. Motor development and the percentage of time infants were observed in various positions (being carried, lying down, sitting, crawling, standing or walking) and engaged in various activities (eating, sleeping, resting, crying/whining or playing) were compared by treatment group. No differences in motor development were observed by treatment group. However, at follow-up 2 (after 7 mo of supplementation), zinc supplemented infants were significantly more frequently observed sitting up compared with lying down, and were playing during 4.18 +/- 1.95% (P < 0.05) more observations than unsupplemented infants. They were also somewhat less likely to be observed crying or whining (P < 0.10) compared with those receiving the placebo. These effects are independent of other factors including infant age, motor development, sex, maternal education, family socioeconomic status and nutritional status at base line. Further research must be conducted to determine the long-term developmental importance of these differences in activity patterns associated with zinc supplementation in this setting. PMID- 9202090 TI - Soluble amylose cornstarch is more digestible than soluble amylopectin potato starch in rats. AB - In liquid enteral formulations, high molecular weight soluble starches may be able to replace glucose and low molecular weight glucose polymers that have high glycemic indices. Male rats were fed either commercial cornstarch, dextrose, modified soluble potato (70-75% amylopectin) starch, or modified soluble amylomaize-7 (70% amylose) starch for 4 wk. Body weights did not differ among the groups. Food consumption was significantly higher in the two modified starch-fed groups than in the two control groups. Commercial cornstarch, dextrose, modified potato starch and modified amylomaize-7 starch were 100 +/- 0, 100 +/- 0, 69.0 +/ 1.0 and 91.5 +/- 0.8% digestible, respectively (n = 9, mean +/- SEM). The modified potato starch-fed group deposited the least fat, protein and energy. In both modified starch-fed groups, liver weights were significantly greater than in the two control groups. In food-deprived rats, serum free fatty acid concentrations in the modified potato starch-fed group were significantly higher than in the two control groups, and serum glucose concentrations were significantly higher in the two modified starch-fed groups than in the controls. The insulin to glucagon ratios were significantly lower in the modified potato starch-fed and amylomaize-7 starch-fed groups than in the dextrose-fed control group. Serum protein concentrations, measured after food deprivation, were significantly lower in the modified potato starch-fed group than in the other three groups. Gluconeogenesis from fermentation products might account for the high serum glucose concentrations in the two experimental groups. These data indicate that only the modified amylomaize-7 starch may be useful in the development of food products for liquid nutritional supplements because of the high digestibility and the low resultant insulin levels. PMID- 9202091 TI - Dietary skim milk powder increases ionized calcium in the small intestine of piglets compared to dietary defatted soybean flour. AB - Calcium distribution was studied in the small intestine of piglets fed skim milk powder (SMP) or defatted soybean flour (DSF ) to investigate the relationship between calcium availability and its forms. Ionized calcium in duodenal and ileal digesta was measured with a selective calcium electrode that was not affected by changes in pH or sodium, potassium and magnesium concentrations, which simulated the liquid phases of digesta. Eight piglets were fed DSF-based diet or SMP-based diet for 30 d, and duodenal and ileal digesta were collected. Soluble calcium concentrations in the ileum were higher in the SMP-fed group than in the DSF-fed group. The proportion of soluble calcium in higher-molecular-weight fraction (MW > 3000) was significantly greater in the ileum than in the duodenum of the SMP group, but did not differ between these intestinal segments within the DSF group. This proportion was significantly higher in the ileum of the SMP-fed group than in that of the DSF-fed group. In the ileum, ionized calcium concentration was significantly greater in the SMP-fed group than in the DSF-fed group. These results suggest that the increase of calcium in the higher-molecular-weight fraction raises soluble calcium concentration and changes the distribution of calcium in the ileum of the SMP-fed group. The complexes of calcium with higher molecular-weight ligands may be easily exchangeable with ionized calcium, and the increase in these calcium complexes may consequently enhance the recruitment of ionized calcium, which then can be absorbed. PMID- 9202092 TI - Formula containing randomized fats with palmitic acid (16:0) in the 2-position increases 16:0 in the 2-position of plasma and chylomicron triglycerides in formula-fed piglets to levels approaching those of piglets fed sow's milk. AB - Human and pig milk fat contains a high proportion of palmitic acid (16:0) which is largely esterified to the 2-position of the triglycerides. In contrast, the 16:0 in most nonmilk fats and in infant formulas is mainly esterified at the triglyceride 1,3 positions. Gastric and pancreatic lipases hydrolyze fatty acids from the dietary triglyceride 1- and 3-positions to produce unesterified fatty acids and 2-monoglycerides which are absorbed and re-esterified. In this study, we determined whether formula with chemically randomized oils, which equally distributes 16:0 among all the positions of triglycerides, influences growth or the distribution of fatty acids in plasma and liver lipid of formula-fed piglets compared with piglets fed formula with native oils or sow's milk. After feeding from birth to 18 d, piglets fed formula with palm olein randomized with canola oil (co-randomized) had higher weight gain per liter of formula intake and higher 16:0 in the chylomicron triglyceride 2-position than piglets fed formula with randomized or native palm olein oil blended with canola oil. The fatty acid distribution of formula triglycerides is an important determinant of pathways of 16:0 absorption, and consequently of plasma lipid fatty acids in formula-fed piglets. PMID- 9202093 TI - Maternal semistarvation and streptozotocin-diabetes in rats have different effects on the in vivo glucose uptake by peripheral tissues in their female adult offspring. AB - Previous work in humans and rats has revealed a link between perinatal growth retardation and glucose intolerance in adulthood. Both maternal semistarvation and severe diabetes are accompanied by perinatal growth retardation in rats. In this study, we compared the effect of these conditions on tissue glucose uptake in their female offspring. Glucose uptake was measured as glucose metabolic index (GMI), using 2-deoxy-[1-3H]-glucose, in the postabsorptive state and during euglycemic hyperinsulinemia. The GMI was measured in insulin-sensitive tissues (5 skeletal muscles, diaphragm and white adipose tissue) and in two noninsulin sensitive tissues (duodenum and brain) of adult offspring of normal dams, dams rendered diabetic with streptozotocin on d 11 of pregnancy, and dams fed half normal rations from d 11 of pregnancy. Whole-body insulin resistance, measured by decreased glucose infusion rate during hyperinsulinemia, was milder in offspring of semistarved rats (O-SR) than in offspring of diabetic rats (O-DR). The basal GMI did not differ among the three groups in any tissue except tibialis anterior; during hyperinsulinemia, GMI was significantly greater in the insulin-sensitive tissues of all three groups. GMI of skeletal muscles and adipose tissue during hyperinsulinemia did not differ between control rats and O-SR; in contrast, the GMI was 25-50% lower in skeletal muscles of O-DR during hyperinsulinemia than in those of control rats or O-SR. Thus, maternal semistarvation and diabetes have dissimilar effects on peripheral insulin sensitivity of the adult female offspring. Because both conditions are associated with perinatal growth retardation and fetal hypoinsulinemia, other mechanisms must be identified to explain impaired glucose uptake by skeletal muscles in the offspring of diabetic rats. PMID- 9202094 TI - Long-term consumption of an amino acid diet reduces the pancreatic enzyme secretion response to a trypsin inhibitor in rats. AB - We investigated pancreatic enzyme secretion in response to soybean trypsin inhibitor (SBTI) in rats fed amino acids as a nitrogen source, from the fetal stage to adulthood. Pregnant rats were divided into two groups 4 d before parturition. During gestation and nursing, one group was fed a 15% protein diet (protein-fed rats) and the other (amino acid-fed rats) a 15% amino acid mixture diet that simulated the composition of the protein diet. Each male offspring was weaned at 4 wk after parturition and fed the same diet as fed to its dam for an additional 6 wk. Pancreatic amylase secretion in response to an intraduodenal infusion of SBTI (10 mg/rat) was observed in the protein-fed rats but not in the amino acid-fed rats. Amylase secretion in response to an intravenous injection of cholecystokinin (CCK) (10 ng/kg rat) was observed in both groups, and the magnitude of the response was significantly higher in the amino acid-fed rats than in the protein-fed rats. An increase in the level of plasma CCK in response to SBTI was observed in the protein-fed rats but not in the amino acid-fed rats. These results suggest that the long-term amino acid diet, because of its ability to inhibit the SBTI-stimulated CCK-releasing process in the small intestine of rats, reduced the pancreatic enzyme secretion response to a trypsin inhibitor. Six rats fed the amino acid mixture until 1 wk after weaning were fed the protein diet for the next 5 wk. These rats showed no pancreatic amylase secretion in response to SBTI, suggesting that dietary components around the weaning stage may affect the development of the ability of small intestinal cells to recognize a trypsin inhibitor. PMID- 9202095 TI - Total intestinal lactase and sucrase activities are reduced in aged rats. AB - Lactase-phlorizin hydrolase (LPH) and sucrase-isomaltase (SI) are intestinal microvillus membrane hydrolases that play important roles in carbohydrate digestion. Although the expression of these enzymes during postnatal development has been characterized, the effect of old age on disaccharidase activity is poorly understood. In the present investigation, we examined the effect of aging on lactase and sucrase activities and their mRNA levels in the small intestines of 3-, 12- and 24- mo-old rats by sampling from nine equidistant segments of small intestine. Total intestinal disaccharidase activity or mRNA abundance was determined from areas under the proximal-to-distal curves. Rats 24 mo of age had total intestinal lactase and sucrase activities that were 12 and 38% lower, respectively, than the 3-mo-old animals (P < 0.05). In contrast, total LPH and SI mRNA abundance did not change significantly. Thus, total intestinal lactase and sucrase activities decrease with age in a manner that likely involves a posttranscriptional process. The age-related decline in disaccharidase activity, if extrapolated to humans, may have important implications for the digestion of carbohydrate contained in the diet of the elderly. PMID- 9202096 TI - Fish oil source differentially affects rat immune cell alpha-tocopherol concentration. AB - We have previously reported that both the source of dietary fish oil and the chemical form of vitamin E supplied in the diet affect the vitamin E status of immune cells in rats. The purpose of this study was to investigate further the effect of fish oil source on immune cell vitamin E status using free alpha tocopherol (alpha-T) at the AIN recommended level as the sole source of vitamin E. Sixty weanling female rats were fed semipurified, high fat (20 g/100 g) diets containing either tocopherol-stripped lard (LRD), menhaden fish oil (MFO), sardine fish oil (SRD) or cod liver oil (CLO) as the primary lipid source. Endogenous alpha-T concentration was measured and equalized to 150 mg/kg oil by addition of free RRR-alpha-T to each lipid source, allowing for a final concentration of alpha-T in the mixed diet of 30 mg/kg. An additional group of rats was fed LRD without supplemental vitamin E (LRD-) as a negative control. After feeding experimental diets for 5 or 10 wk, tissues were collected for alpha T analysis by HPLC. After 5 wk, plasma and liver alpha-T (micromol alpha-T/g lipid) were significantly lower in SRD- and CLO-fed rats compared with LRD-fed rats. At 10 wk, only plasma alpha-T in CLO-fed rats remained significantly depressed. Plasma and liver alpha-T concentrations (micromol alpha-T/g lipid) were not significantly lower in MFO-fed rats than LRD-fed rats at either time point. Compared with LRD, feeding MFO to rats for 5 or 10 wk resulted in significantly greater alpha-T content of immune cells. In similar fashion, SRD fed rats, compared with LRD-fed rats, also had significantly greater alpha-T content in splenocytes at both time points and greater thymocyte alpha-T at 10 wk. In all instances, the alpha-T status of rats fed CLO was indistinguishable from that of rats fed the vitamin E-free diet (LRD-). These data further demonstrate the complexity of the relationship between vitamin E status and dietary (n-3) polyunsaturated fatty acids (PUFA). PMID- 9202097 TI - Consumption of buckwheat protein lowers plasma cholesterol and raises fecal neutral sterols in cholesterol-Fed rats because of its low digestibility. AB - Buckwheat protein product (BWP) has a strong hypocholesterolemic activity in rats fed a cholesterol-enriched diet. In this study, we examined the influence of BWP on fecal excretion of sterols and nitrogen in rats fed a diet containing 5 g/kg cholesterol and 1.25 g/kg sodium cholate, and we examined whether the cholesterol lowering activity of BWP is due to its low digestibility. In Experiment 1, rats fed BWP for 3 wk had significantly lower concentrations of plasma cholesterol and enhanced excretion of fecal total neutral sterols and nitrogen compared with rats fed casein. There was a significant correlation between fecal total neutral sterols and nitrogen (r = 0.89, P < 0.01). Fecal excretion of acidic sterols was unaffected by BWP. In Experiment 2, plasma cholesterol in rats fed trypsin digested BWP for 2 wk was significantly higher than that in rats fed intact BWP. In Experiment 3, rats were fed BWP, low-molecular-weight fraction of the digest of BWP (LMF ) or high-molecular-weight fraction of the digest of BWP (HMF ) for 3 wk. Plasma cholesterol was lower in the BWP group than in the LMF group (P < 0.05), whereas that in the HMF group was intermediate. The in vitro digestibility of BWP with pepsin and pancreatin was significantly lower than that of casein. The results suggest that the cholesterol-lowering effect of BWP is mediated by higher fecal excretion of neutral sterols and that lower digestibility of BWP is at least partially responsible for the effect. PMID- 9202099 TI - Milk inhibits and ascorbic acid favors ferrous bis-glycine chelate bioavailability in humans. AB - The chemical properties of ferrous bis-glycine chelate allow for its use as a fortificant in fluid, high fat vehicles. This chemical form may also protect iron from the inhibitory or enhancing effects of the diet on iron absorption. Alternatively, iron bis-glycine chelate may be absorbed by a mechanism independent of an individual's iron stores. To test these hypotheses, the bioavailability of iron bis-glycine chelate added to water and milk was studied using a double-isotopic method in two groups of 14 women. Iron absorption from aqueous solutions of 0.27 mmol/L (15 mg/L) of elemental iron as either iron bis glycine or ferrous ascorbate was not significantly different (34.6 and 29.9%, respectively). There were significant correlations between (log) iron absorption of iron bis-glycine with (log) serum ferritin (r = -0.60, P < 0.03) and with (log) iron absorption from ferrous ascorbate (r = 0.71, P < 0.006), suggesting that iron bis-glycine chelate bioavailability is indeed affected by iron stores. Iron absorption of iron bis-glycine given in milk was significantly lower (P < 0.002) than when given in water, with values of 11.1 and 46.3%, respectively (standardized to 40% absorption of the reference dose). With the addition of 0.57 mmol/L ascorbic acid (100 mg/L), iron absorption of iron bis-glycine given in milk increased significantly from 11.1 to 15.4% (P < 0.05). These findings show that milk and ascorbic acid affect iron bis-glycine chelate bioavailability and also demonstrate that iron stores may influence its bioavailability as well. The good bioavailability of iron bis-glycine makes this compound a suitable alternative to be considered in iron fortification programs. PMID- 9202098 TI - Oxidative stress and antioxidant status in mouse liver: effects of dietary lipid, vitamin E and iron. AB - The purpose of this study was to determine the effects of dietary fat, vitamin E and iron on oxidative damage and antioxidant status. Male Swiss-Webster mice (1 mo old) were fed a basal vitamin E-deficient diet that contained either 8% fish oil + 2% corn oil or 10% lard with or without 1 g dl-alpha-tocopheryl acetate. The diets without vitamin E contained either 0.21 or 0.95 g ferric citrate/kg. Diets were fed for 4 wk/kg diet. Compared with the vitamin E-supplemented groups, mice fed diets without vitamin E (with or without supplemental iron) had significantly (P < 0.05) higher hepatic levels of thiobarbituric acid-reactive substances (TBARS), conjugated dienes and protein carbonyls when they were fed fish oil, but not lard. The levels of TBARS were further increased by iron supplementation in the mice fed fish oil. Significantly lower concentrations of alpha-tocopherol and higher glutathione (GSH) were found in the liver of mice fed fish oil and vitamin E than in those fed lard and vitamin E (P < 0.05). The activities of superoxide dismutase and glucose-6-phosphate dehydrogenase were lower in the fish oil-fed mice than in those fed lard (P < 0.05). The activities of Se-GSH peroxidase, non-Se-GSH peroxidase, catalase, and glutathione reductase were not altered by dietary fat or vitamin E/iron. The results obtained provide experimental evidence of the prooxidative effects of high dietary fish oil and iron, and suggest that vitamin E protects not only lipid-soluble compounds, but also water-soluble constituents, against oxidative damage. Further, dietary lipid plays a key role in determining cellular susceptibility to oxidative stress. PMID- 9202100 TI - Dietary linoleic acid intake controls the arterial blood plasma concentration and the rates of growth and linoleic acid uptake and metabolism in hepatoma 7288CTC in Buffalo rats. AB - In this study, we tested the hypothesis that dietary linoleic acid intake controls the arterial blood plasma linoleic acid concentration and the rates of tumor growth and linoleic acid metabolism in vivo. Seven groups of young male Buffalo rats (11-21 rats/group) were given free access to semipurified diets containing different amounts of corn and/or olive oils. Four other groups (7-11 rats/group) were 30% energy-restricted. Each experiment included periods for rat growth and plasma lipid stabilization (6 wk), measurement of mean daily arterial blood plasma fatty acid concentrations (3 wk), surgical implantation of a subcutaneous tissue-isolated hepatoma 7288CTC, tumor growth and harvest (2-4 wk). Linoleic + arachidonic acid (P = 0.007) and oleic acid (P = 0.002) concentrations in arterial blood plasma were increased as dietary intake of linoleic and oleic acids was increased, respectively. In rats given free access to food, tumor growth was directly dependent on the plasma concentrations of linoleic (P < 0.001) and arachidonic acids (P = 0.04). Tumor growth in energy-restricted rats was dependent only on the linoleic acid concentration (P = 0.008). Energy restriction itself caused a growth inhibition independent of plasma linoleic acid. The linoleic acid and total fatty acid concentrations of tumor triacylglycerols were directly dependent on the plasma linoleic acid concentration in rats given free access to food (P = 0.009). Hepatoma 7288CTC (both in vivo and during perfusion in situ) supported a dose-dependent conversion (P < 0.001) of plasma linoleic acid to the mitogen, 13-hydroxy-9, 11 octadecadienoic acid. We conclude that increased arterial blood plasma linoleic acid concentrations, caused by increased dietary intakes, specifically stimulate growth, lipid storage and linoleic acid metabolism in hepatoma 7288CTC in vivo. PMID- 9202101 TI - Caffeine enhances modulation of parasympathetic nerve activity in humans: quantification using power spectral analysis. AB - We investigated changes in autonomic nerve activity following caffeine intake by power spectral analysis of R-R intervals of heartbeats in humans. A beverage containing 240 mg of caffeine or a control beverage was given to 10 healthy volunteers, and R-R intervals were measured while subjects were sitting and controlling their respiration at a constant rate. After consumption of the caffeine-containing beverage, a transient and significant increase (P < 0.001) in spectral integrated values (areas under the curve) of high frequency power (high component, HC) was observed, and at 30 min the value was significantly greater than in controls (P < 0.02), suggesting an increase in vagal autonomic nerve activity. The effect of caffeine was also examined using decaffeinated coffee supplemented with exogenous caffeine (2 mg/kg body wt). A transient and significant increase (P < 0.0001) in HC was observed, and the value was significantly greater (P < 0.02) than when subjects consumed decaffeinated coffee without supplemental caffeine. The ratio of HC to total integrated value (which is also used as a selective indicator of vagal activity) was also significantly higher (P < 0.04) after caffeine consumption. Physiological variables accompanying the change in autonomic nerve activity (i.e., blood pressure, surface body temperature and heart rate) were not significantly affected by caffeine intake. These results suggest that power spectral analysis of heartbeat R-R intervals is an effective and noninvasive method for detecting subtle changes in autonomic nerve activity in response to food intake. PMID- 9202102 TI - Games people play with authors' names. PMID- 9202104 TI - US attacks EU gene-food labelling move. PMID- 9202103 TI - Congress and Varmus in clash over 'illegal' embryo research. PMID- 9202105 TI - NIH may drop special funds for 'new investigators'. PMID- 9202106 TI - Accord could lift block on European biotech patents. PMID- 9202107 TI - Japan reaches a compromise on organ transplants. PMID- 9202108 TI - India may set up genetics advisory panel. PMID- 9202109 TI - Blood lab boss jailed for neglect over HIV test. PMID- 9202110 TI - Cannibalism and kuru. PMID- 9202111 TI - You read it here first. PMID- 9202112 TI - Europe ambivalent on biotechnology. Biotechnology and the European Public Concerted Action group. PMID- 9202113 TI - Symmetry as destiny -- taking a balanced view of IQ. PMID- 9202114 TI - Potassium channels. Dendritic shock absorbers. PMID- 9202115 TI - Theoretical biology. A robust view of biochemical pathways. PMID- 9202116 TI - Cell biology. Explorers deliver tea to the pole. PMID- 9202117 TI - A peptide antibiotic from human skin. PMID- 9202118 TI - The zebrafish organizer requires chordino. PMID- 9202119 TI - K+ channel regulation of signal propagation in dendrites of hippocampal pyramidal neurons. AB - Pyramidal neurons receive tens of thousands of synaptic inputs on their dendrites. The dendrites dynamically alter the strengths of these synapses and coordinate them to produce an output in ways that are not well understood. Surprisingly, there turns out to be a very high density of transient A-type potassium ion channels in dendrites of hippocampal CA1 pyramidal neurons. These channels prevent initiation of an action potential in the dendrites, limit the back-propagation of action potentials into the dendrites, and reduce excitatory synaptic events. The channels act to prevent large, rapid dendritic depolarizations, thereby regulating orthograde and retrograde propagation of dendritic potentials. PMID- 9202120 TI - A new route for synthesis of dimethylsulphoniopropionate in marine algae. AB - The 3-dimethylsulphoniopropionate (DMSP) produced by marine algae is the main biogenic precursor of atmospheric dimethylsulphide (DMS). This biogenic DMS, formed by bacterial and algal degradation of DMSP, contributes about 1.5 x 10(13) g of sulphur to the atmosphere annually, and plays a major part in the global sulphur cycle, in cloud formation and potentially in climate regulation. Although DMSP biosynthesis has been partially elucidated in a higher plant, nothing is known about how algae make DMSP except that the whole molecule is derived from methionine. Here we use in vivo isotope labelling to demonstrate that DMSP synthesis in the green macroalga Enteromorpha intestinalis proceeds by a route entirely distinct from that in higher plants. From methionine, the steps are transamination, reduction and S-methylation to give the novel sulphonium compound 4-dimethylsulphonio-2-hydroxybutyrate (DMSHB), which is oxidatively decarboxylated to DMSP. The key intermediate DMSHB was also identified in three diverse phytoplankton species, indicating that the same pathway operates in other algal classes that are important sources of DMS. The fact that a transamination initiates this pathway could help explain how algal DMSP (and thereby DMS) production is enhanced by nitrogen deficiency. PMID- 9202121 TI - Corticofugal modulation of frequency processing in bat auditory system. AB - Auditory signals are transmitted from the inner ear through the brainstem to the higher auditory regions of the brain. Neurons throughout the auditory system are tuned to stimulus frequency, and in many auditory regions are arranged in topographical maps with respect to their preferred frequency. These properties are assumed to arise from the interactions of convergent and divergent projections ascending from lower to higher auditory areas; such a view, however, ignores the possible role of descending projections from cortical to subcortical regions. In the bat auditory system, such corticofugal connections modulate neuronal activity to improve the processing of echo-delay information, a specialized feature. Here we show that corticofugal projections are also involved in the most common type of auditory processing, frequency tuning. When cortical neurons tuned to a specific frequency are inactivated, the auditory responses of subcortical neurons tuned to the same frequency are reduced. Moreover, the responses of other subcortical neurons tuned to different frequencies are increased, and their preferred frequencies are shifted towards that of the inactivated cortical neurons. Thus the corticofugal system mediates a positive feedback which, in combination with widespread lateral inhibition, sharpens and adjusts the tuning of neurons at earlier stages in the auditory processing pathway. PMID- 9202122 TI - Congenital leptin deficiency is associated with severe early-onset obesity in humans. AB - The extreme obesity of the obese (ob/ob) mouse is attributable to mutations in the gene encoding leptin, an adipocyte-specific secreted protein which has profound effects on appetite and energy expenditure. We know of no equivalent evidence regarding leptin's role in the control of fat mass in humans. We have examined two severely obese children who are members of the same highly consanguineous pedigree. Their serum leptin levels were very low despite their markedly elevated fat mass and, in both, a homozygous frame-shift mutation involving the deletion of a single guanine nucleotide in codon 133 of the gene for leptin was found. The severe obesity found in these congenitally leptin deficient subjects provides the first genetic evidence that leptin is an important regulator of energy balance in humans. PMID- 9202123 TI - Fringe modulates Notch-ligand interactions. AB - The Notch family of transmembrane receptor proteins mediate developmental cell fate decisions, and mutations in mammalian Notch genes have been implicated in leukaemia, breast cancer, stroke and dementia. During wing development in Drosophila, the Notch receptor is activated along the border between dorsal and ventral cells, leading to the specification of specialized cells that express Wingless (Wg) and organize wing growth and patterning. Three genes, fringe (fng), Serrate (Ser) and Delta (Dl), are involved in the cellular interactions leading to Notch activation. Ser and Dl encode transmembrane ligands for Notch, whereas fng encodes a pioneer protein. We have investigated the relationship between these genes by a combination of expression and coexpression studies in the Drosophila wing. We found that Ser and Dl maintain each other's expression by a positive feedback loop. fng is expressed specifically by dorsal cells and functions to position and restrict this feedback loop to the developing dorsal ventral boundary. This is achieved by fng through a cell-autonomous mechanism that inhibits a cell's ability to respond to Serrate protein and potentiates its ability to respond to Delta protein. PMID- 9202124 TI - Robustness in simple biochemical networks. AB - Cells use complex networks of interacting molecular components to transfer and process information. These "computational devices of living cells" are responsible for many important cellular processes, including cell-cycle regulation and signal transduction. Here we address the issue of the sensitivity of the networks to variations in their biochemical parameters. We propose a mechanism for robust adaptation in simple signal transduction networks. We show that this mechanism applies in particular to bacterial chemotaxis. This is demonstrated within a quantitative model which explains, in a unified way, many aspects of chemotaxis, including proper responses to chemical gradients. The adaptation property is a consequence of the network's connectivity and does not require the 'fine-tuning' of parameters. We argue that the key properties of biochemical networks should be robust in order to ensure their proper functioning. PMID- 9202125 TI - A family of cytokine-inducible inhibitors of signalling. AB - Cytokines are secreted proteins that regulate important cellular responses such as proliferation and differentiation. Key events in cytokine signal transduction are well defined: cytokines induce receptor aggregation, leading to activation of members of the JAK family of cytoplasmic tyrosine kinases. In turn, members of the STAT family of transcription factors are phosphorylated, dimerize and increase the transcription of genes with STAT recognition sites in their promoters. Less is known of how cytokine signal transduction is switched off. We have cloned a complementary DNA encoding a protein SOCS-1, containing an SH2 domain, by its ability to inhibit the macrophage differentiation of M1 cells in response to interleukin-6. Expression of SOCS-1 inhibited both interleukin-6 induced receptor phosphorylation and STAT activation. We have also cloned two relatives of SOCS-1, named SOCS-2 and SOCS-3, which together with the previously described CIS form a new family of proteins. Transcription of all four SOCS genes is increased rapidly in response to interleukin-6, in vitro and in vivo, suggesting they may act in a classic negative feedback loop to regulate cytokine signal transduction. PMID- 9202126 TI - A new protein containing an SH2 domain that inhibits JAK kinases. AB - The proliferation and differentiation of cells of many lineages are regulated by secreted proteins known as cytokines. Cytokines exert their biological effect through binding to cell-surface receptors that are associated with one or more members of the JAK family of cytoplasmic tyrosine kinases. Cytokine-induced receptor dimerization leads to the activation of JAKs, rapid tyrosine phosphorylation of the cytoplasmic domains, and subsequent recruitment of various signalling proteins, including members of the STAT family of transcription factors, to the receptor complex. Using the yeast two-hybrid system, we have now isolated a new SH2-domain-containing protein, JAB, which is a JAK-binding protein that interacts with the Jak2 tyrosine-kinase JH1 domain. JAB is structurally related to CIS, a cytokine-inducible SH2 protein. Interaction of JAB with Jak1, Jak2 or Jak3 markedly reduces their tyrosine-kinase activity and suppresses the tyrosine-phosphorylation and activation of STATs. JAB and CIS appear to function as negative regulators in the JAK signalling pathway. PMID- 9202127 TI - Structure and function of a new STAT-induced STAT inhibitor. AB - The signalling pathway that comprises JAK kinases and STAT proteins (for signal transducer and activator of transcription) is important for relaying signals from various cytokines outside the cell to the inside. The feedback mechanism responsible for switching off the cytokine signal has not been elucidated. We now report the cloning and characterization of an inhibitor of STAT activation which we name SSI-1 (for STAT-induced STAT inhibitor-1). We found that SSI-1 messenger RNA was induced by the cytokines interleukins 4 and 6 (IL-4, IL-6), leukaemia inhibitory factor (LIF), and granulocyte colony-stimulating factor (G-CSF). Stimulation by IL-6 or LIF of murine myeloid leukaemia cells (M1 cells) induced SSI-1 mRNA expression which was blocked by transfection of a dominant-negative mutant of Stat3, indicating that the SSI-1 gene is a target of Stat3. Forced overexpression of SSI-1 complementary DNA interfered with IL-6- and LIF-mediated apoptosis and macrophage differentiation of M1 cells, as well as IL-6 induced tyrosine-phosphorylation of a receptor glycoprotein component, gp130, and of Stat3. When SSI-1 is overexpressed in COS7 cells, it can associate with the kinases Jak2 and Tyk2. These findings indicate that SSI-1 is responsible for negative-feedback regulation of the JAK-STAT pathway induced by cytokine stimulation. PMID- 9202128 TI - Repair of DNA loops involves DNA-mismatch and nucleotide-excision repair proteins. AB - A number of enzymes recognize and repair DNA lesions. The DNA-mismatch repair system corrects base-base mismatches and small loops, whereas the nucleotide excision repair system removes pyrimidine dimers and other helix-distorting lesions. DNA molecules with mismatches or loops can arise as a consequence of heteroduplex formation during meiotic recombination. In the yeast Saccharomyces cerevisiae, repair of mismatches results in gene conversion or restoration, and failure to repair the mismatch results in post-meiotic segregation (PMS). The ratio of gene-conversion to PMS events reflects the efficiency of DNA repair. By examining the PMS patterns in yeast strains heterozygous for a mutant allele with a 26-base-pair insertion, we find that the repair of 26-base loops involves Msh2 (a DNA-mismatch repair protein) and Rad1 (a protein required for nucleotide excision repair). PMID- 9202129 TI - Protein processing: a role in the pathophysiology of genetic disease. AB - Genetic diseases associated with an enzyme deficiency frequently have reduced intracellular levels of the mutant protein, despite apparently normal levels of message and protein synthesis. It has been suggested that the endoplasmic reticulum (ER) can recognise mutant protein as incorrectly folded and invoke 'quality control' processes which cause the retention and degradation of this protein. This process may occur, even for mutations which do not abrogate protein activity, contributing directly to pathophysiology. Genetic diseases associated with defects in ER and Golgi processing proteins have also been reported and generally result in impaired processing of multiple protein products. In this review the role of the ER and Golgi in the pathogenesis of genetic diseases relating to the vacuolar network are discussed. PMID- 9202130 TI - Analysis of the black-eyed pea trypsin and chymotrypsin inhibitor-alpha chymotrypsin complex. AB - The black-eyed pea trypsin and chymotrypsin inhibitor (BTCI) is a member of the Bowman-Birk protease inhibitor (BBI) family. The three-dimensional model of the BTCI-chymotrypsin complex was built based on the homology to Bowman-Birk inhibitors with known structures. An extensive theoretical and experimental study of these known structures has been performed. The model confirms the ideas about Bowman-Birk inhibitor structure-function relations and agrees well with our experimental data (circular dichroism, IR and light scattering). The electrostatic potentials at the enzyme-inhibitor contact surface reveal a pattern of complementary electrostatic potentials from which mutations can be inferred that could give these inhibitors an altered specificity. PMID- 9202131 TI - Asp-193 and Glu-218 of subunit II are involved in the Mn2+-binding of Paracoccus denitrificans cytochrome c oxidase. AB - Cytochrome c oxidase contains a binding site for a non-redox-active metal at the interface of subunits I and II, usually a magnesium ion. In Paracoccus denitrificans oxidase, typically 20% may be replaced by manganese, using standard growth media. Site-directed mutants were constructed in subunit II (D193N and E218Q), and the isolated enzymes analyzed by total-reflection X-ray fluorescence spectrometry and EPR. Both mutants show a strong reduction of the manganese stoichiometry and a diminished electron transfer activity, demonstrating that D193 and E218 are involved in the binding of a manganese/magnesium ion in this site. PMID- 9202132 TI - The endogenous cardiac sarcoplasmic reticulum Ca2+/calmodulin-dependent kinase is activated in response to beta-adrenergic stimulation and becomes Ca2+-independent in intact beating hearts. AB - We investigated the effects of beta-adrenergic stimulation on the activity of the endogenous cardiac sarcoplasmic reticulum Ca2+/calmodulin-dependent protein kinase (SRCaM kinase) in Langendorff-perfused rat hearts. We found that isoproterenol induced generation of autonomous (Ca2+-independent) SRCaM kinase activity to 28 +/- 4.4% of the total activity. Moreover, dephosphorylation of the autonomous SRCaM kinase with protein phosphatase 2A resulted in an enzyme that was again dependent on Ca2+ and calmodulin for its activity. Activation of SRCaM kinase was coupled to phospholamban phosphorylation and activation of the cAMP signaling system. Our results suggest that the cardiac SRCaM kinase is activated in response to beta-adrenoceptor stimulation. This activation stimulates autophosphorylation at its regulatory domain and converts it to an active Ca2+ independent species that may be the basis for potentiation of Ca2+ transients in the heart. PMID- 9202133 TI - Inhibition of the M1-->M2 (M(closed) --> M(open)) transition in the D96N mutant photocycle and its relation to the corresponding transition in wild-type bacteriorhodopsin. AB - Glutaraldehyde, lutetium ions and glycerol inhibit the blue shift of the difference spectra maximum of the M intermediate in the D96N mutant. The M formed has a spectrum indistinguishable from the M intermediate in wild-type bacteriorhodopsin. It has been concluded that the M(open) form previously described by us is identical to the M2 and Mn intermediates postulated by Zimanyi et al. (Photochem. Photobiol. (1992) 56, 1049-1055) and Sasaki et al. (J. Biol. Chem. (1992) 267, 20782-20786), respectively. It is supposed that its formation is accompanied by the appearance of the cytoplasmic proton half-channel. M(open) in the wild-type protein is present in a very low amount due to the shift of the M(closed) <--> M(open) equilibrium towards the M(closed). The inhibitors used do not prevent the multiphase pattern of the M formation in either mutant or wild type proteins. PMID- 9202134 TI - The vaccinia virus F17R protein interacts with actin. AB - We have examined the possible role of the F17R protein in vaccinia virus-induced rearrangements of the host actin cytoskeleton. F17R is localized to vaccinia induced actin tails late during infection. The recombinant vaccinia strain vRO11k is able to induce actin tails that are indistinguishable from controls in the absence of F17R expression. The association of vaccinia and myxoma virus F17R with the actin cytoskeleton in the absence of additional viral factors suggests a basic region in the N-terminal half of the protein is important for this interaction. A peptide corresponding to this region efficiently bundles actin filaments in vitro, confirming that the protein interacts directly with actin. Our results show F17R is not required for actin tail formation and highlight the difficulty in discriminating functional actin-binding proteins from those that associate by virtue of their basic nature. PMID- 9202136 TI - Single-strand-specific DNase activity is an inherent property of the 140-kDa protein of the snake venom exonuclease. AB - Polyclonal antibodies against the exonuclease from Crotalus adamanteus venom (the 140-kDa protein) inhibit both the exonucleolytic and the single-strand-specific endonucleolytic activities, present in the exonuclease preparation. The antibodies also diminish the ability of the enzyme to split the negatively supercoiled Bluescript KS+ in the AT-rich fragment near-by the transcription termination site of the Ampicillin gene. Therefore the single-strand-specific endonucleolytic activity was attributed to the protein molecule of the exonuclease. The processivity of the exonucleolytic action was found to be less than 3 monomers as indicated by the heparin trapping method. PMID- 9202135 TI - Endogenously produced peroxynitrite induces the oxidation of mitochondrial and nuclear proteins in immunostimulated macrophages. AB - Here we investigated the role of endogenous nitric oxide (NO) and peroxynitrite in the process of protein oxidation (as measured by the detection of 2,4 dinitrophenylhydrazine-reactive carbonyls) in immunostimulated macrophages. Immunostimulation of the macrophages by bacterial lipopolysaccharide and gamma interferon (LPS/IFNgamma) resulted in a marked increase in the oxidation of a large number of mitochondrial and nuclear proteins. The inhibitor of NO synthase, N(G)-methyl-L-arginine (3 mM), and the cell-permeable superoxide dismutase mimetic Mn(III)tetrakis(4-benzoic acid)porphyrin (300 microM) both reduced the extent of protein oxidation in response to LPS/IFNgamma. These results support the view that endogenously produced peroxynitrite induces protein oxidation in the mitochondria and nucleus of immunostimulated macrophages. PMID- 9202137 TI - Studies on a new series of THA analogues: effects of the aromatic residues that line the gorge of AChE. AB - A series of N-monoalkylsubstituted 1,2,3,4-tetrahydro-9-aminoacridines have been prepared after modelling simulation of the AChE-inhibitor complex. Molecular modelling has predicted a number of hydrophobic residues to be involved in the catalytic mechanism of this interaction between the binding sites of AChE and this series of aminoacridines. In these compounds the acridine moiety becomes sandwiched between the rings of PHE330 and TRP84. In particular, the alkyl chain shows the important role of aromatic groups as binding sites. Their in vitro inhibitory properties (enzyme from Electrophorus electricus) confirm the aromatic groups as a general and significant characteristic of the mechanism of AChE inhibition. PMID- 9202138 TI - Oxygen-dependent regulation of the respiration and growth of Escherichia coli by nitric oxide. AB - To elucidate the role of nitric oxide (NO) in the metabolisms of enteric bacteria, its effect on the respiration and growth of Escherichia coli was examined. Respiration of E. coli was reversibly inhibited by NO particularly under low oxygen tensions. Growth of E. coli was also inhibited by NO more strongly under low oxygen tension than at its high concentration. Because the intestinal lumen is anaerobic, even a small amount of NO might strongly inhibit the energy metabolism and growth of E. coli and other enteric bacteria in vivo than in air atmospheric conditions in which oxygen tension is unphysiologically high. PMID- 9202139 TI - Association of plant K+(in) channels is mediated by conserved C-termini and does not affect subunit assembly. AB - Inward rectifying potassium (K+(in)) channels play an important role in turgor regulation and ion uptake in higher plants. Here, we report a previously unrecognized feature of these proteins: K+(in) channel C-terminal polypeptides mediate channel protein interactions. Using a C-terminal fragment of potato guard cell K+(in) channel KST1 in a yeast two-hybrid screen two novel putative K+(in) channel proteins (SKT2 and SKT3) were identified by interaction of their C termini which contained a conserved domain (K(HA)). Interactions were confirmed by Western blot-related assays utilizing K+(in) channel C-termini fused to green fluorescence protein. Although deletion of the K(HA)-domain abolished these interactions, K+(in) currents were still detectable by patch-clamp measurements of insect cells expressing these KST1 mutants, indicating that formation of a functional channel does not depend on this C-terminal domain. PMID- 9202141 TI - Regulatory effects of aggregated LDL on apoptosis during foam cell formation of human peripheral blood monocytes. AB - In order to investigate the mechanisms how modified lipoproteins enhance foam cell formation, we cultured peripheral blood monocytes with various stimulants and examined the effects of aggregated low-density lipoprotein (agLDL) on cell viability and lipid metabolism. AgLDL could completely inhibit phorbol ester induced apoptosis, which was accompanied by intracellular cholesterol accumulation. Suppression of apoptosis-promoting proteases, ICE and CPP32, was observed in agLDL-treated cells. This indicates that agLDL accelerates foam cell formation through inhibition of apoptosis and enhancement of lipid accumulation in activated monocytes. By contrast, apoptosis was enhanced when monocytes were cultured with agLDL and M-CSF. Intracellular cholesterol accumulation was not significant in M-CSF treated cells. This suggests that M-CSF may act anti atherogenic through apoptotic elimination of lipid-baring macrophages and enhanced lipid turnover. Our observation supports the novel hypothesis that regulation of apoptosis may play an important role in the development of atherosclerosis. PMID- 9202140 TI - Insulin activates a PD 098059-sensitive kinase that is involved in the regulation of p70S6K and PHAS-I. AB - Incubating either Chinese hamster ovary (CHO) cells or 3T3-L1 adipocytes with insulin increased the phosphorylation of the eIF-4E-binding protein, PHAS-I. Insulin also activated p70S6K and the Erk-1 and Erk-2 isoforms of mitogen activated protein kinase (MAP kinase). However, the concentrations of the hormone needed to activate MAP kinase were 10-100 times higher than those needed to increase PHAS-I phosphorylation and p70S6K activity. Incubating cells with the inhibitor of MAP kinase kinase (MEK) activation, PD 098059, blocked the effects of low concentrations of insulin on PHAS-I and p70S6K. The effects of the inhibitor were overcome by increasing concentrations of insulin. The results indicate that insulin activates a PD 098059-sensitive kinase that is involved in the regulation of both p70S6K and PHAS-I. PMID- 9202142 TI - A natural motif approach to protein design: a synthetic leucine zipper peptide mimics the biological function of the platelet factor 4 protein. AB - The design of smaller functional mimics of large proteins has long been an important challenge. In this study we use the natural leucine zipper as a structural template to design a 31-residue peptide analog that mimics the function of the larger platelet factor 4 (PF4) protein. The heparin binding activity of PF4 has been introduced into an unrelated leucine zipper sequence only by virtue of incorporating four lysines of PF4. Circular dichroism and binding experiments have shown that the designed leucine zipper peptide adopts a stable helical conformation and shows significant PF4-like heparin binding activity. These results strongly suggest that the lysine residues play an important role in the binding of PF4 to heparin. The de novo generation of the PF4 function in a designed leucine zipper peptide demonstrates that the leucine zipper motif is a useful scaffold for the design of functional peptides and proteins. PMID- 9202143 TI - Structure of in-serum transfecting DNA-cationic lipid complexes. AB - Noticeable modifications of in-serum transfection efficiency of dioctadecylamidoglycyl-spermine (DOGS)-DNA complexes are observed, depending on DNA condensation conditions. The structures of the complexes are studied, keeping in mind the variability of lipid polymorphism, by cryo-transmission electron microscopy and X-ray diffraction. By increasing both pH and ionic strength, well organised lamellar structures with a period of 65 A replace supramicellar aggregates. A relationship between the structures and their in-vitro transfection activity is established. Efficiency in the presence of serum is maintained when a lamellar arrangement is involved. PMID- 9202144 TI - Phosphorylation of a 72-kDa protein in PDGF-stimulated cells which forms complex with c-Crk, c-Fyn and Eps15. AB - Ligand-induced activation of the beta-receptor for platelet-derived growth factor (PDGF) induces tyrosine phosphorylation of a number of downstream signaling proteins. In the present study, we used two-dimensional gel electrophoresis to characterize the spectrum of proteins phosphorylated in response to PDGF stimulation in porcine aortic endothelial cells expressing PDGF beta-receptors. Several previously known substrates for the PDGF beta-receptor were identified as well as a novel substrate of 72 kDa. The 72-kDa component could be co immunoprecipitated in complex with the adaptor protein c-Crk, the non-receptor tyrosine kinase c-Fyn and the signaling molecule Eps15. The results obtained suggests that the 72-kDa protein might play an important role in signaling via the PDGF beta-receptor, coupling non-receptor tyrosine kinases of the Src family with c-Crk and Eps15. PMID- 9202145 TI - The T transcription factor functions as a dimer and exhibits a common human polymorphism Gly-177-Asp in the conserved DNA-binding domain. AB - T is a transcription factor which activates transcription by binding to repeated arrangements of the dodecamer 5'-AGGTGTGAAATT-3'. Using in vitro synthesised T protein, we have demonstrated that T binds to its target DNA as a homodimer and that truncated protein containing only the N-terminal 233 amino-acid residues, which comprise the DNA-binding domain, can form a dimer. We also report a common human polymorphism, Gly-177-Asp, within the DNA-binding domain at a position which is a conserved glycine residue in T homologues from other vertebrates. The proposition that T forms heterodimers with other members of the T-box transcription factor family and the implications for disorders of axial development are discussed. PMID- 9202146 TI - Differential involvement of caspases in apoptosis of myeloid leukemic cells induced by chemotherapy versus growth factor withdrawal. AB - To assess the potential involvement of the caspase family in the IL-3-dependent murine myeloid leukemic cell line 32D, we studied the effect of bcl-2, crmA and three synthetic caspase inhibitors on apoptosis induced by chemotherapy or IL-3 withdrawal. Apoptosis induced by IL-3 deprivation or by ActD appears to be mediated by a crmA-insensitive pathway. Cell death by IL-3 withdrawal is inhibited by the caspase-inhibitor ZVAD-fmk, but not DEVD-fmk or YVAD-cmk. In contrast, DEVD-fmk as well as ZVAD-fmk protect 32D cells from ActD-induced apoptosis. These results indicate that different caspases are involved in apoptosis induced by growth factor withdrawal and by chemotherapy. PMID- 9202148 TI - Highly efficient control of iron-containing nitrile hydratases by stoichiometric amounts of nitric oxide and light. AB - The reaction of two iron-containing nitrile hydratases (NHase) with NO has been studied: NHase from Rhodococcus sp. R312, which is probably similar to the photosensitive N771 NHase, and the new NHase from Comamonas testosteroni NI1 whose aminoacid sequence is quite different from those of BR312 and N771 NHases. Both enzymes are equally inactivated after addition of stoichiometric amounts of NO added as an anaerobic solution or produced in situ under physiological conditions by a rat brain NO-synthase. Both enzymes are reactivated by photoirradiation, and two cycles of NO inactivation/photoactivation can be performed without significant loss of activity. Both iron-containing NHases have a high affinity for NO, similar to that of methemoglobin. PMID- 9202147 TI - Isolation of a cDNA coding for an ubiquitin-conjugating enzyme UBC1 of tomato- the first stress-induced UBC of higher plants. AB - A clone of an ubiquitin-conjugating enzyme (UBC) was isolated from a lambda-ZAP cDNA library, generated from mRNA of tomato (Lycopersicon esculentum) cells grown in suspension for 3 days. The open reading frame called LeUBC1, encodes for a polypeptide with a predicted molecular mass of 21.37 kDa, which was confirmed by bacterial overexpression and SDS-PAGE. Database searches with LeUBC1 showed highest sequence similarities to UBC1 of bovine and yeast. By Southern blot analysis LeUBC1 was identified as a member of a small E2 subfamily of tomato, presumably consisting of at least two members. As revealed by Northern blot analysis LeUBC1 is constitutively expressed in an exponentially growing tomato cell culture. In response to heat shock an increase in LeUBC1-mRNA was detectable. A strong accumulation of the LeUBC1-transcript was observed by exposure to heavy metal stress which was performed by treatment with cadmium chloride (CdCl2). The cellular uptake of cadmium was controlled via ICP-MS measurements. The data suggest that like in yeast, in plants a certain subfamily of UBC is specifically involved in the proteolytic degradation of abnormal proteins as result of stress. PMID- 9202149 TI - Mycothiol, 1-O-(2'-[N-acetyl-L-cysteinyl]amido-2'-deoxy-alpha-D-glucopyranosyl)-D myo-inositol, is the factor of NAD/factor-dependent formaldehyde dehydrogenase. AB - Two different NAD/coenzyme-dependent formaldehyde dehydrogenases exist, the well known NAD/GSH-dependent (EC 1.2.1.1) and the more recently discovered NAD/Factor dependent enzyme. The GSH-dependent one has been found in eukaryotes and Gram negative bacteria, the Factor-dependent one in two different Gram-positive bacteria. Previous work also showed that Factor and GSH are not interchangeable in the enzymatic reactions. Here it is revealed that the Factor is identical to mycothiol (MySH), 1-O-(2'-[N-acetyl-L-cysteinyl]-amido-2'-deoxy-alpha-D glucopyranosyl)-D- myo-inositol, a thiol compound which has recently been detected in Actinomycetes. Thus, MySH is GSH's companion as it is the coenzyme for the enzyme which henceforth can be indicated as NAD/MySH-dependent formaldehyde dehydrogenase. PMID- 9202150 TI - Tyrosine: an inhibitor of LDL oxidation and endothelial cell cytotoxicity initiated by superoxide/nitric oxide radicals. AB - Tyrosyl radicals can catalyze LDL oxidation. In addition to their LDL oxidizing ability, superoxide (O2.-)/nitric oxide (NO.) generate phenoxyl radicals when reacting with tyrosine. Therefore we tested if tyrosine can act as a pro-oxidant in O2.-/NO.-initiated LDL oxidation. When LDL was exposed to O2.-/NO., tyrosine exerted a strong inhibitory effect on O2.-/ NO.-initiated LDL oxidation as measured by TBARS formation and alteration in electrophoretic mobility of LDL. Tyrosine was also able to protect human endothelial cells from the cytotoxic effect of O2.-/NO.. Because O2.-/NO. can occur in vivo, the results may indicate that serum-free tyrosine could act as an efficacious physiological antioxidant in case of O2.-/NO.-initiated LDL oxidation and endothelial cell cytotoxicity. PMID- 9202151 TI - A hairpin-loop conformation in tandem repeat sequence of the ice nucleation protein revealed by NMR spectroscopy. AB - The 1H-NMR spectrum of a synthetic 24-residue peptide (A1-G-V-D-S-S-L-I-A-G-Y-G-S T-Q-T-S-G-S-D-S-A-L-T24; INP24), comprising three repeats of the 8-residue consensus sequence of Pseudomonas syringae ice nucleation protein, was fully assigned using 2-dimensional (2D) NMR spectroscopy at 4 degrees C and 30 degrees C. Close proximity of the aliphatic protons between Leu7, Ile8, Ala9, and the ring-protons of Tyr11 was indicated from the observation of the inter-molecular nuclear Overhauser enhancement (NOE) effect. Hydrogen-bonding was strongly suggested for the NH group of Leu7 from its extremely low-temperature coefficient estimated from the temperature dependence of the chemical shift. These results indicate the formation of a hairpin-loop conformation constructed by a hexapeptide segment of INP24, -Leu7-Ile8-Ala9-Gly10-Tyr11-Gly12. PMID- 9202152 TI - Subunit stoichiometry of the pancreatic beta-cell ATP-sensitive K+ channel. AB - We have investigated the subunit stoichiometry of the pancreatic beta-cell ATP sensitive K+ (KATP) channel (SUR1/Kir6.2 channel) by constructing cDNA encoding a single polypeptide (beta alpha polypeptide) consisting of a SUR1 (beta) subunit and a Kir6.2 (alpha) subunit. 86Rb+ efflux and single-channel properties of COS1 cells expressing beta alpha polypeptides were similar to those of COS1 cells coexpressing alpha monomers and beta monomers. Coexpression of beta alpha polypeptides with alpha monomers inhibited the K+ currents, while coexpression with beta monomers did not. We then constructed another single polypeptide (beta alpha2) consisting of a beta subunit and a dimeric repeat of the alpha subunit. 86Rb+ efflux from COS1 cells expressing beta alpha2 polypeptides was small, but was restored by supplementation with beta monomers. These results indicate that the activity of K(ATP) channels is optimized when the alpha and beta subunits are coexpressed with a molar ratio of 1:1. Since inward rectifier K+ channels are thought to function as homo- or hetero-tetramers, this suggests that the K(ATP) channel functions as a multimeric protein, most likely a hetero-octamer composed of a tetramer of the Kir6.2 subunit and a tetramer of the SUR1 subunit. PMID- 9202153 TI - Identification of a novel nuclear localization signal in Sam68. AB - Sam68, a nuclear RNA binding protein, binds to Src and is phosphorylated at tyrosine residues in an M-phase specific manner. Here we identified a stretch of 24 amino acid residues in the COOH-terminal portion of Sam68 which function as a nuclear localization signal. This signal sequence bears no apparent homology to any other known nuclear localization sequence. However, this sequence was found to contain a motif, PPXXR (P, Pro; R, Arg), which is conserved in various RNA binding proteins including hnRNP proteins. Replacement of Arg in this motif with Ala abolished the nuclear accumulation of a GFP fusion protein, suggesting that this residue is important in translocating the protein to the nucleus. PMID- 9202154 TI - Inverse gene expression of prostacyclin and thromboxane synthases in resident and activated peritoneal macrophages. AB - Prostacyclin and thromboxane A2 produced from prostaglandin H2 are known to be important modulators with opposite biological activities. To examine possible roles of these prostanoids in immune responses, we have studied the gene expression of prostacyclin synthase (PGIS) and thromboxane synthase (TXS) in murine resident macrophages or in macrophages elicited with casein or bacillus Calmette-Guerin (BCG). Northern blot analyses showed that the PGIS mRNA was expressed in a decreasing order in the resident, and casein- and BCG-elicited macrophages. In contrast, the TXS mRNA was expressed in an increasing order in the resident, and casein- and BCG-elicited macrophages. On the other hand, the mRNA for cyclooxygenase-2, which produces PGH2 and participates in the production of prostanoids in inflammation, was expressed in both the resident and BCG elicited macrophages but barely in the casein-elicited cells. In situ hybridization analysis showed that the expression of mRNAs for PGIS and TXS was ascribable not only to the alteration of the expression levels of both mRNAs in the each macrophage but also to the changes in subpopulations of the cells expressing these mRNAs. These observations suggested that the inverse gene expression of PGIS and TXS in macrophages contributes to immune responses by modulating the relative levels of prostacyclin and thromboxane A2. PMID- 9202155 TI - The photoreceptor rim protein is an ABC transporter encoded by the gene for recessive Stargardt's disease (ABCR). AB - Rim protein (RmP) is a high-Mr membrane glycoprotein that has been localized to the rims of photoreceptor outer segment discs, but its molecular identity is unknown. Here, we describe the purification of RmP and present the sequence of its mRNA. RmP is a new member of the ATP-binding cassette (ABC) transporter superfamily. We show that RmP is expressed specifically in photoreceptors and predominantly in outer segments. Further, RmP is identical to the protein recently shown to be affected in recessive Stargardt's disease. RmP is the first ABC transporter observed in photoreceptors and may play a role in the photoresponse. PMID- 9202156 TI - cAMP stimulates protein kinase B in a Wortmannin-insensitive manner. AB - Activation of protein kinase B (PKB) by growth factors has been demonstrated to proceed via phosphatidylinositol 3-kinase (PI3-kinase). Here, we show that agents which raise intracellular cAMP can also stimulate PKB. However, this effect is not sensitive to wortmannin, indicating that it is PI3-kinase independent. This activation does not appear to result from direct phosphorylation by protein kinase A (PKA) since GST-PKB is not an effective PKA substrate. In addition, the activation pathway of PKB by cAMP seems to be linked to that of growth factors, albeit downstream of PI3-kinase. Evidence for this is that a constitutive active PKB, T308D, S473D, containing activating mutations in the serine and threonine residues which are phosphorylated subsequent to PI3-kinase activation, cannot be further stimulated by cAMP elevations. Hence, these data suggest that, in addition to growth factors, cAMP can also lead to activation of PKB. This cAMP stimulatory action appears to require phosphorylation of T308 and S473, and hence would indicate that cAMP modulates the phosphorylation event of these PKB regulatory sites. PMID- 9202157 TI - Molecular cloning of a diacylglycerol kinase isozyme predominantly expressed in rat retina. AB - We have cloned and characterized a new diacylglycerol kinase (DGK) isozyme which is expressed in the retina and the brain of rat. The cDNA contains an open reading frame of 567 amino acid residues with a predicted protein of 64 kDa and shows very high homology to human DGK epsilon. The new DGK isozyme contains two distinctive zinc-finger structures and a putative catalytic domain. This DGK expressed predominantly in the inner and outer nuclear layers of retina. This expression pattern is different from those of the previously cloned DGKs including the human DGK epsilon, suggesting that this DGK isozyme has potential importance in visual functions as was the case in Drosophila retinal cells. PMID- 9202159 TI - Bacterial expression and purification of biologically active mouse c-Fos proteins by selective codon optimization. AB - A simple strategy using selective codon optimization was devised to express mouse c-Fos protein in high levels in E. coli. Ten arginine codons located in the basic region were optimized to achieve high levels of protein expression. The c-Fos protein was purified to near homogeneity and was demonstrated to be biologically active by assaying its several biological activities. PMID- 9202158 TI - Antioxidant activity of allopurinol on copper-catalysed human lipoprotein oxidation. AB - We found that allopurinol, at therapeutically relevant concentrations (9-58 microM), significantly counteracted copper-catalysed human non-HDL lipoprotein oxidation, as assessed by thiobarbituric acid reactant content and kinetics of conjugated diene formation. Oxypurinol was ineffectual. Both drugs had no activity on metal-independent, peroxyl radical-induced lipoprotein oxidation. Specific fluorescence-quenching experiments revealed that only allopurinol could interact with copper antagonizing metal binding to lipoproteins. Thus, therapeutic allopurinol concentrations can inhibit copper-catalysed lipoprotein oxidation through metal complexation, suggesting some antioxidant-antiatherogenic activity of the drug in vivo. PMID- 9202160 TI - Involvement of soluble proteinous factors in auxin-induced modulation of P-type ATPase in rice (Oryza sativa L.) seedlings. AB - The marked difference in auxin sensitivity between plant roots and shoots was studied in terms of auxin-induced stimulation of membrane P-type ATPase. The results suggest the existence in rice seedlings of, at least, two isoforms of soluble proteinous factors (SPF), SPF(I) and SPF(II), which are involved in the stimulatory action of auxin on the enzyme. It also is indicated that SPF(I) which mediates the auxin effect in low hormone concentration range (10(-10)-10(-7) M IAA) is the dominant isoform in roots, whereas SPF(II) which does in high hormone concentration range (10(-7)-10(-4) M IAA) is that in shoots. PMID- 9202161 TI - Molecular cloning and expression of a bovine endothelial inward rectifier potassium channel. AB - A 5.1 kb cDNA encoding an inward rectifier K+ channel (BIK) was isolated from a bovine aortic endothelial cell library. The cDNA codes for a 427-amino-acid protein with two putative transmembrane regions. Sequence analysis reveals that BIK is a member of the Kir2.1 family of inward rectifier K+ channels. Expression in Xenopus oocytes showed that BIK is a K+-specific strong inward rectifier channel that is sensitive to extracellular Ba2+, Cs+, and a variety of anti arrhythmic agents. Northern analysis revealed that endothelial cells express a 5.5 kb BIK mRNA that is sensitive to shear stress. PMID- 9202162 TI - Oligomer formation of histamine H2 receptors expressed in Sf9 and COS7 cells. AB - A histamine H2 receptor, which had been mutated at its glycosylation site and tagged at its N-terminus with an HA tag (HA-H2 receptor), was expressed in Sf9 cells and COS7 cells. Immunoprecipitation and immunoblotting of HA-H2 receptors with alphaHA antibody revealed four bands of 31.5 +/- 2.5 kDa, 59.0 +/- 6.0 kDa, 80.5 +/- 4.5 kDa and 120 kDa. These bands were also detected by immunoblot using anti-H2 receptor serum (C-terminus). In addition, H2 receptors without the HA-tag coimmunoprecipitated with HA-tagged H2 receptors devoid of the 51 C-terminal amino acids, via immunoprecipitation with alphaHA antibody, when the two receptors were coexpressed. These results suggest that H2 receptors are present as receptor oligomers and that the C-terminal portion is not involved in the formation of these oligomers. PMID- 9202164 TI - An antagonist of ATP-regulated potassium channels, the guanidine derivative U 37883A, stimulates the synthesis of phosphatidylserine in rat liver endoplasmic reticulum membranes. AB - The guanidine derivative U-37883A has been found to stimulate in vitro synthesis of phosphatidylserine in endoplasmic reticulum membranes, catalyzed exclusively by a serine-specific base exchange enzyme. The stimulation of the enzyme activity by the drug was concentration-dependent, with EC50 of 54 microM, while the biologically inactive analog of U-37883A, U-42069, was without effect. The stimulation caused by U-37883A was enhanced under the conditions when active transport of Ca2+ into the lumen of microsomal vesicles was induced, whereas it was inhibited by a calcium ionophore, A23187, and by a specific inhibitor of Ca2+ ATPase, thapsigargin. On the other hand, a potassium ionophore, valinomycin, had no effect on phosphatidylserine synthesis. U-37883A did not affect the Km of the base exchange enzyme for serine, but greatly reduced the EC50 value of the enzyme for calcium. Furthermore, Ca2+ uptake by endoplasmic reticulum vesicles has been found to increase in the presence of U-37883A. These observations suggest that U 37883A enhances phosphatidylserine synthesis indirectly by acting on calcium transport, thus affecting calcium concentration within the lumen of endoplasmic reticulum membranes. Alternatively, the effect of the drug could be propagated via the mechanism by which phospholipid flip-flop movement, known to regulate the serine-specific base exchange reaction, is modulated. PMID- 9202163 TI - Biomarker evidence of DNA oxidation in lung cancer patients: association of urinary 8-hydroxy-2'-deoxyguanosine excretion with radiotherapy, chemotherapy, and response to treatment. AB - Ratios of urinary 8-hydroxy-2'-deoxyguanosine to urinary creatinine (8 OHdG/creatinine) have been considered as a good biological indicator of DNA oxidation. Urinary 8-OHdG/creatinine levels of lung cancer patients were evaluated by enzyme-linked immunosorbent assay using a monoclonal antibody N45.1 during radiotherapy and chemotherapy. An increase in urinary 8-OHdG/creatinine was found in non-small-cell carcinoma (non-SCC) patients during the course of radiotherapy. SCC patients showed higher levels of urinary 8-OHdG/creatinine than the controls. Furthermore, SCC patients with complete or partial response to the chemotherapy showed a significant decrease in urinary 8-OHdG/creatinine while patients with no change or progressive disease showed an increase. PMID- 9202165 TI - Non-iron porphyrins inhibit beta-haematin (malaria pigment) polymerisation. AB - Infrared spectroscopy was used to evaluate the effect of non-iron porphyrins (protoporphyrin IX and haematoporphyrin) on haematin polymerisation to beta haematin at acidic pH. Both molecules effectively inhibited the reaction, with haematoporphyrin 6 times as active as protoporphyrin IX. We postulated that the interaction between the pi electron system of porphyrin rings leads to the formation of pi-pi adducts, which inhibit polymer elongation in the same way as antimalarial drugs (e.g., chloroquine); the presence of hydroxyl groups able to bind haem iron enhances activity. PMID- 9202166 TI - Interaction of annexins IV and VI with ATP. An alternative mechanism by which a cellular function of these calcium- and membrane-binding proteins is regulated. AB - Annexin VI from porcine liver can be photoaffinity-labeled with 8-azido-[gamma 32P]ATP in a concentration-dependent, saturable manner. The extent of labeling varied with the concentration of calcium. The dissociation constant for the nucleotide was found to be in the range reported for ATP-binding proteins. The ATP analog, 2'-(or 3')-O-(2,4,6-trinitrophenyl)-adenosine 5'-triphosphate, also bound to AnxVI, as indicated by shift in its fluorescence spectra in the presence of protein. Any significant 8-azido-ATP or TNP-ATP binding was not observed with AnxIV. ATP modulated the binding of AnxVI to erythrocyte membrane and increased the Ca2+ concentration required for half-maximal binding of AnxVI to F-actin. PMID- 9202167 TI - Modulation of intracellular protein degradation by SSB1-SIS1 chaperon system in yeast S. cerevisiae. AB - In prokaryotes, DnaK-DnaJ chaperon is involved in the protein degradation catalyzed by proteases La and ClpA/B complex as shown in E. coli. To extend this into eukaryotic cells, we examined the effects of hsp70 genes, SSA1 and SSB1, and DnaJ genes, SIS1 and YDJ1, on the growth of proteasome subunit mutants of the yeast S. cerevisiae. The results identified SSB1 and SIS1 as a pair of chaperon genes specifically involved in efficient protein turnover in the yeast, whose overexpression suppressed the growth defects caused by the proteasome mutations. Moreover, a single amino acid substitution in the putative peptide-binding site of SSB1 protein profoundly enhanced the suppression activity, indicating that the activity is mediated by the peptide-binding activity of this chaperon. Thus SSB1, with its partner DnaJ, SIS1, modulates the efficiency of protein turnover through its chaperon activity. PMID- 9202168 TI - The nodulation gene nolK of Azorhizobium caulinodans is involved in the formation of GDP-fucose from GDP-mannose. AB - The nolK gene of Azorhizobium caulinodans is essential for the incorporation of a fucosyl group in Nod factors. A NAD(P)-binding site is present in the NolK amino acid sequence and the gene is homologous to Escherichia coli genes, presumably involved in GDP-fucose synthesis. Protein extracts of A. caulinodans, overexpressing nolK, have an enzyme activity that synthesizes GDP-fucose from GDP mannose. nolK most probably encodes a 4-reductase performing the last step in GDP fucose synthesis. Wild-type A. caulinodans produces a population of fucosylated and non-fucosylated molecules but the nolK-overexpressing strain produces only fucosylated Nod factors. Thus, the production of activated fucosyl donors is a rate-limiting step in Nod factor fucosylation. PMID- 9202169 TI - Cloning, sequencing and expression in MEL cells of a cDNA encoding the mouse ribosomal protein S5. AB - We describe the isolation and characterization of a cDNA encoding the mouse S5 ribosomal protein. It was isolated from a MEL (murine erythroleukemia) cell cDNA library by differential hybridization as a down regulated sequence during HMBA induced differentiation. Northern series analysis showed that S5 mRNA expression is reduced 5-fold throughout the differentiation process. The mouse S5 mRNA is 760 bp long and encodes for a 204 amino acid protein with 94% homology with the human and rat S5. PMID- 9202170 TI - Actin assembly by cadmium ions. AB - Cadmium is a highly toxic metal entering cells by a variety of mechanisms. Its toxic action is far from being completely understood, although specific interaction with the cellular calcium metabolism has been indicated. Metal ions that influence intracellular Ca2+ concentrations or compete with Ca2+ for protein binding sites may exert an effect on actin filaments, whose assembly and disassembly are both regulated by a number of calcium-dependent factors. Cadmium is such a metal. Much evidence demonstrates that cadmium interferes with the dynamics of actin filaments in various types of cells. Here we show that, at high (0.8-1.0 mM) concentrations, CdCl2 causes actin denaturation. At such Cd2+ concentrations, actin precipitates (really actin, as shown by SDS-PAGE, see Fig. 1B) in the form of irregular, disordered clots, clearly appreciable by electron microscopy. Denaturation seems to be reversible since, after Cd2+ removal by dialysis, the polymerizability of sedimented actin is restored almost completely. On the other hand, at concentrations ranging from 0.25 to 0.6 mM, CdCl2 is more effective as an actin polymerizing agent than both MgCl2 and CaCl2. The Cd related increase in the actin assembly rate is ascribable to an enhanced nucleation rather than to an increased monomer addition to filament growing ends. The latter, in contrast, appears quite slow. Critical concentration measurements revealed that the extent of polymerization of both Mg- and Cd-assembled actin are very close (C(c) ranges from 0.25 to 0.5 microM), while Ca-polymerized actin shows a polymerization extent markedly lower (C(c) = 4.0 microM). By both the fluorescent Ca2+ chelator Quin-2 assay and limited proteolysis of actin by trypsin and alpha-chymotrypsin, the real substitution of G-actin-bound Ca2+ by Cd2+ has been appreciated. The increase in Quin-2 fluorescence after addition of excess CdCl2 indicates that, in our experimental conditions, Ca2+ tightly-bound to actin is partially (60-70%) replaced by Cd2+, forming Cd-actin. Electrophoretic patterns after limited proteolysis reveal that the trypsin cleavage sites in the segment 61-69 of the actin polypeptide chain are less accessible in Cd-actin than in Ca-actin, although the cation-dependent effect is less pronounced in Cd-actin than in Mg-actin. Our results are consistent with some of the consequences on microfilament organization observed in Cd2(+)-treated cells; however, considering the positive effect of Cd2+ on actin polymerization in solution we have noticed that this was never observed in vivo. A different indirect effect of Cd2+ on some cellular event(s) influencing cytoplasmic actin polymerization appears to be reasonable. PMID- 9202171 TI - Renal epidermal growth factor precursor: proteolytic processing in an in vitro cell-free system. AB - The enzymatic processing of the membrane-bound renal epidermal growth factor precursor (proEGF) could be an important step in the control of nephrogenic repair consecutive to kidney insult. The enzyme machinery responsible for that processing was examined in a cell-free system consisting of renal membranes isolated from kidney homogenates by differential centrifugation, and incubated in vitro. After a 24-h incubation at 37 degrees C, 6-14% of membrane-bound proEGF was processed and soluble products with EGF immunoreactivity were released. As revealed by HPLC and Western blotting analysis, the products of proEGF proteolysis consisted of 6 kDa EGF (the molecular weight of mature EGF) and two polypeptides with molecular weights around 45 kDa. Interestingly the 45 kDa EGF forms, like the 6 kDa EGF, exhibited mitogenic activity toward growth-arrested NRK-52E renal cell line. The kinetic study of proEGF degradation gave data consistent with the 45 kDa product(s) being processing intermediate(s) between proEGF and 6 kDa EGF. The enzymatic activity responsible for proEGF nicking was inhibited by divalent heavy metal ions (Cu2+ or Zn2+) and several protease inhibitors (aprotinin, PMSF, leupeptin, soybean trypsin inhibitor), suggesting that proEGF is processed by kallikrein-like serine proteases present in the membrane preparations. Along with previous studies, the current observations suggest that renal kallikreins might play a role in renal tubular regeneration by promoting the release of soluble EGF in renal tissue. PMID- 9202172 TI - Induction of the stress protein Grp75 by amino acid deprivation in CHO cells does not involve an increase in Grp75 mRNA levels. AB - The induction of the stress protein Grp75 in response to amino acid deprivation of Chinese Hamster Ovary cells was characterised using a specific monoclonal antibody. A 2-fold increase in the Grp75 protein content occurred over a period of 5-10 h after incubation of the cells in amino acid-free medium. A partial induction was obtained when either all non-essential amino acids or all essential amino acids were omitted from the medium indicating a broad-specificity response. Deletion of the single amino acids tryptophan, histidine or phenylalanine from otherwise complete medium also produced a partial induction of the protein. The increase in the level of Grp75 was completely blocked by cycloheximide, but only partially blocked by the inhibitors of mRNA synthesis actinomycin D and alpha amanitin. A specific cDNA probe for Grp75 was generated by PCR and used to quantify mRNA levels. No increase in Grp75 mRNA was observed during the induction of the protein indicating that the primary regulation of Grp75 expression was not at the transcriptional level. These results contrast with the large increase in asparagine synthetase mRNA which has been shown to occur during amino acid deprivation, and indicate that cells respond to this form of stress by more than one mechanism. PMID- 9202173 TI - Osmo-stress-induced changes in neutral trehalase activity of the fission yeast Schizosaccharomyces pombe. AB - Exposure of repressed growing cultures of Schizosaccharomyces pombe to various extracellular concentrations of NaCl, sorbitol or glycerol resulted in a reversible increase in neutral trehalase activity which was maintained while the cells were in the presence of high environmental osmolarity. Treatment of osmo stress-induced trehalase by phosphatase lead to a decreased activity indicating that the active enzyme is phosphorylated. The stress response following the osmotic shock required protein synthesis and was independent of the cAMP dependent protein kinase pathway. Cells disrupted for wis] or phh1 (identical to sty1 and spc1), which encode members of the mitogen-activated protein kinase (MAPK) cascade, showed that the osmo-stress-induced increase in trehalase markedly diminished. In contrast, the heat shock-induced increase in trehalase remained unchanged in these cells. Taken together, the data suggest that the elevation of trehalase activity in Schiz. pombe under conditions of high osmolarity is due to de novo synthesis of the enzyme and that this process is modulated through a MAPK signal transduction pathway as part of the physiological response to the osmotic stress. The wisl-phhl MAPK cascade, however, does not appear to form part of the mechanism underlaying the increase in trehalase after heat stress. PMID- 9202174 TI - Dichloroaromatic phosphoguanidines are potent inhibitors but very poor substrates for cytosolic creatine kinase. AB - New phosphorylated guanidines have been synthesized and examined as potential inhibitors for creatine kinase. These compounds show a significant increase of inhibitory activity in comparison with the corresponding guanidines. Unlike the guanidines, they are competitive inhibitors because of the phosphoryl group. N Phospho-N'-2-(2,6-dichlorophenyl)ethylguanidine is a potent inhibitor (K(i) = 2.0 mM and 1.2 mM respectively for muscle and brain-type creatine kinase). Although these phosphorylated analogs of creatine phosphate have a very poor substrate activity in the reverse reaction, the phosphoryl group is important for binding to the enzyme. PMID- 9202175 TI - Production and characterization of a mutant cell line defective in aminophospholipid translocase. AB - Phospholipids are normally asymmetrically distributed between leaflets of the plasma membrane, due to the activity of aminophospholipid translocase (APT), a putative plasma membrane Mg2(+)-ATPase which is thought to selectively transport phosphatidylserine (PS) and other aminophospholipids from outer to inner membrane leaflet. Although several candidate proteins have been proposed to serve this function, positive identification awaits demonstration of their capacity to restore APT activity to a cell line that is deficient in this process. This study describes a simple and rapid protocol for the production and selection of mutant cell lines that are defective in APT activity and suitable for expression cloning of cDNAs coding for candidate APT enzymes. By flow cytometry, we demonstrate the time-dependent uptake of NBD-labeled PS, but not phosphatidylcholine (PC), by the mouse fibroblast cell line SV-T2. This uptake was inhibited by known inhibitors of APT, including o-vanadate and N-ethylmaleimide, and by ATP-depletion. SV-T2 cells were mutagenized with ethyl methanesulfonate, and APT-deficient cells were isolated by fluorescence activated cell sorting using NBD-PS as substrate. From a total of 7.2 x 10(6) cells passed through the flow cytometer, 98 clones exhibited APT activity that was less than 50% of that observed for wild-type SV-T2 cells. One clone which exhibited < or = 25% of that observed for wild-type cells, mutant M2711, was further characterized. The defect in mutant M2711 was specific for NBD PS, and cellular ATP was unchanged, suggesting that the defect in APT activity was not due to a decrease in cellular ATP levels. Mutant M2711 exhibited a growth pattern indistinguishable from that of wild-type SV-T2 cells, and SV-40 large T antigen, which is needed for efficient episomal replication of plasmids containing the SV40 origin of replication, was unchanged. Finally, transfection of M2711 with cDNAs for marker membrane proteins consistently resulted in the same high level of protein expression as that observed for identically transfected wild-type SV-T2. Thus, flow cytometry can be used for rapid identification of mutants with defects in phospholipid transport that are suitable for functional reconstitution by transfection with candidate APT cDNAs. PMID- 9202176 TI - Angiotensin II receptors, AT1 and AT2 in the rat epididymis. Immunocytochemical and electrophysiological studies. AB - Previous work from our laboratory has provided evidence for the presence of a tissue renin-angiotensin system in the rat epididymis. In the current investigation, the regional localization of angiotensin II receptors, type I (AT1) and type II (AT2) was studied immunocytochemically using specific anti peptide antibodies against the second extracellular loops of AT1 and AT2 receptors, and pharmacologically using specific receptor antagonists in conjunction with the short-circuit current technique. The immunocytochemical results showed that AT1 and AT2 immunoreactivities were predominantly localized in the basal region of the epididymal epithelium. Electrophysiological studies using the short-circuit current technique demonstrated a stimulatory effect of basolaterally applied angiotensin II on the epididymal electrogenic ion transport. This effect was inhibitable by the addition of AT1 antagonist, losartan but not by AT2 antagonist, PD123177, indicating a functional role of AT1 in epididymal electrolyte transport. The present finding suggests that angiotensin II receptors may play an important role in the regulation of epididymal function. PMID- 9202177 TI - Regulation of calcium-induced exocytosis from gastric chief cells by protein phosphatase-2B (calcineurin). AB - The molecular mechanisms whereby calcium stimulates secretion are uncertain. In the present study, we used streptolysin O (SLO)-permeabilized chief cells from guinea pig stomach to investigate whether protein phosphatase-2B (calcineurin), a calcium/calmodulin-dependent, serine/threonine phosphatase plays a role in mediating calcium-induced pepsinogen secretion. Preincubation of cells with alpha naphthylphosphate, a non-specific phosphatase inhibitor, decreased calcium induced secretion. Likewise, specific inhibitors of protein phosphatase-2B (cyclosporin-A and FK-506) caused a dose-dependent reduction in calcium-induced pepsinogen secretion. Moreover, in intact cells, cyclosporin-A and FK-506 inhibited pepsinogen secretion caused by cholecystokinin, carbamylcholine and A23187, agonists known to increase chief cell cytosolic calcium. Okadaic acid, an inhibitor of protein phosphatase-1 and -2A, had no effect on secretion caused by these agonists. Chief cell calcium-dependent phosphatase activity, measured using radiolabeled casein as substrate, was reduced selectively by inhibitors of protein phosphatase-2B. Endogenous substrates for calcium/calmodulin-dependent phosphatase activity were identified by analyzing chief cell lysates using 2 dimensional gel electrophoresis. Increasing the cytosolic calcium concentration resulted in dephosphorylation of a 55-kDa, acidic cytoskeletal protein. FK-506 inhibited dephosphorylation of this protein. Thus, in permeabilized chief cells, specific inhibitors of protein phosphatase-2B inhibit calcium-induced pepsinogen secretion, calcium/calmodulin-dependent phosphatase activity and calcium-induced dephosphorylation of a 55-kDa, acidic cytoskeletal protein. These results support the hypothesis that protein phosphatase-2B (calcineurin) plays an important role in mediating calcium-induced exocytosis. PMID- 9202178 TI - Binding of two fluorescent cAMP analogues to type I and II regulatory subunits of cAMP-dependent protein kinases. AB - Binding of two long wavelength fluorescent cAMP analogues, 8-thioacetamido fluorescein-cAMP (SAF-cAMP) and 8-thioacetamido-rhodamine-cAMP (SAR-cAMP), to the RI (from bovine muscle) and RII (from bovine heart) regulatory subunits of cAMP dependent kinases has been studied. Displacement of [3H]cAMP from RI and RII and equilibrium dialysis measurements show that the fluorescent nucleotides are high affinity ligands for the cAMP binding sites. The binding is characterized by complex fluorescence spectral and fluorescence anisotropy changes, more evident for the fluorescein than for the rhodamine derivative. The fluorescence excitation spectrum of the bound SAF-cAMP is characterized by the appearance of a red shifted shoulder at 500-510 nm excitation wavelength region. Any change of the bound/free ratio in a solution equilibrium is accompanied by changes in fluorescence and anisotropy signals which are best detected at suitable wavelengths. It is proposed that fluorescence and anisotropy changes can distinguish between binding to type B (slow dissociating) and A (fast dissociating) cAMP binding sites of regulatory subunits. Applications of the fluorescent nucleotides to kinase localization and cAMP determination in living cells are discussed. PMID- 9202179 TI - Induction of embryonal carcinoma cell differentiation by deferoxamine, a potent therapeutic iron chelator. AB - We investigated the effects of deferoxamine on the differentiation of embryonal carcinoma F9 cells. Deferoxamine, a widely used therapeutic agent for thalassemia and iron overload, was found to induce F9 cell differentiation and to have some unique characteristics compared with other chelators, hinokitiol and dithizone, which were previously reported to induce differentiation of these cells. This hydrophilic agent induced reversible differentiation as did sodium butyrate, whereas other chelators did not. However, morphological features of the cells after deferoxamine-induced differentiation were similar to those of cells incubated with the other chelators. The differentiation-inducing activity of deferoxamine was abolished by preincubation with Fe3+ ions, similarly to the other chelators examined. Moreover, cell proliferation was inhibited by treatment with this agent, and the numbers of cells in the colonies were reduced by apoptosis. Based on these results, we conclude that deferoxamine induces differentiation and apoptosis of F9 cells via chelation of extracellular and/or intracellular Fe3+ ions. PMID- 9202180 TI - The rotational diffusion of LH receptors differs when receptors are occupied by hCG versus LH and is increased by cytochalasin D. AB - We have examined the rotational diffusion of the luteinizing hormone (LH) receptors binding human chorionic gonadotropin (hCG) or ovine luteinizing hormone (oLH) in MA-10 Leydig tumor cells using time-resolved phosphorescence anisotropy techniques. LH receptors binding erythrosin isothiocyanate (ErITC)-derivatized oLH were rotationally mobile with rotational correlation times of 62 micros, 48 micros, 38 micros, and 29 micros at 4 degrees C, 15 degrees C, 25 degrees C, and 37 degrees C, respectively. ErITC-hCG bound to the LH receptor was rotationally immobile, showing no anisotropy decay at 4 degrees C, 15 degrees C, 25 degrees C, and 37 degrees C. To determine whether cytoskeletal components influenced the rotational diffusion of LH receptors, we measured rotational diffusion of LH receptors on MA-10 cells treated with 20 microg/ml cytochalasin D and on plasma membrane preparations. Following 1 h exposure to cytochalasin D, the rotational correlation times for hCG-occupied LH receptors were typically 11 micros at 37 degrees C compared to > 1000 micros on untreated cells. Treatment of MA-10 cells with cytochalasin B or colchicine had no affect on LH receptor rotational diffusion. Rotational correlation times for LH-occupied receptors decreased from 29 micros to 12 micros at 37 degrees C following cytochalasin D treatment. The rotational diffusion of LH receptors on plasma membrane preparations was similar to that observed for LH- and hCG-occupied receptors on intact cells treated with cytochalasin D. These various results indicate that there are differential effects of LH and hCG binding on the interactions of LH receptors with plasma membrane proteins and that microfilaments anchor the hCG- and LH-occupied receptors. PMID- 9202181 TI - Retinol free and retinol complexed beta-lactoglobulin binding sites in bovine germ cells. AB - A high affinity specific binding site for bovine beta-lactoglobulin (BLG) was identified in bovine germ cell plasma membrane enriched fractions. Binding was found to be reversible and pH-dependent with maximum binding occurring at pH 5. The on-rate and off-rate constants were 2.26 +/- 0.8 x 10(5) M(-1) min(-1) (n = 3) and 0.016 +/- 0.004 min(-1) (n = 3), respectively. Scatchard analysis showed a single class of binding sites, with 12.38 +/- 4.62 x 10(12) sites per mg of membrane protein (n = 3) and a dissociation constant (K(D)) estimated at 26.43 +/ 2.68 nM. There was inhibition of iodinated-BLG (variant A) (125I-BLGA) binding to germ cell plasma membrane enriched fractions in the presence of unlabelled BLG variant A, BLG variant B, retinol complexed BLGA and human retinol-binding protein. Inhibition was observed neither with BSA nor with lactoferrin. 125I-BLGA incubated with a Triton X-100 solubilized plasma membrane fraction formed a high molecular mass complex in Superose 12B gel filtration. This receptor complex disappeared in the presence of unlabelled BLGA and in the presence of 10 mM EDTA. The results suggest that germ cell plasma membrane may contain a receptor which is capable of binding either retinol free or retinol complexed BLGA. PMID- 9202182 TI - tTGase/G alpha h protein expression inhibits adenylate cyclase activity in Balb-C 3T3 fibroblasts membranes. AB - Stably transfected Balb-C 3T3 fibroblasts (clone 5), overexpressing a catalytically active tissue transglutaminase, showed a basal adenylate cyclase activity lower than control cells (clone 1). Several modulators of the adenylate cyclase activity (forskolin, Mn2+ and pertussis toxin) showed the existence of a marked negative control on the adenylate cyclase activity present in clone 5 cells. Very interestingly, this same marked negative control was also found in a Balb-C 3T3 fibroblast clone stably transfected with a mutagenized human tissue transglutaminase (mut277 cys > ser) virtually devoid of transglutaminase catalytic activity (clone Ser). Conversely, a significant increase of the adenylate cyclase activity was observed in bovine aortic endothelial cells after the lowering of tissue transglutaminase expression levels by the transfection of an eukaryotic expression vector containing the gene for tissue transglutaminase in antisense orientation. All these findings suggest a possible role for type II tissue transglutaminase as a negative modulator of the adenylate cyclase activity in different cell types, beside its transglutaminase enzyme activity. PMID- 9202183 TI - Effects of calmodulin antagonists on calcium pump and cytosolic calcium level in human neutrophils. AB - The concentration of cytosolic free calcium was monitored in suspensions of intact human neutrophils in phosphate-buffered saline by means of the fluorescent indicator Indo 1 trapped in the cytosol. Trifluoperazine and n-(6-aminohexyl)-5 chloro-1-naphthalenesulfonamide markedly reduced the amplitude of the transient increase in cytosolic Ca2+ triggered by CaCl2 as well as by N-formyl-methionyl leucyl-phenylalanine. The effect of the calmodulin antagonists on the calcium burst observed upon cell activation was much more pronounced in the presence of extracellular free calcium than in EGTA-containing media; it was not inhibited by wortmannin or thapsigargin. Nevertheless, trifluoperazine and n-(6-aminohexyl)-5 chloro-1-naphthalenesulfonamide inhibited the plasma-membrane Ca2+ ATPase if added to plasma membrane-enriched fractions of neutrophils. These results suggest that calmodulin antagonists affect calcium ion influx even if they inhibit plasma membrane Ca2+ ATPase. PMID- 9202184 TI - Characterization of the in vitro synthesized arabinan of mycobacterial cell walls. AB - Previous studies have shown that polymerized [14C]arabinan can be synthesized from polyprenylphosphate-[14C]arabinose by the particulate enzymes of Mycobacterium smegmatis [R.E. Lee, K. Mikusova, P.J. Brennan and G.S Besra (1995) J. Am. Chem. Soc. 117, 11829-11832]. In the present investigation, the [14C]arabinan product was biochemically characterized. Sizing chromatography revealed a molecular weight consistent with that expected from mature arabinan. Digestion of the [14C]arabinan with a mixture of arabinases produced oligo[14C]arabinoside fragments including hexa[14C]arabinoside and tetra[14C]arabinoside which originated from the non-reducing terminal regions of the polymer, and di[14C]arabinoside from the internal regions of the polymer. These arabinoside fragments represent the major known structural motifs that comprise the arabinan segment of arabinogalactan and lipoarabinomannan. The presence of [14C]arabinose in both the internal and external regions of the [14C]arabinan suggests that polyprenylphosphate-arabinose is the major, and perhaps the only, donor of arabinosyl residues in mycobacteria. PMID- 9202185 TI - Heterologous Kluyveromyces lactis beta-galactosidase production and release by Saccharomyces cerevisiae osmotic-remedial thermosensitive autolytic mutants. AB - The beta-galactosidase from Kluyveromyces lactis is a high molecular weight protein with commercial interest. A major drawback of its industrial production is the high cost associated with extraction and downstream processing due to its intracellular nature. In this work, the effectiveness of the utilization of Saccharomyces cerevisiae LD1 and LHDP1 strains, osmotic-remedial mutants which lyse at 37 degrees C, for the heterologous production and release into the extracellular medium of this protein has been proved. The highest absolute values of released beta-galactosidase have been obtained with the protease-deficient strain LHDP1 by osmotic shock. PMID- 9202186 TI - The origin of an EPR signal observed in dithiocarbamate-loaded tissues. Copper(II)-dithiocarbamate complexes account for the narrow hyperfine lines. AB - We examined the electron paramagnetic resonance spectra of some copper(II) dithiocarbamate complexes in various media at liquid nitrogen temperature. It is demonstrated that Cu(II)-dithiocarbamate complexes exhibit atypical spectra with narrow hyperfine splitting, and that the dithocarbamates forming a water insoluble Cu(II) complex are permeable in tissues, while those yielding water soluble complexes are not. PMID- 9202187 TI - 2-Deoxyecdysone is a circulating ecdysteroid in the beetle Zophobas atratus. AB - A qualitative analysis of ecdysteroids has been performed during the post embryonic development of the tenebrionid beetle, Zophobas atratus, by high performance liquid chromatography (HPLC) combined with enzyme immunoassay (EIA) using two different antibodies. Three HPLC peaks were found to be immunoreactive, in hemolymph extracts of both sexes. Moreover, these peaks had ecdysteroid-like UV spectra, determined using a photodiode array detector. The use of two different HPLC systems (reverse and normal phases), in combination with two different EIA antibodies, allowed us to identify 20-hydroxyecdysone (20E) and ecdysone (E), as the two main ecdysteroids, but also suggested the presence of 2 deoxyecdysone (2dE) as the third hemolymph component. Secretion of putative 2dE, together with E (but not 20E) was also demonstrated in vitro from incubations of prothoracic glands and of tegumental explants. In these experiments, either in vivo or in vitro, 3-dehydroecdysone was never observed. Our observations thus strongly suggest that 2dE is a circulating ecdysteroid in Z. atratus and may function as a prohormone during the development of some insects. PMID- 9202188 TI - Optical and electron paramagnetic resonance absorption spectra of complexes of nitric oxide synthase I with isocyanides. AB - Neuronal nitric oxide synthase (NOS I) was purified from porcine brains, and optical and EPR spectra of the complexes of NOS I with isocyanides were investigated. The complex of oxidized NOS I with tert-butylisocyanide exhibited optical absorption peaks at 437 and 560 nm in the difference spectrum, whereas with tert-butylisocyanide and phenylisocyanide, optically reduced NOS I isocyanide complexes with absorbance maxima at 433, 451, 541 and 573 nm were produced. The dissociation constants of the NOS I-isocyanide complexes were optically determined, the constants being significantly larger than those of microsomal cytochromes P-450. Phenylisocyanide did not affect the optical spectrum of oxidized NOS I. A high concentration of phenylisocyanide also had no effect on the EPR spectrum of oxidized NOS I. The optical spectra of the reduced NOS I-isocyanide complexes were pH-dependent. With increasing pH, the intensity of the absorbance at 451 nm of the complexes increased and that of the absorbance at 433 nm decreased in parallel. Upon the addition of a saturating concentration of L-arginine, the difference spectra of the reduced NOS I-phenylisocyanides complex showed a drastic change, i.e., an increase in optical intensity at 433 nm and a concomitant decrease in the intensity at 451 nm. In titration experiments with L-arginine, spectral binding, Ks = 2.5 microM, was determined from the absorbance increase at 433 nm. No spectral change was observed on the addition of the same concentration of D-arginine. N(omega)-nitro-L-arginine methyl ester (NAME), a potent inhibitor of NOS I, had a similar effect to L-arginine, but the time course of the spectral change was very slow. These results suggest that: (1) the heme-iron pocket of NOS I will be narrower than those of the microsomal and mitochondrial cytochromes P-450; and (2) the binding of L-arginine and its analogue to their binding sites caused conformational changes around the ferrous heme moiety of NOS I. PMID- 9202189 TI - Effect of glycosylation on the stability of alpha1-antitrypsin toward urea denaturation and thermal deactivation. AB - The effects of glycosylation on the stability of human alpha1-antitrypsin were investigated. The transition midpoints in urea-induced equilibrium unfolding of a non-glycosylated recombinant, a yeast version of glycosylated, and human plasma alpha1-antitrypsin were 1.8 M, 2.2 M, and 2.5 M at 25 degrees C, respectively. Kinetic analyses of unfolding and refolding revealed that glycosylation retarded the unfolding without affecting the refolding rate significantly, suggesting that the stability increase is due to the stabilization of the native state as opposed to the destabilization of the unfolded state. In thermal deactivation, which is a heat-induced aggregation process, the unglycosylated recombinant alpha1 antitrypsin was deactivated most easily, which was followed in order by the yeast, and the plasma form. The results indicate that glycosylation confers the increase in stability of alpha1-antitrypsin, and that the oligomannose sugars present on the yeast form produce a less stable molecule than the complex type sugars on the plasma form. It appears that the effect of glycosylation on the enhancement of thermal resistance is exerted through the increase in conformational stability. However, a stable recombinant variant (Phe 51 --> Cys) that showed the same conformational stability as the plasma form was less resistant to thermal denaturation than the plasma alpha1-antitrypsin. The results suggest that the existence of carbohydrate moiety per se as well as the conformational stability contribute to the kinetic stability of alpha1 antitrypsin toward aggregation. PMID- 9202190 TI - Possible role of isoaspartyl methyltransferase towards regulation of acid trehalase activity in Saccharomyces cerevisiae. AB - Logarithmically growing cells of S. cerevisiae contained high neutral trehalase (NT) activity while stationary-phase cells had high acid trehalase (AT) activity. Change in activity profile of AT and NT were different during growth under different conditions, particularly during growth in acetate medium and up to 1 h of germination period, but that for AT and isoaspartyl methyltransferase (IMT) were found to be almost identical. Concomitant increase in NT activity as well as increase in cAMP level was noticed at the onset of spore germination. Increase in AT and IMT activities as well as decrease in S-adenosyl-L-methionine (AdoMet) level were noticed during stationary phase of growth. Acidic polyacrylamide gel electrophoresis and subsequent autoradiography revealed that substrate of IMT was a protein of molar mass around 82 kDa which could be an AT. Methylated AT was found to be more active while non-methylated AT was relatively less active in comparison to the untreated sample. Since AT existed as an equilibrium mixture of protomer and oligomer, it was suggested that IMT catalysed carboxyl methylation might have some contribution towards the regulation of AT activity. PMID- 9202191 TI - Effects of magnesium and temperature on the conformation and reassociation of Escherichia coli and Sulfolobus solfataricus ribosomes. AB - The structural response of the ribosomes of the extremely thermophilic archaeon Sulfolobus solfataricus was analysed and compared to that of the mesophilic (E. coli) ribosomes by assaying ethidium bromide (EB) binding to the native 70S particles as a function of magnesium concentration. We found that the thermophilic ribosomes bound more EB than their mesophilic counterparts; on the other hand, inhibition of EB binding by Mg2+ ions was more effective in the E. coli 70S particle. In Sulfolobus, the separated 30S and 50S subunits and the 70S particle bound the drug in a similar fashion, whereas the E. coli 70S had a reduced number of binding sites with respect to the subunits. Light scattering measurements as a function of Mg2+ concentration were carried out at various temperatures to study the interaction between the ribosomal subunits from the thermophilic and the mesophilic bacteria. As expected, the association of ribosomal subunits in E. coli was magnesium dependent and could be observed also at low temperature. By contrast, the interaction between Sulfolobus ribosomal subunits was obligatorily dependent upon both magnesium ions and a temperature of at least 80 degrees C, close to the physiological optimum for cell growth (87 degrees C). PMID- 9202192 TI - In vivo assessment of metabolic perturbations following alanine and glucagon administration using 31P-MRS in the rat. AB - This study set out to validate the use of 31P-NMR spectroscopy together with alanine +/- glucagon infusions to assess hepatic gluconeogenic flux in vivo. Bolus infusions of alanine (2.8 or 5.6 mmol/kg) +/- glucagon (250 microg/kg) were used. Maximal changes in the phosphomonoesters (PME), inorganic phosphate (Pi) and beta-NTP occurred 40 mins post infusion. PME increased 13.1% (p < 0.02) and 20.8% (P < 0.01) at 2.8 mmol/kg + glucagon and 5.6 mmol/kg +/- glucagon, respectively. Pi was unaltered at 2.8 mmol/kg but increased by 28.8% (P < 0.01) at 5.6 mmol/kg alanine + glucagon. beta-NTP decreased by 14.4% (P < 0.02) and 16.1% (P < 0.02) at 5.6 mmol/kg -/+ glucagon, respectively. This latter infusion showed slower recovery rates of NTP which remained 12.3% (P < 0.05) lower 70 min post infusion compared with pre-infusion values. 31 P-NMR analysis of liver extracts revealed that PME increases were partly due to 3-phosphoglycerate and corroborated reductions in beta-NTP and gamma-NTP: beta-NDP ratio upon infusion of 5.6 mmol/kg alanine +/- glucagon. Hepatic glucose output from perfused liver experiments showed no difference between alanine concentrations indicating maximal glucose output at the lower concentration. This study has shown that in vivo 31P-NMR in combination with alanine infusion, can be used to determine metabolic changes associated with gluconeogenesis. PMID- 9202193 TI - Preparation and ESR spectroscopic characterization of the zinc(II) and cadmium(II) complexes of streptonigrin semiquinone. AB - Semiquinone metal complexes derived from the antitumor antibiotic streptonigrin have been prepared for the first time. They were obtained by reduction of the zinc(II) and cadmium(II) complexes of the parent aminoquinone ligand with sodium borohydride, followed by air oxidation of the intermediate dihydroquinones. Alternatively, N-benzyldihydronicotinamide reduction was used to produce the same Cd(II) complex of the p-semiquinone free radical. Electron spin resonance spectroscopic studies showed that metal binding significantly changes the spin densities of the unpaired electron which is confined to the quinolinesemiquinone moiety of the complexed antibiotic. Complexation with both Cd(II) and Zn(II) perturbs the coupling constants of all atoms involved in delocalization of the unpaired electron, shifting its distribution toward the pyridine ring. The coupling constant of the pyridine ring-proton adjacent to the semiquinone ring increases from 0.31 to 0.43 G in the Cd(II) complex in methanol, while the proton meta to the pyridine nitrogen increases from 1.76 to 1.96 G. Furthermore, the coupling constant of the heterocyclic nitrogen increases from 0.46 to 0.61 G. A similar trend is noted for the Zn(II) complex as well, including the observed decrease in splitting constant of the amino nitrogen from 1.34 to 1.08 G, and perturbation of the previously equivalent amino protons from 0.89 and 0.89 to 1.09 and 0.95 G. The spectral parameters have been confirmed by deuteration. Complexation studies using isotopically enriched 113Cd(II) revealed hyperfine coupling of the unpaired electron of the p-semiquinone and the nuclear spin of 113Cd(II), indicating direct coordination between the metal and the complexing ligand. Although the metal complexes could readily be prepared in a series of different solvent systems, they appear to have substantially shorter half lives than the non complexed p-semiquinone radical (5 to 15 min vs. 2 to 3 wk in sealed ampoules). Formation of tight-binding p-semiquinone metal complexes as here described should provide useful leads for the design of related systems to study p-quinone-metal interactions for mechanistic elucidation of metal ion catalyzed quinone-dependent electron transfer reactions in biological oxidations. PMID- 9202194 TI - Temperature-induced phase partitioning of peptides in water solutions of ethylene oxide and propylene oxide random copolymers. AB - A thermoseparating random copolymer (Ucon 50-HB-5100) composed of (50%) ethylene oxide and (50%) propylene oxide has been used to form an aqueous two-phase system by heating the polymer-water solution above the cloud point of the copolymer. In the formed two-phase system a water rich top phase is in equilibrium with an aqueous polymer rich bottom phase. The partitioning of amino acids and peptides in this aqueous two-phase system has been studied. Hydrophobic peptides (containing aromatic amino acids) were strongly partitioned to the polymer rich phase, while hydrophilic peptides were enriched in the water rich phase. The effect of temperature on the partitioning was investigated and a decreased partitioning to the polymer rich phase was obtained upon temperature increase. The effect of two salts (NaClO4 and Na2SO4) on the partitioning of a positively charged polypeptide, poly(Lys, Trp), was very strong. With NaClO4 the polypeptide was quantitatively partitioned to the polymer rich phase while with Na2SO4 the polypeptide was partitioned to the water rich phase. Model calculations based on a modified Flory-Huggins theory have been performed to better understand the experimental behavior. PMID- 9202195 TI - Enzymic synthesis of 3'-O- and 6'-O-N-acetylglucosaminyl-N-acetyllactosaminide glycosides catalyzed by beta-N-acetyl-D-hexosaminidase from Nocardia orientalis. AB - beta-N-acetyl-D-hexosaminidase from Nocardia orientalis catalyzed the synthesis of beta-D-GlcNAc-(1 --> 3)-beta-D-Gal-(1 --> 4)-beta-D-GlcNAc-OC6H4NO2-p (1) and beta-D-GlcNAc-(1 --> 6)-beta-D-Gal-(1 --> 4)-beta-D-GlcNAc-OC6H4NO2-p (2) with its isomer beta-D-Gal-(1 --> 4)-[beta-D-GlcNAc-(1 --> 6)]-beta-D-GlcNAc-OC6H4NO2 p (3) through N-acetylglucosaminyl transfer from N-,N'-diacetylchitobiose to p nitrophenyl beta-N-acetyllactosaminide. The enzyme formed a mixture of trisaccharides 1, 2, and 3 in a ratio of 11:33:56. In the case, when an inclusion complex of p-nitrophenyl beta-N-acetyllactosaminide with alpha-CD was used, compounds 1, 2, and 3 were formed in a molar ratio of 24:63:13. The regioselectivity of glycosidase-catalyzed formation of the trisaccharide glycosides was substantially changed. It resulted not only in a significant increase of the proportion of the desired compounds 1 and 2 but also in the substantial increase of the overall yield of transfer products. PMID- 9202196 TI - Antioxidant activity of polyphenolics in diets. Rate constants of reactions of chlorogenic acid and caffeic acid with reactive species of oxygen and nitrogen. AB - Phenolic non-flavonoid compounds in diets, such as chlorogenic acid and caffeic acid are widely recognized to be antioxidants. However, it is not known how these phenolics scavenge reactive species of oxygen and nitrogen. We determined the rate constants of the reactions between the phenolics with superoxide and hydroxyl radical with a pulse radiolysis. The second-order rate constants of the reactions of chlorogenic acid with superoxide and hydroxyl radical were 1.67 +/- 0.14 x 10(6) M(-1) s(-1) and 3.34 +/- 0.19 x 10(9) M(-1) s(-1), respectively, while those of caffeic acid with superoxide and hydroxyl radical were 0.96 +/- 0.01 x 10(6) M(-1) s(-1) and 3.24 +/- 0.12 x 10(9) M(-1) s(-1), respectively. By scavenging peroxy radical chlorogenic acid inhibited the initiation of chain lipid peroxidations by organic free radical. The second-order rate constant of the reaction of chlorogenic acid with peroxy radical was estimated to be 1.28 +/- 0.11 x 10(5) M(-1) s(-1). Chlorogenic acid was rapidly oxidized by peroxynitrite in concentration- and pH-dependent manners and its rate constant was determined to be 1.6 +/- 0.7 x 10(5) M(-1) s(-1), using competitive inhibitions by glutathione and methionine. PMID- 9202197 TI - Phenylenediamine induced hepatocyte cytotoxicity redox. Cycling mediated oxidative stress without oxygen activation. AB - Muscle necrosis induced by various phenylenediamine derivatives has been correlated with their autoxidation rate. However, a more detailed investigation of the cytotoxic mechanism using a model system of isolated hepatocytes and 2,3,5,6-tetramethylphenylenediamine (DD) shows little oxygen activation as indicated by the absence of cyanide resistant respiration, lipid peroxidation and lack of cytoprotection by iron chelators, superoxide dismutase mimics and xanthine oxidase inhibitors. Cytotoxicity was however attributed to oxidative stress as GSH was not only rapidly oxidized to GSSG but mixed protein disulfide formation also occurred. Furthermore, the disulfide reductant dithiothreitol added some time after DD restored protein thiols and prevented further cytotoxicity. This oxidative stress was attributed to a futile two electron redox cycle involving oxidation of DD to the corresponding diimine by the mitochondrial electron transport chain and rereduction by DT diaphorase. Evidence suggesting this was that both diimine accumulation and the ensuing cytotoxicity were markedly increased by inactivating hepatocyte DT diaphorase but were prevented by a subtoxic concentration of the mitochondrial respiratory inhibitor cyanide. Furthermore, addition of NADH generating substrates such as lactate, sorbitol, xylitol or ethanol prevented DD induced GSH oxidation and cytotoxicity. This suggests that DD undergoes intracellular redox cycling without oxygen activation until the hepatocyte is unable to maintain redox homeostasis and mixed protein disulfide cytotoxicity ensues. PMID- 9202198 TI - Localization of renal Cu-binding metallothionein induced by Au injection into rats. AB - The localization of Au-induced metallothionein (MT) in kidneys is reported. Au, Cu and Zn contents in kidneys and liver increased after Au injection. Especially the Cu content in the kidney increased in comparison with the Zn content. The yellow-orange autofluorescent signals which are a marker of Cu-MT were observed predominantly in the outer stripe of the outer medulla with a ring shape in the kidneys of Au-injected rats. MT mRNA was also located in only the outer stripe of the outer medulla. Neither autofluorescence nor MT mRNA was found in the kidneys of control rats. These results indicate that MT was biosynthesized in only the outer stripe of the outer medulla and the biosynthesized MT was bound to Cu. Representative Sephadex G-75 elution profiles of the renal cytosol of rats injected with Au showed that Au, Cu and Zn contents in MT fractions increased after Au injection. Interestingly, Cu in MT fractions dramatically increased in comparison with Zn in the MT fractions in spite of Au injection into rats. Only the Cu-containing MT fractions emitted a yellow-orange autofluorescence. The accumulated Cu in the kidneys of Au-injected rat was thought to be associated with renal toxicity. PMID- 9202199 TI - Growth inhibition by insulin-like growth factor (IGF) binding protein-3--what's IGF got to do with it? PMID- 9202200 TI - Follicle-stimulating hormone induces terminal differentiation in a predifferentiated rat granulosa cell line (ROG). AB - The divergent commitment of ovarian granulosa cells to either proliferation and differentiation or programmed cell death directly reflects the process of follicular dominance and atresia. This process is regulated by FSH and local paracrine factors. To further analyze the role of FSH and intraovarian factors in follicular selection, we have established a rat ovarian granulosa (ROG) cell line from prepubertal (p14) rats. ROG cells are cultured in serum-free medium with activin A, but without FSH. ROG cells bind FSH and respond to FSH by a burst of cell proliferation and increased progesterone secretion. These results support the hypothesis that activin, but not FSH, is an important factor in the maintenance of immature granulosa cells. After exposure of ROG cells to FSH, withdrawal of FSH from the cultures results in apoptotic cell death. ROG cells start active membrane blebbing by 2 h after FSH withdrawal, and most cells die within 7 h. Thus, FSH-induced ROG cells differentiate into a more mature granulosa phenotype, which is nonmitotic and dependent on FSH for survival. The ROG cell line may thus provide a good in vitro model of follicular selection. PMID- 9202201 TI - A novel calcium-activated apamin-insensitive potassium current in pituitary gonadotrophs. AB - In cultured rat pituitary gonadotrophs, GnRH-induced oscillations in cytosolic calcium concentration ([Ca2+]i) are associated with periodic membrane hyperpolarization. The hyperpolarizing waves are secondary to the activation of apamin-sensitive Ca2+-activated K+ channels that account for a single class of 125I-apamin binding sites present in these cells. In a substantial fraction of gonadotrophs, however, we observed a Ca2+-controlled oscillatory current that was resistant to apamin, even at concentrations five orders of magnitude higher than the dissociation constant (Kd) observed in the binding experiments. With the K+ in the pipette, the apamin-resistant current showed a reversal potential of -42 mV, nearly 40 mV more positive than that of the apamin-sensitive current. With Cs+ in place of K+ in the pipette solution, both the size of the apamin insensitive current and its reversal potential remained unchanged. Ion substitution studies further revealed that the reversal potential was independent of Cl-. In contrast, an 11 mV hyperpolarizing shift in the reversal potential occurred when extracellular Na+ was reduced to 80 mM. In cells expressing apamin resistant conductances, addition of apamin evoked a marked increase in the duration of the action potentials and reduction in the frequency of spontaneous spiking. In the presence of GnRH, gonadotrophs exhibit the typical burst pattern of electrical activity. Further exposure of the cells to apamin depolarized the membrane from a silent phase bursting level of about -80 mV to a new level of about -40 mV. These observations indicate that, in addition to apamin-sensitive current, a subpopulation of pituitary gonadotrophs also expresses a cationic component of the Ca2+-activated membrane conductance that has the potential to remodulate spontaneous and agonist-induced electrical activity. PMID- 9202202 TI - Parathyroid hormone-related protein is induced in the adult liver during endotoxemia and stimulates the hepatic acute phase response. AB - Previously, we reported that PTH-related protein (PTHrP) gene expression is induced in vital organs, including the liver, during endotoxemia. The liver plays a central role in the acute phase response (APR), a cytokine-mediated host defense against infection and inflammation that includes increased production of acute phase proteins and lipids by hepatocytes. Because PTHrP is thought to act locally at its site of production, in vivo studies were carried out to determine whether PTHrP could contribute to the induction of the hepatic APR. Hepatic PTHrP messenger RNA (mRNA) levels were induced acutely in rat liver in response to a near lethal dose of endotoxin. PTHrP protein, which was located by immunohistochemical staining in hepatocytes from both control and LPS-treated rats, was markedly induced in periportal hepatocytes in response to LPS treatment. Co-incident with this transient increase in PTHrP gene expression, PTH/PTHrP receptor mRNA levels were down-regulated. Administration of PTHrP(1 34), a PTH/PTHrP receptor agonist, to mice increased hepatic serum amyloid A (SAA) mRNA levels as well as circulating levels of SAA. In addition, PTHrP(1-34) increased serum triglyceride (TG) levels in rats and mice in a dose-dependent fashion. The hypertriglyceridemic effect of PTHrP(1-34) was accompanied by an increase in hepatic fatty acid synthesis. In contrast, PTHrP(7-34) amide, a receptor antagonist, had no effect on serum SAA or TG levels. These results, which provide evidence for the regulated expression of PTHrP in adult liver, suggest that PTHrP may be one additional member of the cytokine cascade produced locally in liver that can act to stimulate the hepatic acute phase response. PMID- 9202203 TI - Acute and chronic exposure to tumor necrosis factor-alpha fails to affect insulin stimulated glucose metabolism of isolated rat soleus muscle. AB - To better understand the effects of tumor necrosis factor-alpha (TNF alpha) on insulin sensitivity, direct interaction of the peptide with freshly isolated rat soleus muscle strips was investigated. Muscles were exposed to TNF alpha at concentrations ranging from 0.01-5 nmol/liter. Rates of insulin-stimulated (5 or 100 nmol/liter) glucose metabolism were determined after periods of TNF alpha preexposure of 30 min, 6 h, and 24 h. Independent of exposure time, TNF alpha failed to exert any significant effect on rates of 3H-2-deoxy-glucose transport (stimulation by 100 nmol/liter insulin after preincubation without vs. with 5 nmol/liter TNF alpha, cpm/mg x h: 30 min, 779 +/- 29 vs. 725 +/- 29; 6 h, 652 +/- 56 vs. 617 +/- 60; 24 h, 911 +/- 47 vs. 936 +/- 31) or glucose incorporation into glycogen (micromol/g x h: 30 min, 5.19 +/- 0.22 vs. 5.25 +/- 0.41; 6 h, 2.08 +/- 0.10 vs. 2.09 +/- 0.17; 24 h, 2.51 +/- 0.21 vs. 2.41 +/- 0.26). In parallel, TNF alpha neither affected insulin-stimulated rates of glucose oxidation (CO2 production) and anaerobic glycolysis (lactate release), nor muscle glycogen content. In conclusion, these findings do not support the hypothesis of muscle insulin desensitization by TNF alpha via autocrine or paracrine mechanisms. The obtained data favor the concept that TNF alpha-dependent muscle insulin resistance in vivo depends on indirect effects rather than direct interaction of the peptide with skeletal muscle. PMID- 9202204 TI - Interleukin (IL)-1beta increases glucose uptake and induces glycolysis in aerobically cultured rat ovarian cells: evidence that IL-1beta may mediate the gonadotropin-induced midcycle metabolic shift. AB - This communication explores the possibility that interleukin (IL)-1beta, a putative intermediary in the ovulatory process, may take part in the gonadotropin driven midcycle diversion of ovarian carbohydrate metabolism toward glycolysis. We examined the effect of treatment with IL-1beta on glucose metabolism in aerobically cultured whole ovarian dispersates from immature rats. Treatment with IL-1beta increased cellular glucose consumption/uptake, stimulated extracellular lactate accumulation and media acidification, and decreased extracellular pyruvate accumulation in a receptor-mediated, time-, dose- and cell density dependent manner. Endogenous IL-1beta-like bioactivity was shown to mediate the ability of gonadotropins to exert these same metabolic effects. The IL-1beta effect was also (1) apparent over a broad range of glucose concentrations, inclusive of the putative physiological window; (2) relatively specific, because tumor necrosis factor-alpha and insulin were inactive; (3) contingent upon cell cell cooperation (4) and reliant on de novo protein synthesis. Comparison of the molar ratios of lactate accumulation to glucose consumption in IL-1beta-replete vs. IL-1beta-deplete cultures suggests that IL-beta promotes the conversion of all available glucose to lactate but that other substrates for lactate production may also exist. However, no lactate was generated by cells grown under glucose free conditions. Taken together, our data suggest that IL-1beta may act as a metabolic hormone in the ovary. Subject to the limitations of the in vitro paradigm, our data also suggest that IL-1beta may mediate the gonadotropin associated midcycle shift in ovarian carbohydrate metabolism. By converting the somatic ovarian cells into a glucose-consuming glycolytic machinery, IL-1beta may establish glycolysis as the main energy source of the relatively hypoxic preovulatory follicle and the resultant cumulus-oocyte complex. The consequent oxygen sparing may conserve the limited supply of oxygen needed for vital biosynthetic processes such as steroidogenesis. This adaptational response may also provide the glycolytically incompetent oocyte with the obligatory tricarboxylic cycle precursors it depends on to meet the increased energy demands imposed upon it by the resumption of meiosis. PMID- 9202205 TI - Alternative signaling mechanism of leukemia inhibitory factor responsiveness in a differentiating embryonal carcinoma cell. AB - Leukemia inhibitory factor (LIF) is a cytokine that plays an important role during mouse embryogenesis. We showed that adenovirus E1A represses the interleukin-6 signal transduction pathway that uses the same JAK tyrosine kinase and STAT (signal transducer and activator of transcription) transcription factor as LIF. Here, we report that the LIF-JAK-STAT signal transduction pathway is blocked in cellular E1A-expressing undifferentiated F9 cells, and that the block is overcome by retinoic acid-induced differentiation. LIF failed to stimulate the expression of the acute phase response element (APRE)-driven luciferase gene in undifferentiated F9 cells, whereas the luciferase activity was remarkably increased by LIF treatment in differentiated F9 (dF9) cells. We analyzed the mechanism of the APRE regulation and found that the LIF-induced APRE-binding activity was regulated in a differentiation-dependent manner. The protein levels and the tyrosine phosphorylation of JAK1, JAK2, and STAT3 in F9 cells were not different from those in dF9 cells. The exogenous expression of activated c-Ha-ras partially recovered the LIF responsiveness of the APRE-luciferase gene in F9 cells, but the dominant negative ras N-17 did not repress the LIF-induced activation of APRE-luciferase in dF9 cells. These results suggested that an unknown coactivation process that is partially compensated by Ras is required for STAT3-APRE binding in F9 cells. PMID- 9202206 TI - Adrenalectomy after weaning restores beta3-adrenergic receptor expression in white adipocytes from C57BL/6J-ob/ob mice. AB - The role of hypercorticism in the development of compromised beta-adrenergic signaling in adipose tissue was assessed in ob/ob mice adrenalectomized at 4 weeks of age and studied 1 and 3 weeks thereafter. Adrenalectomy prevented the rapid increase in body weight and fat deposition between 4 and 5 weeks of age in ob/ob mice and produced a phenotype indistinguishable from that of lean mice. However, adrenalectomized ob/ob mice became intermediate between lean and ob/ob mice by 7 weeks of age. Adipocyte beta3-adrenergic receptor (AR) messenger RNA levels were similar between lean and adrenalectomized ob/ob mice at both time points and were 4- to 8-fold higher than messenger RNA levels in ob/ob mice. As judged by maximal activation of adenylyl cyclase by a beta3-AR-selective agonist, adrenalectomy also restored functional activity of the beta3-AR to levels above or equivalent to those seen in lean mice at both time points. The present results suggest that development of hypercorticism at or before weaning in ob/ob mice represses expression of the beta3-AR and prevents the normal postweaning development of this signaling system in the adipocyte. PMID- 9202207 TI - Inhibition of fetal adrenal adrenocorticotropin receptor messenger ribonucleic acid expression by betamethasone administration to the baboon fetus in late gestation. AB - Throughout the majority of intrauterine development, the primate fetal adrenal gland is comprised primarily of fetal zone cells and only late in gestation do definitive zone cells, which express the enzyme delta5-3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD) emerge to produce cortisol. The present study was designed to determine whether the induction of definitive zone ACTH receptor messenger RNA (mRNA) levels and components of the steroidogenic pathway known to be expressed specifically in the definitive zone, e.g. the 3beta-HSD enzyme, are dependent upon fetal pituitary ACTH. Fetal pituitaries and adrenal glands were obtained on day 165 (term = day 184) from untreated controls (n = 7) and from baboons in which betamethasone was administered im to the fetus (0.6 mg/100 microl; n = 4) or to the fetus (0.6 mg) and mother (6 mg/ml; n = 4) every other day between days 150 and 164 of gestation. Although fetal pituitary weight was not altered by betamethasone, POMC mRNA levels determined by in situ hybridization were lower (P < 0.05) in betamethasone-treated (0.34 +/- 0.07 arbitrary densitometric units) than in untreated controls (0.63 +/- 0.04). Associated with this decline in pituitary POMC, levels of the major 3.4-kb mRNA transcript for the ACTH receptor expressed as a ratio of beta-actin were approximately 80% lower (P < 0.05) in fetal adrenals of betamethasone-treated baboons (0.12 +/- 0.02) than in untreated controls (0.84 +/- 0.05). In situ hybridization indicated that ACTH receptor mRNA expression in the definitive zone exceeded that in the fetal zone and was reduced by betamethasone. Associated with the decrease in ACTH receptor expression, fetal adrenal weight was suppressed (P < 0.05) by 50% and reflected a marked reduction (P < 0.05) in the size of the cells of the definitive and fetal zones. Betamethasone treatment also induced a decrease (P < 0.05) in the width (microm) of the definitive zone (183 +/- 14 vs. 128 +/- 7; determined by immunohistochemical expression of 3beta-HSD), as well as the levels of the mRNA and protein for 3beta-HSD. Levels of the mRNA for the LDL receptor and the enzymes 17alpha-hydroxylase-C(17,20) lyase and P450 cholesterol side chain cleavage were also suppressed in adrenals of betamethasone-treated baboons. These findings indicate that treatment of the baboon fetus with betamethasone in late gestation suppressed fetal pituitary POMC mRNA expression and ACTH receptor mRNA levels in the fetal adrenal gland, as well as the hypertrophy and ACTH receptor mRNA and 3beta-HSD mRNA/protein levels in the cells comprising the newly emerging definitive zone. We conclude that ACTH is necessary for the up-regulation of the mRNAs for the ACTH receptor and steroidogenic enzymes in the definitive zone of the primate fetal adrenal gland in late gestation. PMID- 9202208 TI - Neonatal hypothyroidism permanently alters follicle-stimulating hormone and luteinizing hormone production in the male rat. AB - Transient neonatal hypothyroidism, induced with the goitrogen 6-n-propyl-2 thiouracil (PTU), results in dramatic increases in both testis size and sperm production in the adult rat. The observed increases in testis size and function occur in the presence of normal circulating testosterone levels. However, circulating gonadotropin levels are chronically reduced by 30-50% at all times in treated males. To better understand the permanent reduction in serum gonadotropin levels following transient neonatal hypothyroidism, we conducted a series of experiments to evaluate pituitary and hypothalamic function in the adult male PTU treated rat. PTU treatment led to a significant reduction in GnRH-stimulated LH production. Castration resulted in 3.9- to 8.5-fold increases in circulating gonadotropin levels in both treated and control males; however, the absolute increases were significantly reduced in treated males. In contrast to circulating levels, pituitary gonadotropin contents did not increase in treated males after castration. PTU treatment did not lead to a reduction in the density of either luteotropes or folliculotropes, and both cell types increased in size and density after castration. The relative concentrations of both gonadotropin beta-subunit messenger RNAs increased more slowly in treated males than in controls after castration. Thus, although treated rats have the intrinsic ability to produce normal circulating levels of LH and FSH, gonadal feedback and an overall reduction in gonadotrope synthetic ability combine to produce the chronically reduced circulating levels of these hormones. PMID- 9202209 TI - Heat shock-induced inhibition of acute steroidogenesis in MA-10 cells is associated with inhibition of the synthesis of the steroidogenic acute regulatory protein. AB - The synthesis of heat shock proteins (HSPs) rapidly increases in cells under a broad range of stress conditions in addition to heat shock. Previous studies have shown that the induction of HSPs severely impairs the ability of steroidogenic cells to synthesize steroids in response to acute stimulation. De novo synthesis of the steroidogenic acute regulatory (StAR) protein has been shown to be indispensable for acute steroid hormone biosynthesis; however, the effect of HSP induction on the synthesis of the StAR protein has not yet been studied. In the present study we investigated whether HSP induction might influence the steroidogenic activity of MA-10 mouse Leydig tumor cells, and whether this effect may involve the synthesis of StAR protein. MA-10 cells exposed to 45 C for 10 min and allowed to recover for 2 h at 37 C displayed a 6-fold increase in HSP-70 at 3 h postrecovery and a 20-fold increase in this protein at 6 h postrecovery. This heat shock regimen also acutely inhibited both progesterone production and StAR protein synthesis in MA-10 cells in response to LH and cAMP analog stimulation. The activity and quantity of cytochrome P450 side-chain cleavage and 3beta hydroxysteroid dehydrogenase were not affected by this heat shock treatment, indicating that the loss of steroidogenic capacity was not a result of inhibition of the enzymes involved in the conversion of cholesterol to progesterone. The results suggest that the previously observed antisteroidogenic effects of heat shock treatment may be due mainly to the acute inhibition of StAR protein synthesis. PMID- 9202210 TI - Control of rat preadipocyte adipose conversion by ovarian status: regional specificity and possible involvement of the mitogen-activated protein kinase dependent and c-fos signaling pathways. AB - As ovariectomy induces obesity in rats, we have investigated the influence of ovariectomy and hormone replacement on the proliferation and differentiation capacities of rat cultured preadipocytes removed from different fat depots (femoral sc, parametrial, and perirenal). Ovariectomy induced increased proliferation and differentiation as well as high mitogen-activated protein (MAP) kinase activity and c-fos protein induction in both confluent and differentiated preadipocytes from perirenal fat depots. In parametrial preadipocytes, ovariectomy also increased proliferation and c-fos protein induction, but failed to alter the capacities of these cells to differentiate. Treatment of ovariectomized rats with estradiol and progesterone reversed the promoting effect of ovariectomy on proliferation, differentiation, and c-fos induction in perirenal preadipocytes, but not the MAP kinase activation observed during the proliferative phase. This treatment also reversed the promoting effect of ovariectomy on proliferation and c-fos induction seen in confluent parametrial preadipocytes. In contrast, sc preadipocytes were totally insensitive to ovarian status in terms of proliferation and differentiation capacities, MAP kinase activity, and c-fos induction. This study demonstrates that adipogenesis is site specifically controlled by the ovarian status in the rat. It also suggests that ovariectomy-induced obesity (mainly abdominal) could be related to changes in some of the signaling pathways controlling adipogenesis in intraabdominal preadipocytes. PMID- 9202211 TI - Neuropeptide Y Y1-receptor stimulation is required for physiological amplification of preovulatory luteinizing hormone surges. AB - Neuropeptide Y (NPY) has been shown to potentiate the actions of LHRH during the generation of preovulatory LH surges. It is not yet known, however, if activation of a specific subtype of NPY receptors in the anterior pituitary gland is an obligatory event in the stimulation of spontaneous LH surges. A battery of NPY receptor agonists, as well as the specific NPY Y1 receptor antagonist BIBP3226, were used to assess the role of Y1 receptors in the amplification of LH surges. In Exp 1, the potencies of a number of NPY agonists in facilitating LHRH-induced LH surges were assessed in pentobarbital (PB)-blocked, proestrous rats. The rank ordered potencies of these compounds were determined to be PYY = [Leu31Pro34]NPY > NPY >> hPP = rPP = NPY(13-36), which most closely reproduces the known rank ordered affinties of these compounds for the Y1 receptor. In Exp 2, a Y1 subtype- specific antagonist, BIBP3226, was administered to unanesthetized, proestrous rats to assess the involvement of the Y1 receptor in the stimulation of spontaneous LH surges. The BIBP3226 compound strongly attenuated the endogenous proestrous LH surge, reducing the integrated value of LH secretion during the proestrous surge by more than 70%. In Exp 3, we assessed the ability of the Y1 receptor antagonist to block exogenous NPY effects on LHRH-induced LH surges. Treatment with BIBP3226 was found to completely prevent NPY amplification of LHRH induced LH surges in pentobarbital-blocked, proestrous rats, thus confirming a pituitary locus of action of the drug. Taken together, these data clearly demonstrate that activation of neuropeptide Y receptors of the Y1 subtype is required for the physiological amplification of the spontaneous preovulatory LH surge in rats. PMID- 9202212 TI - Transcriptional regulation of Sertoli cell immediate early genes by interleukin-6 and interferon-gamma is mediated through phosphorylation of STAT-3 and STAT-1 proteins. AB - The immediate early genes are regulated by a variety of extracellular signals, including pleiotropic cytokines. The effects of the testicular cytokines, interleukin-6 (IL-6) and interferon-gamma (IFN-gamma), on signal transducers and activators of transcription 3 and 1 (STAT-3 and STAT-1) and on c-fos gene expression in primary Sertoli cells are suggestive of their roles in differential function. Using the tyrosine phosphorylation inhibitor, genistein, and electrophoretic mobility shift assay, we show that IL-6 and IFN-gamma induce nuclear factor STAT-3 and STAT-1 DNA-binding activity to the sis-inducible element of c-fos in a genistein-dependent pathway. Quantitative solution hybridization, Northern blot, and nuclear run-on analysis show that differential induction of c-fos, junB, and c-myc messenger RNA (mRNA) by these cytokines occur at transcriptional levels. IL-6 stimulates c-fos mRNA levels by 6-fold while increasing junB levels by 2-fold. IFN-gamma increases c-fos message 2-fold, but has no effect on junB mRNA levels. Furthermore, genistein treatment blocks the induction of c-fos and junB gene expression, demonstrating that tyrosine phosphorylation of STAT proteins is involved in the cytokine regulation of the Sertoli immediate early genes. H7, a serine/threonine phosphorylation inhibitor, also blocks c-fos gene induction by IL-6 and IFN-gamma, but does not affect the DNA-binding activities of STAT-3 and STAT-1. Finally, IL-6 treatment of Sertoli cells (3-6 h) increases the amounts of activating protein-1 binding to activating protein-1 element and c-myc transcription. PMID- 9202213 TI - Interferon-gamma-induced interferon regulatory factor-1 (IRF-1) expression in rodent and human islet cells precedes nitric oxide production. AB - The radical nitric oxide (NO) may be a mediator of beta-cell damage in IDDM. The cytokines IFN-gamma and IL-1beta are required for expression of the enzyme nitric oxide synthase (iNOS), and NO production by human pancreatic islets. In this study, possible mechanisms by which IFN-gamma participates in iNOS messenger RNA (mRNA) expression were evaluated in both rodent and human islets cells. Addition of IFN-gamma, before or after arrest of IL-1beta-induced iNOS gene transcription by actinomycin D, did not prolong iNOS mRNA half life in the rat insulin producing cell line RINm5F (RIN cells). IFN-gamma also failed to modify IL-1beta induced activation of the transcription factor kappaB (NF-kappaB) in RIN cells, as determined by electrophoretic mobility shift assay. However, IFN-gamma induced an early (30 min(-1) h) increase in interferon regulatory factor-1 (IRF-1) mRNA expression and a later (2 h) 19-fold increase in RIN cell nuclear IRF-1 protein content, an effect further potentiated by IL-1beta. The total cellular content of IRF-1 protein increased by 30- to 50-fold in human islets exposed for 2-8h to IFN gamma or IFN-gamma + IL-1beta. IL-1beta alone induced a marginal and transient increase in IRF-1. It has been previously reported that nicotinamide prevents IL 1beta-induced IRF-1 expression in rat pancreatic islets. However, nicotinamide (20 mM) presently failed to prevent IL-1beta + IFN-gamma-induced IRF-1 protein expression in human pancreatic islets. In conclusion, the effects of IFN-gamma on iNOS expression can neither be explained by iNOS mRNA stabilization nor increased NF-kappaB activation. However, IFN-gamma induces an early increase in cellular IRF-1 content, and this may contribute to increased iNOS mRNA expression. PMID- 9202214 TI - Characterization of multiple promoters directing tissue-specific expression of the human gonadotropin-releasing hormone gene. AB - Two promoters directing tissue-specific expression of GnRH gene in neuronal and reproductive tissues were characterized by functional analyses of GnRH promoter luciferase (LUC) constructs in transfected placental cells (JEG) and hypothalamic neuronal cells (GT1-7). Results indicate that the downstream promoter directs the expression in a neuronal cell-specific manner, whereas the upstream promoter is fully active in the nonneural placental cell line. Transfection studies carried out in several tumor cell lines derived from human reproductive tissues verified that the upstream GnRH promoter construct was much more active in directing luciferase expression in reproductive tissue. The use of both upstream and downstream promoters in various human tumor cell lines derived from reproductive tissues were demonstrated by RT-PCR. Our studies also demonstrate that the reproductive tissue-specific messenger RNA transcribed from upstream promoter is capable of directing synthesis of preproGnRH protein. Serial deletion studies localized a cell-specific upstream promoter region that directs reproductive tissue expression. DNase I footprint analysis using nuclear extract obtained from the JEG cells indicated DNA/protein interactions in four specific sequence elements of the upstream promoter region. The interaction between nuclear binding proteins present in the JEG cells (but not the GT1-7 cells) and the four specific sequences in the upstream promoter region was confirmed by gel mobility shift analysis. PMID- 9202215 TI - Effects of RU486 on estrogen, progesterone, oxytocin, and their receptors in the rat uterus during late gestation. AB - Oxytocin (OT) and its receptor (OTR) are synthesized in the endometrium and myometrium of the pregnant rat during late gestation. Both are regulated by estrogen and progesterone (P4), and tissue concentrations of both increase markedly before parturition. The P4 antagonist RU486 will induce parturition in the rat. The purpose of the present studies was to investigate changes in OT and OTR messenger RNA (mRNA) and peptide synthesis within the pregnant rat uterus during RU486-induced parturition. Pregnant rats were given a single injection of RU486 (2.5 mg/rat in oil) on day 15 of pregnancy (normal delivery occurs on day 22). Control animals received injections of oil only. Groups of animals (n = 5 in each group) were euthanized at 0, 6, 12, 24, and 48 h after injection and during labor (immediately after delivery of the first pup). Maternal serum estradiol (E2), P4 and uterine OT, and PGE2 concentrations were measured by RIA. Prostaglandin F2alpha and estrogen receptor levels were measured by enzyme immunoassay (EIA). OTR and P4 receptor (PR) were measured using radioligand binding assays. OT, OTR, and estrogen receptor mRNAs were measured with ribonuclease protection assays. The average time to delivery, after RU486 injection, was 27.0 +/- 1.2 h. Serum E2 and P4 levels were increased slightly, but significantly, at 24 h after RU486. In controls, OT mRNA increased significantly, and this increase was blocked in the RU486 treatment group. OTR mRNA levels increased within 6 h of RU486 and remained elevated until delivery. OTR peptide was increased by 12 h. PGE2 and PGF2alpha were increased 3-fold and 16-fold, respectively, but not until after the increase in OTR had occurred. We conclude that the mechanism of action of RU486 is to inhibit the P4 suppression of OTR synthesis, allowing increased expression of OTR, which may directly stimulate myometrial contractions or act indirectly through increased synthesis of PGs. PMID- 9202216 TI - Impaired cytosolic Ca2+ response to glucose and gastric inhibitory polypeptide in pancreatic beta-cells from triphenyltin-induced diabetic hamster. AB - Oral administration of a single dose of triphenyltin compounds induces diabetes with decreased insulin secretion in rabbits and hamsters after 2-3 days without any morphological changes in pancreatic islets. In the present study, to test the possibility that the impaired insulin secretion induced by triphenyltin compounds could result from an impaired Ca2+ response in pancreatic beta-cells, we investigated the effect of triphenyltin-chloride (TPTCl) administration on the changes in the cytoplasmic Ca2+ concentration ([Ca2+]i) induced by secretagogues, such as glucose, high K+, gastric inhibitory polypeptide (GIP), and acetylcholine (ACh) in hamster pancreatic beta-cells. TPTCl administration caused partial suppression in 10 mM K+-induced rise in [Ca2+]i without suppressing the increase in [Ca2+]i evoked by 20-50 mM K+. Administration of TPTCl strongly inhibited the rises in [Ca2+]i induced by 27.8 mM glucose, 100 microM ACh in the presence of 5.5 mM glucose, and by 100 nM GIP in the presence of 5.5 mM glucose. In the ACh induced response, TPTCl administration strongly suppressed the late sustained phase, while weakly suppressing the initial rise in [Ca2+]i. TPTCl administration significantly suppressed the rise of cAMP content in islet cells induced by 100 nM GIP with 1 mM 3-isobutyl-1-methylxanthine in the presence of 5.5 mM glucose (P < 0.01, N = 5-11). TPTCl administration also impaired the insulin secretion in islet cells induced by 27.8 mM glucose, 100 nM GIP in the presence of 5.5 mM glucose, and 100 microM ACh in the presence of 5.5 mM glucose (P < 0.05, N = 9 16). We conclude that the pathology of triphenyltin-induced diabetes in hamsters involves a defect in cellular Ca2+ response due to a reduced Ca2+-influx through voltage-gated Ca2+ channels. PMID- 9202218 TI - Orthovanadate stimulates cyclic guanosine monophosphate-inhibited cyclic adenosine monophosphate phosphodiesterase activity in isolated rat fat pads through activation of particulate myelin basic protein kinase by protein tyrosine kinase. AB - Involvement of protein kinases in the stimulation of cGMP-inhibited cAMP phosphodiesterase (PDE) activity by orthovanadate (vanadate) was studied. When the fat pads were incubated with 2 mM vanadate or 10 nM insulin, the stimulation of myelin basic protein kinase (MBPK) activity in the particulate by vanadate reached a maximum at 60 min. In contrast, insulin showed a transient increase at 20 min. A 60-min incubation of the fat pads with vanadate stimulated all activities of protein tyrosine kinase (PTK), MBPK, and PDE in the particulate, in a similar dose-dependent manner. Amiloride, a PTK inhibitor, inhibited the stimulations of three enzymes by vanadate in a similar concentration range. Enzyme fractions, which were separated from the solubilized particulate, were subjected to the immunoblot analysis. A fraction of MBPK was identified to contain a major protein of mol wt (44K) and a minor one (42K), both of which are immunoreactive with a mitogen-activated protein kinase (MAPK) antibody. The partially purified PDE activity was stimulated by the addition of the partially purified MBPK. The further stimulation was observed with the PTK-activated MBPK. These results suggest that vanadate stimulates in part the PDE activity through the activation of the particulate MBPK, probably MAPKs, by PTK sensitive to vanadate. PMID- 9202219 TI - Expression of urokinase-type plasminogen activator and its receptor during ovarian follicular development. AB - Although tissue-type plasminogen activator (tPA) and plasminogen activator inhibitor-1 (PAI-1) are believed to be involved in the biochemical cascade leading to extracellular matrix degradation during ovulation, the presence and possible role of urokinase-type PA (uPA) and its receptor (uPAR) in follicular wall remodeling during follicular development are poorly understood. In the current studies, we have examined their presence in the rat ovary and compared the changes in both uPA and uPAR expression with those of tPA and PAI-1 during follicular growth in vivo. The presence of these proteins in various follicular cells at different stages of maturation was evaluated by immunolocalization and ELISA. Abundance of respective messenger RNA in granulosa cells from preantrallearly antral, midantral and preovulatory follicles and the residual ovaries was determined by Northern blot analysis. Whereas uPA transcript and protein levels were highest at the earliest stage of follicular growth examined and decreased markedly before the expected time of ovulation, the opposite was true for uPAR. In addition, tPA and PAI-1 messenger RNA abundance and protein contents were low in both granulosa and residual ovarian tissue during early follicular development but increased thereafter, reaching highest levels at the preovulatory period. These findings demonstrate for the first time the presence of uPAR in ovarian follicles and its developmental expression. The coincidental rise in uPAR and PAI-1 proteins during the preovulatory period may be important for the regulation of extracellular matrix remodelling before ovulation. The reciprocal expression of uPA and tPA during follicular development are consistent with the notion that these proteases have different biological functions in the ovary, i.e. tPA is involved in follicular wall remodelling before ovulation whereas uPA is important in extracellular matrix degradation during cell proliferation and migration that accompany follicle growth. PMID- 9202217 TI - TNF alpha-mediated inhibition and reversal of adipocyte differentiation is accompanied by suppressed expression of PPARgamma without effects on Pref-1 expression. AB - Tumor necrosis factor alpha (TNF alpha) is a polypeptide hormone with pleiotropic effects on cellular proliferation and differentiation. To investigate how TNF alpha inhibits and reverses adipocyte differentiation, we studied the expression of two factors involved in the adipocyte differentiation process. Peroxisome proliferator-activated receptor gamma (PPARgamma) is a positive regulator of adipogenesis, whereas preadipocyte factor 1 (Pref-1) inhibits adipocyte differentiation. The expression patterns of both PPARgamma and Pref-1 change during early stages of adipocyte differentiation. Decreased expression of Pref-1 and increased expression of PPARgamma occur 1 day and 2 days, respectively, after 3T3-L1 cells reach confluence. During TNF alpha-mediated inhibition of adipocyte differentiation, PPARgamma messenger RNA (mRNA) expression stays at low levels. In contrast, TNF alpha treatment has no effect on the normal decrease in Pref-1 gene expression that occurs during adipogenesis. We observed that certain cytokine and growth factors [such as TNF alpha, basic fibroblast growth factor, transforming growth factor beta, and protein kinase C-activating agents plus calcium ionophore], when added to differentiated adipocytes, cause rapid down regulation of PPARgamma mRNA expression with concomitant decrease in adipocyte specific gene expression but fail to increase Pref-1 mRNA expression. Moreover, addition of TNF alpha to fully differentiated adipocytes results in the rapid disappearance of PPARgamma protein expression and the rapid loss of PPARgamma DNA binding activity. Therefore, Pref-1 seems to function as a nonreversible molecular checkpoint whose expression is insensitive to TNF alpha-generated signals, whereas PPARgamma expression remains sensitive to TNF alpha at all stages of the adipogenesis program. Our results support the notion that dedifferentiated adipocytes and preadipocytes are not identical, though they share many similar morphological and gene expression patterns. PMID- 9202220 TI - An intramolecular disulfide bond between conserved extracellular cysteines in the gonadotropin-releasing hormone receptor is essential for binding and activation. AB - In this study, site-directed mutagenesis and biochemical strategies have been used to establish whether disulfide bonding between extracellular Cys residues contributes to the structural integrity of the GnRH receptor (GnRH-R) and, if so, to delineate the nature of the bonding patterns involved. The majority of G protein-coupled receptors (GPCRs) contain a pair of conserved Cys residues in the first and second extracellular domains, and these residues have been shown to form a receptor stabilizing disulfide bridge structure. However, many GPCRs contain other nonconserved Cys residues, and in some GPCRs these have also been shown to contribute to receptor integrity and stability. The rat GnRH-R contains four extracellular Cys residues. Two are conserved throughout the GPCR superfamily and lie at positions Cys114 and Cys195 in the first and second extracellular loops, respectively. The other two Cys residues occupy nonconserved positions at Cys14 in the amino terminus and Cys199 in the second extracellular loop. To assess the role of extracellular Cys residues in disulfide bonding interactions, each of these residues were mutated to Ala, expressed in COS-1 cells, and ligand binding and second messenger properties ascertained. To monitor levels of wild-type (WT) and mutant receptor cell surface expression, a hemagglutinin (HA) epitope tag was incorporated into the receptor constructs (GnRH-R WT, Cys14Ala, Cys114Ala, Cys195Ala, and Cys199Ala). Cys199Ala mutant maintained levels of receptor binding and second messenger production comparable with the WT GnRH-R, whereas mutant Cys14Ala exhibited some ligand binding and functional receptor activity, albeit at a reduced level. Mutations Cys114Ala and Cys195Ala showed no functional responses despite displaying levels of cell surface expression similar to the WT receptor. Specific binding of the WT and mutant receptors Cys14Ala and Cys199Ala was inhibited in the presence of the disulfide bond reducing agent, DTT, implying that disulfide bonds are formed and can be reduced in these mutant receptors. This study demonstrates that GnRH-R residues Cys114 and Cys195 have a disulfide bonding interaction role essential for the maintenance of receptor function. In contrast, Cys14 and Cys199 are not involved in disulfide bonding that is required for ligand binding or second messenger production. PMID- 9202221 TI - Thyrotropin-releasing hormone gene expression by anterior pituitary cells in long term cultures is influenced by the culture conditions and cell-to-cell interactions. AB - It has been suggested that TRH, synthesized by anterior pituitary (AP) cells in long-term monolayer cultures, may act as a paracrine or autocrine regulator. Because local control through messenger molecules depends on the cellular microenvironment, we were interested in studying the synthesis of TRH by AP cells in different culture systems and under various conditions. When AP cells were cultured as monolayers in medium containing 10% FCS for long periods of time (up to 3 weeks), a considerable increase in TRH content and prepro-TRHmessengerRNA (preproTRHmRNA) levels could be demonstrated by RIA and Northern blot analysis, whereas the cellular content of the TRH-like peptide pyroGlu-Glu-Pro-NH2 decreased with time in culture to undetectable levels. The release of TRH could be stimulated by depolarizing concentrations of K+ (55 mM), by the Ca++ ionophore A23187, and by GnRH, but not by CRH or GRF, indicating that TRH is stored in gonadotropes. Moreover, a combined in situ hybridization and immunocytochemical analysis demonstrated colocalization of LH in preproTRHmRNA-positive AP cells. When AP cells were cultured as reaggregates in the same (FCS-containing) medium, only a marginal increase in TRH content and preproTRHmRNA levels was observed. Irrespective of the culture systems and the culture conditions used, TRH gene expression was not observed when FCS was omitted. These results indicate that TRH gene expression more likely reflects derepression, rather than induction, of the TRH gene. PMID- 9202222 TI - Tissue-specific expression of the bovine aromatase-encoding gene uses multiple transcriptional start sites and alternative first exons. AB - Here we report on the genomic structure of the bovine aromatase cytochrome P450 encoding gene (Cyp19) and its tissue-specific transcript variants. The gene comprises at least 14 exons (1.1, 1.2a, 1.2b, 1.3,1.4, and 2-10) spanning more than 56 kilobases of genomic DNA. The coding area is confined to exons 2-10. Transcriptional start sites of Cyp19 were examined in granulosa cells, placenta, testis, adrenal gland, and brain, employing 5'-RACE (rapid amplification of complementary DNA ends) and primer extension. The analysis of 5'-RACE clones revealed six Cyp19 transcript variants that were different within their 5' untranslated regions (5'-UTR). Yet, the coding region was identical in all clones. Although two of these 5'-UTR (the first 152 nucleotides of exon 2 and exon 1.4) are conserved among different species, four others (exons 1.1, 1.2a, 1.2b, and 1.3) did not show sequence homology to any other species. Transcription from exons 1.1 and 2 starts at several adjacent sites. In granulosa cells and placenta, but not in brain, a fraction of transcripts starting with exon 1.2a contains an additional untranslated exon, 1.2b, due to alternative splicing. Transcript variants comprising exon 1.1, 1.2a, 1.2b, or 1.3 were mainly found in the placenta, those with the 5'-UTR of exon 2 were predominant in granulosa cells, and transcripts with exon 1.4 prevailed in the brain. Estimates of Cyp19 transcript concentrations in six different tissues revealed high levels in granulosa cells and placenta, intermediate levels in testis and brain, and low levels in adrenal gland and liver. Our experiments demonstrate that six transcript variants of the bovine Cyp19 gene, including 9-11 exons, are expressed with tissue-specific preferences. These transcripts are presumably generated using five different promoter regions and tissue-specific alternative splicing. PMID- 9202223 TI - Glucocorticoid-induced differentiation of fetal rat calvarial osteoblasts is mediated by bone morphogenetic protein-6. AB - Glucocorticoids (GCs) at physiological concentrations promote osteoblast differentiation from fetal calvarial cells, calvarial organ cultures, and bone marrow stromal cells; however, the cellular pathways involved are not known. Bone morphogenetic proteins (BMPs) are recognized as important mediators of osteoblast differentiation. Specific roles for individual BMPs during postembryonic membranous bone formation have yet to be determined. We recently reported that GC potentiated the osteoblast differentiation effects of BMP-2 and BMP-4, but not of BMP-6, which, by itself, was the most potent of the three. In the present study, we used fetal rat secondary calvarial cultures to study the role of BMP-6 during early osteoblast differentiation. Treatment with the GC triamcinolone (10(-9) M) resulted in a 5- to 8-fold increase in BMP-6 steady-state messenger RNA levels, peaking at 12 h. In contrast, BMPs -2, -4, -5, -7, and transforming growth factor (TGF)-beta1 messenger RNA levels increased by less than 2-fold, after GC treatment, compared with untreated control cultures at 24 h. BMP-6 protein secretion increased 6- to 7-fold by 12 h and 12-fold (from 7.5 to 90 ng/ml) by 24 h, as measured by quantitative Western analysis. Treatment of cells with oligodeoxynucleotides antisense to BMP-6 diminished secretion of BMP-6 protein and significantly inhibited the GC-induced differentiation, as determined by a 10 fold decrease in the number of mineralized bone nodules, compared with controls that were treated with sense oligonucleotides or no oligonucleotides (ANOVA, P < 0.05). The antisense oligonucleotide inhibition of differentiation was rescued by treatment with exogenous recombinant human BMP-6. We conclude that GC-induced differentiation of osteoblasts from the pluripotent precursors is mediated, in part, by BMP-6. These results suggest that BMP-6 has an important and unique role during early osteoblast differentiation. PMID- 9202224 TI - Central oxytocin administration reduces stress-induced corticosterone release and anxiety behavior in rats. AB - Endocrine responses to noise stress and anxiety-related behaviors were measured in groups of ovariectomized, estradiol-treated female rats given central infusions of oxytocin. Control animals receiving isotonic saline showed a large increase in plasma corticosterone concentrations in response to 10 min of white noise. This response to noise stress was significantly and dose dependently decreased by oxytocin administered intracerebroventricularly at 10 or 100 ng/h for 5 days. Oxytocin also significantly decreased rearing behavior during this stress. When a second noise stress was given 3 days after cessation of oxytocin infusion, corticosterone responses did not differ between the control and previously oxytocin-infused animals. Administration of vasopressin had no significant effect on either the corticosterone or behavioral responses to noise stress. Anxiety-related behaviors were measured on the elevated plus-maze. No significant differences were seen in maze exploration between saline- and oxytocin-treated animals when housed and tested in the same environment. However, when animals were mildly stressed by testing in an unfamiliar environment, oxytocin-treated animals showed a higher proportion of open arm entries and spent significantly more time in the open arms of the maze. Thus, oxytocin exerts a central anxiolytic-like effect on both endocrine and behavioral systems and could play a role in moderating behavioral and physiological responses to stress. PMID- 9202225 TI - Proliferin induces endothelial cell chemotaxis through a G protein-coupled, mitogen-activated protein kinase-dependent pathway. AB - To investigate the mechanism of action of the placental angiogenic hormone proliferin (PLF), we analyzed the signaling components in endothelial cells that are required for PLF-induced chemotaxis. Pertussis toxin, which inactivates Gi proteins, inhibited PLF-induced chemotaxis of endothelial cells. Gi proteins can lead to activation of the mitogen-activated protein kinase (MAPK) pathway; PLF was found to stimulate MAPK activity, and this induction was blocked by both pertussis toxin and a specific inhibitor of MAPK kinase, PD 098059. Furthermore, a blockade of MAPK activation prevented endothelial cell movement in response to PLF. As PLF functionally interacts with the insulin-like growth factor II (IGF II)/mannose 6-phosphate receptor, we also examined the effects of pertussis toxin and PD 098059 on another ligand for this receptor, a mutant form of IGF-II; both inhibitors also block the action of this factor on endothelial cells. These data suggest that chemotaxis initiated by PLF and mediated by the IGF-II/mannose 6 phosphate receptor occurs through a G protein-coupled pathway, and that MAPK activation is necessary for the chemotactic response. PMID- 9202226 TI - Pituitary follistatin regulates activin-mediated production of follicle stimulating hormone during the rat estrous cycle. AB - Follistatin, an activin-binding protein, plays a key role in the modulation of activin-dependent functions. In the anterior pituitary, activin stimulates the synthesis and secretion of FSH. In the current study, we assessed the roles of locally produced activin and follistatin in the control of FSH gene expression and secretion. The anterior pituitary gland follistatin content was measured at frequent intervals during the rat estrous cycle. Follistatin protein levels were high before the primary gonadotropin surges, decreased by 50% on proestrous evening, and rebounded to a peak at midnight on proestrus before returning to presurge levels on estrus morning. Changes in pituitary follistatin protein content were preceded by parallel changes in pituitary follistatin messenger RNA (mRNA). The trough in follistatin protein content on proestrus coincided with a peak in circulating levels of free activin A (not bound to follistatin) and a sharp rise in FSHbeta mRNA levels, suggesting that decreased pituitary follistatin leads to increased available activin. To quantitate the contribution of pituitary free activin to pituitary expression of FSHbeta mRNA and the primary and secondary serum FSH surges, rats were infused through carotid catheters with saline or recombinant human follistatin (288-amino acid isoform; rhFS-288) at different times during the estrous cycle. Infusion of rhFS-288 on diestrus did not affect FSH production. In contrast, infusion of rhFS-288 during the secondary FSH surge decreased the peaks in FSHbeta mRNA and serum FSH by 63% and 47%, respectively, relative to those in saline-infused control animals. Infusion of rhFS-288 during the primary FSH surge decreased serum FSH to a lesser degree (24%). These data indicate a physiological role for pituitary follistatin in the control of activin-mediated FSH synthesis and secretion during the rat estrous cycle. PMID- 9202227 TI - Stimulatory effect of human, but not bovine, growth hormone expression on numbers of tuberoinfundibular dopaminergic neurons in transgenic mice. AB - Mice transgenic for heterologous and ectopic GH expression serve as models for studying the feedback effects of elevated nonregulated GH on hypothalamic hypophysiotropic neurons as well as on peripheral function. For example, hypothalamic somatostatin expression has been shown to be increased markedly in mice bearing either bovine (b) or human (h) GH transgenes. Human, but not bovine, GH has lactogenic properties in mice, and appears to stimulate PRL-inhibiting tuberoinfundibular dopaminergic (TIDA) neurons. The present study was designed to determine the effect of a lifelong excess of hGH on dopamine (DA) expression in and numbers of TIDA neurons. Male mice of four transgenic lines were examined. The transgenic animals bore constructs of either bGH or hGH fused to either metallothionein (MT) or phosphoenolpyruvate carboxykinase (PEPCK) promoters; brains of transgenic mice were compared morphologically with those of nontransgenic littermates. Formaldehyde-induced catecholamine histofluorescence and tyrosine hydroxylase (TH) immunocytochemistry were examined in alternate brain sections; cell number was quantified for TIDA neurons (area A12) and a nonhypophysiotropic diencephalic DA area, the medial zona incerta (A13). Body weights were higher (P < 0.01) in PEPCK-GH than in MT-GH transgenic mice, as were serum levels of heterologous GH in those lines. In MT-hGH, but not MT-bGH or PEPCK-bGH, transgenic mice, A12 perikaryal fluorescence was enhanced, and ME fluorescence was reduced compared with those in control animals. The reduced ME DA is likely to reflect stimulation of TIDA neurons, because A12 TH immunoreactive neuron number was increased by 34% in MT-hGH mice compared with that in controls (P < 0.05). In mice bearing the PEPCK-hGH construct, A12 TH neuron number was increased 47% (P < 0.001) compared with that in littermate controls. There were no differences in A13 cell number among animals, and A12 cell numbers in mice expressing bGH did not differ from control values. These results suggest that although extremely high levels of circulating bGH do not stimulate TIDA neurons, lifelong high levels of hGH have a stimulatory and graded effect on developmental differentiation of these cells for TH and DA production, supporting the concept of PRL as a trophic factor for TIDA neurons. PMID- 9202228 TI - Bovine insulin-like growth factor binding protein-3: organization of the chromosomal gene and functional analysis of its promoter. AB - Insulin-like growth factor binding protein-3 (IGFBP-3), the major IGFBP in the circulation, is synthesized by the vascular endothelium in vivo and has been shown to be an important modulator of the physiological effects of IGF. IGFBP-3 is regulated by a number of growth factors/cytokines to which the vascular endothelium is exposed, including IGF-I stimulation and TGF-beta1 inhibition of IGFBP-3 in cultured endothelial cells. To understand the mechanisms of transcriptional regulation of IGFBP-3, we have cloned the bovine IGFBP-3 gene and begun the functional analysis of its promoter. Southern analysis indicated a single copy gene. The gene spanned approximately 10 kb and was divided into five exons, the fifth containing the 3' untranslated region. The transcription start site was 137 bp upstream of the initiation codon and a TATA box was located 26 bp 5' to this CAP site. No CAAT box was present but a GC rich sequence element, containing two overlapping putative AP-2 binding elements, was located 5' to the TATA box. Transient transfection studies with a series of 5' truncated luciferase reporter constructs were conducted in primary cultures of bovine aorta endothelial cells. Results of the transfection studies indicated that 1) nearly 80% of the maximal basal promoter activity was retained within the first 130 bp of the 5' flanking sequence; 2) this region responded to IGF-I, despite lacking the TTF-1/TTF-2 (thyroid specific transcription factors) binding elements that are required for IGF-I stimulation of thyroglobulin synthesis. These binding elements have also been suggested to be involved in IGF-I regulation of IGFBP-3 transcription, thus, implying the existence of novel cis-acting elements that mediate the IGF-I stimulation of bovine endothelial cell IGFBP-3 mRNA synthesis; 3) deletion of the GC rich sequence element resulted in a 60% reduction in basal promoter activity as well as loss of the IGF-1 stimulatory effect; 4) the TGF beta1 mediated inhibition of IGFBP-3 transcription required sequence element(s) beyond 1.5 kb of its promoter. PMID- 9202229 TI - Vectorial production of interleukin 1 and interleukin 6 by rat Sertoli cells cultured in a dual culture compartment system. AB - The bidirectional production of interleukin-1 (IL-1) and IL-6 by Sertoli cells and its regulation by inflammatory and physiological stimuli has been studied using a dual compartment culture system allowing the study of Sertoli cell apical and basal secretory activities. Another Sertoli cell activity, the vectorial transferrin production was also studied in all culture conditions. A low constitutive IL-1 production appeared equally distributed between both poles, while IL-6 and transferrin constitutive production was predominantly directed apically. Two activators of macrophages, lipopolysaccharides and zymosan, were found to induce marked increases of IL-1 in the compartment where they had been added: basal if added to the lower compartment and vice versa. In contrast, after a basal stimulation, IL-6 production was mainly increased in the upper compartment that corresponds to a Sertoli cell apical flux. In this system, IL-1 and IL-6 levels were not modified by FSH; they were not also affected by residual bodies and latex beads, probably due to the fact that, in the bicameral system, phagocytosis is restricted to the Sertoli cells situated at the surface of the inner compartment. IL-1beta, but not IL-1alpha, induced IL-6 secretion in the compartment of stimulation. In conclusion, the present study demonstrates that vectorial secretory patterns of IL-1 and IL-6 production greatly differ and that these cytokines are also differently regulated. These results suggest that Sertoli IL-1 and IL-6 have different targets within the testis and that, in normal and pathophysiological conditions, both the tubular and the interstitial compartments may be influenced by the action of these paracrine factors. PMID- 9202230 TI - Glutathione peroxidase degrades intracellular hydrogen peroxide and thereby inhibits intracellular protein iodination in thyroid epithelium. AB - Protein iodination in the thyroid is largely confined to the surface of the epithelium. Intracellular iodine binding is insignificant. We have tested our hypothesis that the key mechanism in the control of intracellular iodination is the control of the intracellular availability of H2O2. The sites of iodination were identified by locating bound radioiodine in electron microscopic autoradiographs, produced from porcine thyroid epithelium grown on filter in Transwell bicameral culture chambers. Autoradiographs obtained after standard incubations with 125I for 15 min to 3 h were all characterized by concentrations of autoradiographic grains along the external surface of the plasma membrane and very few grains over the cytoplasm. The presence of 10 microM H2O2 in the incubation medium resulted in a drastically changed labeling pattern now showing a dissemination of grains over the entire cytoplasm. Epithelia with elevated GSH peroxidase activity produced autoradiographs showing the same restriction of grains to the cell surface as controls; this pattern was the same in the absence and presence of H2O2 (up to 10 microM). Cultures with subnormal GSH peroxidase activity presented cytoplasmic labeling both in the absence and presence of H2O2. In conclusion, iodine binding in filter-cultured thyroid epithelium under normal conditions is an extracellular process located at the cell surface. When H2O2 is available intracellularly, iodination takes place in the cytoplasm, evidently catalyzed by intracellular thyroperoxidase. Normally, this iodination is prevented by cytosolic GSH peroxidase that effectively degrades H2O2 and thus controls intracellular iodination. The observations should be applicable to the thyroid in vivo. PMID- 9202231 TI - Hyperinsulinemia-induced hypoglycemia is enhanced by overexpression of connexin 43. AB - To assess whether cell to cell communications via connexins (Cx) participate to insulin secretion in vivo, we studied insulinoma cells (INS1) implanted in rats after stable transfection with connexin 43 (Cx43). We found that compared to wild type and transfected cells, which in vivo express modest levels of Cx43 and junctional communication, cells overexpressing Cx43 communicated extensively, featured decreased growth, and induced a much higher hyperinsulinemia. As a result, rats with insulinomas made of these cells became more severely hypoglycemic than rats implanted with either wild-type, neomycin-transfected cells or cells transfected with a Cx43 antisense complementary DNA. Rats implanted with transfected cells that expressed modest level of Cx43 showed levels of circulating insulin similar to those in rats implanted with wild-type INS1 cells. The data show that overexpression of Cx43 influences the growth and secretion of the implanted insulinoma cells, providing evidence for a contribution of Cx-mediated cell to cell communication in the functioning of insulin-producing cells in vivo. PMID- 9202232 TI - Expression of 17beta-hydroxysteroid dehydrogenase type 1 and type 2, P450 aromatase, and 20alpha-hydroxysteroid dehydrogenase enzymes in immature, mature, and pregnant rats. AB - In the present study, we evaluated the expression and regulation of 17beta hydroxysteroid dehydrogenase (17HSD) type 1 and type 2, cytochrome P450 aromatase (P450arom), and 20alpha-hydroxysteroid dehydrogenase (20HSD) in mature and pregnant rats. Immunohistochemical analysis of rat 17HSD type 1 showed that the enzyme is exclusively expressed in the granulosa cells of developing, healthy, primary, secondary, and tertiary follicles at all stages of the estrous cycle and pregnancy, and is not detected in the corpora lutea. The data showed that the amount of the enzyme expressed in the follicle increases as follicular maturation progresses and is highest in tertiary and Graafian follicles. However, Northern blot analysis of total RNA from whole ovaries showed a rather constitutive expression of the 17HSD type 1 enzyme. It is evident that compared with P450arom, 17HSD type 1 is more widely expressed in the follicles during the various maturational stages of folliculogenesis. Hence, the data indicate distinct localization, expression, and regulation patterns for 17HSD type 1 and P450arom during the rat estrous cycle and pregnancy. Furthermore, compared with the two estradiol biosynthetic enzymes, a different expression pattern was detected for 20HSD messenger RNA. During the estrous cycle the enzyme was detected in the ovaries throughout the cycle, and in the ovaries of pregnant animals the enzyme showed an expression pattern the opposite of that observed for P450arom. Rat 17HSD type 2, not detected in the ovaries, was constitutively expressed in both female and male liver and small intestine in 21-day-old fetuses up to 6-week-old mature animals. Similarly, in these tissues the enzyme was constitutively expressed in normal cycling and pregnant animals, but it showed increasing expression in the placenta as pregnancy advanced. The relatively constitutive expression of the enzyme at all physiological stages of the animals suggests a general role for the enzyme in the inactivation of circulating sex steroids. PMID- 9202233 TI - Evidence that the thyrotropin receptor ectodomain contains not one, but two, cleavage sites. AB - TSH receptor (TSHR) cleavage into two subunits (A and B) was explored using two new mammalian cell lines expressing the recombinant receptor; 1) TSHR-10,000 CHO cells overexpressing the TSHR; 2) TSHRmyc cells with a c-myc epitope inserted at residues 338-349. Immunoprecipitation or immunoblotting of TSHR-10,000 cells with mAb to either the A subunit or the B subunit revealed multiple forms of the TSHR: 1) uncleaved receptors of approximately 115 kDa and approximately 100 kDa with complex carbohydrate and high mannose carbohydrate, respectively; 2) two subunit TSHR with an approximately 62 kDa A subunit containing complex carbohydrate. The A subunit was approximately 35 kDa after enzymatic deglycosylation (predicted C terminus near residue 330). The nonglycosylated B subunit was evident primarily as an approximately 42 kDa band (predicted N terminus near residue 380). The sum of the A and B subunit polypeptide backbones was smaller than the predicted size of the TSHR, a polypeptide backbone (84.5 kDa), raising the possibility that an approximately 5-kDa polypeptide fragment was excised during intramolecular cleavage. This hypothesis was supported by data obtained with the TSHRmyc cells. Thus, mAb to the c-myc epitope and to amino acid residues 22-35 (mAb A10) were equally effective in detecting the single chain forms of the TSHR in these cells. However, the 35 kDa, deglycosylated A subunit was clearly visible on immunoprecipitation with mAb A10 to the TSHR amino terminus, but not with the anti-myc mAb, indicating loss of the c-myc epitope at residues 338-349. Further, even though the A subunit was not detected in TSHRmyc cells with anti-myc mAb, 125I-TSH cross-linking to the cell surface showed similar A subunit expression in TSHRmyc and wild-type TSHR expressing cells. In summary, our study provides a surprising and novel finding for G protein-coupled receptors. Contrary to the prevailing concept of one cleavage site in the TSHR, we present evidence that there are, in fact, two such sites. The TSHR, like insulin, may release a C peptide during intramolecular cleavage into two subunits. PMID- 9202234 TI - Transcriptional activities of estrogen and glucocorticoid receptors are functionally integrated at the AP-1 response element. AB - Estrogens and glucocorticoids often act in opposition to regulate physiological responses. We investigated whether this might reflect the opposing actions of hormone-bound receptors on target genes regulated by the AP-1 response element. We performed a series of transfection experiments in which transcriptional activation, mediated by the AP-1 response element, was reflected in reporter gene activity. As previously described, we found that estrogens stimulate, whereas the glucocorticoid dexamethasone (Dex) inhibits, transcription through a model promoter from the collagenase gene (-73 to +63). This promoter bears a consensus AP-1 response element. When HeLa cells were treated with both estradiol and Dex, the steroids counteracted each other's transcriptional effects. The amount of transfected estrogen and glucocorticoid receptors (ER and GR) determined the extent to which Dex blunted estrogen stimulation or estrogen prevented Dex inhibition. The ER/GR interaction was observed both in the presence of estradiol and tamoxifen, which has previously been shown to have estrogen-like action at an AP-1 response element. The AP-1 family member c-Jun enhanced Dex inhibition and estradiol stimulation of transcriptional activation. c-Fos potentiated the effect of cotransfected c-Jun on estradiol stimulation but not Dex inhibition. The pattern of steroid responses was retained in the presence of the c-Jun activator phorbol 12-myristate 13-acetate. However, estradiol stimulation was lost in the presence of the c-Jun activator tumor necrosis factor-alpha. The ER/GR/AP-1 response element interaction was present, not only in a cell line originally derived from a uterine cervical adenocarcinoma (HeLa), but also in a cell line derived from the hypothalamus (GT1-1). Lastly, both progesterone receptor types A and B also interacted with the ER at the AP-1 site. These data indicate that opposing steroid influences can be mediated at the level of transcription through the AP-1 site and suggest that the integration of hormone action at this response element may underlie some of the opposing actions of estrogens and glucocorticoids or progestins on physiological responses. PMID- 9202235 TI - An antisense oligodeoxynucleotide to lipocortin 1 reverses the inhibitory actions of dexamethasone on the release of adrenocorticotropin from rat pituitary tissue in vitro. AB - Our previous studies have demonstrated that lipocortin 1 (LC1, also called annexin 1) is an important mediator of glucocorticoid action in the neuroendocrine system, particularly with regard to the powerful inhibitory actions of the steroids on the secretion of ACTH and its hypothalamic releasing hormones. In the present study, we have used an antisense oligodeoxynucleotide (ODN) unique to LC1 to investigate further the role of this protein in the regulatory effects of dexamethasone on ACTH release in vitro from rat anterior pituitary cells. Pituitary cells dispersed with collagenase retained their functional and morphological integrity in vitro and sequestered ODNs in a time dependent manner from the incubation medium. LC1 was readily detected in the cells by Western blot analysis or by immunoprecipitation/autoradiography after preloading with 35S-methionine/cysteine; the bulk of the protein was contained within an intracellular pool but a small amount was attached to the outer cell surface (pericellular). Dexamethasone (100 nm, 2.5 h) initiated de novo synthesis of LC1; it also increased the amount of LC1 in the pericellular pool detected by either method and caused a concomitant decrease in intracellular LC1. The responses to the steroid were prevented by the inclusion in the medium of an LC1 antisense ODN (50 nM, 3.5 h) but the corresponding sense and scrambled ODN sequences were inert. None of the ODN sequences tested influence the expression of annexin 5 in the pituitary tissue. CRH-41 (100 pM-1 mM), forskolin (1 nM-1 mM) and an L-Ca2+-channel opener BAY K8644 (100 pM-1 microM) initiated concentration dependent increases in immunoreactive- (ir-) ACTH release from the pituitary cells that were reduced (P < 0.01) by preincubation with dexamethasone (100 nM, 2.5 h). The inhibitory effects of the steroid were reversed by the LC1 antisense ODN (50 nM, P < 0.01), whereas the LC1 sense and scrambled control sequences (50 nM) were both ineffective in this respect (P > 0.05). The results add further support to the view that the acute inhibitory effects of glucocorticoids on the secretion of ACTH by the pituitary gland are dependent on the generation of lipocortin 1. PMID- 9202236 TI - Estrogen regulation of human osteoblastic cell proliferation and differentiation. AB - Estrogen (E2) has been shown to prevent bone loss among postmenopausal women. The molecular mechanism(s) by which this is accomplished is not clear. The discovery of E2 receptor (ER) in osteoblasts and osteoclasts has implicated these cells as direct targets for E2. Previous studies on the effects of E2 on osteoblastic cells in vitro or in organ culture present conflicting results, possibly due to heterogeneity in cell types, stage of differentiation, ER levels, and/or species differences. The effects of E2 on gene expression during various stages of human osteoblast cell differentiation has not been investigated extensively. In this study we employed a newly developed human fetal osteoblastic cell line (hFOB/ER9) that contains high levels of ER to examine the effects of E2 on osteoblast proliferation and differentiation. The basal levels and E2 effects on the expression of various extracellular matrix proteins were also characterized throughout different stages of differentiation. These stages include a proliferative/relatively undifferentiated stage (day 6), a matrix maturation stage (days 10-14), and a mineralization/calcified nodule stage (day 18). During the stage of rapid cell proliferation, E2 treatment of hFOB/ER9 cells resulted in a dose-dependent decrease in [3H]thymidine incorporation to a maximum of 72% compared to the vehicle control value. Treatment of hFOB/ER9 cells with 10(-9) M E2 for 48 h resulted in an increase in alkaline phosphatase (AP) activity throughout cell differentiation. The magnitude of AP induction varied from approximately 200-500%. In contrast, E2 decreased osteocalcin protein levels to a minimum of 54% compared to the vehicle control value. The steady state messenger RNA levels for AP increased and osteocalcin decreased after E2 treatment, similar to the responses observed at the protein level. At all stages, there was little or no effect of E2 on type I collagen protein levels or osteonectin steady state messenger RNA levels. The E2 responses on hFOB/ER9 cell matrix protein expression and cell proliferation were mediated through the ER, as cultures cotreated with a 100-fold molar excess of a type II anti-E2 (ICI 182,780) abrogated these effects. These results support the hypothesis that E2 does have an effect on osteoblastic differentiation by decreasing hFOB/ER9 cell proliferation and differentially regulating extracellular matrix expression. PMID- 9202237 TI - Inhibin interferes with activin signaling at the level of the activin receptor complex in Chinese hamster ovary cells. AB - To gain more insight in the mechanism of action of inhibin, we studied the effect of inhibin on activin signaling in Chinese hamster ovary cells. Inhibin specifically counteracted activin-induced expression of a plasminogen activator inhibitor 1 promoter element (3TP) and of the junB gene, but was ineffective when the responses were induced by transforming growth factor-beta. This indicates that inhibin acts only on the activin-specific part of these signaling cascades. Using a constitutively active activin type IB receptor we determined whether inhibin acted at the level of the activin-receptor complex or downstream of it. The mutant activin receptor stimulated the expression of the 3TP promoter in the absence of activin. This stimulation was insensitive to inhibin, indicating that inhibin acts exclusively at or upstream of this activin type I receptor. In addition, competition studies using labeled activin showed that inhibin displaced activin from the activin type II receptors, especially from the activin type IIB receptor, but not from the type I receptors. In conclusion, these data show that in Chinese hamster ovary cells inhibin acts directly at the activin receptor complex, most likely through displacement of activin from the activin type II receptor. PMID- 9202238 TI - Liver-specific expression of human insulin-like growth factor binding protein-1 in transgenic mice: repercussions on reproduction, ante- and perinatal mortality and postnatal growth. AB - Study of the in vivo functions of the insulin-like growth factor binding proteins (IGFBPs) is complicated by their variety (six molecular species) and the differences in their expression related to tissue of origin and stage of development. To investigate the physiological role of IGFBP-1 in the bloodstream, we induced hepatic overexpression of IGFBP-1 in transgenic mice, placing human IGFBP-1 (hIGFBP-1) cDNA under the control of the alpha1-antitrypsin promoter so as to obtain liver-specific expression. Five transgenic founder mice were raised, only two of which (lines 124 and 149) produced transgenic offspring. Northern blotting revealed transgene expression exclusively in the liver during fetal life and unchanged through to adulthood, whereas expression of the endogenous gene was undetectable beyond 10-15 days postnatally. hIGFBP-1 was detected by western immunoblotting in the plasma of transgenic mice and IRMAs yielded mean concentrations of 2.41 +/- 0.33 ng/ml and 13.69 +/- 1.42 ng/ml in homozygous animals of lines 124 and 149, respectively. In the latter, IGFBP-1 levels were distinctly higher than in heterozygotes (2.99 +/- 0.39 ng/ml), P < 0.0001. These levels remained stable in each given animal and did not change with age. Plasma concentrations of IGF-I measured in line 149 exhibited the well-known profile of an increase from birth up to puberty. Values for heterozygotes were similar to those for wild-type mice, with adult levels (544 +/- 98 ng/ml) slightly below those of controls (630 +/- 56 ng/ml), P < 0.05. In homozygotes they were distinctly lower, with adult levels of 370 +/- 75 ng/ml, P = 0.001. In heterozygous and homozygous adults, there was a negative correlation between IGF I and IGFBP-1 concentrations (r = 0.8, P < 0.0001), suggesting a link between transgene expression and IGF-I levels. Study of body weight gain in line 149 revealed growth retardation within the first weeks after birth, which was marked in homozygous males and females (P < 0.001) but also present in heterozygous males (P = 0.002), indicating some relationship with transgene expression. In addition, body weight in adult mice was negatively correlated to plasma concentrations of IGFBP-1 (r = 0.7, P < 0.0001). Reproductive function also appeared to be severely affected, especially in homozygous females: mating that failed to result in pregnancy in half of the homozygous females crossed with nontransgenic males, suggestive of impaired fertilization or implantation; interrupted or prolonged pregnancies with fetal and neonatal death. Litter size was reduced in transgenic females (by about half in homozygotes) and in nontransgenic females mated with homozygous males, resulting from pre- or neonatal mortality. Moreover, deaths occurred within the first 5 days of life, with an incidence of approximately 50% in the litters of homozygous females, 12 18% among heterozygotes mated with nontransgenic or heterozygous males, respectively, and 30% among those mated with homozygous males. These results, suggesting that fetal transgene expression largely accounted for ante- and perinatal mortality, were confirmed by the predominance of homozygotes among those that could be analyzed genetically. Similarly impaired reproductive function was seen in line 124, but to a lesser degree. Although the mechanisms responsible for these disorders remain to be determined, our results indicate that permanent and uncontrolled hepatic expression of IGFBP-1, even at low levels, affects fertility in females and both ante- and postnatal development. PMID- 9202239 TI - Oxoreductase and dehydrogenase activities of the human and rat 11beta hydroxysteroid dehydrogenase type 2 enzyme. AB - The 11beta-hydroxysteroid dehydrogenase type 2 enzyme (11betaHSD2) metabolizes glucocorticoids into their inactive 11-keto metabolites. Although the type 1 enzyme (11betaHSD1) displays both oxidative and reductive activity, to date 11betaHSD2 has been shown to have dehydrogenase activity only. In this study we compared both dehydrogenase and reductase characteristics of the cloned rat 11betaHSD1 and rat and human 11betaHSD2 for three different 11-hydroxysteroid substrates, cortisol (F), corticosterone (B), and dexamethasone (Dex), and the corresponding 11-keto metabolites, cortisone (E), 11-dehydrocorticosterone (A), and 11-dehydrodexamethasone (DH-Dex), respectively. In cell homogenates expressing either the rat or the human 11betaHSD2, the relative potency for the dehydrogenase reaction was B > F > Dex. Although there was no reduction of A or E, DH-Dex was readily converted to Dex with an equilibrium far on the side of the 11-hydroxy metabolite. DH-Dex reduction in homogenates was inhibited by both glycyrrhetinic acid and carbenoxolone, with a 50% inhibition at 80 and 100 nM, respectively. In intact cells transfected with rat 11betaHSD1, the equilibrium was on the reductase side for all substrates. Dehydrogenation of B or F was more potent with rat 11betaHSD2 than with rat 11betaHSD1. There was no detectable 11betaHSD1 oxidation of Dex. These data indicate that both the cloned human and rat 11betaHSD2 reduce DH-Dex and do this more readily than they oxidize Dex. Thus, 11betaHSD2 seems also to be a bidirectional enzyme, although no reduction of the physiological compounds A and E was observed. PMID- 9202240 TI - Anabolic effects of 1,25-dihydroxyvitamin D3 on osteoblasts are enhanced by vascular endothelial growth factor produced by osteoblasts and by growth factors produced by endothelial cells. AB - Human osteoblast-like cells (HOB) produce vascular endothelial growth factor (VEGF), the steady state level of which is stimulated by 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3]. As osteoblasts and endothelial cells are proximally located in skeletal tissue, we investigated the anabolic effects of 1,25-(OH)2D3 and VEGF on HOB cocultured with endothelial cells. When HOB with high alkaline phosphatase (Al-P) activity and human umbilical vein endothelial cells (HUVEC) with little activity were cultured together, Al-P activity increased, accompanied by an increase in cell number. When HOB and HUVEC were cultured separately, 1,25 (OH)2D3 did not directly stimulate [3H]thymidine incorporation into HUVEC, but stimulated it in the presence of HOB. VEGF did not directly stimulate the Al-P activity of HOB but stimulated it in the presence of HUVEC. The conditioned medium of HOB stimulated the proliferation of HUVEC, and this was partially blocked by anti-VEGF antibody. Conversely, the conditioned medium of HUVEC increased Al-P activity and [3H]thymidine incorporation into HOB, and this was partially blocked by antiinsulin-like growth factor I antibody and BQ-123, a specific antagonist of the endothelin-1 (ET-1) receptor. 1,25-(OH)2D3 stimulated the release of VEGF and ET-1 from HOB and HUVEC, respectively. Furthermore, the 1,25-(OH)2D3-induced release of VEGF was enhanced in HOB cocultured with HUVEC. A quantitative reverse transcription-PCR study revealed that genes for VEGF receptors (Flt-1 and KDR) were expressed in HUVEC, but not in HOB, and that 1,25 (OH)2D3 increased the levels of expression of VEGF receptor genes in endothelial cells only when cocultured with HOB. In summary, we demonstrated that 1,25 (OH)2D3 exerts an anabolic effect on osteoblasts by enhancing their production of VEGF, which stimulates its receptors on endothelial cells, followed by increased production of osteotropic growth factors, such as insulin-like growth factor I and ET-1. These in vitro findings suggest that the VEGF/VEGF receptor system may be involved in both bone formation and bone remodeling in vivo. PMID- 9202241 TI - Selection of the dominant follicle in cattle occurs in the absence of differences in the expression of messenger ribonucleic acid for gonadotropin receptors. AB - Mechanisms that allow selection of a dominant ovarian follicle from a cohort of growing follicles are unknown. Large healthy, estrogen-active follicles contain more LH receptors than atretic estrogen-inactive follicles, and levels of messenger RNA (mRNA) for LH receptor increase in the granulosa cells of dominant follicles as growth progresses. The aim of the present study was to test the hypothesis that changes in the temporal pattern of expression of mRNA for LH and FSH receptors are associated with selection of dominant follicles in cattle. Based on size, the dominant and two largest subordinate follicles were collected from the ovaries of heifers on days 2 (n = 3) or 3 (n = 3) of a follicular wave. On day 2, the dominant follicle was 1 mm larger than the largest subordinate follicle, but by day 3 of the wave the dominant follicle was 2-4 mm larger than the largest subordinate. Follicular fluid concentrations of estradiol and estradiol secretion in vitro by pieces of follicle wall (granulosa and theca cells) were greatest by the dominant compared with the subordinate follicles (P < 0.05). These data indicate that selection of a dominant follicle had occurred by the second day of the follicular wave. By in situ hybridization, mRNAs for LH and FSH receptors, P450 aromatase and P450 17alpha-hydroxylase (17alpha-OH) were localized in frozen sections from each follicle. The expression of mRNA for LH receptor in granulosa cells was always at or near background and was not different between days or follicle types (P = 0.63). In contrast, the expression of mRNA for LH receptor in theca cells of the same sections was readily detectable; there was no difference between follicle types on the second day of the follicular wave, but by the third day expression in the subordinate follicles had decreased (P < 0.05). The expression of mRNA for FSH receptor was highest in granulosa cells of dominant follicles collected on day 3 of the follicular wave (P < 0.05) and was not different between dominant and subordinate follicles on day 2 of the wave (P > 0.05). The expression of mRNA for aromatase in granulosa cells was similar (P > 0.05) between the dominant follicles on days 2 and 3 and the largest subordinate follicle on day 2 of the follicular wave and was much lower in the remaining follicles (P < 0.01). On day 2 of the wave, the expression of mRNA for 17alpha-OH was not different between the dominant and subordinate follicles, but by day 3 the dominant follicles had more mRNA for 17alpha-OH than the subordinate follicles (P < 0.05). These data show that the dominant follicle had been selected by the second day of the follicular wave (based on diameter and estradiol secretion) and that selection occurred in the absence of detectable levels of mRNA for LH receptor in the granulosa cells or differences between dominant and subordinate follicles in mRNA for LH receptor in theca cells or FSH receptor in granulosa cells. However, the divergent pattern of growth between dominant and subordinate follicles (after follicle selection) was associated with higher levels of mRNA for gonadotropin receptors and steroidogenic enzymes in dominant compared with subordinate follicles. Therefore, selection of the dominant follicle in cattle does not appear to involve the regulation of expression of mRNA for gonadotropin receptors, although such regulation may be important at other stages of differentiation of the dominant follicle. PMID- 9202242 TI - Insulin-like growth factor binding protein-5 binds to plasminogen activator inhibitor-I. AB - Insulin-like growth factor binding protein-5 (IGFBP-5) has been shown to bind to the extracellular matrix (ECM) of both fibroblasts and smooth muscle cells. The ECM-IGFBP-5 interaction is mediated in part by binding to heparan sulfate containing proteoglycans. Because proteoglycans may not be the only components of ECM that bind to IGFBP-5, we have determined its ability to bind to other ECM proteins. When a partially purified mixture of the proteins that were present in fibroblast conditioned medium was purified by IGFBP-5 affinity chromatography, a 55-kDa protein was eluted. Amino acid sequencing of the amino terminal 28 amino acids showed that it was human plasminogen activator inhibitor-1 (PAI-1). To determine if this interaction was specific, purified human PAI-1 was incubated with IGFBP-5 and the IGFBP-5/PAI-1 complex immunoprecipitated with anti-PAI-1 antiserum. When the precipitate was analyzed by immunoblotting using anti-IGFBP-5 antiserum, the intensity of the IGFBP-5 band was substantially increased compared with controls that did not contain human PAI-1. A synthetic IGFBP-5 peptide that contained the amino acid sequence between positions 201 and 218 inhibited IGFBP 5/PAI-1 interaction. Coincubation of IGFBP-5 mutants that contained substitutions for specific basic residues located between positions 201 and 218 with PAI-1 indicated that some of these amino acids were important for binding. Two mutants that contained neutral substitutions for specific basic amino acids within the glycosaminoglycan binding domain had reduced binding to PAI-1. In contrast, three other mutants that also had substitutions for charged residues in the same region had no reduction in binding. Heparin and heparan sulfate inhibited the IGFBP 5/PAI-1 interaction; however, several other glycosaminoglycans had no effect. PAI 1 was determined to be an important ECM component for binding because approximately 27% of total ECM binding could be inhibited with anti-PAI-1 antiserum. Competitive binding studies with unlabeled IGFBP-5 showed that the dissociation constant of PAI-1 for IGFBP-5 was 9.1 x 10(-8) M. In summary, IGFBP 5 binds specifically to plasminogen activator inhibitor-1. Because this is present in the extracellular matrix of several cell types, it may be one of the important binding components of ECM. PAI-1 binding partially protects IGFBP-5 from proteolysis, suggesting that it is one of the ECM components that is involved in mediating this effect. PMID- 9202244 TI - The type 2 and type 3 iodothyronine deiodinases play important roles in coordinating development in Rana catesbeiana tadpoles. AB - In developing Rana catesbeiana tadpoles, the timing of the thyroid hormone (TH) dependent metamorphic responses varies markedly among tissues. Yet at any one time these tissues are exposed to the same plasma concentration of TH, suggesting that TH action is regulated in part at the level of the peripheral tissues. A major factor in TH action is the intracellular level of the active TH, T3. This level is dependent not only on the plasma concentration of TH (mostly T4) but also on the intracellular activities of the type 2 5'-deiodinase (D2) and the type 3 5-deiodinase (D3), which are responsible, respectively, for generating and degrading T3. (D1 is not present in this species.) To determine whether differential expression of D2 and D3 among tissues could be a significant factor in the coordination of metamorphic events, the ontogenic profiles of the two enzyme activities and corresponding messenger RNA levels in most tissues of R. catesbeiana tadpoles have been documented. The profiles of D2 expression in tail, hindlimb, forelimb, intestine, skin, and eye differed markedly at both activity and messenger RNA levels, but it was notable that expression was invariably highest in a given tissue at the time of its major metamorphic change. D2 expression was very low in brain and heart and did not vary during development. D2 was not expressed in liver, kidney, or red blood cells. With the exception of red blood cells, D3 expression was detected in all tissues studied. Furthermore, it was evident that in tissues that expressed both deiodinase genes, the two expression profiles were comparable, indicating a potential for tight control of intracellular T3 levels. Direct evidence of the importance of the intracellular conversion of T4 to T3 for TH-dependent metamorphic events was obtained in tadpoles in which endogenous TH synthesis was blocked with methimazole, and the activities of D2 and D3 were inhibited by iopanoic acid. This treatment inhibited metamorphosis. The inhibition could be overcome by the concomitant administration of replacement levels of T3, but not T4. These results strongly support the view that coordinated development in amphibia depends in part on the tissue-specific expression patterns of the D2 and D3 genes, which ensure that the requisite level of intracellular T3 is attained in a given tissue, regardless of the current level of circulating TH, at the appropriate stage of metamorphosis. PMID- 9202243 TI - Tyrosine residues in the C-terminal domain of the insulin-like growth factor-I receptor mediate mitogenic and tumorigenic signals. AB - We investigated cellular proliferation, the transforming activity, and activation of known signal transduction pathways in NIH-3T3 cells stably expressing insulin like growth factor-I receptors (IGF-IRs) with amino acid substitutions in the carboxy(C)-terminal domain. The mutant receptors contained substitutions of both tyrosines 1250 and 1251 with phenylalanine and histidine (amino acids present in the analogous positions in the insulin receptor), as well as phenylalanine 1310 replaced by tyrosine (IsY clones) to resemble the placement of tyrosine residues in the C-terminal domain of the insulin receptor. As a control for the IsY clones, a second mutant receptor was expressed with a substitution of phenylalanine 1310 with tyrosine only (DBY clones). Clones expressing IGF-IRs with the IsY substitutions had a significantly slower rate of growth compared with cells expressing an equivalent number of wild-type IGF-IRs (NWT). In contrast, the DBY clones showed relatively normal growth rates. Cells with wild type IGF-IR demonstrated a transformed phenotype in soft agar assays. The IsY clones lost the transforming ability of the wild type IGF-IR, whereas DBY clones formed colonies. IGF-I-stimulated autophosphorylation of the IGF-IR and tyrosine phosphorylation of IRS-1 and SHC, known substrates in the IGF-IR signal transduction pathway, were studied. Mutated IGF-IRs (IsY and DBY) did not alter the IGF-I-induced tyrosine phosphorylation of these proteins. Furthermore, the mutated IGF-IRs did not alter Grb2 association with phosphorylated IRS-1 and SHC. IGF-I stimulation of Crk-II phosphorylation, a novel substrate of the IGF-IR, was similar in cells expressing mutated and wild-type IGF-IRs. IGF-I-induced activation of phosphatidylinositol (PI) 3'-kinase was equivalent in cells expressing either mutant or wild-type IGF-IRs. These data suggest that the IGF-IR mediates, at least in part, cellular proliferation and increased transforming ability through its C-terminal domain. The exact postreceptor signaling pathway(s) involved have yet to be fully elucidated. PMID- 9202245 TI - Differential regulation of two uridine diphospho-glucuronosyltransferases, UGT2B15 and UGT2B17, in human prostate LNCaP cells. AB - Although androgens are important regulators in the prostate, other effectors such as growth factors may also act to maintain normal function of the gland. Human prostate and human prostate cancer LNCaP cells express steroid conjugating uridine diphospho-glucuronosyltransferase (UGT) enzymes, and it was shown that the level of UGT activities and transcripts is down-regulated by androgens, especially dihydrotestosterone (DHT). In the present study, we examined the interaction between androgen, epidermal growth factor (EGF), and steroid UGT enzymes. The formation of DHT glucuronide (DHT-G) was inhibited by 47% when LNCaP cells were treated for 6 days with 10 ng/ml of EGF. Northern blot analysis also demonstrated a decrease in the steady-state level of UGT2B transcripts. Treatment with both DHT (0.5 nM) and EGF (10 ng/ml) caused a greater decrease of DHT glucuronidation and UGT2B messenger RNA levels than when the cells were treated with either compound alone. RNase protection assays showed that treatment with DHT and EGF caused a specific decrease of UGT2B17 transcript in LNCaP cells treated; however, the level of UGT2B15 messenger RNA was not affected. As well, Western blot analysis demonstrated a diminution of UGT2B17 protein level in response to DHT and EGF. These results demonstrate a differential regulation of different isoforms of steroid conjugating UGTs present in human prostate LNCaP cells. UGT2B17 was shown to be more labile than UGT2B15, indicating that regulation of UGT2B17 expression would lead to a more rapid change in the level of glucuronidated steroids. PMID- 9202246 TI - Characterization of two distinct intracellular GLUT4 membrane populations in muscle fiber. Differential protein composition and sensitivity to insulin. AB - A major objective for the understanding of muscle glucose disposal is the elucidation of the intracellular trafficking pathway of GLUT4 glucose carriers in the muscle fiber. In this report, we provide functional and biochemical characterization of two distinct intracellular GLUT4 vesicle pools obtained from rat skeletal muscle. The two pools showed a differential response to insulin; thus, one showed a marked decrease in GLUT4 levels but the other did not. They also showed a markedly different protein composition as detected by quantitative vesicle immunoisolation analysis. The GLUT4 pool showing no response to insulin contained SCAMP proteins and the vSNARE proteins VAMP2 and cellubrevin, whereas only VAMP2 was found in the insulin-recruitable GLUT4 pool. SDS-PAGE and further silver staining of the immunoprecipitates revealed discrete polypeptide bands associated to the insulin-sensitive pool, and all these polypeptide bands were found in the insulin-insensitive population. Furthermore, some polypeptide bands were exclusive to the insulin-insensitive population. The presence of cellubrevin and SCAMP proteins, endosomal markers, suggest that the insulin-insensitive GLUT4 membrane population belongs to an endosomal compartment. In addition, we favor the view that the insulin-sensitive GLUT4 membrane pool is segregated from the endosomal GLUT4 population and is undergoes exocytosis to the cell surface in response to insulin. Intracellular GLUT4 membranes obtained from skeletal muscle contain cellubrevin, and VAMP2 and GLUT4-vesicles from cardiomyocytes also contain cellubrevin. This suggests that vSNARE proteins are key constituents of GLUT4 vesicles. The presence of the tSNARE protein SNAP25 in skeletal muscle membranes and SNAP25 and syntaxin 1A and syntaxin 1B in cardiomyocyte plasma membranes further suggest a role of the SNAREs in GLUT4 trafficking in muscle. PMID- 9202248 TI - Spatial and temporal changes in the insulin-like growth factor (IGF) axis indicate autocrine/paracrine actions of IGF-I within wounds of the rat brain. AB - A precise role for insulin-like growth factors (IGFs), IGF-binding proteins (IGFBPs), and IGF-receptors (IGF-Rs) in damaged central nervous system (CNS) tissue has not been elucidated, although their expression in the ischemic brain has been demonstrated. However, little is known of IGF responses after CNS trauma. In this study, we have used ribonuclease protection assay, in situ hybridization, and immunohistochemistry to demonstrate that IGF-I, IGFBPs, and IGF-1R expression alters in response to a penetrating CNS injury. Within penetrant cerebral wounds in the acute phase of the response (1-7 days post lesion; dpl), increased levels of IGF-I, IGFBP-1, -2, -3, -6, and IGF-1R protein were localized to injury responsive astrocytes, neurons and cells of the monocyte lineage. IGF-I, IGFBP-2, and 3 showed a congruency in sites of messenger RNA (mRNA) and peptide expression, with IGF-I and IGFBP-2 mRNA expression predominating. IGF-I, IGFBP-1, and IGFBP-3 protein were also associated with the microvascular endothelium, which was accompanied by increased levels of IGFBP-3 mRNA. These early changes in IGFBP expression probably facilitate IGF-I action. Later in the wounding response (7-14 dpl), the expression of IGFBP-4 and IGFBP-5 peaked within astrocytes and neurons, with IGFBP-5 mRNA being specifically localized to the glia limitans within the wound, suggesting an inhibitory role for these proteins, down-regulating the effects of IGF-I chronically. Our evidence suggests that within penetrating CNS wounds, IGF-I acts in an autocrine/paracrine manner to regulate cellular responses, with its spatial and temporal availability being modulated by the differential presence of stimulatory vs. inhibitory IGFBPs. PMID- 9202247 TI - Prolactin (PRL) receptors are colocalized in dopaminergic neurons in fetal hypothalamic cell cultures: effect of PRL on tyrosine hydroxylase activity. AB - This study examined the responsiveness of dopaminergic neurons to PRL and the expression of PRL receptors in fetal hypothalamic cells. Hypothalamic cells were cultured in medium containing 5 or 25 mM potassium (K+) with or without 5% FBS. Rat PRL (rPRL) treatment (10-1000 ng/ml) for 10 days increased tyrosine hydroxylase (TH) activity 1.6- to 1.8-fold in dopaminergic neurons cultured in serum-containing medium with 25 mM K+, but not in defined medium or any medium with 5 mM K+. The rPRL-induced increase in TH activity was observed at 10-1000 ng/ml after both 1 and 10 days of rPRL treatment, whereas 1 ng/ml was not effective. TH activity was not altered after 1-12 h of rPRL treatment (100 ng/ml), but was increased 1.4-fold after 1-3 days and 1.8-fold after 5-10 days. The colocalization of PRL receptors and TH was evaluated by double labeled immunocytochemistry. PRL receptor immunostaining was observed in most TH immunoreactive cells cultured in either defined or serum-containing medium with or without 10 days of rPRL treatment (100 ng/ml). As assessed by reverse transcriptase-PCR, the long form, but not the short form, of the PRL receptor was expressed in the hypothalamic cells regardless of medium composition, similar to the expression pattern in adult mediobasal hypothalamus from ovariectomized rats. These data indicate that a factor present in FBS imparts PRL responsiveness to hypothalamic dopaminergic neurons in vitro. The effective PRL concentrations and the time course for PRL's action in vitro are within the physiological range in vivo. The colocalization of PRL receptor in dopaminergic neurons provides anatomical evidence for a direct effect of PRL, with the long form of the PRL receptor being the predominant form in the hypothalamic cells. PMID- 9202250 TI - The uterine myometrium is a target for increased levels of activin A during pregnancy. AB - Activin A is a dimeric protein hormone that regulates numerous cellular functions. A clear physiological role for this molecule in pregnancy is suggested by previous studies in the human, wherein activin A rises dramatically as women approach parturition. To determine whether the rodent is a suitable animal model for further studies of activin action during pregnancy, the serum concentration of activin A was measured in pregnant rats. Activin A was detected in the serum of pregnant rats, beginning on day 12, and the serum concentration rose progressively through gestation (22-fold) and dramatically (140-fold) in labor. The potential target tissues for circulating activin were then identified in two ways. First, iodinated activin was injected into pregnant rats, and the tissues targeted by labeled ligand were identified in vivo. A tissue targeted by activin A in the pregnant rat was the uterine myometrium. To determine the ligand specificity of the uterine myometrial cells, the uteri of pregnant rats were collected and analyzed by in situ ligand binding. 125I-activin A binding was specific for the uterus myometrium, and the ligand binding was competed by unlabeled activin A but not by inhibin A. This result suggests that the receptor in this tissue compartment is an activin-specific receptor. The production of abundant activin A and the ability of exogenous ligand to target the myometrium of the uterus provides a pathway by which activin could regulate uterine function during pregnancy. PMID- 9202249 TI - Retinoic acid and alcohol/retinol dehydrogenase in the mouse adrenal gland: a potential endocrine source of retinoic acid during development. AB - Retinoid signaling requires the conversion of retinol to retinoic acid by a two step process, the first of which can be catalyzed in vitro by class I and class IV alcohol dehydrogenases (ADH). These enzymes may participate in local retinoic acid synthesis in some target tissues, although other studies suggest retinoic acid may also be supplied to tissues via the bloodstream, much like an endocrine hormone. Here we have analyzed the expression of these two ADHs as well as retinoic acid production in the adrenal gland, an organ known to be an endocrine source of other hormones. In situ hybridization revealed high levels of both class I and class IV ADH messenger RNAs in adrenal glands of 16.5-day mouse embryos and adults. Class I ADH protein was immunohistochemically detected in embryonic and adult adrenal glands, the latter primarily in the zona fasiculata of the cortex. Abundant class IV ADH protein was detected in the embryonic adrenal as well as in the zona glomerulosa and zona fasiculata of the adult adrenal cortex. Interestingly, class IV ADH protein was found in only a subset of adult cortical cells arranged in radial columns, thus providing further evidence for centripetal cell migration during adrenocortical differentiation. Using a retinoic acid bioassay, adrenal glands from 16.5 day embryos were found to have significantly higher levels of retinoic acid than embryonic liver. The adult adrenal was found to have approximately 15.5 pmol/g of retinoic acid, whereas the adult liver had 24.8 pmol/g, and brain, heart, and spleen each had less than 1.0 pmol/g. Because previous findings indicate that the adrenal gland is not a retinoid target tissue, our detection of both alcohol/retinol dehydrogenases and significant amounts of retinoic acid in this organ suggests that it functions as a potential endocrine source of this hormone during mouse development. PMID- 9202251 TI - Osteopontin expression by osteoclast and osteoblast progenitors in the murine bone marrow: demonstration of its requirement for osteoclastogenesis and its increase after ovariectomy. AB - Osteoclast development requires cell-to-cell contact between hematopoietic osteoclast progenitors and bone marrow stromal/osteoblastic support cells. Based on this, we hypothesized that osteopontin, an adhesion protein produced by osteoclasts and osteoblasts, plays a role in osteoclastogenesis. Using in situ hybridization, we demonstrate that cells expressing the osteopontin messenger RNA (mRNA) appear after 3 days of culturing murine bone marrow cells. The number of these cells increases thereafter, reaching a peak on day 5. In the same cultures, cells expressing alkaline phosphatase (AP) or tartrate resistant acid phosphatase (TRAP), phenotypic markers for osteoblastic and osteoclast-like cells, respectively, appeared subsequent to the appearance of the osteopontin-positive cells. By means of a combination of in situ hybridization and histostaining, it was shown that the osteopontin mRNA was localized in 30-50% of the AP-positive or the TRAP-positive, as well as in nonspecific esterase (NSE)-positive, cells. The number of cells expressing both the osteopontin mRNA and either one of the three phenotypic markers was significantly increased in bone marrow cultures from estrogen-deficient mice, as compared with controls. Conversely, the number of all three populations of double positive cells was decreased in cultures treated with a specific antimouse rabbit osteopontin antibody or an RGD peptide. These findings indicate that osteopontin is expressed during the early stages of the differentiation of osteoclast and osteoblast progenitors in the bone marrow and that its cell adhesion properties are required for osteoclastogenesis. PMID- 9202252 TI - Regulation of apoptosis in uterine leiomyomata. AB - Tumors developing from hormone-dependent tissues, such as the breast and prostate, have been successfully treated in the clinic by methods of hormone ablation, and the resulting tumor regression has been shown to occur at least in part by the process of apoptosis. The growth of leiomyomas arising from uterine smooth muscle cells is similarly modulated by circulating steroid hormones and has been associated with periods of increased estrogen secretion. The inhibition of ovarian hormone production by endocrine therapy often results in the regression of these tumors, but the role of apoptosis in this process has not been elucidated. Using cell lines derived from the Eker rat model of uterine leiomyoma, we have investigated the mechanism of growth inhibition by estrogen deprivation. Estrogen-depleted medium and the antiestrogen tamoxifen significantly reduced cell numbers in culture and arrested cell proliferation, but did not induce apoptosis. However, the presence of an intact apoptotic pathway was demonstrated in these cells by serum starvation. In vivo data were in agreement with in vitro results, which showed that tamoxifen treatment does not change the apoptotic rate of leiomyoma tissues. Therefore, growth modulation of leiomyomas by hormone deprivation occurs via mechanisms independent of apoptosis, indicating a fundamental difference in the response of leiomyomas to hormone deprivation from that of tumors of the breast and prostate. These data suggest that creation of a hypoestrogenic milieu within leiomyomas reduces tumor volume without inducing a concomitant increase in the rate of apoptosis, which may be responsible for the limited effectiveness of currently available medicinal therapies. PMID- 9202253 TI - Immunohistochemical localization of somatostatin receptor SST2A in the rat pancreas. AB - Somatostatin (SRIF) acts on specific membrane receptors to inhibit exocrine and endocrine pancreatic functions. Five SRIF receptor genes have been cloned, producing six receptor proteins (sst-s). We used a recently developed antibody to localize the sst2A splice variant in the rat pancreas. Western blots identified the sst2A receptor as an 90 kDa glycosylated protein in pancreatic tissue. In tyramide-amplified immunostainings all acinar cells, and the glucagon and pancreatic polypeptide immunoreactive cells (A and PP, respectively) were intensely labeled for sst2A, while no signal was detected in SRIF producing (D) cells. A very few insulin immunoreactive (B) cells were also labeled for sst2A, but the signal in these cells was lower than in exocrine, A or PP cells. Absorption of the sst2A antibody with the receptor peptide abolished specific staining in both immunoblots and tissue sections (negative control). These studies are the first to localize any SRIF receptor subtype in the rat pancreas. The specific localization of sst2A receptor in acinar, A and PP cells if confirmed in humans, would suggest that subtype specific analogs will be useful for the therapeutic regulation of exocrine and/or endocrine pancreatic secretion. PMID- 9202254 TI - Immunization of male bonnet monkeys (M. radiata) with a recombinant FSH receptor preparation affects testicular function and fertility. AB - Immunization of proven fertile adult male monkeys (n=3) with a recombinant FSH receptor protein preparation (oFSHR-P) (representing amino acids 1-134 of the extracellular domain of the receptor Mr approximately 15KDa) resulted in production of receptor blocking antibodies. The ability of the antibody to bind a particulate FSH receptor preparation and receptors in intact granulosa cells was markedly (by 30-80%) inhibited by FSH. Serum T levels and LH receptor function following immunization remained unchanged. The immunized monkeys showed a 50% reduction (p<0.001) in transformation of spermatogonia(2C) to primary spermatocytes (4C) as determined by flow cytometry and the 4C:2C ratio showed a correlative change (R 0.81, p<0.0007) with reduction in fertility index (sperm counts X motility score). Breeding studies indicated that monkeys became infertile between 242-368 days of immunization when the fertility index was in the range of 123+/-76 to 354+/-42 (compared to a value of 1602+/-384 on day 0). As the effects observed are near identical to that seen following immunization with FSH it is suggestive that oFSHR-P can substitute for FSH in the development of a contraceptive vaccine. PMID- 9202255 TI - The 16-kDa proteolytic fragment of insulin-like growth factor (IGF) binding protein-3 inhibits the mitogenic action of fibroblast growth factor on mouse fibroblasts with a targeted disruption of the type 1 IGF receptor gene. AB - We previously reported that a 16-kDa proteolytic fragment of IGF Binding Protein 3 (IGFBP-3), which is devoid of affinity for IGFs, inhibits the mitogenic effects of IGF-I on chick embryo fibroblasts. Here, we set out to determine if the fragment had biological effects on fibroblasts from mouse embryos homozygous for a targeted disruption of the Type 1 IGF receptor gene. In the cell clone used, bFGF (but not IGF, EGF or PDGF) was mitogenic in serum-free medium, increasing 14C-thymidine uptake by a factor of 10-15 within 24 hours and doubling cell proliferation. The 16-kDa fragment, isolated by HPLC following limited proteolysis of recombinant human (rh) IGFBP-3 by plasmin, in both assays dose dependently (20 to 100 ng/ml) inhibited (up to 100%) maximal stimulation induced by 25 ng/ml bFGF, whereas intact IGFBP-3 had virtually no effect. Similar results were obtained with control wild-type cells. In the latter, the mitogenic activity of 1% fetal calf serum (equal to that of 25 ng/ml bFGF) was inhibited by only 25 30% by 100 ng/ml 16-kDa fragment or 200 ng/ml rhIGFBP-3. This agrees with an antagonistic action, affecting the mitogenic activity of serum that is attributable to IGFs. The 16-kDa IGFBP-3 fragment therefore appears to be a potent inhibitor of mitogenic signals resulting from activation of both the type 1 IGF and FGF receptors. PMID- 9202256 TI - Decreased expression of murine PPARgamma in adipose tissue during endotoxemia. AB - Infection-induced hyperlipidemia develops due to a combination of factors, one of which is decreased clearance of lipids from the bloodstream due to depressed synthesis of lipoprotein lipase (LPL). Recently, the peroxisome proliferator activated receptors (PPARs) have been shown to be important in the regulation of LPL, particularly PPARgamma. PPARgamma and its heterodimerization partner, RXR alpha have been shown to be transcriptional activators of LPL in co-transfection analysis. Therefore, we hypothesized that the decrease in LPL expression during endotoxemia may be a result of depressed PPARgamma expression. In these studies, we examined the effect of endotoxin or its proximal mediator, tumor necrosis factor (TNF), on the expression of PPARgamma in white (WAT) and brown adipose tissue (BAT) in CD-1 mice. We report that treatment with endotoxin, but not TNF, transiently decreased PPARgamma mRNA levels 4 hr after treatment. However, endotoxin or TNF treatment decreased PPARgamma protein levels after 18 hr, which was at a time when LPL mRNA levels were also depressed. These data suggest that decreased PPARgamma expression following endotoxin or TNF treatment may contribute to the hyperlipidemia due to decreased expression of LPL, which would impair triglyceride clearance. PMID- 9202257 TI - Differential expression of nuclear 11beta-hydroxysteroid dehydrogenase type 2 in mineralocorticoid receptor positive and negative tissues. AB - Corticosteroid hormone action is controlled at a pre-receptor level by the activity of two isoforms of 11beta-hydroxysteroid dehydrogenase (11beta-HSD), catalyzing the interconversion of hormonally active cortisol to inactive cortisone. In particular 11beta-HSD2 protects the mineralocorticoid receptor (MR) from glucocorticoid excess, enabling aldosterone to interact with the MR. We have analyzed the subcellular localization of 11beta-HSD2 in relation to the expression of the MR in human colon and placenta. 3H-aldosterone binding studies confirmed expression of the MR in human colon but not term placental trophoblast. Enzyme activity studies and Western blot analyses carried out on subcellular fractions confirmed the presence of 11beta-HSD2 in microsomes. In colon, but not placenta, 11beta-HSD2 was also localized to the microsome-free, nuclear fraction. Protection upon the MR by 11beta-HSD2 in "classical" mineralocorticoid target tissues such as colon can be subserved at both a nuclear and extra-nuclear level. Tissue specific factors are responsible for the subcellular localization of 11beta-HSD2 and we postulate that one such factor may be the MR itself. PMID- 9202258 TI - Cerebral arteriovenous oxygen difference: a predictor of cerebral infarction and outcome in patients with severe head injury. AB - Jugular bulb oxygen monitoring can be used to estimate the adequacy of cerebral blood flow to support cerebral metabolism after severe head injury. In the present study, the authors studied the cerebral arteriovenous oxygen difference (AVDO[2]) before and after treatment in 32 head-injured patients (Glasgow Coma Scale scores < or = 8) to examine the relationships among AVDO and cerebral perfusion pressure (CPP), delayed cerebral infarction, and outcome. Fifteen patients (Group A) underwent craniotomy for hematoma evacuation and 17 (Group B) received mannitol for sustained intracranial hypertension (intracranial pressure > 20 mm Hg, > 10 minutes). Radiographic evidence of delayed cerebral infarction was observed in 14 patients. Overall, 17 patients died or were severely disabled. Cerebral AVDO(2) was elevated before craniotomy or mannitol administration; the mean AVDO(2) for all patients before treatment was 8.6 +/- 1.8 vol%. Following craniotomy or mannitol administration, the AVDO(2) decreased in 27 patients and increased in five patients (mean AVDO(2) 6.2 +/- 2.1 vol% in all patients; 6 +/- 1.9 vol% in Group A; and 6.4 +/- 2.4 vol% in Group B). The mean CPP was 75 +/- 9.8 mm Hg and no relationship with AVDO(2) was demonstrated. Before treatment, the AVDO(2) was not associated with delayed cerebral infarction or outcome. By contrast, a limited improvement in elevated AVDO(2) after craniotomy or mannitol administration was significantly associated with delayed cerebral infarction (Group A: p < 0.001; Group B: p < 0.01). Similarly, a limited improvement in elevated AVDO(2) after treatment was significantly associated with an unfavorable outcome (Group A: p < 0.01; Group B: p < 0.001). In conclusion, these findings strongly indicate that, despite adequate cerebral perfusion, limited improvement in elevated cerebral AVDO(2) after treatment consisting of either craniotomy or mannitol administration may be used to help predict delayed cerebral infarction and poor outcome after traumatic brain injury. PMID- 9202259 TI - Characterization of cerebral hemodynamic phases following severe head trauma: hypoperfusion, hyperemia, and vasospasm. AB - The extent and timing of posttraumatic cerebral hemodynamic disturbances have significant implications for the monitoring and treatment of patients with head injury. This prospective study of cerebral blood flow (CBF) (measured using 133Xe clearance) and transcranial Doppler (TCD) measurements in 125 patients with severe head trauma has defined three distinct hemodynamic phases during the first 2 weeks after injury. The phases are further characterized by measurements of cerebral arteriovenous oxygen difference (AVDO[2]) and cerebral metabolic rate of oxygen (CMRO[2]). Phase I (hypoperfusion phase) occurs on the day of injury (Day 0) and is defined by a low CBF calculated from cerebral clearance curves integrated to 15 minutes (mean CBF 32.3 +/- 2 ml/100 g/minute), normal middle cerebral artery (MCA) velocity (mean V[MCA] 56.7 +/- 2.9 cm/second), normal hemispheric index ([HI], mean HI 1.67 +/- 0.11), and normal AVDO(2) (mean AVDO[2] 5.4 +/- 0.5 vol%). The CMRO, is approximately 50% of normal (mean CMRO(2) 1.77 +/ 0.18 ml/100 g/minute) during this phase and remains depressed during the second and third phases. In Phase II (hyperemia phase, Days 1-3), CBF increases (46.8 +/ 3 ml/100 g/minute), AVDO(2) falls (3.8 +/- 0.1 vol%), V(MCA) rises (86 +/- 3.7 cm/second), and the HI remains less than 3 (2.41 +/- 0.1). In Phase III (vasospasm phase, Days 4-15), there is a fall in CBF (35.7 +/- 3.8 ml/100 g/minute), a further increase in V(MCA) (96.7 +/- 6.3 cm/second), and a pronounced rise in the HI (2.87 +/- 0.22). This is the first study in which CBF, metabolic, and TCD measurements are combined to define the characteristics and time courses of, and to suggest etiological factors for, the distinct cerebral hemodynamic phases that occur after severe craniocerebral trauma. This research is consistent with and builds on the findings of previous investigations and may provide a useful temporal framework for the organization of existing knowledge regarding posttraumatic cerebrovascular and metabolic pathophysiology. PMID- 9202260 TI - A cost-effectiveness analysis of anterior temporal lobectomy for intractable temporal lobe epilepsy. AB - Patients with medically intractable temporal lobe epilepsy are potential candidates for anterior temporal lobectomy (ATL), in which epileptogenic temporal lobe tissue is localized and surgically removed. This surgical approach can eliminate or drastically reduce seizures in the majority of patients. The authors used a decision-analysis model to examine the cost-effectiveness of a surgical evaluation and treatment protocol for medically intractable temporal lobe epilepsy. This model compared a cohort treated with the new protocol with a continuation of their immediate preoperative medical management and projected these differences over the patient's lifetime. The Markov model incorporated postoperative seizure status, patient quality of life, death from surgical and natural causes, discounting, and the direct medical costs associated with outpatient evaluation, hospitalization, surgery, antiepileptic drugs, and lifetime outpatient treatment. The intent-to-treat analysis included patients who underwent evaluation but were not eligible for ATL. Sensitivity analyses were also performed on the variables in the model. Data from the baseline model indicated that evaluation for ATL provided an average of 1.1 additional quality adjusted life years (QALYs) compared with continued medical management, at an additional cost of $29,800. Combining the clinical and economic outcomes yielded a cost-effectiveness ratio of $27,200 per QALY. This value is comparable to other accepted medical or surgical interventions, such as total knee arthroplasty ($16,700/QALY) or coronary artery balloon angioplasty ($40,800/QALY). Sensitivity analyses demonstrate that the results are critically dependent on postoperative seizure status and improvement in quality of life. Although further work is necessary to quantify the improvement in quality of life after epilepsy surgery better, the present data indicate that ATL for treatment of intractable temporal lobe epilepsy is a cost-effective use of medical resources. PMID- 9202261 TI - Albendazole and praziquantel treatment in neurocysticercosis of the fourth ventricle. AB - The purpose of this study was to determine the therapeutic efficacy of albendazole and praziquantel administration in the treatment of neurocysticercosis of the fourth ventricle. The authors report the results obtained in 10 patients with cystic neurocysticercosis of the fourth ventricle who were treated with albendazole at a dosage of 15 mg/kg/day for 2 weeks. Because of the failure of albendazole treatment, two of the patients received an additional course of praziquantel at a dosage of 100 mg/kg/day for 2 weeks. A total of 16 courses of albendazole and two courses of praziquantel were administered to the 10 patients. In eight patients (80%), there was complete disappearance of the cyst, in one other (10%) there was an important decrease in the size of the cyst, and in one (10%), no change was seen. None of the patients had complications during the follow-up period of between 6 and 26 months (average 15.7 months). The authors postulate that a regimen of albendazole is the treatment of choice for this type of neurocysticercosis, although praziquantel may also be useful. PMID- 9202262 TI - Validation of the optic nerve sheath response to changing cerebrospinal fluid pressure: ultrasound findings during intrathecal infusion tests. AB - Raised intracranial pressure leads to increased pressure around the optic nerve (ON), which underlies the formation of papilledema and the enlargement of the dural optic nerve sheath (ONS). In clinical practice, the presence of widened ONSs is demonstrable on neuroimaging, but their relationship to cerebrospinal fluid (CSF) pressure remains unknown. The authors investigated the ONS response to pressure during CSF absorption studies in 12 patients undergoing neurological testing. The ONS diameter was evaluated by serial B-mode ultrasound scans of the anterior ON near its entry into the globe. All patients tested showed ONS diameter changes that exhibited covariance with the alteration of lumbar CSF pressure and were completely reversible during the infusion tests. The maximum difference in ONS diameter between baseline and peak pressure conditions was 1.8 mm on average (range 0.7-3.1 mm), corresponding to an average ONS diameter variation of 45% (range 15-89%). Regression analysis yielded a linear covariance between ONS diameter and CSF pressure with different slopes across subjects (0.019-0.071 mm/mm Hg, mean r = 0.78). However, this linear relationship was only present within a CSF pressure interval. This interval differed between patients: ONS dilation commenced at pressure thresholds between 15 mm Hg and 30 mm Hg and in some patients saturation of the response (constant ONS diameter) occurred between 30 mm Hg and 40 mm Hg. With a single exception, definitely enlarged ONS diameters (> 5 mm) were present when CSF pressure exceeded levels of 30 mm Hg. Retrospectively, discrimination between normal and elevated outflow resistance was possible on the basis of the ONS response to intrathecal infusion alone. It is concluded that the human ONS has sufficient elasticity to allow a detectable dilation in response to intracranial hypertension. Because of a variable pressure diameter relationship, the subarachnoid pressure cannot be predicted exactly by single scans. Therefore, the clinical relevance of this method relies on the demonstration of pathologically enlarged sheaths or ongoing enlargement on serial ultrasonography studies. PMID- 9202263 TI - Outpatient surgical treatment of cervical radiculopathy. AB - A series of 200 patients who underwent outpatient surgical treatment for cervical radiculopathy is presented. The patients were selected on the basis of their willingness to undergo surgery in the outpatient setting and the absence of serious underlying medical conditions. All operations were performed using general anesthetic techniques with limited posterior dissections. A laminoforaminotomy was performed at each affected level, which had been determined by preoperative imaging and clinical examination. After being observed for several hours, the patients were discharged if they met specific criteria. No patient required subsequent hospital admission in the immediate postoperative period. Follow-up review in 183 patients ranged from 3 to 43 months, with a mean of 19 months. In cases in which Workers' Compensation claims were not involved, 92.8% of patients reported an excellent or good outcome and returned to work or comparable duties at a mean of 2.9 weeks. In cases in which Workers' Compensation claims were involved, 77.8% of patients reported excellent or good outcome and returned to work at a mean of 7.6 weeks postoperatively. Two patients whose cases involved Workers' Compensation claims did not return to work. There were seven patients (3.8%) who had a poor outcome. Two of these patients underwent a second posterior procedure and reported a good outcome at the time of follow-up review. The results of this study show that outpatient surgical treatment of cervical radiculopathy can be safely provided in selected patients with outcomes similar to the inpatient surgical management of these individuals. PMID- 9202264 TI - Endoscopic endonasal transsphenoidal surgery: experience with 50 patients. AB - An endoscope was used in transsphenoidal surgery and eventually replaced the operating microscope as the tool for visualization. This study focuses on 50 patients (28 females and 22 males) with a median age of 38 years (range 14-88 years). Initially, four patients underwent operation via a sublabial-transseptal approach using a rigid endoscope in conjunction with an operating microscope. The 48 subsequent operations were performed through a nostril using only rigid endoscopes. Forty-four patients had pituitary adenomas and six had various other lesions. Thirteen patients had microadenomas, 16 had intrasellar macroadenomas, nine had macroadenomas with suprasellar extension, and six had invasive macroadenomas involving the cavernous sinus. Seven patients had recurrent pituitary adenomas and 25 had hormone-secreting adenomas (eight patients with Cushing's disease and 17 patients with prolactinomas). Among the eight patients with Cushing's disease, seven had resolution of hypercortisolism clinically and chemically. Of the 17 patients with prolactinomas, 10 improved clinically with normal serum prolactin levels, four improved clinically with elevated serum prolactin levels, and three had residual tumors in the cavernous sinus. Among the 19 patients with nonsecreting adenomas, 16 underwent total resection and three subtotal resection leaving residual tumor in the cavernous sinus. Postoperatively, all patients who had undergone endonasal endoscopic surgery had unobstructed nasal airways with minimal discomfort. More than half of the patients required only an overnight hospitalization. PMID- 9202265 TI - Microelectrode-guided posteroventral medial radiofrequency pallidotomy for Parkinson's disease. AB - The outcome of radiofrequency-guided posteroventral medial pallidotomy was investigated in 29 patients with recalcitrant Parkinson's disease. Extracellular recordings were obtained in the target region to differentiate the internal from the external globus pallidus, and distinct waveforms were recorded in each region. Stimulation of the target site further verified the lesion location. Of the 29 patients treated during the course of 1 year, none showed any adverse side effects (such as hemianopsia or hemiparesis) from the procedure. Significant and immediate improvement in motor involvement (dyskinesia, rigidity, dystonia, freezing, and tremor) was observed as measured by the Unified Parkinson's Disease Rating Scale and the Hoehn and Yahr scale. Patients experienced improvements in their condition as measured on a self-rating scale, and their ability to perform the activities of daily living was also significantly improved. Although the onset and duration of the effect of a single dose of levodopa did not change, the number of hours in an "off" state of dyskinesia per day was significantly decreased. These results provide further evidence, in a large group of patients, that posteroventral medial pallidotomy results in significant control of the motor symptoms of Parkinson's disease with a minimum of undesirable side effects. PMID- 9202266 TI - Facial nerve injury in acoustic neuroma (vestibular schwannoma) surgery: etiology and prevention. AB - Facial nerve injury associated with acoustic neuroma surgery has declined in incidence but remains a clinical concern. A retrospective analysis of 611 patients surgically treated for acoustic neuroma between 1973 and 1994 was undertaken to understand patterns of facial nerve injury more clearly and to identify factors that influence facial nerve outcome. Anatomical preservation of the facial nerve was achieved in 596 patients (97.5%). In the immediate postoperative period, 62.1% of patients displayed normal or near-normal facial nerve function (House-Brackmann Grade 1 or 2). This number rose to 85.3% of patients at 6 months after surgery and by 1 year, 89.7% of patients who had undergone acoustic neuroma surgery demonstrated normal or near-normal facial nerve function. The surgical approach appeared to have no effect on the incidence of facial nerve injury. Poor facial nerve outcome (House-Brackmann Grade 5 or 6) was seen in 1.58% of patients treated via the suboccipital approach and in 2.6% of patients treated via the translabyrinthine approach. When facial nerve outcome was examined with respect to tumor size, there clearly was an increased incidence of facial nerve palsy seen in the immediate postoperative period in cases of larger tumors: 60.8% of patients with tumors smaller than 2.5 cm had normal facial nerve function, whereas only 37.5% of patients with tumors larger than 4 cm had normal function. This difference was less pronounced, however, 6 months after surgery, when 92.1% of patients with tumors smaller than 2.5 cm had normal or near normal facial function, versus 75% of patients with tumors larger than 4 cm. The etiology of facial nerve injury is discussed with emphasis on the pathophysiology of facial nerve palsy. In addition, on the basis of the authors' experience with these complex tumors, techniques of preventing facial nerve injury are discussed. PMID- 9202267 TI - A new concept in Dorello's canal microanatomy: the petroclival venous confluence. AB - The so-called Dorello's canal was studied in 32 specimens (16 human cadaver heads) injected with colored latex and fixed in formalin (28 specimens) or studied with microscopic and ultrastructural methods (four specimens). To avoid the differences usually encountered in the description of this area, the authors preferred to consider a larger space that they have named the petroclival venous confluence (PVC). It was located between two dural layers: inner (or cerebral) and outer (or osteoperiosteal). The PVC was quadrangular on transverse section. The posterior petroclinoid fold and the axial plane below the dural foramen of the abducent nerve (sixth cranial nerve) limited the PVC at the top and bottom, respectively. Its anteroinferior limit was the posterosuperior aspect of the upper clivus and outer layer of the dura mater. Its anterior limit was the vertical plane containing the posterior petroclinoid fold, and its posterior limit was the inner layer of the dura. The PVC was limited laterally by the medial aspect of the petrous bone apex and medially by the virtual sagittal plane extending the medial limit of the inferior petrosal sinus upward. The PVC was a venous space bordered by endothelium and continuous with the cavernous sinus, the basal sinus of the clivus, and the inferior petrosal sinus. There were trabeculations between the two dural layers. The petrosphenoidal ligament of Gruber may be regarded as a larger trabeculation, and it divided the PVC into a superior and an inferior compartment. The abducent nerve generally ran through the inferior compartment, where it was fixed to the surrounding dura mater. This nerve was only separated from venous blood by a meningeal sheath of varying thinness lined with endothelium. The clinical implications of these findings are discussed. PMID- 9202268 TI - Seizures induced by intracerebral injection of thrombin: a model of intracerebral hemorrhage. AB - The coagulation cascade plays an important role in brain edema formation caused by intracerebral blood. In particular, thrombin produces brain injury via direct brain cell toxicity. Seizures and increased cerebral electrical activity are commonly associated with intracerebral blood and are possible effects of thrombin leading to cell injury in the brain. In this study, artificial clots containing concentrations of thrombin found in hematomas were infused intracerebrally in rats. The animals were observed clinically for seizure activity, behavior, and neurological deficits. Several animals underwent video electroencephalographic (EEG) monitoring during intracerebral infusion and for 30 minutes postinfusion. All animals were killed 24 hours after injection, and brain water and ion contents were measured to determine the amount of brain edema. Clinically, thrombin produced focal motor seizures in all animals. None of the control animals or those receiving N[alpha]-(2-Naphthalenesulfonyl-glycyl)-4-amidino-DL phenylalanine -piperidide (alpha-NAPAP), a thrombin inhibitor added to the thrombin, showed clinical evidence of seizures. Of the rats undergoing EEG monitoring, all animals receiving thrombin showed electrical evidence of seizure activity, whereas none of the control animals exhibited seizure activity. There was no evidence of seizure activity on EEG monitoring when alpha-NAPAP was injected along with the thrombin. In addition, the artificial clots containing thrombin produced agitation and a circling tendency in the rats, along with brain edema. These results indicate that the coagulation cascade is involved in seizure production and increased brain electrical activity, which contribute to the neurological deficits and brain edema formation that are seen with intracerebral hemorrhage. PMID- 9202269 TI - Selective motor hyperreinnervation using motor rootlet transfer: an experimental study in rat brachial plexus. AB - Misdirection of sensory fibers into motor pathways is, in part, responsible for the poor results obtained after peripheral nerve repair. After avulsion of the C 5 root in rats, the authors connected a C-4 ventral rootlet to the musculocutaneous nerve by means of a sural nerve graft. In this way, they were able to increase the number of regenerating motor fibers and avoid growth of sensory fibers into the nerve grafts. Functional recovery was evaluated electrophysiologically and histologically. The origin of the axons that reinnervated the nerve graft was analyzed by means of morphological studies including retrograde labeling procedures. Motor neurons survived and regenerated after the rootlet transfer and there was no functional impairment. Many neurons were retrograde labeled in the ventral horn and widespread biceps muscle reinnervation was demonstrated with recovery of nearly normal electrophysiological properties. Motor hyperreinnervation of the musculocutaneous nerve was observed. This high degree of reinnervation in a long (40-mm) graft was attributed to the good chance that a muscle fiber can be reinnervated by a motor fiber when the number of regenerating motor neurons is increased and when competitive sensory fibers are excluded from reinnervation. PMID- 9202270 TI - A comparison of proliferation indices in human anterior pituitary adenomas using formalin-fixed tissue and in vitro cell culture. AB - The authors compared detection methods for cell proliferation in human anterior pituitary adenomas using histological sections and dispersed cell culture. After tumor cells had been grown for 4 days in dispersed culture, bromodeoxyuridine (BUdR), proliferating cell nuclear antigen (PCNA), and Ki-67 were compared by double immunostaining and contrasted with single staining of PCNA and Ki-67 indices in the corresponding histological sections from 12 human pituitary adenomas. In vitro, the BUdR labeling index was positive in six of 12 tumors (range < 0.1-5.1%), 10 of 12 tumors were PCNA-positive (range < 0.1-100%), and Ki 67 was positive in 10 of 12 adenomas (range < 0.1-8%). In vitro, BUdR and Ki-67 gave similar proliferative indices for 10 of 12 adenomas. In vivo, the PCNA labeling index was positive in 12 of 12 adenomas (range 0.9-95%) and Ki-67 was positive in 11 of 12 adenomas (range < 0.1-2%). Tumors with a labeling index less than 0.1% were considered to be negative for proliferation. High PCNA values were found in vitro and in vivo, whereas Ki-67 labeling indices were similar in vitro and in vivo for nine of 12 adenomas. It is concluded that Ki-67 proliferative indices in vivo reflect those found in vitro, at least after 4 days in dispersed culture, but that PCNA overestimates pituitary adenoma proliferation in histological sections as well as in dispersed culture. PMID- 9202271 TI - Regulated expression of the diphtheria toxin A gene in human glioma cells using prokaryotic transcriptional control elements. AB - Because accurate regulation of toxin gene expression is critical for safe and effective gene therapy applications, the authors have examined the regulation of diphtheria toxin A (DTA) fragment expression in human glioma cell lines using two transcriptional control systems derived from Escherichia coli: the tetracycline (Tet) system and the lactose (Lac) system. The Tet system includes a tetracycline controlled transactivator (tTA), a tTA-responsive minimum human cytomegalovirus (hCMV) promoter controlling the expression of the DTA gene, and tetracycline as an allosteric inhibitor. The Lac system includes the lac repressor (lacR), a lacR regulated Rous sarcoma virus-long terminal repeat (RSV-LTR) promoter controlling the expression of the DTA gene, and isopropyl-thio-beta-D-galactoside (IPTG) as an allosteric inducer. Expression plasmids encoding either tTA or lacR were transfected into U-87MG and U-343MG glioma cells along with the responsive DTA plasmid. Cell killing was monitored by the ability of the toxin to abolish protein synthesis and was quantitated using a luciferase reporter gene. In the Tet system, tumor cell killing could be regulated by tetracycline up to 120-fold. In contrast, only a twofold IPTG-dependent regulation was obtained using the Lac system because of an incomplete repression of DTA expression in the uninduced state. Replacement of the RSV-LTR promoter with the heavy metal-inducible mouse metallothionein-1 promoter in the lacR-responsive unit, as well as the generation of a clonal glioma cell line expressing lacR, did not significantly enhance regulation of DTA in the Lac system. In conclusion, this study demonstrates that the Tet system is of potential use in gene therapy applications in which regulated expression of a therapeutic gene is an important issue. PMID- 9202272 TI - Triple anterior screw fixation of an acute combination atlas-axis fracture. Case report. AB - The authors report the successful treatment of an acute combination atlas-axis fracture in an 85-year-old man using anterior odontoid and C1-2 transarticular facet screw fixation and a Philadelphia collar. Treatment with halo brace immobilization failed, and the patient experienced recurrent episodes of oxygen desaturation when placed partially prone for chest physiotherapy. If a posterior approach is not feasible, an anterior odontoid and C1-2 transarticular facet screw fixation can be considered as a salvage procedure for patients with acute combination atlas-axis fractures. PMID- 9202273 TI - Neurocytoma in the cerebellum. Case report. AB - A neurocytoma is a central nervous system tumor composed of small cells with features of neuronal differentiation; it typically occurs in the periventricular region, close to the septum pellucidum and the foramen of Monro. In this article, the authors report the case of a neurocytoma located in the cerebellum, which to their knowledge is the first reported case of its kind. The finding of a neurocytoma at a nonclassic location refutes the theory that this tumor has its origins in subependymal progenitor cells, unless an ectopic location of progenitor cells is invoked to explain the occurrence of a neurocytoma away from the ventricles. On the basis of this case, the authors suggest that neurocytomas should be added to the differential diagnosis of mass lesions in the supratentorial intraventricular regions as well as in the posterior fossa. PMID- 9202274 TI - Third ventricular choroid plexus papilloma with psychosis. Case report. AB - This 9-year-old boy with a history of behavioral problems and worsening psychosis responded initially to treatment with haloperidol. However, a magnetic resonance image obtained as part of his psychiatric evaluation revealed an anterior third ventricle tumor and mild-to-moderate hydrocephalus. The resected tumor was found on pathological examination to be a choroid plexus papilloma. The patient had an uneventful postoperative course and remained free of psychosis or mood disorder at 1-year follow-up examination. PMID- 9202275 TI - Stenotrophomonas maltophilia meningitis. Report of two cases and review of the literature. AB - The authors report two cases of meningitis caused by Stenotrophomonas maltophilia in cancer patients following placement of an Ommaya reservoir for treatment of meningeal carcinomatosis. In addition, they review eight other cases of S. maltophilia that have been reported to date. Stenotrophomonas maltophilia meningitis is often associated with neurosurgical procedures; however, spontaneous infection may also occur, mainly in neonates. The disease's clinical presentation is similar to that of other forms of meningitis caused by Gram negative bacilli. The overall mortality rate of this disease is 20% and is limited to neonates with spontaneous meningitis in whom effective antibiotic therapy is delayed. Meningitis caused by S. maltophilia in the modern era should be considered in immunocompromised hosts with significant central nervous system disease who have undergone neurosurgical procedures and who do not readily respond to broad-spectrum antimicrobial coverage. PMID- 9202276 TI - Cerebral abscess as an unusual complication of coil embolization in a dural arteriovenous fistula. Case report. AB - This 63-year-old man presented with a right temporoparietal cortical infarction. A dural arteriovenous fistula involving the right transverse sinus was diagnosed on cerebral angiography. Transvenous embolization using detachable coils was performed; however, postembolization angiograms demonstrated retrograde filling of a cortical draining vein that was not seen on initial angiography. The patient subsequently developed a cerebral abscess in the region of the previous cortical infarction 2 months after the embolization. The abscess was successfully treated with drainage and antibiotic therapy. The authors report this case to illustrate an unusual complication associated with this procedure and the possible contribution of the cortical draining vein in the pathogenesis of the cerebral abscess. PMID- 9202277 TI - Function and organization in dysgenic cortex. Case report. AB - Cerebral dysgenesis is a subject of interest because of its relationship to cerebral development and dysfunction and to epilepsy. The authors present a detailed study of a 16-year-old boy who underwent surgery for a severe seizure disorder. This patient had dysgenesis of the right hemisphere, which was composed of a giant central frontoparietal nodular gray matter heterotopia with overlying large islands of cortical dysplasia around a displaced central fissure. Exceptional insight into the function, biochemistry, electrophysiology, and histological structure of this lesion was obtained from neurological studies that revealed complementary information: magnetic resonance (MR) imaging, [18]fluoro-2 deoxy-D-glucose positron emission tomography (PET), functional PET scanning, proton MR spectroscopic (1H-MRS) imaging, intraoperative cortical mapping and electrocorticography, in vitro electrophysiology, and immunocytochemistry. These studies demonstrated compensatory cortical reorganization and showed that large areas of heterotopia and cortical dysplasia in the central area may retain normal motor and sensory function despite strikingly altered cytoarchitectonic organization and neuronal metabolism. Such lesions necessitate appropriate functional imaging studies prior to surgery and cortical mapping to avoid creating neurological deficits. Integrated studies, such as PET, 1H-MRS imaging, cortical mapping, immunocytochemistry, and electrophysiology may provide information on the function of developmental disorders of cerebral organization. PMID- 9202278 TI - Diagnostic biopsy of the motor nerve to the gracilis muscle. Technical note. AB - A biopsy specimen of the motor nerve to the gracilis muscle was obtained in 151 patients to aid in the diagnosis of neuromuscular disorders. The procedure is simple, safe, and has low morbidity and complication rates. The surgical technique is described. Indications for the use of this technique are motor neuropathy, motor neuron disease, Guillain-Barre syndrome, and mixed neuropathy with prominent motor symptoms. PMID- 9202279 TI - Endoscopic microscopic transpedicular thoracic discectomy. Technical note. AB - In an effort to make thoracic discectomy simple and less invasive while using direct visualization, a 70 degrees-angled lens endoscope has been adopted to visualize the ventral aspect of the spinal cord dura mater during microsurgical thoracic discectomy via a transpedicular approach. The patient is positioned in a 60 degrees forwardly inclined lateral position with the side of the lesion facing upward. After radiographic corroboration of the correct level, a transpedicular approach is made using a 1.5-cm-diameter tubular retractor through a 2-cm-long paramedian transverse skin incision. With the aid of an operating microscope, the ipsilateral facet joint, including the upper portion of the pedicle, is removed using a high-speed drill, thus exposing the neural foramen, intervertebral disc, and upper portion of the pedicle leading to the vertebral bodies. When the herniated disc and bone spur have been removed laterally in relation to the spinal cord, creating a cavity under the operating microscope, a 4-mm-diameter rigid endoscope with a 70 degrees-angled lens is mounted to an endoscope holder so that the ventral aspect of the spinal cord dura mater can be visualized directly. With the aid of direct endoscopic visualization, the disc and bone spur, which compress the spinal cord anteriorly, are pushed away toward a cavity created at the intervertebral space and are removed using a downward-biting long armed curette. Patients with myelopathy are kept overnight in the hospital; however, those with radiculopathy are discharged home on the same day as their operation. The surgical technique and two illustrative cases are reported. PMID- 9202280 TI - Venous varix causing median neuropathy. Case illustration. PMID- 9202281 TI - Pituitary astrocytoma: magnetic resonance and hormonal characteristics. Case illustration. PMID- 9202282 TI - Extradural hematoma following maxillary sinusitis. Case illustration. PMID- 9202283 TI - Hearing and vestibular schwannomas. PMID- 9202284 TI - Astrocytoma and Ki-67. PMID- 9202285 TI - Brain tissue pressure gradients. PMID- 9202286 TI - Posttraumatic vasospasm. PMID- 9202287 TI - Pott's disease. PMID- 9202288 TI - Novel isoforms of rat brain fructose 6-phosphate 2-kinase/fructose 2,6 bisphosphatase are generated by tissue-specific alternative splicing. AB - Fructose 6-phosphate 2-kinase/fructose 2,6-bisphosphatase catalyzes the synthesis and degradation of fructose 2,6-bisphosphate, which is the most potent activator of glycolysis. We have shown previously the occurrence of a partial cDNA (RB7) encoding the catalytic core domain of a novel brain-type isozyme. To elucidate the full-length sequence of RB7 cDNA, we have carried out cDNA cloning using the 3'- and 5'-rapid amplification cDNA ends method and have isolated eight isoforms from rat brain. The cDNA sequences encoding the 5'-untranslated region, the amino terminal domain, and the catalytic core domain were identical among all the isoforms. However, heterogeneity of the carboxyl-terminus was found by sequence analysis. This heterogeneity was shown not to have resulted from transcription of multiple genes, as Southern blot and genomic sequence analysis revealed that the gene was a single copy in the rat genome. It is likely that these transcripts represent splice variants of the gene. High-level expression of the gene was also observed by northern blot analysis in skeletal muscle. However, the pattern of alternative splicing was different from that of brain, and only four isoforms were detected in skeletal muscle. PMID- 9202289 TI - ICE/CED-3 family executes oligodendrocyte apoptosis by tumor necrosis factor. AB - Tumor necrosis factor (TNF) is thought to be one of the mediators responsible for the damage of oligodendrocytes (OLGs) in multiple sclerosis (MS). We report here the involvement of the interleukin 1beta-converting enzyme (ICE)/Caenorhabditis elegans gene ced-3 (CED-3) family in TNF-mediated cell death of OLGs. The addition of TNF-alpha to primary cultures of OLGs that express ice and cpp32 significantly decreased the number of live OLGs in 72 h. DNA fragmentation was detected in TNF-treated OLGs at 36 h with the terminal deoxynucleotidyl transferase dUTP nick end-labeling assay. Benzyloxycarbonyl-Asp-CH2OC(O)-2,6 dichlorobenzene, an inhibitor of the ICE/CED-3 family that shows p35-like inhibitory specificity, protected against the TNF-induced cell death of OLGs. Furthermore, acetyl-YVAD-CHO (a specific inhibitor of ICE-like proteases) as well as acetyl-DEVD-CHO (a specific inhibitor of CPP32-like proteases) enhanced the survival of OLGs treated with TNF-alpha, indicating that ICE- and the CPP32 mediated cell death pathways are activated in TNF-induced OLG cell death. Our results suggest that the inhibition of ICE/CED-3 proteases may be a novel approach to treat neurodegenerative diseases such as MS. PMID- 9202291 TI - Distribution of brain-derived neurotrophic factor in rats and its changes with development in the brain. AB - A newly established, sensitive, two-site enzyme-immunoassay system for brain derived neurotrophic factor (BDNF) is described. Using this system, we investigated the tissue distribution of BDNF and developmental changes in tissue levels of BDNF in rats. The minimal limit of detection of the assay was 3 pg/0.2 ml of assay mixture. BDNF was successfully solubilized from tissues in the presence of guanidine hydrochloride but not in any of the other buffers examined. In the rat brain at 1 month of age, the highest level of BDNF was detected in the hippocampus (5.41 ng/g of wet weight), followed by the hypothalamus (4.23 ng/g) and the septum (1.68 ng/g). In other regions, levels of BDNF ranged between 0.9 and 1.7 ng/g. The level of BDNF in the posterior lobes of the cerebellum from rats at 30 days of age was slightly higher than that in the anterior lobes. The concentration of BDNF increased in all regions of the brain with postnatal development. In peripheral tissues, BDNF was found at very low concentrations (0.65 ng/g in the spleen, 0.21 ng/g in the thymus, and 0.06 ng/g in the liver). The subfractionation of the hippocampal homogenate indicated that approximately 50% of BDNF was contained in the crude nuclear fraction. Immunoblots of BDNF immunoreactive proteins extracted from the hippocampus, hypothalamus, and cerebellum contained doublet bands of protein of approximately 14 kDa, a value close to the molecular mass of recombinant human BDNF. Immunocytochemical investigations showed that, in the hippocampus, BDNF was localized in the nucleus of the granule cells in the dentate gyrus and of the cells in the pyramidal cell layer. The frequency of cells that were stained in the dentate gyrus was greater than that of cells in the pyramidal cell layer. PMID- 9202290 TI - Effects of wild-type and mutated copper/zinc superoxide dismutase on neuronal survival and L-DOPA-induced toxicity in postnatal midbrain culture. AB - Mutations in the free radical-scavenging enzyme copper/zinc superoxide dismutase (Cu/Zn-SOD) are associated with neuronal death in humans and mice. Here, we examine the effects of human wild-type (WT SOD) and mutant (Gly93 --> Ala; G93A) Cu/Zn-SOD enzyme on the fate of postnatal midbrain neurons. One-week-old cultures from transgenic mice expressing WT SOD enzyme had significantly more midbrain neurons and fewer necrotic and apoptotic neurons than nontransgenic cultures. In contrast, 1-week-old cultures from transgenic G93A mice expressing mutant SOD enzyme had significantly fewer midbrain neurons and more necrotic and apoptotic neurons than nontransgenic cultures. To subject postnatal midbrain neurons to oxidative stress, cultures were incubated with L-DOPA. L-DOPA at 200 microM caused approximately 50% loss of tyrosine hydroxylase (TH)-positive neurons in nontransgenic cultures and even greater loss in transgenic G93A cultures; no alterations were noted in GABA neuron numbers. In contrast, 200 microM L-DOPA did not cause any significant reductions in TH-positive or GABA neuron numbers in transgenic WT SOD cultures. L-DOPA at 50 microM had opposite effects, in that it significantly increased TH-positive, but not GABA neuron numbers in transgenic WT SOD and G93A and in nontransgenic cultures. These results indicate that increased amounts of WT SOD enzyme promote cell survival and protect against L-DOPA-induced dopaminergic neurotoxicity, whereas increased amounts of mutated Cu/Zn-SOD enzyme have inverse effects. As the spontaneous loss and L-DOPA-induced loss of postnatal dopaminergic midbrain neurons appear to be mediated by free radicals, our study supports the view that mutated Cu/Zn-SOD enzyme kills cells by oxidative stress. PMID- 9202292 TI - Dramatic increase of the RNA editing for glutamate receptor subunits during terminal differentiation of clonal human neurons. AB - RNA editing plays an important role in determining physiological characteristics of certain glutamate-gated receptor (GluR) channels such as Ca2+ permeability and desensitization kinetics. In one case, the editing changes a gene-encoded glutamine (Q) to an arginine (R) codon located in the channel-forming domain of the alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor subunit GluR-B and also the kainate receptor subunits GluR5 and GluR6. Another case of RNA editing alters an arginine (R) to a glycine (G) codon at a position termed the "R/G" site of AMPA subunits GluR-B, C, and D. Double-stranded RNA-specific adenosine deaminases (DRADA) have been implicated as agents involved in the editing. By using a human teratocarcinoma cell line, NT2, we investigated the change of the RNA editing of GluR subunits in conjunction with the expression of two DRADA members, DRADA1 and DRADA2 genes, during neuronal differentiation. Whereas Q/R and R/G site RNA editing both become progressively activated in differentiating NT2 cells, the expression of the two DRADA genes can already be detected even in the undifferentiated NT2 cells. Development of the editing machinery appears to require, in addition to DRADA enzymes, a currently unidentified mechanism(s) that may become activated during neuronal differentiation. PMID- 9202293 TI - Nerve growth factor protects PC12 cells against peroxynitrite-induced apoptosis via a mechanism dependent on phosphatidylinositol 3-kinase. AB - Nerve growth factor (NGF) prevents apoptosis induced by the oxidant peroxynitrite in undifferentiated PC12 rat pheochromocytoma cells. Previous studies have shown that activation of phosphatidylinositol 3-kinase (PI 3-kinase) by NGF via the TrkA receptor tyrosine kinase protects PC12 cells from serum deprivation-induced apoptosis. We found that two PI 3-kinase inhibitors, wortmannin and LY294002, eliminated the protection NGF provided against peroxynitrite-induced apoptosis at concentrations consistent with their effectiveness as PI 3-kinase inhibitors. When the activity of PI 3-kinase was assayed in phosphotyrosine immunoprecipitates after treatment of PC12 cells with peroxynitrite, PI 3-kinase activity was reduced by 50% of that detected in control cells, whereas PI 3 kinase activity in NGF-treated cells was unaffected by peroxynitrite. If an antibody against PI 3-kinase was used to immunoprecipitate the enzyme, treatment with peroxynitrite had no effect on activity. Therefore, peroxynitrite appeared to disrupt interactions between PI 3-kinase and phosphotyrosine proteins, rather than directly inhibiting the enzyme. NGF also activates p21Ras-dependent pathways, but this did not appear to be required for NGF to exert its protective effect against peroxynitrite. PC12 cells expressing a dominant inhibitory mutant of p21Ras were equally susceptible to peroxynitrite-induced apoptosis, which was prevented by NGF. Wortmannin was also able to block the protective effect of NGF in the p21Ras mutant cell line. Although many signaling pathways are activated by NGF, these results suggest that a PI 3-kinase-dependent pathway is important for inhibiting peroxynitrite-induced apoptosis. PMID- 9202294 TI - Isoform-specific modulation by apolipoprotein E of the activities of secreted beta-amyloid precursor protein. AB - The genes for both the beta-amyloid precursor protein and apolipoprotein E (ApoE) have been linked to Alzheimer's disease. This connection suggests the possibility that these proteins interact physically or functionally. To explore this idea, we focused on the neuroprotective activity of secreted amyloid precursor protein (sAPP) and related signal transduction events. After coincubation with ApoE, sAPP exhibited an enhanced [Ca2+]i-lowering activity and enhanced protection against excitotoxicity in rat primary hippocampal neurons. In contrast, the stimulation of phosphoinositide production by sAPP was inhibited by ApoE. Kinetic analyses and coimmunoprecipitation experiments indicated that these actions result from formation of a heteromeric complex between ApoE and sAPP. Furthermore, the ApoE4 isoform, which seems to accelerate the onset of Alzheimer's disease, was less potent than ApoE3 in modifying each activity of sAPP. These data suggest that sAPP-dependent neuroprotective mechanisms would be compromised in individuals expressing ApoE4, a scenario that may contribute to the development of Alzheimer's disease. PMID- 9202295 TI - Lysophosphatidic acid induces a sustained elevation of neuronal intracellular calcium. AB - Lysophosphatidic acid (LPA) is a lipid biomediator enriched in the brain. A novel LPA-induced response in rat hippocampal neurons is described herein, namely, a rapid and sustained elevation in the concentration of free intracellular calcium ([Ca2+]i). This increase is specific, in that the related lipids phosphatidic acid and lysophosphatidylcholine did not induce an alteration in [Ca2+]i. Moreover, consistent with a receptor-mediated process, there was no further increase in [Ca2+]i after a second addition of LPA. The LPA-induced increase in [Ca2+]i required extracellular calcium. However, studies with Cd2+, Ni2+, and nifedipine and nystatin-perforated patch clamp analyses did not indicate involvement of voltage-gated calcium channels in the LPA-induced response. In contrast, glutamate appears to have a significant role in the LPA-induced increase in [Ca2+]i, because this increase was inhibited by NMDA receptor antagonists and alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA)/kainate receptor antagonists. Thus, LPA treatment may result in an increased extracellular glutamate concentration that could stimulate AMPA/kainate receptors and thereby alleviate the Mg2+ block of the NMDA receptors and lead to glutamate stimulation of an influx of calcium via NMDA receptors. PMID- 9202296 TI - Basic fibroblast growth factor stimulation of glial cells protects dopamine neurons from 6-hydroxydopamine toxicity: involvement of the glutathione system. AB - Neurotrophic factors have been shown to support the survival and promote the recovery of injured neurons both in vivo and in vitro. Here, we investigated whether glial cell line-derived neurotrophic factor (GDNF) and basic fibroblast growth factor (bFGF) could modify the damage to dopamine (DA) neurons in mesencephalic cultures caused by the neurotoxin 6-hydroxydopamine (6-OHDA). The data show that bFGF, but not GDNF, effectively protected DA neurons from 6-OHDA toxicity. Because bFGF is a glial mitogen, whereas GDNF is not, we tested whether glial cells participated in bFGF neuroprotection. Inhibition of glial cell proliferation completely prevented the protective effect of bFGF. Because oxidative events have been associated with 6-OHDA-induced damage, we examined the levels of glutathione (GSH) in control and bFGF-treated cultures. Cultures treated with bFGF had higher levels of GSH, which increased even further in response to 6-OHDA exposure. Control cultures failed to up-regulate GSH levels after 6-OHDA, suggesting a relationship between increased GSH levels and protection from 6-OHDA. Inhibition of glial cell proliferation prevented the rise in GSH in bFGF-treated cultures and abolished the increase after 6-OHDA treatment. Protection from 6-OHDA by bFGF was also diminished when GSH levels were decreased by the GSH synthesis inhibitor L-buthionine sulfoximine. Our study shows that stimulation of glial cells by bFGF allows the up-regulation of antioxidant defenses and supports cell survival during oxidative stress. PMID- 9202297 TI - Expression of the Na+-D-glucose cotransporter SGLT1 in neurons. AB - In brains of the rabbit, pig, and human, expression of the high-affinity Na+-D glucose cotransporter SGLT1 and of the protein RS1, which alters the activity of SGLT1, was demonstrated. In situ hybridization showed that SGLT1 and RS1 are transcribed in pyramidal cells of brain cortex and hippocampus and in Purkinje cells of cerebellum. In neurons of pig brain SGLT1 protein was demonstrated by western blotting with synaptosomal membranes and by immunohistochemistry, which showed SGLT1 in pyramidal and Purkinje cells. To test whether SGLT1 in neurons may be activated during increased D-glucose consumption, an epileptic seizure was induced in rat brain, and the uptake of specific nonmetabolized substrates of SGLT1 [[14C]methyl-alpha-D-glucopyranoside ([14C]AMG)] and of Na+-independent transporters [2-deoxy-D-[14C] glucose ([14C]2-DG)] was analyzed by autoradiography. During the seizure the uptake of AMG and 2-DG was increased in the focus. Within two hours after the seizure 2-DG uptake in the focus returned to normal. In contrast, the AMG uptake in the focus area was still increased 1 day later. The data show that the high-affinity Na+-D-glucose cotransporter SGLT1 is expressed in neurons and can be up-regulated. PMID- 9202298 TI - Construction and characterization of ciliary neurotrophic factor (CNTF) antagonists: microenvironmental difference in the CNTF receptor between rat and chicken cells for recognizing the D1 cap region. AB - Antagonistic mutants of ciliary neurotrophic factor (CNTF) were constructed and their properties characterized. K155A and K155W mutants lost cell survival promoting activity for chicken dorsal root ganglion (DRG) neurons and inhibited the activity of the wild type. However, they retained slight agonistic activity for the survival of rat DRG neurons, indicating there is a difference between chicken and rat cells for receptor recognition around the D1 cap region including K155 residue. The chicken receptor recognizes the D1 cap region more strictly than does the rat receptor. The substitution of F152, which locates at the top of the D1 cap region, was combined with the K155A mutation. A combination of the two mutations gave an antagonistic feature to not only chicken but also rat cells. Both F152S/K155A and F152D/K155A mutants lacked cell survival promoting activity and had an antagonistic effect on rat DRG neurons. The three-dimensional structure of CNTF suggests the following. F152 and K155 bind to the receptor with hydrophobic and electrostatic interactions, respectively. F152 locates close to L156 with a van der Waals contact, and K155 contacts with Q42 through a hydrogen bond. Both interactions play indispensable roles in maintaining the structure around the D1 cap region of CNTF. PMID- 9202299 TI - Effect of calcitriol in combination with corticosterone, interleukin-1beta, and transforming growth factor-beta1 on nerve growth factor secretion in an astroglial cell line. AB - In astrocytes, nerve growth factor (NGF) synthesis has been described to be stimulated by the cytokines interleukin-1beta (IL-1beta) and transforming growth factor-beta1 (TGF-beta1) and inhibited by corticosterone. As all three factors are present in the brain under certain conditions, we investigated the effect of their combined application on NGF secretion in the astroglial cell line RC7 and, in addition, studied the effect of calcitriol (1alpha,25-dihydroxyvitamin D3). Calcitriol stimulated NGF secretion, whereas corticosterone reduced basal levels of NGF secretion as well as inhibited the NGF secretion induced by IL-1beta, calcitriol, and TGF-beta1. Calcitriol had an additive effect when applied together with IL-1beta and a synergistic effect when applied with TGF-beta1. Moreover, calcitriol not only counteracted the inhibitory effect of corticosterone on NGF secretion stimulated by TGF-beta1 but even augmented it to a level more than threefold higher than that reached with TGF-beta1 alone. Due to the trophic effect of NGF on basal forebrain cholinergic neurons, these findings might be of therapeutic relevance under conditions where cholinergic function is impaired and the endogenous levels of corticosterone, IL-1beta, or TGF-beta1 are elevated. PMID- 9202300 TI - Nitrite and nitrate levels in individual molluscan neurons: single-cell capillary electrophoresis analysis. AB - Cell and tissue concentrations of NO2- and NO3- are important indicators of nitric oxide synthase activity and crucial in the regulation of many metabolic functions, as well as in nonenzymatic nitric oxide release. We adapted the capillary electrophoresis technique to quantify NO2- and NO3- levels in single identified buccal neurons and ganglia in the opisthobranch mollusc Pleurobranchaea californica, a model system for the study of the chemistry of neuron function. Neurons were injected into a 75-microm separation capillary and the NO2- and NO3- were separated electrophoretically from other anions and detected by direct ultraviolet absorbance. The limits of detection for NO2- and NO3- were <200 fmol (<4 microM in the neurons under study). The NO2- and NO3- levels in individual neurons varied from 2 mM (NO2-) and 12 mM (NO3-) in neurons histochemically positive for NADPH-diaphorase activity down to undetectable levels in many NADPH-diaphorase-negative cells. These results affirm the correspondence of histochemical NADPH-diaphorase activity and nitric oxide synthase in molluscan neurons. NO2- was not detected in whole ganglion homogenates or in hemolymph, whereas hemolymph NO3- averaged 1.8 +/- 0.2 x 10(-3) M. Hemolymph NO3- in Pleurobranchaea was appreciably higher than values measured for the freshwater pulmonate Lymnaea stagnalis (3.2 +/- 0.2 x 10(-5) M) and for another opisthobranch, Aplysia californica (3.6 +/- 0.7 x 10(-4) M). Capillary electrophoresis methods provide utility and convenience for monitoring NO2-/NO3- levels in single cells and small amounts of tissue. PMID- 9202302 TI - Bidirectional alterations of GABA(A) receptor subunit peptide levels in rat cortex during chronic ethanol consumption and withdrawal. AB - The pharmacological properties of gamma-aminobutyric acidA (GABA(A)) receptors are altered by prolonged exposure to ethanol both in vivo and in vitro. We have shown previously that prolonged ethanol exposure elicits selective alterations in various GABA(A) receptor subunit mRNA levels in rat cerebral cortex. Some of these effects are rapidly reversed during ethanol withdrawal. The present study was conducted to determine the effects of prolonged ethanol exposure (dependence) and ethanol withdrawal on cerebral cortical peptide expression for several subunits. GABA(A) receptor alpha1 subunit peptide levels were decreased by nearly 40%, whereas alpha4 subunit peptide levels were increased by 27% in both ethanol dependent and withdrawn rats. These changes correlate well with observed alterations in mRNA levels following prolonged ethanol exposure in dependent rats, but do not match the effects on mRNA levels during ethanol withdrawal. Beta2/3 subunit peptide levels increased by approximately 32% in both ethanol dependent rats and rats undergoing ethanol withdrawal. We observed a 30-60% increase in gamma1 subunit peptide levels in both dependent rats and those undergoing withdrawal, also correlating with the previous report on ethanol induced alterations in mRNA levels. Peptide levels for gamma2 subunits did not differ from control values in either condition. These findings show that specific alterations in GABA(A) receptor subunit peptide levels are associated with ethanol dependence in rats. GABA(A) receptor subunit peptide expression is more stable than mRNA expression, and mRNA levels are not representative of peptide expression during ethanol withdrawal. These findings are consistent with the suggestion that alterations in GABA(A) receptor gene expression underlie the functional properties of GABA(A) receptors in ethanol-dependent rats and those undergoing ethanol withdrawal. PMID- 9202301 TI - Ganglioside GM1 activates the mitogen-activated protein kinase Erk2 and p70 S6 kinase in U-1242 MG human glioma cells. AB - Gangliosides are implicated in the regulation of cellular proliferation as evidenced by differences in ganglioside composition associated with malignant transformation and density of cells in culture, as well as their inhibitory effects when added to cells growing in culture. Exogenously added gangliosides have a bimodal effect on proliferation in U-1242 MG glioma cells, inhibiting DNA synthesis in growing cells and stimulating it in quiescent cells. We investigated the mechanisms involved in stimulation of DNA synthesis using [3H]thymidine incorporation and immune complex kinase assays to identify responsible signal transduction pathways. Treatment of quiescent U-1242 MG cells with GM1 caused activation of the mitogen-activated protein (MAP) kinase isoform Erk2. Pretreatment with the specific MAP kinase kinase inhibitor PD98059 prevented the GM1-stimulated Erk2 activation and GM1-stimulated DNA synthesis. GM1 treatment stimulated another distinct signaling pathway leading to activation of p70 S6 kinase (p70s6k), and this was prevented by pretreatment with rapamycin. Rapamycin also inhibited GM1-stimulated DNA synthesis. Activation of both pathways and stimulation of DNA synthesis were inhibited by forskolin treatment; however, GM1 had no effect on cyclic AMP levels. Platelet-derived growth factor also activated both Erk2 and p70s6k but did not cause DNA synthesis, suggesting that GM1 may stimulate additional cascades, which also contribute to GM1-mediated DNA synthesis. PMID- 9202303 TI - NMDA receptor-mediated regulation of AMPA receptor properties in organotypic hippocampal slice cultures. AB - Activation of the calcium-dependent protease calpain has been proposed to be a necessary step in the formation of long-term potentiation (LTP) in the hippocampus, and stimulation of N-methyl-D-aspartate (NMDA) receptors leads to an increase in intracellular calcium concentration, calpain activation, proteolysis of cytoskeletal elements, and modification of alpha-amino-3-hydroxy-5-methyl-4 isoxazole propionic acid (AMPA) receptor properties. In the present study, we evaluated the effects of NMDA treatment of cultured hippocampal slices on the properties of AMPA receptors. Cultured hippocampal slices were treated with NMDA (100 microM) for 15 min and [3H]AMPA binding to membrane fractions was measured. NMDA-treated slices exhibited an increase in both "high-affinity" and "low affinity" [3H]AMPA binding, with smaller changes in 6-cyano-7 nitro[3H]quinoxaline-2,3-dione binding. The increase in [3H]AMPA binding was significantly reduced by preincubation of cultures with calpain inhibitor I or calpeptin (100 microM). Furthermore, NMDA exposure decreased the number of GluR1 subunits of AMPA receptors detected by an antibody against the C-terminal domain of the subunit in western blots and resulted in the formation of a lower molecular weight species detected by an antibody against the N-terminal domain. Both effects were completely prevented by calpain inhibitors. These results indicate that NMDA receptor activation produces calpain activation and complex modifications of AMPA receptor properties, which could be involved in NMDA receptor-mediated changes in synaptic efficacy. PMID- 9202304 TI - GABA(A) receptor blockade in the anterior ventral tegmental area increases extracellular levels of dopamine in the nucleus accumbens of rats. AB - Previously, it was shown that microinfusion of the GABA(A) antagonist picrotoxin into the anterior ventral tegmental area (VTA) is reinforcing. It was hypothesized that this reinforcing effect of picrotoxin in the anterior VTA is mediated, at least in part, by the activation of the mesoaccumbens dopamine (DA) system. The objective of the present study was to determine if blockade of GABA(A) receptors in the anterior VTA can increase extracellular levels of DA in the nucleus accumbens (ACB), using an in vivo microdialysis technique in freely moving rats. Concentrations of picrotoxin (40, 80, and 160 microM) that had previously been shown to produce a reinforcing effect increased the extracellular levels of DA and its major metabolites in the ACB. The increased extracellular DA levels induced by intra-VTA injection of picrotoxin was markedly attenuated by coadministration with the GABA(A) agonist muscimol, whereas intra-VTA injection of muscimol alone did not have an apparent effect on extracellular DA levels in the ACB. Microinjection of another GABA(A) antagonist, bicuculline, into the anterior VTA also increased the extracellular release of DA in the ACB. These results suggest that DA neurons projecting from the anterior VTA to the ACB are tonically inhibited by GABA through its actions at the GABA(A) receptors. PMID- 9202305 TI - Impact of corticotropin-releasing hormone on extracellular norepinephrine in prefrontal cortex after chronic cold stress. AB - We have previously demonstrated that exposing rats to cold (5 degrees C) for 3-4 weeks potentiates the increase in extracellular norepinephrine (NE) in the medial prefrontal cortex produced by acute tail shock. In the present study, we used microdialysis to determine the duration of cold exposure required to produce this sensitization and explored the mechanism of the phenomenon. Tail shock elicited a twofold greater increase in extracellular NE in the medial prefrontal cortex of rats exposed to cold for 2 weeks than in naive control rats or in rats exposed to cold for 1 week and tested either immediately or after a 2-week delay. Local infusion of 10 microM D-amphetamine or 30 mM K+ increased extracellular NE in the medial prefrontal cortex (approximately 350 and 190%, respectively) comparably in control rats and rats exposed to cold for 3 weeks. In contrast, intraventricular administration of 3.0 microg of corticotropin-releasing hormone increased extracellular NE in the medial prefrontal cortex by 65% in rats exposed to cold for 2 weeks, but only 35% in control rats. These results indicate that an enhanced responsiveness of noradrenergic neurons to acute tail shock (1) requires approximately 2 weeks of cold exposure to develop and (2) may be mediated by a change at the level of the noradrenergic cell bodies rather than the nerve terminals. PMID- 9202306 TI - Metabotropic glutamate receptor mGluR5 in astrocytes: pharmacological properties and agonist regulation. AB - Metabotropic glutamate receptor (mGluR) agonists induce extensive phosphoinositide (PI) hydrolysis in astrocytes grown in a chemically defined medium with select growth factors. These astrocytes express mGluR5 transcripts, but none of the splice variants of mGluR1, thus permitting the characterization of mGluR5 in a native CNS cell without interference from mGluR1 activity. mGluR5 activation (1) was not associated with stimulation of cyclic AMP formation, (2) showed high sensitivity to the removal of extracellular versus intracellular Ca2+, (3) displayed high coupling efficiency relative to receptor density, and (4) induced PI hydrolysis that was suppressed by phorbol esters with low potency. The rank order of agonist potency was similar to that observed in mGluR1 and mGluR5 transfected cells. The phenylglycine antagonists tested were effective in blocking responses to 1-aminocyclopentane-1S,3R-dicarboxylic acid, but not to glutamate. Prolonged exposure to agonists induced a two-phase desensitization of mGluR5 function, an initial phase (completed by 1 h and plateaus for another 3 h) and a late phase (progressive decrease to approximately 30% of control levels by 24 h). Only the latter phase was associated with receptor down-regulation. Desensitization of mGluR5 function did not involve receptor internalization or phosphorylation mediated by protein kinase C or A; it was purely homologous, and reversible. Resensitization after short agonist treatment did not require prior receptor sequestration. Recovery after prolonged agonist exposure required new protein synthesis, but the restoration of function was more rapid than normalization of receptor protein levels, indicating that regulation also involves other components of the transduction system. The protracted desensitization of mGluR5 in astrocytes suggests that the functions mediated by this receptor are maintained under a variety of conditions ranging from repetitive stimulation to injury responses. PMID- 9202307 TI - Importance of the Rab3a-GTP binding domain for the intracellular stability and function of Rabphilin3a in secretion. AB - We had previously demonstrated that Rab3a-GTP inhibits and the Rab3a-binding protein Rabphilin3a enhances secretion in bovine chromaffin cells. In this study, we investigated the role of Rab3a-GTP binding in the intracellular expression and the function of Rabphilin3a in regulated exocytosis in bovine chromaffin cells. Using transient transfections, we found that a minimal domain, Rp(51-190), that inhibits secretion coincides with a minimal domain that effectively binds Rab3a GTP and allows intracellular stability of the construct. This domain includes a cysteine-rich, Zn2+-binding domain whose integrity is also required for Rab3a-GTP binding and the ability to inhibit secretion. A Rabphilin3a mutant, containing both C2 domains but defective in Rab3a-GTP, and wild-type Rabphilin3a both localized to chromaffin granules and stimulated secretion similarly despite lessened intracellular expression of the mutant protein. The data are consistent with a sequence of events in which a Rab3a-GTP x Rabphilin3a complex forms on the secretory granule as a precursor in a pathway that enhances secretion. The complex dissociates (perhaps because of GTP hydrolysis) to permit the enhancement of secretion by Rabphilin3a. PMID- 9202308 TI - N-acetylaspartylglutamate selectively activates mGluR3 receptors in transfected cells. AB - In previous studies, we demonstrated that the neuropeptide, N acetylaspartylglutamate (NAAG), meets the traditional criteria for a neurotransmitter and selectively activates metabotropic glutamate receptor mGluR2 or mGluR3 in cultured cerebellar granule cells and glia. Sequence homology and pharmacological data suggest that these two receptors are highly related structurally and functionally. To define more rigorously the receptor specificity of NAAG, cloned rat cDNAs for mGluR1-6 were transiently or stably transfected into Chinese hamster ovary cells and human embryonic kidney cells and assayed for their second messenger responses to the two endogenous neurotransmitters, glutamate and NAAG, as well as to metabotropic receptor agonists, trans-1 aminocyclopentane-1,3-dicarboxylate (trans-ACPD) and L-2-amino-4 phosphonobutyrate (L-AP4). Despite the high degree of relatedness of mGluR2 and mGluR3, NAAG selectively activated the mGluR3 receptor. NAAG activated neither mGluR2 nor mGluR1, mGluR4, mGluR5, or mGluR6. The mGluR agonist, trans-ACPD, activated each of the transfected receptors, whereas L-AP4 activated mGluR4 and mGluR6, consistent with the published selectivity of these agonists. Hybrid cDNA constructs of the extracellular domains of mGluR2 and mGluR3 were independently fused with the transmembrane and cytoplasmic domain of mGluR1a. This latter receptor domain is coupled to phosphoinositol turnover, and its activation increases intracellular calcium. The cells transfected with these chimeric receptors responded to activation by glutamate and trans-ACPD with increases in intracellular calcium. NAAG activated the chimeric receptor that contained the extracellular domain of mGluR3 and did not activate the mGluR2 chimera. PMID- 9202309 TI - BIMG 80, a novel potential antipsychotic drug: evidence for multireceptor actions and preferential release of dopamine in prefrontal cortex. AB - In radioligand binding studies, BIMG 80, a new putative antipsychotic, displayed good affinity at certain serotonin (5-HT1A, 5-HT2A, 5-HT6), dopamine (D1, D2L, D4), and noradrenergic (alpha1) receptors. The effect of acute subcutaneous BIMG 80, clozapine, haloperidol, risperidone, amperozide, olanzapine, and Seroquel was then investigated on dopamine release in medial prefrontal cortex, nucleus accumbens, and striatum in freely moving rats using the microdialysis technique. Four different neurochemical profiles resulted from the studies: (a) Systemic administration of BIMG 80, clozapine, and amperozide produced greater percent increases in dopamine efflux in medial prefrontal cortex than in the striatum or the nucleus accumbens. (b) Haloperidol induced a similar increase in dopamine concentrations in the striatum and nucleus accumbens with no effect in the medial prefrontal cortex. (c) Risperidone and olanzapine stimulated dopamine release to a similar extent in all brain regions investigated. (d) Seroquel failed to change significantly dopamine output both in the medial prefrontal cortex and in the striatum. Because an increase in dopamine release in the medial prefrontal cortex may be predictive of effectiveness in treating negative symptoms and in the striatum may be predictive of induction of extrapyramidal side effects, BIMG 80 appears to be a potential antipsychotic compound active on negative symptoms of schizophrenia with a low incidence of extrapyramidal side effects. PMID- 9202310 TI - Reactivating kinase/p38 phosphorylates tau protein in vitro. AB - Neurofibrillary tangles, one of the major pathological hallmarks of Alzheimer diseased brains, consist primarily of aggregated paired helical filaments (PHFs) of hyperphosphorylated tau protein. Tau from normal brain and especially from foetal brain is also phosphorylated on some of the sites phosphorylated in PHFs, mainly at serines or threonines followed by prolines. A number of protein kinases can phosphorylate tau in vitro; those that require or accept prolines include GSK3 and members of the mitogen-activated protein (MAP) kinase family, ERK1, ERK2, and SAP kinase-beta/JNK. In this report, we show that another member of the MAP kinase family, the stress-activated kinase p38/RK, can phosphorylate tau in vitro. Western blots with phosphorylation-sensitive antibodies showed that p38, like ERK2 and SAP kinase-beta/JNK, phosphorylated tau at sites found phosphorylated physiologically (Thr181, Ser202, Thr205, and Ser396) and also at Ser422, which is phosphorylated in neurofibrillary tangles but not in normal adult or foetal brain. These findings support the possibility that cellular stress might contribute to tau hyperphosphorylation during the formation of PHFs, and hence, to the development of tau pathology. PMID- 9202311 TI - Delayed phospholipid degradation in rat brain after traumatic brain injury. AB - Lipid second messengers such as arachidonic acid and its metabolites and diacylglycerols (DAGs) are affected in brain injury. Therefore, changes in the pool size and the fatty acid composition of free fatty acids (FFAs) and DAGs were analyzed in different rat brain areas 4 and 35 days after traumatic injury. Cortical impact injury of low-grade severity was applied in the right frontal somatosensory cortex. Four days after injury, FFAs and DAGs were increased by three- and twofold, respectively, in the injured cortex and to a lesser extent in the contralateral cortex compared with sham-operated animals. Docosahexaenoic acid followed by stearic acid, and arachidonic acid, displayed the greatest changes in both FFAs and DAGs. By day 35, free stearic, oleic, and arachidonic acids remained elevated in the damaged cortex (1.5-fold each). DAGs showed the greatest change, reaching values 2.7-fold higher than sham in all frontal and occipital cortical areas, including brainstem. Oleoyl- and arachidonoyl-DAGs (four- and threefold increase, respectively) followed by docosahexaenoyl-DAGs (twofold) contributed to the DAG accumulation. These results reveal that traumatic brain injury triggers a sustained and time-dependent activation of phospholipase-mediated signaling pathways leading to membrane phospholipid degradation and targeting, early on, docosahexaenoyl phospholipid-enriched excitable membranes. PMID- 9202312 TI - Acute and chronic treatments with citalopram lower somatostatin levels in rat brain striatum through different mechanisms. AB - The suggestion that somatostatin is involved in the pathophysiology of obsessive compulsive disorder and the evidence that selective serotonin reuptake inhibitors show significant antiobsessional effect prompted us to examine the effect of citalopram, a selective and potent serotonin reuptake inhibitor, on the somatostatinergic system in different brain regions of the rat. A single intraperitoneal injection of 10 mg/kg citalopram significantly reduced somatostatin levels in the striatum and nucleus accumbens after 4 but not 1, 8, or 24 h. No changes were found in hippocampus. In addition, we found that the K+ evoked overflow of somatostatin-like immunoreactivity from striatal slices was significantly increased 1 h after a single injection of citalopram and was still higher, although not significantly, 4 h after the drug injection. Levels of preprosomatostatin mRNA were unchanged in striatum and accumbens 1 and 4 h after a single drug administration. In rats treated with citalopram (10 mg/kg i.p.) twice daily for 14 days, the levels of somatostatin and its mRNA were significantly decreased in the striatum but not in other brain regions 24 h after the last dose. No change was found in the basal or K+-evoked overflow of somatostatin-like immunoreactivity at 1, 4, and 24 h after the last drug injection. These results suggest that acute and chronic treatment with citalopram reduces somatostatin levels in striatum by different mechanisms. Whereas a single dose of the drug reduces somatostatin levels by increasing the release of the peptide, repeated drug treatment reduces the biosynthesis of somatostatin. PMID- 9202313 TI - Demyelinating antibodies to myelin oligodendrocyte glycoprotein and galactocerebroside induce degradation of myelin basic protein in isolated human myelin. AB - Although the specificity of multiple sclerosis (MS) brain immunoglobulins (Igs) remains unknown, the incubation of these Igs with human myelin can lead to myelin basic protein (MBP) degradation mediated by neutral proteases. In this study, we demonstrate that monoclonal antibodies (mAbs) specific to myelin components such as the CNS-specific myelin oligodendrocyte glycoprotein (MOG) and galactocerebroside (GalC) are found to induce a significant loss of MBP mediated by neutral proteases in myelin. By contrast, antibodies to periaxonal and structural components of myelin, such as MBP and myelin-associated glycoprotein, are ineffective in inducing such MBP degradation. Among the 11 different anti-MOG mAbs directed to externally located epitopes of MOG, only two were found to induce a significant degradation of MBP, suggesting that antibody-induced MBP degradation is not only antigen specific but also epitope specific. Based on the inhibition of MBP degradation in the presence of EGTA and the analysis of the degradation products obtained following incubation of myelin with mAbs to GalC and MOG (8-18C5), the neutral protease involved in this antibody-induced degradation of MBP could be calcium-activated neutral protease. Taken together, these results suggest that antibodies to GalC and MOG can play a major role in destabilizing myelin through MBP breakdown mediated by neutral proteases and thus have an important role to play in the pathogenesis of MS. PMID- 9202314 TI - Expression of Bcl-2 in adult human brain regions with special reference to neurodegenerative disorders. AB - The expression of the protooncogene bcl-2, an inhibitor of apoptosis in various cells, was examined in the adult human brain. Several experimental criteria were used to verify its presence; mRNA was analyzed by northern blot with parallel experiments in mouse tissues, by RNase protection, and by in situ hybridization histochemistry. Bcl-2 protein was detected by western blot analysis and immunohistochemistry. Two bcl-2 mRNA species were identified in the human brain. The pattern of distribution of bcl-2 mRNA at the cellular level showed labeling in neurons but not glia. The in situ hybridization signal was stronger in the pyramidal neurons of the cerebral cortex and in the cholinergic neurons of the nucleus basalis of Meynert than in the Purkinje neurons of the cerebellum. Both melanized and nonmelanized neurons were labeled in the substantia nigra. In the striatum, bcl-2 mRNA was detected in some but not all neurons. In the regions examined for Bcl-2 protein, the expression pattern correlated with the mRNA results. In patients with Alzheimer's and Parkinson's diseases, quantification of bcl-2 mRNA in the nucleus basalis of Meynert and substantia nigra, respectively, showed that the expression was unaltered compared with controls, raising the possibility that the expression of other components of apoptosis is modulated. PMID- 9202315 TI - Early detection of DNA strand breaks in the brain after transient focal ischemia: implications for the role of DNA damage in apoptosis and neuronal cell death. AB - Using in situ DNA polymerase I-mediated biotin-dATP nick-translation (PANT) and terminal deoxynucleotidyl-transferase-mediated dUTP nick end-labeling (TUNEL), we investigated the evolution of DNA strand breaks, a marker of DNA damage, in rat brain after 1 h of middle cerebral artery occlusion and various durations of reperfusion. DNA single-strand breaks (SSBs) detected by PANT were present in neurons after as little as 1 min of reperfusion. Numbers of neurons containing an SSB increased progressively in the ischemic core but decreased in the ischemic penumbra after 1 h of reperfusion. DNA double-strand breaks (DSBs) detected by TUNEL were first seen in neurons after 1 h of reperfusion, and their numbers then increased progressively in the ischemic core, with a regional distribution similar to that of SSBs. However, the number of SSB-containing cells was greater than that of DSB-containing cells at all time points tested. SSB-containing cells detected within the first hour of reperfusion were exclusively neuronal and exhibited normal nuclear morphology. At 16-72 h of reperfusion, many SSB- and DSB containing cells, including both neurons and astrocytes, showed morphological changes consistent with apoptosis. Gel electrophoresis of DNA isolated from the ischemic core showed DNA fragmentation at 24 h, when both SSBs and DSBs were present, but not at 1 h, when few DSBs were detected. These results suggest that damage to nuclear DNA is an early event after neuronal ischemia and that the accumulation of unrepaired DNA SSBs may contribute to delayed ischemic neuronal death, perhaps by triggering apoptosis. PMID- 9202316 TI - Loss of the xeroderma pigmentosum group A gene (XPA) enhances apoptosis of cultured cerebellar neurons induced by UV but not by low-K+ medium. AB - To study the involvement of the xeroderma pigmentosum group A gene (XPA) in neuronal apoptosis, we cultured cerebellar neurons from mice lacking XPA gene (XPA-/-) and induced apoptosis by exposure to UV irradiation or medium containing a low concentration of potassium (low-K+ medium). When cerebellar neurons from postnatal days 15-16 wild-type mice were treated with UV irradiation, apoptotic neuronal death was observed after 24-48 h. About 60% of neurons survived 48 h after UV irradiation at a dose of 5 J/m2. On the other hand, neurons from XPA-/- mice showed a significantly increased vulnerability to UV irradiation, and >90% of neurons died 48 h after UV irradiation at a dose of 5 J/m2. In contrast, low K+ medium induced apoptosis of neurons from mice of each genotype with the same kinetics. These results suggest that the XPA gene is involved in neuronal DNA repair and that it thereby influences apoptosis induced by DNA damage in cultured cerebellar neurons. PMID- 9202317 TI - Amyloid beta protein (25-35) stimulation of phospholipase C in LA-N-2 cells. AB - The amyloid beta protein (25-35) stimulated appearance of 3H-inositol phosphates from [3H]inositol-prelabeled LA-N-2 cells was investigated. This stimulation was unaltered by extra- and intracellular calcium chelators in a calcium-free medium or by several protein kinase inhibitors. This phospholipase C stimulation by amyloid beta protein appeared to be pertussis toxin sensitive. It is possible that this phospholipase C stimulation by amyloid beta protein is a receptor mediated process. This possibility is based on two related observations. The stimulation is ablated by the presence of conventional antagonists for metabotropic, adrenergic, and bombesin agonists. The IC50 values were 12 microM for propranolol, 15 microM for AP-3, and 25 nM for [Tyr4,D-Phe12]bombesin. Additional support comes from results of desensitization and resensitization experiments. Amyloid beta protein stimulation of phospholipase C was absent from LA-N-2 cells previously treated with norepinephrine, trans-1-amino-1,3 cyclopentanedicarboxylic acid (t-ACPD), bombesin, or amyloid beta peptide. In a similar manner, LA-N-2 cells previously treated with amyloid beta protein were no longer responsive to norepinephrine, t-ACPD, or bombesin. The responsiveness to amyloid beta protein returned, subsequent to a period of resensitization for the individual agonists. It is suggested that this observed amyloid beta protein stimulation of phospholipase C may be responsible for the elevated quantity of inositol seen in the brains of Alzheimer's disease patients. PMID- 9202318 TI - S-adenosylmethionine decarboxylase activity is decreased in the rat cortex after traumatic brain injury. AB - S-Adenosyl-L-methionine decarboxylase (SAMdc) and L-ornithine decarboxylase (ODC) are major enzymes regulating polyamine synthesis. Following ischemia, putrescine content increases as a result of posttraumatic activation of ODC and inhibition of SAMdc. These alterations are thought to mediate edema and cell death. The purpose of this study was to quantify SAMdc activity and edema in the brain following controlled cortical impact injury. Anesthetized adult male rats underwent a right parietal craniectomy and were subjected to cortical impact injury. Tissues were obtained from three bilateral regions: parietal cortex, motor area (CPm); parietal cortex, somatosensory area (CPs); and the pyriform cortex (CPF). SAMdc activity was determined in the postmitochondrial fraction from homogenates of fresh, unfrozen tissues by measuring the decarboxylation of S adenosyl-L-[carboxyl-14C]methionine. Basal SAMdc activity was determined in unoperated rats, and regional differences were noted: Activity was lower in the CPF than in the CPm and CPs. SAMdc activity decreased to the greatest extent in the ipsilateral CPm (impact site) from 1 to 72 h following traumatic brain injury. Significant edema was found in the ipsilateral CPm 1, 8, 16, 24, and 48 h after injury. Decreased SAMdc activity impairs the conversion of putrescine to polyamines and may contribute to delayed pathological changes in the brain after traumatic injury. PMID- 9202319 TI - Aggregation of beta-amyloid peptide is promoted by membrane phospholipid metabolites elevated in Alzheimer's disease brain. AB - Increased amounts of beta-amyloid (A beta) peptide deposits are found in Alzheimer's disease brain. These amyloid deposits have been implicated in the pathophysiology of this common dementing illness. A beta peptides have been shown to be toxic to neurons in cell culture, and this toxicity is critically dependent on the aggregation of the peptide into cross-beta-pleated sheet fibrils. Also, in vivo and postmortem NMR studies have shown changes in certain brain membrane phospholipid metabolites in normal aging and more extensive alterations in patients with Alzheimer's disease. The finding that membrane phospholipids affect the aggregation of A beta suggests that the abnormalities in membrane metabolism found in Alzheimer's disease could affect the deposition of A beta in vivo. Therefore, we examined the effect of membrane phospholipid metabolites that are altered in Alzheimer's disease brain on the aggregation of A beta(1-40) using a light scattering method. Certain metabolites (glycerophosphocholine, glycerophosphoethanolamine, and alpha-glycerophosphate) augment the aggregation of A beta. Other membrane phospholipid metabolites (phosphocholine, phosphoethanolamine, and inositol-1-phosphate) have no effect. We conclude that increased membrane phospholipid metabolite concentrations may play a role in the deposition of A beta seen in normal aging and the even greater deposition of A beta observed in Alzheimer's disease. PMID- 9202320 TI - Impairment of glucose and glutamate transport and induction of mitochondrial oxidative stress and dysfunction in synaptosomes by amyloid beta-peptide: role of the lipid peroxidation product 4-hydroxynonenal. AB - Deposits of amyloid beta-peptide (A beta), reduced glucose uptake into brain cells, oxidative damage to cellular proteins and lipids, and excitotoxic mechanisms have all been suggested to play roles in the neurodegenerative process in Alzheimer's disease. Synapse loss is closely correlated with cognitive impairments in Alzheimer's disease, suggesting that the synapse may be the site at which degenerative mechanisms are initiated and propagated. We report that A beta causes oxyradical-mediated impairment of glucose transport, glutamate transport, and mitochondrial function in rat neocortical synaptosomes. A beta induced membrane lipid peroxidation in synaptosomes that occurred within 1 h of exposure; significant decreases in glucose transport occurred within 1 h of exposure to A beta and decreased further with time. The lipid peroxidation product 4-hydroxynonenal conjugated to synaptosomal proteins and impaired glucose transport; several antioxidants prevented A beta-induced impairment of glucose transport, indicating that lipid peroxidation was causally linked to this adverse action of A beta. FeSO4 (an initiator of lipid peroxidation), A beta, and 4 hydroxynonenal each induced accumulation of mitochondrial reactive oxygen species, caused concentration-dependent decreases in 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetrazolium bromide reduction, and reduced cellular ATP levels significantly. A beta also impaired glutamate transport, an effect blocked by antioxidants. These data suggest that A beta induces membrane lipid peroxidation, which results in impairment of the function of membrane glucose and glutamate transporters, altered mitochondrial function, and a deficit in ATP levels; 4 hydroxynonenal appears to be a mediator of these actions of A beta. These data suggest that oxidative stress occurring at synapses may contribute to the reduced glucose uptake and synaptic degeneration that occurs in Alzheimer's disease patients. They further suggest a sequence of events whereby oxidative stress promotes excitotoxic synaptic degeneration and neuronal cell death in a variety of different neurodegenerative disorders. PMID- 9202321 TI - Iodoacetate produces striatal excitotoxic lesions. AB - Impairment of energy production may play a role in the pathogenesis of Huntington's disease (HD). It was recently shown that huntingtin can bind to and possibly inhibit the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). We found that intrastriatal administration of the GAPDH inhibitor iodoacetate produces striatal lesions that are significantly attenuated by removal of the corticostriatal glutamatergic input, consistent with an excitotoxic mechanism. The lesions are accompanied by increased production of hydroxyl free radicals as assessed by conversion of salicylate to 2,3- and 2,5 dihydroxybenzoic acid. In vivo magnetic resonance imaging showed lesions on T2 weighted scans, but there was only a small increase in lactate content. These results show that inhibition of GAPDH produces striatal lesions in vivo and suggest that inhibition of GAPDH could contribute to neuronal degeneration in HD. PMID- 9202322 TI - Brain-specific modulation of kynurenic acid synthesis in the rat. AB - This study was designed to investigate modulatory mechanisms that control the synthesis of the neuroprotective endogenous excitatory amino acid receptor antagonist kynurenate. De novo kynurenate formation was examined in vitro using tissue slices from rat brain, liver, and kidney. In slices from adult cerebral cortex, veratridine, quisqualate, and L-alpha-aminoadipate decreased kynurenate synthesis substantially. Glucose removal or changes in the ionic milieu, too, influenced kynurenate formation significantly, suggesting that demands on cellular energy interfere with kynurenate production in the adult rat brain. The effects of quisqualate and L-alpha-aminoadipate were also observed in the immature brain, in the quinolinate-lesioned adult striatum, and, to a lesser extent, in peripheral organs. In contrast, the effect of veratridine was not seen in the lesioned brain or in kidney and liver tissue, indicating its dependency on intact neuron-glia interactions. Compared with the normal adult brain, ionic manipulations yielded qualitatively distinct results in the developing brain and in the periphery, but their effects remained unchanged in the lesioned striatum. Glucose deprivation was less consequential in the immature than in the adult brain and was entirely ineffective in the lesioned striatum and in the periphery. These results further link cellular, especially astrocytic, energy metabolism to kynurenate formation in the brain. More generally, the existence of brain specific mechanisms for the regulation of kynurenate production is suggestive of a modulatory role of this metabolite in excitatory amino acid receptor function and dysfunction. PMID- 9202323 TI - alpha2-Macroglobulin as a beta-amyloid peptide-binding plasma protein. AB - The beta-amyloid peptide (A beta) is a normal proteolytic processing product of the amyloid precursor protein, which is constitutively expressed by many, if not most, cells. For reasons that are still unclear, A beta is deposited in an aggregated fibrillar form in both diffuse and senile plaques in the brains of patients with Alzheimer's disease (AD). The factor(s) responsible for the clearance of soluble A beta from biological fluids or tissues are poorly understood. We now report that human alpha2-macroglobulin (alpha2M), a major circulating endoproteinase inhibitor, which has recently been shown to be present in senile plaques in AD, binds 125I-A beta(1-42) with high affinity (apparent dissociation constant of 3.8 x 10(-10) M). Approximately 1 mol of A beta is bound per mole of alpha2M. Both native and methylamine-activated alpha2M bind 125I-A beta(1-42). The binding of 125I-A beta(1-42) to alpha2M is enhanced by micromolar concentrations of Zn2+ (but not Ca2+) and is inhibited by noniodinated A beta(1 42) and A beta(1-40) but not by the reverse peptide A beta(40-1) or the cytokines interleukin 1beta or interleukin 2. alpha1-Antichymotrypsin, another plaque associated protein, inhibits both the binding of 125I-A beta(1-42) to alpha2M as well as the degradation of 125I-A beta(1-42) by proteinase-activated alpha2M. Moreover, the binding of 125I-A beta(1-42) to alpha2M protects the peptide from proteolysis by exogenous trypsin. These data suggest that alpha2M may function as a carrier protein for A beta and could serve to either facilitate or impede clearance of A beta from tissues such as the brain. PMID- 9202324 TI - AP-1 and Egr DNA-binding activities are increased in rat brain during ethanol withdrawal. AB - The DNA-binding activities of AP-1 and Egr proteins were investigated in nuclear extracts of rat brain regions during ethanol withdrawal. Both DNA-binding activities were transiently elevated in the hippocampus and cerebellum 16 h after withdrawal. In the cerebral cortex, AP-1 and Egr DNA-binding activities increased at 16 h and persisted until 32 and 72 h, respectively. The AP-1 DNA-binding activities in all regions at all times after withdrawal were composed of FosB, c Jun, JunB, and JunD. c-Fos was detected at all times in the cerebral cortex, at 16 h only in the hippocampus, and from 16 to 72 h in the cerebellum. Withdrawal severity did not affect the composition of the AP-1 DNA-binding activities. Two Egr DNA-binding activities were present in the cortex and hippocampus. The faster migrating complex predominated in hippocampus, and only the slower-migrating complex (identified as Egr-1) was present in the cerebellum. The increase in DNA binding activity of immediate early gene-encoded transcription factors supports their proposed role in initiating a cascade of altered gene expression underlying the long-term neuronal response to ethanol withdrawal. PMID- 9202325 TI - Desensitisation of the adenosine A1 receptor by the A2A receptor in the rat striatum. AB - The influence of the adenosine A2A receptor on the A1 receptor was examined in rat striatal nerve terminals, a model for other cells in which these receptors are coexpressed. Incubation of striatal synaptosomes with the A2A receptor agonist 2-p-(2-carboxyethyl)phenethylamino-5'-N-ethylcarboxamidoadenosine (CGS 21680) caused the appearance of a low-affinity binding site for the A1 receptor agonist 2-chloro-N6-cyclopentyladenosine (CCPA). This effect was blocked by the A2A receptor antagonist ZM241385 and by the protein kinase C inhibitor chelerythrine, but not by the protein kinase A inhibitor N-(2-guanidinoethyl)-5 isoquinolinesulfonamide (HA 1004). The effect was not seen with striatal membranes or with hypotonically lysed synaptosomes. These results demonstrate a protein kinase C-mediated heterologous desensitisation of the A1 receptor by the A2A receptor. PMID- 9202326 TI - An endogenous dopaminergic neurotoxin, N-methyl-(R)-salsolinol, induces DNA damage in human dopaminergic neuroblastoma SH-SY5Y cells. AB - Recently, an endogenous neurotoxin, 1(R),2(N)-dimethyl-6,7-dihydroxy-1,2,3,4 tetrahydroisoquinoline [N-methyl-(R)-salsolinol], was found to elicit parkinsonism in rats with selective depletion of dopamine neurons in the substantia nigra without necrotic tissue reaction. The mechanism of the cell death was examined by detection of DNA damage using a single-cell gel electrophoresis (comet) assay in human dopaminergic neuroblastoma SH-SY5Y cells. Only N-methylsalsolinol was found to induce DNA damage, whereas other catechol isoquinolines, such as (R)-salsolinol, (S)-salsolinol, and 1,2-dimethyl-6,7 dihydroxyisoquinolinium ion, did not. The (R)-enantiomer of N-methylsalsolinol damaged DNA much more profoundly than the (S)-enantiomer. Cycloheximide protected the cells from DNA damage, suggesting that an apoptotic process may account for the DNA damage. Morphological changes indicating apoptotic cell death were also confirmed. Antioxidants and deprenyl reduced DNA damage, indicating that the damage was initiated by oxidative stress and that neuroprotection by deprenyl may be partially ascribed to its prevention of DNA damage. Apoptosis induced by neurotoxins may be a mechanism underlying the cell death of dopamine neurons in the substantia nigra of Parkinson's disease. PMID- 9202327 TI - Axonal contact regulates expression of alpha2 and beta2 isoforms of Na+, K+ ATPase in Schwann cells: adhesion molecules and nerve regeneration. AB - Three isoforms of catalytic alpha subunits and two isoforms of beta subunits of Na+,K+-ATPase were detected in rat sciatic nerves by western blotting. Unlike the enzyme in brain, sciatic nerve Na+,K+-ATPase was highly resistant to ouabain. The ouabain-resistant alpha1 isoform was demonstrated to be the predominant form in rat intact sciatic nerve by quantitative densitometric analysis and is mainly responsible for sciatic nerve Na+,K+-ATPase activity. After sciatic nerve injury, the alpha3 and beta1 isoforms completely disappeared from the distal segment owing to Wallerian degeneration. In contrast, alpha2 and beta2 isoform expression and Na+,K+-ATPase activity sensitive to pyrithiamine (a specific inhibitor of the alpha2 isoform) were markedly increased in Schwann cells in the distal segment of the injured sciatic nerve. These latter levels returned to baseline with nerve regeneration. Our results suggest that alpha3 and beta1 isoforms are exclusive for the axon and alpha2 and beta2 isoforms are exclusive for the Schwann cell, although axonal contact regulates alpha2 and beta2 isoform expressions. Because the beta2 isoform of Na+,K+-ATPase is known as an adhesion molecule on glia (AMOG), increased expression of AMOG/beta2 on Schwann cells in the segment distal to sciatic nerve injury suggests that AMOG/beta2 may act as an adhesion molecule in peripheral nerve regeneration. PMID- 9202329 TI - A novel cdc2-related protein kinase expressed in the nervous system. AB - We report the cloning and characterization of a cDNA encoding a cdc2-related protein kinase, named PFTAIRE, that is expressed primarily in the postnatal and adult nervous system. We have demonstrated by in situ hybridization and indirect immunofluorescence that several populations of terminally differentiated neurons and some neuroglia expressed PFTAIRE mRNA and protein. In neurons, PFTAIRE protein was localized in the nucleus and cytoplasm of cell bodies. The anatomical, cellular, and ontogenic patterns of PFTAIRE expression in the nervous system differed from those of p34cdc2 and cdk5, which are expressed in brain and several other mitotic tissues. Proteins of approximately 58-60 kDa coprecipitated specifically with PFTAIRE from cytosolic protein preparations of adult mouse brain and transfected cells. These proteins appeared to be the major endogenous substrates associated with this kinase activity. The temporal and spatial expression patterns of PFTAIRE in the postnatal and adult nervous system suggest that PFTAIRE kinase activity may be associated with the postmitotic and differentiated state of cells in the nervous system and that its function may be distinct from those of p34cdc2 and cdk5. PMID- 9202328 TI - L-aspartate but not the D form is secreted through microvesicle-mediated exocytosis and is sequestered through Na+-dependent transporter in rat pinealocytes. AB - Rat pinealocytes accumulate glutamate in microvesicles and secrete it through exocytosis so as to transmit signals intercellularly. Glutamate is involved in the negative regulation of norepinephrine-stimulated melatonin production. In this study, we found that aspartate is also released from cultured rat pinealocytes during the exocytosis of glutamate. The release of aspartate was triggered by addition of KCI or A23187 (a Ca2+ ionophore) in the presence of Ca2+ and was proportional to the amount of L-glutamate released. Furthermore, the release of aspartate was inhibited by both botulinum neurotoxin type E and L- or N-type voltage-gated Ca2+ channel blockers. Bay K 8644, an agonist for the L-type Ca2+ channel, stimulated the release of aspartate 2.1-fold. Immunohistochemical analyses with antibodies against aspartate and synaptophysin revealed that aspartate is colocalized with synaptophysin in a cultured pinealocyte. HPLC with fluorometric detection indicated that the released aspartate is of the L form, although pinealocytes also contain the D form in their cytoplasm, corresponding to approximately 30% of the total free aspartate. Radiolabeled L-aspartate was taken up by the microsomal fraction from bovine pineal glands in a Na+-dependent manner. The Na+-dependent uptake of L-aspartate was strongly inhibited by L cysteine sulfinate, beta-hydroxyaspartate, and L-serine-O-sulfate, inhibitors for the Na+-dependent glutamate/aspartate transporter on the plasma membrane. Na+ dependent sequestration of L-aspartate was also observed in cultured rat pinealocytes, which was inhibited similarly by these transporter inhibitors. These results strongly suggest that L-aspartate is released through microvesicle mediated exocytosis from pinealocytes and is taken up again through the Na+ dependent transporter at the plasma membrane. The possible role of L-aspartate as an intercellular chemical transmitter in the pineal gland is discussed. PMID- 9202331 TI - Distribution of ankyrin isoforms and their proteolysis after ischemia and reperfusion in rat brain. AB - The distribution of brain-type ankyrin (ankyrinB, 212 kDa) and erythrocyte-type ankyrin (ankyrinR, 239 kDa) was investigated in the subcellular fractions of rat forebrain (P1, 1,000 g pellet; P2, 15,000 g pellet; P3, 100,000 g pellet; S, 100,000 g supernatant) by immunoblotting using specific antibodies. The P2 fraction contained approximately 40% of the 212- and 163-kDa isoforms of ankyrinB and the 239-kDa isoform of ankyrinR. Further subfractionation of the P2 by Percoll gradient centrifugation followed by separation of myelin showed association of the three ankyrin isoforms with the synaptosome-rich fraction but not with the myelin-rich fraction. The plasma membrane-rich P3 fraction contained a concentration of ankyrin isoforms similar to that in the P2 fraction. In vitro proteolysis of ankyrin in the P2 fraction with calpain showed that the 212-kDa ankyrinB was more susceptible to calpain than was ankyrinR. In the two-vessel occlusion model, ischemia for 30 min generated the 160-kDa fragment of ankyrinR, and reperfusion for 60 min after 30 min of ischemia remarkably increased the 160 kDa fragment. The reperfusion also significantly decreased the 212-kDa isoform of ankyrinB. Both ischemia-reperfusion and in vitro proteolysis with calpain generated the 160-kDa fragment of ankyrinR, suggesting the involvement of calpain. PMID- 9202330 TI - In vivo evidence that GABA(B) receptors are negatively coupled to adenylate cyclase in rat striatum. AB - The characteristics of the cerebral GABA(B) receptor/cyclic AMP (cAMP)-generating system were investigated using the in vivo microdialysis technique in freely moving rats. Addition of forskolin, an activator of adenylate cyclase, to perfusate for 20 min resulted in a dose-dependent increase of cAMP efflux from the striatum. Pre- and coinfusions of baclofen for 80 min had no effect on the basal efflux of cAMP from the striatum but induced a significant decrease of forskolin (10 microM)-stimulated cAMP efflux from the striatum in a dose dependent manner. SKF 97541 (100 microM), a GABA(B) receptor agonist, and GABA (50 microM) also decreased forskolin-induced cAMP efflux from the striatum. Coinfusion of CGP 54626A (100 microM), a GABA(B) receptor antagonist, counteracted the effect of baclofen on the forskolin-stimulated cAMP efflux. In contrast, the isoproterenol (5 mM)-induced increase of cAMP efflux from the striatum was significantly enhanced by pre- and coinfusions with baclofen. These results suggest that this test system using in vivo microdialysis may be useful for examining the effect of drugs on the GABA(B) receptor-linked cAMP-generating system in vivo. PMID- 9202332 TI - Hypoxia-induced catecholamine release and intracellular Ca2+ increase via suppression of K+ channels in cultured rat adrenal chromaffin cells. AB - Hypoxia (5% O2) enhanced catecholamine release in cultured rat adrenal chromaffin cells. Also, the intracellular free Ca2+ concentration ([Ca2+]i) increased within 3 min in approximately 50% of the chromaffin cells under hypoxic stimulation. The increase depended on the presence of extracellular Ca2+. Nifedipine and omega conotoxin decreased the population of the cells that showed the hypoxia-induced [Ca2+]i increase, showing that the Ca2+ influx was attributable to L- and N-type voltage-dependent Ca2+ channels. The membrane potential was depolarized during the perfusion with the hypoxic solution and returned to the basal level following the change to the normoxic solution (20% O2). Membrane resistance increased twofold under the hypoxic condition. The current-voltage relationship showed a hypoxia-induced decrease in the outward K+ current. Among the K+ channel openers tested, cromakalim and levcromakalim, both of which interact with ATP-sensitive K+ channels, inhibited the hypoxia-induced [Ca2+]i increase and catecholamine release. The inhibitory effects of cromakalim and levcromakalim were reversed by glibenclamide and tolbutamide, potent blockers of ATP-sensitive K+ channels. These results suggest that some fractions of adrenal chromaffin cells are reactive to hypoxia and that K+ channels sensitive to cromakalim and glibenclamide might have a crucial role in hypoxia-induced responses. Adrenal chromaffin cells could thus be a useful model for the study of oxygen-sensing mechanisms. PMID- 9202333 TI - Aggregation state-dependent activation of the classical complement pathway by the amyloid beta peptide. AB - Activation of the classical complement pathway has been widely investigated in recent years as a potential mechanism for the neuronal loss and neuritic dystrophy characteristic of Alzheimer's disease (AD) pathogenesis. We have previously shown that amyloid beta peptide (A beta) is a potent activator of complement, and recent evidence suggesting that the assembly state of A beta is crucial to the progress of the disease prompted efforts to determine whether the ability of A beta to activate the classical complement pathway is a function of the aggregation state of the peptide. In this report, we show that the fibrillar aggregation state of A beta, as determined by thioflavin T fluorometry, electron microscopy, and staining with Congo red and thioflavine S, is precisely correlated with the ability of the peptide to induce the formation of activated fragments of the complement proteins C4 and C3. These results suggest that the classical complement pathway provides a mechanism whereby complement-dependent processes may contribute to neuronal injury in the proximity of fibrillar but not diffuse A beta deposits in the AD brain. PMID- 9202335 TI - Interactions between neuronal histamine and halothane anesthesia in rats. AB - Using an in vivo microdialysis method, we measured the release of histamine in the anterior hypothalamic area (AHy) of rats under several concentrations of halothane anesthesia (1, 0.5, and 0.2%). The release of histamine increased to 341 and 325% at halothane concentrations of 0.5 and 0.2%, compared with the basal level at anesthesia induced by 1% halothane. alpha-Fluoromethylhistidine (100 mg/kg i.v.), a specific and irreversible inhibitor of histidine decarboxylase, reduced the histamine release to <35% of the basal value at 1% halothane anesthesia in the AHy, and also decreased the anesthetic requirement for halothane, evaluated as the minimum alveolar concentration (MAC), by 26%. Furthermore, pyrilamine (20 mg/kg i.v.), a brain-penetrating H1 antagonist, and zolantidine (20 mg/kg i.v.), a brain-penetrating H2 antagonist, reduced the MAC for halothane by 28.5 and 16%, respectively. Although thioperamide (5 mg/kg i.v.), an antagonist of presynaptic H3 autoreceptor, induced an approximate twofold increase in the level of histamine release in conscious freely moving rats, the same dose of thioperamide had little effect on the release of histamine under 1% halothane anesthesia in the AHy. Furthermore, thioperamide did not change the anesthetic requirement (MAC) for halothane. The present findings indicate that halothane anesthesia inhibits the release of neuronal histamine and that histaminergic neuron activities change the anesthetic requirement (MAC) for halothane through H1 as well as H2 receptors. PMID- 9202336 TI - Lesioning of deep prepiriform cortex protects against ischemic neuronal necrosis by attenuating extracellular glutamate concentrations. AB - An area of the deep prepiriform cortex is a controlling site for limbic seizures. Focal pharmacologic blockade of NMDA receptors in the deep prepiriform cortex protects against hippocampal cell injury during limbic seizures induced by intravenous kainate and during the excitotoxicity of global ischemia. In the current study, the deep prepiriform cortex was lesioned bilaterally by microinjection of kainate, 3 days before 10 min of global ischemia induced by four-vessel occlusion. Extracellular glutamate concentrations in the hippocampus were measured before, during, and after global ischemia by using in vivo microdialysis technique. Surviving hippocampal neurons were counted 7 days after ischemia. Lesioned animals showed significantly greater numbers of surviving neurons and significantly lower ischemia-induced elevations of extracellular glutamate concentrations than nonlesioned animals. During seizures induced from the deep prepiriform cortex, the immediate early gene cox-2 is expressed in the hippocampus. These results indicate that deep prepiriform cortex can be a modulatory site for ischemic hippocampal injury. PMID- 9202334 TI - Characterization of 5,7-dichlorokynurenate-insensitive D-[3H]serine binding to synaptosomal fraction isolated from rat brain tissues. AB - To explore target sites for endogenous D-serine that are different from the glycine site of the N-methyl-D-aspartate (NMDA) type glutamate receptor, we have studied the binding of D-[3H]serine to the synaptosomal P2 fraction prepared from the rat brain and peripheral tissues in the presence of an excess concentration (100 microM) of the glycine site antagonist 5,7-dichlorokynurenate (DCK). Nonspecific binding was defined in the presence of 1 mM unlabeled D-serine. Association, dissociation, and saturation experiments indicated that D-[3H]serine bound rapidly and reversibly to a single population of recognition sites in the cerebellar P2 fraction in the presence of DCK, with a K(D) of 614 nM and a Bmax of 2.07 pmol/mg of protein. D-Serine, L-serine, and glycine produced a total inhibition of the specific DCK-insensitive D-[3H]serine binding to the cerebellum with similar Ki values. Strychnine and 7-chlorokynurenate failed to inhibit the binding at 10 microM. The profiles of displacement of the DCK-insensitive D [3H]serine binding by various amino acids and glutamate and glycine receptor related compounds differ from those of any other defined recognition sites. DCK insensitive D-[3H]serine binding was at high levels in the cerebral cortex and cerebellum but very low in the kidney and liver. The present findings indicate that the DCK-insensitive D-[3H]serine binding site could be a novel candidate for a target for endogenous D-serine in mammalian brains. PMID- 9202338 TI - Brain lactate is an obligatory aerobic energy substrate for functional recovery after hypoxia: further in vitro validation. AB - This study used the rat hippocampal slice preparation and the monocarboxylate transporter inhibitor, alpha-cyano-4-hydroxycinnamate (4-CIN), to assess the obligatory role that lactate plays in fueling the recovery of synaptic function after hypoxia upon reoxygenation. At a concentration of 500 microM, 4-CIN blocked lactate-supported synaptic function in hippocampal slices under normoxic conditions in 15 min. The inhibitor had no effect on glucose-supported synaptic function. Of control hippocampal slices exposed to 10-min hypoxia, 77.8 +/- 6.8% recovered synaptic function after 30-min reoxygenation. Of slices supplemented with 500 microM 4-CIN, only 15 +/- 10.9% recovered synaptic function despite the large amount of lactate formed during the hypoxic period and the abundance of glucose present before, during, and after hypoxia. These results indicate that 4 CIN, when present during hypoxia and reoxygenation, blocks lactate transport from astrocytes, where the bulk of anaerobic lactate is formed, to neurons, where lactate is being utilized aerobically to support recovery of function after hypoxia. These results unequivocally validate that brain lactate is an obligatory aerobic energy substrate for posthypoxia recovery of function. PMID- 9202337 TI - Essential active-site lysine of brain glutamate dehydrogenase isoproteins. AB - Two soluble forms of bovine brain glutamate dehydrogenase (GDH) isoproteins were inactivated by pyridoxal 5'-phosphate. Spectral evidence is presented to indicate that the inactivation proceeds through Schiff's base formation with amino groups of the enzyme. Sodium borohydride reduction of the pyridoxal 5'-phosphate inactivated GDH isoproteins produced a stable pyridoxyl enzyme derivative that could not be reactivated by dialysis. The pyridoxyl enzyme was studied through fluorescence spectroscopy. No substrates or coenzymes separately gave complete protection against pyridoxal 5'-phosphate. A combination of 10 mM 2-oxoglutarate with 2 mM NADH, however, gave complete protection against the inactivation. Tryptic peptides of the isoproteins, modified with and without protection, resulted in a selective modification of one lysine. In both GDH isoproteins, the sequences of the peptide containing the phosphopyridoxyllysine were clearly identical to sequences of other GDH species. PMID- 9202339 TI - Serotonin inhibition of the NMDA receptor/nitric oxide/cyclic GMP pathway in rat cerebellum: involvement of 5-hydroxytryptamine2C receptors. AB - Previous studies have shown that 5-hydroxytryptamine (5-HT) can potently inhibit glutamatergic transmission in rat cerebellum through the activation of multiple 5 HT receptors. The aim of this study was to subclassify the 5-HT2 receptor mediating inhibition of the cyclic GMP response elicited by N-methyl-D-aspartate in adult rat cerebellar slices. Seven receptor antagonists, endowed with relative selectivities for the 5-HT2A, 5-HT2B, and 5-HT2C subtypes, differentially affected the inhibition by (+/-)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane of the cyclic GMP response, suggesting that the receptor involved belongs to the 5 HT2C subtype. PMID- 9202340 TI - Polyglutamine domains are substrates of tissue transglutaminase: does transglutaminase play a role in expanded CAG/poly-Q neurodegenerative diseases? AB - Huntington's disease and six other neurodegenerative diseases are associated with abnormal gene products containing expanded polyglutamine (poly-Q; Qn) domains (n > or = 40). In the present work, we show that glutathione S-transferase (GST) fusion proteins containing a small, physiological-length poly-Q domain (GSTQ10) or a large, pathological-length poly-Q domain (GSTQ62) are excellent substrates of guinea pig liver (tissue) transglutaminase and that both GSTQ10 and GSTQ62 are activators of tissue transglutaminase-catalyzed hydroxaminolysis of N-alpha carbobenzoxyglutaminylglycine. The present findings have implications for understanding the pathophysiological mechanisms of expanded CAG/poly-Q domain diseases. PMID- 9202341 TI - Subcellular translocation of Ca2+/calmodulin-dependent protein kinase II: fact or artifact? PMID- 9202342 TI - The forgotten 5-hydroxyindoleacetic acid. PMID- 9202343 TI - Results of coronary artery bypass grafting by a single surgeon in patients with left ventricular ejection fractions < or = 30%. AB - Despite the ominous prognosis of severe left ventricular (LV) dysfunction from coronary artery disease, coronary artery bypass grafting (CABG) in this setting remains controversial because of concerns over high operative risk and low likelihood of functional or survival benefit. We analyzed 135 consecutive patients (113 men, 22 women; age 42 to 87 years, mean 66.5) with LV ejection fraction (EF) < or =30% undergoing isolated CABG by 1 surgeon over an 8-year period. LVEF ranged from 10% to 30% (mean 23.6%). Preoperatively, 63% of patients had angina, 61% had heart failure (23% with pulmonary edema), and 24% manifested severe ventricular arrhythmia. The mean number of grafts was 2.7 per patient. The internal mammary artery was used in 103 of the 120 grafts (86%) to the left anterior descending coronary artery. Seven patients (5.2%) died in hospital. Only 2 of 99 patients (2%) not in intensive care preoperatively died in hospital. Angina class improved by 2.0 categories and congestive heart failure class by 1.5 categories. LVEF (assessed in 104 of 128 hospital survivors) improved from 24% preoperatively to 34% postoperatively (p <0.0001). At 1, 3, and 4.5 years respectively, all-cause survival was 87%, 81%, and 71%, and freedom from cardiac death was 90%, 85%, and 80%. CABG in patients with coronary artery disease and advanced LV dysfunction: (1) can be performed relatively safely, (2) achieves good long-term survival, (3) improves LVEF, (4) improves quality of life, and (5) can safely utilize the internal mammary artery as a conduit. The use of CABG is encouraged for patients with advanced LV dysfunction and may provide a viable alternative to transplantation in selected patients. PMID- 9202344 TI - Value of posterior and right ventricular leads in comparison to the standard 12 lead electrocardiogram in evaluation of ST-segment elevation in suspected acute myocardial infarction. AB - In this multicenter prospective trial, we studied posterior (V7 to V9) and right ventricular (V4R to V6R) leads to assess their accuracy compared with standard 12 lead electrocardiograms (ECGs) for the diagnosis of acute myocardial infarction (AMI). Patients aged >34 years with suspected AMI received posterior and right ventricular leads immediately after the initial 12-lead ECG. ST elevation of 0.1 mV in 2 leads was blindly determined and inter-rater reliability estimated. AMI was diagnosed by World Health Organization criteria. The diagnostic value of nonstandard leads was determined when 12-lead ST elevation was absent and present and multivariate stepwise regression analysis was also performed. Of 533 study patients, 64.7% (345 of 533) had AMI and 24.8% received thrombolytic therapy. Posterior and right ventricular leads increased sensitivity for AMI by 8.4% (p = 0.03) but decreased specificity by 7.0% (p = 0.06). The likelihood ratios of a positive test for 12, 12 + posterior, and 12 + right ventricular ECGs were 6.4, 5.6, and 4.5, respectively. Increased AMI rates (positive predictive values) were found when ST elevation was present on 6 nonstandard leads (69.1%), on 12 leads only (88.4%), and on both 6 and 12 leads (96.8%; p <0.001). Treatment rates with thrombolytic therapy increased in parallel with this electrocardiographic gradient. Logistic regression analysis showed that 4 leads were independently predictive of AMI (p <0.001): leads I, II, V3, V5R; V9 approached statistical significance (p = 0.055). The standard ECG is not optimal for detecting ST segment elevation in AMI, but its accuracy is only modestly improved by the addition of posterior and right ventricular leads. PMID- 9202345 TI - Timing and mechanism of death determined clinically after primary angioplasty for acute myocardial infarction. AB - We reviewed the timing and mechanism of death in 1,184 consecutive patients with acute myocardial infarction (AMI) treated with primary angioplasty from 1984 to 1995. Of 98 deaths, 48 (49%) occurred early on day 0 or 1. The mechanisms of death were pump failure in 60 patients (61%), reinfarction in 7 patients (7.1%), left ventricular rupture in 5 patients (5.1%), arrhythmia in 3 patients (3.1%), other cardiac causes in 5 patients (5.1%), stroke in 6 patients (6.1%), anoxic encephalopathy in 7 patients (7.1%), and procedure-related deaths in 5 patients (5.1%). The strongest predictors of mortality were cardiogenic shock and unsuccessful reperfusion. Our data indicate that mortality after primary angioplasty, like thrombolytic therapy, is highest in the early hours and is usually due to pump failure. In contrast to thrombolytic therapy, the incidence of death from myocardial rupture and bleeding complications is low. Future treatment strategies will need to focus on the large number of patients with early death due to pump failure, especially patients with cardiogenic shock. PMID- 9202346 TI - One-week and six-month angiographic controls of stent implantation after occlusive and nonocclusive dissection during primary balloon angioplasty for acute myocardial infarction. AB - We prospectively assessed in 124 consecutive patients by means of 1-week and 6 month follow-up angiograms the rate of reocclusion and restenosis of coronary stenting with Palmaz-Schatz stents after occlusive and nonocclusive dissection during primary balloon angioplasty for acute myocardial infarction (AMI). Patients were further evaluated clinically at 1 year. Stenting was performed on large (>3.2 mm) coronary arteries for suboptimal results (47%), occlusive (8%), or nonocclusive dissections (45%) after balloon angioplasty. Stents were delivered using the bare stent technique and high pressure inflations (>12 atm). All patients received ticlopidine 250 mg (500 mg if weight was >80 kg) and aspirin 100 mg for 1 month. No patient received warfarin. At 1 week, 6 patients died of cardiogenic shock and 2 of right ventricular infarction. One subacute occlusion occurred at day 14. At 6 months, in 95 patients, the angiographic restenosis rate (>50% diameter stenosis) was 19%. One-year clinical follow-up, available in 55 patients, indicated cardiac death in 5, and repeat revascularization in 3. Thus, coronary stenting on large (>3.2 mm) coronary arteries after occlusive and nonocclusive dissection during primary balloon angioplasty for AMI using bare Palmaz-Schatz stents, high pressures, ticlopidine, and aspirin is safe. Our reocclusion and restenosis rates are similar to those of trials on elective stenting in stable patients. PMID- 9202347 TI - Intravascular ultrasonic evidence for importance of plaque distribution (eccentric vs circumferential) in determining distensibility of the left anterior descending artery. AB - Although previous studies have shown that coronary atherosclerosis is accompanied by impaired vessel wall compliance, few data exist regarding the regional vessel distensibility that may be important in order to gain an insight into the mechanism of atherosclerotic plaque rupture. Therefore, we analyzed 45 coronary sites of the proximal left anterior descending artery from 40 patients. Using intravascular ultrasound, luminal area in diastole (A) and in systole was measured at the diseased sites. With the ratio of luminal area changes (dA) to coronary pressure changes (dP) during a cardiac cycle, the total distensibility index was obtained by the formula: [(dA/A)/dP] x 10(3). At the sites with noncircumferential disease perimeters in diastole (L) and in systole were measured at the normal and narrowed portions. Using the changes in perimeters (dL) during a cardiac cycle, the regional distensibility index was obtained by the formula: [(dL/L)/dP] x 10(3). In 22 sites with circumferential disease, the total distensibility index was 1.03 +/- 0.61/mm Hg (mean +/- SD), and significantly lower than that from 23 sites with noncircumferential disease that showed 1.45 +/- 0.89/mm Hg (p <0.05). In noncircumferential disease, the regional distensibility index at narrowed portions was significantly lower, 0.33 +/- 0.47/mm Hg, than that at normal portion, 1.11 +/- 0.75/mm Hg (p <0.01), suggesting the heterogenous distribution of regional wall distensibility in noncircumferential lesions. These results indicate that the heterogeneous regional wall distensibility exists at the sites with noncircumferential disease where the total vessel distensibility is preserved by the presence of the compliant normal portion. PMID- 9202348 TI - Comparison of men versus women in cross-sectional area luminal narrowing, quantity of plaque, presence of calcium in plaque, and lumen location in coronary arteries by intravascular ultrasound in patients with stable angina pectoris. AB - Women have an increased mortality after coronary interventions compared with men, which may be partly explained by differences in comorbid clinical conditions. However, whether women also have quantitative differences in coronary atherosclerosis is not known. Preinterventional intravascular ultrasound (IVUS) was used to study de novo, nonostial native coronary lesions in 169 women and 549 men with chronic angina. The external elastic membrane (EEM), lumen, and plaque + media (P + M) areas, plaque burden, plaque eccentricity, and calcium were measured at the target lesion and at a proximal reference site. All cross sectional IVUS measures were also corrected for body surface area. Results are reported as mean +/- 1 SD. Women had significantly smaller reference site EEM (16.5 +/- 5.3 vs 19.4 +/- 6.3 mm2, p <0.0001), lumen (8.7 +/- 3.0 vs 9.9 +/- 4.0 mm2, p = 0.0020), and P + M areas (7.8 +/- 3.7 vs 9.5 +/- 4.2 mm2, p = 0.0001). Women also had significantly smaller lesion site EEM (16.2 +/- 5.9 vs 18.3 +/- 6.7 mm2, p = 0.0028), lumen (2.4 +/- 1.7 vs 2.9 +/- 2.6 mm2, p = 0.0273), and P + M areas (13.6 +/- 5.7 vs 15.3 +/- 6.4 mm2, p = 0.0112). However, when corrected for BSA, these differences were no longer significant. Women and men also had similar reference and lesion plaque burden, eccentricity, and calcium. Preinterventional IVUS analysis failed to detect any quantitative or qualitative differences in coronary atherosclerosis in men compared with women. PMID- 9202349 TI - Determinants of transplant-related coronary calcium detected by ultrafast computed tomography scanning. AB - Coronary calcium detected by ultrafast computed tomography (CT) has been shown to be a marker of coronary artery disease in heart transplant recipients. The objective of this study was to examine the possible determinants of coronary calcium after heart transplantation. Over a 15-month period, 102 consecutive cardiac transplant recipients (mean age 53 years, 88 men) underwent ultrafast CT scanning of the heart, in addition to coronary angiography, to determine coronary calcium score on their annual follow-up (a median of 4.6 years [range 63 days to 9.1 years] after transplant). The following data were also recorded: the recipient's sex and date of birth, date of transplantation, date of ultrafast computed tomography and coronary angiography; recipient pretransplant diagnosis, history of diabetes mellitus and systemic hypertension, fasting lipid profile, immunosuppression, number of rejection episodes, and donor organ ischemic time. Forty six patients (45.1%) had total calcium scores >0 and 41 (40.2%) had at least 1 major coronary with angiographic narrowing >24%. On univariate analysis, coronary calcium was significantly associated with dyslipoproteinemia, total cholesterol was >6.0 mmol/L (240 mg/dl), triglycerides were >3.0 mmol/L (265 mg/dl), and lipoprotein(a) >30 mg/ dl; > or =25% angiographic disease was significantly associated with coronary calcium and dyslipoproteinemia. Logistic regression revealed that dyslipoproteinemia, systemic hypertension, and donor ischemic time were significant predictors of coronary calcium in transplanted hearts. We conclude that the prevalence of coronary calcium in heart transplant recipients is high and is related to recipient dyslipoproteinemia, systemic hypertension, and donor organ ischemic time. PMID- 9202350 TI - Prognostic value of dobutamine stress echocardiography in patients undergoing diagnostic coronary arteriography. AB - There are only a few studies addressing the prognostic value of dobutamine stress echocardiography in patients with suspected coronary artery disease and none have assessed its value compared with coronary arteriography. Accordingly, graded dobutamine stress echocardiography was performed in 121 patients who underwent coronary arteriography based on symptoms and the findings of treadmill exercise electrocardiography. During the follow-up period of mean (SD) months (15 +/- 9) there were 41 cardiac events (death [n = 5], acute myocardial infarction [n = 2], unstable angina [n = 29], and congestive heart failure [n = 5]). There were a greater number of patients with inducible wall motion abnormality (88%) on dobutamine stress with cardiac events compared with those without (55%, p <0.001). The wall motion score indexes at rest (1.6 +/- 0.6) and at peak stress (2.1 +/- 0.8) were worse in patients with cardiac events compared with those without (1.2 +/- 0.3, p <0.001 and 1.5 +/- 0.6, p <0.001, respectively). When multivariate analysis was performed using clinical, exercise, echocardiographic, and coronary arteriographic data the independent predictors of cardiac events were exercise duration (p = 0.01), presence of inducible wall motion abnormality (p = 0.03), and wall motion score index at peak stress (p <0.001). Thus, dobutamine stress echocardiography is a powerful predictor of future cardiac events in patients undergoing exercise testing and coronary arteriography for evaluation of chest pain and is superior to both exercise electrocardiography and coronary arteriography for the prediction of subsequent cardiac events. PMID- 9202351 TI - Impact of impaired coronary flow reserve and insulin resistance on myocardial energy metabolism in patients with syndrome X. AB - To evaluate the role of a decreased coronary flow reserve in the genesis of angina pectoris in patients with syndrome X, we studied myocardial hemodynamics and metabolism at rest, during pace stress, and in the recovery period after pacing in 18 consecutive patients with syndrome X and in 10 control subjects. By means of positron emission tomography or the intracoronary flow-wire method, patients were subclassified as having microvascular angina (MA, n = 8) when coronary flow reserve was reduced (<2.5) or no microvascular angina (non-MA, n = 10) when coronary flow reserve was preserved (> or =2.5). At rest, coronary sinus blood flow was increased in MA patients. During pace stress, coronary sinus blood flow increased by 39 +/- 6% in MA patients versus 67 +/- 12% in non-MA patients and 69 +/- 7% in controls (p <0.05). Patients with non-MA revealed fasting hyperinsulinemia, increased arterial concentration of free fatty acids, and a similar tendency for beta-hydroxybutyrate. Oxygen extraction and carbon dioxide release did not differ between groups. Net myocardial lactate release was not observed in any patient during pace stress and myocardial energy metabolism was preserved in all patients with syndrome X. During pacing, myocardial uptake of free fatty acids and beta-hydroxybutyrate was increased in non-MA patients. Myocardial uptake of free fatty acids correlated positively and myocardial glucose and lactate uptake correlated inversely with arterial concentrations of free fatty acids in all subjects. Metabolic evidence of myocardial ischemia is uncommon in patients with syndrome X, irrespective of a globally reduced coronary flow reserve. Although patients with syndrome X can be subclassified according to presence of a microvascular or a metabolic disorder, angina pectoris and ST segment depressions coexist with a preserved global myocardial energy efficiency in all patients. PMID- 9202352 TI - Clinical predictors of transvenous biphasic defibrillation thresholds. AB - Transvenous lead systems have become routine for defibrillator placement. However, previous studies of clinical predictors of an adequate nonthoracotomy defibrillation threshold (DFT) evaluated monophasic waveforms or more complex lead systems, including subcutaneous patches. Accordingly, this study is a prospective evaluation of the predictors of an adequate biphasic DFT in 114 consecutive patients undergoing cardioverter-defibrillator implantation with a single transvenous lead. For each subject, 38 parameters were assessed, including standard demographic, electrocardiographic, echocardiographic, and radiographic measurements. An adequate DFT (< or =20 J) was achieved in 92% of patients. Multivariable analysis revealed 2 independent factors predictive of a high threshold: echocardiographic measurements of left ventricular dilation (odds ratio = 0.16, 95% confidence interval 0.05 to 0.53, p = 0.003) and body size (odds ratio = 0.36, 95% confidence interval 0.17 to 0.73; p = 0.005). No patient with a normal left ventricular end-diastolic dimension had a high DFT, whereas 14% (9 of 66) of those with left ventricular dilation had elevated thresholds. When the DFT cutoff was lowered to 15 J, as is necessary with some downsized pulse generators, an adequate threshold was observed in 84% of patients and the same 2 independent predictors of high thresholds were found. These results indicate that an adequate transvenous DFT can be predicted from simple clinical parameters. PMID- 9202353 TI - Influence of beta-adrenergic and vagal activity on the effect of exogenous adenosine on supraventricular tachycardia termination. AB - Adenosine, which binds to cell surface receptors and couples with guanosine triphosphate-binding inhibitory proteins (G(i)), is potent in terminating supraventricular tachycardia (SVT). However, whether the differences in autonomic tone will influence this effect remains unknown. This study was designed to investigate the role of beta-adrenergic and vagal activity on the action of adenosine. Forty patients with clinically documented SVT (22 with atrioventricular node reentrant tachycardia and 18 with atrioventricular reciprocating tachycardia) were divided into 4 groups with 10 patients in each group. In groups 1 and 2, adenosine was intravenously injected during the baseline state and during infusion of isoproterenol (2 and 4 microg/min, respectively). Group 2 patients received atropine (0.04 mg/kg) injection before isoproterenol infusion. In groups 3 and 4, intravenous injection of adenosine was given during the baseline state and after injection of atropine (0.02 and 0.04 mg/kg, respectively). Group 4 patients received propranolol (0.2 mg/kg) before atropine injection. The minimal dose of adenosine to terminate tachycardia during isoproterenol infusion of 2 microg/min was greater than that during the baseline state in both groups 1 and 2. The minimal dose of adenosine during isoproterenol infusion with 4 microg/min was higher than that with 2 microg/min in group 2, but not in group 1 patients. In both groups 3 and 4, the minimal dose of adenosine required to terminate tachycardia during atropine injection with 0.02 mg/kg was greater than that during the baseline state. The minimal effective dose of adenosine during atropine injection with 0.04 mg/kg was higher than that with 0.02 mg/kg in group 4, but not in group 3 patients. In conclusion, either limb of the autonomic nervous system may modulate the adenosine dosage required for termination of SVT. Patients taking drugs such as beta blockers or vagolytic agents may need alterations in the dose of adenosine for therapy. PMID- 9202354 TI - Low-energy endocardial defibrillation using dual, triple, and quadruple electrode systems. AB - The feasibility of achieving both universal application of nonthoracotomy leads and low (< or = 15 J) defibrillation energy requirements by optimizing lead system configuration for use with low-output (<30 J) biphasic shock pulse generators was examined. Sixteen patients (mean age 62 +/- 8 years and mean left ventricular ejection fraction of 38 +/- 15%) were included in the study. All patients had either experienced syncope with induced ventricular tachycardia (n = 4) or had documented sustained ventricular tachycardia (n = 7) or ventricular fibrillation (n = 5). Defibrillation threshold testing was performed in 2 stages on different days in these patients. In the first stage, 2 defibrillation catheter electrodes were positioned in the right ventricle and superior vena cava with an axillary cutaneous patch. Fifteen-joule, 10- and 5-J biphasic shocks were delivered across 3 different electrode configurations-right ventricle to superior vena cava, right ventricle to axillary patch, right ventricle to a combination of superior vena cava and axillary patch. In the second stage, an 80-ml can electrode was added subcutaneously in a pectoral location to the previous leads. Configurations compared were the right ventricle to pectoral can, and right ventricle to an "array"-combining superior vena cava, can, and axillary patch leads. The defibrillation threshold was determined using a step-down method. In stage 1, mean defibrillation threshold for the right ventricle to axillary patch (12.7 +/- 5.9 J) and right ventricle to superior vena cava plus axillary patch (9.8 +/- 5.2 J) configurations was lower than the right ventricle to superior vena cava configuration (14.2 +/- 6.4 J, p <0.05). In stage 2, the defibrillation was higher for the right ventricle to pectoral can (9.2 +/- 5.1 J) configuration compared with the right ventricle to the array (5.6 +/- 3.6 J, p < or =0.05). The right ventricle to array had the lowest defibrillation threshold, whereas the right ventricle to pectoral can was the best dual electrode system. Low-energy endocardial defibrillation (< or =10 J) was feasible in 72% of tested patients with > 1 electrode configuration at 10 J, whereas only 53% of successful patients could be reverted at >1 electrode configuration at 5 J (p <0.05). Reduction in maximum pulse generator output to < or =25 J using these electrode configurations with bidirectional shocks is feasible and maintains an adequate safety margin. PMID- 9202355 TI - Correlates of early hospital readmission or death in patients with congestive heart failure. AB - Among patients with heart failure who survive an admission to the hospital, those who are readmitted or die soon after discharge may warrant special attention. Therefore, we prospectively followed 257 patients admitted nonelectively to an urban university hospital, with a complaint of shortness of breath or fatigue and evidence of congestive heart failure on admission chest radiograph, who were discharged alive. Through survey of patients and families, review of the hospital computer system, and a search of the National Death Index, we recorded death and hospital readmission. Within 60 days of discharge, 13 patients (5%) died and 82 (32%) died or were readmitted to the hospital. Using Cox proportional-hazards modeling, the multivariable correlates of readmission or death were single marital status (adjusted hazard ratio [HR] 2.1, 95% confidence interval [CI] 1.3 to 3.3), Charlson Comorbidity Index score (HR 1.3 per point to maximum 4 points, 95% CI 1.1 to 1.6), admission systolic blood pressure of < or = 100 mm Hg (HR 2.8, 95% CI 1.6 to 5.0), and absence of new ST-T-wave changes on the initial electrocardiogram (HR 1.9, 95% CI 1.1 to 3.3). Self-reported patient compliance and clinical instability at discharge were not correlates. Almost all patients stratified by these factors had at least a 25% risk of readmission or death. Our independent correlates of readmission or death support the importance of both medical and social factors in the pathway to clinical decline. However, we could not reliably identify a truly low-risk group. Interventions to decrease early readmission or death among patients with heart failure should target both medical management and the adequacy of social support, and probably need to be applied to all admitted patients. PMID- 9202356 TI - Depressed heart rate variability as an independent predictor of death in chronic congestive heart failure secondary to ischemic or idiopathic dilated cardiomyopathy. AB - After acute myocardial infarction, depressed heart rate variability (HRV) has been proven to be a powerful independent predictor of a poor outcome. Although patients with chronic congestive heart failure (CHF) have also markedly impaired HRV, the prognostic value of HRV analysis in these patients remains unknown. The aim of this study was to investigate whether HRV parameters could predict survival in 102 consecutive patients with moderate to severe CHF (90 men, mean age 58 years, New York Heart Association [NYHA] class II to IV, CHF due to idiopathic dilated cardiomyopathy in 24 patients and ischemic heart disease in 78 patients, ejection fraction [EF], 26%; peak oxygen consumption, 16.9 ml/kg/min) after exclusion of patients in atrial fibrilation with diabetes or with chronic renal failure. In the prognostic analysis (Cox proportional-hazards model, Kaplan Meier survival analysis), the following factors were investigated: age, CHF etiology, NYHA class, EF, peak oxygen consumption, presence of ventricular tachycardia on Holter monitoring, and HRV measures derived from 24-hour electrocardiography monitoring, calculated in the time (standard deviation of all normal RR intervals [SDNN], standard deviation of 5-minute RR intervals [SDANN], mean of all 5-minute standard deviations of RR intervals [SD], root-mean-square of difference of successive RR intervals [rMSSD], and percentage of adjacent RR intervals >50 ms different [pNN50]) and frequency domain (total power [TP], power within low-frequency band [LF], and power within high-frequency band [HF]). During follow-up of 584 +/- 405 days (365 days in all who survived), 19 patients (19%) died (mean time to death: 307 +/- 315 days, range 3 to 989). Cox's univariate analysis identified the following factors to be predictors of death: NYHA (p = 0.003), peak oxygen consumption (p = 0.01), EF (p = 0.02), ventricular tachycardia on Holter monitoring (p = 0.05), and among HRV measures: SDNN (p = 0.004), SDANN (p = 0.003), SD (p = 0.02), and LF (p = 0.003). In multivariate analysis, HRV parameters (SDNN, SDANN, LF) were found to predict survival independently of NYHA functional class, EF, peak oxygen consumption, and ventricular tachycardia on Holter monitoring. The Kaplan-Meier survival curves revealed SDNN < 100 ms to be a useful risk factor; 1-year survival in patients with SDNN < 100 ms was 78% when compared with 95% in those with SDNN > 100 ms (p = 0.008). The coexistence of SDNN < 100 ms and a peak oxygen consumption < 14 ml/kg/min allowed identification of a group of 18 patients with a particularly poor prognosis (1-year survival 63% vs 94% in the remaining patients, p <0.001). We conclude that depressed HRV on 24-hour ambulatory electrocardiography monitoring is an independent risk factor for a poor prognosis in patients with CHF. Whether analysis of HRV could be recommended in the risk stratification for better management of patients with CHF needs further investigation. PMID- 9202357 TI - Comparison of morphologic assessment of hypertrophic cardiomyopathy by magnetic resonance versus echocardiographic imaging. AB - To compare the value of echocardiography and magnetic resonance imaging (MRI) in the assessment of the amount and extent of hypertrophy in hypertrophic cardiomyopathy (HC) and, second, to correlate the degree of hypertrophy, as assessed by MRI, with clinical and electrocardiographic parameters, 30 consecutive patients (16 men and 14 women, aged 20 to 74 years) with HC were studied. Measurements of left ventricular wall thickness were performed at 11 predetermined segments (5 basal, 5 midventricular, and 1 apical) by 2-dimensional echocardiography and MRI. Two parameters derived from MRI studies were considered as indicators of the degree and extent of hypertrophy: (1) mean of the measured wall thickness at the 11 segments, and (2) the number of segments with thickness > 15 mm. Results showed that, from a total of 330 myocardial segments, thickness could be measured by echocardiography in 221 (67%), whereas MRI allowed measurement of 320 segments (97%). When compared with clinical and electrocardiographic data, no correlation was found regarding mean wall thickness and number of hypertrophied segments by MRI except for the presence of an abnormal electrocardiographic repolarization pattern. It is concluded that MRI allows a better assessment of the degree and extension of left ventricular hypertrophy than echocardiography in HC. Despite the precise information on hypertrophy provided by MRI, the amount and degree of hypertrophy bears no correlation with most of the clinical data in these patients. PMID- 9202358 TI - Enhancement of left ventricular cavity opacification by harmonic imaging after venous injection of Albunex. AB - Venous injection of Albunex does not consistently produce left ventricular (LV) cavity opacification during conventional echocardiography. We postulated that by increasing the signal-to-noise ratio, harmonic imaging will result in more successful LV cavity opacification and provide a better assessment of regional LV systolic function. Forty-two patients with poor baseline endocardial delineation were given 10 ml intravenous injections of Albunex during continuous fundamental and harmonic imaging. Change in segmental wall-thickening scores and the confidence levels for these scores were assessed for 3 observers with various levels of experience. Compared with fundamental imaging, harmonic imaging significantly improved the success of LV cavity opacification (83% vs 62%, p <0.05). The background-subtracted video intensity within the central two thirds of the LV cavity increased threefold (from 10 +/- 15 to 31 +/- 29, p <0.05) with harmonic imaging. The spatial extent of opacification increased from 40% of the LV cavity during fundamental imaging to 65% with harmonic imaging (p <0.001). The confidence level for assessing regional LV systolic function improved (p <0.05) after contrast administration, particularly when observer experience was limited. We conclude that in patients with poor endocardial definition, injection of intravenous Albunex should be combined with harmonic imaging to improve LV cavity opacification. PMID- 9202359 TI - Results of recent large cholesterol-lowering trials and implications for clinical management. AB - Three recent large clinical trials-the West of Scotland Coronary Prevention Study, the Scandinavian Simvastatin Survival Study, and the Cholesterol and Recurrent Events Trial-have all confirmed that cholesterol lowering with HMG-CoA reductase inhibitors is safe and effective therapy to prevent an initial or recurrent coronary event in patients at high risk for coronary heart disease. However, a number of questions related to the treatment of lipid disorders and risk reduction for coronary heart disease remain, including the cholesterol concentration at which treatment would best be initiated, the optimal cholesterol reduction or goal to be attained, and the mechanisms by which HMG-CoA reductase inhibitors reduce the risk for clinical events. PMID- 9202360 TI - Relation of duration of ST reelevation at reperfusion and improvement of left ventricular function after successful primary angioplasty of the left anterior descending coronary artery in anterior wall acute myocardial infarction. AB - We conducted a prospective study to investigate the relation between ST reelevation during primary angioplasty and improvement in left ventricular function. The duration, not the occurrence, of ST reelevation at reperfusion was associated with improvement in left ventricular function in patients with anterior wall acute myocardial infarction successfully recanalized by primary angioplasty. PMID- 9202361 TI - Fish consumption, n-3 fatty acids in cell membranes, and heart rate variability in survivors of myocardial infarction with left ventricular dysfunction. AB - To elucidate a possible antiarrhythmic effect of long-chained n-3 polyunsaturated fatty acids, heart rate variability was assessed in 52 patients with a previous myocardial infarction and left ventricular dysfunction. The content of n-3 polyunsaturated fatty acids in platelets was closely associated with the patient's fish-consuming habits, and a significant positive correlation was observed between the n-3 fatty acid docosahexaenoic acid and heart rate variability. PMID- 9202362 TI - Intravascular ultrasound analysis of reduction in progression of coronary narrowing by treatment with pravastatin. AB - Serial intravascular ultrasound studies were performed to evaluate the effect of pravastatin on coronary atherosclerotic plaque. Administration of pravastatin reduced serum lipid levels and progression of coronary artery atherosclerotic plaque. PMID- 9202363 TI - Inhibition of nitric oxide synthesis during the cold pressor test in patients with coronary artery disease. AB - The effects of a cold pressor test during intracoronary infusions of L-NMMA and normal saline were studied in patients with chronic stable angina and in patients with normal coronary arteriograms. The cold pressor test during saline infusion caused significant dilation of proximal and distal segments in patients with normal coronary arteriograms, and this dilation was abolished by L-NMMA infusion; in patients with coronary disease the cold pressor test during saline caused constriction of the stenoses and distal segments and this constriction was augmented by L-NMMA infusion. PMID- 9202364 TI - Hospital cost of complications associated with coronary artery bypass graft surgery. AB - We identified 6,791 coronary artery bypass grafting (CABG) cases using the Massachusetts hospital discharge data to quantify the contribution of complications to the cost of a hospitalization for CABG. After adjusting for in hospital mortality and baseline clinical severity as other contributors to cost, the additional costs associated with complications were substantial. PMID- 9202365 TI - Relation of unstable angina and myocardial infarction to progression of coronary narrowing in patients undergoing coronary artery bypass grafting (utilizing an internal mammary artery). AB - We examined the relation of an ischemic syndrome with the progression of coronary disease early (<3 years) after multiple bypass grafting utilizing an internal mammary artery and saphenous vein grafts. Data indicate that an ischemic syndrome is associated with progression of native coronary disease distal to the graft or total occlusion of the saphenous vein graft in most cases. PMID- 9202366 TI - Effects of nicorandil, an antianginal potassium channel opener, on left ventricular systolic and diastolic function in patients with chronic coronary artery disease. AB - We compared the effects of nicorandil with nitroglycerin and nifedipine on left ventricular function. Intravenous nicorandil may be a balanced-typed vasodilator and useful in patients with left ventricular systolic and diastolic dysfunction. PMID- 9202367 TI - Influence of cardiovascular risk status on coronary flow reserve in healthy young men. AB - To detect changes in vascular physiology associated with early atherosclerosis, we studied whether alterations in coronary flow reserve, as assessed by positron emission tomography imaging with intravenous dipyridamole, would be related to risk factor variables in healthy young men. The number of conventional risk variables correlated significantly with coronary flow reserve (r = -0.58, p = 0.0007), suggesting that alterations in functional vascular reactivity are related to the cardiovascular risk status already in healthy young men. PMID- 9202368 TI - Hormonal responses during tilt-table test in neurally mediated syncope. AB - The hormonal profile during tilt testing was examined in syncopal patients. An increase in the growth hormones cortisol and prolactin was found during syncope, suggesting an implication of central serotonergic activation. PMID- 9202369 TI - Comparison of early target organ damage between blacks and whites with mild systemic arterial hypertension. AB - As greater mortality and morbidity from target organ damage in arterial hypertension have been reported for black than for white hypertensives, we examined in a matched-pair analysis whether race per se affected markers of early target organ damage at similar levels of blood pressure. After controlling for the confounding factors such as age, sex, weight, and arterial pressure that interact with hypertension-related target organ damage, no racial disparities could be detected between matched black and white hypertensive patients. PMID- 9202370 TI - Serum levels of soluble form of Fas molecule in patients with congestive heart failure. AB - Compared with soluble Fas molecule (sFas, an inhibitor of Fas-mediated apoptosis) in normal volunteers, the serum level of sFas significantly increased by 41% in New York Heart Association (NYHA) class III (p <0.05) and by 97% in NYHA class IV patients with congestive heart failure (p <0.001). Furthermore, sFas showed correlations with soluble forms of TNF receptor-p55 (RI) and -p75 (RII) (r = 0.68 and r = 0.56) which inhibit activities of TNF alpha. PMID- 9202371 TI - Delayed recovery of postexercise blood pressure in patients with chronic heart failure. AB - Thirty-four patients with idiopathic dilated and ischemic cardiomyopathy underwent a symptom-limited cardiopulmonary exercise testing to evaluate the significance of postexercise blood pressure (BP) response. The postexercise BP response was useful in assessing the impaired exercise capacity and increased sympathetic activity in patients with heart failure. PMID- 9202372 TI - Transient ST-segment changes associated with mitral valvuloplasty using the Inoue balloon. AB - We describe unexplained transient inferior ST-segment elevation on the electrocardiogram during Inoue mitral valvuloplasty in 8 patients from a series of 108. Electrocardiographic changes were associated with chest pain in 7 patients, and although the clinical features were suggestive of myocardial ischemia, no cause for this could be found. PMID- 9202373 TI - Pericardial effusion in adults undergoing surgical repair of atrial septal defect. AB - The incidence of pericardial effusion and tamponade postatrial septal defect repair in adult patients are 16 and 1.5%, respectively. Small, medium, and large effusions progressed equally, and echocardiographic study on days 7, 14, and 28 best detects potentially significant effusion. PMID- 9202374 TI - Characteristics of 161 patients with cardiac tumors diagnosed during 1993 and 1994 in Japan. AB - We investigated clinical and pathologic characteristics of 161 patients with primary or secondary cardiac tumors diagnosed between 1993 and 1994 in Japan. The increased use of cardiovascular imaging, especially echocardiography, contributed to the early identification of small cardiac tumors, resulting in a reduction of the serious complications such as embolization. PMID- 9202375 TI - Possible association of thrombotic, nonbacterial vegetations of the mitral ring mitral annular calcium and stroke. AB - Although numerous studies have shown an increased risk of stroke associated with mitral annular calcification, a direct link has rarely been demonstrated. We report the occurrence of long, pedunculated thrombi attached to the calcified mitral annulus in 3 patients who suffered from stroke, with resolution after anticoagulant and antithrombotic therapy. PMID- 9202377 TI - Construction of an 800-kb contig in the near-centromeric region of the rice blast resistance gene Pi-ta2 using a highly representative rice BAC library. AB - We constructed a rice Bacterial Artificial Chromosome (BAC) library from green leaf protoplasts of the cultivar Shimokita harboring the rice blast resistance gene Pi-ta. The average insert size of 155 kb and the library size of seven genome equivalents make it one of the most comprehensive BAC libraries available, and larger than many plant YAC libraries. The library clones were plated on seven high density membranes of microplate size, enabling efficient colony identification in colony hybridization experiments. Seven percent of clones carried chloroplast DNA. By probing with markers close to the blast resistance genes Pi-ta2(closely linked to Pi-ta) and Pi-b, respectively located in the centromeric region of chromosome 12 and near the telomeric end of chromosome 2, on average 2.2 +/- 1.3 and 8.0 +/- 2.6 BAC clones/marker were isolated. Differences in chromosomal structures may contribute to this wide variation in yield. A contig of about 800 kb, consisting of 19 clones, was constructed in the Pi-ta2 region. This region had a high frequency of repetitive sequences. To circumvent this difficulty, we devised a "two-step walking" method. The contig spanned a 300 kb region between markers located at 0 cM and 0.3 cM from Pi-ta. The ratio of physical to genetic distances (> 1,000 kb/cM) was more than three times larger than the average of rice (300 kb/cM). The low recombination rate and high frequency of repetitive sequences may also be related to the near centromeric character of this region. Fluorescent in situ hybridization (FISH) with a BAC clone from the Pi-b region yielded very clear signals on the long arm of chromosome 2, while a clone from the Pi-ta2 region showed various cross hybridizing signals near the centromeric regions of all chromosomes. PMID- 9202378 TI - A trans-acting modifier causing extensive overexpression of genes in Drosophila melanogaster. AB - A modified P element-induced mutation at cytological position 47A11-12 on chromosome 2 of Drosophila melanogaster inversely alters the expression of several genes examined. In addition, the effect is nearly exponential in some developmental stages for five of the eleven genes tested (white, brown, scarlet, rudimentary and vermilion). Interestingly, the exponential overexpression of the transcripts leads to a sexually dimorphic effect, with female levels being greater. The white allele specificity analysis and the lack of response of an Alcohol dehydrogenase promoter-white reporter fusion construct suggest that the 5' regulatory sequences of white are required for the response. Because the cases of extreme overexpression are more prominent in females than males, the locus is termed Ultra female overexpression (Ufo). Ufo revertants have no discernible effect on the transcript levels, indicating that the mutation is responsible for the modulation of the tested genes. Regulatory genes such as Ufo may contribute to the wide fluctuations in gene expression between tissues and stages of development. PMID- 9202379 TI - Gene cloning, sequencing and enzymatic properties of glutamate synthase from the hyperthermophilic archaeon Pyrococcus sp. KOD1. AB - The gltA gene encoding a glutamate synthase (GOGAT) from the hyperthermophilic archaeon Pyrococcus sp. KOD1 was cloned as a 6.6 kb HindIII-BamHI fragment. Sequence analysis indicates that gltA encodes a 481- amino acid protein (53,269 Da). The deduced amino acid sequence of KOD1-GltA includes conserved regions that are found in the small subunits of bacterial GOGAT: two cysteine clusters, an adenylate-binding consensus sequence and an FAD-binding consensus sequence. However, no sequences homologous to the large subunit of bacterial GOGAT were found in the upstream or downstream regions. In order to examine whether GltA alone can act as a functional GOGAT, GltA was overexpressed in Escherichia coli BL21 (DE3) cells using an expression plasmid. GltA was purified to homogeneity and shown to be functional as a homotetramer of approximately 205 kDa, which is equivalent to the molecular weight of the native GOGAT from KOD1, thus indicating that KOD1-GOGAT is the smallest known active GOGAT. GltA is capable of both glutamine-dependent and ammonia-dependent synthesis of glutamate. Synthesis of glutamate by KOD1-GltA required NADPH, indicating that this enzyme is an NADPH GOGAT (EC 1.4.1.13). The optimum pH for both activities was 6.5. However, GltA exhibited different optimum temperatures for activity depending on the reaction assayed (glutamine-dependent reaction, 80 degrees C; ammonia-dependent reaction, 90 degrees C). PMID- 9202380 TI - Two uvs genes of Aspergillus nidulans with different functions in error-prone repair: uvsI, active in mutation-specific reversion, and uvsC, a recA homolog, required for all UV mutagenesis. AB - Two genes of Aspergillus nidulans are known to function in UV mutagenesis, but have been assigned to different epistasis groups: uvsC, which is also required for meiosis and mitotic recombination, and uvsI, which may have no other function. To clarify their role in error-prone repair and to investigate their interaction, uvsI and uvsC single and uvsI;uvsC double mutant strains were further tested for mutagen sensitivities and characterized for effects on mutation. Spontaneous and induced frequencies were compared in forward and reverse mutation assays. All results confirmed that uvsI and uvsC are members of different epistasis groups, and demonstrated that these uvs mutants have very different defects in UV mutagenesis. The uvsI strains showed wild-type frequencies in all forward mutation tests, but greatly reduced spontaneous and UV induced reversion of some, but not other, point mutations. In contrast, uvsC had similar effects in all assay systems: namely pronounced mutator effects and greatly reduced UV mutagenesis. Interestingly, the uvsI;uvsC double mutant strains differed from both single mutants; they clearly showed synergism for all types of reversion tested: none were ever obtained spontaneously, nor after induction by UV or EMS (ethylmethane sulfonate). Based on these results, we conclude that uvsI is active in a mutation-specific, specialized error-prone repair process in Aspergillus. In contrast, uvsC, which is now known to show sequence homology to recA, has a basic function in mutagenic UV repair in addition to recombinational repair, similar to recA of Escherichia coli. PMID- 9202381 TI - Cloning, sequencing, disruption and phenotypic analysis of uvsC, an Aspergillus nidulans homologue of yeast RAD51. AB - We have cloned the uvsC gene of Aspergillus nidulans by complementation of the A. nidulans uvsC114 mutant. The predicted protein UVSC shows 67.4% sequence identity to the Saccharomyces cerevisiae Rad51 protein and 27.4% sequence identity to the Escherichia coli RecA protein. Transcription of uvsC is induced by methyl-methane sulphonate (MMS), as is transcription of RAD51 of yeast. Similar levels of uvsC transcription were observed after MMS induction in a uvsC+ strain and the uvsC114 mutant. The coding sequence of the uvsC114 allele has a deletion of 6 bp, which results in deletion of two amino acids and replacement of one amino acid in the translation product. In order to gain more insight into the biological function of the uvsC gene, a uvsC null mutant was constructed, in which the entire uvsC coding sequence was replaced by a selectable marker gene. Meiotic and mitotic phenotypes of a uvsC+ strain, the uvsC114 mutant and the uvsC null mutant were compared. The uvsC null mutant was more sensitive to both UV and MMS than the uvsC114 mutant. The uvsC114 mutant arrested in meiotic prophase-I. The uvsC null mutant arrested at an earlier stage, before the onset of meiosis. One possible interpretation of these meiotic phenotypes is that the A. nidulans homologue of Rad51 of yeast has a role both in the specialized processes preceding meiosis and in meiotic prophase I. PMID- 9202383 TI - Flipper, a mobile Fot1-like transposable element in Botrytis cinerea. AB - A transposable element, Flipper, was isolated from the phytopathogenic fungus Botrytis cinerea. The element was identified as an insertion sequence within the coding region of the nitrate reductase gene. The Flipper sequence is 1842 bp long with perfect inverted terminal repeats (ITRs) of 48 bp and an open reading frame (ORF) of 533 amino acids, potentially encoding for a transposase; the element is flanked by the dinucleotide TA. The encoded protein is very similar to the putative transposases of three elements from other phytopathogenic fungi, Fot1 from Fusarium oxysporum, and Pot2 and MGR586 from Magnaporthe grisea. The number of Flipper elements in strains of B. cinerea varied from 0 to 20 copies per genome. Analysis of the descendants of one cross showed that the segregation ratio of Flipper elements was 2:2 and that the copies were not linked. PMID- 9202382 TI - Genetic evidence for 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) as a negative effector of cytochrome terminal oxidase cbb3 production in Rhizobium etli. AB - A Rhizobium etli Tn5mob-induced mutant (CFN035) exhibits an enhanced capacity to oxidize N,N,N',N', tetramethyl-p -phenylenediamine (TMPD), a presumptive indicator of elevated cytochrome c terminal oxidase activity. Sequencing of the mutated gene in CFN035 revealed that it codes for the amidophosphoribosyl transferase enzyme (PurF) that catalyzes the first step in the purine biosynthetic pathway. Two c-type cytochromes with molecular weights of 32 and 27 kDa were produced in strain CFN035, which also produced a novel CO-reactive cytochrome (absorbance trough at 553 nm), in contrast to strain CE3 which produced a single 32 kDa c-type protein and did not produce the 553 nm CO reactive cytochrome. A wild-type R. etli strain that expresses the Bradyrhizobium japonicum fixNOQP genes, which code for the symbiotic cytochrome terminal oxidase cbb3, produced similar absorbance spectra (a trough at 553 nm in CO-difference spectra) and two c-type proteins similar in size to those of strain CFN035, suggesting that CFN035 also produces the cbb3 terminal oxidase. The expression of a R. etli fixN-lacZ gene fusion was measured in several R. etli mutants affected in different steps of the purine biosynthetic pathway. Our analysis showed that purF, purD, purQ, purL, purY, purK and purE mutants expressed three-fold higher levels of the fixNOQP operon than the wild-type strain. The derepressed expression of fixN was not observed in a purH mutant. The purH gene product catalyzes the conversion of 5-aminoimidazole-4-carboxamide ribonucleotide (AICAR) to 5-form-aminoimidazole-4-carboxamide ribonucleotide (FAICAR) and inosine. Supplementation with AICA riboside lowered the levels of fixN expression in the purF mutants. These data are consistent with the possibility that AICAR, or a closely related metabolite, is a negative effector of the production of the symbiotic terminal oxidase cbb3 in R. etli. PMID- 9202384 TI - A simple signal element mediates transcription termination and mRNA 3' end formation in the DEG1 gene of Saccharomyces cerevisiae. AB - DEG1 is a weakly transcribed gene of Saccharomyces cerevisiae, closely associated with CEN6. We mapped its major poly(A) site only 24 nucleotides (nt) downstream of the stop codon, and only 26 nt upstream of the CDEI centromere element. The deletion of this 50 nt stretch completely abolishes formation of the mRNA 3' end. A shorter deletion of a 16 nt sequence in the 3'-untranslated region has the same effect on transcription termination and 3'-maturation function. A TATATA sequence within this 16 nt region is essential for both functions, while a TGTATA sequence has a weak compensating activity in 3' end maturation if the TATATA stretch is deleted. We assume that the 3' end formation signals of the DEG1 gene have this simple structure: a single essential element (TATATA, whether alone or with the few surrounding nucleotides), probably, but not necessarily, cooperating with the sequence at the poly(A) site. This simple structure differs from the emerging model for 3' end-processing signals in that (i) it is shorter: 24 nt long at the most, while the model suggests 39 nt; (ii) there is no element located downstream of the TATATA signal to position the poly(A) site; and (iii) unlike the other naturally occurring signals studied, no cooperation among multiple TATATA-like elements is observed. We found that the same TATATA sequence also directs transcription termination, irrespective of promoter strength, and presumably without the cooperation of a downstream polymerase II pausing site. Taken together, these findings support the hypothesis that the DEG1 3' end-forming signals are more condensed than in other yeast genes, probably because of their proximity to CEN6. PMID- 9202385 TI - A helix-turn-helix DNA-binding motif predicted for transposases of DNA transposons. AB - A helix-turn-helix (HTH) DNA-binding motif is identified in transposase sequences in Tc1, mariner and pogo DNA transposum. The findings are supported by results of various sequence analysis methods. Tc1 transposases are also predicted to contain another DNA-binding region. These findings are in accord with experimental evidence obtained from Tc1A, Tc3A and pogo transposases. The pogo family transposases, but not the pogo-type transcription factors, contain the HTH motif, suggesting that HTH structures are essential for Tc1/mariner/pogo transposition. Analysis of multiple sequence alignments enabled the identification of the HTH motif in distantly related protein sequences. PMID- 9202386 TI - Fibroblast growth factor receptor signalling has a role in lobuloalveolar development of the mammary gland. AB - We have used the mouse mammary tumor virus promoter to express two dominant negative (DN) fibroblast growth factor receptor (FGFR) isoforms in the mammary epithelium of transgenic mice. While expression of DN-FGFR1(IIIc) showed no discernible phenotype, a similar kinase negative form of FGFR2(IIIb) caused a marked impairment of lobuloalveolar development. The growth retardation was apparent by mid-pregnancy and persisted in the post-partum glands. Despite the substantial underdevelopment of the mammary gland there was a measurable lactational response, but it was insufficient to properly sustain the new-born pups. These findings demonstrate that fibroblast growth factor signalling is necessary for pregnancy dependent lobuloalveolar development of the mammary gland. PMID- 9202387 TI - Tip localized Ca2+ pulses are coincident with peak pulsatile growth rates in pollen tubes of Lilium longiflorum. AB - It is known that locally elevated Ca2+ at the growing tips of pollen tubes is necessary for pollen tube elongation. Here we show that this localized Ca2+ is also temporally regulated and is closely associated with pulsatile tip growth. Lilium longiflorum pollen tubes were injected with the photoprotein, aequorin, and the Ca2(+)-dependent light output was detected with a low noise photon counting system. Ca2+ pulses with a mean period of 40 seconds were invariably associated with growth. The pulses were sporadic and of low amplitude for about the first 1.5 hours after germination. With subsequent growth, pulses increased in amplitude and the period between pulses became more regular. We have localized these Ca2+ pulses to the elongating end of the growing tube. The Ca2+ pulses are asymmetrical, rising more slowly than they fall. We estimate that the Ca2+ concentration at the peak of the pulses reaches nearly 10 microM. The addition of 100 microM La3+, a Ca2+ channel blocker, extinguished the pulses. An analysis of growth of elongating tubes establishes that extension is pulsatile, with a 42 second period between pulses. Calcium imaging, using the fluorescent indicator, Calcium Green dextran, shows that calcium pulses are coincident with peak growth rates. PMID- 9202388 TI - Differentiation of embryonic stem cells into adipocytes in vitro. AB - Embryonic stem cells, derived from the inner cell mass of murine blastocysts, can be maintained in a totipotent state in vitro. In appropriate conditions embryonic stem cells have been shown to differentiate in vitro into various derivatives of all three primary germ layers. We describe in this paper conditions to induce differentiation of embryonic stem cells reliably and at high efficiency into adipocytes. A prerequisite is to treat early developing embryonic stem cell derived embryoid bodies with retinoic acid for a precise period of time. Retinoic acid could not be substituted by adipogenic hormones nor by potent activators of peroxisome proliferator-activated receptors. Treatment with retinoic acid resulted in the subsequent appearance of large clusters of mature adipocytes in embryoid body outgrowths. Lipogenic and lipolytic activities as well as high level expression of adipocyte specific genes could be detected in these cultures. Analysis of expression of potential adipogenic genes, such as peroxisome proliferator-activated receptors gamma and delta and CCAAT/enhancer binding protein beta, during differentiation of retinoic acid-treated embryoid bodies has been performed. The temporal pattern of expression of genes encoding these nuclear factors resembled that found during mouse embryogenesis. The differentiation of embryonic stem cells into adipocytes will provide an invaluable model for the characterisation of the role of genes expressed during the adipocyte development programme and for the identification of new adipogenic regulatory genes. PMID- 9202389 TI - Phosphorylation regulates the assembly of NuMA in a mammalian mitotic extract. AB - NuMA is a 236 kDa nuclear protein that is required for the organization of the mitotic spindle. To determine how NuMA redistributes in the cell during mitosis, we have examined the behavior of NuMA in a mammalian mitotic extract under conditions conducive to the reassembly of interphase nuclei. NuMA is a soluble protein in mitotic extracts prepared from synchronized cultured cells, but forms insoluble structures when the extract becomes non-mitotic (as judged by the inactivation of cdc2/cyclin B kinase and the disappearance of mpm-2-reactive antigens). These NuMA-containing structures are irregularly shaped particles of 1 2 microm in diameter and their assembly is specific because other nuclear components such as the lamins remain soluble in the extract under these conditions. NuMA is dephosphorylated during this assembly process, and the assembly of these NuMA-containing structures is catalyzed by protein dephosphorylation because protein kinase inhibitors enhance their formation and protein phosphatase inhibitors block their formation. Finally, immunodepletion demonstrates that NuMA is an essential structural component of these insoluble particles, and electron microscopy shows that the particles are composed of a complex interconnected network of foci. These results demonstrate that phosphorylation regulates the solubility of NuMA in a mammalian mitotic extract, and the spontaneous assembly of NuMA into extensive structures upon dephosphorylation supports the conclusion that NuMA serves a structural function. PMID- 9202390 TI - Vam3p, a new member of syntaxin related protein, is required for vacuolar assembly in the yeast Saccharomyces cerevisiae. AB - Syntaxins are thought to participate in the specific interactions between vesicles and acceptor membranes in intracellular protein trafficking. VAM3 of Saccharomyces cerevisiae encodes a 33 kDa protein (Vam3p) with a hydrophobic transmembrane segment at its C terminus. Vam3p has structural similarities to syntaxins of yeast, animal and plant cells. delta vam3 cells accumulated spherical structures of 200-600 nm in diameter, but lacked normal large vacuolar compartments. Loss of function of Vam3p resulted in inefficient processing of vacuolar proteins proteinase A, proteinase B and carboxypeptidase Y, and defective maturation of alkaline phosphatase. Subcellular fractionation and immunofluorescence microscopy showed that Vam3p was localized to the vacuolar membranes. Vam3p was accumulated in certain regions of the vacuolar membranes. We conclude from these observations that Vam3p is a novel member of syntaxin in the vacuoles and it provides the t-SNARE function in a late step of the vacuolar assembly. PMID- 9202391 TI - Polarisation-dependent association of plectin with desmoplakin and the lateral submembrane skeleton in MDCK cells. AB - The intermediate filament-binding protein plectin and cytokeratin were localised at the cellular periphery of fully polarised Madin-Darby canine kidney (MDCK) cells, whereas vimentin was primarily found in a perinuclear network. Confocal and immunoelectron microscopy revealed that plectin was restricted to areas underlying the lateral plasma membrane. It colocalised with fodrin, a component of the submembrane skeleton, and was closely associated with desmosomal plaque structures. Biochemically, plectin was shown to interact directly with immunoprecipitated desmoplakin in vitro. Upon loss of cell polarity in low calcium medium, plectin redistributed to a cytoplasmic vimentin- and cytokeratin related network, clearly distinct from diffusely distributed fodrin and internalised desmoplakin structures. The structural reorganisation of plectin was also reflected by an increased solubility of the protein in Triton X-100/high salt, and a decrease in its half-life from approximately 20 to approximately 5 hours. Furthermore, unlike cytokeratins and vimentin, desmoplakin and fodrin did not associate with plectin attached to magnetic beads in cell lysates of unpolarised cells, while all proteins formed a stable complex in polarised cells. Altogether, these data indicate that plectin is involved in the anchorage of intermediate filaments to desmosomes and to the submembrane skeleton in polarised MDCK cells. PMID- 9202392 TI - Changing intestinal connective tissue interactions alters homeobox gene expression in epithelial cells. AB - In segmented organs, homeobox genes are involved in axial patterning and cell identity. Much less is known about their role in non-segmented endoderm derivatives such as the digestive epithelium. Using a xenograft model of fetal intestinal anlagen implanted under the skin of nude mice, we have investigated whether the expression of five homeobox genes (HoxA-4, HoxA-9, HoxC-8, Cdx-1 and Cdx-2) is modified when intestinal epithelium undergoes normal development or displays heterodifferentiation in association with heterotopic mesenchyme. In homotypic associations of fetal endoderm and mesenchyme that recapitulate normal development, the overall pattern of homeobox gene expression was maintained: HoxA 9 and HoxC-8 were the highest in the colon and ileum, respectively, and HoxA-4 was expressed all along the intestine; Cdx-1 and Cdx-2 exhibited an increasing gradient of expression from small intestine to colon. Yet, grafting per se caused a faint upregulation of HoxA-9 and HoxC-8 in small intestinal regions in which these genes are not normally expressed, while the endoderm-mesenchyme dissociation-association step provoked a decay of Cdx-1 in the colon. In heterotopic associations of colonic endoderm with small intestinal mesenchyme, the colonic epithelium exhibited heterodifferentiation to a small intestinal-like phenotype. In this case, we observed a decay of HoxA-9 expression and an upregulation of HoxC-8. Additionally, heterodifferentiation of the colonic epithelium was accompanied by a downregulation of Cdx-1 and Cdx-2 to a level similar to that found in the normal small intestine. To demonstrate that mesenchyme-derived cells can influence Cdx-1 and Cdx-2 expression in the bowel epithelium, fetal jejunal endoderm was associated with intestinal fibroblastic cell lines that either support small intestinal-like or colonic-like morphogenesis. A lower expression of both homeobox genes was shown in grafts presenting the small intestinal phenotype than in those showing glandular colonic like differentiation. Taken together, these results suggest that homeobox genes participate in the control of the positional information and/or cell differentiation in the intestinal epithelium. They also indicate that the level of Cdx-1 and Cdx-2 homeobox gene expression is influenced by epithelial mesenchymal cell interactions in the intestinal mucosa. PMID- 9202393 TI - Nuclear import of hnRNP A1 is mediated by a novel cellular cofactor related to karyopherin-beta. AB - Heterogeneous nuclear ribonucleoprotein A1 contains a sequence, termed M9, that functions as a potent nuclear localization signal (NLS) yet bears no similarity to the well-defined basic class of NLSs. Here, we report the identification of a novel human protein, termed MIP, that binds M9 specifically both in vivo and in vitro yet fails to interact with non-functional M9 point mutants. Of note, the 101 kDa MIP protein bears significant homology to human karyopherin/importin beta, a protein known to mediate the function of basic NLSs. The in vitro nuclear import of a protein substrate containing the M9 NLS was found to be dependent on provision of the MIP protein in trans. Cytoplasmic microinjection of a truncated form of MIP that retains the M9 binding site blocked the in vivo nuclear import of a substrate containing the M9 NLS yet failed to affect the import of a similar substrate bearing a basic NLS. These data indicate that nuclear import of hnRNP A1 is mediated by a novel cellular import pathway that is distinct from, yet evolutionarily related to, the pathway utilized by basic NLS sequences. PMID- 9202394 TI - Pathogen-induced programmed cell death in tobacco. AB - Sacrificing an infected cell or cells in order to prevent systemic spread of a pathogen appears to be a conserved strategy in both plants and animals. We studied some of the morphological and biochemical events that accompany programmed cell death during the hypersensitive response of tobacco plants infected with tobacco mosaic virus. Certain aspects of this cell death process appeared to be similar to those that take place during apoptosis in animal cells. These included condensation and vacuolization of the cytoplasm and cleavage of nuclear DNA to 50 kb fragments. In contrast, internucleosomal fragmentation, condensation of chromatin at the nuclear periphery and apoptotic bodies were not observed in tobacco plants during tobacco mosaic virus-induced hypersensitive response. A unique aspect of programmed cell death during the hypersensitive response of tobacco to tobacco mosaic virus involved an increase in the amount of monomeric chloroplast DNA. Morphological changes to the chloroplast and cytosol of tobacco cells and increase in monomeric chloroplast DNA occurred prior to gross changes in nuclear morphology and significant chromatin cleavage. Our findings suggest that certain aspects of programmed cell death may have been conserved during the evolution of plants and animals. PMID- 9202395 TI - Distribution of insulin/insulin-like growth factor-I hybrid receptors in human tissues. AB - Insulin receptors (IR) and type 1 IGF receptors (IGF-IR) have been shown to form insulin/IGF-I hybrid receptors in tissues expressing both molecules. The biological function of hybrid receptors is still undefined. To date there is no information about the distribution of hybrid receptors in human tissues. We have applied two microwell-based immunoassays which are capable of quantitating hybrid receptors in small samples of human tissues and cells. Results demonstrated that the proportion of total IGF-IR assembled as hybrids varied between 40 and 60%, thus indicating that hybrid receptors account for a large fraction of total IGF-I binding in human tissues. A significant fraction of total IR was assembled as hybrids in the tissues examined, varying from 37% in placenta to 45% in hepatoma, with the exception of adipose tissue where the fraction of insulin receptors forming hybrids was 17%. Because hybrid receptors bind IGF-I, but not insulin, with high affinity, it is likely that in human tissues hybrid receptors may be primarily activated by IGF-I rather than insulin under physiological conditions. Therefore, differences in hybrid receptors distribution may contribute to regulate tissue sensitivity to insulin and IGF-I by sequestering insulin receptor alphabeta-heterodimer in an IGF-I responsive form. PMID- 9202396 TI - Influence of age on 1,25(OH)2-vitamin D3 activation of protein kinase C in rat duodenum. AB - We have studied age-related changes in the non-genomic regulation of protein kinase C (PKC) by 1,25-dihydroxy-vitamin D3 [1,25(OH)2D3] and their role in 1,25(OH)2D3-dependent calcium uptake in the rat duodenum. Treatment of duodenal mucosae from 3 month-old (young) rats with hormone physiological concentrations (0.1 nM) induced an acute and transient stimulation of total tissue PKC activity which was maximal at 1 min (+80%). The responses were evidenced up to 10 nM 1,25(OH)2D3. The duodenum from 22 to 24 month old (aged) rats exhibited higher basal PKC activity which was not significantly modified after addition of the hormone. In the young duodenum PKC activation by 1,25(OH)2D3 was dependent on extracellular Ca2+ influx as it could be abolished to a great extent by EGTA and the Ca2+ channel blocker verapamil. In addition, the Ca2+ ionophore A23187 elicited a marked stimulation of duodenal mucosae PKC in young rats but was without effects in aged animals. 1,25(OH)2D3 increased the influx of 45Ca2+ in duodenal mucosae of young rats in a dose-(0.1-1 nM) and time-(1-10 min) dependent fashion. This response to the hormone was impaired in aged animals. Similarly as 1,25(OH)2D3, the PKC activator dioctanoylglycerol (DOG) rapidly (1-5 min) increased [45Ca2+] influx in duodena from young rats whereas the response to DOG was blunted in senescent animals. Furthermore, PKC inhibitors (bisindolylmaleimide, staurosporine and compound H7) abolished 1,25(OH)2D3 stimulation of Ca2+ uptake in the young duodenum. These results suggest that 1,25(OH)2D3 regulates PKC activity in the mammalian duodenum by a non-genomic mechanism which involves the rapid influx of extracellular Ca2+, and that activation of PKC, in turn, mediates hormone stimulation of intestinal Ca2+ uptake. The data also indicates that 1,25(OH)2D3 regulation of Ca2+ transport through the PKC messenger system is impaired with aging. PMID- 9202397 TI - Growth hormone down-regulates growth hormone receptor mRNA in chickens but developmental increases in growth hormone receptor mRNA occur independently of growth hormone action. AB - The purpose of this study was to determine the role of growth hormone (GH) in regulating expression of the chicken GH receptor (cGHR) gene by comparing the levels of cGHR mRNA in livers of normal chickens with that of GHR-deficient dwarf chickens. Since the sex-linked dwarf chicken lacks a functional cGHR, there are no genes activated as a result of GH action. Examination of the early developmental profile of hepatic cGHR mRNA in normal and dwarf chickens should yield information on the relative contribution of developmental and hormonal factors to the regulation of cGHR gene expression. Using a sensitive RNase protection assay, we found that the abundance of the major cGHR transcripts (4.3, 3.2 and 0.8 kb) in normal chickens increases about 2-fold between 1 and 7 weeks of age. Due to a splice site mutation in the dwarf chicken, the two larger transcripts encoding the full-length cGHR are not expressed. However, the expression of the truncated cGHR transcript (0.8 kb) in dwarf chickens increases about 5-fold between 1 and 7 weeks of age which suggests that the cGHR gene is overexpressed when not down-regulated by GH. Furthermore, a single promoter, appears to control expression of cGHR transcripts in liver since primer extension analysis revealed the same 5'-end in both full-length and 0.8 kb transcripts. These observations suggest that even though developmental increases in cGHR gene expression occur independently of GH action, GH, either directly or indirectly, down-regulates expression of the cGHR gene in normal chickens. PMID- 9202398 TI - Prolactin, epidermal growth factor or transforming growth factor-alpha activate a mammary cell-specific enhancer in mouse mammary tumor virus-long terminal repeat. AB - Mammary specific expression of elevated levels of mouse mammary tumor virus (MMTV) contributes to mammary carcinogenesis. Mechanisms which regulate provirus expression have not been completely defined. Using a MMTV-long repeat terminal (MMTV-LRT) directed chloramphenicol-acetyltransferase (CAT) reporter gene system and a human breast cancer cell line T47D, we demonstrate that prolactin (PRL), epidermal growth factor (EGF), or transforming growth factor-alpha (TGF-alpha) act on a mammary cell-specific enhancer at the extreme 5' end of the MMTV-LTR involving sequences -1094 through -858. PRL and either EGF or TGF-alpha exert concerted roles in this activation of these sequences. In contrast, using a plasmid construct lacking this mammary cell-specific enhancer, EGF or TGF-alpha, but not PRL, act synergistically with progesterone to induce CAT activity, indicating that the action of PRL on regulatory elements of the MMTV-LTR is restricted to this mammary cell-specific enhancer involving sequences -1094 through -858. A mobility shift assay was used to demonstrate that PRL, EGF or TGF alpha induce nuclear factors (MP4, MAF, and MGF) which bind directly to this mammary cell-specific enhancer element. PMID- 9202399 TI - Involvement of nitric oxide in the interferon-gamma-induced inhibition of growth hormone and prolactin secretion in anterior pituitary cell cultures. AB - In previous work it was shown that the immune cytokine interferon-gamma (IFN gamma) inhibits hormone secretion in anterior pituitary (AP) cell cultures, an action most likely mediated by folliculostellate (FS) cells. In the present study, we wanted to investigate whether nitric oxide (NO) is involved in this inhibitory action of IFN-gamma. NO synthase (NOS) inhibitors with affinity for the inducible (iNOS) and the constitutive (cNOS) isoform such as N(G)-monomethyl L-arginine (L-NMMA) and S-methyl-L-thiocitrulline (SMLT) dose-dependently blocked the inhibitory action of IFN-gamma on GHRH-stimulated GH secretion, and partially reversed the inhibitory effect on basal prolactin (PRL) release. In the absence of IFN-gamma these inhibitors significantly augmented basal PRL release and slightly enhanced GHRH-stimulated GH release. L-N6-(1-iminoethyl)lysine (L-NIL), a NOS inhibitor with preferential affinity for iNOS, abrogated the IFN-gamma effect on GHRH-stimulated GH secretion and partially reversed IFN-gamma inhibition of PRL release. However, L-NIL did not exert a stimulatory effect on basal PRL and GHRH-stimulated GH release by its own. 2,4-diamino-6 hydroxypyrimidine (DAHP), a NOS inhibitor by interfering with tetrahydrobiopterin (BH4) cofactor availability, showed the same activity profile as L-NIL. NOS inhibitors blocked or reduced the production of NO as detected by measuring nitrite (NO2-) levels in AP cell cultures and cGMP levels in the NO-reporter cell line RFL-6. The NOS inhibiting action of L-NMMA was confirmed by competition experiments with the natural NOS substrate L-arginine. Thus, in culture medium with lower amounts of L-arginine, L-NMMA blocked the IFN-gamma-induced inhibition of GHRH-stimulated GH release at a lower dose. The inhibition of PRL and GH release by IFN-gamma was markedly reduced in L-arginine-depleted medium. The NO donor sodium nitroprusside (SNP) mimicked the inhibitory action of IFN-gamma on GHRH-stimulated GH and basal PRL release. Similarly to IFN-gamma, SNP did not affect basal GH release. As previously reported, inhibition by IFN-gamma occurred only in AP cell populations containing a minimal proportion of FS cells. As studied in different cell populations obtained by unit gravity sedimentation in a serum albumin gradient, L-NMMA reversed the IFN-gamma effect in the same populations enriched in FS cells. Interestingly, in the absence of IFN-gamma L NMMA strongly stimulated basal PRL release in the population most enriched in FS cells. It is concluded that IFN-gamma through activation of the iNOS pathway probably in FS cells enhances the production of NO and that this effect is responsible for the inhibitory action of IFN-gamma on GHRH-stimulated GH release and partially for the IFN-gamma-induced decrease in basal PRL release. On the other hand, NO, likely produced by cNOS, appears to exert a tonic inhibitory effect on GHRH-stimulated GH and basal PRL release. It seems therefore that low amounts of NO produced constitutively may take charge of subtle physiological adaptations, and higher levels of NO produced by iNOS under the influence of IFN gamma may attenuate PRL and GH release during emergency conditions of immune and inflammatory reactions. PMID- 9202400 TI - CYP2C7 expression in rat liver and hepatocytes: regulation by retinoids. AB - Rats deficient in vitamin A express low levels of P4502C7 mRNA in the liver. Administration of all-trans retinoic acid (at-RA) or growth hormone (GH) to deficient animals only partially restored the expression whereas the combined treatment returned the P4502C7 mRNA levels to that observed in normal rats. That a retinoid is the predominant inducer of P4502C7 at the cellular level is evident from studies performed with primary hepatocytes, but it became clear that GH is a prerequisite for the vitamin A effect in vivo. The at-RA induction of P4502C7 mRNA in primary rat hepatocytes was inhibited by ketoconazole, an inhibitor of P450 activity, and by cycloheximide, blocking ongoing protein synthesis. In contrast, the at-RA induction of RAR-beta2 mRNA was not affected by any of these compounds. This could indicate previously not recognized mechanisms of at-RA action. Interestingly, at-4-oxo-RA, an at-RA metabolite formed by a P450 catalyzed reaction, also induced P4502C7 mRNA. Induction of P4502C7 mRNA by the retinoic acid receptor (RAR) selective agonist TTNPB indicated that this pathway is preferred over the retinoid X receptor (RXR) pathway. In addition, analysis of RA metabolites in liver cell extracts revealed the formation of several as yet unidentified metabolites. The formation of some of these metabolites was inhibited by ketoconazole and they could therefore constitute potential inducers of CYP2C7. We suggest that metabolism of at-RA, possibly by a P450 enzyme, is an important step in the at-RA induction of P4502C7. PMID- 9202401 TI - Identification of type 1 5alpha-reductase in myelin membranes of male and female rat brain. AB - The formation of the 5alpha-reduced metabolites of testosterone (T) and of progesterone (P) is a very active process in the brain, since the enzyme 5alpha reductase (5alpha-R) is present in almost any central nervous system (CNS) structure. A particularly elevated 5alpha-R activity has been shown in myelin sheaths. Two isoforms of the enzyme have been cloned, with different localisation as well as different biochemical properties. The present study was performed to determine whether both isoforms of the 5alpha-R, or only one of them, are/is responsible for the enzymatic activity observed in myelin. Kinetic analyses have been performed on purified myelin membranes prepared from the male or female rat brain, using both T and P as substrates. The 5alpha-R present appears to possess a pH optimum at basic values. The Vmax values obtained in the Lineweaver-Burk analysis were comparable in male and female preparations independently on whether T or P were used as the substrates, suggesting that a single enzymatic form is present in all samples examined; the Km obtained using [14C]T (Km: male 1.14 microM; female 1.46 microM) or [14C]P (Km: male 0.5 microM; female 0.64 microM) as substrates, were in good agreement with those obtained for the recombinant type 1 isoform. These data suggest that the type 1 isoform is the most relevant 5alpha-R present in myelin. To confirm this, a new polyclonal antibody was raised against the type 1 5alpha-R enzymatic protein, and used in immunohistochemical studies. The experiments were performed on the optic nerve, a myelinated structure very rich in 5alpha-R activity and the results clearly indicated the presence of a specific type 1 enzyme immunoreactivity in the myelin sheaths of axons. PMID- 9202402 TI - Inflammatory cytokines and type I 5'-deiodinase expression in phi1 rat liver cells. AB - Administration of tumour necrosis factor-alpha (TNF alpha), interleukin-1beta (IL 1beta) and interleukin-6 (IL-6) to animals and humans results in changes in circulating thyroid hormone concentrations similar to those seen in non-thyroidal illness (NTI). Inflammatory cytokines have been postulated as mediators of the euthyroid sick syndrome by inhibiting type 1 5'-deiodinase (5'D-I) enzyme activity. We have investigated direct effects of cytokines upon 5'D-I expression, measuring changes in 5'D-I enzyme activity and mRNA in phi1 rat liver cells. All three cytokines stimulated 5'D-I enzyme activity: TNF alpha 326 +/- 43% (100% in controls, mean + S.E.M., n = 9, P < 0.01 by ANOVA), IL-1beta 297 +/- 8% and IL-6 272 +/- 25%. Co-incubation with cycloheximide abolished stimulation by each cytokine. Kinetic analysis revealed that stimulation of 5'D-I enzyme activity was a result of significantly increased Vmax, (P < 0.01 by ANOVA) with Km relatively unchanged. 5'D-I mRNA abundance was not significantly changed following treatment by any of the three cytokines. These findings do not support the hypothesis that inflammatory cytokines may mediate the euthyroid sick syndrome by causing inhibition of 5'D-I activity. PMID- 9202403 TI - The last proline of Box 1 is essential for association with JAK2 and functional activation of the prolactin receptor. AB - The interaction of prolactin (PRL) with its receptor leads to activation of the tyrosine kinase, Janus kinase 2 (JAK2). In the cytoplasmic juxtamembrane region, a short segment (Box 1) which is conserved in other receptors of the PRL/growth hormone (GH)/cytokine receptor family, is required for signal transduction. To assess the contribution of the different amino acids of Box 1, individual alanine substitutions of all residues, grouped substitution of four prolines (4PA mutant) and individual leucine replacement of the two last prolines (P248L and P250L mutants) were introduced. Here we show that P250L and 4PA (i) inhibit PRL-induced transactivation of a luciferase reporter governed by a beta-caseine gene promoter; (ii) decrease in JAK2 tyrosine kinase activity in biotinylated-PRL precipitates; (iii) impair the interaction between PRLR and JAK2, as evidenced by lack of co-immunoprecipitation, (iv) and prevent the activation of signal transducer and activator of transcription (Stat) as determined by absence of tyrosine phosphorylation of Stat5. Our data suggest that the Box 1 region of the PRL receptor and particularly the last proline is critical for JAK2 association and subsequent activation. These results support the notion that the tyrosine kinase JAK2 is implicated in activation of downstream protein effectors such as Stat5, which are involved in transcription of PRL-responsive genes. PMID- 9202404 TI - Stimulation of the angiotensin II type I receptor on bovine adrenal glomerulosa cells activates a temperature-sensitive internalization-recycling pathway. AB - Angiotensin II (Ang II) is an important regulator of aldosterone production by bovine adrenal glomerulosa cells. On these cells Ang II interacts with the AT1 receptor that is coupled to a G protein controlling the activity of phospholipase C. A primary culture of bovine adrenal glomerulosa cells was used to study the internalization-recycling mechanism of the AT1 receptor after stimulation with Ang II. When cells were pretreated with 10 nM Ang II for 30 min at 37 degrees C and binding studies were performed at 12 degrees C we observed a 48% loss in [125I]Ang II binding. Scatchard analysis revealed that this loss in binding translated into a decreased affinity of the AT1 receptor without any loss in the total amount of binding sites. Under the same conditions an important internalization of [125I]Ang II was invariably observed. These observations suggest that a mechanism was at work to recycle the internalized receptors to the cell surface during the binding studies. Following internalization we indeed observed an externalization of [125I]Ang II. This phenomenon relatively rapid at 37 degrees C was much slower at 12 degrees C and completely inhibited at 4 degrees C. When cells were pretreated with 10 nM Ang II for 30 min at 37 degrees C binding assays at 4 degrees C no longer revealed a loss of binding affinity but rather a 54% reduction in the total amount of binding sites. The maximal binding capacity could be recovered during incubations at 12 degrees C. These results reveal the existence of a dynamic recycling process for the AT1 receptor. In accordance with this interpretation the phenomenon was blocked by monensin, a known inhibitor of receptor recycling. These studies suggest that the stimulation of the AT1 receptor sets in motion an internalization-recycling process that seems to be a fundamental aspect of the AT1 receptor transduction mechanism. PMID- 9202406 TI - MCF-7 and T47D human breast cancer cells contain a functional peroxisomal response. AB - Peroxisome proliferator-activated receptors (PPARs) are ligand-activated nuclear receptors that regulate transcription of target genes. Since attempts have been made to correlate the ingestion of high-fat diets, itself a peroxisome proliferator, with the occurrence of breast cancer, we set about to determine if human breast cancer cells contained a functional PPAR. In this report we demonstrate the presence of an mRNA in two breast cancer cell lines (MCF-7 and T47D) which is specifically recognized by a mouse PPARgamma2 probe. Furthermore, in gel shift assays a consensus PPAR response element (PPRE) was specifically bound by nuclear extracts from MCF-7 cells and was further retarded by antibodies raised to mouse PPARgamma. Finally, when transfected with a PPRE-luciferase transcriptional reporter construct, transcription was increased in response to activators of PPAR and its dimmeric partner the retinoic acid X receptor (RXR). These data indicate that peroxisomal proliferators are capable of mediating transcription in human breast cells and suggest the possibility of a physiological role in the breast. PMID- 9202405 TI - Characterization of an intronic hormone response element of the rat liver/skeletal muscle 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene. AB - The glucocorticoid response element of the rat liver/skeletal muscle 6- phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene was characterized. The element is composed of two tandem hormone receptor binding sites separated by 12 base pairs. Addition of dexamethasone to HeLa cells transiently transfected with a chloramphenicol acetyl transferase (CAT) reporter plasmid containing the hormone response element and cotransfected with glucocorticoid receptor stimulated transcription 24-fold in an orientation- and position-independent manner. Deletion or mutation of essential G/C pairs of the distal binding site abolished hormone-stimulated CAT activity, whereas deletion or mutation of the proximal binding site decreased the hormone-stimulated response only slightly. Mutation of both distal and proximal binding sites resulted in complete loss of hormone-stimulated CAT activity. Experiments carried out using testosterone and progesterone with their respective receptors revealed qualitatively similar results to those seen with glucocorticoid. Binding of glucocorticoid receptor or androgen receptor DNA binding domains to the hormone response element, visualized by gel mobility shift, was unaffected in the proximal binding site mutant, markedly decreased in the distal binding site mutant, and abolished in the double mutant. In gel mobility shift analysis of separate distal and proximal binding sites, only the native distal site demonstrated high affinity binding to glucocorticoid and androgen receptor DNA binding domains. The results demonstrate that this element is responsible for glucocorticoid, androgen, and progesterone stimulation of transcription of the 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase gene and that the distal receptor binding site is dominant. PMID- 9202407 TI - A synthetic approach to the c-series gangliosides containing sialyl-alpha(2- >8)sialyl-alpha(2-->8)sialic acid: synthesis of ganglioside GT4, alpha(2-->6) GT4 and GT3. AB - Trimeric sialic acid [Neu5Ac alpha(2-->8)Neu5Ac alpha(2-->8)Neu5Ac, 1] residue containing gangliosides, GT4, alpha(2-->6)GT4 and GT3, have been synthesized for the first time. Methyl [phenyl] 5-acetamido-8-O-[5-acetamido-8-O-(5-acetamido-4, 7, 8, 9-tetra-O-acetyl-3, 5-dideoxy-D-glycero-alpha-D-galacto-2 nonulopyranosylono-1", 9'-lactone)-4,7-di-O-acetyl-3,5-dideoxy-D- glycero-alpha-D galacto-2-nonulopyranosylono-1',9-lactone]-4,7-di- O-acetyl -3,5-dideoxy-2-thio-D glycero-D-galacto-2-nonulopyranosid]onate (3) was prepared from 1, via lactonization, methyl esterification of the carboxyl group at the reducting end, O-acetylation and conversion of the anomeric acetoxy group into a phenylthio group. Iodonium-promoted glycosylation of 3 with 2-(trimethylsilyl)ethyl 2,6-di-O benzyl-beta-D-galactopyranoside (5), 2-(trimethylsilyl)ethyl 3-O-benzyl-beta-D galactopyranoside (6), 2-(trimethylsilyl)ethyl 2-O-benzoyl-3-O-benzyl-beta-D galactopyranoside (9), and 2-(trimethylsilyl)ethyl 2, 3-di-O-benzyl-beta-D galactopyranoside (11) gave the corresponding tetrasaccharides (13-15, 17) having the (Neu5Ac)3-Gal structure. The peracylated oligosaccharides 18 and 24 derived from 13 and 17, and the previously reported lactose derivative 29 were converted into the alpha-trichloroacetimidates 20, 26 and 31, and coupled with (2S,3R,4E)-2 azido-3-O-benzoyl-4-octadecene-1,3-diol (21) to afford the corresponding beta glycosides 22, 27 and 32. These protected azidosphingosine derivatives were each transformed into the target gangliosides GT4, alpha(2-->6)GT4 and GT3 via selective reduction of the azido group, subsequent coupling with octadecanoic acid, O-deacylation and saponification of the methyl ester and lactone groups. PMID- 9202408 TI - Rhamnogalacturonan II from the leaves of Panax ginseng C.A. Meyer as a macrophage Fc receptor expression-enhancing polysaccharide. AB - A complex pectic polysaccharide (GL-4IIb2) has been isolated from the leaves of Panax ginseng C.A. Meyer, and shown to be a macrophage Fc receptor expression enhancing polysaccharide. The primary structure of GL-4IIb2 was elucidated by composition. 1H NMR, methylation, and oligosaccharide analyses. GL-4IIb2 consisted of 15 different monosaccharides which included rarely observed sugars, such as 2-O-methylfucose, 2-O-methylxylose, apiose, 3-C-carboxy-5-deoxy-L-xylose (aceric acid, AceA), 3-deoxy-D-manno-2-octulosonic acid (Kdo), and 3-deoxy-D-lyxo 2-heptulosaric acid (Dha). Methylation analysis indicated that GL-4IIb2 comprised 34 different glycosyl linkages, such as 3,4-linked Fuc, 3- and 2,3,4-linked Rha, and 2-linked GlcA, which are characteristic of rhamnogalacturonan II (RG-II). Sequential degradation using partial acid hydrolysis indicated that GL-4IIb2 contained alpha-Rhap-(1-->5)-Kdo and Araf-(1-->5) Dha structural elements, an AceA-containing oligosaccharide, and uronic acid-rich oligosaccharide chains in addition to an alpha-(1 -->4)-galacturono-oligosaccharide chain. FABMS and methylation analyses suggested that the AceA-containing oligosaccharide was a nonasaccharide in which terminal Rha was additionally attached to position 3 of 2 linked Arap of the octasaccharide chain observed in sycamore RG-II. Component sugar and methylation analyses assumed that the uronic acid-rich oligosaccharides possessed a similar structural feature as those in sycamore RG-II. GL-4IIb2 had a larger molecular mass (11,000) than sycamore RG-II (approximately 5000). PMID- 9202409 TI - Lectin-deficient ricin toxin intoxicates cells bearing the D-mannose receptor. AB - Ricin toxin with genetic or chemical modification of lectin sites has been previously reported to show markedly reduced cytotoxicity to cells following uptake by several receptors including the mannose receptor. Investigators have hypothesized that an intracellular galactoside-binding function was required for optimal intracellular targeting of ricin for these receptors. We have prepared insect-derived mutant ricin toxin B chain (RTB) with modifications of three lectin side domains (1 alpha, 1 beta, and 2 gamma) yielding a 1000-fold reduced galactoside avidity. After reassociation with plant RTA, the recombinant heterodimer and plant ricin were tested for cytotoxicity on mammalian cells expressing (mouse peritoneal macrophages, J774E cells, and MMR61 cells) or not expressing (KB cells) the D-mannose receptor. Receptor expression was confirmed by immunofluorescence microscopy. Lactose was included in the media to block cell surface galactoside binding, and mannan was added as a control in each experiment to confirm mannose receptor-specific targeting. Plant ricin A chain (RTA) and E. coli-derived RTA were also tested for cytotoxicity on J774E and KB cells. Both wild-type and lectin-deficient ricin displayed mannose-receptor mediated cell cytotoxicity. This is the first report of a genetically modified ricin showing that RTB intracellular galactose binding activity is not required for ricin cytotoxicity. Sensitivity of mannose-receptor bearing cells, but not control cells, to mannosylated RTA, but not unglycosylated RTA, confirmed these observations. These results imply fusion toxins employing ricin can be prepared with maximal reductions in normal tissue binding. PMID- 9202410 TI - Preparation and biological activity of manno- and galacto-validamines, new 5a carba-glycosylamines as alpha-glycosidase inhibitors. AB - Manno- and galacto-validamines, which are epimers of validamine, were semi synthesized by the configurational inversion of validamine, a pseudo-sugar analogue of alpha-D-gluco-pyranose that has inhibitory activity for alpha glucosidases. The inhibitory activities of these analogues were determined against several mannosidases and galactosidase. Manno-validamine shows potent inhibition for the alpha-mannosidases (competitive. K(i) = 4.6 x 10(-5) M for jack beans, and competitive, Ki = 2.8 x 10(-5) M for almonds), and galacto validamine shows weak inhibition for the alpha-galactosidases (coffee bean and E. coli). The inhibitory effect of the epimers on the N-linked oligosaccharide processing mannosidases involved in glycoprotein biosynthesis and lysosomal mannosidase from rat liver were also examined. Mannovalidamine shows potent inhibition on the endoplasmic reticulumal alpha-mannosidase (competitive, K(i) = 1.2 x 10(-6) M), Golgi mannosidases IA, II (competitive, K(i) = 2.8 x 10(-5) M), and lysosomal alpha-mannosidase (competitive, Ki = 1.7 x 10(-5) M). PMID- 9202411 TI - Distribution of highly repeated DNA sequences in species of the genus Secale. AB - The presence and distribution of the most important highly repetitive DNA sequences of rye in cultivated and wild species of the genus Secale were investigated using fluorescence in situ hybridization. Accurate identification of individual chromosomes in the most commonly recognized species or subspecies of the genus Secale (S. cereale, S. ancestrale, S. segetale, S. afghanicum, S. dighoricum, S. montanum, S. montanum ssp. kuprijanovii, S. africanum, S. anatolicum, S. vavilovii, and S. silvestre) was achieved using three highly repetitive rye DNA sequences (probes pSc119.2, pSc74, and pSc34) and the 5S ribosomal DNA sequence pTa794. It is difficult to superimpose trends in the complexity of repetitive DNA during the evolution of the genus on conclusions from other cytogenetic and morphological assays. However, there are two clear groups. The first comprises the self-pollinated annuals S. silvestre and S. vavilovii that have few repeated nucleotide sequences of the main families of 120 and 480 bp. The second group presents amplification and interstitialization of the repeated nucleotide sequences and includes the perennials S. montanum, S. anatolicum, S. africanum, and S. kuprijanovii, as well as the annual and open pollinated species S. cereale and its related weedy forms. The appearance of a new locus for 5S rRNA in S. cereale and S. ancestrale suggests that cultivated ryes evolved from this wild weedy species. PMID- 9202412 TI - Chromosomal localization of the human liver form cytochrome c oxidase subunit VIIa gene. AB - The chromosomal loci corresponding to human cytochrome c oxidase (COX) subunit VIIa Liver (VIIa-L) isoform genes were determined utilizing a combined approach of genomic cloning, in situ hybridization, and somatic hybrid genetics. In contrast to the proposal of E. Arnaudo et al. (Gene (Amst.), 119: 299-305, 1992) that COX VIIa-L sequences are located on chromosomes 4 and 14, we found that COX VIIa-L related sequences reside on chromosome 6, while an additional COX VIIa-L cross-reacting sequence psi-gene) was located on chromosome 4. PMID- 9202413 TI - Cloning, characterization, and chromosomal localization of human liver form cytochrome c oxidase subunit VIa related genes. AB - The chromosomal location of human cytochrome c oxidase (COX) subunit VIa Liver (VIa-L) isoform related sequences has been determined by a combination of in situ hybridization and analysis of human-hamster somatic cell hybrid panels. COX VIa-L related sequences were present on chromosomes 6 and 12. It has been verified that at least two COX VIa-L genes are on chromosome 6, one of which is a pseudogene. In total, four COX VIa-L related sequences have been cloned and their nucleotide sequences analyzed. At least three of the sequences represent pseudogenes; their relatedness to the COX VIa-L cDNA is discussed. PMID- 9202414 TI - The evolution of species-type specificity in the global DNA sequence organization of mitochondrial genomes. AB - Prokaryote genomes and nuclear genomes of eukaryotes have a global DNA sequence organization that is species type specific, determined primarily by nearest neighbor nucleotide associations, and independent of gene function and sequence length. The determinants of such a global structure have remained largely uncharacterized. The monophyletic and endosymbiotic origin of mitochondria permit examination of the influence of evolutionary time and host species type. Different global structures were seen among (i) protozoan and plant (ii) fungal, (iii) algal (iv) nematode, (v) echinoderm, (vi) insect, and (vii) vertebrate species followed examination of 28 complete mitochondrial genomes using chaos representation and measure of short-sequence representation. The mitochondrial genomes have biases in single-nucleotide and dinucleotide representation, specifically, an overrepresentation of A and T nucleotides and CC/GG and AG/CT dinucleotides and a deficiency of CG dinucleotides, in all but one genome. Dinucleotide representation is similar among (i) mitochondrial genomes of more closely related species; (ii) mitochondrial genomes and the Mycoplasma capricolum genome, a proposed progenitor of mitochondrial genomes; and (iii) mitochondrial genomes of diverse species, more so than between the mitochondrial and the nuclear genome of the same or a closely related species. It is hypothesized that sufficient evolutionary time has permitted host-specific constraints to affect nuclear and mitochondrial genomes and that different species type specific constraints influence nuclear and mitochondrial genome global structure. PMID- 9202415 TI - Comparative evaluation of within-cultivar variation of rice (Oryza sativa L.) using microsatellite and RFLP markers. AB - The objective of this study was to determine an efficient way of detecting within cultivar variation in rice varieties obtained from national and international germplasm collections. Seventy-one rice cultivars were evaluated for within cultivar variation using a combination of phenotypic, RFLP, and microsatellite or simple sequence length polymorphism (SSLP). Variation between individuals within and accession and between duplicate accessions within a cultivar was detected even in cultivars that had been purified by phenotypic evaluation. Landrace cultivars were more heterogeneous and displayed a larger number of both RFLP and SSLP alleles than did modern cultivars. Microsatellite markers detected a greater number of alleles and were able to discriminate between even closely related individuals more efficiently than RFLPs. Some microsatellite markers were more informative than others for assessing genetic diversity. Single markers revealed 5.6-61.1% of the total variation detected by the 10 SSLP markers. Some marker combinations were complementary, providing more information than others. Several combinations of 4 SSLP markers detected as much as 94% of the total within cultivar variation detected by the 10 SSLP markers. These results suggest that the use of four well-chosen microsatellites would be an efficient method for evaluating the heterogeneity of rice accessions. PMID- 9202416 TI - Aneuploid marker assignment in hexaploid oat with the C genome as a reference for determining remnant homoeology. AB - Nullisomic lines of hexaploid oat Avena sativa L. (2n = 6x - 2 = 40, AACCDD) cultivar Sun II were used to assign 134 DNA sequences to 10 chromosome-associated syntenic groups. A limited set of ditelosomic lines allowed localization of subsets of these sequences to six chromosome arms. Advantages of using such aneuploids in mapping are in the assignment of gene families, monomorphic RFLP sequences, and oat linkage groups to chromosomes. The published hexaploid oat RFLP linkage map has 38 linkage groups, 17 more than expected on the basis of the haploid chromosome number. Using nullisomics, eight linkage groups were assigned to five physical chromosomes; using ditelosomics, three of these linkage groups were assigned to their respective chromosome arms. The A- and D-genome chromosome sets of oat are indistinguishable from each other based on different staining and genomic in situ hybridization techniques, while C-genome chromosomes are distinct. Because chromosomal rearrangements such as translocations and inversions have played an important role in the evolution of hexaploid oat, the distinction of C-genome chromosomes can be used to determine remnant homoeologous segments that exist in the other two genomes. Among the 10 syntenic groups identified, six chromosomes showed sequence homoeology believed to represent segmental homoeologous regions. Owing to various evolutionary forces, segmental homoeology instead of whole chromosome homoeology appears to best describe the genome organization in hexaploid oat. PMID- 9202417 TI - Sex and meiosis in autotetraploid Pacific oyster, Crassostrea gigas (Thunberg). AB - Sex and meiosis were studied in induced autotetraploids of the Pacific oyster (Crassostrea gigas Thunberg) and were compared with sex and meiosis in autotriploids and normal diploids. Tetraploid oysters reached sexual maturity at 1 year of age in an approximately 1:1 sex ration. In contrast with the abnormally high frequency of hermaphrodites among triploids, tetraploids had about the same level of hermaphrodites as normal diploids. Fecundity of tetraploids was comparable to that of normal diploids, differing from the greatly reduced fecundity of triploids. Homologous chromosomes synapsed predominantly as trivalents in eggs from triploids and as quadrivalents in eggs from tetraploids. After fertilization, eggs from tetraploids and triploids went through two meiotic divisions, as normal eggs did. The average gamete chromosome number was 10.0 for diploids and 19.9 for tetraploids. The distribution of gamete chromosome numbers from triploids suggested that the extra chromosome in the trivalent segregated randomly during anaphase I. In tetraploids, however, the two extra chromosomes in the quadrivalents did not segregate independently and, instead, they preferentially cosegregated to opposite poles producing balanced gametes. These results suggest that mechanisms may exist to weight, balance, and equally distribute quadrivalents, possibly through mitotic force and tension. Errors in chromosome balancing in normal meiosis may result in nondisjunction, which is the primary cause of human aneuploidy. PMID- 9202418 TI - The apoptotic cysteine protease CPP32. AB - CPP32 (also called Yama and apopain) is a member of a growing family of cysteine proteases which includes the interleukin-1 beta-converting enzyme (ICE) and the product of the Caenorhabditis elegans cell death gene ced-3. CPP32 has been consistently implicated as a key protease of the ICE/CED-3 family that is activated in response to a variety of death stimuli. Active CPP32 consists of P17 and p12 subunits derived from a 32 kDa pro-enzyme. This activation can be mediated by some ICE-like proteases and the cytotoxic T-cell (CTL) protease granzyme B. Once activated, CPP32 can process some of the other ICE family members and cleaves several cellular proteins in apoptotic cells. Inhibitors of CPP32 and other ICE-like proteases are potent inhibitors of apoptosis and promise to be important therapeutic molecules for the treatment of diseases, such as neurodegenerative and autoimmune disorders, that arise from excessive ell death. PMID- 9202419 TI - Lyn, a src-like tyrosine kinase. AB - Lyn is a member of the src family of non-receptor protein tyrosine kinases that is predominantly expressed in haematopoietic tissues. Like all members of the src family, lyn is thought to participate in signal transduction from cell surface receptors that lack intrinsic tyrosine kinase activity. It is associated with a number of cell surface receptors including the B cell antigen receptor and Fc epsilon RI. Lyn deficient mice develop autoimmune disease characterised by autoantibodies in serum and the deposition of immune complexes in the kidney, a pathology reminiscent of systemic lupus erythematosus. Lyn deficient mice also have impaired signalling involving Fc epsilon RI in mast cells, resulting in defective allergic responses. PMID- 9202420 TI - Neuronal laminins and their cellular receptors. AB - The laminins are a family of extracellular matrix glycoproteins expressed throughout developing neural tissues. The laminins are potent stimulators of neurite outgrowth in vitro for a variety of cell types, presumably reflecting an in vivo role in stimulating axon outgrowth. In recent years, the laminins have been shown to occur in several distinct isoforms; currently, the precise functional differences between the laminin variants are not well understood. A variety of neuronal surface receptors have been identified for one laminin isoform, laminin-1. These receptors include several members of the integrin family, as well as non-integrin laminin-binding proteins such as LBP-110, the 67 kDa laminin-receptor, alpha-dystroglycan, and beta 1,4 galactosyltransferase. Little is currently known about receptors for other laminin isoforms. PMID- 9202421 TI - Growth factor-dependent phosphoinositide signalling. AB - A wide variety of messages, in the form of diffusible growth factors, hormones and cytokines, are carried throughout multicellular organisms to coordinate important physiological properties of target cells, such as proliferation, differentiation, migration, apoptosis and metabolism. Most messengers bind to cognate receptors on target cells, which initiate a characteristic cascade of reactions within the cell, ultimately leading to the desired response. The cellular response is defined by the combination of signalling components whose individual activity depends upon the number and type of surface receptors. Consequently the responses of different cell types to one or more stimuli can be quite disparate. A molecular understanding of the signalling pathways employed by each type of receptor therefore underlies the ability to rationalize many cellular functions and to correct disfunctions. As a well studied example of the primary signalling events that take place on the cytoplasmic leaflet of the plasma membrane following receptor activation, we will discuss how the widely expressed receptor for epidermal growth factor (EGF) causes the phosphorylation and hydrolysis of a signalling precursor, the membrane lipid phosphatidylinositol. This paradigm will be used to illustrate certain general principles of signalling, including formation of multienzyme complexes, compartmentation of second messengers and intermediates, and cross-talk between different signalling pathways. PMID- 9202422 TI - Mechanisms of active oxygen species reduction by non-steroidal anti-inflammatory drugs. AB - Many forms of active oxygen have been suggested to participate in the course of inflammation. Anti-inflammatory drugs have been considered to function as active oxygen inhibitors. However, detailed mechanisms for such inhibitory activity remain unclear because of little well established methods to study inhibitory effect of anti-inflammatory drugs on active oxygen species. In this report, the author investigated four non-steroidal anti-inflammatory drugs, including acetaminophen, sodium salicylate, naproxen and flurbiprofen, their elimination and inhibition ability of active oxygen, using of the electron spin resonance spin-trapping method and the horseradish peroxidase method. In this experiment as active oxygen models, superoxide was evolved from a hypoxanthine-xanthine oxidase reaction system, and hydrogen peroxide by the spontaneous dismutation reaction. The data here show that the amount of superoxide was reduced in the manner of concentration of non-steroidal anti-inflammatory drugs in the reaction. Kinetic studies for these reaction showed that acetaminophen and sodium salicylate reacted with superoxide competitively, whereas naproxen and flurbiprofen did not. Analysis of generation of hydrogen peroxide formed by the spontaneous dismutation of superoxide derived from the reaction system revealed that hydrogen peroxide was increased by acetaminophen and decreased by sodium salicylate, naproxen and flurbiprofen. PMID- 9202423 TI - Extracellular calcium stimulates Na(+)-dependent putrescine uptake in B16 melanoma cells. AB - The regulation of putrescine transport in difluoromethylornithine-treated B16 melanoma cells by extracellular Ca2+ has been investigated. It was found that physiological concentrations of Ca2+ were essential for optimum uptake of putrescine and spermidine, Mg2+, albeit at higher concentrations, also could potentiate polyamine transport. The maximum rate of putrescine uptake increased from 1698 +/- 67 pmol/min/min/mg DNA in the absence of Ca2+ to 3100 +/0 98 pmol/min/mg DNA in the presence of 0.5 mM Ca2+. There was no change in Km. While Ca2+ enhanced transport of both putrescine and spermidine it did not affect the uptake of deoxyglucose, thymidine or leucine. Putrescine did not alter Ca2+ fluxes suggesting that the two cations do not share a common transport system. The effects of Ca2+ on putrescine uptake appeared to be mediated extracellularly firstly because Ca2+ did not potentiate putrescine uptake in the presence of A23187 and secondly, because the effects of Ca2+ were completely inhibited by the lanthanide Tb3+, which binds to calcium-dependent proteins and does not readily cross biological membranes, Ca2+ did not affect putrescine transport in the absence of extracellular Na+. Moreover, the rate of putrescine uptake in the absence of Ca2+ was similar to that in the absence of extracellular Na+. The results from this study indicate that polyamine transport is stimulated by extracellular Ca2+ and suggest that Ca2+ is required for activity of the Na(+) dependent transporter only. This transporter appears to possess a regulatory binding site for divalent cations. PMID- 9202424 TI - Presence of Vicia graminea lectin- and Vicia unijuga lectin-binding (Vgu) glycoprotein in human amniotic fluid and some of its chemical and serological properties. AB - Vicia graminea- and Vicia unijuga-binding glycoprotein (Vgu glycoprotein) has been reported as a malignant tumor-associated antigen, which is found in various kinds of malignant tumor-tissues and ascitic and cyst fluids of malignant tumor patients, but not found in 20 kinds of normal human tissues. The glycoprotein reacts with anti-N lectins of Vicia-graminea (VGA) and Vicia unijuga (VUA), but does not react with anti-blood group M and N sera. The existence of Vgu glycoprotein in human amniotic fluid is reported here. A perchloric acid-soluble glycoprotein fraction (PASF) was prepared from human amniotic fluid (AF-PASF). Sialoglycoprotein was then separated from AF-PASF by consecutive gel filtration on Sephacryl S-300, HPLC on Asahipak GS-710 and affinity chromatography in VUA conjugated formyl-cellulofine columns. The sialoglycoprotein with 9.6% (w/w) carbohydrate content showed a molecular mass of 2940 kDa estimated by HPLC gel filtration. The sialoglycoprotein showed reactivity with VGA and VUA, but did not react with anti-M and -N sera. In addition, the sialoglycoprotein reacted with lectins of Ulex europeus, Arachis hypogaea, Maclura pomifera, Canavalia ensiformis, Ricinus communis, Phaseolus vulgaris, Sambucus nigra and Sophora japonica. These results show that Vgu glycoprotein exists in human amniotic fluid, and that Vgu glycoprotein obtained from human amniotic fluid has a similar serological property to Vgu glycoproteins found in malignant tumor-tissues, but its chemical property is significantly different from that of malignant-tumor associated Vgu glycoproteins. PMID- 9202425 TI - The anti-Fc gamma RIII mAb 3G8 induces neutrophil activation via a cooperative actin of Fc gamma RIIIb and Fc gamma RIIa. AB - Human neutrophils express two types of Fc gamma receptors, the transmembrane Fc gamma RIIa and the glycan-phosphatidylinositol-anchored Fc gamma RIIIb, that show synergism in provoking a cellular response. To analyse further the requirements for this synergism to occur we used the monoclonal antibody 3G8, directed against Fc gamma RIII. This antibody is able to induce neutrophil activation, as measured by an increase in the intracellular free Ca2+ concentration and homotypic neutrophil aggregation, but only when the Fc part of the antibody is able to interact with Fc gamma RIIa. We observed that binding of the Fab parts of 3G8 mAb to two Fc gamma RIIIb molecules and binding of the Fc part to one Fc gamma RIIa molecule is required, because a bispecific antibody, 2B1, in which only one 3G8 Fab is present, did not induce neutrophil activation. Moreover, engagement of one Fc gamma RIIa molecule and two Fc gamma RIIb molecules on the same cell is instrumental to achieve activation of the mAb 3G8. The activation of neutrophils by the 3G8 antibody represents a further example of synergistic activation of neutrophils via Fc gamma receptors. PMID- 9202426 TI - Arylsulfatase A from human placenta possesses only high mannose-type glycans. AB - It has been shown that the concentration of arylsulfatase A increases in the body fluids of patients with some forms of cancer and the carbohydrate component of arylsulfatase A synthesized in tumor tissues and transformed cells undergoes increased sialylation, phosphorylation and sulfation. The specificity of changes in the glycosylation of glycoproteins in cancer is still unknown. To understand the significance of any changes in glycosylation of arylsulfatase A in cancer, it is important to know the structure of its carbohydrate component in normal tissue. Here, carbohydrate moieties of human placental arylsulfatase A were studied by sequential lectin affinity chromatography after enzymatic cleavage and labelling with tritiated sodium borohydride. Labelled oligosaccharides were subjected to ion exchange chromatography. The uncharged fraction and the neuraminidase treated charged fraction were further analysed using the lectins: Concanavalin A (Con A), Ricinus communis (RCA I), Triticum vulgaris (L-PHA) and Aleuria aurantia (AAL). The results indicated that 97% of the arylsulfatase A oligosaccharides were low molecular weight high mannose type glycans possessing up to 5 mannose residues. This was supported by the approximately 2.4 kDa decrease in the molecular weight of arylsulfatase. A subunits upon complete peptide N-glycosidase F deglycosylation, as shown using SDS-PAGE. The remaining 3% of the arylsulfatase A oligosaccharides were of the high mannose type, possessing more than 5 mannose residues. Most (97.5%) of the glycans were uncharged, while 2.5% were charged. Neuraminidase treatment of the latter did not remove the charge, suggesting the presence of phosphate or sulfate residues. This study, of arylsulfatase A oligosaccharides separated from the protein part, shows that all glycans of the enzyme from human placenta are of the high mannose type. PMID- 9202427 TI - Recombinant rat liver S-adenosyl-L-methionine synthetase tetramers and dimers are in equilibrium. AB - Rat liver S-adenosyl-L-methionine synthetase is present in two oligomeric forms, tetramers and dimers, with different substrate kinetics and regulation. In vivo the relative amounts of both forms may change in some instances. The basis of this regulatory mechanism is not known. When rat liver cDNA was used to express the protein in Escherichia coli the two oligomeric forms were found. Gel filtration chromatography of the purified recombinant enzyme suggested that these two isoforms might be in equilibrium. This was confirmed by kinetic experiments which showed that the specific activity of the enzyme was dependent on the protein concentration. From these experiments, apparent equilibrium constants of (5.6 +/- 0.4) x 10(5) M-1 and (3.5 +/- 0.9) x 10(5) M-1 were obtained at 2mM and 60 microM methionine concentrations, respectively. Using hydrophobic chromatography on phenyl-Sepharose to separate the tetrameric and dimeric forms, an equilibrium constant of (4.9 +/- 0.7) x 10(5) M-1 was calculated. A rate constant for the dissociation of the tetramer of k-1 = (8.1 +/- 0.4) x 10(-4) s-1 at 4 degrees C was also calculated using the same approach. In summary, we have shown that the rat liver S-adenosyl-L-methionine synthetase produced in bacterial cells is present in two oligomeric forms, tetramers and dimers, which are in equilibrium. This system might be useful for studying the dynamics and the regulation of the distribution of oligomeric forms in the mammalian liver. PMID- 9202428 TI - Folding and unfolding of delta-aminolevulinic acid dehydratase and porphobilinogen deaminase induced by uro- and protoporphyrin. AB - In all the cutaneous porphyrias, alterations in the heme pathway lead to an excessive production and accumulation of porphyrins. Absorption of light energy by circulating porphyrins induces reactive oxygen species generation, which provoke enzyme inactivation and protein structure changes. Protein structure alterations induced by porphyrins with different physico-chemical properties on delta-aminolevulinic acid dehydratase (ALA-D) and porphobilinogen deaminase (PBG D) were examined. The action of uroporphyrin (URO), a highly hydrophilic porphyrin, and protoporphyrin (PROTO), most hydrophobic, was tested. ALA-D and PBG-D were partially purified from bovine liver and exposed to URO or PROTO, both in the dark and under UV light. All experiments were performed in solution after removing the porphyrins. Treatment with 10 microM URO I or 10 microM PROTO IX reduced the activity of ALA-D and PBG-D. This effect increased with increasing time of exposure to porphyrins. Solubility of the enzymes in buffer containing 3 M KCl decreased with increasing time of porphyrin treatment; this may be because of exposure of hydrophobic residues that are normally shielded in the native protein structure. Tryptic digestion of ALA-D and PBG-D exposed to URO I or PROTO IX resulted in an increase of protein degradation products, indicating an enhanced susceptibility to proteolysis. Fluorescence emission of several enzymes aminoacids was greatly modified. The structural changes described were observed when the enzymes were exposed to porphyrins both in the dark or under UV light. However, they were more noticeable with UV light. These results suggest that porphyrins per se can act directly on protein structure and that this action may be enhanced by UV irradiation. PMID- 9202429 TI - RAS2/PKA pathway activity is involved in the nitrogen regulation of L-leucine uptake in Saccharomyces cerevisiae. AB - The aim of the present work is to study the participation of RAS2/PKA signal pathway in the nitrogen regulation of L-leucine transport in yeast cells. The study was performed on Saccharomyces cerevisiae isogenic strains with the normal RAS2 gene, the RAS2val19 mutant and the disrupted ras2::LEU2. These strains bring about different activities of the RAS2/PKA signal pathway, L-(14C)-Amino acid uptake measurements were determined in cells grown in a rich YPD medium with a mixed nitrogen source or in minimal media containing NH4+ or L-proline as the sole nitrogen source. We report herein that in all strains used, even in those grown in a minimal proline medium, the activity of the general amino acid permease (GAP1) was not detected. L-Leucine uptake in these strains is mediated by two kinetically characterized transport systems. Their KT values are of the same order as those of S1 and S2 L-leucine permeases. Mutation in the RAS2 gene alters initial velocities and Jmax values in both high and low affinity L-leucine transport systems. Activation of the RAS2/PKA signalling pathway by the RAS2val19 mutation, blocks the response to a poor nitrogen source whereas inactivation of RAS2 by gene disruption, results in an increase of the same response. PMID- 9202430 TI - The in vitro expression patterns of individual type I interferon genes in Newcastle disease virus infected murine splenocytes and fibroblasts. AB - Murine type I interferon levels present in mice sera following Newcastle disease virus infections are influenced by the If-1 locus. Sera interferon levels in C57BL/6 mice (If-1h allele) are 10- to 15-fold higher than in BALB/c mice (If 1(1) allele). The B6.C-H-28c strain, which carries BALB/c If-1(1) allele on C57BL/6 genomic background, has low interferon levels in sera. This study examined the expression of interferon alpha 1, alpha 4, alpha 5, alpha 6, alpha 9 and beta mRNAs at 7 hr after Newcastle disease virus infection of primary cells (splenocytes and mouse embryo fibroblasts) from C57BL/6, B6.C-H-28c and BALB/c mouse genotypes. Total RNA from these cells was reverse transcribed and all known type I interferon subtypes were amplified. The products were identified by differential hybridization to a panel of subtype specific oligonucleotides. The results show that the pattern of interferon subtypes examined in splenocytes did not differ between If-1h and If-1(1) allele carrying C57BL mice. However, when the genotype was different (BALB/c splenocytes) the pattern of type I interferon mRNAs seen was altered. This genotype-dependent expression was also seen in newcastle disease virus infected fibroblasts. Within a given mouse strain, there were also differences in the subtype response patterns detected in fibroblasts compared with those seen in splenocytes. In conclusion, the present study indicates that mouse genotype appears to be a major determinant of the subtype response pattern seen and tissue specific pattern differences are present within a given mouse genotype. PMID- 9202431 TI - Extracellular biotransformation potential in mouse airways. AB - The aims of this study were to determine if freshly isolated bronchiolar Clara cells retained biotransformation potential and whether components of phase 1 and/or phase 2 metabolism were present in the extracellular lining fluid of the lung airways. Approximately 550 microliters of lavage fluid was obtained from the mouse lung, which had been totally perfused of blood in order to facilitate the isolation of Clara cells from the same animal. Samples of acellular lavage fluid and aliquots of purified Clara cells were frozen at -20 degrees C prior to analysis. minimum numbers of cells and lavage fluid volumes, pooled from 2 CD-1 mice, were assayed for ethoxycoumarin-O-deethylase, NADPH-dependent cytochrome c reductase, glutathione-S-transferase(s) and non-protein sulphydryls (mostly reduced glutathione). Isolated Clara cells retained a high activity/level of all these parameters confirming their functional status for subsequent studies of xenobiotic metabolism in vitro. Acellular lavage fluid, from all the healthy animals, also contained mono-oxygenase, reductase and transferase activities and high non-protein sulphydryl levels suggesting that phase 1 and 2 reactions with xenobiotics could take place in the extracellular environment of the lung. Clara cells are known to undergo apocrine secretion and this removal of the apical cap containing secretory granules and smooth endoplasmic reticulum could account for the airway biotransformation potential that was observed. PMID- 9202432 TI - Ryanodine receptor binding constants in skeletal muscle, heart, brain and liver of the Mexican volcano mouse (Neotomodon alstoni alstoni; Rodentia:Cricetidae). Comparison with five other rodent species. AB - Equilibrium [3H]ryanodine binding assay was applied to total membrane fractions of six rodent species, including the Mexican volcano mouse Neotomodon alstoni alstoni, Wistar rat Rattus norvegicus albinus, golden hamster Mesocritus auratus, gerbil Meriones unguiculatus, guinea-pig Cavia porcellus, and ground squirrel Spermophillus mexicanus. The organs selected for this study were: skeletal muscle, heart, brain and liver. The constants derived from Scatchard analysis show slight variations in their Kd, ranging from 3 to 15 nM, except in the gerbil's skeletal muscle (38 nM) and the hamster's brain (27 nM). Remarkably, the Bmax calculated in guinea-pig muscle was as high as that reported for the rabbit fast twitch muscle (4.6 pmol/mg of protein) using the same membrane fraction preparation. For all the other skeletal muscles, Bmax was similar to the corresponding heart Bmax values (from 0.5 to 1 pmol/mg of protein). Gerbil cardiac Bmax was the highest (1.1 pmol/mg of protein). The ground squirrel was the rodent with more cerebral ryanodine binding sites (0.26 pmol/mg of protein), whereas the rat and the volcano mouse showed the lowest values (0.12 pmol/mg of protein). The richest sources of hepatic ryanodine receptor were the guinea-pig and rat livers (approximately equal to 0.35 pmol/mg of protein), whereas the lowest Bmax corresponded to the hamster liver (0.018 pmol/mg of protein). These results allow us to detect the similarities and differences of the ryanodine receptor binding constants from four different tissues of some of the rodents most widely used as biomedical laboratory animals. PMID- 9202433 TI - Non-enzymatic peroxidation of lipids isolated from rat liver microsomes, mitochondria and nuclei. AB - Studies were carried out to determine the level of lipoperoxidation ascorbate Fe2+ dependent on polar and neutral lipids isolated from rat liver microsomes, mitochondria and nuclei subjected to an incubation at 37 degrees C for 140 min. Three experimental approaches were used: recording of low-level chemiluminescence, determination of fatty acid composition and measurement of radioactivity of lipidic species of each kind of membrane during the peroxidation process. Membrane light emission decreased in the order microsomes > mitochondria > nuclei using 1.5 mg of protein of each preparation. The peroxidizability index was profoundly affected in polar lipids whereas that of neutral lipids was not. The most sensitive fatty acids for peroxidation were arachidonic acid, C20:4 n6 and docosahexanoic acid, C22:6 n3. Experiments carried out with membranes labelled in vivo indicate a selective release of radioactivity from polar rather than from neutral lipids during the peroxidation process. PMID- 9202434 TI - The use of Bacillus diarrhoeal enterotoxin (BDE) detection using an ELISA technique in the confirmation of the aetiology of Bacillus-mediated diarrhoea. AB - A commercially available ELISA kit was used for the detection of Bacillus diarrhoeal enterotoxin (BDE) in a variety of foods and faeces. The ability of isolates of Bacillus spp., including Bacillus cereus, to produce BDE in Brain Heart Infusion broth containing 0.1% glucose was also checked by use of the kit. Results show that 29 out of 31 B. cereus isolates were enterotoxigenic. Foods positive for performed BDE were always contaminated with > 10(5) B. cereus cfu g 1, but not all foods contaminated with large numbers of B. cereus were positive for BDE. Bacillus sp., other than one isolate which closely resembled B. subtilis, were negative for BDE production. Criteria for the confirmation of Bacillus-mediated diarrhoea should now include reports of symptoms and incubation periods consistent with the diarrhoeal form of food-poisoning by Bacillus spp., together with the results of tests for enterotoxigenicity of the Bacillus isolate, and detection of BDE in either the food and/or faeces. PMID- 9202435 TI - Mycobiota in Portuguese 'normal' and 'green' cork throughout the manufacturing process of stoppers. AB - The compounds responsible for the so-called 'cork taint' include, among others, some microbial metabolites which can be produced by the microbial population colonizing the unprocessed cork and stoppers. This study was intended to obtain information on the mycobiota associated with Portuguese cork throughout the manufacturing process of stoppers. Samples of barks and stoppers of both 'normal' and 'green' cork were examined. Moulds were isolated from 'normal' and 'green' cork throughout the entire cork stopper manufacturing process. Yeasts were rarely detected in the corks. Fungal contamination was not detected in finished stoppers from the company under study. PMID- 9202436 TI - Presence of additional peptidases in Streptococcus thermophilus CNRZ 302 compared to Lactococcus lactis. AB - Streptococcus thermophilus is widely used in the dairy industry but little is known about its peptidase system. The aim of this study was to determine the biochemical and genetic characteristics of this system, and to compare it to the well known system of Lactococcus lactis. We separated the intracellular proteins of Strep. thermophilus CNRZ 302 and L. lactis NCDO 763 by ion-exchange chromatography and we detected the activity of the different types of peptidases. In both L. lactis and Strep. thermophilus strains, we showed 13 different peptidase activities with biochemical homologies between both species. Streptococcus thermophilus also possessed two peptidases which we did not find in L. lactis: an aminopeptidase and an oligopeptidase. We performed Southern blot experiments and among the eight peptidase genes tested, only the genes encoding the general aminopeptidases, pepC and pepN, were homologous between the L. lactis and Strep. thermophilus strains. Besides biochemical and genetic similarities, the peptidase systems of Strep. thermophilus and L. lactis thus differed by the presence of additional peptidases in Strep. thermophilus. PMID- 9202437 TI - Factors contributing to the survival of poultry associated Pseudomonas spp. exposed to a quaternary ammonium compound. AB - Resistance to benzalkonium chloride (BC) among Pseudomonas spp. isolated from poultry carcasses was determined and strategies for elimination of resistant strains evaluated. This investigation showed that resistance was quite common, about 30% of the isolates being able to grow in 200 micrograms ml-1 BC. Pseudomonas fluorescens strains were generally less susceptible than strains of Ps. lundensis and Ps. fragi. An overnight incubation in medium containing 200 micrograms ml-1 BC was sufficient to reduce the susceptibility of two Pseudomonas strains to the lethal effect of BC significantly. Adding EDTA enhanced the lethal effect of BC, but the effect was reduced after growing cells in medium containing BC and EDTA. Growth in medium with a quaternary ammonium compound (QAC) rendered the cells more susceptible to chlorine, phenolics, and alkylaminoacetate. These results indicate that alternating use of QACs with these compounds can be used to avoid build-up of resistant strains. In addition, increased temperatures improved the lethal effect of BC and should be considered when planning disinfection routines. PMID- 9202438 TI - Restriction endonuclease analysis, DNA relatedness and phenotypic characterization of Campylobacter jejuni and Camp. coli isolates involved in food borne disease. AB - Seven cases of Campylobacter infection, each of them involving two isolates, were analysed. Study of their biochemical profiles and susceptibility patterns allowed the identification of Camp. jejuni and Camp. coli isolates and the effective typing of Camp. jejuni strains into biotypes. Genotyping was carried out by comparing chromosomal DNA restriction patterns obtained by cleavage with Bg/II and EcoRV and by Southern hybridization experiments. These studies revealed clonal homogeneity between both isolates in five of the seven cases studied, indicating that in these cases Campylobacter infection was caused by a single strain. Infection with two different strains was characterized in only two of the seven cases studied, two different species belonging to Camp. coli and Camp. jejuni ssp. jejuni biotype 1 being identified. Genetic analysis proved to be the most reliable technique to achieve precise identification of strains and to elucidate clonal heterogeneity among Campylobacter isolates obtained from a single patient. PMID- 9202439 TI - Effect of high pressure on the reduction of microbial populations in vegetables. AB - Resistance of micro-organisms to high pressure is variable and directly related to extrinsic and intrinsic factors. Pressures of 100, 200, 300, 350 and 400 MPa were applied at 20 degrees C for 10 min and at 10 degrees C for 20 min using strains of Gram-positive and Gram-negative bacteria, moulds and yeasts, as well as spores of Gram-positive bacteria. The results showed that at pressures of 100 and 200 MPa, decreases in microbial populations were not significant, whereas the populations of all the micro-organisms tested decreased considerably at a pressure of 300 MPa. A pressure of 300 MPa at 10 degrees C for 20 min was required to completely reduce the population of Saccharomyces cerevisiae, and a pressure of 350 MPa was needed to reduce most of the Gram-negative bacteria and moulds. The Gram-positive bacteria were more resistant, and pressures of 400 MPa were unable to completely reduce their populations. The different pressures employed had little effect on the initial numbers of spores. The initial populations of viable aerobic mesophiles and moulds and yeasts in vegetables (lettuce and tomatoes) decreased 1 log unit at pressures of 300 MPa and above under both sets of experimental treatment conditions. However, treatment at that pressure also resulted in alterations in the organoleptic properties of the samples. In the tomatoes, the skin loosened and peeled away, though the flesh remained firm, and colour and flavour were unchanged. The lettuce remained firm but underwent browning; flavour was unaffected. In vegetables use of moderate pressures in combination with other treatment conditions would appear to be required to reduce the populations of contaminating micro-organisms while avoiding the undesirable alterations in organoleptic properties that take place at 300 MPa. PMID- 9202440 TI - Survival of the recombinant Bacteroides thetaiotaomicron strain BTX in in vitro rumen incubations. AB - The survival of Bacteroides thetaiotaomicron strain BTX under rumen-simulating conditions was studied. Strain BTX is a recombinant variant of strain 5482 engineered for the production of high levels of xylanase, an enzyme important in the degradation of hemicellulose. Strain BTX was not inhibited by compounds present in rumen fluid and it grew well in media containing rumen fluid (up to 75%) or high concentrations of volatile fatty acids (total concentration, 100 mmol l-1). The ability of strain BTX to compete with other microorganisms under rumen-like conditions was studied in in vitro incubations of rumen contents. These experiments employed a consecutive batch culture (CBC) system consisting of alfalfa suspended in a rum flid buffer inoculated with blended rumen contents and maintained by transfers (10%, v/v) at 48 h intervals. CBC cultures contained a diversity of microbial morphotypes and accumulated fermentation products in rumen like proportions. WHen added alone, the numbers of BTX cells were maintained for only a few hours, and then declined precipitously until undetectable after 48 h. If CBC cultures were also supplemented with chondroitin sulphate (a mucopolysaccharide used by Bact. thetaiotaomicron), strain BTX grew and the pattern of its population generally followed that of the total population of ruminal bacteria in these cultures. When transferred into fresh CBC cultures containing chondroitin sulphate, BTX was again able to grow and increase in numbers, but to a diminished degree. Although BTX was able to survive and maintain itself in chondroitin sulphate supplemented cultures, this was at a very low level (10(5) ml-1). The potential for manipulation of rumen function by inoculation with recombinant bacteria is discussed. PMID- 9202441 TI - Bioactivity of selected plant essential oils against Listeria monocytogenes. AB - Ninety-three different commercial essential oils were screened for activity against 20 Listeria monocytogenes strains in vitro and the results correlated against the actual chemical composition of each oil. There was a substantial difference in the activity between different essential oils as expected, but there was also a difference in activity between different samples of the same essential oil. Strong anti-Listeria activity was often correlated with essential oils containing a high percentage of monoterpenes, eugenol, cinnamaldehyde, thymol, and sometimes with citronellol, limonene and geraniol. However, as there was often no correlation between the anti-Listeria activity and the main chemical components, it is possible that either there is a more complex relationship with the chemical composition (which includes the minor components) or that substantial adulteration had occurred in some essential oil samples. PMID- 9202442 TI - Efficacy of three prevention strategies against legionella in cooling water systems. AB - The efficacy of three different prevention strategies on legionella in cooling systems was studied. The strategies were as follows: (1) water temperature was lowered; (2) water quality was improved; or (3) the system as disinfected with polyhexamethylene biguanidechloride (PHMB) biocide or with 2-bromo-2-nitropropane 1,3-diol (BNPD) biocide. Lowering of water temperature was the most effective method to reduce the concentration of legionella in cooling systems. Improving of water quality resulted in a transitory disinfection effect. The additions of PHMB or BNPD decreased the concentrations of both legionella and heterotrophic bacteria in cooling water. The effect of biocides, however, lasted at the most only a few months. If possible, lowering water temperature and improving the water quality should be the primary practices for controlling bacterial growth in cooling systems. Regular biocide treatments should be incorporated into the maintenance procedures if technical improvements cannot be done or if their efficiency is too low. PMID- 9202443 TI - Fate of low temperature and acid-adapted Yersinia enterocolitica and Listeria monocytogenes that contaminate lactic acid decontaminated meat during chill storage. AB - Pathogens found in the environment of abattoirs may become adapted to lactic acid used to decontaminate meat. Such organisms are more acid tolerant than non adapted parents and can contaminate meat after lactic acid decontamination (LAD). The fate of acid-adapted Yersinia enterocolitica and Listeria monocytogenes, inoculated on skin surface of pork bellies 2 h after LAD, was examined during chilled storage. LAD included dipping in 1%, 2% or 5% lactic acid solutions at 55 degrees C for 120 s. LAD brought about sharp reductions in meat surface pH, but these recovered with time after LAD at approximately equal to 1-1.5 pH units below that of water-treated controls. Growth permitting pH at 4.8-5.2 was reached after 1% LAD in less than 0.5 d (pH 4.8-5.0), 2% LAD within 1.5 d (pH 4.9-5.1) and after 5% LAD (pH 5.0-5.2) within 4 d. During the lag on 2% LAD meat Y. enterocolitica counts decreased by 0.9 log10 cfu per cm2 and on 5% LAD the reduction was more than 1.4 log10 cfu per cm2. The reductions in L. monocytogenes were about a third of those in Y. enterocolitica. On 1% LAD the counts of both pathogens did not decrease significantly. The generation times of Y. enterocolitica and L. monocytogenes on 2-5% LAD meats were by up to twofold longer than on water-treated controls and on 1% LAD-treated meat they were similar to those on water-treated controls. Low temperature and acid-adapted L. monocytogenes and Y. enterocolitica that contaminate skin surface after hot 2-5% LAD did not cause an increased health hazard, although the number of Gram negative spoilage organisms were drastically reduced by hot 2-5% LAD and intrinsic (lactic acid content, pH) conditions were created that may benefit the survival and the growth of acid-adapted organisms. PMID- 9202444 TI - Research note: in vitro inhibition of Listeria monocytogenes growth by veillonellae cultures grown on tartrate media. AB - Veillonellae cultures were grown on agar media supplemented with tartrate and examined for inhibitory effects on the growth of Listeria monocytogenes. Veillonellae cultures grown on media supplemented with 0 or 50 mmol l-1 of tartrate did not inhibit the growth of L. monocytogenes; however, veillonellae grown on media supplemented with 100, 150 or 200 mmol l-1 of tartrate did inhibit the growth of L. monocytogenes. The inhibition of the growth of L. monocytogenes by the veillonellae was correlated with the increased production of acetate and propionate from tartrate by veillonellae and with the reduction of the pH of the media by L. monocytogenes. PMID- 9202445 TI - DNA probe and PCR-specific reaction for Lactobacillus plantarum. AB - A 300 bp DNA fragment of Lactobacillus plantarum isolated by randomly amplified polymorphic DNA (RAPD) analysis was cloned and sequenced. This fragment was tested using a dot-blot DNA hybridization to technique for its ability to identify Lact. plantarum strains. This probe hybridized with all Lact. plantarum strains tested and with some strains of Lact. pentosus, albeit more weakly. Two internal primers of this probe were selected (LbP11 and LbP12) and polymerase chain reaction (PCR) was carried out. All Lact. plantarum strains tested amplified a 250 bp fragment contrary to the other LAB species tested. This specific PCR for Lact. plantarum was also performed from colonies grown on MRS medium with similar results. These methods enabled the rapid and specific detection and identification of Lact. plantarum. PMID- 9202446 TI - Characterization of voltage- and Ca(2+)-activated K+ channels in squid olfactory receptor neurons. AB - We performed whole-cell voltage-clamp experiments on isolated olfactory neurons from the squid Lolliguncula brevis. Total outward currents were composed of three identifiable K+ currents: a delayed rectifier K+ current that showed slow inactivation and was sensitive to 5 mmol l-1 tetraethylammonium; a rapidly inactivating, 4-aminopyridine (4-AP)-sensitive, A-type K+ current and a Ca(2+) sensitive K+ current that was blocked by 200 nmol l-1 charybdotoxin and 10 mmol l 1 Cd2+ but was insensitive to apamin. The proportion of each current type varied from cell to cell, suggesting that responses to a given odorant would depend of the complement of channels present. The kinetics of the K+ currents were affected by temperature, with Q10 values ranging from 2 to 6. The identification and characterization of these K+ currents will greatly aid our understanding of action potential generation in these cells and will facilitate modelling of how odor responses are transduced and coded in squid olfactory receptor neurons. PMID- 9202447 TI - Brightness discrimination ability in the West Indian manatee (Trichechus manatus). AB - Two manatees were tested on their ability to discriminate brightness using a series of 30 shades of grey varying from white to black. The animals were trained to discriminate between different shades of grey in a twofold simultaneous-choice situation. Their ability to discern brightness differences correlates with Werber's law, and the calculated Werber fraction is 0.35. PMID- 9202448 TI - The role of Ca2+ in deflection-induced excitation of motile, mechanoresponsive balancer cilia in the ctenophore statocyst. AB - Motile, mechanoresponsive cilia (balancers) in ctenophore statocysts, like vertebrate hair cells, are excited or inhibited depending upon the direction in which they are deflected. Balancers, however, may become either excited (beat rapidly) or inhibited (beat slowly) by deflection in the same direction, depending on the sign of ctenophore geotaxis (positive or negative). The beat frequency of many cilia is controlled by concentrations of Ca2+, membrane potential and neural input. How these factors affect deflection-induced ciliary beating in balancers was investigated. Deflection-induced excitation of balancers in whole Mnemiopsis leidyi larvae and dissected adult (Mnemiopsis leidyi, Pleurobrachia pileus) statocysts was reversibly inhibited by the Ca2+ channel inhibitors Co2+, Mg2+, Ni2+, and Mn2+. Deflection-induced excitation in balancers of isolated adult M. leidyi balancer groups was also inhibited by Co2+ or by Ca(2+)-free medium. Isolated balancer group cilia, like balancer cilia of intact ctenophores, exhibited responses to either sign of geotaxis and graded responses to deflection. Isolated balancers that were chemically depolarized in high-[K+], Ca(2+)-free medium were excited by local application of Ca2+ onto the ciliary bases, but not onto the cell bases or the ciliary tips. It is proposed that deflection-induced excitation of balancers is due to influx of Ca2+ through stretch- and voltage-activated channel activity. The sign of geotaxis of whole larvae and dissected adult statocysts was switched by electrical stimulation. Thus, neural input may participate in reversing the directional sensitivity of balancer cells. PMID- 9202449 TI - Post-dive blood lactate concentrations in emperor penguins, Aptenodytes forsteri. AB - In order to determine an aerobic diving limit (ADL) in emperor penguins (Aptenodytes forsteri), post-dive blood lactate concentrations were measured in penguins foraging at an isolated sea ice hole. Resting lactate concentrations were 1.2-2.7 mmol l-1. Serial samples revealed that lactate level usually peaked within 5 min after dives and that 7-12 min was required for lactate concentrations to decrease from 5-8 mmol l-1 to less than 2.5 mmol l-1. Post-dive lactate level was not elevated above 3 mmol l-1 for dives shorter than 5 min. Two phase regression analysis revealed a transaction at 5.6 min in the post-dive lactate level versus diving duration relationship. All dives longer than 7 min were associated with lactate concentrations greater than 5 mmol l-1. We conclude that the ADL in emperor penguins ranges between 5 and 7 min. These are the first determinations of post-dive lactate concentrations in any free-diving bird and are currently the only physiological assessment of an ADL in an avian species. PMID- 9202450 TI - Design complexity and fracture control in the equine hoof wall. AB - Morphological and mechanical studies were conducted on samples of equine hoof wall to help elucidate the relationship between form and function of this complex, hierarchically organized structure. Morphological findings indicated a dependence of tubule size, shape and helical alignment of intermediate filaments (IFs) within the lamellae on the position through the wall thickness. The plane of the intertubular IFs changed from perpendicular to the tubule axis in the inner wall to almost parallel to the tubule axis in the outer wall. Morphological data predicted the existence of three crack diversion mechanisms which might prevent cracks from reaching the sensitive, living tissues of the hoof: a mid wall diversion mechanism of intertubular material to inhibit inward and upward crack propagation, and inner- and outer-wall diversion mechanisms that prevent inward crack propagation. Tensile and compact tension fracture tests were conducted on samples of fully hydrated equine hoof wall. Longitudinal stiffness decreased from 0.56 to 0.30 GPa proceeding inwardly, whereas ultimate (maximum) properties were constant. Fracture toughness parameters indicated that no compromise results from the declining stiffness, with J-integral values ranging from 5.5 to 7.8 kJ m-2 through the wall thickness; however, highest toughness was found in specimens with cracks initiated tangential to the wall surface (10.7 kJ m-2). Fracture paths agreed with morphological predictions and further suggested that the wall has evolved into a structure capable of both resisting and redirecting cracks initiated in numerous orientations. PMID- 9202451 TI - Direct measurement of flow from the posterior lymph hearts of hydrated and dehydrated toads (Bufo marinus). AB - Flow from the posterior lymph hearts of Bufo marinus was measured using Doppler flow probes. These probes were placed on the posterior vertebral vein and recorded flow as lymph was ejected from the heart. In resting, hydrated toads, mean lymph flow from one of the paired posterior lymph hearts was 25.9 +/- 4.9 ml kg-1 h-1, stroke volume was 8.9 +/- 1.4 microL kg-1 and lymph heart rate was 47.5 +/- 3.7 beats min-1. We estimate that, together, the paired posterior lymph hearts are capable of generating flows that are approximately one-sixtieth of the resting cardiac output. Mean peak systolic pressure developed by the posterior lymph hearts was 1.62 +/- 0.08 kPa. Simultaneous measurements of lymph heart pressure development and flow revealed that the outflow pore of the heart opened at a pressure of 0.71 +/- 0.04 kPa, approximately 113 +/- 5 ms into systole. When toads were moderately disturbed, stroke volume increased by as much as fourfold with little change in lymph heart rate (< 5 beats min-1). When toads were dehydrated, lymph flow decreased by 70% at 12h and by 80% and 24h. Since there was only a modest non-significant decrease in lymph heart rate (30%), this reduction in flow was attributed to decreases in stroke volume (approximately 80%). Lymph heart flow and stroke volume returned to control values 30 min after adding water back into the experimental chamber. Stroke volume was clearly more important in regulating lymph flow than lymph heart rate under these conditions in Bufo marinus. PMID- 9202452 TI - The effect of exercise-induced localised hyperthermia on tendon cell survival. AB - Tendons that store energy during locomotion, such as the equine superficial digital flexor tendon (SDFT) and human Achilles tendon, suffer a high incidence of central core degeneration which is thought to precede tendon rupture. Although energy storage contributes to the efficiency of locomotion, tendons are not perfectly elastic and some energy is lost in the form of heat. Recent studies have shown that the central core of equine SDFT reaches temperatures as high as 45 degrees C during high-speed locomotion. In this study, we test the hypothesis that hyperthermia causes tendon cell death and results in tendon central core degeneration. Tendon fibroblasts cultured from the core of the equine SDFT were subjected to a temperature of 45 degrees C in an in vitro system for 0-180 min, and cell survival fraction was measured and compared with that for equine dermal fibroblasts and a commercial rat kidney fibroblast cell line (NRK 49F). Tendon fibroblasts were significantly more resistant to hyperthermia than NRK 49F cells after 30, 45 and 60 min of heating and significantly more resistant than dermal fibroblasts after 45 and 60 min of heating. After 10 min of heating at 45 degrees C, the tendon fibroblast cell survival fraction was 91 +/- 4%, whereas heating for 10 min at 48 degrees C resulted in a drop in the cell survival fraction to 22 +/- 4%. In conclusion, while temperatures experienced in the central core of the SDFT in vivo are unlikely to result in tendon cell death, repeated hyperthermic insults may compromise cell metabolism of matrix components, resulting in tendon central core degeneration. PMID- 9202453 TI - Escherichia coli STb enterotoxin. PMID- 9202454 TI - Escherichia coli LT enterotoxin subunit A demonstrates partial toxicity independent of the nicking around Arg192. AB - A study was conducted into whether or not nicking of the A subunit of Escherichia coli LT enterotoxin at position Arg192 or its neighbouring amino acids Arg192 to The195 is required for its toxicity. The toxic activity of mutants created by substitution or deletion at this position, which lacked ADP-ribosyltransferase activity in vitro, was not completely obliterated and cyclic AMP was partially induced in the target cells, showing that they still displayed enzymic activity in vivo. Moreover, although the A subunit possesses three potential sites for cleavage by furin, furin was not involved in the partial toxicity and cyclic AMP induction observed. These data suggest that target cells have a nick mechanism that operates at sites other than those around Arg192 or those recognized by furin, which generates an active fragment by processing the A subunit after toxin binding to the cell membrane. PMID- 9202455 TI - VlpA of Vibrio cholerae O1: the first bacterial member of the alpha 2 microglobulin lipocalin superfamily. AB - We have identified a gene, vlpA, which is closely linked to the mfrA,B locus associated with mannose-fucose-resistant haemagglutination. VlpA is an outer membrane protein which can be labelled with [3H]palmitate and whose processing is globomycin-sensitive, suggesting that it is a lipoprotein. Homology searches revealed that VlpA belongs to the group of lipocalins of the alpha 2 microglobulin superfamily which function as small hydrophobic molecule transporters, and is the first identified bacterial member of this group. Multiple copies of this gene are present in Vibrio cholerae O1 and O139 and Southern hybridization reveals a biotype-specific pattern of fragment sizes. Construction of strains capable of hyperproducing VlpA suggested that it is able to bind haemin with low affinity but this may be due to a simple hydrophobic interaction. Attempts to construct specific mutants in vlpA have been unsuccessful, presumably because of the multiple copies of vlpA genes and their linkage to the VCR element. PMID- 9202456 TI - Targeted gene-replacement mutagenesis of dcrA, encoding an oxygen sensor of the sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough. AB - A gene-replacement mutagenesis method has been developed for the anaerobic, sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough and used to delete dcrA, encoding a potential oxygen or redox sensor with homology to the methyl-accepting chemotaxis proteins. A suicide plasmid, containing a cat-marked dcrA allele and a counter-selectable sacB marker was transferred from Escherichia coli S17-1 to D. vulgaris by conjugation. Following plasmid integration the desired dcrA deletion mutant (D. vulgaris F100) was obtained in media containing sucrose and chloramphenicol. Southern blot screening was required to distinguish D. vulgaris F100 from strain in which the sacB marker was inactivated by transposition of an endogenous IS element. No anaerotactic deficiency has so far been detected in D. vulgaris F100, which was found to be more resistant to inactivation by oxygen that the wild-type. Increased transcription of the rbo-rub operon, located immediately downstream from dcrA, was demonstrated by Northern blotting and may be the cause of this unusual phenotype, in view of the recent discovery that Rbo can complement the deleterious effects of superoxide dismutase deficiency in E. coli. PMID- 9202457 TI - Organization of methylamine utilization genes (mau) in 'Methylobacillus flagellatum ' KT and analysis of mau mutants. AB - The organization of genes involved in utilization of methylamine (mau genes) was studied in the obligate methylotroph 'Methylobacillus flagellatum' KT. Nine open reading frames were identified as corresponding to the genes mauFBEDAGLMN. In addition, an open reading frame (orf-1 encoding a polypeptide with unknown function was identified upstream of the mau gene cluster. Subclones of the 'M. flagellatum' KT gene cluster were used for complementation of a series of chemically induced mau mutants of 'M. flagellatum' KT. Mutants in mauF, mauB, mauE/D, mauA, mauG, mauL and mauM were identified. Two mutants (mau-18 and mau 19) were not complemented by the known mau genes. Since none of the chemically induced mutants studies had a defect of orf-1 or mauN, inserting mutants in these genes were constructed. Phenotypically the mutants fell into three groups. The mauF, mauB, mauE/D, mauA, mauG, mauL and mauM mutants do not grow on methylamine as a source of carbon and lack methylamine dehydrogenase activity, but they synthesize both the large and the small subunit polypeptides albeit at different ratios. The mau-18 and mau-19 mutants do not grow on methylamine as a source of carbon, and lack both methylamine dehydrogenase activity and the methylamine dehydrogenase subunits. The orf-1 and mauN mutants grow on methylamine as a source of carbon and synthesize wild-type levels of methylamine dehydrogenase. It has been shown earlier that the product of the mauM gene is not required for synthesis of active methylamine dehydrogenase in Methylobacterium extorquens AM1 and Paracoccus denitrificans. However, MauM is required for synthesis of functional methylamine dehydrogenase in 'M. flagellatum'. PMID- 9202458 TI - Construction and properties of aconitase mutants of Escherichia coli. AB - Escherichia coli contains two genes (acnA and acnB) encoding aconitase activities. An acnB mutant was engineered by replacing the chromosomal acnB gene by an internally deleted derivative containing a tetR cassette. An acnB double mutant was then made by transducing a previously constructed acnA::kanR mutation into the acnB::tetR strain. Western blotting confirmed that the AcnA and AcnB proteins were no longer produced by the corresponding mutants and PCR analysis showed that the chromosomal acnB gene had been replaced by the disrupted gene. Aerobic and anaerobic growth in glucose minimal medium were impaired but not abolished by the acnB mutation, indicating that the lesion is partially complemented by the acnA+ gene, and growth was enhanced by glutamate. The acnAB double mutant would not grow on unsupplemented glucose minimal medium and although it responded to glutamate like a typical auxotroph under anaerobic conditions, under aerobic conditions no response to glutamate was observed before it was over-grown by 'revertants' lacking citrate synthase (acnAB gltA). The acnAB double mutant retained a low but significant aconitase activity (< or = 5% of wild-type), designated AcnC. Enzymological and regulatory studies with acn lacZ fusions indicated that AcnB is the major aconitase, which is synthesized earlier in the growth cycle than AcnA, and subject to catabolite and anaerobic repression. PMID- 9202459 TI - Plasmid diversity in Chlamydia. AB - Chlamydiae exhibit low interspecies DNA homology and plasmids from different chlamydial species can be readily distinguished by Southern blot analysis and restriction enzyme profiling. In contrast, available plasmid sequence data from within the species Chlamydia trachomatis indicate that plasmids from human isolates are highly conserved. To evaluate the nature and extent of plasmid variation, the complete nucleotide sequences were determined for novel plasmids from three diverse non-human chlamydial isolates: pCpA1 from avian Chlamydia psittaci (N352); pCpnE1 from equine Chlamydia pneumoniae (N16); and pMoPn from C. trachomatis mouse pneumonitis. Comparison of the sequence data did not identify an overall biological function for the plasmid but did reveal considerable sequence conservation (> 60%) and a remarkably consistent genomic arrangement comprising eight major ORFs and four 22 bp tandem repeats. The plasmid sequences were close to 7500 nucleotides in length (pCpA1, 7553 bp; pMoPn, 7502 bp) however the equine C. pneumoniae plasmid was smaller (7362 bp) than all other chlamydial plasmids. The reduced size of this plasmid was due to a single large deletion occurring within ORF 1; this potentially generates two smaller ORFs. The disruption of ORF 1 is the only significant variation identified amongst the chlamydial plasmids and could prove important for future vector development studies. PMID- 9202460 TI - The Bacillus subtilis 168 chromosome from sspE to katA. AB - We have cloned and sequenced a 24.5 kb region of the Bacillus subtilis 168 chromosome spanning the sspE and katA genes. The region contains a ribosomal RNA operon, rrnD, a tRNA gene set, trnD and 17 ORFs, 16 with putative ribosome binding sites. Four of the ORFs (ORF2, ORF14, ORF16 and ORF17) match to known B. subtilis genes (sspE, thiA, senS and katA). Eight of the remaining ORF products show similarities with proteins present in the databases, including an ATP binding transport protein, a glutamate-1-semialdehyde aminotransferase, a thiol specific antioxidant protein, a mitomycin radical oxidase and a ferric uptake regulation protein. PMID- 9202461 TI - Nucleotide sequence and analysis of the phoB-rrnE-groESL region of the Bacillus subtilis chromosome. AB - A 36 kb sequence of the phoB-rrnE-groESL region of the Bacillus subtilis chromosome at around 55 degrees has been determined. The sequenced region contains 36 ORFs including the phoB and groESL genes, and the whole rrnE operon. The phoB gene is transcribed in the direction opposite to that of chromosome replication, while most ORFs, including groESL and the rrnE operon, are transcribed in the same direction. Two newly identified tRNA genes upstream of the rrnE operon were those for Arg-tRNA and Gly-tRNA. The sequenced region contains an operon consisting of genes for degradation and uptake of mannan. The rrnE operon and its downstream ORFs are well conserved among Mycoplasma genitalium, Haemophilus influenzae, Synechocystis sp. and Methanococcus jannaschii. delta H consensus sequences are present in the promoter regions of three ORFs, including groESL. PMID- 9202462 TI - Filamentous growth of the budding yeast Saccharomyces cerevisiae induced by overexpression of the WHi2 gene. AB - The WHI2 gene of the budding yeast Saccharomyces cerevisiae is required for the arrest of cell proliferation upon nutrient exhaustion: whi2 mutants carry on dividing and in the absence of growth become abnormally small. It is reported here that overexpression of Whi2 from the GAL1 promoter results in filamentous growth - cells fail to complete cytokinesis, the budding pattern changes from axial to polar, cells become elongated and cell size increases threefold. In many ways, these filaments resemble the pseudohyphae which result from nitrogen limited growth and the filaments seen during the invasive growth of haploids. However, Whi2-induced filament formation is reduced, but not blocked, by mutations in STE7, STE12 or STE20 which do block pseudohypha formation. Furthermore, pseudohypha formation can still occur in a diploid in which both copies of the WHI2 gene have been deleted. Thus Whi2-induced filament formation and pseudohypha formation must come about through the action of different pathways. Despite this, a mutation in the STE11 gene, which is required for pseudohypha formation, does block Whi2-induced filament formation. Concanavalin A pulse-chase experiments show that new cell wall material is incorporated only into the tips of the apical cells. An extragenic suppressor of the Whi2 allele also results in filamentous growth. These results suggest that Whi2 negatively regulates a function required for the budding mode of cell proliferation including cytokinesis. This function is defined wholly or in part by the fsw1 allele. PMID- 9202463 TI - Extrusion of benzoic acid in Saccharomyces cerevisiae by an energy-dependent mechanism. AB - When grown in the presence of benzoic acid, Saccharomyces cerevisiae was able to extrude [(14)C]benzoic acid when a pulse a glucose was given to preloaded cells. While octanoic, sorbic, hexanoic, salicylic, butyric and propionic acids were also inducers, ethanol and acetic acid were not. The mechanism of extrusion required energy and prior growth in the presence of the inducers. Diethylstilbestrol, an inhibitor of ATPases, prevented benzoic acid extrusion. Propionic acid was not actively extruded in cells adapted to either benzoic or propionic acid, behaving as an appropriate probe to measure intracellular pH. Even though the extrusion mechanism was active, benzoic acid entered the cells by a simple diffusion mechanism. PMID- 9202464 TI - An important role for glutathione and gamma-glutamyltranspeptidase in the supply of growth requirements during nitrogen starvation of the yeast Saccharomyces cerevisiae. AB - When the yeast Saccharomyces cerevisiae sigma 1278b was starved for nitrogen, the total glutathione (GSH) pool increased from 7 to 17 nmol (mg dry wt)-1 during the first 2 h and then declined. More than 90% of the total GSH shifted towards the central vacuole during this time. This transient stimulation was not observed in the presence of buthionine-(S,R)-sulphoximine (BSO), a specific transition-state analogue inhibitor of gamma-glutamylcysteine synthase (gamma-GCS), nor in a mutant strain deficient in this enzyme- gamma-Glutamyltranspeptidase (gamma-GT), a vacuolar enzyme responsible for the initial step of GSH degradation, was derepressed during nitrogen starvation. This mechanism can apparently enable the starved yeast cell to use the constituent amino acids from GSH which accumulate in the vacuole to satisfy its growth requirements for nitrogen. PMID- 9202465 TI - Effects of various types of stress on the metabolism of reserve carbohydrates in Saccharomyces cerevisiae: genetic evidence for a stress-induced recycling of glycogen and trehalose. AB - It is well known that glycogen and trehalose accumulate in yeast under nutrient starvation or entering into the stationary phase of growth, and that high levels of trehalose are found in heat-shocked cells. However, effects of various types of stress on trehalose, and especially on glycogen, are poorly documented. Taking into account that almost all genes encoding the enzymes involved in the metabolism of these two reserve carbohydrates contain between one and several copies of the stress-responsive element (STRE), an investigation was made of the possibility of a link between the potential transcriptional induction of these genes and the accumulation of glycogen and trehalose under different stress conditions. Using transcriptional fusions, it was found that all these genes were induced in a similar fashion, although to various extents, by temperature, osmotic and oxidative stresses. Experiments performed with an msn2/msn4 double mutant proved that the transcriptional induction of the genes encoding glycogen synthase (GSY2) and trehalose-6-phosphate synthase (TPS1) was needed for the small increase in glycogen and trehalose upon exposure to a mild heat stress and salt shock. However, the extent of transcriptional activation of these genes upon stresses in wild-type strains was not correlated with a proportional rise in either glycogen or trehalose. The major explanation for this lack of correlation comes from the fact that genes encoding the enzymes of the biosynthetic and of the biodegradative pathways were almost equally induced. Hence, trehalose and glycogen accumulated to much higher levels in cells lacking neutral trehalose or glycogen phosphorylase exposed to stress conditions, which suggested that one of the major effects of stress in yeast is to induce a wasteful expenditure of energy by increasing the recycling of these molecules. We also found that transcriptional induction of STRE-controlled genes was abolished at temperatures above 40 degree C, while induction was still observed for a heat-shock-element regulated gene. Remarkably, trehalose accumulated to very high levels under this condition. This can be explained by a stimulation of trehalose synthase and inhibition of trehalose by high temperature. PMID- 9202467 TI - The relationship between external glucose concentration and cAMP levels inside Escherichia coli: implications for models of phosphotransferase-mediated regulation of adenylate cyclase. AB - The concentration of glucose in the medium influences the regulation of cAMP levels in Escherichia coli. Growth in minimal medium with micromolar glucose results in 8- to 10-fold higher intracellular cAMP concentrations than observed during growth with excess glucose. Current models would suggest that the difference in cAMP levels between glucose-rich and glucose-limited states is due to altered transport flux through the phosphoenolpyruvate: glucose phosphotransferase system (PTS), which in turn controls adenylate cyclase. A consequence of this model is that cAMP levels should be inversely related to the saturation of the PTS transporter. To test this hypothesis, the relationship between external glucose concentration and cAMP levels inside E. coli were investigated in detail, both through direct cAMP assay and indirectly through measurement of expression of cAMP-regulated genes. Responses were followed in batch, dialysis and glucose-limited continuous culture. A sharp rise in intracellular cAMP occurred when the nutrient concentration in minimal medium dropped to approximately 0.3 mM glucose. Likewise, addition of > 0.3 mM glucose, but not < 0.3 mM glucose, sharply reduced the intracellular cAMP level of starving bacteria. There was no striking shift in growth rate or [14C] glucose assimilation in bacteria passing through the 0.5 to 0.3 mM concentration threshold influencing cAMP levels, suggesting that neither metabolic flux nor transporter saturation influenced the sensing of nutrient levels. The (IIA/IIBC)Glc PTS is 96-97% saturated at 0.3 mM glucose so these results are not easily reconcilable with current models of cAMP regulation. Aside from the transition in cAMP levels initiated above 0.3 mM, a second shift occurred below 1 muM glucose. Approaching starvation, well below saturation of the PTS, cAMP levels either increased or decreased depending on unknown factors that differ between common E. coli K-12 strains. PMID- 9202466 TI - Glucose-dependent, cAMP-mediated ATP efflux from Saccharomyces cerevisiae. AB - Extracellular ATP plays an important role in the physiology of multicellular organisms; however, it is unknown whether unicellular organisms such as yeast also release ATP extracellularly. Experiments are described here which show that Saccharomyces cerevisiae releases ATP to the extracellular fluid. This efflux required glucose and the rate was increased dramatically by the proton ionophores nigericin, monensin, carbonyl cyanide m-chlorophenylhydrazone and carbonyl cyanide p-(trifluoromethoxy)-phenylhydrazone; ATP efflux was also increased by the plasma membrane proton pump inhibitor diethylstilbestrol. The increase in the concentration of extracellular ATP was not due to cell lysis or general disruption of plasma membrane integrity as measured by colony-forming and methylene-blue-staining assays. ATP efflux was strictly correlated with a rise in intracellular cAMP; therefore, the cAMP pathway is likely to be involved in triggering ATP efflux. These results demonstrate that yeast cells release ATP in a regulated manner. PMID- 9202468 TI - Anaerobic taurine oxidation: a novel reaction by a nitrate-reducing Alcaligenes sp. AB - Enrichment cultures were prepared under strictly anoxic conditions in medium representing fresh water and containing an organosulfonate as electron donor and carbon source, and nitrate as electron acceptor. The inoculum was from the anaerobic digestor of two communal sewage works. The natural organosulfonates 2 aminoethanesulfonate (taurine), DL-2-amino-3-sulfopropionate (cysteate) and 2 hydroxyethanesulfonate (isethionate) all gave positive enrichments, whereas unsubstituted alkanesulfonates, such as methanesulfonate and arenesulfonates, gave no enrichment. Two representative enrichments were used to obtain pure cultures, and strains NKNTAU (utilizing taurine) and NKNIS (utilizing isethionate) were isolated. Strain NKNTAU was examined in detail. Out of 18 tested organosulfonates, it utilized only one, taurine, and was identified as a novel Alcaligenes sp., a facultatively anaerobic bacterium. Carbon from taurine was converted to cell material and carbon dioxide. The amino group was released as ammonium ion and the sulfonate moiety was recovered as sulfate. Nitrate was reduced to nitrogen gas. PMID- 9202469 TI - Xanthobacter flavus employs a single triosephosphate isomerase for heterotrophic and autotrophic metabolism. AB - The expression of the cbb and gap-pgk operons of Xanthobacter flavus encoding enzymes of the Calvin cycle is regulated by the transcriptional regulator CbbR. In order to identify other genes involved in the regulation of these operons, a mutant was isolated with a lowered activity of a fusion between the promoter of the cbb operon and the reporter gene lacZ. This mutant was unable to grow autotrophically and had a reduced growth rate on medium supplemented with gluconate or succinate. The regulation of the gap-pgk operon in the mutant was indistinguishable from the wild-type strain, but induction of the cbb operon upon transition to autotrophic growth conditions was delayed. Complementation of the mutant with a genomic library of X. flavus resulted in the isolation of a 1.1 kb ApaI fragment which restored autotrophic growth of the mutant. One open reading frame (ORF) was present on the ApaI fragment, which could encode a protein highly similar to triosephosphate isomerase proteins from other bacteria. Cell extracts of the mutant grown under glycolytic or gluconeogenic conditions had severely reduced triosephosphate isomerase activities. The ORF was therefore identified as tpi, encoding triosephosphate isomerase. The tpi gene is not linked to the previously identified operons encoding Calvin cycle enzymes and therefore represents a third transcriptional unit required for autotrophic metabolism. PMID- 9202470 TI - The thioredoxin reductase system of mycoplasmas. AB - Representative species of the Mollicutes possess a thioredoxin reductase system (NTS) composed of a low-molecular-mass thioredoxin (TRX) and NADPH-binding thioredoxin reductase (NTR). The TRXs of Mycoplasma pneumoniae and M. capricolum have molecular masses of 11-2 and 12 kDa, respectively, and are stable at 90 degree C for 10 min. Both TRXs reacted with monospecific polyclonal antibodies generated against the Bacillus subtilis TRX, but not with anti-Escherichia coli TRX antisera. The M. capricolum and M. pneumoniae NTRs were partially purified and were found to be active with the homologous TRX, but not with the TRX of B. subtilis or E. coli. The NTS activity had an optimal pH of 6.5-7.5 and was dependent on NADPH as an election donor, a requirement which could not be fulfilled by NADH. The genes encoding the TRX and NTR (trxA and trxB) or M. pneumoniae were cloned and sequenced. The comparative analysis of the predicted amino acid sequence of trxA showed that the 11.2 kDa protein (102 aa) shared 26 68% sequence similarity with products of other known trxA genes and contained the conserved active site Cys-Gly-Pro-Cys. The predicted amino acid sequence of trxB contained 315 residues with a conserved NADPH binding domain and FAD binding domains I and II. The cysteine dithiol redox active region had isoleucine rather than threonine at the active site, as compared with other NTRs. The high activity of the NTS in mycoplasmas suggests that mycoplasmas may have evolved the NTS to protect themselves from the consequences of their self-generated oxidative challenge. PMID- 9202471 TI - Regulation of exopolysaccharide production in Rhizobium leguminosarum biovar viciae WSM710 involves exoR. AB - A mildly acid-sensitive mutant of Rhizobium leguminosarum bv. viciae WSM710 (WR6 35) produced colonies which were more mucoid in phenotype than the wild-type. Strain WR6-35 contained a single copy of Tn5 and the observed mucoid phenotype, acid sensitivity and Tn5-induced kanamycin resistance were 100% co-transducible using phage RL38. WR6-35 produced threefold more exopolysaccharide (EPS) than the wild-type in minimal medium devoid of a nitrogen source. EPS produced by the mutant and the wild-type was identical as determined by proton NMR spectra. An EcoRI rhizobial fragment containing Tn5 and flanking rhizobial sequences was cloned from the mutant, restriction mapped and sequenced. There was extensive similarity between the ORF disrupted by Tn5 in R. leguminosarum bv. viciae WR6-35 and the exoR gene of Rhizobium (Sinorhizobium) meliloti Rm1021 (71.3% identity over 892 bp). At the protein level there was 70% identity and 93.3% similarity over 267 amino acids with the ExoR protein of R. meliloti Rm1021. Hydrophilicity profiles of the two proteins from these two rhizobia are superimposable. This gene in R. leguminosarum bv. viciae was thus designated exoR. The data suggest that Tn5 has disrupted a regulatory gene encoding a protein that negatively modulates EPS biosynthesis in R. leguminosarum bv. viciae WSM710. Despite earlier suggestions that EPS production and acid tolerance might be positively correlated, disruption of exoR in either R. leguminosarum bv. viciae or R. meliloti and its associated overproduction of EPS does not result in a more acid tolerant phenotype than the wild-type when cultures are screened on conventional laboratory agar. PMID- 9202472 TI - A Neurospora crassa mutant altered in the regulation of L-amino acid oxidase. AB - The isolation and characterization of a Neurospora crassa mutant altered in L amino oxidase regulation is reported. The previously isolated gln-1bR8 strain, which only synthesizes the glutamine synthetase alpha monomer and lacks the beta monomer, was used as parental strain. A mutant derivative of strain was selected for its ability to grow on minimal medium in the presence of DL-methionine-SR sulfoximine (MSO), an inhibitor of glutamine synthetase activity. This gln 1bR8;MSOR mutant overcame the inhibitory effect of MSO by increasing the activity of L-amino acid oxidase, an enzyme capable of degrading this compound. In contrast with the wild-type strain, the L-amino acid oxidase of the MSOR mutant was resistant to glutamine repression; in fact, it was induced by this amino acid but repressed by ammonium. This mutant is different from other nitrogen regulatory N. crassa mutants reported and is only altered in the regulation of L amino acid oxidase. The MSOR mutation is epistatic to nit-2 since the nit2;MSOR double mutant regulated the L-amino acid oxidase in the same way as the MSOR single mutant. PMID- 9202473 TI - Identification of a phenolic 3-O-methyltransferase in the lignin-degrading fungus Phanerochaete chrysosporium. AB - A methyltransferase enzyme catalysing the 3-O-methylation of isovanillic acid (3 hydroxy-4-methoxybenzoic acid) by S-adenosylmethionine (SAM) was identified in Phanerochaete chrysosporium and purified. Gel filtration indicated an M(r) of 71,000 and SDS-PAGE showed that the enzyme was composed of two subunits of M(r) approximately 36,000. Substrate utilization studies demonstrated that the enzyme was highly specific, displaying an exclusive preference for the methylation of the 3-hydroxyl group of several substituted benzoic acids. 3-Hydroxybenzoic acids with a methoxyl or hydroxyl substituent in the 2 or 4 position were the best substrates with isovanillic and 3,4-dihydroxybenzoic acids showing the highest rates of methylation. The 3-O-methyltransferase enzyme was induced later in the growth cycle than the 4-O-methyltransferase previously isolated from this fungus, which is believed to have a role in the 4-O-methylation of lignin degradation products. However the function of this meta-specific enzyme, the first phenolic 3 O-methyltransferase isolated from a fungus, remains unclear. The combined activities of the 3- and 4-O-methyltransferase enzymes satisfactorily account for the pattern of SAM-dependent methylating activity shown by whole mycelia to phenolic substrates. PMID- 9202474 TI - Adaptation of proteases and carbohydrates of saprophytic, phytopathogenic and entomopathogenic fungi to the requirements of their ecological niches. AB - The abilities of isolates of saprophytes (Neurospora crassa, Aspergillus nidulans), an opportunistic human pathogen (Aspergillus fumigatus), an opportunistic insect pathogen (Aspergillus flavus), plant pathogens (Verticillium albo-atrum, Verticillium dahliae, Nectria haematococca), a mushroom pathogen (Verticillium fungicola) and entomopathogens (Verticillium lecanii, Beauveria bassiana, Metarhizium anisopliae) to utilize plant cell walls and insect cuticle components in different nutrient media were compared. The pathogens showed enzymic adaptation to the polymers present in the integuments of their particular hosts. Thus, the plant pathogens produced high levels of enzymes capable of degrading pectic polysaccharides, cellulose and xylan, as well as cutinase substrate, but secreted little or no chitinase and showed no proteolytic activity against elastin and mucin. The entomopathogens and V. fungicola degraded a broad spectrum of proteins (including elastin and mucin) but, except for chitinase, cellulase (V. lecanii and V. fungicola only) and cutinase (B. bassiana only), produced very low levels of polysaccharidases. The saprophytes (Neu. crassa and A. nidulans) and the opportunistic pathogens (A. fumigatus and A. flavus) produced the broadest spectrum of protein and polysaccharide degrading enzymes, indicative of their less specialized nutritional status. V. lecanii and V. albo atrum were compared in more detail to identity factors that distinguish plant and insect pathogens. V. albo-atrum, but not V. lecanii, grew well on different plant cell wall components. The major class of proteases produced in different media by isolates of V. albo-atrum and V. dahliae were broad spectrum basic (pI > 10) trypsins which degrade Z-AA-AA-Arg-NA substrates (Z, benzoyl; AA, various amino acids; Na, nitroanilide), hide protein azure and insect (Manduca sexta) cuticles. Analogous peptidases were produced by isolates of V. lecanii and V. fungicola but they were specific for Z-Phe-Val-Arg-NA. V. albo-atrum and V. dahliae also produced low levels of neutral (pI ca 7) and basic (pI ca 9.5) subtilisin-like proteases active against a chymotrypsin substrate (Succinyl-Ala2-Pro-Phe-NA) and insect cuticle. In contrast, subtilisins comprised the major protease component secreted by V. lecanii and V. fungicola. Both V. lecanii and V. albo-atrum produced the highest levels of subtilisin and trypsin-like activities during growth on collagen or insect cuticle. Results are discussed in terms of the adaptation of fungi to the requirements of their ecological niches. PMID- 9202475 TI - Isolation of developmentally regulated genes from the edible mushroom Agaricus bisporus. AB - From a cDNA library, constructed from mushroom primordia, nine cDNAs were isolated which were either induced or specifically expressed during fruit body development and maturation of the basidiomycete Agaricus bisporus. These cDNAs varied in size from 372 to 1019 bp and hybridized to transcripts of 400-1600 nt. Four of the cDNAs were only expressed in the generative phase of the life cycle while the other five cDNAs were strongly induced but had low steady-state mRNA levels in vegetatively grown mycelium of the hybrid strain Horst U1. An apparent full-length cDNA could be identified by sequence analysis and specified a putative protein homologous to the delta-subunit of the mitochondrial ATP synthase complex of Saccharomyces cerevisiae and Neurospora crassa. For one of the partial cDNAs, significant homology was found with a family of cell division control proteins, while another partial cDNA appeared to encode a cytochrome P450. All cDNAs, except the presumed cytochrome-P450-specifying cDNA (cypA), hybridized with single copy genes scattered over the Agaricus genome. For the cypA gene, the presence of several additional copies was shown by heterologous hybridizations. Based on changes in expression levels of the fruit-body-induced genes during development coinciding with alterations in morphological appearance of mushrooms, four stages of development were distinguished during growth and maturation of A. bisporus fruit bodies. PMID- 9202476 TI - beta-Tubulin genes and the basis for benzimidazole sensitivity of the opportunistic fungus Cryptococcus neoformans. AB - The basidiomycete Cryptococcus neoformans causes life-threatening infections in immunocompromised patients, and available chemotherapeutic agents are potentially toxic or have limited efficacy. In vitro, C. neoformans is very sensitive to selected benzimidazole compounds (e.g. albendazole), which act by disrupting microtubules through binding to the beta-tubulin subunit. To understand the basis for this benzimidazole sensitivity, we have characterized C. neoformans beta tubulin genes and their expression. Analysis of PCR amplification products, genomic and cDNA clones and Southern blots identified two beta-tubulin genes. TUB1 contains seven introns, including one that splits the start codon, and encodes a 447 amino acid protein with > 80% identity to most other beta-tubulins. A partial sequence of TUB2 revealed a higher density of introns and a considerably more divergent beta-tubulin. The relative expression of TUB1 to TUB2 determined by reverse-transcription PCR was about 3:1, consistent with a more limited role for the TUB2 product. Comparisons of beta-tubulin sequences from C. neoformans and from various benzimidazole-sensitive and -resistant organisms strongly suggest that the TUB1 product represents the primary benzimidazole target. This was supported by the identification of a His6 to Gln change in TUB1 from three independently isolated albendazole-resistant mutants. PMID- 9202477 TI - CHS2, a chitin synthase gene from the oomycete Saprolegnia monoica. AB - PCR was used to amplify fragments corresponding to the chitin synthase (CHS) genes from the Oomycetes Saprolegnia monoica, Phytophthora capsici and Achlya ambisexualis, utilizing as primers, oligonucleotides designed from the conserved region of CHS genes of chitinous fungi. Chitin synthase homologues were found in the three cellulosic fungi. The chitin synthase 2 gene (CHS2) from S. monoica was cloned, sequenced and characterized. The amino acid sequence deduced from the CHS2 genomic DNA revealed several domains, corresponding to the catalytic domains and polypeptide signatures, of high identity with CHS genes from chitinous fungi. Existence of a CHS gene family in S. monoica was supported by the identification of two CHS sequences among the PCR products, the localization of CHS homologues on two chromosomes, and the detection of two transcripts in mycelia and protoplasts. Polyclonal anti-chitin synthase antibodies raised against the N terminal and the neutral fragments of the CHS2 products revealed, respectively, two and four proteins in membrane fractions and a truncated active form in entrapped product. The overall comparison of the structure and organization of CHS genes indicates that in spite of their divergent evolution, Oomycetes and chitinous fungi have evolved with conserved chitin synthase systems. PMID- 9202479 TI - The genetic structure of Escherichia coli populations in feral house mice. AB - Escherichia coli was isolated from feral house mice (Mus domesticus) during the course of a mouse plague in the state of Victoria, Australia. Two farms were sampled over a period of 7 months and a total of 447 isolates were collected. The isolates were characterized using the techniques of randomly amplified polymorphic DNA and multi-locus enzyme electrophoresis. The mean genetic diversity of this E. coli population (H = 0.24) was found to be substantially lower than the diversity of an E. col population reported elsewhere for a single human host. Analysis of the allozyme data revealed that there were significant differences in the relative abundance of genotypes between the two localities sampled and among sample dates. Overall, however, spatial and temporal effects accounted for less than 5% of the genotypic diversity. Allele frequencies and the relative abundance of the more common genotypes did not differ between male and female hosts. The number of genotypes and genotype diversity increased as the age of the host increased, suggesting that the mice are continuing to acquire new E. coli clones throughout their life. The frequency of some alleles changed with respect to host age, which indicates that clone acquisition may not be a random process. It is argued that the low level of genetic diversity observed in this population of E. coli reflects the boom and bust nature of mouse population density in this region of Australia. PMID- 9202478 TI - Authentic display of a cholera toxin epitope by chimeric type 1 fimbriae: effects of insert position and host background. AB - The potential of the major structural protein of type 1 fimbriae as a display system for heterologous sequences was tested. As a reporter-epitope, a heterologous sequence mimicking a neutralizing epitope of the cholera toxin B chain was inserted, in one or two copies, into four different positions in the fimA gene. This was carried out by introduction of new restriction sites by PCR mediated site-directed mutagenesis of fimA in positions predicted to correspond to optimally surface-located regions of the subunit protein. Subsequently, the synthetic cholera-toxin-encoding DNA segment was inserted. Several of the chosen positions seemed amenable even for large foreign inserts; the chimeric proteins were exposed on the bacterial surface and the cholera toxin epitope was authentically displayed, i.e. it was recognized on bacteria by specific antiserum. Display of chimeric fimbriae was tested with respect to host background in three different Escherichia coli strains, i.e. an isogenic set of K 12 strains, differing in the presence of an indigenous fim gene cluster, as well as a wild-type isolate. Immunization of rabbits with purified chimeric fimbriae resulted in serum which specifically recognized cholera toxin B chain, confirming the utility of the employed strategy. PMID- 9202480 TI - Interruption of the Streptococcus gordonii M5 sspA/sspB intergenic region by an insertion sequence related to IS1167 of Streptococcus pneumoniae. AB - Streptococcus gordonii M5 and DL1 each express two related adhesin polypeptides, SspA and SspB, which are members of the antigen I/II family of streptococcal surface proteins. The sspA and sspB genes are tandemly arranged in both strains, with sspA residing upstream of sspB. The genes are separated by approximately 400 nucleotides in S. gordonii DL1 and 1300 nucleotides in S. gordonii M5. The nucleotide sequence of the sspA/sspB intergenic region of strain M5 is reported and the difference in length compared to S. gordonii DL1 shown to arise from the presence of an insertion sequence, designated ISSg1, consisting of 1197 bp. The nucleotide sequence of ISSg1 is highly homologous to IS1167 to Streptococcus pneumoniae and is related to a lesser extent to other members of the IS1096 family of bacterial insertion sequences. It contains a single ORF of 1026 bp, encoding a putative transposase polypeptide of 342 amino acids. The deduced transposase sequence exhibits 93% identity with the transposase polypeptides encoded by IS1167. However, the S. gordonii protein lacks a 90 residue central domain that is present in the IS1167 transposase and in the transposase polypeptides encoded by the related IS elements. In addition, the organization of the inverted repeats flanking the transposase gene in S. gordonii differs from IS1167. Extension products generated from a sspB-specific primer indicated that transcription initiates within the intergenic region in both S-gordonii strains, suggesting that sspA and sspB are independently transcribed. Transcription appears to initiate 42 bases upstream of sspB in S. gordonii DL1. In contrast, sspB transcription in M5 initiates at least 125 bases upstream of sspB, in close proximity to the terminal inverted repeat of ISsg1. These results indicate that the sspB promoter of S. gordonii M5 and DL1 are not conserved and suggest that ISSg1 sequences may play a role in directing the expression of sspB in S. gordonii M5. PMID- 9202481 TI - The site-specific recombinase encoded by pinD in Shigella dysenteriae is due to the presence of a defective Mu prophage. AB - The DNA inversion systems are made up of an invertible DNA segment and a site specific recombinase gene. Five systems are known in prokaryotes: the Salmonella typhimurium H segment and hin gene (H-hin), phage Mu G-gin, phage P1 C-cin, Escherichia coli e14 P-pin, and Shigella sonnei B-pinB systems. In this report a site-specific recombinase (pinD) gene of Shigella dysenteriae was cloned and sequenced. pinD mediated inversion of five known segments at the same extent in E. coli. Although one inv sequence was identified, no invertible region was detected in a cloned fragment. The predicted amino acid sequences of PinD and three ORFs showed high homology to those of Gin and its flanking gene products. An ORF homologous to Mom of Mu conserved a functional activity to modify intracellular plasmid DNA. Southern analysis showed that the cloned fragment contains two homologous regions corresponding to the left and right ends of the Mu genome. Together these results indicated that the pinD gene in S. dysenteriae is derived from a Mu-like prophage. PMID- 9202482 TI - Modulation of gene expression through chromosomal positioning in Escherichia coli. AB - Variations in expression of the nah genes of the NAH7 (naphthalene biodegradation) plasmid of Pseudomonas putida when placed in different chromosomal locations in Escherichia coli have been studied by employing a collection of hybrid mini-T5 transposons bearing lacZ fusions to the Psal promoter, along with the cognate regulatory gene nahR. Insertions of Psal-lacZ reporters in the proximity of the chromosomal origin of replication, oriC, increased accumulation of beta-galactosidase in vivo. Position-dependent changes in expression of the reporter product could not be associated with local variations of the supercoiling in the DNA region, as revealed by probing the chromosome with mobile gyrB-lacZ elements. Such variations in beta-galactosidase activity (and, therefore, the expression of catabolic genes) seemed, instead, to be linked to the increase in gene dosage associated with regions close to oriC, and not to local variations in chromosome structure. The tolerance of strains to the selection markers borne by the transposons also varied in parallel with the changes in LacZ levels. The role of chromosomal positioning as a mechanism for the outcome of adaptation phenotypes is discussed. PMID- 9202483 TI - Metabolic regulation of lrp gene expression in Escherichia coli K-12. AB - Expression of the lrp gene is regulated in part by the nutrients available to the cell, and is decreased in rich medium, in glucose minimal media enriched with amino acids, and in minimal medium with alternative carbon sources, such as acetate and succinate. When Lrp production is increased in a given medium, expression of its target genes is also increased. However, when the medium is changed from glucose to acetate, the response of the target genes is governed by many factors. PMID- 9202486 TI - Air pollution and human health: perspectives for the '90s and beyond. AB - This paper considers the health effects of air pollution from three perspectives: historical, statistical, and public policy, and also as depicted by the recent epidemiology, primarily mortality studies. The historical perspectives establish the reality of population-based health effects, and they provide data with which to evaluate more recent evidence. Statistical perspectives imply that, while there is strong evidence that associations between air quality and health persist, many details of these relationships remain obscure, especially as to the existence of concentration thresholds that might define safe exposure levels. Additional major questions include the effects of uncertainties in actual pollution exposures, the degree of prematurity of "excess" deaths, and whether the development of new cases of chronic disease is associated with air pollution. Public policy issues center around interpreting the new epidemiological studies in the light of these uncertainties and the analysis and management of the concomitant health risks. PMID- 9202484 TI - The aldA gene of Escherichia coli is under the control of at least three transcriptional regulators. AB - Expression studies on the aldA gene encoding aldehyde dehydrogenase in Escherichia coli showed induction by two types of molecule (hydroxyaldehydes and 2-oxoglutarate), carbon catabolite repression and respiration dependence. Promoter deletion analysis showed that the proximal operator, which includes inducer-regulator complex and catabolite repression protein (Crp) recognition sites, was necessary for induction by either type of inducer, and that full induction by aldehydes required the cooperation of distal operator sequences beyond position -119. Interactions of the regulator protein with the -59 to -6 fragment were shown by DNA mobility shift assays. Fusions of different deletions of the aldA promoter to lacZ indicated that a Crp site proximal to the transcriptional start point (tsp) was functional in the cAMP-dependent catabolite repression of this system, whereas a distal control site was likely to operate in a cAMP-independent catabolite repression. DNA mobility shift and footprint analyses showed that only the tsp proximal site was bound by pure Crp with a Kd of 5.4 x 10(-7) M. As shown by an Arc-defective strain, the aldA gene seems to be repressed by the Arc system under anaerobiosis, displaying its physiological full induction and activity in the presence of oxygen. PMID- 9202487 TI - Neighborhood quality, environmental hazards, personality traits, and resident actions. AB - A survey of 798 New Jersey residents examined relationships among residents' neighborhood activities, perceptions of neighborhood quality, trust of experts, support for rebuilding cities and equal rights, and degree of optimism. Neighborhood activities increased with lack of trust and optimism. These personality characteristic measures were folded into multidimensional constructs that included local environmental hazards, respondents' ratings of their previous neighborhoods, and some demographic variables. Pessimism and values that support equal rights and rebuilding cities were weakly associated with poor quality neighborhood ratings. PMID- 9202488 TI - Multimedia benchmarking analysis for three risk assessment models: RESRAD, MMSOILS, and MEPAS. AB - This paper is one in a series that describes results of a benchmarking analysis initiated by the Department of Energy (DOE) and the United States Environmental Protection Agency (EPA). An overview of the study is provided in a companion paper by Laniak et al. presented in this journal issue. The three models used in the study--RESRAD (DOE), MMSOILS (EPA), and MEPAS (DOE)--represent analytically based tools that are used by the respective agencies for performing human exposure and health risk assessments. Both single media and multimedia benchmarking scenarios were developed and executed. In this paper, the multimedia scenario is examined. That scenario consists of a hypothetical landfill that initially contained uranium-238 and methylene chloride. The multimedia models predict the fate of these contaminants, plus the progeny of uranium-238, through the unsaturated zone, saturated zone, surface water, and atmosphere. Carcinogenic risks are calculated from exposure to the contaminants via multiple pathways. Results of the tests show that differences in model endpoint estimates arise from both differences in the models' mathematical formulations and assumptions related to the implementation of the scenarios. PMID- 9202489 TI - An overview of a multimedia benchmarking analysis for three risk assessment models: RESRAD, MMSOILS, and MEPAS. AB - Multimedia modelers from the United States Environmental Protection Agency (EPA) and the United States Department of Energy (DOE) collaborated to conduct a detailed and quantitative benchmarking analysis of three multimedia models. The three models--RESRAD (DOE), MMSOILS (EPA), and MEPAS (DOE)--represent analytically-based tools that are used by the respective agencies for performing human exposure and health risk assessments. The study is performed by individuals who participate directly in the ongoing design, development, and application of the models. Model form and function are compared by applying the models to a series of hypothetical problems, first isolating individual modules (e.g., atmospheric, surface water, groundwater) and then simulating multimedia-based risk resulting from contaminant release from a single source to multiple environmental media. Study results show that the models differ with respect to environmental processes included (i.e., model features) and the mathematical formulation and assumptions related to the implementation of solutions. Depending on the application, numerical estimates resulting from the models may vary over several orders-of-magnitude. On the other hand, two or more differences may offset each other such that model predictions are virtually equal. The conclusion from these results is that multimedia models are complex due to the integration of the many components of a risk assessment and this complexity must be fully appreciated during each step of the modeling process (i.e., model selection, problem conceptualization, model application, and interpretation of results). PMID- 9202490 TI - Critical effects and exposure limits. AB - The use of critical effects in the determination of occupational exposure limits (OELs) in Sweden is subjected to a statistical study. Many of the present OELs are high in relation to known no-effect levels and effect levels, and the degree of protection has a surprisingly weak correlation with the seriousness of the adverse effect. Several proposals for improved procedures are put forward. One of these is to supplement the concept of critical effects with that of dominant effects. A dominant effect of a substance is a health effect that is at some concentration the most serious health effect. PMID- 9202491 TI - Molecular physiology of plant sulfur metabolism. PMID- 9202492 TI - Extensin from suspension-cultured potato cells: a hydroxyproline-rich glycoprotein, devoid of agglutinin activity. AB - Enhanced deposition and cross-linking of hydroxyproline-rich glycoproteins (HRGPs) in the plant cell wall is acknowledged to contribute to the formation of a resistant barrier against pathogen infection. We have isolated, from suspension cultured potato (Solanum tuberosum L. cv. Desiree) cells, two forms of soluble HRGP, a cross-linked and a monomeric form; the latter can be converted to the cross-linked form by incubation with tomato extensin peroxidase and H2O2. The monomeric form was purified by Sephacryl S-200 gel-filtration, reverse-phase high performance liquid chromatography and Mono-S cation-exchange chromatography into two isoforms (A, a minor form; B, a major form). The properties of the B isoform were further investigated. A quantitative enzyme-linked immunosorbent assay of the B isoform, using tomato extensin antiserum, showed a titration curve at a high antibody-dilution range comparable to that of purified tomato extension monomer (M.D. Brownleader and P.M. Dey, 1993, Planta 191: 457-469). The amino acid and carbohydrate compositions were similar to those of tomato extensin, but did not match well with the other two HRGPs from potato, potato lectin and potato bacterial agglutinin. These observations demonstrate the similarities of the B isoform to extensin. The homogeneity of the B isoform was demonstrated by its ability to be fully cross-linked in vitro, leaving no residual protein, into a high-molecular-weight form by the action of extensin peroxidase. The trifluoroacetic acid-deglycosylated sample migrated as a single protein band on sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Moreover, SDS-PAGE and matrix-assisted laser desorption/ionisation-time of flight mass spectrometry indicated a molecular weight of approximately 67 kDa. Circular dichroism spectroscopy demonstrated that the molecule possess an extended polyproline II helix conformation with no evidence of alpha-helix or beta- sheet secondary structure. In conclusion, we refer to this HRGP as potato extensin. As proposed for other extensins, potato extensin is likely to play a role in cell wall architecture and plant disease resistance. PMID- 9202493 TI - Flow cytometry, sorting and immunocharacterization with proliferating cell nuclear antigen of cycling and non-cycling cells in synchronized pea root tips. AB - In the 3-d-old 2-mm root tip of Pisum sativum L. cv. Lincoln the percentage of actively proliferating cells is estimated to be 70%. The remaining cells are non cycling and arrested with 2C and 4C DNA content in G0 and in G2Q, respectively. In this work we studied the kinetic significance of these quiescent cells, using the sorting capabilities of flow cytometry and immunofluorescence techniques to detect the proliferation marker PCNA (proliferating cell nuclear antigen) inside cells within the different cell-cycle compartments. While in animal cells, PCNA is present at a high level only in actively proliferating cells, in 3-d-old pea root tips 95% of the cells are PCNA-positive. After flow cytometry and sorting of pea non-cycling nuclear populations, all G2Q nuclei appeared strongly PCNA positive, indicating that these cells had recently left the cell cycle. By contrast, most G0 nuclei showed a low level of PCNA immunofluorescence intensity, as measured by image analysis, with about 25% of the nuclei being PCNA-negative. This small percentage was found to correspond to root cap cells, as could be observed in the root tip section. These are the only cells in the root apical region which are fully differentiated and which, therefore, lack the competence to enter the cell cycle. In contrast, the more or less PCNA-positive G0 nuclei could represent a kinetically heterogeneous population of cells competent to proliferate, but which have either recently left the cell cycle or are progressing to the G0-G1 transition. PMID- 9202494 TI - A pleiotropic Arabidopsis thaliana mutant with inverted root chirality. AB - Circumnutation is an oscillating movement of a growing plant organ that is believed to result from an endogenous rhythmic process intrinsic to growth. Circumnutating organs, as they extend, describe a helical trace. In Arabidopsis thaliana (L.) Heynh. circumnutation is particularly evident in primary roots and occurs, as in most plants, in a right-handed direction when viewed from above in the direction of the growing tips. We have discovered a pleiotropic mutant of Arabidopsis with left-handed root circumnutation. Major abnormalities of the mutant are: (i) a reduced size of all organs, mainly due to a defect in cell elongation or expansion; (ii) a zigzagging pattern of stem pith cells, reminiscent of the "erectoides" phenotype of the lk mutant of Pisum; (iii) roots of the mutant are gravitropic but as they grow, they form tight, left-handed coils. Genetically, the mutant depends on the presence of two independent monogenic recessive factors acting additively. The mutant alleles of both factors alter the growth of the aerial organs in a similar manner but differ at the root level: one mainly produces non-circumnutating roots, the other changes the direction of circumnutation from right to left hand. PMID- 9202498 TI - Race, ethnicity, and sexual health. PMID- 9202499 TI - Disillusioned doctors. PMID- 9202500 TI - Supporting diversity in primary care. PMID- 9202501 TI - Fighting malaria. PMID- 9202502 TI - Commentary: income inequality summarises the health burden of individual relative deprivation. PMID- 9202503 TI - Does it matter who requests necropsies? Prospective study of effect of clinical audit on rate of requests. PMID- 9202504 TI - General practitioner centred scheme for treatment of opiate dependent drug injectors in Glasgow. PMID- 9202505 TI - Recent advances. Diagnosis and treatment of early breast cancer. PMID- 9202506 TI - ABC of mental health. Mental health emergencies. PMID- 9202507 TI - Sexual health--a Health of the Nation failure. PMID- 9202508 TI - HIV and AIDS, other sexually transmitted diseases, and tuberculosis in ethnic minorities in United Kingdom: is surveillance serving its purpose? AB - Experience of disease differs across ethnic groups, and ethnicity is a relevant personal characteristic for descriptive epidemiology. Information about ethnicity and country of birth is omitted from the routine notification of many diseases. HIV infection and AIDS, other sexually transmitted diseases, and tuberculosis have different incidence rates in different ethnic groups in the United Kingdom. Omission of ethnic data from surveillance activities allows such differences in incidence to go undetected and unaddressed. Surveillance data that included ethnic details could guide interventions to reduce inequalities in health between different subpopulations. PMID- 9202509 TI - Is research into ethnicity and health racist, unsound, or important science? AB - Much historical research on race, intelligence, and health was racist, unethical, and ineffective. The concepts of race and ethnicity are difficult to define but continue to be applied to the study of the health of immigrant and ethnic minority groups in the hope of advancing understanding of causes of disease. While a morass of associations has been generated, race and ethnicity in health research have seldom given fundamental new understanding of disease. Most such research is "black box epidemiology." Researchers have not overcome the many conceptual and technical problems of research into ethnicity and health. By emphasising the negative aspects of the health of ethnic minority groups, research may have damaged their social standing and deflected attention from their health priorities. Unless researchers recognise the difficulties with research into ethnicity and health and correct its weaknesses, 20th century research in this subject may suffer the same ignominious fate as that of race science in the 19th century. PMID- 9202510 TI - Save our service. PMID- 9202511 TI - Iron deficiency anaemia. Gastrointestinal endoscopy should always be used. PMID- 9202512 TI - Iron deficiency anaemia. Limit investigations if iron deficiency has no obvious cause. PMID- 9202513 TI - Iron deficiency anaemia. Patients must be screened for coeliac disease. PMID- 9202514 TI - Iron deficiency anaemia. Is important in young children. PMID- 9202515 TI - More teenagers seem to be seeking contraceptive advice from their general practitioner. PMID- 9202516 TI - Increased parity and risk of trisomy 21. Study measured prevalence of Down's syndrome at birth, not incidence. PMID- 9202517 TI - Brain damage in divers. The risk has been underestimated. PMID- 9202518 TI - Brain damage in divers. Cross sectional studies are inconclusive, longitudinal studies are more appropriate. PMID- 9202519 TI - Consent procedure for coronary angioplasty is haphazard. PMID- 9202520 TI - Informed participation in screening is essential. PMID- 9202521 TI - Wait for further evidence confirming effect of interferon beta in multiple sclerosis. PMID- 9202522 TI - Deaths related to methadone have doubled in Lothian. PMID- 9202523 TI - Increased house dust mite allergen in synthetic pillows may explain increased wheezing. PMID- 9202524 TI - Thyroxine should be tried in clinically hypothyroid but biochemically euthyroid patients. PMID- 9202525 TI - Large pragmatic randomised studies of new antiepileptic drugs are needed. PMID- 9202526 TI - Patients from coronary care units should be given their electrocardiograms and carry them at all times. PMID- 9202527 TI - New Zealand priority criteria project. More use should be made of patient oriented quality of life measures. PMID- 9202528 TI - Location, location, location. AB - The essence of parathyroid surgery is finding the diseased gland or glands. Even the most experienced parathyroid surgeons have a finite, albeit small, miss rate. The information above shows that there has been, over the years, a significant amount of effort expended to find a way to localize the glands in reliable fashion. Although current parathormone assays are virtually certain to identity the disease, available localization studies still miss a sizable number of lesions, and the statement, "The best way of localizing the parathyroid glands is to localize an experienced parathyroid surgeon," probably remains valid. Does such a surgeon need an ultrasound or sestamibi scan preoperatively? Probably not. I would note, however, that many such surgeons have written of their experience with these localization studies, suggesting that they usually operate with this information available to them. In the managed care era, with many of these procedures being done by less frequent parathyroid surgeons despite the failure of currently available studies to show any advantage in operative time or success, the high positive predictive value of the sestamibi scan in particular can, I think, be very helpful in focusing the procedure. Certainly, the patient should not be denied this advantage by cost issues, which in this uncommon disease only serve to benefit the balance sheet of the managed care organization. PMID- 9202529 TI - Nonfunctioning islet cell tumors of the pancreas: a difficult diagnosis but one worth the effort. AB - Islet cell tumors of the pancreas usually secrete gastroenteropancreatic peptides causing well-recognized clinical syndromes. Description of these syndromes and the identification of the responsible hormones by radioimmunoassay has led to a better understanding of neuroendocrine regulatory function. More recently, similar tumors have been seen that contain various peptides on immunohistochemical stain but do not secrete these substances sufficiently to cause clinical symptoms. Nonetheless, they have the same malignancy and metastatic rate as most of the functional tumors. Between 1972 and 1996, 44 patients with islet cell tumors have been treated at the Indiana University Medical Center Hospital, and of these 14 have been nonfunctional. Preoperative imaging studies, such as CT scan and endoscopic ultrasound, were able to visualize a lesion but not to make the specific diagnosis, even with fine-needle aspiration. Pancreatic ductal preservation on endoscopic retrograde cholangiopancreatography with CT evidence of a mass should arouse suspicion of an islet cell tumor. Once discovered, all but 1 of the 14 patients has under gone resective therapy, with only 1 postoperative death. Treatment has been aggressive, with 11 of the 13 resected patients undergoing pancreaticoduodenectomy, and 2 others distal pancreatectomy. Four of the seven patients with positive lymph node metastases are dead, while all patients with negative nodes are still alive. Thus far, 10 of the original 14 patients are alive, surviving an average of 32.7 months, with a median survival of 31.1 months. Because these tumors have a better overall prognosis, vigorous attempts at total or subtotal resection should be carried out, since the long-term survival is enhanced by tumor bulk reduction or curative resection when possible. PMID- 9202530 TI - Adjuvant radiation therapy in resectable rectal cancer: should local recurrence rates affect the decision? AB - Adjuvant external beam pelvic radiotherapy (XRT) for resectable rectal cancer has been mandated by the National Cancer Institute because of reported 20 to 50 per cent reductions in local recurrence rates. However, these series' reported local recurrence rates are 18 to 39 per cent in the nonradiated patients, which seems extraordinarily high compared to the 3 to 5 per cent rates reported by surgeons advocating proctectomy with complete mesorectal excision. This fact, coupled with the high cost of XRT ($11,000-$14,000), the risk of radiation injury to small bowel and the neo-rectum, and the failure of XRT to provide any survival advantage, raises questions as to the precise role of XRT for rectal cancer. The purpose of this study was to perform a review of 212 consecutive patients undergoing curative resection via low anterior resection (LAR) or abdominoperineal resection (APR) for rectal cancer between 1989 and 1993, focusing on local and distant recurrence rates and survival. The choice of surgery alone (SUR), preoperative radiation (PRE) (45 Gy), or postoperative radiation (POST) (45-50 Gy) was at the surgeon's discretion. There were no significant differences in male:female ratio (SUR, 83:60; PRE, 14:8; POST, 34:13) or type of procedure (SUR-LAR, 112:APR, 31; PRE-LAR, 5:APR, 17; POST-LAR, 30:APR, 17) between the groups. There were no significant differences in age between the preoperative and postoperative radiation groups (PRE, 64.0 +/- 2.4; POST, 59.2 +/ 1.7); however, age was significantly different (P < 0.05) between the surgery alone and the postoperative radiation groups (SUR, 68.5 +/- 0.8; POST, 59.2 +/- 1.7). With a median follow-up of 49 months, there were no significant differences in local recurrence (SUR, 4.2%; PRE, 4.5%; POST, 2.1%); however, there was a significantly longer survival for the SUR group compared to the other groups (SUR, 45.9 months; PRE, 36.4 months; POST, 39.3 months; P < 0.05 least significant difference). The PRE group also had shorter survival compared to the other groups when only Stage II and III lesions were studied (S, 40.0 months; PRE, 28.3 months; POST, 39.3 months). Local recurrences based on TNM stage were: T1N0 (S, 0 of 27; PRE, 0 of 3); T2N0 (S, 4 of 4S; PRE, 0 of 7); T2N1 (S, 0 of 9; POST, 1 of 5); T3,4N0 (S, 2 of 37; PRE, 1 of 9; POST, 0 of 10); and T3,4N1,2 (S, 0 of 21; PRE, 0 of 3; POST, 0 of 30). The results of this series support the contention that proctectomy with complete mesorectal excision yields a 4.2 per cent local recurrence rate without the need for adjuvant XRT. In this series, if all the patients had received adjuvant radiation, an additional $2.2 million would have been added to the costs of medical care. Therefore, the potential risks, costs, and benefits of adjuvant pelvic XRT for rectal cancer must be weighed against optimal benchmarks for local recurrence rate for surgery alone. PMID- 9202531 TI - "Same-day" thyroid surgery: an analysis of safety, cost savings, and outcome. AB - Twenty-three-hour observation or "same-day" thyroid surgery was initiated at our institution for patients with nodular thyroid disease in July 1993. A retrospective review of all patients with nodular thyroid disease who underwent same-day thyroid surgery was performed to determine the safety and cost effectiveness of this approach. Eighty consecutive patients with nodular thyroid disease underwent thyroidectomy, followed by < or = 23 hours of observation in 71 (88%) and a planned hospital admission in 9 (12%) patients. Hospital admission preceded thyroidectomy in four patients (5%) because of transient ischemic attacks (one), airway obstruction (one), pulmonary disease (one), and suppurative thyroiditis (one). Five patients (6%) had a planned postoperative admission because of concominant modified neck dissection (two), median sternotomy (two), or soft tissue tumor resection (one). Of the 71 patients who underwent same-day thyroid surgery, 47 had near-total or total thyroidectomy, 20 lobectomy, and 4 completion thyroidectomy. Morbidity consisted of hematoma in one, recurrent laryngeal nerve paresis in two, and transient hypocalcemia in eight patients. Only 1 of the 71 patients required subsequent hospitalization for an anxiety attack. There was no mortality. Twenty-three-hour observation was associated with a 32 per cent and a 47 to 56 per cent reduction in cost for unilateral and bilateral thyroidectomy, respectively. Same-day thyroid surgery is a safe and costeffective approach for patients with nodular thyroid disease. PMID- 9202532 TI - Intraoperative ultrasound in the management of liver neoplasms. AB - The purpose of this study was to determine the impact of intraoperative ultrasound (IOUS) on the management of patients with neoplasms of the liver. Fifty-nine patients with liver neoplasms (primary, 6; metastatic, 53) and without pre- or intraoperative evidence of extrahepatic disease underwent laparotomy for possible liver resection. Preoperative imaging studies included external ultrasound (n = 12), magnetic resonance imaging (n = 11), and/or computed tomography (n = 57). Intraoperative evaluation on all patients included inspection, bimanual palpation, and ultrasonography. External ultrasound, magnetic resonance imaging, and computed tomography identified all intraoperatively confirmed liver neoplasms in 33, 45, and 67 per cent of cases, respectively. Unsuspected neoplasms were identified in 12 patients (20%) by inspection/palpation and in 19 patients (32%) by IOUS. In eight patients (14%), the occult neoplasms were identified only IOUS, and in one patient the neoplasms were identified only by inspection/palpation. Occult neoplasms identified by IOUS were characterized by small size (less than 2 cm). Findings from the intraoperative evaluation, such as unsuspected neoplasms and vascular proximity or invasion, altered the preoperative plan in 20 (34%) patients. Inspection, and particularly palpation, identifies a number of preoperatively unsuspected liver neoplasms. Intraoperative ultrasound, however, is the most sensitive method for detection of liver neoplasms and influences the operative management in a substantial number of patients. PMID- 9202533 TI - Pancreatic trauma: a ten-year multi-institutional experience. AB - Our objective was to determine the incidence, management, and outcome of traumatic pancreatic injury. A retrospective review was performed of all patients with pancreatic injury admitted to two Level I trauma hospitals over a 10-year period. Comparisons were made with Chi square or Fisher's exact tests. Of 16,188 trauma admissions, 72 patients (0.4%) had pancreatic injury. The mean age was 30 years, and 30 patients (69%) were male. Mechanism of injury was gunshot in 32 (45%), blunt in 27 (37%), and stab wound in 13 (18%). The pancreas was involved in 1.1 per cent of patients with penetrating injuries compared to 0.2 per cent with blunt injuries (P < 0.01). There were 18 grade I (25%), 32 grade II (45%), 16 grade III (22%), and 5 grade IV (7%) injuries. Initial diagnosis was made intraoperatively in 63 patients and by computed tomography in 8. The mean injury grade was significantly lower on computed tomography compared to surgical exploration (0.4 vs 2.0; P < 0.05). Operative procedures included distal pancreatectomy in 23 (32%), exploration only in 22 (31%), external drainage in 13 (18%), pancreatorrhaphy in 4, internal drainage in 2, and proximal resection in 2. Mortality was 16.6 per cent and was not related to the mechanism or grade of injury. Mean Injury Severity Score and transfusion requirements were significantly greater in patients who died (P < 0.05). Morbidity occurred in 30 patients (42%), including pancreatic fistula (11%), pancreatitis (7%), and pancreatic pseudocyst (3%). Six patients (8%) developed intra-abdominal abscesses, and all had associated liver or intestinal injuries. In patients with grade I and II injuries, morbidity was higher with external drainage compared to exploration without drainage. Pancreatic injury is infrequent and is more often associated with penetrating trauma. Diagnosis is most commonly made by exploration and cannot be excluded by computed tomography. Drainage of low-grade injuries may not be necessary. Morbidity and mortality in patients with pancreatic trauma is significant and is primarily due to associated injuries. PMID- 9202534 TI - Hepatic resection for metastatic colorectal cancer. AB - One-hundred thirty-one primary hepatic resection for colorectal secondary tumors were performed at Rush-Presbyterian-St. Luke's Medical Center between 1975 and 1993. Perioperative mortality occurred in five patients (3.8%). Twenty-three patients had minor morbidities (18%); major morbidity occurred only in the five patients who died. Curative resections were performed in 107 patients. Overall actuarial survival at 2, 3, and 5 years was 62, 42, and 25 per cent, respectively. Patients with extrahepatic disease (5-year survival, 0% vs 27%; P = 0.049) and positive resection margins (0% vs 30%; P < 0.001) had significantly poorer survival. Among the curative resections, patients who had metachronous hepatic resections did significantly better than those who underwent synchronous colon and hepatic resections (35% vs 13%; P = 0.002). This survival benefit persisted when comparison was restricted to patients with synchronous metastases. Age, sex, race, number of lesions, site of colon primary resection, blood transfusion, disease-free interval, and extent of resection had no effect on survival. All patients who are acceptable surgical risks with potentially resectable metastatic colorectal cancer confined to the liver should undergo exploration. Assessment of resectability should include intraoperative ultrasound in all patients to maximize the probability of tumor clearance. PMID- 9202535 TI - Cholescintigraphy in the diagnosis of bile leak after laparoscopic cholecystectomy. AB - Bile leaks are a recognized complication of laparoscopic cholecystectomy (LC). Different diagnostic approaches have been employed when this condition is suspected. We present our experience with cholescintigraphy as a primary imaging technique for the detection of bile leaks. The medical records of all patients who had cholescintigraphy after LC during a 58-month period were reviewed. Patients were selected for cholescintigraphy if fever unusual abdominal pain, nausea, vomiting, or jaundice were present beyond 36 hours after LC. Bile leaks were suspected in 25 out of 744 patients (3.36%). The nuclear imaging study was true positive in 7 cases and true negative in 18 cases, for a 100 per cent sensitivity, specificity, and accuracy in the detection of bile leaks. Five patients were treated by endoscopic retrograde cholangiopancreatography with stent and/or sphincterotomy, and two patients underwent exploratory laparotomy. None of the patients who underwent endoscopic retrograde cholangiopancreatography required peritoneal drainage. We conclude that cholescintigraphy is sensitive and accurate in the diagnosis of bile leaks. Its use along with a high index of suspicion of a bile leak may prevent the development of bile peritonitis. PMID- 9202536 TI - Herpes simplex thymidine kinase gene transfer is required for complete regression of murine colon adenocarcinoma. AB - The herpes simplex thymidine kinase (HStk) gene induces regression of epithelial tumors after ganciclovir (GCV) administration. This observation has been attributed to both gene transfer and metabolic cooperation between cells (the bystander effect). This study evaluates the relative roles of the bystander effect and gene delivery of the HStk gene by the LTKOSN.2 vector. MC38 colon adenocarcinoma cells, syngeneic for C57/B16 mice, were used. Whereas in vitro proliferation assays demonstrated a bystander effect, significantly greater inhibition of proliferation occurred with HStk gene transfer. In mixtures containing 75 per cent MC38 cells with no vector (MC38 NV) and 25 per cent MC38 pretransduced with LTKOSN.2 (MC38 TK), proliferation was inhibited by 62 +/- 5 per cent. In mixtures containing 75 per cent MC38 NV with 25 per cent HStk vector producing cells (LTKOSN.2 VPC), proliferation was inhibited by 97 +/- 1 per cent. In vivo subcutaneous mixture experiments utilized MC38 NV cells inoculated at a 1:1 ratio with various treatment cell groups followed by administration of GCV. Tumor volumes (mean +/- standard error) at 30 days were: 264 +/- 66 mm3 for MC38 TK, 0 for LTKOSN.2 VPC, 1009 +/- 335 mm3 for lacZ VPC (beta-galactosidase VPC), and 1012 +/- 212 mm3 for NIH3T3 (nontransduced cells). These data suggest that in vivo, the bystander effect alone causes tumor inhibition, but gene transfer is necessary for complete tumor elimination in immunocompetent mice. PMID- 9202537 TI - T1a and T1b breast cancer: a twelve-year experience. AB - To understand the prevalence of axillary node metastasis and survival of patients with T1a and T1b breast cancers, we reviewed the experience at a large community hospital. All patients in the William Beaumont Hospital tumor registry with breast cancer treated between January 1983 and November 1995 were evaluated for tumor size, age, cell type, and the presence or absence of axillary node disease. Long-term survival was evaluated in patients treated between 1983 and 1992. The patients were defined as premenopausal or postmenopausal based on age (49 years or less, premenopausal; 50 years or greater, postmenopausal). Of the 4590 patients treated for breast cancer from 1983 to 1995, 915 had tumors 1.0 cm or less in size. Of 181 patients who had T1a cancer, 27 were premenopausal, and 154 were postmenopausal. Twenty-three premenopausal patients had axillary lymph nodes examined, two (8.7%) had histologically positive lymph nodes. Of 118 postmenopausal patients who had axillary nodes examined, six (5.1%) had positive lymph nodes. In those with T1b tumors, 130 patients were premenopausal; 604 patients were postmenopausal. Of these, 119 premenopausal patients had axillary nodes examined, and 29 (24.4%) had positive lymph nodes. Of 464 postmenopausal patients who had axillary nodes examined, 66 (14.2%) had positive nodes. The overall, disease-free, and tumor-specific survival rates for patients with T1a tumors were 93.8, 87.5, and 93.8 per cent (premenopausal) and 86.2, 95.4, and 95.4 per cent (postmenopausal), respectively. These survival rates for patients with T1b tumors were 87.8, 87.8, and 91.1 per cent (premenopausal) and 82.9, 88.5, and 92.9 per cent (postmenopausal), respectively. Premenopausal T1b patients had a higher rate of nodal involvement than postmenopausal T1b patients (P = 0.011). Postmenopausal T1b patients had a higher nodal metastasis rate than postmenopausal T1a patients (P = 0.01). T1b patients had a higher rate of axillary involvement than did T1a patients (P = 0.0018). Based on the rate of axillary lymph node metastasis and survival statistics, there may be a role for axillary node dissection in select patients with tumors less than 1.0 cm. in size. PMID- 9202538 TI - Long-term outcome after ileocecal resection for Crohn's disease. AB - The decision to operate on ileocecal Crohn's disease is usually tempered by concern for early recurrence and the potential for multiple small bowel resections that will render the patients a gastroenterological cripple. However, delays in surgical management may unnecessarily prolong the patient's disease state and risk complications from both medications and unchecked disease. The aim of this study was to report the long-term clinical outcome of patients undergoing ileocecal resection for Crohn's disease between 1970 and 1993. One hundred eighty one patients underwent ileocecal resection for Crohn's disease during the study period, with a median follow-up of 14.3 years. The mean age at the first resection was 32.7 +/- 0.9 years, and the male female ratio was 79:102. The indications for the initial resection were intractability in 119 (68.4%), obstruction in 45 (25.9%), enteric fistula in 27 (15.5%), perforation in 16 (9.2%), intra-abdominal abscess in 7 (4.0%), and hemorrhage in 5 (2.9%). Postoperative complications included prolonged ileus in 13 (7.5%), pneumonia/atelectasis in 15 (8.6%), wound infection in 11 (6.3%), urinary tract infection in 10 (5.7%), intra-abdominal abscess in 7 (4.0%), and wound dehiscence in 1 (0.6%). There were no operative mortalities. Fifty-six (30.9%) developed a recurrence requiring further surgery, with the mean time interval between initial ileocecal resection and operation for recurrence being 72.3 +/- 7.6 months. A second recurrence developed in 19 patients (10.5%) with a mean time interval of 52.3 +/- 8.3 months. The most frequent sites of first recurrence were the preanastomotic ileum in 49 (87.3%), the postanastomotic colon in 10 (17.9%), other colonic sites in 16 (28.6%), and other small bowel sites in 2 (3.6%) and other sites in 4 (7.1%). The types of resection for first recurrence were ileal resection in 28 (50%), right hemicolectomy in 17 (30.4%), segmental colectomy in 6 (10.7%), total proctocolectomy in 3 (5.4%), and proximal small bowel resection in 2 (3.6%). The long-term follow-up of this patient cohort indicated that 125 (69.1%) had only one resection, 37 (20.4%) required two resections, 15 (8.3%) required three resections, 4 (2.2%) required four resections. The results indicate that ileocecal resection of Crohn's disease had a high rate of disease control obtained with low morbidity, and a low frequency of three or more bowel resections (2.2%). Therefore, surgical resection of ileocecal Crohn's disease should not be unduly delayed for fear of risking short bowel syndrome. This approach should minimize overall disease-related patient morbidity by avoiding long periods of chronic illness. PMID- 9202539 TI - Multimodality staging optimizes resectability in patients with pancreatic and ampullary cancer. AB - Few patients with pancreatic cancer have resectable disease at the time of diagnosis, and a variety of nonsurgical techniques are available to provide effective palliation of jaundice and pain. Accurate preoperative staging is essential to identify patients with unresectable disease, thereby minimizing unnecessary surgery. Currently used diagnostic tests include contrast-enhanced computerized tomography (CT), visceral angiography, endoscopic ultrasound, and laparoscopy, but their utility remains controversial. To evaluate the accuracy of these various diagnostic tests, 30 consecutive patients with histologically proven pancreatic or ampullary adenocarcinoma treated between 1992 and 1996 were evaluated. All 30 patients had contrast-enhanced CT and laparoscopy, 22 patients (73%) had visceral angiography, and 16 patients (53%) had endoscopic ultrasound. Individual and combined predictive values of resectability and unresectability as well as the sensitivities and specificities were determined for all diagnostic tests and compared with intraoperative findings. When CT, visceral angiography, and laparoscopy were combined, the predictive values of resectability and unresectability were 75 and 90 per cent, respectively, with a sensitivity of 75 per cent and a specificity of 90 per cent. Therefore, the combined use of selected diagnostic tests proved more effective than any single diagnostic test for accurately staging patients with pancreatic head and ampullary cancers and should be considered to minimize unnecessary surgery. PMID- 9202540 TI - Tubular carcinoma of the breast: an institutional review. AB - Tubular carcinoma of the breast is a rare, but distinct, well-differentiated histologic subtype of invasive carcinoma, known for its favorable prognosis. Review of the literature reveals controversy relative to the frequency of tubular carcinoma, the mammographic appearance, the incidence of lymph node metastases, and the need for axillary node dissection. In consideration of these variables and because of the concern that this type of breast cancer was being surgically over-treated, this review was undertaken. Through the use of our tumor registry, a retrospective review of patients with invasive breast cancer was carried out from January 1984 to September 1995. Of 1483 total cases of invasive breast carcinoma, 22 (1.5%) had a diagnosis of pure tubular carcinoma (> 90% tubular component). The mean age was 58 years (range, 37-80). In 14 patients, the lesion was detected solely by mammography with a mean size of 1.0 cm (range, 0.5-1.5 cm). The mean pathologic tumor size was 1.2 cm (range, 0.5-2.9 cm). The mean number of nodes, in 22 axillary specimens, was 19 (range, 8-30). In one patient, there was lymph node metastasis to a single node (4.5% incidence), which demonstrated tubular characteristics. Presently, 18 of the patients are alive and disease free, with a mean follow-up of 3.5 years (range, 4 months to 9 years). Our study confirms the low incidence of pure tubular carcinoma, 1.5 per cent, with the lesions being small, 1.2 cm in mean size. The mammographic lesions had no unique features that would distinguish tubular from other invasive carcinomas. With the small tumor size and the overall excellent prognosis, these patients are ideal candidates for breast preservation. Most importantly, the review did demonstrate that even in pure tubular breast carcinoma, lymph node metastases, though rare, can occur. PMID- 9202541 TI - Percutaneous tracheostomy: a cost-effective alternative to standard open tracheostomy. AB - Percutaneous tracheostomy was initiated as an alternative to open tracheostomy at our institution in December 1993. To assess safety, operative time, and cost, a comparative analysis of percutaneous and open tracheostomies was performed. A retrospective evaluation of all patients who underwent percutaneous tracheostomy (P) from December 1993 to March 1996 was completed. Patients were evaluated for indications for tracheostomy, length of operation, morbidity, and cost. The results were compared with patients who underwent open tracheostomy (O) during the 12 months prior to introduction of the percutaneous technique. Tracheostomy was performed percutaneously in 74 patients and by a standard open technique in 109 patients. Indications for tracheostomy included: chronic ventilator dependence (P, 49 vs O, 58); airway protection (P, 19 vs O, 42); laryngeal dysfunction (P, 2 vs O, 7); and facial trauma (P 6 vs O, 2). The length of operation was 21 +/- 6 minutes and 46 +/- 21 minutes for percutaneous and open tracheostomy, respectively (P < 0.05). Perioperative morbidity occurred in 2 patients (3%) following percutaneous tracheostomy compared to 10 patients (9%) following open tracheostomy (P > 0.05). The mean operating room costs per patient were $1093 and $1370 for percutaneous and open tracheostomy, respectively. Percutaneous tracheostomy is a safe procedure that can be performed in less time and at a lower cost than standard open tracheostomy. PMID- 9202542 TI - Enterostomal complications: are emergently created enterostomas at greater risk? AB - It is not unusual for surgeons to have to construct a enterostoma during an emergency abdominal operation. The enterostomal complications, often overlooked, can be serious for the patient. There are many factors relating to stoma complications. The purpose of this paper is to determine whether the emergency status of an operation is an independent risk. Over a 19-year period from 1976 to 1995, there were 1758 enterostomas constructed at our institution. Fifty-nine per cent were for emergent situations, defined as any operation performed for peritonitis, obstructions, or massive hemorrhage. The data pertaining to complications was compiled by the enterostomal therapist and prospectively recorded into an institutional database. Complications were characterized as skin problems, parastomal problems (infection, separation), retraction, stenosis, necrosis, prolapse, and herniation. There were 624 (35%) patients with recorded complications. It was not uncommon for a patient to have more than one complication. There were 500 (55%) skin problems, 111 (12%) parastomal problems, 104 (11%) retractions, 33 (4%) stenoses, 112 (12%) necroses, 28 (3%) prolapses, and 19 (3%) enterostomas herniated. Overall, there were 1044 emergently created enterostomas, and we found that 356 (34%) patients had a complication. The most common indications for emergency laparotomies were abdominal gunshot wounds (40%), bowel obstruction (20%), bowel perforation other than by gunshot or stab wound (15%), and diverticulitis (8%). Among the nonemergently created enterostomas (714), there were 268 (37%) with complications (P = 0.015). Our findings suggest that emergently created enterostomas are not at greater risk for complications, except for the ileostomy. Although further analysis of this particular subset must be undertaken, the technical intricacies of an ileostomy, including preoperative marking of the site, might have an important role. PMID- 9202543 TI - HIV, hepatitis B, and hepatitis C in the Code One trauma population. AB - The resuscitation of a trauma patient is a very hectic and seemingly chaotic situation. The nature of the situation increases the likelihood that a health care worker will receive an accidental needle stick while caring for the patient. There is also a common belief that there is a higher prevalence of communicable bloodborne diseases in the trauma population, and human immunodeficiency virus (HIV) causes the greatest amount of fear. This belief of higher prevalence communicable diseases in the trauma population is supported by the literature. We sent blood from our Code One (life-threatening) trauma population for analysis for HIV, hepatitis B surface antigen, and hepatitis C antibodies. We found that 0.52% of our trauma population had HIV. We also found that 1.5 per cent had hepatitis B surface antigen, and 13.8 per cent had hepatitis C antibodies. Our prevalence of HIV and hepatitis B is similar to that found in other studies of the trauma populations and higher than the general population. Our hepatitis C prevalence is surprising, considering the very low reported prevalence in the general population. Hepatitis C may be a more significant threat to the health of medical personnel than previously believed. PMID- 9202544 TI - Effectiveness of the thyroid scan in evaluation of the solitary thyroid nodule. AB - Solitary, palpable thyroid nodules are common, but only a small percentage are malignant. It is important to evaluate these nodules in a cost-efficient manner that avoids missing a cancer. Historically, radioisotope imaging has played a major role in the workup of thyroid nodules; however, with the advent of fine needle aspiration biopsy (FNAB), this role has become less clear. From 1974 to 1994, 770 patients with a solitary nodule underwent thyroidectomy. Preoperatively, 471 had thyroid scans, and 149 had FNAB. The incidence of carcinoma in nodules excised on the basis of thyroid scan was 23 per cent, whereas the incidence of carcinoma was 37 per cent when FNAB was used (P = 0.003). Fine needle aspiration was a significantly better predictor of malignancy than thyroid scan and resulted in a smaller proportion of excisions for benign nodules. Thyroid scan provided little additional information in those patients who underwent FNAB. Because thyroid scans add little in determining which nodules require surgical excision, they should no longer be a routine part of the evaluation of a solitary thyroid nodule. PMID- 9202546 TI - On the enlargement of the normal congenitally solitary kidney. AB - OBJECTIVE: To determine the anatomy of the normal congenitally solitary kidney and to complement an earlier investigation of the major functional parameters of congenitally and acquired normal solitary kidneys. MATERIALS AND METHODS: A healthy congenitally solitary kidney was compared anatomically with a normal control kidney from an equal pair in a man of similar height and age. RESULTS: The solitary kidney was about 1.8 times heavier than the control; the diameter of its glomeruli and convoluted tubules were similar to those of the control kidney but there were twice as many glomeruli in the solitary kidney than in the control. The total mass of glomeruli, as a percentage of the renal mass (about 5%), was the same for both kidneys and essentially similar to that for mammals in general. However, the total mass of glomeruli as a percentage of cortical mass was considerably greater in the congenitally solitary kidney than in the control, suggesting a relative deficit of convoluted tubular mass in the solitary kidney. CONCLUSION: The normal congenitally solitary kidney is hyperplastic, i.e. there are more elements (nephrons), and apparently not hypertrophic, i.e. having larger nephrons. PMID- 9202545 TI - Experience and current status of research into the pathophysiology of nocturnal enuresis. PMID- 9202547 TI - Ischaemia-reperfusion injury in the rat kidney: the effect of preconditioning. AB - OBJECTIVES: To design and establish a model to examine whether brief periods of renal artery occlusion (ischaemic preconditioning, IP) confers protection from the effects of a subsequent period of ischaemia and reperfusion of the rat kidney. MATERIALS AND METHODS: Ninety rats were randomized into six groups, i.e. sham-operated controls; IP alone; a 20 or 40 min period of left renal ischaemia (RI) alone; and IP followed by a 20 or 40 min period of RI. Preconditioning involved the sequential clamping of the left renal artery for 4 min and its release for 11 min, a total of four times, a 'critical interval' of 30 min before the ischaemic insult. Left renal tissue integrity was determined by dimercapto succinic acid (DMSA) radionuclide imaging on a gamma-camera both immediately (day 0) and 2 and 9 days later. Acute tubular necrosis was also assessed histologically. RESULTS: RI for 20 min resulted in a significant decrease in left renal tissue integrity on day 2 only (P < 0.001), whereas RI for 40 min caused significant left renal dysfunction on day 0, day 2 and day 9 (P < or = 0.01). For a given duration of ischaemia, there was no significant difference between results from (IP + RI) rats compared with RI-only rats at any of the three times. There was no significant alteration in renal tissue integrity in the IP-only rats compared with sham-operated controls. Histological findings paralleled the data obtained from DMSA uptake. CONCLUSIONS: The IP regimen and 30 min 'critical interval' confers no protection to the kidney from a 20 or 40 min ischaemic episode. The IP regimen itself appears to have no effect, confirming the validity of our experimental model. PMID- 9202548 TI - Use of patient-controlled analgesia in extracorporeal shockwave lithotripsy. AB - OBJECTIVE: To assess the advantages of patient-controlled analgesia (PCA)-in patients undergoing extracorporeal shockwave lithotripsy (ESWL) for urinary stones. PATIENTS AND METHODS: Between December 1995 and May 1996, a prospective study was carried out on 100 patients who underwent ESWL for urinary stones. The patients were assigned to two equal groups, one receiving PCA and the other pethidine (control). Patients in the PCA group self-administered varying doses of intravenous alfentanil according to their pain tolerance, while those in the control group were given a single bolus dose of 1 mg/kg body weight intravenous pethidine by the attending urologist before the start of the procedure. The stone site, maximum energy level achieved, number of shock waves given, duration of procedure, pain scores, patient tolerance and acceptance were recorded to assess the efficacy of PCA compared with analgesia controlled by the physician. RESULTS: Both groups were matched for age, body weight, stone location and number of shocks given. The PCA group received a mean of 1.03 mg alfentanil while the control group received a mean of 62.5 mg pethidine. The maximum discharge voltage of 16 kV was achieved in all but one patient (98%) in the PCA group whereas only 21 patients (42%) in the control group attained this level. The mean treatment duration was less in the PCA group (32.8 min) than in the control group (44.5 min), the mean pain score lower (3.76 and 4.62, respectively) and the incidence of nausea and vomiting much less (22% and 60%, respectively). In addition, all 21 patients in the PCA group who had received intravenous pethidine during previous sessions of ESWL chose PCA as the better form of analgesia. There were no adverse effects in the PCA group except for one patient whose arterial oxygen saturation decreased transiently. CONCLUSION: PCA enables the urologist to achieve better patient compliance through better pain control; its application has maximized the use of lithotripsy and the patients' acceptability for this form of analgesia is confirmed. We recommend that this form of analgesia be used for ESWL. PMID- 9202549 TI - Endovascular treatment of renal artery aneurysms with conventional non-detachable microcoils and Guglielmi detachable coils. AB - OBJECTIVE: To evaluate the efficacy and safety of endovascular occlusion of true renal artery aneurysms (RAAs) with conventional non-detachable microcoils (NDCs) and Guglielmi detachable coils (GDCs). PATIENTS AND METHODS: Over a 5-year period, 12 RAAs were treated by endovascular selective embolization. Four RAAs were occluded using NDCs and eight were treated with GDCs. All coils were delivered through a microcatheter. Eight RAAs were located in the bifurcation of the main renal artery, two in the main renal artery and two were intrarenal. Before treatment, four patients presented with hypertension, one associated with renal infarction and a second had flank pain due to microembolization. Two other patients had renal infarction, associated with haematuria in one; one other patient also had haematuria and five patients were asymptomatic. All patients were followed using clinical and angiographic examinations after 6 months, 1 and 2 years. RESULTS: All RAAs were occluded successfully. In two patients treated with NDCs there were minor complications, i.e. one subsegmental peripheral infarction and one misembolization, both without clinical symptoms. In the group treated with GDCs there were no complications. Five of seven patients were clinically improved, while two patients remained clinically unchanged. CONCLUSION: Superselective endovascular treatment of RAAs with microcoils is a safe, efficient, and less invasive alternative to surgical treatment. The high flexibility and softness of the GDC and the controlled detachment enables a safer and more complete occlusion of RAA than current alternatives. PMID- 9202550 TI - Effect of sucralfate on induced chemical cystitis in rabbits. AB - OBJECTIVE: To study the histological effect of locally applied sucralfate on the urinary bladder in rabbits with chemically induced cystitis. MATERIALS AND METHODS: Sucralfate (1 g dissolved in 10 mL saline) was instilled into the bladders of six rabbits, while four controls had no instillation. The procedure was repeated 4 days later and followed immediately by 10 mL of 2% formaldehyde infused into the bladder and retained for 10 min in all 10 rabbits. After careful flushing with saline, 10 mL sucralfate solution was again instilled in the study group and 10 mL of saline only in the control group. On day 6, the sucralfate instillation was repeated in the six treated animals. All the rabbits were then killed on day 8-9, their bladders removed and examined macroscopically and histologically. RESULTS: Formaldehyde instillation was quickly followed by gross haematuria in all animals. It subsided in the group treated with sucralfate, but not in the controls. Examination revealed only slight changes in the former group, but severe inflammatory lesions in the latter. CONCLUSIONS: Intravesically instilled sucralfate protected rabbit bladder urothelium against chemically induced cystitis. Clinical trials of intravesical sucralfate seem warranted in patients irradiated for pelvic cancer or with severe cystitis caused by cytotoxic drugs. PMID- 9202551 TI - A case-control study to examine any association between idiopathic detrusor instability and gastrointestinal tract disorder, and between irritable bowel syndrome and urinary tract disorder. AB - OBJECTIVE: To assess whether there are common malfunctions (e.g. of the autonomic nervous system and smooth muscle) that underlie disorders of the urinary and gastrointestinal tracts by determining whether there is an increased prevalence (i) of urinary symptoms in patients with irritable bowel syndrome (IBS) and (ii) of gastrointestinal symptoms in patients with idiopathic detrusor instability (IDI). PATIENTS AND METHODS: Questionnaires were sent to patients with a diagnosis of IBS or IDI who were seen in the Departments of Gynaecology, Gastroenterology and Urology at the John Radcliffe and Churchill Hospitals, Oxford, during the 3 year period 1993-1995. The questionnaires were also distributed to control patients who were recruited from the day-surgery unit of the Churchill Hospital. Of 236 questionnaires sent out, 168 replies were analysed; 64 from patients with IBS, 49 from patients with detrusor instability and 55 from controls. The questionnaire included questions about micturition and defecatory behaviour (frequency, regularity, urgency, continence, pain, and ease in passing urine and stools). RESULTS: Patients with IBS were more likely to experience certain urinary symptoms than controls (nocturia, urgency and some forms of urinary urge incontinence) and patients with IDI were as likely as patients with IBS to experience gastrointestinal symptoms more frequently than controls. Control patients showed an unexpectedly high probability of experiencing many of the gastrointestinal and urinary symptoms. CONCLUSIONS: The frequent occurrence of symptoms in control patients makes the significance of the results less clear, but the association between certain symptoms of urinary tract disorder and patients with IBS, and of symptoms of gastrointestinal tract disorder with patients with IDI, suggests that they may share some common underlying dysfunction. PMID- 9202552 TI - Diagnosis and management of high-pressure peristaltic contractions in cystoplasties. AB - OBJECTIVE: To define the aetiology of and therapeutic strategy for high-pressure peristaltic contractions within colo- and caecocystoplasties associated with symptoms of frequency, urgency and urge incontinence. PATIENTS AND METHODS: In a series of over 150 subtotal supratrigonal colo- and caeco-cystoplasties, nine patients were identified with such symptoms. These patients underwent videocystometrography (VCMG) confirming the presence of phasic peristaltic contractions of > 35 cmH2O. RESULTS: In three cases, there was urodynamic evidence of outlet obstruction and symptoms resolved when the obstruction was surgically relieved. In the other six cases, high-pressure peristaltic contractions were present without bladder outlet obstruction. The symptoms did not respond to anticholinergic medication. Three of the six patients had tolerable symptoms which did not warrant further intervention. The other three cases had disabling urge incontinence and underwent ileal patch cystoplasty. The symptoms resolved in all three cases, although later recurred in one patient, probably due to incomplete division of the taenia coli during ileal patch cystoplasty. CONCLUSION: It appears that colonic smooth muscle can develop high pressure contractions in response to neobladder outlet obstruction. Relieving the obstruction ameliorates symptoms associated with these contractions and reduces the magnitude of the peristaltic waves. Symptoms related to high-pressure peristaltic contractions without neobladder outlet obstruction do not respond to anticholinergic medication but can be successfully treated by ileal patch cystoplasty. PMID- 9202553 TI - Urodynamic investigations in reversed ileal seromuscular enterocystoplasty: an experimental study in a rabbit model. AB - OBJECTIVE: To investigate the efficacy of the reversed ileal seromuscular enterocystoplasty (RISMEC) technique and assess the urodynamic findings. MATERIALS AND METHOD: A large bladder defect was created in 12 New Zealand White rabbits: in six the bladder was closed primarily (control group) and in the remaining six the bladder was augmented using the RISMEC technique combined with omentoplasty. The results were assessed using intravenous pyelography (IVP), voiding cysto-urethrography (VCUG), histopathological investigations and urodynamic studies after 4 and 12 weeks. RESULTS: The IVP and VCUG revealed an apparently normal urinary system and voiding pattern in both groups. Histopathologically, in all rabbits undergoing RISMEC, the serosal surface of the reversed ileum was lined with transitional urinary epithelium; there was no evidence of severe fibrosis, inflammation, stone or mucus-production. Urodynamic studies showed a significant decrease in the mean bladder capacity in the controls, but no significant change in the RISMEC group. The mean bladder compliance decreased from 5.85 to 1.36 mL/cmH2O in the controls (P < 0.02) but there was no significant decrease in compliance in the RISMEC group. CONCLUSION: The results suggest that in this rabbit model, the RISMEC technique combined with omentoplasty increases bladder capacity with minimal graft shrinkage; the peritoneal surface is rapidly covered with transitional epithelium and the integrity of the kidneys preserved. Urodynamic investigations showed neobladders with adequate storage capacity, low intravesical pressures and improved compliance rates. PMID- 9202554 TI - Vesico-ureteric reflux in adult patients with spinal injury. AB - OBJECTIVES: To review the incidence, aetiology, treatment and prognosis of vesico ureteric reflux (VUR) in patients in a regional spinal injuries centre. PATIENTS AND METHODS: A retrospective review of radiological investigations revealed 34 of 447 (8%) patients with VUR on at least one study. The notes of these patients were examined to determine their management and outcome. RESULTS: Most patients developed VUR within 4 years of injury; the underlying intravesical pressure was high in half the patients studied. Patients were managed aggressively with a variety of medical and surgical techniques. The VUR of 15 patients resolved completely and in three patients there was some improvement. Renal function deteriorated in three patients as assessed by isotopic scanning. Two patients were transferred to our unit in end-stage renal failure associated with VUR and died within one year. CONCLUSIONS: VUR continues to be a problem in patients with spinal injury and remains potentially fatal through the effects of high transmitted pressure and infection. Renal failure does not always develop as a consequence of the combination of VUR and high intravesical pressure. No single aetiological factor for VUR was found. With active early treatment, the incidence of VUR can be minimized and long-term complications avoided. PMID- 9202555 TI - Pelvic floor exercises as a treatment for post-micturition dribble. AB - OBJECTIVE: To determine the effectiveness of pelvic floor exercises and urethral milking as treatments for post-micturition dribble. PATIENTS AND METHODS: A method of measuring small amounts of urine loss during normal activity was developed; pads were worn for short periods (< 4 h) and then stored in two sealed plastic bags which were weighed within 72 h. Forty-nine men (age range 36-83 years) drawn from a hospital out-patient population, who had not undergone surgery on the bladder, urethra or prostate gland, entered the study. They were randomly assigned to one of three treatment groups; pelvic muscle exercise, urethral milking or counselling. Participants in each group followed the treatment specific to their group for 12 weeks. At 5, 9 and 13 weeks, urine loss was assessed using the method described. RESULTS: The groups were comparable for age, height, weight and pelvic muscle contraction strength and compliance of the men who completed the study was excellent. The outcome measure (improvement in pad weight gain) was strongly influenced by initial pad weight gain, or degree of urine loss at the start of the study and this was treated as a covariate in an analysis of variance model. After allowing for the effects of initial pad weight gain, the counselling group showed no improvement, the urethral milking group showed an adjusted mean improvement in urine loss of 2.9 g after 13 weeks, compared with 4.7 g in the exercise group. CONCLUSION: Both pelvic floor exercises and urethral milking are effective treatments for post-micturition dribble compared with counselling alone. Pelvic floor exercises were more effective in reducing urine loss than urethral milking in this study. PMID- 9202557 TI - High-energy transurethral microwave thermotherapy for large severely obstructing prostates and the use of biodegradable stents to avoid catheterization after treatment. AB - OBJECTIVE: To assess the use of high-energy transurethral microwave thermotherapy (TUMT) for large severely obstructing benign prostatic hyperplasia (BPH) and to compare the use of a biodegradable stent with that of a urethral Foley catheter after TUMT. PATIENTS AND METHODS: The study comprised 30 men (mean age 71 years, range 49-82) scheduled for prostatectomy for symptomatic BPH. Pre-operative investigations included the measurement of urinary free flow rate, residual urine volume (ultrasonographically), a digital rectal examination, transrectal ultrasonography, a symptom score, cystoscopy, cystometry and pressure-flow. The obstruction was graded according to the Schafer nomogram. The patients were treated using the Prostatron (EDAP-Technorned, France) TUMT system; the software used provided a maximum power of 70 W. Patients were catheterised after treatment with either a Foley catheter or a biodegradable stent. After 3 months, the measurements and obstruction grading were repeated, and the effect of the stent assessed. RESULTS: In the entire group, the mean (SD) free flow increased from 7.7 (2.4) to 14.0 (3.3) mL/s, the residual urine decreased from 125 (86) to 23 (25) mL and the symptom score decreased from 16 (8) to 5 (4). The mean (SD) degree of obstruction decreased from 81.0 (16) to 62.6 (15). The biodegradable stent completely avoided post-treatment retention. CONCLUSION: High-energy TUMT can be used on large severely obstructing prostates with major subjective and objective improvements. The biodegradable stent is useful in relieving the problems of catheterization after treatment. PMID- 9202556 TI - Clinical uroselectivity: evidence from patients treated with slow-release alfuzosin for symptomatic benign prostatic obstruction. AB - OBJECTIVE: To assess the safety profile of slow-release (SR) alfuzosin in the treatment of benign prostatic obstruction (BPO), with special attention to orthostatic blood pressure changes, postural symptoms and efficacy. PATIENTS AND METHODS: Two placebo-controlled studies involving 588 patients (292 receiving SR alfuzosin 5 mg twice daily and 296 a placebo) were pooled; 51% of the patients were > or = 65 years of age and 43% had associated cardiovascular disease including hypertension and/or were receiving concomitant antihypertensive drugs. RESULTS: SR alfuzosin was very well tolerated with an overall incidence of adverse events similar to that of placebo (18.5% and 15.8% of patients, respectively) and an overall incidence of withdrawal from therapy for adverse events lower than that of placebo (3.4% and 5.7%, respectively). Adverse events potentially related to vasodilatation were infrequent with SR alfuzosin (the same incidence as with placebo, i.e. 2.7% of patients) and these adverse events occurred mainly during the first month of alfuzosin treatment. The effect on supine blood pressure was minimal. In the subgroups of elderly and hypertensive patients treated with SR alfuzosin, the cumulative incidence of asymptomatic orthostatic hypotension during the first month of treatment was slightly higher than with placebo with no objective consequences on the incidence of adverse events. The clinical efficacy of SR alfuzosin was confirmed by a significant improvement in urinary symptoms and a significant increase in maximum flow rates. CONCLUSION: SR alfuzosin (10 mg/day) can be administered safely without titration in patients with BPO, even in elderly and hypertensive patients. Its favourable benefit/risk ratio allows alfuzosin to be classified as a clinically uroselective alpha 1-blocker. Specific analysis of orthostatic changes in blood pressure is important when assessing the safety profile of an alpha 1-blocker in patients with BPO. PMID- 9202558 TI - Radiofrequency heat-treatment to the prostate for bladder outlet obstruction associated with benign prostatic hyperplasia: a 4-year outcome study. AB - OBJECTIVES: To evaluate the long-term effect on symptom relief and voiding performance of prostatic radiofrequency thermal treatment for bladder outlet obstruction (BOO) associated with benign prostatic enlargement (BPE). PATIENTS AND METHODS: During 1991-1994, 151 patients (mean age 76 years, range 49-91) with symptomatic BOO associated with BPE (i.e. reduced urinary flow rate and persistent residual urine) but with no clinical evidence of carcinoma of the prostate, were treated using the Thermex II (Direx, Israel) intraurethral radiofrequency (RF) prostatic thermal treatment system. This treatment rates the intraurethral temperature to 48 degrees C for 3 h in a single session under local anaesthesia. The patients were followed using symptom scores and uroflowmetry for 24-48 months after treatment. RESULTS: At 1 year, 70% had a good outcome, but by 3 years this had fallen to 40% and at 4 years to 22%. An increase in mean urinary flow rate, from 9 to 12 mL/s, was sustained throughout the follow-up. There was a reduction in mean residual urine from 83 to 51 mL at 6 months, but this increased to 70 mL by 3 years (not significant). The voiding pressure, which declined at 6 12 months, increased again to near pre-treatment values at 1-2 years. CONCLUSIONS: In patients with symptomatic BPE, prostatic thermal treatment with RF relieves symptoms in 60% for at least 2-3 years. In selected cases it is a useful form of management, as it can be performed as a day-case with minimal risk. Changes in standard objective variables were small but statistically significant for at least one year. The study highlights the importance of waiting for late results from clinical trials. PMID- 9202559 TI - Oxybutynin in the treatment of early detrusor instability after transurethral resection of the prostate. AB - OBJECTIVE: To evaluate the symptomatic and urodynamic effects of oxybutynin in the control of irritative micturitional symptoms during the first week after transurethral resection of benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: Fifty-three patients (median age 67 years, interquartile range 62-72) were included prospectively in a double-blind placebo-controlled study. Pre operatively, uroflowmetry and cystometrography (CMG) were performed, and the post void residual volume (PVR) measured; symptoms were rated according to the Boyarski score. CMG was repeated on the first post-operative day and medication was started on the third day. Before withdrawing the catheter on the fifth day. CMG was repeated. Three days later, symptoms were evaluated according to the Boyarski score and uroflowmetry and the estimate of PVR reassessed. RESULTS: In comparison with placebo, oxybutynin significantly decreased frequency, urgency and detrusor pressure at first sensation of filling. However, oxybutynin did not lower the rate of pre-operative detrusor instability and exerted no effect on the maximal capacity of the bladder and corresponding detrusor pressure. Dryness of mouth was reported in 13% and 65% of patients receiving placebo and oxybutynin, respectively. CONCLUSION: Oxybutynin alleviates early irritative symptoms after transurethral resection of BPH, without consistently modifying bladder urodynamics. PMID- 9202560 TI - The role of free prostate-specific antigen in the diagnosis of prostate cancer. AB - OBJECTIVE: To determine whether the free/total prostate-specific antigen (PSA) ratio can discriminate between patients with prostate cancer or benign prostatic hyperplasia (BPH). PATIENTS AND METHODS: A prospective study was conducted using free and total PSA assays in patients who underwent transrectal-ultrasound guided biopsies indicated by a total serum PSA level of > 4 ng/mL and/or a positive digital rectal examination. Sixty-nine men (median age 68 years, range 57-86) who presented to our out-patient department with symptoms of prostatism were included in the study. Blood samples were drawn from all patients before biopsy. RESULTS: Histopathological examination detected prostate cancer in 17 of 69 (25%) patients and 13 of these 17 patients had a free/total PSA ratio of < 0.15; only 12 of 52 (23%) patients with BPH had a ratio of < 0.15. Receiver operating characteristic analysis indicated a threshold free/total PSA ratio of < or = 0.15 was the optimum discriminatory level. In the whole study group, this threshold had sensitivity, specificity, positive- and negative-predictive values of 76%, 77%, 52% and 91%, respectively. There were 40 patients with serum PSA levels of 4-10 ng/mL and 17.5% (7/40) of these were diagnosed with cancer. Using a free/total PSA ratio of 0.15 would have failed to diagnose two patients of seven with prostate cancer but 30 patients would have avoided a biopsy. In this subgroup, the threshold ratio of 0.15 had sensitivity, specificity, positive- and negative predictive values of 71%, 85%, 50% and 93%, respectively. The rates for a PSA density (PSAD) at a threshold of > or = 0.15 were 71%, 76%, 38%, 93%, respectively. CONCLUSION: These results indicate that using the free/total PSA ratio gives a significant improvement over PSAD and total PSA values alone in the diagnosis of prostate cancer: its use may also enhance the diagnostic accuracy in patients with intermediate PSA levels. PMID- 9202561 TI - Prostatic asymmetry as a risk factor for prostatic carcinoma: serial prostate specific antigen monitoring and cancer detection. AB - OBJECTIVE: To compare the rates of cancer detection in men with a normal, asymmetric, or suspicious prostate on digital rectal examination (DRE) initially and after 3 years of serial monitoring of prostate specific antigen (PSA) level. PATIENTS AND METHODS: Prostatic 'asymmetry' was defined as asymmetric growth of the lateral lobes of the prostate without induration or nodules, as assessed by a DRE. The study included 963 men with no clinical evidence of prostate cancer and whose serum PSA levels were monitored at 4 month intervals. Prostatic biopsy was recommended if the PSA level became persistently abnormal (> 4ng/mL) or increased by > 20% after having been initially abnormal. Cancer detection rates were compared among groups categorized by the initial DRE findings and serum PSA level. RESULTS: On comparing groups with suspicious and normal DREs, and abnormal with normal PSA levels both, as expected, were associated with a statistically significant increase in cancer detection. However, an asymmetric prostate did not carry an increased risk of detecting prostate cancer when compared with a normal prostate, regardless of PSA level. CONCLUSIONS: An asymmetric prostate does not appear to be an independent risk factor for detecting prostate cancer. Therefore, an asymmetric prostate with no abnormality in PSA level should not mandate prostatic biopsy, or even an increase in monitoring frequency above the presently recommended annual interval. PMID- 9202562 TI - A comparison of endorectal magnetic resonance imaging and transrectal ultrasonography in the local staging of prostate cancer with histopathological correlation. AB - OBJECTIVE: To assess the staging accuracy of endorectal magnetic resonance imaging (MRI), using a mid-field system, in patients with clinically localized prostate cancer and to compare the results with transrectal ultrasonography (TRUS). PATIENTS AND METHODS: Twenty patients with clinically localized prostate cancer were prospectively staged with TRUS and endorectal MRI using a 0.5 T magnet. All patients subsequently underwent radical prostatectomy and the results of pre-operative staging were compared with the histological findings. RESULTS: The sensitivity and specificity for diagnosing capsular penetration were 38% and 100%, respectively, for endorectal MRI, and 23% and 86% for TRUS. The sensitivity and specificity for diagnosing seminal vesicle invasion were 100% and 94%, respectively, for endorectal MRI, and 33% and 100% for TRUS. The overall staging accuracy for endorectal MRI was 75% compared with 50% for TRUS. CONCLUSION: Compared with TRUS, endorectal MRI with a 0.5 T magnet provided greater sensitivity and specificity for capsular penetration and increased sensitivity for seminal vesicle invasion. PMID- 9202563 TI - Anaemia associated with androgen deprivation in patients with prostate cancer receiving combined hormone blockade. AB - OBJECTIVE: To describe the incidence, time to onset and extent of anaemia occurring in patients with prostate cancer receiving combined hormone blockade (CHB) and the timing and extent of recovery from anaemia in those patients where CHB was discontinued. PATIENTS AND METHODS: Patients with prostate cancer were evaluated prospectively by physical examination and laboratory tests at baseline and at routine intervals while receiving CHB. Of 142 patients who received CHB, 133 were evaluable for the assessment of anaemia; CHB was discontinued in 76 patients, of whom 64 were assessable for recovery from their anaemia. RESULTS: Haemoglobin levels declined significantly in all patients from a mean baseline of 149 g/L to means of 139 g/L, 132 g/L and 131 g/L at 1, 2 and 3 months, respectively. Haemoglobin levels continued to decline during CHB to a mean nadir of 123 g/L at a mean of 5.6 months of CHB, representing a mean absolute haemoglobin decline at nadir of 25.4 g/L. In 120 of the 133 (90%) patients, the relative decline in haemoglobin at nadir was > or = 10% and was > or = 25% in 17 (13%) others, representing a mean absolute haemoglobin decline in this subset of 42.7 g/L. Significant symptoms related to anaemia occurred in 17 patients (13%). Anaemia and symptoms in these patients were easily corrected with the subcutaneous administration of recombinant human erythropoietin. CONCLUSIONS: The anaemia associated with androgen deprivation is significant and occurs routinely in men receiving CHB. It is normochromic, normocytic, temporally-related to the initiation of androgen blockade and usually resolves after CHB is discontinued. We suggest that patients receiving CHB undergo haematological testing at baseline, 1-2 months after initiating CHB and periodically thereafter. Patients developing anaemia should be questioned about symptoms reflecting physiological compromise (e.g. angina, dyspnoea on exertion). In the absence of other causes, CHB should be suspected in the development of anaemia in patients receiving this treatment. PMID- 9202564 TI - Evaluation by magnetic resonance imaging of the inferior vena cava in patients with non-seminomatous germ cell tumours of the testis metastatic to the retroperitoneum. AB - OBJECTIVE: To assess the role of magnetic resonance imaging (MRI) and magnetic resonance angiography (MRA) in evaluating suspected occlusion of the inferior vena cava (i.v.c.) in patients with abdominal nodal metastases from non seminomatous germ cell tumours, thus giving information that may be helpful in planning surgery and for determining the need for anticoagulant therapy. PATIENTS AND METHODS: Five patients with abdominal nodal metastases in whom occlusion of the i.v.c. was suspected on computed tomography (CT) were imaged using a 1.5 T MRI scanner. The MRI findings were compared with those from CT. RESULTS: The MR images successfully and clearly detected partial and total occlusions of the i.v.c. by both intraluminal thrombus and extrinsic compression. The technique also clearly detected extensive collateral venous circulation in several cases and in one a cavernous transformation of the i.v.c. In all patients, the MRI studies provided better information than that from CT. CONCLUSION: MRI, and particularly MRA, is a comparatively new technique that is non-invasive and offers the potential of evaluating vascular structures with no need for ionizing radiation or contrast media. This technique, if available, should be chosen for imaging the i.v.c. in patients suspected of having compression or occlusion of the i.v.c. PMID- 9202565 TI - Intracavernosal drug-induced erection therapy versus external vacuum devices in the treatment of erectile dysfunction. AB - OBJECTIVES: To determine if there is a significant difference between intracavernosal self-injection and external vacuum devices when compared directly for satisfaction, effectiveness and side-effects. PATIENTS AND METHODS: Fifty men were randomised into two groups and received either instruction on the use of the Osbon ErecAid system or self-injection therapy. After 15 uses, each group completed a questionnaire detailing efficacy, satisfaction and side-effects, and then changed to the other modality after appropriate instruction. A questionnaire was completed by study participants and their sexual partners after using both methods. Patients were followed for 18-24 months using telephone interviews. RESULTS: Forty-four patients (mean age 62.3 years, range 38-84) completed the study. Patients and their partners reported a superior quality of erections with the injection method but the difference did not reach statistical significance. The ability to attain orgasm and the overall satisfaction of the patient and partner with the sexual experience was significantly better when using injections. Side-effects were similar between the modalities. Subgroups analysed for age, duration and aetiology of impotence showed that younger patients (< 60 years), those with a shorter duration of impotence (< 12 months) and those impotent secondary to radical prostatectomy strongly favoured injection therapy (P < 0.05). Overall, of the 44 couples, the final preferences of the patients were 25 (57%), 12 (27%), six (14%) and one (2%) for the injection, vacuum device, both or neither, respectively, and of the partners were 22 (50%), 12 (27%), six (14%) and four (9%), respectively. At 18-24 months, 80% of patients were still using either the vacuum device, injections, or both. CONCLUSION: Both the vacuum device and injections are effective treatment modalities for impotence and are associated with good long-term success. Overall, there was a trend favouring injection therapy over the vacuum device which was most significant in younger patients, those with a shorter duration of impotence, and those impotent secondary to radical prostatectomy. PMID- 9202566 TI - Effects of sildenafil, a type-5 cGMP phosphodiesterase inhibitor, and papaverine on cyclic GMP and cyclic AMP levels in the rabbit corpus cavernosum in vitro. AB - OBJECTIVE: To investigate further the mechanisms of action of sildenafil, a highly selective and potent inhibitor of type 5 cGMP phosphodiesterase (PDE5) that has proved effective in the treatment of erectile dysfunction, by assessing its effect on the in vitro formation of cGMP and cAMP in the corpus cavernosum of the rabbit. MATERIALS AND METHODS: Male New Zealand White rabbits (2.5 kg) were killed and their penises rapidly excised, cut into segments and pooled. Penile segments were then incubated with various concentrations of sildenafil or papaverine. The formation of cGMP was stimulated with increasing concentrations of sodium nitroprusside (SNP) and the cGMP and cAMP concentrations measured by radioimmunoassay. Responses were compared to those obtained with papaverine, which is used therapeutically as an erectogen. RESULTS: In the presence or absence of SNP, sildenafil increased cGMP concentrations in rabbit penile tissue with increasing dose; the increase was greatest (about 28-fold) when cGMP was stimulated with SNP (up to 10 mumol/L). At all stimulatory concentrations of SNP, the effective concentrations for 50% stimulation (EC50) of sildenafil were 430 520 nmol/L. Concentrations of cAMP were unaltered by sildenafil. Papaverine enhanced cGMP formation in response to SNP, but at much higher concentrations than did sildenafil (> or = 10 mumol/L). CONCLUSIONS: Sildenafil specifically increases cGMP levels in rabbit corpora cavernosa; the increase is greater in the presence of SNP indicating that, in vivo, sildenafil may enhance erection by the augmentation of nitric oxide-mediated relaxation pathways. The erectogenic effect of sildenafil is mediated by a specific enhancement of cGMP accumulation in the corpus cavernosum, consistent with the known activity of sildenafil as a potent and highly selective inhibitor of cGMP-specific PDE. PMID- 9202568 TI - The use of the detrusorrhaphy for vesico-ureteric reflux: the way forward? AB - OBJECTIVE: To assess the outcome of all detrusorrhaphies carried out for simple single-system vesico-ureteric reflux (VUR) in children at one institution over a 4-year period. PATIENTS AND METHODS: Twenty-nine patients (11 boys, 18 girls, 43 renal units; mean age at presentation 23 months; range, antenatal to 72 months) were analysed in two groups. Group 1 comprised those undergoing asynchronous bilateral procedures (two patients, four units) and unilateral procedures (15 patients, 15 units), and group 2, those undergoing synchronous bilateral procedures (12 patients, 24 units). The mean (range) follow-up was 17 months (6 39) and 15 months (7-24), respectively. RESULTS: Three patients had antenatal hydronephrosis related to VUR post-natally and 26 had urinary tract infections (13 recurrent, one with haematuria, seven 'breakthrough', one with calculi and four with enuresis). The mean age at operation was 54 months (range 14-167). The mean (range) duration of anaesthesia was 69 min (40-120) in group 1 and 80 min (65-120) in group 2. All patients were catheterized urethrally for a mean (range) duration of 3 days (2-4) in group 1 and 5 days (2-15) in group 2 and the mean hospital stay was 3 days (2-6) and 6 days (4-16), respectively. Post-operative complications occurred in 14 patients including one bladder spasm, five urinary tract infections, two with urinary retention, three with haematuria and one each of pneumonia, epididymo-orchitis, anuria, failure to stent and conversion to Cohen reimplantation. Operative success and clinical success were similar within groups but differed between groups, although not significantly (15 of 17 in group 1 and eight of 12 in group 2). CONCLUSION: Unilateral but not synchronous bilateral detrusorrhaphy seems an appropriate surgical treatment for VUR. PMID- 9202567 TI - Effects of castration on adrenergic, cholinergic and nonadrenergic, noncholinergic responses of isolated corpus cavernosum from rabbit. AB - OBJECTIVE: To investigate the effects of castration and testosterone on the constricting effect of phenylephrine and endothelium-dependent and -independent relaxing effects of different agonists in the corpus cavernosum of male rabbits. MATERIALS AND METHODS: Twenty rabbits were castrated and 10 received testosterone replacement for 1 month after castration; 10 further rabbits underwent a sham operation and acted as controls. One month after operation the rabbits were killed and their penises excised. Strips of corpus cavernosum were used for isometric tension measurements in organ chambers; concentration-response relationships for phenylephrine, carbachol, adenosine and sodium nitroprusside were obtained by adding the reagent cumulatively to the bath. RESULTS: The phenylephrine-induced contractions were markedly lower, with no change in the pD2 values (i.e. the negative logarithm of the concentration for half-maximal response), in cavernosal strips obtained from castrated rabbits than in those from controls. Endothelium-dependent relaxation elicited by carbachol increased in the castrated group but the relaxation induced by sodium nitroprusside did not change and those elicited by adenosine were strongly depressed when compared with controls. There were no significant changes in the pD2 values of agonist-induced relaxation responses in all groups. The relaxation elicited by electrical-field stimulation at lower frequencies increased in strips from castrated rabbits but at higher frequencies were unchanged when compared with controls. Castration induced changes in the relaxation response of cavernosal strips were significantly restored by in vivo testosterone replacement but those induced by phenylephrine were not. CONCLUSION: The lack of testosterone has an effect on the reactivity of the corpus cavernosum, indicating that testosterone has an important role in erectile function by a pre- or post-synaptic action on the corpus cavernosum. PMID- 9202569 TI - Spinal abnormalities in classic bladder exstrophy. AB - OBJECTIVE: To determine the frequency and clinical sequelae of significant spinal malformations in children born with classic bladder exstrophy. PATIENTS AND METHODS: All patients evaluated or treated for classic bladder exstrophy at this institution were reviewed retrospectively. Radiographs or reports pertinent to the spine were retrieved and reviewed with a paediatric radiologist and all vertebral abnormalities categorized. Clinical charts of those with spinal anomalies were reviewed to determine any clinical neurological disorders associated with the radiographic findings. RESULTS: Of 423 patients with classic bladder exstrophy who were identified, 299 had radiographs or reports available for adequate review. Of these, 34 (11%) normal variants, e.g. spina bifida occulta and lumbarization or sacralization of vertebrae, were identified. Abnormalities of spinal curvature were identified in eight patients (2.7%), all with uncomplicated scoliosis. Spinal dysraphism was diagnosed in 12 patients (4%) and included myelomeningocele, lipomeningocele, scimitar sacrum, posterior laminal defects in two or more vertebrae, vertebral fusion and hemivertebrae. The one patient with myelomeningocele had clinical neurological dysfunction, giving an overall incidence of 0.3%. CONCLUSIONS: Spinal anomalies, excluding normal variants, occur in children born with classic bladder exstrophy at a rate of about 6.7%. The incidence of this association is much less than that for cloacal exstrophy. Although rare, neurological dysfunction can occur in the case of spinal dysraphism. Paediatric urologists and neurologists should be aware of this significant difference between patients with classic bladder and cloacal exstrophy to properly diagnose, evaluate and treat the attendant neurological problems. PMID- 9202570 TI - A comparison of methods of repairing the symphysis pubis in bladder exstrophy by tensile testing. AB - OBJECTIVE: To compare the efficacy of several fixation techniques in the reconstruction of diastasis of the symphysis pubis in bladder exstrophy. MATERIALS AND METHODS: The symphysis of 32 pelves removed from piglets about 1 month old were disrupted and repaired using one of eight methods. After repair, each pelvis was tested biomechanically for load-to-failure, stiffness and energy to-failure. The various repair techniques were compared with one another and to a group of six pelves tested intact. RESULTS: Four of the methods tested, including a #2 nylon suture placed through bone in a horizontal mattress arrangement, several loops of #2 nylon suture tied around the pubes, Mersilene tape tied around the pubes, and Mitek G-II suture anchors placed into the superior pubic rami, showed the highest stiffness and load-to-failure. All methods were very weak compared with the intact symphysis; the best load-to-failure (#2 nylon horizontal mattress suture) was less than half of that for intact bone, and the best stiffness (mersilene tape) was less than one-third that for the intact symphysis. CONCLUSION: The repairs varied greatly in the variables tested and those which are most promising merit further investigation to assess methods of improving their performance. PMID- 9202571 TI - The in situ appendix in the Malone antegrade continence enema procedure for faecal incontinence. PMID- 9202572 TI - Reconstruction of the mons venereum using omentum in female patients with epispadias/exstrophy. PMID- 9202573 TI - Markedly delayed linear growth in a child with cloacal exstrophy. PMID- 9202574 TI - Urinoma and urinary ascites secondary to calyceal perforation in neonatal posterior urethral valves. PMID- 9202575 TI - Imperforate hymen as a cause of bladder perforation and intestinal obstruction. PMID- 9202576 TI - Focal orchitis--a diagnostic dilemma. PMID- 9202577 TI - Recurrent epididymo-orchitis in a child secondary to a stone in the seminal vesicle. PMID- 9202578 TI - Varicocele in three siblings: a previously unreported entity. PMID- 9202580 TI - Giant retroperitoneal angiomyolipoma mimicking liposarcoma. PMID- 9202579 TI - Late relapse of metastatic teratoma invading a vertebral body: a combined surgical approach. PMID- 9202581 TI - Retroperitoneal melanocytic schwannoma. PMID- 9202582 TI - Haematuria and vesico-cutaneous fistula after hip surgery. PMID- 9202583 TI - Radiology guidelines. PMID- 9202584 TI - Optimization of sequences for MRI of the abdomen and pelvis. PMID- 9202585 TI - Pictorial review: magnetic resonance angiography of arterial variants at the Circle of Willis. AB - As intracranial MR angiography becomes more widely used and spatial resolution improves, anomalies at the Circle of Willis which have been previously well described on angiographic studies and anatomic dissections will become more frequently appreciated by MR angiography. Recognition of these variants is important to avoid confusion of the anomalies with aneurysms, evaluate collateral pathways in the intracerebral circulation, and enhance pre-operative planning in patients undergoing surgery at the skull base. In this review, we illustrate several of the more common types of anomalies at the Circle of Willis and discuss the possible clinical significance of each. PMID- 9202586 TI - Small pelvic lymph node metastases: evaluation with MR imaging. AB - The purpose of this study was to determine if lymph node asymmetry in small (< 1.0 cm) pelvic lymph nodes was a significant prognostic feature in determining metastatic disease. Two hundred and sixteen patients who presented with pelvic carcinoma underwent magnetic resonance imaging (MRI). They were correlated with pathological findings obtained at surgery. We considered the maximum diameter (MAD) of both round- or oval-shaped suspicious masses seen in the axial plane. Two different cut-off values were determined: lymph node diameter greater than 1.0 cm (criterion 1) and lymph node diameter greater than 0.5 cm with asymmetry relative to the opposite side for lymph nodes ranging from 0.5 cm to 1.0 cm (criterion 2). With criterion 1, MRI had an accuracy of 88%, a sensitivity of 65%, a specificity of 96%, a positive predictive value (PPV) of 88% and a negative predictive value (NPV) of 88% in the detection of pelvic lymph node metastasis. By considering criterion 2, MRI had an accuracy of 85%, a sensitivity of 75%, a specificity of 91%, a PPV of 71% and a NPV of 91%. Normal small asymmetric lymph nodes were present in 5.6% of cases. Normal asymmetry of pelvic lymph nodes is not uncommon. It cannot be relied on to diagnose metastatic involvement in cases of small suspicious lymph nodes. PMID- 9202587 TI - Temporomandibular joint MRI: a 2-D gradient-echo technique. AB - Magnetic resonance imaging (MRI) of the temporomandibular joint (TMJ) has been most commonly performed using spin-echo sequences. Gradient-echo sequences have previously been investigated in the context of 3-D and 'dynamic' (pseudo kinematic) imaging. We have used gradient-echo to improve image quality in static studies with rapid acquisition, high spatial resolution (512 x 512 matrix) and excellent contrast resolution. Using pragmatic methods and a non-randomized study group we have demonstrated a definite advantage to image quality, and thus diagnostic confidence, from the use of a gradient-echo high spatial resolution sequence incorporating split acquisition open mouth views. PMID- 9202588 TI - Perirenal MR high signal--a new and sensitive indicator of acute ureteric obstruction. AB - PURPOSE: This study was carried out to determine the incidence of perirenal magnetic resonance (MR) high signal in acute ureteric obstruction as demonstrated by half-Fourier acquisition single shot turbo spin-echo (HASTE) MR. In addition, we evaluated the sensitivity of this perirenal MR high signal as a predictor of acute ureteric obstruction. MATERIALS AND METHODS: A prospective evaluation of 55 consecutive patients with suspected ureteric obstruction was carried out using the HASTE MR sequence. Images were compared to concurrent IV urography (IVU) or to computed tomography (CT) where these were available. Acute and chronic ureteric obstruction were differentiated by clinical evaluation. RESULTS: Forty one patients had obstructed kidneys. HASTE MR accurately predicted the presence of acute ureteric obstruction in 20/23 (87%) based on presence of perirenal MR high signal. None of these showed evidence of contrast medium extravasation on the concurrent i.v. urogram. 15/18 (83%) chronically obstructed kidneys demonstrated no perirenal high signal on HASTE MR, and the remaining three showed only a trace of perirenal high signal. CT showed perirenal stranding in only 2/8 patients with acutely obstructed kidneys. CONCLUSION: In acute ureteric obstruction, HASTE MR shows perirenal high signal intensity much more commonly than IVU shows extravasation or CT showing perirenal stranding. The origin of this MR signal is uncertain but may represent oedema, lymphatic distension or free fluid from forniceal rupture. HASTE MR can accurately distinguish between acute and chronic ureteric obstruction based on the degree of perirenal high signal. PMID- 9202589 TI - Renal artery stent placement in renal artery stenosis: technical and early clinical results. AB - We report the technical and early clinical results of renal artery stent placement in 29 consecutive patients treated at a single centre over a 30-month period, employing the Palmaz balloon-expandable stent. Of 32 arteries treated, 23 (72%) were atheromatous, ostial stenoses. Immediate technical success was achieved in all 29 patients. Follow-up angiography was performed on 25 patients at 6.7 months (mean) and demonstrated a patient restenosis rate of 16%. All surviving patients were followed up for a minimum of 6 months. Blood pressure control was improved in eight (50%) of hypertensive patients, and renal function improved in seven (33%) and stabilized in six (29%) patients with chronic renal impairment (serum creatinine > 150 mumols/l). Complications occurred in seven (24%) of patients, including one procedure-related death. Our experience indicates that stent placement has an initial high technical success rate in renal artery stenosis and that this patency is maintained at repeat angiography with a low restenosis rate. Renal artery stenting is likely to extend the role of percutaneous renal revascularization especially in atheromatous ostial lesions. A randomized trial will be required to evaluate its role compared with balloon angioplasty. PMID- 9202590 TI - The clinical importance of axillary lymphadenopathy detected on screening mammography. AB - The aim of this study was to determine the incidence and cause of axillary lymphadenopathy detected by screening mammography and to devise a management protocol for this pathology. In a retrospective study of 95,806 consecutive screening mammograms, 37 cases of 'pathological' axillary nodes were identified using two or more of the following criteria: size > 2 cm, replacement of fatty hilum, rounded shape and generalized increased density. In 16 cases with an additional mammographic abnormality, 12 had a mass (10 malignant and two benign) and four had suspicious calcification (all malignant). In 12 of these cases, the lymph nodes showed malignancy (75%). In 21 patients with lymphadenopathy alone on screening, six patients had a known underlying diagnosis and were not recalled from screening. The remaining 15 patients were recalled for further assessment including fine needle aspiration cytology (FNAC). The ultimate diagnosis was benign in 10 cases (48%)--six reactive changes, one healed granulomatous disease, one rheumatoid arthritis, one amyloid and one acute infection--and malignant in 11 cases (52%)--six non-Hodgkin's lymphoma, four metastatic carcinoma and one leukaemia. In conclusion, there is a high incidence of malignant nodal involvement in cases of screen detected lymphadenopathy (62% of cases in our series). We would advise that patients with lymphadenopathy as the sole finding on screening mammography and in whom there is no known underlying cause should undergo FNAC followed by excision biopsy. Fifty per cent of such patients in this study had underlying malignancy. PMID- 9202591 TI - Paravertebral extravasation of contrast medium during lumbar myelography. AB - We report on paravertebral extravasation of water soluble contrast medium seen during lumbar myelography and/or subsequent post myelogram CT in 10 patients. This is an uncommon feature of lumbar myelography occurring in less than 1% of cases in our experience and which has not been previously documented. PMID- 9202592 TI - Just how valuable is double reporting in screening mammography? AB - The double reporting of screening mammograms has become an aim of most United Kingdom screening units although it is not Department of Health policy. It is recognized that mammogram reporting sensitivity improves with experience. This study was designed to assess how valuable double reporting has been in our unit. The data from the first screening round, including the interval cancers which were detected during the subsequent 3 years, were analysed. The reporting sensitivities of the less experienced radiologist improved during the 3 years of the first screening round from 90.6% to 98.9%. For the more experienced radiologist sensitivities ranged from 97.1% to 98.9%. Overall the increased sensitivity from double reporting over single reporting was 1.5% over the best to 4.2% over the worst single reader. With such a relatively small difference between single and double reporting and high individual reporting standards, it is hard to justify the additional resources required for double reporting. PMID- 9202593 TI - Technical report: perirectal abscess drainage--a simple modification in technique using a vascular sheath. AB - Two patients with pelvic abscesses underwent fluoroscopic guided transrectal catheter drainage using a simple modification in standard technique. A 9F vascular sheath allows contrast to be instilled through the side port and the seal avoids any leakage back through the sheath. It also provides a safe, quick and precise needle puncture at the selected site. PMID- 9202595 TI - Case report: thickening of the walls of non-dilated bile ducts. PMID- 9202594 TI - Case report: fulminant Toxoplasma gondii pneumonia in a patient with AIDS. PMID- 9202596 TI - Case report: antenatal sonographic diagnosis of meconium peritonitis and subsequent evolving meconium pseudocyst formation without peritoneal calcification. PMID- 9202597 TI - Colonic mucosal lymphoid hyperplasia and apthoid ulcers in Crohn's disease. PMID- 9202598 TI - Multiple duodenal gastrinomas demonstrated with spiral CT. PMID- 9202599 TI - What are the indications for a carotid duplex scan? PMID- 9202600 TI - Care of the child with a solid organ transplant: what is the role of the generalist? PMID- 9202601 TI - Cardiac transplantation: perspectives on long-term survival. PMID- 9202602 TI - Cardiac troponin I: a new diagnostic gold standard of cardiac injury in children? PMID- 9202603 TI - Autoantibodies in childhood opsoclonus-myoclonus syndrome. PMID- 9202604 TI - Myeloid disorders in infants with Noonan syndrome and a resident's "rule" recalled. PMID- 9202605 TI - Vertical transmission of hepatitis C: to screen or not to screen. PMID- 9202606 TI - Long-term survivors of pediatric heart transplantation: a multicenter report of sixty-eight children who have survived longer than five years. AB - OBJECTIVE: Short-term survival after pediatric heart transplantation is now excellent, but ultimately the efficacy of this procedure will depend on duration and quality of survival. We sought to evaluate the clinical course of long-term survivors of heart transplantation in childhood. METHODS: Patients who had undergone heart transplantation at the university hospitals of Stanford, Columbia, and Pittsburgh between 1975 and 1989 and survived longer than 5 years from transplantation were identified and their clinical courses retrospectively reviewed. RESULTS: Sixty eight children have survived more than 5 years from transplantation, and 60 (88%) are currently alive with a median follow-up of 6.8 years (5 to 17.9 years). Thirteen have survived more than 10 years from transplantation. Renal dysfunction caused by immunosuppressive agents was common, and two patients required late renal transplantation. Lymphoproliferative disease or other neoplasm occurred in 12 patients, but none resulted in death. Coronary artery disease was diagnosed in 13 patients (19%), leading to retransplantation in eight. Death after 5 years was related to acute or chronic rejection in 5 of 8 cases. Two of the deaths were directly related to noncompliance with immunosuppressive medication. All survivors are in New York Heart Association class 1. CONCLUSIONS: Long-term survival with good quality of life can be achieved after heart transplantation in childhood, though complications of immunosuppression remain common. Posttransplantation coronary artery disease is emerging as the main factor limiting long term graft and patient survival. PMID- 9202607 TI - Cardiac troponin I in pediatrics: normal values and potential use in the assessment of cardiac injury. AB - OBJECTIVE: To establish normal values and determine the impact of congenital or acquired heart disease on serum cardiac troponin I (cTnI). METHODS: Concentrations of cTnI were measured in two groups of children. Group A represented ambulatory pediatric patients with no apparent cardiac disease (n = 120) and patients in stable condition with known congenital or acquired cardiac abnormalities (n = 96); group B was composed of patients admitted to intensive care units with normal echocardiograms (n = 16), with abnormal echocardiograms (n = 36), and those with blunt chest trauma who were thought to have cardiac contusions (n = 7). RESULTS: The cTnI concentrations were generally less than 2.0 ng/ml in group A and frequently below the level of detection for the assay (1.5 ng/ml). There was no statistical difference between the two outpatient subgroups (p = 0.66). Nine intensive care patients had cTnI values greater than 2.0 ng/ml. Six of these patients, all with abnormal echocardiograms, had values less than 7.7 ng/ml. All improved and had subsequent normal cTnI concentrations. None of the three remaining patients (two with systemic illness (trauma and sepsis) and one with severe pulmonary hypertension), all with values greater than 8.0 ng/ml, survived. Three of the four patients with high likelihood of cardiac contusion had cTnI concentrations greater than 2.0 ng/ml (including one patient who died). CONCLUSIONS: Cardiac troponin-I values are generally not elevated in children with stable cardiac disease or general pediatric conditions. In the context of severe acute illness, significant elevation of cTnI may be an indicator of poor outcome. Elevation of cTnI may also have diagnostic value in cases when cardiac contusion is suspected. PMID- 9202608 TI - Serum autoantibodies in childhood opsoclonus-myoclonus syndrome: an analysis of antigenic targets in neural tissues. AB - OBJECTIVE: Opsoclonus-myoclonus (OM) is a rare neurologic syndrome affecting children and adults. In children it occurs as a parainfectious process or a paraneoplastic syndrome in association with neuroblastoma. Evidence for an immune mechanism includes the presence of serum autoantibodies to several neural antigens and improvement of symptoms with immunosuppressive therapy. We studied the neural antigenic targets of serum IgM and IgG autoantibodies from nine children with OM. DESIGN: We studied sera from nine children with OM, three with associated neuroblastoma and six with a prodromal viral illness. Control subjects (n = 77) included four children with neuroblastoma but not OM, 32 children with other neurologic disorders, and 41 with nonneurologic illnesses. We studied the neural antigenic targets of serum IgM and IgG autoantibodies by the following methods: (1) immunostaining of human cerebellar sections and peripheral nerve, and (2) Western blot analysis with human brain fractions including white matter, gray matter, and cerebellar Purkinje cells and nuclei. RESULTS: Sera from all nine children with OM had IgM and IgG binding to the cytoplasm of cerebellar Purkinje cells and to some axons in white matter. In peripheral nerve, IgM and IgG from all nine OM sera bound to large and small axons. Western blot analysis showed a distinctive pattern of binding to several neural proteins, including a 210 kd antigen identified as the high molecular weight subunit of neurofilament. No control serum showed a similar pattern of reactivity. CONCLUSION: Opsoclonus myoclonus syndrome in childhood is associated with a distinctive pattern of serum IgM and IgG binding to neural tissues and antigens. PMID- 9202609 TI - Occurrence of myeloproliferative disorder in patients with Noonan syndrome. AB - We report four cases of Noonan syndrome associated with chronic myelomonocytic leukemia in childhood. These children shared some hematologic features: thrombocytopenia, splenomegaly in the first months of life, occurrence of chronic myelomonocytic leukemia without abnormalities of the initial bone marrow karyotype, and, in three cases, improvement of the hematologic disease. A common pathophysiologic process in such patients is suggested. PMID- 9202610 TI - Effect of maternal CD4+ cell count, acquired immunodeficiency syndrome, and viral load on disease progression in infants with perinatally acquired human immunodeficiency virus type 1 infection. New York City Perinatal HIV Transmission Collaborative Study Group. AB - Among a cohort of 152 infants perinatally infected with human immunodeficiency virus type 1, and their mothers, we correlated infant outcome with material CD4+ lymphocyte count and the presence of maternal acquired immunodeficiency syndrome near delivery. In a subset of 50 mother-infant pairs, we also correlated infant outcome with maternal quantitative viral burden as measured by the nucleic acid sequence based amplification system. We found that low maternal CD4+ cell count and high viral burden were associated with decreased time to category C disease or death in infants infected with human immunodeficiency virus type 1. In a multivariate analysis, high maternal viral load and maternal acquired immunodeficiency syndrome were independently associated with shorter time to category C disease or death in infants with human immunodeficiency virus type 1 infection. High viral load in pregnant women, independent of the presence of advanced maternal disease, appears to increase the risk of rapidly progressive disease in their infected offspring. PMID- 9202611 TI - Correlation of ribonucleic acid polymerase chain reaction, acid dissociated p24 antigen, and neopterin with progression of disease. A retrospective, longitudinal study of vertically acquired human immunodeficiency virus type 1 infection in children. AB - OBJECTIVE: We investigated the relationship between cell-free viral load, neopterin, age-adjusted CD4+ cell concentration, and clinical events in 49 children with vertically acquired human immunodeficiency virus type 1 infection. STUDY DESIGN: Viral load was measured by quantitating viral ribonucleic acid in serum by polymerase chain reaction and measurement of immune complex dissociated p24 antigen in serum and plasma. Children were followed for an average of 2 1/2 years, with an average of 6 samples per child. Medical records were reviewed for weight, CD4+ cell count and clinical events. RESULTS: High virus copy number in serum was predictive of a decrease in weight-for-age zscore during the subsequent 6 months. High viral load, low CD4+ cell count, and high neopterin level were correlated with encephalopathy. High viral load correlated with opportunistic infections. All of these relationships held regardless of treatment status, although viral load decreased significantly after treatment was begun. CONCLUSIONS: Measurements of viral load were useful prognostic indicators for poor weight gain. Elevated serum virus levels and neopterin values and low CD4+ cell counts were all associated with encephalopathy. PMID- 9202612 TI - Zidovudine administered to women infected with human immunodeficiency virus type 1 and to their neonates reduces pediatric infection independent of an effect on levels of maternal virus. AB - OBJECTIVE: To determine whether zidovudine, administered to reduce vertical transmission of human immunodeficiency virus type 1 (HIV-1), impacts the level of maternal viral DNA within the lymphocytes of infected pregnant women. STUDY DESIGN: A prospective, nonrandomized study of 42 HIV-1 infected pregnant women. Nineteen women received zidovudine therapy to reduce HIV-1 perinatal transmission, and 23 were untreated. HIV-1 DNA was determined by polymerase chain reaction amplification of lymphocyte DNA from maternal blood samples obtained at the time of delivery. Treated and untreated, transmitting and nontransmitting groups were compared for clinical, virologic, and immunologic parameters with at test or a Fisher Exact Test, and for copies of HIV-1 DNA per 10(6) CD4+ T cells with a Mann-Whitney rank sum test. RESULTS: Untreated pregnant women who transmitted HIV-1 to their infants had tower CD4+ T-cell counts and a greater degree of immune complex dissociated p24 antigenemia than did the untreated nontransmitting group (p < 0.01) but did not differ significantly with respect to age, race, or mode of delivery. The level of HIV-1 proviral DNA within lymphocytes was significantly greater in the untreated transmitting group than in the nontransmitting mothers (p = 0.003). Zidovudine treatment resulted in a 78% decrease in maternal transmission (p = 0.017). However, there was not a significant difference in DNA copy numbers in CD4+ T cells in the treated compared with the untreated groups. CONCLUSION: Zidovudine reduces HIV-1 maternal transmission independent of its effect on the level of the maternal peripheral blood proviral load. PMID- 9202613 TI - Impaired early growth of infants perinatally infected with human immunodeficiency virus: correlation with viral load. AB - OBJECTIVE: To evaluate the effect of viral load on the early growth of infants infected with human immunodeficiency virus (HIV). METHODS: Plasma concentrations of p24-antigen and HIV ribonucleic acid were measured retrospectively and correlated with growth parameters for the first 18 months of life in a cohort of 47 term infants born to HIV-infected mothers prospectively enrolled in a study of perinatal HIV transmission. Comparisons of the mean weight and length of the 18 HIV-infected and 29 uninfected infants for each interval and across intervals were made. Viral load was correlated with standard deviation scores. Infants were stratified by high and low viral load during the first 6 months of life. RESULTS: At birth, no difference in weight and length was observed between HIV-infected and uninfected infants. Between birth and 6 months of age, the infected infants grew less rapidly than the uninfected infants, a finding temporally associated with an exponential increase in HIV viremia. The linear growth of infected infants remained consistently less than that of the uninfected infants after 6 months of life, although the differences were no longer statistically significant and tended to decrease with age in parallel with declines in viral load. The median plasma concentration of HIV ribonucleic acid was significantly higher at 3, 6, 12, and 18 months in infected infants in whom growth failure developed. Infants who had a high viral load in the first 6 months of life were significantly more likely to have severe growth failure. Though the mean SD for weight of the infected infants was always less than that of the uninfected infants, the differences were small and not significant. CONCLUSIONS: Our results confirm the observation that stunting is an early frequent finding in perinatal HIV infection. The deleterious effect of HIV on linear growth appears to be correlated with the level of postnatal HIV viremia, although the exact mechanism of this association remains to be elucidated. PMID- 9202614 TI - Decline of pediatric admissions with Haemophilus influenzae type b in New York State, 1982 through 1993: relation to immunizations. AB - OBJECTIVES: To evaluate the impact of vaccination for Haemophilus influenzae type b (Hib) on pediatric hospital admissions in New York State, and to identify risk factors in children who continue to be admitted for Hib invasive disease. METHODS: Retrospective review of hospitalizations in New York state from 1982 through 1993 and a survey of immunization records from physician offices in Monroe Country, New York. RESULTS: In 1982, 769 children were admitted to New York state hospitals for Hib-related conditions; by 1993, this had decreased to 133. Significant declines during the study period occurred in the age-adjusted admission rates for Hib meningitis, septicemia, pneumonia, and epiglottitis, but not for arthritis and osteomyelitis. In 1993 alone, 712 admissions, 18 deaths, and 135 episodes of morbidity were avoided. Since 1991, the rates of admissions for Hib-related conditions have remained fairly constant. Minority subjects continue to be twice as likely as white subjects to be admitted for invasive Hib disease (0.44 vs 0.17/100,000). Children living in urban Rochester also are more likely to be admitted and less likely to be completely immunized against Hib (61%) than those living in suburban areas (82%). CONCLUSIONS: Although Hib vaccine has had a major impact on hospital admissions for Hib-related conditions, the goal of completely eliminating Hib disease will require programs targeted at groups at high risk, such as minorities and those living in cities. PMID- 9202615 TI - Changes in gastric emptying in early postnatal life. AB - OBJECTIVE: To study the pattern of gastric emptying in very premature infants and to determine whether there are changes with postnatal age and the ability to tolerate feedings. METHODS: Sequential ultrasound measurements of the gastric antral cross-sectional area were obtained in 32 infants (mean gestational age, 26 +/- 1 weeks) before and after feeding for 2 hours. Studies were carried out after initiation of feedings, when full feedings were received, and at 32 weeks. Infants classified as feeding intolerant (n = 9) were also studied when feedings were restarted. Gastric emptying was assessed by the time taken for antral cross sectional area to reach maximal value and to decrease to half the maximal increment (half-antral clearance). RESULTS: Delayed antral distention was observed at the time of the initial study in both feeding-tolerant (8 of 23) and feeding-intolerant (8 of 9) infants; however, there were significant differences in times for maximal antral distention (p < 0.002) and half-antral clearance (p < 0.006) between the feeding-tolerant and feeding-intolerant infants. By the time of full feedings, the feeding-intolerant infants showed immediate gastric emptying but still had a longer half-antral clearance time (p < 0.01). By 32 weeks, all infants had immediate antral distention and a more mature curvilinear pattern of gastric emptying. CONCLUSIONS: Knowledge of these different patterns of gastric emptying in very premature infants may lead to the development of more rational feeding strategies. PMID- 9202617 TI - Eosinophil cationic protein in tracheal aspirates of preterm infants with bronchopulmonary dysplasia. AB - Eosinophil cationic protein was elevated during the first week of life in tracheal aspirates from 11 preterm infants in whom bronchopulmonary dysplasia subsequently developed compared with 8 preterm and 8 term infants without bronchopulmonary dysplasia. Eosinophil cationic protein levels increased progressively with continued intubation in the infants with bronchopulmonary dysplasia but remained low in a comparison group of term infants. We suggest that eosinophils participate in the inflammatory process in bronchopulmonary dysplasia and may contribute to lung injury. PMID- 9202616 TI - Effect of physical training on heart-period variability in obese children. AB - OBJECTIVE: The beat-to-beat variability in electrocardiogram intervals (RR, i.e., heart-period variability) provides information on cardiac autonomic activity that predicts arrhythmias and mortality rate in animals and adults. We determined the effect of physical training on heart-period variability in obese children. METHODS: Thirty-five subjects were randomly assigned to physical training and control groups. The training involved 4 months of exercise, 5 days per week, 40 minutes per day. Cardiovascular fitness was measured with submaximal heart rate during supine cycling; percentage of body fat was measured with dual-energy absorptiometry; and resting heart-period variability parameters were measured in a supine position. A pretraining to posttraining change score was computed for each variable. The effect of the training was determined by comparing the changes of the training and control groups. RESULTS: Compared with the control group, the trained group (1) reduced submaximal heart rate and percentage of body fat (p < 0.01); (2) increased in the root mean square of successive differences, a time domain parameter reflective of vagal tone (p < 0.05); (3) decreased in low frequency power expressed as a percentage of total power, a frequency-domain index of combined sympathetic and vagal activity (p < 0.03); and (4) decreased in the ratio of low- to high-frequency power, an index of sympathetic parasympathetic balance (p < 0.01). CONCLUSIONS: In obese children, physical training alters cardiac autonomic function favorably by reducing the ratio of sympathetic to parasympathetic activity. PMID- 9202618 TI - Insulin resistance is associated with decreased clinical status in cystic fibrosis. AB - Patients with cystic fibrosis (CF) frequently have impaired glucose tolerance and progression to diabetes (DM) with clinical features of both insulin-dependent and non-insulin-dependent diabetes. One feature of non-insulin-dependent DM is decreased insulin sensitivity, also known as insulin resistance. The goal of this study was to determine whether patients with CF exhibit insulin resistance and to determine the potential effect of insulin resistance on clinical status. We also sought to determine whether insulin resistance is associated with a specific CF genotype. We studied 18 patients with CF (8 with normal glucose tolerance, 5 with impaired glucose tolerance, 5 with DM), and 20 lean control subjects matched for age, weight, and sex. All control subjects had normal glucose tolerance. The clinical status for each CF patients was determined according to a modified National Institutes of Health scoring system. Each subject underwent a three-step hyperinsulinemic euglycemic clamp (insulin doses of 10, 40, 120 mU/m2 per minute). Results from the 120 mU/m2 per minute infusion defined maximal glucose disposal rate (defined in milligrams per kilogram body weight per minute) at steady state with peripheral insulin levels 195 +/- 20 mU/ml. Subjects with CF demonstrated insulin resistance (control subjects = 13.6 +/- 1.1, patients with CF = 10.2 +/- 1.6 mg/kg per minute; p = 0.003). When each subgroup was compared separately with control subjects, all subgroups were statistically insulin resistant (glucose disposal rate, patients with CF and normal glucose tolerance = 10.8; those with impaired glucose tolerance = 8.4; those with DM = 10.1 mg/kg per minute), and the patients with CF with impaired glucose tolerance were the most insulin resistant. When plotted versus glucose disposal rate, a striking positive correlation between worsened clinical status and insulin resistance (r = 0.85) is demonstrated. Furthermore, there is no correlation between insulin resistance and fasting blood glucose, subject age, or percent ideal body weight (all r values not significant). In conclusion, patients with CF exhibit insulin resistance that is associated with worsened clinical status. We believe it is the combination of insulin resistance and decreased insulin secretion that is responsible for the high incidence of CF-related diabetes. PMID- 9202619 TI - Screening for hereditary spherocytosis by use of automated erythrocyte indexes. AB - OBJECTIVE: To determine whether the mean corpuscular hemoglobin concentration (MCHC) or other erythrocyte indexes, as determined by automated cell counters, remains a useful screening test for identifying patients with hereditary spherocytosis (HS). METHODS: Erythrocyte indexes from 112 children with HS who had not undergone splenectomy were compared with those measured in an equal number of healthy, age-matched children. All indexes were derived from measurements obtained by aperture impedance. RESULTS: Mean corpuscular hemoglobin concentration in the HS group was 35.9 gm/dl, significantly higher than in normal control subjects (34.3 gm/dl; p < 0.001). Mean erythrocyte distribution width also was significantly higher in patients with HS (19.3 vs 12.6; p < 0.001). The MCHC distinguishes individuals with HS, with an area under the receiver operating characteristic curve of 0.86. Although not disease specific, an erythrocyte distribution width > 14 has 85% sensitivity and 97% specificity and an area under the receiver operating characteristic curve of 0.92. An MCHC > 35 gm/dl has a sensitivity of 70% and a specificity of 86%. Combining the MCHC and erythrocyte distribution width increases the area under the receiver operating characteristic curve to 0.97. Specificity is 100% and likelihood ratio is infinite when both the MCHC and erythrocyte distribution width are elevated. CONCLUSIONS: The automated MCHC is an effective screening test to identify children with HS. An elevated erythrocyte distribution width adds additional specificity and is itself a powerful screening tool. The combination of the two tests is an excellent predictor for the diagnosis of HS. PMID- 9202620 TI - Acute splenic complications in children with sickle cell-hemoglobin C disease. AB - OBJECTIVE: To determine the frequency and severity of acute splenic complications in children and adolescents with sickle cell (SC) hemoglobin C disease. METHODS: The medical records of 271 patients with SC disease seen at our center were reviewed to evaluate the incidence and severity of acute complications involving the spleen. RESULTS: Sixteen (6%) children had acute splenic complications. Thirteen (5%) had 16 episodes of acute splenic sequestration (ASSC), with the initial event occurring at a mean age of 8.9 years (range, 2 to 17 years). Splenomegaly had been noted before the initial event in 6 (46%) of the 13 cases, and 3 (23%) had a history of painful splenic infarction. Two young children (aged 4 and 6 years) had a hemoglobin value less than 2 gm/dl, one without history of splenic enlargement. Three (23%) children had a second episode of ASSC. Three additional patients had a history of acute painful splenic infarction, two of whom also had splenic hemorrhage. Eight (3%) of the 271 children required splenectomy (1 after the initial episode of ASSC, 3 after a second episode of ASSC, 2 as a result of pain accompanying chronic infarction and ASSC, and 2 because of splenic hemorrhage). No deaths resulted from ASSC. CONCLUSIONS: We conclude that (1) acute splenic complications in children and adolescents with SC disease are relatively uncommon, (2) most episodes of ASSC occur in preadolescents, (3) ASSC can be life threatening, even in younger children, and (4) prior splenomegaly is not a good predictor of ASSC. Thus it is vital that the parents of all children with SC disease be instructed to palpate their child's spleen regularly. PMID- 9202621 TI - Comparison of dimercaptosuccinic acid and calcium disodium ethylenediaminetetraacetic acid versus dimercaptopropanol and ethylenediaminetetraacetic acid in children with lead poisoning. AB - OBJECTIVES: To compare the response to dimercaptopropanol (BAL) and calcium disodium ethylenediaminetetraacetic acid (EDTA) versus orally administered meso 2,3-dimercaptosuccinic acid (DMSA) and EDTA in children with lead poisoning. METHODS: Retrospective review of medical records of children admitted to MetroHealth Medical Center with a whole blood lead (BPb) concentration of 2.17 mumol/L (45 micrograms/dl) or more (or less than 2.17 mumol/L and not a candidate for outpatient oral chelation) and treated with BAL + EDTA or DMSA + EDTA. In each group, the mean BPb values at the end of therapy and at 14 and 33 days after chelation were compared with pretreatment BPb by the Wilcoxon signed-rank test, whereas the Mann-Whitney U test was used to compare percentage change from pretreatment at each follow-up day between the two groups. RESULTS: Twenty-three children received BAL + EDTA and 22 received DMSA + EDTA. The BPb values (mean +/ SD) at the end of therapy and at 14 and 33 days after chelation were significantly lower than pretreatment in both groups (BAL + EDTA: 17 +/- 10, 34 +/- 7, 36 +/- 11 vs 58 +/- 14 micrograms/dl, p < 0.02, 0.01, 0.001, respectively; DMSA + EDTA: 10 +/- 4, 30 +/- 10, 30 +/- 14 vs 50 +/- 10 micrograms/dl, p < 0.01, 0.001, 0.01, respectively). The percentage reduction (mean +/- SD) in BPb from pretreatment at the end of therapy and on days 14 and 33 after chelation did not differ between the groups (BAL + EDTA: -71.2% +/- 19.8%, -40.2% +/- 13.8%, -37.1% +/- 17%; DMSA + EDTA: -79.9% +/- 8.7%, -38.3% +/- 21.6%, -37% +/- 32%; p > 0.20). Elevation of alanine aminotransferase and vomiting during therapy were observed more frequently in the BAL + EDTA group compared with the DMSA + EDTA group. CONCLUSIONS: Treatment with DMSA or BAL combined with EDTA results in a comparable reduction in BPb. PMID- 9202622 TI - Black children deficient in galactose 1-phosphate uridyltransferase: correlation of activity and immunoreactive protein in erythrocytes and leukocytes. AB - A recent study found a high prevalence of a missense mutation (S135L) in the gene for galactose 1-phosphate uridyltransferase (GALT) in black children with galactosemia (J Pediatr 1996; 128:89-95). In the present study, GALT activity and GALT protein content were measured in erythrocytes and leukocytes of eight black and seven white galactosemic (GALT-deficient) children, for correlation with the presence of the S135L and Q188R (highly prevalent in white galactosemic children) missense mutations. The S135L mutation was found in 9 of 16 alleles of black children but not in white children; the Q188R mutation was found in 10 of 14 alleles examined in white galactosemic children and in 4 of 16 alleles in black galactosemic children. The GALT activity was near zero in the erythrocytes of white and black galactosemic children (0.26 +/- 0.28 vs 0.33 +/- 0.25 mumol/hr per gram of hemoglobin, respectively; p = 0.61) (normal 17 to 26 mumol/hr per gram), and no correlation of erythrocyte activity with genotype was observed. The GALT activity was higher in the leukocytes of black galactosemic children compared with white children (5 +/- 6 vs 1 +/- 2 mumol/hr per gram, respectively) (normal 172 to 374 mumol/hr per gram), but the difference was not statistically significant (p = 0.11). Analysis by genotype revealed that the two S135L homozygotes had much more leukocyte activity (9 and 17 mumol/hr per gram) than Q188R homozygotes or than all non-S135L allelic genotypes. Compound heterozygotes (S135L/G) had intermediate activity. The GALT protein was not detectable by Western blot in the erythrocytes of either white or black galactosemic children, as determined by antibodies specific for both C- and N-terminal sequences. The GALT protein was undetectable in the leukocytes of white galactosemic children, but leukocytes from black galactosemic children with the S135L mutation contained reduced but readily detectable GALT protein. Erythrocyte galactose 1-phosphate levels were significantly lower in galactosemic children with an S135L mutant allele (1.1 +/- 0.2 gm/dl) compared with children who had other mutations (3.1 +/ 0.9 mg/dl; p = 0.0001). The correlation of protein content data with activity levels in the blood cells suggests that the S135L missense mutation affects the stability of GALT protein to produce a deficiency state. PMID- 9202623 TI - A simplified schedule to integrate the hepatitis B vaccine into an expanded program of immunization in endemic countries. AB - OBJECTIVE: To investigate the safety, immunogenicity, and efficacy of a simplified hepatitis B vaccination schedule. METHODS: The second dose of hepatitis B vaccine and the first dose of diphtheria-tetanus-pertussis (DTP) vaccine were given simultaneously at age 6 weeks. The second dose of DTP vaccine was given at age 3.5 months. The third dose of DTP vaccine and the third dose of hepatitis B vaccine were given at age 5.5 months. One hundred three infants (group A) born to mothers without hepatitis B surface antigen (HBsAg) received DTP with whole-cell pertussis vaccine. Fifty-five infants (group B) born to mothers with HBsAg and hepatitis B e antigen received DTP with acellular pertussis vaccine. RESULTS: By age 9 months, none of group A and 4 (7%) group B infants were sero-positive for HBsAg. The protective efficacy against the hepatitis B carrier state in these infants at high risk was 92%. Antibody to hepatitis B surface antigen was 10 mlU/ml or greater in 99 (96%) of group A infants and in 50 (91%) of group B infants. Both the acellular and whole-cell DTP vaccines were immunogenic, and the incidences of adverse reactions were within an expected and acceptable range. CONCLUSIONS: The simplified vaccination schedule to integrate the hepatitis B vaccine into the Expanded Programme of Immunization was safe, Immunogenic, and effective. This schedule may improve vaccine compliance and be applied to DTP and hepatitis B combination vaccines now under investigation. PMID- 9202624 TI - Immunity to diphtheria and tetanus in a young population on a dialysis regimen or with a renal transplant. AB - In 54 transplant recipients diphtheria and tetanus immunity after primary vaccination was significantly lower than in 57 control subject and 35 patients on a dialysis regimen. After a booster, tetanus antibodies developed in the transplant recipients and dialysis patients but no diphtheria antibodies developed in two transplant recipients. No adverse reactions, including acute graft rejection episodes, occurred. PMID- 9202625 TI - Hepatitis C virus infection and related liver disease in children of mothers with antibodies to the virus. AB - OBJECTIVE: To evaluate the clinical, biochemical, and virologic features associated with hepatitis C virus (HCV) infection acquired early in life from mothers with antibodies to HCV (anti-HCV). STUDY DESIGN: Multicenter prospective retrospective study in Italian children. PATIENTS: Two groups of children were investigated. Group 1 included 14 infants, born to mothers with anti-HCV but without human immunodeficiency virus infection, who became seropositive for HCV RNA during the first year of life and were thus considered infected. Group 2 included 16 children with chronic hepatitis C, aged 1 1/2 to 14 years, whose mothers were the unique potential source of infection. Both groups were followed for 12 to 48 months. METHODS: Alanine transaminase (ALT), anti-HCV, and HCV RNA were investigated by the polymerase chain reaction on entry to the study and during follow-up. RESULTS: All children in group 1 had anti-HCV throughout follow up, and all had ALT abnormalities, ranging from 1.5 to 10.5 times the normal value during the first 12 months. During further follow-up, 5 of 10 children had HCV RNA with abnormal ALT values, 3 had a return to normal of the ALT values but continued to have viremia, and 2 eventually had normal ALT values and clearance of HCV RNA. Of the 16 children in group 2, all were free of symptoms and 62% had only slight ALT elevations; 7 who underwent liver biopsy had histologic features of minimal or moderate hepatitis. CONCLUSIONS: HCV infection acquired early in life from mothers with anti-HCV is usually associated with biochemical features of liver damage during the first 12 months of life. Progression to chronicity seems to occur in the majority of cases, although HCV-associated liver disease is likely to be mild throughout infancy and childhood. PMID- 9202626 TI - Human immunodeficiency virus type 2 infection in children. AB - Human immunodeficiency virus type 2 infection is rare in children. This virus can be acquired through transfusion and also by the maternofetal route, especially when the mother becomes infected during pregnancy. Diagnosis based on specific serologic tests is simple after the age of 18 months. In the perinatal period, however, viral isolation by culture or polymerase chain reaction DNA amplification or both appears to be less sensitive than in the case of human immunodeficiency virus type 1. Disease progression is far slower than with human immunodeficiency virus type 1, but severe immunodeficiency can occur. PMID- 9202627 TI - Transient lupus anticoagulants associated with hemorrhage rather than thrombosis: the hemorrhagic lupus anticoagulant syndrome. AB - Lupus anticoagulants (LAs) represent a diverse group of antibodies directed against phospholipids. Patients with LAs may be free of symptoms but can have thrombotic complications including stroke, placental infarction, and fetal loss. Rarely hemorrhagic symptoms have been reported. We describe six previously healthy children who were first seen with clinical bleeding and prolonged activated partial thromboplastin time. Laboratory evaluation revealed positive results on mixing studies and evidence of phospholipid dependence of the anticoagulant, suggesting LAs. Four of six patients had anticardiolipin antibodies, and all four who were tested had reduced factor II activity levels. In all patients, bleeding symptoms resolved spontaneously within 3 months, and laboratory findings returned to normal within 6 months. The hemorrhagic LA syndrome should be considered in previously healthy children with new-onset bleeding and prolonged activated partial thromboplastin time. This clinical entity probably represents pathogenic mechanism distinct from thrombotic LA syndromes. PMID- 9202628 TI - Thrombocytopenic purpura and anemia in a breast-fed infant whose mother was treated with valproic acid. AB - Valproic acid is an antiepileptic drug in widespread use. The possibility of various hematologic side effects with this drug is well recognized. We describe a breast-fed infant with thrombocytopenic purpura, anemia, and reticulocytosis, whose mother was treated with valproic acid. As the mother stopped breast feeding, the infant recovered. PMID- 9202629 TI - Developmental profile of mitochondrial glycine N-acyltransferase in human liver. AB - OBJECTIVE: To study the developmental profile of glycine N-acyltransferase (GAT) in the livers of children of various ages and to compare the total and specific GAT activity with that of the adult control subjects. METHODS: We measured the specific and the total mitochondrial activity of GAT in liver samples taken from 13 children 4 hours to 11 years of age. The samples were compared with those of control adults aged 24 to 40 years. Samples, either from liver-transplant donors or from autopsy, from those who died of a disorder not related to the liver, were obtained between 6 and 36 hours after death. RESULTS: At 4 hours after birth, very low specific activity and the total liver mitochondrial activity were observed (0.19 mumol/min per milligram protein and 210 mumol/min), with a steady increase up to age 7 months (2.51 mumol/min per milligram protein and 812 mumol/min). The mean specific and total GAT activity in children (n = 5) aged 18 months to 11 years was 6.38 +/- 0.13 and 1389 +/- 43 and in control adults aged 24 to 40 years (n = 3) was 6.5 +/- 0.3 and 1461 +/- 71 mumol/min per milligram protein and mumol/min, respectively. These specific and total GAT activity values from children aged 18 months to 11 years were not statistically significant (by analysis of variance and Mann-Whitney test) in comparison with the corresponding activity values from the adult control subjects. CONCLUSIONS: Our results indicate that up to age 7 months, children have only 5% to 40% of liver GAT specific activity, whereas the peak activity is achieved at 18 months and remains constant until age 40 years. The delayed development of GAT in children may thus compromise the detoxification of various drugs and xenobiotics. PMID- 9202630 TI - Is it reactivation of fetal hemoglobin synthesis after transplantation of cord blood stem cells from a donor with heterozygous sickle cell anemia or beta thalassemia? PMID- 9202631 TI - Cryptosporidium in patients infected with human immunodeficiency virus: azithromycin revisited. PMID- 9202632 TI - Excessive playing of home computer games by children presenting unexplained symptoms. PMID- 9202634 TI - Confusion re: nipple confusion. PMID- 9202633 TI - Differences in the in vivo insulin secretion and sensitivity of healthy black versus white adolescents. PMID- 9202635 TI - Cyclosporine in activated macrophage and histiocytic syndromes. PMID- 9202636 TI - "Automatic" diagnosis of viral enteritis. PMID- 9202637 TI - Low birth weight, preterm delivery, and periconceptional vitamin use. PMID- 9202638 TI - Ward evaluations: should they be abandoned? AB - Even in the era of the objective structured clinical examination (OSCE), the predominant method of resident evaluation is the faculty ward evaluation (WE), despite many concerns about its reliability. The aim of this study was to determine the value of the WE as a measurement of clinical competence in terms of both reliability and validity. In a one-year period, surgery faculty members evaluated 72 residents. An average of 7 faculty members evaluated each resident. The evaluation form contained 10 specific performance ratings and an overall evaluation. Inter-rater reliability of the overall performance ratings was calculated by using the intraclass correlation. Validity of the WE was evaluated in four ways. Inter-rater reliability of the overall performance rating was 0.82; the reliability of a single overall rating was 0.39. (1) A discriminant function analysis indicated that residents at advanced levels of training received more positive evaluations than residents at less advanced levels (P < 0.0001). (2) The overall rating was significantly correlated (r = 0.55, P < 0.0001) with the overall score of a concurrent OSCE. (3) A factor analysis showed high correlations among the items, indicating a lack of discrimination between the skills. (4) Overall ratings were insensitive to performance deficiencies. Only 1.3% of the ratings were unsatisfactory or marginal. The WE was sufficiently reliable to estimate the faculty's view of each resident. The fact that the ratings tended to differentiate residents by level of training and that ratings significantly correlated with the OSCE provides strong evidence of their validity. However, factor analysis indicated that the faculty members were making one global, undifferentiated judgment and that these ratings did not identify deficient performance skills. We conclude that ward evaluations have a place in the assessment of residents. PMID- 9202639 TI - Immunodepressive effects of LPS on monocyte CD14 in vivo. AB - Having previously reported that septic patients displayed lower levels of monocyte CD14 (endotoxin receptor) as compared to normal individuals, we were interested in the hypothesis that lipopolysaccharide (LPS) modulates levels of monocyte CD14 in vivo. We examined CD14 expression in 13 human volunteers who were given a non-lethal injection of Escherichia coli LPS (4.0 ng/kg). Monocyte CD14 was assayed by direct immunofluorescent determination with appropriate anti CD14 monoclonal antibodies using flow cytometry. To test for cell responsiveness, monocytes were additionally examined following in vitro stimulation by phorbol myristate acetate (PMA) and N-formyl-methionyl-leucyl-phenylalanine (FMLP). Following LPS infusion, all patients displayed significant monocytopenia and responded with fever and tachycardia. Plasma samples demonstrated elevated levels of TNF alpha. CD14 expression was down-regulated by 52% on monocytes obtained 3 hr following LPS infusion (P < 0.05, vs. pre-LPS levels). Monocytes obtained pre LPS infusion were down-regulated following in vitro stimulation by PMA to levels 72 +/- 8% and by FMLP to levels 75 +/- 5% of unstimulated control cells. In contrast, monocytes obtained 3 hr post-LPS infusion failed to respond to PMA or FMLP with significant down-regulation. LPS down-regulated CD14 expression on monocytes in vivo and LPS also blunted the ability of monocytes to respond to other stimuli. We conclude that LPS desensitizes monocytes to itself and thereby renders an immunodepressive effect on these cells. PMID- 9202640 TI - Endotoxin-induced, neutrophil-mediated endothelial cytotoxicity is enhanced by T lymphocytes. AB - The role of T-lymphocytes (T cells) and interferon-gamma (IFN-gamma) in the pathogenesis of sepsis-induced microvascular endothelial injury remains unclear. We sought to determine whether the syngeneic coculture of human T cells in the presence of LPS promoted subsequent neutrophil (PMN)-mediated endothelial cytotoxicity. Syngeneic T cells were cocultured with 51Cr-loaded human adipose microvascular endothelial cell (HAMVEC) monolayers in the absence and presence of LPS. Subsequent PMN-mediated HAMVEC cytotoxicity (measured as percent specific 51Cr release) was absent in cultures that contained T cells but no LPS and was significantly increased when T cells were cocultured in the presence of LPS. This was true both following addition of unstimulated PMNs (-0.8 +/- 3.0% vs 4.9 +/- 4.7% for T cells alone vs T cells plus LPS, respectively) and PMNs stimulated with f-Met-Leu-Phe (-0.4 +/- 3.1% vs 10.7 +/- 3.0% for T cells alone vs T cells plus LPS, respectively). Increased cytotoxicity was associated with increased expression of the endothelial adhesion molecules ICAM-1 and VCAM-1. Control experiments failed to demonstrate cytotoxicity when HAMVEC were cultured in the presence of IFN-gamma alone, LPS alone, or T cells without LPS. It appears that there is a necessary requirement of both LPS and (presumably activated) T cells or their products (other than IFN-gamma) for enhanced PMN-mediated endothelial cytotoxicity. This phenomenon may also be mediated by increased expression of endothelial adhesion molecules that promote subsequent PMN adhesion. PMID- 9202642 TI - Adenoviral transfection of isolated pancreatic islets: a study of programmed cell death (apoptosis) and islet function. AB - Gene therapy provides a potential technique to modify immunity in vitro and therefore may prolong graft survival in vivo. However, viral infection and gene transfer may damage target cells and interfere with biologic function. Viruses, including adenovirus, are known to be capable of modulating apoptosis and initiating cell death by either inducing or suppressing specific processes, depending on the virus and cell system studied. The effect of adenovirus on islet cell viability and function has not been examined in detail. In this study, the dose-dependent effect of an adenoviral vector on islet cell death and glucose stimulated insulin secretion (GSIS) was investigated to establish a therapeutic window for the dose of viral vector administered. Isolated pancreatic rat islets were incubated with an adenovirus expressing a beta-galactosidase gene (AdHCMVsp1LacZ) at different viral concentrations [multiplicity of infection (MOI) 1:10, 1:100, and 1:1000]. Transfection rate, in vitro and in vivo islet viability, and occurrence of programmed cell death were determined 1, 3, and 7 days after transfection. Islets, transfected at MOI 1:10 and 1:100, demonstrated apoptosis not significantly different from nontransfected controls. Islets, transfected at MOI 1:1000, demonstrated a significant increase in apoptosis at 24 hr, which decreased over 7 days of culture. The increase in apoptosis was not reflected by a significant decrease in in vitro GSIS of surviving islet cells, as assessed by stimulation index following in vitro perifusion. SCID or nude mice transplanted with AdlacZ-transfected islets (MOI 1:100 and 1:1000) remained normoglycemic for > or = 30 days. These results demonstrate that transfection of islets using adenoviral vectors can be manipulated such that efficient expression of the gene product encoded by the transfected gene (beta-galactosidase) can be achieved at lower transfecting concentrations of the adenoviral vector (MOI 1:10, 20.2%; MOI 1:100, 30.7%) while preserving islet function. This efficiency of transfection may allow pretransplant manipulation of isolated islet cells without vector-specific alteration of islet function. In cases where high virus concentrations are required for efficient gene transfer (adequate expression of the transgene product), a deleterious effect of the vector on islet cell function, with increased cell loss due to increased apoptotic events, is predicted. Using the AdlacZ vector, cell loss by apoptotic mechanisms appears limited to the first days following coculture with high viral concentrations, and does not appear to influence in vitro or in vivo cell function of the surviving islet cells. PMID- 9202641 TI - Functional and histological differences in autogenous and allogenic vein grafts: two different vasculopathies? AB - The long-term biological characteristics and the functional and morphological changes that occur in fresh allografts are poorly understood. This study tests the hypothesis that the development of intimal hyperplasia and its associated functional changes are accelerated in an allograft compared to an autograft due to the additional immunological stimuli. Common carotid vein bypass grafts were performed in 40 New Zealand White rabbits: 20 received their ipsilateral jugular veins (autologous) and 20 received the fresh contralateral jugular veins from the control rabbit (allogenic). Electron microscopy was performed and intimal and medial dimensions were determined by videoplanimetry at 7, 14, and 28 days. Contraction and relaxation studies to a panel of agonists were also performed. The EC50's (agonist concentration which produces 50% of the maximal response) were calculated. All grafts remained patent. Allografts showed a 51% decrease in overall mean intimal thickness (41 +/- 3 microns vs. 83 +/- 12 microns; P < 0.01) and a 97% increase in overall mean medial thickness (140 +/- 15 microns vs. 71 +/ 3 microns; P < 0.01) compared to the autografts. The lumen of the allogenic vein grafts was equivalent to the autologous vein grafts. Overall mean total wall thickness only increased by 17%, 181 microns vs. 154 microns for allo- and autografts, respectively. The EC50 for norepinephrine, histamine, and bradykinin were similar in the auto- and allografts, while the EC50 to serotonin was significantly less in the allografts than in the autografts. Neither the precontracted auto- or allografts relaxed to acetylcholine or serotonin (receptor mediated, endothelium dependent). The EC50 for calcium ionophore (nonreceptor mediated, endothelium dependent) was equivalent in the auto- and allografts. The EC50 for the sodium nitroprusside-induced relaxation (endothelium independent) was significantly higher in the allograft than in the autograft. This study demonstrates that there are two different vasculopathies occurring in autografts and allografts: intimal hyperplasia is predominant in the autograft while an exaggerated medial response is predominant in the allograft. Serotonin contractility and endothelial-independent relaxation are enhanced in the allograft compared to the autograft. PMID- 9202643 TI - Hepatic glutaminase gene expression in the tumor-bearing rat. AB - Previous studies have documented an increase in hepatic plasma membrane glutamine transport in the tumor-bearing rat, but the effects of tumor burden on hepatic glutaminase expression have not been carefully studied. The purpose of this study was to examine the effects of tumor burden and food intake on hepatic glutaminase expression. Rats were implanted with syngeneic methylcholanthrene-induced fibrosarcoma tumor tissue; control rats were sham operated and pair-fed every 24 hr. Northern blotting was used to assay the effect of tumor burden and fasting on hepatic glutaminase mRNA levels, using beta-actin mRNA as a control. Hepatic glutaminase mRNA levels in livers of pair-fed controls were found to be 4-fold greater than levels in livers of tumor-bearing animals. Examination of food intake patterns in these animals indicated that pair-fed controls ate their allotted chow quickly while tumor-bearing rats ate small amounts throughout each 24 hr period. This observation suggested that the differences in glutaminase mRNA levels may be due to a period of fasting by pair-fed animals which was not experienced by the tumor-bearing group. Hepatic glutaminase mRNA levels rapidly increased in normal rats during acute fasting to levels 5.5-fold greater than fed animals. Glucose feeding and insulin injection rapidly reversed the effect of fasting on hepatic glutaminase mRNA levels in normal rats. Tumor-bearing rats also exhibited upregulation of hepatic glutaminase mRNA levels in response to fasting. CONCLUSIONS: (1) Tumor burden itself does not alter hepatic glutaminase expression, at least at the pre-translational level. Instead, differences in hepatic glutaminase mRNA content are due to differences in food intake patterns. (2) Hepatic glutaminase mRNA levels are rapidly upregulated in response to fasting, an effect which appears to be linked to a decrease in plasma insulin concentrations. Because tumor-bearing rats eat regularly over a 24 hr period (albeit in small increments), thereby maintaining the plasma insulin concentration, hepatic glutaminase mRNA may not rise as it does in pair-fed controls whose daily chow intake is complete within hours of food allocation. (3) This study indicates that differences in the timing of food intake between tumor bearing rats and pair-fed controls can alter the expression of genes that are influenced by nutrient availability. These differences should be taken into account when designing studies which involve pair-feeding to control nutrient intake. PMID- 9202644 TI - Decreased myogenic reactivity in skeletal muscle arterioles after hypothermic cardiopulmonary bypass. AB - Cardiopulmonary bypass (CPB) is associated with a generalized defect in the intrinsic control of vascular smooth muscle. To determine if myogenic reactivity of skeletal muscle arterioles was altered by CPB, sheep (n = 7) were placed on hypothermic CPB (27 degrees C) for 90 min and hearts were arrested by cold blood cardioplegia ([K+] = 25 mM) for 60 min. Arterioles (70-180 microns) were isolated from the gracilis muscle before (control) and 15 min after CPB. In vitro arteriolar responses were studied with video-microscopy. Myogenic reactivity was examined to stepwise increases in intraluminal pressure from 10 to 100 mm Hg. Mean arterial pressure was decreased from 80 +/- 15 prior to CPB to 55 +/- 4 mm Hg (P < 0.01) 15 min after CPB. Myogenic contraction was observed in control vessels and was markedly attenuated by the protein kinase C inhibitor staurosporine (P < 0.01). CPB decreased myogenic contraction and shifted the pressure-diameter relation upward, suggesting a decrease in the intrinsic tone (both P < 0.05 vs control). CPB reduced contractile responses to the alpha 1 adrenoceptor agonist phenylephrine from -43 +/- 7% to -23 +/- 5% (P < 0.01) and the protein kinase C activator 12-deoxyphorbol 13-isobutyrate 20-acetate (phorbol ester) from -64 +/- 6% to -38 +/- 16% (P < 0.01). CPB-associated decrease in myogenic reactivity of skeletal muscle arterioles is likely due to alterations in protein kinase C and/or downstream signal transduction. This may account in part for reduction in systemic vascular resistance and hypotension associated with CPB. PMID- 9202645 TI - Retroviral mediated gene transduction alters integrin expression on vascular endothelial cells. AB - Genetically recombinant endothelial cells (rEC) may improve the patency of small diameter vascular grafts by preventing thrombosis or limiting neointimal hyperplasia. Previous work has shown that rEC have reduced adhesion to vascular bypass grafts in vivo. Poor adhesion may be due to altered adhesion (integrin) receptors. This study evaluated the expression of the alpha 5 beta 1 (fibronectin), alpha 2 beta 1 (collagen IV), and alpha v beta 3 (vitronectin) integrin subunits on rEC. Human umbilical vein EC or canine jugular vein EC were transduced with neoR, neoR and human tPA or hygromycin resistance genes using retroviral vectors. Naive EC and EC exposed to empty viral particles (mEC) were controls. Naive EC, mEC, and all rEC's were evaluated for alpha and beta subunits for each integrin receptor studied using immunoblotting. Blotting for alpha 2, alpha 5, and alpha v exhibited expression of the alpha integrin subunits in all cells. The beta 1 and beta 3 subunits were present in mEC and nEC but were absent or truncated in all rEC. The decreased adhesion of rEC's to synthetic vascular grafts may be accounted for by their altered beta 1 and beta 3 integrin subunit expression. The beta subunit is critical for organization of the cytoskeleton and cellular signal transduction. Diminished beta subunit expression in rEC is neither vector specific nor related to retroviral exposure alone. Alteration of beta integrin expression may be to associated with the over-expression of phosphotransferase genes such as neoR or hygromycin B used as selectable markers in gene transfer protocols. PMID- 9202646 TI - Reducing the deleterious effects of intrauterine CO2 during fetoscopic surgery. AB - The fetoscopic approach to fetal intervention is a promising minimally invasive technique for correcting congenital anomalies in utero. However, expansion of the amniotic cavity with CO2 to visualize the fetus causes fetal hypercarbia and acidosis. We assessed whether maternal hyperventilation during intrauterine CO2 insufflation could attenuate the fetal hypercarbic acidosis. Seven fetal lambs of 105 +/- 2 days (mean +/- SEM) gestation (term = 145 days) were instrumented with a carotid arterial catheter in utero. After 7 +/- 1 days of recovery, fetoscopic exposure was obtained with intrauterine insufflation of CO2 at 10 mmHg of intraamniotic pressure. After 30 min, the ewe was hyperventilated at a mean respiratory rate of 23/min for 30 min under continuous insufflation. The uterus was then deflated and following 1 hr of stabilization, and the same protocol of CO2 pneumometrium was repeated. Fetal and maternal arterial blood was sampled at baseline and at 15 min intervals. Fetal PaCO2 increased during 30 min of CO2 insufflation (50.8 +/- 2.8 vs. 72.3 +/- 5.0 mmHg, P < 0.01); however, this change was reversed (to 51.5 +/- 3.0 mmHg, P < 0.01) by 30 min of maternal hyperventilation. The fetus developed acidosis after 30 min of CO2 pneumometrium (pH 7.350 +/- 0.012 vs. 7.236 +/- 0.026, P < 0.01); this was also reversed (to 7.366 +/- 0.019, P < 0.01) by maternal hyperventilation. These results were reproducible during the second CO2 insufflation challenge. Fetal hypercarbic acidosis during fetoscopy with CO2 insufflation is reduced by maternal hyperventilation. PMID- 9202647 TI - Cucurbitacin E targets proliferating endothelia. AB - Tumor vasculature offers a target for drugs directed against proliferating endothelia. We hypothesized that cucurbitacin E (CuE), an actin-disrupting agent, would preferentially inhibit proliferating vs. quiescent endothelia. Log-phase and confluent ECV304 and HUVEC human endothelial cells were exposed to 10-200 nM CuE for 1-96 hr, and toxicity was determined at 2-6 days by sulforhodamine B assay. Confluent ECV cells were scraped by Q-tip to allow proliferation at the margin and exposed to CuE at LD50, and the actin cytoskeleton was stained by rhodamine-phalloidin. Cell cycle analysis was performed by flow cytometry. Log phase cells were significantly inhibited compared to confluent. LD50's of log phase cells were much less than for confluent cells (12 vs. 170 nM, ECV; 13 vs 76 nM, HUVEC). Persistent growth inhibition required 24-96 hr exposure to LD50. Followed less than 6 hr exposure. CuE induced F-actin to accumulate centrally in clumps in newly proliferating cells; actin appeared normal in quiescent cells. CuE treated endothelial cells were not blocked at cytokinesis. CuE preferentially potently inhibits proliferating human endothelia compared to quiescent cells in vitro. CuE induces depolymerization of F-actin in proliferating cells. At low concentrations, cucurbitacin E may potently inhibit vascular proliferation by disrupting actin. PMID- 9202648 TI - Chronic septicemia alters alpha-adrenergic mechanisms in the coronary circulation. AB - The purpose of the present study was to determine the effect of chronic sepsis on alpha-adrenergic responsiveness in the coronary microcirculation. Male Sprague Dawley rats (n = 6) were made septic by intraperitoneal implantation of a gelatin capsule containing 35 mg sterile rat feces and 1 x 10(8) viable colony-forming units of Escherichia coli (strain Sm 018). Control rats (n = 6) were implanted with capsules containing only sterile feces. Forty-eight hours after surgery, subepicardial coronary arterioles (80-170 mm) were isolated. In vitro arteriolar responses were studied in a pressurized, no-flow state with video-microscopy. Chronic sepsis decreased the contractile responses to the alpha 1-adrenoceptor agonist phenylephrine and the protein kinase C (PKC) activator 12-deoxyphorbol 13 isobutyrate 20-acetate. Relaxation responses to the alpha 2-adrenoceptor agonist clonidine, the endothelium-dependent vasodilator adenosine 5'-diphosphate, and the PKC inhibitor staurosporine were reduced, but the relaxation to the endothelium-independent cyclic GMP-mediated vasodilator sodium nitroprusside was preserved. Relaxation to clonidine was inhibited by endothelial denudation or after blockade of nitric oxide synthase. Chronic sepsis may reduce alpha 2 adrenoceptor-mediated relaxation of coronary microvessels by causing endothelial dysfunction. The alpha 1-adrenergic mechanism is downregulated, possibly due to alterations in the receptor and/or downstream signal transduction via PKC. PMID- 9202649 TI - Factors responsible for peritoneal granulocyte apoptosis during sepsis. AB - Apoptosis (Ao), is a process by which cells undergo a form of non-necrotic cellular suicide, the control of which may have significant impact on host immunoresponsiveness to a septic challenge. The aim of this study was to determine (1) if Ao is evident in granulocytes harvested from the blood or peritoneum of septic animals and to what extent this was associated with cell activation and (2) whether the in vivo administration of the TNF inhibitor (TNFbp) alters this process. To assess the first aim, C3H/HeN male mice were subjected to polymicrobial sepsis [cecal ligation and puncture (CLP)] or sham-CLP (Sham) and sacrificed at 4 or 24 hr following CLP. Blood leukocytes and peritoneal exudate cells were harvested, stained with monoclonal fluorochrome conjugated antibodies to granulocytes (Gr1), the activation marker ICAM-1, and either the cell cycle dye, 4',6-diamidino-2-phenylindole, or TUNEL assay (to assess the %Ao+), was used for two- and three-color flow cytometric analysis. Peritoneal exudate cells exhibited increased %Ao+ cells at both 4 and 24 hr post CLP, while blood leukocytes showed a decrease in %Ao+ only at 24 hr. The increase in Ao in the peritoneum was evident only in the Gr1- cell population at 4 hr but was present in both Gr1+ and Gr1- cells at 24 h. Furthermore, the increase in the %Ao+ cells was associated with an increased % of ICAM-1 positive cells. To the extent that TNF affects the 24 hr induction of Ao in peritoneal exudate cells, mice were treated with either 250 micrograms TNFbp/mouse (s.c.) or vehicle control immediately following CLP. The results indicate that administration of TNFbp markedly decreased Gr1+ but not Gr1- cell Ao. Thus, not only does polymicrobial sepsis induce a marked early rise in phagocyte Ao associated with cell activation, but the increase in peritoneal granulocyte Ao, unlike macrophage Ao, is mediated by TNF and/or an agent released by TNF. PMID- 9202650 TI - A defective EGF-receptor in waved-2 mice attenuates intestinal adaptation. AB - While the pathophysiology of intestinal adaptation following small bowel resection (SBR) is not well understood, there is evidence to suggest an important role for epidermal growth factor (EGF) in this process. In waved-2 mice, a naturally occurring mutation results in reduced EGF receptor protein tyrosine kinase activity. We tested the hypothesis that an intact EGF receptor is essential for adaptation by subjecting this strain of mice to SBR. A 50% proximal SBR or sham operation (bowel transection with reanastomosis only) was performed in waved-2, heterozygous, and wildtype mice. After 3 days, adaptation was characterized in the remnant ileum as changes in DNA and protein content per unit length. Villus height and crypt depth were measured, and crypt cell proliferation rates were determined by the percentage of crypt cells taking up 5 bromodeoxyuridine. Following sham surgery, all mice regained their preoperative weight by the third postoperative day. After SBR, all mice gained weight while the waved-2 mice did not. Ileal DNA and protein significantly increased after SBR in wild-type and heterozygous mice while these parameters were unchanged in the waved-2 mice. Villus height and crypt cell proliferation increased in response to SBR in all groups; however, the changes were less pronounced in the waved-2 mice. Adaptation after SBR is impaired in waved-2 mice. Signal transduction by the EGF receptor is a critical component of this response. These data endorse a crucial role for EGF and its receptor in the pathogenesis of intestinal adaptation. PMID- 9202651 TI - Sepsis impairs anastomotic collagen gene expression and synthesis: a possible role for nitric oxide. AB - Although intra-abdominal sepsis is known to impair colon healing by inhibiting anastomotic collagen synthesis, the effect of systemic sepsis on this process is unknown. Endotoxins and cytokines associated with sepsis induce nitric oxide synthesis both systemically and locally within colonic tissue. We hypothesized that systemic sepsis impairs colonic healing and examined a possible correlation with nitric oxide expression. Male Sprague-Dawley rats received intraperitoneal injections of either saline (sham group) or Escherichia coli endotoxin (lipopolysaccharide 1 mg/100 g body weight) at Times -24 and -12 hr (LPS group). All animals underwent laparotomy and left colonic anastomosis at Time 0. At 24 and 96 hr postlaparotomy rats were sacrificed, the anastomoses excised, and [3H] proline incorporation into protein measured as an index of total new protein synthesis (TNP). Digestion with purified collagenase yielded incorporation into the collagen fraction (CDP). Additional sham and LPS-treated rats were sacrificed at 24, 72, and 120 hr, the anastomoses excised, and nitric oxide synthase activity in the tissue measured by the conversion of [3H]-arginine to [3H]citrulline in an ex vivo culture system. Finally, sham and LPS rats were sacrificed at 120 hr for measurement of colon anastomotic bursting pressure. Systemic sepsis significantly impaired new collagen synthesis in anastomotic tissue at 24 hr compared to control samples (P < 0.02). No difference was noted at 96 hr. TNP synthesis was similar in both groups at 24 or 96 hr. Northern blot analysis confirmed a significant decrease in Type I and Type III collagen mRNA expression at 24 hr in septic rats. Anastomotic bursting pressure was also decreased in the septic group (P < 0.003). Sepsis elevated nitric oxide synthase activity in anastomotic tissue 24 hr postanastomosis, when compared to sham tissue (P < 0.0001). These data suggest that systemic endotoxin induces nitric oxide synthesis at the anastomotic site. The simultaneous dysregulation of collagen gene expression and synthesis with decreased anastomotic strength suggests a possible regulatory role for nitric oxide in gastrointestinal healing. PMID- 9202652 TI - Effects of luminal nutrients and small bowel transplants on congenital indirect hyperbilirubinemia. AB - The Gunn rat is an excellent model of Crigler-Najjar syndrome, type 1. In previous studies we demonstrated that heterotopic 15-20-cm jejunal transplants from Wistar rats lowered serum bilirubin levels by 40%, and the reduction was transient (6 weeks). In contrast, orthotopic transplants decreased bilirubin levels by 60% and the effect persisted throughout the 8-week study. This study was initiated to identify the luminal substance(s) which are responsible for the persistent bilirubin-lowering effect of jejunal transplants. Thirty-one Wistar to Gunn 15-20-cm jejunal transplants were randomized to receive daily Thiry-Vella graft irrigation with 5 ml of normal saline (n = 8); bile salts (cholate + deoxycholate, 40 mg/ml, n = 5; fats (Microlipid, 20 mg/ml, n = 5); proteins (Casec caseinate, 40 mg/ml, n = 5); and sugars (Moducal + Polycose, 40 mg/ml, n = 8). Bilirubin levels were measured spectrophotometrically at weekly intervals. At 4 and 8 weeks, enzyme-induced bilirubin conjugation activity was measured using added known amounts of added bilirubin. Irrigation of the transplants with saline, protein, and sugar resulted in moderate (40%) lowering of serum total and indirect bilirubin levels. Fat was significantly more effective, lowering mean total bilirubin levels from 9.6 +/- 0.4 to 1.6 +/- 0.2 mg/dl at 6 weeks. After this time, bilirubin levels increased slightly. Bile salts were slightly less effective, lowering bilirubin levels at 6 weeks by only 75%. However, this effect persisted and at 8 weeks levels averaged 2.4 +/- 0.2 mg/dl. Conjugating enzyme activity in the transplants increased from 1.4 +/- 0.3 to 2.5 +/- 0.5 mg bilirubin conjugated/mg tissue/hr. Luminal fats and bile salts appear to augment enzyme-induced bilirubin conjugation in heterotopic jejunal transplant recipients. PMID- 9202653 TI - Differential display in primary and metastatic medullary thyroid carcinoma. AB - Despite recent advances in the understanding of the role of the RET proto oncogene in the development of familial and approximately 30% of sporadic medullary thyroid carcinomas (MTC), little is known about other genetic events that modify the course and outcome of the disease. We compared the expression of genes in intrathyroidal MTCs to autologous local lymph node metastases by means of mRNA differential display (DDRT-PCR). This is the first report of differential display using surgical specimens of a primary cancer and its metastases. Total RNA was extracted from tumor tissue of two patients with MTC associated with multiple endocrine neoplasia (MEN 2B) and sporadic MTC, respectively. Following reverse transcription (RT), the products were PCR-amplified and separated on a denaturating polyacrylamide gel. RT-PCR products demonstrating differential expression were reamplified and used as probes for Northern blot analysis. Six fragments for which differential expression was confirmed were cloned and sequenced. Resultant sequences were tested for homology to sequences in public data bases, and two novel MTC-derived fragments (MDF-1, MDF-2) were identified. Sensitivity of the method was confirmed by identification of a sequence encoding the calcitonin precursor flanking peptide which is expressed almost exclusively in MTC and normal thyroid C cells. Overexpression of the ribosomal genes S3a and P0 was found in the metastases. Recent reports suggest that components of the translational apparatus act as regulatory mediators of growth, proliferation, and neoplastic change. The altered expression of ribosomal proteins and gene products encoded by MDF-1 or MDF-2 may play an important role in the progression and metastatic spread of MTC. PMID- 9202654 TI - In vivo endotoxin tolerance: impaired LPS-stimulated TNF release of monocytes from patients with sepsis, but not SIRS. AB - In vitro pretreatment of human monocytes (MO) with low-dose lipopolysaccharide (LPSp) inhibits TNF release in response to subsequent LPSa activation. Septic patients are often indistinguishable from patients with systemic inflammatory response syndrome (SIRS). We hypothesized that in vivo exposure to "septic" stimuli impairs subsequent LPSa-stimulated MO TNF production in vitro. Human peripheral MO were obtained after informed consent from controls or patients with sepsis, SIRS, or posttrauma [ACCP/SCCM definitions]. Cells were plated in vitro, incubated 24 hr, and then stimulated with 0-1000 ng/ml LPSa for 4 hr. Parallel control MO were incubated in vitro with 100 ng/ml LPSp for 24 hr and then stimulated with 1000 ng/ml LPSa for 4 hr. Supernatant TNF (mean U/ml +/- SEM) was measured by bioassay. ANOVA was used to determine statistical significance. In vitro LPSp pretreatment markedly inhibited subsequent LPSa-stimulated TNF release. In vitro LPSa-stimulated TNF release was likewise significantly inhibited with MO from septic patients compared to controls. Inhibition was more profound in septic patients with shock (not shown). No impaired TNF release was seen with MO from SIRS or trauma patients. In conclusion, in vivo preexposure to inflammatory stimuli in septic patients alters monocyte regulation in a manner similar to in vitro endotoxin tolerance. Provocative in vitro monocyte LPS stimulation may distinguish patients with sepsis and SIRS. PMID- 9202655 TI - Acute experimental distal colitis alters colonic transit in rats. AB - Data from humans with active distal colitis suggest that the proximal colon exhibits increased contractile activity and delayed transit, whereas the distal colon shows decreased contractile activity and rapid transit. The present study used the acetic acid rat model of experimental colitis to determine the effect of distal colitis on total and regional colonic transit in vivo and on the in vitro contractility of circular smooth muscle from the proximal and distal colon. Distal colitis was induced in rats by intracolonic administration of 4% acetic acid; sham control rats received saline enemas. Control and colitic rats were studied 2 days postenemas. Total colon transit was determined by calculating the geometric center of distribution of a radiolabeled marker (51Cr) instilled into the proximal colon. Regional transit was assessed by expressing the radioactivity in the cecum, proximal and distal colon, and excreted stool as a percent of total radioactivity. Muscle strips from the proximal and distal colon were stimulated with 100 microM acetylcholine (ACh) and 60 mM KCl and the tension was expressed as kilograms per square centimeter. Distal colitis was characterized by decreased total colon transit, increased retention of marker in the cecum and proximal colon, and decreased retention of marker in the distal colon. In vitro contractility studies revealed that distal colitis increased proximal colon circular smooth muscle contractility and decreased distal colon circular smooth muscle contractility to both ACh and potassium. Distal colitis is associated with regional differences in colonic circular smooth muscle contractility, which may contribute to delayed transit in the proximal colon and rapid transit in the distal colon. PMID- 9202656 TI - Endothelial cell effect on smooth muscle cell collagen synthesis. AB - Extracellular matrix (ECM) constitutes the bulk of mature intimal hyperplastic lesions. To determine if endothelial cells (ECs) inhibit smooth muscle cell (SMC) collagen synthesis, bovine aortic SMCs were cultured on plastic semipermeable membranes either alone or opposite ECs. SMCs were pulsed with 4 microCi/ml [3H]proline for 3 days and collagen synthesis was measured as trichloroacetic acid-precipitable counts in conditioned media and in cell lysate fractions. Each was standardized to SMC DNA (cpm/microgram DNA). EC effect on SMC gene expression of type I collagen was studied using Northern analysis. To determine whether TGF beta 1 activation is involved in collagen synthesis regulation, parallel studies were performed in the presence of a neutralizing antibody to TGF-beta 1 (25 micrograms/ml) or aprotinin (a plasmin inhibitor, 200 micrograms/ml). SMCs cocultured with ECs were either untreated or treated with 5 ng/ml TGF-beta 1. The effect of culture substrate on EC-SMC interaction was studied by repeating experiments on type I collagen (CI), matrigel (MG), fibronectin (FN), and type IV collagen (CIV). RESULTS: Compared to SMCs cultured alone, ECs inhibited SMC collagen synthesis by 40 +/- 5% (P < 0.01) and gene expression of type I collagen by 60 +/- 4% (P < 0.01). The addition of neutralizing TGF-beta 1 Ab or aprotinin to SMCs cultured alone had no effect on collagen synthesis. Collagen synthesis in SMCs cultured alone on MG was reduced by 49% and by 16% on CIV when compared to SMCs cultured on plastic (plastic: 100 +/- 12% vs MG: 51 +/- 15% vs CIV: 84 +/- 18%, P = 0.10). Collagen synthesis was increased in SMCs cultured alone on FN (195 +/- 14% vs plastic: 100 +/- 12, P < 0.001) and CI (207 +/- 17 vs plastic: 100 +/- 12, P < 0.001). ECs decreased SMC collagen synthesis by 18 +/- 4% when cocultured on CI (P < 0.05) and by 22 +/- 2% when cocultured on MG (P < 0.05). FN prevented EC inhibition of SMC collagen synthesis. CONCLUSIONS: ECs inhibit SMC collagen synthesis and downregulate SMC type I collagen gene expression. TGF-beta 1 does not appear to be involved in this process. Culture substrate composition can affect SMC collagen synthesis and EC modulation of this process. PMID- 9202657 TI - mRNA differential display of colonic mucosa cells in ulcerative colitis. AB - Involvement of mucosal cells in inflammatory bowel disease (IBD) may be a sequenced process and the molecular difference between involved and uninvolved cells might implicate a possible mechanism in the disease process. The aim of this study was to compare gene expression between involved and uninvolved colonic mucosa cells in an individual with ulcerative colitis (UC) and to clone, sequence, and identify those differentially expressed genes, mRNA differential display was used to identify the gene expression in the mucosa of the UC patient. Anchored oligo(dT) primers and random 5' oligonucleotide 10-mer were used to carry out polymerase chain reaction on reverse-transcribed RNA (RT-PCR). The amplified cDNAs were displayed on a standard sequencing gel and comparisons were drawn between each two lanes representing either involved or uninvolved cells from a specific combination of two primers. Wherever differences were noted between lanes, the bands were reamplified using PCR, cloned into specialized vectors for positive selection, and then confirmed by dot blot. The cloned genes were then sequenced and compared with the GenBank database. About 1200 mRNA species were displayed in the sequencing gel. Among them, 106 fragments were differentially expressed between the two groups. Twenty-five of those differentially displayed gene fragments have been isolated, reamplified, and cloned for sequencing and dot blot analysis. Seventeen of the fragments were differentially expressed using the dot blot technique. Among those 25 gene fragments, 14 have homology to known genes and 11 have no match to any reported genes. Those matched known genes included genes for parathyroid tumor, T cell receptor-beta, alpha-nascent polypeptide-associated complex, ovarian cancer, and myeloblast. This is the first study using mRNA differential display to observe differential gene expression between involved and uninvolved mucosa cells in UC and it shows that differential display is a rapid method for characterizing gene changes in vivo in ulcerative colitis. The results from this study may provide useful information and facilitate further gene studies in this disease. PMID- 9202658 TI - Oral monoterpene therapy (perillyl alcohol) reduces vein graft intimal hyperplasia. AB - The development of intimal hyperplasia is recognized as a major impediment to graft patency. D-Limonenes are monoterpenes with a recognized cytostatic effect on cell proliferation by inhibiting posttranslational isoprenylation of p21ras and other small G-proteins. This study examines the effect of perillyl alcohol, an oral hydroxylated D-limonene, on the development of intimal hyperplasia and its associated smooth muscle cell physiological responses in an experimental model of vein bypass grafting. Twenty New Zealand white rabbits had a right carotid interposition bypass graft using the ipsilateral external jugular vein. Ten animals received chronic oral therapy with a perillyl alcohol (200 mg/kg/day; begun 5 days before surgery and continued until harvest) and 10 control animals received vehicle only. All animals were sacrificed on the 28th postoperative day. Vein grafts were harvested either for morphology/videomorphometry (n = 6 per group) or for in vitro isometric tension studies (n = 4; four 5-mm rings per graft). The cell proliferation and incorporation of [3H]thymidine into the cellular DNA of serum-stimulated rabbit aortic smooth muscle cells was also assessed in the presence of increasing concentrations of perillyl alcohol (10(-9) 10(-4) M). Perillyl alcohol treated vein grafts showed a 22% reduction in overall mean intimal thickness (54 +/- 4 microns vs 69 +/- 3 microns; P = 0.006) but a 25% increase in overall mean medial thickness (86 +/- 4 microns vs 61 +/- 3 microns). The intimal ratio of the perillyl alcohol treated vein grafts decreased by 27% compared to controls. Perillyl alcohol induced norepinephrine and serotonin hypersensitivity in vein grafts compared to controls. The IC50 for perillyl alcohol was 176 nM with maximal inhibition at 5 microM. Incubation of smooth muscle cell cultures with increasing concentrations of perillyl alcohol showed a dose-dependent decrease in in vitro cellular proliferation, maximal at 1 microM. Therapy with perillyl alcohol alters the early development of intimal hyperplasia reducing the intimal response but increasing the medial response without significant changes in the physiological responses of the smooth muscle cells. Modulating G-proteins will affect the intimal hyperplastic response in vein grafts. PMID- 9202659 TI - Cyclic strain is a weak inducer of prostacyclin synthase expression in bovine aortic endothelial cells. AB - Recent studies indicate that hemodynamic forces such as cyclic strain and shear stress can increase prostacyclin (PGI2) secretion by endothelial cells (EC) but the effect of these forces on prostacyclin synthase (PGIS) gene expression remains unclear and is the focus of this study. Bovine aortic EC were seeded onto type I collagen coated flexible membranes and grown to confluence. The membranes and attached EC were subjected to 10% average strain at 60 cpm (0.5 sec deformation alternating with 0.5 sec relaxation) for up to 5 days. PGIS gene expression was determined by Northern blot analysis and protein level by Western blot analysis. The effect of cyclic strain on the PGIS promoter was determined by the transfection of a 1-kb human PGIS gene promoter construct coupled to a luciferase reporter gene into EC, followed by determination of luciferase activity. PGIS gene expression increased 1.7-fold in EC subjected to cyclic strain for 24 hr. Likewise, EC transfected with a pGL3B-PGIS (-1070/-10) construct showed an approximate 1.3-fold elevation in luciferase activity in EC subjected to cyclic strain for 3, 4, 8, and 12 hr. The weak stimulation of PGIS gene expression by cyclic strain was reflected in an inability to detect alterations in PGIS protein levels in EC subjected to cyclic strain for as long as 5 days. These data suggest that strain-induced stimulation of PGIS gene expression plays only a minor role in the ability of cyclic strain to stimulate PGI2 release in EC. These findings coupled with our earlier demonstration of a requisite addition of exogenous arachidonate in order to observe strain-induced PGI2 release, implicates a mechanism that more likely involves strain-induced stimulation of PGIS activity. PMID- 9202661 TI - Interleukin-6 upregulates GP96 expression in breast cancer. AB - Interleukin-6 is associated with poor prognosis in breast cancer. Expression of GP96, a glucose regulated stress protein, is related to drug resistance in tumor cells. Interleukin-6 has previously been shown to induce GP96 expression in a murine myeloblastic cell line. BT474 or MDA-MB231 cells were incubated with recombinant Interleukin-6 (100 to 750 U/ml) for 24 hr. To establish a time course for GP96 induction, MDA-MB231 cells were incubated with 250 U/ml recombinant interleukin-6 for 0-48 hr. Following incubation, cells were washed twice in phosphate-buffered saline (PBS) and cell lysates were prepared by adding 100 microliters of PBS and freezing at -20 degrees C. GP96 was assessed by immunoblotting. Breast tumor tissue and histologically normal breast tissue were obtained within 1 hr of resection and flash frozen in liquid nitrogen. Tissue was homogenized in ice-cold PBS and cell debris was pelleted by centrifugation at 300g at 4 degrees C for 5 min. Supernatants were collected and assayed for interleukin-6 by ELISA, and GP96 by immunoblotting. Both interleukin-6 (P < 0.001) and GP96 are elevated in breast tumor tissue compared to histologically normal tissue. Interleukin-6 (> or = 250 U/ml for > or = 12 hr) induces GP96 in the metastatic breast cancer cell line, MDA-MB231, but has no effect on GP96 levels in the primary cell line, BT474. Elevated interleukin-6 in breast tumors may induce GP96 expression in tumor cells conferring a survival advantage by rendering them resistant to cytotoxic therapy and other forms of stress. PMID- 9202660 TI - IL-6 production in human intestinal epithelial cells following stimulation with IL-1 beta is associated with activation of the transcription factor NF-kappa B. AB - Recent studies suggest that interleukin-1 beta (IL-1 beta) stimulates interleukin 6 (IL-6) production in human intestinal epithelial cells, but the intracellular mechanisms of this response are not known. In other reports, the nuclear factor kappa B (NF-kappa B) regulated IL-6 production in certain cell types. We tested the hypothesis that IL-6 production in the enterocyte is associated with activation of NF-kappa B. Caco-2 cells, a human intestinal epithelial cell line, were grown in tissue culture whereafter they were treated with IL-1 beta (0.5 ng/ml). Cells were preincubated with pyrrolidine dithiocarbamate (PDTC; 10-500 microM), tosyl-lys-chloromethylketone (TLCK; 10-500 microM), or genistein (25-75 microM), all of which are known inhibitors of NF-kappa B. IL-6 levels in the culture media were measured after 24 hr by enzyme-linked immunosorbent assay (ELISA) and IL-6 messenger RNA (mRNA) levels were determined after 4 hr by competitive reverse-transcriptase polymerase chain reaction (RT-PCR). NF-kappa B activity was determined by electrophoretic gel mobility shift assay (EMSA). PDTC, TLCK, and genistein each inhibited IL-1 beta-induced IL-6 production by the Caco 2 cells in a dose-dependent fashion. These responses were also associated with a decrease in IL-6 mRNA levels. There was no NF-kappa B activity in untreated cells, but the addition of IL-1 beta resulted in the activation of NF-kappa B as determined by EMSA. The results suggest that IL-1 beta-induced IL-6 production in the enterocyte is associated with activation of NF-kappa B. The inhibition of IL 6 production by the NF-kappa B inhibitors indicates that the IL-6 production is regulated by NF-kappa B, although further experiments are needed to test that hypothesis. PMID- 9202662 TI - Human growth hormone induces system B transport in short bowel syndrome. AB - BACKGROUND: After massive enterectomy, remnant intestine undergoes compensatory adaptation. A combination of human growth hormone (hGH) and a glutamine-enriched modified diet induces further adaptation in patients with short bowel syndrome (SBS) on long-term total parenteral nutrition. The specific actions of each component, however, are not well-defined. METHODS: New Zealand White rabbits were randomized to control, sham operation, or SBS (70% midjejunoileal resection) groups and treated with either hGH or saline. Sodium-dependent uptake of glucose, glutamine, alanine, leucine, and arginine into brush border membrane vesicles was quantitated. Serum insulin-like growth factor-I (IGF-I) levels were determined by immunoradiometric assay. Mucosal mRNA expression of IGF-I and IGF binding protein 4 (IGFBP-4) was evaluated by northern blot analysis using rat cDNA probes. RESULTS: Glutamine and leucine transports were 33 and 39% greater, respectively, in the hGH-treated versus saline-treated SBS group (P < 0.05), supporting induction of system B amino acid transport. This upregulation was due, in part, to an 88% increase in glutamine carrier capacity (Vmax) with no change in carrier affinity (Km). Both hGH treatment and SBS increased serum IGF-I levels without direct correlation with increased nutrient transport. IGFBP-4 mRNA expression in small bowel mucosa of saline-treated SBS animals was significantly greater than saline-treated unoperated control values. Mucosal IGFBP-4 mRNA was not significantly altered from control in the other study groups. IGF-I mRNA expression was not detected in mucosa, but weak hybridization was noted in rabbit liver. CONCLUSIONS: Human growth hormone accelerates early adaptation in SBS by upregulation of system B carrier capacity. Serum IGF-I levels and mucosal IGF-I and IGFBP-4 mRNA expression did not directly correlate with this enhanced nutrient transport, suggesting that hGH might exert its adaptive effects by mechanisms that are independent from the IGF system in this model. PMID- 9202663 TI - Administration of ATP-MgCl2 after trauma-hemorrhage and resuscitation restores the depressed cardiac performance. AB - Although adenosine triphosphate (ATP)-MgCl2 has been shown to improve cardiac performance under normal and postischemic conditions, it is not known whether this agent has any salutary effects on cardiac performance following trauma hemorrhage and crystalloid resuscitation. To determine this, rats underwent laparotomy (i.e., trauma induction) and were bled to and maintained at a mean arterial pressure of 40 mm Hg until 40% of the maximum shed blood volume was returned in the form of Ringer's lactate. The animals were then resuscitated with four times the volume of shed blood using Ringer's lactate over 60 min and received either ATP-MgCl2 (50 mumole/kg body wt) in 1 ml volume or an equivalent volume of normal saline intravenously over 95 min. Maximum dP/dt during contraction as well as relaxation (+/-dP/dtmax) and ventricular peak systolic pressure (VPSP) were determined 15 min prior to the end of resuscitation and every 30 min thereafter for 4 hr after the completion of resuscitation. The results indicate that both -dP/dtmax and +dP/dtmax decreased significantly beginning at 0 and 2 hr after the completion of resuscitation, respectively, and remained depressed throughout the duration of the study in saline-treated animals. In addition, VPSP was significantly depressed at 2-4 hr after resuscitation. Treatment with ATP-MgCl2, however, restored these parameters. Moreover, the depressed heart rate was also restored following ATP-MgCl2 administration. Since ATP-MgCl2 restores various left ventricular performance parameters, this agent appears to be a promising adjunct for improving cardiac function after trauma and hemorrhage, even in the absence of blood resuscitation. PMID- 9202664 TI - Growth hormone improves immune function and survival in burned mice infected with herpes simplex virus type 1. AB - Recombinant human growth hormone (rhGH) is often used clinically for the treatment of burned patients to promote wound healing and to improve protein metabolism. Recently, the immunomodulatory activity of GH has been described, but it remains unclear whether immune abnormalities associated with burn are improved by rhGH administration. To determine the immunoregulatory activities of rhGH in thermal injury, the survival of burned mice infected with herpes simplex virus type 1 (HSV-1), along with interferon-gamma (IFN-gamma) production by splenic mononuclear cells (SMNCs), and generation of cytostatic macrophages in burned mice were examined after exogenous administration of rhGH. Data showed that the mortality after HSV-1 infection was improved in burned mice treated with a 4 mg/kg dose of rhGH every other day for five doses. Also, the decreased IFN-gamma response of SMNCs from burned mice improved with rhGH therapy. Further, cytostatic macrophages were generated in burned mice treated with rhGH, whereas these macrophages were not demonstrated in burned mice treated with saline. These results indicate that rhGH can improve immune function in burned mice, specifically improving survival in HSV-1 infection, boltering IFN-gamma production by SMNCs, and increasing production of cytostatic macrophages. PMID- 9202665 TI - The effect of epidermal growth factor on the septic complications of acute pancreatitis. AB - Bacterial translocation (BT) from the gastrointestinal tract to mesenteric lymph nodes (MLN) and other extraintestinal organs is an important source of infection in acute pancreatitis (AP). Epidermal growth factor (EGF), a peptide hormone with trophic effects on gut mucosa, has decreased intestinal mucosal injury in septic rats and decreased burn-induced BT in mice. The purpose of this study is to examine whether EGF could affect BT in acute necrotizing pancreatitis. Forty eight male Sprague-Dawley rats (250-350 g) were studied. AP was induced in Group I and Group II by pressure injection of 3% taurocholate and trypsin into the biliopancreatic duct (1 ml/kg of body weight). Group III and Group IV underwent laparotomy without induction of acute pancreatitis. Group I rats received human recombinant EGF (100 micrograms/kg, subcutaneously twice daily) and Group II rats received a similar volume of 0.1% bovine serum albumin as a placebo postoperatively. Group III and Group IV received EGF and placebo, respectively. At 48 hr postoperatively, blood was drawn for culture and amylase determinations. Jejunum and ileum were obtained to measure mucosal protein content, mucosal thickness, villus height, and crypt depth. Specimens from MLN, spleen, liver, pancreas, and cecum were harvested for pathology and culture of gram positive (G+), gram negative (G-), and anaerobic bacteria. Ileal mucosal protein levels were increased significantly in Group I (1.96 +/- 0.14 mg/cm) compared to Group II (0.95 +/- 0.15 mg/cm intestinal segment) (P < 0.01). Jejunal and ileal mucosal thickness, villus height, and crypt depth in Group I were significantly increased when compared to Group II (P < 0.05). All 12 rats in Group II had BT to MLN compared to 58% (7 of 12 rats) in Group I (P < 0.05). Thirty-three percent (4 of 12 rats) had BT to distant sites such as pancreas, spleen, liver, and/or blood in Group I vs 83% (10 of 12 rats) in Group II (P < 0.05). EGF treatment minimizes intestinal damage, decreases BT to MLN and bacterial spread to distant sites, and may be beneficial in preventing septic complications in AP. PMID- 9202666 TI - An in vitro model to assess mucosal immune function and bacterial translocation. AB - The importance of secretory immunoglobulin A (IgA) on intestinal barrier function has gained increasing acceptance. However, due to the complexity of the intestinal microenvironment, the relative role of secretory IgA (sIgA) in mucosal defense has been difficult to study in vivo. Polarized Madin-Darby canine kidney (MDCK) epithelial cells expressing the complementary DNA (cDNA) for the polymeric Ig receptor were grown as monolayers in an in vitro two-chamber cell culture system to study the impact of sIgA on bacterial translocation (BT). Polymeric sIgA or media alone was added to the apical chambers of cell monolayers followed by apical inoculation with bacteria. The basal compartment was sampled at timed intervals thereafter to determine BT. Bacterial passage across the MDCK epithelial cell monolayers occurred in a time and bacterial inoculum concentration gradient. Addition of sIgA led to significant reductions in BT across the epithelial cell monolayers. This is a useful model for further investigation on the role of sIgA in intestinal barrier function. PMID- 9202667 TI - Protein kinase C isoform diversity in preconditioning. AB - Protein kinase C (PKC) appears to be a common intracellular effector and signal collector during cardiac preconditioning; however, it remains unknown whether agonists that activate different PKC isoforms are also linked to select aspects of myocardial protection. Using agonists that are known to activate unique combinations of PKC isoforms, we interrogated the relationship between isoform activation and the different aspects (pH, function, and viability) of endogenous myocardial protection. To study this, isolated rat hearts were subjected to ischemia-reperfusion (I/R) (20 min/40 min), without (control = Ctrl) or with receptor-dependent [phenylephrine (PE), 50 microM; adenosine (ADO), 125 microM] or -independent [phorbol myristate acetate (PMA), 100 nM] activation of PKC. Function, pH, and viability were assessed by rate pressure product (%RPP) and coronary flow (CF; ml/min), by 31P NMR, and by CF creatine kinase (CK; U/liter) leak, respectively. PMA, which activates PKC delta but not eta, resulted in intracellular pH (pHi) and viability protection, but did not protect against postischemic myocardial stunning. ADO, which activates PKC eta but not delta, protects against stunning, but not acidosis or necrosis. PE, which activates PKC delta and eta, provided global myocardial protection against necrosis, acidosis, and stunning. Different PKC isoforms may be linked to distinct aspects of myocardial protection. Targeted activation of PKC isoforms may allow precise mechanistic application of preconditioning-like myocardial protection. PMID- 9202668 TI - Quantitation of in vivo gene delivery by restriction enzyme PCR generated polymorphism. AB - The Multiple intestinal neoplasia (Min) mouse develops multiple polyps in the intestine, due to a heterozygous mutation of the Apc locus. Our laboratory has been introducing normal human adenomatous polyposis coli (APC) gene into the Min mouse through liposome enema to prevent or reverse polyp formation. We have quantitated the amount of normal human APC gene delivered in vivo by a restriction enzyme site specific quantitative PCR. Adult Min and BALB/C mice were treated with lipofectant and human APC complementary DNA (cDNA) plasmid. Min colonic DNA was amplified using primers for Apc nucleotide 2524F (5'2524 TCTCGTTCTGAGAAAGACAGAAGCT) and 2679R (5"2679-TGATACTTCTTCCAAAGCTTTGGCTAT). Highlighted primer sequences were purposely different so as to generate two HindIII restriction enzyme sites in the presence of normal mouse Apc (Apc+). Genomic DNA from untreated Min colonic epithelium revealed two bands: 144 bp for ApcMin and 123 bp for Apc+. BALB/C DNA was amplified using primers flanking a region within the APC gene containing a HindIII site on the human APC, which is absent in the murine APC (Apc). Min's DNA extracted 24 hr after treatment demonstrated a plasmid content of 3% due to a relative increase in the Apc+ (123 bp) band. Six weeks of treatments increased delivery to 10%. APC gene therapy of colonic epithelium can be quantitatively measured through restriction enzyme quantitative PCR. Long-term treatment further increases gene delivery. PCR generated polymorphism is a reliable and reproducible technique to initially optimize transfection conditions and ultimately quantitate efficacy in an in vivo gene delivery model. PMID- 9202669 TI - Construction of recombinant adeno-associated virus vector containing the rat preproinsulin II gene. AB - We have investigated a possible delivery system for the rat preproinsulin II gene (rI2) utilising a recombinant adeno-associated virus (rAAV) vector system, with the long-term goal of engineering stably infected insulin-producing cell lines. The rAAV vector was chosen because it is a safe and nonpathogenic method for gene transfer. The plasmid pBC12BI (ATCC) was purified and digested with restriction enzymes SepI and StuI to release a fragment containing the Rous sarcoma virus long terminal repeat (RSV-LTR) promoter-driven rat preproinsulin II gene (rI2). Subsequently, the RSV-rI2 gene fragment was cloned into the BamHI site of rAAV vector plasmid pWP-19 to produce the rI2 recombinant plasmid designated pLP-1. The pWP-19 also encodes the AAV inverted terminal repeats for integration and replication and the herpes virus thymidine kinase promoter-driven gene for neomycin resistance (neoR). The cell line 293 (ATCC) was then cotransfected with pLP-1 and helper plasmid pAAV/AD, which is required for viral replication. The rAAV genome, now containing rI2, was rescued using adenovirus and packaged into mature AAV virions termed vLP-1. Finally, human pancreatic adenocarcinoma cells (HPAC; ATCC) were exposed to vLP-1, selected for G418 resistance, and screened for insulin production. Successful rescue was confirmed by Southern blot analysis using the rI2 gene probe derived from the original plasmid. The final titer of 1.25 x 10(9) particles/ ml was determined by DNA slot blots using pLP-1 as the standard, HPAC cells were infected with vLP-1 (termed HPAC/rI2). Integration of the rI2 genome in G418-resistant clones was confirmed by Southern blot analysis and again after 6 months in culture by amplification of the rI2 gene by PCR. Insulin gene transcription was confirmed by RT-PCR. We have developed a rAAV mediated gene transfer system for the rat preproinsulin II gene. Successful transduction and stable integration of rI2 into HPAC was achieved. Production of insulin by HPAC/rI2 was confirmed by RIA and RT-PCR, validating this system as an effective approach to experimental gene therapy. PMID- 9202670 TI - Activation of Raf-1 in human pancreatic adenocarcinoma. AB - Point mutations in the Ras oncogene cause Ras to remain in its active GTP-bound state sending signals downstream continuously. Since 75 to 90% of all human pancreatic ductal adenocarcinomas harbor activating mutations at codon 12 of the K-ras oncogene it was our belief that Raf-1-MEK-MAPK will be activated in the majority of human pancreatic cancers. The aim of this study was to confirm activation of Raf-1 in K-ras mutant human pancreatic cancer. Additionally, we sought to determine if Raf-1 activation differed in K-ras mutant and nonmutant pancreatic cancer. Furthermore, we were interested in determining if Raf-1 activation in pancreatic cancer led to subsequent activation of downstream effectors such as MAP kinase. The presence of mutations in codon 12 of the K-ras oncogene in 14 human pancreatic adenocarcinoma cell lines was determined by use of mutant allele-specific PCR restriction fragment length polymorphism analysis. Raf-1 expression of quiescent cells was determined by immunoblotting using a rabbit anti-human polyclonal antibody and enhanced chemiluminescence. MAP kinase activity was determined by measuring the incorporation of phosphate into Myelin Basic Protein. Seven cell lines were noted to have mutations in codon 12 of K-ras while seven cell lines did not. There was no difference in expression of the 74 kDa-activated form of Raf-1 in K-ras mutant vs K-ras nonmutant cell lines. However, there was a significant increase in MAP kinase activity in the nonmutant cell lines compared to the cell lines with Ras mutations (P = 0.026). We conclude that Raf-1 is expressed in its active form in human pancreatic cancer regardless of K-ras status. However, signalling downstream of Raf-1 differs in cell lines with K-ras mutations compared to those cell lines without K-ras mutations. PMID- 9202672 TI - Protein kinase C inhibits epidermal growth factor receptor phosphorylation in enterocytes. AB - Epidermal growth factor (EGF) is an important proliferative signal in the gastrointestinal tract. The EGF receptor (EGFr), which transduces the mitogenic stimulus to the cell, may be regulated by a number of factors including extracellular matrix, cell-cell contact, and other peptides. As protein kinase C (PK-C) has been shown to phosphorylate and down-regulate the EGFr in certain tumor cell lines, we propose that PK-C, an important regulatory enzyme, modulates the phosphorylation of the EGFr in the IEC 6 rat enterocyte cell line. IEC 6 cells were cultured in dishes with Dulbecco's modified Eagle's medium, (DMEM)/5% fetal bovine serum (FBS), which was changed to DMEM/1% FBS 24 hr prior to all experiments. Cells (three dishes per group) were treated with the PK-C activating phorbol ester phorbol-12-myristate-13-acetate (PMA) (100 nM) or vehicle for 1 hr and challenged with EGF (50 ng/ml) or vehicle for 15 min. Cell lysates were then prepared. EGFr tyrosine phosphorylation was determined by immunoprecipitating the EGFr and immunoblotting with an antibody against phosphotyrosine. EGFr apparent molecular weight was assessed in the same lysates by Western blot with an anti EGFr antibody. Blots were analyzed by computer densitometry. Data are expressed as mean +/- SEM; n = 3 with P value determined by t test. Exposure of cells to PMA resulted in a decrease in the EGF-stimulated EGFr phosphotyrosine content from 96 +/- 5 U in control to 66 +/- 6 U in PMA (P < 0.01). The amount of receptor did not change, 43 +/- 3 U in control vs 44 +/- 3 U in PMA (P = 0.44). Further, exposure to PMA in the absence of EGF caused a gel shift of the EGFr band consistent with a nontyrosine phosphorylation of the protein. We demonstrate that activation of PK-C results in a modification of the EGFr coincident with inhibition of EGF-stimulated receptor tyrosine kinase activity. These data support a role for PK-C in the regulation of EGFr function and hence modulation of mitogenic signals in enterocytes. PMID- 9202671 TI - The contribution of inflammatory mediators and nitric oxide to lipopolysaccharide induced intussusception in mice. AB - Intussusception is a major cause for intestinal obstruction in children. Its etiology is unclear, but it is often associated with some kind of infection. We have developed a model for intussusception in mice using intraperitoneal (IP) injection of lipopolysaccharide (LPS). The objective of this study was to identify the putative mediators that participate in this LPS-induced intussusception. LPS (12 mg/kg) was injected into adult mice (N = 52) and 6 hr later, 25% of the animals demonstrated intussusception in the small or large intestine. We next tested whether nitric oxide (NO) or various inflammatory mediators contributed to this effect: Indomethacin (10 mg/kg) injected with LPS (12 mg/kg) completely prevented the effect of LPS (N = 20). The tumor necrosis factor (TNF) blocker pentoxifylline (200 mg/kg) significantly reduced the incidence of intussusception to 6.6% (N = 30). The platelet-activating factor (PAF) antagonist BN52021 (10 and 20 mg/kg) reduced the incidence of intussusception to 13.3% in both doses (N = 15 for each dose). Addition of 2% arginine (NO precursor) to the drinking water 36 hr before the injection of LPS increased the incidence of intussusception to 30.7% (N = 32). In mice injected with the NO synthase inhibitor L-NAME (20 mg/kg) only 3.8% developed intussusception (N = 26). Our results indicate that the induction of intussusception by LPS proceeds via parallel pathways involving cytokines, prostaglandins, and NO. Our previous pathological study showed that LPS did not cause any changes that may act as a lead point for the intussusception, suggesting that LPS induced intussusception by altering gut motility. We therefore propose that these mediators combine to induce disturbed gut motility that results in the formation of intussusception. PMID- 9202673 TI - Relationship between energetic, ionic, and functional status in the perfused rat heart following thermal injury: a 31P and 23Na NMR study. AB - To test the hypothesis that energy deficits and intracellular ion derangements may be the cellular basis for intrinsic myocardial dysfunction in rats after burn trauma, we examined ATP metabolism, intracellular pH, sodium, and mechanical performance simultaneously in perfused beating hearts from sham burn or burned rats (43% TBSA, 3 degrees scald burn, resuscitated for 24 hr with lactated Ringer's solution, Parkland formula). Intracellular calcium was also measured in myocytes harvested from parallel groups of sham burn and burn resuscitated rats. Burn trauma caused a 46% decrease in left ventricular developed pressure, a 69% decrease in +dP/dtmax, and a 72% decrease in -dP/dtmax. Intracellular to external standard sodium ratio increased (+58%) from 0.318 +/- 0.027 to 0.500 +/- 0.048 (P < 0.05), and intracellular calcium increased (+67%) from 206 +/- 13 to 445 +/- 37 nM (P < 0.01). Burn hearts exhibited decreased functional response to isoproterenol challenge compared to sham burn controls, but energy metabolism was similar in all hearts, regardless of burn injury. Our data suggest that burn trauma alters intracellular cardiomyocyte calcium and sodium homeostasis, and ionic derangements are not related to either altered intracellular pH or high energy phosphate deficits. PMID- 9202674 TI - Modulation of protein tyrosine phosphorylation by the extracellular matrix. AB - Fibronectin (FN) cross-linked to fibrin following injury provides the provisional matrix required for cells to begin tissue repair. Our previous work has demonstrated that fibroblasts adherent to multimeric FN within the context of a fibrin matrix (FN-fibrin) exhibit clear phenotypic differences from those adherent to a dimeric FN-coated surface. We hypothesize that this response to multimeric FN may be mediated by altered protein tyrosine phosphatase activity following integrin activation. METHODS: NIH 3T3 cells were plated in the presence or absence of pervanadate (PV), a phosphotyrosine phosphatase inhibitor, on wells coated with FN or FN-fibrin matrix. Spread cell areas were measured after increasing incubation times and are recorded as mean cell area (mm2) +/- SEM. Alternatively, cells were lysed and equal amounts of protein were analyzed by immunoblot using a monoclonal antibody specific for phosphotyrosine. RESULTS: PV significantly inhibited cell spreading on FN-fibrin matrices. In contrast, PV treatment had little effect on cell area on FN alone. Analysis of cell lysates revealed that protein tyrosine phosphorylation events differ in a substrate dependent manner. CONCLUSION: Cell attachment to a FN-fibrin matrix induces distinct cell shape and cytoskeletal organization. Inactivation of tyrosine specific phosphatases enhances this distinction and inhibits the spreading of cells attached to this substrate. The phosphotyrosyl protein content of treated cells on FN-fibrin matrix is also diminished. These results suggest that cell extracellular matrix interactions affect the tyrosine phosphorylation balance of the cell, thus modifying cytoskeletal organization and related signaling events. PMID- 9202676 TI - Pharyngoesophageal reconstruction with the use of vascular anastomoses: operative modifications and long-term prognosis. AB - OBJECTIVE: Vascular surgical techniques have contributed to the success of pharyngoesophageal reconstruction. We report our methods and analysis of postoperative complications, quality of life, and long-term prognosis. METHODS: Sixty-seven patients who underwent pharyngoesophageal reconstruction with use of vascular anastomoses comprised the study population. The operative procedures performed were free jejunal autograft transplantation in 54 patients, gastric pedicle placement with vascular anastomoses in 2, jejunal pedicle with vascular anastomoses in 4, colonic pedicle with vascular anastomoses in 4, free jejunal graft and gastric pedicle in 2, and free jejunal graft and jejunal pedicle in 1. The common carotid artery and internal jugular vein were primarily used as the recipient vessels. The period of postoperative observation ranged from 3 days to 145 months. RESULTS: The postoperative complications noted were dehiscence in 7 patients, graft failure in 1, wound infection in 2, small bowel intussusception in 4, pneumonia in 2, disseminated intravascular coagulation in 1, and pancytopenia in 1. Revascularization was successful in all but 1 patient, and oral intake was achieved in 58. Persistent swallowing dysfunction was recognized in 4%. Speech restoration was achieved in 57% of the patients with esophageal speech in 7% and with an artificial larynx in 50%. In the long-term follow-up, 36% of our patients died of the primary disease, 9% died of other diseases, and 55% are alive. CONCLUSIONS: Esophageal reconstruction with the use of vascular anastomoses affords low morbidity and mortality. Postoperative swallowing and speech are satisfactory, and the function of the reconstructed esophagus is well preserved for as long as 10 years. PMID- 9202675 TI - In vivo adenoviral-mediated gene transfer in the treatment of pancreatic cancer. AB - Gene therapy may allow targeted delivery of tumoricidal drugs to treat pancreatic cancer. Cytosine deaminase (CD) is a bacterial enzyme that converts the nontoxic agent 5-fluorocytosine (5FC) to the active chemotherapeutic agent 5-fluorouracil (5FU). Neoplastic cells induced to express the CD gene treated with 5FC may generate locally high concentrations of 5FU while minimising systemic toxicity. Replication deficient adenovirus vector carrying the CD gene (AdCMV.CD) was tested for therapeutic efficacy against the murine pancreatic carcinoma cell line Pan02. Pan02 cells were infected in vitro with AdCMV.CD or null vector (Ad.-Null) and were examined for expression of CD messenger RNA (mRNA) (Northern blot) and CD enzymatic function (spectrophotometry). mRNA transcripts of the CD gene increased in a dose-dependent manner after infection with AdCMV.CD. Conversion of 5FC to 5FU at a multiplicity of infection (MOI) of 20 was measured to be 51% after a 48-hr incubation. Growth inhibition was measured by MTT assay and thymidine uptake. Pan02 growth in vitro treated with AdCMV.CD and 5FC was inhibited by 80% as compared to cells treated with Ad.Null and 5FC. An in vivo model of pancreatic cancer was established by injecting 2.5 x 10(5) PAN02 cells subcutaneously into the flanks of C57BL/ 6 mice. Seven days later AdCMV.CD was injected into each tumor and 5FC was administered for 10 days. Treatment of mice with AdCMV.CD and 5FC inhibited tumor growth compared to mice who received AdCMV.CD only or 5FC only. These data demonstrate the therapeutic efficacy of an enzyme prodrug strategy in experimental pancreatic cancer. PMID- 9202677 TI - Diagnostic and therapeutic thoracic surgery in leukemia and severe aplastic anemia. AB - BACKGROUND: Pulmonary complications often occur in patients with leukemia and severe aplastic anemia. Particularly in patients having undergone bone marrow transplantation, respiratory diseases account for a significant number of deaths. In a retrospective study, we evaluated 41 patients with leukemia and severe aplastic anemia who were operated on consecutively from 1980 to 1995. METHODS: Fourteen open lung biopsies, four video-assisted lung biopsies, and 24 lung resections were performed in 24 male and 17 female patients. Mean age was 32.2 years. RESULTS: Eleven (27%) early deaths occurred (30-day mortality): ten in patients after lung biopsy (56%) and one after lung resection (4%) (p < 0.001). Perioperative morbidity relating to pulmonary disease or operation included 10 (24.4%) cases of prolonged (> 24 hours) postoperative mechanical ventilation and two (4.8%) cases of bleeding or hematoma. In one (2.4%) patient a slowly developing, contained bronchial stump insufficiency appeared after lobectomy, which was successfully operated on 3 weeks later. CONCLUSION: We conclude that resection of localized pulmonary lesions, be it for diagnostic or therapeutic (or combined) purposes, can be carried out with low morbidity and mortality in patients with leukemia and severe aplastic anemia. However, early mortality is high after open or thoracoscopic lung biopsies in patients with acute-onset diffuse pulmonary disease, and little therapeutic benefit is realized in these cases. PMID- 9202678 TI - Coagulation factor abnormalities after the Fontan procedure and its modifications. AB - OBJECTIVE: Recently we reported the prevalence of thromboembolism in patients who underwent the Fontan procedure and its modifications. Although hemodynamic factors may well contribute to thromboembolism, recent evidence suggests that coagulation factor abnormalities may also play a role. We therefore set out to investigate the coagulation status in a group of patients who had undergone the Fontan procedure. METHODS: The study population consists of 20 children who had undergone the Fontan procedure and its modifications. They were examined for coagulation factor abnormalities. Concentrations of serum albumin, total protein, and liver enzymes were also measured. The median age at the time of the operation was 6.2 years (17 months to 8 years) with a male/female ratio of 2.3:1. The median time from the Fontan repair was 4.9 years (18 to 76 months). RESULTS: Protein C (p < 0.001), protein S (p < 0.02), and factor VII (p < 0.001) were significantly lower than the normal range. The changes in serum albumin and total protein and factors II, IX, and X were not significant. CONCLUSIONS: It is possible that deficiency in protein C, protein S, and factor VII partly account for the prevalence of thromboembolism after Fontan-type repairs. The risk of long term anticoagulation should be weighed against the best palliative procedure for these patients. We suggest that reduced protein C, protein S, and factor VII levels in this group of patients should be regarded as risk factors and that such patients should be treated with anticoagulants. PMID- 9202679 TI - Myocardial protection in normal and hypoxically stressed neonatal hearts: the superiority of blood versus crystalloid cardioplegia. AB - OBJECTIVES: Blood cardioplegia predominates in the adult because it provides superior myocardial protection, especially in the ischemically stressed heart. However, the superiority of blood over crystalloid cardioplegia in the pediatric population is unproved. Furthermore, because many pediatric hearts undergo a preoperative stress such as hypoxia, it is important to compare the different methods of protection in both normal and hypoxic hearts. METHODS: Twenty neonatal piglets were supported by cardiopulmonary bypass and subjected to 70 minutes of cardioplegic arrest. Of 10 nonhypoxic hearts, five (group 1) were protected with blood cardioplegia and five (group 2) with crystalloid cardioplegia (St. Thomas' Hospital solution). Ten other piglets underwent 60 minutes of ventilator hypoxia (inspired oxygen concentration 8% to 10%) before cardioplegic arrest. Five (group 3) were then protected with blood cardioplegia and the other five (group 4) with crystalloid cardioplegia. Myocardial function was assessed by means of pressure volume loops and expressed as a percentage of control. Coronary vascular resistance was measured with each infusion of cardioplegic solution. RESULTS: No difference was noted between blood (group 1) or crystalloid cardioplegia (group 2) in nonhypoxic hearts regarding systolic function (end-systolic elastance 104% vs 103%), diastolic stiffness (156% vs 159%), preload recruitable stroke work (102% vs 101%), or myocardial tissue edema (78.9% vs 78.9%). Conversely, in hearts subjected to a hypoxic stress, blood cardioplegia (group 3) provided better protection than crystalloid cardioplegia (group 4) by preserving systolic function (end-systolic elastance 106% vs 40%; p < 0.05) and preload recruitable stroke work (103% vs 40%; p < 0.05); reducing diastolic stiffness (153% vs 240%; p < 0.05) and myocardial tissue edema (79.6% vs 80.1%); and preserving vascular function, as evidenced by unaltered coronary vascular resistance (p < 0.05). CONCLUSION: This study demonstrates that (1) blood or crystalloid cardioplegia is cardioprotective in hearts not compromised by preoperative hypoxia and (2) blood cardioplegia is superior to crystalloid cardioplegia in hearts subjected to the preoperative stress of acute hypoxia. PMID- 9202680 TI - Inhaled nitric oxide, right ventricular efficiency, and pulmonary vascular mechanics: selective vasodilation of small pulmonary vessels during hypoxic pulmonary vasoconstriction. AB - OBJECTIVE: In the setting of acute pulmonary artery hypertension, techniques to reduce right ventricular energy requirements may ameliorate cardiac failure and reduce morbidity and mortality. Inhaled nitric oxide, a selective pulmonary vasodilator, may be effective in the treatment of pulmonary artery hypertension, but its effects on cardiopulmonary interactions are poorly understood. METHODS: We therefore developed a model of hypoxic pulmonary vasoconstriction that mimics the clinical syndrome of acute pulmonary hypertension. Inhaled nitric oxide was administered in concentrations of 20, 40, and 80 ppm. RESULTS: During hypoxic pulmonary vasoconstriction, the administration of nitric oxide resulted in a significant improvement in pulmonary vascular mechanics and a reduction in right ventricular afterload. These improvements were a result of selective vasodilation of small pulmonary vessels and more efficient blood flow through the pulmonary vascular bed (improved transpulmonary vascular efficiency). The right ventricular total power output diminished during the inhalation of nitric oxide, indicating a reduction in right ventricular energy requirements. The net result of nitric oxide administration was an increase in right ventricular efficiency. CONCLUSION: These data suggest that nitric oxide may be beneficial to the failing right ventricle by improving pulmonary vascular mechanics and right ventricular efficiency. PMID- 9202681 TI - Mechanisms underlying degeneration of cryopreserved vascular homografts. AB - OBJECTIVE: To analyze the mechanism(s) underlying homograft degeneration, we designed an experimental model in which the behavior of cryopreserved autografts and homografts, as well as fresh autografts, implanted in the same animal was compared. METHODS: A cryopreserved homograft was implanted in the aorta of 14 sheep. The excised aortic autologous segment was then subjected to cryopreservation, and 1 to 8 weeks later it was implanted 1 to 2 cm below the cryopreserved homograft. The intermediate segment of the native aorta, the fresh autograft, was dissected at this point. Animals were put to death at different times and the implanted segments were harvested together with a portion of native aorta. Histologic and immunohistochemical analyses, as well as cell viability assessments, were then performed on the explanted segments. Similar studies were also conducted on fragments of cryopreserved autografts and homografts before implantation. RESULTS: With the exception of a partial loss of the endothelium, cryopreserved specimens retained cell viability and morphologic integrity before implantation. Explanted cryopreserved homografts showed profound changes affecting all strata, as well as a decline in cell viability. Lymphocyte infiltrates were found up to 12 months after implantation. Endothelium was always absent in cryopreserved homografts. However, a reendothelialization of the cryopreserved autografts was observed. After an initial period of neuronal degeneration, reenervation of the cryopreserved autograft segment occurred 6 to 12 months after the operation. Findings regarding the fresh autografts were similar to those of the cryopreserved autografts. CONCLUSION: Our results suggest that the immunologic reaction rather than the cryopreservation process is responsible for the degenerative process occurring in cryopreserved homografts. PMID- 9202682 TI - Minimally invasive mitral valve replacement: port-access technique, feasibility, and myocardial functional preservation. AB - OBJECTIVE: This experiment examined the feasibility of minimally invasive port access mitral valve replacement via a 2.5 cm incision. METHODS: The study evaluated valvular performance and myocardial functional recovery in six mongrel dogs after port-access mitral valve replacement with a St. Jude Medical prosthesis (St. Jude Medical, Inc., St. Paul, Minn.). Femoro-femoral cardiopulmonary bypass and a balloon catheter system for myocardial protection with cardioplegic arrest (Heartport, Inc., Redwood City, Calif.) were used. The mitral valve was replaced through a 2.5 cm port in the left side of the chest, and the animals were weaned from bypass. Cardiac function was measured before and at 30 and 60 minutes after bypass. Left ventricular pressure and electrical conductance volume were used to calculate changes in load-independent indexes of ventricular function. RESULTS: Each procedure was successfully completed. Recovery of left ventricular function was excellent at 30 and 60 minutes after bypass compared with the prebypass values for elastance (30 minutes = 4.04 +/- 0.97 and 60 minutes = 4.27 +/- 0.57 vs prebypass = 4.45 +/- 0.96; p = 0.51) and for preload recruitable stroke work (30 minutes = 76.23 +/- 4.80 and 60 minutes = 71.21 +/- 2.99 vs prebypass = 71.23 +/- 3.75; p = 0.45). Preload recruitable work area remained at 96% and 85% of baseline at 30 and 60 minutes (p = not significant). In addition, transesophageal echocardiography demonstrated normal prosthetic valve function, as well as normal regional and global ventricular wall motion. Autopsy revealed secure annular-sewing apposition and normal leaflet motion. CONCLUSIONS: These results suggest that minimally invasive mitral valve replacement using percutaneous cardiopulmonary bypass with cardioplegic arrest is technically reproducible, achieves normal valve placement, and results in complete cardiac functional recovery. Minimally invasive mitral valve replacement is now feasible, and clinical trials are indicated. PMID- 9202683 TI - Heart reduction surgery: an analysis of the impact on cardiac function. AB - OBJECTIVES: Reports of improved ejection fraction, coupled with decreased filling pressures, have prompted a number of centers to begin evaluating the efficacy of heart reduction surgery to ameliorate symptoms of heart failure. However, the impact of this operation on cardiac mechanics is unknown. We applied a multiple compartment elastance model to simulate the effects of excising cardiac mass on heart function. METHODS: The left ventricle was divided into two functional compartments to simulate excision of part of the wall. At multiple increments of mass reduction, the resulting end-systolic elastance, ejection fraction, stroke volume, end-diastolic pressure and volume, and diastolic stiffness were determined. RESULTS: Changes in systolic function were accompanied by offsetting changes in diastolic function; consequently, overall pump function (the Frank Starling Relationship) was found to be depressed. The geometric rearrangement associated with this operation leads to a reduction in wall stress for a given level of pressure generation, thus implying an increase in the efficiency with which wall stress is transduced into intraventricular pressure. CONCLUSIONS: Overall pump function is depressed in the short run after heart reduction surgery. However, on the basis of theoretic arguments, heart reduction surgery may have long-term beneficial implications. Importantly, this analysis revealed that changes in parameters of ventricular function have different implications during heart reduction surgery than when such changes are observed with inotropism caused by acute pharmacologic therapy. PMID- 9202684 TI - Cardiac transplantation for auxiliary circulatory support. PMID- 9202685 TI - Heterotopic cardiac transplantation in infants and children. AB - BACKGROUND: Children with advanced heart failure, particularly those with elevated pulmonary vascular resistance, pose a difficult management problem because the normal donor right ventricle cannot cope with the high pulmonary resistance and because of the relative shortage of donor organs of an appropriate size for this age group. METHODS: In an attempt to address these issues and evaluate the role of heterotopic transplantation in this context, we operated on 12 children, six boys and six girls, in the period between January 1, 1991, and March 31, 1996. Their ages ranged from 11 months to 15.2 years (mean 81.6 +/- 62.8 months) and their mean weight was 23.3 kg (range 7.6 to 56.8 kg). Eight patients (66.6%) had significant elevation of pulmonary artery pressure (pulmonary artery systolic pressure = 66 +/- 9.4 mm Hg, mean transpulmonary gradient = 22.3 +/- 3.4 mm Hg). In all patients the donor pulmonary artery was anastomosed to the recipient right atrium without the use of any prosthetic material. Ischemic times varied between 135 and 255 minutes (mean 182.1 +/- 30.7 minutes). The immunosuppression regimen included cyclosporine and azathioprine. Steroids were not routinely used. RESULTS: One patient died in the hospital of acute rejection on postoperative day 16. Three patients had lobe collapse within 1 week and all were treated successfully. Two late deaths (18.2%) occurred as a result of cardiac rejection 3 months and 2 years after the operations. Nine survivors (75%) are alive, active, and growing normally at a mean follow-up of 2.2 years (range 11 months to 4.75 years). Repeated cardiac catheterization performed in seven patients with preoperative pulmonary hypertension showed a slow progressive drop in mean pulmonary artery pressure. No significant change was observed in the function of the recipient hearts. CONCLUSION: We conclude that heterotopic heart transplantation is feasible for a selected group of children with good medium-term results, notably regression of pulmonary artery pressure, normal growth, and lack of long-term chest complications. PMID- 9202686 TI - Exogenous surfactant treatment before and after sixteen hours of ischemia in experimental lung transplantation. AB - OBJECTIVE: A syngeneic, acute, double lung transplant model in the rat was used to determine the impact of exogenous surfactant treatment on graft function after prolonged cold storage. METHODS: The donor grafts were flush perfused, preserved for 16 hours, and then reperfused for 120 minutes. Untreated lungs served as controls (group I). In group II the recipient received a 200 mg/kg dose of surfactant (CuroSurf) before reperfusion. In groups III and IV, surfactant was administered before perfusion and harvesting (III, 20 mg/kg; IV, 200 mg/kg). Serial measurements of graft pulmonary vascular resistance, alveolar-arterial oxygen difference, and compliance were obtained. Final graft assessment included weight gain and histologic study. RESULTS: Repeated-measures analysis of variance showed significant improvement of graft performance in respect to compliance, alveolar-arterial oxygen difference, and pulmonary vascular resistance in donor surfactant treatment group IV (200 mg/kg) in comparison with recipient treatment (group II) and untreated controls (group I). Reducing the donor surfactant supplementation from 200 mg/kg to 20 mg/kg (group III) improved oxygenation and lung compliance as compared with untreated controls. Grafts in groups I and II had significantly more weight gain after 2 hours of reperfusion. Recipient treatment resulted in significantly more pulmonary hemorrhage in histologic sections. CONCLUSION: Donor treatment with exogenous surfactant is advantageous for preservation of graft function after extended ischemia. Positive effects may be seen with as little as 20 mg/kg of exogenous surfactant given before donor organ perfusion. PMID- 9202687 TI - Intracoronary gene transfer of immunosuppressive cytokines to cardiac allografts: method and efficacy of adenovirus-mediated transduction. AB - OBJECTIVE: Allograft-targeted immunosuppressive gene therapy may inhibit recipient immune activation and provide an alternative to systemic immunosuppression. We studied the optimal technique and efficacy of intracoronary gene transfer of viral interleukin-10 and human transforming growth factor-beta 1 in a rabbit model of heterotopic heart transplantation. METHODS: Replication defective adenoviral vectors were constructed, expressing viral interleukin-10 (AdSvIL10) or transforming growth factor-beta 1 (AdCMVTGF-beta 1). Intracoronary delivery of vectors was accomplished ex vivo by either bolus injection or slow infusion. The allografts were implanted heterotopically in recipient rabbits and collected 4 days after the operation. Vector dose was 4 x 10(9) to 6 x 10(10) pfu/gm of donor heart. Transfer was confirmed by DNA amplification for both genes. Gene product expression in tissue was quantified by immunoassay and visualized by immunohistochemical staining. RESULTS: Allograft viral uptake was only 9.9% +/- 2.4% with bolus injection, but increased to 80.5% +/- 6.8% at 1 ml/min infusion rate (p = 5 x 10(-14)). Uptake ratio was not affected by vector quantity or slower infusion rates. Transforming growth factor-beta 1 was consistently detected in allografts infected with AdCMVTGF-beta 1, but not with control adenovirus or AdSvIL10. Expression was proportional to infused vector quantity and reached 10 ng/gm of allograft at infused 10(10) pfu/gm. Transforming growth factor-beta 1 was also detected in recipient's serum at less than 1 ng/ml. Viral interleukin-10 was detected in minor amounts only (< 1 ng/gm) in allografts infected with AdvIL10 up to 5 x 10(10) pfu/gm. Nevertheless, it was detected in recipient serum at concentrations up to 0.4 ng/ml. CONCLUSIONS: Intracoronary gene transfer of immunosuppressive cytokines to cardiac allografts during cold preservation is feasible. Slow infusion is superior to bolus injection. In vivo effects on allograft rejection remain to be determined. PMID- 9202688 TI - Aspartate/glutamate-enriched blood does not improve myocardial energy metabolism during ischemia-reperfusion: a 31P magnetic resonance spectroscopic study in isolated pig hearts. AB - OBJECTIVE: Our objective was to test the effects of exogenous L-aspartate and L glutamate on myocardial energy metabolism during ischemia-reperfusion. METHODS: Phosphorus 31-magnetic resonance spectroscopy was used to observe cellular energetics and intracellular pH in isolated pig hearts perfused with blood (group A, n = 8) or blood enriched with 13 mmol/L each of L-aspartate and L-glutamate (group B, n = 6). The hearts were subjected to 30 minutes of total normothermic ischemia and then reperfused for 40 minutes. Two hearts from each group were inotropically stimulated by titration with calcium after normokalemic reperfusion. Left ventricular function was measured with the use of a compliant balloon and oxygen consumption was calculated. RESULTS: Magnetic resonance spectroscopy showed no decrease in the rate of energy decline during ischemia for group B versus group A. No significant differences were observed between the two groups in terms of myocardial function, oxygen consumption, or the rate or extent of high-energy phosphate recovery after normokalemic reperfusion or inotropic stimulation. Inotropic stimulation of postischemic hearts, however, led to dramatic improvement in myocardial function in both groups (p < 0.05 for all parameters) and significant improvement in oxygen consumption (p = 0.01). CONCLUSIONS: In a normal, isolated, blood-perfused pig heart subjected to 30 minutes of total normothermic ischemia, (1) enrichment of the perfusate with aspartate/glutamate before and after ischemia affects neither myocardial energy metabolism during ischemia-reperfusion nor postischemic recovery of myocardial function or oxygen consumption and (2) inotropic stimulation can recruit significant postischemic function and sufficient aerobic respiration to support it, irrespective of aspartate/glutamate enrichment. PMID- 9202689 TI - Prevention of the hypoxic reoxygenation injury with the use of a leukocyte depleting filter. AB - OBJECTIVES: Recent studies have shown that an injury occurs when the hypoxic heart is suddenly reoxygenated (as occurs with cardiopulmonary bypass), resulting in myocardial depression, impaired oxygenation, and increased pulmonary vascular resistance. We hypothesize that this injury is, in part, due to oxygen-derived radicals produced by activated white cells and may therefore be ameliorated by limiting leukocytes in the bypass circuit. METHODS: Fifteen neonatal piglets underwent 60 minutes of ventilator hypoxia (inspired oxygen fraction 8% to 10%), followed by reoxygenation with cardiopulmonary bypass at an inspired oxygen fraction of 100% for 90 minutes. In nine piglets (group 1) our routine bypass circuit was used with no modifications, and in six piglets (group 2) a leukocyte depleting filter (Pall BC-1; Pall Biomedical Products Corporation, Glencoe, N.Y.) was inserted in the arterial line to lower the neutrophil count. Six additional piglets underwent 90 minutes of bypass without hypoxia (cardiopulmonary bypass controls). Postbypass myocardial and pulmonary function was assessed by pressure volume loops, arterial/alveolar ratio, and pulmonary vascular resistance index. Results are expressed as a percentage of control. RESULTS: By comparison with group 1 piglets (reoxygenation without a filter), hypoxic piglets undergoing reoxygenation with a leukocyte-depleting filter (group 2) had improved myocardial systolic function (88% vs 52%; p < 0.05), diastolic compliance (175% vs 275%; p < 0.05), and preload recruitable stroke work (91% vs 54%; p < 0.05); had better preservation of the arterial/alveolar ratio (97% vs 74%; p < 0.05); and had less increase in pulmonary vascular resistance (229% vs 391%; p < 0.05). Furthermore, leukocyte filtration prevented adenosine triphosphate depletion or a change in tissue antioxidants. Conversely, unprotected piglets (group 1) exhibited lower levels of adenosine triphosphate and significant loss of tissue antioxidants. Indeed, the results in the leukocyte-filtered piglets (group 2) were nearly identical to those of piglets subjected to bypass without hypoxia (controls). CONCLUSIONS: (1) This study demonstrates that a major component of the injury that occurs when the hypoxic heart is abruptly reoxygenated is caused by oxygen radicals produced by white blood cells; (2) this injury can be prevented by a leukocyte-depleting filter; and (3) avoidance of this injury improves postbypass myocardial and pulmonary function. These data suggest that leukocyte depletion should be used routinely in all children undergoing operations for cyanotic heart disease or extracorporeal membrane oxygenation. PMID- 9202690 TI - Systolic ventricular interaction in normal and diseased explanted human hearts. AB - OBJECTIVE: The purpose of this study was to quantify the magnitude of interaction between the right and left ventricles in conditions of heart failure. METHODS: Human hearts were taken from transplant recipients diagnosed with diluted cardiomyopathy at the time of transplantation and were restored to beating condition with use of an isolated perfusion circuit. Left ventricular-right ventricular interaction was determined by ramping volume in the left ventricle while holding right ventricular volume constant. Right ventricular pressure gain was plotted against left ventricular pressure and the slope of the linear regression determined the left ventricular-right ventricular interaction. A similar procedure was used to determine right ventricular-left ventricular interaction. Two normal hearts were obtained from transplant donors not suitable for cardiac donation to serve as control hearts. RESULTS: Mean left ventricular right ventricular interaction was 0.22 in the hearts with dilated cardiomyopathy compared with 0.06 in the control hearts. Mean right ventricular-left ventricular interaction was 0.14 in the hearts with dilated cardiomyopathy compared with 0.09 in the control hearts. A marked increase in left ventricular-right ventricular interaction was noted in the hearts with dilated cardiomyopathy compared with control hearts. Although observed values of right ventricular-left ventricular interaction also correspond to previously published results, no significant increase was observed in the dilated cardiomyopathy condition. CONCLUSIONS: These studies confirm previously published values for systolic ventricular interaction obtained with animal models and demonstrate a marked increase in the dependence of the right ventricle on left ventricular function to maintain systolic pressure generation during conditions of dilated cardiomyopathy. PMID- 9202691 TI - Deoxygenated blood minimizes adherence of sonicated albumin microbubbles during cardioplegic arrest and after blood reperfusion: experimental and clinical observations with myocardial contrast echocardiography. AB - Both administration of cardioplegic solution and blood reperfusion result in endothelial dysfunction. The transit rate of albumin microbubbles during myocardial contrast echocardiography may reflect endothelial injury. Accordingly, we performed myocardial contrast echocardiography in 12 dogs undergoing cardiopulmonary bypass and measured the myocardial transit rate of microbubbles injected into the aortic root during delivery of cardioplegic solutions containing arterial and venous blood and delivery of pure crystalloid cardioplegic solution. The myocardial transit rate of 99mTc-labeled red blood cells was measured and perfusates were sampled for biochemical analysis at each stage. The microbubble transit rate was markedly prolonged during delivery of crystalloid cardioplegic solution and improved significantly during infusion of blood cardioplegic solution (p < 0.001); venous compared with arterial blood in the solution resulted in a greater rate (p < 0.001). The microbubble transit rate did not correlate with pH, oxygen tension or carbon dioxide tension values, or K+ concentration. The red blood cell transit rate remained constant regardless of the cardioplegic perfusate infused. Myocardial contrast echocardiography was also performed in 12 patients undergoing coronary artery bypass who underwent sequential arterial and venous reperfusion after cardioplegic arrest. The microbubble transit rate was faster with venous than arterial blood reperfusion (p = 0.01), although this gain was diminished when arterial blood reperfusion preceded venous blood reperfusion (p = 0.05). Our results indicate that endothelial dysfunction after cardioplegic arrest may be ameliorated by reperfusion with venous rather than arterial blood. PMID- 9202692 TI - Early failure of freehand aortic stentless xenograft valves. PMID- 9202693 TI - Immunolocalization of basic fibroblast growth factor receptors in internal thoracic artery and saphenous vein grafts. PMID- 9202694 TI - Transapical aortic cannulation for hypothermic aortic operation through a left thoracotomy: an alternative to avoid retrograde arterial perfusion. PMID- 9202695 TI - Heparin-associated intrathoracic pseudotumor. PMID- 9202696 TI - Single leukocyte filter (Pall BC1B) fails in multidose cold blood cardioplegia. PMID- 9202697 TI - Improvement of outcomes after coronary artery bypass. PMID- 9202698 TI - Subxiphoid tube drainage in bullectomy and lung volume reduction: a word of caution. PMID- 9202699 TI - Assessment of the hemodynamic performance of small-size aortic valve prostheses. PMID- 9202700 TI - Myocardial protection of the hypoxic heart. PMID- 9202702 TI - Twenty-three-year survival of a bovine pericardial bioprosthesis. PMID- 9202701 TI - Use of epsilon-aminocaproic acid to reduce bleeding. PMID- 9202703 TI - Nathan Green (1871-1955). PMID- 9202704 TI - Capture-ELISA: a new assay for the detection of immunoglobulin M isotype antibodies using Chlamydia trachomatis antigen. AB - We present here a new method of IgM antibody-capture enzyme-linked immunosorbent assay (IgM-Capture-ELISA) for the diagnosis of recently acquired infections with Chlamydia trachomatis. For this analysis, plates were coated with goat IgG anti human Fc mu. The capture of serum IgM antibodies was revealed indirectly by the sequential addition of biotinylated chlamydial proteins and peroxidase-conjugated streptavidin. In chlamydial extracts, cysteine-rich proteins are preferential antigenic targets for the humoral response. 3-(N-maleimidopropionyl)-biocytin (MPB), which binds biotinylated moieties to sulfhydryl groups, was used for the labeling procedure. The preservation of the antigenic specificity of labeled proteins was controlled by a blotting of these proteins, which were, respectively, probed either with specific IgM antibodies or with streptavidin. This analysis revealed that, after labeling, recognized epitopes are more particularly present on the major outer membrane protein (MOMP) of Chlamydia trachomatis. The validation of IgM-Capture-ELISA was assessed by using 170 selected sera from patients suspected of being infected by Chlamydia. Results were respectively compared to conventional indirect immunofluorescence assays (MIF-IgM assays) and to Western blotting. Sixteen sera were found to possess IgM antibodies against Chlamydia trachomatis with IgM-Capture-ELISA. Among these 16 sera, 14 and 15 were, respectively, positive with MIF-IgM assays and in Western blotting. Data obtained with IgM-Capture-ELISA reveal the absence of false positive results in sera containing rheumatoid factor, which has been shown to interfere in the two other methods. IgM-Capture-ELISA value was then confirmed using sera from patients consulting for genital or pulmonary diseases, from patients with confirmed chlamydial infections, and from patients with other pathologies. IgM-Capture-ELISA appears as an alternative simple semi-quantitative assay for the detection of early chlamydial infection. PMID- 9202705 TI - Purification of immunogenic heat shock protein 70-peptide complexes by ADP affinity chromatography. AB - Adenosine triphosphate (ATP)-affinity chromatography has been widely used to purify molecules of the Hsp70 family. This procedure leads to dissociation of peptides from Hsp70 molecules, resulting in Hsp70 preparations devoid of immunological activity. We report that substitution of ATP-affinity chromatography by ADP-affinity chromatography results in isolation of Hsp70 molecules which are still associated with peptides and are immunogenic. The Hsp70 preparations thus obtained contain the constitutive Hsp73 and the inducible Hsp72 molecules. These observations furnish a basis for an analysis of the structural heterogeneity among the members of the Hsp70 family and of the antigenic peptides associated with individual members of this family. They also provide a practical method for the isolation of large quantities of immunologically active Hsp70 peptide preparations. PMID- 9202706 TI - Generation of antibodies to human IL-12 and amphiregulin by immunization of Balb/c mice with diepitope multiple antigen peptides. AB - Six peptide sequences derived from the human proteins/oligopeptides IL-12, amphiregulin and FALL-39 were synthesized in order to raise specific antibodies in Balb/c mice. Although peptides are valuable tools for generating specific antibodies, they are often poor immunogens due to their small size and lack of relevant T-cell epitopes. To circumvent these limitations, the human peptides were co-synthesized in diepitope multiple antigen peptides (MAP) with a known H 2d-restricted T helper-cell epitope. The importance of including a T-cell epitope in the diepitope MAPs was demonstrated by the fact that only one of the human peptides was immunogenic as a monoepitope MAP, lacking the T-cell epitope. Conversely, all diepitope MAPs generated potent antibody responses to the desired human peptides as well as to the T-cell epitope. A certain degree of variability of the antibody responses to the diepitope MAPs indicated that the alterable component, i.e. the human B-cell epitope, influenced the T-cell help elicited by the T-cell epitope. Still, the relative conformity of the B-cell responses suggests that this strategy is generally applicable for a rational production of specific antibodies. Moreover, antiserum to four diepitope MAPs recognized the corresponding full-length human protein/oligopeptide as did monoclonal antibodies made against IL-12-and amphiregulin-based MAPs. PMID- 9202707 TI - Selection of phage-displayed superantigen by binding to cell-surface MHC class II. AB - We have expressed the superantigen staphylococcal enterotoxin A (SEA) on the surface of bacteriophage as a fusion with the gene VIII protein (gVIIIp). This phage-displayed superantigen retains the properties inherent in the natural protein. It binds to MHC class II and activates T-cells bearing appropriate V beta regions. A flexible 5-amino acid linker sequence between the SEA molecule and the phage coat protein improved the production of functional phage-displayed SEA. Binding to MHC class II-expressing cells effectively selected SEA-phage from non-SEA-phage background. This indicates that this will be an effective method for selecting new specificities of superantigen from libraries of SEA mutants and for cloning of novel superantigens. PMID- 9202708 TI - Purification and binding properties of a human ficolin-like protein. AB - Ficolin was initially identified from porcine uterus as a TGF-beta 1 binding protein and is considered to have an overall structure similar to that of the complement protein C1q and the collectins. Recent studies have shown that human ficolin is synthesized mainly by monocytes in peripheral blood and that it could potentially bind to sugar structures on microorganisms. The aim of the present investigations was to isolate ficolin from human plasma by affinity chromatography on immobilized sugars. A human serum protein was identified in the GlcNAc eluate from GlcNAc-Sepharose which migrated as a polypeptide of approx. 40 kDa on SDS-PAGE under reducing conditions and was, after further purification by FPLC on a mono-Q column, shown to have an identical N-terminal sequence, over the first 14 residues, to P35, a plasma protein having similar sequence and domain organisation to ficolin. This protein, named the ficolin-like protein, was shown to be sensitive to collagenase and similar to P35 in that it was also disulphide linked into an oligomer of approx. 320 kDa. However, unlike P35, its binding to GlcNAc was independent of Ca2+. Gel-filtration studies showed that this ficolin like protein also had a molecular weight of approx. 320 kDa under non dissociating conditions. During the course of this study this ficolin-like protein was found to simply bind to CNBr-activated Sepharose which had been inactivated with Tris, and from which it could be eluted with GlcNAc. This ficolin-like protein was also shown to bind to GlcNAc, but not to mannose and maltose. The functional significance of the unusual binding property of this ficolin-like protein is not clear, but it has facilitated the development of a simple method for its purification. PMID- 9202709 TI - A flow cytometric method for the rapid detection of beta-galactosidase transfected cells: an in vitro and in vivo study. AB - A flow cytometric method has been developed for the rapid analysis of lacZ transduced cells. The method described is based on an indirect immunofluorescence staining procedure using a monoclonal antibody which binds specifically to beta galactosidase from E. coli and to beta-galactosidase fusion proteins. This technique was used for the quantification in vitro as well as in vivo of beta galactosidase expression in B16 melanoma cells. The described method is appropriate for a variety of cell types (species, lineage), is simple, quantitative, reliable, rapid and applicable to all constructs containing the lacZ selectable markers. It should prove to be very helpful (1) for the quantification of cells expressing the lacZ reporter gene and (2) for studying gene regulation, including transfection modality, promoter efficacy, enhancer activity, and other regulatory factors. PMID- 9202710 TI - Comparison of cytokine measurements using ELISA, ELISPOT and semi-quantitative RT PCR. AB - We have investigated the correlation between results obtained by three different methods (semi-quantitative RT-PCR, ELISA and ELISPOT) used to measure cytokine expression by mouse leukocytes. The production of the cytokines tumour necrosis factor-alpha (TNF alpha), interferon-gamma (IFN gamma) and interleukin-4 (IL-4), was analysed with all three methods. In a simple experimental murine in vivo model of leukocyte stimulation, consisting of a single intravenous injection of anti-CD3 antibodies followed by a short incubation in vitro, the results obtained with spleen cells for each of the three cytokines differed greatly, depending on the method used. For TNF alpha, a significant increase in RNA was observed upon stimulation, whereas the number of spot-forming cells did not increase and the protein was not detectable in serum or in cell culture supernatants by ELISA. In vitro cultured splenocytes showed a strong correlation between all three methods for IFN gamma. Upon stimulation, the amount of RNA for IL-4 increased in parallel with the secretion of the cytokine and the number of spot-forming cells. However, high numbers of spot forming cells were observed in controls. We conclude that, depending on the specific aim of an investigation, combinations of different methods have to be chosen carefully in order to detect activation of leukocytes for cytokine expression. PMID- 9202711 TI - Purification and characterization of secretory IgA from baboon colostrum. AB - In this report, we describe a method for purifying secretory immunoglobulin A (sIgA) from baboon (Papio anubis) colostrum. The colostrum was first clarified by centrifugation and then analyzed with various anti-human Ig-specific immunologic reagents. Cross-reactive IgA in the baboon colostrum was identified by ELISA. Western blot analysis also demonstrated cross-reactive epitopes associated with human IgA1, IgA2, secretory component (SC), and joining (J) chain. To purify the sIgA, colostrum was separated into 4 distinct fractions by gel filtration chromatography. Analysis of the individual fractions by ELISA indicated that the IgA elutes over one peak. The IgA fraction was compared with purified human sIgA on SDS-PAGE, and exhibited heavy (H) chains, light (L) chains, SC, and J chain. The baboon colostrum was also analyzed by ELISA for specific IgG H and L chain epitopes utilizing monoclonal antibodies (MAbs). No significant quantity of IgG was detected in the baboon colostrum or in the individual 4 fractions, while L chain reactivity was observed in the sIgA fraction. The sIgA fraction was pooled, concentrated, and was found to contain approximately 7 mg/ml sIgA. To determine if the baboon sIgA was dimeric like human sIgA, the purified sIgA was sized by molecular sieve chromatography. The molecular size of the sIgA preparation (350 kDa) was determined empirically by comparison to known molecular species used to calibrate the column. In addition, native SDS-PAGE indicated that baboon sIgA, like human sIgA, migrates between IgG and IgM, suggesting it has a dimeric form. The purified baboon sIgA preparation should prove useful in the future study of mucosal immune responses induced in non-human primate species and for the generation of sIgA-specific immunological reagents. PMID- 9202712 TI - Versatile vectors for transient and stable expression of recombinant antibody molecules in mammalian cells. AB - We have developed new cassette expression vectors for the cloning of any intact V region gene followed by any C-region gene. Both the heavy-and light chain vectors harbor a strong hCMV promoter, restriction site cassettes for cloning of both V- and C-region genes, transcription termination signals, fl-ori for single stranded DNA (ssDNA) synthesis, selection marker for Neomycin and SV40 ori for transient expression. The vectors accept VH and VL chain genes obtained by RT-PCR. Reamplification of the V genes is then performed with a new set of primers which are designed specifically for each individual V gene. Cloning into the vectors is aided by restriction sites located just outside the V-gene coding region, thus keeping the V-genes intact. The vectors also contain cloning sites for the exchange of genomic C-genes so that the resulting Ig genes may code for complete antibodies, antibody fragments or fusion proteins. A simple subcloning step permits the expression of both heavy and light chain genes from one single vector, thus avoiding co-transfection of the two vectors. The usefulness of the vectors was confirmed by construction of mouse-human chimeric antibodies. The V genes were derived from a hybridoma cell line, TP-3, and was combined with human C kappa, C gamma 3 and C gamma 1 genes as well as with CH1 gamma 3. High yields of recombinant antibody products in NSO cells were obtained. Transient expression was also demonstrated. PMID- 9202713 TI - A highly sensitive cytotoxicity assay based on the release of reporter enzymes, from stably transfected cell lines. AB - The well-established methods of generating stably transfected cell lines, and the detection of nanomolar amounts of an enzyme in a fast and reproducible assay, were utilised to establish non-radiometric cytotoxicity assays. In these assay systems, the detection of released enzymes was used to quantitate the leakage of intracellular proteins after membrane disintegration. Target cell lines were transfected with a luciferase reporter gene under the control of a strong eucaryotic promoter. Release of the intracellular expressed enzyme into the culture supernatant occurred after membrane perforation and was measured as an indicator of cellular death. The quantitation of released enzyme was a reliable indicator of cell death initiated either by complement-mediated killing, or by cell-mediated cytotoxicity. This system was initially established with P815 mastocytoma cells as an example of a target cell line. Transfection with the firefly luciferase gene provided an intracellular enzyme absent in mammalian cells. In a parallel approach, P815 and BW5147 target cells were transfected with bacterial beta-galactosidase to provide a similar cytotoxicity system. This enzyme, however, has a considerably longer half life in tissue culture medium than luciferase. In a direct comparison between the standard 51Cr release and beta-galactosidase release, the enzyme release showed a much higher signal-to noise ratio, i.e., low background and high induced release if spontaneous release and detergent induced maximal lysis were measured. Since a wide range of human and murine cell lines can be stably transfected and several reporter genes are available, the system should provide an alternative for conventional cytotoxicity assays. The detection of released enzymes by colorimetric or luminometric methods makes this cytotoxicity assay independent of radionuclides. The sensitivity of luminometric enzyme detection systems should also permit the measurement of apoptotic processes and might make in vivo studies of cellular death using transgenic animals feasible. PMID- 9202714 TI - CHO transfectants produce large amounts of recombinant protein in suspension culture. AB - Chinese hamster ovary (CHO) cells transfected with various genes are widely used as adherent cell monolayers to produce recombinant proteins. In this report we present a new culture technique for CHO cells transfected with the vector pPOL DHFR-CD14 using a minifermenter (miniPERM, Heraeus) for the production of recombinant human endotoxin receptor CD14 (rCD14). The transfectants were cultured for 12-17 days under serum-free conditions and formed spheroids. From this system we harvested supernatants containing up to 3.1 mg/ml recombinant CD14 (rCD14). This represents a 200-fold increase of rCD14 yield compared to conventional adherent CHO cell culture. PMID- 9202715 TI - Perspectives on Canadian field studies examining the potential of pulp and paper mill effluent to affect fish reproduction. AB - The results and interpretations of published Canadian field studies on the reproductive status of fish in waters receiving pulp and paper mill effluent discharges were reviewed. Most of the information was obtained from indicator measurements such as gonad size, fecundity, and serum steroid levels in wild fish sampled at reference and effluent-exposed sites. Difficulties in selecting appropriate sampling sites, natural variability, and the ecological relevance of the indicator measurements were identified as major complicating factors for the interpretation of the field data. Consequently, it was not possible to conclude to what extent, if any, widespread effects on fish reproduction are being caused by pulp and paper mill effluents or that specific manufacturing processes are causing such effects. Further research on the normal variability and predictive capability of reproductive indicators, for example, using an integrated approach (i.e., laboratory testing, mesocosm studies, and field work), is recommended. PMID- 9202716 TI - Ozone-induced DNA strand breaks In guinea pig tracheobronchial epithelial cells. AB - Ozone (O3), the major oxidant of photochemical smog, is thought to be genotoxic and a potential respiratory carcinogen or promoter of carcinogenic processes. Because of oxidative reactions with the mucus in the upper airway, O3 reaction products are able to penetrate into the tracheobronchial epithelial (TE) cells. The carcinogenic effects of O3 on the TE cells are especially of interest since most previous studies have focused on the morphology or permeability changes of tracheas only. Therefore, the objective of this study was to examine the potential O3 genotoxicity in TE cells after an in vivo exposure, using DNA strand breaks as an index. Two-month-old male Dunkin-Hartley guinea pigs, specific pathogen free, 4 in each group, were exposed to 1.0 ppm O3 for 0, 12, 24, 48, 72, or 96 h. Animals exposed to filtered air without O3 exposure were used as controls. After O3 exposure, the trachea with two main bronchi was removed from each animal, and TE cells were isolated and employed for determination of DNA strand breaks by fluorometric analysis of DNA unwinding (FADU). The statistical significance level was set at alpha = .05. Compared with controls, ozone exposure did not alter the TE cell yield or viability, but caused an increase in protein content in tracheal lavage and an increase in DNA strand breaks. The amount of DNA left in the alkali lysate of TE cells found at 72 h exposure was significantly decreased from controls for 3 different alkali incubation times. An increase of the double-stranded DNA left in the alkali lysate of TE cells was observed at 96 h of exposure and approached the value of 24 h of exposure. The same pattern was seen with all 3 different alkali incubation times at 15 degrees C. One Qd unit was estimated to correspond to 100 strand breaks per cell. The Qd was also used as an indicator for O3 damage. Compared to controls, the Qd increases significantly after 1 ppm O3 exposure for 72 h, regardless of the alkali incubation time at 15 degrees C. PMID- 9202717 TI - Growth-related signaling in vascular smooth muscle cells is deregulated by TCDD during the G0/G1 transition. AB - Experiments have been conducted to examine the impact of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) on growth-related signaling in vascular smooth muscle cells (SMCs). A 40% reduction of peak DNA synthesis was observed in SMCs only when TCDD was added during the G0/G1 transition of the cell cycle. Enhanced phosphorylation of several endogenous proteins during this period was coincident with increased tyrosine kinase activity as early as 15 min following TCDD challenge. No changes in protein phosphorylation status occurred in cells treated with TCDD during the G1/S transition or during S phase. Cotreatment of quiescent SMCs with 10 nM TCDD and serum for 3 h reduced serum-inducible binding activity to a 12-O-tetradecanoyl phorbol 13-acetate responsive element (TRE) by approximately 40%. No alterations of constitutive TRE binding were observed in quiescent SMCs treated with TCDD for up to 5 h. These data show that mitogen related signaling in vascular SMCs is modulated by TCDD selectively during the G0/G1 transition, and these effects influence the growth behavior of these cells. PMID- 9202718 TI - Systemic toxicity of dermally applied crude oils in rats. AB - Two crude oils, differing in viscosity (V) and nitrogen (N) and sulfur (S) content, were evaluated for systemic toxicity. In the Crude I (low V, low N, low S) study, the material was applied to the clipped backs of rats at dose levels of 0, 30, 125, and 500 mg/kg. In the Crude II (high V, high N, moderate S) study, the oil was applied similarly at the same dose levels. The crude oils were applied for 13 wk, 5 d/wk. Exposure sites were not occluded. Mean body weight gain (wk 1-14) was significantly reduced in male rats exposed to Crude II; body weight gain of all other animals was not adversely affected by treatment. An increase in absolute (A) and relative (R) liver weights and a decrease in A and R thymus weights were observed in male and female rats exposed to Crude II at 500 mg/kg; only liver weights (A and R) were adversely affected in male and female rats exposed to Crude I. In general, there was no consistent pattern of toxicity for serum chemistry endpoints; however, more parameters were adversely affected in Crude II-exposed female rats than in the other exposed groups. A consistent pattern of toxicity for hematology endpoints was observed among male rats exposed to Crude I and male and female rats exposed to Crude II. Parameters affected included: Crudes I and II, red blood cell count, hemoglobin, and hematocrit; Crude II, platelet count. Microscopic evaluation of tissues revealed the following treatment-related findings: Crude I, treated skin, thymus, and thyroid; Crude II, bone marrow, treated skin, thymus, and thyroid. The LOEL (lowest observable effect level) for skin irritation and systemic toxicity (based on marginal effects on the thyroid) for both crude oils was 30 mg/kg; effects were more numerous and more pronounced in animals exposed to Crude II. Systemic effects are probably related to concentrations of polycyclic aromatic compounds (PAC) found in crude oil. PMID- 9202719 TI - Biliary elimination of oral 2,4-dichlorophenoxyacetic acid and its metabolites in male and female Sprague-Dawley rats, B6C3F1 mice, and Syrian hamsters. AB - The role of biliary elimination in the metabolic disposition of 2,4-D was evaluated in male and female Sprague-Dawley rats, B6C3F1 mice, and Syrian hamsters. Following cannulation of the bile duct, an intragastric (ig) dose of 2,4-D (200 mg/kg) was administered and bile was collected at 30- or 60-min intervals for up to 6 h. Bile flow rates were constant in rats, increased in mice, and decreased in hamsters throughout the collection periods. Total recovery of radioactivity was greatest in male mice (about 7% of administered dose over 4 h). Female mice and rats of both sexes excreted about 3% over the same interval and male and female hamsters about 1%. About 71-88% of the activity in bile was parent compound. The glycine conjugate of 2,4-D was found in bile from mice, rats, and hamsters and the taurine conjugate in bile from mice. The only sex dependent difference in the metabolite profile was in mice. Male mice excreted twice as much glycine conjugate as female mice. An additional minor metabolite (4 7%) was present in rat and mouse bile. This was tentatively identified as 2,4-D glucuronide based on its hydrolysis by beta-glucuronidase. One more very minor metabolite (3%) was detected in rat bile but was not characterized due to its lability. The results of this study indicate that there are species-dependent differences in the biliary elimination of 2,4-D but not sex-dependent differences. PMID- 9202720 TI - Two potential clinical applications of the alkaline single-cell gel electrophoresis assay: (1). Human bladder washings and transitional cell carcinoma of the bladder; and (2). Human sperm and male infertility. AB - Part 1: The alkaline single-cell gel electrophoresis (comet) assay was used to analyse the integrity and DNA content of exfoliated cells extracted from bladder washing specimens from 9 transitional cell carcinoma patients and 15 control patients. DNA damage, as expressed by % tail DNA and tail moment values, was observed to occur in cells from both control and bladder cancer samples. The extent of the damage was, however, found to be significantly greater in the cancer group than in the control group. Comet optical density values were also recorded for each cell analysed in the comet assay and although differences observed between tumour grades were not found to be statistically significant, the mean comet optical density value was observed to be greater in the cancer group than in the control population studied. These preliminary results suggest that the comet assay may have potential as a diagnostic tool and as a prognostic indicator in transitional cell carcinoma. Part 2: Baseline DNA damage in sperm cells from 13 normozoospermic fertile males, 17 normozoospermic infertile males and 11 asthenozoospermic infertile males were compared using a modified alkaline comet assay technique. No significant difference in the level of baseline DNA damage was observed between the 3 categories of sperm studied; however the untreated sperm cells were observed to display approximately 20% tail DNA. This is notably higher than the background DNA damage observed in somatic cells where the % tail DNA is normally less than 5%. Sperm from the 3 groups of men studied were also compared for sensitivity to DNA breakage, using the modified alkaline comet assay, following X-ray irradiations (5, 10 and 30 Gy) and hydrogen peroxide treatments (40, 100 and 200 microM). Significant levels of X-ray-induced damage were found relative to untreated control sperm in the two infertile groups following 30 Gy irradiation. Significant damage in hydrogen peroxide-treated sperm was observed in sperm from fertile samples, at 200 microM and in infertile samples at 100- and 200-microM doses relative to controls. These results therefore indicate that fertile sperm samples are more resistant to X-ray- and hydrogen peroxide-induced DNA breakage than infertile samples. Further studies involving greater numbers of individuals are currently in progress to confirm these findings. PMID- 9202721 TI - Induction of DNA damage by risk factors of colon cancer in human colon cells derived from biopsies. AB - In order to increase the understanding of the factors responsible for causing human colon cancer, a technique was developed to detect genotoxic effects of chemicals in human colon cells. Risk factors suspected to be associated with the aetiology of human colon cancer were subsequently investigated: the method is based on the measurement of DNA damage in primary cells freshly isolated from human colon biopsies with the single cell microgel ectrophoresis technique ('Comet Assay'). 2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 2-amino 3-methyl-3H-imidazo[4,5f]quinoline (IQ), N-methyl-N-nitro-N-nitrosoguanidine (MNNG), dinitrosocaffeidine (DNC) lithocholic acid (LCA), hydrogen peroxide (H2O2) and benzo[a]pyrene (B[a]P) were investigated for their genotoxic and cytotoxic effects following 30 min incubation with colon cells of human, and for comparative purposes also of the rat colon. The nitrosamides (MNNG, DNC) were very genotoxic in human colon cells. MNNG was more genotoxic in human than in rat colon cells. In contrast, the rat colon carcinogens PhIP and IQ were not genotoxic in human colon cells. PhIP did induce DNA damage in rat colon cells, which correlates to its capacity of inducing tumors in this animal tissue. LCA was toxic (rat > human) and concomitantly caused DNA damage in higher concentrations. The widespread contaminant B[a]P was not genotoxic in colon cells of either species using this system. H2O2 was found to be a potent genotoxic agent to both rat and human colon cells (human > rat). In summary, those compounds chosen as representatives of endogenously formed risk factors (MNNG, H2O2, LCA) have a higher toxic and/or genotoxic potency in human colon tissue than in rat colon. They are also more effective in this system than the contaminants tested so far (B[a]P, PhIP, IQ). The newly developed technique is rapid and yields relevant results. It is a novel and useful approach to assess different chemical compounds for genotoxic activities in tumour target tissues of the human. PMID- 9202722 TI - Evaluation of DNA damage in leukocytes of ex-smokers by single cell gel electrophoresis. AB - Single cell gel electrophoresis (SCGE), or comet assay, appears to be a promising tool to estimate DNA damage at the single cell level and it provides information on the presence of damage among individual cells. A follow-up study of 90 smokers who ceased smoking was undertaken to determine the possible decrease of DNA damage in their leukocytes. Before beginning the trial, volunteers smoked on average 26.1 +/- 8.4 cigarettes/day. Comet length did not correlate with the number of cigarettes/day or with the condensate tar content. At the end of the study, 28 volunteers had abandoned the trial, 40 volunteers relapsed into smoking at different times, but with a reduced number of cigarettes/day, whereas 22 fully succeeded in smoking cessation. Throughout the 5 sampling times, a great variability of comet length at individual level was found. However, after 1 year of follow-up, comet length means were found to be significantly shorter (p < 0.0001) in those volunteers who completely quit smoking compared to those who relapsed into smoking (27.2 +/- 1.6 vs. 31.9 +/- 5.1 microns, respectively), irrespective of the amount of cigarettes previously smoked. No effect of age or sex was found. Six months later, these results were confirmed by a further study carried out on a reduced sample of volunteers. The present data strongly suggest that, in spite of the great variability observed, 1 year of smoking cessation is associated with a significant reduction of DNA damage in circulating leukocytes. PMID- 9202723 TI - The genetic toxicity of time: importance of DNA-unwinding time to the outcome of single-cell gel electrophoresis assays. AB - Single-cell gel electrophoresis assays (comet assays) are described in which DNA damage is assessed in mouse skin keratinocytes treated with N-methyl-N'-nitro-N nitrosoguanidine (MNNG) and beta-propiolactone (BPL) either in vitro or in vivo. The positive results observed under both conditions of test encourage the further development of the mouse skin comet assay as a screen for direct-acting in vivo genotoxins. From the outset of the present experiments we were struck by the compacted nature of the DNA in mouse skin keratinocytes. Under similar conditions of assay, rodent hepatocytes presented a uniform 'unwound' distribution of DNA over the whole nuclear region. In order to study this effect we varied what seemed to be the most obviously related assay parameter: the DNA-unwinding time. A series of experiments was conducted in which control and MNNG-treated cells were exposed to a range of alkaline DNA-unwinding times (0.3-18 h) followed by measurement of the three comet tail parameters (length, DNA content, and their product, tail moment). Each of these parameters increased with increasing time of unwinding such that the tails observed for MNNG-treated cells with 0.3 h of DNA unwinding were similar in length to the tails of control cells exposed to an 8 h DNA-unwinding time. It is concluded that DNA-unwinding time is a critical parameter of the comet assay and that it may require optimisation for each tissue/cell type studied. Further, the data alert to the prospect that agents that uniquely affect chromosomal protein superstructure may increase comet tail length/DNA content in the absence of chemically induced DNA damage. Thus, there may be two discrete classes of chemical interaction with chromosomal DNA that yield identical comet assay results, but which have different implications for the genetic toxicity of the test agent. Similar effects were observed for rat hepatocytes or mouse lymphoma cells exposed to an 18 h DNA-unwinding time, but no comet tails were produced by exposure of cells to the lysis conditions (pH 10.0) for 18 h. PMID- 9202724 TI - Use of the comet assay to investigate possible interactions of nitric oxide and reactive oxygen species in the induction of DNA damage and inhibition of function in an insulin-secreting cell line. AB - We have previously used the comet assay to demonstrate that the nitric oxide donor 3-morpholinosydnonimine (SIN-1) produces DNA damage in rat islets of Langerhans and in the SV40-transformed insulin-secreting hamster cell line, HIT T15. Damage is not prevented by the addition of superoxide dismutase (SOD). In the present study, we have compared SIN-1, which generates nitric oxide, superoxide anion and hydrogen peroxide, with two other nitric oxide donors, S nitrosoglutathione (GSNO) and the tetra-iron-sulphur cluster nitrosyl, Roussin's black salt (RBS). We have used the comet assay as a highly sensitive method to measure DNA-damaging ability, and also measured inhibition of DNA synthesis and inhibition of insulin secretion. We have examined the effect of SOD and catalase on each of these endpoints in HIT-T15 cells following a 30-min exposure to the compounds (24 h for DNA synthesis). All compounds produced a significant dose dependent increase in strand-breakage formation and all inhibited DNA synthesis and glucose-stimulated insulin secretion. RBS was the most potent. SOD did not reduce the responses observed with any of the compounds. Catalase largely prevented DNA strand breakage, inhibition of DNA synthesis and inhibition of insulin secretion by SIN-1, but had no effect on responses to GSNO or RBS. Addition of SOD together with catalase gave no greater protection against SIN-1 than catalase alone. The nitric oxide and superoxide anion produced by SIN-1 are though to combine to form highly reactive peroxynitrite. In addition, H2O2 may be formed in the presence of SIN-1 and may form hydroxyl radical in the presence of a transition metal, such as Fe2+. It appears that in insulin-secreting cells, the effects of SIN-1 are largely mediated by this latter mechanism. In contrast, GSNO and RBS appear to act by a different mechanism, not overtly involving reactive oxygen species. GSNO and H2O2 show no significant interaction in the induction of DNA strand breaks. Both nitric oxide and H2O2 are effective, directly or indirectly, as DNA strand-breaking agents, inhibitors of DNA synthesis and inhibitors of insulin secretion. PMID- 9202725 TI - Evaluation of the genotoxic activity of metronidazole and dimetridazole in human lymphocytes by the comet assay. AB - The genotoxicity of metronidazole (MZ) and dimetridazole (DZ) has been evaluated in human lymphocytes using the comet assay. The test has been performed using 3 doses (58.4, 175.2 and 292.1 microM for MZ; and 70.9, 212.6 and 354.3 microM for DZ) under 3 experimental protocols: aerobiosis, anaerobiosis (90% N2, 10% CO2) and with the presence of the microsomal fraction S9 mix. The effects of 4 antioxidants (8-hydroxyquinoline (8HQ), vitamin C (VitC), catalase (CAT) and superoxide dismutase (SOD), have been investigated on DNA damage generated by fixed concentrations of MZ (292.1 microM) and DZ (354.4 microM). In aerobic conditions, MZ and DZ produced significant dose-response relationships. The dose related effects of both drugs decreased or were abolished in anaerobic conditions or in presence of S9 mix. 8HQ, VitC, CAT and SOD induced dose-related protective responses against DNA damage due to MZ and DZ. These findings suggest that MZ and DZ induce DNA damage in human lymphocytes through the futile cycle. The one electron reduction of the drugs leads to the production of nitro radical anions. In the presence of oxygen, these radicals are reoxidized and generate oxygen activated species. PMID- 9202726 TI - Detection of subpopulations resistant to DNA-damaging agents in spheroids and murine tumours. AB - Chinese hamster V79 monolayers, V79 spheroids, and SCCVII murine tumours were examined for DNA damage using the alkaline comet assay and for cell killing by measuring clonogenicity following a 1-h exposure to doxorubicin, N-methyl-N' nitro-N-nitrosoguanidine (MNNG), 4-nitroquinoline-N-oxide (4-NQO), etoposide, or 3-amino-1,2,4-benzotriazine-1,4-dioxide (tirapazamine). Greater heterogeneity in DNA damage was evident in spheroids compared to monolayers exposed to these drugs, and cell survival was correlated with the fraction of cells which lacked sufficient DNA damage following treatment with tirapazamine or doxorubicin. Cell sorting experiments verified that subpopulations of cells resistant to DNA damage were also more resistant to cell killing. Significant heterogeneity was observed in cells from SCCVII tumours exposed to tirapazamine and etoposide, and comet DNA content was used to independently assess DNA damage to aneuploid tumour cells and diploid host cells. These results suggest that, for some drugs, the comet assay may be an effective method of identifying drug-resistant cells in solid tumours. PMID- 9202727 TI - The DNA 'comet assay' as a rapid screening technique to control irradiated food. AB - The exposure of food to ionizing radiation is being progressively used in many countries to inactivate food pathogens, to eradicate pests, and to extend shelf life, thereby contributing to a safer and more plentiful food supply. To ensure free consumer choice, irradiated food will be labelled as such, and to enforce labelling, analytical methods to detect the irradiation treatment in the food product itself are desirable. In particular, there is a need for simple and rapid screening methods for the control of irradiated food. The DNA comet assay offers great potential as a rapid tool to detect whether a wide variety of foodstuffs have been radiation processed. In order to simplify the test, the agarose single layer set-up has been chosen, using a neutral protocol. Interlaboratory blind trials have been successfully carried out with a number of food products, both of animal and plant origin. This paper presents an overview of the hitherto obtained results and in addition the results of an intercomparison test with seeds, dried fruits and spices are described. In this intercomparison, an identification rate of 95% was achieved. Thus, using this novel technique, an effective screening of radiation-induced DNA fragmentation is obtained. Since other food treatments also may cause DNA fragmentation, samples with fragmented DNA suspected to have been irradiated should be analyzed by other validated methods for irradiated food, if such treatments which damage DNA cannot be excluded. PMID- 9202728 TI - The comet assay: what can it really tell us? AB - A range of applications of the alkaline comet assay is covered, from investigations of the physicochemical behaviour of DNA, through studies of cellular responses to DNA damage, to biomonitoring of human populations. The underlying principles of this assay are discussed, and new evidence presented which supports the concept of relaxation of supercoiled loops, rather than alkaline unwinding, as the primary reason for comet tail formation. DNA-damaging agents that do not induce strand breaks directly can be detected when cellular repair processes convert lesions to transient strand breaks; an approach is outlined here which maximises this effect and thus widens the scope of the assay. Purified repair enzymes, applied to DNA during the course of the comet assay procedure, greatly increase the sensitivity and specificity of the assay; recent developments with formamidopyrimidine glycosylase (recognising 8-OH-gua and other damaged purines) and uvrABC (for bulky lesions) are presented. The kinetics of cellular repair after low doses of oxidative damage have been followed with this modified comet assay. Finally, the successful measurement of biomarkers of oxidative damage in human populations establishes the comet assay as a valuable tool in molecular epidemiology. PMID- 9202729 TI - Sodium ascorbate induces DNA single-strand breaks in human cells in vitro. AB - Incubation of human lymphocytes, neonatal fibroblasts, and Molt-4 cells (T-cell leukemia cell line) with sodium ascorbate for 1 h resulted in a concentration dependent increase in DNA single-strand breaks as assayed by an alkaline microgel electrophoresis technique. Fibroblasts and Molt-4 cells were significantly more sensitive than lymphocytes to the induction of DNA single-strand breaks by 25, 50, and 100 microM concentrations of sodium ascorbate. Significant cell loss was observed in Molt-4, but not in lymphocyte and fibroblast cultures, after 4 h of incubation in 50 microM of sodium ascorbate, a concentration similar to the plasma level of ascorbic acid in humans. PMID- 9202730 TI - Assessment of DNA damage induced in vitro by etoposide and two fungicides (carbendazim and chlorothalonil) in human lymphocytes with the comet assay. AB - The effects of two fungicides (carbendazim and chlorothalonil) on the induction of DNA damage in human peripheral blood lymphocytes (human PBL) have been investigated using the single cell gel electrophoresis assay (SCGE assay or comet assay) immediately after a 1-h treatment and after a 24-h post-treatment incubation. The assessment of etoposide (an effective antitumour agent) effects on human PBL in terms of cell viability and dose-DNA damage relationships was made and etoposide selected as a positive control. The results indicate that etoposide induces significant (p < 0.01) dose-dependent DNA damages for concentrations at which the loss of cell viability is low. After a 24-h recuperation period, all observed DNA damages has disappeared. With SCGE assay performed after a 1-h treatment, similar positive results were observed with chlorothalonil alone or in association with carbendazim, without any loss of cell viability. However, a dramatic loss of cell viability was measured after 24 h and was associated with a large proportion of highly damaged cells. In contrast, carbendazim was not cytotoxic on human PBL and did not induced DNA damage using the SCGE assay either immediately after treatment or after a 24-h post-treatment incubation. These results point to the necessity of an adequate evaluation of immediate and long-term cytotoxicity of compounds that are to be assessed by the SCGE assay. PMID- 9202731 TI - Sixth International Conference on Carcinogenic/Mutagenic N-Substituted Aryl Compounds. Monterey, California, November 4-8, 1995. Proceedings. PMID- 9202732 TI - Binding of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) to protein- and low molecular weight thiols and its role in ring hydroxylation. AB - The N-oxidized species of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) have been shown to react with thiols. We have previously characterized a glutathione conjugate of PhIP linked via the C2 of PhIP with apparent loss of the amino group, in rat hepatocytes and PhIP exposed rats. This metabolite was possibly formed from 1-methyl-2-nitro-6-phenylimidazo[4,5-b]pyridine (nitro PhIP). Upon reacting nitro-PhIP with rat albumin, both in the presence and absence of a reducing system, four products were observed after enzymic proteolysis. One of them was markedly increased after 2-mercaptoethanol treatment of the protein. This adduct was linked to a cysteine-S via C2 of PhIP. Using N2 acetoxy-PhIP as a starting material, an unstable protein adduct was observed which degraded to 50% of the original concentration (t 1/2) after 3 days. This is compatible with the finding that serum PhIP adducts decline rapidly in PhIP exposed rats. Unstable adducts were also formed following the reaction of N2 acetoxy-PhIP with glutathione or cysteine. Based on mass spectroscopy and UV spectra analysis, the suggested structures were RS(-S-)-(H)N2-PhIP. In all cases a degradation product identified as 5-hydroxy-PhIP was formed as characterized by mass spectrometry and NMR spectroscopy. 5-hydroxy-PhIP and its glucuronyl derivative were also observed in rat hepatocytes incubated in vitro with PhIP. In bile of PhIP-exposed rats, only the glucuronyl derivative was observed. Depletion of glutathione reduced the amount excreted in bile and experiments with microsomes indicate that hydroxylation directly at the 5 position is not mediated by cytochrome P-450 mono-oxygenase system. This indicates that 5-hydroxy-PhIP may be formed from N-acetoxy-PhIP via binding to thiols also in cells. PMID- 9202733 TI - Arylamine-DNA adduct conformation in relation to mutagenesis. AB - A considerable body of evidence has indicated that local conformational alterations induced by DNA adducts may provide the molecular basis for differences in mutational specificities exhibited by structurally similar adducts. To elucidate the relationships between adduct structure and mutation induction, the ability of several single-ring arylamines present in tobacco smoke (i.e., methylanilines, dimethylanilines, and ethylanilines) to form DNA adducts was investigated. In all cases, the major adducts were C8-substituted deoxyguanosine derivatives, which is consistent with what has been observed with more carcinogenic arylamines, such as 2-aminofluorene and 4-aminobiphenyl. Spectroscopic and theoretical data on the adducts indicated conformational differences depending upon the location of the alkyl substituents. Adducts containing alkyl groups ortho to the amino function (e.g., 2-methylaniline) had a greater percentage of syn conformers about the glycosyl bond than those not bearing such groups. Arylamines with ortho alkyl substituents tend to be more mutagenic and tumorigenic than analogues not containing an ortho alkyl substituent. This increase in biological activity may be due in part to the greater propensity of ortho alkylated adducts to adopt a syn conformation. PMID- 9202734 TI - Comparative metabolism and DNA binding of 1-, 2-, and 4-nitropyrene in rats. AB - The metabolism and DNA binding studies of mono-NP isomers under identical conditions were conducted, as an initial investigation, in order to provide an understanding for the higher carcinogenic activity of 4-NP in the rat mammary gland. Urinary and fecal excretion patterns of 4-NP and 1-NP 24 h following administration to female CD rats (i.p.; 24 mg/kg body weight; 1.55 mCi/rat) were similar but higher than those of 2-NP. The identified metabolites were formed via nitroreduction and ring oxidation pathways. Neither the excretion patterns nor the nature of the metabolites readily explained why the mammary tumorigenic activity of these three isomers varied. Although overall levels of mono-NP bound to liver DNA did not account for the observed differences in the biological activity, further HPLC analysis of the liver DNA hydrolysates showed that only 4 NP had yielded putative multiple DNA adducts; none were detected in the case of 1 NP and 2-NP. 1-, 2-, and 4-NP were found to bind to mammary DNA at levels of 0.6, 0.3, and 2.1 pmol/mg DNA, respectively. The structure of DNA adducts in the mammary gland and in the liver of female CD rats following the i.p. administration of 4-NP has not been identified. Collectively, the results of this preliminary study indicate that the difference in levels of DNA binding in the mammary gland in vivo may reflect why 4-NP has higher carcinogenic activity. PMID- 9202735 TI - Determination of heterocyclic aromatic amines in food products: automation of the sample preparation method prior to HPLC and HPLC-MS quantification. AB - Heat-processing protein-rich foods may cause the formation of heterocyclic aromatic amines (HAAs), all of which have mutagenic and some also carcinogenic potential. Accurately measuring HAA levels in food products is therefore a necessary to realistically assess this risk factor. A solid-phase extraction method for quantitative HAA analysis has been developed by us over the last few years. This method has recently been automated using a robotic workstation and now allows almost unattended sample preparation, a process which saves a human operator about five hours of benchwork. Cleaned-up samples were analyzed by high performance liquid chromatography (HPLC) and ultraviolet (UV) or mass spectrometric (MS) detection. While HPLC-UV remains the daily tool to quantify HAAs, we found HPLC-electrospray-MS to be an alternative detection method with unique advantages, suited for both HAA identification and quantification. PMID- 9202737 TI - Metabolism of isomeric nitrobenzo[a]pyrenes leading to DNA adducts and mutagenesis. AB - We have been interested in determining the structural and electronic features that may be useful in predicting the mutagenic activity of nitro-polycyclic aromatic hydrocarbons (nitro-PAHs). We have previously found that a correlation between structural and electronic features and direct-acting mutagenicity in Salmonella typhimurium cannot be made using nitro-PAHs with different molecular size. In this study, a series of structurally related nitro-PAHs, the environmental contaminants 1-, 3-, and 6-nitrobenzo[alpha]pyrene (NBaP) and their derivatives, was used to determine structure-activity relationships. It was found that isomeric NBaPs are activated to DNA damaging and mutagenic derivatives by nitroreduction, ring-oxidation, or by a combination of these two pathways. A general finding was that NBaPs and derivatives with their nitro substituent oriented perpendicular to the aromatic system exhibit either very weak or no direct-acting mutagenicity in S. typhimurium strains TA98 and TA100. In this paper, we also discuss the effect of the location of the nitro group on the metabolism and the mutagenicity of NBaPs and the effect of oxygen-containing functional groups on the mutagenicity of NBaP derivatives. These findings provide a useful molecular basis for interpreting and predicting the direct-acting mutagenicity of nitro-PAHs. PMID- 9202736 TI - Health risks of heterocyclic amines. AB - Common cooking procedures such as broiling, frying, barbecuing (flame-grilling), heat processing and pyrolysis of protein-rich foods induce the formation of potent mutagenic and carcinogenic heterocyclic amines. These same compounds produce tumors at multiple organ sites in both mice and rats. One example of these induced tumors has also been seen in nonhuman primates. Risk assessment for the human population consuming these compounds requires the integration of knowledge of dosimetry, metabolism, carcinogenic potency, and epidemiology. When this integration is done in even a preliminary way as is done here, the range of risk for an individual from these compounds is enormous. Exposure contributes a range of 200-fold or more and metabolism and DNA repair differences among individuals could easily be an additional 10-fold between individuals. This indicates that differences in human cancer risk for heterocyclic amines could range more than a thousandfold between individuals based on exposure and genetic susceptibility. PMID- 9202738 TI - Assessing human risk to heterocyclic amines. AB - Heterocyclic amines such as MeIQx and PhIP are potent genotoxic chemicals which are formed at part per billion levels when meat is cooked. Using assays based on gas chromatography/mass spectrometry with stable isotope labelled analogues as internal standards we have demonstrated MeIQx and PhIP, are efficiently absorbed into the systemic circulation after ingestion of fried beef. Using a potent and selective inhibitor of human CYP1A2, furafylline, we have shown that N hydroxylation catalysed by this enzyme is the major pathway of metabolism of MeIQx and PhIP and is solely responsible for their oxidation to mutagenic species. This is in contrast to the situation in laboratory animals in which both activation by N-hydroxylation and deactivation by C-oxidation occurs. When furafylline was administered to human volunteers before ingestion of fried beef, we showed that > 90% of MeIQx and approximately 70% of PhIP elimination could be inhibited, demonstrating the extent to which activation occurred in man. MeIQx is a very powerful mutagen in bacterial assays whereas PhIP is a more potent mammalian cell mutagen. Using a mammalian cell target gene, hprt, we have shown that PhIP induces a characteristic mutational 'fingerprint' which is identical to that detected in the Apc gene of 5/8 colonic tumours induced by PhIP in rats. These studies support a biological association between HA exposure and diet related tumours but emphasise that information obtained from animal studies does not always reflect the situation in humans. PMID- 9202739 TI - Human acetyltransferase polymorphisms. AB - Conjugation of primary amino and hydroxylamino groups with acetate, catalyzed by acetyl CoA-dependent arylamine acetyltransferase (NAT) enzymes, may play an important role in the intricate series of metabolic pathways that produce or prevent toxicity following exposure to homo- and heterocyclic arylamine and hydrazine xenobiotics. Two independently regulated and kinetically distinct human acetyltransferases are now known to exist, namely NAT1 and NAT2. Interindividual variation in NAT2 function is associated with the classical isoniazid acetylation polymorphism which was discovered over forty years ago. At last count, fifteen variant alleles at the NAT2 gene locus have been linked to the isoniazid 'acetylator phenotype', and each of these can be identified in population studies using specific PCR-based genotyping tests. On the other hand, NAT1 shows kinetic selectivity for compounds whose disposition is unrelated to the classical isoniazid acetylation polymorphism. NAT1 expression is also phenotypically variable in human populations, at least in part due to allelic differences at the NAT1 gene locus. Nine NAT1 variant alleles have been described to date, of which NAT1* 14 and NAT1* 15 clearly produce defective NAT1 proteins and lead to functional impairment in the metabolism of NAT1-selective substrates both in vivo and in vitro. On the other hand, it has been reported that the NAT1* 10 variant associates with elevated NAT1 activity and increased risk for cancers of the bladder and colon. Because of the important toxicologic consequences of allelic variation in NAT1 and NAT2 function for the metabolic activation of arylamine and heterocyclic amine procarcinogens, further studies are needed to improve our understanding of the extent of NAT allelic variation, to determine the functional capacity of each variant gene product, and to develop accurate methods of detecting them in population and epidemiological studies. PMID- 9202740 TI - Molecular mechanisms of mutagenesis by aromatic amines and amides. AB - The mutagenic properties of 2-acetylaminofluorene-derived DNA adducts, including N-(deoxyguanosin-8-yl)-2-acetylaminofluorene, N-(deoxyguanosin-8-yl)-2 aminofluorene, N-(deoxyguanosin-N2-yl)-2-acetylaminofluorene, and several minor oxidation products have been explored, using site-specific techniques. Oligodeoxynucleotides containing a single AAF-derived DNA adduct were prepared by postsynthetic modification and used as templates in primer extension reactions catalyzed by bacterial and mammalian DNA polymerases. Base substitutions and deletions occurring during DNA synthesis were quantified. dG-C8-AAF promoted one- and two-base deletions and small amounts of incorporation of dCMP, dAMP, and/or dTMP opposite the lesion in reactions catalyzed by the 3'-->5' exonuclease-free Klenow fragment of DNA polymerase 1 (exo-) and polymerase alpha. dG-C8-AF did not miscode in reactions catalyzed by exo-; however, base misincorporation and deletions were observed in reactions with pol alpha. dG-N2-AAF promoted small amounts of dAMP incorporation in reactions catalyzed by exo-. The miscoding potential of minor oxidation products of dG-C8-AF was much higher than that of other adducts. Steady-state kinetics were used to measure frequencies of nucleotide insertion opposite the lesion and chain extension from the 3' terminus. Kinetic data were consistent with the results of primer extension studies. A mutation 'hot spot' was constructed and the influence of sequence context on the frequency of deletions generated by dG-C8-AAF was explored systematically in reactions catalyzed by exo-. Based on our results with aminofluorene DNA adducts, we propose a general mechanism for frameshift deletion mutagenesis. Site-specific methods also were used to establish the mutagenic potential of AAF-derived DNA adducts in mammalian cells. dG-C8-AAF and dG-C8-AF exhibited similar mutagenic specificities, predicting the occurrence of G-->T transversions and G-->A transitions in mammalian cells. PMID- 9202741 TI - Cytochrome P450 in the breast and brain: role in tissue-specific activation of xenobiotics. AB - It is still an open question as to whether, upon administration of procarcinogens to rodents, development of cancers in extrahepatic tissues is due to activation of these chemicals in the liver or to in situ activation within the tissue. The low level of P450 in many tissues means that it is very difficult to demonstrate the formation of significant amounts of reactive metabolites when these tissues are incubated with procarcinogens in vitro. It is our contention that the importance of tissue-specific activation of procarcinogens can best be decided when the cells which harbour P450 have been identified and the isozyme profile in the cells defined. With this aim in view, we have begun to characterize the forms of P450 in the breast and brain. Perhaps not surprisingly, the P450s in the breast are regulated as a function of age and hormonal status of rats and most of the breast P450 can be accounted for by hepatic forms. The P450 content of the brain, on the other hand, is very responsive to environmental factors. The quantity of P450 as well as the isozyme profile is altered by drugs and chemicals in the environment. The P450s induced in the brain are similar to liver P450s, but the constitutive forms are not. P450s of the 1A family are inducible in both tissues and this indicates that heterocyclic amines can be activated in the brain and polycyclic aromatic hydrocarbons in the breast. The cells in which this activation can occur remain to be identified. PMID- 9202742 TI - Quantitative structure-activity (QSAR) relationships of mutagenic aromatic and heterocyclic amines. AB - We extended our previous studies of mutagenic/carcinogenic heterocyclic aromatic amines formed during the cooking of foods to 66 aromatic and 99 heterocyclic amines for which mutagenic potency data are available. The amines require activation by enzymes to form metabolites reactive with DNA and exhibit an enormous range of potency as frameshift mutagens in the Ames/Salmonella assay. To ascertain factors that might influence potency, structural features and quantum mechanical parameters calculated by the Huckel method (and, for a subset of 20 amines, by semi-empirical AM1, and ab initio methods) were analyzed by multiple linear regression. The major findings were: (1) earlier findings on cooked food mutagens and their synthetic congeners can be extended to other amines; (2) mutagenic potency is directly related to the number of fused aromatic rings (size of the aromatic system), the number of ring nitrogen atoms (participation of lone electron pairs in the pi-cloud), and presence of a methyl substituent on a ring nitrogen; (3) potency is inversely related to the energy of the lowest unoccupied molecular orbital (LUMO) of the parent amine. Ford and Griffin (1992) and Sabbioni and Wild (1992) showed that the LUMO energy of the derived nitrenium ion is closely related to its stability (calculated with reference to aniline). Increased stability has been hypothesized to enhance the probability of adduct formation with DNA by avoiding detoxifying side reactions and increasing the lifetime of the ion. In the large heterogeneous series of amines in our present study the Huckel method energy of the highest occupied molecular orbital (HOMO), rather than the LUMO energy, of the nitrenium ion was marginally related to the potency of the parent amine. However, in the selected subset of 20 amines with ab initio calculation, the LUMO energy of the ion confirmed the previous reports. The contribution of quantum chemical factors to mechanistic insight on the mutagenicity and carcinogenicity of aromatic and heterocyclic amines is still under development. PMID- 9202743 TI - Antigenotoxic activity of natural chlorophylls. AB - Chlorophyllin, a man-made water-soluble form of chlorophyll, is a focus of intensive studies from many laboratories for its antimutagenic and anticarcinogenic properties. Natural chlorophylls, in contrast, have been little studied in this regard. Since yellow-green vegetables are implicated to be protective against human cancers by epidemiological studies, it is important to explore the antigenotoxic properties of natural chlorophylls. Previously, we reported that a chlorophyll sample prepared from Chlorella vulgaris inhibited the mutagenicity of 3-hydroxyamino-1-methyl-5H-pyrido[4,3-b]indole, a direct-acting mutagen, in Salmonella, and that the chlorophyll also showed inhibition of wing spot formation in Drosophila induced by 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). We have now prepared several samples of chlorophyll from spinach and chlorella, and studied their effect on the genotoxicity of 4-nitroquinoline 1 oxide (4NQO) in Drosophila. The results showed that the genotoxicity of orally given 4NQO was suppressed by simultaneous administration of the chlorophylls. The mechanisms of this inhibition are discussed. PMID- 9202744 TI - Rodent models of the human acetylation polymorphism: comparisons of recombinant acetyltransferases. AB - The acetylation polymorphism is associated with differential susceptibility to drug toxicity and cancers related to aromatic and heterocyclic amine exposures. N Acetylation is catalyzed by two cytosolic N-acetyltransferases (NAT1 and NAT2) which detoxify many carcinogenic aromatic amines. NAT1 and NAT2 also activate (via O-acetylation) the N-hydroxy metabolites of aromatic and heterocyclic amine carcinogens to electrophilic intermediates which form DNA adducts and initiate cancer. The classical N-acetylation polymorphism is regulated at the NAT2 locus, which segregates individuals into rapid, intermediate, and slow acetylator phenotypes. Some human epidemiological studies associate slow acetylator and rapid acetylator phenotypes with increased susceptibility to urinary bladder and colorectal cancers, respectively. The acetylation polymorphism has been characterized in three rodent species (mouse, Syrian hamster, and rat) to test associations between NAT2 acetylator phenotype and susceptibility to aromatic and heterocyclic amine-induced cancers in various tumor target organs. NAT1 and NAT2 from rapid and slow acetylator mouse, Syrian hamster, and rat each have been cloned and sequenced. Recombinant NAT1 and NAT2 enzymes enzymes encoded by these genes have been characterized with respect to their catalytic activities for both activation (O-acetylation) and deactivation (N-acetylation) of aromatic and heterocyclic amine carcinogens. The acetylation polymorphisms in mouse, Syrian hamster, and rat are herein reviewed and compared as models of the human acetylation polymorphism. PMID- 9202746 TI - Chemoprotection against the formation of colon DNA adducts from the food-borne carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in the rat. AB - The mutagenic heterocyclic aromatic amine, 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP), is a pyrolysis product in cooked foods that has been shown to be a rat colon carcinogen and has been implicated in the etiology of human colon cancer. In order to identify chemoprotection strategies that could be carried out in humans, a pilot study was conducted in which PhIP-DNA-adduct levels were quantified in the colons of male F344 rats that had been subjected to 16 different putative chemoprotection regimens, followed by a gavage of PhIP (50 mg/kg) and sacrifice 24 h later. The 16 treatments (Oltipraz, benzylisothiocyanate, diallyl sulfide, garlic powder, ethoxyquin, butylated hydroxyanisole, glutathione, indole-3-carbinol, alpha-angelicalactone, kahweol/cafestol palmitates, quercetin, green tea, black tea, tannic acid, amylase-resistant starch, and physical exercise) comprised sulfur-containing compounds, antioxidants, flavonoids, diterpenes, polyphenols, high dietary fiber, etc. The strongest inhibition of PhIP-DNA adduct formation in the colon was observed upon pretreatment with black tea, benzylisothiocyanate, and a mixture (1:1) of kahweol:cafestol palmitates, which resulted in 67, 66, and 54% decreases in colon PhIP-DNA adduct levels, as compared with controls. Preliminary studies on their mechanism of action indicated that only kahweol:cafestol caused a substantial induction of glutathione S-transferase isozymes (GSTs) that are thought to be important in the detoxification of PhIP. Notably, this induction occurred in the liver rather than in the colon. PMID- 9202745 TI - Carcinogenicity of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) in the rat. AB - A total of 10 highly-mutagenic heterocyclic amines have been identified to be carcinogenic in rodents. Among these, 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP), generally the most abundant with normal cooking procedures, induces mammary and colon carcinomas in rats in a clear dose-dependent manner. In a two-generation exposure (transplacental and trans-breast milk) experiment using Sprague-Dawley rats, an increased risk of mammary adenocarcinoma development was found in the second generation. Excretion of PhIP into the milk and transfer of PhIP to fetuses and neonates with resultant hepatic PhIP-DNA adduct formation were also confirmed. On the other hand, PhIP mammary carcinogenesis was significantly inhibited by coadministration of chlorophyllin or a synthetic antioxidant, 1-O-hexyl-2,3,5-trimethylhydroquinone, in long-term experiments using female F344 rats. The available findings strongly suggest that this food derived carcinogen might be of importance as an environmental factor in the production of human cancers and that its carcinogenicity could be largely avoided by reducing intake of such compounds or by adoption of appropriate chemopreventive measures. PMID- 9202747 TI - Role of acetyltransferases in the metabolism and carcinogenicity of aromatic amines. AB - The genotoxicity of N-substituted aryl compounds is dependent on their conversion to reactive metabolites, frequently through the production of reactive N acetoxyarylamines. This activation is accomplished by acetyltransferases that are widely distributed. In the rat, the production of N-acetoxyarylamines has been most clearly related to the induction of tumors in the mammary gland, but this pathway also appears to be an important factor in the production of tumors in the liver, Zymbal gland and gastrointestinal tract. Expression of rat acetyltransferases responsible for acetylation of the nitrogen and the oxygen of arylamine derivatives (i.e., acetyltransferases 1 and 2) in bacterial cells has now permitted experiments which demonstrate that these enzymes exhibit good affinities for and N-acetylation of the endogenous arylalkylamines derived from tryptophan, i.e., tryptamine, 5-hydroxytryptamine (serotonin) and 5 methoxytryptamine, the immediate metabolic precursor of melatonin. Evidence that these reactions are likely to reflect real biological potentials is bolstered by histological localization of acetyltransferase mRNAs with synthetic antisense oligodeoxynucleotide probes. The results of these studies in rat indicate that the expression of acetyltransferase in tissues of the central nervous, gastrointestinal, urinary and reproductive systems is highly regulated, as it is in other organs commonly associated with aromatic amine carcinogenicity. Similar experimental approaches have been successful with human liver, mammary gland, kidney and bladder preparations. These observations give evidence that genotoxic N-acetoxyarylamines are produced by acetyltransferases that can metabolize, and possibly modulate, the hormonal and neurotransmitter effects of endogenous arylalkylamines. These relationships may help explain the occasional induction of tumors in organs not usually considered as targets of aromatic amines, as well as raise the possibility that the production of N-oxidized endogenous substrates may represent a mechanism for tumor induction in the absence of exogenous carcinogens. PMID- 9202748 TI - Analysis of cooked muscle meats for heterocyclic aromatic amine carcinogens. AB - A number of related heterocyclic amines that are mutagenic in bacterial test systems and carcinogenic in animals are formed during the cooking of food. The most commonly reported and abundant compounds are PhIP, MeIQx, DiMeIQx, IQ and A alpha C. Using analysis by solid-phase extraction and HPLC, amounts found in foods range from less than one ng/g for products from fast-food restaurants, up to 14 ng/g in commercially cooked products and over 300 ng/g for well done flame grilled chicken breast meat. Interestingly, marinating meat for 4 h greatly reduces the amount of PhIP produced during cooking, but not MeIQx. Comparing mutagenic activity in meat samples to the mutagenic activity accounted for by the known heterocyclic amines shows that most samples have activity that cannot be accounted for by the aromatic amines we can currently identify. This suggests that additional compounds are present in these foods and need to be investigated, particularly those grilled over open flames. PMID- 9202749 TI - Polymorphisms of CYP1A1 and GSTM1 influence the in vivo function of CYP1A2. AB - Differences in human cancer susceptibility have been attributed to polymorphisms of carcinogen metabolizing enzymes. Our efforts have focused on the systems responsible for metabolism of aromatic and heterocyclic amines found in cigarette smoke and in cooked foods. Cytochrome P4501A2 (CYP1A2), which catalyzes aromatic and heterocyclic amine N-oxidation, has been implicated as a risk factor in both urinary bladder and colorectal cancer. In the present study we used the results of caffeine phenotyping experiments to measure the effects of cigarette smoke and compounds present in meat cooked at high temperature on CYP1A2 activity. Subjects in the smoking cessation study had mean CYP1A2 activity of 17.8 (expressed as the urinary molar ratio of [17X + 17U]/137X) while smoking: however, this activity decreased to 10.9 three weeks after cessation of smoking. Subjects in the cooked meat feeding study had mean CYP1A2 activity of 9.01 after 1 week of consuming meat cooked at low temperature, but this value increased to 12.7 after 1 week of consuming meat cooked at high temperature. Because no association has been identified between differences in CYP1A2 activity and variations in the CYP1A2 structural gene, we sought to determine whether the activities of other carcinogen metabolizing enzymes are involved in the regulation of CYP1A2 activity. CYP1A2 activity was higher in individuals who express the GSTM1 null allele compared to those expressing the GSTM1*A,B allele, 10.2 vs. 8.5 for unexposed conditions and 15.0 vs. 12.3 for exposed conditions. CYP1A1 genotyping demonstrated that individuals possessing the Ile/Ile CYP1A1 genotype had greater mean CYP1A2 activity than those who had the heterozygous Ile/Val allelic variant of the CYP1A1 gene. However, upon exposure to cigarette smoke or high-temperature cooked meat, individuals possessing the heterozygous form of the CYP1A1 gene had significantly increased CYP1A2 activity (18.1) compared to those with the more common Ile/Ile CYP1A1 genotype (13.3). These results indicate that CYP1A2, CYP1A1, and GSTM1 gene-gene interactions could be important confounders in the interpretation of molecular epidemiology studies. PMID- 9202750 TI - Relationship between adduct formation, rates of excision repair and the cytotoxic and mutagenic effects of structurally-related polycyclic aromatic carcinogens. AB - The cytotoxic and mutagenic effect of 1-nitrosopyrene (1-NOP) and N-acetoxy-2 acetylaminofluorene (N-AcO-AAF) were compared with that of (+/-)-7 beta, 8 alpha dihydroxy-9 alpha, 10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BPDE) as a function of the initial frequency of adducts formed in the DNA of repair proficient diploid human fibroblasts and the fraction remaining at the time the cells replicate their DNA. The principal adducts of all three agents involve guanine. The initial level of BPDE-, 1-NOP-, or N-AcO-AAF-induced adducts per 10(6) nucleotides required to lower the survival of these cells to 37% of the control was 8, 25, and 50, respectively. The frequency of mutants per 10(6) clonable cells induced at those levels of initial adduct formation was 160, 80, and 40, respectively. We determined the rate of excision repair of these adducts from the overall genome, from the individual strands of the hypoxanthine phosphoribosyltransferase (HPRT) gene, and in the case of 1-NOP and BPDE, at the level of individual nucleotides in the nontranscribed strand of exon 3 of that gene, a region where mutations induced by those agents are particularly frequent. 1-NOP-induced adducts were excised from the overall genome and from the individual strands of HPRT at a rate 2-3 times faster than BPDE-induced adducts. The average rate of repair of 1-NOP-induced adducts in exon 3 was also 2-3 times faster than the average rate of repair of BPDE-induced adducts. However, at particular nucleotides 1-NOP-induced adducts were repaired much faster, or slower, or in some cases at a rate equal to that of BPDE-induced adducts. Excision repair of N-AcO-AAF-induced adducts (i.e., deacetylated aminofluorene residues) was significantly slower than that of BPDE- or 1-NOP-induced adducts, and was not strand-specific. In an in vitro assay, BPDE adducts were four times more effective in blocking transcription than were 1-NOP or N-AcO-AAF-induced adducts. We conclude that the cytotoxic and mutagenic effect of these carcinogens reflect a complex interplay of adduct conformation, ability of adducts to block replication and transcription, and variation in the rate of excision repair, even at the nucleotide level. PMID- 9202751 TI - The role of xenobiotic metabolizing enzymes in arylamine toxicity and carcinogenesis: functional and localization studies. AB - In both animal models and humans, the first and obligatory step in the activation of arylamines is N-hydroxylation. This pathway is primarily mediated by the phase I enzymes CYP1A1, CYP1A2 and CYP4B1. In the presence of flavonoids such as alpha naphthoflavone and flavone, both CYP3A4 and CYP3A5 have also been shown to play a minor role in the activation of food-derived heterocyclic amines. The further activation of N-hydroxyarylamines by phase-II metabolism can involve both N, O acetylation and N, O-sulfonation catalyzed by N-acetyltransferases (NAT1 and NAT2) and sulfotransferases, respectively. Using an array of techniques, we have been unable to detect constitutive CYP1A expression in any segments of the human gastrointestinal tract. This is in contrast to the rabbit where CYP1A1 protein was readily detectable on immunoblots in microsomes prepared from the small intestine. In humans, CYP3A3/3A4 expression was detectable in the esophagus and all segments of the small intestine. Northern blot analysis of eleven human colons showed considerable heterogeneity in CYP3A mRNA between individuals, with the presence of two mRNA species in some subjects. Employing the technique of hybridization histochemistry (also known as in situ hybridization), CYP4B1 expression was observed in some human colons but not in the liver or the small intestine. Hybridization histochemistry studies have also demonstrated variable NAT1 and NAT2 expression in the human gastrointestinal tract. NAT1 and NAT2 mRNA expression was detected in the human liver, small intestine, colon, esophagus, bladder, ureter, stomach and lung. Using a general aryl sulfotransferase riboprobe (HAST1), we have demonstrated marked sulfotransferase expression in the human colon, small intestine, lung, stomach and liver. These studies demonstrate that considerable variability exists in the expression of enzymes involved in the activation of aromatic amines in human tissues. The significance of these results in relation to a role for heterocyclic amines in colon cancer is discussed. PMID- 9202752 TI - Genetic changes induced by heterocyclic amines. AB - Clarification of the mutational fingerprints of HCAs offers a promising approach in the investigation of the role of heterocyclic amines (HCAs) in human carcinogenesis. We analyzed mutations in the tumor related genes of tumors induced by HCAs, 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), 2-amino-3 methylimidazo[4,5-f]quinoline (IQ), and 2-amino-1-methyl-6-phenylimidazo[4,5 b]pyridine (PhIP), which mainly yield DNA-adducts of C8-guanine. The G-->T transversion at codon 13-2nd position in Ha-ras was predominantly observed in mouse forestomach and rat Zymbal gland tumors induced by MeIQ. In contrast, various types of mutation were detected in the ras family genes of rat Zymbal gland tumors induced by IQ; the presence of a methyl group at position 4 of imidazo[4,5-f]quinoline gave rise to a remarkable difference in the mutational fingerprint. Apc mutations were detected in PhIP- and IQ-induced rat colon tumors, with incidences of 50% (4/8) and 15% (2/13), respectively. All five mutations detected in the four PhIP-induced tumors consisted of a guanine deletion from the 5'-GGGA-3' sequence, in contrast with T to C and C to T mutations in IQ-induced tumors. Four of these five mutations shared seven common nucleotides, -GTGGGAT- surrounding the guanine; indicating that PhIP leaves a characteristic mutational fingerprint in Apc. Colon tumors induced by PhIP were also found to have mutations in their microsatellite sequences, and similar results were detected in mammary gland tumors induced by PhIP, contrasting with no mutations in IQ-induced colon tumors and a very low frequency of mutations in 7,12-dimethylbenz[a]anthracene (DMBA)-induced mammary tumors. Although the mechanisms involved in the induction of microsatellite mutations are not known yet, microsatellite mutations which can also be detected in sporadic human tumors, including colon and breast tumors, were indicated to be a characteristic of PhIP. Mammary tumors induced by PhIP showed loss of heterozygocity (LOH) at the distal part of chromosome 10, which shows synteny with the distal part of human chromosome 17, where LOH frequently occurs in human breast cancer. In conclusion, each heterocyclic amine leave a mutational fingerprint which is specific to each compound. Since the tumor-related genes involved in PhIP-induced tumors have characteristics in common with those in human cancers, further detailed analysis will provide us with useful information on mutational fingerprints, and on the possible contribution of PhIP to human colon cancer. PMID- 9202753 TI - The dual role of 2-acetylaminofluorene in hepatocarcinogenesis: specific targets for initiation and promotion. AB - 2-Acetylaminofluorene (AAF) is one of the most widely studied model carcinogens. It produces liver tumors in rats. Comparison with other arylamides shows that promutagenic DNA lesions are necessary but not sufficient to explain this tissue specific effect. Mutagenicity of AAF was studied in AS52 cells and compared with that of 2-acetylaminophenanthrene and trans-4-acetylaminostilbene which are incomplete carcinogens in rat liver. The major mutations were G to T transversions in all cases. All three acetamides acted as initiators in an initiation-promotion experiment with phenobarbital as a promoter. Chronic toxic effects of AAF were attributed to specific effects of AAF metabolites on mitochondrial respiration. Electron drainage by 2-nitrosofluorene causes an uncoupling effect on oxidative phosphorylation in vitro. Corresponding compensatory effects were observed in vivo. Initiating as well as promoting properties of AAF are therefore considered responsible for the generation of rat liver tumors. The results support the hypothesis that genotoxic effects generate initiated cells which begin to proliferate only when microcirculation is disturbed due to cirrhotic alterations. These are triggered by non-genotoxic interference with mitochondrial respiration and oxidative phosphorylation. PMID- 9202754 TI - Aromatic amine DNA adduct formation in chronically-exposed mice: considerations for human comparison. AB - Lifetime chronic exposure of mice to the aromatic amines 4-aminobiphenyl (ABP) and 2-acetylaminofluorene (AAF) produces liver and urinary bladder tumors. In parallel experiments, DNA adduct levels in target tissues reach a steady-state (a balance between adduct formation and removal) after about four weeks of either AAF or ABP ingestion. For these and other carcinogens, steady-state DNA adduct levels most frequently increase linearly with dose, but the formation of tumors also depends upon a variety of factors, including the proliferative capacity of the target tissue, the sex of the animal, genotoxic properties of the specific adducts formed, and other unknown events. Chronic dosing experiments in animal models are of interest for human risk assessment because human exposure is typically intermittent, involving repeated exposures. However, it is to be expected that in a genetically-diverse human population, where the lifetime averages > 70 years, the relationship between tumorigenesis and DNA adduct formation will be relatively more complex than that observed in mice. From our studies of chronic ABP exposure in male mice, we have obtained the daily dose of ABP and the steady-state level of N-(deoxyguanosin-8-yl)-4-aminobiphenyl (dG-C8 ABP) adduct associated with a 50% mouse bladder tumor incidence. Our attempt at a human extrapolation for adducts and urinary bladder cancer in smoking males (20 40 cigarettes/day) is based on the ABP dose per cigarette, values for the dG-C8 ABP adduct in bladder biopsies of lifetime heavy smokers at age approximately 70, and the smoking-related bladder tumor incidence (absolute lifetime risk) for Caucasian males in the United States aged 65-84 years. The extrapolation has produced two major predictions, one related to adduct formation and the other related to tumorigenesis. First, the observed level of smoking-related dG-C8-ABP in DNA of human bladder epithelium, expressed as a function of daily ABP intake, is about 3500-times higher than similar data for mice, which suggests that humans may perform the biotransformation of ABP more efficiently than mice. Second, at a similar bladder tumor incidence, mouse bladder contained adduct concentrations that were much higher than those observed in human bladder; for example, at a 2.6% tumor incidence, mouse bladder contained an average of 55.5 fmol dG-C8 ABP/microgram DNA (1850 adducts/10(8) nucleotides), while bladders from Caucasian male smokers contained an average of 0.036 fmol dG-C8-ABP/microgram DNA (1.2 adducts/10(8) nucleotides). This suggests that factors other than ABP-DNA adducts, such as adducts of other carcinogens, the influence of promoters, and synergistic effects of all of these factors contribute substantially to smoking related bladder cancer in humans. PMID- 9202756 TI - Exposure assessment of heterocyclic amines (HCAs) in epidemiologic studies. AB - The carcinogenic potential of heterocyclic amines (HCAs) in humans has yet to be established. Epidemiologic studies of colon cancer using crude surrogates for HCAs exposure (e.g., doneness of meat) have produced inconsistent results. To improve exposure assessment of HCAs, we have developed a database of HCA concentrations in commonly consumed meat items cooked by various techniques and degrees of doneness. HCA type and level are dependent on multiple factors, including type of meat (e.g., steak, chicken, bacon), cooking technique (with substantial variability present even within high temperature cooking methods), place of preparation (e.g., home, restaurant, or 'fast-food' restaurant), as well as the degree of doneness and surface browning/charring. We have developed a questionnaire with meat photographs linked to this database, which we are using in a variety of case-control and cohort studies of cancer etiology. In addition, we have carried out a metabolic study of HCA exposure among 66 subjects to identify biomarkers of HCA exposure which may be useful in epidemiologic studies. These studies should help clarify the role of HCAs in human carcinogenesis, and eventually allow an estimation of the cancer burden in the population attributable to these compounds. PMID- 9202755 TI - DNA adducts of heterocyclic amines: formation, removal and inhibition by dietary components. AB - The dietary mutagens 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) and 2-amino-1 methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) are carcinogenic in rodents. In F344 rats PhIP induces mammary tumors in females and colon tumors in males, while IQ induces tumors principally in the liver, Zymbal gland and intestines. In CDF1 mice, IQ induces liver, lung and forestomach tumors. We have evaluated the dynamics of formation, removal and inhibition of PhIP- and IQ-DNA adducts in these rodents. After bolus doses (50 mg/kg, by gavage) of IQ or PhIP, both IQ- and PhIP-DNA adducts were removed rapidly from both target and nontarget organs, while after 3-4 weeks of feeding IQ or PhIP (0.01-0.04%) adduct removal was much slower. Gavaging of male F344 rats with PhIP (0.1-1000 micrograms/kg/day) for 23 days resulted in accumulation of PhIP-DNA adducts in various organs, but adducts were detectable only at 100 or 1000 micrograms/kg/day. Urinary excretion of unchanged PhIP was a constant proportion (1.6-2.1%) of the daily dose over the entire dose range and was independent of duration of exposure. When weanling female F344 rats were exposed to dietary PhIP (0.01-0.04%) for 1-4 weeks, the presence of either conjugated linoleic acid (CLA; 0.1-1.0%) or indole-3-carbinol (13C; 0.1%) in the diet inhibited PhIP-DNA adduct formation (58-99%) in various organs, including the mammary gland and the colon. Similarly, the inclusion of 0.075% 4-ipomeanol (IPO) in the diet of male CDF1 mice exposed for 3 weeks to dietary IQ (0.01%) resulted in inhibition of IQ-DNA adduct formation (30-59%) in the target organs (liver, lungs, stomach) but not in a number of other organs. It is concluded that (1) the rate of PhIP- and IQ-DNA adduct removal depends on the dose and frequency of administration, (2) urinary PhIP may be a good biomarker of recent PhIP exposure and (3) CLA, I3C and IPO are potential chemopreventive agents against PhIP- or IQ-induced tumors in rodents. PMID- 9202757 TI - Metabolism of food-derived heterocyclic amines in nonhuman primates. AB - During the cooking of meats, several highly mutagenic heterocyclic amines (HCAs) are produced. Three HCAs, IQ, MeIQx, and PhIP have been under study for carcinogenicity in cynomolgus monkeys, and to date, IQ has been shown to be a potent hepatocarcinogen. Concomitantly, the metabolic processing of these HCAs has been examined. Metabolism studies show that the potent hepatocarcinogenicity of IQ is associated with the in vivo metabolic activation of IQ via N hydroxylation and the formation of DNA adducts. In monkeys undergoing carcinogen bioassay with IQ, N-hydroxylation was confirmed by the presence of the N-hydroxy N-glucuronide conjugate of IQ in urine. The N-hydroxylation of IQ appears to be carried out largely by hepatic CYP3A4 and/or CYP2C9/10, and not by CYP1A2, an isoform not expressed in liver of this species. Notably MeIQx is poorly activated in cynomolgus monkeys and lacks the potency of IQ to induce hepatocellular carcinoma after a 5-year dosing period. The poor activation of MeIQx appears to be due to the lack of constitutive expression of CYP1A2 and an inability of other cytochromes P450, such as CYP3A4 and CYP2C9/10, to N-hydroxylate the quinoxalines. MeIQx is detoxified in monkeys largely by conjugation with glucuronide at the N-1 position. Although the carcinogenicity of PhIP is not yet known, the metabolic data suggest that PhIP will be carcinogenic in this species. PhIP is metabolically activated in vivo in monkeys by N-hydroxylation, as discerned by the presence of the N-hydroxy-N-glucuronide conjugate in urine, bile, and plasma. PhIP also produces DNA adducts that are widely distributed in tissues. The results from these studies support the importance of N-hydroxylation in the carcinogenicity of HCAs in nonhuman primates and by analogy, the importance of this metabolic activation step in the possible carcinogenicity of dietary HCAs in humans. PMID- 9202758 TI - Overview of carcinogenic heterocyclic amines. PMID- 9202759 TI - Interaction of c-myc with transforming growth factor alpha and hepatocyte growth factor in hepatocarcinogenesis. AB - Double transgenic mice bearing fusion genes consisting of mouse albumin enhancer/promoter-mouse c-myc cDNA and mouse metallothionein 1 promoter-human TGF alpha cDNA were generated to investigate the interaction of these genes in hepatic oncogenesis and to provide a general paradigm for characterizing the interaction of nuclear oncogenes and growth factors in tumorigenesis. Coexpression of c-myc and TGF-alpha as transgenes in the mouse liver resulted in a tremendous acceleration of neoplastic development in this organ as compared to expression of either of these transgenes alone. The two distinct cellular reactions that occurred in the liver of the double transgenic mice prior to the appearance of liver tumors were dysplastic and apoptotic changes in the existing hepatocytes followed by emergence of multiple focal lesions composed of both hyperplastic and dysplastic cell populations. These observations suggest that the interaction of c-myc and TGF-alpha, during development of hepatic neoplasia contributes to the selection and expansion of the preneoplastic cell populations which consequently increases the probability of malignant conversion. These studies have now been extended to examine the interaction of hepatocyte growth factor (HGF) with c-myc during hepatocarcinogenesis in the transgenic mouse model. While sustained overexpression of c-myc in the liver leads to cancer, coexpression of HGF and c-myc in the liver delayed the appearance of preneoplastic lesions and prevented malignant conversion. Similarly, tumor promotion by phenobarbital was completely inhibited in the c-myc/HGF double transgenic mice whereas phenobarbital was an effective tumor promoter in the c myc single transgenic mice. The results indicate that HGF may function as a tumor suppressor during early stages of liver carcinogenesis, and suggest the possibility of therapeutic application for this cytokine. Furthermore, we show for the first time that interaction of c-myc with HGF or TGF-alpha results in profoundly different outcomes of the neoplastic process in the liver. PMID- 9202760 TI - Formation and persistence of DNA adducts of 2-amino-3-methylimidazo[4,5 f]quinoline in the rat and nonhuman primates. AB - The formation and persistence of the two principal DNA adducts of the food derived carcinogen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) have been investigated in rats and nonhuman primates. DNA adduct formation in the liver of male Fischer-344 rats occurred in a dose-dependent manner (0.01-20 mg/kg) where N (deoxyguanosin-8-yl)-2-amino-3-methylimidazo[4,5-f]quinoline (dG-C8-IQ) and 5 (deoxyguanosin-N2-yl)-2-amino-3-methylimidazo[4,5-f] quinoline (dG-N2-IQ) accounted for approximately 60-80% and 20-40%, respectively, of the total adducts observed by 32P postlabeling. Similar DNA adduct profiles were observed in kidney and colorectal tissue of rats given a single oral dose of IQ (20 mg/kg) which, when given chronically to this species, results in tumorigenesis in liver and colorectum, but not in kidney. dG-C8-IQ was removed more rapidly than dG-N2-IQ from liver and kidney, but removal of both adducts from the colorectum closely followed cell replication. Similar DNA adduct profiles were observed in liver and extrahepatic tissues of nonhuman primates following a single dose of IQ (20 mg/kg). In chronically treated monkeys undergoing carcinogen bioassay, there was a sharp increase in the contribution of dG-N2-IQ to total DNA adducts in all slowly dividing tissues. There was no preferential accumulation of dG-N2-IQ in the colon, a tissue with a high rate of cell division, and dG-C8-IQ remained the predominant lesion. These findings point to a preferential removal of the dG-C8 IQ adduct by enzyme repair system(s) in slowly dividing tissues in both rats and nonhuman primates. PMID- 9202761 TI - MeIQx-DNA adduct formation in rodent and human tissues at low doses. AB - Heterocyclic amines, such as 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), are mutagenic/carcinogenic compounds formed during the cooking of protein-rich foods. Human exposure to MeIQx has been estimated to range from ng/person/day to a few microgram/person/day. In contrast, animal studies have been conducted at doses in excess of 10 mg/kg/day. In order to determine the relevance of high-dose animal data for human exposure, the dose-response curves for [14C]-MeIQx have been determined in rodents at low doses under both single dose and chronic dosing regimens using the high sensitivity of accelerator mass spectrometry (AMS). To make a direct species comparison, rodent and human colonic MeIQx-DNA adduct levels have been compared following oral administration of [14C] MeIQx. The results of these studies show: (1) total MeIQx levels are highest in the liver > kidney > pancreas > intestine > blood; (2) MeIQx levels in the liver plateau after 7 days of chronic feeding; (3) hepatic MeIQx-DNA adducts begin to plateau after 2-4 weeks and reach steady-state levels between 4 and 12 weeks on chronic exposures; (4) hepatic DNA adducts generally increase as a linear function of administered dose for a single-dose exposure and as a power function for chronic feeding over a dose range spanning 4 orders of magnitude; (5) human colon DNA adduct levels are approximately 10 times greater than in rodents at the same dose and time point following exposure; and (6) > or = 90% of the MeIQx-DNA adduct in both rodent and human colon appears to be the dG-C8-MeIQx adduct. These studies show that MeIQx is readily available to the tissues for both humans and rodents and that adduct levels are generally linear with administered dose except at high chronic doses where adduct levels begin to plateau slightly. This plateau indicates that linear extrapolation from high-dose studies probably underestimates the amount of DNA damage present in the tissues following low dose. Further, if adducts represent the biologically effective dose, these data show that human colon may be as sensitive to the genotoxic effects of MeIQx as rat liver. The significance of these endpoints to tumor response remains to be determined. PMID- 9202762 TI - Human exposure to mutagenic/carcinogenic heterocyclic amines and comutagenic beta carbolines. AB - Various kinds of mutagenic and carcinogenic heterocyclic amines (HCAs) are produced by heating protein-rich foods, such as meat and fish. To evaluate the risk of these HCAs in terms of human cancer development, exposure levels must be measured. We therefore analyzed their amounts in various kinds of cooked foods and in urine samples of healthy volunteers living in Tokyo. Based on the obtained quantitative data, daily exposure levels to 2-amino-3,8-dimethylimidazo[4,5 f]quinoxaline (MeIQx) and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) were calculated to be 0.3-3.9 and 0.005-0.3 microgram per person, respectively. Moreover, human DNA samples were analyzed with the 32P-postlabeling method, and colon, rectum and kidney tissues were found to contain an adduct spot corresponding to the standard 5'-pdG-C8-MeIQx by TLC and HPLC, at levels of 14, 18 and 1.8 per 10(10) nucleotides, respectively. The beta-carboline compound, norharman, is produced by heating L-tryptophan, and is known to be present in cooked foods and in cigarette smoke at higher levels than mutagenic and carcinogenic HCAs. While norharman is not itself mutagenic to Salmonella, it does become mutagenic to S. typhimurium TA98 with S9 mix in the presence of non mutagenic aromatic amines like aniline and o-toluidine. When we examined whether DNA adducts are formed in the DNA of S. typhimurium TA98 by treatment with norharman and aromatic amines using 32P-postlabeling analysis, DNA adduct formation by norharman with aromatic amines was found to be related to the appearance of mutagenicity by norharman with aromatic amines. PMID- 9202763 TI - A perspective on the history and significance of carcinogenic and mutagenic N substituted aryl compounds in human health. PMID- 9202764 TI - Arylamine activating sulfotransferase in liver. AB - Carcinogenic N-hydroxy-arylamines and -arylamides undergo metabolic activation by several enzymes in mammals to cause the DNA damage. Cytosolic sulfotransferases in rat and human livers, which belong to the ST1 (SULT1) family, have been studied to assess their properties to mediate the metabolic activation. A human orthologue of rat ST1C1 from, which catalyzes sulfation of N-hydroxy-2 acetylaminofluorene, was screened in a EMBL genomic library with ST1C1 cDNA [Nagata, K., S. Ozawa, M. Miyata, M. Shimada, D.-W. Gong, Y. Yamazoe and R. Kato (1993) J. Biol. Chem., 268, 24720-24725]. Sequencing of the hybridized clones indicate that at least 3'-terminal region of human ST1C1 orthologue contains sequence highly homologus to rat ST1C1 at both nucleotide and deduced amino acid levels. The experiments using anti-rat ST1C1 antibody and nucleotide probes for human ST1C1 showed no detectable band in Western blots and an mRNA-detecting method with polymerase chain reaction, respectively, in human liver samples. PMID- 9202765 TI - Prognostic factors in severely head injured adult patients with epidural haematoma's. AB - A medline search back to 1975 was undertaken to identify relevant papers published on epidural haematomas. The search was restricted, whenever possible, to adult age and to comatose patients. Forty four relevant reports were identified. Only 4 papers reported results on multivariate analysis. In terms of prognosis, the following parameters were found to be significant: age, time from injury to treatment, immediate coma or lucid interval, presence of pupillary abnormalities, GCS/motor score on admission. CT findings (haematoma volume, degree of midline shift, presence of signs of active haematoma bleeding, associated intradural lesion) and post-operative ICP. To compare different casistics we need more informations about patients's outcome in the referral area of the neurosurgical centers, about the number of direct admissions and about the number of patients showing clinical deterioration. PMID- 9202766 TI - Prognostic factors in severely head injured adult patients with acute subdural haematoma's. AB - A medline search back to 1975 was undertaken to identify relevant papers published on subdural haematomas. The search was restricted, whenever possible, to adult age and comatose patients. Forty relevant reports were identified. Only 3 papers reported results on multivariate analysis. In terms of prognosis, the following parameters were found to be significant: age, time from injury to treatment, presence of pupillary abnormalities, GCS/motor score on admission, immediate coma or lucid interval, CT findings (haematoma volume, degree of midline shift, associated intradural lesion, compression of basal cisterns), post operative ICP and the type of surgery. Improving the outcome of patients with acute subdural haematoma's is a difficult task. A small subpopulation of patients may have a benign course without surgical haematoma evacuation, but all comatose patients with an acute subdural haematoma should be treated in Centers where neurosurgical facilities and appropriate monitoring are available. PMID- 9202767 TI - EBIC-guidelines for management of severe head injury in adults. European Brain Injury Consortium. AB - Guidelines for the management of severe head injury in adults as evolved by the European Brain Injury Consortium are presented and discussed. The importance of preventing and treating secondary insults is emphasized and the principles on which treatment is based are reviewed. Guidelines presented are of a pragmatic nature, based on consensus and expert opinion, covering the treatment from accident site to intensive care unit. Specific aspects pertaining to the conduct of clinical trials in head injury are highlighted. The adopted approach is further discussed in relation to other approaches to the development of guidelines, such as evidence based analysis. PMID- 9202768 TI - Post-traumatic bilateral diffuse cerebral swelling. AB - This study confirms that bilateral diffuse cerebral swelling with or without parenchymal haemorrhages (< 15 cc) is a more common occurrence in the paediatric patients with severe head injury as compared with adults, since the analysed sample represented 42.55% and 20.43% of all paediatric and adult patients with severe head injury recorded in our clinic at the time of the study, respectively. The incidence of patients with diffuse cerebral swelling without parenchymal haemorrhages was found to be 27.65% of paediatric patients and 5.37% of adult patients with severe head injury. Secondary neurological deterioration occurred only in 5 (12.5%) paediatric patients and in 4 (10.5%) adult patients with diffuse cerebral swelling and was not to be found associated with parenchymal haemorrhages. A better outcome was seen in paediatric patients. Mortality rates were 12.5% in paediatric patients and 34.21% in adult patients. Our data also suggest that the mortality rate between paediatric and adult patients with diffuse cerebral swelling without parenchymal haemorrhages was similar (15.38% and 20% in paediatric and adult group, respectively), while the adult patients with diffuse cerebral swelling associated with small intraparenchymal haemorrhages have a worse prognosis than paediatric patients. PMID- 9202769 TI - The Kluver-Bucy syndrome. AB - Evolution of psychological disorders following head injury including memory and other cognitive disorders are common. The best known are psychiatric disturbances of various kinds after lesions of the frontal lobes. Cognitive, behavioural and emotional disorders are not usually seen in patients with bilateral temporal lesions. In our Department of Neurotraumatology we have observed 4 patients with posttraumatic lesions localized bitemporally. They developed Kluver-Bucy syndrome rarity in human pathology-combined with three or more of the following symptoms and signs: increased oral activity, hypersexuality, hypermetamorphosis, memory disorders, placidity, loss of people recognition, bulimia. Several symptoms responded dramatically to carbamazepine. We conclude that it may be a useful agent in the treatment of this unusual syndrome. PMID- 9202770 TI - Dural arteriovenous fistulas including the transverse and sigmoid sinuses: results of treatment in 30 cases. AB - We report about the treatment and outcome of 30 patients with dural arteriovenous fistulas including the transverse and sigmoid sinuses treated between 1986 and 1995. All patients underwent panangiography for definitive diagnosis. The dAVF were supplied by the external carotid artery system alone (14 patients), both external and internal carotid systems (10 patients) or both anterior and posterior circulation (6 patients). Depending on the venous drainage the fistulas were classified following a modification of Djindjian's description with 18 patients revealing Type I (main sinus with antegrade flow), 5 Type II a (main sinus with reflux into the contralateral sinus). 5 Type II b (cortical veins), 1 Type II a+b (both) and 1 of Type III (direct cortical drainage). Bruit, pulsatile tinnitus and headaches were the most common symptoms. 6 patients presented with intracranial haemorrhage, 4 with progressive neurological deficit or seizures and 3 with dementia. Arterial embolization was performed in all cases except one, where a transvenous approach for balloon occlusion of the transverse sinus was performed. 21 patients were treated by single or repeated embolization alone. Only in 9/21 cases did arterial embolization result in complete occlusion of the fistula. In 12/21 patients incomplete occlusion was achieved. Following embolization 8 patients underwent additional surgery including coagulation of the feeding arteries and arterialized veins, sinus resection and reconstruction of the sinus. Overall, 18 patients were cured, 11 improved and 1 patient was unchanged. There was a total number of 5 complications including transient stroke, transient facial nerve palsy, and a small necrotic skin area following embolization. Venous infarction of the occipital lobe was induced by transvenous occlusion and surgical resection of the transverse sinus in one patient each, respectively. From our results we conclude that the endovascular therapy alone is the treatment of choice in case of Type I fistulas. In dAVF of Type II and III repeated endovascular treatment seems not to be sufficient and additional surgery is necessary. PMID- 9202771 TI - Platelet derived growth factor and subarachnoid haemorrhage: a study on cisternal cerebrospinal fluid. AB - Platelet derived growth factor (PDGF) was identified as a powerful mitogenic growth factor which is released from activated platelets and has a marked activity as vasoconstrictor agent. In the present study we have measured cisternal cerebrospinal fluid (CSF) levels of PDGF in 72 patients operated on for intracranial aneurysm in order to verify whether it might be related to the clinical aspects of SAH with special regard to symptomatic vasospasm. CSF samples were obtained at surgery by cisternal puncture of the subarachnoid cistern the nearest to the aneurysm before aneurysm isolation and exclusion. The specimen were frozen in liquid nitrogen and stored at -80 degrees C until analysis. PDGF was measured using a commercially available reagent. Values are expressed as pg/ml of CSF. In 18 cases no radiological and clinical signs of SAH were detected and the mean cisternal CSF level of PDGF was 885.0 +/- 104.5 pg/ml; 20 patients were operated on between day 1 and 3 from the last SAH episode: mean cisternal CSF level of PDGF was 1917.5 +/- 459.4 pg/ml. In 34 patients treated with delayed surgery protocol, mean cisternal CSF level of PDGF was 995.3 +/- 73.8 pg/ml. Statistical analysis showed significant differences between groups (P: 0.011). In the subgroup of patients operated on within day 3 after SAH, 6 presented vasospasm and had mean cisternal CSF PDGF level which was significantly higher (P < 0.01) than in 14 patients without vasospasm. In the delayed "surgical" patients there was no significant difference in cisternal CSF levels of PDGF considering the occurrence of vasospasm. The results of the present study suggest that (a) after SAH there is a significant release of PDGF early after SAH and (b) higher levels of PDGF found in cisternal CSF of patients operated on within 72 hours after SAH may be predictive of symptomatic vasospasm. PMID- 9202772 TI - Management strategies for posterior cerebral artery aneurysms: a proposed new surgical classification. AB - In a period of 10 years fifteen patients bearing sixteen aneurysms arising at the posterior cerebral artery were operated at our institution. Based on the approaches selected for each location a division of the posterior cerebral artery into three surgical segments is proposed. The first segment (S1), or anterior extends from the basilar artery bifurcation to the point where the artery reaches the level of the most lateral edge of the cerebral peduncle, the second segment (S2), or middle extends from the posterior limit of S1 to a point located just before the most medial extent of the artery in the quadrigeminal cistern (collicular point), and the third segment (S3), or posterior corresponds to the collicular point and to the portions of the posterior cerebral artery distal to it. Utilizing the concept of surgical segments all aneurysms in our series were satisfactorily exposed. Those arising at the S1 segment (8 cases) were operated either through a pterional or a pretemporal approaches; those from the S2 segment (6 cases) were operated either via the subtemporal or the subtemporal transventricular routes; and that arising from the S3 segment (1 case) was managed through the occipital interhemispheric approach. Among the aneurysms eleven were small, one was large, and four were large or giant. Ten of these aneurysms were surgically clipped, two coagulated, three treated by trapping and in one case the aneurysm was resected and the posterior cerebral artery was reconstructed by a termino-terminal anastomosis. The surgical results were considered good in all cases but one, where the patient died due to clinical complications three months after surgery. It is our belief that the use of this classification can provide the means to best select the most appropriate surgical approach to treat aneurysms arising at the posterior cerebral artery. PMID- 9202773 TI - Superior sagittal sinus thrombosis treated with combined local thrombolytic and systemic anticoagulation therapy. AB - We recently encountered a patient with thrombosis of the superior sagittal sinus of an idiopathic cause. The patient was treated initially with combined local thrombolytic therapy through the burr hole over the superior sagittal sinus and systemic anticoagulant therapy. Continuous ventricular drainage and hyperbaric oxygenation therapy were used to control the increased intracranial pressure. The superior sagittal sinus was successfully recanalized. Whereas the patient suffered a complication with subacute subdural haematoma, he was successfully treated with the combination of these therapies. The rationale and approach are discussed. PMID- 9202774 TI - Thalamic gliomas: a clinicopathologic analysis of 20 cases with reference to patient age. AB - Twenty patients (M 11, F9; ranging from 1-77 years old) with histologically proven glial tumours in the thalamic region, treated from 1979 until 1994 at Kyushu University Hospital were retrospectively reviewed and analysed in order to elucidate their clinical and neuropathological characteristics. The initial common clinical manifestations were those of increased intracranial pressure or motor weakness. The histological diagnosis of the tumour was pilocytic astrocytoma in 2 patients, fibrillary astrocytoma in 7, anaplastic astrocytoma in 7, and glioblastoma multiforme in 4. The initial treatment was surgery alone in 4 patients, surgery followed by radiation therapy in 5, surgery followed by radiation therapy and chemotherapy in 9, and conventional radiation therapy alone in 2 patients. The 3-year overall actuarial survival rate for all patients was 20% but was related to both the histological type and the age of the patients: As a result, the rate was 44% for patients with low-grade astrocytoma compared to 0% for those with high-grade astrocytoma. While 5 out of 11 patients under the age of 25 years at their initial presentation have survived for from 2-16 years after the diagnosis, all patients presenting after the age of 25 years died within 3 years after treatment. Thalamic glial tumours are not a homogeneous group of tumours in terms of clinical behaviour and histopathological features, and the poor overall results, especially in adult tumours, thus emphasise the need for continued research in the treatment of these tumours. PMID- 9202775 TI - Endoscopic transsphenoidal resection of a large chordoma in the posterior fossa. AB - Encouraged by an experience with endoscopic transsphenoidal pituitary surgery, an endoscopic transsphenoidal technique was applied in a patient with a large chordoma in the posterior fossa. The patient was a 40-year-old man with a two year history of progressive ataxia, a memory disorder and emotional instability. A magnetic resonance (MR) scan of the brain revealed a midline posterior fossa mass measuring 4 cm in diameter located between the clivus and the brainstem. The basilar artery and its bifurcation were encased by the tumor and the brainstem was also distorted by the tumor. Obstructive hydrocephalus was treated previously with a ventriculoperitoneal shunt and fractionated external beam radiation treatment was given without histological diagnosis at another hospital. Subtotal resection of the tumor was achieved utilizing an endoscopic transphenoidal technique through the patient's nostril. The portion of the tumor located behind the basilar artery was not resected in order to protect the brainstem perforating arteries. The patient showed dramatic improvement of his symptoms postoperatively. Residual tumor located behind the basilar artery was treated by stereotactic gamma-knife surgery. This is the first reported case of a large posterior fossa chordoma being treated by an endoscopic transsphenoidal technique. PMID- 9202776 TI - Progesterone receptors in arachnoid cysts. An immunocytochemical study in 2 cases. AB - We report 2 cases of arachnoid cysts, one with a retrocerebellar and the other with a left temporal localization, in which immunohistochemical studies had been conducted. The results of the immunohistochemistry on the presence of carcino embryonic antigen (CEA) and glial fibrillary acidic protein (GFAP), and of the scanning- and transmission electron microscopy revealed the cyst lining to be identical to subdural neurothelium. Progesterone receptors were found in the nuclei of cells lining the cyst, which also suggests the similarity of the cyst lining to arachnoid granulations and meningiomas as derivatives of subdural neurothelium, which also possess progesterone receptors. PMID- 9202777 TI - Control of complications in the midfrontobasal approach. AB - Many surgical techniques are employed for access to midline lesions of the anterior and middle fossa. A convenient one is the midfrontobasal approach. For various indications this surgical procedure was performed in 105 patients. Its benefits are outlined and a method for dealing with the two main drawbacks, postoperative loss of the sense of smell and the presumed risk of infection related to the inevitable opening of the frontal sinus, are described. In 83% of the cases the authors were able to anatomically retain the olfactory nerves, with preservation of the sense of smell in at least 65% of them. The frontal sinus was opened in 76% of the cases. The incidence of infection in the simple and quick integral reconstruction appeared 1%, equal to the rate of infection in the classical reduction of the frontal sinus by a periosteal flap. PMID- 9202778 TI - Circular stereotactic callosotomy: a preliminary report. Technical note. AB - The authors propose a new method for performing stereotactic callosotomy, which they have named circular callosotomy. The operating device is the original Riechert-Mundinger's string electrode, which can be protruded through a side window and by rotating the probe it is possible to cut the commisural pathways to the extent required. The anatomical results of the operation can be checked using MRI scanning. PMID- 9202779 TI - The clinoidal space: anatomical review and surgical implications. AB - This study describes the anatomy of the clinoidal space exposed after the anterior clinoid process (ACP) is removed. Five cadaver heads injected with colored latex in the arterial and venous systems were used. Each was cut in half to provide ten specimens for inspection. The bone that covered the medial side of the cavernous and clinoidal internal carotid artery (ICA) was removed. The ACP was removed and its dural layers were preserved. The removal of the ACP establishes an area called the clinoidal space. In this space, the clinoidal ICA is exposed. This space is delimited by two dural rings that anchors the clinoidal ICA. Most of the clinoidal space is located anterolateral to the artery where the ACP is found, but there is a small triangular space posterior to the artery and another space anteromedial to it. The clinoidal ICA is completely encased by connective tissue in this space. The clinoidal space is extracavernous, therefore, bleeding occurs only if the connective tissue layer is broken. PMID- 9202780 TI - Morphology and immunohistochemistry of a symptomatic septum pellucidum cavum Vergae cyst in man. AB - The cavum septi pellucidi (CSP) and cavum Vergae (CV) are persistent, primitive, or acquired, midline structures of adult human brain. It is customary to distinguish between the non-communicating and the communicating cava, depending on whether the cavum communicates with the cerebral ventricular system or not. Only a few cases of symptomatic non-communicating cava, called septum pellucidum and cavum Vergae cysts, have been described in the literature. In this study, the authors describe the morphological, histological and histo-immunological characteristics of an additional case of septum pellucidum-cavum Vergae cyst in a forty-year-old man who died the day following a ventriculo-peritoneal shunt. We have found a communication between the CSP and the leptomeningeal space of the anterior interhemispheric fissure, in the absence of subarachnoid haemorrhage. The authors discuss the origin of the intracystic fluid and the classification of the CSP. PMID- 9202781 TI - Fatal complication of the percutaneous radiofrequency trigeminal rhizotomy. PMID- 9202782 TI - Cerebral revasculization in aortitis with occluded bilateral common carotid and left vertebral arteries. PMID- 9202784 TI - Ectopic craniopharyngioma: presentation of a case arising from the corpus callosum. PMID- 9202783 TI - Suboccipital tangential gunshot wound. PMID- 9202785 TI - Short-term measurements of linear growth in early life: infant knemometry. PMID- 9202786 TI - Preventive strategies for young children with wheezing bronchitis. PMID- 9202787 TI - Experimental milk formulae with reduced protein content and desialylated milk proteins: influence on the faecal flora and the growth of term newborn infants. AB - We have assessed the growth, tolerance and the faecal flora composition in healthy infants on different feeding regimens. Four groups of infants were fed exclusively on mother's milk, a standard formula and two experimental formulae. The first experimental formula consisted of a milk with a reduced protein content (1.2 g/100 ml), the second in a formula with the same protein content and with milk proteins desialylated by mild acid hydrolysis. The aim of the study was to test whether lowering the protein content and/or modifying the proteins by desialylation would favour the development of a bifidus flora. A bifidus flora was detected in 60% of breastfed infants at 1 month of life. All formulae employed during the study failed to induce a prevalence of colonization with bifidobacteria at 1 month of age. The two experimental milk formulae were well tolerated, but the infant growth rate was slightly lower as compared to the breastfed infants and the infants fed the standard formula. The presence in milk formulae of pre-digested and desialylated proteins can offer some advantages in term of digestibility and mimic a physiological intestinal mechanism of the infant. PMID- 9202788 TI - Breastfeeding and catch-up growth in infants born small for gestational age. AB - Postnatal growth was prospectively measured from birth to 1 y in 54 term infants born small for gestational age (SGA), fed either breast milk or a standard term infant formula. Breastfeeding was associated with a 0.36 and 0.64 standard deviation (SD) increase in weight at 2 weeks and 3 months of age, respectively, which persisted beyond the breastfeeding period (0.64 SD at 1 y). Breastfed infants also showed greater catch-up growth in head circumference [SD score (SDS) 0.53 higher at 3 months], and greater body length gain (SDS 0.68 higher at 6 months). This increased growth was independent of potentially confounding obstetric, social and demographic factors. Our findings suggest that breastfeeding may promote faster growth in infants compromised by poor growth in utero. SGA infants may be programmed for a number of adverse outcomes; the possibility that such events are altered by choice of postnatal diet is a key issue for future research. PMID- 9202789 TI - Bone mineral acquisition in low calcium intake children following the withdrawal of calcium supplement. AB - Our recent 18-month calcium supplementation trial demonstrated a significant increase in radial bone mineral mass in 7-year-old children with calcium intake approximately 300 mg/day (Am J Clin Nutr 1994; 60: 744-50). The persistence of higher bone mass after cessation of calcium supplementation is unknown. This is a follow-up study to investigate the lasting effect of calcium supplementation on bone acquisition. Subjects were 159 Chinese children aged 8.7 years. Distal one third radial bone mineral content (BMC) and bone width (BW) were measured by single-photon absorptiometry. After 12 months, the significant difference in mean +/- SD percentage radial BMC disappeared between the study and control groups (7.34 +/- 6.77% vs 8.67 +/- 6.46%, p > 0.05). Dietary calcium intakes were similar between the groups. During the supplementation phase, the study group had 17.9% greater BMC gain than that of controls. In the follow-up phase, however, the study group had 16.1% less BMC gain than that of controls. It appears that an increased acquisition rate during the supplementation phase was almost balanced by a reduced acquisition rate during follow-up phase. Moreover, throughout the entire 30-month period, the overall BMC acquisition rates of the study and control groups were 25% and 23.8%, respectively. Hence, the overall acquisition rate of the study group was only 5% higher than that of controls. Therefore, the effect of calcium supplementation on bone mineral gain appears to reflect a transient reduction in bone turnover rate. Longer-term calcium trials are necessary to confirm whether a sustainable higher calcium intake throughout childhood will enhance peak bone mass. PMID- 9202790 TI - Randomized trial of high nutrient density formula versus standard formula in chronic lung disease. AB - To test the hypothesis that the nutrient intake and growth of infants with chronic lung disease would be improved by providing nutrients in more concentrated form, and that the lower volume would improve respiratory status, 60 preterm infants requiring supplemental oxygen at 28 days of age were randomly assigned a low-density (24 kcal/oz) formula fed at 180 ml/kg/day, or a high density (30 kcal/oz) formula at 145 ml/kg/day. There was no difference in growth, respiratory outcome, oedema or diuretic requirement between dietary groups. Infants fed the high nutrient density formula had significantly greater total energy (143 vs 134 kcal/kg/day) and protein (3.9 vs 3.6 g/kg/day) intakes, largely because the low-density formula group did not achieve the designated 180 ml/kg/day. In conclusion, use of a high-density formula in infants with chronic lung disease did not improve growth or respiratory outcome, despite a small increase in total energy and protein intake. PMID- 9202791 TI - Comparison of total alkaline phosphatase and three assays for bone-specific alkaline phosphatase in childhood and adolescence. AB - We compared serum levels of total alkaline phosphatase (TAP) and bone-specific alkaline phosphatase (BAP) as determined by three different assays (lectin affinity electrophoresis, immunoradiometric assay, enzyme-linked immunosorbent assay) in subjects aged 5-20 years suffering from X-linked hypophosphatemic rickets (n = 14), chronic renal failure (n = 10) and chronic cholestatic liver disease (n = 16). Results were compared to controls of the same age and were expressed as standard deviation scores (SDS). TAP correlated significantly with BAP (r > 0.9 for each assay; p < 0.001) in controls. In children with cholestatic diseases, TAP (median SDS + 2.0) was elevated, but BAP, as measured by the electrophoretic assay, was within the reference range for most patients (median SDS: -0.4; p = 0.003 for the difference between the median SDS of TAP and BAP). In contrast, results for BAP as determined by the two immunoassays were not significantly different from TAP in any of the three patient groups (p > 0.05 in each group for both assays). In this study, the two immunoassays did not have a detectable advantage over lectin affinity electrophoresis in the determination of BAP. PMID- 9202792 TI - Iron status of children with short stature during accelerated growth due to growth hormone treatment. AB - We determined the influence of human growth hormone (hGH) treatment on blood soluble transferrin receptor (sTfR) in 35 children with short stature. Whereas the serum concentration of ferritin decreased from 29.6 micrograms/l to 19.7 micrograms/l, and that of transferrin increased from 2.9 g/l to 3.2 g/l during 6 months (p < 0.001), only a minimum rise in the sTfR concentration was observed (7.12 +/- 0.20 mg/l vs 7.51 +/- 0.19 mg/l, p = 0.025). The prevalence of anaemia or microcytosis did not increase. Most of the changes in serum ferritin and transferrin concentrations occurred during the first week. The study demonstrates that rapid body growth per se does not affect the sTfR concentration, but it may affect the serum transferrin and ferritin concentrations. Alternatively, GH may have a specific effect on serum ferritin and transferrin concentrations. PMID- 9202793 TI - Vitamin D metabolites (25-hydroxyvitamin D, 24,25-dihydroxyvitamin D and 1,25 dihydroxyvitamin D) and osteocalcin in beta-thalassaemia. AB - Serum levels of the vitamin D metabolites 25-hydroxyvitamin D, 24,25 dihydroxyvitamin D, and 1,25-dihydroxyvitamin D, and of osteocalcin, C-terminal parathyroid hormone and other biochemical indices related to bone metabolism, were determined in two groups of patients with beta-thalassaemia aged 5-10 years (summer 7.8 +/- 0.4 years, mean +/- SEM, and winter 7.7 +/- 0.4 years, group A, n = 15) and 11-23 years (16.6 +/- 0.9 and 15.7 +/- 0.9 years in summer and winter, respectively, group B, n = 22). Emphasis was given to populations of school and adolescent ages and to the seasons of summer and winter when vitamin D status demonstrates the widest extremes. The mean serum levels of 25-hydroxyvitamin D in patients aged 5-10 years did not differ from those of controls during both seasons studied. In contrast, in the age group 11-23 years these levels were found to be lower in patients than in controls both in winter (10.6 +/- 0.9 ng/ml vs 15.0 +/- 2.0 ng/ml, p < 0.05) and summer (20.2 +/- 2.1 ng/ml vs 27.1 +/- 2.0 ng/ml, p < 0.05). The serum concentrations of 24,25-dihydroxyvitamin D were lower in the thalassaemic patients than in controls in both age groups and both seasons. In the patients under 10 years of age the mean values of this metabolite in winter were 1.06 +/- 0.17 ng/ml vs 1.68 +/- 0.20 ng/ml in the respective controls (p < 0.05), and in summer 1.44 +/- 0.11 ng/ml vs 2.35 +/- 0.36 ng/ml in controls (p < 0.05). In the group of patients aged 11-23 years, the mean levels of 24,25-dihydroxyvitamin D were in winter 0.65 +/- 0.12 ng/ml vs 1.12 +/- 0.19 ng/ml (p < 0.05) in controls and in summer 1.34 +/- 0.12 ng/ml vs 1.84 +/- 0.20 ng/ml (p < 0.05). The 1,25-dihydroxyvitamin D concentrations in both thalassaemic patient groups were significantly no different from those in the respective control groups. Serum osteocalcin, C-terminal parathyroid hormone, calcium, inorganic phosphate and alkaline phosphatase levels in the patients studied were not significantly different from those in controls, except for calcium and phosphate in the older group. In the older group of thalassaemic patients, serum calcium was lower than in the controls (2.26 +/- 0.03 vs 2.37 +/- 0.03 mmol/l, p < 0.05) in summer and serum phosphate higher than in the controls in winter (1.47 +/- 0.05 mmol/l vs 1.27 +/- 0.06 mmol/l, p < 0.05). PMID- 9202794 TI - Pre- and postnatal medical care and risk of sudden infant death syndrome. AB - The current study investigated whether sufficient attendance at prenatal and postnatal checks affects the risk of sudden infant death syndrome. A case-control study in the Tyrol enrolled 99 infants with sudden infant death syndrome that occurred between 1984 and 1994, and 136 randomly selected control cases. The risk of sudden infant death syndrome was higher in infants whose mothers attended less than five antenatal health checks than in the group with at least five or more visits (OR 5.1; p < 0.01). Babies who received medical help beyond routine health controls had a lower risk than those who did not (OR 0.32; p < 0.001). These differences persisted when social and demographic variables (mother's age at delivery, educational level, marital status, parity and gestational age) were taken into account. Our study identified inadequate antenatal and postnatal care as a risk indicator for sudden infant death syndrome and as a potential target for further educational work. Clinical recommendations should await the results of further evaluations. PMID- 9202795 TI - Primary infections of human herpesvirus-7 and herpesvirus-6: a comparative, longitudinal study up to 6 years of age. AB - In order to understand the infection status of human herpesvirus 7 (HHV-7) in Taiwan and clarify the serological cross-reactivity between HHV-7 and HHV-6, a longitudinal study was carried out in 52 infants from whom 9 sera were available from birth to 6 years of age. All sera were tested for antibodies against HHV-6 and HHV-7 by indirect immunofluorescence assay. HHV-7 infection was not seen before 6 months of age and gradually emerged thereafter, reaching a cumulative rate of 28.8%, 67.3%, 73.1%, 78.8%, 82.7%, and 86.5% by the ages of 1, 2, 3, 4, 5 and 6 years, respectively. Primary HHV-6 infection induced a reactive HHV-7 antibody in 10/28 (35.7%) cases with a titre of no more than 10. Twenty-eight children (53.8%) were infected by HHV-6 earlier than HHV-7, while eight (15.4%) were infected by HHV-7 earlier than HHV-6. In summary, HHV-7 infected children later than HHV-6. A one-way cross-reaction of HHV-7 serology by HHV-6 was revealed and a titre of 20 is appropriate for defining HHV-7 infection. PMID- 9202796 TI - Increased lipid peroxidation in children with autoimmune diseases. AB - To examine the role of oxidative damage in children and adolescents with autoimmune diseases, we compared blood serum levels of the lipid peroxidation (LPO) products 4-hydroxynonenal (HNE) and malondialdehyde (MDA) in 22 children with systemic lupus erythematosus (SLE), 13 children with focal type of scleroderma, and 21 health controls. In order to study the influence of disease activity in SLE on serum LPO product levels, the SLE group was divided into one group with active disease (n = 11) and one group with non-active disease (n = 11) according to SLEDAI-score, 15.1 and 1.8, respectively. SLE patients with active SLE (146 +/- 14 nmol/l, median 145 nmol/l) have significantly higher HNE levels compared to controls (61 +/- 10 nmol/l, median 52 nmol/l), whereas the MDA serum levels are similar to those of the control group, 1.94 +/- 0.18 mumol/l (median: 2.02 mumol/l) and 1.58 +/- 0.11 mumol/l (median: 1.52 mumol/l), respectively. Children with SCL had HNE and MDA levels similar to the control group. PMID- 9202797 TI - Serum C-reactive protein levels in patients with Kawasaki disease: from the results of nation-wide surveys of Kawasaki disease in Japan. AB - Thirteen nation-wide epidemiological surveys of Kawasaki disease have been carried out successively since 1970 in Japan. In the latest survey, questionnaires on serum C-reactive protein (CRP) levels of the patients were included to clarify whether serum CRP levels could be available for the diagnosis and prediction of prognosis. A questionnaire from and diagnostic guidelines for Kawasaki disease were sent to all paediatric departments of hospitals with 100 or more beds throughout Japan, and information including maximal serum CRP levels was obtained on patients with Kawasaki disease diagnosed during the 2-year period from January 1993 to December 1994. Of the 11458 patients diagnosed during the 2 year period, maximal serum CRP levels were reported in 11040 patients (96.4%). The values of maximal serum CRP were higher in the age groups < 6 months and > 2 years. The mean value and the distribution of serum CRP levels in-suspected cases were lowest among the three diagnostic categories and this difference among diagnostic categories was highly significant in the age groups 6 M-1 Y and 1-2 Y. The proportion of patients with cardiac sequelae increased with serum CRP levels in each age group. The mean value and the distribution of serum CRP levels of the patients with cardiac sequelae was higher than those without it and this difference between cardiac prognoses was outstanding in the age groups 6 M-1 Y and < 6 M. The Receiver/Response Operating Characteristic (ROC) curve for maximal serum CRP levels in Kawasaki disease revealed that accuracy of maximal serum CRP levels for prediction of cardiac sequelae was highest in the age group 6 M-1 Y. A large-scale observation and analysis of serum CRP levels of the patients with Kawasaki disease revealed age-dependent relationships among maximal serum CRP levels, diagnostic categories and prognosis. Serum CRP levels may be helpful for the prediction of prognosis with the consideration of age. PMID- 9202798 TI - Dynamic properties of the aorta and of the foot microcirculation in adolescents with diabetes mellitus. AB - To evaluate vascular function in diabetic subjects, we studied both the stiffness of the abdominal aorta and the foot microvascular reactivity in 22 diabetic adolescents and 18 controls. The aortic stiffness was significantly higher in diabetic females, but not in males, as compared to age- and gender-matched controls (p < 0.05). Foot post-ischaemic hyperaemia was lower in diabetic subjects than in controls (p < 0.05), while postural vasoconstriction did not differ between the groups. The microvascular reactivity did not correlate with the duration of diabetes, but seemed to be influenced by the insulin regimen. The degree of aortic stiffness and the microvascular reactivity of the foot were not significantly interrelated. Loss of aortic elasticity might be a long-term effect of diabetes, whereas microvascular reactivity seems to reflect the current influence of the metabolic state and insulin treatment. PMID- 9202800 TI - Incidence of type I diabetes mellitus in Navarre, Spain (1975-91). AB - The aim of this study was to ascertain the incidence of Type 1 diabetes mellitus in Navarre, an autonomous community in northern Spain. Subjects were patients who presented with diabetes between 1975 and 1991, age range 0-16 years, resident in Navarre at the onset of symptoms. Endocrinologists in outpatient centres and hospitals (both public and private) in Navarre were the primary source of data, while secondary sources were: independent general practitioners, health centre paediatricians and the Child-Youth Diabetics Parents' Association of Navarre. The degree of ascertainment was 97.8%. Average annual incidence of diabetes detected was 9.54/100000 (95% CI 8.2-11.1) in the 0-14 year-old group. The least incidence was observed in 1976 and highest in 1990. The incidence in males (9.71/100000) was higher than in females (7.83/100000). The highest incidence was observed in the 10-14 year-old group (13.70/100000) when analysed by groups. No seasonal variation in the onset of diabetes was observed. These results suggest a significant increase in the incidence of type 1 diabetes between 1975 and 1991. PMID- 9202799 TI - Factors predicting cerebral edema in young children with diabetic ketoacidosis and new onset type I diabetes. AB - We have attempted to identify any characteristics which could be used to predict the development of cerebral edema in four children under 5 years of age with new onset insulin-dependent diabetes mellitus and diabetic ketoacidosis. We retrospectively analysed and compared the concentration of serum sodium (corrected for serum glucose value) and effective serum osmolality of these 4 children with values of 10 age-matched controls with new onset insulin-dependent diabetes mellitus who did not develop cerebral edema during treatment of diabetic ketoacidosis. The initial serum sodium values of the two groups were not statistically different. Patients who developed cerebral edema had lower initial serum glucose values and effective serum osmolality. During treatment, patients who developed cerebral edema had consistently lower mean serum sodium and osmolality than controls at each 4-h interval after the first 4 h of therapy. Serum sodium and osmolality declined progressively after the initiation of therapy in cerebral edema patients, while remaining stable in controls. These data suggest that children who develop cerebral edema during treatment for diabetic ketoacidosis initially may have a relatively normal serum osmolality and subsequently develop progressive hyponatremia and/or a trend of declining serum sodium before developing cerebral edema. PMID- 9202801 TI - Plasma and tissue levels of lipids, fatty acids and plasma carnitine in neonates receiving a new fat emulsion. AB - This study was undertaken to compare Intralipid with a new fat emulsion containing gamma-linolenic acid and carnitine, named Pediatric Fat Emulsion 4501, in neonates with regard to lipid and carnitine metabolism over a short period of total parenteral nutrition. There were 10 neonates in each group and they tolerated the total parenteral nutrition well. In spite of the gamma-linolenic acid supplementation in the new emulsion, arachidonic acid decreased significantly in plasma lipid esters and adipose tissue in both groups after 5 d of treatment. Also, there was a decrease in plasma docosahexaenoic acid which was more pronounced in the treatment group. The relative percentage values of linoleic and linolenic acids in adipose tissue were increased, indicating that newborns have a rapid accretion of fatty acids. Plasma-triglycerides were effectively cleared during the periods without fat infusion. In the group that received Pediatric Fat Emulsion 4501 the means of both free and total plasma carnitine concentrations increased significantly, whereas they tended to decrease in the Intralipid group. PMID- 9202802 TI - Thermal control of the newborn: knowledge and practice of health professional in seven countries. AB - Hypothermia is a common problem in neonates, particularly in developing countries where it is an important contributory factor to neonatal mortality and morbidity. An evaluation of the knowledge and practices of health professionals on the thermal control of newborns was carried out in seven countries: Brazil, India, Indonesia, Kazakhstan, Mozambique, Nepal and Zimbabwe. The evaluation, conceived as a preliminary phase for a one-day training course on thermal control, involved 28 health facilities and 260 health professionals (61 doctors and 199 nurses and midwives). It included an assessment of thermal control practices carried out in each health facility by external investigators and a questionnaire on knowledge about thermoregulation administered to health professionals involved in newborn care. The findings of the evaluation were consistent across countries and showed that thermal control practices were frequently inadequate in the following areas: ensuring a warm environment at the time of delivery; initiation of breastfeeding and contact with mother, bathing; checking the baby's temperature; thermal protection of low birth weight babies, and care during transport. Knowledge on thermal control was also insufficient, especially concerning the physiology of thermoregulation and criteria for defining hypothermia. During the one-day course that followed the evaluation, participants were able to recognize the existing gaps and to identify appropriate interventions. Knowledge and practice on the thermal control of the newborn are currently insufficient. However, awareness of the importance of thermal control and basic knowledge on thermal regulation and thermal protection can be easily acquired and on this basis motivation for improving thermal control practices can be developed. PMID- 9202803 TI - Neonatal hearing screening with an automated auditory brainstem response screener in the infant's home. AB - Universal neonatal hearing screening is essential if all infants with congenital or perinatally acquired hearing impairment are to begin treatment before the age of 6 months to facilitate development of speech, language, communication and academic skills. Screening cannot always take place in hospital because of the increase in very short-stay deliveries. Therefore screening in the home may be necessary to achieve a high level of screening. We describe a feasibility study with an automated auditory brainstem response (AABR) screener in the infant's home as part of the service offered by the Well Baby Clinics in the Netherlands. Of the 277 infants who completed the screening 266 had the results "pass", 7 "refer" and 4 had inconclusive results. The mean time needed per screening was 18 min. This study shows that neonatal hearing screening by nurses using an AABR infant screener in the home is feasible. PMID- 9202804 TI - The effect of aerosolized furosemide in infants with chronic lung disease. AB - To investigate whether aerosolized furosemide would improve pulmonary function in premature infants with chronic lung disease, we enrolled eight preterm ventilator dependent infants in a cross-over, double-blind, placebo-controlled trial. Either aerosolized furosemide (2 mg/kg) or placebo (0.9% saline) was administered, and serial pulmonary function tests were performed before and at 1 and 2 h after each inhalation. After furosemide inhalation, static respiratory compliance increased significantly by 24.3% and 23.2% as percentage change from the baseline value at 1 and 2 h (p = 0.014 and 0.022, respectively). Also, tidal volume increased significantly by 33.8% and 28.7% at 1 and 2 h, respectively (p = 0.004 and 0.009). In contrast, no changes were observed in them after placebo inhalation. Total respiratory resistance was unchanged after both furosemide and placebo inhalation. There were no differences in urine volume in two groups. These data suggested that aerosolized furosemide improved pulmonary function in infants with chronic lung disease without excessive diuresis. PMID- 9202805 TI - A case of Kawasaki disease with coronary artery aneurysms documenting Yersinia pseudotuberculosis infection. AB - A case of a 5-year-old boy who fulfilled all the criteria for Kawasaki disease (KD) was described. He had associated bilateral coronary artery aneurysms. Our study revealed the isolation of Yersinia pseudotuberculosis in stool cultures, and the elevation and seroconversion of the agglutination antibody titres, and hence he was diagnosed as Y. pseudotuberculosis infection-positive. We also demonstrated the positive mitogenic activity of the culture supernatant of the isolated bacterium from the patient and detected Y. pseudotuberculosis-derived mitogen by PCR. This case therefore suggests that Y. pseudotuberculosis might be closely related to the cause of KD. PMID- 9202806 TI - Cerebral infarction in incontinentia pigmenti: the first report of a case evaluated by single photon emission computed tomography. AB - A 2-month-old girl with incontinentia pigmenti presented with acute-onset right handed focalized seizures and subsequent seizure generalization. Computed tomography, magnetic resonance imaging and single photon emission computed tomography results indicated that she had multiple cerebral infarctions. These findings suggest that incontinentia pigmenti should be included among the neurocutaneous syndromes associated with ischemic strokes in childhood. This is the first report of a case with incontinentia pigmenti associated with cerebral infarction evaluated by single photon emission computed tomography. PMID- 9202807 TI - Germ cell tumours in a brother and sister. AB - In familial germ cell tumour cases, a normal chromosomal karyotype pattern is rare. We report the findings of germ cell tumours in two siblings with a normal chromosomal karyotype. One of these patients had dysgerminoma in the right ovary and was treated successfully for this. At present, she is 23 years old and has two daughters. The other patient is a 15-year-old boy, who is the brother of the first patient and has mediastinal embryonal carcinoma. Although ultrasonography of the testes showed irregularity in the shape and non-homogeneity of the parenchyma, histopathological examination was found to be normal at the time of diagnosis. At present, he is doing well and his chemotherapy is continuing. Both of them have a normal chromosomal karyotype, 46, XX and 46, XY, respectively. We suggest that children who have a sibling with germ cell tumour should be carefully assessed for development of another germ cell tumour. PMID- 9202808 TI - Nutrition in disabled children. PMID- 9202809 TI - Autoimmunity and congenital permanent diabetes mellitus. PMID- 9202810 TI - Recurrent abdominal pain and Helicobacter pylori: a comment. PMID- 9202811 TI - Fetal alcohol syndrome. PMID- 9202812 TI - Effect of preseasonal enzyme potentiated desensitisation (EPD) on plasma-IL-6 and IL-10 of grass pollen-sensitive asthmatic children. AB - EPD is a method of preventive immunotherapy which employs b-glucuronidase as a biological response modifier. Plasma IL-6 and IL-10 were measured before a single injection of EPD, 24 hours later and 15 days after in a group of 17 children suffering from grass pollen asthma. 17 normal untreated children were used as controls. Although the study was conducted before the grass pollen season when the allergic children were free of symptoms, their plasma IL-6 and IL-10 were significantly elevated before the injection of EPD. 24 hours after treatment the plasma IL-10 had increased significantly and there was also a slight rise in IL 6. 15 days after treatment IL-6 had fallen to normal but IL-10 was still elevated. These findings suggest antigen-specific and non-specific mechanisms by which EPD may produce clinical improvement. PMID- 9202813 TI - Epidemiology: a critical tool for infection control professionals. PMID- 9202814 TI - Nosocomial infections in pediatric patients with burns. AB - BACKGROUND: Nosocomial infections (NI) are believed to occur more commonly in patients with burns than in patients undergoing surgery, but benchmark rates have not been well described, and widely accepted definitions of NI in patients with burns are not available. We present a clinically useful set of definitions for NI for the pediatric burn population and provide benchmark infection rates for NI at selected sites. METHODS: Centers for Disease Control and Prevention definitions were modified to more accurately describe nosocomial burn infection and secondary bloodstream infections (BSI) in the burn population. A surveillance system was developed and included calculation of NI rates by 1000 patient or device days, stratified into one of three risk groups (< 30% burn injury, 30% to 60% burn injury, and > 60% burn injury). All patients with acute burns admitted from January 1990 to December 1991 were included, and NI rates were calculated for burn infection, primary and secondary BSI, ventilator-related pneumonia and urinary catheter-related urinary tract infection (UTI). RESULTS: Overall 12.5% of patients with central venous catheters had development of primary BSI for a rate of 4.9/1000 central venous catheter-days. Incidence of secondary BSI was 5.8% of patients for a rate of 5.3/1000 patient-days. Incidence of burn infection was 10.1% of patients for a rate of 5.6/1000 patient-days. Incidence of ventilator related pneumonia was 17.5% of patients for a rate of 11.4/1000 ventilator-days. Incidence of urinary catheter-related UTI was 17.9% of patients, for a rate of 13.2/1000 urinary catheter-days. When rates were stratified by risk groups, incidence increased with increasing burn size for secondary BSI (p < or = 0.0001) and urinary catheter-related UTI (p = 0.08), although rates based on number of patient-days or device-days more accurately reflected risk of infection over time. CONCLUSIONS: Infection remains a cause of significant morbidity and death for patients with burns. The definitions and benchmark rates reported here may be useful in evaluation of NI surveillance strategies and calculation of infection rates, which could then be used to evaluate current treatment modalities and improve outcomes for the burn population. PMID- 9202815 TI - Detecting pediatric nosocomial infections: how do infection control and quality assurance personnel compare? AB - OBJECTIVE: To compare how well infection control (IC) and quality assurance (QA) personnel in a specialty setting identify the presence, type (nosocomial or community-acquired), and (if nosocomial) site of infection. METHODS: In 1994, we mailed a survey that included 21 pediatric case histories to IC and QA personnel in pediatric settings in the United States (children's hospitals and medical school-affiliated hospitals with pediatric wards of > 30 beds). From the case histories presented, the respondents were asked to determine whether an infection was present and, if so, whether it was nosocomial or community-acquired. If the infection was nosocomial, the respondent was asked to determine the site of the infection (e.g., urinary tract, bloodstream). RESULTS: From the 289 hospitals to which surveys were mailed, 131 respondents (45.3%) completed 212 surveys. Of the 212 returned surveys, 120 (56.6%) were completed by IC personnel and 92 (43.4%) were completed by QA personnel. Among the 183 respondents from acute care pediatric settings, 92.3% of IC personnel (96/104) and 54.4% of QA personnel (43/79) correctly identified at least 75% of the nosocomial infections (n = 14; p < 0.0001). IC and QA personnel were similar in ability to identify community acquired infection (88/104 vs 70/79, respectively; p = 0.436). IC personnel were significantly more likely than QA personnel to accurately identify the following sites of infection: respiratory tract infection without secondary bloodstream infection, necrotizing enterocolitis, urinary tract infection with and without secondary bloodstream infection, primary bloodstream infection, surgical site infection, gastroenteritis, esophagitis, and clinical sepsis. CONCLUSIONS: Overall, IC personnel were more accurate than QA personnel in determining whether a nosocomial infection was present and in correctly determining most sites of infection. Both IC and QA personnel had difficulty identifying venous infection and respiratory tract infection with secondary bloodstream infection. Both IC and QA personnel could thus benefit from more concise definitions or further training in detection of these sites of nosocomial infections. In addition, QA personnel did not perform overall as well as IC personnel in identifying nosocomial infections and their sites; this finding suggests the need for QA personnel to be provided specific training on detection of nosocomial infections and validation of their ability to do so. Nosocomial infection surveillance should be the responsibility of those trained and proved capable of detecting these infections. PMID- 9202816 TI - Blood contacts in the operating room after hospital-specific data analysis and action. AB - BACKGROUND: There has been much work recently to quantify risk of blood exposures among operating room personnel. Little has been done to show the outcome of preventive strategies. Three hospitals implemented a variety of changes after detailed feedback on blood contact data. This report follows those hospitals to document changes in blood contact rates. METHODS: Each hospital reviewed detailed data on blood exposures and developed a range of strategies to reduce contacts. In a second data collection period, uniformly trained circulating nurses sought information during surgical procedures on blood contacts among staff. Data were collected on all blood contacts and surgeries during which they occurred. These data were then compared with data from the previous study period, before changes in practices. RESULTS: All blood contacts combined and in each hospital decreased significantly in the second data collection period. Percutaneous exposures also consistently decreased, but did not reach statistical significance. The distribution of types of contact changed, with percutaneous exposures representing a larger proportion of contacts seen in the second period. Similar anatomic locations, devices, and characteristics of surgeries were associated with blood contacts in both periods. DISCUSSION: Specific data provided to operating room personnel motivated the development of specific strategies, although the influence of feedback alone versus specific interventions can not be separated. Analysis and generation of hospital-specific data on blood exposures among operating personnel may have a positive influence in lowering the risk of blood exposures in this population group. PMID- 9202817 TI - Hepatitis B vaccine booster dose: low-dose recombinant hepatitis B vaccines as a booster dose. AB - BACKGROUND: The timing and best regimen for a booster dose of hepatitis B vaccine have not been determined. METHODS: Two studies were conducted to determine the response to a booster dose of 5 micrograms recombinant hepatitis B vaccine. In the first study, a 5 micrograms (0.5 ml) dose of Recombivax HB was administered intramuscularly 38 months after the initial dose to 71 volunteers. In a second study, we offered a 5 micrograms dose recombinant hepatitis B vaccine, either Recombivax HB (0.5 ml) or Engerix B (0.25 ml), to students who had previously been immunized with three doses of vaccine. RESULTS: In the first study, among the 44 persons for whom postbooster sera were available, the geometric mean concentration of anti-hepatitis B surface antigens increased from 42 to 2090 mIU/ml after the 5 micrograms (0.5 ml) dose of Recombivax. In the second study, after a 5 micrograms (0.5 ml) dose of Recombivax, the geometric mean concentration increased from 43 to 990 mIU/ml (n = 48), and in the group that received a 5 micrograms (0.25 ml) dose of Engerix B, the concentration increased from 83 to 2337 mIU/ml (n = 45) (p = 0.18 for postdose concentrations). CONCLUSION: A 5 micrograms dose of recombinant vaccine results in an excellent booster response at a cost one fourth to one half that of a full 1 ml dose of vaccine. PMID- 9202818 TI - Side effects associated with pneumococcal vaccination. AB - BACKGROUND: Pneumococcal disease is a major cause of morbidity and death, especially among elders and other people at high risk. In spite of long-standing national recommendations for its use, pneumococcal vaccine is underused, with 70% or more of targeted persons as yet unimmunized. Concern about side effects is a barrier to successful vaccine delivery. METHODS: Persons attending a walk-in pneumococcal vaccination clinic were surveyed by use of structured telephone interviews. They were asked about health characteristics and local and systemic symptoms experienced during the week after their vaccination (postvaccination period). These responses were compared with the symptoms they reported for the 7 days immediately preceding their interview (the comparison period). RESULTS: A total of 1006 persons were interviewed a mean of 65.4 days after their vaccination. They had an average age of 69.9 years, and approximately 95% were in a high-risk group targeted for pneumococcal vaccination. For all systemic symptoms including fever, rash, myalgias, fatigue, malaise, and headache, subjects reported similar or lower rates during the postvaccination week than during the comparison week. Local reactions occurred in 28.2% of subjects. These local symptoms were mild to moderate for more than 90% of subjects and rarely resulted in the need to decrease the use of their arm. CONCLUSION: Pneumococcal vaccination was not associated with an increase in systemic symptoms but was associated with mild to moderate local symptoms in about one fourth of vaccine recipients. These findings should help health care providers and their patients address an important barrier to pneumococcal immunization. PMID- 9202819 TI - Status of tuberculosis infection control programs at Texas hospitals, 1989 through 1991. AB - BACKGROUND: Paralleling the resurgence of tuberculosis (TB) in the United States, the reported number of persons with TB in Texas increased by 33% during 1985 through 1992, the third largest rise among all the states. This increase prompted us to survey hospitals in Texas to determine their degree of compliance with recommendations in the Centers for Disease Control and Prevention TB guidelines. METHODS: In April 1992, we mailed a voluntary questionnaire about TB infection control practices, health care worker tuberculin skin testing procedures, and Mycobacterium tuberculosis laboratory methods to a convenience sample of hospitals in Texas. RESULTS: Of 180 hospitals surveyed, 151 (83%) returned completed questionnaires. Of these, 90 (60%) were nonteaching community hospitals; 28 (19%) were teaching community hospitals; 13 (9%) were university affiliated hospitals; and 20 (13%) were other hospitals. The number of hospitals to which patients with TB were admitted increased from 98 (65%) in 1989 to 122 (81%) in 1991. Respondent hospitals had a mean of 183 acute care beds (median 100, range 5 to 999), 6 acid-fast bacillus isolation rooms (median 2, range 0 to 57) and 7.5 admissions/year of patients with TB (median 2, range 0 to 202). Of hospitals responding to specific questions, 20% (27/137) admitted patients with multidrug-resistant TB, 18% (25/140) reported not having any acid-fast bacillus isolation rooms, and 28% (35/125) had no rooms meeting all of the Centers for Disease Control and Prevention criteria for acid-fast bacillus isolation (negative air pressure, > or = 6 air changes per hour, and air directly vented to the outside). The tuberculin skin test conversions among health care workers rose from 246 (0.6%) in 1989 to 547 (0.9%) in 1991. CONCLUSION: Although the number of Texas hospitals admitting patients with TB increased during 1989 through 1991, many facilities still did not have infection control practices consistent with the 1992 Centers for Disease Control and Prevention TB guidelines. PMID- 9202820 TI - A retrospective on infection control. Part 1: Nineteenth century--consumed by fire. PMID- 9202821 TI - Acquisition of hepatitis C by a conjunctival splash. PMID- 9202822 TI - Health hazards in nursing and health care: an overview. PMID- 9202823 TI - Applied epidemiology and environmental health: emerging controversies. AB - This review article assesses the state of the science in environmental epidemiology, not by summarizing current scientific findings but rather by examining conceptual controversies in the study of how environmental factors influence human health. This approach seems necessary because environmental epidemiology presently stands at a crossroads-in fact, at a number of overlapping crossroads. The field teems with epistemologic debates concerning appropriate paradigms for framing research questions, interpreting data, and applying research findings to policy. The present review focuses on emerging controversies related to three questions: What is considered "environmental"? What counts as credible research in environmental epidemiology? And what does "applied epidemiology" mean in the context of environmental health? The goal is to organize the presently fragmented critical literature on these issues and to promote productive dialogue by identifying central themes in current conceptual debates. PMID- 9202824 TI - Obstacles encountered in application of the Centers for Disease Control and Prevention guidelines for control of tuberculosis in a large dental center. AB - BACKGROUND: The Centers for Disease Control and Prevention (CDC) and the Occupational Safety and Health Administration have designated five categories of workplaces as carrying higher than normal risk for exposure to tuberculosis (TB); "health care facilities" is one of these categories. To assist all health care facilities in developing an appropriate and effective control plan, the CDC has listed various components to be included in the overall plan-however, the components needed cannot be determined until the level of risk has been determined. The published criteria for risk assessment are more appropriately applicable to a hospital-based facility. In complying with CDC's guidelines and adopting the recommended components of the TB control program at a large, educational, ambulatory care dental facility, several obstacles were identified. METHODS: As part of the risk assessment for TB transmission and to determine the significance of purified protein derivative of tuberculin (PPD) skin-test conversions observed among the student and employee populations, we implemented a strategy that consisted of surveying all accredited dental schools in the country, performing a controlled PPD screening study involving predoctoral dental students in their junior year, and reviewing and evaluating all patient registration records for the same period. RESULTS: Forty-four percent of the dental schools contacted agreed to participate in the survey. Of these, 58% had no information available on student PPD conversion rates and 29% either had no data available yet or were unwilling to share their information. Three schools (12%) that had some data and were willing to share it reported PPD conversion rates for faculty, students, or staff of 1% to 2%. The student PPD study showed a 10.7% conversion rate, but the registration record reviews showed no convincing evidence of patients with active TB having been registered at the dental school for the period of the student PPD study. CONCLUSION: Development of a TB control program relies heavily on assessment of risk within a health care facility. The sporadic reports of PPD conversion rates among dental care workers are not adequate to determine the magnitude of exposure to TB in educational dental settings. Further studies are necessary to establish the true risk and to assist dental care facilities in developing TB control programs. PMID- 9202825 TI - Benefit of two-step PPD testing of new employees at a New York City hospital. AB - BACKGROUND: Recent concern about nosocomial transmission of tuberculosis has led hospitals to scrutinize employee tuberculin conversion rates. The Centers for Disease Control and Prevention recommends two-step testing of new employees to limit the booster phenomenon. The cost of such a program and its subsequent yield have not recently been examined. METHODS: Employee health records were retrospectively reviewed of persons hired from 1993 and 1994 at St. Clare's Hospital in New York City, all of whom received two-step testing at time of initial employment. RESULTS: Of 262 new employees, 107 (41%) had positive tuberculin results on initial testing. The results of 15 (9.7%) of the remaining 155 patients became positive on two-step testing administered 1 week later. Persons with a positive second test result were significantly more likely to be male or foreign born or to have received previous bacille Calmette-Guerin vaccination. Identification of these 15 persons and exclusion of them from probable subsequent conversion prevented an almost 50% increase in the annual conversion rate at our hospital, from 3.2% to 4.7%. CONCLUSION: Two-step tuberculin testing is an essential means of identifying persons with a baseline positive tuberculin test result, thus allowing accurate reporting of subsequent employee tuberculin conversions. PMID- 9202826 TI - HICPAC guidelines for isolation precautions: Hospital Infection Control Practices Advisory Committee. PMID- 9202827 TI - Use of a triple-stain technique to detect viability and acrosome reaction in deer spermatozoa. AB - A procedure is described for simultaneously evaluating of sperm viability and acrosomal status in fixed deer spermatozoa. This technique, which is a modification of the triple-stain technique (TST) developed for human sperm, utilizes trypan blue to identify the percentage of live sperm in culture and subsequent staining of fixed sperm to differentiate the acrosome. Four classes of deer sperm can be distinguished with the TST: (1) live unreacted sperm, (2) live acrosome-reacted sperm (true acrosome reaction), (3) dead unreacted sperm, and (4) dead acrosome-reacted sperm (false acrosome reaction). The study shows that the acrosome status can be detected either by TST stain or by phase-contrast microscopy (r = .971, p < .001) and that TST does not differ from 0.4% trypan blue in its ability to identify live and dead sperm (r = .995, p < .001). This staining procedure enables differentiation of spermatozoa that have undergone a true acrosome reaction (TAR) from those that have undergone a false acrosome reaction (FAR). In this way, incubation of deer spermatozoa for 30 min at 37 degrees C in the presence of calcium ionophore A23187 increased the proportion of spermatozoa undergoing a TAR. Sperm incubated in the absence of A23187 undergo the TAR at a much lower rate (p < .001) than those incubated in the presence of ionophore. This stain procedure was also used to study the time course of the true acrosome reaction of deer sperm in vitro. Incubation of deer spermatozoa for up 24 h at 37 degrees C resulted in a decrease in the percentage of live acrosome intact spermatozoa and a simultaneous increase in the percentage of spermatozoa categorized as having undergone a TAR and FAR. PMID- 9202828 TI - Generation of reactive oxygen species by leukocytes and sperm following exposure to urogenital tract infection. AB - The association between male urogenital tract infection (UTI) and infertility is still controversial. Oxidative stress caused by reactive oxygen species (ROS) in semen has been suggested to be an important factor in the etiology of poor sperm function through peroxidative damage to the cell membrane. This study explored the potential association between the ROS generation and UTIs by examining ROS production by sperm and seminal leukocytes in response to various infectious and cytokine stimulating factors. Semen and blood samples were obtained from 17 normal donors. Highly motile pure sperm, poorly motile sperm, and polymorphonuclear leukocytes (PMN) were isolated and exposed to various infectious and stimulating factors, including lipopolysaccharide (LPS), gamma interferon (gamma-IFN), tumor necrosis factor-alpha (TNF-alpha), granulocyte, macrophage-colony stimulating factor (GM-CSF), and phorbol myristate acetate (PMA). ROS generation was determined by measuring luminescence in a luminometer. In this study, purified PMNs produced high levels of ROS, and this production was markedly enhanced in the presence of cytokines, LPS, as well as PMA. Pure motile sperm produced low levels of ROS, and ROS production was not enhanced by addition of bacterial products or cytokines. In conclusion, PMNs in semen are the major source of ROS, and ROS production by these cells is enhanced by bacterial products and cytokines. Detection of activation markers and/or soluble factors produced by activated leukocytes in the urogenital tract or semen could enhance the diagnosis and lead to improved therapy of male infertility due to subclinical genital tract infections. PMID- 9202829 TI - Unusual features of the sperm head differentiation in Mabuya quinquetaeniata. AB - Sperm head differentiation in Mabuya quinquetaeniata agrees in the main features with that of Agama adramitana, Agama blandfordi, and Acanthodactylus boskianus. The development of subacrosomal lateral canals, the disappearance of translucent medulla, and the existence of unilateral dense acrosome are new findings of the present investigation. PMID- 9202830 TI - Clinical application of a new stain to detect acrosome-reacted sperm for predicting polyspermic fertilization in IVF-ET. AB - The aim of this study was to evaluate the efficacy of Blutstan staining of human spermatozoa for predicting the spermatozoa capacity of fertilization in human IVF. Blutstan, a prestained glass slide coated with dyes, is able to identify activated sperm quickly and easily. Acrosomal reactivity of spermatozoa was evaluated with this slide glass at the insemination of IVF of 30 couples. There was significant correlation between the Blutstan reactivity and the fertilization rate (r = .52, p < .01). Furthermore, spermatozoa with high Blutstan reactivity were fertilized oocytes polyspermy. This method was rapid, simple, and useful for detecting activated sperm and predicting for the polyspermic fertilization in clinical setting. PMID- 9202831 TI - Effect of centrifuge speed, refrigeration medium, and sperm washing medium on cryopreserved sperm quality after thawing. AB - Cryopreservation and subsequent thawing of semen for assisted reproductive procedures decreases sperm motility; motility further reduces when the cryoprotectant medium is removed because of the osmotic shock and centrifugation done to prepare the sperm. To compare motility and thus pregnancy rates, this study examined the effects of adding an additional refrigeration medium and three different centrifugation speeds for sperm preparation. Semen samples from 16 healthy normal volunteers were obtained by masturbation after 48 h of abstinence. Sperm motility and other motion characteristics were analyzed with a computer assisted semen analyzer before freezing, after thawing, and after centrifugation. Each sample was divided into 6 aliquots and frozen using the liquid nitrogen vapor method. After thawing, human tubal fluid (HTF) with 5% human serum albumin was added to 3 aliquots, and refrigeration medium (identical to freezing medium without glycerol) was added to the remaining 3 tubes for each specimen. The tubes containing the two media were then washed by centrifugation at 100 g, 300 g, and 500 g for 10 min. Aliquots with refrigeration medium did not significantly differ from those with HTF for percent motility, curvilinear velocity, straight-line velocity, and amplitude of lateral head displacement at any centrifugation speed. Motile sperm count was significantly greater only at 100 g and 300 g for refrigeration medium (p = .02 and .01) and HTF (p = 0.001); at 300 g, average path velocity in refrigeration medium aliquots (p = .01) and linearity in HTF (p = .01) were greater. The results indicated that the reduction in motility and other motion characteristics probably cannot be overcome by changing factors such as the sperm preparation medium or centrifugation speed. More effective cryopreservation techniques or preparation methods that eliminate centrifugation need to be developed. PMID- 9202832 TI - Prostasome fraction of human seminal plasma prevents sperm from becoming acrosomally responsive to the agonist progesterone. AB - Seminal plasma prevents human sperm from becoming acrosomally responsive. These experiments tested the idea that the inhibitory activity of seminal plasma is contained in the particulate prostasome fraction. Most of the inhibitory activity was sedimentable (105,000 g, 2 h) and the majority of the recovered activity was in the prostasome fraction. The recovery of inhibitory activity in the prostasome fraction (42% of the activity in unfractionated seminal plasma) was similar to the recovery of cholesterol in that fraction (41%), consistent with cholesterol's role as the major inhibitor in seminal plasma. To test whether components of the prostasome fraction bind to sperm, the prostasome fraction was made fluorescent with fluorescein isothiocyanate or with the acetoxymethyl ester of 2',7'-bis-(2 carboxyethyl)-5(6)-carboxyfluorescein. No labeled material was seen to bind to sperm, suggesting that exchange of cholesterol between prostasomes and sperm takes place through the aqueous phase or at the time of vesicle-sperm collision. PMID- 9202833 TI - IgM and IgG in the human male genital tract. AB - In human seminal plasma it is possible to measure the levels of IgG and IgA, while IgM has been detected in trace amounts in only a few subjects. The source of these antibodies is unknown, although, under certain conditions, they seem to be produced in the sex tissues. Male human genital tract was processed for the immunohistochemical demonstration of IgG and IgM to verify the presence and the source of these antibodies. Only the epithelia of the prostate gland and urethra showed positivity to the immunohistochemical reaction, indicating the presence of IgM-secreting epithelial cells. Clusters of IgM- and IgG-positive immunocompetent cells were also observed in the subepithelial layers of all organs studied. The findings suggest that IgM are present in mucosal surfaces as a result of active secretion, as well as secretory IgA, whereas most of the IgG present in the human seminal plasma are probably derived by transudation process. PMID- 9202834 TI - Characteristics of men and women with circulating antisperm antibodies in a combined infertility clinic in Kuwait. AB - Antisperm antibodies were determined in the sera of 250 infertile couples and 100 puerperal women as controls using the immunofluorescence technique. Couples with significant circulating antisperm antibodies were placed on low-dose prednisolone 5 mg daily for 3-6 months. Initial routine semen analysis and hypoosmotic swelling test were done and repeated after 3 months of therapy. The incidence of antisperm antibodies (ASA) was 18.8 and 17.6% in the men and women, respectively, compared to 4% in the women controls (p < .02). In the men, the main determinants (with incidence) of ASA included smoking (33.9%), past history of sexually transmitted disease (33.3%), surgery to genital tract (28.6%), trauma (27.3%), and unexplained infertility (18.5%). In women whose husbands had antisperm antibodies the incidence of circulating antisperm antibodies was 38.3%, while endometriosis and thyroid dysfunction had incidence of antisperm antibodies of 21.4 and 16.7%, respectively. In the 27 (10.8%) case of unexplained infertility, the incidence of antisperm antibodies was 22.2%. High follicle-stimulating hormone (FSH) in the men and low midluteal-phase progesterone in the women were associated with increased expression of antisperm antibodies. Antisperm antibodies adversely affected quality of sperm. Low-dose prednisolone significantly reduced the titer of antisperm antibodies and improved the sperm parameters and conception rate. PMID- 9202835 TI - Effects of pentoxifylline on sperm motility in normogonadotropic asthenozoospermic men. AB - Forty-seven normogonadotropic men with idiopatic asthenozoospermic were divided at random: group I (N = 22) received placebo and group II (N = 25) received 1200 mg of pentoxifylline/day during 6 months. Semen analysis was performed basal and at 3 and 6 months of the study period. No statistical changes in serum hormone concentration were found, nor in volume, sperm counts, viability, and morphology before and after treatment. Sperm motility increased following pentoxifylline treatment after 3 and 6 months from 25.5 (21.0-30.0) to 35.5 (31.5-39.0) (p < .00001) and to 42.0 (38.0-46.0) (p < .00001), respectively. Although in the placebo control cases some changes were observed in the sperm motility, they were less significant. Furthermore, progressive motility only in grade A increased with pentoxifylline from 2.5 (0.0-6.0) to 12.0 (6.0-19.5) (p < .001) at 3 months and to 22.5 (17.0-26.0) at 6 months (p < .00001). In conclusion, pentoxifylline had an additional effect rather than placebo and was useful treatment in these cases of male factor infertility. PMID- 9202836 TI - Pelvic floor muscles and sphincters during erection and ejaculation. AB - The pelvic floor muscles (puborectalis [PR], levator ani [LA]) and external anal (EAS) and urethral sphincters (EUS) were studied during erection and ejaculation in 12 mongrel dogs. The animals were anesthetized and the response of PR, LA, EAS, and EUS to electroejaculation (EE) was evaluated. Penile tumescence and rigidity as well as increase of the EMG activity of the aforementioned muscles and sphincters occurred. The rigidity and EMG activity increased as the voltage and current of EE continued to increase until ejaculation occurred at a mean voltage of 11.1 +/- 1.2 V and current of 122.4 +/- 14.4 mA. The increased PR activity might express the prostatic secretions into the posterior urethra. LA contraction seems to elevate the prostate and partially straightens the prostato membraneous urethral kink that might occur during erection. The EAS and EUS contractions are believed to abort the urge to defecate or urinate and prevent leak of feces, flatus, or urine during coitus. The rhythmic EUS contraction at ejaculation might act as a "suction ejection pump," sucking the genital fluid into the posterior urethra while being relaxed and ejecting it into the bulbous urethra upon contraction. PMID- 9202837 TI - Amine neurotransmitter levels in male and female rats through developmental periods. AB - Monoamines (MA) such as dopamine (DA), norepinephrine (NE), and 5 hydroxytryptamine (5-HT) are now generally regarded as widely distributed and essential endogenous mediators contributing to the integration of reproductive physiology. MA measured in the hypothalamus tissue of male and female rats aged 1 to 90 days showed its own characteristic development pattern. Significant differences were observed at 5, 15, and 90 days of age in NE mean levels and at all ages except for 3 days of age in 5-HT mean levels. In contrast, no sex differences were seen in DA mean levels. PMID- 9202838 TI - Ca transport during contraction and relaxation in mammalian ventricular muscle. PMID- 9202839 TI - Sites of regulatory interaction between calcium ATPases and phospholamban. AB - In an effort to define the amino acids that are involved in functional interactions between phospholamban (PLN) and the Ca2+ ATPase of cardiac sarcoplasmic reticulum (SERCA2), we have co-expressed wild type and mutant forms of phospholamban with wild type and mutant forms of SERCA2, isolated microsomal fractions and measured Ca2+ dependence of Ca2+ transport. We have found that both charged and hydrophobic residues in the cytoplasmic domains of both PLN and SERCA2 make up the cytoplasmic interaction site. In SERCA2, this site is the linear sequence Lys-Asp-Asp-Lys-Pro-Val402: In PLN, the site is more diffuse and complex. Function was retained if the net charge over the first 20 amino acids was +1 or +2, but function was lost if the net charge was -3, -2, 0 or +3. Function was also lost if the long alkyl side chains of Val4, Leu7 or Ile12 were replaced with the methyl group of Ala. We have also obtained evidence that a site of functional interaction is present in the transmembrane domains of PLN and SERCA2. PMID- 9202840 TI - The relative phospholamban and SERCA2 ratio: a critical determinant of myocardial contractility. AB - Phospholamban is a regulatory phosphoprotein which modulates the active transport of Ca2+ by the cardiac sarcoplasmic reticular Ca(2+)-ATPase enzyme (SERCA2) into the lumen of the sarcoplasmic reticulum. Phospholamban, which is a reversible inhibitor of SERCA2, represses the enzyme's activity, and this inhibition is relieved upon phosphorylation of phospholamban in response to beta-adrenergic stimulation. In this way, phospholamban is an important regulator of SERCA2 mediated myocardial relaxation during diastole. This report centers on the hypothesis that the relative levels of phospholamban: SERCA2 in cardiac muscle plays an important role in the muscle's overall contractility status. This hypothesis was tested by comparing the contractile parameters of: a) murine atrial and ventricular muscles, which differentially express phospholamban, and b) murine wild-type and phospholamban knock-out hearts. These comparisons revealed that atrial muscles, which have a 4.2-fold lower phospholamban: SERCA2 ratio than ventricular muscles, exhibited rates of force development and relaxation of tension, which were three-fold faster that these parameters for ventricular muscles. Similar comparisons were made via analyses of left ventricular pressure development recorded for isolated, work-performing hearts from wild-type and phospholamban knock-out mice. In these studies, hearts from phospholamban knock-out mice, which were devoid of phospholamban, exhibited enhanced parameters of left-ventricular contractility in comparison to wild-type hearts. These results suggest that the relative phospholamban: SERCA2 ratio is critical in the regulation of myocardial contractility and alterations in this ratio may contribute to the functional deterioration observed during heart failure. PMID- 9202841 TI - Phosphorylation and regulation of the Ca(2+)-pumping ATPase in cardiac sarcoplasmic reticulum by calcium/calmodulin-dependent protein kinase. AB - In cardiac muscle, a membrane-associated Ca2+/calmodulin-dependent protein kinase (CaM kinase) phosphorylates the Ca(2+)-pumping ATPase in addition to its previously characterized substrates, phospholamban and Ca(2+)-release channel (ryanodine receptor). The phosphorylated amino acid in the Ca(2+)-ATPase has been identified as serine. Posphorylation of the Ca(2+)-ATPase is rapid and is reversible by a membrane-associated protein phosphatase, Ca(2+)-ATPase purified from cardiac SR underwent phosphorylation by exogenous CaM kinase, and the phosphorylated enzyme displayed twofold greater catalytic activity without alteration in its Ca(2+)-sensitivity. The phosphorylation of the Ca(2+)-ATPase was found to be isoform-specific in that the cardiac and slow-twitch skeletal muscle isoform (SERCA 2), but not the fast-twitch skeletal muscle isoform (SERCA 1), underwent phosphorylation by CaM kinase. Studies using SERCA 1 and SERCA 2 isoforms and their mutants expressed in a heterelogous cell system have resulted in i) confirmation of the isoform specificity of Ca(2+)-ATPase phosphorylation by CaM kinase, ii) identification of Ser38 as the site in SERCA 2 phosphorylated by CaM kinase, and iii) demonstration of phosphorylation-induced increase in Vmax of Ca2+ transport by the SERCA 2 enzyme. These observations suggest that in cardiac and slow-twitch skeletal muscle direct phosphorylation of the SR Ca(2+)-ATPase by the membrane-bound CaM kinase may serve to stimulate Ca2+ sequestration and therefore, the speed of muscle relaxation. PMID- 9202842 TI - Site-specific phosphorylation of a phospholamban peptide by cyclic nucleotide- and Ca2+/calmodulin-dependent protein kinases of cardiac sarcoplasmic reticulum. AB - Phospholamban (PLB), the regulator of the cardiac sarcoplasmic reticulum (SR) Ca2+ pump is specifically phosphorylated at Ser16 and Thr17 by cAMP-dependent protein kinase (PKA) and Ca2+/calmodulin-dependent protein kinase (CaMK), respectively. The regulation of this dual-site phosphorylation of amino acid residues in direct proximity is only poorly understood. In order to study the site-specific phosphorylation of PLB, we used a synthetic peptide (PLB-24) corresponding to the cytosolic part of the PLB monomer with the phosphorylation sites as a model substrate. PLB-24 possesses substrate properties as the native PLB as demonstrated by phosphorylation with exogenous, purified PKA, cGMP dependent protein kinase (PKG) and a type II CaMK (CaMKII). In isolated vesicles of cardiac SR there was a rapid phosphorylation of the peptide by the endogenous PKA (SR-PKA) and CaMK (SR-CaMK), but not under conditions that activate PKG. Both SR-PKA and SR-CaMK incorporated the same amount of 32P into PLB-24, 0.60 +/- 0.01 nmol 32P/mg SR protein and 0.61 +/- 0.03 nmol 32P/mg SR protein, respectively. Phosphorylation by SR-PKA was abolished by the specific PKA inhibitor (IC50 = 0.2 microM), whereas SR-CaMK phosphorylation was inhibited by calmidazolium (IC50 = 1.6 microM) and a CaMKII-specific inhibitor peptide (IC50 = 2.5 microM). Phosphorylation by SR-PKA was exclusively at Ser, whereas SR-CaMK phosphorylated only Thr. After simultaneous activation of both SR-kinases 32P incorporation into PLB-24 was additive and occurred at Ser as well as at Thr. Sequential activation of SR-PKA and SR-CaMK also caused the additive phosphorylation of PLB-24 independently of which kinase was activated first. Thus, at the monomeric level of PLB the respective phosphorylation site appears to be accessible to its related SR protein kinase in vitro even when the adjacent site is phosphorylated. PMID- 9202843 TI - Sodium-calcium exchange: recent advances. AB - Na-Ca exchange proteins are involved in Ca homeostasis in a wide variety of tissues. Unique Na-Ca exchangers have been identified by molecular biological approaches and it appears that these may represent a superfamily of ion transporters, similar to that identified for ion channels. Major advances in our understanding of these transporters have occurred in the past decade by combining molecular approaches with electrophysiological analyses. The regulatory and transport properties of Na-Ca exchangers are beginning to become understood in molecular detail. It also appears that the physiological roles of Na-Ca exchange may be quite complex. This brief review highlights some recent advances in Na-Ca exchange research obtained through the combination of molecular biological and electrophysiological approaches. PMID- 9202844 TI - Expression and function of the cardiac Na+/Ca2+ exchanger in postnatal development of the rat, in experimental-induced cardiac hypertrophy and in the failing human heart. AB - The diastolic and systolic dysfunction in the failing heart appear to be related to the altered Ca2+ handling of the cardiac myocyte. Disturbed Ca2+ handling might also affect influx and efflux of other ions, including Na+. In this context, the cardiac sarcolemmal Na+/Ca2+ exchanger represents an important exchange mechanism of Ca2+ versus Na+ transport across the sarcolemma. Expression and function of cardiac Na+/Ca2+ exchanger is highest in newborn rats and declines gradually in postnatal development. In pressure overload-induced hypertrophy, expression of cardiac Na+/Ca2+ exchanger is increased and translated into increased Na+/Ca2+ exchanger activity similar to the early phase of postnatal development in the rat. This suggests a common underlying mechanism in the control of Na+/ Ca2+ exchanger expression in the immature and the hypertrophied myocardium. Similar to experimental-induced hypertrophy, mRNA, protein and activity of Na+/ Ca2+ exchanger is increased in the failing human heart suggesting an increase in the number of functional exchanger molecules rather than an enhanced exchange rate by preexisting exchanger molecules. The potential functional implications of an increased cardiac Na+/Ca2+ exchanger activity in human heart failure may be limitation of diastolic intracellular Ca2+ overload. However, this may increase the arrhythmogenic potential of the failing heart, since additional Na+ influx via Na+/Ca2+ exchanger may affect the membrane potential. PMID- 9202845 TI - Plasma membrane calcium pump: structure, function and relationships. AB - The plasma membrane Ca-pump (134 kDa) is stimulated by calmodulin and by other treatments (exposure to acidic phospholipids, treatments with proteases, phosphorylation by protein kinases A or C, self-association to form oligomers). It is the product of four genes (in humans), but additional isoforms originate through alternative mRNA spicing. Most of the pump mass protrudes into the cytoplasm with three main units. The calmodulin binding domain is located in the C-terminal protruding unit. The domain is a positively charged segment of about 25 residues. The calcium-activated protease calpain activates the pump by removing its calmodulin binding domain and the portion C-terminal to it. The resulting 124 KDa fragment has been used to test the suggestion of an autoinhibitory function of the calmodulin binding domain. The latter interacts with two domains of the pump, one located close to the active site in the mid cytoplasmic protruding unit, the other in the first (N-terminal) protruding unit. The isoforms of the pump show variations in the regulatory domains, e.g., alternative mRNA splicing can eliminate the domain phosphorylated by protein kinase A, or alter the sensitivity of the pump to calmodulin. This occurs by inserting sequences rich in His between calmodulin binding subdomains A and B. The inserted domain(s) confer pH sensitivity to the binding of calmodulin. Calcium binding sites have been found in acidic regions preceding and following the calmodulin binding domain. PMID- 9202846 TI - Molecular mechanisms regulating the myofilament response to Ca2+: implications of mutations causal for familial hypertrophic cardiomyopathy. AB - In this chapter we consider a current perception of the molecular mechanisms controlling myofilament activation with emphasis on alterations that may occur in familial hypertrophic cardiomyopathy (FHC). FHC is a sarcomeric disease (100) with an autosomal dominant pattern of heritability (27, 51). There is a substantial body of evidence implicating missense mutations in the beta-MHC gene as causal for the development of this disease. Recently, mutations in genes of two thin filament regulatory proteins, cardiac troponin T(cTnT) and alpha tropomyosin (alpha-Tm), have also been linked to FHC. The commonality among the functional consequences of these mutations remains an important question. This review discusses how these pathological mutations may impact the activation process by disrupting critical structure function relations in both the thick and thin filaments. PMID- 9202847 TI - Ca(2+)-dependent and Ca(2+)-independent regulation of contractility in isolated human myocardium. AB - Changes in contractile force of the myocardium may depend on changes in the intracellular Ca2+ concentration, changes in the responsiveness of the myofibrils for Ca2+, or a combination of both. We investigated in isolated muscle strip preparations from human nonfailing and endstage failing hearts the influence of physical (changes in preload, stimulation rate, or rhythm), and pharmacological interventions (alpha- or beta-adrenoceptor-stimulation, endothelin) on developed force of contraction and the corresponding intracellular Ca2+ transients. METHODS: Isometric contraction, electrical stimulation, 37 degrees C. Simultaneous registration of force of contraction and intracellular Ca2+ transients (aequorin method). RESULTS: Increases in preload, alpha- and endothelin-receptor stimulation resulted in increases in force of contraction without increasing aequorin light emission. Increasing stimulation rate or increasing rest intervals resulted in parallel increases (nonfailing myocardium) or decreases (failing myocardium) of force of contraction and aequorin light emission. beta-Adrenoceptor-stimulation exerted inotropic and lusitropic effects in human failing myocardium associated with a large, overproportional increase in aequorin light emission. CONCLUSION: The human heart regulates intrinsic contractility via several subcellular mechanisms. Increases in preload (Frank Starling-mechanism) and alpha- or endothelin-receptor-stimulation enhance myocardial contractility by increasing the Ca2+ responsiveness of the myofilaments; rate- and rhythm-dependent modulation of the contractile state directly depend on changes in the intracellular Ca(2+)-transients; beta adrenoceptor stimulation results in an overproportional large increase in intracellular Ca2+ transients, probably due to additional cAMP-dependent Ca(2+) desensitizing effects on the level of the myofibrils. PMID- 9202848 TI - Calcium handling proteins in the failing human heart. AB - There is accumulating evidence that disturbed calcium homeostasis may play a key role in the pathophysiology of human heart failure. Because disturbed calcium handling could result from altered protein expression, levels of calcium handling proteins were quantitated by Western Blot analysis in failing and nonfailing human myocardium from hearts with endstage failing dilated or ischemic cardiomyopathy. Protein levels of the sarcoplasmic reticulum calcium release channel (ryanodine receptor) and of calcium storage proteins (calsequestrin and calreticulin) were similar in failing and nonfailing human myocardium. However, proteins involved in calcium removal from the cytosol were significantly altered in the failing human heart: 1) SR-Ca(2+)-ATPase, relevant for removal of calcium from the cytosol into the lumen of the sarcoplasmic reticulum, was decreased; 2) phospholamban, which inhibits the SR-Ca(2+)-ATPase in the basal unphosphorylated state, was slightly decreased; 3) the ratio of SR-Ca(2+)-ATPase to phospholamban was decreased; 4) the sarcolemmal Na(+)-Ca(2+)-exchanger, relevant for transsarcolemmal calcium extrusion was increased in the failing hearts. In summary, altered levels of proteins involved in calcium removal from the cytosol suggest an increase in transsarcolemmal calcium elimination relative to sarcoplasmic reticulum calcium removal. These findings support the concept that reduced function of the sarcoplasmic reticulum to accumulate calcium may reflect a major defect in excitation-contraction coupling in human heart failure. PMID- 9202849 TI - Role of cAMP in modulating relaxation kinetics and the force-frequency relation in mitral regurgitation heart failure. AB - The report is a discussion of previously published and newly analyzed results concerning the association between heart diseases and alterations in the force frequency relation (FFR). The optimum stimulation frequency of the FFR is measured and compared in isolated left ventricular myocardium from non-failing hearts with atrial septal defect, coronary artery disease (without and with insulin dependent diabetes mellitus) and from failing hearts with mitral regurgitation, or idiopathic dilated cardiomyopathy. Specifically, we examine the role of altered control of the excitation-contraction coupling system in blunting the force-frequency relation. We use the percent slope of the FFR as a measure of changes in the frequency sensitivity of this control. Our finding of a linear, direct relation between optimum stimulation frequency and % slope across all disease types suggests both parameters are coupled to the same underlying mechanism. To investigate the possible role of altered control of the calcium pump in this mechanism, we analyzed the detailed relation between isometric twitch relaxation kinetics and stimulation frequency in mitral regurgitation myocardium (MR). In the presence of 0.5 microM forskolin the depressed slope and optimum frequency of the FFR and the prolonged half-time of twitch relaxation were all restored to values found in non-failing myocardium. We use the kinetics of isometric twitch relaxation as an index of changes in pumping rate that occur in response to changes in stimulation frequency or in intracellular cyclic adenosine monophosphate concentration. A mathematical model based on the Hill relations for calcium pump uptake rate and for isometric tension as a function of intracellular pCa is developed to simulate isometric twitch relaxation in MR and non-failing myocardium. The success of this model in simulating non-failing and failing twitch relaxation supports a proposed mechanism for the prolonged relaxation time and depressed FFR in MR involving depressed protein kinase-A activity (due to lowered cAMP or to a defect in the Ser16 site of phospholamban) as a mechanism of altered control of the calcium pump in MR heart disease. PMID- 9202850 TI - Contributions of Ca(2+)-influx via the L-type Ca(2+)-current and Ca(2+)-release from the sarcoplasmic reticulum to [Ca2+]i-transients in human myocytes. AB - Experiments were performed to determine the relative contributions of direct Ca(2+)-entry through the L-type Ca(2+)-current and of Ca(2+)-release from the sarcoplasmic reticulum (s.r.) to the intracellular [Ca2+]i-transient in isolated human atrial and ventricular myocytes from patients with severe heart failure and from non-failing controls. Cells were isolated from explanted hearts of patients undergoing transplantation because of severe heart failure due to dilated or ischemic cardiomyopathy or from donor hearts which could not be transplanted for technical reasons. Ca(2+)-current densities were -2.1 +/- 0.6 pA/pF in atrial cells, -4.8 +/- 0.5 pA/pF in cells from patients with heart failure and -3.2 +/- 0.5 pA/pF in non-failing controls. [Ca2+]i-transients were significantly smaller in heart failure (370 +/- 33 nM) compared to ventricular cells from non-failing hearts (760 +/- 69 nM, p < 0.05). Atrial myocytes had average [Ca2+]i-transients of 505 +/- 38 nM. After incubation in ryanodine the average [Ca2+]i-transients were not significantly different between different cell types. The results indicate that the relative contribution of Ca(2+) released from the sarcoplasmic reticulum to the [Ca2+]i-transient is significantly smaller in heart failure. The absolute contribution of the L-type Ca(2+)-current to the transient seemed to be comparable in all cell types investigated. As the [Ca2+]i-transient in the presence of ryanodine was comparable in size in all cells, changes of the intracellular [Ca2+]i-transient in heart failure are mainly due to alterations of s.r. function in these cells. PMID- 9202851 TI - Molecular and cellular aspects of re-entrant arrhythmias. AB - In recent years it has become evident that myocardial tissue undergoes remodeling in diseased states such as myocardial infarction and hypertrophy which affects membrane channels, cell-to-cell coupling as well as the connective tissue matrix. Although the detailed mechanisms of ventricular arrhythmias in ventricular hypertrophy are not known, studies carried out by computer simulations or high resolution mapping of electrical activity have suggested a complex interaction between changing ionic currents at the level of the cell membranes, altered cell to-cell coupling and altered macroscopie-structure. The present report summarises these recent developments and their potential relevance for arrhythmogenesis. PMID- 9202852 TI - Glucose metabolism and protective biochemical mechanisms in a rat brain affected by kaolin-induced hydrocephalus. AB - To clarify glucose metabolism in a hydrocephalic rat brain, substances related to glycolytic metabolism were biochemically measured. Kaolin-induced hydrocephalic rats were sacrificed and lactate dehydrogenase (LDH), LDH isozyme, lactate, adenosine triphosphate (ATP), and isocitrate dehydrogenase (ICDH) were measured in the following regions: cortex, thalamus, midbrain, hippocampus, cerebellum, and pons with medulla. During the development of hydrocephalus, lactate and LDH increased in most regions, the LDH M-subunit increased in the cortex, and ICDH decreased in most regions. However, ATP levels did not change. The increases in lactate, LDH and M-subunit suggested an anaerobic environment in the cell leading to activation of the anaerobic glycolysis. The decrease in ICDH represented a diminution of the tricarboxylic acid cycle. Through these alterations, the ATP level can be kept constant during the course of hydrocephalus, allowing the brain to create a better biochemical milieu. PMID- 9202853 TI - A rat model of spontaneously arrested hydrocephalus. A behavioural study. AB - HTX rats with congenital hydrocephalus that survived for more than 2 months are termed spontaneously arrested hydrocephalic rats. These rats showed impairment in learning, a reverse light-dark discrimination task in a Y-maze. A clear relationship between learning impairment and ventricular dilatation was observed in spontaneously arrested hydrocephalic animals. The usefulness and limitations of HTX-rats as an animal model of spontaneously arrested hydrocephalus are discussed. PMID- 9202854 TI - Surgical treatment of syringomyelia associated with spinal dysraphism. AB - Clinical and radiological features of syringomyelia in 15 patients with spinal dysraphism are reported. There were 8 patients with occult spinal dysraphism (lumbosacral lipoma) and 7 with spina bifida aperta (meningomyelocele). Syringomyelia with spinal dysraphism can be radiologically divided into two types according to the dysraphic state. The syrinx in the patients with occult spinal dysraphism occurred immediately rostral to the lipoma and was localized to the lower thoracic to lumbar levels, while in the meningomyelocele patients the syrinx extended from the cervical to the thoracic level. Large syrinx formation was recognized in 1 of the 7 occult spinal dysraphism cases and 3 of the 8 meningomyelocele cases. For syringomyelia with occult spinal dysraphism, 4 patients underwent syringo-subarachnoid shunting (S-S shunt, 2 cases) or syringostomy (2 cases) during an untethering operation. In the case of meningomyelocele, S-S shunts were placed in 2 patients. Collapse of the syrinx was achieved in all 6 patients who underwent S-S shunting or syringostomy. Decreased size of the syrinx was also noted in 3 occult spinal dysraphism patients who underwent untethering operations alone. In conclusion, a large syrinx in the case of spinal dysraphism should be surgically treated. S-S shunting is effective in both types of syringomyelia. Foramen magnum decompression may be an alternative method of surgical treatment for syringomyelia in patients with meningomyelocele. PMID- 9202855 TI - Disc protrusion in the child. Particular features and comparison with neoplasms. AB - Lumbar intervertebral disc herniation, although common in adults, is infrequent in the young, and especially in patients under 17 years old. In this work we review clinical data pertaining to two pediatric groups of patients whose main complaint was low back pain and/or sciatica, trying to identify factors that might contribute to their earlier referral and to the differential diagnosis of protruded disc and spinal neoplasm in this population. Group A comprises 17 youngsters diagnosed as having lumbar herniated nucleus pulposus and group B, 16 children with neoplasms of the lower thoracic and lumbosacral regions. Both groups were similar in sex distribution and symptoms of pain and numbness. However, there was a striking difference in age at presentation. No patient in group A was younger than 11 years, while most of those in group B were in their first decade of life (P = 0.018). The classic clinical onset in the children with herniated discs started with low back pain and sciatica, as in the children with neoplasms, although in subgroup B leg pain tended to be bilateral. The usual examination findings in both groups were spinal rigidity and sensory loss, but motor weakness and impaired reflexes were found to be more frequent in the group with spinal growths (P = 0.02). Children with lumbosacral neoplasms also tended to present with atypical symptoms (acute onset, intracranial hypertension, subarachnoid hemorrhage and abdominal pain), while this was the exception in the group with herniated discs. Plain radiographs of the pediatric spine showed that X-ray examination is still a good tool for diagnosing spinal growths compared with their scant utility in disc herniations (P = 0.001). During the survey we were impressed by the children's apparent good tolerance to pain, which is probably due to the lack of the emotional component of pain in adults and explains their delayed referral for neurosurgical consultation. However, all modalities of treatment seemed to be effective in children, chemonucleolysis and surgery being extraordinarily effective in this age group. Accordingly, we see no reason for long-term conservative therapy in children with lumbar and sciatic pain; on the contrary, we believe these patients should be offered earlier neurosurgical treatment. PMID- 9202856 TI - Early detection of neurological manifestations in achondroplasia. AB - Achondroplasia (ACh) is the most frequent bone dysplasia. The mode of inheritance is autosomal dominant. The incident of neurological complications ranges between 20% and 47%; frequently the symptoms are subtle but are due to such serious conditions as cervicomedullary compressive syndromes, syringomyelia or hydrocephalus; thus, the early identification of this disorder is very important. We made a prospective study of 39 patients (20 female, 19 male) with ACh; their ages ranged from 3 months to 17 years (mean 4 years and 6 months). All patients had hypotonia and psychomotor delay; 3 had recurrent apnea, 1 developed radicular syndrome and 1 had leg paresthesias. The CT scan was normal in 5, 20 had cortical atrophy and 18 communicating hydrocephalus; we identified foramen magnum abnormalities in 28 patients, and reduced craniocervical junction with cervicomedullary compression in 6. Myelography and myelotomography demonstrated spinal compression in 12 patients. The MRI showed cervicomedullary infarct in 1, syringomyelia in 2 and diastematomyelia in 1. The somatosensory evoked responses (SSER) were very useful in the early identification of brain stem and spinal abnormalities. We concluded that the neurological manifestations of pediatric patients with ACh are frequent and very important, demanding comprehensive clinical evaluation even in asymptomatic patients, especially those with severe hypotonia or SSER alterations. PMID- 9202857 TI - Hemodynamic compromise as a factor in clinical progression of Sturge-Weber syndrome. AB - We divided eight patients with Sturge-Weber syndrome into two groups, depending on the presence or absence of clinical progression, and investigated differences in brain hemodynamics between the groups by measuring regional cerebral blood flow (rCBF) before and after acetazolamide activation using stable xenon computed tomography. The most evident difference between the groups was acetazolamide vaso reactivity (delta CBF) in the primarily healthy area remote from lesion. delta CBF on the contralateral side and in distant areas on the affected side was significantly lower in the group with progression group (unpaired t-test), although both groups had similar delta CBF values in the area of primary lesion. This remote hemodynamic compromise might aggravate the clinical condition. Operations were performed on three patients with progressive disease, resulting in good seizure control with anticonvulsants and a variable degree of improvement in neurological symptoms and vasoreactivity. The neurological improvement appeared to correlate with the size of the resected area and the increase in delta CBF after surgery. PMID- 9202858 TI - Multiple EDAS (encephalo-duro-arterio-synangiosis). Additional EDAS using the frontal branch of the superficial temporal artery (STA) and the occipital artery for pediatric moyamoya patients in whom EDAS using the parietal branch of STA was insufficient. AB - Although parietal EDAS or STA-MCA anastomosis are effective in pediatric moyamoya disease, they do not adequately prevent ischemia in the frontal and occipital lobes. Some additional methods that can prevent ischemia in the frontal and occipital lobes are sometimes needed. We investigated whether EDAS using a frontal branch of the superficial temporal artery (frontal EDAS) or EDAS using the occipital artery (occipital EDAS) is preferable. Frontal or occipital EDAS was performed at 15 sites in seven patients with pediatric moyamoya disease. The outcome was estimated by angiography 3 months later, CT findings 3 months later, neurological findings during the follow up period and perioperative complications. The mean follow up period was 14 +/- 6 months after frontal or occipital EDAS. As results, good revascularization from frontal or occipital EDAS was shown in ten of fourteen surgical sites (71%) in angiography. None of the patients showed deterioration of symptoms after frontal or occipital EDAS during the follow up period. None of the patients developed surgical complications. In conclusion, multiple EDAS using the frontal branch of STA and the occipital artery is an effective and safe method for preventing ischemia in the frontal and occipital lobe in pediatric moyamoya disease. PMID- 9202859 TI - Immature teratoma producing alpha-fetoprotein without components of yolk sac tumor in the pineal region. AB - A case of pineal region tumor in a 9-year-old boy with a high serum alpha fetoprotein (AFP) level is reported. The serum levels of beta-human chorionic gonadotropin (HCG) and placental alkaline phosphatase (PLAP) were not elevated. The tumor was composed of radiologically different components and was removed surgically. Postoperative radiation therapy was performed and the serum level of AFP gradually declined to the normal range. The pathological diagnosis was immature teratoma, and no elements of yolk sac tumor or embryonal carcinoma were found. In the immunohistochemical study, AFP was detected in the cytoplasm of gastrointestinal-type epithelium and primitive neuroepithelial element. This is considered a rare case of intracranial immature teratoma in which AFP was detected immunohistochemically in the columnar epithelium and immature neural tissue. PMID- 9202861 TI - Bobble-head doll syndrome associated with subduroperitoneal shunt malfunction. AB - Bobble-head doll syndrome is known to be associated with aqueductal stenosis or cystic lesions of the III ventricle. The direction of movement is usually vertical. In the literature, only five cases of purely horizontal movement have been reported. Bobble-head doll syndrome manifested as a sign of shunt malfunction has been described in one case with a ventriculoperitoneal shunt. The authors report on a 10-year-old boy who showed subduroperitoneal shunt malfunction associated with horizontal bobble-head doll syndrome. The head bobbing disappeared immediately after shunt revision. Unlike the previously reported cases, in the present case the lesion was asymmetric, though the significance of this for the lateral movement is not clear. This case also showed more marked ventricular dilatation on subduroperitoneal shunt malfunction than in the pre-shunt state. The underlying mechanism of the ventricular dilatation is unknown. PMID- 9202860 TI - Chiasmal gliomas with spontaneous regression: proliferation and apoptosis. AB - Spontaneous regression of chiasmal gliomas without associated neurofibromatosis has been occasionally described. In this paper we present two patients with chiasmal glioma whose tumors decreased in size during the postoperative course. Neither patient received radiotherapy or chemotherapy. We examined the proliferative activity of the excised tumors by determining the Ki-67 labeling index and searched for apoptotic cells using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) method. The Ki-67 labeling index of the tumor of case 1 was 0.63%, and apoptotic cells were detected in some areas. In case 2, the Ki-67 labeling index was 19.5% and a large number of apoptotic cells were evident. As estimated from the respective apoptosis data, our results would indicate that tumor regression may occur when the rate of cell loss is greater than that of tumor growth. PMID- 9202862 TI - The cardio-facio-cutaneous syndrome: a long-term follow-up of two patients, with special reference to the neurological features. AB - The cardio-facio-cutaneous (CFC) syndrome is a newly recognized syndrome characterized by congenital heart defects, a particular facial appearance and abnormalities of hair and skin; apart from these, the most evident signs are mental retardation and other neurological abnormalities. Some of the phenotypic manifestations resemble those of Noonan syndrome, but this involves different neurological problems. We report on two children affected by cardio-facio cutaneous syndrome who have been followed up for at least 6 years. The psychomotor retardation was severe. During the follow-up period we made a careful analysis of the neurological aspects of the course in these two patients and wish to stress that some neurological features are peculiar to this syndrome and can be useful in the diagnosis. PMID- 9202863 TI - Variations in living donor graft rates by dialysis clinic: effect on outcome and cost of chronic renal failure therapy. AB - OBJECT: Examination of nephrology practice variations in living donor renal grafts to determine their influence on organ supply, quality, and cost of chronic renal failure therapy. MATERIALS: Saskatchewan chronic dialysis, cadaveric, and living donor renal grafts in 1983-1994 inclusive. RESULTS: Saskatchewan has three dialysis (I, II, III) and one transplant clinic. In the period the renal graft incidences/million population by these dialysis clinics by organ source were; Cadaveric: 23.1, 23.2, 21.1 (p = ns). Living: 5.4, 21.7, 8.3 (I or III vs II p < 0.000, I vs III p < 0.061). Total: 28.7, 44.7, 29.4. Living donor series A is 79 grafts in patients under age 60 with primary renal disease. Series B is 20 grafts in patients with secondary renal disease or over age 59. Series A ten-year actuarial patient survival is 92% and B 44%. Series A ten-year actuarial graft survival (including regrafts) is 77% and B 39%. Rehabilitation rate in patients with functioning grafts is 88.5%. Province-wide extension of the Clinic II living donor graft rate in 1983-1994 would have produced 160 more renal grafts or 59% of those receiving chronic dialysis in 1994. The annual maintenance for a graft with the initial grafting cost taken over five years was $10,825 and the dialysis cost $40,100. CONCLUSIONS: (1) nephrology practice variations caused a 2.5-4.0-fold difference in living donor renal graft rates, indicating patient education by the attending nephrologist influences the living donor transplantation rate, (2) with such education the combined living donor and the cadaveric organ supply virtually meets graft demand, (3) living donor renal grafts yield a better quantity and quality of life and better cost control than dialysis with their annual cost being one-quarter that for dialysis. PMID- 9202865 TI - Risk factors for left ventricular hypertrophy in chronic hemodialysis patients. AB - Left ventricular hypertrophy is reported to be common in chronic hemodialysis patients, and also to increase the risk for mortality in chronic hemodialysis patients. An echocardiographical and clinical study was carried out to investigate the risk factors for left ventricular hypertrophy in 151 non-diabetic chronic hemodialysis patients without valvular diseases or myocardial infarction in two hemodialysis units in Fukuoka, Japan. The left ventricular mass index (LVMI) correlated positively to age, systolic blood pressure and interdialysis weight gain while it correlated negatively to the duration of hemodialysis therapy and hematocrit. Resorting to a multivariate analysis, the LVMI was found to positively correlate to age and the systolic blood pressure while it correlated negatively with the duration of hemodialysis therapy and the hematocrit level. These findings suggest that hypertension and anemia may thus be independent risk factors for left ventricular hypertrophy. PMID- 9202864 TI - Vesicoureteral reflux after kidney transplantation: clinical significance in the medium to long-term. AB - 103 patients who received a cyclosporine-treated primary cadaver kidney transplant (TX) at our center between 1985 and 1989, whose graft survived for more than 1 year and who accepted to undergo voiding cystography after TX were analyzed and grouped according to the highest grade (regardless to whether active or passive) of vesicourteral reflux (VUR): group 0, absent (n = 14); group 1-2, grade I or II (n = 62); group 3, grade III (n = 27). Patient follow-up ranged from 5 to 10 (median 7) years. Patient and graft survivals and prevalence of hypertension (defined as the persistent need of antihypertensive therapy), did not differ significantly between groups (Mantel-Cox test p: n.s. in all cases). GFR (Cockroft and Gault) and proteinuria were evaluated with ANOVA for repeated measures at 1, 2, 3, 4 and 5 years in the 96 patients (group 0: 13, group 1-2: 56, group 3: 27) whose grafts lasted for 5 years or more. Neither GFR values (p: n.s.) nor GFR behaviour over time (p: n.s.) differed between groups, although a progressive decline of GFR was noted in all groups (p < 0.002). Proteinuria neither showed any significant differences between groups in values (p: n.s.) or behaviour over time (p: n.s.), nor any trend in behaviour over time in all groups as a whole (p: n.s.). Finally, in the first 5 years after TX the 3 groups did not differ for number of urinary tract infections (UTIs) (mean value for all patients: 2.5, range 0-22, episodes/pt/5 years) (p: n.s.), or for number of UTIs with leukocyturia (mean 0.6, range 0-6, episodes/pt/5 years) (p: n.s.), or for number of febrile UTIs (mean 0.3, range 0-5, episodes/pt/5 years) (p: n.s.), or for number of UTIs with sepsis (mean 0.1, range 0-2, episodes/pt/5 years) (p: n.s.). The same results were obtained when, instead of episodes/ pt/5 years, percentages of patients without or with 1 or more of such episodes in the same period were considered. In conclusion, VUR does not seem to be hazardous for the transplanted kidney in the medium to long-term. PMID- 9202866 TI - Hepatitis C virus genotypes in hemodialyzed patients: a multicentric study. AB - The few studies concerning HCV genotypes in hemodialyzed patients concerned small groups of patients, issued from single units. Using the RFLP typing method in the 5' non-coding region (5' NCR), we performed the genotyping of HCV strains of 80 patients issued from 14 Belgian dialysis units. Forty-seven of the 80 patients were also tested by Inno-Lipa II (Innogenetics, Gent, Belgium). Sixty-four of 80 patients (80%) were infected with subtype 1b, 2 (2.5%) with subtype 1a, 6 (7.5%) with subtype 2a and 1 (1%) with subtype 2b; 6 patients (7.5%) showed a type 4 and 2 patients (2.5%) a type 5 infection, respectively. Only 1 patient (1%) showed a mixed infection (1a and 1b). In the 47 patients tested by both methods, we observed a very good agreement (98%) between RFLP and Inno-Lipa II's results. Our multicentric study detected HCV genotypes 4 or 5 in 8/80 (10%) of hemodialyzed patients in Belgium. The substantial prevalence of these genotypes could not be fully explained by a nosocomial spread of HCV infection: indeed, four patients belonged to dialysis units located in different cities, and two appeared infected with distinct subtypes in a same unit. Thus, the discovery of a "rare" HCV genotype in several hemodialyzed patients does not always point to nosocomial HCV transmission. PMID- 9202867 TI - The effect of propionyl L-carnitine on skeletal muscle metabolism in renal failure. AB - The effect of propionyl L-carnitine on skeletal muscle metabolism in chronic renal failure. Carnitine deficiency, resulting in defective oxidative ATP synthesis, has been implicated in the myopathy of chronic renal failure. Using 31P magnetic resonance spectroscopy we examined calf muscle metabolism in 10 dialysed patients before and after 8 weeks of propionyl L-carnitine (PLC) 2 g.p.o. daily. Resting phosphocreatine/ATP (4.41 +/- 0.20 [SEM]) decreased to normal control levels on PLC (3.98 +/- 0.14; controls 4.00 +/- 0.06). In contrast, there was no effect of PLC on aerobic and anaerobic metabolism of muscle during or following 2-10 min exercise. The maximal calculated oxidative capacity (Qmax) remained below normal (28 +/- 3 mM/min before and 24 +/- 3 mM/min after PLC; controls 49 +/- 3 mM/min). Qmax correlated positively with hemoglobin concentration ([Hb]) after PLC (p < 0.03). Oxidative capacity assessed by phosphocreatine recovery T significantly improved with PLC administration (0.93 +/- 0.1 to 0.74 +/- 0.08 min) in those patients (n = 6) with [Hb] > 10 g/dl. [Hb] was rate limiting to oxidative metabolism in recovery from exercise but only following treatment with PLC. Patients with anemia or those subjects who use relatively more non-oxidatively synthesized ATP during exercise, do not respond to PLC. Oxidative metabolism did not normalize on PLC suggesting that anemia and carnitine deficiency are not the only causes of mitochondrial dysfunction in renal failure. PMID- 9202868 TI - Kinetics of phosphorus during hemodialysis and the calculation of its effective dialysis clearance. AB - Removal of Pi from the body during hemodialysis (HD) is more difficult than that of urea or creatinine. Therefore, the precise evaluation of its effective dialysis clearance (K) is of practical significance. Estimation of K of Pi is associated with some problems because its kinetics during HD cannot be expressed by a simple mathematical function. The purpose of this study was to determine to what extent the calculation of K of Pi is affected by the calculation method employed. In six patients, the kinetics of Pi in serum and effluent dialysis fluid was repeatedly monitored during HD. The first stage of HD, associated with a fairly rapid decrease in serum Pi concentrations, was followed by their stabilization at a constant level. The average predialysis serum Pi was 2.69 (+/ 0.41) mmol/l. Three hours after the start of HD, the average serum Pi concentration was 1.33 (+/-0.33) mmol/l and, four hours after the start of HD, it was 1.34 (+/-0.30) mmol/l in average. The following procedures were used for K calculation: (a) the amount of Pi removed during the time interval at which serum Pi concentration was stabilized at a constant level (and calculated per one minute) was divided by the recorded stabilized serum value, (b) the amount of Pi removed from the body during HD and calculated per one minute (D/t) was divided by the average value of pre- and post-HD serum concentrations, (c) K calculation assumed exponential function of the decrease in serum Pi during HD, (d) the total amount of Pi removed during HD was divided by the area under the curve of serum concentrations (D/AUCs), (e) the amount removed during HD and calculated per one minute (D/t) was divided by the serum concentration recorded at the midpoint of HD. The average value of K of Pi calculated on the basis of method (a) was 122.5 (+/-33.6) ml/min and was significantly higher (p < 0.01) than that calculated on the basis of methods (b) (95.7 +/- 31.2 mmol/l) and (c) (103.2 +/- 31.9 mmol/l), but was significantly lower (p < 0.05) than that calculated according to method (e) (142.8 +/- 46.3 ml/min). The average value of K calculated on the basis of method (a) did not differ significantly from that calculated according to method (d) (124.7 +/- 35.2 ml/min). Calculation of K according to method (a) is identical with the classic method for calculating the renal clearance of various substances and used generally in clinical nephrology. We therefore used this method as a reference one. The results obtained suggest that, of the other methods used for K calculation, only method (d) based on the calculation of the D/AUCs ratio gives values which do not differ significantly from those obtained using method (a). PMID- 9202869 TI - Fluoroscopic manipulation of Tenckhoff catheters: outcome analysis. AB - Wire manipulation under fluoroscopic control can correct malposition of peritoneal catheters in a fast and safe manner. This study was performed to evaluate the success rate of wire manipulation and identify factors that may predict outcome. MATERIAL AND METHODS: Data were prospectively collected between January 1, 1986 and June 30, 1996 among all patients with peritoneal catheter malfunction requiring manipulation at a single center. Manipulations were performed using a flexible guide wire under aseptic fluoroscopic control. All patients had flat plates of the abdomen pre and post manipulation and received antibiotics after the procedure. The direction of the subcutaneous tunnel at the point of entry into the peritoneal cavity was calculated from the X-rays by measuring the angle formed by the horizontal and the distal subcutaneous tunnel. Success was defined as adequate peritoneal catheter function at three months. The success rate was correlated with type of catheter, patient weight and the subcutaneous tunnel orientation. RESULTS: 1250 Tenckhoff double-cuff peritoneal catheters were inserted and 69 (5.5%) were manipulated (59 straight and 10 curled). The median for time elapsed between peritoneal catheter insertion and wire manipulation was 18 days (range 1 day-5 years). The overall success rate was 60.9% (61% for straight and 60% for curled peritoneal catheters). The mean patient weight for successes was 71.4 +/- 12.4 and 84.0 +/- 17.2 kg for failures (p < 0.005). Subcutaneous tunnel orientation between 30 and 120 degrees was associated with the highest success and those beyond 120 degrees with the highest failure rate. No complications were observed. CONCLUSIONS: Wire manipulation under fluoroscopic control of Tenckhoff peritoneal catheter in the treatment of malfunction is a safe and highly effective procedure. Obesity and cephalad orientation of the subcutaneous tunnel were associated with less favorable outcome. PMID- 9202870 TI - Is CAPD an effective treatment for ESRD patients with a weight over 80 kg? AB - We studied the effectiveness of CAPD in large patients (> 80 kg) (group B, n = 49) by comparing them to a group of patients whose body weight was 60-80 kg (group A, n = 193). Patients in group B were two years younger (55.4 versus 57.7 years, p < 0.01), were predominantly male (M: F ratio 33/16 vs 84/109) and had slightly higher residual creatinine clearance (8 ml/min vs 6 ml/min) at the beginning of treatment. The prevalence of diabetes and the prevalence of comorbid conditions in the two groups were similar. The incidence of peritonitis was similar between the two groups. Patients with a large weight spent significantly fewer days in hospital (20.6 +/- 25 vs 23.4 +/- 35.0 days/year); reasons for hospitalization were similar, except for weakness/fatigue that was more frequent (10%) in group B than in group A (2%). The initial weekly dialysate volume was similar in the two groups (57 +/- 51 in group B and 56 +/- 101 in group A) and increased in both groups at the end of the study to 60 +/- 141 in group B and 61 +/- 171/week in group A. The weight of 6 patients in group B and 5 in group A decreased below the range of that group. On the contrary the weight of 28 patients in group A increased to the range of group B. Based on the final weight there were 166 patients whose weight was 60-80 kg, and 71 patients whose weight was over 80 kg (80-109 kg). Patient survival was similar between the two groups. There was a significantly higher death rate among those patients whose weight decreased in both groups compared to those whose weight increased or remained stable. We conclude that CAPD is an effective treatment in the management of ESRD patients with weights over 80 kg. There is a high mortality among patients whose weight decreases irrespective of their initial weight. PMID- 9202871 TI - Isolated cerebral aspergilloma--long-term survival of a renal transplant recipient. AB - A renal transplant recipient with isolated cerebral aspergilloma 4 months after allograft transplantation is reported. On admission cerebral computed tomography showed a ring-enhancing mass in the left frontal hemisphere and aspirated purulent material revealed A. fumigatus hyphae. He was cured by short-term antifungal therapy and neurosurgical removal of the well demarcated lesion. He is still alive more than two years later and the renal transplant is well functioning. This is the first report of a renal transplant recipient with isolated cerebral aspergillosis without any relapse and only the third patient who has survived longer than 3 months. Early diagnostic procedures with rapid confirmation of aspergillus infection are pivotal for a benign clinical course. PMID- 9202872 TI - Quinine-induced hemolytic uremic syndrome. AB - Although quinine use is very common, hemolytic uremic syndrome (HUS) following exposure to quinine is only a recently reported phenomenon, with the first description published in 1991. Previous reports have concentrated on the nature of the hematological process and in particular characterization of the quinine induced antibodies involved. We present a case of HUS with a clear temporal and immunological relationship to quinine which demonstrates the pathognomonic renal features of HUS. An indirect antiglobulin test with the patient's serum agglutinated red blood cells only in the presence of quinine. Renal biopsy features included glomerular and arteriolar endothelial swelling, capillary loop thrombi, mesangiolysis, segmental sclerosis and segmental ischemia. Early empiric treatment with plasma exchange and corticosteroids was instituted and this resulted in recovery of renal function to normal. PMID- 9202873 TI - Cerebral involvement in two patients with Wegener's granulomatosis. AB - Two cases of cerebral involvement in Wegener's granulomatosis (WG) are described. The course of the disease in both patients was characterized by sudden onset and fatal outcome, despite maximum immunosuppressive therapy. Cerebral involvement is a rare complication of WG. Over the past two decades, only a small number of case reports appeared of patients with WG who showed this complication. Since the era of cyclophosphamide therapy, it is commonly assumed that cerebral involvement in WG has no influence on patient survival. However, the two patients described here both died shortly after the occurrence of central neurological symptoms. PMID- 9202874 TI - Renal artery stenosis complicating Cogan's syndrome. PMID- 9202875 TI - Comment to Sunder-Plassmann and Horl. PMID- 9202876 TI - Catatonia and the neuroleptics: psychobiologic significance of remote and recent findings. AB - The previously common occurrence of catatonic schizophrenia and catatonic symptoms among schizophrenic patients has diminished sharply; catatonic symptoms now occur more frequently in association with severe affective disorders or with general medical conditions. Catatonia is generally viewed as a peculiar and puzzling syndrome and attracts limited attention. Yet significant catatonic symptoms tend to be present in close to 10% of patients admitted to psychiatric inpatient facilities. The dynamic significance of catatonia can be recognized by considering the original biologic role of catatonia in schizophrenia as an opposite to the paranoid disorder. Szondi viewed catatonia as an attempt at self healing of the paranoid psychosis with its threatening total expansion, by extreme constriction of the ego. The previously predominant primary association of catatonia with schizophrenia has been eclipsed as neuroleptics have supplanted the endogenous self-healing attempt of catatonia, preventing the occurrence of catatonic symptoms in schizophrenia. Neuroleptics in fact duplicate or approximate the symptoms of catatonia by producing mental immobilization, hypokinesis (parkinsonism and dystonia), hyperkinesis (akathisia), and pernicious catatonia in the modern guise of the neuroleptic malignant syndrome (NMS). Patients with past or present catatonic symptoms are particularly vulnerable to NMS, and treatment of catatonia requires avoidance of neuroleptics and the use of benzodiazepines or electroconvulsive therapy (ECT). The extreme negativism and constriction of consciousness in catatonia suggest a primary role of the frontal lobes, with secondary involvement of the extrapyramidal system and its movement disorders. In an attempt to integrate clinical, psychologic, neuropharmacologic, and neurochemical findings, a modern dynamic neuropsychiatry must appreciate the major significance of catatonia. PMID- 9202877 TI - Gender differences in DSM-IV alcohol use disorders and major depression as distributed in the general population: clinical implications. AB - This study examined gender differences within and between five groups of subjects drawn from a large representative sample of the United States population and classified as having either major depression (MDD) only, alcohol use disorder (AUD) only, or primary, secondary, or concurrent depression to determine if these diagnostic profiles (1) were consistent with those drawn on clinical samples and (2) might suggest potential clinical implications. Respondents (N = 9,985) from a nationally representative survey of the United States population met DSM-IV criteria for classification into these five mutually exclusive groups that were compared within and between groups by gender on the characteristics of each disorder. The results were consistent with those of other studies: (1) gender distributions of AUD and depressive disorder remain almost mirror opposites, and (2) comorbid disorders are more severe than either of the conditions appearing singly. Findings of particular interest were that the synergistic effects of an alcohol and a depressive condition operate equally for both men and women with concurrent depression. This points to the necessity of attending carefully to gender biases when dealing with comorbid conditions, last we fail to take alcoholism in the presence of depression seriously enough in women and vice versa in men. Additionally, women with primary depression are at high risk for suicide and thus may require special attention in the evaluative phase of treatment. PMID- 9202878 TI - Age effects on the clinical presentation of alcoholics at a psychiatric hospital. AB - Little is known about the effects of age on the clinical presentation of alcoholism in various treatment settings, despite the clinical importance of this factor. This study evaluates the effects of age on the clinical profile of 604 alcoholics who presented for initial evaluation and treatment at a psychiatric hospital. Young alcoholics displayed the most prominent substance use, antisocial behavior, depressive symptoms (including suicidality), and impulsivity. Early middle-aged alcoholics displayed the highest levels of drinking. Elderly alcoholics displayed the highest levels of cognitive dysfunction, although some level of cognitive dysfunction was present among even the youngest alcoholics. These findings confirm and clarify the effects of age on the clinical profile of alcoholics presenting for initial evaluation at a psychiatric hospital. PMID- 9202880 TI - Violence in psychiatric patients: the role of psychosis, frontal lobe impairment, and ward turmoil. AB - The purpose of the study was to identify psychiatric symptoms, neurological impairments, and situational factors associated with the emergence of violence and with its persistence. Psychiatric symptoms were assessed in newly admitted physically assaultive psychiatric patients and nonviolent controls. Patients were than evaluated for 4 weeks to determine the persistence or resolution of these physical assaults. Patients who showed marked resolution of assaults were classified as transiently violent (n = 41), and those who remained assaultive throughout were categorized as persistently violent (n = 34). At the end of 4 weeks, all patients received a comprehensive psychiatric and neurological assessment. Physical assaults were associated initially with prominent positive psychotic symptoms. Both transiently and persistently violent patients were more psychotic than the nonviolent controls; however transiently violent patients showed better resolution of these symptoms over the 4 weeks. They also evidenced less frontal lobe impairment on the neurological examination than the persistently violent patients. The two violent groups differed in their susceptibility to environmental influences: the surrounding ward agitation fostered physical assaults in transiently but not in persistently violent patients. This differentiation between transiently and persistently violent patients has major implications for the comprehensive treatment of violent behavior. PMID- 9202879 TI - Characteristics of injection drug users derived from a large family study of alcoholism. AB - Most descriptive studies on injection drug users (IDUs) has used treatment or referral samples. This study uses relatives ascertained through the Collaborative Study on the Genetics of Alcoholism (COGA) to describe patterns of drug use, psychiatric comorbidity, and selected high-risk behaviors among IDUs not ascertained through treatment or other referral networks. Relatives (N = 5,520) of alcoholic probands were administered a semistructured interview, the Semistructured Assessment for the Genetics of Alcoholism (SSAGA), which assesses lifetime psychiatric symptoms and psychoactive substance use. IDUs were compared with these who had never used cannabis more than 20 times or other drugs more than 10 times (minimal drug use group), those who had used cannabis more than 20 times but no other illicit drugs more than 18 times (cannabis users), and those who had used other drugs more than 10 times but who had never injected (other drug users). Compared with other drug users, IDUs reported using more classes of drugs and began drug use at an earlier age. IDUs were significantly more likely to receive diagnoses of antisocial personality disorder (ASPD) and alcohol dependence, and reported elevated rates of suicidal ideas and attempts. IDUs were more likely to report a variety of behavioral difficulties beginning before age 15, and were more likely to engage in high-risk sexual behaviors. Half of the IDUs reported having shared needles; shares were more likely to receive a diagnosis of ASPD, but did not differ on reporting high-risk behaviors. IDUs, regardless of whether selected through treatment, have high lifetime rates of mood disturbance and ASPD, and are likely to have a history of conduct difficulties beginning before age 15 years and to subsequently engage in a variety of other high-risk behaviors. PMID- 9202881 TI - Antisocial and borderline personality disorders: two separate diagnoses or two aspects of the same psychopathology? AB - Antisocial personality disorder (ASPD) and borderline personality disorder (BPD) have a number of points of overlap: in symptoms, in personality dimensions that underlie their phenomenology, in community prevalence, in risk factors, and in outcome and response to treatment. Both disorders have a common base in impulsive personality traits, but the behavioral differences between them are shaped by gender. Further research is suggested to explore the commonalities and differences between ASPD and BPD. PMID- 9202882 TI - Validation of the structured interview for disorders of extreme stress. AB - There is a dearth of valid instruments to evaluate the primary sequelae of sexual abuse. The present study examined the construct validity of the subscales of the Structured interview for Disorders of Extreme Stress (SIDES), a measures of the associated features of posttraumatic stress disorder (PTSD). Using a clinical sample of 74 survivors of childhood sexual abuse with PTSD, this study demonstrated that the subscales of the SIDES (alterations in regulation of affect and impulses, alterations in attention or consciousness, alterations in self perception, and somatization) correlated highly with instruments hypothesized to measure similar constructs. Divergent validity was established for each of four subscales of the SIDES. Findings from this study and other research on the psychometrics of the SIDES indicate that it is a valid measure of the associated features of PTSD in survivors of childhood sexual abuse. PMID- 9202883 TI - Regulatory functions of phospholipase A2. AB - Phospholipase A2 (PLA2) plays crucial roles in diverse cellular responses, including phospholipid digestion and metabolism, host defense and signal transduction. PLA2 provides precursors for generation of eicosanoids, such as prostaglandins (PGa) and leukotrienes (LTs), when the cleaved fatty acid is arachidonic acid, platelet-activating factor (PAF) when the sn-1 position of the phosphatidylcholine contains an alkyl ether linkage and some bioactive lysophospholipids, such as lysophosphatidic acid (lysoPA). As overproduction of these lipid mediators causes inflammation and tissue disorders, it is extremely important to understand the mechanisms regulating the expression and functions of PLA2. Recent advances in molecular and cellular biology have enabled us to understand the molecular nature, possible function, and regulation of a variety of PLA2 isozymes. Mammalian tissues and cells generally contain more than one enzyme, each of which is regulated independently and exerts distinct functions. Here we classify mammalian PLA2s into there large groups, namely, secretory (sPLA2), cytosolic (cPLA2), and Ca(2+)-independent PLA2s, on the basis of their enzymatic properties and structures and focus on the general understanding of the possible regulatory functions of each PLA2 isozyme. In particular, the roles of type II sPLA2 and cPLA2 in lipid mediator generation are discussed. PMID- 9202884 TI - Major histocompatibility complex (MHC) class I recognition by natural killer cells. AB - NK cells express clonally distributed receptors specific for MHC class I molecules. Structurally, these receptors belong to the C-type lectin superfamily in mouse and to the immunoglobulin superfamily in human. Functionally, they can be distinguished as inhibitory or stimulatory. Inhibitory receptors block NK cell mediated cytotoxicity upon binding to HLA class I ligands. This function is mediated by phosphorylation of cytoplasmic tyrosines, which recruit the protein tyrosine phosphatase SHP-1. Stimulatory receptors also bind HLA class I, lack cytoplasmic tyrosine-based motifs, and trigger NK cell-mediated cytotoxicity. All these receptors are characterized by a limited diversity allowing for sensitive detection of loss of MHC class I molecules on autologous transformed and virally infected cells. PMID- 9202885 TI - Signals for activation of the GM-CSF promoter and enhancer in T cells. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is one of the many cytokines produced following T-cell activation. It is also produced in a variety of other cell types, in particular following activation by inflammatory mediators. Changes in the rate of transcription are important in the control of GM-CSF expression in T cells and in fibroblasts and endothelial cells. The GM-CSF gene contains two distinct transcriptional control regions. These are the proximal promoter consisting of the first 120 bp from the transcription start site and an enhancer located approximately 3 kb upstream from the proximal promoter. Distinct regions of the proximal promoter respond to a wide array of signals such as phorbol myristate acetate (PMA) and Ca2+ ionophore or phytohemaglutinin (PHA), CD28 activation, human T leukemia virus (HTLV)-1 tax, TNF, and interleukin 1 (IL-1). The transcription factors that mediate these responses have mainly been defined, with the major inducible proteins being the NF-kappa B/rel and AP-I families of transcription factors. In contrast to the promoter, the enhancer responds only to PMA and Ca2+ ionophore signals and binds NFAT/AP-1 complexes that appear to mediate its function. PMID- 9202886 TI - Regulation of the function of eosinophils and basophils. AB - Both eosinophils and basophils play active pathogenic roles in the inflammation associated with allergic disorders. Both types of cells share a majority of their cell surface structures, and because of these common surface molecules, both cells can be stimulated with a single ligand simultaneously. The growth of both types of cells is controlled by IL-3, GM-CSF, and IL-5. All three growth factors are also capable of priming both eosinophils and basophils for enhanced biological functions, such as increased mediator release and prolonged survival. Both cells express beta 2 integrins, and in contrast to neutrophils, they also express several beta 1 integrins. Ligation of these adhesion molecules also transduces the intracellular signal leading to regulation of the cellular functions. In this review, we briefly describe the effects of various ligands of surface receptors and several pharmacological compounds on the functions of human eosinophils and basophils. PMID- 9202887 TI - Biochemistry of antigen receptor signaling in mature and developing B lymphocytes. AB - The function of the surface antigen receptor of B cells (BCR) has been extensively studied with respect to the activation of mature B lymphocytes. B cells at other points of development (e.g., pre-B cells, immature B cells, and germinal center B cells) also express forms of the BCR, and for cells at these developmental timepoints, signaling through the BCR in some cases may lead to outcomes other than B-cell activation. Understanding the molecular events that are initiated by BCR crosslinking would enhance our understanding of the regulation and functional results of BCR signaling throughout B-cell development. In this article we review the current understanding of BCR signal transduction from initiation of the signal through changes in expression of genes that regulate the activation state. Costimulatory and modulatory molecules are considered with regard to their ability to affect the sensitivity or outcome of the BCR signal. Finally, we discuss how BCR signal transduction and the results of BCR signaling may differ at distinct stages in B-cell development. PMID- 9202889 TI - The role of statistics in cytopathology. PMID- 9202888 TI - Radiotherapy for palliation of symptoms in incurable cancer. AB - Approximately one half of prescribed radiotherapy is given for palliation of symptoms due to incurable cancer. Distressing symptoms including pain, bleeding, and obstruction can often be relieved with minimal toxic effects. Painful osseous metastasis is common in oncologic practice. Ninety percent of patients with symptomatic bone metastases obtain some pain relief with a lowdose, brief course of palliative radiotherapy. One half of the responding patients may experience complete pain relief. A single dose of 800 cGy in the setting of painful bone metastasis may provide pain control comparable to more protracted treatment at a higher dose of radiation. Patients with lytic disease in weight-bearing bones, particularly in the presence of cortical destruction, should be considered for prophylactic surgical stabilization of their condition. Routinely a brief, fractionated course of radiotherapy is given postoperatively. Pain due to multiple bone metastases uncontrolled by analgesics can be managed with single doses of halfbody irradiation. Doses of 600 cGy delivered to the upper half-body (above the umbilicus) and 800 cGy to the lower half-body (from the umbilicus to the middle of the femur) will provide some pain relief in 73% of patients. Half body techniques have been investigated as prophylactic treatment, as a complement to local-field irradiation, and as fractionated rather than singledose therapy. Although intravenous administration of strontium 89 has been associated with myelosuppression, this treatment has been shown (a) to relieve pain due to bone metastasis and (b) to delay development of new painful sites. Recent data from phase III trials demonstrated that bisphosphonates have a role in reducing skeletal morbidity due to bone metastasis. Bone pain was reduced, and the incidence of pathologic fracture and the need for future radiotherapy was decreased. Radiotherapy relieves clinical symptoms in 70% to 90% of patients with brain metastases. Brief treatment schedules (e.g., 2000 cGy in five fractions over 1 week) are as effective as more prolonged therapy. Patients with solitary brain metastasis and no extracranial disease or controlled extracranial disease should be considered for surgical resection, because phase III data indicate enhanced survival with such an approach. Whole-brain radiotherapy is routinely administered postoperatively. A phase III study is examining the impact of accelerated fractionated doses of radiotherapy (two treatments per day) on survival of patients with brain metastases. Stereotaxic radiosurgical treatment is becoming increasingly available and permits delivery of radiation to metastatic intracranial tumor with minimal exposure of normal surrounding brain This treatment is most commonly used at the time of a solitary recurrence of disease in patients who previously received whole-brain radiotherapy. A role for this modality in newly diagnosed brain metastases remains to be defined. Chest symptoms are common in patients with locally advanced lung cancer and are effectively palliated with one 1000 cGy or two 850 cGy one fraction doses of radiation to the thoracic inlet and mediastinum. Chest pain and hemoptysis are more effectively palliated than cough and dyspnea. In patients with stage III cancer there is no compelling evidence that radiotherapy confers a survival advantage, and it may be reasonable to administer thoracic radiotherapy only when the patient has significant symptoms and the goal is to achieve control of these symptoms. Approximately 75% of the cases of superior vena cava syndrome are due to lung cancer, and small-cell lung cancer is the most common histologic type. A histologic diagnosis should be obtained before treatment is started, because detection of lymphoma or small-cell carcinoma would necessitate systemic therapy. Eighty percent of the patients with vena cava syndrome due to malignant disease achieve symptom relief with a brief, fractionated, palliative course of rad PMID- 9202890 TI - In situ amplification in cytological preparations. PMID- 9202891 TI - A pilot study of cervical screening in an inner city area--lessons for a national programme. AB - The objectives of this study were to examine aspects of organization of a proposed national screening programme based in general practice. The target population of women aged 25-59 years and their general practitioners (GPs), in a defined inner city area, was identified from a population register of persons eligible for free medical services; a computerized system was developed for invitations and record linkage of cytology results. Smears were examined in one laboratory and follow up of women with abnormal smears was undertaken by one gynaecologist. A random sample of non-responders was surveyed by questionnaire. Response following two invitations was only 20%. Practices with male doctors only had significantly lower response rates (P < 0.001) than those with a female doctor/nurse. A survey of non-responders showed that over 20% of addresses were incorrect and 16% of those interviewed were ineligible for smear tests. A preference for a female to undertake smears was expressed by 67%, and 77% believed that the purpose of the cervical smear was to detect cancer. An accurate population register, health promotion, support for GP practices, provision of alternative venues for smear tests, development of computer systems, accurate data entry and fail-safe follow up are aspects of a cervical screening service which must be addressed prior to setting up a national service. PMID- 9202892 TI - DNA ploidy and pS2 protein expression in breast cancer. AB - The DNA content of ductal breast carcinomas of varying histological grade was measured using static image cytometry and correlated with pS2 expression in the tumour cells. Our study was performed on imprint of surgical biopsies of 60 women with ductal breast cancer. A statistically significant difference was observed between pS2+ expression and grade of malignancy (P < 0.001). The percentage of euploid tumours significantly decreased from grade I to grade II to grade III (P = 0.01). The percentage of aneuploid tumours increased from pS2+ to pS2- breast tumours (P < 0.001). These findings may be indicative of pS2 and DNA ploidy alterations and tumour aggressiveness. PMID- 9202893 TI - Phyllodes tumour: cytologic and histologic presentation of 22 cases, and immunohistochemical demonstration of p53. AB - A retrospective study of 22 cases of phyllodes tumour (PT) was undertaken to evaluate the potential value of fine needle aspiration (FNA) cytology in the diagnosis of benign and borderline PT. Histological material was available from 12 patients with typical benign PT (group 1), six patients with less typical changes (group 2) and four cases of borderline PT (group 3). Cytological presentation of PT in these cases was similar to that described by other cytologists, although abundant cellular material was obtained in only eight FNAs, naked nuclei were present in nine cases only, and atypical or suspicious cytological features were found in seven cases. Comparative analysis of p53 was made in nine patients with PT, five cases with other benign breast lesions and five with malignant lesions. p53 reaction was positive in five of nine patients with PT (all cases from groups 2 and 3), compared with two of five cases of carcinoma. p53 was negative in all patients with PT from group 1 and the five other benign cases. We suggest that cytopathologists should be careful when a myxoid stromal component is present in cytological smears. PMID- 9202894 TI - The effects of different sampling techniques on smear quality and the diagnosis of cytological abnormalities in cervical screening. AB - A major cause of false-negative cervical smear is sampling error. We examined the results obtained with three different instruments in 126,608 smears from general practitioners. The spatula/brush combination yielded the highest proportion of smears showing cytological abnormalities, and the Cervex brush the lowest. Although not a randomized study, this paper highlights the shortcomings of the Cervex brush. We postulate a mechanical deficiency. Diagnostic accuracy rather than a high proportion of good quality smears should dictate the choice of instrument. PMID- 9202895 TI - Angiomyolipoma of the liver: an unusual benign tumour identifiable on cytological material. PMID- 9202896 TI - Salivary epithelial-myoepithelial carcinoma: report of a case misinterpreted as pleomorphic adenoma on fine needle aspiration (FNA). PMID- 9202897 TI - Achievable standards, benchmarks for reporting and criteria for evaluating cervical cytopathology. PMID- 9202898 TI - The economics of screening. PMID- 9202899 TI - Charcot--Leyden crystals in fine needle aspiration (FNA) cytology of acute myeloid leukaemia. PMID- 9202900 TI - Rosette-like structures in a lymph node aspirate: pitfall in the diagnosis of neuroblastoma. PMID- 9202901 TI - Mesenchymal hamartoma of the chest wall. PMID- 9202902 TI - Management of abnormal glandular cytology on cervical smears. PMID- 9202903 TI - Importance of reporting adverse incidents relating to in vitro diagnostic medical devices. PMID- 9202904 TI - De Quervain's thyroiditis. PMID- 9202905 TI - Recommended code of practice for laboratories providing a cytopathology service 1997. British Society for Clinical Cytology. PMID- 9202906 TI - 'Off-label' use of prescription drugs: legal, clinical and policy considerations. PMID- 9202907 TI - Magnetic resonance imaging in children: role of the anaesthesiologist. PMID- 9202908 TI - Unilateral high frequency jet ventilation during one-lung ventilation. AB - Fifteen patients undergoing elective thoracic surgery were studied in order to investigate the efficacy of high frequency jet ventilation of the non-dependent lung with respect to arterial oxygenation. During the study PaO2, PaCO2, arterial pressures and heart rate were recorded during ventilation of both lungs in the lateral decubitus position during one-lung ventilation and during high frequency jet ventilation of the non-dependent lung. Mean PaO2 was 28 +/- 8.75 kPa and mean PaCO2 was 5.4 +/- 0.7 kPa during control. During one-lung ventilation, PaO2 dropped to 10.8 +/- 2.57 kPa and PaCO2 rose to 6.3 +/- 0.9 kPa. With high frequency jet ventilation to the non-dependent lung, mean PaO2 increased to 25 +/ 6.75 kPa and PaCO2 decreased to 5.16 +/- 0.9 kPa respectively. Arterial pressures and heart rate remained stable during the study period. In conclusion high frequency jet ventilation of the non-dependent lung was effective in providing arterial normoxaemia and normocapnia during one-lung ventilation. PMID- 9202909 TI - Midazolam for premedication in children: nasal vs. rectal administration. AB - The authors compared the acceptance and efficacy of rectal and nasal administration of midazolam (MDZ) for premedication. Ninety-five ASA I and II paediatric patients (8 months to 12 years) scheduled for elective surgery were randomly allocated to two groups. Group R received 0.3 mg kg-1 of rectal midazolam (in 5 mL saline). Group N received 0.2 mg kg-1 of nasal midazolam (5 mg ml-1). Both groups were divided in two subgroups according to age (group RA (< or = 6 years, n = 33), group RB (> 6 years, n = 18), group NA (< or = 6 years, n = 28), group NB (> 6 years, n = 16)). At the time of premedication, tolerance to the administration was confirmed. Twenty min after rectal or 10 min after nasal administration the quality of sedation was recorded. The nasal midazolam, in commonly used dosages, induced a sedation similar to that following rectal administration with a shorter delay of onset. Nasal administration was more often painful than rectal administration. Swallowing (nasal midazolam) and concerns about modesty (rectal midazolam) were more frequent in older children. Because of its poor tolerance, nasal premedication should be reversed for cases where there is no alternative. Rectal premedication should be avoided in older children. PMID- 9202910 TI - Intravenous administration of tenoxicam 40 mg for post-operative analgesia: a double-blind, placebo-controlled multicentre study. AB - The analgesic efficacy of tenoxicam, a newer injectable non-steroidal anti inflammatory drug, for post-operative analgesia after abdominal or orthopaedic surgery in ASA Grade I/II patients is reported. Two hundred and fifty-six patients received a single dose of tenoxicam 40 mg intravenous (i.v.) at the end of surgery and this was repeated 24 h later. These patients were compared, with respect to pain or adverse events, with 258 patients that received placebo. All patients were monitored for the next 72 h. Overall, tenoxicam provided reliable analgesia with comparable pain scores at rest, moving and coughing. The cumulative rescue PCA-morphine consumption was always lower in the tenoxicam treated patients and was most marked at 4 and 24 h after the second injection of tenoxicam. This effect was more pronounced after abdominal surgery. The intravenous administration of tenoxicam was associated with a low incidence of adverse events and a high tolerability. PMID- 9202911 TI - The effect of general anaesthesia on breathing patterns in elderly patients during the early post-operative period. AB - This study was designed to investigate the incidence of critical events in breathing following light general anesthesia compared with normal sleep during the first 12 h after transfer from the recovery room. There were no significant differences in the incidence of apnoea or desaturation episodes between normal sleep and the post-operative recovery period. There was a close correlation between the pre-operative and post-operative incidence of apnoea (r = 0.93), pre operative and post-operative desaturation periods (r = 0.81), pre-operative and post-operative mean SpO2 values (r = 0.54) and pre-operative and post-operative minimal SpO2 values (r = 0.90) in all the patients. In the early post-operative period, breathing patterns and oxygenation were similar to those observed during normal night-time sleep in elderly patients undergoing ophthalmological surgery. PMID- 9202912 TI - Haemodynamic effects of pneumoperitoneum in elderly patients with an increased cardiac risk. AB - We studied the haemodynamic changes induced by pneumoperitoneum (PP) in elderly patients with increased cardiac risk (ASA class III; n = 10; age 72.3 +/- 8.8 years, mean +/- SD, P < 0.05; group 2) and compared the results with patients at normal risk (ASA class I, II; n = 12; age 55.6 +/- 11.8 years; group 1). Thermodilution measurements were performed after induction of general anaesthesia (T1), after onset of PP (T2, intraabdominal pressure 14 mmHg) and after additional 15 degrees head-up tilt (T3). In both groups PP, as compared with T1, induced a significant increase in mean arterial pressure (MAP, mmHg, group 1: 77 +/- 14 to 96 +/- 18, P < 0.05/group 2: 75 +/- 10 to 102 +/- 18, P < 0.01), mean pulmonary artery pressure (MPAP, mmHg: 15 +/- 5 to 22 +/- 4, P < 0.01/18 +/- 3 to 25 +/- 5, P < 0.01), central venous pressure (CVP, mmHg: 7 +/- 2 to 15 +/- 3, P < 0.01/7 +/- 2 to 12 +/- 2, P < 0.01), pulmonary capillary wedge pressure (PCWP, mmHg: 9 +/- 4 to 16.3, P < 0.01/8 +/- 2 to 15 +/- 6, P < 0.01) and in systemic vascular resistance (SVR, dynes s cm-5: 1415 +/- 375 to 1873 +/- 412, P < 0.01/ 1502 +/- 360 to 2067 +/- 647, P < 0.01). Cardiac index (CI, L min-1 m-2: 2.3 +/- 0.3 to 1.9 +/- 0.3, P < 0.05/2.2 +/- 0.4 to 2.2 +/- 0.5 P = 0.76) and oxygen delivery index (DO2I, mL min-1 m-2: 388 +/- 54 to 324 +/- 61, P < 0.05/358 +/- 69 to 353 +/- 82, P = 0.77) decreased in group 1 but not in group 2. Heart rate, stroke Index, pulmonary vascular resistance, arteriovenous oxygen content difference and oxygen consumption index were unchanged. After head-up tilt MAP (mmHg, 92 +/- 15, P < 0.05/ 101 +/- 17, P < 0.01), MPAP (mmHg, 20 +/- 3, P < 0.01/22 +/- 4, P < 0.05), CVP (mmHg, 12 +/- 2, P < 0.01/10 +/- 2, P < 0.01) and PCWP (mmHg, 12 +/- 3, P < 0.05/12 +/- 5, P < 0.05) remained elevated compared with T1 in both groups, SVR (dynes s cm-5, 1575 +/- 372, P = 0.13/1793 +/- 528, P < 0.01) in group 2 only. No complications occurred. The results indicate that PP is associated with significant but relatively benign haemodynamic changes. Anaesthesia for laparoscopic cholecystectomy may be performed safely also in elderly ASA class III patients with increased cardiac risk. An adequate haemodynamic monitoring is recommended. PMID- 9202913 TI - A survey of factors determining the utilization of autologous blood donation in hip replacement surgery. AB - A prospective survey was conducted among anaesthesiologists to determine their criteria for excluding a patient from autologous blood donation prior to total hip prosthesis. Information on patients operated on during a 5-week period was obtained using a questionnaire, consisting of a set of possible responses to simple questions, matched with some parametric values. Sixty-eight of the 99 patients included in the survey underwent surgery without donation. Age was the main cause for exclusion (34%). The age of patients excluded because of their age alone was 75 +/- 11 years (standard deviation). The age of patients excluded from donation and who did not subsequently receive homologous blood was comparable with that of patients not autotransfused but who received homologous blood (68 +/ 4 vs. 69 +/- 6 years, respectively). Post-operative haemoglobin concentrations were similar whether transfusion was performed or not, and regardless of the origin of blood given. Since age is no longer taken into consideration at the time of transfusion, it is likely that exclusion of a patient from autologous blood donation because of age is an arbitrary choice. PMID- 9202914 TI - The optimal dose of local anaesthetic in the orthogonal two-needle technique. Extent of sensory block after the injection of 20, 30 and 40 mL of anaesthetic solution. AB - Ninety patients undergoing scheduled upper limb orthopaedic surgery were studied to determine the optimal anaesthetic dose using the 'orthogonal two-needle technique'. The patients were randomly assigned to one of three groups to receive one of three different volumes (20, 30 and 40 mL) (n = 30) of anaesthetic solution (a mixture of equal parts of 0.5% bupivacaine with adrenaline 1:200,000 and 2% lignocaine). A significant correlation was found between the volume injected and the anaesthetic spread for all tested areas. A better analgesic spread to all the major branches of the plexus was obtained when increased volumes of anaesthetic solution were injected. The comparisons between the 20 mL group and the other two groups are significant in all the tested areas, as well as the comparisons between 30 and 40 mL groups in the areas innervated by radial and musculocutaneous nerves. Only the area innervated by the axillary nerve showed a weaker volume-analgesia relation, confirming the elusiveness of this area to anaesthesia in the axillary approaches. The improved results observed using greater amounts of anaesthetic solution might result from a higher intrasheath pressure with disruption of sheath septa, or from a greater availability of drug for all the terminal branches of brachial plexus, or both. PMID- 9202915 TI - Patients' refusal to participate in clinical research. AB - The number of protocol-eligible patients, refusing to participate in a clinical trial is often not mentioned. The aim of this study is to report the number of refusers and to evaluate the reasons for not participating in a clinical study concerning post-operative pain relief and to assess the potential influence on the final study results. Patients refusing to participate in the study were recorded and evaluated for reasons of refusal. The post-operative pain relief techniques applied in this trial are commonly used, but nevertheless the refusal rate was higher than expected. When it was mentioned that an epidural technique was a part of the trial, 16.7% of the total protocol-eligible group refused. The responses of those offered an epidural could be divided into two groups: the adamant pros and cons to this technique. It can be concluded that in order to be able to judge the validity of results and thus for good clinical practice, the number of patients refusing to participate in a clinical trial and their reasons, should be mentioned in all publications. PMID- 9202916 TI - Double-blind comparison of alizapride, droperidol and ondansetron in the treatment of post-operative nausea. AB - To compare the efficacy in the treatment of post-operative nausea and/or vomiting (PONV), 75 patients undergoing gynaecological procedures under general anaesthesia using N2O/enflurane who suffered from PONV in the first hour after surgery were randomly allocated to three groups containing 25 patients each to receive either alizapride 100 mg, droperidol 1 mg or ondansetron 8 mg (i.v.). Patients expressed the severity of their nausea on a Visual Analogue Scale (VAS) ranging from 0 (none) to 10 (as bad as possible). Vomiting was recorded as present or absent, and the number of emetic events was noted. Data were recorded until rescue medication was given or until 4 h after the administration of the study drug. There were no significant differences between the three groups in the average VAS scores and the presence of vomiting at the time of entry into the study. Fifteen and 30 min after the administration of the study drug, VAS decreased notably in all groups. This decreases was similar and statistically significant within each group. However, comparison between the three groups showed no statistically significant differences. There was no statistically significant difference between the three groups in the number of patients receiving rescue medication, the number of emetic events and the time from administration of the study drug until rescue medication was given. It is concluded that alizapride 100 mg, droperidol 1 mg and ondansetron 8 mg intravenously are equally effective in the treatment of PONV after gynaecological procedures and that the newer drugs alizapride and ondansetron offer no advantage over droperidol. PMID- 9202917 TI - Nitrous oxide inhalation as an adjunct to intravenous induction of general anaesthesia with propofol for day surgery. AB - Fifty patients were randomly allocated to receive either a preinduction inhalation with nitrous oxide (50%) in oxygen or fentanyl with preoxygenation, before induction of anaesthesia with propofol. Both groups of patients showed a significant rise in arterial oxygen saturation prior to propofol induction which established similar depths of anaesthesia, determined by the acceptability of the laryngeal mask placement. In the fentanyl group there was a significant period of apnoea after induction, with 40% of the patients being apnoeic for 120s or more and requiring assisted ventilation. Reduction in arterial blood pressure was also more rapid in the fentanyl group compared with the nitrous oxide group. Preinduction inhalation of nitrous oxide (50%) in oxygen appears to be an effective and acceptable method of preoxygenating the patient and augmenting the propofol induction of anaesthesia. PMID- 9202918 TI - Haemodynamic monitoring during alkalinized lignocaine epidural block: a comparison with subarachnoid anaesthesia. AB - Cardiovascular responses after epidural alkalinized lignocaine and subarachnoid hyperbaric bupivacaine administration were studied using a non-invasive cardiac output measurement in 32 ASA Grade I-II patients undergoing orthopaedic leg surgery (hip hemi-arthroplasty or Ender nailing). All patients achieved adequate surgical anaesthesia. The block onset time was faster (P = 0.003), and the range of final sensory level wider (P = 0.006) in patients receiving spinal anaesthesia compared with the epidural group. Diastolic arterial pressure was significantly reduced when compared with base-line (P = 0.002) only in the spinal group. No significant changes in stroke volume, systemic vascular resistance or left ventricular stroke work were observed in either group. Heart rate and cardiac index were significantly reduced in the spinal group when compared both with base line (P = 0.002; P = 0.04) and the epidural group (P = 0.001; P = 0.006). The results demonstrated that the block onset time and the cardiovascular effects produced by lumbar epidural anaesthesia, with alkalinized solutions, remain less than after spinal anaesthesia involving the same segments. PMID- 9202919 TI - Bradycardia following intravenous ketorolac in children. AB - Two cases of bradycardia following the intra-operative use of ketorolac given intravenously (i.v.) to children are reported. We examine why this may not have been previously reported and make recommendations regarding its use. PMID- 9202920 TI - The effect of propofol and thiopentone on impairment by reactive oxygen species of endothelium-dependent relaxation in rat aortic rings. AB - The effect in isolated rat aorta of propofol, thiopentone, midazolam, etomidate and fentanyl on the impairment of endothelium-dependent relaxation by reactive oxygen species is reported. Aortic rings were exposed to reactive oxygen species by the electrolysis of the bathing physiological buffer, and endothelium dependent relaxation in response to acetylcholine was inhibited. This inhibition was countered by incubation before electrolysis with propofol (2 x 10(-5) and 6 x 10(-5) M) or thiopentone (10(-5)-10(-4) M), but not with midazolam (3 x 10(-4) M), etomidate (3 x 10(-4) M) or fentanyl (3 x 10(-5) M). We suggest that the protective effect of propofol and thiopentone against the loss of endothelium dependent relaxation caused by reactive oxygen species may be because of their antioxidant and free radical scavenging properties, which could be clinically relevant during surgical procedures in which ischaemia is unavoidable. PMID- 9202921 TI - Clonidine inhibits gastric motility in the rat. AB - We have studied the effect of clonidine on gastric motility, by examining the effect on gastric emptying of indigestible solids. In Wistar rats, either clonidine or saline was injected intraperitoneally, and ten steel balls (1.0 mm in diameter) were inserted into the stomach. Gastric emptying was examined at 3 h. Clonidine delayed gastric emptying of steel balls (ED50 = 0.0071 [95% confidence interval: 0.0033-0.013) mg kg-1). Yohimbine, but not naloxone, significantly antagonized the inhibitory effect of clonidine. We conclude that clonidine inhibits gastric motility through the alpha 2 adrenoceptor. PMID- 9202922 TI - Propofol neuroprotection in a rat model of ischaemia reperfusion injury. AB - This study was designed to test the hypothesis that propofol, which possesses antioxidant properties, would produce greater protection than isoflurane in cerebral ischaemia reperfusion injury, at dose levels that produced similar affects on brain electrical activity. Twenty Sprague-Dawley rats were anaesthetized using isoflurane 1.5% in air/oxygen, and mechanically ventilated to a PaCO2 of 4.5 kPa. Following 90 min of middle cerebral artery occlusion using an intraluminal filament, rats were given, in random order, either propofol 1 mg kg 1 min-1 or isoflurane 3% (both of which have been shown to reliably produce EEG burst suppression). After a further 30 min, reperfusion was induced by withdrawing the filament. Animals were killed following 240 min of reperfusion. Rats in the propofol group showed a 21% reduction in mean hemispheric infarct volume when compared with the isoflurane group (P < 0.0001). These data suggest that propofol may act by mechanisms in addition to CMRO2 depression, and provide a basis for further studies of propofol neuroprotection. PMID- 9202923 TI - Visual evaluation of train-of-four and double burst stimulation, fade at various currents, using a rubber band. AB - The sensitivity of train-of-four (TOF) or double burst stimulation3,3 (DBS3,3) was examined to detect fade by visual inspection, at varying stimulating currents, using the thumb of the investigated arm maintained abducted by the use of a rubber band. One-hundred adult patients were allocated randomly to (1) train of-four-rubber band (TOF-RB), (2) train-of-four-control (TOF-control), (3) double burst stimulation-rubber band (DBS-RB), or (4) double burst stimulation-control (DBS-control) group. Each group contained 25 patients. The thumb of the investigated arm was kept abducted using a rubber band in the TOF-RB band and DBS RB group and was free in the TOF-control and DBS-control. TOF stimuli were delivered in the TOF-RB and TOF-control groups, and DBS3,3 in the TOF-RB and TOF control at 50, 30 and 20 mA. The probability of visual detection of TOF fade was the same in the TOF-RB and TOF-control groups. In contrast, when measured TOF ratio was 0.51-0.80, at the stimulating current of 50 or 30 mA, the likelihood of visual detection of fade in the DBS-RB group was significantly higher than in the DBS-control group (P < 0.05). With a TOF ratio of 0.61-0.70, in the DBS-RB group the probability of visual detection of fade at 50 or 30 mA was significantly higher than at 20 mA (P < 0.05). This study suggests that when using a rubber band, fade in response to the DBS3,3 is detected by visual inspection more readily at a stimulating current of 50 or 30 mA than without the rubber band. PMID- 9202924 TI - Factors affecting flow through blood administration sets. AB - Factors affecting flow through blood administration sets in vitro were assessed under gravity-fed and pressurized conditions including an assessment of the influence of the intravenous (i.v.) cannula and Luer lock fitting. The fastest gravity fed flow of 4.775 mL s-1 was obtained through the largest internal diameter (ID = 4.8 mm) blood administration set. Flow through blood administration sets with ID = 3 mm was approximately 50% of this. Flow increased over base-line through all the administration sets when the i.v. cannula was removed (range 18-50%) and increased further over base-line when the Luer lock fitting was removed from the distal end (range 26-129%), indicating that these are rate-limiting steps in the system. The Y-type trauma set with the largest diameter tubing facilitated the fastest flow, although flow through all the Y type trauma sets produced lower flow rates than the corresponding blood administration sets, which may reflect their relative increased length. The ideal blood administration set should have an internal diameter at least 4 mm and be less than 170 cm in length. PMID- 9202925 TI - Acute laryngeal obstruction--reaction to intravenous hydrocortisone? PMID- 9202926 TI - Transient blindness after TURP. PMID- 9202929 TI - Anaesthesiological manpower in Europe. PMID- 9202927 TI - Safe and simple, an alternative technique for anaesthesia during magnetic resonance imaging (MRI) PMID- 9202930 TI - Update in intravenous anaesthesia. Proceedings of the 7th International Symposium on Intravenous Anaesthesia. Lausanne, Switzerland, 2-3 May 1997. PMID- 9202932 TI - Mechanisms of actions of opioids and non-steroidal anti-inflammatory drugs. AB - Opioids and non-steroidal anti-inflammatory drugs (NSAIDs) are the commonest drugs used to treat pain. Opioids mimic the actions of endogenous opioid peptides by interacting with mu, delta or kappa opioid receptors. The opioid receptors are coupled to G1 proteins and the actions of the opioids are mainly inhibitory. They close N-type voltage-operated calcium channels and open calcium-dependent inwardly-rectifying potassium channels. This results in hyperpolarization and a reduction in neuronal excitability. They also decrease intracellular cAMP which modulates the release of nociceptive neurotransmitters (e.g. substance P). Inhibition of prostaglandin synthesis by cyclooxygenase is the principal mode of the analgesic and anti-inflammatory actions of NSAIDs. Cyclo-oxygenase is inhibited irreversibly by aspirin and reversibly by other NSAIDs. The widespread inhibition of cyclo-oxygenase is responsible for many of the adverse effects of these drugs. NSAIDs also reduce prostaglandin production within the CNS. This is the main action of paracetamol. PMID- 9202931 TI - Intravenous anaesthetics: some cellular sites of action. AB - Intravenous anaesthetics have diverse effects on neurones within the central nervous system. Only those that occur at clinical concentrations are likely to be relevant. The dominant effect of many agents is the potentiation of the inhibitory neurotransmitter gamma amino butyric acid (GABA) by various mechanisms while inhibiting the effects of excitatory transmitters seems to be less dominant, except for ketamine. PMID- 9202933 TI - Propofol: pro- or anticonvulsant? AB - The pro- or anticonvulsant properties of propofol remain a matter of controversy. Although numerous case reports describe the appearance of abnormal movements, posturing and seizure-like activity related to the use of propofol, systematic studies in both humans and animals strongly suggest that it possesses antiepileptic properties. Propofol consistently reduces the seizure duration during electroconvulsive therapy, its use has been successful in controlling refractory status epilepticus and in animals it offers a strong protection against lignocaine- or pentylene-tetrazol-induced epilepsy. The beneficial effects of propofol may be related to its uniform depressant action on the central nervous system, to a potentialization of GABA-mediated pre- and postsynaptic inhibition, and by decreasing the release of excitatory transmitters, glutamate and aspartate. PMID- 9202934 TI - Monitoring depth of anaesthesia. AB - In clinical practice, indirect and non-specific signs are used for monitoring anaesthetic adequacy. These include haemodynamic, respiratory, muscular and autonomic signs. These measures do not indicate adequacy of anaesthesia in a reliable manner. Many attempts have been made to find a more accurate monitor. Direct monitoring of anaesthetic effect should be possible by EEG measurement. EEG information can be reduced, condensed and simplified, leading to single numbers (spectral edge frequency and median frequency). These methods appear insufficient for assessing anaesthetic adequacy. The bispectral index, derived from bispectral analysis of the EEG, is a very promising tool for measuring adequacy of anaesthesia. An alternative approach is to monitor evoked potentials. Middle latency auditory evoked potentials may be helpful in assessing anaesthetic adequacy. Both techniques need further validation. PMID- 9202935 TI - Target-controlled anaesthesia: concepts and first clinical experiences. AB - Propofol has a favourable pharmacokinetic profile for total intravenous anaesthesia and several manual infusion schemes have been proposed to maintain a constant blood concentration during anaesthesia. However, such schemes cannot respond predictably to changing surgical and anaesthetic requirements. A pharmacokinetic model for propofol has been incorporated into a target-controlled infusion system. This allows the desired target blood concentration appropriate for any individual patient and level of surgical stimulation to be achieved and maintained at any time. There is no single blood concentration of an anaesthetic agent which will result in satisfactory anaesthesia for all patients and all surgical conditions. It is necessary to titrate the target concentration against each patient's clinical response. Target-controlled systems provide the best estimate of the blood concentration at any time and permit the required target concentration to be achieved as accurately and as rapidly as possible. PMID- 9202936 TI - How to manage drug interactions. AB - Multiple drugs are used to provide anaesthesia. On average, four to six drugs are used during anaesthesia and, therefore, drug interactions are common. These interactions are primarily either pharmacokinetic or pharmacodynamic. Due to the relatively short duration of drug administration for anaesthesia, pharmacokinetic drug interactions resulting from alterations in drug metabolism do not generally produce clinically significant effects. Pharmacodynamic-drug interactions between anaesthetic drugs, however, are potentially serious. This may reflect that anaesthesia is not a single entity, but a process provided by a combination of drugs; i.e. loss of consciousness, analgesia and neuromuscular blockade. An understanding of each drug's pharmacokinetics, pharmacodynamics and drug interactions will allow clinicians to administer drugs to provide a more optimal anaesthetic. PMID- 9202937 TI - Remifentanil: when and how to use it. AB - Remifentanil is a new potent mu-agonist with a unique pharmacokinetic profile due to a rapid metabolism by non-specific tissue esterases. As a consequence, remifentanil pharmacokinetics are not modified by severe renal or hepatic dysfunction. During general anaesthesia, any dosage of remifentanil may be used without undue lengthening of emergence times. In cardiac surgery, remifentanil combines the requirement for intra-operative control of stress responses and rapid recovery. The rapid termination of remifentanil action warrants modifications of the current practice concerning early postoperative pain control. Remifentanil may be used as a sedative during monitored analgesia, or as a postoperative analgesic in spontaneously breathing patients, provided bolus doses are avoided. Remifentanil may increase patients' safety by eliminating the risk of delayed respiratory depression, but its correct use requires major changes in our prescribing habits. PMID- 9202938 TI - The psychological and psychiatric consequences of the ICU stay. AB - From the patient's point of view, the Intensive Care Unit is both a frightening place and a safe haven. Psychologically, what we see most commonly are regression, delirium and paranola. Regression requires no treatment; delirium is treatable not only medically but psychologically as well; paranoia is best treated by prevention. PMID- 9202939 TI - Pain treatment in the ICU: intravenous, regional or both? AB - Adequate treatment of pain in ICU patients should be an integral part of ICU management, as inadequately treated pain leads to a series of complications that may counteract the success of ICU treatment. For continuous intravenous use we recommend sufentanil in a dose of 0.75-1.0 microgram kg-1 h-1 in mechanically ventilated patients and in a dose of 0.25-0.35 microgram kg-1 h-1 in intubated and spontaneously breathing patients. On-demand analgesia, administered via the intravenous or epidural route, may be an alternative to a relatively fixed continuous infusion of an analgesic drug, and in some ICU patients the transdermal use of opioids can be an alternative to continuous intravenous drug application or PCA. Increased sizes of the patch (25, 50, 75, 100 cm2) provide sustained transdermal rates of approximately 25, 50, 75 and 100 micrograms h-1 of fentanyl over a period up to about 72 hours. Patients with trauma to the thorax, pelvic fracture, or after major surgical interventions will be better managed by regional application of analgesic drugs alone or in combination with a systemic analgesic drug infusion. To achieve the best results it is necessary to be well informed and trained in the method, to know the advantages and disadvantages, the correct and modified dosages of the drugs used, and the indications and contraindications. PMID- 9202940 TI - Neuromuscular blockade: is it still useful in the ICU? AB - Neuromuscular relaxants may be administered to ICU patients for days or weeks. The most frequent indications are facilitation of mechanical ventilation and management of neurosurgical patients after severe head injury. Tachyphylaxis can be observed in ICU patients but the major problem remains the prolonged weakness which can occur after long-term administration of muscle relaxants. The mechanisms are still poorly understood and under investigation. Although neuromuscular monitoring provides guidelines for the administration of muscle relaxants in critically ill patients, it does not prevent the development of a myopathy. PMID- 9202941 TI - Cold induced vasodilatation and cardiovascular responses in humans during cold water immersion of various upper limb areas. AB - To study the physiological responses induced by immersing in cold water various areas of the upper limb, 20 subjects immersed either the index finger (T1), hand (T2) or forearm and hand (T3) for 30 min in 5 degrees C water followed by a 15 min recovery period. Skin temperature of the index finger, skin blood flow (Qsk) measured by laser Doppler flowmetry, as well as heart rate (HR) and mean arterial blood pressure (BPa) were all monitored during the test. Cutaneous vascular conductance (CVC) was calculated as Qak/BPa. Cold induced vasodilatation (CIVD) indices were calculated from index finger skin temperature and CVC time courses. The results showed that no differences in temperature, CVC or cardiovascular changes were observed between T2 and T3. During T1, CIVD appeared earlier compared to T2 and T3 [5.90 (SEM 0.32) min in T1 vs 7.95 (SEM 0.86) min in T2 and 9.26 (SEM 0.78) min in T3, P < 0.01]. The HR was unchanged in T1 whereas it increased significantly at the beginning of T2 and T3 [+13 (SEM 2) beats.min-1 in T2 and +15 (SEM 3) beats.min-1 in T3, P < 0.01] and then decreased at the end of the immersion [-12 (SEM 3) beats.min-1 in T2, and -15 (SEM 3) beats.min-1 in T3, P < 0.01]. Moreover, BPa increased at the beginning of T1 but was lower than in T2 and T3 [+9.3 (SEM 2.5) mmHg in T1, P < 0.05; +20.6 (SEM 2.6) mmHg and 26.5 (SEM 2.8) mmHg in T2 and T3, respectively, P < 0.01]. The rewarming during recovery was faster and higher in T1 compared to T2 and T3. These results showed that general and local physiological responses observed during an upper limb cold water test differed according to the area immersed. Index finger cooling led to earlier and faster CIVD without significant cardiovascular changes, whereas hand or forearm immersion led to a delayed and slower CIVD with a bradycardia at the end of the test. PMID- 9202942 TI - Assessment of cardiorespiratory exercise function in obese children and adolescents by body mass-independent parameters. AB - The parameters used to assess aerobic exercise function by gas exchange are usually adjusted for body mass and are expressed as millilitres per minute per kilogram. In the case of obese children this could lead to overcorrection with an underestimation of their exercise capacity. The purpose of the present study was to assess cardiorespiratory exercise function in obese subjects using body mass independent parameters. As both carbon dioxide output (VCO2) and oxygen uptake (VO2) are usually corrected for body mass, the slope of VCO2 versus VO2 can be considered to be independent of body mass. This slope was calculated below the ventilatory threshold (S1) and above the ventilatory threshold (S3). Exercise tests were performed on a treadmill and respiratory gas exchange was measured breath-by-breath. A group of 29 obese children [mean age 11 (SD 2.5) years] were compared to 16 normal controls of the same age range [mean age 10.8 (SD 2.2); P > 0.05]. The patients were overweight by 36 (SD 17.9)% and had a body mass index of 25.0 (SD 3.8). The results showed that S3 in the obese subjects was significantly steeper compared to the normal controls [1.30 (SD 0.20) vs 1.10 (SD 0.20); P < 0.05]. The steepest values for S3 were found in the subjects with the highest degree of obesity. This method has some limitations, since in a large proportion of the patients (48%) no ventilatory threshold could be detected, which is prerequisite for calculation of these slopes. The latter was already suppressed at the onset of exercise in 21% of the sample or could not be detected because of breathing irregularity in 27%. It is suggested from this study that cardiorespiratory exercise function in obese children is reduced, especially when assessed by parameters of aerobic exercise which cancel the confounding effect of body mass. PMID- 9202943 TI - Muscle performance and electromyogram activity of the lower leg muscles with different levels of cold exposure. AB - The purpose of this study was to evaluate the relationship between different levels of body cooling and muscle performance decrement and to study the motor co ordination of the working agonist-antagonist muscle pair of the lower leg. Eight volunteer male subjects dropped from a 40-cm bench on to a force plate and performed a maximal rebound jump (stretch-shortening cycle). The jumps were performed after 60-min exposures to 27 degrees C, 20 degrees C, 15 degrees C and 10 degrees C. In comparison to those at 27 degrees C, all the exposures to lower temperatures decreased the flight time of the jump, average force production and take-off velocity in a dose-dependent manner. The changes in electromyogram (EMG) activity also behaved in a dose-dependent manner. During pre-activity and stretch phases the integrated EMG (iEMG) activity of the agonist muscle (triceps surae) increased due to cooling (at 10 degrees C, P < 0.05). In contrast, during the shortening phase iEMG of the agonist muscle decreased due to cooling (at 15 degrees C and 10 degrees C, P < 0.05). Moreover, the activity of the antagonist muscle (tibialis anterior) increased due to cooling (at 15 degrees C and 10 degrees C, P < 0.01). The mean power frequency of the agonist muscle during the shortening phase was shifted from 124 (SEM 12) Hz (at 27 degrees C) to 82 (SEM 7) Hz (at 10 degrees C, P < 0.01). We concluded that there was a dose-dependent response between the degree of cooling and the amount of decrease in muscle performance as well as EMG activity changes. A relatively low level of cooling was sufficient to decrease muscle performance significantly. PMID- 9202944 TI - Performance and fibre characteristics of human skeletal muscle during short sprint training and detraining on a cycle ergometer. AB - The ergometric effect of sprint training and detraining was studied in relation to muscle fibre changes in seven students trained during 9 weeks on a cycle ergometer. Before and after training and after 7-week detraining, they performed a force-velocity test on a friction-loaded cycle ergometer. On these three occasions, muscle samples were taken from vastus lateralis muscle at rest for histochemical analysis. The training-induced shift of the force-velocity relationship was such that the increase in maximal velocity (vmax) was greatest against high braking forces (FB) with unchanged vmax with no load. This was associated with higher maximal power output (28%) and peak force (16%). The increased maximal mean power output to reach a maximal velocity during a short sprint was obtained against a 23% higher optimal FB (FB,Wmax). At the same time, a considerable hypertrophy in fast twitch b (FTb) fibres was observed. All these changes were maintained after detraining. The training-induced changes in vmax reached against FB1Wmax(vm2Wmax) allowed us to produce evidence for two particular sub-groups in which inverse fibre conversions were observed. In subgroup A, the lowered post-training vm,Wmax was associated with a decrease in both FTa and FTb fibres. Conversely, the vm,Wmax, increase in subgroup B was associated with a higher percentage of FT fibres as the result of increased FTa fibres and decreased FTb fibres. Thus, the fibre hypertrophy associated with a unidirectional fibre translation [FTb-->FTa-->slow twitch (ST)] toward fibres with a high thermodynamic efficiency would result mainly in increased force qualities, whereas the bidirectional fibre translation (ST-->FTa<--FTb) would allow enhancement of both force and velocity properties. PMID- 9202945 TI - Effects of aerobic exercise on the torque-velocity relationship in cycling. AB - The kinetics of the torque-velocity (T-omega) relationship after aerobic exercise was studied to assess the effect of fatigue on the contractile properties of muscle. A group of 13 subjects exercised until fatigued on a cycle ergometer, at an intensity which corresponded to 60% of their maximal aerobic power for 50 min (MAP60%); ten subjects exercised until fatigued at 80% of their maximal aerobic power for 15 min (MAP80%). Of the subjects 7 exercised at both intensities with at least a 1-week interval between sessions. Pedalling rate was set at 60 rpm. The T-omega relationship was determined from the velocity data collected during all-out sprints against a 19 N.m braking torque on the same ergometer, according to a method proposed previously. Maximal theoretical velocity (omega zero) and maximal theoretical torque (Tzero) were estimated by extrapolation of the linear T-omega relationship. Maximal power (Pmax) was calculated from the values of Tzero and omega zero (Pmax = 0.25 omega zero Tzero). The T-omega relationships were determined before, immediately after and 5 and 10 min after the aerobic exercise. The kinetics of omega zero, Tzero and Pmax was assumed to express the effects of fatigue on the muscle contractile properties (maximal shortening velocity, maximal muscle strength and maximal power). Immediately after exercise at MAP60% a 7.8% decrease in Tzero and 8.8% decrease in Pmax was seen while the decrease in omega zero was nonsignificant, which suggested that Pmax decreased in the main because of a loss in maximal muscle strength. In contrast, MAP80% induced a 8.1% decrease in omega zero and 12.8% decrease in Pmax while the decrease in Tzero was nonsignificant, which suggested that the main cause of the decrease in Pmax was probably a slowing of maximal shortening velocity. The short recovery time of the T-omega relationship suggests that the causes of the decrease of torque and velocity are processes which recover rapidly. PMID- 9202946 TI - Analysis of the tonic vibration reflex: influence of vibration variables on motor unit synchronization and fatigue. AB - The influence of vibration frequency (40, 80, 100, 120, 150, or 200 Hz) at selected displacement amplitudes (0.2, 0.3 mm) on tonic vibration reflex (TVR) characteristics was investigated. The degree of synchronization of motor unit activity with vibratory stimuli in ten humans was determined using the electromyographic (EMG) activity of the finger and wrist flexor muscles when vibration was applied to the distal tendons of the hand flexor muscles. The EMG spectral analysis indicates that harmonic and subharmonic motor unit synchronization mechanisms contribute to the modulation of the amplitude of the TVR as the vibration frequency increases. Harmonic synchronization decreases while subharmonic synchronization increases as vibration frequency increases. It is suggested that the synchronization process influences muscle fatigue, since it forces the driving of motor units, leading to a decrease in contraction efficiency. This phenomenon most probably results from an impairment of excitation-contraction coupling. High-frequency vibration (> 150 Hz) tends to induce less motor unit synchronization in a frequency range beyond the known mechanical resonance of biological tissues. The findings of this study may be applied to the design of hand-held power tools, since their vibration triggers the TVR in active muscles. PMID- 9202947 TI - The influence of maximal aerobic power on recovery of skeletal muscle following anaerobic exercise. AB - Phosphorus magnetic resonance spectroscopy (31P-MRS) was used to investigate the influence of maximal aerobic power (VO2max) on the recovery of human calf muscle from high-intensity exercise. The (VO2max) of 21 males was measured during treadmill exercise and subjects were assigned to either a low-aerobic-power (LAP) group (n = 10) or a high-aerobic-power (HAP) group (n = 11). Mean (SE) VO2max of the groups were 46.6 (1.1) and 64.4 (1.4) ml.kg-1.min-1, respectively. A calf ergometry work capacity test was used to assign the same relative exercise intensity to each subject for the MRS protocol. At least 48 h later, subjects performed the rest (4 min), exercise (2 min) and recovery (10 min) protocol in a 1.5 T MRS scanner. The relative concentration of phosphocreatine (PCr) was measured throughout the protocol and intracellular pH (pHi) was determined from the chemical shift between inorganic phosphate (Pi) and PCr. End-exercise PCr levels were 27 (3.4) and 25 (3.5)% of resting levels for LAP and HAP respectively. Mean resting pHi was 7.07 for both groups, and following exercise it fell to 6.45 (0.04) for HAP and 6.38 (0.04) for LAP. Analysis of data using non-linear regression models showed no differences in the rate of either PCr or pHi recovery. The results suggest that VO2max is a poor predictor of metabolic recovery rate from high-intensity exercise. Differences in recovery rate observed between individuals with similar VO2max imply that other factors influence recovery. PMID- 9202948 TI - The effect of a thiamin derivative on exercise performance. AB - The purpose of this study was to investigate the effect of a thiamin derivative, thiamin tetrahydrofurfuryl disulfide (TTFD), on oxygen uptake (VO2), lactate accumulation and cycling performance during exercise to exhaustion. Using a randomized, double-blind, cross-over design with a 10-day washout between trials, 14 subjects ingested either 1 g.day-1 of TTFD or a placebo (PL) for 4 days. On day 3, subjects performed a progressive exercise-test to exhaustion on a cycle ergometer for the determination of VO2submax, VO2peak, lactate concentration ([La ]), lactate threshold (ThLa) and heart rate (fc). On day 4, subjects performed a maximal 2000-m time trial on a cycle ergometer. A one-way analysis of variance (ANOVA) with repeated measures was used to determine significant differences between trials. There were no significant differences detected between trials for serial measures of VO2submax, [La-] or fc. Likewise, VO2peak [PL 4.06 (0.19) TTFD 4.12 (0.19) l.min-1, P = 0.83], ThLa [PL 2.47 (0.17), TTFD 2.43 (0.16) l.min-1, P = 0.86] and 2000-m performance time [PL 204.5 (5.5), TTFD 200.9 (4.3).s, P = 0.61] were not significantly different between trials. The results of this study suggest that thiamin derivative supplementation does not influence high-intensity exercise performance. PMID- 9202950 TI - Reliability of an electrophoretic and image processing analysis of human skeletal muscle taken from m. vastus lateralis. AB - The relative content of myosin heavy chain (MHC) isoforms IIb, IIa and I in human skeletal muscle taken from the m. vastus lateralis of 30 healthy male subjects was analysed using mini-gel electrophoresis. Repeated electrophoretic gels utilizing the same methods were produced for all subjects and the determination of MHC protein bands was performed using a digital scanner and National Institutes of Health (NIH) Image software and laser densitometry. A comparison between the NIH Image processing technique and laser densitometry revealed differences of 6.47%, 6.35% and 6.84% between these measurement techniques for MHC-IIb, -IIa and -I isoforms, respectively. The percentage technical error of measurement (TEM%) between electrophoretic gels was shown to be 19.1%, 17.8% and 14.2%, with regard to percentage of occurrence of MHC-IIb, -IIa and -I isoforms respectively. The variation in electrophoretic gel analyses was shown to be 5.7%, 7.3% and 5.5%, with regard to the percentage of MHC-IIb, -IIa and -I isoforms respectively. Intra-class correlations comparing NIH Image and laser densitometry produced r values in the range 0.38-0.63. Comparisons between and within gel analyses produced r values in the range 0.59-0.94 and 0.93-0.98, respectively. Analyses of variance revealed no significant differences (P < 0.05) between analysis techniques, between gels or within gels for the measurement of MHC-IIb, IIa and -I isoforms. The inter-gel error between fibre subgroups was moderate for the two type-II MHC populations and less for type-I MHC; the intra-individual error in the measuring technique used for classifying the MHC-IIb, -IIa and -I protein bands was small. The results obtained in this investigation showed consistent trends which may reflect a false classification of the type-II MHC populations for the inter-gel and intra-individual analyses. The NIH Image software and digitizing process was shown to be a valid and reliable method for distinguishing between MHC protein bands of human skeletal tissue as separated by mini-gel electrophoretic techniques. PMID- 9202949 TI - Effects of different carbohydrate-electrolyte beverages on the appearance of ingested deuterium in body fluids during moderate exercise by humans in the heat. AB - To determine whether different forms of glucose (free and polymer) associated with sodium chloride influence the rate of water absorption during exercise in the heat, six men took part in five trials. Each trial included a passive heating session which resulted in a 2% loss of body mass, followed by 1h of treadmill exercise (at 50% of maximal oxygen uptake) in warm conditions (dry bulb temperature 35 degrees C, relative humidity 20%-30%). Immediately before exercise, the subjects were given either no fluid or a volume equal to 50% of the fluid previously lost (about 650 ml), chosen from among four D2O-labelled beverages: mineral water, a 6% glucose-electrolyte solution (GS), a 6% maltodextrin solution and a 6% maltodextrin-electrolyte solution. No significant differences were observed among these various beverages so far as temporal accumulation of deuterium in plasma, sweat and urine was concerned. During GS, the plasma volume was completely restored and the drifts of heart rate and rectal temperature were less marked than during other trials. These results would suggest that rehydration with GS was more efficient, probably because of an internal redistribution of water. The proportion of ingested water was twice as high in sweat as it was in urine. These findings may reflect the essential part played by circulatory adjustments in the transfer of plasma water into sweat and urine. PMID- 9202951 TI - Stroke volume response to cycle ergometry in trained and untrained older men. AB - The aims of this study were threefold: (1) to investigate the stroke volume (SV) response of trained older male cyclists [Cyclists: 65 (2.1) years, n = 10] during incremental cycle ergometry (20 W.min-1); (2) to determine the SV dynamics and total peripheral resistance response of untrained, but healthy and active older male controls [ CONTROLS: 66 (1.1) years; n = 10]; (3) to compare the maximum oxygen consumption (VO2max) and SV response of trained older male runners [Runners: 65 (3.4) years; n = 11] with that of age-matched Cyclists. Impedance cardiography was used to assess the response of cardiac output (CO), SV and total peripheral resistance to exercise involving cycle ergometry. The mean VO2max of the trained Cyclists [54 (1.6) ml.kg-1.min-1] was significantly higher (P < 0.05) than that of the Runner [48 (3.9) ml.kg-1.min-1], whereas both groups possessed a significantly higher VO2max than the CONTROLS [28 (1.3) ml.kg-1.min-1]. During exercise, at a heart rate of 90 beats.min-1, the SV of the Cyclists increased by 41%, that of the Runners increased by 47%, and that of the CONTROLS increased by 31%. However, the Cyclists' and Runners' SV response was significantly greater than that of the CONTROLS. The SV for cyclists and controls peaked at 30% of VO2max. This early increase in SV was a major factor underlying the increase in CO during exercise in both the trained and the untrained subjects. In addition, all three groups showed a significant decrease in total peripheral resistance throughout exercise. The finding that older male runners possessed a large exercise SV and high VO2max suggests that run training results in enhanced cardiovascular performance during cycle ergometry. PMID- 9202952 TI - Responses of plasma glutamine, free tryptophan and branched-chain amino acids to prolonged exercise after a regime designed to reduce muscle glycogen. AB - Prolonged exercise can elicit a reduction in the plasma glutamine concentration and an increase in the plasma concentration ratio of free tryptophan (FTrp) to branched-chain amino acids (BCAA). The purpose of this investigation was to study the effects of a 60-min bout of vigorous treadmill running with dietary manipulation on plasma concentrations of glutamine, FTrp and BCAA after an exercise and diet regime designed to reduce muscle glycogen. Seven male distance runners [mean (SD) age: 29.3 (2.1) years; VO2max: 62.7 (3.3) ml.kg-1.min-1] acted as subjects. Each undertook a regime designed to reduce muscle glycogen, then performed a 60-min treadmill run (75% VO2max) under two dietary conditions: after a 14-h fast (fasted) and after ingestion of a high carbohydrate meal (30 kJ.kg-1: 80% carbohydrate, 10% protein, 10% fat) 3 h before running (fed). Plasma concentrations of glutamine, FTrp, BCAA, free fatty acids (FFA), glycerol and glucose were measured 5 min before and 5 min after the run, under each dietary condition. Plasma glutamine did not change in response to exercise when fasted (P > 0.05), but increased when fed (P = 0.007). Plasma FTrp increased under both dietary conditions (P < 0.001), but the magnitude of this increase was greater when fasted than when fed (P < 0.001). Plasma BCAA did not change under either dietary condition (P < 0.05). Increases in the plasma FTrp/BCAA ratio reflected increases in plasma FFA and glycerol concentrations (P < 0.001; both dietary conditions) and these changes were all greater under fasted conditions when a fall in blood glucose concentration was observed (P = 0.007). These data emphasise the importance of dietary carbohydrate intake between repeated bouts of prolonged exercise on responses of plasma glutamine, FTrp and BCAA during subsequent exercise. PMID- 9202953 TI - The effect of gender on left ventricular function immediately after the wingate test. AB - The effect of gender on left ventricular systolic function and exercise haemodynamics in healthy young subjects was studied during 30-s all-out sudden strenuous dynamic exercise. A group of 22 men [19.3 (SD 1) years] 20 women [19.1 (SD 1) years] volunteered to participate in this study. Two-dimensional direct M mode and Doppler echocardiograph studies were performed with the subject in the sitting position. The Doppler examination of flow was located with continuous wave, interrogating ascending aorta measurements. The subjects completed the study without showing any electrocardiograph abnormalities. An interaction effect with stroke volume (P < 0.05) was characterized by a decrease in the men and an increase of stroke volume in the women. Cardiac output rose significantly (P < 0.05) up to 14.5 (SD 6) 1.min-1) for the men and 12.1 (SD 4) 1.min-1 for the women compared to the rest values [5.8 (SD 0.4) and 4.7 (SD 0.5) 1.min-1, respectively]. Flow velocity integral and acceleration time differed significantly between the two groups at rest (P < 0.05). During exercise these differences showed an interaction effect (P < 0.05). These results would indicate that normal men and women respond to sudden strenuous exercise by reducing their left ventricular systolic function, with a significantly greater decrease in women (P < 0.05). The gender differences in the haemodynamic responses during the present study, may, as suggested by others, be attributable to differences in energy metabolism. In addition, changes in Doppler parameters of aortic flow, haemodynamics and blood pressure responses during sudden strenuous exercise differed markedly from those seen before with endurance exercise. PMID- 9202954 TI - Validation of a dual energy X-ray absorptiometer: measurement of bone mass and soft tissue composition. AB - We investigated the reproducibility of total and regional body composition measurements performed on a dual energy X-ray absorptiometer (DXA). A group of 58 women aged 21-81 (mean 52.4) years was scanned twice with repositioning to determine intraobserver reproducibility of measurements of bone mineral density (BMD, g.cm-2), bone mineral content (BMC, g), lean mass (LM, kg) and fat mass (FM, kg) of the total body and of the major subregions of the body. In addition, the ability of the DXA machine to detect changes in LM and FM (simulated by placing 11.1 and 22.3 kg porcine lard on the body of 11 subjects) was examined. Coefficients of variations calculated from the root mean square averages of individual standard deviations were as follows (BMD, BMC, LM, FM) [corrected]: 1.4%, 1.1%, 1.4%, 1.7% (total body), 2.2%, 2.1%,-,- (head), 2.8%, 2.8%, 2.0%, 2.2% (trunk), 3.6%, 3.9%, 4.0%, 4.9% (arms), 2.7%, 1.3%, 2.6%, 2.8% (legs). Percentage fat (%fat) of exogenous lard was 81.3 (SD 3.5)% as assessed by the absorptiometer which corresponded well with the result of chemical analysis (82.8%). Estimated %fat of exogenous lard was not influenced by initial body mass or percentage body fat. Percentages of expected mean values with 11.1 kg lard placed on the body were 99.9 (SD 0.3) for body mass, 100.5 (SD 2.1) for LM, and 99.5 (SD 3.5) for FM. BMD was overestimated by 3.2% (P < 0.005) with 11.1 kg lard on the body. BMD as well as BMC increased significantly with 22.3 kg lard on the body (P < 0.005). The results showed that BMD, BMC, LM, and FM of the total body were precisely estimated by the DXA machine used. Regional measurements were less precise. Changes in total body soft tissue composition were precisely and accurately estimated. The lard placed on the body falsely affected BMD and BMC measurements. Changes in body mass could have a similar effect. PMID- 9202955 TI - Abnormal mucous cell phenotype induced by neutrophil elastase in hamster bronchi. AB - Bronchial mucous cell metaplasia (MCM) is a histologic component of chronic mucus hypersecretion. The hamster model of elastase-induced MCM appears to involve an irreversible conversion of Clara cells to mucous cells. The present study questioned whether the mucous cells seen in hamster bronchi exposed to neutrophil elastase produce and maintain a form of glycoconjugate secretory product different from that normally found in mucous cells or Clara cells. Ultrastructural cytochemistry using the gold-labeled lectin HPA revealed a difference in the cell surface and stored secretory granules of elastase-derived mucous cells compared to normal mucous cells and Clara cells at 3 weeks and 4 months following exposure. The results suggest that elastase irreversibly alters the glycoconjugate character of the Clara cells normally present so that they produce an abnormal form of mucus. Because secreted glycoconjugates can affect the rate of mucociliary clearance and receptor-mediated binding of microorganisms, this change in phenotype may be involved in the pathogenesis of diseases associated with chronic mucus hypersecretion in humans. PMID- 9202956 TI - Intratracheal aerosolization of endotoxin (LPS) in the rat: a comprehensive animal model to study adult (acute) respiratory distress syndrome. AB - The aim of the study was to extend existing evidence that intratracheal aerosolization of LPS may serve as a very relevant model to study ARDS. The authors investigated the sequence of pathogenic events reflected by changes in levels of tumor necrosis factor alpha (TNF alpha), surfactant-associated protein A (SP-A) in BAL fluid, in addition to cell count, edema formation, and respiratory function. Within 24 h following intratracheal aerosolization of LPS in the rat, ARDS could be diagnosed according to the lung injury score for patients. This score includes the extent of the inflammatory density on chest X rays, the severity of hypoxemia, the decline in lung compliance, and the level of PEEP (positive end expiratory pressure). In addition, other typical features of human ARDS appeared to be present in this model: (1) increased microvascular permeability reflected by edema, elevated levels of protein and of LDH, and increased numbers of PMNs in BAL fluid; (2) high levels of TNF alpha in BAL fluid preceding the appearance of PMNs; (3) changes in breathing pattern and a gradual development of respiratory failure with decreased compliance. SP-A levels in BAL fluid doubled within one hour after LPS administration, suggesting that this collectin may play a role in the immediate inflammatory response. Taken together, the findings presented here suggest that intratracheal LPS administration mimics the clinical development of ARDS very closely. PMID- 9202957 TI - Postexposure treatment with aminophylline protects against phosgene-induced acute lung injury. AB - Pretreatment with aminophylline has been shown to protect against various types of acute lung injury. Mechanisms responsible for protection are multifactorial but are thought to involve upregulation of cAMP. While previous studies focused on pretreatment, the present investigation examined post-treatment in rabbits following exposure to a lethal dose of the oxidant gas phosgene. Rabbits, 2-3 kg, were exposed to a cumulative dose of phosgene to attain a c x t exposure effect of 1500 ppm.min. Lungs were isolated in situ and perfused for 90-100 min after exposure with Krebs-Henseleit buffer at 40 mL/min. Pulmonary artery pressure (Ppa), tracheal pressure (Pt), and lung weight gain (lwg) were measured continuously. Leukotrienes C4/D4/E4 were measured in the perfusate every 20 min during perfusion. At the immediate conclusion of the experiment, lung tissue was frozen in liquid N2 and analyzed for reduced GSH, GSSG, cAMP, and lipid peroxidation (TBARS). Post-treatment with aminophylline 80-90 min after exposure significantly lowered Ppa, Pt, and lwg. Aminophylline significantly reduced TBARS and perfusate LTC4/D4/E4, and prevented phosgene-induced decreases in lung tissue cAMP. These data suggest that protective mechanisms observed with aminophylline involve decreased LTC4/D4/E4-mediated pulmonary capillary permeability and attenuated lipid peroxidation. Direct antipermeability effects of cAMP on cellular contraction may also be important in protection against phosgene-induced lung injury. PMID- 9202958 TI - Maturational changes in Na(+)-K+ pump activity in guinea pig airway smooth muscle. AB - The effect of maturation on the Na(+)-K+ pump of airway smooth muscle (ASM) was studied. Na(+)-K+ pump activity of tracheal smooth muscle from immature (1- to 2 week-old) and mature (10- to 12-week-old) guinea pigs was measured as the ouabain sensitive component of the resting membrane potential and as ouabain-sensitive 86Rb+ uptake (using 86Rb+ as a marker for K+). Maturation resulted in decreases in both the contribution of the Na(+)-K+ pump of the resting membrane potential and in ouabain-sensitive 86Rb+ uptake. 86Rb+ efflux from tracheal smooth muscle was similar in tissues from immature and mature guinea pigs, suggesting that maturation has no effect on K+ (or at least 86Rb+) permeability. Na+ and K+ contents of tracheal smooth muscle were estimated from the uptakes of 24Na+ and 86Rb+ at equilibrium. Maturation resulted in a decrease in Na+ content but had no effect on K+ content. Since intracellular Na+ is one of the principal determinants of Na(+)-K+ pump activity, these results suggest that maturation results in a decrease in Na(+)-K+ pump activity in ASM, which may be due to a decrease in Na+ content. PMID- 9202959 TI - Decrease in gamma-glutamyltransferase activity in rat type II cells exposed in vitro to hyperoxia: effects of the 21-aminosteroid U-74389G. AB - Although the effect of hyperoxia on antioxidant enzymes is well known, the effect of subtoxic levels of hyperoxia on gamma-glutamyltransferase (gamma-GT), involved in the degradation and uptake of extracellular GSH for intracellular GSH synthesis, is unknown. The aim of the study was to investigate (1) the effects of in vitro hyperoxia on gamma-GT activity of type II cells and (2) the effects of the lazaroid U-74389G and N-acetylcysteine (NAC) on the hyperoxia-induced changes in gamma-GT and antioxidant enzyme activities. At 48 h after isolation, rat type II cells were exposed for 2 days to air, 60% O2 or 85% O2 with or without 30 microM U-74389G or 100 microM NAC. After the exposure, the cells were harvested and assayed for superoxide dismutase (SOD), glutathione peroxidase (GPx), gamma GT activity, and GSH levels. In another series of experiments 85% O2-exposed cells, with or without U-74389G, were used for Northern blotting of gamma-GT mRNA. Exposure to 60% O2 decreased gamma-GT and GSH by -47 and -34%, respectively, while SOD and GPx activities remained unchanged. After 85% O2 exposure gamma-GT decreased by -55%, SOD and GPx increased by +55 and +87%, respectively, while GSH decreased by -35%. NAC treatment decreased gamma-GT activity by -42% in the air-exposed cells. After 60% O2, U-74389G led to significantly higher gamma-GT (+117%) and GSH (+26%) while NAC only led to higher GSH (+28%) compared to the oxygen-exposed cells not treated with NAC or U-74389G. After 85% O2 U-74389G increased gamma-GT, SOD, and GSH by +72, +58, and +68%, respectively, while NAC only increased SOD (+49%) and GSH (+26%) compared to the oxygen-exposed cells not treated with NAC or U-74389G. The 85% O2 exposure, with or without U-74389G, had no effect on gamma-GT mRNA levels. The results show that hyperoxia decreases rat type II cell gamma-GT activity in vitro. This effect was not related to an altered regulation at mRNA level and it was not associated with the hyperoxia-induced decrease in intracellular GSH, since restoration of the GSH levels by NAC did not restore gamma-GT activity. The lazaroid U-74389G with vitamin E-like properties effectively prevented the decrease in gamma-GT and GSH, so that direct inactivation of the membrane-bound gamma-GT by hyperoxia is the most likely mechanism. PMID- 9202961 TI - Normal heterophoric changes: 20 years' follow-up. AB - BACKGROUND: Evaluation of changes in the heterophoric condition of 100 normal individuals over a 20-year period. METHODS: A retrospective study was undertaken. Charts of 100 normal men were reviewed. Individual changes in the heterophoric status were recorded over a 20-year period. The average value of the phoria at age 18-22 years was compared with the average value at age 34-38 years. Measurements were taken for near and distance fixation. No one was heterotropic. Changes in convergence and accommodation were also calculated. RESULTS: A 0.9 +/- 1.7 prism diopter increase in esophoria for distance fixation and a 0.6 +/- 2.5 prism diopter increase in exophoria for near fixation were found. These changes were statistically significant (P < 0.001 and P < 0.02 respectively). The near point of convergence receded by 0.5 +/- 1.1 cm (P < 0.001), and a decrease in accommodation over time of 2.8 +/- 1.4 diopters was found (P < 0.001). CONCLUSION: An increase in esophoria for distance fixation and a exophoria for near fixation was found in a 20-year follow-up of 100 normal subjects. PMID- 9202960 TI - Central serous retinopathy associated with adrenocorticotrophic hormone therapy. A case report and a hypothesis. AB - BACKGROUND: Central serous retinopathy (CSR) has been linked by several authors to the therapeutic use of adrenocorticotrophic hormone (ACTH) or corticosteroids or to endogenous ACTH hypersecretion. Various pathogenic mechanisms have been suggested to explain this phenomenon; all relate to the steroids as the causative agents. CASE REPORT: A simultaneous, bilateral central serous retinopathy developed in a woman treated by intramuscular injections of a synthetic ACTH analog for arthritis. The condition resolved 2 months after stopping the use of this drug. DISCUSSION: Many investigators consider CSR to be either a disorder of the retinal pigment epithelium (RPE) ion-pump function or a result of choroidal vascular hyperpermeability. ACTH has a melanotrophic part, melanocyte-stimulating hormone (MSH), that may affect RPE cells, which are melanin-pigmented cells of neuroectoderm origin, and alter their ionic pumping properties. This is supported by evidence in the literature for the effect of MSH on animal RPE cells, as well as on other secreting epithelia. MSH is known to act through increasing intracellular cAMP levels. Based on the current concepts regarding the pathogenesis of CSR, two possible mechanisms for ACTH/MSH-associated CSR are suggested. In the first, MSH increases intracellular cAMP levels in RPE cells, thereby inducing pump dysfunction and a reversal of ionic current direction, leading to subretinal fluid accumulation. An alternative mechanism is based on the known ability of MSH to increase the permeability of blood-aqueous and blood brain barriers. It is hypothesized that MSH disrupts the outer blood-retinal barrier or causes leakage from choroidal vessels. PMID- 9202962 TI - Ab interno infrared laser trabecular ablation: preliminary short-term results in patients with open-angle glaucoma. AB - BACKGROUND: Outflow obstruction of trabecular drainage structures results in elevation of intraocular pressure (IOP) in various forms of chronic open-angle glaucoma. In vitro studies have shown that laser trabecular ablation (LTA) with opening of Schlemm's canal can be reproducibly performed by use of infrared lasers with minimal damage to collateral tissue structures. METHODS: In order to investigate the clinical applicability and efficacy of ab interno contact laser photoablation of trabecular meshwork in the surgical treatment of human glaucoma, we conducted a pilot study using an erbium: YAG laser (2.94 microns) with a quartz fiber endoprobe (320 microns core diameter) applying 10-20 single laser pulses (5-7 mJ) to the trabecular meshwork. RESULTS: Under goniocopic visualization trabecular tissue was photovaporized in eight patients with primary or secondary chronic open-angle glaucoma. Intraoperatively, moderate reflux bleeding occurred from laser-induced craters. No major intra- or postoperative complications occurred. Preoperative and postoperative gonioscopy of the treated area demonstrated successful removal of trabecular tissue. Mean IOP was reduced from 36.1 mmHg ot 21.3 mmHg at a limited follow-up of 3 months. CONCLUSION: With modifications this technique may have clinical potential in the surgical management of glaucoma. Prospective long-term studies with more patients are warranted to evaluate the outcome of LTA. PMID- 9202964 TI - Flecked chorioretinopathy associated with Birt-Hogg-Dube syndrome. AB - BACKGROUND: Birt-Hogg-Dube's syndrome is a rare skin disease characterized by multiple trichofibromas of the skin and polyps of the intestine. Ophthalmologic manifestations associated with the syndrome have not been reported in detail. CASE REPORTS AND METHODS: Two siblings suffering from Birt-Hogg-Dube syndrome were examined clinically. Electrooculography and electroretinography were performed according to international standards. Color fundus photographs were taken as well as fluorescein angiograms. The two patients showed multiple perifollicular fibromas and trichodiscomas of the skin of the head. Funduscopy and fluorescein angiography revealed a flecked chorioretinopathy in one patient with progressive constriction of visual fields and severely reduced electroretinographic responses. Ophthalmoscopy in his sister showed peripheral pigmentary changes with only minor functional abnormalities. CONCLUSION: These findings suggest that Birt-Hogg-Dube syndrome may be associated with a progressive flecked chorioretinopathy with constricted visual fields and that patients with the syndrome should undergo ophthalmological examination. PMID- 9202963 TI - Efficacy of argon laser trabeculoplasty: 3-year preliminary results of a prospective placebo-controlled study. AB - BACKGROUND: It was the purpose of this study to evaluate the efficacy of argon laser trabeculoplasty (ALT) in controlling intraocular pressure (IOP) and avoiding surgical intervention. We also investigated whether topically applied diclofenac sodium eye drops had an influence on the success rate of ALT compared with placebo eye drops. METHODS: The indication for ALT was progressive glaucoma uncontrolled by maximal tolerated medical therapy. Thirty-nine of 41 patients were available for follow-up after 1122 +/- 239 days. RESULTS: Twenty-one of 39 eyes failed during the first 3 years of follow-up for one or more reasons: 11 eyes required additional laser treatment and/or filtration surgery because of progressive visual field loss or unacceptably high IOP, 9 eyes failed to have a final IOP > or = 21 mmHg, and 7 eyes failed because of an increase in the number of the medications. This yields a success rate for ALT of 46% for 3-year follow up. There was no significant difference between diclofenac sodium- and placebo treated eyes concerning the success rate after 3 years (P > 0.05). CONCLUSION: We conclude that the use of ALT for the treatment of glaucoma is best reserved for cases in which medical avenues of treatment have been exhausted. PMID- 9202965 TI - Peripheral visual field loss after vitreous surgery for macular holes. AB - BACKGROUND: Vitreous surgery for Idiopathic macular holes can result in both anatomic closure of the hole and visual improvement. In some patients even normal visual acuity is achieved. Postoperative visual field loss is a newly recognized complication. This prospective study evaluates the frequency and significance of scotomas after vitrectomy with gas tamponade for stage I-IV macular holes. METHODS: Over a period of 10 months, a consecutive series of 30 patients and 31 eyes with macular holes underwent pre- and postoperative automatic perimetry (Octopus 07, 70 degrees) and macular perimetry (Octopus M1, 24 degrees) to characterize the pattern of visual field defects after vitrectomy with gas tamponade. Success rates were evaluated and complications were analyzed. RESULTS: Anatomic success after one surgical procedure was achieved in 85% of cases, visual improvement in 58%. Some 70.1% of patients had peripheral scotomas postoperatively; some of these were highly symptomatic and others were detected by visual field testing only. The most consistently affected areas were the temporal and lower periphery of the visual field. The central visual field, however, was not disturbed. CONCLUSION: Visual field loss after otherwise successful surgery for macular holes is an unexpectedly frequent and serious complication. The authors discuss various factors that may contribute to the postoperative scotomas. From the localization of the scotomas it seems most likely that they are caused by the persistent pressure of the gas bubble on the peripheral retina. Further investigations are necessary to confirm this hypothesis, and ways must be found to avoid this complication in order to be able to proceed with this otherwise promising new indication group for vitreous surgery. PMID- 9202966 TI - Hyperimmunoglobulinemia D in idiopathic retinal vasculitis. AB - BACKGROUND: We examined four patients who exhibited both idiopathic retinal vasculitis and elevated serum IgD levels. Uveitis caused by Behcet's disease is also associated with high levels of serum IgD. Therefore, the clinical features of these patients were investigated and the possible relationship between retinal vasculitis and elevated serum IgD was examined after undertaking a study of increased IgD levels in patients diagnosed with uveitis. METHODS: The study population was composed of 110 patients: 49 with Behcet's disease, 15 with sarcoidosis, 10 with Vogt-Koyanagi-Harada disease, and 36 with other forms of uveitis. IgD measurements were performed using modifications of the latex photometric immunoassay. Surface IgD (sIgD) expression in peripheral lymphocytes was determined by immunofluorescence, and the correlation between serum IgD levels and the percentage of sIgD-positive cells was examined. RESULTS: Twelve of the 110 patients had an elevated serum IgD. Eight of the 12 had Behcet's disease, and 4 were diagnosed with idiopathic retinal vasculitis. These 4 patients were HLA-A24+ females whose ages ranged from 8 to 25 years. A linear correlation between the serum IgD levels and the percentage of sIgD-positive cells was found. CONCLUSION: Hyperimmunoglobulinemia D state was found in Behcet's disease and idiopathic retinal vasculitis. These diseases may represent a new clinical entity characterized by signs of retinal vasculitis and hyperimmunoglobulinemia D that results from abnormal B cell activation and immune complex-mediated responses. PMID- 9202967 TI - Solutions containing miotic agents: effects on corneal transendothelial electrical potential difference. AB - BACKGROUND: Anterior chamber miotic solutions are widely used during anterior chamber surgery. We examined the effects of solutions containing miotic agents such as carbachol and/or acetylcholine on corneal endothelial pumping activity. METHODS: We monitored, in vitro, the transendothelial electrical potential difference of isolated rabbit corneal endothelial preparations. As controls, we used solutions without miotics. RESULTS: We found that a solution containing 55 mM acetylcholine and minimal amounts of salts (Miochol E) maintains transendothelial electrical potential difference some 30% above control levels for up to 4 h. Two other solutions, one including balanced salts and 0.55 mM carbachol (Miostat), the other a mixture of 0.19 mM carbachol and 55 mM acetylcholine plus minimal salts, are adequate to maintain the potential difference at control levels. Lastly, a solution with acetylcholine but without any salts (Miochol) greatly decreases the potential difference, to 30% of the control level, in 100 min. CONCLUSION: Our results indicate that: (1) 55 mM (1%) acetylcholine stimulates the endothelial electrical potential difference; (2) addition of 0.19 mM (0.003%) carbachol negates the stimulatory effect of acetylcholine; and (3) absence of electrolytes severely depresses the endothelial electrical activity. PMID- 9202968 TI - Effects of acetylcholine, carbachol, and mannitol on rabbit corneal endothelial function as assessed by corneal deturgescence. AB - BACKGROUND: Anterior chamber miotic solutions are widely used in ophthalmic surgery to induce pupillary contraction. We investigated whether the acetylcholine, carbachol, or mannitol present in perfusing solutions can affect corneal endothelial function. METHODS: Freshly dissected deepithelized rabbit corneas were mounted in a Dikstein-Maurice chamber at 36 degrees C. The endothelial sides were perfused with six solutions: (A) 55 mM (1%) acetylcholine Cl plus modified balanced salts; (B) control for A, with acetylcholine Cl replaced by sucrose; (C) 0.55 mM (0.01%) carbachol Cl plus balanced salts; (D) balanced salts solution (BS; control for C); (E) 3% mannitol plus modified balanced salts; and (F) modified balanced salts (control for E, with mannitol replaced by sucrose). Corneal thickness was followed for 3 h in each experiment. The effect of solution E did not differ from that of solution F. RESULTS: The carbachol-containing solution produced a small increase in corneal thickness compared to the control solution, while the acetylcholine-containing solution resulted in corneal thickness lower than that in control preparations. CONCLUSION: From these data, acetylcholine is harmless to the endothelium, and may actually stimulate its fluid pump mechanism. Carbachol, on the other hand, appears to have a detrimental effect. PMID- 9202969 TI - Effect of oxygen on relaxation of retinal pericytes by sodium nitroprusside. AB - BACKGROUND: This study addresses whether oxygen modulates the relaxation induced in retinal pericytes by sodium nitroprusside (SNP), a nitric oxide (NO) donor that stimulates the NO/guanylate cyclase pathway. METHODS: Bovine retinal pericytes were cultured on silicone. On the silicone surface, basal pericyte contractile tone induces wrinkles. Drug-induced changes in pericyte contractile tone were assessed by changes in the number of wrinkles. The effects of 100% nitrogen (hypoxia) and 100% oxygen (hyperoxia) were studied on: (a) the basal tone of quiescent pericytes, (b) the relaxation to 3 and 10 microM SNP or 1 microM forskolin, and (c) the recontraction that followed the washout of 3 microM SNP or 1 microM forskolin. RESULTS: Neither hypoxia nor hyperoxia had any apparent influence on pericyte basal tone, on forskolin-induced relaxation, or on pericyte recontraction after a forskolin-induced relaxation. In hypoxia, relaxations to SNP 3 microM (P < 0.05) and 10 microM (P < 0.01) were significantly more pronounced than in hyperoxia. Hypoxia also reduced the recontraction after an SNP-induced relaxation (P < 0.001). CONCLUSION: Oxygen modulates the relaxation of bovine retinal pericytes evoked by SNP (guanylate cyclase-mediated), but not the relaxation induced by forskolin (adenylate cyclase mediated). These results suggest that in the retinal capillary circulation an interaction between oxygen and the NO/guanylate cyclase pathway modulates pericyte tone, and thus potentially blood flow. PMID- 9202970 TI - Microvascular change of the anterior eye segment after tenotomy in the rabbit. AB - BACKGROUND: The microvascular changes secondary to anterior segment ischemia following tenotomy of the extraocular muscles have not been studied in the rabbit. METHODS: Using scanning electron microscopy of methyl-methacrylate ocular microvascular luminal castings, the anterior eye segment vasculature after tenotomy was documented and compared to that after occlusion of the bilateral long posterior ciliary arteries and that in the eyes that were not subjected to any surgical intervention. RESULTS: Five days and 1 week after the surgical intervention with tenotomy, microvascular change secondary to the anterior segment ischemia was not apparent, but 2 weeks after the tenotomy subtle evidence of ischemia such as new vessels in the iris was observed. Seven weeks after tenotomy, marked microvascular change was observed where corneal new vessels arose from the superior perilimbal arteries. In contrast, we found prominent microvascular changes 2 weeks after the occlusion of the long posterior ciliary arteries. CONCLUSIONS: Tenotomy of the rabbit eye causes microvascular change similar to that in occlusion of the long posterior ciliary arteries. This result suggests that the anterior ciliary artery of the rabbit contributes blood flow to the anterior eye segment and also has a stronger connection with the long posterior ciliary artery than previously reported. PMID- 9202972 TI - Regulation of ceruloplasmin by retinoic acid in the developing rat. AB - Ceruloplasmin (Cp), the major copper-binding protein in the plasma, is an acute phase protein with ferrioxidase activity. Both its oxidase activity and hepatic mRNA abundance increase during the developmental period. To test the possible role of retinoic acid, a derivative of vitamin A known to participate in cellular differentiation and development, on the developmental regulation of Cp, neonatal rat pups were injected with 2 micrograms 13-cis-retinoic acid (RA)/g body weight on postnatal day 1. Serum Cp activity and hepatic Cp mRNA were measured over the next 3 weeks in RA-treated and vehicle-treated controls. Serum Cp activity increased 2.5-fold 24 h after RA administration. However, hepatic Cp mRNA abundance was not elevated during this time period, suggesting that the action of RA on Cp activity was the result of post-transcriptional changes. PMID- 9202971 TI - Decorin and suramin inhibit ocular fibroblast collagen production. AB - BACKGROUND: The process of ocular wound healing with respect to glaucomatous filtering procedures is of current interest. Delaying this response in patients could possibly lead to more favorable surgical results. So far, only highly toxic antimetabolites have come into frequent clinical use. The possible efficacy of other groups of substances such as growth factor inhibitors has not yet been examined in vitro. METHODS: We exposed Tenon's capsule fibroblasts in tissue culture to various concentrations of decorin and suramin. The dose responses of type I and type III collagen to these inhibitors were measured using an ELISA type dot blot assay. Total cellular protein production was assayed by measuring the incorporation of tritiated leucine. RESULTS: At a concentration of 10 micrograms/ml, suramin reduced the collagen production by more than 80%. Decorin, at a concentration of 100 micrograms/ml, reduced type I collagen production by about 50% while type III collagen was reduced by 80%. At these concentrations, the total cellular protein production was not inhibited. CONCLUSIONS: Both suramin and decorin, which specifically inhibit the action of growth factors on target cells, reduce the production of collagen synthesis by Tenon's capsule fibroblasts. This is a specific effect, because total protein production is not influenced. This sets these substances apart from antimetabolites. Decorin and suramin may have clinical relevance in that they appear to interfere with ocular wound healing more specifically than the substances so far frequently used. PMID- 9202973 TI - Interaction between vitamin A and iron: effects of supplements in pregnancy. AB - Very few reports are available on the vitamin A status of Indian pregnant women in the rural population. The present study was carried out to assess the vitamin A status and the effect of vitamin A and iron supplementation in pregnancy on serum vitamin A levels in the rural women. It was observed that serum vitamin A levels showed a marginal decrease with advancing gestation. Though satisfactory as per WHO guidelines, supplementation with 60 mg of iron prevented this decrease. A higher dose of iron (120 mg) actually resulted in a similar vitamin A status as seen in vitamin A supplemented women. If this preliminary study can be confirmed it would appear that interactions between vitamin A and iron in pregnancy have a positive effect on the vitamin A nutriture. PMID- 9202975 TI - beta-Carotene and alpha-tocopherol inhibit the development of atherosclerotic lesions in hypercholesterolemic rabbits. AB - Male New Zealand White rabbits were made hypercholesterolemic by feeding an atherogenic diet (0.5% cholesterol, 3% peanut oil, and 3% coconut oil) with or without (control) antioxidants for 8 weeks. The antioxidant treatments were intravenous injection of beta-carotene (25 mg/kg/BW, twice weekly), dietary supplementation of alpha-tocopherol (0.5%), and a combination of both. Antioxidant treatments significantly increased plasma and LDL antioxidant levels in the above three groups. Intravenous injection of beta-carotene significantly decreased total and LDL cholesterol concentrations, thoracic atherosclerotic lesion area, and intimal thickness, but had no effects on LDL oxidation ex vivo as compared to control. Added dietary alpha-tocopherol significantly decreased the susceptibility of LDL to oxidation ex vivo, aortic atherosclerotic lesion area and intimal thickness, but had no effects on plasma cholesterol levels as compared to control. Combination of both antioxidants significantly decreased total and LDL cholesterol concentrations, susceptibility of LDL to oxidation ex vivo, as well as atherosclerotic lesion area and intimal thickness at aortic arch and thoracic aorta as compared to control, but not beta-carotene or alpha tocopherol groups. These data suggest that the antihypercholesterolemic effects of beta-carotene and antioxidant effects of alpha-tocopherol may benefit rabbits fed an atherogenic diet by inhibiting the development of atherosclerotic lesions. PMID- 9202974 TI - Lack of influence of a long-term high or low vitamin E diet on the oxidative DNA damage in the bone marrow of mice. AB - To investigate the influence of dietary vitamin E (VE) on DNA damage in bone marrow, we fed mice either a low VE diet (-VE), a basal VE diet (+30 mg VE/kg) or a high VE diet (+1000 mg VE/kg) for 50 weeks. DNA damage was evaluated by two cytogenetic methods: micronucleus (MN) assay using peripheral blood, and examination of sister chromatid exchanges (SCE) in bone marrow cells. The MN assay was performed periodically from 6 to 50 weeks, and showed that the incidence of reticulocytes containing MNs did not increase in the low VE diet group, and did not decrease in the high VE diet group. SCE assay done at 50 weeks also showed no difference among the VE diet groups. VE in bone marrow was markedly lower in the low VE diet group and higher in the high VE diet group compared to that in the basal VE diet group at 6 weeks. The VE at 50 weeks was not markedly different from that at 6 weeks. Corresponding to the changes in the VE, lipid peroxide in bone marrow was higher in the low VE diet group, but was not lower in the high VE diet group. Glutathione and vitamin C in the bone marrow, which were about 100-1000 times higher than that of VE, were not different among the groups. PMID- 9202976 TI - Effects of methylcobalamin on the proliferation of androgen-sensitive or estrogen sensitive malignant cells in culture and in vivo. AB - Methylcobalamin is one of the coenzymatically active cobalamin derivates and required for the activity of the cytoplasmic enzyme methionine synthetase catalyzing the methylation of homocysteine into methionine. The effect of methylcobalamin on the proliferation of malignant cells has been examined. Methylcobalamin inhibited the proliferation of androgen-sensitive SC-3 cells (a cloned cell line from Shionogi mouse mammary tumor, SC115) in culture at the concentration of 100-300 micrograms/ml. An inhibitory activity of methylcobalamin on the proliferation was also observed in other cell lines (estrogen-sensitive B 1F cells from mouse Leydig cell tumor and MCF-7 cells from human mammary tumor) at the concentration of 500 micrograms/ml. Moreover, large doses of methylcobalamin injected intraperitoneally (100 mg/kg body weight/day) were non toxic and suppressed the tumor growth of SC115 and B-1F cells in mice fed a vitamin B12 deficient diet. These results show that methylcobalamin inhibits the proliferation of malignant cells in culture and in vivo and propose the possibility of methylcobalamin as a candidate of potentially useful agents for the treatment for some malignant tumors. PMID- 9202977 TI - Folate and vitamin B12 status in a group of preschool children. AB - An adequate intake of folates and vitamin B12 is essential for the rapid growth rates characteristic of infancy. However, little information exists on the prevalence of deficiencies of these nutrients in preschool children. The status of these vitamins was therefore evaluated in a group of 79 children between 2 and 6 years of age. A 5 day dietary record (including a Sunday) was kept for all subjects. All food taken at day care centres was monitored using "Precise individual weighing" and recorded by trained personnel. Measurements were made of serum and erythrocyte folate levels, vitamin B12 levels, number of red blood cells, haemoglobin and haematocrit levels, and mean corpuscular volume. Though the mean intake of folic acid surpassed recommended levels for this age group 31.4% of the subjects showed intakes below those recommended. 7.7% of the subjects showed serum folate levels between 3 and 6 ng/mL, values which indicate a moderate deficiency of this vitamin. A correlation was found between folate intake and serum folate levels r = 0.3654 (P < 0.01). Vitamin B12 intake was 438% that recommended. Only 2.9% of the subjects showed vitamin B12 intake below recommended and none showed serum values below 150 pg/ml, the lower normal limit below which deficiency is considered to exist. CONCLUSION: Amongst preschool children, folate deficiency is probably much more common than vitamin B12 deficiency. However, its incidence is low, and much lower than that observed in other age groups. PMID- 9202978 TI - Influence of the number of meals taken per day on cardiovascular risk factors and the energy and nutrient intakes of a group of elderly people. AB - The aim of this investigation was to analyse the influence of the number of meals per day on a range of cardiovascular risk factors and on the energy and nutrient intakes of a group of elderly people. The participants in this study were 150 elderly people (64 men and 86 women) from Madrid. Food intake was followed over a period of 5 days. "Precise individual weighing" was used to determine the intake of institutionalized subjects (n = 58) whilst "food intake records" were used to register the same for independent subjects (n = 92). The nutrient and energy intake of the studied population was then determined from these data. The number of meals taken was also recorded. Serum cholesterol and triacylglycerol levels were determined using enzymatic methods. In this population, the meal most frequently omitted was breakfast. No subject took only one meal per day, though 7.4% took only two. 56.6% took three meals and 36% took four. No subject took more than four meals per day. As the number of meals taken increased, so too the covering of theoretical energy expenditure, and the intakes of a range of nutrients became closer to those recommended e.g. proteins, fibre, vitamin C, thiamin, riboflavin, calcium, magnesium and iodine. As the number of meals taken per day increased, carbohydrate intake (in g/1000 Kcal and as % of energy) also increased, and approached recommended levels more closely. As observed in other studies, blood cholesterol levels were seen to be negatively correlated with increasing number of meals (r = -0.2297, p < 0.05). Further, those subjects who distributed their food intake more evenly throughout the day showed lower serum cholesterol (p < 0.05). VLDL-cholesterol (p < 0.05) and triacylglycerol levels (p < 0.05). The results favour the distribution of energy intake over the day as a method of improving nutritional status and as a factor that might improve blood lipid profiles. PMID- 9202979 TI - Health and dietary characteristics of supplement users in an elderly population. AB - The purpose of this study was to investigate the association of health and dietary characteristics with the use of vitamin and mineral supplements in community-dwelling, cognitively intact elders aged in their 60s (n = 89), 80s (n = 92), and 100s (n = 76) who resided in Georgia in the southeastern United States. Elders who were physically active (p = 0.008), had stomach problems (p = 0.042), or used arthritis medication (p = 0.015) were more likely to take a nutritional supplement than elders without these characteristics. Physically active elders were more likely to take calcium (p = 0.004), vitamin E (p = 0.022), and vitamin C (p = 0.046) than non-physically active elders. Compared to non-users, supplement users were also more likely to comply with nutritional health seeking behaviors such as avoiding too much salt, fat, cholesterol, sugar, caffeine, and eating enough fiber, vitamins and minerals from food or supplements, and calcium in foods or supplements. The observation that the use of certain vitamin or mineral supplements is associated with dietary fat intakes, dietary protein intakes, and patterns of alcohol, decaffeinated coffee, and tea consumption suggests that supplement use is one of a cluster of health behaviors. Thus, it may be important that future investigations concerning the impact of supplement use on diseases, such as heart disease or cancer, control for the effects of dietary patterns and physical activity. PMID- 9202981 TI - Effect of dietary fish oil on plasma lipoprotein cholesterol in rats fed a diet enriched in cholesterol and sucrose. AB - The aim of this study was to examine the effect of dietary fish oil on plasma lipoprotein cholesterol levels in rats fed different carbohydrate sources. Male Wistar rats fed a soy bean oil diet or a fish oil diet containing 0.5% cholesterol were studied for 7 weeks. Corn starch or sucrose were used as carbohydrate sources in the experimental diet. Fish oil supplementation significantly (p < 0.05) decreased plasma VLDL (very low density lipoprotein) cholesterol in rats fed a diet containing corn starch. However, there was no significant difference in plasma total cholesterol and LDL (low density lipoprotein) cholesterol in rats fed a corn starch diet with fish oil treatment. In the experiment with sucrose, significantly (p < 0.05) decreased plasma total cholesterol, LDL cholesterol and VLDL cholesterol were observed in rats fed a fish oil diet. Although higher plasma total and VLDL cholesterol levels were found in rats fed the sucrose diet when compared with those fed the corn starch diet, no significant difference between the corn starch group and the sucrose group was observed in rats after fish oil treatment. Results from the present study suggest that the carbohydrate source might play an important role in the regulation of plasma lipoprotein metabolism in rats fed fish oil. PMID- 9202980 TI - Effects of protein source and amino acid supplementation on plasma cholesterol in guinea pigs. AB - The effects of three dietary protein treatments were compared on cholesterol content of plasma lipoprotein fractions and oxidative status of liver lipids in adult guinea pigs. All diets were adequate in soluble dietary fiber and well balanced in fatty acids providing 30% of total energy. After seven weeks dietary treatments, casein compared to soy protein increased cholesterol in a sub fraction of LDL (low density lipoprotein) with larger molecular weight and in a combination of this sub-fraction of LDL plus VLDL (very low density lipoprotein) taken together. Supplementation of casein diet with glycine, alanine, arginine and cystine tended to decrease cholesterol in the sub-fraction of LDL with larger molecular weight. There was no effect of dietary treatments on thiobarbituric acid reactive substances in lipids extracted from guinea pig liver likely due to the very high vitamin E and C content of the diets. In addition to counteracting the serum cholesterol elevating effects of dietary cholesterol soy protein also appears to attenuate the hypercholesterolemic effects of dietary saturated fatty acids. PMID- 9202983 TI - Computer-aided semen analysis (CASA) 10 years later: a test-bed for the European scientific andrological community. PMID- 9202982 TI - The effect of calcium gluconate and other calcium supplements as a dietary calcium source on magnesium absorption in rats. AB - The effects of commercially available calcium supplements (calcium carbonate, calcium gluconate, oyster shell preparation and bovine bone preparation) and gluconic acid on the absorption of calcium and magnesium were evaluated for 30 days in male Wistar rats. There were no differences in the apparent absorption ratio of calcium among rats fed each calcium supplement; however, the rats fed the calcium gluconate diet had a higher apparent absorption ratio of magnesium than the rats fed the other calcium supplements. Dietary gluconic acid also more markedly stimulated magnesium absorption than the calcium carbonate diet, and the bone (femur and tibia) magnesium contents of rats fed the gluconic acid diet were significantly higher than those of the rats fed the calcium carbonate diet. Furthermore, the weight of cecal tissue and the concentrations of acetic acid and butyric acid in cecal digesta of rats fed the calcium gluconate diet or the gluconic acid diet were significantly increased. We speculate that the stimulation of magnesium absorption in rats fed the calcium gluconate diet is a result of the gluconic acid component and the effect of gluconic acid on magnesium absorption probably results from cecal hypertrophy, magnesium solubility in the large intestine and the effects of volatile fatty acids on magnesium absorption. PMID- 9202984 TI - Localization of integrin beta 1, alpha 1, alpha 5 and alpha 9 subunits in the rat testis. AB - Very late activation (VLA, beta 1; alpha 1; alpha 5, alpha 9) integrins were studied by immunoblotting and immunohistochemistry in the testes of sexually mature rats. All integrin subunits were present in membrane fractions of homogenized testes. Immunohistochemistry revealed that the anti beta 1 antibody recognized peritubular cells and the basement membrane of blood vessels. Immunoreactivity was also demonstrated in the lamina propria, basement membrane, and the basal cytoplasm of Sertoli cells. In elongating spermatids, beta 1 integrin was localized to the acrosome. The alpha 1 subunit was expressed in peritubular cells and in the lamina propria. In the adluminal compartment, round spermatids were stained diffusely for the alpha 1 subunit. Immunoreactivity for alpha 1 integrin was found additionally in the acrosomes of elongating spermatids shortly before their release into the seminiferous tubule lumen. The alpha 5 subunit was expressed in the acrosomes of elongating spermatids as well as in their distal cytoplasm during stages III-VI; the cytoplasmic lobes of elongate spermatids and/or residual bodies also appeared to be immunostained in seminiferous tubules at stages VII-VIII. The alpha 9 subunit was immunolocalized only in the basement membrane and in peritubular cells. These data suggest that integrins are involved in spermatogenesis, in particular in the process of spermatid maturation. PMID- 9202985 TI - Influence of fibronectin on the motility of human spermatozoa. AB - The objective of the present study was to scrutinize the concentration of seminal fibronectin and the potential effects of exogenous fibronectin on human sperm motility. In addition, variability in the localization of fibronectin on human spermatozoa from andrological patients was studied, at both the light and electron microscopic levels. A total of 58 freshly ejaculated semen samples from patients attending for infertility treatment were submitted to sperm motility analysis and ELISA quantification of seminal plasma and cell-bound fibronectin. Immunofluorescence and immunoelectron microscopy revealed a relatively broad distribution pattern of fibronectin immunoreactivity on sperm heads and testicular spermatids. Addition of a fibronectin antiserum to vital spermatozoa in vitro at a moderate dilution (1:50) resulted in a significant increase in sperm motility. Purified plasma fibronectin, added at various concentrations to a preparation of live spermatozoa, was found to inhibit sperm motility in a dose dependent manner. At concentrations from 0.18 to 0.5 mg fibronectin per ml ejaculate, no motile spermatozoa were recorded. Seminal plasma fibronectin ranged between 0.8 and 1000 micrograms/ml in infertility patients. There was a significant inverse correlation between sperm motility and seminal fibronectin in patients with oligo-astheno-teratozoospermia. In a preliminary study in patients with varicocele or hypogonadism, no such correlation was found. PMID- 9202986 TI - Enlarged spectrum of seminological diagnoses using the resazurin colour reaction; a spectrophotometric application. AB - Two sets of investigations were carried out in the present study to establish whether, in semen samples processed for the resazurin reaction (RT), the colour change and its intensity were informative for both the quality and quantity of spermatozoa. First, the results of RT were read visually using single semen samples from 42 consecutive men from infertile couples. In the second step, the results were additionally read using a spectrophotometer. Single semen samples obtained from another 87 consecutive men from infertile couples and 12 men from fertile couples were analysed in this step. Visual analysis, which involved grading against and RT colour chart, revealed a significant positive correlation between the RT results and the quantity and quality of spermatozoa. Azoospermia was diagnosed by grade number 1 (dark purple colour), while the highest grades 8 11 (colours dark red to pink) represented a homogeneous group of patients with the highest quality of spermatozoa (> 60% motility) and quantity (> 60 x 10(6)/ml ejaculate). Normal parameters of spermatozoa, according to WHO, appeared with a high predominance (89-100% of cases) in this group and were present in most of the samples. Thus, the grades 8-11 might represent semen samples with optimal fertility potential. Spectrophotometric analysis of RT revealed that visual characteristics corresponded closely to the values for optical density (OD) of the samples. The OD for normozoospermic samples ranged between 1.48 and 1.84; for oligozoospermia between 1.2 and 1.48; for azoospermia between 0.98 and 1.1. The OD for samples with putative optimal fertility potential ranged between 1.6 and 1.84. RT evaluation using a spectrophotometer may therefore provide a tool for obtaining a wider spectrum of seminological diagnoses than classic microscopic examination, especially for distinguishing men with probable optimal fertility potential. PMID- 9202987 TI - Antisperm antibodies in prepubertal boys treated with chemotherapy for malignant or non-malignant diseases and in boys with genital tract abnormalities. AB - In several childhood diseases which have the ensuing risk of infertility in adult life because of direct hypothalamic-pituitary-testicular axis involvement, or as a consequence of therapeutic toxicity, the role of antisperm antibodies (ASA) is rarely addressed. The aim of this study was to investigate the occurrence of ASA in a large prepubertal male population (aged 1.2-13 years) consisting of three groups: Group I, 52 patients affected by malignant diseases (lymphoblastic leukaemia, malignant lymphoma, or Wilm's tumour, n = 42), or by nephrotic syndrome (n = 10); Group II, 212 patients with either genital tract abnormalities (cryptorchidism, inguinal hernia, funicular torsion or hypospadias, n = 202), or cystic fibrosis (n = 10); Group III: 100 age-matched normal boys. Group I and II patients were investigated at diagnosis and during or after treatment (drug, radiation or surgical therapy). Group III was used as controls. ASA were detected in sera by the Tray Agglutination Test (TAT) and indirect IgG, IgA and IgM immunobead tests (iIBT). All normal boys were ASA-negative using both tests. Twenty-six out of the 264 patients (9.8%) in Groups I and II were ASA-positive: 23 (8.7%) patients had a positive TAT with a titre of 1:32 to 1:128, whilst 14 (5.3%) had IgG-ASA after iIBT. Eleven patients (4.1%) were ASA-positive in both tests. Of the 26 ASA-positive boys, 24 had genital tract abnormalities (cryptorchidism, testicular torsion, hypospadias) and two had leukaemia with testicular infiltration. Treatment did not modify antibody positivity. Our data confirm that ASA can occur in prepubertal boys, mostly among cases with urogenital pathology, but that it is rare among other cases. Therefore autoimmune reaction against spermatozoa is another factor that should be considered in the evaluation of several conditions in childhood involving reproductive tract alteration and potential impairment of the blood testis (Sertoli cell) barrier. PMID- 9202988 TI - Does the gonadotrophic axis play a role in the pathogenesis of Sertoli-cell-only syndrome? AB - Infertile men with Sertoli-cell-only syndrome (SCO) have highly elevated serum FSH immunoactivity related to the degree of histological damage. The activity that serum FSH exerts at the target site depends on its glycosylation pattern and FSH receptor (FSHR) function. Either could be impaired, leading to failure of spermatogenesis. The aim of the present investigation was to study bioactivity and the glycosylation pattern of serum FSH and the occurrence of mutations in the FSH receptor in infertile patients with SCO compared to normal men. Blood was taken from 19 patients with bilateral testicular focal or complete SCO and eight normozoospermic controls. FSH bioactivity in serum was measured using an in-vitro FSH bioassay based on recombinant rat FSHR. The glycosylation pattern of serum FSH was determined by concanavalin A chromatography. Inhibin B was determined in serum using a recently available assay. Genomic DNA extracted from blood lymphocytes was amplified by PCR using primers specific for the FSHR and screened by single-stranded conformation polymorphism gel electrophoresis. Men with SCO showed significantly higher FSH in-vitro bioactivity (34.9 +/- 5.0 IU/l) than controls (9.6 +/- 0.8 IU/l: p < 0.01), as well as significantly elevated FSH immunoactivity (14.9 +/- 1.7 IU/l) compared to controls (3.1 +/- 0.5; p < 0.01). Immunoactivity of serum FSH was correlated with in-vitro bioactivity (r = 0.9; p < 0.001) and was related to the degree of testicular damage (proportion of SCO tubules) (ANOVA: p < 0.001) and total testicular volume (r = -0.76; p < 0.01). An inverse relationship between serum FSH and inhibin B levels (r = -0.93; p < 0.001) was found. In the serum of SCO patients a slight increase in less glycosylated FSH isoforms was found (6.7 +/- 0.6% versus 3.6 +/- 0.3%; p < 0.05). No mutations of the FSHR were observed in SCO patients. We conclude that the spermatogenic failure observed in infertile patients with SCO histology and elevated FSH serum levels can be explained neither by a change in FSH bioactivity nor by mutations in the FSHR. The slight change in the FSH glycosylation pattern is probably related to higher hormonal secretion rates in SCO patients. The inverse relationship between serum FSH and inhibin B points to an intact endocrine testicular-pituitary circuit responsible for the compensatory increase of FSH in SCO. PMID- 9202990 TI - Improved preservation of ultrastructural morphology in human spermatozoa using betaine in the primary fixative. AB - Improved preservation of the ultrastructural morphology of human spermatozoa was obtained by filtration onto a Millipore filter (0.45 micron pore size) followed by fixation for 1 h in a fixative containing 3% glutaraldehyde, 1% formaldehyde and 21 mM betaine in Hepes or cacodylate buffer. Betaine raised fixative osmolality only slightly but markedly improved the ultrastructural appearance of the spermatozoa; it is known to function as an osmoprotectant in other situations, and may protect the cells from osmotic damage during the initial stages of fixation. The method is suitable for oligozoospermic samples as they are concentrated when the seminal plasma passes through the filter. Utilizing fixation after swim-up, it is suitable also for highly viscous semen. The fixative gave equally good preservation of sperm heads and tails and can be stored prior to use. The Millipore filter did not dissolve until placed in acetone prior to embedding, simplifying the exchange of solutions during processing. Improved preservation of rat epididymal ultrastructure was obtained by addition of betaine to the fixative of Ito & Karnovsky (1968), using immersion fixation. PMID- 9202989 TI - Intracytoplasmic sperm injection using surgically retrieved epididymal and testicular spermatozoa in cases of obstructive and non-obstructive azoospermia. AB - This was a retrospective study of 115 patients who underwent 124 cycles of ICSI using surgically retrieved spermatozoa. The objective was to compare the results of ICSI in patients with obstructive azoospermia using epididymal spermatozoa (36 cycles) or testicular spermatozoa (58 cycles) with ICSI in patients with non obstructive azoospermia using testicular spermatozoa (30 cycles). When epididymal spermatozoa were used for ICSI, the fertilization rate per injected metaphase-II oocyte and the clinical pregnancy rate per ICSI cycle were 60.4 and 25%, respectively. When testicular spermatozoa were used in obstructive cases, the fertilization rate and pregnancy rate were 57.9 and 34.5%. In non-obstructive cases the fertilization and pregnancy rates were 41.2 and 16.6%. When patients with obstructive azoospermia were regrouped according to the cause of obstruction, the fertilization and pregnancy rates were 59.1 and 35.1% in acquired obstruction and 58.7 and 24.3% in congenital obstruction. The fertilization and pregnancy rates were not statistically different (p > 0.05) when testicular or epididymal spermatozoa were used in obstructive cases; neither was statistically different (p > 0.05) when compared in patients with congenital and acquired obstruction. On the other hand, the fertilization and pregnancy rates in cases with non-obstructive azoospermia were significantly lower (p < 0.05) than in obstructive cases. PMID- 9202991 TI - Atrial natriuretic peptide, brain natriuretic peptide and c-type natriuretic peptide: effects on testicular microcirculation and immunohistochemical localization. AB - The effect of local injection of atrial (ANP), brain (BNP) and C-type (CNP) natriuretic peptides and an ANP antagonist (HS-142-1) on testicular microcirculation and vasomotion was studied using laser Doppler flowmetry. The natriuretic peptides were also localized immunohistochemically within the testis. ANP, BNP-32, CNP-22 and CNP-53 all caused a dose-related increase in testicular blood flow. The effect of ANP was blocked by concomitant injection of the ANP antagonist. Immunoreactive (ir) CNP and ir BNP were found in Leydig cells whereas in ANP was observed in the seminiferous tubules. It is suggested that the natriuretic peptides could play a role in local regulation of the testicular microcirculation. PMID- 9202992 TI - Alpha oscillations in brain functioning: an integrative theory. AB - The old concept stating that EEG alpha (10-Hz) activity reflects passive or idling states of the brain is giving way to modern views of 10-Hz oscillations in relation to diverse brain functions comprising sensory, motor, and memory processes: (1) Spontaneous alpha activity is not pure noise as shown by methods of chaos analysis. (2) Evoked alpha oscillations patterns (precisely time-locked to a stimulus; duration approx. 200-300 ms) depend on the modality of stimulation and the recording site. (3) Induced alpha oscillations are initiated by, but not closely time-locked to a stimulus. (4) 10-Hz oscillations are recorded in nervous systems of different complexities, from the human brain to isolated ganglia of invertebrates. The neural origins of 10-Hz oscillations are demonstrated by recordings at the cellular level. (5) Rather than trying to locate a unique alpha generator, it is preferable to assume that a 'diffuse and distributed alpha system' exists. A particular support for this hypothesis is given by stimulus dependent hippocampal alpha responses in the cat brain. (6) The major physiological meaning of 10-Hz oscillations may be comparable to the putative universal role of gamma responses in brain signaling. PMID- 9202993 TI - Alpha rhythms as physiological and abnormal phenomena. AB - There are three physiological alpha rhythms in mature healthy humans: (a) the classical posterior alpha; (b) the Rolandic mu rhythm and (c) the midtemporal 'third rhythm'. The classical posterior alpha rhythm develops out of a 4/s rhythm appearing at age 4 months and gradually reaches the alpha frequency band around age 3 years. The mature frequency around 10/s is subject to subtle physiological changes and grossly decelerates in the face of pathology. No posterior alpha rhythm may be detectable in a minority of healthy adults with an inherited low voltage fast EEG. One is tempted to speculate that these individuals may have a hidden alpha rhythm in neuronal level and defective mechanisms of synchronization. Alpha blocking with visual stimuli (eye opening) is a classical response; responses to mental stimuli (mental arithmetic) are inconsistent, presumably due to the involvement of higher cognitive functions. The Rolandic my rhythm is found with scalp EEG in a minority of subjects but there is good reason to presume that all healthy adults have this rhythm. A particularly powerful mu rhythm reaches the scalp but this could be also an indicator of a mild CNS dysfunction. There is even a relationship between mu rhythm and the central spike activity in children with benign Rolandic epilepsy. The midtemporal third rhythm is not detectable in the scalp EEG unless there are local bone defects. Its functional significance is debatable; its blocking responses encompass various higher cognitive tasks and are inconsistent; responses to auditory stimuli do occur but appear to be of secondary significance. This rhythm arises from midtemporal structures which by far exceed the borders of the auditory cortex. Abnormal rhythmical alpha activity-above all the alpha coma in life-threatening cerebral anoxia -is discussed in order to deepen our understanding of the physiological alpha rhythms. Severe cortical de-afferentation may give rise to cortical autorhythmicity-either in alpha frequency or in other frequency bands. Physiological alpha rhythms are likely to have closer relationships to 'events' than one might have thought earlier. The demonstration of event-related desynchronization and synchronization (in Pfurtscheller's work) clearly underscores this view. PMID- 9202994 TI - Magnetoencephalographic cortical rhythms. AB - We have characterized the magnetic 10- and 20-Hz rhythms recorded with a whole scalp neuromagnetometer during different conditions. Sources of the posterior 10 Hz (alpha) rhythm clustered mainly around the parieto-occipital sulcus and, to a lesser extent, around the calcarine sulci, with several generators. Temporal Spectral Evolution (TSE) analysis, used to follow event-related changes in the different frequency bands, showed strong dampening of the alpha within 200 ms after the appearance of a visual stimulus and also during visual imagery. Suppression was often followed by a rebound above the baseline level. The rolandic mu rhythm consisted of 10- and 20-Hz components with different reactivity and source locations. The 10-Hz component seems to be mainly somatosensory in origin whereas the 20-Hz signal also receives contributions from the motor cortex, and even shows 'motorotopy' in its reactivity: the source locations depend in a somatotopical manner on the site of the moving body part. The frequency composition of the posterior spontaneous activity was disturbed in patients with small infarcts of the medial thalamus. It is shown with simulations that a surprisingly small number of synchronized cortical neurons could generate the major part of the recorded oscillatory signal. Finally, some clarifications are suggested to the terminology of brain rhythms. PMID- 9202995 TI - Study of human occipital alpha rhythm: the alphon hypothesis and alpha suppression. AB - Alpha rhythm of the parieto-occipital area is comprised of a parade of short lived cortical excitations (alphons), each of which exhibits oscillations having a stable period within the alpha bandwidth. Strong alpha rhythm is produced by alphons extending over a larger cortical area, although an enhanced cortical current density may also contribute. Local suppression of alpha rhythm indicates when specific cortical areas become engaged in sensory or cognitive functions. Examples are provided for mental imagery, visual memory, auditory memory, and silent rhythming. PMID- 9202997 TI - Spatio-temporal dynamics of alpha brain electric fields, and cognitive modes. AB - Brain electric activity is viewed as sequences of momentary maps of potential distribution. Frequency-domain source modeling, estimation of the complexity of the trajectory of the mapped brain field distribution in state space, and microstate parsing were used as analysis tools. Input-presentation as well as task-free (spontaneous thought) data collection paradigms were employed. We found: Alpha EEG field strength is more affected by visualizing mentation than by abstract mentation, both input-driven as well as self-generated. There are different neuronal populations and brain locations of the electric generators for different temporal frequencies of the brain field. Different alpha frequencies execute different brain functions as revealed by canonical correlations with mentation profiles. Different modes of mentation engage the same temporal frequencies at different brain locations. The basic structure of alpha electric fields implies inhomogeneity over time-alpha consists of concatenated global microstates in the sub-second range, characterized by quasi-stable field topographics, and rapid transitions between the microstates. In general, brain activity is strongly discontinuous, indicating that parsing into field landscape defined microstates is appropriate. Different modes of spontaneous and induced mentation are associated with different brain electric microstates; these are proposed as candidates for psychophysiological 'atoms of thought'. PMID- 9202996 TI - The possible meaning of the upper and lower alpha frequency ranges for cognitive and creative tasks. AB - This study is aimed at verifying the functional independence of two frequency bands within the alpha range. It is based on experiments that examined the role of these two hands with regard to the amount of local electrogenesis (amplitude) and the cooperation of brain areas (coherence) in mental tasks concerning: (1) visual perception and imagery; (2) listening to and composing music; (3) verbal and visual creativity; and (4) aspects of mood. In all experiments EEG were recorded for at least 1 min during each task, separated one from another by at rest periods of at least equal lengths. EEG electrodes were pasted according to the 10/20 system (averaged ear lobes as reference). After FFT power was calculated for all 19 electrodes, coherence was estimated for all possible electrode pairs (i.e. 171). This was done for six frequency ranges between 1.5 and 31.5 Hz, the alpha range having been divided into two (7.5-9 Hz and 9.5-12.5 Hz). The spectral parameters obtained during each task were compared with those of the merged EEG at rest, significant changes (P < or = 0.01-P < or = 0.05) were entered into schematic maps of the brain. Generally, fewer differences were found for amplitude than for coherence. In all four tasks concerning visual perception the clearest differences were found in single person studies. But also in group studies more or less distinct differences were found between alpha 1 and 2. Also in the series with music the two alpha bands did not behave uniformly, nor were uniform features found in the two series of musically trained and untrained subjects. Distinct discrepancies were also found in a verbal and visual imagery task. With respect to mood, only elevated mood was correlated with a decrease of coherence in alpha 2 and an increase of amplitude in alpha 1. This study though hinting at a different functional significance of these two alpha bands, however, does not allow to draw any conclusions as to their distinct functional meanings. Generally, the long-term coherence changes observed under these different mental tasks support the idea that part of information processing in the brain is reflected by the EEG. Structural peculiarities and microelectrode recordings of the cortex support this conclusion. PMID- 9202998 TI - How reproducible is the topographical distribution of EEG amplitude? AB - If topographical EEG is to be a useful tool for localising cerebral processes, then the results of the same, or closely similar experiments, using different samples should yield similar results. Although the reliability of EEG is well established in other ways, there is little available data on the reproducibility of EEG topography across experiments. The aim of this study was to determine the reproducibility of topographical EEG by comparing the results of two independently conducted experiments. EEG was recorded during an Eyes Open baseline and a motor task condition (the Luria finger opposition task) in two independent samples of healthy subjects. EEG was recorded in 2.56-s epochs and analysed by FFT into conventional theta, alpha and beta 1 frequency bands. The EEG amplitude for each subject in each frequency range was averaged over a minimum of 60 s. Separate group averages for each sample were calculated and the resulting topographical distributions of electrical potential and current density were compared. The results indicated that the reproducibility of electrical potential in the theta and beta 1 frequency ranges was extremely poor and only approached acceptable levels in alpha. Reproducibility of current source density was poor in all frequency ranges. Although some improvement in reproducibility was obtained following spatial smoothing for alpha potential, the highest reproducibility achieved was only 0.65. Reasons for the poor reproducibility of topographical EEG and the implications of these findings are discussed. PMID- 9202999 TI - Foot and hand area mu rhythms. AB - Spontaneous EEG can display spatio-temporal patterns of desynchronized or synchronized alpha band activity. Event-related desynchronization (ERD) of rhythms within alpha and lower beta bands is characteristic of activated cortical areas ready to process information or to prepare a movement, while event-related synchronization (ERS) in the same frequency bands can be seen as an electrophysiological correlate of resting or idling cortical areas. EEG was investigated over primary sensorimotor and premotor areas during discrete hand and foot movements. ERD was found over the primary hand area during finger movement and over the primary foot area during toe movement. The former was observed in every subject, the latter was more difficult to find. From these results it can be speculated that each primary sensorimotor area has its own intrinsic rhythm, which becomes desynchronized when the corresponding area is activated. ERS, in the form of an enhanced mu rhythm on electrodes overlying the primary hand area, was observed not only during visual processing but also during foot movement. In both cases, the hand area is not needed to perform a task and, therefore, can be considered to be in an idling state. The supplementary motor area (SMA) also plays an important role in preparation and planning of movement. It is demonstrated that this area also displays rhythmic activity within the alpha band, that is both linearly and non-linearly phase coupled to the intrinsic (mu) rhythm of the primary hand area. With planning and preparation of movement, this SMA rhythm is desynchronized and also the degree of coupling between the two areas decreases. PMID- 9203000 TI - Driven and synchronized brain activities in the alpha band: a neuromagnetic test for frequency responsiveness. AB - A new procedure to study cortical rhythmical activity, which includes stimulation paradigm, data analysis, and data presentation, is proposed. It enhances several features of the rhythmical responses while allowing for very short measurement sessions. This procedure permits us to classify activities within the frequency window studied on the basis of their responsiveness to scanned rhythmical stimulation. It can be used to recognize functional and pathological indicators linked to such activities. When used in conjunction with the identification of functional correlates of the same activities, it may bring useful information for their modelling. PMID- 9203001 TI - A possible role of evoked alpha in primary sensory processing: common properties of cat intracranial recordings and human EEG and MEG. AB - Regarding the evoked potential (EP) as a superposition of evoked EEG rhythms in several frequency ranges, we investigated the following issue: Are distinct evoked rhythms, in particular the alpha (8-15 Hz) response, related to separable physiological processes? Frequency domain analysis of EPs was used to evaluate results of cross-modality experiments, i.e.: responses to auditory stimuli were simultaneously recorded from the auditory cortex (adequate stimulation) and from the visual cortex (inadequate stimulation). Responses to visual stimuli were recorded from the same sites. The results of these experiments and further measurements (EEG and MEG responses in humans, among them multiple sclerosis patients) are consistent in the following respect: The amplitudes of alpha responses are dependent on whether or not a stimulus applied is adequate. Alpha responses may thus be related mostly to primary sensory processing. In contrast, theta responses (4-7 Hz) were observed for adequate as well as inadequate stimuli. They may be related rather to associative and cognitive processing than to primary sensory processing. Thus frequency responses, in particular the alpha response, are not artificial results of digital filtering, but functionally significant brain responses. PMID- 9203002 TI - Phase correlation among rhythms present at different frequencies: spectral methods, application to microelectrode recordings from visual cortex and functional implications. AB - In classical EEG analysis rhythms with different frequencies occurring at separable regions and states of the brain are analysed. Rhythms in different frequency bands have often been assumed to be independent and their occurrence was interpreted as a sign of different functional operations. Independence has scarcely been proved because of conceptual and computational difficulties. It is, on the other hand, probable that different rhythmic brain processes are coupled because of the broad recurrent connectivity among brain structures. We, therefore, set out to find interactions among rhythmic signals at different frequencies. We were particularly interested in interactions between lower frequency bands and gamma-activities (30-90 Hz), because the latter have been analysed in our laboratory in great detail and had properties suggesting their involvement in perceptual feature linking. Fast oscillations occurred synchronized in a stimulus-specific way in the visual cortex of cat and monkey. Their presence was often accompanied by lower frequency components at considerable power. Such multiple spectral peaks are known from many cortical and subcortical structures. Despite their well known occurrence, coupling among different frequencies has not been established, apart from harmonic components. For the present investigation we extended existing analytical tools to detect non linear correlations among signal pairs at any frequency (including incommensurate ones). These methods were applied to multiple microelectrode recordings from visual cortical areas 17 and 18 of anesthetized cats and V1 of awake monkeys. In particular, we assessed non-linear correlations by means of higher order spectral analysis of multi-unit spike activities (MUA) and local slow wave field potentials (LFP, 1-120 Hz) recorded with microelectrodes. Non-linear correlations among signal components at different frequencies were investigated in the following steps. First, the frequency content of short (approximately 250 ms) sliding window signal epochs was analyzed for simultaneously occurring rhythms of significant power at different frequencies. This was done by a newly developed method derived from the trispectrum using separate averaging of the products of short-epoch power spectra for any possible combination of frequency pairs. Second, non-linear (quadratic) phase coupling between different frequencies was assessed by the methods of bispectrum and bicoherence. We found phase correlations at different frequencies in the visual cortex of the cat and monkey. These couplings were significant in about 60% of the investigated MUA and LFP recordings, including several cases of coupling among incommensurate (i.e. non harmonic) frequencies. Significant phase correlations were present: (1) within the gamma-frequency range; (2) between gamma- and low frequency ranges (1-30 Hz, including alpha- and beta-rhythms); and (3) within the low frequency range. Phase correlations depended, in most cases, on specific visual stimulation. We discuss the possible functional significance of phase correlations among high and low frequencies by including proposals from previous work about potential roles of single-frequency rhythms of the EEG. Our suggestions include: (1) visual feature linking across different temporal and spatial scales provided by coherent oscillations at high and low frequencies; (2) linking of visual cortical representations (high frequencies) to subcortical centers (low frequencies) like the thalamus and hippocampus; and (3) temporal segmentation of the sustained stream of incoming visual information into separate frames at different temporal resolutions in order to prevent perceptual smearing due to shifting retinal images. These proposals are, at present, merely speculative. However, they can, in principle, be proved by microelectrode recordings from trained behaving animals. PMID- 9203003 TI - Rhythms in the alpha band in cats and their behavioural correlates. AB - We describe two sets of rhythmical field potentials that may be recorded from the neocortex of the behaving cat: those at about 14 Hz ('mu' rhythms), in part of the somatic area SI; others at about 10 Hz ('alpha' rhythms), in part of the visual cortex (lateral gyrus). We report their precise localization, their thalamic correlates, their dependence upon a noradrenergic brainstem modulatory mechanism, and finally their possible functional significance. PMID- 9203004 TI - Low-frequency oscillations of visual, auditory and somatosensory cortical neurons evoked by sensory stimulation. AB - Low-frequency oscillations-LFOs-below 20 Hz in the activity of cortical neurons are a commonly observed property across all sensory modalities. However, the functional significance and potential role of these intrinsic oscillations are not well understood. Here, we attempt to provide a general framework for the interpretation of this phenomenon by considering its properties across several sensory modalities. In the first part, we provide a survey and a general description of low-frequency oscillations (LFOs) at a cellular level observed following adequate [Basar, and Schurmann, 1994]. Sensory stimulation of neurons recorded in three sensory modalities of neocortices in higher mammals. The second part will address some functional aspects of low-frequency oscillations (LFOs) such as stimulus selectivity and so-called 'interference' phenomena, specifically with findings related to 'resetting' and 'gating' of sensory processing streams. Finally, a hypotheses is outlined in which the low-frequency oscillations are regarded as an organizational principle by which continuity of sensory and motor states over time could be accomplished. PMID- 9203005 TI - The contribution of glial cells to spontaneous and evoked potentials. AB - The mechanism by which brain cells generate alpha and other rhythms remains obscure, and the possible participation of glial cells in the process continues to be debated. We will present data obtained from freely moving rats in which flashes produced by a light emitting diode implanted under the skin of the scalp evoke retinal and cortical responses recorded through electrodes implanted behind the eye and over visual cortex. In the retina, which is a brain-like structure isolated in the periphery during embryology, the b-wave evoked response is thought to be produced by the Muller glial cells as they maintain potassium ion homeostasis in the extracellular space during the synaptic events initiated by rod and cone activation. We will report on the results of a search in this retinal analogue of the brain for spontaneous activity in the EEG spectrum. PMID- 9203006 TI - Alpha rhythms: noise, dynamics and models. AB - Alpha rhythms appear as sinusoidal-like oscillations in the electroencephalogram (EEG) within the frequency range 8-12 Hz that waxe and wane in a more or less irregular way. The irregularity may have various origins. It may be due to noise or the oscillations may have an intrinsic irregular character, e.g. they may be generated by chaotic processes [Jansen (1991) Quantitative analysis of electroencephalograms: is there chaos in the future? Int. J. Biomed. Comput., 27: 95-123; Pradham, N. and Dutt, D.N. (1993) A nonlinear perspective in understanding the neurodynamics of EEG. Comput. Biol. Med., 23: 425-442; Pritchard et al. (1995) Dimensional analysis of resting human EEG II: Surrogate data testing indicates nonlinearily but not low-dimensional chaos. Psychophysiology. 32: 486]. The term noise is often used in neurophysiology with different connotations as pointed out by Bullock (1990), either meaning an unwanted signal from the point of view of the receiver of a message, or a signal with intrinsic random fluctuations, i.e. with a stochastic character. Here we consider noise in this sense, as random or quasi-random neural activity. In this overview, we concentrate on the question of whether alpha rhythms should be considered generated in neuronal networks (1) as forms of filtered noise, (2) as deterministic oscillations influenced by noise or (3) as the result of chaotic dynamics. A clear answer to this question can have theoretical value because it may lead to a general model of the generation of this important EEG signal. Such a model, of course, would be a macroscopic one, since it would primarily account for the properties of the alpha rhythms at the neuronal network level. A translation of these properties to the microscopic, i.e. neuronal, level will not be easy to achieve without more direct knowledge of the membrane and synaptic basic properties of the neurons involved. Here we consider the question formulated above by presenting some relevant experimental evidence and theoretical arguments. The consideration whether alpha rhythms may have noise or chaotic sources implies examining how and where such sources can occur in the neuronal networks of the brain. Therefore we present, first, some basic data regarding the possible origin of noise and of chaos in neuronal networks. Second, the signal analysis methods that have to be applied in order to discriminate between filtered noise activities and chaotic oscillations are introduced. Third, the implications of these signal analyses regarding the possible answer to the initial question are discussed. PMID- 9203007 TI - Non-linear dynamics of alpha and theta rhythm: correlation dimensions and Lyapunov exponents from healthy subject's spontaneous EEG. AB - The aim of the present paper was to analyze some non-linear dynamic properties of the resting EEG from healthy subjects under eyes closed conditions. For this purpose we digitally filtered the spontaneous EEG in the theta (3-8 Hz) and alpha frequency range (8-13 Hz) and considered these independent rhythms as signals from a deterministic system. Under certain conditions non-linear dynamic systems are able to generate deterministic chaos, which means that similar causes do not produce similar effects. This phenomenon is called sensitive dependence on initial conditions. From different lead positions (F3, F4, Cz, P3, P4, O1 and O2) we calculated the so-called correlation dimension D2, which is assumed to be an estimation of the system's complexity, and the Lyapunov exponent L, which appears to be a measure of the sensitive dependence on initial conditions. Our investigations revealed that the dimensionality of the theta- and alpha-rhythm varies within subjects across the experimental session in wide ranges. The degrees of freedom of the alpha and theta rhythms across the scalp are in the same order, indicating dynamic processes which can not be differentiated by applying the Grassberger-Procaccia algorithm. The Lyapunov-exponents, indicating 'how chaotic a deterministic process is', are in general smaller for the theta than for the alpha activity. Across the scalp there is no evidence for different dynamics of the theta rhythm. The dynamics of the alpha rhythm, on the contrary, appears to be different at various lead positions. It appears justified to state that the dynamics of the frontal alpha activity is functionally different from the alpha activity recorded at other topographic locations. PMID- 9203008 TI - Segregation of the thalamic alpha rhythms from cortical alpha activity using the Savit-Green S-statistic and estimated correlation dimension. AB - The large-amplitude, occipital alpha rhythm of the human electroencephalogram (EEG) is associated in normal subjects with a state of relaxed wakefulness. We analyzed resting eyes-open and eyes-closed human EEG data using three measures: power in the 8-12-Hz alpha frequency band, estimated correlation dimension (D2), and the Savit-Green S-statistic. At occipital loci, two groups of points were evident in the scatterplot of S vs. estimated D2. Group A was of higher dimension and consisted of predominantly eyes-open records. Group B was of lower dimension and consisted of more eyes-closed than eyes-open records. Furthermore, Group B had a broad range of alpha power, with alpha power being negatively correlated with estimated D2 (higher alpha power associated with lower dynamical complexity). In contrast, Group A had a very small range of low alpha power, and was positively correlated with estimated D2 (higher alpha power associated with increased dynamical complexity). Our results indicate that Group B EEG (the alpha rhythm) and Group A EEG (EEG containing 'other' alpha activity) have fundamentally different dynamical properties. PMID- 9203009 TI - EEG alpha dynamics as viewed from EEG dimension dynamics. AB - EEG alpha power covaries with changes in visual input and with changes in other aspects of cortical processing. We present an synopsis of three experiments that demonstrate these effects. A concurrent analysis of the EEG dimension indicates that the dynamics of EEG alpha may result from at least two different mechanisms. PMID- 9203010 TI - Visual and auditory evoked phase resetting of the alpha EEG. AB - An increasing number of studies over the last decade or so have suggested that evoked potential (EP) morphology is partially due to a reorganization of the phase of the ongoing EEG. Phase resetting is common to non-linear oscillatory systems in response to a perturbation and has been observed in a number of biological systems such as circadian rhythms and the ECG. However, it has not been studied adequately in the context of EP research. Five clinically normal male volunteers (age 25-32) were subjected to randomly occurring light flashes for approximately 1 h. EEG recordings were obtained from a midline parieto occipital site (POz) referenced to linked ears. An additional five male subjects (age 26-36) participated in an auditory P3 study of the effects of nimodipine, a calcium channel blocker, on brain electrical activity. Four of the latter subjects were diagnosed as crack cocaine abusers and one was negative for crack use. The single trials were bandpass filtered in the 8-13 Hz band and the phase angle at the moment of stimulation was computed. We examined the relationship between initial phase angle and both latency and amplitude of the first two post stimulus negative peaks. The results demonstrate that these peaks undergo phase and (pre-stimulus) amplitude sensitive latency reorganization during presentation of both visual flash stimuli and auditory non-target oddball stimuli in a P3 experiment. PMID- 9203011 TI - On the relationship between EEG and P300: individual differences, aging, and ultradian rhythms. AB - The literature on the relationship between background EEG and event-related brain potentials (ERPs) is reviewed, with the conclusion that variation in the former can contribute to individual variability in the latter. The effects of background EEG activity on the P300 component are then described using the results of three experiments. Study 1 assayed the association between EEG spectral power/mean frequency and P300 amplitude/latency measures in young adults. For the slower delta, theta, and alpha bands generally strong correlations who obtained for both types of measures. Study 2 employed similar techniques to assess a large sample of adults who varied in age from 20-80+ years. EEG power in the slower bands was correlated positively with P300 amplitude across the age range, but few effects for mean frequency/component latency were observed. Study 3 measured a group of young adults ten times very 20 min to assess for temporal changes in the relationship between EEG and ERPs. The correlations between spectral power and P300 amplitude measures were found to vary in a manner that suggested the influence of ultradian rhythms on neuroelectric activity. Taken together, the findings from all three study indicate that background EEG variation contributes significantly to the individual variability of the P300 ERP. Theoretical and applied implications of the findings are discussed. PMID- 9203012 TI - EEG-alpha rhythms and memory processes. AB - The results of several experiments indicate that alpha frequency varies as a function of memory performance. It was found that in samples of age matched subjects alpha frequency of good memory performers is about 1 Hz-higher than those of bad performers. The difference in alpha frequency between good and bad performers reaches a maximum during the retrieval of information, is much smaller during encoding and is minimal--but still significant--during a resting period. These results suggest that alpha frequency may be a permanent and not only a functional parameter that determines the speed with which information can be retrieved from memory. The calculation of changes in band power indicate further that the upper alpha band is particularly sensitive to semantic memory demands. The lower alpha band, on the other hand, seems to reflect attentional processes. These findings are discussed on the basis of a hypothesis which assumes that EEG frequencies within the alpha band stem at least in part from the thalamus and that the activity of thalamo-cortical networks reflects processes that are related to searching, accessing and retrieving information from (scmantic) long term memory. PMID- 9203013 TI - Some aspects of alpha activity in relation to a new generalized model for the EEG. PMID- 9203014 TI - A descriptive framework for information processing: an integrative approach. AB - Survival depends on veridical perception of the environment and appropriate response to it. How do high level organisms perceive and compare, in short, evaluate stimuli and how do they select and execute responses? How does information processing proceed? How does the mind work? PMID- 9203015 TI - Alpha rhythm parameters and short-term memory span. AB - The study examined a statement from the neurophysiological model of A.N. Lebedev as to whether short-term memory span is related to a ratio of the alpha-rhythm frequency and parameters of step-type behaviour of wave processes in the EEG alpha range. Forty young healthy subjects performed a digit memory test. EEG signals were recorded from the occipital area during a resting period with eyes closed. As one of the parameters of step-type behaviour of alpha oscillations a mean value of the difference between periods of neighbouring peaks in the alpha spectra was calculated. A second parameter--the duration of alpha spindles, considered to be a superposition of alpha oscillations--was also measured. It was shown that the ratio of the alpha-rhythm frequency to parameters of the step-type behaviour of alpha oscillations positively correlated with memory performance. PMID- 9203016 TI - Is there a way from behavior to non-linear brain dynamics? On quantal periods in cognition and the place of alpha in brain resonances. PMID- 9203017 TI - Normal and pathological changes in alpha rhythms. AB - Normal alpha rhythm begins to appear at three months. It was possible to observe successively, the rhythmicity, visual reactivity, occipital topography, and at the end of the first year, the first bursts of 7-8 Hz waves. Evolution of alpha was not linear with the age. Alpha frequency acceleration was important before the age of 11 years. In adults it decreased from 20 to 55, then there was a tendency to increase with aging. Amplitude was highest at 11-12 years, then its tendency was to decrease until 40, and to increase later. In pathology, for instance, alpha could be observed too early in newborn and infants with malformations, in children, in some psychiatric diseases, in adults and old people, alpha was replaced by theta waves in dementia: it seemed to be a very reliable sign, at the beginning of the disease. PMID- 9203018 TI - Alpha response system in children: changes with age. AB - Evoked and event-related brain potentials (Eps, ERPs) may be regarded as originating from the reorganization of the spontaneous EEG rhythms (Basar, 1980). Until now, no data is available about the development of the evoked frequency components in EPs and ERPs of children. The main objective of the present research was to study the alpha response system in 6-11-year-old children. We suggested that the ability to reorganize the alpha activity and produce repeatable alpha patterns after external stimulation might undergo developmental changes that could reflect certain changes in information processing with increasing age from childhood to adulthood. Fifty 6-11-year-old children divided into five age groups, and 10 young adults were studied in a passive and an odd ball condition. Alpha responses in the auditory EPs and non-target ERPs at Fz, Cz and Pz were analyzed. The magnitude and phase-locking with stimulus of single alpha responses were evaluated in the first 300 ms of the post-stimulus epoch. An original method was applied to assess quantitatively the repeatability (phase locking) of the evoked alpha oscillations. The magnitude and the phase-locking to stimulus were analyzed with respect to their dependence on the age and topography factors. Our main results show that the alpha responses in 6-11-year-old children are different from those in adults: (1) Adults had significantly lower amplitude and stronger phase-locking than children; (2) Adults had maximal alpha amplitudes and phase-locking over the vertex, whereas children displayed maximal responses over the parietal site; (3) The phase-locking of eldest (10-11-year-old) children was as strong as in adults. Whereas no difference existed between groups of children in alpha response amplitudes, a significant increase in phase-locking from 6 to 11 years was observed. Concerning the obtained results we suggest that (1) Alpha response system is functionally involved in 6-11-year-old children, though its development is not complete at the age of 11, the upper limit of our sample (2) With regard to their differential developmental time-courses, the magnitude and the phase-locking parameters might be suggested to relate to different functional aspects of the alpha response system. The applied original method makes it possible to analyze the phase-locking to stimulus (or phase reordering) separately and independently from the amplitude (enhancement) of the frequency responses, thus providing for a deeper examination of the evoked frequency components. PMID- 9203019 TI - Alpha EEG activity and subcortical pathology in HIV infection. AB - Quantitative electroencephalographic (QEEG) investigations of patients with HIV-1 infections were made to detect early signs of HIV brain involvement. QEEG data were recorded in a prospective controlled cohort study together with standardised clinical, immunological and neuropsychological tests. Subject groups were HIV seronegative controls, HIV-seropositive subjects with asymptomatic infection, and HIV-seropositive subjects with symptomatic infection exclusing AIDS defining illnesses. Marked increase in background alpha amplitude preceded cognitive and neurological impairment in the symptomatic stage of infection. Elevation of alpha amplitude was also associated with change in psychiatric status. Antiretroviral medication suppressed this alpha elevation, supporting the usefulness of QEEG in monitoring drug CNS effects. In the context of clinical, neuropathological and brain imaging data it is suggested that these changes in the alpha rhythm are the earliest, albeit unspecific, signs of HIV brain involvement, reflecting the predominantly subcortical prominence of the pathological process. PMID- 9203020 TI - The early history of EEG data-processing at the Massachusetts Institute of Technology and the Massachusetts General Hospital. PMID- 9203021 TI - Functional correlates of alphas panel discussion of the conference 'Alpha Processes in the Brain'. PMID- 9203022 TI - The place of Behcet's syndrome among the autoimmune diseases. AB - The most popular research interest in Behcet's Syndrome (BS) is directed to immunological mechanisms. However there are many ways in which BS differs from a classic autoimmune disease. The most important differences lie in the male dominance in severe disease, lack of association with other autoimmune diseases, lack of association with HLA alleles usually seen in autoimmune diseases, lack or paucity of autoantibodies and B cell hyperfunction-especially Sjogren's syndrome- and no definite T cell hypofunction in BS. Perhaps less important points are the peculiar geographic and ethnic distribution, the peculiar clinical features and the lack of response of BS to steroids. The value of immunological data on BS will much increase of we include in each experiment patients with classic autoimmune diseases along with "pure" inflammatory conditions like gout and infectious diseases. PMID- 9203023 TI - Hyperreactivity of neutrophils and abnormal T cell homeostasis: a new insight for pathogenesis of Behcet's disease. AB - Neutrophil hyperfunction of Behcet's disease is in part regulated by genetical factors, especially related to HLA-B51 genes and by immunological abnormalities. Regarding the latter points, Behcet's disease worsens with abnormal regulation by gamma delta T cells and alpha beta T cells. Indeed, our own studies and those of other laboratories suggest that human heat shock protein may be one of the triggering factors for activation of both the gamma delta T cells and alpha beta T cells. These activated T cells may produce proinflammatory and/or inflammatory cytokines, and lead to tissue injury possibly via delayed-type hypersensitivity reaction, macrophage activation, and activation and/or recruitment of neutrophils. Here we discuss the crosstalk between the immune system and neutrophils in this disease. PMID- 9203024 TI - The role of heat shock protein, microbial and autoimmune agents in the aetiology of Behcet's disease. AB - Investigation of the aetiology of Behcet's disease (BD) has focused predominantly on herpes simplex virus immunopathology, autoimmunity to oral mucosa or cross reactive microbial antigens, and streptococcal infection. These aetiological factors might have a common denominator in microbial heat shock protein (HSP) which shows significant homology with the human mitochondrial HSP. Indeed, the uncommon serotypes of Streptococcus sanguis found in BD cross-react with the 65 kD HSP which also shares antigenicity with an oral mucosal antigen. T cell epitope mapping has identified 4 peptides derived from the sequence of the 65 kD HSP which stimulate specifically TCR gamma delta + lymphocytes from patients with BD. These peptides (111-125, 154-172, 219-233 and 311-325) show significant homology with the corresponding peptides derived from the human 60 kD HSP. The specific proliferative response of TCR gamma delta + lymphocytes elicited by the 4 peptides can be used as a laboratory test for the diagnosis of BD. The pathogenic significance of these peptides has been established by inducing uveitis in rats. PMID- 9203025 TI - Pathogenic gene responsible for the predisposition of Behcet's disease. AB - HLA-B51 is well known to be associated with Behcet's disease (BD) in many different ethnic groups. The hypothesis may be presented that B51 molecules are primarily involved in BD development through specific antigen presentation. Furthermore, HLA-C genotyping by the polymerase chain reaction-sequence specific primers method suggests that the BD pathogenic gene is not the HLA-C gene itself but some other gene located near the HLA-B gene. Polymorphic analysis of the Tau a microsatellite between the HLA-B and TNF genes indicates that the pathogenic gene of BD is not the HLA-B51 gene itself but other gene located around the HLA-B gene. Recent studies suggest that many novel genes exist in the region between the TNF and HLA-B or HLA-C genes such as MIC and PERB, etc. and furthermore, many unidentified new genes have been suggested to exist in this region. In this paper, the present situation of the investigations on the genetic predisposition responsible for BD was reviewed. PMID- 9203026 TI - Immunotherapy for Behcet's disease. AB - Behcet's disease is an inflammatory disorder affecting many organs including the eye and one of the most common sight-threatening causes in countries around the Mediterranean basin and in the Asia including Japan, Korea and China. A number of clinical and laboratory findings suggest the significant involvement of the immune alterations in the pathophysiology and the pathogenic mechanisms of Behcet's disease. The immune alterations demonstrated in the disease include the alteration of the T cell circuitry and abnormal functions of the leukocyte. Because immunologic processes are believed to be in the chain of events in the manifestation of Behcet's disease, various agents capable of modulating the immune responses have been used to treat the disease. These drugs include corticosteroids, colchicine, cytotoxic agents, and immunophilin ligands (cyclosporine and FK506). This paper reviews the experimental and clinical investigations to analyze the immunopharmacological activities of these immunosuppressive agents in animal models and in patients with Behcet's disease. PMID- 9203027 TI - Uveitis in Behcet's disease. AB - The ocular complications of Behcet's disease are considered one of the major criteria upon which the diagnosis is based. Its complications are frequently sight threatening and require constant attention. The ocular disease is characterized by repeated, explosive ocular inflammatory attacks which get better by themselves, so that in-between attacks there is little or no evidence of inflammatory disease in the eye. The anterior segment can be involved alone, most frequently presenting as a severe anterior uveitis, frequently with hypopyon. This is not associated with a poor visual outcome, and usually treated with topical medication to make the patient more comfortable. However, the anterior segment disease is usually accompanied by recurrent retinal vaso-occlusive disease which is sight threatening if repeated attacks occur. Treatment is with systemic medications, including corticosteroids, cyclosporine, FK506, anti metabolites, and cytotoxic agents. Complications of the inflammation can include retinal and optic atrophy, vitreous hemorrhage, neovascular glaucoma, and retinal detachment. PMID- 9203028 TI - Vasculitis in Behcet's disease. AB - Vasculitis underlies most of the characteristic lesions of Behcet's disease and is obligatory for the diagnosis in populations at risk. Sporadic cases and similar presentations in non-Silk-Route patients should be labelled as Behcet's syndrome and carry different prognosis because of differences in underlying pathologic changes and pathogenesis. Ethnicity and vasculitis should be included in diagnostic criteria. Large vessel involvement is characteristic if less common. PMID- 9203029 TI - New perspective on Behcet's disease. AB - There are many distinct differences between Behcet's disease of Silk Route and that of outside Silk Route; genetic factors, role of neutrophils, and severity of this disease. We have thus emphasized that we prefer the term "Behcet's syndrome" rather than "Behcet's disease". In this chapter, Behcet's disease seen along the Silk Route will be mainly discussed. HLA-B51 molecules themselves may be responsible, at least in part, for the neutrophil hyperfunction in Behcet's disease; a significant correlation was observed between the neutrophil hyperfunction and the possession of HLA-B51 phenotype, regardless of the presence of the disease, in both humans and HLA-B transgenic mice. T cells in this disease, proliferated vigorously in response to a specific peptide of human heat shock protein (HSP)-60; however, T cells from normal subjects or patients with rheumatoid arthritis, did not. This peptide has the amino acid sequence of 336 351 of human HSP-60, which is similar, but not identical to specific peptide of mycobacterial HSP-65. We have further analyzed T cell receptor (TCR) usage of HSP responsive T cells by means of TCR V beta subfamily specific monoclonal antibodies and polymerase chain reaction and single strand conformation polymorphism-based technique. We found that T cells with specific TCR V beta subfamilies proliferated and increased in number in response to the peptide by an antigen-specific fashion. The result of recurrent exposure to the HSP may break the tolerance to self-HSP, and provoke T cell responses to self- and microbial HSP. Such T cells produced Th1-like proinflammatory and/or inflammatory cytokines. This leads to tissue injury, possibly via delayed-type hypersensitivity reaction, macrophage activation, and activation and/or recruitment of neutrophils. Our data shed a new light on the autoimmune nature of Behcet's disease; a novel multistep molecular mimicry mechanisms may induce and/or exacerbate Behcet's disease by bacterial antigens that activate T cells previously educated by self-peptides of HSP. This would lead to positive selection of autoreactive T cells in this disease. PMID- 9203030 TI - A case of pure renal hydatid cyst without multiple organ involvement. AB - Hydatid disease of the urinary tract is very rare. We report a case of pure renal hydatid cyst, with its clinical presentation and management. PMID- 9203031 TI - Activities of superoxide dismutase and glutathione peroxidase enzymes in cancerous and non-cancerous human kidney tissues. AB - The activities of superoxide dismutases (total, cytoplasmic and mitochondrial) and glutathione peroxidase were measured in 10 cancerous and 10 non-cancerous adjacent human kidney tissues. Total (T-SOD) and cytoplasmic (Cu, Zn-SOD) superoxide dismutase and glutathione peroxidase (GSH-Px) activities were found lower in cancerous tissues compared with those of non-cancerous ones. However, no difference was found between the mitochondrial (Mn-SOD) superoxide dismutase activities of the tissues. Similarly, no differences were observed between the enzyme activity values of the tissues at stage I-II and III-IV renal cancer. In correlation analysis the positive relation found between Cu, Zn-SOD and GSH-Px enzymes in the non-cancerous tissues was found to be absent in the cancerous ones. The results suggest that enzymatic free radical defense mechanism is significantly reduced in the cancerous human kidney tissues due to depressed Cu, Zn-SOD and GSH-Px activities. PMID- 9203032 TI - Iatrogenic injuries to ureter, bladder and urethra during abdominal and pelvic operations. AB - Abdominal and pelvic operations at Departments of Obstetrics and Gynaecology, and General Surgery play an important role in ureteral, bladder and rarely urethral injuries. Fifty-nine patients with iatrogenic ureteral, bladder and urethral injuries were treated at the Department of Urology, Ataturk University Research Hospital, between 1985 and 1995. These injuries were urinary vaginal fistulas in 43 patients (vesicovaginal 33, ureterovaginal 7, urethrovaginal 2 and vesicovaginal plus urethrovaginal 1), ureteric ligation in 9, bladder laceration in 7. These injuries were treated by different methods. All patients were followed up by intravenous urography (IVU) and urine culture three months later. It must be borne in mind that iatrogenic urinary tract injuries are not rare. Bladder and ureteral catheterization must be performed to prevent these complications. PMID- 9203033 TI - Multiple metastases of a malignant cutaneous melanoma in the cavitary system of the upper urinary tract. AB - Multiple metastases of malignant cutaneous melanomas in the upper urinary system are encountered very rarely. Such a case, treated surgically by the authors, is presented. A short review of similar cases in the literature is also given. PMID- 9203034 TI - Correlation of nuclear p53 over-expression with clinical and histopathological features of transitional cell bladder cancer. AB - Forty-four pathologic specimens of 39 bladder cancer patients were analyzed immunohistochemically with D07 monoclonal antibody to detect over-expression of mutant p53 gene. The findings were interpreted by correlating with patient age, sex, cigarette smoking, number and macroscopic appearance of tumour, histological tumour grade, muscular invasion, vascular invasion, necrosis and urothelial atypia or dysplasia. Mutant p53 gene was over-expressed in 8 (18.2%) specimens. Statistically significant correlation with grade, vascular invasion, necrosis and patient sex was found with p53 over-expression. Available follow-up data were insufficient to draw a conclusion about the prognostic role of p53 over expression. Prospective studies with larger number of patients are needed to define the exact place of nuclear p53 over-expression in transitional cell bladder cancer. PMID- 9203035 TI - Massive haemorrhage of inoperable bladder carcinomas: treatment by intravesical formalin solution. AB - Numerous modalities of treatment have been used in the past to control massive bladder haematuria, with varying degrees of success. Formalin has been used in urology only for the treatment of intractable haematuria of inoperable bladder carcinomas, usually as the last resort when all other nonsurgical attempts have failed and before more aggressive surgical measures are considered. Eight patients with bladder tumours classified T2 (2 cases), T3 (2 cases) or T4 (4 cases) and 2 patients with radiation cystitis were assessed as being beyond the scope of even palliative surgery, severe haemorrhage being present in all cases. The treatment was instituted in all cases by intravesical instillation of a 10 per cent formalin solution under general anaesthesia. Four patients received 4 and 6 instillations, respectively, the former over 4 weeks and the latter over 10 months. The bladder was filled completely and an indwelling-catheter introduced, the formalin solution being left in the bladder for 5 to 30 min (mean: 12 min). Haematuria was absent after 1 to 25 days (mean: 11 days) in 9 cases. The 10th patient died before arrest of haemorrhage. Survival after instillation was 65 days to 27 months (mean: 11.5 months). The outcome was fatal within 4 months or less in 3 cases and 4 patients died of renal failure within 3 months, one within 65 days after instillation. In 4 cases, treatment with formalin reduced bladder capacity to less than 100 ml. Other complications included retroperitoneal fibrosis (1 case), urinary incontinence (3 cases) and severe frequency and nocturia (3 cases). This procedure should therefore be reserved for terminal cases unable to support more aggressive therapy. PMID- 9203036 TI - Bladder replacement with modified Studer pouch using absorbable staples. AB - The use of absorbable staples may allow to reduce the operating time of orthotopic bladder replacement. We report our experience in 14 patients with a modification of the Studer technique using absorbable PolyGIA instruments. The technique has been shown to be simple and quick to perform (time for pouch creation 15-40 minutes, mean 16 minutes) with no significant intraoperative difficulties. Urodynamic data and continence are satisfactory and seem comparable to different procedures and not related to staples' use. The main question remains if the reduced operating time equalizes the high cost of staple devices. PMID- 9203037 TI - Urine leakage from the umbilicus in a child with achondroplasia and tetraplegia (due to cervical stenosis): a safety vent for the obstructed neuropathic bladder. AB - Urine leakage from the umbilicus was observed while expressing urine by the Crede manoeuvre in a three-year-old tetraplegic girl with a chronically distended urinary bladder. Intravenous urography (IVU) revealed bilateral hydroureteronephrosis with markedly distended urinary bladder. Regular three hourly intermittent catheterization was advised, and the parents and carers of this child agreed to perform catheterization. There was cessation of urine leak within 48 hours of urethral drainage. Cystography performed two weeks later showed no vesicoureteric reflux; vesicoumbilical fistula was no longer demonstrable. Follow-up IVU, performed after eight and half months of regular intermittent catheterization, showed regression of hydroureteronephrosis. We believe that urine leakage from the umbilicus served two important protective functions in this child, viz. (1) it prevented possible vesical or renal rupture; (2) the striking clinical symptom of urine leak from the umbilicus focussed the attention of the carers to the underlying serious condition of the urinary tract. Further, this case demonstrates that regression of marked hydroureteronephrosis can be achieved by intermittent catheterization performed at regular intervals by devoted parents/carers, in selected cases of spinal cord injury with neuropathic bladder, and vesical outlet obstruction, thus obviating the need for any form of temporary or permanent urinary diversion. PMID- 9203038 TI - Treatment with neodymium: YAG laser in patients with chronic prostatitis: a preliminary report. AB - A group of 30 patients with chronic abacterial prostatitis or prostatodynia, all of whom were unresponsive to conventional treatments, underwent transurethral neodymium: YAG laser therapy. The patients were evaluated with both objective and subjective parameters before and after treatment. Following treatment, the preliminary results at six months showed positive response (complete plus partial) rates in subjective parameters ranging from 47% to 86%. On the other hand, objective parameters including mean international prostate symptom scores (IPSS), mean quality of life indexes (QoL), mean uroflowmetric measurements and mean leukocyte count in expressed prostatic secretions showed marked improvements which were meaningful statistically. In 4 of 7 patients who underwent needle biopsy of the prostate, histology revealed definite inflammatory infiltration changes. Control biopsies following treatment showed almost complete disappearance of these changes. These preliminary results open up a new treatment modality in the management of this condition which has so far responded poorly to conventional therapy. PMID- 9203039 TI - Bladder mucosa in posterior urethral repair. AB - The case of a 10-year-old male child with posterior urethral stricture due to infections caused by indwelling urethral catheters treated by patch urethroplasty using free bladder mucosal graft is described. PMID- 9203040 TI - Long-term results of microsurgical drainage for idiopathic varicocele. AB - The long-term results of microsurgical shunts for idiopathic varicocele are reported in the present paper. Sixty-two patients with a total of 65 varicoceles (three were bilateral) were followed up for 1 to 8 years. Pre- and postoperative ultrasonographic evaluation of varicocele size was considered of great importance in order to reduce the bias of subjective clinical diagnosis and to achieve a reliable and objective follow-up. Microsurgical shunts were tailored to the type of reflux: renospermatic (76.9%), iliospermatic (10.8%) or mixed type (12.3%), 94% of patients experienced a complete morphologic disappearance of varicosities, while in 6% of the cases a consistent reduction of size was objectified although varicosities were still detectable at ultrasonographic examination. In patients with severe infertility a significant increase of seminal parameters was observed postoperatively and this improvement showed a higher statistical significance in patients aged < 30 years. PMID- 9203041 TI - Comparison of inguinal and laparoscopic approaches in the treatment of varicocele. AB - To determine the pros and cons of inguinal and laparoscopic varix ligation techniques, we reviewed 53 patients who underwent inguinal (n = 35) and laparoscopic (n = 18) varicocelectomy at two centers. Intraoperative complications were not observed in either of the groups. There was 1 recurrence and 1 persistence in the laparoscopically treated patients. The inguinal approach had the advantage of shorter operating time (19.1 versus 52.8 min), ability to ligate the external spermatic veins, and it could be performed as an outpatient procedure. However, the laparoscopic approach seemed superior for preserving the spermatic artery (88.8% versus 68.5%) and had lesser postoperative morbidity. PMID- 9203042 TI - In vitro drug sensitivity testing of human testicular germ cell tumours with cytostatic drugs and interferon alpha-2b. AB - In spite of the significant advances in the chemotherapy of germ cell neoplasms, some patients do not achieve disease-free status and ultimately die from their diseases. Therefore, it is reasonable to select the best chemotherapeutic agents in these patients by in vitro drug sensitivity assay (IVDSA) in order to apply the most effective agent in case of resistance to primary chemotherapy. Fresh operative cells from 12 testicular germ cell tumours (TGCT) were cultured in vitro. Sensitivity of the tumour cells to interferon-alpha (IFN-alpha), cisplatin, mitomycin C, vinblastine, doxorubicin, etoposide, bleomycin, vincristine (VCR) were tested by a colorimetric assay using MTT. A preexposure viability over 75% was essential for IVDSA. Sensitivity was determined by a more than 50 +/- 2 SD% reduction from the control absorbance. All eight drugs in their high concentrations exhibited cell proliferation inhibition in 83.3 +/- 100% of TGCT. But in low concentrations efficacy of IFN and VCR were found to be lower than the others (33.3% and 58.3%, respectively). The results indicated that although TGCT are highly sensitive to various agents IVDSA may help to identify the effective agents which might be necessary for second line chemotherapy in a small percentage of patients. PMID- 9203043 TI - Penile fractures and our treatment policy. AB - To evaluate our diagnosis and treatment policy of penile fractures, we reviewed 13 cases treated between August 1992 and August 1995. In ten patients early surgical correction, in one patient an elective surgical intervention was carried out. Two patients with minimal haematoma were suggested to undergo conservative treatment, but only one of them came to the control examination and he complained of fixed induration, pain and mild deviation in erection. In the group of early surgical repair there was no complication. In a patient who had had a delayed procedure small induration and minimal deviation were found. In the light of these findings we think that in penile fractures early surgical repair produces the best results. PMID- 9203044 TI - What is the role of lipoprotein abnormalities and platelet aggregation defects in cardiovascular disease in chronic dialysis patients? PMID- 9203045 TI - Fluvastatin (Lescol) treatment of hyperlipidaemia in patients with renal transplants. AB - Hyperlipidaemia of 18 male and 20 female patients following successful renal transplantation was treated with daily 20 mg fluvastatin (Lescol) for 12 weeks. The patients were several months after transplantation, and their total cholesterol levels exceeded 6.5 mmol/l following an 8-week diet. The effect of fluvastatin on the levels of total cholesterol, HDL, LDL, triglyceride, Apo A1 and Apo B, as well as of lipoprotein(a) was examined. Furthermore, changes of the renal function (GFR-urea, creatinine, uric acid) and hepatic function (bilirubin, GOT, GPT, CPK, ALP) were followed up, together with the body weight and blood pressure. The results of the examinations are summarized as follows: Fluvastatin may be administered effectively and without side effects in a daily dose of 20 mg in appropriately selected renal transplant patients. The average total cholesterol values, which were 7.91 mmol/l in men and 7.78 mmol/l in women following the diet, were reduced by 22-25% (p < 0.001) after 6 and 12 weeks, respectively, of fluvastatin treatment. The levels of LDL also decreased significantly (p < 0.001): in response to a 20 mg evening dosage, reduction of more than 25% was observed in 78% of men and 65% of women. Reductions of the Apo B levels were more pronounced in the females (18.3% men vs. 21.2% women). The ratio C/HDL-C decreased both in men (from 5.49 to 4.19) and in women (from 4.83 to 4.02). The ratio Apo B/Apo A1 also decreased (men: from 0.86 to 0.73, women: from 0.73 to 0.66). The concentrations of HDL and Apo A1 did not increase significantly, the reductions in the levels of triglyceride and lipoprotein(a) were not considerable either. An increase in the levels of hepatic enzymes and CPK was not encountered during the administration of fluvastatin. In two patients the levels of serum bilirubin increased by 2-4 micromol/l. Three patients complained about temporary myalgias of the sacroiliac or lumbar region which, however, were not accompanied by elevated CPK levels. The monitored levels of cyclosporine, urea and creatinine did not increase significantly during the 12 weeks of treatment. Two patients had temporary gastric complaints. PMID- 9203046 TI - Continuous ambulatory peritoneal dialysis. Experience in a single renal unit in the UK. AB - The outcome of 106 patients started on continuous ambulatory peritoneal dialysis (CAPD) in a single renal unit over a 3-year period was reviewed in order to compare our experience with that of other centres. The incidence of peritonitis was not different in diabetics compared with non-diabetics, in the elderly compared with younger patients, and in non-compliant compared with compliant patients. The patients' level of education had a significant effect on the number of days required to train for CAPD. Diabetics and non-compliant patients required more hospitalization. The need for community support increased the incidence of peritonitis. PMID- 9203047 TI - Plasma erythropoietin level and iron reserves in haemodialysis patients with and without acquired cystic kidney disease. AB - Concentration of erythropoietin (Epo) and iron reserves (IR) belong to the essential factors determining erythropoiesis in haemodialysis patients. Patients on dialysis with acquired kidney disease (ACKD+) can control anaemia better than patients without acquired kidney disease (ACKD-). Therefore we decided to check if plasma Epo levels and IR differ significantly in both groups of patients. Forty chronically haemodialyzed patients after ultrasound diagnosis were divided into 18 patients (45%) with ACKD+ and 22 (55%) without ACKD-. In both groups of patients we compared their plasma levels of Epo and IR. Plasma erythropoietin and ferritin levels were measured by enzymatic immunoassay. Iron reserves were estimated by the formula: IR = 400 x [ln (ferritin)-ln (50)]. In the ACKD+ group 72% of patients and in the ACKD- group 32% of patients did not require rHu Epo therapy. Plasma levels of erythropoietin and iron reserves did not differ significantly between ACKD+ and ACKD- patients. There must be also other factors than erythropoietin levels and iron reserves regulating erythropoiesis in these patients. PMID- 9203048 TI - The effect of calcitriol on renal anaemia in patients undergoing long-term dialysis. AB - In long-term haemodialysis patients suffering from secondary hyperparathyroidism Se iPTH could be suppressed by an intravenous calcitriol therapy. As the Se iPTH level became reduced, lower doses of Rh-EPO were already sufficient for the maintenance of the target haematocrit, while in two patients not requiring Rh-EPO treatment an improvement of their moderate anaemia could be observed. These data corroborate the view that calcitriol is an effective drug in secondary hyperparathyroidism. The results suggest that in the improvement of renal anaemia of dialysed patients the reduction of the Se-PTH level also plays a role. PMID- 9203049 TI - Transgenic animal technology. AB - There are numerous tools available to modify the genetic makeup of animals. They are being used to good advantage for studying basic biological phenomena. Within the decade, biomedical products derived from transgenic animals will be available, but the use of this technology for enhancing the quality and efficiency of livestock production will await further refinements in the technology. PMID- 9203050 TI - Progestin-androgen combination regimens for male contraception. PMID- 9203051 TI - Evidence for a role of intracellular-calcium release in nitric oxide-stimulated relaxation of the rabbit corpus cavernosum. AB - Erection is mediated by relaxation of the smooth muscle elements within the sinusoids of the corpus cavernosum. Although cavernosal relaxation can be mediated by a variety of mechanisms including purinergic stimulation, prostoglandins, and beta-adrenergic stimulation the major mechanism involves the stimulated release of nitric oxide (NO) and subsequent relaxation of the corporal smooth muscle. Experimentally, NO can be released both by direct stimulation of NO-containing nerves (using field stimulation) and indirectly via cholinergic stimulation of NO release from the endothelium (using bethanechol). Preliminary studies have indicated that NO release and/or NO-stimulated relaxation of corporal smooth muscle is an active process involving both an increase in cytosolic calcium and an increase in metabolic energy utilization. Ryanodine is a pharmacological agent that can inhibit calcium-stimulated calcium release from the sarcoplasmic reticulum. The results of the current study demonstrated that ryanodine inhibited both field-stimulated relaxation and bethanechol-stimulated relaxation but did not affect relaxation induced by adenosine triphosphate (ATP) or nitroprusside. These studies strongly support the hypothesis that NO stimulated relaxation is mediated, in part, by calcium release from the sarcoplasmic reticulum through ryanodine-sensitive channels. PMID- 9203052 TI - The effects of spinal cord injury on the status of messenger ribonucleic acid for TRPM 2 and androgen receptor in the prostate of the rat. AB - The prostate is one of the male accessory sex glands that produce fluid components of the seminal plasma. In addition to androgen, a normal innervation of the prostate is believed to be important for maintaining normal function of the prostate. Previously we noted that, in the rat, the weight of the prostate decreased following surgically induced spinal cord injury (SCI). This observation suggests that growth, and possibly function, of the prostate may be compromised after SCI. To explore this possibility, we examined the effects of SCI on the androgen-related biochemical properties and morphology of the prostate in the rat at various times after surgically induced SCI. SCI resulted in an acute decrease in prostate weight and an increase in steady state level of mRNA for testosterone repressed prostate message 2 (TRPM 2) during the first 2 weeks postinjury. These changes perhaps relate to an increase in cell death or a decrease in secretory activity due to an acute suppression of serum testosterone after the injury. Concomitantly, there was a transient, but significant, decrease in the steady state level of androgen receptor (AR) mRNA in the prostate during the first 2 weeks after SCI, an indication of an altered autoregulation of AR by its own ligand. Despite the fact that growth of the prostate, as indicated by weight increase, in SCI rats resumed 2 weeks postinjury, prostate weights were persistently lower in SCI rats than sham-operated controls for at least 3 months. Furthermore, prostate TRPM 2 mRNA levels remained elevated throughout the recovery period even after a normal prostate weight had been restored. In addition, a decrease in the height of ventral prostate epithelial cells was noted in SCI rats 28 and 90 days postinjury. These results demonstrate a prolonged effect of SCI on prostate function. These findings and our unreported observation of persistently smaller seminal vesicles in the same groups of SCI rats suggest that functions of male accessory sex glands may also be compromised after SCI. These changes may affect biochemical properties of the secretory products of these glands and may provide some explanation for the reported changes in the composition of the seminal plasma and abnormal sperm motility seen in the semen of SCI men. PMID- 9203053 TI - Expression of clusterin/sulfated glycoprotein-2 under conditions of heat stress in rat Sertoli cells and a mouse Sertoli cell line. AB - Clusterin is the major protein produced by rat Sertoli cells and is deposited onto sperm membranes; however, its function is unknown. In order to gain insight into the regulation of clusterin in Sertoli cells, the objective of the present study was to develop a model where the expression of clusterin could be affected in Sertoli cells in vitro. Rat Sertoli cells and mouse Sertoli cells (MSC1) were cultured under heat stress conditions (41 degrees C) for up to 48 hours. The mRNA for clusterin in Sertoli cells was compared to that in human epitheliod cancer cells (A431) to determine if clusterin expression was regulated in a lestis specific manner. The mRNA for heat shock protein 70 (HSP70) was also examined as it is a known stress-regulated gene. Expression of HSP70 mRNA was increased in all three cell types by 4 hours after the start of heat stress. Clusterin mRNA was increased over that of controls by 4 hours in heat-stressed A431 cells but did not significantly increase in MSC1 or Sertoli cells until 12 hours (P < 0.05). The induction of clusterin mRNA in MSC1 cells continued for at least 48 hours and required the sustained exposure of cells to the 41 degrees C temperature. The increase in the amount of clusterin mRNA was not due to an increase in transcript half-life, as determined by the addition of actinomycin D to the media of control vs. heat-stressed MSC1 cells. From the development of this in vitro model, we have seen that the timing of induction of clusterin by heat stress is Sertoli cell specific and is different than that of HSP70. This response in surviving cells during heat stress may be protective in that clusterin would bind to toxic compounds or solubilize cellular debris released by degenerating cells. PMID- 9203054 TI - Sertoli cells in culture and mRNA differential display provide a sensitive early warning assay system to detect changes induced by xenobiotics. AB - We have used cultured rat Sertoli cells as an "early warning system" to monitor for morphological and biochemical changes induced by two different xenobiotics cadmium acetate and polychlorinated biphenyls (PCBs). Sertoli cells begin to round, become vacuolized, and detach from their substrate within 24 hours of culture in the presence of cadmium at concentrations of 0.5-1.0 microM. Similar results were obtained with a lower dose of cadmium (0.01 microM) after 72 hours. When Sertoli cells are cultured for 24 hours in the presence of a mixture of PCBs (3,3',4,4'-tetrachlorobiphenyl, 2,2',4,6,6'-pentachlorophenyl, and 2,2',3,3',4,5,5', 6,6'-nonachlorobiphenyl) at concentrations of 1.0-2.0 microM, they enlarge. After 72 hours, a lower dose of PCBs (0.01 microM) produces similar cellular enlargement. Despite their changes in morphology, no reduction in Sertoli cell viability was seen at any of the concentrations or time points studied for either toxicant. Using mRNA differential display, a number of novel cDNAs were detected when cells were cultured with either cadmium or the PCBs, demonstrating that changes in gene expression accompany the changes in Sertoli cell structure. We propose that Sertoli cells in culture and mRNA differential display provide a sensitive morphological and biochemical assay system to detect early direct effects of low concentrations of toxicants on mammalian Sertoli cells. PMID- 9203055 TI - Leydig cell apoptosis in response to ethane dimethanesulphonate after both in vivo and in vitro treatment. AB - The biological effects of ethane dimethanesulphonate (EDS) are unique since cytotoxicity in the adult rat is almost exclusively confined to the Leydig cells. For this reason, EDS has been used extensively to investigate the physiological role of the Leydig cell and its products. Experiments were conducted to determine whether the Leydig cell will undergo apoptosis in response to EDS or methylprednisolone (MP), a glucocorticoid known to cause apoptosis in a number of other cell types. Percoll-purified Leydig cells were incubated for 24 hours with EDS (750 micrograms/ml), at which time the cells attached to the culture plate became rounded up while control cells were flattened and polyhedral. Following incubation with EDS or MP (10 microM), cells that became detached from the plate were characteristically apoptotic when stained with the fluorescent DNA dye, acridine orange. These cells had shrunk and the nuclear chromatin had become condensed, which is an early characteristic of apoptosis in other cells; eventually, apoptotic bodies formed, reflecting a later apoptotic stage. Electrophoresis of DNA extracted from the treated Leydig cells exhibited the characteristic ladder of the apoptotic process. Increasing the concentration of EDS or MP resulted in a dose-dependent increase in the incidence of apoptosis that reached a maximum of 25% (EDS) or 12% (MP) of detached cells. Administration of EDS in vivo caused a 20-fold increase in the number of apoptotic cells observed in interstitial cell preparations. In conclusion, the data indicates that programmed cell death, apoptosis, can occur in the Leydig cell and that this is the likely mechanism by which EDS kills the cells in vivo and in vitro. PMID- 9203056 TI - Evidence of proacrosin molecule abnormality as a possible cause of low acrosin activity and unexplained failure of fertilization in vitro. AB - The aim of this study was to investigate whether the molecular abnormality of proacrosin is associated with an unexplained failure of in vitro fertilization (IVF) and low acrosin activity in human infertility. Seventeen infertile men, whose spermatozoa appeared to be normal but did not fertilize using a conventional IVF technique, were included in this study. Proacrosin molecules from each patient were screened with electrophoresis and Western blotting using either anti-human proacrosin or anti-boar acrosin polyclonal antibodies. The bands of proacrosin appeared equally on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) at an identical site and reacted equally with human proacrosin antibodies in all patients studied. The anti-boar acrosin antibodies cross-reacted with the proacrosin bands of the patients in all but two cases. The two patients also demonstrated lower levels of total acrosin activity and different peptide maps of the isolated proacrosin. In conclusion, the molecular abnormality of proacrosin, which is reflected by a lack of cross reactivity to the anti-boar acrosin antibodies, is associated with low acrosin activity and may explain some cases of unexplained failure of human IVF treatment. PMID- 9203057 TI - Use of a rat cDNA probe specific for the Y chromosome to detect male-derived cells. AB - A cDNA probe that exhibits specificity for the rat Y chromosome was generated by using a set of primers specific to the murine Sry gene, the sex-determining region of the Y chromosome. A 459-base pair (bp) DNA fragment was obtained by polymerase chain reaction (PCR) amplification from male, but not female, rat genomic DNA (EMBL Nucleotide Sequence Database accession number X89730). This DNA fragment was purified, cloned using a vector, and digested with EcoR1 to yield a 270-bp DNA fragment. This 270-bp cDNA fragment, when used as a probe in Southern blot analysis of rat DNA, was observed to bind to three separate bands of approximately 2.3, 5.0, and 7.0 kb in size. The binding was demonstrated with male, but not female, genomic DNA. Another set of primers was generated to sequences within the 270-bp fragment that produced a PCR product of 104 bp. This DNA fragment, when used as a probe in Southern blot analysis, enabled PCR detection of at least 0.1% male cells in a mixed population of female cells. These cDNA probes should prove useful in studies designed to track cell populations (e.g., tumor metastasis and hemopoietic cells after bone marrow transplantation) in syngeneic male/female pairs. In addition, a cDNA probe that is specific for the rat Sry gene might be valuable in studies of fetal male sexual development or the study of spermiogenesis. PMID- 9203058 TI - Effects of heat stress on mouse testicular cells and sperm chromatin structure. AB - Scrotal regions of mice were exposed to a 38.0, 40.0, or 42.0 degrees C (+/-0.1) H2O bath for 60 minutes to determine the effects of elevated temperatures on testicular cells and sperm chromatin structure. Mice were killed on various days after exposure, and ratios of acridine orange-stained testicular cell populations were determined by flow cytometry. Testicular weights of mice exposed to 42.0 degrees C decreased significantly day 1 (P < 0.01) through 35 (P < 0.001). Also, a significant relative decrease in testicular haploid cells was seen on days 3-35 (P < 0.001) with a corresponding increase in the diploid population (P < 0.001). Testicular analyses of mice exposed to 38.0 degrees C were not significantly different from control values. Testis weights of mice exposed to 40.0 degrees C were not affected, but a relative decrease in percent haploid cells occurred on days 11 and 14 (P < 0.001). The sperm chromatin structure assay (SCSA) was used to measure the susceptibility of cauda epididymal sperm DNA to in situ denaturation at low pH. Caudal epididymides of mice exposed to 42.0 degrees C had no sperm. Caudal epididymal sperm from mice exposed to 40.0 degrees C were most susceptible to acid-induced DNA denaturation on days 3 (P < 0.05), 7, 11, and 14 (all P < 0.001). The 38.0 degrees C exposed mice showed some minor sperm chromatin abnormalities at later time points (days 11-35). When compared to sperm head morphology measurements, SCSA parameters were more sensitive indicators of heat-induced sperm abnormalities. These results show that mouse spermatogenesis is disrupted by scrotal exposure to environmental temperatures several degrees over normal physiological temperature and, of more biological interest, that some thermal ranges above normal allowed production of sperm with compromised nuclear chromatin structure. PMID- 9203059 TI - Seminal plasma lactoferrin concentrations in normal and abnormal semen samples. AB - Although the iron-chelating protein lactoferrin is secreted by the seminal vesicles, the precise role of lactoferrin in semen is unclear. This study aimed to determine whether there is any association between seminal lactoferrin concentrations and normal and abnormal semen samples with and without leucocytospermia. Lactoferrin concentrations were measured by radial immunodiffusion of semen samples from 368 men attending a regional andrology referral center. Routine seminal analysis, including the presence of leucocytospermia, was also performed. Results showed increased seminal lactoferrin in samples showing oligospermia (13.3 mg/100 ml) and oligoasthenospermia (13.4 mg/100 ml) compared to normospermic samples (11.2 mg/100 ml). There were no significant differences in seminal lactoferrin between normospermic samples and azoospermic samples or asthenospermic samples with normal sperm density. Although there was a trend toward increased lactoferrin concentration with leucocytospermia, this was not significant. Possible causes for raised lactoferrin in association with oligospermia are discussed. PMID- 9203060 TI - Copulatory behavior and fertility in transgenic male mice expressing human placental growth hormone gene. AB - Male transgenic (TG) mice overexpressing the human placental growth hormone (GH) variant gene (hGH-V) exhibit reproductive deficits in spite of normal testosterone levels and normal sperm counts. To evaluate the relationship of copulatory behaviors to fertility, we first measured mount, intromission, and ejaculation indices in 2-5-month-old mice (10 TG and 10 normal litter mate controls) during 1 hour tests with ovariectomized, estrogen-, and progesterone primed females. After eight tests, each male was housed with three intact females for 27 consecutive days. Females were checked daily for vaginal plugs and sacrificed 14 days after insemination to determine the numbers of corpora lutea and live and dead fetuses. Relative to their normal siblings, TG mice mounted less often and intromitted sooner after the initial mount, made marginally more intromissions (with and without ejaculation), and were slower to ejaculate. In subsequent fertility tests, TG males inseminated fewer females and sired fewer live fetuses per insemination than non-TG controls. Across TG and normal males, the length of interval between initial mount and initial intromission was inversely correlated with the number of live offspring sired. This suggests that reduced fertility in hGH-V transgenic male mice may be related to altered copulatory behavior, including a rapid progression from first mount to first intromission. PMID- 9203061 TI - Objectively measured boar sperm motility parameters correlate with the outcomes of on-farm inseminations: results of two fertility trials. AB - Two fertility trials were undertaken to evaluate the relationship between boar semen quality and fertility (conception rate and litter size) after on-farm artificial insemination (AI). Trial 1 included 98 ejaculates from 27 boars, and trial 2 included 72 ejaculates from 26 boars. The semen quality was measured by computer-assisted semen analysis (CASA) using the Hobson Sperm Tracker. Boar semen was diluted in a standard extender (Beltsville Thawing Solution, BTS), dispensed into 75 ml allquots each containing 1.5 x 10(9) spermatozoa and dispatched to farms by overnight mail for use by their normal AI procedures. Randomly selected 75 ml aliquots of semen from each boar were also sent to the institute of Zoology for CASA measurement. Prior to CASA analysis, the spermatozoa were recovered from the BTS using Percoll gradients, resuspended in trisbuffered saline media containing 40 mM Ca++, and incubated at 39 degrees C. Parameters of sperm motion were measured after 0, 2, 4, and 6 hours of incubation. Various multiple regression models based on measured motion parameters could account for up to 24% of the variation in litter size. Using logistic regression, highly significant (P < 0.0001) models explaining conception rate in terms of sperm motion were derived for trial 2 only. The change in sperm velocity during the first 2 hours of incubation and the magnitude of the velocity parameters after 2 hours were identified as the most consistent indicators of fertility. Other attributes of sperm quality, i.e., frequency of spontaneous acrosome reactions (AR) and ARs induced by ionophore A23187 or solubilized pig zona pellucida, were also examined. When the "within-trial" median litter size was used as a way of allocating ejaculates to "high" or "low" litter-size groups, higher litter size was associated with lower frequency of both spontaneous and induced AR. These results demonstrate that fertility information can be derived from the CASA analysis of boar semen provided it is combined with a period of incubation in capacitating conditions. PMID- 9203062 TI - Effect of semen dilution on bovine sperm viability as determined by dual-DNA staining and flow cytometry. AB - Living and dead spermatozoa were examined for the effects of sperm concentration level on sperm viability. Semen was collected from two different bulls on each of four collection dates. A ninth bull was collected on all four collection dates as a control for effects of collection date. The ejaculates from these nine bulls were diluted to 30 x 10(6) spermatozoa/0.5 ml and then serially diluted to 20, 10, 5, or 1 x 10(6) spermatozoa/0.5 ml French straw. One-half of the straws for each dilution series was stored 24 hours at 5 degrees C, while the other half was cryopreserved. Spermatozoa were stained with SYBR-14 and propidium iodide (PI) to assess viability. Flow cytometry yielded dot plots showing three distinct sperm populations: dead red-stained spermatozoa (PI), viable green-stained spermatozoa (SYBR-14), and moribund spermatozoa that stained both red and green (doubly stained). Populations were expressed and analyzed in terms of mean percentage of viable spermatozoa and by actual numbers of viable spermatozoa per insemination dose. The mean percentage of living spermatozoa decreased linearly with decreasing sperm concentration; whereas the decrease was parabolic when those same samples were expressed as the mean number of living spermatozoa per insemination dose. The percentage of SYBR-14-stained spermatozoa differed among concentration levels and among bulls (P < 0.01). There were no differences among straws from the same ejaculate. The total volume of ejaculated semen and the concentration of spermatozoa in that ejaculate were both significantly positively correlated with the percentage of SYBR-14-stained spermatozoa in that semen when it was cryopreserved and diluted to < 10 x 10(6) spermatozoa/0.5 ml. In contrast, there were no significant correlations between the initial ejaculate characteristics and the proportion of SYBR-14-stained spermatozoa in the 24-hour stored samples at any concentration. In conclusion, the percentage of viable spermatozoa in an ejaculate significantly decreased with increasing dilution. Further, in cryopreserved samples, the percentage of living spermatozoa < 10 x 10(6) spermatozoa/0.5 ml depended on the original volume and the sperm concentration of that particular ejaculate. PMID- 9203063 TI - Pediatric noninvasive nasal ventilation. AB - Noninvasive nasal ventilation is an effective but underutilized method of chronic respiratory support for patients with respiratory insufficiency due to neuromuscular disease. Noninvasive nasal ventilation corrects nocturnal hypoxia and hypercapnia, resolving symptoms of chronic alveolar hypoventilation. Noninvasive nasal ventilation can allow selected patients with acute respiratory failure to avoid intubation and it can facilitate endotracheal extubation. Practical guidelines and the rationale for pediatric noninvasive nasal ventilation therapy will be discussed in this review. PMID- 9203064 TI - Neurologic onset of Behcet's disease: a diagnostic enigma in childhood. AB - Neuro-Behcet's disease, which is uncommonly reported in childhood, encompasses a wide variety of clinical features since any part of the neuraxis may be involved. It carries a serious prognosis and represents a leading cause of death or severe disability. Neuro-Behcet's disease may occur in 5% to 50% of adults with Behcet's disease and is usually subsequent to other systemic manifestations. In this report, we express the possibility of a primary neurologic presentation of Behcet's disease in childhood with pseudotumor cerebri and meningoencephalitis as exclusive initial features. We focus on diagnostic problems when major features of Behcet's disease are missing at the outset. We emphasize the high sensitivity of magnetic resonance imaging to make the diagnosis of cerebral vasculitis or thrombosis with a good reliability to clinical features. PMID- 9203065 TI - beta-Galactosidase gene mutations in patients with slowly progressive GM1 gangliosidosis. AB - Three unrelated North American cases with slowly progressive forms of GM1 gangliosidosis were found to have two unique point mutations and a 9 bp insertion in the coding region of the gene encoding beta-galactosidase. Case 1 was noted to have a 9 bp insertion ?CAGAATTTT? on one allele between nucleotides 730 and 731 with no other mutations identified in the other allele. In case 2, two point mutations were found: a unique G-->A transition at nucleotide 602 causing an Arg- >His substitution in codon 201 (mutation R201H); and a previously identified G- >T transition at nucleotide 1527 causing a Trp-->Cys substitution in codon 509 (mutation W509C), which has been noted in adult and chronic forms of GM1 gangliosidosis. Case 3 had a unique point mutation (A-->G transition at nucleotide 797) resulting in a Asn-->Ser amino acid substitution in codon 266 (mutation N266S), with no other mutations found in the same or the other allele. Single-strand conformation polymorphism performed on over 100 controls did not demonstrate the presence of the point mutations R201H or N266S. Also, the mutant proteins coded by the two point mutations did not show enzymatic activity in the Cos-1 cell expression system confirming that these mutations are associated with low enzyme activity. PMID- 9203066 TI - Treatment of children with Williams syndrome with methylphenidate. AB - Children with Williams syndrome frequently present with symptoms of attention deficit hyperactivity disorder (ADHD), but there is little information that stimulant medication is useful in this population. A series of double-blind, placebo-controlled case studies was used to evaluate the cognitive and behavioral effects of methylphenidate on four children with Williams syndrome. Teachers and mothers completed behavioral rating scales and cognitive tests of attention, learning and memory, and academic productivity and accuracy in mathematics in each medication condition. Two of the children responded favorably in terms of decreased impulsivity, decreased irritability, and lower activity level as well as improved ability to pay attention. Methylphenidate is a useful adjunct in the treatment of some children with Williams syndrome. PMID- 9203067 TI - Optimizing the indication of vigabatrin in children with refractory epilepsy. AB - This review was conducted to evaluate the long-term prognosis of children responding to vigabatrin by examining the incidence of increased seizure frequency, loss of efficacy, and appearance of new seizures in a cohort of 196 children (mean age, 68.2 months; range, 2 months to 19 years) with drug-resistant epilepsy, who had received vigabatrin as add-on treatment in clinical trials. The results indicate that an increase in seizure frequency was uncommon, occurring in only 10% of children with highly drug-resistant epilepsy and that it usually appears shortly after the initiation of treatment. It was clearly not dose dependent and most often occurred in patients with nonprogressive myoclonic epilepsy. No specific seizure type was specially involved and usually the problem reversed on discontinuing vigabatrin. Loss of efficacy was also uncommon (12% of patients), and again no specific seizure type was found to be associated. Epilepsy syndrome does seem to be a better predictor of loss of efficacy because it occurred most often in symptomatic generalized epilepsies and cryptogenic infantile spasms. A total of 21 patients (11%) developed genuinely new types of seizures. Fifteen of these patients developed new partial seizures that had little impact on the patients' overall clinical improvement. The new partial seizures were better tolerated than the initial seizure type which in most cases had disappeared. Approximately 3% of patients experienced new generalized seizures that aggravated their initial condition. These occurred most often in patients with nonprogressive myoclonic epilepsy; therefore vigabatrin should be used with particular caution in such patients. PMID- 9203068 TI - Respiratory sinus arrhythmia in children with severe cyanotic breath-holding spells. AB - In this study we sought to investigate parasympathetic activity among children with severe cyanotic breath-holding spells by examining respiratory sinus arrhythmia. The study sample was composed of two groups of patients, 16 subjects with cyanotic breath-holding spells (5 male, 11 female; mean age, 37.5 mo) and 17 controls (8 male, 9 female; mean age, 37.7 mo). Each subject's electrocardiogram was recorded in a quiet room and digitized by an 80386 personal computer during five 1-minute periods. R-R intervals within each 1-minute period were converted to heart rate in 120 successive 0.5 second intervals. The resultant heart rate time series was converted to its underlying frequency composition by a fast Fourier transform and averaged across minutes. Respiratory sinus arrhythmia was defined as the variability in the time series over a frequency range (0.096-0.48 Hz) corresponding to a range of respiratory rates from 6 to 30 breaths per minute. Analysis revealed after ANCOVA adjustment for age and gender with subject group and frequency bin as dependent measures, that subjects with cyanotic breath holding spells had similar variability in their heart rates as did controls (group x frequency bin: F = 0.74, P = 0.71). This study supports the hypothesis that autonomic dysregulation in cyanotic breath-holding spells is not due to a primary disturbance in central parasympathetic control over cardiac rate and rhythm. PMID- 9203069 TI - Long-term neurologic and psychomotor sequelae after neonatal tetanus. PMID- 9203070 TI - Acute swelling of the cerebellum on childhood. PMID- 9203071 TI - Idiopathic hypothalamic dysfunction: a paraneoplastic syndrome? PMID- 9203072 TI - Hallervorden-Spatz disease: late infantile type. PMID- 9203073 TI - Relapse and movement disorder after herpes simplex encephalitis. PMID- 9203074 TI - Walther Birkmayer--the man behind the name. PMID- 9203075 TI - Effects of dopamine D-1 and D-2 receptors on intraocular pressure in conscious rabbits. AB - This investigation was designed as a randomized, placebo-controlled, double masked, crossover study in NZW rabbits with normal intraocular pressure (IOP) to investigate dopaminergic effects on IOP. SKF 38393, a selective D1-receptor agonist, increased, and SDZ PSD-958, a selective D1-receptor antagonist, decreased IOP, respectively. The selective D2-receptor agonist quinpirole decreased IOP, whereas the selective D2 receptor antagonist metoclopramide had no significant effect. Combinations of quinpirole with SDZ PSD-958 decreased IOP in an additive manner. SDZ GLC-756, a mixed D1-receptor antagonist/D2-receptor agonist, decreased IOP in a dose-dependent manner with a maximum effect greater than the maximum effects produced either by the D1-receptor antagonist SDZ PSD 958 and the D2-receptor agonist quinpirole. The effect of SDZ GLC-756 could only be partially blocked by the selective D2-receptor antagonist metoclopramide suggesting that both D1-receptor blockade and D2-receptor stimulation participate in its IOP-lowering effect. Tonography suggests that SDZ GLC-756 has no significant effect on outflow facility. Furthermore, the results suggest that both D1 and D2 receptors each play an independent role in the regulation of IOP in rabbits. Thus, simultaneous blockade of D1 receptors and stimulation of D2 receptors may provide a new pharmacological approach for the treatment of ocular hypertension frequently associated with glaucoma. PMID- 9203076 TI - Seizure-related changes in the glutamate R2 and R5 receptor genes expression in the rat hippocampal formation. AB - The expression of mRNA coding for AMPA selective glutamate (Glu) R2 receptor and kainate selective GluR5 receptor was studied in the rat hippocampal formation in two animal models of limbic seizures evoked by systemic administration of pilocarpine (400 mg/kg i.p.) or kainate (15 mg/kg i.p.). As shown by an in situ hybridization study, pilocarpine decreased the GluR2 flip mRNA level in CA1 and CA3 areas of the hippocampus after 3h and kainate after 24h, e.g. at the time preceding neuronal degeneration. No changes in the GluR2 flop or GluR5 mRNA level were found in those regions. In the dentate gyrus, resistant to neurodegeneration, pilocarpine and kainate differentially affected the expression of GluR2 and GluR5 mRNAs. After 72h pilocarpine, but not kainate, increased the GluR2 flop mRNA level and decreased the flip one, which suggests attenuation of the GluR2 sensitivity. On the other hand, kainate, elevated the GluR2 flip and GluR5 mRNA level in the dentate gyrus after 72h. All in all the above data suggest that changes in the GluR2 gene expression may play some role in the neuronal damage to vulnerable areas (CA1, CA3). However, differences in the kainate-and pilocarpine-induced changes in the dentate gyrus at the late time points indicate that alterations in the stoichiometry of GluR2 forms of GluR5 gene expression in this brain region are not a common causal factor responsible for delayed neuronal hyperexcitability. PMID- 9203077 TI - MDMA induced dopamine release in vivo: role of endogenous serotonin. AB - Acting as a substrate at the serotonin (5-HT) transporter, (+)-MDMA (3,4 methylenedioxymethamphetamine), is a potent releaser of 5-HT and causes toxicity to 5-HT neurons after repeated exposure. (+)-MDMA also releases dopamine (DA), although with less potency. Since we have shown previously that the intrastriatal application of 5-HT facilities DA release, it was hypothesized that increased release of striatal 5-HT after MDMA may influence extracellular levels of DA. Using microdialysis in vivo, we found that (+)-MDMA (4.7 mumol/kg, i.v.) administration increased extracellular striatal DA levels to 501% of control (p < 0.01, n = 12). However, in the presence of fluoxetine (14.4 mumol/kg, s.c.), which prevents (+)-MDMA effects on 5-HT release, the (+)-MDMA-induced increase in DA was significantly less (to 375% of control, p < 0.05, vs. no fluoxetine, n = 8). In vitro studies with striatal slices, to test drug selectivity, showed that (+)-MDMA (0.3-3 microM) increased extracellular levels of both DA and 5-HT in a dose-dependent manner. Fluoxetine (3 microM) completely blocked the effects of (+)-MDMA on 5-HT release, but did not alter (+)-MDMA-induced DA release in vitro. The selective DA transport inhibitor GBR-12909 (1 microM), blocked (+)-MDMA's effect on DA release. It is concluded that 5-HT release after (+)-MDMA treatment partially contributes to (+)-MDMA's effect on DA release in vivo. PMID- 9203078 TI - Ex vivo inhibitory effect of the 5-HT uptake blocker citalopram on 5-HT synthesis. AB - Serotonin (5-hydroxytryptamine, 5-HT) synthesis was determined in vivo by measuring the accumulation of 5-hydroxytryptophan (5-HTP) in rat frontal cortex after inhibition of aromatic amino acid decarboxylase by administrative of m hydroxybenzylhydrazine (NSD 1015) (100 mg/kg, i.p.). The selective 5-HT reuptake inhibitor, citalopram, the 5-HT1A agonists, (+/-) 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), ipsapirone, gepirone and the 5-HT1A/B agonist, 7 trifluoromethyl-4(4-methyl-1-piperazinyl-pyrolo[1,2-a]-quinox ali ne (CGS 12066B), the 5-HT1A/B ligands and beta-adrenoceptor antagonists, (+/-) pindolol and (+/-) alprenolol, and the non-selective 5-HT ligands, m chlorophenylpiperazine (mCPP) and metergoline, all inhibited the synthesis of 5 HT. The 5-HT1A/5-HT2 antagonist, spiperone, alone, had no effect on basal 5-HT synthesis, however it attenuated the effect of 8-OH-DPAT by 56% and CGS 12066B by 39% but only barely that of citalopram by 17%. The selective 5-HT1A antagonist, WAY 100635, which did not modify by itself 5-HT synthesis, had no effect on citalopram-induced reduction of 5-HT synthesis. Neither the 5-HT2 agonist, (+/-)1 (2,5-dimethoxy-4-indophenyl)-2-aminopropane (DOI) nor the 5-HT2 antagonist, ritanserin, had any effect on the synthesis of 5-HT. In addition, ritanserin did not modify the inhibitory effect of citalopram. Methiothepin was the only compound to increase 5-HT synthesis. These results suggest that the effect of citalopram on the synthesis of 5-HT is not mediated by 5-HT1A or 5-HT2 receptors and that other receptors may be involved. PMID- 9203079 TI - Prestimulus EEG microstates influence visual event-related potential microstates in field maps with 47 channels. AB - The influence of the immediate prestimulus EEG microstate (sub-second epoch of stable topography/map landscape) on the map landscape of visually evoked 47 channel event-related potential (ERP) microstates was examined using the frequent, non-target stimuli of a cognitive paradigm (12 volunteers). For the two frequent prestimulus microstate classes (oriented left anterior-right posterior and right anterior-left posterior), ERP map series were selectively averaged. The post-stimulus ERP grand average map series was segmented into microstates; 10 were found. The centroid locations of positive and negative map areas extracted as landscape descriptors. Significant differences (MANOVAs and t-tests) between the two prestimulus classes were found in four of the ten ERP microstates. The relative orientation of the two ERP microstate classes was the same as prestimulus in some ERP microstates, but reversed in others. Thus, brain electric microstates at stimulus arrival influence the landscapes of the post-stimulus ERP maps and therefore, information processing; prestimulus microstate effects differed for different post-stimulus ERP microstates. PMID- 9203080 TI - Autoradiographic characterization of [3H]inositol (1,4,5) trisphosphate and [3H]inositol (1,3,4,5) tetrakisphosphate binding sites in human brain. AB - Autoradiographic techniques were used to investigate the characteristics of tritiated inositol(1,4,5)trisphosphate ([3H]IP3) and inositol (1,3,4,5) tetrakisphosphate ([3H]IP4) binding to human brain. In brain sections [3H]IP3 exhibited a two-site binding with KD values of 87 nM and 9.3 microM respectively for the higher and lower affinity sites. [3H]IP4 also bound to two sites with KD values of 43 nM and 1.4 microM, respectively. With the conditions fixed in this study, [3H]IP3 and [3H]IP4 autoradiography in the cortex, caudate, hippocampus and cerebellum were performed. The most prominent [3H]IP3 binding among these regions was found in the cerebellum, particularly in the molecular layer. Within the hippocampus, the subiculum and the CA1 region showed much more prominent binding than the other subfields. [3H]IP4, binding was fairly homogeneous in the regions studied, with the exception of a slightly higher binding in the molecular layer of the cerebellum. PMID- 9203081 TI - Transcranial magnetic stimulation induces alterations in brain monoamines. AB - Transcranial magnetic stimulation has been suggested as a possible therapeutic tool in depression. In behavioral models of depression, magnetic stimulation induced similar effects to those of electroconvulsive shock. This study demonstrates the effect of a single session of rapid TMS on tissue monoamines in rat brain. Alterations in monoamines were selective and specific in relation to brain areas and type of monoamine. The results imply on a biochemical basis to the suggested ECT-like treatment potential of TMS. PMID- 9203082 TI - T-lymphocyte immuno-interferon binding in parkinsonian patients. AB - Human peripheral blood cells, especially lymphocytes and thrombocytes, are extensively studied in neuropsychiatric research both as tools for investigating systemic derangements in neuropsychiatric disorders, and as peripheral models for getting information on central nervous system biochemistry. Specific interferon (IFN)-gamma receptors have been found on both human lymphocytes and neural cells. The aim of the present study has been to evaluate IFN-gamma binding on peripheral blood T lymphocytes from parkinsonian patients, as compared with that on blood T cells from healthy subjects. We have found that T lymphocytes from parkinsonian patients bear a significantly smaller amount of IFN-gamma receptors than those from controls. Such IFN-gamma binding sites are of the same type in patients and healthy subjects (Kd (mean +/- SEM): 1.4 +/- 0.07 vs. 1.2 +/- 0.06, respectively). These findings, which are not specific for Parkinson's disease, are discussed in terms of its immunopathogenesis, since it has been reported that activated T lymphocytes have decreased amounts of IFN-gamma receptors. PMID- 9203083 TI - Attenuation of levodopa-induced toxicity in mesencephalic cultures by pramipexole. AB - The direct-acting dopamine (DA) agonist pramipexole (2 amino-4,5,6,7-tetrahydro-6 propyl-amino-benzthiazole-dihydrochlori de) was evaluated for its ability to attenuate levodopa-induced loss of tyrosine hydroxylase immunoreactive (THir, a marker for dopamine neurons) cells in mesencephalic cultures. Pramipexole reduced levodopa-induced THir cell loss in a dose-dependent and saturable fashion (ED50 = 500 pM), its inactive stereoisomer was significantly less potent in this regard and pergolide and bromocriptine had negligible cytoprotective effects. Culture media from mesencephalic cultures incubated with pramipexole for 6 days increased THir cell counts in freshly harvested recipient cultures. The magnitude of this effect was directly proportional to the amount of pramipexole in the donor cultures and heat-inactivation of the media abolished the growth promoting effect. The results from this exploratory set of experiments suggest that pramipexole may be cytoprotective to dopamine neurons in tissue culture. Pramipexole's affinity for DA receptors, its antioxidant action or its ability to enhance mesencephalic trophic activity could be responsible for this effect. PMID- 9203085 TI - Gait quantitation in Parkinson's disease--locomotor disability and correlation to clinical rating scales. AB - Stride parameters were established in 17 patients with idiopathic Parkinson's disease (PD; mean age 68.8 yrs.; Hoehn-Yahr stages 2 and 3) and in 33 healthy age matched controls. Free-walking speed was lower in PD as were stride length and cadence. Impaired locomotor synergies in PD were reflected by a higher coefficient of variation of stride length; step width and its coefficient of variation (the latter related to postural imbalance in locomotion) were not changed. No stride parameter correlated with any total score of either the Hoehn Yahr Scale, the Unified Parkinson's Disease Rating Scale Motor Examination ("UPDRS-III"), the Columbia Rating Scale (CURS) or the Webster Rating Scale. Stride length correlated with a CURS-Bradykinesia-Score, whereas gait velocity correlated with UPDRS-III-Axial-Motor-Score and with the CURS-Bradykinesia-Score. Hypokinesia of gait in moderately disabled PD patients is best assessed by combined analysis of stride parameters and locomotion-related subscores from conventional rating scales. PMID- 9203084 TI - Effects of tolcapone, a catechol-O-methyltransferase inhibitor, on motor symptoms and pharmacokinetics of levodopa in patients with Parkinson's disease. AB - The effects of tolcapone, a catechol-O-methyltransferase inhibitor, on the bioavailability and efficacy of levodopa were evaluated in 12 patients with Parkinson's disease (PD), 8 of whom showed signs of daily motor fluctuations (wearing-off phenomenon). Motor disabilities were assessed in 12 patients at 7 time points before and after the chronic administration of tolcapone using the Unified Parkinson's Disease Rating Scale (UPDRS). The UPDRS score was improved at all points of determination. Eight patients with wearing-off phenomenon on levodopa showed symptomatic improvement on the combination. The area under the curve (AUC) for levodopa increased by 34% (p = 0.0059) after the administration of tolcapone. The elimination half-life (T1/2) of levodopa was significantly prolonged by 81% (p = 0.0001) after the treatment. The AUC of 3-O-methyldopa, a metabolite of levodopa, was decreased by 79% (p = 0.0001) and the Cmax (maximum concentration) was also decreased by 80%d after the administration (p = 0.0001) of tolcapone. The combination of tolcapone and levodopa was well tolerated. Our findings suggest that tolcapone improves the pharmacokinetics of levodopa in plasma and motor symptoms of fluctuating PD patients. It is suggested that tolcapone may be useful drug adjunct to levodopa in treating patients with PD with wearing-off phenomena. PMID- 9203086 TI - Effect of age on synthesis of the GABAergic steroids 5-alpha-pregnane-3,20-dione and 5-alpha-pregnane-3-alpha-ol-20-one in rat cortex in vitro. AB - Progesterone 5-alpha-reductase activity and 3-alpha-hydroxysteroid dehydrogenase activity were determined in the cortex of male and female rats in vitro. Age effects were investigated. The age of the male rats was 3-23 months, and that of the female rats 4-23 months. On addition, we investigated the enzyme 3 beta hydroxysteroid oxidoreductase, 5-ene-isomerase in rat cortex in order to estimate the local synthesis of progesterone from pregnenolone. We found age-related increases in progesterone 5-alpha-reductase activity in the female rats (r = 0.64, p < 0.01, n = 6) and in the male rats (r = 0.5, p < 0.05, n = 18). 3-alpha HSDH activity remained constant with age in female and male rats. The ratio of 3 alpha-hydroxysteroid dehydrogenase activity to 5-alpha-reductase activity tended to decrease with age (not significantly) in both male rats (r = -0.45, p = 0.06, n = 19) and the female rats (r = -0.36, p = 0.17, n = 16). We could not detect significant metabolism of pregnenolone to progesterone in rat cortex in vitro. The sensitivity of the assays of 3 beta-hydroxysteroid oxidoreductase, 5-ene isomerase was calculated from the mean of the blank values + 3SD; the sensitivity of the assay was calculated as 0.103 fmol/mg protein/min. No significant metabolism of pregnenolone could be detected in cortex pooled from several male rats. The mean metabolism of progesterone was 1,200 times higher than the detection threshold of the assay for 3 beta-hydroxysteroid oxidoreductase, 5-ene isomerase. We conclude that modifications of the inhibitory effects of the GABAergic steroids 5-alpha-pregnane-3,20-dione and 5-alpha-pregnane-3-alpha-ol-20 one via altered progesterone metabolism in rat cortex are possible with aging. A connection with the age-related increase in incidence of epileptic attacks, and with age-related changes in the effects of anticonvulsant and GABAA-active drugs, appears possible. PMID- 9203087 TI - Age-related changes of sodium-dependent D-[3H]aspartate and [3H]FK506 binding in rat brain. AB - We investigated age-related changes in excitatory amino acid transport sites and FK506 binding protein (FKBP) in 3-week-, and 6-, 12-, 18- and 24-month-old Fischer 344 rat brains using receptor autoradiography. Sodium-dependent D [3H]aspartate and [3H]FK506 were used to label excitatory amino acid transport sites and immunophilin (FKBP), respectively. In immature rats (3-week-old), sodium-dependent D-[3H]aspartate binding was lower in the frontal cortex, parietal cortex, striatum, nucleus accumbens, whole hippocampus, thalamus and cerebellum as compared to adult animals (6-month-old), whereas [3H]FK506 binding was significantly lower in only the hippocampus, thalamus and cerebellum. 3[H]FK506 binding exhibited no significant change in the brain regions examined during aging. However, sodium-dependent D-[3H]aspartate binding showed a conspicuous reduction in the substantia nigra in 18-month-old rats. Thereafter, a significant reduction in sodium-dependent D-[3H]aspartate binding was found in the thalamus, substantia nigra and cerebellum in 24-month-old rats. Other regions also showed about 10-25% reduction in sodium-dependent D-[3H]aspartate binding. The results indicate that excitatory amino acid transport sites are more susceptible to aging process than immunophilin. Further, our findings demonstrate the conspicuous differences in the developmental pattern between excitatory amino acid transport sites and immunophilin in immature rat brain. PMID- 9203089 TI - Learning abilities depend on NMDA-receptor density in hippocampus in adult rats. AB - The hippocampal NMDA-receptor is predominantly involved to establish long-term potentiation (LTP) which is assumed to underlie fundamental molecular mechanisms of learning and memory. In the present study, NMDA-receptor density was investigated in parietotemporal cerebral cortex and in hippocampus of commonly bred naive adult male Wistar rats which had performed well or poorly in the passive avoidance paradigm. NMDA-receptor binding was determined in saturation experiments using (3H) MK-801 as a ligand and data for KD and Bmax were calculated from Scatchard plots. In general, higher NMDA receptor density was found in the hippocampus as compared to parietotemporal cerebral cortex. This regional difference became particularly obvious in good performers but was abolished in poor performers. In the hippocampus, a significantly higher NMDA receptor density could be found in rats which had performed well in the passive avoidance task as compared to poor performers. In contrast, no such differences could be found in parietotemporal cerebral cortex. The data may indicate that the reduction in hippocampal NMDA-receptor density is of functional importance, for cognitive abilities in both physiological and pathophysiological conditions. PMID- 9203088 TI - Effects of chronic treatment with a cyclic AMP-selective phosphodiesterase inhibitor, rolipram, on excitatory amino acid neurotransmission systems in young and aged rat brains. AB - Rolipram selectively inhibits cyclic AMP-specific phosphodiesterase, and leads to an increase in cyclic AMP levels in the brain. In this study, we investigated the effects of chronic rolipram treatment on excitatory and inhibitory amino acid neurotransmission systems in young and aged Wistar rat brains. We used in vitro autoradiography with [3H]MK-801, [3H]glycine, D[3H]aspartate, and [3H]muscimol to label N-methyl-D-aspartate (NMDA) receptors, glycine modulatory sites, glutamate transport sites, and gamma-aminobutyric acid-A (GABA) receptors, respectively. Rolipram (0.01 or 0.1 mg/kg, per os) or its vehicle (distilled water) was administered once a day for 4 weeks. The highest binding of [3H]MK-801, [3H]glycine, and D-[3H]aspartate was seen in the hippocampus in vehicle-treated rats. No significant differences in these binding activities were seen between young and aged rat brains. [3H]Muscimol binding was the highest in the cerebellum, and decreased in many brain regions in aged rats. The chronic rolipram treatment resulted in (1) an increase in [3H]MK-801 binding in the dentate gyrus in both young and aged rats, (2) remarkable reductions in D [3H]aspartate binding in many regions of both young and aged rats, and (3) no or minimal changes in [3H]glycine and [3H]muscimol binding. These results suggest that the chronic rolipram treatment modifies the excitatory amino acid neurotransmission system. PMID- 9203091 TI - Inositol has behavioral effects with adaptation after chronic administration. AB - Inositol is a simple dietary polyol that serves as a precursor in important second messenger systems. Inositol in pharmacological doses has been reported recently to be therapeutic in depression, panic disorder and obsessive compulsive disorder. We hereby report effects of inositol on the elevated plus maze model of anxiety. These results should allow development of new inositol analogs that could expand psychoactive drug development possibilities via second messenger manipulation. PMID- 9203090 TI - Effect of sleep deprivation on the growth hormone response to the alpha-3 adrenergic receptor agonist, clonidine, in normal subjects. AB - One night's sleep deprivation (SD) increased the growth hormone (GH) response to clonidine (20 ug/kg i.v.) in 11 normal men ( p < 0.005). This finding may indicate that SD enhances alpha-2 adrenergic receptor function or that the GH response to GH releasing factor in increased by SD. PMID- 9203092 TI - Inositol treatment of autism. AB - Recent studies suggest that serotonin reuptake inhibitors are helpful in at least some symptoms of autism. Inositol is a precursor of the second messenger for some serotonin receptors, and has been reported effective in depression, panic disorder and obsessive-compulsive disorder. However a controlled double-blind crossover trial of inositol 200 mg/kg per day showed no benefit on 9 children with autism. Since biochemical studies could evaluate inositol in children already receiving serotonin reuptake inhibitors. PMID- 9203093 TI - The influence of anatomic and iatrogenic root surface characteristics on bacterial colonization and periodontal destruction: a review. AB - PERIODONTITIS IS A MULTIFACTORIAL infectious disease affecting primarily a subset of subjects and a subset of sites. Recent microbiological data have acknowledged that before disease progression can occur, a susceptible host and site are required, in addition to the presence of pathogenic bacteria. This review discusses factors affecting periodontal disease progression and focuses in particular on the influence of anatomic and iatrogenic root surface characteristics. Retrospective studies clearly suggest a strong association between anatomic aberrations and periodontal attachment loss. Cemental tear seems to have the potential to initiate an aseptic, rapid, site-specific periodontal breakdown in a non-infected environment, illustrating the complexity of the attachment loss process. Recent experimental findings, furthermore, demonstrate a significant influence of root surface instrumentation roughness upon subgingival plaque formation and gingival tissue reactions, as well as a significant and positive relationship between subgingival plaque accumulation and inflammatory cell mobilization. These results indicate that subgingivally located irregularities may form stagnant sites or ecological niches which favor both retention and growth of organisms. Such events in addition to the progressive inflammatory changes may critically influence the subgingival environment by turning a stable site into an unstable or active periodontitis site. Thus, local anatomic and iatrogenic root surface characteristics may have a more profound effect on gingival health than previously assumed, particularly on a site level. PMID- 9203094 TI - Effects of interleukin-1 beta on matrix metalloproteinase-3 levels in human periodontal ligament cells. AB - MATRIX METALLOPROTEINASE-3 (MMP-3), or stromelysin-1, is an enzyme responsible for the degradation of a wide range of extracellular matrix proteins. Increases in MMP-3 activity have been found in several chronic inflammatory diseases, and this increased activity is thought to be mediated by interleukin-1 beta (IL-1 beta). Because IL-1 beta has been strongly associated with inflammatory periodontal disease, the purpose of this in vitro study was to investigate the role of IL-1 beta on the regulation of MMP-3 levels in cells derived from the human periodontal ligament (PDL). Human PDL cell cultures were treated with IL-1 beta at varying concentrations (0.01-1.0 ng/ml) for 24 hour prior to analysis at either transcript or protein levels. Following the isolation of total RNA, the relative levels of MMP-3 mRNA were determined using reverse transcription polymerase chain reaction (RT-PCR) with 32P-end-labeled primers. Immunocytochemical detection of MMP-3 protein was performed using polyclonal antibodies to human MMP-3. The results of RT-PCR analysis demonstrated a concentration-dependent increase in MMP-3 mRNA expression, with IL-1 beta treatments of 0.1 and 1.0 ng/ml significantly (P < 0.01) increased over those cells not treated with IL-1 beta. This increase in mRNA expression was paralleled by significant (P < 0.001) changes at the protein level, with an average of 27.6% of the cells stained positive for MMP-3 following IL-1 beta treatment (1.0 ng/ml), compared with control cells showing no positive staining for MMP-3. In conclusion, the results of this study demonstrate that IL-1 beta upregulates MMP 3 in human PDL cells on both an mRNA and a protein level. These findings suggest possibly important roles for IL-1 beta and MMP-3 in both normal turnover and maintenance of the PDL and in the connective tissue degradation associated with periodontal disease. PMID- 9203095 TI - Increased proliferation, collagen, and fibronectin production by hereditary gingival fibromatosis fibroblasts. AB - HEREDITARY GINGIVAL FIBROMATOSIS (HGF) is a fibrotic enlargement of the gingiva. HGF gingiva contains large amounts of interstitial collagen and other extracellular matrix (ECM) molecules. Increased proliferation and elevated production of the ECM molecules type I collagen and fibronectin (FN) could contribute to the clinical increased bulk of HGF gingiva. Fibroblast strains from HGF gingiva and normal human gingival fibroblast strains (GN) were used in this in vitro study. Fibroblast proliferation was determined by ELISA which measured the incorporation of 5-bromo-2'-deoxyuridine into DNA. The results showed that HGF fibroblast strains proliferated more rapidly than GN fibroblasts (68% to 488% increase, depending on the strains) (P < or = 0.01), the only exception being one HGF strain versus one normal strain. All HGF strains produced greater amounts of FN (measured by ELISA) than all of the normal fibroblast strains (23% to 49% increase, depending on the strain) (P < or = 0.04). Similarly, all HGF strains made significantly greater (P < or = 0.3) amounts of type I collagen (also measured by ELISA) than all of the normal strains (55% to 235% increase, depending on the strain). The results show that, in vitro, HGF fibroblasts display several phenotypic characteristics of activated fibroblasts: increased proliferative rates as well as increased production of FN and type I collagen, consistent with in vitro studies of fibroblasts derived from other types of fibrotic tissue. These results suggest that the increased proliferation of HGF fibroblasts and their increased production of extracellular matrix molecules such as collagen and FN may contribute to the clinical gingival enlargement characteristics of HGF. PMID- 9203096 TI - Reactive change in proliferative activity of the junctional epithelium after topical application of lipopolysaccharide. AB - IT IS WELL ESTABLISHED THAT apical migration of junctional epithelium (JE) along a root surface is an important factor in periodontal pocket formation and deepening. However, the exact mechanism and, more specifically, the role of inflammatory products in influencing the activity of cells within the JE is not known. To address this issue lipopolysaccharide (LPS) was applied topically into rat molar gingival sulcus and then tissues evaluated immunohistochemically for expression of proliferating cell nuclear antigen (PCNA). Tissues were prepared for histological analysis at designated times. Histologically, infiltration of neutrophils with associated edema was noted in JE and gingival connective tissues 6 hours after LPS application and was prominent at 12 hours. These inflammatory changes persisted in the 2- and 3-day specimens, and disappeared at 5 days. In normal gingiva, before the LPS application, the JE showed few PCNA positive cells, while almost all cells in the basal and suprabasal cell layers of the oral gingival epithelium and the oral sulcular epithelium were PCNA positive. No increase in the number of PCNA positive cells in the JE beyond zero time was observed at 6 and 12 hours after LPS application. One day after LPS application, PCNA positive cells appeared in the basal cell layer of the JE, with a continued increase number of PCNA positive cells in the JE continued at 2 and 3 days. By day 5 the number of PCNA positive cells were decreasing with return to a normal range by 7 days. These results showed that 1) under normal physiological conditions, cells within the JE have minimal mitotic activity and 2) the JE cells can enter the proliferating cell cycle when exposed to LPS, and suggest that the enhanced proliferating activity in the JE is an important factor for the deepening of the periodontal pocket, if the connective tissue attachment is broken down. PMID- 9203098 TI - Clinical classification of periodontitis in adolescents and young adults. AB - THE AIM OF THIS STUDY was to determine the degree to which clinical classifications based on cross-sectional assessments endure in the course of development of early-onset periodontitis (EOP), and to introduce new criteria which might improve the clinical classification of these diseases. Subjects with EOP and a matched group without EOP were identified within a national probability sample examined during the 1986/87 survey of US schoolchildren. Of these, 265 subjects (mean age 16 years) were re-examined during the 1992/93 school year. The clinical attachment level of teeth was assessed, and the individuals were classified into localized juvenile periodontitis (LJP), generalized juvenile periodontitis (GJP), incidental attachment loss (IAL), and no-periodontitis groups using three classification methods previously described. A fourth method that considered the extent and severity of attachment loss and the number of missing teeth was introduced to classify the individuals at baseline and at follow-up as having localized, generalized, or incidental EOP, and no periodontitis groups. Furthermore, the individuals were classified using criteria based on the rate and pattern of change in attachment loss during 6 years. The results showed low correlations between the baseline classifications and the classifications at the 6-year follow-up examination, irrespective of the method used. In addition, the cross-sectional classifications were not predictive of the rate of progression of periodontal disease in these subjects. In the generalized disease group, two-thirds of the individuals exhibited moderate/rapid disease progression, while one-third had slow or no progression. In the localized disease group, one-half of the individuals had moderate/rapid disease progression and one half had slow or no progression. In the incidental disease group one-fourth of the individuals had moderate/rapid disease progression and three-fourths had slow or no progression. We propose that the term early-onset periodontitis be used as a generic term to describe periodontal disease before its normal onset. In addition, we suggest that incidental, localized, and generalized EOP are heterogenous groups comprising rapidly and slowly progressing forms within each classification. The findings suggest that a classification system in which subsets of the disease that are defined according to a combination of cross sectional criteria and the disease progression may be useful in studies of EOP. Furthermore, the findings suggest that clinical classifications of EOP be used as generic descriptors until a full understanding of the pathogenesis of this disease is accomplished. PMID- 9203097 TI - Periodontal regeneration in naturally occurring Class II furcation defects in beagle dogs after guided tissue regeneration with bioabsorbable barriers. AB - THE EFFICACY OF A BIOABSORBABLE polylactic acid based barrier was evaluated using naturally occurring buccal Class II furcation defects in beagle dogs. Sixteen furcation sites (8 control and 8 experimental) were treated in 6 adult animals. After full thickness flap reflection, exposed furcations and root surfaces were thoroughly root planed. In experimental sites a customized barrier was formed and fitted to cover the defect. Surgical flaps were replaced slightly coronal to the cemento-enamel junction. Animals were sacrificed at 6 months and specimens processed for histologic evaluation. Histologic and histometric analyses were done using 6 micrograms step serial sections in the buccal-lingual plane, corresponding to the buccal-lingual extent of the furcation. Results were: mean total defect experimental sites 1.92 mm; control sites 1.47 mm. Mean new cementum formation experimental sites 1.36 mm (71% of initial defect); control sites 0.25 mm (17% of initial defect). Mean new bone formation experimental sites 1.42 mm (74% of initial defect); control sites 0.20 mm (14% of initial defect). Mean junctional epithelium formation experimental sites 0.42 mm (22% of initial defect); control sites 1.21 mm (82% of initial defect). Statistical analysis demonstrated significant differences in all healing parameters favoring experimental (barrier-treated) sites. In this model, regeneration (new bone, cementum, and periodontal ligament) of 71% of the original defect in experimental sites and only 14% in control sites demonstrated a response that highly favored use of the barrier. PMID- 9203099 TI - The effect of surface roughness on early in vivo plaque colonization on titanium. AB - THE STUDY ASSESSES IN VIVO the surface roughness necessary to reduce plaque colonization on titanium after 24 hours. Three groups of 16 titanium disks were assigned to 3 different polishing groups (A, B, and C). The roughness was evaluated with a laser profilometer and the morphology with a scanning electron microscope (SEM). Eight volunteers were enrolled and two stents were applied in the mandibular posterior region of each. Each stent supported 3 disks, one per group. The volunteers suspended oral hygiene for 24 hours, after which the stents were removed; one was processed for evaluation of the adherent biomass and the other for SEM study. On each specimen a global area of 100 x 125 microns was examined with SEM. The area was composed of five 20 x 25 microns randomly selected fields. For each field the density of bacteria and the morphotypes were recorded. The data quoted for the global area are cumulative of those observed in the 20 x 25 microns fields. Group A had a significantly smoother surface than groups B and C. The adherent microbial biomass determination and SEM evaluation revealed that group A contained less bacteria than the roughest group. The bacterial population was composed of cocci in group A, and of cocci and short and long rods in groups B and C. We conclude that a titanium surface with Ra < or = 0.088 microns and Rz < or = 1.027 microns strongly inhibits accumulation and maturation of plaque at the 24-hour time period and that such smoothness can be achieved in transgingival and healing implant components. PMID- 9203100 TI - A bone regenerative approach to alveolar ridge maintenance following tooth extraction. Report of 10 cases. AB - TEN PATIENTS WHO REQUIRED two or more anterior teeth extractions were utilized in this study. Extraction procedures were carried out with a full thickness surgical flap approach. After flap reflection, teeth were removed with a minimum of trauma to the surrounding bone. Following extraction silicone-based impression techniques were used to produce a model of the alveolar process and small metal pins were placed in the alveolus to be used as fixed points to make measurements of ridge dimensions. One socket was covered with an expanded polytetrafluoroethylene (ePTFE) barrier membrane (experimental site); the other socket was a conventional control. The soft tissue flaps were then mobilized using periosteal releasing incision and the wound closed with ePTFE mattress sutures. Six months following extraction, patients were treated with flap surgery to expose both extractions sites to remove the ePTFE membranes and to measure ridge dimensions using the pins as fixed points. Clinical and model measurements have shown statistically significant better ridge dimensions at experimental sites than at control (P < or = 0.05). Three patients with exposed membranes had similar dimensional changes as controls. Results from this study suggested that this improved technique offers a predictable alveolar ridge maintenance enhancing the bone quality for dental implant procedures and esthetic restorative dentistry. PMID- 9203101 TI - Guided tissue regeneration therapy of 203 consecutively treated intrabony defects using a bioabsorbable matrix barrier. Clinical and radiographic findings. AB - THE AIM OF THIS RETROSPECTIVE three-center study was to evaluate guided tissue regeneration (GTR) therapy in a clinical periodontal setting. The material consisted of 203 consecutively treated intrabony defects > or = 4 mm in 143 patients using a bioabsorbable matrix barrier. Each center followed the same protocol for presurgical, intrasurgical, and follow up examinations. Initial therapy, surgical, and follow-up treatments followed the routine of each center. Treatment was evaluated after 1 year by clinical assessments for probing depth (PD) reduction and clinical attachment level (CAL) gain and by bone fill from computer digitized radiographs. Initial intrabony defect depth averaged 6.3 +/- 1.0 mm clinically and 5.7 +/- 1.8 mm radiographically. Mean PD was reduced from 9.0 +/- 1.0 mm to 3.3 +/- 1.0 mm. Mean CAL gain amounted to 4.8 +/- 1.5 mm corresponding to 79 +/- 13% of the initial intrabony defect depth; 78% of the defects exhibited CAL gain > or = 4 mm. Bone fill averaged 3.2 +2- 1.8 mm. Together with a crestal resorption of 1.1 +/- 1.4 mm this resulted in a defect resolution of 4.3 +/- 1.9 mm or 72%. Forty-seven percent (47%) of the variability in CAL gain could be explained by defect depth, defect width, early barrier exposure, and presence of plaque in the treated area. CAL gain and bone fill were positively correlated to the intrabony defect depth; i.e., the deeper the defect the more the CAL gain and bone fill. Sites with barrier exposure during the first 2 weeks of healing showed significantly less CAL gain than sites at which exposure occurred at a later stage or not at all. Presence of plaque in the treated area had a significant negative impact on both CAL gain and bone fill. It was concluded that GTR-treatment of intrabony defects > or = 4 mm in a periodontal specialist practice will result in clinical attachment level gain and bone fill comparable to what has been demonstrated in case studies and controlled clinical trials. The predictability to obtain CAL gain > or = 4 mm in defects > or = 4 mm was 78%. PMID- 9203102 TI - Assessment of guided tissue regeneration procedures in intrabony defects with bioabsorbable and non-resorbable barriers. AB - THE PURPOSE OF THIS STUDY was to assess periodontal regenerative techniques in intrabony defects utilizing a bioabsorbable, polylactic acid (PLA) barrier or the non-resorbable, expanded polytetrafluoroethylene (ePTFE) barrier. Thirty patients (26 to 64 years old) each with one radiographically evident intrabony periodontal lesion of probing depth > or = 6 mm participated in a 12-month controlled clinical trial. The subjects were randomly divided into two independent groups. The test group (n = 16) received a PLA barrier. The control group (n = 14) received an ePTFE barrier. Plaque index (PI), gingival index (GI), probing depth (PD), clinical attachment level (CAL), and bone fill were recorded by a single calibrated examiner not involved with the surgical treatment prior to surgery, and at 6, 9, and 12 months postsurgery. The treatment results were statistically analyzed utilizing two sets of data. The "averaged-site" data set consisted of values computed from the averaging of measurements from all sites encompassing the defect. The second data set was comprised of only the deepest measurement of the defect. Statistical tests used to analyze these data sets included the t-test and paired t-test for parametric data and the Wilcoxon rank sum test and the Wilcoxon signed rank test for non-parametric data. Analyses with both the averaged-site data and deepest-site data resulted in significant improvements in PD reductions, CAL, and bone fill, after 12 months of healing with both the PLA and ePTFE barrier devices. Comparisons of healing response between treatments found no significant differences when the averaged-site data were analyzed. When only the deepest site of the defect was considered, the control group resulted in significantly more attachment gain (ePTFE, 3.36 mm; PLA, 1.75 mm; P < 0.02) and shallower probing depths (ePTFE, 3.29 mm; PLA, 4.69 mm; P < 0.01) than the test group. In intrabony defects, the use of PLA or ePTFE barriers in GTR procedures yielded comparable clinical results; however, in this study, data analysis using the deepest site of the defect found, after 12 months of healing, significantly more attachment gain and shallower probing depths with ePTFE. PMID- 9203103 TI - Immediate loading of titanium plasma-sprayed screw-shaped implants in man: a clinical and histological report of two cases. AB - THIS STUDY REPORTS ON THE histological findings of two immediately loaded titanium plasma-sprayed (TPS) implants, retrieved for a fracture of the abutment and for psychological reasons, after 8 and 9 months of loading, respectively. The microscopical analysis showed that mature, compact, cortical bone was present around both implants, with the bone implant contact percentage about 60 to 70%. No fibrous tissue or gaps were present at the interface. No resorption was present in the peri-implant bone. On both implants a few osteoblasts were found positive at the interface for alkaline phosphatase (ALP); while no cells positive for acid phosphatase (ACP) were present. Immediate loading can, perhaps, be used in very selected cases of good bone quality, with implants that have certain macro- (screw shape) and micro-interlocks (titanium plasma-sprayed surface) characteristics. Good results have been reported also for non-TPS surface (e.g., machined surface). More data about different designs (e.g., cylinders) or coatings (e.g., hydroxyapatite) are needed before any firm conclusions about immediate loading can be reached. PMID- 9203104 TI - A clinical evaluation of guided tissue regeneration with a bioabsorbable matrix membrane combined with an allograft bone graft. A series of case reports. AB - THE PURPOSE OF THIS STUDY was to evaluate the clinical effectiveness of a surgical technique in treating periodontal defects. The technique combined tetracycline treatment of a root planed root, grafting of the osseous defect with a demineralized freeze-dried bone allograft combined with tetracycline and the placement of a bioabsorbable matrix membrane, made of polylactic acid softened with citric acid ester. Thirty defects were treated in 27 patients. Statistically significant changes, as a result of the surgical procedure, were observed in marginal recession (mean: 0.5 mm), probing depth reductions (mean: 5.7 mm), and attachment level gain (mean: 5.2 mm). No statistically significant difference existed between the results in the furcation and non-furcation groups. The defects with probing depths > or = 10 mm had a greater mean probing depth reduction (7.4 mm) and mean attachment level improvement (7.2 mm) than the defects with < 10 mm probing depths (probing depth reduction 4.5 mm and attachment level gain 3.9 mm). The proposed surgical procedure seemed to be an effective method to treat periodontal defects. PMID- 9203105 TI - Osseous repair of a lateral periodontal cyst. AB - THE LATERAL PERIODONTAL CYSTS is a slow-growing radiolucent, developmental lesion occurring most frequently in males during the sixth decade. As part of the differential diagnosis, it must be distinguished from the collateral keratocyst and the gingival cyst of adults as well as other entities. Speculation remains as to the lateral periodontal cyst's developmental origin. Whether it is from reduced enamel epithelium, remnants of dental lamina, or cell rests of Malassez remains to be determined. The following longitudinal case report describes the review of literature and clinical and histologic findings as well as unusual treatment of a through-and-through perforating lateral periodontal cyst. Due to the large bony defect left after the cyst's removal, a decalcified freeze-dried bone graft was placed to close the defect. The repair of the lesion was followed for 30 months. PMID- 9203106 TI - Invited essay: the challenge of differentiating normal and disordered personality. AB - By separating personality disorders from other psychiatric conditions and requiring mental health professionals to assess the personalities of all their patients, DSM-III Axis II created an explosion of ideas and research on the nature and structure of personality. Since 1980, theorists and researchers from previously segregated camps have come together to address a number of important taxonomic issues, including the relationship between normal and disordered character. In this article, we place the challenge of differentiating normal and abnormal personality in historical perspective, and outline major theories, models, and methods that inform personologists in their quest. The complexity of personality, and the differences in the way people view the subject matter, ensure that there will be several research lines in the next generation. Progress in the field can be quickened by refinements in theory, the development of more assessment instruments that tap both normal and abnormal traits, and empirical studies that follow well-match groups of normals and patients over significant time periods. PMID- 9203107 TI - Family history assessment of personality disorders: I. Concordance with direct interview and between pairs of informants. AB - The present study examined the concordance of the Family History Interview for Personality Disorders (FHIPD) with diagnoses based on direct interviews and between pairs of informants. Subjects were 224 probands participating in a series of studies of the familial transmission of mood and personality disorders and their first-degree relatives. Proband informants and relatives provided information about themselves on the Structured Clinical Interview for DSM-III-R (SCID), Personality Disorder Examination (PDE), and Eysenck Personality Questionnaire (EPQ). Information from informants about relatives was collected with the FHIPD. All assessments were made blindly and independently. Using Kappa, concordance between proband informants' family histories and relative direct reports on specific personality disorders was low, ranging from -.01 to .28, with a median of .10. Kappa for a diagnosis of any personality disorder was .16. When two independent informant reports were compared, Kappas for specific Axis II disorders ranged from .10 to .72, with a median of .28. Kappa for a diagnosis of any personality disorder was .36. These data suggest that subjects and informants provide different perspectives on Axis II psychopathology, and support the use of both sources of information whenever possible. PMID- 9203108 TI - Family history assessment of personality disorders: II. Association with measures of psychosocial functioning in direct evaluations with relatives. AB - To test the convergent validity of the Family History Interview for Personality Disorders (FHIPD), as well as the general utility of informants' reports of personality disorders, we explored the relationship between proband informant reports of Axis II diagnoses on the FHIPD and relative reports of various indices of psychosocial adjustment. Subjects were the first degree relatives (n = 454) of 224 probands participating in a family study of mood and personality disorders. Relatives provided information on the Structured Clinical Interview for DSM-III-R (SCID), the Personality Disorder Examination (PDE), and other variables reflecting aspects of psychosocial dysfunction that are common in personality disorders. Proband informants were interviewed about their relatives using the FHIPD Proband informant reports of personality disorders on the FHIPD were associated with a variety of forms of psychosocial dysfunction as determined in direct assessments with the relatives, even for those with no diagnosable Axis II psychopathology dysfunction as determined in direct assessments with the relatives, even for those with no diagnosable Axis II psychopathology on direct interview. These results support the convergent validity of the FHIPD, and suggest that informants may provide important information on Axis II psychopathology that is not obtained from direct interviews with the subjects themselves. PMID- 9203109 TI - Self-injurious behavior and mood regulation in borderline patients. AB - This article explores the hypothesis that self-injurious behavior (SIB) of the type associated with borderline personality disorder (BPD) has an important mood regulatory function. Thirty-eight female inpatients with an Axis II diagnosis of BPD and a history of SIB rated a variety of mood and affective states, using visual analog scales recalled over the course of usual SIB experiences. Subjects were additionally divided into two groups according to whether they typically experience pain during SIB (BPD-P group) or did not (BPD-NP group). For both groups, the visual analog scale ratings revealed significant mood elevation and decreased dissociation following self injury, with a peak in dissociative symptoms during self injury. The ratings of dissociative symptoms were found to be higher in the BPD-NP group when compared to the BPD-P group across all stages of SIB. The ratings of sexual arousal did not change over the course of SIB for either group. These findings are discussed in light of current knowledge of the relationship between SIB and mood. PMID- 9203110 TI - Assessing axis II disorders by informant interview. AB - Although much of personality disorder research depends on diagnostic data obtained directly from patients, this approach has rarely been compared to interviews with knowledgeable informants. The purpose of this study was to determine the diagnostic agreement between these two assessment methods, as well as their relative contribution to the formulation of consensus diagnoses. Sixty two psychiatric patients were assessed directly with the Structured Interview for DSM-III Personality Disorders (SIDP), and were asked to nominate an informant- either a family member or friend--to provide information about the patient in an interview with the same instrument. Informant interviews were conducted blind to patient-based information whenever feasible, and diagnostic consensus was achieved by an independent review of all available data by a senior clinician. Diagnostic agreement between patient-based and informant-based personality disorder interview was poor, confirming the findings of two previous studies. Information obtained from patients tended to be given greater weight in formulating consensus diagnoses than information provided by informants. However, about one quarter of diagnostic disagreements were resolved in favor of informant based information. In contrast to a previous study, the inclusion of informant information did not appear to reveal greater psychopathology in patients. We conclude that supplementing direct patient interview with data provided by a knowledgeable informant appears to enhance the resolution of some personality disorder diagnoses. The utility of informant interviews may depend on an analysis of the costs and benefits of this additional degree of descriptive refinement. PMID- 9203111 TI - A psychometric evaluation of the DSM-IV personality disorder criteria. AB - In this study, we examined the psychometric quality of the recently revised DSM IV Personality Disorders (PD). A nationally drawn sample of mental health professionals rated 280 patients known to them, using a symptom checklist containing all the DSM-IV PD criteria. From this symptom level data we determined internal consistency, convergent validity, and discriminant validity for each DSM IV PD criteria. Overall, the findings were encouraging. The internal consistency- a measure of reliability--of the PD criteria sets appears to be substantially improved. The median coefficient alpha for the DSM-IV PDs was .73. Convergent validity was generally adequate for the DSM-IV PD criteria sets. However, discriminant validity, a measure of the specificity of a criterion's relationship to its parent scale, continues to be a problem of the PDs. The findings suggest that the DSM-IV PDs may have better reliability than did their DSM-III-R predecessors. However, the weak discriminant validity found for the DSM-IV PD criteria sets indicates that the Axis II system will continue to show high levels of comorbidity. The usefulness of psychometric procedures in the development and refinement of the DSM PDs is also highlighted. PMID- 9203112 TI - Structured interview versus self-report test vantages for the assessment of personality pathology in cocaine dependence. AB - The study compared structured interview (SCID-II) and self-report test (MCMI-II) vantages for the detection and characterization of personality pathology among 144 urban, poor, cocaine-addicted individuals seeking outpatient treatment. Diagnostic agreement was inadequate for most disorders, and the instruments at best shared only modest common variance. Positive predictive power was poor for all MCMI-II scales, though negative predictive power was good to excellent. This lends support for the use of the MCMI-II as a screening measure to rule out Axis II disorders; however, confirmation of positive diagnoses will require follow-up interview assessment. Future development of self-report personality inventories for substance abusers should focus on controlling for the acute dysphoric effects of drug use and related dysfunction, expanding attention to Cluster B content domains, and incorporating more objective criteria for assessing paranoia and "odd/eccentric" traits. PMID- 9203113 TI - Toward an empirical/theoretical grouping of the DSM-III-R personality disorders. AB - Despite some recent favorable findings, there has not been strong empirical support for the validity of DSM-III-R--and now DSM-IV--personality disorder (PD) clusters. In this study, Axis II symptom ratings on 320 personality disordered patients were used to obtain dimensional scores for the 11 DSM-III-R PDs. The dimensional scores for the DSM PDs were subjected to a principal component analysis with orthoginal rotation. Three factors emerged having eigenvalues > or = than 1. The pattern of factor loadings for the individual PDs were not consistent with the DSM Cluster. Rather, the factor loadings were quite consistent with Millon's theory of personality. These results are discussed in light of the clinical benefits provided by employing empirically determined and theoretically anchored model for organizing the Axis II disorders. PMID- 9203114 TI - Physiological alcohol dependence as a "specifier" of risk for medical problems and relapse liability in DSM-IV. AB - OBJECTIVE: This study tested the ability of DSM-IV physiological alcohol dependence to predict multiple indices of medical problems and relapse behavior. It also tested the ability of three additional variables--DSM-IV nonphysiological dependence, an alternative dichotomous criterion for coding physiological dependence and a dimensional measure of physiological dependence--to predict medical problems and relapse behavior in alcoholism. METHOD: A heterogeneous group of 365 patients was recruited from eight addictions treatment programs in the northeastern United States. A multidimensional assessment battery able to diagnose the presence of physiological dependence according to each of three systems--the criteria of DSM-IV, alternative dichotomous criteria and a dimensional scale-- was administered about 2 weeks after admission, and 241 subjects were reinterviewed 6 months later. The three systems were compared for their ability to predict a variety of external measures of medical complications and relapse liability. RESULTS: Physiological alcohol dependence as diagnosed by DSM-IV bore no relationship to either risk for medical problems or relapse behavior. Further analyses showed that this failure was due to operational problems of physiological dependence in DSM-IV, rather than to a lack of conceptual merit for physiological dependence per se as a course specifier. Use of alternative criteria for coding physiological dependence which are difficult and less internally consistent, and use of a dimensional measure, found improved relationships with the external validators. CONCLUSIONS: Contrary to early reports, physiological dependence can serve as a course specifier for alcohol problems, but must be more sensitively scaled than it was in DSM-IV. Tests of alternative options suggest that a multistage criterion to replace DSM-IV's dichotomous criterion is the best remedy. PMID- 9203115 TI - Pretreatment readiness for change in male alcohol dependent subjects: predictors of one-year follow-up status. AB - OBJECTIVE: The objective of this study was to test a model using pretreatment readiness for change scores (Contemplation, Determination and Action) to predict 1-year alcohol use and recovery activities (AA affiliation and having a sponsor). METHODS: Subjects were 125 middle-aged, mostly white, high school educated, mostly unemployed, male patients who met DSM-III-R criteria for alcohol dependence. The patients participated in a 21-day, Minnesota model, inpatient treatment program. Pretreatment readiness for change was assessed using a modified version of the Stages of Change Readiness and Treatment Eagerness Scale (SOCRATES). The prediction model also included four pretreatment variables: DSM III-R criteria, alcohol consumption, AA affiliation and presence of a sponsor. Logistic regression procedures were used to test the model. RESULTS: High Action scores (and not having a sponsor at pretreatment) were predictive of no reported alcohol use at any time during the 1-year posttreatment period. High Determination scores (and low DSM-III-R criteria scores) were predictive of reported affiliation with AA. Having a sponsor at pretreatment and low Contemplation scores were predictive of reports having a sponsor at follow-up. There were no relationships between the pretreatment readiness for change measures and the actual quantity and frequency of alcohol consumed. There were indications that at follow-up those patients who affiliated with AA or had a sponsor consumed less alcohol than those patients who did not affiliate with AA or have a sponsor. CONCLUSIONS: These results demonstrated a relationship between pretreatment readiness for change and both the decision to drink and to engage in recovery activities; however, it appeared that, once drinking begins, variables other than pretreatment readiness for change influence frequency and quantity of alcohol consumption. PMID- 9203116 TI - The reliability of Form 90: an instrument for assessing alcohol treatment outcome. AB - OBJECTIVE: Project MATCH is a randomized clinical trial consisting of five outpatient and five aftercare units at nine sites. Of importance in this multisite trial examining the efficacy of client-treatment matching was the cross and within-site reliability of the structured interview used to assess alcohol treatment outcomes, the Form 90. Evaluation of the reliability of Form 90 is the subject of this article. METHOD: The reliability of Form 90 was evaluated in two test-retest studies. The cross-site reliability study consisted of 70 paired test retest interviews conducted by different interviewers. Clients for this study were recruited from inpatient, outpatient and college settings. The within-site reliability study had a total of 108 paired test-retest interviews, with 54 of the retests conducted by different interviewers and 54 by the same interviewer. Clients for this study were most often presenting for alcohol treatment at the nine sites and were selected to be representative of the larger Project MATCH sample. RESULTS: Good-to-excellent reliability was found for all key summary measures of alcohol consumption and psychosocial functioning, and most frequently used illicit drugs had moderate reliability. No decay in consistency of self reported drinking was found at more distal points from dates of test-retest interviews. Application of 68% confidence intervals for primary alcohol consumption measures suggests that trained researchers and clinicians can obtain consistent information regarding client drinking. CONCLUSIONS: Form 90 appears to be a reliable instrument for alcohol treatment assessment research when interviewers have received careful training and supervision in its use. PMID- 9203117 TI - Barriers to alcoholism treatment: reasons for not seeking treatment in a general population sample. AB - OBJECTIVE: The present study reports responses to numerous direct questions related to reasons for not seeking alcoholism treatment given the perceived need for treatment among respondents classified with an alcohol use disorder (N = 964, 69.8% male, 93.5% nonblack) in a larger representative sample of the United States population. METHOD: Data were derived from the 1992 National Longitudinal Alcohol Epidemiologic Survey, a national probability sample of 42,862 respondents, aged 18 years and older, from the noninstitutionalized population of the contiguous states. RESULTS: Lack of confidence in the alcoholism treatment system and its effectiveness, stigmatization and denial were identified as significant barriers to alcoholism treatment at the aggregate level. In general, enabling factors such as lack of financial resources or facilities for child care were much less important barriers to care than were individual predisposing factors including attitudes towards alcoholism treatment. CONCLUSIONS: Important sociodemographic differences in identified barriers to care are discussed in terms of their minimization through proposed changes in education, screening, outreach, detection, and referral patterns in alcoholism treatment delivery systems. PMID- 9203118 TI - The contribution of drinking patterns to the relative risk of injury in six communities: a self-report based probability approach. AB - OBJECTIVE: This article reports the results of an analysis of the relationships between demographic and alcohol consumption variables and the likelihood of injury occurring during the 6-month period prior to survey administration. METHODS: The date examined are from a general population survey administered to 22,626 respondents (56.9% female) as part of a community trial project to reduce alcohol-involved injury currently being conducted in six communities in California and South Carolina. Three models that relate consumption patterns and exogenous background variables to injury are evaluated. The first considers only demographic background variables. The second model adds three consumption measures (i.e., frequency, average drinks per occasion and variance) to the first. The third adds a control for community of residence. A fourth model is estimated using frequency and average drinks per occasion, by omitting variance. RESULTS: Findings indicate that likelihood of injury is affected by demographic, alcohol consumption and community of residence variables. Specifically, likelihood of injury is significantly related to being young, white, male and single, and having a high variance drinking pattern. Community level effects were found, with California respondents being more likely to experience injuries, controlling for other model variables. Although measures for education and income were included in our analysis, no effects for these variables could be found. Also, other drinking measures (i.e., drinking frequency and average drinks per occasion) were not found to be significantly related to injury, although estimated coefficients were in the predicted direction. CONCLUSIONS: Complementary to studies that have noted that a large portion of injuries involve persons who have been drinking prior to injury, our findings suggest a link between likelihood of injury and general drinking patterns. PMID- 9203119 TI - A descriptive analysis of the structure and temporal pattern of voluntary ethanol intake within an acquisition paradigm. AB - OBJECTIVE: The present experiment examined the microstructure and temporal pattern of consummatory behavior to provide insight into the behavioral processes that regulate the acquisition of voluntary oral ethanol intake. METHOD: A microcomputer-controlled data acquisition system was used to dynamically monitor food, water and ethanol intake in Long Evans rats across acquisition of ethanol drinking initiated through the presentation of a sequence of increasing concentrations of ethanol solutions in a free choice with water. RESULTS: The results showed a biphasic pattern of ethanol intake as a function of presentation of increasing concentrations of ethanol. Total ethanol intake decreased as the ethanol concentration was increased from 2% to 6%, while, inversely, ethanol intake was significantly increased as the concentration went from 6% to 10%. The initial decrease in ethanol intake, across 2% to 6% ethanol, was a function of decreases in both frequency and size of ethanol bouts. The increase in ethanol intake observed following presentation of higher ethanol presentations was solely a function of increased size of ethanol bouts. The increased size of ethanol bouts was paralleled by an increase in the rate of intake which was not evident across presentation of concentrations below 6%. The pattern of intake across the 23-hour daily sessions exhibited no differences across the dark/light cycle in ethanol or water intake as the concentrations of ethanol were increased. The results indicated, however, that food intake was characterized by increases in consumption during the first hour following the presentation of fluids and the night portion of the dark/light cycle. CONCLUSIONS: The present study revealed, for the first time, the involvement of differential, concentration dependent, behavioral processes in the mediation of the acquisition of voluntary ethanol intake. PMID- 9203120 TI - Renal effects of alcohol withdrawal in five-week alcohol-treated rats. AB - OBJECTIVE: The effects of alcohol withdrawal on renal function following renal ischemia was examined in rats fed a liquid containing ethanol for 5-week alcohol treatment. METHOD: For alcohol-treated rats, animals were fed with an ethanol containing diet for 5 weeks. In withdrawal studies, the alcoholic diet was replaced by a regular diet following 5-week alcohol treatment. Renal ischemia was induced by clamping the renal artery for 20 minutes and renal function was evaluated 24 hours later. RESULTS: Alcohol ingestion for 5 weeks did not alter the renal function in the absence of renal ischemia. Mean (+/- SD) glomerular filtration rate (GFR) and renal plasma flow rate (RPFR) measured 24 hours after ischemia in control rats were 430 +/- 29.6 microliters/min/g/kidney weight (gKW) and 1.4 +/- 0.17ml/min/gKW, whereas in alcohol-treated rats, they were 117.2 +/- 35.2 microliters/min/gKW and 0.31 +/- 0.12ml/min/gKW, which values were significantly lower than controls (p < .05). However, when alcohol was withdrawn for 1 week, the renal function of rats after ischemia was no different from that of control rats (GFR = 413.9 +/- 66.3 microliters/min/gKW and RPFR = 2.14 +/- 0.7 ml/min/gKW). As for renal histopathology, tubular damage was milder 1 week after alcohol withdrawal compared to that observed in rats fed the alcohol-containing diet for 5 weeks. CONCLUSIONS: The findings suggest renal damage induced in rats by exposure to alcohol for 5 weeks was reversed when alcohol was withdrawn for 1 week before renal ischemia. PMID- 9203122 TI - Factors affecting agreement between alcohol abusers' and their collaterals' reports. AB - OBJECTIVE: Because of their low cost and ease of use, collaterals' reports are the most frequent source of independent corroboration with alcohol abusers' self reports of drinking and related events. Although several reviews have shown that we can have confidence in the accuracy of alcohol abusers' reports of their drinking and in the use of collateral reports as an independent validity criterion, neither data source is error free. This study examined factors that influence the level of agreement between collaterals' and alcohol abusers' reports. METHOD: Using data from a study of natural recoveries from alcohol related problems, this study examined how agreement between 120 alcohol abusers' (79.2% male) and their collaterals' reports varied as a function of collateral type and of the collaterals' ratings of their confidence in the accuracy of their reports of the subjects' drinking and related behaviors. Collaterals' awareness of nonalcohol-related levels was also examined. RESULTS: The best agreement occurred for reports from alcohol abusers' spouses who were fairly confident about the information provided. For all variables, some proportion of collaterals respond to demand characteristics of the interview by providing very specific information about subjects' behavior yet admit to being unsure of this information. CONCLUSIONS: Collaterals who are fairly sure of the information they provide are the preferred informants to corroborate alcohol abusers' reports of drinking and related behaviors. In some cases the best collaterals are spouses who are fairly sue of the information they reported. It is also recommended that treatment outcome studies should accept reports only from collaterals who are confident about the information they report. PMID- 9203121 TI - The relationship between Self-Rating of the Effects of alcohol and alcohol challenge results in ninety-eight young men. AB - OBJECTIVE: The level of intensity of response to a drug is likely to influence the future pattern of intake of the substance. This article evaluates a simple Self-Rating of the Effects (SRE) of alcohol form, and reports the relationship between a person's estimate of the amounts of alcohol usually required for four possible effects during three different time frames and his subjective feelings reported during an alcohol challenge. METHOD: SRE forms and results of a challenge with 0.9 ml/kg (0.72 g/kg) of ethanol were available for 18 to 29 year old drinking, but not alcohol dependent, men (N = 98). A subset of 40 subjects completed a second SRE form approximately 1 year later. RESULTS: The correlation between the two SRE administration was .82 (p < .0001), and the results on the SRE were internally consistent, with a higher number of drinks associated with more intense alcohol effects. Focusing on the subjective feelings reported at the 60-minute timepoint during the alcohol challenge, 11 of the 12 alcohol effect categories on the SRE correlated in the predicted direction, including eight that were statistically significant. Evaluating all seven timepoints during the drinking experiment, the average number of drinks on the SRE correlated significantly with the Subjective High Assessment Scale (SHAS) total score at all but the final timepoint. Sons of alcoholics and controls demonstrated similar levels of correlation between SRE and alcohol challenge results. Finally, the SRE correctly identified 79% of the individuals whose levels of response to alcohol fell into the lowest third of intensity during the alcohol challenge, and it correctly classified 60% to 67% of the alcohol challenge subjects who did not fall into that low response category. CONCLUSIONS: The SRE is a simple and reliable measure of a person's estimate of the number of drinks required to achieve a response. The form might be helpful in educating people about the intensity of their response to alcohol and might be useful as a point of discussion in curricula focusing on genetic aspects of alcoholism. When alcohol challenges are not possible in a research protocol, the SRE might help identify a less heterogeneous subgroup of individuals at high risk for alcoholism who have a common mechanism increasing their vulnerability. PMID- 9203123 TI - The role of childhood stressors in the intergenerational transmission of alcohol use disorders. AB - OBJECTIVE: This study examined relations between childhood stressors (e.g., disrupted family rituals, embarrassment, neglect, abuse), family history of paternal alcoholism, and alcohol use disorders in late adolescence and early adulthood. Of particular interest was the extent to which stressor exposure mediated the association between paternal and offspring alcohol use disorders. METHOD: A mixed-gender sample of 457 (238 female) participants, approximately half (N = 234) with a family history of paternal alcoholism, were assessed for alcohol use disorders and childhood stressors via clinical interviews. RESULTS: Family history of paternal alcoholism was associated with every childhood stressor examined, often strongly. In addition, a number of childhood stressors (e.g., verbal, emotional, physical and sexual abuse) were related to an alcohol use disorder in late adolescence/early adulthood. However, only a portion of the effect of family history on a subsequent alcohol use disorder was accounted for by the childhood stressors we examined. CONCLUSIONS: Findings indicate that self reported childhood stressors are strongly related to a family history of alcoholism, but are only moderately and inconsistently related to the development of an alcohol use disorder. Moreover, they appear to, at best, only partially mediate the relation between family history of alcoholism and an alcohol use disorder. PMID- 9203124 TI - Vigilance and iconic memory in children at high risk for alcoholism. AB - OBJECTIVE: Previous studies report reduced visual event-related potential (ERP) amplitudes in young males at high risk for alcoholism. These findings could involve difficulties at several stages of visual processing. This study was aimed at examining vigilance performance and iconic memory functions in children at high risk or low risk for alcoholism. METHOD: Sustained vigilance and retrieval from iconic memory were evaluated in 54 (29 male) white children at high risk and 47 (25 male) white children at low risk for developing alcoholism. Children were also grouped according to gender and age (younger: 8-12 years; older: 13-18 years). RESULTS: No differences is visual sensitivity, response criterion or reaction time were associated with risk status on the degraded visual stimulus version of the Continuous Performance Test. For the Span of Apprehension, no differences were found due to risk status when only 1 or 5 distractors were presented, although with 9 distractors a significant effect of risk status was found when it was tested as an interaction with gender and age (decreased accuracy for older high-risk boys compared to older low-risk boys). CONCLUSIONS: These findings suggest that ERP deviations are not attributable to stages of visual processing deficits, but represent difficulty involving more complex utilization of information. Implications of these results are that the differences between high- and low-risk children that have been reported previously for visual ERP components (e.g., P300) are not attributable to deficits of attentional or iconic memory mechanisms. PMID- 9203125 TI - Correlates of alcohol, tobacco and marijuana use among Scottish postsecondary helping-profession students. AB - OBJECTIVE: There is limited information about the prevalence of recreational drug use over the postsecondary experience in Scotland. The purpose of this study was to investigate the patterns of alcohol, tobacco and marijuana use in postsecondary helping-profession students (medical, nursing, education and psychology) in Scotland in regards to gender, age and course of study. METHOD: The Queensland Alcohol and Drug Study Questionnaire was completed by students enrolled in helping-profession courses from 22 departments at universities and colleges in five Scottish cities. The sample consisted of 717 male and 2,537 female students. RESULTS: A slightly (p < .05) higher percent of women (92.7%) consumed alcohol compared to men (90%), but men consumed significantly (p < .001) more drinks per week (26.7) compared to women (17.3). There was no difference between the two groups when U.K. recommendations of maximum limits for each gender were considered. About 50% of men and women consumed over 21 drinks and 14 drinks per week, respectively. A higher (p < .05) percent of men (42.5%) smoked compared to women (36.9%) and a higher (p < .001) percent of men (40.1%) consumed marijuana compared to women (24.1%). There was no difference in the quantity of tobacco consumed. For both men and women, the prevalence of alcohol and marijuana was highest 2 or 3 years before the maximum use of tobacco (students over 24 years of age). Male and female psychology students consumed the most marijuana. Psychology students, together with nursing students, also consumed the most tobacco. CONCLUSIONS: In view of the increasing prevalence of tobacco over the university experience, especially among nursing and psychology students, and heavier alcohol consumption among younger students, health education programs for Scottish postsecondary helping-profession students should expand from the recently introduced school programs. PMID- 9203126 TI - Altered response in cutaneous sympathetic outflow to mental and thermal stimuli in primary palmoplantar hyperhidrosis. AB - Skin sympathetic nerve activities (SSNAs) were recorded simultaneously from the tibial and peroneal nerves by microneurography at an ambient temperature of 25 degrees C in five subjects with primary palmoplantar hyperhidrosis. The resting of the tibial SSNA innervating the sole (glabrous skin) increased moderately (36.5 +/- 1.5 bursts/min), while mental arithmetic provoked marked responses (1,003.3 +/- 457.4% compared with the resting level) in the hyperhidrosis group compared with the control normohidrosis group (n = 5, 25.3 +/ 4.2 bursts/min and 142.2 +/- 58.4%, respectively). Differentiation of the tibial SSNA into sudomotor (innervating sweat glands) and vasoconstrictor (innervating presphincter of skin vessels) revealed that this SSNA enhancement was attributable to not only sudomotor but also vasoconstrictor components during mental arithmetic. In contrast, the responses in the peroneal SSNA (innervating the dorsum pedis, hairy skin) of the hyperhidrosis group were only slightly changed, exhibiting no significant difference from those in the normohidrosis group. Reflex bursts elicited by sound and electric stimulation were normal in amplitude and latency. When the ambient temperature was elevated to 30 degrees C, the tibial SSNAs became more enhanced than did the peroneal SSNAs. The tibial SSNA was markedly enhanced in the hyperhidrosis group (290.0 +/- 78.5%) compared with the normohidrosis group (78.3 +/- 25.4%). We conclude that the excessive responses in SSNA to the plantar glabrous skin to both mental and thermal stimuli may be responsible for the profuse sweating in subjects with primary palmoplantar hyperhidrosis. PMID- 9203127 TI - Anatomy of medullary and peripheral autonomic neurons innervating the neonatal porcine heart. AB - Gross and microscopic anatomical investigations were carried out in 14 piglets aged from 4 to 66 days. True Blue (7-50 microliters) and Diamidino Yellow (7-50 microliters) were injected individually into 2 different cardiac sites (the right atrial ganglionated plexus, the inferior vena cava, inferior atrial ganglionated plexus, the right atrium or the right ventricle). Gross anatomy: Globular superior cervical and nodose ganglia, elongated stellate ganglia, multiple small middle cervical ganglia and multiple small mediastinal ganglia along the course of cardiopulmonary nerves were identified. Microscopic anatomy: Neurons innervating specific cardiac regions or intrinsic cardiac ganglionated plexuses were distributed relatively evenly among stellate (primarily in their cranial poles) and middle cervical ganglia bilaterally, fewer labeled neurons being located in the superior cervical and mediastinal ganglia bilaterally. Parasympathetic efferent preganglionic neurons associated with either intrinsic cardiac ganglionated plexus studied were identified primarily throughout the ventrolateral region (the external formation) of the nucleus ambiguus bilaterally. Labeled neurons were also identified throughout the right and left nodose ganglia. Individual neurons did not project axons to different cardiac regions, as no double-labeled neurons were identified. No correlation between age and the numbers and locations of labeled neurons was apparent. Thus, porcine sympathetic efferent neurons which innervate individual cardiac regions, including intrinsic cardiac ganglionated plexuses, lie scattered primarily throughout the right and left mediastinal and middle cervical ganglia as well as the cranial poles of stellate ganglia at birth, apparently changing little during the first 2 months of age. Porcine cardiac parasympathetic efferent preganglionic neurons are located primarily in the external formation of the nucleus ambiguus bilaterally at birth. The numbers of afferent cardiac neurons distributed throughout the nodose ganglia bilaterally also change little during that time. It is concluded that most of the autonomic neurons which innervate the heart are in place at birth. PMID- 9203128 TI - Barosensitive cardioinhibitory neurons in the medulla: comparison of FosB/ChAT positive neurons with CT-HRP-labeled neurons. AB - The purpose of this study was to survey the distribution pattern of barosensitive cardioinhibitory preganglionic neurons in the medulla. This was done using Wistar rats anesthetized with fentanyl/midazolam. After stimulating arterial baroreceptors by blood pressure increase due to phenylephrine, c-Fos, FosB, c-Jun and JunD/choline acetyltransferase (ChAT)-positive neurons were surveyed in the medulla. After placing HRP conjugated by cholera toxin (CT-HRP) on the sino atrial node, CT-HRP-labeled neurons were surveyed in the medulla. In the phenylephrine pressor test experiment, we ascertained that in the target neurons fosB was more sensitive to baroreceptor stimulation than any other immediate early genes such as c-fos, c-jun and junD. Using the FosB/ChAT method, we succeeded in determining sites of the barosensitive cardioinhibitory neurons that had never been found with the c-Fos/ChAT method. The distribution pattern of the FosB/ChAT-positive neurons was compared with that of the CT-HRP-labeled neurons. We found that the distribution pattern of the FosB/ChAT-positive neurons in the dorsal motor nucleus of the vagus nerve (DMX) and ambiguus nucleus (AMB) was similar to that of the CT-HRP-labeled neurons at the level between the caudal end of the area postrema (AP) and the caudal end of the nucleus of the trapezoid body. This suggests that the barosensitive FosB/ChAT-positive neurons in the DMX and AMB mediate the cardioinhibitory baroreceptor reflex. PMID- 9203129 TI - Distribution of several gut neuropeptides and their effects on motor activity in muscularis mucosae of guinea-pig proximal colon. AB - The distribution of peptide-containing nerve fibers and the effect of their neuropeptides on motor activity were studied in the muscularis mucosae of the guinea-pig proximal colon. In the immunohistochemical study, it was shown that the tachykinin (TK)-containing nerve fibers densely innervated the muscularis mucosae. In the superfusion study, three kinds of TKs, i.e., neurokinin A (NK-A), neurokinin B (NK-B) or substance P (SP), enhanced the spontaneous activity on the strips of muscularis mucosae with a tetrodotoxin (TTX)-insensitive manner. Their potency was in the rank order of NK-A > SP. This suggests that the muscle has a predominant NK2 receptor. Calcitonin gene-related peptide (CGRP)-immunoreactive fibers were commonly observed in the muscle. CGRP induced a potent inhibition on spontaneous activity and a concentration-dependent inhibition on the NK-A elicited excitation in the presence of TTX, indicating its direct effect on the receptor in the muscle. On the other had, gastrin releasing peptide (GRP), galanin, neuropeptide Y or somatostatin were more or less immunopositive in nerve fibers, but they had no effect on the motility of the muscle except that GRP sometimes showed a faint increase in spontaneous activity. Neither methionine enkephalin nor gastrin-17/cholecystokinin was immunoreactive and had any effect on the muscle. These neuropeptides other than TKs and CGRP do not seem to be neuromediators of motor activity of muscularis mucosae. The results suggest the possibility that TK-, especially NK-A- and CGRP-containing neurons, participate in the regulation of motor activity of the muscularis mucosae in the guinea-pig proximal colon. PMID- 9203130 TI - Neural mechanism of pupillary dilation elicited by electro-acupuncture stimulation in anesthetized rats. AB - The neural mechanisms to reflex dilation elicited by electro-acupuncture stimulation were investigated in anesthetized rats. Two needles, with 160 microns diameter and about 5 mm apart, were inserted into the skin and underlying muscle of a hindpaw. Repetitive 20 Hz, 0.5 ms electrical pulses at various intensities were used for stimulation for 30s. The pupil size was magnified about 44 times via a microscope and was continuously recorded on a videotape. Electro acupuncture stimulation at more than 0.5 up to 6 mA induced stimulus intensity dependent pupil dilation. These responses were abolished by the severance of the sciatic and saphenous nerve of the stimulated hindlimb. Compound action potentials were recorded from the distal cut end of the tibial of a saphenous nerve following electro-acupuncture stimulation of the hindpaw. The mean threshold of the compound action potentials of the myelinated fibers in saphenous nerves was 0.18 mA, while that of unmyelinated fibers was 3.0 mA. The mean threshold of the compound action potentials of the myelinated fibers in the tibial nerve was 0.20 mA of unmyelinated fibers was 3.3 mA. Severance of bilateral trunks did not affect the response, while severance of the third cranial nerves abolished the responses. In conclusion, electro-acupuncture stimulation applied to the hindpaws of the anesthetized rats induced excitation of myelinated or of both myelinated and unmyelinated afferent fibers of the tibial and saphenous nerve, and involved a reflex response of pupil dilation through the third cranial parasympathetic efferent nerve. PMID- 9203131 TI - Temperature dependency of the vagal chronotropic response in the young puppy: an 'environmental-autonomic interaction'. AB - We investigated the effects of mild hypothermia (34.3 +/- 0.2 degrees C [mean +/- SD]), hyperthermia (40.8 +/- 0.2 degrees C) and hypoxia (PaO2 = 43 +/- 4 mmHg) on the response to heart rate to continuous right vagus nerve stimulation (the 'vagal chronotropic response') in young puppies, aged 5-22 days. Puppies were anesthetized with alpha-chloralose, vagotomized and pre-treated with propranolol (1 mg/kg i.v.) and phentolamine (1 mg/kg, 1-2 mg/kg/h i.v.). Hypoxia (n = 9) did not significantly alter the resting sinus cycle length and did not alter the response of sinus cycle to a 30 s train of 8 Hz right vagal stimulation. Mild hypothermia (n = 8) increased the resting sinus cycle length by 16 +/- 4% and greatly augmented the vagal chronotropic response (from 76 +/- 27% change in the sinus cycle length (normothermia) to 155 +/- 38% (hypothermia)). Both the sinus cycle length and the vagal chronotropic response turned towards pre-hypothermia values with rewarming. Mild hypothermia also increased the negative chronotropic response to 20 micrograms/kg/min i.v. of methacholine (12 +/- 2% change in the sinus cycle length (normothermia) versus 24 +/- 14% (hypothermia)), suggesting that a postsynaptic mechanism is involved in the hypothermia-induced augmentation of the cardiac vagal chronotropic response. In contrast to hypothermia, mild hyperthermia (n = 8) decreased the resting sinus cycle length slightly (-5 +/- 5% change) and significantly attenuated the cardiac vagal chronotropic response (from 88 +/- 28% change in sinus cycle length (normothermia) to 50 +/- 26% (hyperthermia)). These changes were also reversible with the re-establishment of normothermia. This demonstrates that clinically relevant, environmentally-induced changes in body temperature can directly and reversibly modify parasympathetic efferent responses. PMID- 9203132 TI - Attenuation of cardiovascular reactions of vocalized and non-vocalized defence areas of periaqueductal grey following lesions in dorsomedial or ventrolateral medulla of cats. AB - In the pretentorial periaqueductal grey (PAG), the region producing pressor responses, vocalization and other somatic and visceral signs (VPR) of the defence reaction and another region producing pressor responses (PR) were localized by electrical stimulation in adult cats, anesthetized with intraperitoneal chloralose (40 mg/kg) and urethane (400 mg/kg). The pressor responses included increases of systemic arterial pressure (SAP) and heart rate, increases of blood flow in the common carotid and femoral arteries and a decrease of blood flow in the superior mesenteric artery. The VPR was found in a relatively restricted region of the dorsolateral PAG, while PR was found scattered within the dorsal and ventral portions of the lateral PAG. The increase of SAP and the changes of blood flow in the sampled arteries were slightly greater during VPR than PR stimulation. Mild vocalization with a slight increase of SAP but marked increases of carotid and femoral blood flow (vasodilation) could be induced by microinjection of N-methyl D-aspartate (0.2 M, 200 nl) into the VPR and the blood flow increase, particularly that of the femoral artery, was greatly attenuated by atropine (1 mg/kg, i.v.). In order to ascertain the contribution of the medullary pressor areas to the VPR- and PR-induced responses, extensive lesions were made in the dorsomedial (DM) or vetrolateral medulla (VLM) by microinjections of kainic acid (KA, 0.024 M) in 27 of the 42 cats. The resting SAP and blood flow of the three arteries were reduced by lesioning of the VLM more than that of the DM. Responses of SAP and blood flow from activation of the PR and VPR, particularly the latter, were affected more after DM compared to VLM lesioning. These data suggest that, while the pretentorial PAG constitutes the 'defence area,' vocalization is confined exclusively to its dorsolateral region and that both the VLM and DM contribute to the cardiovascular components of defence reactions. The DM appears to have a greater contribution compared to the VLM. PMID- 9203133 TI - Localization of NADPH diaphorase in the thoracolumbar and sacrococcygeal spinal cord of the dog. AB - The distribution of NADPH-d activity and NOS-immunoreactivity in the spinal cord of the dog was studied to evaluate the role of nitric oxide in lumbosacral afferent and spinal autonomic pathways. At all levels of the spinal cord examined, NADPH-d staining and NOS-immunoreactivity were present in neurons and fibers in the superficial dorsal horn, dorsal commissure and in neurons around the central canal. Sympathetic preganglionic neurons in the rostral lumbar segments identified by choline acetyl transferase (ChAT) immunoreactivity exhibited prominent NADPH-d and and NOS-immunoreactive staining; whereas the ChAT immunoreactive parasympathetic preganglionic neurons in the sacral segments were not stained. The most prominent NADPH-d activity in the sacral segments occurred in fibers extending form Lissauer's tract through lamina I along the lateral edge of the dorsal horn to the region of the sacral parasympathetic nucleus. These fibers were prominent in the S1-S3 segments but not in adjacent segments (L5-L7 and Cx1 or in thoracolumbar segments. The NADPH-d fibers were not NOS immunoreactive, but did overlap with a prominent fiber bundle containing vasoactive intestinal polypeptide immunoreactivity in the sacral spinal cord. These results indicate that nitric oxide may function as a transmitter in thoracolumbar sympathetic preganglionic neurons, but not in sacral parasympathetic preganglionic neurons. The functional significance of the NADPH-d positive, NOS-negative fiber bundle on the lateral edge of the sacral dorsal horn remains to be determined. However, based on anatomical studies in other species it seems reasonable to speculate that the fiber tract represents, in part, visceral afferent projections to the sacral parasympathetic nucleus. PMID- 9203134 TI - Computer simulation of the enteric neural circuits mediating an ascending reflex: roles of fast and slow excitatory outputs of sensory neurons. AB - Recent electrophysiological studies of the properties of intestinal reflexes and the neurons that mediate them indicate that the intrinsic sensory neurons may transmit to second order neurons via either fast (30-50 ms duration) or slow (10 60 s duration) excitatory synaptic potentials or both. Which of these possible modes of transmission is involved in the initiation of motility reflexes has not been determined and it is not clear and what the consequences of the different forms of synaptic transmission would be for the properties of the reflex pathways. In the present study, this question has been addressed by the use off a suite of computer programs, Plexus, which was written to simulate the activity of the neurons of the enteric nervous system during intestinal reflexes. The programs construct a simulated enteric nerve circuit based on anatomical and physiological data about the number, functions and interconnections of neurons involved in the control of motility. The membrane potentials of neurons are calculated individually from physiological data about the reversal potentials and membrane conductances for Na+, K+ and Cl-. Synaptic potentials are simulated by changes in specific conductances based on physiological data. The results of each simulation are monitored by recording the membrane potentials of up to 16 separate defined neurons and by recording the summed activity of whole classes of neurons as a function of time and location in the stimulated network. The present series of experiments simulated the behaviour of a network consisting of 18,898 sensory neurons and 3708 ascending interneurons after 75% of the sensory neurons lying in the anal 10 mm of a 30 mm long segment of small intestine were stimulated once. The results were compared with electrophysiological data recorded from myenteric neurons during ascending reflexes evoked either by distension or mechanical stimulation of the mucosa. When transmission from sensory neurons to ascending interneurons was via fast excitatory synaptic potentials, the latencies and durations of the simulated responses were too brief to match the electrophysiologically recorded responses. When transmission from sensory neurons was via slow excitatory synaptic potentials, the latencies were very similar to those recorded physiologically, but the durations of the stimulated responses were much longer than seen in physiological experiments. The latencies and durations of simulated and physiologically recorded responses matched only when the firing of ascending interneurons was limited to the beginning of a slow excitatory synaptic (in this study by limiting the duration of the decrease in K+ conductance). The simulation provided several physiologically testable predictions, indicating that Plexus is an important tool for the investigation of the properties and behaviour of the enteric nervous system. PMID- 9203135 TI - The autonomic nervous system: a Sherringtonian revision of its integrated properties in the control of circulation. PMID- 9203136 TI - Effects of cardiac autonomic imbalance on postnatal changes in the electrocardiographic Q-T interval in conscious swine. AB - The effects of partial autonomic denervation of the heart rate and Q--T interval were examined during maturation in swine. Four groups of newborns were prepared: right stellate ganglionectomy (RSG), left stellate ganglionectomy (LSG), right cardiac vagotomy (RCV) and sham-operated. Swine were studied postsurgically for eight weeks. Unexpected deaths of unknown cause occurred in five of the 20 denervated swine but in none of the 10 sham-operated controls. Prolongation of the Q--T interval was greatest in RSG animals and least in RCV as compared to controls. The results indicate that we have successfully developed an animal model of a long Q--T syndrome in swine. PMID- 9203137 TI - Inactivation of the zebrafish homologue of Chx10 by antisense oligonucleotides causes eye malformations similar to the ocular retardation phenotype. AB - We report the cloning of a zebrafish paired-type homeobox gene, Alx, closely related to the murine Chx10 and the gold fish Vsx-I homeodomain proteins. Alx is first expressed at about 12 h post-fertilization (hpf) when optic vesicles appear. Its expression is restricted to the early retinal neuroepithelium, whereas no signal can be detected in the optic placode. Later, Alx expression follows the differentiation of the neural retina. Inhibition experiments with antisense oligonucleotides resulted in specific eye malformations which are reminiscent of the phenotype of ocular retardation (or) mice, caused by a spontaneous Chx10 mutation. The expression of other developmentally relevant genes such as pax(zf-a), pax(zf-b) and krx-20 was not affected in the antisense treated embryos. PMID- 9203138 TI - A sequence conserved in vertebrate Hox gene introns functions as an enhancer regulated by posterior homeotic genes in Drosophila imaginal discs. AB - The intron of the mouse Hoxa-4 gene acts as a strong homeotic response element in Drosophila melanogaster leg imaginal discs. This activity depends on homeodomain binding sites present within a 30 bp conserved element, HB1, in the intron. A similar arrangement of homeodomain binding sites is found in many other potential homeotic target genes. HB1 activity in Drosophila imaginal discs is activated by Antennapedia and more posterior homeotic genes, but is not activated by more anterior genes. Testing a reporter gene construct with mutated binding sites in mouse embryos shows that HB1 is also active in the expression domains of posterior Hox genes in the mouse neural tube. PMID- 9203139 TI - Overexpression of a zebrafish homologue of the Drosophila neurogenic gene Delta perturbs differentiation of primary neurons and somite development. AB - We describe here the isolation and characterization of a zebrafish Delta homologue (delta D). A PCR fragment was used to obtain overlapping cDNA clones encoding a protein of 717 amino acids with all characteristic features of proteins of this family, a signal peptide, a transmembrane domain, and an extracellular region comprising the DSL domain and eight EGF-like repeats. The gene is transcribed in a complex pattern in the developing nervous system as well as in the hypoblast. Overexpression of this gene following mRNA injections leads to a reduction in the number of islet-I positive cells, which are assumed to be primary neurons, and to various defects in the adaxial mesoderm, as well as in the somites and myotomes. This suggests that delta D, and the Notch signalling pathway are involved in the differentiation of primary neurons within the neural plate, as well as in somite development. PMID- 9203140 TI - Developmental expression pattern of Stra6, a retinoic acid-responsive gene encoding a new type of membrane protein. AB - Retinoic acid plays important roles in development, growth and differentiation by regulating the expression of target genes. A new retinoic acid-inducible gene, Stra6, has been identified in P19 embryonal carcinoma cells using a subtractive hybridization cDNA cloning technique. Stra6 codes for a very hydrophobic membrane protein of a new type, which does not display similarities with previously characterized integral membrane proteins. Stra6, which exhibits a specific pattern of expression during development and in the adult, is strongly expressed at the level of blood-organ barriers. Interestingly, in testis Sertoli cells, Stra6 has a spermatogenic cycle-dependent expression which is lost in testes of RAR alpha null mutants where Stra6 is expressed in all tubules. We suggest that the Stra6 protein may be a component of an as yet unidentified transport machinery. PMID- 9203141 TI - A homeodomain point mutation of the Drosophila proboscipedia protein provokes eye loss independently of homeotic function. AB - The Drosophila homeotic gene proboscipedia (pb: HoxA2/B2 homolog) is required for adult mouthparts development. Ectopic PB protein expression from a transgenic Hsp70-pb minigene (HSPB) results in transformation of adult antennae to maxillary palps. In contrast, most tissues appear refractory to PB-induced effects. To study the basis of homeotic tissue specificity we are isolating and studying mutations that modify dominant HSPB-induced phenotypes. One HSPB point mutation (Arg5 of the homeodomain to His) removes homeotic activity in the mouthparts and antennae, but provokes a dose-sensitive eye loss. We show that eye loss can be induced by PB proteins that no longer effectively bind to DNA. The dose-sensitive eye loss thus appears to be mediated by specific, context-dependent protein protein interactions. PMID- 9203142 TI - Xwnt-2b is a novel axis-inducing Xenopus Wnt, which is expressed in embryonic brain. AB - Xwnt-2b is a novel member of the Wnt gene family and is 73-74% similar to human and mouse Wnt-2 proteins. Starting from stage 15, Xwnt-2b transcripts are localized to a non-contiguous stripe in the anterior neural plate of the Xenopus embryo. In the tailbud, Xwnt-2b is expressed along the dorsoanterior side of the prosencephalon-mesencephalon boundary. At the tadpole stages, the brain-specific expression fades, but the total amount of Xwnt-2b mRNA does not decline due to activation of its expression in non-brain areas. Microinjection of Xwnt-2b mRNA into a ventral blastomere of 4-8-cell embryos results in the formation of complete secondary body axes. These results suggest that Xwnt-2b is a member of the axis-inducing Wnts and that it is involved in brain development and in later organogenesis. PMID- 9203143 TI - A role for Xenopus Gli-type zinc finger proteins in the early embryonic patterning of mesoderm and neuroectoderm. AB - Gli-type zinc finger proteins play important regulatory roles in vertebrate and invertebrate embryogenesis. In Xenopus, the Gli-type proteins XGli-3 and XGli-4 are first expressed in earliest stages of mesoderm and neural development. Transient transfection assays reveal that XGli-3 and XGli-4 can function as transcription repressors. Counteracting the Gli-protein repressor activity by ectopic expression of a fusion protein that contains the Gli-zinc finger cluster connected to the E1A activator domain in Xenopus embryos results in specific morphological alterations in the developing somites and in the central nervous system. Altered expression characteristics for a broad set of molecular markers highlighting specific aspects of mesodermal and neural differentiation demonstrate an important role for Gli-type zinc finger proteins in the early mesodermal and neural patterning of Xenopus embryos. PMID- 9203144 TI - An in vitro study on the effect of branch points on the stability of coronary artery flow. AB - Empirical correlations for the onset of turbulence in pulsatile flow through a straight tube and a 45 degrees T-bifurcation are presented. We pumped three different test fluids of kinematic viscosity 0.008-0.035 cm2 s-1 through four straight tubes 0.4-3.0 cm in diameter and three 45 degree T-bifurcations 0.45-2.2 cm in diameter. A Scotch yoke mechanism provided an oscillatory sine wave flow component of known stroke volume and frequency. To determine transition to turbulence, we adjusted the mean flow independently until we detected signal instabilities from hot film or electrochemical wall shear stress probes. The critical peak Reynolds number was found to correlate with two independent dimensionless groups: the Womersley parameter and the Strouhal number. We derived power low functions of these groups to provide an accurate and convenient method of predicting transition in both straight and bifurcating tubes. When compared to pulsatile flow through the straight tube, the presence of flow separation within the 45 degrees T-bifurcation induced flow instabilities at lower values of the peak Reynolds number. The correlation for the 45 degrees T-bifurcation is also a suitable model for predicting transition at coronary branch points, which we previously studied in an in vitro pulse duplicator. Flow instabilities at coronary branch points may play an important role in atherogenesis. PMID- 9203145 TI - A multivariate time-variant AR method for the analysis of heart rate and arterial blood pressure. AB - This paper approaches the problem of short-term mechanisms that regulate heart rate and blood pressure variability signals, by focusing the evident changes of their frequency content during transients (dynamic situations in which the behaviour of these control mechanisms may vary on a beat-to-beat basis). In this study, we suggest an autoregressive time-variant spectral estimation method, which is able to follow such dynamic changes in the signals. This method has also been extended to a multivariate approach in order to take into account more than one process at a time, and to assess the mutual influences between the different controlling systems. The algorithms successfully tested on simulated series have also been used to analyse series recorded during a vaso-vagal syncope episode in a tilt manoeuvre and a physical exercise stress test protocol. The results show how this method is able to follow the changing dynamics of the signals on the basis of a closed-loop model of their interaction on a beat-to-beat basis. After a proper identification procedure of the blocks forming the model, it is possible, therefore, to obtain the classical spectral parameters and the gain of the transfer function between the signals. Such parameters constitute new time series that describe the physiopathology of the cardiovascular control systems, even during non-stationary epochs. PMID- 9203146 TI - A new method for assessment of changes in retinal blood flow. AB - This study validates the use of residence time distribution (RTD) functions in human subjects to assess changes in retinal flow by using the widely recognized model of flow changes due to oxygen breathing. Changes in retinal blood flow may provide important information for clinical decision-making in several populations, including those with diabetic retinopathy, sickle cell disease and retinitis pigmentosa. Normal volunteer subjects were studied before and after oxygen breathing. After i.v. injection, relative fluorescence was obtained using scanning laser ophthalmoscopy/image processing in all vessel branches (average, 17). For each experiment, 64 frames (2/s) were digitized and were normalized using the RTD method. Vessel diameters were measured using densitometry techniques on fundus photos, where the diameter data made it possible to weight each vessel according to relative cross-sectional area to obtain a true mean circulation time (MCT). MCT increased for the group of subjects when breathing oxygen compared to normal air (P = 0.001), representing a decrease in retinal blood flow. Average MCT increased 2.82 +/- 2.51 s for all subjects, with an increase of 2.93 +/- 2.26 s in repeat trials for one subject. The proposed method uses information from all retinal vessels and allows the assessment of overall, as well as selected, regional retinal flow. It is more comprehensive than previous methods analysing single vessel flow. This method will be potentially useful for assessing hemodynamic changes in the retina associated with a wide range of eye disease. PMID- 9203148 TI - Spatial resolution in plantar pressure measurement. AB - Peaks of pressure under the foot are of significance to the understanding and limitation of damage to plantar tissues. At present, many of the pressure measuring systems use a matrix of discrete transducer cells which each register the average pressure over their surface, and whose dimensions are such that they limit the accurate representation of the true peak pressure. The spatial filtering effect is investigated in this paper by analytical deduction from a pedobarographic record of high spatial resolution. For a barefoot diabetic patient who presents sharp peaks of pressure in the metatarsal region, the effect of national cell dimensions is investigated through analysis of a typical record and shown to constitute a potentially significant error for cells of the dimensions of those in common use. The average pressure read from a transducer of area 100 mm2 may be of the order of 60-70% of the true peak in barefoot standing, although this depends on the sharpness of the peaks. Errors are decreased when the more even in-shoe pressure distribution is considered. PMID- 9203147 TI - Time-dependent stress and displacement of the eye wall tissue of the human eye. AB - Myopia or short sightedness, is the most important predisposing factor to retinal detachment. The relative risk of detachment rises with increasing myopia. The model characterizes that because the severity of myopia increases with the axial length (antero-posterior diameter) of the eyeball, the relative risk of retinal detachment rises with increasing eye size. We present a mathematical model of the time-dependent shear stress force that occurs in the thin eye wall shell supporting the vitreous humour inside the eye globe during the acceleration and deceleration phases of saccadic eye movement. Results show that the shear force increases as the thickness of the eye wall decreases. It is common for myopes to have thinner eye wall tissue than emmetropes. In addition, if account is taken of the increased force required to provide normal saccadic movement of myopic (larger) eyes, then the shear force is up to seven times greater than that experienced for emmetropes. PMID- 9203149 TI - Biomechanics of load-bearing of the intervertebral disc: an experimental and finite element model. AB - This paper presents an experimental and finite element study of the biomechanical response of the intervertebral disc to static-axial loading in which classical consolidation theory was used to analyse its time-dependent response. A newly developed experimental technique was employed to load the disc in compression and measure simultaneously the matrix internal pressure and creep strain for the full consolidation process. It is shown that, upon loading, the disc develops a maximum hydrostatic excess pore pressure which gradually decays as water is exuded from the matrix. During this decay process, the applied load is progressively transferred to the solid components of the matrix until the load is borne in full by the solid at the end of consolidation. Material properties for the tissue were then obtained from the experimental stress-strain data and used in the finite element study in the development of a finite element solution based on Biot's theory of coupled solid-fluid interaction. An axisymmetric formulation was employed and the disc modelled as an anisotropic, non-linear poroelastic solid. A sensitivity analysis of the material properties for the structural components of the disc was carried out and the biomechanical response to compressive loading evaluated and compared to experimental data. The results show that the matrix permeability plays a significant role in determining the transient response of the tissue. Annular disruptions of the disc were shown to result in an increase in the nuclear principal stresses suggesting that disrupted regions of the annulus fibrosus play a reduced role in load bearing. PMID- 9203150 TI - Liquid ventilation: a mathematical model of gas diffusion in the premature lung. AB - The liquid ventilation (LV) technique was previously demonstrated to be a valuable alternative to ordinary gas ventilation, particularly for newborn patients with severely distressed lungs. This work describes a mathematical model of gas transfer phenomena occurring within the lungs of a preterm newborn baby ventilated with liquid perfluorocarbon (PFC) RM-101. The model was conceived in order to perform computer simulations of LV treatments. Its input parameters are tidal volume, respiratory frequency, oxygen and carbon dioxide tension in inlet PFC; its output data are the partial pressures of respiratory gases in the alveolar environment. Such values may be evaluated at any instant from the beginning of the treatment, in order to judge whether the therapy is able to meet the necessary conditions to arterialize properly the patient's venous blood. The model also enables optimisation procedures to be defined and performed. Quantitative results and graphs are supplied, with reference to the simulation of LV applied to a preterm newborn of 28 gestational weeks. The main results point out that a relatively short duration of initial transients is attainable (200 to 240 s) and that blood arterialization is possible even with low oxygen tension in inlet PFC (29.7 kPa (223 mmHg)). PMID- 9203151 TI - A dual coordinate system finite difference method for forward and inverse solutions of the volume conductor problem in neurology. AB - Finite difference methods for the volume conductor problem have used a single coordinate system for the mesh and made approximations of Laplace's equation. This method is simple but has two major problems. Firstly, to deal with boundary conditions properly, the normal potential gradient at the boundary must be known. However it is complicated to compute at a curved surface point. Secondly, for an inverse solution the equation on a curved boundary is difficult to reverse since more than one inner mesh node appears in the approximation equation for each surface point. The new method developed in this paper is a dual coordinate system. One system serves as a frame mesh, the other is a sub-coordinate system in which surface points become mesh points (regular nodes). The equation at each surface point is then directly reversible since only one inner point appears in the equation. The forward solution is applied to both centric and eccentric bone models and uses the conventional successive over-relaxation (SOR) method. Noise is added to this solution for input to the inverse procedure which is a direct step-in non-iterative method. Low pass filtering was effective in reducing the effects of noise. In the examples given, only one coordinate subsystem is used but, for complex shape boundaries, multiple subsystems would be necessary. PMID- 9203152 TI - A scoliosis correction device based on memory metal. PMID- 9203153 TI - Comparison of predicted and measured contact pressures in normal and dysplastic hips. AB - Hip dysplasia, a congenital and developmental deformity characterized by malorientation and a reduction of contact area between the femur and acetabulum, is the most common cause of osteoarthritis of the hip. According to current estimates, dysplasia accounts for nearly 76% of all cases of osteoarthritis, and many who are affected require a total hip replacement before the age of 50. It is theorized that in the poorly oriented and deformed pelvis, a reduction in contact area leads to an increase in contact pressure during normal activities. Currently, clinicians attempt to reposition the joint, assuming that improving the position of the existing contact surface will lead to decreased pressures. It is also assumed that improving certain geometric parameters correlates indirectly with decreased contact pressures. Neither these simple estimates nor other non invasive models have ever been shown to be related to contact pressure. The purpose of this study was to evaluate a computerized method of predicting hip joint contact pressures, which applies known hip joint reaction forces to the three-dimensional surface of the hip joint. To this end, cadaveric and plastic pelvic models were developed to test whether the computer model could predict the magnitude and location of maximum pressure. Mechanical testing revealed that the computer model could be used to predict pressure in cadaveric pelves at prescribed locations (r2 = 0.64). The computerized model could also be used to predict the magnitude and location of maximum pressure in a series of plastic models where the load vector and the degree of dysplasia were parametrically varied (r2 = 0.7). These findings suggest that the computer model may be useful in identifying patients who will fail osteotomy or whether they can be used to select the best osteotomy for each patient. PMID- 9203154 TI - Design and evaluation of a device for measuring three-dimensional micromotions of press-fit femoral stem prostheses. AB - Implant micromotion is considered to be a major factor in the loosening of cementless total hip replacements. Translational micromotion at the bone-implant interface generally occurs in all three spatial directions. Under physiological loading, the interfacial micromotion consists of a cyclic amplitude and changes in the mean, which, in the cranio-caudal direction, represents subsidence of the prosthesis. Existing measurement strategies, which are based on dial gauges, extensometers, LVDTs, hall-effect transducers or strain gauge techniques provide information about only one component of the general three-dimensional micromovement. Moreover, in the majority of the studies, the data are difficult to interpret due to the measured motions being composed of interfacial micromotion and femoral strains. A new transducer was designed that allows the accurate measurement of all three isolated components of micromotion. An optoelectronic approach, based on silicon position-sensitive detectors (PSD) in combination with high precision mechanical parts, was chosen. To exclude thermodrifts during long-term testing, a thermistor was integrated in the sensor. Validation experiments on a precision positioning table indicated the high precision and resolution of the developed sensors. Furthermore, in-vitro tests on a standard press-fit prosthesis demonstrated the easy handling and reliability of the system. PMID- 9203155 TI - Regulation of gene expression in the nervous system by reactive oxygen and nitrogen species. AB - Reactive oxygen and nitrogen species function as direct and indirect modulators of gene expression through their interactions with transcription factors and also key enzymes in receptor-activated signalling pathways. This regulatory role may become displaced under certain circumstances such as aging, autoimmune responses and viral infection, leading to the pathological outcome associated with inflammatory and degenerative diseases in the CNS. PMID- 9203156 TI - Effect of trophic factors on delayed neuronal death induced by in vitro ischemia in cultivated hippocampal and cortical neurons. AB - The effect of trophic factors on neuronal survival after 30 min oxygen and glucose deprivation (in vitro ischemia) was studied in primary hippocampal and cortical neuronal cultures of rat. In vitro ischemia was produced at 37 degrees C by placing cultures in glucose-free medium, the oxygen content of which was removed by gassing with pure argon. After in vitro ischemia neurons were allowed to recover either in serum-free minimal essential medium (MEM) or in MEM containing 5% native horse serum, 100 ng/ml basic fibroblast growth factor (bFGF) or 10 ng/ml transforming growth factor-beta 1 (TGF-beta 1), respectively. Cultures that recovered in serum-free medium suffered a progressive type of neuronal injury: survival of either cortical or hippocampal neurons declined from about 60% after 1 h to 50% after 3 h, 40% after 6 h and less than 20% after 24 h. Addition of serum proteins to the incubation medium did not influence early survival (up to 3-6 h) but significantly improved survival after 24 h (more than 40% in both hippocampal and cortical cultures). Addition of TGF-beta 1 and bFGF had only minor effects. These data show that serum reduces delayed ischemic cell death by a mechanism which is different from that of TGF-beta 1 or bFGF protection. PMID- 9203157 TI - Effects of simulated upper gastrointestinal hemorrhage on ammonia and related amino acids in blood and brain of chronic portacaval-shunted rats. AB - Gastrointestinal (GI) hemorrhage during compromised liver function is known to precipitate portal-systemic encephalopathy (PSE). Hypothetically, the induced hyperammonemia depletes cerebral glutamate pools. To investigate this hypothesis, rats were studied 14 days after portacaval shunt (PCS) or sham surgery (SHAM). Rats received 3 mL bovine erythrocytes or saline at t = 0, 1, 2, and 3h via a previously placed gastrostomy catheter. At t = 0, 2, 4, 6 and 8h arterial blood and at t = 8h cerebral cortex were sampled for determination of ammonia and amino acids. Control rats (NORM) were sampled without previous surgery. Repeated intragastric blood administration increased the already elevated arterial ammonia levels in PCS rats further. This resulted in higher cerebral cortex ammonia and glutamine levels after blood administration. Despite the accumulation of ammonia and glutamine, cerebral cortex glutamate concentrations remained unaltered. Yet, PCS rats became more encephalopathic after blood gavages, suggesting that there is not a clear-cut relation between cerebral cortex glutamate concentrations and degree of PSE. Interestingly, cerebral cortex concentrations of GABA, tyrosine and phenylalanine were markedly increased. Whether these observations are pathogenetically related to PSE remains to be established. The present model of simulated GI hemorrhage in PCS rats seems to be a suitable, clinically valid model for future research regarding hepatic encephalopathy. PMID- 9203158 TI - Increased cerebral free radical production during thiamine deficiency. AB - Concentration of reactive oxygen species (ROS) and the antioxidant glutathione (GSH) was measured in thalamus and cortex after 13 and 14 days of pyrithiamine induced thiamine deficiency (PTD) in the rat. The concentration of ROS was significantly elevated in thalamus and cortex on day 14 when righting reflexes were absent and spontaneous seizures occurred. No significant changes in GSH concentration were observed in thalamus or cortex on either day of treatment. These findings suggest that increased formation of free radicals occurs during the more acute symptomatic stage of thiamine deficiency and may contribute to the structural damage described in this model of Wernicke's encephalopathy. PMID- 9203159 TI - In vivo study of the kinetics of thiamine and its phosphoesters in the deafferented rat cerebellum. AB - The effects of chemical (CD) and surgical (SD) deafferentation of the cerebellum on different steps of the metabolism of thiamine (Th), thiamine monophosphate (ThMP) and thiamine pyrophosphate (ThPP) were evaluated in vivo in rats. CD was carried out by i.p. injection of 3-acetylpyridine, followed by harmaline and niacinamide. SD was carried out by complete dissection of the peduncles of the left cerebellar hemisphere. Under steady state condition the radioactivity of Th and its phosphoesters was determined in plasma and whole cerebellum after an i.p. injection of thiazole-[2(14)C]-thiamine (30 micrograms:1.25 micro Ci). Analytical data were processed by using an improved mathematical compartmental model, which allowed the calculation of fractional rate constants (FRC), turnover rates (TR) and turnover times (TT). Both CD and SD caused a significant reduction of TR values for Th phosphorylation to ThPP, dephosphorylation of ThPP to ThMP and Th, and ThMP, but not Th, release. TT for all Th compounds were increased compared to controls, indicating a general slowing of thiamine metabolism in the deafferented cerebellum. These results indicate an imbalance in the thiamine metabolism resulting from the impaired activity of cerebellar neurons. The possible implications of the changes in rate of Th compound turnover with respect to biochemical changes in cerebellar ataxia are discussed. PMID- 9203160 TI - Measurement of the expiratory ammonia concentration and its clinical significance. AB - Although gaseous ammonia (NH3) can freely enter cells through the plasma membrane where NH3 is cyto(neuro)toxic, NH3 and ionic ammonia (NH4+) contents have not been studied in biological materials. We developed a new method for measurement of expiratory NH3 concentration, which may reflect blood NH3 concentrations. The method is a sensor tube type-gas assay system. Expiratory NH3 concentration in patients with chronic liver diseases increased when their blood ammonia (NH4(+)+NH3) concentrations increased above 90 micrograms/dl (normal range; 12-66 micrograms/dl). However, cirrhotic patients, who had relatively higher expiratory NH3 concentration compared to blood NH3 concentrations (calculated from Henderson Hasselbalch formula), were found to have subclinical encephalopathy. Measurement of expiratory NH3 concentration may be of clinical significance for the diagnosis of encephalopathy associated with hyperammonemia. PMID- 9203161 TI - The effects of unsaturated fatty acids, oxidizing agents and Michael reaction acceptors on the induction of N-ethylmaleimide reductase in Escherichia coli: possible application for drug design of chemoprotectors. AB - Menadione and dimethyl maleate, Michael reaction acceptors, induced N ethylmaleimide (NEM) reductase activity in Escherichia coli strain DH5a. Linoleic acid also induced NEM reductase activity, but oleic acid, which is less susceptible to lipid peroxidation than linoleic acid, did not induce NEM reductase activity. In addition, NEM reductase activity was induced by menadione and linoleic acid also in strain DH5, Y1088 and Y1090. Linoleic acid is not a Michael reaction acceptor, but is known to produce Michael reaction acceptors such as alkenals and 4-hydroxyalkenals as a result of free-radical-initiated lipid peroxidation. Thus, our findings suggested that lipid peroxidation was involved in the induction of NEM reductase by linoleic acid. The electrophilic property of Michael reaction acceptors provides the signal for induction of phase II enzymes such as glutathione S-transferase and quinone reductase in mammals. The inducer potency of phase II enzymes has been used to design chemoprotective drugs. Therefore, the inducible nature of this enzyme will serve not only for the elucidation of its physiological function, but also for the evaluation of chemoprotective drugs. PMID- 9203162 TI - Oxidized low density lipoprotein acts on endothelial cells in culture to enhance endothelin secretion and monocyte migration. AB - Monocyte deposition on the endothelium is the initial step in atherogenesis. Oxidized low density lipoprotein (Ox-LDL) is involved in the development of the fatty streak which progresses to the atherosclerotic lesion. Our interest focussed on the question, does the endothelium react to Ox-LDL to produce humoral substances that might influence the migration of human blood monocytes? Chemotaxis of monocytes was assessed by the modified membrane-filter technique based on the Boyden chamber principle. Exposure of porcine aorta endothelial cells (ECs) to Ox-LDL (100 micrograms/ml) increased the directional migration of monocytes by 25% (p < 0.01) over that of ECs in the absence of Ox-LDL. Radioimmunoassay of the EC culture media revealed the presence of immunoreactive endothelin-1 (ir-ET-1). The endothelin converting enzyme inhibitor, phosphoramidone (10 microM), when incubated together with ECs and Ox-LDL, suppressed the synthesis of ir-ET-1 by 53% (p < 0.05) and the migration decreased by 12% (p < 0.05). Preincubation of monocytes with the ETA receptor-selective antagonist, BQ-123 (1 microM), followed by exposure to ECs plus Ox-LDL, lead to a decrease in their migration by 12% (p < 0.05) compared to monocytes not treated with BQ-123. These results show that Ox-LDL acts on ECs to enhance the synthesis of ir-ET-1 which in turn increases the directional migration of monocytes. Phosphoramidone decreased the synthesis of ir-ET-1 but migration was affected only modestly; monocyte ETA receptor blockade by BQ-123 also suppressed migration toward EC chemoattractants to a small extent. Both results suggest that in addition to ir-ET-1 other chemotactic factors are being released by the ECs; Ox LDL appears to enhance their release or synthesis. PMID- 9203163 TI - Effects of [arginine8, glycine-OH9]-vasopressin on pentylenetetrazol seizures in mice. Interaction with adrenaline. AB - The effects of [arginine8, glycine-OH9]-vasopressin (AGV), alone and in combination with adrenaline, on pentylenetetrazol (PTZ) seizure threshold by timed intravenous infusion in tall vein and intensity by subcutaneous (s.c.) PTZ test (85 mg/kg) were studied in male albino mice. AGV was administered intracerebroventricularly (i.c.v.) at doses of 0.001, 0.01, 0.1, 1.0 and 5.0 ng/mouse 5, 15 and 30 min prior to PTZ AGV induced decrease of seizure threshold at middle doses (0.01 and 0.1) 5 and 15 min prior to PTZ (75 and 67% respectively vs. controls). Adrenaline (1 mg/kg, i.p.) potentiated the effect of AGV on seizure threshold. AGV also induced increase of seizure intensity at doses of 0.01 and 1.0 ng and decrease of latency of the first tonic seizure. Adrenaline (1.0 mg/kg, i.p.) enhanced the effects of AGV suggesting interactions of vasopressin with adrenergic neurotransmission in the CNS. PMID- 9203164 TI - A study on the involvement of GABAB receptor ligands in stress-induced antinociception in male mice. AB - Swimming at 21 degrees C for 3 min induced antinociceptive effects in male mice with all animals showing a significant increase in response time to the hot plate test measured 10 min after swimming. This antinociceptive activity was still evident at 20 min after swimming. Prior administration of the GABAB receptor agonist, baclofen, potentiated the swimming-induced antinociception. For the group receiving the higher dose of baclofen (2.0 mg/kg, s.c.) the potentiation was still evident 40 min postswimming. However, prior administration of CGP 35348, a GABAB receptor antagonist, had no effect on the antinociceptive activity observed after swimming. Restraint for 1 h also induced significant antinociceptive activity in male mice. This restraint-induced antinociceptive activity was enhanced by prior administration of baclofen and was completely abolished by the administration of CGP-35348 before restraint. The present findings suggest that GABAB receptors may play a role in stress-induced antinociception and different GABAB receptor subtypes may be involved depending on the nature of the stress. PMID- 9203165 TI - Effects of cigarette smoke inhalation on plasma diltiazem levels in rats. AB - A rapid and sensitive method for the assay of plasma diltiazem was developed using a solid-phase extraction technique followed by high-performance liquid chromatography. The effects of cigarette smoke on plasma levels of orally administered diltiazem was investigated in rats. The animals were exposed to cigarette smoke for 10 min using a Hamburg II smoking machine, immediately after oral administration of diltiazem (10 mg/kg). In the nonsmoking nonrestrained rats, plasma diltiazem levels increased rapidly and reached the maximum (7.1 micrograms/kg) 2 h after administration and decreased gradually thereafter. In the nonsmoking restrained rats, plasma diltiazem levels increased rapidly, but showed almost constant levels between 1 h and 8 h after administration. The maximum level (5.4 micrograms/kg) was shown after 2 h. On the other hand, plasma diltiazem levels in the rats exposed to cigarette smoke reached the maximum (4.3 micrograms/kg) after 4 h. These results suggest that absorption of orally administered diltiazem is inhibited and delayed by cigarette smoke. PMID- 9203166 TI - Theophylline-induced changes in mammalian adenosine deaminase activity and corticosterone status: possible relation to immune response. AB - Theophylline at low doses (10 mg/kg/day p.o.) under long-term conditions (for 16 consecutive days) increased the adenosine deaminase (ADA) activity in spleen and thymus of adult male albino rats without changing its hepatic ADA activity. Treatment with high doses (20 mg/kg/day p.o.) under similar conditions, on the other hand, decreased the splenic and hepatic ADA activity and increased the thymic ADA activity. This induction of thymic ADA activity, however, was significantly less than that observed with low doses of theophylline. The plasma corticosterone level was increased without changing its adrenal level under similar conditions. This study, thus, indicates that long-term theophylline treatment may potentiate or suppress the immune response, depending on the dose, through the tissue (liver/spleen/thymus)-specific modulation of ADA activity and plasma corticosterone status. PMID- 9203167 TI - Plasma protein binding of levamisole in several species. AB - The binding of levamisole to total plasma proteins of 6 animal species was determined in vitro by equilibrium dialysis. The percentage of bound drug protein was independent of levamisole concentration within the range studied, 5-50 micrograms/ml (ANOVA). Levamisole was bound to a low extent to plasma proteins of each animal species (19.40-25.91%). There were significant differences in the extent of levamisole binding among species (ANOVA). Owing to the low degree of protein binding and the high volume of distribution of levamisole, the variations in protein binding due to different factors would not be of major clinical importance in its therapeutic application. PMID- 9203168 TI - Cardiovascular parameters of heroin-withdrawn addicts treated with guanfacine. AB - Blood pressure and heart rate were monitored in heroin addicts detoxified with guanfacine in an outpatient program. Retrospective analysis of the cardiovascular parameters from the addicts who failed this detoxification during the first 5 days of treatment (they consumed heroin within this period) showed that these patients experienced less hypotension and bradycardia when treated with the alpha 2-agonist than the group of successful patients. Therefore, the relapse to opiate use could be predicted, at least in some patients, by a minimal cardiovascular reaction to guanfacine. Opiate use was followed by significant increases in blood pressure and heart rate, as had happened in previous studies with clonidine treated addicts. This finding further confirms a similarity between alpha 2 agonists adrenergic and opiates with regard to their cardiovascular effects. PMID- 9203169 TI - Classification system of complications in neuroleptic malignant syndrome. AB - Our group treated 13 cases of neuroleptic malignant syndrome (NMS) over a period of 8 years. Based on the clinical severity of complications, the cases were classified into three types: mild, with no complications; moderate, with only respiratory disturbance; and severe, with respiratory disturbance and renal failure. The major complications affecting the prognosis of NMS are respiratory disturbance and renal failure. Renal failure is also associated with the occurrence of disseminated intravascular coagulation and rhabdomyolysis. The proposed classification system for NMS patients is useful in selecting the appropriate therapeutic strategy for this disorder. The clinical data were analyzed to determine the factors in the process of deterioration in NMS. PMID- 9203170 TI - Citicoline improves memory performance in elderly subjects. AB - Citicoline is a choline donor involved in the biosynthesis of brain phospholipids and acetylcholine extensively used in the treatment of neurodegenerative diseases. In this study we investigated the effects of the oral administration of citicoline alone (C1000:1000 mg/day; C500:500 mg/day) or in combination with nimodipine (C +NI:300 + 90 mg/day) during 4 weeks on memory performance in elderly subjects with memory deficits and without dementia (N = 24; age = 66.12 +/- 10.78 years; MMS score = 31.69 +/- 2.76). Results indicated that citicoline in comparison with placebo improves memory in free recall tasks, but not in recognition tests. A significant improvement in word recall (5.17 +/- 1.1 vs. 3.95 +/- 1.2 omissions; p < 0.005), immediate object recall (6.5 +/- 1.6 vs. 5.5 +/- 1.2 omission; p < 0.05) and delayed object recall (8.5 +/- 2.1 vs. 6.7 +/- 2.4 omissions; p < 0.005) was observed after citicoline treatment. Similar results were found in the three subgroups of treatment (8 subjects per group), suggesting that citicoline possesses memory-enhancing activity at doses of 300 1000 mg/day. A decrease in systolic blood pressure and minor changes in lymphocyte cell counting were also observed in old subjects after receiving citicoline. These effects are consistent with the vasoregulatory and neuroimmune actions of citicoline and suggest that this compound may improve memory by acting on mechanisms of brain neurotropism and cerebrovascular regulation. According to the present results, showing that citicoline improves memory performance in elderly subjects, we concluded that this molecule is suitable for the treatment of memory deficits in old people. PMID- 9203171 TI - Distribution of alpha-integrin subunits in fetal polycystic kidney diseases. AB - An alteration in cell/matrix interactions is one of the suggested mechanisms leading to cyst formation in polycystic kidney diseases. Most of these interactions are mediated by beta 1-integrins, a subfamily of integrin receptors, formed by the association of the beta 1-chain with different alpha-subunits. To date, no study on alpha-integrin subunit distribution during the early stages of cyst development has been reported. Using immunofluorescence, we analyzed the distribution of alpha-integrin subunits (alpha 1, alpha 2, alpha 3, alpha 5, and alpha 6) and basement membrane proteins in kidneys of fetuses with autosomal dominant (ADPKD) or autosomal recessive polycystic kidney disease (ARPKD). The distribution was compared with that observed in normal fetal and post-natal kidneys, and in fetal cystic dysplasia and Meckel syndrome. Marked increase in alpha 1-integrin staining was observed in normal and cystic collecting duct cells of both polycystic diseases (PKD), compared with normal and cystic controls. The distribution of integrin subunits alpha 2, alpha 3, and alpha 6 was irregular in cyst epithelial cells of PKD and cystic controls. The increased expression of the alpha 1-subunit specifically observed in PKD collecting duct cells may be an early consequence of the genetic defect in ARPKD. In ADPKD it parallels the reported expression of polycystin, the protein product of PKD1. The irregular expression of alpha 2, alpha 3, and alpha 6 integrin subunits observed in all types of cysts suggests that cell/matrix interactions are altered early and may participate in the development of cysts, perhaps by contributing to the deregulation of cell survival in cystic diseases. PMID- 9203172 TI - Effect of interleukin-8 on glomerular sulfated compounds and albuminuria. AB - To evaluate the effect of interleukin-8 (IL8) on glomerular basement membrane (GBM) sulfated compounds and albuminuria, we infused IL8 in 1% bovine serum albumin (BSA) for 5 days into the left renal artery of Holtzman male rats at the rate of 10 microliters/h using an osmotic pump. Control rats received 1% BSA. A significant increase in urinary albumin/creatinine ratio was seen on the last day of IL8 infusion (0.38 +/- 0.11, mean +/- SEM) when compared with albumin/creatinine ratio prior to infusion (0.19 +/- 0.04, P = 0.04). No significant differences in urinary albumin excretion prior to and after infusion of 1% BSA were observed. On the last day of infusion, rats were injected with 35sulfate (1.0 mCi/200 g body weight) intraperitoneally and killed after 8 h. Glomeruli were isolated and GBM obtained. After 5 days of IL8 administration, there was a significant increase in 35sulfate uptake by GBM of the infused kidney (76 +/- 10 cpm/dry glomerular weight, mean +/- SEM) compared with the uptake seen in the contralateral kidney (53 +/- 9, P = 0.05). The in vivo infusion of IL8 increased the 35sulfate uptake by GBM and augmented the urinary albumin/creatinine ratio, suggesting that IL8 may induce albuminuria by altering the metabolism of the GBM sulfated compounds. This hypothesis needs to be confirmed by studies on glomerular charge selectivity and GBM anionic sites during the course of the infusion. Moreover, the persistence of the effect needs to be evaluated by prolonging the infusion for more than 5 days. PMID- 9203173 TI - Somatic and renal effects of growth hormone in rats with chronic renal failure. AB - Recombinant human growth hormone (rhGH) has been widely used to improve growth in children with chronic renal failure (CRF). However, there has been great concern that GH may aggravate renal disease and hasten the progression to end-stage renal failure. We therefore investigated the effect of prolonged administration of rhGH at various doses on somatic growth and renal function and structure in rats with CRF, divided into four groups based on rhGH dose (vehicle, 0.4, 2.0, and 10.0 IU/day). rhGH was administered subcutaneously daily for 8 weeks. The mean growth was significantly greater in rats treated with high-dose rhGH (10.0 IU) than those treated with low-dose rhGH (P = 0.0089) or vehicle (P = 0.0011). Body weight gain increased in parallel with body length (Creatinine clearance at the end of the experiment was significantly lower in rats on high or medium-dose rhGH than those on low-dose rhGH and controls (P < 0.05). The glomerular sclerosing index was greater in rats treated with higher doses of rhGH. There were significant differences between rats treated with high-dose rhGH and controls (P = 0.0144) and also between rats on medium-dose rhGH and controls (P = 0.0065). Although there was no significant difference, rats treated with higher doses of rhGH tended to excrete more protein. Renal insulin-like growth factor-I (IGF-I) content and circulating IGF-I and IGF-II levels did not significantly differ among groups. We conclude that: (1) GH improves somatic growth failure in rats with CRF, but prolonged administration of GH dose-dependently induces deterioration in renal function and structure and (2) this effect was induced neither via circulating IGF-I and IGF-II nor by local production of IGF-I, but seems to be direct. PMID- 9203174 TI - Low molecular weight protein excretion in glomerular disease: a comparative analysis. AB - We studied 23 children with steroid-sensitive nephrotic syndrome (SSNS), 21 children with steroid-resistant types of nephrotic syndrome and 32 children with other types of nephritis. Our controls were 43 apparently healthy children. We measured the urinary excretion of N-acetyl-beta-D-glucosaminidase (NAG) and the low molecular weight (LMW) protein beta 2-microglobulin (B2M), retinol-binding protein (RBP), alpha 1-microglobulin (A1M) and urine protein 1 (UP1). Results for B2M were considered only for a urine pH greater than 6.0. Comparisons were made with urine albumin excretion, glomerular filtration rate (GFR) and tubular abnormalities in selected renal biopsy samples. We found that abnormalities of LMW protein excretion occurred in between 50% (B2M) and 88% (UP1) of all subjects. In children with SSNS, A1M (r = 0.73), UP1 (r = 0.65) and NAG (r = 0.54) excretion were significantly correlated with albumin excretion, but not RBP or B2M excretion. Increased fractional excretion of A1M, B2M and UP1 and increased plasma A1M were demonstrated in 9 children with SSNS, suggesting competition for tubular reabsorption with albumin, most marked for UP1. In the steroid-resistant nephrotic and nephritic syndromes, correlation with albumin was found for all proteins. In these subjects, RBP (r = 0.37), B2M (r = 0.42) and A1M (r = 0.28) were inversely correlated with GFR, but not UP1, NAG or albumin. We found that RBP excretion was significantly greater in the presence of severe tubular abnormalities in 11 children with recent renal biopsies, but not A1M, UP1 or NAG. We conclude that LMW proteinuria is common in children with glomerular disease, and does not necessarily imply a poor prognosis. Factors other than histologically proven tubular abnormality may account for elevated LMW protein excretion. RBP is the LMW protein most closely associated with structural abnormality and least affected by increasing albuminuria. PMID- 9203175 TI - Oligotyping for HLA-DQA, -DQB, and -DPB in idiopathic nephrotic syndrome. AB - Associations of human leukocyte antigens (HLA) with the idiopathic nephrotic syndrome (NS) have mainly been described for alleles of the HLA-DR locus. In the present study the polymorphism of HLA-DQ and -DP at the molecular level was investigated in 167 children with NS (129 steroid-sensitive) using the polymerase chain reaction and sequence-specific oligonucleotides in a French and a German cohort. HLA-DR typing was also performed by classical serology. In steroid sensitive patients we observed an increased frequency of the alleles HLA DQA1*0201 and -DQB1*0201 in both populations with relative risks ranging from 3.8 to 8.5 (Pb < 0.01 to Pb < 0.00001 after Bonferoni's correction). In contrast, the frequency of HLA-DQA1*0102 and DQB1*0602 was significantly decreased. In children with frequent relapses the HLA associations were generally more pronounced than in those with infrequent or no relapses. Applying logistic regression analysis, a nephrotic child bearing DQA1*0201 or DR7 was five times more likely to be in the steroid-sensitive group of patients than in the steroid-resistant group compared with nephrotic children not bearing one of these alleles. These HLA alleles therefore seem to be useful indicators of a steroid-sensitive frequently relapsing course of NS. No associations with DPB alleles were observed, which narrows the region genetically involved in the disease susceptibility to the DR DQ region. Steroid-resistant NS was not associated with HLA. PMID- 9203176 TI - Bartter syndrome in Costa Rica: a description of 20 cases. AB - Bartter syndrome involves an overlapping set of closely related renal tubular disorders which can be subdivided into at least three clinical phenotypes: (1) classic Bartter syndrome (2) Gitelman syndrome, and (3) a neonatal variant of Bartter syndrome. In contrast to classic Bartter syndrome and Gitelman syndrome, the neonatal variant of Bartter syndrome has both the features of renal tubular hypokalemic alkalosis as well as profound systemic manifestations. Specifically, neonatal Bartter syndrome is characterized by intrauterine polyhydramnios, premature delivery, and life-threatening episodes of fever and dehydration. Most of these infants also have severe hypercalciuria with associated nephrocalcinosis and osteopenia. Over a 22-year period, 20 Costa Rican patients with a congenital syndrome that resembles neonatal Bartter syndrome have been identified and characterized. While these patients exhibit some of the clinical characteristics previously described for neonatal Bartter syndrome, this cohort also has a set of distinct features. They are predominantly female, have a later age of diagnosis, manifest a relatively unique set of physical traits, and appear to have milder clinical disease. Given these differences, it will be important to apply the emerging molecular tools to determine whether the phenotypic variability indicates genetic heterogeneity in neonatal-onset Bartter syndrome. PMID- 9203177 TI - Autosomal recessive polycystic kidney disease: long-term outcome of neonatal survivors. AB - Autosomal recessive polycystic kidney disease causes renal and hepatic dysfunction in childhood. We describe the clinical outcome of 52 children with this diagnosis born between 1950 and 1993. Currently 23 are alive, 24 dead and 5 have been lost to follow-up; 1 has been dialysed and 7 transplanted. Life-table analysis of the patients surviving the 1st month of life revealed an actuarial renal survival of 86% at 1 year and 67% at 15 years. The probability of requiring anti-hypertensive treatment was 39% at 1 year and 60% at 15 years of age. Bleeding from gastro-oesophageal varices occurred in 8 patients at a mean age of 12.5 years, and was preceded by haematological evidence of hypersplenism in 6 of them. The study indicates a relatively good prognosis for patients with this condition who survive the neonatal period and emphasises the importance of early detection and appropriate management of systemic and portal hypertension. PMID- 9203178 TI - Proteinuria and other renal functions in Wilson's disease. AB - Renal lesions have repeatedly been described in Wilson's disease (WD). We investigated the excretion of total protein, albumin, low (LMW) and high molecular weight (HMW) proteins, N-acetyl-beta-D-glucosaminidase (NAG), and calcium, as well as creatinine clearance, in 24-h urine samples of 41 patients with WD aged 6-37 (mean 17) years who had been treated for a period of 0-15 (mean 4.5) years with D-penicillamine (900 mg/day). The amount of all protein excreted was significantly increased compared with controls, 39% of patients presenting with total proteinuria more than two standard deviations from the mean of controls. The changes in protein excretion depended on the duration of treatment. LMW proteinuria was elevated almost exclusively in the first 2 years after the start of treatment, indicating early tubular damage. This is supported by an initially high excretion of beta 2-microglobulin, NAG, and calcium. Increased excretion of HMW proteins, including albumin, persisted over longer periods, which suggests glomerular injury in some patients, possibly related to the use of D-penicillamine. Creatinine clearance remained roughly within normal limits. We propose that renal function should regularly be checked in patients with WD. PMID- 9203179 TI - Renal functional reserve in transplanted and native single kidneys of children and adults. AB - Renal functional reserve (RFR) after an oral protein load was evaluated in 36 cyclosporine-treated children following kidney transplantation (Tx), in 15 kidney donors (Don), and in 15 children with single kidneys (Nx/Ag). Glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) were determined by clearances of inulin (and creatinine) and para-aminohippurate during water diuresis. Baseline and stimulated GFR and ERPF were determined and RFR was calculated as the difference between stimulated and baseline values. Baseline GFR and ERPF in Tx were lower than in Don and Nx/Ag. Both GFR and ERPF increased significantly in all groups from baseline to stimulated values. RFR GFR was 23% +/- 3%, 20% +/- 3% and 15% +/- 3% in Tx, Don, and Nx/Ag and RFR ERPF 35% +/- 4% in Tx, which was significantly higher than 20% +/- 4% and 15% +/- 3% in the two other groups respectively. Stimulated GFR and ERPF in Tx correlated with kidney length. No differences were seen in recipient-donor pairs, except for higher fractional increases of ERPF in recipients. There was no correlation between RFR measured by clearance of creatinine and clearance of inulin. In conclusion, cyclosporine-treated children following renal Tx were found to have a renal reserve capacity. PMID- 9203180 TI - Comparison of Neoral and Sandimmun cyclosporin A pharmacokinetic profiles in young renal transplant recipients. AB - A major factor influencing whole blood cyclosporin A levels in young children with renal transplants is the variable absorption of Sandimmun (SIM). Neoral (NEO) is a new microemulsion of cyclosporin A (CYA) that has been reported to have better absorption characteristics. We compared the pharmacokinetics of SIM and NEO in nine renal transplant recipients aged less than 11 years (range 4.8 10.9 years) and observed clinical parameters during 6 months of NEO therapy. Median CYA dosage was 149 mg/m2 per day (range 98-226). We observed an increase in the maximum CYA concentration (Cmax) of 114%, an increase in area under the curve (AUC) of 71% and the time to reach Cmax was reduced from 1.75 h to 1.25 h with NEO, while 12-h trough levels (C12 h) did not change significantly. AUC correlated with C12 h for SIM (r2 = 0.833) and NEO (r2 = 0.699) and also C1.5 h for NEO (r2 = 0.775). During 24 weeks' follow-up, the coefficient of variation of CYA levels was lower for NEO (13%) than for SIM (20%). Although CYA dosages at the start and the end of 6 months on NEO were similar, only one patient was maintained on a constant dose. Four patients had acute reversible rises in plasma creatinine which responded to a 11% reduction in NEO dose; their increase in AUC was greater than those patients not showing a rise in plasma creatinine. Overall, median plasma creatinine was unchanged at the end of the study. NEO was well tolerated by the patients; temporary nausea and headache were experienced by three patients and one of them stopped NEO after 20 days. Other biochemical parameters were not significantly different on NEO. PMID- 9203182 TI - Peritonitis in continuous ambulatory peritoneal dialysis in children living in Saudi Arabia. AB - The clinical aspects of peritonitis and catheter infections were reviewed in 64 children on continuous ambulatory peritoneal dialysis living in Saudi Arabia over a period of 6 years. Peritonitis occurred in 41 children (64%). The mean time from starting dialysis to the first episode of peritonitis was 7.2 months. The incidence of peritonitis was 1 episode in 9 treatment months. Gram-negative organisms were responsible for the majority of episodes (42%), followed by Gram positive organisms (20%), and Candida albicans (6%); 32% were culture negative. Recurrent peritonitis was present in 20 cases. Catheter was replaced in 24 patients: 44% due to recurrent peritonitis. Peritoneal membrane loss occurred in 7 patients, 3 had Candida peritonitis and 3 had recurrent peritonitis due to Pseudomonas. The mortality rate was 4.6% but none of the deaths were related to peritonitis or dialysis. PMID- 9203181 TI - Effects of deflazacort immunosuppression on long-term growth and growth factors after renal transplantation. AB - Deflazacort is an oxazoline compound derived from prednisolone. We studied changes in kidney function, growth velocity, weight/height ratio, insulin-like growth factor (IGF-I), and IGF binding proteins before and after substitution of deflazacort for methylprednisone in 27 transplanted patients aged 3.1-20 years. Methylprednisone (mean +/- SEM 0.17 +/- 0.01 mg/kg per day) was replaced by deflazacort (0.29 +/- 0.01 mg/kg per day) for a period of 1-5 years. Calculated creatinine clearance did not change significantly during deflazacort treatment. Growth velocity increased from 2.6 +/- 0.5 cm/year to 5.2 +/- 0.7 cm/year (1st year) in 14 prepubertal patients. After 4 years of deflazacort treatment, height standard deviation score for chronological age did not change in 7 prepubertal patients. Mean weight/height ratio decreased by 50% (1st year) and remained reduced during follow-up. Serum IGF-I, IGF binding protein-3 (IGFBP3), IGF/IGFBP3 molar ratio, and IGF-I and II binding capacities showed no significant change; however in 5 of 6 patients IGFBP2 decreased during deflazacort therapy. Our findings suggest that immunosuppressive treatment with deflazacort is as effective as methylprednisone and may lead to an improvement in the growth prognosis of children with renal transplantation. PMID- 9203183 TI - Intermittent trimethoprim-sulfamethoxazole in children with vesicoureteral reflux. AB - The effectiveness of intermittent low-dose trimethoprim-sulfamethoxazole (TMP SMZ) for the prophylaxis of recurrent urinary infection is well established in adults. The present study assessed the effectiveness and safety of intermittent low-dose TMP-SMZ in 35 children (24 boys, 11 girls, aged 1 month to 9 years, median age 5 months) with vesicoureteral reflux; 18 children had bilateral reflux. A total of 53 refluxing ureters were graded as I in 2, II in 16, III in 19, IV in 14, and V in 2 cases. The children were given 1 mg/kg body weight of trimethoprim together with 5 mg/kg of sulfamethoxazole at bedtime every other day for 6-50 months (mean +/- SD, 22.9 +/- 11.7 months). None of the boys had a recurrence of urinary infection, while 2 of the 11 girls had a total of 7 recurrences during the prophylaxis period, with a recurrence rate of 0.027 per patient month in girls. Both girls were over 3 years and had a mildly unstable bladder. Transient neutropenia (< 1,000/microliter) developed in 2 infants during the prophylaxis period, but disappeared spontaneously. Intermittent low-dose TMP SMZ seemed very effective for the prevention of recurrent urinary infection in children with ureteral reflux even of higher grades. PMID- 9203185 TI - Urinary transforming growth factor-beta in patients with glomerular diseases. AB - We measured the urinary levels of active transforming growth factor-beta (TGF beta) by enzyme-linked immunosorbent assay in 12 healthy controls and 42 patients with various glomerular diseases, including mesangial proliferative (IgA nephritis, Henoch-Schonlein purpura nephritis, and IgA-negative mesangial proliferative glomerulonephritis) and non-proliferative (minimal change nephrotic syndrome and focal glomerulosclerosis) types. Urinary TGF-beta, expressed as a ratio to urinary creatinine (ng/mg creatinine), was elevated in patients with IgA nephritis and focal glomerulosclerosis, and was significantly higher than in patients with other types of glomerular diseases and healthy controls. There was a significant correlation between urinary TGF-beta levels and the grade of interstitial fibrosis. Among patients with proliferative-type disease, urinary TGF-beta was significantly correlated with the grade of mesangial matrix increase and the magnitude of proteinuria. The relationship between urinary TGF-beta levels and the immunostaining intensity of TGF-beta in the glomeruli was not significant. These results indicated that urinary TGF-beta reflects the grade of interstitial fibrosis in glomerular diseases and also the mesangial matrix increase in proliferative-type glomerulonephritis. Measuring TGF-beta levels in the urine might be helpful in monitoring patients with some types of glomerular disease. PMID- 9203184 TI - Dipstick only urinalysis screen for the pediatric emergency room. AB - To determine if microscopic urinalysis is needed in all pediatric emergency room patients screened for urinary tract infections (UTI), we compared the dipstick urinalysis and complete urinalysis (dipstick and microscopy) with urine cultures in 236 children, aged 3 weeks to 21 years. The ability to detect UTI by dipstick only and by complete urinalysis was the same, however microscopic evaluation added many false-positive results without detecting additional UTIs. Because the ability to detect UTI (sensitivity) is maintained, we now offer a dipstick only urinalysis to our emergency room for children 2 years of age or older, with a microscopic analysis performed automatically if dipstick results are positive. If no microscopic urinalysis is required, testing turn-around time is reduced by 12.3 min/test and the hospital charge is reduced from U.S. $32 to U.S. $12. PMID- 9203186 TI - The detection of murine autosomal recessive polycystic kidney disease using real time ultrasound. AB - Using currently available ultrasound equipment, 38 BPK and CPK mice were evaluated at 7 days of age for the presence of autosomal recessive polycystic kidney disease (ARPKD). The kidneys were less echogenic than adjacent soft tissues and measured between 5.1 and 6.3 mm from pole to pole in 32 unaffected mice and 1 with ARPKD. In 5 mice with ARPKD, the kidneys were similar in echogenicity to adjacent soft tissues and measured between 6.9 and 8.4 mm from pole to pole. Renal sonography is able to identify most mice with polycystic kidney disease prior to the development of abdominal enlargement and laboratory abnormalities, and shows promise for future applications in animal research. PMID- 9203187 TI - Humoral hypercalcemia due to an occult renal adenoma. AB - Humoral hypercalcemia refers to the elevated blood calcium levels caused by neoplasms which release a bone resorptive substance into the circulation. Previously reported infants with malignant and benign solid tumors causing humoral hypercalcemia have presented with large abdominal masses. The case we describe, a hypercalcemic infant due to an occult parathyroid hormone-related protein-containing metanephric adenoma of the kidney, shows that radionuclide bone scanning can be a useful test to identify humoral hypercalcemia. Humoral hypercalcemia stemming from a soft tissue neoplasm should be ruled out, even in the absence of clinical signs of a tumor, if bone scans show generalized uptake in the absence of hypervitaminosis D or radiological signs of bone lesions, and serum parathyroid hormone is low. PMID- 9203188 TI - Low peripheral plasma renin activity as a critical marker in pediatric hypertension. AB - In evaluating hypertensive children and adolescents, the etiological considerations should include a set of inherited disorders that share very low plasma renin activity (PRA) as a common feature. In particular among these disorders, glucocorticoid remediable aldosteronism (GRA) appears to be emerging as an important etiology of hypertension in the pediatric population. We report the evaluation of a 9-year-old Caucasian girl who presented with severe hypertension and a strong family history of early-onset hypertension. Her suppressed PRA, her family history, and her failure to respond to conventional anti-hypertensive therapy raised GRA as a potential etiology. The diagnosis was confirmed by an elevated ratio of urinary 18-oxotetrahydrocortisol to urinary tetrahydroaldosterone and genetic testing, which demonstrated the chimeric gene duplication. The molecular pathogenesis of GRA and the clinical implications are reviewed. PMID- 9203190 TI - Low-dose erythropoietin is effective and safe in children on continuous ambulatory peritoneal dialysis. AB - Hypertension is one of the most important complications of erythropoietin (rHuEPO) therapy in dialysis patients. In this study, the effect of two different dosage regiments of subcutaneous rHuEPO on blood pressure [BP] was evaluated in 20 anemic children on continuous ambulatory peritoneal dialysis (CAPD). Patients were randomized to receive rHuEPO 50 U/kg, either once a week (group 1, 50 U/kg per week) or three times a week (group 2, 150 U/kg per week). At the beginning of the study, 8 patients in group 1 and 8 patients in group 2 were on antihypertensive therapy. In group 1, the hematocrit increased gradually and significantly from 18.98% +/- 1.79% to 30.1% +/- 1.62% after 6 months, while in group 2 it rapidly increased from 19.53% +/- 1.86% to 32.4% +/- 1.11% after 3 months. A significant increase in the mean arterial BP was observed in group 2. Antihypertensive therapy had to be increased in all of the 8 previously hypertensive patients and had to be initiated in 1 of the 2 originally normotensive patients in the same group. None of the patients in group 1 required a change in antihypertensive medication. We conclude that during treatment with rHuEPO pre-existing hypertension and the dose of rHuEPO are the most important risk factors for the development or worsening of hypertension in children on CAPD, and gradual elevation of hematocrit by low-dose rHuEPO avoids the development of severe hypertension. PMID- 9203189 TI - Bilateral ureteral obstruction associated with Henoch-Schoenlein purpura. AB - Renal involvement is common in Henoch-Schoenlein purpura (HSP) and, while ureteral obstruction has been described in patients with the disease, it is rare. We report a female with HSP who developed bilateral ureteral obstruction from peri-ureteral vasculitis and subsequent ureteral ischemia. Hydronephrosis and typical findings on contrast urography indicated the diagnosis. PMID- 9203191 TI - Pasteurella multocida peritonitis in patients undergoing peritoneal dialysis. AB - A 12-year-old girl on peritoneal dialysis developed sub-clinical peritonitis due to Pasteurella multocida, following puncture of her dialysis tubing by a domestic cat. Only four other similar cases of P. multocida peritonitis have been reported in adults. This unusual form of peritonitis could be easily prevented by not allowing domestic animals to come into contact with dialysis tubings in patients undergoing peritoneal dialysis. PMID- 9203192 TI - Chronic anemia as a complication of parvovirus B19 infection in a pediatric kidney transplant patient. AB - This is a report of unexplained anemia that persisted for 4 months in an adolescent renal transplant patient receiving immunosuppression that included prednisone, tacrolimus, and mycophenolate mofetil. This patient required monthly blood transfusions for fatigue, palpitations, and hematocrit levels between 15% and 17%. In addition, his posttransplant course was notable for the development of insulin-dependent diabetes mellitus. While receiving low-dose prednisone, he was switched from tacrolimus to cyclosporin and tapered off insulin injections over the next 2 months. At 4.5 months post-transplantation, further diagnostic evaluation was suggestive of parvovirus B19 infection as the cause for our patient's chronic anemia. After testing negative for serum-specific parvovirus B19 IgM and IgG antibodies, parvovirus B19 infection was detected in blood by the polymerase chain reaction. Treatment with intravenous immunoglobulin (1 g/kg per day x 2 days) resulted in normalization of both his reticulocyte count and hematocrit within 6 weeks. At 4 months after receiving the immunoglobulin infusion, he has maintained a normal hematocrit level and stable renal function without requiring further blood transfusions. PMID- 9203193 TI - Retransplantation after post-transplant lymphoproliferative disease. AB - We present a boy who developed post-transplant lymphoproliferative disease (PTLD) 3.5 months after a first kidney transplant. The diagnosis was made after histopathological examination of the renal graft which was removed because of Pseudomonas aeruginosa septicaemia. After 2 years on dialysis, the patient received a second renal transplant. This graft continues to function after 5 years and there has been no evidence of recurrence of PTLD. This suggests that retransplantation can be undertaken in patients who have recovered from PTLD in a previous graft. PMID- 9203194 TI - Congenital cytomegalovirus infection after recurrent infection in a mother with a renal transplant. AB - A woman with a renal transplant developed a systemic cytomegalovirus infection. She recovered and 3 years later she became pregnant. She had 3 days of fever in the first trimester. She delivered an infant severely affected with congenital cytomegalovirus infection. The incidence of symptomatic congenital cytomegalovirus infection in infants born to immunosuppressed mothers who develop reactivated cytomegalovirus during their pregnancy seems high. PMID- 9203195 TI - Rhabdomyolysis and acute renal failure in a child with influenza A infection. AB - A 13-year-old previously healthy girl developed rhabdomyolysis and acute renal failure during influenza A infection. The patient recovered renal function completely with supportive therapy. This complication has been described in adult patients, but progression to acute renal failure in this context has not been reported previously in children. This diagnosis should be considered in the differential diagnosis of a pediatric patient presenting with acute renal failure and viral symptomatology. PMID- 9203196 TI - The diagnosis of renovascular disease. AB - Renovascular disease is an important cause of remediable hypertension in childhood. Specific diagnostic procedures currently available to investigate affected children include Doppler and computed duplex sonography, angiotensin converting enzyme (ACE) inhibitor sensitisation of radionuclide imaging, captopril-stimulated plasma renin activity, hypotensive responses to ACE inhibitor, renal vein renin measurements, renal angiography and magnetic resonance angiography. Carbon dioxide digital subtraction angiography and computerised tomographic and spiral angiography are also available and may play an important future role in such evaluations. Utilising this array of procedures it is usually possible to define the anatomical and functional status of the renal vasculature and be guided towards the most appropriate therapeutic manoeuvres. PMID- 9203197 TI - Molecular bases of human kidney malformations. PMID- 9203198 TI - The year 1995-1996 in review: genetically re-engineering clinical nephrology. AB - The discipline of medical genetics is rapidly transforming the face of nephrology. A number of important advances have been made during the past year in the identification of the molecular basis of renal diseases. This article will summarize how these new findings have expanded our understanding of whether diseases are homogeneous or heterogeneous entities (Bartter's syndrome versus Gitelman's syndrome), the medical basis of certain diseases of unclear etiology (enuresis), and the nature of risk factors for disease occurrence or progression (IgA nephropathy, chronic renal failure, and hemolytic uremic syndrome). It is anticipated that these exciting discoveries will become routine in the near future as medical genetics is fully incorporated into the practice of clinical nephrology. PMID- 9203199 TI - Clinical quiz. Eclampsia. PMID- 9203200 TI - Mitochondrial cytopathies and renal tubular acidosis. PMID- 9203201 TI - Neonatal Bartter's syndrome with hyperkalemia and normal urinary prostaglandin E2. PMID- 9203202 TI - Gingival hyperplasia in congenital and infantile nephrotic syndrome. PMID- 9203204 TI - Fatal Streptococcus bovis sepsis in an infant on peritoneal dialysis. PMID- 9203203 TI - Is fibrinogen degradation product-E a predictive marker for a sporadic case of hemolytic uremic syndrome with bloody diarrhea? PMID- 9203205 TI - Severe hypercalcemia, renal failure, and medullary nephrocalcinosis secondary to calcium carbonate ingestion. PMID- 9203206 TI - Does deflazacort therapy offer any advantages over conventional corticosteroids in children with renal disease? PMID- 9203207 TI - Prenatal diagnosis of haemoglobin Bart's disease by cordocentesis at 12-14 weeks' gestation. AB - Couples with alpha-thalassaemia-1 face a 25 per cent risk of having fetuses with haemoglobin (Hb) Bart's disease. Prenatal diagnosis is conventionally performed by DNA studies of chorionic villi or amniocytes obtained from chorionic villus biopsy or amniocentesis. DNA studies are expensive and time-consuming. We identified 11 affected pregnancies on abdominal ultrasound examination at 12-14 weeks when the placental thickness exceeded the mean plus 2 SD measurement for the gestational week and the cardiothoraic ratio was more than 0.5. Cordocentesis was then performed with a free hand technique. The procedures were successful in ten cases using a 26-gauge spinal needle with a 20-gauge introducer. Hb Bart's disease was confirmed in all cases by Hb electrophoresis. The procedure was unsuccessful in one case when a 22-gauge spinal needle was used. Hb study of fetal blood collected at abortion also confirmed Hb Bart's disease. In conclusion, ultrasound findings of concomitant placentomegaly and cardiomegaly at 12-14 weeks is highly specific of disease in pregnancies at risk of Hb Bart's disease. Cordocentesis and Hb study in pregnancies with these sonographic manifestations may be an alternative prenatal diagnostic approach. This diagnostic approach is of particular value in areas where resources for molecular studies are limited. PMID- 9203208 TI - Misdiagnosis of homozygous alpha-thalassaemia 1 may occur if polymerase chain reaction alone is used in prenatal diagnosis. AB - The polymerase chain reaction (PCR) is a quite sensitive diagnostic tool but its specificity may be hampered because of contamination of foreign DNA. In order to determine the diagnostic accuracy of PCR in diseases due to gross gene deletion, a total of 180 fetuses at risk of homozygous South-East Asian deletion (SEA) of alpha-globin genes were included for study. Both PCR and Southern hybridization (SH) were performed. By PCR, three of 43 affected fetuses were misdiagnosed as heterozygotes; four of 50 normal fetuses were misdiagnosed as heterozygotes; and four of 87 heterozygotes were misdiagnosed, two as normal and two as affected. Misdiagnosis in affected and normal fetuses was most likely due to maternal DNA contamination, while misdiagnosis in heterozygotes was probably due to a failed PCR. In the experiments with PCR in which we added DNA from a carrier woman to an affected or a normal fetus, a level of 1/64 and 1/16 contamination resulted in the appearance of normal and SEA breakpoint sequences, respectively. In the SH experiments using artificially contaminated DNA, a level of 1/4 contamination showed the normal and SEA bands, respectively, while a contamination level lower than 1/8 and 1/16 respectively did not reveal contamination bands. The high sensitivity of PCR makes it easier to amplify contaminated DNA. For accurate prenatal diagnosis, PCR should be performed very carefully and SH seems to be a useful back-up. PMID- 9203209 TI - First-trimester sonography of physiological midgut herniation and early diagnosis of omphalocele. AB - Embryological development was investigated by ultrasonography in a longitudinal study of 18 normal pregnancies. The appearance of midgut herniation was studied and if present, its circumference was valued. In addition, patients with an elevated risk for anomalies were scanned between 10 and 16 weeks of gestation. In 17 fetuses, the diagnosis of omphalocele was made between 11 and 16 weeks. The ratio between the circumference of the omphalocele and abdomen was calculated and the contents of the omphalocele were evaluated. Eight of the fetuses had an abnormal karyotype and a mean omphalocele/abdomen ratio of 0.38 (SD 0.191), whereas the mean ratio of omphalocele and abdomen in the eight euploid fetuses was 0.88 (SD 0.291). Regarding the contents of the omphalocele, mainly bowel loops were eviscerated in all fetuses with an abnormal karyotype. The contents of the omphalocele sac in the euploid cases contained liver in all eight fetuses. In 16 out of 17 anomalous fetuses (91 per cent), associated findings were detected. Nuchal translucency was the most frequently visualized concurrent finding and its occurrence was associated with an abnormal chromosome pattern. A small sac size is a strong indication for aneuploidy, especially in combination with nuchal translucency. Prior to 12 weeks of gestation, the diagnosis of omphalocele can be made in cases of a large omphalocele. PMID- 9203210 TI - Fetal choroid plexus cysts--is a genetic evaluation indicated? AB - A study of the association between aneuploidy and fetal choroid plexus cysts (CPCs) is presented. By reviewing the world prospective and retrospective studies, one cannot reach an agreed conclusion since different study designs were used and meta analysis is not feasible. Our experience is that as a solitary ultrasonographic finding, genetic evaluation is not indicated in cases of CPC. However, all the 'risk factors' of fetal aneuploidy such as maternal age, biochemical markers, and ultrasonographic signs may create a score by which the indication for genetic evaluation will be more sound. PMID- 9203211 TI - First-trimester urine free beta hCG, beta core, and total oestriol in pregnancies affected by Down's syndrome: implications for first-trimester screening with nuchal translucency and serum free beta hCG. AB - We have examined maternal urine concentrations of beta core, free beta human chorionic gonadotrophin (hCG), and total oestriol in 373 control pregnancies and 43 pregnancies affected by aneuploidy (including 22 cases of Down's syndrome) in an attempt to see if any of the analytes have a value in Down's syndrome screening between the tenth and 14th week of pregnancy. We have compared the performance of these analytes against nuchal translucency measurement combined with maternal serum free beta hCG at the same period of pregnancy. Our results show that levels of urine free beta hCG and beta core are increased in Down's syndrome with average multiple of the median levels of 1.81 and 2.91, respectively. Urine total oestriol was reduced (0.83) whilst maternal serum free beta hCG was increased (1.72). In trisomy 18 the levels of all analytes were reduced, although serum free beta hCG was the most discriminating. The spread of results in the control and the Down's group for urine beta core was more than three times than that for serum free beta hCG and with urine free beta hCG it was two times wider. In combination with maternal age, urine total oestriol had a 32 per cent detection rate at a fixed 5 per cent false-positive rate; urine beta core 34 per cent, urine free beta hCG 36 per cent, maternal serum free beta hCG 44 per cent, and nuchal translucency 82 per cent. In combination with nuchal translucency, urine total oestriol added an extra 1 per cent detection, urine beta core an extra 2 per cent, urine free beta hCG an extra 3 per cent, and serum free beta hCG an extra 5 per cent. It is unlikely that any of the urine markers will be of value in first-trimester screening. Optimal first-trimester screening programmes will rely for the foreseeable future on nuchal translucency, serum free beta hCG, and possibly pregnancy-associated plasma protein A. PMID- 9203212 TI - Prenatal detection of Hb mutations using transcervical cells. AB - Prenatal diagnoses were performed on six selected pairs of parents known to be carriers of Hb mutations by testing transcervical cells (TCCs) retrieved, prior to chorionic villus sampling (CVS), by aspiration of the cervical mucus from the pregnant mothers at 10-12 weeks of gestation. A concordance between the results of testing chorionic villus cells and isolated clumps of trophoblastic cellular elements was observed in four of the six cases. PMID- 9203213 TI - The role of reduced ear size in the prenatal detection of chromosomal abnormalities. AB - A prospective ultrasound study was performed between 18 and 38 weeks' gestation on 29 fetuses in a high-risk population, defined by the presence of structural anomalies, in order to investigate the usefulness of fetal ear measurements in the prenatal prediction of chromosomal abnormality. The prevalence of abnormal chromosomes was 34 per cent. The sensitivity (SE), specificity (SC), positive predictive values (PPV), and negative predictive values (NPV) of ear length for the detection of chromosomal abnormality were 80, 84.2, 72.7 and 88.9 per cent. The SE, SC, PPV, and NPV of ear width were 40, 94.7, 80 and 75 per cent. Fetal ear measurements may be a useful adjunct to the various ultrasound parameters in the prenatal detection of chromosome abnormality in a high-risk population of fetuses with structural anomaly(ies). PMID- 9203214 TI - Biochemical and morphological expression of early prenatal caprine beta mannosidosis. AB - Lysosomal storage diseases associated with early-onset pathological changes may require prenatal therapy to avert the profound effects of the metabolic error on organs, especially the central nervous system. The present investigation determined the extent of expression of beta-mannosidase deficiency in the caprine fetus at 62 days of gestation, near the end of the period of immunotolerance when donor cells can engraft in various organs without immune rejection and supply missing enzyme. Three pairs of obligate carrier goats from the beta-mannosidosis colony were mated. Out of six fetuses delivered at 62 days of gestation, one (V385) was identified by measurement of beta-mannosidase activity as the only fetus affected with beta-mannosidosis. Thin-layer chromatography and quantitation of oligosaccharides revealed the presence of tri- and disaccharides, typical of beta-mannosidosis, only in V385. Morphological analysis revealed cytoplasmic vacuolation typical of beta-mannosidosis in V385; in thyroid, spinal cord, and kidney, the pattern of vacuolation was similar to, but less severe than, that observed previously in newborn affected goats. On the basis of these results, it will be possible to determine the effects of prenatal cell transplantation therapeutic strategies performed during the period of immunotolerance by monitoring phenotypic characteristics after treatment. PMID- 9203215 TI - Strategies and outcomes of prenatal diagnosis for osteogenesis imperfecta: a review of biochemical and molecular studies completed in 129 pregnancies. AB - We completed prenatal diagnostic studies from 129 pregnancies at risk for osteogenesis imperfecta (OI). Studies in 107 pregnancies were completed by analysis of collagen synthesized by cells cultured from chorionic villus biopsies and the remaining 22 used direct mutation identification or analysis of polymorphic restriction sites in the COL1A1 gene of type I collagen. The vast majority of studies (n = 113) were obtained to identify fetuses with OI type II (the perinatal lethal form) and some fetuses affected with OI type III or IV (the deforming varieties). Of the 50 couples who had had one previous affected pregnancy with the lethal form of OI, one had a second affected pregnancy, a rate of 2 per cent. Two of the seven unaffected couples (28 per cent) who had had two previous affected pregnancies with OI type II had a third affected pregnancy; none of the three with two previous pregnancies with OI type III had a third. Pregnancies at risk for OI type I could not be ascertained reliably by biochemical analysis of cultured CVS cells but were identified by direct analysis of the causative mutation or the use of linked markers in families. All prenatal diagnostic studies were undertaken only after earlier diagnostic studies (biochemical or molecular) had been completed on the proband, a necessary strategy for accurate results. In all pregnancies at risk for OI type II, OI type III, and OI type IV studied with biochemical strategies and in pregnancies at risk for OI type I studied with molecular techniques, there were neither false negative nor false-positive results. Diagnostic information can be obtained within 20-30 days of biopsy using biochemical techniques and within 10-14 days when molecular strategies are used. PMID- 9203216 TI - Prenatal diagnosis of a deletion of 18q in a fetus associated with multiple marker screen positive results. AB - We report here the observations of positive maternal serum screening tests for Down syndrome, cytogenetic and molecular analysis, and dysmorphic fetal features in a pregnancy with 18q-syndrome. A 33-year-old primigravida was referred for genetic counselling because of multiple-marker screen positive results. At 14 weeks' gestation, the woman had a Down syndrome risk of 1:107 calculated from a maternal serum alpha-fetoprotein (MSAFP) level of 1.49 multiples of the median (MOM), a total human chorionic gonadotrophin (hCG) level of 2.42 MOM, and a serum unconjugated oestriol (uE3) level of 0.55 MOM. At 17 weeks' gestation, a repeat test showed a Down syndrome risk of 1:10 calculated from an MSAFP level of 1.09 MOM and a free beta-hCG level of 12.3 MOM. Genetic amniocentesis revealed a de novo deletion of 18q22.2-qter. Intrauterine fetal death occurred at 21 weeks' gestation. At birth, the fetus manifested clinical findings of the 18q-syndrome. The phenotype was correlated with the extent of the deletion. Linkage analysis of the family confirmed the extent and paternal origin of the deletion. PMID- 9203217 TI - Prenatal diagnosis of laryngeal atresia in two cases of congenital high airway obstruction syndrome (CHAOS). AB - Complete occlusion of the upper airways is known to cause secondary morphological changes, including bilaterally enlarged hyperechogenic lungs, dilated trachea, and hydrops. Prenatal diagnosis of upper airway obstruction has been described in several cases. In these reports, the diagnosis was primarily attributed to indirect signs, and the authors were in doubt as to whether the location of the obstructed area (larynx, glottis or trachea) could be precisely visualized by ultrasound. In this paper two cases are reported presenting the features of congenital high airway obstruction syndrome (CHAOS) diagnosed at the 22nd week of gestation. In both cases, the upper neck was perfectly visualized in a coronal plane. At the onset of fetal breathing movements, the stenotic larynx remained in a closed position. By positioning a colour signal on the fluid-filled dilated trachea, we noticed absence of flow throughout the onset of breathing activity. We conclude that the atretic area must be situated at the level of the larynx. Because of the poor prognosis of laryngeal stenosis and the presence of associated anomalies, both pregnancies were terminated in the 23rd week. This report shows that the application of colour and spectral Doppler may be helpful in the differential diagnosis of fetuses with CHAOS. PMID- 9203219 TI - Yet another variation on the theme of chromosome 18 heteromorphisms? PMID- 9203218 TI - Diagnostic echographic findings in cryptophthalmos syndrome (Fraser syndrome). AB - We report two male siblings with cryptophthalmos syndrome (Fraser syndrome), an autosomal recessive multiple malformation syndrome with cryptophthalmos, abnormal genitalia, and syndactyly of the fingers and toes as major clinical symptoms. Renal anomalies (uni- or bilateral agenesis) occur in 85 per cent. In the second trimester of both pregnancies (at 23.5 and 18.5 weeks, respectively), echographic examination revealed multiple anomalies: oligoamnios sequence and fetal hydrops with nuchal oedema. Contrasting with the oligohydramnios, the lungs were voluminous and hyperechogenic. Fetopathological examination revealed that the oligoamnios sequence was due to major renal abnormalities (bilateral renal agenesis in the first, and unilateral renal agenesis and contralateral multicystic renal dysplasia in the second sibling). Laryngeal substenosis had resulted in another malformation sequence consisting of overdistended lungs, and non-immune fetal hydrops. The present experience shows that in the presence of an oligoamnios sequence with contrastingly voluminous, hyperechogenic lungs, the diagnosis of cryptophthalmos syndrome should seriously be considered in the differential diagnosis. PMID- 9203220 TI - Confirmation of fetal aneuploidy with primed in situ hybridization on amniotic fluid during selective fetal reduction. PMID- 9203221 TI - Turner's syndrome 45,X found by coelocentesis. PMID- 9203223 TI - Current awareness in prenatal diagnosis. PMID- 9203222 TI - Isochromosome 18q revisited. PMID- 9203224 TI - Growth hormone response to the GABAB agonist baclofen in major depressive disorder. AB - Growth hormone (GH) response to the gamma-aminobutyric acid B receptor agonist, baclofen, was measured in 16 male patients with major depressive disorder and in 16 age-matched healthy male controls. No significant differences were found in the GH response to baclofen between the depressed patients and controls. On repeat testing, the GH response to baclofen showed significant retest reliability in both groups. There was no significant correlation between serum baclofen levels and the GH response to baclofen. Age significantly correlated with GH response, with older subjects having lower GH response to baclofen. These data do not suggest that a blunted GH response to baclofen. represents a specific neuroendocrine feature of major depression. PMID- 9203225 TI - Dexamethasone suppression of the effects of cocaine on adrenocortical secretion in Lewis and Fischer rats. AB - There is evidence to suggest that cocaine acts centrally to enhance adrenocortical secretory activity and this effect may be associated with the reinforcing properties of this drug. Lewis (LEW) and Fischer (F344) rats are inbred strains which differ in their responses to the reinforcing effects of cocaine. Previous findings from this laboratory have demonstrated differences in the hypothalamic-pituitary-adrenocortical (HPA) responses to cocaine between these strains. To determine whether strain differences in glucocorticoid responsiveness play a role in the differential effects of cocaine on plasma corticosterone (CS) secretion in these strains, experiments were designed to suppress the HPA response to cocaine in these two rat strains. HPA activity was attenuated by central administration of the glucocorticoid agonist dexamethasone (DEX) using osmotic minipumps. A constant infusion of artificial cerebrospinal fluid or DEX (50, 100 or 500 ng/h) was delivered into the lateral ventricle of LEW and F344 rats. Four days later, the rats were challenged with cocaine HCl (0, 20 and 40 mg/kg, i.p.), and the plasma CS response 15 min later was quantified. Cocaine-induced alterations in circulating plasma CS were reduced in a dose related manner by centrally administered DEX in both strains. Significant strain differences in the effects of DEX on the plasma CS response to cocaine were observed, suggesting that LEW rats were more sensitive to DEX suppression of HPA activity than F344 rats. DEX also produced dose-related effects on body weight in both strains and decreased adrenal weight at the highest dose in F344 rats. Blood collected on the final day of the experiment demonstrated that central infusions of DEX decreased plasma ACTH concentrations in both strains compared to control rats. These studies indicate that central administration of DEX produces a feedback inhibition of cocaine-induced glucocorticoid release and that LEW rats are more sensitive to DEX suppression than F344 rats. PMID- 9203226 TI - Behavioural and physiological consequences of a single social defeat in Roman high- and low-avoidance rats. AB - The behavioural and physiological consequences of a single social defeat were studied in Roman high-avoidance (RHA) and Roman low-avoidance (RLA) rats, two rat lines with a genetically determined difference in the way of responding to or coping with stressors. Animals were subjected to social defeat by placing them in the cage of an aggressive male conspecific for 1 h. In both RHA and RLA rats, social defeat induced a profound increase in body temperature during the circadian resting phase, lasting for up to 10 days after the conflict. The increase in resting temperature was paralleled by a slight decrease in spontaneous home cage activity. Food intake and growth were suppressed for a number of days, resulting in a long-lasting lower body weight compared to non stressed control animals. An open field test 2 days after defeat showed a social stress-induced decrease in locomotion in a novel environment. Despite the well known differentiation between RHA and RLA rats in their behavioural and neuroendocrine response pattern to acute environmental challenges, the present study did not show major differences in the long-term consequences of social defeat. PMID- 9203227 TI - Cortisol feedback effects on plasma corticotropin levels in healthy subjects. AB - An abnormality of rapid cortisol feedback on activity of the hypothalamic pituitary-adrenal axis has been reported in depression. However, there is controversy regarding the existence of rapid cortisol feedback on corticotropin (ACTH) secretion in humans. We investigated the effects of cortisol on ACTH levels in healthy subjects using a placebo-controlled, double blind, random assignment, cross-over design. Ten medication-free volunteers with no psychiatric history and no active medical problems underwent a standard protocol on two occasions separated by at least 2 weeks. Each time, subjects were admitted to a General Clinical Research Center and had infusion of 15 mg cortisol (hydrocortisone sodium succinate) over 120 min or placebo. Serum levels of cortisol and plasma ACTH levels were determined at baseline and over the 4 h after the start of the infusion. Over the two GCRC admissions subjects received both cortisol and placebo infusions, and the order of the two infusions was randomized. Compared to placebo, cortisol infusion produced a significant decrease in plasma ACTH levels beginning within 60 min from the start of the infusion. We conclude that cortisol infusion produces early inhibition of ACTH secretion in normal humans. PMID- 9203228 TI - Responses to alpha 2-adrenoceptor blockade by idazoxan in healthy male and female volunteers. AB - Seven male and five female volunteers underwent double-blind infusions of the alpha 2-adrenoceptor antagonist idazoxan (100 and 200 micrograms/mg) and placebo in random order. Blood pressure, plasma norepinephrine, growth hormone and subjective responses were measured. The higher dose of idazoxan produced increases in blood pressure, norepinephrine and growth hormone and slight increases in anxiety. Both subject age and sex appeared to influence the magnitude of responses. PMID- 9203229 TI - A meta-analysis of the effect of hormone replacement therapy upon depressed mood. AB - This meta-analysis had two objectives: (a) to aggregate data from studies that used hormone replacement therapy (HRT) and a quantitative measure of depressed mood in order to examine the effectiveness of HRT upon menopausal depressed mood; and (b) to review the methodologies of this literature base. The overall effect size for HRT was 0.68. This indicated that the average treatment patient had lower levels of depressed mood than 76% of the control patients. Analyses of specific hormone treatments suggested that (a) estrogen significantly reduced depressed mood (ES = 0.69); (b) progesterone alone, and in combination with estrogen, was associated with smaller reductions in depressed mood (ES = 0.39, ES = 0.45, respectively); and (c) androgen alone and in combination with estrogen was associated with greater reductions in depressed mood (ES = 1.37; ES = 0.90, respectively). In summary, HRT appeared to be effective in reducing depressed mood among menopausal women. The methodological review indicated that most studies used adequate sample sizes, controlled research designs, random assignment, double-blind treatment manipulations, and valid and reliable measures of depression. Limitations in the interpretation of these results are discussed and recommendations for improved methodology are provided. PMID- 9203231 TI - Repeated REM sleep deprivation after chronic haloperidol administration in the rat. AB - Repeated haloperidol administration produces up-regulation of dopamine (DA) receptors. REM sleep deprivation (REMSD) does also, but in addition, has been shown to produce REM sleep rebound. Should DA receptor up-regulation play a role in REM sleep rebound, haloperidol could conceivably have effects similar to those observed following REMSD. This is the central question investigated in this study. Male Wistar rats were prepared for sleep recordings. They were randomly assigned to the following groups: group 1, REMSD by small platforms (40 h REMSD + 8 h recording); group 2, was the large platform control group (40 h in large platforms + 8 h of recording); group 3, received 2-week daily administration of haloperidol (3 mg/kg, i.p.) plus REMSD (40 h REMSD + 8 h of recording); group 4, 2-week administration of haloperidol (3 mg/kg) without sleep manipulation and at the end 40 h were allowed to elapse, following which 8 h of sleep recordings was carried out. In each group the sleep manipulation and/or sleep recordings were repeated five consecutive times. Repeated REMSD produced increases of REM sleep time after each recovery in group 1. Large platforms did not produce increases of REM sleep during the recovery trials. The 2-week administration of haloperidol plus REMSD prevented REM sleep rebound (group 3). The 2-week administration of haloperidol without sleep manipulation (group 4) produced a REM sleep reduction. Dopamine modulation seems not to be important for REM sleep rebound. Hypersensitivity of DA receptors developed after REMSD may be an epiphenomenon associated with this sleep manipulation, but seems not to participate in REM sleep enhancement after REMSD. PMID- 9203232 TI - The continuous and simultaneous blood flow velocity measurement of four cerebral vessels and a peripheral vessel during cigarette smoking. AB - There has been no consensus about the acute effect of cigarette smoking on cerebral blood flow, and the continuous change of flow in four cerebral vessel flow with peripheral flow during different kinds of cigarette smoking has not been reported until now. Our results indicate smoking increases the flow of four cerebral vessels almost at the same time and with the same pattern. Many cerebral vessels began to show increases about 10 s after commencement. In most cases, cerebral blood velocity began to decrease between 10 and 20 s after cessation. Blood flow in peripheral vessels decreases after commencement, which is thought to be the effect of nicotine. The effect of high nicotine cigarettes is greater than that of low nicotine cigarettes. Continuous and simultaneous measurement of cerebral vessels by ultrasonic Doppler is though to be the only way to establish the detailed blood flow changes during smoking. PMID- 9203230 TI - Evidence of acoustic startle hyperreflexia in recently detoxified early onset male alcoholics: modulation by yohimbine and m-chlorophenylpiperazine (mCPP). AB - Preclinical studies suggest that acoustic startle amplitude is increased during ethanol withdrawal. The current study evaluated the effects of intravenous infusion of the alpha 2-adrenergic antagonist, yohimbine (0.4 mg/kg), the serotonin partial agonist m-chlorophenylpiperazine (mCPP, 0.1 mg/kg), and placebo administered to 22 male patients meeting DSM-III-R criteria for alcohol dependence and 13 male healthy subjects. Patients and healthy subjects completed 3 test days under double-blind conditions in a randomized order. Patients were sober for 12-26 days prior to testing. On each test day, participants completed startle testing 80 min following drug infusion. Stimuli with varying intensities (90, 96, 102, 108, 114 dB) were presented in a randomized order balanced across four blocks. Stimuli consisted of 40-ms bursts of white noise administered every 45-60 s for 15-20 min through headphones. Analyses indicated that patients exhibited elevated acoustic startle magnitudes on the placebo day relative to healthy subjects. In patients, the magnitude of startle amplitudes elicited at 90 dB, but not 114 dB, correlated significantly with the number of previous alcohol detoxifications. Yohimbine increased startle magnitudes and reduced startle latencies relative to placebo and mCPP in both patients and healthy subjects. mCPP did not alter startle magnitude in either group. Yohimbine also increased the probability that a 90-dB stimulus produced a startle response in healthy subjects, but not in patients. Blunting of yohimbine effects on startle probability may reflect the baseline elevations in startle probability levels in patients, but may also be consistent with other evidence of reduced postsynaptic, but not presynaptic, noradrenergic function in these same patients. These data replicate and extend previous reports indicating that yohimbine facilitates the acoustic startle response in humans. They also further implicate the number of episodes of ethanol withdrawal as a factor influencing subsequent neurobiological responsivity in chronic alcoholic patients. Based on the current data, future research should explore whether measurement of the acoustic startle response provides an objective quantitative severity measure of ethanol withdrawal. PMID- 9203233 TI - Effects of naltrexone on amphetamine-induced locomotion and rearing: acute and repeated injections. AB - The present experiment investigated the ability of the opiate receptor antagonist naltrexone to block the increased locomotion and rearing produced acutely by amphetamine as well as the sensitization of these responses produced when this drug is administered repeatedly. Rats in different groups received an injection of amphetamine (1.5 mg/kg, i.p.) or saline preceded 30 min earlier by an injection of naltrexone (0, 0.5, 1.0, 5.0 or 10.0 mg/kg, i.p.). Naltrexone dose dependently reduced the rearing but had no effect on the locomotion produced by this dose of amphetamine. The locomotion and rearing observed following saline were not affected. This pattern of results was observed following each of six additional pairs of injections, one pair of injections given every third day. Once, soon (2-4 days) and once, long (9-12 days) after the last injection, all animals were injected with amphetamine (0.75 mg/kg, i.p.) in the absence of naltrexone (tests for sensitization). Animals having been pre-exposed to amphetamine preceded by naltrexone showed no evidence of sensitized rearing on either test, indicating that naltrexone blocked sensitization of this response to amphetamine. These animals, however, exhibited sensitized locomotion on both tests. These results suggest an important but complex role for dopamine-opioid interactions not only in the production of acute locomotor responding to amphetamine but also in the sensitization of locomotor responding when this drug is administered repeatedly. The present findings also suggest that amphetamine induced rearing is more dependent than locomotion on neuronal mechanisms involving dopamine-opioid interactions. PMID- 9203234 TI - A clinical evaluation of risperidone in the treatment of schizophrenia: a 10 week, open-label, multicenter trial. ARCS Study Group. Assessment of Risperdal in a Clinical Setting. AB - The efficacy and safety of risperidone have previously been demonstrated in controlled clinical trials in hospitalized chronic schizophrenia patients who met strict research criteria. The present study was designed to evaluate the efficacy and safety of risperidone in a heterogeneous patient population. Patients were enrolled in the study if they had a diagnosis of schizophrenia (DSM-III-R) with or without acute exacerbation. Of the 945 patients from 158 psychiatric centers who entered this phase IV study, 558 completed the 10-week trial. During week 1, the dose of risperidone was titrated to 6 mg/day, maintained there for 1 week, and then adjusted over a 4-week period as clinically necessary; the dose was then fixed for the final 4-week period. The mean dose of risperidone at endpoint was 5.9 mg/day. Patients were evaluated at baseline and at weeks 2, 6, and 10, using Clinical Global Impression scale, Psychotic Symptoms Assessment scale, and Global Assessment of Functioning scale. Significant improvement in mean scores was found on each of these measures at endpoint. Comparable results were obtained at week 10 in treatment-resistant and non-treatment-resistant patients. Risperidone was generally well tolerated and the severity of extrapyramidal symptoms was significantly reduced at endpoint. PMID- 9203235 TI - Chronic exposure to cadmium attenuates behavioral sensitization to morphine. AB - The purpose of this investigation was to assess the impact of dietary cadmium on morphine-induced changes in locomotor activity. Adult male rats were exposed ad libitum to an adulterated food supply containing 100 ppm added cadmium chloride, or an identical diet containing no added cadmium, for 45 days prior to testing for the locomotor activating effects of successive daily morphine administration (0, 5, 10, or 20 mg/kg per session) on locomotor activity. On day 1 of testing, increasing doses of morphine produced a dose-related suppression of activity, and this sedative effect was greater in control than in cadmium-exposed animals. Repeated morphine administration resulted in tolerance to the sedative effects of the drug, and a systematic elevation of locomotor activity over the 14-day testing period was observed, with the augmentation (sensitization) effect more pronounced in control than cadmium-exposed animals. There was no indication that conditioning (context) events played a role in the effects observed here. PMID- 9203237 TI - CP-94, 253: a selective serotonin1B (5-HT1B) agonist that promotes satiety. AB - The selective 5-HT1B agonist CP-94,253 (3- (1,2,5,6-tetrahydro-4-pyridyl)-5 propoxypyrrolo[3, 2-b] pyridine) (5-40 mumol/kg) reduced the intake of both pellets and a 10% solution of sucrose (ID50 = 12.5 and 22.8 mumol/kg, respectively) in mildly deprived rats. Time-sampled observations revealed that CP 94,253 terminated feeding earlier, without disrupting the continuity of feeding. CP-94,253 increased standing but did not promote resting during satiation. Microstructural analysis of licking indicated that CP-94,253 decreased the frequency, but not the size, of bursts and clusters of licks without altering oral motor efficiency. The peripherally acting 5-HT1B agonist, CP-93,129 (3 (1,2,5,6-tetrahydropyrid-4-yl)pyrrolo[3,2-b]pyrid-5-one) had no effect on food intake. These results imply that CP-94,253 probes a role for central 5-HT1B receptors in the regulation of meal size and duration, but that recruitment of other 5-HT receptor subtypes may be needed for the full expression of satiety. PMID- 9203236 TI - Anxiolytic-like effects of PNU-101017, a partial agonist at the benzodiazepine receptor. AB - PNU-101017 is a chemically novel ligand at the benzodiazepine recognition site of cloned GABAA receptors. It was reported to potentiate GABA-mediated chloride current in cultured cells with a moderate intrinsic activity and a biphasic dose response relationship. In this study, we confirmed that PNU-101017 has a partial agonist-like effect in the antagonism of metrazole-induced seizures in mice. It produced no sedation or ataxia, but did antagonize diazepam-induced motor deficit of mice in the rotarod test. PNU-101017 was weakly active in anti-conflict anxiolytic tests, but attenuated the plasma corticosteroid response to mild stress in rats. It also antagonized stress-induced elevation of cerebellar cGMP levels in mice. Like chlordiazepoxide, it increased drinking of saline solution in thirsty rats. PNU-101017 did not potentiate the CNS-depressant effects of ethanol, and produced no evidence of physical dependence when administered repeatedly. Agonists with low intrinsic activity at the benzodiazepine receptor, such as PNU-101017, should be further explored for therapeutic uses. PMID- 9203238 TI - Baclofen suppression of cocaine self-administration: demonstration using a discrete trials procedure. AB - We have previously reported that rats display a circadian pattern of cocaine self administration if access to drug is limited to 10-min discrete trials that are separated by at least 20 min. In the present study, the pattern of cocaine intake (1.5 mg/kg per injection) was studied in two large groups of animals that were maintained on different 12-h light/dark cycles (3 a.m. to 3 p.m. versus 10 a.m. to 10 p.m.). Regardless of the time of light onset, a circadian pattern of cocaine self-administration was observed. Maximum cocaine intake occurred during the final 6 h of the dark period and was followed by a relative abstinence period during the light phase. This highly predictable pattern of drug taking behavior provided an opportunity to explore the effect of baclofen, a GABAB agonist, on the initiation of self-administration behavior. In two separate studies, acute treatment with baclofen (1.25-5.0 mg/kg) was shown to suppress cocaine intake for at least 4 h. Baclofen had no significant effect on responding for food reinforcement. Previous results have indicated that baclofen appears to reduce specifically the motivation to respond for cocaine. Together, these data suggest that baclofen should be considered as a possible pharmacotherapeutic agent in cocaine addiction. PMID- 9203239 TI - Midazolam-induced rapid changes in licking behaviour: evidence for involvement of endogenous opioid peptides. AB - The role of endogenous opioid peptides in the effects of midazolam on ingestive behaviour was investigated using a detailed analysis of licking behaviour in the rat. Midazolam (1.8 mg/kg i.p.) was administered in combination with either flumazenil (10 and 20 mg/kg i.p.) or naloxone (0.1 and 0.3 mg/kg i.p.). The effect on licking patterns during 60-s exposures to a range of concentrations of a fat emulsion (Intralipid) was then recorded. Midazolam significantly increased the total number of licks for Intralipid by increasing the mean bout duration. This effect is consistent with the proposal that benzodiazepines enhance palatability. Flumazenil and naloxone were ineffective when administered alone, but both drugs blocked the effect of midazolam on total number of licks by selectively attenuating mean bout duration. Midazolam also produced a significant decrease in the intrabout lick rate, probably due to the muscle relaxant effects of this drug. This decrease in the intrabout lick rate was reversed by pretreatment with flumazenil but not by naloxone. The results suggest that endogenous opioids may be important for the palatability effects of midazolam, but may not be involved in the muscle relaxant effects of this drug. PMID- 9203240 TI - Increased aggression after ethanol self-administration in male resident rats. AB - In order to study experimental alcohol intake that leads to heightened aggression, we established ethanol self-administration in aggressive rats. The focus was on low doses of self-administered ethanol and to assess their effects on aggressive behavior in resident rats, using a limited access paradigm followed by a 5-min confrontation with an intruder. In the first phase of the experiment, rats were established as "residents", and their consistent aggressive behavior in confrontations with an intruder was verified. In the second phase, these resident rats were trained to self-administer alcohol, using a sucrose-fading technique. In the third phase, alcohol self-administration was followed by intruder confrontations in order to study the effect of alcohol on aggression. Confrontations after ethanol consumption leading to low (5-20 mg/dl) and moderate (20-50 mg/dl) blood alcohol concentration (BAC) were compared to confrontations without alcohol, each animal serving as its own control. On average, the group showed no change in aggressive behavior after low or moderate ethanol intake. However, six out of 16 individuals significantly increased the number of attack bites and the duration of aggressive behavior by up to 90% after alcohol self administration. When these rats were assigned post-hoc to an alcohol heightened aggression group, the group was characterized by a 40% increase in number of attack bites and a 90% increase in aggressive posture over control (BAC 0 mg/dl), whereas the alcohol non-heightened aggression group showed no significant changes. These results extend previous observations of increased aggression in a subpopulation of animals after experimenter-administered ethanol in mice, rats and monkeys to self-administered alcohol. Using this animal model, individuals showing enhanced or reduced aggression after oral alcohol self-administration can be characterized behaviorally, physiologically, and neurochemically. PMID- 9203241 TI - Behavioural and neurochemical characteristics of phentermine and fenfluramine administered separately and as a mixture in rats. AB - Clinical case studies suggest that combined administration of the serotonergic agent fenfluramine (FEN) and the weak amphetamine-like anorexic agent phentermine (PHEN) may be useful in the treatment of alcohol and cocaine addictions. The present experiment examined the nature of the interaction between the two agonists using the drug discrimination paradigm. In vivo microdialysis served to examine the neurochemical profile of dopamine and serotonin release in the nucleus accumbens. In conscious rats, acute injections of FEN (1.0-2.0 mg/kg i.p.) or PHEN (1.0-2.0 mg/kg i.p.) selectively elevated levels of serotonin and dopamine in the nucleus accumbens, respectively. A mixture (1 mg/kg of each) increased levels of both amines by similar magnitudes to those observed with each individually. Three groups of Sprague-Dawley rats were trained to discriminate (1) FEN (1.0 mg/kg i.p.) alone, (2) PHEN (1.0 mg/kg i.p.) alone or a mixture (3) PHEN+FEN (1 mg/kg of each, i.p.) from saline under a fixed ratio (FR-10) schedule of food reinforcement. Rats acquired the mixture discrimination rapidly, while for the other groups the training dose had to be increased to 2.0 mg/kg to attain stimulus control. The individual components of the mixture at the training dose generalized partially to the mixture, and complete generalisation was observed following 3.0 mg/kg FEN or PHEN. Rats trained to discriminate the individual components showed respective cross-generalisation profiles. Generalisation to cocaine (0.3-10.0 mg/kg i.p.), amphetamine (0.1-3.0 mg/kg i.p.) and nicotine (0.1 0.8 mg/kg s.c.) was greatest in the MIX-trained rats, while partial or no generalisation was observed in rats trained to discriminate the individual compounds. From the present results, it may be concluded that the two drugs given as a mixture do not produce a novel cue. Rather, these aminergics appear to interact additively. Furthermore, the dual stimulation of the amines by the mixture may be the basis for the cueing effects of the FEN+PHEN drug mixture, and its effectiveness in treating drug addictions. PMID- 9203244 TI - Sudden, unexpected death in epilepsy in children. AB - Sudden, unexpected death in epilepsy (SUDEP) remains a controversial and enigmatic syndrome, particularly in children where the incidence, prevalence and risk factors may, and probably do, differ from adults. This study demonstrates (and further reinforces) the difficulties and inability of retrospective and coroner/death certificate-derived data in identifying the frequency of SUDEP in children. PMID- 9203242 TI - Discriminative stimulus properties of cocaine: enhancement by beta-adrenergic receptor antagonists. AB - Although many of the behavioral effects of cocaine are widely believed to be mediated by blockade of dopamine transporters, recent studies suggest that norepinephrine (NE) may play a modulatory role. In this study, selective and nonselective beta-adrenergic receptor antagonists were administered alone or in combination with cocaine (2.5 mg/kg, i.p.) to rats trained to discriminate a low dose (2.5 mg/kg) from a high dose of cocaine (10 mg/kg) in a two-lever, FR10 drug discrimination procedure. The central beta 2/beta 1-adrenergic antagonists (-) propranolol and tertatolol, and the beta 2-adrenergic antagonist, ICI 118,551, produced high-dose appropriate responding in a dose-related manner when administered (i.p.) in combination with the low training dose of cocaine. In contrast, neither the peripheral beta 2/beta 1-adrenergic antagonist, nadolol, nor the central beta 1-adrenergic antagonist, beta-xolol enhanced the behavioral effects of the low dose of cocaine in a manner comparable with that produced by compounds with central beta 2-adrenergic antagonist properties. Also in contrast to findings obtained using beta-adrenergic antagonists, neither the alpha 1 adrenergic agonist cirazoline, nor the alpha 2-adrenergic ligands (+/-)-efaroxan and UK-14304 enhanced the behavioral effects of the low dose of cocaine. Overall, these results suggest that central beta 2-adrenergic receptors may play a modulatory role in the discriminative stimulus effects of cocaine. PMID- 9203243 TI - Established antiepileptic drugs. AB - Despite the recent entry into the market-place of a range of new pharmacological treatments for epilepsy, most patients still receive the standard antiepileptic drugs. This review considers the clinical place and practical use of these agents. Detailed consideration is given to carbamazepine, phenytoin, sodium valproate, phenobarbital and ethosuximide, with lesser emphasis on primidone, clobazam and clonazepam. Individualization of therapy, polypharmacy, refractory epilepsy, therapeutic drug monitoring, pregnancy, withdrawing treatment, epilepsy prophylaxis and referral to an epilepsy centre are also discussed. The paper concludes with a statement of 12 basic rules in prescribing established antiepileptic drugs. PMID- 9203245 TI - Lack of pharmacokinetic interaction between remacemide hydrochloride and sodium valproate in epileptic patients. AB - A randomized, double-blind, placebo-controlled cross-over study of adjuvant treatment with remacemide hydrochloride was carried out in 17 patients taking sodium valproate (VPA) as monotherapy. Plasma concentration profiles of VPA, remacemide, and its active desglycinyl metabolite (ARL12495XX) were determined following single (300 mg) and multiple dosing (150 or 300 mg twice daily) of remacemide hydrochloride for 14 days with a 300-mg final dose. Central nervous system side-effects were more common at the higher dose, which prompted dosage reduction to 150 mg twice daily for subsequent patients partway through the study. The mean area under the concentration-time curve, peak concentration and pre-dose concentration of VPA were unchanged by remacemide hydrochloride in three patients on the higher and in 10 patients on the lower dose of remacemide. The pharmacokinetic parameters of remacemide and its active metabolite in the VPA treated patients were similar to those described previously in healthy volunteers. Thus, remacemide hydrochloride does not interfere with the pharmacokinetics of VPA and vice versa. PMID- 9203246 TI - An audit of the organization of adult epilepsy services in the UK: a comparative review of epilepsy and general neurology clinics. AB - A survey of all consultant neurologists was carried out to investigate the current provision of hospital-based adult epilepsy services in the UK and to compare the level of services offered by epilepsy and general neurology clinics. The valid response rate was 75%. Fifty-four epilepsy clinics were identified led by 43 neurologists in 46 hospitals. Over half the major neurological centres represented in the dataset had epilepsy clinics (31/58). Epilepsy clinics were significantly more likely than general neurology clinics to have on-site provision of a wide range of relevant investigations and associated specialists, and also shorter waiting times to see new patients with suspected seizures. There were also significant differences between epilepsy clinics and general neurology clinics in the provision of written information and counselling. Epilepsy clinics have definite advantages for patients over general neurology clinics in improving access to investigations and specialists and provision of psychosocial support, but the extent to which these translate into positive health outcomes needs further evaluation. A second survey of directors of public health concerning purchasing arrangements for epilepsy services confirmed that purchasers, as yet, are making little use of the contracting process to influence the quality of epilepsy services offered by providers of care. PMID- 9203247 TI - The biochemical investigation of epilepsy in childhood. AB - Metabolic disorders are a relatively uncommon, but important, cause of childhood seizures. Routinely searching for a metabolic disorder in epilepsy is usually unrewarding. Children and adolescents with epilepsy can be selected for appropriate biochemical investigation by considering the seizure type, age of onset, EEG appearance, family history, clinical findings and the results of brain imaging. A limited number of biochemical screening tests can be used to categorize the more common metabolic disorders associated with seizures. PMID- 9203248 TI - Long-term outcomes of conventional therapy for infantile spasms. AB - Infantile spasms (IS) is an age-specific epilepsy which responds to anticonvulsant therapy but has a generally poor prognosis for normal psychomotor development. The subgroup of infants with a cryptogenic aetiology or whose therapy is initiated promptly is thought to have a more favourable prognosis. We retrospectively reviewed 28 infants with IS treated between 1990 and 1996 with adrenocorticotropic hormone (ACTH), valproic acid (VPA), or both, in order to correlate therapeutic response with long-term outcome. Mean age at onset of treatment was 6.4 months, with 57% of patients started within 1 month of IS appearance. IS was considered cryptogenic in 39%. The majority of infants responded to ACTH or VPA with a reduction in spasms of 75% or more. Total remission of seizures occurred in 52%. Death occurred in eight patients; mean duration of follow-up for survivors was 55 months. All subgroups based on age, aetiology, or treatment had poor outcomes, commonly with residual epilepsy, cerebral palsy or mental retardation. Conventional treatment for IS, even when initially successful in reducing spasms, is inadequate when viewed from a long term developmental perspective, suggesting the need for novel innovative approaches for treating IS. PMID- 9203250 TI - Degree of left hippocampal atrophy correlates with severity of neuropsychological deficits. AB - The relationship between the degree and distribution of hippocampal atrophy measured by volumetric magnetic resonance imaging and severity of memory deficits in 25 patients with temporal lobe epilepsy secondary to mesial temporal sclerosis was assessed. Hippocampal volumes were expressed as a ratio of smaller to larger, normal ratio greater than 0.95. Neuropsychology tests included: subtests of the WAIS-R, Rey Auditory Verbal Learning Task, Rey Figure and the Austin Maze. Degree of left hippocampal atrophy in patients with left temporal lobe epilepsy was associated with severity of verbal memory deficits as measured by RAVLT total recall (P < 0.05), delayed recall (P < 0.001), story recognition (P < 0.001), list recognition (P < 0.001) and final delayed recall (P < 0.001) and recall of the Rey Figure (P < 0.01). There was no association between degree of right hippocampal atrophy and any of the memory tests. Diffuse left hippocampal atrophy was associated with more severe verbal memory deficits than anterior atrophy. We conclude, the association between degree of left hippocampal atrophy and verbal memory provides further evidence of the predominant involvement of the left hippocampus in verbal memory. The finding of a relationship between degree of left hippocampal atrophy and measures of non-verbal function suggests these tests are dependent on verbal memory, or that mesial temporal sclerosis is a bilateral but asymmetrical condition. PMID- 9203249 TI - Basic haematological parameters, serum gamma-glutamyl-transferase activity, and erythrocyte folate and serum vitamin B12 levels during carbamazepine and oxcarbazepine therapy. AB - Basic haematologic parameters and serum gamma-glutamyl-transferase (GGT) activity were evaluated in a five-year prospective follow-up study of 25 patients with newly diagnosed epilepsy starting treatment with carbamazepine. In addition, we evaluated the effects of replacing carbamazepine by oxcarbazepine on these parameters, erythrocyte folate concentrations and serum vitamin B12 levels in 12 male patients with epilepsy. The mean white blood cell count (WBC) and red blood cell count decreased after 2 months carbamazepine therapy, and remained at this lower level during the first 5 years of medication. The mean erythrocyte volume (MCV) and the serum GGT activity increased progressively during carbamazepine treatment. The serum GGT activity decreased after replacing carbamazepine by oxcarbazepine indicating a normalization of the liver P450 enzyme system induction. Concomitantly, the erythrocyte folate concentrations and serum levels of vitamin B12 increased, and the WBC increased and MCV decreased. It is probable that the changes in folate metabolism and serum vitamin B12 concentrations are due to normalization of the liver P450 enzyme system induction after the change of medication. The haematologic changes during carbamazepine medication, and their normalization after replacing carbamazepine by oxcarbazepine are possibly related to changes in folate and vitamin B12 metabolism. PMID- 9203251 TI - Severe myoclonic epilepsy in infancy: evolution of seizures. AB - Changes in seizure type of severe myoclonic epilepsy (SME) in infancy were reviewed retrospectively in 14 patients (11 males and 3 females) who were followed-up to the age of 7 years or more. The observation period ranged from 5 to 16 years with a mean of 10 years. During the follow-up, three or four types of seizures were seen per patient, but the pattern of appearance and disappearance of each seizure type varied considerably among the patients. Tonic-clonic convulsion, either generalized or unilateral, was seen most consistently through the entire course, and it continued to the end of follow-up in 11 patients (79%). On the contrary, myoclonic seizure, complex partial seizure, and atypical absence often disappeared and reappeared repeatedly during the course. In SME, seizure symptoms varied widely among patients in comparison with other neurological symptoms, and the most consistent core seizure type was tonic-clonic convulsions. PMID- 9203252 TI - Multicentre clinical evaluation of vigabatrin (Sabril) in mild to moderate partial epilepsies. French Neurologists Sabril Study Group. AB - Vigabatrin (VGB) has been shown through several studies to be safe and effective as add-on therapy, particularly for the treatment of partial seizures in patients with severe epilepsies followed for years in hospital-based clinics. We now report additional clinical experience with VGB arising from an open trial of add on VGB therapy in patients with relatively few seizures followed by qualified neurologists in private practice (the French Neurologists Sabril Study Group). VGB was administered to 397 patients aged 12-74 years (mean age = 37.5 +/- 13.8 years) who presented with no more than seven partial seizures of any type per month during a 3-month baseline period (mean number of seizures = 3.7 +/- 1.9/month). Simple partial seizures were reported in 121 (30.5%) patients, complex partial seizures in 282 (71.0%) and seizures with secondary generalization were reported in 111 (28.0%). The mean number of associated antiepileptic drugs (AEDs) was 1.9 +/- 0.9 and the mean dose of VGB was 2.21 +/- 0.64 g/day. Following introduction of VGB, 53 (13.4%) became seizure-free and remained so during the whole trial. During the fourth month of treatment, 158 patients (39.8%) had no seizures at all and a further 69 (17.4%) had their seizure frequency reduced by more than 50%. Secondary generalization was controlled during the whole period of treatment in 55 out of 97 patients (56.7%), 17 of which remained free of all types of partial seizures. VGB showed a good tolerability profile; adverse experiences more frequently reported were drowsiness and sleep disturbances. No action was necessary in the great majority of cases; the dose was reduced in 26 (6.5%) and VGB was discontinued in 32 (8%) patients. These data provide additional evidence that VGB can be used safely early on to treat patients with mild to moderate partial epilepsies. Secondary generalization was controlled in the majority of patients. Factors associated with the everyday clinical use of VGB, that resulted from a series of organized meetings with the investigators, are discussed. PMID- 9203253 TI - Obstructive sleep apnoea following rapid weight gain secondary to treatment with vigabatrin (Sabril). AB - A case of a patient with medically intractable epilepsy, who developed obstructive sleep apnoea (OSA), and an increase in seizure frequency, as a consequence of weight gain following treatment with vigabatrin is described. The relationship between exacerbation of epilepsy and sleep disruption secondary to OSA is discussed. Recommendations regarding the choice of anticonvulsants in patients at risk of developing OSA are made. PMID- 9203254 TI - Reversible MRI lesions after seizures. AB - After generalized or partial seizures, transient lesions may appear on magnetic resonance (MR) images. The mechanisms of MR changes might be a defect in cerebral autoregulation and blood-brain permeability. We report a patient with partial and secondary generalized tonic-clonic seizures. After her first seizure which was generalized tonic-clonic in nature, we detected multiple high signal intensities over the frontal cortical area on proton density images which were enhanced with gadolinium on T1-weighted images. The first and repeated EEGs showed no abnormalities or epileptic discharges. We started carbamezapine (600 mg/d) and excluded systemic diseases like vasculitis, infections, aetiological factors causing cerebrovascular diseases. In the follow-up, she was seizure free under antiepileptic therapy and no other neurological deficit. Repeated MR scans after 24 months from her first seizure revealed no pathologic signal intensities. Although the pathophysiology is unknown, recognition of reversible lesions helps diagnostic and therapeutic approaches to abnormal MR findings after seizures. PMID- 9203255 TI - Will vascular surgery survive the millennium? An overview. PMID- 9203256 TI - The changing vascular surgery workforce. AB - An adequate workforce is requisite for efficient and quality surgical care of patients having vascular disease. Data from 1985 and 1992 United States workforce studies provide the basis for certain projections. Vascular surgeons performed 41% of vascular operations in 1985 and 51% in 1992. The basis for this change seems related to the fact that younger general surgeons are less likely to practice vascular surgery than retiring senior general surgeons, whom they are replacing, and vascular surgeons have assumed the care of patients usually treated by the latter group of surgeons. Our aging society compounds this change. Patients older than 65 years will exhibit a 73% increase from 2010 to 2030, with a subsequent greater need for more vascular operations, which are predicted to be 1,020,067 in 2020, compared with 583,000 in 1992. A larger workforce will be required to meet this need. Although innovative technology and changes in health care delivery may cause unpredictable changes in these anticipated workforce needs, the potential exists that there will be insufficient numbers of surgeons available to provide adequate surgical care early in the next century, unless changes occur in the training and practice of general and vascular surgeons. PMID- 9203257 TI - The next generation of vascular surgeons: how should they be trained and credentialed? Who should pay for it? AB - Vascular surgery has matured as a subspecialty of general surgery during the past 25 years. The discipline currently faces a number of challenges including duration and scope of training, financing of graduate medical education, specialty and subspecialty certification, hospital privileging or credentialing, and practice opportunities. This article reviews these issues in terms of present realities and future opportunities. Emphasis is placed on curricular redesign to tailor length and breadth of training to future career goals, be they private practice as a general surgeon with vascular surgery interests, a vascular surgical subspecialist, a rural versus an urban practice, or a subspecialist in an academic setting. A plea is made for regulatory bodies, including the Residency Review Committee for Surgery and the American Board of Surgery, as well as specialty societies, particularly the American College of Surgeons, to maintain a flexible and constructive posture in dealing with proposed educational innovations. With cooperative efforts of both the Association of Program Directors in Surgery and the Association of Program Directors in Vascular Surgery, vascular surgery education and training of both general surgeons and vascular surgeons should be enhanced to fulfill the workforce needs as we enter the next millennium. PMID- 9203258 TI - Leaving the nest: should vascular surgery seek board status separate from general surgery? AB - Current changes that impact on vascular surgery include altered financial support, the introduction of endovascular treatments, and possible reentrance of general surgeons into the vascular field. In addition, vascular surgery in competition with interventional specialties (radiology and cardiology) for patients. These changes and the enhanced competition mandate that vascular surgery and vascular surgeons adapt by becoming competent with endovascular techniques to perform their standard operations better and to provide newer, better treatments that replace standard vascular operations. Vascular surgery must also resolve its conflicts with the American Board of Surgery (ABS) and the Residency Review Committee in Surgery (RRC-S) so that adequate numbers of well trained vascular surgeons will be available to care for vascular disease patients optimally. Whether this can occur by modifying the present system or by having a separate board and residency review committee in vascular surgery will depend on the willingness of the ABS and the RRC-S to recognize that vascular surgery is a separate specialty that is best performed by those with special training and commitment to this field. PMID- 9203259 TI - Endovascular surgery: threat or opportunity? AB - Although the fundamentals of Endovascular Surgery were set down over 30 years ago, only recently has the potential impact of this therapy become readily apparent. Endovascular therapies with new instrumentation for the treatment of arterial aneurysms, occlusions, trauma, and venous thrombotic disorders have progressed dramatically over the past 5 years. Guidewire and catheter techniques for endovascular instrumentation of vessels have also been shown to compliment standard vascular surgical interventions. The need for vascular surgeons to obtain the knowledge and professional skills to perform wire and catheter techniques is essential to convert the professional "threat" of endovascular surgery to a future of "opportunity." PMID- 9203260 TI - A new paradigm: the vascular center. AB - The vascular surgeon, in the absence of effective medical options for the treatment of atherosclerosis, has in the past assumed primary responsibility for the care of patients with peripheral vascular disorders. During the past two decades, management of these patients has become increasingly complex, the direct result of extraordinary proliferation of both our aging population and the expanding technological array of treatment options available. Such patients often require multiple medications with repeated encounters with a variety of specialists resulting in inefficient and not infrequently ineffective care. The concept of a truly integrated multidisciplinary effort to expedite the care of patients with peripheral vascular disease developed in the early 1980s. A vascular center was created at the Brigham and Women's Hospital involving the departments of Surgery, Medicine, and Radiology with their appropriate subspecialty divisions. The combined expertise of dedicated individuals interested in a disease-oriented perspective has resulted in a marked improvement in the efficiency and, it is hopeful, efficacy of management. PMID- 9203261 TI - Cost-effective use of the noninvasive vascular laboratory: potential trends related to increased economic pressures. AB - Because noninvasive studies typically are used in the place of more costly and more invasive technologies, the vascular laboratory remains a key element in the care of vascular patients. We have examined an established model for vascular laboratory cost/efficiency analysis in light of trends toward decreased reimbursement. In this study, we review potential strategies for more cost effective laboratory management, including the impact of part-time technologists, staggered work hours, and equipment allocation. As the need to decrease costs continues, the greatest pressure will be felt in the areas of personnel and equipment costs. Diligence and dedication will be required to prevent these pressures from creating a negative impact on the quality of studies, patient access to care, and technological innovation. PMID- 9203262 TI - How can vascular surgery survive capitated care? AB - The basic differences between fee-for-service and capitated care are characterized and how they are likely to impact the practicing vascular surgeon are explored. Likely practice scenarios under capitated care are given. If the potential problems associated with using primary care "gate keeper" physicians materialize, secondary "carveouts" by vascular specialists may prove feasible. The alternative, particularly in large patient care consortiums, is to educate gatekeepers with efficient management algorythms in exchange for appropriate referrals. In spite of demonstrable flaws and abuses, capitated, managed care is likely to persist in some form, as an effective way to control spiraling health care costs. Therefore, vascular surgeons need to be prepared to compete under such systems. The major keys to success are efficient use of diagnostic tests, applying conservative, cost conscious indications for intervention, and efficient use of hospital resources (judged by lengths of stay in the intensive care unit and hospital and readmissions under the same diagnosis). In addition to developing better practice profiles, in these and standard mortality and morbity parameters, vascular surgeons need to create or join an efficient practice environment, with a minimum number of efficient personnel and teaching and research costs tightly controlled. To negotiate contracts effectively, vascular surgeons must know their costs, of delivering care on a per capita basis in their environment, adjusted for specific (eg, working or retired) patient populations. Outcomes and cost comparisons and other management research must vie for attention with new technology and basic research. PMID- 9203263 TI - Critical pathways and cost-effective practice. AB - With increasing pressure by third-party payers and federal reimbursement systems to reduce health care expenditures, cost-effective means to care for the resource intensive vascular surgical population must be explored. The challenge of meeting these cost-saving priorities while maintaining or improving quality of care can result in conflicting demands on surgeons, particularly in academic practices. The adaptation of an industrial management tool-the critical pathway method-to health care delivery is an attempt to reduce length of stay and improve efficiency. Utilization of vascular nurse practitioners, concentration of vascular patients into a dedicated unit, reduction in angiography through more aggressive use of the vascular laboratory, and optimal use of rehabilitation units and skilled nursing facilities are important adjuncts to reduce length of stay without sacrificing quality of care. This report describes the critical pathway method as implemented in a university teaching hospital, and the integration of other modalities in the care of vascular patients that has reduced length of stay by as much as 40% for some vascular surgical procedures. PMID- 9203264 TI - Resource-Based Relative Value Scale (RBRVS), coding, and Medicare reimbursement. AB - Many changes occurred during 1996 in the Resource-Based Relative Value Scale that determines physician payment for services to Medicare beneficiaries in 1997. These issues include the 5-year review of physician work values, the Correct Coding Initiative, changes in the surgical Conversion Factor, and a new Medicare payment formula. Likewise, several more changes on the horizon in 1997 will dramatically impact the 1998 Medicare Fee Schedule, primarily the upcoming resource-based Practice Cost Relative Value Scale, and possible elimination of the separate surgical Conversion Factors. There are also several new Current Procedural Terminology codes that will receive Medicare payment in 1997 and 1998. This article summarizes these events and issues from a vascular surgical perspective. PMID- 9203265 TI - Morbidity and Irish Catholic descent in Britain: an ethnic and religious minority 150 years on. AB - Ethnic and religious minorities often suffer disadvantages both in socio-economic status and in health. Data from the West of Scotland Twenty-07 study suggest some differences in morbidity between those descended from Irish Catholic migrants of the great emigration from 1840 onwards and others. Catholic religion of at least one parent or at birth is used here as a proxy measure to indicate Irish Catholic descent, on the basis of estimates of sensitivity and specificity in the local area. Higher proportions of "Catholics" are in manual social classes. Differences between "Catholics" and "non-Catholics" in one or more age cohorts are observed for the following aspects of health and physical development: general and physical health (self-assessed health, number of symptoms, accidents), psychological distress (depression, anxiety, number of psychosomatic symptoms), impairments and disabilities (sight, hearing, wearing dentures, disability), and physical measures (height, waist-to-hip ratio, lung function). Furthermore, for all aspects except hearing, wearing dentures and number of psychosomatic symptoms, significant differences remain after accounting for sex and social class. For each measure where a difference is observed, it is those respondents with a Catholic parent or who were born Catholic who experience poorer health or physical development. This suggests that those of Irish Catholic descent are at some disadvantage compared with the rest of the population, with respect to health as well as social class, 150 years after the start of the main migration. PMID- 9203266 TI - HIV-infected women: barriers to AZT use. AB - AZT has become a mainstay drug in efforts to slow disease progression in HIV infected individuals. Further, recent evidence indicates that AZT use by pregnant infected women and their neonates may reduce the risk of vertical transmission. In a study of HIV-infected women's treatment-related behavior, attitudes toward the use of this drug were examined. Data were gathered through unstructured interviewing techniques. The data from the first 71 women accrued revealed that negative attitudes towards its use were widely prevalent. Women viewed the drug as highly toxic, prescribed indiscriminately, inadequately tested in women and minorities, promoted for the wrong reasons and inappropriate while they were feeling well. The findings suggest that removing attitudinal barriers to the use of AZT will be important to both primary and secondary prevention efforts. PMID- 9203267 TI - Negotiating spaces in home environments: older women living with arthritis. AB - Within medical geography there has been a surge of interest in applying critical concepts in social theory to empirical settings, including those for persons with disabilities. The ways through which persons with disabilities negotiate space vary widely according to material and social experiences of being disabled. For older women, chronic illness as a type of disability shapes the way in which they approach their daily lives with respect to both the physical and social aspects of their home environments. In the first half of the paper, conceptually, I take a relational view of space and argue that household, as a narrow reading of domestic space, needs to be replaced by home environment which incorporates more fully age- and ablement-sensitive readings of the spaces constitutive of domestic space. This lays the basis for a contextualized socio-spatial understanding of the ways older women with chronic illness negotiate the spaces in home environments because it accounts for the disadvantaged positionings of access to power and resources as well as the uneven distributions of income based on gender, age, and (dis)ability. It also takes into account the material and social aspects of being disabled. In the second half of the paper, I present case studies of three older women diagnosed with rheumatoid arthritis to illustrate these arguments. PMID- 9203268 TI - Young doctors' health--I. How do working conditions affect attitudes, health and performance? AB - Long hours and other difficult working conditions are thought to affect the health of young doctors, but there has been little evidence to support these assertions. Data are presented from a class cohort of junior doctors in the U.K. showing the relationships between working conditions, health and performance. Long hours appear to have short-term consequences in terms of the doctors feeling unwell and reporting poor performance, as measured by the somatic and social dysfunction scales of the General Health Questionnaire, but there are no demonstrated long-term health consequences. Instead, a number of working conditions, number of emergency admissions, number of deaths on the ward and the number of minor menial tasks contribute to a perception of being overwhelmed, as revealed by factor analysis of the Attitudes to Work questionnaire. This factor correlates significantly with a range of long-term physical and mental health measures as well as measure of work performance. PMID- 9203270 TI - The status of genetic material and genetic information in The Netherlands. AB - The moral status of genetic material and information, and the ethics of controlling and manipulating them, is a topic of hot debate in many European countries, including The Netherlands. That heat is due partly to the complexity of the topic, and partly to researchers' fear that their investigations will be hampered by restrictions on the use of personal data or body material. But there is little doubt that manifold diverging interpretations about the status of the human body, body materials, and personal information in Dutch law, written and unwritten, contribute to the intensity of the debates. This article intends to structure the debate by creating more clarity at the conceptual level. By carefully examining relevant articles of the Constitution and Civil Codes, as well as policy documents and authoritative publications, notably in reference to prominent legal concepts such as property, ownership and privacy, and answer should be provided to the following crucial question: is the status of genetic material and information in any sense special in comparison with other body parts and other kinds of information about a person? This paper first discusses the status of human body materials and personal information in general, and then continues with a more specific discussion about the status of genetic material and information. It concludes that the Dutch legislature had carefully avoided (or not felt the need to employ) the concept of ownership in regulating biomedical research; rather, privacy is found to be the prime regulatory concept. PMID- 9203269 TI - Young doctors' health--II. Health and health behaviour. AB - There is little published information on the health of young doctors, apart from a number of studies which show increased rates of psychiatric symptoms. Nor is there much known of their health behaviour. Anecdotal accounts suggest that doctors' own health care is poor, especially in terms of their willingness to consult other doctors. This paper presents data from a longitudinal study of a class cohort of young doctors first interviewed when they were students. Data show that they suffer from frequent minor physical ailments, with women reporting more ailments than men. Despite this, they took less sick leave. Overall, the doctors took very little time off work. Using the GHQ-28, with a threshold of 5/6, 30% of doctors fell into the "caseness" category for psychiatric symptoms. This is in keeping with findings elsewhere. From the doctors' own reported health behaviour, both in terms of their response to illness over the past year, as well as their predicted response to hypothetical illness, they have developed maladaptive patterns. These include continuing to go to work when unfit, self prescribing, and consulting friends and colleagues rather than going for a formal consultation. This is seen as inappropriate, especially in cases of mental illness. A third of the young doctors are not registered with a local general practitioner and the majority have no clear idea of the role of the Occupational Health Service. The results are discussed in terms of the need to change attitudes to health care and to develop guidelines, staffing and services to enable doctors to take better care of themselves. PMID- 9203271 TI - Ownership of genetic material and information. AB - As a result of the International Human Genome Project genetic information is rapidly multiplying. To avoid some of the problems regarding the availability and use of genetic information, it is sometimes suggested to apply the concept of ownership. This article focuses on the clarification of the status of genetic material and genetic information, obtained as a result of screening and counseling of individual patients. First, some philosophical theories of ownership are examined for a justification of the use of the concept of ownership with regard to the human body. Next, arguments with regard to ownership of the human body are examined. The results of this analysis are applied to genetic material and genetic information. PMID- 9203272 TI - World War I origins of the syphilis epidemic among 20th century black Americans: a biohistorical analysis. AB - Syphilis outbreaks and differentials have been an ongoing issue in modern preventive medicine and public health. Since the early 20th Century, a variety of approaches has been employed to explain demographic and temporal variations in the prevalence of syphilis in the U.S. Public health experts and physicians have tended to rely on case-by-case approaches to explain group-specific patterns. This study, however, shows that population-level disease dynamics cannot be ascertained from these individual-level studies. We offer a biohistorical methodology to study syphilis prevalence differentials in U.S. populations. Using historical health data, this study suggests that the social disruption brought on by World War I was the critical and unique environmental condition which ignited an epidemic of syphilis among black Americans. By establishing this beginning point for the epidemic, this study further shows the persistence of the epidemic for the next 40 years and its decline. This biohistorical methodology could be applied to the analysis of STD epidemics in other populations and regions experiencing mass exposure events. PMID- 9203273 TI - Bore holes and the vanishing of guinea worm disease in Ghana's upper region. AB - Ghana's Upper Region provides an excellent example of the beneficial effects of improved water security provided by hand-pump tube wells. Following a Ghana Canada bilateral development project that installed some 2500 pumps, protection rates against guinea worm disease may be estimated as 88% in the west, and 96% in the east. Survey comparisons between ca 1960 and 1990 show that dracunculiasis declined in 32 of a total of 38 areas. The shadow of guinea worm has been lifted from the land and, in many areas, a true "vanishing" has occurred. The few areas of disease increase are characterized by the lowest population densities, pioneer settlement for cotton farming, and an absence of bore holes. Vagaries of development have inadvertently produced disease transformations or "metamorphoses" from dracunculiasis to elephantiasis (lymphatic filariasis) in one area, and to red water disease (schistosomiasis hematobium) in other areas. Correlative associations between pump densities and guinea worm disease are weakened by the large size of areas for which disease is reported in 1990. One preliminary finding is that geographical distance to the pump is a stronger influence than demographic pressure on pumps, regarding dracunculiasis. Diminishing returns on higher pump densities in many areas support the idea of making fuller, safer use of supplementary non-pump water. Despite crises of fee payment and pump maintenance, the rural bore hole project has struck a mortal blow against guinea worm, and permanently raised the quality of life in the Upper Region. PMID- 9203274 TI - Lay injection practices among migrant farmworkers in the age of AIDS: evolution of a biomedical folk practice. AB - The practice of injecting vitamins and antibiotics by lay people is common among Hispanic migrant farmworkers in the U.S.A. This practice has recent roots in the Latin American cultures from which these farmworkers originate, but it presents a public health concern in its new context because of the high prevalence of HIV infection among this disenfrachised population. Reasons for use of lay injections include cultural beliefs about the superiority of injections over oral forms of medications, perceived irrelevance of a professional diagnostician in prescribing empirical treatment, and a multitude of barriers to access to Western medicine. Although HIV educational materials directed at migrant farmworkers do not address the issue of sharing needles for these types of injections, some farmworkers indicated they had already modified their injection techniques in response to simple directives from physicians in their home country. In contrast to other folk treatment practices that have been resistant to change mediated solely through the provision of information, lay injection is such a new development that considerable experimentation and incorporation of new knowledge are still actively shaping its use. In this process, physicians are seen as legitimate sources of information about the use of Western pharmaceuticals; they should use this role to discourage unsafe injection practices. Efforts to extinguish the practice of lay injection entirely are less likely to meet with success so long as other means of accessing Western medicine are limited. PMID- 9203275 TI - Life stories and shared experience. AB - Illness narratives have become a central issue in medical anthropology. Many researchers have made use of narratives as data in a meaning-centered approach, analysing personal illness accounts as a kind of coping strategy by which human beings ascribe cultural meaning to suffering. Often such narratives are being presented as clinical case stories or as patients' accounts told in interviews to a researcher. But apart from being methodologically created data personal stories also have their own life. They are a way of expressing experience, and as reality manifests itself as experience in us, stories are fundamental to human understanding. In many therapeutic groups personal stories are told as a way of sharing experience in order to solve common problems. This article focuses on the social and processual nature of personal narratives as they are presented in Alcoholics Anonymous (AA) groups. The article is based on a study of AA and Minnesota Model treatment of alcoholism in Denmark from 1990 to 1993. Various genres of personal narratives told at AA meetings are identified and analysed referring to individual as well as social and cultural levels. By focusing on interpersonal relationships and the creation of a shared identity in the groups, the article suggests that the ongoing telling of personal narratives in Alcoholics Anonymous takes place in a continuum between autobiography and myth. Thus, individual and collective experience are merged into the same therapeutic process. PMID- 9203276 TI - Sociocultural factors and the promotion of exclusive breastfeeding in rural Yoruba communities of Osun State, Nigeria. AB - Child survival strategies include prolonged and intensive breastfeeding, together with its early initiation, and breastmilk only for the first six months of life. This paper reports on local knowledge and attitudes of breastfeeding and the sociocultural factors that shape its practice in poor rural Yoruba communities of Southwestern Nigeria. The study has conducted 10 focus group discussions among homogeneous groups of grandmothers, pregnant women, lactating mothers, husbands, and community health workers, and a questionnaire survey of 256 third trimester pregnant women. All women in these communities breastfeed their infants on demand, and for up to two years, because breastmilk is universally accepted as the best food for babies, and breastfeeding spaces births. Prelacteal feedings of water herbal infusions and ritual fluids are the norm, and breastmilk is supplemented, from birth, with water and teas. Exclusive breastfeeding is considered dangerous to the infant: the baby has an obligatory requirement for supplementary water to quench its thirst and promote its normal development, and for herbal teas which serve as food and medicine. Colostrum is discarded because it is dirty, "like pus", and therefore potentially harmful to the infant, although 24% of the survey sample would give it to their babies. Expressed breastmilk is suspect as it can get contaminated, poisoned or bewitched. Complementary foods are introduced as early as two months because of perceived lactation insufficiency. The commonest supplement is a watery maize porridge of low nutrient density. Breastfeeding can also be dangerous, as toxins and contaminants can be passed to the infant through breastmilk. The most serious conflict with the WHO/UNICEF recommendations is the lack of local credibility of exclusive breastfeeding. According to local knowledge, the early introduction of water, herbal teas, and of complementary foods is designed to enhance child survival, while these are supposed to do the exact opposite by the WHO/UNICEF rationale, by exposing the infant to contaminants early, thereby increasing diarrheal morbidity and mortality. Child survival interventions need to address this conflict. PMID- 9203277 TI - Gender differences in correlates of disablement among the elderly in Egypt. AB - The purpose of this paper is to examine gender-specific models to determine whether different combinations of correlates are associated with male and female disablement, using a sample of noninstitutionalized elderly persons in Egypt. Because women and men have different work, family, and household roles, as well as different health risks, it is reasonable to assume that there may be differing correlates for disablement for elderly males and females. The dichotomous dependent variable indicates problems, or the inability, in performing at least one of six activities of daily living (ADL). Of interest is the association of health, economic, and family variables, controlling for age. Logistic regression models are estimated for the total, male, and female samples. The results indicate that for males, having to stop working due to illness and having an unattended medical need are associated with higher odds for disablement as compared with females. Additionally, illiteracy increases the odds for male disablement, yet it has no effect on female disablement. Having experienced an injury in the past year is associated with disability for females, as is spending a lifetime in a rural setting and currently living in a fair to poor residence. Also for the females, increased number of living children significantly increases the odds for functional disability. Separate male and female models were estimated for each of the six ADL items. The trends indicated that the model covariates were more useful in modeling female disabilities in personal care activities, rather than problems with eating and mobility. The health variables were significantly associated with most of the specific ADL problems for the males. PMID- 9203278 TI - Understanding donors' motivations: a study of unrelated bone marrow donors. AB - Medical advances in bone marrow transplantation techniques and immunosuppressive medications have dramatically increased the number of such transplants performed each year, and consequently, the demand for bone marrow from unrelated donors. Although physiological aspects of bone marrow donation have been thoroughly investigated, very few studies have examined psychosocial factors that may impact individuals' donation decisions and outcomes. To examine one particular set of donor psychosocial issues, this study investigated motives for bone marrow donation among 343 unrelated bone marrow donors who donated through the National Marrow Donor Program. Six distinct types of donor motives were identified from open-ended questionnaire responses. Donors most frequently reported motives reflecting some awareness of both the costs (to themselves) and potential benefits (to themselves and the recipient) of donation. A desire to act in accordance with social or religious precepts, expected positive feelings about donating, empathy for the recipient, and the simple desire to help another person were also commonly cited reasons for donating. Among a series of donor background characteristics, donors' gender was the variable most strongly associated with motive type; women were most likely to cite expected positive feelings, empathy, and the desire to help someone. Central study findings indicated that donor motives predicted donors reactions to donation even after the effects of donor background characteristics (including gender) were controlled. Donors who reported exchange motives (weighing costs and benefits) and donors who reported simple (or idealized) helping motives experienced the donation as less positive in terms of higher predonation ambivalence and negative postdonation psychological reactions than did remaining donors. Donors who reported positive feeling and empathy motives had the most positive donation reactions in terms of lower ambivalence, and feeling like better persons postdonation. These finding add substantially to the body of work concerning medical volunteerism generally, and also have important practical implications for the recruitment and education of potential bone marrow donors. PMID- 9203279 TI - Women's health status and use of health services in a rapidly growing peri-urban area of South Africa. AB - Women's health in South Africa and particularly women living in peri-urban areas is being influenced by three major factors. These include the political transition that is occurring in the country, urbanization and the international interest in women's health. Changes in the delivery of health care to the population, and in particular to women are being planned. It is therefore important that data are available for the purpose of planning and evaluation of health services. This paper describes a household survey in which 661 women were interviewed. Socio-demographic patterns of women living in a rapidly urbanizing area were determined and related to health status, use of health services and knowledge of the services. Poverty appeared to be an overriding factor affecting the health of the population. One third of the women were living in unserviced shacks. There was a high rate of unemployment and those who were employed worked in low status jobs and earned very little. Rates of reported acute and chronic illness were lower than described elsewhere in similar household interview surveys. A third of the acute illnesses were due to respiratory disease. Reported rates of diabetes and hypertension were low indicating undiagnosed disease in the area. Being a member of an alliance household-a mixture of family, friends and lodgers-was the main predictor of acute illness. For chronic disease, age and increasing educational status were the main predictors. Knowledge of services apart from those for cervical cancer screening was good. The latter improved with increasing education, urbanization and being a member of an alliance household. As many of the women lived in unserviced areas and had little or no income the provision of infrastructural services and development programs are essential if their health is to be improved. The existing health services need to be developed to provide a comprehensive primary care service with special attention being paid to the health of women. The service should be close to their homes and be affordable. The information gathered in this survey will be used to plan services for women in the area and will act as baseline data for evaluation. PMID- 9203280 TI - Dietary and lifestyle correlates of passive smoking in Hong Kong, Japan, Sweden, and the U.S.A. AB - From epidemiologic studies in several countries, passive smoking has been associated with increased risk for lung cancer, respiratory diseases, and coronary heart disease. Since the relative risks derived from those studies are weak, i.e. relative risk less than two, we investigated whether poorer diets and less healthy lifestyles might act as confounders and be correlated with having a smoking husband on a cross-cultural basis. Characteristics of never-smoked wives with or without smoking husbands were compared between 530 women from Hong Kong, 13,047 from Japan, 87 from Sweden, and 144 from the U.S. In all four sites, wives with smoking husbands generally ate less healthy diets. They had a tendency to eat more fried food but less fruit than wives with nonsmoking husbands. Other healthy traits, e.g. avoiding obesity, dietary cholesterol and alcohol, or taking vitamins and participating in preventive screening were also less prevalent among wives with smoking husbands. These patterns suggest that never-smoked wives with smoking husbands tend to share the same less healthy dietary traits characteristic of smokers, and to have dietary habits associated with increased risk for lung cancer and heart disease in their societies. These results emphasize the need to take into account the potential confounding effects of diet and lifestyle in studies evaluating the health effects of passive smoking, especially since it is known that the current prevalence rates of smoking among men is indirectly associated with social class and education in affluent urban societies. PMID- 9203281 TI - Dynamics of glass-forming di-n-butyl phthalate as studied by NMR. AB - Spin-lattice relaxation times T1 and nuclear Overhauser effect (NOE) enhancement factors for the individual ring carbons in di-n-butyl phthalate (DBF) show that the reorientational correlation function corresponding to the global dynamics in supercooled liquid can be described by a Davidson-Cole distribution. Measurements of proton spin-lattice relaxation times T1 and T1p, as well as 1H NMR spectra at temperatures below the glass transition temperature, Tg, reveal that the same distribution holds also for description of local dynamics in glassy DBF. The activation parameters of the motions detected are derived. PMID- 9203282 TI - 63Cu-31P coupling constants and 63Cu quadrupole couplings from 31P CP/MAS spectra of copper (I)--phosphine complexes with aryldithiocarboxylates or benzoate. AB - Magic-angle spinning 31P NMR spectra of solid [CuS2C-Ph(PPh3)2] 1, [{CuS2C pT}4(PPh3)2] 2, [{CuS2C-Ph}4(PPh3)2] 3. [CuS2C-Ph(dppm)]2 4 and [CuO2C-Ph(dppm)]2 5, (T = tolyl, dppm = bis(diphenylphosphino)methane) were obtained at 109.6 MH2. They consist of distorted quartets from non-equivalent phosphorus atoms and provide approximate values of the indirect spin-spin coupling constant J[63Cu,31P], that are indicative of the covalency of the dithiocarboxylate-copper bonding. The spacing distortions are related to a number of molecular and structural parameters and thereby allow an estimation of the copper quadrupole coupling constant e2qQ/h which, as expected, is smaller for tetra-coordinated (1, 2, 3 and 4) than for tri-coordinated (5) copper sites. The spectrum of 2 has been successfully simulated (including the isotope effects from the less abundant 65Cu isotope) using the full theory for calculation of the spin eigenfunctions of the quadrupolar nucleus. PMID- 9203283 TI - Spin-lattice relaxation in ammonium compounds with a complex molecular dynamics. AB - Expressions are derived for the initial relaxation rate 1/T1 of protons and deuterons of nontunnelling NH4 and ND4 groups reorienting about various symmetry axes in solids. The reorientation rates are modified by a trigonal, tetragonal or monoclinic distortion of the predominantly cubic hindering potential. When the rates differ sufficiently from each other, two T1 minima are observed with a characteristic ratio. Experiments were performed in NH4VO3, (NH4)2S2O8, (NH4)2PtCl4, and their deuterated modifications, which all exhibit two T1 minima. In NH4VO3 and ND4VO3 the relaxation and spectral data agree rather well with the model of trigonal distortion. Also (NH4)2S2O8 has a preferred threefold axis but there, the large tunnel splitting of protons has to be taken into account before an agreement is reached. All the purely reorientational models fail with (NH4)2PtCl4, where, instead, the ammonium groups are proposed to be ordered into domains at low temperatures. The groups inside the domains and boundary regions give rise to the high- and low-temperature T1 minima, respectively. The boundaries are also believed to give rise to the narrow component in the deuteron spectrum at low temperatures. Evidence for a proton tunnelling frequency of 32 MHz is found in (NH4)2PtCl4. PMID- 9203284 TI - Quantitative study of the short range order in B2O3 and B2S3 by MAS and two dimensional triple-quantum MAS 11B NMR. AB - Two-dimensional multiple-quantum magic angle spinning (MQMAS) NMR and MAS NMR of 11B at various magnetic fields, were applied to elucidate the structure of vitreous (glassy) boron trioxide (v-B2O3), vitreous boron trisulfide (v-B2S3) and crystalline boron trisulfide (c-B2S3). These techniques, when combined with computer simulations of the resulting spectra, provide the isotropic chemical shifts and the quadrupole parameters, as well as a quantitative measure of the intensities of various boron resonances. The MAS NMR of v-B2O3 produced overlapping anisotropic lineshapes corresponding to the -1/2<-->1/2 transition in two distinct types of BO3 units with 3(+/-0.08):] intensity ratio. A combination of MAS and the multiple-quantum method resulted in a better resolved, isotropic 11B spectrum of v-B2O3. A remarkable enhancement of resolution of the MQMAS NMR proved instrumental in finding and identifying various impurities present in v B2S3 and c-B2S3. In addition to the resonances from boron in two types of BS3 groups, four other structural units, BOS2, BO2S, BO3 and BS4, were elucidated from the spectra of vitreous and crystalline samples. The effects of various experimental parameters, such as the magnitude of the B0 and B1 fields, on the resolution of the MAS and MQMAS techniques are also shown. PMID- 9203285 TI - Quadrupolar nutation NMR on a compound semiconductor gallium-arsenide. AB - Lattice defects in a compound semiconductor, gallium-arsenide are evaluated by two-dimensional nutation nuclear magnetic resonance. Especially in the case of indium doped gallium-arsenide, analysis of the nutation patterns indicates that the electric field gradient exists in the whole crystal. Asymmetry parameters and quadrupolar coupling constants are determined as approximately 1.0 and 93 kHz, respectively. These results suggest that the whole crystal is under slight strain. Through this work, it is demonstrated that a two-dimensional nutation nuclear magnetic resonance is the useful method to investigate the lattice defects in gallium-arsenide. PMID- 9203286 TI - Dynamic nuclear polarization of nitrogen-15 in benzamide. AB - A 15N dynamic nuclear polarization (DNP) experiment is reported in which a 15N DNP enhancement factor of approximately 2.6 x 10(2) is obtained on free radical doped samples of 99% 15N labeled benzamide. The free radicals BDPA (1:1 complex of alpha, gamma-bisdiphenylene-beta-phenylallyl with benzene) and DPPH (2,2-Di(4 tert-octylphenyl)-1-picrylhydrazyl) are used as dopants and the spin relaxation effects of adding these dopants are studied by means of changes in proton and nitrogen T1 values of the samples. The combination in solids of a very low natural abundance, 0.37%, a small gyromagnetic ratio, and a long spin-lattice relaxation time for 15N nuclei create severe sensitivity problems that, in large part, are ameliorated by the signal enhancement observed in the 15N DNP experiment on samples containing free electrons. PMID- 9203287 TI - Development and preliminary validation of a microtiter plate-based receptor binding assay for paralytic shellfish poisoning toxins. AB - More than 20 countries have either established or proposed regulatory limits for one or more of the paralytic shellfish poisoning (PSP) toxins as they occur in seafood products. PSP toxin levels are generally estimated using the standard AOAC mouse bioassay, yet because of various limitations of this method [e.g. high variability (+/-20%), low sensitivity, limited sample throughput and use of live animals], there remains a need for alternative testing protocols. A sensitive and selective, high capacity assay was developed for the PSP toxins which exploits the highly specific interaction of these toxins with their biological receptor (i.e. voltage-dependent sodium channel) and is thus based on functional activity. This receptor binding assay provides a radioactive endpoint, and is performed in a microtiter filter plate format with results determined by standard liquid scintillation counting within 24 hr. The Ki for the assay is 3.66 +/- 0.86 nM saxitoxin, with a limit of detection of c. 5 ng saxitoxin/ml in a sample extract. Good quantitative agreement of the assay with both mouse bioassay and high performance liquid chromatographic analysis of crude extracts of contaminated shellfish, as well as PSP toxin-producing algae, was observed. Our findings indicate that the receptor binding assay has a strong predictive value for toxicity determined by mouse bioassay, and that this approach warrants consideration as a rapid, reliable and cost-effective alternative to live animal testing for detection and estimation of PSP-related toxicity in seafood and toxic algae. PMID- 9203288 TI - Biologically active polypeptides in the venom of the jellyfish Rhopilema nomadica. AB - A tropical jellyfish, Rhopilema nomadica (Scyphozoa, Rhizostomeae) has recently invaded the eastern Mediterranean. Its painful stings have been the bane of bathers and fishermen from Egypt to Turkey. This paper reports on the presence of haemolytic activity and alpha-chymotrypsin-like serine protease activity in the venom of the R. nomadica nematocysts. In addition, the presence of phospholipase A2 activity, which has been described previously, is confirmed. Some properties of these activities are defined. PMID- 9203289 TI - Distinct bothrojaracin isoforms produced by individual jararaca (Bothrops jararaca) snakes. AB - Bothrojaracin (apparent mol. wt 27,000) is a potent inhibitor of thrombin previously isolated from the venom of Bothrops jararaca. Several molecular variants (isoforms) have been identified in a pool of venom collected from a large number of animals. In order to determine whether an individual snake produces a single type of bothrojaracin or multiple isoforms, we analyzed the bothrojaracin content of venoms collected individually from six adult B. jararaca snakes. Bothrojaracin was found in all venoms, but its activity was especially high in three of them. After purification on an alpha-thrombin affinity column, followed by gel filtration on Superose 12 HR, proteins from these three venoms migrated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis as single bands of 27,000 and their amino-terminal sequences (residues 1-28) revealed extensive homology with bothrojaracin. In contrast, the material purified from the three venoms with low bothrojaracin activity consisted of bothrojaracin together with inactive proteins. Differences in the sequences obtained for bothrojaracin isolated from individual venoms indicated the existence of more than one isoform in the venom of an individual snake. PMID- 9203290 TI - Activities of cobra venom cytotoxins toward heart and leukemic T-cells depend on localized amino acid differences. AB - Several studies have suggested that along the concave surface of cobra venom cytotoxins, a hydrophobic region flanked by positively charged amino acid side chains, as well as by tyrosine and/or serine/threonine, allows these toxins to depolarize muscle or cause cytolysis. Comparison of biological activities among structurally homologous toxins, however, has revealed significant functional diversity. The objective of the present study was to examine several toxins purified from different cobra venoms with regard to their ability to bind to and kill human T-lymphocytes and rat heart cell myoblasts. The activities observed were then correlated with differences in amino acid residues which occur in restricted regions of the toxins. The absence of an aromatic residue at position 11 (Loop 1) resulted in a lower cytolytic response at every concentration tested. A simple inversion of two residues in the amino acid sequence of toxin Loop 3 selectively impaired heart cell binding and cytolysis, but had no effect on T cells. Loss of a positively charged residue in the tip of Loop 2 minimally affected binding but significantly reduced cytolysis. Replacement of valine at positions 27 and 32, along with the introduction of a negative charge at the tip of Loop 2, interfered with binding to either cell type and caused a reduction in cytolysis. The results of this study suggest that no one loop or region is solely responsible for the toxin's biological activity. However, because the binding and cytolytic sites within these toxins are distinct, it may become possible to develop toxin derivatives in which only selected activities are enhanced. PMID- 9203291 TI - Venom yields from several species of colubrid snakes and differential effects of ketamine. AB - The composition of rear-fanged colubrid snake venoms is largely unknown due primarily to the difficulty involved in venom collection. Several different methods have been used to maximize the yield of Duvernoy's secretions. The method proposed by Rosenberg in 1992, which includes the use of ketamine hydrochloride anesthetic and pilocarpine to induce Duvernoy's glands secretion, was used in the present study to collect venom from eight species of colubrids. Protein concentrations, using a dye-binding microassay technique, were determined for the venoms collected. Average protein concentrations ranged from 49.8 to 96.4%. Most yields (dry weight/snake) obtained from specimens in this study were significantly greater than yields previously reported. There was a wide range of effects that occurred due to the ketamine injections; however, all snakes recovered from the effects of the ketamine hydrochloride/pilocarpine with no apparent ill effects. Recommended doses of ketamine hydrochloride have thus been adjusted, depending on previous reactions to the drug. The use of ketamine/pilocarpine in the collection of Duvernoy's secretion has proven to be highly effective in increasing yields. Some caution should be observed when administering ketamine to various species of colubrids, as effects do not necessarily scale to body mass. PMID- 9203293 TI - Characterization of the two myotoxin a isomers from the prairie rattlesnake (Crotalus viridis viridis) by capillary zone electrophoresis and fluorescence quenching studies. AB - The two myotoxin a isomers from the venom of the prairie rattlesnake Crotalus viridis viridis have different isoelectric points, as determined by capillary zone electrophoresis. The pI values are 10.50 and 10.57, respectively, and both are higher than the previously reported pI value for myotoxin a. The difference in the isoelectric points between the two isomers is attributed to altered surface charge as a result of the conformational change in myotoxin a. Both isomers exist in crude venom, discounting the possibility that they are artifacts formed during the purification process. Fluorescence quenching of myotoxin a reveals heterogeneity of the tryptophans, possibly due to different environments. The fraction of the total tryptophan fluorescence quenched by iodide is 81% and is attributed to solvent-accessible tryptophan residues at the protein surface. The 19% non-quenchable tryptophans probably represent residues that are shielded from the solvent exposure. The ratio of buried to exposed tryptophans is similar to the ratio of isomers seen by capillary zone electrophoresis and reverse-phase high-performance liquid chromatography (c. 1 : 4). PMID- 9203292 TI - Effects of repeated injections of palytoxin on lymphoid tissues in mice. AB - Sublethal doses of palytoxin were i.p. injected repeatedly to mice, and the effects on lymphoid tissues were examined. The weight and morphology of the thymus were influenced during exposure but had generally recovered after 1 month of withdrawal. The ratio of lymphocytes to total leukocytes in blood was decreased during the injection term, and did not recover to a normal level even after 1 month of withdrawal. The component of B-cells in the lymphocytes was clarified as being responsible for the small number of lymphocytes in the recovery process. PMID- 9203294 TI - Time factor in the detection of circulating whole venom and crotoxin and efficacy of antivenom therapy in patients envenomed by Crotalus durissus. AB - Thirty-seven patients envenomed by Crotalus durissus were classified into three groups according to the interval between the bite and hospital admission (delta T): group 1 (n = 14, delta T < 4 hr), group 2 (n = 14, delta T > 4 hr < 8 hr) and group 3 (n = 9, delta T > 8 hr). Venous blood from these patients was sampled for biochemical and hematological analysis and for whole venom, crotoxin and antivenom enzyme-linked immunosorbent assays before antivenom treatment (T0) and at 1 hr (T1), 6 hr (T6), 12 hr (T12) and 24 hr (T24) after the start of antivenom therapy. The patients were treated with 100-200 ml (10-20 ampules) of C. durissus antivenom. Whole venom and crotoxin were detected in 13 (92.8%) and 11 (78.6%) of 14 group 1 patients, respectively, in 11 (78.6%) and six (42.9%) of 14 group 2 patients, respectively, and in two (22.2%) and one (11.1%) of nine group 3 patients, respectively, before antivenom treatment. Data from this study show that whole venom and crotoxin were not detected in most of patients when the time elapsed between the bite and hospital admission was greater than 8 hr, and crotoxin was not detected in most of the patients who were admitted to the hospital at times ranging from 4 to 8 hr after the snakebite. Plasma whole venom, crotoxin and antivenom levels measured over time in these patients show the efficacy of antivenom treatment, since circulating venom and crotoxin were no longer detected 1 hr after antivenom therapy and high antivenom titers persisted for at least 24 hr after serotherapy. PMID- 9203295 TI - Absence of tetrodotoxins in a captive-raised riparian frog, Atelopus varius. AB - Bufonid frogs of the genus Atelopus contain two classes of skin toxins, namely the steroidal bufadienolides and the water-soluble tetrodotoxins. Frogs of the Panamanian species Atelopus varius have now been raised in captivity and levels in skin extracts of bufadienolides and of tetrodotoxin-like compounds assessed, using inhibition of [3H]ouabain binding and inhibition of [3H]saxitoxin binding, respectively. Levels of ouabain equivalents, corresponding to bufadienolides, were comparable to those found in wild-caught frogs from the same population in Panama, while tetrodotoxin-like activity was undetectable. The results strongly implicate environmental factors, perhaps symbiotic microorganisms, in the genesis of tetrodotoxins in the skin of frogs of the genus Atelopus, while indicating that the frog itself produces the skin bufadienolides. PMID- 9203297 TI - Purification and sequence determination of a new neutral mammalian neurotoxin from the scorpion Buthus martensii Karsch. AB - A new neutral mammalian neurotoxin, designated BmK M4, with an isoelectric point (pI) of 7.6 and a relatively low toxicity (LD50 = 4.0 +/- 0.25 microgram/g mice, i.v.) was purified from the venom of scorpion Buthus martensii Karsch (BmK). The complete amino acid sequence of the toxin composed of 64 amino acid residues was determined by automated Edman degradation of the N-terminal part of the reduced and S-carboxamidomethylated protein (up to 30 amino acid residues) and its peptide fragments degraded by lysylendopeptidase or Staphylococcus aureus Va protease. The calculated mol. wt based on the amino acid composition was 7001. By comparison with the sequences of other basic BmK mammalian neurotoxins, it was concluded that the weaker toxicity and lower pI value of BmK M4 might be the result of mutations H10E, R18G and K28D. The sequence comparison of BmK M4 with an acidic toxin, BmK M8, showed that the weakest toxicity and acidic property of BmK M8 may be the consequence of mutations K8D, D53A, V55E and V59E. The substitution of 21 Gly in BmK M4 for Tyrin other BmK toxins may also be of importance. In their tertiary structures, these mutated charged residues are mainly distributed in the surface (face B) that is roughly opposite to the "conserved hydrophobic surface" (face A) proposed by Fontecilla-Camps et al. in 1982. Therefore the toxin-receptor interaction may take a multiposition mode. PMID- 9203296 TI - Hypertension and identification of toxin in human urine and serum following a cluster of mussel-associated paralytic shellfish poisoning outbreaks. AB - Following four outbreaks of paralytic shellfish poisoning on Kodiak Island, Alaska, during 1994, medical records of ill persons were reviewed and interviews were conducted. Urine and serum specimens were analyzed at three independent laboratories using four different saxitoxin binding assays. High-performance liquid chromatography was used to determine the presence of specific toxin congeners. Among 11 ill persons, three required mechanical ventilation and one died. Mean peak systolic and diastolic blood pressure measurements were 172 (range 128-247) and 102 (range 78-165) mmHg, respectively, and blood pressure measurements corresponded with ingested toxin dose. All four different laboratory methodologies detected toxin in serum at 2.8-47 nM during acute illness and toxin in urine at 65-372 nM after acute symptom resolution. The composition of specific paralytic shellfish poisons differed between mussels and human biological specimens, suggesting that human metabolism of toxins had occurred. The results of this study indicate that saxitoxin analogues may cause severe hypertension. In addition, we demonstrate that saxitoxins can be detected in human biological specimens, that nanomolar serum toxin levels may cause serious illness and that human metabolism of toxin may occur. Clearance of paralytic shellfish poisons from serum was evident within 24 hr and urine was identified as a major route of toxin excretion in humans. PMID- 9203298 TI - Identification of Caribbean ciguatoxins as the cause of an outbreak of fish poisoning among U.S. soldiers in Haiti. AB - On 24 February 1995, six U.S. soldiers serving with the Multinational Force in Haiti became ill after eating a locally caught fish identified as the greater amberjack Seriola dumerili. The victims presented with nausea, vomiting, watery diarrhea and abdominal cramps 5-8 hr after consumption. Also present in some victims were numbness in the extremities or perioral region, bradycardia and scalp paresthesia. Patients were treated with i.v. hydration therapy and antiemetics. All recovered without sequelae over the course of 1-3 months. A portion of the cooked fish was obtained for analysis. A semipurified lipid extract was prepared according to standard methods and analyzed for the presence of Na+ channel site 5 binding activity using a brevetoxin receptor binding assay. By this assay, the fish sample contained the equivalent of approximately 20 ng Caribbean ciguatoxin/g flesh. The presence of the major Caribbean ciguatoxin (C CTX-1) was confirmed by liquid chromatography-mass spectrometry. Using the receptor binding assay to monitor activity in TSK and PRP-1 column fractions, two minor toxins were detected in addition to C-CTX-1. One of these minor toxins was more polar, and the other less polar, than C-CTX-1. These data provide firm evidence that a family of C-CTX-1 is responsible for ciguatera in the Caribbean. PMID- 9203299 TI - Role of tumor necrosis factor and nitric oxide in the cytotoxic effects of Clostridium difficile toxin A and toxin B on macrophages. AB - Clostridium difficile, the bacterium involved in antibiotic-associated colitis, produces two exotoxins, toxin A (TxA) and toxin B (TxB). Although these toxins are well recognized as being cytotoxic to several mammalian cell types, the mechanisms involved are not fully understood. The aim of the present investigation was to examine the cytotoxicity of TxA and TxB to peritoneal macrophages in culture and to investigate whether tumor necrosis factor-alpha (TNF-alpha) and nitric oxide (NO) are involved in the process. As a control, the effect of E. coli LPS was also investigated. TxA, TxB and LPS were dose dependently cytotoxic to macrophage monolayers, with TxB being the most potent. All of the toxins stimulated the release of TNF-alpha from macrophages. TxB was again the most active in inducing this response. The TNF-alpha released appears to be involved in the action of LPS and TxA, but not of TxB, since a mAb against TNF-alpha inhibited the cytotoxicity of the former two but had no effect on the latter. NO is not involved in the effects of TxA and TxB since these toxins did not induce the production of this mediator in macrophages, even in the presence of IFN-gamma. In addition, L-imino-ethyl-L-ornithine (L-NIO), a NO synthase inhibitor, did not modify the macrophage death caused by TxA or TxB. Although LPS was able to induce the production of high amounts of NO, NO did not mediate the LPS cytotoxicity since L-NIO did not influence the degree of macrophage death caused by LPS. TxA and TxB therefore appear to exert cytotoxic effects on cultured macrophages by different mechanisms. TNF-alpha is involved in TxA and LPS-mediated cytotoxicity but not in the toxicity caused by TxB. NO is not involved in the killing action of any of these toxins. PMID- 9203301 TI - Clinical description of Parabuthus transvaalicus scorpionism in Zimbabwe. AB - An epidemiological and clinical study of Parabuthus transvaalicus scorpionism was conducted in Zimbabwe. Ten per cent of stings resulted in severe scorpionism. The clinical features of 17 patients with severe envenomation were primarily neuromuscular, with significant parasympathetic nervous system and cardiac involvement. The clinical course was prolonged compared to other scorpion syndromes, and significant therapeutic benefit was demonstrated in terms of hospital stay in response to species specific antivenom. The case fatality rate was 0.3%, with deaths in children below 10 years and adults above 50 years. The mortality rate in the district was 2.8 per 100,000 per year. This syndrome from a buthid scorpion resembles in many respects buthid scorpionism described elsewhere in the world, but shows important differences, notably cardiac involvement in the absence of clinical evidence of circulating catecholamines. The relevance of these findings to buthid scorpionism generally are presented as a hypothesis, in which it is postulated that the cardiac effects of the toxins are direct and primary, and autonomic effects secondary but synergistic, determining the ultimate clinical picture. PMID- 9203300 TI - Macrocyclic trichothecenes in Baccharis coridifolia plants and endophytes and Baccharis artemisioides plants. AB - Toxic disease in livestock caused by the shrubs Baccharis coridifolia and Baccharis artemisioides is very common in Argentina. The toxicity of Argentinian and Brazilian B. coridifolia plants and of Argentinian B. artemisioides was investigated. The toxicogenic capacity of 15 endothyte isolates of Ceratopicnidium baccharidicola from B. coridifolia was determined. Roridins and verrucarins were analyzed by thin-layer chromatography using a modified Jarvis method. One-hundred per cent of Argentinian B. coridifolia plants were positive for roridins (RA and RE) and verrucarins (VA and VJ), 16.2% for RD and 2.7% for RH. All of the Brazilian B. coridifolia plants were positive only for roridins. In B. artemisioides plants, RA, RE and RD were present in higher concentrations than VA and VJ, and all of them were more concentrated than in B. coridifolia. One-third of the endophyte isolates were toxicogenic for the same roridins and verrucarins, but in very low concentrations. This is the first report of macrocyclic trichothecenes in B. artemisioides, and a new report of B. coridifolia macrocyclic trichothecenes in Argentina. PMID- 9203302 TI - Lack of effect of endogenous corticosteroids on the acute inflammatory reaction (edema) induced by Bothrops jararaca venom (BjV) in rats. AB - Intraplantar injection of 5 or 10 micrograms of BjV caused local edema in rats that was not affected by ablation of adrenal glands. In addition, no changes in plasma corticosterone levels were observed. Simultaneous injections of the venom into both hindpaws of normal animals, or injections made at varying intervals, resulted in local inflammatory reactions of comparable time-course development and analogous magnitude. These data might be related to an inability of the venom to evoke secretion of corticosteroids. PMID- 9203304 TI - Bibliography of toxinology. PMID- 9203303 TI - Studies on the effect of peroxisomicine on catalase activity in albino mice. AB - Peroxisomicine is a toxic compound isolated from plants of the genus Karwinskia (Rhamnaceae). This toxin produces irreversible and selective damage to the peroxisomes of yeast cells in vivo. Peroxisomicine also inhibits catalase activity in vitro, when using purified enzyme. This paper reports on the effect of peroxisomicine on liver catalase in tissue fragments, in situ, as well as in mice intoxicated with peroxisomicine, in vivo. The catalase activity was determined by biochemical and histochemical methods. In contrast with the reported findings in vitro, the results demonstrate that there is no inhibition of the activity of tissue catalase, and suggest that catalase in situ and in vivo is protected against the inhibitory effect of peroxisomicine by an unknown factor. PMID- 9203305 TI - Field evaluation of an indirect ELISA for detection of brucellosis in lowland Bolivia. AB - Bovine brucellosis exists endemically at an estimated prevalence of 10% in the developing dairy industry of Santa Cruz in tropical Bolivia. This paper describes field testing of an FAO/IAEA indirect ELISA for brucellosis, as a possible replacement confirmatory test for the complement fixation test (CFT). The ELISA and CFT were compared on sera from 3 cattle populations: a non-vaccinated negative population, an S19-vaccinated negative population, and a brucellosis positive population of unknown vaccination status. The CFT and ELISA showed excellent specificities of 100% and 98% respectively against the negative non vaccinated group. The CFT maintained a specificity of 98% against the S19 vaccinated negative group, but ELISA specificity fell to 83% using a cut-off of 20% of positive control, and 94% using a cut-off of 40% of positive control. Against sera from the positive population, the ELISA gave many more positive reactions than the CFT, probably a combination of both higher sensitivity and lower specificity. It is concluded that as Santa Cruz is entering a phase of brucellosis control rather than eradication, the extra sensitivity of the ELISA is not valuable enough to risk a higher level of false positive reactions, especially as S19 vaccination is being increasingly used. PMID- 9203307 TI - Immunisation of smallholder dairy cattle against anaplasmosis and babesiosis in Malawi. AB - A field study was conducted in the Southern Region of Malawi to evaluate the possible benefits of immunisation of improved dairy cattle against Anaplasma marginale, Babesia bigemina and Babesia bovis. Friesian crossbred heifers were immunised when they were being reared on Government farms. They were then issued to smallholder farmers, together with unvaccinated controls, where many of them were exposed to heavy tick infestation. Vaccination was shown to provide a significant degree of protection against babesiosis on the smallholder farms; 15/32 unvaccinated controls developed clinical disease as compared to only 3/28 vaccinates. PMID- 9203306 TI - Use of dot-immunobinding assay for visual detection of rinderpest antibodies in vaccinated cattle. AB - A dot-immunobinding (DIB) assay was used to detect rinderpest antibodies in cattle vaccinated with Kabete 'O' strain vaccine, using purified rinderpest virus. Of 120 serum samples from vaccinated and non-vaccinated animals, rinderpest antibodies were detected in 80%, 88.4% and 91.6% of samples at 2, 3 and 4 weeks postvaccination respectively. All the serum samples from non vaccinated animals were negative. The DIB results had a good correlation with those of the micro neutralisation test. The technique is simple, easy to perform and suitable for routine use in detecting rinderpest antibodies following vaccination. PMID- 9203308 TI - Prevalence of antibodies against respiratory viruses (parainfluenza virus type 3, respiratory syncytial virus, reovirus and adenovirus) in relation to productivity in Syrian Awassi sheep. AB - Awassi sheep sera from all the provinces of the Syrian Arab Republic were tested for RSV, P13, REO and Adeno viruses (IIF and AGID tests). RSV was the most prevalent infection with 63.6% of samples seropositive, followed by REO, P13 and Adenovirus with seroprevalences of 27.3%, 24% and 8.1% respectively. Animals were more frequently infected by RSV alone. Mixed infections were also identified but the occurrence was not high. The RSV and REO virus infections occurred more frequently when transhumant flocks travelled for long distances (P < 0.001). The prevalence of RSV, P13, REO and Adenovirus infections was higher in animals sheltered in poor conditions compared to those in good shelter or with no shelter (P < 0.001; P < 0.001; n.s.; P < 0.05, respectively). RSV and P13 infections were related to an adult mortality rate of 10 to 20% (P < 0.001); REO virus was also proportionally related to mortality from low to high rates (P < 0.05). Concerning mortality of lambs, only Adenovirus infection was related to losses (> 20%) (P < 0.001). REO virus was related to low milk yield (P < 0.05). PMID- 9203309 TI - Immunisation of cattle in Zimbabwe using Theileria parva (Boleni) without concurrent tetracycline therapy. AB - Five hundred and ten cattle were immunised using the Theileria parva (Boleni) stock without concurrent chemotherapy with tetracycline on 2 farms in Zimbabwe, both of which had a history of theileriosis. The stabilate had been titrated in Friesian calves to determine a 50% protective dose (PD50) and 2 or 3 (PD50s) were used to immunise the cattle. None of the cattle showed a clinical reaction following the immunisation procedure. However, the cattle were shown to have responded immunologically on testing for antibodies to a T. parva antigen in an indirect fluorescent antibody test. The immunised cattle were then exposed to a natural field challenge causing severe theileriosis in control cattle. Immunisation against theileriosis without the need for concurrent chemotherapy is much less expensive than the infection and treatment method (US $2.72) compared to US $10.23 in the first year) and would be much more attractive to commercial and traditional farmers. PMID- 9203310 TI - Comparative efficacy of three anthelmintics against mixed gastrointestinal nematode infections in camels. PMID- 9203311 TI - Dermatophilosis and abscessation of lymph nodes in a group of tick-infested horses in Uganda. PMID- 9203312 TI - Coagglutination test for rapid detection of infectious bursal disease virus antigen. PMID- 9203313 TI - Genetic and environmental trends in a large commercial Jersey herd in the Central Rift Valley, Kenya. AB - Records from a large commercial Jersey herd covering the period 1980 to 1993 were used to estimate genetic and environmental trends in production traits, fertility traits and age at first calving. The farm is located in the upper midland agroecological zone of the Central Rift Valley in Kenya. The estimated annual genetic trends were 0.8 kg (milk yield), 0.003% (butterfat per cent), 0.09 kg (butter fat yield), 0 d (calving interval), -0.001 (number of services per conception), 0.22% (conception rate) and -0.5 d (age at first calving). All the traits studied showed an environmental trend in the desired direction: 14.6 kg (milk yield), 0.029% (butterfat per cent), 1.31 kg (butterfat yield), -2.52 d (calving interval), -0.62 (number of services per conception), 0.89% (conception rate), and -17 d (age at first calving). Despite great genetic variation in the production traits, the genetic trend in all traits was very small reflecting ineffective selection, whereas the results for the environmental trends indicate a substantial improvement in the management. PMID- 9203314 TI - Effect of age and sex of egg strain chickens and of two stocking densities of broilers on voluntary water intake and performance in a sub-humid environment. AB - Voluntary water intake and performance of egg-strain chickens of different age groups and sex were compared. Cockerels grew faster and consumed 41% more feed and 43% more water than pullets of the same strain and age. Cockerels of nineteen weeks consumed significantly more feed (P < 0.05) than layers of 30 weeks of the same strain, but there was no significant difference in their daily voluntary water intake. The physiological activities associated with egg laying in chickens, apparently require more water to feed ratio than for the active growth of cockerels. In a second experiment, the effect of 2 stocking densities on voluntary water intake and performance were compared. Birds on higher stocking density (0.09 m2/bird) performed similarly to those on low stocking density (0.18 m2/bird), but the higher stocking density birds recorded slightly higher mortality. Voluntary water consumption was not significantly affected (P > 0.05), although birds on higher stocking density recorded slightly higher average daily voluntary water consumption. The higher stocking density birds recorded slightly higher per cent mortality during the 8 week period, but higher per cent mortality was recorded with the low stocking density birds during the 0 to 4 week brooding period. PMID- 9203315 TI - Estimation of liveweight from chest girth in pure and crossbred west African goats. AB - Linear and geometric regression equations were used to estimate liveweight from chest girth in 78 West African Dwarf and 73 Sahel x West African Dwarf goats. The coefficients of determination (R2) ranged from 0.87 to 0.92 and 0.97 to 0.99 for linear and geometric equations respectively. There were no significant breed or sex differences in the R2 values of the equations. The geometric equations estimated liveweight with a high degree of reliability regardless of girth size whilst the linear equations yielded very low and frequently negative liveweight estimates where girth measurements were below 30 centimetres. PMID- 9203316 TI - Ocular signs in four dogs with hypertension. AB - Four dogs suddenly developed visual problems associated with retinal detachment and haemorrhage. They all had a high arterial blood pressure measured with a Doppler ultrasonic blood pressure monitor. Two of the dogs showed evidence of cardiac hypertrophy and one had hypercholesterolaemia; in three of them there was no conclusive evidence of underlying systemic disease. The condition was treated and assessed over a period of 12 months. In the absence of any findings suggesting the presence of underlying systemic disease, it is possible that the ocular changes were the result of primary hypertension, although not every possible cause of secondary hypertension could be excluded. PMID- 9203317 TI - Percutaneous ultrasound-guided abomasocentesis in cows. AB - The goal of this study was to determine the optimal location for ultrasound guided centesis of the bovine abomasum and to assess the safety of the procedure. In the first part of this study, the technique was applied to 50 clinically healthy cows which were slaughtered within two hours of the procedure. The abomasum and peritoneum were then examined for lesions. In all but one cow, the location for abomasocentesis was 10 to 27 cm caudal to the xiphoid and on the ventral midline or up to 10 cm to the right of it. No peritoneal lesions were observed in any of the cows. In all cases, the site of centesis was visible as a localised haemorrhage on the serosal surface of the abomasum. In 41 of the cows, a haematoma was visible on the mucosal surface of the abomasum. In the second part of the study, 10 cows were monitored clinically for 10 days after abomasocentesis, to assess the safety of the procedure. The appetite, general behaviour, attitude and rectal temperature of the cows remained normal. The haematocrit, total and differential leucocyte counts, and the concentrations of total solids and fibrinogen were determined daily and remained within their normal ranges. At slaughter minimal changes, such as localised reddening and adhesions between the site of the puncture in the abomasum and the abdominal wall, were visible in three of the cows. PMID- 9203318 TI - Transmission of Brucella melitensis from sheep to lambs. AB - Forty-one pregnant sheep showing positive immune responses to Brucella melitensis in serological or allergic tests were selected from naturally infected flocks and kept in an isolated pen for lambing. The resulting 62 lambs were maintained in the same pen with their dams during lactation. When the lambs were weaned, the dams were slaughtered for bacteriological study and the lambs were reared in a clean pen. Fourteen ewes excreted B melitensis during lactation and 17 were found to be infected postmortem, B melitensis was not isolated from seven lambs (three born to infected dams) which died after birth or from eight seronegative lambs (four born to infected dams) which were slaughtered between two and seven months after weaning. However, one permanently seropositive lamb born to a culture negative dam was found to be infected when necropsied five months after weaning. The remaining 46 lambs were reared until adulthood and slaughtered at intervals for bacteriological study. Four ewe lambs (two born to culture-negative dams) were found to be infected postmortem, but were negative in immunological tests for B melitensis. PMID- 9203319 TI - Enzyme-linked immunoelectrotransfer blot assay for diagnosis of hydatidosis (Echinococcus granulosus) in sheep. PMID- 9203320 TI - Commercial polyester fabric repair of abdominal hernias and defects. PMID- 9203321 TI - Isolation of Streptococcus equi subspecies equi (strangles agent) from an Ethiopian camel. PMID- 9203322 TI - Caseous lymphadenitis: an increasing cause for concern. PMID- 9203323 TI - Caseous lymphadenitis: an increasing cause for concern. PMID- 9203324 TI - Avermectin toxicity in the dog. PMID- 9203325 TI - Acute tailhead trauma in Friesian Holstein cows. PMID- 9203326 TI - 'Gut-tie' in steers. PMID- 9203327 TI - Ovine ostertagiosis in the spring. PMID- 9203328 TI - Lead poisoning in swans. PMID- 9203330 TI - Characterization of the role of side-chain interactions in the binding of ligands to apo trp repressor: pH dependence studies. AB - The pH dependence of the association of apo trp repressor with the series of ligands, tryptophan, tryptamine, indole propionic acid (IPA), and trans-beta indole acrylic acid (IAA), has been studied using fluorescence titrations and isothermal titration microcalorimetry (ITC). The purpose of such a comparison of ligands and the pH dependency studies is to reveal the role played by the side chain functional groups in the energetics of the binding of the ligands to the protein. We find that, whereas the binding of tryptamine and IPA have essentially no pH dependence between pH 6 and 10, the binding of tryptophan and IAA depends on pH. For IAA, the affinity drops between pH 6 and 10, consistent with a shift in pKa of some group on the protein from a value of pKa 7.4 to 7.9 upon binding of this ligand. The affinity of IAA also drops below pH 5, but shows saturable binding at pH 2-3, where the protein has previously been found to exist as a partially folded monomeric state. For tryptophan, the pH dependence data indicate that the equilibrium is complicated. We present a model to describe the data in which the alpha-ammonium group of tryptophan has its pKa shifted upward upon binding (i.e. preferential binding of the protonated form of this functional group) and in which the pKa of an unknown group on the protein also has its pKa increased. PMID- 9203329 TI - Light quenching of pyridine2 fluorescence with time-delayed pulses. AB - We describe the effects of time-delayed long-wavelength pulses on the intensity and anisotropy decays of pyridine2. The sample was exposed to a continuous train of 360 nm excitation pulses and time-delayed 720 nm pulses. The long-wavelength pulses, which overlapped the emission spectrum of pyridine2, resulted in a spatially localized decrease in intensity at the point of beam overlap. The time delayed quenching pulses caused a stepwise decrease in the intensity and anisotropy decays, as seen by oscillations in the frequency-domain data. The time resolved anisotropy was shown to decrease below zero (-0.2) following the vertically polarized quenching pulse. The extent of light quenching depended on the time delay between the excitation and quenching pulses, and can be used to measure the decay time. Light quenching and/or multipulse methods may provide a new class of experiments for fluorescence spectroscopy and imaging. PMID- 9203331 TI - Macromolecular crowding: effects on actin polymerisation. AB - Dextran has been found to enhance the polymerisation of actin. This enhancement increases exponentially with increasing mass concentrations of dextran, in a manner that is consistent with excluded volume theory. Mathematical prediction of experimental results is difficult due to the fact that all participating species, namely F-actin, G-actin and dextran are best represented by differently shaped hard particles. Modelling dextran as a sphere of radius defined by an effective thermodynamic radius (Reff), we have predicted our experimental results to an acceptable degree, given the relative crudity of the model. The results imply that the highly crowded cellular environment may help to stabilise the filamentous actin network in vivo. PMID- 9203332 TI - Solution conformation and dynamics of a tetrasaccharide related to the Lewix(X) antigen deduced by 1H NMR NOESY, ROESY, and T-ROESY measurements. AB - The conformational and dynamical features of a Le(X) tetrasaccharide analogue GalNAc (alpha 1-3)Gal(beta 1-4)[Fuc(alpha 1-3)]Glc(beta OMe) 1 have been studied through 1H NMR relaxation measurements. The results indicate that the different glycosidic linkages of 1 present distinct conformational flexibility in solution. In addition, the use of T-ROESY experiments in conformational analysis of oligosaccharides is explored emphasizing its scope and limitations. PMID- 9203333 TI - X-ray study of beijeran sodium salts, a new galacturonic acid-containing exo polysaccharide. AB - X-Ray fiber diffraction patterns were obtained from oriented films of sodium salts of a new uronic acid-containing polysaccharide (beijeran) both in its native, poly [-->3)-alpha-D-GalA-(1-->3)-beta-L-Rha-(1-->3)-alpha-D-Glc-O6Ac-(1 - >], and deacetylated forms. Initially the stretched films of both polysaccharides were amorphous, but the crystallinity was much improved by annealing at high temperature. The deacetylated specimen had higher crystallinity than the native. Both films showed similar X-ray fiber patterns indicating that these polysaccharides had similar unit cell dimensions and that the O-acetyl groups in the native beijeran chain did not disturb the regular array in the crystal having space group P21. All the visible reflections could be indexed in terms of a monoclinic unit cell with dimensions a = 1.277, b = 1.611, c (fiber axis) = 2.437 nm, and gamma = 96.79 degrees. The fiber axis length and the presence of (002) and (006) reflections indicated that the conformation was made up of two trisaccharide residues, in an extended two-fold helix. PMID- 9203334 TI - Motional properties of E. coli polysaccharide K5 in aqueous solution analyzed by NMR relaxation measurements. AB - 13C NMR relaxation measurements at three different magnetic field strengths have been used to analyse the motional properties of a low molecular weight K5 polysaccharide (delta UA-[-->4)-beta-D-GlcNAc(1-->4)-beta-D-GlcA(1-->]n GlcNAcred) from E. coli. Two-dimensional double INEPT spectra with suppression of cross-correlation effects between dipolar and chemical shift anisotropy relaxation mechanisms were collected in order to determine carbon longitudinal and transverse relaxation times. The values of the overall correlation time and the rate of internal motions were obtained using the model free spectral densities. The data indicate that the overall motion of the molecule is non isotropic and can be approximated with the symmetric top model with an axial ratio of approximately 22. The magnitude of the generalized order parameters (S2 approximately 0.8) and the internal motion correlation time (tau e approximately 30 ps) differ from those found for iduronic acid-containing glycosaminoglycans and suggest that the internal motions in K5 polysaccharide are more limited. PMID- 9203335 TI - 1H NMR relaxation study of a chitosan-cyclodextrin network. AB - The results of measurements of longitudinal and transverse proton relaxation times for a chemical network obtained by reacting chitosan with oxidized beta cyclodextrin (beta-cyclodextrin polyaldehyde) are presented. The network was characterized by a 'two-component' transverse relaxation mechanism relative to structurally different environments experienced by water molecules. Different environments were also indicated by the temperature of the spin-spin relaxation times (T2) studied in the range 4-50 degrees C. Between 4 and 18 degrees C, proton exchange between the matrix and water prevails on the inter- and intra molecular dipolar interactions of the water confined in the meshes of the network, resulting in a marked change in the slope of T2 with temperature. Stiffness of the matrix and reduced mobility of water in the gel meshes are prerequisites for observing such relaxation phenomena. Possible mechanisms contributing to the activation energy in the case of chitosan-cyclodextrin networks are discussed. The behaviour of the chitosan-cyclodextrin hydrogel is compared with that of a gellan gel. PMID- 9203336 TI - Oligosaccharides obtained by enzymatic hydrolysis of birch kraft pulp xylan: analysis by capillary zone electrophoresis and mass spectrometry. AB - Neutral and acidic oligosaccharides were obtained from an unbleached birch kraft pulp by treatment with a Trichoderma reesei endoxylanase pI 9 and subsequently characterized using capillary zone electrophoresis (CZE) and matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). The borate complexes of unsaturated acidic oligosaccharides having a 4-deoxy-beta-L threo-hex-4-enopyranosyluronic acid (4 delta UA) residue linked to a beta-D-(1- >4)-xylooligosaccharide backbone were separated by CZE and detected by their UV absorption at 232 nm without prior derivatization. Pre-column derivatization with the chromophore 6-aminoquinoline (6-AQ) followed by CZE in alkaline borate buffer using detection based on absorption at 245 nm was used in the case of neutral xylosaccharides. Furthermore, MALDI-TOF-MS was employed to determine the molecular masses of both unsaturated and saturated acidic oligosaccharides. The acidic oligosaccharides released upon endoxylanase treatment of the birch kraft pulp were a (4 delta UA)-beta-D-xylotetraose, a (4 delta UA)-beta-D-xylopentaose, a (4-O-methyl-alpha-D-glucurono)-beta-D-xylotetraose, and a (4-O-methyl-alpha-D glucurono)-beta-D-xylopentaose. Analysis after enzymatic hydrolysis with beta xylosidase and alpha-glucuronidase from Trichoderma reesei strongly indicated that the uronic acid residue in these acidic oligosaccharides was linked to the D xylose unit adjacent to the non-reducing D-xylose unit. The neutral xylosaccharides obtained after endoxylanase treatment of the pulp sample were D xylose, beta-(1-4)-D-xylobiose and beta-(1-4)-D-xylotriose. PMID- 9203337 TI - Chemoenzymatic synthesis of glucose fatty esters. AB - D-Glucose fatty esters at C-6 were obtained by chemoenzymatic synthesis involving 1,2-O-cyclohexylidene-alpha-D-glucofuranose (1) followed by hydrolysis of the cyclohexylidene protecting group. The enzymatic esterification of 1 was performed with fatty acids of variable chain lengths (C8:0 to C18:0). The kinetic of the reaction was studied for each fatty acid and the structure of the octanoyl ester was determined by 1H and 13C NMR spectroscopy. PMID- 9203338 TI - Chemical and chemo-enzymatic synthesis of the alpha-Neu p5Ac-(2-->6)- beta-D GalpNAc-(1-->4)-beta-D-GlcpNAc-(1-->2)-alpha-D-Manp element that is part of N linked carbohydrate chains of human lutropin. AB - In the framework of a project aimed at the elucidation of the nature of the functional importance of the N-glycosylation of the alpha-subunit of the glycoprotein hormones human lutropin and human chorionic gonadotropin, the structural element alpha-Neu p5Ac-(2-->6)-beta-D-GalpNac-(1-->4)- beta-D-GlcpNAc (1-->2)-alpha-D-Manp, which is part of the carbohydrate chains of human lutropin, has been prepared by chemical and chemo-enzymatic synthesis in the form of its propyl glycoside. Condensation of 4-O- acetyl-3,6-di-O-benzyl-2-deoxy-2 phthalimido-alpha/beta-D-glucopyranosyl trichloroacetimidate with allyl 3,4,6-tri O-benzyl-alpha-D-mannopyranoside gave after deacetylation allyl (3,6-di-O-benzyl 2-deoxy-2-phthalimido-beta-D-glucopyranosyl) -(1-->2)-3,4,6-tri-O-benzyl-alpha-D mannopyranoside. Ethyl 3-O-benzyl-2-deoxy-2-phthalimido-l-thio-beta-D glucopyranoside was converted into the galacto-derivative ethyl 4,6-di-O-acetyl-3 O-benzyl-2-deoxy-2-phthalimido-1-thio-beta-D -galactopyranoside via an oxidation reduction route, as well as via SN2-type substitution with acetate. The use of this galacto thioglycoside, after its conversion into the corresponding bromide, as GaIN donor for condensation with the mentioned disaccharide derivative yielded after deacetylation allyl (3-O-benzyl-2-deoxy-2-phthalimido-beta-D galactopyranosyl)-(1-->4) -(3,6-di-O-benzyl-2-deoxy-2-phthalimido-beta-D glucopyranosyl)-(1-->2) -3,4,6-tri-O-benzyl-alpha-D-mannopyranoside. Methylsulfenyl bromide-silver triflate promoted sialylation of this trisaccharide derivative with O-ethyl S-[methyl (5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy D -glycero-alpha-D-galacto-non-2-ulopyranosyl)onate] dithiocarbonate and subsequent deprotection resulted into the aimed tetrasaccharide structural element. Alternatively, this compound was prepared via a block synthesis, which, however, was not superior to the linear strategy. Finally, a stereose lective sialylation of synthetically prepared beta-D-GalpNAc-(1-->4)-beta-D-GlcpNAc-(1- >2)-alpha-D-Manp-(1-->O) CH2CH2CH3 with CMP-Neu5Ac and rat liver alpha-2,6 sialyltransferase was accomplished affording the same tetrasaccharide structural element. PMID- 9203339 TI - Structure of a new 6-deoxy-alpha-D-talan from Burkholderia (Pseudomonas) plantarii strain DSM 6535, which is different from the O-chain of the lipopolysaccharide. AB - An O-acetylated homopolysaccharide of 6-deoxy-D-talose (6-deoxy-alpha-D-talan polymer) was isolated from Burkholderia (Pseudomonas) plantarii strain DSM 6535 by extraction with 2-propanol. The structure (1) of the trisaccharide repeating unit of the polysaccharide was established by studies of the intact and O deacetylated polysaccharides using methanolysis, methylation analysis, 1H and 13C NMR spectroscopy, including 2D COSY, heteronuclear 13C, 1H COSY, 1D NOE, and computer-assisted analysis of 1D 13C NMR spectra. The remaining material after extraction of the biomass with 2-propanol showed to be a lipopolysaccharide with an O-specific polysaccharide chain having a different structure (2), which has been found previously in lipopolysaccharides of a number of other Gram-negative bacteria. [formula: see text] PMID- 9203340 TI - Specificity of amylases and cyclodextrin-glucanotransferase in reactions with 2 deoxy-maltooligosaccharides. AB - 2-Deoxy-maltooligosaccharides of different chain length were tested as substrates for exo- and endo-amylases. Cleavage occurred with beta-amylase, yielding 2,2' dideoxy-maltose, and with amyloglucosidase. With the alpha-amylase from Thermomonospora curvata tris-(2-deoxy)-maltotriose and the corresponding tetra- and pentasaccharides were formed. Porcine pancreatic alpha-amylase did not tolerate the deoxygenated substrate, nor were cyclization experiments with cyclodextrin-glucanotransferase (CGT) successful. In a coupling reaction with CGT, however, a series of transfer products to the acceptor 2-deoxyglucose were obtained. PMID- 9203341 TI - Porcine liver (2-->3)-alpha-sialyltransferase: substrate specificity studies and application of the immobilized enzyme to the synthesis of various sialylated oligosaccharide sequences. AB - In search of substrate analogues for the porcine liver beta-D-Gal p-(1-->3)-D-Gal p-NAc: CMP-Neu5Ac-(2-->3')-alpha-sialyltransferase, three disaccharides beta-D Gal p-(1-->3)-beta-D-Gal p-O-CH3 (5), beta-D-Gal p-(1-->3)-beta-D-(2-OAc)-Gal p-O CH3 (7) and beta-D-Gal p-(1-->3)-beta-D-(2-OAc)-Gal p-O-Bn (11) were synthesized and tested with the enzyme. Disaccharide 7 turned out to be a very good substrate allowing a rapid access to the trisaccharide alpha-Neu5Ac-(2-->3)-beta-D-Gal p-(1 ->3)-beta-D-(2-OAc)-Gal p-O-CH3 (13) on a preparative scale using the crude enzyme immobilized on cationic exchanger. Trisaccharide 13 was further exploited, first as a sialyl donor in Trypanosoma cruzi trans-sialidase catalyzed reaction and second through acetolysis reaction as a source for the synthon alpha-Neu5Ac (2-->3)-D-Gal (16). PMID- 9203342 TI - The measurement of affinity between ligand and antibody using ligand-induced antibody fluorescence change. PMID- 9203343 TI - Structure of the O-polysaccharide from the LPS of a Hafnia alvei strain isolated from a patient with suspect yersinosis. AB - The carbohydrate backbone of the Hafnia alvei strain Y166/91 lipopolysaccharide (LPS) was isolated by mild acid hydrolysis followed by gel permeation chromatography and studied by NMR spectroscopy and methylation analysis. Treatment with periodate and hypoiodite gave a modified polysaccharide which was also characterised. It was concluded that the polysaccharide has the following structure with two distinct regions, which are connected. The chain length parameters m and n were not determined but the ratio m/n is approximately 2. [- >3)-beta-D-Galp-1(1-->3)-alpha-D-Galp-(1-->]m[-->3)-alpha-D-Galp-(1-- >3)- beta-D Galf-(1-->]n From the present data it is not possible to determine whether it is the Galp-Galp chain or the Galp-Galf chain that is connected to the core. The structure found here is identical to that suggested for the O-specific polysaccharide chain of Klebsiella pneumoniae O1K2 (NCTC 5055) LPS [O. Kol, J.-M. Wieruszeski, G. Strecker, J. Montreuil, and B. Fournet, Carbohydr. Res., 217 (1991) 117-125; O. Kol, J.-M. Wieruszeski, G. Strecker, B. Fournet, R. Zalisz, and P. Smets, Carbohydr. Res., 236 (1992) 339-344]. PMID- 9203344 TI - Tobacco plants carrying a tms locus of Ti-plasmid origin and the Hl-1 allele are tumor prone. AB - The autonomous growth of plant tumor cells is believed to result from their persistent loss of the requirement for growth hormones such as auxin and cytokinin. The partially dominant gene Habituated leaf-1 (Hl-1) regulates the requirement of cultures tissues of Havana 425 tobacco (Nicotiana tabacum L.) for cytokinins. The Hl-1 allele can partially restore the tumor phenotype in tobacco cells transformed with a Agrobacterium tumefaciens Ti plasmid defective in the isopentenyl transferase locus, which encodes a key enzyme in cytokinin biosynthesis and is required for neoplastic growth. To investigate the oncogenic function of Hl-1, we transformed wild-type (hl-1/hl-1) and Hl-1/Hl-1 tobacco plants with the tms locus derived from the limited-host-range Ti plasmid pTiAg162. This locus encodes enzymes for biosynthesis of the auxin indole-3 acetic acid. Grafting tests and measurements of the hormone requirement of cultured explants show that wound-induced overgrowths arising in tms transformed Hl-1 plants are tumorous. While some wound-induced overgrowths also formed in hl 1/hl-1 transformants, these showed slight hormone-autotrophic growth and weak tumorigenicity in grafting tests. In addition, Hl-1/Hl-1 tms/tms plants, but not hl-1/hl-1 tms/tms plants, spontaneously developed rooty teratomatous overgrowths, showed flowering abnormalities, and formed calli at the base of the stem in young seedlings. Thus, Hl-1 tms plants exhibit a tumor-prone phenotype, and in this regard closely resemble tumor-prone hybrids that arise in certain interspecific crosses of Nicotiana species. Our results show that the interaction of just two genetic elements-the mutant Hl-1 allele of the tobacco host with tms genes of Ti plasmid origin-are sufficient for a tumor-prone phenotype. PMID- 9203345 TI - A marker of terminal stalk cell terminal differentiation in Dictyostelium. AB - We describe the isolation of staB, a Dictyostelium gene that is selectively expressed in stalk cells. If an aggregate enters culmination immediately staB is expressed in the bottom half of the stalk and in the basal disc but if a migratory slug is formed it is also expressed in the top of the stalk and in a disc of cells that surmounts the spore head. The latter two populations of cells derive from the rear part of the prestalk region, the pstO cells, which would therefore seem to change their transcriptional competency during slug migration. The staB promoter contains a distal region that is required for expression within both the stalk and the basal disc and a proximal region that is required only for expression within the basal disc. This uncoupling of transcriptional specificities suggests that the signalling mechanisms that direct terminal differentiation in these two tissues differ in some way. PMID- 9203346 TI - Inductive capacity of living eye tissues from adult frogs. AB - The aim of the present work has been to demonstrate the inducing capacity of living homogenous undamaged tissues from adult frogs. Tissues from the eyes of adult frogs Xenopus laevis: retina (R), pigmented epithelium (PE), lens epithelium (LE), and the forebrain (B) as a control, were tested for their inducing capacity using early gastrula ectoderm. To exclude the possibility of ad mixture of inducing cells in early gastrula ectoderm the transfilter induction technique was used throughout. The results show that the tissues used in the cases of most intense induction tended to induce similar cell types: both R and PE induce R + PE together with adjoining neural cells and secondary lens cells. LE induces lentoids (L) and B cells induce neuroids. In each case epidermis surrounds the explants filled with ectomesenchyme (EM) and melanophores (M). Immunofluorescence reactions clarified the nature of lens cells. This discovery indicates that cells of adult tissues continue throughout life producing substances which are able to promote the appearance of cells of the same type during development. Probably they also serve to mediate interrelations between cells of these tissues, regulating the stability of their differentiation in the adult state, as well. PMID- 9203347 TI - The intermediate filament protein nestin occurs transiently in differentiating testis of rat and mouse. AB - Nestin is an intermediate filament (IF) protein (IFP) which occurs during early developmental stages and during regenerative processes in muscle and neuronal cells. The spatial and temporal localization of nestin in the developing testis of rat and mouse was studied by immunolabeling light and electron microscopy and by immunoblotting. Nestin localization was related to the localization of the other major IFPs specific for this tissue, i.e. cytokeratins, vimentin and desmin. Laminin immunocytochemistry and conventional microscopy were used to identify tissues and cells. With the incipient differentiation of the gonadal anlage, the reaction for nestin was weak in the gonadal ridge, whereas the cells of the mesonephric mesenchyme showed a prominent reaction for this IFP. The nestin-specific reaction in the epithelial mesonephric duct and tubules was weak and disappeared at an early phase of differentiation. With the development of the testis proper, nestin was transiently found in several cell types. Nestin was found as well as vimentin and cytokeratins in the Sertoli cells. In the interstitial cells nestin was found together with vimentin and desmin IFPs, and was most prominent in the differentiating myoid cells. After birth, nestin gradually disappeared from the testicular cells and in the rat at puberty was found only in the endothelial cells of some blood vessels. The abolished nestin synthesis in the testis was confirmed by immunoblotting. These results suggest that nestin is required transiently during the development of the testis and mesonephros. The temporary presence of nestin, and several other IFPs during these phases, coincides with key phases of urogenital sex differentiation. This may imply that the orchestrated synthesis of the IFPs nestin, cytokeratins, vimentin and desmin is likely to be linked with the genes regulating sex differentiation. PMID- 9203348 TI - Stem cells of the corneal epithelium lack connexins and metabolite transfer capacity. AB - The stem cells of the corneal epithelial lineage are confined to the basal cell layer of the limbus, a vascularized outer corneal rim. These slow cycling cells of great proliferative potential maintain the corneal epithelial mass. Since cell cell communication plays an important role in development and differentiation, we conducted a comparative examination of the expression of two corneal connexins, C x 43 and C x 50, and the tracer transfer capacity of the limbal and corneal epithelia using the scrape loading technique. C x 43 is abundantly expressed in the basal cell layer of the epithelium covering the cornea, but is essentially absent from the mouse, human, neonatal rabbit, and chicken limbal epithelium. In the adult rabbit the limbal epithelium displays an overall weak C x 43 immunoreactivity, but C x 43-free isolated basal cells can be distinguished. C x 50 is expressed throughout the corneal epithelium of the three mammalian corneas, but is not detectable in the limbus. Scrape loading experiments in the rabbit yielded results which were consistent with the immunohistological findings; limbal epithelium lacked tracer (lucifer yellow) transfer capacity, strongly suggesting the absence of functional gap junctions. Altogether, our results demonstrate the incompetence of stem cells for gap junction-mediated cell-to-cell communication. This property may reflect the need of these unique cells to maintain a distinct intracellular environment. PMID- 9203349 TI - Formation of spheroid structures in a human colon carcinoma cell line involves a complex series of intercellular rearrangements. AB - The structural remodelling of tissues that occurs in vivo during animal morphogenesis can often prove difficult to study. Here we investigate the organizational processes of the LIM 1863 colon carcinoma cell line as it transforms from a single-cell stage into multicellular spherical structures called 'organoids'. The organoids can be dissociated into a viable single-cell suspension when cultured in calcium-depleted medium, and then induced to reform the organoid structure by the readdition of calcium. Previous studies have shown that initial cell attachment under these conditions is characterized by a novel mechanism of cell engulfment termed 'clutching'. This investigation reveals the subsequent appearance of junctional complexes between groups of 'clutched' cells prior to lumen formation, and the ultimate 'declutching' of entrapped cells as a means of cell rearrangement. Intact actin filaments but not microtubules were required for the initial clutching events, while inhibition of microtubule polymerization resulted in aberrant apical protein polarization, but did not affect the development of a luminal space within the spheroids. Single cells exhibited pools of intracellular microvilli contained in vacuolar apical compartments, which were resistant to the effects of cytoskeleton-disrupting drugs. However, these structures did not seem to be responsible for the swift development of the luminal surface observed in these cells. Two other cell lines, MDCK and DU 4475, were found to exhibit similar clutching conformations when induced to form three-dimensional structures, suggesting that this may be a widespread mechanism of cell rearrangement that reflects the process of organ morphogenesis in vivo. PMID- 9203351 TI - Patterns of postmating reproductive isolation in a newly discovered species pair, Aquarius remigis and A. remigoides (Hemiptera;Gerridae). AB - Aquarius remigoides Gallant and Fairbairn has recently been described as specifically distinct from A. remigis (Say) based upon genetic and morphological data. Both species are common, semiaquatic bugs (Hemiptera; Gerridae) found on the surface of streams and small rivers. Allozyme studies have shown them to be more distantly related than most other congeneric species in the Gerridae, with significant barriers to gene flow where their ranges abut. We assess postmating reproductive isolation between A. remigoides and A. remigis, using a bracket cross design with five sampling sites along a north-south cline traversing the hybrid zone. We also report the results of long-distance conspecific crosses of A. remigis, using populations from California and Quebec. Neither these nor the intraspecific bracket crosses reveal any evidence of isolation by distance within species. However, heterospecific crosses show significantly reduced fertility, hatching success, survival of both sexes to eclosion (final moult) and percentage of males. Egg production is not influenced by cross type, and we found no evidence of hybrid sterility in either sex. Analyses of genotypic frequencies at three isozyme loci in eight hybrid and 22 pure populations reveal significant deficiencies of heterozygotes in hybrid populations. The proportion of males is also significantly lower in hybrid populations than in pure populations. The laboratory and field results indicate that postmating isolation between these two species occurs in the form of severe reductions in both the fertility of heterospecific crosses and the viability of hybrids, particularly hybrid males. Genetic mechanisms for Haldane's rule and asymmetries in the reciprocal heterospecific crosses are discussed. PMID- 9203350 TI - Detection of Theileria annulata by the PCR in ticks (Acari:Ixodidae) collected from cattle in Mauritania. AB - We report on the detection of Theileria annulata in infected Hyalomma ticks by the PCR using primers derived from the gene encoding the 30 kDa major merozoite surface antigen (TamsI-1). No inhibition of the PCR was observed and as little as 0.1 pg of parasite DNA, corresponding to 12 sporozoites, could be detected in non infected tick DNA samples, spiked with T. annulata genomic DNA. Hyalomma dromedarii ticks, fed on a calf experimentally infected with T. annulata, were used to validate the PCR further. The infection rate in the adult ticks, fed as nymphs during the febrile reaction, was high (62%), dropped to zero for 1 day in tick batches that engorged after treatment with Butalex and increased to 30% 2 days later and 38% of the ticks acquired the infection after feeding as nymphs during a carrier state piroplasm parasitaemia of less than 0.1%. As an internal control, 16S tick rDNA sequences could be amplified from T. annulata-negative tick samples. Finally, 202 adult ticks from Mauritania, collected from zebu cattle carrying low levels of Theileria piroplasms, were tested by the PCR. Thirty-eight out of 52 (73%) and 17 out of 30 (57%) H. dromedarii from the Gorgol and Trarza regions, respectively and two out of 30 (7%) Hyalomma marginatum rufipes from the Gorgol region were positive. Hyalomma marginatum rufipes, Rhipicephalus evertsi evertsi and Rhipicephalus guilhoni from the Trarza region were negative. These findings confirm that H. dromedarii is the main vector of T. annulata in Mauritania and that the PCR is a useful method of determining the infection rates in ticks collected from cattle carrying low levels of T. annulata piroplasms. PMID- 9203353 TI - Significant genetic correlations among Caucasians at forensic DNA loci. AB - Although the effect of population differentiation on the forensic use of DNA profiles has been the subject of controversy for some years now, the debate has largely failed to focus on the genetical questions directly relevant to the forensic context. We re-analyse two published data sets and find that they convey much the same message for forensic inference, in contrast with the dramatically differing conclusions of the original authors. The analysis is likelihood-based and combines information across loci and across populations without assuming constant genetic differentiation. Our results suggest that the relevant genetic correlation coefficients are too large to be ignored in forensic work: although DNA profile evidence is typically very strong, the effect of genetic correlations can be important in some cases. Such correlations can, however, be accommodated in an appropriate assessment of evidential strength so that population genetic issues should not present a barrier to the efficient and fair use of DNA profile evidence. PMID- 9203352 TI - Analysis of the nucleolar organizing regions in the ant Tapinoma nigerrimum (Hymenoptera, Formicidae). AB - This study analyses the NORs of Tapinoma nigerrimum, a species that, as known from previous studies, has various chromosomes which carry a NOR site. The analysis was made by a combination of three methods: silver nitrate staining, in situ hybridization with fluorescein-or digoxigenin-labelled probes, and staining with the CG-specific fluorochrome chromomycin A3. The silver staining technique showed an Ag-positive region on chromosome 6 and on various other chromosomes. However, the application of in situ hybridization techniques showed only one positive signal in the proximal region of the short arm of chromosome 6 of T. nigerrimum. Similar results were observed by CMA banding. The absence of rDNA genes or the presence of only a small number of these, not detectable with the above probes, might explain the absence of hybridization signal in the remaining chromosomes. PMID- 9203355 TI - Cell model systems in plant cytoskeleton studies. AB - Cytoskeletons play an essential role in cellular functions in both animal and plant cells. In studies of the molecular mechanisms of their functions, a variety of cell model systems, mainly of animal cells, have yielded much information. With plant cells, cell model systems have mostly been restricted to studies on the mechanism of cytoplasmic streaming. Recently, however, there have been several reports of studies employing plant cell model systems to investigate plant cytoskeletons that have revealed new concepts about their structure and functions. To promote and support a general understanding of cell model systems, this review attempts to categorize them, present currently known information on the structure and function of plant cytoskeletons, and offer a possible role of cell model systems in future studies of plant cytoskeletons. PMID- 9203354 TI - Segregation studies and linkage analysis of Atlantic salmon microsatellites using haploid genetics. AB - A genetic marker map of Atlantic salmon would facilitate the identification of loci influencing economically important traits. In the present paper we describe five new Atlantic salmon microsatellites. Segregation studies and linkage analysis of these and previously published microsatellites were carried out in pedigrees consisting of diploid dams and haploid gynogenetic offspring. We confirm earlier reports that salmon microsatellites tend to have a higher number of repeat units than those of mammals. Linkage analysis revealed that three microsatellites belong to a linkage group spanning approximately 50 cM of the genome, whereas the remaining 10 markers seem to be unlinked. PMID- 9203356 TI - The structure, function, and assembly of actin filament bundles. AB - The cellular organization, function, and molecular composition of selected biological systems with prominent actin filament bundles are reviewed. An overall picture of the great variety of functions served by actin bundles emerges from this overview. A unifying theme is that the actin cross-linking proteins are conserved throughout the eukaryotic kingdom and yet assembled in a variety of combinations to produce actin bundles of differing functions. Mechanisms of actin bundle formation in vitro are considered illustrating the variety of physical and chemical driving forces in this exceedingly complex process. Our limited knowledge regarding the formation of actin filament bundles in vivo is contrasted with the elegant biophysical studies performed in vitro but nonetheless reveals that interactions with membranes, nucleation sites, and other organizational components must contribute to formation of actin bundles in vivo. PMID- 9203357 TI - Nuclear components with microtubule-organizing properties in multicellular eukaryotes: functional and evolutionary considerations. AB - The nucleus and the microtubular cytoskeleton of eukaryotic cells appear to be structurally and functionally interrelated. Together they constitute a "cell body". One of the most important components of this body is a primary microtubule organizing center (MTOC-I) located on or near the nuclear surface and composed of material that, in addition to constitutive centrosomal material, also comprises some nuclear matrix components. The MTOC-I shares a continuity with the mitotic spindle and, in animal cells, with the centrosome also. Secondary microtubule organizing centers (MTOC-IIs) are a special feature of walled plant cells and are found at the plasma membrane where they organize arrays of cortical MTs that are essential for ordered cell wall synthesis and hence for cellular morphogenesis. MTOC-IIs are held to be similar in origin to the MTOC-I, but their material has been translocated to the cell periphery, perhaps by MTs organized and radiating from the MTOC-I. Many intranuclear, matrix-related components have been identified to participate in MT organization during mitosis and cytokinesis; some of them also seem to be related to the condensation and decondensation of chromatin during the mitotic chromosome cycle. PMID- 9203358 TI - Sialic acids in molecular and cellular interactions. AB - Sialic acids (Sias) are terminal components of many glycoproteins and glycolipids especially of higher animals. In this exposed position they contribute significantly to the structural properties of these molecules, both in solution and on cell surfaces. Therefore, it is not surprising that Sias are important regulators of cellular and molecular interactions, in which they play a dual role. They can either mask recognition sites or serve as recognition determinants. Whereas the role of Sias in masking and in binding of pathogens to host cells has been documented over many years, their role in nonpathological cellular interaction has only been shown recently. The aim of this chapter is to summarize our knowledge about Sias in masking, for example, galactose residues, and to review the progress made during the past few years with respect to Sias as recognition determinants in the adhesion of pathogenic viruses, bacteria, and protozoa, and particularly as binding sites for endogenous cellular interaction molecules. Finally, perspectives for future research on these topics are discussed. PMID- 9203359 TI - Embryonic neural chimeras in the study of vertebrate brain and head development. AB - Construction of neural chimeras between quail and chick embryos has been employed since 1969 when the unique nucleolar structure of the quail nucleus and its use to devise a cell marking technique by associating quail and chick cells in ovo were described in the "Bulletin Biologique de la France et de la Belgique." This method was first applied to the ontogeny of the neural crest, a structure whose development involves extensive cell migration, and, since 1984, to that of the central nervous system (CNS). This chapter highlights some of the most significant findings provided by this approach concerning the CNS, such as (i) demonstration of the common origin of the floor plate and notochord from a group of cells localized in the "organizer", i.e., Hensen's node, and the way in which these two structures become positioned respectively within and under the neural tube during gastrulation and neurulation in Amniotes; (ii) the neural crest origin of the skull vault and the facial and hypobranchial skeleton. This means that the mesodermal contribution to the skull is limited to the occipital and otic regions and extends only to the rostral limit of the notochord. A correlation can be drawn between the development of the telencephalon and the mesectodermally derived skull in the vertebrate phylum; (iii) demonstration that the midbrain-hindbrain junction, at the stage of the encephalic vesicles, acts as an organizing center for tectal and cerebellar structures. This function was correlated with the activity of several developmental genes, thus providing insight into their function during neurogenesis; (iv) the pattern of morphogenetic movements and cell migration taking place in defined brain-to-be areas, as well as the origin of various cell types of nervous tissues; and (v) a new avenue for studying brain localization of either behavioral traits or genetically encoded brain disorders. PMID- 9203360 TI - Validation of ion chromatographic methods for the trace analysis of ions in pharmacopoeial grades of water. AB - Ion chromatography was investigated as an alternative technique to conventional pharmacopoeial methods for the determination of inorganic anions and cations in pharmacopoeial grades of water. These methods were validated by generating data on parameters such as specificity, linearity, precision, accuracy, sensitivity, ruggedness and stability of solution. The validation data demonstrate that ion chromatography is a viable alternative technique to current pharmacopoeial methods which enables automation of pharmacopoeial water analyses. PMID- 9203361 TI - Practical aspects of suppressed ion chromatography for cations in human serum. AB - In spite of the good accuracy and precision of ion chromatographic methods for the determination of mono- and divalent cations in human serum, the major drawback with these methods were problems with the membrane suppressor's performance. Here, we describe experiments undertaken to solve these problems. We address in particular the use of histidine-sulfuric acid eluents, sample purification with OnGuard-A cartridges and chromatographic "front-cut" for divalent cations. The latter two adaptations, resulting in removal of the anionic species from the sample, were successful in solving the observed suppressor problems. The eluent substitution, moreover, allowed us to switch from the chemical to the electric suppression mode. We believe that these adaptations will allow secure and robust determination of cations in human serum samples with ion chromatography. PMID- 9203362 TI - Determination of nitrite and nitrate in human serum. AB - A simple and effective assay for nitrite and nitrate in human serum has been developed using ion chromatography. Initial experiments using isocratic carbonate bicarbonate elution with conductivity detection on a Dionex QIC system with an AS4A-SC column showed promise but were unsatisfactory because of co-elution problems with nitrite. Carbonate and chloride were investigated as eluents using a gradient system, and direct UV detection at 214 nm was used in place of conductivity detection. Dionex AS4A, AS9A, AS12, Nucleopac PA-100 and Carbopac PA 100 columns were compared for selectivity and resistance to overload. The final method, using a chloride concentration gradient, pH buffering and direct UV detection with a Carbopac PA-100 column, shows good resolution, does not suffer from chloride overload and is simple to use. The method is being used in an investigation of the role of nitric oxide in pre-eclampsia, a hypertensive disorder during pregnancy. PMID- 9203363 TI - The role of nitric oxide in cancer. Improved methods for measurement of nitrite and nitrate by high-performance ion chromatography. AB - The short lifetime of nitric oxide (NO) in vivo impedes its quantitation directly; however, the determination of nitrite and nitrate ions as the end products of NO oxidation has proven a more practical approach. High-performance ion chromatographic analysis of nitrite in biological fluids is hampered by the large amount of chloride ion (up to approximately 100 mmol/l) which results in insufficient peak resolution when utilizing conductimetric detection. Analysis of both anions in small sample volumes is also constrained by the need to minimise sample handling to avoid contamination by environmental nitrate. We report a means to remove Cl- ions from small sample volumes using Ag+ resin which facilitates quantitation of either nitrite and nitrate anions in biological samples, using silica or polymer based ion-exchange resins with conductimetric or electrochemical and spectrophotometric detection. Including a reversed-phase guard column before the anion-exchange guard and analytical column also greatly extends column lifetime. PMID- 9203364 TI - Use of ion chromatography for monitoring microbial spoilage in the fruit juice industry. AB - Fruit juices and purees are defined as fermentable, but unfermented, products obtained by mechanical processing of fresh fruits. The presence of undesired metabolites derived from microbial growth can arise from the use of unsuitable fruit or from defects in the production line or subsequent contamination. This involves a loss in the overall quality that cannot be resolved by thermal treatment following the start of fermentation. With these considerations, together with microbiological control, the analysis of different metabolites, which can be considered as microbial growth markers, such as alcohols (i.e. ethanol, etc.), acids (i.e. acetic, fumaric, lactic, etc.) is fundamental in order to achieve a better evaluation of product quality. Enzymatic determination and other single-component analytical techniques are often used for the determination of these metabolites. When the microbial spoilage is not well known, this results in a long and cumbersome procedure. A versatile technique that is capable of determining many metabolites in one analysis could be helpful in improving routine quality control. For this purpose, an ion chromatographic technique, such as ion exclusion, for separation, and diode array spectrophotometry and conductivity, for detection, were evaluated. Both different industrial samples and inoculated samples were analyzed. PMID- 9203365 TI - In-line respeciation: an ion-exchange ion chromatographic method applied to the separation of degradation products of chemical warfare nerve agents in soil. AB - The natural background of anions encountered when analyzing soil samples by ion chromatography (IC) present significant problems in the separation, detection and quantification of isopropyl methylphosphonic acid (IMPA) and methylphosphonic acid (MPA), the degradation products of sarin, a chemical warfare nerve agent. Using chemically-suppressed IC with conductivity detection, a commercially available ion-exchange column, and an isocratic binary eluent system, IMPA and MPA were determined in aqueous extracts of soil at sub-ppm (microgram/g) concentrations without the need for gradient elution or organic solvent eluent modifiers. Common soil anions such as chloride, nitrate, sulfate and phosphate do not interfere with the analysis method due to the composition of the binary eluent allowing for greater mobilization of multivalent anions (e.g., MPA, carbonate, and sulfate) while monovalent anions (e.g., IMPA and nitrate) are relatively unaffected. Carbonate is selectively removed by in-line respeciation to bicarbonate. PMID- 9203366 TI - Determination of phosphate in natural waters by capillary electrophoresis. Influence of solution composition on migration time and response. AB - This paper deals with the determination of the more complex phosphate anion by capillary electrophoresis using indirect UV detection. First, the pH of the running electrolyte influences both the migration time and the response of the phosphate anion. Both effects could be explained well by taking into account phosphorus speciation in solution. In addition, the experimental method has been applied to three different sets of natural water systems; (i) groundwater, (ii) surface water and (iii) stemflow samples. The migration time behaviour of phosphate was different for the three sample sets and, hence, difficulties arise with respect to a clear and unique identification of the compound. Deviations herein could be minimized by applying a correction method for migration time drift. Concentrations of phosphate could be quantified in most samples and were confirmed by a colorimetric method. Average recoveries of additions of phosphate to groundwater, surface water and stemflow samples were 105, 83 and 103%, respectively. For one stemflow sample, quantitative recovery of phosphate was possible only by changing the pH of the running electrolyte solution. The latter observation might be very useful in setting up speciation-related measurement methods. PMID- 9203367 TI - Changes in selected aspects of immune function in the leopard frog, Rana pipiens, associated with exposure to cold. AB - The effect of exposure to low temperatures (5 degrees C) on lymphocyte proliferation, leukocyte populations, and serum complement levels was examined in the northern leopard frog, Rana pipiens. Proliferation of T lymphocytes in response to phytohemagglutinin stimulation was significantly decreased in frogs kept for 2, 3, and 5 months at 5 degrees C compared to that of animals kept at 22 degrees C. A significant increase in the average percentage of neutrophils and a decrease in the mean percentage of eosinophils was observed in the blood of frogs held for 5 months in the cold compared to animals held at 22 degrees C for the same length of time. Mean serum complement activity after 1 month at 5 degrees C was significantly reduced in comparison to animals held at 22 degrees C and was not detectable after 5 months in the cold. Recovery of complement levels at room temperature (22 degrees C) was also examined after cold exposure. Complement levels were significantly higher than controls (at 22 degrees C) in frogs returned to 22 degrees C for 7 and 14 days after 5 months in the cold. After frogs were held at 5 degrees C for 1 month, serum complement levels increased significantly within 2 days after returning to 22 degrees C and continued to rise 5 and 9 days after warming. Injections with Aeromonas hydrophila following a 5 week exposure to 5 degrees C failed to cause death or observable symptoms of disease in frogs that were returned to 22 degrees C. PMID- 9203368 TI - Timing of torpor bouts during hibernation in European hamsters (Cricetus cricetus L.). AB - Body temperature (Tb) of seven European hamsters maintained at constant ambient temperature (Ta = 8 degrees C) and constant photoperiod (LD 8:16) was recorded throughout the hibernating season using intraperitoneal temperature-sensitive HF transmitters. The animals spent about 30% of the hibernation season in hypothermia and 70% in inter-bout normothermy. Three types of hypothermia, namely deep hibernation bouts (DHBs), short hibernation bouts (SHBs), and short and shallow hibernation bouts (SSHBs), were distinguished by differences in bout duration and minimal body temperature (Tm). A gradual development of SSHBs from the diel minimum of Tb during normothermy could be seen in individual hamsters, suggesting a stepwise decrease of the homeostatic setpoint of Tb regulation during the early hibernation season. Entry into hibernation followed a 24-h rhythm occurring at preferred times of the day in all three types of hypothermia. DHBs and SHBs were initiated approximately 4 h before SSHBs, indicating a general difference in the physiological initiation of SSHBs on the one hand and DHBs and SHBs on the other. Arousals from SHBs and SSHBs also followed a 24-h rhythm, whereas spontaneous arousals from DHBs were widely scattered across day and night. Statistical analyses of bout length and the interval between arousals revealed evidence for a free-running circadian rhythm underlying the timing of arousals. The results clearly demonstrate that entries into hypothermia are linked to the light/dark-cycle. However, the role of the circadian system in the timing of arousals from DHBs remains unclear. PMID- 9203369 TI - Acute and chronic effects of temperature, and of nutritional state, on ion homeostasis and energy metabolism in teleost hepatocytes. AB - Short- and long-term effects of temperature on ion flux and energy turnover were studied in hepatocytes from thermally acclimated trout and roach. In trout hepatocytes K+ efflux was insensitive towards acute exposure to low temperature but was downregulated during cold acclimation of the fish so as to balance the uncompensated decreased K+ (Rb+) uptake of the cells. In contrast, both K+ (Rb+) uptake and K+ efflux of roach hepatocytes were temperature sensitive in the short term. These acute effects, however, were offset during cold acclimation by a near perfect compensation of both fluxes leading to re-establishment of ion flux homeostasis at the original level. Our findings, based on a new method permitting the simultaneous monitoring of K+ efflux and uptake in the same cell population, provide experimental verification of two of the three possible strategies, recently discussed by Cossins et al. (1995), by which the ionic steady state of fish cells may adjust to acute and chronic temperature change. By comparing hepatocytes from two groups of trout, one kept on a maintenance diet (ration I), the other fed ad libitum (ration II), we discovered striking effects of nutritional state on the absolute levels as well as on the temperature relationships of K+ uptake and protein synthetic activity. Both of these functions in the hepatocytes increased in the ration II fed as compared to the ration I fed trouts, but the increase of protein synthetic activity was greater and more uniform at the three experimental temperatures than that of K+ uptake. Moreover, protein synthetic activity proved to be considerably more temperature sensitive than K+ uptake and, in contrast to the latter, showed a compensatory response after cold acclimation. PMID- 9203370 TI - Second messenger and cAMP-dependent protein kinase responses to dehydration and anoxia stresses in frogs. AB - The effects of whole body dehydration (up to 40% of total body water lost) or anoxia exposure (up to 2 days under N2 gas) at 5 degrees C on tissue levels of adenosine 3'-5' cyclic monophosphate (cAMP) and the percentage of cAMP-dependent protein kinase present as the free catalytic subunit (PKAc), as well as the levels of the protein kinase C (PKC) second messenger, inositol 1,4,5 trisphosphate (IP3), were assessed in two anurans, the freeze-tolerant wood frog, Rana sylvatica, and the freeze-intolerant leopard frog, Rana pipiens. Dehydration of wood frogs resulted in a rapid elevation of liver cAMP and PKAc; cAMP was 3.4 fold greater than control values in animals that had lost 5% of total body water, whereas PKAc was elevated threefold in 20% dehydrated frogs. These results indicate protein kinase A mediation of the liver glycogenolysis and hyperglycemia that is induced by dehydration in this species. Skeletal muscle PKAc content also rose with dehydration but neither cAMP nor PKAc was affected by dehydration in leopard frog tissues. Anoxia exposure had different effects on signal transduction systems. PKAc was elevated after 1 h anoxia in R. sylvatica brain and was sustained over time but the enzyme was unaffected in other organs; by contrast, R. pipiens showed variable responses by PKAc to anoxia in three organs. Both species showed rapid (within 30 min) and large (3 to 7.8-fold) increases in IP3 in liver of anoxic frogs that decreased slowly with continued anoxia. IP3 also increased quickly in heart of anoxia-exposed wood frogs. This suggests that PKC may mediate various metabolic adjustments that promote hypoxia/anoxia resistance such as coordinating metabolic rate depression. A progressive rise in liver IP3 during dehydration in wood frogs (reaching fourfold higher than controls in 40% dehydrated animals) may also mediate similar hypoxia resistance adaptations under this stress since anurans experience progressive hypoxia due to increased blood viscosity when water loss reaches high values. The patterns of second messenger and PKAc changes in wood frog liver during dehydration closely parallel the changes seen in these same parameters during natural freezing suggesting that the freeze tolerance of selected terrestrially hibernating anurans may have evolved out of various anuran mechanisms of dehydration resistance. PMID- 9203371 TI - Biological effects of narrow-band (311 nm TL01) UVB irradiation: a review. AB - The narrow-band UVB (TL01) lamp (311 nm emission) was developed for use in phototherapy, as an alternative to a broad-band UVB source and to photochemotherapy, both of which have significant side effects and carry a risk of carcinogenesis. This new lamp has proved to be particularly effective at clearing psoriasis. It is now acknowledged that the TL01 lamp is probably 2-3 times more carcinogenic per minimum erythema dose than broad-band UVB, but the cumulative dose required in therapy is considerably less than when using broad band UVB sources. In terms of irradiation dose, the TL01 lamp is about 5-10-fold less potent than broad-band UVB for erythema induction, hyperplasia, oedema, sunburn cell formation and Langerhans cell depletion from skin. However, the broad-band UVB to TL01 potency ratio for cis-urocanic acid formation in the skin is approximately unity. In addition, the TL01 lamp, as used in phototherapy, has relatively more suppressive effects than broad-band UVB on systemic immune responses as judged by natural killer cell activity, lymphoproliferation and cytokine responses. However, the TL01 lamp is less effective at reducing epidermal antigen presentation, inducing dendritic cell migration to lymph nodes draining irradiated sites and suppressing contact hypersensitivity at the doses tested. Therefore the use of the TL01 lamp in phototherapy should be considered carefully after weighing up its diverse effects on the skin and immune system. PMID- 9203372 TI - The efficacy of an iron chelator (CP94) in increasing cellular protoporphyrin IX following intravesical 5-aminolaevulinic acid administration: an in vivo study. AB - 5-Aminolaevulinic acid (ALA)-induced protoporphyrin IX (PpIX) is proving to be a useful photosensitizer for photodynamic therapy (PDT). Conversion of PpIX to haem requires catalysed chelation with iron, and thus the presence of an iron chelator should, in theory, lead to an increase in cellular PpIX accumulation. This paper assesses the in vivo effect of a new iron chelator, 1,2-diethyl-3-hydroxypyridin 4-one (CP94), on the kinetics of PpIX in different layers of the bladder wall. Wistar rats were given 1% or 10% ALA intravesically with or without intraperitoneal CP94. The biodistribution of ALA-induced PpIX in the bladder was evaluated using fluorescence microscopy. Photodynamic effects on the bladder were compared in rats receiving various drug dosimetries. In CP94-treated rats, 5-7 h after administration of 10% ALA solution, the fluorescence intensity of PpIX in the urothelium was doubled compared with animals given ALA alone, whereas in the muscle layer PpIX remained at a low level similar to that found without the iron chelator. At an ALA concentration of 1%, although the PpIX concentration was not increased with CP94, the urothelial selectivity of PDT compared with the muscle layer was enhanced. In conclusion, by using CP94, a further reduction in skin photosensitization may be possible as similar photodynamic effects can be achieved with a lower dose of ALA. The addition of CP94 seems to be an effective and convenient way to potentiate ALA-induced PpIX tissue selectivity between the urothelium and the underlying layers of the bladder wall. PMID- 9203373 TI - Fluence rate as a determinant of synergistic interaction under simultaneous action of UV light and mild heat in Saccharomyces cerevisiae. AB - In experiments with wild-type diploid yeast cells of Saccharomyces cerevisiae, the synergistic lethal action of a simultaneous application of ultraviolet (UV) light (wavelength, 254 nm) and mild heat (45-57.5 degrees C) was studied. It was shown that, at any fixed UV light intensity, the synergistic effect occurred within the given temperature interval. The optimal temperature to achieve the greatest synergistic effect may be shown for every fluence rate examined. The correlation between the optimal temperature that maximized the synergy and UV light intensity was estimated: this temperature shifted towards higher temperature values with an increasing fluence rate. A possible interpretation of this effect is based on the supposition that the mechanism of the synergistic effect is related to additional lethal damage produced by the interaction of sublesions induced by each agent. These sublesions are supposed to be non-lethal when each agent is applied separately. PMID- 9203374 TI - Differential cytotoxic effects induced after photosensitization by hypericin. AB - The cytotoxic effects of the natural photosensitizing agent hypericin were evaluated. A dramatic difference in the sensitivity of several different human and mouse cell lines towards photoactivated hypericin (4 J cm-2) was demonstrated using a neutral red assay (e.g. A431, IC50 = 0.14 +/- 0.02 microM; HeLa, IC50 = 0.32 +/- 0.05 microM, MCF7, IC50 = 1.84 +/- 0.22 microM). Dark cytotoxicity was absent, even at high hypericin concentration (25 microM). The differential phototoxicity of hypericin did not correlate with the expression of the epidermal growth factor (EGF) receptor nor with the expression of the P170 glycoprotein in the cell. The reduction of the intracellular glutathione content did not enhance further the cytotoxic effects of photoactivated hypericin, as investigated with the A431, HeLa and MCF7 cells. Conversely, using confocal laser microscopy, it was shown that the phototoxicity correlated well with the hypericin cellular uptake. PMID- 9203375 TI - Cross-resistance to photofrin-mediated photodynamic therapy and UV light and recovery from photodynamic therapy damage in Rif-8A mouse fibrosarcoma cells measured using viral capacity. AB - Photodynamic therapy (PDT) utilizes the localized delivery of light to activate a photosensitizing drug (such as Photofrin) which is selectively retained by the tumour tissues. The intrinsic in vitro sensitivity of tumour cells to PDT is thought to be an important determinant of clinical tumour response to PDT. In this work we show the feasibility of using a viral capacity assay for adenovirus (Ad) DNA synthesis as an indicator of cellular sensitivity to and recovery from Photofrin-mediated PDT. Rif-1 mouse fibrosarcoma cells and a PDT resistant derivative, Rif-8A, as well as Chinese hamster ovary (CHO) cells and CHO-MDR multi-drug resistant mutant cells were studied. Consistent with the clonogenic survival of these cells, the capacity of PDT-treated cells for Ad DNA synthesis was greater for Rif-8A compared to Rif-1 cells and for CHO-MDR compared to CHO-N cells. Delaying infection of the Rif cells from immediately after, to 6 hours after PDT, resulted in an increased capacity for Ad DNA synthesis, which was greater for Rif-8A compared to Rif-1 cells, suggesting that the increased resistance of Rif-8A cells to PDT results from an elevated recovery and/or repair of PDT damage. The capacity of UV-irradiated cells for Ad DNA synthesis was also greater for Rif-8A compared to Rif-1 cells indicating a cross-resistance of Rif 8A cells to UV. These results suggest some overlap in the types of cellular damage induced by UV and PDT and/or overlap in the pathways for the repair of UV and PDT damage in Rif cells. PMID- 9203376 TI - Antioxidant adaptive response in human blood mononuclear cells exposed to UVB. AB - Human blood mononuclear cells exposed to UVB radiation develop increased antioxidant enzyme activities. Catalase (5.50 +/- 0.65 pmol (mg protein)-1), CuZn superoxide dismutase (16.7 +/- 2.1 pmol (mg protein)-1), Mn-superoxide dismutase (11.3 +/- 1.7 pmol (mg protein)-1), Se-dependent glutathione peroxidase (13.2 +/- 1.5 mU (mg protein)-1) and Se-independent glutathione peroxidase (3.30 +/- 0.52 mU (mg protein)-1) activities increase by 1.3-1.5-fold from the control activities after exposure to 0.3 W m-2 of 280-315 nm light for 15 min and a 3 h dark incubation period. DT-diaphorase activity (2.86 +/- 0.21 mumol DCPIP min-1 (mg protein)-1) increases threefold from the indicated control values. In contrast, cytochrome oxidase (0.36 +/- 0.04 min-1 (k') (mg protein)-1) and succinate dehydrogenase (3.06 +/- 0.25 mumol DCPIP min-1 (mg protein)-1) activities remain unchanged during the same irradiation and incubation period. The treatment of cells with cycloheximide prevents the response triggered by UVB exposure. These findings suggest that an inducible antioxidant defence mechanism operates on photo-oxidative stress and that both superoxide dismutase and DT diaphorase may display a concerted antioxidant role. PMID- 9203377 TI - In vivo fluorescence kinetics of porphyrins following intravesical instillation of 5-aminolaevulinic acid in normal and tumour-bearing rat bladders. AB - The fluorescence kinetics of protoporphyrin IX (PPIX) following intravesical instillation of 5-aminolaevulinic acid (5-ALA) have been studied in vivo in a rat bladder tumour model. 5-ALA dissolved in NaHCO3 was intravesically instilled for 60 min in tumour-bearing and normal bladders of Wistar rats. The fluorescence was excited with the violet lines of a Kr(+)-laser and recorded in vivo by means of a fibre coupled optical multichannel analyser. The fluorescence emission bands of PPIX at lambda = 636 nm and lambda = 708 nm were detected in normal and tumorous urothelium after only 30 min. The maximum fluorescence intensity was obtained in tumorous and normal urothelium 3-4 h after instillation. The ratio of the fluorescence intensity in tumorous to normal urothelium decreased continuously from four to about two during the time range of 6 h. PPIX fluorescence following 5-ALA instillation could also be observed in kidney and liver. Fluorescence from further porphyrin species with emission bands at lambda = 617 nm and lambda = 682 nm was detected in the bladder, indicating an efflux of hydrophilic porphyrins from the hepatic pathway. PMID- 9203378 TI - Indocyanine green: intracellular uptake and phototherapeutic effects in vitro. AB - Indocyanine green (ICG; absorption peak in human plasma 805 nm) was investigated for ICG-mediated phototherapy in vitro. The cellular uptake of ICG (1 microM-50 microM) into HaCaT keratinocytes after an incubation period of 24 h increased up to an intracellular ICG concentration of 12.1 +/- 1.3 nmol per 10(6) cells. To examine dose dependent phototoxic effects in vitro, keratinocytes were incubated with 0 microM-50 microM ICG for 24 h and irradiated by a diode laser (805 nm) with different energy densities (0, 12, 24, 48 J cm-2). All applied ICG concentrations except for 5 microM yielded a cell killing effect in combination with irradiation depending significantly on ICG concentration and light dose. Cell viability for dark control and cells incubated with 50 microM ICG and irradiated with 48 J cm-2 was 0.82 +/- 0.15 and 0.07 +/- 0.02, respectively. Sodium azide (100 mM), a quencher of reactive oxygen species, inhibited significantly the cell killing using 50 microM ICG and 24 J cm-2. Taken together, photoactivation of ICG by irradiation with a diode laser was shown to induce effectively cell killing of HaCaT keratinocytes. Moreover, this effect was inhibited by sodium azide, thus irradiation of ICG might induce a photodynamic reaction. PMID- 9203379 TI - Laser flash photolysis studies of electron transfer in complex III from yeast mitochondria. AB - The kinetics of reduction of cytochrome b and cytochrome c1 of yeast Complex III by 5-deazariboflavin semiquinone, generated by laser flash photolysis under anaerobic conditions, have been investigated. The reduction of cytochrome b occurs in two phases with first-order rate constants of 1300 and 670 s-1, whereas the reduction of cytochrome c1 appears as a unique exponential phase with an intermediate value of 800 s-1. Under these experimental conditions, about 50% of cytochrome b is reduced in comparison with cytochrome c1. After photoreduction, the re-oxidation of the cytochromes by internal re-equilibrium occurs in both cases, following pseudo-first-order kinetics at a rate constant of 43 s-1 for cytochrome b and 39 s-1 for cytochrome c1. These results, which agree with the data from the rapid mixing technique (A.-L. Tsai, J.S. Olson, G. Palmer, J. Biol. Chem. 262 (1987) 8677-8684), have implications for the mechanistic understanding of inner Complex III electron transfer. One of the goals of the investigation reported here is to provide direct evidence for the hypothesis of a proton-motive Q cycle for the mechanism of electron transfer in Complex III. Moreover, these results demonstrate the usefulness of laser flash photolysis in studying the redox kinetic properties of mitochondrial Complex III. PMID- 9203380 TI - Activity of 3-carbethoxyangelicin photolysis products. AB - 3-Carbethoxyangelicin (3-CA), carrying an electron-withdrawing group at the pyrone side, has been prepared to have a fully monofunctional angelicin derivative. 3-CA does not photoreact with DNA and induces a moderate antiproliferative activity. 3-CA proved to be extremely sensitive to ultraviolet A (UVA) light, undergoing rapid photolysis. Only one photolysis product has been isolated and identified. By means of alkaline elution, we observed that 3-CA and its photolysis products are able to induce a large amount of single-strand breaks in DNA in vivo. The results obtained from studying the capacity to produce singlet oxygen suggest that the photodynamic mechanism of action of 3-CA very likely results from its capacity--as well as that of its photolysis products--to produce singlet oxygen. PMID- 9203381 TI - Production of the free radicals O2.- and .OH by irradiation of the photosensitizer zinc(II) phthalocyanine. AB - Zinc(II) phthalocyanine (ZnPC) is a new photosensitizer currently undergoing phase I and II clinical trials at Lausanne's CHUV hospital for the photodynamic therapy (PDT) of early cancer in the upper aerodigestive tract. Activated oxygen species other than singlet oxygen produced during the photosensitization of ZnPC in liposomes have been examined by electron paramagnetic resonance (EPR) spin trapping and by the cytochrome c reduction method. Visible light irradiation of ZnPC associated with liposomes in the presence of the spin trap 5,5-dimethyl-1 pyrroline-1-oxide (DMPO) gives an EPR spectrum characteristic of the DMPO hydroperoxyl radical spin adduct (DMPO-.OOH). Superoxide anion attains a level of 1 microM min-1 20 min after the start of irradiation as determined by the superoxide dismutase (SOD)-inhibitable reduction of cytochrome c. The yield of O2.- is strongly enhanced by physiological electron donors. An EPR spectrum characteristic of the DMPO-hydroxyl radical spin adduct (DMPO-.OH) is also observed. The addition of dimethyl sulphoxide or ethanol produces additional hyperfine splittings due to the respective hydroxyalkyl radical products, indicating the presence of free .OH. DMPO-.OH is significantly inhibited by desferrioxamine or catalase. Conversely, this adduct is enhanced by hydrogen peroxide. These data demonstrate the ability of ZnPC in liposomes to photoreact effectively by an electron transfer mechanism. Such type I processes may add to the effects of singlet oxygen in ZnPC-mediated PDT. PMID- 9203382 TI - Singlet oxygen producing photosensitizers in photophoresis. AB - In this study, the immunosuppressive properties of two photosensitizers (benzoporphyrin derivative monoacid ring A (BPD) and Photofrin (HPD)), used for the photodynamic therapy of cancer, were investigated. The investigations were performed in our rat model for photophoresis. The validity of this model has been amply demonstrated. It enables a distinction to be made between antigen-specific and antigen non-specific immune suppression. With this model, the immune response which can be specifically suppressed is the contact hypersensitivity (CHS). CHS is induced by 2,4-dinitrofluorobenzene (DNFB). Both BPD and HPD are able to suppress CHS induced by DNFB. Furthermore, this generated suppression is transferable by the spleen cells of treated animals and is antigen non-specific. PMID- 9203383 TI - On-line fluorescence of human tissues after oral administration of 5 aminolevulinic acid. AB - It is important to have a frame of reference for the timing of photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA) so that PDT can occur when the tissue levels of protoporphyrin IX (PP) are at a maximum. This study describes a non-invasive fluorescence technique for detecting tissue PP levels after systemic ALA administration in patients with gastrointestinal cancer. The data suggest that the intensity of tumor surface fluorescence correlates with the tumor PP concentration. Spectrophotofluorometric measurements of skin and buccal mucosa also offer an easily acquired and rapid means for determining changes in plasma concentrations of PP. A number of potential variables, including blood flow, affect the intensity of fluorescence. We report that fluorescence measurements in situ are best adapted to the measurement of changes in the porphyrin levels in tissues rather than the absolute concentrations. PMID- 9203384 TI - Oxidative stress and in vivo chemiluminescence in mouse skin exposed to UVA radiation. AB - Mouse skin was exposed to UVA radiation (320-400 nm). The in vivo chemiluminescence of the skin was measured after irradiation. Chemiluminescence showed a maximum 13-fold increase (control emission, 10 +/- 1 cps cm-2) after 45 60 min of exposure to UVA, with no further increase with 60 min additional exposure. Spectral analysis of the emitted chemiluminescence showed that the principal species emitted in the 400-500 nm range. Topical application with alpha tocopherol (10% v/w) and beta-carotene (1 mM) greatly reduced the UVA-induced skin chemiluminescence. Thiobarbituric acid reactive substance (TBARS) levels were increased by 130% in skin homogenates after 2 h of exposure to UVA (control value, 77 +/- 14 nmol malonaldehyde equivalents (g tissue)-1). The activities of antioxidant enzymes in skin homogenates were decreased after 2 h of irradiation: the superoxide dismutase (SOD) activity (control value, 181 +/- 10 U SOD (g tissue)-1) was decreased by 40% and the catalase activity (control value, 1.34 +/ 0.14 pmol (g tissue)-1) was decreased by 45%. In vivo chemiluminescence appears to be a suitable method for following the kinetics of the oxidative stress processes and for testing the effect of topical application with antioxidant and photoprotective agents. PMID- 9203385 TI - 6,4,4'-Trimethylangelicin photoadduct immunodetection in DNA: induction and repair in Fanconi's anemia and normal human fibroblasts. AB - 6,4,4'-Trimethylangelicin (TMA)-photoinduced monoadducts (MAs) were detected and quantified on DNA of normal human and Fanconi's anemia (FA) fibroblasts (complementation groups A and D) by immuno-electron microscopy. TMA-modified DNA was extracted from the cells just after photoreaction, or after a subsequent 24 h repair period, for analysis of the MA processing capabilities of the different cell lines. Unmodified DNA was extracted from the control cells in parallel. The immunoreaction with antibody 7E3 was performed on single-stranded DNA fragments obtained by heat-formamide denaturation. On single-stranded DNA fragments scanned in the electron microscope, IgG-labeled MA sites appeared as isolated or clustered IgG molecules, which were not homogeneously distributed. The isolated IgG and the different clusters (doublets, triplets or near-neighbors (within a distance of 250 nucleotides)) were measured separately for induction frequency and removal. Few interstrand cross-links (CLs) were present on X-shape DNA fragments. At time zero, the distribution patterns of TMA-photoinduced IgG labeled MA sites and CLs, and their amount per 10(6) nucleotides, were similar in the three cell lines. After the 24 h repair period, FA cells from two different genetic complementation groups demonstrated impaired incision-excision repair capabilities for both MAs (singlets or clusters) and CLs when compared with normal cells. In each cell line, the relative proportions of TMA-induced lesions remaining at time 24 h were similar to those initially induced. This implies analogous processing kinetics towards the TMA-photoinduced clusters of MAs and CLs in a given cell line. PMID- 9203386 TI - Photopheresis, a possible therapy for airway hyperreactivity? AB - Airway hyperreactivity is an almost universal feature of asthma. The exact origin of this phenomenon is poorly understood. However, there is increasing evidence that T cells play an important role in the pathogenesis of this disorder. This fact makes it challenging to photopheresis to suppress the pulmonar hyperreactivity response. Photopheresis is a therapy for T cell mediated diseases aiming at specific suppression of the pathogenic clone of T cells involved. The use of photopheresis for the treatment of airway hyperreactivity was investigated in this study. We performed experiments in a murine model for airway hyperreactivity. In short, mice were sensitized by cutaneous application of 2,4,6 trinitrochlorobenzene. The immune system was challenged by an intratracheal injection of 2,4,6-trinitrobenzenesulfonic acid and a bronchoalveolar lavage was performed. In this lavage the total number of leukocytes was established and the number of macrophages was determined. It was found that photopheresis treatment was capable to suppress the airway hyperreactivity response for about 80%. In addition, the generated suppression proved to be transferable by splenocytes of treated animals. We conclude that photopheresis can be an interesting therapy for airway hyperreactivity (and perhaps also for asthma) especially when one takes into account that photopheresis induces specific immune suppression and has hardly any adverse effects. PMID- 9203387 TI - Reconstruction of in vivo skin autofluorescence spectrum from microscopic properties by Monte Carlo simulation. AB - The in vivo skin autofluorescence spectrum was reconstructed by Monte Carlo simulation using microscopic fluorophore distributions and intrinsic fluorescence spectra measured from excised skin tissue sections as well as employing published skin tissue optical parameters. The theoretical modeling took into account the light-tissue interactions of scattering, absorption, and regeneration of fluorescence photons. The modification of the intrinsic spectra by tissue optical properties to generate the in vivo spectrum observed at the tissue surface can be represented by a fluorescence detection efficiency function (eta) which equals the integral of the product of the excitation light distribution inside the tissue and the fluorescence escape efficiency. Comparison of the reconstructed in vivo spectrum with the measured spectra showed good agreement, outside of the blood absorption bands, suggesting that (i) the theoretical modeling, (ii) the skin optical parameters used, and (iii) the measured microscopic morphology and spectral data are consistent. The divergence which exists over the strong blood absorption wavelength band (530-600 nm) suggests that the effect of blood contents on in vivo tissue optical properties deserves further investigations. PMID- 9203388 TI - Does the in-vitro efficiency of meso-tetrahydroxy-phenyl-chlorin depend on pre treatment of sensitizer? AB - The efficiency of a new photosensitizer of the second generation, meso-tetra hydroxyphenyl-chlorin (mTHPC), which has a strong absorption at 652 nm, was investigated by oxygen consumption measurements and membrane integrity testing. The experiments proved a great increase in the efficiency of mTHPC after preincubation at 37 degrees C for 24 hours. From these findings it can be assumed that tumor cells can be treated in an optimal way with PDT after a longer delay following drug administration. PMID- 9203389 TI - PDT light dosimetry revisited. AB - A versatile PDT light dosimetry model is described incorporating the effects of drug photobleaching, drug elimination, and normal tissue damage. The dependence of the necrosis depth (dn) on the incident light dose for the four major modes of PDT light delivery has the form: dn = delta loge(DG), where delta is the optical penetration depth of the tumor tissue, D is the ratio of the incident light dose to the energy fluence at the necrosis threshold, and G is a function of the tissue optical constants. Light dosimetry graphs were calculated for Photofrin at standard conditions. PMID- 9203390 TI - The importance of fluorescence distribution and kinetics of ALA-induced PpIX in the bladder in photodynamic therapy. AB - Photodynamic therapy (PDT) is a new anticancer technique directed at the selective destruction of neoplastic tissue. Aminolevulinic acid (ALA), a precursor in the biosynthesis of home, induces the production of the endogenous photosensitizer protoporphyrin IX (PpIX). In this study the fluorescence distribution of ALA-induced PpIX was investigated in the rat bladder wall by fluorescence microscopy. The fluorescence studies showed that PpIX concentrates in bladder mucosa and that the highest fluorescence levels are achieved after four hours of 300 mg/kg ALA administration. A clear trend in difference between mucosa and muscularis layers was found in all samples taken after 1, 2, 4 and 6 hours of ALA administration. Our results suggest that to get the highest PpIX fluorescence intensity in bladder mucosa it is best to use 300 mg/kg ALA administration. Four hours is the time point when the highest difference in fluorescence between mucosa and muscularis is reached. PMID- 9203391 TI - Inhibition of pteridine biosynthesis eliminates blue-light dependent stimulation of red-light saturated photosynthesis in Laminaria saccharina (L.) Lamouroux. AB - Blue-light stimulation of red light-saturated photosynthetic oxygen evolution in Laminaria saccharina (L.) Lamouroux could be abolished within 5 days by incubation in a solution of 2,4-diamino-6-hydroxypyrimidine (DAHP), an inhibitor of GTP-cyclohydrolase I (E.C.3.5.4.16) activity. Photosynthesis in red light was not detectably affected. GTP-cyclohydrolase I, which catalyses the first step in the biosynthetic pathway of pteridines, was shown to be active in Laminaria. Under conditions that lead to complete inhibition of the photosynthetic stimulation, DAHP reduced the content of the pteridines in the tissue considerably. The amount of pterin was about 14%, that of biopterin was about 45% and that of an unidentified pteridine was about 27% of those of the controls. By contrast, the total concentration of flavins (FAD + FMN - riboflavin) was not significantly affected. The results suggest that pterins may be involved in the response of photosynthesis to blue light, possibly participating in photoreception. PMID- 9203392 TI - A medieval controversy about odor. AB - Medieval scholars debated whether odor was transmitted through a medium as suggested by Aristotle or by fumes or vapors from the odoriferous object as suggested by Plato. Key evidence believed to support Aristotle's theory was the behavior of birds in detecting carrion from far away. The medieval approach to this essentially secular controversy was, nevertheless, similar to that used in issues thinkers of that time regarded as of more importance. PMID- 9203393 TI - Second-order motion perception in the peripheral visual field. AB - In motion perception, luminance-defined stimuli (first-order motion) are distinguished from stimuli defined by more complex attributes (second-order motion), because they differ in their processing requirements. For instance, a two-layer model with the output of an array of elementary motion detectors (EMD's) feeding into a second array of EMD's has been proposed to account for seeing the movement of motion-defined objects. The question is raised whether this processing scheme is operating across the whole visual field or whether second-order motion perception is restricted to the fovea. The detection, orientation discrimination, and motion direction discrimination of oblique, vertically moving bars was tested at horizontal eccentricities between 0 degree and 16 degrees. Bars were defined on a dynamic noise background by an area of static dots (drift-balanced motion) or by coherent dot motion either in the direction of the bar motion (Fourier motion) or in the orthogonal direction (theta motion). Coherence thresholds for direction discrimination are severely impaired in the periphery for both types of second-order motion but not for Fourier motion, whereas orientation discrimination and detection marginally decline for all three bar types when the stimuli are presented further out in the periphery. In a control experiment it is shown that this result cannot be due entirely to the changes in spatial scale of the peripheral visual system. The facts that motion-defined objects can be detected in the periphery and that their orientation can be detected, but not their direction of motion, supports the view that the two-layer system suggested for the processing of theta motion is restricted to the central region of the visual field. PMID- 9203394 TI - Bayesian color constancy. AB - The problem of color constancy may be solved if we can recover the physical properties of illuminants and surfaces from photosensor responses. We consider this problem within the framework of Bayesian decision theory. First, we model the relation among illuminants, surfaces, and photosensor responses. Second, we construct prior distributions that describe the probability that particular illuminants and surfaces exist in the world. Given a set of photosensor responses, we can then use Bayes's rule to compute the posterior distribution for the illuminants and the surfaces in the scene. There are two widely used methods for obtaining a single best estimate from a posterior distribution. These are maximum a posteriori (MAP) and minimum mean-square-error (MMSE) estimation. We argue that neither is appropriate for perception problems. We describe a new estimator, which we call the maximum local mass (MLM) estimate, that integrates local probability density. The new method uses an optimality criterion that is appropriate for perception tasks: It finds the most probable approximately correct answer. For the case of low observation noise, we provide an efficient approximation. We develop the MLM estimator for the color-constancy problem in which flat matte surfaces are uniformly illuminated. In simulations we show that the MLM method performs better than the MAP estimator and better than a number of standard color-constancy algorithms. We note conditions under which even the optimal estimator produces poor estimates: when the spectral properties of the surfaces in the scene are biased. PMID- 9203395 TI - Improved microscopic identification of Clavibacter michiganensis subsp. sepedonicus cells by combining in situ hybridization with immunofluorescence. AB - An oligonucleotide probe was selected from the 16S rRNA gene of Clavibacter michiganensis subsp. sepedonicus for specific in situ hybridization. The rhodamine-labelled oligonucleotide probe was used in conjunction with an indirect immunofluorescence procedure based on a specific monoclonal antibody detected with a fluorescein-labelled conjugate. Simultaneous labelling of bacterial cells with the oligonucleotide and antibody probes allows accurate microscopic identification of single cells when isolation and other methods of confirming bacterial identity are not possible. PMID- 9203396 TI - Co-metabolism and microbial growth in the biodegradation of alkylbenzenesulphonates. AB - Two organisms, CCMI507 and CCMI852, degrading undecylbenzenesulphonate (LAS) by the ortho- and meta-cleavage pathways were studied in cultures where glucose was used as carbon and energy source. CCMI507 (ortho-pathway) started the degradation of LAS at the beginning of the culture development in parallel with glucose utilization. The degradation followed a steady profile of degradation until 77% of LAS was degraded in the culture containing initially 5 mg l-1 of the compound and 81% in the cultures containing initially 10 and 20 mg l-1 of LAS, after 72 h fermentation. The organism CCMI852 (meta-pathway) started degrading the compound only after 20 h, when 75% of glucose was spent and well within the stationary phase. After 72 h fermentation the level of degradation by CCMI852 varied from 70% (5 mg l-1 of LAS) to around 75% (10 and 20 mg l-1 of LAS). PMID- 9203398 TI - Degradation of 3-chlorobenzoate by thermophilic micro-organisms. AB - Thermophilic (75 degrees C), anaerobic biodegradation of chlorobenzoates was investigated using different inocula from geothermal and non-geothermal environments. Microbial dehalogenation of 3-chlorobenzoate (0.5 mmol l-1 was achieved by two mixed cultures growing anaerobically at 75 degrees C. One culture consisted of a facultative anaerobe and two obligate anaerobes, one of which was a methanogen, isolated from terrestrial sediments from hot springs in New Zealand. The other culture, derived from a non-geothermal environment, consisted of a Clostridium spp. and a non-spore-forming obligate anaerobe. No degradation of either 2-chlorobenzoate or 4-chlorobenzoate was achieved by these thermophilic cultures over the same time period. This is the first reported biotransformation of this chlorinated aromatic at a temperature of 75 degrees C. PMID- 9203397 TI - A high homology exists in N-terminal amino acid sequences of delta-endotoxins between Lepidoptera-specific and Coleoptera-specific Bacillus thuringiensis strains. AB - The 130 kDa parasporal inclusion proteins of the Lepidoptera-specific Bacillus thuringiensis reference strains for four H serovars were examined for sequences of 14 N-terminal amino acids. The four sequences fell into a single category, MNRNNQNEYEVIDA, and were dissimilar to any of the established sequences for Lepidoptera- and/or Diptera-killing crystal proteins. They were highly related to the reported sequence of the 130 kDa protein of the strain Buibui (serovar japonensis), which is specific for scarabaeid coleopterans. PMID- 9203399 TI - A comparison of immunomagnetic and surface adhesion immunofluorescent techniques for the rapid detection of Listeria monocytogenes and Listeria innocua in meat. AB - An immunomagnetic immunofluorescent method was investigated for the rapid detection of Listeria monocytogenes and Listeria innouca. This technique involved enrichment of the suspect sample at 30 degrees C overnight. Listeria monocytogenes cells were isolated from the enriched sample using immunomagnetic separation and Listeria were subsequently visualized using an immunofluorescent microscopy technique. This technique was used in the detection of Listeria cells from pure culture, inoculated beef mince samples and naturally contaminated retail beef mince samples. A detection level of approximately 1 x 10(3) cfu ml-1 was achieved. When compared with traditional detection methods no false negatives or positives were recorded for L. monocytogenes or L. innocua. The immunomagnetic immunofluorescent technique had a detection level similar to a previously described surface adhesion immunofluorescent technique. Isolation of the Listeria cells by surface adhesion involved dipping a membrane attached to a microscope slide into the enriched sample for 10 min. This was quicker and simpler to perform than the immunomagnetic separation technique which took 2 h to carry out. PMID- 9203402 TI - A novel tubular bioassay for measuring the production of antagonistic chemicals produced at the fungal/pathogen interface. PMID- 9203401 TI - Evaluation of the Biolog system for the identification of food and beverage yeasts. AB - The inconvenience of conventional yeast identification methods has resulted in the development of rapid, commercial systems, mainly for clinical yeast species. The Biolog system (Biolog Inc., Hayward, CA, USA) is a new semi-automated, computer-linked technology for rapid identification of clinical and non-clinical yeasts. The system is based around a microtitre tray and includes assimilation and oxidation tests. This paper evaluates the Biolog system for the identification of 21 species (72 strains) of yeasts of food and wine origin. Species correctly identified included Saccharomyces cerevisiae, Debaryomyces hansenii, Yarrowia lipolytica, Kluyveromyces marxianus, Kloeckera apiculata, Dekkera bruxellensis and Schizosaccharomyces pombe. Zygosaccharomyces bailii and Zygosaccharomyces rouxii were identified correctly 50% of the time and Pichia membranaefaciens 20% of the time. PMID- 9203404 TI - Antibiotic resistance and plasmid profile of Aeromonas hydrophila isolates from cultured fish, Telapia (Telapia mossambica). AB - Strains of Aeromonas hydrophila isolates from skin lesions of the common freshwater fish, Telapia mossambica, were screened for the presence of plasmid DNA by agarose gel electrophoresis and tested for susceptibility to 10 antimicrobial agents. Of the 21 fish isolates examined, all were resistant to ampicillin and sensitive to gentamycin. Most isolates were resistant to streptomycin (57%), tetracycline (48%) and erythromycin (43%). While seven of 21 isolates harboured plasmids, with sizes ranging from 3 to 63.4 kilobase pair (kb), it was only possible to associate the presence of a plasmid with antibiotic resistance (ampicillin and tetracycline) in strain AH11. Both the plasmid and the associated antimicrobial resistance could be transferred to an Escherichia coli recipient by single-step conjugation at a frequency of 4.3 x 10(-3) transconjugants per donor cell. PMID- 9203405 TI - Generation of polyclonal antibodies against a chemically synthesized N-terminal fragment of the bacteriocin pediocin PA-1. AB - Six mice were immunized intraperitoneally (i.p.) with a chemically synthesized 9 mer fragment (PH1) designed from the N-terminal part of the bacteriocin pediocin PA-1 and conjugated to keyhole limpet haemocyanin (KLH). After three doses of the immunogen had been administered, serum-specific antibodies were detected by a competitive direct ELISA. Myeloma cells were injected i.p. into mice in order to obtain ascites polyclonal antibodies. Although four mice developed ascites, only mouse 2 had detectable specific antibodies in the ascites fluid. The serum and ascites antibodies were specific for PH1 but they did not recognize the whole pediocin PA-1 molecule. This is the first attempt to generate antibodies against bacteriocins with a chemically synthesized oligopeptide as immunogen. This approach still remains attractive for detection, quantification, mode of action studies and purification of bacteriocins, especially those for which the purification process is difficult or inefficient at present. PMID- 9203406 TI - Immunomagnetic separation and conventional culture procedure for detection of naturally occurring Salmonella in raw pork sausages and chicken meat. AB - The aim of the study was to compare immunomagnetic separation (IMS) and conventional selective enrichment procedures using selenite cystine broth (SC) and Rappaport-Vassiliadis broth (RV) in 137 naturally contaminated food samples (69 raw pork sausages and 68 chicken meat). The utilization of SC or IMS appeared to be the most appropriate enrichment procedure: 15 out of 18 Salmonella-positive samples (83.3%) were detected by SC and 12 (66.7%) by IMS; RV yielded only seven positive isolations (38.9%). However, RV yielded the highest count of Salmonella colonies per plate and the lowest interference by competing organisms. IMS could become a reliable alternative to standard enrichment procedures and a combined IMS and selective enrichment broth could increase the chance of Salmonella recovery. PMID- 9203407 TI - Efficiency of the polymerase chain reaction amplification of the uid gene for detection of Escherichia coli in contaminated water. AB - Direct detection of Escherichia coli from polluted river water was achieved using polymerase chain reaction (PCR) amplification of the uid gene. Amplification using DNA from environmental samples resulted in non-specific DNA fragments. Specific amplification was achieved through use of the touch-down PCR procedure. Targeting the uidA structural region of the gene gave reproducibly better amplification than targeting the uidR regulatory region. The data demonstrate conditions for optimal specific detection. PMID- 9203408 TI - T cell clonal anergy. AB - Recent experiments have elucidated two molecular mechanisms that may account for the failure of anergic T cell clones to initiate IL-2 gene transcription following TCR stimulation. First, a block has been identified in the ERK and JNK mitogen-activated protein kinase pathways; the block results from a failure to activate p21ras. It leads to reduced induction of c-Fos and JunB proteins and to a failure to form and phosphorylate the activator protein (AP)-1 heterodimers required for IL-2 gene transcriptional activation. Second, repressor molecules (Nil-2-a and a molecule related to AP-1) have been characterized that dominantly inhibit IL-2 gene transcription. PMID- 9203409 TI - Pathways leading to cell death in T cells. AB - Antigen-induced apoptosis of T cells is a highly regulated process which plays a key role in the elimination of self-reactive T cells and, thus, in the prevention of autoimmunity. It has recently become apparent that members of the tumor necrosis factor (TNF) and TNF receptor (TNFR) superfamily regulate antigen induced T-cell death. Studies characterizing genes which control TNF/TNFR superfamily expression and how TNF/TNFR signal transducers activate cell death machinery, such as caspases, have begun to reveal the molecular control of antigen-induced T-cell death. PMID- 9203410 TI - Cell death in the regulation of immune responses. AB - Cell death is an integral part of the functioning of the immune system. For T cells, potentially autoreactive or 'useless' cells are removed through apoptosis in response to signals (or lack of signals) from their T cell receptor complex. A myriad of proteins that can initiate or protect cells from cell death have recently been identified. PMID- 9203411 TI - Immunological memory. AB - Most of the antigen-specific T and B cells participating in the primary immune response are rapidly eliminated, but some of the cells survive and become long lived memory cells. There have been a number of recent developments on the features and functions of memory cells. PMID- 9203412 TI - Xid and Xid-like immunodeficiencies from a signaling point of view. AB - The analysis of Btk-associated molecules and ligand-induced Btk phosphorylation has suggested the existence of a complexed Btk-associated signaling network involved in the activation of B lymphocytes and mast cells. Recent gene targeting experiments have revealed protein kinase C betaI/II (PKCbetaI/II) as a critical component of the Btk-dependent signaling chain and have highlighted a potential role for the Btk-PKCbetaI/II interaction in the amplification of B cell receptor mediated signaling. PMID- 9203413 TI - Co-receptors of B lymphocytes. AB - The past year has seen advances in our understanding of accessory membrane proteins that modulate the B cell response to antigen-receptor stimulation. The generation of complement receptor deficient mice has reinforced our appreciation of the importance of complement receptors in the B cell response to antigen. The association of inositol polyphosphate 5-phosphatase with FcgammaRIIB suggests another mechanism, in addition to recruitment of the phosphotyrosine phosphatase SHP-1, by which secreted immunoglobulin can limit further response to antigen. The in vivo function of CD22 in regulating the threshold of antigen-receptor signalling has been shown using CD22-deficient mice. Lastly, B cell receptor signalling in the B-1 subset of B lymphocytes has been demonstrated to be negatively regulated by CD5. PMID- 9203414 TI - Inhibitory receptors, ITIM sequences and phosphatases. AB - A diverse group of inhibitory receptors, including FcgammaRII, killer cell inhibitory receptors, and B22, shares an immunoreceptor tyrosine-based inhibition motif (ITIM). Recent studies have shown that this motif, when phosphorylated on tyrosine, forms a docking site for the Src homology 2 recognition domains of the protein tyrosine phosphatase SHP-1 and the inositol 5-phosphatase SHIP. A similar motif in cytotoxic T-lymphocyte antigen-4 recruits the related tyrosine phosphatase SHP-2. These three enzymes act to inhibit signaling cascades resulting from ligation of the BCR, TCR, FcgammaRIII, and FcepsilonRI, although the relative importance of the tyrosine phosphatases and the inositol phosphatase differs depending on the cell type. PMID- 9203415 TI - Molecular mechanisms in B cell antigen receptor signaling. AB - Recent gene-targeting experiments have highlighted the importance of the intracellular protein tyrosine kinases Lyn, Syk, and Btk in BCR signal transduction and B cell development. In addition, the interactions of these kinases and their regulatory mechanisms have been reported. Activation loop phosphorylation of these kinases is critical for their participation in signal propagation. Several substrates have been identified for these kinases and this has led to elucidation of the mechanisms by which these kinases mediate the downstream signaling events that lead to cellular responses of B cells. PMID- 9203416 TI - TCR zeta chain in T cell development and selection. AB - Current data suggest that an important function of the multimeric structure of the TCR is to enable the assembly of structurally and functionally different forms of the TCR, the pre-TCR and alphabetaTCR complexes, at different stages in development. Four distinct TCR subunits (the CD3gamma, delta, and epsilon chains and the zeta chain) contain signal transducing motifs; however, the zeta chain is notable for containing three of these elements. These motifs, especially those within the zeta chain, function to amplify signals generated by the TCR, and this property is especially critical during thymocyte selection. The results of several recent experiments argue that positive and negative selection of thymocytes may involve activation of distinct downstream signaling pathways. The outcome of thymocyte selection can also be influenced, however, by quantitative effects such as changes in ligand concentration or direct alteration of the TCR signaling potential. Recent studies pertaining to the kinetics of TCR-ligand interactions may provide insight into how signaling through the TCR can be regulated either quantitatively or qualitatively. PMID- 9203417 TI - Natural killer cell receptors. AB - The specificity in the recognition of hematopoietic target cells by natural killer cells is primarily provided by inhibitory receptors and several such receptors have been identified in the past year. Surprisingly, the recognition of MHC class I molecules by inhibitory receptors on human natural killer cells involves two different types of receptors, one with Ig domains (killer cell inhibitory receptor), and another with C-type lectin domains (CD94-NKG2). Mouse natural killer cells recognize MHC class I molecules through the C-type lectin Ly49 receptors but also express a receptor, of unknown ligand specificity, that is related to the killer cell inhibitory receptor. PMID- 9203418 TI - Functions of CD40 on B cells, dendritic cells and other cells. AB - CD40 is a cell surface receptor that belongs to the tumor necrosis factor receptor family. It was first identified and functionally characterized on B lymphocytes; however, in recent years it has become clear that CD40 expression is much broader, as it is found on monocytes, dendritic cells, hematopoietic progenitors, endothelial cells and epithelial cells. Although initially identified for its activation properties, CD40 is also able to transduce negative signals in various cell types. It is presently accepted that CD40 plays a critical role in the regulation of immune responses. The past year has seen considerable progress in the identification of intracellular molecules mediating CD40 signaling. Furthermore, it has been established that ligation of CD40 ligand (CD40L) delivers signals to the CD40L bearing cells themselves. Finally, the critical role of CD40-CD40L interactions in the development of various disease states has been fully appreciated. PMID- 9203419 TI - Role of phosphatases in lymphocyte activation. AB - Many lymphocyte signaling pathways are regulated by protein tyrosyl phosphorylation, which is controlled by protein tyrosine kinases (PTKs) and protein tyrosine phosphatases (PTPs). Substantial progress has been made in defining the functions of lymphocyte PTPs. Individual PTPs can enhance or diminish cell signaling levels. The transmembrane PTP CD45 is a key positive element in multiple lymphocyte signaling pathways in vivo. New insights into the function of individual CD45 isoforms have emerged. Anti-CD45 antibodies with potent immunosuppressant activity have been identified, suggesting that CD45 may be a propitious target for drug design. Progress has also been made in elucidating the function and targets of specific nontransmembrane PTPs, particularly those with Src homology 2 domains. PMID- 9203420 TI - Interactions of TCRs with MHC-peptide complexes: a quantitative basis for mechanistic models. AB - The activation of T lymphocytes is initiated by the binding of MHC-peptide complexes on antigen-presenting cells to MHC-restricted, peptide specific TCRs. Significant progress has recently been made in understanding the structure of the TCR and in the direct quantitative examination of the primary binding interactions between MHC-peptide complexes and the TCR. Attempts to develop quantitative models for the differential activation of T cells by MHC-peptide ligands that differ subtly in their structure have largely been based on either the affinity of the MHC-peptide complexes for the TCR in question or on the dissociation kinetics of the MHC-peptide complex from the T cell. PMID- 9203421 TI - The complexity of signaling pathways activated by the BCR. AB - Cross-linking of the B cell antigen receptor (BCR) leads to the activation of three types of intracellular protein tyrosine kinases. These tyrosine kinases then phosphorylate signaling components to activate a variety of signaling reactions, including phosphatidylinositol 4,5-bisphosphate hydrolysis, Ras activation, and phosphatidylinositol 3-kinase activation. Each of these signaling reactions, and also the signaling molecules Vav and HS1, appears to be important for at least some of the many types of B cell responses to antigen. The complexity of BCR signaling reactions may be required to allow the B cell to respond in a number of distinct ways to antigen (proliferation, survival, apoptosis, maturational arrest, etc.) depending on the maturation state of the B cell, the location in the body, the physical nature of the antigen, and the possible presence of the antigen in complex with antibody or complement components. PMID- 9203422 TI - Co-stimulation in T cell responses. AB - Antigen-specific T cell responses have primarily been considered in terms of activation signals delivered through the TCR and the co-stimulatory molecule CD28. In the past few years, studies have demonstrated the critical importance of inhibitory signals for regulating lymphocyte activation. CD28 and its homologue cytotoxic T lymphocyte antigen-4 (CTLA-4) share the same counter-receptors on antigen-presenting cells, but recent experiments have shown that CD28 and CTLA-4 have opposite effects on T cell activation. The mechanisms responsible for integrating these activation and inhibitory signals at the cellular and molecular levels are just beginning to be elucidated. PMID- 9203423 TI - Lymphocyte activation and effector functions. How signals are integrated in the immune system. PMID- 9203424 TI - Residential exposure to magnetic fields and acute lymphoblastic leukemia in children. AB - BACKGROUND: Previous studies found associations between childhood leukemia and surrogate indicators of exposure to magnetic fields (the power-line classification since known as "wire coding"), but not between childhood leukemia and measurements of 60-Hz residential magnetic fields. METHODS: We enrolled 638 children with acute lymphoblastic leukemia (ALL) who were under 15 years of age and were registered with the Children's Cancer Group and 620 controls in a study of residential exposure to magnetic fields generated by nearby power lines. In the subjects' current and former homes, data collectors measured magnetic fields for 24 hours in the child's bedroom and for 30 seconds in three or four other rooms and outside the front door. A computer algorithm assigned wire-code categories; based on the distance and configuration of nearby power lines, to the subjects' main residences (for 416 case patients and 416 controls) and to those where the family had lived during the mother's pregnancy with the subject (for 230 case patients and 230 controls). RESULTS: The risk of childhood ALL was not linked to summary time-weighted average residential magnetic-field levels, categorized according to a priori criteria. The odds ratio for ALL was 1.24 (95 percent confidence interval, 0.86 to 1.79) at exposures of 0.200 mu T or greater as compared with less than 0.065 mu T. The risk of ALL was not increased among children whose main residences were in the highest wire-code category (odds ratio as compared with the lowest category, 0.88; 95 percent confidence interval, 0.48 to 1.63). Furthermore, the risk was not significantly associated with either residential magnetic-field levels or the wire codes of the homes mothers resided in when pregnant with the subjects. CONCLUSIONS: Our results provide little evidence that living in homes characterized by high measured time-weighted average magnetic-field levels or by the highest wire-code category increases the risk of ALL in children. PMID- 9203425 TI - Use of inhaled corticosteroids and the risk of cataracts. AB - BACKGROUND: The use of systemic corticosteroids is a risk factor for the development of posterior subcapsular cataracts, but the association between inhaled corticosteroids and cataracts is uncertain. METHODS: We conducted a population-based, cross-sectional study of vision and common eye diseases in an urban area of the Blue Mountains, near Sydney, Australia. We recruited 3654 people 49 to 97 years of age; the participation rate was 82 percent. We collected information by questionnaire on potential risk factors for cataracts, including the current or prior use of inhaled corticosteroids (beclomethasone or budesonide). Photographs of the subjects' lenses were graded, without information on the subjects, to determine the presence and severity of cortical, nuclear, and posterior subcapsular cataracts. RESULTS: Three hundred seventy subjects reported using inhaled corticosteroids, 164 currently and 206 previously. Among these subjects, after adjustment for age and sex, there was a higher prevalence of nuclear cataracts (relative prevalence, 1.5; 95 percent confidence interval, 1.2 to 1.9) and posterior subcapsular cataracts (relative prevalence, 1.9; 95 percent confidence interval, 1.3 to 2.8) than among the subjects with no inhaled corticosteroid use, but the prevalence of cortical cataracts was not significantly higher (relative prevalence, 1.1; 95 percent confidence interval, 0.9 to 1.3). Higher cumulative lifetime doses of beclomethasone were associated with higher risks of posterior subcapsular cataracts (P for trend <0.001); the highest prevalence (27 percent) was found in subjects whose lifetime dose was over 2000 mg (relative prevalence, 5.5). Adjusting for the use of systemic corticosteroids and other potential confounders had little effect on the magnitude of the associations. The associations with posterior subcapsular cataracts, but not those with nuclear cataracts, were less marked when the analyses were restricted to subjects who had never used systemic corticosteroids. CONCLUSIONS: The use of inhaled corticosteroids is associated with the development of posterior subcapsular and nuclear cataracts. PMID- 9203427 TI - Fetal alloimmune thrombocytopenia. AB - BACKGROUND: Alloimmune thrombocytopenia is a serious fetal disorder resulting from platelet-antigen incompatibility between the mother and fetus. The diagnosis is usually made after the discovery of unexpected neonatal thrombocytopenia. Approximately 10 to 20 percent of affected fetuses have intracranial hemorrhages, one quarter to one half of which occur in utero. We studied the correlates of thrombocytopenia in affected fetuses. METHODS: We studied 107 fetuses with alloimmune thrombocytopenia at a mean (+/-SD) gestational age of 25+/-4 weeks, before their entry into one of three treatment protocols. The fetuses were initially evaluated because an older sibling had been given this diagnosis at birth. We compared the initial platelet counts in utero in these 107 fetuses with the platelet count at birth and the history of intracranial hemorrhage in the affected sibling. RESULTS: The initial platelet count was < or =20,000 per cubic millimeter in 53 of the 107 fetuses (50 percent), including 21 of 46 fetuses (46 percent) studied before 24 weeks of gestation. The 97 fetuses with PI(A1) incompatibility had more severe thrombocytopenia than the 10 fetuses with other antigen incompatibilities. Among seven fetuses with platelet counts of more than 80,000 per cubic millimeter that were not treated initially, the counts decreased by more than 10,000 per cubic millimeter per week. Although 41 fetuses had initial platelet counts that were lower than those measured at birth in an older affected sibling, only a history of antenatal intracranial hemorrhage in the sibling predicted greater severity of thrombocytopenia in the fetus. Only one treated fetus had an intracranial hemorrhage, and the thrombocytopenia resolved after birth in all cases. CONCLUSIONS: Fetal alloimmune thrombocytopenia occurs early in gestation, is severe, and is more severe in fetuses with an older affected sibling who had had an antenatal intracranial hemorrhage. PMID- 9203426 TI - Treatment of cryptococcal meningitis associated with the acquired immunodeficiency syndrome. National Institute of Allergy and Infectious Diseases Mycoses Study Group and AIDS Clinical Trials Group. AB - BACKGROUND: Treatment with low-dose amphotericin B (0.4 mg per kilogram of body weight per day) or oral azole therapy in patients with the acquired immunodeficiency syndrome (AIDS) and cryptococcal meningitis has been associated with high mortality and low rates of cerebrospinal fluid sterilization. METHODS: In a double-blind multicenter trial we randomly assigned patients with a first episode of AIDS-associated cryptococcal meningitis to treatment with higher-dose amphotericin B (0.7 mg per kilogram per day) with or without flucytosine (100 mg per kilogram per day) for two weeks (step one), followed by eight weeks of treatment with itraconazole (400 mg per day) or fluconazole (400 mg per day) (step two). Treatment was considered successful if cerebrospinal fluid cultures were negative at 2 and 10 weeks or if the patient was clinically stable at 2 weeks and asymptomatic at 10 weeks. RESULTS: At two weeks, the cerebrospinal fluid cultures were negative in 60 percent of the 202 patients receiving amphotericin B plus flucytosine and in 51 percent of the 179 receiving amphotericin B alone (P=0.06). Elevated intracranial pressure was associated with death in 13 of 14 patients during step one. The clinical outcome did not differ significantly between the two groups. Seventy-two percent of the 151 fluconazole recipients and 60 percent of the 155 itraconazole recipients had negative cultures at 10 weeks (95 percent confidence interval for the difference in percentages, -100 to 21). The proportion of patients who had clinical responses was similar with fluconazole (68 percent) and itraconazole (70 percent). Overall mortality was 5.5 percent in the first two weeks and 3.9 percent in the next eight weeks, with no significant difference between the groups. In a multivariate analysis, the addition of flucytosine during the initial two weeks and treatment with fluconazole for the next eight weeks were independently associated with cerebrospinal fluid sterilization. CONCLUSIONS: For the initial treatment of AIDS associated cryptococcal meningitis, the use of higher-dose amphotericin B plus flucytosine is associated with an increased rate of cerebrospinal fluid sterilization and decreased mortality at two weeks, as compared with regimens used in previous studies. Although consolidation therapy with fluconazole is associated with a higher rate of cerebrospinal fluid sterilization, itraconazole may be a suitable alternative for patients unable to take fluconazole. PMID- 9203428 TI - Simultaneous human granulocytic ehrlichiosis and Lyme borreliosis. PMID- 9203429 TI - Images in clinical medicine. Rheumatic mitral stenosis. PMID- 9203430 TI - Valvular heart disease. PMID- 9203431 TI - The new surrogates for board certification -- what should the standards be? PMID- 9203432 TI - Power lines, cancer, and fear. PMID- 9203433 TI - Cataracts and inhaled corticosteroids. PMID- 9203434 TI - The Kassebaum-Kennedy bill--the limits of incrementalism. PMID- 9203435 TI - Science: moving us in the right direction. PMID- 9203436 TI - Diabetic nephropathy: incidence, prevalence, and treatment. PMID- 9203438 TI - Growth patterns of large-for-gestational-age and appropriate-for-gestational-age infants of gestational diabetic mothers and control mothers at age 1 year. AB - OBJECTIVE: The purpose of this study was to explore the development of adiposity in macrosomic and normosomic infants of mothers with gestational diabetes mellitus (GDM) and control subjects between birth and age 1 year, and assess its relation to maternal prenatal factors and neonatal factors. RESEARCH DESIGN AND METHODS: This was a prospective observational study of 192 infants, including 47 large-for-gestational-age (LGA) infants of GDM mothers, 47 appropriate-for gestational-age (AGA) infants of GDM mothers, 55 LGA control infants, and 44 AGA control infants who were evaluated at birth and age 1 year. Maternal prenatal and pregnancy anthropometric measurements were recorded. Multiple infant anthropometric measurements, including skinfold thicknesses, were obtained at birth and age 1 year. Regression models were run to detect the independent effects of various maternal and infant factors on 1-year child adiposity, adjusting for their effects at birth. RESULTS: LGA infants of GDM mothers had a higher BMI, waist circumference, and abdominal skinfold at age 1 year compared with all other study groups. Among infants of GDM mothers, the mean 2-h postprandial glucose value for the second and third trimester correlated with waist circumference (r = 0.28, P < 0.04) and subscapular skinfold (r = 0.37, P < 0.007), and correlated marginally with 1-year sum of four skinfolds. Among infants of GDM mothers, a regression of 1-year sum of four skinfolds was significantly related to maternal prepregnancy weight after controlling for sum of skinfolds at birth. For control infants, the maternal glucose screen value was significantly associated with 1-year sum of skinfolds adjusted for the birth sum of skinfolds. CONCLUSIONS: We concluded that macrosomic infants of GDM mothers have unique patterns of adiposity that are present at birth and persist at age 1 year. Further, we concluded that maternal factors, including adiposity and intrauterine fuel environment, influence the presence and distribution of adiposity for both infants of GDM mothers and control infants. PMID- 9203437 TI - Stability of insulin lispro in insulin infusion systems. AB - OBJECTIVE: To test stability of insulin lispro in two insulin infusion systems over 48 h. RESEARCH DESIGN AND METHODS: We used reverse-phase and size-exclusion high-performance liquid chromatography (HPLC) to determine the purity, potency, and degree of polymerization of U100 insulin lispro (Humalog) after 24- and 48-h pump cycles conducted at 37 degrees C in five Disetronic H-TRON V100 and five MiniMed 504 pumps. Pumps were set to deliver a basal rate of 0.5 U/h and 6-U boluses at t = 0, 4, 8, 24, 24.5, 28.5, 32.5, and 48 h during each cycle. The effluent was collected into 1-ml vials, pooled at 24 or 48 h, and stored at 4 degrees C until assay. After each 48-h run period of insulin delivery, assays for potency, polymer, and purity were performed on the pooled samples from each individual cycle. m-cresol content and the pooled reservoir content were assayed in the 48-h pooled samples. RESULTS: Insulin lispro retained full HPLC potency (delta < or = 4%) at 48 h, with no degradation of insulin lispro to des amidoinsulin forms (24 or 48 h). No increase in pumped insulin polymer concentration was observed following 24 h of pump flow. Nonsignificant increases of < or =0.09% (Disetronic) and < or =0.15% (MiniMed) from initial concentrations of 0.18% (polymer divided by total insulin) were detected in three of five pump cycles at 48 h when compared with 37 degrees C paired controls. Nonsignificant decreases (<5 and 10%, Disetronic and MiniMed, respectively) of m-cresol content occurred in both systems following 48 h storage in each device, but sterility was not compromised by this decrease (initial m-cresol concentration, 3.15 mg/ml). Pump performance was without mechanical or electrical fault throughout the study Basal and bolus insulin delivery was evaluated three times daily and remained as expected. Occlusion of catheters by insulin precipitation did not occur, and no change in pH was observed following delivery. CONCLUSIONS: We conclude that insulin lispro is suitable for prolonged infusion in these two medical devices when syringes and catheters are replaced at 48-h intervals. PMID- 9203439 TI - Same eyes, different doctors: differences in primary care physician referrals for diabetic retinopathy screening. AB - OBJECTIVE: To analyze eye care specialist referral patterns for the diabetic patients of primary care physicians. RESEARCH DESIGN AND METHODS: In 1993, we conducted a census of primary care physicians to evaluate practice patterns relating to diabetes care in the state of Indiana. Using a logistic regression model and data from this census, we compared 1) physicians' odds of referring type II diabetic patients to an optometrist, as opposed to an ophthalmologist, with those of type I diabetic patients and 2) the referral odds ratios of type II to type I diabetic patients between metropolitan and nonmetropolitan counties. RESULTS: Overall, 10% of the physicians in our study most often refer some patients to an optometrist. Physicians are more likely to refer their type II diabetic patients to an optometrist, as opposed to an ophthalmologist, than they are to refer type I diabetic patients, both before and after adjustment for covariates. Physicians who practice in metropolitan counties are 1.55 times more likely to refer their type II diabetic patients than their type I diabetic patients to an optometrist. In nonmetropolitan counties, physicians are 2.5 times more likely to refer their type II diabetic patients to an optometrist. The difference between metropolitan and nonmetropolitan physicians is significant (P = 0.027). CONCLUSIONS: Some physicians mostly refer their diabetic patients to optometrists, instead of ophthalmologists, for eye examinations intended to discover early signs of diabetic eye disease. Type II diabetic patients are more likely to be referred to an optometrist, instead of an ophthalmologist, than are type I diabetic patients. In nonmetropolitan areas, the difference in referral patterns becomes even more marked. PMID- 9203440 TI - Diabetes nutrition and complications trial (DNCT): food intake and targets of diabetes treatment in a sample of Spanish people with diabetes. Diabetes and Nutrition Study Group of the Spanish Diabetes Association (GSEDNu). AB - OBJECTIVE: To know the nutritional pattern of people with diabetes in Spain and that pattern's relationship with targets of metabolic control. RESEARCH DESIGN AND METHODS: The Diabetes Nutrition and Complications Trial (DNCT) is a prospective multicenter study designed to determine the nutritional behavior of diabetic patients in Spain using 7-day food diaries. A total of 337 diabetic patients, 144 (70 men and 74 women) with type I and 193 (81 men and 112 women) with type II diabetes, satisfactorily completed the 7-day food diaries from May 1993 to December 1994. RESULTS: The median energy intake of the Spanish diabetic subjects was between 1,453 and 2,217 kcal/day (6.1-9.3 MJ/day), distributed as follows: 38-40% from carbohydrate, 19-23% from protein, and 36-41.5% from fat (11.1-13.9% saturated fatty acids, 4.6-5.8% polyunsaturated fatty acids, and 19.7 21.9% monounsaturated fatty acids). Of the subjects, 69% had a cholesterol level <5.6 mmol, 97% had an HDL cholesterol level >0.9 mmol, and 85% had a triglyceride level <1.7 mmol, while <36% had an HbA(1c) value >8%. CONCLUSIONS: People with diabetes in Spain have near-optimal serum lipid levels and maintain reasonably good blood glucose control and BMI, suggesting that diabetes management that includes the usual Spanish diet, which is low in carbohydrates and high in fat, especially monounsaturated fat, might be adequate. PMID- 9203441 TI - The unchanging incidence of hospitalization for diabetic nephropathy in a population-based cohort of IDDM patients in Finland. AB - OBJECTIVE: Finland has the highest documented incidence of childhood IDDM in the world, but the incidence of diabetic nephropathy in Finland is unknown. The aim of the present study was to determine the incidence of hospitalization for diabetic nephropathy in a population-based cohort of Finnish IDDM patients and to analyze the prognostic effect of sex, age at diagnosis, and calendar year of diagnosis of IDDM. RESEARCH DESIGN AND METHODS: We included all Finnish patients who had onset of IDDM before age 18 years, were diagnosed between January 1965 and December 1979 (n = 5,149), and were traced for hospitalizations between January 1970 and the end of December 1989 in the Hospital Discharge Register, using the unique personal identification code given to all Finnish citizens. The development of diabetic nephropathy was defined as the first hospitalization with a diagnosis of nephropathy (International Classification of Diseases-8th Revision [ICD-8] 250.04, or 9th Revision [ICD-9] 2503B/2503X). RESULTS: Among the 5,149 patients included, we identified 446 cases of diabetic nephropathy. The incidence of hospitalization for diabetic nephropathy was very low during the first 8 years of diabetes duration, and after that increased to a maximum of 1.6-2.0% per year. Female subjects developed nephropathy slightly earlier than male subjects, but the cumulative risk was independent of sex. Patients diagnosed at ages 5-14 years had the highest risk of hospitalization for diabetic nephropathy. We observed no effect of calendar year of diagnosis. CONCLUSIONS: We found a 20% cumulative incidence of hospitalization for diabetic nephropathy during a total 24 years of IDDM duration. This finding is compatible with the cumulative incidence of hospitalization for diabetic nephropathy found in other European populations. The incidence of hospitalization for diabetic nephropathy did not decrease during the 20-year observation period. PMID- 9203442 TI - The homeostasis model in the San Antonio Heart Study. AB - OBJECTIVE: Both insulin resistance and decreased insulin secretion have been shown to predict the development of NIDDM. However, methods to assess insulin sensitivity and secretion are complicated and expensive to apply in epidemiological studies. The homeostasis model assessment (HOMA) has been suggested as a method to assess insulin resistance and secretion from the fasting glucose and insulin concentrations. However, this method has not been extensively evaluated, particularly in different ethnic groups. RESEARCH DESIGN AND METHODS: We applied the HOMA model to cross-sectional analyses of the San Antonio Heart Study (n = 2,465). RESULTS: HOMA insulin resistance (IR) was very strongly correlated with fasting insulin (r = 0.98) and HOMA beta-cell function (beta cell) was moderately correlated with the 30-min increment in insulin concentration over the 30-min increment in glucose concentration (delta I30/delta G30) in an oral glucose tolerance test (OGTT) (r = 0.44). NIDDM was characterized by both high HOMA IR and low HOMA beta-cell function. In Mexican-Americans, HOMA IR in NIDDM subjects was 9.5 compared with 2.7 in normal glucose tolerance (NGT) subjects. In contrast, HOMA beta-cell function showed only small differences in Mexican-Americans (176 NIDDM; 257 NGT). However, the delta I30/delta G30 (pmol/mmol) showed much larger differences (75 NIDDM; 268 NGT). When modeled separately, impaired glucose tolerance (IGT) was characterized by high HOMA IR and high HOMA beta-cell function. However, when analyzed in the same regression model, high HOMA IR and low HOMA beta-cell function characterized subjects with IGT. These results were similar in both ethnic groups. Mexican-Americans had increased insulin resistance (as judged by both HOMA IR and fasting insulin) and insulin secretion (by HOMA beta-cell and delta I30/delta G30) relative to non Hispanic whites. CONCLUSIONS: We conclude that HOMA provides a useful model to assess insulin resistance and beta-cell function in epidemiological studies in which only fasting samples are available and that, further, it is critical to take into account the degree of insulin resistance in assessing insulin secretion by the HOMA model. PMID- 9203443 TI - The relationship of concentrations of insulin and proinsulin-like molecules with coronary heart disease prevalence and incidence. A study of two ethnic groups. AB - OBJECTIVE: To define the potential role of proinsulin-like molecules as risk factors for cardiovascular disease. RESEARCH DESIGN AND METHODS: Fasting concentrations of proinsulin, des-31,32-proinsulin, and insulin, and of insulin 2 h after a 75-g glucose load, were measured in 1,034 nondiabetic europid subjects and 257 south Asian subjects and related to prevalent coronary heart disease (Minnesota-coded electrocardiographic criteria or ischemic chest pain). In 137 south Asian subjects, the fasting concentrations were related to incident coronary heart disease over a 6.5-year follow-up. RESULTS: The standardized odds ratios for prevalent coronary heart disease were as follows: fasting insulin, 1.29 (1.11-1.49), P = 0.0006; 2-h insulin, 1.25 (1.08-1.45), P = 0.003; proinsulin, 1.23 (0.99-1.53), P = 0.058; and des-31,32-proinsulin, 1.32 (1.03 1.69), P = 0.026. The odds ratios were similar in the two ethnic groups. These relationships became insignificant when controlling for age, sex, and BMI. The standardized odds ratios for incident coronary heart disease were as follows: fasting insulin, 0.99 (0.63-1.55), P = 0.97; proinsulin, 1.13 (0.72-1.78), P = 0.59; and des-31,32-proinsulin, 1.00 (0.61-1.63), P = 1.00. CONCLUSIONS: We have found similar relationships between concentrations of proinsulin-like molecules and prevalent coronary heart disease, as are observed for insulin in these nondiabetic subjects, although these molecules comprise only approximately 10% of all insulin-like molecules. It appears biologically implausible that these relationships represent cause and effect. PMID- 9203444 TI - Transient impaired glucose tolerance in South African Indians does not carry a risk for progression to NIDDM. AB - OBJECTIVE: To evaluate the significance of transient impaired glucose tolerance (IGT) in terms of the risk of progression to NIDDM and the serum insulin response during oral glucose tolerance test (OGTT) in a prospective study on the natural history of IGT in South African Indians. RESEARCH DESIGN AND METHODS: This is a report on 87 subjects who formed part of a 4-year prospective study in 128 subjects classified with IGT at baseline (year 0) using World Health Organization criteria for glucose tolerance. Subjects were reexamined at years 1 and 4. At year 1, based on OGTT results, the subjects were divided into three groups: transient IGT (normal glucose tolerance [trIGT], n = 40), persistent IGT (pIGT, n = 47), and diabetes (n = 41). Analysis was performed on the 87 subjects who were classified as IGT at year 0, but who had not progressed to NIDDM by year 1 of the study At baseline (year 0), a modified OGTT was performed; between years 1 and 4, the OGTT included timed midtest samples for plasma glucose and serum insulin. Analysis of predictive factors for progression to diabetes or reversion to normal glucose tolerance was undertaken using year 0 as baseline. RESULTS: By year 4, 72 subjects (82.8%) completed the study Of the 32 subjects in the trIGT group, none (0%) had progressed to NIDDM, 11 (34.4%) had reverted to IGT (N-IGT), and 21 (65.6%) had persisted with normal glucose tolerance (N-N); of the 40 subjects in the pIGT group, 16 (40%) had progressed to NIDDM (IGT-D), 17 (42.5%) had persisted with IGT (IGT-IGT), and 7 (17.5%) had reverted to normal glucose tolerance (IGT-N). Significant predictive factors for reversion to normal glucose tolerance included absence of obesity (P = 0.0131, odds ratio [OR] 4.2, 95% CI 1.4-13.1) and 2-h plasma glucose level (P = 0.027, OR 2.4, 95% CI 1.11-5.13) at baseline (year 0). Intergroup (cross-sectional) analysis showed that the serum insulin response was higher in the pIGT than in the trIGT subgroup (fasting serum insulin: IGT-N vs. N-IGT and N-N, 16.9 +/- 1.9 vs. 6.8 +/- 2.1 and 6.1 +/- 2.4 microU/ml, respectively, P < 0.001; 2-h postload serum insulin: IGT-IGT vs. N IGT, 116.8 +/- 2.2 vs. 60.3 +/- 1.7 microU/ml, P < 0.05). By contrast, the insulinogenic index was higher in the trIGT subgroups both at year 1 (90-min: N-N vs. IGT-D, 48.9 +/- 3.9 vs. 14.1 +/- 2.5; P < 0.05) and at year 4 (N-N vs. remaining four subgroups, P < 0.01 at 60 min and 90 min). Intragroup (prospective) comparisons showed that in the N-IGT subgroup, the mean 2-h insulinogenic index was lower at year 4 than at year 1 (19.9 +/- 1.7 vs. 66.0 +/- 2.7; P < 0.05). CONCLUSIONS: In this 4-year prospective study in South African Indians, transient IGT carries no risk of progression to NIDDM. The significant predictive factors for reversion to normal glucose tolerance include lower baseline obesity prevalence and 2-h postload plasma glucose level. Moreover, in this group, beta-cell secretory function appeared to deteriorate with worsening of glucose tolerance. PMID- 9203445 TI - Low prevalence of antibodies to GAD65 in a 50- to 74-year-old general Dutch population. The Hoorn Study. AB - OBJECTIVE: To assess the prevalence of antibodies to GAD65 (GAD65-A) in relation to glucose tolerance disturbances and to blood glucose-lowering therapy in a general Dutch population. RESEARCH DESIGN AND METHODS: A population sample of 2,350 Dutch subjects, age 50-74 years, agreed to undergo an oral glucose tolerance test (OGTT). They were classified as having normal glucose tolerance, impaired glucose tolerance, newly detected diabetes, or known diabetes. GAD65-A levels were measured in serum by means of a standardized radioligand assay and subsequently were expressed as indexes. The prevalence rates were defined as the proportions of individuals of each category of glucose tolerance exceeding the value of the index at the 99th percentile of the entire study population. RESULTS: The prevalence rates and the 95% CIs of GAD65-A were 0.7% (0.4-1.2%) in cases of normal glucose tolerance, 2.4% (0.9-5.3%) in impaired glucose tolerance, 0% (0-3.3%) in newly detected diabetes, according to the World Health Organization (WHO) criteria, and 3.5% (0.7-10.0%) in known diabetes. A total of 2 out of 3 subjects with GAD65-A indexes above the 99th percentile and 10 out of 18 subjects with GAD65-A indexes above the 85th percentile received insulin therapy for their diabetes, which showed an association between GAD65-A and insulin therapy CONCLUSIONS: Low prevalence rates of latent autoimmunity to GAD were found in 50- to 74-year-old Dutch subjects with normal and abnormal glucose tolerance, and GAD65-A was associated with insulin use in known diabetic subjects. PMID- 9203446 TI - Elevated glycosylated albumin in NIDDM is a function of recent everyday environmental stress. AB - OBJECTIVE: To evaluate the association of recent daily environmental stress (daily hassles) with glycemia in NIDDM. RESEARCH DESIGN AND METHODS: Fifty-five NIDDM patients reported the number and intensity of daily hassles occurring during the past week and concurrently underwent glycemic assessment. RESULTS: Hassles were generally unassociated with demographic variables, illness duration, treatment regimen, and the presence of complications. Multiple regression analysis indicated that hassles (in both frequency and intensity) were positively associated with recent glycemia (glycosylated albumin [GA]), even after statistically controlling for long-term glycemia (glycosylated hemoglobin [HbA(1c)]). The subtypes of hassles having the most potent relationships with GA were work and family/friend-related stressors. CONCLUSIONS: The frequency and perceived impact of everyday minor stress have proximal positive associations with glycemia that do not necessarily reflect chronic hyperglycemia. Stress arising from work and family/friend sources may be particularly relevant. PMID- 9203447 TI - Open-flow microperfusion of subcutaneous adipose tissue for on-line continuous ex vivo measurement of glucose concentration. AB - OBJECTIVE: To evaluate a novel technique for on-line continuous glucose measurement in subcutaneous adipose tissue, and to investigate its accuracy for detection of hypoglycemia. RESEARCH DESIGN AND METHODS: The method combined an open-flow microperfusion of subcutaneous adipose tissue using a double lumen catheter and an extracorporeal sensor cell. An isotonic ion-free solution was perfused through the inner lumen of the catheter, equilibrated with the subcutaneous tissue fluid, and sampled through the outer lumen. The recovery was continuously monitored as the ratio between the measured sampled fluid conductivity and the subcutaneous tissue fluid conductivity (assumed to have a constant value of 1.28 S/m at 25 degrees C). Glucose concentration was calculated on-line from the measured glucose in the sampled fluid and the measured recovery in healthy volunteers during hyperglycemic glucose loads (n = 8), hypoglycemic hyperinsulinemic clamp (n = 6), and a 24-h monitoring period (n = 7). RESULTS: Subcutaneous glucose concentrations in the fasting state were 94% of the plasma glucose concentrations in arterialized venous samples. According to the error grid analysis, 96.9% of the on-line measured subcutaneous glucose concentrations during hyperglycemia and 96.3% during hypoglycemia were in accurate or acceptable zones. The mean differences between the measured subcutaneous glucose and the actual plasma glucose concentration were -0.06-3.3 mmol/l (hyperglycemia), and 0.6-1.1 mmol/l (hypoglycemia). CONCLUSIONS: By combining open-flow microperfusion, glucose sensor, and conductivity measurement, glucose concentration in the subcutaneous adipose tissue can be monitored on-line, extracorporeally, and continuously without any in vivo calibration, and gives accurate measurements during hyper- and hypoglycemia. PMID- 9203448 TI - Dexfenfluramine in obese Chinese NIDDM patients. A placebo-controlled investigation of the effects on body weight, glycemic control, and cardiovascular risk factors. AB - OBJECTIVE: To investigate the safety, efficacy, and metabolic effects of dexfenfluramine in obese Chinese NIDDM patients. RESEARCH DESIGN AND METHODS: Thirty-two patients, mean (+/- SD) body weight 76.2 +/- 8.5 kg with corresponding BMI 31.1 +/- 2.1 kg/m2, were randomized into a two-phase study, after a 2-week single-blind run-in period on placebo. Phase 1 was a randomized 3-month double blind placebo-controlled trial during which either dexfenfluramine or placebo was added to the existing treatment regimens of diet plus or minus sulfonylureas without metformin. Phase 2 was a further 3-month single-blind trial during which the placebo group was given dexfenfluramine without patients' knowledge of changing to active medication, while the active group continued with dexfenfluramine. Body weight, glycemic control, blood pressure, lipids, and quality of life were assessed before and at 3 and 6 months after randomization. A total of 27 patients were also followed for an additional period of 6-12 months (215 +/- 53 days) after dexfenfluramine treatment was withdrawn. RESULTS: During the run-in period, both groups were comparable for all parameters measured. At 3 months, mean changes in BMI were -1.2 +/- 1.0 kg/m2 (dexfenfluramine) vs. -0.1 +/ 0.5 kg/m2 (placebo) (P < 0.001). The mean changes in fasting plasma glucose were -1.14 +/- 0.99 vs. 0.51 +/- 1.34 mmol/l (dexfenfluramine vs. placebo, P = 0.004). HbA(1c) also significantly improved in the dexfenfluramine group (-0.80 +/- 0.53 vs. 0.25 +/- 0.64%, P < 0.001). During the 3-month single-blind dexfenfluramine treatment in the ex-placebo group, there were similar improvements in body weight and glycemic indexes. After cessation of dexfenfluramine therapy at 6 months, significant increases in body weight and glycemic indexes, almost back to the baseline, were observed for both groups. CONCLUSIONS: Dexfenfluramine aids weight loss and improves glycemic control in obese Chinese NIDDM patients over a 3- to 6 month period. These effects are emphasized after withdrawal of treatment and further support the longer-term use of dexfenfluramine for chronic complicated obesity. PMID- 9203449 TI - A neural network model for predicting pancreas transplant graft outcome. AB - OBJECTIVE: To compare the results of a neural network versus a logistic regression model for predicting early (0-3 months) pancreas transplant graft survival or loss. RESEARCH DESIGN AND METHODS: This study was a cross-sectional, secondary analysis of demographic and clinical data from 117 simultaneous pancreas-kidney (SPK), 35 pancreas-after-kidney (PAK), and 8 pancreas-transplant alone (PTA) patients (n = 160). The majority of patients were men (57%) and were white (90.1%), with a mean age of 39 +/- 8.09 years. Of the patients, 23 (14.4%) experienced early graft loss, which included any loss owing to technical or immunological causes, and death with a functional graft. Data were analyzed with a logistic regression model for multivariate analysis and a backpropagation neural network (BPNN) model. RESULTS: A total of 12 predictor variables were chosen from literature and transplant surgeon recommendations. A logistic model with all predictor variables included correctly classified 93.53% of cases. Model sensitivity was 35.71%; specificity was 100% (pseudo-R2 0.24). Of the predictors, history of alcohol abuse (odds ratio [OR] 32.39; 95% CI 1.67-626.89), having a PAK or PTA (OR 13.6; 95% CI 2.20-84.01), and use of a nonlocal organ procurement center (OPO) (OR 4.51; 95% CI 0.78-25.96) were most closely associated with early graft loss. The BPNN model with the same 12 predictor variables correctly predicted 92.50% of cases (R2 0.71). Model sensitivity was 68%; specificity was 96%. Of the predictors, the three variables most closely associated with graft outcome in this model were recipient/donor weight difference >50 lb, having a PAK or PTA, and use of a nonlocal OPO. CONCLUSIONS: First, the BPNN model correctly predicted 92.5% of graft outcomes versus the logistic model (93.53%). Second, the BPNN model rendered more accurate predictions (>0.70 = loss; <0.30 = survival) versus the logistic model (>0.50 = loss; <0.50 = survival). Third, the BPNN model was more sensitive (68%) than the logistic model (35.71%) to graft failures and demonstrated an almost threefold increase in explained variance (R2 = 0.71 vs. 0.24). These results suggest that the BPNN model is a more powerful tool for predicting early pancreas graft loss than traditional multivariate statistical models. PMID- 9203450 TI - Erythromycin derivative improves gastric emptying and insulin requirement in diabetic patients with gastroparesis. AB - OBJECTIVE: To evaluate the effect of the erythromycin derivative EM523L on gastric emptying and postprandial insulin requirement in insulin-dependent diabetic patients with severe gastroparesis. RESEARCH DESIGN AND METHODS: In six IDDM patients with severe gastroparesis (two men and four women, mean age 44.5 years [range 36-53]), the insulin infusion pattern during feedback control with an artificial endocrine pancreas device (Biostator) after intake of a test meal, the retention rate of residual isotope ([99m]Tc-labelled Sn-colloid) in the stomach, and the time-concentration curve of plasma acetaminophen as the marker for liquid emptying were studied with EM523L or a control placebo RESULTS: Time courses of insulin infusion rates peaked within 120 min after intake of the test meal in the EM523L phase, whereas no apparent peak rates were observed in the control phase. The total amount of insulin required in the first 90 min postprandial was significantly greater in the EM523L phase than in the control phase. EM523L significantly decreased the residual isotope ratio in the stomach at > or =50 min postprandial and increased the plasma acetaminophen concentrations at 30-120 min postprandial, compared with respective values in the control phase. CONCLUSIONS: Preliminary results obtained from a small number of patients suggest that EM523L or erythromycin analogs, which have agonistic activity to motilin receptors as well as no antibacterial effect, may be useful to accelerate gastric emptying and improve insulin requirement patterns, thereby establishing more stable glycemic control. PMID- 9203451 TI - Diabetes associated with a novel 3264 mitochondrial tRNA(Leu)(UUR) mutation. AB - OBJECTIVE: To present a novel mitochondrial DNA mutation in a diabetic family RESEARCH DESIGN AND METHODS: The proband was a 64-year-old man. In the family, diabetes was maternally inherited. He had diabetes, cerebellar ataxia, cervical lipoma, hearing loss, olfactory dysfunction, ophthalmoplegia, and facial nerve bilateral palsy. On examination, early insulin secretion was blunted, and the M value on glucose clamp test was low. In muscle, ragged red fibers were not found. T-to-C mutation at position 3264 was detected in the proband (0.5% mutant DNAs in leukocyte and 30% in muscle), but was not detected in 201 normal individuals. RESULTS: Heteroplasmy of mutation, maternal inheritance of diabetes, and symptoms related to mitochondrial dysfunction suggest the pathogenecity of this 3264 mutation. As for diabetes etiology, both impaired insulin secretion and decreased insulin sensitivity seem to be important. In phenotypic characteristics, the combination of cerebellar ataxia and lipoma is a symptom sometimes found in myoclonic epilepsy and ragged red fibers (MERRFs). Ophthamoplegia is a symptom of chronic progressive external ophthalmoplegia (CPEO). These suggest that our proband had phenotypic overlap with MERRF and CPEO. Conversely, facial nerve bilateral palsy is a rare finding. The pictures that focused on his cranial nerves were thus unique, suggesting the heterogeneity of mitochondrial DNA (mtDNA)-related diabetes. CONCLUSIONS: A novel 3264 mitochondrial DNA mutation in diabetes gives new insight to the etiology of mitochondrial diabetes. Its pathogenecity supports the belief that the tRNA(Leu)(UUR) gene is an etiological hot spot of mitochondrial diseases. PMID- 9203452 TI - Blood glucose concentration influences postprandial fullness in IDDM. AB - OBJECTIVE: Upper gastrointestinal (GI) symptoms and delayed gastric emptying both occur frequently in patients with long-standing IDDM, but the relationship between them is relatively weak. Recent studies in normal subjects have indicated that blood glucose concentration may increase the perception of sensations arising from the upper GI tract. The purpose of this study was to examine the relationships among postprandial fullness, the rate of gastric emptying, and blood glucose concentration in IDDM patients. RESEARCH DESIGN AND METHODS: We studied measurements of gastric emptying, blood glucose concentrations, cardiovascular autonomic nerve function, upper GI symptoms, and postprandial hunger and fullness in 40 IDDM patients (16 men, 24 women). ages 19-63 years. Gastric emptying of solids and liquids was measured scintigraphically, upper GI symptoms were measured by questionnaire immediately before ingestion of the test meal, and fullness and hunger were measured by visual analog scales every 15 min. Blood glucose concentrations were measured at -5, 30, 60, 90, and 120 min. RESULTS: Solid gastric emptying was delayed in 58% of the patients, and both solid and liquid gastric emptying were slower (P < 0.05) in women than in men. The score for upper GI symptoms was not significantly related to gastric emptying. In contrast, postprandial fullness, but not hunger, was related to the amount of solid (r = 0.36, P < 0.05) but not liquid in the stomach. Both before (r = 0.39, P < 0.05) and after (r = 0.47, P < 0.01) the meal, fullness was related to blood glucose concentration. Postprandial fullness was also related to autonomic nerve dysfunction (r = 0.39, P < 0.05). Multiple regression analysis confirmed that blood glucose concentration, the rate of solid gastric emptying, and autonomic nerve dysfunction were independent determinants of postprandial fullness, together accounting for 47% of the variance. CONCLUSIONS: These observations demonstrated that, in IDDM, postprandial fullness is influenced by blood glucose concentration, the rate of solid gastric emptying, and autonomic nerve function. PMID- 9203454 TI - Long-term hyperglycemia is related to peripheral nerve changes at a diabetes duration of 4 years. The Wisconsin Diabetes Registry. AB - OBJECTIVE: To examine longitudinal hyperglycemia and peripheral nerve responses in a population-based incident cohort. RESEARCH DESIGN AND METHODS: A sample from an incident cohort of young people was comprehensively followed from diagnosis of IDDM. Participants were invited to submit blood samples three times per year for central testing of GHb. During their 4th year of diabetes, nerve conduction studies were performed on the median sensory and motor, peroneal motor, and sural sensory nerves. Relationships between mean GHb and nerve latencies, velocities, and amplitudes were explored. RESULTS: GHb was positively related to all nerve latencies and negatively related to all nerve velocities. The relationships between mean GHb and nerve conduction latencies and velocities differed by sex for the peroneal nerve latency (beta = 0.17 male subjects, beta = -0.01 female subjects; P < 0.001). Pubertal participants had lower velocities and longer latencies than prepubertal participants (beta = 0.37; P = 0.05 peroneal latency), after adjustment for GHb, height, and extremity temperature. Sensory and motor nerve amplitudes were related to GHb, and these relationships did not differ by sex. CONCLUSIONS: Our study indicates that sustained hyperglycemia is related to functional changes, at the minimum, in peripheral sensory and motor nerve conduction at a diabetes duration of 4 years. Our findings are consistent with a dying-back neuropathy, and there is some suggestion that chronic hyperglycemia may be more detrimental to nerves in male subjects than in female subjects. PMID- 9203453 TI - Incidence of blindness in relation to diabetes. A population-based study. AB - OBJECTIVE: A reduction of diabetes-related blindness was declared a primary objective for Europe (St. Vincent Declaration). We collected data about incidence rates of blindness in the diabetic population compared with the nondiabetic population. Up to now, such data are scarce-even worldwide. RESEARCH DESIGN AND METHODS: A complete list of newly registered blindness allowance recipients was drawn up in the district of Wurttemberg-Hohenzollern, Germany, between 1990 and 1993. From these data, we estimated age-specific and standardized incidence rates of blindness in the entire, the diabetic, and the nondiabetic population, as well as relative and attributable risks due to diabetes. RESULTS: There were 2,714 people meeting the inclusion criteria; 1,823 (67.2%) were female and 781 (28.8%) had diabetes. In 318 subjects, diabetes was likely to be the only cause of blindness; in 192 subjects, it was one of several contributory causes. Age of women was 73.9 +/- 19.4 years (mean +/- SD) and of men 63.3 +/- 25.5 years. Results standardized to the (West) German population are as follows: incidence rates (per 100,000 person-years): total population: 13.5; diabetic population: 60.6; nondiabetic population: 11.6; relative risk: 5.2; attributable risk among exposed: 0.81; and population attributable risk: 0.14. The relative risks decreased considerably with increasing age. When the study is repeated to monitor the St. Vincent targets, a reduction in the incidence rate of blindness in the diabetic population by 17% will be detected with 95% power. CONCLUSIONS: Great relative and attributable risks, especially in younger age-groups, indicate the need for increased attention to preventive measures for microvascular complications. PMID- 9203455 TI - Clinical characteristics of Japanese men with glucokinase gene beta-cell promoter variant. AB - OBJECTIVE: To study the association between a variant at the position of -30 of beta-cell-specific promoter of the glucokinase gene and glucose tolerance in the Japanese general population and to assess the clinical characteristics of subjects with the variant. RESEARCH DESIGN AND METHODS: The genotype of 657 Japanese men aged 51.0 +/- 8.8 years (mean +/- SD) was analyzed by an allele specific assay using polymerase chain reaction-restriction fragment length polymorphism. RESULTS: The variant allele frequency was 0.188 in subjects with normal glucose tolerance, 0.211 in subjects with impaired glucose tolerance, and 0.176 in diabetic subjects. In subjects with fasting plasma glucose levels <140 mg/dl, homozygous subjects for the promoter variant had significantly higher plasma glucose levels 60 min after oral glucose administration when compared with subjects without the variant allele. A cross-sectional analysis showed age related elevation of basal glucose levels only in subjects without the promoter variant. Individuals heterozygous for the variant had significantly lower levels of HDL cholesterol than normal subjects. HDL cholesterol values were lower in homozygous people than in normal and heterozygous subjects, although the differences were not statistically significant. CONCLUSIONS: The beta-cell promoter variant in homozygous state was associated with impaired glucose tolerance, but not with diabetes, and low HDL cholesterol levels in Japanese men. It is unlikely that the glucose intolerance associated with the promoter variant is progressive with age. PMID- 9203456 TI - Risk factors for diabetic peripheral sensory neuropathy. Results of the Seattle Prospective Diabetic Foot Study. AB - OBJECTIVE: To identify risk factors for diabetic lower-extremity peripheral sensory neuropathy prospectively in a cohort of U.S. veterans with diabetes. RESEARCH DESIGN AND METHODS: General medicine clinic outpatients with diabetes were followed prospectively for the development of insensitivity to the 5.07 monofilament on the foot. RESULTS: Of 775 subjects, 388 (50%) had neuropathy at baseline. Of the 387 subjects without neuropathy at baseline, 288 were followed up, and of these, 58 (20%) developed neuropathy. Multivariate logistic regression modeling of prevalent neuropathy controlling for sex and race revealed independent and significant associations with age, duration of diabetes, glycohemoglobin level, height, history of lower-extremity ulceration, callus, and edema; an independent and inverse correlation was noted with ankle-arm index. Risk factors for incident neuropathy in multivariate logistic regression included age, baseline glycohemoglobin level, height, history of ulcer, and CAGE screening instrument alcohol score; current smoking and albumin level were inversely associated with risk. CONCLUSIONS: Poorer glycemic control increases the risk of neuropathy and is amenable to intervention. Height and age directly increase risk of neuropathy and may help identify patients at risk. A proportion of neuropathy in diabetic veterans is probably due to or worsened by alcohol ingestion. Neuropathy was less common in current smokers than subjects not currently smoking. PMID- 9203457 TI - Immune response to glycated and oxidized LDL in IDDM patients with and without renal disease. AB - OBJECTIVE: To study autoantibodies to oxidized and glycated LDL in IDDM patients with and without diabetic nephropathy and in nephropathy-related macroangiopathy RESEARCH DESIGN AND METHODS: The study included 101 IDDM patients with a long duration of diabetes and 54 healthy subjects. Patients were divided into two groups according to their median urinary albumin excretion rate (AER); the normoalbuminuric group had AER <20 microg/min and the albuminuric group >200 microg/min. The groups were matched for age and BMI, and the two diabetic groups were matched for duration of diabetes and glycemic control. Antibodies against oxidized LDL (using malondialdehyde-modified LDL as the antigen) and against glycated LDL were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: The mean antibody levels against glycated LDL were higher in IDDM patients (0.305 +/- 0.399) than in healthy subjects (0.166 +/- 0.22 optical density [OD]; P = 0.019), but levels did not differ significantly between normoalbuminuric and albuminuric IDDM patients (0.258 +/- 0.354 vs. 0.388 +/- 0.459, respectively). Among the three groups, antibody levels to oxidized LDL did not differ. IDDM patients showed an inverse correlation between antibodies to oxidized LDL and HbA1 (r = -0.211, P = 0.04). The antibody levels to glycated and oxidized LDL did not differ among albuminuric IDDM patients with or without clinical macroangiopathy. CONCLUSIONS: Antibodies to glycated and oxidized LDL do not seem to associate with diabetic nephropathy or nephropathy-related macroangiopathy. PMID- 9203458 TI - Insulin antibody response to a short course of human insulin therapy in women with gestational diabetes. AB - OBJECTIVE: To assess the insulin antibody (IA) response to human insulin (HI) therapy in women with gestational diabetes. RESEARCH DESIGN AND METHODS: IAs were measured by a competitive radiobinding assay in 50 women with gestational diabetes before and during treatment with HI and after delivery. At delivery, 15 maternal-cord blood sample pairs were analyzed for IA. As a reference, we searched for IA in 25 new-onset type I diabetic patients, before and at 3, 6, and 12 months after insulin therapy. RESULTS: Insulin autoantibodies (IAAs) were detected in 1 of 50 women with gestational diabetes and 4 of 16 type I diabetic patients (P < 0.05). At the end of pregnancy after 9.3 +/- 6.8 weeks on insulin therapy, 22 of 50 (44%) women with gestational diabetes became IA+ and 4 additional women were found to be positive 2 months postpartum. After 3 months on insulin, type I diabetic patients showed a higher rate of IA positivity (92%, P < 0.001). IA titers at the end of pregnancy were associated with the cumulative insulin dose (r = 0.29, P < 0.05). Postpartum, IA disappeared slowly in most IA+ women, but two women still showed IA 2 years after delivery Titers in cord blood were strongly related to those in maternal blood (r = 0.74, P < 0.01). The rate of adverse fetal outcome did not differ in IA and IA- mothers (27 vs. 40%, NS). CONCLUSIONS: HI is immunogenic, and a short course of HI therapy induces IA in approximately 50% of women with gestational diabetes and 92% of type I diabetic patients. In women with gestational diabetes, insulin dose is slightly associated with IA titers. These IAs apparently cross the placenta. Fetal outcome does not differ according to the maternal IA status, and IAs disappear gradually after delivery but may remain positive for 2 years after delivery. PMID- 9203459 TI - Physical activity efficacy and effectiveness among older adults and minorities. AB - The objective of this study was to consider efficacy and effectiveness of physical activity for the prevention and management of NIDDM among minorities and older adults of the U.S. Relevant population trends and projections are discussed, followed by a review of the efficacy of physical activity based on theoretical, prospective cohort, and intervention studies. With few empirical studies available, the assessment of effectiveness is largely theoretical and focuses on potentially important issues for future studies among older adults and minorities. Efficacy studies have shown that moderate-intensity physical activity is associated with a one- to two-thirds lower incidence of NIDDM over 4-14 years and 15-20% lower glycosylated hemoglobin over 3-4 months among people with NIDDM. With physical inactivity prevalence at 60-70%, much work remains to be done to improve physical activity effectiveness. In the most vulnerable populations, physician referral and community involvement structured around stage of change and self-efficacy theories are suggested as the most promising approaches to promoting physical activity adoption and maintenance. Effectiveness or demonstration studies that test and build on stage of change, self-efficacy, and other concepts of physical activity promotion and outcomes would likely prove to be highly valuable investments for public health. PMID- 9203460 TI - Report of the Expert Committee on the Diagnosis and Classification of Diabetes Mellitus. PMID- 9203461 TI - The 32nd Annual Meeting of the European Association for the Study of Diabetes. Macrovascular disease. PMID- 9203462 TI - Acanthosis nigricans and diabetic ketoacidosis. PMID- 9203463 TI - Down's syndrome, IDDM, and hypothyroidism. PMID- 9203464 TI - Does severity of neuropathy influence consent to invasive investigative procedures in clinical trials? Determinants for sural nerve biopsy. PMID- 9203465 TI - Evaluation of blood glucose meters during hypoglycemia. PMID- 9203466 TI - Accuracy of blood glucose meters during hypoglycemia. PMID- 9203467 TI - Hyperbaric oxygen therapy for diabetic foot ulcers. PMID- 9203469 TI - Choreoathetosis and diabetes. PMID- 9203468 TI - Self-monitoring of blood glucose compared with continuous tissue glucose measurement. PMID- 9203470 TI - Potentially lethal ileus associated with acarbose treatment for NIDDM. PMID- 9203472 TI - Reported rates, incentives, and effectiveness of major vaccinations in 501 attendees at two diabetes clinics. PMID- 9203471 TI - Spurious glycohemoglobin values associated with hydroxyurea treatment. PMID- 9203473 TI - Bloomgarden's February editorial on managed care. PMID- 9203474 TI - Early postprandial hypoglycemia following administration of lispro insulin. PMID- 9203475 TI - Glibenclamide and liver disease. PMID- 9203476 TI - Minimally invasive surgery: competent and continent? PMID- 9203477 TI - The contribution of urethrocystoscopy to evaluation of lower urinary tract dysfunction in women. AB - The aim of this study was to determine whether the evaluation of lower urinary dysfunction with urodynamics and urethrocystoscopy provides unique information that is missed by urodynamics alone. Eighty-four women underwent multichannel urodynamics and urethrocystoscopy. Retrospective analysis included evaluation of the relationships between lower urinary tract lesions and risk factors using chi2 and Fisher's exact tests. Urethrocystoscopic findings changed the diagnosis and management in 6 patients. New urethrocystoscopic findings included papillary transitional-cell carcinoma, cystitis glandularis, an intravesical suture and a urethral diverticulum. Clinical parameters were not predictive of these findings. Urethrocystoscopic findings also contributed to the final diagnosis in 10 patients with intrinsic sphincter deficiency. Considered alone, maximum urethral closure pressure < or =20 cmH2O had a sensitivity of only 20% and a positive predictive value of 40% for this diagnosis. Urodynamics without urethrocystoscopy would have missed important diagnoses in 19% of women. Urethrocystoscopy and urodynamics complement one another, and both have a role in the evaluation of women with lower urinary tract dysfunction. PMID- 9203478 TI - Intrasubject variability of the pressure-transmission ratio in patients with genuine stress incontinence. AB - The aim of the study was to determine the intrasubject variability of the pressure-transmission ratio (PTR) with various cough intensities in subjects with genuine stress incontinence. Thirty-six patients with genuine stress incontinence underwent multichannel urodynamics and had a series of pressure-transmission ratios (PTRs) determined with the urethral transducer placed at the point of the maximal closure pressure. Patients were asked to cough with increasing intensities and three to four different cough-induced PTRs were recorded for each subject. The data were analysed using regression analysis, repeated measures analysis of variance and comparison of variance. The PTRs showed a high degree of variability within subjects. The mean within subject standard deviation was 18.5%. The effect of parity, maximal urethral closure pressure and age were insignificant on the variability. Cough intensities of greater than 90 cmH2O have a lesser degree of variability. The mean PTR across all cough intensities was fairly constant in the 82%-87% range. It was concluded that the PTR in an individual has a high degree of variability independent of cough intensity, and cannot be relied upon as a diagnostic measure in subjects with genuine stress incontinence. However, the PTR for the population as a whole was consistent across all cough intensities. PMID- 9203479 TI - Dynamic assessment of pelvic floor function in women using the intravaginal device test. AB - The aim of the study was to assess pelvic floor function and dysfunction using intravaginal devices (IVD test). One hundred and eighty-five patients were evaluated, 65 (35.1%) in the control group without genital prolapse and 120 (64.9%) in the study group, with prolapse. Anatomic changes were evaluated on a scale described by Halban, and functional classification based on palpation of the muscles of the pelvic floor during contraction. Additionally, weighted vaginal devices were used to assess pelvic floor function. Statistic analysis was performed with the Spearman-Pearson correlation coefficient, the chi2 test and the response/operator characteristic curve. There was an acceptable correlation between the IVD test and the functional classification of 0.75. Using this classification, the IVD test showed 86.58% sensitivity, 75.72% specificity, and had a positive predictive value of 73.95% and a negative predictive value of 87.64%. Significant differences between pelvic floor muscle activity in those patients with and without genital prolapse were observed (chi2 = 58.28, P=<0.005). It was concluded that pelvic floor assessment can be done through the evaluation of active muscle strength or pelvic floor integrity using the functional classification and the IVD test. PMID- 9203480 TI - Magnetic resonance imaging of the pelvic floor in the postpartum patient. AB - Magnetic resonance imaging (MRI) was used to assess anatomical changes in the pelvic floor after childbirth. Six women underwent serial MRI examination within 30 hours and at 1 week, 2 weeks, 6 weeks and 6 months after delivery; 8 additional women were studied only within 30 hours of delivery. T-1 and T-2 weighted images of the pelvis in the transverse and sagittal planes with a 1.5-T MR imager were obtained. In the sagittal section we assessed the urethrovesical angle, urethral length, distance from the symphysis to the proximal and distal vagina, vaginal length, width and length of the sphincters, and the presence of sphincter defects. Axial sections were assessed for sphincter defects for the distance between the symphysis and midurethra, vagina and rectum. Only one parameter (distance between symphysis and distal vagina) changed significantly over time, without a clear trend in direction. Interobserver variation was reasonable (<15%) except for anal canal length, urethral length and distance between symphysis and anus. There were no significant correlations between birthweight and MRI parameters. There was a non-significant association (P = 0.09) between the sole combined sphincter defect and rectal injury, but not with episiotomy or parity. We concluded that it is feasible to determine multiple measurements on MR images to evaluate structures of the pelvic floor. PMID- 9203481 TI - Direct effect of amezinium on rabbit urethra: effect of estrogen and progesterone treatment. AB - The effect of amezinium, a new anti-hypotensive agent, and hormonal treatment on the female rabbit urethra was investigated. Cumulative dose responses were obtained for amezinium and norepinephrine on strips of muscle from the urethras of ovariectomized female rabbits by means of the tissue-bath system. Amezinium enhanced the response to electrical field stimulation and showed a direct contractile response on the urethra. These responses were only about 20% of the maximum norepinephrine response. The contractile response to amezinium was completely blocked by prazosin. When rabbits were pretreated with estrogen, with or without progesterone, for 4 weeks, the response to amezinium increased to 40% of the maximum norepinephrine response. Although amezinium enhances muscle contractile responses to electrical stimulation, this effect is strong when amezinium is used alone; concurrent estrogen treatment improves the effects of amezinium. PMID- 9203482 TI - Sling transection of urethra: a rare complication. AB - Common postoperative complications associated with suburethral sling procedures include voiding disorders and urinary retention, de novo development of detrusor instability, sling graft rejection and, rarely, erosion of the graft into the urethra. The authors present a case of a late postoperative complication of polytetrafluoroethylene graft erosion and partial transection of the urethra, with resultant acute urinary retention. A 50-year-old patient presented with acute urethral outflow obstruction due to sling graft erosion into the urethra nearly 2 years after she underwent a curative sling procedure for recurrent genuine stress incontinence. After relieving the acute urinary retention by inserting a suprapubic catheter under ultrasound guidance, the sling graft was accessed and removed. The urethral defect was repaired successfully. At follow-up 5 months later, the patient was continent subjectively and by urodynamic criteria, with no voiding abnormalities. Although erosion of the sling graft into the urethra and transection of this structure is a rare complication after a sling procedure, it should be considered in the patient who experiences progressive voiding difficulties, has transvaginal urinary leakage, and/or cannot be catheterized transurethrally. Expedient relief of the urinary retention and outflow obstruction is necessary, as well as careful surgical reconstruction of the urethra. To minimize the development of this complication we recommend plication of paraurethral connective tissue in the midline beneath the sling graft, and placement of minimal tension on the sling. PMID- 9203483 TI - Periurethral collagen implant: ultrasound assessment and prediction of outcome. AB - The objective of this study was to identify sonographic parameters that could predict successful outcome in women after periurethral collagen implant. Thirty one women with a diagnosis of stress urinary incontinence with intrinsic sphincteric deficiency underwent one periurethral collagen implant between January and December 1994. Three months after the procedure ultrasound evaluation was performed using a 5 MHz probe placed at the vaginal introitus. Subjective assessment and cough stress test were used to measure outcomes. Twenty-five women were available for evaluation 1 year after the procedure. A successful outcome was found in 18 of the 25 women subjectively (72%) and in 16 objectively (64%). A distance of the collagen from the bladder neck of less than 7 mm was found to be associated with a positive outcome. This threshold was found to have a sensitivity of 83.3%, specificity of 85.7%, a positive predictive value of 93.7% and a negative predictive value of 66.6%. PMID- 9203484 TI - Biofeedback and physiotherapy versus physiotherapy alone in the treatment of genuine stress urinary incontinence. AB - Biofeedback is a method of pelvic floor rehabilitation using a surface electrode inserted into the vagina and a catheter in the rectum. Forty women with genuine urinary stress incontinence were randomized to compare the efficacy of physiotherapy and physiotherapy in combination with biofeedback. The effect of the treatment was determined by a standardized pad-weighing test. Long-term status was determined using a questionnaire after 2-3 years. Thirty-four women completed the treatment. The study showed a statistically significant better improvement in the biofeedback group. The long-term effect in the biofeedback group seemed better and the patients were more motivated for training afterwards. PMID- 9203485 TI - Evisceration after enterocele repair: a rare complication of vaginal surgery. AB - Evisceration is a life-threatening surgical emergency which must be promptly treated. Evisceration following vaginal enterocele operation is so rare that no incidence rate can be established. A review of the literature revealed only 71 cases. The reported women were in general postmenopausal, with only 15% less than 50 years of age. In 18% of cases vaginal rupture occurred while straining at stool. Four patients are known to have died due to evisceration. Of the reported repair operations, 57% were performed transabdominably, 28% transvaginally, and 15% via a combined abdominovaginal route. The authors present 1 more case treated transvaginally, with a review of the literature. PMID- 9203486 TI - The first century of urogynecology and reconstructive pelvic surgery: where do we go from here? Presidential Address, 21st Annual Clinical Meeting, International Urogynecological Association, Vienna, Austria, 2-5 September 1996. PMID- 9203488 TI - Transesophageal echocardiography with the use of a four-millimeter probe. AB - Transesophageal echocardiography with the use of pediatric probes is nowadays commonly performed. However, in small children, insertion of a probe with a diameter of 7 mm may be traumatic or even impossible. We therefore tested a 17 element, 4 mm transverse plane probe in 136 pediatric patients, mainly in the operation room, catheterization laboratory, or the intensive care unit, and in three healthy adult volunteers. This probe was easy to insert, particularly during emergency situations, did not cause any complication in any patient, and provided satisfactory information despite the low number of elements. The use of a 4 mm transesophageal probe can improve the management of neonates with congenital heart disease in the operating room or the neonatal intensive care unit. PMID- 9203487 TI - Toroidal geometry: novel three-dimensional intracardiac imaging with a phased array transducer. AB - Recent advances in small, linear-array transducers have opened new avenues for three-dimensional image acquisition from an intracardiac approach. The purpose of this study was to introduce a novel method of image acquisition using toroidal geometry, explore its fidelity of reproduction of three-dimensional cardiac anatomy, and determine whether a whole-heart scan is achievable. Acquisition was accomplished through 360-degree incremental rotation of a rigid endoscope with a side-mounted ultrasound transducer. The procedure was first tested with the use of a gelatin model to define far-field slice resolution with 1.8-degree rotational increments. Comparison of three-dimensional scans of cardiac specimens with corresponding photographs confirmed that toroidal geometry can provide a high-quality display of structures from all sides. We conclude that whole-heart three-dimensional scanning from within the cardiac chambers is possible with toroidal geometry. The quality of depicted anatomy depends on transducer location within the heart, distance from the transducer, density of slices, and image resolution. The potential of intracardiac three-dimensional ultrasound imaging includes detailed spatial evaluation of cardiac morphology, determination of appropriate placement of investigative or therapeutic devices (catheters, closure devices, etc.), and assessment of cardiac function. PMID- 9203489 TI - Echocardiographic imaging of a basket catheter for mapping and ablation of ventricular tachycardia in pigs. AB - Our objective was to assess the feasibility and efficacy of the recently described left ventricular simultaneous deployment of a new multi-electrode mapping catheter and a standard radio-frequency ablation catheter in pigs, with echocardiography monitoring and fluoroscopy guidance. Introduction and deployment of both catheters in five healthy anesthetized pigs were guided on-line by fluoroscopy and monitored with transthoracic echocardiography. Heart rate and femoral blood pressure were also continuously monitored. Both catheters were deployed for up to 5 hours. Three animals underwent three to five radio-frequency energy applications. Left ventricular dimensions obtained from long axis two dimensional echocardiography imaging before and after basket-catheter deployment in the left ventricular cavity, were 3.9 +/- 0.3 versus 3.7 +/- 0.6 cm at end diastole and 2.8 +/- 1.1 versus 2.6 +/- 0.8 cm at end-systole, respectively (mean +/- standard error of the mean, p > 0.05). Shortening fraction measured from long axis two-dimensional echocardiography images before and after catheter deployment was 28% +/- 10% versus 25% +/- 5%, respectively (mean +/- standard error of the mean, p > 0.05). Additional findings included the following: (1) good conformation of the multi-electrode mapping catheter to the left ventricular dimensions during diastole; (2) absence of catheter-induced aortic and/or mitral insufficiency, as well as left ventricular outflow tract obstruction; (3) absence of damage to mitral and aortic valves or to the left ventricular wall. Postmortem examination and hemodynamic measurements confirmed these findings and showed only minor subendocardial hemorrhages; (4) radio-frequency energy application produced intracavitary bubbles, which were demonstrable echocardiographically, enabling identification of the gross anatomic location of ablation sites. Echocardiography during simultaneous deployment of multi-electrode mapping catheter and radio frequency ablation catheters enables estimation of mechanical interaction with the left ventricle and detects interaction with myocardial/valvular function. During radio-frequency energy application, bubble production may identify gross anatomic location of ablation. PMID- 9203490 TI - Comparison of integrated backscatter values obtained with acoustic densitometry with values derived from spectral analysis of digitized signals from a clinical imaging system. AB - Time-domain-based integrated backscatter values obtained with the use of acoustic densitometry (AD) were compared with values determined from a spectral-based analysis of the radio-frequency (RF) signals with a modified Hewlett-Packard Sonos 1500 imaging system. Integrated backscatter images of five specimens of bovine tendon were acquired in the AD acquisition mode, and the corresponding signals related to the backscattered RF were digitized for each angle of insonification as the specimens were rotated in 10-degree increments. The integrated backscatter images were analyzed with the AD analysis package, and the corresponding values determined from the RF power spectra were obtained from the digitized ultrasonic signals. Good agreement was found between the two methods over the entire range of measured values. The mean anisotropy in the measured integrated backscatter (mean +/- standard error) was found to be 27 +/- 2 dB for time-domain-based analysis and 25 +/- 2 dB for RF spectral-based analysis. PMID- 9203491 TI - Left ventricular Doppler filling pattern in dilated cardiomyopathy: relation to hemodynamics and left atrial function. AB - This study attempted to examine the relation of left ventricular filling patterns to hemodynamic status and left atrial function in dilated cardiomyopathy. Transesophageal echocardiography and cardiac catheterization were performed in 41 patients with dilated cardiomyopathy (six with an ischemic origin). Transmitral, left atrial appendage, and pulmonary venous flow velocities were recorded with the pulsed Doppler method. Left atrial systolic function was assessed by the peak velocity of the left atrial appendage emptying wave and pulmonary venous flow reversal during atrial systole. Patients were classified into three groups according to their left ventricular filling patterns. Group 1 showed an abnormal relaxation pattern (E wave/A wave ratio <1, n = 17), group 2 had a normal or pseudonormal pattern (1 < or = E/A < 2, n = 11), and group 3 had a restrictive pattern (E/A > or = 2, n = 13). No differences were found among the groups with regard to age, gender, heart rate, and M-mode echocardiographic indices of left ventricular function. Compared with patients in group 1, those in groups 2 and 3 had more symptoms (New York Heart Association functional class III or IV) and had higher left ventricular filling pressures. The sensitivity of an E/A ratio > or = 1 for predicting a pulmonary capillary wedge pressure > or = 15 mm Hg was 75% and the specificity was 94%. Despite a similar increase of filling pressures, group 3 patients had a lower left atrial appendage emptying velocity, pulmonary venous flow reversal velocity, and mitral A velocity than did group 2 patients. The sensitivity and specificity of an E/A ratio > or = 22 for detecting left atrial dysfunction (left atrial appendage emptying velocity < or = 40 cm/sec) was 85% and 86%, respectively. In conclusion, among patients with dilated cardiomyopathy, those who had a restrictive or pseudonormal filling pattern were in a higher functional class and had higher filling pressures. Further studies are needed to determine the therapeutic and prognostic significance of left atrial dysfunction, which was common in patients with a restrictive pattern. PMID- 9203492 TI - Doppler echocardiography in cardiac transplant patients: allograft rejection and its relationship to diastolic function. AB - Right ventricular endomyocardial biopsy has been the traditional gold standard for determining cardiac transplant rejection. Although endomyocardial biopsy has proved useful in guiding rejection therapy, this procedure is not without risk. The objective of the present study was to determine whether a noninvasive method for assessing cardiac diastolic function would be of value in predicting biopsy scores. Doppler echocardiographic indices of left ventricular function were compared with biopsy scores in 43 studies from 23 patients (age 50 +/- 8 years). The average time from transplant to echocardiographic study was 1.5 years. Standard clinical indices of diastolic function failed to predict biopsy results. The A-Ar interval, evaluated in 36 studies, was found to significantly decrease (p < 0.003) with increasing biopsy scores. Preliminary results suggest that this echocardiographic parameter may prove useful in predicting biopsy results. PMID- 9203493 TI - Clinical and echocardiographic correlates of mitral E-wave transmission inside the left ventricle: potential insights into left ventricular diastolic function. AB - The mitral inflow wave is initially directed to the left ventricular apex and then turns around facing the left ventricular outflow tract. The E and A waves are transmitted to the left ventricular outflow tract where they are registered as Er and Ar waves, respectively. We hypothesized that the E-wave transit time to the left ventricular outflow tract recorded as the E-Er interval may depend on left ventricular early diastolic performance such as relaxation. This hypothesis was tested in clinical settings known to have abnormal left ventricular relaxation. Mitral E and left ventricular outflow tract Er waves were recorded with pulsed wave Doppler technique in 63 subjects: 25 healthy subjects, 18 patients with secondary left ventricular hypertrophy, and 20 patients with hypertrophic cardiomyopathy. The E-Er interval was measured from the onset of E wave to the onset of Er wave timed to the R wave of the electrocardiogram. The E Er interval ranged from 45 to 300 msec: 96 +/- 28 msec in the controls, 127 +/- 46 msec in patients with left ventricular hypertrophy (p = 0.0091 versus controls), and 179 +/- 57 msec in patients with hypertrophic cardiomyopathy (p < 0.0001 versus controls). It correlated with left ventricular free wall thickness (r = 0.42, p = 0.0006), thickness of the ventricular septum (r = 0.43, p = 0.0004), left ventricular end-diastolic diameter (r = -0.38, p = 0.0022), left ventricular end-systolic diameter (r = -0.55, p < 0.0001), left ventricular isovolumic relaxation time (r = 0.39, p = 0.0063), RR interval (r = 0.28, p = 0.045), mitral E/A velocity ratio (r = -0.33, p = 0.010), and E-wave deceleration time (r = 0.38, p < 0.0044) but not with age. Multivariate analysis with all the previously mentioned variables and the group the patient belonged to as the dichotomous variable showed that the grouping variable was the sole independent determinant of the E-Er interval (multiple r = 0.74). The E-Er interval is an easily measurable Doppler parameter which is increased in left ventricular hypertrophy and hypertrophic cardiomyopathy. It is related to left ventricular wall thickness, left ventricular isovolumic relaxation time, mitral E/A velocity ratio, and E-wave deceleration time and may provide useful insight into left ventricular early diastolic performance-possibly the relaxation process. PMID- 9203494 TI - Transesophageal echocardiographic Doppler study of the pulmonary venous flow pattern in severe mitral stenosis with variable degrees of mitral regurgitation. AB - The transesophageal echocardiographic data of 62 patients with severe, rheumatic mitral stenosis, which was either isolated or associated with different degrees of mitral regurgitation were reviewed to study and compare their pulmonary venous flow patterns. Peak systolic and peak diastolic flow velocities and their respective time intervals were measured, and the presence or absence of systolic flow reversal (SFR) was noted. The venous flow velocities and time integrals were all below normal and the ratio between the systolic and diastolic velocities were all blunted. Systolic flow reversal was observed in some patients with severe mitral stenosis with or without mitral regurgitation, and was highly correlated with the presence of atrial fibrillation. Among patients with mitral regurgitation and in atrial fibrillation, flow reversal timing was shorter in patients with significant mitral regurgitation than in patients with mild or no mitral regurgitation. PMID- 9203495 TI - Subaortic septal bulge simulates hypertrophic cardiomyopathy by angulation of the septum with age, independent of focal hypertrophy. An echocardiographic study. AB - Focal hypertrophy of the basal anterior septum occurs not infrequently in elderly patients and is considered by some to be a significant form of hypertrophic cardiomyopathy; others consider it to be an unimportant anatomic variant associated with an angulated septum, called a septal bulge (SB). We analyzed 94 cases of SB collected prospectively and compared them with 88 patients with extensive hypertrophic cardiomyopathy (HCM), 20 patients with hypertrophic cardiomyopathy limited to the entire septum (ASH), and 20 age-matched controls. The SB cases were also divided into three groups, with marked, moderate, or no basal septal hypertrophy associated with the occurrence of an SB. All groups of SB patients had increased fractional shortening compared with controls (0.48 +/- 0.07 versus controls 0.40 +/- 0.07), comparable with HCM (0.48 +/- 0.12), and increased left ventricular outflow tract velocity both at rest and especially after amyl nitrite inhalation (3.42 +/- 1.35 versus 1.55 +/- 0.60 m/sec [controls]). Other features of HCM were not present: normal wall thickness except for the basal septal hypertrophy, no anterior malposition in SB patients, no age independent reversal of ratio of early to late mitral inflow velocity (E/A), and no decrease in end-diastolic dimension. It is concluded that outflow tract narrowing by an angulated septum is the primary mechanism responsible for the increased outflow tract velocity, rather than the hypertrophic septum. The resultant increase in convective acceleration simulates the dynamics of hypertrophic cardiomyopathy. The focal hypertrophy may be secondary and contributory to the enhanced ventricular dynamics, but it does not appear to be a primary cardiomyopathy. PMID- 9203496 TI - Tetralogy of Fallot with absent pulmonary valve: echocardiographic morphometric features of the right-sided structures and their relationship to presentation and outcome. AB - Respiratory symptoms in tetralogy of Fallot with absent pulmonary valve are believed to be due to bronchial compression secondary to dilated pulmonary arteries; however, not all patients are born compromised. Echocardiographic morphometry of the right-sided structures was investigated to determine the possible relationship between anatomy and clinical presentation. Twenty-five patients were identified, and 15 had preoperative echocardiograms. Patients were divided into two groups: those with respiratory distress (group I, n = 9) and those without (group II, n = 6). No difference was noted in branch pulmonary artery diameters between groups; however, the pulmonary valve/ aortic valve ratio, reflecting the dimension of the narrowest pathway from the right ventricle, was larger in group I (0.74 +/- 0.15 versus 0.60 +/- 0.07, p < 0.05). Pulmonary valve diameter correlated with main and right pulmonary artery diameters. We conclude that patients with tetralogy of Fallot with absent pulmonary valve and respiratory compromise have a greater pulmonary valve/aortic valve ratio but do not have greater dilatation of proximal branch pulmonary arteries. This suggests that the pathophysiology is not due solely to compression of the bronchi but is also related to the blood flow dynamics in the pulmonary vessels. PMID- 9203497 TI - The ability of vegetation size on echocardiography to predict clinical complications: a meta-analysis. AB - To clarify whether echocardiographic detection of a vegetation 10 mm or larger in size in patients with left-sided infective endocarditis poses an increased risk for complications, we performed a meta-analysis of English-language publications identified by a computerized search of the key words infective endocarditis and echocardiography. A pooled odds ratio was calculated by using the Robins, Greenland, and Breslow estimate of variance. The pooled odds ratio for increased risk of systemic embolization in the presence of a vegetation >10 mm (10 studies, 738 patients) was 2.80 (95% confidence interval [CI] 1.95 to 4.02; p < 0.01). The odds ratio of requiring valve-replacement surgery (seven studies, 549 patients) was 2.95 (95% CI 1.90 to 4.58; p < 0.01). The odds ratio of death (six studies, 476 patients) was 1.55 (95% CI 0.92 to 2.60; p = 0.10). Thus this analysis supports the hypothesis that echocardiographically detected left-sided vegetations >10 mm pose a significantly increased risk of (1) systemic embolization and (2) a need for valve-replacement surgery than cases where either no or smaller vegetations are detected. PMID- 9203498 TI - Systemic embolization by a thrombus in a apparently normal aorta detected with transesophageal echocardiography. PMID- 9203499 TI - Diagnosis of a persistent coronary fistula after ventricular septal defect patch closure. AB - Penetrating chest trauma can result in multiple clinical syndromes depending on the structures involved. Tamponade, valvular regurgitation, ventricular septal defect (VSD), conduction system abnormalities, and coronary lacerations have been reported. We report a case of right ventricular free wall laceration, VSD, and coronary artery fistula involving a septal perforator. PMID- 9203500 TI - Acute myocardial infarction associated with dobutamine stress echocardiography. AB - Acute myocardial infarction as a complication of dobutamine stress echocardiography (DSE) is described in two patients during or shortly after undergoing the procedure. Both clinical events resulted in characteristic elevations in cardiac enzymes and the development of new electrocardiographic Q waves in the inferior leads. Subsequent coronary angiography was performed in both cases; one patient required two-vessel coronary artery bypass grafting to his first obtuse marginal and posterior descending arteries, and the other underwent successful angioplasty of an occluded proximal right coronary artery. Only two cases of DSE-associated myocardial infarction have been reported previously in the literature. PMID- 9203501 TI - Cardiac cavernous hemangioma mimicking pericardial cyst: atypical echocardiographic appearance of a rare cardiac tumor. AB - Cavernous hemangioma is a rare tumor with infrequent cardiac involvement. Preoperative or antemortem diagnosis may be difficult. Several prior case reports have described echocardiographic findings of cavernous hemangioma. We report here a 50-year-old white female patient with this tumor. Transesophageal echocardiography detected a mass with an echocardiographic appearance not previously described for cavernous hemangioma. The tumor appeared as a large echolucent unilocular cystic mass, leading to an erroneous preoperative diagnosis of pericardial cyst. This previously unreported finding should be recognized by echocardiographers in the evaluation of cardiac masses. PMID- 9203502 TI - Left ventricular pseudoaneurysm with left atrium tamponade: a rare postinfarction complication. AB - A 65-year-old man was readmitted to the hospital with deep-vein thrombophlebitis 3 weeks after suffering a small infarction. Two days after the patient was readmitted severe symptoms of cardiac failure developed. Transthoracic echocardiographic examination showed an unusual echo-free space adjacent to the lateral wall of the left atrium. No further diagnostic information could be obtained from precordial examinations. Single-plane transesophageal echocardiography demonstrated the presence of a ruptured aneurysm in the posterolateral wall of the left ventricle and a pseudoaneurysm, which was severely compressing the left atrial cavity. On color-flow Doppler examination a tear was detected in the wall of the left ventricular aneurysm close to the posterior atrioventricular sulcus. Pericardial adhesions because of coronary artery bypass grafting performed 10 years before may have contributed to the unusual location of this left ventricular pseudoaneurysm, complicated with left atrium tamponade after myocardial infarction. PMID- 9203504 TI - Enhanced proliferation and migration and altered cytoskeletal proteins in early passage smooth muscle cells from young and old rat aortic explants. AB - Smooth muscle cell (SMC) proliferation, migration, and cytoskeletal protein expression were studied in cultured cells obtained from the aortic explants of young (6-month) and old (30-month) Fischer 344XNB rats. Second-passage SMC were cultured on coverslips, and cytoskeletal fibers were examined by immunofluorescence microscopy using antibodies specific for smooth muscle myosin, alpha-smooth muscle actin, vimentin, desmin, and tubulin. The cytoskeletal fiber density was quantified as fluorescence intensity by confocal microscopy. The proliferation of SMC was analyzed from the growth curve of cells grown in culture from 0 to 14 days, and a Boyden chamber assay was used to quantify the SMC migration rate. The diameter of fresh SMC digested enzymatically from old rat aortae was 52.4% larger than that of the cells from young animals (20.0 +/- 3 microm vs 13.1 +/- 2 microm, P < 0.05). In SMC cultured from old animals, the intensities of smooth muscle myosin, alpha-smooth muscle actin, and vimentin decreased by 59.6, 41.2, and 54.8%, respectively; desmin and tubulin increased by 46.1 and 65.1% (all P < 0.001). Compared to SMC isolated from young rat aortae, the number of SMC cultured (second passage) from the old rat aorta was increased by 48.4, 27.2, and 26.9%, respectively, at Days 3, 7, and 14 in culture (P < 0.05, P < 0.01, and P < 0.001). The migration rate of SMC cultured from old rats was 59.3% higher than that of the cells obtained from young rats. These data show that alterations of the SMC cytoskeleton occur concomitantly with changes in SMC proliferation and migration rate during aging, suggesting that the age-associated changes in cytoskeletal proteins may play a role in remodeling of the aortic wall during aging. PMID- 9203503 TI - Arterial desaturation and orthodeoxia after atrial septal defect repair: demonstration of the mechanism by transesophageal and contrast echocardiography. AB - A young woman without symptoms underwent repair of an ostium secundum atrial septal defect, and exertional dyspnea developed postoperatively. This was found to be due to arterial oxygen desaturation which was exaggerated in the upright position and with exercise. Contrast echocardiography confirmed a right-to-left shunt at the atrial level that was shown only with femoral venous contrast injection and not with upper extremity venous injection. Transesophageal echocardiography and subsequent surgical exploration found that the Eustachian valve had been mistaken for the inferior rim of the defect and sutured to the upper rim of the defect. This created a channel through which blood from the inferior vena cava could be partially deferred to the left atrium. PMID- 9203505 TI - Quantitative ultrastructural evaluation of satellite cells in soleus muscle from rats kept in hypokinesia. AB - The aim of the present study is a morphometric, ultrastructural evaluation of satellite cells (SC) derived from the soleus muscle (SOL) of rats exposed to conditions of hypokinesia for a period of 7 or 21 days. Qualitative and quantitative analysis of 320 electron micrographs of SC from each group was carried out. After 7 and 21 days of immobilization, profiles of the SC, in contrast to the control group, had a lower mean surface area, had a cylindrical shape, and exhibited more folded membrane. Analysis of electron micrographs of SC showed that after immobilization, a lower number of SC contained profiles of mitochondria, rough endoplasmic reticulum (RER), and Golgi. Volume fractions of RER were twofold lower after 7 days of hypokinesia and fivefold lower after 21 days compared with the control group. The SC of SOL of rats subjected to hypokinesia differed from the control group by a markedly decreased number of ribosomes and RER profiles. After 21 days of immobilization the ultrastructural characteristics of SC were typical for cells in an inactive state showing various degrees of degeneration. The results of our study presented here permit the conclusion that 21 days of hypokinesia induce a depression of SC activity, whereas subtle changes in SC appear only after 7 days of immobilization. PMID- 9203506 TI - Ciliary neurotrophic factor immunoreactivity in rat intramuscular nerve during reinnervation through a silicone tube after severing of the rat sciatic nerve. AB - The immunoreactivity of ciliary neurotrophic factor (CNTF) and S100 was studied in the degenerating and regenerating intramuscular nerves after the sciatic nerve was severed. The sciatic nerves of male Wistar rats were transected at the midpoint of the thigh, and silicone tubing was used to obtain effective reinnervation. The strong immunoreactivity of CNTF and S100 was observed in the Schwann cell cytoplasm of intramuscular nerves (IMN) and at the neuromuscular junction (NMJ) on the control sections. The CNTF immunoreactivity gradually became weak and indistinct in the Schwann cell cytoplasm after the operation. However, it was recognized again in the IMN at 4 weeks after the operation. On the other hand, the S100 immunoreactivity was continuously observed except at the NMJ through the denervating and reinnervating period. At 12 weeks after the operation, the strong immunoreactivity of both CNTF and S100 was observed again. These findings suggest that the amount of CNTF protein decreased in Schwann cells of the IMN and NMJ during the denervating period and increased during the reinnervating period in proportion to the number of remyelinated Schwann cells after severing of the sciatic nerve. They also suggest that CNTF was more highly correlated than the S100 protein with the reinnervation activity of Schwann cells. PMID- 9203507 TI - Quantitation of angiogenesis in healing anastomoses of the rat colon. AB - An accurate and reliable quantitation of angiogenesis is an essential requirement for a detailed study of new blood vessel growth during healing of colon anastomoses. In the present study we applied a computer-based digital image processing system for quantitative analysis of the anastomotic vascularization and perfusion. Rats underwent colonic resection (1 cm) followed by construction of an end-to-end anastomosis. The animals were killed 3 or 7 days after operation. The vascularization and perfusion were analysed in cyrostat sections using an anti-collagen type IV antibody and the fluorescent marker bisbenzimide H 33342, respectively. If compared with 3-day-old anastomoses, a significant increase in the median vascular and perfusion areas was found 7 days after operation. Within the same anastomosis, the vessel density and perfusion were lower in the central region than in the peripheral region of the wound. The present computerized digital image analysis system is a reliable technique that is suited for quantitative measurements of microvascular changes occurring in colon anastomoses during the first postoperative week. PMID- 9203508 TI - Quantitation of vascular endothelial growth factor mRNA levels in human breast tumors and metastatic lymph nodes. AB - In situ hybridization analysis provides a means to qualitatively study the heterogeneity of primary tumors and metastases based on the types of genes transcribed. In this study, we have tested some parameters for quantitative analysis of in situ hybridizations with paraffin-embedded human breast tumors and measured mRNA levels for the angiogenic protein, vascular endothelial growth factor (VEGF). VEGF mRNAs were highly tumor specific, with the highest levels near necrotic regions within the tissues (0.1 to 2.7 dpm/mm2). Normal cells within the tissue sections did not have detectable levels of VEGF mRNA. For comparison, tumor levels of c-myc (4 to 46 dpm/mm2) and glyceraldehyde-3 phosphate dehydrogenase mRNAs (48 to 214 dpm/mm2) were measured. The mRNAs for both of these genes were more broadly expressed across the tissue sections. The hybridization pattern for VEGF mRNAs was consistent with hypoxia-induced VEGF mRNA steady-state levels and supports the hypothesis that oxidative stress regulates VEGF expression in breast tumors. PMID- 9203509 TI - Evidence for prothrombin production and thrombin expression by phorbol ester treated THP-1 cells. AB - With the addition of fibrinogen, fibrin clots form in serum-free culture medium recovered from phorbol ester-treated THP-1 cells. We attribute this coagulant activity to thrombin generated as a consequence of cell stimulation because the coagulant activity exists in serum-free culture medium from treated cells only, and it is inhibited by hirudin. The thrombin does not derive from a prothrombin/thrombin contaminant since no detectable prothrombin/thrombin preexists in either the serum-free culture medium or the fibrinogen preparations used for our experiments. We hypothesized that the thrombin is synthesized by the cells themselves. In support of this hypothesis, we found that prothrombin mRNA is expressed in THP-1 cells following their treatment with phorbol ester. Accompanying expression of this mRNA, prothrombin antigen becomes detectable in lysates of PMA-treated THP-1 cells, and thrombin antigen and activity become detectable in both lysates and culture medium of treated cells. These results are consistent with the notion that certain cells of myelomonocytic lineage are capable of synthesizing proteins relevant to coagulation. PMID- 9203510 TI - Cardiac contractility: modulation of myofibrillar calcium sensitivity by beta adrenergic stimulation. AB - Under conditions of beta-adrenergic receptor stimulation, cardiac performance is enhanced. cAMP-dependent phosphorylation of proteins located in the sarcolemma, in the membrane of the sarcoplasmic reticulum (SR), and in the myofibrils of the cardiomyocytes, mediates the effects of catecholamines on the heart. Altered Ca2+ handling leads to increased levels of intracellular free Ca2+. This is mainly responsible for the enhanced contractility of the myocardium that can be observed following beta-adrenergic receptor stimulation. Phosphorylation of the thin filament regulatory protein troponin I (TnI), on the other hand, decreases the Ca2+ sensitivity of the myofilaments, which means that the Ca2+ concentration necessary for the development of half-maximal force is increased. Cardiac TnI has a 26-33 amino acid N-terminal extension that is not present in fast and slow skeletal muscle TnI isoforms. Within this segment, two adjacent serine residues can be phosphorylated by a cAMP-dependent protein kinase. Replacement of endogenous TnI by different mutants obtained using site-directed mutagenesis of one or both of the serine residues has shown that only the bis-phosphorylated form decreases the Ca2+ sensitivity. This Ca2+ desensitizing effect, together with an increased rate of Ca2+ uptake into the SR due to phosphorylation of the SR membrane protein phospholamban, is responsible for the relaxation-enhancing effect (lusitropic action) of catecholamines. The latter is an important determinant of coronary perfusion and rapid diastolic filling of the ventricles, and is also a prerequisite for the elevation of heart rate that accompanies beta adrenergic receptor stimulation. PMID- 9203511 TI - The use of DNA markers in the pre-clinical diagnosis of familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP), an autosomal dominant inherited disease, confers a high risk of colon cancer. For presymptomatic diagnosis of FAP, we performed linkage studies in three unrelated Israeli families with FAP, using seven polymorphic systems around or at the APC locus on chromosome 5q. These systems are constituted of three DNA probes, recognizing four restriction fragment length polymorphism: C11p11, YN5.48 and pi227; three cytosine-adenine repeat markers: D5S318, D5S346 and MBC; and one intragenic polymorphism: APC SspI. A meiotic recombination event was detected, apparently between the FAP gene and probe pi227. Based on the different analysis systems, we determined the haplotype at the APC locus in 11 at-risk individuals of the three families, six of whom were found to carry the disease-linked allele. Additionally, we identified a new FAP patient, in whom sigmoidoscopy showed the presence of adenomatous polyps throughout the colon. PMID- 9203512 TI - Effects of oleic acid lung injury and positive end-expiratory pressure on central hemodynamics and regional blood flow. AB - The objective of our study was to investigate the influence on central and regional circulation of the application of positive end-expiratory pressure (PEEP=P) in a canine model of low hydrostatic pulmonary edema. Eight mongrel dogs with oleic acid-induced pulmonary edema were artificially ventilated with and without PEEP. Regional blood flow was determined using radioactive microspheres directly injected into the left ventricle. Regional blood flow to the brain was maintained under all experimental conditions, while the blood flow to the gastric fundal mucosa and to the pancreas significantly decreased following PEEP, oleic acid injection (OA) and with PEEP and oleic acid combination (P+OA). The renal blood flow decreased only during the P+OA phase. We conclude that the observed decrease in blood flow to the gastrointestinal mucosa and renal circulation in this acute low hydrostatic pressure pulmonary edema may correlate with the increased incidence of gastrointestinal and renal complications that accompany critically ill patients. PMID- 9203513 TI - Hepatitis C virus genotypes in patients with persistent infection--a preliminary report. AB - Hepatitis C virus (HCV) has nucleotide sequence diversity distributed throughout the viral genome, with variants showing even less than 70% homology. There is some evidence that sequence variation of HCV genotypes partly determines the course of infection and response to treatment with interferon. We studied the sera of 29 Israeli HCV patients, all suffering from chronic liver disease, and 34 patients with renal failure necessitating hemodialysis. HCV genotypes were detected using a reverse hybridization assay (LiPA), after reverse transcription polymerase chain reaction, using primers spanning the 5' UTR of the HCV genome. In this preliminary report the predominant HCV type detected was type 1, found in 65% of the chronic hepatitis patients and in 88% of the hemodialysis patients. Subtype 1b was the most prevalent and was detected in >40% of the chronic hepatitis patients and in >70%of the dialysis patients. Other types detected were 2a and 3, and in only two patients was type 4 found. More than 50% of patients with type 1 (1a or 1b) among patients with chronic hepatitis had received blood transfusion in the past, but only 16.6% of patients bearing subtype 2a HCV had such a history. Our preliminary evaluation revealed that patients bearing subtype 1b seemed to have a better response to interferon treatment, as compared with patients infected with subtypes 1a, 2a, who displayed a low response rate. PMID- 9203514 TI - Evaluation of domiciliary long-term oxygen therapy with oxygen concentrators. AB - Domiciliary long-term oxygen therapy (LTOT) is usually supplied by means of oxygen concentrators (OCs). Various factors that determine the efficacy of such a treatment were evaluated. Sixty-three patients, arbitrarily selected from lists of health care providers, were visited at home by a biomedical engineer and a pulmonary function technician. The evaluation consisted of: i) responses to a directed questionnaire, ii) assessment of the OC output characteristics, and iii) measurement of the patient's oxygen saturation (SaO2) at rest with and without oxygen supplement. Only 33% of patients received oxygen treatment for the recommended 12-24 hours/day and 5% of patients waited the recommended 10 minutes of OC warm-up before connection. Filters were cleaned weekly by only 30% of patients and the concentrator was serviced 3-4 times a year in 25% of cases. The OC was thought to be unduly noisy by 24% of patients and connecting tubing of less than 6 meters was fitted to 90% of OCs, thereby limiting patient mobility. Most of the OCs did not yield the recommended oxygen concentration and the flow rate meters on them tended to underread. Therefore, only 22% of patients received the prescribed oxygen supplement. Whilst breathing room air, a substantial proportion of patients had an SaO2 >90%. Improvements are clearly required in terms of medical indications for LTOT, patient education and supervision, supply and maintenance of concentrators and related equipment. PMID- 9203515 TI - Lymphocytotoxic antibodies in Israeli patients with rheumatoid arthritis. AB - The presence, antigenic specificity, and clinical role of lymphocytotoxic antibodies (LCAs) were studied in 72 Israeli patients with rheumatoid arthritis (RA). Using the microlymphocytotoxicity assay on a cell panel of 47 donors, LCAs were found in 55% of the 72 RA sera, each displaying a distinct pattern of anti lymphocytic reactivity, mostly against B lymphocytes. Human lymphocytic antigens (HLA) analysis of donors' lymphocytes suggested that activity of LCA-positive RA sera is HLA directed in 60% of cytotoxic sera. The anti-HLA antibodies found were not autoreactive and were not restricted to a particular class I or class II antigen. Relating the presence of LCAs to selective clinical features revealed that LCAs are inversely associated with the presence of an erosive disease (P <0.01) and with the patients' HLA-DQw1 (P <0.01). These findings suggest that LCAs in Israeli patients with RA are very common, multispecific and may have a protective role not mediated through interaction with self-HLA antigens. PMID- 9203516 TI - Proposed therapeutic algorithm for the treatment of anemia of chronic renal failure in pre-dialysis patients with low dose once weekly subcutaneous r-HuEPO. Multicenter Study Group, Israel. AB - Anemia of chronic renal failure (CRF) prior to initiation of dialysis is an important cause of morbidity and requires early therapeutic intervention. The current study was designed to investigate the efficacy and tolerability of a therapeutic algorithm for anemia of CRF in pre-dialysis patients which is based on low dose once-a-week subcutaneous (s.c.) administration of recombinant human erythropoietin (r-HuEPO). Thirty-one patients participated in a prospective open label multicenter study. At baseline, hemoglobin was 8.8+/-0.1 g/dl, transferrin saturation 27+/-2%, ferritin 207+/-28 ng/ml and serum creatinine 4.7+/-0.2 mg/dl. Treatment with r-HuEPO was started at a fixed s.c. dose of 4,000 units once weekly, irrespective of body weight, and titrated upwards or downwards according to a predetermined algorithm. Hemoglobin rose to levels >10 g/dl within 8 weeks and remained stable throughout the remaining period of the study. By week 24, most patients required DAMGO > Dyn-A. Naloxone blocked the inhibitory effects of enkephalins and other opioid agonists on the GABA current. Naltrindole (NTI), a delta-receptor antagonist, prevented the DPDPE-induced depression of the GABA current. beta-Funaltrexamine (beta-FNA), a mu-receptor antagonist, reduced the DAMGO-induced depression of GABA currents. Nor binaltorphimine (nor-BNI), a kappa-receptor antagonist, reduced the effects of Dyn-A in depressing the GABA current. The results suggest that enkephalin down regulates GABA(A) receptor function through mainly delta- and mu-opioid receptors in bullfrog DRG neurons. PMID- 9203546 TI - Effects of age and sociosexual experience on the morphology and metabolic capacity of brain nuclei in the leopard gecko (Eublepharis macularius), a lizard with temperature-dependent sex determination. AB - In vertebrates having sex chromosomes, sexual behavior is influenced by steroid hormones throughout life as well as by the cumulative experiences of the individual. Because males and females differ genetically as well as hormonally, it would be valuable to distinguish the contribution of sex-specific genes from hormones. In addition, since animals age as they gain sociosexual experience, but do not necessarily gain sociosexual experience as they age, it is important to separate the effects of age from those attributable to experience. The leopard gecko is a lizard lacking sex chromosomes, depending instead upon the temperature during incubation to establish gonadal sex. This effectively removes sex-specific genetic influences from any study of sexual differentiation. Eggs were incubated at either 26 degrees C or 32.5 degrees C, temperatures that produce only female hatchlings or a male-biased sex ratio, respectively. By raising geckoes in isolation and then housing some animals together in breeding groups at different ages after they attained sexual maturity, it was possible to assess the relative effects of age and sociosexual experience on the volume and metabolic capacity of limbic and non-limbic brain areas. In general, males showed more changes compared to females. For example, there was a decrease with age in the volume of the preoptic area and the ventromedial hypothalamus in males, but not in females. Both age and sociosexual experience influenced cytochrome oxidase activity in these and other brain areas. Experienced animals had greater metabolic capacity in nuclei functionally associated with sociosexual behavior in lizards and other vertebrates. For example, cytochrome oxidase activity was higher in the anterior hypothalamus of males, in the ventromedial hypothalamus of both males and females from the male-biased incubation temperature, and in the preoptic area of females from both incubation temperatures. These differences were not paralleled by differences in circulating levels of sex hormones; only plasma androgen levels differed as a function of experience in males. These data suggest that the volume and metabolic capacity of specific brain regions change as animals age and gain sociosexual experience, but the nature and degree of change depend upon prenatal events. PMID- 9203547 TI - Neonatal ablations of the amygdala and inferior temporal cortex alter the vocal response to social separation in rhesus macaques. AB - Rhesus macaques that had received bilateral ablations to either the amygdala or area TE in inferior temporal cortex in the 1st week of life were briefly separated from familiar conspecifics at 10-14.5 months of age in order to assess the vocal response to this mild challenge. Sound spectrograms were subjected to quantitative analysis and compared with calls from normal, age-matched controls subjected to the same testing conditions. Animals with TE damage called at a higher rate than animals in the other two groups. TE subjects also produced more coos than controls. Males with TE lesions produced noisy calls at a higher rate than males of the other two groups. Females did not differ between groups in this measure. Analysis of the detailed acoustic structure of the 'coo' indicated significant differences in a measure of slope of the fundamental frequency (rate of frequency change over time) between amygdalectomized animals and those of the other 2 groups. The amygdalectomized monkeys produced calls with lower slope values, giving the calls a less inflected quality both in sonagrams and to the listener. These findings suggest an important role for the amygdala and inferior temporal cortex in regulating the vocal response to social separation during development. PMID- 9203548 TI - Evidence for recovery of spatial learning following entorhinal cortex lesions in mice. AB - The influence of entorhinal cortex lesions on behaviour and concommitant changes in synaptophysin immunoreactivity (IR) in the denervated dentate gyrus was assessed. Male, C57/B6 mice received either bilateral (BI), unilateral (UNI), or no lesion (SHAM) to the entorhinal cortex. At various stages post-lesion the animals were evaluated in tests to examine neurological and cognitive (spatial and cued learning, Morris water maze) function. UNI lesioned animals from 6-36 days post-lesion showed no neurological nor marked cued learning deficit, yet a profound spatial learning deficit. However by 70 days post-lesion, spatial learning ability was clearly evident. In contrast, BI lesioned animals showed severe spatial learning deficits throughout the test period (6-70 days), cued learning was also impaired. In parallel groups of UNI lesioned mice, 6-36 days post-lesion there was a marked reduction (-40%) in synaptophysin IR in the dentate gyrus molecular layer. However by 70 days post-lesion a clear increase in this measure was noted. Changes in the expression of the growth associated protein, GAP43, were also noted over this period. Taken together, the present results suggest some recovery of spatial learning following unilateral entorhinal cortex lesions in mice. This behavioural recovery of a hippocampally dependant task may be associated with a recovery of function related to the synaptic remodelling and elevation of synapse number in the denervated hippocampus, as evidenced by changes in synaptophysin and GAP43 IR. PMID- 9203549 TI - Mapping of carotid baroreceptor subtype projections to the nucleus tractus solitarius using c-fos immunohistochemistry. AB - This study has combined physiological pressure stimulation of carotid baroreceptors via a vascularly isolated carotid sinus and anodal block of baroreceptor afferent fibers in the carotid sinus nerve to examine the medullary projections of type I vs. type II (large A- vs. small A- and C-fiber afferent) baroreceptors. The control distribution of cells in the nucleus tractus solitarius expressing c-fos in response to physiological activation of carotid baroreceptors in the isolated sinus was compared to that during anodal block of large A-fibers in the carotid sinus nerve. Carotid baroreceptor stimulation primarily activated neurons in the ipsilateral commissural and medial subnuclei of the caudal nucleus tractus solitarius and the dorsolateral, dorsomedial and medial subnuclei in the intermediate and rostral levels of the nucleus tractus solitarius. Elimination of large A-fiber carotid baroreceptor afferents, during similar carotid baroreceptor stimulation resulted in a decrease in the number of cells expressing c-fos in the dorsomedial subnucleus of the rostral nucleus tractus solitarius. These data indicate that projections of larger A-fiber (type I) carotid baroreceptors are localized primarily to the rostral dorsomedial subnucleus, while those of smaller A- and C-fiber baroreceptors are more widely distributed to the commissural, medial and dorsal subnuclei of the nucleus tractus solitarius. PMID- 9203551 TI - Medullary post-inspiratory neuronal and diaphragm post-inspiratory activities during spontaneous augmented breaths in anesthetized rats. AB - To test the hypothesis that during augmented breaths the prolonged expiratory time is correlated with the activity of medullary post-inspiratory neurons, changes in diaphragm electromyogram and post-inspiratory neuronal activities were examined in pentobarbital anesthetized rats. During augmented breaths, the mean total expiratory time significantly increased (from 0.57 +/- 0.02 to 1.20 +/- 0.07 s, P < 0.01), but the mean duration of post-inspiratory inspiratory activity (PIIA) was significantly decreased (from 0.08 +/- 0.01 to 0.02 +/- 0.003 s, P < 0.01) compared to control breaths. The mean time which the diaphragm was electrically silent was observed to be significantly prolonged during augmented breaths (from 0.48 +/- 0.02 to 1.19 +/- 0.07 s, P < 0.01). During significant prolongations of expiratory time in augmented breaths, most of the recorded neurons (9/10) in the medulla showed sustained discharges. The mean duration of discharges was longer than that of PIIA (0.02 +/- 0.003 vs. 0.50 +/- 0.10 s, P < 0.01). The mean firing frequency/breath significantly increased compared to control breaths (9.45 +/- 1.69 vs. 37.6 +/- 7.13 imp/breath, P < 0.01). These results suggested that during augmented breaths, the prolonged expiratory period may not be mediated by persisting post-inspiratory neuronal activity in the medulla. PMID- 9203550 TI - Immunohistochemical analysis of presenilin 2 expression in the mouse brain: distribution pattern and co-localization with presenilin 1 protein. AB - Missense mutations of presenilin 1 (PS-1) and presenilin 2 (PS-2) genes cause the majority of early-onset familial forms of Alzheimer's disease (AD). We previously characterized the distribution of the PS-1 protein in the mouse brain by immunohistochemistry using an antibody directed against an epitope located in the large hydrophilic loop [Moussaoui, S., Czech, C., Pradier, L., Blanchard, V., Bonici, B., Gohin, M., Imperato, A. and Revah, F., Immunohistochemical analysis of presenilin 1 expression in the mouse brain, FEBS Lett., 383 (1996) 219-222]. Similarly, we now report the distribution pattern of PS-2 protein in the mouse brain. For these experiments we used a polyclonal antibody raised against a synthetic peptide corresponding to the amino-acid sequence 7-24 of the predicted human PS-2 protein. The specificity of the antibody was evidenced by its ability to recognize PS-2 protein in immunoprecipitation studies and by antigen-peptide competition. In the mouse brain, PS-2 protein was present in numerous cerebral structures, but its distribution in these structures did not correlate with their susceptibility to AD pathology. In all examined structures of the gray matter, PS 2 protein was concentrated in neuronal cell bodies but it was not detected in the glial cells of the white matter. The regional distribution pattern of PS-2 protein was almost identical to that of PS-1 protein. Moreover, PS-2 protein co localized with PS-1 protein in a large number of neuronal cell bodies. In terms of subcellular localization, PS-2 immunostaining was present almost exclusively in neuronal cell bodies while PS-1 immunostaining was also present in dendrites. This could be explained by the different epitopes of the antibodies and the known proteolytic processing of both presenilins in vivo [Tanzi, R.E., Kovacs, D.M., Kim, T.-W., Moir, R.D., Guenette, S.Y. and Wasco, W., The presenilin genes and their role in early-onset familial Alzheimer's disease, Alzheimer's disease Rev., 1 (1996) 91-98]. Within neuronal cell bodies, the immunostaining of PS-2 protein, as well as that of PS-1 protein, had a reticular and granular appearance. This suggests in agreement with previous observations on PS-1 and PS-2 in COS and H4 cells [Kovacs, D.M., Fausett, H.J., Page, K.J., Kim, T.-W., Moir, R.D., Merriam, D.E., Hollister, R.D., Hallmark, O.G., Mancini, R., Felsenstein, K.M., Hyman, B.T., Tanzi, R.E., Wasco, W., Alzheimer-associated presenilins 1 and 2: neuronal expression in brain and localization to intracellular membranes in mammalian cells, Nature Med., 2 (1996) 224-229] that these proteins are situated in intracytoplasmic organelles, possibly the endoplasmic reticulum and the Golgi complex. PMID- 9203553 TI - Novel environment induced inhibition of corticosterone secretion: physiological evidence for a suprachiasmatic nucleus mediated neuronal hypothalamo-adrenal cortex pathway. AB - Basal plasma ACTH and corticosterone levels are controlled by the suprachiasmatic nucleus (SCN), the site of the circadian pacemaker, resulting in a daily peak in plasma corticosterone and ACTH. The present study was carried out to investigate the mechanisms employed by the biological clock to control these hormones. Novel environment induced changes in plasma ACTH and corticosterone in intact and SCN lesioned animals were employed as experimental approach. Placing intact animals in a new environment results in different plasma corticosterone and ACTH responses depending on the clock time of the stimulus. (1) Novel environment (2 h after onset of darkness (ZT14)) results in a fast decrease followed by an increase in corticosterone. This changing pattern in corticosterone secretion was not accompanied by any change in plasma ACTH, suggesting a direct neuronal control of the adrenal cortex. (2) In contrast, novel environment at 2 h after light onset (ZT2) results in a rapid increase in plasma ACTH. Regression analysis of the relation ACTH-corticosterone before and after stress shows a changed pattern at ZT2, although at that time still no significant correlation between ACTH and corticosterone was detected. AT ZT14 this correlation was only present after stress. (3) SCN lesioning results in low basal ACTH at all circadian times combined with elevated corticosterone levels. Here, a new environment results in an immediate increase in corticosterone without inhibition; ACTH also increases rapidly, but attains lower levels than at ZT2 in intact animals. (4) The present results therefore demonstrate SCN modulating corticosterone secretion by affecting ACTH secretion and changing the sensitivity of the adrenal cortex by means of a neuronal input. PMID- 9203552 TI - Brain [3H]cAMP binding sites are unaltered in depressed suicides, but decreased by antidepressants. AB - Saturation binding of [3H]cAMP to the regulatory subunit of cAMP-dependent protein kinase (PKA) was measured in the soluble fraction of brain samples, obtained at post-mortem, from suicides with a firm retrospective diagnosis of depression and individually matched controls. Suicides were subdivided into those who had been free of antidepressant drugs for at least 3 months and those in whom prescription of antidepressants was clearly documented. In antidepressant-free suicides, we found no significant differences in the number or affinity of [3H]cAMP binding sites in the five regions studied. In antidepressant-treated suicides however, Bmax values were lower in all regions, reaching statistical significance in parietal cortex and amygdala. Kd values for antidepressant treated suicides were significantly higher in parietal cortex, temporal cortex and amygdala. These results suggest the regulatory subunit of PKA is unaltered in depression, but is influenced by antidepressant drugs. PMID- 9203554 TI - Selective reduction of monoamine oxidase A and B in the frontal cortex of subordinate rats. AB - We have previously shown that subordination causes a reduction in the levels of 5 hydroxytryptamine and dopamine selectively in the frontal cortex [6]. These monoamines are catabolised mainly by the enzyme monoamine oxidase (MAO) which exists in two isoforms; MAO-A and MAO-B. The present study was carried out to determine whether there is any change in the activity of these two iso-enzymes induced by subordination and if any such alteration is confined to the frontal cortex. The animal model of dominance-subordination used was a worker-parasite paradigm in male Wistar rats. The enzyme activities were measured in five brain regions, the frontal cortex, entorhinal cortex, hippocampus, hypothalamus and striatum, using kynuramine as the substrate. Clorgyline and L-deprenyl were used in vitro to block the activities of MAO-A and MAO-B, respectively. There was a significant (P < 0.001) reduction in the activity of MAO-A as well as MAO-B selectively in the frontal cortex of the subordinate animals. This finding may suggest a reduced neurotransmitter turnover in the serotonergic and dopaminergic neurons terminating in the frontal cortex. PMID- 9203555 TI - 17alpha-Dihydroequilenin increases hippocampal dendritic spine density of ovariectomized rats. AB - The effects of estradiol and 17alpha-dihydroequilenin on the apical dendrite spine density of pyramidal cells of the CA1 region of rat hippocampus were compared. 17alpha-Dihydroequilenin was as effective as estradiol in increasing spine densities relative to controls. 17alpha-Dihydroequilenin is not uterotrophic like estradiol but does have beneficial effects on the cardiovascular system, suggesting that it may be an effective single-agent hormone replacement therapy to treat menopausal symptoms and reduce chronic disease risk in menopausal women. PMID- 9203556 TI - Microinjection of NMDA into the SCN region mimics the phase shifting effect of light in hamsters. AB - Although there is considerable data that glutamate is the primary transducer of photic information to the circadian clock in the suprachiasmatic nucleus (SCN), the ability of glutamate to mimic the phase-shifting effects of light has yet to be demonstrated in vivo. In the present study, microinjections of the glutamate agonist NMDA directly into the SCN of Syrian hamsters induced significant phase delays at circadian time (CT) 13.5 and phase advances at CT 19. These results support the hypothesis that glutamate is the primary neurotransmitter responsible for the transduction of photic information to the SCN. PMID- 9203557 TI - Neuropeptide FF in the VTA blocks the analgesic effects of both intra-VTA morphine and exposure to stress. AB - It has been shown previously that i.c.v. administration of neuropeptide FF (NPFF) attenuates the analgesic effects of exogeneous and endogeneous opioids. Here, we report that intra-ventral tegmental area (VTA) NPFF pre-treatment blocks the analgesia induced by either intra-VTA morphine or exposure to footshock stress in the formalin test for tonic pain. These findings suggest that NPFF in the VTA may play an adaptive role in regulating the response to stress. PMID- 9203558 TI - Increased expression of the peripheral-type benzodiazepine receptor-isoquinoline carboxamide binding protein mRNA in brain following portacaval anastomosis. AB - Using RT-PCR, gene expression of the peripheral-type benzodiazepine receptor isoquinoline carboxamide-binding protein (PTBR-IBP) was studied in the frontal cortex of rats four weeks following end-to-side portacaval anastomosis, an experimental animal model of hepatic encephalopathy, or sham operation. Portacaval anastomosis resulted in increased expression of PTBR-IBP in frontal cortex and in a concomitant increase in densities (Bmax) of binding sites for the PTBR ligand [3H]PK11195. In view of the findings that the PTBR modulates the synthesis of neurosteroids with high affinity for excitatory and inhibitory neurotransmitter systems in brain, increased expression of these receptors could be implicated in the pathogenesis of hepatic encephalopathy. PMID- 9203559 TI - Immunohistochemical localization of macrophage migration inhibitory factor (MIF) in tanycytes, subcommissural organ and choroid plexus in the rat brain. AB - We investigated the immunohistochemical localization of the macrophage migration inhibitory factor (MIF) in the rat brain. In addition to epithelial ependymal cells lining the ventricular wall, tanycytes in the basomedial hypothalamus were heavily immunostained. The immunoreactive processes of tanycytes made contacts to sinusoidal capillaries and reached the pial surface forming an immuno-positive structure at the floor of the hypothalamus. Other immunoreactive cells contained the subcommissural organ in the roof of the third ventricle and the epithelial lamina of the choroid plexus. The localization of MIF in cells which have contact with cerebrospinal fluid and blood vessels suggests that MIF might play a role as a humoral factor in the brain. PMID- 9203560 TI - Infusion of D-cycloserine into temporal-hippocampal areas and restoration of mnemonic function in rats with disrupted glutamatergic temporal systems. AB - Partial transections of the fiber connections between the temporal cortex and the lateral entorhinal cortex at a site of the white matter corresponding to the perirhinal cortex result in impaired visual memory accompanied by reduced concentrations of glutamate in both the temporal cortex and lateral entorhinal cortex. Intraperitoneal administration of the glycinergic receptor agonist D cycloserine produces complete restoration of memory function, as measured by a brightness discrimination task in rats with temporal cortex/lateral entorhinal cortex transections. The purpose of the present study was to identify in which brain structures the compensatory activity might take place. The results show that infusion of cycloserine into either the temporal cortex or lateral entorhinal cortex fully ameliorated the impairment of temporal cortex/lateral entorhinal cortex lesions, whereas infusion into the hippocampal region caused only a mild improvement of the retention performance. Infusion of cycloserine into the frontal cortex or saline into the temporal cortex or lateral entorhinal cortex had no ameliorating effects on the memory dysfunction of rats bearing temporal cortex/lateral entorhinal cortex transections. It is concluded that the temporal cortex, lateral entorhinal cortex and perirhinal cortex are highly critical in forming visual memory. PMID- 9203561 TI - SA4503, a novel cognitive enhancer with sigma1 receptor agonist properties, facilitates NMDA receptor-dependent learning in mice. AB - The selective sigma1 receptor agonist 1-(3,4-dimethoxyphenethyl)-4-(3-phenyl propyl)piperazine dihydrochloride (SA4503) was reported to reverse the amnesia induced by the muscarinic receptor antagonist scopolamine at sub-mg/kg doses. We examined its effect on the learning impairment induced in mice by the non competitive NMDA receptor antagonist dizocilpine. Learning capacities were evaluated using spontaneous alternation in the Y-maze for spatial working memory, and step-down type passive avoidance. SA4503 (0.03-1 mg/kg s.c.) attenuated the dizocilpine (0.15 mg/kg i.p.)-induced memory deficits following a bell-shaped curve in both tests. These effects of SA4503 were blocked by haloperidol (0.05 mg/kg i.p.), implicating sigma1 receptors. SA4503 also reversed the alternation deficit induced by N(omega)-nitro-L-arginine methyl ester (L-NAME, 100 mg/kg i.p.) at the same dosage, indicating that it acted on working memory through the nitric oxide (NO)-mediated signalling pathway. Furthermore, progesterone (2 mg/kg s.c.) blocked the SA4503 effects in the dizocilpine- and L-NAME-amnesia models, in accordance with the purported neurosteroids/sigma1 receptors interaction. These results demonstrate a promising neurobehavioural profile of SA4503, a ligand equally efficient to reverse the deficit in the glutamatergic as well as in the cholinergic amnesia model. Pertinent informations on the potential mechanism of the anti-amnesic effects of sigma1 receptor ligands were also obtained. PMID- 9203562 TI - The adenosine A1 receptor antagonist BIIP 20 counteracts scopolamine-induced behavioral deficits in the passive avoidance task in the rat. AB - The present study investigated the effects of the putative adenosine A1 receptor antagonist BIIP 20 ((S)-(-)-8-(3-oxocyclopentyl)-1,3-dipropyl-7H-purine-2,6 dione) on counteracting scopolamine-induced behavioral deficits in the rat in the passive avoidance paradigm. A single oral application of BIIP 20 (1 and 3 mg/kg) 90 min before the rats received the noxious stimulus significantly attenuated the scopolamine-induced deficits observed during the retention trial of this task. PMID- 9203563 TI - Essential fatty acid preparation improves biochemical and cognitive functions in experimental allergic encephalomyelitis rats. AB - This study examined the possible effects of a novel mixture of fatty acids, SR-3 (a specific ratio of alpha-linolenic acids), on brain biochemistry and on learning deficits induced by injection of an agent that induces experimental allergic encephalomyelitis. Treatment with SR-3 caused a decrease in myelin and changes in the fatty acid profile of brain synaptosomes, and a learning deficit. Eighteen days of treatment with SR-3 reversed the biochemical and learning deficit significantly, but did not restore them to normal levels. We propose that, most probably, the main action of SR-3 is the modulation of the cholesterol level, which in turn causes the modulation of the fatty acid profile and enhances learning by allowing improved neuronal communication. PMID- 9203564 TI - Activation of c-fos mRNA in the brain by the kappa-opioid receptor agonist enadoline and the NMDA receptor antagonist dizocilpine. AB - Using in situ hybridization, we have shown that the kappa-opioid receptor agonist enadoline (CI-977) and the non-competitive NMDA receptor antagonist dizocilpine (MK-901) induced the immediate early gene c-fos in dorsal medial thalamic nuclei. Dizocilpine, and not enadoline, also induced c-fos in the posterior cingulate cortex and retrosplenial cortex. Enadoline's stimulation of c-fos mRNA was dose and time dependent and completely inhibited by pretreatment with naltrexone. Morphine did not stimulate c-fos in the thalamus. It is suggested that the stimulation of c-fos with enadoline represents a specific kappa-opioid receptor effect. PMID- 9203565 TI - Rizatriptan has central antinociceptive effects against durally evoked responses. AB - The 5-HT(1B/1D) receptor agonist rizatriptan constricts intracranial, extracerebral blood vessels, inhibits neurogenic vasodilation and extravasation in the meninges and is effective clinically against migraine. The present study has investigated whether rizatriptan may also have activity at 5-HT(1B/1D) receptors within the central nervous system (CNS) that contributes to its antimigraine effects. Action potentials evoked by electrical stimulation of the dura-mater were recorded extracellularly from single neurones in the trigeminal nucleus caudalis in anaesthetized rats. Rizatriptan dose dependently inhibited these nociceptive dural responses by up to 63 +/- 9% after 3 mg/kg, i.v. Rizatriptan therefore has central activity which may contribute to its efficacy against migraine headache. PMID- 9203566 TI - Nociception from blood vessels is independent of the sympathetic nervous system under physiological conditions in humans. AB - To test the hypothesis that vascular pain depends on sympathetic drive under physiological conditions we studied the effects of both alpha-adrenoceptor stimulation by noradrenaline and alpha-adrenoceptor blockade by phentolamine on the intensity of physicochemically evoked pain from veins in humans. In seven healthy volunteers, a vascularly isolated hand vein segment was perfused continuously with noradrenaline (6 x 10(-9)-6 x 10(-6) M), or phentolamine (1.24 x 10(-4) M). Pain was evoked by intraluminal electrostimulation or by injection of hyperosmolar saline during control perfusion of isoosmolar saline and after each noradrenaline concentration, as well as after perfusion of phentolamine. Subjects rated pain intensity continuously on an electronically controlled visual analogue scale (VAS) between 0% VAS (no pain) and 100% VAS (tolerance maximum). Intravenous electrostimulation as well as hyperosmolar solutions evoked pain in each subject. The intensity of pain was neither influenced by noradrenaline, nor by phentolamine, so that nociception from blood vessels is unlikely to be modulated by the sympathetic nervous system under physiological conditions in humans. PMID- 9203567 TI - N(omega)-nitro-L-arginine methyl ester protects retinal neurons against N-methyl D-aspartate-induced neurotoxicity in vivo. AB - We investigated whether the inhibition of nitric oxide (NO) synthesis with N(omega)-nitro-L-arginine methyl ester (L-NAME), a competitive inhibitor of NO synthase, affects N-methyl-D-aspartate (NMDA)-induced neurotoxicity in the rat retina in vivo. A single intravitreal injection of NMDA damaged the ganglion cell layer and the inner plexiform layer without affecting the other retinal layers 7 days after injection. Intravitreal injection of (5R,10S)-(+)-5-methyl-10,11 dihydro-5H-dibenzo[a,d]cyclo-hepten-5, 10-imine hydrogen maleate (MK-801) with NMDA significantly reduced NMDA-induced degeneration of the retina. NMDA-induced degeneration was also prevented by intravitreal injection of L-NAME but not of D NAME. The protective effect of L-NAME was antagonized by L-arginine. These results suggest that NO plays an important role in NMDA-induced excitotoxic degeneration in the retina. PMID- 9203568 TI - Modification by bradykinin B2 receptor blockade of protection by pacing against ischaemia-induced arrhythmias. AB - In dogs, rapid cardiac pacing, by way of a pacing electrode in the right ventricle, protects against ventricular arrhythmias when a coronary artery is occluded immediately after cessation of the pacing period. This represents a form of ischaemic preconditioning. The role of bradykinin in mediating the protective effects of rapid cardiac pacing in this model was investigated using a selective antagonist of bradykinin at B2 receptors (icatibant; HOE 140). In the presence of icatibant cardiac pacing (220 beats min(-1)) resulted in more severe ischaemia (as assessed by ST-segment elevation from the pacing electrode at the end of the stimulus) and to a higher incidence of ventricular arrhythmias during the pacing protocol. When the coronary artery was occluded under such conditions the antiarrhythmic protection afforded by cardiac pacing was not seen although other indices of reduced ischaemia severity (epicardial ST-segment mapping; changes in the degree of inhomogeneity of electrical activation within the ischaemic area) were not affected by icatibant treatment. These results suggest that bradykinin is an important trigger mediator involved in the protective effects of cardiac pacing. Whether this is due to the generation of endothelium-derived protective substances (such as nitric oxide and prostacyclin) or whether it results from a direct effect on B2 receptors in cardiac myocytes is unclear. PMID- 9203570 TI - Regulation of blood pressure by L-arginine-nitric oxide pathway within the superior colliculus of rats. AB - Injection into the superior colliculus of anaesthetised rats of the nitric oxide (NO) synthase inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME; 1 micromol), but not its inactive enantiomer N(omega)-nitro-D-arginine methyl ester (D-NAME; 1 micromol), significantly (P < 0.01) increased the mean arterial blood pressure. Injection to the superior colliculus of L-arginine (L-Arg; 1 micromol), the substrate for NO synthase, significantly (P < 0.01) lessened the pressor effect of L-NAME, while D-arginine (D-Arg; 1 micromol) did not affect it. L-Arg (7.5 micromol), but not D-Arg (7.5 micromol) administered at the peak of the pressor response to L-NAME (1 micromol) also partially reversed this pressor response (P < 0.05). These data would suggest that endogenously produced NO acts within the superior colliculus to modulate the arterial blood pressure. PMID- 9203569 TI - The novel anti-migraine agent rizatriptan inhibits neurogenic dural vasodilation and extravasation. AB - These studies in anaesthetised rats showed, using intravital microscopy, that the novel anti-migraine agent, rizatriptan, significantly reduced electrically stimulated dural vasodilation but had no effect on increases in dural vessel diameter produced by exogenous substance P or calcitonin gene-related peptide (CGRP). Rizatriptan also significantly inhibited dural plasma protein extravasation produced by high intensity electrical stimulation of the trigeminal ganglion. We suggest that rizatriptan inhibits the release of sensory neuropeptides from perivascular trigeminal nerves to prevent neurogenic vasodilation and extravasation in the dura mater. These prejunctional inhibitory effects may be involved in the anti-migraine action of rizatriptan. PMID- 9203571 TI - Attenuation of endothelium-dependent hyperpolarizing factor by bacterial lipopolysaccharides. AB - Endothelium-dependent hyperpolarizing factor (EDHF) is an important contributor to agonist-induced vascular dilation. Recent studies suggest that bacterial lipopolysaccharides attenuate endothelium-dependent dilation. Whether or not this effect is mediated through inhibition of EDHF is not known. We studied the in vitro influence of Escherichia coli lipopolysaccharides on endothelium-dependent smooth muscle dilation and hyperpolarization in porcine coronary arteries. Endothelium-intact porcine coronary arterial rings were examined after 20 h of incubation with either saline or E. coli lipopolysaccharides (100 microg/ml). Endothelium-dependent dilation elicited by increasing concentrations of bradykinin was significantly attenuated by lipopolysaccharides. Baseline values of smooth muscle membrane potential were not influenced by lipopolysaccharides. However, lipopolysaccharides significantly attenuated bradykinin-induced smooth muscle membrane hyperpolarization. Our results suggest that attenuation of EDHF is an important mechanism through which lipopolysaccharides influence vascular dilation in severe sepsis. PMID- 9203572 TI - Apafant (a PAF receptor antagonist) suppresses the early and late airway responses in guinea pigs: a comparison with antiasthmatic drugs. AB - We studied the effects of apafant (WEB 2086 BS), a specific and potent platelet activating factor (PAF) receptor antagonist, on the early and late airway responses in conscious and actively sensitized guinea pigs. An increase in airway resistance (Rs) was seen 1 min after the inhaled antigen challenge (early airway response), followed by another increase in Rs which peaked between 4 and 8 h after the provocation (late airway response). Oral administration of apafant as well as theophylline inhibited both early and late airway responses. Ozagrel, an inhibitor of thromboxane A2 synthetase, salbutamol, a beta2-adrenoceptor agonist, and dexamethasone significantly inhibited either the early or the late airway response only. Disodium cromoglycate inhibited neither the early nor the late airway response. The results showed that apafant inhibited both the early and late airway responses in sensitized guinea pigs and its effect was comparable or superior to that of anti-asthmatic drugs used clinically. PMID- 9203573 TI - Complex interactions between nitric oxide and adenosine receptors in the rat isolated nodose ganglion. AB - The present study has employed in vitro electrophysiology, utilising the isolated rat nodose ganglion preparation, to determine whether nitric oxide (NO) and adenosine interact with each other in vagal afferent neurons. The nucleophile NO donor, diethylamine-NO, caused reproducible, concentration-related depolarisations of the isolated rat nodose ganglia. Pre-incubation of the isolated rat nodose ganglia with the adenosine A2A receptor agonists CGS 21680 (2 p-(2-carboxyethyl)phenethylamino-5'-N-ethylcarboxamidoadenosine hydrochloride) and DPMA (N6-[2-(3,5-dimethoxyphenyl)-2-(2-methylphenyl)-ethyl]adenosine) (both 1 microM) resulted in a functional antagonism of the ability of diethylamine-NO to depolarise the preparation. A similar effect was observed with adenosine (10 microM) only in the presence of the adenosine A1 receptor antagonist PACPX (1,3 dipropyl-8-(2-amino-4-chlorophenyl)-xanthine, 100 nM). Conversely, the adenosine A1 receptor agonists ENBA (N6-[2-endo-norbomyl]adenosine, 1 microM) and cyclohexyladenosine (100 nM) potentiated the effect of diethylamine-NO on isolated rat nodose ganglia. Inclusion of either adenosine A3 agonists or ATP had no effect on the diethylamine-NO concentration-response curve. These data suggest an ability of NO to interact, in opposing manner, with adenosine A2A and A1 receptors in rat vagal afferent neurons. On the other hand, neither A3 receptors nor ATP appear capable of interacting with NO. PMID- 9203574 TI - Human mast cell tryptase: a stimulus of microvascular leakage and mast cell activation. AB - We have investigated the potential of tryptase to stimulate an increase in microvascular permeability following injection into the skin of guinea pigs. Tryptase was isolated from high salt extracts of human lung tissue by octyl agarose and heparin-agarose chromatography. Injection of purified tryptase (2.5 ng-2.5 microg/site) into the skin of guinea pigs which had been injected intravenously with Evans blue dye provoked a dose-dependent increase in microvascular permeability. The skin reactions elicited by tryptase were apparent up to 80 min following injection, while histamine-induced microvascular leakage resolved completely by 40 min. Heat-inactivation of tryptase, or preincubating the proteinase with certain proteinase inhibitors, significantly reduced the extent of microvascular leakage, suggesting dependency on an intact catalytic site. No evidence was found for a synergistic or antagonistic interaction between tryptase (2.5 ng-2.5 microg/site) and histamine (1-10 microg/site) when these mast cell products were injected together. Addition of heparin to tryptase (10:1; w/w) prior to injection was without effect on tryptase-induced microvascular leakage. Pretreatment of guinea pigs with a combination of the histamine H1 receptor antagonist pyrilamine and the histamine H2 receptor antagonist cimetidine (both 10 mg/kg), partially abolished tryptase-induced microvascular leakage as well as attenuating the reaction to histamine. Reasoning that the microvascular leakage induced by tryptase is likely to involve the release of histamine, we investigated the ability of tryptase to stimulate histamine release from dispersed guinea-pig skin and lung cells in vitro. Tryptase was found to induce concentration-dependent histamine release from both sources of tissue. Mast cell activation stimulated by tryptase in vitro was inhibited by heat treating the enzyme or by addition of proteinase inhibitors, suggesting a requirement for an intact catalytic site. Histamine release was inhibited also by preincubating cells with the metabolic inhibitors antimycin A and 2-deoxy-D glucose indicating that the mechanism was energy-requiring and non-cytotoxic. We conclude that human mast cell tryptase may be a potent stimulus of microvascular leakage. The activation of mast cells by this proteinase may represent an amplification process in allergic inflammation. PMID- 9203575 TI - Cyproterone acetate displaces opiate binding in mouse brain. AB - Drugs acting on androgen receptors modify opioid transmission in the central nervous system. To investigate a direct interaction, we studied whether the binding of [3H]diprenorphine to mouse brain membranes was modified by cyproterone acetate (progesterone derivative with antiandrogen activity), flutamide (non steroidal antiandrogen), 5alpha-dihydrotestosterone and progesterone. Only cyproterone acetate inhibited [3H]diprenorphine binding (IC50 = (1.62 +/- 0.33) x 10(-6) M) without modifying its association rate. These results suggest that cyproterone acetate binds to opiate receptors independently of its classical androgenic intracellular receptor effect. PMID- 9203576 TI - Heterologous desensitization of beta-adrenoceptor signal transduction in vivo. AB - The hemodynamic actions of pituitary adenylate cyclase-activating polypeptide (PACAP-27) rapidly diminish upon repeated i.v. injection in rats treated with the nitric oxide synthase inhibitor, N(G)-nitro-L-arginine methyl ester (L-NAME). We now report that the beta-adrenoceptor agonist isoproterenol (0.5 microg/kg, i.v.) produces pronounced hypotensive and vasodilator effects in anesthetized rats pretreated with L-NAME (100 micromol/kg, i.v.). However, the hypotensive and vasodilator actions of isoproterenol were markedly diminished in L-NAME-treated rats in which tachyphylaxis to PACAP was induced immediately prior to the injection of the beta-adrenoceptor agonist. This suggests that a reduction in tissue concentrations of nitric oxide-containing factors allows tachyphylaxis to PACAP-27-mediated vasodilation to occur in vivo and that this process leads to the heterologous desensitization of beta-adrenoceptor-mediated responses. PMID- 9203577 TI - An activating mutation in a Caenorhabditis elegans Gs protein induces neural degeneration. AB - Heterotrimeric guanine nucleotide-binding proteins (G proteins) act as signal transducing molecules that connect serpentine-transmembrane receptors to a variety of intracellular effectors. We characterized a Caenorhabditis elegans G(s) gene, gsa-1, which encodes a G(s) alpha-subunit (G alpha(s)) that is expressed throughout the nervous system and in muscle cells. gsa-1 is an essential gene; a loss-of-function mutation in gsa-1 results in lethality at the first stage of larval development. Partial (mosaic) loss of G alpha(s) expression or overexpression of the protein results in reciprocal defects in movement and egg-laying, suggesting a role for G alpha(s) in the regulation of these behaviors. Expression of a constitutively active form of G alpha(s) from an inducible promotor results in hypercontraction of body-wall muscle cells and vacuolization and degeneration of neurons within hours of induction. Neurons that are susceptible to the degeneration induced by activated G alpha(s) are predominantly motoneurons located within the ventral nerve cord. Phenotypic analysis shows that the induced neural degeneration is not the result of programmed cell death but is probably caused by the activation of ion channels. A genetic suppressor of activated G alpha(s) was isolated that identifies a putative downstream target of G(s) signaling. PMID- 9203578 TI - ORC-dependent and origin-specific initiation of DNA replication at defined foci in isolated yeast nuclei. AB - We describe an in vitro replication assay from yeast in which the addition of intact nuclei to an S-phase nuclear extract results in the incorporation of deoxynucleotides into genomic DNA at spatially discrete foci. When BrdUTP is substituted for dTTP, part of the newly synthesized DNA shifts to a density on CsCl gradients, indicative of semiconservative replication. Initiation occurs in an origin-specific manner and can be detected in G1- or S-phase nuclei, but not in G2-phase or mitotic nuclei. The S-phase extract contains a heat- and 6-DMAP sensitive component necessary to promote replication in G1-phase nuclei. Replication of nuclear DNA is blocked at the restrictive temperature in an orc2-1 mutant, and the inactive Orc2p cannot be complemented in trans by an extract containing wild-type ORC. The initiation of DNA replication in cln-deficient nuclei blocked in G1 indicates that the ORC-dependent prereplication complex is formed before Start. This represents the first nonviral and nonembryonic replication system in which DNA replication initiates in an ORC-dependent and origin-specific manner in vitro. PMID- 9203579 TI - The Spg1p GTPase is an essential, dosage-dependent inducer of septum formation in Schizosaccharomyces pombe. AB - The spg1 gene (septum-promoting GTPase) was cloned as a multicopy suppressor of a dominant-negative mutant of the Cdc7p kinase. It encodes a small GTPase of the Ras superfamily. spg1 is an essential gene. Null or heat-sensitive alleles do not make a division septum, but growth, S-phase, and mitosis continue in the absence of cell division, producing elongated, multinucleate cells. Increased expression of Spg1p induces septum formation in G2, S-phase, and pre-Start G1-arrested cells. This requires the activity of Cdc7p kinase, but not p34(cdc2). Increased expression of Cdc7p bypasses the requirement for Spg1p. Spg1p and Cdc7p can be coimmunoprecipitated from cell extracts, and interact in the two-hybrid system. These data indicate that Spg1p is a key element in controlling the onset of septum formation in Schizosaccharomyces pombe, and that it acts through the Cdc7p kinase. PMID- 9203580 TI - Nedd5, a mammalian septin, is a novel cytoskeletal component interacting with actin-based structures. AB - The mouse Nedd5 gene encodes a 41.5-kD GTPase similar to the Saccharomyces and Drosophila septins essential for cytokinesis. Nedd5 accumulates near the contractile ring from anaphase through telophase, and finally condenses into the midbody. Microinjection of anti-Nedd5 antibody interferes with cytokinesis, giving rise to binucleated cells. In interphase and postmitotic cells, Nedd5 localizes to fibrous or granular structures depending on the growth state of the cell. The Nedd5-containing fibers are disrupted by microinjection of GTPgammaS and by Nedd5 mutants lacking GTP-binding activity, implying that GTP hydrolysis is required for its assembly. The Nedd5-containing fibers also appear to physically contact actin bundles and focal adhesion complexes and are disrupted by cytochalasin D, C3 exoenzyme, and serum starvation, suggesting a functional interaction with the actin-based cytoskeletal systems in interphase cells. PMID- 9203581 TI - Fission yeast WD-repeat protein pop1 regulates genome ploidy through ubiquitin proteasome-mediated degradation of the CDK inhibitor Rum1 and the S-phase initiator Cdc18. AB - In fission yeast, maintenance of genome ploidy is controlled by at least two mechanisms. One operates through the Cdc2/Cdc13 kinase, which also involves the CDK inhibitor Rum1, and the other through the S-phase regulator Cdc18. By screening for sterile mutants that show increased ploidy, we have identified a new gene, pop1+, in mutants that become polyploid. The pop1 mutation shows a synthetic lethal interaction with the temperature-sensitive cdc2 or cdc13 mutation. In a pop1 mutant Rum1 and Cdc18 proteins become accumulated to high levels. The high ploidy phenotype in the pop1 mutant is dependent on the presence of the rum1+ gene, whereas the accumulation of Cdc18 is independent of Rum1. The predicted sequence of the Pop1 protein indicates that it belongs to a WD-repeat family with highest homology to budding yeast Cdc4, which participates in the ubiquitin-dependent pathway. Consistent with this notion, in a mutant of the 26S proteasome, higher molecular weight forms of Rum1 and Cdc18 are accumulated corresponding to polyubiquitination of these proteins. In the pop1 mutant, however, no ubiquitinated forms of these proteins are detected. Finally we show that Pop1 binds Cdc18 in vivo. We propose that Pop1 functions as a recognition factor for Rum1 and Cdc18, which are subsequently ubiquitinated and targeted to the 26S proteasome for degradation. PMID- 9203582 TI - ClpX and MuB interact with overlapping regions of Mu transposase: implications for control of the transposition pathway. AB - Transposition of phage Mu is catalyzed by an extremely stable transposase-DNA complex. Once recombination is complete, the Escherichia coli ClpX protein, a member of the Clp/Hsp100 chaperone family, initiates disassembly of the complex for phage DNA replication to commence. To understand how the transition between recombination and replication is controlled, we investigated how transposase-DNA complexes are recognized by ClpX. We find that a 10-amino-acid peptide from the carboxy-terminal domain of transposase is required for its recognition by ClpX. This short, positively charged peptide is also sufficient to convert a heterologous protein into a ClpX substrate. The region of transposase that interacts with the transposition activator, MuB protein, is also defined further and found to overlap with that recognized by ClpX. As a consequence, MuB inhibits disassembly of several transposase-DNA complexes that are intermediates in recombination. This ability of MuB to block access to transposase suggests a mechanism for restricting ClpX-mediated remodeling to the proper stage during replicative transposition. We propose that overlap of sequences involved in subunit interactions and those that target a protein for remodeling or destruction may be a useful design for proteins that function in pathways where remodeling or degradation must be regulated. PMID- 9203583 TI - Mlh1 is unique among mismatch repair proteins in its ability to promote crossing over during meiosis. AB - In eukaryotes, homologs of the bacterial MutS and MutL proteins function in DNA mismatch repair and recombination pathways. The mutL homolog MLH1 is required for nuclear mismatch repair. Previously, cytological analysis of MLH1-deficient mice has implied a role for Mlh1 in crossing-over during meiosis. Here we demonstrate that Saccharomyces cerevisiae diploids containing a deletion of MLH1 have reduced crossing-over in addition to a deficiency in the repair of mismatched DNA during meiosis. Absence of either of the meiosis-specific mutS homologs Msh4 or Msh5 results in a similar reduction in crossing-over. Analysis of an mlh1 msh4 double mutant suggests that both genes act in the same pathway to promote crossing-over. All genetic markers analyzed in mlh1 mutants display elevated frequencies of non Mendelian segregation. Most of these events are postmeiotic segregations that represent unrepaired heteroduplex. These data suggest that either restorational repair is frequent or heteroduplex tracts are shorter in wild-type cells. Comparison of mlh1 segregation data with that of pms1, msh2, msh3, and msh6 mutants show that the ability to promote crossing-over is unique to MLH1. Taken together these observations indicate that both crossing-over and gene conversion require MutS and MutL functions and that Mlh1 represents an overlap between these two pathways. Models of Mlh1 function are discussed. PMID- 9203584 TI - The mesenchymal winged helix transcription factor Fkh6 is required for the control of gastrointestinal proliferation and differentiation. AB - The winged helix transcription factor Fkh6 is expressed in the mesoderm of the gastrointestinal tract directly adjacent to the endoderm-derived epithelium. Homozygous null mice for Fkh6 showed postnatal growth retardation secondary to severe structural abnormalities of the stomach, duodenum, and jejunum. Dysregulation of epithelial cell proliferation in these organs resulted in an approximately fourfold increase in the number of dividing intestinal epithelial cells and marked expansion of the proliferative zone. As a consequence, the tissue architecture of the stomach and small intestine was distorted, with abnormal crypt structure, formation of mucin filled cysts, and lengthening of villi. Changes in the cellular phenotype and composition of the gastric and intestinal epithelia also suggests that epithelial cell-lineage allocation or differentiation may be affected by loss of Fkh6. From the analysis of a number of potential signaling molecules, we found Bmp2 and Bmp4 expression reduced in the gastrointestinal tract of Fkh6 mutant mice, suggesting that Fkh6 directs a signaling cascade that mediates communication between the mesenchyme and endoderm of the gut to regulate cell proliferation. PMID- 9203585 TI - Deletion of the H19 transcription unit reveals the existence of a putative imprinting control element. AB - The distal region of mouse chromosome 7 contains a cluster of imprinted genes that includes H19 and Igf2 (insulin-like growth factor 2). H19 is expressed as an untranslated RNA found at high levels in endodermal and mesodermal embryonic tissues. This gene is imprinted and exclusively expressed from the allele of maternal origin. The Igf2 gene shows a similar pattern of expression but is expressed from the paternal allele. We have generated a targeted deletion of the H19 transcription unit by insertion of a neo replacement cassette. The homozygous mutant animals are viable and fertile and display an overgrowth phenotype of 8% compared with wild-type littermates. This is associated with the disruption of Igf2 imprinting and the consequent biallelic expression of this gene. A striking feature of the recombinant H19 allele is the occurrence of a parental imprint set on the neo replacement cassette. Therefore imprinting of the H19 locus is independent of the H19 gene itself. Taken together with the results of a larger H19 mutation described previously, this indicates that an imprinting control element is located within the region 10 kb upstream of H19. PMID- 9203586 TI - SV40 large T antigen binds to the TBP-TAF(I) complex SL1 and coactivates ribosomal RNA transcription. AB - SV40 large T antigen is a multifunctional regulatory protein that plays a key role in the viral life cycle and can stimulate cell proliferation. To accomplish this, large T antigen has to control the expression of cellular genes involved in cell cycle progression and cell growth. rRNA synthesis by RNA polymerase I (Pol I) is tightly associated with cell growth and proliferation, and previous studies indicated that large T antigen up-regulates RNA Pol I transcription in SV40 infected cells. How this process occurs is currently unclear. To investigate the mechanisms of large T antigen stimulation of RNA Pol I transcription, we have established an in vitro transcription system that is responsive to large T antigen. Here, we show that recombinant large T antigen stimulates Pol I transcription reconstituted with purified RNA Pol I, UBF, and the TBP/TAF complex SL1. Immunoprecipitation experiments revealed that large T antigen directly binds to SL1, in vitro, as well as in SV40-infected cells. In addition, our data indicate that this interaction occurs by direct association with three SL1 subunits, namely TBP, TAF(I)48, and TAF(I)110. Transcription studies with large T antigen deletion mutants show that the 538-amino-acid amino-terminal domain is necessary for full stimulation of Pol I transcription. Importantly, mutants that no longer bind to SL1 are also unable to stimulate Pol I transcription. This indicates that recruitment of large T antigen to the rRNA promoter by SL1 constitutes a crucial step in the activation process. Taken together with recent studies on large T antigen activation of RNA Pol II transcription, these results suggest that viral modulation of genes involved in cell proliferation involves direct targeting of promoter-specific TBP/TAF complexes (i.e., SL1 or TFIID) by large T antigen. PMID- 9203587 TI - Minimally invasive coronary artery bypass grafting: on the beating heart and via limited access. AB - Minimally invasive coronary artery bypass grafting (MICABG) may be achieved by arterial grafting on the beating heart, without cardiopulmonary bypass, and by operations via limited access. The Second Utrecht MICABG Workshop held October 4 5, 1996, focused on beating-heart coronary immobilization, limited-access thoracoscopic and direct-vision mobilization of the internal mammary artery, limited-access left anterior descending coronary artery grafting, and, finally, facilitated distal anastomosis techniques. It has yielded 33 reports in this supplement. The combined, cumulative experience of a number of participants exceeded 3,000 beating-heart cases, including more than 1,000 with arterial grafting through limited access. The average number of anastomoses per patient ranged from 1.0 to 2.0. Therapeutic strategies are evolving, and dedicated instrumentation is being developed. Randomized clinical trials with angiographic follow-up are required to establish that the reduction in invasiveness of coronary bypass grafting is not achieved at the expense of suboptimal quality of the arterial graft and the distal anastomosis. PMID- 9203588 TI - Clinical assessment of functional stenosis severity: use of coronary pressure measurements for the decision to bypass a lesion. AB - BACKGROUND: In the selection of patients eligible for minimally invasive coronary artery bypass grafting (MICABG), knowledge about the pathophysiologic significance of individual coronary stenoses is important. Only if the lesion amenable to MICABG can be identified as the culprit lesion, and other lesions can be demonstrated not to be responsible for reversible ischemia, will MICABG be an appropriate procedure. METHODS: By simultaneous measurement of mean aortic pressure and transstenotic coronary pressure, a pathophysiologic index can be obtained that specifically indicates the influence of an epicardial coronary stenosis on maximum achievable blood flow of the supplied myocardial territory. This index is called myocardial fractional flow reserve (FFR(myo)). RESULTS: Myocardial fractional flow reserve is a reliable, lesion-specific index for determining whether a particular stenosis is responsible for reversible myocardial ischemia. If FFR(myo) is less than 0.75, revascularization is indicated, whereas if FFR(myo) is greater than 0.75, revascularization usually is not warranted. Moreover, in contrast to classic coronary flow or flow velocity reserve, FFR(myo) is independent of changes in heart rate, blood pressure, and contractility, and also accounts for the contribution of collaterals. CONCLUSIONS: Pressure-derived FFR(myo) is an accurate pathophysiologic index for reliable identification of functionally significant epicardial lesions and can be obtained easily and quickly during routine cardiac catheterization. Therefore, FFR(myo) facilitates clinical decision-making with respect to the appropriateness of MICABG. PMID- 9203589 TI - Intraoperative monitoring of IMA flow: what does it mean? AB - BACKGROUND: This study examines whether the measurement of internal thoracic artery (ITA) graft flow can determine the adequacy of the ITA-left anterior descending coronary artery (LAD) anastomosis. METHODS: To study a wide range of clinical problems, we used a computer simulation of the cardiovascular system. The model included a time-varying elastance model of the heart, a systemic circulation represented by a multielement nonlinear model of the aorta and its major branches, a nonlinear model of the LAD circulation, and a model of the ITA bypass graft. RESULTS: With a mild LAD stenosis, ITA graft flow was low and flow reversal occurred. As the percent stenosis increased, ITA flow and the percentage of ITA-to-total LAD flow increased. The ITA graft helped to maintain resting LAD blood flow. A partial obstruction (40%) at the ITA-LAD anastomosis reduced ITA graft flow at similar levels of LAD stenosis. However, overlap in flow values comparing a normal with a partially obstructed anastomosis occurred. CONCLUSIONS: Flow patterns in the ITA are highly dependent on the degree of stenosis of the LAD as well as the integrity of the anastomosis. The predictive power of ITA flow measurement increases with severe stenosis or total occlusion of the proximal LAD and with high coronary blood flow demands. PMID- 9203590 TI - Role of myocardial protection for coronary artery bypass grafting on the beating heart. AB - BACKGROUND: Minimal invasive coronary artery bypass grafting is often performed on the warm, beating heart. This setting requires special measures for protection of the myocardium during acute regional ischemia under normothermic conditions. METHODS: A first possibility to enhance the tolerance to ischemia during short episodes of coronary artery occlusion is based on a pharmacologic approach. A second possibility is mechanical unloading of the ischemic heart during and after regional ischemia. RESULTS: To interfere with the ionic imbalances leading to necrosis, blockade of the Na(+)-H+ exchanges can be induced. It is shown that the selective Na(+)-H+ exchange inhibitor HOE 694 is able to prevent ischemic contracture in the experimental setting. Another pharmacologic approach is the use of endogenous adenosine accumulation in the ischemic myocardium. This can be achieved by the use of nucleoside transport inhibitors (lidoflazine or nitrobenzylthioinosine) having the ability to accumulate adenosine at the site of its production. Adenosine has a strong cardioprotective effect via adenosine A1 receptor stimulation and induces bradycardia by opening K+ channels and increasing the potassium current. It is also shown that left ventricular unloading by an axial flow pump (Hemopump) improves postischemic myocardial dysfunction (stunning) in an experimental model of short regional ischemia and reperfusion. CONCLUSIONS: Further clinical investigation of both pharmacologic and mechanical techniques for cardioprotection during minimally invasive coronary artery bypass grafting is required. PMID- 9203591 TI - Revascularization of patients with coronary artery disease: the interventional cardiologist's perspective. AB - In the majority of patients with chronic coronary artery disease, treatment is aimed at palliation or prolongation of disease-free intervals and consists of either pharmacologic therapy or coronary revascularization. As a result of continuous refinements and improvements in both surgical and catheter-based revascularization techniques, modalities, and adjunctive pharmacologic therapy, an increasing number of patients may benefit from coronary revascularization. This also engenders difficult choices for the physicians responsible for selecting the most appropriate treatment. To achieve and provide optimal patient care an open and principled discussion with all parties involved is mandatory and must be based on the integration of clinical experience and data from both basic and clinical research. The purpose of this article is to provide the interventional cardiologist's view on the treatment of patients with atherosclerotic coronary artery disease. PMID- 9203592 TI - Vagus nerve stimulation as a method to temporarily slow or arrest the heart. AB - BACKGROUND: Electrical stimulation of nerves is used to study nervous system and body function relationships. Electrical stimulation of the vagus nerve was used to slow the heart during coronary artery bypass grafting. METHODS: A 48-year-old man with multivessel coronary artery disease, scheduled for revascularization, gave informed consent for the surgeon to stimulate his vagus nerve. As part of the operation the left internal mammary artery was harvested as a pedicle and the patient was placed on cardiopulmonary bypass. The vagus nerve was isolated as it crossed the aorta just lateral to the phrenic nerve. Pacing wires were placed (1 cm apart) allowing prodromic conduction. With the patient fully supported by cardiopulmonary bypass and after administration of neostigmine (2.5 mg intravenously) eight separate continuous 5-second electrical pulse trains (25 Hz, 20 V, pulse width of 0.1 ms) were delivered to the nerve with 30-second rest periods between each stimulation. During the periods of stimulation the mammary artery to left anterior descending artery anastomosis was completed. RESULTS: Electrical stimulation caused cessation of the heartbeat, termination of the same resulted in normal sinus rhythm, although it was slowed by the neostigmine. Suturing of the anastomosis was done during periods of stimulation. Additional anastomoses were completed using cardiopulmonary bypass-delivered cardioplegia and aortic cross-clamping. CONCLUSIONS: Electrical stimulation of the vagus nerve slowed and temporarily arrested the heart for brief periods to allow critical placement of anastomotic sutures. PMID- 9203593 TI - Transient ventricular asystole using adenosine during minimally invasive and open sternotomy coronary artery bypass grafting. AB - BACKGROUND: The emergence of minimally invasive coronary artery bypass grafting and recent off-pump open sternotomy clinical reports have refocused attention on the technical aspects and outcome of grafting on the beating heart. METHODS: To optimize the surgical field we report a method using adenosine for induction of controlled intervals of ventricular asystole to produce a transiently still cardiac field that facilitates anastomotic accuracy. RESULTS: Adenosine was used in 57 patients, 31 included off-pump coronary artery bypass grafting (27 by minimally invasive technique, 4 by open sternotomy). In a further 26 patients adenosine pauses were used for suture placement to control anastomotic bleeding after cardiopulmonary bypass. Average adenosine boluses per anastomosis were 9 (6 14), mean dose of adenosine per bolus (mg/kg) was 0.24 (0.15-0.35), mean duration of pause (seconds) was 6 (3-19), and mean time for arterial blood pressure (mean) to return to baseline (seconds) was 35 (13-48). Presence of repolarization arrhythmias was noted in 1 patient. There were no deaths. Two patients had recurrent myocardial ischemia shown on angiography to be the result of technical problems. CONCLUSIONS: This report describes our experience with the emerging procedure of minimally invasive coronary operations and off-pump grafting with the adenosine technique. The method also includes mechanical devices and other pharmacological therapy to optimize the surgical field, and the technique has now become a standard component of our off-pump revascularization methods. PMID- 9203594 TI - Port-access cardiac operations with cardioplegic arrest. AB - BACKGROUND: A less invasive approach to cardiac surgery has been propelled by recent advances in video-assisted surgery. Previous obstacles to minimally invasive cardiac operations with cardioplegic arrest included limitations in operative exposure, inadequate perfusion technology, and inability to provide myocardial protection. METHODS: Port-access technology allows endovascular aortic occlusion, cardioplegia delivery, and left ventricular decompression. The endoaortic clamp is a triple-lumen catheter with an inflatable balloon at its distal end. Antegrade cardioplegia is delivered through a central lumen, which also acts as an aortic root vent, a second lumen is used as an aortic root pressure monitor, and a third lumen is used for balloon inflation to provide aortic occlusion. RESULTS: Experimental and clinical studies have demonstrated the feasibility of port-access coronary artery bypass grafting and port-access mitral valve procedures. Endovascular cardiopulmonary bypass using the endoaortic clamp was effective in achieving cardiac arrest and myocardial protection to allow internal mammary artery to coronary artery anastomosis in a still and bloodless field. Intracardiac procedures, such as mitral valve replacement or repair, have been successfully performed clinically. CONCLUSION: The port-access system effectively achieves cardiopulmonary bypass and cardioplegic arrest, thereby enabling the surgeon to perform cardiac procedures in a minimally invasive fashion. This system provides for endovascular aortic occlusion, cardioplegia delivery, and left ventricular decompression. PMID- 9203595 TI - Reoperative coronary artery bypass without cardiopulmonary bypass. AB - BACKGROUND: Conventional reoperative coronary artery bypass grafting using cardiopulmonary bypass carries relatively high mortality and morbidity. METHODS: Seventy-seven patients underwent coronary artery bypass grafting without cardiopulmonary bypass in two centers between 1988 and 1994. Mean age was 65 +/- 8 years (mean +/- SD). Twenty-three (30%) were operated on urgently and 7 (9%) emergently. Nine (12%) were referred for operation up to 2 weeks after acute myocardial infarction. Fifteen patients (19%) had an ejection fraction less than or equal to 0.35. The mean number of grafts per patient was 1.7 (range, 1 to 3), and the internal mammary artery was used in 66 patients (86%). Only 18 patients (23%) received at least one graft to the circumflex artery. Hospital stay was 7.4 +/- 6.5 days. RESULTS: Early events included operative death in 4 patients (5.2%), nonfatal myocardial infarction in 3 (3.9%), sternal infection in 2 (2.6%), and stroke in 0 (0%). Follow-up (30 +/- 15 months) showed 11 deaths (5 cardiac, 6 noncardiac), 2 (2.8%) nonfatal myocardial infarctions, and return of angina in 9 patients (12.8%). One- and 4-year actuarial survival rates were 90% and 69%, respectively. CONCLUSIONS: Reoperative coronary artery bypass grafting without cardiopulmonary bypass has acceptable early and midterm outcome, and should be considered a viable alternative for properly selected patients. PMID- 9203596 TI - Primary coronary artery bypass grafting without cardiopulmonary bypass in impaired left ventricular function. AB - BACKGROUND: Conventional coronary artery bypass grafting using cardiopulmonary bypass carries relatively high mortality and morbidity for patients with left ventricular dysfunction. METHODS: Seventy-five patients with ejection fraction less than or equal to 0.35 underwent primary coronary artery bypass grafting without cardiopulmonary bypass between December 1991 and December 1994. Thirty two patients (43%) had congestive heart failure, 11 (15%) were referred for operation within the first 24 hours of evolving myocardial infarction, and 21 (28%) up to 1 week after acute myocardial infarction. Eighteen patients (24%), 6 of whom were in cardiogenic shock, underwent emergency operations. RESULTS: Mean number of grafts/patient was 1.9 (range, 1 to 4), and internal mammary artery was used in 66 patients (85%). Only 17 patients (23%) received a graft to a circumflex marginal artery. Two patients (2.7%) died perioperatively, and 1 (1.3%) sustained stroke. At mean follow-up of 28 months, 13 patients had died, and angina had returned in 7 (10.5%). One- and four-year actuarial survival was 96% and 73%, respectively. CONCLUSIONS: Coronary artery bypass grafting without cardiopulmonary bypass is a viable alternative to conventional coronary artery bypass grafting particularly for patients with extreme left ventricular dysfunction or those with coexisting risk factors, such as acute myocardial infarction and cardiogenic shock. PMID- 9203597 TI - Minimally invasive direct coronary artery bypass grafting: experimental and clinical experiences. AB - BACKGROUND: This communication briefly details the goals, indications, surgical approaches, and limitations of minimally invasive direct coronary artery bypass grafting (MIDCABG). The experimental experiences from various institutions are summarized. METHODS: The clinical experiences of 72 consecutive MIDCABG procedures performed at our institutions between June 5, 1995, and August 13, 1996, were analyzed. We have divided patients into two groups. Group A consists of healthy low-risk patients with single lesions of the left anterior descending coronary artery or the right coronary artery, or with both lesions of both arteries. Group B consists of high-risk patients who had major contraindications to conventional cardiopulmonary bypass procedures. There were 55 patients in group A and 17 patients in group B. Using The Society of Thoracic Surgeons preoperative predicted risk module, group A had a 1% predicted mortality versus 4% in group B. RESULTS: The 30-day mortality was 2% in group A and 6% in group B. The mean postoperative length of stay was 4 days for group A and 5 1/2 days for group B. Short-term follow-up of the survivors appears promising, and 81% of patients were angina free at the time of last follow-up. CONCLUSIONS: The MIDCABG techniques are still developing. The short-term results during the learning period appear to be quite good, but long-term results remain yet to be seen. The addition of new equipment to facilitate construction of the anastomosis will enhance application and results. The lessons learned from these approaches are already being applied to other areas of cardiac surgery including valve replacement and the repair of congenital heart defects. PMID- 9203598 TI - Minimally invasive coronary revascularization through parasternal incisions without cardiopulmonary bypass. AB - BACKGROUND: We report the results of minimally invasive coronary revascularization without cardiopulmonary bypass through miniparasternal incisions. METHODS: This procedure was performed in 40 patients with disease in the left anterior descending, first diagonal, and right coronary arteries. After a 5- to 7-cm left vertical parasternal incision and removal of two costal cartilages, the left internal mammary artery was harvested up to the 2nd rib. The left anterior descending artery was occluded by means of two polydioxanone monofilament sutures. The anastomosis was performed with one 7-0 Prolene suture while the heart was beating. In 4 cases the left internal mammary artery was used as a sequential graft to the left anterior descending artery and the first diagonal artery. In 14 cases the right coronary artery was grafted with the right internal mammary artery through a right parasternal incision. Postoperatively, 95% of the patients underwent angiographic assessment of the anastomoses. RESULTS: We performed 52 anastomoses (34 to the left anterior descending artery, 4 to the first diagonal artery, and 14 to the right coronary artery). The mortality was 0% and the morbidity included postoperative bleeding (5%), acute renal failure (2.5%), atrial fibrillation (2.5%), and wound infection (5%). No patient had ventricular arrhythmias or circulatory problems during or after the operation. Two patients (5%) with right internal mammary artery-to-right coronary artery grafting had graft failure that required a redo operation. CONCLUSIONS: Small vertical parasternal incisions may be an alternative approach for single and multiple coronary revascularization, with a low incidence of intraoperative cardiac complications. The application of this approach to the right coronary artery, however, carries additional technical difficulties, and careful patient selection may be required to achieve optimal results. PMID- 9203599 TI - Coronary bypass grafting via minithoracotomy on the beating heart. AB - BACKGROUND: Recently the availability and the superiority of less invasive coronary artery bypass grafting on some selected groups of patients in the meaning of patient comfort and short hospital stay has been shown by some authors. We present here the clinical results of 40 patients operated on by minithoracotomy incision on the beating heart without using cardiopulmonary bypass mostly harvesting the left internal thoracic artery by videothoracoscopic assistance. METHODS: Between March 1996 and September 1996, 40 patients were operated on by harvesting the left internal thoracic artery mostly by video assisted thoracoscopy and performing bypass through a minitoracotomy incision. Two patients in whom the procedure was switched to conventional technique were not included in this series. Nine of the patients were female and the rest were male. The mean age was 43.2 +/- 7. RESULTS: Left internal thoracic arteries were harvested by video-assisted thoracoscopy completely in 11 patients, incompletely in 24 patients (the harvesting was completed by direct vision afterwards), and under direct vision in 5 through a mini-anterior thoracotomy incision. Thirty-six patients received a bypass graft to left anterior descending coronary artery only, whereas 4 received a diagonal branch graft also. Left internal thoracic arteries were used to bypass the left anterior descending coronary artery directly in 38 patients. The left internal thoracic artery was injured in the middle portion during harvesting in 1 of the remaining 2 patients. The length was not enough in the other. A short saphenous vein graft was interposed between the left internal thoracic artery and the left anterior descending coronary artery in these 2 patients. There was no mortality. One patient had perioperative myocardial infarction. We did not see serious morbidity except one lung injury due to the trochar. CONCLUSIONS: The results obtained from our experience suggest that coronary artery bypass grafting by minithoracotomy could be applied effectively and safely without overwhelming additional risk to the patient. Furthermore, it has some advantages in reducing operative trauma and cost and also improving patient comfort. PMID- 9203600 TI - Minimally invasive coronary artery bypass techniques as adjunct to extracardiac procedures. AB - BACKGROUND: Surgical management of coronary artery disease has improved dramatically over the last decades in terms of short- and long-term results. Nevertheless, elderly patients (more than 75 years); patients with reduced ejection fraction (less than 0.25), heavily calcified aorta, or coexisting noncardiac diseases; and patients requiring cardiac reoperation have an increased perioperative risk when operated on with cardiopulmonary bypass. Successful minimally invasive coronary artery bypass grafting without cardiopulmonary bypass has been reported in selected cases. METHODS: In 8 of 40 high-risk patients undergoing operation on a beating heart, minimally invasive coronary bypass grafting was combined with vascular (carotid endarterectomy, n = 3; aortic replacement, n = 2) and abdominal procedures (a second look after combined pancreas and kidney transplantation) or defibrillator implantations (n = 2). RESULTS: Postoperatively, there was no mortality, no morbidity, and no blood transfusion. Patients are free of symptoms at an average follow-up time of 5.5 +/ 5 months. CONCLUSIONS: Our results indicate that minimally invasive coronary artery bypass grafting technique can be particularly useful if noncardiac procedures have to be performed in high-risk patients with significant coronary artery disease. PMID- 9203601 TI - Anastomotic complications in minimally invasive coronary bypass grafting. AB - BACKGROUND: Anterior wall myocardial revascularization through a left anterior minithoracotomy is an increasingly accepted procedure. Technical failure at the anastomotic site, promoting persistent or recurrent angina, is known to occur and may be underrecognized. This report summarizes the incidence of technical failure in an initial clinical experience and describes potential causes of early postoperative complications. METHODS: Between December 1995 and May 1996, 15 patients underwent left internal mammary artery-to-left anterior descending artery revascularization without extracorporeal circulation. The surgical indication was single-vessel coronary disease in all patients. We exposed the left anterior descending artery target site through a 10-cm left anterior fourth space thoracotomy. The fourth costal cartilage was resected and the left internal mammary artery was harvested under direct visualization. Two 4-0 polypropylene sutures snared in tourniquets proximal and distal to the anastomotic site were used to obtain a bloodless field and stabilization of the left anterior descending artery. RESULTS: All patients had procedures initially deemed successful based on disappearance of angina or postoperative transthoracic Doppler examination of the internal mammary artery 3 to 5 days postoperatively. However, 3 patients presented with recurrent angina at 2, 6, and 8 weeks. Angiography or direct visualization at operation demonstrated the technical complication (stenosis at the anastomotic site in 2 and snare injury in the native vessel in 1). Two patients required reoperation. CONCLUSIONS: Initial results with minimally invasive coronary bypass grafting have generated great enthusiasm worldwide, but there is no consensus on how the procedure should be performed. These results suggest that a nonstabilized anastomosis results in an unacceptable failure rate. Furthermore, sutures encircling the left anterior descending artery should not be used for vessel stabilization as injury of the artery may occur. PMID- 9203602 TI - Less invasive arterial CABG on a beating heart. AB - BACKGROUND: Competitive status of percutaneous transluminal coronary angioplasty and stenting has stimulated an interest in minimally invasive direct coronary artery bypass grafting. METHODS: Between April 1994 and September 1996,156 patients with a mean age of 67 +/- 10 years have undergone minimally invasive direct coronary artery bypass grafting via minithoracotomy, subxiphoid incision, or both with internal mammary artery, right gastroepiploic artery, and radial artery grafting using local coronary occlusion on a beating heart with immobilization of the coronary artery target sites with traction sutures and mechanical regional cardiac wall immobilization platform. RESULTS: Morbidity included wound infection (3), reoperation for bleeding (5), atrial fibrillation (12), central nervous system complication (1), and perioperative myocardial infarction (5). Cardiac-related operative mortality was 1.2% (2/156). Predominantly single grafting was done in 128 patients. Routine angiographic and Doppler echocardiographic flow assessment of anastomotic patency showed an overall patency rate of 92%. In 52 recent consecutive patients in whom the regional cardiac wall mechanical stabilization platform was used, the patency rate of the left internal mammary artery-to-left anterior descending coronary artery graft was improved to 96.2%. With a mean followup of 9.2 +/- 7.4 months, cardiac event-free interval (percutaneous transluminal coronary angioplasty, reoperative coronary artery bypass grafting, or death) in 111 patients was 91%. CONCLUSIONS: Minimally invasive direct coronary artery bypass grafting is safe and effective with good early and midterm clinical results, especially with left internal mammary artery-to-left anterior descending coronary artery grafting via minithoracotomy. Regional cardiac wall immobilization of coronary artery target sites enhances the early graft patency in a predictable manner (96.2%), and this method should be an essential part of all minimally invasive direct coronary artery bypass graft operations with left internal mammary artery-to-left anterior descending artery grafts via minithoracotomy. Further study is required to establish the long-term efficacy of minimally invasive direct coronary artery bypass grafting and the treatment of coronary artery disease. PMID- 9203603 TI - Minimally invasive coronary artery bypass grafting on a beating heart. AB - BACKGROUND: We reviewed our experience with left internal mammary artery (LIMA) to-left anterior descending artery (LAD) anastomosis on a beating heart through a left anterior small thoracotomy. METHODS: This procedure was performed in 343 of 358 scheduled patients; in 15 (4.2%) the LAD was not suitable or was too small. The chest was opened in the fourth (127, 37.0%) or fifth (197, 57.4%) intercostal space, or both (19, 5.6%); the length of the harvested LIMA was 4-15 cm. The LAD was occluded by means of two 4-0 Prolene (Ethicon, Somerville, NJ) sutures, both snared on a small piece of silicone tubing. The anastomosis was performed with two 8-0 Prolene sutures. In the early postoperative period all patients underwent angiography or a doppler flow assessment of the LIMA or both. RESULTS: In 310 patients the LIMA was connected directly to the LAD; to elongate the LIMA, in 30 patients an inferior epigastric artery and in 3 patients a saphenous vein was used. In 2 patients the diagonal branch was also grafted using an inferior epigastric artery from the LIMA. Three patients (0.9%) died during the first 30 days after the operation, and 4 other patients (1.2%) died after the first month. Twenty-five patients (7.3%) were reoperated on because of anastomotic or conduit failure, 18 (5.2%) early and 7 (2.1%) late; one additional patient had a late percutaneous transluminal coronary angioplasty for anastomotic stenosis. At a mean of 9.5 +/- 5.7 months of follow-up, 336 patients (98.0%) were alive, asymptomatic with or without medical treatment, and without cardiac events. COMMENT: Left internal mammary artery-to-LAD anastomosis performed on a beating heart through a left anterior small thoracotomy is a procedure that can be performed with low risk and acceptable midterm results in selected patients. PMID- 9203604 TI - Local immobilization of the left anterior descending artery for minimally invasive coronary bypass grafting. AB - We describe a device for coronary artery stabilization during minimally invasive coronary artery bypass grafting performed without cardiopulmonary bypass via a small (8 to 10 cm) left anterolateral thoracotomy. This device facilitates the anastomosis of the left internal mammary artery to the left anterior descending coronary artery on the beating heart. The device consists of a simple coronary stabilizer mounted on a wound spreader. We have used this device successfully in 35 primary minimally invasive coronary artery bypass grafting operations. PMID- 9203605 TI - Off-bypass coronary bypass grafting via minithoracotomy using mechanical epicardial stabilization. AB - BACKGROUND: Minimally or less invasive surgical coronary revascularization has gained increasing interest along with new techniques and devices designed for easier and safer procedures. Until recently, it appeared questionable whether grafting techniques with avoidance of cardiopulmonary bypass techniques would allow adequate results compared with conventional techniques using cardioplegic arrest. METHODS: Since June 1996, minimally invasive direct coronary artery bypass grafting procedures without cardiopulmonary bypass were intended in 24 patients (19 male, 5 female; age, 60.5 +/- 10.5 years) applying a special system (CardioThoracic Systems, Inc) for internal mammary artery access and epicardial surface stabilization approaching through an anterolateral minithoracotomy. Neither video-assisted preparation nor additional pharmacologic stabilization was applied. Concomitant risk factors and associated comorbidity were frequent. RESULTS: The procedure was completed in 23 patients, grafting the left anterior descending coronary artery (n = 21) or diagonal branches (n = 3, 1 sequential) as scheduled. In 1 case with internal mammary artery dissection, cardiopulmonary bypass and sternotomy became necessary. Simultaneous carotid endarterectomy was performed in 1 patient. There were two episodes of intraoperative ventricular fibrillation; no other major complications occurred. Postoperative evaluation was obtained in 16 patients (15 by angiography, 1 by Doppler echocardiography) so far and revealed adequate graft function and patency. CONCLUSIONS: Using specially designed instruments for internal mammary artery access and epicardial surface stabilization, minimally invasive direct coronary artery bypass grafting procedures via a minithoracotomy avoiding cardiopulmonary bypass techniques may be applied safely and successfully, even in increased risk constellations. PMID- 9203606 TI - Resource utilization for minimally invasive direct and standard coronary artery bypass grafting. AB - BACKGROUND: Minimally invasive direct coronary artery bypass grafting (MIDCABG) has been recently reintroduced into the cardiac surgical armamentarium for selected patients with suitable coronary anatomy. We hypothesized that MIDCABG had the potential for similar immediate results with decreased perioperative morbidity and decreased resource utilization compared with standard coronary artery bypass grafting (CABG). METHODS: From January 1996 to August 1996, 17 MIDCABG patients were compared with 33 patients with left ventricular ejection fraction greater than 0.50 who underwent CABG with standard technique. No significant differences were observed between the two groups for preoperative variables that are known to affect cost and resource utilization. Length of stay in the hospital was 2.5 +/- 0.8 days for MIDCABG and 5.9 +/- 2 days for CABG (p < 0.0001); length of stay in the intensive care unit was 12.3 +/- 3.3 hours for MIDCABG compared to 32.3 +/- 12.6 hours for the CABG group (p < 0.0001). RESULTS: Forty-one percent of MIDCABG patients were extubated in the operating room and 59% were discharged home on the first or second postoperative day versus none in the CABG group (p < 0.0001). Significantly less morbidity was observed in the MIDCABG group compared with CABG. Total ratio of cost-to-charge was $12,885 +/- $1,511 for MIDCABG and $21,260 +/- $5,497 for CABG (p < 0.0001), with an average savings of $8,375. CONCLUSIONS: Minimally invasive CABG is associated with significant reduction of resource utilization and morbidity related to inital hospitalization compared with CABG. PMID- 9203607 TI - Hemodynamic changes during displacement of the beating heart by the Utrecht Octopus method. AB - BACKGROUND: Coronary bypass grafting of posterior circumflex branches requires full displacement of the heart (apex pointing ventrally), which, in the beating heart, results in an arterial pressure drop. We analyzed its origin. METHODS: To facilitate displacement, the Utrecht "Octopus" method was applied in 8 anesthetized beta-blocked pigs and the beating heart was fully retracted. RESULTS: Displacement decreased stroke volume from 75 +/- 17 mL (mean +/- standard deviation) to 43 +/- 13 mL (p < 0.001), a 44% +/- 3% decrease that resulted in a decrease in cardiac output by 32% +/- 5% (mean +/- standard error of the mean; p < 0.001), a decrease in mean arterial pressure by 26% +/- 5% (p < 0.01), and an increase in heart rate by 26% +/- 6% (p < 0.01). Right ventricular end-diastolic pressure increased from 5 +/- 1 to 8 +/- 1 mm Hg (p < 0.01). Twenty degrees head-down tilt normalized cardiac output and mean arterial pressure. Right ventricular end-diastolic pressure increased to 10 +/- 2 mm Hg (p < 0.001) and left ventricular end-diastolic pressure to 11 +/- 3 mm Hg (p < 0.01). CONCLUSIONS: Displacement of the beating heart in the pig induced a 44% drop in stroke volume, which is attributed to biventricular interference with pump function. The Trendelenburg maneuver reestablished the control circulatory status at the expense of augmented right and left ventricular preloads and an increased heart rate. PMID- 9203608 TI - Experimental off-pump grafting of a circumflex branch via sternotomy using a suction device. AB - BACKGROUND: We have shown previously in the pig that coronary artery bypass grafting on the beating heart may be facilitated by local cardiac wall immobilization by a suction device ("Octopus") applied to the anterolateral side of the heart. The purpose of this study was to investigate the feasibility of the method on the posterolateral side. METHODS: In a consecutive series of 8 pigs, after median sternotomy, the posterior wall was taken hold of by the Octopus and subsequently brought up anteriorly and immobilized while hemodynamics were monitored. A posterolateral branch of the circumflex artery was grafted with the left internal mammary artery. After the coronary artery was ligated proximally, the heart was repositioned. At 6 weeks, bypass graft angiography and functional testing (postocclusion hyperemia testing) were performed. After sacrifice, histologic examination of the anastomosis was performed. RESULTS: Dislocation of the heart to expose the distal anastomosis site caused a minor drop in mean arterial blood pressure from 71 +/- 14 (baseline) to 63 +/- 6 mm Hg (dislocated) (not significant) and recovery to 70 +/- 12 mm Hg, 15 minutes after repositioning. Cardiac output decreased from 4.0 +/- 1.0 to 3.2 +/- 0.7 L/min (p = 0.02) and recovered to 4.3 +/- 0.3 L/min. No inotropic drugs were necessary. Anastomosing required 21.5 +/- 6.5 minutes. Baseline graft flow was 8 +/- 3 mL/min and increased threefold to 24 +/- 10 mL/min (p < 0.05) at postocclusive hyperemia testing. At sacrifice after 6 weeks (n = 8), graft flow increased fourfold from 5 +/- 2 to 20 +/- 8 mL/min (p = 0.002) (n = 7). At histologic examination all eight anastomoses were patent without stenosis or mural thrombus. CONCLUSIONS: Off-pump coronary artery bypass grafting of the posterolateral circumflex branches using the Octopus method on the beating pig heart is feasible, with full patency maintained for at least 6 weeks. PMID- 9203609 TI - Instrumentation for minimally invasive internal thoracic artery harvest. AB - The anterior fourth interspace minithoracotomy is our current choice for exposure of the anterior myocardial wall for minimally invasive coronary bypass grafting procedures. This approach provides direct access to the left anterior descending coronary artery for anastomosis, and good exposure of the midsegment of the internal thoracic artery. We describe the use of instrumentation that facilitates the harvest of the left internal thoracic artery under direct vision. The use of this retractor system, which elevates the third and fourth and depresses the second and first ribs, permits better visualization of the internal thoracic artery and allows for proximal internal thoracic artery harvest without rib resection. PMID- 9203610 TI - Video-assisted coronary bypass grafting on the beating heart. AB - BACKGROUND: Concepts to make coronary artery bypass operations less invasive include minimization of access incisions, elimination of cardiopulmonary bypass, and no manipulation of the aorta. A spectrum of minimally invasive coronary bypass procedures now exist that eliminate the median sternotomy (port-access approach), cardiopulmonary bypass ("off-pump bypass"), or both (minimally invasive direct coronary artery bypass procedure). Although all have advantages in decreasing the morbidity of myocardial revascularization, significant limitations exist including surgeon experience, access, exposure, visualization, hemodynamic support, and technique of anastomosis. METHODS: In an attempt to extend the applicability of the current minimally invasive techniques, efforts are being made both to extend the indications for the procedure as well as to modify the technical aspects. Our current experimental protocol involves a ports only approach with three-dimensional video imaging, percutaneous Hemopump circulatory support, Octopus coronary immobilization, and an endoscopically sutured coronary anastomosis. RESULTS: In a series of animal studies we have been able to successfully accomplish a totally endoscopic coronary artery bypass procedure on a beating heart without cardiopulmonary bypass. CONCLUSIONS: Although endoscopic coronary artery bypass without cardiopulmonary bypass is possible, many technical challenges remain. Three-dimensional video imaging, wall motion immobilization and presentation, and an axial-flow device can facilitate the procedure. Future enabling technology including motion robotics and nonvisual imaging systems may ultimately further minimize the invasiveness of surgical myocardial revascularization. PMID- 9203611 TI - Thoracoscopic internal mammary artery harvesting: technical considerations. AB - BACKGROUND: Technical details of thoracoscopic harvesting of the internal mammary artery (IMA) are reported. This procedure allows a complete dissection of the left IMA from its origin at the subclavian artery to the sixth intercostal branches with transection of all collateral branches. METHODS: Between September 1995 and September 1996, thoracoscopic harvesting of the left IMA was performed on 32 patients who had undergone a minimally invasive coronary artery bypass grafting procedure. RESULTS: There were no conversions to a standard approach because of an injury to the graft and no reoperations for bleeding. The mean duration of the IMA harvesting procedure was 58.7 minutes (range, 20 to 130 minutes). CONCLUSIONS: This procedure enlarges the field of minimally invasive coronary artery bypass grafting techniques. The thoracoscopic harvest of the full length of the IMA allows the procedure to more closely replicate the open approach. PMID- 9203612 TI - Thoracoscopic internal mammary artery harvest for MICABG using the Harmonic Scalpel. AB - BACKGROUND: Thoracoscopic internal mammary artery (IMA) harvest is technically demanding, particularly on the left side. We have devised a Harmonic Scalpel (Ethicon Endo-Surgery, Cincinnati, OH) technique to facilitate this procedure, and describe our clinical experience here. METHODS: The Harmonic Scalpel functions with ultrasonic energy, producing less smoke and lower heat than regular electrocautery. A total of 27 (22 left and 5 right) pedicles of the IMA in 23 patients were harvested from the upper margin of the first rib or higher to the lower margin of the fifth rib thoracoscopically using the Harmonic Scalpel with a hook blade. RESULTS: In each case, the IMA harvest was completed thoracoscopically with only the Harmonic Scalpel, decreasing instrument transfers. Each vascular branch was coagulated without charring and was transected with excellent hemostasis. Smokeless views were provided. In the first 17 harvests, Doppler studies 3 months after the procedures demonstrated patent IMAs to the coronary circulation. CONCLUSIONS: The Harmonic Scalpel facilitates thoracoscopic IMA harvest and is expected to minimize hyperthermic damage of the IMA. PMID- 9203613 TI - VATS for complete dissection of LIMA in minimally invasive coronary artery bypass grafting. AB - BACKGROUND: The aim of this work is to report our initial experience with minimally invasive coronary artery bypass grafting, using video-assisted thoracic surgery (VATS) to facilitate the operation and provide complete dissection of the left internal mammary artery (LIMA). METHODS: Of 44 scheduled patients, 43 patients, 30 (69.8%) male, ranging in age from 31 to 83 years (60.8 +/- 12.0 years), with a severe lesion in the anterior descending artery, were operated upon. An 8-cm left anterior minithoracotomy was performed at the fourth intercostal space. Through this incision the optical device for video-assisted thoracic surgery as well as the surgical instruments were placed to provide complete LIMA dissection. This permits dissection until the subclavian region, allowing for anastomosis without tension or distortion. Bypass circulation was not used, and the cardiac rate was decreased with the use of intravenous beta blockers. For LIMA-to-anterior descending artery anastomosis, proximal and distal tourniquets were used and 1.5 mg/kg of heparin was administered intravenously. RESULTS: Video-assisted thoracic surgery provided a complete dissection of LIMA. The 43 patients presented satisfactory postoperative progress, being released from the hospital between 2 and 12 days after their operation, with a mean of 4 days. The patients have remained asymptomatic during a period that ranged from 1 to 13 months (6.3 +/- 3.5 months). During the follow-up, there was one death as a result of stroke and pneumonia 2 months after the release from the hospital. CONCLUSIONS: The use of video-assisted thoracic surgery through thoracotomy allows the LIMA dissection without the necessity of other incisions. The procedure also permitted more ample dissection of LIMA when compared with minithoracotomy without video-assisted thoracic surgery. PMID- 9203614 TI - Mammary-coronary artery anastomosis without cardiopulmonary bypass through a minithoracotomy. AB - BACKGROUND: Coronary artery bypass grafting has been based on cardiopulmonary bypass, myocardial protection, and the median sternotomy. The recent concept of minimally invasive coronary artery bypass grafting in selected patients has dramatically affected surgical management of coronary artery disease. Coronary artery bypass grafting of anterior coronary arteries with in situ internal mammary artery through a limited anterior thoracotomy is a procedure that is gaining acceptance. METHODS: Fifty-one patients were operated on by minithoracotomy and direct coronary artery bypass grafting without cardiopulmonary bypass. Left internal mammary artery-to-left anterior descending coronary artery anastomosis was done in 50 patients, and in 1 patient, left internal mammary artery-to-left anterior descending artery and right internal mammary artery-to-right coronary artery anastomoses were constructed through bilateral minithoracotomies. Left anterior minithoracotomy through the fourth intercostal space and right anterior minithoracotomy through the fifth intercostal space were used for left internal mammary artery and right internal mammary artery dissection, respectively. With this approach, a 4- to 6-cm length of mammary artery was easily dissected. Mammary-to-coronary anastomosis was performed on a beating heart without cardiopulmonary bypass through window pericardiotomy. RESULTS: Twenty-five patients were extubated in the operating room and 26 in the intensive care unit 4 to 6 hours after operation. None of these patients required blood transfusion or inotropic support. Postoperative predischarge angiography in 42 patients revealed adequate mammary-to-coronary flow in 40 patients. Doppler flow studies were also in accordance with angiographic findings. Forty-five patients are in our regular follow-up (mean follow-up, 6.23 +/- 1.34 months); 44 of them are in functional class I. CONCLUSION: In our experience minithoracotomy is a safe, simple, and minimally invasive procedure. Favorable cost/benefit ratio has been achieved owing to no early or late mortality and minimal early morbidity. Postoperative angiography and Doppler flow study revealed excellent predictive long-term results. PMID- 9203615 TI - Minimally invasive harvest of the saphenous vein for coronary artery bypass grafting. AB - BACKGROUND: Preparation of the great saphenous vein for coronary artery bypass grafts is traditionally performed through one or many long cutaneous incisions. We describe the dissection of the great saphenous vein through small cutaneous incisions using the Mini Harvest System. METHODS: The device is composed of a retractor coupled to a light source. Introduced under the skin, above the anterior plane of the vein, it allows an easy preparation of the vein under direct vision. The entire vein can be dissected from the ankle to the groin through sequential cutaneous incisions along the leg. A prospective, randomized trial was performed to compare the minimally invasive vein harvest technique (group 1, n = 15) versus the standard method (group 2, n = 15). RESULTS: In addition to an internal mammary artery graft, 28 venous coronary bypass grafts were performed in group 1 (mean, 1.9 +/- 1.0) and 32 in group 2 (mean, 2.1 +/- 1.0). The mean cutaneous incision length divided by the mean length of the harvested vein was 10.8 cm/32.3 cm = 33% for group 1 and 37.6 cm/33.2 cm = 113% in group 2 (p < 0.001). Wounds were examined daily and a final assessment was performed on day 7. Better cicatrization, less hematoma and edema, and less pain were observed in group 1. CONCLUSIONS: Minimally invasive harvest of the great saphenous vein offers many advantages over the traditional harvest method. Besides the aesthetic aspect, healing is better and postoperative discomfort is reduced. PMID- 9203616 TI - Review of facilitated approaches to vascular anastomosis surgery. AB - BACKGROUND: There is an increasing demand for an easier, quicker, less damaging, but reliable procedure to create a vascular anastomosis. This demand is not new but is revitalized by the movement of vascular procedures in various specialties, including cardiac surgery, toward minimally invasive procedures. This article reviews the most important representatives of devices or methods that have been developed in the last two centuries. METHODS: A thorough literature search was performed. The outcome is presented and discussed in four parts: (1) stapling and clipping devices, (2) coupling devices, (3) glues, and (4) laser welding. RESULTS: Stapling devices have not become the standard fashion to create an anastomosis because they are too complicated to use. In selected cases clips have potential in vascular surgery. There is a ring-pin coupling system available that is easy to use and especially suitable for creating an end-to-end anastomosis. The ideal glue is yet to be developed, and the currently available laser welding techniques have to become refined. CONCLUSIONS: It is anticipated that the future lies in hybrid techniques that combine sutures or clips with glues or laser welding techniques. PMID- 9203617 TI - Scanning electron microscopic analysis of the stapled microvascular anastomosis in the rabbit. AB - BACKGROUND: Despite the high percentages of experimental and clinical patency rates achieved using so-called mechanical anastomotic devices (Unilink; 3M, St. Paul, MN; vascular staples) they remain little known and occasionally used. METHODS: The VCS Auto Suture microstapler technique for microvascular anastomosis was tested experimentally and compared with the conventional "gold standard" 10/0 end-to-end microvascular technique. Thirty carotid arteries on one side of 30 rabbits were stapled using nonpenetrating 0.9-mm (small) VCS Auto Suture microclips, and the other 30 carotid arteries on the other side were sutured in a conventional way with 10/0 monofilament nylon. A 100% patency rate was achieved on both sides. Biopsy was performed in five groups of rabbits at different time intervals postoperatively, and the specimens were examined under scanning electron microscopy. RESULTS: All 60 anastomoses were patent. Histomorphologic examination of the anastomotic site revealed no major differences between sutured and stapled groups. CONCLUSIONS: Stapled microvascular anastomosis technique is fast and reliable. PMID- 9203618 TI - Nonpenetrating clips for coronary anastomosis. AB - BACKGROUND: A nonsuture clip technique (nonpenetrating titanium clips applied to everted tissue edges at high compressive forces) was used to perform coronary anastomoses in a clinical setting. METHODS: Clipped coronary anastomoses were performed in 10 patients. The anastomoses incorporated the left internal mammary artery to the left anterior descending artery (n = 1) and the saphenous vein to the right coronary artery (n = 5), the posterior descending artery (n = 2), the diagonal artery (n = 2), and one vein-to-vein proximal anastomosis (n = 1). RESULTS: The mean duration for completion of the anastomoses was 15 minutes (range, 7 to 20 minutes). This time was reduced from 20 minutes at the beginning of the clinical experience to 7 minutes for the last 3 patients. No technical complication was related to clip application and all patients had uneventful outcomes. Three anastomoses studied by coronary angiography were patent without stenosis. CONCLUSION: The clipped anastomotic technique has a rapid learning curve, the same safety as suture methods, and the potential for facilitating endoscopic vascular reconstructions. PMID- 9203619 TI - Nonocclusive excimer laser-assisted end-to-side anastomosis. AB - BACKGROUND: High-flow extraintracranial bypass operation on the brain is a risky procedure because of the temporary occlusion of the intracranial portion of the internal carotid artery. We therefore developed a nonocclusive anastomosis technique in the experimental animal laboratory in 100 chronic and acute experiments in rabbits. METHODS: In 40 patients we interposed a venous transplant between the external carotid artery or one of its branches and the intracranial portion of the internal carotid artery. During the construction of the distal anastomosis the recipient artery was not occluded. The donor vessel was stitched to the exterior of the recipient vessel and an Excimer laser catheter (Medolas GmbH, Amberg, Germany) was introduced by way of an artificial side branch. The tip of the laser catheter created a hole in the wall of the recipient artery just inside the anastomosis. The cut-out full-thickness portion of recipient vessel wall remained attached to the tip of the laser catheter by way of high vacuum suction and was removed together with the laser catheter. The artificial side branch was occluded with a hemostatic clip. No interruption of blood flow in the recipient artery was induced during the making of the anastomosis. RESULTS: The procedure was well tolerated by the patients and a high patency rate was observed. CONCLUSIONS: The nonocclusive Excimer laser-assisted anastomosis technique is safe and yields a high long-term patency rate in neurosurgical patients. It cannot be excluded that there are indications for this method in coronary bypass surgery. PMID- 9203620 TI - Novel subtype-selective nonpeptide bradykinin receptor antagonists FR167344 and FR173657. AB - We describe the receptor binding and antagonistic properties of two novel nonpeptide antagonists, FR167344 (3-bromo-8-[2,6-dichloro-3-[N-[(E)-4-(N,N dimethylcarbamoyl)cinnamido acetyl]-N-methylamino]benzyloxy]-2-methylimidazo[1,2 a]pyridine hydrochloride) and FR173657 (8-[3-[N-[(E)-3-(6-acetamidopyridin-3 yl)acryloylglycyl]-N-m ethylamino]-2,6-dichlorobenzyloxy]-2-methylquinoline), for the human bradykinin receptor subtypes (B1 and B2). In competitive experiments using membranes prepared from Chinese hamster ovary cells expressing the bradykinin receptor subtypes, FR167344 and FR173657 showed a high affinity binding to the B2 receptor with IC50 values of 65 and 8.9 nM, respectively, and no binding affinity for the B1 receptor. FR167344 and FR173657 inhibited the B2 receptor-mediated phosphatidylinositol (PI) hydrolysis and produced a concentration-dependent rightward shift in the dose-response curve to bradykinin. This shift was accompanied by a progressive reduction of maximal response. Estimated pA2 values for the antagonism of bradykinin-induced PI hydrolysis by FR167344 and FR173657 were 8.0 and 9.0, respectively. FR167344 and FR173657 showed no stimulatory effects on PI hydrolysis. Therefore, FR167344 and FR173657 are potent, highly selective, and insurmountable antagonists for the human bradykinin B2 receptor. PMID- 9203621 TI - Internal trafficking and surface mobility of a functionally intact beta2 adrenergic receptor-green fluorescent protein conjugate. AB - The beta2-adrenergic receptor (beta2AR) is prototypic of the large family of G protein-coupled receptors (GPCRs) whose desensitization and resensitization are regulated by intracellular kinases, arrestin proteins, phosphatases, and ill defined components of the cellular endocytic machinery. The study of beta2AR signal transduction and behavior in living cells is technically difficult because of the relatively low cellular expression of the receptor and a lack of useful biological reagents. Availability of a functional beta2AR tagged with the highly sensitive Green Fluorescent Protein (GFP) could allow measurements of the various properties of the beta2AR. We demonstrate that a fully functional beta2AR/GFP can be engineered. In mammalian cells, beta2AR/S65T/GFP demonstrates strong, diffuse plasma membrane fluorescence when observed with 480 nm excitation. The fluorescent receptor binds agonist and antagonist, stimulates adenylyl cyclase, undergoes phosphorylation, and is internalized in a manner indistinguishable from wild-type receptor. We then show that its internal trafficking and surface mobility can be determined by measuring only the endogenous fluorescence of the conjugate. beta2AR/S65T/GFP was found to be localized on endosomal membranes in living cells within minutes of agonist treatment, and within 15 min it is observed in more complicated structures formed from fusion of multiple endosomes. Finally, its free diffusion (diffusion coefficient, 4.0-12 x 10(-9) cm2/sec) was assessed on living cells using photobleaching recovery measurements. This approach and the fidelity of the biochemical properties of the beta2AR/S65T/GFP demonstrate that real-time optical measurements of beta2AR (as well as other GPCR) interactions and dynamics on living cells are feasible. PMID- 9203622 TI - Roles of cholecystokinin receptor phosphorylation in agonist-stimulated desensitization of pancreatic acinar cells and receptor-bearing Chinese hamster ovary cholecystokinin receptor cells. AB - Receptor phosphorylation has been implicated in desensitization responses to some agonist ligands, in which receptors may become uncoupled from G proteins and move into cellular compartments inaccessible to hydrophilic ligands. Understanding of the linkage between these processes, however, has come largely from recombinant receptor-bearing cell systems with consensus sites of kinase action mutagenized. We recently established methodology permitting direct assessment of sites of phosphorylation of the cholecystokinin receptor (CCKR) in its native milieu in the pancreatic acinar cell and in a Chinese hamster ovary (CHO)-CCKR cell line (1, 2). Although CCK binding leads to phosphorylation of serine residues within the third intracellular loop of the receptor in both cell types, there are clear differences in the time course of phosphorylation, in the balance of action of kinases and a receptor phosphatase, and in a few of the distinct sites phosphorylated. In this work, we have directly assessed the inositol 1,4,5 triphosphate responses to CCK and desensitization of these responses in both cells. CHO cell lines expressing receptor mutants with protein kinase C consensus sites modified were also studied. CCK-stimulated inositol 1,4,5-triphosphate responses in both cells expressing wild-type receptors were rapidly and completely desensitized, associated with the onset of receptor phosphorylation. However, despite maintenance of the phosphorylated state of the receptor in the CHO-CCKR cell and its dephosphorylation returning the receptor to its basal state in the acinar cell, desensitization continued to be present in both. Mutagenesis of Ser260 and Ser264 to alanines individually reduced receptor phosphorylation by approximately 50%, whereas the dual mutant completely eliminated agonist stimulated phosphorylation. Because other sites of phosphorylation were still intact in this construct, this raises the possibility of hierarchical phosphorylation with these two sites key in making other sites accessible to kinases. Constructs modifying Ser264 delayed the onset of desensitization, whereas all constructs proceeded to achieve complete desensitization by 10 min. Receptor internalization occurred independent of its phosphorylation state in the CHO cell lines, explaining the desensitization observed. In the acinar cell in which the receptor remains on the cell surface after agonist occupation, we postulate that receptor insulation achieves similar uncoupling from G protein association as is achieved by receptor phosphorylation early after agonist occupation. PMID- 9203623 TI - Genistein-induced apoptosis of prostate cancer cells is preceded by a specific decrease in focal adhesion kinase activity. AB - Genistein (5,7,4'-trihydroxyisoflavone), an isoflavinoid found in soy beans, has been identified as potentially causal for the low incidence of metastatic prostate cancer (PCa) in certain countries. Although genistein-induced PCa cell adhesion has been identified as a possible causative mechanism, direct growth inhibition by genistein has been reported and also could be causal. If in vivo growth inhibition was significant, then growth inhibition should occur at concentrations attained with dietary consumption, the mechanism of growth inhibition should be relevant to PCa, and genistein (a broad-spectrum in vitro protein-tyrosine kinase inhibitor) should have relatively specific kinase inhibitory effects in vivo. These considerations were investigated by measuring growth inhibitory activity in a variety of PCa cell lines. Growth inhibitory effects were shown not to occur with concentrations below the low micromolar range (i.e., 3 logs above that attained in serum). In-depth mechanistic studies with the PC3-M metastatic variant cell line demonstrated that growth inhibition was independent of genistein's estrogenic effects. Genistein was shown to decrease the viability of nonadherent cells, suggesting a lack of dependence on cell adhesion for growth inhibition. However, important molecular and kinetic differences between genistein's effects on growth in adherent versus nonadherent cells were identified. Specific suppression of focal adhesion kinase activity (without global decreases in phosphotyrosine) was shown to precede induction of apoptosis, which was responsible for growth inhibition in adherent cells. These findings do not support an in vivo growth inhibitory role by genistein consumed in quantities associated with a soy-based diet. They do, however, identify genistein as a potential therapeutic agent for PCa and as a tool with which to study the control of apoptosis in PCa. PMID- 9203624 TI - Beta-adrenoceptor activation-induced placental prorenin secretion is mediated by increased renin messenger RNA and protein synthesis. AB - Activation of beta-adrenoceptors has been shown to promote renin secretion in both human kidney and placenta. In kidney, the enhanced secretion is immediately observed, and mobilization of renin in the storage granules accounts for such a rapid response. In contrast, the enhanced secretion in placenta is delayed for 6 12 hr after receptor activation and consists almost entirely of the renin precursor prorenin. It is hypothesized that newly synthesized rather than stored enzyme is responsible for the enhanced secretion in human placenta. To test this hypothesis, placental explants were cultured in the presence or absence of the protein synthesis inhibitor cycloheximide, and prorenin concentrations in the tissue and medium were measured. Dobutamine and terbutaline, beta1- and beta2 adrenoceptor agonists, evoked 17- and 5-fold increases in secretion, respectively. Tissue content of prorenin in response to the treatment was increased by a similar magnitude, yet values were consistently <10% of medium concentrations. The increases in prorenin concentrations in both medium and tissue, however, were markedly attenuated by cycloheximide, suggesting that prorenin synthesis in response to beta-adrenoceptor activation is required. Reverse transcription coupled with polymerase chain reaction revealed that renin mRNA levels were increased by 3-8-fold and occurred before increases in tissue and medium prorenin, indicating that increased renin mRNA levels are responsible for the increased synthesis of prorenin. Explants cultured in the presence of actinomycin D, an inhibitor of transcription, did not show the agonist-induced prorenin mRNA levels or enhancement of its secretion. The peak levels of renin mRNA were reached after 6 hr of incubation, were sustained at similar levels after 24 hr, and were not affected by cycloheximide. These findings are consistent with the notion that enhancement of renin mRNA and de novo protein synthesis are required for prorenin secretion induced by activation of placental beta-adrenoceptors. PMID- 9203625 TI - Antisense oligonucleotides targeting human protein kinase C-alpha inhibit phorbol ester-induced reduction of bradykinin-evoked calcium mobilization in A549 cells. AB - Regulation of the bradykinin-evoked increase in intracellular Ca2+ concentration by protein kinase C (PKC)-alpha was investigated in A549 human lung carcinoma cells. Bradykinin, a potent and selective kinin B2 receptor agonist, induces calcium mobilization in a concentration-dependent fashion in this cell line. 12-O Tetradecanoylphorbol-13-acetate (TPA), a potent activator of PKC, is known to reduce the amplitude of agonist-induced calcium mobilization in various cell lines. Because PKC-alpha is a major PKC isozyme in A549 cells, we investigated whether this isozyme plays a role in this process. A 20-mer phosphorothioate oligonucleotide targeting the 3'-untranslated region of the human PKC-alpha mRNA, which contains 2'-methoxyethyl modifications incorporated into the 5' and 3' segments of the oligonucleotide, was used to assess the putative role of PKC alpha in the receptor regulation. ISIS 9606 reduced PKC-alpha mRNA for > or = 72 hr after the initial treatment and the reduction was concentration dependent, whereas the mismatch control, ISIS 13009, had no effect. Concentrations of ISIS 9606 of 150 nM specifically reduced the level of immunoreactive PKC-alpha protein by 66.3 +/- 2.5% at 72 hr after treatment, without an effect on immunoreactive PKC-delta protein. This reduction in PKC-alpha was sufficient to inhibit the reduction of bradykinin-induced calcium mobilization by TPA. This finding is corroborated by the use of staurosporine, a nonselective PKC inhibitor, that prevented the effect of TPA. These results suggest that PKC-alpha is involved in kinin B2 receptor regulation by phorbol esters in A549 cells. PMID- 9203626 TI - A new linear V1A vasopressin antagonist and its use in characterizing receptor/G protein interactions. AB - We characterized a new iodinated, high affinity, linear V1a vasopressin antagonist, phenylacetylD-Tyr(Et)Phe-Gln-Asn-Lys-Pro-Arg-Tyr-NH2. The antagonist bound specifically to the V1a vasopressin receptor in crude rat liver membranes with an apparent Kd value of 0.168 nM. This affinity is approximately 1 order of magnitude greater than that of the natural agonist, vasopressin. The inhibitory activity of the antagonist can be demonstrated by its inability to elicit activation and uncoupling of G proteins from the receptor. Thus, after occupancy of receptor sites in rat liver membranes with labeled antagonist and detergent solubilization, the labeled receptor (approximately 60 kDa) was eluted as a stable 400-kDa complex on size-exclusion chromatography. In contrast, when the receptor sites were occupied by the agonist [3H]vasopressin, the receptor eluted as a 60-kDa peak. Coincubation of membranes with iodinated antagonist and an excess of unlabeled vasopressin caused both reduced antagonist binding and a complete shift from the 400-kDa to the 60-kDa peak. The addition of vasopressin to unliganded 400-kDa fractions resulted in a 75% increase in [35S]guanosine-5'-O (3-thio)triphosphate binding activity, indicating that the 400-kDa fraction contains complexes between the V1a receptor and G proteins. The vasopressin elicited increase was inhibited by antagonist. Using specific antibodies and immunoadsorption to protein A/Sepharose columns, we found that G protein isotypes G(alpha q/11), G(alpha i3), and G(alpha s), and effector enzymes PLC-beta1, PLC gamma2 and PLA-2 were associated with the antagonist-labeled receptor in the 400 kDa fraction. Because the 400-kDa complex was found in the absence of ligand, the V1a receptor and the appropriate G proteins and effector enzymes are likely preassociated with each other and do not aggregate after antagonist addition. The association of V1a receptor with the different specific G proteins and effector enzymes is consistent with the multiple actions of vasopressin on liver cells. Antibodies directed against a portion of the carboxyl-terminal domain of the V1a receptor interacted with 60-kDa antagonist-occupied receptor but not with receptor in the 400-kDa complex. These results suggest that the carboxyl-terminal region of the receptor is sterically hindered when coupled to G proteins. The iodinated linear vasopressin antagonist therefore allows stable receptor/G protein complexes and can be an important tool (along with the antisera) for use in the study of factors that control V1a receptor/G protein coupling. PMID- 9203627 TI - Enhancement of radiation-inducible hepatic glutathione-S-transferases Ya, Yb1, Yb2, Yc1, and Yc2 gene expression by oltipraz: possible role in radioprotection. AB - Previous studies have shown that radiation in combination with oltipraz enhances hepatic microsomal epoxide hydrolase expression. The effects of gamma-ray radiation exposure in combination with oltipraz on the expression of hepatic glutathione-S-transferase (GST) subunits Ya, Yb1, Yb2, Yc1, and Yc2 were examined in the rat. Northern RNA blot analyses revealed that GST mRNA levels were altered in response to daily 3- or 0.5-Gy doses of radiation. The hepatic GST mRNA levels were transiently decreased at 3 and 8 hr after a single 3-Gy dose of radiation. The GST Ya, Yb1, Yb2, Yc1, and Yc2 mRNA levels were increased by 2-4-fold at 15 and 24 hr after irradiation with 3 Gy, followed by return to the levels of untreated rats at 48 hr after treatment. The treatment of animals with oltipraz alone resulted in dose-related increases in the GST Ya, Yb1, Yc1, and Yc2 mRNA levels, whereas Yb2 mRNA levels were, minimally increased. Although a single dose of oltipraz (30 mg/kg orally) caused a minimal 2-fold elevation in the hepatic GST Ya mRNA level, exposure of animals to both oltipraz and 3-Gy radiation resulted in a 4-fold relative increase in GST Ya mRNA level, indicating that the Ya mRNA expression was additively enhanced by the combination treatment. The Yb1/2 and Yc1/2 mRNA expressions were also enhanced by oltipraz in combination with radiation. Multiple exposure of rats to daily 0.5-Gy radiation caused time related increases in GST gene expression. The greatest enhancement in GST expression was observed at 24 hr after a single 0.5-Gy dose of radiation in conjunction with oltipraz (e.g., a 9-fold relative increase in GST Ya), whereas the relative additive increases in GST mRNA were less pronounced at day 3 or 5 after treatment. These increases in the GST mRNA levels were consistent with those in the immunochemically detectable GST protein levels. Histopathological examinations revealed that exposure of rats to radiation (0.5 Gy/day for 3-5 days) caused mild-to-moderate hepatocyte degeneration with sinusoidal congestion, whereas oltipraz (30 mg/kg/day for 3 days) was effective in blocking the radiation-induced liver injury. The enhanced expression of these GST isoforms by oltipraz may be associated in part with its hepatoprotective effect against the injury caused by ionizing radiation. PMID- 9203628 TI - The role of the aspartate-arginine-tyrosine triad in the m1 muscarinic receptor: mutations of aspartate 122 and tyrosine 124 decrease receptor expression but do not abolish signaling. AB - An Asp-Arg-Tyr triad occurs in a majority of rhodopsin-like G protein-coupled receptors. The fully conserved Arg is critical for G protein activation, but the function of the flanking residues is not well understood. We expressed in COS-7 cells m1 muscarinic receptors that were mutated at Asp122 and Tyr124. Most mutations at either position strongly attenuated or prevented the expression of binding sites for the antagonist [3H]N-methylscopolamine. However, sites that were expressed displayed unaltered affinity for the antagonist. Receptor protein, visualized with a carboxyl-terminally directed antibody, was reduced but never completely abolished. The effects of these mutations were partially reversed by the deletion of 129 amino acids from the third intracellular loop of the receptor. In several cases, comparison of immunocytochemistry with binding measurements suggested the presence of substantial amounts of inactive, presumably misfolded, receptor protein. Some of the variants that bound [3H]N methylscopolamine underwent small changes in their affinities for acetylcholine. All retained nearly normal abilities to mediate an acetylcholine-induced phosphoinositide response. We propose that Asp122 and Tyr124 make intramolecular contacts whose integrity is important for efficient receptor folding but that they do not participate directly in signaling. The role of these residues is completely distinct from that of Arg123, whose mutation abolishes signaling without diminishing receptor expression. PMID- 9203629 TI - Selective activation of rolipram-sensitive, cAMP-specific phosphodiesterase isoforms by phosphatidic acid. AB - In rat thymic lymphocytes, accumulation of phosphatidic acid (PA) occurs at the same time as decrease in cAMP levels and activation of a cAMP-specific phosphodiesterase (PDE) [type 4, EC 3.1.4.17 (PDE4)]. We investigated the nature of the PDE activated by PA and the mechanism of activation by using recombinant cAMP-specific PDE4 isoforms derived from three different genes (PDE4A, PDE4B, and PDE4D). The "long" variants expressed from each gene (PDE4A5, PDE4B1, and PDE4D3) were activated by PA, whereas the "short" variants (PDE4A1, PDE4B2, PDE4D1, and PDE4D2) were not. Phosphatidylserine was an activator that was as effective as PA, whereas phosphatidylcholine was ineffective, indicating that activation was restricted to anionic phospholipids. PA caused an increase in the Vmax value of PDE4D3 without affecting the Km value of the enzyme for the cAMP substrate. PA also caused a change in the Mg2+ requirement for hydrolysis. Half-maximal stimulation of the PDE was obtained with approximately 10 microg/ml PA. Although protein kinase A-mediated phosphorylation of PDE4D3 produces effects similar to those elicited by PA, the mechanism of PA-induced activation was not found to involve a phosphorylation. Instead, several observations suggest that PA may directly interact with the enzyme. The stimulation of cAMP PDEs by PA and other acidic phospholipids may be a mechanism by which growth factors and hormones modulate the cAMP-dependent signal transduction pathway during cell stimulation. PMID- 9203630 TI - Structural and functional characterization of the human alpha3 nicotinic subunit gene promoter. AB - We describe the structural and functional features of the human alpha3 nicotinic receptor subunit promoter. A 0.35-kb region immediately upstream of the start codon was identified that when transfected in human neuroblastoma cells was able to drive the expression of the luciferase reporter gene with a strength comparable to that of the well-characterized simian virus 40 promoter/enhancer. This region displayed the features of a multistart-site, GC-rich, TATA-less, and CAAT-less promoter, containing many overlapping Sp1 and AP-2 putative binding sites. Further dissections of the 0.35-kb fragment revealed that its 3' region, specifying the 5' UT of the mRNA, plays a relevant positive effect in determining the strength of the promoter. This region contains putative cis-acting elements for AP-2, nuclear factor-kappaB, and the recently described multiple-start site element downstream-1. By mutation analysis, we showed that these sites are functional and when combined increase the promoter activity by 4-fold. The 0.35 kb promoter was found to be under the negative control of upstream sequences that include a modern Alu repeat. The alpha3 Alu repeat works as a composite region, containing both positive and negative elements that control the activity of the downstream promoter. Finally, we investigated the tissue-specific activity of the human alpha3 gene 5' regulatory sequences, showing that they are able to drive the expression of the reporter gene preferentially in neuronal cells. PMID- 9203631 TI - 1,5-benzothiazepine binding domain is located on the extracellular side of the cardiac L-type Ca2+ channel. AB - To determine whether 1,5-benzothiazepine Ca2+ channel blocker approaches its binding domain within the cardiac L-type Ca2+ channel from inside or outside of the membrane, we tested the effects of a novel potent 1,5-benzothiazepine derivative (DTZ323) and its quaternary ammonium derivative (DTZ417) on guinea pig ventricular myocytes by using the whole-cell patch-clamp technique. The extracellular application of DTZ417 suppressed the L-type Ca2+ channel currents (I[Ca(L)]) with an IC50 value of 1.2 +/- 0.02 microM, which was close to the IC50 value of diltiazem (0.63 +/- 0.01 microM). The suppression of I[Ca(L)] by DTZ417 was voltage and use dependent but lacked tonic block, which allowed us to investigate the onset of the effect on I[Ca(L)] by changing the holding potential (HP) from -90 to -50 mV in the presence of DTZ417. DTZ417 did not have significant effects on I[Ca(L)] at an HP of -90 mV. At -50 mV, DTZ417 (50 microM) applied from the extracellular side completely suppressed I[Ca(L)], whereas it had no effect from the intracellular side. DTZ323 (1 microM) also inhibited I[Ca(L)] only from the extracellular side, without any effects by the intracellular application of < or = 10 microM. However, a quaternary phenylalkylamine derivative, D890 (0.1 mM), acted only from the intracellular side. These results suggest that in contrast to the phenylalkylamine binding site, in cardiac myocytes the 1,5-benzothiazepine binding site is accessible from the extracellular side of the L-type Ca2+ channel. PMID- 9203632 TI - Correlation between the formation of cleavable complex with topoisomerase I and growth-inhibitory activity for saintopin-type antibiotics. AB - New saintopin-type antibiotics (e.g., saintopin, saintopin E, UCE1022, UCE6) with a naphthacene-dione structure have been discovered through our mechanistically oriented screening using purified mammalian DNA topoisomerases. Saintopin is a dual inducer of topoisomerase I- and topoisomerase II-mediated DNA cleavages in a cell-free system using purified enzymes, whereas others induced topoisomerase I- but not topoisomerase II-mediated DNA cleavage. The order of topoisomerase I mediated DNA cleavage activity at lower concentrations (<1 microM) was UCE6 > saintopin > saintopin E > UCE1022. The DNA cleavage-intensity patterns induced by these antibiotics with topoisomerase I were identical, indicating that saintopin type antibiotics have a similar DNA sequence selectivity in stabilization of the cleavable complex with topoisomerase I. Increases in protein/DNA complexes were observed in saintopin-type antibiotic-treated HeLa S3 cells using the potassium/sodium dodecyl sulfate precipitation method. Brief heating of these drugs-treated cells at 65 degrees for 10 min resulted in a rapid reduction in the number of protein/DNA complexes. Immunoblot analysis using antibody against human topoisomerase I or II revealed that the protein linked to DNA in saintopin-type antibiotic-treated cells is most likely topoisomerase I. These results suggest that saintopin-type antibiotics interfere with topoisomerase I in cells by trapping reversible topoisomerase I/DNA cleavable complexes. The formation of topoisomerase I/DNA complexes by saintopin-type antibiotics correlates well with their growth-inhibitory activities, suggesting that topoisomerase I can be the principal target of these antibiotics. PMID- 9203633 TI - Diinosine polyphosphates, a group of dinucleotides with antagonistic effects on diadenosine polyphosphate receptor. AB - A new family of dinucleotide derivatives, diinosine polyphosphates, has been synthesized through the use of the enzyme 5' adenylic acid deaminase from Aspergillus sp., starting from the corresponding diadenosine polyphosphates. Functional studies were performed on rat brain synaptic terminals in which a dinucleotide receptor has been described that is specific for adenine dinucleotides. The results demonstrated that diinosine polyphosphates did not behave as agonists on the diadenosine polyphosphate receptor (also know as P4 purinoceptor), but they were very efficient as antagonists in abolishing the Ca2+ responses elicited by diadenosine pentaphosphate. The IC50 values for diinosine triphosphate, diinosine tetraphosphate, and diinosine pentaphosphate were 4.90 +/ 0.10 microM, 8.33 +/- 0.22 microM, and 4.23 +/- 0.12 nM, respectively. The diinosine polyphosphates also antagonized the ATP receptors present in synaptic terminals, showing IC50 values of 100.08 +/- 5.72 microM for diinosine triphosphate, 29.51 +/- 1.40 microM for diinosine tetraphosphate and 27.75 +/- 1.65 microM for diinosine pentaphosphate. The antagonistic ability of these diinosine nucleotides was studied in comparison with other P1 and P2 purinoceptor antagonists, such as suramin, pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid, and 8-cyclopentyl-1,3-dipropylxanthine. These purinergic antagonists did not inhibit the response of the P4 purinoceptor; only the diinosine polyphosphates were able to act as antagonists on the dinucleotide receptor. Suramin and pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid attenuated the responses elicited by ATP, as did the diinosine polyphosphate compounds. The most antagonistic diinosine polyphosphate for the dinucleotide and ATP receptors was diinosine pentaphosphate, which was 6000 times more selective for the P4 purinoceptor than it was for the ATP receptor. PMID- 9203634 TI - Cell-specific, promoter-dependent mineralocorticoid agonist activity of spironolactone. AB - The agonist activity of the antimineralocorticoid spironolactone was evaluated in various cell lines through the use of transfection experiments. The target promoters were derived from the deltaMTV promoter in which one or several glucocorticoid-responsive elements (GRE) were inserted in tandem. Spironolactone at 100 nM activated by 6-fold the GRE/deltaMTV promoter in the human hepatoma HepG2 cell line and only partially prevented the 10-fold activation of this promoter by 0.1 nM aldosterone. Both effects were completely dependent on the cotransfection of an expression vector for the mineralocorticoid receptor. The half-maximal agonist effect of spironolactone was similar to its half-maximal antagonist effect (approximately 10 nM). For the GRE-2/deltaMTV, GRE-4/deltaMTV, and wild-type MMTV promoters, the activation by aldosterone was much more potent (70-, 100-, and 110-fold, respectively), whereas spironolactone elicited a 10-, 24-, and 25-fold activation, respectively. Thus, the effect of both compounds and the relative efficiency of spironolactone, compared with that of aldosterone, were dependent on the number of GREs present in the regulatory region of the promoter. The agonist effect of spironolactone was cell specific. Indeed, although spironolactone agonist activity was observed in H5 kidney tubule cells, none could be detected at concentrations of < or = 1 microM in the CV1 monkey fibroblast cells. In contrast, the antagonist effect was observed in all cells. Furthermore, other antimineralocorticoids, such as RU 26752 and progesterone, also displayed mineralocorticoid receptor-dependent agonist activity in the HepG2 cells. The antiprogesterone RU 486 and the antiandrogen cyproterone acetate were ineffective at < or = 1 microM. In conclusion, we show that under certain experimental conditions, several antimineralocorticoids display significant agonist activity in a cell-specific and promoter-dependent manner. PMID- 9203635 TI - Molecular cloning and pharmacological characterization of a molluscan octopamine receptor. AB - We describe the cloning and functional expression of a cDNA encoding a novel G protein-coupled receptor, which was isolated from the central nervous system of the pond snail Lymnaea stagnalis. The amino acid sequence predicted by this cDNA shows highest similarity with the sequence of the Locusta tyramine receptor, the Drosophila tyramine/octopamine receptor, and the mammalian alpha-adrenergic receptors. On expression in mammalian cells, [3H]rauwolscine, an alpha2 adrenergic receptor antagonist, binds with high affinity (K(D) = 2.9 x 10(-9) M) to the receptor. Of several tested neurotransmitters, octopamine (which is considered to be the invertebrate counterpart of norepinephrine) showed the highest affinity (1.9 x 10(-6) M) for the receptor. Therefore, we consider this receptor to be the first true octopamine receptor to be cloned. The ligand binding properties of the novel receptor, designated Lym oa1, seem to be distinct from any of the binding profiles described for octopamine receptors in tissue preparations. Although the pharmacological profile of Lym oa1 shows some similarity with that of Tyr/Oct-Dro and Tyr-Loc, there are also clear differences. In particular, phentolamine, chlorpromazine, and mianserine display markedly higher affinities for Lym oa1 than for the insect receptors. As far as the vertebrate adrenergic receptors are concerned, the ligand binding properties of Lym oa1 resemble alpha2-adrenergic receptors more than they do alpha- or beta adrenergic receptors. Octopaminergic stimulation of Lym oa1 induces an increase in both inositol phosphates and cAMP (EC50 = 9.1 x 10(-7) M and 5.1 x 10(-6) M, respectively). This is in contrast to the signal transduction pathways described for the related tyramine- and alpha2-adrenergic receptors, which couple in an inhibitory way to adenylyl cyclase. PMID- 9203636 TI - Dual agonistic and antagonistic property of nonpeptide angiotensin AT1 ligands: susceptibility to receptor mutations. AB - Two nonpeptide ligands that differ chemically by only a single methyl group but have agonistic (L-162,782) and antagonistic (L-162,389) properties in vivo were characterized on the cloned angiotensin AT1 receptor. Both compounds bound with high affinity (K(I) = 8 and 28 nM, respectively) to the AT1 receptor expressed transiently in COS-7 cells as determined in radioligand competition assays. L 162,782 acted as a powerful partial agonist, stimulating phosphatidylinositol turnover with a bell-shaped dose-response curve to 64% of the maximal level reached in response to angiotensin II. Surprisingly, L-162,389 also stimulated phosphatidylinositol turnover, albeit only to a small percentage of the angiotensin response. The prototype nonpeptide AT1 agonist L-162,313 gave a response of approximately 50%. The apparent EC50 values for all three compounds in stimulating phosphatidylinositol turnover were similar, approximately 30 nM, corresponding to their binding affinity. Each of the three compounds also acted as angiotensin antagonists, yet in this capacity the compounds differed markedly, with IC50 values ranging from 1.05 x 10(-7) M for L-162,389 to 6.5 x 10(-6) for L 162,782. A series of point mutations in the transmembrane segments (TMs) of the AT1 receptor had only minor effect on the binding affinity of the nonpeptide compounds, with the exception of A104V at the top of TM III, which selectively impaired the binding of L-162,782 and L-162,389. Substitutions in the middle of TM III, VI, or VII, which did not affect the binding affinity of the compounds, impaired or eliminated the agonistic efficacy of the nonpeptides but with only minor or no effect on the angiotensin potency or efficacy. Thus, in the N295D rat AT1 construct, L-162,782, L-162,313, and L-162,389 all antagonized the angiotensin-induced phosphatidylinositol turnover with surprisingly similar IC50 values (90-180 nM), and they all bound with unaltered, high affinity (22-36 nM). However, L-162,313 and L-162,782 could stimulate phosphatidylinositol turnover to only 20% of that of angiotensin. It is concluded that minor chemical modifications of either the compound or the receptor can dramatically alter the agonistic efficacy of biphenyl imidazole compounds on the AT1 receptor without affecting their affinity, as determined in binding assays, and that a number of substitutions in the middle of the TM segments affect the efficacy of nonpeptide agonists as opposed to angiotensin. PMID- 9203637 TI - Pharmacology of muscarinic receptor subtypes constitutively activated by G proteins. AB - We have examined the effects of raising G protein concentration on the pharmacology of a series of agonist and antagonist ligands at the m1, m3, and m5 muscarinic subtypes using a functional assay. Overexpression of G(alpha q) induced constitutive activity of these receptors. The constitutive activity was reversed completely by every muscarinic antagonist tested, which indicates that they are all negative antagonists (inverse agonists). The potencies of antagonists for reversing G protein-induced activity and agonist-induced activity were identical, suggesting the same mechanism of action. Overexpression of G(alpha q) increased the potencies of every tested agonist and the efficacies of all partial agonists. The fold-gains in potency were positively correlated with ligand efficacy with the most efficacious agonists displaying the greatest potency gains. In addition, the efficacies of partial agonists approached those of full agonists. Constitutive activity of receptors has been explained by allosteric models in which receptors exist in spontaneous equilibrium between active and inactive conformations that are stabilized by agonists and antagonists, respectively. In this context, drug efficacy and potency are interrelated because they both depend on the same parameters, namely the absolute and relative affinities of a compound for receptors in active and inactive states and the ratio and concentrations of receptors in active and inactive states. All of our data are consistent with this model, in which raising G protein levels favors formation of the active conformation of receptors. Based on our findings, regulation of G protein concentration may be an important means of controlling receptor activity in vivo. These results define the functional relationship between G protein levels and muscarinic receptor pharmacology. PMID- 9203638 TI - "Orphan" alpha6 nicotinic AChR subunit can form a functional heteromeric acetylcholine receptor. AB - Previously, a rat brain cDNA was reported that was designated alpha6 because of its homology with nicotinic acetylcholine receptor (AChR) alpha subunits, being especially similar to alpha3, but no acetylcholine-gated cation channels were detected when it was expressed in Xenopus laevis oocytes alone or in combination with other known rat AChR subunits. We cloned chicken alpha6 and human beta4 AChR subunits and tested for acetylcholine-gated cation channels with alpha6 by expression in X. laevis oocytes alone or in pairwise combination with chicken alpha3, beta2, or beta4 or with human alpha3, beta2, or beta4 AChR subunits. Chicken alpha6 formed detectable functional AChRs only when expressed together with the human beta4 subunit. The alpha6beta4 AChR-mediated currents show strong inward rectification and dependence on extracellular Ca2+. It exhibited a distinct pharmacological profile with an EC50 value of 28 microM for acetylcholine, 24 nM for (+)-epibatidine, 6.6 microM cytisine, and 15 microM 1,1 dimethyl-4-phenylpiperazinium. Both cytisine and 1,1-dimethyl-4 phenylpiperazinium behaved as partial (approximately 30%) agonists. Remarkably, nicotine (EC50 = 22 microM) was an even weaker partial agonist (approximately 18%) and had a relatively long-lasting inhibitory effect. Coexpression of the previously cloned rat alpha6 subunit with the human the beta4 subunit also resulted in functional alpha6beta4 AChRs with properties resembling those of the chicken/human alpha6beta4 AChRs. Therefore, alpha6 can function as part of AChRs with unusual pharmacological properties. PMID- 9203639 TI - Contrasting actions of lanthanum on different recombinant gamma-aminobutyric acid receptor isoforms expressed in L929 fibroblasts. AB - Functional studies have indicated that, unlike most divalent cations, lanthanum increases both native and recombinant gamma-aminobutyric acid (GABA) receptor (GABAR) currents. In the present study, we have examined whether lanthanum shows subunit-dependent selectivity for modification of currents from different GABAR isoforms. The effects of lanthanum on three different GABAR isoforms, alpha1beta3gamma2L, alpha6beta3gamma2L, and alpha6beta3delta, were determined by transient expression of combinations of alpha1, alpha6, beta3, gamma2L, and delta subunit cDNAs in L929 fibroblasts. Whole-cell recording was used to determine the concentration-response curves for lanthanum for the three different isoforms at submaximal concentrations of GABA. Lanthanum displayed strong potentiation of alpha1beta3gamma2L GABAR currents consistent with earlier reports of potentiation of GABAR currents by lanthanum in neurons and recombinant GABAR isoforms. However, in contrast to the potentiation of alpha1beta3gamma2L GABAR currents by lanthanum, alpha6beta3delta GABAR currents were strongly inhibited and alpha6beta3gamma2L GABAR currents were weakly inhibited by lanthanum. Interaction of lanthanum with GABAR isoforms was competitive, with lanthanum decreasing the EC50 value for GABA of alpha1beta3gamma2L GABARs without changing the maximum current and increasing the EC50 value for GABA of alpha6beta3delta and alpha6beta3gamma2L GABAR currents (greater shift in EC50 value in the alpha6beta3delta compared with the alpha6beta3gamma2L GABARs) without changing the maximum GABAR current. Neither potentiation nor inhibition of GABAR currents by lanthanum showed any voltage dependence. These results suggest that 1) changing the alpha-subunit subtype from alpha1 to alpha6 altered the effect of lanthanum from potentiation to inhibition, 2) changing the gamma2L subunit to the delta-subunit changed the level of maximal inhibition of alpha6 subtype containing GABAR currents by lanthanum, and 3) the site for interaction with lanthanum probably was on the extracellular surface of GABARs. PMID- 9203640 TI - Determinants of specificity for alpha-conotoxin MII on alpha3beta2 neuronal nicotinic receptors. AB - The competitive antagonist alpha-conotoxin-MII (alpha-CTx-MII) is highly selective for the alpha3beta2 neuronal nicotinic receptor. Other receptor subunit combinations (alpha2beta2, alpha4beta2, alpha3beta4) are >200-fold less sensitive to blockade by this toxin. Using chimeric and mutant subunits, we identified amino acid residues of alpha3 and beta2 that participate in determination of alpha-CTx-MII sensitivity. Chimeric alpha subunits, constructed from the alpha3 and alpha4 subunits, as well as from the alpha3 and alpha2 subunits, were expressed in combination with the beta2 subunit in Xenopus laevis oocytes. Chimeric beta subunits, formed from the beta2 and beta4 subunits, were expressed in combination with alpha3. Determinants of alpha-CTx-MII sensitivity on alpha3 were found to be within sequence segments 121-181 and 181-195. The 181-195 segment accounted for approximately half the difference in toxin sensitivity between receptors formed by alpha2 and alpha3. When this sequence of alpha2 was replaced with the corresponding alpha3 sequence, the resulting chimera formed receptors only 26-fold less sensitive to alpha-CTx-MII than alpha3beta2. Site directed mutagenesis within segment 181-195 demonstrated that Lys185 and Ile188 are critical in determination of sensitivity to toxin blockade. Determinants of alpha-CTx-MII sensitivity on beta2 were mapped to sequence segments 1-54, 54-63, and 63-80. Site-directed mutagenesis within segment 54-63 of beta2 demonstrated that Thr59 is important in determining alpha-CTx-MII sensitivity. PMID- 9203641 TI - The long cytoplasmic carboxyl terminus of the prostaglandin E2 receptor EP4 subtype is essential for agonist-induced desensitization. AB - The 488-amino acid human prostaglandin E2 receptor EP4 subtype, which couples to stimulation of adenylyl cyclase, shares the major structural features of G protein-coupled receptors, having seven putative transmembrane domains, an extracellular amino terminus, and a cytoplasmic carboxyl terminus. The latter is composed of 156 amino acids and contains 38 serine and threonine residues, which are potential phosphorylation sites. The carboxyl terminus may be important in receptor function; in some receptors, truncation of the cytoplasmic tail abolishes desensitization. In others, truncation leads to constitutive activity, and in other receptors, truncation has no effect on receptor function. To investigate the role of the long cytoplasmic tail of the EP4 receptor, we constructed a mutant EP4 that lacks the last 138 amino acids at the carboxyl terminus, including 36 serine and threonine residues. The truncated EP4 receptor was stably expressed in Chinese hamster ovary cells at levels comparable to that of the wild-type receptor and exhibited a Kd value for [3H]PGE2 binding similar to that of the wild-type receptor. PGE2-mediated adenylyl cyclase activity as a function of PGE2 concentration was similar in cells expressing the wild-type and truncated EP4 receptors. Neither the wild-type receptor nor the truncated form showed any constitutive activity. However, the wild-type EP4 receptor underwent PGE2-induced desensitization fully within 15-20 min, whereas the truncated EP4 receptor, lacking 36 of the 38 carboxyl-terminal serines and threonines, displayed a sustained activation. Despite the continuous presence of PGE2, the rate of cAMP synthesis via stimulation of the truncated receptor remained constant over > or = 20 min. These findings suggest that the cytoplasmic tail of EP4 plays an important role in agonist-induced desensitization. PMID- 9203642 TI - Binding of a thrombin receptor tethered ligand analogue to human platelet thrombin receptor. AB - A thrombin receptor-radioligand binding assay was developed using [3H]A(pF F)R(ChA)(hR)Y-NH2 ([3H]haTRAP), a high affinity thrombin receptor-activating peptide (TRAP), and human platelet membranes. Scatchard analysis of saturation binding data indicated that [3H]haTRAP bound to platelet membranes with a Kd of 15 nM and a Bmax of 5.2 pmol/mg of protein. The binding was reduced by GPPNHP, a nonmetabolizable GTP analogue. Various TRAPs and a TRAP antagonist, but not other receptor agonists, displaced [3H]haTRAP from the binding sites. SFLLRN-NH2, a thrombin receptor-tethered ligand analogue, and [3H]haTRAP exhibited competitive binding for the same binding sites. The relative affinity of these peptides for the binding site paralleled their EC50 or IC50 values for platelet aggregation. These data indicate that [3H]haTRAP binds specifically and saturably to the functioning G protein-linked thrombin (tethered ligand) receptor in human platelet membranes. PMID- 9203643 TI - Perspective: validating surrogate markers--are we being naive? AB - Because of the difficulties in conducting studies of clinical efficacy of new therapies for human immunodeficiency virus infection and other diseases, there is increasing interest in using measures of biologic activity as surrogates for clinical end points. A widely used criterion for evaluating whether such measures are reliable as surrogates requires that the putative surrogate fully captures the "net effect"-the effect aggregated over all mechanisms of action-of the treatment on the clinical end point. The variety of proposed metrics for evaluating the degree to which this criterion is met are subject to misinterpretation because of the multiplicity of mechanisms by which drugs operate. Without detailed understanding of these mechanisms, metrics of "surrogacy" are not directly interpretable. Even when all of the mechanisms are understood, these metrics are associated with a high degree of uncertainty unless either treatment effects are large in moderate-size studies or sample sizes are large in studies of moderately effective treatments. PMID- 9203644 TI - Longitudinal analysis of quantitative virologic measures in human immunodeficiency virus-infected subjects with > or = 400 CD4 lymphocytes: implications for applying measurements to individual patients. National Institute of Allergy and Infectious Diseases AIDS Vaccine Evaluation Group. AB - The natural variability of quantitative virologic measures among human immunodeficiency virus (HIV) type 1-infected persons was prospectively studied in 29 untreated persons with >600 CD4 cells/microL and in 15 persons receiving zidovudine monotherapy who had 400-550 CD4 cells/microL at study entry. Cell- and plasma-associated infectious HIV-1, provirus, and virion RNA were determined monthly as were numbers of CD4 and CD8 cells. HIV-1 replication varied widely among subjects with similar CD4 cell counts. The within-individual variability was significantly less than the variability between subjects for all virologic measures. Plasma virion HIV-1 RNA levels had the least variability. A mathematical model was devised to assess whether a potential therapeutic intervention significantly alters peripheral HIV-1 load. The model indicated that three measurements of plasma RNA would be outside the 95th percentile for the expected change in an individual due to natural variability. This approach can be used to accurately assess a therapeutic intervention among persons with low plasma HIV-1 titers. PMID- 9203646 TI - Mucosal immune responses in four distinct compartments of women infected with human immunodeficiency virus type 1: a comparison by site and correlation with clinical information. AB - Because mucosal immune responses may be important in protection against human immunodeficiency virus type 1 (HIV-1), HIV-1-specific immune responses at mucosal sites in natural infection were compared. Total antibody concentrations and HIV-1 specific binding antibody responses in four distinct mucosal sites and serum were assessed in 41 HIV-infected and 19 HIV-seronegative women. HIV-1 gp160-specific IgG responses were detected in >99% of mucosal samples in infected subjects, with the highest titers in genital secretions. HIV-1-specific IgA was detected in the majority of endocervical secretions (94%) and nasal washes (95%) but less often in vaginal washes (51%) and parotid saliva (38%). There was no significant correlation between mucosal immune response and most clinical factors. Based on methodologic considerations, frequencies of detection, and HIV-1-specific responses, nasal washes and genital secretions may each provide important measures of HIV-1-specific mucosal immune responses in infected women. PMID- 9203645 TI - A randomized trial (ISS 902) of didanosine versus zidovudine in previously untreated patients with mildly symptomatic human immunodeficiency virus infection. AB - In this multicenter study (ISS 902), 554 previously untreated patients with <500 CD4 cells/mm3 and mildly symptomatic human immunodeficiency virus disease were randomized to receive zidovudine or didanosine (ddI). After a mean follow-up of 20 months, 80 patients (40 zidovudine, 40 ddI) had died and 146 had at least one AIDS-defining event (73 zidovudine, 73 ddI). Overall, no difference was found between treatments with respect to progression to AIDS or death. The analysis of relative risk (RR) of progression over time, however, showed an initially minor risk for zidovudine patients and an inversion in the zidovudine-ddI RR in the second and third years of follow-up. Didanosine showed a greater effect on CD4 cell count response. The two drugs confirmed the toxicity patterns already reported in other trials, with a low occurrence of pancreatitis (ddI 1.3%, zidovudine 0.4%). The overall results suggest that, in this population, zidovudine and ddI monotherapies have comparable long-term clinical efficacy and that more powerful regimens should be preferred. PMID- 9203647 TI - Detection of clonal T cell populations with closely related T cell receptor junctional sequences in persons at high risk for human immunodeficiency virus (HIV) infection and in patients acutely infected with HIV. AB - The T cell repertoires were characterized for CD4+ and CD4 lymphocytes derived from 2 patients with acute human immunodeficiency virus (HIV) infection and from 25 HIV-seronegative persons at high risk for acquiring HIV. Oligoclonal expansions of CD4 cells were detected in the HIV-infected patients and in 2 of 3 uninfected high-risk subjects with a reduced number of CD4+ lymphocytes. Furthermore, nucleotide sequencing revealed that some of the T cell receptor (TCR) beta variable segments (TCRBV), which were highly selected in the high-risk subjects, shared closely related junctional sequences, with the TCRBV predominantly expanded in the HIV-infected patients. Since the likelihood that these similarities occurred by chance is extremely low, these data provide direct molecular evidence in support of several cellular and serologic studies suggesting that some persons remain uninfected despite exposure to HIV. PMID- 9203648 TI - Safety and pharmacokinetics of hyperimmune anti-human immunodeficiency virus (HIV) immunoglobulin administered to HIV-infected pregnant women and their newborns. Pediatric AIDS Clinical Trials Group Protocol 185 Pharmacokinetic Study Group. AB - The pharmacokinetics and safety of hyperimmune anti-human immunodeficiency virus (HIV) intravenous immunoglobulin (HIVIG) were evaluated in the first 28 maternal infant pairs enrolled in a randomized, intravenous immunoglobulin (IVIG) controlled trial of HIVIG maternal-infant HIV transmission prophylaxis. Using 200 mg/kg, mean half-life and volume of distribution (Vd) in women were 15 days and 72 mL/kg, respectively, after one and 32 days and 154 mL/kg after three monthly infusions, with stable 4 mL/kg/day clearance. Transplacental passage occurred. Newborn single-dose half-life, Vd, and clearance were 30 days, 143 mL/kg, and 4 mL/kg/day, respectively. HIVIG rapidly cleared maternal serum immune complex dissociated p24 antigen, and plasma HIV-1 RNA levels were stable. Mild to moderate adverse clinical effects occurred in 2 of 103 maternal and 2 of 25 infant infusions. No adverse hematologic, blood chemistry, or immunologic effects were seen. HIVIG is well-tolerated in HIV-infected pregnant women and their newborns, clears antigenemia, crosses the placenta, and exhibits pharmacokinetics similar to those of other immunoglobulin preparations. PMID- 9203649 TI - Proteosomes, emulsomes, and cholera toxin B improve nasal immunogenicity of human immunodeficiency virus gp160 in mice: induction of serum, intestinal, vaginal, and lung IgA and IgG. AB - Intranasal immunization of mice with human immunodeficiency virus (HIV) rgp160 complexed to proteosomes improved anti-gp160 serum IgA and IgG titers, increased the number of gp160 peptides recognized, and stimulated anti-gp160 intestinal IgA compared with immunization with uncomplexed rgp160 in saline. These enhanced responses were especially evident when either a bioadhesive nanoemulsion (emulsomes) or cholera toxin B subunit (CTB) was added to the proteosome-rgp160 vaccine. Furthermore, anti-gp160 IgG and IgA in vaginal secretions and fecal extracts were induced after intranasal immunization with proteosome-rgp160 delivered either in saline or with emulsomes. Formulation of uncomplexed rgp160 with emulsomes or CTB also enhanced serum and selected mucosal IgA responses. Induction of serum, vaginal, bronchial, intestinal, and fecal IgA and IgG by intranasal proteosome-rgp160 vaccines delivered in saline or with emulsomes or CTB is encouraging for mucosal vaccine development to help control the spread of HIV transmission and AIDS. PMID- 9203650 TI - Utility of urine and leukocyte cultures and plasma DNA polymerase chain reaction for identification of AIDS patients at risk for developing human cytomegalovirus disease. AB - Urine and blood leukocyte cultures and qualitative plasma polymerase chain reaction (PCR) and quantitative competitive (QC) PCR were evaluated for their ability to identify AIDS patients at risk for human cytomegalovirus (HCMV) disease. AIDS patients were followed with urine and blood specimens every 3 months. During a mean follow-up of 12 months, 26 (28%) developed HCMV disease. The sensitivity, specificity, positive predictive value, and negative predictive value for urine culture were 85%, 29%, 31%, and 83%; for leukocyte culture were 38%, 74%, 69%, and 81%; for qualitative plasma PCR were 89%, 75%, 58%, and 94%; for QC-PCR (>1000 copies/microL) were 35%, 100%, 100%, and 80%; and for QC-PCR (>100 copies/microL) were 73%, 90%, 73%, and 90%, respectively. Of 41 patients identified by qualitative PCR to have HCMV DNA in plasma, the 24 who developed HCMV disease had 1510 +/- 448 (mean +/- SE) peak copies of HCMV DNA/microL by QC PCR, versus 161 +/- 52 for the 17 patients who did not develop disease (P = .0007). Thus, plasma PCR is superior to culture for identification of AIDS patients at risk for HCMV disease, and quantitation of plasma DNA further identifies high-risk persons. PMID- 9203651 TI - Murine cytomegalovirus replication in the lungs of athymic BALB/c nude mice. AB - Murine cytomegalovirus (MCMV) infection in the lungs of T cell-deficient athymic BALB/c (Nu/ Nu) mice and their immunocompetent heterozygous (Nu/+) littermates was examined. Following intranasal inoculation, MCMV replicated in the lungs of both Nu/Nu and Nu/+ mice, but virus titers were significantly higher in T cell deficient mice. After subcutaneous inoculation, virus disseminated to lung tissue of athymic mice, leading to progressive MCMV replication in lungs that was not seen in the immunocompetent mice. Athymic mice failed to develop an antibody response to MCMV. Histologically, athymic mice uniformly developed focal interstitial cellular aggregates adjacent to blood vessels or airways, which progressively enlarged and coalesced. Pneumonitis was not seen in the lungs of any Nu/+ mice. Thus, MCMV can replicate in the lungs without pneumonitis in immunocompetent mice, but MCMV produces a progressive focal pneumonitis during deficiency of T cell-mediated immunity. PMID- 9203652 TI - Full or partial seroreversion in patients infected by hepatitis C virus. AB - Cases of partial seroreversion have been reported in hemodialyzed or immunodepressed patients, but spontaneous clearance of viremia associated with a disappearance of specific antibodies or clearance while receiving therapy has not been precisely documented in immunocompetent hepatitis C virus (HCV)-infected persons. A longitudinal study of markers of HCV infection in a cohort of 178 multitransfused patients followed over an 8-year period was done to establish well-documented cases of partial or full seroreversion. Thirty (16.8%) of 178 patients were HCV-infected; among them, 5 had partial or full seroreversion. Seroreversion to an anti-HCV-negative state is characterized by a quantitative decrease in antibody. A seroreversion may be observed in three circumstances: spontaneously, induced by therapy, and in conjunction with human immunodeficiency virus infection. Long-term follow-up of seroreverters will establish whether they have definitively eradicated HCV from their systems. PMID- 9203653 TI - Rhinoviruses induce interleukin-8 mRNA and protein production in human monocytes. AB - Rhinoviruses are important upper respiratory pathogens that are strongly associated with asthma exacerbations. However, the inflammatory response to rhinovirus infection is poorly understood. Interleukin (IL)-8 has been implicated in the pathogenesis of respiratory viral infections and asthma. Rhinovirus induced IL-8 release and mRNA induction were examined in peripheral blood mononuclear cells (PBMC). Rhinoviruses induced IL-8 release for up to 7 days after inoculation onto PBMC. This was associated with an increase in IL-8 mRNA expression that peaked 48 h after exposure to the virus. IL-8 protein production was reduced by UV inactivation of the virus and abolished by preventing virus receptor binding. Although rhinovirus replication was not demonstrated in PBMC, low-grade productive infection was shown in the human monocyte cell line THP-1. Rhinovirus induction of IL-8 in monocytes or airway macrophages may be important in the pathogenesis of rhinovirus-induced asthma exacerbation. PMID- 9203654 TI - Heterotypic protection following oral immunization with live heterologous rotaviruses in a mouse model. AB - A Jennerian approach using live animal viruses to immunize humans is the current lead strategy for developing rotavirus vaccines. This strategy has been modified by incorporating human rotavirus VP7 genes into vaccine strains to induce serotype-specific neutralizing antibodies to human strains. However, the role of homotypic versus heterotypic immunity in protection is unclear. To investigate the importance of serotype-specific immunity in a mouse model, mice were immunized with rhesus rotavirus (RRV: G3, P5[3]), RRV-based modified Jennerian vaccine strains DxRRV (G1, P5[3]), DS1xRRV (G2, P5[3]), or ST3xRRV (G4, P5[3]), or bovine rotavirus NCDV (G6, P6[1]) and challenged with murine rotavirus ECw (G3, P[16]). Mice immunized with modified Jennerian vaccines exhibited complete to near-complete protection from challenge. NCDV-immunized mice also showed partial protection. The protection was correlated with fecal IgA levels to VP6, not serum IgG responses. Modified Jennerian vaccines induce both heterotypic and homotypic immunity in mice. PMID- 9203655 TI - Humoral and cellular immune responses following vaccination with purified recombinant hemagglutinin from influenza A (H3N2) virus. AB - Adults were immunized with either baculovirus-expressed, purified recombinant hemagglutinin (rHA) from influenza A/Beijing/32/92 (H3N2) virus or saline placebo and evaluated for humoral and in vitro cellular immune responses. Compared with responses in placebo recipients, vaccinees had greater postvaccination H3(Beijing/32) HA (H3)-specific lymphoproliferation and interleukin (IL)-2, IL 10, and interferon-gamma (IFN-gamma) production. Mean increases in the production of IL-10 (> or = 20-fold) and IL-2 (10-fold) were relatively greater than that of IFN-gamma (4-fold) or IL-4 (no change). Serum H3 antibodies were induced in 80% of rHA recipients, and the rise in antibody titer was significantly correlated with changes in IL-2, IL-10, and IFN-gamma concentrations. Vaccination with rHA only minimally enhanced anti-influenza virus cytotoxic T lymphocyte activity. These data demonstrate that rHA immunization of adults elicits a significant recall response by memory B and T lymphocytes and suggest that the cytokine response to vaccination has a T helper cell type 0-like profile. PMID- 9203656 TI - Adjuvant activity of the heat-labile enterotoxin from enterotoxigenic Escherichia coli for oral administration of inactivated influenza virus vaccine. AB - Alternative strategies for vaccination against influenza that elicit both systemic antibody and mucosal IgA responses are needed to improve the efficacy in protection against infection. This study demonstrated that oral delivery of inactivated influenza vaccine with the heat-labile enterotoxin (LT) from enterotoxigenic Escherichia coli elicited the spectrum of humoral and cell mediated responses in BALB/c mice critical for the protection and recovery from influenza virus infection. Coadministration of LT with oral influenza vaccine increased antiviral serum IgG and mucosal IgA responses compared with administration of oral influenza vaccine alone. Serum hemagglutination-inhibition and neutralizing antibodies were also augmented by LT. The adjuvant potentiated protection from infection with influenza A H3N2 viruses in mouse lower and upper respiratory tracts, enabling the use of lower doses of oral vaccine. Coadministration of LT with oral inactivated influenza vaccine induced influenza virus-specific proliferative T cells, interleukin-2 production, and major histocompatibility complex class I-restricted cytotoxic T cells. PMID- 9203657 TI - Preclinical evaluation of a novel group B meningococcal conjugate vaccine that elicits bactericidal activity in both mice and nonhuman primates. AB - Group B meningococcal (GBM) conjugate vaccines were prepared using chemically modified N-propionylated polysialic acid, from Escherichia coli K1 polysaccharide capsule, coupled by reductive amination to tetanus toxoid and purified recombinant GBM porin (rPorB). All conjugates elicited high antibody levels in mice with good booster responses. However, only rPorB conjugates elicited bactericidal activity specific against a broad spectrum of five different GBM serotypes. Bactericial activity was completely inhibited by free N-propionylated polysaccharide. In baboons and rhesus monkeys, rPorB conjugates elicited high antibody titers, with IgG booster responses 9- to 15-fold higher than primary responses. Bactericial activity increased 19- to 39-fold over preimmune values, using rabbit complement; increased bactericial activity was also confirmed with human and monkey complement. IgG cross-reactivity for unmodified N-acetyl polysaccharide was <5% for 79% of mice and <10% for 80% of primates. These studies strongly suggest that the N-propionylated polysialic acid-rPorB conjugate is an excellent vaccine candidate for human use. PMID- 9203658 TI - Receptor specificities of variant Gal(alpha1-4)Gal-binding PapG adhesins of uropathogenic Escherichia coli as assessed by hemagglutination phenotypes. AB - Receptor specificities of the three recognized variants of PapG, the major adhesin of uropathogenic Escherichia coli, have been deduced in part from the distinctive hemagglutination (HA) patterns these adhesins ostensibly exhibit with different erythrocytes. A comprehensive reevaluation of these HA patterns using sheep, rabbit, and diverse human erythrocytes revealed that the erythrocyte binding ranges of the three PapG variants are broader and more overlapping than generally recognized, suggestive of receptor specificities beyond those posited in the prevailing model. Experiments involving guinea pig erythrocytes artificially labeled with defined Gal(alpha1-4)Gal-containing glycolipids supported this hypothesis and further suggested that HA patterns with naturally occurring erythrocytes cannot be used to determine reliably an adhesin's glycolipid binding specificities. Finally, slide HA assays exhibited considerable interexperiment, interobserver, and intraobserver irreproducibility, limiting their usefulness for assessing either the receptor specificities of PapG variants or the PapG repertoire of wild-type E. coli strains. PMID- 9203659 TI - Epitope repertoire of human CD4+ T cells on tetanus toxin: identification of immunodominant sequence segments. AB - Sequence regions of tetanus toxin-forming CD4+ cell epitopes in 8 HLA-disparate subjects were identified. Overlapping synthetic peptides corresponding to the complete tetanus toxin sequence were used to test, in a proliferation assay, unselected blood CD4+ cells or CD4+ cell lines propagated by stimulation with tetanus toxoid. The CD4+ cell lines recognized most peptides recognized by the blood CD4+ cells and they recognized additional peptides. Their responses were stronger than those of unselected blood CD4+ cells. Two peptides were recognized by all subjects: one largely overlapped a tetanus toxin sequence region previously identified as a "universal" T cell epitope. Thirteen other peptides elicited a CD4+ cell response in 6 or 7 of the 8 subjects, and another 10 elicited responses in 5 subjects. PMID- 9203660 TI - Molecular comparison of group A streptococci of T1M1 serotype from invasive and noninvasive infections in Finland. AB - A total of 98 isolates of group A streptococci of T1M1 serotype isolated in Finland during 1988-1995 from bacteremic, pharyngeal, and pyogenic infections were studied by molecular means. The typing techniques used included restriction endonuclease analysis, ribotyping, and random amplified polymorphic DNA analysis. Representatives of each observed genotype were also examined for carriage of the speA gene encoding for pyrogenic exotoxin A. All of the serotype T1M1 isolates studied were considered to be of a single clonal origin, and 66% were identical by all three genotyping techniques used. Among the remaining 34% of closely related isolates, 13 different genotypes were detected. All isolates studied carried the speA gene. No evidence was found of correlation of the genomic type to the severity of the infection or the time of isolation. PMID- 9203661 TI - Immunization with outer surface protein (Osp) A, but not OspC, provides cross protection of mice challenged with North American isolates of Borrelia burgdorferi. AB - The identification of antigens with the capacity to induce a broad spectrum of protective immunity is an important consideration in the design of a Lyme disease vaccine. In this study, the range of protection provided by outer surface protein (Osp) A or OspC vaccination was compared. Mice actively immunized with OspA or OspC were challenged with 3 North American isolates of Borrelia burgdorferi. OspA immunized mice were fully protected from infection with each of the isolates, whereas mice immunized with OspC were protected from infection with the homologous isolate but not with 2 heterologous isolates. Sequence analysis revealed that the ospA genes from these 3 isolates were >99% homologous, whereas the ospC genes shared only 81%-85% homology. Western blot analysis suggested antigenic heterogeneity associated with OspC but not OspA. These results indicate that genetic and antigenic heterogeneity may limit the usefulness of OspC as a vaccine constituent. PMID- 9203662 TI - Evidence for a commensal, symbiotic relationship between Gardnerella vaginalis and Prevotella bivia involving ammonia: potential significance for bacterial vaginosis. AB - Six strains of Prevotella bivia and 4 of Gardnerella vaginalis were examined for nutrient substrate utilization as part of ongoing studies on the pathogenesis of bacterial vaginosis. Addition of single amino acids to vaginal defined medium (VDM) was stimulatory to the growth of P. bivia but not to G. vaginalis. However, peptides significantly promoted the growth of both organisms. Growth of P. bivia in VDM and VDM supplemented with either amino acids or peptone was accompanied by net ammonia production, while growth of G. vaginalis under the same conditions resulted in net ammonia utilization. Ammonia-enriched supernatants from the growth of P. bivia in peptone-supplemented VDM were stimulatory to G. vaginalis growth. However, ammonia-reduced supernatants from G. vaginalis growth in peptone supplemented VDM had a neutral effect on P. bivia growth. A commensal relationship between P. bivia to G. vaginalis is proposed, with ammonia flow as a mechanism to support this hypothesis. PMID- 9203663 TI - Phospholipase activity in Cryptococcus neoformans: a new virulence factor? AB - Fifty isolates of Cryptococcus neoformans were examined for extracellular phospholipase production after inoculation onto egg yolk agar; 49 produced a pericolonial precipitate indicative of phospholipase activity. Phospholipase B (PLB), lysophospholipase, and lysophospholipase-transacylase activities were identified by radiometric analysis in supernatants from 4 clinical isolates. The ratio of colony diameter to colony plus precipitate on agar (Pz) correlated with PLB activity. Phospholipase production was similar in 12 environmental and 13 clinical isolates of C. neoformans var. gattii. Environmental strains of C. neoformans var. neoformans (n = 8) produced more phospholipase at 72 h than did 17 clinical isolates (mean Pz, 0.57 vs. 0.72; P < .01); however, Pz values were similar at 96 h. Quantitation of cryptococci in the lungs and brains of BALB/c mice inoculated intravenously with 4 strains expressing high, intermediate, or low phospholipase activity revealed a correlation between phospholipase activity and virulence. Phospholipases secreted by C. neoformans may be implicated in virulence. PMID- 9203664 TI - Phagocytosis of viable Candida albicans by alveolar macrophages: lack of opsonin function of surfactant protein A. AB - Surfactant protein A (SP-A) contributes to host defense by opsonizing microbial organisms for phagocytosis by alveolar macrophages (AM). The role of SP-A as opsonin for phagocytosis of Candida albicans was analyzed. AM in suspension exhibited no phagocytosis of nonopsonized yeast. This was not increased by SP-A, whether provided for preincubation of AM or yeast or present during coincubation. However, the engulfment of serum-opsonized yeast by AM in suspension was inhibited by SP-A. This inhibitory effect was mimicked by complement subcomponent C1q and concanavalin A but not by type IV collagen. SP-A did not interfere with phagocytosis of serum-opsonized yeast by adherent AM, monocytes, neutrophils, or peritoneal macrophages. SP-A lacks function as an opsonin for the phagocytosis of C. albicans by AM but interferes with binding of yeast to AM, inhibiting subsequent ingestion. The role of SP-A as an alveolar space opsonin may thus critically depend on the microbial species involved. PMID- 9203665 TI - Bacterial antigen activation of Vdelta1 and Vdelta2 gammadelta T cells of persons infected with human immunodeficiency virus type 1. AB - Vdelta2 gammadelta T cells are readily activated by microbial antigens. In persons infected with human immunodeficiency virus type 1 (HIV-1), the number of gammadelta T cells remains the same or increases in association with reversal of the Vdelta2/Vdelta1 ratio from > or = 1 to < 1. Vdelta2 T cell responses to microbial antigens were tested in 11 HIV-1-infected (> or = 500 CD4 cells/mm3) and 7 uninfected persons. In persons with HIV-1 infection, Mycobacterium tuberculosis expanded Vdelta2 cells in 1 person as did Salmonella typhimurium in 4; however, Candida albicans antigens did not lead to more Vdelta2 cells. Vdelta2 responses to M. tuberculosis were enhanced by interleukin (IL)-2 in HIV-1 infected persons (from 1 subject to 7; P < .01) and were associated with increased interferon-gamma production. Bacterial antigens and IL-2 increased HIV 1 replication; M. tuberculosis antigens induced the greatest increase. Thus, in HIV-1-infected persons with > or = 500 CD4 cells/mm3, Vdelta2 T cell responses to bacterial antigens remain intact. PMID- 9203667 TI - Human T cell lymphotropic virus type I does not increase human immunodeficiency virus viral load in vivo. AB - Human T lymphotropic virus type I (HTLV-I) can increase human immunodeficiency virus (HIV) replication in vitro, and several studies suggest that HTLV-I accelerates the progression of HIV infection. To determine whether HTLV-I enhances HIV replication in vivo, a case-control study was done of serum HIV viral load, using polymerase chain reaction, in 23 subjects with HTLV-I/HIV coinfection and 92 control subjects with HIV single infection. The geometric mean serum RNA level was 11,482 copies/mL in the coinfected group and 13,804 in the single-infection group (P = .57), a result that did not change after adjustment for zidovudine use and CD4 cell count. Among subjects with advanced HIV infection, there was a trend toward higher viral load among singly infected subjects. HTLV-I did not appear to increase HIV plasma RNA levels in subjects with coinfection. These results do not provide a biologic basis for the hypothesis that HTLV-I accelerates the course of HIV infection. PMID- 9203666 TI - The effect on human immunodeficiency virus type 1 RNA levels in cerebrospinal fluid after initiation of zidovudine or didanosine. AB - Human immunodeficiency virus type 1 (HIV-1) RNA, neopterin, and beta2 microglobulin levels were analyzed in cerebrospinal fluid (CSF) and serum before and 3-13 months after initiation of antiretroviral monotherapy in 16 HIV-1 infected persons. Twenty-one treatment periods, 13 after initiation of zidovudine and 8 after initiation of didanosine, were studied. During zidovudine treatment, CSF HIV RNA levels decreased by a mean of 1.05 log10 (-91%, P < .01), and CSF neopterin and beta2-microglobulin levels by 57% and 33%, respectively (P < .01). No reduction was seen during didanosine treatment in CSF HIV RNA (+0.13 log10, not significant), CSF neopterin, or beta2-microglobulin levels. Changes in CSF HIV RNA levels correlated with changes in CSF neopterin and beta2-microglobulin (r(s) = .81 and .83, respectively, P < .001). The decrease in HIV RNA was significantly larger in CSF than in serum following zidovudine treatment (P < .01). These data demonstrate that zidovudine is a potent reducer of central nervous system virus load, which may be important for long-term neuroprotection. PMID- 9203668 TI - Inactivated poliovirus vaccine protects transgenic poliovirus receptor mice against type 3 poliovirus challenge. AB - Transgenic (Tg) mice expressing the human poliovirus receptor (PVR) were vaccinated with inactivated poliovirus vaccine (IPV) and evaluated for induced immunity against type 3 poliomyelitis. One injection of monovalent type 3 IPV elicited protective immunity against wild-type poliovirus. In contrast, 2 injections of trivalent IPV were required for protection. Neutralizing antibody response and protection were vaccine dose-dependent. Administration of polio immune mouse plasma protected unimmunized mice, demonstrating that neutralizing antibody was sufficient for immunity. IPV heated to remove its D antigen component did not induce protection in Tg PVR mice. IPV derived from a wild-type poliovirus strain gave better protection against wild-type viral challenge than IPV derived from an attenuated poliovirus strain. The newly developed Tg PVR mouse-protection test may be useful in evaluating existing IPV potency tests and for attempts to improve formulations of trivalent IPV or combined vaccines for childhood immunization schedules. PMID- 9203669 TI - Dose titration study of live attenuated varicella vaccine in healthy children. Pennridge Pediatric Associates. AB - To approximate the effect of prolonged storage on safety and immunogenicity, healthy children were given a single dose of the currently marketed live attenuated varicella vaccine (3625 pfu) or of a partially heat-inactivated vaccine (1125 or 439 pfu). The 3 doses had similar antigen content (attenuated plus inactive virus particles). The vaccine was well tolerated. No significant differences in adverse reactions were observed. Although the seroconversion rates were excellent at each dose (> or = 98%), the higher doses resulted in significantly greater geometric mean antibody titers at 6 weeks (10.5 and 10.6 ELISA U/mL) compared with the 439 pfu dose (5.7 ELISA U/mL), P < or = .01. One year after immunization, differences in antibodies were similar to the 6-week postimmunization results. Results indicate that until the date of expiry, the vaccine's immunogenicity will be preserved and there will be no clinically important changes in type or frequency of adverse events. PMID- 9203670 TI - Distribution of infecting hepatitis C virus genotypes in end-stage liver disease patients at a large American transplantation center. AB - The distribution of hepatitis C virus (HCV) genotypes was studied in 202 anti-HCV positive liver transplant candidates with end-stage liver disease. HCV sequences were successfully amplified from 185 patients: In the first 100, the genotype was determined by direct sequencing in the NS5 region, and in the remaining 85, type specific primers were used for genotyping. Eighty-five patients (46.0%) were infected with type 1a HCV strains, 52 (28.1%) with type 1b, 14 (7.6%) with type 2b, 13 (7.0%) with type 4, 5 (2.7%) with type 3a, 2 (1.1%) with type 2a, and 1 (0.5%) with type 2c. Thirteen HCV-positive patients (7.0%) could not be genotyped. The relatively low prevalence of genotype 1b in this population of end stage liver disease patients speaks against postulated higher pathogenicity of this genotype. PMID- 9203671 TI - High prevalence of GB virus C/hepatitis G virus in healthy persons in Ho Chi Minh City, Vietnam. AB - GB virus C or hepatitis G virus (GBV-C/HGV), a novel Flavivirus, is detected in 1.5% of US blood donors. The prevalence is higher in multiply transfused patients and in persons with liver disease. Because of the increased incidence of hepatitis in Asia, sera from healthy Vietnamese were tested for the presence of GBV-C/HGV RNA by the reverse transcription polymerase chain reaction. Viral RNA was detected in 5.7% of those tested; 6 of 81 volunteer blood donors had positive samples as did 5 of 97 army recruits and 2 of 50 postpartum women. When the 188 bp product from 6 subjects was sequenced, there was 75%-85% homology at the nucleotide level compared with published sequences, indicating a high degree of genotypic variation, even within a putatively well-conserved region of the viral genome. Viremia with this non-cell-associated novel virus appears to be common among normal persons in Vietnam. PMID- 9203672 TI - Molecular investigation of GB virus C infection in hemophiliacs in Japan. AB - RNA of a putative non-A, -B, -C, -D, or -E hepatitis virus named GB virus C (GBV C) was detected by reverse transcription-polymerase chain reaction with primers deduced from the 5' untranslated region in 15 (24%) of 63 men with hemophilia in Japan at a frequency higher (P < .001) than that in 2 (0.6%) of 337 controls. By phylogenetic analysis, GBV-C isolates from some patients were similar in sequence, indicating infection with closely related strains, and those from certain patients resembled sequences reported from foreign countries. All patients were infected with hepatitis C virus, and genotypes that are rare in Japan were detected in 36 (57%) of them. These results indicate that patients with hemophilia in Japan would be at increased risk for infection with GBV-C and hepatitis C virus, some of which would have been transmitted via imported coagulation factor concentrates in the past. PMID- 9203673 TI - An ELISA for detection of antibodies to the E2 protein of GB virus C. AB - An ELISA was developed for detection of antibodies to GB virus C (GBV-C) using a recombinant E2 protein expressed in CHO cells. Seroconversion to anti-E2 positivity was noted among several persons infected with GBV-C RNA-positive blood through transfusion. Of 6 blood recipients infected by GBV-C RNA-positive donors, 4 (67%) became anti-E2 positive and cleared their viremia. Thus, anti-E2 seroconversion is associated with viral clearance. The prevalence of antibodies to E2 was relatively low (3.0%-8.1%) in volunteer blood donors but was higher in several other groups, including plasmapheresis donors (34.0%), intravenous drug users (85.2%), and West African subjects (13.3%), all of whom tested negative by GBV-C reverse-transcription polymerase chain reaction (RT-PCR). These data demonstrate that testing for anti-E2 should greatly extend the ability of RT-PCR to define the epidemiology and clinical significance of GBV-C. PMID- 9203674 TI - Treatment of experimental methicillin-resistant Staphylococcus epidermidis endophthalmitis with intravitreal vancomycin and intravitreal dexamethasone. AB - The use of intravitreal steroids to treat bacterial endophthalmitis remains controversial. The efficacy of intravitreal vancomycin alone (group 1), intravitreal dexamethasone alone (group 2), and a combination of intravitreal vancomycin and dexamethasone (group 3) in the treatment of experimental methicillin-resistant Staphylococcus epidermidis endophthalmitis was evaluated in a rabbit model: 24 h after bacterial inoculation of all eyes, right eyes were treated and left eyes served as infected controls. Vitreal aspirations and grading of vitreal inflammatory reaction were done regularly until sacrifice. Group 2 eyes demonstrated more inflammation histologically than control eyes. Vitreal aspirations demonstrated no growth by 5 days from groups 1 and 3 eyes. Although the clinical appearance was not significantly different between groups 1 and 3, the histologic appearance of group 3 eyes showed less intense intraocular inflammation. Treatment with dexamethasone in the absence of appropriate antibiotics was more harmful than no treatment at all (P < .05). Therapy with both intravitreal vancomycin and dexamethasone results in less inflammation than intravitreal vancomycin alone in this model. PMID- 9203675 TI - Cross-reactivity to Borrelia burgdorferi proteins in serum samples from residents of a tropical country nonendemic for Lyme disease. AB - Reports of Lyme disease from areas where the disease is not endemic have increased. Eighty-six human serum samples from Papua New Guinea (nonendemic for Lyme disease) were examined for the presence of IgG antibodies that recognize Borrelia burgdorferi antigens, using the currently recommended two-tiered system of analysis (sensitive ELISA with Western blot). The percentage of positive tests dropped from 50% to 10% when individual negative controls were included in the two-tiered analysis. Positive serum samples failed to inhibit the growth of B. burgdorferi in culture and did not yield positive reactions in the fluorescent treponemal antibody-absorption test. These characteristics, together with atypical Western blot antigen recognition patterns and the absence of known vectors, provide evidence that seropositive results for these persons are not the result of exposure to B. burgdorferi. Individual negative controls may minimize false-positive results for serologic tests for Lyme disease, and these tests must be interpreted in the context of clinical and epidemiologic data. PMID- 9203676 TI - Clinical isolates of Shigella species induce apoptosis in macrophages. AB - Shigella species are invasive enterobacteria that cause dysentery, a severe form of diarrhea. The ability to invade epithelial cells and to kill macrophages is essential for virulence in a prototype Shigella flexneri strain. It is shown here that clinical isolates of both S. flexneri and Shigella sonnei invade epithelial cells and are cytotoxic to macrophages in vitro. Furthermore, clinical Shigella strains kill macrophages by inducing apoptosis. The conservation of the ability to induce macrophage apoptosis by clinical isolates suggests that this function plays a crucial role in the pathogenesis of Shigella species. PMID- 9203677 TI - Interleukin-8 and chemotactic activity of middle ear effusions. AB - The importance of interleukin (IL)-8 in the chemotactic activity of middle ear effusions (MEEs) was evaluated. There was a significantly higher IL-8 concentration in MEEs of children with acute otitis media (AOM) (n = 17; 136 ng/mL) than in children with otitis media with effusion (OME) (n = 28; 65 ng/mL). The IL-8 concentration in MEEs with bacteria (149 ng/mL) was significantly higher than in MEEs without bacteria (66 ng/mL). MEEs from children with AOM and OME had equally higher chemotactic activity than the diluent alone (23.3% and 24.8% vs. 9.2%). The chemotactic activity was not altered by the presence of bacteria nor did it correlate with IL-8 concentration. Fractionation of MEEs by gel chromatography demonstrated that the main chemotactic activity could clearly be separated from the IL-8 activity, thus excluding IL-8 as a main chemotactic component in MEEs. PMID- 9203678 TI - Differential production of and migratory response to beta chemokines by human microglia and astrocytes. AB - Little is known about the participation of beta chemokines in inflammatory processes within the central nervous system. The release of three of these peptides (macrophage inflammatory protein [MIP]-1alpha, MIP-1beta, and monocyte chemoattractant protein-1) from human fetal microglial cell and astrocyte cultures was assessed following stimulation by lipopolysaccharide, interleukin 1beta, and tumor necrosis factor-alpha. Although striking differences were found between these two types of glial cells in their responsiveness to lipopolysaccharide and cytokines, both microglia and astrocytes produced all three beta chemokines. Only microglial cells, however, demonstrated an increased migratory response to the beta chemokines. The results of this in vitro study suggest that beta chemokines may play an important role in the trafficking of mononuclear phagocytes within the brain. PMID- 9203679 TI - Dihydropteroate synthase polymorphisms in Pneumocystis carinii. AB - Sulfa drugs are widely used in the treatment and prophylaxis of Pneumocystis carinii pneumonia. The nucleotide sequences of the sulfa target enzyme, dihydropteroate synthase (DHPS), differed substantially in human-, rat-, and mouse-derived P. carinii. Sequence variation also existed in the DHPSs from human derived isolates. Six nucleotide changes were found in 6 human isolates; each was nonsynonymous and resulted in an amino acid change. Several of these changes were in highly conserved regions and are similar to those that cause sulfa resistance in other organisms. These data suggest that the human-derived P. carinii DHPS may be evolving under positive selective pressure from sulfa drugs. PMID- 9203680 TI - Serologic evolution of neurocysticercosis patients after antiparasitic therapy. Cysticercosis Working Group in Peru. AB - Neurocysticercosis is the main cause of acquired epilepsy in developing countries and is an emerging disease in the United States. Introduction of the immunoblot assay provided a new tool for the diagnosis and monitoring of neurocysticercosis. This study analyzed the relationship between clinical characteristics of cerebral infection (number and type of lesions) plus the baseline response on immunoblot and the changes observed after therapy. Reaction to all 7 diagnostic bands was associated with severe infection (more lesions). Seventeen patients (35%) had no active lesions on computed tomography (CT) 3 months after therapy and were considered cured. Although most cured patients remained seropositive after 1 year, 3 became seronegative before 9 months. In these 3 cases, the lesions had resolved on CT at 3 months. Persistent seropositivity does not necessarily indicate active infection. Serologic follow-up will be clinically helpful only in rare cases in which early antibody disappearance occurs. PMID- 9203681 TI - Reduction of Wuchereria bancrofti adult worm circulating antigen after annual treatments of diethylcarbamazine combined with ivermectin in French Polynesia. AB - Circulating filarial antigen (CFA), determined with Og4C3 ELISA, is a marker of Wuchereria bancrofti adult worm infection. The reduction of CFA over 2 years was determined in 185 microfilaremic and 111 amicrofilaremic but CFA+ adults given an annual dose of either diethylcarbamazine (DEC) or ivermectin or the two combined. Reduction of CFA level was good with DEC but weak with ivermectin and followed the same pattern in amicrofilaremic and microfilaremic groups. Combinations and DEC alone had a similar impact on CFA level. CFA clearance was observed in amicrofilaremic but not in microfilaremic persons in all DEC-containing treatments. However, the highest clearance rate was observed in persons treated with DEC at 6 mg/kg combined with ivermectin. Continuous reduction of CFA level after repeated treatments shows that elimination of W. bancrofti infection, monitored by CFA clearance, might be achieved within a few years with annual treatments of DEC combined with ivermectin. PMID- 9203683 TI - Commentary: the unfolding story of global poliomyelitis eradication. PMID- 9203682 TI - IgG4 and IgE responses to Schistosoma mansoni adult worms after treatment. PMID- 9203684 TI - Progress toward global polio eradication. AB - Significant progress is being made towards the global eradication of poliomyelitis by the year 2000. The strategies recommended by the World Health Organization for polio eradication are as follows: maintaining high routine immunization coverage; conducting nationwide mass immunization campaigns; building effective, laboratory-based surveillance for acute flaccid paralysis; and conducting localized immunization campaigns directed at the final reservoirs of virus transmission. Sixty-three countries have conducted nationwide anti-polio immunization campaigns. Three hundred million children were immunized in these campaigns worldwide in 1995. The reported incidence of poliomyelitis has fallen by approximately 80% since the global target was set in 1988, and the geographic range of polio is being restricted. The major challenges for achieving eradication are establishing effective surveillance systems in all countries and mobilizing the resources needed to fully implement the recommended strategies in the 67 countries in which polio remains endemic. PMID- 9203685 TI - Overview of poliomyelitis in the African Region and current regional plan of action. AB - The African Region of the World Health Organization includes a diverse membership of 48 countries and territories that has made substantial progress toward controlling poliomyelitis. The coverage with three doses of oral poliovirus vaccine among 1-year-old children reached 58% in 1995, a substantial increase from 49% in 1993, and the incidence of poliomyelitis decreased from 5126 cases in 1980 to 1597 in 1995. To interrupt poliovirus circulation, 29 countries planned to conduct either national immunization days (25 countries) or subnational immunization days (4 countries) during 1996. To ensure the success of these efforts, high-level political commitment has been obtained in many countries, and the campaign to "Kick polio out of Africa" is supported by some of the most respected African politicians. Provided the necessary resources can be obtained from internal and external sources, the African Region may be able to achieve the eradication of poliomyelitis by the year 2000 or shortly thereafter. PMID- 9203686 TI - Status of polio eradication in the seven countries of the Eastern Africa Epidemiological Block. AB - The progress that Burundi, Eritrea, Kenya, Rwanda, Tanzania, Uganda, and Zambia, the seven countries of the Eastern Africa Epidemiological Block (EAEB), have made toward polio eradication is summarized. Despite low per capita gross national product, poor infrastructure (especially for communication and transportation), and civil unrest, the EAEB has made significant progress toward polio eradication. Five of the seven countries have achieved high levels of routine coverage with at least three doses of oral polio vaccine. Virologic surveillance is established in Tanzania, Uganda, Zambia, and Kenya; will be established in Eritrea in 1997; and will be resumed in Rwanda as soon as civil unrest abates. Political support for polio eradication is strong in the region, and all of the EAEB countries, except Burundi, held either national or subnational immunization days in 1996. PMID- 9203687 TI - Current status of polio eradication in Southern Africa. AB - During the 1990s, poliomyelitis transmission in 11 mainland and island nations of southern Africa appeared relatively low. However, the implementation of specific strategies recommended by the World Health Organization for eradicating polio in southern Africa began only in 1994. In 1995, oral poliovirus vaccine coverage (three doses) among infants was > or = 75% in all but 4 countries. National immunization days (NIDs) to control polio outbreaks were carried out in Namibia in 1994 and 1995. Angola, Botswana, and South Africa carried out subnational NIDs in 1995. All countries in southern Africa except Madagascar planned NIDs in 1996. Epidemiologic surveillance of acute flaccid paralysis (AFP) and laboratory surveillance of wild poliovirus was launched after nationwide training workshops in 7 mainland countries and is planned in the remaining countries by the end of 1996. Analysis of recommended performance indicators of AFP surveillance shows that substantial progress was made during 1994-1995, and the prospects for the certification of polio-free status in southern Africa on target appear good. PMID- 9203688 TI - Poliomyelitis in Angola: current status and implications for poliovirus eradication in Southern Africa. AB - As part of emergency assistance to the Ministry of Health (MOH), national surveillance data for poliomyelitis and charts of cases at the national rehabilitation hospital were reviewed. Poliomyelitis patients admitted to Angola's main pediatric hospital were examined. A mean of 86 cases of poliomyelitis/year were reported in Angola during 1989-1994. Review of records from non-MOH sources uncovered another 74 cases, primarily from areas outside governmental control. Hospital chart reviews revealed that 80% of the cases were children <3 years of age, mainly unvaccinated. Molecular analyses of isolates from cases in Luanda and at the Angola-Namibia border suggest that these isolates are closely related and that > or = 2 strains of wild poliovirus type 1 are circulating currently in Angola. This investigation confirms that poliomyelitis has remained endemic in Angola since independence in 1975. It affects primarily young and unvaccinated children. Control of poliomyelitis in Angola is essential to expand the polio-free zone in southern Africa. PMID- 9203689 TI - Polio outbreaks in Namibia, 1993-1995: lessons learned. AB - In 1993, a nationwide outbreak of 53 cases of paralytic poliomyelitis occurred in Namibia. The World Health Organization-recommended supplemental vaccination strategy of national immunization days (NIDs), providing two doses of oral polio vaccine (OPV) to all children <5 years, was implemented to control the epidemic. A second focal outbreak of 16 confirmed polio cases occurred during 1994-1995 in northeast Namibia. "Mopping-up" vaccination was implemented to control the second outbreak, followed by NIDs. Both epidemics appeared to be associated with wild poliovirus importation from Angola, where polio is endemic. Although supplemental vaccination measures achieved suboptimal OPV coverage, surveillance of acute flaccid paralysis has not detected wild poliovirus in Namibia since April 1995. Future NIDs should aim to ensure OPV coverage >90% in each round of NIDs in each district. Nevertheless, the risk of new poliovirus importations will continue until efforts in Angola to increase routine coverage with three doses of OPV and extend supplemental vaccination activities can be implemented. PMID- 9203690 TI - Eradication of wild poliovirus from the Americas: acute flaccid paralysis surveillance, 1988-1995. AB - In May 1985, the Pan American Health Organization proposed the goal of interruption of wild poliovirus transmission in the Western Hemisphere. An important component of the polio eradication strategy was conducting surveillance for cases of acute flaccid paralysis. Reported cases were thoroughly investigated, including the collection of stool samples for testing for the presence of wild poliovirus. The last patient with poliomyelitis due to wild poliovirus in the Americas had onset of paralysis on 23 August 1991 in Peru. Since then, >9000 cases of acute flaccid paralysis have been reported and thoroughly investigated; none has been confirmed as paralytic poliomyelitis due to wild poliovirus. On 29 September 1994, the International Commission for the Certification of Poliomyelitis Eradication declared the Americas to be polio free. PMID- 9203691 TI - Eradication of wild poliovirus from the Americas: wild poliovirus surveillance- laboratory issues. AB - The Pan American Regional Poliomyelitis Laboratory Network, developed to support the program to eradicate indigenous wild poliovirus transmission in the Americas, included 10 laboratories, distributed in eight countries in the Americas, organized according to the diagnostic procedures they regularly performed. All laboratories isolated and typed virus in stool specimens, several did intratypic differentiation by nucleic acid probe hybridization, and 2 sequenced wild poliovirus isolates for molecular epidemiologic studies. High performance of the network was maintained through comprehensive training of virologists, continuous monitoring of laboratory performance, and prompt investigation of problems. Recommended field and laboratory procedures were regularly reviewed and revised to optimize sensitivity, specificity, and diagnostic efficiency. Close integration of field and laboratory surveillance was achieved through frequent meetings between virologists and epidemiologists, effective communication of program priorities, and the distribution of weekly surveillance reports. PMID- 9203692 TI - Progress toward poliomyelitis eradication in the Eastern Mediterranean Region of the World Health Organization. AB - The Eastern Mediterranean Region (EMR) of the World Health Organization has made substantial progress toward eradicating poliomyelitis. From 1988 to 1995, the number of confirmed cases of polio decreased 66%, from 2342 to 789. National immunization days were conducted in 18 (78%) of the 23 countries in 1995, representing 88% of the regional population. By 1995, 20 countries (87%) in the EMR had established systems for reporting acute flaccid paralysis (AFP), 20 (87%) were investigating AFP or polio cases epidemiologically, 18 (78%) had initiated follow-up at 60 days to confirm or discard suspected polio cases, 7 (30%) had achieved nonpolio AFP rates of > or = 1.0/100,000 children <15 years of age (a measure of the sensitivity of surveillance), and 16 (70%) had made laboratory investigations of polio cases for 1281 (74%) of the 1715 AFP cases reported in the EMR. Despite significant progress, the success of the polio eradication initiative in the EMR will depend on finding solutions to a number of technical, managerial, political, and financial challenges. PMID- 9203693 TI - The eradication of poliomyelitis in Egypt: critical factors affecting progress to date. AB - Poliomyelitis eradication activities in Egypt were reviewed to identify the critical factors for the progress seen by 1995 and to highlight problems that could be avoided in other countries in which poliomyelitis is endemic. National immunization and surveillance data demonstrate that the combination of high routine immunization coverage (>85%) with oral polio vaccine combined with two properly conducted rounds of national immunization days (NIDs) resulted in a 75% reduction in reported polio cases between 1992 and 1993. Available data suggest that earlier control strategies, such as single-round NIDs in 1990 and 1991, the administration of inactivated poliovirus vaccine (IPV) at 2 months of age in 1992 1993, and the use of "mop-up" campaigns while wild poliovirus was still widespread, did not contribute substantially to the recent decline in cases. Proper implementation of the World Health Organization's recommended strategies can eliminate wild poliovirus circulation in the large, densely populated tropical countries in which poliomyelitis remains endemic. PMID- 9203694 TI - Outbreak of paralytic poliomyelitis in a highly immunized population in Jordan. AB - Between November 1991 and March 1992, 37 cases of paralytic poliomyelitis occurred in Jordan, where none had been reported since 1988. Of these, 17 (50%) of 34 patients had received at least three doses of oral poliovirus vaccine (OPV3). The first and 2 subsequent case-patients were children of Pakistani migrant workers, and the first 8 and a total of 27 (75%) case-patients resided in or near the Jordan Valley. A seroepidemiologic study of 987 children in all regions of Jordan was performed to assess OPV3 coverage and immune response to OPV. Although OPV3 coverage by 12 months of age was high (96%) in the general population, coverage was lower among Pakistani (21%), Bedouin (63%), and Gypsy (9%) children (P < .001). Seroprevalences for poliovirus type 3 were 71% in the Jordan Valley versus 81% in other regions after 3 doses of OPV (P < .06) and 77% in the Jordan Valley versus 98% in other regions after 5 doses of OPV (P < .001). This outbreak demonstrates the importance of achieving high seroimmunity to infection in all geographic areas to prevent the reintroduction and spread of imported strains of wild poliovirus. PMID- 9203695 TI - Outbreak of poliomyelitis in Gizan, Saudi Arabia: cocirculation of wild type 1 polioviruses from three separate origins. AB - In 1989, a localized outbreak of 10 cases of poliomyelitis occurred in Saudi Arabia. Wild poliovirus type 1 was isolated from 5 patients. To determine the patterns of poliovirus circulation, partial nucleotide sequences of the poliovirus isolates were compared. These isolates were remarkably diverse. Two isolates were closely related to each other and to viruses isolated during the 1988 epidemic in Oman. Two other isolates were very similar to viruses found in Egypt. The fifth isolate was distantly related to the latter pair. The molecular data suggest that the 10 cases represented three separate outbreaks. The virologic findings underscore the potential for Saudi Arabia, which receives millions of guest workers and their families each year from countries in which polio is endemic, to be exposed to frequent importations of wild polioviruses. To restrict the circulation of imported polioviruses, Saudi Arabia must maintain high population immunity to poliovirus in all geopolitical divisions. PMID- 9203696 TI - Status of poliomyelitis eradication in Europe and the Central Asian republics of the former Soviet Union. AB - In the European Region of the World Health Organization, all countries in which polio is endemic have adopted the following strategies: achievement of high routine vaccination coverage, implementation of supplemental immunization activities, and enhancement of surveillance for poliomyelitis. In 1995, 205 cases of poliomyelitis were reported. Routine coverage among 1-year-olds with three doses of poliovirus vaccine was 89% in 1995. Ten countries conducted national immunization days (NIDs). Twenty-four countries (48%) adopted acute flaccid paralysis (AFP) surveillance. Use of NIDs has decreased poliomyelitis incidence in the seven countries in which polio is endemic (Armenia, Azerbaijan, Kazakhstan, Turkey, Turkmenistan, Tajikistan, Uzbekistan) from 203 cases in 1994 to 47 in 1995, a 77% reduction. Full implementation of the strategies to achieve eradication in the countries in which polio is endemic, including those countries with epidemic poliovirus transmission during 1995, is likely to accomplish regional eradication of poliomyelitis by the year 2000 or earlier. PMID- 9203697 TI - A large outbreak of poliomyelitis following temporary cessation of vaccination in Samarkand, Uzbekistan, 1993-1994. AB - Oral poliovirus vaccine (OPV) was not available in Samarkand, Uzbekistan, from November 1992 to August 1993. The ensuing outbreak of poliomyelitis was investigated: Patients with poliomyelitis were evaluated, the extent of poliovirus circulation was estimated, and the effectiveness of control measures was assessed. Between March 1993 and April 1994, 74 cases of paralytic disease attributable to poliovirus type 3 were reported. Cases originated from 63% of districts; 88% of cases were children < or = 2 years old, and the highest attack rates were for infants 9-11 months (65/100,000) and 12-14 months (60/100,000). Most patients were either unvaccinated (45%) or inadequately vaccinated (23%). Limited quantities of OPV became available in September 1993 and were provided to infants (3 doses) and 1-year-olds (2 doses), controlling rapidly the outbreak in these age groups, but cases continued, primarily among older children, until April 1994. These findings suggest that control efforts should be guided by the age distribution of the children with poliomyelitis. PMID- 9203698 TI - Unimmunized Gypsy populations and implications for the eradication of poliomyelitis in Europe. AB - The certification of poliomyelitis eradication in Europe will eventually require that countries demonstrate there is a minimal risk of wild poliovirus reintroduction and sustained transmission through unimmunized subpopulations such as ethnic minorities. A serologic survey among a Gypsy community in Italy found that despite only 26% documented immunization coverage, serum neutralizing antibodies to poliovirus types 1, 2, and 3 were detected in 81%, 94%, and 63% of the 86 persons studied. While the high level of immunity found in this community may have been due to secondary spread of vaccine virus, the possibility of unrecognized circulation of wild polioviruses cannot be excluded. Targeted immunization of such groups may be the most efficient means of eliminating the risk of importation-associated outbreaks. PMID- 9203699 TI - Polio eradication in the World Health Organization South-East Asia Region by the year 2000: midway assessment of progress and future challenges. AB - In the South-East Asia Region (SEAR) of WHO, paralytic poliomyelitis has decreased from 25,711 cases in 1988 to 3304 cases in 1995, representing an 87% reduction. By 1995, in 6 of 10 member countries--India, Bangladesh, Myanmar, Nepal, Indonesia, and Democratic People's Republic of Korea--polio remained endemic. Two countries, Sri Lanka and Thailand, appear close to polio eradication, and 2, Bhutan and Maldives, reported no cases during 1989-1995. Although reported rates of acute flaccid paralysis and the percentage of cases virologically investigated are low in some countries, no isolates of wild poliovirus type 2 have been reported outside India since 1993. By the end of 1996, all 8 countries in which polio is endemic will have conducted national immunization days for polio eradication. The major challenge for polio eradication in SEAR will be strengthening surveillance, because national immunization days alone cannot eradicate polio. PMID- 9203700 TI - Poliomyelitis eradication in the Western Pacific Region. AB - Polio eradication activities in the Western Pacific Region (WPR) have reduced the transmission of wild poliovirus to one remaining focus of endemic transmission in the Mekong Delta area of South Vietnam and Cambodia. There has been a high level of government commitment for national immunization days in all WPR countries in which poliomyelitis was previously endemic and for continuous improvement in acute flaccid paralysis (AFP) surveillance quality. The total number of reported confirmed poliomyelitis cases in 1995 (as of June 1996) was 432, only 7% of the total of 5825 cases reported in 1990. In 1995, wild poliovirus was isolated from only 19 of 4800 AFP patients from whom specimens were collected and analyzed. There has been one importation of wild poliovirus type 1 into China from a neighboring country. An international Regional Commission for the Certification of Poliomyelitis Eradication in the WPR has been formed and met for the first time in April 1996. PMID- 9203701 TI - Status of the eradication of indigenous wild poliomyelitis in the People's Republic of China. AB - A large nationwide outbreak occurred in 1989-1990 in China, in which nearly 10,000 poliomyelitis cases were reported. After two rounds of oral poliovirus vaccine (OPV) supplemental activity in nearly every province in the 1992-1993 winter season, no wild poliovirus was detected in 1993 in 22 provinces in the middle of China that contained 86% of the population. During the first national immunization days (NIDs) conducted in December 1993 and January 1994, 83 million children 0-47 months of age were immunized. In 1994, wild poliovirus was identified in only 6 of 2397 children with stool specimens tested. After a second NID in December 1994 and January 1995, no wild poliovirus was detected in 1995 despite a very high level of virus surveillance. In summary, double-round mass supplemental OPV immunizations in children 0-3 years old in two consecutive winters eliminated wild poliovirus from 23% of the world's population (1.2 billion people). PMID- 9203702 TI - Poliovirus surveillance: building the global Polio Laboratory Network. AB - A network of virologic laboratories has been established by the World Health Organization to conduct surveillance for wild poliovirus and to provide evidence for the certification of poliomyelitis eradication. The network consists of >60 national laboratories isolating and identifying polioviruses within countries; 16 regional reference laboratories, providing intratypic differentiation of wild and vaccine strains and assisting with quality assurance and training; and 6 global specialized laboratories, conducting research, preparing reference reagents, and providing genomic sequencing of wild polioviruses, advanced training, and expert virologic advice. Laboratories collaborate with national eradication programs in the detection, reporting, clinical investigation, and virologic testing of stool specimens obtained in connection with cases of acute flaccid paralysis and, where indicated, from healthy children and the environment. A quality assurance system, leading to World Health Organization accreditation, involves training in standardized techniques, use of centrally prepared typing antisera, annual proficiency testing and follow-up action, and monitoring of standard performance indicators. PMID- 9203703 TI - Development and coordination of the Polio Laboratory Network in the Western Pacific Region of the World Health Organization. AB - A multitiered network of polio laboratories, consisting of specialized reference laboratories, regional reference laboratories, national laboratories and, in the case of China, provincial laboratories, was established in the Western Pacific Region of the World Health Organization (WHO) in 1992. The network currently consists of 43 laboratories within the Region and is coordinated through the WHO Regional Office in Manila. As the levels and extent of supplementary immunization and acute flaccid paralysis surveillance activities have increased, so has the work load of network laboratories. The total number of stool specimens collected and processed in Polio Laboratory Network laboratories in this WHO region in 1995 exceeded 15,000. With the Region now establishing the criteria necessary for certification of polio-free status, it is essential for the Polio Laboratory Network to establish international confidence in its ability to carry out its role in the eradication of polio. PMID- 9203705 TI - A theoretical framework for evaluating the sensitivity of surveillance for detecting wild poliovirus: I. Factors affecting detection sensitivity in a person with acute flaccid paralysis. AB - Surveillance for cases of acute flaccid paralysis provides a means for detecting circulating wild poliovirus in a population and therefore is crucial to the global polio eradication effort. An initial step toward developing a more general framework for understanding the sensitivity of the acute flaccid paralysis surveillance system is presented by first specifying four categories of sensitivity involved: laboratory sensitivity, specimen sensitivity, person sensitivity, and population sensitivity. Using this framework, estimates for specimen sensitivity (the probability that virus will be detected in a specimen collected from an infected person) and the prevalence of infection are derived and applied to surveillance data from three regions. On the basis of the framework, our analysis indicates that a second specimen may significantly increase person sensitivity under some circumstances but provides little improvement under others. PMID- 9203704 TI - Surveillance for polio eradication in the People's Republic of China. AB - A case-based virus surveillance system for wild poliovirus in China was developed. By 1993, all 30 provincial immunization units and, by 1994, all 29 provincial laboratories were sending computerized data to the national level. In 1993, a county-level, computerized map was operationalized that permitted visual monitoring of the progress of the polio eradication program every month by county. In 1993, wild poliovirus type 1 was detected in 8 provinces. Wild poliovirus mainly caused clusters of polio cases identified by a surveillance system that detected primarily clinical polio in children <5 years old (1 stool sample was collected on approximately 50% of reported cases). By 1995, the surveillance system had reached certification-like levels (80% of acute flaccid paralysis [AFP] patients with 2 stool specimens and AFP case rate of 1/100,000 children <15 years old). No indigenous wild poliovirus was detected in 1995. This general case-based model can be applied to measles and other important diseases, and may then lead to a more rapid decrease in adverse health outcomes. PMID- 9203706 TI - A theoretical framework for evaluating the sensitivity of surveillance for detecting wild poliovirus: II. Factors affecting detection sensitivity in a population with circulating wild poliovirus. AB - A basic framework for describing sensitivity of surveillance to detect poliovirus is extended from individuals to general populations (population sensitivity). Using the mathematical formulations for population sensitivity, the theoretical behavior of surveillance sensitivity under various conditions is analyzed. As a region nears the elimination of poliovirus, population sensitivity falls to a value lower than the case-to-infection ratio, regardless of the system proficiency. Also, a second stool specimen makes a substantial contribution to population sensitivity only in regions with relatively low specimen sensitivity and then only over a narrow range of population infection incidence. Estimates of the mean numbers of infected and uninfected acute flaccid paralysis cases investigated in a season are derived. These may serve as additional indicators of system operation but require the collection of 2 specimens per case. PMID- 9203707 TI - Poliomyelitis surveillance: the compass for eradication. AB - Effective disease surveillance is a key strategy of the global polio eradication initiative. In an effort to strengthen the quality of polio surveillance as a prerequisite to achieving and certifying eradication, surveillance assessments were conducted in 28 countries in the World Health Organization African, Eastern Mediterranean, and European Regions from 1992 to 1995 using a standard protocol and evaluation guidelines. Six general recommendations were made: Use surveillance data for public health decision-making and action, improve timeliness of information exchange and dissemination, standardize the data collected, ensure adequate surveillance infrastructure, improve local data analysis, and enhance teamwork among surveillance partners. The experience gained will position the Expanded Programme on Immunization to address the challenges of disease prevention in the 21st century. PMID- 9203708 TI - Assessment of Guillain-Barre syndrome mortality and morbidity in the United States: implications for acute flaccid paralysis surveillance. AB - To estimate age-specific incidences and assess the national morbidity and mortality burden for Guillain-Barre syndrome (GBS) in the United States, a national hospital discharge database compiled by the Commission on Professional and Hospital Activities (CPHA) and national death certificate data reported to the National Vital Statistics System were reviewed. During 1985-1991, 10,453 patients with GBS were discharged from CPHA-participating hospitals (estimated annual incidence, 3.0/100,000 population). The age-specific incidence of GBS increased with age from 1.5/100,000 in persons <15 years old to 8.6/100,000 in persons 70-79 years old. The total estimated number of GBS-related deaths from 1985 through 1990 was 3770 (95% confidence interval, 3506-4034), for an average of 628 GBS deaths per year. These rates suggest that the proposed national surveillance system for acute flaccid paralysis should capture at a minimum the 796 GBS cases in persons <15 years old. GBS remains a significant health burden among older adults in the United States, with a marked increase in risk after age 40. PMID- 9203709 TI - Polio eradication: surveillance implications for the United Kingdom. AB - The requirements for certification of elimination of wild virus poliomyelitis will pose particular problems for some industrialized countries, such as the United Kingdom, where there has been no case detected for at least a decade. Systems of surveillance of poliomyelitis have been reviewed and potential weaknesses identified. When oral polio vaccine is routinely used, the rate of vaccine-associated cases provides an indication of the likelihood that if they occurred, wild virus cases would be detected. Acute flaccid paralysis surveillance was done for 3 years, but rates were lower than reported elsewhere and were accepted for certification purposes. Alternative techniques, such as surveillance of polioviruses, either in clinical samples or from the environment, may be developed in such countries. The ability to identify enteroviruses and to distinguish between wild and vaccine strains of polioviruses will give assurance that silent transmission of wild viruses is unlikely. PMID- 9203710 TI - Epidemiologic study of Guillain-Barre syndrome in children <15 years of age in Latin America. AB - In 1986, surveillance of acute flaccid paralysis (AFP) cases among children <15 years of age was implemented in Latin America as part of the initiative to eradicate poliomyelitis from the Western Hemisphere. Data on AFP, including Guillain-Barre syndrome (GBS), could be analyzed from a regional registry system and from specific GBS studies in seven countries. Between 1989 and 1991, 3112 cases of GBS were reported in Latin America, representing 52% of all nonpolio AFP cases. From the studies in seven countries, a total of 1527 GBS cases (49%) were studied, representing an overall annual incidence rate of 0.91/100,000 children <15 years old. Follow-up investigations showed a persistent muscular weakness at 60 days, 6 months, and 1 year after onset in 61%, 14%, and 10% of children, respectively. This study confirms that with the disappearance of polio, GBS arises as the most common cause of AFP. PMID- 9203711 TI - Outbreaks of paralytic poliomyelitis, 1976-1995. AB - During 1976-1995, 48 outbreaks of paralytic poliomyelitis with a cumulative total of approximately 17,000 cases were reported worldwide. Outbreaks occurred on most continents, affected from 0.1 to 52 persons per 100,000 total population (median, 4.4), lasted 2-25 months (median, 7), typically involved unvaccinated or inadequately vaccinated subgroups within highly immunized communities, and were primarily caused by poliovirus type 1 (74%). Cases in developing countries occurred predominantly among children <2 years of age, while those in industrialized countries tended to occur in older persons who had escaped natural infection earlier in life and who had not been vaccinated or had received poliovirus vaccine of inadequate potency. Partial genomic sequencing studies indicated that at least 15 outbreaks resulted from importation of wild polioviruses, primarily from the Indian subcontinent. These findings illustrate the potential for wide dissemination of wild poliovirus infection and underscore the critical need for maintaining high levels of immunity in all countries and for more aggressive vaccination efforts in areas in which polio is endemic. PMID- 9203712 TI - Establishing acute flaccid paralysis surveillance under difficult circumstances: lessons learned in Cambodia. AB - The implementation of the World Health Organization's recommended strategies for polio eradication, particularly acute flaccid paralysis (AFP) surveillance, can be limited by difficult circumstances beyond the control of immunization personnel. In Cambodia, however, obstacles to establishing AFP surveillance were rapidly overcome using a strategy that improved reporting through active surveillance in a geographically limited area before gradually expanding to include the whole country. The success of the strategy was ensured by the timely provision of the resources that were needed to establish, expand, and monitor surveillance activities. PMID- 9203713 TI - Duration of poliovirus excretion and its implications for acute flaccid paralysis surveillance: a review of the literature. AB - Timely investigation of children with acute flaccid paralysis, with collection of stool specimens for virus isolation, is the primary strategy used to detect wild poliovirus circulation. To determine the optimal timing of stool specimen collection, studies of wild and vaccine poliovirus excretion published between 1935 and 1995 were reviewed. Data were compiled from comparable studies, scatter plots of the data were created, and third-order regression lines were calculated. The data indicated that wild polioviruses were excreted by a majority of previously unvaccinated infants and young children for 3-4 weeks. The duration of viral shedding was reduced, however, among children who were previously vaccinated with inactivated poliovirus vaccine, who had preexisting antibodies to the infecting serotype, or who had previous intestinal infection with homologous poliovirus. These data suggest that the 14-day period after onset of paralysis is the interval with the highest probability of detecting wild poliovirus excretion in paralyzed children. PMID- 9203714 TI - National immunization days: state of the art. AB - National immunization days (NIDs) are nationwide mass campaigns to deliver supplemental doses of oral poliovirus vaccine to interrupt the circulation of wild polioviruses. They constitute one of the critical strategies for global poliomyelitis eradication and should be implemented in all countries with widespread poliovirus transmission. The certification of wild poliovirus eradication from the Western Hemisphere in September 1994 verified the effectiveness of this aspect of the World Health Organization's (WHO) overall strategy for polio eradication by the year 2000. NIDs require careful advanced planning and orchestration by each country. WHO provides specific guidelines for NIDs regarding the season, target age group, duration, frequency, inclusion of other interventions, vaccine delivery strategies, and evaluation. With strong routine immunization programs and the effective implementation of NIDs, "mop-up" campaigns, and acute flaccid paralysis surveillance, the goal of global polio eradication will be achieved. PMID- 9203715 TI - National immunization days: experience in Latin America. AB - Organization of national immunization days (NIDs) in all countries in Latin America in which polio was endemic has been one of the key elements that led to the interruption of the circulation of the wild poliovirus in 1991 from the Americas. National initiatives for control or elimination of measles using similar strategies have emerged from the successful organization of NIDs for polio eradication and lead the way to the eventual global eradication of this major killer of children. The major reasons for the success of polio eradication in the Americas were the commitment of national authorities, well-defined strategies, sustainable effort, and the participation of all sectors of society. PMID- 9203716 TI - The experience of countries in the Western Pacific Region in conducting national immunization days for poliomyelitis eradication. AB - Experience with national immunization days (NIDs) in six countries of the Western Pacific Region has shown that political support at all levels, detailed logistics plans, strategies appropriate to the local situation, and simple social mobilization messages have been key factors in the success of NIDs. Conventional strategies that may apply to conducting routine Expanded Programme on Immunization vaccinations do not necessarily apply to NIDs, in which the maximum number of children must be immunized in 1 or 2 days. Setting up temporary immunization posts at sites convenient to the local situation, moving the posts once or twice during the course of a day, and using volunteers to staff them are among many of the adaptations used successfully. Coverage figures published immediately after an NID can be misleading because of uncertainty about the true denominator. The true measure of the success of NIDs is in surveillance for wild poliovirus after the event. PMID- 9203718 TI - Evaluation of oral poliovirus vaccine delivery during the 1994 national immunization days in Pakistan. AB - Pakistan conducted national immunization days (NIDs) for the first time in 1994. To estimate coverage, to evaluate risk factors for failure to be immunized, and to determine the effectiveness of mass media, parents of 1288 children in 714 households in four districts were surveyed after the first NID round. In each district, a high proportion of children (93%-96%) received oral poliovirus vaccine (OPV) during the NID. In three districts, unimmunized or partially immunized children were less likely to receive NID OPV than were fully immunized children (Kohistan, P < .001; Quetta, P < .001; and Sibi, P = .05). Although a high proportion of children in each age cohort received NID OPV, in three districts children 0-11 months of age were less likely to receive NID OPV than were older children. Television and radio reached a high proportion of survey households, but other mass media were less effectively utilized. Risk factor and media effectiveness surveys provide important information that is useful for planning future NIDs. PMID- 9203717 TI - Increased immunogenicity of oral poliovirus vaccine administered in mass vaccination campaigns compared with the routine vaccination program in Jordan. AB - To compare the immunogenicity of routine versus mass campaign doses of oral poliovirus vaccine (OPV), serum neutralizing antibodies were measured in 254 children before and after two mass vaccination campaigns in Jordan. Precampaign seroprevalences to poliovirus types 1, 2, and 3 in children who had received three, four, or five routine doses of OPV were compared with postcampaign seroprevalences in children who had received one, two, or three routine doses plus two mass campaign doses. Seroprevalences were consistently higher in subgroups that received two doses through mass campaigns than in subgroups that received all doses through the routine program, especially for poliovirus type 3. Geometric mean titers were also consistently higher for mass campaign subgroups, particularly for poliovirus type 3. The findings suggest that adding further doses of OPV to the routine schedule is unlikely to have as great an impact on the immune state of children as administering the same number of doses during mass campaigns. PMID- 9203719 TI - Effect of target age of supplemental immunization campaigns on poliomyelitis occurrence in China. AB - The World Health Organization recommends conducting supplemental immunization activities to eradicate poliomyelitis by the year 2000. Although effective in eliminating poliomyelitis from the Americas, supplemental campaigns require substantial resources. To assess differential campaign effectiveness in eliminating this disease, poliomyelitis occurrence was compared in counties in China that targeted children <3 versus <4 years of age. Counties that targeted children <3 years of age reported poliomyelitis more frequently after the campaigns. This association was observed even after accounting for the effects of previous poliomyelitis occurrence, urban versus rural setting, and population density. While several limitations emphasize the preliminary nature of these findings, these data support targeting the widest possible age group of susceptible children to ensure maximum effectiveness in eliminating poliomyelitis. Thus, while reducing the target age of these activities may result in considerable resource savings, such campaigns may not be as effective in eliminating poliomyelitis. PMID- 9203720 TI - Combined immunization of infants with oral and inactivated poliovirus vaccines: results of a randomized trial in The Gambia, Oman, and Thailand. WHO Collaborative Study Group on Oral and Inactivated Poliovirus Vaccines. AB - To assess an immunization schedule combining oral (OPV) and inactivated poliovirus vaccines (IPV), a clinical trial was conducted in The Gambia, Oman, and Thailand. Children were randomized to receive OPV at birth and at 6, 10, and 14 weeks of age; OPV at birth followed by both OPV and IPV at 6, 10, and 14 weeks of age; or placebo at birth followed by IPV at 6, 10, and 14 weeks of age. Serum specimens were available at 24 weeks for 1291 (77%) of 1685 enrolled infants. In the combined-schedule group, the proportion of children seropositive at 24 weeks was 95%-99% for type 1, 99%- 100% for type 2, and 97%-100% for type 3. In The Gambia and Oman, the combined schedule performed significantly better than OPV for type 1 (95%-97% vs. 88%-90%) and type 3 (97%-99% vs. 72%-73%). Across the study sites, IPV given at 6, 10, and 14 weeks of age provided inadequate protection against poliovirus. The combined schedule provided the highest levels of serum antibody response, with mucosal immunity equivalent to that produced by OPV alone. PMID- 9203721 TI - Humoral and mucosal immunity in infants induced by three sequential inactivated poliovirus vaccine-live attenuated oral poliovirus vaccine immunization schedules. Baltimore Area Polio Vaccine Study Group. AB - The relative immunity induced by sequential administration of inactivated poliovirus vaccine (IPV) produced in human diploid cells and live attenuated oral poliovirus vaccine (OPV) was evaluated by randomization of 510 infants to receive IPV and OPV sequentially according to one of three experimental schedules, IPV only, or OPV only. The antibody response to two IPV doses was lower than expected. However, for each of the IPV-OPV sequential schedules, the first OPV dose significantly enhanced seroconversion rates and geometric mean microneutralization antibody titers. Three months after the final dose, 96%-99%, 99%-100%, and 81%-100% of infants had antibodies to poliovirus types 1, 2, and 3, respectively, and subjects with two or more prior OPV doses were significantly less likely than those with none or one prior OPV dose to excrete virus in feces after an OPV challenge. Sequential IPV-OPV immunization is now recommended for routine use in the United States. The optimal schedule consists of two IPV doses followed by two OPV doses. PMID- 9203722 TI - Sequential use of inactivated poliovirus vaccine followed by oral poliovirus vaccine in Oman. AB - Seroprevalence and geometric mean titers (GMTs) were compared at 6 and 10 months after vaccination with monovalent type 1 oral poliovirus vaccine (OPV) at 6 months and trivalent OPV at 7 and 9 months. Group 1 had received 4 doses of OPV, group 2 OPV at birth and 3 doses of OPV and inactivated poliovirus vaccine (IPV), and group 3 placebo at birth and 3 doses of IPV. A total of 547 infants completed the study. At 10 months, seroprevalence to poliovirus type 1 was 98%, 99%, and 98% in groups 1, 2, and 3; 100%, 100%, and 98% to poliovirus type 2; and 80%, 96%, and 91% to poliovirus type 3. Differences in seroprevalence among the groups were significant for poliovirus type 3 (P < .001). Between 6 and 10 months, significant increases in seroprevalence and GMTs occurred for poliovirus type 1 but not for types 2 and 3. Two OPV doses following 3 IPV doses did not significantly increase seroprevalence or raise GMTs for poliovirus types 2 and 3; however, significant increases were found for poliovirus type 1, which may have benefitted from monovalent type 1 administration. PMID- 9203723 TI - Sequential and combined use of inactivated and oral poliovirus vaccines: Dolj District, Romania, 1992-1994. AB - To determine the feasibility of a vaccination strategy that would reduce the risk of vaccine-associated paralysis while retaining a barrier against the spread of wild poliovirus, a 2-year project was undertaken using enhanced-potency inactivated poliovirus vaccine (IPV) administered at 2 and 3 months of age followed by doses of both IPV and oral poliovirus vaccine (OPV) administered at 4 and 9 months of age. Vaccination coverage by 12 months of age with three or more doses of IPV and two doses of OPV among 16,566 infants eligible for vaccination was > 95% and > 80%, respectively. Among 51 children from whom blood samples were obtained 45 days after their third dose of IPV and first dose of OPV, 100% had serum neutralizing antibodies (reciprocal titer > or = 10) to all three poliovirus types. No cases of paralytic poliomyelitis due to either wild or vaccine-related strains were reported. The project demonstrated the feasibility, safety, and high immunogenicity of sequential use of IPV followed by OPV in Romania. PMID- 9203724 TI - Development of a more thermostable poliovirus vaccine. AB - In 1991, a goal of improved thermostability of oral poliovirus vaccine (OPV) was set, and the Product Development Group was established under the Children's Vaccine Initiative to achieve this goal. Several initial research strategies were unsuccessful. The substitution of deuterium oxide for water in the final blending stage of vaccine production resulted in a significantly more stable product at temperatures of > or = 37 degrees C. A large body of clinical data shows the safety of deuterium at the dosage that would be given with this vaccine. However, reservations about the public acceptability of the vaccine, combined with the progress achieved in polio eradication with the current vaccine and the availability of vaccine vial monitors to indicate time and temperature exposure of every vial of OPV, have resulted in a recommendation that vaccine development cease. This product development activity has been instructive for the process of introduction of new vaccines. PMID- 9203725 TI - The clinical efficacy of trivalent oral polio vaccine in The Gambia by season of vaccine administration. AB - An epidemic of poliomyelitis caused by poliovirus type 1 occurred in The Gambia in 1986. To determine if a relationship existed between the failure of trivalent oral poliovirus vaccine (OPV) to prevent poliomyelitis and the season when children were vaccinated, 46 children 1-7 years old with poliomyelitis who had received three card-documented doses of OPV were compared with 260 controls who had also received three card-documented doses. Controls were individually matched with children who had poliomyelitis by age, sex, and residence. Children with poliomyelitis were more likely to have received doses in the rainy season (odds ratio describing the linear trend of each additional dose in the rainy season, 1.7; 95% confidence interval, 1.05-2.9). This finding extends previous observations of seasonal difference in the immunogenicity of OPV in The Gambia by showing that season of administration was associated with increased risk of vaccine failure nationwide for a several-year period. PMID- 9203726 TI - The effect of diarrhea on oral poliovirus vaccine failure in Brazil. AB - The effect of diarrhea on oral poliovirus vaccine (OPV) failure was evaluated using data from Brazil, where 728 infants were immunized at birth (OPV1) and approximately 6 (OPV2), 10 (OPV3), and 14 (OPV4) weeks. Recent diarrhea history was significantly associated with increased vaccine failure only after OPV2 for poliovirus types 2 and 3. In multivariate models, controlling for breast feeding, season of vaccine administration (type 3 only), maternal antibody (type 3 only), and immunization campaign exposure (type 3 only) strengthened this effect. Diarrhea at OPV receipt was associated with vaccine failure to poliovirus types 1 and 3 only after OPV2. These data support the current recommendation that children with diarrhea receive OPV and be reimmunized once their illness resolves. Expanding this recommendation to include children with a recent diarrhea history should be considered. While the effect of diarrhea on vaccine failure may be limited to OPV2, programmatic realities may preclude dose-specific recommendations. PMID- 9203728 TI - Strengthening routine immunization services in the Western Pacific through the eradication of poliomyelitis. AB - Infant immunization coverage in the Western Pacific Region of the World Health Organization was reviewed to evaluate the impact of polio eradication activities on routine immunization services. The trend in bacille Calmette-Guerin (one dose; BCG), diphtheria-tetanus toxoids-pertussis (three doses; DTP3), and measles (one dose) vaccination rates was analyzed from the beginning of eradication activities in 1990 to 1994 in the five polio-endemic countries that conducted supplementary oral polio vaccine immunization. In China and the Philippines, coverage for each antigen remained at or above 90% and 85%, respectively, while in Vietnam, coverage for all three antigens rose from 85% to 95%. BCG, DTP3, and measles vaccine coverage more than doubled in the People's Democratic Republic of Lao and increased by >30% in the Kingdom of Cambodia during the same period. PMID- 9203727 TI - Comparison of a monoclonal antibody-based IgM capture ELISA with a neutralization assay for assessing response to trivalent oral poliovirus vaccine. AB - Monoclone-based IgM capture ELISAs were developed for each of the three poliovirus serotypes and compared with a neutralization assay for detecting response to trivalent oral poliovirus vaccine among 224 infants. The IgM-based response rates were significantly higher than the neutralizing antibody-based rates: 95% versus 83% to poliovirus type 1, 99% versus 94% to poliovirus type 2, and 89% versus 59% to poliovirus type 3. IgM responses to the first vaccine dose were significantly associated between serotypes, suggesting that some of the discordance may reflect a heterotypic IgM response. When the response rates in 4 vaccine formulation groups were compared, group differences using the two assays were similar for poliovirus types 1 and 2 but not for type 3. Therefore, IgM results using these assays may not be adequate substitutes for neutralizing antibody results when determining vaccine response. PMID- 9203729 TI - Poliomyelitis eradication and its impact on primary health care in the Philippines. AB - Since 1992, the Philippines has conducted four national immunization days (NIDs) for polio eradication. Surveillance for acute flaccid paralysis (AFP) began in 1992. Through good routine immunization, the incidence of paralytic polio had decreased to low levels in the Philippines even before the NIDs were initiated. With continuously improving AFP and virologic surveillance, wild poliovirus has not been isolated since May 1993. NIDs had a direct positive effect on child health through supplementary immunization with oral poliovirus vaccine, measles vaccine, and tetanus toxoid, as well as through the distribution of vitamin A. Following the successful NIDs, the government budget for vaccine purchases increased significantly. Also, the NID strategy was used as a model for several other priority prevention programs of the Department of Health. Through the development of AFP surveillance, polio eradication also helped to improve surveillance for other Expanded Programme on Immunization diseases. PMID- 9203730 TI - Integrated disease control initiatives: polio eradication and neonatal tetanus elimination in Egypt. AB - Accelerated disease control initiatives, such as polio eradication by the year 2000, may substantially benefit public health programs in general. In Egypt, the control of other vaccine-preventable diseases, most noticeably neonatal tetanus (NT), has been facilitated by the polio eradication initiative. Linking NT reporting with the acute flaccid paralysis (AFP) surveillance system, which had been established for polio eradication, markedly improved the capacity to identify NT high-risk areas and target supplementary immunization activities appropriately. While the close integration of surveillance activities was to the benefit of both programs, mass immunization activities were not conducted simultaneously because of differences in the objectives, target populations, and operational aspects of oral polio vaccine and tetanus toxoid campaigns. In addition to substantial progress toward polio eradication in Egypt since 1988, there has been an 80% reduction in annual NT cases, in part due to the integration of appropriate aspects of these two disease control initiatives. PMID- 9203731 TI - Certification of the eradication of indigenous transmission of wild poliovirus in the Americas. AB - In May 1985, the Pan American Health Organization launched an initiative to interrupt indigenous transmission of the wild poliovirus from the Western Hemisphere by the year 1990. The strategy to achieve this goal was based on the maintenance of high levels of immunity in the population at risk and the establishment of a surveillance system to detect polio cases and respond promptly with control measures. On 23 August 1991, a 2-year-old boy with acute flaccid paralysis due to wild poliovirus was detected in Junin, Peru, the last isolation of such a virus in the entire Western Hemisphere. In 1990, an International Commission for the Certification of Eradication of Poliomyelitis Eradication (ICCPE) was established by the Pan American Health Organization to eventually determine if transmission was interrupted. After 3 years of follow-up and review of surveillance data, the ICCPE declared that wild poliovirus transmission had been interrupted in the Americas. PMID- 9203732 TI - The biologic principles of poliovirus eradication. AB - The biologic principles for the global eradication of poliomyelitis are as follows: Poliovirus causes acute, nonpersistent infections, virus is transmitted by infectious humans or their waste, survival of virus in the environment is finite, humans are the only reservoir, and immunization with polio vaccine interrupts virus transmission. These principles appear to be sound. The potential for prolonged virus excretion by immunocompromised patients requires further definition, although there is no epidemiologic evidence of a threat to eradication. Survival of poliovirus in the environment is highly variable, but viral inactivation is usually complete within months. Higher primates may be infected with poliovirus, but they are unlikely reservoirs in nature. The only poliovirus reservoir remaining after eradication will be laboratory stocks. Serious attention must be given to reducing this potential source of infection. Polio eradication through immunization is evidenced by the documented absence of poliomyelitis in an increasing number of countries and the progressive disappearance of poliovirus genotypes. PMID- 9203733 TI - [Sweet's syndrome. Presentation of six cases and review of the literature]. AB - The acute febrile neutrophilic dermatosis or Sweet syndrome, initially described in 1964 by Robert Sweet (1). It is characterized fever, neutrophilic leucocytosis, abrupt appearance of erythematous, painful, cutaneous plaquets and dense dermal infiltrate consisting of mature neutrophils without vasculitis sings. Malignancy has been described in the 10-15% of the reported cases. We report our series of 6 patients diagnosticated of this illness in our department. One of this patients has Sweet syndrome associated with a malignancy disorder. All of them had diagnostic criteria of the described disease and had good response to corticotherapy. We also report a bibliographic review of this infrequent syndrome. PMID- 9203734 TI - [Systemic capillary leak syndrome]. AB - The systemic capillary leak syndrome is a rare condition, with a high mortality rate, characterised by recurrent episodes of generalized edema, with hemoconcentration and hypoproteinemia, associated with paraproteinemia. Pathophysiologically, this syndrome is caused by a sudden, reversible increase in capillary permeability, with a rapid shift of plasma from the intravascular to the extravascular compartment with subsequent hypovolemic shock. We report a new patient of this unusual condition, and we review its diagnostic and therapeutic aspects. PMID- 9203735 TI - [Cat scratch disease and associated infections]. AB - Cat scratch disease (CSD) was first described in France by Debre et al. in 1950, yet the causative bacterial agent of CSD remained obscure until 1992, when Bartonella (formerly Rochalimaea) henselae was implicated in CSD by serological and microbiologic studies. B. henselae had been linked initially to bacillary angiomatosis (BA), but also bacillary peliosis, relapsing bacteremia and endocarditis. Cats are healthy carriers of B. henselae and B. clarridgeiae, and can be bacteremic for months to years. Cat to cat transmission of the organism involves the cat flea in absence of direct contact transmission. Present knowledge on the etiology, clinical features and epidemiological characteristics of cat scratch disease/bacillary angiomatosis are presented. PMID- 9203736 TI - [Urban atmospheric pollution: sources, pollutants and evolution]. AB - (Sub) urban air pollution (AP) is one of the most common and important forms of AP. This paper shortly gives some informations about the main sources of AP (mainly car traffic), the essential determinants of AP (emissions, meteorology, ..), the complexity of effluents and emissions (leading to the choice of AP indicators), the notion of exposition, and the recent evolution of environmental concentrations of air pollutants. Important progress were made towards emissions of industrial and heating sources, with a substantial decrease of SO2 and particulates air concentrations; but despite a marked improvement of fuels quality and of unitary vehicles emissions (and a correlated decrease in Pb and CO emissions) car pollution remains important in terms of photo-oxidant air pollution (CO, NOx, COV, O3); the good use of car in urban area must be reconsidered. PMID- 9203738 TI - [Urban atmospheric pollution and public health: epidemiologic data]. AB - Air pollution which is generated by car traffic of by various industries is susceptible to alter several functions of the respiratory cells. This has been demonstrated in several studies in vivo, in animal models or in human volunteers. However, the results of epidemiological studies in which the impact of air pollution on human health and particularly on the respiratory system (e.g asthma or respiratory infections), are unclear. Short term epidemiological studies have shown that air pollution can be responsible for some asthma symptoms such as cough and shortness of breath in known asthmatics, particularly in patients with mild rather than severe asthma. Moreover, these studies do not demonstrate than air pollution is responsible for the increase prevalence of asthma which has been reported these last 20 years. Concerning the relationships between air pollution and respiratory infections, the epidemiological studies are also contradictory. PMID- 9203737 TI - [Atmospheric pollution, mechanisms of action of inhaled toxicants]. AB - Inhaled pollutants have different targets in the airways. Ozone is able to induce lesions in all parts of the respiratory tract, SO2 is mainly solubilized in proximal airways while particles have a major retention time in the deep lung. Ciliated cells and pneumocytes 1 are the most sensitive targets. Besides the direct toxic effects of the pollutants on the target cells, amplification loops are easily switched on by indirect action of inflammatory mediators (Arachidonic acid cascade, PAF, cytokines, histamine, proteins secreted by eosinophils, proteases ... ); this is mainly due to the ability of injured epitheliums to recruit rapidly circulating inflammatory cells. Decreasing ventilatory function results mainly from triggering of nerve fibers which results in return in a true neurogenic inflammatory response. Chronical exposures to oxidants, sulfur derived compounds and particles may lead to changes of the homeostatic imbalance of airways cellularity. PMID- 9203740 TI - [Human life and energy production. Prospects opened up by controlled thermonuclear fusion]. AB - The massive and presently increasing energy production is going to confront mankind with a very important problem in the forthcoming decades, in particular due to the vanishing of resources and to the greenhouse effect. The share of fossil fuels in the energy production will have to decrease, and other energy sources will be needed. Among them controlled thermonuclear fusion has may assets due to its non-radioactive fuel with plentiful supply, its non radioactive and non polluting ashes, its safety, its weak environmental impact, and its irrelevance to nuclear proliferation in a normal setting. During the last three decades, physicists have made a series of steps toward the peaceful use of the dominant source of energy in the Universe. They have learned how to confine by magnetic fields plasmas at temperatures of 200 millions degrees centigrade, and they have developed several specific technologies. This way, they produced 11 million watts of nuclear power by fusing two isotopes of hydrogen. These investigations are conducted in a responsible spirit, that of ecoproduction, where possible negative consequences are anticipated, are made as low as reasonably achievable, and their management is studied. Yet several fundamental issues still have to be solved before on economically efficient industrial thermonuclear power plant be operated. A huge international collaboration involving Japan, the USA, the Russian Federation, and the European Union joined with Switzerland and Canada, is presently designing the first experimental thermonuclear reactor, the International Thermonuclear Experimental Reactor (ITER). It would cost 9 billion dollars, a cost similar to other large scientific projects. This is an important step toward an electricity producing thermonuclear reactor that would be both safe and respectful of human health and of environment. PMID- 9203739 TI - [Susceptible populations and urban atmospheric pollution. Development of a policy of protection]. AB - The short term vulnerability of sections of the population to the harmful effects of chemical pollutants in the urban atmosphere represents an important unresolved problem of public health. These groups include young children, the elderly, asthmatic patients, and patients with chronic bronchitis and cardio-respiratory insufficiency. A recently ratified French law on air and rational energy use is aimed at resolving this problem in the context of recognizing that everybody has a right to breathe air which is not damaging to their health. In order to achieve this objective and direct the measures to be taken to reduce the emission of pollutants relating to industrial activity, to heating and to road traffic, the law provides for an extension to the chain of automatic registering stations which enable the recording of atmospheric data in large urban centers. It also defines the limits of substances allowed in the air over given periods and revises the critical levels of particulate and gaseous pollutants allowed according to current experimental and epidemiological data. Thresholds for information and for alerting the population will also be defined, with eventually restrictive measures for industry and for car traffic envisaged. These provisions are a major step for the protection of susceptible populations and should be enhanced in the near future by extension to cover other potentially noxious substances. This requires continued study on the short and long term effects of atmospheric pollution on people's health. PMID- 9203741 TI - [The rehabilitation of chronic respiratory insufficiency]. AB - Respiratory rehabilitation is defined as a medical practice including a multidisciplinary medical program fitting each individual. Personalized retraining by means of exercises, is the master part of it, its aim is to improve the physical fitness in specialised institution then to maintain it when he becomes an out patient. In both cases, this retraining complies with strict rules concerning the mode of exercises (imposed power--duration of sessions--weekly frequency--progressiveness of overloading ...). This codification rests mainly on the recommendations of the American College of Sports Medicine. The choice of intensity at the beginning of the stay will be determined either by the maximal reserve of cardiac frequency or by the ventilatory threshold. This training has to involve extensive muscular mass and must not neglect the upper limbs. Ventilatory physiotherapy also plays an important part. The other components of rehabilitation concern optimisation of bronchodilator treatment, cessation of smoking, health education, physical education and relaxation, appraisal of nutritional status, assessment of therapeutic programs, of the quality of life and a long-term program for reinforcement of acquisitions. The therapeutic programs improve ventilatory performance, maximal oxygen intake, maximal tolerated power and quality of life. An adaptation of the St. George's Respiratory Questionnaire to patients hosted at the TOKI EDER Medical Center points out that the quality of life of patients with chronic respiratory failure is improved very highly significantly by this rehabilitation. PMID- 9203742 TI - [Attitudes and practices of drug users confronted with the risks of contamination by human immunodeficiency virus (HIV) and hepatitis B and C viruses]. AB - This study has been conducted in Lille, Marseille, Metz, Paris and its suburbs using an ethnographic methodology. A total number of 1,703 subjects using injectable drugs have been interviewed in the streets and in treatments centers. The result of the study are as follows: sharing syringes did progressively decrease since 1988; reusing syringes and sharing of the injection equipment maintained itself during the same time period; a decrease of HIV seroprevalence; a massive HVC contamination; qualitative and quantitative data do suggest the important role played by the multiple use of the same syringes and the sharing of the injection equipment. PMID- 9203743 TI - [The health of prisoners]. AB - In twenty years, the prison population, from the mother country and the overseas departments, has more than doubled, in spite of reprieve and amnesty decisions. This increase is more a consequence of longer penalties than of a rise in the number of imprisoned people The law no. 94-43, dated january 18th 1994, concerning the prisoners' medical care and welfare is an unprecedented health revolution. It comes in addition to provisions from 1986 and 1987 for the programme "13,000" prisons and those endowed with a regional medical and psychological service (SMPR). The prisoners' health must urgently be dealt with and particularly as regards infectious diseases, vaccination check-up, campaign against drug addiction health and nutrition education and dental care. As soon as incarceration has begun, the exist must be prepared and taken into consideration by the different interveners inside and outside the prison, in order to make sure of an efficient medical follow-up. As the number of intervening medical and social personnel, is increasing in prisons, a coordination inside the their walls as well as on the regional and national levels, would prove useful. PMID- 9203744 TI - Immunomorphological characteristics of pleomorphic adenoma of salivary glands. AB - The immunohistochemical profile of 23 pleomorphic adenomas and 7 normal salivary glands was studied. We used antisera to vimentin (V), desmin (D), epithelial membrane antigen (EMA), prostate specific antigen (PSA), pancytokeratin, carcinoembryonic antigen (CEA), glial fibrillary acidic protein (GFAP) and S-100 protein. In the ducts and myoepithelial cells of normal salivary glands immunopositivity to most of the cytoskeletal proteins, EMA and CEA was observed. GFAP was localized only in cells of striated ducts. Major differences in the expression of various antigens among tubular structures, solid sheets, the myxoid and chondroid in the pleomorphic adenoma were encountered. Appearance of GFAP as a sign of stromal transformation into myxoid and chondroid was detected. Judging from these comparative immunohistochemical characteristics between normal salivary glands and pleomorphic adenomas, we assume that tumour cells originate from the reserve cells of intercalated and striated ducts. PMID- 9203745 TI - Dental findings in patients with liver cirrhosis. A study of 100 cases. AB - A study was made of the dental findings in 100 patients with liver cirrhosis (LC) by examining the number of carious, missing and filled teeth. A significantly greater number of carious and missing teeth were observed in the patients with cirrhosis than in a control group of 50 healthy individuals. In the LC group, caries were found to affect more teeth in those patients with alcohol-induced LC than in those with liver disease of other causes. Finally, no relationship was observed between the number of carious, missing or filled teeth and certain determinations including serum glutamate pyruvate transaminase (SGPT), serum glutamate oxalacetate transaminase (SGOT), alkaline phosphate, platelet number, hepatitis B and C positivity markers, or antinuclear (ANA), antimitochondrial (AMA) or anti-smooth muscle autoantibodies (ASm). PMID- 9203746 TI - Localization of proliferating cell nuclear antigen-immunoreactivity in human dental pulp and gingiva. AB - The proliferating cell nuclear antigen (PCNA) is regarded as an operational marker of proliferating cells. We have used PC10 monoclonal antibody to PCNA to reveal proliferation sites in human dental pulp and gingiva. Intense PCNA immunoreactivity was observed in the basal layer of the gingiva lining epithelium and within some cells of the underlying connective tissues, including some endothelial and perivascular cells. PCNA-reactive cells were scattered throughout the pulp tissue, but were particularly numerous in the peripheral part. Since PCNA is an endogenous cell cycle-related molecule, we propose that PCNA antibodies may represent useful for studying cell kinetics in human oral tissues in normal as well as pathological situations, such as tumours, wound healing and inflammation. PMID- 9203748 TI - Managing managed care. PMID- 9203747 TI - Dental health in patients infected by human immunodeficiency virus (HIV). A study of 94 cases. AB - The dental health of 94 HIV-positive patients was investigated by the sum of carious, missing and filling teeth (DMF index), plaque (Silness and Loe) and bleeding point indices (Lenox and Kopczyk). Blood parameters were evaluated, including total lymphocytes, CD4+ lymphocytes, beta-2-microglobulin, platelets, the Quick index, and immunoglobulins (IgG, IgM and IgA). Patient oral parameters were contrasted with risk group (intravenous drug users homo- and heterosexuals) and controls. An evaluation was also made of differences in dental condition as a function of CD4+ cell number, according to the latest CDC classification (1992). HIV-positive intravenous drug users were found to present poorer DMF, plaque and bleeding point indices than the rest of the patients. No relationship was observed between CD4+ cell numbers in blood and dental condition, thus suggesting that the deteriorated dental health observed is a consequence of factors inherent to the risk groups involved rather than of the altered blood parameters. PMID- 9203749 TI - Exercise stress echocardiography; an alternative to perfusion imaging. PMID- 9203750 TI - Initial 100 consecutive stereotactic core breast biopsies in a private breast center setting. AB - BACKGROUND: Percutaneous stereotactic core biopsy has become an alternative to open surgical biopsy. STUDY: We report the initial 100 consecutive stereotactic core breast biopsies performed in a private breast center by three surgeons. RESULTS: Of the lesions biopsied, 89 were benign and 11 were malignant. Histological agreement was found in ten malignant and eight benign lesions that subsequently underwent open biopsy. In one patient with a lesion read as sclerosing papilloma on stereotactic core biopsy, a subsequent open biopsy undertaken at the pathologist's urging yielded a low-grade ductal carcinoma in situ. CONCLUSIONS: Stereotactic core breast biopsy is safe, accurate, cost effective, and minimally invasive. It will, in all likelihood, become the breast biopsy method of choice. PMID- 9203752 TI - How much should physicians worry about antitrust? PMID- 9203751 TI - Complex issues. PMID- 9203753 TI - Everyday problem solving in elderly women: contributions of residence, perceived control, and age. AB - This study examined the use of relativistic operations in everyday problem solving by elderly women. Thirty-two community residents and 32 nursing hostel residents aged between 65-90 years participated. Structured interviews canvassed the role of residence and other factors thought to contribute to the maintenance of problem-solving skills. Path analysis was used to test the developed model of the influences of age, engagement in problem-solving activities, control orientation, problem familiarity and residence on the use of relativistic operations. Partial support was found for the proposed model. Community residents used significantly more relativistic operations to solve the problems than did hostel residents. A revised model showed that in addition to residence and control orientation, the use of relativistic operations was influenced indirectly by the level of everyday problem-solving activity, through the orientation of perceived control. Increased problem-solving activity was associated with an internal control orientation, which in turn was directly related to the use of relativistic operations. The influence of age on problem solving was indirect, through problem-solving activity and residence. Our findings provide initial evidence of the extent to which variables other than age can influence everyday problem-solving performance. PMID- 9203754 TI - Measuring the adequacy of home care for frail elders. AB - Community elders (N = 330) were interviewed by telephone twice at baseline using both computer-assisted telephone interviews (CATI) and nurse clinicians, and at four-month follow-up using CATI to collect alternative paraprofessional home care adequacy measures. Measurement format influenced both reliability and observed levels of adequacy but alternative adequacy measures converged and were not closely linked to functional status. The predictive validity of measures based on clinical standards with respect to health outcomes was also demonstrated. These findings support emerging approaches in primary care and home health quality measurement that consider multiple dimension of user response to formal and informal care. PMID- 9203755 TI - Long-term determinants of patterns of health insurance coverage in the Medicare population. AB - Using data from the 1990 Health Supplement to the Panel Study of Income Dynamics, we examine the determinants of patterns of insurance coverage among the elderly. Among those with supplemental insurance through an employment-based source, the primary determinant of having insurance is work history, specifically job tenure and occupation of household heads and their spouses. Among those who do not have employer-provided insurance, wealth is the most important economic factor in the purchase of private insurance. Blacks, persons with less education and women household heads are less likely to purchase supplemental insurance. We find little evidence that persons in prior poor health are more likely to purchase supplemental insurance, and the most important determinant of dental or drug coverage is having employer-based insurance. The current trend toward decreased generosity of post-retirement benefits implies that fewer older Americans will have insurance for these services. PMID- 9203756 TI - Outpatient geriatric evaluation and management: is there an investment effect? AB - The effectiveness and efficiency of outpatient geriatric evaluation and management (GEM) was compared to usual outpatient primary care (UPC). Although GEM had no overall impact on health care utilization or cost of care for the entire study period, significant reductions were found during the sixteen- to twenty-four-month study period, suggesting a possible investment effect. In the first eight months of the study, GEM patients incurred 34.8% more in health care costs than UPC patients, but in the final eight months of the study the cost of care for UPC patients exceeded that for GEM patients by 37.8%. PMID- 9203757 TI - Legislating community care: the British experience, with U.S. comparisons. AB - In 1990, Great Britain enacted the National Health Service and Community Care Act to reduce unnecessary institutionalization, improve coordination of community based long-term care, and stimulate private service provision. An overview of four major principles of the legislation is provided: the elevation of the assessor/care manager role, the emphasis on inter-agency collaboration across the health and social care divide, privatization of service, and support of carers (caregivers). Preliminary findings related to their implementation indicate incremental rather than sweeping change. The elements of the new policy that borrow from American models of community care--and may be difficult to transplant -are highlighted. PMID- 9203758 TI - The influence of ethnicity and culture on the caregiver stress and coping process: a sociocultural review and analysis. AB - The authors review the literature on ethnic minority caregivers and suggest that ethnicity and culture play a significant role in the stress and coping process for Latino caregivers. Caregivers of older Latinos face special challenges in the caregiving for individuals at higher risk for specific chronic diseases, who are disabled at earlier ages, and who have more functional disabilities. Ethnicity and culture can also influence the appraisal of stress events, the perception and use of family support, and coping behaviors. Socioeconomic class and minority group status are discussed as additional sources of variation in the caregiver stress and coping model. PMID- 9203760 TI - The use of technical aids by elderly persons in The Netherlands: an application of the Andersen and Newman model. AB - In this article, variables were identified that are able to explain the use of technical aids by elderly people. The model developed by Andersen and Newman (1973) formed the basis for this research. Data were gathered from 498 randomly selected elderly people who were single, 75 years or older, and living independently. Whether or not elderly people use mobility aids or technical aids for basic activities of daily living (ADLs) is mainly predicted by need (functional status, chronic illnesses) and predisposing (gender, housing) variables. The number of used technical aids can be explained by predisposing (age, housing, education), enabling (income, receiving help), and need (functional status, chronic illness) variables. As opposed to others, in this study elderly people with a high income were less likely to use many mobility aids than people with an average income. We could not offer a plausible explanation for the nonlinear relationship between income and the number of technical aids used. PMID- 9203759 TI - Racial, ethnic, and cultural differences in dementia caregiving: review and analysis. AB - This study provides a review and analysis of the empirical research published since 1985 that has examined the impact of race, culture, and/or ethnicity on the dementia caregiving experience. Ten of the 12 studies included in the review focused on comparisons between Black and White caregivers; one examined differences between Black and Hispanic caregivers, and one focused on White and Hispanic caregivers. Compared to White caregivers, non-White caregivers: a) were less likely to be a spouse and more likely to be an adult child, friend, or other family member, b) reported lower levels of caregiver stress, burden, and depression, c) endorsed more strongly held beliefs about filial support, and d) were more likely to use prayer, faith, or religion as coping mechanisms. Strategies for advancing research in this area are discussed. PMID- 9203761 TI - Comparison of health and functional ability between noninstitutionalized and least dependent institutionalized elderly in Finland. AB - Finland's active deinstitutionalization policy aims to reduce the number of elderly people in long-term residential care and to keep noninstitutionalized elderly people living at home as long as possible. As a contribution to the issue of the appropriateness of long-term institutional care, we compared the health and functional ability of elderly people living at home or in residential care to assess the theoretical possibility of discharging the least dependent elderly from residential homes. Findings from two separate data sets collected in 1992 were compared; one (n = 475) was obtained by computer-assisted telephone interview (elderly at home) and the other (n = 459) by postal survey (elderly in residential care). The direct method was used in age and gender standardization, and logistic regression analysis was applied. Elderly people living at home were found to be in better health and with better functional ability than those in residential care. However, a proportion of home-dwellers needing some help with Activities of Daily Living (ADLs) assessed their health as being even worse than those in care, and approximated that of institutionalized elderly judged by the personnel to be able to manage with home-based care. Compared with home-dwellers, those assessed as able to manage in-home care were mostly single and had less education and more restrictions in their Instrumental ADLs and medication. Our results indicate that one third of those assessed as able to manage in-home care could possibly be discharged if adequate servicers and housing were available. PMID- 9203762 TI - The pattern of change in leisure activity behavior among older adults with arthritis. AB - This study employed a "ceasing participation" framework to examine changing leisure activity patterns. Respondents of the Living With Arthritis project were classified into four participation pattern categories. Results confirmed that older adults with arthritis are more likely to experience changes to their activity regimen than older adults without arthritis. A multi-group discriminant function analysis showed that arthritis severity distinguished those who tend to cease activity. Social network and age best distinguished those who quit activities without replacement. Results are placed in the context of coping strategies. Those who do not replace forfeited activities with other activities are least flexible in their response to their chronic condition and face challenges to their well-being. PMID- 9203763 TI - The developmental stake hypothesis and changing perceptions of intergenerational relations, 1971-1985. AB - This study examines the developmental stake hypothesis as it relates to changing perceptions of intergenerational relations among members of multigenerational families. The data come from the first two waves (1971 and 1985) of a longitudinal study of multigenerational family members (N = 1,057). We find strong support for the developmental stake hypothesis since, at both time periods, parents (both G1s and G2s) saw less distance in their relations with their children (both G2s and G3s). In addition, all respondents perceived less distance between the G2-G3 generations over time. We also found several gender differences in perceptions of family ties. Contrary to our expectations, only G1 respondents perceived less distance between generations in society in 1985 as compared to 1971. PMID- 9203765 TI - Children's views on aging: their attitudes and values. AB - Children's perceptions and attitudes about aging and older adults are investigated using a version of Children's Views on Aging (CVoA), a four-part validated instrument designed to assess school-age children's views on older adults and aging. The instrument has been adapted to enable children to make value judgements about their responses to questions on the CVoA. The study reports children's perceptions and attitudes about aging are not as negative as adults conclude. Children are positively affected by interactions with older adults, they describe physical signs of aging without judgement, and respond negatively to some of the unpleasant conditions associated with aging. PMID- 9203764 TI - A profile of grandparents raising grandchildren in the United States. AB - This article examines the prevalence of grandparent caregiving in the U.S. and presents a national profile of grandparent caregivers based on current data from the national Survey of Families and Households. More than one in ten grandparents are found to have cared for a grandchild for at least 6 months, with most of these having engaged in a far longer-term commitment. Although custodial grandparenting cuts across gender, class, and ethnic lines, single women, African Americans, and low income persons are disproportionately represented. Multivariate logistic analysis indicates that three groups--women, recently bereaved parents, and African Americans--have approximately twice the odds of becoming caregiving grandparents. Implications for further research, policy, and practice are discussed. PMID- 9203766 TI - Headache fear. AB - A prospective study of migraine patients seen at the office, over six months, was conducted to ascertain the prevalence and intensity of headache fear. With patients' help a questionnaire, with 14 questions and a maximum score of 50, was developed and refined. A total of 100 patients with episodic migraines (< 15 headache days/month) and another 100 patients with intractable migraines (> or = 15 headache days/month) were inducted. Fifty controls were selected from office patients who, on routine system review, were found to have migraine headaches but had never consulted a physician about them. The average fear score of the control group was 3/50 (range: 0 to 9), and only 2% felt a strong compulsion to take analgesics when headaches began. Based on that, headache fear was defined as a score of 10 or more. The average fear score of the episodic group was 12/50 (range: 0 to 43), 49% had a fear score of > or = 10, and 38% felt a strong compulsion to take analgesics. The average fear score of the intractable group was 19/50 (range: 0 to 46), 73% had a fear score of > or = 10, and 60% felt a strong compulsion to take analgesics. The average number of analgesic tablets taken per month in the control group was 7 (range: 0-150), in the episodic group 22 (range: 0-120), and in the intractable group 139 (range: 0-400). The three groups were comparable as far as age, gender, and migraine years. The frequency of migraines per month was 3 (range: 0-30) in the control group, 6 (range: 0-14) in the episodic group, and 29 (range: 15-30) in the intractable group. As empirically defined, headache fear is common and increases in a statistically significant manner across the three migraine groups (control-episodic intractable). I theorize that this fear may reinforce analgesic overuse and interfere with withdrawal efforts. Alloying this fear from the outset might provide patients with the support and reassurance needed for analgesic withdrawal, which is often necessary for recovery. PMID- 9203768 TI - It takes a revolution. Restoring the balance in health care. PMID- 9203767 TI - Herniated intervertebral disc without pain. AB - The diagnosis of herniated intervertebral disc is often made in cases of radicular pain in the low back, the neck, or sciatica or brachialgia. Practitioners often call upon radiologic imaging to confirm this diagnosis. But on radiologic examination, such a herniation may consist of a bulge, protrusion, prolapse, extension, extrusion or sequestration of this disc. We define and illustrate these terms from the literature. We then review the radiologic studies of normal controls, who have never had sciatica, brachialgia or pain in the low back or neck. In over one-quarter of these controls, studies using the plain x ray, CT scan, myelogram, and MRI show various radiologic signs of a herniated intervertebral disc. We therefore recommend that practitioners should not exclusively rely on radiologic imaging to confirm the clinical diagnosis of a herniated intervertebral disc. PMID- 9203769 TI - Report of the Ad Hoc Committee on the Oklahoma Health Care Authority and SoonerCare. PMID- 9203770 TI - A call for universal health coverage. PMID- 9203771 TI - Clarification of state HMO-Medicaid. PMID- 9203772 TI - Telemedicine: Oklahoma adopts strict new licensure rules for medical treatment employing electronic communication. PMID- 9203774 TI - Opportunity awaits for new and established dentists. PMID- 9203773 TI - Holistic dentistry. PMID- 9203775 TI - Clinical guidelines for indirect resin restorations. AB - Ongoing advances in adhesive dentistry have made it possible to successfully and predictably bond tooth-supporting restorations using conservative preparation techniques. Improvements in the durability and esthetic properties of tooth colored restorative materials have also increased the range of available treatment options. However, dentists have been slow to accept both direct and indirect posterior esthetics. This article provides a step-by-step technique for practitioners who choose to treat their patients with indirect resin esthetic restorations. It will not discuss other posterior restorative treatment techniques or materials (i.e. gold, porcelain, amalgam, bonded amalgam, or direct resin). PMID- 9203776 TI - Fifth generation bonding systems: state of the art in adhesive dentistry. PMID- 9203777 TI - Microleakage of cervical restorations etched with a weak organic acid. AB - This in-vitro study evaluated the microleakage of Class V restorations prepared using 10 per cent maleic acid and a composite resin. Thirty human premolar teeth were evenly distributed and randomly assigned to three groups. Conventional retentive preparations, etched with 10 per cent maleic acid for either 15, 30, or 60 seconds, were cut in the enamel on the facial surface of each tooth to a 1.5 mm depth (dentin). All teeth were restored with Z-100, a small particle composite resin. The teeth were then stored in deionized water for seven days, thermocycled, stained with methylene blue dye, invested, and sectioned vertically through the centre of the restoration. Leakage was established along each wall of the sectioned restoration. Analysis of variance (ANOVA) tests indicate that the restored teeth in Group 1 (15-second etch) had significantly greater microleakage (p < 0.05) on the enamel wall than the restored teeth in Group 2 (30-second etch) or Group 3 (60-second etch). In addition, Group 1 restorations had significantly greater overall microleakage (p < 0.05) than Group 2 or Group 3 restorations. Although the results were not statistically significant, it would appear that etching with 10 per cent maleic acid for 30 seconds could be clinically significant. Since a 30-second etch time was found to produce the least amount of microleakage (not statistically significant), it can be assumed that this etch time would also be optimal for etching enamel and dentin. Similarly, since Group 1 revealed the most overall microleakage, it can be assumed that a 15-second etch would be inadequate for etching enamel and dentin. Restorations in Groups 2 and 3 displayed statistically significant lower overall microleakage results. PMID- 9203778 TI - Effectiveness of salt versus glass bead sterilizers. AB - Microorganisms can be removed from dental instruments by various methods, including treatment in salt and glass bead sterilizers. However, no rigorous, controlled, in vivo or in vitro studies have been performed to verify the respective efficiencies of these methods. The goals of this study were to determine if the positioning of instruments at the centre or edge of a salt sterilizer results in differential sterilization effectiveness, and to compare the effectiveness of salt sterilizers relative to glass bead sterilizers. Autoclaved number 60 reamers were contaminated by plunging them to the handle in a commercial Bacillus stearothermophilus spore suspension. They were then sterilized for different periods of time and at different positions in the sterilizers. Each experiment included positive and negative controls. The results showed that better sterilization is achieved at the edge of the chamber than at the centre, and that salt sterilizers are more effective than glass bead sterilizers for a given period of time (15 seconds) in the sterilizer. PMID- 9203779 TI - The "ART" mandibular nerve block: a new approach to accomplishing regional anesthesia. AB - A new technique involving the use of a local block to anesthetize the inferior alveolar nerve (V3), a branch of the mandibular division of the trigeminal or fifth cranial nerve, is described. Clinicians fail to administer a successful mandibular block in as many as 15 per cent of all cases. This paper reviews and outlines some of the more common reasons for this failure, and how to avoid them. A short description of other techniques are presented, some of which should be reserved for isolated cases and not used on a routine basis. By avoiding or eliminating the reasons for mandibular block failure, and using the new block described in this paper, clinicians should be able to reduce the failure rate to much lower levels. PMID- 9203780 TI - The future of informed consent and patient-dentist communication. PMID- 9203782 TI - Holistic dentistry. PMID- 9203781 TI - Holistic dentistry. PMID- 9203783 TI - Holistic dentistry. PMID- 9203784 TI - Assignment of benefits. PMID- 9203785 TI - Gum protection. PMID- 9203786 TI - Erythromycin and trimethoprim-sulphamethoxazole in the treatment of cholera in children. AB - To evaluate the efficacy of erythromycin and trimethoprim-sulphamethoxazole (TMP SMX) in the treatment of cholera in children aged 1-8 years, a randomised clinical trial was conducted at a diarrhoea treatment centre in Bangladesh from December 1991 to June 1992. Fifteen children received erythromycin, 50 mg/kg per day, in four equally divided doses, 18 children received 10 mg/kg per day of trimethoprim and 50 mg/kg per day of sulphamethoxazole in two equally divided doses (12 hourly) for five days, and 15 children received no antibiotic; children in all three groups received intravenous cholera saline for severe dehydration and for mild to moderate dehydration, a rice-based oral rehydration solution. The mean stool volumes in mL/kg body weight in the two treatment groups were less than that of the control group, and there were no significant differences in stool volume among the two treatment groups. However, 67% of the children in the erythromycin group and 82% in the TMP-SMX group recovered within 72 hours compared to 33% in the control group (p < 0.01). Similarly, the bacteriological cures were 80% in the erythromycin group and 83% in the TMP-SMX group compared to only 27% in the control group (p < 0.001). These results confirm that both erythromycin and trimethoprim-sulphamethoxazole are effective antimicrobials in the treatment of cholera. These drugs are of value specially in younger children in whom tetracycline is contraindicated or when the infecting Vibrio cholerae are resistant to tetracycline. PMID- 9203787 TI - Differentiation of Vibrio cholerae O1 isolates with biochemical fingerprinting and comparison with ribotyping. AB - The Phene Plate (PhP) system is a commercially available typing system based on the measurements of kinetics of selected biochemical reactions of bacteria grown in liquid medium in 96-well microplates. The system uses numerical analysis to identify biochemical phenotypes among the tested strains. In the present study, a set of 16 discriminatory tests were used to differentiate 117 strains of Vibrio cholerae O1 from MExico and Bangladesh. The stability of PhP types of 16 isolates under different storage temperatures and after repeated subcultures were also evaluated. The PhP system had a reproducibility of 95%. Storage either at +4 degrees C or -70 degrees C, did not affect the reactions of the isolates, whereas 4 strains (25%) stored at room temperature and 5 strains (31%) subjected to 30 consecutive subcultures, exhibited minor changes in their biochemical reactions. Endemic isolates of V. cholerae O1 from Bangladesh were more diverse (diversity index = 0.84 to 0.93) than epidemic isolates from Mexico (diversity index = 0.73). Using a collection of 33 heterogeneous isolates of classical biotype of vibrios, PhP typing and ribotyping were compared. PhP typing discriminated more types (n = 23) than ribotyping (n = 5), whereas a combination of both yielded 27 types. The PhP system appears to be a simple, reliable and highly discriminating method for typing of V. cholerae, and may prove especially useful as a first screening method in epidemiological studies of V. cholerae. PMID- 9203788 TI - Adherence, invasion and cytotoxin assay of Campylobacter jejuni in HeLa and HEp-2 cells. AB - Campylobacter jejuni is an important human enteropathogen worldwide. Chickens are the major reservoir and source of campylobacter infection. Ten clinical isolates from human and five chicken strains were tested for the adherence, invasion and cytotoxin assay in HeLa and HEp-2 cells. All human strains adhered to both the HeLa (10(3) to 3 x 10(4) bacteria/mL of cell lysate) and HEp-2 cells (2 x 10(3) to 4 x 10(4) bacteria/mL of lysate). All chicken strains also adhered to the HEp 2 cells (10(2) to 10(3) bacteria/mL), but only two strains adhered to the HeLa cells. Six clinical and none of the chicken strains invaded the mammalian cells. Both the adherence and invasion were better observed in HEp-2 than in HeLa cell lines. All three isolates from patients having invasive diarrhoea and only one strain from a patient having watery diarrhoea produced cytotoxin. All three invasive strains also adhered to polystyrene surface after the localised destruction of the HEp-2 cells, a phenomenon not reported earlier. Adherence was markedly inhibited by the whole cell lysate and the acid glycine extracts, and the results were comparable. This study indicates that the clinical isolates of C. jejuni are more virulent than the chicken strains, HEp-2 is better for the adherence/invasion assay and HeLa is better for cytotoxin assay. The acid glycine extracts probably contain the key adhesins for C. jejuni. PMID- 9203789 TI - Mothers' health-seeking behaviour in acute diarrhoea in Tlaxcala, Mexico. AB - This study, a cross-sectional survey, was conducted to assess how mothers take care of their children with diarrhoea and to develop a model of health-care seeking behaviour. Multistage sampling was used. Mothers whose children aged less than five years had suffered from diarrhoea in the last fortnight were included. Nurses interviewed the mothers to collect data. Variables included in the interview were: mothers' characteristics, children's characteristics, clinical data, treatment given by the mother, maternal health-seeking behaviour and mothers' information about diarrhoea and dehydration. Variables corresponding to the clinical data were grouped to identify dehydration signs and the need for medical care. Dehydration was defined as the presence of two or more of the following reported signs: thirst, sunken eyes, sunken fontanelle, or scanty urine. The need for medical care was defined as the presence of one or more of the following characteristics: illness lasting more than three days, vomiting, fever, bloody diarrhoea or dehydration. A sample of 747 mothers was obtained. Household treatments consisted of herbal teas to stop diarrhoea (52.3%), liquids to prevent dehydration (92.2%), symptomatic drugs (35.2%) and changes in feeding patterns (36.3%), which consisted in suppressing milk and dairy products and interrupting breast feeding (12.2%). Mothers sought medical assistance when they perceived a worsening of clinical conditions. Clinical signs statistically associated with their decision were: bloody diarrhoea, vomiting, illness longer than three days, weight loss, and fever. The signs of dehydration were not associated with health care-seeking because the mother did not recognise them. It is concluded that maternal educational programmes should emphasise, besides the proper use of oral rehydration therapy, teaching mothers to identify signs of dehydration as an indication to seek timely medical care. PMID- 9203790 TI - Interaction between acute diarrhoea and falciparum malaria in Nigerian children. AB - Although both malaria and diarrhoea are major public health problems in developing countries, and separately each has been the subject of intense research, few studies have investigated the interaction between these two conditions. The interaction between diarrhoea and malaria among children aged 4 months to 12 years in two tertiary health-care facilities, University College Hospital, Ibadan, and Lagos University Teaching Hospital, Lagos, Nigeria was studied. In Ibadan, the prevalence of diarrhoea among the cerebral malaria patients on admission as 11.7% (7/60) compared to 9.3% (215/2312) among other admissions in 1990 (chi square = 0.16; p = 0.6913). Similarly, no significant difference in the prevalence of diarrhoea was found between the cerebral malaria patients (14.3%) and other patients (16.1%) seen in Lagos in 1992 (chi square = 0.06, p = 0.81). Thus, cerebral malaria does not seem to be associated with an increased or decreased prevalence of diarrhoea when compared with other conditions. The prevalence of malarial parasitaemia among the 554 diarrhoea patients studied in Ibadan during 1993-1994 was 13.6% compared with 17.9% among the 347 controls (chi square = 3.75, p = 0.053). However, of the children with diarrhoea, malarial parasitaemia was more common among the dehydrated patients (25.4%) than among the well-hydrated patients (11.6%) (chi square = 8.11, p = 0.004). These data suggest that diarrhoea is merely coincidental in severe malaria and conversely, malarial parasitaemia is similarly coincidental in children with acute diarrhoea, although it may be more frequent among dehydrated diarrhoea patients than well-hydrated ones. PMID- 9203791 TI - Production of haemolysin and enterotoxin by Aeromonas jandaei and Aeromonas trota strains after animal passage. AB - Five Aeromonas jandaei and 12 Aeromonas trota isolates were tested for the production of haemolysin and enterotoxin, and the correlation between these two properties. The majority (10 isolates) of the strains produced beta-haemolysis. The titres of haemolytic activity for both species were 8-64 HU/mL. In the initial ileal loop test, only two (A. trota) of the 17 isolates produced enterotoxin. One each of these 2 A. trota strains was beta-haemolytic and non haemolytic. The remaining isolates of A. trota and A. jandaei included alpha-, beta- and non-haemolytic strains, and failed to cause any fluid accumulation in the initial tests, but did so after one-to-five sequential passages through the rabbit ileal loops. Three alpha- and 4 non-haemolytic strains switched over to the production of beta-haemolysis when they showed the positive ileal loop reaction. However, on repeated subcultures or on storage in the laboratory, all of them reverted back to their original alpha- or non-haemolytic character and no longer produced enterotoxin. PMID- 9203792 TI - Bacterial lipopolysaccharide induces diarrhoea in caecectomized mice. AB - Castor oil, lipopolysaccharide of Escherichia coli, and endotoxin of Salmonella typhimurium were used for inducing diarrhoea in sham operated or caecectomized mice. Copious diarrhoea was induced by lipopolysaccharide (LPS) in caecectomized mice. Characteristics of diarrhoea induced by castor oil were not different between the two groups. It is concluded that caecectomized mice may be a good model to study lipopolysaccharide-induced diarrhoea. PMID- 9203793 TI - Antidiarrhoeal activities of Ocimum gratissimum (Lamiaceae). AB - The antidiarrhoeal activities of leaf extracts of Ocimum gratissimum were investigated by disc diffusion and tube dilution methods. The extracts were active against Aeromonas sobria, Escherichia coli, Plesiomonas shigelloides, Salmonella typhi, and Shigella dysenteriae. The leaf extracts were most active against S. dysenteriae and least active against S. typhi. The sensitivity of the organisms measured in terms of zone of inhibition ranged from 8.00 to 19.50 mm. The minimum inhibitory concentrations were from 4.00 to 50.00 mg ml-1, while the minimum bactericidal concentration ranged from 8.00 to 62 mg ml-1. The potentials of the leaf extract for the treatment of diarrhoeal diseases is discussed. PMID- 9203795 TI - Bibliography on diarrhoeal diseases. PMID- 9203794 TI - Epithelial adherence of Candida albicans is enhanced by passage through rat small intestine. AB - Seven Candida albicans isolates (four from patients with diarrhoea and three from healthy persons) underwent two passages through rat ileal loop (RIL) to see the effect of consecutive passages on the adherence to rat intestinal epithelium. The isolates from patients with diarrhoea showed a significant enhancement in adherence after the first passage (1.95 x 10(4) cfu/cm2 versus 3.67 x 10(4) cfu/cm2). There was no further increase between the first passage (3.67 x 10(4) cfu/cm2) and the second one (3.61 x 10(4) cfu/cm2). A similar pattern was observed with the three nondiarrhoeal isolates. Animal passage of this fungus probably leads to better interactions between the cell surfaces causing the enhanced adherence. PMID- 9203797 TI - The road best traveled. PMID- 9203796 TI - Easing performers' pain. PMID- 9203798 TI - Performing arts medicine crescendoes con spirito. PMID- 9203799 TI - A good walk spoiled. Golf's links to medicine. AB - This article explores the game of golf as it appears in the medical literature. Included are sections on the historical interaction of golf and medicine, the physiology and performance of golf, illness and injury related to golf, environmental concerns involving golf course management, physical disabilities, medical research and golf, and specific medical specialty references to golf. PMID- 9203800 TI - Shaken baby syndrome. Diagnosis and prevention. PMID- 9203801 TI - Geographic disparities in Medicare reimbursement. PMID- 9203802 TI - Are medical savings accounts an effective insurance alternative? PMID- 9203803 TI - Functional effects of lung volume reduction surgery. PMID- 9203804 TI - Utility of monitoring breathing during night hours in COPD patients undergoing long-term oxygen therapy. AB - We investigated the occurrence of nocturnal respiratory disorders during air and supplemental oxygen breathing in 16 patients with chronic obstructive pulmonary disease (COPD) undergoing long-term home oxygen therapy (LTOT). Following a first night of acclimatization, non-attended continuous nocturnal monitoring was performed for two successive nights in a randomized order. During one night patients breathed room air ("Air night"), and during the other they underwent LTOT ("O2 night") at the same protective O2 nasal flow rate set during waking hours in a resting state. O2 was administered from liquid reservoirs. On both occasions, the patients were monitored during the night for oxygen saturation by pulse finger oximetry (Sp,O2), chest-abdomen impedance, mouth-nasal thermistor flow rate, electrocardiogram (ECG), body position, eye movements, and leg movements. During the O2 night, compared to the Air night, mean (+/- SD) desaturation time decreased from 46 +/- 29 to 13 +/- 25%, while obstructive apnoea-hypopnoea duration increased from 6 +/- 8 to 9 +/- 7%, both expressed as percentage of total sleep time. The sleep apnoea/hypopnoea syndrome (SAHS) rate during the Air-night was 2 out of 16, both SAHS patients showing a reduction of apnoea-hypopnoea number.h-1 during the O2 night; whilst SAHS was noted in a further two patients during the O2 night. We conclude that Sp,O2 together with monitoring of breathing during the night, is potentially useful in patients with chronic obstructive pulmonary disease undergoing long-term oxygen therapy, not only when evaluating the O2 flow rate to be used during the night, but also for an understanding of the pathogenesis of nocturnal arterial oxyhaemoglobin desaturations, which may or may not be related to respiratory events. PMID- 9203805 TI - Inhaled beclomethasone dipropionate (BDP) prevents seasonal changes in atopic asthmatics. AB - Inhaled corticosteroids are most effective drugs currently available for the treatment of bronchial asthma. They have been shown to reduce airway inflammation and hyperresponsiveness. The aim of this study was to assess the preventive effect of inhaled steroid therapy in seasonal asthma. In a double-blind study, two groups of 10 allergic asthmatics were randomly assigned to receive inhaled beclomethasone dipropionate (BDP), 500 micrograms b.i.d., or a matched placebo, two puffs b.i.d. The patients used inhaled salbutamol as needed. At the beginning of the study, and every month between February and June, the following parameters were assessed: lung function (forced expiratory volume in one second (FEV1); airway responsiveness (provocative dose of methacholine producing a 20% fall in forced expiratory volume in one second (PD20)), serum eosinophil cationic protein (ECP); and blood eosinophil count. All subjects recorded daily asthma symptoms, beta 2-agonist consumption and peak expiratory flow (PEF) values. In the placebo group, all parameters except FEV1 worsened significantly during the pollen season compared with preseasonal values (p < 0.001). BDP produced complete protection, although a slight change from baseline was found for symptom score (p < 0.01), beta 2-agonist consumption (p < 0.01), and eosinophil number (p < 0.05) in May, when the pollen load was highest. These data provide evidence that beclomethasone dipropionate treatment is able to inhibit the seasonal changes occurring during natural exposure in asthmatics. This preventive effect is probably due to the anti-inflammatory action of beclamethasone dipropionate, as documented by its effect on serum markers of airway inflammation. PMID- 9203806 TI - Serum and pleural fluid levels of alpha-L-fucosidase and sialic acid have no diagnostic value in malignant pleural effusions. AB - The association of biological markers with cancer has been recognized for many decades. To determine whether alpha-L-fucosidase (ALF) and sialic acid (SA) are sensitive markers in malignant pleural effusions, they were investigated in serum and pleural fluid of 64 consecutive pleurisy patients, and in serum of 23 healthy subjects as a control group. The serum ALF (sALF) and serum SA (sSA) values of malignant and nonmalignant groups were higher than that of the control group, but the differences were statistically significant only for sSA determinations (p < 0.05). Values of sALF, sSA, pleural ALF (pALF), and pleural SA (pSA) were higher in the malignant group than the nonmalignant group, but no significant difference was found between the two groups. In conclusion, neither alpha-L-fucosidase nor sialic acid will be useful in the detection of malignant pleural effusions. PMID- 9203807 TI - The mortality rate of lung diseases eligible for transplant: national figures for 1989-1991. AB - Some patients with disabling and potentially fatal lung disease, who are nonresponders to conventional treatment, could benefit from lung transplantation (LT). It is, therefore, necessary to know the number of patients who are potentially eligible for such a therapeutic procedure. A retrospective study of the mortality rate of the Italian population from 1989-1991 was carried out. Groups of non-neoplastic chronic respiratory diseases, for which LT is indicated, were identified according to the Italian Central Statistical Institute (ISTAT), analytical international classification of diseases, trauma and causes of death (9th revision of 1975). The distribution of these diseases was considered both in terms of the total number of deaths (all ages) and in the relative number prior to 60 yrs of age (the maximum acceptable age for LT). Twenty five ISTAT codes referring to chronic non-neoplastic lung diseases for which LT is indicated were identified, and grouped according to disease type. The total national mortality rate from 1989-1991 due to selected lung diseases for which transplant is indicated was 44,915 (14,335 in 1989, 15,271 in 1990, and 15,309 in 1991), and 2,774 (6%) of these deaths occurred below the age limit of 60 yrs (986 in 1989, 889 in 1990, and 899 in 1981). Considering the normal limitations of retrospective studies on mortality rate, and the fact that only one eligibility criterion for LT (age) was considered, the results obtained provide an indirect evaluation and overestimation of the potential candidates for such treatment in Italy, and the relative need for organ donation. PMID- 9203808 TI - Lung transplantation: the experience of the Thoracic Organ Transplantation Centre of Pavia. AB - Between January 1991 and September 1995 at the Thoracic Organ Transplantation Centre of Pavia, 193 patients entered the waiting list for heart-lung or lung transplantation. Indications for heart-lung transplantation (HLT) were mainly primary or secondary pulmonary vascular diseases. Parenchymal lung diseases were the most frequent reasons for single- (SLT) or double-lung (DLT) transplantation. During the same period, 21 patients underwent HLT, 16 SLT and 14 DLT. Early deaths (within 30 days of surgery) occurred in 2 (10%) HLT, in 3 (19%) SLT, and in 3 (21%) DLT. Nineteen (90%) patients with HLT, 11 (69%) with SLT, and 10 (71%) with DLT survived up to 3 months; and 11 (52%) patients with HLT, 8 (50%) with SLT, and 5 (36%) with DLT survived up to 12 months. At the time of writing, the following patients are still alive: 10 (48%) with HLT, after a mean +/- SEM follow-up of 37.2 +/- 6 (range 28-46) months, 12 (75%) with SLT, after a mean follow-up of 16 +/- 11 (range 1-35) months, and finally 7 (50%) with DLT, after mean follow-up of 14 +/- 9 (range 1-23) months. PMID- 9203809 TI - Ineffectiveness of a four week treatment with the thromboxane synthetase inhibitor, imidazole salycilate, in reducing airway hyperresponsiveness to methacholine in asthmatics. AB - In a randomized, double-blind, placebo-controlled study, the acute and long-term effects of the reduction of thromboxane A2 (TxA2) synthesis on airway sensitivity and maximal airway narrowing in response to methacholine was evaluated in 12 subjects with mild-to-moderate stable asthma, using imidazole salycilate (IS), an anti-inflammatory drug which selectively inhibits the TxA2 synthetase. Dose response curves with methacholine (MCh) were performed in basal conditions (baseline); 1-1.5 h after administration of 1,500 mg of IS or placebo (acute); at 15 and 30 days of treatment with 750 mg t.i.d. of IS or placebo; and after a 2 week period of run-off (45 days). The serum levels of thromboxane B2 (TxB2) were measured at the same time points, except after acute administration, in five patients from each group. Baseline forced expiratory volume in one second (FEV1) was 78 +/- 7 and 85 +/- 8% of predicted in the IS and control group, respectively (NS). Throughout the study FEV1 remained unchanged in both groups, indicating that IS did not caused substantial modification of resting bronchial calibre. The initial provocative dose of methacholine causing a 20% fall in FEV1 (PD20) amounted to 27.0 +/- 1.5 micrograms in the IS group and 41.7 +/- 1.5 micrograms in the control group (geometric mean +/- GSEM) (NS). Despite a reduction of TxB2 serum levels with IS vs placebo at 15 days (24.9 +/- 8.5 vs 45.5 +/- 3.4 pg.mL-1; p < 0.05) and 30 days (27.0 +/- 6.3 vs 45.0 + 3.2 pg.mL-1; p < 0.05), MCh-induced bronchoconstriction, evaluated either as PD20 or maximal airway narrowing, did not change significantly during active treatment compared to placebo. These results show that prolonged reduction of thromboxane A2 synthesis does not improve airway sensitivity and limit maximal bronchoconstriction in asthmatic subjects, suggesting that thromboxane A2 per se does not play a substantial role in the pathogenesis of the airway hyperresponsiveness in human asthma. PMID- 9203811 TI - Severe pulmonary hypertension: a feature of Swyer-James syndrome? AB - Swyer-James syndrome, unilateral hyperlucent lung with air entrapment, generally occurs after severe infections during childhood. It is usually diagnosed by its characteristic chest radiographic image film or computed tomography, in patients who are almost asymptomatic. We report a case of Swyer-James syndrome, diagnosed from the study of severe pulmonary hypertension and with a fatal outcome. PMID- 9203810 TI - Pulmonary tumourlets and microcarcinoids in bronchiectasis. AB - A 66 year old woman underwent a left lower lobectomy for bronchiectasis. Histology revealed the presence of multiple endocrine lesions, such as neuroendocrine cell hyperplasia, tumourlets and microcarcinoids, which were widespread in bronchi, bronchioles and alveolar tissue. This case confirms the occurrence of neuroendocrine proliferations, in the setting of bronchiectasis, ranging from neuroendocrine cell hyperplasia to carcinoids, and suggests tumourlets as an appropriate model for neuroendocrine lung tumour genesis. PMID- 9203812 TI - The epidemiology of tuberculosis in Latvia. AB - An increase in the number of outbreaks of tuberculosis (TB) has been reported during recent years in many parts of the world, including the countries of Eastern Europe and the former USSR. This study was performed with the aim of assessing the current epidemiological situation with respect to TB in Latvia, one of the three Baltic States, in comparison to the previous decades (since 1950), and to evaluate trends in the incidence of TB during recent years. Data on the incidence of TB were obtained from the Tuberculosis Registers and the Information Department of the State TB and Lung Diseases Centre of Latvia. The results of this study show that, in Latvia, a deterioration in the epidemiological situation with respect to TB occurred from 1990 onwards. The incidence of TB and TB mortality rates virtually doubled from 1991 to 1995: from 28.7 to 50.4 and from 6.4 to 14.1 per 100,000 population, respectively. The majority of new TB cases occurred in persons from the economically productive age group, i.e. 25-54 yrs. In 1995, 61% of pulmonary TB patients were found to be sputum and/or culture positive, the rate of primary drug resistance was 19%, and of total drug resistance was 38%. In order to prevent a further deterioration in the epidemiology of TB in Latvia, it is vital to revise the National TB Control Programme according to the recommendations of the World Health Organization (WHO) and International Union Against Tuberculosis and Lung Disease (IUATLD). PMID- 9203813 TI - Assessment of the severity of pulmonary emphysema by computed tomography. AB - Pulmonary emphysema is defined as an abnormal enlargement of alveolar spaces distal to the terminal bronchioles, with alveolar wall disruption and without obvious fibrosis. Clinico-functional evaluation and chest radiographic diagnosis are not highly accurate in detecting emphysema and in establishing the extent of the process of alveolar destruction. Several computed tomography (CT) techniques are now available for detection and quantitative assessment of emphysema. The results appear to correlate significantly better than chest radiography with functional impairment and pathological score. Many options have been proposed by different authors regarding CT technique. The choice, however, is essentially between inspiratory high resolution CT (HRCT) with a visual scoring system, and automated quantitative evaluation by means of a "density mask" (DM) program. This paper presents the state of the art on CT quantification of pulmonary emphysema and briefly discusses the technical options and parameters to be used, together with the problems to be solved. PMID- 9203814 TI - Physiological outcomes of lung volume reduction surgery. AB - Lung volume reduction surgery (LVRS) is performed to alleviate dyspnoea of selected patients with severe pulmonary emphysema, and to improve their pulmonary function, performance in daily activity and quality of life. By resection of targeted emphysematous lung tissue, the achievable changes in pulmonary function consist of: 1) an increase in expiratory flow rate and airway conductance; 2) a reduction in hyperinflation accompanied by an augmentation in vital capacity; and 3) possibly, an improvement in gas exchange. Recent studies indicate that these changes are attributable to an increase of the lung's elastic recoil pressure. The consequences of an augmented recoil pressure consist of: 1) a reduction of pulmonary hyperinflation together with an amelioration of diaphragm and chest wall mechanics; 2) larger driving pressures; and 3) better airway stability. The combination of these factors is responsible for the improved pulmonary mechanics. PMID- 9203815 TI - Growth factors in asthma. AB - Asthma is a chronic inflammatory disease of the airways, with associated repair processes. Both inflammatory and repair processes appear to be strictly related, and can lead to several histopathological alterations of the bronchial mucosa, such as the shedding of epithelium and increased thickness of the basement membrane. The integrity as well as the alterations of the bronchial structure are the consequence of several biological events, such as cell proliferation and death, cell activation and inhibition, and extracellular matrix (ECM) production and degradation. These events are critically regulated by polypeptides called growth factors (GFs), which are able, functioning in an autocrine and paracrine fashion, to affect and modulate cell functions and ECM turnover. Although the importance of GFs has been widely demonstrated in other pulmonary conditions, such as lung fibrotic diseases, their possible involvement in the pathogenesis of inflammatory and postinflammatory processes in asthma is still not completely clear. The aim of the present review was to discuss the biological evidence concerning the role of several growth factors, such as transforming growth factor beta (TGF-beta), epidermal growth factor (EGF), granulocyte/macrophage colony stimulating factor (GM-CSF), platelet-derived growth factor (PDGF) and endothelin, in asthma and chronic bronchitis. PMID- 9203816 TI - Pulmonary failure as a cause of death in COPD. AB - Data on the outcome of patients with chronic obstructive pulmonary disease (COPD) are limited. We know that the prognosis is poor when respiratory insufficiency develops, but we have little information on the actual cause of death. Epidemiological studies are suitable for the assessment of the prevalence of the disease, but give no details on the actual cause of death. Age and forced expiratory volume in one second (FEV1) have been recognized as the best predictors of mortality in studies designed to quantify survival of COPD patients, particularly when the post-brochodilator value is used, as this provides a better estimate of airway and parenchymal damage. Data from Intensive Care Units on acute respiratory failure have several significant limitations. Firstly, it is probable that some patients elect not to undergo intensive treatment for a terminal bout of respiratory failure, particularly if it is not first episode. Secondly, the actual cause of death is often not described in adequate detail. Hypoxaemia and acidaemia are the main risk factors in acute exacerbation of the disease and the presence of pulmonary infiltrates on chest radiographs worsens the prognosis. A single bout of respiratory failure appears to have no effect on the prognosis of COPD patients after recovery, but there is a consistent increase in mortality after the second episode. It seems possible to manage the majority of episodes of acute respiratory failure with mechanical ventilation administered with noninvasive techniques. When endotracheal intubation is necessary, the prognosis is usually poor and the survival after 1 yr is usually lower than 40%. The role of long-term home mechanical ventilation is still unclear. Results from pivotal studies have been encouraging, although survival is far less impressive than in neuromuscular disorders. In patients with end-stage lung disease, lung transplantation can be considered the only possibility of increasing pulmonary functional capacity. However the technique is reserved only for a highly selected group of patients and data on the long-term outcome are awaited. PMID- 9203817 TI - Role for granulocyte/macrophage colony-stimulating factor in pulmonary surfactant homeostasis. PMID- 9203818 TI - Transcription factors as activators of gene transcription: AP-1 and NF-kappa B. AB - Cells respond to a range of cytokines and other inflammatory stimuli by selectively expressing a wide range of genes. These proinflammatory signals bind to receptors and initiate intracellular signalling cascades. This results in the activation of proinflammatory deoxyribonucleic acid (DNA)-binding proteins or transcription factors such as activator protein-1 (AP-1) or nuclear factor-kappa B (NF-kappa B). Following activation, these factors bind to specific recognition sequences in the control regions (promoters) of target genes causing modulation of gene transcription. Furthermore, numerous sites for regulation of cytokine and cytokine receptor genes by these transcription factors are found in their promoter regions. Many factors affect the formation and activity of AP-1 dimers (Fos and Jun heterodimers) through protein specific interactions or by the modulation of pre-existing complexes by phosphorylation. These complexes can vary markedly in their ability to stimulate gene transcription. Thus, induction of AP 1 activity is regulated by stimuli that either induce the de novo synthesis of AP 1 subunits or increase the activity of previously formed AP-1 dimers. NF-kappa B is activated by a number of agents including tumour necrosis factor-alpha (TNF alpha), interleukin-1 beta (IL-1 beta), lipopolysaccharide (LPS) and viruses. NF kappa B plays a central role in a range of immunological responses due to its ability to switch on inflammatory genes which lead to further activation of NF kappa B is a heterodimer of proteins which vary in their ability to bind to DNA and/or to activate gene transcription. NF-kappa B activation is primarily regulated by sequestration of heterodimers within the cytoplasm as inactive complexes with inhibitory molecules (inhibitory-kappa B (I-kappa B)). Treatment of cells with inducing agents results in the phosphorylation of the I-kappa B molecule which is targeted for rapid degradation. This causes a rapid dissociation of the cytoplasmic NF-kappa B/I-kappa B complexes and allows translocation of the active NF-kappa B to the nucleus. Cross-coupling between NF kappa B and AP-1 has been described which results in a synergistic increase in activity at both AP-1 and NF-kappa B sites. Elevation of both AP-1 and NF-kappa B, as reported in asthma, may therefore lead to far greater inflammation than would be present if either transcription factor alone were activated. PMID- 9203819 TI - Improved simulation system for routine cardiopulmonary exercise test equipment: introductory remarks. ECSC Working Group on Standardization Stress Test Methods. AB - The instruments used in the evaluation of cardiorespiratory function employ extraordinarily fast and complex techniques: instantaneous ventilatory flow volume transducer operate together with rapid CO2-O2 analysers, and are coupled to on-line cardiocirculatory monitors working in real time under the control of advanced software. It is clear that, with such powerful equipment, the calibrations currently made at rest before exercise are insufficient and must be accompanied by, or replaced with, methods capable of continuous (breath-to breath) monitoring of the instrument's activity throughout the exercise period. Two new simulation and check systems were developed for the calibration and the continuous monitoring of exercise equipment. The former applies to metabolic determinations, the latter to the cycle ergometers employed to impose varying workloads in the implementation of specific protocols. PMID- 9203820 TI - A new metabolic simulator system for routine cardiopulmonary exercise test equipment: technical specifications and validation. ECSC Working Group on Standardization Stress Test Methods. AB - Modern stress-test instruments require non-steady-state calibration (i.e., during exercise) instead of calibration at rest (i.e., before use) in stable conditions. The purpose of the present study was to describe a new specific computer controlled check system for stress-test instruments, which operates on the basis of physiological respiratory equations. It can produce rapid and automatic modifications of metabolic data, allowing breath-to-breath simulation. When connected to the stress-test equipment to be controlled, it makes it possible to identify discrepancies between preset and measured values. Systematic comparative measurements were performed using as reference two mass spectrometer-controlled analysers and a high-precision flow meter. Accuracy, trend and reproducibility in continuously changing conditions were very satisfactory. PMID- 9203821 TI - The PDA Training and Research Institute. PMID- 9203822 TI - A European perspective on regulation and technology. PMID- 9203823 TI - Investigation of pulsed light for terminal sterilization of WFI filled blow/fill/seal polyethylene containers. AB - A study was performed to assess the ability of pulsed light to sterilize water for injection in blow/fill/seal polyethylene containers. Pulsed light uses intense, short duration flashes of broad spectrum white light to produce high levels of microbial kill. In a first phase of testing, containers of 0.5, 5, 15, and 120 mL nominal volume were inoculated with Bacillus pumilus endospores, Bacillus subtilus variety niger strain globigii endospores, Bacillus stearothermophilus endospores, and Aspergillus niger conidiospores. Approximately 10(6) colony forming units of each test spore were individually inoculated into 22 replicate containers of each sample volume. Two of these containers served as inoculation recovery controls, and 10 were treated using each of two pulsed light exposure methods: single-sided treatment, or treatment within a reflective cavity. Both treatments employed flashes of intense broad spectrum pulsed light delivered at one flash per second. Cavity treatment used 10 flashes to treat each container within a reflective cavity containing a single lamp. Cavity treatment yielded no recoverable survivors for any of the challenge spores from the contents of any of the 160 total samples. Single-sided treatment used 20 approximately 1-J/cm2 flashes from a single lamp-reflector projecting onto one side of the container. Single-sided treatment yielded no recoverable survivors from the contents of the containers for any of the bacterial endospores tested, but Aspergillus niger survival was detected in 4 of the 40 single-side treated samples. A second phase of tests examined the pulsed light inactivation of Bacillus pumilus spores for a range of inoculation levels. High levels of Bacillus pumilus spore kill were obtained using only a few cavity flashes. The results show that pulsed light can provide high levels of microbial lethality and possesses potential for use as a terminal sterilization method for water for injection in filled, sealed polyethylene containers. PMID- 9203824 TI - Synergistic sterilization: a brief history. PMID- 9203825 TI - Virus retention by a hydrophilic triple-layer PVDF microporous membrane filter. AB - Retention of bacteriophages (phi 6, PR772, T1, and PP7) and mammalian viruses (poliovirus and influenza A virus) by a hydrophilic triple layer PVDF microporous membrane, the Ultipor VF grade DV50 membrane, was evaluated. Challenges of membrane discs or pleated filter cartridges were performed at concentrations of 10(6)-10(8) PFU/mL in one or more of the following carrier fluids: water, saline, gelatin (0.1%) in phosphate buffer, Dulbecco's Modified Eagle Medium (MEM), and MEM supplemented with 10% fetal bovine serum (MEM + 10). The data demonstrate a minimum log titer reduction (LTR) of 6 for viruses larger than 50 nm irrespective of the carrier fluid. Protein transmission levels of greater than 95% for IgG and albumin were achieved. For integral pleated filter cartridges, correlation between a nondestructive integrity test (using the forward flow integrity test method) and virus retention was demonstrated. The Ultipor VF grade DV50 filter can be applicable in the manufacture of biologicals and biopharmaceuticals, where high protein transmission and consistent viral titer reduction are desired. PMID- 9203826 TI - Effect of ultraviolet light on the release of theophylline from ethylcellulose based sustained-release microcapsules. AB - Sustained-release theophylline microcapsules were prepared using the emulsion solvent-evaporation technique. Three viscosity grades of ethylcellulose and varying concentrations of theophylline were used for the formulation. The microcapsules were exposed to short-wave ultraviolet (UV) light for 7 days. The surface and the cross-section of microcapsules were analyzed by scanning electron microscopy (SEM). The release of drug was determined by using the USP dissolution apparatus. Differential scanning calorimetry (DSC) was used to study the thermal properties of theophylline and the polymer. SEM revealed numerous pores on the surface of microcapsules. The size of the pores increased on exposure to light. Theophylline release from microcapsules was found to follow first-order kinetics both before and after UV light exposure. The drug release rate from microcapsules exposed to short-wave UV light increased by about 80%-450% compared with the unexposed samples. The release of theophylline from the microcapsules was more faster with higher concentration of ethylcellulose polymer. Both SEM and DSC showed uniform dispersion of drug with the polymer within the microcapsules. The melting point of the polymers changed, but it remained unchanged for theophylline on exposure to UV light. The UV light caused photodegradation or depolymerization of the polymer and thus caused a faster release of drug from the sustained release microcapsules. The finding underscores the need for setting standards and requirements for light-stability testing for sustained-release formulations, where there may be substantial change in the release pattern of drug on exposure to light. PMID- 9203827 TI - Dynamic compatibility testing of DMP 840, an experimental antitumor agent. AB - The purpose of this study is to evaluate the potential for DMP 840, a novel experimental antitumor agent, to precipitate during injection or dilution with infusion solutions. The influence of predilution of the drug solution before injection and addition of buffers to the drug vehicle were also investigated. The compatibility of normal saline solution, pH 7.4 phosphate buffers, and human plasma with DMP 840 was examined in vitro under both static conditions and dynamic flow. The combination of DMP 840 solutions with normal saline solution resulted in conversion of the drug to an insoluble dihydrochloride salt. Under conditions of dynamic flow, precipitation, accompanied by large changes in turbidity, occurred at relatively high concentrations of the drug in the injection solution. Dilution of the injection solution below 2 mg/mL or slow injection avoided precipitation. As was the case with the normal saline system, turbidity changes after injection into protein-phosphate buffer (PPB) were dependent on the initial concentration of DMP 840 solution as well as the rate of administration. In addition, the maximum injection rate at which complete miscibility occurred increased exponentially as the drug injection solution was made more dilute. Buffering the DMP 840 injection solution with acetate buffer improved the miscibility of DMP 840 with PPB, which indicated that the turbidity increases were most likely due to conversion of the drug to its insoluble free base form. The observed effects of the buffer on the turbidity response agreed qualitatively with predictions from a graphical approach that considers the effects of dilution and pH changes on drug solubility. Despite these observations, no evidence for the formation of a solid precipitate could be found after injection of the unbuffered drug solution into PPB. Further investigation indicated that the presence of albumin in the PPB prevented the formation of a solid phase during injection. Likewise, fresh human plasma, spiked with 1 and 2 mg/mL solutions of DMP 840, showed no evidence for the formation of a solid precipitate. PMID- 9203829 TI - Practice parameters: what are they and why should we use them? PMID- 9203828 TI - PDA comments: active pharmaceutical ingredients/guidance. PDA Parenteral Drug Association. PMID- 9203830 TI - A synopsis of the American Academy of Pediatrics' practice parameter on the management of acute gastroenteritis in young children. PMID- 9203832 TI - Giardia. PMID- 9203833 TI - Index of suspicion. Case 1 presentation. PMID- 9203831 TI - Infant growth and development. PMID- 9203834 TI - Index of suspicion. Case 2 presentation. PMID- 9203835 TI - Index of suspicion. Case 3 presentation. PMID- 9203836 TI - Are we ready to collaborate for community-based health services? PMID- 9203837 TI - The client-nurse relationship as experienced by public health nurses: toward better collaboration. AB - This article presents the findings of a study concerned with the client-nurse relationship in public health nursing settings. The purpose was to look at the ways the client and public health nurse cooperate and to see what makes for efficient collaboration. A phenomenological-hermeneutic approach was adopted. The data consisted of essays written by the public health nurses and focused interviews, which were analyzed by phenomenological method. The outcome is an interpretive description of public health nurses' experiences of collaboration with their clients. Successful collaboration requires an active and committed involvement on both sides and a joint effort to help the client cope with his/her situation. This means there has to be not only a shared understanding of the ultimate goal of nursing, but also open and sincere confidence-building interaction for the creation of a sense of confidentiality and trustworthiness. The results suggest that the contents of the client-nurse relationship are extremely important to both sides of the dyad: both the client's well-being and the public health nurse's feeling of succeeding on the job will depend to a great extent on the kind of relationship they construct. Future research should also look at how clients experience their relationship with the public health nurse. PMID- 9203838 TI - Enhancing client competence: melding professional and client knowledge in public health nursing practice. AB - Providing health information is an important aspect of public health nursing. This article describes how public health nurses (PHNs) give information to enhance client competence. The findings are part of a larger study that explored PHNs perspectives and experiences of their practice. The study employed an exploratory descriptive qualitative research design. Data were gathered through in-depth individual and focus group interviews with 28 PHNs in Alberta, Canada. Content analysis revealed that nurses work to enhance client competence by sharing professional knowledge and by building on the client's experiential knowledge. Nurses provide information to assist clients with immediate concerns and for future use, PHNs use three main strategies to deal with immediate concerns: being direct, providing options, and presenting a different view. Information for future use focused on enhancing development and forestalling future problems. Nurses build on clients' experiential knowledge by acknowledging their present situation, giving positive feedback, being there, and gently persuading. The authors suggest that the melding of professional and client knowledge is foundational to health promotion approaches that enhance client competence. There is a need for further research that explores the intricacies of developing partnerships between professionals and clients that embrace a sharing of professional and experiential knowledge. PMID- 9203839 TI - "Bridging the information gap" for Virginia public health nurses. AB - This project was designed to increase the public health nurse's knowledge and use of health science information resources available from the National Library of Medicine's databases through the use of the Grateful Med software program. In 1994, the Tompkins-McCaw Library located on the Medical College of Virginia Campus (MCV) of Virginia Commonwealth University (VCU) was awarded a Nursing Information Access Grant from the Southeastern/Atlantic Region of the National Network of Libraries of Medicine (NN/LM). This project was a collaboration of the Tompkins McCaw Library, the VCU School of Nursing, and The Virginia Department of Health. Sixty public health nurses received Grateful Med training. Session evaluations were conducted and indicate that although public health nurses received training and had access to health science information resources through Grateful Med, subsequent use of the resources was very limited. Similar to reports on information-seeking behaviors of physicians, public health nurses seek information from colleagues, personal collections, and other resources locally available. Reasons for the project's limited success in changing the health science information seeking and utilization practices of public health nurses are discussed, and potential solutions are proposed. PMID- 9203840 TI - Ethics in public health practice: a survey of public health nurses in southern Louisiana. AB - The present study was designed to help learn more about the ethical interests and concerns of public health nurses employed in state and local health departments. Self-administered postal questionnaires were mailed to 41 public health nurses employed at health units in Region I of the Louisiana Office of Public Health. Basic demographic information was obtained along with information about the workers' previous instruction or training in ethics and the nature of ethical conflicts encountered in their public health practice. Only 38% (15 of 39) of the surveyed nurses had had formal instruction in ethics. Even fewer (7.3%) had received continuing education on ethics. Most of the nurses felt confident in their ability to recognize an ethical conflict or dilemma in the workplace; fewer felt confident in their ability to resolve an ethical conflict or dilemma. A high proportion of the nurses agreed that there is a need for continuing education courses on ethics for public health workers. Nurses who had received formal ethics instruction were more likely to feel confident in their ability to recognize an ethical conflict in their public health practice. Continuing education programs on ethics are needed that are designed to meet the specific needs of front-line public health workers. PMID- 9203841 TI - Lead poisoning and associated risk factors among preschoolers enrolled in a Head Start program. AB - Despite the fact that lead poisoning is one of the most common pediatric health problems in the United States today, little is known about the prevalence and correlates of this disease among nonurban preschool children living in the southern United States. The purpose of this study was to measure the prevalence of abnormal lead levels and to explore the relationships between lead levels and gender, weight, hemoglobin, and ethnicity. Using a chart review protocol, data were collected from 81 charts of children enrolled in a Head Start program in Florida. The prevalence rate of elevated lead levels was 18.5%, a rate higher than that found in most previous research. No relationship was found between lead levels and gender, weight, hemoglobin, and ethnicity. The results highlight the importance of local screening efforts. Controversies in screening are discussed in this article in some detail with the aim of assisting health care providers make decisions about whether universal screening for lead levels in children is appropriate and whether use of the Centers for Disease Control questionnaire has sufficient value. Further study is needed regarding prevalence rates in different geographic areas in the United States, and factors associated with elevated lead levels. PMID- 9203842 TI - Caregivers' knowledge and perceptions of preventing childhood lead poisoning. AB - Philadelphia is considered a high-risk area for lead poisoning. The Philadelphia Department of Public Health conducts education and outreach to screen children and promote prevention. Prior to this study there were no systematic data on the community's knowledge and perceptions of lead poisoning prevention. A 32-item questionnaire was developed to assess knowledge and perceptions of family caregivers of children younger than age 8. The survey was interviewer administered in pediatric clinics in two geographic areas of the city with documented severe and moderate lead poisoning rates. Eighty family caregivers were interviewed while waiting to see the pediatrician. Caregivers in the sample were connected to a health care system and resided in areas where lead community outreach was concentrated. Nevertheless, results suggest that even caregivers of children in high-risk areas do not mention lead poisoning as a health concern. About 61% of the sample identified eating paint chips as a cause of lead poisoning, whereas only 15% identified lead paint dust as a source of lead poisoning. Approximately 49% of the caregivers reported that they "never" or only "sometimes" perform recommended prevention activities. The Philadelphia Department of Public Health used these findings to review and modify education and outreach to prevent lead poisoning. PMID- 9203843 TI - More does not mean better: prenatal visits and pregnancy outcome in the Hispanic population. AB - Early and consistent prenatal care (PNC) is thought to play an important role in the reduction of low birthweight (LBW) in the United States. It has been reported that LBW and delayed PNC are common to the Hispanic woman. In California, this cultural group comprises approximately 26% of the population, and much debate concerning health care reform has been targeted at this problem. A comparative study was conducted in California to examine the number of prenatal visits and the outcomes of Mexico-born Hispanics and U.S.-born Hispanics. Obstetric and medical record review for 783 women was done. The results show that more prenatal visits did not improve the outcome during pregnancy, labor, or the postpartum period. Because a large portion of PNC is now delivered by the advanced practice nurse, implications for practice include exploring alternatives for the delivery of culturally relevant care, addressing the barriers to caring for this population, and finding alternative models of care that have the potential to produce positive outcomes. PMID- 9203844 TI - Factors influencing the in vivo pharmacokinetics of peptides and antibody fragments: the pharmacokinetics of two PET-labeled low molecular weight proteins. AB - Monoclonal antibodies (MoAbs) were proposed as candidates for selective tumor targeting based on their high binding affinity for tumors and the absence of binding by normal tissue. However, the exclusive and complete transport of the drug have been found lacking in the use of intact MoAbs, especially in the case of solid tumors. Smaller fragments that maintained the desiderable tumor targeting characteristics appear to have an advantage because of the increase in whole body clearance and the shorter time to maximum target to non-target ratio. But the binding to normal tissue increases as the molecular weight or size decreases, especially in the kidney, because the glomerular sieving coefficient increases. We have used two approaches to overcome the normal tissue binding: a) the use of lysine to block the uptake of the Low Molecular Weight Proteins (LMWP) in the proximal tubules and b) the labeling of different amino acids in the LMWP to alter the residence time in the kidney. The combination of these two methods, blocking the uptake with lysine and shortening the residence time of the radioactive compound produced in the kidney, can lead to substantially decreased normal kidney targeting. The lysine paradigm was effectively demonstrated using 18F-labeled anti-Tac dsFv. Shortening the residence time was illustred by comparing the kinetics of the lysine catabolite versus the methionine metabolite. Furthermore, the rapid pharmacokinetics of the LMWP are consistent with the use of the shorter half-lives of PET radionuclides and the concomitant increase in quantitation and sensitivity afforded by PET. PMID- 9203845 TI - Regulatory peptide receptors as molecular targets for cancer diagnosis and therapy. AB - Regulatory peptides are small, readily diffusable and potent natural substances with a wide spectrum of receptor-mediated actions in humans. High affinity receptors for regulatory peptides such as somatostatin, substance P, vasoactive intestinal peptides, and cholecystokinin can be overexpressed in several human diseases, in particular in neoplasms, and represent therefore new molecular targets for cancer diagnosis and therapy. The availability of suitable regulatory peptide radioligands, which can be labeled with radioactive iodine or indium, makes peptide receptor scintigraphy a particularly useful new in vivo diagnostic tool, as seen with the example of somatostatin receptor scintigraphy (Octreoscan). The present article reviews the current in vitro data on regulatory peptide receptor expression in normal and diseased tissues, which represent the basis for the in vivo application of these molecules in nuclear medicine. PMID- 9203846 TI - Growth factor receptors as molecular targets for cancer diagnosis and therapy. AB - Growth factor receptors are of great interest as molecular targets for the diagnosis and treatment of cancer. Growth factor receptors are frequently over expressed on malignant cell populations since many cellular oncogenes encode either growth factors or their receptors. The wild-type epidermal growth factor receptor has a molecular weight of 170 kD and is over expressed on gliomas, bladder tumors, squamous cells carcinomas and breast carcinomas. Another growth factor oncogene, c-erbB-2, encodes a 185-kD glycoprotein found on the surface of gliomas, breast and ovarian cancers as well as other carcinomas of epithelial origin. In addition to causing over expression, oncogenic transformation also can result in genomic re-arrangements. An important example from the perspective of targeting is EGFRvIII, a deletion mutant which lacks amino acids 6-273 in the extracellular domain of the epidermal growth factor receptor. The EGFRvIII molecule (145 kD) may be of great value for targeting because it appears to be tumor-specific. Antibodies have been developed with specific reactivity with these growth factor receptors. Since these antibodies are internalized into the cell after receptor binding, it is necessary to use radiolabeling methods which residualize the radioactivity in the tumor cell after intracellular catabolism. To investigate this problem, we have evaluated the effect of radioiodination method on the in vitro and in vivo properties of an anti-EGFRvIII antibody. Methods studied were Iodogen, tyramine-cellobiose, and N-succinimidyl 5-iodo-3 pyridine-carboxylate with the last offering optimal localization in a human xenograft model. PMID- 9203847 TI - Radiopharmaceuticals to monitor gene transfer. AB - Advances in genetic engineering and molecular biology have opened the door to disease treatment by transferring genes to cells that are responsible for the pathological condition being addressed. These genes can serve to supplement or introduce the function of indigenous genes that are either inadequately expressed or that are congenitally absent in the patient. Additionally, however, they can introduce new functions such as drug sensitization, to provide a unique therapeutic target. Gene transfer is readily monitored in vitro using a range of histochemical and biochemical tests that are 'built in' to the therapeutic gene cassette. In vivo, in situ monitoring of the gene transfer and gene expression processes can be achieved with these tests only if biopsy is possible. Scintigraphic imaging can offer unique information on both the extent and location of gene expression, provided that an appropriate reporter gene is included in the therapeutic cassette. This overview includes a brief orientation to gene transfer therapy and is followed by a review of current approaches to gene therapy imaging. The concluding section deals with imaging based on radiolabelled nucleoside substrates for herpes simplex type-1 thymidine kinase, with emphasis on IVFRU, a stable, potent and selective HSV-1 TK substrate developed in our laboratories. PMID- 9203848 TI - Pharmacokinetic considerations in the development of oligomers as radiopharmaceuticals. AB - Single-stranded oligomers are attractive candidates for the next generation of radiopharmaceuticals because of their ability to bind specifically to their complementary single-stranded oligomers by hybridization. However, native, phosphodiester DNAs have been universally judged to be unsuitable because of excessive in vivo nuclease hydrolysis. Chemical modifications to phosphodiester DNAs are therefore required to improve pharmacokinetic properties before the potential of oligomers for radiopharmaceutical use can be realized. Fortunately, hundreds of modified oligomers have been prepared and tested, mostly in vitro, in connection with antisense chemotherapy. This chapter provides an overview of those results which are relevant to the use of the more important of these modified oligomers as radiopharmaceuticals. In brief, the phosphorothioate DNAs are stable in vivo but may be unsuitable in all forms because of high protein binding affinities which delay clearance and increase background radioactivity levels. The methylphosphonate DNAs are also stable but do not show high protein binding affinities. Like the vast majority of modified oligomers, they have not as yet been investigated as radiopharmaceuticals. However, it may be the synthetic oligomers which are the most attractive at present. In particular, PNA has been radiolabeled with 99mTc and shown in mouse studies to be stable, to clear rapidly without excessive protein binding and to hybridize to its complement in vivo. In conclusion, several oligomers display pharmacokinetic properties in preliminary studies which suggest that they deserve further consideration for use as radiopharmaceuticals. PMID- 9203849 TI - Pharmacokinetic modeling of multidrug resistance P-glycoprotein transport of gamma-emitting substrates. AB - P-glycoprotein, the human multidrug resistance (MDR1) gene product, is an integral membrane protein expressed on the plasma membrane of MDR tumor cells and is the best characterized of a family of efflux transporters that confer chemotherapeutic resistance. The use of gamma-emitting 99mTc-agents to image P glycoprotein function in human tumors in vivo has been proposed. Net tumor cell content of 99mTc-Sestamibi, 99mTc-Tetrofosmin and several 99mTc-Q-complexes 99mTc Q58 and 99mTc-Q63) are a function of passive potential-dependent influx and MDR1 P-glycoprotein-mediated active extrusion. To better understand the overall fidelity of these P-glycoprotein substrates to report MDR activity in vivo in relation to tissue perfusion, a compartmental model of tracer pharmacokinetics was developed. Modeling indicates that tissue perfusion will impact pharmacokinetics in vivo in a manner that will tend to diminish P-glycoprotein mediated phenotypic differences between tissues when they are perfusion-limited. However, dynamic imaging to extract efflux rate constants is independent of perfusion and may represent the highest quality methodology for collecting the desired information regarding activity of the efflux transposter. Much work remains to translate these concepts and biological targeting properties into clinical practice. PMID- 9203850 TI - Pharmacokinetic considerations in the development of peptide-based imaging agents. AB - Pharmacokinetic factors to be considered in the development of peptide-based imaging agents are reviewed. These include size, plasma protein binding, lipophilicity, resistance or susceptibility to proteolysis and radiolabel integrity. These factors are discussed in the context of several examples of thrombus and tumor imaging agents either commercially available or in development. PMID- 9203851 TI - Pharmacokinetic considerations in the PET and SPECT evaluation of CNS receptors. AB - Positron emission tomography (PET) and single-photon emission computed tomography (SPECT) are the only functional imaging methodologies that allow to evaluate, in vivo in human, specific binding proteins such as receptors, transporters or enzymes. PET and SPECT have already proved to be unique tools to follow, in the living human brain, the kinetics of the interaction of a radiolabelled ligand with its receptors. However, these imaging techniques measure the radioligand concentration in regions of interest (ROIs) as a function of time but they do not allow the direct measurement of the binding parameters, i.e. receptor concentration and radioligand affinity. To estimate these physiological parameters a mathematical model must be designed to simulate the kinetics of the radioligand. The modelling of the data obtained using such equilibrium or dynamic models allow to extract from the kinetic data these physiological parameters. PET and SPECT imaging methodologies have then opened a new era in brain biochemistry and have already important applicants in brain physiopathology, clinical pharmacology and drug development. PMID- 9203852 TI - Prodrugs in site-selective delivery of radiopharmaceuticals. AB - This paper reviews basic rules for the design of site-selective prodrugs and various modes of their activation with particular emphasis on the applications of prodrugs to targeted delivery of radiopharmaceuticals. Although many radiopharmaceuticals are "targeted" to specific tissues or organs, we will discuss only agents that are either chemically or metabolically transformed producing an active form that is retained by its target. Site-specific prodrugs of diagnostic radiopharmaceuticals are routine in the nuclear medicine applications, but the instances of targeting of radiotherapeutic prodrugs are surprisingly rare. We have concentrated on our own efforts to design and synthesize site-selective prodrugs of 5-[125I]iodo-2'-deoxyuridine (125IUdR) for cancer radiotherapy. The prodrugs of 125IUdR for targeted delivery include several derivatives with altered permeability, 3',5'-dioctanoyl, 3',5'-dioleoyl, 3'- and 5'-N-alkyl-dihydropyridyl, 3'- and 5'-N-alkyl-dihydroisoquinolyl, and 3'- and 5'-N-alkyl-dihydroacridinyl esters of 125IUdR; polymeric and macromolecular prodrugs of 125IUdR for a carrier-mediated or local delivery; metabolically trapped 125IUdR prodrugs; and glycoconjugate prodrugs for oral colon-specific delivery of 125IUDR, 125IUDR-5'-beta-d-cellobioside, 125IUDR-5'-beta-D glucopyranoside, 125IUDR-5'-beta-D-galactopyranoside and 125IUDR-5'-beta-D glucuronide. We also describe prodrugs of several diagnostic agents in the context of the metabolic trapping as the primary targeting modality. For various diagnostic agents the prodrug target-associated enzymes are discussed and examples of the site-specific release of the active agent are given. A brief overview of an emerging role of residualizing labels in radioimmunotherapy is included. PMID- 9203853 TI - Pharmacokinetic considerations in the development of labeled liposomes and micelles for diagnostic imaging. AB - The current status of application of liposomes and micelles as carriers for diagnostic imaging agents in experimental and clinical medicine is considered. Liposomes and micelles loaded with appropriate contrast agents have been shown to be suitable for all imaging modalities, including gamma-, magnetic resonance (MR), and computed tomography (CT). The methods are briefly described to prepare liposomes loaded with various contrast agents, as well as some basic data on their in vitro and in vivo properties and biodistribution. Certain pharmacokinetic considerations associated with the use of plain and long circulating liposomes and micelles as pharmaceutical carriers are discussed. The application of contrast-loaded liposomes in different modalities for the experimental and clinical imaging of various organs, tissues, and pathological conditions is briefly reviewed. New trends in the preparation and use of contrast loaded liposomes and micelles are also considered, such as the application of amphiphilic polychelating polymers and polymers for steric protection of microparticulate pharmaceutical carriers. PMID- 9203854 TI - Can receptors be imaged with MRI agents? AB - A review of the feasibility of imaging receptors was made using published literature. The results suggest that there is no physical limitation to imaging the classical biochemical receptors using currently available compounds. Limitations occur because of biological constraints. These constraints are those of delivery of contrast material to the site of the receptor in sufficient quantities and the biological implications of saturating receptors. These limitations are reduced by improving the physics through increasing the relaxivity of the contrast agent either by greater intrinsic relaxivity or by attaching many relaxing agents to the ligand. Biochemical constraints are reduced by targeting receptors involved with transport systems such as receptor mediated endocytosis and by targeting those sites that are in or readily accessible to the vascular system. Once targeted MRI agents are developed their clinical use will, most probably, be different than that of the corresponding radiopharmaceuticals. PMID- 9203855 TI - The role of radiopharmacological imaging in streamlining the drug development process. AB - Radioimaging techniques have found a place in clinical diagnosis, but there has been a hesitancy to use this approach in drug development. This reluctance may have been due to the availability of ligands, the time and cost of synthesis and the number of centres and, although these perceived problems have been largely overcome, for many the benefits are not evident. The use in drug development is potentially large since tomography can measure drug levels, specific binding, blood flow and activity within the human body. In drug discovery, combinational chemistry and high throughput screening, the synthesis of candidate drugs with specific binding properties are dependent on understanding the disease and using appropriate in vitro or animal models. Using small animal tomographs, these can be validated using radioimaging. Pharmacokinetics and metabolic problems, such as the distribution of inhaled gases, drug targeting into tumours of the brain or specific gastrointestinal absorption sites can be investigated within the human rather than relying on animals. The high specific activity allows low doses to be administered to man with limited safety studies, permitting kinetic and metabolic studies to be undertaken early in development. Safety studies and ensuing toxicological endpoints in animals rely on histopathology for gross degenerative in physiological function. Where concern exists, radioimaging could detect early in situ changes in humans, for example hepatic toxicity, before they become hazardous. In clinical studies, the action of drugs can be measured directly at the effector site prior to undertaking longer studies, which is important for many diseases, but particularly for those such as Alzheimer's disease, where improvements may be slow or subtle. PMID- 9203856 TI - Positron emission tomography in drug development. AB - There are four kinds of measurements that can be carried out with positron emission tomography (PET) that can contribute significantly to the process of drug development: pharmacodynamic measurements of tissue metabolism influenced by a given drug; precise measurements of tissue blood flow; tissue pharmacokinetics of a given drug following administration of a particular dose; and the temporal course of ligand-receptor interaction. One or more of these measurements can greatly improve the decision making involved in determining the appropriate dose of a drug, the clinical situations in which a drug might be useful, and the linkage of pharmacokinetics with pharmacodynamics, which is at the heart of effective drug development. The greater the potential of a particular compound as a therapeutic agent, the greater the potential for PET to contribute to the drug development process. PMID- 9203858 TI - Commodification, commercialization, and embodiment. PMID- 9203857 TI - Assisted reproductive technologies, ads, and ethics: philosophical, ethical, and clinical perspectives on the use of advertising in reproductive medicine. A conference held by the National Advisory Board on Ethics in Reproduction. PMID- 9203859 TI - Ethics, advertising, and assisted reproduction: the goals and methods of advertising. PMID- 9203860 TI - The commodification and advertising of infertility treatment. PMID- 9203861 TI - Accountability, representation, and advertising. PMID- 9203862 TI - Innovations in infertility treatment and the rush to market. PMID- 9203864 TI - Panel one: marketing strategies and informing the patient/consumer. A fertility center describes its shared-risk program. PMID- 9203863 TI - Accountability in the advertising and marketing of assisted reproduction. PMID- 9203865 TI - Panel One: marketing strategies and informing the patient/consumer. Infertility diagnostic techniques: the rush to market. AB - The human genome project will result in many new diagnostic techniques applicable to assisted reproduction. These may be directed both at infertile patients as well as at patients who seek ART because they carry a deleterious genetic disease. There are many potential advantages to the use of preprocedural testing as a means of obtaining additional preliminary information that might allow patients to make a more informed choice, either when deciding between ART and adoption or when comparing the benefits of various ART procedures. The cost of testing is relatively low when compared with the cost of additional IVF or ICSI cycles. In the case of genetic screening protocols, the results may be useful not only to the patient but also to the next generation. Genetic counseling should be made available in concert with expanded opportunities, both for diagnosing infertility and for making preimplantation genetic diagnoses. Already some of the genetic screening that has occurred because of discoveries made during the human genome initiative has become a regulatory concern with regard to insurance availability and other issues. When counseling patients, it is important to point out that certain tests hold far more prognostic value and/or serious health implications than others. A test to diagnose breast cancer susceptibility, for example, holds more serious ramifications than the less predictive test that detects the phenotype for Klinefelter's syndrome. Presymptomatic prognostic screening systems, such as the tests currently used to detect genetic mutations related to breast and prostate cancer and cystic fibrosis, provide diagnostic clues but also have serious societal implications. We are on a rather slippery slope when we attempt to determine which tests actually might prove beneficial to patients, both individually and collectively. Therefore, as part of patient counseling, cost-benefit ratios, in addition to clinical and preventive implications, should be weighed. PMID- 9203866 TI - Panel one: marketing strategies and informing the patient/consumer. Infertility, advertising, and choice: a consumer advocate's perspective. PMID- 9203867 TI - Panel two: reporting and advertising success rates--the Gordian Knot of assisted reproductive technology. The role of professional societies. PMID- 9203868 TI - Panel two: reporting and advertising success rates--the Gordian Knot of assisted reproductive technology. The role of a federal regulatory agency. PMID- 9203869 TI - Age-related changes in human anterior cruciate ligament (ACL) collagen fibrils. AB - The correlation between anterior cruciate ligament (ACL) collagen fibril diameter and aging were studied in subjects aged 15 to 87 years. The samples processed for light and electron microscopy showed statistically significant differences in collagen fibril diameter among young (< 20 years), adult (20-60) and elderly subjects (> 60 years). In the young, the ACL was made up of collagen fibrils which were highly variable in size (range 20-180 nm); the diameter distribution curve was very asymmetrical (mean asymmetry +0.895). In adults and elderly subjects, the maximum diameter had decreased remarkably (120 and 110 nm, respectively) and the diameter distribution curve had become less asymmetrical (mean asymmetry +0.527 and +0.297 respectively). Fibril concentration increased considerably from young (68 fib/mu 2) and elderly subjects (140 fib/mu 2). This reduction in diameter and the relative change in collagen fibril concentration may be related to changes in elastic stiffness. The increase in small collagen fibrils and the marked rise in their concentration may make the ligament more pliable. These findings are similar to those we obtained in Achilles tendon. They demonstrate that both ACL and Achilles tendon, a tissue which responds to unidirectional mechanical forces more than ACL does, show a reduction in diameter value during ageing. These data further suggest that collagen fibril diameter is related to the aging process. PMID- 9203870 TI - Collateral circulation in distal occlusion of lower limb arteries: an anatomical study and statistical research in 40 elderly subjects by echo-color-Doppler method. AB - The collateral circles and their hemodynamic significance in distal lower limb arterial occlusion have been described in an elderly population. Overall 40 subjects (20 men and 20 women; age range 66-83) with symptomatic lower limb arteriopathy (Fontaine's stage II) have been studied combing Contrast Angiography and Color Doppler Echography of the lower limb arterial district. In our population, the results showed that the tibialis arteries were the vessels most often involved in arterial occlusion (posterior tibialis a., 15 cases = 37.5%, posterior tibialis a., 12 cases = 30%), followed by the peroneal a. (8 cases = 20%) and by the popliteal a. (5 cases, 12.5%). In the occlusion of the popliteal artery the collateral circle was mainly established through the deep femoral a., the great anastomotic a., the recurrent posterior tibialis a., and from the articular supero-lateral a. In the occlusion of the anterior tibialis artery the collateral circulation was ensured through the collateral posterior tibialis as. and through the collateral perineal as. In the occlusion of the posterior tibialis a., the collateral circle was established through the great anastomotic a., through the branchers of the arterial circle of the ankle and from the perforating plantar as. (anterior tibial a.). Finally, in the occlusion of the peroneal a., the collateral circulation was only represented by branches of the arterial circle of the ankle. The hemodynamic compromission, measured by the Windsor Index, was the highest for popliteal occlusions (mean IW = 34.3%). Occlusions of the anterior tibialis a. (mean IW. = 35.48%), of the peroneal a. (mean IW = 44.71%), and of the posterior tibialis a. (mean IW = 55.44%) showed progressively lower hemodynamic compromission. Gender differences in hemodynamic significance at each level of occlusion were not significant. PMID- 9203871 TI - The structure of bone tissues and the cellular control of their deposition. PMID- 9203872 TI - The morphofunctional organization of lymphatic vascular periphery. PMID- 9203873 TI - [The Bengmark principle. Placement and fixation of jejunal catheters in enteral feeding]. PMID- 9203874 TI - Paediatric anaesthesia--who should do it? The view from the specialist hospital. PMID- 9203875 TI - Paediatric anaesthesia--who should do it? The view from the district general hospital. PMID- 9203876 TI - Neonatal welfare and placental transfer of fentanyl and bupivacaine during ambulatory combined spinal epidural analgesia for labour. AB - To investigate current concerns that potent opioid drugs, such as fentanyl, used for labour regional analgesia may affect neonatal status, maternal and umbilical plasma concentrations of fentanyl and bupivacaine at delivery were measured in 40 nulliparous patients receiving low-dose combined spinal epidural analgesia. Neonatal assessments included Apgar scores, umbilical blood gases and neurobehavioural tests. All maternal and umbilical venous plasma concentrations were low. Maternal and umbilical vein total fentanyl concentrations increased with increasing doses of epidural fentanyl (r = 0.46 and 0.30, respectively, p < 0.01). There were no significant differences between maternal and umbilical venous plasma total or free concentrations of fentanyl. Mean umbilical vein/maternal fentanyl ratios were 1.12 for total drug and 1.20 for free drug and values were unrelated to the last epidural bolus to delivery interval (r = 0.12, p = 0.49). There were no correlations between Apgar scores, umbilical blood gases or neurobehavioural scores and umbilical venous concentrations of either fentanyl or bupivacaine. The dose of fentanyl used for ambulatory combined spinal epidural analgesia would appear to have a negligible effect on neonatal condition. PMID- 9203877 TI - Patient-maintained propofol sedation. Assessment of a target-controlled infusion system. AB - We have developed a system which allows patients to operate a target-controlled infusion of propofol to provide sedation and we have studied its use in 36 unpremedicated patients undergoing local and regional anaesthetic procedures lasting 10-280 min. An intravenous propofol infusion was started at a target plasma level of 1 microgram.ml-1. The patient was able to increase the target propofol concentration in 0.2- microgram.ml-1 increments by pressing a demand button. There was a lockout interval of 2 min and a maximum permissible target concentration of 3 micrograms.ml-1. There was considerable interindividual variability in propofol consumption (mean 39.3 micrograms.kg-1.min-1, range 3-131 micrograms.kg-1.min-1), no cardiovascular instability and little oversedation. Eight patients required supplementary oxygen. Optimal sedation was provided at median target concentrations of 0.8-0.9 microgram.ml-1. The target-controlled infusion system bias was-47% and the inaccuracy was 48%. Patient satisfaction was high and 89% said that they would definitely use the technique again. This technique combines the benefits of target-controlled infusion with patient controlled feedback and produces safe intra-operative sedation. PMID- 9203878 TI - Heterogeneity in intensive care units: fact or fiction? AB - Reports and guidelines concerning intensive care practice have been issued recently. However, the introduction of such centrally issued recommendations may be difficult because of marked heterogeneity between intensive care units. This study examined the facilities (number of beds, consultant sessions, nursing establishment), annual workload (number and types of patients admitted) and outcome (intensive care unit mortality) in the (old) Anglia Region. There were significant differences in the distribution of patients' ages, severities of illness, diagnoses, durations of admission and outcomes. Such heterogeneity may make multicentre trials more difficult to conduct and create problems when uniform measures designed to improve intensive care services are being planned. PMID- 9203879 TI - Factors affecting neostigmine reversal of vecuronium block during sevoflurane anaesthesia. AB - We examined the influence of the concentration of sevoflurane and the degree of muscle block at the time of reversal on the activity of neostigmine. Ninety ASA 1 2 patients were anaesthetised with 0.2, 0.7 or 1.2 MAC of sevoflurane (30 patients each) in 66% nitrous oxide in oxygen. The electromyographic (EMG) response of the adductor digiti minimi was monitored at 20-s intervals after train-of-four stimulation of the ulnar nerve. The initial neuromuscular block was produced by vecuronium 100 micrograms.kg-1. When the amplitude of the first response (T1) values had recovered to 10%, 25% or 40% of the control, neostigmine 40 micrograms.kg-1 was administered. The train-of-four ratio values were recorded at 1-min intervals during the subsequent 15-min period. Higher endtidal concentrations (p < 0.0001) and more pronounced block at the time of reversal (p < 0.0001) were associated with a delayed recovery in the train-of-four ratio. In addition, the train-of-four ratio 15 min after neostigmine administration was more dependent on the sevoflurane concentration than on the degree of block present (p < 0.0001). These results confirm that neostigmine (40 micrograms.kg-1) can reverse vecuronium-induced but not sevoflurane-induced neuromuscular block. PMID- 9203880 TI - Propofol anaesthesia and vomiting after myringoplasty in children. AB - To determine whether propofol anaesthesia reduces the incidence of nausea and vomiting after ear surgery, 40 children aged 4-16 years were randomly assigned to receive either propofol or inhalational anaesthesia. Children in the propofol group had anaesthesia induced with propofol and maintained with propofol-nitrous oxide and those in the inhalational group had anaesthesia induced with thiopentone and maintained with isoflurane-nitrous oxide. Nausea and vomiting were recorded for 24 h after surgery and metoclopramide was offered to children who vomited more than twice. We found that 11 children (55%) who had propofol and 14 children (70%) who had inhalational anaesthesia vomited one or more times after surgery (difference not significant). The incidence of vomiting was lower in the propofol group during the first 2 h after surgery (0% and 25% propofol and inhalational groups, respectively) (p < 0.05) but was similar at all other time intervals. Rescue anti-emetic was given to two (10%) and eight (40%) children in the propofol and inhalational groups, respectively (p < 0.05). We conclude that propofol anaesthesia alone is not an effective means of preventing vomiting after middle ear surgery in children. PMID- 9203882 TI - The use of paraglossal straight blade laryngoscopy in difficult tracheal intubation. AB - In 10 cases of unexpected difficult tracheal intubation using the Macintosh laryngoscope blade, the larynx could not be seen. In each case, a good view was achieved using the Miller blade lateral to the tongue, which enabled tracheal intubation under direct vision. The results achieved using narrow, low-profile straight laryngoscope blades with this technique are reviewed. The improved view obtained with this technique is a consequence of reduced tongue compression as compared with the Macintosh technique. This leads both to an improved line of sight, and to a reduced risk of backward displacement of the tongue and epiglottis. In addition, the molar or retromolar variation of the technique reduces the intrusion of maxillary structures into the line of sight, so that a better view of the larynx is achieved for a given degree of soft tissue compression. Paraglossal straight blade laryngoscopy may have an advantage over use of the Macintosh technique when intubation proves unexpectedly difficult. It is perhaps time to question standard teaching about the role of the curved blade in such patients or, more particularly, whether the technique of laryngoscopy as currently taught is optimal. The paraglossal straight blade technique needs to be practised in routine intubation before it can be used with confidence in difficult cases. PMID- 9203881 TI - Postoperative analgesic effect of intrathecal neostigmine and its influence on spinal anaesthesia. AB - A clinical trial was conducted to evaluate the postoperative analgesic efficacy and the safety of intrathecal neostigmine in patients undergoing anterior and posterior vaginoplasty under spinal anaesthesia. Thirty-six patients were randomly divided into three groups to receive: normal saline (1 ml), morphine (100 micrograms in 1 ml of saline) or neostigmine (100 micrograms in 1 ml of saline) intrathecally just before a spinal injection of hyperbaric bupivacaine (0.5%, 4 ml). The mean [SD] time to the first analgesic (nonsteroidal anti inflammatory drug) administration was significantly prolonged by intrathecal neostigmine (10.7 [4.3] h) and morphine (15.3 [3.0] h) compared with saline (4.5 [1.0] h). The three groups also differed in the number of patients requiring subcutaneous morphine to complement the analgesia provided by the intramuscular nonsteroidal anti-inflammatory drugs and the mean [SD] times for their administration: eight patients in the saline group (8.0 [3.8] h), one patient in the morphine group (18 h) and two patients in the neostigmine group (8 and 12.9 h). The morphine and neostigmine groups showed similar analgesic effectiveness. The characteristics of spinal anaesthesia were not modified by intrathecal morphine or neostigmine. Severe nausea and vomiting, sweating and distress during surgery were the most obvious adverse effects of intrathecal neostigmine. On the other hand, less hypotension was observed in the neostigmine group. The usefulness of intrathecal neostigmine as the sole postoperative analgesic may be restricted by the severity of its adverse effects. PMID- 9203883 TI - Nitric oxide: description of a pipeline delivery system. AB - We describe a pipeline system suitable for the delivery of nitric oxide gas to an 18-bed intensive care unit. The pipeline was developed and installed according to the current UK regulations HTM 2022, which relates to the supply of piped medical gases. Where HTM 2022 did not specify the appropriate standard, we consulted widely to achieve a safe solution. We continue to monitor all aspects of the performance of the pipeline to ensure safe operating practices and recommend changes to the standards. PMID- 9203884 TI - Leakage of fluid around low-pressure tracheal tube cuffs. AB - The aim of the study was to evaluate leakage of liquid past the low-pressure cuffs of tracheal tubes. Ten samples of each of nine different types of tubes were tested in a PVC mock trachea, using intracuff pressures of 20, 30, 40 and 50 cmH2O. In five types of tubes, 6-10 cuffs allowed profuse leakage (> 20 ml water in 5 min) even at the highest intracuff pressure, i.e. 50 cmH2O. In the most efficient tube, all the cuffs were leak-proof (leakage < 5 ml in 5 min) at 40 cmH2O and in the second best type the cuffs were leak-proof at 50 cmH2O. The leakage of fluid past the tracheal tube remains an unresolved problem with low pressure cuffs. PMID- 9203885 TI - Latex allergy: an emerging clinical and occupational health problem. AB - Allergy to latex products has become a problem of increasing dimensions, affecting both patients and health-care professionals. Certain high-risk groups and sources of latex exposure have been identified. The clinical manifestations vary from contact eczema through a range of allergic syndromes including life threatening anaphylaxis. An understanding of the clinical and immunological features of latex allergy provides the basis of therapeutic and preventive strategies. Latex allergy is also becoming a major occupational health issue. PMID- 9203886 TI - Epidural abscess following epidural steroid and local anaesthetic injection. AB - Epidural abscess is a well-recognised but rare complication of epidural catheter placement. We have found only five previous reports of epidural abscess from noncatheter-related administration of steroids and/or local anaesthetic. We describe a further case which led to critical illness and emphasise the association between diabetes mellitus and epidural infection. PMID- 9203887 TI - Caesarean section in a parturient with respiratory failure caused by cystic fibrosis. AB - We describe a 27-year-old primigravida suffering from cystic fibrosis. Her chest was colonised with Burkholderia cepacia and she was in respiratory failure for which she required constant nasal intermittent positive pressure ventilation. In view of her rapid deterioration, Caesarean section was performed under epidural anaesthesia at 25 weeks gestation. A live 790-g boy was delivered. Post operatively she made steady progress for 5 days although still requiring nasal ventilatory support. Thereafter she developed pneumonia and required tracheal intubation and ventilation on the eighth day. Her increasing hypoxaemia and pulmonary hypertension failed to respond to any therapy including inhaled nitric oxide and she died on the tenth postoperative day. PMID- 9203888 TI - Complete airway obstruction during awake fibreoptic intubation. AB - Awake fibreoptic intubation is well established as the optimum method of securing the airway in patients in whom difficulty is anticipated. We report a patient undergoing awake fibreoptic intubation in whom the use of topical local anaesthetic precipitated acute loss of the airway so that urgent surgical intervention was required. PMID- 9203889 TI - The effect of single-handed cricoid pressure on neck movement after applying manual in-line stabilisation. AB - In 30 ASA 1 and 2 patients undergoing general anaesthesia and neuromuscular paralysis, manual in-line stabilisation of the neck in a neutral position was performed and single-handed cricoid pressure was applied. Vertical displacement was measured from the midpoint of the neck (directly below the cricoid cartilage). Measurements were also made at the tragus of the ear and the shoulder; both of which acted as fixed reference points. Mean neck displacement was 4.6 mm with a range of 0-8 mm. Mean tragus and shoulder displacements were 0.5 mm and 0.9mm, respectively, with a range of 0-2 mm at each point. Vertical displacement was also measured in 10 patients from a stylet fixed to the posterior aspect of the neck. Mean displacement measured at this point was 5.0 mm with a range of 2-9 mm. Single-handed cricoid pressure caused vertical displacement of the neck of between 4.6 and 5 mm with a range of 0-9 mm. Only some of this movement, i.e. 0.5-0.9 mm (range 0-2mm) can be accounted for by displacement of the whole patient as determined from measurements at the two fixed reference points. These findings have implications for emergency management of the airway in trauma patients. PMID- 9203891 TI - Laryngeal mask lubrication. A comparative study of saline versus 2% lignocaine gel with cuff pressure control. AB - We compared 2% lignocaine gel with saline as a lubricant for the laryngeal mask airway in 126 patients receiving positive pressure ventilation in whom cuff pressures were limited to 60 cmH2O and peak airway pressures to less than 17 cmH2O. The incidence of sore throat was similar for both groups and there were no emergence problems. There were significantly more intra- and postoperative complications in the lignocaine group (p < 0.05) but the frequency of sore throat was similar when the device was inserted at the first attempt. Positive pressure ventilation to normal end-tidal CO2 values was possible in all patients. Lignocaine gel is an unsuitable lubricant for the laryngeal mask airway. Cuff pressure limitation to 60 cmH2O does not necessarily impede ventilation and may be an important factor in reducing emergence and postoperative complications. PMID- 9203890 TI - Body position and late postoperative nocturnal hypoxaemia. AB - Thirteen patients were monitored for nocturnal body position (supine vs. side) and arterial oxygen saturation pre-operatively and on the second postoperative night after major abdominal surgery. The number of positional changes were significantly decreased after operation (p < 0.05) with a trend towards more time spent in the supine position (p = 0.1). Individual mean arterial oxygen saturation decreased postoperatively (p < 0.05) but without a difference between the supine and side positions (p = 0.9). Pre-operatively, episodic desaturations were significantly more frequent in the supine position than on the side (p < 0.05) but not postoperatively. Pain was the most frequent reason for decreased nocturnal movements. PMID- 9203892 TI - The effect of glossopharyngeal nerve block on pain after elective adult tonsillectomy and uvulopalatoplasty. AB - This controlled, randomised, double-blind study compared whether glossopharyngeal nerve block and intravenous morphine administered peri-operatively, decreased pain following elective adult tonsillectomy and uvulopalatoplasty more than morphine alone. Sixteen of 30 patients undergoing uvulopalatoplasty and 38 of 78 patients having tonsillectomy received bilateral glossopharyngeal nerve blocks, using bupivacaine 0.5% and adrenaline 1:200,000, or no intervention. There were no differences in postoperative analgesic consumption between the two groups. Visual analogue pain scores measured during swallowing in the recovery room and on the ward postoperatively were significantly less overall in uvulopalatoplasty patients who had received a block (p = 0.004). This difference was not found for tonsillectomy. We found no significant differences between groups, in pain scores recorded during the first 5 days at home. We conclude that glossopharyngeal block does not improve analgesia following tonsillectomy although there is short-lived benefit following uvulopalatoplasty. PMID- 9203893 TI - Adverse response to sevoflurane induction followed by enflurane maintenance. PMID- 9203894 TI - Pain following craniotomy. PMID- 9203895 TI - Does at risk mean acceptable? PMID- 9203896 TI - Sciatic nerve injury and caesarean section. PMID- 9203897 TI - Remifentanil for major abdominal surgery. PMID- 9203899 TI - Introducer for the reinforced laryngeal mask airway. PMID- 9203898 TI - End-tidal oxygraphy and safe duration of apnoea in young adults and elderly patients. PMID- 9203901 TI - Brachial artery catheterisation in paediatric intensive care. PMID- 9203900 TI - Use of auditory feedback to aid identification of the epidural space. PMID- 9203903 TI - Dantrolene infusion in severe tetanus. PMID- 9203902 TI - An unpredictable and possibly dangerous hazard of an anaesthetic scavenging system. PMID- 9203904 TI - Digital assistance for the anaesthetist. PMID- 9203905 TI - Modification to the ventilation-exchange bougie. PMID- 9203906 TI - Antiphospholipid syndrome and neurosurgery. PMID- 9203907 TI - Plasmapheresis in neuroleptic malignant syndrome. PMID- 9203908 TI - Nalbuphine and pruritus. PMID- 9203909 TI - Intrapulmonary shunt during one-lung anaesthesia. PMID- 9203910 TI - Superior and inferior vena cava obstruction in leucocytoclastic vasculitis. PMID- 9203911 TI - Spontaneous movement after injection of propofol. PMID- 9203912 TI - Abuse of 'sharps boxes'. PMID- 9203913 TI - Anaesthesia, anaesthetics and anaesthesiology. PMID- 9203914 TI - The European Board of Surgery Qualification (EBSQ) was born in 1996. AB - The EU has sent delegates to the Section of Surgery, UEMS and the EBS for some years. The EBSQ is the only way to harmonise surgical standards across the EU. It is recognised that standards very across Europe and although CCST must be recognised across Europe, in practice there are 15 CCSTs of different standards. UK and Ireland have surgical training programmes and standards which are admired and the EBSQ has been centred in London and the sterling area. There is therefore a unique opportunity to influence European surgery. What is far from certain is whether UK trainee surgeons will feel the need to take EBSQ, for them it is very much an optional extra. PMID- 9203915 TI - The demise of the DGH. Acute general hospital surgical services of the future. PMID- 9203916 TI - Out of hours cross-cover between oral and maxillofacial surgery and ear, nose and throat surgery. AB - Emergency cross-cover between two specialties, oral and maxillofacial surgery and ear, nose and throat surgery, has been introduced and run successfully in York District Hospital for the last 18 months in order to reduce the working hours of junior doctors, as recommended by the New Deal. During this trial period of out of normal hours cross-cover in a two-tier system, we have found that the new arrangement has increased our shared knowledge, understanding and practical experience of both specialties as well as satisfying the service needs with no significant clinical problems arising. PMID- 9203917 TI - The hidden cost of audit. AB - Although a considerable sum of money has been made available for the introduction of medical audit, many of the indirect costs have not been considered. We have assessed one of these areas, the medical audit meeting, by calculating the cost to Preston Acute Hospitals (PAH) Trust and extrapolating it to England and Wales. A potential cost of Pounds 678,000 per annum to PAH Trust could be reduced to 14 per cent of this figure if current practices were changed. The hidden cost for England and Wales could be up to Pounds 106 million per annum. The cost of medical audit meetings could be dramatically reduced if current practices were modified. This could be done with little detriment to the audit process. PMID- 9203918 TI - Potential availability of patients in a short stay ward for medical student teaching. AB - The trend towards shorter hospital in-patient stays has decreased the availability of patients for undergraduate medical student teaching. We surveyed 100 consecutive short stay surgical inpatients admitted to hospital on the day of planned operation to determine whether they might be available for student teaching before surgery. We found that there was a median delay of three hours (standard deviation +/-2.3 hours) between the completion of all medical and nursing procedures and the departure of the patient for the operating theatre. All patients were asked if they would agree in principle to participate in student teaching. 98 per cent said they would be willing to be taught upon. We argue that surgical patients attending the short stay ward are a valuable potential source of teaching material. PMID- 9203919 TI - Overseas trainees--training in the UK. A report on the First Surgical Conference for Overseas Trainees organised by the University of Liverpool and held at the Tropical School of Medicine, Liverpool in December, 1995. PMID- 9203920 TI - Surgical training of overseas qualified doctors. Are we training the wrong doctors at the wrong time in the wrong way? PMID- 9203922 TI - Surgery--the female deficit. PMID- 9203921 TI - The life and work of David Slome. PMID- 9203923 TI - The effect of the 'Calman' Report on academic aspects of surgical training. PMID- 9203924 TI - The origin of the word 'stent'. PMID- 9203926 TI - The surgeon's assistant: an orthopaedic model at Hastings. PMID- 9203927 TI - Use of a commercial computer program for continuous audit of surgical complications. PMID- 9203925 TI - Should consultants see new or follow-up outpatients? PMID- 9203928 TI - Reporting of clinical trials in Gut: the CONSORT statement. Consolidated Standards of Reporting Trials. PMID- 9203929 TI - Laparoscopic surgery for gastro-oesophageal reflux disease. PMID- 9203930 TI - Hepatitis B virus infection and liver transplantation. PMID- 9203931 TI - The genetics of inflammatory bowel disease. PMID- 9203932 TI - Cholecystokinin in transient lower oesophageal sphincter relaxation due to gastric distension in humans. AB - BACKGROUND AND AIMS: Transient lower oesophageal sphincter relaxations (TLOSRs) has been found to be the main mechanism of gastro-oesophageal reflux. In dogs, cholecystokinin (CCK) is involved in their occurrence. The aim was to evaluate the role of endogenous and exogenous CCK in the occurrence of TLOSRs induced by gastric distension at constant pressure in humans. METHODS: Ten healthy volunteers were studied. Lower oesophageal sphincter pressure was monitored with a sleeve device and gastric distension was performed via an intragastric bag monitored by a barostat. During distensions, saline, CCK (30 ng/kg/h) or the CCK A receptor antagonist loxiglumide (10 mg/kg/h) was perfused in a random double blind order. RESULTS: There was no significant difference between the number of TLOSRs during the different distensions with saline; CCK increased the number of TLOSRs at a mean rate of 13.1 compared with 9.1 with saline (p < 0.001). Loxiglumide significantly decreased the number of relaxations to 5.3 versus 8.3 under paired saline infusion (p < 0.001). CONCLUSIONS: In humans, CCK-A receptor subtype is involved in the occurrence of transient lower oesophageal sphincter relaxations induced by gastric distension. PMID- 9203933 TI - Reappraisal of bicarbonate secretion by the human oesophagus. AB - BACKGROUND AND AIMS: Administration of omeprazole to healthy volunteers was recently reported to increase proximal duodenal mucosal bicarbonate secretion. As human oesophagus also secretes bicarbonate, the hypothesis was tested that omeprazole may stimulate oesophageal bicarbonate secretion and thus contribute to the therapeutic efficacy of the drug in gastro-oesophageal reflux disease. SUBJECTS AND METHODS: In nine healthy volunteers, oesophageal "steady state" perfusion of a 10 cm open segment of distal oesophagus was performed twice in random order. The volunteers were pretreated with either 60 mg/day omeprazole for three days and 80 mg intravenous omeprazole before perfusion or 600 mg/day ranitidine for three days and 50 mg/h intravenously during the perfusion. Saliva and samples of aspirate from the perfused oesophagus and stomach were collected and bicarbonate concentrations were measured. RESULTS: The median rates (95% confidence intervals) of intrinsic oesophageal bicarbonate secretion, corrected for contaminating salivary and gastric bicarbonate, were 89 (33-150) and 121 (63 203) mumol/h/10 cm (p > 0.5) in omeprazole and ranitidine treated subjects respectively. Salivary and gastric bicarbonate contaminating the oesophagus accounted for 14% and 3%, respectively, of total oesophageal bicarbonate output. CONCLUSIONS: Bicarbonate secretory capacity of the human oesophagus is less than previously assumed, and the clinical relevance of intrinsic oesophageal bicarbonate for mucosal defence may be overestimated. As omeprazole and ranitidine did not affect bicarbonate secretion differently there was no evidence that omeprazole acts on bicarbonate secretory cells in the oesophageal mucosa. PMID- 9203934 TI - Double blind cross-over placebo controlled study of omeprazole in the treatment of patients with reflux symptoms and physiological levels of acid reflux--the "sensitive oesophagus". AB - BACKGROUND: At least 10-15% of patients with reflux symptoms have a normal endoscopy and physiological levels of acid reflux on pH monitoring. Such patients with 50% or more of symptoms associated with acid reflux episodes have "a positive symptom index" (SI), and it has been proposed that this defines the "sensitive oesophagus". AIM: To test the response to omeprazole 20 mg twice daily for four weeks of patients with normal levels of acid reflux using a randomised, placebo controlled, double blind, cross-over design. PATIENTS: Eighteen patients with normal levels of reflux, 12 of whom had a positive SI. METHODS: Response was measured by symptomatic assessment and the SF-36 quality of life (QOL) questionnaire. RESULTS: Patients with a positive SI showed the following improvements on omeprazole compared with placebo: decrease in symptom frequency (p < 0.01), severity (p < 0.01) and consumption of antacids (p < 0.01). In the group with a negative SI only one patient clearly improved. The QOL parameters for bodily pain (65.6 v 53.4, p = 0.03) and vitality (60.6 v 48.8, p = 0.049) were significantly better on omeprazole than placebo for the group overall. CONCLUSION: Omeprazole improves symptoms in 11 of 18 patients with normal endoscopy and pH monitoring, particularly those with a positive SI. This supports the theory that such patients have an oesophagus which is "sensitive" to acid reflux and are part of the GORD spectrum. PMID- 9203935 TI - Treatment of Helicobacter pylori infection favourably affects gastric mucosal superoxide dismutases. AB - BACKGROUND AND AIMS: Excessive production of reactive oxygen metabolites (ROMs) by phagocytic cells is thought to contribute to the mucosal pathology of Helicobacter pylori infection. Previously, H pylori infection was shown to have a differential effect on some gastric mucosal scavenger enzymes of ROMs-namely, mitochondrial and cytoplasmic superoxide dismutases-reflected by a large increase in the cytokine inducible manganese superoxide dismutase and a marginal decrease in the constitutive copper/zinc superoxide dismutase. The present study was performed to evaluate whether these altered mucosal superoxide dismutase concentrations and activities in H pylori associated gastritis are reversed to normal by successful treatment of the infection. PATIENTS AND METHODS: In two different treatment groups-namely, omeprazole or ranitidine, in combination with clarithromycin and metronidazole (OME/AB (n = 33) and RAN/AB (n = 30))-manganese superoxide dismutase and copper/zinc superoxide dismutase concentrations were evaluated by enzyme linked immunosorbent assays in homogenates of gastric antrum and corpus biopsy specimens obtained before and eight weeks after successful treatment of H pylori infection. Superoxide dismutase activities in these homogenates were determined spectrophotometrically in eight patients of both groups before and after successful treatment. The concentrations of gastric mucosal superoxide dismutases were also determined in 12 patients with a persistent H pylori infection, with (n = 4) or without (n = 8) eradication therapy. Infection and eradication of H pylori were confirmed by a combination of culture and histology. RESULTS: Concentrations of manganese superoxide dismutase were significantly lower after than before therapy in antral (p < 0.001 in both treatment groups) and corpus (p < 0.001 in both treatment groups) mucosa. By contrast, copper/zinc superoxide dismutase concentrations were significantly higher (p < 0.001) only in antral mucosa of the OME/AB treated group. Manganese superoxide dismutase activity was significantly lower after than before treatment in antral (OME/AB p < 0.01, RAN/AB p < 0.001), but not in corpus mucosa. Copper/zinc superoxide dismutase activity was not significantly altered by therapy. In the 12 patients with a persistent H pylori infection no major changes in the gastric mucosal superoxide dismutase concentrations were found. CONCLUSIONS: The raised manganese superoxide dismutase and reduced copper/ zinc superoxide dismutase concentrations and activities in H pylori associated gastritis were reversed towards normal by successful treatment of the infection. PMID- 9203936 TI - The inhibitory effect of glucagon-like peptide-1 (GLP-1) 7-36 amide on gastric acid secretion in humans depends on an intact vagal innervation. AB - BACKGROUND: Glucagon-like peptide-1 (GLP-1)(7-36) amide is an intestinal incretin hormone which also inhibits gastric acid secretion in humans. Its mechanism of action is unclear, but it strongly inhibits vagally induced secretion (sham feeding), suggesting that it could influence vagal activity. AIM/METHODS: The effect of intravenous GLP-1 (7-36 amide) (1 pmol/kg/min) was studied on pentagastrin induced acid secretion in otherwise healthy subjects, previously vagotomised for duodenal ulcer (n = 8) and in a group of young (n = 8) and old (n = 6) healthy volunteers. RESULTS: Pentagastrin increased acid secretion significantly in all three groups, but the plateau concentration in the vagotomised subjects was lower than in controls. Infusion of GLP-1 (7-36 amide) significantly inhibited acid secretion in the control groups (to 67 (SEM 6) and 74 (SEM 3)% of plateau concentrations in young and old controls, respectively) but had no effect in the vagotomised subjects. Differences in plasma concentrations of GLP-1 (7-36 amide), recovery of gastric marker, duodenal regurgitation, or Helicobacter pylori status could not explain the lack of effect. Blood glucose was lowered equally by GLP-1 (7-36 amide) in all subjects. CONCLUSION: The inhibitory effect of GLP-1 (7-36 amide) on acid secretion depends on intact vagal innervation of the stomach. PMID- 9203937 TI - L-arginine in low concentration improves rat intestinal water and sodium absorption from oral rehydration solutions. AB - BACKGROUND: The nitric oxide (NO) precursor L-arginine has been shown to produce variable effects on intestinal absorptive function, including ion transport. AIMS: To determine whether there is an optimal concentration of L-arginine, promoting proabsorptive effects from oral rehydration solutions (ORS) with 90 or 60 mM sodium. SUBJECTS AND METHODS: In vivo perfusion of rat jejunum with determination of net water absorption, unidirectional fluid exchanges, sodium and calcium transport, and glucose absorption. RESULTS: L-Arginine (1 mM) added to the 90 mM sodium ORS increased intestinal absorption of both sodium and water. Higher concentrations of L-arginine (2 to 10 mM) lacked this stimulatory effect. At 20 mM, L-arginine decreased sodium absorption below baseline. With a 60 mM sodium ORS, 2 mM L-arginine had a maximal fluid and electrolyte proabsorptive effect. At 20 mM L-arginine, net water absorption was indistinguishable from that obtained in the absence of L-arginine, and lower than with 2 mM L-arginine. Sodium absorption remained raised above baseline in perfusions with 10 and 20 mM L-arginine. Morphologically, villi from perfusions with increased absorption showed a large expansion of intercellular and lamina propria intercellular spaces. CONCLUSIONS: Low concentrations of L-arginine seem to stimulate water and electrolyte absorption by the small intestine. This effect is consistent with NO induced vasodilation, may be vaso-constrictive and thereby reverse fluid and electrolyte transport. PMID- 9203938 TI - Intestinal tolerability of nitroxybutyl-flurbiprofen in rats. AB - BACKGROUND: Nitric oxide derivatives of non-steroidal anti-inflammatory drugs (NSAIDs) are thought to be much less ulcerogenic than their parent compounds. AIM: To compare the effect and potency of flurbiprofen and nitroxybutyl flurbiprofen to uncouple mitochondrial oxidative phosphorylation (an early pathogenic event in NSAID enteropathy), increase intestinal permeability (transitional stage), and cause macroscopic small intestinal damage. METHODS: In vitro uncoupling potency was assessed using isolated coupled rat liver mitochondria and in vivo by electron microscopy of rat small intestinal mucosa (two hours after the drugs). A dose-response study with flurbiprofen (single doses of 5, 10, 20, and 40 mg/kg) and equimolar doses of nitroxybutyl flurbiprofen was performed; assessing their effect on intestinal permeability (at 18-20 hours), with 51Cr EDTA, and the number of pointed (< 5 mm) and longitudinal (> 5 mm) small intestinal ulcers at 24 hours. RESULTS: Flurbiprofen, but not nitroxybutyl-flurbiprofen, stimulated coupled respiration in vitro. Both drugs, however, uncoupled in vivo; in the case of nitroxybutyl-flurbiprofen possibly because hydrolysis of its ester bond released free flurbiprofen. Intestinal permeability was uniformly and equally increased with both drugs compared with controls. The number of small intestinal ulcers, pointed and longitudinal, was significantly reduced with nitroxybutyl-flurbiprofen apart from the number of longitudinal ulcers with the highest dose. CONCLUSIONS: These studies show that nitroxybutyl-flurbiprofen is associated with significantly less macroscopic damage in the small intestine than flurbiprofen but was associated with mitochondrial damage in vivo and caused similar increases in permeability of the small intestine, suggesting that its beneficial effect is on the later pathogenic stages of the damage. PMID- 9203939 TI - Daytime and night time motor activity of the small bowel after solid meals of different caloric value in humans. AB - BACKGROUND: Meals disrupt the interdigestive pattern of small bowel motor activity and convert it into the postprandial pattern. Previous studies have shown that duration of postprandial motor activity depends on the caloric value of a meal, but results from two recent human studies suggested that there is a caloric ceiling, above which an additional increase in the caloric load fails to prolong the postprandial period further. AIM: To investigate the hypothesis of a caloric ceiling by studying daytime motor activity of the human small bowel in response to five solid meals, covering a wide range of calories. METHODS: Eight healthy male volunteers underwent five separate, ambulatory small bowel manometry studies and had a total of 80 meals. For lunch, volunteers ate between one and five portions of a solid meal (220, 440, 660, 880, or 1100 kcal). Ten hours later and 30 minutes before they went to bed, they ate either two or four portions of the same meal (440 kcal or 880 kcal). Recordings were analysed visually for phase III of the migrating motor complex and a validated computer program calculated incidence and amplitude of contractions. RESULTS: Apart from two versus three portions (440 kcal v 660 kcal), postprandial motor activity was significantly prolonged by each 220 kcal increase in the caloric load of the lunch (168 (SEM 14), 305 (22), 298 (23), 368 (36), and 398 (38) min). Mean incidence of contractions was significantly different only between the two extremes tested: 220 kcal and 1100 kcal (2.9 (0.3) v 4.5 (0.6) min-1). Amplitude of contractions did not depend on meal size. Daytime and night time postprandial activity were not significantly different. This was true for duration of fed activity, as well as mean incidence and amplitude of contractions during the postprandial period. CONCLUSION: Caloric value of a meal regulates duration of the fed activity in the human small bowel over a wide range of calories, and-for caloric loads up to 1100 kcal-there is no maximum duration of postprandial motor activity. Furthermore the postprandial small bowel motor activity is very similar between daytime and night time. PMID- 9203940 TI - Non-steroidal anti-inflammatory drugs are associated with emergency admission to hospital for colitis due to inflammatory bowel disease. AB - BACKGROUND: To evaluate the relation between non-steroidal anti-inflammatory drugs (NSAIDs) and colitis due to inflammatory bowel disease. METHODS: A case control study was conducted using a prospectively constructed, record linkage database containing hospital event and dispensed drug data (1989-93). The study population consisted of 319,465 people resident in Tayside in January 1989, and still resident (or dead) in October 1994. RESULTS: Of the 785 patients admitted to hospital as emergencies with colitis between July 1989 and June 1993, 200 fulfilled the case criterion of colitis due to inflammatory bowel disease. A further 1198 persons were used as community controls. Odds ratios were calculated for three exposure periods (current, recent, and past exposure). The overall odds ratios (with 95% confidence intervals) for current and recent exposure to NSAIDs were 1.77 (1.01 to 3.10) and 1.93 (1.20 to 3.09) respectively. Current and recent exposure to NSAIDs was also associated for incident cases, with odds ratios of 2.96 (1.32 to 6.64) and 2.51 (1.13 to 5.55). There was a trend for recent exposure among non-incident cases. CONCLUSION: The use of NSAIDs may be associated with an increased risk of emergency admission to hospital for colitis due to inflammatory bowel disease, particularly among patients with no previous history. PMID- 9203941 TI - Lack of association between an interleukin-1 receptor antagonist gene polymorphism and ulcerative colitis. AB - BACKGROUND: Recently, the association of a polymorphism in the gene coding for the anti-inflammatory cytokine interleukin-1 receptor antagonist with ulcerative colitis has been reported. This was interpreted as a possible genetic predisposition for severity of the inflammatory response. AIMS: To examine this polymorphism in a southern German population. SUBJECTS: The study included 234 healthy controls, 57 patients with ulcerative colitis, including 31 patients with pancolitis, 44 first degree healthy relatives of patients with ulcerative colitis, and 65 patients with Crohn's disease. METHODS: Genotypes were determined by a polymerase chain reaction amplification of the intron 2 fragment harbouring a variable number of tandem repeat nucleotide sequences. Amplification products were separated on a 2% agarose gel. RESULTS: The allele frequency for allele 2 was 27% in healthy controls, 28% in Crohn's disease, and 21% in patients with ulcerative colitis. The same allele frequency (21%) was found in a subgroup of patients with ulcerative colitis affecting the whole colon. Thus for allele 2 as well as for all other alleles, genotypes, or carriage rates no significant differences were found compared with controls. All allele frequencies in the control population were similar to those in earlier studies. CONCLUSIONS: No association of a polymorphism in the interleukin-1 receptor antagonist gene with ulcerative colitis could be identified in this southern German population. The findings of an earlier study reporting an increased frequency of allele 2, particularly in patients with pancolitis, could not be confirmed. PMID- 9203942 TI - Treatment of ulcerative colitis in the cottontop tamarin using antibody to tumour necrosis factor alpha. AB - BACKGROUND: The aetiology and pathophysiology of ulcerative colitis remains unclear; however, there is increasing recognition of the critical role of inflammatory cytokines in the pathogenesis of this disease. Among these, tumour necrosis factor alpha (TNF alpha) seems to play an important role. AIM: To study the effects of an engineered human monoclonal antibody to TNF alpha (CDP571) in the treatment of idiopathic ulcerative colitis in the cottontop tamarin. METHODS: Six cottontop tamarins with confirmed ulcerative colitis received repeated doses of CDP571. Progression of disease was assessed by measuring both body weight and rectal biopsy pathology. RESULTS: All animals showed a rapid improvement in clinical condition and rectal biopsy pathology that was maintained following completion of the therapy. CONCLUSION: These studies indicate the efficacy of selective antibody therapy to TNF alpha for the treatment of ulcerative colitis in a primate and suggest that similar therapy in human could be of value. PMID- 9203943 TI - Changes in barrier function of a model intestinal epithelium by intraepithelial lymphocytes require new protein synthesis by epithelial cells. AB - BACKGROUND: Elements of the mucosal immune system may play an important part in regulating epithelial barrier function in the intestinal tract. Intraepithelial lymphocytes (IELs) represent a subtype of immunocyte which is strategically placed to regulate epithelial function at most mucosal sites. AIMS AND METHODS: An IEL derived cell line (SC1) was used to examine its effects on the model epithelium T84--a tumour derived cell line which retains the phenotype of colonic crypt cells. Transepithelial electrical resistance (TER) was used as a marker of epithelial integrity. RESULTS: Coculture of T84 cells with SC1 produced a significant fall in TER as did exposure of T84 monolayers to IEL derived supernatant. Recombinant interferon-gamma (rIFN gamma) also reduced TER in T84 monolayers. Cycloheximide prevented the effects of IEL supernatant and of rIFN gamma on TER. The fall in TER in response to rIFN gamma was attenuated by blocking antibodies, which did not alter the fall in resistance induced by IEL supernatant. Fractions of IEL supernatant, separated on the basis of size, evoked temporally distinct changes in TER. Ultrastructural studies support the hypothesis that the slow onset but severe fall in TER indicates catastrophic effects on the monolayer. The more rapid onset fall in TER was not associated with gross changes in monolayer morphology. Reduction of TER by IEL supernatant was not influenced by inhibitors of tyrosine phosphatase or of protein kinase C. Although herbimycin did reduce the rapid onset change in TER, the tyrosine kinase inhibitor genistein did not alter responses to IEL supernatant. CONCLUSIONS: Mucosal T cells may influence barrier function by a process involving new protein synthesis by epithelial cells. This model may have relevance in some inflammatory conditions of the gastrointestinal tract. PMID- 9203944 TI - Gastrointestinal motor mechanisms in hyperglycaemia induced delayed gastric emptying in type I diabetes mellitus. AB - BACKGROUND: Hyperglycaemia delays gastric emptying, both in healthy controls and in patients with diabetes mellitus. The effect of hyperglycaemia on antroduodenal motility in diabetes has not yet been studied. AIM: To investigate the gastrointestinal motor mechanisms involved in the hyperglycaemia induced retardation of gastric emptying in patients with type I diabetes mellitus and autonomic neuropathy. In eight diabetic patients antroduodenal manometry was performed simultaneously with scintigraphic measurement of emptying of a mixed solid-liquid meal, during euglycaemia (5-8 mmol/l glucose) and hyperglycaemia (16 19 mmol/l glucose), on separate days, in random order. RESULTS: Hyperglycaemia decreased the cumulative antral motility index from 38.3 (range 24.2-47.6) to 30.8 (range 17.3-38.1) (p = 0.025) and reduced the number of antral pressure waves propagated over > or = 4.5 cm (p = 0.04). Duodenal phase III-like activity was seen irrespective of the glycaemic state (in three patients during euglycaemia and in four patients during hyperglycaemia). Hyperglycaemia significantly affected gastric emptying of the solid meal: it prolonged the lag phase from 20.0 minutes to 28.5 minutes (P = 0.02), increased the 50% emptying time from 73.5 minutes to 104.5 minutes (p = 0.03), and increased the percentage of isotope remaining in the stomach after 120 minutes from 33.5% to 46.5% (p = 0.02). The cumulative antral motility index was correlated with the 50% emptying time (r = 0.75, p = 0.02) during euglycaemia, but not during hyperglycaemia (r = 0.28, P = 0.31). Liquid emptying was not influenced by the blood glucose concentration. CONCLUSIONS: Hyperglycaemia reduces postprandial antral contractile activity and its organisation in patients with type I diabetes and autonomic neuropathy. These changes in antroduodenal motility are likely to constitute the mechanism through which gastric emptying of solids is delayed during high blood glucose concentrations in these diabetic patients. PMID- 9203945 TI - Clinical significance of serum p53 antigen in patients with pancreatic carcinomas. AB - BACKGROUND: Alterations in the p53 gene are often found in pancreatic cancer, and accumulation of the p53 protein has been noted in tumour cells. AIMS: To investigate whether serum p53 protein concentrations could be used as markers for p53 gene mutations in neoplasms of the pancreas. METHODS: Serum p53 protein concentrations were determined by an enzyme linked immunosorbent assay (ELISA) in 104 cases of pancreatic adenocarcinoma, and 61 matched formalin fixed tissue sections were also stained by an anti-p53 DO-7 monoclonal antibody. RESULTS: The mean serum concentration of p53 protein in the adenocarcinoma patients was 0.27 (SEM 0.02) ng/ml, and was significantly higher than in 35 healthy blood donors (0.15 (0.02) ng/ml, SD = 0.11) or in 15 cases of chronic pancreatitis (0.15 (0.02) ng/ml). Adopting an arbitrary cut off value for the serum p53 protein concentration of 0.37 ng/ml, which corresponded to a value 2 SD above the mean value from the healthy blood donors, positive serum p53 protein concentrations were found in 23 out of 104 (22.1%) patients with adenocarcinomas examined, 16 out of 47 (34.0%) patients with carcinomas with distant metastases, but only seven of 57 patients (12.3%) with carcinomas without metastases (p < 0.05). In 11 patients with pancreatic adenocarcinomas, the mean serum p53 protein concentration after tumour resection was 0.21 (0.05) ng/ml, and had decreased compared with the preoperative concentrations (0.25 (0.05) ng/ml) (P < 0.05). There were no significant associations between the serum concentrations of p53 protein and serum concentrations of markers such as CA19-9 or CEA; however, serum concentrations of p53 protein demonstrated a potential role as an additional tumour marker. Immunohistochemical studies disclosed that the p53 protein was expressed in 28 out of 61 pancreatic adenocarcinomas (45.9%). Serum p53 protein concentrations in the positively immunostained cases were significantly higher than in the negatively immunostained cases (0.35 (0.05) ng/ml v 0.15 (0.01) ng/ml; p < 0.005). Furthermore, positive immunostaining for p53 protein was found in eight out of 10 (80%) serum positive p53 protein cases with adenocarcinomas. CONCLUSION: An increase in serum p53 protein concentrations appears during the progression of pancreatic adenocarcinoma and correlates with the accumulation of p53 protein as a result of a mutation of the p53 gene. An analysis of p53 antigen concentrations can detect p53 gene alterations, which could be useful for the selection of treatment regimens. PMID- 9203946 TI - Allele loss, replication errors and loss of expression of E-cadherin in colorectal cancers. AB - BACKGROUND: Loss of E-cadherin expression has been implicated in the development of invasive characteristics in colorectal carcinomas. Failure to express E cadherin may result from mutations of the E-cadherin gene (HSECAD). AIMS: To examine the correlation between E-cadherin expression and genetic changes at HSECAD; and to examine differences in E-cadherin expression and genetic changes at HSECAD between three different groups of colorectal cancer--replication error positive (RER+) sporadic cancers, RER--sporadic cancers and ulcerative colitis associated cancers. SUBJECTS AND METHODS: Sixty eight colorectal cancers (22 RER+ sporadic cancers, 32 RER- sporadic cancers and 14 ulcerative colitis associated cancers) were studied using immunohistochemistry and for allele loss at the HSECAD locus. Exon 16 of HSECAD contains several mononucleotide repeat tracts which are very similar to microsatellite repeats and which may be susceptible to replication errors (manifest as new alleles). All cases were also examined for new alleles in exon 16. RESULTS: Absent or decreased E-cadherin protein expression was found in 27 (38%) of 68 colorectal cancers and the pattern of expression did not differ significantly among the three tumour groups. Allele loss occurred at HSECAD in four (10%) of 40 informative cancers and there were no differences between the three subgroups. New alleles at exon 16 were detected in three (14%) of 22 RER+ tumours; no new alleles were detected in RER- or ulcerative colitis associated cancers. Overall, there was no correlation between allelic loss or exon 16 replication errors and immunohistochemical E-cadherin expression. CONCLUSIONS: (1) Loss of E-cadherin expression probably does not occur as a result of mutation at the HSECAD locus in colorectal cancers. (2) There is no difference in the frequency of loss of heterozygosity at HSECAD among RER+, RER- and ulcerative colitis associated colorectal cancers. PMID- 9203948 TI - Reduction in renal blood flow following acute increase in the portal pressure: evidence for the existence of a hepatorenal reflex in man? AB - BACKGROUND: To investigate the relation between changes in portal haemodynamics and renal blood flow (RBF) in patients with cirrhosis. PATIENTS/METHODS: Twenty patients with cirrhosis and transjugular intrahepatic portosystemic stent-shunts were divided into two groups which were well matched. At routine portography, either changes in unilateral RBF (group I) or changes in cardiac output (group II) before and after shunt occlusion were studied. Blood was obtained from the renal and systemic circulations for the measurement of neurohumoral factors before and after shunt occlusion in group I patients. RESULTS: After shunt occlusion, there was a progressive reduction in unilateral RBF from a mean (SD) of 289 (32) to 155 (25) (-43.5%) (p < 0.001). These changes correlated significantly with the changes in the portal atrial gradient (p < 0.001). There was no significant change in heart rate, mean arterial pressure and right atrial pressure. No significant changes were found in the concentrations of the various neurohumoral factors measured. There was a less notable but significant reduction in the cardiac output (-10.9%) (p = 0.02) unaccompanied by significant reduction in the pulmonary capillary wedge pressure or mean arterial pressure. CONCLUSIONS: These results suggest the existence of hepatorenal reflex in man which is important in the regulation of RBF, although other mechanisms may also be contributory. PMID- 9203947 TI - High frequency of K-ras mutations in human colorectal hyperplastic polyps. AB - BACKGROUND: Hyperplastic polyps are common benign colorectal polyps, and are thought to have little association with malignant tumours in the colorectum. However, several reports suggest that some hyperplastic polyps may develop into colorectal neoplasms. AIM: To clarify genetic alterations in colorectal hyperplastic polyps. METHODS: Twenty eight colorectal polyps having serrated components were resected from patients endoscopically. The K-ras gene mutations in codons 12 and 13 were analysed by PCR-RFLP. Intranuclear p53 protein was immunostained by the avidin-biotin complex method. RESULTS: A mutation of the K ras gene was detected in nine (47%) of 19 hyperplastic polyps, and five (56%) of nine adenomas. p53 protein nuclear accumulation was detected immunohistochemically in two (22%) of nine adenomas, but not in any of the hyperplastic polyps. CONCLUSION: Some hyperplastic polyps may be true neoplastic lesions, and could be precursors of malignant neoplasia. PMID- 9203949 TI - A prospective evaluation of cytology from biliary strictures. AB - BACKGROUND: Bile duct strictures may be benign or malignant. A definite diagnosis is desirable to advise patients of their prognosis and to identify any amenable to curative surgery. AIMS: To compare different methods of cytology sampling from biliary strictures and evaluate the use of cytology in this context. PATIENTS AND METHODS: In a prospective study 54 patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) had cytology samples obtained as follows: (1) biliary stricture brushings, (2) from the screw thread of a "Soehendra stent retriever" inserted through the stricture, (3) from the proximal end of a blocked biliary stent, and (4) cellular material spun down from a 20 ml specimen of bile. Examination of slides and rinsings was performed by an expert cytologist who graded them for the adequacy of the sample and for evidence of malignancy. RESULTS: Prolonged follow up disclosed malignancy in 52 of the 54 cases, the other two being chronic pancreatitis. Bile samples provided adequate cytology samples in 44%, the Soehendra stent retriever in 70%, retrieved stents in 84%, and cytology brush sampling in 96%. Overall, 28 malignancies were detected by cytology, including 14 of 28 pancreatic carcinomas and 12 of 16 cholangiocarcinomas. Twenty two of the malignancies were detected by brush sampling and the other methods added a total of another six cases. CONCLUSIONS: Cytology sampling is best done by brushing the biliary stricture. Cytology sampling can confirm the diagnosis in 75% of cholangiocarcinomas and 50% of pancreatic carcinomas. The techniques involved are simple to perform and should be routine clinical practice whenever potentially malignant biliary strictures are encountered at ERCP. PMID- 9203950 TI - Fluoroscopically guided laser lithotripsy versus extracorporeal shock wave lithotripsy for retained bile duct stones: a prospective randomised study. AB - BACKGROUND AND AIMS: To compare extracorporeal shock wave lithotripsy (ESWL) and laser induced shock wave lithotripsy (LISL) of retained bile duct stones to stone free rate, number of therapeutic sessions, and costs. PATIENTS: Thirty four patients were randomly assigned to either ESWL or LISL therapy. The main reasons for failure of standard endoscopy were due to stone impaction (n = 12), biliary stricture (n = 8), or large stone diameter (n = 14). METHODS: An extracorporeal piezoelectic lithotripter with ultrasonic guidance and a rhodamine 6G laser with an integrated stone tissue detection system were used. LISL was performed exclusively under radiological control. RESULTS: Using the initial methods complete stone fragmentation was achieved in nine of 17 patients (52.4%) of the ESWL group and in 14 of 17 patients (82.4%) in the LISL group, or combined with additional fragmentation techniques 31 of the 34 patients (91.2%) were stone free at the end of treatment. In comparison LISL tended to be more efficient in clearing the bile ducts (p = 0.07, NS). Significantly less fragmentation sessions (1.29 v 2.82; p = 0.0001) and less additional endoscopic sessions (0.65 v 1.6; p = 0.002) were necessary in the LISL group. There were no major complications in either procedure. CONCLUSIONS: Compared with ESWL, fluoroscopically guided LISL achieves stone disintegration more rapidly and with significantly less treatment sessions, which leads to a significant reduction in cost. PMID- 9203951 TI - Molecular analysis of T cell receptor beta variability in a patient with orofacial granulomatosis. AB - BACKGROUND: Orofacial granulomatosis (OFG) is a rare chronic inflammatory disorder of unknown causation and is characterised histologically by non caseating granulomas and aggregates of small lymphocytes. The molecular nature of these T cells is, however, unclear. AIMS: To determine the T cell receptor (TCR) V beta gene usage of the T cell infiltrate associated with the primary lesions in a patient with OFG. METHODS: A molecular method involving reverse transcriptase (RT)-polymerase chain reaction (PCR), DNA cloning, single strand conformation polymorphism (SSCP), length analysis, and nucleotide sequencing was used. RESULTS: Compared with peripheral blood lymphocytes from the same patient, notably restricted TCRV beta gene usage was observed in the T cell infiltrate. Only three of the 24 major TCRV beta gene families were represented in the repertoire. There was preferential usage of the V beta 6 gene. In addition, more than 20% of the V beta 6 TCR transcripts exhibited an identical unique V-D-J junctional sequence, suggesting a local antigen driven V beta 6 T cell clonal expansion in vivo, a phenomenon not observed in normal oral mucosa. CONCLUSIONS: The TCRV beta repertoire of T cells associated with OFG is restricted. It is also associated with a local T cell clonal expansion. The results, therefore, provide a new perspective on the immunopathology of OFG. PMID- 9203952 TI - "Golf ball liver": agent orange hepatitis. AB - 2,4-Dichlorophenoxyacetic acid (2,4-D) is a selective weedkiller which works by uncoupling oxidative phosphorylation and is in widespread use. It is known as "agent orange". A 65 year old man had acute hepatitis, thought to be caused by exposure to 2,4-D. The patient ingested 2,4-D as a result of habitual licking of his golf ball. Clinical and histological data together with a challenge test confirmed the diagnosis of "golf ball liver". PMID- 9203953 TI - Familial hyperamylasaemia. AB - A six year old boy underwent extensive investigation for recurrent abdominal pain and was found to have a persistently raised serum amylase. Endoscopic retrograde cholangiopancreatography was normal and macroamylasaemia was excluded. Serum amylase concentrations were found to be raised in other family members spanning three generations, all of whom were asymptomatic. Clearance studies suggested no evidence of a renal tubular defect and serum lipase concentrations were normal. This is the first report of apparently familial hyperamylasaemia and the mode of inheritance is consistent with an autosomal dominant pattern. PMID- 9203954 TI - Metaplastic (hyperplastic) polyps of the large bowel: benign neoplasms after all? PMID- 9203955 TI - Human oesophageal bicarbonate secretion: a phenomenon waiting for a role. PMID- 9203956 TI - Pigbel-like syndrome in a vegetarian in Oxford. PMID- 9203957 TI - Bone disease after liver transplantation should not be underestimated. PMID- 9203958 TI - Characterization of the bovine C alpha gene. AB - The complete genomic sequence of a bovine C alpha gene is reported here. The genomic sequence was obtained from a C alpha phage clone that had been cloned from a genomic EMBL4 phage vector library. The C alpha sequence had previously been expressed as a chimeric antibody and identified as IgA using IgA-specific antibodies. Intron/exon boundaries were determined by comparison of the genomic sequence with an expressed bovine C alpha sequence obtained from spleen by reverse transcription-polymerase chain reaction (RT-PCR). Analysis of 50 Swedish bovine genomic DNA samples using genomic blots and five different restriction enzymes failed to detect evidence of polymorphism. However, PstI digests of Brown Swiss DNA showed a restriction fragment length polymorphism (RFLP), suggesting that at least two allelic variants of bovine IgA exist. Comparison of the deduced amino acid sequence of bovine IgA with sequences available for other species indicated that the highest homology was with that of swine, another artiodactyl. This was the highest homology observed for all mammalian IgA compared except for that between IgA1 and IgA2 in humans. Bovine IgA shares with rabbit IgA3 and IgA4, an additional N-linked glycosylation site at position 282. However, the collective data indicate that cattle are like swine and rodents and unlike rabbits in having a single locus of the gene encoding IgA of this species. PMID- 9203959 TI - The serum resistance of malaria-infected erythrocytes. AB - IgG and IgM antibodies were detected on non-parasitized as well as parasitized erythrocytes (E) from mice surviving over 15 days after infection with rodent malaria, Plasmodium berghei, whereas C3 was detected exclusively on parasitized E. Parasitized E, however, were quite resistant to the haemolytic activity of guinea pig complement and effectively inactivated human C3b to iC3b on their surface. Similarly, parasitized E were extremely resistant to homologous complement as assessed by haemolysis and C3 binding even when regulatory proteins (decay-accelerating factor, DAF; complement receptor related gene y, Crry; heat stable antigen, HSA) were blocked with specific antibodies. DAF and Crry were equally expressed on both normal E and parasitized E from mice within a week post infection; therefore, molecules that inhibit the haemolysis or C3 binding of parasitized E appear to be independent of DAF and Crry. Unexpectedly, the molecular forms of HSA and DAF in parasitized erythrocyte membranes were found to be different from those of normal erythrocyte membranes: DAF was detected as three bands (85,000, 64,000 and 30,000 MW) by immunoblotting. HSA was detected as more highly glycosylated forms than normal HSA. These alterations of DAF and HSA could be explained by the modification of membrane proteins and polysaccharides induced by parasitization, and we hypothesize that these changes of membranes or membrane proteins are involved in the resistance of parasitized E against homologous complement. PMID- 9203960 TI - Obtaining a functional recombinant anti-rhesus (D) antibody using the baculovirus insect cell expression system. AB - The cloning and production of a human anti-rhesus (Rh) D monoclonal antibody (mAb) using the baculovirus-insect cell expression system is described. This monoclonal recombinant antibody R.D7C2 derived from a human parental IgM lambda immunoglobulin was obtained after immortalization of lymphocytes by Epstein-Barr virus (EBV). The human heavy (VH) and light (VL) variable regions were cloned from the parental cell line and genetically fused to the human constant IgG1 heavy (H) and light (L) chain genes (gamma 1 and lambda, respectively). A recombinant baculovirus was constructed that directs the co-expression of genes encoding both genetically fused heavy and light chains under the control of two late and strong baculovirus promoters. After infecting the Spodoptera frugiperda (Sf9) insect cell line with this baculovector, a complete IgG1 mAb was secreted in the culture medium indicating that each immunoglobulin chain was correctly processed and assembled with a functional glycosylation into a tetrameric form. In vitro analysis showed that the functional properties of R.D7C2 using agglutination tests were efficient for the specific recognition of Rh-D-positive red blood cells (RBC). In addition, R.D7C2 showed effector functions of the gamma 1 heavy chain resulting in the lysis of Rh+ papain RBC by an antibody-directed cellular cytotoxicity mechanism. These results demonstrate that R.D7C2 can be produced in the baculovirus-insect cell expression system as a source for potential therapeutic application in the treatment of the haemolytic disease of the newborn. PMID- 9203961 TI - Mutation of recombinant complement component C9 reveals the significance of the N terminal region for polymerization. AB - Complement component C9 binds to C5b-8 sites on target cells and polymerizes to form the membrane attack complex (MAC). The aim of the work reported here was to discover which region within C9 was responsible for protecting the globular protein against self-polymerization. Computer prediction modelling highlighted the domain at the N-terminus of C9, which was then investigated by site-directed mutagenesis. The mutated proteins were expressed using insect cells infected with baculovirus. Removal of 16, 20 or 23 amino acids at the N-terminus of C9 resulted in inactivation due to self-polymerization. In contrast, removal of 4, 8 or 12 amino acids resulted in a C9 that did not polymerize spontaneously, had two to threefold enhanced lytic activity on erythrocytes, and had increased binding to C5b-8 sites on rat neutrophils. These results suggest that the domain within the first 16 amino acids at the N-terminus of C9 is crucial in preventing the self polymerization of the globular protein. We have also found that C9 contains a motif (27WSEWS31) common to a family of cytokine receptors that is similar to a tryptophan-rich motif (WEWWR) of the membrane pore formers, thiol-activated cytolysins. Mutation of this motif in C9 resulted in polymerized protein, consistent with this site keeping the N-terminus in a protected conformation and preventing premature self-polymerization. PMID- 9203962 TI - Altered interleukin-2 receptor alpha-chain is expressed in human T-cell leukaemia virus type-I-infected T-cell lines and human peripheral blood mononuclear cells of adult T-cell leukaemia patients through an alternative splicing mechanism. AB - A polymerase chain reaction (PCR) method was used to detect the interleukin-2 receptor alpha-chain (IL-2R alpha) chain which lacks the conventional transmembrane (TM) domain in mRNA from human T-cell leukaemia virus type-I (HTLV I)-infected cell lines or peripheral blood mononuclear cells (PBMC) isolated from adult T-cell leukaemia (ATL) patients. Primer pairs encompassing the TM domain were selected to generate a 357-base pair (bp) fragment. A 146-bp PCR product was observed consistently in addition to the target 357-bp PCR product in mRNA from HTLV-I-infected cell lines, such as MT-1, MT-2, MT-4 and in PBMC isolated from ATL patients. However, this 146-bp PCR product was undetectable in HTLV-I negative cell lines. The product was also detected in PBMC from normal individuals if activated in vitro with phytohaemagglutinin but not without stimulation. DNA sequence analyses revealed that exons from 5 to 7, which define a 211-bp region containing the conventional TM domain, were deleted in the 146-bp PCR product. The C-terminal amino acid sequence starting from Gly174 of the 211 bp-deleted molecule was distinct from that of conventional IL-2R alpha as a result of an altered reading frame. We identified a 45000 MW peptide generated from IL-2R alpha mRNA through this exon skip in cell lysate of MT-1 and MT-2 by Western blot analyses using an antibody raised against the peptides specific to an altered IL-2R alpha. Our results indicate that an altered IL-2R alpha chain is expressed in HTLV-I-infected T lymphocytic cell lines and in ATL patients. PMID- 9203963 TI - Interleukin-7 in the skin of Schistosoma mansoni-infected mice is associated with a decrease in interferon-gamma production and leads to an aggravation of the disease. AB - The effect of recombinant interleukin-7 (rIL-7) on the course of murine schistosomiasis and the development of the accompanying immune response were investigated. We demonstrated that IL-7 expression could be detected in the skin of infected mice from 1 to 21 days following infection. We here report that intradermal injection of exogenous human IL-7, prior to the penetration of the parasite into the skin, leads to a more severe liver pathology and an increased number of surviving adult parasites. In addition, injection of rIL-7 alters parasite migration (estimation of burdens of young larvae in lungs and liver). Administration of rIL-7 led to a decrease of IL-12 and interferon-gamma-(IFN gamma) specific messengers RNA in skin and, more markedly, in skin-draining lymph nodes. The number of B220 expressing cells was increased, and T-cell number was reduced, in IL-7-treated infected mice. In addition, levels of IFN-gamma and IL-4 in sera were significantly reduced, whereas there was a shift from a Th1 to a Th2 type associated humoral response towards the egg antigens. Our experimental observation illustrate that the exogenous administration of rIL-7 affects both the development of the host's immune response and the behaviour of the parasite within the infected host. The early and specific production of IL-7 in the host skin, following infection with Schistosoma mansoni, raises fascinating questions concerning the relationships between the parasite and its host at the very beginning of their interaction. PMID- 9203964 TI - The roles of tumour necrosis factor-alpha, interleukin-1 and interleukin-12 in murine cytomegalovirus infection. AB - The roles of the inflammatory cytokines tumour necrosis factor-alpha (TNF-alpha), interleukin-1 (IL-1) and IL-12, in murine cytomegalovirus (MCMV) disease were investigated in susceptible BALB/c and resistant C57BL/6 mice. MCMV infection induced IL-1 and TNF-alpha production by peritoneal cells from BALB/c mice, as demonstrated previously in C57BL/6 mice. Overt ill-health and viral replication in the spleens of BALB/c mice were increased by in vivo treatment with soluble TNF-alpha receptors to inhibit the activity of this cytokine, whilst antibodies to IL-12 had a similar but more restricted effect C57BL/6 mice were not affected by either treatment, suggesting TNF-alpha and IL-12 are not critical for natural killer cell-mediated restriction of viral replication in the spleen. Soluble TNF alpha receptors and antibodies to IL-12 also enhanced MCMV titres and numbers of viral antigen-positive cells in the livers of BALB/c mice and TNF-alpha receptors have similar effects in C57BL/6 livers. In contrast, IL-1 receptors improved the health of MCMV-infected BALB/c mice and reduced viral replication and hepatitis at some time-points. Mechanisms which may underlie these changes are discussed. PMID- 9203965 TI - IL-13 production by allergen-stimulated T cells is increased in allergic disease and associated with IL-5 but not IFN-gamma expression. AB - Interleukin-13 (IL-13) shares many, but not all, of the properties of the prototypic T-helper type 2 (Th2) cytokine IL-4, but its role in allergen-driven T cell responses remains poorly defined. We hypothesized that allergen stimulation of peripheral blood T cells from patients with atopic disease compared with non atopic controls results in elevated IL-13 synthesis in the context of a 'Th2 type' pattern. Freshly isolated peripheral blood mononuclear cells (PBMC) obtained from sensitized atopic patients with allergic disease, and non-atopic control subjects, were cultured with the allergens Phleum pratense (Timothy grass pollen) or Dermatophagoides pteronyssinus (house dust mite) and the non allergenic recall antigen Mycobacterium tuberculosis purified protein derivative (PPD). Supernatant concentrations of IL-13, along with IL-5 and interferon-gamma (IFN-gamma) (Th2- and Th1-type cytokines, respectively) were determined by enzyme linked immunosorbent assay (ELISA). Allergen-induced IL-13 and IL-5 production by T cells from patients with allergic disease was markedly elevated (P = 0.0075 and P = 0.0004, respectively) compared with non-atopic controls, whereas IFN-gamma production was not significantly different. In contrast to allergen, the prototypic Th1-type antigen M. tuberculosis PPD induced an excess of IFN-gamma over IL-13 and IL-5 production, and absolute concentrations of cytokines were not affected by the presence or absence of atopic disease. Addition of exogenous recombinant IFN-gamma or IL-12, cytokines known to inhibit Th2-type responses, significantly inhibited allergen-driven production of both IL-13 and IL-5, but not T-cell proliferation, whereas exogenous IL-4 did not significantly affect production of IL-13 or IL-5. We conclude that allergen-specific T cells from atopic subjects secrete elevated quantities of IL-13 compared with non-atopic controls, in the context of a Th2-type pattern of cytokine production. PMID- 9203966 TI - TCR gamma delta+ cells are prominent in normal bovine skin and express a diverse repertoire of antigen receptors. AB - More than 80% of T cells in bovine skin localized in the superficial 0.5 mm of the dermis. Only 3% occurred within the epidermis or made contact with the stratum basale while the remainder occupied deeper dermal sites. The gamma delta T-cell receptor (TCR) was expressed by 44% of T cells in skin and 39% and 35% expressed, respectively, the CD4 and CD8 markers. Some cells co-expressed CD8 and the gamma delta TCR. A highly diverse repertoire of gamma delta TCR was expressed in skin due mainly to the usage of multiple V delta segments and to extensive sequence variation at the junctions of both TCR gamma and TCR delta chains. However, a single receptor isotype was used. Transcripts encoding several new components of the bovine gamma delta TCR were identified, including three new V gamma segments, the C gamma 5 region and 13 new functional V delta segments. Taken together with earlier findings, these results emphasize that ruminant gamma delta T cells express exceptionally diverse antigen receptors and suggest they may have a more elaborate recognitive capacity than do their counterparts in other species. PMID- 9203967 TI - Reconstitution of SCID mice with haemopoietic precursors: a detailed analysis of gamma delta T-cell reconstitution. AB - A well-known characteristic of gamma delta T cells is that they are produced in waves during ontogeny, with cells expressing T-cell receptor V gamma 5 appearing early in fetal thymic ontogeny, followed by V gamma 6, then by other gamma delta T-cell types. In addition, evidence exists to suggest that the potential of haemopoietic precursors to generate different types of gamma delta T cells changes in ontogeny. We have used these observations as the basis for an extensive study of the potential for haemopoietic precursors isolated from fetal liver, neonatal spleen and adult bone marrow to reconstitute severe combined immunodeficient (SCID) mice. Mice that were reconstituted as newborns with fetal liver cells most closely resembled normal C.B-17 mice with respect to both lymphocyte numbers and subsets, while mice reconstituted with adult bone marrow had fewer cells than normal mice. This deficit spanned both T and B cells in all organs examined. Among the gamma delta T-cell subsets examined, the ability to reconstitute V gamma 4+ cells was particularly dependent on the ontogenic age of the reconstituting presursors, with fetal liver cells having the greatest capacity to generate V gamma 4+ cells, and adult bone marrow cells the least. The vast majority of the T cells produced in the reconstituted mice were of donor origin, and the level of reconstitution was found to be dependent upon some factor other than the presursor frequency. PMID- 9203968 TI - Characterization of the mucosal cell-mediated immune response in IL-2 knockout mice before and after the onset of colitis. AB - One of the major advances in the understanding of inflammatory bowel disease has been the observation that mice with immunoregulatory defects, such as interleukin 2 knockout (IL-2 -/-) mice, develop spontaneous gut inflammation. Here we have characterized the immune response in the ileum, caecum and colon of these mice before and after the onset of colitis by examining the cellular infiltrate, the cytokines produced by these cells and the mucosal vascular addressin MAdCAM-1. IL 2 -/- mice developed colitis after 35 days of age and before this the mice were apparently healthy. IL-2 -/- mice aged over 35 days with colitis had large numbers of CD4+, CD8+, alpha beta T-cell receptor (TCR)+ and gamma delta TCR+ T cells, macrophages, dendritic cells and MAdCAM-1+ endothelial cells in the caecum and colon. This was associated with an increase in the number of interferon-gamma (IFN-gamma), IL-1 and tumour necrosis factor-alpha (TNF-alpha) transcripts and a decrease in IL-4 and IL-10 transcripts. Treatment of IL-2 -/- mice with cyclosporin A significantly delayed mortality. Interestingly, IL-2 -/- mice under 35 days, although healthy, did show some subtle immunological signs of preclinical disease. There was a significant increase in the number of macrophages and dendritic cells in the colonic lamina propria and increased mRNA for IL-1 and TNF-alpha. There were also increased numbers of MAdCAM-1+ endothelial cells, but IFN-gamma transcripts were not elevated. These results suggest that T-cell-mediated colitis in IL-2 -/- mice may be secondary to an initial non-specific inflammation. PMID- 9203969 TI - Impaired calcium mobilization and differential tyrosine phosphorylation in intestinal intraepithelial lymphocytes. AB - Intestinal intraepithelial lymphocytes (iIEL) exhibit a unique activation state characterized by the expression of activation markers and effector functions, but a minimal response to mitogenic signals in vitro. To further characterize this activation status, iIEL were compared with splenic T cells for two key activation signals, calcium mobilization and tyrosine phosphorylation. Calcium mobilization was impaired in iIEL treated with the calcium ionophores ionomycin or A23187, thapsigargin, or by CD3-cross-linking. The calcium mobilization defect is shared by mature and embryonic iIEL. Anti-phosphotyrosine Western blot analysis revealed that the iIEL are able to respond to T-cell receptor (TCR)-mediated signals by tyrosine phosphorylation, although the patterns of phosphorylation differ from those seen in splenic T cells. We conclude that iIEL are unable to mobilize calcium in vitro, which may be due to modulation of TCR-mediated signal transduction pathways by the microenvironment of the intestinal epithelium and/or caused by the standard isolation procedure used to prepare iIEL, which must be considered in future in vitro studies of iIEL function. PMID- 9203970 TI - Self-reactive forbidden clones are confined to pathways of intermediate T-cell receptor cell differentiation even under immunosuppressive conditions. AB - It is believed that self-reactive forbidden T-cell clones are generated by 'failure' of the pathway of T-cell differentiation in the thymus, if it is disturbed. We examined how such forbidden clones are generated under immunosuppressive conditions. Mice were treated with an injection of deoxyspergualin, FK506, or cycloporin A. From day 3, the number of cells yielded by various organs decreased. Because of the resistance of intermediate (int) T cell receptor (TCR) cells (i.e. TCRint cells), they became more prominent in proportion than TCRhigh cells. TCRhigh cells are conventional T cells generated through the mainstream in the thymus, whereas TCRint cells are primordial T cells generated by the extrathymic pathway or an alternative intrathymic pathway. Similar to untreated mice, forbidden V beta 3+ and V beta 11+ clones in C3H/He (Mls-1b2a) mice were confined to TCRint cells after treatment; there was no leakage of forbidden clones into TCRhigh cells in the thymus and periphery. In parallel with the increase in the proportion of TCRint cells, the proportion of forbidden clones also increased under immunosuppressive states, especially in the liver. Liver mononuclear cells isolated from treated mice still had the potential to mediate autologous killing. The present results suggest that the generation of self-reactive clones is highly restricted to the pathways of TCRint cell differentiation even under immunosuppressive conditions. PMID- 9203971 TI - CD45 enhances positive selection and is expressed at a high level in large, cycling, positively selected CD4+CD8+ thymocytes. AB - T-cell development is arrested at the CD4+CD8+ (DP; double-positive) stage of thymocyte development in CD45 null mice. However, the mechanism by which CD45 participates in the positive selection of T cells remains to be investigated. In this report we describe a DP thymocyte population that associates positive selection with expression of high levels of CD45, CD4 and CD8. DP thymocytes of this phenotype are large, cycling cells and represent approximately 20% of DP thymocytes in normal mice. In mice expressing a transgenic T-cell receptor (TCR) specific for the male antigen presented by H-2Db (H-Y TCR), the up-regulation of TCR, CD5 and CD69 in this large DP population occurred in a major histocompatibility complex (MHC)-restricted manner. To investigate further the role of CD45 in positive selection, we determined whether thymocytes that expressed a transgenic CD45RO molecule under the control of the proximal lck promoter can influence the positive selection of T cells in H-Y TCR transgenic mice. It was found that in female H-Y TCR transgenic mice, MHC-restricted positive selection of CD4- CD8+ H-Y TCR+ thymocytes was enhanced by increased CD45RO expression. Thus, CD45 increases the efficacy of positive selection of CD4 CD8+ thymocytes that express H-Y TCR. PMID- 9203972 TI - Two distinct pathways of human macrophage differentiation are mediated by interferon-gamma and interleukin-10. AB - Forming cellular conjugates with T cells, macrophages can help their targets to mount an immune response or they can destroy the targeted T cell. The two functions are performed by two distinct macrophage subsets that can be distinguished by cell surface marker phenotypes, B7+ CD16- and B7- CD16+. Interferon-gamma (IFN-gamma) induces the former, interleukin-10 (IL-10) induces the latter phenotype. The two macrophage differentiation pathways are mutually exclusive; each cytokine inhibits the effect of the other cytokine. The second messenger cAMP enhances the macrophage B7 expression and suppresses the macrophage CD16 expression. However, together with IL-10, cAMP blocks the generation of both macrophage phenotypes. In the chimpanzee we noted deviations from this differentiation pattern that are suggestive of an enhanced IL-10 presence in the primate environment. PMID- 9203973 TI - A role for interferon-gamma and transforming growth factor-beta in erythroid cell mediated regulation of nitric oxide production in macrophages. AB - Interferon-gamma (IFN-gamma) and transforming growth factor-beta (TGF-beta) are known to be a potent inducer and inhibitor for macrophage (Mo) activation process, respectively. In the present study we established that the nucleated erythroid cells (NEC) separated from the spleens of adult (CBA x C57BL/6)F1 (CBF, H-2k/H-2d) mice following phenylhydrazine treatment are potentially capable of inducing nitric oxide (NO) production in thioglycollate broth-elicited peritoneal macrophages (Mo). The stimulating effect of both NEC and their culture supernatant on NO secretion by Mo was most apparent in the presence of bacterial lipopolysaccharide (LPS) and neutralizing antibodies (Abs) to TGF-beta and was largely reversed by the addition to the culture of neutralizing Abs to IFN-gamma. Collectively these results suggest that NEC, through production of IFN-gamma and TGF-beta, may exert a regulatory influence on development and functionality of cells pertaining to monocyte (Mc)/Mo lineage. PMID- 9203974 TI - Granulocyte-macrophage colony-stimulating factor elevates invariant chain expression in immature myelomonocytic cell lines. AB - Invariant chain (Ii) plays an important role in major histocompatibility complex (MHC) class II antigen processing and presentation and is constitutively synthesized in B lymphocytes, in macrophages, dendritic cells and in some epithelial cells. It has been shown that interferon-gamma, tumour necrosis factor alpha and interleukin-4 co-regulate Ii and MHC class II expression in various cell types. We describe here a novel regulation of Ii expression in macrophages. Treatment of the premature monocytic cell lines WEHI 265.1, M1 and WEHI-3B with recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) strongly enhances Ii expression while class II expression is not induced. In contrast, GM CSF did not enhance Ii in mature macrophage cell lines. The increase of Ii expression in WEHI 265.1 cells takes several days. This long induction time, and a difference in activity between GM-CSF-conditioned medium and GM-CSF, together suggest that GM-CSF stimulates WEHI 265.1 cells to secrete a factor that modulates Ii expression. These results may imply a class II-independent function of Ii, which we discuss in this paper. PMID- 9203975 TI - Enhancement of B-cell translocation gene-1 expression by prostaglandin E2 in macrophages and the relationship to proliferation. AB - Although prostaglandin (PG) E2 is known to suppress various macrophage functions, the molecular mechanisms by which that occurs are largely unknown. To understand better those mechanisms, differential screening of a cDNA library from PGE2 treated macrophages was performed. Subsequently, the DNA sequence of a differentially expressed cDNA clone was determined and the cDNA was identified as B-cell translocation gene-1 (BTG1), a recently cloned antiproliferative gene. A two-to threefold increase in macrophage BTG1 expression was observed after PGE2 treatment. PGE1 and platelet-activating factor, but not leukotrienes B4, and C4, or lipopolysaccharide, also enhanced BTG1 expression. Furthermore, this effect ws mimicked by dibutyryl cAMP which indicated the involvement of elevated cAMP in the PGE2-mediated enhancement of BTG1. Moreover, there was an inverse correlation between BTG1 mRNA expression and macrophage proliferation; however, BTG1 alteration was not associated with macrophage tumoricidal activation. Thus, BTG1 may play a role in PGE2-mediated inhibition of macrophage proliferation and not activation. PMID- 9203976 TI - Importance of combined treatment with IL-10 and IL-4, but not IL-13, for inhibition of monocyte release of the Ca(2+)-binding protein MRP8/14. AB - Expression of the two myeloic related proteins MRP8 and MRP14 is restricted to distinct stages of monocytic differentiation. Heterodimeric MRP8/14 complexes (27E10 antigen) have been shown to represent their biologically active forms. In this study, we investigated the effects of Th2-cytokines on release of these proteins from freshly obtained blood monocytes and monocytes cultured for 7 days in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Monocytes were stimulated with pokeweed mitogen (PWM) in the presence or absence of interleukin-13 (IL-13), IL-4 and IL-10, and secretion of MRP8, MRP14 and MRP8/14 was assessed by using a sandwich enzyme-linked immunosorbent assay system. Peripheral monocytes secreted significantly increased amounts of MRP14 and MRP8/14 but not MRP8 under stimulation with PWM. IL-10 and IL-4, but not IL 13, down-regulated the PWM-stimulated MRP8/14 secretion in a dose-dependent manner. Maximal inhibition required that IL-10 and IL-4 be added up to 1 h before or simultaneous with PWM. A combination of IL-10 and IL-4 even at suboptimal concentrations significantly suppressed protein secretion much more than using IL 10 or IL-4 at a doubled concentration alone. Peripheral monocytes cultured for 7 days in the presence of GM-CSF showed two-to threefold higher protein levels compared with freshly obtained blood monocytes but responded inefficiently to either IL-4, IL-13, or IL-10 alone. However, treatment with IL-10 in combination with IL-4 but not IL-13 strongly suppressed MRP14 and MRP8/14 release by these cells. The unresponsiveness of 7-day-cultured blood macrophages suggests that more differentiated and activated cells may lose their ability to respond to anti inflammatory cytokines. Combined cytokine treatment may therefore more effectively control the progression of chronic inflammatory processes. PMID- 9203977 TI - Generation of dendritic cell-like antigen-presenting cells in long-term mixed leucocyte culture: phenotypic and functional studies. AB - The mechanisms contributing to the proliferation and differentiation of antigen presenting cell (APC) precursors upon antigen stimulation or tissue injury are poorly understood. Herein, we report the induction of a population of dendritic cell-like cells (DLC) with potent antigen-presentation function from unfractionated spleen cells by means of repetitive allostimulation in long-term mixed leucocyte cultures (LT-MLC). Initially, only a few adherent DLC were observed. By 4-6 weeks, however, there were large numbers of DLC which survived persistently. Features of these DLC are closely related to dendritic cells (DC), including (1) dendritic, veiled or spiny-processed morphology; (2) expression of a wide array of leucocyte surface markers including DC-associated or restricted antigens: 33D1, NLDC-145, CD11c (N418), heat-stable antigen (HSA), CD44, B7-1 and B7-2; (3) ability to migrate to draining lymph nodes and white pulp area of spleen; (4) expression of high level of major histocompatability complex (MHC) class II molecules and (5) more potent mixed leucocyte reaction (MLR)-stimulating capacity than peritoneal macrophages and APC-enriched spleen cells. DLC stimulated MLR was inhibited by monoclonal antibodies (mAbs) to B7-1, B7-2, intracellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), leucocyte-function associated antigen-1 (LFA-1) or very-late activation antigen-4 (VLA-4) by 30-55%. When maintained for more than 2 months, the DLC did not lose their MLR-stimulating activity, but many surface markers were down regulated except for Mac-2 and VCAM-1, which remained stable or were up regulated, respectively. In short-term culture, the addition of granulocyte macrophage colony-stimulating factor (GM-CSF) or interleukin (IL)-2 enhanced proliferation of DLC, while tumour necrosis factor-alpha (TNF-alpha) and IL-4 did not. IL-4 suppressed not only 'spontaneous', but also GM-CSF-enhanced proliferation, suggesting that cytokines play a differential role in DLC proliferation. These results confirm that professional APC can proliferate in response to repetitive antigen stimulation, and their proliferation is differentially regulated by cytokines. A comparison study of DLC with typical DC is being carried out in our laboratory. PMID- 9203978 TI - Steroids inhibit uptake and/or processing but not presentation of antigen by airway dendritic cells. AB - Recent studies from our laboratory indicate that local and (particularly) systemic steroids can modulate the traffic of dendritic cells (DC) through resting and inflamed airway epithelial tissues. The present report focuses upon the T-cell activating properties of DC, which are controlled by granulocyte macrophage colony-stimulating factor (GM-CSF) signals, and in particular the question of whether the DC-stimulating effects of GM-CSF are susceptible to regulation by steroids. We present evidence that while dexamethasone inhibited GM CSF-dependent uptake and/or processing of exogenous antigen by DC, it was ineffective in blocking the presentation of preprocessed self antigen to alloreactive T cells in a one-way mixed lymphocyte reaction (MLR). Associated GM CSF-induced up-regulation of major histocompatability complex (MHC) class II and CTLA4 ligand expression by DC were also unaffected by dexamethasone phosphate (DX), reinforcing the view that the inhibitory effects of steroids on the T-cell activating functions of DC are restricted to steps upstream from presentation of processed antigen to the T-cell receptor (TCR). These findings have potentially important implications in relation to the use of topical steroids in the treatment of atopic asthma, a disease in which local T-cell activation in airway tissue is a key pathogenic factor, and which furthermore is characterized by intense production of GM-CSF within the airway epithelium. PMID- 9203979 TI - Induction of the 2B9 antigen/dipeptidyl peptidase IV/CD26 on human natural killer cells by IL-2, IL-12 or IL-15. AB - Activation of human natural killer (NK) cells involves sequential events including cytokine production and induction of cell surface molecules, resulting in the enhancement of cytolytic activity. To delineate the activation process of NK cells, we generated murine monoclonal antibodies (mAbs) against YT, a human large granular lymphocyte/natural killer (LGL/NK) cell line. Among the mAbs reactive with YT cells, one mAb, termed 2B9, was noted because of the lack of reactivity with most of the human T- and B-cell lines tested. In fresh peripheral blood mononuclear cells (PBMC), however, the majority of cells expressing this antigen (Ag) were T cells but not CD16+ nor CD56+ NK cells. Since YT cells showed an activated phenotype expressing interleukin-2 (IL-2) receptor alpha chain, we examined whether 2B9 Ag could be induced on normal human peripheral blood NK cells by cytokines known to activate NK cells. The 2B9 Ag was induced on NK cells by IL-2, IL-12 or IL-15 while no induction was observed by interferon-gamma (IFN gamma). Biochemical analysis showed that anti-2B9 mAb recognized a 115 kDa molecule in YT cells. A cDNA clone encoding the 2B9 Ag was isolated from a cDNA expression library of YT cells and its sequence was identical to CD26 cDNA although it was not of full length. Transient expression of the 2B9 cDNA on COS-7 cells revealed that this cDNA encodes the antigenic epitope(s) recognized by anti 2B9 mAb as well as Ta1, an anti-CD26 mAb. These results showed that the 2B9 Ag is identical to CD26, and demonstrated that CD26 is an activation antigen on CD16+ CD56+ NK cells inducible by IL-2, IL-12 or IL-15. PMID- 9203980 TI - Regulation of angiogenesis by hypoxic stress: from solid tumours to the ovarian follicle. AB - The preovulatory follicle provides a unique physiological example of rapid growth accompanied by neovascularization, two processes that are generally characteristic of pathologies such as wound repair or malignancy. During the hours preceding ovulation, follicular growth is accompanied by elevated levels of messenger RNA for vascular endothelial growth factor (VEGF). Angiogenic activity, mediated by VEGF, is manifested in the peripheral blood vessels surrounding the follicle, that show capillary sprouting and increased vascular permeability. Following ovulation, rapid infiltration of capillaries through the follicular wall is essential for the formation of the corpus luteum. In this review we compare the preovulatory follicle with a popular model of avascular solid tumour growth, namely the multicellular tumour spheroid, in particular the role of hypoxic stress in the regulation of angiogenesis in both systems. PMID- 9203981 TI - Injury-induced alterations in newly synthesized sulphated proteoglycans from rabbit arterial neointima covered by regenerated endothelium. AB - Proteoglycan (PG) synthesis is a highly regulated, dynamic process that is known to be altered during atherogenesis. Endothelial injury, which may be the primary event in atherosclerosis, has been reported to stimulate PG synthesis and accumulation in the arterial extracellular matrix. The objective of this investigation was to study injury-induced alterations in PG synthesis and accumulation in the neointima, developed in response to a selective balloon catheter de-endothelialization of aortas of normocholesterolaemic rabbits. One group of rabbit aortas was incubated with 35S-Na2SO4 for 8 h, to study in vitro the de novo synthesis of sulphated PG. Another group of rabbit aortas was used to study PG accumulated in aortic neointima vs. PG present in intima-media of normal aortas. Newly synthesized sulphated PG was characterized by light microscopic radioautography and size exclusion chromatography. Purified intimal-medial PG extracts from unlabelled aortas were analysed for protein and glycosaminoglycan (GAG) content and GAG distribution pattern. Results from this study revealed that the neointima of injured aortas synthesized sulphated PG at a significantly higher concentration than the intima of normal aortas. Size exclusion chromatography revealed that neointima synthesized higher molecular weight PG, and in a higher proportion, than its counterpart PG from normal aortas. PG accumulated in neointima of injured aortas showed a significantly altered GAG distribution pattern. These data confirm that neointima developed in response to injury synthesizes and accumulates PG which is altered compared to PG present in the intima-media of normal aorta. PMID- 9203982 TI - Experimental cutaneous leishmaniasis: transmission electron microscopy of the inoculation site. AB - Tissue response against inoculation of Leishmania (Leishmania) amazonensis promastigotes in the hind footpad was quite different between two strains of mice: in BALB/c animals there was parasitism of perineurial cells by the 8th week post inoculation (WPI) and heavy parasitism of macrophages, as well as degenerated extracellular parasites close to collagen fibers at the 39th WPI, whereas in C57BI/6j mice there was heavy parasitism of macrophages at 6th WPI, dermal vessels with high endothelial cell at 21st WPI and well preserved intracellular amastigote forms by 51st WPI. In both animals there was no parasitism of keratinocytes or Langerhans cells. Thus BALB/c mice were useful as an experimental model for diffuse cutaneous leishmaniasis and showing a new feature, parasitism of perineurial cells, whereas C57BI/6J animals show hypersensitivity signs, together with a few preserved parasites, only late in the course of infection. From a morphological point of view, there were no differences in macrophages, or in the interaction between this target cell and the parasite, between the animal models studied. This suggests that the difference in the response of the hosts towards the parasite could depend on the way in which they activate a cellular, i.e. lymphocyte mediated immune, response. PMID- 9203983 TI - Regeneration of myoepithelial cells in rat submandibular glands after yttrium aluminium garnett laser irradiation. AB - The regeneration of myoepithelial cells in rat submandibular salivary gland after partial irradiation with yttrium aluminium garnett (YAG) laser was investigated. The irradiated glands were examined immunohistochemically for actin, histochemically for alkaline phosphatase (ALP), and by transmission electron microscopy (TEM). In control glands, myoepithelial cells were positive for actin and ALP. Electron microscopically, the positive reaction for actin was associated with the myofilaments of myoepithelial cells, and the plasma membrane of myoepithelial cells was positive for ALP. One day after YAG laser irradiation, the irradiated region was necrotic. By 5 days, duct-like structures and epithelial clusters were observed at the interface between the necrotic zone and the remaining undamaged glands; immature acini appeared after 7 days. No reaction in duct-like structures or epithelial clusters to actin or ALP was recognizable by 5 days. However, at 7 days, actin and ALP-positive spindle cells appeared at the periphery of the duct-like structures and immature acini. After 10 days, both actin-positive and ALP-positive cells increased in number. These observations indicate that during regeneration, actin-positive and ALP-positive cells regenerate myoepithelial cells, and it is suggested that this differentiation to myoepithelial cells is closely related to that of luminal to acinar cells. In addition, TEM observations indicate that regenerated myoepithelial cells originated from the basal cells of duct-like structures. PMID- 9203984 TI - An age-related change in susceptibility of rat brain to encephalomyocarditis virus infection. AB - Rats were inoculated intraperitoneally (i.p.) or intracerebrally (i.c.) with 1 x 10(4) plaque forming units (PFU)/animal of the D variant of encephalomyocarditis virus (EMC-D) at 2, 4, 7, 14, 28 or 56 days of age for virological and histopathological examination. In the i.p.-inoculation study, neither viral replication nor lesions were detected in the animals inoculated at 28 and 56 days of age. In the animals inoculated when younger than 14 days of age, lesions were restricted to the brain although viral replication was detected in the brain, heart and pancreas. The brain lesions were characterized by acute meningoencephalitis with neuronal necrosis in the cerebral cortex, hippocampus and thalamus, and viral RNA was detected in degenerated and/or intact neurons. In the i.c.-inoculation study, similar age-related changes in susceptibility of rat brain to EMC-D infection were observed, but a minor difference was that viral replication and lesions were still detected in the hippocampus of some animals inoculated at 28 days of age. These results suggest that an age-related decrease in the susceptibility of rat brain to EMC virus infection may reflect an age related change in the susceptibility of neurons themselves as well as in maturation of the immune system. PMID- 9203985 TI - Bile secretion by the rat liver during synchronized regeneration. AB - Simultaneous coexistence of differentiated, proliferating and redifferentiated hepatocytes occurs during normal liver regeneration (LR). The aim of the present work was to study the time course of the capacity of the liver to form bile during synchronized LR. Following two-thirds partial hepatectomy (PH) in rats, i.v. administration of the ribonucleotide reductase reversible inhibitor hydroxyurea (HU) was used to transiently block liver cells at G1/S boundary. Experiments were performed at 0 and 4 hours, and 1, 3 or 7 days after releasing HU-induced inhibition. Bile acid pool size was determined by collecting bile samples over 24 hours. Initial (first hour) bile flow and bile acid output were increased early on during synchronized LR as compared with the values found in non-hepatectomized control animals. These values were thereafter (1 day) reduced, but increased again at 3 days after halting HU infusion. The time course of bile acid depletion and changes in bile flow were very similar in control and synchronized LR, except that in the latter a more important early reduction in bile flow and bile acid output was found. Shortly after PH, part of the bile acid pool was lost, but this was quickly restored, soon (1 day) reaching a net bile acid pool size very similar to that found in control rats. The highest pool size relative to liver weight was found on day 1, when bile acid output and bile flow reached their lowest values. Additional experiments were performed using in situ perfused regenerating rat livers in which stepwise infusion of taurocholate (TC) was carried out. PH alone modified neither the bile acid-independent (BAIF) nor the bile acid-dependent fraction of bile flow (BADF). However, in normal LR, the BAIF decreased on day 1 and recovered at 7 days, while in synchronized LR it remained depressed up to 7 days. The BADF was only reduced during the early phase of normal LR and did not change significantly in synchronized LR. The maximal secretion rate (SRmax) for TC, as expressed per gram of remaining liver tissue, was not affected immediately after PH, but a marked reduction was observed on day 1 in both normal and synchronized LR. Afterwards, SRmax was quickly restored in both synchronized LR and, although in a slower way, normal LR. These results suggest that synchronization of LR involves changes in the time required to the recovery of specific liver functions such as bile formation. PMID- 9203986 TI - Synthesis of glycoconjugates by human diseased veins: modulation by procyanidolic oligomers. AB - Venous diseases become steadily more common and severe with age, and are often accompanied by venous lymphatic oedema. We have investigated the role of glycoconjugates in this disorder and the action of procyanidols used to treat these diseases. Explants of vein wall from patients with or without venous lymphatic edema were cultured for 24 hours and the incorporation of radioactive glucosamine into total glycoconjugates and into hyaluronan was measured. The explants from patients with oedema incorporated more glucosamine than those without oedema (+42% expressed as c.p.m./mg dry weight into total glycosaminoglycans and +12% expressed as c.p.m./mg dry weight into hyaluronan). The explants from oedematous patients secreted less glycoconjugates into the culture medium than those from non-oedematous veins (-63% of total incorporated radioactivity into hyaluronan and -66% into hyaluronidase-resistant glycoconjugates). Explants placed in medium containing procyanidols (1 mg/ml, 2.8 mM) incorporated less glucosamine (-19%) and secreted more into the medium (+119%). Glycoprotein and sulphated glycosaminoglycan synthesis were mainly affected which may well explain the beneficial effect of procyanidols on vein disorders. PMID- 9203987 TI - Stem cell factor and c-kit in the mammalian testis: lessons originating from Mother Nature's gene knockouts. AB - Analysis of mice with naturally occurring mutation in the genes encoding the tyrosine kinase receptor protein, c-kit, and its ligand, stem cell factor (SCF), has implicated these proteins in testicular development and function because of the phenotype of these mice of reduced or absent fertility. Descriptive and functional studies have highlighted the involvement of these proteins in primordial germ cell migration and survival, and in spermatogonial proliferation, survival and adhesion. This review summarises the current knowledge regarding functions of c-kit and SCF in the testis and discusses the implications of information gleaned from other biological systems. PMID- 9203988 TI - Regulation of the natriuretic peptide system in rat uterus during the estrous cycle. AB - Uterine natriuretic peptides may be involved in the alterations that occur in the uterus during the estrous cycle through its role in hydromineral balance. The following studies were performed to determine whether uterine natriuretic peptides and receptors follow a cyclic pattern during the estrous cycle. The results obtained show that atrial natriuretic peptide (ANP) content in rat uterine tissue was low in proestrus (8.5 +/- 2.6 pg/g) and significantly increased (P < 0.001) in estrus (71.5 +/- 16.6 pg/g), metestrus (82.6 +/- 19.7 pg/g) and diestrus (91.0 +/- 19.4 pg/g), whereas plasma ANP was not altered during the cycle. Similarly, measurement of uterine ANP mRNA by reverse transcribed polymerase chain reaction (RT-PCR) indicated lowest levels of ANP mRNA at proestrus. Measurement of C-type natriuretic peptide (CNP) by a specific and sensitive radioimmunoassay revealed that uterine CNP also varies with the estrous cycle. Uterine CNP was low in diestrus (143.2 +/- 22.4 pg/mg protein) as compared with proestrus, estrus and metestrus (305.3 +/- 51.5, 267.5 +/- 44.9, 291 +/- 41.2 pg/mg protein respectively, P < 0.05). Autoradiography performed on uterine tissue slices localized natriuretic peptide receptors to myometrial smooth muscle layers and to endometrial uterine glands. High binding of 125I-ANP was observed in proestrus and estrus with 60-75% decreases during metestrus and diestrus. Binding of 125I-tyr0CNP to uterine slices was also high during proestrus, but declined by 35% at estrus, metestrus and diestrus. The alterations in the receptors were also observed at the level of synthesis. RT-PCR detection of guanylyl cyclase A (GC-A) receptor mRNA and guanylyl cyclase B (GC-B) mRNA showed high signals at proestrus but 4- and 2-fold reductions respectively at metestrus and diestrus. In conclusion, variations in uterine ANP and CNP and their receptors during the rat estrous cycle imply the involvement of the natriuretic peptides in uterine hydromineral balance and myometrial motor activity. PMID- 9203989 TI - Dietary essential fatty acid supplementation, urinary calcium excretion and reproductive performance in the diabetic pregnant rat. AB - Hypercalciuria may be a contributory factor to the disturbed calcium homoeostasis seen in diabetic pregnant rats and their offspring. In diabetes, essential fatty acid metabolism is impaired. We have therefore investigated whether feeding a diet supplemented with essential fatty acids will ameliorate the hypercalciuria of diabetic pregnancy and improve reproductive performance. Female rats were fed a standard rat diet, a fat-free diet plus evening primrose oil or a fat-free diet plus sunflower oil. They were injected with streptozotocin or vehicle and mated. Urine samples were analysed for calcium before injection and during gestation. Term-pregnant diabetic rats fed evening primrose oil showed a 73% reduction in urinary calcium output compared with similar rats fed standard diet (P < 0.001). The corresponding reduction was 44% in diabetic rats fed sunflower oil (P < 0.001). A depletion of essential fatty acids in diabetes may therefore be associated with hypercalciuria; dietary supplementation, particularly with evening primrose oil, appears to correct the problem. Diabetic pregnant rats fed evening primrose oil showed a significantly greater live fetal mass (85 +/- 2 vs 33 +/- 12 g; P < 0.05) compared with similar rats fed standard diet. Such findings may imply a normalization of placental transport by essential fatty acids. Rats fed evening primrose, but not sunflower oil, also showed a reduced incidence of diabetes after streptozotocin injection compared with rats fed standard diet (63 vs 86%). Rats fed on evening primrose oil that did become diabetic were less hyperglycaemic than those on the standard diet (29 +/- 2 vs 37 +/- 2 mmol/l), suggesting that the oil may have anti-diabetic properties. PMID- 9203990 TI - Prolactin receptors in human meningiomas: characterization and biological role. AB - Sixty cerebral meningioma specimens obtained at surgery from 34 female and 26 male patients were examined for the presence of prolactin (PRL) receptors. These were compared with normal arachnoid tissue from which these tumours arise. PRL receptors were detected in 61.7% of meningiomas whereas no PRL binding was found in samples of normal arachnoid tissue. No relationship was found when sex or histological findings were compared with the presence of PRL receptors. Receptor positive tumours had saturable and high-affinity (Kd, 4.8 +/- 0.5 ng/ml) receptors with hormonal specificity for human PRL (hPRL) resembling that of other target tissues of PRL in man. The biological role of these receptors was investigated in primary cell cultures derived from meningioma tissue characterized for PRL receptor. When human PRL was added to the culture medium, in doses ranging from 1 to 200 ng/ml, a dose-dependent stimulation of 3H thymidine incorporation was observed only in PRL-receptor positive tumours. The PRL concentrations required to produce a half-maximal effect ranged from 11 to 20 ng/ml and were quite close to the dissociation constant (Kd) of binding of PRL to its receptors. PRL also caused an increase of cell number compared with control with a significant effect after 3 and 4 days of culture. In conclusion, these findings indicate that a large number of human meningiomas express specific and functional receptors for PRL which are involved in mediating its proliferative effects. PMID- 9203991 TI - Rabbit sex hormone binding globulin: primary structure, tissue expression, and structure/function analyses by expression in Escherichia coli. AB - Sex hormone binding globulin (SHBG) is a homodimeric plasma protein found in mammals that binds sex steroids with high affinity and regulates their bioavailability. The protein is identical in structure and properties to the androgen binding protein (ABP) found in the male reproductive tract. We have isolated a 1245-base pair rabbit SHBG cDNA encoding a reading frame for a signal peptide followed by a protein of 367 amino acids, which shares 79.0, 68.1 and 63.2% amino acid identity with the corresponding human, rat and mouse proteins respectively. Northern blot and hot-nested PCR analyses indicated that rabbit SHBG is produced from a 1.6 kilobase mRNA in the liver of both sexes and in the testis. The rabbit SHBG cDNA was inserted into pGEX-1 lambda T for expression of a glutathione S-transferase/SHBG fusion protein in Escherichia coli. The bacterial product bound 5 alpha-dihydrotestosterone (DHT) in the same manner as the corresponding protein in serum. The dissociation constants (Kd) for rabbit and human SHBGs produced in E. coli were 11.1 +/- 1.1 nM and 2.1 +/- 0.6 nM respectively, and rabbit SHBG formed a less stable protein-steroid complex (t1/2 = 5 min) than human SHBG (t1/2 > 60 min). Unlike human SHBG, rabbit SHBG does not bind estradiol with high affinity. To aid in the identification of differences in the sequences of rabbit and human SHBG, which determine species differences in steroid-binding affinity and specificity, chimeras containing the 5'-terminal half of SHBG from one species and 3'-terminal half of SHBG from the other species were constructed and expressed. It was found that the chimeric proteins assumed similar steroid-binding affinity and specificity as the wild-type proteins when the amino (N)-terminal half of SHBG was derived from the same species. Replacement of the carboxyl (C)-terminal half of rabbit SHBG by the corresponding region of the human molecule increased the integrity of its steroid-protein complex. This supports the concept that amino acids within the N-terminal half of SHBG constitute the steroid-binding domain while the C-terminal half of the molecule may provide structural stability to the protein and its steroid-binding site. PMID- 9203992 TI - Pituitary and gonadal responses to the long-term pulsatile administration of gonadotrophin-releasing hormone in fetal sheep. AB - The fetal hypothalamo-pituitary-gonadal axis reaches a peak in activity at mid gestation and this is followed by a period of suppression which persists until the onset of puberty. The decline in gonadotrophic activity during late gestation is thought to reflect the maturation of central and peripheral feedback signals. In order to establish if sustained pituitary responsiveness is rate limiting to the reinstatement of reproductive function, we have examined the endocrine consequences of repeated pulsatile GnRH administration to male and fetal sheep during late gestation. Beginning on day 121 of gestation (term = 145 days) chronically catheterized fetal sheep were given i.v. pulses of either 500 ng GnRH or saline every 2 h for 14 days. Pituitary and gonadal responses were assessed by measuring changes in plasma concentrations of LH, FSH, inhibin and testosterone (in male fetuses) in response to the first pulse of GnRH on day 1 and to the corresponding pulse on days 4, 7, 10 and 14. In response to the first pulse of GnRH there was an immediate release of LH, with the peak response being significantly (P < 0.01) greater than on subsequent days. In male fetuses each pulse of LH was followed by a rise in plasma testosterone concentrations within 40-60 min. The amplitude of these testosterone responses increased significantly (P < 0.01) after 9 days of treatment despite a decline in the plasma LH response. Basal FSH concentrations increased progressively (P < 0.05) during pituitary stimulation with GnRH in both male and female fetuses. Immunoreactive inhibin concentrations were significantly (P < 0.05) higher in males than in females, and there was a gradual increase throughout the experimental period irrespective of treatment. We observed no inverse correlation between inhibin and FSH concentrations. These data show that pulsatile administration of GnRH to fetal sheep during late gestation results in sustained re-activation of pituitary gonadal function. The decline in fetal gonadotrophins, which is a characteristic feature of late gestation, is therefore likely to result from inadequate GnRH secretion from the fetal hypothalamus rather than an inhibition of pituitary function by peripheral feedback signals. PMID- 9203993 TI - Effect of pregnancy on rat myometrial beta 2-adrenoceptor mRNA and isoproterenol induced relaxation of isolated uterine strips. AB - The altered myometrial contractile activity near term of pregnancy is partly due to changes in the responsiveness to catecholamines. Previous experiments have basically been concerned with uterine adrenoceptor binding characteristics. In the present study we have evaluated total myometrial DNA, beta 2-adrenoceptor mRNA and isoproterenol-induced relaxation of rat isolated uterine strips pre contracted with potassium on days 0, 7, 14 and 21 of pregnancy and on day 5 post partum. Total myometrial DNA expressed per milligram wet tissue peaked at day 14 of pregnancy followed by a decrease at the end of gestation. This suggests that hyperplasia predominates in the growth of the uterus in early gestation, whereas hypertrophy may be more marked in late pregnancy. The concentration of beta 2 adrenoceptor mRNA decreased linearly throughout the gestational period (0.73 +/- 0.20 amol/mg wet tissue on day 0 vs 0.34 +/- 0.09 amol/mg wet tissue on day 21, P < 0.05). Five days after parturition, at which time the uterus had returned to its pre-pregnant weight, beta 2-adrenoceptor mRNA was found to have increased 8 fold (2.79 +/- 0.14 amol/mg wet tissue, P < 0.05) as compared with day 21. The maximal effect of isoproterenol on pre-contracted uterine strips in which alpha receptors were blocked by phentolamine showed a similar decrease which on day 21 reached 67% of the day 0 level (P < 0.001). EC50 values were unchanged in all groups except day 21 pregnant rats in which an increase was observed. One-way ANOVA with Bonferroni's t-test showed statistically significant differences only between the day 21 group and either the day 5 post-partum group or the day 14 pregnant group (P < 0.05). The observed alteration in EC50 prior to the end of gestation indicates that the system becomes less sensitive to beta 2-adrenergic stimulation at this time. We conclude that a reduction of de novo synthesis of beta 2-adrenoceptors may play a role in contributing to the increased myometrial activity at term. We further suggest that the dramatic up-regulation of beta 2 adrenoceptor mRNA postpartum may protect the fully involuted uterus against excessive contractions induced by oxytocin secreted during lactation. PMID- 9203996 TI - Effects of hyperprolactinaemia on glucose tolerance and insulin release in male and female rats. AB - It has been shown that prolactin (PRL) induces glucose intolerance, hyperinsulinaemia and insulin resistance in several animal species, including rats. However, the sex differences regarding glucose homeostasis and insulin release in hyperprolactinaemic subjects have not been assessed to date. In the present study, hyperprolactinaemic (pituitary-grafted) or control (sham-operated) male and female rats were submitted to an i.v. glucose tolerance test (30 mg/100 g body weight, 30% glucose). Grafted female rats had fasting plasma glucose concentrations 26% above control (P < 0.01). After the glucose load there was a rapid and pronounced increase in plasma glucose levels in all animal groups, followed by a return to basal values within 30 min. However, the glucose concentrations in hyperprolactinaemic rats were significantly greater than those in controls at 5 min (males, P < 0.05) and 30 min (females, P < 0.05). The glucose disappearance rate was significantly increased in the grafted females compared with control (P < 0.01) and slightly increased in the grafted males. Plasma insulin concentration increased just after glucose load and returned to basal values within 5 min in all groups except for the grafted females, which had recovered their basal insulin levels at 15 min. The grafted male rats had insulin concentrations higher than those of sham-operated controls at 2 min (28.9 +/- 3.6 vs 17.3 +/- 2.1 microU/ml, P < 0.01), whereas females had plasma insulin concentrations greater than those in sham-operated controls 10 min after the glucose load (15.9 +/- 1.9 vs 10.1 +/- 1.4 microU/ml, P < 0.05). The areas under the plasma insulin concentration-time curves were also significantly increased in the hyperprolactinaemic rats and were positively correlated with plasma PRL concentrations (r = 0.613, P < 0.01). These results demonstrate that moderate chronic hyperprolactinaemia is associated with increased glucose-induced insulin release, which was altered at different times after the glucose load in grafted male and female rats, whereas fasting hyperglycaemia was observed only in grafted females, indicating a sexual dimorphism in the diabetogenic effects of PRL in rats. PMID- 9203994 TI - Effect of insulin-induced hypoglycaemia on secretion patterns and rates of corticotrophin-releasing hormone, arginine vasopressin and adrenocorticotrophin in horses. AB - To study the effect of hypoglycaemia on secretion rates of corticotrophin releasing hormone (CRH), arginine vasopressin (AVP) and ACTH in a non-ruminant species, a non-surgical method was used to collect pituitary venous (PitVen) blood every 0.5 or 1 min from seven horses before and after insulin administration (0.4 U/kg i.v.). To assess the effect of PitVen cannulation on results, peripheral hormones were also measured before and after insulin in five horses without PitVen cannulae. Insulin administration lowered plasma glucose in all horses (P < 0.0001; paired t-test). Cortisol concentrations, which were similar in horses with and without PitVen cannulae before insulin, rose significantly after insulin administration in both groups. Most horses showed discomfort as glucose fell. When data from horses with and without PitVen cannulae were pooled, the peak fractional change in cortisol (Spearman's rank correlation coefficient (rs) = -0.94, P < 0.001) and the severity of hypoglycaemic symptoms (rs = -0.61, P < 0.02) were inversely ranked with the glucose nadir. In horses with PitVen cannulae, insulin administration increased secretion rates of ACTH (P < 0.0001), AVP (P < 0.0001) and CRH (P < 0.02). Increments in ACTH (rs = -0.96, P < 0.005) and CRH (rs = -0.81, P < 0.05), but not in AVP, measured during the second half-hour after insulin (i.e. the peak response), were inversely ranked with the glucose nadir. Moreover, ACTH increments were positively ranked with those in CRH (rs = 0.81, P < 0.05), but not in AVP. Nevertheless, in individual horses, minute-to-minute AVP and ACTH concentrations in PitVen blood were always correlated, whereas minute-to-minute CRH and ACTH concentrations were correlated only when glucose dropped below 3.4 mmol/l. In less hypoglycaemic horses, ACTH secretion rose despite little or no change in CRH. We suggest that in horses AVP is the primary acute signal for ACTH release both before and during hypoglycaemia; however, the increasing magnitude of ACTH increments induced by greater degrees of hypoglycaemia is determined largely by selective CRH release, which then augments corticotroph responses to AVP. PMID- 9203995 TI - Urinary excretion of the TRH-like peptide pyroglutamyl-glutamyl-prolineamide in rats. AB - TRH-like immunoreactivity (TRH-LI) was estimated in methanolic extracts of rat tissues and blood by RIA using antiserum 4319, which binds most peptides with the structure pGlu-X-ProNH2, or antiserum 8880, which is specific for TRH (pGlu-His ProNH2). TRH-LI (determined with antiserum 4319) and TRH (determined with antiserum 8880) contents were 8 and 8 ng/g in brain, 216 and 222 ng/g in hypothalamus, 6.5 and 6 ng/g in pancreas, 163 and 116 ng/g in male pituitary, 105 and 77 ng/g in female pituitary, 1 and 0.1 ng/g in salivary gland, 61 and 42 ng/g in thyroid, 12 and 3 ng/g in adrenal, 3 and 0.3 ng/g in prostate, and 11 and 0.8 ng/g in ovary respectively. Blood TRH-LI (antiserum 4319) and TRH (antiserum 8880) levels were 31 and 18 pg/ml in male rats, and 23 and 10 pg/ml in female rats respectively. Unextracted serum obtained from blood kept for at least 1 h at room temperature no longer contained authentic TRH but still contained TRH-LI (males 20.3 +/- 3.1, females 15.9 +/- 3.0 pg/ml; means +/- S.E.M.). Isocratic reverse-phase HPLC showed that TRH-LI in serum is largely pGlu-Glu-ProNH2 (< EEP NH2), a peptide previously found in prostate and anterior pituitary. In urine, TRH-LI (antiserum 4319) and TRH (antiserum 8880) levels were 3.21 +/- 0.35 and 0.32 +/- 0.04 ng/ml in male rats and 3.75 +/- 0.22 and 0.37 +/- 0.04 ng/ml in female rats respectively (means +/- S.E.M.). Anion-exchange chromatography on QAE Sephadex showed that urine of normally fed rats contains both basic/neutral TRH LI (b/n TRH-LI) and acidic TRH-LI (aTRH-LI) in a ratio of approximately 40:60, and further analysis by HPLC indicated that aTRH-LI represents < EEP-NH2. Analysis of food extracts and urine from fasted rats demonstrated that b/n TRH-LI is derived from food particles spilled by the rats during urine collection, while aTRH-LI is endogenously produced. While urinary aTRH-LI levels were higher in female than in male rats (2.99 +/- 0.41 vs 2.04 +/- 0.20 ng/ml), the daily urinary excretion was similar in both sexes (females 15.6 +/- 1.4, males 19.5 +/- 2.0 ng/day). Intravenously injected < EEP-NH2 disappeared from serum with a half life of approximately 1 h, and was recovered unchanged and quantitatively in urine. In contrast, when < EEP-NH2 was administered with food, only approximately 0.5% was recovered in urine. The urinary clearance rate of serum TRH-LI amounted to 0.52 +/- 0.10 ml/min in males and 0.34 +/- 0.05 ml/min in females. In view of the presence of < EEP-NH2 in the anterior pituitary gland, and the regulation of its content in parallel with gonadotrophins, we examined the possibility that serum < EEP-NH2 is of pituitary origin and correlates with gonadotrophin secretion. However, treatments that alter pituitary < EEP-NH2 content and gonadotrophin release had no effect on serum TRH-LI or urinary aTRH-LI. In conclusion, the TRH-like peptide < EEP-NH2 is present in rat serum and is excreted into the urine. Moreover, < EEP-NH2 in serum and urine is not derived from rat food and is probably not of pituitary origin. PMID- 9203997 TI - Involvement of bombesin in spermine-induced corticosterone secretion and intestinal maturation in suckling rats. AB - In this study we investigated whether brain-gut peptides are implicated in the activation of the hypophysial-adrenal axis (HAA) in suckling rats treated orally with spermine. The first group of rats received i.p. injections of bombesin, vasoactive intestinal polypeptide (VIP), somatostatin or neurotensin, starting on day 11 of life, and killed on day 14. The small intestine was removed and analysed for its content of proteins, DNA, polyamines and for its specific activity (SA) of disaccharidases. The second group of rats received one of the hormones cited above and was killed 45 min after the treatment for determination of corticosterone plasma concentration. Rats of the third group were adrenalectomised then treated with bombesin as the first group. The fourth group of rats was orally treated with spermine and sacrificed 2, 3, 4, 6 and 8 h thereafter for analysis of plasma and intestinal concentrations of bombesin. The i.p. injection of bombesin increased the sucrase and maltase SA in the whole small intestine, while it decreased the lactase SA in the distal part. Intestinal weight and length, contents of DNA, protein, spermidine and spermine, and corticosterone plasma levels were enhanced by bombesin treatment. Somatostatin, neurotensin and VIP were ineffective on all the parameters studied. Adrenalectomy, in bombesin-treated rats, decreased the sucrase and maltase SA in the whole intestine, and decreased the lactase SA in the proximal intestine. It has no effect on intestinal weight and length, and protein content. Oral administration of spermine had no effect on plasma concentration of bombesin, whereas it decreased the content of this peptide in the whole small intestine. It is possible that bombesin may control intestinal development in suckling rats and be a link between the ingestion of spermine and the liberation of corticosterone by the adrenal glands. PMID- 9203998 TI - Ca2+ receptor mRNA and protein increase in the rat parathyroid gland with advancing age. AB - Parathyroid hormone (PTH) release is regulated by extracellular calcium through a Ca2+ receptor (CaR) located on the surface of the parathyroid cell. With advancing age, the serum concentration of PTH increases, and evidence suggests that the calcium set-point for PTH release may also increase. To determine whether these changes are linked to a change in CaR expression, we quantitated mRNA and protein for the receptor in parathyroid glands of 6-week-, 6-month- and 24-month-old rats. Thyroid and kidney tissue were also studied. Between 6 weeks and 24 months of age, CaR mRNA in the parathyroid gland increased 11.4- and 3.3 fold as measured by competitive reverse transcription PCR and solution hybridization assays respectively. Message levels for the receptor also increased in the thyroid but not in the kidney. Coincident with the increase in message levels, receptor protein concentration in the parathyroid increased 7-fold between 6 weeks and 24 months of age. These results suggest that the altered relationship between extracellular calcium and PTH release observed in aging is associated with dramatic changes in CaR metabolism. That PTH secretion is increased despite increased receptor concentration suggests that aging may impair calcium binding or coupling between the CaR and down-stream effector elements in the pathway regulating PTH release. PMID- 9203999 TI - Measurement of N- and C-terminal-region fragments of parathyroid hormone-related peptide in milk from lactating women and investigation of the relationship of their concentrations to calcium in milk. AB - Parathyroid hormone-related peptide (PTHrP) is found in very high concentrations in the milk of various mammals. However, little is known about its physiological role in this fluid. To obtain detailed profiles of PTHrP in milk, we measured the concentrations of PTHrP in human milk by two different region-specific assays, PTHrP(1-87) (N-PTHrP) and PTHrP(109-141) (C-PTHrP). We also examined the correlations between PTHrP and Ca concentrations in milk as well as the correlations between PTHrP and secreted milk volume. The levels of N-PTHrP and C PTHrP were relatively low after delivery and gradually increased to 13.87 +/- 2.40 nmol/l (mean +/- S.E.M.) and 56.39 +/- 11.31 nmol/l respectively on the 10th day postpartum. N-PTHrP concentration remained steady until the 6th month postpartum when weaning starts, at which point it decreased slightly. C-PTHrP levels changed in a similar way to N-PTHrP levels but were 2- to 5-fold higher. Milk Ca concentration, and content, correlated with C-PTHrP concentration, and content (r = 0.422 and r = 0.769 respectively; P < 0.0001) but not with N-PTHrP. N-PTHrP concentration in the milk samples on the 4th day postpartum correlated with the volume of milk secreted during the 24 h before the samples were taken (r = 0.524, P < 0.01), but C-PTHrP concentration did not. These results suggest that PTHrP in human milk may play some role in the maintenance of lactation through the N-terminal region and in promoting Ca transfer into milk via the C-terminal region. PMID- 9204000 TI - Intracellular regulation of 17 beta-hydroxysteroid dehydrogenase type 2 catalytic activity in A431 cells. AB - There is growing evidence that various isoforms of 17 beta-hydroxysteroid dehydrogenase (17-HSD) are regulated at the level of catalysis in intact cells. A number of investigators have proposed that the NAD(P)/NAD(P)H ratio may control the direction of reaction. In a previous study, we obtained evidence that A431 cells, derived from an epidermoid carcinoma of the vulva, are enriched in 17-HSD type 2, a membrane-bound isoform reactive with C18 and C19 17 beta hydroxysteroids and 17-ketosteroids. The present investigation was undertaken to confirm the presence of 17-HSD type 2 in A431 cells and to assess intracellular regulation of 17-HSD at the level of catalysis by comparing the activity of homogenates and microsomes with that of cell monolayers. Northern blot analysis confirmed the presence of 17-HSD type 2 mRNA. Exposure of cells to epidermal growth factor resulted in an increase in type 2 mRNA and, for microsomes, increases in maximum velocity (Vmax) with no change in Michaelis constant (Km) for testosterone and androstenedione, resulting in equivalent increases in the Vmax/Km ratio consistent with the presence of a single enzyme. Initial velocity data and inhibition patterns were consistent with a highly ordered reaction sequence in vitro in which testosterone and androstenedione bind only to either an enzyme-NAD or an enzyme-NADH complex respectively. Microsomal dehydrogenase activity with testosterone was 2- to 3-fold higher than reductase activity with androstenedione. In contrast, although cell monolayers rapidly converted testosterone to androstenedione, reductase activity with androstenedione or dehydroepiandrosterone (DHEA) was barely detectable. lactate but not glucose, pyruvate or isocitrate stimulated the conversion of androstenedione to testosterone by monolayers, suggesting that cytoplasmic NADH may be the cofactor for 17-HSD type 2 reductase activity with androstenedione. However, exposure to lactate did not result in a significant change in the NAD/NADH ratio of cell monolayers. It appears that within A431 cells 17-HSD type 2 is regulated at the level of catalysis to function almost exclusively as a dehydrogenase. These findings give further support to the concept that 17-HSD type 2 functions in vivo principally as a dehydrogenase and that its role as a reductase in testosterone formation by either the delta 4 or delta 5 pathway is limited. PMID- 9204001 TI - Transcriptional and post-transcriptional regulation of FSH receptor in rat granulosa cells by cyclic AMP and activin. AB - The effect of FSH on the induction of FSH receptors in granulosa cells is believed to be mediated, at least in part, by the cAMP second messenger system. We examined the effect of activin and cAMP on FSH receptor expression in this culture system. Steady-state levels of FSH receptor mRNA, analyzed by Northern blot hybridization, increased 3.5-fold in response to 24-h incubation with activin and 1.7-fold with 12-h incubation with 8-bromoadenosine 3,5-cyclic monophosphate (8-Br-cAMP; 0.2 mM). We have investigated whether 8-Br-cAMP- and/or activin-induced increases in FSH receptor mRNA levels are the result of increased transcription and/or altered mRNA stability. The rates of FSH receptor mRNA gene transcription, assessed by nuclear run-on transcription assay, increased 3-fold in cells treated with activin and 1.5-fold in cells treated with 8-Br-cAMP for 2 h. To examine the degradation rates of FSH receptor mRNA transcripts, granulosa cells were preincubated with 8-Br-cAMP, activin, or medium alone for 6 h. After the preincubation period, 5 microM actinomycin-D or 200 microM 5,6-dichloro-1 beta-ribofuranosyl benzimidazole were added to arrest new RNA synthesis. The decay curves for the 2.4 kb FSH receptor mRNA transcript in granulosa cells were not significantly different in the absence or presence of 8-Br-cAMP. Activin, on the other hand, significantly altered the slope of the FSH receptor mRNA decay curve and increased the half-life of the 2.4 kb FSH receptor mRNA transcript. These data provide evidence that cAMP induces FSH receptor mRNA levels by stimulating the transcription rate and that activin increases FSH receptor mRNA levels both by stimulating transcription rates and by stabilizing the FSH receptor mRNA transcripts. PMID- 9204002 TI - Modulation of human neutrophil function in vitro by gastrin. AB - We have studied the effects in vitro of gastrin-17 and gastrin-34, at concentrations from 10(-14) M to 10(-6) M, on several of the functions of peripheral blood human neutrophils, i.e. adherence to substrate, mobility (spontaneous and directed by a chemical gradient or chemotaxis), ingestion of inert particles (latex beads) and cells (Candida albicans) and superoxide anion production. Both gastrins inhibited several steps of the phagocytic process of human neutrophils, such as mobility and ingestion. By contrast, these peptides increased adherence and had no effect on superoxide anion production. In general, these effects were significant at peptide concentrations between 10(-12) M and 10(-8) M with a maximal effect at 10(-10) M. In addition, gastrin peptides induced a significant increase in intracellular cAMP levels at 30, 60 and 120 s. Moreover, the inhibitory effect of gastrin-17 on the ingestion capacity of neutrophils (latex bead phagocytosis) was similar to that obtained with EGTA, a well-known extracellular calcium chelating compound. Gastrin-17 was found to inhibit completely the stimulation of latex bead phagocytosis in neutrophils caused by the calcium ionophore A23187. These results suggest that gastrin is a negative modulator of the phagocytic process of human neutrophils, and that this effect might involve an increase in intracellular cAMP levels and a decrease in calcium entry into the cells. PMID- 9204003 TI - Immunolocalisation of oestrogen receptor-alpha within the testis and excurrent ducts of the rat and marmoset monkey from perinatal life to adulthood. AB - The sites of action and the physiological role of oestrogens in the male reproductive tract are poorly understood. We have undertaken a systematic study of the immunoexpression of oestrogen receptor-alpha (ER alpha) in the male rat from late fetal life through to adulthood and compared the findings with results obtained in the marmoset monkey (Callithrix jacchus) from neonatal to adult life. The testes, rete testis, efferent ducts and epididymis were examined from normal male rats (aged 4, 8, 10, 15, 20, 25, 38, 48 and 90 days) and from male rat fetuses on days 17.5 and 18.5 of gestation; comparable tissues were examined from neonatal, infantile, peripubertal and adult marmosets aged 8, 18-24, 54-62 and 92 112 weeks respectively. Immunolocalisation of ER alpha used antigen retrieval and a monoclonal antibody directed to the N-terminus, which had proved superior to six other antisera tested. ER alpha was immunoexpressed in interstitial cells, including the fetal/ neonatal generation of Leydig cells, in both the rat and marmoset. In the rat, the adult generation of Leydig cells were also immunopositive for ER alpha whereas the comparable cells in the marmoset were only weakly immunopositive. ER alpha was not expressed in Sertoli cells, peritubular myoid cells, blood vessels or germ cells at any time in either species. In late fetal life in the rat, ER alpha was immunoexpressed in cells surrounding the mesonephric tubules, whereas postnatally it was expressed in the epithelium of the rete testis and efferent ducts at all ages from 4 to 90 days; this immunoexpression was most pronounced in the efferent ducts. In the marmoset, the efferent ducts, but not the rete testis, also showed intense immunoexpression of ER alpha. Apart from sporadic immunostaining for ER alpha in the epididymal duct of the rat in the neonatal period, the caput, corpus and cauda epididymis were negative for immunoexpression of ER alpha at all ages in both species. These findings suggest that the main actions of oestrogens in the male reproductive tract, mediated by ER alpha, are related to the development and function of the efferent ducts and the Leydig cells. In consideration of data from this and previous studies of oestrogen binding, we predict possible sites of expression of other oestrogen receptors (e.g. ER beta) in Sertoli cells and the epididymis. Interactive effects, related to the relative levels of androgens and oestrogens, could be physiologically important in the excurrent ducts of the adult testis. PMID- 9204004 TI - New treatments in adult cystic fibrosis. PMID- 9204005 TI - Ursodeoxycholic acid in cystic fibrosis-related liver disease: a systematic review. PMID- 9204007 TI - Cystic fibrosis nurse specialist: a key role. PMID- 9204006 TI - Joint disorders in cystic fibrosis. PMID- 9204008 TI - Home intravenous antibiotic therapy: practical aspects in children. PMID- 9204009 TI - Home intravenous antibiotic therapy: practical aspects in adults. PMID- 9204010 TI - Quality of life in cystic fibrosis. PMID- 9204011 TI - Gene therapy: the case for cystic fibrosis. PMID- 9204012 TI - Management of cystic fibrosis before and after lung transplantation. PMID- 9204014 TI - Quality in general practice. PMID- 9204015 TI - Are you a commander or a guide? PMID- 9204013 TI - Subdural empyema due to Burkholderia cepacia: an unusual complication after lung transplantation for cystic fibrosis. PMID- 9204017 TI - Risk factors for ischaemic heart disease in patients with dermatitis herpetiformis. AB - For reasons that are unclear, patients with dermatitis herpetiformis (DH) have a lower than expected mortality rate from ischaemic heart disease. We have compared risk factors for ischaemic heart disease (lipids, fibrinogen levels, smoking history and social class) in 29 DH patients and 57 controls matched for age and sex. Patients with DH had significantly lower cholesterol, triglycerides, apolipoprotein B and fibrinogen and higher HDL2; they also smoked less and were of higher social class. The mechanisms underlying these observations merit further investigation. Intestinal abnormalities or gluten-free diet may account for differences in lipid fractions, and the immunomodulatory properties of cigarette smoke may protect against the development of DH. PMID- 9204016 TI - The undergraduate surgical curriculum in a changing National Health Service. PMID- 9204018 TI - Chronic fatigue syndrome: sufferers' evaluation of medical support. AB - In response to reports of negative cooperation between sufferers of chronic fatigue syndrome (CFS) and their doctors, semi-structured interviews were conducted with sufferers from two different patient samples. Satisfaction with support received and with medical professionals in general was low. Sufferers complained about insufficient informational as well as emotional support from their doctors, and as a consequence most opted for alternative or complementary forms of treatment. In addition, disagreements over illness aetiology and treatment precluded effective cooperation. If satisfaction and compliance are to improve, sufferers will need more information about CFS and more support. PMID- 9204019 TI - Occipital seizures imitating migraine aura. AB - Three cases are reported in which symptoms of occipital seizures resembled the visual aura of migraine. Careful recording of the characteristics and timing of such visual effects will often resolve the diagnostic dilemma. PMID- 9204020 TI - First two years of a follow-up breast clinic led by a nurse practitioner. AB - After special training a nurse practitioner ran an independent clinic for follow up patients with breast disease. All patients referred to a single surgical firm with breast cancer and most patients with benign disease who required follow-up were included. In the first 2 years of the service 382 clinic visits were recorded (median 5/clinic, range 1-12). The nurse practitioner reviewed 236 (62%) patients alone but involved the consultant surgeon in the remainder. No significant lesion was missed in these patients. The nurse-led clinic is popular with patients and, subject to careful supervision, offers an attractive option for follow-up of patients with breast disease. PMID- 9204021 TI - Glycobiology and medicine: an introduction. PMID- 9204023 TI - Team-working in primary care. PMID- 9204022 TI - Asthma mortality: the worldwide response. PMID- 9204024 TI - Recurrent epistaxes in hereditary haemorrhagic telangiectasia. PMID- 9204025 TI - Non-Hodgkin lymphoma with panhypopituitarism, hyperprolactinaemia and sixth nerve palsy. PMID- 9204026 TI - Strangulated femoral hernia presenting as haematemesis. PMID- 9204027 TI - Limb gangrene and two types of vasculitis. PMID- 9204028 TI - Skip lesions in pneumatosis coli. PMID- 9204029 TI - Evidence-based medicine: old French wine with a new Canadian label? PMID- 9204030 TI - A cure for the ague: the contribution of Robert Talbor (1642-81) PMID- 9204031 TI - Franz Schubert's last illness. PMID- 9204032 TI - Medical scientism. PMID- 9204033 TI - Group B streptococcal vulvovaginitis. PMID- 9204034 TI - Lind, Scott, Amundsen and scurvy. PMID- 9204035 TI - Idiopathic rhabdomyolysis. PMID- 9204036 TI - Complementary medicine in the medical curriculum. PMID- 9204037 TI - Complementary medicine in the medical curriculum. PMID- 9204038 TI - Proposed 1998 federal budget abandons nurse education. PMID- 9204039 TI - The Nursing Work Force Beyond 2000 Project. The Greater Coastal Bend Region of Texas. PMID- 9204041 TI - Strategic partnering: clinical and risk management concerns. PMID- 9204042 TI - Federal health policy directions. PMID- 9204043 TI - Critical pathways: effectiveness in achieving patient outcomes. AB - Refining the clinical care process to produce high-quality patient outcomes is becoming increasingly important as health care administrators strive for success in a mature managed care environment. This study examines the effect of structuring interventions and the evaluation of patient response, inherent in the critical pathway process, on clinical, length-of-hospital-stay, and financial patient outcomes. This study differs from previous critical pathway trials in that an objective measure of quality was used and the critical pathways were not introduced concurrently with a case management delivery model. The results show that critical pathways may be a significant determinant of improved quality in a managed care environment. The findings also suggest ways to improve nursing practice, nursing education, and nursing informatics. PMID- 9204044 TI - Survey and critique of studies related to unlicensed assistive personnel from 1975 to 1997, Part 1. AB - This descriptive integrated review of research on the use of unlicensed assistive personnel in nursing is presented in two parts. In this issue, part 1 describes the methods used to find and critique research related to unlicensed assistive personnel in nursing. It includes the conceptual model and findings related to the variables studied. Part 2 of this review, which is scheduled for publication in the next issue, will present research findings, conclusions, and recommendations. PMID- 9204045 TI - Breaking through the barriers: healthcare for the homeless. AB - National welfare reform is predicted to increase the number of homeless persons. This will affect the health care system by increasing the number of uninsured people and by multiplying the number of homeless persons seeking care in hospital emergency departments. Homeless persons have four major barriers to care: financial, bureaucratic, programmatic, and personal. The authors provide an overview of the homeless population, outline the barriers to health care for persons who are homeless, and highlight the major health care needs of this population. Finally, a community-based service delivery system developed by one agency in responding to the need of homeless persons is provided as a model of care. PMID- 9204046 TI - Creating a healthy work environment in the midst of organizational change and transition. AB - In the midst of organizational change and transition, the need for a healthy work environment is greater than ever. Leaders may be in a position of leading staff on a journey they would rather not be on. Although there may not be a choice of destination, there are many decisions to be made along the way that will impact the health and quality of the journey. Creation of a healthy work environment does not occur overnight. It requires acknowledgment of the reality of the present environment, clear behavioral expectations and standards, systems, and structures to ensure the organizational changes are enduring and a means to assess continually the health of the work environment. Leaders have an opportunity and a responsibility to structure organizations in such a way that dignity, integrity, honesty, and compassion are preserved. PMID- 9204047 TI - The effect of workplace empowerment on staff nurses' occupational mental health and work effectiveness. AB - Occupational mental health has been linked to productivity and other desired organizational outcomes, such as commitment and satisfaction. Kanter's model of work empowerment was used to examine the relation between 62 staff nurses' perceptions of empowerment in their work settings and their occupational mental health. The authors discuss their findings and suggest organizational interventions that can be used by nurse administrators to ameliorate work stress and improve work effectiveness. PMID- 9204048 TI - Identifying nursing research priorities in a newly merged healthcare system. AB - Merging of nursing services led to key opportunities to identify nursing research priorities in the authors' institution. The literature suggested that a Delphi study would best accomplish these goals. Using a 110-member panel, a study was begun that identified two final topics that were focused on nursing administration research. Collecting these data served many purposes, including increased organizational awareness about nursing research and development of a nursing research council to facilitate future activities. PMID- 9204049 TI - Futile care. PMID- 9204050 TI - Apoptosis and cutaneous biology. AB - Apoptosis, a genetically encoded program that results in cell death, represents a fundamental biologic concept that has relevance to a wide range of dermatologic processes. This review discusses the basic biology of apoptosis and its relevance to cutaneous disease. PMID- 9204051 TI - Cutaneous manifestations of chronic granulomatous disease. A report of four cases and review of the literature. AB - BACKGROUND: Chronic granulomatous disease represents a group of genetic disorders in which impaired intracellular microbial killing by phagocytes leads to recurrent bacterial and fungal infections and granuloma formation. Cutaneous disease occurs in 60% to 70% of cases. The characteristic histologic finding of pigmented lipid macrophages in visceral granulomas has not been described previously in the skin. OBJECTIVE: Our purpose was to review our experience of skin disorders in chronic granulomatous disease. METHODS: We studied the clinical and histologic findings in four patients with chronic granulomatous disease and unusual skin lesions. We reviewed the skin disorders seen in five additional patients with chronic granulomatous disease referred to the pediatric dermatology clinic. The literature was reviewed for previously reported cutaneous manifestations of chronic granulomatous disease. RESULTS: A teenage boy with chronic granulomatous colitis had nonulcerating cutaneous granulomas from which no organisms were isolated. Histologic examination of both skin and bowel revealed the characteristic golden-yellow granular pigment in macrophages. A second boy had cutaneous aspergillosis involving the left foot; histologic examination revealed macrophages containing yellow-brown pigment at the periphery of the granulomatous inflammation. Two children had vesicular skin lesions. These lesions were recurrent in one boy for several years. In the second child they were associated with fatal intracranial and pulmonary infection. Histologic examination in both cases revealed a subcorneal polymorphonuclear infiltrate and perivascular macrophages containing yellow-brown pigment. Cultures were either negative or revealed organisms that are normally nonpathogenic skin commensals, such as coagulase-negative staphylococci. CONCLUSION: The cutaneous manifestations of chronic granulomatous disease encompass a variety of infections and inflammatory lesions. Diagnostic and therapeutic problems may arise because of difficulty in isolating a causative organism. The characteristic pigmented macrophages of visceral granulomas can also be found in skin lesions. PMID- 9204052 TI - Nail lichen striatus: clinical features and long-term follow-up of five patients. AB - BACKGROUND: Nail involvement in lichen striatus (LS) is uncommon and has always been reported in association with typical skin lesions. OBJECTIVE: We attempted to characterize the clinical and pathologic features and the long-term prognosis of nail LS. METHODS: Five cases of LS of the nail including three cases with exclusive nail involvement were evaluated and the literature reviewed. RESULTS: Biopsy specimens showed a moderately dense bandlike lymphohistiocytic infiltrate affecting the proximal nailfold, the nail bed, and the nail matrix dermis. Exocytosis with slight spongiosis, focal hypergranulosis, and dyskeratotic cells were detectable in the nail matrix epithelium. Spontaneous regression of the onychodystrophy occurred after 4 to 12 months from the time of diagnosis (mean, 8.4 months). CONCLUSION: Nail LS is not necessarily associated with skin lesions but can also be an isolated finding. The diagnosis of nail LS should be strongly suspected when a child or a young patient presents with lichen planus-like nail abnormalities localized to the lateral or medial portion of a single nail. PMID- 9204053 TI - Lentigo maligna and lentigo maligna melanoma. PMID- 9204054 TI - Utility of a standard allergen series alone in the evaluation of allergic contact dermatitis: a retrospective study of 732 patients. AB - BACKGROUND: Patch testing remains the standard for the diagnosis of allergic contact dermatitis. The validity and usefulness of a standard patch test allergen series has not been addressed adequately by previous studies. OBJECTIVE: We sought to examine the utility of the standard allergen series as a sole screening tool in the diagnosis of allergic contact dermatitis. METHODS: The charts of 732 patients referred for patch testing were reviewed for positive patch test results. The group of patients with positive reactions was stratified into two groups based on the clinical relevance of their reactions. These groups were subsequently analyzed to determine whether the reactions were to part of the standard series of allergens or to part of a supplementary group. RESULTS: Of patients tested, 50% had a positive patch test. Of those, 221 (30%) had reactions deemed clinically relevant. Only 23% of patients with positive patch tests reacted to an allergen(s) in the standard series exclusively. When adjusted for clinical relevance, only 15.7% of patients were completely evaluated with the standard series of 20 allergens. CONCLUSION: The standard allergen series of 20 allergens available in the United States is limited as a screening tool when used alone in the evaluation of patients with allergic contact dermatitis. PMID- 9204055 TI - Recognition of pemphigus antigens in drug-induced pemphigus vulgaris and pemphigus foliaceus. AB - BACKGROUND: The clinical appearance and biologic behavior of drug-induced pemphigus depend on the type of inducing drug. OBJECTIVE: Our purpose was to investigate patients with drug-induced pemphigus vulgaris and pemphigus foliaceus antigens and compare results of studies to detect antibody reactivity in sera of these patients with the serology of patients with idiopathic pemphigus. METHODS: Ten patients with drug-induced pemphigus were studied. Antibody reactivity was determined against the pemphigus vulgaris antigen, desmoglein 3, and against desmoglein 1. RESULTS: The patient with pemphigus foliaceus and low levels of autoantibodies precipitated neither antigen. One patient with pemphigus vulgaris and high levels of antibody also failed to precipitate any specific antigen. Sera from eight patients with drug-induced pemphigus vulgaris had circulating autoantibodies directed to either the pemphigus vulgaris or pemphigus foliaceus antigen. Low levels of antibody in two of these eight patients precipitated only the pemphigus foliaceus antigen. High levels of antibody in five of the eight patients precipitated the pemphigus vulgaris antigen; two of these also reacted with the pemphigus foliaceus antigen. CONCLUSION: The autoantibody response was similar in both spontaneous and drug-related disease. A similar molecular mechanism in the two types of pemphigus is suggested. PMID- 9204056 TI - p53 tumor suppressor gene protein expression in premalignant and malignant skin lesions of kidney transplant recipients. AB - BACKGROUND: Kidney transplant recipients have an increased incidence of skin cancer, the cause of which is likely multifactorial. Inactivation of the tumor suppressor gene p53 protein may be important. Chemoprophylaxis of skin cancer with retinoids is beneficial in these patients. OBJECTIVE: Our purpose was to investigate the immunohistochemical expression of p53 protein in premalignant and malignant cutaneous lesions in kidney transplant recipients and the effect of low dose etretinate on p53 expression. METHODS: Paraffin sections were stained with the monoclonal antibody DO-7. RESULTS: Immunoreactivity of p53 was observed in 59% of basal cell carcinomas and more than 60% of squamous cell carcinomas, Bowen's disease, dysplastic lesions, and viral warts. No demonstrable effect of etretinate on p53 expression could be determined. CONCLUSION: The high prevalence of p53 immunoreactivity in premalignant and malignant skin lesions of kidney transplant recipients supports a role for p53 protein in skin cancer. This could be caused by mutation of the p53 gene, inactivation, or failure of degradation of p53 protein. PMID- 9204057 TI - Ranitidine does not affect psoriasis: a multicenter, double-blind, placebo controlled study. AB - BACKGROUND: Data from open studies suggest that ranitidine has a beneficial effect on psoriasis and is well tolerated. OBJECTIVE: Our purpose was to determine the effectiveness of ranitidine in a 24-week, multicenter, double blind, placebo-controlled, dose-comparing study of 201 patients with psoriasis. METHODS: Patients with moderate to severe psoriasis who had stopped systemic antipsoriatic therapy, including PUVA and UVB, for at least 10 weeks were included. After a washout period of 2 weeks, patients were randomly allocated to use either ranitidine, 150 mg twice a day; ranitidine, 300 mg twice a day; or placebo for up to 24 weeks. Assessment with the Psoriasis Area and Severity Index was performed at weeks 3, 6, 9, 12, 18, and 24 after randomization. Reduction of the Psoriasis Area and Severity Index score by 70% at the completion of the study was considered a treatment success. RESULTS: The success rates at week 24 in the 300 mg, 600 mg, and placebo groups were 11%, 5%, and 12%, respectively. No significant differences were observed between the three treatment groups at any stage of the study. CONCLUSION: This study provides strong evidence that ranitidine does not affect the skin disease in patients with psoriasis. PMID- 9204058 TI - Further evidence of the role of HLA-DR4 in the genetic susceptibility to actinic prurigo. AB - BACKGROUND: Actinic prurigo (AP) is triggered by sun exposure. Its prevalence in Mexicans seems to be particularly high, which suggests a genetic susceptibility. OBJECTIVE: Our purpose was to determine the role of major histocompatibility complex (MHC) genes in the genetic susceptibility to AP. METHODS: Fifty-six Mexican Mestizo patients with AP underwent serologic typing for HLA class I and class II antigens. Class II MHC genes were also studied by DNA analysis. Findings in patients were compared with 100 ethnically matched healthy controls. RESULTS: We found that 92.8% of patients with AP were HLA-DR4 positive (corrected p = 0.002; odds ratio [OR] = 10.1). The class I antigens HLA-A28 and HLA-B39 (B16) were also significantly increased (p < or = 0.000001, OR = 20.9 and p = 0.0001, OR = 6.7, respectively) compared with normal controls. Allele-specific oligonucleotide DR4 subtyping showed that 80.7% of HLA-DR4+ patients with AP were also positive for the DRB1*0407 allele. CONCLUSION: These results confirm the role of HLA-DR4 (DRB1*0407) in the genetic susceptibility to AP and raise the possibility of a role for class I MHC antigens HLA-A28 and B16 in Mexican patients. PMID- 9204059 TI - High-dose UVA1 radiation therapy for localized scleroderma. AB - BACKGROUND: Fibrotic skin lesions in patients with localized scleroderma can cause muscle atrophy, disfigurement, and flexion contractures. There is no effective therapy for this disease. Skin fibrosis is thought to be caused by decreased collagenase activity. Collagenase activity can be induced in dermal fibroblasts by UVA1 irradiation. OBJECTIVE: Our purpose was to assess whether UVA1 radiation therapy is effective for patients with localized scleroderma. METHODS: Patients with localized scleroderma (n = 17) were exposed 30 times to 130 J/cm2 UVA1 (high-dose UVA1 therapy; n = 10) or 20 J/cm2 UVA1 (low-dose UVA1 therapy; n = 7). Therapeutic effectiveness was assessed by evaluation of (1) clinical features, (2) thickness of sclerotic plaques, and (3) cutaneous elastometry. Sequential biopsy specimens from treated lesions were analyzed for collagenase I messenger RNA (mRNA) expression by semiquantitative reverse transcriptase-polymerase chain reaction. RESULTS: In all patients, high-dose UVA1 therapy softened sclerotic plaques, and complete clearance was observed in four of 10 patients. High-dose UVA1 therapy significantly reduced thickness and increased elasticity of plaques. These changes could not be detected in unirradiated control plaques and were still present in 9 of 10 patients 3 months after cessation of therapy. For all factors assessed, high-dose UVA1 was superior to low-dose UVA1 therapy (p = 0.001). High-dose UVA1 therapy increased collagenase I mRNA expression about 20-fold in treated plaques. CONCLUSION: High dose UVA1 therapy is effective in the treatment of localized scleroderma. Effectiveness is UVA1 dose dependent and is associated with induction of collagenase I expression. PMID- 9204060 TI - Bath-5-methoxypsoralen-UVA therapy for psoriasis. AB - BACKGROUND: After oral intake, 5-methoxypsoralen (5-MOP) is as effective as 8-MOP for PUVA therapy for psoriasis, with a lower incidence of acute cutaneous side effects. OBJECTIVE: We compared bath-water delivery of 5-MOP and 8-MOP for photochemotherapy of psoriasis. METHODS: Twenty-two patients underwent phototesting with 0.0003% 5-MOP or 8-MOP aqueous solutions. Twelve patients with palmar psoriasis were studied with a side-to-side comparison, and 10 patients with recurrent plaque-type psoriasis were treated with one therapy or the other. RESULTS: Minimal phototoxic dose (MPD) values were 2.8 +/- 1.2 J/cm2 with 8-MOP and 2.0 +/- 1.2 J/cm2 with 5-MOP (p < 0.01). Both therapies cleared palmar lesions but 8-MOP required more UVA irradiation (46.3 +/- 21.0 J/cm2 vs 30.2 +/- 21.5 J/cm2; p < 0.01) and more exposures (21.0 +/- 6.0 vs 17.0 +/- 5.0; p = 0.02). Bath-5-MOP-UVA was also more effective in the treatment of plaque-type psoriasis (cumulative UVA doses, 56.8 +/- 39.2 vs 59.1 +/- 27.9 J/cm2; number of exposures, 20.0 +/- 5.7 vs 21.6 +/- 4.7), but these differences were not significant (p = NS). Patients developed an intense tan significantly earlier with 5-MOP than with 8-MOP (3.5 +/- 0.5 weeks vs 4.4 +/- 0.5 weeks; p < 0.01). CONCLUSION: Bath-5-MOP-UVA was more phototoxic than bath-8-MOP-UVA. It was more effective in the treatment of palmar psoriasis, whereas its greater pigmentogenic activity appeared to have an adverse effect on therapeutic effectiveness in the treatment of plaque-type psoriasis. PMID- 9204061 TI - Pentostatin (2'-deoxycoformycin) in the treatment of cutaneous T-cell lymphoma. AB - BACKGROUND: The treatment of patients with advanced or therapy-refractory cutaneous T-cell lymphoma (CTCL) remains a challenge. Pentostatin is a potent inhibitor of adenosine deaminase and is selectively toxic to lymphocytes. In a small number of patients with CTCL, it previously has been shown to be effective. OBJECTIVE: Our purpose was to evaluate the efficacy and safety of pentostatin in the treatment of patients with advanced and/or therapy-refractory CTCL. METHODS: Eighteen patients with stage I to IVb CTCL were treated with 4 to 5 mg/m2 of intravenous pentostatin every 1 to 4 weeks. RESULTS: Two patients (11%) had complete responses of 4 months and 6 years, respectively. These patients had stage III and IVa CTCL and had previously received many different external or systemic treatments. Partial remission (50% to 99% clearing) lasting for 1.5 to 6 months occurred in five patients (28%) with stage IIa (n = 3), stage IIb, and stage IVa CTCL. These patients had received a median of three prior external or systemic treatments. No major side effects were observed, and bone marrow suppression was mild. CONCLUSION: Single-agent pentostatin in intravenous doses of 4 to 5 mg/m2 is an effective systemic treatment of CTCL (39% objective response rate) with little toxicity. PMID- 9204062 TI - 5-fluorouracil iontophoretic therapy for Bowen's disease. AB - BACKGROUND: Topical 5-fluorouracil (5-FU) is an accepted therapy for Bowen's disease. Recurrences with this method have been attributed to deep follicular involvement and poor patient compliance because of the prolonged treatment time required. OBJECTIVE: We sought to determine whether iontophoresis of 5-FU is an effective therapy for Bowen's disease. METHODS: Twenty-six patients with biopsy proven Bowen's disease received eight 5-FU iontophoretic treatments in 4 weeks. Local excision was done 3 months after the last treatment. The specimens were step-sectioned and evaluated for any histologic evidence of bowenoid changes. RESULTS: Only 1 of 26 patients showed histologic evidence of Bowen's disease 3 months after treatment. CONCLUSION: 5-FU iontophoresis appears to be a safe, effective, and well-tolerated therapy for Bowen's disease. PMID- 9204063 TI - Adult pityriasis rubra pilaris: a 10-year case series. AB - BACKGROUND: Pityriasis rubra pilaris (PRP) often has a devastating impact on the lives of patients. Descriptions of its histopathologic features are not uniform. Finding a successful therapy can be challenging. OBJECTIVE: Our purpose was to examine the histopathologic features and response of patients to our standard therapy of an oral retinoid and concomitant or later addition of low-dose weekly methotrexate. METHODS: A retrospective chart review was done on 24 patients with PRP seen from March 1986 to March 1996. Biopsy specimens from 19 patients were reexamined. Telephone follow-up was conducted to determine maintenance of remission. RESULTS: All patients had the adult acquired form of PRP. Biopsy specimens from nine patients were characterized by prototypical findings of PRP, while the others included both typical and other features. Twenty-two patients were treated with either isotretinoin, 40 mg twice daily, or etretinate, 25 to 75 mg/day. Six patients with more disabling involvement had low-dose weekly methotrexate ranging from 5 to 30 mg started concurrently. Five patients had weekly methotrexate added at a later time. Seventeen patients showed 25% to 75% response after 16 weeks of therapy. All patients whose skin cleared maintained their remission. CONCLUSION: Initial oral retinoid plus concurrent or later low dose weekly methotrexate resulted in 25% to 75% improvement of PRP in 17 of 24 patients after 16 weeks of therapy. Some of the atypical features seen in biopsy specimens emphasize the importance of clinical and histopathologic correlation in establishing the diagnosis. PMID- 9204064 TI - The nasofacial interpolated flap in reconstruction of the nasal ala. AB - BACKGROUND: Skin cancer frequently involves the nasal alae, the surgical reconstruction of which may be challenging if their margins, contours, and surface texture are to be preserved. OBJECTIVE: Our purpose was to describe our experience with a nasofacial interpolated flap in which a temporary bridging pedicle is used to transpose skin from the nasofacial and melolabial sulci to an alar defect. METHODS: The nasofacial interpolated flap was used in eight patients to reconstruct partial-thickness alar wounds after excision of a basal cell carcinoma. RESULTS: The functional and cosmetic outcome was excellent in five patients and is likely to be good in two others who had postoperative fullness of the flap inset. A poor result was seen in one patient, a heavy smoker, who healed with an atrophic scar at the margin of the primary defect. CONCLUSION: In alar wounds for which a full-thickness skin graft would provide inadequate bulk, the nasofacial interpolated flap transposes skin of excellent color and textural match without blunting the nasofacial sulcus or the alar groove. PMID- 9204065 TI - Antiphospholipid syndrome and the skin. AB - The antiphospholipid syndrome is an acquired multisystem disorder of hypercoagulation, which may be primary or secondary to underlying diseases. Serologic markers for the syndrome are the lupus anticoagulant and anticardiolipin antibodies. Clinical features include recurrent thrombotic events (arterial or venous), repeated fetal loss, and thrombocytopenia. Cutaneous manifestations may occur as the first sign of antiphospholipid syndrome. These include livedo reticularis, necrotizing vasculitis, livedoid vasculitis, thrombophlebitis, cutaneous ulceration and necrosis, erythematous macules, purpura, ecchymoses, painful skin nodules, and subungual splinter hemorrhages. Antiphospholipid syndrome may also be associated rarely with anetoderma, discoid lupus erythematosus, cutaneous T-cell lymphoma, or disorders that closely resemble Sneddon or Degos syndromes. Noninflammatory vascular thrombosis is the most frequent histopathologic feature observed. Prophylaxis and treatment of thrombosis in patients with antiphospholipid syndrome relies principally on anticoagulant and antiplatelet agents. PMID- 9204066 TI - Recent developments in the treatment of atopic eczema. AB - Atopic eczema remains a therapeutic challenge. However, new developments in the understanding of the pathogenesis of this complex disease have prompted new therapeutic strategies. This review focuses on recently described treatment modalities for atopic eczema that are currently available or under investigation. The effectiveness of phototherapy, cytokines, and immunosuppressive drugs is evaluated. In addition, some new and promising but still experimental approaches are discussed. PMID- 9204067 TI - Log-in/log-out time: a quality factor for a reference laboratory--prolonged times for skin pathology processing in managed care-authorized laboratories. AB - Managed care organizations may divert skin biopsy specimens to commercial laboratories selected on a cost basis. Diversion to these laboratories could result in service of decreased quality for the patient and referring physician. Log-in/log-out dates were collected for all specimens submitted to managed care authorized laboratories either from a university-based clinic or from a private practitioner's office for a period of 18 months and compared with data obtained from a local dermatopathology laboratory. A subgroup of specimens containing inflammatory diagnoses or nondiagnostic changes was also examined. Mean log in/log-out times were 1.338 days in the dermatopathology laboratory, 6.123 days in managed care-authorized laboratories from the university site, and 7.798 days in managed care-authorized laboratories from a practitioner's office. The differences between the dermatopathology laboratory log-in/log-out times and those of the managed care-authorized laboratories were statistically significant (p < 0.0001). The conclusion from this study is that a quality indicator defined as time from log in to log out revealed a significant increase in interpretation time at managed care-designated laboratories. Although managed care plans can decrease their financial risk by contracting with national laboratories to provide all services for a set fee, a decreased quality of service can be demonstrated. PMID- 9204068 TI - Outcomes research: an overview. AB - During the past few years there has been significant interest in studying methods that document outcomes of medical care. Outcomes management should result in higher quality health care at lower cost. However, what does outcomes research mean and how does it apply to dermatology and specifically to the individual dermatologist? This article reviews the evolution of medical outcomes research and presents the status of the current instruments, indices, and methods. PMID- 9204070 TI - Surgical pearl: the self-removable suture. PMID- 9204069 TI - Surgical pearl: use of nasal packing with airway after reconstruction of deep nasal defects. PMID- 9204071 TI - Improving the cosmetic appearance of photoaged skin with glycolic acid. PMID- 9204072 TI - Iododerma after computed tomographic scan with intravenous radiopaque contrast media. PMID- 9204073 TI - Phenytoin-like hypersensitivity associated with lamotrigine. PMID- 9204075 TI - Prevalence of atopic dermatitis in patients with Down syndrome: a clinical survey. PMID- 9204074 TI - Hypersensitivity reaction to terbinafine. PMID- 9204076 TI - Trichilemmomal carcinoma in a patient with Cowden's disease (multiple hamartoma syndrome). PMID- 9204077 TI - Oral erosive lichen planus with epidermolytic hyperkeratosis during interferon alfa-2b therapy for chronic hepatitis C virus infection. PMID- 9204078 TI - Isolated cutaneous epithelioid hemangioendothelioma. PMID- 9204079 TI - Acne inversa (pyodermia fistulans sinifica) and smoking. PMID- 9204080 TI - Cyclosporine in children with severe atopic dermatitis. PMID- 9204081 TI - A flexible scalpel for shave excision of skin lesions. PMID- 9204082 TI - Suturing--control and conservation. PMID- 9204083 TI - Suturing. PMID- 9204084 TI - The treatment of acne with topical retinoids: one man's opinions. AB - Acne is a family of disorders that vary greatly in pathogenesis and clinical manifestation. Accordingly, no simple recipe for treatment can be given, and treatment options vary with the stage and intensity of the disease. Topical retinoids are the mainstay for treating common varieties of acne vulgaris. They also prevent development of comedones, halting progression to inflammatory lesions. Tretinoin was the first retinoid used in the topical treatment of acne more than 25 years ago. Isotretinoin, which has recently become available, is less irritating, but is probably somewhat less effective. Adapalene is a recently introduced topical retinoid used to treat acne. It enjoys therapeutic equivalence to tretinoin but is less irritating. Except for very mild acne cases, topical retinoids should be used concommitantly with other drugs. The operating principle is to choose drugs whose modes of action are different from topical retinoids, that is, antibiotics or benzoyl peroxide. Topical retinoids, however, constitute the core of nearly all therapeutic programs for acne. PMID- 9204085 TI - Pharmacology and chemistry of adapalene. AB - BACKGROUND: Retinoid research in the field of dermatology has been influenced by the clinical success of topical tretinoin and oral isotretinoin in the treatment of acne, and by the discovery of high-affinity binding proteins for retinoic acid mediating its action and interaction with other vitamins and hormones. OBJECTIVE: We sought molecules with an optimal balance between stability, efficacy, and local tolerance for topical acne therapy. METHODS: In vitro and in vivo bioassay systems were used to test the ability of retinoids to modulate cell proliferation and differentiation. In addition, antiinflammatory properties were assessed. Binding and transactivation assays were used to compare affinities and transcriptional activities of adapalene and tretinoin for the nuclear transcription factors, retinoic acid receptors (RARs). RESULTS AND CONCLUSION: Adapalene is a stable naphthoic acid derivative with potent retinoid pharmacology, controlling cell proliferation and differentiation. In addition it has significant antiinflammatory action. The nuclear gene transcription factors RAR beta and RAR gamma mediate the retinoid activity of adapalene. PMID- 9204086 TI - Adapalene 0.1% gel has low skin-irritation potential. AB - BACKGROUND: Adapalene is a new naphthoic acid derivative with potent retinoid and antiinflammatory properties, developed for the topical treatment of acne vulgaris. OBJECTIVE: We compare the cutaneous safety of adapalene in different gel vehicles with tretinoin 0.025% gel. METHODS: A total of 42 healthy human subjects were enrolled in two randomized, double-blind, controlled, intraindividual studies. In the first study (study A), adapalene aqueous 0.03% and 0.1% gels were evaluated for their 21-day cumulative irritation potential compared with vehicle alone, patch alone, and tretinoin 0.025% gel under occlusion. In the second study (study B), adapalene aqueous (0.03% and 0.1%) gels and adapalene alcoholic (0.03% and 0.1%) gels were evaluated for their 5-day cumulative irritation potential compared with their respective vehicles and tretinoin 0.025% gel. Transepidermal water loss (TEWL) was measured daily at each visit. RESULTS: In study A, adapalene had a slight irritation potential that was in the same range as the gel vehicle and the patch alone, whereas tretinoin 0.025% gel was a severe irritant. In study B, no irritation was seen with either adapalene aqueous gels or adapalene gel vehicles or patch alone. The adapalene alcoholic gels were slightly irritating, and tretinoin gel produced intense irritation reactions in the majority of subjects. TEWL increased fourfold at the tretinoin site but remained unchanged at all adapalene sites. CONCLUSION: Adapalene 0.1% gel was significantly less irritating than tretinoin 0.025% gel. PMID- 9204087 TI - Split-face comparison of adapalene 0.1% gel and tretinoin 0.025% gel in acne patients. AB - BACKGROUND: Adapalene is a new naphthoic acid derivative developed for the topical treatment of acne vulgaris. OBJECTIVE: We compared the skin tolerance of adapalene 0.1% gel with tretinoin 0.025% gel in subjects with acne. METHODS: Fifteen acne patient volunteers were enrolled in this investigator-masked, left right comparison, randomized, controlled, intraindividual study. Adapalene 0.1% gel and tretinoin 0.025% gel were applied once a day to one half-face by the volunteers for 14 consecutive days. Clinical signs (erythema, desquamation, papules, vesicles, edema) and subjective symptoms (tightness, pruritus, burning) were evaluated and scored daily except on weekends. RESULTS: Adapalene 0.1% gel was better tolerated than tretinoin 0.025% gel. The overall mean score calculated from all features combined was significantly higher with tretinoin gel than with adapalene gel (p = 0.002). CONCLUSION: Adapalene 0.1% gel was significantly less irritating than tretinoin 0.025% gel when tested in acne patients. PMID- 9204088 TI - Skin tolerance of adapalene 0.1% gel in combination with other topical antiacne treatments. AB - BACKGROUND: Adapalene (Differin gel) is a new naphthoic acid derivative developed for the topical treatment of acne vulgaris. OBJECTIVE: We assessed, in healthy volunteers, the skin irritancy potential of three combinations, each including adapalene 0.1% gel and one topical marketed antiacne product. METHODS: Twenty five healthy volunteers were enrolled in a 21-day cumulative irritancy study performed in a double-blind, randomized, controlled, intraindividual design. Five days a week, the three materials (benzoyl peroxide, clindamycin phosphate, and erythromycin) were applied in a nonocclusive manner either alone or in combination with adapalene gel on seven cutaneous sites on the upper back. Adapalene was applied in the evening whereas the three other materials were applied in the morning. Irritation was evaluated and scored daily except on weekends. RESULTS: All materials were well-tolerated when tested alone. The combinations of adapalene 0.1% gel and either benzoyl peroxide, clindamycin phosphate, or erythromycin were also well-tolerated. The mean cumulative irritancy indices indicated that all three combinations were nonirritating. CONCLUSION: Under the conditions of the study, all tested treatments alone or in combination appeared nonirritating. PMID- 9204089 TI - Adapalene 0.1% gel is better tolerated than tretinoin 0.025% gel in acne patients. AB - BACKGROUND: Adapalene is a new naphthoic acid derivative developed for the topical treatment of acne vulgaris. OBJECTIVE: We describe the results of a combined safety analysis of two multicenter trials conducted in the U.S. and Europe in which adapalene 0.1% gel was compared with tretinoin 0.025% gel in the treatment of mild to moderate acne vulgaris. METHODS: A total of 591 acne patients were enrolled in these investigator-masked, randomized, controlled, parallel group studies. In the two studies, each patient was randomly assigned to receive topical adapalene 0.1% gel or tretinoin 0.025% gel once daily at bedtime, for 12 weeks. In addition to assessments of efficacy and facial skin tolerance, data on adverse events were recorded at each visit or at any other time the patient reported problems. We extracted data concerning adverse reactions (i.e., adverse events judged to be related to the study treatment) from both studies and combined the results to obtain a global comparison of safety of the two products. RESULTS: A total of 15 of 296 patients (5.1%) reported 19 adverse reactions in the adapalene-treated groups, compared with 27 of 295 patients (9.1%) reporting 39 adverse reactions in the tretinoin-treated groups (p < 0.05). The number of patients discontinuing the study because of adverse events was approximately twice as low with adapalene (1.3% compared with 2.4%). Most adverse reactions for both products were related to skin irritation. No systemic adverse reactions were reported. CONCLUSION: The results of these two multicenter clinical studies indicate that adapalene gel is better tolerated than tretinoin gel. PMID- 9204090 TI - Skin distribution and pharmaceutical aspects of adapalene gel. AB - BACKGROUND: The formulation of topical dermatologic products must take into account efficacy, safety, and patient compliance. OBJECTIVE: We describe how an aqueous gel containing adapalene, a novel naphthoic acid derivative was selected for the topical treatment of acne. METHODS AND RESULTS: Dose selection was made using established in vivo models. In the Rhino mouse the comedolytic effect of adapalene 0.1% gel was similar or superior to that of a reference formulation containing 0.025% tretinoin. Qualitative distribution of adapalene after topical application was assessed on cryosections of human skin. They showed rapid penetration of adapalene into the pilosebaceous unit. A liberation/penetration study demonstrated that significant quantities of adapalene were present in epidermis and dermis, but only 0.01% of the applied dose penetrated through the skin. CONCLUSION: This formulation should result in effective and well-tolerated treatment of acne vulgaris with good patient compliance. PMID- 9204092 TI - Evaluation of antidotes: activities of the International Programme on Chemical Safety. AB - Important developments concerning the role of antidotes in managing poisoning cases have taken place in recent decades due to new toxicodynamic and toxicokinetic studies and to growing international concern regarding the effectiveness of antidotes. A number of activities are carried out by the International Programme on Chemical Safety which aim to: (1) evaluate their effectiveness in clinical practice, (2) disseminate evaluated information, and (3) promote the availability of useful antidotes. The International Programme on Chemical Safety has undertaken the preparation of Antidote Monographs that summarize and assess the clinical use, mode of action, effectiveness, and other evaluated information, and a consolidated International Programme on Chemical Safety List of Antidotes that classifies antidotes and related drugs by their clinical effectiveness and urgency of need. A chart of Antidote Dosages, with information concerning the recommended antidotes and their indications, is being prepared, and the Availability of Antidotes in different countries is being surveyed. Further International Programme on Chemical Safety initiatives are also being undertaken in the area of antidotes and clinical toxicology in order to examine particular issues. The International Programme on Chemical Safety INTOX Project and related activities provide powerful tools for multicenter studies, but such research faces continuing financial and regulatory difficulties. Twinning arrangements between scientists from different parts of the world are being promoted to enhance the capabilities of evaluating treatment procedures and to compare clinical data. International organizations have important aims: to promote adequate and appropriate regulations and increase antidote availability, to establish international consensus and to increase interest in co-operative research. Cooperation with scientific bodies is essential in supporting these aims. PMID- 9204091 TI - Clinical efficacy and safety comparison of adapalene gel and tretinoin gel in the treatment of acne vulgaris: Europe and U.S. multicenter trials. AB - BACKGROUND: Adapalene is a new chemical entity that exhibits tretinoin-like activities in the terminal differentiation process. OBJECTIVE: We evaluated a dose range effect of two concentrations of adapalene gel as acne treatment and compared adapalene 0.1% gel with tretinoin 0.025% gel in the treatment of acne patients in two large multicenter studies. METHODS: Multicenter, investigator masked, parallel group studies including 89 acne patients in the dose range study and 591 patients in the concurrent controlled studies were conducted. RESULTS: Adapalene gel 0.1% was significantly more effective in treating acne lesions than 0.03% adapalene gel. Adapalene gel 0.1% was significantly more effective than 0.025% or tretinoin gel in one study and of the same effectiveness in the other study. Adapalene gel was always better tolerated than tretinoin gel. CONCLUSION: Adapalene 0.1% gel is a safe and effective treatment of acne vulgaris. PMID- 9204093 TI - Therapeutic, toxic, and lethal concentrations in human fluids of 90 drugs affecting the cardiovascular and hematopoietic systems. AB - BACKGROUND: Drugs affecting the cardiovascular and hematopoietic systems are frequently involved in poisoning. As a continuation of our previously published study about the concentrations of drugs of abuse, we have compiled published data about these drugs and subjected them to selection and unification on the basis of conservative criteria and our own experience. RESULTS: A compilation of the concentrations of 90 drugs affecting heart, circulation, blood or hematopoietic organs, in whole blood, serum/plasma, and urine, corresponding to therapeutic, toxic or lethal concentrations is given. Although the interpretation of the concentrations is a complex and difficult problem, the presented table can be helpful in interpretation from the actual concentrations of this group of drugs encountered in clinical, toxicological and forensic cases. PMID- 9204094 TI - Characterization of fatal beta blocker ingestion: a review of the American Association of Poison Control Centers data from 1985 to 1995. AB - OBJECTIVE: To characterize beta blocker-related deaths. METHODS: This is a retrospective review of beta blocker-related exposure data and fatality case abstracts reported to the American Association of Poison Control Centers Toxic Exposure Surveillance System during the 11 year period, 1985 to 1995. Historical and laboratory data were used to determine those fatalities which resulted primarily from beta blocker intoxication. RESULTS: Of 52,156 reported beta blocker exposures, 164 were fatal. In 38 cases, beta blockers were implicated as the primary cause of death. Propranolol was responsible for the greatest number of exposures (44%) and implicated as the cause of death in a disproportionately high percentage of fatalities (71%). Patients were generally young women; 63% were female and 92% were less than 50 years old. The dysrhythmias most often noted in fatal cases were bradycardia and asystole. Cardiopulmonary arrest did not develop until patients were in the care of health care personnel in 59% of cases. Though glucagon was initiated more often than any other intervention in fatal intoxications (83%), optimal dosing and maintenance infusions appear to have been underutilized. CONCLUSIONS: The predominance of fatalities associated with propranolol compared to other beta blockers reflects both its greater frequency of use over the time period studied and its greater toxicity. Since 59% developed. cardiac arrest after reaching health care personnel, further study should focus on identifying medical intervention that can reduce mortality in this group. PMID- 9204095 TI - Prospective multicenter evaluation of tramadol exposure. AB - BACKGROUND: Tramadol is a novel analgesic possessing both opiate and noradrenergic effects. Its low potential for abuse suggests increasing use, but there are limited data on the toxicity in overdose. METHODS: Multicenter prospective case series. All exposures from October 1995 through August 1996 reported to seven Poison Centers were evaluated. RESULTS: There were 126 cases of which 87 were tramadol alone. Of the tramadol alone cases, 51 were female (59%). Age ranged from 1 to 86 y with a mean and median of 26.8 y (SD 17.2) and 25 y, respectively. There were 15 cases of children less than 6 years old. Symptoms reported with overdose were: lethargy 26 (30%), nausea 12 (14%), tachycardia 11 (13%), agitation 9 (10%), seizures 7 (8%), 4 each (5%) of coma and hypertension, and respiratory depression 2 (2%). All seizures were brief. Naloxone reversed sedation and apnea in 4 of 8 patients. One patient experienced a seizure immediately after administration of naloxone. Other treatments were: diazepam (3 patients), and phenytoin, lorazepam and nifedipine (1 patient each). Tramadol 500 mg was the lowest dose associated with seizure, tachycardia, hypertension or agitation while 800 mg was the lowest dose associated with coma and respiratory depression. Urine drug screens performed on 19 patients were negative for opiates. All symptomatic cases exhibited effects within 4 h of ingestion. Mean hospital stay was 15.2 h (range 2-96 h, SD 15.8). Nineteen patients were admitted to an intensive care unit with a mean stay of 25 h (SD 20). DISCUSSION: Much of the toxicity in tramadol overdose appears to be attributable to the monoamine uptake inhibition rather than its opioid effects. Agitation, tachycardia, confusion and hypertension suggest a possible mild serotonin syndrome. No arrhythmias beyond tachycardia were seen. CONCLUSION: This study suggests significant neurologic toxicity from tramadol overdose. Serious cardiovascular toxicity was not seen. PMID- 9204096 TI - Ethanol potentiates the depressant effects of cocaine in human fetal myocardium in vitro. AB - BACKGROUND: Cocaine and ethanol use is widespread in our society and is not uncommon in pregnant women. Previous work has demonstrated that acute exposure to cocaine produced significant effects on the configuration of human fetal myocardial action potentials and contractility in vitro. It has been hypothesized that these target-specific effects of cocaine may provide a plausible mechanism to account for fetal arrhythmia or sudden fetal death in utero. OBJECTIVE: This study was conducted to determine if treatment with a low concentration of ethanol (200 mg/L) would predispose human fetal myocardium to cocaine-induced toxicity in vitro. METHODS: Fetal hearts (12-14 weeks) were obtained at the time of elective abortion and transported to the laboratory in cold physiological salt solution. The force of muscle contractions and transmembrane potentials of ventricular walls were studied during external electrical stimulation in a specially constructed superfusion chamber. RESULTS: When a concentration of cocaine (200 micrograms/L physiological salt solution) that singly produced only modest myocardial depression was used in combination with a concentration of ethanol which alone produced nonsignificant changes, marked depression and block of action potentials and contractility resulted. CONCLUSION: The combined use of ethanol and cocaine produces fetal myocardial depression which is greater than that predicted from the effects of these chemicals individually and may have significant in utero implications. PMID- 9204097 TI - The lead concentration of reconstituted infant formula. AB - OBJECTIVE: To characterize practices of infant formula reconstitution and to measure the lead concentration of the home-prepared reconstituted infant formula. METHODS: Convenience sample of metropolitan Boston infants less than 9 months of age who were being evaluated in an urban pediatric emergency department and had home-prepared reconstituted infant formula available. A questionnaire was administered to gather demographic information and details of formula preparation. A 30-90 mL sample of home-prepared reconstituted infant formula was collected in an acid-washed, distilled water-rinsed glass tube, then analyzed for lead by atomic absorption spectrophotometry (precision +/- 2 micrograms/L or 2 ppb). RESULTS: Forty infants were evaluated. Mean (SD) chronological age was 112 +/- 78 d. Reported daily reconstituted infant formula daily volume was 870 +/- 300 mL. Two of the 40 samples (5% [95% CI: 2.0, 18.2%]) had lead concentrations above 15 micrograms/L, the current action level for safe water according to the Environmental Protection Agency. The two samples with lead concentrations of 17 and 70 micrograms/L were prepared using cold tap water (water run for 5 and 30 sec, respectively) drawn from the plumbing of houses greater than 20 years old. CONCLUSION: These data suggest that the use of infant formulas which require reconstitution may present inadvertent lead hazards to young infants. Pediatricians should provide education about these potentially hazardous practices to parents who use these formulas. PMID- 9204098 TI - Concentrations of blood and hair mercury and serum PCBs in an Ojibwa population that consumes Great Lakes region fish. AB - OBJECTIVE: This paper describes an exposure assessment of an American Indian population using blood and hair samples as indicators of mercury and polychlorinated biphenyl exposure from the consumption of fish taken from the Great Lakes region. METHODS: Questionnaires regarding fish consumption were completed by 89 Ojibwa tribal members. Mercury concentrations were determined in human hair and blood samples, and polychlorinated biphenyl concentrations were determined in serum. RESULTS: Fish were consumed at the highest rates in April, May, June, and July. Lake trout, whitefish, and walleye were the preferred fish consumed by 91.4% of the respondents. Concentrations of blood mercury were all below 55 micrograms/L (ppb), while concentrations of mercury in hair were all less than 3 mg/L (ppm). Hair mercury concentrations were correlated with the previous year's fish consumption (p = .05). Dental amalgams and blood mercury concentrations were also significantly correlated (p < .002). Serum polychlorinated biphenyl concentrations, determined as the sum of 89 congeners, were all below 9.6 ppb total polychlorinated biphenyls. Subject age and total serum polychlorinated biphenyls were correlated (p < .001). CONCLUSIONS: The concentrations of mercury and polychlorinated biphenyls in this Ojibwa population were relatively low, but several individuals were identified as having elevated concentrations and additional testing may be warranted. Since the accumulation of contaminants was related to fish consumption and age, a long-term monitoring program that assesses chronic exposure to fish diets would be beneficial. PMID- 9204099 TI - A retrospective study of poisoning in Tehran. AB - OBJECTIVE: To examine the causes and mortality of poisoning in Tehran. METHODS: The 7000 poisoning cases referred to Loghman-Hakim Hospital in Tehran over six months in 1994 were evaluated retrospectively. RESULTS: The overall female to male ratio was 1.8:1. Most poisonings occurred in the age range 2-6 y for children and 21-40 y for adults. Oral ingestion was the most common route of intoxication. In children, boys had a higher frequency of poisonings than girls. Most cases of children were referred to the hospital between 8 am and 8 pm. In adults referred to the hospital, there was little diurnal variation in poisoning presentations. In adults, drugs were the most common cause of intoxication (60.2%). Of these, benzodiazepines (24.5%) were the most frequent, followed by antidepressants (20.5%) and analgesics (18%). Pesticide and opiate intoxications were also commonly observed. In children, after drugs (32.1%), hydrocarbons were the most frequent cause of poisoning (19.2%). Pesticide poisonings were most often fatal (19.2%), followed by barbiturates (18.6%) and opiates (16.2%). Organophosphate insecticides were responsible for 57% of total pesticide poisoning cases. Of the deaths, 87.5% were attributed to suicide. CONCLUSION: The majority of poisoning cases in adults occur intentionally and in children accidentally. PMID- 9204100 TI - Tacrolimus (FK 506) overdose: a report of five cases. AB - INTRODUCTION: Tacrolimus (FK 506), a potent anti-T cell agent, has been shown to be effective in preventing the rejection of transplanted organs. Published research on tacrolimus has focused on effects associated with therapeutic use. Virtually no literature addresses the acute toxicity or the management of tacrolimus overdose. We report five cases of acute overdose with tacrolimus. CASE REPORTS: A 2-year-old female with no prior medical history ingested 10 mg of tacrolimus. She remained asymptomatic. A 2-year-old female with a history of multiple visceral organ transplants ingested 11 mg of her tacrolimus. She was admitted to the hospital and activated charcoal was administered. Her renal function was monitored and no changes were noted in a 24 h period. She was discharged. A 29-year-old male renal transplant patient took 150 mg of tacrolimus. He recovered with only a minimal creatinine elevation. A 23-year-old heart and lung transplant patient ingested 375 mg of tacrolimus. She had no effects from the overdose. A 34-year-old female experienced an acute/chronic overdose of 7-9 mg and remained asymptomatic. DISCUSSION: Tacrolimus is a neutral macrolide antibiotic that is extracted from the fermentation broth of the soil fungus Streptomyces tsukubaensis. Chronic oral dosing has been associated with numerous side effects. Although these patients ingested significant doses of tacrolimus, they suffered few toxic manifestations associated with tacrolimus. CONCLUSION: Little information is available regarding acute tacrolimus overdosage. In this small series of patients, tacrolimus did not produce acute physiologic incapacitation. PMID- 9204101 TI - Isofenphos poisoning: prolonged intoxication after intramuscular injection. AB - We report a unique case of attempted suicide by intramuscular injection of the organophosphate isofenphos which resulted in a muscarinic and nicotinic syndrome lasting 15 days and requiring prolonged mechanical ventilation and hospitalization. The patient, who demonstrated no signs of delayed polyneuropathy on hospital day 25, subsequently died of pneumonia. Toxicological investigations showed isofenphos plasma decay and confirmed the intramuscular route of poisoning. We believe continuous isofenphos absorption resulted in the prolonged intoxication observed in this patient. PMID- 9204103 TI - Hydroxocobalamin analysis and pharmacokinetics. PMID- 9204102 TI - Massive ingestion of the herbicide 2-methyl-4-chlorophenoxyacetic acid (MCPA). AB - OBJECTIVE: To report the course of a massive ingestion of the herbicide 2-methyl 4-chlorophenoxyacetic acid or MCPA (4-chloro-2-methyl phenoxy acetic acid) and to correlate plasma 2-methyl-4-chlorophenoxyacetic acid levels with symptoms of intoxication and treatment. CASE REPORT: After intentional ingestion of the herbicide, 2-methyl-4-chlorophenoxyacetic acid, a young man suffered burning in his mouth, spasmodic pain in the extremities and a severe hypotensive crisis. Plasma 2-methyl-4-chlorophenoxyacetic acid concentration was 546 mg/L two hours after ingestion. Therapy by forced diuresis was ineffective until the urine was alkalinized (Day 4). This resulted in a rapid decline of the plasma 2-methyl-4 chlorophenoxyacetic acid level to 6 mg/L and recovery of the patient. PMID- 9204104 TI - Driving under the influence of acetone. PMID- 9204105 TI - Sequential computerized tomography changes and related final outcome in severe head injury patients. AB - The authors analysed the serial computerized tomography (CT) findings in a large series of severely head injured patients in order to assess the variability in gross intracranial pathology through the acute posttraumatic period and determine the most common patterns of CT change. A second aim was to compare the prognostic significance of the different CT diagnostic categories used in the study (Traumatic Coma Data Bank CT pathological classification) when gleaned either from the initial (postadmission) or the control CT scans, and determine the extent to which having a second CT scan provides more prognostic information than only one scan. 92 patients (13.3% of the total population) died soon after injury. Of the 587 who survived long enough to have at least one control CT scan 23.6% developed new diffuse brain swelling, and 20.9% new focal mass lesions most of which had to be evacuated. The relative risk for requiring a delayed operation as related to the diagnostic category established by using the initial CT scans was by decreasing order: diffuse injury IV (30.7%), diffuse injury III (30.5%), non evacuated mass (20%), evacuated mass (20.2%), diffuse injury II (12.1%), and diffuse injury I (8.6%). Overall, 51.2% of the patients developed significant CT changes (for worse or better) occurring either spontaneously or following surgery, and their final outcomes were more closely related to the control than to the initial CT diagnoses. In fact, the final outcome was more accurately predicted by using the control CT scans (81.2% of the cases) than by using the initial CT scans (71.5% of the cases only). Since the majority of relevant CT changes developed within 48 hours after injury a pathological categorization made by using an early control CT scan seems to be most useful for prognostic purposes. Prognosis associated with the CT pathological categories used in the study was similar independently of the moment of the acute posttraumatic period at which diagnoses were made. PMID- 9204106 TI - Unilateral laminotomy for bilateral decompression of lumbar spinal stenosis. Part I: Anatomical and surgical considerations. AB - A unilateral laminotomy for bilateral access to the lumbar spinal canal was investigated in human cadaver spine specimens to test its practicability in the treatment of spinal stenosis. Micro-surgical decompression was performed by partial resection of the ipsilateral facet, the medial portion of the laminar arch, the contralateral facet and by complete removal of the ligamentum flavum. Anatomical, radiological and morphometrical studies on 4 adult cadaver spine specimens have proved the feasibility of this unilateral approach. Complete bilateral flavectomy and partial bilateral facetectomy were the essential surgical steps for an adequate operative decompression. PMID- 9204107 TI - Unilateral laminotomy for bilateral decompression of lumbar spinal stenosis. Part II: Clinical experiences. AB - The surgical aim in the treatment of symptomatic lumbar spinal stenosis is the relief of the patient's complaints by an adequate neural decompression. Unilateral laminotomy and bilateral spinal canal decompression represents such a safe, effective and minimally invasive surgical method. This technique has been successfully used in the operative treatment of 29 patients with symptomatic mono or multisegmental lumbar stenosis. There was no surgically induced neurological deterioration. In one patient, an inadvertent dural tear occurred, and due to unchanged symptoms another patient with a multisegmental stenosis had to be re operated on at an additional level. Postoperatively, 25 of the 27 patients with neurogenic claudication (93%) demonstrated a marked improvement of the walking distance. The follow-up of 25 patients (mean follow-up time was 18 months) demonstrated an excellent result without pain in 7 patients (28%); a good outcome with mild residual pain, but a normal working capacity in 15 patients (60%); and a fair outcome with unchanged postoperative low-back pain but markedly improved working capacity and walking distance in 3 patients (12%). Postoperative morphometric evaluation as well as the clinical improvement of the patient's symptoms clearly demonstrated that bilateral ligamentectomy and recess decompression were adequately and successfully achieved via unilateral approach. PMID- 9204108 TI - Extensive damage to the end-plates as a complication of laser discectomy. An experimental study using an animal model. AB - To date, there have been no reports of experiments demonstrating the effects of neodymium:YAG laser (Nd:YAG laser) on the vertebral end-plates. In this study the effect of Nd:YAG laser on end-plates was examined in 32 guinea pigs which were randomly divided into two groups. The first group was the control group, the second one the Nd:YAG laser group. All animals had experimental disc degeneration at three levels. Re-exploration was performed two months after the surgical ventral disc herniation. In the second group the same procedure was performed but at the end of the re-exploration, Nd:YAG laser irradiation of the degenerated discs was done. The wounds in both groups were closed again. Two months later all animals were sacrificed for histological and biochemical analysis. The cervical spine was excised en bloc and the overlying muscles were removed. Determination of hydroxyproline was done colorimetrically in the specimens harvested from each of these groups. Light microscopy was undertaken to evaluate the extent of morphological changes. The differences observed between the two groups were statistically significant (p < 0.05). From the results of this study, there is the question whether the Nd:YAG laser is a useful instrument in neurosurgery. Therefore, it remains to be proven whether or not this is of real benefit in the treatment of patients with degenerated disc disease. PMID- 9204109 TI - Muslin induced granuloma following wrapping of intracranial aneurysms: the role of infection as an additional precipitating factor. Report of two cases and review of the literature. AB - Two patients who developed what appeared to be a granulomatous reaction following muslin wrapping of unclipped aneurysms are reported. They presented with cranial nerve palsies and at operation were found to have an abscess around the wrapped aneurysms. In one of these two patients Staphylococcus epidemidis was isolated from the pus. This, together with further evidence from reported cases in the literature, would suggest that infection may play an additional role at least in some cases in the onset of a foreign-body granulomatous reaction seen following wrapping of aneurysms. PMID- 9204110 TI - Angiographically unrecognized microaneurysms: intraoperative observation and operative technique. AB - Twenty operated cases of angiographically unrecognized microaneurysm (AUM) have been analysed with special reference to intra-operative observations and clipping technique. Among the patients with intracranial aneurysms that the authors' facility has operated upon, the incidence of asymptomatic incidental AUM that was 2 mm or smaller amounted to 3.7%. Thirteen cases of AUM were found on the middle cerebral artery; four AUMs arose from the M1 portion, four from the bifurcation, and five from the second bifurcation. Sixty percent of AUMs were recognized on the parent arteries of ruptured aneurysms. In 90% of cases the AUMs were broad based in shape and in 70% of cases exhibited a thin-walled neck and a thin-walled fundus. Intra-operative findings revealed four reasons why AUMs were not visible in the pre-operative angiograms: (1) the AUM was sandwiched between two arteries; (2) the AUM was completely hidden by a contiguous large or giant aneurysm; (3) the AUM was diagnosed by pre-operative angiogram as a bleb of the contiguous aneurysm; (4) the AUM was not visible on angiograms because the height of the AUM was extremely low. Twelve cases of AUM were successfully clipped using four different clipping techniques; (1) clipping parallel to the bifurcation in four, (2) clipping parallel to the parent artery in four, (3) pinch-clipping in two of the cases, and (4) cross clipping in two of the cases. The other eight cases were wrapped and coated. AUMs may be present during the direct operation of intracranial aneurysms and in intravascular surgery. Neurosurgeons and neuroradiologists need to explain the possible existence of AUMs to patients and their families. PMID- 9204111 TI - A prospective study of microvascular decompression for trigeminal neuralgia. AB - In a prospective study of 25 patients with trigeminal neuralgia (TN), we examined the results of microvascular decompression (MVD). Initial pain relief was complete in 22 patients and partial in one. There were two primary failures. After a median observation time of 38 months, 20 of the 22 patients still were completely free of pain, and one patient reported then 50% pain relief. A vascular compression of the trigeminal root was found intra-operatively in 23 patients. No serious complications occurred. Minor but bother-some dyaesthesias were reported by two patients (8%). The results were satisfactory when compared to other MVD studies. PMID- 9204112 TI - MIB1 immunopositivity is associated with rapid regrowth of pituitary adenomas. AB - Pituitary adenomas are generally regarded as benign tumours, but they may recur. We identified eight patients with pituitary adenomas that showed rapid regrowth within 2 years of initial surgery. We estimated the percentage of cells in each specimen that showed positive immunostaining for MIB1 (a novel anti-Ki-67) and compared the values to those of 40 adenomas that showed no regrowth. The mean MIB1 index for 40 adenomas that showed no evidence of regrowth was 0.19 +/- 0.06%. This was significantly (p < 0.0001) lower than that for adenomas that showed rapid regrowth (1.27 +/- 0.31%), based on the initial resected specimens. Immediately after detection of rapid regrowth and in adenomas that were resistant to bromocriptine or irradiation, the MIB1 index was always greater than 1.0%. Most patients with rapidly regrowing adenomas were well controlled by radiation therapy. Our results suggest that a MIB1 index greater than 1.0% may be a useful predictor of rapid regrowth of pituitary adenomas and may be useful for planning of therapy. PMID- 9204113 TI - Electrophysiologic target localization in posteroventral pallidotomy. AB - The current interest in stereotactic posteroventral pallidotomy (PVP) for treating Parkinson's disease and the variability of published results have raised questions regarding techniques for target localization. In our technique the probe is guided to the optimum target at the most ventral pallidum and ansa lenticularis by macroelectrode stimulation of the internal capsule and optic tract from within the globus pallidus, with the thresholds providing a relative measure of the electrode proximity to these structures. We have characterized these localizing macroelectrode stimulation parameters in 57 posteroventral pallidotomies with consistent anatomic lesion placement, excellent outcome, and no complications. Using a 1.8 x 2.0 mm radiofrequency electrode for macroelectrode stimulation (RFG-3C, Radionics Inc.), minimum voltages (thresholds) to activate motor (at a frequency of 2 Hz) or visual (at a frequency of 100 Hz) responses as well as impedance measurements were obtained at the final target (Tf) and at distances proximal to Tf along the electrode trajectory. The visual and motor threshold voltages at Tf via our standard approach angles (50 degrees above base plane, 20 degrees from the sagittal plane), had a range of 1.0 to 1.5 V, and 2.0 to 3.5 V respectively. We also found that as the probe approaches Tf there is a significant decrease in voltage thresholds for motor (P < .0001) and visual (P < .0001) responses in an individual patient indicating that the probe is converging on these structures. Increases in impedance between Tf, 2-3 mm, and 4-5 mm proximal to Tf were also statistically significant (P < .0001). Microelectrode recording to electrophysiological neuronal activity at various points along the trajectory towards the target showed distinct firing patterns providing identification of the globus pallidus externus and internus, ansa lenticularis, and optic tract. Macroelectrode electrophysiological stimulation within the target volume, inducing threshold responses in the internal capsule and optic tract, provides for accurate localization of the most effective PVP target in the ansa lenticularis. In unresponsive patients, the utilization of microelectrode recording for the identification of the pallidal borders and the optic tract improves safety. PMID- 9204114 TI - Gamma knife pallidotomy: case report. AB - We report a case of gamma knife pallidotomy resulting in a permanent contralateral homonymous hemianopsia and transient contralateral hemiparesis with some improvement in contralateral parkinsonian symptoms. This case illustrates the risk of gamma knife pallidotomy which precludes physiologic target localization and can subject structures surrounding the target to a significant radiosurgical dose. Until noninvasive physiologic target localization is available gamma knife pallidotomy and thalamotomy should be limited to patients with an unacceptably high risk for stereotactic percutaneous thermocoagulation. PMID- 9204115 TI - Osteoregenerative lateral suboccipital craniectomy using fibrin glue. AB - This report describes a simple technique of cranioplasty for suboccipital craniectomy and the results of a clinical study to assess the effects of fibrin glue on regeneration of the skull. Cranioplasty using a mixture of autologous bone chips and human allogenic fibrin glue was performed in 31 patients who received lateral suboccipital craniectomy. Long-term observations with three dimensional CT showed satisfactory reconstruction of the mastoid-occipital bone plate in 25 patients (81%); among them, a nearly complete reconstruction of the occipital bone (plate) was found in 14 patients. Regeneration of the bone began 6 months after surgery on the inner surface, adjacent to the dura mater. In conclusion, the present technique provides a new simple method to restore an autologous bone plate in a cranial defect made by piecemeal craniectomy. PMID- 9204116 TI - Efflux of gamma-aminobutyric acid caused by changes in ion concentrations and cell swelling simulating the effect of cerebral ischaemia. AB - The relationships among ischaemic GABA efflux from brain tissue and extracellular and intracellular concentrations of sodium, chloride and potassium ions were investigated by means of 1) transverse hippocampal slices from rat and 2) functional expression of a high affinity GABA transporter in Xenopus oocytes. Brain slices were incubated for 20 min in medium where extracellular sodium and chloride were substituted with impermeant ions. Isethionate (Iseth) substitution for chloride generated a 7-fold increase in GABA efflux. Choline (Chol) but not N methyl-D-glucamine (NMDG) substitution for sodium likewise increased GABA efflux. Reducing the osmolarity of the medium by decreasing both sodium and chloride concentrations (Hyp) increased GABA efflux 3-fold. This release was blocked by mannitol (Man). Blocking sodium channels with 1 microM of tetrodotoxin (TTX) also increased the release 3-fold. Energy deprivation (ED) increased the GABA release 50-fold. ED/Iseth left the release unchanged, ED/Chol increased the GABA efflux by 23%, whereas ED/NMDG reduced the release by 41%. Adding mannitol did not block the ED-evoked release, whereas TTX reduced it by 52%. Release of preloaded [3H] GABA from oocytes expressing the GAT-1 GABA transporter was then examined. Depolarisation by current injection or 100 mM extracellular K+ did not increase GABA release. Sodium chloride injection, however, caused membrane depolarisation and a 100-fold increased GABA efflux from the oocytes. This release was blocked when the osmolarity was increased extracellularly by adding mannitol. These results show that 1) TTX releases GABA from brain tissue but blocks release during ED, 2) the high affinity GABA carrier must be altered in order to reverse, 3) ischaemic GABA release is sodium independent, and is modulated by large cations, 4) mannitol blocks the reversal of high affinity carriers in oocytes, but the release from brain slices during ED is unaffected. Taken together, the results suggest that ischaemic release of GABA from brain tissue does not occur by means of reversed high affinity carriers alone, but rather that it is controlled by more complex mechanisms. PMID- 9204117 TI - The time course and regional variations of lipid peroxidation after diffuse brain injury in rats. AB - Free radicals are generated after head injury. These radicals rapidly react with polyunsaturated fatty acids in the cell membrane and cause membrane destruction. This process is called lipid peroxidation. Malondialdehyde (MDA) is one of the end products of lipid peroxidation, and it is a frequently used indicator of lipid peroxidation in biological tissues. Using a diffuse head injury animal model, we studied the time course of lipid peroxidation in different regions of injured rat brains. In the present study, the MDA levels were 36.7%, 41.8%, and 35.1% greater than sham at one hour after injury at the frontal, parietal, and brain stem, respectively (p < 0.0001). The MDA levels in these regions continued to increase and peaked a 4 hours after the injury. The levels slowly decreased, and by 24 hours, they were still significantly higher than the sham control's. The elevation of MDA levels was less in the striatum and the temporal regions at one hour. They were 16.9% and 13.3%, respectively (p < 0.002). The MDA levels in these two regions continued to increase even after 4 hours of injury, but the degree of elevation never exceeded 35%. The results demonstrate that there is an immediate, posttraumatic burst of MDA production, suggesting the formation of free radicals after diffuse head injury. Even though all the regions sampled show the same effect, certain regions are less affected by this diffuse head injury animal model. PMID- 9204118 TI - Loss and apoptosis of smooth muscle cells in intracranial aneurysms. Studies with in situ DNA end labeling and antibody against single-stranded DNA. AB - Pathological specimens were collected from 14 unruptured and 13 ruptured aneurysms at the time of clipping and studied in order to assess the underlying mechanism of rupture by investigating degeneration of the aneurysmal wall and possible involvement of apoptosis. Immunohistochemistry with anti-actin antibody showed few smooth muscle cells in the ruptured aneurysms and replacement of the muscularis layer by a fibro-hyalin tissue. However, at least one layer of smooth muscle cells was clearly observed in the unruptured aneurysms. Thus, smooth muscle cells in the wall of the ruptured aneurysms were much more degenerated than those in the wall of unruptured aneurysms. In addition, unruptured aneurysms with an angiographically smooth wall showed well-layered positive staining for anti-smooth muscle actin antibody while those with irregular shapes rarely reacted. We found, for the first time, evidence of DNA fragmentation in the aneurysmal wall. Apoptotic bodies were detected by means of a terminal transferase (TdT)-mediated dUTP biotin nick end labelling technique (TUNEL) and an anti-single-stranded DNA antibody in 54% (7/13) of the ruptured aneurysms. In contrast, apoptotic bodies were found in only 7% (1/14) of the unruptured cases. These results suggest that apoptotic cell death might be involved in the rupture of aneurysms. PMID- 9204119 TI - Primary embryonal cell carcinoma of cerebellopontine angle. PMID- 9204120 TI - MR imaging of multiple cerebral infarctions in angiotropic lymphoma (neoplastic angio-endotheliosis). PMID- 9204121 TI - Angiographically occult anomalous ophthalmic artery arising from the anterior cerebral artery. PMID- 9204122 TI - Permanent postoperative anosmia: a hitherto undescribed complication following surgery of the posterior cranial fossa in the sitting position. AB - Although the sitting position offers advantages for posterior fossa surgery, it is accompanied by complications such as air embolism and pneumatocephalus. Subdural and epidural haematomas are less common postoperative complications of posterior fossa surgery. To the best of our knowledge, however, anosmia is not a known sequela of surgery in the sitting position. It has been described following aneurysm surgery in the rostral part of the circle of Willis and is, of course, well known in traumatic brain injury. PMID- 9204123 TI - Protein arrays: concepts and subjects. AB - To adapt proteins, the materials in life, for use as materials in science and technology, we focused not only on the biological aspects (functional aspects) but also on the material aspects as matter (structural and physical aspects). Engineering with protein arrays will develop under such consideration and advance toward stable devices made of protein molecules. The protein arrays with 2D crystalline order provide a primary model of macroscopic protein-based devices. The combination of protein engineering, the leading edge of life science, and array engineering, the leading edge of materials science, will provide clues to the controlled integration of protein molecules to a form of functional supramolecules on proper surfaces. PMID- 9204124 TI - Lateral forces acting between particles in liquid films or lipid membranes. AB - To investigate the mechanism of formation of 2D arrays of protein macromolecules in liquid films we carried out model experiments with micron-sized latex particles. The direct observations revealed that the process of ordering is triggered by attractive lateral capillary forces due to the overlap of the menisci formed around the particles. Two types of lateral capillary forces, flotation and immersion, can be distinguished, and a theory of these interactions is developed. Similar forces are operative between inclusions (proteins) incorporated in lipid membranes. We develop an appropriate model of a lipid bilayer, which is described as an elastic layer (the hydrocarbon chain region) sandwiched between two Gibbs dividing surfaces (the two headgroup regions). The range of the interaction between two cylindrical inclusions turns out to be of the order of several inclusion radii. The results, which are in qualitative agreement with the experimental observations, can be applied to the interpretation of membrane processes and mechanisms such as protein aggregation in lipid membranes. PMID- 9204125 TI - Significant role of electrostatic interactions for stabilization of protein assemblies. AB - Contribution of electrostatic interactions to stability of BPTI orthorhombic, pig insulin cubic crystals, and horse L ferritin crystals was evaluated with numerical calculation of Poisson-Boltzmann equation based on a dielectric model. The stability of a ferritin molecule (24-mer) composed of 24 subunits was also evaluated. It was found that the surface charge-charge interactions at separation distances (< 5 A) were insensitive to variations in the ionic strength, and thus stabilized assembled states of the proteins (i.e., crystalline state and oligomeric state). It was also revealed that the charge density and the packing of the protein crystals were largely responsible for the ionic strength dependence of the crystal stability. The stability of the 5PTI crystalline state with a high charge density drastically increased as the concentration of the solvent ions increased. In contrast, that of the insulin crystal with a low charge density and large solvent region was insensitive to changes in the ionic concentration. The electrostatic interaction between ferritin 24-mers was attributed to two salt bridges mediated by Cd ion. For the stability of the ferritin 24-mer, which is evolutionally designed, the electrostatic stabilization between the subunits was attributed to polar bonds such as buried salt bridges or hydrogen bonds, which occasionally yielded more than 5 kcal/mol and were numerous and very strong compared with the bonds between molecules in the 5PTI and 9INS crystals. By analyzing the atomic charge-charge interactions in detail, it was found that charge pairs separated by less than 3 A, such as hydrogen bonds, dominantly stabilize the assembled states, and that pairs 3 to 5 A apart were also important. The stability of the assembled states evaluated by the total EET was determined by the fine balance between the two competing contributions arising from the stabilizing atoms and the destabilizing atoms. Changes of the ASA and hydration free energy were also evaluated in accordance with the process of the subunit assembly. The change of hydration free energy, which was very large (i.e., approximately +100 kcal/mol/subunit) and unfavorable for the assembly, was proportional to the electrostatic hydration energy (i.e., Born energy change in the hydration process). Hydrophobic groups were likely to appear more frequently than hydrophilic groups at the interfaces. This study offers a method which can improve the stability of protein crystals by introducing polar or charged residues that are properly designed to form specific hydrogen bonds or salt bridges between neighboring protein molecules. This method is also applicable to crystallography, because it improves refinement of protein structures in crystals by taking the inter-protein interactions into account. PMID- 9204126 TI - Criticality found in a model for orientational ordering of protein arrays. AB - MC-PSRG analysis allowed us to reduce criticalities A, B, and C found in the poker chip model, respectively, to ones of the Ising universality, of the 3-state Potts universality, and of the KT-like phase. We note that not only the KT-like phase but also the Ising and 3-state Potts universality have been predicted to appear in the generalized 6-state clock model. We expect that the criticality inherent in actual protein array systems, whose Hamiltonians might be more complex than those of the poker chip model, can also be reduced to the criticality clarified with the aid of the naive models. However, we have to direct our attention to nonuniversal behavior like criticality C. Several two dimensional systems having the two-component order parameter exhibit nonuniversal critical behavior, whose critical exponents do not coincide with those for the XY model despite a coincidence in the number of order parameter components (17, 20, 25). PMID- 9204127 TI - Advances in S-layer nanotechnology and biomimetics. AB - Two-dimensional crystalline bacterial S-layers composed of identical protein or glycoprotein subunits turned out to be ideal materials for the development of biomimetic membranes and new approaches in molecular nanotechnology. These isoporous protein lattices have already been used as (i) structure for producing isoporous ultrafiltration membranes with very precisely defined molecular sieving properties, (ii) matrices for immobilizing monolayers of functional molecules, (iii) stabilizing structure for LB-films and liposomes, and (iv) patterning elements in molecular nanotechnology. PMID- 9204128 TI - Assembly process of 2D protein arrays in wetting films. AB - We were successful in developing a technique to form protein array developed directly on solid surfaces. This promising array formation revealed two new concepts of the ordering mechanism: the lifetime of a secondary minimum and secondary films. These concepts are not limited to our film formation technique. Any electrolytic thin film on a solid surfaces or any free film may contain a secondary film. If the secondary film can be retained in an electrolytic thin film for a sufficient period, a large single domain of small colloidal particles, such as proteins, fine metal particles, fine semi-conductive particles, etc., can be produced by an Alder-type transition. PMID- 9204129 TI - Two-dimensional crystals of apoferritin. AB - A simple 2D crystallization method using unfolded protein film as a supporting film of crystals was described, which allows modification of protein surfaces by injecting chemical reagents into the subphase after the crystal formation. As an example, glutaraldehyde was used to cross-link adjacent proteins and then stabilize protein crystals. The second layer of other proteins can also be formed on the apoferritin array using cross-linkers. The array of apoferritin is not only beneficial for electron crystallography but also for practical applications. For example, apoferritin produces a mineral core with a size which can be adjusted by the size to the cavity (i.e. 6 nm). Fabrication of such a small size of well defined fine particles is currently not easy using physical or chemical procedures. Using apoferritin, however, it is easy to produce uniform fine particles. If the core is designed to add interesting properties such as magnetism it is possible to make the highest class of magnetic film with ferritin 2D crystals. Basic researches toward practical applications of 2D protein crystal is now under way in various fields. The well defined size and function of protein molecules will benefit to many applications. The function and crystalline order can be designed by site-directed mutagenesis with the development of protein engineering. PMID- 9204130 TI - Molecular handling of photosynthetic proteins for molecular assembly construction. AB - Methods of constructing proteins were examined with special reference to the molecular assembly using photosynthetic RCs as membrane proteins. Molecular assemblies at the interfaces were studied by LB films, adsorption to the surface and reconstitution into liposomes and bilayer lipid membranes. The applications of biological specific ligands (recognition and binding), combinatorial chemical method, 2-D and 3-D order array assemblies and modification of protein molecules to make fusion proteins, as well as physical methods of manipulation of molecules by AFM tips and electric fields were reviewed. PMID- 9204131 TI - Supramolecular assembly using helical peptides. AB - We investigated supramolecular assemblies of various hydrophobic helical peptides. The assemblies were formed at the air/water interface or in aqueous medium. The hexadecapeptide, Boc-(Ala-Aib)s-OMe (BA16M), was reported to take alpha-helical structure by X-ray analysis. Several derivatives were prepared, which have the repeating sequence of Ala-Aib, Lys(Z)-Aib or Leu-Aib, or have the terminal chemically modified. CD spectra of the peptides indicated helical conformation in ethanol solution. The surface pressure-area isotherms of the peptide monolayers showed an inflection at the surface area corresponding to the cross section along the helix axis, and the monolayers were collapsed by further compression. All the helical peptides oriented their helix axis parallel to the air/water interface on the basis of the results of transmission IR spectra and RAS of the monolayers transferred onto substrates. A small mound was observed in the isotherm of BA16M and other derivatives, which was ascribed to the phase transition from the liquid state to the solid state. One mol% of FITC-labeled peptide was mixed into the monolayers to visualize the phase separation of the solid and liquid states at the surface pressure of the coexisting region. Various shapes of the dark domain were observed at the top of the mound in the isotherms by fluorescence microscopy. The helical peptides formed two-dimensional crystals at the air/water interface when they were compressed to the solid state. An amino terminated helical peptide, HA16B, was suspended in an aqueous medium by a sonication method and transparent dispersion was obtained. The dynamic light scattering measurement of the dispersion revealed the particle size of 75 nm with a narrow size distribution. The molecular assembly of the helical peptide in water was called "Peptosome", because it takes a vesicular structure. PMID- 9204133 TI - Electron crystallography of macromolecular periodic arrays on phospholipid monolayers. AB - Electron crystallography has the potential of yielding structural information equivalent to x-ray diffraction. The major difficulty has been preparing specimens with the required structural order and size for diffraction and imaging in the electron microscope. 2D crystallization on phospholipid monolayers is capable of fulfilling both of these requirements. Crystals can form as a result of specific interactions with a protein's ligand or an analog, suitably linked to a lipid tail; or on a surface of complementary head-group charge. With such choices, the availability of a suitable lipid is limited only by synthetic chemistry. Ultimately, it is the quality and regularity of the protein-protein interactions that determine the crystalline order, as it is with any protein crystal. In the case of streptavidin, the monolayer crystal diffracts beyond 2.5 A. A 3 A projection map reconstructed from electron diffraction amplitudes and phases from images shows density which can be interpreted as beta-sheets and clusters of side chains. It remains to be shown that the monolayer crystals are flat and diffract as well at high tilt angle as untilted. Technological issues such as charging must be resolved. With parallel advances in data collection and processing, electron crystallography of monolayer macromolecular crystals will eventually take its place beside x-ray crystallography and NMR as a routine and efficient structural technique. PMID- 9204132 TI - Assembly of protein structures on liposomes by non-specific and specific interactions. AB - We investigate different schemes for fabrication of nanometer sized assemblies that consist of a liposome core over which a shell of ferritin is attached. Three distinct interactions were used for this assembly: (i) Electrostatic attraction. The liposomes are charged by the presence of cationic surfactant (HTAB) and at an appropriate pH collect the ferritin molecules into a 2D-ordered ferritin shell. The protein shells can be fixed by glutaraldehyde. Next, the liposomes can be removed by solubilisation, leaving behind ordered ferritin clusters. (ii) Specific avidin-biotin or streptavidin-biotin binding. The ferritin molecules are conjugated to avidin or streptavidin and the liposomes incorporate biotinylated lipid. We found that the specific binding can be completely blocked by unfavourable electrostatic repulsion. To adjust the appropriate liposome charge we include cationic surfactant in the lipid layer. Thus, to accomplish the assembly process, we need to design and modify both the specific and non-specific colloid interactions in the system. The result is liposomes heavily coated with a strongly and specifically attached ferritin layer. (iii) Specific polysaccharide/lectin binding. The liposomes are first coated with a cholesterol anchored mannan layer. The ferritin molecules are conjugated with Con A that binds to the polysaccharide. A smooth and dense coating with ferritin is obtained (Fig. 3b). The acquired data can find application in the future fabrication of microstructured, multicomponent, or functionalised protein and liposome/ protein assemblies. PMID- 9204134 TI - The influence of protein and interfacial structure on the self-assembly of oriented protein arrays. AB - These results demonstrate the complexity of factors that impact the self-assembly of protein arrays via the specific binding to receptor-functionalized interfaces. Both the composition and colloidal properties of the protein and target membrane surfaces will affect the protein-surface interactions. However, different structural features control the interactions over different distance regimes and with different consequences. The long-range interactions that control the adsorption kinetics are sensitive not only to the charge on the target surface but also by the topological charge distribution on the protein exterior. Short range repulsive interactions rooted in both the protein topology and in the membrane structure, by contrast, can significantly alter both the rates and strengths binding. Consequently, the effective design of self-assembling protein arrays must consider not only the recognition interactions that drive such self organization, but also the details of the micro-environment and their impact on molecular recognition events. PMID- 9204135 TI - Single molecular functional assay of ferritin arrays. AB - In situ functional assay of each ferritin molecule in single-layer 2D arrays for horse spleen apoferritin and recombinant horse L- and human H-apoferritins was conducted by observing the iron-cores formed in the arrays by TEM. The study of the time-course, pH-dependence, and temperature-dependence of the function confirmed the iron-core formation to be due to the native function of apoferritins in array. Dark-field TEM imaging revealed that there was crystallinity in the cores in the array of recombinant human H-apoferritin. This iron-core formation was perfectly preserved in the array even after 3 months of storage at room temperature and low humidity. Moreover, about 50% of the function was found to remain in the array after it was exposed to 150 degrees C in vacuum for 1 hr. PMID- 9204136 TI - Novel biosensoric devices based on molecular protein hetero-multilayer films. AB - We have developed a novel concept for the modification of technical surfaces with molecularly well-organized layers of bioorganic components. A molecular construction set has been used to implement this concept which is based on molecularly stratified polyelectrolyte films as a structure decoupling protein layers from solid substrates. Utilizing this technology, one can start from a number of different substrates to obtain the same surface structures, on which protein hetero-multilayer films can be prepared to functionalize the interface for (potentially very different) purposes. We have demonstrated the viability of this concept by constructing a biosensor surface that was characterized by x-ray, spectroscopic, and immunological techniques. For the preparation of this functionalized interface, a cascade of biospecific recognition processes has been used, in which a layer of SpA was supplanted on a dense SA monolayer (binding stoichiometry, approximately 0.2 SpA/SA). On the SpA, IgG has been immobilized (binding stoichiometry, approximately 0.5 IgG/ SpA). The resultant biosensor displays extremely low unspecific background reactivity, high sensitivity, and good response linearity over more than 3 orders of magnitude in antigen concentration. PMID- 9204137 TI - Supramolecular architectures for the functionalization of solid surfaces. AB - Surface plasmon optical techniques are described as sensitive tools that allow for the on-line characterization of supramolecular biofunctional architectures at solid/solution interfaces. After a short introduction into the fundamentals of surface plasmon optics the observation of the build up of a functional bio interface by the self-assembly process of long chain thiolates at an Au surface is described. Criteria are developed for tailoring the SAM architectures optimized for maximum protein binding from solution by specific bio-recognition reactions. SPM is employed to image the selective binding of streptavidin to a functionalized SAM laterally patterned by UV-photolithographic techniques. PMID- 9204138 TI - Natural and artificial carbohydrate-glued protein aggregates. AB - Carbohydrate gluing (which may have a carbohydrate-lectin binding mechanism) was first recognized as a major contributor in the supramolecular assembly of annelid giant Hb from the marine-worm P. aibuhitensis. Although this assembly obviously also relies on protein-protein interactions, the authors tested the application of carbohydrate gluing in the assembly of a protein aggregate using a lectin and a carbohydrate-containing protein. The resultant aggregate was a mixture of the protein aggregate and the ingredient proteins. The significance of the method is that the assembly of the aggregate can be controlled by using a hapten sugar. This controllability, in conjunction with newly developing glyco-technology, has great potential for the construction of arbitrary protein molecules into a regular protein aggregate, thereby providing sophisticated functions. PMID- 9204139 TI - Apoptosis: an overview of the process and its relevance in disease. PMID- 9204140 TI - Genetics of apoptosis. PMID- 9204141 TI - Methods utilized in the study of apoptosis. PMID- 9204142 TI - In vitro systems for the study of apoptosis. PMID- 9204143 TI - The Fas pathway in apoptosis. PMID- 9204144 TI - Ceramide: a novel lipid mediator of apoptosis. PMID- 9204145 TI - Control of apoptosis by proteases. PMID- 9204146 TI - Death and dying in the immune system. PMID- 9204147 TI - Control of apoptosis by cytokines. PMID- 9204149 TI - Apoptosis in AIDS. PMID- 9204148 TI - Glucocorticoid-induced apoptosis. PMID- 9204150 TI - Virus-induced apoptosis. PMID- 9204151 TI - Apoptosis in neurodegenerative diseases. PMID- 9204152 TI - Apoptosis in the mammalian kidney: incidence, effectors, and molecular control in normal development and disease states. AB - In the preceding sections we have emphasized the current status of our knowledge concerning the involvement of apoptosis in normal and abnormal renal developmental processes, in control of the adult kidney size and capacity, in the development of renal disease states and in renal oncogenesis. At several points, we noted that studies of apoptosis in the kidney and in renal cells lag behind those in other organ systems. Even with the rudimentary knowledge now available, however, it is apparent that apoptosis is an extremely important process in the kidney. Recent observations lend credence to the view that continued study of this unique cell death process might enable the generation of novel and more effective therapies to treat renal diseases and renal malignancies. We wish to highlight several areas that require particular attention. First, the relationship between blood supply and apoptosis in the kidney requires further investigation. Benign human renal diseases are common in our population; and we now know that most of these diseases are associated with abnormal rates of apoptosis. Although the initiating agents for the various renal diseases vary, there is good reason to believe that much of the apoptosis that occurs in these adult diseases is the end result of reduced renal blood flow initiated by the causative agent. Cytokines or other extrinsic agents that can reduce the apoptotic loss of renal cells under these conditions hold theoretical promise in treating these diseases. Second, there is an urgent need to define the endocrine, paracrine, or autocrine roles of cytokines in normal renal physiology and in the pathogenesis of various renal syndromes. As indicated above, elaboration of fibrous extracellular material by fibroblasts in the tubulointerstitial regions of the kidney appears to be part of the final common pathway leading to end-stage renal disease. It is important to understand how the function of these fibroblasts is controlled. Conversely, apoptosis of glomerular or renal tubular cells also appears to play a role in the development of many of these syndromes. There is already experimental and clinical evidence showing that IGF-1 and hepatocyte growth factor therapies can be useful in renal diseases [57, 58]. It remains to be determined how much of the usefulness of these cytokines is related to their ability to suppress apoptosis as opposed to their ability to promote true growth. Finally, the analysis of apoptotic regulation during renal oncogenesis is critical. Maligant renal cell cancers are difficult to detect in adults before their metastases cause symptoms; and by this late stage renal tumors are almost invariably fatal. The ability of these tumors to regress spontaneously indicates that most apoptotic pathways are retained in these cells, yet their disappointing response to chemotherapy indicates that we have much to learn about how to trigger these pathways. Hopefully a better understanding of the control of these pathways will lead to improved therapy for this devastating group of neoplasms. PMID- 9204153 TI - Apoptosis in the heart. PMID- 9204154 TI - Apoptosis and the gastrointestinal system. PMID- 9204155 TI - Role of p53 in apoptosis. PMID- 9204156 TI - Chemotherapy-induced apoptosis. PMID- 9204158 TI - Role of Bcr-Abl kinase in resistance to apoptosis. PMID- 9204157 TI - Bcl-2 family proteins: strategies for overcoming chemoresistance in cancer. PMID- 9204159 TI - Apoptosis in hormone-responsive malignancies. PMID- 9204160 TI - Endoscopic facelift: two years' experience. AB - In the upcoming Twenty-first Century, we will find many surgical methods and devices that come to fulfill one of the main objectives of the aesthetic plastic surgery: to reduce scars, especially in facial surgery. Endoscopy is one of those methods. In my experience of the last two years, I have used this technique, sometimes combined with CO2 laser to partially remove glabellar muscles and the platysma fibers of the middle part of the neck. This work shows the results from 160 patients undergoing endoscopic forehead lift and neck contouring, using specially designed instruments. The results are highly significant and satisfactory. PMID- 9204161 TI - Chest wall implants: their use for pectus excavatum, pectoralis muscle tears, Poland's syndrome, and muscular insufficiency. AB - Solid customized and prefabricated silicone implants have been used by the author for 15 years in a wide range of chest wall deformities. Chest wall implants are often used in males seeking to augment a muscularly deficient or underdeveloped chest; however, their greatest use has come in a variety of deformities both congenital and acquired, such as pectus excavatum, Poland's Syndrome, and pectoralis muscle tears. The implants can be either customized using a moulage technique or are prefabricated, manufactured implants which can be modified on the operating table to repair the contour deformity. The immediate postoperative problem of seroma and subcutaneous implant "show" has been minimized by careful planning, gentle technique, deep insertion, improved patient positioning on the operating room table, and the use of oral anti-inflammatory medications. The long term results of these implants seem very satisfactory. The patients are usually physically active, and the implants show no long-term sequelae such as seroma, infection, displacement, or rupture. PMID- 9204162 TI - Aesthetic breast surgery with inverted-T scar placed above the inframammary sulcus. AB - This article describes the inverted-T incision technique with the scar placed above the inframammary sulcus for cases of pexy, breast reduction, and augmentation-reduction mammaplasty. This technique preserves the inframammary fold as an important factor in natural breast suspension; the breast mound is easily shaped independent of the skin tension. The gland- and skin sutures are placed separately and independently. This technique has been used on 380 patients in the last 13 years. PMID- 9204163 TI - Pain reduction in breast augmentation using methocarbamol. AB - Submuscular placement of breast implants produces significant postoperative pain and discomfort. The standard use of narcotics alone does not optimize pain reduction. Methocarbamol was used intraoperatively and postoperatively in 62 patients undergoing manipulation of the pectoralis major associated with breast implant surgery. Significant pain relief was achieved. PMID- 9204164 TI - Cervicofacial rhytidectomy: the superficial plane. AB - During the past 20 years, numerous techniques have been described to improve the results of facelifting as well as the duration of its rejuvenating effect. These increasingly aggressive techniques have increased the operating time, the potential morbidity of the operation, and the duration of convalescence. No one can ensure that this evolution in sophistication of rhytidectomy techniques give longer lasting results. Therefore, a simple technique applied under the appropriate indications gives good and predictable results. PMID- 9204165 TI - Neck rhytidectomy: aesthetic treatment variations. AB - The aesthetic treatment of the cervical region had its real importance in neck rhytidectomy, after the publications of, among others, Padgett, Cardoso de Castro, Millard, Illouz. The author makes an analysis of various cervical pathologies, their treatment with lipoaspiration, lipectomy, and the connection of the platysma muscle (isolated and in conjunction). Pre- and postoperative aspects of cases of diverse neck deformities, and surgical procedures are discussed. Surgeries were done under sedation or local anesthesia as the case dictated. The importance of case-specific treatment for each patient is discussed. The author demonstrated one complication in a patient who underwent neck rhytidectomy before the therapeutic possibilities of today were developed. PMID- 9204166 TI - Is there any real advantage in the double-lumen cannula? AB - The author demonstrates one of liposuction's great problems, what he calls the "contra-vacuum," which forms inside the cannula while it is inserted under the skin. To keep liposuction effective, an ascending pressure is needed while the holes of the cannula are deep inside the tissues. The results are higher pressure, more tissue trauma, less effectiveness. To prevent these problems, the author developed a double-lumen cannula in 1983; because of a secondary tube outside it, there is no contra-vacuum formation. He presents here his experience in large liposuctions with a new cannula model, thinner than the first one, but also very effective. PMID- 9204167 TI - Alar lateral crus in nasal tip surgery. AB - The authors consider the morphology of alar lateral crus as a means of classification. Every variety of lateral crus has a peculiar property of shape and relationship with other elements that determines the course of therapy. Diagrams for explication, descriptions of surgical techniques, and clinical results are included here. PMID- 9204168 TI - Needle dermabrasion. AB - In this article we describe a technique of needle dermabrasion (tattoo without pigment) used to improve achromic, hypertrophic, and unsightly scars. It is simple, safe (no complications), and it gives us consistently good results. PMID- 9204169 TI - Treatment of the before and after images in aesthetic surgery. AB - The author presents a new system of treatment using instant photographs for imaging body and facial recontouring before and after aesthetic surgery. The surgeon draws on photocopies of the photograph, blotting out in black to get a sense of reduction and adding with a white corrector to get a sense of enlargement. PMID- 9204171 TI - Ethnic profile of patients undergoing aesthetic rhinoplasty in Stockholm. AB - During 1985-1995 we performed 640 rhinoplasties in 578 patients. Five hundred eighteen of them were inhabitants of the Province of Stockholm, with a population of 1,708,502. The patients from the Stockholm area were analyzed and divided into subgroups depending on their ethnic origin. It was found that 272 (52%) of them were of Nordic descent, while 248 (48%) were born in and immigrated from non Scandinavian countries. Among the latter, the largest group were 166 people of Middle Eastern extraction, who generally strived to reduce the size of their noses to the size similar to the average nose of the native Swedes. Middle Easterners were 17 times more prone to undergo aesthetic rhinoplasty than the ethnic Swedes (p < 0.001), whereas immigrants from the other Scandinavian countries had the same rhinoplasty frequency pattern as the natives. In the Slavic group females outnumbered males by the ratio 17:1. The large prevalence of patients of foreign extraction desiring alteration of their noses may reflect the assimilation difficulties and low tolerance of the society in accepting people with a foreign look or name, both in the private sector and in the job market. Psychological aspects of decision making by patients and medico-ethical aspects of decision making by surgeons are discussed. PMID- 9204170 TI - Deepithelialized vertical rectus abdominis flap in calf augmentation. AB - This is the first report of a case of aesthetic calf augmentation with microvascular transferred autologous material. The technique regarding to calf augmentation using silicon prostheses is described and the advantages and disadvantages of this method are discussed. This should incite discussion on the use of free-flap technique in aesthetic operations. PMID- 9204172 TI - Nasal speculum with positionally autoswitched and battery-operated fiber optic illumination. AB - We modified a Cottle-type nasal speculum with fiber optics to a positionally autoswitched and battery operated device. By avoiding the use of a fiberoptic cable, our speculum proved to be a practical and effective tool for intranasal illumination. PMID- 9204173 TI - Benefits of the rotating cannula in the treatment of lipodystrophies. AB - The combination of traditional suction and the use of a cannula which can achieve up to 400 rotations per minute can enable the practitioner to treat lipodystrophies at an angle of 360 degrees. The torque limitation device puts an immediate end to rotations if the cannula meets a resistance, thereby eliminating any risk of lesions to non-fatty tissues. The use of this device over a one year period has proved its efficiency in the treatment of major lipodystrophies and certain areas, which are usually considered difficult because of their density. It is of limited interest, however, in the correction of moderate, classical lipodystrophies. PMID- 9204174 TI - Morpho-histological analysis of abdominal skin as related to liposuction. AB - The analysis of the abdominal skin on cadavers has made it possible for the authors to evaluate the skin's regional characteristics, taking into consideration the age group to be lipoaspirated regardless of race or sex. In this way, norms are established which in the future may be used to comparatively analyze results related to the cutaneous retraction after plastic surgery lipoaspiration. PMID- 9204175 TI - Combined aesthetic and functional treatment of microtia. AB - The reconstruction of the ear pavilion includes at least two stages, usually three, and in many cases more. Complete reconstruction must begin with the plastic surgeon, who sculpts the cartilaginous skeleton, assembles parts of three ribs, and inserts the finished framework under the expanded skin of the mastoid area. During the second stage, the otologist intervenes, performing the cophosurgery, carving an external auditory canal, and completing the ossicular chain in the middle ear. The plastic surgeon harvests the skin for lining the external auditory canal and for the retroauricular fold and forms the lobule with a part of the microtic vestige. The third stage is dedicated to refinements. Cophosurgery may also be performed during the third stage or in an interval between two stages of pavilion construction. Twenty-one cases are discussed. PMID- 9204176 TI - Importance of fat conservation in lower blepharoplasty. AB - The authors describes her experience in over 200 cases operated on from 1993 to 1996 with the lower blepharoplasty with arcus marginalis release and fat conservation as described by Hamra [Plast Reconstr Surg 96:354, 1995; Clin plast Surg 23:17, 1996]. The overall satisfaction with the somewhat modified technique led the author to abandon completely the conventional type of fat resection. The technique has been used in different clinical situations. The anatomical basis, the technique, and the complications are discussed. PMID- 9204177 TI - Breast reconstruction by expansion and advancement of the upper abdominal flap. AB - Inspired by successful reconstruction obtained using the Lewis-Ryan lower thoracic advancement flap to rebuild missing breast, we have adapted that extremely simple technique to prior serial expansions, in order to create more natural mounds, better defined submammary folds, and when possible, some grade of ptosis, without additional, new scarring. The procedure is introduced and compared to other such flaps as the TRAM and the latissimus dorsii. In our series, 30 patients were evaluated according to the quality of the final results, and the most frequent complication are pointed out and discussed. PMID- 9204178 TI - Wineglass pattern for vertical mammaplasty. AB - The ideal reduction mammaplasty technique should create a pleasing breast shape with minimal scarring. The long and conspicuous scar associated with the classic inverted "T" pattern mammaplasty techniques are not acceptable for many patients. Periareolar mammaplasty techniques cause less scarring, but they have major disadvantages such as scar widening, areolar distortion, and insufficient breast projection. We used a new pattern for vertical mammaplasty to overcome the insufficient breast projection caused by the round block technique and applied it to 51 patients during the last 3 years. This method results in a single vertical scar and a periareolar scar, allows sufficient volume reduction, and provides good breast shape and projection; the results are durable. This procedure is safe, causes few complications, and is easy to learn and perform. PMID- 9204179 TI - Mammary hypertrophy: a personal approach. PMID- 9204180 TI - An improved method of removing polyurethane foam-covered gel prostheses. AB - One of the advantages of polyurethane foam-covered prostheses has been that in the first 5 to 10 years after their use, the amount of capsular contraction was found much less than when similar "slick" prostheses were used. Another advance was their fixation to the surrounding tissue thus giving a more natural appearance and movement with the muscles when the arms were moved in any direction. The formation of a thick capsule also acted as a protection against gel granuloma due to rupture of the prosthesis and has been thought to be a factor in the lower capsule contraction rate. The greatest disadvantage has been that its removal was extremely difficult and this has continued up until the technique described in this paper has been introduced. PMID- 9204181 TI - Correction of inverted nipple with modified star flap technique. AB - The authors describe the application of the modified star flap technique for correction of inverted nipples. This technique allows easy identification and dissection of the lactiferous ducts under a clear direct view without trans nipple incision. The secure deep dermal buried sutures around the nipple base in three directions maintain the newly everted nipple. The blood supply to the nipple flap is reliable through the dermal and subdermal plexus from the remaining non-incised site of the nipple and areola region. PMID- 9204182 TI - Injection of all-trans retinoic acid for treatment of thin wrinkles. AB - The authors propose a treatment to improve skin texture and to decrease thin wrinkles and creases. The treatment is based on the use of 0.1% all-trans retinoic acid intradermic injections (with biopresence of 0.02%) combined with topic cream, immediately followed by 340 Nm blue light skin exposure. These procedures determine the retinoic binding protein stabilization that provides the acid intracellular penetration with its subsequent effects. An average of 10 sessions, once a week was required. PMID- 9204183 TI - Yucatan pig: an optimal hairless model for a true random cutaneous flap. AB - Porcine models have been used extensively for skin flap research because of the established similarity between the cutaneous blood supply of the swine and humans. The Yucatan minipig provides an excellent model for researching the properties of random cutaneous flaps, offering several advantages over other breeds of swine. In this study, a total of 67 random cutaneous dorsal flank flaps measuring 4 x 14 cm were raised on nine Yucatan minipigs. The mean survival length (10.03 +/- 1.60 cm) of the nondelayed flaps was greater than others reported in the literature. The well-defined plane between the subcutaneous tissue and the panniculus carnosus facilitated flap elevation consistently above the level of the panniculus carnosus thereby ensuring the creation of a true random cutaneous flap. Furthermore, the hairless nature of the skin, particularly beneficial in studying chemical peels, permits easy visualization and monitoring of any external skin changes. These advantages make the Yucatan minipig a more desirable alternative to other breeds of swine for use in skin flap research. PMID- 9204184 TI - An alternative treatment for the split skin-graft donor site. AB - There are a variety of methods employed in the postoperative management of the partial thickness donor site created during harvest of a split thickness skin graft. Each technique may be associated with potential complications of fluid loss, excessive pain, prolonged period for healing and delayed mobility, hypertrophic scarring, undesirable pigment aesthetics, and thin skin poorly resistant to everyday trauma. Thompson, and Converse and Robb-Smith have previously shown improved donor site outcome with the application of thin split skin grafts. Based on these studies, we present a technique that involves 1.5:1 meshing of a split skin graft and dividing it into equal halves so that one half is used to cover the defect and the other half is immediately returned to the donor site. Patients who are elderly, debilitated, or who have thin, poor-quality skin can expect less discomfort, reduction of fluid loss, improved durability and elasticity, and lower incidence of hypertrophic scarring with the proposed donor site regrafting. PMID- 9204185 TI - Growth hormone- and growth-hormone-releasing hormone-producing tumors. PMID- 9204186 TI - Adrenocorticotrophic hormone-dependent Cushing's syndrome. AB - Excess endogenous glucocorticoid production, whether ACTH dependent or ACTH independent, results in the classic clinical and biochemical picture of Cushing's syndrome. The diagnosis requires the demonstration of an increased cortisol secretion rate, best achieved by using the 24-hour UFC corrected for body surface area as an index. In mild cases, distinction from the hypercortisolism of pseudo Cushing states may be difficult. A dexamethasone/o CRH test or close monitoring of the patient for a few months may be helpful. A discrete pituitary lesion on imaging and a standard oCRH test with results consistent with such a lesion are sufficient to proceed to trans-sphenoidal surgery. If no visible pituitary adenoma is present or if the oCRH test is equivocal, bilateral simultaneous inferior petrosal sinus sampling with oCRH administration is necessary to distinguish between a pituitary and an ectopic source. Surgical ablation is the treatment of choice for all types of Cushing's syndrome. In the 5% of cases with Cushing's disease in whom trans-sphenoidal surgery fails and in the 5% of cases in whom the disease recurs, repeat trans-sphenoidal surgery or radiation therapy in association with mitotane treatment may be pursued. Bilateral adrenalectomy effectively cures hypercortisolism if resection of the ACTH-secreting tumor is unsuccessful and radiation/medical therapy fails. PMID- 9204187 TI - Prolactinomas. PMID- 9204188 TI - Gonadotroph and other clinically nonfunctioning pituitary adenomas. PMID- 9204189 TI - Approach to the incidentally discovered pituitary mass. AB - Incidental pituitary adenomas are being found commonly with our improved neuroradiologic imaging procedures. Screening for hormone oversecretion by these tumors appears to be warranted. For patients with macroadenomas, patients should also be screened for hypopituitarism. In the absence of visual-field abnormalities or hypothalamic/stalk compression, it may be appropriate to observe such patients carefully with repeated MRI scans. A limited amount of data suggest that significant tumor enlargement will occur in < 5% of patients with microadenomas [8,11]. However, all macroadenomas must start out as microadenomas, and so periodic follow-up is indicated to assess for this possibility. Macroadenomas, by their very existence at the time of detection, have already indicated a propensity for growth. Over the limited period of follow-up in the two series reported, significant growth occurred in just over one quarter of the patients with macroadenomas [8,11]. Hemorrhage into such tumors is uncommon, but anticoagulation may predispose to this complication. When there is no evidence of visual-field deficits, an attempt at medical therapy with a dopamine agonist or octreotide is reasonable, realizing that only about 10% of such patients will respond with a decrease in tumor size. Surgery is indicated if there is evidence of tumor enlargement, especially when such growth is accompanied by compression of the optic chiasm, cavernous sinus invasion, or the development of pituitary hormone deficiencies. PMID- 9204190 TI - Follicular cell-derived thyroid carcinomas. PMID- 9204191 TI - Radiation-induced endocrine tumors. PMID- 9204192 TI - Diagnosis, natural history, and treatment of primary hyperparathyroidism. PMID- 9204193 TI - Parathyroid carcinoma. PMID- 9204194 TI - Parathyroid hormone-related peptide and hypercalcemia of malignancy. PMID- 9204195 TI - Neoplasms of the adrenal cortex. Clinical and basic aspects. PMID- 9204196 TI - Pheochromocytoma and primary aldosteronism. PMID- 9204197 TI - Incidentally discovered adrenal masses. PMID- 9204198 TI - Gastrin-producing tumors. PMID- 9204199 TI - Insulinoma and other islet-cell tumors. PMID- 9204200 TI - Persistent hyperinsulinemic hypoglycemia of infancy. PMID- 9204201 TI - Somatostatin analogs and receptors. Diagnostic and therapeutic applications. PMID- 9204202 TI - Multiple endocrine neoplasia type I. Clinical genetics and diagnosis. PMID- 9204203 TI - Multiple endocrine neoplasia type I. Surgical therapy. PMID- 9204204 TI - Multiple endocrine neoplasia type II and familial medullary thyroid carcinoma. Impact of genetic screening on management. AB - The identification of ret protooncogene mutations in MEN-II and Hirschsprung disease has not only improved the clinical management of these genetic conditions but has also provided important information regarding mechanisms of transformation and neural crest development. An indication of how neural-crest cells migrate during embryonic life and the key processes involved in their differentiation now seems within reach. The continued pace of scientific discovery suggests that our understanding of and ability to prevent or treat hereditary and sporadic forms of MTC will continue to improve. PMID- 9204205 TI - Diagnostic performance of red cell sorbitol assay in a Nigerian teaching hospital. AB - The analytical linearity, detection limit, precision and accuracy, as well as diagnostic sensitivity, specificity and predictive values of red cell sorbitol (RCS) assay were evaluated using the fluorimetric enzymatic technique with the available laboratory facilities in a Nigerian Teaching Hospital. The assay was linear up to 40.0 nmol/ml. The detection limit was 0.55 nmol/ml. The within-assay coefficients of variation (CV) from low to high concentrations were in the range of 2.82% to 5.48% while the corresponding values for the between-assay variation were 3.60% to 6.05%. The mean recovery was 90.4%. High concentrations of some common blood constituents and drugs did not cause significant interference. An evaluation of the diagnostic sensitivity and specificity of RCS in 102 out patients revealed a sensitivity of 94.4%, specificity of 98.8% and efficiency of 98.0%. The predictive value of a positive test was 94.4% while that of a negative test was 98.8%. The mean value of red cell sorbitol in patients confirmed to have diabetes mellitus (23.1 +/- 1.13 SE nmol/g Hb) was significantly higher (P < 0.001) than the corresponding value (11.0 +/- 0.0.27 SE nmol/g Hb) for those without diabetes and for control subject. A similar observation was also made for plasma glucose and fructosamine. These observations demonstrate that RCS assay could be adopted in a third world laboratory for clinical applications. PMID- 9204206 TI - Early detection of successful coronary reperfusion based on serum concentration of human heart-type cytoplasmic fatty acid-binding protein. AB - Both human heart-type cytoplasmic fatty acid-binding protein (H-FABPc) and myoglobin are low molecular weight proteins that are abundant in the cytoplasm of myocardial cells. Unlike myoglobin, H-FABPc content in the skeletal muscle is less than in cardiac muscle. To investigate the usefulness of the serum concentration of H-FABPc in the early detection of successful coronary reperfusion, we measured serum concentrations of H-FABPc and myoglobin in 45 patients with acute myocardial infarction treated with intracoronary thrombolysis or direct percutaneous transluminal coronary angioplasty. Coronary angiography was performed every 5 min for reperfusion therapy to identify the onset of reperfusion. Reperfusion, defined as a TIMI grade 2 or 3, was achieved within 60 min of the initiation of reperfusion therapy in 30 patients (the reperfused group), but was not achieved in 15 patients (the non-reperfused group). Blood samples were obtained before initiation of treatment and 15, 30 and 60 min after initiation of treatment in the non-reperfused group. In the reperfused group, samples were obtained before reperfusion and 15, 30 and 60 min after reperfusion. The H-FABPc ratio (the ratio of value after to value before the initiation of treatment or reperfusion) increased sharply after the onset of reperfusion, peaking at 41 +/- 18 min, and decreased rapidly thereafter. The predictive accuracy of an H-FABPc ratio of > 1.8 for the detection of reperfusion within 60 min of the initiation of treatment was 93% at 15 min after reperfusion, 98% at 30 min, and 100% at 60 min. Similar rates of predictive accuracy were observed for a myoglobin ratio > 2.4. The H-FABPc ratio detected successful reperfusion as early as 15 min after the onset of reperfusion and was highly accurate in detecting reperfusion within 60 min of the onset of reperfusion. The predictive accuracy of the H-FABPc ratio was similar to that of the myoglobin ratio for the early detection of successful coronary reperfusion. PMID- 9204207 TI - Changes in plasma alpha 1-acid glycoprotein and albumin concentrations during late pregnancy in rhesus monkeys. AB - Plasma protein concentrations are an important determinant of maternal-fetal drug transfer in the perinatal period. Plasma concentrations of alpha 1-acid glycoprotein (AAGP) and albumin were measured in pregnant rhesus monkeys using radial immunodiffusion assays at weekly intervals during the third trimester (n = 12), at 3-day intervals in the 2 weeks prior to full term (n = 10), and at 0, 1, 12, and 60 h after birth (n = 4). AAGP concentrations increased significantly, and albumin concentrations decreased significantly, from weeks 13 to 21 gestation as well as from day 150 gestation to birth. No changes were seen in the postpartum period. AAGP and albumin concentrations were highly negatively correlated from day 150 gestation to birth. The data suggest that AAGP and albumin concentrations are jointly but inversely regulated by physiological or hormonal conditions that precede delivery. PMID- 9204208 TI - The effects of homocysteine and copper ions on the concentration and redox status of thiols in cell line cultures. AB - Different fractions (reduced and total) of thiols (homocysteine, cysteine and glutathione) were determined in HeLa cell cultures with and without addition of copper ions and/or homocysteine. In cell cultures without any addition the concentration of all intracellular thiols increased between 1 and 24 h of culture. Glutathione had the highest, whereas homocysteine showed the lowest, proportion of the reduced form. In the medium, there was a decrease of total cysteine during the incubation, but the amount of extracellular reduced cysteine increased. Both homocysteine and glutathione were released into the medium. The amount of exported homocysteine during the incubation exceeded several-fold the intracellular amount. There were no signs of cell toxicity induced by the high amounts of extracellularly added homocysteine (2000 mumol/l) in HeLa cell cultures, except a slight decrease in the concentration of intracellular glutathione. After the addition of copper ions (500 mumol/l) there was a retarded cell growth, decreased intracellular concentration of glutathione, increased release of glutathione into the medium and a lower proportion of all intra- and extracellular reduced thiols. After the addition of both copper ions and homocysteine to HeLa cell cultures, similar changes as with the addition of copper ions were noted except that the cell growth was still more retarded and that a very high level of intracellular homocysteine was noted at 1 h of incubation. N-acetylcysteine lowered, in these experiments, the intracellular accumulation of homocysteine and restored, to some extent, the cell growth. In an endothelial cell line even the presence of 500 mumol/l of homocysteine and 50 mumol/l of copper ions inhibited the cell growth and decreased the cellular level of glutathione. Whilst the levels of homocysteine in our short-time cell culture experiments are higher than the mild hyperhomocysteinemia thought to be atherogenic in humans (20-30 mumol/l), it is conceivable that over a longer time course (several decades) these lower levels of homocysteine in the presence of copper ions could be sufficient to induce cellular effects similar to those found in the present study, eventually leading to atherosclerosis. PMID- 9204209 TI - Lipoprotein (a) serum levels in patients with hepatocarcinoma. AB - Lipoprotein (a) [Lp(a)] is synthesised by liver cells, and patients with liver cirrhosis (LC) show low serum levels of Lp(a) associated with the degree of liver failure. On the contrary, increased serum levels of Lp(a) have been reported in patients with cancer. In this report, the behaviour of Lp(a) serum levels in patients with hepatocarcinoma (HC), a complication of LC, has been evaluated with the aim to study whether HC cells were able to cause an increase of serum concentrations of this lipoprotein when impaired liver protein synthesis is present. We selected eighteen patients affected by LC + HC, eighteen patients matched for sex, age and degree of liver failure with LC only, and eighteen patients with other cancer types. A significant increase of serum levels of Lp(a) was observed in patients affected by LC + HC or other cancer types compared with healthy subjects. Forty-four percent of LC + HC patients showed Lp(a) values more than 70.4 Units/dl, i.e., the upper limit of values observed in patients with LC only. Lp(a) serum concentrations were significantly associated with serum albumin both in LC and in LC + HC but not in other cancer-type patients. Thus, comparing patients with similar serum albumin concentrations, Lp(a) serum levels were significantly higher in patients with LC + HC than in patients with only LC and quite similar to those observed in patients with other cancer types. In conclusion, HC cells, in vivo, seem able to produce a greater amount of Lp(a) despite the reduced liver protein synthesis typical of LC. PMID- 9204211 TI - The evidence of oxidative stress in cardiac surgery and septic patients: a comparative study. AB - In present study, lipid peroxidation products (thiobarbituric acid reactive substances (TBARS) and diene conjugates (DC)) and markers of blood antioxidant status (serum antioxidative capacity (AOC) and red blood cells glutathione (RBC GSH)) were measured to compare the extent of oxidative stress in 12 cardiac surgery and 10 septic general surgery patients. In heart surgery, arterial TBARS were significantly increased 15 min after the start and 15 min after cessation of cardiopulmonary bypass, while AOC at these times was decreased. Eighteen hours after surgery all parameters, except antioxidative capacity, had returned to preoperative levels. In septic patients, the preoperative level of lipid peroxidation was significantly higher and antioxidative capacity lower than in heart surgery patients. Surgery had no influence on oxidative stress markers in this group of patients. Increase in lipid peroxidation and reduction in blood antioxidant capacity, induced either by sepsis or cardiopulmonary bypass, were of comparable extent. PMID- 9204210 TI - Model SV40-transformed fibroblast lines for metabolic studies of human prosaposin and acid ceramidase deficiencies. AB - Skin fibroblasts from patients with Farber disease (acid ceramidase deficiency) and from two siblings of the only known family affected with prosaposin deficiency were transformed by transfection with a plasmid carrying the SV40 large T antigen. The prosaposin-deficient transformed cell lines conserved their original metabolic defects, and in particular they were free of detectable immunoreactivity when using anti-saposin B and anti-saposin C antisera. Ultrastructurally, the cells contained heterogeneous lysosomal storage products. As found for their parental cell lines, the SV40-transformed fibroblasts exhibited deficient in vitro activities of lysosomal ceramidase and beta galactosylceramidase, but a normal activity of acid sphingomyelinase. As observed for SV40-transformed fibroblasts from Farber disease, degradation of radioactive glucosylceramide or low density lipoprotein-associated radiolabelled sphingomyelin by the prosaposin-deficient cells in situ showed a clear impairment in the turnover of lysosomal ceramide. Ceramide storage in prosaposin-deficient cells was also demonstrated by ceramide mass determination. In contrast to acid ceramidase deficient cells, both the accumulation of ceramide and the reduced in vitro activity of acid ceramidase in cells from prosaposin deficiency could be corrected by addition of purified saposin D. The data confirm that prosaposin is required for lysosomal ceramide degradation, but not for sphingomyelin turnover. The SV40-transformed fibroblasts will be useful for pathophysiological studies on human prosaposin deficiency. PMID- 9204212 TI - Effect of pirimiphos-methyl on proteolytic enzyme activities in rat heart, kidney, brain and liver tissues in vivo. AB - To elucidate whether pesticide toxicity in higher animals involves pesticide induced dysfunction of the intracellular protein catabolic process, we have determined the effect in vivo of the organophosphate insecticide pirimiphos methyl on the activities of representative protein catabolising cytoplasmic and lysosomal proteases (responsible for the various stages of the protein degradation cascade and essential for normal cell functioning) in heart, kidney, brain and liver target tissues in the rat. In liver tissue (the major site of pesticide metabolism), the activities of all of the cytoplasmic proteases investigated (alanyl-, arginyl-, leucyl aminopeptidases, dipeptidyl aminopeptidase IV, tripeptidyl aminopeptidase, proline endopeptidase) were significantly inhibited (by 20-40% of control activity) following administration of 10 mg pirimiphos-methyl/kg bodyweight, whereas of the lysosomal proteases investigated, only the activities of dipeptidyl aminopeptidase I and cathepsin D were significantly reduced (by 15-20% of control activity). In contrast, there was no insecticide-induced inhibition of protease activities in heart, kidney or brain tissues; some lysosomal enzymes (dipeptidyl aminopeptidase I, cathepsins L and D) showed significantly increased activities in these tissues (the reason for which remains to be determined). We conclude that the effect of pirimiphos-methyl on proteolytic enzyme activities differs in different target tissues, and that pirimiphos-methyl induced inhibition of proteases in liver tissue may represent a previously unrecognised toxicity hazard in higher animals. PMID- 9204213 TI - Prognostic significance of c-erbB-2/neu amplification and epidermal growth factor receptor (EGFR) in primary breast cancer and their relation to estradiol receptor (ER) status. AB - The aim of this study is to evaluate the prognostic significance of c-erbB-2/neu amplification and epidermal growth factor receptor (EGFR) expression in primary breast cancer (BC) and their prognostic implications when combined with estradiol receptor (ER) status. In this work, 825 BCs were studied. Neu amplification was evaluated by dot-blot and EGFR expression was evaluated by ligand binding assay using I125-EGF. Neu, EGFR, estradiol and progesterone receptors (ER and PR) had a marked influence on disease free survival (DFS) in univariate analysis. In node negative (NO) cases only neu was associated with short DFS (p = 0.005). However, in node-positive (N+) cases both EGFR (p = 0.005) and neu (p = 0.002) influenced DFS. None of the biological markers were significant predictors for overall survival (OS) in NO/BC. On the contrary, in N+/BC, EGFR + (p = 0.003) was associated with short OS. The EGFR + /neu/phenotype represented a sub-group with an even worse prognosis with respect to DFS (p = 0.0034) as well as EGFR + /ER tumors (p = 0.005). Moreover, neu + /ER-patients also had a high probability of relapse (p = 0.0000) and death (p = 0.006). C-erbB-2/neu, EGFR, histological grade, pN, pT and ER were subjected to a Cox multivariate regression analysis: neu was the most important parameter in predicting recurrence, and EGFR was a significant predictor for OS. PMID- 9204214 TI - An algorithmic approach using kappa/lambda ratios to improve the diagnostic accuracy of urine protein electrophoresis and to reduce the volume required for immunoelectrophoresis. AB - The most sensitive routine method for identifying urinary monoclonal immunoglobulin kappa and lambda light chains, called Bence Jones proteins (BJPs), in clinical laboratories is immunofixation electrophoresis (IFE), but this procedure is time-consuming and expensive. As a result, many laboratories screen for paraproteins with urine protein electrophoresis (UPE), which is insensitive when low concentrations of BJP are present and is difficult to interpret with severe proteinuria. The purpose of this study was to determine whether kappa/lambda ratios can be used in conjunction with UPE to improve diagnostic reliability in identifying paraproteins, and decrease the need for IFE on all samples. Urine specimens from 243 patients were examined by UPE and kappa/lambda ratios, and compared with IFE. Due to poor analytical sensitivity, the urinary kappa or lambda concentrations could not be determined in many cases. As a result, many specimens showed kappa/lambda ratios that were indeterminate. Nevertheless, when both urinary kappa and lambda concentrations were undetectable, a BJP could be ruled out. A urinary kappa/lambda ratio between 0.75 3 also ruled out a BJP. The use of kappa/lambda ratios, in conjunction with UPE, resulted in a 52% decrease in the volume of IFE during the course of this study, with 100% sensitivity for detecting BJP. PMID- 9204215 TI - Anti-galectin-1 autoantibodies in serum of patients with neurological diseases. AB - The presence of autoantibodies to human brain galectin-1 was investigated in serum from patients with multiple sclerosis, patients with or without evidence of other neurological disorders, and healthy controls, using an ELISA on purified brain galectin-1. Levels of autoantibodies to galectin-1 were significantly higher in patients than in healthy controls. Comparison of levels of anti galectin-1 and anti-idiotypic antibodies mimicking human brain galectin-1 (L-IgG) showed that the highest levels of autoantibodies were present in patients with low levels of L-IgG. This finding can be explained by hypothesizing that the concentration of autoantibodies to galectin-1 is possibly associated with impairment of the regulation of the immune system. PMID- 9204216 TI - Skeletal muscle troponin I release and magnetic resonance imaging signal intensity changes after eccentric exercise-induced skeletal muscle injury. PMID- 9204218 TI - Reference ranges for total homocysteine in children. PMID- 9204217 TI - Determination of extracellular matrix degradation by free radicals using viscosity measurement of hyaluronan. PMID- 9204219 TI - Touchdown PCR: a rapid method to genotype Helicobacter pylori infection. PMID- 9204220 TI - Comparing compliance patterns between randomized treatments. AB - When two equally efficacious drugs enter the market, the one with the better compliance is likely to be more widely used. Special management of the delivery may produce increased compliance. In this paper we analyze a trial of a single drug dosing prescription with patients randomized to either daily self monitoring of the outcome (blood pressure) or not. The study used Medication Event Monitoring Systems (MEMS) to record each exact time and date when a patient opened the pill container. No established method is available for comparing these high-dimensional compliance patterns between groups. This paper investigates several summary measures that highlight different dimensions of the pattern and the drug context in which they may be meaningful. Further, we examine conditional and marginal models that enable comparisons of the full pattern of daily dosing indicators for subjects between the groups. We found no simple difference in average compliance levels, but we found an interesting interaction between treatment and time: similar compliance existed initially among patients in both randomized groups, with a stronger decline over time for patients who did not monitor their blood pressure. We discuss how a balance between simplicity of interpretation and efficiency of data use may be sought in this case. PMID- 9204221 TI - Some statistical methods for multiple endpoints in clinical trials. AB - This paper summarizes, defines, and discusses multiple endpoints comparison procedures, concepts, and methodologies for applications to clinical trials. We address the more widely used methods of alpha-level, p-value, and critical value adjustments. We examine global assessment measures such as O'Brien's test and Simes' procedure and contrast them with the alpha-adjustment procedures of Bonferroni and Holm. We propose a global assessment procedure based on categorization of the individual endpoints to form an overall composite endpoint. Additionally, we discuss a new weighting scheme for Holm's sequentially rejective alpha-adjustment procedure. Investigation of the correlation between endpoints is examined in relation to adjustment of the alpha-level. In the context of a clinical trial, the above multiplicity procedures are applied and compared. Finally, some comments concerning ease of use and relevance are summarized for the above methods. PMID- 9204222 TI - Interpreting tests for efficacy in clinical trials with multiple endpoints. PMID- 9204223 TI - Outcomes of a placebo run-in period in a head and neck cancer chemoprevention trial. AB - This study describes the outcomes of an eight-week placebo run-in period in a head and neck cancer chemoprevention trial. Of 391 former cancer patients who entered the run-in over the first two years of the trial, 91% were randomized. Pill counts showed that adherence rates ranged from 0% to 120% (mean 96%, SD = 15%). The trial did not randomize subjects who were no longer interested in trial participation (n = 20), who did not return within 10 weeks of enrollment date (n = 3), or who did not achieve a drug adherence level of at least 75% (n = 9). Three subjects were not randomized for other reasons. Univariate predictors of run-in outcome (randomized or not randomized) included ethnicity, education level, cancer site, cancer stage, and Karnofsky performance score. Multivariate analyses resulted in a logistic model with Karnofsky performance and education level as significant predictors of randomization. Persons with a Karnofsky score of 100 had 2.3 higher odds of randomization (95% CI = 1.1, 4.9) than persons with compromised Karnofsky scores, and persons with more than a high school education had 2.1 higher odds of randomization (95% CI = 1.0, 4.9) than persons with less education. These results suggest that the use of a run-in period may compromise the external validity of randomized prevention trials. More research is needed to understand further the behavioral factors underlying the observed differences so that prevention researchers can develop effective interventions for facilitating trial participation, especially in under-represented, trial-eligible groups. Investigators should expand the objectives of a run-in period to (1) evaluate why eligible persons refuse trial enrollment or fail to be randomized at the end of the run-in and (2) use the run-in period for a systematic evaluation of levels and costs of intervention strategies designed to promote trial enrollment and adherence. PMID- 9204224 TI - Assessment of a food frequency questionnaire as a screening tool for low fat intakes. AB - This study tested the ability of a self-administered food frequency questionnaire (FFQ) to identify women with low fat intakes. FFQs were completed by 95 control participants of a dietary trial at a mean 2.9 +/- 0.8 years post-randomization. Subjects were selected in approximately equal numbers from women who were low-fat eaters (< or = 30% of energy from fat) and high-fat eaters (> 30% of energy from fat). Percentage energy from fat derived from food records and FFQ were similar in both the low- and high-fat eaters. Percentage of energy from carbohydrate and total grams of carbohydrate (low-fat eaters only) were slightly higher measured by FFQ than by food records, and percentage of energy from protein was slightly lower. The correlation between nutrient intake measured by FFQ and food records for the whole group was 0.74 for percentage of energy from fat, 0.50 for total fat, 0.59 for percentage of energy from carbohydrate, 0.43 for total carbohydrate, 0.53 for percentage of energy from protein, 0.27 for total protein, and 0.32 for energy intake. Correlations were slightly lower when the low- and high-fat eaters were examined separately. The area under the receiver operating characteristic (ROC) curve, 0.83, was significantly above 0.5 (p < < 0.001), indicating that the FFQ discriminated between low- and high-fat eaters significantly better than chance. The FFQ cutoff point of 30% of energy from fat had a true positive rate of 0.63 and false positive rate of 0.24. The use of this cutoff point for screening would result in the loss of 36% of potential subjects and an estimated increase in baseline percentage of energy from fat intake of 2.3 percentage points. PMID- 9204226 TI - Fetal testing in postdates. AB - Postdate pregnancies are at significant risk for perinatal morbidity. Fetal testing may be used to observe the prolonged pregnancy safely while awaiting onset of labor. Pitfalls associated with the use of fetal testing in this situation are discussed. The current literature addressing these problems is reviewed. PMID- 9204225 TI - Use of a stochastic simulation model to identify an efficient protocol for ovarian cancer screening. AB - The intervention protocol for an ovarian cancer screening trial should be efficient as well as effective, because it may become the standard of care if the trial demonstrates mortality reduction. To identify an efficient ovarian cancer screening protocol, the effectiveness and cost-effectiveness of selected single modality and multimodal screening strategies were estimated using a stochastic simulation model. Screening was simulated over a 30-year period in a hypothetical cohort of 1 million women aged 50 at the beginning of the period. The net present value of the cost per year of life saved was estimated for six protocols involving transvaginal sonography (TVS) and/or the tumor antigen CA 125. Internal and external validation was performed, and sensitivity analyses were conducted to assess the robustness of the ranking of the strategies. A multimodal strategy involving CA 125 with a threshold for positivity of either elevation above 35 U/ml or doubling since the previous screen, followed by TVS only if CA 125 is positive, was found to be efficient in the sense that no other strategies saved as many years of life at lower cost per year of life saved. Used annually, this strategy cost under $100,000 per year of life saved over a range of assumptions. The model's predictions are consistent with results reported in the literature regarding the performance of TVS and CA 125. The multimodal strategy used annually or every six months was efficient compared to either ultrasound or CA 125 used alone, over a range of assumptions. Simulation of screening may be useful in selecting a screening protocol to be tested in a randomized controlled trial. PMID- 9204227 TI - Fetal surveillance in diabetic pregnancy. AB - The focus of monitoring in diabetic pregnancy is no longer the prevention of fetal mortality, owing to the impressive benefits of strict maternal glucose control. Against this background, fetal monitoring must account for congenital anomalies, fetal mortality and severe morbidity as a result of metabolic consequences of hyperinsulinism, the exponential effect of diabetes when other maternal complications are present, and peripartum problems of the macrosomic infant, of delayed lung maturation, birth trauma and neonatal hypoglycemia. Thus, a broad range of potential fetal problems with varying maternal complications requires individualized, serial observation with multiple-format, properly validated tests. There has been recent progress: definition of the population at risk, clarification of pathophysiology, application of multiple-format tests, and evaluation of neonatal impacts; further precision may be developed with specific fetal tests. This review deals with the continued fine tuning of this critical area of perinatal medicine. PMID- 9204229 TI - Fetal testing in twins. AB - The incidence of twin pregnancies continues to rise with increasing use of assisted fertility techniques. The search for accurate methods of chorionicity determination and aneuploidy screening, and the outcome following multi-fetal pregnancy reduction dominate the recent literature. In addition, debate on the aetiology and management of the transfusional complications of monochorionicity continues. PMID- 9204228 TI - Fetal testing in intra-uterine growth retardation. AB - There are three major issues that remain subject to debate in relation to fetal growth retardation: (1) Which method should be used to identify the affected population? (2) Which biophysical test(s) is most appropriate to assess the pregnancy? (3) Which factors are important when considering elective delivery? This review summarizes recent studies investigating the efficacy and accuracy of tests used for the identification and surveillance of pregnancies at risk of growth retardation, and discusses the importance of gestational age in the decision for or against intervention. PMID- 9204230 TI - Epidemiology of antepartum fetal testing. AB - Advances in perinatal and neonatal health care over the past few decades have resulted in a substantial reduction in perinatal mortality. Some of this improvement has been attributed to antepartum fetal surveillance techniques. The primary objective of antepartum fetal surveillance techniques is to avoid fetal deaths. An ideal secondary objective is to avoid neonatal complications related to intrauterine asphyxia. In this article, some of the difficulties in evaluating existing antepartum fetal surveillance techniques are highlighted. Some of the epidemiological methods for evaluating a screening test are reviewed and their importance discussed with reference to fetal testing procedures. Lastly, the possibility of considering indication-specific fetal testing to improve perinatal morbidity is examined. PMID- 9204231 TI - Prenatal diagnosis. PMID- 9204232 TI - Antenatal genetic counselling: implications for population screening. AB - Only 1-2% of newborns have a congenital abnormality, yet it is responsible for the much greater proportion of mortality and morbidity in infancy and in childhood as well as during pregnancy. Because of this prevalence, there have been many developments in screening and diagnosis of congenital abnormality. Yet there are a number of limitations about how the screening and diagnostic service is currently operated. These limitations centre on the information that is provided, who undertakes the counselling, and their training and support provided to parents in making important decisions. PMID- 9204233 TI - Neural tube defects/alpha-fetoprotein/Down's syndrome screening. AB - Maternal serum screening for alpha-fetoprotein, initially as an indicator of open neural tube defects and more recently combined with other pregnancy-specific markers as an indicator of Down's syndrome, is an established part of prenatal testing. Recent work has focused on the role of folic acid in the primary prevention of neural tube defects and on the search for a genetic component in the aetiology of neural tube defects. Potential new markers of Down's syndrome have been identified in maternal serum and urine. Nuchal translucency measurements by ultrasound and the identification of appropriate maternal serum markers in the first trimester offer the prospect of screening earlier in pregnancy. PMID- 9204234 TI - Progress in the genetic analysis of fetal cells circulating in maternal blood. AB - The presence of fetal cells in maternal blood has been demonstrated by many investigators who continue to refine methods to permit their isolation, identification and genetic analysis. The sensitivity and specificity of this technique for noninvasive aneuploidy detection is currently being addressed in a multicenter clinical trial. Significant advances have also occurred in fetal single-cell and single-gene analysis. PMID- 9204235 TI - Mosaicism: implications for postnatal outcome. AB - Mosaicism detected in the cytogenetic analysis of chorionic villi or cultured amniocytes can present a difficult and at times impossible interpretative dilemma. The finding may be indicative of a generalized fetal mosaicism. However, in the majority of cases, the abnormal cell line is representative of either an in-vitro event or a chromosomal error that is restricted to the extra-embryonic tissues. Advances in laboratory medicine have provided insight into the determination of which cases of true mosaicism have the greatest risk of being clinically significant with regard to the fetal genotype and phenotype. Despite the presence of a normal fetal karyotype, certain chromosomes involved in confined placental mosaiciam may increase the risk of poor perinatal outcome or predispose the fetus or neonate to the adverse effects of genetic imprinting owing to the development of uniparental disomy. PMID- 9204236 TI - Congenital abnormalities in twins: selective termination. AB - Selective termination for an anomalous twin in discordant twin pregnancy has been a management option for nearly two decades. Using current conventional prenatal diagnostic techniques, the diagnosis of a twin gestation discordant for a congenital anomaly is commonly confirmed in the second trimester. Selective termination is thus usually a second-trimester procedure. The risks of second trimester terminations as well as protocol choices, including indications and timing, have evolved over the years. Ethical issues regarding selective termination have developed significantly and continue to be debated. PMID- 9204237 TI - Severe ovarian hyperstimulation syndrome: serum and ascitic fluid concentrations of vascular endothelial growth factor. PMID- 9204238 TI - Maternal-fetal medicine and prenatal diagnosis. PMID- 9204239 TI - Prediction of outcome after perinatal asphyxia. AB - Perinatal asphyxia at term remains a significant cause of infant death and neurodevelopmental impairment, probably causing 20% of all cases of cerebral palsy. This review examines indicators that can help determine the prognosis after suspected asphyxia in term infants, including obstetric information, clinical examination, and diagnostic methods based on neurophysiology, neuroimaging, and biochemistry. New data confirm that many frequently used indicators are generally not specific predictors, while simple electroencephalographic methods, magnetic resonance spectroscopy, and magnetic resonance imaging can have high positive predictive values even when used quite soon after birth. PMID- 9204240 TI - Neonatal infections. AB - Neonatal infections range from early-onset vertically acquired infections to nosocomial bacterial sepsis, and they account for significant morbidity and mortality globally. The recent emergence of perinatally acquired HIV-1 infection has also placed an enormous burden on existing meager health resources in developing countries. The risk of early-onset group B streptococci sepsis may be related to heavy genital tract colonization at term. Perinatally acquired HIV-1 infection also occurs at birth, with higher rates of infection after vaginal delivery. Detection of circulation HIV-1 viral DNA by polymerase chain reaction offers an extremely sensitive screening method in the neonatal period. In contrast, rapid confirmation of bacterial sepsis in the newborn still poses considerable problems. A review of data on antimicrobial prophylaxis of preterm premature rupture of membranes does reveal improved neonatal outcome. However, the rapid and widespread emergence of multidrug resistance among bacterial pathogens dictates the need for judicious use of antibiotics, as well as close attention to preventive strategies. PMID- 9204241 TI - The present and future role of gene therapy in the newborn. AB - There are currently a large number of ongoing gene therapy trials in North America and Europe. These trials have almost exclusively involved patients with inherited lethal disorders or malignancies. Although there are significant ethical and safety issues that remain unresolved, it seems inevitable that this technology will soon be adapted for use in lethal, or potentially lethal, fetal and neonatal diseases. If ethical and safety issues can be resolved, a wide spectrum of nonlethal acute and chronic diseases could also benefit from this form of therapy. The purpose of this brief review is to provide an overview of current approaches to gene delivery, their successes, and their limitations. Where possible, the discussion has focused on conditions that are recognized in fetal or neonatal life, to give the reader some sense of the potential scope for this form of therapy. PMID- 9204242 TI - Fetal and neonatal nephrology. AB - In this review, the recent literature pertaining to the nephrology of the fetus and neonate is reviewed. Some papers of interest are described, and an overview of active areas of research is given. One of these areas is early development, studies of which are giving new insight into patterns of normal and abnormal kidney morphogenesis. Preterm newborn infants may suffer from electrolyte imbalance and poor growth as a result of tubular dysfunction, and this subject is discussed. Also addressed are some advances in genetics that are elucidating the genetic defects of cystic kidney diseases and providing clues about the pathogenesis of renal dysplasia. Other areas discussed in this review are the assessment of renal function in the fetus and neonate, blood pressure and hypertension in the neonate, renal failure in the term and preterm infant, and the causes and the consequences of fetal urinary obstruction. PMID- 9204243 TI - Glomerulonephritis in childhood. AB - Safer renal biopsy techniques have led to increased recognition of the various forms of glomerulonephritis in the pediatric population. Our understanding of their natural history and progression has improved, and we now know that there is significant morbidity associated with diseases such as IgA nephropathy and membranoproliferative glomerulonephritis. Knowledge of the pathophysiology of progressive renal disease has also expanded, but specific treatment modalities, especially for children, are lacking and continue to be areas for future clinical research. This article reviews four types of glomerulonephritis that occur in childhood: acute poststreptococcal glomerulonephritis, IgA nephropathy, Alport's syndrome, and membranoproliferative glomerulonephritis. The clinical and pathologic features of each are reviewed, and the current literature covering new developments in their prognosis, genetics, or therapies are summarized. PMID- 9204244 TI - Nephrocalcinosis. AB - The routine use of ultrasonography has resulted in an explosion in the number of conditions reported to be associated with nephrocalcinosis. It has also been increasingly recognized that urolithiasis and nephrocalcinosis can coexist in the same patient. The two conditions most commonly associated with nephrocalcinosis in childhood are the use of furosemide in infancy and the treatment of patients with hypophosphatemic rickets with phosphate and vitamin D preparations. Although originally thought to be related to hypercalciuria, more recent studies in humans and research animals indicate a multifactorial etiology for furosemide-related nephrocalcinosis. In patients with hypophosphatemic rickets, it seems that the dose of phosphate and in particular the development of secondary hyperparathyroidism play a central role in the development of nephrocalcinosis. Among the entities recently reported to be associated with nephrocalcinosis are some that characteristically include Fanconi's syndrome. These findings dispute the previous teaching of lack of renal calcifications in this syndrome, which involves proximal renal tubular acidosis. The diagnosis of nephrocalcinosis requires a metabolic work-up to identify the offending factor. When identified, appropriate intervention should be instituted. PMID- 9204245 TI - Toxic nephropathies. AB - A variety of medications can adversely affect the kidney. Awareness of these potential nephrotoxic effects is crucial to prevention of serious sequelae. This review discusses renal injury caused by the chemotherapeutic agent ifosfamide, the problem of analgesic nephropathy, and the newly identified complication of trimethoprim-induced hyperkalemia. In addition, recent information about the classic nephrotoxins (cisplatin, amphotericin B, and aminoglycosides) is presented. PMID- 9204246 TI - Lead poisoning in children. AB - Increased lead exposure and increased body burden of lead remains a significant problem for children in the United States. With the increased use of blood level screening methods, a large percentage of children in many industrialized countries are being tested as a being at risk. A controversy continues over the definition of what population to screen and at what age to screen. There are parts of the United States, especially rural areas and health maintenance organization populations, where screening for lead exposure has not been productive. A new drug, DMSA (meso 2,3-dimercaptosuccinic acid) has been approved for oral chelation of children with increased body burden of lead. At the present time it is labeled for use in children with blood lead concentrations in excess of 45 micrograms/dL. Evidence exists that DMSA is effective in lowering the blood lead concentrations in children with levels between 25 and 45 micrograms/dL. The long-term effectiveness of chelation at lower levels is at present uncertain. There remains no substitution for strict environmental decontamination in the home environment of children and the workplace environment of their parents. PMID- 9204247 TI - Antiviral chemotherapy. AB - Over the last few years, several new antiviral agents have been added to the chemotherapeutic armamentarium. This review discusses advances in the treatment of HIV disease in children and adults. More pronounced antiviral effects have been achieved through the study of new agents such as the protease inhibitors as well as through a better understanding of previously approved drugs, such as reverse transcriptase inhibitors. PMID- 9204248 TI - Postnatal steroid therapy in neonates. AB - Dexamethasone treatment of neonates to facilitate weaning from mechanical ventilation has become increasingly common. Despite thousands of infants being treated, most studies fail to demonstrate an improvement in long-term pulmonary and neurodevelopmental outcome. Much controversy remains regarding who should be treated, when treatment should begin, what the proper dose is, how long treatment should continue, and how adverse effects can be minimized. Early treatment prior to the onset of significant pulmonary inflammation may be an approach that improves outcomes and allows for shorter treatment courses. Risk assessment tools using ventilator requirements, adrenal function, or markers of pulmonary inflammation may allow for improved patient selection. Future studies will need to include a large number of subjects and will need to be continued beyond the neonatal period to assess long-term pulmonary and neurodevelopmental outcome. PMID- 9204249 TI - Autism, child abuse, and sudden infant death syndrome. AB - The current literature regarding the standard and nonstandard therapies for children with autism is reviewed. A long term, comprehensive, individualized, multidisciplinary approach remains the best treatment. Physicians caring for the victims of child abuse are frequently asked to render an opinion regarding soft tissue bruising. A review of the literature suggests that estimation of the age of a bruise should not rely solely on color, but rather should be the result of careful history, a through physical examination, and possibly laboratory testing. The need for a standardized and systematic approach to sudden infant death syndrome is also reviewed. The psychological effects on the parents following sudden infant death is discussed and reveals maternal anxiety and depression and, to a lesser degree, paternal anxiety and depression following the loss of a child. Currently, sleep position continues to be a risk factor for sudden infant death syndrome, although immunizations may not be. PMID- 9204250 TI - Clinical therapeutics. PMID- 9204251 TI - Biologic effects of nonsteroidal anti-inflammatory drugs. AB - The nonsteroidal antiinflammatory drugs (NSAIDs) continue to be important therapeutic interventions for the treatment of pain and inflammation. Investigators continue to produce new information about their biologic effects, including their actions as well as their toxic effects and methods to decrease the potential side effects associated with their use. This year many papers have been published speculating about those NSAIDs that are reported to selectively inhibit the isoform of the cyclooxygenase enzyme that is induced in inflammatory conditions (Cox-2) rather than that associated with normal physiologic function (Cox-1). Reports have shown that the more Cox-2-selective NSAIDs (from three- to 10-fold more selective for Cox-2 over Cox-1) seem to have less gastrointestinal toxicity associated with their use; however, little evidence has yet emerged from phase I, II, or III studies about the clinical effects of the highly selective Cox-2 inhibitors (300-fold or more selective for Cox-2 over Cox-1). In addition, intriguing animal studies have shown the effects of knockout of the genes controlling the activities of either Cox-1 or Cox-2 in mice. PMID- 9204252 TI - Disease-modifying antirheumatic drugs. AB - The case for early intervention with disease-modifying antirheumatic drugs is strengthened by published reports during the past year. These drugs include methotrexate, gold sulfasalazine, and antimalarial agents. The American College of Rheumatology issued guidelines for the management of rheumatoid arthritis and for monitoring the toxicity of antirheumatic drugs. Studies on the mechanisms of action of disease-modifying antirheumatic drugs focused on their effects on cytokines and their receptors. The toxic effects of disease-modifying antirheumatic drugs remained an important issue, especially the adverse pulmonary effects of methotrexate. An important trial demonstrated a beneficial effect of triple disease-modifying antirheumatic drug therapy (methotrexate, sulfasalazine, and hydroxychloroquine) over individual agents. PMID- 9204253 TI - Immunosuppressive drug treatment. AB - Use of the immunosuppressive agents in patients with rheumatic diseases is becoming increasingly common as our experience using these agents increases. This year's review of recent reports on immunosuppressive drugs for rheumatic diseases focuses on recent contributions to the literature regarding individual agents. Additionally, we review recommendations regarding immunization of immunosuppressed patients and exposure of such patients to common communicable illnesses to which they may be inadvertently exposed. PMID- 9204254 TI - Corticosteroids in the treatment of rheumatologic diseases. AB - Despite continued reports of their efficacy in certain circumstances, controversy over the role of corticosteroids in the treatment of the rheumatologic diseases, especially of rheumatoid arthritis, persists. New clinical data are emerging on the role of corticosteroids in combination with other antirheumatic drugs and about the local use of corticosteroids in other inflammatory arthropathies. In the laboratory, understanding of the effect of corticosteroids on the phenotype of synovial leukocytes has been enhanced. The effects of corticosteroids on bone mineral density continue to be studied, although large, well-controlled, comparative studies are still few in number, and use of osteoprotective regimens in the community has been shown to be low. PMID- 9204255 TI - Biologic agents in the treatment of inflammatory rheumatic diseases. AB - In 1996, experience in treating autoimmune rheumatic diseases with biologic agents has further improved. New data have been published using different principles directed against cell surface antigens or against proinflammatory cytokines or applying anti-inflammatory cytokines such as interleukin-10 and interleukin-4. In addition, combination therapies with anti-tumor necrosis factor alpha monoclonal antibody and methotrexate have shown sustained long-lasting beneficial effects. Undoubtedly, with increasing knowledge of the exact mechanisms leading to tissue destruction in autoimmune rheumatic diseases, new treatment principles will be developed that may be even more specific and may be associated with fewer adverse effects than the biologic agents tested thus far. PMID- 9204256 TI - Cellular pathways of joint destruction. AB - In the past year, evaluation of cellular pathways of rheumatoid synovial fibroblasts has been the focus of several laboratories investigating the molecular and cellular mechanisms operating in the pathogenesis of rheumatoid arthritis. Major topics that have been reported on include the adhesion and interaction of synovial fibroblasts with cartilaginous matrix and other synovial cells, intracellular signaling, and activation of gene transcription. Novel approaches to inhibiting cartilage destruction have also been described. In the latter case, thorough multicenter characterization of the interleukin 1/interleukin-1 receptor antagonist pathways resulted in the first gene therapy trials in animals and humans. PMID- 9204257 TI - Lessons for joint destruction from animal models. AB - There is growing evidence that tumor necrosis factor-alpha is involved in onset of arthritis, whereas the cartilage destructive process is mainly interleukin-1 driven. Moreover, direct generation of interleukin-1 may occur in the absence of tumor necrosis factor action. This is shown in various arthritis models using neutralizing antibodies, receptor antagonists, and soluble receptors, and similar proof has been provided in cytokine-targeted transgenic and knockout mice. Further evidence for local impact of interleukin-1 is obtained from gene transfer of interleukin-1 receptor agonist to joint tissues, whereas these experiments also underline the therapeutic applicability of this approach. In addition to tumor necrosis factor-alpha and interleukin-1 action, the destructive process appears to be under the control of mediators such as interleukin-6 and -10. Marked cartilage protection can be achieved with additional treatment with interleukin-4 and -10. These modulatory studies also underline the potential uncoupling of inflammatory and destructive processes. Although T cells may drive arthritic processes, it is clear that fibroblasts can cause major destruction in the absence of T cells. Insight in pivotal enzymes primarily involved in cartilage destruction is still limited. PMID- 9204258 TI - Management of rheumatoid arthritis. AB - There is increasing evidence that reduction of disease activity by disease modifying drugs alters the disease course of rheumatoid arthritis and that patients benefit from early introduction of disease-modifying antirheumatic drugs. To ensure accurate diagnosis and timely referral, training of medical students and residents in rheumatology and access to a physician experienced in the management of rheumatic diseases is important. Because spontaneous remission is rare, the risk of overtreatment of a few patients is outweighed by the benefits of early treatment for the majority of patients. Also, side effects may be reduced with optimation of titration of disease activity based on standardized, reliable, valid, sensitive, and easily interpretable measures with clear-cut decision rules. A potential solution to the practical problems involving the scoring of patient questionnaires such as the Health Assessment Questionnaire and the Rheumatoid Arthritis Disease Activity index and clinical algorithms such as the Disease Activity Score is the provision of a feedback system similar to a laboratory. To feed back the results of titration of disease activity with the goal of permanently improving health outcomes in an ongoing learning process may be called clinical quality management. From a health care system or political perspective, clinical quality management that demonstrates the commitment of rheumatology toward continuous improvement of rheumatoid arthritis care may become an important activity of rheumatology societies in countries where specialty care is under scrutiny and where specialists are increasingly required to show the benefits of their care. PMID- 9204259 TI - T-cell-dependent pathways in rheumatoid arthritis. AB - Two theories, not necessarily mutually exclusive, attempt to explain the pathogenesis of rheumatoid arthritis. The mesenchymal hypothesis proposes that, after an initial event triggered by T cells, synovitis is maintained by autocrine and paracrine pathways involving macrophages and synovial fibroblasts. The T-cell hypothesis proposes that T cells are continuously involved in the pathogenesis of rheumatoid arthritis from its initiation phase through to the chronic stage. This paper reviews recent work in this area and concludes that there is substantial evidence in support of the T-cell hypothesis. PMID- 9204260 TI - Mechanisms of cartilage destruction and novel nonsurgical therapeutic strategies to retard cartilage injury in rheumatoid arthritis. AB - Although there are excellent rationales for the use of biologic agents, no published novel therapeutic strategy in patients with early or late rheumatoid arthritis has thus far been proven in controlled clinical studies to prevent or retard cartilage destruction. Although T-cell-specific therapies in chronic rheumatoid arthritis have some success, the percentage of patients responding and the degree of clinical improvement are disappointing, and cartilage injury can neither be prevented nor retarded. Similarly, attempts to interrupt the cytokine loops and inhibit adhesion molecules are only modestly successful. The alternative approach of solely controlling the effector side by direct or indirect metalloproteinase inhibition seems attractive because it would circumvent the cytokine networks while theoretically still preventing the final consequences of inflammation on cartilage. One therapeutic strategy that inhibits metalloproteinses-tetracyclines or chemically transformed tetracyclines-cannot be considered a breakthrough with respect to either reduction of disease activity or prevention or retardation of cartilage injury in rheumatoid arthritis. It is likely that combination therapy will be further developed in the future. The most promising agents today, such as the anti-tumor necrosis factor-alpha antibodies, must be combined with other current strategies as well as with newly developed disease-specific biologic agents. To affect multiple sites in the underlying inflammatory process and to target the delivery of the agents could be one of the goals. The efficacy and effectiveness of any new strategy depends on its ability to alter the function of the aggressive and transformed synovial fibroblasts within the pannus and to protect the articular chondrocytes and early cartilage injury. PMID- 9204261 TI - Crystal deposition disease. PMID- 9204262 TI - Impaired proprioception and osteoarthritis. AB - The increase in the prevalence of osteoarthritis (OA) with age may be due in part to increased joint load resulting from age-related declines in neuromechanical factors, including joint position sense or proprioception. Several studies have demonstrated that knee proprioception is worse in knee OA patients versus age matched control subjects. Functional consequences of impaired proprioception may include lower gait velocity, shorter stride length, and slower stair walking time. Some studies have shown that proprioception can be enhanced by wearing an elastic bandage or similar orthoses and by muscle training. A variety of other interventions have been proposed as well. To date studies of proprioception in OA patients have been cross-sectional. Theoretically, impaired proprioception might contribute toward or result from OA. A paradigm depicting directions in the relationship between proprioception impairment and OA is offered. Longitudinal data are badly needed to elucidate cause and effect and to determine the relative importance of impaired proprioception in disease progression. The relationship between sensory input and protective or damaging muscle activity has been minimally evaluated in the setting of OA. In studies of clinical conditions related to OA, experimental effusions had no effect on proprioception. Proprioception was worse in hypermobility syndrome patients versus age-matched controls. Anterior cruciate ligament insufficiency is associated with a decline in proprioception. PMID- 9204263 TI - Muscle weakness in osteoarthritis. AB - Muscle has an integral role in the structure and function of joints. Evidence for muscle weakness in osteoarthritis of the knee exists and is not fully explained by the effects of aging. Weakness is associated with pain and disability. The temporal relationship requires further study, but preliminary evidence for a causative role is emerging. Muscle weakness can be assessed in various ways. Voluntary measures of strength are affected by degree of effort. In osteoarthritic patients, as in other patient groups, effort may be influenced by pain and psychologic outlook. Techniques to estimate muscle activation are available but have yet to be fully explored in osteoarthritis. Quadriceps exercises increases strength and may have beneficial effects on pain and function. The long-term benefits of exercise for therapy and possible prevention of osteoarthritis are not yet known. This is an exciting area of research, and it is anticipated that more findings will emerge in the near future. PMID- 9204264 TI - Plain radiography in the evaluation of knee osteoarthritis. AB - The prospects for results of controlled studies of disease-modifying osteoarthritis drugs in humans have fostered interest in radiographic methods to detect early cartilage damage and to assess progressive cartilage changes in osteoarthritis of the knee. We summarize here the extent to which technical factors in plain knee radiography-ie, the degree of knee flexion, misalignment of the x-ray beam, magnification of the radiographic image of the joint-and mensural procedures can impact the reproducibility of quantitative measurements of tibiofemoral joint space width, the surrogate for thickness of articular cartilage. Failure to standardize crucial elements of conventional knee radiography and to computerize measurement of tibiofemoral joint space width will introduce significant (and probably insurmountable) error variation into radiographic outcome data. The level of precision achievable with fluoroscopically-assisted flexion of the knee and rotation of the foot with computerized, magnification-corrected measurement of joint space width greatly improves the feasibility of disease-modifying osteoarthritis drug trials as they relate to sample size and duration of treatment necessary to detect an effect from disease-modifying osteoarthritis drugs. PMID- 9204266 TI - Calcium crystal-associated diseases and miscellaneous crystals. AB - Calcium crystal-associated diseases are still a challenge in clinical and basic science. Advances in understanding crystal formation and dissolution and crystal participation in inflammation are reported here. Studies on different methods of identification are reviewed, emphasizing the need for accurate and reproducible ways to study and diagnose calcium crystal-associated diseases. Reports of uncommon presentations are also described, including a controlled study on calcification of the ligamenta flava of the spine and a study involving 19 years of clinical follow-up of families with hydroxyapatite chondrocalcinosis with spondyloepiphyseal dysplasia. Studies on miscellaneous crystals are few. Two recent reviews are described on cholesterol crystal embolization syndrome and cryocrystalglobulinemia. PMID- 9204265 TI - Gout and hyperuricemia. AB - Although there continue to be relatively few basic studies of the metabolism and transport of urate, clinical interest in gout and hyperuricemia remains high. A number of interesting new observations are described here from the published literature of the past year. Various aspects of the diagnosis, management, and pathophysiology of gout and gouty arthritis have been recently reviewed. The therapy of gout and hyperuricemia continue to lead to significant complications. PMID- 9204268 TI - Rheumatoid arthritis. PMID- 9204267 TI - Clinical therapeutics. PMID- 9204269 TI - Osteoarthritis and crystal deposition diseases. PMID- 9204271 TI - RNA structure: crystal clear? AB - Structured RNAs play an essential role in chromosome maintenance, RNA processing, protein biosynthesis, and protein transport. To understand RNA function in these diverse biological systems, the rules for RNA folding and recognition must be learned. Recent crystal structures of hammerhead ribozymes, a group I intron domain, and RNA duplexes provide new insights into the principles of RNA folding and function. PMID- 9204270 TI - Nucleic acids. From self-assembly to induced-fit recognition. PMID- 9204272 TI - Recent solution structures of RNA and its complexes with drugs, peptides and proteins. AB - The past two years have seen remarkable progress in the study of RNA structure: the predicted era of RNA structural biology has arrived. Crystallographic structures of the hammerhead ribozyme and of a large subunit of a group I self splicing intron have begun to reveal the structural basis of RNA enzymatic activity. A remarkable number of structures of small RNAs and of complexes with drugs, peptides and one protein domain have been determined by NMR. PMID- 9204274 TI - Protein-facilitated RNA folding. AB - In the absence of protein collaborators, both simple and complex RNAs often misfold or are unfolded. Biologically important RNAs solve their folding problem, in part, using the assistance of chaperone and cofactor proteins. Recent work emphasizes several rules for RNA-protein complexes: formation involves induced fit; many large RNAs fold slowly; and ribonucleoprotein assembly requires multiple steps. Finally, protein binding can introduce thermodynamic side effects. PMID- 9204273 TI - The New World of ribozymes. AB - The number of RNA molecules that have novel catalytic activities has dramatically increased during the past two years. This ribozymic boom is not due to the discovery of additional examples of natural ribozymes but rather to the development of artificial ribozymes isolated by in vitro selection and evolution techniques. The structural and functional complexities of these artificial ribozymes, however, do not match those of the larger natural ribozymes. The understanding of both RNA structure and catalysis performed by natural and artificial ribozymes paves the way for the creation of RNA molecules that are able to efficiently catalyze more complex reactions. PMID- 9204275 TI - The conformation of ribosomes and rRNA. AB - During the past 18 months, electron microscopists have published two reconstructions of the Escherichia coli ribosome, independently derived from images of unstained particles. The resolutions of their images are 20-25 A-much higher than any previously available. During the same time, NMR spectroscopists have provided an atomic-resolution model for the A-site region of 16S rRNA complexed with paromomycin that explains much of what is known about the interaction of aminoglycoside antibiotics with ribosomes. PMID- 9204276 TI - Unusual DNA conformations. AB - DNA is on the move across conformational space. Duplexes diversity and, joined by triplexes, quadruplexes, loops, bulges and multiarmed junctions, open the route to a bewildering array of increasingly complex conformations. In addition to this structural growth, DNA has come under increasing scrutiny thanks to the development of chemical and physical techniques for deforming its conformation and probing its properties. These investigations help us to learn more about the mechanics and the activity of this remarkably versatile macromolecule. PMID- 9204277 TI - Targeting the minor groove of DNA. AB - Small molecules that specifically bind with high affinity to any predetermined DNA sequence in the human genome will be useful tools in molecular biology and, potentially, in human medicine. Pairing rules have been developed to control rationally the sequence specificity of minor groove binding polyamides containing N-methylimidazole and N-methylpyrrole amino acids. Using simple molecular shapes and a two-letter aromatic amino acid code, pyrrole-imidazole polyamides achieve affinities and specificities comparable to DNA-binding proteins. PMID- 9204278 TI - Visualizing nucleic acids and their complexes using electron microscopy. AB - Microscopic visualization of nucleic acid-protein complexes provides a means of obtaining structural information that is difficult to obtain in any other way, and of verifying conclusions derived from other approaches. The polymorphic, flexible, and irregular nature of these complexes presents particular problems in their analysis. PMID- 9204279 TI - Population statistics of protein structures: lessons from structural classifications. AB - Structural classifications aid the interpretation of proteins by describing degrees of structural and evolutionary relatedness. They have also recently revealed strikingly skewed distributions at all levels; for example, a small number of folds are far more common than others, and just a few superfamilies are known to have diverged widely. The classifications also provide an indication of the total number of superfamilies in nature. PMID- 9204280 TI - Progress in protein structure prediction. AB - If protein structure prediction methods are to make any impact on the impending onerous task of analyzing the large numbers of unknown protein sequences generated by the ongoing genome-sequencing projects, it is vital that they make the difficult transition from computational 'gedankenexperiments' to practical software tools. This has already happened in the field of comparative modelling and is currently happening in the threading field. Unfortunately, there is little evidence of this transition happening in the field of ab initio tertiary structure prediction. PMID- 9204281 TI - Predicting coiled-coil regions in proteins. AB - The past several years have seen significant advances in our ability to recognize coiled coils from protein sequences and model their structures. New methods include a detection program based on pairwise residue correlations, a program that distinguishes two-stranded from three-stranded coiled coils and a routine for modelling the coordinates of the core residues in coiled coils. Several widely noted predictions, among them those for heterotrimeric G proteins and for cartilage oligomeric matrix protein, have been confirmed by crystal structures, and several new predictions have been made, including a model for the still hypothetical right-handed coiled coil. PMID- 9204282 TI - Prediction of N-terminal protein sorting signals. AB - Recently, neural networks have been applied to a widening range of problems in molecular biology. An area particularly suited to neural-network methods is the identification of protein sorting signals and the prediction of their cleavage sites, as these functional units are encoded by local, linear sequences of amino acids rather than global 3D structures. PMID- 9204283 TI - Searching for regulatory elements in human noncoding sequences. AB - Important progress has been made in the past two years in the identification of Pol II promoters. For most other regulatory elements, however, current biological knowledge is still insufficient to allow the development of prediction tools. The phylogenetic-footprinting strategy, which is based on the comparative analysis of homologous sequences, is a very efficient approach to identify new unknown regulatory elements. The recent organization of large-scale sequencing projects for some model vertebrate organisms will be extremely valuable for the prediction of regulatory elements in the human genome. PMID- 9204284 TI - Modular multidomain phosphoryl transfer proteins of bacteria. AB - Recent phylogenetic and structural analyses of multidomain phosphoryl transfer proteins of bacteria have revealed that interdomain (but not intradomain) splicing and fusion, as well as domain duplication and deletion, have occurred frequently during evolution. These events have been found to be exceedingly rare in certain other protein families. Domain-shuffling events are illustrated by examples from the superfamilies of phosphoenolpyruvate-dependent sugar phosphotransferase systems, their transcriptional regulatory protein targets of phosphorylation, sensor autokinase/response regulator signal transduction systems, and permeases of the ATP-binding-cassette type. PMID- 9204285 TI - Three-dimensional domain duplication, swapping and stealing. AB - Examination of multidomain and/or multimeric protein structures can reveal evolutionary paths to a more complex 3D organization. Over the past few years, proteins have been shown to evolve while preserving mutual domain organization and interfaces. The recent advances in understanding domain reorganization and mobility highlight the versatility and efficiency of protein structural evolution. PMID- 9204286 TI - Circular permutations of natural protein sequences: structural evidence. AB - Over the past few years, evidence has accumulated that shows that circularly permuted proteins resulting from permutations in their coding genes can indeed occur naturally. In most instances, these circularly permuted amino acid sequences have been detected by sequence alignment of homologous proteins. Circular permutations may escape detection, however, when based on sequence comparisons alone, as recently illustrated by transaldolase, a member of the class I aldolase family. PMID- 9204287 TI - Nucleic acids. PMID- 9204288 TI - Sequences and topology. PMID- 9204289 TI - The economic impact of a bioterrorist attack: are prevention and postattack intervention programs justifiable? AB - Understanding and quantifying the impact of a bioterrorist attack are essential in developing public health preparedness for such an attack. We constructed a model that compares the impact of three classic agents of biologic warfare (Bacillus anthracis, Brucella melitensis, and Francisella tularensis) when released as aerosols in the suburb of a major city. The model shows that the economic impact of a bioterrorist attack can range from an estimated $477.7 million per 100,000 persons exposed (brucellosis scenario) to $26.2 billion per 100,000 persons exposed (anthrax scenario). Rapid implementation of a postattack prophylaxis program is the single most important means of reducing these losses. By using an insurance analogy, our model provides economic justification for preparedness measures. PMID- 9204290 TI - Hantaviruses: a global disease problem. AB - Hantaviruses are carried by numerous rodent species throughout the world. In 1993, a previously unknown group of hantaviruses emerged in the United States as the cause of an acute respiratory disease now termed hantavirus pulmonary syndrome (HPS). Before than, hantaviruses were known as the etiologic agents of hemorrhagic fever with renal syndrome, a disease that occurs almost entirely in the Eastern Hemisphere. Since the discovery of the HPS-causing hantaviruses, intense investigation of the ecology and epidemiology of hantaviruses has led to the discovery of many other novel hantaviruses. Their ubiquity and potential for causing severe human illness make these viruses an important public health concern; we reviewed the distribution, ecology, disease potential, and genetic spectrum. PMID- 9204291 TI - Japanese spotted fever: report of 31 cases and review of the literature. AB - Spotted fever group (SFG) rickettsioses, which are transmitted by ticks, were long thought not to exist in Japan. Three clinical cases of Japanese spotted fever (JSF) were first reported in 1984. The causative agent was isolated and named Rickettsia japonica. Through October 1996, 31 cases were diagnosed as JSF in Tokushima Prefecture. Infected patients typically had acute high fever, headache, and characteristic exanthema; eschar was observed in 90%. After the discovery of JSF, more than a hundred cases were reported in southwestern and central Japan. Recent surveys show ticks to be the most probable vectors. As an emerging infectious disease, JSF is not commonly recognized by clinicians; therefore, even though it has not caused fatal cases, it merits careful monitoring. PMID- 9204292 TI - Polycystic kidney disease: an unrecognized emerging infectious disease? AB - Polycystic kidney disease (PKD) is one of the most common genetic diseases in humans. We contend that it may be an emerging infectious disease and/or microbial toxicosis in a vulnerable human subpopulation. Use of a differential activation protocol for the Limulus amebocyte lysate (LAL) assay showed bacterial endotoxin and fungal (1-->3)-beta-D-glucans in cyst fluids from human kidneys with PKD. Fatty acid analysis of cyst fluid confirmed the presence of 3-hydroxy fatty acids characteristic of endotoxin. Tissue and cyst fluid from three PKD patients were examined for fungal components. Serologic tests showed Fusarium, Aspergillus, and Candida antigens. IgE, but not IgG, reactive with Fusarium and Candida were also detected in cyst fluid. Fungal DNA was detected in kidney tissue and cyst fluid from these three PKD patients, but not in healthy human kidney tissue. We examine the intertwined nature of the actions of endotoxin and fungal components, sphingolipid biology in PKD, the structure of PKD gene products, infections, and integrity of gut function to establish a mechanistic hypothesis for microbial provocation of human cystic disease. Proof of this hypothesis will require identification of the microbes and microbial components involved and multifaceted studies of PKD cell biology. PMID- 9204294 TI - The rickettsia: an emerging group of pathogens in fish. AB - Piscirickettsia salmonis is the first of the previously unrecognized rickettsial pathogens of fish to be fully characterized. Since the recognition of P. salmonis in 1989, the impact of rickettsial pathogens in fish has become increasingly apparent. Growing awareness of the emergence of these fastidious intracellular organisms has led to the discovery of rickettsial diseases among diverse species of fish from different geographic locations and aquatic environments. The source, reservoir, and mode of transmission of these agents as well as appropriate methods of disease prevention and control remain to be established. PMID- 9204293 TI - Borna disease. AB - Borna disease virus, a newly classified nonsegmented negative-strand RNA virus with international distribution, infects a broad range of warm-blooded animals from birds to primates. Infection causes movement and behavioral disturbances reminiscent of some neuropsychiatric syndromes. The virus has not been clearly linked to any human disease; however, an association between infection with the virus and selected neuropsychiatric disorders has been suggested. We reviewed recent advances in Borna disease virus research, focusing on evidence of infection in humans. PMID- 9204295 TI - Rhodococcus equi and Arcanobacterium haemolyticum: two "coryneform" bacteria increasingly recognized as agents of human infection. AB - Rhodococcus equi and Arcanobacterium haemolyticum, formerly classified in the genus Corynebacterium, are members of the loosely defined taxon "coryneform" bacteria. Although they are the etiologic agents of distinct human infections, both organisms are frequently overlooked, which results in missed or delayed diagnoses. R. equi, long known as an important pathogen of immature horses, has become in the past three decades an opportunistic pathogen of severely immunosuppressed humans. Most cases are secondary to HIV infection. When specifically sought in throat swab cultures, A. haemolyticum is found responsible for 0.5% to 2.5% of bacterial pharyngitis, especially among adolescents. These two microorganisms represent a spectrum of disease in humans: from a mild, common illness to a rare life-threatening infection. Each organism elaborates lipid hydrolyzing enzymes (cholesterol oxidase by R. equi and sphingomyelinase D by A. haemolyticum) that are toxic to animals and humans and damaging to mammalian cell membranes. The participation of the cytotoxins in pathogenicity is discussed. Greater awareness of the properties of these two bacteria may promote faster, more accurate diagnoses and better clinical management. PMID- 9204296 TI - Is Creutzfeldt-Jakob disease transmitted in blood? AB - Creutzfeldt-Jakob disease (CJD) has been considered infectious since the mid 1960s, but its transmissibility through the transfusion of blood or blood products is controversial. The causative agent's novel undefined nature and resistance to standard decontamination, the absence of a screening test, and the recognition that even rare cases of transmission may be unacceptable have led to the revision of policies and procedures worldwide affecting all facets of blood product manufacturing from blood collection to transfusion. We reviewed current evidence that CJD is transmitted through blood. PMID- 9204298 TI - An unusual hantavirus outbreak in southern Argentina: person-to-person transmission? Hantavirus Pulmonary Syndrome Study Group for Patagonia. AB - Hantavirus pulmonary syndrome is a rodent-borne zoonosis first recognized in the United States in 1993. Person-to-person transmission has not been reported; however, in the outbreak of 20 cases reported here, epidemiologic evidence strongly suggests this route of transmission. PMID- 9204297 TI - A new tick-borne encephalitis-like virus infecting New England deer ticks, Ixodes dammini. AB - To determine if eastern North American Ixodes dammini, like related ticks in Eurasia, maintain tick-borne encephalitis group viruses, we analyzed ticks collected from sites where the agent of Lyme disease is zoonotic. Two viral isolates were obtained by inoculating mice with homogenates from tick salivary glands. The virus, which was described by reverse transcriptase polymerase chain reaction and direct sequencing of the amplification products, was similar to, but distinct from, Powassan virus and is provisionally named "deer tick virus." Enzootic tick-borne encephalitis group viruses accompany the agents of Lyme disease, babesiosis, and granulocytic ehrlichiosis in a Holarctic assemblage of emergent deer tick pathogens. PMID- 9204299 TI - Pertussis in The Netherlands: an outbreak despite high levels of immunization with whole-cell vaccine. AB - In 1996, a sudden increase in pertussis incidence was reported in the Netherlands (2.1 per 100,000 in 1995, 18 per 100,000 in 1996). Although not all potential surveillance artifacts could be excluded, it is highly probable that the data reflect a true outbreak. However, the cause of this increase has not yet been determined. Further research is directed to the severity of disease and a possible mismatch between the vaccine and the circulating Bordetella strains. PMID- 9204300 TI - Invasive Haemophilus influenzae type b disease in elderly nursing home residents: two related cases. AB - We investigated two fatal cases of invasive Haemophilus influenzae type b (Hib) infection in a community nursing home in western Sydney, Australia. Two elderly women had lived in the same room, and the onset of their illness was 5 days apart. Hib isolates from blood cultures showed identical profiles by pulsed field gel electrophoresis. These findings suggest that Hib infection was transmitted within this nursing home. Serious Hib disease may be underrecognized in this setting. Continued surveillance and serotyping of invasive H. influenzae disease is essential for identifying groups at increasing risk that may benefit from immunization against Hib. PMID- 9204302 TI - Gestational psittacosis in a Montana sheep rancher. AB - In humans, psittacosis is primarily a flulike illness following exposure to psittacine birds. In rare cases, pregnant women exposed to Chlamydia psittaci can contract gestational psittacosis: atypical pneumonia, sepsis, and placental insufficiency resulting in premature birth or miscarriage. In the United States, only two cases of gestational psittacosis have been reported, both from exposure to psittacine birds. Eleven other cases have been reported worldwide, mostly in the United Kingdom, all from exposure to infected birth fluids and membranes of farm mammals, notably sheep and goats. In these mammals, C. psittaci inhabit the reproductive tract, are transmitted sexually or by the fecal-oral route, and cause miscarriages. The case of gestational psittacosis in a Montana sheep rancher is the first farm animal-related case reported in the United States. Pregnant women should avoid close contact with C. psittaci-infected animals, particularly sheep and goats during the birthing season. Obstetricians should consider this diagnosis along with early antibiotic treatment and cesarean section delivery in the context of the patient's case history. PMID- 9204301 TI - Seroepidemiologic studies of hantavirus infection among wild rodents in California. AB - A total of 4,626 mammals were serologically tested for antibodies to Sin Nombre virus. All nonrodent species were antibody negative. Among wild rodents, antibody prevalence was 8.5% in murids, 1.4% in heteromyids, and < 0.1% in sciurids. Of 1,921 Peromyscus maniculatus (deer mice), 226 (11.8%) were antibody positive, including one collected in 1975. The highest antibody prevalence (71.4% of 35) was found among P. maniculatus on Santa Cruz Island, off the southern California coast. Prevalence of antibodies among deer mice trapped near sites of human cases (26.8% of 164) was significantly higher than that of mice from other sites (odds ratio = 4.5; 95% confidence interval = 1.7, 11.6). Antibody prevalence increased with rising elevation (> 1,200 meters) and correlated with a spatial cluster of hantavirus pulmonary syndrome cases in the Sierra Nevada. PMID- 9204304 TI - Rabies postexposure prophylaxis survey--Kentucky, 1994. AB - A survey of rabies postexposure prophylaxis administered by local health departments for a 1-year period showed that very few patients received treatment as a result of exposure to a confirmed rabid animal. Most prophylaxis was administered for contact with domestic animals in situations where existing recommendations for quarantine or laboratory testing of the animal were not followed. Because rabies in domestic animals in Kentucky is uncommon, these findings suggest that had the existing recommendations been followed, the prophylaxis would have been unnecessary in most cases. PMID- 9204305 TI - Biologic terrorism--responding to the threat. PMID- 9204303 TI - Lack of serologic evidence for an association between Cache Valley Virus infection and anencephaly and other neural tube defects in Texas. AB - We tested the hypothesis that Cache Valley Virus (CVV), an endemic North American bunyavirus, may be involved in the pathogenesis of human neural tube defects. This investigation followed a 1990 and 1991 south Texas outbreak of neural tube defects with a high prevalence of anencephaly and the demonstration in 1987 that in utero infection by CVV was the cause of outbreaks of central nervous system and musculoskeletal defects in North American ruminants. Sera from 74 women who gave birth to infants with neural tube defects in south Texas from 1993 through early 1995 were tested for CVV neutralizing antibody. All tested sera did not neutralize CVV. These data suggest that CVV is not involved in the induction of human neural tube defects during nonepidemic periods but do not preclude CVV involvement during epidemics. Other endemic bunyaviruses may still be involved in the pathogenesis of neural tube defects or other congenital central nervous system or musculoskeletal malformations. PMID- 9204308 TI - Global aspects of emerging and potential zoonoses: a WHO perspective. AB - Many new human pathogens that have emerged or reemerged worldwide originated from animals or from products of animal origin. Many animal species as well as categories of agents have been involved in the emergence of diseases. Wild (e.g., bats, rodents) as well as draught animals (e.g., horses) and food animals (e.g., poultry, cattle) were implicated in the epidemiologic cycles of these diseases. Many of the agents responsible for new infections and diseases in humans were viruses (e.g., hantaviruses, lyssaviruses, and morbilliviruses), but bacteria, especially enteritic bacteria (e.g., Salmonellae and Escherichia coli) and parasites (e.g., Cryptosporidium) of animal origin, were also involved in major food and waterborne outbreaks. The public health relevance of some of these agents (e.g., new lyssaviruses and morbilliviruses) is not yet fully assessed. In addition the zoonotic nature of some other human diseases, such as Ebola and the new variant form of Creutzfeldt-Jakob disease, is suspected but not yet demonstrated. Finally, the possible future use of xenografts may lead, if precautions are not taken, to the emergence of new diseases called xenozoonoses. PMID- 9204306 TI - The hantaviruses of Europe: from the bedside to the bench. AB - In Europe, hantavirus disease can hardly be called an emerging zoonosis; it is rather a rediscovered disease. Since 1934 an epidemic condition with primarily renal involvement has been described in Sweden. Nowadays, hundreds to thousands of cases per year are registered in Fennoscandia, fluctuating with the numbers of the specific Arvicoline-rodent reservoir, the red bank vole, which carries the main European serotype, Puumala (PUU). In the early 1980s, the rat-transmitted serotype, Seoul (SEO), caused laboratory outbreaks throughout Europe, and recent reports also suggest sporadic, wild rat-spread hantavirus disease. In the Balkans, at least four serotypes are present simultaneously: PUU, SEO, the "Korean" prototype Hantaan (HTN) or HTN-like types, and Dobrava, the latter causing a mortality rate of up to 20%. Moreover, recent genotyping studies have disclosed several PUU-like genotypes spread in Europe and/or Russia by other genera of the Arvicoline-rodent subfamily: Tula, Tobetsu, Khabarovsk, and Topografov. Their importance for human pathogenicity is still unclear, but serologic cross-reactions with PUU antigen might have caused their misdiagnosis as PUU-infections in the past. PMID- 9204307 TI - Brucellosis: an overview. AB - Brucellosis remains a major zoonosis worldwide. Although many countries have eradicated Brucella abortus from cattle, in some areas Brucella melitensis has emerged as a cause of infection in this species as well as in sheep and goats. Despite vaccination campaigns with the Rev 1 strain, B. melitensis remains the principal cause of human brucellosis. Brucella suis is also emerging as an agent of infection in cattle, thus extending its opportunities to infect humans. The recent isolation of distinctive strains of Brucella from marine mammals has extended its ecologic range. Molecular genetic studies have demonstrated phylogenetic affiliation to Agrobacterium, Phyllobacterium, Ochrobactrum, and Rhizobium. Polymerase chain reaction and gene probe development may provide more effective typing methods. Pathogenicity is related to production of lipopolysaccharides containing a poly N-formyl perosamine O chain, CuZn superoxide dismutase, erythrlose phosphate dehydrogenase, stress-induced proteins related to intracellular survival, and adenine and guanine monophosphate inhibitors of phagocyte functions. Protective immunity is conferred by antibody to lipopolysaccharide and T-cell-mediated macrophage activation triggered by protein antigens. Diagnosis still centers on isolation of the organism and serologic test results, especially enzyme immunoassay, which is replacing other methods. Polymerase chain reaction is also under evaluation. Therapy is based on tetracyclines with or without rifampicin, aminoglycosides, or quinolones. No satisfactory vaccines against human brucellosis are available, although attenuated purE mutants appear promising. PMID- 9204309 TI - Epidemiology of emerging zoonoses in Israel. PMID- 9204310 TI - Electronic media and emerging zoonoses. PMID- 9204311 TI - The reemergence of Aedes aegypti in Arizona. PMID- 9204312 TI - Exudative pharyngitis possibly due to Corynebacterium pseudodiphtheriticum. PMID- 9204313 TI - The Junior Radiologists Forum of the European Association of Radiology. A brief outline of its role and performed and planned activities. PMID- 9204314 TI - Organization of research in radiology: the American model. PMID- 9204315 TI - Research project design in diagnostic radiology. PMID- 9204316 TI - Ethics and research. PMID- 9204317 TI - How to present research data consistently in a scientific paper. AB - The paper analyzes, on a subjective basis, aspects of how to write scientific papers that will be accepted for publication in peer review journals. For each individual section of the manuscript, i.e. Introduction, Materials and methods, Results, Discussion, Abstract, and References, general comments and examples are given. It is concluded that writing scientific articles is a form of mental exercise that has to be practised to be successful. PMID- 9204318 TI - How to organize animal research in radiology. PMID- 9204319 TI - MR anatomy and small lesions of the eye: improved delineation with a special surface coil. AB - To develop an improved investigation protocol for MRI studies of intraocular lesions, imaging with a small surface coil (diameter 6 cm) was compared with a standard surface coil (diameter 11 cm). Both coils were assessed initially on an eye phantom and then by studying 22 patients with uveal melanoma and similar lesions of the eye. The influence of bandwidth and field or view (FOV) were systematically studied and evaluated quantitatively. A smaller bandwidth improved image quality independent of surface coil size. The subsequent secondary increase in chemical shift artefact was acceptable. Smaller FOVs (60-80 mm) necessitated the use of a smaller surface coil. A smaller bandwidth also proved to be advantageous with the use of the smaller surface coil. In conclusion, a smaller diameter surface coil improves MR imaging of ocular lesions. Pulse sequences with a small bandwidth maintain an acceptable signal-to-noise ratio when the FOV is reduced. PMID- 9204320 TI - CT and MR imaging of craniopharyngioma. AB - We reviewed imaging findings of CT and MR imaging in 20 cases of surgically confirmed craniopharyngioma in an attempt to determine their relation to patterns of tumor extent. The relationship between these patterns and the frequency of preoperative CT diagnosis and MR imaging diagnosis according to the surgical diagnosis were determined. The CT technique was superior to MR imaging in the detection of calcification. The MR imaging technique was superior to CT for determining tumor extent and provided valuable information about the relationships of the tumor to surrounding structures. Thus, CT and MR imaging have complementary roles in the diagnosis of craniopharyngiomas. In cases of possible craniopharyngioma, noncontrast sagittal T1-weighted images may enable the identification of the normal pituitary, possibly leading to the correct diagnosis. PMID- 9204321 TI - A case of bilateral reversible thalamic lesions. AB - We describe a case of bilateral reversible thalamic lesions with no neurological deficits and a good prognosis. The lesions appeared as low-density areas on CT and high-intensity areas on T2-weighted MR imaging, and resolved within 1 month, suggesting that the cause was edema. PMID- 9204322 TI - CT and MRI of amyloidoma of the CNS. AB - We present a case of cerebral amyloidoma in a 71-year-old woman. The clinical presentation and the radiological features were both highly suggestive for a brain tumor. PMID- 9204323 TI - Cervical diastematomyelia and syringohydromyelia in a myelomeningocele patient. AB - A case of cervical diastematomyelia and syringohydromyelia in a 16-year-old female myelomeningocele patient is reported. Progressive weakness of the upper extremity led to an MR examination of the brain and spine, which revealed hydrocephalus, Chiari II malformation, cervical diastematomyelia with a syringohydromyelic cavity in each hemicord and a large dural sac in the lumbar region. Operative therapy consisted of detethering and shunting of the two syringes. Soon after surgery her symptoms improved. The need for early complete MR imaging of myelomeningocele patients presenting with new symptoms is emphasized. PMID- 9204324 TI - Kissing osteochondromata leading to synostoses. AB - Our aim was to determine the incidence of synostoses in the bones of the lower limbs in patients with multiple cartilaginous exostosis (MCE) and use the available imaging to suggest the cause and mechanism of its development. Radiographs of the lower legs of 21 patients with MCE were reviewed. With the intention of demonstrating the exact site and extent of synostoses and other bone deformities, such as bone pressure atrophy or erosions in five patients, 8 proximal and 6 distal tibiofibular joints were examined by CT scans. No synostoses were present in 11 patients and 10 patients had 1 to 4 synostoses. Of these synostoses, 14 were localized below the knee joint and 9 above the ankle joint. A growing osteochondroma arising from tibia or fibula can cause an erosion in the contagious surface of the neighbouring bone. If facing osteochondromata are present in both bones and show an interlocking growth at abutting parts, on osseous fusion can take place with formation of a synostosis in the proximal or distal tibiofibular joint region. In adult patients with MCE and abundant osteochondromata synostoses between the neighbouring bones of the lower legs are common findings; they are always caused by coalescence of "kissing" osteochondromata. PMID- 9204325 TI - Bone marrow relaxation times in Gaucher disease before and after enzyme replacement therapy. AB - The aim of this work was to monitor the effectiveness of enzyme replacement therapy on the basis of the changes in T1 relaxation times in Gaucher patients. A total of 26 patients underwent MR before enzyme replacement therapy; of them, 18 have been followed-up. A total of 22 age-matched controls underwent the same MR study. Scans were focused on the femoral neck, and T1 relaxation times were measured by means of a mixed spin-echo inversion recovery sequence. The T1 relaxation times in Gaucher patients were significantly longer than normal (p < 0.05). After enzyme replacement therapy, T1 relaxation times gradually became closer to those of control subjects, and there was also a significant decrease (p < 0.01) with respect to values before therapy, probably due to an increase in the fat/water ratio. Evaluation of T1 relaxation time may supply a useful indication of Gaucher disease regression after enzyme replacement therapy particularly in those cases in which a normal skeletal appearance corresponds to prolonged T1 relaxation times. PMID- 9204326 TI - Imaging findings of sternal abnormalities. AB - Radiographic findings in the sternal abnormalities are often nonspecific, showing appearances from a localized benign lesion to an aggressive lesion as seen with infections and malignant neoplasms. A specific diagnosis of sternal abnormalities can be suggested on the basis of CT and MR characteristics. Familiarity with the presentation and variable appearance of sternal abnormalities may aid the radiologist is suggesting a specific diagnosis. We present among others characteristic radiographic findings of hemangioma, chondrosarcoma, hydatid disease, and SAPHO syndrome. In those cases in which findings are not specific, cross-sectional imaging modalities may help the clinician in their management. PMID- 9204328 TI - Solitary plasmacytomas of the occipital bone: a report of two cases. AB - The radiological appearances of two cases of solitary plasmacytoma in the occipital bone are described. One arose in the lateral part and the other in the squama. They showed characteristic radiological features on CT, MRI and angiography. Bone scintigraphy and gallium scintigraphy were also available. Solitary plasmacytoma of the skull is a rare condition and usually occurs in the calvarium. The skull base is an extremely rare site and only four cases have been reported. The literature of solitary plasmacytoma of the skull is reviewed. PMID- 9204327 TI - Contrast media in MR arthrography of the glenohumeral joint: intra-articular gadopentetate vs saline: preliminary results. AB - The purpose of this investigation was to compare gadopentetate and saline as contrast media in MR arthrograms of the glenohumeral joint. In 60 consecutive patients MR arthrograms with either gadopentetate (n = 26) or saline (n = 34) were performed. After injection of gadopentate, 3D gradient-echo (GE) images were obtained (TR 32 ms, TE 10 ms, flip angle 40 degrees). With saline, double-echo steady-state images (heavily T2-weighted 3D GE images) were obtained (TR 40 ms, TE 9/45 ms, flip angle 40 degrees). In the last 14 of these patients T2-weighted turbo spin-echo (SE) images were added (TR 2900 ms, TE 96 ms). Contrast-to-noise ratios standardized for imaging times proved to be superior for the gadolinium arthrograms compared with GE and SE saline arthrograms (intra-articular fluid vs subacromial fat: p = 0.0001 and 0.0008; intra-articular fluid vs supraspinatus tendon: p = 0.0001 and 0.046). Using a qualitative scoring system gadolinium arthrograms were superior to saline arthrograms (p < 0.0001 and p < 0.0001). Saline arthrograms in combination with GE and SE sequences are inferior to gadopentetate arthrograms with GE sequences. PMID- 9204329 TI - Contrast agents for MR imaging of the liver: a clinical overview. AB - Different contrast agents have been clinically used in MR imaging of the liver including extracellular gadolinium chelates, contrast agents targeted to the macrophage-monocytic phagocytic system (MMPS), hepatobiliary contrast agents, and blood-pool contrast agents. Extracellular gadolinium chelates are optimally used for characterization of focal hepatic lesions, whereas hepatobiliary and MMPS targeted contrast agents are optimally used for detection and preoperative evaluation. The present review portrays these contrast agents and discusses their advantages and shortcomings. PMID- 9204330 TI - Long-term results of percutaneous ethanol injection therapy for hepatocellular carcinoma in cirrhosis: a European experience. AB - The objective of our work was to evaluate the long-term results of percutaneous ethanol injection (PEI) for the treatment of hepatocellular carcinoma (HCC) in patients with liver cirrhosis. A total of 184 cirrhotic patients with HCC underwent PEI as the only anticancer treatment over an 8-year period. Patients were followed after therapy by means of clinical examinations, laboratory tests, and US and CT studies performed at regular time intervals. Survival rates were determined according to the Kaplan-Meier method. The overall survival was 67% at 3 years, 41% at 5 years, and 19% at 7 years. The 3-, 5-, and 7-year survival rates of patients with single HCC < or = 3 cm (78, 54, and 28%, respectively) were significantly higher (p < 0.01) than those of patients with single HCC of 3.1-5 cm (61, 32, and 16, respectively) or multiple HCCs (51, 21, and 0%, respectively). Survival of Child-Pugh A patients (79% at 3 years, 53% at 5 years, and 32% at 7 years) was significantly longer (p < 0.01) than that of Child-Pugh B patients (50% at 3 years, 28% at 5 years, and 8% at 7 years). A selected group of 70 patients with Child-Pugh A cirrhosis and single HCC < or = 3 cm had a 7-year survival of 42%. Long-term survival of cirrhotic patients with HCC treated with PEI is comparable to that reported in published series of matched patients submitted to surgical resection. PMID- 9204331 TI - Choledocholithiasis after Billroth II surgery: MR cholangiographic diagnosis. AB - In this case of choledocholithiasis in a patient with previous Billroth-II surgery and cholecystectomy we demonstrate the advantages of a heavily T2 weighted half-Fourier turbo-spin-echo (HASTE) sequence. This technique allows thin-slice snapshot imaging with 1.4 s per slice eliminating motion artifacts and still has the necessary heavy T2-weighting to depict biliary fluid with high contrast. In the presented case endoscopic retrograde cholangiography (ERCP) could not be performed prior to MRI due to technical problems. In a second attempt, ERCP was successful and a common bile duct stone as diagnosed by MRI before could be removed. We conclude that HASTE snapshot MR cholangiography can be used as a clinically valuable tool when other noninvasive methods are not diagnostic. PMID- 9204332 TI - Posttraumatic intestinal stenosis: radiographic and sonographic appearance. AB - We report a case of posttraumatic intestinal stenosis (PIS), an uncommon sequela of blunt abdominal trauma, in which injury to the mesentery and bowel wall results in later focal ischemic stricture of that segment. We include CT images at the time of trauma, and barium meal and abdominal sonography obtained during the subsequent admission. Examination of the resected bowel loop showed transmural infarct and posttraumatic changes in the adjacent peritoneal fat. This is the first report which includes both imaging at the time of trauma and sonographic appearance of the narrowed bowel loop. Posttraumatic intestinal stenosis should be considered in the differential diagnosis of a narrowed bowel loop in a patient with a history of blunt abdominal trauma. PMID- 9204334 TI - Is it possible to choose the safest contrast media through the results of clinical studies? PMID- 9204335 TI - Quality assurance programme applied to mobile C-arm fluoroscopy systems. AB - The importance of quality assurance (QA) of X-ray equipment in diagnostic imaging departments is well recognised. However, practically no attention has been paid in the literature to the application of QA programmes to mobile C-arm fluoroscopy systems. This equipment is sometimes omitted from these programmes because it is often "off-site" from the main radiological facility and suitable QA protocols are unavailable. The need for QA can be substantiated by the fact that these systems are finding greater clinical use in orthopaedic, vascular and cardiac applications. Hence, there is a growing awareness among users for the need of good image quality and low patient radiation dose. In view of this, the objective of this study was to review the existing literature, design a suitable QA protocol for this equipment and use it to survey 10 C-arms in clinical use. The protocol was designed to address mechanical and electrical safety in addition to radiation safety and image quality. Results indicate substantial performance differences between systems with significant variations in input air kerma rate to the image receptor. The authors believe that such a protocol is necessary with a view to establishing optimal performance levels and assist in the development of suitable "write-off" criteria for such systems. PMID- 9204333 TI - Application of a stable cell culture assay for the functional assessment of novel MR contrast agents. AB - The purpose of this study was to apply a new cell culture assay that preserves hepatocyte orientation and differentiation for screening of MR contrast agents with hepatocyte specificity. Cultured hepatocytes were sandwiched between two layers of collagen, preserving both hepatocyte function and morphology over a prolonged period of time. Plain and rhodaminated monocrystalline iron-oxide particles (MION and MION-rh) and asialoglycoprotein receptor-specific rhodaminated asialofetuin coupled to MION (MION-ASF-rh) were prepared. Dose dependent competition experiments of these agents were performed with D(+) galactose to determine the specificity of galactose-mediated cell uptake. To assess the impact of cell integrity on cell uptake dose-dependent functional experiments with two hepatotoxins (ethanol and CCl4) were performed. Normal cell cultures showed significantly higher fluorescent-light emission after incubation with hepatocyte-directed ASF-MION-rh than after incubation with MION-rh. Competition experiments of ASF-MION with galactose showed a dose-dependent decrease in calibrated fluorescent-light emission. Cell cultures treated with hepatotoxins demonstrated a dose-dependent reduction in calibrated fluorescent light emission following incubation with ASF-MION-rh. The validated assay system allows assessment not only of hepatocyte specificity, but also of hepatocyte damage. Because the assay can be applied to cells from any species (rat, pig, human), it may represent an ideal test system prior to clinical trials of new hepatocyte-directed MR contrast agents. PMID- 9204336 TI - Diagnostic value of MR imaging in comparison to CT in the detection and differential diagnosis of renal masses: ROC analysis. AB - The aim of this work was to compare MR imaging and CT in the detection of renal masses and in the differential diagnosis between benign and malignant lesions. In 33 patients with 54 renal lesions CT and MR images were evaluated by four readers with regard to tumor detection and characterization using a receiver-operating characteristics (ROC) analysis. The MRI protocol consisted of a T1-weighted spin echo (SE) sequence (TR/TE: 300/10 ms) before and after contrast administration and a heavily T2-weighted turbo-SE (TSE) sequence (TR/TE: 5500/150 ms). Az values for the area under the ROC curves for lesion detection were 0.92 +/- 0.04 for CT and 0.91 +/- 0.05 for MRI, respectively, which was not statistically different. The MRI technique was slightly, but not significantly, better than CT in the overall characterization (accuracy in differentiation between benign and malignant) of renal lesions with an Az value of 0.90 +/- 0.05 compared with 0.88 +/- 0.06 for CT. The MRI technique proved to be statistically superior to CT (p < 0.01) in the correct characterization of benign renal lesions. MRI equals CT in the overall detection and differential diagnosis of renal masses. MRI is very helpful for further differential diagnosis of lesions which are equivocal on CT especially in the differentiation between complicated cysts and cystic or hypovascular renal cell carcinoma. PMID- 9204338 TI - Amyloidosis of the seminal vesicles simulating tumor invasion of prostatic carcinoma on endorectal MR images. AB - Amyloid deposits within the seminal vesicles are a common finding at autopsy. The incidence increases with age. Amyloid deposits can mimic tumor extension into the seminal vesicles due to prostate or bladder cancer on T2-weighted MR images. We describe a case of seminal vesicle amyloidosis demonstrating the MR appearance and the characteristic pathologic findings. Recognizing seminal vesicle amyloidosis may prevent overstaging prostate cancer on MR images. PMID- 9204337 TI - Imaging findings in renal hydatid disease. AB - The aim of this work is to describe the image findings of renal hydatid disease, especially on MR. Four cases of echinococcal involvement of the kidney were retrospectively reviewed. All patients had intravenous urography (IVU) and US performed. Computed tomography examination was available in three patients and MR in two cases. Intravenous urography demonstrated communication of the cyst to the collecting system in one case. Ultrasound revealed multicystic appearance in three cases and unilocular in one case. Computed tomography demonstrated unilocular thick-walled or multilocular cysts with well-defined walls, calcified in one case. In multilocular cysts the CT densities of the fluid of daughter cysts was significantly lower than the fluid of mother cysts. This typical appearance was present in three of our cases. The presence of a hypointense rim and a multicystic appearance were distinctive in MR imaging. The combined findings of these different imaging modalities aid greatly in establishing the correct diagnosis. Magnetic resonance imaging is of value in determining the presence of a characteristic rim and enables the evaluation of anatomical relationships. PMID- 9204339 TI - MRI, CT, and TRUS imaging of seminal vesicle metastasis. PMID- 9204340 TI - Late presentation of solitary contralateral adrenal metastasis of renal cell carcinoma. AB - We report a case of renal cell carcinoma with solitary metachronous contralateral adrenal metastasis occurring 23 years after radical nephrectomy. The patient was treated with adrenalectomy. He is alive with no evidence of disease 1 year postoperatively. Solitary metachronous contralateral adrenal metastases from renal cell carcinoma are rare clinical conditions that may occur very late. Metastasectomy is advocated and is probably beneficial for limited metastatic renal cell cancer. PMID- 9204341 TI - Ultrasound of the hollow gastrointestinal tract in children. PMID- 9204342 TI - MRI of the nasal cavity, the paranasal sinuses and orbits in Wegener's granulomatosis. AB - The purpose of this study was to evaluate diagnostic MRI criteria in Wegener's granulomatosis of the nasal cavity, the paranasal sinuses and orbits. Between March 1991 and January 1996, 62 patients with biopsy-proven Wegener's granulomatosis were studied with T1- and T2-weighted spin-echo (SE) sequences. In 32 patients coronal postcontrast T1-weighted images were obtained. Mucosal thickening of the nasal cavity and paranasal sinuses was demonstrated as high intensity lesions on T2-weighted SE sequences in 57 patients (92%). Of this group, inflammatory granulomatous tissue was found on biopsy in 30 patients (48%) in the nasal cavity and in 4 patients (6%) in the paranasal sinuses. In 23 patients (37%) biopsy revealed unspecific inflammatory changes without evidence of granulomatous tissue. In 14 patients (23%) granulomas were depicted as low signal intensity lesions on T1- and T2-weighted SE sequences in the paranasal sinuses and orbits. In 5 patients (8%) osseous destruction was found. After gadolinium injection, 12 of 14 granulomas showed inhomogeneous signal enhancement. In two granulomas no enhancement was found. The MRI technique is helpful in the diagnosis of patients with Wegener's granulomatosis. In the initial inflammatory process of Wegener's granulomatosis, it is not possible to differentiate between mucosal inflammation and granulomatous tissue in MRI. In the later stage of granulomatous transformation, granulomas can be depicted as low-signal-intensity lesions. Therefore, Wegener's granulomatosis should be included in the differential diagnosis of patients with low-signal-intensity lesions on T1- and T2-weighted SE sequences of the nasal cavity, paranasal sinuses and orbits. PMID- 9204343 TI - Antrochoanal autopolypectomy: CT findings. AB - Antrochoanal polyp (Killian polyp) is an infrequent, benign lesion of maxillary origin. We describe the basic characteristics of this lesion and a rare case of autopolypectomy. Coronal and axial CT images are presented before and after autoexpulsion of an antrochoanal polyp in a patient with long-standing nasal obstruction. The initial CT examination revealed a typical left antrochoanal polyp filling all the maxillary sinus and passing through the ethmoid infundibulum until the choana. The CT after autopolypectomy showed the secondary mass effect at surrounding structures and residual inflammatory changes. PMID- 9204344 TI - Eccrine porocarcinoma with intracerebral extension. PMID- 9204346 TI - Pneumomediastinum and pneumopericardium due to malignant subcarinal lymphadenopathy: CT demonstration. AB - A 52-year-old man had been treated for oral cancer T3 N0 M0 by radical surgery, neck dissection on the right and cervical irradiation (60 Gy). Two months after therapy he presented with dysphagia and hemoptysis. Admission chest X-ray revealed a pneumopericardium. It was caused by a bronchomediastinal fistula due to necrotic metastatic lymph nodes as shown by CT, which also revealed a concomitant pneumomediastinum. The patient died 10 days later from pneumonia. The CT findings were confirmed at autopsy. We conclude that malignant mediastinal lymphadenopathy is a potential cause of pneumopericardium and pneumomediastinum. PMID- 9204345 TI - Detection of radiographic findings in lung fibrosis using storage phosphor plates. AB - The aim of this work was to examine the influence of the filter kernel size on the detectability of differing radiological findings in interstitial lung disease. In 97 patients with confirmed pulmonary fibrosis chest radiographs were obtained with a filmscreen system of speed class 200 and with correspondingly exposed storage phosphorous plates. The size of the filter kernel used for the image postprocessing varied between sigma 5 and sigma 70. The detectability of interstitial lung changes was evaluated independently by eight readers on the basis of a defined rating system. The results were analysed using multifactorial analysis of variance with Scheffe test at a significance level of p = 0.05. Small kernel sizes (S 5, S 10) combined with high edge enhancement were only of benefit in the imaging of septal lines, but reduced the detectability of nodular and reticular structures. Good detail detectability of both micronodules and septal lines was obtained with a medium kernel size of sigma 40. Storage phosphor radiography utilizing the appropriate choice of postprocessing parameters provides equivalent image quality for evaluating interstitial lung changes compared with a modern filmscreen technique. PMID- 9204348 TI - Quiz case of the month. Multiple hamartomas of the liver. PMID- 9204347 TI - Primary solitary amyloidoma of the lung: findings on CT and MRI. AB - Pulmonary amyloidoma is a rare disease which is usually found incidentally on chest radiographs in asymptomatic, elderly people. Amyloid nodules may be solitary or much more commonly multiple. There have been many reports of radiological findings of pulmonary amyloidosis; however, those have not been characteristic. We report the findings on CT and MRI of a proven primary pulmonary amyloidoma in an asymptomatic 76-year-old woman. The low intensity of the lesion on T2-weighted images may be useful in the differential diagnosis from bronchogenic carcinoma. PMID- 9204349 TI - Iopentol (Imagopaque) in vascular procedures. A multi-centre monitoring trial assessing adverse events and diagnostic information--results from 3,587 patients in Germany. AB - The aims of the present open, non-comparative survey were to study the safety and efficacy of iopentol (Imagopaque, Nycomed Imaging AS, Oslo, Norway) in a large patient group. In a series of German centres, 3,587 patients underwent various contrast-enhanced examinations with iopentol. The most frequent examinations were computed tomography (CT) (1,740), phlebography (462), and digital subtraction angiography (DSA) (493). Only 82 patients (2.3%) experienced one or more adverse events. Sixty-one (1.7%) of these events were possibly or probably caused by the contrast medium. A total of 111 adverse events were registered, 54 of mild, 42 of moderate and 12 of strong intensity, and 51 events required treatment. The most frequent adverse events were nausea (34), erythema (14) urticaria (9), taste sensation (6), circulatory reactions (5) and angina pectoris (5). The frequencies of adverse events were 2.9% in CT, 2.0% in DSA, 2.0% in phlebography, 1.6% in cardioangiography, and 0.4% in urography. Patients with arteriosclerosis, an earlier contrast medium reaction, multimorbidity or age over 70 years had a statistically significantly higher risk of experiencing an adverse event. Patient tolerance was very good; the mean score was 83% on a visual analogue scale (VAS) ranging from extremely bad (0%) to extremely good (100%). Efficacy, as measured on VAS, was determined. Technical quality was scored as 80%, contrast enhancement within the vessels as 80% and delineation of lesions as 79%. The results from this large patient population confirms the experience from clinical practice that iopentol is a safe, well tolerated and efficient contrast medium. PMID- 9204350 TI - Evaluation of iopentol (Imagopaque 350) in CT enhancement. A multi-centre monitoring trial assessing adverse events and diagnostic information--results from 1,823 patients in France. AB - OBJECTIVES: To evaluate the safety and efficacy of iopentol 350 mg I/ml (Imagopaque/Ivepaque, Nycomed Imaging AS, Oslo, Norway), a monomeric non-ionic contrast medium for computed tomography, in a large population. To identify predictive factors for patient safety. MATERIALS AND METHODS: One thousand eight hundred and twenty-three (1,823) patients from 48 centres in France were included during a 5-month period. Safety was evaluated by registering adverse events (AEs) reported by the patients, and data were analysed using a multiple factor model. RESULTS: Only 2.6% of the patients experienced AEs other than discomfort. There were no serious AEs. Overall, AEs were more frequent in patients under 50 years of age, in women, and in patients who received contrast medium as a single bolus. Contrast enhancement was considered adequate or better in 98.9% of the patients. A large variation in discomfort (local warmth/chill or pain) frequency was seen between centres, ranging from 0% to 81%. This result implies that factors other than the CM influence the incidence of discomfort. CONCLUSIONS: This first study in a large population shows that iopentol 350 mg I/ml is well tolerated and provides CT images of excellent, good or adequate quality in the vast majority of patients. Age, sex and injection procedure were shown to be independent predictors in the AE survey. PMID- 9204351 TI - Iopentol (Imagopaque 300) compared with iopromide (Ultravist 300) in abdominal CT. A multi-centre monitoring trial assessing adverse events and diagnostic information--results from 518 patients in Spain. AB - OBJECTIVES: Iopentol (Nycomed Imaging AS, Oslo, Norway) and iopromide (Schering AG, Berlin, Germany) are low-osmolar, non-ionic, iodinated contrast media (CM) used in abdominal CT examinations. The intravenous safety profile and radiological efficacy of iopentol and iopromide were studied in 518 patients. Specifically, frequency of adverse events (AEs), subjective change in quality of diagnostic information, and quantitative enhancement characteristics were compared. MATERIALS AND METHODS: A prospective, double-blind, randomized, multicentre, parallel-group study was conducted at 8 hospitals. Patients received 100 ml of either iopentol 300 mg I/ml or iopromide 300 mg I/ml. RESULTS: The incidence of patients with AEs was statistically significantly lower in the iopentol group compared to the iopromide group (2.3% vs. 8.9%, p < 0.001). Discomfort was frequent in both groups (44.8% vs. 49.4%, p = 0.33), sensation of heat and warmth being most common. Overall, diagnostic information was similar in both groups. Both CM gave high percentages of examinations rated as optimal (87.1% vs. 90.5%, p = 0.34) and in which diagnostic confidence was increased (87.5% vs. 91.1%, p = 0.22). No significant differences between the two CM were found concerning quantitative enhancement characteristics. CONCLUSIONS: In this study iopentol was significantly safer than iopromide for contrast enhanced CT examination of the abdomen. Radiological efficacy was similar with both CM. PMID- 9204352 TI - Iopentol (Imagopaque 300) compared with iohexol (Omnipaque 300) and diatrizoate (Urografin 292) in pediatric urography. A clinical trial assessing adverse events and diagnostic information. AB - A double-blind, parallel-group trial was performed in 96 children in order to evaluate and compare the safety and efficacy of iopentol (Imagopaque, Nycomed imaging AS, Oslo, Norway) with those of routinely used contrast media. Ten children below 1 year of age received iopentol and eight received iohexol (Omnipaque, Nycomed Imaging AS, Oslo, Norway) (both 300 mg I/ml) in a random manner. Seventy-eight children (39 in each group), between one and 10 years of age, received iopentol 300 or diatrizoate (Urografin, Schering AG, Berlin, Germany) 292 mg I/ml for urography. No adverse events were observed among the children below 1 year of age. In the group between one and 10 years there was a statistically and clinically significant difference (p = 0.007) in the incidence of adverse events between the iopentol (2.6%) and diatrizoate (25.6%) groups. All adverse events were of mild or moderate intensity. No serious reactions were encountered. The overall quality of visualization was judged to be diagnostic for all patients. This study supports the use of non-ionic contrast media in pediatric patients and confirms that iopentol is a safe and effective contrast medium well suited for children from 0-10 years of age. PMID- 9204353 TI - Iopentol (Imagopaque 350) compared with iohexol (Omnipaque 350) in pediatric cardioangiography. A clinical trial assessing adverse events, ECG, blood pressure and diagnostic information. AB - A non-ionic, monomeric, low-osmolar contrast medium, iopentol (Imagopaque, Nycomed Imaging AS, Oslo, Norway) has been compared with iohexol (Omnipaque, Nycomed Imaging AS, Oslo, Norway) in a double-blind, randomized, parallel-group trial in children. The aims of the study were to compare the safety and radiographic efficacy of iopentol 350 mg I/ml versus those of iohexol 350 mg I/ ml in pediatric cardioangiography. Seventy-three children, of whom 35 received iopentol and 38 iohexol, were included in the evaluation. The quality of overall diagnostic information was excellent or adequate in all patients. There were no statistically significant differences between numbers of patients reporting adverse events nor number of patients reporting discomfort in the two groups. Three patients in the iopentol group and two patients in the iohexol group reported adverse events. Heat sensation was reported by 4 patients in the iopentol group and 5 patients in the iohexol group. Following injection of contrast medium, there were no differences between the groups regarding ECG, heart rate and systolic blood pressure at the site of injection. Iopentol was demonstrated to be as effective, safe and well tolerated as iohexol in pediatric cardioangiography. PMID- 9204354 TI - Iopentol (Imagopaque 300) compared with iopromide (Ultravist 300) in pediatric angiocardiography. A clinical trial assessing adverse events, ECG and diagnostic information. AB - Iopentol (Imagopaque, Nycomed Imaging AS, Oslo, Norway) 300 mg.l/ml was compared with iopromide (Ultravist, Schering AG. Berlin, Germany) 300 mg I/ml in pediatric angiocardiography in 97 children (48 and 49 patients, mean age 2.8 and 4.1 years in the respective contrast medium groups). The volume injected was usually 4-6 ml/kg b.w. ECG, blood pressure, heart rate, adverse events and efficacy were evaluated. Five patients in the iopentol group and four in the iopromide group reported adverse events. This difference was not statistically significant. One adverse event in each of the two groups was considered as possibly related to the contrast medium. Only small and transient changes in heart rate, blood pressure and ECG parameters were observed. No difference between the two contrast media was found for efficacy. It can be concluded that iopentol is well suited for angiocardiographic examinations in children. PMID- 9204355 TI - Iopentol (Imagopaque 250) compared with diatrizoate (Urografin 219) in endoscopic retrograde cholangio-pancreatography (ERCP). A clinical trial assessing safety (adverse events and S-pancreatic iso-amylase) and diagnostic information (VAS). AB - The efficacy and safety of the non-ionic contrast medium iopentol, 250 mg I/ml (Imagopaque, Nycomed Imaging AS, Oslo, Norway) were evaluated and compared to those of the ionic contrast medium diatrizoate 219 mg I/ml (Urografin, Schering AG, Berlin, Germany). The trial was carried out as a randomized, double-blind comparative two-group study. One hundred and sixteen patients completed the study: 59 received iopentol and 57 received diatrizoate, the contrast medium used according to routine hospital procedure for ERCP. Demographic data and details of the ERCP procedure were comparable for the two contrast medium groups. Safety was assessed by monitoring serum pancreatic iso-amylase and by recording adverse events. Efficacy was evaluated through assessment of the diagnostic information and the quality of radiographs. Twenty-one of the patients receiving iopentol and 27 of the patients receiving diatrizoate reported adverse events during the ERCP procedure. Pain was the kind of procedure-related event most frequently reported. Three patients experienced serious adverse events with a fatal outcome 2, 10 and 12 days after the ERCP procedure, respectively. The principal investigator concluded for all three serious adverse events that any causal relationship with the contrast medium injected was unlikely. A mean change in serum pancreatic iso amylase after contrast medium injection was noted in both contrast medium groups. However, statistical analysis did not show any significant difference between mean changes for the two groups. The efficacy results, both in terms of diagnostic information and quality of radiographic visualisation, were comparable for the two contrast media. In conclusion, the study did not show any statistically significant differences between the two contrast media as regards safety or efficacy in ERCP. PMID- 9204356 TI - Iopentol (Imagopaque 300) compared with ioxaglate (Hexabrix 320) in knee arthrography. A clinical trial assessing immediate and late adverse events and diagnostic information. AB - OBJECTIVES: This trial was designed to compare the incidence of delayed postprocedural pain after administration of iopentol (Imagopaque, Nycomed Imaging AS, Oslo, Norway) 300 mg I/ml versus ioxaglate (Hexabrix, Guerbet, Aulnay-sous Bois, France) 320 mg I/ml in single-contrast knee arthrography. Other adverse events and radiographic efficacy were also evaluated. METHODS: A randomized, double-blind study was performed in 120 patients. Immediate adverse events were recorded up to 30 min post-injection, and late adverse events, including knee pain and swelling, up to 4 days after the examination (by using a patient questionnaire). Diagnostic information obtained up to 20 min after injection was assessed using a Visual Analogue Scale (VAS). RESULTS: Significantly more patients in the ioxaglate group (50%) than in the iopentol group (30%) reported increased knee pain during the 4 days following the examination (p < 0.05). Adverse events up to 30 min after the examination, and late adverse events other than knee pain, were reported by similar numbers of patients in the two contrast medium groups. The diagnostic information was similar in both groups. CONCLUSION: Both iopentol and ioxaglate are effective and safe contrast media for use in knee arthrography, but iopentol induces less postprocedural pain. PMID- 9204357 TI - Oral administration of iopentol (Imagopaque 300 mg I/ml) compared with amidotrizoate (Peritrast 300 mg I/ml), both diluted to 2% (v/v), in imaging of the gastrointestinal tract in abdominal contrast enhanced CT. A clinical trial assessing patient tolerance, distribution of contrast medium and Hounsfield unit measurements. AB - The aim of the trial was to evaluate and compare the safety and efficacy of iopentol (Imagopaque, Nycomed Imaging AS, Oslo, Norway) and amidotrizoate (Peritrast, Kohler Pharma, Alsbach, Germany), both 300 mg I/ml initially, but diluted to 2% (v/v) and administered orally. Sixty-four and 65 patients were included in the respective contrast medium groups. Portions of contrast medium, totally 1.51, were taken every 15 min during the hour before the examination. A standard radiological procedure for abdominal CT was followed. Nine percent of the patients in each group experienced adverse events which were possibly contrast medium related. Taste acceptance was comparable in the two groups. Except for the stomach, radiographic efficacy was satisfactory for all intestinal segments. The difference in density in the proximal small bowel (main parameter) achieved with the media was not significant (p = 0.33), nor was that as regards image homogeneity (contrast distribution). In conclusion, iopentol is well suited for oral contrast enhancement of the gastrointestinal tract in abdominal computed tomography. PMID- 9204358 TI - Iopentol (Imagopaque 350) compared with diatrizoate (Urografin 370) in cerebral CT. A clinical trial assessing immediate and late (7 days) adverse events and diagnostic information (visualization quality and Hounsfield unit measurements). AB - The non-ionic contrast medium iopentol (Imagopaque, Nycomed Imaging AS, Oslo, Norway) 350 mg I/ml was compared for safety and efficacy with the ionic contrast medium diatrizoate (Urografin, Schering AG, Berlin, Germany) 370 mg I/ml in a randomized, double-blind, parallel-group clinical trial in cerebral computed tomography (CT). The numbers of participating patients was 79; forty in the iopentol group and 39 in the diatrizoate group. Safety was evaluated by assessing the numbers of patients reporting immediate adverse events (up to 30 min after injection) and delayed adverse events (within 7 days after the examination). Efficacy was expressed as the quality of visualization of the cerebral lesions after injection of the contrast medium. In addition, Hounsfield units were measured pre- and post-contrast. No patient in either group experienced any serious adverse event. The frequency of patients with immediate adverse events was statistically significantly lower in the iopentol group (23%) than in the diatrizoate group (64%), p = 0.0003. Delayed adverse events were also significantly less frequent in the iopentol group (43%) than in the diatrizoate group (69%), p = 0.047. Patients in the iopentol group reported significantly less discomfort (53%), especially sensation of warmth, than patients in the diatrizoate group (92%), p = 0.0001. The intensity of adverse events and injection-associated discomfort seemed, in general, to be lower for patients in the iopentol group. No difference was found between the two contrast media regarding efficacy. PMID- 9204359 TI - Delayed CT of the kidneys after iopentol (Imagopaque 350) injection in patients with normal renal function. An assessment of the gradual decrease in opacification over 32 h after injection. AB - The elimination of the non-ionic contrast medium iopentol (Imagopaque, Nycomed Imaging AS, Oslo, Norway) from the kidneys was investigated in adult patients with normal renal function referred for cerebral computed tomography (CT). Twenty four patients were included in the study. Each patient was given one intravenous injection of 50 ml iopentol 350 mg I/ml. CT scans of the kidneys were taken before, immediately after, 8 h after and 32 h after the injection of contrast medium. To assess the rate of iopentol disappearance, calculations were made for the cortex, medulla and aorta, based on changes from the baseline (pre-contrast) measurements, in Hounsfield units (HU). The median estimated time for disappearance was 28 h from the cortex, 32 h from the medulla and 8 h from the aorta. Thus, a small retention of contrast medium can usually be seen after normalization of aortic HU values. PMID- 9204360 TI - Iopentol (Imagopaque 300 and 350) compared with iohexol (Omnipaque 300 and 350) in cerebral and aortic arch angiography. A clinical trial assessing adverse events and diagnostic information. AB - The safety of the non-ionic contrast medium iopentol (Imagopaque, Nycomed Imaging AS, Oslo, Norway) when used in cerebral angiography and aortic arch angiography, was the focus of this investigation. Overall quality of visualization and changes in heart rate and blood pressure were, however, also assessed. In total, 39 patients were injected with iopentol and 41 patients with the comparative contrast medium, iohexol (Omnipaque, Nycomed Imaging AS, Oslo, Norway). Two patients (5%) in each group reported contrast-related adverse events other than a sensation of heat, while three patients in the iopentol group and four in the iohexol group reported procedure-related adverse events. A sensation of heat was reported by 21 patients (54%) in the iopentol group, and by 20 patients (49%) in the iohexol group. There were no clinically relevant changes in heart rate or blood pressure. The diagnostic information obtained was of sufficient or excellent quality for all patients. Statistical analyses did not indicate any significant difference between the two contrast media. Iopentol was well suited for cerebral and aortic arch angiography, comparable to iohexol regarding safety and efficacy. PMID- 9204362 TI - Purification and structural characterization of insulin from the lesser siren, Siren intermedia (Amphibia: Caudata). AB - Insulin has been isolated from an extract of the pancreas of a salamander, the lesser siren Siren intermedia, and its primary structure was established as: A chain, Gly-Ile-Val- Glu-Gln-Cys-Cys-His-Asn-Thr10-Cys-Ser-Leu-Tyr-Gln-Leu-Glu-Asn Tyr- Cys20-Asn, and B-chain, Val-Pro-Asn-Lys-Pro- Leu-Cys-Gly-Ala-His10-Leu-Val Glu-Val-Met-Tyr-Phe-Val- Cys-Gly20-Asp-Arg-Gly-Phe-Phe-Tyr-Pro-Ser-Ser-Thr 30. Although those amino acid residues considered to constitute the receptor-binding region of insulin have been retained, siren insulin contains several substitutions (Gln-->Lys at B4, Ser-->Ala at B9, Ala-->Val at B14, Leu-->Met at B15, Leu-->Phe at B17, Pro-->Ser at B28, and Lys-->Ser at B29) of amino acid residues that are conserved in insulins from species of other amphibian orders. The biological activity of siren insulin was not investigated in this study but the substitutions at B28 (involved in dimer formation) and at B14 and B17 (involved in hexamer formation) may be expected to influence conformation and therefore biological potency. The data are consistent with the view that the Sirenoidea represent an early divergence from the ancestral stock of salamanders. PMID- 9204361 TI - Cardiovascular and electrocardiographic effects of iopentol in left ventricular angiography. Comparison of the low-osmolar, non-ionic iopentol (Imagopaque 350) and the hyper-osmolar, ionic metrizoate meglumine-Na-Ca (Isopaque Coronar 370) in patients with coronary heart disease. AB - The aim of the study was to evaluate and compare the hemodynamic and electrocardiographic effects following injection of the non-ionic, low-osmolar contrast medium iopentol (Imagopaque 350, Nycomed Imaging AS, Oslo, Norway) and the ionic, hyper-osmolar contrast medium metrizoate meglumine-Na-Ca (Isopaque Coronar 370, Nycomed Imaging AS, Oslo, Norway) when used for left ventricular angiography. The study was performed in a double-blind, randomized manner in 82 patients with severe coronary heart disease. The patients who received iopentol experienced less adverse events and subjective discomfort of lesser intensity than those who received metrizoate (p = 0.0001). Both contrast media induced a biphasic change in left ventricular (LV) systolic pressure, with an initial fall followed by a prolonged rise, but the alterations were statistically significantly more pronounced with metrizoate than with iopentol. The changes in LV end-diastolic pressure (p = 0.023), and LV negative dP/dt (p = 0.002) were significantly more pronounced with metrizoate than with iopentol. Cardiac output and heart rate increased more with metrizoate, while stroke volume was equally increased by both agents. A prolonged increase in the QT-interval, throughout the 10-min observation period, was seen only after injection of metrizoate (p = 0.0006 for comparison between contrast media). CONCLUSION: Iopentol was well tolerated and induced markedly less severe hemodynamic and electrocardiographic alterations than did metrizoate in patients with severe coronary heart disease. PMID- 9204363 TI - Double-label immunofluorescence study of glutamic acid decarboxylase in the fetal and adult ovine pancreas by light and confocal microscopy: evidence for predominant beta-cell coexpression. AB - Glutamic acid decarboxylase (GAD) is present in the central nervous system and in several nonneuronal tissues including the pancreatic islets. There are two isoforms with molecular weights of 65 kDa (GAD65) and 67 kDa (GAD67). The cellular specificity of the two molecular forms of GAD and their levels within the mammalian islets may be species-dependent, being coexpressed in both beta and in non-beta cells. We have examined the ovine pancreas, from the adult and fetal stages of late gestation, for the expression of GAD65 within the islet cells by double-label immunofluorescence light and confocal microscopy. In the adult tissue, GAD65 was colocalized in a majority of the beta cells (> 95%), with only a few glucagon and somatostatin cells (< 5%) showing immunolocalization. During the fetal stages GAD65 also showed a similar predominant beta-cell coexpression. The enzyme was also detected in a few fetal glucagon (< 5%) but not somatostatin cells. In the degenerating large fetal islets, GAD65 was also observed in the majority of the residual beta cells. These results demonstrate that in the ovine pancreas GAD65 is expressed during fetal development and is predominantly beta cell-restricted. This pattern of expression is maintained during adult life. However, the physiological role of pancreatic GAD and/or its biosynthetic product, gamma-aminobutyric acid, in islet function in the sheep and in other ruminants remains unclear. PMID- 9204365 TI - Localization of a crustacean hyperglycemic hormone-like immunoreactivity in the neuroendocrine system of Euscorpius carpathicus (L.) (Scorpionida, Chactidae). AB - The neuroendocrine system of Euscorpius carpathicus was immunohistochemically localized using a polyclonal antiserum raised against a purified Homarus americanus crustacean hyperglycemic hormone (Hoa-cHHA). There were cross reactions in E. carpathicus procerebral and subesophageal neurosecretory cells, neurohemal organs, and intra- and extraganglionic neurosecretory tracts. Among the neurohemal structures, the Kwartirnikov's organ, the Tropfenkomplex, and the coxal disc reacted strongly. In Euscorpius, the differing results between adults and juveniles suggest neurosecretory variations related to developmental stage. These immunohistochemical observations suggest the presence of substances related to the cHH in scorpions; however, in this heterologous system, it is not at present possible to assess physiological significance. PMID- 9204366 TI - Arginine vasotocin gene expression and secretion during osmotic stimulation and hemorrhagic hypotension in hens. AB - In chickens, hyperosmolality stimulates the secretion of vasotocin (AVT) and up regulates hypothalamic AVT gene expression. Hemorrhage, on the other hand, has not been considered an effective stimulus for AVT release in this species. The effects of acute osmotic stress and prolonged hemorrhagic hypotension on AVT gene expression and secretion were studied in White Leghorn hens. Conscious hens were osmotically stimulated by administering a single ip injection of 3 M NaCl (5 ml/kg). Urethane-anesthetized hens were bled to a mean arterial pressure of 80-90 mm Hg and the pressure was maintained within this range by additional bleeding. A total of about 30% of the estimated blood volume was removed. Both experiments were terminated after 1 hr of stimulation. Plasma AVT levels in the hyperosmotic and hypovolemic hens were 4- and 2-fold higher, respectively, compared to controls. Hypothalamic AVT mRNA levels, detected by Northern blot analysis, were 2.5- and 2-fold higher in the osmotically stimulated and hypotensive groups, respectively, compared to control groups. As determined by in situ hybridization, both osmotic stimulation and hypovolemia resulted in an increase in the number of AVT mRNA-containing neurons in the supra-optic and paraventricular nuclei. Our results indicate that, under the conditions used, hypotension and hyperosmolality are equally effective in stimulating AVT gene expression and secretion of AVT. PMID- 9204364 TI - Neuroendocrine effects on adrenal hormone secretion in carp (Cyprinus carpio). AB - The responses of interrenal and chromaffin tissues of carp to acetylcholine (Ach) and its agonists/antagonists were studied in an in vitro perifusion system of head kidney. There was a dose-dependent release of epinephrine and norepinephrine to Ach between 0.01 and 100 mM added for 15 min to the incubation medium. Cortisol secretion was also stimulated, but the response peaked at 1.0 mM Ach and was attenuated with 10 or 100 mM Ach. The maximal release occurred about 30 min after addition of the transmitter. Nicotine stimulated the catecholamines, but had no effect on cortisol, while carbamylcholine, a nicotinic agonist, increased both the catecholamines and cortisol. Muscarine increased cortisol secretion, but affected catecholamines only at higher doses. In contrast, pilocarpine, a muscarinic agonist, stimulated catecholamines more than cortisol. Atropine was not antagonistic, rather it increased the secretion of catecholamines in a dose dependent manner, and inhibited the release of cortisol. It is concluded that both tissues are influenced by the autonomic nervous system, with the sympathetic system acting on chromaffin cells and the parasympathetic system acting on interrenal cells. However, the nerve supply cannot clearly be defined by agonists or antagonists as in mammals. There is evidence for paracrine effects, e.g., catecholamines inhibit cortisol release and cortisol influences catecholamine secretion. PMID- 9204367 TI - Distribution and characterization of natriuretic peptide receptors in the gills of the spiny dogfish, Squalus acanthias. AB - The distribution and nature of natriuretic peptide receptors (NPR) in the gills of dogfish, Squalus acanthias, were examined by tissue section autoradiography, competition analysis, protein electrophoresis, guanylate cyclase (GC) assays, and molecular cloning. Specific NP binding occurred on the gill filaments, but not on the interbranchial septum or gill arch. The binding was densest on the efferent edge of the gills. Higher resolution light-microscopic examination of emulsion coated sections showed that specific binding occurred mainly on the secondary lamellae and filament body and not on the arterial circulation. At least two types of NPR were revealed. One is linked to GC since NP binding stimulates the production of cGMP. The GC receptor may be similar to the NPR-B mammalian receptor since only pCNP stimulated cGMP production. The second receptor is not linked to GC and binds the specific ligand C-ANF [rat des(Gln18, Ser19, Gly20, Leu21, Gly22)]. The sequence of a cDNA generated using primers based on conserved regions of vertebrate NPR-C had considerable homology with mammalian and eel NPR C and eel NPR-D. The presence of GC-linked NPR and NPR-C/ NPR-D suggests that the gills are an important target organ for NP action. PMID- 9204368 TI - Interactions of androgens and estradiol on sex accessory ducts of larval tiger salamanders, Ambystoma tigrinum. AB - Immature tiger salamander larvae were treated with 12.5 or 25 micrograms of estradiol, testosterone, or dihydrotestosterone (DHT), or 12.5 micrograms of estradiol combined with 12.5 micrograms of either testosterone or DHT. Mullerian duct epithelium was more stimulated by combined steroid treatment than by any steroid alone. Estradiol antagonized the action of DHT in the Wolffian duct. Both of the androgens and estradiol when administered alone at the higher dose stimulated enlargement of connective tissue surrounding the ducts, but the combined 12.5 micrograms androgen/12.5 micrograms estrogen treatment was more effective even though the total steroid administered was the same. The effectiveness of DHT on mullerian cells of this species is evidence against a required aromatization of androgens to explain paradoxical steroid effects and suggests that fundamental differences may exist in steroid receptors of mullerian ducts, connective tissue, and Wolffian ducts. A possible role for the urodele duct system for assessing estrogenic activity of environmental contaminants is discussed. PMID- 9204369 TI - Ovarian steroid levels in Salamandra salamandra infraimmaculata during the reproductive cycle. AB - Gonadal steroid levels were determined in the ovary of Salamandra salamandra infraimmaculata during the reproductive cycle in populations from a xeric region in northern Israel. Varying proportions of previtellogenic and vitellogenic oocytes were present throughout the year, and mature oocytes were present in winter and spring. The numbers of mature oocytes were greater between December and April, after parturition. The levels of 17 beta-estradiol and testosterone rose during oocyte vitellogenesis and maturation. Levels of progesterone and 17 alpha-hydroxy progesterone appeared to be related to the level of vitellogenesis. Gravid females contained greater quantities of all four steroids than did nongravid females. PMID- 9204370 TI - The metabolism of biogenic monoamines during embryogenesis and metamorphosis in two anuran species. AB - This study investigated the pathways to many monoamines and their metabolites in the central nervous system of the frog Rana nigromaculata and the toad Bufo bufo japonicus during embryonic development and metamorphosis. Metabolites were analyzed by three-dimensional HPLC. The two species provided evidence of similar pathways, with slightly different timetables for the development of their monoamine systems. During embryonic development, the main metabolic pathways in entire embryos were tyrosine (TYR)-->[3,4-dihydroxyphenylalanine in Bufo]-->3 hydroxytyramine-->norepinephrine or epinine (EPIN)-->epinephrine, TYR-->tyramine- > (octopamine in Rana) and TYR-->3-O-methyldopa for catecholamines, and tryptophan-->kynurenine and 5-hydroxytryptamine (5-HT)-->[5-hydroxyindoleacetic acid and N-methyl-5-hydroxytryptamine (N-MET) in Bufo]. The monoamine system in the brain was similar during metamorphosis to that during embryogenesis with a few exceptions. The most striking change was the development of the bufotenine (5 hydroxy-N, N-dimethyltryptamine) pathway from 5-HT via N-MET. EPIN and norepinephrine in Rana and octopamine in both species disappeared during metamorphosis. These results are discussed in relation to the roles of the various pathways in development. PMID- 9204371 TI - Blastocyst development and conceptus sex selection in red deer Cervus elaphus: studies of a free-living population on the Isle of Rum. AB - Skewing of the sex ratio at birth occurs in red deer in response to dominance status, with dominant hinds giving birth to a higher proportion of male calves than subordinates. To investigate the physiological basis for this phenomenon, reproductive tracts were collected from red deer during a cull for management purposes carried out on the Island of Rum, Scotland. Blastocysts were flushed from the uterus and sexed by polymerase chain reaction using Y chromosome specific primers. Concentrations of interferon (measured as antiviral activity) in uterine flushings, of oxytocin receptors in endometrium, and of progesterone in jugular venous blood were measured, and ovarian morphology was recorded. Times of mating were determined retrospectively from calving dates observed during the following spring. Changes in uterine and fetal weights and sizes confirmed the degree of reproductive synchrony. Intervals between stages of blastocyst development (spherical, tubular, filamentous, and attached) derived from the observed incidence of each form showed that approximate times of blastocyst elongation and attachment were 13 and 30 days after conception, respectively. Hinds carrying male blastocysts were in better body condition (higher kidney fat weights, P = 0.025) than those carrying females. Interferon was detectable in uterine flushings from 1 of 7 hinds carrying early filamentous blastocysts and 5 of 12 hinds carrying late filamentous blastocysts, but in no case where the blastocysts were male (P = 0.035). Oxytocin receptor concentrations in caruncular endometrium (but not in intercaruncular endometrium) were lower in pregnant than in nonpregnant hinds (P < 0.05), but there was no correlation with interferon concentrations in flushings. Corpora luteal concentrations of oxytocin ranged from 1.8 to 51.2 micrograms/g tissue and declined with advancing blastocyst development. The data are consistent with the hypothesis that sexual dimorphism in trophoblast interferon production leads to differential blastocyst loss and hence to sex ratio skewing on the basis of dominance status. PMID- 9204372 TI - Functional comparison of mouse, rat, and fish islet grafts transplanted into diabetic nude mice. AB - Equal volumes of teleost fish (tilapia), Lewis rat, or CD-1 mouse islets were transplanted under the kidney capsules of streptozotocin-diabetic athymic nude mice. Nonfasting blood glucose levels were monitored in recipient mice over a period of 30 days. Mean nonfasting blood glucose levels in recipients of tilapia (n = 7), rat (n = 8), and murine (n = 8) islets were 78.8, 77.0, and 115 mg/dl, respectively. Mean blood glucose levels were significantly higher in recipients of murine islets than in recipients of fish and rat islets. After Day 30, intraperitoneal glucose tolerance tests were performed on recipient mice. Mean fasted blood glucose levels in mouse, rat, and fish islet recipients were 113.3, 89.8, and 72.7 mg/dl, respectively. All three groups of recipient mice had similar glucose tolerance profiles with mean glucose disappearance rates (K values) between 4.3 and 5.7. Tilapia islet grafts resulted in a significantly lower baseline for blood glucose values than either rat or mouse islet grafts. PMID- 9204373 TI - Isolation and characterization of two distinct gonadotropins from the pituitary gland of Mediterranean yellowtail, Seriola dumerilii (Risso, 1810). AB - Two gonadotropins, GTH I and GTH II, were isolated and chemically characterized from the pituitary of Mediterranean yellowtail. They were extracted with 35% ethanol-10% ammonium acetate, separated by ion-exchange chromatography on a DE-52 column, and purified by reversed-phase high-performance liquid chromatography on Asahipak C4P-50 and subsequently by gel filtration chromatography on Superdex 75. The molecular weights were estimated at 47 kDa for GTH I and 29 kDa for GTH II by SDS-PAGE and at 49 kDa for GTH I and 42 kDa for GTH II by gel filtration. GTH II was completely dissociated, while GTH I was partially dissociated into alpha- and beta-subunits by treatment with 0.1% trifluoroacetic acid. The complete amino acid sequences of GTH alpha-, GTH I beta-, and GTH II beta-subunits were determined. The GTH alpha-subunit consisted of 91 amino acid residues. The GTH I beta and GTH II beta consisted of 105 and 115 amino acid residues, respectively, and had a 28% sequence identity to each other. They had the highest sequence identity with the respective gonadotropin subunits of bonito, tuna, and striped bass: 81-83% for GTH alpha, 67-71% for GTH I beta, and 91-93% for GTH II beta. The sequence identity of the GTH alpha-subunit with those of other teleosts and human and bovine LH and FSH was 57-67%. The GTH I beta-subunit showed a low sequence identity with other known fish GTH I beta s (36-51%) and was more similar to human and bovine FSH beta s (34% identity) than to human and bovine LH beta s (29% identity). The sequence identity of the GTH II beta-subunit with those of other teleosts was higher (60-73%), being more similar to LH beta s (43% identity) than FSH beta s (38% identity). Thus, two distinct gonadotropins, GTH I and GTH II, homologous to mammalian FSH and LH, respectively, are synthetized by M. yellowtail pituitary glands. PMID- 9204374 TI - Subcellular localization of 3 beta hydroxysteroid dehydrogenase isomerase in testis of Bufo arenarum H. AB - 3 Beta-hydroxysteroid dehydrogenase 5-ene isomerase (3 beta HSD/I) catalyzes an essential step in the biosynthesis of steroid hormones and is usually considered to be mainly microsomal, although there is a dual distribution of the enzyme in toad interrenals. The present study demonstrates that in the testicular tissue, as in interrenals of Bufo arenarum H., 3 beta HSD/I is both mitochondrial and microsomal. The conversion of dehydroepiandrosterone to androstenedione takes place only in microsomes while pregnenolone is converted to progesterone in both microsomes and mitochondria. Kinetic constants of 3 beta HSD/I were determined by the oxidation of pregnenolone and dehydroepiandrosterone. The preferred substrate of the microsomal 3 beta HSD/I enzyme was dehydroepiandrosterone (K(m) = 0.17 microM and 0.53 microM for dehydroepiandrosterone and pregnenolone, respectively) not only during the breeding season but also in the non-breeding period (K(m) = 0.49 microM and 2.9 microM for dehydroepiandrosterone and pregnenolone, respectively). PMID- 9204375 TI - Estradiol inhibits plasma somatostatin 14 (SRIF-14) levels and inhibits the response of somatotrophic cells to SRIF-14 challenge in vitro in rainbow trout, Oncorhynchus mykiss. AB - In the present study, the effects of 17 beta-estradiol (E2) treatment on plasma growth hormone (GH) and somatostatin 14 (SRIF-14) concentrations were investigated, as well as the effect of in vivo E2 treatment on the in vitro GH response to SRIF-14 challenge in sexually immature rainbow trout (Oncorhynchus mykiss). Two weeks after receiving a steroid hormone implant, plasma E2 and GH levels were significantly (P < 0.05) elevated, and plasma SRIF levels were significantly (P < 0.05) lowered relative to the control. Pituitary glands were taken from E2-primed and control fish and challenged with a single pulse of SRIF 14 (10(-8) M) in a perifusion unit to evaluate the effect of E2 on the response of somatotrophs to the effect of SRIF-14. Whereas SRIF-14 challenge significantly (P < 0.01) inhibited GH release from pituitary fragments taken from control fish, there was no such response in E2-primed fish. Furthermore, GH release following SRIF-14 administration (at the point of maximal inhibition) was significantly depressed in control fish with respect to the E2 treatment group. These data suggest that E2 treatment may increase plasma GH concentrations by altered somatotroph responsiveness to SRIF-14 inhibition. Furthermore, E2 may increase plasma GH by suppressing plasma SRIF-14 levels, although the role of circulating SRIF-14 on the regulation of GH release has not been fully determined in teleosts. PMID- 9204376 TI - Melatonin levels in the gastrointestinal tissues of fish, amphibians, and a reptile. AB - Melatonin was detected by radioimmunoassay in the gastrointestinal tract (GIT) of several species of fish (sturgeon, rainbow trout, carp), amphibians (axolotl, leopard frog, bullfrog), and one reptile (red-sided garter snake), which were sacrificed during the daytime. The highest levels of melatonin were detected in the snake [means = 1018 pg/g stomach, 328 pg/g proximal gut (PG), 511 distal gut (DG)] and carp (means = 102 pg/g stomach, 146 pg/g PG and 141 pg/g DG). Lowest levels were found in the axolotl (means = 44 pg/g stomach and PG, 92 pg/g DG) and the bullfrog (means = 73 pg/g esophagus, 78 pg/g stomach, 20 pg/g PG, and 152 pg/g DG). In most cases there were no statistically significant differences in the melatonin levels among various GIT tissues of the same species but there were differences in tissue levels between different species. PMID- 9204377 TI - Optimum sarcomere length in mammalian diaphragm. PMID- 9204378 TI - Quantitative structure-activity relationship studies on some anti-human immunodeficiency-virus-1 (anti-HIV-1) drugs: viral reverse transcriptase inhibitors. AB - The anti-HIV-1 activity of some 3-[(benzoxazol-2-ylmethyl)amino]-, 3-[(benzoxazol 2-yl)ethyl]-, 3-[N-(phthalimidomethyl)amino]- and 3-[N-(phthalimido)ethyl]-5 ethyl-6-methyl pyridin-2(1H)-one derivatives, that have been found to elicit their action through the allosteric inhibition of the enzyme viral reverse transcriptase (VRT), have been analysed in relation to the physicochemical properties of the molecules. Significant correlations were obtained between the activity and the hydrophobic and electronic constants of substituents and van der Waals' volume of the linker chain. Based on these findings the mechanism of action of these drugs is discussed. PMID- 9204379 TI - Cholinesterase inhibition by derivatives of 2-amino-4,6-dimethylpyridine. AB - Derivatives of 2-amino-4,6-dimethylpyridine, aryl(alkyl)carboxamides, thiocarbamides and amidrazones, already known for their anti-inflammatory properties, were found to be moderately active inhibitors of acetyl and butyrylcholinesterase. Quantitative structure-activity relationships showed that the binding affinity was enhanced by the following structural modifications: (1) increase in molecular volume, (2) decrease in the energy of the lowest unoccupied molecular orbital, (3) insertion of a methylene group between the amide carbonyl and the aromatic ring, (4) replacement of the amide oxygen by sulfur. The affinity remained, however, weaker than that of the specific inhibitor 9-amino 1,2,3,4-tetrahydroacridine (tacrine). The association of anti-inflammatory and cholinesterase inhibiting activities within the same compound may prove useful for the treatment of Alzheimer's disease. PMID- 9204380 TI - Inhibition of octapeptide N-myristoylation by acyl amino acids and acyl alkanolamines. AB - Several acyl amino acids and acyl alkanolamines were prepared and screened for their inhibition of octapeptide N-myristoylation and HIV-1 replication in MT-4 cells. Of the 62 acyl derivatives tested, N-myristoyl-O-caproyl-L-serine, N myristoyl-O-caproyl-D-serine and N-decanoyl-O-myristoyl-L-serine were found to be uncompetitive inhibitors of N-myristoylation, but did not prevent HIV-induced cytopathicity in MT-4 cells. However, other acyl derivatives such as N-3 hydroxymyristoyl ethanolamine, N-3-hydroxymyristoyl-D-serine and N-myristoyl-L cysteine, which did not inhibit N-myristoylation, suppressed the cytopathicity in the infected cells. The acyl derivatives described here may serve as lead compounds for antiviral agents. PMID- 9204381 TI - Novel aromatic/heterocyclic sulfonamides and their metal complexes as inhibitors of carbonic anhydrase isozymes I, II and IV. AB - Reaction of five aromatic/heterocyclic sulfonamides containing free amino, groups with 5-nitro-alpha-2-toluenesultone and diethyl pyrocarbonate, respectively, afforded novel inhibitors of the zinc enzyme carbonic anhydrase (CA). Zn(II) complexes of the new sulfonamides were prepared. Excellent inhibition of three CA isozymes (CA I, II and IV respectively) were observed with some of the new sulfonamides, but especially with their Zn(II) complexes. Structure-activity correlations in this series of inhibitors are discussed. PMID- 9204382 TI - Differentiation between various types of inotropes through discovery of differences in their ability to detect isoforms of Na+/K(+)-ATPase. PMID- 9204383 TI - The mechanism of a P-450 enzyme-aromatase; a molecular modelling perspective for the removal of the C(19) methyl and aromatisation of the steroid A ring. AB - The three possible mechanisms for the final step of aromatisation, as proposed by Wright and Akhtar, are studied using a molecular modelling approach utilising the 'substrate-heme complex' previously reported. The study considers the three mechanisms from a geometric point of view and concludes that only a ferroxy radical is involved in all three steps of aromatase action and not the mixture of both ferroxy and peroxy as previously suggested. The study also suggests that an alternative peroxy-based mechanism is unlikely due to the distances between reacting species. An alternative theoretical mechanism, which circumvents the production of the CHO. radical (regarded to be too small to be retained within the active site) for the final step of aromatisation is suggested involving the concerted breakup of the iron-formate complex together with hydrogen abstraction from C(1) of 3-hydroxy-estra-2,4-diene-17-one, resulting in oestrone and formic acid. PMID- 9204384 TI - Phosphonamide inhibitors of neutral endopeptidase (EC 3.4.24.11). PMID- 9204385 TI - Inhibition of dipeptidyl peptidase IV (CD26) by peptide boronic acid dipeptides. AB - Peptide boronic acid dipeptide compounds were analyzed for their ability to inhibit recombinant human dipeptidylpeptidase IV (CD26, DPPIV). Rate constants for the peptide boronates are difficult to obtain because the active boronic acid dipeptide exists in equilibrium with a cyclic inactive species in aqueous solution. Rate constants were determined for the inhibition of DPPIV using several peptide boronates at different pH values. Val-boroPro forms the most tightly bound complex with DPPIV; the first order half life for dissociation of the inactive enzyme-inhibitor complex at 23 degrees C is approximately 27 days. PMID- 9204386 TI - Quantitative structure-activity relationship studies on some viral reverse transcriptase inhibitors acting as anti-HIV-1 agents. AB - The anti-HIV-1 and cytotoxic activities of some viral reverse transcriptase inhibitors, namely the analogues of [1-[2',5'-bis-O-(tert-butyldimethylsilyl) beta-D-xylo- and -ribofuranosyl]]-3'-spiro-5" -[4"-amino-1",2"-oxathiole 2",2" dioxide] (TSAO) pyrimidine and pyrimidine modified-nucleotides, are analysed in relation to their physicochemical and molecular properties. The antiviral activities of the compounds are found to be significantly correlated with hydrophobic and electronic properties of the molecules, but no physicochemical parameters were found to be correlated with the cytotoxic effects of the compounds. This difference is exploited to improve the selectivity of the compounds. It is observed that TSAO can provide potent anti-HIV-1 drugs with a disubstituted thymine ring, in which a substituent may be at the N3-position. The disubstitution reduces the cytotoxicity, and substituents' hydrophobicity and electron donating character enhance the antiviral activity. PMID- 9204387 TI - Synthetic macromolecular inhibitors of human leukocyte elastase. 2. Effect of loading of a peptidyl carbamate inhibitor and molecular size of polymer backbone on its inhibitory capacity. AB - Several macromolecular inhibitors of human leukocyte elastase (HLE) were prepared by covalently bonding a low molecular weight HLE inhibitor peptidyl carbamate, p nitrophenyl-N-[succinyl-L-alanyl-L-ananyl-L-prolylmethyl]- N-isopropyl carbamate 1, with the neutral hydrophilic polymer, poly-alpha,beta-[N-(2-hydroxyethyl)-D,L aspartamide], PHEA 2. These novel polymeric compounds differed in the molecular size of their PHEA polymer backbone and the extent of loading of the peptidyl carbamate, (PC). They were shown to efficiently inhibit HLE (Ki = 97 to 12.8 nM) as intact macromolecular entities and were found to be more stable to hydrolysis than the non-polymer bound low molecular weight inhibitor 1. The inhibition of HLE by the novel macromolecular inhibitors was found to be noncompetitive and reversible, proceeding via slow formation of inhibitor-enzyme complex. The effect of loading of 1 and molecular size of the PHEA 2 polymer on enzymatic parameters Ki, kon and koff is discussed and a possible mechanism of inhibition is presented. PMID- 9204388 TI - Mechanism of eserine action on the hydrolysis of butyrylthiocholine by butyrylcholinesterase. AB - The mechanism of the interaction of eserine with butyrylcholinesterase has been proposed only on the basis of analogy with acetylcholinesterase. Here the interactions was studied in detail and the results analysed by classical kinetic methods and by means of mathematical modelling. An appropriate kinetic scheme was developed, an adequate equation derived and the corresponding kinetic parameters evaluated. The findings suggest that a fast but relatively weak binding of eserine to the enzyme's active site is followed by a slow acylation step and by an even slower rate limiting deacylation step so misrepresenting eserine as an irreversible inhibitor. The proposed kinetic scheme also suggests that the reaction of eserine with a peripheral substrate site is unlikely as seen with the substrate, butyrylthiocholine. PMID- 9204389 TI - Evidence for the essential histidine at the NADPH binding site of enoyl-CoA reductase domain of pigeon liver fatty acid synthetase. AB - Pigeon liver fatty acid synthetase was inactivated by stoichiometric concentrations of diethylpyrocarbonate (DEP). The second order rate constant for the loss of synthetase activity was similar to the value for enoyl-CoA reductase indicating that ethoxyformylation destroys the ability of the enzyme to reduce the unsaturated acyl intermediate, without significant effect on beta-ketoacyl reductase activity. NADPH provided protection to the enzyme against inactivation by DEP indicating that essential histidine residues are present at the active site. DEP-modified enzyme showed a characteristic absorption maxima at 240 nm confirming the formation of ethoxyformic histidine. The reversal of inactivation by hydroxylamine and a pKa value of 7.0 obtained from the pH-rate profile for inactivation again confirmed the specificity of DEP for histidine. Stoichiometric results showed that two moles of histidine residue per mole of enzyme are essential for the activity of FAS. PMID- 9204390 TI - A simple novel method for studying the combined inhibitory effects of ethylurea and N,N-dimethylurea on jack bean urease. AB - The kinetics of the inhibition of jack bean urease in the presence of ethylurea and N,N-dimethylurea was studied at pH = 7.0. Both inhibitors were competitive inhibitors. A simple novel method was used for determining the dissociation constants (Ki) values for the inhibition which were 26 and 28 mM respectively. PMID- 9204391 TI - Inhibitory effect of selenium on enzymes involved in heme biosynthetic pathway in chick embryos. AB - Effect of different concentrations of selenium (Se) on heme biosynthesis was studied at different developmental stages of chick embryo. The first rate limiting enzyme ALA-synthase (ALA-S; E.C.2.3-1.37) activity was enhanced by selenium, while hepatic and blood ALA-dehydratase activity (ALA-d; E.C.3.2.1.24) was decreased. Hepatic and blood free-sulfhydryl (-SH) group contents were significantly decreased by Se. Further, hepatic aminolevulinic acid (ALA) and total blood porphyrin levels were enhanced and hepatic heme levels were depleted by selenium exposure. Heme biosynthesis was maximally inhibited in the E4 (4th day injected embryos) when compared to later periods. PMID- 9204392 TI - Inhibition kinetics of camel lens zeta-crystallin: multiple inhibition studies. AB - The inhibition of camel lens zeta-crystallin by nitrofurantoin (NF) was uncompetitive with respect to co-factor NADPH, (Ki = 90 microM) and competitive with respect to the substrate 9,10-phenanthrenequinone (PQ), (Ki = 50 microM). Inhibition at micromolar concentrations was also observed with dicoumarol, NADP+ and cibacron blue (CB). Theorell-Yonetani double-inhibition analysis showed that NF and dicoumarol were mutually exclusive inhibitors against PQ. However, analysis of NF and NADP+ by a double-inhibition plot showed that they simultaneously bind to the enzyme molecule. These studies demonstrate that NF and dicoumarol share the same site so that both molecules are prevented from binding at the same time, while NF and NADP+ can bind simultaneously to different sites on the enzyme. Although CB was noncompetitive with respect to PQ, double inhibition analysis showed that CB and dicoumarol or NF were mutually exclusive inhibitors against PQ, implying a distinct mode of inhibition for CB. PMID- 9204393 TI - Quantitative structure-activity relationship studies on anti-HIV-1 TIBO derivatives as inhibitors of viral reverse transcriptase. AB - The anti-human-immunodeficiency-virus (HIV-1) activity of the derivatives of 4,5,6,7-tetrahydro-5-methylimidazo [4,5,1-jk] [1,4] benzodiazepin-2(1H)-one (TIBO) that have been found to elicit their action through the allosteric inhibition of the enzyme viral reverse transcriptase (VRT) is analysed in relation to the physicochemical properties of the molecules. Significant correlations are obtained between the activity and the hydrophobic constant and some dummy parameters of substituents. Based on these findings, the mechanism of action of these anti-HIV drugs is discussed. PMID- 9204394 TI - Effect of YM-47141, a new inhibitor produced by Flexibactor sp. Q17897, on elastase. AB - YM-47141, a peptidic compound recently isolated from Flexibactor sp. Q17897, strongly inhibited human leukocyte elastase (HLE) with Ki value 2.1 x 10(-7) M. Unlike other serine protease inhibitors, YM-47141 exhibited relatively weak effects on cathepsin G and alpha-chymotrypsin and its inhibitory Ki values were 9.2 x 10(-4) M and 1.3 x 10(-6) M, respectively. It had little, or no inhibitory effect on plasmin, thrombin, trypsin and kallikrein (IC50 > 10(-4) M). The inhibition of HLE by YM-47141 was reversible and of a mixed type. PMID- 9204395 TI - A novel synthetic inhibitor of endopeptidase-24.15. AB - A novel synthetic inhibitor of endopeptidase-24.15 (EP-24.15, EC 3.4.24.15), N [(2R,4R)-2-(2-hydroxyphenyl)-3-(3-mercaptopropionyl)-4-thiazolidine carbonyl] -L phenylalanine (SA898) is described. This compound inhibited rat EP-24.15 competitively with an IC50 of 23 nM and Ki of 9.1 nM. These values were, respectively, 9.6 times and 6.3 times smaller than those for N-(1-carboxy-3 phenylpropyl)-alanyl-alanyl-phenylalanyl-p-aminobenzoate (cFP-AAF-pAB), which was one of the most potent inhibitors thus far reported. The inhibitory effect of SA898 on other endopeptidases, angiotensin converting enzyme (ACE, EC 3.4.15.1) and endopeptidase-24.11 (EP-24.11, EC 3.4.24.11) was also studied. SA898 inhibited ACE significantly, but the potency was about 20-fold lower than that for EP-24.15 in terms of the Ki value. The inhibitory effect of SA898 on EP-24.11 was almost negligible (Ki = 28 microM). In addition, the inhibitory activities of several SA898-related compounds were examined. Based on these data, the structure activity relationships for EP-24.15 inhibitors are discussed. PMID- 9204396 TI - Piperastatin B: a new selective serine carboxypeptidase inhibitor from Streptomyces lavendofoliae MJ908-WF13. AB - Piperastatin B, a new inhibitor of serine carboxypeptidase was purified from a culture broth of Streptomyces lavendofoliae MJ908-WF13 as a minor component by monitoring its inhibitory activity against carboxypeptidase Y (CP-Y). Its structure was determined to be N-formyl-Val-Thr-Leu-Val-Pip-Leu-Pip (pip: piperazic acid, hexahydropyridadine-3-carboxylic acid). Piperastatin B is a highly specific competitive inhibitor of CP-Y (Ki = 55 nM) with little effect on related enzymes and resembles the major component, piperastatin A, in these respects. PMID- 9204397 TI - Alkylphosphate esters as inhibitors of phospholipase D. AB - Alkylphosphate esters were shown to be potent inhibitors of phospholipase D. Using phosphatidyl choline/sodium dodecylsulfate (2:1) as substrate, IC50 values were determined for alkylphosphocholines of different chain length (C10-C18) and for various octadecylphosphate esters with different polar head groups. The inhibitory potency strongly increased with increasing chain length of the alkyl chain. The substitution of choline for heterocyclic nitrogen compounds or for 2 trimethylarsonio-ethanol also affected the inhibition of phospholipase D. Octadecylphosphocholine proved to be the most efficient inhibitor (IC50 = 6.4 microM). PMID- 9204398 TI - Inhibitors of the Ras signal transduction pathway as potential antitumour agents. PMID- 9204399 TI - Inhibiting effects of spermidine derivatives on Trypanosoma cruzi trypanothione reductase. AB - Trypanothione reductase is a vital component of the antioxidant defenses of trypanosomes. This enzyme reduces trypanothione, a spermidine-glutathione conjugate. The inhibitory effects of several spermidine derivatives on the reduction of trypanothione by Trypanosoma cruzi trypanothione reductase were assessed. Spermidine derivatives containing hydrophobic aromatic substituents were found to be competitive inhibitors of trypanothione reductase. N4-acylated spermidine derivatives were less effective inhibitors than the corresponding N4 alkylated derivatives. The most effective compounds studied were N1, N8-bis(2 naphthylmethyl)spermidine and N4-(2-naphthylmethyl)spermidine, with Ki values of 9.5 and 108 microM, respectively. PMID- 9204400 TI - Effects of YM-51084 and YM-51085, new inhibitors produced by Streptomyces sp. Q21705, on cathepsin L. AB - The structures of YM-51084 and YM-51085, new protease inhibitors produced by Streptomyces sp. Q21705, were determined by 1H- and 13C-NMR and mass spectrometry. Both were characterized by the basic structures of an acyl tripeptide. YM-51084 was elucidated to be isovaleryl-tyrosyl-valyl-phenylalaninal and YM-51085 was the reduced phenylalaninol form of YM-51084. These compounds proved to strongly inhibit human kidney cathepsin L; the IC50 values being 9.6 x 10(-9) M and 3.5 x 10(-7) M, respectively. PMID- 9204401 TI - Kinetics of the inhibition of acetylcholinesterase from desert cobra (Walterinnesia aegyptia) venom by local anesthetics: procaine and tetracaine. AB - The kinetic parameters of W. aegyptia venom acetylcholinesterase (AChE) inhibition by procaine and tetracaine hydrochloride were investigated in the present study. Procaine and tetracaine reversibly inhibited the AChE activity in a concentration-dependent manner, the IC50 being about 0.28 and 0.04 mM, respectively. The Michaelis-Menten constant (K(m)) for the hydrolysis of acetylthiocholine iodide was found to be 0.051 mM with Vmax 10.2 mumole/min/mg protein. Both K(m) and Vmax were affected by procaine while only Vmax decreased with tetracaine. A Lineweaver-Burk plot and its secondary replot indicated that the nature of the inhibition is of the linear mixed type for procaine which is considered to be a mixture of competitive and noncompetitive types while the inhibition was noncompetitive for tetracaine. The values of Ki(slope) and K(intercept were estimated as 0.133 mM and 0.451 mM for procaine and 7.2 x 10(-3) mM for tetracaine, respectively, by the secondary replots of the Lineweaver-Burk plot. PMID- 9204403 TI - Inactivation of yeast glutathione reductase by O-phthalaldehyde. AB - Yeast glutathione reductase was inactivated by the bifunctional reagent, o phthalaldehyde. The initial rate of inactivation followed pseudo-first order kinetics. Fluorescence spectral properties of modified enzyme indicated the formation of an isoindole derivative from cysteine and lyaine residues present in close proximity as shown by typical fluorescence emission and excitation maximum at 410 nm and 337 nm, respectively. The fluorescence spectral studies with o phthalaldehyde in the presence and absence of N-ethylmaleimide indicated that both the inhibitors react with the same cysteine residue, which is non-essential for enzyme activity. The coenzyme NADPH did not protect the enzyme against the o phthalaldehyde reaction while oxidised glutathione prevented o-phthalaldehyde inactivation. This could be due to reaction of the amino group of glutathione with o-phthalaldehyde. Stoichiometry of the reaction showed that the formation of approximately 2 isoindole derivatives per subunit of glutathione reductase is accompanied by 75% loss of activity. The results suggest that o-phthalaldehyde binds to non-essential cysteine and lysine residues present in close proximity which results in conformational changes leading to enzyme inactivation. PMID- 9204402 TI - Inhibitory action of the defensive discharge of the grasshopper, Poecilocerus pictus, on certain enzymes in the lizard, Calotes nemoricola. AB - Administration of the defensive secretion of the grasshopper, Poecilocerus pictus inhibited acetyl-cholinesterase (AChE) and adenosine triphosphatases (ATPases) in the brain and muscle tissues of the garden lizard, Calotes nemoricola. The inhibition was gradual, continuous and irreversible with lethal doses of the defensive secretion, whereas the inhibition observed with sublethal doses was followed by an increase towards control levels within 24 h after injection. In vitro application of defensive secretion also showed concentration-dependent inhibition in the activity of AChE and ATPases in the tissue homogenates. Inhibition in AChE activity might be a factor for the observed mortality is the defensive fluid-treated lizards. Since the cardenolides are known to inhibit the activity of ATPases, the inhibition in the activity of ATPases observed in the present study suggests the presence of cardenolides in the defensive fluid of P. pictus. PMID- 9204404 TI - Liver apoptosis. AB - Apoptosis, also called programmed cell death, is a peculiar form of cell death different from cell necrosis in many morphological and biochemical aspects. Like mitosis or differentiation, apoptosis is a normal cell phenomenon which depends on the expression of genes capable of inducing or inhibiting this type of cell destruction. But apoptosis can also be triggered by many external factors and has been described in many diseases. The very different conditions where programmed cell death occurs suggest that the mechanisms leading to the activation of apoptosis-controlling genes are variable. As in other cells, apoptosis occurs in the liver cells, first in the normal state during liver development and then in the adult liver, respectively for liver organogenesis and the renewal of hepatocytes. But apoptosis is also present in various viral, immunological, malignant or drug-induced human liver diseases. In addition, in the animal, hepatocyte apoptosis can be triggered either in vivo or in vitro by many toxic agents. In contrast to other cells, the mechanisms leading to liver cell apoptosis remain poorly investigated. However, two proteins could play an important role in this field, the fas/apo-1 protein present at the surface of hepatocytes and the bcl-2 protein localized in biliary cells. Analysis of the genes controlling the expression of these two proteins could provide essential information on the mechanisms of liver apoptosis. PMID- 9204405 TI - Genetic predisposition to drug-induced hepatotoxicity. AB - Drug-induced hepatitis is uncommon and generally unpredictable. Hepatotoxicity may be related to the drug itself, or to chemically reactive metabolites which can bind covalently to hepatic macromolecules and may lead to either idiosyncratic, toxic hepatitis or to immunoallergic hepatitis. There is now evidence indicating that genetic variations in systems of biotransformation or detoxication may modulate either the toxic or sensitizing effects of some drugs. Thus, the genetic deficiency in a particular hepatic cytochrome P 450 isozyme (CYP 2D6) is involved in per-hexiline liver injury. The deficiency in CYP 2C19 might also contribute to Atrium hepatotoxicity. Slow acetylation related to N acetyltransferase 2 deficiency contributes to sulfonamide hepatitis. The genetic deficiency in glutathione synthetase may increase the susceptibility to several drugs including acetaminophen. A constitutional deficiency in another cell defense mechanism, still not characterized, seems to increase significantly the risk of hepatotoxicity with halothane, phenytoin, carbamazepine, phenobarbital, sulfamides and amineptine. PMID- 9204406 TI - Molecular structure and hepatotoxicity: compared data about two closely related thiophene compounds. AB - Two closely related compounds, a diuretic drug tienilic acid (TA) and its isomer TAI have been found to exert very different toxic effects. In human liver microsomes TA is oxidized mainly by CYP 2C9 with formation of a reactive metabolite which covalently binds to CYP 2C9 in a rather specific manner. On the contrary, CYP 2C9-dependent oxidation of TAI leads to reactive metabolite(s) causing an intense covalent binding to several microsomal proteins. Based on these very different behaviours and fates of TA and TAI metabolites, it is proposed that the direct hepatotoxic effects of TAI could be due to an intense, non-specific covalent binding of its reactive metabolite(s) to liver proteins, whereas the toxic effects of the immunoallergic type of TA could be due to the very specific covalent binding of its sulfoxide metabolite to CYP 2C9. PMID- 9204407 TI - Hepatotoxicity in drug development: detection, significance and solutions. AB - Despite considerable progress in the understanding of the mechanism of liver toxicity we are not yet able to design non-hepatotoxic molecules rationally. Also, there is no "universal" in vitro primary screening approach for early identification of hepatotoxic molecules. In most cases hepatotoxicity is detected at later stages of drug development in animal toxicity studies or clinical trials. Although the liver is the most common target organ for drug candidates in animal toxicity studies, hepatotoxicity rarely leads to cessation of drug development during the preclinical phase. Indeed, contrary to other target organs, liver toxicity is usually reversible and can be monitored in man by sensitive serum enzyme tests. Therefore in many cases a compound found hepatotoxic in an animal species will be tested in man for a definitive assessment of its hepatotoxic potential. Liver toxicity in man may be acceptable when a drug has major therapeutic potential. In this situation mechanistic studies are essential to assess the risk in man and in some cases to identify protective agents. When liver toxicity leads to project termination a secondary screening approach may be envisaged if biologically active analogs are available. PMID- 9204408 TI - Immunotoxicology of the liver: adverse reactions to drugs. AB - Liver is a frequent target for drug-induced hepatitis. They can be classified in two categories: the hepatitis in which the drug or a metabolite reach a vital target in the cell and the hepatitis in which the drug triggers an adverse immune response directed against the liver. We will discuss essentially this second kind of disease. They have key clinical features such as the low frequency, the dose independence, the delay between the beginning of drug intake and the triggering of the disease, the shortening of the delay upon rechallenge and very often the presence of autoantibodies in the serum of the patients. Such signs were found in hepatitis triggered by drugs such as halothane, tienilic acid, dihydralazine, anticonvulsants. They will be taken as examples to show the recent progress in the understanding of the mechanisms leading to the disease. It has been postulated that the drug is metabolised into a reactive metabolite binding to the enzyme which generated it; therefore the neoantigen might trigger an immune response characterised by the production of antibodies recognising the native and or the modified protein. Most of these steps were proven in the cases of halothane, tienilic acid and dihydralazine. Several points seem important in the development of the disease; the equilibrium between toxication and detoxication pathways, the nature and amount of neoantigen, the individual immune response. However, many points remain unclear: for instance, the reason for the very low frequency of this kind of disease; the precise mechanism of the adverse immune response; the risk factors for developing such adverse reactions. Efforts should be made to better understand the mechanisms of this kind of disease: for instance, an animal model, tests to identify drugs at risk for such reactions, the role of these drugs in the processing of P450s and the processing of the neoantigens for their presentation to the immune system. PMID- 9204409 TI - Impaired mitochondrial function in microvesicular steatosis. Effects of drugs, ethanol, hormones and cytokines. AB - Microvesicular steatosis occurs in conditions characterized by severe impairment of the mitochondrial beta-oxidation process, due to genetic and/or acquired causes. Drugs and some endogenous compounds can sequester coenzyme A (aspirin, valproic acid), inhibit mitochondrial beta-oxidation enzymes (tetracyclines, several 2-arylpropionate anti-inflammatory drugs, amineptine and tianeptine), or inhibit both mitochondrial beta-oxidation and oxidative phosphorylation (endogenous bile acids, amiodarone, perhexiline and diethylaminoethoxyhexestrol), while female sex hormones have complex, but moderate, effects on mitochondrial structure and function. Other substances impair mitochondrial DNA transcription (interferon-alpha) or mitochondrial DNA replication (dideoxynucleosides), while alcohol abuse might accelerate the normal oxidative aging of mitochondrial DNA. When beta-oxidation is severely impaired, fatty acids, which are poorly oxidized by mitochondria, are mainly esterified into triglycerides, but there is a residual increase in non-esterified fatty acids. Triglycerides (possibly emulsified by a rim of non-esterified fatty acids) accumulate as small vesicles. Impairment of energy production, and the mitochondrial and general toxicity of both non-esterified fatty acids and dicarboxylic acids, may contribute to liver failure, coma and death in severe forms. Although milder forms of microvesicular steatosis have a good short-term prognosis, they can lead to chronic lipid peroxidation and the development of steatohepatitis lesions. Investigational molecules with a carboxylic group or a protonatable amine, or those which might interfere with mitochondrial DNA, should be screened for possible mitochondrial effects. PMID- 9204410 TI - Role of the liver-enriched transcription factors C/EBP alpha and DBP in the expression of human CYP3A4 and CYP3A7. AB - In the human fetal liver, CYP3A7 is expressed as early as the 13th week of gestation. This continues to the perinatal period when it is sharply repressed prior to birth. Concomitantly, the expression of CYP3A4, not detectable in the fetus, sharply increases in the perinatal period to remain elevated throughout adulthood. The mechanisms controlling these developmental patterns of expression have not yet been elucidated at the molecular level. The aim of the present work was to make a functional analysis of the 5'-flanking regions of CYP3A4 and CYP3A7 in different cell lines, including CHO, HepG2, WRL68 and Caco-2 TC7, after cotransfection with two hepato-specific transcription factors, C/EBP alpha and DBP. Six deletions of different length of the 5'-flanking region of each gene, spanning from -1240 to +11 for CYP3A4 and from -1157 to +13 for CYP3A7, were analysed by reporter gene assay. With the CYP3A4 constructs, C/EBP alpha stimulated the transcriptional activity in CHO cells in a way that suggested the presence of at least two C/EBP alpha-responsive elements, one downstream of -55 and one upstream of this position. In CYP3A7, the proximal element exhibited comparable stimulation to the corresponding one in CYP3A4, although the more distal one appeared to respond to a much smaller extent. CYP3A4 and CYP3A7 constructs also responded to C/EBP alpha in HepG2 and WRL68. However, only CYP3A4 and not CYP3A7 was transactivated by this factor in the Caco-2-TC7 cell line. In CHO cells, only the shortest proximal promoter deletion of CYP3A4 (downstream of 57) responded to DBP, while neither the longer constructs nor the CYP3A7 deletions were transactivated. Although preliminary, our results suggest that C/EBP alpha, and possibly other members of the C/EBP family, play a prominent part in the expression of the CYP3A family in man, and that the two genes respond differently to C/EBP alpha and DBP, two factors that exhibit a strict proliferation-dependent pattern of expression in the liver. PMID- 9204411 TI - Drug elimination in chronic liver diseases. PMID- 9204412 TI - Use of hepatocyte cultures for the study of hepatotoxic compounds. AB - Human and animal hepatocytes in primary culture are widely used in pharmacotoxicological research. They represent a unique in vitro model since they retain both phase I and phase II enzyme activities as well as their inducibility by xenobiotics. Hepatocyte cultures are used for drug screening, identification of the lesions induced by toxic compounds and determination of mechanisms by which xenobiotics exert liver injury. PMID- 9204413 TI - Simulation of human xenobiotic metabolism in microorganisms. Yeast a good compromise between E. coli and human cells. AB - An overview of current heterologous expression systems for xenobiotic metabolising enzymes is given with a special emphasis on the yeast expression system. In a first part, basic properties and relative advantages and drawbacks of each expression system are considered. The second part is dedicated to humanized yeast strains allowing human P450 expression in a tailored redox environment and to the possibility to use such strains to simulate complex metabolisms involving a combination of phase I and phase II reactions. The last part presents how the association of numeric simulation to yeast expression can help in understanding rules controlling metabolic profiles in xenobiotic-acting multienzymatic systems. PMID- 9204414 TI - Ribozymes. PMID- 9204415 TI - Computer-aided calculation of the local folding potential of target RNA and its use for ribozyme design. PMID- 9204416 TI - Computational approaches to the identification of ribozyme target sites. PMID- 9204417 TI - Computer analysis of the conservation and uniqueness of ribozyme-targeted HIV sequences. PMID- 9204418 TI - Selection of accessible sites for ribozymes on large RNA transcripts. PMID- 9204419 TI - Selection of efficient ribozyme cleavage sites in target RNAs. PMID- 9204420 TI - Chemical synthesis, analysis, and purification of ribozymes. PMID- 9204421 TI - A practical method for the production of RNA and ribozymes. PMID- 9204422 TI - Preparation of templates for production of ribozymes and substrates. PMID- 9204423 TI - Enzymatic synthesis and characterization of unmodified ribozymes and substrates. PMID- 9204424 TI - T7 transcript length determination using enzymatic RNA sequencing. PMID- 9204425 TI - Chemical and enzymatic approaches to construct modified RNAs. PMID- 9204426 TI - Applications of modified transcripts. PMID- 9204427 TI - Cloning strategies for catalytic antisense RNAs. PMID- 9204428 TI - PCR-based construction of long hammerhead ribozymes. PMID- 9204429 TI - Design and production of asymmetric hammerhead ribozymes. PMID- 9204430 TI - Minimized hammerhead ribozymes. PMID- 9204431 TI - Design of hairpin ribozymes for in vitro and cellular applications. PMID- 9204432 TI - Design of the hairpin ribozyme for targeting specific RNA sequences. AB - The following steps should be taken when designing the hairpin ribozyme to cleave a specific target sequence: 1. Select a target sequence containing BN*GUC where B is C, G, or U. 2. Select the target sequence in areas least likely to have extensive interfering structure. 3. Design the conventional hairpin ribozyme as shown in Fig. 1, such that it can form a 4 bp helix 2 and helix 1 lengths up to 10 bp. 4. Synthesize this ribozyme from single-stranded DNA templates with a double-stranded T7 promoter. 5. Prepare a series of short substrates capable of forming a range of helix 1 lengths of 5-10 bp. 6. Identify these by direct RNA sequencing. 7. Assay the extent of cleavage of each substrate to identify the optimal length of helix 1. 8. Prepare the hairpin tetraloop ribozyme to determine if catalytic efficiency can be improved. PMID- 9204433 TI - Design and preparation of sequence-specific RNase P ribozymes. PMID- 9204434 TI - Theoretical considerations in measuring reaction parameters. PMID- 9204435 TI - Experimental approaches for measuring reaction parameters. PMID- 9204436 TI - Determination of catalytic parameters for hairpin ribozymes. PMID- 9204437 TI - Characterizing ribozyme cleavage reactions. PMID- 9204438 TI - Defining optimum reaction conditions for hammerhead ribozymes. PMID- 9204439 TI - Using fluorescence resonance energy transfer to investigate hammerhead ribozyme kinetics. PMID- 9204440 TI - Design of hybridizing arms in hammerhead ribozymes. PMID- 9204441 TI - Optimization of hammerhead flanking sequences using oligonucleotide facilitators. PMID- 9204443 TI - Selection of fast-hybridizing complementary RNA species in vitro. PMID- 9204442 TI - Enhancement of ribozyme function by RNA binding proteins. PMID- 9204444 TI - In vitro selection of hairpin ribozymes. PMID- 9204445 TI - The detection of hammerhead ribozyme cleavage by RT-PCR methods. PMID- 9204446 TI - Detection of ribozyme cleavage products using reverse ligation-mediated PCR (RL PCR). PMID- 9204448 TI - Trans-splicing reactions by ribozymes. PMID- 9204447 TI - Quantitation of ribozyme target abundance by QCPCR. PMID- 9204449 TI - Ligation of RNA molecules by the hairpin ribozyme. PMID- 9204450 TI - Mutagenesis and modeling of the hairpin ribozyme family. PMID- 9204451 TI - Preparation of homogeneous ribozyme RNA for crystallization. PMID- 9204452 TI - Establishing suitability of RNA preparations for crystallization. Determination of polydispersity. PMID- 9204453 TI - A sparse matrix approach to crystallizing ribozymes and RNA motifs. PMID- 9204454 TI - Crystallographic analyses of chemically synthesized modified hammerhead RNA sequences as a general approach toward understanding ribozyme structure and function. PMID- 9204455 TI - tRNA delivery systems for ribozymes. PMID- 9204456 TI - Expressing ribozymes in plants. PMID- 9204457 TI - Using microinjection of Xenopus oocytes to express and optimize ribozymes in vivo. PMID- 9204458 TI - Exogenous cellular delivery of ribozymes and ribozyme encoding DNAs. PMID- 9204459 TI - Optimization of lipid-mediated ribozyme delivery to cells in culture. PMID- 9204460 TI - Retroviral delivery and anti-HIV testing of hammerhead ribozymes. PMID- 9204461 TI - Hairpin ribozyme gene therapy for AIDS. PMID- 9204462 TI - Clinical aspects of ribozymes as therapeutics in gene therapy. PMID- 9204463 TI - Diagnosis of lateralized lumbosacral disk herniation with magnetic resonance imaging. AB - A left-lateralized, lumbosacral intervertebral disk herniation, which was not apparent on epidurography, was diagnosed in a dog with magnetic resonance imaging. Precise, preoperative localization and characterization of the lesion allowed surgical approach and excision with minimum disruption of surrounding tissues. PMID- 9204464 TI - Canine spinal nephroblastoma. AB - An eight-month-old, female, mixed-breed dog was presented with bilateral hind limb paralysis that reportedly developed over a two-to-three week period and was not associated with trauma. Plain radiographs of the spinal column were unremarkable, and a myelogram outlined an intramedullary mass of the spinal cord at the first lumbar (L1) vertebra. A hemilaminectomy was performed, and a mass that was identified histologically as nephroblastoma was excised from the spinal cord. Following surgery, the dog became fully ambulatory, and at 22 months postsurgery she remains clinically normal. The diagnosis, treatment, progression, histogenesis, and pathology of canine nephroblastoma are discussed. PMID- 9204465 TI - Spinal tumors in 37 dogs: clinical outcome and long-term survival (1987-1994). AB - The current management of dogs with spinal canal neoplasia in a large veterinary institution was evaluated. Postoperative survival time and prognostic indicators for survival were examined. Spinal neoplasms in dogs and humans also were compared. Thirty-seven cases with histologically confirmed spinal tumors were included in the study. The cervical region was affected most commonly, and 23 (62%) of 37 cases had extradural tumors. A hemilaminectomy or a dorsal laminectomy was performed in each case; three cases received adjuvant treatment. Twelve (32%) cases were euthanized at the time of surgery, and two died immediately after surgery. One dog was euthanized 20 days after surgery because of persistent clinical signs. Twenty-two cases were followed postoperatively; nine different types of primary tumors were confirmed by histological examination of tissue specimens from these 22 cases, and three cases had metastatic lesions. The median survival time of these 22 cases was 240 days. Twelve (32%) of the 37 cases had nerve-sheath tumors; the median survival time for these 12 cases was 180 days. No prognostic indicators were identified. However, median survival times of cases with benign versus malignant tumor types were 1,410 days and 180 days, respectively (p of 0.07). Four cases each had a myxoma/myxosarcoma, a tumor previously unreported in the spinal canal in dogs. PMID- 9204466 TI - Resolution of superficial necrolytic dermatitis following excision of a glucagon secreting pancreatic neoplasm in a dog. AB - An 11-year-old, neutered male standard poodle was diagnosed with superficial necrolytic dermatitis and a glucagon-secreting pancreatic islet neoplasm based on clinical, biochemical, histopathological, immunohistochemical, and hormonal findings. Hyperglucagonemia, hyperinsulinemia, and hypoaminoacidemia were observed on preoperative laboratory analysis. Abnormal laboratory values returned to normal, and complete resolution of skin lesions occurred after tumor excision. The dog has remained clinically normal for six months following surgery. PMID- 9204467 TI - Radiotherapy of incompletely resected, moderately differentiated mast cell tumors in the dog: 37 cases (1989-1993). AB - Thirty-seven dogs with moderately differentiated, cutaneous mast cell tumors had incomplete surgical excisions as determined by histopathology, but no gross evidence of tumor. All dogs were irradiated to a total dose of between 46.2 and 48.0 Gy using either an orthovoltage source (n = 20) or a linear accelerator (megavoltage) (n = 17). Radiation was delivered to an area bordered by margins of 3 cm or greater around the surgical scar. The mast cell tumors had not recurred in 97% of dogs by one year after radiation therapy and had not recurred in 93% of dogs by three years after radiation. Both orthovoltage and megavoltage radiation provide excellent local control of moderately differentiated mast cell tumors in dogs. PMID- 9204468 TI - Metastatic carcinoma presenting as hind-limb lameness: diagnosis by synovial fluid cytology. AB - A dog presented for evaluation of left hind-limb lameness and pain associated with manipulation of the tail. Synovial metastasis of a carcinoma was diagnosed by joint fluid examination. A primary bronchiolar-alveolar carcinoma with widespread (including synovial and skeletal) metastases was diagnosed on postmortem examination. Metastasis to synovial surfaces is uncommon, but when it occurs, the metastasis-induced arthritis may be the initial presenting complaint for which medical attention is sought. Although rarely reported, cytological examination of synovial fluid may be diagnostic. This paper presents an interesting clinical case and reviews the literature concerning metastatic disease of the synovium. PMID- 9204471 TI - Digital pad transposition for replacement of the metacarpal or metatarsal pad in dogs. AB - A technique for digital pad transposition is described and illustrated. This technique has application for use in cases of metacarpal or metatarsal pad neoplasia or severe trauma. The transposed digital pad will provide a weight bearing surface of heavy, keratinized epidermis in cases where the normal metacarpal or metatarsal footpad has been removed. The use of the technique in four clinical cases of footpad neoplasia also is reported. PMID- 9204470 TI - Primary adenocarcinoma of the gland of the nictitating membrane in a cat. AB - An 11-year-old, neutered, male domestic shorthair was presented with a five-month history of recurrent, unilateral, seromucoid discharge from the right eye. A verrucous mass extended from the posterior aspect of the nictitating membrane. Adenocarcinoma of the gland of the nictitating membrane (GNM) was diagnosed upon biopsy. The cat subsequently developed metastases to the lungs, pleura, mediastinum, liver, and kidneys and died six months after clinical signs first were observed. Little is known about the biological behavior of adenocarcinoma of the GNM in cats. This is the first report that describes the natural progression of this disease. PMID- 9204469 TI - Feline cutaneous squamous cell carcinoma of the nasal planum and the pinnae: 61 cases. AB - Cutaneous squamous cell carcinoma is a common tumor in cats and frequently occurs on the nasal planum and the pinnae. The medical records of 61 cats were reviewed for this retrospective study. Typical presentation was an older (median age, 12 years) cat with an erythematous, crusty, and erosive lesion. Methods of treatment included surgery, radiation, and cryotherapy. Disease-free interval and survival time were calculated for each case and grouped according to lesion location and treatment type. All treatments were found to be effective, with surgery resulting in the longest disease-free interval (median, 594 days). PMID- 9204472 TI - Stellate rhytidectomy: superior entropion repair in a dog with excessive facial skin. AB - A four-year-old Chinese shar pei was presented for entropion repair that had not been corrected adequately with two prior Hotz-Celsus procedures. The primary cause for the failure was the weight of the excessive, superior facial folds often found in this breed. A new technique is presented to measure and remove these folds in a stellate pattern by following natural stress lines, thus effecting surgical repair of the superior lid entropion. PMID- 9204474 TI - German shepherd dog pyoderma: a prospective study of 12 dogs. AB - Twelve German shepherd dogs, each diagnosed as having a recurrent or refractory deep pyoderma (i.e., German shepherd dog pyoderma [GSP]), were evaluated for several parameters over a six-year period. Results indicated that GSP could be associated with flea allergy dermatitis, atopic dermatitis, food allergy, cell mediated immunodeficiency, or hypothyroidism, or could be an idiopathic disease. The combination of diseases present for a given dog varied from case to case. Adequate control of the pyoderma was achieved only after each identified underlying disease was treated specifically, along with aggressive concurrent medical therapy using systemic antibiotics and medicated baths. PMID- 9204473 TI - The effect of wound irrigation with bupivacaine on postoperative analgesia of the feline onychectomy patient. AB - Eighteen cats that each underwent an elective onychectomy were evaluated using a double-blind study design to determine if wound irrigation with bupivacaine prior to wound closure would decrease postoperative pain. The cats were divided alternately into an experimental group (n = 9) and a control group (n = 9). The experimental patients received bupivacaine in each incision prior to closure. The control patients received saline in each incision prior to closure. The patients were evaluated for postoperative pain using a pain-score system. The bupivacaine treated patients had a significantly higher mean pain score at two hours following recovery from anesthesia than the saline-treated patients. At three hours following recovery from anesthesia, pain scores were not significantly different. PMID- 9204475 TI - Survival and prognostic factors in 189 dogs with dilated cardiomyopathy. AB - A survival analysis was performed using the case records of 189 dogs, including 38 breeds, with congestive heart failure caused by dilated cardiomyopathy (DCM). Overall prognosis was poor, with survival rates of 17.5% at one year and 7.5% at two years. Prognosis in the individual case of DCM proved to be difficult to predict at the time of initial examination. Only three of 27 tested independent predictors of survival were identified. The most significant predictive variables were age at onset of clinical signs, followed by dyspnea and ascites (as noted on the physical examination). PMID- 9204476 TI - Expression of Tiam-1 in the developing brain suggests a role for the Tiam-1-Rac signaling pathway in cell migration and neurite outgrowth. AB - During development proper neuronal migration and neurite extension are essential for the formation of functional neuronal networks. These processes require the reorganization of the cytoskeleton by modifying the dynamics of actin filaments and microtubules. The Rho subfamily of GTPases regulates actin cytoskeletal changes during development. Tiam-1, a GDP-GTP exchange factor for the small GTPase Rac and implicated in tumor invasion and metastasis, is expressed in the developing CNS. To study the function of Tiam-1 in neuronal migration and neurite extension, we examined the pattern of Tiam-1 expression in weaver mice, in which cerebellar granule cells fail to migrate to their final position and subsequently die. Tiam-1 is expressed in wild-type granule cells as they migrate to the internal granular layer and send axone. In contrast, weaver homozygous animals do not express. Tiam-1 in premigratory granule cells. Heterozygous animals, in which granule cells exhibit a slow rate of migration, express low levels of Tiam-1. In the cerebral cortex, Tiam-1 is also expressed in migrating neurons. Our findings suggest that Tiam-1 contributes to cytoskeletal reorganization required during cell migration and neurite extension in defined neuronal populations, presumably by activation of Rac. PMID- 9204477 TI - Aberrant protein trafficking in Trembler suggests a disease mechanism for hereditary human peripheral neuropathies. AB - The naturally occurring mouse mutant Trembler (Tr) represents an animal model for inherited human neuropathies caused by point mutations affecting peripheral myelin protein 22 (PMP22). We describe the likely pathogenic cellular mechanism underlying the observed myelin deficiency. In Tr/+ animals, PMP22 immunoreactivity was found not only in compact myelin but also abundantly in the cytoplasm of Schwann cells. Based on these observations, the biosynthesis of wildtype and Tr protein was examined in transfected cells. While wildtype PMP22 was readily transported to the plasma membrane, Tr protein was localized mainly in the endoplasmic reticulum. Coexpression revealed a dominant effect of Tr on protein trafficking of wildtype PMP22. In agreement with the findings in vitro, Tr protein was not detectable in myelin of Tr/0 mice. PMID- 9204478 TI - Cloning and expression of a novel murine semaphorin with structural similarity to insect semaphorin I. AB - We describe a novel semaphorin family member, Sema VIa, with 25-36% sequence identity at the amino acid level in the semaphorin domain to previously published mouse homologues. This novel family member shares considerable homology with the best characterized murine semaphorin, Sema III (also known as SemD), at the 5' end but is divergent from Sema III near the 3' end because it contains a putative transmembrane domain. Remarkably, of the known semaphorins, Sema VIa bears the greatest structural similarity to insect Sema I, although it contains a much larger intracellular domain. We propose, therefore, that Sema VIa is the prototype of a new class (class VI) of semaphorins. In order to gain insights into potential functions of Sema VIa, we have compared mRNA expression of Sema VIa to that of Sema III during development. In the nervous system, Sema VIa is expressed in strikingly localized and transient patterns that are markedly different from those of Sema III. Interestingly, Sema VIa and Sema III frequently exhibit complementary or adjacent loci of expression. We suggest that Sema VIa may be important to nervous system development via a mechanism that involves cell cell communication. PMID- 9204479 TI - Susceptibility to cell death induced by mutant SV40 T-antigen correlates with Purkinje neuron functional development. AB - Purkinje cells are uniquely susceptible to a number of physical, chemical, and genetic insults both during development and in the mature state. We have previously shown that when the postmitotic state of murine Purkinje cells is altered by inactivation of the retinoblastoma tumor susceptibility protein (pRb), immature as well as mature Purkinje cells undergo apoptosis. DNA synthesis and neuronal loss are induced in postmitotic Purkinje cells dependent upon the pRb binding portion of SV40 large T antigen (T-ag). In the present study, Purkinje cell targeting of a mutant T-ag, PVU, which does not bind pRb, reveals disparate cerebellar phenotypes dependent upon temporal differences in transgene expression. Strong embryonic and postnatal transgene expression in three lines alters Purkinje cell development and function during the second postnatal week, causing ataxia without Purkinje cell loss. In contrast, two other transgenic lines reveal that PVU T-ag expression following normal Purkinje cell maturation causes rapid Purkinje cell degeneration. The second and third postnatal weeks of cerebellar development, which include the major period of synaptogenesis, appear to be the defining stage for the two PVU-induced phenotypes. These data indicate that Purkinje cell death susceptibility varies with developmental stage. PMID- 9204480 TI - Phenotypic analysis of mice lacking the highly abundant Purkinje cell- and bipolar neuron-specific PCP2 protein. AB - The Purkinje cell protein-2 (Pcp2, also known as L7) gene is abundantly expressed only in Purkinje cells of the cerebellum and bipolar neurons of the retina. The spatio-temporal expression pattern of this gene suggests a role for PCP2 in Purkinje cell development or normal cell physiology. A PCP2-deficient mouse was created by gene targeting to test the hypothesis that it is required for Purkinje cell development or function. Although normally present in abundance, the absence of PCP2 in null animals caused no observable cerebellar abnormalities. Behavioral analysis reveals normal abilities for balance and coordination. Null cerebellum has normal Purkinje cell numbers, morphology, and ultrastructure. Retinal bipolar neurons appear similarly unaffected. Aged null animals (22 months) were also examined and no deficits were detected using the same behavioral and histologic analyses. Although the null animal does not reveal the function of PCP2, it does rule out an essential role for PCP2 in Purkinje cell development, in Purkinje cell survival, and in at least some aspects of cerebellar function. PMID- 9204481 TI - Expression of an L1-related cell adhesion molecule on developing CNS fiber tracts in zebrafish and its functional contribution to axon fasciculation. AB - E587 antigen, an L1-related cell adhesion molecule, is expressed by growing axons and has previously been shown to enhance axon growth and to mediate fasciculation of axons from newborn retinal ganglion cells in goldfish. In zebrafish, the monoclonal antibody E17 against E587 antigen stains all axons in the primary tracts and commissures from 17 h postfertilization (pf) onward and axons which are added subsequently to this scaffold. Moreover, Fab fragments of an E587 antiserum (E587 Fabs) injected into the ventricle of 30-h pf zebrafish embryos caused a marked defasciculation of distinct axon bundles in the posterior commissure, in hindbrain commissures, and in longitudinal tracts of the hindbrain, where they also caused increased crossings between fascicles. The regulated expression of E587 antigen by all developing axons and the effects caused by E587 Fabs show that E587 antigen contributes to the formation of tight and orderly fascicles in the developing CNS. PMID- 9204482 TI - 'Oops!': performance correlates of everyday attentional failures in traumatic brain injured and normal subjects. AB - Insufficient attention to tasks can result in slips of action as automatic, unintended action sequences are triggered inappropriately. Such slips arise in part from deficits in sustained attention, which are particularly likely to happen following frontal lobe and white matter damage in traumatic brain injury (TBI). We present a reliable laboratory paradigm that elicits such slips of action and demonstrates high correlations between the severity of brain damage and relative-reported everyday attention failures in a group of 34 TBI patients. We also demonstrate significant correlations between self- and informant-reported everyday attentional failures and performance on this paradigm in a group of 75 normal controls. The paradigm (the Sustained Attention to Response Task-SART) involves the withholding of key presses to rare (one in nine) targets. Performance on the SART correlates significantly with performance on tests of sustained attention, but not other types of attention, supporting the view that this is indeed a measure of sustained attention. We also show that errors (false presses) on the SART can be predicted by a significant shortening of reaction times in the immediately preceding responses, supporting the view that these errors are a result of 'drift' of controlled processing into automatic responding consequent on impaired sustained attention to task. We also report a highly significant correlation of -0.58 between SART performance and Glasgow Coma Scale Scores in the TBI group. PMID- 9204483 TI - Memory and aphasia. AB - Sixty-one acute and 17 chronic vascular left-hemisphere damaged patients were tested with five memory tasks that investigated verbal short-term (digit span) and long-term (paired-associate and story learning) memory, and spatial short- and long-term memory (Corsi's span and learning). Both brain-damaged groups were significantly impaired in all memory tasks (except for chronic patients in the story learning task) compared to normal controls. The presence of aphasia and locus of lesion (anterior, posterior and deep) had no effect on the memory impairment, with only one exception of paired-associate learning that was better performed by non-aphasic than aphasic patients. Eleven subjects were better at paired-associate learning than story recall, the reverse dissociation was never found. Finally, chronic patients performed significantly better than acute patients only in the Corsi's learning task. PMID- 9204484 TI - Learning and forgetting processes in Parkinson's disease: a model-based approach to disentangling storage, retention and retrieval contributions. AB - Learning and forgetting a prose passage was studied in 20 patients with Parkinson's disease and in 20 normal control subjects by means of stochastic models, with the aim of identifying the learning and retaining abilities that are affected by Parkinson's disease. Results suggested that Parkinson's disease patients are impaired in developing automatic processing both during learning and retaining, while functions that require active attention are spared. The automatic/intentional dissociation, which is the hallmark of motor disturbance in Parkinson's disease, extends to memory abilities, and, on the grounds of neuroanatomical, neurochemical and neurophysiological correlates, suggests that the memory deficit in Parkinson's disease may be contingent on a dysfunction of the medial prefrontal-cingulate cortex. PMID- 9204485 TI - Is memory loss without anatomical damage tantamount to a psychogenic deficit? The case of pure retrograde amnesia. AB - Following a car accident, a patient remained unconscious for approximately 20 min and confused for a few hours. When he could be questioned, he was found to have lost all past memories. The retrograde amnesia covered his whole life and concerned autobiographic events as well as famous facts and encyclopaedic knowledge. It also partially involved the verbal and visual lexicon. Reading, writing and counting were no longer possible. The profound impairment of retrograde memory contrasted with the preservation of anterograde memory, which permitted the patient to reacquire some of the notions he had lost, without, however, recovering the feeling of a personal experience of autobiographical information. Four years later, the retrograde deficit was unmodified, except for what had been relearnt. The search for data in support of an organic or psychological aetiology was negative. No signs of brain damage were apparent at the neurological examination and on CT, MRI and SPECT. On the other hand, there was no evidence of a psychiatric history, psychological stress or emotional precipitants that could substantiate the hypothesis that the patient derived a primary or secondary gain from amnesia. We propose that cases of focal retrograde amnesia, similar to the present one, deserve to be classified separately from organic and psychogenic forms under the label of 'functional' retrograde amnesia, a syndrome in which the threshold of activation of premorbid memories is abnormally raised by the trauma, leaving the encoding and retrieval of new memories unaffected. PMID- 9204486 TI - The phonological short-term store-rehearsal system: patterns of impairment and neural correlates. AB - Two left brain-damaged patients (L.A. and T.O.) with a selective impairment of auditory-verbal span are reported. Patient L.A. was unable to hold auditory verbal material in the phonological store component of short-term memory. His performance was however normal on tasks requiring phonological judgements, which specifically involve the phonological output buffer component of the rehearsal process. He also showed some evidence that rehearsal contributed to the immediate retention of auditory-verbal material. Patient T.O. never made use of the rehearsal process in tasks assessing both immediate retention and the ability to make phonological judgements, but the memory capacity of the phonological short term store was comparatively preserved. These contrasting patterns of impairment suggest that the phonological store component of verbal short-term memory was severely impaired in patient L.A., and spared, at least in part, in patient T.O. The rehearsal process was preserved in L.A., and primarily defective in T.O. The localisation of the lesions in the left hemisphere (L.A.: inferior parietal lobule, superior and middle temporal gyri; T.O.: sub-cortical premotor and rolandic regions, anterior insula) suggests that these two sub-components of phonological short-term memory have discrete anatomical correlates. PMID- 9204487 TI - Can the recognition of living things really be selectively impaired? AB - Brain damage sometimes impairs recognition of living things relative to nonliving things. One interpretation of this dissociation is that recognition of living things depends on specialized mechanisms not used for the recognition of nonliving things; another is that patients have damage to a general purpose system, and recognition of living things taxes this system more heavily. Farah et al., (Neuropsychologia, Vol. 29, pp. 185-193, 1991 [7]) found that a set of general factors did not account for patients' impaired recognition of living things; thus they favored the specialized mechanisms account. The current paper builds on Ref. [7] in two ways. First, other research suggests a number of additional factors that might account for the results in Ref. [7] but were not tested there. Second, statistical methods in Ref. [7] may have implicitly favored the conclusion obtained there, so we use more conservative methods. The current work accounts for all these things, and finds that recognition of living things is still disproportionately impaired. PMID- 9204489 TI - Object-based visual attention in luminance increment detection? AB - A cued reaction time task was used to test the hypothesis that there is an 'object-based' component to shifts of attention mediating the detection of luminance increment targets. The test stimulus consisted of two intersecting triangles forming a 'Star of David'. In two experiments, the cue was a brief brightening of one triangle. The target (a bright green dot) appeared on one of the triangles after a delay of 100, 200 or 500 msec. In one experiment, the target was more likely to appear on the cued triangle. In a second experiment, there was no contingency between cue and target. In both cases, reaction times to targets which appeared outside (but not inside) the cued triangle were more than 10 msec longer than other targets, but only at the shorter cue-target delays. This indicates that the attentional system which regulates luminance increment detection cannot select the cued triangle. It appears that the attentional spotlight can be briefly deformed into a triangular shape, and that it is the rapid, fast-decaying and reflexive exogenous system, rather than the slower acting, persistent and voluntary endogenous system, that mediates this effect. A third experiment using a central, symbolic cue showed no significant cue-validity effects, indicating no contribution from the endogenous system. It is concluded that tasks requiring only stimulus detection cannot unequivocally discriminate between spatial and object-based components of attention. PMID- 9204488 TI - Distractor-dependent frontal neglect. AB - The effect of distractor load on visual search was examined in a patient with visual neglect following infarction of the right frontal lobe. The spatial extent of his left-sided neglect was modified greatly by changing stimulus attributes. When targets were highly discriminable compared to distractors, or distractor density was low, or when the subject was asked to cancel distractors as well as targets, he was able to direct his search to the extreme left of search arrays and there was little or no evidence of neglect. By contrast, similar changes in distractor load had little or no effect on the neglect of a patient with a fronto parietal lesion. These findings suggest that distractability towards ipsilesional stimuli may be an important component of neglect in individuals with only frontal lobe injury. PMID- 9204490 TI - Attention and oculomotor control: a high-density ERP study of the gap effect. AB - In a gap paradigm, healthy adult subjects performed visually triggered saccades to peripheral targets either with the fixation stimulus remaining on (overlap trials) or going off before target onset (gap trials). All subjects showed faster reaction times in the gap trials (the gap effect). High density scalp event related potentials were recorded time-locked to both the target stimuli and the eye movement onset. We observed three neural correlates of the gap effect: (i) a prefrontal positivity that precedes the target presentation which may reflect specific preparatory processes, (ii) an enhancement of the early cortical visual responses (PI) to the peripheral target in the gap trials, and (iii) a prolongation of parietal activity in the overlap trials relative to the gap trials prior to the saccade execution. These results suggest that several factors contribute to the gap effect, each having its own neural basis. PMID- 9204491 TI - Attentional competition between modalities: extinction between touch and vision after right hemisphere damage. AB - The phenomenon of extinction, which occurs frequently after unilateral brain damage, involves a failure to detect the more contralesional of two simultaneously presented stimuli, but with preserved detection of single ipsilesional or contralesional stimuli. Current accounts suggest that the disorder reflects a bias of selective attention, in which ipsilesional stimuli have a competitive advantage. Extinction may be manifested within any one of the major sensory modalities (vision, audition, touch), or it may occur within several modalities in a given individual. Given recent evidence in normals for attentional links between separate sensory modalities, we examined whether extinction can also occur cross-modally, i.e. for double-simultaneous stimuli in separate sensory modalities. We tested whether an ipsilesional event sufficient to extinguish a contralesional stimulus within the same modality may also extinguish a contralesional stimulus in a different modality. Our three patients had right hemisphere damage, and reliable within-modality extinction for visual and tactile stimuli. They also showed significant cross-modal extinction, such that an ipsilesional tactile (or visual) event extinguished awareness of a simultaneous visual (or tactile) event on the contralesional side. These results, which provide the first quantitative evidence for cross-modal extinction, were replicated in a second experiment in which visual and tactile stimuli in the cross-modal conditions were presented at non-homologous elevations within each hemispace. We conclude that after unilateral damage, ipsilesional stimuli have a competitive advantage over contralesional stimuli, and that this affects competition between stimuli from different modalities as well as stimuli within the same modality. These findings are consistent with recent evidence for competitive interactions between tactile and visual events in the control of spatial attention in normals. PMID- 9204492 TI - Remote memory in patients with acute brain injuries. AB - Remote memory was investigated in an unselected sample of 26 patients with either unilateral tumours in the temporal lobes or traumatic brain injuries. Six patients underwent excisions within the left temporal lobe, and nine patients were operated on within the right temporal lobe. In both groups, patients with excisions including and sparing the hippocampal formation were studied. Their performance was compared to that of 11 patients with moderate to severe head trauma and to a normative sample of 214 healthy controls. Remote memory was assessed using a famous events test with items of extremely low salience that had been proven to be of low difficulty for those old enough at the time of the event's actuality. The results show severely disturbed retrograde memory functions in the left temporal tumour group. These patients achieved similar scores to patients with severe traumatic brain injury. Right hemispheric patients showed a pattern of results comparable to that of healthy controls. The strongest effects were in the free recall part of the test. In most of the patients, no graded memory loss was observable. No consistent association to recent memory function could be identified. Since most of the remote memory test items used denoted famous names which were cued by rich semantic information, the type of deficit seen may be best understood in terms of a specific dysfunction of the semantic stores containing information about famous proper names. PMID- 9204493 TI - Patterns of oculomotor scanning in patients with unilateral posterior parietal or frontal lobe damage. AB - Eye movements were recorded during the inspection of dot patterns in control subjects and in patients with acquired unilateral brain damage involving posterior parietal or frontal cortical regions. Normal subjects adapted their oculomotor scanning pattern effectively to the stimulus configuration. Patients' oculomotor scanning patterns were characterized by a rather rigid sequence of fixations and saccades, with no evidence for a systematic and flexible spatio temporal organization. In a stimulus condition where dots were grouped, patients with frontal damage accurately shifted their gaze between the dot groups, but had difficulties with dot sampling. These observations suggest that the posterior parietal damage mainly affected the visuo-spatial guidance of the scanpath, whereas the frontal damage impaired its planning. It is concluded that both, posterior parietal and frontal brain structures, and their reciprocal connections, are part of a distributed neural network subserving visually-guided oculomotor scanning, and that the spatio-temporal organization of the scanpath depends critically on both structures and their close co-operation. PMID- 9204494 TI - The selective breakdown of frontal functions in patients with obsessive compulsive disorder and in patients with schizophrenia: a double dissociation experimental finding. AB - In an our recent preliminary study, we reported the neuropsychological finding of a double dissociation in the frontal lobe functioning between 25 OCD patients and 25 schizophrenics. The first group performed normally in the Wisconsin Card Sorting Test (WCST), which is considered sensitive to Dorso-Lateral Prefrontal Cortex (DLPFC) dysfunctions and abnormally to the Object Alternation Test (OAT), which has been proposed as a tool sensitive to Orbito-Frontal Cortex (OFC); on the other hand, schizophrenics performed abnormally to the WCST and normally to the OAT. The present study, conducted on a new sample of 60 schizophrenic in patients, 60 OCD in-patients and 30 normal subjects, matched according to age, educational level, handedness and duration of illness, confirms our preliminary data and it suggests a more selective impairment of OFC system in OCD and of DLPFC in schizophrenia. Moreover, schizophrenic patients with paranoid subtype showed worse WCST performance compared to non-paranoid subtype. Our results could open some interesting perspectives about the neuroanatomical systems involved in these two major psychiatric illnesses and so, about their pharmacological treatment, on the basis of the prominent catecholaminergic characterization of the DLPFC and, respectively, the cholinergic innervation of the OFC. PMID- 9204495 TI - Interaction between the hemispheres in split-brain cats. AB - Functional interactions between the two hemispheres were studied in adult split brain cats. The aims were to assess whether monocular learning developed independently or that there were clues for interactions between the two sides of the brain during acquisition of opposite learning tasks. Experimental cats learned two visual pattern discriminations in which one pattern was positive for the right eye, whereas the other pattern was positive for the left eye. Control cats learned the same problems, but the same pattern was positive for both eyes. The open eye was changed from one session to the next in both groups of cats. In general, monocular performances of experimental cats were asymmetrical because they learned better and faster with one eye than with the other eye. Instead, no differences between the eyes were found in control cats. Statistical analysis of the data indicated that learning in experimental cats was significantly slower than learning in control cats, and that the difference between monocular performances was significantly greater for the experimental group than for the control group. The slower and asymmetrical monocular learning of experimental cats may reflect a conflict and a competition between the hemispheres for the control of learning behaviour, resulting in the dominance of one of them. Thus, some information about the stimuli must have been transmitted via the remaining interhemispheric connections. Symmetrical monocular learning of control group indicated that the competition for the control of behaviour was not present because there was no conflict between the hemispheres. PMID- 9204496 TI - Lexicon size and its relation to foot preference in the African grey parrot (Psittacus erithacus). AB - To study footedness in parrots, an international survey of parrot owners was conducted. Responses were obtained from 524 individuals, including 70 owners of African Grey parrots (all animals > or = 10 months old). All respondents were given a 10-item questionnaire and a standard method for testing foot preference in their pets, and they were asked to count the number of separate words in their pets' lexicons of human speech sounds. Right-footed African Greys (N = 36) had significantly larger lexicons than left-footed African Greys (N = 34; P = 0.01). This difference could not be accounted for by group differences in training efforts or socialization/housing with conspecifics. A non-significant trend in the same direction was found in a comparison sample of Amazon parrots, although these genera are less adept than African Greys at learning human speech sounds. Other investigators have provided convincing evidence of lateralization, in the avian brain, for the analysis and memory of differing types of stimuli. In addition, there appears to be preferential left hypserstriatal activation for long-term memory consolidation. Our results suggest a relationship between lateral asymmetry for motor preference and asymmetric CNS mediation of a 'higher cognitive' function (i.e. the categorization and long-term mnestic processing of human speech sound. PMID- 9204497 TI - Acquisition of protective immunity in Geochelone pardalis against Amblyomma marmoreum (Acari:Ixodidae) nymphal ticks. AB - Significant increases in serum globulins (alpha 1, alpha 2, beta and gamma) were observed in mountain leopard tortoises (Geochelone pardalis) after they had been immunized with nymphal homogenates of Amblyomma marmoreum. There was a concomitant significant increase in the numbers of leukocytes (lymphocytes, basophils, monocytes and eosinophils). Resistance to nymphal-challenge infestations was manifested by reduced feeding time, lower engorgement masses, and significantly fewer (P < 0.0001) numbers of nymphs that moulted. These findings are contrary to the generally reported phenomenon, that ticks do not induce resistance in their natural hosts. PMID- 9204498 TI - A checklist of helminths from the respiratory system and gastrointestinal tracts of African Anatidae. AB - A literature survey revealed that 72 helminth species, including 14 known only to the generic level, had been reported from the digestive or respiratory tracts of 28 species of Anatidae in Africa. Most of the digeneans and nematodes reported, were cosmopolitan species that occur in a range of hosts. However, two groups of cestodes, one consisting of cosmopolitan or Eurasian species and the other consisting of species restricted mainly to sub-Saharan Africa, were apparent. A host-parasite list and a detailed parasite-host list provide the synyonomies related to African records, the host and geographical distribution of each species, and the authority and country of origin for each record. PMID- 9204500 TI - Ulcerative pododermatitis in free-ranging African elephant (Loxodonta africana) in the Kruger National Park. AB - The occurrence of severe lameness in adult African elephant bulls in a shrub Mopane (Colophospermum mopane) ecosystem was investigated. Large ulcers in the soles of at least one front foot were seen in each of the recorded cases. Microscopically, the lesion can be described as a severe, chronic-active, ulcerative, bacterial pododermatitis (complicated by hypersensitivity/septic vasculitis). A variety of bacteria were isolated from these lesions as well as from regional lymph nodes. Streptococcus agalactiae was the most consistent isolate, while Dichelobacter nodosus, the only organism known to be involved with foot disease in domestic ruminants, was isolated from two cases. Contributory factors such as body mass, portal of entry and origin of potential pathogens may have predisposed to the development of the lesions. PMID- 9204501 TI - Camallanus polypteri n. sp. (Nematoda:Camallanidae) in freshwater fishes from Burkina Faso. AB - Camallanus polypteri n. sp.is described from Polypterus bichir (type host), Synodontis schall and Clarias anguillaris in Lake Tingrela, Burkina Faso. It differs from the other African species of the subfamily Camallaninae parasitizing freshwater fishes, in that it lacks tridents. All species of the subfamily Camallaninae that lack tridents differ from the new species by at least one of the two following characters: a greater number of longitudinal ridges on the inner surface of the buccal capsule; and a muscular oesophagus shorter than the glandular oesophagus. In accordance with the classification of Petter (1979), the new species is placed in the genus Camallanus Railliet & Henry, 1915, owing to the length of the posterior chamber of the buccal capsule, which is less than one third of the length of the anterior chamber. PMID- 9204499 TI - Failure to establish chronic infection of the reproductive tract of the male horse with a South African asinine strain of equine arteritis virus (EAV). AB - Eight sexually mature horse stallions were inoculated intranasally with a South African asinine strain of EAV, a strain that was isolated from the semen of a donkey carrier. All horses developed fever, with maximum rectal temperatures of 38.9-39.9 degrees C recorded 3-6 d post challenge. Six horses showed very mild clinical signs of equine viral arteritis and two were asymptomatic. The virus was recovered from the nasopharynxes of six horses 2-7 d after inoculation, and from buffy-coat samples of all horses, 2-11 d after inoculation. Seroconversion to EAV was detected on days 8 and 10 and peak serum-virus-neutralizing antibody titres ranging from log10 1.2-1.8, on days 14-20 after challenge. The titres varied from log10 0.9-1.2 after about 10 weeks, when the experiment was terminated. In three stallions euthanased on days 5, 7 and 9 after challenge, virus was detected inconsistently in different parts of the reproductive tract and urine. No virus was isolated from the tissues of the reproductive tract collected from stallions on days 16, 23 and 68 after challenge. Five stallions were bred to six seronegative mares between 13 and 34 d post challenge. No clinical signs of EAV were observed, and neither was seroconversion detected in any of the mares after mating. No virus was recovered from semen samples collected at the time of breeding. The results of this study demonstrated that the tissues of the reproductive tracts of the stallions did not become persistently infected with a South African asinine strain of EAV. PMID- 9204502 TI - Aspects of rabies epidemiology in Tsumkwe District, Namibia. AB - Aspects of rabies epidemiology were investigated in the Tsumkwe District, Namibia, during December 1993 and January 1994. A cross-sectional seroepidemiological survey for rabies antibodies was carried out in domestic (n = 70) and wild dogs [Lycaon pictus (n = 6)]. An overall seroprevalence rate of 30% was found in domestic dogs, but it must be borne in mind that seroconversion can result from infections from either rabies or rabies-related viruses. Older dogs were more likely to be seropositive and there was spatial and temporal clustering of seropositivity. No wild dogs were found seropositive. A demographic survey of the domestic-dog population in the area showed that the total dog-population size was 132, or 0.027 dogs per km2. The dog population consisted mainly of young dogs with a median age of 1.5 years, and had a female bias of 0.63 males per female. Questionnaire surveys suggested that spotted hyaenas (Crocuta crocuta) and black backed jackals (Canis mesomelas) were the most common larger carnivores found in and around villages, and that dogs were kept mainly for guarding. PMID- 9204503 TI - A scanning electron-microscope examination of the scolex of Houttuynia struthionis. AB - A scanning electron-microscope examination of the scolex of Houttuynia struthionis, a cestode of ostriches, was undertaken in order to study its surface structure. The scolex differs from those of other subfamilies in the family Davaineidae in that it does not have scale-like spines covering the base of the rostellum. Instead, the base is covered with small hooks resembling the larger rostellar hooks in shape. PMID- 9204504 TI - In vitro cultivation of Babesia equi: detection of carrier animals and isolation of parasites. AB - By means of an in vitro culture technique, 75 samples of horse blood were examined for Babesia equi, a causative agent of equine piroplasmosis. At the time of culture initiation, 15 samples were microscopically positive for B. equi, and this was subsequently confirmed by culture diagnosis. Sixty samples showed no parasites in Giemsa-stained thin blood smears. However, after the culturing process, parasites were found in blood smears of 36 of these samples. The sensitivity of the in vitro culture method was such that 2.5 microliters (1/40 of the usual volume used for the above-mentioned samples) of packed erythrocytes obtained from a carrier horse still yielded positive results after cultivation. Cultures were initiated from blood samples stored for up to 120 h at 8 degrees C in vacuum tubes containing EDTA as anticoagulant. These results show that the in vitro culture method is highly sensitive. It can be used to identify B. equi carrier horses, to evaluate the effects of chemotherapeutic intervention, and to isolate field strains of B. equi for further characterization. PMID- 9204505 TI - Effects of growth conditions and incubation times on the expression of antigens of Haemophilus paragallinarum which are detected by monoclonal antibodies. AB - Haemophilus paragallinarum causes infectious coryza in poultry, and a panel of monoclonal antibodies (Mabs) were established, which detect surface antigens of this bacterium. It was postulated that these Mabs could be used to detect antigenic differences between strains of H. paragallinarum used in infectious coryza (IC) vaccines, and isolates made from the field, from poultry vaccinated against IC. It has previously been reported that in South Africa there are three different Mab patterns that have been common to H. paragallinarum isolates for the past three decades. The effects of different growth conditions such as duration of incubation, inoculum size, levels of NAD or NaCl in the medium, and the pH of the medium on these Mab patterns were investigated. It was found that many different factors appear to influence the expression of the antigens detected by the panel of Mabs. It was found that at different stages during the growth cycles, the isolates could be classified into different Mab groups. It was also found that alteration of the inoculum size resulted in Mab-pattern switches. Addition of extra NaCl to the medium, in order to slow the growth rate, was found to result in Mab-pattern switches. pH was found to have significant effects on the levels of expression of the antigens detected by the Mabs, although these changes did not result in Mab-pattern switches. The effects of pH were also found to be highly strain dependent. The use of NAD, rather than sterile chicken serum, in the medium did not significantly alter the levels of expression of these antigens. Alterations of the growth conditions greatly affected the levels of expression of the antigens detected by the Mabs, and were highly strain dependent. It was not possible to predict the effects of a particular growth condition on a particular strain or isolate of H. paragallinarum. PMID- 9204506 TI - Schistosoma mattheei infection in cattle: the course of the intestinal syndrome, and an estimate of the lethal dose of cercariae. AB - Three groups of young oxen were infected percutaneously with cercariae of Schistosoma mattheei. Three of five oxen infected with 248 cercariae kg-1 mass died or were killed in extremis 58-70 d after infection, a fourth survived extremely severe clinical schistosomosis and the fifth was only slightly affected. None of seven calves infected with 187 cercariae kg-1 died, while one of seven exposed to 119 cercariae kg-1 was in extremis (possibly not from schistosomosis) when killed after 378 d. The LD50 appears to be in the region of the highest dose tested (248 cercariae kg-1), but depends on variations in the viability of the cercariae used. The clinical syndrome was characterized by a drastic, rapid loss in body mass; a severe diarrhoea containing blood clots; straining, gnashing of the teeth, occasional groaning, and other signs of abdominal pain; and markedly sunken eyes. Lethally infected oxen did not become recumbent until shortly before death. Some severely affected animals made remarkable, but slow, recoveries without treatment. Schistosomes, in close association with granulomata, are described-apparently for the first time-in the omental veins of cattle. Mean worm development in three calves that died or were killed in extremis in the acute stage of the disease, was 55.5%. In contrast to most previous findings with S. mattheei, in two of these animals, more female than male worms developed. The worms were recovered by perfusion and, in one animal, a large number of intestinal veins were dissected open to estimate the efficiency of the perfusion method. Only 1.9% of the total worm burden had not been removed by perfusion in this animal. PMID- 9204507 TI - Canine visceral leishmaniosis: first case in Zambia. PMID- 9204508 TI - Discovery of a Culicoides imicola-free zone in South Africa: preliminary notes and potential significance. AB - In December 1993, a light-trap survey was made of the Culicoides found at eight horse stables and dairies in the sandy dune field west of Port Elizabeth, South Africa. While it was notable that Culicoides numbers were low (4749) and that the diversity was poor (15 species), the most remarkable fact to emerge, was that C. imicola, the only proven vector of the virus of African horse sickness (AHS), was entirely absent. Though not abundant, C. bolitinos, a sister species of C. imicola, was overwhelmingly dominant (91.7%). Its larvae and pupae develop exclusively in the dung of cattle, but it is a species that is not implicated in the transmission of animal viruses. Elsewhere in South Africa, a frost-free climate, good rainfall and a plentiful supply of livestock would normally lead to the development of large foci of C. imicola. That this is not the case in the Port Elizabeth (P.E.) area is most likely owing to the winds inhibiting adult flight and the sandy soils being nutrient-poor and too well-drained to sustain Culicoides larvae. Studies are needed to confirm that sandy soils cannot sustain C. imicola. If so, the sandy coastal areas hold promise for quarantining against AHS. PMID- 9204509 TI - Regional atrophy of the corpus callosum in subjects with Alzheimer's disease and multi-infarct dementia. AB - Regional areas of the corpus callosum (CC) were evaluated in subjects with dementia. The area of the CC and seven distinct subdivisions were measured in subjects with Alzheimer's disease (AD) (n = 162), multi-infarct dementia (MID) (n = 28), and in a healthy comparison group (n = 36). There were significant differences in the regional areas of the CC for both the AD and MID patients relative to values for the comparison subjects. When mildly demented (MMSE > or = 21), subjects with AD had significantly smaller posterior midbody, isthmus, and splenium and subjects with MID had significantly smaller genu relative to comparison subjects. This study reports different patterns of regional CC area loss in subjects with AD and MID. PMID- 9204510 TI - Basal ganglia volume in adults with Down syndrome. AB - This study was designed to determine the effects of aging on the volume of the basal ganglia in individuals with Down syndrome (DS) and to examine the relationship between basal ganglia volumes, neuropsychological test performance, and dementia status in this population. Subjects were 32 adults with DS. Basal ganglia volumes from 22 of these subjects were compared with those of 22 cognitively-normal individuals, who were individually matched on age, sex, and race. Performance on neuropsychological tests was correlated with basal ganglia volumes for 32 individuals with DS, and basal ganglia volumes of five demented DS subjects were compared with those of 14 non-demented DS subjects. Results indicated larger putamen volumes in the DS subjects, despite significantly smaller total brain volumes. Volumes of caudate and globus pallidus did not differ between DS and control subjects. Although there were some significant correlations between basal ganglia volumes and age, neuropsychological test performance, and dementia status in the DS subjects, these associations appeared to be a reflection of neurodevelopmental or atrophic reductions in overall brain volume rather than a reflection of specific basal ganglia abnormality. Correlations between age and volumes of basal ganglia and total brain were not significantly greater in non-demented DS subjects than in control subjects. Results suggest that volume reductions of the basal ganglia are not a salient feature of aging or of the dementia associated with DS. PMID- 9204511 TI - Brain glucose metabolism in anorexia nervosa and affective disorders: influence of weight loss or depressive symptomatology. AB - Relationships between eating and affective disorders remain complex and unclear. Brain glucose metabolism of anorectic patients has been demonstrated to be reduced both globally and regionally, with a particular relative hypometabolism in the parietal cortex. To explore the possible influence of weight loss or depressive symptomatology on brain metabolism, we studied age- and sex-matched low-weight anorectic and depressed patients, normal-weight depressed patients, and healthy volunteers. Absolute global and regional glucose activity levels were reduced in low-weight patients, with the lowest values being found for anorectic patients. In relative values, anorectic patients showed a significant parietal hypometabolism in comparison to control subjects while they had higher metabolism in the caudate nuclei when compared with the other groups. Absolute hypometabolism of glucose seems to be a consequence of low weight as it was found in both low-weight anorectic and low-weight depressive patients. In addition, absolute glucose values were significantly correlated with body mass index in all subjects. Future positron emission tomographic studies in psychiatric patients should control for alimentary parameters. PMID- 9204512 TI - Intracranial and extracranial blood flow during acute anxiety. AB - While large numbers of studies are available on anxiety and cerebral blood flow (CBF), little is known about their relationship to extracranial (forehead) flow. The participants were 24 generalized anxiety disorder (GAD) patients and 26 normal volunteers. A randomized, between groups, repeated measures design was used to evaluate changes in cerebral blood flow. Measurements of CBF, forehead skin perfusion and ratings of anxiety and physiologic indices were made under resting conditions and during anxiety induction with epinephrine or saline infusions, given under double-blind conditions while subjects inhaled room air or 5% CO2. These subjects were divided into three groups; those with decreased anxiety, those with mild anxiety, and those with more severe anxiety increase. Subjects with severe anxiety showed least hypercarbic CBF increase (indicating cerebral vasoconstriction) and maximal increase in forehead skin perfusion. Those with minimal anxiety had most hypercarbic cerebral vasodilation and least increase in forehead skin perfusion. Forehead skin perfusion correlated positively with anxiety levels, and negatively with hypercarbic cerebral vasodilation. In animals, sympathetic activation limits hypercapnic cerebral vasodilation. Thus, the restricted hypercapnic cerebral vasodilation during severe anxiety may be mediated through cervical sympathetic fibers which innervate cerebral vessels. PMID- 9204513 TI - Dipole source analysis of P300 component of the auditory evoked potential: a methodological advance? AB - A dipole model of the P300 component of the auditory evoked potential was developed with the aim of separating the overlapping P300 subcomponents and assessing the reliability of P300 dipole parameters compared with other methods such as principal component analysis (PCA). P300 was recorded within an auditory oddball paradigm in 57 healthy subjects. Dipole source analysis was performed with Brain Electrical Source Analysis (BESA). With a temporo-basal and a temporo superior dipole per hemisphere, most of the variance of scalp-recorded P300 activity can be explained. When dipole source analysis is used, the test-retest reliability of P300-amplitude measurements is enhanced as compared with PCA or single channel analysis. The P300 activity of the temporo-basal dipoles and that of the temporo-superior dipoles differ in several respects (e.g. latency, correlations with age) and thus appear to reflect functionally different neuronal processes. In spite of problems in the interpretation of dipole locations, it is concluded that dipole source analysis enhances both the reliability and physiological validity of P300 parameters. PMID- 9204514 TI - Impaired episodic memory retrieval in a case of probable psychogenic amnesia. AB - A patient with severe, selective retrograde amnesia for personal material diagnosed as probable psychogenic amnesia, was investigated intensively neuropsychologically with cranial computed tomography (CCT), magnetic resonance imaging (MRI), and single photon emission tomography (SPECT). The patient was of average intelligence and memory with no anterograde amnesia. No evidence for structural brain damage was detected in CCT and MRI. SPECT, performed about 3 weeks after the onset of symptoms, demonstrated reduced perfusion in right temporal and frontal areas, that is, in areas which have been suggested as critical for episodic memory retrieval. To study episodic memory retrieval, positron-emission-tomography (PET) blood flow (rCBF) measurements were performed 6 months after the onset of symptoms. During episodic memory retrieval bilateral neuronal activations were observed in the precuneus, the lateral parietal and the right dorsolateral and polar prefrontal cortex. Compared to the results of previous functional imaging studies on episodic memory retrieval, our findings suggest an underlying functional disturbance of brain areas previously demonstrated to be involved in episodic memory retrieval. PMID- 9204515 TI - Hybrid process for the conversion of lignocellulosic materials. AB - Because of the recalcitrant nature of lignocellulosic materials, it is important to pretreat the biomass in order to obtain a suitable material for the bioconversion. In this study, two different types of pretreatments were performed. The first experiment used a 2-gal Parr reactor operated at 140, 150, 160, and 170 degrees C with sulfuric acid concentrations varying from 0.5 to 2%. A second pretreatment was performed with a two-stage low-temperature process. The first-stage pretreatment was performed at 100 or 120 degrees C with sulfuric acid concentrations of 0.5, 2, and 5% followed by a second-stage pretreatment at 120 degrees C with 2% acid concentration. The best residues for enzymatic hydrolysis and simultaneous saccharification and fermentations (SSF) came from the higher temperature pretreatment with the Parr reactor. However, a large portion of the xylose fraction was degraded to furfural and glucose was degraded to HMF. On the contrary, the two-stage low temperature pretreatment resulted in a very low percentage of xylose degradation, and no glucose degradation. The residues from this two-stage pretreatment performed satisfactorily toward the production of ethanol by SSFs. This study discusses the results obtained from these experiments. PMID- 9204516 TI - Synthesis of isopropyl-1-thio-beta-D-glucopyranoside (IPTGlc), an inducer of Aspergillus niger B1 beta-glucosidase production. AB - Production of beta-glucosidase in Aspergillus niger B1 is subjected to catabolic repression by glucose. Aspergillus niger B1 grown on bran as a carbon source secreted beta-glucosidase. The maximum level of the enzyme was reached after 7 d of fermentation. Addition of 1% glucose to the medium suppressed beta-glucosidase production to undetectable levels. In this study, the organic synthesis of a potential inducer of beta-glucosidase production by A. niger B1's reported. Isopropyl-1-thio-beta-D-glucopyranoside (IPTGlc) was synthesized using a two-step organic synthesis protocol. The H-NMR data agreed with those reported previously for the galactoside analog. When IPTGlc was added 24 h after inoculation at a final concentration of 0.4 mM, similar levels of beta-glucosidase were reached 3 to 4 d earlier as compared to fermentation without IPTGlc induction. In practice, this may translate to a more efficient method of producing beta-glucosidase from this fungus. PMID- 9204517 TI - The effect of formaldehyde on the growth of Candida diddensii 74-10 and Candida tropicalis R-70. AB - The effects of formaldehyde on the growth of two strains of fodder yeasts Candida diddensii 74-10 and Candida tropicalis R-70 were investigated using the method of continuous cultivation under conditions of carbon limitation and at dilution rates of 0.1/h and 0.25/h. The results indicate that formaldehyde induces a decrease in the yield of biomass, but stimulates the synthesis of protein and RNA. The authors studied the activities of the following enzymes: NADPH-linked glutamate dehydrogenase, NADH-linked glutamate dehydrogenase, alanine dehydrogenase, glutamine synthetase, and glutamate synthase, which are utilized in nitrogen metabolism. The data obtained showed an increase in the activity of the glutamate dehydrogenase pathway of ammonium assimilation. It was also established that formaldehyde caused considerable changes in the micro-organisms at the higher dilution rate. PMID- 9204518 TI - Isotherms for adsorption of cellobiohydrolase I and II from Trichoderma reesei on microcrystalline cellulose. AB - Adsorption to microcrystalline cellulose (Avicel) of pure cellobiohydrolase I and II (CBH I and CBH II) from Trichoderma reesei has been studied. Adsorption isotherms of the enzymes were measured at 4 degrees C using CBH I and CBH II alone and in reconstituted equimolar mixtures. Several models (Langmuir, Freundlich, Temkin, Jovanovic) were tested to describe the experimental adsorption isotherms. The isotherms did not follow the basic (one site) Langmuir equation that has often been used to describe adsorption isotherms of cellulases; correlation coefficients (R2) were only 0.926 and 0.947, for CBH I and II, respectively. The experimental isotherms were best described by a model of Langmuir type with two adsorption sites and by a combined Langmuir-Freundlich model (analogous to the Hill equation); using these models the correlation coefficients were in most cases higher than 0.995. Apparent binding parameters derived from the two sites Langmuir model indicated stronger binding of CBH II compared to CBH I; the distribution coefficients were 20.7 and 3.7 L/g for the two enzymes, respectively. The binding capacity, on the other hand, was higher for CBH I, 1.0 mumol (67 mg) per gram Avicel, compared to 0.57 mumol/g (30 mg/g) for CBH II. The isotherms when analyzed with the combined Langmuir-Freundlich model indicated presence of unequal binding sites on cellulose and/or negative cooperatively in the binding of the enzyme molecules. PMID- 9204519 TI - Growth of Schizosaccharomyces pombe on glucose-malate mixtures in continuous cell recycle cultures. Kinetics of substrate utilization. AB - The aerobic growth of Schizosaccharomyces pombe on mixtures of glucose and malate was investigated during continuous high cell density cultures with partial cell recycle using a membrane bioreactor. Determination of the specific metabolic rates relative to substrates and products allowed the capacity of the yeast to metabolize malic acid under both oxidative metabolism (carbon limited cultures) and oxidofermentative metabolism (carbon sufficient cultures) situations to be characterized. Under carbon limiting conditions, the specific rate of malate utilization was dependent on the residual concentration and a limit for a purely oxidative breakdown without ethanol formation was observed for a characteristic ratio between the rates of substrate consumption qM/qG of 1.63 g.g-1. In addition, the mass balance analysis revealed the incorporation of malic acid into biomass. In carbon excess environments, the specific rate of malate utilization was dependent on both the residual malate and the specific rate of glucose consumption indicating that in addition to its conversion into ethanol malate can be respiratively metabolized for qM/qG ratios higher than 0.4 g.g-1. PMID- 9204520 TI - Prediction of folding stability and degradability of the de novo designed protein MB-1 in cow rumen. AB - The authors have recently reported on the design of a protein (MB-1) enriched in methionine, threonine, lysine, and leucine. The protein is intended to be produced by rumen bacteria, in a way that would provide high producing lactating cows with limiting amino acids. In this report, MB-1 stability in the rumen is assessed, i.e., where the protein might be found after cell lysis or after being secreted by rumen bacteria. Current in vitro methods used to predict proteolytic degradability in the rumen were used for MB-1, as well as other natural proteins for comparison. MB-1 was found to be more susceptible to degradation than cytochrome c and ribonuclease A. Data indicate that MB-1 will be rapidly degraded if exposed to the rumen environment without protection. The contribution of folding stability to proteolytic stability was also examined. Rumen liquor components were selected to formulate a solution compatible with constraints of thermal denaturation studies. Denaturation curves show that the natural proteins were folded at rumen temperature. The MB-1 denaturation curves indicated that MB 1 does not unfold in a cooperative transition when heated from 20 to 70 degrees C. This suggests that MB-1 structure may be progressively modified as temperature increases, and that a continuum of conformations are available to MB-1. At 39 degrees C, a significant (50%) portion of MB-1 molecules had their tertiary structure unfolded, contributing to proteolytic degradability. Despite the unusual constraints used in MB-1 design (i.e., a maximized content in selected essential amino acids), results show that MB-1 has structural properties similar to previously reported de novo designed proteins. PMID- 9204521 TI - The influence of temperature and alkaline pH on the labeling of free and conjugated MAG3 with technetium-99m. AB - Benzoyl-protected mercaptoacetyltriglycine (S-benzoyl MAG3) is radiolabeled by tartrate transchelation at elevated temperatures or basic pH. The object of this investigation was to establish whether the same 99mTc labeled species are formed when S-acetyl (S-acetyl MAG3)-conjugated compounds are radiolabeled by tartrate transchelation at ambient temperature and neutral pH in contrast to labeling at 95 degrees C and pH 11. S-acetyl MAG3 was conjugated to biocytin and to the amine derivitized oligomers DNA and PNA. Along with free S-acetyl MAG3, these were radiolabeled under the different conditions. Although labeling efficiencies were always lower when labeled at ambient temperature and neutral pH relative to labeling at 95 degrees C or pH 11 (free S-acetyl MAG3 could not be labeled at all), size exclusion and reverse phase HPLC showed no difference with labeling conditions in the radiochemical profiles for labeled DNA and biocytin. In the case of DNA, a cysteine challenge also failed to demonstrate a difference. However, in the case of PNA, some important differences were observed in the size exclusion HPLC radiochromatograms. In addition, PNA labeled at ambient temperature and neutral pH was less stable to transchelation to cysteine. In conclusion, S-acetyl MAG3 conjugated compounds may be radiolabeled at ambient temperature and neutral pH. In most cases, the radiochemical species produced appear to be identical to those formed when labeling is accomplished at 95 degrees C or pH 11. PMID- 9204522 TI - Rapid analysis of 226Ra in waters by gamma-ray spectrometry. AB - Techniques for the determination of 226Ra in waters by gamma-ray spectrometry are examined, with an emphasis on methods capable of completion within 1-3 days of sample collection. Methods discussed utilise either: (i) the 186 keV 226Ra gamma ray, with the contribution from 235U being subtracted after a separate uranium determination, or (ii) measurement of ingrowing 222Rn progeny. An analysis of the statistical and systematic errors associated with each technique is presented. Examples are given of their application to analysis of uranium mine process waters. PMID- 9204523 TI - Radiochromic film dosimetry for clinical proton beams. AB - Depth doses and lateral profiles for proton beams with energies of 100-250 MeV were measured with a high-sensitivity GafChromic MD-55 film, which requires no post-irradiation development. The exposed MD-55 films were evaluated with the RIT 113 film dosimetry system. Depth doses measured with MD-55 film were compared with those obtained with a plane-parallel ionization chamber. The GafChromic film was found to be less sensitive at the distal edge for both modulated and unmodulated beams compared with an ionization chamber, however, the difference in penetration of the beam at the depth of 80-50% distal fall-off measured with this film and the ionization chamber is within 0.1-0.2 mm for studied beams. The lateral beam profiles measured with MD-55 film were in a good agreement with profiles obtained from Kodak XV-2 radiographic film, scanned with the Wellhofer densitometer. It is concluded that MD-55 film, with some limitations, is a useful detector for dosimetry measurements in a single proton beam and can be employed for verification dosimetry in multiple-beam radiation therapy. PMID- 9204524 TI - The use of a multitracer technique for the studies of the uptake and retention of trace elements in rats. AB - The uptake by, and distribution and retention of radioactive isotopes in various organs of Wistar rats were examined using the multitracer technique. A hydrocholoric acid solution (pH 3) of a carrier-free radioactive multitracer was prepared from gold foil irradiated with a 14N beam of 135 MeV nucleon. The solution was administered orally to 12 7-week-old male rats. Urine and faeces were collected and each group of three rats was killed at 1, 2, 3 or 6 days after administration. The percentage of administered dose of the 17 elements, Mn, Co, Zn, As, Rb, Sr, Y, Eu, Gd, Er, Tm, Yb, Lu, W, Re, Ir and Pt in the organs blood and excreta were determined using gamma-ray spectrometry. Each element revealed its characteristic distribution among the different organs, including the blood. These results are discussed and compared with those of single-tracer experiments, and the advantages of the multitracer technique are presented. PMID- 9204525 TI - Decay characteristics of the induced radioactivity in the target cave of a medical cyclotron. AB - After the transfer of the thick copper target plate irradiated with 30 MeV protons from a medical cyclotron, the gamma dose rate in the target cave was monitored every minute for 25 hours using wall-mounted gamma area monitors. The dose rate decay curve was fitted with four exponential functions. By analysing the slope of the exponentials the traces of radioactive 28Al, 56Mn 24Na and 59Fe were identified. The results were used to minimise personnel radiation exposure during maintenance work on the cyclotron. PMID- 9204526 TI - Synthesis of [O-methyl-11C]fluvoxamine--a potential serotonin uptake site radioligand. AB - 5-Methoxy-1-[4-(trifluoromethyl)-phenyl]-1-pentanone-O-(2-aminoethyl)oxi me (fluvoxamine), a potent clinically used antidepressant, was labelled with carbon 11 (t1/2 = 20.4 min) as a potential radioligand for the non-invasive assessment of serotonin uptake sites in the human brain with positron emission tomography (PET). The two-step radiochemical synthesis consisted of O-methylation of an amino-protected desmethyl precursor with [11C]methyl iodide under mild conditions in the presence of tetrabutylammonium hydroxide in acetonitrile, followed by deprotection with trifluoroacetic acid. 5-[11C]Methoxy-1-[4-(trifluoromethyl) phenyl]-1-pentanone-O-(2-aminoethy l) oxime was obtained in > 98% radiochemical purity in 40 min with a radiochemical yield of 4 +/- 2% (non-decay corrected) and a specific radioactivity of 1 +/- 0.5 Ci/mumol. 5-Hydroxy-1-[4-(trifluoromethyl) phenyl]-1-pentanone-O-[2- (tert-butoxycarbonylamino)ethyl]oxime, the precursor for the radiosynthesis of [11C]fluvoxamine, was prepared by a convenient three step synthesis from the pharmaceutical form of fluvoxamine maleate by converting it into the free base, demethylation by trimethyliodosilane and introduction of the BOC-protective group with di-tert-butyl dicarbonate. PMID- 9204527 TI - Synthesis and iodine-125 labelling of glucuronide compounds for combined chemo- and radiotherapy of cancer. AB - Some types of cancer cells have high levels of beta-glucuronidase activity. This enzyme is able to deglucuronidate a variety of glucuronide derivatives on the cell membrane. Either O- or N-glucuronides can be selectively incorporated into the cancer cells. If the aglycone is cytotoxic, the glucuronide can potentially be used as a selective anti-cancer drug in cancers with high levels of beta glucuronidase activity. Nevertheless, in vitro studies carried out by various investigators have shown that the cytotoxicities of several glucuronides in cancer cells are not sufficiently high for their use as effective anti-cancer drugs. For this reason, we have synthesized glucuronide compounds radiolabelled with iodine-125 combining the radiotoxicity of this Auger electron emitter with the chemotoxicity of the aglycone portion of the glucuronide. PMID- 9204529 TI - Uranium contents and associated effective doses in drinking water from Biscay (Spain). AB - The determination of 234U and 238U content in drinking waters treated at four treatment plants supplying water to a set of municipalities located in northern Spain has given mean values of 1.14 mBq/L for 234U and 0.8 mBq/L for 238U. These contents, taking into account the population supplied with water and its distribution in age intervals, have allowed the determination of the annual intake of both radionuclides as well as the mean committed dose due to the ingestion of these radionuclides for which a value of 0.081 microSv/person is obtained. PMID- 9204530 TI - A survey of radon properties in underground shopping centers in Hong Kong. AB - A number of radon-related properties have been surveyed in underground shopping centers in Hong Kong. These parameters include the radon concentration, the total potential alpha energy concentration of radon progeny, the equilibrium factor and the unattached fraction of radon progeny. The mean values recorded for these were 29.2 +/- 7.8 Bq/m3, 3.60 +/- 1.53 mWL, 0.46 +/- 0.16 and 0.05 +/- 0.03, respectively. Based on these figures, we have calculated the average radon dose received by employees in an underground shopping center in Hong Kong to be 0.22 mSv/yr, which represents an approximate increase of 8% over the total dose of about 2.7 mSv/yr received by the average person in Hong Kong. PMID- 9204531 TI - Regulation of ion transport by P2u purinoceptors and alpha 2A adrenoceptors in HT29 cells. AB - X-ray microanalysis was applied to investigate ion transport mediated by P2U purinoceptors and alpha 2A adrenoceptors as well as their interaction in the regulation of the intracellular elemental concentration in HT29 cells. In response to ATP, HT29 cells showed a decrease of intracellular Cl, Na and an increase in Ca. A similar result was observed with UTP, but UTP appeared to be more potent than ATP. On the other hand, UK14,304, an alpha 2 receptor agonist, was found to be capable of reversing the action of both UTP and ATP on ion secretion, and caused a clear increase in intracellular Na and Cl. Moreover, treatment of cells with UK14,304 before exposure to UTP did not induce a decrease in Cl and Na, suggesting that UK14,304 blocks the action of UTP. The secretory effect of UTP was also blocked by NPPB, a chloride channel blocker, and alloxan. Chelation of extracellular Ca with EGTA abolished ion response to UTP. These results suggest that since inhibition of the intracellular cAMP system and chelation of Ca2+ can block the nucleotide-induced chloride secretion, the ATP and UTP-induced chloride secretion can be mediated via both cAMP-dependent and Ca(2+)-dependent pathways. PMID- 9204533 TI - A rapid bioassay to detect mycotoxins using a melanin precursor overproducer mutant of the ciliate Tetrahymena thermophila. AB - Four different mycotoxins (patulin, T-2 toxin, diacetoxyscirpenol and roquefortine) were used to study growth inhibitory effects on a melanin precursor overproducer mutant of the ciliate Tetrahymena thermophila. This strain is especially sensitive to diacetoxyscirpenol and T-2 toxin. The secretion capacity of melanin precursors into the culture medium by this mutant and its biosensor capacity are very useful characteristics to elaborate a rapid bioassay to detect some specific mycotoxins. PMID- 9204532 TI - Characterization of the Na+, K(+)-ATPase in isolated bovine articular chondrocytes; molecular evidence for multiple alpha and beta isoforms. AB - We have used isoform-specific antibodies against the Na+, K(+)-ATPase alpha (alpha 1, alpha 2 and alpha 3) and beta (beta 1 and beta 2) subunit isoforms in order to establish their specific localization in isolated bovine articular chondrocytes. Immunoblotting confirmed the presence of the alpha 1 and alpha 3 isoforms, although alpha 1 expression was significantly greater than alpha 3 as assessed by immunofluorescence confocal laser scanning microscopy and PCR. A similar approach revealed the presence of the beta 1 and beta 2 isoforms in chondrocytes, although beta 2 immunostaining on the plasma membrane was more punctate than beta 1 which in contrast predominated in a subcellular compartment. The plasma membrane abundance of the Na+, K(+)-ATPase was found to be sensitive to the extracellular ionic concentration and long-term elevation of extracellular Na+ concentration significantly upregulated Na+, K(+)-ATPase density as measured by specific 3H-ouabain binding. Our observations suggest that the expression of alpha 3 and beta 2 is not restricted to excitable tissues as previously reported. The physiological relevance of alpha 3 expression in chondrocytes may be related to its low affinity for intracellular Na+ in an extracellular environment where Na+ concentration is unusually high (260-350 mM) compared to other cell types (140 mM). Glycoproteins and their branched carbohydrates have been implicated in cell recognition events, thus the beta 2 subunit glycoprotein may allow the chondrocyte to detect changes in its extracellular environment by physically interacting with components of the cellular cytoskeleton and matrix macromolecules. PMID- 9204534 TI - Choline base exchange activity in rat hepatocyte nuclei and nuclear membranes. AB - Previous investigations have demonstrated the presence of phospholipids as a component of chromatin; however the mechanism of their synthesis, namely if they are synthesized in the nuclei or in the cytoplasm (microsomal fraction), from where they may eventually be transported to the nucleus, has not yet been clarified. The phosphatidylcholine, for example, can be formed, albeit in a limited amount, by an interconversion reaction between bases. The aim of the present research was to ascertain the presence of the enzyme complex responsible for this reaction in hepatocyte nuclei and in isolated nuclear membrane. The incorporation of [14C]-choline in phosphatidylcholine was assayed in microsomes, hepatocyte nuclei, liver nuclei and nuclear membranes of rat liver. The reaction was Ca(2+)-dependent and the specific activity was higher in microsomes but was present, albeit at a low level, also in nuclei and in nuclear membranes. Possible contaminations were excluded by specific microsomal markers and by the reaction time course. In fact, the nuclear reaction reached the maximum level slowly with respect to microsomes. Since the phosphatidylcholine extracted from the nuclei show an enrichment in unsaturated fatty acids of monoenoic fraction, such as oleic acid, the difference in reaction kinetics has been tentatively explained as due to the phosphatidylcholine fatty acid content. The presence of this base exchange enzyme complex may allow a fast change in chromatin phospholipid composition. PMID- 9204535 TI - Expression of a cytoplasmically epitope-tagged human Golgi glycosyltransferase in homologous cells results in mislocalization of multiple Golgi proteins. AB - We have compared the effect of mislocalization of a Golgi glycosyltransferase in heterologous and homologous cell systems on the distribution of other Golgi associated proteins. Mycspacer-human N-acetylglucosaminyltransferase I (NAGT-I), an N-terminally epitope-tagged NAGT-I, in which the first added negatively charged amino acid is in position 13, localizes to the endoplasmic reticulum (ER) by immunofluorescence when expressed in monkey (Vero) or human (HeLa) cells. When myc-spacer-human NAGT-I was expressed in Vero cells, the distribution of the Golgi-associated coat protein, beta-COP, was concentrated juxtanuclearly and undisturbed relative to control. When myc-spacer-human NAGT-I was expressed in HeLa cells, however, both endogenous beta-COP and GalT were no longer concentrated in a juxtanuclear manner but were rather cytoplasmically distributed as was the myc-tagged human NAGT-I. Based on these observation, we suggest that extensive interactions between proteins that normally show overlapping distributions between the medial Golgi stack and trans Golgi/TGN are possible. Moreover, we suggest that small differences in sequence may play a large role in potentiating interactions of Golgi complex proteins. PMID- 9204536 TI - Kinesin-associated transport is involved in the regulation of cell adhesion. AB - It has been recently shown that deploymerization of microtubules induces the elongation of focal contacts at the leading edge. On the other hand, cell shape and pseudopodial activity were found to depend on the microtubule-based motor kinesin. In this paper, we examine whether kinesin is involved in controlling the dynamics of adhesive structures at the cell surface. Microinjection of an antiblocking kinesin activity in vitro causes focal contact elongation similar to the effect of microtubule-depolymerizing drugs. Thus, the role of microtubules in cell adhesion lies in the supporting kinesin-based transport to the adhesion sites. PMID- 9204537 TI - Cisplatin inhibits pinocytosis in Dictyostelium discoideum. AB - The effects of cis-diamminedichloroplatinum(II) [cisplatin], a potential anticancer drug, were studied on pinocytotic functions in the cellular slime mould Dictyostelium discoideum by administering FITC-dextran as a fluid phase marker. Cisplatin treatment at a concentration of 100 and 200 micrograms/ml for 1 h causes inhibition in pinocytotic uptake in growing Dictyostelium cells in a dose-dependent manner. Cisplatin treatment induced the association of more actin with the cell cortex, thereby presumably restricting the flexibility of the cell membrane and inhibiting the formation of pinosomes. Ultrastructural analysis of cisplatin-treated cells showed a lower number of pinosomes. These results have been discussed in the light of cisplatin's known actions that affect various cellular functions. PMID- 9204538 TI - Evidence for a localization signal in the 3'untranslated region of myosin heavy chain messenger RNA. AB - Localization signals in the 3'untranslated region (3'UTR) of myosin heavy chain mRNA were investigated using hybrid gene constructs. In myoblasts transfected with constructs containing either both coding sequences and 3'UTR of the rabbit beta-globin gene or the beta-globin coding sequences alone in situ hybridization showed that globin transcripts were distributed throughout the cytoplasm with no localization. In contrast, in myoblasts transfected with beta-globin coding sequences linked to the myosin heavy chain 3'UTR there was strong perinuclear localization of the hybrid mRNA; this was maintained in myotubes. We conclude that myosin heavy chain 3'UTR contains a localization signal. PMID- 9204539 TI - Opossum kidney cells take up L-DOPA through an organic cation potential-dependent and proton-independent transporter. AB - The present work was aimed at studying the kinetics and nature of the L-DOPA transporter in opossum kidney (OK) cells. Saturation experiments were performed in OK cells incubated for 6 min with increasing concentrations of L-DOPA (10 to 2500 microM); non-linear analysis of the saturation curve revealed for L-DOPA a K(m) of 129 microM (114, 145) and Vmax of 30.0 +/- 0.4 nmol mg protein-1 6 min-1. The uptake of L-DOPA (250 microM) was inhibited in a concentration-dependent manner by cyanine 863, an organic cation inhibitor, with a Ki value of 638 (430, 947) microM; the organic anion inhibitor 4,4'-diisothiocyanostilbene-2,2' disulphonic acid (DIDS), was devoid of effect upon the uptake of L-DOPA. The uptake of L-DOPA (250 microM) was significantly (P < 0.02) decreased (25% reduction) when cells were incubated in the presence of 137 mM K+ plus 5 mM Na+, when compared with the control condition (137 mM Na+ plus 5 mM K+); substitution of NaCl by choline chloride (137 mM) did not affect L-DOPA uptake. Similarly, inwardly or outwardly directed proton gradients of 0.5 pH units (7.9, 7.4, 6.9, 6.4 and 5.9) were found not to change L-DOPA uptake. In conclusion, the L-DOPA uptake system in OK cells has the characteristics of an organic cation potential dependent and proton-independent transporter. PMID- 9204540 TI - Synthesis and characterization of m- and p-methylbenzyl-mercapturic acids derived from m- and p-xylenes. AB - Chemical synthesis and physical properties of two mercapturic acids suggested as urinary metabolites of m- and p-xylenes are described. These compounds may be used for the identification and quantitative determination by high-performance liquid chromatography of the corresponding mercapturic acids in urine. PMID- 9204541 TI - PCDD, PCDF, and PCB concentrations in human milk from two areas in Finland. AB - Concentrations of 17 toxic 2,3,7,8-chlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/Fs) and of 36 polychlorinated biphenyl congeners (PCBs) were analyzed from 167 randomly sampled human milk samples from Southern (77 samples) and Eastern (90) Finland. The level of PCDD/Fs and PCBs in human milk in Southern Finland was about 25% higher than those from Eastern Finland. The level of PCDD/Fs in human milk in Finland was the same as in Sweden, 30-50% lower than in milk from Central Europe but about 45% higher than values from Norway or Russia. The PCB concentrations in Southern Finland were at the same level as in the Netherlands, and in Eastern Finland at the same level as in Norway. The levels of PCDD/Fs and PCBs decreased with increasing number of children: the third child was exposed to about 70% of the amount of PCDD/Fs and PCBs as compared with the first child, and the eighth to tenth child to about 20%. The congener patterns of PCDD/Fs and PCBs in Finnish human milk were similar to the countries of Central Europe, however, the levels of penta- and heptachlorinated furans were slightly higher than in milk from other countries. PMID- 9204542 TI - Decreased blood level of beta 2-microglobulin in the employees of a factory which produced polychlorinated biphenyls. AB - Under the project of evaluating the health status of the employees of CHEMKO factory (East Slovakia) which produced PCBs between 1955 and 1985, the level of beta 2-microglobulin (beta 2-m) was measured in the serum of 242 subjects from CHEMKO (Group A) and two control groups from much less polluted areas: 1,277 females from a small mountainous village (Group B), 2,179 adults from the area of a large city of Kosice (Group C). The level of thymidine kinase (TK) was measured in the groups A and B only. In addition, age-matched groups of 155 women each from all areas were evaluated. In both the whole group and the age-matched group from CHEMKO the level of beta 2-m was significantly lower (P < 0.001) than that in the respective control groups, while no difference was found in the level of TK. In conclusion, it is suggested that the decrease of beta 2-m in CHEMKO employees might be related to the immunotoxic effects of organochlorines. PMID- 9204543 TI - Biodegradation of sucrose poly fatty acid esters in soils. AB - Sucrose polyesters (SPEs) were applied to soil at rates equivalent to 1062 to 1293 kg per hectare and incubated over periods of 100 to 403 days at 20 +/- 2 degrees C in darkness and at a soil moisture of 40% of the maximum water holding capacity. All applied forms of SPEs were aerobically biodegraded to some degree in both American and German soil. However, the mineralization rates varied considerably and were dependent on both SPE and soil type. For example, sucrose octaoleate underwent slow and limited mineralization in the German soils Speyer and Borstel as well as in the American soil Madera, reaching only 6.9-18.4% mineralisation after over 400 days incubation. The same material in the American soils Hollande, Thermal and Uvalde as well as in the German soil Speicherkoog, reached 35-52% after the same incubation period. Of the SPEs most realistic for use in food products, the more liquid (i.e. with the least saturated fatty acids) underwent the most rapid and extensive mineralization. However, the mineralization rates for these materials were distinctly lower than the corresponding ones for sucrose octaoleate. In all cases the extent of mineralization of the SPEs in soil was significantly lower than that of a control fat (synthetic triglyceride mixture HB307), which typically underwent over 50% mineralization in 60 days. PMID- 9204544 TI - Mental time travel and the evolution of the human mind. AB - This article contains the argument that the human ability to travel mentally in time constitutes a discontinuity between ourselves and other animals. Mental time travel comprises the mental reconstruction of personal events from the past (episodic memory) and the mental construction of possible events in the future. It is not an isolated module, but depends on the sophistication of other cognitive capacities, including self-awareness, meta-representation, mental attribution, understanding the perception-knowledge relationship, and the ability to dissociate imagined mental states from one's present mental state. These capacities are also important aspects of so-called theory of mind, and they appear to mature in children at around age 4. Furthermore, mental time travel is generative, involving the combination and recombination of familiar elements, and in this respect may have been a precursor to language. Current evidence, although indirect or based on anecdote rather than on systematic study, suggests that nonhuman animals, including the great apes, are confined to a "present" that is limited by their current drive states. In contrast, mental time travel by humans is relatively unconstrained and allows a more rapid and flexible adaptation to complex, changing environments than is afforded by instincts or conventional learning. Past and future events loom large in much of human thinking, giving rise to cultural, religious, and scientific concepts about origins, destiny, and time itself. PMID- 9204545 TI - Card-sort performance and syndromes of schizophrenia. AB - The performance of subgroups of schizophrenic patients on the Wisconsin Card Sort Test was investigated. Factor analyses of positive and negative symptom ratings confirmed that ratings described three groups similar to those reported by several other researchers. These symptom groups were labeled psychomotor poverty, cognitive disorganization, and reality distortion. Coaching and incentives resulted in significant improvement in the card-sort performance for patients characterized by symptoms of psychomotor poverty and reality distortion. Patients with symptoms of disorganization, however, had impaired ability to improve card sort performance after coaching and incentives. Results indicate that symptoms of disorganization appear to differentiate learners from nonlearners on the Wisconsin Card Sorting Test and may be correlated with different underlying deficits. PMID- 9204546 TI - Dimensions of marital relationships as perceived by Turkish husbands and wives. AB - In this study, the basic underlying dimensions and interrelationships of Turkish urban marriages were explored. Both husbands and wives from 456 marriages of different types, lengths, and socioeconomic status (SES) groups completed the extensive Turkish Marriage Questionnaire (Russell, Wells, & Imamoglu, 1989). First-order factor analysis yielded 9 factors that were then reduced to 4 second order factors: Extent of Socioeconomic Development, Marital Satisfaction, Harmonious Relations With the Extended Family, and Desire for Sexual Possessiveness. The frequency of self-selected marriages increased with higher SES and decreased with length of marriage, implying a trend toward modernism. Within this context, husbands' marital satisfaction and wives' desire for sexual possessiveness, extent of socioeconomic development, and relations with the extended family were significant predictors of wives' marital satisfaction; husbands' marital satisfaction was predicted by wives' satisfaction and husbands' relations with the extended family. PMID- 9204547 TI - Evidence for a regulatory protein complex on RNA polymerase III promoter of human retroposons of Alu family. AB - Abundant human retroposons of the Alu family produce few RNA polymerase III (RPIII)-dependent transcripts in vivo. This suggests that either the bulk of the repeats has no proper promoter elements or that transcription of Alu by RPIII is repressed. In this study, we analyzed complexes formed by human nuclear proteins with the Alu B-box and with an adjacent downstream sequence (DB-sequence). Four complexes (C1-C4) were detected and two of them (C2 and C3) were found to be induced by different proteins. C3 formation was found to be sensitive to minor sequence variation within the Alu DB-sequence. The C2 complex is specifically repressed by the competing VA1 B-box oligonucleotide and was found to be very stable. In addition, it is downregulated in human cells transformed by adenovirus 5. This is consistent with a view that the C2 complex is formed by a protein (designated as ACR1) that is different from TFIIIC2. The ACR1 protein may be involved in the modulation of Alu transcription in vivo by interfering or cooperating with TFIIIC2. A similar complex is detected with the efficiently transcribed adenovirus VA1 RNA gene B-box. We compared the affinity of complexes formed by ACR1 with Alu and VA1 B-boxes. It was found that both B-boxes bind ACR1 with equal affinity with a dissociation constant of about 2 nM. However, DB sequences in Alu and VA1 promoters are non-homologous, and C3/C4 complexes are found to be formed with Alu DB, but not formed with VA1 DB sequences. The Alu specific protein forming C3 (named as ACR2) may cooperate with ACR1 in selective repression of RPIII-dependent Alu transcription in vivo. PMID- 9204548 TI - Identification of new medium reiteration frequency repeats in the genomes of Primates, Rodentia and Lagomorpha. AB - We report eleven new families of MEdium Reiteration frequency (MER) interspersed repeats in the genomes of Primates, Rodentia, and Lagomorpha. Two families of the human repeats, MER 46 and MER 47, represent non-autonomous DNA transposons. These sequences are flanked by TA target site duplications and have terminal inverted repeats (TIRs) similar to TIRs of DNA transposons. The sequences of five other families of repeats, MER41, MER48, MER50, MER51, and RMER3, resemble long terminal repeats of retroviruses. A potential involvement of some of the reported MER repeats in the regulation of transcription and genetic rearrangements is suggested. Age estimations place the origin of most MER repeats at the time of decline in MIR (Mammalian-wide Interspersed Repeats) retroposition and before the origin of the Alu family. PMID- 9204550 TI - Cloning and characterization of a 70 kd heat shock cognate (hsc70) gene from the zebrafish (Danio rerio). AB - The heat shock 70 family of proteins is one of the most highly conserved among all species. The genes encoding these proteins have been cloned and sequenced from bacterial species to humans with a high degree of homology preserved throughout evolution. Here we describe the cloning and characterization of a cDNA encoding a 70 kd heat shock cognate (hsc70) gene from the zebrafish (Danio rerio). A high degree of conservation is observed among hsc70 genes of other species as shown by phylogenetic analysis. The characterization of a hsc70 gene in the zebrafish provides a marker for studying the role of a constitutively expressed member of the hsp70 family in an important developmental and evolutionary model system. PMID- 9204549 TI - Molecular characterization of the cinnabar region of Drosophila melanogaster: identification of the cinnabar transcription unit. AB - Early studies of eye pigmentation in Drosophila melanogaster provided compelling evidence that the cinnabar (cn) gene encodes the enzyme kynurenine 3 monooxygenase (EC 1.14.13.9). Here we report the cloning of approximately 60 kb of DNA encompassing the cn gene by chromosome walking in the 43E6-F1 region of chromosome 2. An indication of the position of cn within the cloned region was obtained by molecular analysis of mutants: 9 spontaneous cn mutants were found to have either DNA insertions or deletions within a 5 kb region. In addition, a 7.8 kb restriction fragment encompassing the region altered in the mutants was observed to induce transient cn function when microinjected into cn- embryos. The cn transcription unit was identified by Northern blotting and sequence analysis of cDNA and genomic clones from this region. The predicted cn protein contains several sequence motifs common to aromatic monooxygenases and is consistent with the assignment of cn as encoding the structural gene for kynurenine 3 monooxygenase. PMID- 9204551 TI - Human phylogenetic relationships according to the D1S80 locus. AB - By analyzing the allelic frequencies at the D1S80 locus in 43 human populations, we show that the locus is polymorphic globally and that it can be used to discriminate between major racial groups and subpopulations through phylogenetic analysis. Although the use of informative multiple loci generally provides more accurate phylogenetic relationships, in instances where time and/or target DNA availability is limited, D1S80 could provide useful data to discriminate between human groups. Also, knowledge of which loci independently provide accurate phylogenetic relationships, such as the D1S80, can be used to design more accurate multi-locus combinations. In addition, allele frequencies at the locus are reported, for the first time, for Bahamian individuals of African origin and for Chimila, Bari, and Navajo (Canoncito Valley) native Americans. Allelic data was obtained using standard polymerase chain reaction (PCR) techniques. In the four new populations, 65 genotypes and 20 segregating alleles were observed. All populations conformed to Hardy-Weinberg expectations except the Chimila. PMID- 9204552 TI - Analysis of inbreeding in a colonizing population of Drosophila subobscura. AB - An analysis of the effects of inbreeding on the genetic structure of a colonizing population of Drosophila subobscura has been carried out. Species of Drosophila, particularly D. subobscura, may have lethal alleles associated with chromosomal inversions and our aim was to assess the extent to which the genome is balanced in this way. The frequencies of chromosomal inversions were compared between a large population and a set of 72 lines that were maintained by brother-sister mating for 10 generations. Fisher's matrix method was used to calculate the expected homozygosity in these inbred lines for 5 allozyme loci (Aph, Hk-1, Lap, Odh, and Pept-1) used as markers of large chromosomal segments. Furthermore, the expected rates of fixation corresponding to these allozyme loci were also calculated. The results show that the amount of homozygosis observed did not differ significantly from expectations (with the corresponding loss of lines as a consequence of the reduction in viability). However, two deviations from strict neutrality were observed: there was a heterozygote excess at the Lap locus, and the frequency of the O5 inversion (always associated with a lethal gene in colonizing populations) was higher than expected. PMID- 9204553 TI - Solution structure of human intestinal fatty acid binding protein: implications for ligand entry and exit. AB - The human intestinal fatty acid binding protein (I-FABP) is a small (131 amino acids) protein which binds dietary long-chain fatty acids in the cytosol of enterocytes. Recently, an alanine to threonine substitution at position 54 in I FABP has been identified which affects fatty acid binding and transport, and is associated with the development of insulin resistance in several populations including Mexican-Americans and Pima Indians. To investigate the molecular basis of the binding properties of I-FABP, the 3D solution structure of the more common form of human I-FABP (Ala54) was studied by multidimensional NMR spectroscopy. Recombinant I-FABP was expressed from E. coli in the presence and absence of 15N enriched media. The sequential assignments for non-delipidated I-FABP were completed by using 2D homonuclear spectra (COSY, TOCSY and NOESY) and 3D heteronuclear spectra (NOESY-HMQC and TOCSY-HMQC). The tertiary structure of human I-FABP was calculated by using the distance geometry program DIANA based on 2519 distance constraints obtained from the NMR data. Subsequent energy minimization was carried out by using the program SYBYL in the presence of distance constraints. The conformation of human I-FABP consists of 10 antiparallel beta-strands which form two nearly orthogonal beta-sheets of five strands each, and two short alpha-helices that connect the beta-strands A and B. The interior of the protein consists of a water-filled cavity between the two beta-sheets. The NMR solution structure of human I-FABP is similar to the crystal structure of rat I-FABP. The NMR results show significant conformational variability of certain backbone segments around the postulated portal region for the entry and exit of fatty acid ligand. PMID- 9204554 TI - The dynamic NMR structure of the T psi C-loop: implications for the specificity of tRNA methylation. AB - tRNA (m5U54)-methyltransferase (RUMT) catalyzes the S-adenosylmethionine dependent methylation of uridine-54 in the T psi C-loop of all transfer RNAs in E. coli to form the 54-ribosylthymine residue. However, in all tRNA structures, residue 54 is completely buried and the question arises as to how RUMT gains access to the methylation site. A 17-mer RNA hairpin consisting of nucleotides 49 65 of the T psi-loop is a substrate for RUMT. Homonuclear NMR methods in conjunction with restrained molecular dynamics (MD) methods were used to determine the solution structure of the 17-mer T-arm fragment. The loop of the hairpin exhibits enhanced flexibility which renders the conventional NMR average structure less useful compared to the more commonly found situation where a molecule exists in predominantly one major conformation. However, when resorting to softer refinement methods such as MD with time-averaged restraints, the conflicting restraints in the loop can be satisfied much better. The dynamic structure of the T-arm is represented as an ensemble of 10 time-clusters. In all of these, U54 is completely exposed. The flexibility of the T psi-loop in solution in conjunction with extensive binding studies of RUMT with the T psi C loop and tRNA suggest that the specificity of the RUMT/ tRNA recognition is associated with tRNA tertiary structure elements. For the methylation, RUMT would simply have to break the tertiary interactions between the D- and T-loops, leading to a melting of the T-arm structure and making U54 available for methylation. PMID- 9204555 TI - Assignment methodology for larger RNA oligonucleotides: application to an ATP binding RNA aptamer. AB - The use of uniform 13C, 15N labeling in the NMR spectroscopic study of RNA structures has greatly facilitated the assignment process in small RNA oligonucleotides. For ribose spin system assignments, exploitation of these labels has followed previously developed methods for the study of proteins. However, for sequential assignment of the exchangeable and nonexchangeable protons of the nucleotides, it has been necessary to develop a variety of new NMR experiments. Even these are of limited utility in the unambiguous assignment of larger RNAs due to the short carbon relaxation times and extensive spectral overlap for all nuclei. These problems can largely be overcome by the additional use of basetype selectively 13C, 15N-labeled RNA in combination with a judicious use of related RNAs with base substitutions. We report the application of this approach to a 36-nucleotide ATP-binding RNA aptamer in complex with AMP. Complete sequential 1H assignments, as well as the majority of 13C and 15N assignments, were obtained. PMID- 9204557 TI - Rotational diffusion anisotropy of proteins from simultaneous analysis of 15N and 13C alpha nuclear spin relaxation. AB - Current methods of determining the rotational diffusion tensors of proteins in solution by NMR spectroscopy exclusively utilize relaxation rate constants for backbone amide 15N spins. However, the distributions of orientations of N-H bond vectors are not isotropic in many proteins, and correlations between bond vector orientations reduce the accuracy and precision of rotational diffusion tensors extracted from 15N spin relaxation data. The inclusion of both 13C alpha and 15N spin relaxation rate constants increases the robustness of the diffusion tensor analysis because the orientations of the C alpha-H alpha bond vectors differ from the orientations of the N-H bond vectors. Theoretical and experimental results for calbindin D9k, granulocyte colony stimulating factor, and ubiquitin, three proteins with different distributions of N-H and C alpha-H alpha bond vectors, are used to illustrate the advantages of the simultaneous utilization of 13C alpha and 15N relaxation data. PMID- 9204556 TI - NMR secondary structure of the plasminogen activator protein staphylokinase. AB - Staphylokinase (Sak) is a 15.5 kDa protein secreted by several strains of Staphylococcus aureus. Due to its ability to convert plasminogen, the inactive proenzyme of the fibrinolytic system, into plasmin, Sak is presently undergoing clinical trials for blood clot lysis in the treatment of thrombovascular disorders. With a view to developing a better understanding of the mode of action of Sak, we have initiated a structural investigation of Sak via multidimensional heteronuclear NMR spectroscopy employing uniformly 15N- and 15N, 13C-labelled Sak. Sequence-specific resonance assignments have been made employing 15N-edited TOCSY and NOE experiments and from HNCACB, CBCA(CO)NH, HBHA-(CBCACO) NH and CC(CO)NH sets of experiments. From an analysis of the chemical shifts, 3JHNH alpha scalar coupling constants, NOEs and HN exchange data, the secondary structural elements of Sak have been characterized. PMID- 9204558 TI - Chi 1 angle information from a simple two-dimensional NMR experiment that identifies trans 3JNC gamma couplings in isotopically enriched proteins. AB - New quantitative J correlation experiments are used for measuring all two- and three-bond couplings between 15N and aliphatic side-chain carbons in proteins uniformly enriched in 13C and 15N. Results show that 3JNC beta and 2JNC beta invariably are very small. Therefore, a simple and relatively sensitive two dimensional spin-echo difference experiment can be used to identify residues with a 3JN gamma coupling substantially larger than 1 Hz, indicative of a trans arrangement between N and C gamma. This measurement therefore provides chi 1 angle information for residues with an aliphatic C gamma carbon, and thereby also aids in making stereospecific assignments of H beta resonances. Experiments are demonstrated for ubiquitin and for a complex between calmodulin and a 26-residue peptide. PMID- 9204559 TI - Salicylate-independent lesion formation in Arabidopsis lsd mutants. AB - In many interactions of plants with pathogens, the primary host defense reaction is accompanied by plant cell death at the site of infection. The resulting lesions are correlated with the establishment of an inducible resistance in plants called systemic acquired resistance (SAR), for which salicylic acid (SA) accumulation is a critical signaling event in Arabidopsis and tobacco. In Arabidopsis, the lesions simulating disease (lsd) mutants spontaneously develop lesions in the absence of pathogen infection. Furthermore, lsd mutants express SAR marker genes when lesions are present and are resistant to the same spectrum of pathogens as plants activated for SAR by necrogenic pathogen infection. To assess the epistatic relationship between SA accumulation and cell death, transgenic Arabidopsis unable to accumulate SA due to the expression of the salicylate hydroxylase (nahG) gene were used in crosses with the dominant mutants lsd2 or lsd4. Progeny from the crosses were inhibited for SAR gene expression and disease resistance. However, these progeny retained the spontaneous cell death phenotype similar to siblings not expressing nahG. Because lesions form in the absence of SA accumulation for isd2 and lsd4, a model is suggested in which lesion formation in these two mutants is determined prior to SA accumulation in SAR signal transduction. By contrast, the loss of SAR gene expression and disease resistance in nahG-expressing lsd mutants indicates that these traits are dependent upon SA accumulation in the SAR signal transduction pathway. PMID- 9204560 TI - Synthetic peptide combinatorial libraries: a method for the identification of bioactive peptides against phytopathogenic fungi. AB - Synthetic combinatorial libraries were evaluated with an iterative process to identify a hexapeptide with broadspectrum activity against selected phytopathogenic fungi. A D-amino acid hexapeptide (FRLKFH) and pentapeptide (FRLHF) exhibited activity against Fusarium oxysporum f. sp. lycopersici, Rhizoctonia solani (anastomosis group 1), Ceratocystis fagacearum, and Pythium ultimum. The peptides showed no hemolytic or mutagenic activity. Fluorescent microscopy studies with a membrane impermeant dye indicated that fungal cytoplasmic membranes were compromised rapidly and that the nuclear membrane was also affected. PMID- 9204562 TI - Specific flavonoids promote intercellular root colonization of Arabidopsis thaliana by Azorhizobium caulinodans ORS571. AB - The ability of Azorhizobium caulinodans ORS571 and other diazotrophic bacteria to internally colonize roots of Arabidopsis thaliana has been studied. Strains tagged with lacZ or gusA reporter genes were used, and the principal colonization sites were found to be the points of emergence of lateral roots, lateral root cracks (LRCs). High frequencies of colonization were found; 63 to 100% of plants were colonized by ORS571, and 100% of plants were colonized by Herbaspirillum seropedicae. After LRCs were colonized, bacteria moved into intercellular spaces between the cortical and endodermal cell layers. Specific flavonoids, naringenin and daidzein, at 5 x 10(-5) M, significantly promoted colonization by ORS571. By using a nodC and a nodD mutant of ORS571, it was shown that neither Nod factors nor NodD are involved in colonization or flavonoid stimulation of colonization. Flavonoids did not appear to be stimulating LRC colonization by their activity as nutritional factors. LRC and intercellular colonization by H. seropedicae was stimulated by naringenin and daidzein at the same concentration that stimulated colonization by ORS571. PMID- 9204561 TI - Identification and characterization of a gene on Rhizobium meliloti pSyma, syrB, that negatively affects syrM expression. AB - The Rhizobium meliloti SyrM protein activates transcription of nodD3 and syrA. Regulation of syrM is complex and may involve as yet undiscovered genes. Here we report the isolation of insertion mutants showing increased expression of a syrM gusA gene fusion. Characterization of one mutant strain, designated SYR-B, revealed a mutation consisting of a transposon insertion linked to a large deletion. The corresponding wild-type DNA was cloned as a 5.3-kb BamHI fragment. Genetic and physical analysis of this DNA demonstrated that an open reading frame (ORF) near one end of the fragment, encoding the 16.5-kDa SyrB protein, is responsible for the repression of syrM activity. Results of complementation experiments with the 5.3-kb BamHI DNA led us to hypothesize that other genes within this DNA fragment interfere with the expression or activity of SyrB. Our analysis showed that the region upstream of syrB contains three ORFs. One ORF is similar to the Ros repressor of Agrobacterium tumefaciens and the MucR repressor of R. meliloti. PMID- 9204566 TI - Functional and regulatory analysis of the two copies of the fixNOQP operon of Rhizobium leguminosarum strain VF39. AB - DNA corresponding to two copies of the Rhizobium leguminosarum bv. viciae strain VF39 fixNOQP operon coding for a putative symbiotic terminal oxidase of the heme copper oxidase superfamily was cloned, sequenced, and genetically analyzed. The first copy is located upstream of the fixK-fixL region on plasmid pRleVF39c, whereas the second copy resides on the nodulation plasmid pRleVF39d. Insertional mutagenesis with antibiotic resistance cassettes confirmed that both copies were functional, and that the presence of at least one functional copy was required for nitrogen fixation. The deduced amino acid sequences of both fixN genes are highly similar (95% identity) and contain 15 putative transmembrane helices, suggesting that the fixN gene products are integral membrane proteins. Furthermore, six histidine residues predicted to be the ligands for a heme-copper binuclear center and a low-spin heme b are conserved in both R. leguminosarum fixN proteins. The deduced fixO and fixP gene products show characteristics of membrane-bound monoheme and diheme cytochrome c, respectively. Upstream of both fixN copies putative Fnr-consensus binding sites (anaeroboxes) were found that differ in certain base pairs. As R. leguminosarum VF39 possesses two members of the Fnr/FixK regulator family, FnrN and FixK, the possible differential regulation of both fixN copies was analyzed with fixN-gusA reporter gene fusions. Both fixN fusions were induced under free-living microaerobic conditions and in the symbiotic zone of the root nodule. Induction of the expression of fixNc and fixNd was highly reduced in a fnrN mutant background and in a fixL mutant background, whereas fixK was only marginally involved in fixN regulation. Residual expression of fixN was observed in an fnrN/fixK double mutant. PMID- 9204563 TI - Evidence that the Pseudomonas syringae pv. syringae hrp-linked hrmA gene encodes an Avr-like protein that acts in an hrp-dependent manner within tobacco cells. AB - A 25-kb DNA region, previously cloned from Pseudomonas syringae pv. syringae 61 in cosmid pHIR11, enables nonpathogenic bacteria such as Pseudomonas fluorescens and Escherichia coli to elicit the hypersensitive response (HR) in tobacco (Nicotiana tabacum). hrmA is located within this region, adjacent to a conserved cluster of hrp genes, and is essential for nonpathogens to elicit the HR. DNA sequence analysis suggested that hrmA was the second of two genes in an operon and was preceded by an open reading frame (ORF), ORF1, which is predicted to encode a 10.9-kDa protein. DNA gel blot analysis revealed that sequences hybridizing with a DNA fragment internal to hrmA were absent from P. syringae pv. syringae B728a, P. syringae pv. tabaci 11528, and P. syringae pv. glycinea race 4 U1, but present in P. syringae pv. tomato DC3000. A 2.4-kb BamHI-AvrII fragment carrying hrmA, ORF1, and native regulatory sequences was subcloned into broad host-range vector pDSK519 and electroporated into P. syringae pv. syringae B728a and P. syringae pv. tabaci 11528. The presence of the hrmA locus had no apparent effect on the ability of P. syringae pv. syringae B728a to cause brown spot of bean, but it caused P. syringae pv. tabaci 11528 to elicit the defense-associated HR rather than disease in N. tabacum cvs. Xanthi N and Xanthi NC and N. clevelandii. Furthermore, N. debeyii, N. glutinosa, N. rustica, and N. tabacum cvs. Petit Havana and Samsun responded with the HR to P. fluorescens(pHIR11). In contrast, N. benthamiana-P. syringae pv. tabaci interactions were unaffected by the presence of HrmA, and P. fluorescens(pHIR11) did not elicit the HR in N. benthamiana. The hrmA ORF was subcloned into pFLAG-CTC, which expressed HrmA with a C-terminal FLAG synthetic epitope fusion. Escherichia coli MC4100 cells carrying the functional hrp cluster and the hrmA-FLAG derivative secreted the HrpZ harpin, but not HrmA-FLAG, to the medium, as indicated by immunoblot analysis with M2 anti-FLAG and polyclonal anti-HrpZ antibodies. The hrmA ORF was also subcloned into plant expression vector pFF19 and then biolistically delivered, along with pFF19G (expressing beta-glucuronidase), into suspension cultured tobacco cells. Histochemical staining 24 h later revealed substantial beta-glucuronidase activity in cells receiving pFF19G and pFF19 but not in those receiving pFF19G and pFF19-HrmA. Thus, internal production of HrmA was deleterious to tobacco cells. PMID- 9204567 TI - Two PR-1 genes from tomato are differentially regulated and reveal a novel mode of expression for a pathogenesis-related gene during the hypersensitive response and development. AB - Pathogenesis-related (PR) proteins form a heterogeneous family of plant proteins that are likely to be involved in defense and are inducible by pathogen attacks. One group of PRs, represented by the subfamily PR-1, are low-molecular-weight proteins of unknown biochemical function. Here we describe the cloning and characterization of two closely related genes encoding a basic and an acidic PR-1 protein (PR1b1 and PR1a2) from tomato (Lycopersicon esculentum). We present a comparative study of the mode of transcriptional regulation of these two genes in transgenic tobacco plants using a series of promoter-GUS fusions. Unexpectedly, the chimeric PR1a2/GUS gene is not induced by pathogenic signals but instead shows constitutive expression with a reproducible developmental expression pattern. It is expressed in shoot meristems, trichomes, and cortical cells as well as in vascular and nearby tissues of the mature stem. This constitutive expression pattern may represent preemption of plant defenses against potential pathogens. Conversely, the chimeric PR1b1/GUS gene does not show any constitutive expression in the plant, but it is transcriptionally activated following pathogen attack. Upon infection by tobacco mosaic virus, the PR1b1 gene is strongly activated locally in tissues undergoing the hypersensitive response but not systemically in uninoculated tissues. Furthermore, its expression is induced by both salicylic acid and ethylene precursors, two signals that coexist and apparently mediate the activation of local defenses during the hypersensitive response. We speculate that the different mode of expression of the two genes presented here, together with that reported previously for the induction of other PR-1 genes in systemic, uninoculated tissues, may all be complementary and necessary for the plant to acquire an efficient refractory state to resist pathogen attacks. PMID- 9204568 TI - Characterization of acquired resistance in lesion-mimic transgenic potato expressing bacterio-opsin. AB - The lesion-mimic mutants of certain plants display necrotic lesions resembling those of the hypersensitive response and activate local and systemic defense responses in the absence of pathogens. We have engineered a lesion-mimic phenotype in transgenic Russet Burbank potato plants through constitutive expression of a bacterio-opsin (bO) proton pump derived from Halobacterium halobium. Transgenic potato plants exhibiting a lesion-mimic phenotype had increased levels of salicylic acid and overexpressed several pathogenesis-related messenger RNAs, all hallmarks of systemic acquired resistance (SAR). The lesion mimic plants also displayed enhanced resistance to the US1 isolate (A1 mating type) of a fungal pathogen, Phytophthora infestans, a causal agent of late blight disease. In contrast, little resistance was observed against the US8 isolate (A2 mating type) of this pathogen. Furthermore, a majority of the transgenic plants displaying the lesion-mimic phenotype had increased susceptibility to potato virus X. The tubers of these plants were not resistant to the bacterial pathogen Erwinia carotovora. These results indicate that expression of bO can result in the activation of defense responses in transgenic potato plants and show for the first time that bO expression can confer resistance to a pathogenic fungus. However, our results also demonstrate that like SAR, this "engineered" resistance is likely to be limited to certain pathogens and particular cultivars. PMID- 9204569 TI - A nodule-specific gene family from Alnus glutinosa encodes glycine- and histidine rich proteins expressed in the early stages of actinorhizal nodule development. AB - Two cDNAs representing different members (agNt84 and ag164) of a gene family encoding glycine- and histidine-rich proteins have been isolated from cDNA libraries from Alnus glutinosa root nodules. Expression of the corresponding genes could only be detected in nodules. With in situ hybridization, the expression in nodules was found to occur in young, infected cells of the prefixation zone (zone 2). The encoded proteins contain putative signal peptides for targeting to the endomembrane system, sharing sequence similarity with signal peptides from plant glycine-rich proteins, among them nodulin 24, a nodule specific protein from soybean. This similarity suggests that, analogous to nodulin-24, proteins encoded by agNt84/ag164 may be located at the interface between the host plant membrane and the matrix surrounding the endosymbiont. The 3' untranslated regions of the cDNAs contain unusual poly(AT)n stretches that may play a role in the regulation of RNA stability. The protein encoded by agNt84 cDNA was expressed in Escherichia coli as a fusion with maltose-binding protein, and was shown to have the ability to bind to a nickel-chelating resin, indicating that it may function as a metal-binding protein. PMID- 9204571 TI - The presence of hrp genes on the pathogenicity-associated plasmid of the tumorigenic bacterium Erwinia herbicola pv. gypsophilae. AB - The pathogenicity-associated plasmid (pPATH) of Erwinia herbicola pv. gypsophilae (Ehg), which is present only in pathogenic strains, contains a gene cluster encoding indole-3-acetic acid and cytokinin biosynthesis. The transposon-reporter Tn3-Spice was used to generate nonpathogenic mutants on two overlapping cosmids, pLA150 and pLA352, of the pPATH. A cluster of such mutations, which spanned 16 kb, mapped approximately 15 kb from the gene cluster involved in phytohormone biosynthesis. Non-pathogenic mutants also failed to elicit the hypersensitive reaction (HR) on tobacco. Pathogenicity and HR were restored concomitantly to these mutants by in trans complementation with wild-type Ehg DNA. A 3.8-kb HindIII DNA fragment that complemented the hrp mutants was sequenced and six complete and two partial open reading frames (ORFs) were identified. Comparison of the deduced amino acid sequences of the eight ORFs showed striking homology and co-linearity with hrp genes of E. amylovora as well as with other plant and mammalian pathogenic bacterial genes encoding proteins of the type III secretion system. Limited DNA sequencing at various sites on the remaining 11-kb region of pLA352 also showed high identity to Hrp proteins of E. amylovora, E. stewartii, and Pseudomonas syringae. These results suggest that hrp genes are mandatory for gall formation by E. herbicola pv. gypsophilae. PMID- 9204570 TI - Evolutionary diversity of symbiotically induced nodule MADS box genes: characterization of nmhC5, a member of a novel subfamily. AB - Unique organs called nodules form on legume roots in response to intracellular infection by soil bacteria in the genus Rhizobium. This study describes a new MADS box gene, nmhC5, which along with nmh7 (J. Heard and K. Dunn, Proc. Natl. Acad. Sci. USA 92:5273-5277, 1995), is expressed in alfalfa (Medicago sativa) root nodules. Together, these genes represent the first putative transcription factors identified in nodules. Expression in a root-derived structure supports the growing sentiment that MADS box proteins have diverse roles in plant development. Comparison of the putative translation product of nmhC5 with those of other reported members of the MADS box family suggests that the overall structure of nmhC5 is conserved. Evolutionary analysis among the MADS box family showed that nmhC5 is orthologous to a root-expressed clone in Arabidopsis thaliana, agl17, and that nmh7 is similar to the floral subfamily with AP3 (DefA)/PI (Glo). Consistent with a prediction of homodimer formation, NMHC5 was shown to bind a CArG consensus sequence in vitro. In contrast, NMH7, which shows structural similarity to MADS box proteins that form heterodimers, did not bind the CArG element in an in vitro DNA-binding assay, suggesting the existence of an unknown dimer partner. The root-derived MADS box genes constitute a novel subfamily of vegetatively expressed MADS box genes. The evolutionary diversity between nmh7 and nmhC5 could represent an overall mechanistic conservation between plant developmental processes or could mean that nmh7 and nmhC5 make fundamentally different contributions to the development of the nodule. PMID- 9204572 TI - Nod factors of Azorhizobium caulinodans strain ORS571 can be glycosylated with an arabinosyl group, a fucosyl group, or both. AB - In addition to the previously described arabinosylated Nod factors, Azorhizobium caulinodans can also produce fucosylated Nod factors and Nod factors that are both arabinosylated and fucosylated. The presence of a plasmid carrying extra copies of a subset of nod genes as well as bacterial growth conditions influence the relative proportion of carbamoylated, fucosylated, and arabinosylated Nod factors. By using a root hair formation assay, we demonstrate that the Nod factor glycosylations are important for biological activity on Sesbania rostrata roots. PMID- 9204573 TI - Improved resolution in the detection of oligonucleotides up to 60-mers in matrix assisted laser desorption/ionization time-of-flight mass spectrometry using pulsed-delayed extraction with a simple high voltage transistor switch. AB - Pulsed-delayed extraction using a simple high voltage transistor switch together with various sample purification approaches were used to enhance the resolution in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) of oligonucleotides up to 60 bases long. This switch can provide a 0-3 kV voltage pulse with a 75 ns fall time. A resolution of 500-900 was typically observed for samples from 5-mers to 60-mers using pulsed-delayed extraction (PDE) with the switch described herein. The resolution deteriorated to < 100 for oligonucleotides of > or = 65-mers. With the TOF acceleration region configuration used in this work, the resolution was found not to vary significantly over a delay range of 2-5 mus. As the DNA size increased to over 35 mer, HPLC purification was required to retain the enhancement in resolution provided by PDE MALDI-MS. PMID- 9204574 TI - New caerin antibacterial peptides from the skin glands of the Australian tree frog Litoria xanthomera. Part 2. Sequence determination using mass spectrometry and associated techniques. AB - Mass spectrometric sequencing, enzymic digestion and Edman degradation provide the structures of the two antimicrobial peptides from the skin glands of the Australian tree frog Litoria xanthomera as:- Gly Leu Phe Ser Val Leu Gly Ala Val Ala Lys His Val Leu Pro His Val Val Pro Val Ile Ala Glu Lys Leu (NH2) (caerin 1.6), and Gly Leu Phe Lys Val Leu Gly Ser Val Ala Lys His Leu Leu Pro His Val Ala Pro Val Ile Ala Glu Lys Leu (NH2) (caerin 1.7). PMID- 9204575 TI - Structural heterogeneity, post-translational modifications, and biological activities of SV-IV, a major protein secreted from the rat seminal vesicle epithelium. AB - The primary structure of purified SV-IV, a major secretory protein synthesized by the rat seminal vesicle (SV) epithelium, was analysed by electrospray mass spectrometry (ES-MS). The protein was found to be highly heterogeneous. The various components were separated and identified by reversed phase high performance liquid chromatography (HPLC) on line with ES-MS. Structural characterization of the SV-IV cyanogen bromide digests revealed the occurrence of a Val/Met substitution in about 50% of the purified protein molecules. We suggest that this mutation is the expression of a genetic polymorphism. Other minor components, corresponding to structural changes (fragmentation, deletion, and phosphorylation) of SV-IV and probably due to post-translational modifications of the native protein, were also detected. In particular, by using protein tyrosine phosphatase hydrolysis combined with ES-MS, we demonstrated that, in the phosphorylated species of SV-IV, a single phosphate group was covalently bound to the Tyr-36 residue. The significance of these findings in relation to the regulation of important biological processes, such as immune response, blood coagulation, inflammatory reaction, and mammalian reproduction, are discussed. PMID- 9204576 TI - Rapid 'de novo' peptide sequencing by a combination of nanoelectrospray, isotopic labeling and a quadrupole/time-of-flight mass spectrometer. AB - Protein microanalysis usually involves the sequencing of gel-separated proteins available in very small amounts. While mass spectrometry has become the method of choice for identifying proteins in databases, in almost all laboratories 'de novo' protein sequencing is still performed by Edman degradation. Here we show that a combination of the nanoelectrospray ion source, isotopic end labeling of peptides and a quadrupole/ time-of-flight instrument allows facile read-out of the sequences of tryptic peptides. Isotopic labeling was performed by enzymatic digestion of proteins in 1:1 16O/18O water, eliminating the need for peptide derivatization. A quadrupole/time-of-flight mass spectrometer was constructed from a triple quadrupole and an electrospray time-of-flight instrument. Tandem mass spectra of peptides were obtained with better than 50 ppm mass accuracy and resolution routinely in excess of 5000. Unique and error tolerant identification of yeast proteins as well as the sequencing of a novel protein illustrate the potential of the approach. The high data quality in tandem mass spectra and the additional information provided by the isotopic end labeling of peptides enabled automated interpretation of the spectra via simple software algorithms. The technique demonstrated here removes one of the last obstacles to routine and high throughput protein sequencing by mass spectrometry. PMID- 9204577 TI - Structural characterization of mono- and dihydroxylated metabolites of paclitaxel in rat bile using liquid chromatography/ion spray tandem mass spectrometry. AB - The capability of high performance liquid chromatography/ion spray mass spectrometry (HPLC/ISP-MS) and HPLC/ISP-tandem mass spectrometry (HPLC/ISP-MS/MS) were investigated to achieve mass separation as well as structural characterization of taxol metabolites directly in rat bile, without their previous isolation. HPLC/ISP-MS yielded information on the molecular weights of several hydroxylated derivatives while HPLC/ISP-MS/MS allowed the on-line structural characterization of all metabolites present in different ratios in rat bile. The approach led to the extraction of nine metabolites and their distinction from the other endogenous contaminants. These metabolites were recognized as three dihydroxytaxols, four monohydroxytaxols, one deacetyltaxol and one containing the taxane ring. Among the derivatives, we were able to identify four new metabolites of paclitaxel belonging to the dihydroxy and monohydroxy series, never previously detected. HPLC/ISP-MS/MS enabled the classification of all di- and monohydroxy isomers. These results demonstrate that the high sensitivity of this method, based on the combined use of tandem mass spectrometry with chromatographic separation, can be considered as offering a valid approach to the detection of new taxol derivatives directly in biological fluids. PMID- 9204578 TI - Automated sample preparation using membrane microtiter extraction for bioanalytical mass spectrometry. AB - The development and application of membrane solid phase extraction (SPE) in 96 well microtiter plate format is described for the automated analysis of drugs in biological fluids. The small bed volume of the membrane allows elution of the analyte in a very small solvent volume, permitting direct HPLC injection and negating the need for the time consuming solvent evaporation step. A programmable liquid handling station (Quadra 96) was modified to automate all SPE steps. To avoid drying of the SPE bed and to enhance the analytical precision a novel protocol for performing the condition, load and wash steps in rapid succession was utilized. A block of 96 samples can now be extracted in 10 min., about 30 times faster than manual solvent extraction or single cartridge SPE methods. This processing speed complements the high-throughput speed of contemporary high performance liquid chromatography mass spectrometry (HPLC/MS) analysis. The quantitative analysis of a test analyte (Ziprasidone) in plasma demonstrates the utility and throughput of membrane SPE in combination with HPLC/MS. The results obtained with the current automated procedure compare favorably with those obtained using solvent and traditional solid phase extraction methods. The method has been used for the analysis of numerous drug prototypes in biological fluids to support drug discovery efforts. PMID- 9204579 TI - Kinetic analysis and multiple component monitoring of effectors of adenylyl cyclase activity by quantitative fast-atom bombardment mass spectrometry. AB - The enzyme adenylyl cyclase catalyses the conversion of adenosine 5'-triphosphate (ATP) to adenosine-3',5'-cyclic monophosphate (cyclic AMP), and is an important pharmaceutical target. Quantitation of this enzyme's activity has been carried out by positive-ion fast-atom bombardment mass spectrometric analysis of the enzyme incubation mixture after the reaction has been terminated. The kinetic data obtained are in good agreement with those obtained by the conventional radiometric assay, and this mass spectrometry-based assay offers the facility to monitor the turnover of several components of the incubation simultaneously. This is utilized to study the relative efficiencies of two ATP-regenerating systems, three phosphodiesterase inhibitors and two modified substrates, and to monitor the uptake and conversion of two competing substrates, adenosine 5' triphosphate and 2'-deoxyadenosine-5-triphosphate, to cyclic AMP and to cyclic deoxyAMP, respectively. PMID- 9204580 TI - Sequence database searches via de novo peptide sequencing by tandem mass spectrometry. AB - A method is described for searching protein sequence databases using tandem mass spectra of tryptic peptides. The approach uses a de novo sequencing algorithm to derive a short list of possible sequence candidates which serve as query sequences in a subsequent homology-based database search routine. The sequencing algorithm employs a graph theory approach similar to previously described sequencing programs. In addition, amino acid composition, peptide sequence tags and incomplete or ambiguous Edman sequence data can be used to aid in the sequence determinations. Although sequencing of peptides from tandem mass spectra is possible, one of the frequently encountered difficulties is that several alternative sequences can be deduced from one spectrum. Most of the alternative sequences, however, are sufficiently similar for a homology-based sequence database search to be possible. Unfortunately, the available protein sequence database search algorithms (e.g. Blast or FASTA) require a single unambiguous sequence as input. Here we describe how the publicly available FASTA computer program was modified in order to search protein databases more effectively in spite of the ambiguities intrinsic in de novo peptide sequencing algorithms. PMID- 9204581 TI - Early defibrillation by first responding ambulance personnel. PMID- 9204582 TI - The effect of semi-automatic external defibrillation by emergency medical technicians on survival after out-of-hospital cardiac arrest: an observational study in urban and rural areas in Belgium. AB - The introduction of semi-automatic external defibrillators (SAEDs) allowed emergency medical technicians (EMTs) to deliver electroshocks in cases of out-of hospital ventricular fibrillation (VF) or ventricular tachycardia (VT), often many minutes before the arrival of the mobile intensive care unit (MICU) team. In this observational study we report on the results obtained by the EMTs from the fire departments of Gent, Aalter and Brugge. In Gent, an SAED project started in May 1991. By December 1995, the SAED's electrodes had been attached in 367 cardiac arrest patients. The first rhythm detected by the device was asystole or electromechanical dissociation (EMD) in 241 patients (66%): only 5 of these patients survived to hospital discharge (2%). In the remaining 126 VF/VT cases (34%) the survival rate was 21% (26/126). In 14 of these 26 patients the shock(s) delivered by the EMTs restored spontaneous circulation before the arrival of the MICU team, with only venous cannulation and/or intubation being performed by the MICU team. In 4 other VF patients, the shock(s) delivered by EMTs converted the VF, with the MICU team successfully taking care of VF/VT relapses or postcountershock EMD. In the remaining 8 VF/VT cases, only the MICU attempts could resuscitate the patient. The SAED project in Aalter was set up in April 1993. By December 1995, care was taken for only 21 patients. None of the 4 VF/VT patients and the 17 asystole/EMD patients survived. In Brugge, there were 240 cardiac arrest cases treated with SAED between January 1991 and December 1995. Among the 89 VF/VT cases, there were 20 survivors (22%): 8 cases survived thanks to SAED shock(s) delivered by EMTs, in 3 cases survival was due to the combination of SAED shock(s) by EMTs and extensive ALS treatment by the MICU team, and in 9 cases restoration of spontaneous circulation was only obtained after application of ALS techniques by the MICU team. This observational study seems to show a beneficial effect of the introduction of SAED in Gent and Brugge. In Aalter the number of treated cases is tool low to draw conclusions. Anyhow, the global survival rate in the three areas remains low. Therefore, more efforts are needed to strengthen the other links of the chain of survival (early access to the emergency medical services-system, early basic cardiopulmonary resuscitation and early advanced life support. PMID- 9204583 TI - Comparative study of antiphospholipid antibody detection in eleven Belgian laboratories. AB - Twenty-six plasma samples have been sent to 11 different Belgian laboratories in order to detect the presence of antiphospholipid antibodies, either by immunological methods and/or by coagulation tests. A good concordance between laboratories was observed for coagulation tests. Laboratories using detection tests and performing mixing procedures and neutralisation procedures displayed the highest sensitivity as compared with laboratories which did not perform one of these two latter procedures. The concordance between laboratories for the immunological methods was much worse as compared with coagulation tests. This may be attributable either to an intrinsic problem of the immunological tests or to a selection bias due the fact that the plasmas used in this study were selected in coagulation laboratories only where the chance to find a lupus anticoagulant positive/ELISA antiphospholipid negative sample is high. PMID- 9204584 TI - Current approaches to histocompatibility testing. A short overview. PMID- 9204585 TI - Allergenic proteins in different brands of latex and synthetic medical examination gloves. AB - The steady increase in anaphylactic reactions to latex medical gloves has raised increasing awareness in the medical community. Even gloves claimed to be hypo allergenic still may contain substantial amounts of IgE-binding proteins. We have studied non-powdered latex and synthetic examination glove extracts for their allergenicity using the immunoblot technique. Protein levels varied considerably among glove extracts and the amount did not always correlate with the presence of allergenic proteins. IgE binding proteins were found in 2 of the 7 powder-free latex glove brands. Synthetic glove extracts did not contain allergens. The study demonstrates that immunoblot analysis is a useful technique in order to select gloves with a minimal risk to raise IgE antibodies. PMID- 9204586 TI - Usefulness of induced sputum analysis in pulmonary diseases. AB - In this study, we analysed the cellular component of induced sputum in healthy control subjects (n = 30), asthmatics (n = 44), patients suffering from COPD (n = 15), pulmonary tuberculosis (PTB) (n = 14) and healthy steel workers (HSW) (n = 14). Sputum was induced by inhalation of hypertonic saline (NaCl 5%) for 20 min. When compared to the healthy control group, all the disease groups as well as the one of healthy steelworkers exhibited significantly higher total sputum cell counts. Analysis of the differential cell counts showed that there was a significant increase in % eosinophils in asthmatics, in % neutrophils in COPD, asthmatics, tuberculosis and healthy steelworkers and in % lymphocytes in pulmonary tuberculosis. Our study illustrates the feasibility and the possible clinical application of induced sputum analysis in several pulmonary diseases and shows how this technique could be useful in assessing airway inflammatory processes in subjects exposed to industrial pollutants. PMID- 9204588 TI - Impredictable susceptibility for cephalosporins in penicillin resistant Streptococcus pneumoniae. PMID- 9204587 TI - Severe community-acquired pneumonia caused by atypical organisms. AB - Three cases of community-acquired pneumonia (CAP), requiring intensive care admission, are presented. The clinical picture of a "typical" bacterial pneumonia in the three patients led to an initial empirical treatment with amoxicillin clavulanic acid or 2(nd) generation cephalosporins. The treatment had to be changed in all three because of clinical failure. Erythromycin was added to the therapy with good clinical evolution. Serology confirmed atypical organisms to be responsible. Only the chest X-ray might have suggested an "atypical" or a "viral like" agent. A proposition is made for an empirical combination of antibiotics in severely ill patients with CAP with more than unilobar consolidation. PMID- 9204589 TI - Historical development of Croatian radiology. AB - The history of the development of Croatian radiology is described, from the first beginnings to the present. Very soon after Roentgen's discovery in 1895, not only communications and informative lectures, but also the first x-ray device appeared in Zagreb and then in many other towns in Croatia. The first specialists in roentgenology and x-ray technician were educated at Professor Holzknecht's Roentgenology Institute in Vienna. After the opening of the University School of Medicine in Zagreb in 1917/1918, the Chair of Roentgenology, headed by Professor L. Popovic, was founded as early as 1922. Here the first specialization for physicians was organized. After the second world war, with the foundation of Medical Schools in Rijeka, Osijek and Split, Croatian radiology steadily progressed. Autonomous departments and institutes for diagnostic and therapeutic radiology were organized at all large medical centers. Thanks to gynecologists, ultrasound has not only developed at radiologic units, but also at departments of gastroenterology, orthopedics, pediatrics and others. Today, there are many units for interventional ultrasonography throughout Croatia. Radioisotope units have also been organized, but only in the main medical centers, rarely in small ones. At present, there are five radiotherapy and oncology institutions in Zagreb, and one in Rijeka, Split and Osijek. Interventional radiology has been well organized since the 80s, and the First Congress of Diagnostic and interventional Radiology was held in Rijeka in 1994. In conclusion, there is no doubt that the Croatian radiology keeps pace with the international advances in the field. PMID- 9204591 TI - Prevalence and significance of hepatitis C virus (HCV) genotypes in anti-HCV positive patients in northwest Croatia. AB - Hepatitis C virus (HCV) transmission is related to blood transfusion and transmission in hemodialysis (HD) units. The aim of the study was to investigate the distribution of HCV genotypes and routes of HCV transmission in three groups of anti-HCV positive patients in north-west Croatia. A total of 111 patients were studied. Patients were classified into three groups: 45 HD patients with a low percentage of anti-HCV positivity (group 1), 60 HD patients treated at another HD unit and with high percentage of anti-HCV positivity (group 2), and six anti-HCV positive patients with chronic hepatitis (group 3). Most of the HD patients were treated during the sama shift, but with separate equipment. Serum HCV RNA was detected by reverse transcription-polymerase chain reaction (RT-PCR). Genotyping of HCV isolates was performed with a line-probe assay. Patients of groups 1 and 2 did not show significant differences with regard to the clinical profile. In group 1, all anti-HCV positive patients were RT-PCR positive and all of them were infected with the same genotype (genotype 3). In group 2, 89% of anti-HCV positive patients were RT-PCR positive and all infected with the same genotype (genotype 1 b). In group 3, 50% anti-HCV positive chronic hepatitis patients were RT-PCR positive. Two of them were infected with genotype 1 b and one with genotype 4. The homogeneity of HCV genotypes in the patients from both HD units (groups 1 and 2), seemed to indicate nosocomial transmission of HCV, whereas viremia was found to be related to blood transfusion in all group 3 patients. The exact mechanism involved in the transmission of HCV in HD units remains to be discovered. PMID- 9204590 TI - Follow-up of patients with gastrointestinal cancer by tumor marker determination. AB - In addition to clinical examination and various diagnostic procedures, patients with gastrointestinal cancer (GIC) were also monitored by tumor marker (TM) determination. In total, 202 patients with GIC were postoperatively followed-up. According to in vivo diagnostic procedures, there were 133 (66%) patients without metastases, 63 (31%) patients with distant metastases, 48 (76%) of them with liver metastases, and six (3%) patients with local recurrences. During the 1990 1995 period, they were followed-up by serum TM concentration measurements on 1-8 occasions. At the time of initial diagnosis, TM were determined in a varying percent of the patients: CA 19-9 (100%), TPA (54%), CA 72-4 (49%), IAP (41%), CEA (41%) and AFP (25%). In the group of patients without metastases, the percentage of normal TM values ranged between 73%-100%, and TM sensitivity between 0%-86%. No ideal TM has as yet been discovered either for any malignant disease or for patients with GIC. Therefore, in the follow-up of patients with GIC we suggest the concentrations of at least two instead of only one TM (CA 19-9 and TPA or CEA) to be determined. It is also difficult to choose the best TM among numerous TM kits available on the market and to keep using it during a longitudinal follow up study. PMID- 9204592 TI - Nucleolar organizer region in transitional cell carcinoma of the bladder. AB - The silver staining technique that identifies NORs (nucleolar organizer regions) associated proteins was used for examining changes in nucleolar organizer region numbers in transitional cell carcinoma of the bladder. This method reveals NORs as black dots in the cell nuclei, and the number of NORs per cell (NORs/cell) has been taught to be related to cellular activation and to be a possible predictor of clinical outcome. A hundred specimens of transitional cell carcinoma of the bladder divided into four histologic grades (Ash classification) with 25 specimens each, and 25 specimens of normal bladder mucosa were analyzed. It has been demonstrated that the amount of protein synthesized by carcinoma varies according to its histologic grade, i.e. the higher histologic grade, the greater the NORs/cell number for each histologic grade. The characteristic mean NORs/cell SD was determined (GI 4.85 +/- 0.82, GII 5.94 +/- 1.42, GIII 8.54 +/- 1.01 and GIV 8.72 +/- 1.42), among which statistically significant differences were found. For similar histologic grades the characteristic mean NORs/cell +/- SD showed no statistically sex and age related differences. In some cases the mean NORs/cell for the examined carcinoma did not match the characteristic mean NPRs/cell +/- SD for the respective histologic grade. PMID- 9204593 TI - Dialyzer reprocessing with peroxyacetic acid as sole cleansing and sterilizing agent. AB - In a prospective study, the effectiveness of a newly available peroxyacetic acid solution (Dialox) as a cleansing and sterilizing agent for the reuse of conventional hollow-fiber hemophan dialyzers was evaluated. The effects of reprocessing dialyzers with peroxyacetic acid on leukocyte and platelet counts, dialyzer performance, acute intradialytic symptoms and cost effectiveness were studied. A total of 250 sessions using new dialyzers and 3,227 sessions employing reused dialyzers were monitored. Dialyzers were withdrawn after a maximum of 21 sessions, and the mean number of uses was 13.9 +/- 5.1. It was found that peroxyacetic acid reprocessing of hemophan dialyzer membranes improved hemodialysis leucopenia and thrombocytopenia. It was also proved that, compared to new ones, the reused dialyzers were associated with significantly fewer intradialytic symptoms. Clearances of small molecules and ultrafiltration rate after multiple dialyzer reprocessing remained within the acceptable limits. The duration of hemodialysis has been prolonged 30 min after the 10th dialyzer reuse and dialysis adequacy remained unaffected. About 100,000 DEM were saved during the one year due to the reuse procedure. The authors conclude that automated dialyzer reprocessing with peroxyacetic acid as a sole cleansing and sterilizing agent is a safe, efficacious and money-saving method. PMID- 9204594 TI - Another therapeutic schedule in eradication of Helicobacter pylori. AB - In this study, the efficacy and tolerability of two different therapeutic schedules in eradicating Helicobacter pylori and healing duodenal ulcer were evaluated. The study included 60 patients with duodenal ulcer and Helicobacter pylori infection. They were randomly allocated to either of two groups: group 1 (N = 30) received omeprazole 20 mg for 28 days, amoxicillin 3 x 500 mg for 7 days and metronidazole 3 x 500 mg for 5 days, and group 2 (N = 30) received omeprazole 20 mg for 28 days, ACA (amoxicillin 500 mg plus clavulanic acid 125 mg) 3 x 625 mg for 7 days and metronidazole 3 x 500 mg for 5 days. Endoscopic examination, bioptic urease test and histologic examination were performed before, and 30 and 90 days after the treatment. Endoscopic examination was also performed one month after the beginning of the treatment, when healing of duodenal ulcer was observed in 90% (27/30) of the group 1 patients and in 93.3% (28/30) of the group 2 patients. The Helicobacter pylori eradication achieved in group 1 and 2 was 76.7% (23/30) and 83.3% (25/30), respectively. Side effects were present in 20% (6/30) of the group 1 patients and in 23.3% (7/30) of the group 2 patients. Side effects were mild and did not require interruption of the treatment. A higher rate of eradication was achieved in group 2 than in group 1, but the difference was not statistically significant. PMID- 9204595 TI - Sensorimotor stimulation of comatose patients. AB - Coma is a state of unconsciousness from which the patient cannot be aroused. The causes of coma are classified into structural and nonstructural disorders. The depth and duration of post-traumatic consciousness disorder are a sensitive and reliable indicator of the severity of head trauma. The social prognosis aggravates with age and duration of unconsciousness. Early sensorimotor stimulation by adequate series of stimuli paves the road for new collaterals of the weakened and lost neurons, and for their connection. In such a way. the entire system of structures required for successful regeneration and reorganization of functions in the damaged parts of the brain is maintained in a relatively functional tone. This is substantiated by two examples taken from practice, presented as brief case reports. PMID- 9204596 TI - The effect of hollow-fiber dialyzer Plivadial Altra-Flux 140 on beta-2 microglobulin removal. AB - A thirty-fold or even greater increase in plasma beta-2-microglobulin (beta 2M), which is commonly found in end-stage renal disease (ESRD) patients on long-term hemodialysis (HD), is most likely a consequence of the inability of the dialysis procedure to remove the dally production of beta 2M. In the present study, a newly developed high-flux membrane composed of cellulose diacetate (CDA) (dialyzer Plivadial Altra-Flux 140, Pliva, Zagreb, Croatia) was evaluated with regard to beta 2M removal capacity during HD in 8 stable ESRD patients. Thera was a drop in the plasma beta 2M concentration (-19.8 +/- 8.4) with a clearance of 22.7 +/- 9.2 ml/min (QB = 250 ml/min, QD = 600 ml/min). Accordingly, the sieving coefficient (SC) was found to be 0.37 +/- 0.1 at 60 min after the start of HD. The CDA membrane was able to remove 100.5 +/- 30 mg of beta 2M during a 4-hour HD session. This data demonstrate an increased percentage removal of beta 2M and significantly decreased postdialysis plasma concentrations of beta 2M which is a potential factor in the development of dialysis-related amyloidosis (DRA). PMID- 9204598 TI - A hundred and fifty years of the general hospital in Nova Gradiska (1846-1996). PMID- 9204597 TI - Prostatic polyp in the prostatic urethra. AB - The authors report on a case of prostatic polyp in the urethral bulb of a 35-year old man. The presence of this lesion in the urethral bulb is quite unusual and may be overlooked on cystocopic evaluation. PMID- 9204599 TI - On the 50th anniversary of Acta Medica Croatica. PMID- 9204602 TI - Infectious diseases in the 21st century. AB - Infecto-contagious diseases in the twenty-first century with respect to precedent will see themselves deprived of smallpox, dracunculiasis and very probably of paralyzing poliomyelitis. Vaccination-preventable diseases, such as measles, whooping cough, diphtheria, tetanus, rabies, some forms of meningitis, yellow fever and episodes of disseminated tuberculosis will greatly diminish in their rates of morbi-lethality; the elimination of some, and the eradication of measles, are expected. Other diseases such as diarrhea (including cholera), geo helminthiasis, some severe respiratory tract infections and the majority of vector-transmitted infectious diseases will decrease due to improvements in potable water services, drainage, sanitary food control, living quarters, and individual and community anti-vector action. Leprosy, onchocerciasis and several parasitoses will be controlled by the available antimicrobial drugs. Infectious diseases will continue to be an important health problem due to: Reduction in the immunocompetence resulting from the aging of the population, chemotherapies necessary for neoplasms, and autoimmune pathology and the survival of persons with primary immunodeficiencies; lifestyles prone to infectious pathology, such as mega-city urbanization, children in day care centers, industrialized foods, intravenous drug addiction, sexual liberation, global commerce, and tourism; antibiotic-multiresistant microbial flora; environmental disturbances as a result of global warming, deforestation, the settling of virgin areas, dams, the large scale use of pesticides, fertilizers and antimicrobials, and natural/social disasters generators of poverty, violence and deprivation will result in emergence or re-emergence of infectious diseases already controlled in the past. PMID- 9204603 TI - Some consequences of ozone exposure on health. AB - The reaction of ozone with polyunsaturated fatty acids from the surfactant factor and pulmonary epithelial cells produces different reaction products which can cross the alveolar-capillary barrier and reach distant structures. Although only a few papers claim extrapulmonary changes in animals exposed to this gas, some neurological deficits, such as complaints of fatigue, lethargy, headache in humans, as well as significant disarrangements in the sleep pattern related to biochemical changes in the brain have been referred to in animals exposed to ozone. In the present review, the molecular configuration and the reaction of ozone at different lung levels are related to impairment at the respiratory and blood systems, in order to elucidate the mechanisms by which this gas or its reaction products, such as free radicals, prostaglandins and others can cross the alveolar-capillary and hemato-encephalic barriers, and to explain those neurological effects. PMID- 9204604 TI - Giardia duodenalis: analysis of humoral immune response in experimentally infected gerbils (Meriones unguiculatus). AB - In this work, we have analyzed the humoral immune response in Mongolian gerbils infected with Giardia duodenalis trophozoites of strains P-1 and WB. The course of infection in the animals was assessed by monitoring cyst shedding in feces, and serum samples were collected at weekly intervals to measure antibody levels by ELISA. Parallel studies were carried out to determine the patterns of total and surface antigens of the parasite recognized by antibodies using Western blot and radioimmunoprecipitation (RIP) assays with the use of homospecific enzyme conjugates. Typical patterns of cyst shedding were observed in the infected animals and cyst numbers per gram of feces were consistently higher in gerbils infected with WB strain. Antibody levels to G. duodenalis antigens were observed by week 2 post-infection and were still detectable 4 months after infection. G. duodenalis antigens showed a complex but quantitatively and qualitatively different recognition pattern by infection-induced antibodies in Western blot assays which related to infecting strain. However, RIP assays showed a more restricted and common pattern of recognition of surface antigens from either strain. Taken together, the data obtained in this study provides further information regarding direct comparisons among infecting strain, patterns of infectivity, and host immune response toward G. duodenalis antigens in the gerbil model. PMID- 9204605 TI - Increased yield of F VIII from pooled plasma vs. single-donor plasma in the production of cryoprecipitates. AB - Coagulation factor VIII:C yield was studied in two types of cryoprecipitates. The first group contained products from single-donor plasma units. The other group contained cryoprecipitates which were produced from pooled plasma. The volume of plasma/bag was not different between the two groups, but both the yield and the total content of F VIII:C in cryoprecipitates were significantly different. The yield of F VIII:C was higher (+20% in relative terms) in cryoprecipitates produced from pooled plasma, resulting in higher potency of such products. The positive effect of plasma pooling on the recovery of F VIII:C might be a result of reassembly of factor VIII subunits of different individuals in the plasma pools. The findings may have a role also in large-scale production of F VIII concentrates. PMID- 9204606 TI - Limits of variation of fiber distribution in the sural nerve of man. AB - Although the sural nerve is the most extensively studied nerve in man, there is a dearth of data regarding the normal variations in the size-frequency distribution of axons in normal subjects; criteria for assessing the normality of a given individual are not available. Therefore, in everyday practice, the surgical pathologist may meet with difficulty in interpreting the biopsy of one particular individual, in whom the distribution is slightly different from the curves published. The object of this work is to detect the normal limits of variation in the distribution of diameters of myelinated and unmyelinated fibers in normal subjects and to establish the criteria that permit the calculated curves to be used in everyday clinical practice. Normal sural nerves of 19 patients were analyzed. Ages ranged between 18 months and 55 years. Morphometric analysis was performed with the Histoscan X automatic image processing analyzer, and, for statistical analysis, mixtures of lognormal distributions were fitted and tested with Pearson's statistics. Nerves of three diabetic patients were used for testing the method. They were clearly classified as abnormal. The curves, therefore, have been proven useful for everyday surgical pathology practice. PMID- 9204607 TI - Tachyphylaxis and sensitization to nicotine-induced tachycardiac and pressor effects after nicotine infusions. AB - This work examined the effects of nicotine on mean arterial pressure and heart rate in non-anesthetized spinal rats. Nicotine (200 mg/kg) was administered as a single bolus, as infusions lasting 7.5, 15 or 30 min, and as a post-infusion bolus. A nicotine bolus increased pressure and rate. These effects were less marked as the rate of infusion decreased. The infusions affected differentially the effects of a subsequent bolus. Thus, while tachycardia was decreased, the blood pressure rise was increased. An initial transient bradycardia was observed after bolus administration, but not during infusions; this effect was unchanged after post-infusion boluses. Pharmacological analysis indicated that tachycardia and bradycardia were predominantly due to ganglionic stimulation, while adrenal and sympathetic nerve catecholamine release played a major role in the pressor response. These results indicate that slow nicotine infusions do not induce tachyphylaxis for all of the cardiovascular effects of a subsequent bolus, and that development of acute tolerance appears to depend on the mechanism of action of the response. PMID- 9204608 TI - Bacterial resistance to antibiotics in acute respiratory infections (ARIs). AB - In this review article, we make suggestions on how to approach the increasing problem worldwide of bacterial acute respiratory infections resistant to antibiotics. After a brief description of the main mechanisms of bacterial resistance, i.e., enzymatic inactivation by beta-lactamases, reduction in the permeability of the outer membrane and the development of PBPs that have decreased affinity for the antibiotic, we analyze documented experiences on the response to different groups of antibiotics (beta- lactam antibiotics, cephalosporins, carbapenems and quinolones), of the most commonly isolated bacteria from invasive respiratory infections (Haemophilus influenzae, Streptococcus pneumoniae and Moraxela (Branhamella) catarrhalis. Antimicrobial agent susceptibility in vivo and in vitro testing and the correlation of their results provide the basic information for the adoption of adequate policies and strategies for better use of antibiotics in bacterial respiratory infections; proper surveillance would allow to make intelligent changes in such a policy. Standardized recommendations for clinical practice on the use of antibiotics could be misleading, iatrogenic, and could complicate the resistance problem. To prevent and control the rise and spread of bacterial resistance, an interdisciplinary approach is needed. PMID- 9204609 TI - Changes in human serum antioxidant capacity and peroxidation after four months of exposure to air pollutants. AB - Twenty-one adult volunteers (aged 27-32 years), who had been living in Mexico City for four continuous months (physicians working as fellows) were studied the first and sixteenth week of their stay in order to learn the effects of the pollutants contained in Mexico City's atmosphere on some serum biochemical parameters. The activity of serum superoxide dismutase (SOD) decreased after 16 weeks in comparison with the values obtained the first week (109.6 to 56.9 mU/mg protein; 50% less). In contrast, the inhibitory capacity of serum vs. induced in vitro lipoperoxidation increased in relation to the length of stay (22%). The serum levels of thiobarbituric-reactive material also decreased in almost 30% (from 6.10 to 4.12 nmol). The other lipoperoxides measured were unchanged (chromolipids and diene conjugation). We propose that this may be as a result of the adaptative capacity of the human organism, within a pollutant atmosphere in which the ozone levels might participate in a decrease of SOD activity during chronic exposure, to air pollution. PMID- 9204611 TI - Effect of chemotherapy on thyroid hormone concentration in patients with malignant hematologic diseases. AB - Malignant hematologic diseases are severe illnesses for which complex forms of treatment are used and to which a great variety of metabolic changes are anticipated. Both lymphomas and leukemias, as well as chemotherapy, can induce abnormalities in thyroid hormone metabolism without overt disease, thus leading to what is known as euthyroid sick syndrome (ESS). In the present report, 25 patients with lymphomas and leukemias were studied to evaluate the effect of chemotherapy on thyroid hormone concentration. After chemotherapy, the most frequent and significant alteration was a decrease in serum triiodothyronine concentration. PMID- 9204610 TI - Cytogenetic findings in 303 Mexican patients with de novo acute myeloblastic leukemia. AB - In this report we show the chromosomal changes seen in a group of 303 Mexican patients with de novo Acute Myeloblastic Leukemia (AML). Two hundred forty-two patients were diagnosed and treated at two hospitals affiliated with the Instituto Mexicano del Seguro Social (IMSS). These are the Centro Medico Nacional Siglo XXI and Centro Medico La Raza Hospitals; the remaining 61 patients were diagnosed and treated at the Hospital General de Mexico (HGM). Clonal abnormalities were detected in 75.6% of the patients; this result agrees with what has been reported in other large series of AML studies. The incidence of changes per hospital was similar in patients from the IMSS hospitals (72-75%), while an increase was seen in patients from the HGM (85.2%). The chromosomal changes seen in this study in order of frequency were: t(15;17)[18.8%], t(9;22)[9.2%], miscellaneous chromosomal changes (mainly rearrangements of chromosomes 1,2,3,12y17)[8.2%], abnormalities of 16q22 [7.3%], t(8;21)[6.3%], 7/del(7q)[5.6%], t(6;9)[5.3%], and abnormalities of 11q23 [4.6%]. We reported an increase in the incidence of certain types of chromosomal changes seen in cases of AML, in comparison with reports from other countries. These differences could be due to methodological variations, although ethnic, socioeconomic and nutritional differences must not be disregarded. We support this finding when comparing distribution of changes in the population of patients seen in the IMSS hospitals with those from the HGM; the main difference lies in the socioeconomic level. PMID- 9204612 TI - Frequency of intestinal parasites in adult cancer patients in Mexico. AB - Approximately 28% of the Mexican population has intestinal parasites. Oncologic patients receiving chemotherapy should have a coproparasitoscopic study to avoid disseminated parasitic infections. The frequency of intestinal parasites, including Cryptosporidium and Isospora, was evaluated in 100 diarrheic (DS) and 100 formed stools (FS) from adult patients recently diagnosed with cancer, using wet mounts stained with Kinyoun, saccharose and ZnSO4 procedures stained with Lugol's iodine. Seven patients with DS and three with FS had more than one parasite. Pathogenic intestinal parasites were seen in 26% of DS and 15% of FS. Of the frequent parasites, Entamoeba histolytica was found in 12 DS and in 2 FS (p = 0.01), Giardia lamblia in three DS and six FS and Hymenolepis nana in eight DS and 10 FS. Other pathogenic parasites were found only in DS: Cryptosporidium sp. in five patients, Ascaris lumbricoides in two, Strongyloides stercoralis in two and Isospora sp. in one. Cryptosporidium and Isospora were only identified by wet mounts stained with Kinyoun while other parasites were identified by flotation procedures. Since six (3%) of our patients had coccidia, the laboratory must perform special techniques for their detection. In epidemiologic settings where there is a high prevalence of intestinal parasitic infections the coproparasitoscopic studies should be performed and antiparasitic treatment provided before starting chemotherapy. PMID- 9204613 TI - Hypertonic-hyperosmotic solution modifies myoglobin levels in early reperfusion after ischemic cardiac arrest. Experimental model. AB - To evaluate the effect of reperfusion with hypertonic-hyperosmotic solution, cardiectomy was performed in 25 New Zealand white rabbits. Seven isolated hearts were submitted to 30 min of global ischemia and reperfused with oxygenated buffer for 60 min. Myoglobin and isoenzyme MB of creatine kinase concentrations were each measured in the effluent 15 min, and values were correlated (r = 0.5011, p = 0.015). After this procedure, 18 isolated hearts were randomized in two groups. Hearts of group I were reperfused with hypertonic-hyperosmotic solution (NaCl 7.5% dextran 60,000 MW) diluted in oxygenated buffer, and group II with oxygenated buffer. Myoglobin and coronary flow were measured in both groups, group I showed lower levels of myoglobin (p = 0.0069) and higher coronary flow (p = 0.028) than group II. In conclusion, changes in myoglobin concentration in the heart effluent are more sensitive than changes in isoenzyme MB of creatine kinase; thus, evaluation of this parameter may be useful in the detection of ischemia reperfusion injury. Additionally, hypertonic-hyperosmotic solution improves the coronary flow and has a protective effect against ischemia reperfusion injury. PMID- 9204614 TI - Mortality associated with systemic candidiasis in children. AB - The purpose of this study was to determine factors associated with an increased risk of mortality due to systemic Candida infections in children hospitalized at our tertiary care facility. A total of 71 cases of Candida bloodstream infections were identified over a 2-year period. The attack rate was 47 cases of candidemia per 10,000 discharges and the case fatality rate was 46.5%. Sixty-one cases occurred in infants under 2 years; 27 were newborns (38%). Using logistic regression analysis, we evaluated the independent effects of potential risk factors for death due to candidemia. Three factors were associated with the subsequent risk for death due to systemic candida infection: malnutrition (OR = 4.3; 95% CI 1.2-14.8), prior surgery (OR = 3.8; 95% CI 1.2-13.2), and the number of days between the first positive candida blood culture and the onset of antifungal treatment (OR = 1.12; 95% CI 1.06-1.25). Newborns showed an almost three times greater risk of death due to candidemia as compared to other age groups, but this association was only marginally significant (OR = 2.8; 95% CI 0.9-9.3). There was no difference in the rate of candidemia between the 2 years of the study; however, the observed mortality declined significantly from 65% in year one to 20% in year two (p = 0.02). The major finding of this study was to observe that for every day treatment was delayed the risk of death increased significantly. Thus, this study provides support for empirical antifungal therapy early in the course of suspected systemic candidiasis in order to improve survival among children. PMID- 9204615 TI - B-lineage acute lymphoblastic leukemia of childhood. An institutional experience. AB - A total of 119 children (1990-95) with acute lymphoblastic leukemia (ALL) B lineage either CD10+ or CD10- were registered into a single non-randomized chemotherapy protocol. Only untreated patients with standard risk were included in the study. Their ages ranged from 1.8-10 years with a mean of 5.1 years. There were 82 (68%) children with early pre B-All, 35 (29%) with pre B-ALL and 2(1.6%) with transitional pre B-ALL (p < 0.00001). The patients were divided according to CD10 reactivity, either + (94 children) or -(25 patients). The event-free survival (EFS) at 60 months for the CD10+ children was of 78% (alive 73/94), while for the CD10- was 71% (alive 18/25) (p = 0.6) and 74% for both groups. The factors that influenced favorably the survival in the CD10+ group were the age between 3 to 5.99 years (p < 0.00001), sex (either male or female), leukocyte count between 10-24.9 x 10(9)/l (p < 0.00001), LDH under 300 U/I (p < 0.00001) and L1 bone marrow cytomorphology (p < 0.00001). In the CD10- patients, the EFS was favorably influenced by the female sex (p = 0.04), leukocyte count under 10 x 10(9)/l (p = 0.05) and LDH < 300 U/l (p = 0.02). CNS infiltration was documented in 4.2% (5/119). Mortality secondary to chemotherapy was seen in 7%. In conclusion, this is the first large series in Mexican children with B-lineage ALL published. Because of the relatively small number of patients in each group (pre B and transitional pre B), all the patients in the current series were treated alike. When the 119 patients were divided only on the basis of CD10 reactivity, the EFS for both groups (CD10+ and-) was similar; therefore, the reactivity to CD10 has no prognostic value in this type of ALL. PMID- 9204616 TI - Treatment of active gastroesophageal variceal bleeding with terlipressin or hemostatic balloon in patients with cirrhosis. A randomized controlled trial. AB - Gastroesophageal variceal bleeding due to portal hypertension should be treated by endoscopic sclerotherapy. This procedure, however, has some limitations. It has been established that vasoactive drugs are effective for controlling active variceal bleeding. We report the results of a randomized controlled trial comparing terlipressin to hemostatic tube (Linton-Michel tube) for the treatment of bleeding gastroesophageal varices in cirrhotic patients. Thirty-seven cirrhotic patients with a total of 40 episodes of gastroesophageal variceal bleeding were included in this trial. Patients were randomly assigned to intravenous terlipressin or Linton-Michel tube (LM tube), for 24 h. During this period, hemostasis was defined as obtaining of hemodynamic and hematocrit stabilization and/or absence of hematemesis or melena. Bleeding recurrence was assessed during a 1-month period after treatment. Twenty bleeding episodes were treated with terlipressin (Group I) and 20 with LM tube (Group II). Both groups of patients were similar in age, sex distribution, etiology of cirrhosis and degree of hepatic insufficiency. Bleeding was controlled in 70% of patients from Group I and in 95% from Group II (p < 0.05) during treatment. Bleeding recurred in 14% of patients in Group I vs. 36% in Group II 1 week following the treatment (p > 0.05) and in 16.6% in Group I vs. 83.3% in Group II 1 month after treatment (p < 0.05). Complications were more frequent in Group II than in Group I (65 vs. 15%, p < 0.05). Mortality rate was similar in both groups 1 month after treatment. In conclusion, hemostatic tubes were superior to terlipressin for the control of active gastroesophageal variceal bleeding within the first 24 h. Complications and bleeding recurrence were more frequent in patients treated by hemostatic tube within a period of 1 month after treatment. Mortality rate was similar in both groups of patients. PMID- 9204617 TI - Usefulness of cryohemolysis test in the diagnosis of hereditary spherocytosis. AB - The clinical suspicion of hereditary spherocytosis (HS) must be confirmed at the clinical laboratory. The osmotic fragility test (OFT) and the autohemolysis test (AHT) are the worldwide accepted assays to establish a definite diagnosis of HS; however, they have some disadvantages. We describe herein our experience with the cryohemolysis test (CHT) as a tool to confirm the HS diagnosis. We included four groups of subjects, namely, patients with clinical HS, patients with mechanical heart valve prosthesis, malignant hematological diseases and healthy blood donors. CHT was carried out in all the groups, while OFT and AHT only in the HS patients and healthy individuals. OFT and AHT were performed according to previously described techniques. CHT was performed using red blood cells incubated in a hypertonic solution, preheated for 10 min and then transferred to an ice bath for an additional 10 min. The resulting cryohemolysis was determined measuring the free hemoglobin in the sample. There were no differences among the groups in terms of general characteristics. All HS suspicious patients had a positive OFT and AHT. CHT was positive in all patients from the HS group but in none of the subjects from the control groups (p < 0.001). We found that CHT is a faster and easier-to-perform assay compared with OFT and AHT. Moreover, using CHT, the zone between normal and abnormal results is wider than OFT or AHT. We propose 0.7 to 11% hemolysis as reference values for CHT. PMID- 9204618 TI - Usefulness of fractional excretion of sodium in critically ill pre-term newborns. AB - The purpose of this prospective study was to measure the fractional excretion of sodium (FENa) in critically ill pre-term newborns (PTNB) in order to determine its cut point in the diagnosis of acute renal failure (ARF). This study included 52 newborns and was conducted from May, 1994 to May, 1995. Patients were divided in two groups: patients without ARF in group A (n = 47) and patients with ARF in group B (n = 5). No statistically significant differences were found in birth weight, extrauterine life span, serum sodium levels, urine creatinine and urinary volumes between the two groups, but there was a difference in gestational age, urinary sodium concentration and serum creatinine levels. Sensitivity and specificity were determined, and the critical level of FENa was 4% or greater for ARF diagnosis. The average FENa value for Group A was 1.4 +/- 1.4% with a median of 0.92%. In Group B, average FENa was 6.9 +/- 2.9% with a median of 8.5% (P < 0.001). We conclude that FENa is a valuable tool for the assessment of renal function in critically-ill PTNB, in spite of all other factors present in this population that could modify its values. PMID- 9204619 TI - Diagnostic value of the copper/zinc ratio in digestive cancer: a case control study. AB - The aim of this study was to assess the accuracy of the copper/zinc ratio (Cu/Zn ratio) in the evaluation of a large group of patients with digestive cancer compared to gender and age-matched control subjects. A total of 282 patients was studied and separated into three groups: group I (n = 75), patients with digestive cancer, group II (n = 112), patients with benign digestive disease, and group III (n = 95), healthy subjects. Serum levels of copper and zinc were measured by atomic absorption spectrophotometry. The results showed that the serum levels of copper (mg/dL) in patients with digestive cancer (91.6 +/- 27.3, p < 0.05) were significantly higher than in patients with benign digestive disease (75.8 +/- 19.8) or healthy subjects (54.4 +/- 8.9) and the serum levels of zinc (mg/dl) were significantly lower (68.7 +/- 21.9, p < 0.05) compared to benign digestive disease patients (80.1 +/- 18.7) or healthy subjects (100 +/- 11.4 mg/dl). The Cu/Zn ratio was also significantly higher in patients with digestive cancer (1.45 +/- .58, p < 0.05) than those with benign digestive disease (0.95 +/- 0.28) or healthy subjects (0.55 +/- 0.13). Considering a cutoff value of 0.87, the sensitivity of the copper/zinc ratio was 82.2%, with a specificity of 65.7%, a positive predictive value of 45.8% and a negative predictive value of 91.3%. In conclusion, Cu/Zn ratio was found to be considerably higher in patients with digestive cancer compared to age- and gender matched controls, with a sensitivity of 82.2% that might be useful in the evaluation of suspected malignancy. PMID- 9204620 TI - Human papilloma virus 16-18 infection and cervical cancer in Mexico: a case control study. AB - Cervical cancer (CC) is one of the principal public health problems in Mexico. The national mortality rate due to CC was estimated at 21.8 per 100,000 among women over 15 years old during 1994. Despite this high incidence little is known in Mexico about the risk factors for CC. The objectives of the study were to evaluate the association between CC and HPV types 16 and 18 in women living in Mexico City. From August, 1990 to December, 1992, a case-control study was carried out in the metropolitan area of Mexico City. HPV 16-18 types were determined in a sample of 148 CC cases and 204 controls randomly selected from a sample frame representative of the metropolitan area of Mexico City. Sixty cases corresponded to in situ CC and 88 cases to the invasive phase. Determination of HPV 16 and 18 types was done by polymerase chain reaction using primers specific to E6/E7. Results showed that 48.3% of in situ CC cases and 48.8% of invasive CC cases were positive for HPV 16 while only 13.2% were positive among the 204 controls. Association between HPV 16 infection in the in situ cancer cases had an estimated odds ratio (OR) of 5.17 (95% CI 2.60-10.51). In the invasive cervical cancer cases, association between HPV 16 infection and invasive CC in this sample had an OR of 3.84 (95% CI 2.04-7.22). For the total sample, the estimated OR was 5.48 (95% CI 3.07-9.62). In the total sample, those women with a strong positive reaction to PCR were associated with a large increase in the risk, OR of 38.0 (95% CI 8.66-167.1). The prevalence the HPV 18 was 6.7%, only observed in the invasive cervical cancer cases. At present there is general consensus that HPV is the principal causal agent in C C etiology. This study intends to contribute to the knowledge concerning the etiology of cervical cancer. However, it is necessary to consider that the single most effective tool in the reduction of mortality due to cervical cancer has been the Pap test. Secondary prevention has proven to be highly effective in other populations, and this should be viewed as a priority activity for all at-risk populations. Although a vaccine for HPV may be available in the near future its efficacy in primary prevention has yet to be demonstrated. PMID- 9204621 TI - Nutritional state alterations in children with acute lymphoblastic leukemia during induction and consolidation of chemotherapy. AB - The objective of the study was to determine if children with high risk acute lymphoblastic leukemia (ALL) exhibit higher frequency of alterations in nutritional state during the phases of induction and consolidation of chemotherapy than children with low risk ALL, based on the arm muscle area. The design was concurrent comparative cohorts. It was performed at pediatric hematology service of the Hospital General del Centro Medico Nacional "La Raza" and hematology service of the Hospital de Pediatria del Centro Medico Nacional "Siglo XXI". One hundred-five patients were incorporated into the study: 53 with high risk (HR) ALL and 52 with low risk (LR) ALL. Basal measurements of arm circumference and tricipital skinfold were surveyed monthly (for 3 months) by standardized personnel. Altered nutritional state during follow-up was defined as the loss of 10% or more of the arm muscular area (AMA) measured at diagnosis. Statistics of proportion analysis with a significance level of 0.05 and relative risk (RR) with confidence intervals (CI) were calculated. In the first month the RR was 0.77 (CI 0.31-1.87); the LR group was the most affected. In the second month the RR was 7.31 (CI 1.41-38.03); the most affected group was the HR. In the third month the RR was 1.77 (CI 0.60-4.92); the HR group was the most affected. High-risk patients show a higher frequency of nutritional state alterations reflected in AMA during the second month after diagnosis. This may be caused by the more aggressive chemotherapy received by these patients. PMID- 9204622 TI - Prevalence of gestational diabetes in a group of women receiving treatment at the Mexican Institute of Social Security in Aguascalientes, Mexico. AB - The objective of this work was to determine the prevalence and associated clinical variables of gestational diabetes in a group of pregnant women, using a prospective, longitudinal and comparative study. The setting where the study was performed was an urban General Hospital, and outpatient clinics of the Instituto Mexicano del Seguro Social in Aguascalientes City, Mexico. The subjects were 187 pregnant women receiving prenatal care in two health care outpatient clinics where they had given informed consent for a 1-year period. All selected women without a history of diabetes mellitus were studied and scheduled for a full oral glucose tolerance test (OGTT) performed at 24-28 weeks of gestation. Gestational diabetes was diagnosed according to the American Diabetes Association. Results are shown for comparative purposes in three groups: 167 women with normal OGTT, 7 women with one OGTT abnormal value, and 13 women with OGTT criteria for gestational diabetes. The study protocol was approved by the Institutional Review Board. We found a prevalence of 6.9% of gestational diabetes in our study group, and significant differences (p < 0.05) among parity, fasting blood glucose, macrosomy, family history of diabetes, obesity of 90 kg or more, and age > 35 years. Body mass index mean was over 25 kg/m2 in all groups. The 6.9% prevalence of gestational diabetes we found is higher than data between 3.9 and 6% previously reported in Mexico. This could reflect a selection bias of our sample; however, it represents a serious public health problem. Appropriate screening, diagnosis, monitoring, and treatment must be implemented. PMID- 9204623 TI - beta-Lactamase bioassay: a simplified method to determine extended-spectrum beta lactamases (ESBL) in enterobacteria. AB - With the simultaneous use of an isoelectric focusing gel (IEF) and a nitrocefin/cefotaxime bioassay, it is possible to identify the Extended-Spectrum beta-lactamases (ESBL) with precision. A mixture of soft agar and susceptible bacterial cells are layered over the gel following overnight incubation and areas of cell growth are detected where the antibiotic has been hydrolyzed by specific enzymes. This innovative method improves sensitivity and specificity for the identification of ESBLs in those enterobacteria strains producing more than one beta-lactamase and are resistant to third generation cephalosporines. PMID- 9204624 TI - On the correct determination of reference values for serum antibodies against pigeon serum antigen using a group of healthy blood donors. AB - An enzymatic immunoassay was developed in order to evaluate the statistical distribution of IgG serum antibodies against pooled pigeon sera antigen in 102 healthy blood donors (HBD). A non-normal distribution was obtained as demonstrated by abnormal values of skewness (2.02) and kurtosis (6.50). A cut-off point (0.120) was determined from the mean plus 2 standard deviations of the optical density values obtained in the HBD group. This value was able to segregate 94% of subjects. However, when calculation of the mean less 2 SD was performed to delimit 95% of the samples, an aberrant negative value was obtained. In contrast, when the nonparametric method of percentile calculation was applied, an optical density value of 0.130 discriminated 97.5% of samples. In addition, the interval between p97.5 and p2.5 delimited 95% of samples. We conclude that when reference values and cut-off point are determined from an enzymatic immunoassay, careful analysis of the statistical distribution of reference values is necessary in order to avoid the inappropriate application of parametric procedures as demonstrated in this study for antibodies against pigeon serum antigens. PMID- 9204625 TI - Chronic myelocytic leukemia in accelerated phase with i(17) (q10) and loss of p53 gene. Case report. AB - Chronic myelogenous leukemia (CML) is a clonal disorder that presents with a stable period followed by an accelerated phase. The most frequent chromosomal abnormality described is t(9;22). Alterations of chromosome 17, where p53 is located, have been described during transformation. We studied a 23-year-old male who presented with chronic myelogenous leukemia. The karyotype demonstrated 46,XY,t(9;22) (q34;q11) in 12% of mitoses and hyperdiploidy in 43%. Forty-six months later the patient suffered a blast crisis characterized by absolute basophilia; the cytogenetic study demonstrated 48,XY,+8,t(9;22) (q34;q11), +der(22)t (9;22) (q34;q11), +i(17)(q10) in 18% of the mitoses, 46,XY, t(9;22) (q34;q11) in 34% and hyperdiploidy in 23%. Since there was i(17)(q10) during this stage, a retrospective DNA study of the biopsy material before and after the transformation was performed. In the chronic phase, p53 was present in normal amounts, during transformation there was loss of genetic material from the p53 region. The protein product of suppressor gene p53 normally works holding the proliferation of cells. When there is the formation of an isochromosome, genetic material is lost; thus, in this patient, p53 was deleted upon the observation of i(17). Lastly, this case shows how DNA can be extracted from slides; this technique is novel and can be used for retrospective studies when paraffin blocks or fresh tissue are not available. PMID- 9204626 TI - Polygraphic characterization of the sleep-epilepsy patterns in a hydranencephalic child with severe generalized seizures of the Lennox-Gastaut syndrome. AB - This is the report of a hydranencephalic child with severe generalized seizures of the Lennox-Gastaut Syndrome (LGS) who lacked the development of the entire cerebral hemispheres and had preserved the brain stem, cerebellum, hypothalamus and a portion of the thalamus as evidenced by radiological and/or physiological studies. Conventional polygraphic sleep studies in these patients showed presence of scalp EEG and other peripheral, somatic and vegetative signs characterizing the wakefulness, quiet sleep and active sleep stages. Absence of the vertex waves and disrupted sleep spindles were the major qualitative EEG abnormalities. In contrast, quantitative abnormalities in duration, latency and number of sleep cycles found in this patient were similar to those found in other children with Idiopathic Lennox-Gastaut Syndrome (ILGS). A substantial reduction in the number of interictal EEG spikes and a shortening of the ictal clonic EEG activities without concomitant EMG jerks were the most distinct epileptiform abnormalities in this child. In contrast, his basic polygraphic patterns of the tonic and apneic seizures were similar to those found in other children with ILGS. Data obtained from this child suggest that both the sleep stages and the generalized seizures of the ILGS basically depend more on the integrity of the brain stem than on the telencephalic structures. PMID- 9204627 TI - Primary hyperparathyroidism due to parathyroid carcinoma. AB - Most cases of primary hyperparathyroidism are due to either a parathyroid adenoma or to parathyroid hyperplasia. Parathyroid carcinoma is a very rare cause of hyperparathyroidism. Although the diagnosis of parathyroid carcinoma is usually established based on pathological criteria of vascular and capsular invasion, some clinical and biochemical features differentiate it from benign forms of hyperparathyroidism. We report the case of a middle-aged woman with a long standing history of nephrolithiasis, who presented with a palpable neck mass, weight loss, severe hypercalcemia and hypophosphatemia, as well as very high serum levels of intact parathyroid hormone. Surgical neck exploration revealed a large tumor that invaded trachea, esophagus, reccurrent laryngeal nerve, right apical pleura and right carotid artery. Pathological examination confirmed the invasive nature of the tumor. Along with the case report, we review the literature and discuss the diagnostic and therapeutic options of this rare condition. PMID- 9204628 TI - Scientific contributions of Mexican scientists. PMID- 9204629 TI - Thinner abusers. PMID- 9204630 TI - Local institutional development and relief in Ethiopia: a kire-based seed distribution programme in North Wollo. AB - Highlighted here is the important role played by community-based organisations in relief supply operations. In the context of an emergency seed supply project in northern Ethiopia in 1995, it examines the participation of burial societies, known as kires, in targeting, distribution and management. The paper illustrates that factors of local institutional legitimacy, transparency and accountability are central, both to the effective representation of community views and to long term partnerships between local institutions and non-governmental organisations. PMID- 9204631 TI - Estimating the capacity of warehouses. AB - The required capacity of warehouses for use in humanitarian emergencies is very often overestimated. This paper reviews some of the principal factors affecting warehouse capacity for emergency humanitarian operations in developing countries. The growing difference between modern commercial warehousing practice and the approach typically used in humanitarian emergencies is highlighted. The principal constraints on capacity are identified and discussed. A simple method for estimating the storage capacity of warehouses for emergency operations is then presented. PMID- 9204632 TI - Research in the war zones of Eritrea and northern Ethiopia. AB - The paper engages in the discussion of conducting research in war zones, initiated in Disasters by Barakat and Ellis. It looks specifically at possibilities for research in the war zones of Eritrea and Northern Ethiopia during the 1980s, and notes the ways in which this context differs from wars in the former Yugoslavia to which Barakat and Ellis mainly refer. The authors suggest that the unique context of every internal war, and the institutional actors that converge around this context, create both the potential and the demand for particular kinds of information, especially when humanitarian programmes involving international donors are under way. De-contexualising research in war zones from the specific context in which it occurs, in order to derive general guidelines, can thus be problematic. PMID- 9204633 TI - Evaluation of disaster assistance. PMID- 9204634 TI - Context and consistency: the Canadian connection. AB - Issues related to context effects in hypnosis research are briefly reviewed. The contributions of Canadian hypnosis researchers to current theory and research on context effects are acknowledged. Bowers and colleagues at the University of Waterloo emphasized the scope and subtlety of contextual influences on correlates of hypnotic suggestibility, and they promoted the development of a consistency motivation theory of context effects. Spanos and colleagues at Carleton University generalized context effects within the domain of hypnosis, prompting extension of this work to general personality measurement. Implications of findings on consistency motivation for hypnosis research are discussed in terms of person-by-situation interactions. PMID- 9204635 TI - Suggestibility or hypnosis: what do our scales really measure? AB - Conceptually, hypnotizability has always been defined as the increase in suggestibility produced by hypnosis. In practice, hypnotizability is measured as suggestibility following a hypnotic induction. The data indicate that these are different constructs. Although the induction of hypnosis increases suggestibility to a substantial degree, the correlation between hypnotic and nonhypnotic suggestibility approximates the reliability coefficients of so-called hypnotizability scales. This indicates that hypnotic susceptibility scales are better measures of waking suggestibility than they are of hypnotizability. The discordance between conceptual and operational definitions of hypnotizability can be resolved either by changing the conceptual definitions of hypnosis and hypnotizability or by reinterpreting hypnotizability scores as indexes of nonhypnotic, imaginative suggestibility. PMID- 9204636 TI - Have the hypnotic susceptibility scales outlived their usefulness? AB - Hypnosis experiments often involve preselecting high- and low-scoring participants on the basis of one or more hypnotic suggestibility scales, and then studying the differences between these two groups. A number of possible critiques of this method are entertained in this article. For example, sociocognitive theorists would seem better advised to directly manipulate the variable or variables they believe underlie the differences in hypnotic susceptibility, and neodissociative theorists would seem well advised to question whether the scales muddle important distinctions in underlying mechanisms. In addition, parallels are drawn with developments in other areas of research, such as intelligence. PMID- 9204637 TI - Automaticity and hypnosis: a sociocognitive account. AB - This article provides an overview of a new theory of suggested involuntariness in hypnosis, developed in conjunction with Irving Kirsch. The theory is based on the following ideas. First, high hypnotizable participants enter hypnosis with a conscious intention to feel and behave in line with suggested experiences and movements. Second, people who are easily hypnotized hold firm expectations that they will succeed in following the suggestions of the hypnotist. Third, the intention and expectation in turn function as response sets in the sense that they trigger the hypnotic response automatically. Fourth, given the intention to feel and behave in line with the hypnotist's suggestions, hypnotized individuals show no hesitation to experience the suggested movements as involuntary because (a) these movements are actually triggered automatically, and (b) the intention to cooperate with the hypnotist as well as the expectation to be able to do so create a heightened readiness to experience these actions as involuntary. PMID- 9204639 TI - Admissibility and per se exclusion of hypnotically elicited recall in American courts of law. AB - State v. Mack (1980) ruled that hypnotically elicited testimony is per se excluded from Minnesota law courts; this court also ruled that police could employ hypnosis in an attempt to construct an independently corroborated case. In recent years, there have been moves to rescind this exclusion; this raises a question of the probative value of such additional information when it is uncorroborated. This situation is compared with that of the polygraph as an index of deception: Like hypnosis, it is excluded per se in most American jurisdictions. Some legal decisions in Wisconsin are used to illustrate one alternative to the per se exclusion approach. Admissibility of scientific evidence in American courts of law has been based on a criterion of "general acceptability within the relevant scientific community," as first elucidated in Frye v. United States (1923). Recently, the U.S. Supreme Court overturned the Frye decision in Daubert v. Merrell Dow Pharmaceuticals, Inc. (1993), by making general acceptability but one of several admissibility criteria. Three Daubert based decisions, one involving hypnosis and all concerned with "recovered repressed memories," indicate some problems in law posed by Daubert. PMID- 9204638 TI - The state of the "state" debate in hypnosis: a view from the cognitive-behavioral perspective. AB - For most of the past 50 years, hypnosis research has been driven by a debate about whether hypnotic phenomena can be best described and understood as the product of an altered state of consciousness. The meanings of some of the pivotal concepts in this debate and the nature of the phenomena that gave rise to them were ambiguous at the outset and led to misconceptions and surplus meanings that have obscured the debate through most of its history. The nature of the posited hypnotic state and its assumed consequences have changed during this period, reflecting the abandonment of untenable versions of hypnotic state theory. Carefully conducted studies in laboratories around the world have refined our understanding of hypnotic phenomena and helped identify the critical variables that interact to elicit them. With the maturation of the cognitive-behavioral perspective and the growing refinement of state conceptions of hypnosis, questions arise whether the state debate is still the axis about which hypnosis research and theory pivots. Although heuristic value of this debate has been enormous, we must guard against the cognitive constraints of our own metaphors and conceptual frameworks. PMID- 9204640 TI - Hypnotic theorizing: spring cleaning is long overdue. AB - Since the beginnings of animal magnetism and hypnosis, clinical and experimental theories have developed mostly in isolation. On the clinical side, armchair theorizing, based on clinicians' observations and interpretations of their clients' narratives, have laid the foundations of most theories, past and present. On the experimental side, attachment to specific theories has guided researchers in their choice of methodologies and designs. What seemed important was to show how correct one's preferred view was or how incorrect the "other" could be. This unfortunate one-sided perception led to an experimental stalemate where most experiments could be interpreted from any point of view. If the tendency to theorize on what one sees, hears, and believes must come to an end, the practice of performing truncated experiments dedicated to the glory of one's preferred theory must also be relegated to the past. And while we are at it, whether clinical or experimental, the use of concepts or constructs that cannot be clearly operationalized should be dismissed to avoid the type of serious social consequences the hypnotic community has been facing in the past decades. PMID- 9204641 TI - Why scientific hypnosis needs psychoanalysis (or something like it). AB - The author contends that some contemporary hypnosis theories are restricted and narrow in scope, rendering them unnecessarily isolated from mainstream models of human development, psychopathology, and personality functioning. They seem to explain hypnosis and little else. The author contrasts this with psychoanalysis, which, although sometimes overly expansive, does nonetheless lend itself to the generation of specific hypothesis via careful deduction from a general theory of human behavior and experience. For illustrative purposes, the author criticizes the sociocognitive perspective of hypnosis contending that at present it is too narrowly inductive in focus, overvalues social influence, and has its own problems with reification. The author suggests remedies for these difficulties. PMID- 9204642 TI - EEG concomitants of hypnotic susceptibility. AB - Numerous historical attempts have been directed at understanding electrocortical concomitants of hypnosis. Today, with the availability of more sophisticated multichannel recording technologies and signal-processing approaches, it is possible to reconsider and update previous attempts. The most solid relationship between electrocortical activity and hypnotizability exists in the EEG theta frequency range. Given the stable electrocortical differences found in high and low susceptible individuals, the question arises whether we can use additional EEG measures to help understand the nature of these individual differences. One possible alternative is the pointwise or fractal dimension, which we examined during baseline conditions with high and low hypnotic susceptible individuals. The dimensionality measures suggest that high susceptible individuals display underlying brain patterns associated with imagery, whereas low susceptible individuals show patterns consistent with cognitive activity (i.e., mental math). This type of speculation is similar to that of Tellegen, who makes a distinction between imaginative versus realistic responding. Future research should address the exact nature of the underlying process (imagination, effortlessness, suggestibility, etc.) seen in high and low susceptible individuals. PMID- 9204643 TI - What this field needs is a good nomological network. AB - Research in the field of hypnosis lacks a coherent structure on which to build. This lack of a mature nomological network stems from fundamental disagreements concerning the construct validity of hypnotizability, which in turn stem in part from different research practices across laboratories. For these reasons, the field has had less impact on psychology and medicine than is warranted by the numerous sophisticated scientific studies that have been conducted during the past three decades. PMID- 9204644 TI - Convergence in understanding hypnosis? Perhaps, but perhaps not quite so fast. AB - The study of hypnosis has been plagued by conflict. Although a more recent trend has been the search for convergence among disparate points of view, two highly salient issues remain contentious: the question of whether hypnosis involves alterations in consciousness, and the nature and correlates of individual differences in hypnotic response. Theoretical convergence is a laudable goal, but not at the expense of obscuring the complexity of hypnosis as a state of altered consciousness, a cognitive skill, and a social interaction. Perhaps the best prescription for convergence in hypnosis is the cautious conviction advocated by Kenneth S. Bowers and so clearly exemplified in his own research. PMID- 9204645 TI - Nifedipine for hiccups. PMID- 9204646 TI - Re: Use of rectal medications in palliative care. PMID- 9204647 TI - Re: Use of rectal medications in palliative care. PMID- 9204648 TI - Ketorolac: continuous subcutaneous infusion for cancer pain. PMID- 9204649 TI - Alkalinization is troublesome in advanced cancer patients with dyspnea. PMID- 9204650 TI - Blood transfusions in patients with advanced cancer. PMID- 9204651 TI - A randomized, controlled trial of intravenous clodronate in patients with metastatic bone disease and pain. AB - To evaluate the effectiveness of intravenous clodronate in ameliorating refractory bone pain in patients with metastatic bone disease, 60 patients with established osseous metastases and persistent bone pain were randomized to receive either clodronate (600 mg or 1500 mg in 500 mL of normal saline) or 500 mL of saline as placebo. After 2 weeks, the patients were crossed over to receive the alternate treatment. After another 2 weeks, each patient and investigator made a blinded choice. Daily visual analogue scales (VAS) and analgesic diaries were recorded throughout the study period. Forty-six patients were evaluable (77%). A treatment x period interaction was identified in the VAS and daily morphine equivalent dose (DMED) scores. First period analysis of the VAS scores for general pain, pain at rest, and pain upon movement demonstrated an average reduction of 13, 14, and 24 mm, respectively, from baseline, but were not significantly different from changes following placebo. The average change in DMED was -6.4 (SE = 2.9) following clodronate and was +24.6 (SE = 14.9) following placebo (p = 0.03). In the blinded choice of which agent resulted in improvement in pain, 26 (57%) patients chose clodronate, 12 (26%) chose placebo, and eight (17%) had no preference (p = 0.0021). For the investigators who also made a blinded selection, clodronate was chosen in 30 (65%) patients, placebo in ten (22%) patients, and no difference was apparent in six (13%) (p < 0.0001). Intravenous clodronate appeared to have analgesic effect in patients with refractory bone pain due to metastatic bone disease. The optimal dose and duration of effect require further evaluation, particularly in patients with stable disease and persistent bone pain. PMID- 9204652 TI - The effects of pre-emptive treatment of postherpetic neuralgia with amitriptyline: a randomized, double-blind, placebo-controlled trial. AB - Seventy-two patients older than 60 years of age who received a diagnosis of herpes zoster (HZ) were entered into a randomized, double-blind, placebo controlled trial of daily amitriptyline 25 mg. Treatment with either amitriptyline or placebo continued for 90 days after diagnosis. Pain prevalence at 6 months was the primary outcome. Results showed that early treatment with low dose amitriptyline reduced pain prevalence by more than one-half (p < 0.05; odds ratio, 2.9:1) This finding makes a strong case for the pre-emptive administration of amitriptyline, in combination with an antiviral drug, to elderly patients with acute herpes zoster. PMID- 9204653 TI - Vasomotor rhinitis and sphenopalatine ganglion block. AB - The effectiveness of the sphenopalatine ganglion (SPG) block for the relief of symptoms in chronic vasomotor rhinitis was assessed in 30 patients of both genders. The number of blocks required for complete relief was three (range from two to four) at weekly intervals in 66.7% of volunteers. There was no recurrence of symptoms during a follow-up period of 12-20 months in 29 patients, and one patient was symptom free for 8 months. The technique is simple and can be performed as an outpatient procedure without side effects. PMID- 9204654 TI - The management of diabetes in patients with advanced cancer. AB - The management of diabetes mellitus is often complicated in patients with advanced cancer. Anorexia and nausea or vomiting make caloric intake erratic. The use of diabetogenic medications such as glucocorticoids can produce profound hyperglycemia. Many malignant tumors cause derangement in intermediary metabolism and abnormal glucose tolerance in up to one-third of patients. Both hyperglycemia and hypoglycemia impair the quality of life of dying patients. Swings in blood sugar should be avoided wherever possible, but aggressive blood sugar monitoring also impairs quality of life. This paper discusses issues in the management of diabetes in patients with advanced cancer and suggests guidelines for maintaining glycemic control without excessive interventions. PMID- 9204656 TI - Octreotide may prevent definitive intestinal obstruction. AB - Partial bowel obstruction is indistinguishable from definitive obstruction at the onset of symptoms. No consensus exists regarding the treatment of potentially reversible states of bowel obstruction. On the basis of previous experience, octreotide was used in two patients with chronic intestinal obstruction, resulting in good control of intestinal symptoms and maintenance of a prolonged adequate intestinal transit, preventing the occurrence of definitive bowel obstruction. The results observed stress the early use of octreotide in such delicate clinical situations. PMID- 9204655 TI - Is a "palliative" patient always a palliative patient? Two case studies. AB - Review of the literature suggests that misdiagnosis of terminal illness is infrequent. In the first 6 months of the recently established Edmonton Regional Palliative Care Program, two of 330 referrals proved to be in the category of erroneous diagnosis of terminal disease. These two cases are reported, along with discussion of aspects of the time-honored usefulness of careful history and physical examination. This experience highlights the importance of assessment, investigation, and aggressive therapy, even in "terminal" patients, including those in the geriatric population. PMID- 9204657 TI - Severe opioid toxicity and somatization of psychosocial distress in a cancer patient with a background of chemical dependence. AB - A case of severe opioid toxicity is described in a 52-year-old cancer patient. The patient presented with classical clinical features of central hyperexcitability associated with opioid toxicity: delirium, myoclonus, hallucinations, hyperalgesia, and a possible seizure. This patient had a background of severe psychosocial distress and somatization in addition to a history of benzodiazepine dependence and alcohol abuse. The occurrence of opioid toxicity in this patient highlights the risks of a unidimensional approach to cancer pain, which ignores the non-organic components of pain, such as psychosocial distress, which will not respond to escalating doses of opioid medication. PMID- 9204658 TI - Increased phantom limb pain as an initial symptom of spinal-neoplasia. AB - Phantom limb pain is a common sequela of amputation. Studies suggest that over time, there is a decrease in frequency and intensity of phantom pain. Persistently increased phantom pain has been seen in benign lesions affecting the peripheral and central nervous system. We present a 74-year-old woman who had a left above-knee amputation for leiomyosarcoma of the foot 24 years previously. She had been free of disease and ambulated independently until 1 month before hospitalization, when she noted increasing pain in her phantom foot. At the time of admission, she had developed increasing low back pain and was diagnosed with adenocarcinoma of unknown primary. Work-up confirmed involvement of the L4 vertebral body with epidural and paraspinal disease. We believe this is the first reported case of worsening phantom limb pain resulting from a spinal metastasis. We review the literature on the potential implications of increased phantom pain. PMID- 9204659 TI - TD from risperidone? PMID- 9204660 TI - SSRI for body dysmorphic disorder. PMID- 9204661 TI - Valproic acid for Kleine-Levin syndrome. PMID- 9204662 TI - Catecholamines in ADHD: a postscript. PMID- 9204663 TI - Catatonia in children and adolescents. PMID- 9204664 TI - Gender identity disorder: a review of the past 10 years. AB - OBJECTIVE: To review the clinically relevant literature on gender identity disorder (GID) in children and adolescents over the past 10 years. METHOD: All literature referring to gender identity and children or adolescents from 1985 on was reviewed. RESULTS: Changes in the DSM-IV nomenclature include (1) adoption of the single diagnosis of GID to apply to children, adolescents, and adults; (2) changes in the format of the criteria; and (3) placement in the section "Sexual and Gender Identity Disorders." Rates of associated psychopathology in children with GID are comparable with those in children with other psychiatric disorders, particularly disorders that are internalizing in form. Biological and psychosocial factors thought to be relevant in the development of GID are reviewed. CONCLUSIONS: Research is required to elucidate the complicated interaction between biological and psychosocial factors in the development of GID and to evaluate treatment efficacy. PMID- 9204665 TI - Psychosocial functioning of homeless children. AB - OBJECTIVE: To investigate the psychosocial characteristics of homeless children and their parents. METHOD: Homeless families were assessed within 2 weeks of admission to seven hostels and were compared with a group of housed families matched for socioeconomic status. Measures included a semistructured interview, the General Health Questionnaire (GHQ), the interview Schedule for Social Interaction, the Child Behavior Checklist (CBCL), the Communication domain of the Vineland Adaptive Behavior Scales, and height and weight percentiles. The sample consisted of 113 homeless families (249 children aged 2 through 16 years) and 29 comparison families (83 children). RESULTS: Homeless families primarily consisted of single mothers and an average of two children, who had become homeless because of domestic violence (56%) or violence from neighbors (29%). Homeless mothers reported high rates of previous abuse (45%) and current psychiatric morbidity (49% caseness on the GHQ) and poor social support networks compared with housed controls. Homeless children were more likely to have histories of abuse, living in care, and being on the at-risk child protection register and less likely to have attended school or a preschool/day-care center since admission to the hostel. They also had delayed communication and higher CBCL scores. Maternal GHQ scores best predicted CBCL caseness. CONCLUSIONS: Homeless mothers and children have high rates of psychosocial morbidity, which are related to multiple risk factors and chronic adversities. Their complex needs should be best met by specialized and coordinated health, social, and educational services. PMID- 9204667 TI - Factors associated with child mental health service use in the community. AB - OBJECTIVE: To determine the association of parent, family, and child factors with mental health services need and utilization. METHOD: Possible determinants of services need and utilization were assessed in a general population sample of 2,227 children aged 4 to 18 years. RESULTS: 3.5% of the total sample had been referred for mental health services within the past year. The most potent factors associated with service need and utilization were the child's problem behaviors (both internalizing and externalizing) and academic problems and family stress. Socioeconomic factors and the child's sex were not in itself associated with help seeking factors. Parental psychopathology, life events, and family psychopathology lowered the parents' threshold for evaluating the child's behavior as problematic but did not increase the likelihood of referral. CONCLUSION: Referred children are more likely to live in families under stress than are children with the same level of problems who live in well-functioning families. Clinicians and researchers who make inferences from findings in clinical samples should realize, therefore, that children from problem families are overrepresented in their samples. PMID- 9204666 TI - Mental disorders and service utilization among youths from homeless and low income housed families. AB - OBJECTIVE: To assess the mental health of homeless and poor housed youths, using the National Institute of Mental Health (NIMH) Diagnostic Interview Schedule for Children (DISC) Version 2.3, and to examine mental health service use. METHOD: As part of a comprehensive study of homeless and housed families Worcester, MA, data were collected on 41 homeless and 53 poor housed (never homeless) youths aged 9 to 17 using both the parent and youth versions of the DISC. RESULTS: On the basis of the parent version of the DISC, current (6-month) prevalence rates of DSM-III R disruptive behavior, affective, and anxiety disorders were comparable in homeless and housed youths but higher than rates found among youths in the NIMH sponsored Methods for the Epidemiology of Child and Adolescent Mental Disorders (MECA) Study, which used the same diagnostic measure. Approximately 32% of the combined sample of homeless and housed youths had a current mental disorder accompanied by impairment in functioning. Mental health service use in the preceding 6 months among youths who had one or more current disorders and associated impairment ranged from 20% to 35%. A subgroup of youths with one or more current disorders and poor global functioning had never received treatment. CONCLUSIONS: This sample of homeless and housed youths was found to have high rates of current mental disorders. Use of mental health services by children with mental health needs was low, particularly for youths with poor overall functioning. PMID- 9204668 TI - Expressed emotion toward children with behavioral inhibition: associations with maternal anxiety disorder. AB - OBJECTIVE: To examine the role of maternal psychopathology in influencing "expressed emotion" (EE) directed toward children with behavioral inhibition (BI) or psychiatric disorders. METHOD: Maternal EE was assessed via Five-Minute-Speech Sample in two samples of children previously evaluated for child and maternal lifetime prevalence of DSM-III disorders and assessed via laboratory observations for BI. The authors previously reported that maternal EE was associated with BI and with the number of child behavior and mood disorders in these samples. The at risk sample (N = 30) consisted of mothers with panic disorder and psychiatric controls and their 4-through 10-year-old children. The Kagan sample (N = 41) consisted of children selected at age 21 months as BI or uninhibited and followed through age 11. RESULTS: Interaction effects were found: in mothers with anxiety disorders, but not those without, maternal criticism (a component of EE) was significantly associated with child BI, independently of the child's number of disorders. Similarly, in mothers with anxiety disorders only, maternal criticism was significantly associated with a high number of child disorders. CONCLUSIONS: The relationships between mothers who have anxiety disorders and their children who have BI or psychiatric disorders may be marked by criticism or dissatisfaction. If confirmed, these findings offer opportunities for appropriate interventions. PMID- 9204669 TI - At risk for anxiety: I. Psychopathology in the offspring of anxious parents. AB - OBJECTIVE AND METHOD: Children of parents with anxiety disorders, depressive disorders, mixed anxiety/depressive disorders, and no psychiatric disorder were assessed with semistructured interviews to determine rates of overall psychopathology and to determine specifically the presence of anxiety disorders. RESULTS: Children of the three "high-risk" groups were significantly more likely to have a diagnosable disorder (including anxiety disorders) than offspring of normal parents, but there were no differences among the children from the three parental diagnostic groups. However, when examined specifically for anxiety disorders, offspring of anxious parents were significantly more likely to have only anxiety disorders. Offspring of depressed or mixed anxious/depressed parents had a broader range of disorders and more comorbid disorders. Family socioeconomic status was related to the probability that a child would have a disorder. CONCLUSIONS: Anxiety disorders are common among offspring of anxious and depressed parents. However, when a parent has depression, children exhibit a broader range of psychopathology than when a parent has an anxiety disorder alone. PMID- 9204670 TI - Startle modulation in children at risk for anxiety disorders and/or alcoholism. AB - OBJECTIVE: To examine the startle reflex as a possible vulnerability marker among offspring of parents with anxiety disorders and/or alcoholism. METHOD: The subjects were 66 male and female offspring (aged 10 to 17 years) of proband who participated in a family study of comorbidity of alcoholism and anxiety disorders. Testing consisted of examining the startle reflex and its modulation by prepulse stimuli (prepulse facilitation and prepulse inhibition). RESULTS: Different components of the startle discriminated among children of parents with anxiety disorders, children of alcoholics, and children of normal controls. Specifically startle magnitude was elevated in children with a parental history of an anxiety disorder, whereas startle habituation and prepulse inhibition were impaired in children with a parental history of alcoholism. CONCLUSION: These findings suggest that individual differences in the startle reflex may serve as a vulnerability marker for the development of anxiety disorders and alcohol problems. PMID- 9204671 TI - Relationship of perceived competencies, perceived social support, and gender to substance use in young adolescents. AB - OBJECTIVE: This survey study explores the relationship between area-specific perceived self-competence, perceived social support, gender, and substance use in young adolescents. METHOD: Questionnaires were administered to 140 male and 131 female adolescents attending middle school to assess self-perception of competencies, social support, and substance use. Correlations were performed between the predictor variables and the substance use measures. Hierarchical multiple regressions were also used to identify potential interactions between gender, perceived competencies, and perceived social support in the prediction of specific substances. RESULTS: Higher perceived scholastic competence was associated with less substance use in both genders. In boys, more perceived support from teachers, and to a lesser degree parents, was associated with less substance use, particularly in those with low scholastic competence. In girls, social support was unrelated to substance use except for support from classmates, which was associated with more cigarette and marijuana use. However, in girls with low scholastic competence, more support from peers was consistently associated with more substance use. CONCLUSIONS: The gender differences in risk factors for early substance use identified in this study deserve further investigation, in view of their potential relevance for adolescent substance abuse prevention and early intervention. PMID- 9204672 TI - Clinical characteristics of psychiatrically referred adolescent outpatients with substance use disorder. AB - OBJECTIVE: There is increasing interest in the developmental relationship of psychopathology and substance use disorders (SUD) in youths. Because the bulk of literature is focused in inpatient or incarcerated youths, inferences and generalizations are limited in outpatient settings. The clinical characteristics of psychiatrically referred outpatients were studied to determine whether differences existed in the nature and severity of comorbid psychiatric disorders when substance abuse was involved. METHOD: All diagnoses were derived from structured psychiatric interviews completed on all youths on intake assessment. Adolescents with an identified SUD (n = 38) were compared with those without SUD (n = 321) on a number of variables including past and current psychopathology, cognitive and school functioning, and overall impairment. RESULTS: Eleven percent of referred outpatients (mean age = 15.9 +/- 1.3 years) met full criteria for a SUD by parental report. Controlling for age, adolescents with SUD had higher risk for mood and disruptive behavioral disorders compared with psychiatric controls. In the majority of cases, the onset of psychopathology preceded the onset of SUD by at least 1 year. The group with SUD also had lower overall functioning and more school dysfunction and psychiatric hospitalizations than their non-SUD peers. CONCLUSIONS: Despite the small number of adolescents with SUD in this sample, these data indicate that SUD is common in outpatient psychiatry referrals. Youths with SUD appear to be at increased risk for more psychopathology and dysfunction in a number of domains. PMID- 9204673 TI - How suggestible are preschool children? Cognitive and social factors. AB - OBJECTIVES: In this series of studies, the authors sought to determine some of the cognitive and social boundary conditions that can undermine the accuracy of young children's reporting. Care was taken to include events and interviewing variables that more accurately reflect the experiences of children in real-world investigations of alleged sexual abuse. Videotaped interviews with preschool children were presented to experts to determine how adept they are at distinguishing between true and false accounts. METHOD: All the studies were designed to investigate the susceptibility to suggestion in young preschool children. The difference between studies was the form of that suggestion and the nature of the event to which the children were exposed. All studies measured recall accuracy, false assent rate, and the change in these outcomes over time and/or successive interviews. RESULTS: Very young preschool children (aged 3 and 4 years) were significantly more vulnerable to suggestions than were older preschool children (aged 5 and 6 years). The number of interviews and the length of the interval over which they were presented resulted in the greatest level of suggestibility. CONCLUSIONS: While some types of events (negative, genital, salient) were more difficult to implant in children's statements, some children appeared to internalize the false suggestions and resisted debriefing. These children's false statements were quite convincing to professionals, who were unable to distinguish between true and false accounts. PMID- 9204674 TI - Relationship between sexual abuse, gender, and sexually inappropriate behaviors in seriously mentally ill youths. AB - OBJECTIVE: To examine gender differences in sexual abuse histories and in the development of inappropriate sexual behaviors in a sample of seriously mentally Ill youths. METHOD: A retrospective chart review was completed for all patients from 1987 through 1992 at a tertiary care public sector psychiatric hospital for youths (N = 499). Subjects were categorized by gender, sexual abuse status, and whether they had sexually reactive or victimizing behaviors. RESULTS: Girls were more likely to have been sexually abused, and their abuse histories were more severe. Sexual behavior problems in girls were almost exclusively associated with sexual abuse, whereas 29% of boys with victimizing behaviors had no sexual abuse history. Among sexually abused youths, boys were more likely to display victimizing behaviors, whereas both genders displayed similar rates of sexually reactive behaviors. Of the 19 girls who displayed victimizing behaviors, 95% were chronically sexually abused and one third had also received a major injury due to physical abuse. CONCLUSIONS: Boys appear to have a lower threshold of abuse exposure required to develop sexually inappropriate behaviors and are significantly more likely to display victimizing behaviors. Conversely, victimizing behaviors in girls may require a catastrophic maltreatment history. These gender differences should be incorporated into treatment interventions directed at sexual abuse victims. PMID- 9204675 TI - Case study: allegations of abuse created in a single interview. AB - OBJECTIVE: To illustrate how young children can be induced to make false allegations of sexual abuse. METHOD: The author presents a case that is unusual because elaborate, detailed allegations of sexual abuse came about during a single interview, the interviewer was a baby-sitter rather than a mental health professional, and the interview was recorded on tape. RESULTS: Children can be induced to make elaborate, detailed false statements after being subjected to repetitive, suggestive, and leading questions during a single interview. CONCLUSIONS: Child abuse investigators should determine the origin and evolution of allegations of abuse. Children should not be removed from their parents if is likely that the allegations against the parents are false. PMID- 9204676 TI - Does psychiatric history bias mothers' reports? An application of a new analytic approach. AB - OBJECTIVE: To evaluate whether mothers' psychiatric history biases reports of their children's behavior problems, mothers' and teachers' reports of children's behavior problems were compared using a recently developed statistical approach. METHOD: Child Behavior Checklists and Teacher's Report Forms were completed by mothers and teachers, respectively, about 801 six-year-old children. Mother's history of major depression, anxiety disorders, and substance use disorder was assessed by using the National Institute of Mental Health Diagnostic Interview Schedule. Generalized estimating equations were used for data analysis. RESULTS: According to both teachers and mothers, maternal history of major depression was associated with more internalizing problems; the association was significantly stronger when mothers were the informants. Mothers with history of any psychiatric disorder reported more externalizing problems in their children than expected, whereas teachers' reports of externalizing behaviors were unrelated to maternal psychiatric history. These findings could not be explained by variations in children's behaviors across settings. CONCLUSION: The generalized estimating equation models enabled simultaneous examination of whether children of depressed mothers have excess behavior problems and whether depressed mothers overreport behavior problems in their children. The results indicate that children of depressed mothers have more internalizing problems. In addition, depressed mothers overstate and overgeneralize their offspring's behavior problems. This study broadens the concerns with reporting bias beyond maternal depression to include other psychiatric problems. The results emphasize the potential for bias in family history studies that rely on informants. PMID- 9204677 TI - Schedule for Affective Disorders and Schizophrenia for School-Age Children Present and Lifetime Version (K-SADS-PL): initial reliability and validity data. AB - OBJECTIVE: To describe the psychometric properties of the Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime version (K-SADS-PL) interview, which surveys additional disorders not assessed in prior K SADS, contains improved probes and anchor points, includes diagnosis-specific impairment ratings, generates DSM-III-R and DSM-IV diagnoses, and divides symptoms surveyed into a screening interview and five diagnostic supplements. METHOD: Subjects were 55 psychiatric outpatients and 11 normal controls (aged 7 through 17 years). Both parents and children were used as informants. Concurrent validity of the screen criteria and the K-SADS-PL diagnoses was assessed against standard self-report scales. Interrater (n = 15) and test-retest (n = 20) reliability data were also collected (mean retest interval: 18 days; range: 2 to 36 days). RESULTS: Rating scale data support the concurrent validity of screens and K-SADS-PL diagnoses. Interrater agreement in scoring screens and diagnoses was high (range: 93% to 100%). Test-retest reliability kappa coefficients were in the excellent range for present and/or lifetime diagnoses of major depression, any bipolar, generalized anxiety, conduct, and oppositional defiant disorder (.77 to 1.00) and in the good range for present diagnoses of posttraumatic stress disorder and attention-deficit hyperactivity disorder (.63 to .67). CONCLUSION: Results suggest the K-SADS-PL generates reliable and valid child psychiatric diagnoses. PMID- 9204678 TI - Outcome of multimodal day treatment for children with severe behavior problems: a five-year follow-up. AB - OBJECTIVE: To evaluate the long-term outcome of a multimodal day treatment program for children with severe behavior problems and to identify factors that may predict a positive outcome. METHOD: Thirty-three children who completed a day treatment program were assessed using a prospective, single-cohort design tested at intake, discharge, and 5-year follow-up. The child's functioning was assessed using the Revised Child Behavior Profile (RCBP), Hare Self-Esteem Scale, Depression Self-Rating Scale, Hopelessness Scale for Children, Index of Peer Relations, and a 5-point ordinal scale for scholastic reintegration. RESULTS: Repeated-measures analyses of variance showed that improvement was maintained on all measures between intake and 5-year follow-up. A stepwise multiple regression showed that 92% of the adjusted variance in the behavioral functioning of the children at 5-year follow-up, as assessed by the RCBP, was explained by parental cooperation, initial RCBP total and externalizing scores, and history of problem pregnancy. CONCLUSIONS: Children who were admitted to a day treatment setting appear to function well globally, even 5 years after discharge. Parental cooperation was the most important variable in predicting positive outcome. PMID- 9204680 TI - My father, Frankenstein: a child's view of battering parents. PMID- 9204679 TI - Relationship of depressive, conduct, and comorbid disorders and social functioning in childhood. AB - OBJECTIVE: To examine the relationship of depressive, conduct, and comorbid disorders and social functioning in psychiatrically referred youths. METHOD: Subjects were 94 boys and 67 girls (mean age at initial assessment = 11.5 years) who were repeatedly evaluated with standardized instruments during a mean interval of 4.4 years. On the basis of their diagnoses during the follow-up, children were designated as having had depressive, conduct, or both (comorbid) disorders or other conditions. Two domains of social functioning were assessed: social competence and self-esteem. RESULTS: Longitudinal analyses revealed that at any given point in time, depressive, conduct, and comorbid disorders were associated with low social competence and depressive disorder also was associated with low self-esteem. At the approximate age of 15 years, on average, children with a history of conduct or comorbid disorders had lower social competence than did children with a history of depressive disorder, but these groups endorsed similar levels of self-esteem. CONCLUSION: Some areas of social dysfunction associated with comorbid depressive and conduct disorders appear to reflect mostly the effects of conduct disorder. The latter condition has a more severe and longer-term impact on children's social competence than does depression. In addition, whereas depression has an adverse effect on self-esteem, this effect appears to be temporary. PMID- 9204681 TI - Physicochemical basis of the universal genetic codes--quantitative analysis. AB - Quantitative mathematic models have been developed to correlate the fragment hydrophobicity contribution constants (faa) of 20 amino acids with the physicochemical properties (mu, Hb, and square root of MW) of the four bases (U, A, C, G) of the codons, or those of the anticodons. Using the general equation faa = a mu 1 + b mu 2 + c mu 3 + d square root of MW1 + e square root of MW2 + f square root of MW3 + g Hb1 + h Hb2 + i Hb3 + j, where 1, 2, 3 refer to the first, the second and the third base respectively, correlation coefficient of about 0.82 can be obtained for all 20 amino acids coded by 61 different triplet codes. These correlations are statistically highly significant, even though they do not take into account the involvement of various factors and peptidyl transferases. Furthermore, the reasons for the three stop codons are revealed. The graphic presentation of the codons and the amino acids coded separates the acidic and the basic, the aromatic and the heterocyclic amino acids into different quadrants of an octagon. This is in agreement with the ancient Chinese Ying-Yang theory embedded in the classical I-Ching. PMID- 9204682 TI - Enzymatic generation of complex peptides. PMID- 9204683 TI - The role of apoptosis in neurodegenerative diseases. PMID- 9204684 TI - Viral quasispecies and the problem of vaccine-escape and drug-resistant mutants. PMID- 9204685 TI - Immunotherapy for brain diseases and mental illnesses. PMID- 9204687 TI - Dopamine receptor diversity: molecular and pharmacological perspectives. PMID- 9204686 TI - Natural products and their derivatives as cancer chemopreventive agents. AB - This review summarizes currently available data on the chemopreventive efficacies, proposed mechanisms of action and relationships between activities and structures of natural products like vitamin D, calcium, dehydroepidandrosterone, coenzyme Q10, celery seed oil, parsley leaf oil, sulforaphane, isoflavonoids, lignans, protease inhibitors, tea polyphenols, curcumin, and polysaccharides from Acanthopanax genus. PMID- 9204688 TI - Novel and unusual nucleosides as drugs. PMID- 9204689 TI - Oxidative damage to DNA: formation, measurement, and biological significance. PMID- 9204690 TI - Metabolism of cyclic ADP-ribose: a new role for NAD+ glycohydrolases. PMID- 9204691 TI - The role of poly(ADP-ribosyl)ation. PMID- 9204692 TI - A discussion of mechanisms of NO genotoxicity: implication of inhibition of DNA repair proteins. PMID- 9204693 TI - Teenage sexual attitudes in China. AB - Not much is known about the sexual attitudes of Chinese teenagers. In this article we endeavor to address this void by examining the sexual attitudes of Chinese teenagers with survey data collected in Sichuan Province in 1988. Our analysis has two goals: first, to describe aggregate attitudes toward premarital sexual practices; and second, to identify the principal factors that influence these attitudes. To accomplish the second goal we estimate several regression equations with predictor variables known to influence teen sexual attitudes. Our analysis reveals major differences and similarities between China and the United States and indicates that China's teenagers are somewhat strongly opposed to teen sexual contact, but seem to be more understanding of others who so engage, despite strike laws and public morality forbidding it. PMID- 9204694 TI - Impact of future cigarette smoking scenarios on mortality of the adult population in the United States, 2000-2050. AB - The prevalence of cigarette smoking in the United States has declined over the past few decades. However, some leveling-off in prevalence rates has been observed in recent years, and the rate for teenagers and young adults has even turned upward. This paper considers four alternative scenarios of future cigarette smoking patterns in the United States for the population 25 and over and measures the impact these different scenarios would have on excess mortality due to smoking and on the sex and age distributions of deaths. Scenarios reflecting higher levels of smoking prevalence produce considerably more deaths than scenarios tied to lower levels. As many as two and one-half million excess deaths would take place in the decade of the 2020's if a high prevalence, rather than low prevalence, assumption proves correct. Even when a constant prevalence, assumption proves correct. Even when a constant prevalence assumption is compared with a moderately-declining prevalence assumption, as many as one million excess deaths would be generated during that decade alone. Lowering smoking prevalence rates would also change the population sex ratio by reducing deaths for males more than deaths for females, and by contributing to the aging of the population. The results are interpreted in terms of the overall impact of smoking on mortality and with regard to public and private policy decisions related to cigarette smoking. PMID- 9204695 TI - Psychosocial factors in blood pressure variation: a comparative study of young Samoans. AB - Traditional peoples contacting modern cultures frequently experience increased levels of blood pressure. The aim of this investigation was to identify some acculturation-related psychosocial factors which might contribute to those elevations. Young Samoans living in a traditional village, in modernizing American Samoa, and as migrants in Hawaii were studied. Casual blood pressure, anthropometric measurements, and extensive interview data were collected. The most important factor predicting variation in blood pressure was body mass (BMI). This finding was particularly evident among the acculturated and migrant Samoans. In those locations women's abandonment of breast feeding may contribute to their higher BMI. Among acculturated and migrant women, measures of social support favored lower blood pressures. Among men in all locations a greater number of close social ties was linked to higher blood pressures with the exception of ties with parents. Parental ties were linked to substantially lower blood pressures. A knowledge of Samoan traditions favored lower blood pressure among migrant men, while knowledge of American culture favored higher pressures. Coping strategies and complaint behavior were also significant contributors in all locations. PMID- 9204697 TI - The changing pattern of interracial marriage. AB - Recent studies have demonstrated an increased occurrence of interracial marriages in the United States, indicating important shifts in intergroup relations. The effectiveness of traditional theoretical approaches in explaining who marries whom, however, remains problematic. Recently, exchange explanations (which have typically assumed that the black partner in the union exchanges educational and economic accomplishments for the higher "status" of the white spouse) have been replaced by progressive theories emphasizing a trend away from ascriptive and toward achievement norms. We extend this approach by predicting an economic and educational gap between spouses in interracial marriages when compared with racially homogamous marriages. Using the 1980 and 1990 Public Use Microdata Sample, we find continuing evidence that racial barriers in mate selection are weakening. Further, people who intermarry, regardless of race or gender, tend to have higher educational and economic status than those in homogamous marriages. There is still limited support for the kinds of social exchanges between spouses that were implied in earlier sociological theories. We conclude that (1) socioeconomic differentials are not always consistent with the exchange perspective and (2) that recent trends are not systematically eroding these socioeconomic differentials in mate selection. PMID- 9204696 TI - Trends in the relationship between breastfeeding and postpartum employment in the United States. AB - It is widely assumed that employment and breastfeeding are relatively incompatible behaviors in the United States; yet recently both the incidence of breastfeeding and the incidence of postpartum employment increased. This paper examines the relationship between these trends from 1968-86 using data from the National Surveys of Family Growth. I find that these trends result from increases in the likelihood that a woman engages in both breastfeeding and postpartum employment. There has been an increase over time in the incidence and duration of women concurrently breastfeeding and working. However, the majority of employed women did not concurrently breastfeed, suggesting that conflicts between these behaviors still exist. PMID- 9204698 TI - Racial fertility differences: the role of female employment and education in wanted and unwanted childbearing. AB - This paper employs data from a merged sample of the National Surveys of Family Growth to examine how female employment status conditions the relationship between education and wanted and unwanted births among African American and white women. A rationale is presented for why a minority group status hypothesis that posits lower fertility among more highly educated African American women as compared to similar white women might find support in the case of wanted births and among certain women, including earlier birth cohorts. Our results provide some evidence for these ideas as well as evidence for a social characteristics hypothesis that predicts convergence of childbearing with rising education. However, persistently higher levels of unwanted births among African American women of all educational levels suggest that the dynamics of racial fertility differences are more complex than either of the hypotheses imply. PMID- 9204699 TI - Reproductive change in Sri Lanka: analysis of intermediate variables, 1982 and 1987. AB - This study examines the intermediate determinants of fertility in Sri Lanka by making use of the data collected in the 1982 Sri Lanka Contraceptive Prevalence Survey and the 1987 Sri Lanka Demographic and Health Survey. The analysis shows that the most important inhibitor of potential fertility is deliberate control. The marital structure of the population is also an important fertility-inhibitor, but lactational infecundability is increasingly becoming an unimportant contributor. The findings show the success of the family planning program in Sri Lanka, which propelled fertility to a substantial lower level. Achievement of the replacement level fertility by the turn of the century, set by the Sri Lankan government, would largely depend on the efforts to increase the quality and quantity of contraceptive use and the duration of breastfeeding. PMID- 9204700 TI - Sex ratio at birth deviations in modern Venezuela: the Trivers-Willard effect. AB - This study evaluates the impact of the Trivers-Willard (T-W) effect on human populations, using demographic data collected from vital registration data in Venezuela. The evaluation of the sex ratio at birth (SRB) and of fetal and infant deaths supports the existence of T-W effect in the Venezuelan population in extreme conditions. This T-W effect was observable in the SRB but not at later ages and is related to the marital status of the mother. The results indicate that the investment in females associated with environmental adversity is greater than the investment in males associated with good environmental conditions. PMID- 9204701 TI - The determinants of IUD discontinuation in China: a discrete-time competing risk model analysis. AB - This research examines the social, demographic, and family-planning-program factors that influence the occurrence of IUD discontinuation among Chinese women, using a sample of 14,639 IUD use segments from the 1988 Chinese National Survey of Fertility and Contraceptive Prevalence. A discrete-time competing-risk event history method is employed to identify the determinants of IUD discontinuation by five kinds of reasons: contraceptive failure, expulsion, switching method, side effects and other nonmethod-related reasons. The predictors of IUD discontinuation suggest that a number of mechanisms are in operation. Some of the determinants may reflect the effects of the family planning program; some may illustrate women's physiological and biological reactions to IUD's; some may be related to women's previous history of contraceptive use; and still others may indicate social characteristics of women that lead them to have their IUD's removed. PMID- 9204702 TI - Common themes in translational and transcriptional regulation. AB - How transcription and translation initiation complexes are assembled and regulated has been the subject of research for many years. New information on the translation initiation complex has revealed similarities between its organization and regulation with that of the transcription initiation complex. Here, we will summarize these similarities in order to set up a framework for future integration of results from each of these areas. PMID- 9204703 TI - SPRY domains in ryanodine receptors (Ca(2+)-release channels). PMID- 9204704 TI - A repetitive sequence in subunits of the 26S proteasome and 20S cyclosome (anaphase-promoting complex). PMID- 9204705 TI - Mitotic repression of the transcriptional machinery. AB - Nuclear RNA transcription is silenced when eukaryotic cells enter mitosis. Until recently, this repression was thought to derive solely from the condensation of interphase chromatin into mitotic chromosomes. Recent studies, however, have shown that changes in chromatin structure and occupancy of promoter elements by both general and gene-specific transcription factors also play a role in transcriptional silencing. In addition, studies with simplified systems reveal that reversible phosphorylation of the basal transcriptional machinery represses transcription at mitosis. PMID- 9204706 TI - Transporters of nucleotide sugars, nucleotide sulfate and ATP in the Golgi apparatus. AB - Proteins and glycolipids are glycosylated, sulfated and phosphorylated in the lumen of the Golgi apparatus. The nucleotide substrates of these reactions must first be translocated from the cytosol into the Golgi lumen by specific transporters (antiporters). These are hydrophobic, transmembrane spanning proteins that appear to regulate post-translational modifications in the Golgi lumen. PMID- 9204707 TI - Redox signal transduction via iron-sulfur clusters in the SoxR transcription activator. AB - Protein iron-sulfur (FeS) centers have recently been implicated in the regulation of gene expression. In the redox-sensing SoxR protein, the oxidation state of [2Fe-2S] centers controls its activity as a transcription activator independent of DNA-binding ability. Thus, FeS centers allosterically link cellular oxidative stress to the expression of defense genes. PMID- 9204708 TI - Structural and functional considerations of the aminoacylation reaction. AB - Aminoacyl-tRNA synthetases (aaRS) bind their substrates-ATP, amino acids and tRNA and stabilize putative transition states in the aminoacylation reaction. Here, we discuss the common and distinguishing structural and functional themes of the 20 known aaRS, which can be divided into two main classes (I and II) and into further subgroups on this basis. PMID- 9204709 TI - Tailoring cAMP-signalling responses through isoform multiplicity. AB - Multiple forms of cAMP phosphodiesterases (PDE), adenylate cyclase and protein kinase A (PKA) allow cells to tailor the responsiveness of the cAMP-signalling system and to allow for its dynamic adjustment. Multiple forms of these enzymes confer spatial and temporal characteristics on cAMP signalling so as to affect compartmentalised responses within a single cell type. The ability to breach the PKA activation threshold can depend upon either or both the activation of adenylate cyclase and inhibition of specific PDE isoforms. PMID- 9204711 TI - Internet resources on G-protein-coupled receptors. PMID- 9204710 TI - Optimizing multiplex and LA-PCR with betaine. AB - Methods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.molbio.methds-reagnts, available on the internet. This month's column discusses the use of additives for optimizing the amount and quality of product obtained through multiplex and 'long and accurate' polymerase chain reaction (LA-PCR). For details on how to partake in the newsgroup, see the accompanying box. PMID- 9204713 TI - Tropical medicine and traditional methodologies: maximizing options for safe and effective health care coverage. PMID- 9204712 TI - Enzymes and symmetrical molecules. PMID- 9204714 TI - The scope and the limits of traditional care. PMID- 9204715 TI - Herbal remedies for malaria. PMID- 9204716 TI - Plants as a source of antimalarial drugs. PMID- 9204717 TI - Antimalarial plants used by Hausa in northern Nigeria. PMID- 9204718 TI - Herbal treatment of malaria--four case reports from the Rukararwe Partnership Workshop for Rural Development (Uganda). AB - We present a small study of four cases of malaria treated using a traditional herbal remedy at Rukararwe, Uganda. Our results demonstrate that this remedy has the potential to cure malaria clinically and parasitologically. PMID- 9204719 TI - Malaria and antimalarial plants in Roraima, Brazil. AB - One of the numerous problems created by the gold rush which took place in northern Brazil (Roraima State) at the end of the 1980s was a severe epidemic of malaria amongst the indigenous peoples of the region. Worst hit were the Yanomami Indians, who had lived in almost total isolation prior to this event. The problem has been exacerbated by the development of chloroquine-resistant strains of Plasmodium falciparum. In an effort to identify viable alternatives to dependence on western medicine for malaria treatment, a survey was carried out on the local plant species (wild and cultivated) used for this purpose in Roraima. Fieldwork was carried out amongst seven indigenous peoples, as well as with the non indigenous settlers. Over 90 species were collected, many of which have been cited as used for treatment of malaria and fevers elsewhere. Knowledge of antimalarial plants was found to vary greatly between the communities, and in some cases there was evidence of recent experimentation. Initial screening of plant extracts has shown a high incidence of significant antimalarial activity amongst the species collected. PMID- 9204721 TI - Phytolacca dodecandra: a multipurpose plant for control of vector-borne diseases. PMID- 9204720 TI - Control of water-borne parasitic diseases with natural products: the potential of Dialium guineense as a molluscicide. AB - The molluscicidal activity of the fruit and leaves of Dialium guineense was found to be due to glycosides of the triterpenoid oleanolic acid. Three glycosides were isolated from the fruit and a fourth from the leaves and are known compounds. The amount of total saponins present in D. guineense makes it a good candidate for a readily available molluscicide in Nigerian villages. PMID- 9204722 TI - Global curriculum for wound management. PMID- 9204723 TI - The effects of honey on Leishmania parasites: an in vitro study. AB - The activities of honey dilutions were investigated against three species of Leishmania. The results were compared with the effects of the same concentrations of sugar. Honey and sugar both have anti-leishmanial effects in vitro, but our results indicated that honey is superior to sugar. PMID- 9204724 TI - Traditional medicine and the infrastructure for burn care in Vietnam. PMID- 9204725 TI - Traditional herbal medicines used to treat wounds and injuries in Nepal. AB - In rural Nepalese societies, due to the lack of modern health services and facilities, folk herbal preparations are still the dominant method of therapy for common ailments. These remedies are fairly well accepted, easily available, bear a minimal cost and are generally the only available resource. Information on the curative properties of 42 plant species from 40 genera and 23 families that are used to treat wounds and injuries, has been documented. These herbal remedies are based on ancestral knowledge and personal experience. Their successful use indicates that they are alive and functioning in the rural localities. Although the techniques of preparation of drugs or dosage forms employed by traditional healers have generally been observed to be poor and in most cases do not comply with the requirements of modern pharmaceutical practices, there are trends towards the development and modernization of practice including the standardization of specific dosage, storage of drugs, use of specific types of containers and their labelling. A critical evaluation of the phytochemistry and pharmacology of the alleged curative plants and the traditional pharmaceutical practices employed has been strongly recommended. In rural Nepalese societies, the number and distribution of traditional herbal practitioners and faith healers greatly exceed all other health workers. Their influence should be applied to the task of motivating the rural populace to adopt authentic herbal remedies and other health-related hygienic behaviour. PMID- 9204726 TI - Studies of the microcirculation in China. PMID- 9204727 TI - Traditional health practitioners as primary health care workers. AB - The author conducted a field study in 1993 to evaluate the effectiveness of four projects that were training traditional health practitioners (THPs) to provide primary health care (PHC) services in Ghana, Mexico, and Bangladesh. The study, funded by a grant from the World Health Organization, Division of Strengthening Health Services, concluded that incorporating trained THPs in PHC programmes can be cost effective in providing essential and culturally relevant health services to communities. The main objective of the study was to evaluate how effective the training projects were and to determine what impacts they might have upon the communities served. A qualitative field evaluation was performed using data collected from project documents, observations, and field interviews with a selection of health agency staff, THPs, and community members. A summary of results is presented from the four field studies. For details refer to the full report. PMID- 9204728 TI - The participation of African traditional healers in AIDS/STD prevention programmes. PMID- 9204729 TI - Umbanda healers as effective AIDS educators: case-control study in Brazilian urban slums (favelas). AB - During a 12-month period (November 1994-October 1995), Afro-Brazilian Umbanda healers (Pais-de-Santo) taught 126 fellow healers from 51 Umbanda centres (terreiros) located in seven overcrowded slums (favelas) (population 104-343) in Brazil's northeast, the biomedical prevention of AIDS, including safe sex practices, avoidance of ritual blood behaviours and sterilization of cutting instruments. A face-to-face educational intervention by healers, marginalized in society yet respected by devotees, which blended traditional healing-its language, codes, symbols and images- and scientific medicine and addressed social injustices and discrimination was utilized in this project supported by the Brazilian Ministry of Health, National Program in STDs/AIDS. Significant increases (P < 0.001) in AIDS awareness, knowledge about risky HIV behaviour, information about correct condom use, and acceptance of lower-risk, alternative ritual blood practices and decreases (P < 0.001) in prejudicial attitudes related to HIV transmission were found among mobilized healers as compared to 100 untrained controls. Respected Afro-Brazilian Pais-de-Santo can be creative and effective partners in national HIV prevention programmes when they are equipped with biomedical information about AIDS. PMID- 9204730 TI - Surgery of tuberculosis: state of the art--introduction. PMID- 9204731 TI - Surgery for pulmonary tuberculosis. AB - During the period 1990-1994 a total of 578 operations were performed in 502 patients with various forms of tuberculosis. Most of the patients (68%) were men aged 20 to 50 years (70%). Sputum cultures were positive in 55% of the patients. More than half of all patients were chronic smokers, and about 10% were alcoholics or drug addicts. There were no human immunodeficiency virus-infected patients, and none with acquired immunodeficiency syndrome. The most frequent surgical interventions were, according to the classification adopted in Russia, for cavernous or fibrocavernous tuberculosis (196 cases) and tuberculomas (161 cases). The main operative procedures used were pulmonary resection (n = 280) and pneumonectomy or pleuropneumonectomy (n = 80). Diseased intrathoracic lymph nodes were ablated in 62 patients. Thoracoplasty or thoracomyoplasty were performed in 46 cases, thoracostomy in 37, closure of a thoracic wall defect in 27, and reamputation of the main bronchial stump in 6. Postoperative complications arose in 20% of the patients. More than half occurred in the pleural cavity or bronchi and were associated with tuberculous infection. The postoperative hospital case fatality rate was 2%. The overall clinical efficacy by the time of discharge was 82.7% (95% in tuberculomas). Reactivation of tuberculosis over the first 3 years after discharge occurred in 6.6% of the patients. Most patients with large or multiple caverns, tuberculomas, intrathoracic caseous lymphadenitis, or various complications of pulmonary tuberculosis cannot be cured (or are not amenable to care in principle) by means of antibacterial therapy because of irreversible morphologic changes in the lungs, bronchi, pleura, lymph nodes, or thoracic wall. For this reason, indications for surgical management of pulmonary tuberculosis should be generally expanded. Excessively long antibacterial therapy for tuberculosis is often inadvisable. Although the availability of standardized regimens of antibacterial therapy is strategically essential, each patient must be treated according to an individual plan. In certain cases thoracic surgeons should be enlisted to participate in the development of such plans. PMID- 9204733 TI - Surgical indications for treatment of pulmonary tuberculosis. AB - Surgery for pulmonary tuberculosis (PTB) has passed through various stages throughout history, having been the treatment of choice in the past. It has now been relegated to second place for treatment of this disease. One of the most strongly debated surgical indications has been clinical picture of multidrug resistance with the focus of pulmonary tuberculous activity located in a segment, lobe, or lung. In these cases some authors have described good results with surgical excision. Another important indication is the complications of PTB, among which bronchiectases (provoking pictures of suppuration, superinfections, or hemoptysis) are found, along with known destructive pulmonary sequelae such as destroyed lung, massive hemoptysis, and the presence of a bronchopleural fistula that cannot be resolved with pleural drainage. The presence of a neoplasm in a patient affected by PTB is a surgical indication if the lesion is resectable. The existence of an unidentifiable pulmonary mass or node is a surgical criterion because it might signal bronchogenic carcinoma. A frequent indication for surgery is pulmonary aspergilloma, which in a large percentage of cases is a destructive PTB sequela and generates serious complications, hemoptysis being the most frequent. Mediastinal tuberculous lymphadenitis that produces compressive symptoms and pulmonary complications, especially in children, is another surgical indication for decompressing the bronchial tree. The surgery in these cases consists in excision and curettage of adenopathies. Surgery therefore now constitutes a valid option for the treatment of certain clinical pictures of PTB that do not respond to medical treatment, are serious, and are potentially fatal. PMID- 9204732 TI - Role of thoracic surgery for childhood tuberculosis. AB - Lymphadenopathy is the hallmark of intrathoracic tuberculosis in children. The role of the thoracic surgeon in treating childhood tuberculosis is to relieve the more severe symptoms of lymphadenopathy, prevent the more long-term secondary damage that lymphadenopathy may cause to the lung, and treat the sequelae of thoracic tuberculosis. We reviewed the role of surgery in childhood tuberculosis at Red Cross Children Hospital from January 1981 to January 1996 in 161 children under 13 who were admitted for 168 therapeutic surgical interventions for proved intrathoracic tuberculosis and its related complications. We classified patients according to the pathophysiology of their disease to clarify the role of surgery in their management. Successful decompression of lymph nodes that were acutely compromising major airways was done in 25 children, and decompression for chronic airway compression was successful in 8 of 11 children. Therapeutic bronchoscopy successfully opened an airway obstructed by intraluminal tissue in 68% of 28 patients, with long-term pulmonary reexpansion in 50%. Pulmonary resections for postprimary tuberculous damage were done in 72 patients with a mortality of 2.7% and morbidity of 16.7%. Another 17 patients were operated on for pleural disease and 15 for other tuberculosis-related problems. The mortality for all patients undergoing surgery for complications of tuberculosis during childhood was 1.9% (3/161), suggesting that when indicated, an aggressive surgical approach is relatively safe. PMID- 9204734 TI - Treatment of bronchial stricture due to endobronchial tuberculosis. AB - Between 1974 and 1995 we encountered 19 cases of bronchial stricture or obliteration caused by endobronchial tuberculous lesions. In 11 the involvements were located at the right bronchus (including involvements of segmental and middle lobe bronchi) and in 8 at the left bronchus. On bronchoscopic biopsy of the stenosed bronchus, 7 patients showed histopathologic findings of tuberculous bronchitis, but 12 patients showed nonspecific inflammatory granular tissue. Five patients were kept under conservative observation because of mild subjective symptoms or refusal to undergo operation. Two patients underwent stent procedures but had poor outcomes. Twelve patients underwent operation. As the bronchial lesions in four of them were confined to the lobar or segmental bronchus, lobectomy was performed. One patient with a history of infantile tuberculosis had developed complete obliteration of the left main bronchus and cystic bronchiectasis in the entire lung parenchyma; pneumonectomy was essential. Seven patients who had strictures involving the main bronchus underwent bronchoplastic surgery with right (n = 4) or left (n = 3) upper sleeve lobectomy. None of the patients treated surgically showed any postoperative complication or recurrence of the tuberculosis. These surgical results for endobronchial tuberculosis indicate the need for early detection and operation. Bronchoscopy and computed tomography are the methods of choice for accurate diagnosis of bronchial involvement and assessment of the surgical indications. It is emphasized that bronchoplastic surgery is the best treatment for bronchial stricture involving bilateral main bronchi. PMID- 9204735 TI - Modern morbidity following pulmonary resection for postprimary tuberculosis. AB - Between January 1991 and March 1996, a total of 28 patients with postprimary tuberculosis underwent resection for disease progression (n = 8), multidrug resistance or noncompliance to the medical treatment (n = 11), parenchymal sequelae (n = 3), suspected cancer (n = 5), and for the correction of postpneumonectomy bronchopleural fistula and empyema (n = 1). On admission, eight patients presented with sputum positivity (28.6%). Similar to previous series, tubercular predilection for upper lobes was confirmed (21/28, 75%); accordingly, upper lobectomy through an extrapleural approach was the most common procedure (16/28, 57.1%). Atypical segmental resections or segmentectomies were performed in seven patients (25%), whereas a bilobectomy was necessary in another three patients (10.7%) and a completion pneumonectomy in one (3.6%). Additional procedures were an open-window thoracostomy with transpericardial closure of the main bronchus and a tailored thoracoplasty. No operative mortality was reported. Healing was achieved in 26 patients (93%). Bleeding, either from the chest wall or hilar dissection, was the only reported intraoperative complication. Median blood loss, inclusive of early postoperative collections from chest tubes, reached 1330 ml (range 100-3700 ml). Major postoperative complications included recurrent disease (2/28, 7%) in sputum-positive patients and segmental pulmonary embolism (3.5%). Causes of minor morbidity were air leaks resulting in residual space undergoing spontaneous resolution (18%), wound breakdown (14%), and, fever (11%). This limited series confirms the therapeutic value of the surgical treatment of postprimary tuberculosis, provided that correct indications, adequate pre- and post-operative medical coverage, and meticulous technique are applied. PMID- 9204736 TI - Abdominal tuberculosis. AB - Throughout the world tuberculosis is associated with poverty, deprivation, and human immunodeficiency virus infection. Abdominal tuberculosis is usually of insidious onset with diverse symptoms and signs. A few present with acute complications of perforation, obstruction, or bleeding. The diagnosis is difficult, especially in areas where the disease is less common, as many patients do not have evidence of pulmonary tuberculosis or a positive skin test. The main differential diagnosis ranges from Crohn's disease in the young and advanced malignancy in the elderly. Delayed diagnosis is common, resulting in high mortality. Many investigations provide findings suggestive but not diagnostic of tuberculosis. With peritoneal tuberculosis, assay of ascitic fluid adenosine deaminase activity is a valuable, simple method of diagnosis that may reduce the need for laparoscopic biopsy. If the clinical suspicion of abdominal tuberculosis is high, a trial of medical treatment is appropriate. Surgery should be reserved for the complications of the disease. All patients require treatment with three antituberculous drugs over a 6-month course. PMID- 9204737 TI - Large bowel tuberculosis and possible influencing factors for surgical prognosis: 30 years' experience. AB - The incidence of tuberculosis is rising in the United States. Similarly, the incidence of pulmonary tuberculosis in Taiwan is increasing, but that of large bowel tuberculosis in this region has not been reported. The purpose of this study was to investigate the changing disease pattern and to determine some possible surgical prognostic factors for large bowel tuberculosis. Seventy cases of large bowel tuberculosis treated at our institute during the period 1965-1995 were reviewed and analyzed. A steady decline in the case number of large bowel tuberculosis were noted from 1975, but there seems to be a slight increase in cases since 1990. The average age of these patients was 65.1 years, and none had human immunodeficiency virus infection. The ileocecum is the most common region of involvement. Of these 70 patients, 59 had not been definitively diagnosed until surgery. Active pulmonary tuberculosis was found in 18 patients (25.7%). The incidence of postoperative pulmonary complications was higher in patients with active pulmonary tuberculosis or disseminating large bowel tuberculosis. Postoperative abdominal complications, including intestinal obstruction, abdominal cutaneous fistula, and wound infection, were seen in 13 patients, none of whom had active pulmonary tuberculosis. Although the incidence of tuberculosis has been reduced for years, it is now rising. Physicians should bear in mind the possibility of large bowel tuberculosis in patients with intestinal obstruction without specific origin. Postoperative respiratory care is important for patients with pulmonary tuberculosis, with either active or disseminating lesions. PMID- 9204738 TI - Indications for surgical management of genitourinary tuberculosis. AB - The incidence of tuberculosis has risen in many parts of the world, and more attention is being focused on genitourinary tuberculosis (GT), the second most common extrathoracic form of tuberculosis. Although chemotherapy is the mainstay of treatment, ablative surgery as a first-line management may be unavoidable for sepsis or abscesses. In cases with hydronephrosis and progressive renal insufficiency caused by obstruction, renal drainage (by stenting or nephrostomy) must be performed immediately. In all other situations triple-drug chemotherapy should be undertaken for at least 6 months and stable conversion obtained before ablative or reconstructive surgery is planned. Nephrectomy or partial nephrectomy is indicated for nonfunctioning or poorly functioning kidneys, particularly if continuous flank pain or hypertension is present. Stenosis of the ureter usually can be managed by temporary stenting and adjuvant corticosteroid therapy. Today the indications for augmentation are rare, but bladder replacement may be combined with ureter replacement using segments of intestine. PMID- 9204739 TI - Reconstructive surgery for treatment of urogenital tuberculosis: 30 years of observation. AB - Tuberculosis has continued to be a public health problem around the world. The urogenital tuberculosis clinic in the Russian Scientific Research Institute of Phthisiopulmonology was founded in 1950. The development of reconstruction operations for those with urogenital tuberculosis began in 1960. Since then 4298 patients with urogenital tuberculosis have been treated, and 2364 operations have been performed: 885 to remove an organ, 531 to preserve an organ, and 948 for reconstruction. The cases of extrapulmonary tuberculosis in recent years have increased to 6.0%. Surgery for urogenital tuberculosis is performed after specific medical therapy has been tried, but it is difficult, particularly if it is a reconstruction. The clinical features and results of various ureteral neoimplantation procedures using intestinal transplants (ileocystoplasty, sigmoidocystoplasty, cecocystoplasty) are discussed. PMID- 9204740 TI - Management of mycobacterial cervical lymphadenitis. AB - The treatment results of mycobacterial cervical lymphadenitis in 69 patients between 1990 and 1993 are reviewed. All patients underwent surgical procedures consisting of total excision or selective nodal dissection for lymphadenopathies and curettage for fluctuant cases, followed by antituberculous chemotherapy applied according to the likely or proved mycobacterial species. For this purpose, three or four drugs (including isoniazid, rifampin, ethambutol, and streptomycin) were used for 12 to 18 months. The cure rate was 100% after a minimum follow-up of 3 years. Clinical features, treatment modes, and guidelines for management are discussed. PMID- 9204741 TI - Surgical treatment of bacillus Calmette Guerin lymphadenitis. AB - Although its protective effect is contested and the risk of contracting tuberculosis is rather low nowadays, BCG vaccination is frequently performed. Changes of strain repeatedly led to an increased complication rate. In Austria between 1990 and 1991, of 3386 newborn babies (Strain Pasteur) 116 developed lymphadenitis 3 to 28 weeks after vaccination. The affected children received four types of treatment: nothing specific, isoniazid, or surgery with and without isoniazid. Surgical treatment was found to be necessary in 96 cases. Bacilli were successfully grown in culture in 46% of cases up to week 20 after vaccination; but later than 20 weeks no culture became positive. All cultured bacteria were isoniazid-sensitive. From our data we drew the following conclusions: isoniazid therapy did not prove successful when inflamed lymph nodes exceeded a certain size. Suppurative lymphadenitis in lymph nodes exceeding 1.0 to 1.5 cm usually led to infiltration or even perforation of the skin. Surgery prevents these complications and significantly reduces healing time. Adjuvant isoniazid therapy cannot be recommended, except for generalized BCG tuberculosis. PMID- 9204742 TI - Prevention of postoperative late kyphosis in Pott's disease by anterior decompression and intervertebral grafting. AB - A total of 185 patients with Pott's disease were operated on between 1973 and 1992. Anterior decompression by preserving the pleura (extrapleural approach) was the preferred method in the thoracic spine. In the lumbar spine the approach was retroperitoneal, and interbody fusion was performed in both for the thoracic and the lumbar regions. Anterior decompression and intervertebral grafting comprised the treatment. In five patients, internal fixation accompanied anterior decompression and intervertebral grafting. The aim of instrumentation was to enhance anterior spinal stability, and Alici spinal instrumentation was the preferred device. Graft destruction and a late increase in kyphosis was prevented by this means. The mean follow-up period was 7.5 years. Thirty-two of the cases were admitted to the clinic because of Pott's paraplegia: 19 of the cases recovered completely following anterior decompression; partial recovery was observed in 5 cases; but 3 cases did not recover. Various complications, including seven deaths, were observed in 42 of the cases. PMID- 9204743 TI - Risks of the minimal access approach for laparoscopic surgery: multivariate analysis of morbidity related to umbilical trocar insertion. AB - The objective of this study was to determine the morbidity associated with trocar and needle insertion for laparoscopic surgery and to identify risk factors for complications. Data from a prospectively collected database of all laparoscopic operations performed at a major teaching hospital over a 4-year period were analyzed. In 203 patients closed laparoscopy (Veress needle plus blind trocar insertion) was used to establish the pneumoperitoneum. Open laparoscopy with a Hasson's trocar was performed in 200 patients. A total of 1206 operative trocars were inserted (mean +/- SD 2.99 +/- 0.4). Sixty-nine percutaneous punctures for cholangiography or liver biopsy were carried out. Of the 403 patients undergoing laparoscopic surgery, 20 (3%) had developed complications specifically related to the access to the abdominal cavity after a minimum follow-up of 3 months, abdominal wall hematoma being the most frequent (n = 8, 2.0%), followed by umbilical hernias (n = 6, 1.5%) and umbilical wound infection (n = 5; 1.2%). The rate of penetrating injuries was 0.2% (n = 1). Of 20 complications, 15 (75%) were related to the umbilical insertion site. Female sex and closed laparoscopy were associated with umbilical morbidity by univariate analysis. In a multivariate analysis, closed laparoscopy was the only factor associated with these complications (odds ratio = 6.0; p = 0.04). Age, gender, obesity, diabetes mellitus, previous abdominal surgery, and the specific procedure had no influence. In conclusion, gaining access to the peritoneal cavity for laparoscopic surgery may cause severe complications, most of which are related to the umbilical trocar. Although closed laparoscopy can be safely used, open laparoscopy is associated with a lower morbidity rate; therefore its utilization is recommended. PMID- 9204744 TI - Emergency minilaparotomy cholecystectomy for acute cholecystitis: prospective randomized trial--implications for the laparoscopic era. AB - This prospective controlled trial evaluates the efficacy of minicholecystectomy (MC) in cases of acute cholecystitis compared to that of conventional cholecystectomy (CC) and discusses its implications in the laparoscopic era. Sixty consecutive patients with acute cholecystitis were prospectively randomized into two groups: MC group (30 cases) and CC group (30 cases). The two groups were well matched with regard to age, sex, weight/height index, previous upper abdominal surgery, and APACHE II scores. The mean length of incision was 5.5 cm (range 4.5-9.0 cm) in the MC group compared to 13.5 cm (range 12-16 cm) in the CC group. No significant differences were found between MC and CC with regard to operative time (69.1 +/- 17.0 and 68.1 +/- 15.4 minutes, respectively; p = 0.82), operative difficulty on a 1 to 10 scale (5.2 +/- 1.5 versus 4.6 +/- 1.6, respectively; p = 0.177), and complication rate (11% and 17%, respectively; p = 0.19). Significantly lower analgesia requirements were noted in the MC group: 27.5 +/- 14.6 mg of morphine sulfate compared to 44.5 +/- 13.2 mg in the CC group (p < 0.001). In addition, the duration of hospital stay was significantly shorter for MC patients (3.1 +/- 1.0 days) than in CC patients (4.7 +/- 1.2 days) (p < 0.001). Twenty-two patients (73.3%) in the MC group were reported to return to normal daily activities 2 weeks after the operation, compared to only 12 (40%) in the CC group (p = 0.0028). MC is safe and applicable as an emergency procedure for acute cholecystitis. It is superior to CC in terms of convalescence and cosmesis. The results of MC in the setting of acute cholecystitis compare favorably with the published results of laparoscopic cholecystectomy. PMID- 9204745 TI - Laparoscopic cholecystectomy for acute cholecystitis: prospective trial. AB - This prospective study determines the indications for and the optimal timing of laparoscopic cholecystectomy (LC) following the onset of acute cholecystitis. It also evaluates preoperative and operative factors associated with conversion from laparoscopic cholecystectomy to open cholecystectomy in the presence of acute cholecystitis. Having been established as the procedure of choice for elective cholelithiasis, LC is now also used for management of acute cholecystitis. Under these circumstances the procedure may be difficult and challenging. Certain favorable and unfavorable conditions may be present that influence the conversion and complication rates. Information about these conditions may be helpful for elucidating the optimal circumstances for LC or when the procedure is best avoided. We performed LC on an emergency basis as soon as the diagnosis was made on all patients presenting with acute cholecystitis from January 1994 to December 1995. All preoperative, operative, and postoperative data were collected on standardized forms. Of the 137 patients registered, 130 were eligible for the audit. Seven patients found by laparoscopic intraoperative cholangiography to have choledocholithiasis were converted for common bile duct exploration and were excluded from the study. Altogether 83 patients (72%) underwent successful LC and 37 (28%) needed conversion to open cholecystectomy. The conversion rate of acute gangrenous cholecystitis (49%) was significantly higher than that for uncomplicated acute cholecystitis (4.5%) (p < 0.00001) and for hydrops (28.5%) and empyema of the gallbladder (28.5%) (p = 0.004). The difference in conversion between the group with acute necrotizing (gangrenous) cholecystitis and the two groups with hydrops and empyema of the gallbladder was not statistically significant (p = 0.07). The complication rates of acute cholecystitis, hydrops, empyema of the gallbladder, and gangrenous cholecystitis were 9.0%, 9.5%, 14.0%, and 20.0%, respectively (p = NS). Patients with an operative delay of 96 hours or less from the onset of acute cholecystitis had a conversion rate of 23%, whereas a delay of more than 96 hours was associated with a conversion rate of 47% (p = 0.022). The complication rate was 8.5% in the laparoscopic group and 27% in the converted group (p = 0.013). Patients over 65 years of age, with a history of biliary disease, a nonpalpable gallbladder, WBC count over 13,000/cc, and acute gangrenous cholecystitis were independently associated with a high LC conversion rate; male patients, finding large bile stones, serum bilirubin over 0.8 mg/dl, and WBC count over 13,000/cc were independently associated with a high complication rate following laparoscopic surgery with or without conversion. Generally, LC can be performed safely for acute cholecystitis, with acceptably low conversion and complication rates. Different forms of cholecystitis carry various conversion and complication rates in selected cases. LC for acute cholecystitis should be performed within 96 hours of the onset of disease. Predictors of conversion and complications may be helpful when planning the laparoscopic approach to acute cholecystitis. PMID- 9204746 TI - Selective treatment of differentiated thyroid carcinoma. AB - Over a period of 20 years 84 papillary and 82 follicular carcinomas operated on by one surgeon and examined by one pathologist were documented prospectively, treated selectively, and followed for 1 to 20 years (median 7 years). Tumors with a low risk of recurrence or incurable disease-i.e., papillary carcinoma pT1-3 N0 M0 (n = 56) and minimally invasive follicular carcinoma (n = 37)-were treated by a limited-radicality hemithyroidectomy or total thyroidectomy without radioiodine in 79 of the 93 cases (85%). No unfavorable course was observed, and only one curable recurrence (1.3%) developed contralaterally after hemithyroidectomy for papillary cancer. Of the remaining 73 patients, including 100% of those with nodal involvement, 65 (89%) underwent total thyroidectomy with radioiodine. Total thyroidectomy was achieved in 34% of the cases by completion thyroidectomy, based on definitive histologic examination. No instance of a serious, potentially incurable recurrence and no tumor-related death was observed in patients with a papillary TNM stage I+II or with a minimally invasive follicular carcinoma. Five of the patients (6%) with papillary carcinoma, all with TNM stage III or IV, and seven of the patients (8.5%) with follicular carcinoma, all grossly invasive and pT3 or pT4, had tumor-related deaths following total thyroidectomy in all and with remnant ablation in 10 cases. A potentially curable node recurrence occurred in two patients 1 and 10 years, respectively, after primary treatment. Permanent hypoparathyroidism (n = 4) (2.4%) and permanent recurrent laryngeal nerve palsy (n = 2) (1.2%) were observed only in patients with a grossly invasive follicular carcinoma and concomitant benign recurrent goiter. We conclude that (1) hemithyroidectomy or total thyroidectomy without radioiodine is adequate for papillary carcinoma pT1-3 N0 and minimally invasive follicular carcinoma; (2) there were no nodal recurrences in tumors recognized as node-negative; and (3) extracapsular excision of one or both lobes can be carried out technically with low morbidity. The study confirms the prognostic value of age-related TNM classification for papillary carcinoma; classification of follicular thyroid carcinoma as minimally invasive or grossly invasive proved to be useful. PMID- 9204747 TI - Parathyroid function and histology in patients with parathyroid adenoma: correlation of clinical and morphologic findings. AB - Serum levels of parathyroid hormone (PTH), alkaline phosphatase (ALP), calcium, creatinine, and vitamin D and the glomerular filtration rate were compared with the histologic properties and expression of PTH and chromogranin A in excised parathyroid adenomas from patients with primary hyperparathyroidism (pHPT). PTH and chromogranin A were detected immunohistochemically, and their mRNA was demonstrated by in situ hybridization with quantification of their mRNA levels by image analysis. There was a positive correlation between the cellular levels of PTH mRNA and the cellular levels of chromogranin A mRNA (r = 4.4; p < 0.05). However, within certain parts of the adenomas, mostly consisting of chief cells, the expression of PTH mRNA and chromogranin A mRNA was heterogeneous and the levels did not correspond to each other. A reduced suppressibility of PTH in patients with pHPT was confirmed. Although cellular levels of PTH and chromogranin A and their mRNAs were low in the oxyphilic parts of the adenomas, there was no correlation between the amount of oxyphilic cells in the adenomas and the suppressibility of PTH by calcium. There was also no association between the cellular levels of PTH mRNA or chromogranin A mRNA as studied by image analysis and "calcium sensitivity." Our results thus demonstrate that although PTH and chromogranin A mRNA levels are in general correlated to each other there are differences in their expression within and between individual parathyroid adenomas. It therefore seems likely that the expression of PTH and chromogranin A are differentially regulated, and that PTH and chromogranin A may not always be co-secreted. This point could be of importance, as chromogranin A and its cleavage products are known to influence PTH secretion. PMID- 9204748 TI - Function of "nontrauma" surgeons in level I trauma centers in the United States. AB - Although the general "trauma" surgeon is usually the team leader in level I trauma centers, the use of surgical subspecialists and nonsurgeons is frequently ill-defined. This study was done to gain data in regard to actual use of subspecialists in busy centers. First, a survey of the patterns of staffing in 140 trauma centers was elicited by mail questionnaire, supplemented by telephone cells. Second, records of 400 consecutive patients at the Elvis Presley Trauma Center were reviewed to determine the use of subspecialists during the first 24 hours of care of individual patients. There were differences in the use of surgical subspecialists and nonsurgeons at different centers: in receiving, admitting, operating, and critical care areas and in privileges for admission and attending of inpatients. Consultation "guidelines" are used for many specific injuries. At our center, a mean of 1.92 subspecialists, in addition to general surgeons, were involved in the early care of each patient. Problems exist in many centers regarding the use of subspecialists, especially for management of facial and chest injuries. In some centers nonsurgeons function in the intensive care unit, and as admitting and attending physicians of trauma patients. PMID- 9204749 TI - Little-known aspect of Theodor Billroth's work: his contribution to musical theory. AB - Theodor Billroth's contribution to musical theory is discussed and evaluated. Billroth was a close friend of the composer Johannes Brahms and was himself extremely musical. At his death he left the manuscript of a book on musical theory, Wer ist-Musikalisch?, which had gone through four editions by 1912. In it he attempted to answer important questions on the nature of sound perception, the importance of rhythm as a fundamental element in music, the relation of pitch, tone, and volume, and the ways in which to account for the affective power of music. This article outlines the main concepts, contributions, and opinions offered by Billroth. PMID- 9204750 TI - Forest tree biotechnology. AB - The forest products industry has traditionally viewed trees as merely a raw, and more or less immutable, natural resource. However, unlike such inanimate resources as metallic ores, trees have the potential to be modified genetically, essentially transmuting lead into gold. Increasingly, modern alchemists are applying the tools of biotechnology in efforts to reduce the biological constraints on forest productivity. Several new methodologies being used to address problems in forest biology are described with respect to their potential impact on forest tree improvement. In addition to addressing problems inherent to the current use of trees for production of pulp and paper or solid wood products, genetic manipulation of trees brings with it the potential to create new industries based on the novel characteristics of transgenic trees, e.g. trees containing transgenes to detoxify specific pollutants could be used in the remediation of sites contaminated with hazardous wastes. Efforts to modify trees through biotechnology are in their infancy, and this review seeks to outline the underpinnings of what will undoubtedly be an area of increased emphasis in the next millennium. PMID- 9204751 TI - Microorganisms and enzymes involved in the degradation of plant fiber cell walls. AB - One of natures most important biological processes is the degradation of lignocellulosic materials to carbon dioxide, water and humic substances. This implies possibilities to use biotechnology in the pulp and paper industry and consequently, the use of microorganisms and their enzymes to replace or supplement chemical methods is gaining interest. This chapter describes the structure of wood and the main wood components, cellulose, hemicelluloses and lignins. The enzyme and enzyme mechanisms used by fungi and bacteria to modify and degrade these components are described in detail. Techniques for how to assay for these enzyme activities are also described. The possibilities for biotechnology in the pulp and paper industry and other fiber utilizing industries based on these enzymes are discussed. PMID- 9204752 TI - Hemicellulases in the bleaching of chemical pulps. AB - Hemicellulase-aided bleaching is the first full-scale biotechnical application in the pulp and paper industry which truly exploits the unique specificity and safety of biocatalysts. Hemicellulases are used to modify the structure of xylan and glucomannan in pulp fibers in order to enhance the chemical delignification. This technology can be combined with various types of kraft pulping processes and bleaching sequences. The aims of the enzymatic treatment depend on the actual mill conditions, and may be related to environmental demands, reduction of chemical costs, or maintenance or even improvement of product quality. The technology is applied on the mill scale in several countries. This review describes the principles of the enzyme-aided bleaching, the composition of the fiber substrates, the basic enzymology involved, and the present knowledge of the mechanisms of the action of enzymes, as well as the practical results and advantages obtained on the laboratory and industrial scale. PMID- 9204753 TI - Acid leaching studies of chrysotile asbestos from mines in the Coalinga region of California and from Quebec and British Columbia. AB - The dissolution of magnesium (Mg) and silicon (Si) from various samples of chrysotile asbestos was measured in N HCl at 25 degrees C. Nine samples were used, five from Canada and four from the Coalinga deposit in California. With milled samples from Quebec, the fraction of Mg dissolving was linearly related to the square root of the leaching time until at least 65% had dissolved. With a hand-picked sample of ore from Quebec, the sample from British Columbia and all the Californian samples, the Mg leaching patterns were sigmoid. The leaching patterns for Si were sigmoid in shape for all the materials tested. Mean Mg dissolution rates were calculated for each leaching period. Considerable differences were observed between samples from the different mining regions and also between hand-picked and milled samples from the same mine. Initially, Mg dissolved more rapidly from milled Quebec chrysotiles than from the Coalinga samples. This difference is due in part to the rapid dissolution of non structural brucite, present in all the samples from Quebec but not in those from California. An additional cause is greater damage to the fibre surfaces resulting from the milling to which the less readily-opened fibres, typical of the Quebec mining area, were subjected. Once this readily-available Mg had dissolved, there was little difference in leaching rates between milled and unmilled samples from the different regions. When the fraction of Mg dissolving is plotted against that of Si, all the materials follow a similar pattern, suggesting that the dissolution of Si (as silica) is the rate-controlling step in the dissolution of Mg. PMID- 9204754 TI - Acid leaching studies of neutron-irradiated chyrsotile asbestos. AB - Samples of chrysotile from Quebec (UICC B and Jeffrey 4T-30) and from the Coalinga region of California (Calidria RG-144) were irradiated with thermal neutrons in a reactor. The main activation products induced were 46Sc, 51Cr, 59Fe and 60Co. Accurately weighed samples of the irradiated materials were dispersed in N HCl by hand shaking for 10 s. After leaching for predetermined periods at 25 degrees C, the samples were filtered and the concentrations of Mg determined in the filtrates by inductively-coupled plasma atomic emission spectrometry (IPC AES) and the activities of the four radionuclides by high resolution gamma-ray spectrometry. Similar measurements were made on solutions obtained by refluxing samples of irradiated chrysotile with 2N HCl for 2 h. The specific activities of each of the four activation products were calculated in arbitrary units and, as the concentrations of Sc, Cr, Fe and Co in the UICC B sample had already been determined, it was possible to estimate the concentrations of these elements in the other two samples. Similarities in the leaching patterns of magnesium and of the activation products showed that with all samples, high proportions of the parent trace elements were present in the form of isomorphous substitutes for magnesium in structural brucite. Agreement was closest with the Calidria chrysotile in which all the radionuclides had a similar pattern. With the Jeffrey and UICC B samples, the presence of a high proportion of the iron as relatively insoluble magnetite accounted for the observed discrepancy in behaviour between 59Fe and Mg. More detailed calculation of leaching rates over specific time intervals showed that, initially, 51Cr and 60Co dissolved more rapidly than Mg but that this was followed by a period in which the opposite was the case. It was concluded that the Calidria RG-144 sample is an ideal candidate for studies of magnesium dissolution in vivo. PMID- 9204755 TI - Log-normality of distribution of occupational exposure concentrations to cobalt. AB - In evaluating occupational exposures, the daily exposure levels of a worker are generally assumed to be distributed log-normally between days. When there are many workers in a job group, the individual worker's arithmetic means are assumed to be distributed log-normally. Using the cobalt exposure data in a hard metal factory, these assumptions were examined with the Shapiro-Wilk W test. The hypothesis of log-normality was accepted in all cases, while the hypothesis of normality was rejected in some cases. PMID- 9204756 TI - Grouping strategies for exposure to inhalable dust, wheat allergens and alpha amylase allergens in bakeries. AB - This paper describes repeated measurements of inhalable flour dust, wheat allergens and alpha-amylase allergens in the bakery industry. A total of 571 full shift personal dust samples was collected. Wheat allergens and alpha-amylase allergens were measured in 449 and 507 samples, respectively, by the use of recently developed immunoassays. For all three measures of exposure, the main components of exposure variability were determined. Different grouping strategies for studying exposure-response relationships were compared. The specific job of a bakery worker was identified as the most important source of variability in inhalable flour dust concentrations. For exposure to wheat allergens, the job performed was also the most important source of variation, but type of bakery also explained some of the variability. For alpha-amylase allergen exposure, information on type of bakery was more important then job information. For exposure to inhalable dust and wheat allergens, a classification by job title would lead to sufficient contrast in average exposure levels. By contrast, a grouping strategy based on a combination of job and type of bakery appeared to be essential to obtain a useful classification of exposure to alpha-amylase allergens. 1997 British Occupational Hygiene Society. PMID- 9204757 TI - Dermal exposure of amateur or non-occupational users to wood-preservative fluids applied by brushing outdoors. AB - The results of an experiment to determine the likely dermal exposure of amateur or occasional users to wood-preservative fluids, applied by brushing onto a wooden fence outdoors are presented. Exposure was measured using FIVES, a fluorescence monitoring technique developed at the Health and Safety Laboratory. Dermal exposure could be examined and measured in a detail that would have been impossible using any other technique. A number of factors were found to affect dermal exposure, including some that were unexpected. Trousers, a long-sleeved shirt and permeable gloves offered 20 times the protection of shorts and a T shirt. Differences between individual subjects' behaviour gave rise to variations of a factor of 10. There was far more contamination at lower ambient temperatures, possibly because of more vigorous brushing. Spirit-based fluid caused more exposure than water-based fluid, probably because the spirit flicked easily from the brush as a spray whereas the water remained in soapy globules. Only 1.6 times more fluid was applied in 1 h than in 0.5 h, but it caused 3.7 times as much contamination. PMID- 9204758 TI - Accuracy and reproducibility of calibrations on the skin using the fives fluorescence monitor. AB - Dermal contamination from products such as pesticide formulations may be measured by adding a fluorescent dye as a tracer, and photographing the glow of the dye on the skin under ultraviolet light. The Fluorescence Interactive Video Exposure System (FIVES) fluorescence monitor, developed at the Health and Safety Laboratory, uses calibrations of intensity of glow against surface concentration of dye which are affected by the natural fluorescence of the skin and by skin hue. Calibrations of two dye/product mixtures upon the skin of several volunteers showed that a simple multiplicative correction procedure aligned all the responses to one calibration curve for each mixture. The palms of the hands were an exception, because their natural fluorescence was much higher than other areas of skin, and the relation no longer held. A further correction factor aligned their responses with the rest. The calibration data were fitted best by log-log polynomials, although linear polynomials were acceptable. Using this new procedure, the FIVES monitor could estimate dermal contamination to within +/- 70% of known applied doses. Upon exposure to light, the dyes on the skin were found to fade by 20% in 1/2 h, but no further in 1 h. The procedure and calibrations developed here were used in an experiment to estimate the likely dermal exposures of amateur or occasional biocide users, painting ready-to-use wood preservative fluids onto a trellis fence outdoors. PMID- 9204759 TI - Timing of sample collection for biological monitoring of occupational exposure. AB - The timing of sample collections for the biological monitoring of occupational exposure profoundly affects the resulting data. Sampling time with respect to the day in the working week and the end of exposure is crucial for measurements of rapidly excreted indicators of exposure. Owing to the cumulation of slowly excreted exposure indicators, timing of sample collection with respect to the duration of employment is essential. The steady state is established within a week, if the exposure indicator is excreted rapidly (with a half-life shorter than 45 h), or within months or years, if it is excreted slowly. In this study, exposure indicators are characterized by the elimination half-life. A monocompartmental model is used to calculate the biological levels at steady state and the duration of occupational exposure needed to reach the apparent steady state. PMID- 9204760 TI - A crystal milestone: the structure of regulated Src. AB - The viral and cellular forms of the Src protein tyrosine kinases take a prototypic role in oncology and signal transduction research, by virtue of being holders of an impressive number of 'firsts'. Our understanding of the biochemistry and physiology of Src has therefore always been used as a reference for our general advancement in the field of protein phosphorylation and growth control. The recent solution of the crystal structure of two members of the Src family represents a milestone in these disciplines and, as usual, provides a general lookout post for developments to come. PMID- 9204761 TI - Evolution of the multi-tubulin hypothesis. AB - Microtubules are organized into diverse cellular structures in multicellular organisms. How is such diversity generated? Although highly conserved overall, variable regions within alpha- and beta-tubulins show divergence from other alpha and beta-tubulins in the same species, but show conservation among different species. Such conservation raises the question of whether diversity in tubulin structure mediates diversity in microtubule organization. Recent studies probing the function of beta-tubulin isotypes in axonemes of insects suggest that tubulin structure, through interactions with extrinsic proteins, can direct the architecture and supramolecular organization of microtubules. PMID- 9204762 TI - A regulatory switch involving a Clp ATPase. AB - Clp ATPase chaperone proteins are found in procaryotes and eucaryotes. Recently, ClpC of Bacillus subtilis was found to be part of a regulatory switch(1). ClpC, in combination with the MecA and ComS proteins, regulates the activity of a transcription factor, ComK, which is necessary for the development of genetic competence (the ability to bind and take up exogenous DNA). The complex of ClpC:MecA:ComK renders ComK inactive. Interaction between ComS and the ternary complex releases active ComK. This regulatory switch controls ComK activity in response to cell density signals that affect production of ComS. Regulated interaction between Clp ATPase and target proteins might prove to be widespread. PMID- 9204763 TI - Genetic analysis of craniofacial development in the vertebrate embryo. AB - Every cartilage and bone in the vertebrate skeleton has a precise shape and position. The head skeleton develops in the embryo from the neural crest, which emigrates from the neural ectoderm and forms the skull and pharyngeal arches. Recent genetic data from mice and zebrafish suggest that cells in the pharyngeal segments are specified by positional information in at least two dimensions, Hox genes along the anterior-posterior axis and other homeobox genes along the dorsal ventral axis within a segment. Many zebrafish and human mutant phenotypes indicate that additional genes are required for the development of groups of adjacent pharyngeal arches and for patterning along the mediolateral axis of the skull. The complementary genetic approaches in humans, mice and fish reveal networks of genes that specify the complex morphology of the head skeleton along a relatively simple set of coordinates. PMID- 9204764 TI - PDZ domains: targeting signalling molecules to sub-membranous sites. AB - PDZ (also called DHR or GLGF) domains are found in diverse membrane-associated proteins including members of the MAGUK family of guanylate kinase homologues, several protein phosphatases and kinases, neuronal nitric oxide synthase, and several dystrophin-associated proteins, collectively known as syntrophins. Many PDZ domain-containing proteins appear to be localised to highly specialised submembranous sites, suggesting their participation in cellular junction formation, receptor or channel clustering, and intracellular signalling events. PDZ domains of several MAGUKs interact with the C-terminal polypeptides of a subset of NMDA receptor subunits and/or with Shaker-type K+ channels. Other PDZ domains have been shown to bind similar ligands of other transmembrane receptors. Recently, the crystal structures of PDZ domains, with and without ligand, have been determined. These demonstrate the mode of ligand-binding and the structural bases for sequence conservation among diverse PDZ domains. PMID- 9204765 TI - Patterning of the mammalian dentition in development and evolution. AB - The mammalian dentition is a segmented organ system with shape differences among its serially homologous elements (individual teeth). It is believed to have evolved from simpler precursors with greater similarities in shape among teeth, and a wealth of descriptive data exist on changes to the dentition that have occurred within mammals. Recent progress has been made in determining the genetic basis of the processes that form an individual tooth, but patterning of the dentition as a whole (i.e. the number, location and shape of the teeth) is less well understood. In contrast to similarly organized systems, such as the vertebral column and limb, Hox genes are not involved in specifying differences among elements. Nevertheless, recent work on a variety of systems is providing clues to the transcription factors and extracellular signalling molecules involved. PMID- 9204766 TI - The IRS-signalling system during insulin and cytokine action. AB - The discovery of the first intracellular substrate for insulin, IRS-1, redirected the field of diabetes research and has led to many important advances in our understanding of insulin action. Detailed analysis of IRS-1 demonstrates structure/function relationships for this modular docking molecule, including mechanisms of substrate recognition and signal propagation. Recent work has also identified other structurally similar molecules, including IRS-2, the Drosophila protein, DOS, and the Grb2-binding protein, Gab1, suggesting that this intracellular signalling strategy is conserved evolutionarily and is utilized by an expanding number of receptor systems. In fact, IRS-1 itself has been shown to be important in other growth factor and cytokine signalling systems, including growth hormone and several interleukins. Analysis of mice lacking IRS-1 confirms an important physiological role for this protein in glucose metabolism and general cell growth in the intact animal. Disregulation of the signalling pathways integrated by the IRS proteins may contribute to the pathophysiology of non-insulin-dependent diabetes mellitus or other diseases. PMID- 9204767 TI - Death substrates come alive. AB - Interleukin 1 beta-converting enzyme (ICE)-like proteases (caspases) play an important role in programmed cell death (apoptosis), and elucidating the consequences of their proteolytic activity is central to our understanding of the molecular mechanisms of cell death. Diverse structural and regulatory proteins and enzymes, including protein kinase C delta, the retinoblastoma protein (a protein involved in cell survival), the DNA repair enzyme DNA-dependent protein kinase and the nuclear lamins, undergo specific and limited endoproteolytic cleavage by various caspases during apoptosis. Since individual caspases can cleave multiple substrates, the consequences of cleavage of only a single substrate are still poorly understood. Nevertheless, proteolytic activation of protein kinase C delta may be an important early step in the cell death pathway, and cleavage of the retinoblastoma protein could suppress its cell survival function, whereas proteolytic inactivation of DNA repair enzymes might compromise the ability of the cell to reverse DNA fragmentation. On the other hand, cleavages of nuclear and cytoplasmic structural proteins (e.g. the lamins and Gas2) appear to be required for or contribute to the dramatic rearrangements in cellular architecture that are necessary for the completion of the cell death process. An emerging theme is that parallel and sequential proteolytic activation and inactivation of key protein substrates occurs during the multiple steps of apoptosis. PMID- 9204768 TI - Collectins: collectors of microorganisms for the innate immune system. AB - Collectins are a group of multimeric proteins mostly consisting of 9-18 polypeptides organised into either 'bundle-of-tulips' or 'X-like' overall structures. Each polypeptide contains a short N-terminal segment followed by a collagen-like sequence and then by a C-terminal lectin domain. A collectin molecule is assembled from identical or very similar polypeptides by disulphide bonds at the N-terminal segment, formation of triple helices in the collagen-like region and clusters of three lectin domains at the peripheral ends of triple helices. These proteins can bind to sugar residues on microorganisms via the peripheral lectin domains and subsequently interact, via the collagen-like triple helices, with receptor(s) on phagocytes and/or the complement system to bring about the killing and clearance of the targets without the involvement of antibodies. The collectins can also bind to phagocyte receptor(s) to enhance phagocytosis mediated by other phagocytic receptors. Lack, or low levels, of collectin expression can lead to higher susceptibility to infections, especially during childhood when specific immunity has not fully developed. Therefore, the collectins play important roles in the enhancement of innate immunity. PMID- 9204769 TI - Developmental phenotypic plasticity: where ecology and evolution meet molecular biology. AB - The plastic response of phenotypic traits to environmental change is a common research focus in several disciplines-from ecology and evolutionary biology to physiology and molecular genetics. The use of model systems such as the flowering plant Arabidopsis thaliana has facilitated a dialogue between developmental biologists asking how plasticity is controlled (proximate causes) and organismal biologists asking why plasticity exists (ultimate causes). Researchers studying ultimate causes and consequences are increasingly compelled to reject simplistic, 'black box' models, while those studying proximate causes and mechanisms are increasingly obliged to subject their interpretations to ecological 'reality checks.' We review the successful multidisciplinary efforts to understand the phytochrome-mediated shade-avoidance and light-seeking responses of flowering plants as a pertinent example of convergence between evolutionary and molecular biology. In this example, the two-way exchange between reductionist and holist camps has been essential to rapid and sustained progress. This should serve as a model for future collaborative efforts towards understanding the responses of organisms to their constantly changing environments. PMID- 9204770 TI - A hypothesis to explain why translation inhibitors stabilize mRNAs in mammalian cells: mRNA stability and mitosis. AB - Protein synthesis inhibitors prolong the half-lives of most mRNAs at least fourfold in the somatic cells of higher eukaryotes and in yeast cells. Some mRNAs are stabilized because the inhibitors affect mRNA-specific regulatory factors; however, hundreds or thousands of other mRNAs are probably stabilized by a common mechanism. We propose that mRNA stabilization in cells treated with a translation inhibitor reflects a physiological process that occurs during each mitosis and is important for cell survival. Transcription and translation rates decline drastically during a 1-2 hour interval of mitosis. We hypothesize that translational repression during this interval somehow inactivates a critical component of the mRNA degradation machinery. As a result, mRNA half-lives are prolonged during the interval when transcription is repressed. If labile mRNAs were not stabilized during mitosis they, and perhaps also the labile proteins they encode, would be depleted as the cell entered G1 phase, with deleterious consequences. Stabilization during mitosis, or in response to translation inhibitors, thus preserves the capacity of the cell to synthesize essential proteins as it enters G1 or recovers from inhibitor treatment. mRNA stabilization might serve a similar purpose during starvation or any stress negatively affecting translation. PMID- 9204771 TI - The risk society in an age of anxiety: situating fear of crime. AB - As many now recognize, fear of crime is an inadequately theorized concept. In particular, it is premissed on rational, calculating individuals who routinely miscalculate their 'true' risk of crime. Hence the repeatedly found paradox that the least at risk group (elderly females) are most fearful. The risk literature has adopted a cultural/anthropological rather than an individual perspective, but, in so doing has not succeeded in retheorizing the notion of the rationally calculating subject it critiques (Douglas), even if rational calculations are no longer possible in today's 'risk society' (Beck). We develop these cultural perspectives in a way which is founded on a post-structuralist theory of individuals wherein inter-subjective defending against anxiety replaces rational calculation as central to the understanding of fear. Not only does this re-link the concepts of fear and anxiety, currently divorced in the fear of crime debate, but it offers the prospect of understanding the paradoxical mismatch between risk and fear at both the level of the individual and of society. PMID- 9204772 TI - Oligonucleotide-directed mutagenesis and subsequent expression of the corresponding recombinant proteins without changing the bacterial vector system. AB - A new bacterial vector was constructed that combines the attractive features of gene fusion vectors and phagemids. A gene of interest cloned into this new vector can either be expressed as fusion protein or be prepared as single-stranded template DNA within the same system. Thus, time consuming subcloning procedures changing the bacterial vector according to the required method are avoided. As an example sequencing, expression and subsequent purification of site-directed mutants of herpes simplex virus type 1 thymidine kinase are discussed. PMID- 9204773 TI - Inhibition of benzodiazepine binding in vitro by amentoflavone, a constituent of various species of Hypericum. AB - Flower extracts of Hypericum perforatum, Hypericum hirsutum, Hypericum patulum and Hypericum olympicum efficiently inhibited binding of [3H]flumazenil to rat brain benzodiazepine binding sites of the GABAA-receptor in vitro with IC50 values of 6.83, 6.97, 13.2 and 6.14 micrograms/ml, respectively. Single constituents of the extracts like hypericin, the flavones quercetin and luteolin, the glycosylated flavonoides rutin, hyperoside and quercitrin and the biflavone 13, II8-biapigenin did not inhibit binding up to concentrations of 1 microM. In contrast, amentoflavone revealed an IC50 = 14.9 +/- 1.9 nM on benzodiazepine binding in vitro. Comparative HPLC analyses of hypericin and amentoflavone in extracts of different Hypericum species revealed a possible correlation between the amentoflavone concentration and the inhibition of flumazenil binding. For hypericin no such correlation was observed. Our experimental data demonstrate that amentoflavone, in contrast to hypericin, presents a very active compound with regard to the inhibition of [3H]-flumazenil binding in vitro and thus might be involved in the antidepressant effects of Hypericum perforatum extracts. PMID- 9204774 TI - Comparative study of in-vitro release and bioavailability of sustained release diclofenac sodium from certain hydrophilic polymers and commercial tablets in beagle dogs. AB - Hydrophilic colloids interact with metallic ions to yield crosslinked insoluble salts. Such principle was utilized in the preparation of diclofenac sodium beads from sodium alginate and sodium carboxymethylcellulose. Hard spherical beads of aluminum alginate and aluminum carboxymethylcellulose with a narrow particle size distribution (1.60 +/- 0.12 and 3.10 +/- 0.20 mm) and low friability (0.5 and 1.4%) respectively were obtained with high yield (80-90%) and a drug content approaching 70-80%. The type and concentration of the polymers as well as the pH of the dissolution medium were found to affect the rate of drug release. Beads prepared from Na-alginate showed a non-significantly (p > 0.05) faster rate of drug release than that prepared from NaCMC. The higher the polymer concentration, the slower was the rate of drug release. Diclofenac sodium did not release in 0.1 N HCl (pH 1.2) for 2 h and released in phosphate buffer solution (pH 6.8) from the two formulations studied and from the commercial Voltaren Retard tablet. The two formulations of the beads resulted in a sustained release action of diclofenac sodium for 24 h. They showed Kel values of 0.02 +/- 0.01 and 0.3 +/- 0.01 h-1 and these correspond to t1/2 of 34.65 and 27.70 for the Na-alginate and NaCMC beads, respectively. They also showed mean residence time (MRT) values of 9.56 +/- 2.5 and 7.86 +/- 0.54 h, respectively. They also showed non-significant (p > 0.05) differences with respect to their plasma levels, Cmax, Tmax and AUC0- >24 h. The relative bioavailability of the two formulations were 59.01 and 47.96%, respectively, relative to that of the commercial Voltaren Retard tablets of Ciba-Geigy which showed a Kel of 0.044 h-1 corresponding to a t1/2 of 15.75 h and MRT of 7.45 +/- 1.10 h. PMID- 9204775 TI - In vitro characterization of methotrexate loaded poly(lactic-co-glycolic) acid microspheres and antitumor efficacy in Sarcoma-180 mice bearing tumor. AB - Methotrexate (MTX) loaded poly (lactic-co-glycolic) acid (PLGA) microspheres were prepared by emulsion solvent evaporation technique. The mean diameter of the microspheres was affected by the type of emulsion stabilizer, polymer concentration, aqueous and organic phase volume and stirring speed. The in vitro release was triphasic and was dependent on copolymer composition and molecular weight of the polymer. Antitumor efficacy in Sarcoma-180 tumor bearing mice exhibited increased volume doubling time (18 +/- 2.7 days) compared to plain subcutaneous injection of methotrexate (8 +/- 0.7 days). Preliminary pharmacokinetic studies following subcutaneous administration of MTX loaded PLGA microspheres illustrated the controlled release of the drug. The studies demonstrated the feasibility of employing PLGA as an effective carrier for antineoplastic drug like methotrexate. PMID- 9204776 TI - Determination of psilocin and 4-hydroxyindole-3-acetic acid in plasma by HPLC-ECD and pharmacokinetic profiles of oral and intravenous psilocybin in man. AB - In order to investigate the pharmacokinetic properties of psilocybin (PY), the main psychoactive compound of Psilocybe mushrooms, high performance liquid chromatographic procedures with column-switching coupled with electrochemical detection (HPLC-ECD) for reliable quantitative determination of the PY metabolites psilocin (PI) and 4-hydroxyindole-3-acetic acid (4HIAA) in human plasma were established. Sample work-up includes protection of the highly unstable phenolic analytes with ascorbic acid, freeze-drying and in-vitro microdialysis. The data of two controlled clinical studies with healthy volunteers are presented. The subjects (N = 6 for both studies) received single oral PY doses of 0.224 +/- 0.02 mg/kg b.wt. (10-20 mg) and intravenous doses of 1 mg PY, respectively. Peak plasma levels of PI after oral administration of PY were measured after 105 +/- 37 min showing an average concentration of 8.2 +/- 2.8 ng PI/ml plasma. 4HIAA peak concentrations of 150 +/- 61 ng/ml plasma were found 113 +/- 41 min after ingestion of PY. After intravenous administration, a mean PI maximum plasma concentration of 12.9 +/- 5.6 ng/ml plasma was found 1.9 +/- 1.0 min after injection. The maximum plasma levels appearing within a very short period indicate a rapid dephosphorylation of PY also when administered systemically. 4HIAA was not detected after 1 mg of intravenous PY. Estimates for the absolute bioavailability of PI after oral administration of PY were 52.7 +/- 20% (N = 3). PMID- 9204777 TI - Models of the self: self-construals and gender. AB - The authors first describe individual differences in the structure of the self. In the independent self-construal, representations of others are separate from the self. In the interdependent self-construal, others are considered part of the self (H. Markus & S. Kitayama, 1991). In general, men in the United States are thought to construct and maintain an independent self-construal, whereas women are thought to construct and maintain an interdependent self-construal. The authors review the psychological literature to demonstrate that many gender differences in cognition, motivation, emotion, and social behavior may be explained in terms of men's and women's different self-construals. Recognition of the interdependent self-construal as a possible alternative conception of the self may stimulate new investigations into the ways the self influences a person's thinking, feeling, and behaving. PMID- 9204778 TI - What do men want? Gender differences and two spheres of belongingness: comment on Cross and Madson (1997) AB - In response to S. E. Cross and L. Madson's (1997) suggestion that men's behaviors reflect a desire for independence and separateness, the authors propose that those same behaviors are designed to form connections with other people but in a broader social sphere. Women's sociality is oriented toward dyadic close relationships, whereas men's sociality is oriented toward a larger group. Gender differences in aggression, helping behavior, desire for power, uniqueness, self representations, interpersonal behavior, and intimacy fit this view. PMID- 9204779 TI - A developmental perspective of self-construals and sex differences: comment on Cross and Madson (1997) AB - S. E. Cross and L. Madson (1997) proposed that women and men differ in self construals, with women as interdependent and men as independent, and that these construals are seen to underlie many sex differences in social behavior. In this article, the authors address the issues of sex differences in self-construals, the stability of self-construals, and the centrality of interdependence independence to sex differences. They examine the proposal next from a developmental perspective, suggesting that development of the self does not precede children's gender-related behavior. Evidence regarding the socialization processes that may contribute to sex differences in self-construals is largely inconclusive, especially regarding the differential treatment of boys and girls by parents. Their developmental perspective provides insights into ways that Cross and Madson's approach can be elaborated to explain sex differences in social behavior. PMID- 9204782 TI - Imagery in human classical conditioning. AB - Many clinical strategies use patients' imagery to explore and treat phobic and posttrauma reactions, however little attention has been paid to the underlying assumption that imagery of relevant stimuli may help maintain conditioned behavior. In this article, the authors examine the premise that mental images can potentiate and substitute for physical stimuli in human classical conditioning. The authors review empirical evidence to detail the role of images of conditioned stimuli (CS) and unconditioned stimuli (US) during pre-exposure to stimuli, the actual pairing of the CS and US, and extinction when the CS is presented alone. The evidence suggests that mental imagery can facilitate or diminish the outcome of classical conditioning in humans and, more tentatively, that mental images can substitute for actual US and CS in autonomic conditioning. They argue that researchers should explore the role of mental imagery in conditioning through the use of advances in the measurement of imagery. Finally, they analyze anxiety and trauma reactions as examples of how applied areas can be used to explore and benefit from developments in this area. PMID- 9204781 TI - Is alcohol a cofactor of HIV and AIDS? Evidence from immunological and behavioral studies. AB - The authors aim to critically examine empirical research on the effects of alcohol on HIV and AIDS from the immunological and behavioral fields. In vitro immunological studies demonstrate that social drinking increases the susceptibility of human cells to HIV infection. Animal studies show that acute and chronic alcohol ingestion increases rate of progression from retrovirus to clinical illness. In humans with HIV, no experimental evidence shows that alcohol is a cofactor of AIDS. Findings from behavioral studies show that a link between social drinking and risk of HIV is weak. No experimental evidence demonstrates that chronic drinking influences rate and course of disease progression to AIDS in humans who are HIV+. It is premature to promote the role of alcohol as a cofactor in HIV and AIDS. PMID- 9204783 TI - Revised antitrust policy statements on healthcare. PMID- 9204784 TI - Expert testimony. PMID- 9204785 TI - Interprofessional jousting and medical tragedies: strategies for enhancing professional relations. AB - In 1990, the AANA Council for Public Interest in Anesthesia (CPIA) in conjunction with Anesthesia Professional Liability Services Inc. and the St. Paul Fire and Marine Insurance Company surveyed AANA members to identify stressors that have the most impact on performance, health, and the risk of a lawsuit. The results were presented that year in the AANA Annual Meeting's keynote address. Among the most significant findings was dysfunctional CRNA/physician relationships. In light of this finding, the CPIA and The St. Paul developed and sponsored a one day enhancing professional relations workshop which has been offered at the past five annual meetings. This article summarizes the activities and key learnings from that workshop. PMID- 9204786 TI - Implications of recent antiviral pharmacology to prevention of occupational HIV transmission in anesthesia clinical practice. PMID- 9204787 TI - Anesthesia providers, patient outcomes, and costs: a critique. AB - The June 1996 article in Anesthesia and Analgesia by Abenstein and Warner entitled "Anesthesia Providers, Patient Outcomes, and Costs" presents important information about anesthesia services, but it contains a number of errors and questionable interpretations that could lead to inappropriate programs and policies. Among the most important points of fact we clarify in our paper are: 1. Three organizations that accredit, certify, and govern nurse anesthetists are organized in similar fashion to three comparable bodies governing anesthesiologists. There is no justification for the implication that the AANA somehow controls the accreditation and certification of CRNAs. 2. The conclusion that anesthesiologistled care teams are the preferred model for all anesthesia services and settings because of improved patient outcomes is overly simplistic and is not borne out in the literature. 3. The attribution of reduced mortality from anesthesia over the past 40 years to the increase in numbers of anesthesiologists is not justified. Many other factors, including new anesthetic agents and improved patient monitoring, also are important. 4. The use of a hypothetical example related to Medicare reimbursement in New York to justify the implication that CRNA-delivered services are more costly than anesthesiologist delivered services is misleading and not borne out in the literature. We hope that planners and policy makers will read the article by Abenstein and Warner with extreme caution. Taking some of their statements and conclusions seriously could lead to policies and programs that are not focused in science. PMID- 9204788 TI - Effect of preemptive acetaminophen on postoperative pain scores and oral fluid intake in pediatric tonsillectomy patients. AB - Postoperative pain is a significant problem that continues to be undertreated in the pediatric population. Preemptive administration of analgesics has recently emerged as a method to enhance pain management associated with surgery. The purpose of this study was to compare postoperative pain scores, rescue analgesic use, and oral fluid intake in children who received acetaminophen preoperatively to children who received postoperative acetaminophen. The sample consisted of 28 children, 2-8 years of age, scheduled for elective tonsillectomy. Children were randomized into the control or the experimental groups. Anesthesia induction and maintenance were standardized. The experimental group received 15 mg/kg of oral acetaminophen preoperatively, and the control group received 20 mg/kg of rectal acetaminophen postoperatively. Pain was scored with the FLACC (faces, legs, activity, cry, consolability) behavioral assessment tool. Scores were significantly lower for the experimental group at 30 minutes after awakening and significantly lower for the control group at 240 minutes (P < .05). Eight patients (57%) in the control group and only 4 (28%) in the experimental group required rescue morphine postoperatively. Total postoperative morphine was not significantly different between groups. There were no differences in time to initial oral fluid intake and total oral fluid intake postoperatively. Incidence of nausea and vomiting was high in both groups (64-78%). These results provide evidence that preemptive acetaminophen may enhance analgesia in pediatric tonsillectomy patients. Preoperative acetaminophen is a safe, quick, and inexpensive intervention that can readily be incorporated into anesthesia practice. PMID- 9204789 TI - Anesthetic considerations for the new antiobesity medications. AB - Combination drug therapy can effectively treat the problem of obesity. The most commonly used combination is a mix of fenfluramine and phentermine. Fenfluramine inhibits the reuptake of serotonin and acts on the hypothalmic appetite control center, while phentermine acts as an appetite suppressant. These drugs along with diet and exercise effectively help people to lose weight with few side effects. However, there are several anesthetic considerations when providing anesthesia services for patients on the fenfluramine and phentermine regimen. Problems of hypotension on induction, hypoglycemia, hyperthermia, and pulmonary hypertension have been reported in the literature. Recently, dexfenfluramine (Redux) was approved by the U.S. Food and Drug Administration. It is the dextrostereoisomer of fenfluramine and is believed to produce the same weight loss with less side effects. Anesthesia providers must understand the potential risks involved when administering a general anesthetic to these patients. PMID- 9204790 TI - The effect of glycopyrrolate premedication on postoperative sore throat. AB - Patients given general endotracheal anesthesia commonly experience postoperative sore throat and/or hoarseness. Our study examined whether the occurrence of postoperative sore throat was associated with the use of a glycopyrrolate premedication and found that it was. We randomly assigned 120 patients undergoing general endotracheal anesthesia for routine surgery to receive a preoperative mediation including or excluding glycopyrrolate. We controlled for factors known to increase the risk of a postoperative sore throat. After surgery, an interviewer, unaware of the subject's group assignment, questioned each subject about the presence of a sore throat and, if present, asked the patient to rate its severity. We found that patients who did not receive preoperative glycopyrrolate were significantly less likely to report having a sore throat or reported having a less severe sore throat than patients who did receive glycopyrrolate. PMID- 9204792 TI - Council on accreditation of nurse anesthesia educational programs. PMID- 9204791 TI - AANA Journal course: update for nurse anesthetists--genetic testing for malignant hyperthermia. AB - The "gold standard" for the determination of susceptibility to malignant hyperthermia has long been the caffeine-halothane-contracture test, which is costly and invasive. As the workings of molecular genetics are better understood, research is being applied to finding the causative gene for malignant hyperthermia. Once this gene is identified, genetic testing will involve a much simpler, less invasive test that uses blood samples to detect susceptibility. To reach this goal, researchers have been attempting to identify the deoxyribonucleic acid (DNA) mutations responsible for malignant hyperthermia by using techniques such as linkage analysis. Review of the literature reveals that malignant hyperthermia has been linked to the ryanodine receptor in swine and in some humans. It has also been linked to chromosome 19q12-13.1, which is where the gene encoding the ryanodine receptor lies. The literature further reveals that malignant hyperthermia may be a heterogeneous disorder, which means that more than one gene is responsible for its expression. PMID- 9204793 TI - Mack: an autobiography. PMID- 9204794 TI - Chair design and the anesthesia provider. PMID- 9204795 TI - The sesquicentennial of other anesthesia. PMID- 9204796 TI - The change leader. PMID- 9204799 TI - Measuring prevalence and incidence of pressure ulcers. AB - The current competitive health care environment has intensified the need for data that provide a snapshot of the realities of clinical practice. As decision making moves from a clinically based perspective to one grounded in scientific data, health care providers are increasingly being challenged to document the extent of a problem and the effectiveness of its management. This is especially true with pressure ulcers, which are viewed as high-volume, high-risk problems in most health care settings. Moreover, in long-term care facilities, regulatory agencies have designated the development of pressure ulcers as an indicator of quality of care provided to patients. Thus, it is essential that data related to the scope and severity of pressure ulcers in a facility be gathered accurately. The aim of this article is to describe a methodology for determining prevalence and incidence of pressure ulcers that accurately measures the effectiveness of preventive intervention. The importance of risk assessment and of clear operational definitions of the population and a case will be addressed. PMID- 9204800 TI - An alternative method for reducing plantar pressures in neuropathic ulcers. AB - Neuropathic foot ulcers, a serious complication of impaired pedal sensation, are the most common complication of diabetes mellitus leading to hospitalization. One of the goals in treating neuropathic ulcers is to reduce or eliminate pressure from the ulcer site to prevent further breakdown and allow healing. These treatments must address several forms of stress-vertical pressure, shearing force, and tissue strain. This paper reviews several techniques for off-weighting plantar ulcers and describes and recommends the felt-to-foam contact padding technique used at the Northern California Center for the Diabetic and Insensitive Foot and the California College of Podiatric Medicine. PMID- 9204801 TI - Nutritional supplements in the treatment of pressure ulcers: practical perspectives. AB - Because a pressure ulcer may be related to malnutrition, clinicians should assess patients' nutritional status and ensure that patients receive adequate nutrition to support healing. The Agency for Health Care Policy and Research's guideline Treatment of Pressure Ulcers outlines recommended calorie and protein levels for patients at risk for pressure ulcers and with pressure ulcers. This article focuses on practical aspects of providing adequate nutrition to patients with pressure ulcers, using a variety of products available for oral supplementation. PMID- 9204802 TI - A 4-year outcome-based retrospective study of wound healing and limb salvage in patients with chronic wounds. AB - This multicenter, retrospective study evaluated the wound healing and limb salvage outcomes over a 4-year period in 3,830 patients in 39 hospital-affiliated Wound Care Centers. These centers provide comprehensive outpatient wound care for chronic, nonhealing wounds. Two distinct outcomes were identified: (1) wound healing with comprehensive wound care (CWC) alone, and (2) wound healing with comprehensive wound care plus platelet releasate (CWC+PR). Data were analyzed with respect to healing and limb salvage in two groups: 1,019 patients who received CWC and 2,811 patients who received CWC+PR. Analysis of the standardized, customized database showed that overall healing rates were higher (p < .00001) and amputation rates were lower (p = .00005) in the CWC+PR group than in the CWC group. In addition, when healing rates were analyzed according to underlying condition, patients with all underlying conditions except autoimmune disorders showed higher healing rates in the CWC+PR group than in the CWC group. This study showed that patients treated with comprehensive wound care plus topical use of autologous platelet releasate had significantly higher rates of wound healing and increased limb salvage for most wounds than those treated with comprehensive wound care alone. PMID- 9204804 TI - Venous leg ulcer risk factors: who will study the problem? PMID- 9204803 TI - A comparison of two pressure-relieving devices on the prevention of heel pressure ulcers. AB - The effectiveness of hospital pillows versus a commercial heel elevation device (the Foot Waffle [EHOB incorporated]) in preventing heel pressure ulcers was examined using an experimental balanced factorial design with repeated measures on 52 patients (ages 27 to 90) in randomized groups. Heel interface pressures were taken with patients in supine and right lateral tilt positions. Logistic regression demonstrated a statistically significant difference between interface pressures on left and right heels (p = .004) and a trend toward significance between the pillow and Foot Waffle (p = .069). The Generalized Estimating Equations (GEE) method revealed the Foot Waffle was four times more likely not to suspend the heel off the bed than the pillow, and the left heel was four-and-a half times more likely to have higher interface pressures than the right. There was no significant difference between groups in incidence of lower-extremity pressure ulcers, but patients using the Foot Waffle developed pressure ulcers significantly sooner (10 days versus 13 days for the pillow). Heels require additional protection beyond the use of specially beds and mattress overlays. In order to provide continuous heel suspension, clinicians must consider proper fit, turning schedules, patient position, patient activity, and presence of additional equipment when selecting heel protection products. This study illustrates how difficult it is to control for all these factors when doing clinical research. Note: This study was done with a Foot Waffle model that has since been redesigned. No research is available on the new model. PMID- 9204805 TI - Comparison of two pressure-relieving devices on the prevention of heel pressure ulcers. PMID- 9204807 TI - The future: society and government working together on wound care. PMID- 9204806 TI - A foot "ulcer" resistant to healing. Acral-lentiginous melanoma. PMID- 9204808 TI - Diabetic foot wounds: pathogenesis and management. AB - The diabetic foot is prone to foot ulceration, which may lead to ischemia, infection, and the need for amputation. To try to reduce foot ulcer-related hospitalizations and decrease the amputation rate, health care providers need to understand the pathophysiology of the diabetic ulcer, treat ulcers promptly and aggressively, provide revascularization when necessary, prescribe therapeutic shoes, use a team approach, and conduct an intensive, ongoing patient-education program in foot care. PMID- 9204809 TI - Debridement: choices and challenges. AB - Debridement of nonviable tissue is crucial to optimal wound healing, which can be impaired unless all necrotic tissue, exudate, and metabolic wastes have been removed from the wound. Debridement methods are classified as sharp, mechanical, chemical, and autolytic. This article describes methods of debridement and their outcomes. PMID- 9204810 TI - Management of the acute burn wound: an overview. AB - Goals for managing an acute burn wound are similar to those of other wounds such that infection and scar formation are minimized, a moist wound environment is provided, and the surrounding tissue is protected from trauma. A variety of cleansing techniques are used with burn wounds, including local wound care and nonsubmersion and immersion hydrotherapy. Topical agents have significantly decreased the development of burn wound sepsis since the 1960s, and now various experimental agents are being investigated to improve wound healing. The choice of dressings depends on many patient and wound-related factors, and synthetic, biologic, and biosynthetic dressings are used to treat the different depths of burn wounds. However, skin grafts and the newer cultured skin substitutes remain the mainstay for healing a full-thickness burn wound. PMID- 9204811 TI - State and association/certifying boards CE requirements. PMID- 9204812 TI - Coproviding continuing education through faculty practice: a win-win opportunity. AB - Practice incorporated as an integral component of the faculty role benefits faculty, their clients, students, the school of nursing, and the community. When practice includes education, particular benefits result for those who have limited access to continuing nursing education (CNE). This article describes one school's way of responding to these learning needs. Three essential elements to a successful venture are described: a) a need for CNE is identified, b) faculty with diverse expertise are available, and c) a working relationship exists between an accredited CNE provider and the faculty. Other schools of nursing are encouraged to develop similar programs. PMID- 9204813 TI - The North Carolina AHEC Self-Paced RN Refresher Program: an evaluation of the first two years. AB - The North Carolina Area Health Education Centers (AHEC) Self-Paced RN Refresher Program was designed to promote reentry into practice for inactive nurses. The program consists of a didactic self-study correspondence course and a clinical practicum whereby the refresher nurse works one-on-one with a nurse preceptor. At the end of the first two years, the program was evaluated to determine its effectiveness in terms of participant satisfaction and return to nursing practice and to describe the nurse refresher population demographically. Participants reported a high degree of satisfaction; 69% of nurses who completed the program returned to nursing practice. The typical refresher student is a female Caucasian in her forties, married, and graduated from a diploma nursing program. PMID- 9204814 TI - New graduates' perceptions of clinical practice. AB - This study describes the stresses and challenges experienced by graduate nurses in clinical practice during their initial orientation period and examines the relationship of social support to these stresses. Thirty-five graduate nurses completed a modified Pagana Clinical Stress Questionnaire and social support measure during their orientation period. The graduates experienced a moderate degree of stress in their orientation. Stresses identified most frequently were: lack of experience as a nurse, interactions with physicians, lack of organizational skills and new situations and procedures. Although orientation was stressful, when describing emotions experienced during this period, graduates reported positive emotions most frequently. Pearson correlation revealed no significant relationship between social support and stress. Significant correlations were found, however, between social support and stimulation in clinical practice (r = 57, p = .001) and development of self-confidence (r = 39, p = .029). Findings highlighted the important role of the preceptor during orientation. PMID- 9204815 TI - Nurses and Public Law 102-119: a family-centered continuing education program. AB - The passage of Part H of the Education of the Handicapped Act, Public Law 99-457, and the reauthorization of the law in 1991, as Public Law 102-119, mandates a family-centered approach to the provision of services to infants and young children with handicaps. Nurses are named as providers in this federal legislation. The authors conducted a survey to assess the learning needs of nurses who may wish to serve as providers under the law. The survey results were used to develop a family-centered continuing education curriculum that pairs parents and nurse faculty as partners in delivering the curriculum. PMID- 9204816 TI - Returning to school: ten considerations in choosing a BSN program. AB - Ten factors, accreditation, cost, flexibility, location, method of earning credit, student profile, quality, progression to master's degree, resources and time, should be investigated before selecting a baccalaureate nursing program. A sample of 361 RNs enrolled in ten baccalaureate nursing programs identified and prioritized the ten factors. The most important factor was cost, followed by accreditation and flexibility. Asking questions in each of these areas enables the RN to make an informed decision before investing time, energy and money in a specific program. These ten factors can be guidelines used for assessing various program objectives. The subjective data are how each RN prioritizes the ten factors. PMID- 9204817 TI - The enhancement of writing skills among post-RN students. AB - Nurses need to communicate in oral and written formats, but the emphasis on concise charting and multiple-choice examinations hinders the development of scholarly writing skills. A questionnaire completed by 97 students identified the areas within writing and formatting that needed strengthening. Three strategies, useful to continuing education programs for nurses, were incorporated in a post RN program to enhance the students' writing abilities: a writing skills workshop, an in-class writing exercise and a draft paper assignment. Student evaluations have supported these strategies as well as guidance by faculty in assisting students to improve their writing. PMID- 9204818 TI - Continuing education? No problem. PMID- 9204819 TI - The cultural competent challenges in providing human services. PMID- 9204820 TI - Utilization of curanderos among foreign born Mexican-American women attending migrant health clinics. AB - This study explores the parallel use of "folk healers" and modern medicine among foreign born, Mexican-American women attending migrant health clinics in rural, eastern Washington state. Face-to-face interviews (n = 434) revealed that 21.4% of the women had sought care from curanderos within the past five years. Statistically significant predictors of utilization included Spanish as the language of preference (odds ratio = 2.58), having resided in the U.S. from one to five years (odds ratio = 2.82), and having received medicine or medical care from Mexico within the prior five years (odds ratio = 9.22). Implications for providers working in cross-cultural settings are discussed. PMID- 9204822 TI - Assessment of level of comfort in providing multicultural nursing care by baccalaureate nursing students. AB - The purpose of this study was to measure the level of comfort in providing transcultural nursing care to clients of diverse cultural backgrounds by graduating baccalaureate students. The sample consisted of 71 senior baccalaureate students. The Cultural Self-Efficacy Scale developed by Bernal and Froman was modified and used to measure the confidence level of the graduating nurses in providing transcultural care to five major minority groups in Michigan. The Cultural Self Efficacy Scale is a 26-item Likert type scale which is grouped into two categories: knowledge of cultural concepts for specific ethnic groups (African Americans, Latino-Hispanics, Middle Easterners/Arabics, Asian/Pacific Islanders, and Native American), and confidence in transcultural nursing skills. The items are rated on a scale of 1-5. Results of the study indicate a total mean cultural self-efficacy rating of 2.6. This suggests that the graduating students had little self confidence in their ability to give culturally congruent care. This was evident across the five ethnic groups. Test item analysis for specific ethnic groups suggest that students felt the least degree of confidence in their knowledge of class structure and migration patterns. They felt the greatest degree of confidence in their knowledge of family organization and economic style of living. Test item analysis for the skills suggest that students had the highest degree of confidence in their ability to use interpreters that were provided by either the family or the hospital. They had the least amount of confidence in their ability to instruct clients in child care and evaluating the effectiveness of discharge teaching. Results suggest that the sample of graduating baccalaureate students studied do not feel confident about caring for Michigan's five major ethnic groups. The data obtained in this study support previous research that nursing students are not provided with the experiences needed to give transcultural care to ethnically diverse populations. PMID- 9204821 TI - Cultural diversity process improves organizational community in urban teaching medical center. AB - An urban teaching facility with nearly 3,000 employees had communication problems associated with race, gender and other cultural differences. It also competed for health care dollars and faced possible reduction in federal funding. The medical center instituted mandatory training in cultural diversity and customer service and integrated the training process with the hospital's overall quality improvement plan and marketing strategy. The integrated approach affected the bottom line-Hurley's patient base has increased, and the medical center operates in the black. Training in cultural diversity and customer service is an effective tool to improve employee communication and improve financial outlook. PMID- 9204823 TI - African American children and adolescents exposure to community violence: a pilot study. AB - Children and adolescents living in communities with frequent episodes of violence are an at-risk population. Chronic exposure to community violence can compromise children and adolescents' health, cognitive functioning, and development. This interdisciplinary pilot study was conducted in an urban community to ascertain a description of children and adolescents' exposure to the type and frequency of violence. Children were found to witness more community violence, while adolescents both witness and experience more community and individual-level violence. Reduction of violence and creation of healthy and safe communities are critical to the well-being and productivity of all residents. PMID- 9204824 TI - Meeting the challenge of culture care in nursing: diversity, sensitivity, competence, and congruence. AB - Using examples from a number of developed and developing countries, this article defines, illustrates, compares, and contracts cultural diversity, cultural sensitivity, and cultural competence. Leininger's Sunrise Model provides the theoretical basis for determining nursing care that is culturally congruent. Two simulation games, designed to enhance cultural sensitivity, are included as well as a glossary of terms. PMID- 9204825 TI - Assessment of the health needs of low income, inner city, African American elderly. PMID- 9204826 TI - Past successes and future hopes in ophthalmology. PMID- 9204827 TI - How does vision affect learning? PMID- 9204828 TI - Visual consequences of ocular and adnexal findings in patients with Goldenhar's syndrome. PMID- 9204829 TI - Use of perfluorocarbon liquid in the repair of retinoschisis retinal detachments. PMID- 9204831 TI - Eyes on the Web. PMID- 9204832 TI - Identifying and managing complications in soft lens wearers. PMID- 9204833 TI - One thousand cataract patients on a Monday? PMID- 9204834 TI - The foundation of excellent surgical assisting. PMID- 9204835 TI - Fitting the keratoconic cornea. PMID- 9204836 TI - Comparison of the HOTV and Lea Symbols charts for preschool vision screening. PMID- 9204838 TI - Self-assessment. Molluscum contagiosum. PMID- 9204837 TI - The Roy adaptation model and its application to clinical nursing practice. PMID- 9204839 TI - Identifying and managing complications in soft lens wearers: Part II. PMID- 9204842 TI - Using technology to enhance information sharing. PMID- 9204840 TI - Corneal topography to help detect keratoconus. PMID- 9204843 TI - The transition from radio to television and its implications for WOC nursing. PMID- 9204844 TI - Connected to the future. PMID- 9204845 TI - Myra E. Levine: a theoretic basis for ET nursing. AB - ET nursing is a challenging specialty that requires exact knowledge, complex skills, and a holistic approach to patients with wound, ostomy and continence problems. Myra E. Levine's conceptual framework for nursing practice is described and applied to ET specialty practice nursing. Examples of Levine's four conservation principles-energy, structure, personal integrity, and social integrity-are integrated into the nursing care of patients' wound, ostomy, and continence problems. PMID- 9204846 TI - Sterile versus clean technique in postoperative wound care of patients with open surgical wounds: a pilot study. AB - PURPOSE: This study was completed to determine whether there were differences between sterile versus clean dressing change technique for open surgical wounds in the postoperative period with respect to (1) rate of wound healing and (2) cost of supplies. METHODS: A two-group design was used for this pilot study. Of a sample of 30 patients undergoing elective gastrointestinal operations with wounds healing by secondary intention, 15 were men and 15 were women. Mean age was 40.6 years (SD 13.0 years). Patients were randomly assigned to receive clean or sterile dressings, and the intervention was begun on the first postoperative day and repeated three times a day until discharge from the hospital. Analysis of rate of healing was performed with the Mann-Whitney U test: cost analysis was completed with a t test. FINDINGS: Subjects were studied for 3 to 9 days. Groups were homogeneous of the start of treatment with respect to age, length of operation, wound volume, nutritional status, and perfusion. There was no difference in rate of wound healing between the clean and sterile groups. Mean cost was significantly less for the clean group than for the sterile group. CONCLUSION: These pilot study data show no difference in rate of wound healing with clean versus sterile technique, and clean technique is less expensive. These findings need to be confirmed with a larger sample; type II error cannot be ruled out. PMID- 9204847 TI - Effects of a pressure-reduction mattress and staff education on the incidence of nosocomial pressure ulcers. AB - PURPOSE: The purpose of this study was to determine whether replacing standard hospital mattresses with pressure-reduction mattresses and educating the patient care team on Agency for Health Care Policy and Research prevention guidelines would decrease the incidence of nosocomial pressure ulcers. DESIGN: Retrospective chart review before and after implementation of replacement of standard hospital mattresses with pressure-reduction mattresses and before and after patient care team education was completed. SETTING AND SUBJECTS: A 6-month clinical study with 141 subjects was conducted on a skilled-nursing unit. METHODS: The 3-month preintervention sample group of 141 subjects received routine nursing care on a standard hospital mattress. After the introduction of pressure-reduction mattresses and an education program, a 3-month postintervention sample group of 141 subjects was studied. MAIN OUTCOME MEASURES: Incidence of nosocomial pressure ulcers during a 3-month period. RESULTS: Among the preintervention group, 21 of 141 subjects (15%) acquired nosocomial pressure ulcers, versus 16 of 141 subjects (11%) in the postintervention group. This improvement was not statistically significant (chi 2 = 0.78, df = 1, p = 0.38). The incidence of ulcers staged II or higher dropped from 11 patients in the preintervention group to six in the postintervention group. a 45% reduction that was not statistically significant (chi 2 = 1.56, df = 1, p = 0.21). CONCLUSION: Replacement of standard hospital mattresses and education of staff according to recommendations from the Agency for Health Care Policy and Research guideline for pressure ulcer prediction and prevention did not significantly change the incidence of pressure ulcers during a 3-month period in our skilled-nursing unit. PMID- 9204848 TI - Managing skin care with the CareMap system. AB - In the last three decades, WOC nursing has evolved into on increasingly complex practice. The CareMap is a tool designed to assist in management of patient care. This article defines the CareMap system and describes the development, implementation, and pilot evaluation of a skin care pathway created with the CareMap at Western Pennsylvania Hospital. PMID- 9204849 TI - Biofeedback reeducation in gracilis muscle transposition after rectal trauma: a case presentation. AB - A 20-year-old male patient had a significant rectal trauma and underwent colostomy and subsequent creation of a neosphincter through gracilis muscle transposition. The patient was referred for biofeedback training to learn to use the transplanted gracilis muscle as a substitute for the external anal sphincter, Through a period of 10 weeks, the patient became able to identify this transplanted muscle and to increase the amplitude and duration of contraction as measured by surface electromyography. Further, resting tone of the neosphincter was increased. The patient gained continence with respect to liquid, gaseous, and solid stool within 26 weeks of the first biofeedback muscle training session. PMID- 9204851 TI - Patient with occipital pressure ulcer requiring continued cervical collar use. PMID- 9204850 TI - Clinical experience with a balloon-tipped urethral insert for stress urinary incontinence. AB - A balloon-tipped urethral insert for control of urinary incontinence in women has undergone clinical trials and has been accepted for clinical use by the U.S. Food and Drug Administration. On the basis of results of a multicenter clinical trial, it was concluded that the device provides a safe and effective option for management of genuine stress incontinence and mild mixed incontinence in women. This article reviews appropriate patient selection, education, and training to optimize patient acceptance and efficacy of this urinary control insert. PMID- 9204852 TI - WOCN and the wound healing society: collaboration for wound care. PMID- 9204853 TI - Bridging the gap from bench to bedside in wound care. PMID- 9204854 TI - The development and implementation of an integrated multidisciplinary clinical pathway. AB - Clinical pathways, linear time-related representations of patient care processes, are widely encouraged as a mechanism to outline efficient, cost-effective, multidisciplinary care. The translation of pathways from concept to reality is, however, predictably difficult. All caregivers are dedicated to a common goal, but organizational, personal, and professional perspectives are barriers to development of a common tool. Moreover, the building process requires the discovery, articulation, and communication of previously tacit patient care processes. Although no prescription can work for all pathway development, some strategies can help ensure the best possible chance of pathway success. Participants must recognize that between-patient variability can be expected to decrease with pathway implementation, and educational processes must support that aim. "Stakeholder groups" must be identified and their investment must be assessed, with careful attention paid to acquiring the unconditional support of institutional leadership. Planning of precise building, implementation, and piloting processes, with provision for facilitation of building and implementation processes, is critical. Those charged with pathway development must commit to the establishment and explication of clear goals (economic and quality outcomes) and to careful integration of the pathway with planned and existing continuous quality improvement processes. These strategies are illustrated with actual experiences in implementing cystectomy and pressure ulcer pathways in one academic medical center. PMID- 9204855 TI - Development and implementation of a clinical pathway for radical cystectomy and urinary system reconstruction. AB - A multidisciplinary pathway and patient guide for radical cystectomies is described. Various forms of urinary diversion may be employed after cystectomy for bladder cancer. A clinical pathway for the management of patients undergoing radical cystectomy and urinary diversion or neobladder construction has proved beneficial to patient care, to the nursing and medical staffs, and to the institution. PMID- 9204856 TI - Oral care and its role in WOC nursing. AB - The impact of nutritional status and the ability of the patient to fight infection directly are related to the practice of the WOC nurse, who is well versed in the care of the body's integument with respect to patients with ostomies, wounds, tubes, and incontinence. The status of the patient's oral cavity and its relationship with nutrition and wound healing have not; however been adequately addressed by WOC nurses. Involvement of the WOC nurse in preventive and therapeutic oral care can ensure a healthier gastrointestinal tract, improved nutritional status, and improved outcomes in caring for the person with a wound ostomy, or incontinence. This article reviews the literature related to oral care and discusses its relevance to WOC nursing practice. PMID- 9204857 TI - Skin integrity in patients undergoing prolonged operations. AB - OBJECTIVE: The purpose of this study was to identify risk factors contributing to pressure ulcer development in patients undergoing scheduled, prolonged operative procedures. DESIGN: A descriptive study was conducted. SETTING AND SUBJECTS: A large university teaching facility provided the setting. Thirty-three subjects who underwent operative procedures lasting longer than 10 hours, as determined from the daily operating room schedule through a 6-month period, were included in the study. INSTRUMENTS: Braden Scale for Predicting Pressure Sore Risk was used before the operation. Visual skin inspection, preoperative interventions, and demographic information were documented with a data-collection tool. Postoperative skin breakdown and its severity were assessed as stage I through IV according to the Pressure Ulcer Classification System recommended by the National Pressure Ulcer Advisory Panel. METHODS: Visual preoperative skin assessment was performed and the Braden Scale was completed in the operating room holding area. Demographic information was collected from patient interviews and the medical record. Patient positioning and the placement of all positioning and thermal devices were observed and recorded in the operating room. Within 48 hours after the surgical procedure, the patients' skin was visually inspected. Pressure ulcers were noted, staged, and recorded. MAIN OUTCOME MEASURES: The chi 2 analyses compared those who did and those who did not acquire pressure ulcers for differences in gender, type of operation, position used in the operating room, and types of positioning devices. Student's t tests compared those who did and did not acquire pressure ulcers for differences in age, Braden Scale score, number of positioning devices, and length of operation. RESULTS: Of the 33 patients studied, 15 (45%) were found to acquire stage I or II pressure ulcers within 48 hours after their procedure. Of the 15 patients who acquired pressure ulcers, 75% were placed on a warming blanket during the procedure. This was the only significant finding among the risk factors investigated in the comparison of those who did and did not acquire pressure ulcers (chi 2 = 4.3, p < 0.05). CONCLUSIONS: Removal of the warming blanket from routine intraoperative use with patients undergoing prolonged operations is indicated. Continued follow-up of this patient population will help to determine whether avoidance of warming blankets is sufficient to lower the incidence of pressure ulcer formation. PMID- 9204858 TI - The pursuit of colostomy continence. AB - The lifelong management required by patients with permanent colostomies leads to dissatisfaction with quality of life for many. Through the years, multiple techniques have been attempted to improve the quality of life by pursuing colostomy continence. Such endeavors include surgical interventions, nonsurgical devices and management, and behavior modification techniques. Efforts in research and development continue, and the desire to achieve continence of the stoma remain common cause among persons with on ostomy and those involved in their care. PMID- 9204859 TI - Pelvic muscle rehabilitation: where do we go from here? AB - Pelvic muscle rehabilitation has been used in the treatment of stress and urge urinary incontinence for many years. This article reviews the anatomy of the female pelvic floor and its role in the maintenance of continence. The purpose and goals of pelvic muscle rehabilitation are reviewed, and implications for future applications and research are discussed. PMID- 9204860 TI - A historical review of selected nursing and medical literature on urinary incontinence between 1850 and 1976. AB - The nursing and medical literature on urinary incontinence from 1850 to 1976 was reviewed to provide a historic perspective on care patterns before the current surge in interest in this common condition. Relevant nursing and medical journals and a number of textbooks from both fields were systematically examined to document the evolution of treatments and practices regarding urinary incontinence. Throughout the article, findings are examined in light of the broader historical context to reveal how and why practices were favored or disfavored at given times during the years under investigation. As expected, attitudes, values, and practices in the field reflected the state of knowledge and beliefs commonly held by nurses, physicians, and the general population. PMID- 9204861 TI - Patient with jejunostomy, caput medusae, and bleeding peristomal varices. PMID- 9204862 TI - My two cents on the issue of clean versus sterile wound care. PMID- 9204863 TI - Subunit combinations defined for K+ channel Kv1 subtypes in synaptic membranes from bovine brain. AB - Voltage-dependent Shaker-related (Kv1) K+ channels are composed of transmembrane alpha subunits and peripheral Kv beta proteins that exist as octomers with (alpha)4(beta)4 stoichiometry. Although several alpha (designated Kv1.X) and three Kv beta subunits are known to be expressed in brain, their oligomeric combinations in neurons have yet to be deciphered. Herein, the subunits comprising a number of neuronal K+ channels from bovine brain cortex were deduced by immunoprecipitation and Western blotting, using antibodies specific for Kv1.X and Kv beta subtypes. Only a subset of the theoretically possible oligomers was detected, showing that the synthesis and/or assembly of these multisubunit K+ proteins is controlled to yield a limited variety of K+ channels. Except for a small population of Kv1.4 containing K+ channels, all the recognizable species contained Kv1.2 and beta2 subunits. Furthermore, several subpopulations were identified including a fully defined complex of Kv1.2/1.3/1.4/1.6 and Kv beta2, plus oligomers containing three or two assigned alpha subunits. Kv1.2 was also shown to occur in the absence of these other subunits as a putative homo oligomer. Thus, for the first time, the complete subunit combination of an authentic K+ channel has been elucidated; also, the strategy employed to establish this can now be applied to closely related members of other K+ channel families. PMID- 9204864 TI - Mutation of tyrosine 34 to phenylalanine eliminates the active site pK of reduced iron-containing superoxide dismutase. AB - We have compared the magnetic resonance properties and pH dependence of wild-type and mutant Fe-containing superoxide dismutase (Fe-SOD) in which the conserved active site tyrosine (Tyr 34) is replaced by phenylalanine. The EPR spectrum of the oxidized state and the NMR spectrum of the paramagnetically shifted resonances of the reduced state indicate that in both states the active site is relatively unperturbed by the mutation. Similarly, the mutant Fe-SOD retains approximately 41% of wild-type catalytic activity on a per Fe basis. However, replacement of Tyr 34 by Phe abolishes both NMR spectroscopic signatures of the active site pK of 8.5 of (reduced) Fe2+-SOD. Neither accessibility to base catalyzed exchange nor the chemical shifts of active site residues are affected by pH in the range of 6.5-10.5 in Y34F Fe2+-SOD. Thus, the active site pK of 8.5 of Fe2+-SOD most likely corresponds to deprotonation of Tyr 34. The widespread chemical shift changes associated with the pK could reflect Tyr 34's participation in the active site hydrogen bonding network and the network's propagation of the effects of deprotonating Tyr 34 to the Fe2+. Deprotonation of Tyr 34 can also explain the dramatic decrease in active site accessibility to base-catalyzed exchange as the result of electrostatic repulsion between the exchange catalyst OH- and the (Tyr 34)- ion formed at high pH. Similar electrostatic repulsion between (Tyr 34)- and the substrate O2.- is also consistent with the observed increase in KM above pH 9. PMID- 9204865 TI - Mutational analysis of substrate recognition by protein phosphatase 1. AB - The role of residues that are involved in substrate recognition by rabbit muscle protein phosphatase 1alpha (PP1) was investigated by site-directed mutagenesis and kinetic analyses using phosphorylase a, RII peptide, Kemptide, and p nitrophenyl phosphate as substrates. The atomic structure of PP1 has shown the active site to be at the confluence of three shallow grooves, a C-terminal groove, an acidic groove, and a hydrophobic groove. Mutations of residues D208, D210, D212, E218, D220, E252, D253, E256, E275, and D277 in the acidic groove, of R221, W206, and Y134, which have been suggested to be involved in substrate binding, and of residues C127, I130, and D197 in the hydrophobic groove were examined. Our results show that mutations in the acidic groove lead to modest changes in substrate binding, consistent with a role of the acidic residues in forming a negatively charged surface well for binding of peptides with basic N termini. Severe effects on Vmax were observed for mutants of R221, D208, and W206. These results are consistent with the proposal that the R221 plays an important role as a phosphate oxygen ligand that positions the substrate for catalysis. The kinetic behavior of mutants at W206 and D208 can be explained by the observation that, together with R221, these residues form the microenvironment which dictates the orientation of the imidazole ring of H248, one of the metal binding ligands, as well as contributing to the orientation of R221 itself. PMID- 9204866 TI - Pertussis toxin: transition state analysis for ADP-ribosylation of G-protein peptide alphai3C20. AB - Pertussis toxin from Bordetella pertussis is one of the ADP-ribosylating toxins which are the cytotoxic agents of several infectious diseases. Transition state analogues of these enzymes are expected to be potent inhibitors and may be useful in therapy. Pertussis toxin catalyzes the ADP-ribosylation of a cysteine in the synthetic peptide alphai3C20, corresponding to the C-terminal 20 amino acids of the alpha-subunits of the G-protein Gi3. A family of kinetic isotope effects was determined for the ADP-ribosylation reaction, using 3H-, 14C- and 15N-labeled NAD+ as substrates. Primary kinetic isotope effects were 1.050 +/- 0.006 for [1'N 14C] and 1.021 +/- 0.002 for [1N-15N], the double primary effect of [1'N-14C,1N 15N] was 1.064 +/- 0.002. Secondary kinetic isotope effects were 1.208 +/- 0.014 for [1'N-3H], 1.104 +/- 0.010 for [2'N-3H], 0.989 +/- 0.001 for [4'N-3H], and 1.014 +/- 0.002 for [5'N-3H]. Isotope trapping experiments yielded a commitment factor of 0.01, demonstrating that the observed isotope effects are near intrinsic. Solvent D2O kinetic isotope effects are inverse, consistent with deprotonation of the attacking Cys prior to transition state formation. The transition state structure was determined by a normal mode bond vibrational analysis. The transition state is characterized by a nicotinamide leaving group bond order of 0.14, corresponding to a bond length of 2.06 A. The incoming thiolate nucleophile has a bond order of 0.11, corresponding to 2.47 A. The ribose ring has strong oxocarbenium ion character. Pertussis toxin also catalyzes the slow hydrolysis of NAD+ in the absence of peptides. Comparison of the transition states for NAD+ hydrolysis and for ADP-ribosylation of peptide alphai3C20 indicates that the sulfur nucleophile from the peptide Cys participates more actively as a nucleophile in the reaction than does water in the hydrolytic reaction. Participation of the thiolate anion at the transition state provides partial neutralization of the cationic charge which normally develops at the transition states of N-ribohydrolases and transferases. Thus, the presence of the peptide provides increased SN2 character in a loose transition state which retains oxocarbenium character in the ribose. PMID- 9204867 TI - The QM protein associates with ribosomes in the rough endoplasmic reticulum. AB - QM is a human cDNA originally isolated as a transcript elevated in a nontumorigenic Wilms' tumor microcell hybrid, relative to the tumorigenic parental cell line. Homologs of this gene have been identified from a large number of diverse eukaryotic species which demonstrate a high degree of conservation. The functional importance implied by this strong conservation is supported by the observation that the disruption of the yeast homolog is lethal. In spite of its apparent importance, the function of the encoded protein remains elusive. Indirect immunofluorescent cell staining of cultured human, G401 cells with an antibody to the QM protein shows a punctate staining pattern in the cytoplasm with much of the signal in a perinuclear pattern. Subcellular fractionation demonstrated an association of QM protein with the rough endoplasmic reticulum. It was possible to disrupt this association by washing microsomal membranes with 1M NaCl, suggesting a peripheral association. Proteolytic latency studies showed the protein to be exposed on the cytoplasmic face of the membrane. In situ cross-linking followed by diagonal SDS gel analysis indicates that QM exists as a member of a large protein complex. In agreement with this, QM was found to copurify with the ribosome complex. Incubation with 1 M NaCl was found to disrupt this association while having no effect on the association of core ribosomal proteins. PMID- 9204868 TI - Mechanism of ribose 2'-group discrimination by an RNA polymerase. AB - The mechanism by which T7 RNA polymerase (RNAP) discriminates between rNTP and dNTP substrates has been characterized. During transcript elongation T7 RNAP uses rNTPs 70-80-fold more efficiently than dNTPs. Discrimination of the hydrogen bonding character of the ribose 2'-substituent contributes a largely Km-mediated factor of approximately 20 to this preference for rNTPs. Discrimination of 2' substituent H-bonding character appears to be made through a hydrogen bond to the hydroxyl group of tyrosine 639. This hydrogen bond makes little net contribution to either rNTP ground or transition state binding energy apparently because it is balanced by the energy of desolvation of the tyrosine hydroxyl. This mechanism may reflect a strategy to facilitate translocation by minimizing contributions from polymerase-NMP moiety interactions to NTP binding energy so as to minimize the affinity of the NTP binding site for the 3'-NMP of the product nucleic acid. PMID- 9204869 TI - Unique push-pull mechanism of dealkylation in soman-inhibited cholinesterases. AB - The pH-dependence and solvent isotope effects of dealkylation in diastereomeric adducts of Electric eel (Ee) and fetal bovine serum (FBS) acetylcholinesterase (AChE) inactivated with P(-)C(+) and P(-)C(-) 2-(3,3-dimethylbutyl) methylphosphonofluoridate (soman) were studied at 4.0 +/- 0.2 degrees C. The rate constant versus pH profiles were fit to a bell-shaped curve for all adducts. Best fit parameters are pK1 4.4-4.6 and pK2 6.3-6.5 for Ee AChE and pK1 4.8-5. 0 and pK2 5.8 for FBS AChE. The pKs are consistent with catalytic participation of the Glu199 anion and HisH+440. Maximal rate constants (kmax) are 13-16 x 10(-3) s-1 for Ee AChE and 8 x 10(-3) s-1 for FBS AChE. The solvent isotope effects at the pH maxima are 1.1-1.3, indicating unlikely proton transfer at the enzymic transition states for the dealkylation reaction. Slopes of log rate constant versus pH plots are near 1 at 25.0 +/- 0.2 degrees C between pH 7.0 and 10.0. In stark contrast, the corresponding adducts of trypsin are very stable even at 37.0 +/- 0.2 degrees C. The rate constants for diastereomers of soman-inhibited trypsin at 37.0 +/- 0.2 degrees C are pH independent and approximately 10(4) times smaller than kmax for analogous adducts with AChE. Dealkylation in soman inhibited AChEs is estimated to occur at >10(10) times faster than a plausible nonenzymic reaction. Up to 40% of the catalytic acceleration can be attributed to an electrostatic push, and an electrostatic pull provides much of the balance. The results of this work together with results of a product analysis by Michel et al. (1969) can be explained by an initial and rate-determining methyl migration from Cbeta to Calpha. This is driven by the high electron density of residues (Glu199 and Trp84) at a crowded active site and may be concerted with C-O bond breaking. The positive charge at the rate-determining transition state is distributed between Cbeta and His440. A tertiary carbocation may have a fleeting existence before it is trapped by water or neighboring electrons which is likely to be promoted by Glu199 as the proton acceptor. PMID- 9204870 TI - Mutagenesis of the uncoupling protein of brown adipose tissue. Neutralization Of E190 largely abolishes pH control of nucleotide binding. AB - For expression in Saccharomyces cerevisiae the cDNA of the uncoupling protein (UCP) of brown adipose tissue from hamster has been isolated and used to transform yeast cells. Optimized expression conditions yielded 2% of mitochondrial protein as UCP. UCP was isolated, avoiding copurification of ADP/ATP carrier and porin. Intrahelical E190, previously suggested to be the pH sensor for nucleotide binding, was neutralized to glutamine by mutagenesis. In binding titrations with [14C]guanosine 5'-triphosphate (GTP) and with fluorescent dansyl-GTP, near equal binding capacity for GTP was measured in wild-type (wt) and E190Q. The KD for GTP binding to UCP from yeast has the same strong pH dependence as the original UCP from hamster. With both [14C]GTP and dansyl-GTP, the KD in wt increased 16-19-fold from pH 6.0 to 7.5, while in E190Q this increase was only 2.5-2.9-fold. As a result, at pH 7.5, both [14C]GTP and dansyl GTP bind 6-fold tighter to E190Q than to wt. The binding rate of GTP decreased 10 fold from pH 6.0 to 7.5 in wt and only 4-fold in E190Q. Woodward reagent K (WRK) known to interact specifically with E190 [Winkler, E., Wachter, E., and Klingenberg, M. (1997) Biochemistry 36, 148-155] abolished [14C]GTP and dansyl GTP binding to wt UCP, whereas binding to E190Q was fully resistant to WRK. H+ and Cl- transport activity in reconstituted vesicles were the same with wt and E190Q. At pH 7.5, 5 microM GTP is unable to inhibit H+ and Cl- transport in wt but inhibits in E190Q to maximum level. The different sensitivity toward GTP versus GDP found in wt is absent in E190Q. Thus, the mutation E190Q results in the predicted gain of function in binding and proves the role of the intrahelical E190 as a pH sensor for nucleotide binding but excludes a role in transport. PMID- 9204871 TI - Membrane location of spin-labeled M13 major coat protein mutants determined by paramagnetic relaxation agents. AB - Mutants of the M13 bacteriophage major coat protein containing single cysteine replacements (A25C, V31C, T36C, G38C, T46C, and A49C) in the hydrophobic and C terminal domains were purified from viable phage. These were used for site directed spin-labeling to determine the location and assembly of the major coat protein incorporated in bilayer membranes of dioleoylphosphatidylcholine. The membrane location of the spin-labeled cysteine residues was studied with molecular oxygen and Ni2+ ions as paramagnetic relaxation agents preferentially confined to the hydrophobic and aqueous regions, respectively, by using progressive-saturation electron spin resonance (ESR) spectroscopy. The section of the protein around Thr36 is situated at the center of the membrane. Residue Thr46 is placed at the membrane surface in the phospholipid head group region with a short C-terminal section, including Ala49, extending into the aqueous phase. Residue Ala25 is then positioned consistently in the head group region of the apposing lipid monolayer leaflet. These positional assignments are consistent with the observed mobilities of the spin-labeled groups. The outer hyperfine splittings in the ESR spectra decrease from the N-terminal to the C-terminal of the hydrophobic section (residues 25-46), and then drop abruptly in the aqueous phase (residue 49). Additionally, the strong immobilization and low oxygen accessibility of residue 25 are attributed to steric restriction at the hinge region between the transmembrane and N-terminal amphipathic helices. Sequence specific modulations of the ESR parameters are also observed. Relatively low oxygen accessibilities in the hydrophobic region suggest intermolecular associations of the transmembrane helices, in agreement with saturation transfer ESR studies of the overall protein mobility. Relaxation enhancements additionally reveal a Ni2+ binding site in the N-terminal domain that is consistent with a surface orientation of the amphipathic helix. PMID- 9204872 TI - The modified wobble base inosine in yeast tRNAIle is a positive determinant for aminoacylation by isoleucyl-tRNA synthetase. AB - Earlier work by two independent groups has established the fact that anticodons GAU and LAU of Escherichia coli tRNAIle isoacceptors play a critical role in the tRNA identity. Yeast possesses two isoleucine transfer RNAs, a major one with anticodon IAU and a minor one with anticodon PsiAPsi which are derived from the post-transcriptional modification of AAU and UAU gene sequences, respectively. We present direct evidence which reveals that inosine is a positive determinant for yeast isoleucyl-tRNA synthetase. We also show that yeast tRNAMet with guanosine at the wobble position becomes aminoacylated with isoleucine while methionine acceptance is lost. As inosine and guanosine share the 6-keto and the N-1 hydrogen groups, this suggests that these hydrogen donor and acceptor groups are determinants for isoleucine specificity. The role of the minor tRNAIle anticodon pseudouridines in tRNA isoleucylation could not be tested directly but was deduced from a 40-fold decrease in the activity of the unmodified transcript. The presence of the NHCO structure in guanosine, inosine, pseudouridine, and lysidine suggests a unifying model of wobble base recognition by the yeast and E. coli isoleucyl-tRNA synthetase. In contrast to lysidine which switches the identity of the tRNA from methionine to isoleucine [Muramatsu, T., Nishikawa, K., Nemoto, F., Kuchino, Y., Nishimura, S., Miyazawa, T., & Yokoyama, S. (1988) Nature 336, 179 181], pseudouridine-34 does not modify the specificity of the yeast minor tRNAIle since U-34 is a strong negative determinant for yeast MetRS. Therefore, the major role of Psi-34 (in combination with Psi-36 or not) is likely in isoleucine AUA codon specificity and translational fidelity. PMID- 9204873 TI - The scaffold/matrix attachment region binding protein hnRNP-U (SAF-A) is directly bound to chromosomal DNA in vivo: a chemical cross-linking study. AB - The protein heterogeneous nuclear ribonucleoprotein U (hnRNP-U, also known as scaffold attachment factor A, SAF-A) is an abundant component of hnRNP particles and of the nuclear matrix. Previous experiments have demonstrated that, in vitro, hnRNP-U specifically binds to scaffold/matrix attachment (S/MAR) region DNA elements and could thus be involved in higher order chromatin structure. In this paper we report on the use of chemical cross-linking to investigate whether the protein is also bound to DNA in vivo, which is a prerequisite for its presumed function in chromatin loop formation. We have improved published methods for cross-linking proteins to DNA with the aim to minimize unspecific fixation and possible contamination with RNA binding proteins. Our protocol is based on a limited cross-linking of living human cells with formaldehyde, followed by the purification of DNA/protein complexes by two consecutive cesium chloride density gradient centrifugations. Analysis of the protein constituents of these complexes shows a specific subset of cross-linked proteins with the histones as major components. By western blotting, we demonstrate that hnRNP-U is efficiently cross linked to DNA under experimental conditions that yield DNA/protein complexes with a buoyant density equivalent to that of native chromatin. Dimethylsulfate cross linking and limited protease digestion of the complexes was used to establish that hnRNP-U is bound directly to DNA and not via cross-linking to other proteins. This is the first direct demonstration of the in vivo DNA binding of a S/MAR specific protein and suggests a structural role of hnRNP-U in chromatin organization. PMID- 9204874 TI - The PvuII DNA (cytosine-N4)-methyltransferase comprises two trypsin-defined domains, each of which binds a molecule of S-adenosyl-L-methionine. AB - Earlier studies have shown that PvuII methyltransferase is monomeric and transfers a methyl group from S-adenosyl-l-methionine (AdoMet) to cytosine, generating N4-methylcytosine in duplex 5'-CAGCTG-3' DNA. This study examines the interactions between PvuII methyltransferase and AdoMet. Trypsin preferentially cleaved the protein into two large fragments, with initial cleavages after Arg183 and Lys186. UV-mediated photochemical labeling with [3H-CH3]AdoMet, followed by trypsin digestion, revealed that both large fragments of the protein were labeled. Rapid gel filtration confirmed that each molecule of the intact enzyme bound two molecules of AdoMet (net Kd = 9.3 microM). When PvuII methyltransferase was preincubated with a range of [3H-CH3]AdoMet concentrations, bursts of product formation resulted upon DNA addition. These data indicate that PvuII methyltransferase is catalytically competent with one and with two bound molecules of AdoMet. These results, together with those from earlier studies, suggest possible roles for the second molecule of AdoMet. PMID- 9204876 TI - The 60 kDa insulin receptor substrate functions like an IRS protein (pp60IRS3) in adipose cells. AB - The 60 kDa insulin receptor substrate in rat adipocytes that binds to the PI-3 kinase displays several functional characteristics in common with the IRS proteins; so we propose the name pp60(IRS3) to distinguish it from other tyrosine phosphorylated proteins of similar size. During insulin stimulation, p85 associated with pp60(IRS3) more rapidly than with IRS-1 or IRS-2. In mice lacking IRS-1, p85 associated more strongly with pp60(IRS3) than with IRS-2, suggesting that pp60(IRS3) provides an alternate pathway in these cells. Synthetic peptides containing two phosphorylated YMPM motifs displace pp60(IRS3) and IRS-1 from alphap85 immune complexes, suggesting that pp60(IRS3), like IRS-1, engages both SH2 domains in p85. Moreover, pp60(IRS3) binds to immobilized peptides containing a phosphorylated NPXY motif, suggesting that it contains a PTB domain with similar specificity to that in IRS-1. The cloning of pp60(IRS3) will reveal a new member of the IRS protein family which mediates insulin receptor signals in a narrow range of tissues. PMID- 9204875 TI - Effects of divalent metal ions on individual steps of the Tetrahymena ribozyme reaction. AB - The Tetrahymena thermophila L-21 ScaI ribozyme utilizes Mg2+ to catalyze a site specific endonuclease reaction analogous to the first step of self-splicing. To better understand the contribution of Mg2+ to ribozyme activity, the Mg2+ concentration dependence of individual rate constants was examined at concentrations greater than those required for ribozyme folding (>2 mM; at 50 degrees C and pH 6.7). Analysis of metal ion inhibition of the chemical step of the reaction indicated that two Ca2+ ions compete with two Mg2+ ions involved in active site chemistry. These Mg2+ ions are bound tightly to the E.S complex (Kd < 2 mM). The rate constant for association of the oligoribonucleotide substrate (S) increased 12-fold from 2 to 100 mM Mg2+ and exhibited saturation behavior, consistent with a single Mg2+ ion involved in S association that binds to the free ribozyme with a Kd for Mg2+ of 15 mM. The preference for the divalent metal ion (Mg2+ congruent with Ca2+ > Ba2+ >> Sr2+) suggested that enhancing the rate constant of S association is not simply a function of ionic strength, but is due to a distinct metal ion binding site. Even though Ca2+ does not support reaction, the RNA substrate S was able to bind in the presence of Ca2+. Upon addition of Mg2+, S was cleaved without first dissociating. A model is proposed in which the inactive Ca2+ form of E.S is structurally equivalent to the open complex along the reaction pathway, which has the RNA substrate bound but not docked into the active site. Weaker binding of S in Ca2+ was shown to result from an increase in the rate constant of S dissociation, leading to the proposal that a tight Mg2+ binding site or sites in the E.S complex contribute to the strong binding of S. In summary, the data provide evidence for four functions for bound Mg2+ ions in the catalytic cycle: one increases the rate of RNA substrate binding, one or more decrease the rate of dissociation of S, and two are involved in the chemical step. PMID- 9204877 TI - Adipocyte fatty acid-binding protein: interaction with phospholipid membranes and thermal stability studied by FTIR spectroscopy. AB - Fatty acid-binding proteins (FABPs) found in many tissues constitute a family of low molecular weight proteins that are suggested to function as intracellular transporters of fatty acids. Studies of the transfer kinetics of fluorescent anthroyloxy-labeled long-chain fatty acids from FABP to model membranes led to the suggestion that the FABPs, typically considered to be cytosolic proteins, could nevertheless interact directly with membranes [Wootan, M. G., et al. (1993) Biochemistry 32, 8622-8627]. In the current study, the interaction of the adipocyte FABP (A-FABP) with vesicles of various phospholipids has been directly measured and confirmed with FTIR spectroscopy. The strength of this interaction was inferred from the lowering of the gel-liquid-crystal phase transition temperature as monitored from temperature-induced variations in the acyl chain CH2 stretching frequencies and from the intensities of the components of the CH2 wagging progressions. A-FABP interacts more strongly with anionic phospholipids (phosphatidylserine and cardiolipin) than with zwitterionic phosphatidylcholine. Unsaturation in the acyl chains leads to a greater reduction in Tm (stronger lipid-protein interaction). In contrast, neutralization of A-FABP surface charges by acetylation considerably weakens the interaction. Comparison of the shifts in lipid melting temperatures with those induced by other proteins suggests that A FABP behaves like a typical peripheral membrane protein. The degree of membrane interaction correlates directly with the rate of fatty acid transfer, suggesting that contact between A-FABP and membranes is functionally related to its fatty acid transport properties. As expected, the protein exhibits a predominantly beta sheet structure. It was found to aggregate with increasing temperature. With the exception of minor differences between the pure and lipid-associated A-FABP in the 1640-1660 cm-1 region, both the protein structure and thermal stability appeared essentially unchanged upon interaction with the lipid. PMID- 9204878 TI - Targeting of the catalytic subunit of protein phosphatase-1 to the glycolytic enzyme phosphofructokinase. AB - In this study, we demonstrate that the catalytic subunit of rabbit muscle protein phosphatase-1 (PP1) binds to muscle phosphofructokinase (6-phosphofructo-1 kinase, PFK). A protein of 85 kDa was isolated from rat muscle by affinity chromatography on PP1-Sepharose and was identified as phosphofructokinase by partial amino acid sequence analysis. This novel finding of a protein-protein interaction between PP1 and PFK was confirmed by reciprocal experiments in which the binding of PP1 to PFK-agarose was demonstrated. Elution of PP1 from PFK agarose was maximal at ca. 0.4 M NaCl. The specificity of binding was demonstrated by isolation of PP1 from a partially purified rabbit muscle PP1 preparation. All four known isoforms of PP1 (PP1alpha, PP1gamma1, PP1gamma2, and PP1delta) were shown to bind to PFK-agarose. The activity of PP1 was only partially inhibited by PFK. The preformed complex between PP1 and PFK did not bind to inhibitor-2-Sepharose. The stoichiometry of binding of PP1 to the PFK monomer was found to be 1:1 in the isolated PP1.PFK complex. An interaction between PP1 and PFK in muscle extracts was demonstrated by their coimmunoprecipitation. Our findings raise the interesting possibility that PP1 may be targeted to PFK, and may be physiologically relevant in the context that PFK and other glycolytic enzymes have been shown to be micro-compartmentalized by binding to F-actin. This in turn points to a role for PP1 in control of glycolytic flux by protein phosphorylation-dephosphorylation mechanisms. PMID- 9204879 TI - Two mechanisms for the recapture of extracellular GM2 activator protein: evidence for a major secretory form of the protein. AB - The GM2 activator protein is a small monomeric protein containing a single site for Asn-linked glycosylation. Its only proven in vivo function is to act as a substrate specific cofactor for the hydrolysis of GM2 ganglioside by lysosomal beta-hexosaminidase A. However, we and others have shown it can act as a general glycolipid transporter at neutral pH in vitro. Any other possible in vivo functions would require that some of the newly synthesized activator molecules not be targeted to the lysosome. The lysosomal targeting mechanism for the activator has not been conclusively identified. While earlier reports suggested that it is likely through the mannose-6-phosphate receptor, another more recent report demonstrated that deficient human cells could recapture nonglycosylated, bacterially produced activator, suggesting its use of an alternate targeting pathway. Here, we demonstrate that the mannose-6-phosphate pathway is likely the major intracellular, biosynthetic route to the lysosome, as well as a high affinity recapture pathway for the endocytosis of activator protein from extracellular fluids. Additionally, we show that there exists a second lower affinity recapture pathway that requires its native protein structure, is carbohydrate independent, and likely does not involve its ability to bind glycosphingolipids in the plasma membrane. Finally, we document that the pool of newly synthesized precursor activator protein contains a majority of molecules with a complex-type oligosaccharide, which cannot contain a functional mannose-6 phosphate targeting signal. These molecules makeup the secreted forms of the protein in normal human fibroblasts. PMID- 9204880 TI - Pre-steady-state kinetic analysis of the trichodiene synthase reaction pathway. AB - The pre-steady-state kinetics of the trichodiene synthase reaction were investigated by rapid chemical quench methods. The single-turnover rate was found to be 3.5-3.8 s-1, a rate 40 times faster than the steady-state catalytic rate (kcat = 0.09 s-1) for trichodiene synthase-catalyzed conversion of farnesyl diphosphate (FPP) to trichodiene at 15 degrees C. In a multiturnover experiment, a burst phase (kb = 4.2 s-1) corresponding to the accumulation of trichodiene on the surface of the enzyme was followed by a slower, steady-state release of products (klin = 0.086 s-1) which corresponds to kcat. These results strongly suggest that the release of trichodiene from the enzyme active site is the rate limiting step in the overall reaction, while the consumption of FPP is the step which limits chemical catalysis at the active site. Single-turnover experiments with trichodiene synthase mutant D101E, for which the steady-state rate constant kcat is 1/3 of that of wild type, revealed that the mutation actually depresses the rate of FPP consumption by a factor of 100. The deuterium isotope effect on the consumption of [1-2H,1,2-14C]FPP was found to be 1.11 +/- 0.06. Single turnover reactions of [1,2-14C]FPP catalyzed by trichodiene synthase were carried out at 4, 15, or 30 degrees C in an effort to provide direct observation of the proposed intermediate nerolidyl diphosphate (NPP). However, no NPP was detected, indicating that the conversion of NPP must be too fast to be observed within the detection limits of the assay. Taken together, these observations suggest that the isomerization of FPP to NPP is the step which limits the rate of chemical catalysis in the trichodiene synthase reaction pathway. PMID- 9204881 TI - Pre-steady-state study of recombinant sesquiterpene cyclases. AB - An Escherichia coli expression system was used to generate hexahistidyl-tagged plant sesquiterpene cyclases, which were readily purified by a single affinity chromatographic step. Genes for Hyoscyamus muticus vetispiradiene synthase (HVS), a chimeric 5-epi-aristolochene synthase (CH3), and a chimeric sesquiterpene cyclase possessing multifunctional epi-aristolochene and vetispiradiene activity (CH4) were expressed in bacterial cells, which resulted in the sesquiterpene cyclases accumulating to 50% of the total protein and 35% of the soluble protein. From initial velocity experiments, the Michaelis constant for HVS was 3.5 microM, while CH3 and CH4 exhibited smaller values of 0.7 and 0.4 microM, respectively. Steady-state catalytic constants were from 0.02 to 0.04 s-1. A combination of pre steady-state rapid quench experiments, isotope trapping experiments, and experiments to measure the burst rate constant as a function of substrate concentration revealed that turnover in all three cyclases is limited by a step after the initial chemical step involving rupture of the carbon-oxygen bond in farnesyl diphosphate (FPP). Rate constants for the limiting step were 10-70-fold smaller than for the initial chemical step. Dissociation constants for the enzyme substrate complex (20-70 microM) were determined from the pre-steady-state experiments and were significantly larger than the observed Michaelis constants. A mechanism that involves an initial, rapid equilibration of enzyme with substrate to form an enzyme-substrate complex, followed by a slower conversion of FPP to an enzyme-bound hydrocarbon and a subsequent rate-limiting step, is proposed for the three enzymes. Interestingly, the multifunctional chimeric enzyme CH4 exhibited both a tighter binding of FPP and a faster conversion of FPP to products than either of its wild-type parents. PMID- 9204882 TI - Plant polyketide synthases leading to stilbenoids have a domain catalyzing malonyl-CoA:CO2 exchange, malonyl-CoA decarboxylation, and covalent enzyme modification and a site for chain lengthening. AB - Stilbene synthases and the related bibenzyl synthases are plant polyketide synthases whose biological functions lie in the formation of antimicrobial phytoalexins. The formation of hydroxystilbenes from one molecule of acyl-CoA and three molecules of malonyl-CoA is catalyzed by a homodimeric 90 kDa protein and includes Claisen condensations and cleavage of a thioester followed by decarboxylation. Combining inhibitor studies, protein modifications, and site directed mutagenesis, we were able to differentiate between the binding sites for malonyl-CoA and the regions responsible for the selection of the primer, p coumaroyl-CoA or m-hydroxyphenylpropionyl-CoA, respectively. Mutations in the C terminal part of the molecule or modification by photolabeling with p azidocinnamoyl-CoA influence the overall reaction, the formation of hydroxystilbenes, but leave partial reactions, such as the malonyl-CoA:CO2 exchange and the malonyl-CoA-dependent modification of the enzyme, unaffected. Data obtained with several kinds of stilbene synthase and mutant forms suggest that the malonyl-CoA-dependent covalent modification takes place at a cysteine residue in the N-terminal part of the enzyme. Mutations in the C-terminal half of the enzyme molecule do not interfere with the malonyl-CoA-dependent reactions. PMID- 9204883 TI - Identification and isolation of a 45-kDa calcium-dependent lactoferrin receptor from rat hepatocytes. AB - Isolated rat hepatocytes bind, internalize, and degrade bovine lactoferrin (Lf) via high-affinity Ca2+-dependent sites [<10(6) sites/cell; McAbee et al., (1993) Biochemistry 32, 13749-13760]. In this study, we identified a 45-kDa Ca2+ dependent Lf binding protein on rat hepatocytes by three independent approaches. First, dithiobis(sulfosuccimidylproprionate) (DTSSP) cross-linked 125I-Lf to a 45 kDa adduct in a Ca2+-dependent manner on intact cells. The 125I-labeled cross linked complexes were absent when either surface-bound 125I-Lf was stripped prior to cross-linking or an excess of unlabeled Lf was included in the DTSSP reaction. Second, 125I-Lf bound to a 45-kDa hepatocyte polypeptide in a Ca2+-dependent fashion following incubation with SDS-PAGE fractioned hepatocyte membrane proteins absorbed on nitrocellulose. Third, when Triton X-100 extracts of hepatocyte membrane ghosts were chromatographed on Lf-agarose, a 45-kDa polypeptide (p45) was eluted by EGTA. Column fractions enriched in p45--but not those depleted of p45--possessed soluble Lf receptor activity as determined by competition binding assay. Monospecific polyclonal anti-p45 IgG detected p45 in crude hepatocyte ghost homogenates and blocked vigorously 125I-Lf binding and endocytosis to intact rat hepatocytes. We conclude, therefore, that p45 constitutes the Ca2+-dependent Lf receptor on isolated rat hepatocytes. PMID- 9204884 TI - Isolated rat hepatocytes bind lactoferrins by the RHL-1 subunit of the asialoglycoprotein receptor in a galactose-independent manner. AB - Isolated rat hepatocytes bind and internalize the iron-binding protein lactoferrin (Lf) by a set of high-affinity, recycling, Ca2+-dependent binding sites. We have purified a 45-kDa membrane protein (p45) from rat hepatocytes that exhibits Ca2+-dependent receptor activity. In this study, we found p45 to be identical to the major subunit (RHL-1) of the rat asialoglycoprotein receptor. Two tryptic fragments of p45 showed 100% identity with RHL-1 internal sequences (Leu121 --> Lys126 and Phe198 --> Lys220), and monospecific antisera against p45 and RHL-1 cross-reacted equally well with each protein. Molar excesses of anti p45 IgG, anti-RHL-1 IgG, asialoorosomucoid, and asialofetuin competitively blocked the binding of 125I-Lf to isolated rat hepatocytes at 4 degrees C. Similarly, either excess anti-p45 or Lf blocked the binding of 125I asialoorosomucoid to cells at 4 degrees C. We did not detect the minor subunits of the rat asialoglycoprotein receptor (RHL-2/3) in p45 preparations from Triton X-100 extracts of hepatocytes and 125I-Lf bound to purified RHL-1 but not to RHL 2/3 immobilized on nitrocellulose. Nonetheless, anti-RHL-2/3 IgG reduced the binding of 125I-Lf to hepatocytes at 4 degrees C. Exoglycosidases were used to remove terminally-exposed N-acetylneuraminyl, alpha- and beta-galactosyl, and N acetylhexosaminyl sugars from human and bovine Lf glycans, and lectin blotting confirmed that glycosidase-treated Lfs lacked detectable terminal galactosyl sugars. Unexpectedly, these deglycosylated Lfs exhibited no loss in their ability to compete with unmodified Lfs for binding to isolated hepatocytes. In addition, molar excess of beta-lactose but not sucrose competitively blocked the binding of 125I-Lf to cells, indicating that Lf bound at or very near the carbohydrate recognition domain of RHL-1. We conclude that RHL-1 is the Ca2+-dependent Lf receptor on hepatocytes and that it binds Lf at its carbohydrate-recognition domain yet in a galactose-independent manner. PMID- 9204885 TI - Specific increase in amyloid beta-protein 42 secretion ratio by calpain inhibition. AB - Cerebral deposition of amyloid beta-protein (Abeta) as senile plaques is a pathological hallmark of Alzheimer's disease (AD). Abeta falls into two major subspecies defined by their C-termini, Abeta40 and Abeta42, ending in Val-40 and Ala-42, respectively. Although Abeta42 accounts for only approximately 10% of secreted Abeta, Abeta42 is the predominant species accumulated in senile plaques in AD brain and appears to be the initially deposited species. Its secretion level has recently been reported to be increased in the plasma or culture media of fibroblasts from patients affected by any of early-onset familial AD (FAD). Thus, inhibition of Abeta42 production would be one of the therapeutic targets for AD. However, there is little information about the cleavage mechanism via which Abeta40 and Abeta42 are generated and its relationship to intracellular protease activity. Here, we examined by well-characterized enzyme immunoassay the effects of calpain and proteasome inhibitors on the levels of Abeta40 and Abeta42 secretion by cultured cells. A calpastatin peptide homologous to the inhibitory domain of calpastatin, an endogenous calpain specific inhibitor, induced a specific increase in secreted Abeta42 relative to the total secreted Abeta level, a characteristic of the cultured cells transfected with FAD-linked mutated genes, while a proteasome specific inhibitor, lactacystin, showed no such effect. These findings suggest that the Abeta42 secretion ratio is modulated by the calpain calpastatin system and may point to the possibility of exploring particular compounds that inhibit Abeta42 secretion through this pathway. PMID- 9204886 TI - Profilin interacts with the Gly-Pro-Pro-Pro-Pro-Pro sequences of vasodilator stimulated phosphoprotein (VASP): implications for actin-based Listeria motility. AB - Intracellular actin-based motility of Listeria monocytogenes requires protein protein interactions involving two different proline-rich sequences: first, the tightly bound bacterial surface protein ActA uses its multiple oligoproline registers [consensus sequence = FE(D)FPPPPTD(E)E(D)] to tether vasodilator stimulated phosphoprotein (VASP) to the bacterial surface; and second, VASP then deploys its own multiple GPPPPP (or GP5) registers to localize the actin regulatory protein profilin to promote actin polymerization. We now report that fluorescence titration showed that GP5GP5GP5 peptide binds to profilin (KD of 84 microM), and the peptide weakly inhibits exchange of actin-bound nucleotide in the absence or presence of profilin. Microinjection of synthetic GPPPPP triplet into Listeria-infected PtK2 cells promptly arrested motility at an intracellular concentration of 10 microM. This inhibition was completely neutralized when equimolar concentrations of profilin and GP5GP5GP5 were simultaneously microinjected. Fluorescence studies with [His-133-Ser]-profilin, a site-directed mutant previously shown to be defective in binding poly-l-proline [Bjorkegren, C., Rozycki, M., Schutt, C. E., Lindberg, U., & Karlsson, R. (1993) FEBS Lett. 333, 123-126], exhibits little or no evidence of saturable GP5GP5GP5 binding. When an equimolar concentration of this [His-133-Ser]-profilin mutant was co injected with GP5GP5GP5, the peptide's inhibitory action remained completely unaffected, indicating that GP5GP5GP5 binding to wild-type profilin represents a key step in actin-based pathogen motility. We also present a model that shows how the focal binding of VASP with its GPPPPP registers can greatly increase the local concentration of profilin and/or profilin-actin-ATP complex at the bacteria/rocket-tail interface. PMID- 9204887 TI - Polyalanine-based peptides as models for self-associated beta-pleated-sheet complexes. AB - The occurrence of beta-sheet motifs in a number of neurodegenerative disorders has brought about the need for the de novo design of soluble model beta-sheet complexes. Such model complexes are expected to further the understanding of the interconversion processes that occur from cellular allowed random coil or alpha helical conformation into insoluble cell-deleterious beta-pleated-sheet motifs. In the present study, polyalanine-based peptides (i.e., derived from Ac-KA14K NH2) were designed that underwent conformational changes from monomeric random coil conformations into soluble, macromolecular beta-pleated-sheet complexes without any covalent modification. The interconversion was found to be length-, environment-, and concentration-dependent and to be driven by hydrophobic interactions between the methyl groups of the alanine side chains. A series of substitution analogs of Ac-KA14K-NH2 was used to study the amino acid acceptability within the hydrophobic core of the complex, as well as at both termini. The formation of amyloid plaques in a number of amyloidogenic peptides could be related to the presence of amino acids within their sequences that were found to have a high propensity to occur in these model beta-sheet complexes. PMID- 9204888 TI - Functional interactions in cytochrome P450BM3: flavin semiquinone intermediates, role of NADP(H), and mechanism of electron transfer by the flavoprotein domain. AB - Cytochrome P450BM3 is a self-sufficient soluble fatty acid hydroxylase from Bacillus megaterium utilizing tightly bound FAD and FMN cofactors to transfer reducing equivalents from NADPH to the heme active site. Active-inactive transitions of cytochrome P450BM3 were exploited to identify catalytic intermediates of the enzyme. Shortly upon reduction by NADPH, a two-electron reduced active P450BM3 is formed with two flavin semiquinones, anionic and neutral, present simultaneously. P450BM3 inactivated by NADPH has a three electron reduced flavoprotein domain. NADPH is unable to reduce P450BM3 rapidly unless the flavoprotein domain is fully oxidized. During steady-state hydroxylation of a poor substrate, tetradecanol, the flavoprotein reduction state does not exceed two, with two flavin semiquinones, anionic and neutral, present. Absorbance and EPR spectroscopic characterization of both anionic and neutral flavin semiquinone is presented. NADPH and NADH were compared as electron donors for P450BM3-catalyzed fatty acid hydroxylation and cytochrome c and heme iron reduction. The Km for NADH of 3-5 mM is about 3000 times higher than the Km of 1 1.5 microM for NADPH. Although NADH can support cytochrome c reduction and fatty acid hydroxylation with the rates as high as 22 and 13 s-1, respectively, these turnover numbers are only about 20% of those observed with NADPH. The results suggest that nucleotide binding plays an important role in catalysis by controlling electron-transfer properties of the flavin cofactors. In W574G and G570D mutant P450BM3 enzymes that are deficient in FMN, NADP+ binding stabilizes fully reduced FAD. P450BM3 catalyzes single-turnover and steady-state laurate hydroxylation with near stoichiometric product formation at NADPH concentrations below that of the enzyme. A mechanism of electron transfer by the flavoprotein domain of P450BM3 is proposed with the reduction state of the flavoprotein domain cycling in a 0-2-1-0 sequence. We also propose that an interaction of bound NADP+ with anionic FAD semiquinone is essential for splitting a pair of electrons that are then transferred in two one-electron transfer steps to the heme catalytic site. PMID- 9204889 TI - Mechanism of helix induction by trifluoroethanol: a framework for extrapolating the helix-forming properties of peptides from trifluoroethanol/water mixtures back to water. AB - To establish a framework for extrapolating the helix-forming properties of peptides from TFE/H2O mixtures (TFE = 2,2, 2-trifluoroethanol) back to water, the thermal unfolding curves have been measured by circular dichroism for four repeating-sequence peptides, with chain lengths from 7 to 22 residues. The unfolding curves were measured between 0 and 50 volume percent TFE and were fitted to the modified Lifson-Roig theory. A single set of helix-coil parameters fits the results for the four peptides at each TFE concentration; only two of the basic helix-coil parameters, , the mean helix propagation parameter of residues in the sequence repeat, and DeltaH, the enthalpy change per residue on unfolding the helix, are allowed to vary with TFE molarity. The success in fitting these curves over a wide range of experimental variables shows that helix formation is basically the same in TFE/H2O mixtures as in water. Moreover, a simple model based on a linear dependence of ln and DeltaH on TFE molarity can be used to extrapolate the results from 25% TFE (approximately 4 M) back to water. The results also give curves of helix formation induced by TFE at constant temperature, and the properties of these helix induction curves explain some of the puzzling results shown by other peptides in the literature. The average helix propensity increases regularly from 0 to 25% TFE but levels off at higher TFE concentrations, which explains why the extent of helix formation levels off in this range. The change in the apparent cooperativity of thermal unfolding curves in concentrated TFE solutions results from the decrease of the enthalpy change for helix unfolding at higher TFE concentrations. The rapid decrease in the plateau values of apparent helix content with increasing temperature results mainly from the strong temperature dependence of the ellipticity of the complete helix. To determine whether the helix-stabilizing effect of TFE arises from strengthening the hydrogen bonds in the helix backbone, the strength of the hydrogen bond in a model compound, salicylic acid, has been measured in TFE/H2O mixtures from the pKa difference between salicylic acid and a similar compound which cannot form the hydrogen bond. The curve of hydrogen bond strength versus increasing TFE concentration matches both in shape and magnitude the increase in average helix propensity in TFE/H2O mixtures. PMID- 9204890 TI - Tetrahydrobiopterin binding to macrophage inducible nitric oxide synthase: heme spin shift and dimer stabilization by the potent pterin antagonist 4-amino tetrahydrobiopterin. AB - The characteristics of tetrahydrobiopterin (H4biopterin) binding to pteridine free recombinant macrophage inducible nitric oxide synthase expressed in Escherichia coli were investigated with a special focus given to effects caused by 2,4-diamino-5,6,7, 8-tetrahydro-6-(l-erythro-1,2-dihydroxypropyl)pteridine (4 amino-H4biopterin), a novel pterin-based inhibitor of nitric oxide synthase. The 4-amino compound completely inhibited enzyme stimulation by 10 microM H4biopterin with a half-maximally active concentration of 7.2 +/- 0.39 microM, whereas H2biopterin and sepiapterin were much less potent. Binding studies using [3H]H4biopterin at 4 degrees C revealed biphasic association of the radioligand according to two first-order reactions with apparent rate constants of 2.2 and 0.05 min-1, each accounting for approximately 50% of total binding. Dissociation of [3H]H4biopterin occurred with rate constants of 0.005 and 0.0028 min-1 in the absence and presence of l-arginine, respectively. Specific binding of 10 nM [3H]H4biopterin was antagonized by unlabeled H4biopterin and its 4-amino analog with half-maximal effects at 84 +/- 6 and 34 +/- 3.2 nM, respectively. Binding of H4biopterin and 4-amino-H4biopterin was accompanied by a partial low spin to high spin conversion of the heme that was completed by l-arginine. Similarly, the active cofactor and the inhibitory 4-amino derivative both induced significant formation of stable protein dimers that survived during SDS electrophoresis, suggesting that the allosteric effects caused by H4biopterin do not explain sufficiently the essential role of the pteridine cofactor in NO biosynthesis. PMID- 9204892 TI - Control of filament formation in Candida albicans by the transcriptional repressor TUP1. AB - The pathogenic yeast Candida albicans regulates its cellular morphology in response to environmental conditions. Ellipsoidal, single cells (blastospores) predominate in rich media, whereas filaments composed of elongated cells that are attached end-to-end form in response to starvation, serum, and other conditions. The TUP1 gene, which encodes a general transcriptional repressor in Saccharomyces cerevisiae, was isolated from C. albicans and disrupted. The resulting tup1 mutant strain of C. albicans grew exclusively as filaments under all conditions tested. TUP1 was epistatic to the transcriptional activator CPH1, previously found to promote filamentous growth. The results suggest a model where TUP1 represses genes responsible for initiating filamentous growth and this repression is lifted under inducing environmental conditions. PMID- 9204893 TI - Modulation of hepatic gene expression by hepatocyte nuclear factor 1. AB - Hepatocyte nuclear factors 1 and 4 (HNF-1 and HNF-4) are liver-enriched transcription factors that function in the regulation of several liver-specific genes. HNF-1 activates genes containing promoters with HNF-1 binding sites. However, this factor negatively regulates its own expression and that of other HNF-4-dependent genes that lack HNF-1 binding sites in their promoter region. This repression is exerted by a direct interaction of HNF-1 with AF2, the main activation domain of HNF-4. The dual functions of gene activation and repression suggest that HNF-1 is a global regulator of the transcriptional network involved in the maintenance of hepatocyte-specific phenotype. PMID- 9204894 TI - Positive effects of combined antiretroviral therapy on CD4+ T cell homeostasis and function in advanced HIV disease. AB - Highly active antiretroviral therapy (HAART) increases CD4(+) cell numbers, but its ability to correct the human immunodeficiency virus (HIV)-induced immune deficiency remains unknown. A three-phase T cell reconstitution was demonstrated after HAART, with: (i) an early rise of memory CD4(+) cells, (ii) a reduction in T cell activation correlated to the decreasing retroviral activity together with an improved CD4(+) T cell reactivity to recall antigens, and (iii) a late rise of "naive" CD4(+) lymphocytes while CD8(+) T cells declined, however, without complete normalization of these parameters. Thus, decreasing the HIV load can reverse HIV-driven activation and CD4(+) T cell defects in advanced HIV-infected patients. PMID- 9204896 TI - Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. AB - Angiogenesis is thought to depend on a precise balance of positive and negative regulation. Angiopoietin-1 (Ang1) is an angiogenic factor that signals through the endothelial cell-specific Tie2 receptor tyrosine kinase. Like vascular endothelial growth factor, Ang1 is essential for normal vascular development in the mouse. An Ang1 relative, termed angiopoietin-2 (Ang2), was identified by homology screening and shown to be a naturally occurring antagonist for Ang1 and Tie2. Transgenic overexpression of Ang2 disrupts blood vessel formation in the mouse embryo. In adult mice and humans, Ang2 is expressed only at sites of vascular remodeling. Natural antagonists for vertebrate receptor tyrosine kinases are atypical; thus, the discovery of a negative regulator acting on Tie2 emphasizes the need for exquisite regulation of this angiogenic receptor system. PMID- 9204897 TI - Crystal structure of the cytochrome bc1 complex from bovine heart mitochondria. AB - On the basis of x-ray diffraction data to a resolution of 2.9 angstroms, atomic models of most protein components of the bovine cytochrome bc1 complex were built, including core 1, core 2, cytochrome b, subunit 6, subunit 7, a carboxyl terminal fragment of cytochrome c1, and an amino-terminal fragment of the iron sulfur protein. The positions of the four iron centers within the bc1 complex and the binding sites of the two specific respiratory inhibitors antimycin A and myxothiazol were identified. The membrane-spanning region of each bc1 complex monomer consists of 13 transmembrane helices, eight of which belong to cytochrome b. Closely interacting monomers are arranged as symmetric dimers and form cavities through which the inhibitor binding pockets can be accessed. The proteins core 1 and core 2 are structurally similar to each other and consist of two domains of roughly equal size and identical folding topology. PMID- 9204898 TI - X-ray and molecular emission from the nearest region of recent star formation. AB - The isolated, young, sunlike star TW Hya and four other young stars in its vicinity are strong x-ray sources. Their similar x-ray and optical properties indicate that the stars make up a physical association that is on the order of 20 million years old and that lies between about 40 and 60 parsecs (between about 130 and 200 light years) from Earth. TW Hya itself displays circumstellar CO, HCN, CN, and HCO+ emission. These molecules probably orbit the star in a solar system-sized disk viewed more or less face-on, whereas the star is likely viewed pole-on. Being at least three times closer to Earth than any well-studied region of star formation, the TW Hya Association serves as a test-bed for the study of x ray emission from young stars and the formation of planetary systems around sunlike stars. PMID- 9204905 TI - Identification of maize histone deacetylase HD2 as an acidic nucleolar phosphoprotein. AB - The steady state of histone acetylation is established and maintained by multiple histone acetyltransferases and deacetylases, and this steady state affects chromatin structure and function. The identification of a maize complementary DNA encoding the chromatin-bound deacetylase HD2 is reported. This protein was not homologous to the yeast RPD3 transcriptional regulator. It was expressed throughout embryo germination in correlation with the proliferative activity of cells. Antibodies against recombinant HD2-p39 immunoprecipitated the native enzyme complex, which was composed of phosphorylated p39 subunits. Immunofluorescence microscopy and sequence homologies suggested nucleolar localization. HD2 is an acidic nucleolar phosphoprotein that might regulate ribosomal chromatin structure and function. PMID- 9204906 TI - Cellular differentiation regulated by gibberellin in the Arabidopsis thaliana pickle mutant. AB - The plant growth regulator gibberellin (GA) has a profound effect on shoot development and promotes developmental transitions such as flowering. Little is known about any analogous effect GA might have on root development. In a screen for mutants, Arabidopsis plants carrying a mutation designated pickle (pkl) were isolated in which the primary root meristem retained characteristics of embryonic tissue. Expression of this aberrant differentiation state was suppressed by GA. Root tissue from plants carrying the pkl mutation spontaneously regenerated new embryos and plants. PMID- 9204907 TI - Evolution of a strain of CJD that induces BSE-like plaques. AB - Bovine spongiform encephalopathy (BSE) has become a public health issue because a recently evolved BSE agent has infected people, yielding an unusual form of Creutzfeld-Jakob disease (CJD). A new CJD agent that provokes similar amyloid plaques and cerebellar pathology was serially propagated. First-passage rats showed obvious clinical signs and activated microglia but had negligible PrP-res (the more protease-resistant form of host PrP) or cerebellar lesions. Microglia and astrocytes may participate in strain selection because the agent evolved, stabilized, and reproducibly provoked BSE-like disease in subsequent passages. Early vacuolar change involving activated microglia and astrocytes preceded significant PrP-res accumulation by more than 50 days. These studies reveal several inflammatory host reactions to an exogenous agent. PMID- 9204908 TI - Phosphorylation of the translational repressor PHAS-I by the mammalian target of rapamycin. AB - The immunosuppressant rapamycin interferes with G1-phase progression in lymphoid and other cell types by inhibiting the function of the mammalian target of rapamycin (mTOR). mTOR was determined to be a terminal kinase in a signaling pathway that couples mitogenic stimulation to the phosphorylation of the eukaryotic initiation factor (eIF)-4E-binding protein, PHAS-I. The rapamycin sensitive protein kinase activity of mTOR was required for phosphorylation of PHAS-I in insulin-stimulated human embryonic kidney cells. mTOR phosphorylated PHAS-I on serine and threonine residues in vitro, and these modifications inhibited the binding of PHAS-I to eIF-4E. These studies define a role for mTOR in translational control and offer further insights into the mechanism whereby rapamycin inhibits G1-phase progression in mammalian cells. PMID- 9204909 TI - Immunogold localization of the dopamine transporter: an ultrastructural study of the rat ventral tegmental area. AB - The dopamine transporter (DAT) plays an important role in the plasmalemmal reuptake of dopamine and, thus, in the termination of normal dopaminergic neurotransmission. DAT is also a major binding site for cocaine and other stimulants, the psychoactive effects of which are associated primarily with the inhibition of dopamine reuptake within mesocorticolimbic dopaminergic neurons. We used electron microscopy with an anti-peptide antiserum directed against the N terminal domain of DAT to determine the subcellular localization of this transporter in the rat ventral tegmental area (VTA), the region that contains the cell bodies and dendrites of these dopaminergic neurons. We show that in the VTA, almost 95% of the DAT immunogold-labeled profiles are neuronal perikarya and dendrites, and the remainder are unmyelinated axons. Within perikarya and large proximal dendrites, almost all of the DAT immunogold particles are associated with intracellular membranes, including saccules of Golgi and cytoplasmic tubulovesicles. In contrast, within medium- to small-diameter dendrites and unmyelinated axons, most of the DAT gold particles are located on plasma membranes. In dually labeled tissue, peroxidase reaction product for the catecholamine-synthesizing enzyme tyrosine hydroxylase is present in DAT immunoreactive profiles. These findings suggest that intermediate and distal dendrites are both the primary sites of dopamine reuptake and the principal targets of cocaine and related psychostimulants within dopaminergic neurons in the VTA. PMID- 9204910 TI - Alpha-conotoxin MII blocks nicotine-stimulated dopamine release in rat striatal synaptosomes. AB - Activation of presynaptic nicotinic acetylcholine receptors (nAChRs) can induce the release of neurotransmitters such as dopamine and norepinephrine in the CNS. Accumulating evidence suggests that distinct nAChR subtypes are involved; however, it has been difficult to determine the subunit composition of these receptors, in part because of the lack of a sufficient variety of selective nAChR ligands. We present experimental data that at least two different nAChR complexes are involved in dopamine release, one of which has an alpha3/beta2 subunit interface. The recently discovered peptide alpha-conotoxin MII is a potent and selective inhibitor of rat nAChRs containing an interface formed by alpha3 and beta2 subunits. We used this peptide to examine nicotine-stimulated release of dopamine from rat striatal synaptosomes and of norepinephrine from hippocampal synaptosomes. MII (100 nM) blocks 34-49% of the nicotine-stimulated dopamine release, but not dopamine release evoked by elevated [K+]. Furthermore, two peptides structurally related to alpha-conotoxin MII, namely alpha-conotoxin MI (selective for alpha1beta1gammadelta nAChRs) and alpha-conotoxin ImI (selective for alpha7-containing nAChRs), have no effect on nicotine-stimulated dopamine release. The results indicate that one third to half of the dopamine release in the striatal preparation is mediated by nAChRs with an alpha3/beta2 subunit interface. In contrast, 30 min, i.e., manifesting long-term depression (LTD). The latter was switched into a short-term depression lasting 15-25 min after pharmacological blockade of NMDA receptor channels with D-APV or chelation of intracellular calcium ions with EGTA, whereas the accommodation phase was unaffected. Application of an AMPA receptor anti-desensitization agent cyclothiazide abolished the LTD, but not the accommodation response. These results suggest the presence of separate postsynaptic sites for the induction of LTD and accommodation, one being sensitive to cyclothiazide, whereas the other is not. Moreover, the maintenance of LTD is dependent on the level of intracellular Ca2+. These phasic and long-term synaptic plasticity in the nucleus tractus solitarius may play a role in the homeostatic regulation of cardiorespiratory functions. PMID- 9204920 TI - Postsynaptic inhibitors of calcium/calmodulin-dependent protein kinase type II block induction but not maintenance of pairing-induced long-term potentiation. AB - The role of postsynaptic kinases in the induction and maintenance of long-term potentiation (LTP) was studied in the CA1 region of the rat hippocampal slice. A peptide inhibitor for the catalytic domain of calcium/calmodulin-dependent protein kinase type II (CaM-kinase) was applied through a perfused patch pipette. The inhibitor completely blocked both the short-term potentiation and LTP induced by a pairing protocol. This indicates that the kinase or kinases affected by the peptide are downstream from depolarization in the LTP cascade. The ability to block LTP required that measures be taken to interfere with degradation of the peptide kinase inhibitor by endogenous proteases; either addition of protease inhibitors or modifications of the peptide itself greatly enhanced the effectiveness of the peptide. Protease inhibitors by themselves or control peptide did not block LTP induction. To study the effect of kinase inhibitor on LTP maintenance, we induced LTP in one pathway. Subsequent introduction of the kinase inhibitor blocked the induction of LTP in a second pathway, but it did not affect maintenance of LTP in the first. The implications for the role of kinases in LTP maintenance are discussed. PMID- 9204921 TI - A long-lasting calcium-activated nonselective cationic current is generated by synaptic stimulation or exogenous activation of group I metabotropic glutamate receptors in CA1 pyramidal neurons. AB - We have shown previously that a selective metabotropic glutamate receptor (mGluR) agonist, 1S,3R-1-aminocyclo-pentane-1, 3-dicarboxylate (1S,3R-ACPD), evokes an inward current in CA1 pyramidal neurons of rat hippocampal slices in the presence of K+ channel blockers (). This current has been characterized as a Ca2+ activated nonselective cationic (CAN) current. Using whole-cell patch-clamp recordings and intracellular dialysis, we now have identified the mGluR subtype and the mechanisms underlying the CAN current (ICAN) and report for the first time the presence of a synaptic ICAN in the mammalian CNS. First, we have shown pharmacologically that activation of ICAN by 1S,3R-ACPD involves the group I mGluRs (and not the groups II and III) and a G-protein-dependent process. We also report that ICAN is modulated by the divalent cations (Mg2+, Cd2+, and Zn2+). Second, we have isolated a slow synaptic inward current evoked by a high frequency stimulation in the presence of K+ channel blockers, ionotropic glutamate, and GABAA receptor antagonists. This current shows similar properties to the exogenously evoked ICAN: its reversal potential is close to the reversal potential of the 1S, 3R-ACPD-evoked ICAN, and it is G-protein- and Ca2+ dependent. Because the amplitude and duration of ICAN increased in the presence of a glutamate uptake blocker, we suggest that this synaptic current is generated via the activation of mGluRs. We propose that the synaptic ICAN, activated by a brief tetanic stimulation and leading to a long-lasting inward current, may be involved in neuronal plasticity and synchronized network-driven oscillations. PMID- 9204922 TI - A novel type of GABAergic interneuron connecting the input and the output regions of the hippocampus. AB - The main excitatory pathway of the hippocampal formation is controlled by a network of morphologically distinct populations of GABAergic interneurons. Here we describe a novel type of GABAergic interneuron located in the outer molecular layer (OML) of the rat dentate gyrus with a long-range forward projection from the dentate gyrus to the subiculum across the hippocampal fissure. OML interneurons were recorded in hippocampal slices by using the whole-cell patch clamp configuration. During recording, cells were filled with biocytin for subsequent light and electron microscopic analysis. Neurons projecting to the subiculum were distributed throughout the entire OML. They had round or ovoid somata and a multipolar dendritic morphology. Two axonal domains could be distinguished: an extensive, tangential distribution within the OML and a long range vertical and tangential projection to layer 1 and stratum pyramidale of the subiculum. Symmetric synaptic contacts were established by these interneurons on dendritic shafts in the OML and subiculum. OML interneurons were characterized physiologically by short action potential duration and marked afterhyperpolarization that followed the spike. On sustained current injection, they generated high-frequency (up to 130 Hz, 34 degrees C) trains of action potentials with only little adaptation. In situ hybridization and single-cell RT PCR analysis for GAD67 mRNA confirmed the GABAergic nature of OML interneurons. GABAergic interneurons in the OML projecting to the subiculum connect the input and output regions of the hippocampus. Hence, they could mediate long-range feed forward inhibition and may participate in an oscillating cross-regional interneuron network that may synchronize the activity of spatially distributed principal neurons in the dentate gyrus and the subiculum. PMID- 9204923 TI - Neuronal and glial apoptosis after traumatic spinal cord injury. AB - Cell death was examined by studying the spinal cords of rats subjected to traumatic insults of mild to moderate severity. Within minutes after mild weight drop impact (a 10 gm weight falling 6.25 mm), neurons in the immediate impact area showed a loss of cytoplasmic Nissl substances. Over the next 7 d, this lesion area expanded and cavitated. Terminal deoxynucleotidyl transferase (TdT) mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL)-positive neurons were noted primarily restricted to the gross lesion area 4-24 hr after injury, with a maximum presence at 8 hr after injury. TUNEL-positive glia were present at all stages studied between 4 hr and 14 d, with a maximum presence within the lesion area 24 hr after injury. However 7 d after injury, a second wave of TUNEL-positive glial cells was noted in the white matter peripheral to the lesion and extending at least several millimeters away from the lesion center. The suggestion of apoptosis was supported by electron microscopy, as well as by nuclear staining with Hoechst 33342 dye, and by examination of DNA prepared from the lesion site. Furthermore, repeated intraperitoneal injections of cycloheximide, beginning immediately after a 12.5 mm weight drop insult, produced a substantial reduction in histological evidence of cord damage and in motor dysfunction assessed 4 weeks later. Present data support the hypothesis that apoptosis dependent on active protein synthesis contributes to the neuronal and glial cell death, as well as to the neurological dysfunction, induced by mild-to moderate severity traumatic insults to the rat spinal cord. PMID- 9204924 TI - Absence of a persistently elevated 37 kDa fos-related antigen and AP-1-like DNA binding activity in the brains of kainic acid-treated fosB null mice. AB - Chronic stimulation of the nervous system or acute administration of kainic acid results in a persistent increase in AP-1-like DNA-binding activity in the brain. However, the composition and function of these AP-1 complexes remain controversial. By comparing wild-type and fosB-null mice treated with kainic acid, we establish that the complexes comprise JunD in association with an approximately 37 kDa Delta-FosB species. Delta-FosB was expressed persistently in neurons in many areas of the CNS, even though fosB mRNA only increased transiently. This implies that the 37 kDa protein is very stable. fosB-/- mice are predisposed to seizures. Therefore, the chronic expression of Delta-FosB elicited by kainic acid seizures may be indicative of a compensatory/protective role in the pathophysiology of epilepsy. PMID- 9204925 TI - Visualization of the distribution of autophosphorylated calcium/calmodulin dependent protein kinase II after tetanic stimulation in the CA1 area of the hippocampus. AB - Autophosphorylation of calcium/calmodulin-dependent protein kinase II (CaMKII) at threonine-286 produces Ca2+-independent kinase activity and has been proposed to be involved in induction of long-term potentiation by tetanic stimulation in the hippocampus. We have used an immunocytochemical method to visualize and quantify the pattern of autophosphorylation of CaMKII in hippocampal slices after tetanization of the Schaffer collateral pathway. Thirty minutes after tetanic stimulation, autophosphorylated CaM kinase II (P-CaMKII) is significantly increased in area CA1 both in apical dendrites and in pyramidal cell somas. In apical dendrites, this increase is accompanied by an equally significant increase in staining for nonphosphorylated CaM kinase II. Thus, the increase in P-CaMKII appears to be secondary to an increase in the total amount of CaMKII. In neuronal somas, however, the increase in P-CaMKII is not accompanied by an increase in the total amount of CaMKII. We suggest that tetanic stimulation of the Schaffer collateral pathway may induce new synthesis of CaMKII molecules in the apical dendrites, which contain mRNA encoding its alpha-subunit. In neuronal somas, however, tetanic stimulation appears to result in long-lasting increases in P CaMKII independent of an increase in the total amount of CaMKII. Our findings are consistent with a role for autophosphorylation of CaMKII in the induction and/or maintenance of long-term potentiation, but they indicate that the effects of tetanus on the kinase and its activity are not confined to synapses and may involve induction of new synthesis of kinase in dendrites as well as increases in the level of autophosphorylated kinase. PMID- 9204926 TI - Postnatal development of membrane properties and delta oscillations in thalamocortical neurons of the cat dorsal lateral geniculate nucleus. AB - The development of membrane properties, firing patterns, and delta oscillations in neurons of the cat dorsal lateral geniculate nucleus (dLGN) was investigated in vitro during the first 7 postnatal weeks. Compared with adult neurons, the resting membrane potential was more depolarized at postnatal days 1-9 (P1-P9), the input resistance was higher at P1-P7, and action potentials had a higher threshold and a smaller amplitude at P1-P3 and a longer duration at P1-P9. At P1 P3 trains longer than 200 msec were rarely observed, and trains with more than three action potentials were only present in 41% of the neurons, whereas at P1-P7 the normalized slope of the instantaneous frequencies at the first five interspike intervals was smaller than in the adult. A long-lasting (up to 6 sec) afterhyperpolarization followed a short train of action potentials in 88 and 30% of neurons at P1-P3 and P30-P32, respectively, but it was rarely observed in the adult. The low-threshold Ca2+ potential could evoke a burst of action potentials since P1. However, at P1-P7 the number of action potentials per burst was smaller (range, one to five), and at P1-P9 their maximum instantaneous frequency was lower (<190 Hz) than in the adult (range, six to eight, and 344 Hz, respectively). No delta oscillations were observed until P17, and their frequency (0.36 Hz) was lower than that in the adult (1.8 Hz). The percentage of neurons displaying delta oscillations and their frequency reached adult values by the end of the seventh postnatal week, i.e., well after the maturation of the membrane properties and firing patterns (second postnatal week). In conclusion, the maturation of the electrophysiological properties of thalamocortical neurons in the cat dLGN is completed later than the retinogeniculate axon segregation (Shatz CJ, 1983), and the immaturity of the oscillatory, and not of the burst-firing, activity is a limiting factor in the development of delta waves. PMID- 9204927 TI - The trkA receptor mediates growth cone turning toward a localized source of nerve growth factor. AB - We have developed an in vitro system for studying the interaction of chick dorsal root ganglion neuronal growth cones with a localized source of nerve growth factor (NGF) covalently conjugated to polystyrene beads. Growth cones rapidly turned and migrated under NGF-coated beads in a process that involved the initial formation of persistent contact with a bead, followed by directed flow of cytoplasm toward the point of contact. A role for the local activation of the high-affinity NGF receptor trkA was suggested by a strong inhibition of the turning response by (1) the addition of an antibody against the extracellular portion of trkA, (2) the elevation of the background concentration of NGF to saturate trkA, or (3) the presence of a concentration of the drug K252a that inhibits trkA activation. NGF binding to the pan-neurotrophin receptor p75 is also involved but is not required for turning. These data show a new role for both the trkA and the p75 receptors: the mediation of local events in the guidance of nerve growth cones. PMID- 9204928 TI - Nerve growth factor stimulates the accumulation of beta1 integrin at the tips of filopodia in the growth cones of sympathetic neurons. AB - Addition of nerve growth factor (NGF) to sympathetic neurons that have been starved of it causes a rapid induction of growth cone motility and the resumption of neurite growth. Using immunofluorescence staining, we show that within 10 min, NGF stimulated the accumulation of dense aggregates of beta1 integrin [a receptor for extracellular matrix (ECM) proteins] at most of the tips of either newly extended or preexisting filopodia. This effect occurred in the absence of ECM proteins and in the presence of 1 mg/ml Arg-Gly-Asp-Ser peptide, which blocks ECM binding to integrin, indicating that occupation of the integrin receptor is not necessary for tip localization. In fact, addition of either laminin or fibronectin caused a rapid withdrawal of beta1 integrin aggregates from filopodial tips at a rate comparable to that of the rearward flow of actin filaments in the periphery of the growth cone. Surface labeling of the extracellular domain of beta1 integrin while aggregated at the tips of filopodia or withdrawing in response to ECM proteins showed that the receptor is positioned within the membrane. The drug butanedione monoxime, an inhibitor of myosins, blocked the accumulation of beta1 integrin at the tips of filopodia without inhibiting the formation of filo-podia, suggesting the involvement of a myosin motor in beta1 integrin transport. These results provide the first evidence of NGF-mediated accumulation of ECM receptors to sensory elements of the growth cone and suggest one mechanism whereby soluble and substrate-bound cues coordinate to produce directed neurite growth. PMID- 9204929 TI - Immunohistochemical localization of netrin-1 in the embryonic chick nervous system. AB - Netrin-1 has profound in vitro effects on the growth properties of vertebrate embryonic axons. In addition, netrin-1 mRNA is found in the floor plate of the embryonic nervous system, an intermediate target of many axons, including commissural axons that are affected by netrin-1 in vitro. Moreover, genetic studies of netrin-1 homologs in Caenorhabditis elegans and Drosophila implicate these proteins in commissure formation. We raised polyclonal antisera that recognize chick netrin-1 in fixed tissue sections. The antisera were used to immunohistochemically map netrin-1 in the embryonic spinal cord, brain, and retina. The relationship between netrin-1 localization and the growth of pioneering axons suggests roles for netrin-1 in the regulation of circumferential, commissural, and longitudinal axon growth in the spinal cord and brain. The data also suggest that the primary or sole effect of netrin-1 on pioneering spinal cord commissural axons is haptotactic. Furthermore, the pattern of netrin-1 localization raises the possibility that this protein helps mediate neuronal migration in the spinal cord, brain, and retina. PMID- 9204930 TI - Functional specificity of long-range intrinsic and interhemispheric connections in the visual cortex of strabismic cats. AB - The development of both long-range intracortical and interhemispheric connections depends on visual experience. Previous experiments showed that in strabismic but not in normal cats, clustered horizontal axon projections preferentially connect cell groups activated by the same eye. This indicates that there is selective stabilization of fibers between neurons exhibiting correlated activity. Extending these experiments, we investigated in strabismic cats: (1) whether tangential connections remain confined to columns of similar orientation preference within the subsystems of left and right eye domains; and (2) whether callosal connections also extend predominantly between neurons activated by the same eye and preferring similar orientations. To this end, we analyzed in strabismic cats the topographic relationships between orientation preference domains and both intrinsic and callosal connections of area 17. Red and green latex microspheres were injected into monocular iso-orientation domains identified by optical imaging of intrinsic signals. Additionally, domains sharing the ocular dominance and orientation preference of the neurons at the injection sites were visualized by 2-deoxyglucose (2-DG) autoradiography. Quantitative analysis revealed that 56% of the retrogradely labeled cells within the injected area 17 and 60% of the transcallosally labeled neurons were located in the 2-DG-labeled iso-orientation domains. This indicates: (1) that strabismus does not interfere with the tendency of long-range horizontal fibers to link predominantly neurons of similar orientation preference; and (2) that the selection mechanisms for the stabilization of callosal connections are similar to those that are responsible for the specification of the tangential intrinsic connections. PMID- 9204931 TI - HNMP-1: a novel hematopoietic and neural membrane protein differentially regulated in neural development and injury. AB - The hnmp-1 (hematopoietic neural membrane protein) gene encodes a protein with striking similarity to the tetra-transmembrane-spanning protein encoded by pmp22. hnmp-1 was cloned from an elutriated human monocyte library and is expressed in various human hematopoietic and lymphoid lineages as well as adult mouse spleen and thymus. In the mouse nervous system, HNMP-1 mRNA is temporally expressed by Schwann cells during sciatic nerve myelination. Dorsal root ganglia sensory and spinal cord alpha-motoneurons acquire HNMP-1 protein selectively throughout development. In the fiber tracts of the spinal cord and in sciatic nerve, HNMP-1 protein is axon-associated. Additionally a rapid and sustained level of HNMP-1 expression is observed in response to acute PNS injury. HNMP-1 is constituitively induced in sciatic nerve of Trembler J mice, which are mutant for pmp22 and have a demyelinating/hypomyelinating phenotype. The expression pattern of HNMP-1 suggests a possible role for this molecule during active myelination. PMID- 9204932 TI - Effects of regional anesthesia on phantom limb pain are mirrored in changes in cortical reorganization. AB - The causes underlying phantom limb pain are still unknown. Recent studies on the consequences of nervous system damage in animals and humans reported substantial reorganization of primary somatosensory cortex subsequent to amputation, and one study showed that cortical reorganization is positively correlated with phantom limb pain. This paper examined the hypothesis of a functional relationship between cortical reorganization and phantom limb pain. Neuroelectric source imaging was used to determine changes in cortical reorganization in somatosensory cortex after anesthesia of an amputation stump produced by brachial plexus blockade in six phantom limb pain patients and four pain-free amputees. Three of six phantom limb subjects experienced a virtual elimination of current phantom pain attributable to anesthesia (mean change: 3.8 on an 11-point scale; Z = 1.83; p < 0.05) that was mirrored by a very rapid elimination of cortical reorganization in somatosensory cortex (change = 19.8 mm; t(2) = 5.60; p < 0.05). Cortical reorganization remained unchanged (mean change = 1.6 mm) in three phantom limb pain amputees whose pain was not reduced by brachial plexus blockade and in the phantom pain-free amputation controls. These findings suggest that cortical reorganization and phantom limb pain might have a causal relationship. Methods designed to alter cortical reorganization should be examined for their efficacy in the treatment of phantom limb pain. PMID- 9204933 TI - GABAergic neurons in barrel cortex show strong, whisker-dependent metabolic activation during normal behavior. AB - Electrophysiological data from the rodent whisker/barrel cortex indicate that GABAergic, presumed inhibitory, neurons respond more vigorously to stimulation than glutamatergic, presumed excitatory, cells. However, these data represent very small neuronal samples in restrained, anesthetized, or narcotized animals or in cortical slices. Histochemical data from primate visual cortex, stained for the mitochondrial enzyme cytochrome oxidase (CO) and for GABA, show that GABAergic neurons are more highly reactive for CO than glutamatergic cells, indicating that inhibitory neurons are chronically more active than excitatory neurons but leaving doubt about the short-term stimulus dependence of this activation. Taken together, these results suggest that highly active inhibitory neurons powerfully influence relatively inactive excitatory cells but do not demonstrate directly the relative activities of excitatory and inhibitory neurons in the cortex during normal behavior. We used a novel double-labeling technique to approach the issue of excitatory and inhibitory neuronal activation during behavior. Our technique combines high-resolution 2-deoxyglucose (2DG), immunohistochemical staining for neurotransmitter-specific antibodies, and automated image analysis to collect the data. We find that putative inhibitory neurons in barrel cortex of behaving animals are, on average, much more heavily 2DG-labeled than presumed excitatory cells, a pattern not seen in animals anesthetized at the time of 2DG injection. This metabolic activation is dependent specifically on sensory inputs from the whiskers, because acute trimming of most whiskers greatly reduces 2DG labeling in both cell classes in columns corresponding to trimmed whiskers. Our results provide confirmation of the active GABAergic cell hypothesis suggested by CO and single-unit data. We conclude that strong activation of inhibitory cortical neurons must confer selective advantages that compensate for its inherent energy inefficiency. PMID- 9204934 TI - Distributed neural systems underlying the timing of movements. AB - Timing is essential to the execution of skilled movements, yet our knowledge of the neural systems underlying timekeeping operations is limited. Using whole brain functional magnetic resonance imaging, subjects were imaged while tapping with their right index finger in synchrony with tones that were separated by constant intervals [Synchronization (S)], followed by tapping without the benefit of an auditory cue [Continuation (C)]. Two control conditions followed in which subjects listened to tones and then made pitch discriminations (D). Both the S and the C conditions produced equivalent activation within the left sensorimotor cortex, the right cerebellum (dorsal dentate nucleus), and the right superior temporal gyrus (STG). Only the C condition produced activation of a medial premotor system, including the caudal supplementary motor area (SMA), the left putamen, and the left ventrolateral thalamus. The C condition also activated a region within the right inferior frontal gyrus (IFG), which is functionally interconnected with auditory cortex. Both control conditions produced bilateral activation of the STG, and the D condition also activated the rostral SMA. These results suggest that the internal generation of precisely timed movements is dependent on three interrelated neural systems, one that is involved in explicit timing (putamen, ventrolateral thalamus, SMA), one that mediates auditory sensory memory (IFG, STG), and another that is involved in sensorimotor processing (dorsal dentate nucleus, sensorimotor cortex). PMID- 9204935 TI - Cells in laminae III and IV of the rat spinal cord that possess the neurokinin-1 receptor and have dorsally directed dendrites receive a major synaptic input from tachykinin-containing primary afferents. AB - Many neurons with cell bodies in laminae III or IV of the spinal dorsal horn possess the neurokinin 1 receptor and have dorsal dendrites that arborize in the superficial dorsal horn. We have performed a confocal microscopic study to determine whether these cells receive inputs from substance P-containing primary afferents. All neurons of this type received contacts from substance P immunoreactive axons, and in most cases the contacts onto dorsal dendrites were very numerous. A great majority (90-100%) of substance P-immunoreactive varicosities in contact with these cells were also immunoreactive with antibody to calcitonin gene-related peptide, indicating that they were of primary afferent origin. The density of contacts from substance P-immunoreactive varicosities onto these cells was significantly higher than that seen on cholinergic neurons in lamina III (which do not possess the receptor). Electron microscopy revealed that synapses were present at points of contact between substance P-immunoreactive boutons and dorsal dendrites of cells with the neurokinin 1 receptor. Some cells of this type belong to the spinothalamic tract, and we therefore examined neurons with cell bodies in laminae III or IV that possessed the neurokinin 1 receptor and were labeled retrogradely after thalamic injection of cholera toxin B subunit. These cells also received contacts from substance P-immunoreactive axons on their dorsal dendrites. The results of this study indicate that neurons of this type are a major target for substance P-containing primary afferents. PMID- 9204936 TI - Expression of LIM protein genes Lmo1, Lmo2, and Lmo3 in adult mouse hippocampus and other forebrain regions: differential regulation by seizure activity. AB - The LIM domain is a zinc-binding amino acid motif that characterizes various proteins which function in protein-protein interactions and transcriptional regulation. Expression patterns of several LIM protein genes are compatible with roles in vertebrate CNS development, but little is known about the expression, regulation, or function of LIM proteins in the mature CNS. Lmo1, Lmo2, and Lmo3 are LIM-only genes originally identified as putative oncogenes that have been implicated in the control of cell differentiation and are active during CNS development. Using in situ hybridization for mRNA and immunohistochemical detection of reporter protein expression in transgenic mice, we found that Lmo1, Lmo2, and Lmo3 show individually unique but partially overlapping patterns of expression in several regions of the adult mouse forebrain, including hippocampus, caudate putamen, medial habenula, thalamus, amygdala, olfactory bulb, hypothalamus, and cerebral cortex. In the hippocampal formation, Lmo1, Lmo2, and Lmo3 show different combinatorial patterns of expression levels in CA pyramidal and dentate granule neurons, and Lmo1 is present in topographically restricted subpopulations of astrocytes. Kainic acid-induced limbic seizures differentially regulated Lmo1, Lmo2, and Lmo3 mRNA levels in hippocampal pyramidal and granule neurons, such that Lmo1 mRNA increased, whereas Lmo2 and Lmo3 mRNAs decreased significantly, with maximal changes at 6 hr after seizure onset and return to baseline by 24 hr. These findings show that Lmo1, Lmo2, and Lmo3 continue to be expressed in the adult mammalian CNS in a cell type-specific manner, are differentially regulated by neuronal activity, and may thus be involved in cell phenotype-specific regulatory functions. PMID- 9204937 TI - Cellular delivery of neurotrophin-3 promotes corticospinal axonal growth and partial functional recovery after spinal cord injury. AB - The injured adult mammalian spinal cord shows little spontaneous recovery after injury. In the present study, the contribution of projections in the dorsal half of the spinal cord to functional loss after adult spinal cord injury was examined, together with the effects of transgenic cellular delivery of neurotrophin-3 (NT-3) on morphological and functional disturbances. Adult rats underwent bilateral dorsal column spinal cord lesions that remove the dorsal corticospinal projections or underwent more extensive resections of the entire dorsal spinal cord bilaterally that remove corticospinal, rubrospinal, and cerulospinal projections. Long-lasting functional deficits were observed on a motor grid task requiring detailed integration of sensorimotor skills, but only in animals with dorsal hemisection lesions as opposed to dorsal column lesions. Syngenic primary rat fibroblasts genetically modified to produce NT-3 were then grafted to acute spinal cord dorsal hemisection lesion cavities. Up to 3 months later, significant partial functional recovery occurred in NT-3-grafted animals together with a significant increase in corticospinal axon growth at and distal to the injury site. These findings indicate that (1) several spinal pathways contribute to loss of motor function after spinal cord injury, (2) NT-3 is a neurotrophic factor for the injured corticospinal projection, and (3) functional deficits are partially ameliorated by local cellular delivery of NT-3. Lesions of the corticospinal projection may be necessary, but insufficient in isolation, to cause sensorimotor dysfunction after spinal cord injury in the rat. PMID- 9204938 TI - Adrenergic receptors in Alzheimer's disease brain: selective increases in the cerebella of aggressive patients. AB - In this study, the distribution and concentration of beta1, beta2, and alpha2 adrenergic receptors were examined in the frontal cortex, hypothalamus, and cerebellum of Alzheimer's disease (AD) and age-matched control human brains by receptor autoradiography. The purpose of this study was to detect changes in adrenergic receptor concentrations in key areas of the brain known to affect behavior. For these studies, [125I]iodopindolol ([125I]IPIN) was used to visualize total beta adrenergic sites (with ICI-89,406 and ICI-118, 551 as subtype-selective antagonists to visualize beta2 and beta1 receptors, respectively). [3H]UK-14,304 was used to localize the alpha2 sites. Essentially no significant difference in adrenergic receptor concentration was found between total AD cases taken together and control patients. It was found, however, that there were important distinctions within the AD group when cases were subdivided according to the presence or absence of aggression, agitation, and disruptive behavior. Aggressive AD patients had markedly increased (by approximately 70%) concentrations of alpha2 receptors in the cerebellar cortex compared with nonaggressive patients with similar levels of cognitive deficit. The levels of cerebellar alpha2 receptors in aggressive AD patients were slightly above the healthy elderly controls, suggesting that these receptors are preserved and perhaps increased in this subgroup of AD. beta1 And beta2 adrenergic receptors of the cerebellar cortex showed smaller but significant ( approximately 25%) increases in concentration in aggressive AD subjects versus both nonaggressive AD patients and controls. No significant differences were found in adrenergic receptor concentrations within the frontal cortex or hypothalamus. These results point out the importance of distinguishing behavioral subgroups of AD when looking for specific neurochemical changes. These autoradiographic results may reflect the importance of the cerebellum in behavioral control. PMID- 9204940 TI - Serotonin 5-HT1A receptors modulate hippocampal reactivity to afferent stimulation. AB - Hippocampal dentate gyrus reactivity to perforant path (PP) stimulation in the anesthetized rat was enhanced after systemic administration of the serotonin releasing drug fenfluramine (FFA). This effect of FFA was mimicked by local application of the drug via the recording pipette, indicating that the effect of FFA is mediated by release of serotonin from intrahippocampal serotonergic terminals. The 5-HT1a antagonist NAN-190 and the 5-HT1b agonist CGS-12066-B, applied both systemically and locally, blocked the effect of FFA. This blocking action was not shared by the 5-HT2-4 receptor agonists or antagonists tested. The 5-HT1a receptor agonist 8-OH-DPAT, applied systemically, caused a marked reduction in population spike responses to PP stimulation, whereas an opposite effect was produced by local application of this drug. The effect of peripheral application of 8-OH-DPAT was blocked by depletion of serotonin. The local effect of FFA was blocked by a reducing neurotransmitter release with a pipette containing 10 mM Mg2+. Finally, local application of the GABA antagonist picrotoxin also enhanced population spike response to PP stimulation, and the effects of picrotoxin and FFA occluded. These results indicate that serotonin released from terminals in the hippocampus activates a 5-HT1a receptor on interneurons that suppresses their activity and thus enhances dentate granular cell population spike response to PP stimulation. PMID- 9204939 TI - Repeated generalized seizures induce time-dependent changes in the behavioral seizure response independent of continued seizure induction. AB - This study examined both the acute and long-lasting changes in seizure susceptibility that occur in response to the repeated induction of generalized seizure activity. Daily flurothyl-induced generalized clonic seizures resulted in a progressive decrease in both the generalized seizure threshold and the latency to the first myoclonic jerk. The threshold reduction was significant as early as the second trial and was maximal by trial 5. However, a minimum of eight seizures was necessary for the maximal reduction to be long-lasting. The present study also examined the effects of the number of seizures and the duration of the stimulation-free interval on the type of generalized seizure expressed. During the induction phase of the experiment, only generalized clonic seizures ("forebrain seizures") were expressed. If, however, the animal was retested after a 1, 2, 3, or 4 week stimulation-free interval, a progressive increase in both the proportion of animals expressing "brainstem seizure" behaviors and the median seizure score was observed. The progression of flurothyl-induced generalized seizure behaviors was significantly altered if (1) a minimum of eight generalized clonic seizures had been expressed, and (2) a minimum of a 2 week stimulation free interval followed. Fewer generalized clonic seizures failed to reliably produce changes in seizure phenotype, even after extended stimulus-free intervals. These data indicate that specific kindling processes are initiated during the interval of repeated seizure induction and evolve in the absence of continued seizure induction. Furthermore, these mechanisms of epileptogenesis were found to be manifest predominantly as a change in the seizure phenotype expressed and to proceed independent of changes in the generalized seizure threshold. PMID- 9204941 TI - Neonatal nonhandling and in utero prenatal stress reduce the density of NADPH diaphorase-reactive neurons in the fascia dentata and Ammon's horn of rats. AB - The density of nitric oxide (NO)-producing neurons in the fascia dentata and Ammon's horn was assessed in 6-month-old male rats using NADPH-diaphorase (NADPH d) histochemistry. Two separate experiments investigated whether (1) the complete absence of neonatal handling or (2) the administration of periodic prenatal stress could affect the expression and distribution of NADPH-d reactivity in the hippocampus, when compared with rats raised in normal standard laboratory conditions. Experiment 1 demonstrated that adult rats that received no handling during neonatal development (from birth to postnatal day 22) showed a very substantial reduction in NADPH-d-positive neurons per unit area throughout the entire hippocampus when compared with rats that received regular daily handling in this period. Quantitative analysis further revealed that this effect was significantly more pronounced in Ammon's horn than in the fascia dentata, and within Ammon's horn the dorsal region was selectively more affected. Experiment 2 showed that prenatal stress, which involved the administration of daily restraint stress to pregnant dams throughout the gestation period, also led to a reduction in NADPH-d reactivity in the hippocampus of the offspring of these dam when they reached adulthood. The present results suggest that behavioral manipulations in the early neonatal or prenatal period can significantly alter the neurodevelopment of the hippocampal NO system and these changes might be related to some of the behavioral abnormalities that emerge later in adulthood. PMID- 9204942 TI - Temporal dynamics of graded synaptic transmission in the lobster stomatogastric ganglion. AB - Synaptic transmission between neurons in the stomatogastric ganglion of the lobster Panulirus interruptus is a graded function of membrane potential, with a threshold for transmitter release in the range of -50 to -60 mV. We studied the dynamics of graded transmission between the lateral pyloric (LP) neuron and the pyloric dilator (PD) neurons after blocking action potential-mediated transmission with 0.1 microM tetrodotoxin. We compared the graded IPSPs (gIPSPs) from LP to PD neurons evoked by square pulse presynaptic depolarizations with those potentials evoked by realistic presynaptic waveforms of variable frequency, amplitude, and duty cycle. The gIPSP shows frequency-dependent synaptic depression. The recovery from depression is slow, and as a result, the gIPSP is depressed at normal pyloric network frequencies. Changes in the duration of the presynaptic depolarization produce nonintuitive changes in the amplitude and time course of the postsynaptic responses, which are again frequency-dependent. Taken together, these data demonstrate that the measurements of synaptic efficacy that are used to understand neural network function are best made using presynaptic waveforms and patterns of activity that mimic those in the functional network. PMID- 9204943 TI - Differential activation and desensitization of sensory neurons by resiniferatoxin. AB - Recently, with use of rat dorsal root ganglion (DRG) neurons we have been able to dissociate the binding affinities of vanilloids from their potencies to induce 45Ca uptake, which suggests the existence of distinct classes of the vanilloid receptor (). In the present study, we have demonstrated that the ultrapotent capsaicin analog resiniferatoxin (RTX) desensitized rat DRG neurons to the subsequent induction of 45Ca uptake by capsaicin and RTX with affinity and cooperativity similar to that found for [3H]RTX binding, contrasting with a approximately 10-fold weaker potency and lack of cooperativity to induce 45Ca uptake. Likewise, the competitive antagonist capsazepine inhibited RTX-induced desensitization with potency similar to that for inhibition of specific [3H]RTX binding, whereas the potency of capsazepine was approximately 10-fold higher for inhibiting RTX-induced 45Ca uptake. Finally, the noncompetitive antagonist ruthenium red inhibited both the RTX-induced desensitization and 45Ca uptake but showed approximately 60-fold selectivity for inhibiting RTX-induced desensitization. The RTX-induced desensitization was not associated with loss of specific [3H]RTX binding, suggesting lack of gross cell toxicity. In contrast to RTX, capsaicin caused desensitization with a potency corresponding to that for 45Ca uptake and did so in a noncooperative manner. Unlike the RTX-induced desensitization, the desensitization by capsaicin was blocked by ruthenium red only at doses that blocked 45Ca uptake and depended on external calcium. Our findings provide further support for the existence of vanilloid receptor subtypes on DRG neurons with distinct pharmacology and distinct patterns of desensitization. PMID- 9204944 TI - Calcium channel density and hippocampal cell death with age in long-term culture. AB - The expression of voltage-gated calcium (Ca2+) channel activity in brain cells is known to be important for several aspects of neuronal development. In addition, excessive Ca2+ influx has been linked clearly to neurotoxicity both in vivo and in vitro; however, the temporal relationship between the development of Ca2+ channel activity and neuronal survival is not understood. Over a period spanning 28 d in vitro, progressive increases in high voltage-activated whole-cell Ca2+ current and L-type Ca2+ channel activity were observed in cultured hippocampal neurons. On the basis of single-channel analyses, these increases seem to arise in part from a greater density of functionally available L-type Ca2+ channels. An increase in mRNA for the alpha1 subunit of L-type Ca2+ channels occurred over a similar time course, which suggests that a change in gene expression may underlie the increased channel density. Parallel studies showed that hippocampal neuronal survival over 28 d was inversely related to increasing Ca2+ current density. Chronic treatment of hippocampal neurons with the L-type Ca2+ channel antagonist nimodipine significantly enhanced survival. Together, these results suggest that age-dependent increases in the density of Ca2+ channels might contribute significantly to declining viability of hippocampal neurons. The results also are analogous to patterns seen in neurons of aged animals and therefore raise the possibility that long-term primary neuronal culture could serve as a model for some aspects of aging changes in hippocampal Ca2+ channel function. PMID- 9204945 TI - Nucleus reuniens thalami modulates activity in hippocampal field CA1 through excitatory and inhibitory mechanisms. AB - The nucleus reuniens thalami (RE) originates dense projections to CA1, forming asymmetrical synapses on spines (50%) and dendrites (50%). The hypothesis that RE input modulates transmission in CA1 through excitation of both pyramidal cells and interneurons was tested using electrophysiological methods in the anesthetized rat. The RE-CA1 afferents were selectively stimulated at their origin; evoked field potentials and unit activity were recorded in CA1. RE-evoked depth profiles showed a prominent negative deflection in the stratum lacunosum moleculare and a positive one in the stratum radiatum. The lacunosum-moleculare sink-radiatum source configuration is compatible with RE-elicited depolarization of apical dendrites of pyramidal cells. Despite a consistent and robust paired pulse facilitation of RE-evoked field potentials, population spikes in the stratum pyramidale were not detected at any tested condition. This indicates the inability of RE-CA1 input to discharge pyramidal cells. However, stimulation of RE-elicited spiking of extracellularly recorded units in strata oriens/alveus and distal radiatum, indicative of the activation of local interneurons. Thus, RE seems to modulate transmission in CA1 through a (subthreshold) depolarization of pyramidal cells and a suprathreshold excitation of putative inhibitory oriens/alveus and radiatum interneurons. RE-evoked monosynaptic or disynaptic field potentials were associated with stimulation of rostral or caudal RE, respectively. Anatomically, a projection from caudal to rostral RE was demonstrated that can account for the disynaptic RE-CA1 input. Because caudal RE receives input from the hippocampus via the subiculum, we propose the existence of a closed RE-hippocampal circuit that allows RE to modulate the activity in CA1, depending on hippocampal output. PMID- 9204946 TI - Lymphocyte activation in the pathogenesis of psoriasis. PMID- 9204947 TI - Selective amplification of T-cell receptor variable region species is demonstrable but not essential in early lesions of psoriasis vulgaris: analysis by anchored polymerase chain reaction and hypervariable region size spectratyping. AB - Several groups have investigated the role of T cells in the pathogenesis of psoriasis by determination of T-cell receptor (TCR) B-chain variable (V) region usage, both in chronic plaque (psoriasis vulgaris) and guttate forms, with various results. Because there are no data on TCR expression in early psoriasis vulgaris, when specific cellular immune events may be expected to be most pronounced, we have analyzed early lesions (less than 3 wk old) of ten patients, with highly reproducible results. We have developed a highly controlled anchored polymerase chain reaction (PCR) method in which TCR beta chain species are all amplified with the same primer pair and products are quantified by dot blot hybridization with BV family-specific oligonucleotide probes. Overexpression of certain TCR BV genes was observed in the majority of lesional biopsies, but in samples in which the expanded BV family formed more than 10% of total lesional BV (half of the samples analyzed), BV2 and BV6 predominated. The consistency of overexpression of these BV species between patients was much less than in previous studies of TCRBV usage in established chronic plaque psoriasis lesions. Complementarity-determining region 3 (CDR3) size spectratyping demonstrated evidence for selective clonal T cell accumulation in less than half of the lesional samples showing BV expansion. These results indicate that selective amplification of TCRBV species occurs in early psoriasis vulgaris but is not essential to the pathogenic process and may be more important in the maintenance or expansion of chronic lesions. PMID- 9204948 TI - The repertoire of T cell antigen receptor beta-chain variable regions associated with psoriasis vulgaris. AB - We investigated whether the pattern of T-cell receptors expressed by T cells in inflamed psoriatic skin differed substantially from the pattern seen in T cells from the peripheral blood. A bias or restriction in the repertoire of T-cell receptors found in the lesional skin of different patients might imply that specific subsets of T cells were causally associated with initiating or maintaining the lesions. By using a polymerase chain reaction-based assay of T cell receptor beta-chain variable region mRNA, we found that the patterns of beta chain mRNAs displayed in 14 samples of lesional skin or six samples of noninvolved skin were not significantly less diverse than the patterns found in matched peripheral blood samples. There was no evidence that the active lesions of multiple patients showed overexpression of T cells expressing one or a few T cell receptor forms. The pattern of T-cell receptors displayed in clinically normal skin from normal control individuals showed about the same diversity as normal blood. While these results may not exclude either classical antigen or superantigen-based T-cell activation mechanisms in active plaques, the absence of a simple pattern of Vbeta usage in different patients suggests than other aspects of T-cell biology including trafficking, proliferation, co-stimulation, or responses to cytokines must also be considered. PMID- 9204949 TI - Vitamin C abrogates the deleterious effects of UVB radiation on cutaneous immunity by a mechanism that does not depend on TNF-alpha. AB - Acute low-dose treatment of murine skin with ultra violet B (UVB) light impairs induction of contact hypersensitivity (CH) to dinitrofluorobenzene (DNFB) in certain inbred strains of mice (termed UVB-susceptible), but not in others (termed UVB-resistant), and promotes tolerance. These deleterious effects of ultraviolet radiation (UVR) are mediated in part by TNF-alpha, which is released from UVR-exposed epidermal and dermal cells. Because UVR damage to skin has also been ascribed in part to the generation of reactive oxygen intermediates (ROIs) such as superoxide anion (O2-), hydrogen peroxide (H2O2), hydroxyl radical (OH-), and singlet oxygen ((1)O2), we investigated whether vitamin C (ascorbic acid), which can nullify ROIs, prevents the deleterious effects of UVR on the cutaneous immune system. We found that epicutaneous application of vitamin C (10% L ascorbic acid solution) abrogated the deleterious effects of acute low-dose UVR on induction of CH and prevented the induction of tolerance. Vitamin C, however, did not reverse the effects of TNF-alpha on CH induction and tolerance. These results indicate that (i) ROIs generated intracutaneously by UVR contribute to the impaired ability of exposed skin to support the induction of CH and to promote the induction of tolerance and (ii) these effects are not dependent on TNF-alpha. PMID- 9204950 TI - Hapten-specific tolerance induced by acute, low-dose ultraviolet B radiation of skin is mediated via interleukin-10. AB - Because interleukin (IL)-10 is an immunoregulatory cytokine that is produced by keratinocytes exposed to UVB radiation (UVR), we determined whether IL-10 participates in either failed contact hypersensitivity (CH) induction or tolerance after acute, low-dose UVR. Murine recombinant IL-10 (200 ng) was injected intradermally on shaved abdominal skin. To assess the effects of IL-10 on CH induction, dinitrofluorobenzene (DNFB, 185 microg) was painted on the skin within 30 min after IL-10 was injected, and the mice were assayed 5 d later by ear challenge with dilute DNFB. To assess tolerance, DNFB (185 microg) was painted a second time on normal body-wall skin 14 d after DNFB was first painted on IL-10-injected skin; CH was then assayed on day 19. We found that mice that received DNFB on IL-10-injected skin developed CH comparable in intensity to that observed in PBS-injected controls. Thus, this dose of IL-10 did not prove to be deleterious to CH induction if hapten was painted on the injected site within 30 min. By contrast, mice that first experienced DNFB within 30 min, 1 d, or 3 d after IL-10 had been injected intracutaneously displayed hapten-specific tolerance. Moreover, intraperitoneally injected anti-IL-10 antibody prevented UVR and cis-urocanic acid-dependent tolerance; anti-IL-10 antibody had no effect on TNF-alpha-induced tolerance and failed to restore CH induction after UVR exposures. These data indicate that IL-10 is an important mediator of the tolerance induced when hapten is painted on the skin of animals exposed to acute, low-dose UVR. PMID- 9204951 TI - Pteridines in the control of pigmentation. AB - The influence of UVB irradiation on the metabolic pathway for the production of L tyrosine from L-phenylalanine in the human epidermis has been examined in 12 healthy volunteers with photo skin types I-VI (Fitzpatrick classification). This metabolic pathway involves the induction of GTP-cyclohydrolase 1 (GTP-CH-1), the rate-limiting enzyme for de novo synthesis of (6R)L-erythro-5,6,7,8 tetrahydrobiopterin (6-BH4). This essential cofactor controls the production of L tyrosine from L-phenylalanine via phenylalanine hydroxylase (PAH). The de novo synthesis of 6-BH4 depends on the induction of GTP-CH-1, e.g., by tumor necrosis factor-alpha (TNF alpha). Epidermal suction blister tissues were taken before (0 h) and after (24 and 72 h) UVB exposure with a standardized dosage [1 minimal erythema dose (MED)]. In all cases, there was a significant increase in TNF alpha release, GTP-CH-1 activity, total 6-biopterin level, and PAH activity, indicative of enhanced L-tyrosine production. The response of this metabolic cascade over baseline activities was pronounced in fair photo skin types (I-III) compared to dark skin (IV-VI). Taken together, our results suggest that UVB can control the direct supply of L-tyrosine in the epidermis, and this process may represent an important factor in de novo melanogenesis. PMID- 9204952 TI - Harlequin ichthyosis keratinocytes in lifted culture differentiate poorly by morphologic and biochemical criteria. AB - Harlequin ichthyosis (HI) is a severe congenital ichthyosis in which massively thickened stratum corneum with abnormal barrier function often results in death of affected newborns. Survivors evolve into a severe nonbullous ichthyosiform erythroderma. Previously we have ascertained three biochemical phenotypes of HI, based on abnormal profilaggrin and K6 and K16 expression in epidermis. Submerged cultures of HI keratinocytes differentiated abnormally, but the three phenotypes were indistinguishable in vitro. We hypothesized that differentiation in submerged culture was insufficient to reflect in vivo biochemical abnormalities or that dermal components might be necessary for expression. To test these hypotheses HI keratinocytes and fibroblasts (n = 3) were grown on collagen gels at the air-medium interface in a cross-over design with normal keratinocytes and fibroblasts. Epithelia derived from lifted cultures were studied by light microscopy and immunocytochemistry and extracted for western blot analysis. In contrast to our prediction, lifted cultures of HI keratinocytes formed a poorly differentiated epithelium, and normal keratinocytes formed an epidermal-like tissue with expression of K1 and expression and processing of profilaggrin to filaggrin. In addition, the presence of HI fibroblasts consistently altered differentiation of both HI and normal keratinocytes, resulting in less complete morphologic differentiation. The findings suggest that both epithelial and mesenchymal elements of the skin from HI are affected but that the primary abnormality lies in the keratinocytes. PMID- 9204953 TI - Response of SCC-12F, a human squamous cell carcinoma cell line, to complement attack. AB - We studied the response of a human squamous cell carcinoma cell line, SCC-12F, to human complement attack and found that the cells were completely resistant to complement lysis. In the absence of lysis, there was significant C3 deposition and C5b-9 deposition on the cells. Removal of the lipid-linked complement regulatory proteins CD59 and decay-accelerating factor (DAF) by treatment of the cells with phosphatidylinositol-specific phospholipase C (PIPLC) resulted in increased C3b and C5b-9 deposition on the cells and a slight increase in cell death. Treatment of the cells with complement caused them to release membrane vesicles containing the terminal complement proteins. In addition, complement induced SCC-12F to produce significant amounts of prostaglandin F2alpha (PGF2alpha). We conclude that CD59 and DAF are important in the resistance of SCC 12F to complement and that these cells produce membrane vesicles and PGF2alpha in response to complement attack. These responses, in the absence of cell death, may be important in the pathogenesis of inflammatory skin disease in which complement is deposited. PMID- 9204954 TI - Retinoic acid modulates the anti-proliferative effect of 1,25-dihydroxyvitamin D3 in cultured human epidermal keratinocytes. AB - Both 1,25-dihydroxyvitamin D3 (VD) and retinoids have potent effects on keratinocyte proliferation. Parallelism in their action as steroid hormones, which involves interaction of their receptors, and in their therapeutic efficacy for hyper-proliferative skin diseases provides a rationale to investigate their combined action on proliferation in pre-confluent human epidermal keratinocyte cultures. As shown by [3H]thymidine incorporation, all-trans retinoic acid (atRA) at subpharmacologic concentrations and 9-cis retinoic acid (9cRA) diminished the anti-proliferative effect of VD. Pre-incubation of the cells with the retinoids clearly enhanced this effect. Cell-cycle analysis revealed G1 arrest upon VD treatment that was attenuated by retinoic acid (RA). Moreover, Northern and Western blot analysis demonstrated that retinoic acid opposed VD-induced accumulation of transforming growth factor-beta1, p21WAF1, and p27KIP1. Finally, retinoic acid reduced VD-elicited hypophosphorylation of the retinoblastoma protein. AtRA at micromolar concentrations conversely potentiated most of the aforementioned VD-dependent actions. In addition, atRA and 9cRA (but not VD) caused a rapid, sustained reduction of RXR alpha protein. VD receptor protein was induced by VD regardless of the presence of RA. In conclusion, RA modulates VD dependent effects at different levels of keratinocyte proliferation. This could have implications for the use of combinations of both drugs for skin diseases. PMID- 9204955 TI - Liposome-associated interferon-alpha-2b functions as an anti-fibrogenic factor for human dermal fibroblasts. AB - This study was conducted to determine whether interferon-alpha-2b (IFN-alpha-2b) can be encapsulated in liposomes without compromising its anti-fibrogenic effects on human dermal fibroblasts. The rationale for this approach is that systemic administration of IFN-alpha-2b by injection for treatment of dermal fibrosis is uncomfortable, requires a large quantity of the cytokine, and cannot be easily used in children. Liposomes are potentially useful as vehicles for the topical delivery of drugs if they can be encapsulated without loss of biologic activity. Empty sonicated vesicles composed of dioleoyl-phosphatidylcholine:dioleoyl phosphatidylglycerol at a molar ratio of 7:3 were mixed with various concentrations of IFN-alpha-2b and then dried and rehydrated. An enzyme-linked immunosorbent assay (ELISA) was used to determine the efficiency of encapsulation and the stability of the preparation under experimental conditions. Greater than 80% of added IFN-alpha-2b became associated with the liposomes and remained encapsulated for up to 5 d at 4 degrees C. The rate of release increased markedly at 37 degrees C. Liposome-encapsulated IFN-alpha-2b (2000 units per ml) significantly reduced the proliferation of dermal fibroblasts (60 +/- 8.8 vs. 100 +/- 8, mean +/- SEM, p < 0.05, n = 8) and the levels of mRNA for type I (41.5 +/- 8.7% vs 100 +/- 18, p < 0.05, n = 4) and type III (68 +/- 8.4% vs 100 +/- 4.9%, p < 0.05, n = 3) procollagen, as analyzed on northern blots. This was consistent with the reduction found in collagen in conditioned medium from treated fibroblasts. In contrast, treatment increased levels of mRNA for collagenase (241 +/- 42% vs 100 +/- 3.4, p < 0.05, n = 3) and collagenase activity (289 +/- 5.8% vs 100 +/- 10.9%, p < 0.05, n = 9) in conditioned medium. This last effect was probably not due to a reduction in TIMP-1 (tissue inhibitor of metalloproteinase 1) because levels of mRNA for this inhibitor were not lower in treated cells. The efficacy of liposome-associated IFN-alpha-2b in vitro supports the concept of the topical use of this anti-fibrogenic agent for treatment of fibroproliferative disorders. PMID- 9204956 TI - Restoration of CDKN2A into melanoma cells induces morphologic changes and reduction in growth rate but not anchorage-independent growth reversal. AB - CDKN2A is a melanoma susceptibility gene that is mutated and/or deleted in familial and sporadic melanoma as well as in a range of other tumors. It encodes a cell cycle regulator, p16, whose function is to inhibit activity of cyclin dependent kinases 4 and 6. We set out to investigate the effect of reintroducing CDKN2A into MM96L, a melanoma cell line that does not express p16, by electroporation of wt CDKN2A cDNA. Our results show that transfection of the CDKN2A cDNA has a dramatic effect on cell morphology, inducing a more dendritic phenotype resembling that of adult melanocytes. This effect on cell morphology was not cell line specific because it was reproduced in another melanoma line (SK MEL-13), which has a homozygous deletion of CDKN2A. It was abolished by mutations that abrogate p16 function, as shown by transfection of a Pro81Leu p16 variant. Reintroduction of levels of p16 protein similar to those of cultured neonatal foreskin melanocytes decreased the growth rate of the transfected clones. Surprisingly, we did not see any effect on anchorage-independent growth or on the following melanoma markers tested by western blotting: p21/WAF1, tyrosinase related antigen 1, HMB45, and intermediate filament antigen. These data indicate that reintroduction into melanoma cells of wild type p16 at levels similar to cultured melanocytes can induce morphologic changes and suppress growth but is not sufficient to affect anchorage-independent growth. PMID- 9204957 TI - Detection of tyrosinase autoantibodies in patients with vitiligo using 35S labeled recombinant human tyrosinase in a radioimmunoassay. AB - Tyrosinase antibodies recently have been reported to occur frequently in patients with vitiligo. We describe the detection of tyrosinase antibodies in vitiligo patients using in vitro 35S-labeled human tyrosinase in a radioimmunoassay. Of 46 vitiligo sera examined in the assay, five (10.9%) were found to be positive for tyrosinase antibodies. In contrast, 20 control sera and sera from 10 patients with Hashimoto's thyroiditis were negative. Four of the sera positive in the radioimmunoassay were also positive in an ELISA using mushroom tyrosinase as antigen. Absorption studies indicated that pre-incubation with mushroom tyrosinase absorbed out the immunoreactivity of the positive sera in the radioimmunoassay, suggesting cross-reactivity, but this absorption was never complete, indicating that there are tyrosinase antibodies in human sera that do not react with the mushroom protein. There was no obvious association between the presence of tyrosinase antibodies and the age of the patients (range: 22-62 y), their duration of disease (range: 5-20 y), or the type of vitiligo (one segmental, one symmetrical/periorificial, three symmetrical), although the three patients with the highest antibody levels also had an associated autoimmune disorder (one with Graves' disease; two with autoimmune hypothyroidism). The results confirm that tyrosinase autoantibodies are present in the sera of vitiligo patients but at a low frequency. The technique described is sensitive and quantitative and allows the detection of conformational epitopes. It will be useful in longitudinal studies to determine the relation between the clinical features of vitiligo and tyrosinase antibody levels. PMID- 9204958 TI - A homozygous in-frame deletion in the collagenous domain of bullous pemphigoid antigen BP180 (type XVII collagen) causes generalized atrophic benign epidermolysis bullosa. AB - We report a missplicing event affecting the expression of bullous pemphigoid antigen BP180 (type XVII collagen) in a patient with generalized atrophic benign epidermolysis bullosa (GABEB). The segregation of the mutated allele in the family is consistent with the pathogenic role of the mutation. The homozygous mutation 2441-2A --> G disrupts a splice-site sequence in gene (BPAG2) for BP180 and results in an in-frame exon skipping within the collagenous ectodomain of the protein. The consequent deletion of 9 amino acids in the mutant BP180 is predicted to alter the structure of the homotrimer and is expected to exert a deleterious effect on stability of the protein that would account for the complete absence of immunoreactivity of the proband's skin to antibodies directed against BP180. These findings underscore the importance of structural integrity of the extracellular domain of BP180 for the stability of the protein. PMID- 9204959 TI - Sjogren-Larsson syndrome is caused by a common mutation in northern European and Swedish patients. AB - Sjogren-Larsson syndrome (SLS) is an autosomal recessive disorder characterized by congenital ichthyosis, mental retardation, and spastic diplegia or tetraplegia. Patients with SLS have deficient activity of fatty aldehyde dehydrogenase (FALDH), an enzyme involved in long-chain fatty alcohol oxidation. The cDNA encoding FALDH has recently been cloned and several different mutations have been found in SLS patients. We have now identified a point mutation (C943 - > T) in 7 of 19 kindreds of European descent, accounting for 24% of the SLS alleles. The C943T mutation was only found in patients of northern European ancestry from Sweden, the Netherlands, Germany, and Belgium. Haplotype analysis suggested that the patients carrying the C943T allele were distantly related. All four Swedish patients were homozygous for C943T, indicating that this mutation is probably the major cause of SLS in the inbred Swedish families. The mutation leads to the substitution of serine for the highly conserved proline 315 in the FALDH protein, and expression studies confirm that it destroys enzymatic activity. The mutation was readily detected with an MnlI restriction enzyme digestion test. The finding that C943T is a common SLS mutation in northern European and Swedish patients affords a rapid simple method for diagnosing SLS by screening patients for this mutation with DNA-based methods. PMID- 9204960 TI - trans-4-(Aminomethyl)cyclohexane carboxylic acid (T-AMCHA), an anti-fibrinolytic agent, accelerates barrier recovery and prevents the epidermal hyperplasia induced by epidermal injury in hairless mice and humans. AB - Because wounding the epidermis increases proteolytic activity and because disorders associated with barrier dysfunction have elevated protease activity, we studied the effect of protease inhibitors on the time course of barrier recovery and on the development of epidermal hyperplasia induced by repeated injury. After injuries to the epidermis produced by tape stripping, acetone treatment, or detergent (SDS) treatment that disrupt the barrier, a single application of 5% tranexamic acid [4-(aminomethyl)cyclohexane carboxylic acid, t-AMCHA], a well known anti-plasmin reagent, accelerated barrier recovery in both hairless mouse and human skin. In contrast, neither aminocaproic acid nor aminobutyric acid, inactive analogs of t-AMCHA, affected the time course of barrier recovery. Several trypsin-like serine protease inhibitors, e.g., leupeptin, TLCK, and PMSF, also accelerated barrier repair. In contrast other types of protease inhibitors, e.g., EDTA, pepstatin, N-ethylmaleimide, chymostatin, and TPCK, did not accelerate barrier recovery. We next evaluated the effects of daily topical application of t-AMCHA on epidermal hyperplasia, induced by repeated tape stripping or acetone treatment for 7 d. The degree of hyperplasia, quantified by the measurement of epidermal thickness, was reduced in both models by repeated applications of t-AMCHA. Finally, proteolytic activity in both human and mouse epidermis increased 1-2 h after epidermal injuries that disrupt the barrier. These results demonstrate that the inhibition of plasmin, a serine protease, accelerates barrier recovery and inhibits the epidermal hyperplasia induced by repeated barrier disruption, perhaps by decreasing the extent of attendant epidermal injury. PMID- 9204961 TI - Tazarotene-induced gene 2 (TIG2), a novel retinoid-responsive gene in skin. AB - Retinoids exert their biologic effects through two families of nuclear receptors, retinoic acid receptors (RARs) and retinoid X receptors (RXRs), which belong to the superfamily of steroid/thyroid hormone nuclear receptors. By using a subtraction hybridization approach, we have identified a cDNA sequence TIG2 (Tazarotene-induced gene 2), whose expression is up-regulated by the treatment of skin raft cultures by an RAR beta/gamma-selective anti-psoriatic synthetic retinoid tazarotene [AGN 190168/ethyl 6-[2-(4,4-dimethylthiochroman-6-yl) ethynyl] nicotinate]. The retinoid-mediated up-regulation in the expression of TIG2 was confirmed by Northern blot analysis. Upon sequencing, TIG2 was found to be a cDNA whose complete sequence was not in the GenBank and EMBL data bases. The TIG2 cDNA is 830 bp long and encodes a putative protein product of 164 amino acids. TIG2 is neither expressed nor induced by tazarotene in primary keratinocyte and fibroblast cultures. Thus, TIG2 is expressed and induced by tazarotene only when keratinocytes and fibroblasts form a tissue-like 3 dimensional structure. We further demonstrate that RAR-specific retinoids increase TIG2 mRNA levels. In contrast, neither RXR-specific retinoids nor 1,25 dihydroxyvitamin D3 increased TIG2 levels. Finally, we demonstrate that TIG2 is expressed at high levels in nonlesional psoriatic skin but at lower levels in the psoriatic lesion and that its expression is up-regulated in psoriatic lesions after topical application of tazarotene. PMID- 9204962 TI - Distinct populations of stromal cells express collagenase-3 (MMP-13) and collagenase-1 (MMP-1) in chronic ulcers but not in normally healing wounds. AB - Proteolysis is an intrinsic component of cutaneous wound repair and several matrix metalloproteinases have been shown to participate in various stages of this process. Therefore, we investigated the expression of a novel metalloproteinase, collagenase-3 (MMP-13), in normally healing cutaneous wounds and chronic venous ulcers. MMP-13 was expressed abundantly by fibroblasts deep in the chronic ulcer bed but was not detected in epidermis and all the acute wounds. The spatial expression of MMP-13 differed from that of collagenase-1 (MMP-1), which was prominently expressed by migrating keratinocytes and dermal cells located just beneath the wound surface. Northern blot hybridization did not reveal expression of MMP-13 by fibroblasts cultured on tissue culture plastic. In accordance with our in vivo findings, however, fibroblasts grown in a collagen gel produced MMP-13 mRNA abundantly. Our results suggest that MMP-13 can be induced in skin during wound repair after altered cell-matrix interactions. Although both MMP-1 and MMP-13 have the unique ability to degrade fibrillar collagens, their regulation and role during wound repair seem different. Collagenase-1 is critical for re-epithelialization, and MMP-13 most likely plays a role in the remodeling of collagenous matrix in chronic wounds. PMID- 9204963 TI - Human skin/SCID mouse chimeras as an in vivo model for human cutaneous mast cell hyperplasia. AB - Human skin xenografted to mice with severe combined immunodeficiency syndrome (SCID) was evaluated to determine the integrity and fate of human dermal mast cells. There was an approximately 3-fold increase in number of dermal mast cells by 3 mo after engraftment (p < 0.05). These cells were responsive to conventional mast cell secretagogues and were confirmed to be of human origin by ultrastructural characterization of granule substructure and by reactivity for the human mast cell proteinase, chymase. CD1a+ Langerhans cells, also bone marrow derived cells, failed to show evidence of concomitant hyperplasia, and increased mast cell number was not associated with alterations in number of dermal vascular profiles identified immunohistochemically for human CD31. RT-PCR analysis demonstrated human but not murine stem cell factor (SCF; also termed mast cell growth factor, c-kit ligand) mRNA in xenografts. Epidermal reactivity for stem cell factor protein shifted from a cytoplasmic pattern to an intercellular pattern by 3 mo after engraftment, suggesting a secretory phenotype, as previously documented for human cutaneous mastocytosis. The majority (>90%) of mast cells demonstrated membrane reactivity for human SCF at the time points of peak hyperplasia. These data establish SCID mouse recipients of human skin xenografts as a potential in vivo model for cutaneous mast cell hyperplasia. PMID- 9204964 TI - Protein expression of fibroblast growth factor receptor-1 in keratinocytes during wound healing in rat skin. AB - Fibroblast growth factors have been shown to play important roles in wound healing. To define their sites of action, we examined the expression of fibroblast growth factor receptor-1 (FGFR-1) during burn wound healing in rat skin by immunohistochemistry and western blot analysis. In cryostat sections of intact skin, little or no staining was observed. After a burn, however, staining for FGFR-1 was found in newly forming epidermis. The suprabasal layer of such epidermis, composed mostly of regenerating keratinocytes, was stained intensely, whereas keratinocytes in newly forming hair follicles were devoid of staining. Staining gradually decreased week by week after wound closure and was hardly visible 10 weeks after the burn, when the thickness of the epidermis had returned to the normal level. Staining was also found in small blood vessels and capillaries of granulation tissues of the dermis. Western blot analysis using the same antiserum was performed in the newly forming epidermis 10 d after the burn. A single band was detected with an apparent molecular weight of 120 kDa, corresponding to the short membrane-bound form of rat FGFR-1. Our study indicates that FGFR-1 is expressed during wound healing, mainly in regenerating epidermis and to some extent in blood vessels of the dermis. Fibroblast growth factors may affect the proliferation and differentiation of epidermal keratinocytes as well as angiogenesis in the dermis via the FGFR-1 expressed during wound healing. PMID- 9204965 TI - A novel mutation of a leucine residue in coil 1A of keratin 9 in epidermolytic palmoplantar keratoderma. AB - Keratin 9 mutation was examined in a Japanese kindred of epidermolytic palmoplantar keratoderma (EPPK), which is a dominantly inherited autosomal disorder of keratinization characterized by diffuse thickening of the palms and soles and by epidermolytic hyperkeratosis histologically. We report herein a novel mutation, a C --> G transversion at nucleotide position 541 that converts a leucine residue (CTC) to a valine (GTC) at codon 159. As in all other reported cases of keratin 9 mutation in EPPK, this mutation lies within the highly conserved coil 1A of the rod domain, which is considered to play a role in the correct alignment of the coiled-coil molecules. PMID- 9204967 TI - Microsatellite instability in primary and metastatic melanoma. PMID- 9204966 TI - A novel threonine --> proline mutation at the end of 2B rod domain in the keratin 2e chain in ichthyosis bullosa of Siemens. AB - We report a novel mutation in a case of ichthyosis bullosa of Siemens that results in a threonine --> proline substitution in a novel location, codon 485 in a highly conserved residue position of the IATYRKLLEGE consensus motif at the end of the 2B rod domain segment of the keratin 2e chain. The disease phenotype is consistent with the inappropriate substitution of a proline near the end of the rod domain, because it lies near the predicted molecular overlap region of coiled coil molecules, which is critical for the maintenance of the structural integrity of keratin intermediate filaments. PMID- 9204968 TI - Paul Langerhans sesquicentennial, July 23rd, 1997. PMID- 9204970 TI - A pilot study of all-trans retinoic acid in patients with Philadelphia chromosome positive chronic myelogenous leukemia. AB - Retinoids have significant antiproliferative effect against chronic myelogenous leukemia (CML) cells in vitro. We conducted a pilot study to investigate the clinical effect of all-trans retinoic acid (ATRA) in patients with CML. Thirteen patients with Philadelphia chromosome (Ph)-positive CML in late chronic phase (n=7), accelerated phase (n=5), or blastic phase (n=1) were treated. All had been previously treated and 12 (92%) had disease refractory to interferon-alpha therapy. They received ATRA 175 mg/m2 orally in two divided doses daily until disease progression. The median duration of therapy was 56 days (range 11 to 190). Only one patient in late chronic phase had a transient decrease in WBC counts; all other patients in late chronic phase showed no response to therapy. Four of the five patients in accelerated phase showed evidence of antileukemia effect manifested by a decrease in bone marrow and/or peripheral blood blasts, promyelocyte and/or basophil percentages. In all cases the response was transient. The patient in blastic phase had no evidence of antileukemic effect. The treatment was well tolerated with the major side-effects being headache, nausea, dry skin, and dry mucosal membranes. One patient required dose reductions due to toxicity. We conclude that in this population of patients with extensively treated, advanced stage, Ph-positive CML, ATRA alone is ineffective for long-term therapy. The antileukemia effect seen in some patients warrants further investigation of retinoids in other schedules and in combinations in patients with CML. PMID- 9204969 TI - Clinical and biological characteristics of acute promyelocytic leukemia in Taiwan: a high relapse rate in patients with high initial and peak white blood cell counts during all-trans retinoic acid treatment. AB - Acute promyelocytic leukemia (APL) patients treated with all-trans retinoic acid (ATRA) and chemotherapy have been shown to have better outcome than those treated with conventional chemotherapy alone. However, the biological characteristics of leukemic cells and their clinical implications in patients treated with ATRA have not been well established. In this study, the biological and clinical features of 30 APL patients were reported. The risk factors for relapse and for occurrence of retinoic acid (RA) syndrome, which might cause morbidity or mortality of patients after ATRA treatment, were also analyzed. All patients showed 15;17 translocation by cytogenetic and/or gene analysis. Patients in this study had higher white blood cell (WBC) counts and a higher incidence of additional abnormalities than those from other areas. The ratio of long (L) form to short (S) form PML-RAR alpha fusion transcript was 1.8:1, a value lower than that of Latino patients but higher than that of Italians. Leukemic cells from four patients showed coexpression of T cell-associated antigen CD2 which was highly correlated with S form fusion transcript. Nine (36%) of the 25 patients treated with ATRA developed RA syndrome; all but one were successfully controlled by corticosteroid. Complete remission (CR) rate was 84%. Patients with high WBC counts tended to develop RA syndrome and had increased risk of relapse. Isochromosome for the long arm of the derivative chromosome 17, ider(17q), as an additional chromosomal abnormality was also associated with poor outcome in this study. In conclusion, APL in this study showed some different biological characteristics compared with those reported in other areas. High WBC count was a risk factor for relapse and development of RA syndrome after ATRA treatment. The prognostic implication of the presence of ider(17q) needs further clarification. PMID- 9204971 TI - Expression of interferon regulatory factor (IRF) genes and response to interferon alpha in chronic myeloid leukaemia. AB - Interferon regulatory factors (IRF) 1 and 2 are DNA-binding proteins which control interferon (IFN) gene expression. IRF1 functions as an activator for IFN and IFN-inducible genes, whereas IRF2 represses the action of IRF1. Expression of the two regulatory genes is itself IFN-inducible. Because therapeutic responses of chronic myeloid leukaemia (CML) patients to IFN-alpha may be determined by intracellular levels of these two mutually antagonistic transcription factors, we have devised a competitive reverse-transcription polymerase chain reaction (RT PCR) assay which provides an estimate of the ratio of IRF1 to IRF2 expression in a given cell population. Analysis of peripheral blood leucocytes from 25 normal individuals showed that the IRF1:IRF2 ratio varied between 1.13 and 2.30 (mean +/ s.d. 1.49 +/- 0.33). Similar values were obtained for normal bone marrow specimens, with no significant difference between CD34+ and CD34- cells. In contrast, the IRF1:IRF2 ratio in leucocytes from CML patients showed a much wider variation (0.53-5.11). Eleven out of 130 patients in chronic phase had ratios above the normal range, whereas none of the 33 blast crisis samples had a ratio >2.5. Analysis of diagnostic specimens in 59 CML patients treated subsequently with IFN-alpha showed a high IRF1:IRF2 ratio of 5.11 in one of two patients who became complete responders; all the 53 patients with minimal or no cytogenetic response had ratios below 2.5. In a separate series of 97 CML patients studied after IFN-alpha therapy a highly significant correlation was found between the IRF1:IRF2 ratio and both the cytogenetic and the molecular response (ie low concentration of BCR-ABL transcripts) to treatment: 53 out of 115 prospectively analysed samples of good cytogenetic responders had ratios above 2.0, as opposed to only 13 out of 91 samples from poor responders (P < 0.0001; chi2 test). We conclude that a high ratio of IRF1/IRF2 expression may be associated with good cytogenetic and molecular response to IFN-alpha in CML. PMID- 9204972 TI - Apoptosis of human myeloid leukemia cells induced by an inhibitor of protein phosphatases (okadaic acid) is prevented by Bcl-2 and Bcl-X(L). AB - Okadaic acid, an inhibitor of serine/threonine protein phosphatases 1 and 2A has been shown to cause mitotic arrest and cell death of HL-60 and K562 cells. HL-60 cells express Bcl-2 and little or no Bcl-X(L), while K562 expresses Bcl-X(L) but not Bcl-2. Since phosphorylation/dephosphorylation reactions have been suggested to be involved in the regulation of Bcl-2, we planned to investigate whether the expression of Bcl-2, Bcl-X(L) and Bax, a protein that antagonizes the antiapoptotic function of Bcl-2, are regulated in myeloid leukemia cell lines (K562, KU812 and HL-60) treated with okadaic acid. Our results indicate that exposure of all three leukemic cell lines to nanomolar concentrations of okadaic acid causes a loss of viability by activation of an apoptotic process accompanied by a marked decrease in the expression of Bcl-2, Bcl-X(L) and Bax at both mRNA and protein level, but not of c-fos, vimentin and epsilon-globin, ruling out a non-specific effect of okadaic acid. Furthermore, constitutive expression of either Bcl-X(L) or Bcl-2 by gene transfer inhibited apoptosis triggered by okadaic acid in K562 cells. Thus, we suggest that protein phosphatases may be involved in maintaining the expression of bcl-2 family genes as part of the survival machinery of the cell. PMID- 9204973 TI - Bcl-x(L) is heterogenously expressed by acute myeloblastic leukaemia cells and is associated with autonomous growth in vitro and with P-glycoprotein expression. AB - The cells from approximately 70% patients with acute myeloblastic leukaemia exhibit autonomous growth characteristics in vitro, which have been associated with a poor response to therapy. We have previously shown that leukaemic cells with autonomous growth characteristics express high levels of bcl-2 and are relatively resistant to apoptosis. As bcl-x(L) is a bcl-2-related gene with anti apoptotic activity which also confers resistance to cytotoxic drugs we have studied its expression in AML in relation to cellular growth characteristics and to the expression of P-glycoprotein. Cells from 15 patients were studied. Immunoblotting demonstrated bands at 31 kDa corresponding to bcl-x(L) from the cells of all patients. Bcl-x(S) was not detected in any sample. Using standardised, quantitative flow cytometry, bcl-x(L) expression ranged from 0.25 x 10(5) to 4.24 x 10(5) bound FITC molecules, (median 1.35 x 10[5]). AML blasts with autonomous growth in vitro expressed more bcl-x(L) (median 1.76 x 10[5]) than those which did not (median 0.86 x 10(5), P=0.01). Quantitative bcl-x(L) expression strongly correlated with that of P-glycoprotein, also measured by quantitative flow cytometry using the MRK16 antibody (r=0.95, P < 0.001), but not with MRPr1. These results provide a further explanation for the poor prognosis associated with autonomous in vitro growth of AML blasts and illustrate that these cells may coexpress different modalities of resistance to cytotoxic drug therapy involving both anti-apoptotic pathways (bcl-x(L), bcl-2) and classic multidrug resistance (MDR1). The implication of these findings is that the use of agents to reverse MDR1 function in AML may be unsuccessful in the absence of strategies to reduce resistance to apoptosis. PMID- 9204974 TI - Sensitization of multidrug-resistant human leukemia cells with MDR1-targeted antisense and inhibition of drug-mediated MDR1 induction. AB - Increased expression of MDR1 is strongly implicated in the appearance of chemotherapeutic drug resistance in cancer, especially hematological malignancies. We therefore examined the potential of antisense oligonucleotides to inhibit MDR1 and restore sensitivity to drug-resistant human lymphoblastic cells (CCRF-CEM). Treatment with two different phosphorothioate-modified antisense sequences as well as a DNA-RNA hybrid sequence resulted in a 30 to 45% decrease in MDR1 expression as determined by staining with the monoclonal antibody MRK16 followed by flowcytometry (FCM) analysis. Further, inhibition of MDR1 expression persisted for 3 days after removal of oligonucleotides. Increased accumulation of rhodamine 123 and nearly a three-fold sensitization of cells to vincristine paralleled the reduction in staining with MRK16. Reversed or scrambled control sequences had no effect in any of the assays. During the course of these studies, we observed a 25 to 75% increase in MRK16 staining of cells treated with the chemotherapeutic agents daunorubicin and vincristine as well as by the resistance reversal agents verapamil and cyclosporin. Treatment of cells with antisense oligonucleotides prior to exposure to daunorubicin or cyclosporin reduced the increase in MRK16 staining. These results indicate that antisense targeted to MDR1 can sensitize drug-resistant leukemia cells and suggest that antisense treatment may prevent the emergence of MDR1-mediated drug resistance. PMID- 9204975 TI - Chromosomal breakpoints and changes in DNA copy number in refractory acute myeloid leukemia. AB - Comparative genomic hybridization (CGH) was used to detect changes in DNA copy number in 25 cases of refractory acute myeloid leukemia (AML). CGH detected changes in DNA copy number in nine AML (36%). Losses (82%) were more frequent than gains (18%). No high-level amplifications were detected in any of the cases. Losses involved minimal overlapping regions at 5q14q32, 7q31.2q32 and 12p12. The most frequent gain was detected at 8q. CGH gave normal results in all cases with a normal karyotype or a translocation as the sole aberration. The absence of high level DNA copy gains suggests that, in contrast to other malignancies, gene amplification is not an important mechanism for drug resistance in AML. In addition to 5q and 7q, known to be associated with disease refractoriness, 12p may be another region related to poor prognosis. PMID- 9204976 TI - Detection of 16 p deletions by FISH in patients with inv(16) or t(16;16) and acute myeloid leukemia (AML). AB - Deletions of sequences centromeric to the p-arm breakpoint have been described in a subset of patients with inv(16) and acute myeloid leukemia (AML) and reported to be associated with a relatively good prognosis. We have investigated 16 p deletions in a cohort of 15 patients with AML and inv(16) or t(16;16) and compared non-deletion and deletion patients in terms of clinical course. Patients were studied by fluorescence in situ hybridization (FISH) using cosmid zit14 as a probe to detect the presence of 16 p deletions in metaphase chromosomes of leukemic cells. While seven patients (47%) revealed no evidence of a deletion, five patients (33%) presented 16 p deletions, thus bringing further support to the relatively frequent occurrence of this event in inv(16) patients. Remarkably, two patients with inv(16) and one patient with t(16;16) showed a mosaicism of deletion and non-deletion metaphases suggesting the presence of two distinct leukemic cell populations. Results let us assume that 16 p deletions are not restricted to inv(16) and may occur subsequently to inv(16) or t(16;16). The presence of a 16 p deletion in a case of inv(16) associated with CBFB-MYH11 transcript type E indicates that deletions are not limited to CBFB-MYH11 transcript type A rearrangements. Survival of deletion patients was compared with that of non-deletion and mosaic ones. No significant differences were observed. The advantage of FISH for enumerative and quantitative assessment of submicroscopic rearrangements of clinical significance is further emphasized. PMID- 9204977 TI - DNA methylation patterns in the calcitonin gene region at first diagnosis and at relapse of acute lymphoblastic leukemia (ALL). AB - Aberrant DNA methylation can occur early in neoplastic transformation and may lead to the development of cancer. We describe the alterations of methylation patterns at the DNA sequence level which occurred in the 5' region of the calcitonin gene in lymphoblasts from 14 pediatric patients with acute lymphoblastic leukemia (ALL). The DNA methylation status of 25 CpG sites was determined by sequence analysis after bisulfite treatment of the DNA. This method showed that 13 out of 14 patients had increased numbers of methylated CpG sites in the calcitonin gene region at initial diagnosis when compared to control DNA from healthy individuals. The 5' region of the calcitonin gene appears to be methylated to a significantly higher degree in T lineage ALL compared to B lineage ALL (P < 0.01). Each of six ALL patients who were investigated at initial diagnosis and at relapse showed alterations in DNA methylation between the two stages. These six cases were also investigated by Southern blot analysis with methylcytosine-sensitive restriction enzymes and this method showed an increase in DNA methylation in only four of the six cases. The DNA sequencing method thus appears to be better suited to assess alterations of DNA methylation than Southern blot analysis. There are marked regional differences in the frequency of methylation of individual CpG sites and in the frequency of alterations between the two stages. Our results show that alterations in DNA methylation continue to occur from the initial stage to the relapse stage of ALL, suggesting that aberrant DNA methylation may play a role in tumor progression. PMID- 9204978 TI - Lack of ETV6 (TEL) gene rearrangements or p16INK4A/p15INK4B homozygous gene deletions in infant acute lymphoblastic leukemia. AB - Acute lymphoblastic leukemia (ALL) occurring in infants less than 1 year of age differs clinically and biologically from that observed in older children. Cytogenetically, 11q23 translocations are detected in approximately 50% of infant ALLs and fuse the 11q23 gene HRX with a variety of partner chromosomal loci. Overall, HRX rearrangements are detected molecularly in 70-80% of infant ALLs as compared to 5-7% of ALLs arising in older children. Two recently described molecular abnormalities in childhood ALL are ETV6 gene rearrangements and homozygous deletions of p16(INK4A) and/or p15(INK4B). Each of these abnormalities occurs in 15-20% of all childhood ALLs, and neither can be accurately identified by routine cytogenetic analyses. The incidence of these genetic abnormalities and their potential relationship to HRX gene status in infant ALL is unknown. Using Southern blot analyses, we determined ETV6 and p16(INK4A)/p15(INK4B) gene status in a cohort of infant ALLs. No ETV6 rearrangements or homozygous deletions (n=69) or homozygous p16(INK4A) and/or p15(INK4B) gene deletions (n=54) were detected in any of the infant ALLs. Therefore, ETV6 and p16(INK4A)/p15(INK4B) do not play a significant role in the pathogenesis of infant ALL, further emphasizing the distinctive biology of this subset of leukemias. PMID- 9204979 TI - Extensive analysis of the retinoblastoma gene in adult T cell leukemia/lymphoma (ATL). AB - The retinoblastoma susceptibility gene (Rb) plays a key role in regulating the cell cycle in association with cyclins and cyclin-dependent kinases (CDKs). Alteration of the Rb gene as well as CDK inhibitors (CDKIs) leads to deregulated cellular growth which promotes cancer formation. We examined the genomic configuration of the entire Rb gene in 40 primary adult T cell leukemias/lymphomas (ATL) and two ATL cell lines by Southern blotting and also by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) analyses. Homozygous loss of exon 1 was identified in one of 21 acute ATL, one of 15 chronic ATL, and none of four lymphomatous ATL samples. No point mutations were identified. Previously, we found that 10 of these same ATL samples had alterations of either p16(INK4A) (homozygous deletion) or p27(kip1) (homozygous deletion or point mutation). Although the numbers are very low, none of the samples with an aberrant Rb gene had an altered CDKI and vice versa, suggesting that both genes probably operate in a common pathway and alteration of either can provide these cells with a growth advantage. PMID- 9204980 TI - The human lysozyme gene undergoes stepwise demethylation during phagocyte maturation. AB - The lysozyme (LZM) gene provides a very useful model for studies of phagocyte maturation, because its protein synthesis is increased during myelopoiesis and thus most abundant in terminally differentiated and activated phagoyctes. LZM gene expression and DNA methylation were examined in various normal and transformed hematopoietic cells. Two shifts toward LZM gene demethylation coincided with upregulation of expression: activation of expression in myeloid precursor cells was associated with significant demethylation at a CpG dinucleotide within an Alu repeat in the 5' flanking region; high-level expression in different types of mature phagocytic cells was associated with complete demethylation at two additional, intragenic CpG sites contained in Alu sequences. The possibility that methylation changes occurring within the 5' region of the human lysozyme gene could be involved in the transcription of this gene is discussed, as well as a possible role for demethylation in the maintenance of distinct maturation stages during phagocyte development. PMID- 9204981 TI - Human A1, a Bcl-2-related gene, is induced in leukemic cells by cytokines as well as differentiating factors. AB - Based on previously published observations regarding the protective effects of interleukin-1 (IL-1) and tumor necrosis factor alpha (TNF alpha) against gamma radiation, alkylating agents and ultraviolet radiation, we hypothesized that the protection against such DNA damaging treatments can be the result of a 'stress' like response induced by these cytokines and mediated by early response cellular gene(s). By applying the mRNA differential display to RNA obtained from A549 lung carcinoma cell line that was incubated with 50 ng/ml IL-1 for 0, 1, 2, and 6 h, we identified several cDNA fragments that correspond to genes regulated by IL-1. The full length cDNA for one fragment was obtained using 5'RACE, cloned, sequenced, and found to be homologous to human A1, a Bcl-2-related gene. In this study, we report that the expression of human A1 is either absent or present at low levels in leukemic cells, while it is expressed in human bone marrow cells and abundant in peripheral blood progenitors. It is induced by IL-1 and TNF alpha in A549 lung carcinoma, bone marrow, and certain leukemic cells. A1 is also induced in leukemic cells during granulocytic or macrophage but not erythroid differentiation. In conclusion, this is the first demonstration that A1 is inducible by cytokines in human bone marrow and certain tumor cells as well as myeloid differentiation in leukemic cells. PMID- 9204982 TI - Rarity of dominant-negative mutations of the G-CSF receptor in patients with blast crisis of chronic myeloid leukemia or de novo acute leukemia. AB - It is likely that leukemia results, at least in part, from mutations that lead to a block in the normal process of differentiation. A defined region of the cytoplasmic domain of the granulocyte colony-stimulating factor receptor (G-CSF R) transmits signals for maturation or differentiation of myeloid progenitor cells. Mutations in this region have been found in some patients with severe congenital neutropenia (SCN) who subsequently evolved to acute myeloid leukemia (AML). To determine if mutations of the G-CSF-R are more widespread in hematological malignancies, we have investigated a total of 47 patients, including 29 patients with blast crisis of chronic myeloid leukemia (CML-BC) and 18 patients with de novo acute leukemia as well as 19 normal controls, by RT-PCR and SSCP analysis. Two point mutations were found in a single individual with secondary AML (FAB type M1). The first was heterozygous and is predicted to replace the normal glutamine at position 718 with a stop codon, leading to a truncated protein. An identical mutation has been described previously and shown to act in a dominant negative manner. The second mutation was homozygous and would substitute a lysine for the normal glutamic acid at position 785. No mutations were found in any other patient or control samples. We conclude that mutations in the cytoplasmic domain of the G-CSF-R are infrequent in CML-BC or acute leukemia but may contribute to malignant transformation in some cases. PMID- 9204983 TI - Sezary cell leukaemia: a distinct T cell disorder or a variant form of T prolymphocytic leukaemia? AB - We report the clinical, ultrastructural, immunophenotypic and virological features of nine cases of a rare type of mature T cell disorder formerly designated Sezary cell leukaemia. All patients presented with lymphocytosis ranging from 12.7 to 133 x 10(9)/l, bone marrow infiltration, splenomegaly and lymphadenopathy. Skin involvement was absent at presentation but developed as a terminal event in two patients, one of whom showed a pattern of dermal infiltration different from that characteristic of Sezary syndrome. Cells from eight cases bore a mature T cell phenotype and electronmicroscopy revealed lymphocytes with cerebriform nuclei resembling Sezary cells. All cases except one were HTLV-I negative. Patients were treated with various chemotherapy regimens but with poor outcome, the median survival being 13 months. Laboratory and clinical data suggest great similarity between Sezary cell leukaemia and T prolymphocytic leukaemia (T-PLL), namely coexpression of CD4 and CD8 (3/9 cases), identical chromosomal abnormalities in the three cases studied (isochromosome 8q plus inversion 14 or t(X;14)(q28;q11)) and a remarkable sensitivity to CAMPATH-1H (complete remission of 21 months' duration in one patient), suggesting that this entity could be considered a variant form of T-PLL. The alternative diagnosis of adult T cell leukaemia/lymphoma could not be excluded in one patient in whom positive HTLV-I serology was documented. PMID- 9204984 TI - Differentiation of t(5;17) variant acute promyelocytic leukemic blasts by all trans retinoic acid. AB - All-trans retinoic acid (ATRA) induces differentiation of acute promyelocytic leukemic (APL) blasts from patients with t(15;17) APL. However, blasts from patients with the t(11;17) variant do not differentiate in response to ATRA. Our group has identified a variant of APL characterized by t(5;17) and expression of the NPM-RAR fusion gene product. From case reports it has been difficult to establish whether ATRA induces clinical responses in patients with this variant. In order to determine whether t(5;17) blasts differentiate with ATRA, we harvested mononuclear bone marrow cells from a patient with t(5;17) APL at time of relapse and cultured them in medium containing ATRA. Morphologic analysis of cytospins after 7 days of culture revealed that 60% of cells in the ATRA-treated culture had differentiated into mature neutrophilic forms, as opposed to less than 1% in the control culture. Seventy-three percent of cells acquired NBT positivity after exposure to ATRA, compared with 1% in the control culture. These results indicate that t(5;17) blasts retain the ability to terminally differentiate in response to retinoic acid. PMID- 9204985 TI - Interaction of vitamin D derivatives and granulocyte-macrophage colony stimulating factor in leukaemic cell differentiation. AB - The ability of the physiologically active form of vitamin D, 1,25 dihydroxyvitamin D3 (1,25(OH)2D3) and two novel vitamin D analogues, EB1089 and KH1060 to induce the differentiation of the U937 and HL-60 leukaemic cell lines was evaluated, alone or in combination with granulocyte-macrophage colony stimulating factor (GM-CSF). Studies revealed that following 96 h treatment, the vitamin D derivatives inhibited the proliferation, and induced the differentiation of U937 and HL-60 cells in a dose-dependent manner, as determined by cell counts and nitroblue tetrazolium (NBT) reduction assays, respectively. EB1089 and KH1060 were found to be more effective than 1,25(OH)2D3 in exhibiting their antiproliferative and differentiative effects. In contrast, induction of leukaemic cell differentiation with 1 ng/ml GM-CSF after 96 h was less effective when compared with the vitamin D derivatives used individually. Fluorescence activated cell scanning (FACS) analyses indicated that the vitamin D derivatives readily induced the expression of the monocyte-associated cell surface antigen, CD14, and also the beta2-integrins, CD11b and CD18 in both cell lines after 48 h and 96 h treatment. The ability of EB1089 and KH1060 to induce these antigens was achieved with greater efficacy relative to the native hormone. When U937 and HL 60 cell cultures were cotreated for 48 h with the vitamin D compounds and GM-CSF and analysed by FACS, enhanced effects on CD14 and CD11b induction were observed compared to those of the compounds alone. These co-operative effects may occur as a consequence of molecular events which involve the transcription by vitamin D receptors (VDR) of genes required for the responsiveness of immature cells to factors such as GM-CSF, and place these and other related vitamin D analogues as potential therapeutic agents in the treatment of leukaemia. PMID- 9204986 TI - Recurring proviral integration suggests a role for proto-oncogene activation in thymomas induced with Mo-MuLV-rescued BCR/ABL virus. AB - Intrathymic injection of Moloney murine leukemia virus (Mo-MuLV)-pseudotyped bcr abl retrovirus (bcr-abl/M) causes thymic lymphoma but only after a prolonged latent period similar to that seen after intrathymic injection of Mo-MuLV alone. Since thymomas induced by Mo-MuLV show recurring proviral integration near certain cellular proto-oncogenes, it was reasoned that if the pathogenesis of bcr abl/M thymomas is affected by viral integration, then it may be possible to detect proviral insertion near common Mo-MuLV integration sites in bcr-abl induced thymomas. A panel of thymomas induced by intrathymic injection of Mo MuLV, Abelson murine leukemia virus (A-MuLV), or the bcr-abl/M virus was analyzed for proviral integration near c-myc, N-myc, Pim-1, and Mlvi-1 loci that are frequently occupied by provirus in Mo-MuLV-induced T cell lymphomas, and for integration near Ahi-1 that is often occupied in A-MuLV/M-induced pre-B cell lymphoma. As expected, thymomas induced with Mo-MuLV showed frequent rearrangements in these loci while thymomas induced with A-MuLV/M (which does not require Mo-MuLV) did not. The bcr-abl/M-induced tumors also showed recurring proviral integration near c-myc, Pim-1 and Mlvi-1, albeit at a lower frequency than seen in the Mo-MuLV tumors. Unexpectedly, four independent thymomas that were clearly of T cell origin demonstrated proviral integration within the Ahi-1 region which was previously thought to only occur in A-MuLV/M induced pre-B cell lymphoma. These observations suggest that recurring proviral insertion in c-myc, Pim-1, Mlvi-1, and Ahi-1 may provide a selective advantage for bcr-abl/M transformed T lymphoid cells. This model may provide a tool for identifying cellular genes that can cooperate with bcr-abl in lymphoid transformation. PMID- 9204987 TI - Involvement of peripheral blood cells in multiple myeloma: chromosome changes are the rule within circulating plasma cells but not within B lymphocytes. AB - The mononuclear cells in the peripheral blood are implicated in the myeloma process especially with the presence of peripheral blood plasma cells (PBPC) and clonal B lymphocytes found using phenotypic or gene rearrangement techniques. The purpose of this study was to look for aneuploidy in the two main B cell components of the peripheral blood: PBPC and CD20-positive B lymphocytes. Conventional cytogenetics (CC) or DNA content analysis and fluorescence in situ hybridization (FISH) with centromeric probes were performed on bone marrow plasma cells (BMPC) of 21 patients with multiple myeloma and peripheral blood cells were studied as follows: immunostaining to look for PBPC and to assess their number, image analysis cytometry for the determination of their DNA content, and FISH chromosomes analysis. FISH was performed using probes against the chromsomes that were lost or gained in BMPC and was coupled with immunostaining of the relevant light chain or CD20 antigen to study PBPC or B lymphocytes, respectively. Monotypic PBPC were found in 16 patients. Their DNA content was the same or nearly the same as for BMPC and they exhibited the same monosomies or trisomies as those found within their BM counterpart. By contrast, DNA content of mononuclear cells other than PBPC was within normal ranges, and in 13 of 15 patients CD20-positive B lymphocytes failed to show chromosomal changes by FISH analysis. In two patients however, a few CD20+ cells with lymphoid morphology exhibited chromosome changes, hypothesizing that a few cytogenetically abnormal B cells without plasmocytic morphology may circulate. From these data, we conclude that PBPC share the same genetic abnormalities as BMPC and thus belong to the malignant clone, whereas most peripheral blood B lymphocytes are unrelated to the tumor clone. PMID- 9204988 TI - A new human cell line with pre-B cell phenotype and t(5;12). AB - A new cell line (LR10.6) with pre-B cell phenotype has been established from bone marrow cells obtained from a child with B lineage acute lymphoblastic leukemia in complete clinical remission. The line expresses nuclear TdT enzyme, cytoplasmic Ig lambda-chain and membrane mu-chain and other B but no T or myeloid markers. The cells also show activation antigens CD69 and CD71, adhesion molecules CD54, CD50 and CD56 and the tyrosine kinase receptor CD117. No expression of multidrug resistance phenotype MDR-1 is observed on these cells which nevertheless express the transcriptional factor p53 protein in a mutant form. Cytogenetic study shows a translocation t(5;12)(q31;p13) involving breakpoints which contain the growth factor interleukin 3 gene (5q31) and the recently identified TEL/ETV6 gene (12p13). Activation of the cells with phorbol-12 myristate 13-acetate (PMA) up regulates the expression of the CD69 activation antigen and down-regulates the CD117 molecule. In addition, PMA fails to induce the CD20 B cell antigen. PMID- 9204989 TI - Increased rejection of murine allogeneic bone marrow in presensitized recipients. AB - The role of presensitizing murine recipients with donor spleen cells prior to T cell-depleted or -repleted H-2 compatible allogeneic bone marrow transplantation (BMT) was investigated at two different doses of total body irradiation (TBI). Recipients that were presensitized with 2 x 10(7) irradiated donor spleen cells at 1 week before a sublethal dose of 6 Gy TBI and BMT showed no evidence of donor blood chimerism while unsensitized recipients showed about 80% donor engraftment as determined by blood Gpi phenotyping. After raising the TBI dose to 9.5 Gy an increase in mortality from marrow failure was observed in presensitized animals. No significant engraftment-promoting effect of up to 2 x 10(6) T cells (20% of total marrow dose) was seen either in presensitized or unsensitized mice. It can be concluded that presensitized recipients are more susceptible to acute marrow rejection and that T cells added to the bone marrow did not influence the level of donor engraftment in these recipients. PMID- 9204990 TI - Facilitated engraftment of human hematopoietic cells in severe combined immunodeficient mice following a single injection of Cl2MDP liposomes. AB - Transplantation of normal and malignant human hematopoietic cells into severe combined immunodeficient (SCID) mice allows for evaluation of long-term growth abilities of these cells and provides a preclinical model for therapeutic interventions. However, large numbers of cells are required for successful engraftment in preirradiated mice due to residual graft resistance, that may be mediated by cells from the mononuclear phagocytic system. Intravenous (i.v.) injection of liposomes containing dichloromethylene diphosphonate (Cl2MDP) may eliminate mouse macrophages in spleen and liver. In this study outgrowth of acute myeloid leukemia (AML) cells and umbilical cord blood (UCB) cells in SCID mice conditioned with a single i.v. injection of Cl2MDP liposomes in addition to sublethal total body irradiation (TBI) was compared to outgrowth of these cells in SCID mice that had received TBI alone. A two- to 10-fold increase in outgrowth of AML cells was observed in four cases of AML. Administration of 10(7) UCB cells reproducibly engrafted SCID mice that had been conditioned with Cl2MDP liposomes and TBI, whereas human cells were not detected in mice conditioned with TBI alone. As few as 2 x 10(4) purified CD34+ UCB cells engrafted in all mice treated with Cl2MDP liposomes. In SCID mice treated with macrophage depletion unexpected graft failures were not observed. Histological examination of the spleen showed that TBI and Cl2MDP liposomes i.v. resulted in a transient elimination of all macrophage subsets in the spleen, whereas TBI had a minor effect. Cl2MDP liposomes were easy to use and their application was not associated with appreciable side-effects. Cl2MDP liposome pretreatment in combination with TBI allows for reproducible outgrowth of high numbers of human hematopoietic cells in SCID mice. PMID- 9204991 TI - Automated high resolution PCR fragment analysis for identification of clonally rearranged immunoglobulin heavy chain genes. AB - The development of rapid polymerase chain reaction (PCR) protocols for amplification of rearranged heavy chain immunoglobulin (IgH) gene sequences has facilitated the identification of clonal IgH rearrangements in non-Hodgkin's lymphomas (NHL) and leukemias of B lineage. In the present report we have explored the recently described improved strategy for assessment of clonality of rearranged immunoglobulin heavy chain (IgH) genes in more detail in a series of 101 B cell malignancies and 50 polyclonal controls. The assay is based on an IgH PCR with an automated fluorescence-based strategy for PCR detection of IgH gene rearrangements. Third complementarity determining region (IgH-CDR3) sequences were amplified using fluorescent dye labeled consensus primers homologous to the corresponding variable (V[H]) and joining (J[H]) gene segments in combination with a thermostable proofreading DNA polymerase. PCR products were size separated on a high resolution polyacrylamide gel and analyzed for clonality by exact size determination and fluorescence quantification in an automated DNA sequencer. PCR findings obtained with the optimized IgH-CDR3-PCR assay showed an overall monoclonality detection rate of 97% (97 of 101 cases with B cell neoplasms). The specificity was 100% as determined by analysis of 50 controls, all of which gave polyclonal PCR results. We found a high rate of monoclonal IgH-CDR3-PCR results not only in the leukemias and diffuse lymphoma but also in the group of follicular lymphoma, where a high rate of false negative results is frequently reported in the literature. In summary, we identified monoclonal IgH-CDR3 junctions in 55 out of 59 cases (93%) with B cell lymphoma and in 42 of 42 (100%) cases with leukemia, immunocytoma and multiple myeloma. The results demonstrate that automated fluorescence detection of IgH-CDR3-PCR products is an ideal tool for detection of clonal and polyclonal lymphoid B cells. In combination with allele-specific primers the procedure may improve current experimental approaches to detect occult malginant B cells during initial staging and follow-up of NHL and ALL patients. PMID- 9204992 TI - Phenotypic and genotypic analyses of multidrug resistance (MDR) in clinical hospital practice. PMID- 9204993 TI - Multidrug resistance in acute leukemia: a comparison of different diagnostic methods. AB - Accurate measurement of P-glycoprotein (P-170) expression in clinical samples still remains a controversial issue. In this study tumor cell P-170 expression was assessed in 29 patients suffering from acute leukemia (17 acute myeloid leukemia (AML) and 12 acute lymphoblastic leukemia (ALL)) using three different techniques: flow cytometry measuring rhodamine 123 (Rh123) efflux (functional level), immunocytochemistry (protein level) and RT-PCR (mRNA level). Rh123 efflux was detectable in 10/29 (34%) of all cases, in 9/17 (53%) of AML and in 1/12 (8%) of ALL samples. In AML patients a significant association of CD34 expression and P-170 activity was observed (P < 0.02). All AML patients with the FAB subtype M5 were Rh123 negative (P < 0.007). Cytospin preparations were analyzed for staining with monoclonal antibodies JSB1 and MM4.17. Eight of 16 (50%) AML and 0/9 (0%) ALL cases expressed the multidrug resistance (MDR) protein assessed by JSB1. With MM4.17 87% of AML and 50% of ALL patients were scored positive. Agreement between both antibodies was found in only 13/23 (57%) samples. Extracted RNA from 12 patients was analyzed by RT-PCR to evaluate the expression of MDR1 and multidrug resistance-associated protein (MRP) mRNA. An increased level of MDR1 mRNA was detectable in 4/7 AML and 0/5 ALL cases. MRP expression was found in 3/7 AML and 0/5 ALL patients. Comparison of Rh123 assay and immunocytochemistry revealed a very good correlation when using MoAb JSB1 (P < 0.004) but not with MM4.17 (not significant (NS)). JSB1 also showed a much better association with the PCR results (P < 0.05) than MM4.17 (NS). Finally, we compared the results of the functional Rh123 assay and RT-PCR and observed a high correlation for Rh123/MDR1 (r = 0.819, P < 0.001) but low for Rh123/MRP (r = 0.562, NS). We conclude that measurement of Rh123 efflux and immunocytochemical staining of cytospin preparations with JSB1 allows the accurate monitoring of P-170 expression in acute leukemia. The simplicity of these two MDR assays suggests their use for routine MDR screening. PMID- 9204994 TI - Immunocytochemical detection of the multidrug resistance-associated protein and P glycoprotein in acute myeloid leukemia: impact of antibodies, sample source and disease status. AB - Immunocytochemical detection of the expression of the MRP gene and the MDR1 gene in clinical specimens might be affected by several factors. Thus, we studied the impact of monoclonal antibodies, sample source (peripheral blood vs bone marrow) and disease status on the expression of multidrug resistance-associated protein (MRP) as well as P-glycoprotein (P-gp) in leukemic cells of patients with acute myeloid leukemia (AML). MRP expression was determined by means of anti-MRP antibodies (QCRL-1, QCRL-3, QCRL-1/QCRL-3 or MRPr1). In the case of P-gp, monoclonal antibodies C219 and MRK16 were used. High MRP expression ranged from 5 to 35% and high P-gp expression from 5 to 14% of the specimens. A fair correlation between results obtained with QCRL-1/QCRL-3 and those obtained with MRPr1, as well as a moderate correlation between C219 and MRK16, were seen. MRP and P-gp expression of peripheral blood blasts were similar to those of bone marrow blasts in the majority of cases. The degrees of MRP expression at the time of diagnosis were also similar to the degrees of expression at relapse, albeit an analysis of sequential MRP expression in 13 patients indicated an increase of expression at relapse in six patients as compared to the time of diagnosis. PMID- 9204995 TI - Optimal immunocytochemical and flow cytometric detection of P-gp, MRP and LRP in childhood acute lymphoblastic leukemia. AB - The clinical relevance of multidrug resistance (MDR)-related proteins in childhood acute lymphoblastic leukemia (ALL) is largely unknown. The diversity of techniques, fixation methods, storage of cells (fresh or cryopreserved) etc, may contribute to discrepancies observed between several studies. We therefore optimized the detection of P-glycoprotein (P-gp), MDR-associated protein (MRP) and lung resistance-related protein (LRP) by immunocytochemistry and flow cytometry in childhood ALL cells. Thirteen fixation methods were compared using six antibodies in both immunocytochemistry and flow cytometry. The optimal fixation for P-gp (C219, MRK16), MRP (MRPr1) and LRP (LRP56) was a mixture of 2% (v/v) formaldehyde solution and acetone incubated for only 10 s at room temperature (FAc). For MRP recognized by MRPm6, the optimal fixation condition was acetone for 5 min at room temperature in immunocytochemistry, and methanol for 15 min at -20 degrees C in flow cytometry. P-gp staining by 4E3 was strongly antibody batch-dependent; on cytospins FAc fixation was optimal, but inconclusive data were obtained by flow cytometry. The optimized fixation conditions on fresh samples revealed a day-to-day variation in staining (both increasing and decreasing) in one third of the immunocytochemical tests. In flow cytometry the day-to-day variation in the fluorescence index was -1 +/- 22%. In both techniques, staining was comparable between fresh and cryopreserved cells. We recommend the use of the above mentioned fixation methods in order to study the clinical relevance of P-gp, MRP and LRP in childhood ALL. PMID- 9204996 TI - Multicentric evaluation of the MDR phenotype in leukemia. French Network of the Drug Resistance Intergroup, and Drug Resistance Network of Assistance Publique Hopitaux de Paris. AB - The wide discrepancies in the frequency of 'positive' samples for multidrug resistance (MDR) phenotype within the same type of tumor observed in the literature justified the need for the definition of consensus recommendations. To define standard techniques of MDR phenotype measurement, we ran a large multicentric evaluation of the different methods available. Thirty-six French centers participated in the study, and 742 samples of 2-10 x 10(6) viable cells were sent by overnight express mail between December 1993 and February 1996. The same batches of MRK16, 4E3 and UIC2 were used. Nineteen samples of leukemia (12 AML, 1 ALL, 6 lymphoproliferative syndromes) and six leukemic cell lines with different levels of MDR expression were tested. Five meetings reached agreement concerning the guidelines for each technique, except immunocytochemistry. The 19 fresh samples were tested by each center using one to four techniques among cytofluorometry, immunocytochemistry, functional tests and RT-PCR. Five samples were diagnosed as 'negative' according to local criteria, with few discordant results (0 to 16% of 'positive' results). For all the 14 remaining samples, large discrepancies were observed from center to center, and from one technique to another. No correlations could be found between techniques. Flow cytometric analysis of cells already exposed to MRK16 or control IgG2A, fixed in paraformaldehyde and sent to centers did not reduce the discrepancies between centers in two of the four samples with moderate expression, emphasizing the role of histogram interpretation. The use of alternative monoclonal antibodies (4E3 and UIC2) did not reduce the discrepancies observed. In a second step, the K562 parental cell line, a low resistant subline (K562/HHT100, x7 resistance index to DNR) and a high resistant subline (K562/HHT300, x125 resistance index to DNR) were sent blindly three times, with an increasing level of recommendations for flow cytometry. Dramatic improvements were observed in cytometric results when the result was expressed as the ratio of arithmetic mean of fluorescence of antibody (10 microg of MRK16)/arithmetic mean of fluorescence of control (10 microg IgG2A): the proportion of expected results increased from 61 to 100% for K562, and from 37 to 85% for K562/HHT100. For uptake and drug efflux measurements, the use of 1 h uptake of 0.1 microM of rhodamine, followed by 1 h efflux +/-10 microM of verapamil, permitted an increased reproducibility of the technique from 71 to 100% for K562 and K562/HHT100. Whatever the technique used, concordant results were obtained for K562/HHT300. The immunocytochemistry, using several antibodies (MRK16, JSB1 and C219) gave many non-interpretable results (44%), due to a frequent high background and discordant results between antibodies in the same centers, and discordant conclusions between centers. The group does not recommend this technique for circulating tumoral cells. PMID- 9204998 TI - Methods to detect P-glycoprotein and implications for other drug resistance associated proteins. AB - The problem of tumor cell drug resistance remains a barrier to the successful treatment of many neoplastic diseases. Problems of tumor cell heterogeneity and expression of multiple mechanisms of drug resistance complicate treatment strategies. Indeed, even that form of resistance to natural product anticancer agents, multidrug resistance (MDR), can have multiple mechanisms. Compounding these problems is the use of different methodologies and different reagents to assess expression of the most widely studied form of MDR, that due to increased expression of the MDR1 gene and its product, P-glycoprotein (Pgp). In this paper, we discuss problems associated with assay variability and accurate measurement of markers of drug resistance, and summarize consensus findings of the St Jude Workshop on methods to detect Pgp in tumors. PMID- 9204997 TI - French multicentric evaluation of mdr1 gene expression by RT-PCR in leukemia and solid tumours. Standardization of RT-PCR and preliminary comparisons between RT PCR and immunohistochemistry in solid tumours. French Network of the Drug Resistance Intergroup, and Drug Resistance Network of Assistance Publique Hopitaux de Paris. AB - Since there is no consensus on the techniques for multidrug resistance (MDR) phenotype evaluation, many discrepancies concerning the importance and frequency of mdr1 gene expression in leukemias and solid tumors are observed in the literature. In order to establish an inter-laboratory consensus in France, a multicenter study was carried out to propose further guidelines for MDR phenotype evaluation. The techniques used by the 38 laboratories participating in the trial were: immunodetection (immunohisto and/or cytochemistry, flow cytometry), functional tests, reverse transcription-polymerase chain reaction (RT-PCR) or Northern blot. We present the results obtained by 19 laboratories concerning the measurement of mdr1 gene expression assessed by RT-PCR or Northern blot in: (1)19 samples of tumor cells obtained from leukemic patients; (2) six solid tumor samples obtained at surgery; (3) eight cell lines exhibiting variable levels of resistance, and; (4)10 preparations of RNA and of cDNA obtained from solid tumors. Standardization of the RT-PCR technique and preliminary results comparing RT-PCR with immunohistochemistry in solid tumors are also reported. PMID- 9204999 TI - Theoretical and practical considerations for the measurement of P-glycoprotein function in acute myeloid leukemia. AB - This paper summarizes experimental data and theoretical considerations, that are important for the measurement of P-glycoprotein (Pgp) function in acute myeloid leukemia (AML). The data are presented in subdivisions based on the techniques used, which will facilitate finding specific information. Based on our extensive experience with Pgp analysis, which includes radioactive assays, flow cytometry and fluorescence microscopy, we recommend a flow cytometry-based assay, that measures the effect of 2 microM PSC 833 on rhodamine 123 (R123) accumulation as the most practical and sensitive functional Pgp test. In combination with the flow cytometric measurement of Pgp using an antibody against an extracellular epitope (eg MRK16), this offers a sensitive and reproducible method for Pgp detection in AML, which is also rapid and practical. Furthermore, an R123 accumulation assay is specific for Pgp, because R123 is transported much less efficiently by the multidrug resistance protein (MRP) than by Pgp. Another probe of similar sensitivity and specificity is 3,3'-diethyloxacarbocyanine iodide. Alternatively, especially for the analysis of small numbers of cells (for example sorted subpopulations of leukemic cells), convenient and sensitive procedures are being developed by using DNA-binding Pgp substrates which remain fixed in the nuclei of the cells upon formaldehyde exposure for quantitative fluorescence laser scanning microscopy with image analysis. Less experimental data have been published to establish the optimal conditions for dual parameter flow cytometry (Pgp function, in eg Pgp+ or CD34+ cells). However, laboratories with flow cytometry experience will be able to implement this useful option to analyze subpopulations of cells. PMID- 9205000 TI - Monoclonal antibodies specific for P-glycoprotein. AB - Multidrug resistance (MDR) is one of the major obstacles to successful cancer chemotherapy. Since P-glycoprotein (P-gp) encoded by the MDR-1 gene plays a key role in MDR, many P-gp-specific monoclonal antibodies (MAbs) have been generated for characterization and analysis of P-gp. Among those antibodies, MRK16 has been widely used not only for elucidation of the mechanisms of P-gp-mediated MDR but also for diagnostic and therapeutic studies. Two types of magnetic cell sorting assays, termed MRK16-MACS and MRK16-MACS-FACS, have been established by us and may offer a useful tool to quantitate low levels of P-gp expression. This article describes the characteristics of the antibodies against P-gp and discuss the diagnostic implications of the antibodies. PMID- 9205001 TI - Flow cytometric analysis of P-glycoprotein function using rhodamine 123. AB - The MDR1 gene product, P-glycoprotein (P-gp), works as a transmembrane efflux pump for several cytotoxic products, representing a major cause for cancer treatment failure. Rhodamine 123 (Rh123), a low toxic fluorescent probe commonly used to assess mitochondrial bioenergetics in living cells, has also been used to measure the efflux activity of P-gp in both normal and malignant cells. Analysis of variation in cellular fluorescence by measuring the rates of Rh123 influx and efflux, together with the effect of mdr reversing agents, allows the investigation of drug-resistant phenotypes in cancer samples. We have studied the functional activity of P-gp in human leukemic cell lines using flow cytometry, taking into consideration that variables such as Rh123 cytotoxicity, culture conditions, cell membrane integrity, as well as the effect of specific P-gp modulators, can impair the resolution of the Rh123-efflux measurements. The studies show that: (1)optimal non-cytotoxic concentrations of Rh123 which allow appropriate color compensation are in the range of 50-200 ng/ml; (2) life-gating allows accurate measurement on the 50% average rate of Rh123 efflux; (3) relative efficiency of P-gp inhibitors was PSC-833 > cyclosporin A > verapamil; and (4) the presence or absence of fetal calf serum had no effect on the bioavailability of chemosensitizer agents, with the exception of serum-free experiments, which showed a significant decrease in P-gp activity under the presence of PSC-833 (P = 0.05). Hence, we recommend this experimental strategy for clinical practice better to study the cellular drug resistance phenotype. PMID- 9205002 TI - Characterization of functional assays of multidrug resistance P-glycoprotein transport activity. AB - P-glycoprotein-mediated multidrug resistance has emerged as one of the most attractive targets to improve anticancer therapy. The P-glycoprotein functions as an energy-dependent, membrane transport pump capable of decreasing the intracellular concentration of a broad range of chemotherapeutic agents. Pharmaceuticals which inhibit P-glycoprotein transport activity are currently being evaluated in clinical trials. Characterization of P-glycoprotein functional activity is critical in determining if these multidrug resistance reversal agents improve therapeutic responses of tumors expressing P-glycoprotein. In this report, we directly compare and characterize assays using rhodamine 123, dimethyloxadicarbocyanine iodide (DiOC2), [3H]daunorubicin and hexakis(2 methoxyisobutyl isonitrile)technetium(I) ([(99m)Tc]Sestamibi) as P-glycoprotein transport probes to quantitate functional activity. The accumulation of certain substrates is concentration dependent and the parameters which determine probe accumulation are impacted by the level of P-glycoprotein expression. In addition, higher concentrations of reversal agents are required to inhibit multidrug resistance in cell lines expressing higher levels of P-glycoprotein. Furthermore, the concentration of reversal agents required to inhibit completely P glycoprotein transport activity is higher than generally recognized. Thus, the level of P-glycoprotein expression may confound intersample comparisons unless sensitive probes are used in combination with saturating concentrations of potent reversal agents. These results highlight the importance of carefully characterizing assay systems under uniform conditions to quantitate P glycoprotein function. PMID- 9205003 TI - Flow cytometric analysis of the multiple drug resistance phenotype. AB - Laser flow cytometry is increasingly used for quantitation of cellular fluorescent drug retention, effect of efflux blockers and for expression of drug resistance related cellular surface markers. Several intrinsic and extrinsic factors can affect the results obtained from drug retention functional assays and lead to artifacts. In the present study, we have used a panel of well characterized parental and drug resistant cell lines, fluorochromes and efflux blockers to identify the possible sources of artefacts in flow cytometric analysis of the multiple drug resistance phenotype. PMID- 9205004 TI - Functional assay of multidrug resistant cells using JC-1, a carbocyanine fluorescent probe. AB - Multidrug resistance (MDR) is characterized by a decrease in the efficiency of chemotherapeutic agents correlated with the expression and activity of a membrane protein: the permeability-glycoprotein (Pgp 170). Clinically, detection of MDR can be performed by functional tests based on the accumulation of fluorescent compounds such as rhodamine 123. With the aim of improving the sensitivity of such analysis, we have evaluated JC-1, a fluorescent lipophilic carbocyanine dye. Above a critical concentration, JC-1 aggregates in a 'liquid crystal' form. Aggregates display a specific red emission band centered at 597 nm whereas the monomers display a green emission band centered at 540 nm. JC-1 was avidly accumulated in sensitive K562 cells where it displayed both a green cytoplasmic and red mitochondrial fluorescence. In contrast, JC-1 was poorly accumulated in resistant K562 cells, which displayed only a slight green fluorescence. The level of JC-1 accumulation was correlated with the level of Pgp expression detected by MRK16 and UIC2 antibodies on a set of K562 subclones with increasing resistance levels. The specific fluorescence properties of JC-1 allow accurate discrimination between low-level resistant cells and sensitive cells. Chemosensitizers such as verapamil, cyclosporine A or S9788 restored JC-1 accumulation in resistant cells. The fluorescence properties of JC-1 could therefore be used for monitoring the effects of reversing agents. PMID- 9205005 TI - Accumulation of simple organic cations correlates with differential cytotoxicity in multidrug-resistant and -sensitive human and rodent cells. AB - Structure/functional studies previously reported showed that in a series of simple organic cations in which the charge is delocalized, an aromatic ring and a minimal degree of lipophilicity (log P > -1) were required for recognition by murine cells which express P-glycoprotein (p-gp)-mediated multidrug resistance (MDR). In the present report we find that 3H-octylpyridinium, the simple aromatic cation which has been shown to be preferentially toxic to MDR- as compared to MDR+ cells, accumulates 4.7-fold greater in the MDR- cell line. In contrast, we find that 3H-guanidinium which displays no selective toxicity between MDR+ and MDR- cells, shows no significant uptake differences between these two cell types. We also present data which demonstrate that other organic cations which contain aromatic rings, a minimal degree of lipophilicity (log P> -1) and carry a delocalized (Rho 123) or shielded (triphenylmethyl phosphonium) positive charge, also accumulate to a greater degree in MDR- vs MDR+ cells. Additionally, we find that human cells which express p-gp MDR, have similar requirements for recognition of these simple compounds. In fact, the sensitivity profiles of these compounds closely correlate between murine and human cell lines. It was also found that none of the series of simple organic compounds tested showed modulatory activity in MDR+ cells, as assayed by monitoring retention of Rho 123. Thus, the requirements for MDR recognition vs those for MDR modulation are clearly distinguished with these simple structured compounds. In comparison, the calcium channel antagonist, verapamil, and a calcium channel agonist, Bay K 8644, both showed modulatory activity by increasing Rho 123 retention in MDR+ cells, further supporting the interpretation that verapamil's modulation of MDR is unrelated to its action on calcium flux. Overall, the data presented here add further information for defining the structural requirements of compounds for their recognition by, or modulation of, human cells expressing p-gp-mediated MDR. PMID- 9205006 TI - Assays for the analysis of P-glycoprotein in acute myeloid leukemia and CD34 subsets of AML blasts. AB - Expression of the multidrug resistance (MDR) phenotype is an independent prognostic variable in acute myeloid leukemia. Approximately 43-57% of the patients have P-glycoprotein (P-gp) expression. A major drawback with the interpretation of P-gp data in AML is the lack of coherence with different analytical assays. We have focused our efforts of P-gp detection on flow cytometry using a dual technique of P-gp staining with antibodies for the extracellular epitope (MRK16) and a functional analysis of P-gp using the rhodamine efflux assay and the effect of P-gp inhibitors such as SDZ PSC 833. This technique was combined with the staining of lineage-specific antigens such as CD34, CD56 and c-kit. In this way, various subsets of AML cells can be identified such as MRK 16+/-, CD34+/- blasts. These cells can be sorted for further analysis, such as the molecular expression of P-gp and other pleiotropic drug resistance genes. PMID- 9205007 TI - Detection of multidrug resistance gene expression in multiple myeloma. AB - Multiple myeloma is a disease which is generally considered responsive to chemotherapy; however, essentially all patients who respond to drug treatment will relapse and die of drug-resistant disease. This disease is therefore considered a paradigm for studying the development of acquired drug resistance in the clinic. Natural product agents are frequently used in the treatment of myeloma, especially vincristine and doxorubicin. Studies using human myeloma cell lines have shown that the MDR1 gene product, P-glycoprotein (Pgp), is responsible for conferring drug resistance to natural products and glucocorticoids. We have developed assays to measure the expression of MDR1/Pgp in human myeloma specimens. These assays include immunocytochemistry, flow cytometry, and RT/PCR. Human myeloma cell lines, 8226/Dox, that are resistant to natural product agents and overexpress MDR1/Pgp are important for standardizing results and offer a means of comparing inter- and intra-patient results. Assays which measure both the presence and function of Pgp are necessary to determine the role of Pgp in clinical drug resistance in patients with myeloma. PMID- 9205008 TI - Anthracycline subcellular distribution in human leukemic cells by microspectrofluorometry: factors contributing to drug-induced cell death and reversal of multidrug resistance. AB - There is a large discrepancy between the changes in drug accumulation and the changes in drug cytotoxicity that accompany development of anthracycline in multidrug-resistant cells. Moreover, although different molecular targets for anthracyclines such as DNA, cell membranes, or enzymes like topoisomerases could be involved, mechanisms by which these compounds exert their cytotoxic and differentiating effects remain unclear. Studies of correlation between the biological effects of anthracyclines and drug uptake have given conflicting conclusions. For example, a decrease in drug cytotoxicity for different incubation temperatures has been observed in spite of the same intracellular anthracycline amount, suggesting that temperature-dependent cytotoxic effects may be mediated by drug interaction with the cell membrane. What we propose in this review are results of our laboratory which are in agreement with an action mechanism targeted to the nucleus. In fact, we have shown by using microspectrofluorometry, that identical nuclear anthracycline concentration induces the same degree of cytotoxicity, independent of cellular MDR phenotype and the anthracycline structure. Thus, we could acquire information on the mechanisms of drug resistance related to drug transport. We could also give evidence that this accumulation is increased when MDR modulators, such as verapamil and S9788 and cyclosporin A or anthracyclines are used. For clinical applications, our studies have already dealt with nuclear concentration measurements of doxorubicin in leukocytes of treated patients, and in vitro measurements of drug efflux from nuclei of acute leukemic cells and its correlation with P-glycoprotein expression. However, in these studies, there was no correlation between anthracycline nuclear accumulation in vitro and P glycoprotein expression. In addition, from preliminary results, we have shown that some modulators such as quinine do not significantly increase nuclear accumulation of anthracyclines in MDR cells but are able to restore anthracycline sensitivity. Other authors have recently shown that quinine has a relatively weak effect on cellular doxorubicin accumulation in MDR cells but is able to completely restore doxorubicin sensitivity. They concluded that quinine has essentially intracellular targets involved in drug distribution (cytoplasm --> nucleus) from sequestration compartments. Our data contradict this and we believe that such modulator modifies the molecular environment of anthracyclines and/or their binding to a possible cytoplasmic target leading to different cell death. Thus, we conclude that mechanisms by which anthracyclines induce cell death, and ways by which chemotherapy fails in resistant cells remain complex and are related to more than one target. PMID- 9205009 TI - Apoptosis and resistance to daunorubicin in human leukemic cells. AB - We compared test methods based on specific mechanisms of daunorubicin (DNR) resistance to more global procedures. Assessment of P-glycoprotein (P-gp) expression and function by means of immunocytochemistry, DNR accumulation, and modulation of resistance and accumulation by the P-gp inhibitor cyclosporin A (CsA) were selected as parameters for multidrug resistance (MDR). On the other hand, we used the MTT assay and measured apoptosis and proliferative activity (S- and G2M-phases of the cell cycle) by flow cytometry. Validation of test methods was achieved for four leukemic cell lines (HL-60, KG-1a, K562/WT, K562/ADM). This battery of tests was then applied to mononuclear cells (MNC) from 18 leukemic patients. Low proficiency of MNC to undergo apoptosis and low proliferative activity rather than P-gp-mediated MDR correlated with DNR resistance as measured by the MTT assay. Bell-shaped dose-response curves for apoptosis, however, which reflect a switch from the apoptotic to the necrotic death mode with increasing cellular damage tend to limit practicability in clinical testing, because appropriate dose range and time points need to be explored. Thus, measurement of apoptosis by flow cytometry may be less convenient than the MTT assay for determination of chemosensitivity, if clinical samples with unknown patterns of responsiveness are to be tested. Spontaneous apoptosis in untreated MNC following 24 h incubation in vitro correlated significantly with DNR sensitivity in the MTT assay. A lack of essential viability factors (eg growth factors or cytokines) in vitro which are known to prevent apoptosis may contribute to DNR sensitivity. PMID- 9205010 TI - The inverse Nehb J lead increases the sensitivity of Holter electrocardiographic monitoring for detecting myocardial ischemia. AB - A major reason for the relatively low sensitivity of Holter electrocardiography (ECG) for detecting ischemia is that the sensitivity of bipolar leads used for Holter ischemia monitoring has not been systematically evaluated, making lead selection difficult. Therefore, this study evaluated the sensitivity of 6 bipolar Holter leads for detecting ischemia during percutaneous transluminal coronary angioplasty. Seventy-five patients, each of whom had > 1 mm ST-segment elevation on an intracoronary electrocardiogram from the myocardium distal to the stenosis during balloon occlusion, were studied for the occurrence of > or = 1 mm ST segment elevation or depression on the simultaneously recorded Holter leads II, III, aVF, CM5, CR4, and inverse Nehb J. The study found that the inverse lead Nehb J provided a significantly higher overall sensitivity for detecting myocardial ischemia than Holter leads II, III, aVF, CM5, and CR4. Also, the use of inverse lead Nehb J significantly increased the sensitivity of 2- and 3-lead Holter ischemia monitoring. These findings were based on a significantly higher sensitivity of inverse lead Nehb J for detecting ischemia induced by transient occlusion of the left anterior descending coronary artery and a slightly higher sensitivity for detecting ischemia induced by occlusion of the left circumflex coronary artery. None of the bipolar leads studied provided a very high sensitivity for detecting ischemia induced by occlusion of the right coronary artery. These findings show that adequate lead selection can increase the sensitivity of Holter ischemia monitoring. Furthermore, the lack of a highly sensitive lead for detection of inferior ischemia indicates that further evaluation of bipolar leads is warranted. PMID- 9205011 TI - Myocardial viability assessed by dobutamine echocardiography in acute myocardial infarction after successful primary coronary angioplasty. AB - Dobutamine echocardiography (5 and 10 microg/kg/ min) was performed in 40 patients 4 +/- 1 days after acute myocardial infarction reperfused by primary coronary angioplasty. The left ventricle was divided into 11 segments. Reversible myocardial dysfunction was indicated by a decrease in at least 2 grades in the total segmental score. Follow-up echocardiography was performed 2 months later. Contractile reserve was documented in 18 patients with dobutamine echocardiography (45%). Sensitivity, specificity, positive, and negative predictive value of dobutamine echocardiography in predicting improvement in contractile function at follow-up were 82%, 83%, 78%, and 86%, respectively. Negative predictive value was high in all dyssynergic segments (86%). Positive predictive value was higher in hypokinetic than in akinetic segments (73% vs 21%; p <0.05). Recovery of wall motion at follow-up was statistically associated with higher left ventricular ejection fraction (p <0.04), collateral blood flow before reperfusion (p = 0.007), and dobutamine responsiveness (p = 0.0001), and was more frequently observed in hypokinetic than in akinetic segments (p <0.05). Thus, low dose dobutamine echocardiography accurately predicts the extent of irreversibly damaged myocardium early after successful direct coronary angioplasty in acute myocardial infarction. PMID- 9205012 TI - Validation of a clinical prediction rule for left ventricular ejection fraction after myocardial infarction in patients > or = 65 years old. AB - We sought to validate a previously described clinical prediction rule for classifying left ventricular ejection fraction (LVEF) after acute myocardial infarction (AMI). As part of the Connecticut cohort of the Cooperative Cardiovascular Project (CCP) pilot study, we identified 3,093 Medicare patients who had been admitted to hospitals throughout Connecticut with an AMI in 1992 and 1993. Retrospective chart review and detailed electrocardiogram interpretation were performed. Of the 1,891 patients with an interpretable EF, 1,378 (73%) had > or = 1 of the rule's exclusion criteria. Of the remaining 513 patients, the clinical prediction rule had a positive predictive value of 89% (i.e., 456 of 513 patients had an EF > or = 40%). In a multivariate model, presentation > 6 hours after the onset of chest pain, a history of bypass surgery, and diabetes mellitus were associated with patients in whom the rule did not correctly predict an EF > or = 40%. Excluding patients with these characteristics from the rule increased the positive predictive value from 89% to 93% and excluded an additional 239 patients. The EF could not be predicted among the patients who did not meet the rule's criteria. In conclusion, a previously published clinical prediction rule for the classification of the EF in patients after an AMI correctly classified 8 of every 9 eligible elderly patients as having an EF > or = 40%. Thus, while not performing as well as it did in the original study, our findings support the use of this rule in providing clinicians with an objective method for estimating an EF > or = 40% in a specific subset of elderly patients. PMID- 9205013 TI - Relation of pacing-induced coronary resistance vessel dilation to total serum cholesterol and heart rate-blood pressure product. AB - Coronary risk factors adversely affect coronary resistance vessel dilation to acetylcholine, but little is known about the effect of risk factors on coronary blood flow (CBF) responses to physiologic stimuli. CBF was derived from Doppler flow velocity (0.018-inch Doppler wire) and coronary diameter (quantitative angiography) in response to rapid atrial pacing in 50 patients (mean age 52 +/- 12 years). Patients were prospectively divided into 3 groups based on their angiograms: group 1 (n = 17), normal coronary arteries; group 2 (n = 18), 1 vessel coronary artery disease (CAD) with a smooth study artery; group 3 (n = 15), 1-vessel CAD and an irregular study artery (<20% stenosis). Pacing produced a significant increase in CBF compared with baseline in groups 1 and 2 (34 +/- 40%, 42 +/- 35%, p < 0.0001), respectively, but not in group 3 (21 +/- 33%), but there was no difference in the pacing response among the 3 groups. The increase in CBF to pacing was inversely related to serum cholesterol (p = 0.01) and triglycerides (p = 0.06) and directly related to the increase in heart rate-blood pressure product (p = 0.007). By multivariate analysis, total cholesterol and the increase in double product were the only factors related to the increase in CBF. Increases in CBF to atrial pacing are inversely related to serum total cholesterol and are not related to the angiographic presence of atherosclerosis in patients with mild CAD. PMID- 9205014 TI - Rescue angioplasty in the thrombolysis in myocardial infarction (TIMI) 4 trial. AB - Rescue percutaneous transluminal coronary angioplasty (PTCA) has been used to establish reperfusion after failed thrombolysis, and the goal of this study was to examine the angiographic and clinical outcomes after rescue PTCA performed for an occluded artery 90 minutes after thrombolysis. Four hundred two patients with acute myocardial infarction were randomized to receive either anistreplase (APSAC), recombinant tissue plasminogen activator, or their combination in the Thrombolysis in Myocardial Infarction (TIMI) 4 trial. The angiographic and clinical outcomes of patients with a patent artery 90 minutes after thrombolysis were compared with those of patients with an occluded artery treated in a nonrandomized fashion with either rescue or no rescue PTCA. At 90 minutes, the number of frames required to opacify standard landmarks (corrected TIMI frame count) was significantly lower (i.e., flow was faster) after successful rescue PTCA (27 +/- 11) than that in patent arteries after successful thrombolysis (39 +/- 20, p < 0.001), and the incidence of TIMI grade 3 flow was correspondingly higher after successful rescue PTCA (87% vs 65%, p = 0.002). In-hospital adverse outcomes (death, recurrent acute myocardial infarction, severe congestive heart failure, cardiogenic shock or an ejection fraction <40%) occurred in 29% of successful rescue PTCAs and in 83% of failed rescue PTCAs (p = 0.01). Among all patients in whom rescue PTCA was performed (successes and failures combined), 35% of patients experienced an adverse outcome, which was the same as the 35% incidence observed in patients not undergoing rescue PTCA (p = NS) and tended to be higher than the 23% incidence observed in patients with patent arteries (p = 0.07). Although successful rescue PTCA for an occluded artery at 90 minutes results in restoration of flow that is superior to that of successful thrombolysis, the incidence of adverse events for the strategy of rescue PTCA as a whole was the same as that of undertaking no PTCA. PMID- 9205015 TI - Coronary vasodilatory capacity and flow reserve in normal myocardium supplied by bypass grafts late after surgery. AB - Coronary artery bypass surgery is used widely for treating myocardial ischemia. However, blood flow and flow reserve of normally perfused myocardium subtended by bypass grafts have not been evaluated late after surgery. Also, it is unknown whether pharmacologic vasodilation evokes comparable myocardial flow responses in arterial and venous conduits. Myocardial blood flow was quantified at rest and during dipyridamole hyperemia using N-13 ammonia and positron emission tomography (PET) in 15 patients 9 +/- 3 years after bypass surgery and in 10 healthy volunteers. Blood flow was analyzed in 26 territories subtended by bypass grafts with normal wall motion and normal perfusion. Myocardial blood flow at rest did not differ between patients and controls (0.65 +/- 0.14 vs 0.68 +/- 0.16 ml/ g/min) and was similar in normal myocardium subtended by saphenous vein (n = 16) and internal mammary artery grafts (n = 10; 0.64 +/- 0.13 vs 0.66 +/- 0.15 ml/g/min). However, the hyperemic response in normal myocardium supplied by bypass grafts was less than that in controls (1.61 +/- 0.33 vs 2.04 +/- 0.30 ml/g/min, p <0.005). No differences between territories supplied by venous and arterial conduits were observed (1.61 +/- 0.35 vs 1.63 +/- 0.32 ml/g/min). Normal myocardium subtended by bypass grafts exhibited a lower flow reserve than that in controls (2.54 +/- 0.51 vs 3.16 +/- 0.85, p <0.02). Myocardial flow reserve was almost identical in regions supplied by venous and arterial grafts (2.55 +/- 0.48 vs 2.52 +/- 0.58). The similar reduction in vasodilatory capacity together with the normal PET polar map findings during dipyridamole argue against flow limiting stenoses in both venous and arterial bypass conduits late after revascularization. Rather, nonobstructive proliferative fibrointimal changes of the bypass conduits or atherosclerosis of the native resistance vessels might account for this finding. PMID- 9205016 TI - Effects of short-term treatment of nicorandil on exercise-induced myocardial ischemia and abnormal cardiac autonomic activity in microvascular angina. AB - The underlying mechanisms of myocardial ischemia in microvascular angina may include endothelial dysfunction, abnormal smooth muscle tone, and abnormal autonomic control of coronary microvasculatures. This randomized, double-blind, placebo-controlled, crossover study was conducted to evaluate the effect of nicorandil (a nitrate-potassium channel opener) therapy on exercise-induced myocardial ischemia and cardiac autonomic activity in 13 patients with microvascular angina. After a 2-week placebo run-in period, patients were randomly assigned to the first 2-week treatment with nicorandil 5 mg tid or placebo, then crossed over to the second 2-week treatment after a 2-week washout period. Treadmill exercise tests and 24-hour ambulatory electrocardiogram monitoring were performed at the end of each treatment phase. The results showed that both time to 1-mm ST depression and total exercise duration were significantly prolonged with nicorandil treatment compared with placebo (p = 0.026 and 0.036, respectively). Maximum exercise ST depression also tended to be less with nicorandil treatment than with placebo (p = 0.083). Compared with 10 healthy control subjects, study patients had significantly reduced heart rate variability in both low- and high-frequency bands while receiving placebo. Nicorandil treatment did not change the altered heart rate variability in either time domain or spectral analysis. Systemic hemodynamics were also unchanged with nicorandil treatment. Thus, 2-week oral nicorandil therapy moderately improved exercise-induced myocardial ischemia without modifying the already altered cardiac autonomic activity, suggesting that nicorandil might have a direct vasodilatory effect on coronary microvasculatures in patients with microvascular angina. PMID- 9205018 TI - Effects of amiodarone on the circadian pattern of sudden cardiac death (Department of Veterans Affairs Congestive Heart Failure-Survival Trial of Antiarrhythmic Therapy). AB - Some antiarrhythmic drugs have been shown to influence the circadian pattern of sudden cardiac death (SCD). The effect of chronic amiodarone therapy on this pattern is unknown. This study determines the circadian pattern of deaths in the Congestive Heart Failure-Survival Trial of Antiarrhythmic Therapy (CHF-STAT) and compares the distribution of SCD between the amiodarone and the placebo arms of the trial. CHF-STAT was a multicenter trial that determined whether amiodarone reduces mortality in patients with heart failure and asymptomatic ventricular arrhythmias. The time of death was retrospectively analyzed in patients who died from pump failure and SCD. In patients who died suddenly, the circadian pattern of deaths was compared between patients receiving amiodarone and those receiving placebo. In CHF-STAT, 274 patients died during follow-up. The time of death was available in 65 of the 74 patients who died from pump failure, and in 96 of the 139 patients who died suddenly. There was a circadian variation of all SCDs compared with other deaths with a distinct peak during the morning (p = 0.04). A similar morning peak of sudden cardiac death was found in both the amiodarone (n = 42) and the placebo (n = 54) groups, and the overall circadian pattern did not differ between them (p = 0.16). In contrast, death from pump failure occurred equally distributed over time. Thus, SCD occurs predominantly during the morning, whereas death from heart failure does not exhibit a morning peak. Amiodarone does not influence the circadian pattern of SCD. PMID- 9205017 TI - A multicenter, double-blind, one-year study comparing safety and efficacy of atorvastatin versus simvastatin in patients with hypercholesterolemia. AB - We directly compared the safety and efficacy of atorvastatin and simvastatin in hypercholesterolemic patients. This 1-year, randomized, double-blind study was performed at 9 community- and university-based research hospitals in Australia. One-hundred seventy-seven patients between the ages of 18 and 80 years with baseline low-density-lipoprotein (LDL) cholesterol > or = 4.14 and < or = 7.76 mmol/L (160 and 300 mg/dl, respectively) and triglycerides < or = 4.52 mmol/L (400 mg/dl) received once-daily dosing with atorvastatin (Lipitor) 10 mg or simvastatin (Zocor) 10 mg. At week 16, the dose of medication was titrated to atorvastatin 20 mg or simvastatin 20 mg if patients did not meet LDL cholesterol target of < or = 3.36 mmol/L (130 mg/dl). Efficacy was reported as percent change from baseline in LDL cholesterol, total cholesterol, very low density lipoprotein cholesterol, total triglycerides, high-density lipoprotein cholesterol, apolipoproteins AI and B, and lipoprotein(a). Atorvastatin caused significantly greater reductions from baseline than did simvastatin for LDL cholesterol, total cholesterol, very low density lipoprotein cholesterol, triglycerides, and apolipoprotein B (p <0.05). No patient in either treatment group had clinically important elevations in creatine phosphokinase, alanine aminotransaminase, or aspartate aminotransaminase. No serious adverse events were considered associated with treatment. With atorvastatin 10 mg, 46% of the patients achieved LDL cholesterol target goal by week 16, whereas only 27% of the simvastatin patients achieved the target goal at the 10-mg dose. This cholesterol-lowering profile affords utility in many patient types. PMID- 9205019 TI - Effect of regular aerobic exercise on elevated blood pressure in postmenopausal women. AB - The efficacy of aerobic exercise for lowering arterial blood pressure (BP) in postmenopausal women with elevations of 130 to 159/85 to 99 mm Hg has not been established. To determine this, 10 postmenopausal women with high normal resting BP or stage I essential hypertension were studied throughout a 12-week lead-in period (no exercise, n = 5) and/or 12 weeks of moderate-intensity aerobic exercise (walking, n = 9). There were no significant time effects during the lead in period (all p >0.4). Maximal aerobic capacity (as assessed by maximal oxygen consumption) was unchanged after 12 weeks of exercise, but exercise tolerance (treadmill walking time) increased by approximately 10% (p <0.05). Body weight, dietary intake and composition, and urinary sodium excretion were unchanged before versus after exercise training. After 12 weeks of exercise, systolic and diastolic BP at rest were significantly lowered by 10/7 and 12/5 mm Hg, respectively, in the sitting and standing positions (p <0.001); some (> or = 3 to 5 mm Hg) decrease in BP was observed in every subject. On average, subjects with stage I hypertension had a reduction in BP into the high normal range, whereas subjects with high-normal initial levels had a reduction in BP into the normal range. Borderline significant (p = 0.06 to 0.07) reductions in systolic and diastolic BP were observed by the end of the second and tenth weeks of training, respectively. Ambulatory determined 24-hour levels of BP were unchanged with training, but significant reductions in BP during submaximal exercise occurred. Our results demonstrate that regular aerobic exercise can produce clinically important reductions in resting BP in Caucasian postmenopausal women with mild to moderately elevated initial levels. This effect of exercise is observed in the absence of changes in maximal aerobic capacity, body weight, or dietary intake. PMID- 9205020 TI - Predictors of response to exercise training in severe chronic congestive heart failure. AB - We prospectively assessed whether baseline central hemodynamics and exercise capacity can predict improvement of VO2 at ventilatory threshold (VT) after exercise training in patients with severe chronic congestive heart failure. Eighteen patients (mean +/- SEM; age 52 +/- 2 years), half of them listed for transplant, underwent 3 weeks of exercise training (interval cycle and treadmill walking; 5 x/week) and 3 weeks of activity restriction in a random-order crossover trial. Baseline data were not significantly different for groups with exercise training first and activity restriction first: cardiac index at rest (2.1 +/- 0.1 L/m2/min), maximum cardiac index (3.1 +/- 0.2 L/m2/min) (Fick), and echocardiographic ejection fraction (21 +/- 1%). The same was true for cardiopulmonary exercise data (cycle ergometry; up 12.5 W/min): VO2 at VT (9.3 +/ 0.4 ml/kg/min), maximum VO2 (12.2 +/- 0.7 ml/kg/min), VT in percentage of predicted maximum VO2 (31 +/- 2%), heart rate at VT (95 +/- 4 beats/min), and decrease of dead space-to-tidal volume ratio from rest to VT (33 +/- 1 --> 29 +/- 1). Improvement of VO2 at VT after training (2.2 +/- 0.4 ml/kg/min; p <0.001) was not related to baseline central hemodynamics (r = <0.10 for each), but was greater in patients with a lower baseline VO2 at VT (r = -0.65; p <0.01), peak VO2 (r = -0.66; p <0.01), VT in percentage of predicted maximum VO2 (r = -0.74; p <0.001), heart rate at VT (r = -0.63; p <0.01), and smaller decrease of dead space-to-tidal volume ratio from rest to VT (r = 0.65; p <0.01). Ejection fraction after exercise training (24 +/- 2%) and activity restriction (23 +/- 2%) did not differ significantly compared with baseline, and patient status (heart failure and cardiac rhythm) remained stable. Three parameters accounted for 84% of the variance of improvement in VO2 at VT: VO2 at VT in percent predicted maximum VO2, decrease of dead space-to-tidal volume ratio, and heart rate at VT. The findings suggest that there was a greater increase in VO2 at VT after exercise training in patients with greater peripheral deconditioning at baseline. The improvement was unrelated to central hemodynamics. Clinically stable patients with severe chronic congestive heart failure, potential heart transplant candidates, and those awaiting transplantation may benefit from involvement in a short-term exercise training program. PMID- 9205021 TI - Safety and clinical utility of long-term intravenous milrinone in advanced heart failure. AB - Few data are available on the long-term safety or clinical utility of the inodilator agent milrinone. We designed a prospective, nonrandomized, observational trial in a cohort of 71 patients who had demonstrated dependence on inotropic therapy, had been clinically stable on an inotropic regimen (milrinone, dobutamine, or both) for > or = 72 hours, and had been given intravenous milrinone for > 72 hours. Group I (n = 22) patients required treatment with both milrinone and dobutamine to achieve stability; group II (n = 49) patients attained stability initially with either milrinone (subgroup IIA) or dobutamine (subgroup IIB), but later required adjunctive therapy with the other inotropic agent for continued hemodynamic support. Of the 71 patients, 38% required mechanical intervention to achieve hemodynamic stability, and 68% were successfully bridged to heart transplantation. Patients were maintained on milrinone therapy for as long as 8 weeks and demonstrated a low incidence of adverse cardiac (7%) or noncardiac (4%) events. Subgroup IIA (28%) had significantly less need than subgroup IIB (52%) for mechanical intervention using an intraaortic balloon pump (p = 0.05), although mortality rates while awaiting transplantation were statistically similar in subgroups IIA (28%) and IIB (35%). Significant improvements from baseline values were noted at the time of transplantation for all aspects of systemic hemodynamics, indicating sustained long-term hemodynamic effects. Long-term intravenous milrinone therapy is safe and well tolerated, and it provides hemodynamic and metabolic support as a pharmacologic bridge to transplantation. The findings also suggest that milrinone as primary inodilator therapy may be associated with less need for mechanical ventricular support. PMID- 9205022 TI - Comparison of left ventricular responses to the six-minute walk test, stair climbing, and maximal upright bicycle exercise in patients with congestive heart failure due to idiopathic dilated cardiomyopathy. AB - Submaximal exercise tests have been advocated to assess exercise capacity in chronic heart failure, but hemodynamic responses have not been characterized. To determine left ventricular (LV) responses during submaximal exercise, the LV ejection fraction (EF) and volumes were evaluated by using an ambulatory radionuclide detector in 13 patients with idiopathic dilated cardiomyopathy during upright maximal graded bicycle exercise, stair climbing and a 6-minute walk test. The 3 tests elicited different responses in volumes and, to a lesser degree, in LVEF. The maximal bicycle exercise led to a decrease in LVEF from 22 +/- 9% to 17 +/- 8% (p <0.05), with marked increases in both end-diastolic volume (EDV) (+15 +/- 10%, p <0.001) and end-systolic volume (ESV) (+23 +/- 18%, p <0.001). Stair climbing tended to reduce LVEF (from 24 +/- 11% to 21 +/- 10%, p = 0.05), with a lesser increase in volumes, which was more marked for ESV (+8 +/- 9%, p <0.01) than for EDV (+4 +/- 4%, p <0.01). The 6-minute walk test did not significantly change LVEF (23 +/- 10% vs 22 +/- 10%), but increased both EDV (+10 +/- 6%, p <0.001) and ESV (+8 +/- 8%, p <0.01) moderately and proportionally. Exercise capacity indexes (peak oxygen consumption, maximal bicycle work rate, stair climbing time, and the distance covered during the 6-minute walk test) correlated significantly with one another. There was no correlation between submaximal exercise tolerance indexes and resting or exercise LVEF. This study shows that (1) LVEF changes are inadequate to report on LV volume changes during exercise; (2) the 3 tests induce different LV volume changes; (3) the 6-minute walk test induces significant changes in LV volumes but no change in LVEF. PMID- 9205023 TI - Expected hemoglobin decrease following percutaneous transluminal coronary angioplasty. AB - To establish expected changes in hemoglobin during and after percutaneous transluminal coronary angioplasty (PTCA), we measured hemoglobin before, at the end of, and on the 2 mornings after PTCA in 177 consecutive patients without obvious out-of-laboratory blood loss. From these data, we calculated confidence intervals that can be used to compare group data, possibly to identify excessive blood loss with new devices or antithrombotic agents, and prediction intervals to identify unexpected blood loss in an individual patient. PMID- 9205025 TI - De novo monomorphic and polymorphic ventricular tachycardia following coronary artery bypass grafting. AB - We compared de novo monomorphic and polymorphic ventricular tachycardia (VT) occurring after coronary artery bypass graft surgery in 21 patients. Our findings support an underlying arrhythmogenic substrate for de novo monomorphic VT, whereas polymorphic VT is more likely related to transient perioperative abnormalities. PMID- 9205024 TI - Routine platelet transfusion in patients undergoing emergency coronary bypass surgery after receiving abciximab. AB - Abciximab has been shown to reduce the ischemic complications of high-risk angioplasty procedures. The appropriate management of patients who have received abciximab and require emergency coronary artery bypass surgery after failed coronary angioplasty is as yet undetermined. We present the outcomes of a small series of such patients who were given platelet transfusions before or during cardiopulmonary bypass. PMID- 9205026 TI - Effect of fluvastatin on low-density lipoprotein peak particle diameter. AB - The effect of fluvastatin on low-density lipoprotein (LDL) particle diameter was investigated in 42 hypercholesterolemic patients. Fluvastatin reduced LDL cholesterol significantly but had no effect on LDL particle diameter; it also had no differential effect on patients classified as LDL pattern A (large LDL), pattern B (small LDL), or I (intermediate LDL). PMID- 9205027 TI - Relation between left ventricular remodeling and nocturnal blood pressure in the elderly with systemic hypertension. AB - Elderly hypertensive patients with concentric left ventricular (LV) hypertrophy had larger LV mass, more impaired diastolic LV function, higher nocturnal blood pressure (BP), and smaller nocturnal BP reduction than those of patients with other geometric patterns. Patients with concentric LV hypertrophy might have more severe hypertensive cardiac involvement and greater risk for cardiovascular events than those patients with other geometric patterns. PMID- 9205028 TI - Comparison of peak exercise oxygen uptake in men versus women in chronic heart failure secondary to ischemic or idiopathic dilated cardiomyopathy. AB - Results of exercise testing in 150 patients with chronic heart failure show that women were characterized by shorter exercise time, peak oxygen consumption, and lower peak oxygen pulse than men. There was a 4.1-ml/kg/min difference in peak oxygen uptake between genders after the adjustment of age, peak heart rate, respiratory exchange ratio, ejection fraction, and etiology of heart failure. PMID- 9205029 TI - Detectable serum troponin I in patients with heart failure of nonmyocardial ischemic origin. AB - Cardiac troponin I, a specific and sensitive marker of myocardial damage, was detected in the blood of 6 of 26 patients studied in our Heart Failure Clinic. In these patients functional class, ventricular function, and prognosis were significantly worse than in those without detectable troponin I. This study suggests that troponin I may represent the biochemical marker of myocardial damage occurring in severe heart failure. PMID- 9205030 TI - Effect of reduced muscle bulk on the ventilatory response to exercise in chronic congestive heart failure secondary to idiopathic dilated and ischemic cardiomyopathy. AB - Changing the exercising muscle group alters the ventilatory response to exercise in chronic heart failure. The recognized muscle abnormalities in congestive heart failure may thus contribute to the ventilatory abnormalities of this condition. PMID- 9205031 TI - Endomyocardial biopsy-proven light chain amyloidosis (AL) without echocardiographic features of infiltrative cardiomyopathy. AB - Amyloid cardiomyopathy may exist when echocardiography does not suggest infiltration. Clinicians should be alert for the presence of a monoclonal protein, nephrotic range proteinuria, or peripheral neuropathy in a patient with heart failure. PMID- 9205032 TI - Clinical significance of fossa ovalis membrane aneurysm in adults with cardioembolic cerebral ischemia. AB - Fossa ovalis membrane aneurysm was diagnosed by transesophageal echocardiography in 45 of 134 consecutive patients (34%) with embolic cerebrovascular ischemic events. A potential cardiovascular source of embolism, other than the fossa ovalis membrane aneurysm, was found in 91% of these patients (41 of 45). PMID- 9205033 TI - Usefulness of combined color Doppler/contrast in providing complete delineation of left ventricular cavity. AB - Contrast-enhanced 2-dimensional echocardiography without color Doppler did not result in complete filling of the left ventricular cavity in 21 patients studied. However, contrast-enhanced color Doppler was very effective and provided complete opacification of the left ventricular cavity in 20 of these 21 patients. PMID- 9205034 TI - Computer treason: intraobserver variability of an electrocardiographic computer system. AB - An electrocardiogram computer system significantly disagreed with itself in 36 of 92 pairs of unselected electrocardiograms which had not changed when recorded 1 minute apart. PMID- 9205035 TI - Prospective evaluation of a stiff shaft glide wire compared with the standard straight wire in crossing severely stenotic aortic valves. AB - In this prospective randomized study of the use of the Terumo glide wire compared with the standard straight wire for crossing of severely stenotic aortic valves, the glide wire was shown to significantly decrease the fluoroscopy time of the procedure and to lower by 3.4 times the need for crossover to the alternative technique. PMID- 9205036 TI - The rule of 5 and the rule of 7 in lipid-lowering by statin drugs. PMID- 9205037 TI - Eventual recovery of regional perfusion after acute myocardial infarction. PMID- 9205038 TI - Dipyridamole-induced myocardial ischemia. PMID- 9205039 TI - QT hysteresis and syncope. PMID- 9205040 TI - What is ESMIR? PMID- 9205041 TI - Organization of efferent projections from the parabrachial area to the hypothalamus: a Phaseolus vulgaris-leucoagglutinin study in the rat. AB - The organization of projections from the parabrachial (PB) area to the hypothalamus was studied in the rat by using microinjections of Phaseolus vulgaris-leucoagglutinin (PHA-L) into subregions of the PB area. The present study is a follow-up of two former studies (Bernard et al. [1993] J. Comp. Neurol. 329:201-229; Alden et al. [1994] J. Comp. Neurol. 341:289-314) that examined PB projections onto the amygdala and the bed nucleus of the stria terminalis. The results demonstrate that 1) the mesencephalic PB region, centered in the lateral portion of the superior lateral subnucleus projects extremely densely to almost the entire dorsomedial subdivision of the ipsilateral ventromedial hypothalamic nucleus; 2) the mesencephalic PB region, located in the medial portion of the superior lateral subnucleus and weakly overflowing into the rostralmost dorsal lateral pontine subnucleus, projects densely to the retrochiasmatic area and, to a lesser extent, to the ipsilateral ventromedial nucleus of the hypothalamus; 3) the PB region, including the central lateral, a portion of the superior lateral, and the outer external lateral subnuclei, projects densely to the ipsilateral median, anteroventral, and periventricular preoptic hypothalamic nuclei and projects more weakly to the dorsal border of the paraventricular nucleus (PVN). No consistent projection was found in the magnocellular PVN. All of these PB regions also project diffusely to the dorsomedial area and to a small tuberal subfornical hypothalamic area. In addition, the medial half of the PB area projects consistently to the posterior lateral hypothalamus. It is suggested that these pathways may be involved in aversive-defensive behavior, in autonomic and neuroendocrine aspects of pain, and in feeding and energy metabolism regulation. PMID- 9205042 TI - The relevance of neural architecture to visual performance: phylogenetic conservation and variation in Dipteran visual systems. AB - In cyclorrhaphan flies, giant tangential neurons in the lobula plate are supplied by isomorphic arrays of evolutionarily conserved achromatic elementary motion detecting circuits originating in the retina. The arrangements among giant tangential neurons is characteristic of a taxon and can differ between taxa having different visual performances. Observations of 12 brachyceran and 4 nematoceran species have identified different behaviors associated with visually stabilized flight. Neuroanatomical comparisons between closely related species having different behaviors and phylogenetically distant species that have similar behaviors suggest that such differences relate to differences of giant tangential cell architecture in the lobula plate. These functionally related differences contrast to anatomical features that reflect phylogenetic affinities. For example, the lobula plates of robber flies, typified by ballistic flight behavior, all differ from other taxa in lacking cyclorrhaphan-type vertical motion-sensitive neurons; instead, they possess an extra complement of horizontal cells in their place. The results suggest that, although circuits that compute elementary motion are conserved across the Diptera, selective pressure has resulted in modifications of their target neurons, thus contributing to the wide variety of visual behaviors observed within this group of insects. PMID- 9205043 TI - Distribution of oxytocin- and vasopressin-binding sites in the rat extended amygdala: a histoautoradiographic study. AB - Radioligand receptor autoradiography has shown that oxytocin- and vasopressin binding sites exist in numerous rat brain regions, among which the amygdala and the bed nucleus of the stria terminalis (BST) are especially prominent. However, these descriptions did not take into account the numerous subdivisions of the amygdala and the BST. Thus, we have reinvestigated the distribution of these sites in the rat extended amygdala, which is formed by a continuum of structures stretching from the BST to the centromedial amygdala, including parts of the accumbens nucleus, substantia innominata, and transition areas between the amygdala and the striatum. For this purpose, histoautoradiography was used to detect binding sites at the cellular level, and anatomical boundaries were defined on the basis of acetylcholinesterase histochemistry and tyrosine hydroxylase immunohistochemistry. Oxytocin- and vasopressin-binding sites were detected in well-defined subdivisions of both medial and central parts of the extended amygdala, but they almost never coexisted in the same region. Compared with previously reported distributions, our reinvestigation describes novel oxytocin- and vasopressin-binding sites in the lateral and supracapsular BST, in the sublenticular extended amygdala, in the interstitial nucleus of the posterior limb of the anterior commissure, in the marginal zone, in the central amygdaloid nucleus, and in the anterior amygdaloid area. These results indicate that oxytocin- and vasopressin-binding sites represent an important feature of the extended amygdala and may participate in the large variety of functions that characterize this area, including reproductive and ingestive behaviors, conditioned fear and autonomic regulation. PMID- 9205044 TI - Postnatal development of terminals and synapses in laminae I and II of the rat medullary dorsal horn. AB - To better understand developing orofacial nociceptive circuits and to provide a baseline for evaluating injury-induced plasticity, the ultrastructure of the superficial laminae in the rat medullary dorsal horn was examined at birth and at postnatal days 1, 4, 17, and 90. Quantitative features of terminals and synapses were studied with stereological methods. In laminae I and II: 1) Axon terminal density increased significantly from birth to day 4 and again from day 4 to day 90. 2) The density of degenerating profiles increased significantly from birth to day 1 and from birth to day 4 and then decreased from day 4 to day 90. 3) Degenerating profiles were most dense on day 1 and declined steadily thereafter; by day 90, such profiles were rare. 4) Cavitation was by far the most common form of degeneration seen at early postnatal ages. 5) Growth cone-like profiles were most dense at birth and declined steadily during the first 2 postnatal weeks; by day 90, such profiles were absent. 6) Terminals with flat synaptic vesicles were rarely seen before day 90, when they accounted for 7% of the terminal population. 7) The density of synapses increased continuously from birth until day 90. These data suggest that, as in the spinal cord, medullary dorsal horn circuits are very immature at birth. Adult-like quantitative features are not attained until after day 17. Moreover, whereas degenerating profiles are prevalent during early postnatal development, and they have features that resemble naturally occurring degeneration, the total numbers of terminals and synapses continue to increase dramatically and gradually during a protracted postnatal period (to postnatal day 17). PMID- 9205045 TI - Development of terminals and synapses in laminae I and II of the rat medullary dorsal horn after infraorbital nerve transection at birth. AB - Infraorbital nerve damage at birth kills neurons and alters anatomical, physiological, and biochemical properties of surviving cells in all portions of the trigeminal brainstem complex, with the exception of laminae I and II of the medullary dorsal horn. The resiliency of laminae I and II may be due to rapid terminal sprouting and reactive synaptogenesis in this region. To test this hypothesis, quantitative electron microscopy revealed the types and numbers of terminals, synapses, and degenerating and growth cone-like profiles in the left laminae I and II at 1, 4, 17, and 90 days after left infraorbital nerve section. Control data were derived from normal newborns and from the right laminae I and II and the left infraorbital nerve of every experimental animal. Deafferented laminae I and II contained a median of 11.7, 8.2, 21.8, and 38.2 synapses/100 microm3 on days 1, 4, 17, and 90, respectively. At corresponding ages, there were 17.1, 19.4, 36.2, and 32 terminals; 14.4, 4.2, 5.1, and 0.3 degenerating profiles; and 4.6, 2.2, 0.1, and 0 growth cone-like profiles/100 microm2. Significant differences from the control right side are: 1) The percentage area occupied by terminals is less on days 1 and 17; 2) terminal density does not increase from day 0 to day 4 as it does on the control side; 3) the density of degenerating profiles is higher on day 17; 4) growth cones are less dense on days 4 and 17; and 5) synapse density is lower on days 1 and 4. Axon number in the infraorbital nerve was highly predictive of terminal and synapse densities in deafferented laminae I and II at all ages. Thus, in laminae I and II, 1) the time course and nature of development are altered by deafferentation at birth; 2) reorganization of terminals and synapses occurs within a day of the lesion; 3) by day 90, there are no remaining lesion effects; and 4) the status of the injured nerve predicts central terminal and synapse densities. These are signs of injury induced transganglionic degeneration and sprouting. The source of the latter is unknown, although areal fraction data suggest that "replacement" terminals may not be of primary afferent origin. PMID- 9205046 TI - Basal ganglia organization in amphibians: development of striatal and nucleus accumbens connections with emphasis on the catecholaminergic inputs. AB - To broaden our insight into the organization of the basal ganglia of amphibians, the development of the connections of the striatum and the nucleus accumbens was studied by means of tract-tracing techniques based on the transport of biotinylated dextran amines. In a number of experiments, these techniques were combined with tyrosine hydroxylase immunohistochemistry to identify the sources of catecholaminergic inputs to the striatum and the nucleus accumbens. Already at late embryonic stages, the basal telencephalon receives inputs from cells located in the amygdala, the thalamus, the suprachiasmatic nucleus, the raphe nucleus, and the rhombencephalic reticular formation. At these stages, the rostral part of the posterior tubercle seems to be the only source of the dopaminergic input to the basal telencephalon. During premetamorphosis, not only a differentiation between connections of the striatum and the nucleus accumbens could be made, but new sources of inputs were also detected in the mesencephalic and isthmic tegmentum, the parabrachial nucleus, and the nucleus of the solitary tract. Double-labeling experiments revealed that, at these stages, in addition to the posterior tubercle, cells within the mesencephalic tegmentum, the locus coeruleus, and the solitary tract nucleus contribute to the catecholaminergic innervation of the basal forebrain. During prometamorphic stages, a gradual increase occurs in the number of cells that project to the basal telencephalon. At the beginning of the metamorphic climax, the organization of the basal ganglia afferents largely resembles the pattern observed in juveniles and adults. Remarkably, during larval stages, the cells that contribute to the dopaminergic innervation of the basal forebrain show a rostrocaudal gradient in time of appearance. Moreover, the dopaminergic fibers reach the striatum earlier than the nucleus accumbens, and they precede markedly the development of the efferent connections of both brain structures. These developmental aspects are easily correlated with the situation in amniotes; therefore, the notion that amphibians share an essentially similar pattern of basal ganglia organization with other tetrapods is further strengthened. PMID- 9205047 TI - Corticospinal tract neurons are radially malpositioned in the sensory-motor cortex of the Shaking rat Kawasaki. AB - Shaking rat Kawasaki (SRK) is an autosomal recessive mutant rat that exhibits tremor, dystonia, and ataxia and that is characterized by abnormal lamination of the cerebral and cerebellar cortices and the hippocampus. To examine whether or not layer V neurons in the mutant neocortex are malpositioned in accordance with the aberrant laminar cytoarchitecture, horseradish peroxidase (HRP) was injected into the lumbar spinal cord of SRK mutant and normal control rats to label cortical pyramids projecting through the corticospinal tract (CST). HRP-labeled CST neurons of both normal and SRK rats were found mainly in the hindlimb area of the sensory-motor cortex, indicating a normal tangential distribution of labeled CST neurons in the SRK mutant. In the radial axis, however, labeled CST neurons were spread throughout all layers of the mutant cortex, whereas those in normal rats were restricted to layer V. In the mutant, most labeled CST neurons located in the inner third of the cortex had a typical pyramidal form with an upright apical dendrite, but many of those located near the pial surface displayed abnormal shapes and could be subdivided into inverted pyramidal, horizontal, and bipolar neurons on the basis of their dendritic morphology. The abnormal distribution pattern of labeled CST neurons in the mutant was quantified using a standardized measure of their depth distribution, where 0% = the level of the white matter and 100% = the pial surface. The mean value for the SRK cortex of 47.0% was significantly greater than the figure of 40.5% for normal rats (P < 0.01, Student's t test), indicating a spread of CST neurons toward the pial surface in SRK, but even more striking was the size of the standard deviation: 30.4 in SRK compared with 7.1 in controls. The distribution pattern of CST neurons of the SRK rat was also statistically identical with that of the reeler mouse, which is a well-known mutant that also exhibits an abnormal lamination pattern in the cerebral cortex. These results indicate that neuronal components of the neocortex of the SRK mutant are intermingled along the radial axis, but not in the tangential axis, and provide further evidence for a strong similarity between this spontaneous rat mutation and the reeler malformation. PMID- 9205048 TI - Glutamate-positive neurons and terminals in the cat periaqueductal gray matter (PAG): a light and electron microscopic immunocytochemical study. AB - The morphology, distribution, proportion, size, and synaptic organization of periaqueductal gray matter neurons labeled with immunocytochemical techniques by an anti-glutamate (Glu) polyclonal serum were investigated in six adult cats (PAG GLU 1-6). At the light microscopic level, numerous Glu-positive neurons were found throughout each subdivision of the periaqueductal gray matter. Their proportion and size, calculated in semi-thin sections (1-microm-thick), varied slightly among the subdivisions of the periaqueductal gray matter. The morphology of Glu-positive neurons was similar to that of the multipolar, triangular, and fusiform cells described in previous Golgi studies. Numerous puncta, interpreted as dendrites, axons, and axon terminals were also present in all subdivisions without preferential distribution. At the electron microscopic level, all synaptic contacts made by Glu-positive axon terminals were of the asymmetric type, but not all presynaptic elements making asymmetric synapses were labeled. The vast majority of postsynaptic elements contacted by Glu-positive axon terminals were labeled and unlabeled dendrites. The present results describe for the first time the presence of both Glu-positive neurons and terminals in the feline periaqueductal gray matter and provide further evidence that Glu is the probable neurotransmitter of numerous excitatory neurons of this structure. PMID- 9205049 TI - Deficient transforming growth factor-beta1 activation and excessive insulin-like growth factor II (IGFII) expression in IGFII receptor-mutant tumors. AB - The insulin-like growth factor II receptor (IGFIIR) gene has been identified as a coding region target of microsatellite instability in human gastrointestinal (GI) tumors. IGFIIR normally has two growth-suppressive functions: it binds and stimulates the plasmin-mediated cleavage and activation of the latent transforming growth factor-beta1 (LTGF-beta1) complex, and it mediates the internalization and degradation of IGFII ligand, a mitogen. We used an immunohistochemical approach to determine whether IGFIIR mutation affected expression of these proteins in GI tumors. Four highly specific antibodies were used: LC(1-30), which recognizes the active form of TGF-beta1; anti-LTGF-beta1, which detects the LTGF-beta1 precursor protein; anti-IGFIIR; and anti-IGFII ligand. Twenty GI tumors either with (6 of 20) or without (14 of 20) known IGFIIR mutation were examined, along with matching normal tissues. Results were statistically significant in the following categories: (a) decreased active TGF beta1 protein expression in IGFIIR-mutant tumor tissues versus matching normal tissues or IGFIIR-wild-type tumor tissues; (b) increased LTGF-beta1 protein expression in IGFIIR-mutant tumor tissues versus matching normal tissues or IGFIIR-wild-type tumor tissues; and (c) increased IGFII ligand protein expression in IGFIIR-mutant tumor tissues versus matching normal tissues or IGFIIR-wild-type tumor tissues. These data suggest that in genetically unstable GI tumors, mutation of a microsatellite within the coding region of IGFIIR functionally inactivates this gene, causing both diminished growth suppression (via decreased activation of TGF-beta1) and augmented growth stimulation (via decreased degradation of the IGFII ligand). PMID- 9205051 TI - p53-dependent DNA damage-induced apoptosis requires Fas/APO-1-independent activation of CPP32beta. AB - In many cell types, the p53 tumor suppressor protein is required for the induction of apoptosis by DNA-damaging chemotherapy or radiation. Therefore, identification of the molecular determinants of p53-dependent cell death may aid in the design of effective therapies of p53-deficient cancers. We investigated whether p53-dependent apoptosis requires activation of CPP32beta (caspase 3), a cysteine protease that has been found to mediate apoptosis in response to ligation of the Fas molecule or to granzyme B, a component of CTL lytic granules. Irradiation-induced apoptosis was associated with p53-dependent activation of CPP32beta-related proteolysis, and normal thymocytes were protected from irradiation by Acetyl-Asp-Glu-Val-Asp-CHO (Ac-DEVD-CHO), a specific inhibitor of CPP32beta. We next examined whether the Fas system is required for p53-dependent apoptosis and whether stimuli that induce activation of CPP32beta induce apoptosis in p53-deficient cells. Thymocytes or activated T cells from Fas deficient mice were resistant to apoptosis induced by ligation of Fas or CD3, respectively, but remained normally susceptible to irradiation. Thymocytes from p53-deficient mice, although resistant to DNA damage, remained sensitive to CPP32beta-mediated apoptosis induced by ligation of Fas or CD3, or by exposure to cytotoxic T cells. These results demonstrate that DNA damage-induced apoptosis of T cells requires p53-mediated activation of CPP32beta by a mechanism independent of Fas/FasL interactions and suggest that immunological or molecular methods of activating CPP32beta may be effective at inducing apoptosis in p53-deficient cancers that are resistant to conventional chemotherapy or irradiation. PMID- 9205050 TI - Selective inhibition of estrogen-regulated gene expression in vivo by the pure antiestrogen ICI 182,780. AB - Estrogen-stimulated cell proliferation is thought to be mediated by the induction of growth-regulatory factors. Abnormal regulation of such factors may be associated with tumorigenesis. Two such factors, vascular endothelial growth factor and c-fos, are rapidly induced in the uterus by estrogens. We show that the induction of these transcripts by 17beta-estradiol or tamoxifen is selectively blocked by the pure antiestrogen ICI 182,780 in a dose-dependent manner. This indicates that induction of the two genes requires different levels of estrogen receptor or that their induction occurs by different mechanisms. This suggests that selective dose-dependent antagonism of estrogen-dependent transcription may be possible in target tissues and tumors. PMID- 9205052 TI - Role of interleukin 1 and granulocyte colony-stimulating factor in photofrin based photodynamic therapy of rat rhabdomyosarcoma tumors. AB - Neutrophils play an important role in the efficacy of photodynamic therapy (PDT). These leukocytes rapidly accumulate into the tumor lesion after PDT and most likely eradicate the remaining attenuated tumor cells. The underlying mechanism of the accumulation of neutrophils at the time of PDT is not known. Therefore, we determined the effect of PDT on the course of mature and immature neutrophils in the circulation of rhabdomyosarcoma-bearing rats and studied the changes in the level of interleukin (IL)-1beta as an important stimulator of the proliferation of precursor cells of the granulocyte lineage in the bone marrow. We found that the effect of PDT on tumor growth was preceded by a rapid and specific increase of the number of mature neutrophils in the peripheral blood as early as 4 h after the start of PDT treatment and reaching maximum values after 8 h. At 24 h, the neutrophil numbers in the PDT-treated rats were still elevated as compared to sham-treated rats. In sham-treated rats, the numbers of blood monocytes and lymphocytes decreased by about 50% after 2 h and returned to their normal levels as soon as 2 h later. In PDT-treated rats, the course of monocyte numbers showed a similar pattern; however, lymphocyte numbers did not reach the normal range until 24 h. The specific increment of neutrophils was preceded by an increase of band neutrophil numbers and elevated serum levels of IL-1beta which were maximal at 2 h after the start of PDT. Pearson correlation analysis showed a significant association between the serum levels of IL-1beta at this time point and the number of band neutrophils at 4 h (R2 = 0.58; P = 0.03) and the number of mature neutrophils at 8 h (R2 = 0.54; P = 0.04). This suggests that PDT evoked an IL-1 dependent increased production rate of neutrophils in the bone marrow. Further investigation showed that the injection of anti-granulocyte colony-stimulating factor (G-CSF) antibodies not only attenuated the increase in neutrophil numbers but also greatly decreased the efficacy of PDT. On this basis, we suppose that an IL-1-induced release of G-CSF by PDT underlies this nonspecific immune reaction to the tumor. Apparently, G-CSF not only stimulates the production rate of neutrophils in the bone marrow but also increases the functional activity of these leukocytes to become indispensable tumor cell killers. PMID- 9205053 TI - Prostate attenuated replication competent adenovirus (ARCA) CN706: a selective cytotoxic for prostate-specific antigen-positive prostate cancer cells. AB - Prostate-specific antigen (PSA) is a widely used marker for the diagnosis and management of prostate cancer. Minimal enhancer/promoter constructs derived from the 5' flank of the human PSA gene (prostate-specific enhancer) were inserted into adenovirus type 5 DNA so as to drive the E1A gene, thereby creating a prostate-specific enhancer-containing virus, CN706. E1A was expressed at high levels in CN706-infected human PSA-producing LNCaP cells but not in CN706 infected DU145 cells, which are human prostate cells that do not express PSA. The titer of CN706 was significantly higher in LNCaP cells compared to several human cell lines that do not produce PSA (HBL100, PANC-1, MCF-7, DU145, and OVCAR3). Furthermore, in LNCaP cells, the yield of CN706 was dependent on exogenous androgen (R1881). CN706 destroyed large LNCaP tumors (1 x 10(9) cells) and abolished PSA production in nu/nu mouse xenograft models with a single intratumoral injection. PMID- 9205054 TI - The immunogenicity of experimental tumors is strongly biased by the expression of dominant viral cytotoxic T-lymphocyte epitopes. AB - The immunogenic Friend-Moloney-Rauscher (FMR) virus-induced tumors have been used extensively to clarify the cellular and molecular mechanisms responsible for tumor rejection and to develop immunotherapeutic strategies. We characterize here the trimolecular complex MHC class I-antigenic determinant-T cell receptor involved in the induction of a protective CTL response against the RMA thymoma. This complex is mainly composed by the D(b) molecule interacting with a Rauscher virus antigen (Ag) determinant and the Vbeta5+ T cell receptor. We also show that the chemically induced EL-4 thymoma acquires the susceptibility to recognition by anti-RMA CTLs and the ability to elicit a protective anti-RMA CTL response only upon infection by a virus of the FMR family and that RMA and FMR virus infected EL-4 cells share tumor-associated Ag. The data strongly support the hypothesis that the high immunogenicity of virus-induced or infected tumors is determined by the expression of immunodominant virus-encoded Ag. The demonstration of a different outcome in the immune responses elicited in the presence or in the absence of viral Ag further open the contention of the molecular requirements for immunogenicity and should stimulate a more careful revision of unexpected cross reactivity among tumors. PMID- 9205055 TI - Protective immunity induced by tumor vaccines requires interaction between CD40 and its ligand, CD154. AB - Interactions between CD40 and its ligand, CD154 (CD40L, gp39), have been shown to play a central role in the regulation of humoral immunity. Recent evidence suggests that this ligand-receptor pair also plays an important role in the induction of cell-mediated immune responses, including those directed against viral pathogens, intracellular parasites, and alloantigens. The contribution of this ligand-receptor pair to the development of protective immunity against syngeneic tumors was evaluated by blocking the in vivo function of CD154 or by studying tumor resistance in mice genetically deficient in CD40 expression (CD40 /-). In the former case, anti-CD154 monoclonal antibody treatment inhibited the generation of protective immune responses after the administration of three potent tumor vaccines: irradiated MCA 105, MCA 105 admixed with Corynebacterium parvum adjuvant, and irradiated B16 melanoma cells transduced with the gene for granulocyte macrophage colony-stimulating factor. Confirmation of the role of CD40/CD154 interactions in tumor immunity was provided by the overt tumor susceptibility in CD40-deficient mice as compared to that in CD40+/+ mice. In this case, wild-type but not CD40-deficient mice could be readily protected against live TS/A tumor challenge by preimmunization with TS/A admixed with C. parvum. These findings suggest a critical role for CD40/CD154 interactions in the induction of cellular immunity by tumor vaccines and may have important implications for future approaches to cell-based cancer therapies. PMID- 9205056 TI - Loss of imprinting of the insulin-like growth factor II gene in renal cell carcinoma. AB - Loss of imprinting (LOI) has been implicated in the pathogenesis of embryonal malignancies as well as adult cancers. Insulin-like growth factor II (IGF2) gene is an imprinted gene, normally transcribed only from the paternal allele. We investigated allele-specific expression of the IGF2 gene in 22 cases of renal cell carcinoma (RCC), a common adult-onset renal tumor. Sixteen cases (72%) were informative for IGF2 gene expression, and 9 (56%) of these cases showed biallelic expression of the IGF2 gene. Additionally, in four cases with biallelic expression from which uninvolved kidney tissue was available, LOI of the IGF2 gene was also demonstrated in the normal tissue. All cases with LOI of IGF2 were low-grade and low-stage tumors. LOI of the IGF2 gene in RCC was not associated with overexpression of IGF2 mRNA, whereas IGF2 overexpression was frequently observed in high-stage tumors. These results suggest that LOI of IGF2 predisposes to low-grade and low-stage tumors, whereas IGF2 overexpression may have a role in RCC tumor progression. PMID- 9205057 TI - Frequency of Smad gene mutations in human cancers. AB - Much excitement has recently been generated by the discovery of the Smad genes, encoding proteins that transduce signals from the transforming growth factor beta family of cytokines. Here, we report the completion of cloning of the six known human Smads, providing novel sequences for Smad5 and Smad6. Previously, Smad4 and Smad2 were shown to be mutated in human cancers. However, analysis of the other four Smad genes revealed no mutations in a total of 167 tumors, including those from colon, breast, lung, and pancreas. These results suggest that the various Smad genes have different functions and demonstrate that mutations in these four genes do not, in general, account for the widespread resistance to transforming growth factor beta that is found in human tumors. PMID- 9205058 TI - Mutations in the human homologue of the Drosophila segment polarity gene patched (PTCH) in sporadic basal cell carcinomas of the skin and primitive neuroectodermal tumors of the central nervous system. AB - The human homologue of the Drosophila segment polarity gene patched (PTCH) has recently been identified as the tumor suppressor gene responsible for the nevoid basal cell carcinoma (BCC) syndrome (H. Hahn et al., Cell, 85: 841-851, 1996; R. L. Johnson et al., Science (Washington DC), 272: 1668-1671, 1996). In addition to multiple BCCs, patients with nevoid BCC syndrome have a predisposition for the development of primitive neuroectodermal tumors (PNETs) of the central nervous system. We have analyzed 9 sporadic BCCs and 37 PNETs for mutation and expression of the PTCH gene. PTCH mutations were found in 3 BCCs (33.3%) and in 5 PNETs (14%), including 1 of 5 cerebral PNETs, 2 of 15 medulloblastomas, and 2 of 17 desmoplastic medulloblastomas. The sequence changes in six of these tumors (four PNETs, two BCCs) were mutations predicted to result in truncated proteins. Missense mutations were detected in one PNET and one BCC each. In addition, novel sequence polymorphisms were found in exon 2, intron 5, intron 10, and intron 14 of PTCH. Reverse transcription-PCR analysis revealed increased PTCH expression levels compared to nonneoplastic brain tissue and normal skin in the majority of PNETs and BCCs investigated. Our data suggest that genetic alterations of PTCH are not only of significance in hereditary and sporadic BCCs but are also involved in the molecular pathogenesis of a subset of sporadic central nervous system PNETs. PMID- 9205059 TI - Effect of folate deficiency on mutations at the hprt locus in Chinese hamster ovary cells exposed to monofunctional alkylating agents. AB - Multiplex PCR amplification of hprt exons from 113 Chinese hamster ovary cell clones selected for resistance to 6-thioguanine was performed to investigate the molecular basis for the synergistic mutagenic effects of nutritional folic acid deficiency and alkylating agents. In cells treated with ethyl methanesulfonate, intragenic deletions were detected in 9 of 46 (19.6%) clones derived from folate deficient cells, but in none of 16 mutants grown in folate-replete medium. The number of deletions found in mutants generated by N-nitroso-N-ethylurea was low in both folate-deficient (1 of 25; 4%) and folate-replete (1 of 26; 3.8%) cells. Correction of folate deficiency may decrease the frequency of intragenic deletions caused by some alkylating agents. PMID- 9205060 TI - Comparisons of CYP2D messenger RNA splice variant profiles in human lung tumors and normal tissues. AB - Allelic variants of the CYP2D6 gene, a member of the cytochrome P450 gene superfamily, have been implicated in susceptibility to lung carcinogenesis. Human breast CYP2D6 and CYP2D7P (from a pseudogene) mRNAs were previously reported to be expressed as a series of splice variants. In this study, the expression of full-length and splice variants of these mRNAs in human lung tissue and tumors are reported for the first time and are compared in order to probe the potential for differential CYP2D6 regulation in lung normal tissue and tumors. The splice variant profiles differed within the same individual, but no consistent differences were detected. PMID- 9205061 TI - Differential display cloning identifies motility-related protein (MRP1/CD9) as highly expressed in primary compared to metastatic human colon carcinoma cells. AB - Differential display cloning was performed to analyze genes that are differentially expressed in matched primary and metastases-derived human colon carcinoma cell lines. This led to the identification of PMA16, a gene identical to the previously cloned motility-related protein gene (MRP1/CD9). Northern and Western blot analyses of cell lines, as well as immunostaining of tissue sections from the original tumor surgical samples, confirmed that MRP1/CD9 was highly expressed at the primary site, compared to the low levels of expression in metastases. We also demonstrated that primary colon cancer cells displayed a significantly higher migration potential, compared to metastasis-derived cells. Antibodies directed against MRP1/CD9 largely prevented cell migration in vitro, but they did not influence cell adhesion. Thus, differential display cloning has allowed for the identification of MRP1/CD9, a motility-related gene product, which may regulate the metastatic phenotype of human colon cancer. PMID- 9205062 TI - 5-fluorouracil kinetics in the interstitial tumor space: clinical response in breast cancer patients. AB - Several anticancer drugs fail to exhibit sufficient activity against solid tumors in vivo despite effective inhibition of tumor cell growth in vitro. This may be due to impaired drug transfer from plasma into solid tumors. The present study, therefore, aimed at measuring interstitial tumor 5-fluorouracil (5-FU) pharmacokinetics and 5-FU transfer rates from plasma into the tumor interstitium in breast cancer patients. Microdialysis probes were inserted into the primary tumor and the periumbilical s.c. adipose layer of 10 breast cancer patients (8 females and 2 males) scheduled to receive neoadjuvant chemotherapy due to locally advanced breast cancer. Thereafter, patients received 5-FU (600 mg/m2, i.v). 5-FU kinetics were followed in plasma and tumor and s.c. interstitial fluid. Mean interstitial 5-FU load, expressed as area under curve (AUC), in breast tumors was 61 +/- 11% (means +/- SE) of the mean plasma 5-FU load. 5-FU displayed similar kinetics in the interstitial space of s.c. adipose tissue and tumor tissue. A high interstitial tumor AUC was associated with increased tumor response. There was no association with tumor response for s.c. or plasma AUC of 5-FU. Measurement of interstitial drug concentrations in breast tumors by in vivo microdialysis may predict response to chemotherapy. This information may explain drug resistance in some patients and help to optimize dosing and administration schedules. In the future, selection of novel cytotoxic compounds with favorable tumor penetration characteristics may become possible. PMID- 9205063 TI - The extracellular fluid of solid carcinomas contains immunosuppressive concentrations of adenosine. AB - The purine nucleoside adenosine (9-beta-D-ribofuranosyladenine) inhibits a number of lymphocyte functions in vitro, including the ability of activated T lymphocytes and natural killer cells to adhere to and kill tumor targets. Solid tumors, such as adenocarcinomas of the lung and colon, are frequently hypoxic and are, therefore, likely to exhibit increased adenine nucleotide breakdown through the 5'-nucleotidase pathway, yielding adenosine. We examined whether the concentration of adenosine in the extracellular fluid of such tumors is adequate to cause immunosuppression. Murine tumors grown in syngeneic hosts or human tumors grown in immunodeficient nu/nu mice were subjected to microdialysis, and adenosine levels in the microdialysate were measured by high-performance liquid chromatography. Treatment of the tumor microdialysates with adenosine deaminase eliminated the adenosine peak. Recovery of adenosine ranged from 15 to 29%, depending on the microdialysis probe, and concentrations of adenosine in tumors ranged from 0.2 to 2.4 microM with a mean of 0.5 microM. In contrast, the adenosine concentration measured s.c. at the same location was 30 +/- 5 nM (mean +/- SE). Inclusion of the adenosine deaminase inhibitor coformycin (10 microM) and the adenosine kinase inhibitor 5'-iodotubercidin (0.1 microM) in the microdialysis perfusion buffer increased extracellular adenosine concentration in tumors to as high as 13 microM. These data show that extracellular adenosine levels in solid tumors are sufficient to suppress the local antitumor immune response and that interference with pathways of adenosine metabolism causes marked increases in tumor extracellular adenosine concentration. PMID- 9205064 TI - Tamoxifen interferes with the insulin-like growth factor I receptor (IGF-IR) signaling pathway in breast cancer cells. AB - The insulin-like growth factor I receptor (IGF-IR) is involved in the control of breast cancer cell growth. The cytostatic activity of tamoxifen (Tam), a nonsteroidal antiestrogen, is partially mediated through interference with IGF-I R-dependent proliferation, yet the effects of Tam on IGF-IR intracellular signaling have never been elucidated. Consequently, we investigated how Tam modifies the IGF-IR signaling pathway in estrogen receptor-positive MCF-7 breast cancer cells and in MCF-7-derived clones overexpressing either the IGF-IR (MCF 7/IGF-IR cells) or its major substrate, IRS-1 (MCF-7/IRS-1 cells). MCF-7/IGF-IR and MCF-7/IRS-1 cells exhibit greatly reduced estrogen growth requirements but retain estrogen receptors and express sensitivity to antiestrogens comparable to that in the parental cells. In all tested cell lines, regardless of the amplification of IGF signaling, a 4-day treatment with 10 nM Tam produced a similar cytostatic effect. In MCF-7 and MCF-7/IGF-IR cells, growth inhibition by Tam was associated with the reduced tyrosine phosphorylation of the IGF-IR in the presence of IGF-I; however, the basal level of the IGF-IR remained unaffected. Moreover, Tam inhibited both basal and IGF-I-induced tyrosine phosphorylation of IRS-1, which was accompanied by down-regulation of IRS-1-associated phosphatidylinositol 3'-kinase activity and reduced IRS-1/growth factor receptor bound protein 2 (GRB2) binding. In contrast, under the same treatment, tyrosine phosphorylation of Src-homology/collagen proteins (SHC; another substrate of the IGF-IR) and SHC/GRB2 binding were elevated. The protein levels of the IGF-IR and IRS-1 were not modified by Tam, whereas SHC protein expression was either not affected or moderately decreased by the treatment. In summary, this work provides the first evidence that in MCF-7 cells, cytostatic effects of Tam are associated with the modulation of IGF-IR signaling, specifically with: (a) down-regulation of IGF-I-induced tyrosine phosphorylation of the IGF-IR; (b) inhibition of IRS 1/phosphatidylinositol 3'-kinase signaling; and (c) up-regulation of SHC tyrosine phosphorylation and increased SHC/GRB2 binding. It is hypothesized that dephosphorylation of IRS-1 could be a major contributing factor in Tam cytostatic activity. PMID- 9205065 TI - A high ratio of insulin-like growth factor II/insulin-like growth factor binding protein 2 messenger RNA as a marker for anaplasia in meningiomas. AB - Insulin-like growth factors (IGFs) I and II have been implicated as autocrine or paracrine growth promoters. These growth factors bind to specific receptors, and the response is modulated by interaction with IGF-binding proteins (IGFBPs). We observed a strong correlation between anaplastic/atypical histopathology and a high IGF-II/IGFBP-2 mRNA ratio in a set of 68 sporadic meningiomas. A strong correlation was also found between clinical outcome and IGF-II/IGFBP-2 ratio, whereas previously used histochemical markers were less correlated to outcome. We suggest that a high IGF-II/IGFBP-2 mRNA ratio may be a sign of biologically aggressive behavior in meningiomas that can influence treatment strategies. We propose that low IGFBP-2 levels in combination with increased levels of IGF-II would result in more free IGF-II and consequently greater stimulation of proliferation. PMID- 9205066 TI - Cholera toxin triggers apoptosis in human lung cancer cell lines. AB - Cholera toxin (ChT) inhibits signals generated by multiple growth factors in human lung cancer cells, resulting in cell growth inhibition. We now report that ChT triggers apoptosis as shown by DNA fragmentation and activation of caspases cleaving poly(ADP-ribose) polymerase and lamin B. Apoptosis induced by ChT in a small cell lung cancer cell line is not affected by manipulations of intracellular cAMP through preincubation with isobutylmethylxanthine but can be modestly increased through inhibition of protein kinase C with chelerythrine. Thus, apoptosis is actively suppressed in lung cancer cells by a ChT-sensitive growth regulatory pathway, and these observations may have significant implications in the development of novel strategies for lung cancer treatment. PMID- 9205067 TI - p16INK4a promoter is hypermethylated at a high frequency in esophageal adenocarcinomas. AB - Loss of heterozygosity (LOH) of 9p21, which contains the p16INK4a tumor suppressor gene locus, is one of the most frequent genetic abnormalities in human neoplasia, including esophageal adenocarcinomas. Only a minority of Barrett's adenocarcinomas with 9p21 LOH have a somatic mutation in the remaining p16 allele, and none have been found to have homozygous deletions. To determine whether p16 promoter hypermethylation may be an alternative mechanism for p16 inactivation in esophageal adenocarcinomas, we examined the methylation status of the p16 promoter in flow-sorted aneuploid cell populations from 21 patients with premalignant Barrett's epithelium or esophageal adenocarcinoma. Using bisulfite modification, primer-extension preamplification, and methylation-specific PCR, we demonstrate that the methylation assay can be performed on 2 ng of DNA (approximately 275 cells). Eight of 21 patients (38%) had p16 promoter hypermethylation and 9p21 LOH, including 3 patients who had only premalignant Barrett's epithelium. Our data suggest that promoter hypermethylation with LOH is a common mechanism for inactivation of p16 in the pathogenesis of esophageal adenocarcinomas. PMID- 9205068 TI - Effects of tea, decaffeinated tea, and caffeine on UVB light-induced complete carcinogenesis in SKH-1 mice: demonstration of caffeine as a biologically important constituent of tea. AB - Oral administration of green or black tea inhibited UVB light-induced complete carcinogenesis in the skin of SKH-1 mice. Green tea was a more effective inhibitor than black tea. Oral administration of decaffeinated green or black tea resulted in substantially less inhibitory activity than did administration of the regular teas, and in one experiment, administration of a high-dose level of the decaffeinated teas enhanced the tumorigenic effect of UVB. Oral administration of caffeine alone had a substantial inhibitory effect on UVB-induced carcinogenesis, and adding caffeine to the decaffeinated teas restored the inhibitory effects of these teas on UVB-induced carcinogenesis. In additional studies, topical application of a green tea polyphenol fraction after each UVB application inhibited UVB-induced tumorigenesis. The results indicate that caffeine contributes in an important way to the inhibitory effects of green and black tea on UVB-induced complete carcinogenesis. PMID- 9205069 TI - Ornithine decarboxylase overexpression is a sufficient condition for tumor promotion in mouse skin. AB - In multistage tumorigenesis models, ornithine decarboxylase (ODC) is usually dysregulated at some point during tumor promotion, an early stage of carcinogenesis. To address the question whether constitutive overexpression of ODC would be a sufficient condition for tumor promotion, mice with high levels of ODC expression targeted to epidermal keratinocytes were used in skin tumorigenesis experiments. Transgenic mice with ODC targeted to hair follicle keratinocytes were much more sensitive than littermate controls to initiation with a single low dose of carcinogen; in fact, such mice no longer required treatment with tumor promoters for tumors to develop. Targeting ODC overexpression to both interfollicular and follicular keratinocytes did not further enhance tumor yield. Our results suggest that most, if not all, target cells for chemical carcinogens in the skin reside in hair follicles, and ODC overexpression is sufficient to activate such cells to expand clonally to form epidermal tumors. PMID- 9205070 TI - The putative tumor suppressor gene FHIT at 3p14.2 is rarely affected by loss of heterozygosity in primary human brain tumors. AB - To elucidate the role of the recently identified FHIT gene, located at 3p14.2 in human brain tumor carcinogenesis, a total of 259 tumors were analyzed for loss of heterozygosity (LOH) at microsatellite loci D3S1313, D3S1234, D3S1300, and D3S1481. In primary brain tumors, LOH was detected at a frequency of 8.4% (n = 214). Low-grade gliomas exhibited insignificantly lower LOH rates in comparison to high-grade gliomas (5.3%, n = 19, versus 11.1%, n = 90). Notably, no allelic loss was observed in 12 recurrent glioblastomas analyzed in comparison to their corresponding primary tumor lesions and in two astrocytomas with progression to higher grades of malignancy. Our data indicate that allelic loss of the FHIT gene is neither a critical event in carcinogenesis of primary brain tumors nor tumor grade-associated in astrocytic tumors. In contrast, observed LOH rate for brain metastases was as high as 54.5% (n = 45), in accordance with data thus far accumulated from analyses of corresponding primary tumors. PMID- 9205071 TI - Retinoic acid receptor alpha expression correlates with retinoid-induced growth inhibition of human breast cancer cells regardless of estrogen receptor status. AB - Retinoic acid receptor (RAR) alpha has been shown to play a role in retinoid induced growth inhibition of human breast cancer cell lines that express the estrogen receptor (ER). The dogma in the field has been that ER-positive breast cancer cell lines respond to retinoid treatment because they express RAR alpha, whereas ER-negative breast cancer cell lines are refractory to retinoid treatment and have been thought to express little or no RAR alpha. We set out to test several ER-negative breast cancer cell lines for expression of RAR alpha protein and responsiveness to retinoids in growth inhibition assays. Of six ER-negative breast cancer cell lines that were tested, one (SK-BR-3) had high levels of RAR alpha protein as measured by ligand-binding immunoprecipitation (approximately 55 fmol/mg protein) and also displayed sensitivity to growth inhibition by retinoids (9-cis-retinoic acid; EC50, approximately 3 nM). These cells were more sensitive than an ER-positive cell line, T-47D, which expressed approximately 35 fmol RAR alpha/mg total protein (9-cis retinoic acid; EC50, approximately 50-100 nM). Another ER-negative cell line, Hs578T, also expressed RAR alpha (approximately 23 fmol/mg) and was sensitive to retinoid-induced growth inhibition, albeit to a lesser extent than SK-BR-3 or T-47D cells. In contrast, the other ER-negative cell lines tested expressed low (<10 fmol/mg) or no detectable levels of RAR alpha protein and also did not respond to retinoids in growth inhibition assays. A RAR alpha agonist displayed 100 times greater potency than a RARgamma agonist in growth inhibition of both T-47D and SK-BR-3 cells, suggesting RAR alpha involvement in the process. Furthermore, a RAR alpha antagonist completely abolished the growth inhibition induced by RAR agonists, implying that the activity of the agonists is exerted solely through RAR alpha, not RARgamma, which is also expressed in both cell lines. Additionally, although retinoid X receptor (RXR) compounds are weakly active in growth inhibition of the RAR alpha-positive cell lines, they markedly increased the growth-inhibitory activity of RAR ligands. RXR compounds also potentiated the action of the antiestrogen 4 hydroxytamoxifen to inhibit the growth of T-47D cells. These findings have clinical ramifications in that patients with ER-negative tumors that are RAR alpha positive may be candidates for retinoid therapy. Additionally, combinations of RXR ligands with RAR ligands (especially RAR alpha agonists) and/or antiestrogens may have utility in the treatment of breast cancer. PMID- 9205072 TI - Expression of human prostate-specific glandular kallikrein protein (hK2) in the breast cancer cell line T47-D. AB - Human glandular kallikrein (hK2) protein, like prostate-specific antigen (PSA), is produced mainly in prostatic epithelium. It may be useful as a new diagnostic indicator for prostate cancer. Recently, a number of hK2-specific monoclonal antibodies have been developed that enable us to detect hK2 protein in human prostate tissue, seminal fluid, and sera. Whether hK2 can be expressed, like PSA, in nonprostatic cells is not known. In this study, we have characterized the presence of hK2 in an androgen-responsive breast cancer cell line T47-D at both the protein and mRNA levels with an immunoassay, Western blot analysis, Northern blot analysis, and the reverse transcription-PCR. Using a sensitive immunoassay with monoclonal antibodies to hK2, we found that T47-D cells could be induced with androgens, mineralocorticoids, glucocorticoids, and progestins to produce significantly more hK2 than PSA. Estrogens failed to mimic the effect of the other steroids, blocking instead the stimulatory effect of androgens. Androgen induction of hK2 in T47-D cells was dose dependent. More interestingly, we found that the hK2 in androgen-induced T47-D cell spent media appears to be the pro form of hK2 rather than mature hK2. Our study demonstrates that hK2, a serine protease thought to be found only in prostate-related tissues and fluids, is also produced in a breast cancer cell line T47-D after steroid stimulation. This finding suggests that hK2 may have a potential role in breast cancer as well as prostatic cancer and will be the impetus for further studies of hK2 distribution and function. PMID- 9205073 TI - Synergy between tamoxifen and cisplatin in human melanoma cells is dependent on the presence of antiestrogen-binding sites. AB - We have demonstrated previously that cisplatin (DDP) and tamoxifen (TAM) act synergistically to kill human melanoma T-289 cells, and that the observed synergy is lost in the 3-fold TAM-resistant subline, 289/TAM6. We have identified the intracellular antiestrogen-binding sites (AEBSs), defined by their ability to bind antiestrogens while having no affinity for estrogen, as a possible mediator of this synergy. We report here that [3H]TAM binds to AEBSs, as defined by the ability of N,N-diethyl-2-[4-(phenylmethyl)phenoxy]ethanamine-HCl, an AEBS specific ligand, to compete with [3H]TAM binding. Furthermore, we have characterized the number of binding sites and their affinity for [3H]TAM by Scatchard analysis in whole-cell lysates, microsomal fractions, and nuclear fractions of both cell lines by competing [3H]TAM binding with increasing concentrations of unlabeled TAM. These data demonstrate that the loss of a high affinity AEBS from the nuclear fraction of the 289/TAM6 cell line correlates with the loss of synergy between DDP and TAM in these cells. This implicates AEBSs as a critical component of the mechanism that mediates the synergistic interaction of DDP and TAM in human melanoma cells. PMID- 9205074 TI - HSP27 as a mediator of confluence-dependent resistance to cell death induced by anticancer drugs. AB - Resistance of colorectal cancer cells to chemotherapeutic drugs increases as cells reach confluence. Here we show that the small stress protein HSP27, which has been described to block necrotic and apoptotic cell death, accumulates in confluent human colorectal cancer cell lines HT-29 and Caco2. Cell confluence also induces HSP27 phosphorylation and changes in its intracellular distribution. We also show that overexpression of human HSP27 by transfection of HT-29 cells increased the resistance of cells to doxorubicin or cisplatin and prevented drug induced apoptosis. Interestingly, nonconfluent HSP27-transfected cells and confluent control cells in which HSP27 is expressed at the same level displayed a similar drug resistance. HSP27-transfected cells did not exhibit an enhanced resistance when they reached confluence, nor was there an increased accumulation of HSP27. We have previously shown that HSP27 expression blocks tumor necrosis factor-induced cell death as a result of decreasing intracellular reactive oxygen species (ROS). Here we show that HSP27 overexpression in HT-29 cells, obtained either by transfection or by growing the cells at high density, correlated with a significant ROS decrease. We conclude that cell confluent-dependent HSP27 accumulation, probably due to its ability to decrease ROS levels, is essential for the establishment of the resistance of colorectal cancer cells when reaching confluence. PMID- 9205076 TI - Ifosfamide cytotoxicity on human tumor and renal cells: role of chloroacetaldehyde in comparison to 4-hydroxyifosfamide. AB - The initial metabolism of ifosfamide (IFO) consists of two different pathways, which lead to the alkylating metabolite 4-hydroxy-IFO and to chloroacetaldehyde (CAA). CAA has been reported to cause side effects, such as neuro- and nephrotoxicity, whereas no direct antitumor effect has been described thus far. Therefore, two human tumor cell lines (MXI and S117) and a renal tubular cell line (Landa Leiden) were exposed to 4-hydroxy-IFO, CAA, and a combination of both. The concentrations used were in the same range as measured in the blood of 10 patients treated with 5 g/m2 IFO. The cell survival was measured using the MTT assay. Similar dose-response curves were found for both metabolites. For the MX1 tumor, the IC50s of 4-hydroxy-IFO and CAA were 10.8 and 8.6 microM, respectively. For the reduction of S117 cell survival, higher concentrations of the metabolites were needed (25.0 microM 4-hydroxy-IFO and 15.3 microM CAA). Combination treatment of the cells resulted in an approximately additive effect. Both metabolites exhibited similar toxicity against Landa Leiden cells. Our results indicate that CAA has its own cytotoxic profile against tumor cells. Hence, we conclude that the molecular mechanism of action of IFO seems to be only in part an alkylating effect and that CAA may play an important role in the therapeutic efficacy of IFO. PMID- 9205075 TI - Vitamin E succinate inhibits proliferation of BT-20 human breast cancer cells: increased binding of cyclin A negatively regulates E2F transactivation activity. AB - Vitamin E succinate (VES) inhibited the proliferation of the estrogen receptor negative human breast cancer cell line, BT-20, in the G1 phase of the cell cycle. The E2F proteins are integral transcriptional components in the regulation of cell growth. Overexpression of E2F-1 blocked the ability of VES to inhibit BT-20 cell growth, suggesting that VES regulation of E2F-1 activity leads to growth arrest of BT-20 cells. VES, although having little effect on E2F-1 steady-state protein levels, decreased E2F-1 phosphorylation and transactivation activity and increased cyclin A binding to E2F-1. GAL4-E2F-1 deletion mutant studies indicated that cyclin A negatively regulates E2F function. In VES-treated BT-20 cells, the cyclin A protein exhibited reduced kinase activity, which correlated with decreased steady-state levels and binding of cyclin-dependent kinase-2 to cyclin A and increased steady-state levels and binding of p21cip1 to cyclin A and cyclin dependent kinase-2. The functional consequence of the negative regulatory effect of VES on E2F-1 function was shown by the ability of VES to inhibit the transcriptional activation of an E2F-1 responsive gene, c-myc. These studies show that VES induces growth inhibition of BT-20 cells through a mechanism that involves cyclin A-negative regulation of E2F-mediated transcription. PMID- 9205077 TI - Intratumoral pharmacokinetics and in vivo gene expression of naked plasmid DNA and its cationic liposome complexes after direct gene transfer. AB - The pharmacokinetic properties and gene expression of naked plasmid DNA and its cationic liposome complexes were studied after direct intratumoral injection. Using a Walker 256 tissue-isolated tumor perfusion system, we quantified the recovery of naked plasmid DNA and cationic liposome complexes in the tumor, leakage from the tumor surface, and the venous outflow after intratumoral injection. Approximately 50% of naked plasmid DNA had been eliminated from the tumor 2 h after injection, whereas more than 90% of plasmid DNA was retained in the tumor when it was complexed with cationic liposomes. However, the distribution of these complexes in the tumor was restricted to the tissue surrounding the injection site. Pharmacokinetic analysis of the venous outflow profiles suggested that the rate-limiting process that determines the retention of plasmid DNA in the tumor is transferred from the injection site in the tumor tissue and that complexation with cationic liposomes may retard this process. Furthermore, we examined the gene expression of chloramphenicol acetyltransferase DNA constructs (naked pCMV-CAT) and the corresponding cationic liposome [3-beta (N-(N',N'-dimethylaminoethane)carbamoyl)cholesterol] complexes. A similar level of gene expression was observed in vivo after direct intratumoral injection of naked DNA and its cationic liposome complexes. In both cases, a great variation was observed between tumors, and localization of gene-transduced cells in the tumor tissue was limited to the area in the vicinity of the injection site. Thus, these pharmacokinetic and gene expression studies have demonstrated that cationic liposomes can enhance the retention of injected DNA in the tumor model, whereas cationic liposome complex does not necessarily improve gene expression because of its poor dissemination in this tumor. The present study also suggested that there is a need to control the behavior of the injected naked plasmid DNA and its cationic liposome complexes to ensure better distribution throughout the tumor. PMID- 9205078 TI - Insulin-like growth factor 1 (IGF-1) alters drug sensitivity of HBL100 human breast cancer cells by inhibition of apoptosis induced by diverse anticancer drugs. AB - In this study, we tested the hypothesis that insulin-like growth factor-1 (IGF-1) modulates apoptosis in human breast cancer cells, HBL100, induced by diverse chemotherapeutic drugs. IGF-1 increased cell survival of HBL100 cells treated with 5-fluorouracil (antimetabolite), methotrexate (antimetabolite), tamoxifen (antiestrogen/antiproliferative), or camptothecin (topoisomerase 1 inhibitor) and after serum withdrawal. Elevated cell survival was not due to an increase in cell proliferation by IGF-1, but rather to an inhibition of apoptosis. Evidence for death by apoptosis was supported by cellular morphology and DNA fragmentation. There were no changes observed in Bcl-2 protein or bax mRNA levels. Extracellular matrix (ECM) is known to influence the apoptotic response of cells; therefore, the antiapoptotic effect of IGF-1 on breast cancer cells was examined using different ECMs: laminin, collagen IV, or Matrigel. IGF-1 protected cells from apoptosis induced by methotrexate on all ECMs tested, providing the first evidence that IGF-1 protects against apoptosis in three-dimensional culture systems. These data provide the rationale to search for drugs that lower serum IGF-1 in an effort to improve the efficacy of chemotherapeutic drugs for the treatment of breast cancer. PMID- 9205079 TI - Apoptosis in the subependyma of young adult rats after single and fractionated doses of X-rays. AB - Ionizing radiation is commonly used in the treatment of brain tumors but can cause significant damage to surrounding normal brain. The pathogenesis of this damage is uncertain, and understanding the response of potential target cell populations may provide information useful for developing strategies to optimize therapeutic irradiation. In the mammalian forebrain, the subependyma is a mitotically active area that is a source of oligodendrocytes and astrocytes, and it has been hypothesized that depletion of cells from this region could play a role in radiation-induced white matter injury. Using a distinct morphological pattern of nuclear fragmentation and an immunohistochemical method to specifically label the 3'-hydroxyl termini of DNA strand breaks (terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling), we quantified apoptosis in the subependyma in the young adult rat brain after single and fractionated doses of X-rays. Significant increases in apoptotic index (percentage of cells showing apoptosis) were detected 3 h after irradiation, and the peak apoptotic index was detected at 6 h. Six h after irradiation, the dose response for apoptosis was characterized by a steep increase in apoptotic index between 0.5 and 2.0 Gy and a plateau from 2-30 Gy. The fraction of cells susceptible to apoptosis was estimated to be about 40%, and treatment of rats with cycloheximide inhibited apoptosis. When daily 1.5-Gy fractions of X-rays were administered, the first three fractions were equally effective at decreasing the cell population via apoptosis. There was no additional apoptosis or decrease in cellularity in spite of one to four additional doses of X-rays. Those data suggested some input of cells into the subependymal population during fractionated treatment, and subsequent studies showed that there was a significant rise in 5-bromo-2' deoxyuridine labeling index 2-3 days after irradiation, indicating increased cellular proliferation. The proliferative response after depletion of cells via apoptosis may represent the recruitment of a relatively quiescent stem cell population. It is possible that the radiation response of subependymal stem cells and not the apoptotic-sensitive population per se are critical elements in the response of the brain to radiation injury. PMID- 9205080 TI - Chromosome 9p deletions in invasive and noninvasive nonfunctional pituitary adenomas: the deleted region involves markers outside of the MTS1 and MTS2 genes. AB - We have screened 57 cases of primary, nonfunctional, pituitary adenomas for loss of heterozygosity of markers on chromosome 9p. Using a panel of 11 microsatellite markers, we found hemizygous deletion with at least one of the markers in 18 tumors (31.5%). The frequency of loss was similar in both noninvasive (8 of 26; 31%) and invasive tumors (10 of 31; 32%), suggesting that loss on this chromosome might be an early event in pituitary tumorigenesis. Two discrete areas of loss were punctuated by a region of retention of heterozygosity between the markers D9S171 and IFNA, indicative of homozygous deletion. However, multiplex PCR analysis (MTS1 and MTS2) and the presence of a 3' untranslated region polymorphism in MTS1 suggested that neither of these tumor suppressor genes was homozygously deleted. In 6 of the 18 tumors showing LOH, sufficient DNA was also available for Southern blot analysis and, in all cases, showed retention of MTS1. Cell mixing experiments of tumor cell DNA homozygously deleted for MTS1 with DNA in which neither copy of the gene was deleted only gave rise to a signal at contamination levels greater than 30% and could discriminate homozygous and hemizygous loss. These studies support the recent findings that mechanisms other than hemi- and homozygous deletion are most likely responsible for the loss of MTS1 gene product in pituitary tumors (M. Woloschak et al., Cancer Res., 56: 2493 2486, 1996.). These data show that losses on either side of 9p21-22, both or either of which may be deleted, are involved in pituitary tumorigenesis and provide evidence for distinct suppressor gene loci, in addition to MTS1, on chromosome 9p. PMID- 9205081 TI - Genetic alterations of p53 and ras genes in 1,3-butadiene- and 2',3' dideoxycytidine-induced lymphomas. AB - Mutations of p53 and ras genes were analyzed in 40 and 31 1,3-butadiene (BD) induced lymphomas of B6C3F1 mice (BLFs), respectively, and in 63 2',3' dideoxycytidine-induced lymphomas, which were collected from B6C3F1 (n = 16) or NIH Swiss mice (DLSs; n = 47). The frequencies of K- and N-ras mutations in BLFs (32 and 13%, respectively) were higher than those in DLSs (13 and 2%, respectively). Seven of 10 K-ras-mutated BLFs contained codon 13 CGC mutations, whereas no mutation in K-ras codon 13 was detected in DLSs, suggesting that the codon 13 CGC mutation is specific for BD exposure. Interestingly, 8 of 13 BLFs with ras mutations were from low-dose (< or = 200 ppm) or stop-exposure (26 weeks) groups. These results suggest that ras mutations play an important role in the development of BD-induced lymphoma and may represent an early event. Analysis of genetic alterations in exons 5-8 of the p53 gene revealed mutations in seven of the BLFs and three of the DLSs. All seven BLFs carrying p53 mutations were collected from the high-dose (625 ppm) continuous exposure group, which might indicate that p53 is involved in the progression of BD-induced lymphoma and in late stage of lymphomagenesis. Mutations in ras and p53 genes are relatively infrequent in 2',3'-dideoxycytidine-induced lymphomas, suggesting that other genes must be involved. PMID- 9205082 TI - A mutation in the MSH5 gene results in alkylation tolerance. AB - DNA methylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) are potent carcinogens; their carcinogenic effect is mainly due to the effect of production of O6-methylguanine (O6 MeG) on DNA. O6 MeG is not only mutagenic but also toxic to the cell because Mer-/Mex- cells unable to remove O6 MeG are very sensitive to killing by MNNG. It has been proposed that repeated futile mismatch correction of O6 MeG-containing bp is responsible for the genotoxicity of the O6 MeG lesion and that loss of mismatch repair activity results in cellular tolerance to O6 MeG, but the hypothesis has not been proved. We used yeast as a model to test this hypothesis and found that chromosome deletion of any known nuclear mitotic mismatch repair genes, including MLH1, MSH2, MSH3, MSH6, and PMS1, did not rescue mgt1delta O6 MeG DNA repair methyltransferase-deficient cells from killing by MNNG. A large number of mgt1delta, MNNG-tolerant revertants were isolated, among which one cell line, XS-14, has been found to carry a mutated allele of the MSH5 gene. The mutation also affected spore survival but did not increase the spontaneous mutation rate. We further demonstrated that a mutated form of the MSH5 gene, msh5-14, not the msh5delta-null mutation, is responsible for the cellular tolerance to MNNG in XS-14 cells. This observation offers an alternative model that may reconcile seemingly contradictory observations of yeast and mammalian cells. PMID- 9205083 TI - A general role for c-Fos in cellular protection against DNA-damaging carcinogens and cytostatic drugs. AB - One of the earliest responses of cells upon exposure to DNA-damaging agents is the induction of c-fos. To elucidate the biological role of Fos expression, we analyzed cells deficient in c-Fos upon treatment with different DNA-damaging agents, including carcinogens and antineoplastic drugs. We show that cells lacking c-Fos are hypersensitive with regard to reproductive cell death, apoptosis, and chromosomal breakage after treatment with agents inducing methylation lesions, bulky adducts, or crosslinks in DNA. They were not significantly hypersensitive to ionizing radiation. The activities of various repair enzymes and glutathione S-transferase and the level of proliferating cell nuclear antigen were not altered in c-fos-/- fibroblasts. Furthermore, the cells were able to remove the main methylation lesions from DNA. c-Fos-deficient cells exhibited a more severe mutagen-induced block to DNA replication and were compromised in the abolition of replication blockage. The data provide compelling evidence that c-Fos/activator protein-1 plays a decisive and general role in cellular defense against genotoxic agents, which require DNA replication to induce chromosomal instability. They are consistent with the hypothesis that impaired recovery from DNA replication inhibition upon mutagen exposure is causally involved in c-fos-/- hypersensitivity. PMID- 9205084 TI - Ectopic expression of 2-5A synthetase in myeloid cells induces growth arrest and facilitates the appearance of a myeloid differentiation marker. AB - Two variants of the human myeloid leukemia cell line HL-60 were used to study the possible involvement of the IFN-induced protein 2-5A synthetase in cell growth arrest and differentiation. The two variants, HL-205 and HL-525, are equally susceptible to differentiation to the granulocyte lineage by exposure to DMSO, but only HL-205 cells acquire the macrophage phenotype following exposure to phorbol esters. The kinetics of 2-5A synthetase activity was established in both variants exposed to either DMSO or phorbol 12-myristate 13-acetate. With DMSO treatment, 2-5A synthetase activity was markedly induced in both variants, although with slightly different kinetics. With phorbol 12-myristate 13-acetate treatment, 2-5A enzymatic activity increased only in HL-205; no activity was detected up to 96 h after treatment in HL-525. The induction of 2-5A synthetase activity is apparently alpha/beta-IFN dependent, because only antibodies directed against a mixture of alpha- and beta-IFN completely abolished the increase in activity detected during differentiation of HL-205 cells. To directly establish the role of 2-5A synthetase in differentiation, HL-205 cells were transfected with an expression vector harboring the cDNA for the 43-kDa isoform of murine 2 5A synthetase fused to the inducible metallothionein promoter. Two clones, clone 6, which yielded a low level of 2-5A synthetase activity in response to ZnCl2 (which activates the promoter), and clone 7, which was a high responder, were further analyzed and compared with the control clone, neo. Reductions in the rates of cell growth and thymidine incorporation were observed with both clone 6 and clone 7 cells exposed to ZnCl2; clone 7 was more responsive. In addition, the level of c-myc-specific RNA transcript was greatly reduced in ZnCl2 or beta-IFN treated clone 7 cells, whereas the neo cells responded similarly only after beta IFN treatment. Treatment of clone-neo cells with beta-IFN resulted in conversion of pRb protein from the phosphorylated to the underphosphorylated form within 24 h; ZnCl2 had no effect, even after 72 h. In contrast, the accumulation of the underphosphorylated form of pRb was observed in clone 7 cells treated either with beta-IFN or ZnCl2. Finally, a significant increase in nitro blue tetrazolium positive cells, an indication of differentiation, was evident with ZnCl2-treated clone 6 and clone 7 cells; no such increase was observed with clone-neo cells under similar conditions. We conclude that ectopic expression of 2-5A synthetase in HL-205 cells results in cell growth arrest and facilitates the appearance of a myeloid differentiation marker. PMID- 9205085 TI - Cadherin-6, a cell adhesion molecule specifically expressed in the proximal renal tubule and renal cell carcinoma. AB - Cadherins are a family of calcium-dependent, cell-cell adhesion molecules that play an important morphoregulatory role in a wide variety of tissues. Alterations in cadherin function have been implicated in tumor progression in a number of adenocarcinomas. Despite the increasing number of new cadherins identified, little is known about cadherins in normal renal tissue and renal carcinomas. A novel cadherin transcript, cadherin-6, was recently described to be present in renal cancer cell lines and fetal kidney, but no data on protein expression nor tissue localization has been reported. In this study, we demonstrate that the expression of cadherin-6 is restricted to the proximal tubule epithelium. This finding is critical because these cells give rise to the majority of neoplasms of this organ. Furthermore we demonstrate typical cadherin features of cadherin-6, including cytoplasmic binding to alpha- and beta-catenin. We present data of cadherin-6 expression in a series of 32 primary renal cell cancers. Cadherin-6 expression tended to vary with histology in these samples. Whereas the majority of renal cell cancers with histology-associated poor prognosis (i.e., high grade clear cell carcinomas and sarcomatoid renal tumors) show aberrant expression of cadherin-6, in tumors with a favorable prognosis (i.e., low grade clear cell carcinomas and papillary cancers), normal cadherin-6 expression was predominant. Overall, these findings demonstrate specific expression of cadherin-6 in the proximal renal tubules in normal human kidney and suggest that alterations of cadherin-6 expression are associated with progression of renal cell carcinoma. PMID- 9205086 TI - Glutathione S-transferase expression in hepatitis B virus-associated human hepatocellular carcinogenesis. AB - Hepatitis B virus (HBV) and aflatoxin B1 represent the main risk factors for the development of hepatocellular carcinoma (HCC) in areas endemic for liver cancer. The glutathione S-transferases (GSTs) are a family of Phase II detoxification enzymes that catalyze the conjugation of a wide variety of endogenous and exogenous toxins, including aflatoxin B1, with glutathione. This study characterizes the GST isoenzyme composition (alpha, mu, and pi) of both HBV infected normal hepatic tissues and HCCs. Analysis of matched pairs of hepatic tissue (normal and tumor) from 32 HCC patients indicated that total GST activity was significantly higher in normal tissues than in tumor tissues, although the percentage of samples expressing GST alpha and pi was equivalent. GST mu was detected by Western blot in the normal tissue from 87.5% of the subjects possessing the GST M1 gene but only 28.6% of the corresponding tumor tissues. The GST activity of normal tissue from GST M1 null patients was significantly decreased as compared to that of subjects possessing the GST M1 gene (264.6 and 422.2 nmol/min/mg, respectively; P = 0.005). GST pi appeared to be overexpressed in the normal tissue of GST M1 null patients, a potential compensatory effect. Patients positive for HBV DNA had significantly lower GST activity than those who were HBV negative (302.1 versus 450.0 nmol/min/mg, respectively; P = 0.02). These results suggest that cellular protection within the human liver is compromised by HBV infection and further decreased during hepatocellular tumorigenesis. PMID- 9205087 TI - Biodistribution of 125I-labeled des(1-3) insulin-like growth factor I in tumor bearing nude mice and its in vitro catabolism. AB - Insulin-like growth factor I (IGF-I) is a potent mitogen for many tumor cell lines, and IGF-I receptors are overexpressed in many tumors. Specific IGF-binding proteins (IGFBPs) modulate the interaction of IGF and its receptors. Consequently, radiolabeled IGF-I has been considered for tumor imaging. In the present study, we investigated the biodistribution of 125I-labeled des(1-3)IGF-I, a truncated analogue of IGF-I, in tumor-bearing nude mice. Additional studies included its catabolism by tumor cells in vitro and its binding to serum IGFBPs in vivo in nude mice. We also compared groups that were and were not injected with unlabeled peptide analogue. Our data showed that 125I-labeled des(1-3)IGF-I catabolized very fast, with a rapid appearance of nonprecipitable iodine, when incubated at 37 degrees C, but it was not catabolized at 4 degrees C incubation. 125I-labeled des(1-3)IGF-I was bound to serum-binding proteins, mainly in a complex with a molecular weight of M(r) 150,000. The uptake of radioactivity in normal tissues decreased quickly with time, particularly in the kidneys. In mice receiving higher doses of des(1-3)IGF-I, the radioactivity in all normal tissues was lower than in the mice with no carrier-added des(1-3)IGF-I, except in the stomach and spleen. These data suggest that 125I-labeled des(1-3)IGF-I is rapidly internalized after binding to the IGF receptor and is rapidly catabolized with release of breakdown products. Lower specific activity of 125I-labeled des(1 3)IGF-I resulted in altered biodistribution, including faster blood clearance and higher tumor uptake, by decreasing the formation of complexes with IGFBPs. PMID- 9205088 TI - Telomerase activity in benign endometrium and endometrial carcinoma. AB - Telomerase, a ribonucleoprotein associated with synthesis of telomeric DNA, is postulated to play a role in cellular senescence and immortalization. Telomerase adds a hexonucleotide telomeric sequence to the chromosomal ends during replication and is preferentially expressed in most malignant and germ-line tissues but is usually undetectable in normal somatic cells. In the current study, 34 human endometrial tissues (20 malignant and 14 benign) were analyzed for telomerase activity by a nonradioactive PCR-based method using the TRAP-eze telomeric repeat amplification detection kit (Oncor). Nineteen of 20 (95%) endometrial carcinomas and 8 of 8 (100%) benign endometrial tissues from premenopausal women exhibited strong telomerase activity, whereas 6 of 6 (100%) benign endometrial tissues from postmenopausal women showed only weak telomerase activity. There was no correlation of telomerase activity with tumor grade, depth of invasion, or DNA content. Benign cycling endometrium, a rapidly proliferating tissue, features positive telomerase activity, although expression in nonneoplastic tissues has only rarely been previously reported. Only weak activity is detected in endometrial tissues after menopause, but telomerase activity can be strongly reactivated in patients who develop endometrial cancer. PMID- 9205089 TI - Chromosomal imbalance maps of malignant solid tumors: a cytogenetic survey of 3185 neoplasms. AB - To assess the distribution of gains and losses of genetic material in malignant solid neoplasms, 11 tumor types for which at least 100 short-term cultured cases with clonal chromosome aberrations had been reported in the literature were selected. The study was based on cytogenetic information from 508 breast carcinomas, 447 malignant neuroglial tumors, 435 kidney carcinomas, 333 colon carcinomas, 304 ovarian carcinomas, 303 lung carcinomas, 209 testicular germ cell tumors, 206 head and neck carcinomas, 172 malignant melanomas, 142 Wilms' tumors, and 126 neuroblastomas. In each case, the net imbalances were calculated for each chromosome band. The profiles of gains and losses revealed that all tumor types display unique combinations of imbalances. However, there is also considerable overlap among the profiles of the different diagnostic entities, indicating that similar molecular mechanisms may be operative in the development of many types of neoplasia. Deletions were more common than gains in all tumor types, with chromosomes X, Y, 4, 10, 13-15, 18, and 22 and chromosome segments 1p22-pter, 3p13-pter, 6q14-qter, 8p, 9p, and 11p being particularly often deleted in the majority of tumors. To better delineate critical lost segments, deletion profiles based only on structural rearrangements were made for chromosomes 1, 3-12, and 17, which all had on average at least four registered deletions per band. The relative distribution of losses indicated that different bands/regions are affected in different tumor types and that, often, several distinct candidate tumor suppressor gene loci can be discerned within the same chromosome arm, e.g., 1p12-13, 1p22, 1p34, and 1p36 on the short arm of chromosome 1 and 7q22, 7q32, and 7q36 on the long arm of chromosome 7. The only chromosomes or chromosome segments more often gained than deleted were chromosomes 7 and 20 and the long arms of chromosomes 1 and 12, suggesting the presence there of dominantly acting growth-regulatory genes. The data presented in this study should be valuable as a guide for molecular genetic studies on allelic imbalances and for the interpretation of results from studies using comparative genomic hybridization. PMID- 9205090 TI - Optimal ratios of biliary glycoprotein isoforms required for inhibition of colonic tumor cell growth. AB - Rodent biliary glycoprotein (Bgp), also known as C-CAM, has recently been shown to function as a tumor suppressor in colon, prostate, and bladder cancers. This glycoprotein is a member of the carcinoembryonic antigen family and is one of the only proteins in this family to encode either a long (71-73 amino acids) or short (10 amino acids) cytoplasmic domain. We and others have shown that the growth inhibitory properties of Bgp depend upon the expression of its long cytoplasmic domain. However, the two Bgp isoforms normally coexist in most cell types surveyed; the longer variant is usually present in lower amounts than the shorter one. In this study, we have examined the in vitro and in vivo growth properties of both mouse Bgp variants separately and in combination. To determine the physiologically relevant expression levels and ratios of the two Bgp variants, we have quantified the amount of the longer variant in normal colonic epithelial cells and showed that it constitutes 15-20% of total Bgp expressed in this tissue. To mimic the in vivo situation, we have generated double transfectant cell lines expressing the longer and shorter Bgp isoforms coordinately in tumorigenic CT51 mouse colonic carcinoma cells and demonstrated that the longer Bgp isoform exhibits a dominant tumor growth inhibition phenotype over that of the shorter variant within physiological levels of expression of Bgp. Unexpectedly, significant overexpression of the longer Bgp isoform alone led to reversal of the tumor inhibition phenotype. These results, therefore, suggest that there may be a limiting threshold of Bgp expression or Bgp-associating proteins mediating the tumor inhibition phenotype. PMID- 9205092 TI - Increased expression of low density lipoprotein receptor-related protein/alpha2 macroglobulin receptor in human malignant astrocytomas. AB - Low-density lipoprotein receptor-related protein (LRP) plays an important role in regulating proteinase activity, which is necessary for cellular invasive processes. In this study, we investigated the presence of both LRP and urokinase type plasminogen activator receptor (uPAR) in astrocytoma tissues and in glioma cell lines by PCR and immunohistochemical analysis. LRP mRNA was expressed frequently in glioblastomas, as compared with low-grade astrocytomas by PCR analysis and was well correlated with uPAR expression. These results were consistent with the immunohistochemical localization of LRP in glioblastomas. Immunohistochemistry of LRP on sequential frozen sections showed that neoplastic glial cells and endothelial cells of glioblastomas exhibited intense LRP immunoreactivity, whereas LRP was almost undetectable in low-grade astrocytomas and in normal glial cells and endothelial cells of normal brain tissues. In normal brain tissues, LRP immunoreactivity was identified in the pyramidal neurons of the cerebral cortex. In metastatic brain tumors (metastatic lung adenocarcinomas) and primary lung adenocarcinomas, LRP expression was low to undetectable, suggesting that LRP expression is regulated differently in these tumors than in malignant astrocytomas. These results indicate that LRP is overexpressed in malignant astrocytomas, especially in glioblastomas, and the increased expression of LRP appears to correlate with the expression of uPAR and the malignancy of astrocytomas. Our results suggest strongly that LRP may play a role in facilitating glioblastoma invasiveness and neovascularization within tumor tissues by regulating cell surface proteolytic activity. PMID- 9205093 TI - Correspondence re: C.V. Rao, et al., Inhibition of 2-amino-1-methyl-6 phenylimidazo[4,5-b]pyridine-induced lymphoma formation by oltipraz. Cancer Res., 56: 3395-3398, 1996. PMID- 9205091 TI - Two cytodifferentiation agent-induced pathways, differentiation and apoptosis, are distinguished by the expression of human papillomavirus 16 E7 in human bladder carcinoma cells. AB - Many transformed cells have been found to lose the capacity to proliferate and undergo differentiation following exposure to hybrid polar agents. This study investigates the mechanism by which hexamethylene bisacetamide (HMBA) suppresses the proliferation of the human bladder carcinoma line, T24. We found that following a 24-h exposure to HMBA, T24 proliferation was inhibited, and cells arrested in G1 phase and underwent morphological maturation. HMBA-induced cessation of proliferation was mediated, in part, by effects on cell cycle regulatory proteins. In T24 cells cultured without HMBA, E2F complexes predominantly with p107. In culture with inducer, p107 protein decreased, pRB and p130 were converted to underphosphorylated forms, and E2F was shifted into complexes with pRB and p130. To determine whether the formation of pRB:E2F and p130:E2F complexes was required for the HMBA-induced G1 arrest, the ability of the pocket proteins to bind E2F was blocked by enforced expression of human papillomavirus 16 E7. Following culture with HMBA, the T24 clones expressing E7 died, whereas vector-alone T24 clones arrested in G1 phase. T24/E7-1 cells did not form pRB:E2F or p130:E2F complexes upon culture with HMBA; rather, E2F was present in its free form. T24/E7-1 cells cultured with HMBA initially accumulate in G1. By day 2, they have entered into S phase, and by day 3, over 80% of the cells became apoptotic. Taken together, these studies enlarge the repertoire of demonstrated developmental pathways that may be triggered in transformed cells, depending upon their molecular status, and may provide potential therapeutic opportunities for cancer. PMID- 9205094 TI - Relationship of p215BRCA1 to tyrosine kinase signaling pathways and the cell cycle in normal and transformed cells. AB - We have analysed the relationship of the products of two genes, neu and BRCA1, known to be important in human breast cancer. Highly specific antibodies that recognized both the rodent and human form of the BRCA1 gene product (Mr 215 kDa, p215BRCA1) were developed to facilitate these efforts. p215BRCA1 was identified as a tyrosine phosphorylated protein primarily localized in the nucleus of several breast cancer cell lines. In transformed murine and human cells, levels of p215BRCA1 tyrosine phosphorylation were inversely correlated with the activity of the erbB family receptor-tyrosine-kinases and with the transformed growth features of these cells. Regulation of p215BRCA1 tyrosine phosphorylation was also related to events in the cell cycle. Increased levels of p215BRCA1 phosphotyrosine content were observed in NIH3T3 cells arrested at the G2/M transition. These findings indicate that the products of BRCA1, neu, and erbB breast cancer genes participate in a common or shared signaling pathway important in cell growth and its regulation. PMID- 9205095 TI - p94fer facilitates cellular recovery of gamma irradiated pre-T cells. AB - p94fer is a ubiquitous, nuclear and cytoplasmic tyrosine kinase, whose accumulation has been demonstrated in all mammalian cell lines analysed. In the present work, the p94fer expression profile was determined in cell lines which were not tested before. While being present in several hematopoietic and non hematopoietic cell lines including thymic stromal cells, the p94fer kinase could not be detected in pre-T and T cell lines. p94fer was also absent in pre-B line, but accumulated in these cells upon their induced development to antibody producing cells. This is in agreement with the absence of p94fer in primary thymic and splenic T lymphocytes and its induced accumulation in stimulated B cells. Relatively high p94fer levels were detected in primary thymic and splenic stromal cells. Ectopic expression of p94fer in pre-T cells slightly affected their cell cycle profile but it did not affect their apoptotic death which was induced by ionizing radiation. However, p94fer facilitated dramatically, the cellular recovery of gamma irradiated pre-T cells which have escaped the apoptotic death. The enhanced recovery of the irradiated, p94fer expressing pre-T cells, resulted most probably from their increased survival, rather than from a prominent change in their proliferation rate. The absence of p94fer from pre-B and pre-T cells, may thus contribute to the relative sensitivity of these cells to ionizing radiation and to their dependence on the functioning of other nuclear tyrosine kinasese. PMID- 9205096 TI - Differences in kappaB DNA-binding properties of v-Rel and c-Rel are the result of oncogenic mutations in three distinct functional regions of the Rel protein. AB - The oncogene v-rel of Reticuloendotheliosis virus, strain T, is derived from an avian c-rel proto-oncogene. c-rel encodes a member of the Rel/NF-kappaB family of transcription factors. The highly oncogenic v-Rel differs from c-Rel which has low transforming potential by the acquisition of numerous mutations. In this manuscript, we demonstrate that the oncogenic mutations in v-Rel directly alter the ability of this protein to bind to DNA. Electrophoretic mobility shift analysis with Rel proteins synthesized in vitro as well as isolated from nuclei of Rel expressing cells showed that three mutation clusters, present in the N terminus, the center and the C-terminus of v-Rel, altered three different aspects of DNA binding. In contrast, the oncogenic C-terminal deletion of 118 amino acids present in v-Rel had almost no influence on its DNA binding. The N-terminal mutation cluster altered the kappaB DNA-binding specificity of the v-Rel oncoprotein relative to c-Rel. The mutation Met-20-->Thr was found to be principally responsible for this alteration. The second mutation cluster was responsible for increased binding of v-Rel to all the kappaB sites examined presumably because it stabilized v-Rel homodimers. This alteration in DNA binding was mapped to the group of two mutations within the cluster. In contrast, the third mutation cluster in the C-terminus of v-Rel destabilized the binding of v Rel to all of the kappaB sites examined. This is the first indication that regions outside the Rel Homology Region can participate in the control of binding of the c-Rel protein to DNA. The three mutation clusters examined contributed to the tumorigenic potential of v-Rel with the relative strength decreasing with their position from the N-terminus to the C-terminus. These results suggest that the oncogenic mutations in v-Rel cooperate and enable v-Rel to form nuclear complexes with aberrant DNA-binding properties that may directly alter gene expression and DNA replication resulting in the transformation of the cell. PMID- 9205097 TI - Epstein-Barr virus-encoded LMP1 and CD40 mediate IL-6 production in epithelial cells via an NF-kappaB pathway involving TNF receptor-associated factors. AB - Expression of the Epstein-Barr virus (EBV) transforming LMP1 in B cells activates the transcription factor NF-kappaB and induces phenotypic changes through two distinct domains in the cytoplasmic C-terminus of the protein. The aa 187-231 domain of LMP1, which is important for growth transformation, binds tumour necrosis factor (TNF) receptor associated factor (TRAF) 1 and TRAF3 and this interaction mediates subsequent signalling events. The TRAFs also associate with CD40, a member of the TNFR family, which upon ligation activates NF-kappaB and induces phenotypic changes similar to those mediated by LMP1. This study demonstrates that LMP1 expression in carcinoma cell lines and SV40-transformed keratinocytes results in induction of the pleiotropic cytokine interleukin 6 (IL6), an effect which is also observed upon CD40 ligation. The mechanism by which either LMP1 expression or CD40 ligation induces IL6 production was found to be NF-kappaB-dependent. Mutational analysis identified domains in the C-terminus of LMP1 which are important for NF-kappaB activation and IL6 secretion. LMP1 and CD40 share a common PxQxT core TRAF binding motif and mutations in or adjacent to this sequence impaired the ability of LMP1 or CD40 to induce NF-kappaB activation and IL6 secretion. The importance of TRAF interactions in mediating these effects was confirmed using dominant negative TRAF2 and TRAF3 mutants which also identified differences in the signalling events mediated by the two NF-kappaB activating domains of LMP1. A20, an anti-apoptotic protein which interacts with TRAF2 and blocks CD40-mediated NF-kappaB activity, also blocked NF-kappaB and IL6 secretion in LMP1-transfected epithelial cells. These results suggest that LMP1 regulates IL6 production in epithelial cells in a manner similar to CD40 ligation and implicate TRAFs as common mediators in the transduction of signals generated via the CD40 and LMP1 pathways. As a role for IL6 in regulating epithelial cell growth has previously been suggested, the control of IL6 secretion via the CD40 and LMP1 pathways may have implications for the growth of both normal and transformed epithelial cells. PMID- 9205098 TI - Chimeric oncoprotein E2a-Pbx1 induces apoptosis of hematopoietic cells by a p53 independent mechanism that is suppressed by Bcl-2. AB - The chimeric oncoprotein E2a-Pbx1 results from fusion of the E2A and PBX1 genes following t(1;19) chromosomal translocations in B cell precursor acute leukemias. Experimentally B cell progenitors do not tolerate constitutive expression of E2a Pbx1 which contrasts with transformation of several other cell types following its stable expression both in vitro and in vivo. To further investigate the effects of E2a-Pbx1 on the B cell progenitors, we conditionally expressed E2a Pbx1 under control of a metal response element in hematopoietic precursor cell lines in vitro. Inducible expression of E2a-Pbx1 resulted in cell death with the morphologic and molecular features of apoptosis. A structure-function analysis demonstrated that induction of apoptosis was not a dominant-negative effect of the E2a moiety but, rather, required the DNA-binding homeodomain of Pbx1. E2a Pbx1-induced apoptosis proceeded through a BCL2-responsive checkpoint eventuating in PARP inactivation but did require p53. Constitutive expression of E2a-Pbx1 did not induce apoptosis or continued cycling of Rat-1 fibroblasts in low serum conditions. These studies demonstrate that E2a-Pbx1 initiates programmed cell death of hematopoietic precursers by a mechanism that requires its chimeric transcriptional properties, but, unlike other nuclear oncoproteins, is independent of p53. PMID- 9205099 TI - Comprehensive allelotyping of human hepatocellular carcinoma. AB - Hepatocellular carcinoma (HCC) is one of the most common cancers in many parts of the world, however the molecular mechanisms underlying liver cell transformation remain obscure. A genome-wide scan of loss of heterozygosity (LOH) in tumors provides a powerful tool to search for genes involved in neoplastic processes. To identify recurrent genetic alterations in liver tumors, we examined DNAs isolated from 120 HCCs and their adjacent non tumorous parts for LOH using a collection of 195 microsatellite markers located roughly every 20 cM throughout 39 autosomal arms. The mean heterozygosity was 73%. Our findings provide additional support that LOH for loci on chromosomal arms 1p, 4q, 6q, 8p, 13q and 16p is significantly elevated in HCC. The highest percentage of LOH is found for a locus in 8p23 (42% of informative csaes). This corresponds to one of the most common genetic abnormalities reported to date in these tumors. In addition, high ratio of LOH (> or = 35%) is observed on chromosome arms which had not been implicated in previous studies, notably on 1q, 2q and 9q. No correlation was found between LOH of specific chromosomal regions and etiologic factors such as chronic infections with hepatitis B or C viruses. This first report of an extensive allelotypic analysis of HCC should help in identifying new genes whose loss of function contributes to the development of liver cancer. PMID- 9205100 TI - HMGIC, the gene for an architectural transcription factor, is amplified and rearranged in a subset of human sarcomas. AB - Amplified segments of the long arm of chromosome 12 are frequently observed in human sarcomas. In most cases there are separate amplified regions around the MDM2 and CDK4 genes. Here we show recurrent amplification of a third region encompassing HMGIC, a human architectural transcription factor gene. Reduced amplification frequency of sequences flanking the gene was observed, indicating that inclusion of this third region in the amplicons is also selected for. In three samples only the 5' part of HMGIC was amplified, suggesting preferential loss of the 3' part of the gene preceding or during amplification. In several other samples rearrangement of the gene was observed. Expression analysis showed transcripts of aberrant sizes, lacking 3' sequences, and 3' RACE of one sample revealed replacement of exons 4 and 5 with ectopic sequences. This truncation of HMGIC resembles that reported for translocations of HMGIC in benign tumors, including lipomas, and it is striking that the gene was frequently amplified or rearranged in well differentiated liposarcomas, the malignant counterpart of lipomas. It seems conceivable that high levels of either full length or truncated hmgic could be relevant for the etiology of these tumors. PMID- 9205101 TI - ayk1, a novel mammalian gene related to Drosophila aurora centrosome separation kinase, is specifically expressed during meiosis. AB - A novel murine gene, designated ayk1, which encodes a putative serine/threonine kinase has been cloned and characterized. The predicted catalytic domain of the protein is highly similar to that of Drosophila aurora (62.9% identity), and to that of Saccharomyces Ipl1 (49.4% identity). All three proteins also have very basic calculated isoelectric points (higher than 10). aurora has been recently shown to be crucial for centrosome separation and chromosome segregation, while Ipl1 is essential for yeast viability and accurate chromosome segregation. The results of Northern analysis and in situ RNA localization support a similar role for ayk1. The gene is specifically expressed in meiotically active cells, and during spermatogenesis, ayk1 transcripts accumulate just before the first meiotic division. Much lower levels are found in mitotically active cells. We propose that Ayk1, aurora and Ipl1 belong to a distinct new subfamily of kinases. These results suggest that the pathways controlling chromosome segregation are evolutionary conserved, and that similar control mechanisms operate in mitosis and meiosis. PMID- 9205102 TI - Expression of intracisternal A-particle retrotransposons in primary tumors of oncogene-expressing transgenic mice. AB - Intracisternal A-Particle (IAP) sequences are endogenous retrovirus-like mobile elements, present at 1000 copies in the mouse genome. These elements transpose in a replicative manner via an RNA intermediate and its reverse transcription, and their transposition should therefore be tightly controlled by their transcription level. The in vivo pattern of expression of these potentially mutagenic elements had previously been analysed in normal mice, and we have now investigated their expression in transgenic mice carrying different oncogenes (e.g. c-myc, v-Ha-ras, SV40 T-antigen) under tissue-specific promoters and disclosing tumors within the brain, the mammary or salivary glands, or the lymphoid organs. Northern blot analysis of IAP expression within the resulting tumors demonstrates a lack of significant and/or systematic effect of v-Ha-ras and SV40 T-antigen expression, but a systematic IAP induction in the myc-induced tumors. In this case, however, analysis of double transgenic mice obtained by crossing the tumor-prone mice with previously described transgenic mice carrying IAP reporter genes did not provide any evidence for induction of the IAP transgenes, therefore strongly suggesting that c-myc expression had an effect on only a limited number of IAP sequences- most probably depending on their position and/or methylation state. These results strengthen the importance of in vivo studies for a correct appraisal of complex biological processes, and moderate previous conclusions derived from in vitro analyses on the general activation of IAPs by oncogenes and on the role of these transposable elements in tumorigenesis. PMID- 9205103 TI - The transcription factor E2F-1 modulates TGF-beta1 RNA expression in glial cells. AB - The cell type specificity of the regulation of expression of the potent growth inhibitory cytokine transforming growth factor-beta (TGF-beta), prompted our analyses of the regulation of TGF-beta1 gene expression in glial cells by viral and cellular oncoproteins. We have shown that SV40 T-antigen diminished TGF-beta1 expression in glial cells and this repression was dependent on the ability of T antigen to interact with the tumor suppressor protein, pRb, and two structurally related proteins, p107 and p130. The cellular transcription factor E2F-1, which is a downstream effector of T-antigen, was unable to influence expression from the TGF-beta1 promoter by itself. Interestingly, E2F-1 could overcome viral T antigen-mediated repression of the TGF-beta1 promoter, suggesting potential feedback loop between TGF-beta and E2F in virally transformed glial cells. Using deletion analyses, we have mapped two E2F-1-responsive regions on the TGF-beta1 promoter: a T-antigen-dependent negative regulatory sequence (TdNRS) between -323 and -175, and a T-antigen-independent positive regulatory sequence (TiPRS) between -34 and +10 on the TGF-beta1 promoter. Further examination of TiPRS revealed the presence of a functional E2F binding site. Interestingly, the amino terminus of E2F-1 was required for its activation of TGF-beta1 expression, as mutations in that domain abolished the ability of E2F-1 to increase TGF-beta1 expression. These data suggest that yet-uncharacterized interaction between the amino terminus of E2F-1 and cellular proteins regulates TGF-beta1 expression. The mechanism for E2F-1-mediated T-antigen-dependent regulation of TGF-beta1 expression from TdNRS awaits further characterization. PMID- 9205104 TI - Anti-cell death activity promotes pulmonary metastasis of melanoma cells. AB - Bcl-2 inhibits apoptosis from a variety of stimuli, and a Bcl-2-binding protein BAG-1 also functions in protection from apoptosis in concert with Bcl-2. Here, we provide evidence that prolonged cell survival introduced by overexpression of Bcl 2 or BAG-1 proteins strongly promotes experimental pulmonary metastasis of melanoma B16-BL6 cells. In murine melanoma cell line B16-BL6, gene transfer mediated expression of the Bcl-2 or BAG-1 led to prolonged cell survival against serum-starved apoptosis in vitro. The Bcl-2-expressing B16 cells, B16-Bcl-2 and the BAG-1-expressing B16 cells, B16-BAG-1 strongly enhanced pulmonary metastasis in allogenic BALB/c nude mice and whole lung weights were increased by 2.4-fold and 1.4-fold, respectively, compared with control transfectants, suggesting that Bcl-2 is a stronger positive modulator of metastasis. When the viable B16-Bcl-2 and control transfectants were injected subcutaneously into BALB/c nude mice, the colony numbers of pulmonary metastasis of the B16-Bcl-2 transfectant increased by 5.6-fold compared with the control transfectants. These enhanced metastatic potentials in the B16-Bcl-2 and the B16-BAG-1 transfectants were well correlated with anti-cell death activity against serum-starvation and enhanced cell viability on limiting dilution. Analysis of the transfectants however revealed that their growth rates, invasive ability and cell motility were not significantly altered by overexpression of either Bcl-2 or BAG-1 proteins. Taken together, these studies demonstrate that prolonged cell survival is a crucial factor to promote metastasis of melanoma, thereby contributing to tumor progression. PMID- 9205105 TI - Identification of a 1300 kilobase deletion unit on chromosome 7q31.3 in invasive epithelial ovarian carcinomas. AB - We have used polymerase chain reaction (PCR) amplification of tandem repeats to study the pattern of allelic loss on chromosome 7q31 in invasive epithelial ovarian carcinomas and borderline ovarian tumors. Using 13 primer sets spanning loci from 7q31 to 7q32, 16 borderline and 54 invasive ovarian tumor tissue, together with their corresponding normal tissue, were analysed. Invasive epithelial ovarian tumors demonstrated loss of heterozygosity (LOH) at one or more loci on 7q in 32 of 54 cases (59%). The invasive epithelial ovarian tumors demonstrated the highest percentage of LOH at the loci D7S643 (20 of 40 informative cases, 50%) and GATA44F09 (18 of 42 informative cases, 43%). In contrary, only one borderline ovarian tumor showed LOH at one locus (GATA44F09, one of 14 informative cases, 7%). Our results display a sharp contrast in the pattern of LOH between invasive and borderline ovarian tumors suggesting that allelic loss at chromosome 7q31.3 may be involved in the development and progression of invasive epithelial ovarian tumors but not borderline ovarian tumors. Further analysis of the deletion map for the invasive epithelial ovarian tumors showed two regions likely to harbor ovarian tumor suppressor genes including a novel 1300 kilobase common loss region flanked by GATA44F09 and D7S643. PMID- 9205106 TI - Preferential activation of Fgf8 by proviral insertion in mammary tumors of Wnt1 transgenic mice. AB - Mouse mammary tumor virus (MMTV) is an insertional mutagen that has been demonstrated to transcriptionally activate flanking cellular proto-oncogenes. Previously we have used MMTV infection to accelerate mammary tumorigenesis in Wnt1 transgenic mice in order to identify genes that cooperate with the Wnt1 oncogene. Initial investigations into the resulting tumor collection, screened primarily by Southern analysis, showed that three fibroblast growth factor genes, Fgf8, Fgf3 and Fgf4, sustain activating insertion mutations in 10%, 42% and 6% of the tumors, respectively. Here, in an examination of the tumors from MMTV infected Wnt1 transgenic mice that emphasizes Northern analysis, we report transcriptional activation of Fgf8 in 30 additional tumors (increasing the percentage of activations to 50%), while no significant changes in the activation frequency of Fgf3 or Fgf4 were found. To determine the frequency of insertional activation in normal mice, we examined tumors from MMTV-infected nontransgenic littermates of the Wnt1 transgenics and from MMTV-infected BALB/c mice. Fgf8, Fgf3 and Fgf4 were found to be activated in 11%, 80% and 5%, respectively, of the tumors in the combined nontransgenic groups. Thus, there appears to be an increased predisposition for Fgf8 activations in Wnt1 transgenic mice versus normal mice, suggesting that cells expressing Wnt1 are especially sensitized to stimulation by FGF8 compared with FGF3 or FGF4. In contrast, the activation frequency of Fgf3 in tumors from MMTV-infected Wnt1 transgenic mice was approximately one-half that of normal mice. Our results show that this in vivo model of multistep tumorigenesis reveals significant differences in the activation rates of Fgf3 and Fgf8 depending upon the status of Wnt1 expression in the mammary gland. The differential activation of these Fgfs may relate to differences in their signaling pathways. PMID- 9205107 TI - Cloning and gene mapping of the chromosome 13q14 region deleted in chronic lymphocytic leukemia. AB - Frequent deletions and loss of heterozygosity in a segment of chromosome 13 (13q14) in cases of B-cell chronic lymphocytic leukemia (CLL) have suggested that this malignancy is caused by inactivation of an unknown tumor suppressor gene located in this region. Toward the identification of the putative CLL tumor suppressor, we have constructed a high-resolution physical map of YAC, PAC, and cosmid contigs covering 600 kb of the 13q14 genomic region. In addition to densely positioned genetic markers and STSs, this map was further annotated by localization of 32 transcribed sequences (ESTs) using a combination of exon trapping, direct cDNA selection, sample sequencing of cosmids and PACs, and homology searches. On the basis of these mapping data, allelic loss analyses at 13q14 using CLL tumor samples allowed narrowing of the genomic segment encompassing the putative CLL gene to <300 kb. Twenty-three ESTs located within this minimally deleted region are candidate exons for the CLL-associated tumor suppressor gene. PMID- 9205108 TI - Human gamma-aminobutyric acid-type A receptor alpha5 subunit gene (GABRA5): characterization and structural organization of the 5' flanking region. AB - The gamma-aminobutyric acid-type A receptor alpha5 subunit gene (GABRA5) is widely expressed in brain and localized to the imprinted human chromosome 15q11 q13. A combination of cDNA library screening and 5' RACE analysis led to identification of three distinct mRNA isoforms of GABRA5 in human adult and fetal brain tissues, each of which differs only in the noncoding 5' UTR sequence. Alignment of the genomic and cDNA sequences of GABRA5 revealed that the mRNA isoforms resulted from three alternative first exons 1A, 1B, and 1C. Northern blot analysis showed that the expression of GABRA5 was not only tissue specific but region specific in brain. CAT reporter assays revealed promoter elements in the 5' proximity of each first exon. The GABRA5 promoter regions lacked TATA and CCAAT boxes but contained several other consensus transcriptional factor recognition sequences. These findings suggest that the differential exon 1 usage of GABRA5 arises as a consequence of alternative promoter activation. PMID- 9205109 TI - The structure and organization of the human NPAT gene. AB - Ataxia telangiectasia (AT) is an autosomal recessive gene disorder, and ATM, a housekeeping gene, has been identified as the gene responsible for AT. Recently we found that another housekeeping gene, NPAT, is located upstream of ATM on human chromosome 11. The two housekeeping genes are transcribed in opposite directions and share a 0.5-kb 5' flanking sequence. The structure and organization of NPAT were determined by direct sequencing of cosmid clones carrying the gene and by application of the long and accurate (LA)-PCR method to amplify regions encompassing the exon/intron boundaries and all of the exons. The gene spans at least 44 kb and consists of 18 exons and 17 introns. It has been suggested that AT heterozygotes have an increased risk of developing cancer, especially breast cancer in women. Frequently, loss of heterozygosity at loci on 11q22-q24 has been observed in DNA isolated from tumors of the breast, uterine cervix, and colon, perhaps suggesting the location of a tumor suppressor gene in 11q22-q24. For investigation of the role of NPAT in AT and these tumors with allelic loss of 11q22-q24, appropriate primer sequences and PCR conditions for amplification of all the NPAT exons from genomic DNA were determined. We previously reported that no recombinations are found among Atm, Npat, and Acat1 (acetoacetyl-CoA thiolase) loci as determined by fine genetic linkage mapping of the mouse AT region. The results of the LA-PCR analysis using NPAT- and ACAT specific primers and human genomic DNA allowed us to map ACAT 12 kb centromeric to NPAT. PMID- 9205110 TI - A new dominant retinal degeneration (Rd4) associated with a chromosomal inversion in the mouse. AB - An autosomal dominant retinal degeneration, called Rd4, was found in a stock carrying the inversion In(4)56Rk, which was induced in a DBA/2J male. The inversion encompasses nearly all of Chromosome 4. It is homozygous lethal and in heterozygotes is always associated with retinal degeneration. In affected mice, the retinal outer nuclear and plexiform layers begin to reduce at 10 days of age, showing total loss at 6 weeks. The recordable electroretinograms (ERG) showed poorly at 3 to 6 weeks and were barely detected after 6 weeks of age. Retinal vessel attenuation, pigment spots, and optic atrophy appeared in the fundus at 4 weeks of age. Rd4 has not recombined with the inversion in an outcross, suggesting that the Rd4 locus is located very close to or is disrupted by one of the breakpoints of the inversion, either near the centromere or near the telomere. A human homolog would be expected to be located on human chromosomes 1p or 8q. PMID- 9205111 TI - Structures of the human gene for the protein disulfide isomerase-related polypeptide ERp60 and a processed gene and assignment of these genes to 15q15 and 1q21. AB - ERp60 (also known as ERp61 or GRP58) is an isoform of protein disulfide isomerase (PDI) that has two thioredoxin-like domains a and a' in positions corresponding to those of domains a and a' in the PDI polypeptide and shows a significant amino acid sequence similarity to PDI in almost all parts. We report here that the human ERp60 gene is about 18 kb in size and consists of 13 exons. No distinct correlation was found between its exon-intron organization and the modular structure of the ERp60 polypeptide, nor were any similarities in exon-intron organization found between the human ERp60, PDI, and thioredoxin genes. The 5' flanking region of the ERp60 gene has no TATAA box or CCAAT motif but contains several potential binding sites for transcription factors. The highest levels of expression of the ERp60 mRNA were found by Northern blotting in the liver, placenta, lung, pancreas, and kidney, and the lowest in the heart, skeletal muscle, and brain. We also isolated an intronless ERp60 gene that probably represents a pseudogene. The ERp60 gene was mapped by fluorescence in situ hybridization to 15q15 and the processed gene to 1q21, so that neither was located on the same chromosome as the human PDI and thioredoxin genes. PMID- 9205113 TI - Inactivation of the mouse HPRT locus by a 203-bp retroposon insertion and a 55-kb gene-targeted deletion: establishment of new HPRT-deficient mouse embryonic stem cell lines. AB - To obtain useful hypoxanthine phosphoribosyl-transferase (HPRT)-deficient mouse ES cell lines, two different methods were employed: (i) selection of spontaneous 6-TG-resistant mutants and (ii) gene targeting of the HPRT locus. The first approach resulted in the establishment of E14.1TG3B1, a spontaneous HPRT deficient cell line with an insertional mutation of 203 bp in the third exon of the HPRT gene. The insert is highly homologous to the B2 mouse repetitive element and has all the expected retroposon characteristics, thus providing an example of gene inactivation by retroposon insertion. This clone exhibited stable 6-TG resistance and high germ-line transmission frequency. Thus E14.1TG3B1 is a useful ES cell line for modifying the mouse genome using the HPRT gene as a selection marker and for transmission at a high frequency into the mouse germ line. The second approach resulted in a 55-kb deletion of the mouse HPRT locus, demonstrating the feasibility of replacement-targeting vectors to generate large genomic DNA deletions. PMID- 9205112 TI - A transcript map encompassing the multiple endocrine neoplasia type-1 (MEN1) locus on chromosome 11q13. AB - A transcription map of a 1200-kb region encompassing the MEN1 locus was constructed by direct cDNA selection and mapping ESTs. A total of 29 genes were mapped. Ten transcripts were identified by cDNA selection of a focused 300-kb genomic region telomeric to the MEN1 consensus region. Since many of the sequences cloned by cDNA selection also identified ESTs from the region, 19 additional RH-mapped ESTs were mapped to the entire contig region by PCR amplification of genomic clones. Nine known genes, 2 putative human homologues to mouse genes, and 18 novel transcripts map to the region. Transcripts that map to the MEN1 interval PYGM-D11S449 include SGC35223, IB1256, AA147620, ZFM1, FAU, and CAPN1. The latter 3 known genes have already been excluded as candidate MEN1 genes. The 2 putative human homologues of mouse genes Ltbp2 and Spa-1 may be candidate tumor suppressor genes, but they map telomeric to D11S449. Although both of these genes map outside the MEN1 consensus region they may play a role in sporadic endocrine tumors independent of the MEN1 gene or in other tumors, such as breast cancer, that have loss of heterozygosity within this region. PMID- 9205114 TI - Physical mapping 220 kb centromeric of the human MHC and DNA sequence analysis of the 43-kb segment including the RING1, HKE6, and HKE4 genes. AB - A cosmid contig was constructed from a YAC clone with a 220-kb insert that spans the centromeric side of the human MHC class II region, corresponding to the mouse t complex. The gene order was identified to be HSET-HKE1.5-HKE2-HKE3-RING1-HKE6- HKE4 (RING5). The genomic sequence of a 42,801-bp long region encoded by one cosmid clone in the RING1, HKE6, and HKE4 subregions was determined by the shotgun method. The exon-intron organization of these three genes, RING1 (Ring finger protein), HKE6 (steroid dehydrogenase-like protein), and HKE4 (transmembrane protein with histidine-rich charge clusters), was determined. The previously reported RING2 gene was revealed to be identical to HKE6. Transcripts from HKE4 were detected in the placenta, lung, kidney, and pancreas. Those of HKE6 were found in the liver and pancreas. The 25-kb region proximal to the RING1 gene includes an extensive dense cluster of Alu repeats (about 1.2 Alu per kb), and no gene has been identified in this so far. The region is equivalent to part of the mouse t complex and could be of relevance to human development. PMID- 9205115 TI - A 2.8-Mb clone contig of the multiple endocrine neoplasia type 1 (MEN1) region at 11q13. AB - Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant disorder that results in parathyroid, anterior pituitary, and pancreatic and duodenal endocrine tumors in affected individuals. The MEN1 locus is tightly linked to the marker PYGM on chromosome 11q13, and linkage analysis has placed the MEN1 gene within a 2-Mb interval flanked by D11S1883 and D11S449. As a step toward cloning the MEN1 gene, we have constructed a 2.8-Mb clone contig consisting of YAC and bacterial clones (PAC, BAC, and P1) for the D11S480 to D11S913 region. The bacterial clones alone represent nearly all of the 2.8-Mb contig. The contig was assembled based on a high-density STS-content analysis of 79 genomic clones (YAC, PAC, BAC, and P1) with 118 STSs. The STSs included 22 polymorphic markers and 20 transcripts, with the remainder primarily derived from the end sequences of the genomic clones. An independent cosmid contig for the 1-Mb PYGM-SEA region was also generated. Support for correctness of the 2.8-Mb contig map comes from an independent ordering of the clones by fiber-FISH. This sequence-ready contig will be a useful resource for positional cloning of MEN1 and other disease genes whose loci fall within this region. PMID- 9205116 TI - Cloning and characterization of a novel member of the human Mad gene family (MADH6). AB - MAD (mothers against decapentaplegic)-related proteins (MADRs) are intracellular components that play critical roles in signal-transduction pathways involving the transforming growth factor beta (TGFbeta) superfamily. Some Mad genes are candidates for tumor-suppressor functions. From a human fetal brain cDNA library we have isolated a novel Mad-related gene. Two alternatively transcribed mRNAs encode deduced 430- and 467-amino-acid peptides that showed high levels of similarity to MADR1/Smad1/hMAD1 (about 80% identity at the amino acid level). This gene, which we designated MADH6, resides on 13q12-q14 between BRCA2 and RB, a region that frequently displays loss of heterozygosity in breast, liver, and prostate cancers. PMID- 9205117 TI - The genomic organization of the murine Pax 8 gene and characterization of its basal promoter. AB - Lambda phage clones containing the murine Pax 8 gene were isolated from a C57BL/6 kidney genomic mouse library using mouse cDNA fragments as probes. A clone encompassing about 16 kb of the 5' untranslated region of the murine Pax 8 gene was isolated from a mouse embryonic stem cell (D3) library. The murine Pax 8 gene has a size of approximately 26 kb and contains the coding sequence for mRNA in 12 exons. The major and several minor transcription initiation sites were identified. Position +1 is located 488 nucleotides upstream of the ATG initiation codon and 24 bases downstream of a TATA-like sequence, ATAAAA. The translation initiation and termination sites are located in exons 2 and 12, respectively. Further analysis of 570 bases of the 5' flanking sequence revealed AP2, SP1, PEA3, zeste, NF-kappaB, and CCAAT consensus binding sites. Ribonuclease protection assays with a probe spanning the first two exons of mouse Pax 8 cDNA on total RNA samples isolated from different tissues of newborn mice show that the murine Pax 8 gene is predominantly expressed in kidney tissue. Low levels of Pax 8 gene expression were also found in the liver, spleen, lung, brain, and heart. The same transcription initiation sites are utilized in different tissues of newborn mice and embryo at Day 10.5 postconception. A FISH assay shows that the murine Pax 8 gene is located on chromosome 2, map position B. PMID- 9205118 TI - Cloning and chromosomal localization of a gene (GPR18) encoding a novel seven transmembrane receptor highly expressed in spleen and testis. AB - Using the techniques of relaxed stringency polymerase chain reaction and genomic library screening, we have isolated homologous canine and human genes that encode a novel putative seven transmembrane G-protein-linked receptor. The gene encodes an open reading frame (ORF) of 993 bp. The sequences of the canine and human ORFs are highly conserved, sharing 89% nucleotide identity and 92% amino acid similarity between the two species. Northern blot analysis demonstrates that mRNA transcripts of the gene are abundantly expressed in testis and spleen with a lesser degree of expression observed in several other tissues associated with endocrine and immunologic/hematologic function. The gene, designated GPR18, was localized to human chromosome 13q32 using fluorescence in situ hybridization. PMID- 9205119 TI - Construction and characterization of a Plasmodium vivax genomic library in yeast artificial chromosomes. AB - Here we describe the construction of a representative YAC library for the human malarial parasite Plasmodium vivax. As P. vivax cannot be maintained continuously under laboratory conditions, the P. vivax DNA necessary for the library construction was isolated from a single human patient presenting himself with vivax malaria to a local hospital in the Brazilian Amazon. Thus, this YAC library is the first of its kind to be generated from patient-derived material. The YAC library consists of 560 clones with an average insert size of 180 kb. Of 9 published P. vivax genes, 8 were found to be present in the library. In addition, 12 P. vivax telomeric YAC clones were identified. PMID- 9205120 TI - LTW4 protein on mouse chromosome 1 is a member of a family of antioxidant proteins. AB - Based on its map position, polymorphism pattern, and expression in the kidney, the gene encoding liver 20,000-30,000 MW protein 4 (LTW4) can be considered a potential candidate for the Jckm2 modifying locus, which mediates the severity of polycystic kidney disease in the juvenile cystic kidney mouse. Using two dimensional gel electrophoresis, we identified variants of a 26-kDa polypeptide that differed in their isoelectric points between the C57BL/6J and the DBA/2J inbred strains in a pattern similar to that originally described for LTW4 protein. N-terminal amino acid sequence was obtained by microsequencing analysis, and full-length clones were obtained by RT-PCR amplification and characterized. The map position of the cloned gene was determined and corresponded to that previously described for Ltw4. The gene has homology to a class of proteins characterized as thiol-specific antioxidants that are protective against damage caused by oxidative stress. The murine MER5 gene is also a member of this gene family and has recently been renamed Antioxidant protein 1 (Aop1), based on its functional characterization. We therefore propose that the gene encoding LTW4 be called Aop2. PMID- 9205121 TI - Reln(rl-Alb2), an allele of reeler isolated from a chlorambucil screen, is due to an IAP insertion with exon skipping. AB - The reeler Albany2 mutation (Reln(rl-Alb2) in the mouse is an allele of reeler isolated during a chlorambucil mutagenesis screen. Homozygous animals had drastically reduced concentrations of reelin mRNA, in which an 85-nt exon was absent. At the genomic level, the mutation was shown to be due to an intracisternal A-particle insertion leading to exon skipping. This appears to be the first observation of retrotransposon insertion during chlorambucil mutagenesis. PMID- 9205122 TI - Molecular cloning of a novel vascular endothelial growth factor, VEGF-D. AB - We have identified and characterized a novel vascular endothelial growth factor (VEGF), VEGF-D, which is structurally related to vascular endothelial growth factor C. A full-length cDNA for human VEGF-D was cloned following the identification of an EST obtained through a TFASTA search of public EST databases. The murine VEGF-D was subsequently isolated from a mouse lung cDNA library. The human VEGF-D gene was mapped to human chromosome Xp22.31. Both human and mouse VEGF-D are strongly expressed in lung and encode the eight cysteine residues that are highly conserved among the members of this family. The high level of conservation between mouse and human VEGF-D may emphasize the biological importance of this gene. Recently the murine gene, FIGF, which is identical to mouse VEGF-D, was reported. PMID- 9205123 TI - Construction of an integrated physical and gene map of human chromosome 20p12 providing candidate genes for Alagille syndrome. AB - Physical mapping and localization of eSTS markers were used to generate an integrated physical and gene map covering a approximately 10-Mb region of human chromosome 20p12 containing the Alagille syndrome (AGS) locus. Seventy-four STSs, 28 of which were derived from cDNA sequences, mapped with an average resolution of 135 kb. The 28 eSTS markers define 20 genes. Six known genes, namely CHGB, BMP2, PLCB1, PLCB4, SNAP, and HJ1, were precisely mapped. Among the genes identified, one maps in the smallest region of overlap of the deletions associated with AGS and could therefore be regarded as a candidate gene for Alagille syndrome. PMID- 9205124 TI - Cloning, mRNA expression, and chromosomal mapping of mouse and human preprocortistatin. AB - Cortistatin is a 14-residue putative neuropeptide with strong structural similarity to somatostatin and is expressed predominantly in cortical GABAergic interneurons of rats. Administration of cortistatin into the brain ventricles specifically enhances slow-wave sleep, presumably by antagonizing the effects of acetylcholine on cortical excitability. Here we report the identification of cDNAs corresponding to mouse and human preprocortistatin and the mRNA distribution and gene mapping of mouse cortistatin. Analysis of the nucleotide and predicted amino acid sequences from rat and mouse reveals that the 14 C terminal residues of preprocortistatin, which make up the sequence that is most similar to somatostatin, are conserved between species. Lack of conservation of other dibasic amino acid residues whose cleavage by prohormone convertases would give rise to additional peptides suggests that cortistatin-14 is the only active peptide derived from the precursor. As in the rat, mouse preprocortistatin mRNA is present in GABAergic interneurons in the cerebral cortex and hippocampus. The preprocortistatin gene maps to mouse chromosome 4, in a region showing conserved synteny with human 1p36. The human putative cortistatin peptide has an arginine for lysine substitution, compared to the rat and mouse products, and is N terminally extended by 3 amino acids. PMID- 9205125 TI - Identification, localization, and expression of two novel human genes similar to deoxyribonuclease I. AB - Two novel cDNAs, DNAS1L2 and DNAS1L3, are predicted to encode proteins of 299 and 305 amino acids with 56 and 46% residue identity (71 and 63% similarity), respectively, to deoxyribonuclease I (DNase I). DNAS1L2 is located on a 16p13.3 cosmid, while DNAS1L3 maps to 3p14.3-p21.1 by fluorescence in situ hybridization and by PCR analysis of a radiation hybrid panel. Northern analysis revealed DNAS1L3 expression nearly exclusively in liver, while DNAS1L2 expression was detected in brain by RT-PCR. The previously defined DNL1L or DNAS1L1 is expressed highest in heart and skeletal muscle, while DNase I is expressed in the pancreas, parotid gland, and kidney. Thus, to date, four DNase I-like genes that show different tissue expression patterns are known. A comparison of DNAS1L1, DNAS1L2, and DNAS1L3 with the well-characterized DNase I suggests that the DNAS1L proteins are unlikely to be glycosylated or bind actin; however, catalytic and calcium- and DNA-binding residues are conserved, and potentially cleavable signal peptides are present among all these proteins. This analysis also identifies regions of high conservation among these proteins with no currently assigned function. PMID- 9205126 TI - Genomic organization and primary characterization of miap-3: the murine homologue of human X-linked IAP. AB - IAPs (inhibitor of apoptosis proteins) are a recently identified family of proteins that function in the cell death pathway to inhibit programmed cell death. We have earlier reported cloning of four human IAPs: NAIP, hiap-1, hiap-2, and xiap. To facilitate studies of these genes, we have undertaken the cloning of their murine homologues. We report here the cloning, mapping, and preliminary characterization (including cellular and tissue distribution profile) of the murine homologue of the human X-linked IAP (xiap). The mouse gene called miap-3 (for murine IAP-3; GenBank Accession No. nuc 1 U88990) has a coding region of 1.5 kb that encodes a protein of 55 kDa and has 87 and 94% homology with its human homologue at DNA and amino acid levels, respectively. Northern blot analysis reveals an 8-kb miap-3 transcript in all tissues examined to date. miap-3 is composed of six exons and five introns spanning approximately 20 kb. miap-3 has been assigned to the A3-A5 region of mouse chromosome X by FISH analysis. PMID- 9205127 TI - Isolation and chromosomal localization of GPR31, a human gene encoding a putative G protein-coupled receptor. AB - The screening of a human genomic library with a chemokine receptor-like probe allowed us to obtain a putative member of the G protein-coupled receptor gene (GPCR) family, designated GPR31. Its deduced amino acid sequence encodes a polypeptide of 319 amino acids that shares 25-33% homology with members of the chemokine, purino, and somatostatin receptor gene families. Amino acid sequence comparison reveals that the best match in the protein databases is with the human orphan GPCR called HM74 (33% identity). Southern genomic analysis of the GPR31 gene shows a hybridization pattern consistent with that of a single-copy gene. Using fluorescence in situ hybridization, we have determined the chromosomal and regional localization of the GPR31 gene at 6q27. The GPR31 mRNA is expressed at low levels by several human cell lines of different cellular origins. The phylogenetic analysis suggests that the GPR31 receptor may represent a member of a new GPCR subfamily. PMID- 9205129 TI - Genomic organization and complete nucleotide sequence of the human PWP2 gene on chromosome 21. AB - The human PWP2 gene is the human homologue of the yeast periodic tryptophan protein 2 (PWP2) gene and is a member of the gene family that contains tryptophan aspartate (WD) repeats. Genomic sequencing revealed that the human PWP2 gene consists of 21 exons spanning approximately 24 kb and locates just between the two genes EHOC-1 and KNP-I and distal to a NotI site of LJ104 (D21S1460) on chromosome 21q22.3. Analysis of the 5'-flanking DNA sequence revealed that the upstream region of the PWP2 gene is associated with a CpG island containing the NotI site of LJ104. Since PWP2 is considered to be a candidate for genetic disorders mapped in the 21q22.3 region, the information including nucleotide sequence and genomic organization of the PWP2 gene should be invaluable for the mutation analysis of the corresponding genetic disorders. PMID- 9205130 TI - The gene encoding protein 4.2 is distinct from the mouse platelet storage pool deficiency mutation pallid. AB - Previous studies identified the gene encoding the erythrocyte membrane protein 4.2 (Epb4.2) as a candidate for the mouse mutation pallid (pa); Epb4.2 genetically colocalized near pa on mouse Chromosome 2, and a truncated Epb4.2 transcript was present in tissues derived from pallid mice. We report here evidence that Epb4.2 and pa are not allelic. The pallid cDNA and intron/exon boundaries show no significant variation from the known BALB/c and C57BL/6J Epb4.2 sequence, and normal immunoreactive 72-kDa protein 4.2 is present in pallid tissues. Two recombinations between Epb4.2 and pa were identified in 173 phenotypically mutant (C57BL/6J-pa/pa x Mus castaneus) F2 animals. Northern blotting reveals a truncated Epb4.2 transcript in kidney mRNA from normal wild Mus domesticus (WSB/Ei) mice that comigrates with the pallid Epb4.2 mRNA. As the pa mutation originally arose in a wild M. domesticus mouse, we conclude that the Epb4.2 mRNA characteristic of pallid is a normal polymorphism derived from its wild ancestor and that Epb4.2 and pa are distinct loci. PMID- 9205131 TI - Chromosomal mapping of the PFTAIRE gene, Pftk1, a cdc2-related kinase expressed predominantly in the mouse nervous system. PMID- 9205128 TI - Chromosomal localization of three human dual specificity phosphatase genes (DUSP4, DUSP6, and DUSP7). AB - Mitogen-activated protein (MAP) kinase phosphatases constitute a growing family of dual specificity phosphatases thought to play a role in the dephosphorylation and inactivation of MAP kinases and are therefore likely to be important in the regulation of diverse cellular processes such as proliferation, differentiation, and apoptosis. For this reason it has been suggested that MAP kinase phosphatases may be tumor suppressors. We have determined the chromosomal locations of three human dual specificity phosphatase genes by fluorescence in situ hybridization and radiation hybrid mapping. The genes were localized to three different chromosomes, MKP2 (DUSP4) to 8p11-p12, MKP3 (DUSP6) to 12q22-q23, and MKPX (DUSP7) to 3p21. This will allow the potential roles of these genes in disease processes to be evaluated. PMID- 9205132 TI - Chromosomal localization of genes encoding CAN/Nup214-interacting proteins--human CRM1 localizes to 2p16, whereas Nup88 localizes to 17p13 and is physically linked to SF2p32. PMID- 9205133 TI - Assignment of HMAT1, the human homolog of the murine mammary transforming gene (MAT1) associated with tumorigenesis, to 1q21.1, a region frequently gained in human breast cancers. PMID- 9205134 TI - Msx2 is a transcriptional regulator in the BMP4-mediated programmed cell death pathway. AB - Homeobox-containing genes play an important role in patterning processes that occur during embryogenesis. Programmed cell death is a key process during pattern formation. The mechanisms by which programmed cell death is spatially regulated are not well characterized. Msx1 and Msx2 are two closely related homeobox containing genes that are expressed at sites where cellular proliferation and programmed cell death occur, including the developing limb and the cephalic neural crest. Tissue interactions are necessary for the maintenance of Msx1 and Msx2 expression and programmed cell death. It has been demonstrated that BMP4 can regulate cell death at these same sites as well as induce Msx expression. These observations lead to the hypothesis that Msx2 is a key regulator of cell death in the BMP-mediated pathway. Embryonic stem cell lines will undergo processes typical of early embryogenesis upon aggregation and have recently been shown to provide a model system for programmed cell death. In contrast to ES cells, we see that P19 cells do not undergo pronounced cell death upon aggregation; however, constitutive ectopic Msx2 expression in P19 cells results in a marked increase in apoptosis induced upon aggregation but has no effect when cells are grown as a monolayer. If aggregates are allowed to interact with a substrate, the process of programmed cell death is completely inhibited. Addition of BMP4 to aggregated P19 cells also results in cell death; however, BMP4 does not increase levels of cell death in Msx2-expressing cells. Addition of BMP4 to P19 cells results in an induction of Msx2 transcription consistent with its proposed role in cell death in the embryo. Our data support a model by which BMP4 induces programmed cell death via an Msx2-mediated pathway and provide direct functional evidence that Msx2 expression is a regulator of this process. PMID- 9205135 TI - Genetic analysis of stomatogastric nervous system development in Drosophila using enhancer trap lines. AB - The stomatogastric nervous system (SNS) of Drosophila melanogaster is a small, simply organized neural circuitry which innervates the anterior enteric system. It is responsible for regulating the passage of food through the pharynx and esophagus and into the midgut. Here we show that the development of the SNS is amenable to genetic dissection. We screened lines from a P-element mutagenesis, selecting those with lacZ reporter gene expression and/or a phenotype in the SNS, associated glia, and garland cells. We report a collection of expression patterns and mutant phenotypes among lines found to have a mutation in genes required for the establishment of the larval SNS. Our results indicate that SNS development depends on pattern organizer genes including components of the Ras/Raf pathway. PMID- 9205136 TI - Identification of metalloprotease/disintegrins in Xenopus laevis testis with a potential role in fertilization. AB - Proteins containing a membrane-anchored metalloprotease domain, a disintegrin domain, and a cysteine-rich region (MDC proteins) are thought to play an important role in mammalian fertilization, as well as in somatic cell-cell interactions. We have identified PCR sequence tags encoding the disintegrin domain of five distinct MDC proteins from Xenopus laevis testis cDNA. Four of these sequence tags (xMDC9, xMDC11.1, xMDC11.2, and xMDC13) showed strong similarity to known mammalian MDC proteins, whereas the fifth (xMDC16) apparently represents a novel family member. Northern blot analysis revealed that the mRNA for xMDC16 was only expressed in testis, and not in heart, muscle, liver, ovaries, or eggs, whereas the mRNAs corresponding to the four other PCR products were expressed in testis and in some or all somatic tissues tested. The xMDC16 protein sequence, as predicted from the full-length cDNA, contains a metalloprotease domain with the active-site sequence HEXXH, a disintegrin domain, a cysteine-rich region, an EGF repeat, a transmembrane domain, and a short cytoplasmic tail. To study a potential role for these xMDC proteins in fertilization, peptides corresponding to the predicted integrin-binding domain of each protein were tested for their ability to inhibit X. laevis fertilization. Cyclic and linear xMDC16 peptides inhibited fertilization in a concentration dependent manner, whereas xMDC16 peptides that were scrambled or had certain amino acid replacements in the predicted integrin-binding domain did not affect fertilization. Cyclic and linear xMDC9 peptides and linear xMDC13 peptides also inhibited fertilization similarly to xMDC16 peptides, whereas peptides corresponding to the predicted integrin-binding site of xMDC11.1 and xMDC11.2 did not. These results are discussed in the context of a model in which multiple MDC protein-receptor interactions are necessary for fertilization to occur. PMID- 9205137 TI - M-spondin, a novel ECM protein highly homologous to vertebrate F-spondin, is localized at the muscle attachment sites in the Drosophila embryo. AB - The muscle attachment site (MAS) in Drosophila provides a unique and excellent model system to study the mechanism of cell-matrix adhesion in developing organisms. Here, we report on the isolation and characterization of a novel extracellular matrix (ECM) molecule localized at the MAS, encoded by the M spondin (mspo) gene. M-spondin protein contains a thrombospondin type I repeat (TSR) previously found in a variety of ECM molecules. Furthermore, it shares two conserved domains with F-spondin, a vertebrate ECM molecule with TSRs. The presence of TSR(s) and the two homologous domains thus defines a novel gene family of ECM molecules. The mspo mRNA was expressed by a large subset of muscles in the embryonic body wall. Secreted M-spondin protein diffused and eventually became immobilized at the MAS in late embryos. When expressed in S2 cells, the protein was secreted and became concentrated in the matrix on the surface of the culture dish. Genetic analysis revealed that both deletion mutants and misexpression mutants suffered no obvious developmental defects. We propose that M-spondin, although its function is redundant, is a component of the ECM and mediates mechanical linkage between the muscles and apodemes. PMID- 9205138 TI - Development of mechanisms regulating intracellular Ca2+ concentration in cardiac muscle cells of early chick embryos. AB - The development of mechanisms for the regulation of intracellular-free calcium ion concentration ([Ca2+]i) was investigated in precardiac mesodermal cells (PMC) and cardiac muscle cells (CMC) from early chick embryos by microfluorometry using a Ca2+-sensitive fluorescent probe, fura-2, and transmission electron microscopy. Microfluorometry indicated that two types of regulatory mechanisms, involving the dihydropyridine receptor (DHPR) and the ryanodine receptor (RYR), are present in CMC when the heartbeat begins at the 8-9 somite stages. Nifedipine completely suppressed the beating of hearts isolated from embryos on Days 1.5 and 2. Ryanodine had no effect on the beating of hearts isolated from embryos on Day 1.5, though it completely suppressed beating in hearts from Embryonic Day 2. Microfluorometry revealed that a change occurred in the Ca2+-regulating mechanisms of CMC on Day 2. Transmission electron microscopy showed the appearance in CMC, also on Day 2, of peripheral couplings with feet structures, and SR adjacent to the Z-line of myofibrils. These findings suggest that the calcium-induced calcium-release (CICR) mechanism appears in the CMC of the chick on the second day of embryonic development. PMID- 9205139 TI - Intracellular injections of a soluble sperm factor trigger calcium oscillations and meiotic maturation in unfertilized oocytes of a marine worm. AB - How sperm trigger activating calcium transients in eggs remains a central, unresolved question in fertilization biology. To determine if a soluble sperm factor can generate a fertilization-like calcium response in the absence of sperm egg binding, aqueous extracts of sperm from the nemertean worm Cerebratulus lacteus were mixed with Ca2+-sensitive fluorescent dyes and injected into unfertilized, metaphase-I-arrested oocytes. Based on confocal imaging analyses, unfertilized oocytes that had been injected with sperm extract routinely produced oscillating Ca2+ waves and resumed meiotic maturation in a manner that closely resembled normal fertilization. Calcium oscillations and maturation were typically lacking in control oocytes that had been (i) injected with buffer alone or with buffer containing added calcium, (ii) given external treatments of the sperm factor, or (iii) injected with extracts made from cells other than sperm. Boiling or protease treatment essentially abolished the potency of the sperm extract, and nonboiled extracts retained full activity in >10-kDa fractions, but not in <10-kDa fractions. Collectively, such findings suggest that the sperm of C. lacteus possess a soluble protein that can bypass oolemmal surface receptors to act within the ooplasm as a trigger of repetitive Ca2+ waves and meiotic maturation. Results obtained in this study are discussed with respect to the minimum amount of extract needed for egg activation and whether the oscillogenic substance is sufficiently concentrated in a single sperm to play a biological role during fertilization. PMID- 9205140 TI - Neuroepithelial stem cells from the embryonic spinal cord: isolation, characterization, and clonal analysis. AB - Adherent cultures of E10.5 rat neuroepithelial cells (NEP cells) from the caudal neural tube require FGF (fibroblast growth factor) and CEE (chick embryo extract) to proliferate and maintain an undifferentiated phenotype in culture. Epidermal growth factor (EGF) does not support E10.5 NEP cells in adherent culture and NEP cells do not form EGF-dependent neurospheres. NEP cells, however, can be grown as FGF-dependent neurospheres. NEP cells express nestin and lack all lineage specific markers for neuronal and glial sublineages, retain their pleuripotent character over multiple passages, and can differentiate into neurons, astrocytes, and oligodendrocytes when plated on laminin in the absence of CEE. In clonal culture, NEP cells undergo self-renewal and generate colonies that vary in size from single cells to several thousand cells. With the exception of a few single cell clones, all other NEP-derived clones contain more than one identified phenotype, with over 40% of the colonies containing A2B5, beta-111 tubulin, and GFAP-immunoreactive cells. Thus, NEP cells are multipotent and capable of generating multiple neural derivatives. NEP cells also differentiate into motoneurons immunoreactive for choline acetyl transferase (ChAT) and the low affinity neurotrophin receptor (p75) in both mass and clonal culture. Double labeling of clones for ChAT and glial, neuronal, or oligodendrocytic lineage markers shows that motoneurons always arose in mixed cultures with other differentiated cells. Thus, NEP cells represent a common progenitor for motoneurons and other spinal cord cells. The relationship of NEP cells with other neural stem cells is discussed. PMID- 9205141 TI - Impaired lung branching morphogenesis in the absence of functional EGF receptor. AB - The mammalian lung develops through branching morphogenesis which is controlled by growth factors, hormones, and extracellular matrix proteins. We have evaluated the role of EGF-receptor signaling in lung morphogenesis by analyzing the developmental phenotype of lungs in mice with an inactivated the EGF-receptor gene both in vivo and in organ culture. Neonatal EGF-receptor-deficient mice often show evidence of lung immaturity which can result in visible respiratory distress. The lungs of these mutant mice had impaired branching and deficient alveolization and septation, resulting in a 50% reduction in alveolar volume and, thus, a markedly reduced surface for gas exchange. The EGF-receptor inactivation also resulted in type II pneumocyte immaturity, which was apparent from their increased glycogen content and a reduced number of lamellar bodies. The defective branching was already evident at Day 12 of embryonic development. When explants of embryonic lungs from Day 12 embryos were cultured under defined conditions, the branching defect in EGF-receptor-deficient lungs was even more pronounced, with only half as many terminal buds as normal lungs. EGF treatment stimulated the expression of surfactant protein C and thyroid transcription factor-1 in cultured normal lungs, but not in EGF-receptor-deficient lungs, suggesting that EGF-receptor signaling regulates the expression of these marker genes during type II pneumocyte maturation. Taken together, our data indicate that signal transduction through the EGF receptor plays a major role in lung development and that its inactivation leads to a respiratory distress-like syndrome. PMID- 9205142 TI - Gene transfer and thoracic surgery: more to come. PMID- 9205143 TI - Thoughts and considerations on modeling coronary bypass surgery risk. PMID- 9205144 TI - Cardiothoracic databases: where are we headed? PMID- 9205145 TI - Prospective study of the natural history of thoracic aortic aneurysms. AB - BACKGROUND: The decision whether or not to recommend resection of moderately large descending thoracic and thoracoabdominal aneurysms requires weighing the relatively high mortality and significant risk of paraplegia associated with operation against the likelihood that the aneurysm will rupture spontaneously, with an almost invariably fatal outcome. To better define the risk of aneurysm rupture, we undertook a prospective study of patients who had not had operation on their moderately large descending thoracic and thoracoabdominal aneurysms. METHODS: Patients were enrolled at the time of their second computed tomographic scans: three-dimensional computer-generated reconstructions allowed determination of several dimensional parameters for each study, including diameters and cross sectional areas at the site of maximal dilatation in the descending aorta and in the abdomen as well as total thoracoabdominal surface area. Comparisons of serial studies permitted calculation of yearly rates of change in these dimensions. RESULTS: Of 114 patients, 8 died of causes unrelated to the aneurysm, 26 died of rupture, 20 met previously determined criteria for operation, and 60 survived without operation or rupture. Multivariate regression analysis identified maximal diameter in the descending and in the abdominal aorta as independent risk factors for rupture, as well as older age, the presence of even uncharacteristic pain, and a history of chronic obstructive pulmonary disease. A piecewise exponential model enabled construction of an equation allowing calculation of rate of rupture in patients in whom the values of the risk factors are known, and also of the probability of rupture in a given individual over a specified time interval. CONCLUSIONS: Because using this equation--based on easily determined risk factors (age, pain, chronic obstructive pulmonary disease, maximal thoracic and maximal abdominal aortic diameter)--allows the risk of aneurysm rupture within a given interval to be estimated fairly accurately for each individual patient, it is our current practice to recommend operation when the calculated risk of rupture within 1 year exceeds the anticipated mortality of elective operation, rather than relying on general operative guidelines based almost exclusively on aneurysm size. PMID- 9205146 TI - Effects of nitric oxide after cardiac transplantation in the setting of recipient pulmonary hypertension. AB - BACKGROUND: Recipient pulmonary hypertension secondary to chronic congestive heart failure is a significant risk factor for right ventricular failure after cardiac transplantation. In this study, the hemodynamic and inotropic effects of nitric oxide (NO) were examined after bicaval cardiac transplantation in the setting of monocrotaline pyrrole-induced recipient chronic pulmonary hypertension. METHODS: Twenty dogs underwent 10 successfully completed transplantation experiments. Recipients underwent pulmonary artery injection of 3 mg/kg monocrotaline pyrrole 4 months before transplantation. Measurements were taken 1 hour after cessation of cardiopulmonary bypass and after NO inhalation. Pulmonary vascular impedance was calculated using Fourier analysis, and cardiac function was assessed with load-insensitive means (preload recruitable stroke work). RESULTS: At the time of transplantation, the precardiopulmonary bypass levels of pulmonary vascular resistance in recipient animals were significantly greater when compared with donor levels, and were further significantly increased after cardiopulmonary bypass. Three recipients died after transplantation secondary to acute right ventricular failure. In the surviving animals, NO led to significant improvements in pulmonary vascular resistance and vascular impedance, which occurred in association with significant increases in transpulmonary efficiency. No significant changes were observed in right and left ventricular preload recruitable stroke work after NO inhalation. CONCLUSIONS: These data suggest that NO may be an effective means to improve vascular impedance and pulmonary vascular efficiency after cardiac transplantation in the setting of recipient chronic pulmonary hypertension. PMID- 9205147 TI - Ex vivo adenoviral-mediated gene transfer to lung isografts during cold preservation. AB - BACKGROUND: Although whole-organ gene transfer has been reported in heart and liver transplant models, it has not been well characterized in lung grafts. The aim of this study was to determine the feasibility of ex vivo gene transfer to rat lung isografts during cold preservation using an adenoviral vector. METHODS: F344 rats, divided into four groups, underwent orthotopic left lung transplantation. In group I, lung grafts were flushed with adenovirus carrying the beta-galactosidase gene. After storage at 10 degrees C, grafts were implanted in recipient animals. Group II underwent the same procedure but graft storage was at 4 degrees C. Groups III (10 degrees C) and IV (4 degrees C) served as controls. On postoperative day 5, recipients were sacrificed, and native and transplanted lungs were examined. RESULTS: In group I, all animals showed successful, albeit patchy, gene expression. This occurred in 2 of 4 animals in group II, the other 2 showing no expression. Transduced cells were consistent morphologically with endothelial cells and pneumocytes. A minimal mononuclear inflammatory infiltrate was present. Control groups showed no transduction. CONCLUSIONS: It is feasible to perform ex vivo adenoviral-mediated gene transfer to rat lung isografts during cold preservation. PMID- 9205148 TI - Experimental bronchiolitis obliterans induced by in vivo HVJ-liposome-mediated endothelin-1 gene transfer. AB - BACKGROUND: Bronchiolitis obliterans (OB) is a lesion that results when injury to small conducting airways is repaired by a proliferation of fibrous granulation tissue. Bronchiolitis obliterans has emerged as a main cause of morbidity and mortality in the setting of lung and heart-lung transplantation. Endothelin-1 (ET 1), initially discovered as a vasoconstrictive peptide, has a mitogenic activity on vascular smooth cells and airway epithelial cells. Overproduction of endothelin has been reported in patients with OB or chronic rejection after lung transplantation. It is still undetermined whether locally overexpressed ET-1 has a potential impact in the pathogenesis of OB. METHODS: We locally overexpressed ET-1 using ultraviolet irradiation-inactivated hemagglutinating virus of Japan (HVJ)-liposome-mediated in vivo gene transfer. Plasmid DNA of prepro-ET-1 and high mobility group 1 protein were coencapsulated in liposomes, and were introduced into airway epithelial cells by HVJ-mediated membrane fusion. Control animals received instillation of HVJ-liposome with an empty expression cassette. To confirm the efficiency of transfection, HVJ liposome with beta-galactosidase gene was introduced. The expression of ET-1 and beta-galactosidase was assessed by immunohistochemistry. RESULTS: Bronchial epithelium alveolar cells and alveolar macrophage were stained blue (X-Gal) 1 week after in vivo gene transfer of beta-galactosidase gene, indicating beta-gal activity. In animals 1 to 2 weeks after in vivo transfection of prepro-ET-1 gene, hyperplastic connective tissue plaque was seen in the alveolar duct and small conducting airway, indicating histologically distinctive bronchiolitis obliterans. Strong ET-1-like immunoactivities were seen in the airway epithelial, hyperplastic connective tissue, and alveolar cells. No histopathologic changes were seen in the control animals. CONCLUSIONS: These results suggested that ET-1 may play an important role in the pathogenesis of OB. The effective pharmacologic antagonist or inhibitor may possibly control the progression of disease in patients of OB. PMID- 9205149 TI - Esophageal resection for cancer of the esophagus: long-term function and quality of life. AB - BACKGROUND: Information on function and quality of life of long-term survivors after esophageal resection for carcinoma is limited. METHODS: Between 1972 and 1990, 359 patients underwent esophagectomy for stage I or II esophageal carcinoma at Mayo Clinic. We evaluated long-term function and quality of life in 107 of these patients (81 men and 26 women) who survived 5 or more years. Median age at operation was 62 years (range, 30 to 81 years). The operation performed was an Ivor Lewis resection in 77 patients (72%), transhiatal esophagectomy in 14 (13%), extended esophagectomy in 4 (4%), thoracoabdominal esophagectomy in 4 (4%), and other in 8 (7%). Adenocarcinoma was present in 72 patients (67%), squamous cell carcinoma in 28 (26%), and other in 7 (7%). Thirty-four patients (32%) were in postsurgical stage I, 65 (61%) in stage IIA, and 8 (8%) in stage IIB. Median survival was 10.2 years (range, 5.0 to 23.2 years). Follow-up was complete for all patients. RESULTS: Gastroesophageal reflux was present in 64 patients (60%), symptoms of dumping in 53 (50%), and dysphagia to solid food in 27 (25%). Seventeen patients (16%) were asymptomatic. Factors affecting late functional outcome were analyzed. Patients who had a cervical anastomosis had significantly fewer reflux symptoms (p < 0.05). Dumping syndrome occurred more frequently in younger patients (p < 0.05) and women (p < 0.01). Quality of life was assessed separately by the Medical Outcomes Study 36-Item Short-Form Health Survey and compared with the national norm. Scores measuring physical functioning were decreased (p < 0.01). Scores measuring ability to work, social interaction, daily activities, emotional dysfunction, perception of health, and levels of energy were similar. Mental health scores were higher (p < 0.05). CONCLUSIONS: We conclude that long-term functional outcome after esophagectomy for esophageal carcinoma is affected by age, sex, and type of reconstruction. Quality of life as judged by the patients is similar to the national norm. PMID- 9205150 TI - Effect of bovine pericardial strips on air leak after stapled pulmonary resection. AB - BACKGROUND: Surgical procedures for emphysema have been proposed and in many settings resulted in significant improvement in dyspnea and function. The most prevalent surgical problem in all series is prolonged postoperative air leak. METHODS: One hundred twenty-three patients undergoing stapled thoracoscopic unilateral lung volume reduction operation were prospectively randomized to receive either no buttressing of their staple lines or buttressing of all staple lines with bovine pericardial strips. RESULTS: The two groups were comparable in preoperative risks and in the severity of their emphysema. Postoperative complications were identical in the two groups with respect to pneumonia, empyema, and wound infection; however, there was a significant difference in the duration of postoperative air leaks. Those having the pericardial strips used to buttress their staple lines had chest tubes removed 2.5 days sooner and were discharged from the hospital 2.8 days sooner as a result. The cost data revealed that because of the cost of the pericardial sleeves, the overall hospital charges were almost identical for the two groups ($22,108 bovine, $22,060 no bovine) in spite of the shortened hospital stay. CONCLUSIONS: The use of bovine pericardial sleeves to buttress the staple lines in thoracoscopic unilateral lung volume reduction operation results in a shorter duration of postoperative air leaks. Total hospital charges were comparable in the two groups as the 2.8 days saved in the hospital were offset by the cost of the pericardial sleeves. PMID- 9205152 TI - Emergency pulmonary embolectomy with percutaneous cardiopulmonary bypass. AB - BACKGROUND: The management of patients with acute pulmonary embolism remains difficult, particularly when cardiogenic shock is involved. The preoperative incidence of cardiac arrest compromises the results of emergency pulmonary embolectomy. In an attempt to reduce the operative mortality rate, we applied percutaneous cardiopulmonary bypass support to restore vital organ perfusion before the surgical intervention. METHODS: Percutaneous cardiopulmonary bypass support was preoperatively instituted in 3 patients with acute cardiopulmonary collapse caused by massive pulmonary embolism. In each patient, cardiac massage and endotracheal intubation were necessary due to loss of consciousness, hypotension, and severe cyanosis. Transesophageal echocardiography was performed to detect any evidence of thrombus in the main pulmonary artery, and each patient underwent the emergency pulmonary embolectomy using conventional cardiopulmonary bypass through a median sternotomy. RESULTS: Percutaneous cardiopulmonary bypass support immediately provided effective cardiopulmonary resuscitation. Transesophageal echocardiography clearly demonstrated any evidence of thrombus located in the pulmonary artery. Each patient was discharged from the hospital without any postoperative complication. CONCLUSIONS: The use of percutaneous cardiopulmonary bypass support immediately resuscitated and stabilized the cardiopulmonary function and allowed for successful emergency pulmonary embolectomy. In each patient, transesophageal echocardiography was useful for prompt and noninvasive diagnosis. PMID- 9205151 TI - Early and late airway complications after lung transplantation: incidence and management. AB - BACKGROUND: Airway anastomosis complications continue to be a source of morbidity for lung transplant recipients. METHODS: This study analyzes incidence, treatment, and follow-up of airway anastomotic complications occurring in 127 consecutive lung transplant airway anastomoses (77 single lung and 25 bilateral sequential lung). Complications were categorized as stenosis (11), granulation tissue (8), infection (7), bronchomalacia (5), or dehiscence (3). Follow-up after treatment ranged from 6 months to 4 years. RESULTS: Nineteen airway anastomosis complications (15.0%) occurred in 18 patients. Telescoping the airway anastomosis reduced the complication rate to 12 of 97 (12.4%), compared with 7 of 30 (23.3%) for omental wrapping, (p = 0.15). Complications developed in 13 of 77 single-lung airway anastomoses (16.9%) versus 6 of 50 bilateral sequential lung recipients (12.0%). Treatment consisted of stenting (9 airway anastomoses), bronchodilation (8), laser debridement (4), rigid bronchoscopic debridement (2), operative revision (2), and growth factor application (2). There was no difference in actuarial survival between patients with or without airway anastomosis complications (p = 1.0). CONCLUSIONS: Airway anastomosis complications can be successfully managed in the immediate or late postoperative period with good outcome up to 4 years after intervention. PMID- 9205153 TI - Pulmonary resection for lung trauma. AB - BACKGROUND: Pulmonary resection is rarely required for trauma, and its mortality is reportedly high. METHODS: A 10-year retrospective review of pulmonary resections for trauma was done. RESULTS: Of 2,455 patients with chest trauma, 183 (7.4%) underwent thoracotomy and 32 (1.3%) required pulmonary resection. Mean age was 28.4 years and mean injury severity score was 24.5. Mechanism of injury was stab wound in 14 patients, gunshot wound in 6, and blunt trauma in 12. Blunt trauma patients had a higher injury severity score (29.6) than penetrating trauma patients (21.4), but this was not significant (p < 0.07). Indications for thoracotomy were hemorrhage in 24 patients, airway disruption in 4, and other indications in 4. Operations consisted of wedge resection (19 patients), lobectomy (9), and pneumonectomy (4). Four (12.5%) patients (pneumonectomy, 2; lobectomy, 1; wedge, 1) died. Mortality for pneumonectomy was 50%, but this was not significantly higher than for lesser resections. Blunt trauma had a higher mortality (33%) than penetrating trauma (0%) (p < 0.02). Nonsurvivors had higher injury severity scores (44.2) than survivors (21.6) (p < 0.001). CONCLUSIONS: Pulmonary resection is infrequently required for lung injury. Overall mortality is lower than previously reported, but pneumonectomy has a high mortality. Blunt trauma has a higher mortality than penetrating trauma. Injury severity scores are higher for nonsurvivors than survivors; this shows the importance of associated injuries on outcome. PMID- 9205154 TI - Surgical management of ventricular tachycardia. AB - BACKGROUND: Ventricular tachyarrhythmias are the leading cause of death from coronary artery disease. A small percentage of these arrhythmias originate in chronically ischemic myocardium, rather than acutely ischemic myocardium, and can be refractory to medical management. Epicardial mapping and focal cryoablation of foci demonstrating early activation may provide definitive therapy when pharmacologic management fails. We report a series of 42 consecutive patients with refractory ventricular tachycardia (VT) who were treated with open epicardial mapping and focal cryoablation after pharmacologic management failed. METHODS: We retrospectively reviewed the records of patients who underwent surgical treatment of malignant VT. For patients not recently seen in the clinic, we conducted telephone interviews. At the time of operation, epicardial mapping was performed to locate foci of early electrical activation. These foci were then cryoablated, using 2-minute applications of liquid nitrogen-cooled probes. All patients underwent postoperative electrophysiologic studies to test for inducible VT. RESULTS: Of these 42 patients, 34 (81%) were male, 8 (19%) female. Average age was 62.9 +/- 10.6 years; ejection fraction, 0.20 (range, 0.04 to 0.50); and number of foci ablated, 2.1 +/- 1.1 (range, 1 to 6). At the time of cryoablation, all patients underwent additional procedures, including aneurysmectomy, coronary artery bypass, or valve replacement. The 30-day operative mortality was 9.5% (4 of 42). Of the 38 survivors, 36 (94.7%) were clinically free of VT; the remaining 2 had spontaneous or inducible VT. CONCLUSIONS: Open cryoablation of foci propagating VT appears to be safe and effective. It may be the most definitive treatment for malignant VT. PMID- 9205155 TI - Surgical management of renal cell carcinoma with inferior vena cava tumor thrombus. AB - BACKGROUND: The optimal management of patients with renal cell carcinoma with inferior vena cava tumor thrombus remains unresolved. Traditional approaches have included resection with or without the use of cardiopulmonary bypass. Chemotherapy has played a minor role except for biotherapeutic agents used for metastatic disease. METHODS: From January 1989 to January 1996, 37 patients with renal cell carcinoma and inferior vena cava tumor thrombus underwent surgical resection. The 27 men and 10 women had a median age of 57 years (range, 29 to 78 years). Thirty-six patients presented with symptoms; 21 had hematuria. Distant metastases were present in 12 patients. Tumor thrombi extended to the infrahepatic inferior vena cava (n = 16), the intrahepatic inferior vena cava (n = 16), the suprahepatic inferior vena cava (n = 3), and into the right atrium (n = 2). All tumors were resected by inferior vena cava isolation and, when necessary, extended hepatic mobilization and Pringle maneuver, with primary or patch closure of the vena cavotomy. Cardiopulmonary bypass was necessary in only 2 patients with intraatrial thrombus. RESULTS: Complications occurred in 11 patients, and 1 patient died 2 days postoperatively of a myocardial infarction (mortality, 2.7%). Twenty patients are alive; overall 2- and 5-year survival rates were 61.7% and 33.6%, respectively. For patients without lymph node or distant metastases (stage IIIa), 2- and 5-year survival rates were 74% and 45%, respectively. The presence of distant metastatic disease (stage IV) at the time of operation did not have a significant adverse effect on survival, as reflected by 2- and 5-year survival rates of 62.5% and 31.3%, respectively. Lymph node metastases (stage IIIc) adversely affected survival as there were no long-term survivors. CONCLUSIONS: Resection of an intracaval tumor thrombus arising from renal cell carcinoma can be performed safely and can result in prolonged survival even in the presence of metastatic disease. In our experience, extracorporeal circulatory support was required only when the tumor thrombus extended into the heart. PMID- 9205156 TI - Impact of retrograde cerebral perfusion on ascending aortic and arch aneurysm repair. AB - PURPOSE: The effect of retrograde cerebral perfusion on the incidence of stroke and death among patients undergoing repair of aneurysms of the ascending aorta and transverse arch was determined. MATERIALS AND METHODS: Between January 1991 and March 1995, 161 patients were operated on for aneurysms of the ascending aorta and transverse arch. Thirty-three of the patients (20%) had an aneurysm of the ascending aorta only and 128 (80%) had aneurysms of both the ascending aorta and the transverse arch. All the patients underwent cardiopulmonary bypass, profound hypothermia, and circulatory arrest, and 120 (74%) also underwent retrograde cerebral perfusion. Median pump time was 143 minutes (range, 21 to 461 minutes). Median circulatory arrest time was 42 minutes (range, 8 to 111 minutes), and median myocardial ischemic time was 71 minutes (range, 14 to 306 minutes). RESULTS: The overall 30-day mortality rate was 6% (9 patients) and the incidence of stroke was 4% (7 patients). The use of retrograde cerebral perfusion demonstrated a protective effect against stroke (3 of 120 patients, or 3%) compared with no retrograde cerebral perfusion (4 of 41 patients, or 9%; odds ratio, 0.24; confidence interval, 0.06 to 0.99; p < 0.049). This was most significant in patients more than 70 years of age; none of the 36 elderly patients who received retrograde cerebral perfusion had a stroke, compared with 3 of the 13 (23%) who did not (p < 0.003). Only pump time was associated with an increased risk of stroke (odds ratio, 1.01; 95% confidence interval, 1.00 to 1.02; p < 0.005). Pump time also was associated with increased mortality (odds ratio, 1.01; 95% confidence interval, 1.00 to 1.02; p < 0.008). CONCLUSION: Retrograde cerebral perfusion decreased the incidence of stroke in patients undergoing repair of aneurysms of the ascending aorta and transverse arch. PMID- 9205157 TI - Aortic valve replacement with patch enlargement of the aortic annulus. AB - BACKGROUND: Aortic annulus enlargement has long been advocated for the placement of valve prostheses larger than otherwise would have been possible. Little information exists, however, on the short- and long-term outcome of this surgical procedure. METHODS: We performed a retrospective review of 530 patients enrolled in a registry for patients who underwent aortic valve replacement using the Hancock II bioprosthesis and were followed up prospectively over the course of 11 years at a single institution. In an effort to avoid prosthetic valve-patient mismatch, the aortic annulus was enlarged in 98 patients (18%). Short- and long term outcome was analyzed. RESULTS: Enlargement of the aortic annulus during aortic valve replacement increased the operative mortality rate from 3.5% to 7.1%, but this difference did not reach statistical significance (p = 0.10). The long-term survival of patients who had annulus enlargement was similar to that of patients who did not. Because there were differences in the clinical profile of patients who had annulus enlargement and those who did not, a case-control study was carried out. This study showed similar long-term survival, freedom from valve related and cardiac death, and combined end points in the two groups of patients. CONCLUSION: Aortic annulus enlargement increased the operative mortality of aortic valve replacement. However, patients who underwent enlargement of a small aortic annulus had long-term survival and freedom from cardiac and valve-related death comparable to those of patients who received larger aortic prostheses. PMID- 9205158 TI - Risk factors and solutions for the development of neurobehavioral changes after coronary artery bypass grafting. AB - BACKGROUND: As operative mortality for coronary artery bypass grafting has decreased, greater attention has focused on neurobehavioral complications of coronary artery bypass grafting and cardiopulmonary bypass. METHODS: To assess risk factors and to evaluate changes in surgical technique, between 1991 and 1994 we evaluated 395 patients undergoing coronary artery bypass grafting with an 11 part neurobehavioral battery administered preoperatively and at 1 and 6 weeks postoperatively. Patients were instrumented with 5-MHz focused continuous-wave carotid Doppler transducers intraoperatively to estimate cerebral microembolism as an instantaneous perturbation of the velocity signal. Microembolism data were quantitated and compared with surgical technical maneuvers during operation and with neurobehavioral deficit (> or = 20% decline from preoperative performance on two or more neurobehavioral tests) postoperatively. These data and patient demographics were statistically analyzed (chi2, t test) and the results at 2 years (1991 and 1992; group A) were used to influence surgical technique in 1993 and 1994 (group B). RESULTS: Significantly associated with new neurobehavioral deficits were increasing patient age (p < 0.05), more than 100 emboli per case (p < 0.04), and palpable aortic plaque (p < 0.02). Group B patients had a significant decline in the neurobehavioral event rate (group A, 69%, 140/203; versus group B, 60%, 115/192; p < 0.05) of postoperative neurobehavioral deficits at 1 week and at 1 month (group A, 29%, 52/180; versus group B, 18%, 35/198; p < 0.01). The stroke rate was less than 2% in both groups (p = not significant). Modifications of surgical technique used in group B patients included increased use of single cross-clamp technique, increased venting of the left ventricle, and application of transesophageal and epiaortic ultrasound scanning to locate and avoid trauma to aortic atherosclerotic plaques. CONCLUSIONS: Neurobehavioral changes after coronary artery bypass grafting are common and associated with cerebral microembolization. Surgical technical maneuvers designed to reduce emboli production may improve neurobehavioral outcome. PMID- 9205159 TI - Evaluation of 7,000+ patients with two different routes of cardioplegia. AB - BACKGROUND: This study examined the efficacy and safety of retrograde cardioplegia in comparison with an antegrade/retrograde approach. METHODS: Between January 1, 1991, and December 31, 1995, 7,032 coronary artery bypass procedures, alone or in combination with valve replacement/repair, were performed using either retrograde cardioplegia (R) or an antegrade/retrograde (AR) approach. There were 4,224 patients in the R group and 2,808 in the AR group. These included elective, urgent, emergent/salvage, first operative, and redo cases. RESULTS: All preoperative, intraoperative, and postoperative variables listed in The Society of Thoracic Surgeons National Cardiac Surgery Database were used to compare the two groups using univariate analysis. The pump time was longer in the AR group, with fewer grafts per patient. The R group had higher predicted risk (3.2% versus 3.0%; p = 0.04), more postoperative atrial fibrillation (34% versus 31%; p = 0.006), and longer postoperative length of stay (8.8 versus 8.0 days; p < 0.001). Using The Society of Thoracic Surgeons National Cardiac Surgery Database predicted risk group model, a subgroup of 221 coronary artery bypass grafting patients in the retrograde (s-R) and 132 coronary artery bypass grafting patients in the antegrade/retrograde (s-AR) group fell into a greater incidence of predicted mortality group (> or = 10%). The s-R subgroup had more patients in New York Heart Association functional class IV. Univariate analysis revealed higher postoperative atrial fibrillation (51% versus 41%; p = 0.05) and longer postoperative length of stay (12.8 versus 10.8 days; p = 0.03) in the s-R subgroup versus the s-AR subgroup. CONCLUSIONS: The results appear to favor neither approach. Preoperatively, both retrograde groups (R and s-R) had higher preoperative predicted risk, but operative mortality or complications were not significantly increased when compared with the AR and s-AR groups. Retrograde cardioplegia alone was shown to be effective in the R and s-R groups, but atrial fibrillation developed in more patients, which could have contributed to longer length of stay in these groups. Antegrade/retrograde cardioplegia offers good immediate outcome but the delivery method can be cumbersome and confusing during the adjustments of flow clamps for antegrade/retrograde delivery and may contribute to prolonged pump times. From this retrospective, nonrandomized review, it appears that retrograde cardioplegia alone provides as good myocardial protection and safety as an antegrade/retrograde approach in either the low-risk or high-risk patient. PMID- 9205160 TI - Addition of alpha-ketoglutarate to blood cardioplegia improves cardioprotection. AB - BACKGROUND: We hypothesized that myocardial content of alpha-ketoglutarate (alpha KG), an intermediate of the Krebs cycle, can be critically low during heart operations, and that provision of alpha-KG could reduce metabolic abnormalities and lead to improved myocardial protection. METHODS: Twenty-four men aged 46 to 78 years who were undergoing heart operations participated in a prospective, controlled, randomized study. In 13 patients, an average of 28 g of alpha-KG was added to blood cardioplegia. Plasma creatine kinase isoenzyme MB and troponin T, and myocardial extraction of oxygen, substrates, and amino acids were measured. RESULTS: alpha-Ketoglutarate treatment was associated with lower creatine kinase isoenzyme MB (F = 39.6, df = 1.172, p < 0.001) and lower troponin (F = 12.9, df = 1.172, p < 0.001). The values at 4 hours were 31 +/- 2.4 microg/L versus 49 +/- 4.9 microg/L (creatine kinase isoenzyme MB) and 1.1 +/- 0.05 microg/L versus 2.0 +/- 0.34 microg/L (troponin T). Myocardial oxygen extraction was higher during alpha-KG cardioplegia (p < 0.01), but there were no significant differences in myocardial uptake or release of substrates or amino acids. Lactate release was observed in both groups during cardioplegia. Myocardial lactate release had ceased after 30 minutes of reperfusion in nearly half the alpha-KG-treated patients (6 of 13) but remained in all the control patients (11 of 11, p = 0.016). There were no other differences after 30 minutes of reperfusion. CONCLUSION: Provision of alpha-KG during blood cardioplegia improves myocardial protection in patients undergoing coronary operations. This may be linked to enhanced oxidation. PMID- 9205161 TI - Coronary artery bypass risk prediction using neural networks. AB - BACKGROUND: Neural networks are nonparametric, robust, pattern recognition techniques that can be used to model complex relationships. METHODS: The applicability of multilayer perceptron neural networks (MLP) to coronary artery bypass grafting risk prediction was assessed using The Society of Thoracic Surgeons database of 80,606 patients who underwent coronary artery bypass grafting in 1993. The results of traditional logistic regression and Bayesian analysis were compared with single-layer (no hidden layer), two-layer (one hidden layer), and three-layer (two hidden layer) MLP neural networks. These networks were trained using stochastic gradient descent with early stopping. All prediction models used the same variables and were evaluated by training on 40,480 patients and cross-validation testing on a separate group of 40,126 patients. Techniques were also developed to calculate effective odds ratios for MLP networks and to generate confidence intervals for MLP risk predictions using an auxiliary "confidence MLP." RESULTS: Receiver operating characteristic curve areas for predicting mortality were approximately 76% for all classifiers, including neural networks. Calibration (accuracy of posterior probability prediction) was slightly better with a two-member committee classifier that averaged the outputs of a MLP network and a logistic regression model. Unlike the individual methods, the committee classifier did not overestimate or underestimate risk for high-risk patients. CONCLUSIONS: A committee classifier combining the best neural network and logistic regression provided the best model calibration, but the receiver operating characteristic curve area was only 76% irrespective of which predictive model was used. PMID- 9205162 TI - Allograft aortic root replacement in prosthetic aortic valve endocarditis: a review of 32 patients. AB - BACKGROUND: This study was conducted to evaluate allograft aortic root replacement in the setting of complicated prosthetic valve endocarditis with extensive annular destruction. METHODS: From January 1990 through March 1996, 32 patients diagnosed with complicated prosthetic valve endocarditis underwent allograft root replacement. Mean age was 58.3 +/- 13.2 years; 23 patients were men. Mean preoperative New York Heart Association functional class was 3.4. Staphylococcus epidermidis (50%) and Enterococcus faecalis (19%) were the predominant causative microorganisms. Annular abscesses were found in 26 patients (81%), aortic-mitral discontinuity in 14 patients (43%), and left ventricular aortic discontinuity in 11 patients (34%). A cryopreserved allograft was used in 31 patients (97%) and a fresh antibiotic-treated allograft was used in 1 patient (3%). Mean aortic cross-clamp time was 150 +/- 29 minutes. Mean duration of the postoperative antibiotic treatment was 38.5 +/- 11.8 days. RESULTS: There were three operative deaths (9.4%); causes of death were multiorgan failure in 2 patients (6.2%) and low cardiac output in 1 patient (3.2%). Six patients (18%) had complete heart block (4 patients already before the operation), 3 patients (9.4%) had temporary respiratory insufficiency, and 1 patient (3.2%) needed temporary hemodialysis. Mean follow-up was 37.4 +/- 22.4 months. Two late deaths occurred: 1 patient had recurrent endocarditis, leading to a false aneurysm, and died at reoperation; another patient died of lung cancer. Actuarial 5-year survival was 87.3% (70% confidence interval, 76.8% to 97.8%); actuarial 5-year freedom from recurrent endocarditis was 96.5% (70% confidence interval, 90.0% to 100%). CONCLUSIONS: Allograft aortic root replacement is a valuable technique in the complex setting of prosthetic valve endocarditis with involvement of the periannular region. Mortality and morbidity are low. PMID- 9205163 TI - Results after surgical repair of Ebstein's anomaly. AB - BACKGROUND: Ebstein's anomaly of the tricuspid valve is a complex malformation. Various operations have been undertaken with varying results. Because valve replacement yielded poor results, surgical treatment has focused on valvuloplasties. METHODS: Between April 1974 and February 1995, 60 patients with Ebstein's anomaly underwent surgical repair. Age ranged from 5 months to 54 years. In 56 patients (93.3%), tricuspid valvuloplasty was feasible, mainly by creating a monocusp valve with the single-stitch technique. The other 4 patients had valve replacement with a bioprosthesis. Six reoperations were necessary (10.0%): four valve replacements and two repeat valvuloplasties. RESULTS: There were two hospital deaths (3.3%) and a late mortality rate of 10.0% (6 patients). Forty-nine (94.2%) of 52 survivors were followed for 5 months to 18.6 years (median follow-up, 5.0 years; mean follow-up, 6.9 years). The actuarial survival rate (Kaplan-Meier) was 96.5% +/- 2.4% at 1 year and 83.3% +/- 5.6% at 18 years. At follow-up evaluation, nearly all patients showed substantial improvement (93.9% were in functional class I or II) compared with their preoperative status. Doppler echocardiographic studies demonstrated good tricuspid valve function in most patients. CONCLUSIONS: Valvuloplasty using the single-stitch technique is a rewarding operation. It yields good long-term results with substantial improvement in functional performance and clinical status. PMID- 9205164 TI - Predicting feasibility of biventricular repair of right-dominant unbalanced atrioventricular canal. AB - BACKGROUND: In right-dominant unbalanced atrioventricular (AV) canal, there are no criteria to judge adequacy of the left ventricle for biventricular repair. The purpose of this study was to test the hypothesis that right ventricular volume overload in this condition results in right-to-left septal bowing and contributes to the appearance of a small left ventricle. METHODS: Five consecutive neonates and young infants (age range, 23 days to 5 months; median age, 3 months) with right-dominant unbalanced complete AV canal underwent biventricular repair. Preoperative and postoperative echocardiographic measurements of left (LV) and right ventricular size and AV valve component size were made. Potential LV volume was assessed preoperatively using a theoretic model that assumed a normalization of septal bowing. RESULTS: There was no perioperative mortality; 1 patient died 71 days postoperatively of problems related to the left AV valve. Preoperatively, all patients had severe LV hypoplasia, with a mean end-diastolic indexed true LV volume of 14.8 +/- 9.1 mL/m2, indexed potential LV volume of 32.0 +/- 18.8 mL/m2, left AV valve to total AV valve ratio of 0.30 +/- 0.06, and LV to right ventricular long-dimension ratio of 0.65 +/- 0.1. Postoperatively, all patients had indexed true LV volumes greater than 30 mL/m2 (mean volume, 35.6 +/- 3.9 mL/m2), and the left AV valve to total AV valve ratio and the LV to right ventricular long-dimension ratio increased to 0.42 +/- 0.03 and 0.88 +/- 0.11, respectively. Both preoperative potential and true LV volumes correlated well with postoperative true LV volumes: r = 0.90 (p = 0.040) and r = 0.93 (p = 0.023), respectively. Increases in LV length and left AV annulus size indicated contributions of volume loading and surgical patching to the right of the ventricular crest to the increase in LV size. CONCLUSIONS: In our small series, preoperative indexed potential LV volume of 15 mL/m2 or greater (present in all patients) allowed biventricular repair of right-dominant unbalanced AV canal. Any previous criteria for LV hypoplasia in this condition need to be reconsidered. This study also has implications for other right-sided volume-loaded lesions in which the left ventricle initially is judged to be hypoplastic but in which biventricular repair may be feasible. PMID- 9205165 TI - Intravenous phenylephrine preconditioning of cardiac grafts from non-heart beating donors. AB - BACKGROUND: Hypoxia and warm ischemia produce severe injury to cardiac grafts harvested from non-heartbeating donors. To potentially improve recovery of such grafts, we studied the effects of intravenous phenylephrine preconditioning. METHODS: Thirty-seven blood-perfused rabbit hearts were studied. Three groups of non-heart-beating donors underwent intravenous treatment with phenylephrine at 12.5 (n = 8), 25 (n = 7), or 50 microg/kg (n = 7) before initiation of apnea. Non heart-beating controls (n = 8) received saline vehicle. Hypoxic cardiac arrest occurred after 6 to 12 minutes of apnea, followed by 20 minutes of warm in vivo ischemia. A 45-minute period of ex vivo reperfusion ensued. Nonischemic controls (n = 7) were perfused without antecedent hypoxia or ischemia. RESULTS: Phenylephrine 25 microg/kg significantly delayed the onset of hypoxic cardiac arrest compared with saline controls (9.6 +/- 0.5 versus 7.7 +/- 0.4 minutes; p = 0.00001), yet improved recovery of left ventricular developed pressure compared with saline controls (57.1 +/- 5.3 versus 41.0 +/- 3.4 mm Hg; p = 0.04). Phenylephrine 25 microg/kg also yielded a trend toward less myocardial edema than saline vehicle (p = 0.09). CONCLUSIONS: Functional recovery of nonbeating cardiac grafts is improved by preconditioning. We provide evidence that the myocardium can be preconditioned with phenylephrine against hypoxic cardiac arrest. PMID- 9205166 TI - Expression of immediate early genes after cardioplegic arrest and reperfusion. AB - BACKGROUND: Induction of protooncogenes such as c-fos, c-jun, and EGR-1 has been implicated in cellular growth and differentiation. Heat-shock proteins (HSPs) such as hsp 70 may mediate resistance to ischemia after heat shock and ischemic preconditioning. The effects of cardioplegia on the regulation of these immediate early genes are unclear. METHODS: Isolated rat hearts were subjected to different cold (5 degrees C) or normothermic (35 degrees C) cardioplegic solutions and reperfused with normothermic Krebs-Henseleit buffer. Right atrial biopsy specimens from patients undergoing coronary artery bypass grafting with cold cardioplegic arrest were taken before and after cardiopulmonary bypass. Analysis of immediate early gene messenger RNAs was performed using Northern blots. Related proteins were localized by immunohistochemistry. RESULTS: In rat hearts, cold cardioplegia for 40 minutes with Bretschneider or St. Thomas' II solution followed by 40 minutes' reperfusion resulted in a significant increase in left ventricular c-fos, EGR-1, and c-jun messenger RNA levels (4.0-, 3.1-, and 3.0 fold, respectively, with Bretschneider solution and 3.7-, 2.8-, and 2.1-fold, respectively, with St. Thomas' II solution) compared with control hearts perfused at 35 degrees C with Krebs-Henseleit buffer. Normothermic cardioplegia with St. Thomas' II solution was without effect, whereas sequential perfusion with Krebs Henseleit buffer at 5 degrees C and 35 degrees C resulted in a similar increase in protooncogene messenger RNA levels. Only cold Bretschneider solution was related to a 5.2-fold induction of hsp 70 messenger RNA levels. Likewise, rat atrial tissues and samples from patients after cardiopulmonary bypass displayed a significant induction of these immediate early genes. Monoclonal antibodies against c-FOS and HSP 70 proteins stained nuclei and perinuclear spaces of endothelial cells and cardiac myocytes. CONCLUSIONS: Cold cardioplegic arrest and normothermic reperfusion are potent triggers for immediate early gene induction. Hypothermia emerged as the prime stimulus for the examined protooncogenes. In contrast, hsp 70 induction was dependent on the cardioplegic solution. PMID- 9205167 TI - Bidirectional cavopulmonary shunt in patients with anomalies of systemic and pulmonary venous drainage. AB - BACKGROUND: Bidirectional cavopulmonary shunt and Fontan repair are now commonly performed in patients with a variety of forms of complex single ventricle, including those with anomalies of systemic or pulmonary venous return. These anomalies are ideally dealt with during bidirectional cavopulmonary shunt, thereby minimizing the complexity of the eventual Fontan procedure. METHODS: Between March 1990 and December 1995, 36 patients with anomalous systemic or pulmonary venous drainage underwent bidirectional cavopulmonary shunt. A combination of anomalous systemic and pulmonary venous drainage was present in 12 patients, whereas 19 patients had anomalous drainage only from the systemic circulation and 5 patients had isolated anomalies of pulmonary venous return. Visceral heterotaxy syndrome was diagnosed in 18 patients. The median age at operation was 11 months, and bidirectional cavopulmonary shunt was the first surgical procedure performed in 10 of these patients. Techniques of repair are described. RESULTS: There were two early deaths and one bidirectional cavopulmonary shunt was taken down, for mortality and failure rates not significantly different than those for all patients undergoing bidirectional cavopulmonary shunt during this time period (n = 117). At a mean follow-up of 19.9 months, there have been three late deaths and 11 patients have undergone Fontan completion. Actuarial survival was 87% at 1 year and 81% at 3 years. Among all patients undergoing bidirectional cavopulmonary shunt during this time period, neither heterotaxy syndrome nor anomalies of systemic or pulmonary venous return were significantly associated with decreased survival or poor outcome. CONCLUSIONS: Bidirectional cavopulmonary shunt can be performed in patients with anomalous systemic or pulmonary venous drainage, including those with visceral heterotaxy syndrome, with morbidity and mortality rates that do not differ significantly from those achieved in all patients undergoing bidirectional cavopulmonary shunt. In this report, we describe our experience with this group of patients, primarily focusing on outcomes and technical issues that pertain to the use of bidirectional cavopulmonary shunt as a preparatory procedure for the extracardiac conduit Fontan operation. PMID- 9205168 TI - Cardiac operations in patients 90 years of age and older. AB - BACKGROUND: Growth of the elderly population worldwide, and specifically in the United States, will continue to accelerate and will have a profound impact on the cost and delivery of health care resources in the future. A medical strategy that allows the elderly to live independently is essential to most cost-effective use of our resources. The question remains as to what will be the future of surgical therapy for this increasing population. METHODS: We retrospectively studied the cases of 30 consecutive nonagenarians (mean age, 92.3 +/- 1.8 years) who underwent a cardiac operation within a 9-year period. All patients were in New York Heart Association class III or IV and underwent operation urgently or emergently. RESULTS: The 30-day mortality rate was 10%, and the actuarial survival rates were 81% +/- 8% and 75% +/- 9% at 1 year and 2 years, respectively. Seventy-eight percent of survivors were in New York Heart Association class I or II within 2 years after operation and had an improved quality of life. The cost of providing care in this age group was 24% higher than in octogenarians. CONCLUSIONS: Advanced age in and of itself (>90 years) should not be a contraindication to an open-heart operation, although morbidity, mortality, and cost may be higher. However, selective criteria identifying risks and benefits for individual patients should be applied. The aging of our population will have a profound impact on the cost and delivery of health care resources in the future. This issue must be addressed in the current debate on the provision of expensive procedures under a realigned national health-care system. PMID- 9205169 TI - Effect of surgical reconstruction on flow profiles in the aorta using magnetic resonance blood tagging. AB - BACKGROUND: The aorta that has undergone an aorta-pulmonary artery anastomosis may not exhibit the same velocity profile as the nonreconstructed aorta, whose velocity profile is thought to be uniform across the vessel diameter (plug flow). This may have an impact on fluid dynamics and will alter Doppler flow calculations. Our objective was to determine the impact of surgical reconstruction on the velocity and flow profiles of the reconstructed ascending and descending aorta. METHODS: Using a magnetic resonance imaging tagging technique that labels flowing blood (bolus tagging), we studied 22 patients (mean age, 8.6 +/- 4.7 years) who had had a Fontan procedure. A cine sequence labeled the blood and acquired the image after 20 ms in the middle of the ascending aorta and behind the left atrium in the descending aorta. The repetition time was 50 ms. RESULTS: The reconstructed ascending aorta displayed a velocity profile skewed anteriorly, whereas in the nonreconstructed aorta, the velocity profile was flat. Reconstructed aortas also displayed flows that were higher anteriorly, took a longer time to reach maximum velocity, and were less like "plug" flow than the nonreconstructed aorta. The descending aorta, regardless of whether aortic reconstruction was present, displayed velocity profiles (at various phases of systole) skewed posteriorly. CONCLUSIONS: The reconstructed aorta displays disturbed flow, and the velocities across the ascending aortic diameter are more varied than those in aortas without reconstruction and are skewed anteriorly. The descending aortic velocity profile in children is skewed posteriorly, regardless of whether aortic reconstruction is present. This information may help design and build a "better" aortic reconstruction. PMID- 9205170 TI - Heparin-bonded circuits decrease myocardial ischemic damage: an experimental study. AB - BACKGROUND: Heparin-bonded cardiopulmonary bypass circuits reduce complement activation, but their effect on myocardial function is unknown. This study was undertaken to determine whether heparin-bonded circuits reduce myocardial damage during acute surgical revascularization. METHODS: In 16 pigs, the second and third diagonal vessels were occluded with snares for 90 minutes followed by 45 minutes of cardioplegic arrest and 180 minutes of reperfusion with the snares released. During the period of coronary occlusion, all animals were placed on percutaneous bypass followed by standard cardiopulmonary bypass during the periods of cardioplegic arrest and reperfusion. In 8 pigs, heparin-bonded circuits were used, whereas 8 other pigs received nonbonded circuits. RESULTS: Animals treated with heparin-bonded circuits had the best preservation of wall motion scores (3.5 +/- 0.3 versus 2.3 +/- 0.2; 4 = normal to -1 = dyskinesis; p < 0.05), least tissue acidosis (change in pH = -0.31 +/- 0.02 versus -0.64 +/- 0.08; p < 0.05), smallest increase in lung H2O (1.7% +/- 0.7% versus 6.1% +/- .5%; p < 0.05), and the lowest area of necrosis/area of risk (20.3% +/- 2.2% versus 40.4% +/- 1.6%; p < 0.05). CONCLUSIONS: We conclude that heparin-bonded circuits significantly decrease myocardial ischemic damage during acute surgical revascularization. PMID- 9205171 TI - Effects of wrapping tightness on acute cardiac function in dynamic cardiomyoplasty. AB - BACKGROUND: It has not been clarified how tightly the heart should be wrapped for maximal augmentation of cardiac function in cardiomyoplasty. METHODS: Hearts in acute failure induced by propranolol were wrapped with the left latissimus dorsi muscle, loosely (loose CMP), moderately (moderate CMP), and tightly (tight CMP) in each of 5 pigs. To measure the pressure between the latissimus dorsi muscle and the left ventricle (LV), a Millar pressure catheter with a latex balloon was placed on the anterior wall of the LV. Left ventricular wall tension was calculated according to Laplace's law, using the difference between the LV pressure and the balloon pressure. RESULTS: In the loose CMP, moderate CMP, and tight CMP groups, the mean balloon pressures during unassisted beats were 8.2, 10.4, and 13.2 mm Hg, respectively. During unassisted beats, the mean LV wall tension values were 38,683, 29,938 (p < 0.05 versus loose CMP), and 26,652 (p < 0.05 versus loose CMP) dynes/cm, respectively, the peak LV pressures were 76.8, 73.8, and 65 (p < 0.05 versus loose CMP) mm Hg, respectively, and the stroke volumes were 12.8, 9.2, and 8.5 (p < 0.05 versus loose CMP) mL, respectively. During assisted beats, the mean LV wall tension values were 20,059, 11,290, and 7,893 (p < 0.05 versus loose CMP) dynes/cm, respectively, the peak LV pressures were 94.1, 98.1, and 92.0 mm Hg, respectively, and the stroke volumes were 13.8, 11.6, and 9.4 (p < 0.05 versus loose CMP) mL, respectively. CONCLUSIONS: During unassisted beats, tight CMP (13 mm Hg) had the advantage of diminishing LV wall tension, but the disadvantage of diminishing LV pressure and stroke volume, compared with loose CMP (8 mm Hg). Moderate CMP (10 mm Hg), however, had the advantage of diminishing LV wall tension without a decrease in LV pressure and stroke volume. PMID- 9205173 TI - Is there an advantage to repairing infected mitral valves? AB - BACKGROUND: The therapy for native mitral valve endocarditis is in evolution. Antibiotics have significantly improved survival rates, but patients with complications of endocarditis may require surgical treatment. METHODS: Between January 1985 and December 1995, 146 patients underwent surgical therapy (repair or replacement) for native mitral valve endocarditis. All patients had documented bacterial endocarditis. Univariate and multivariate analyses were performed to determine predictors of hospital death, long-term event-free survival, and probability of repair. Patients were evaluated in three groups: all patients, patients with acute endocarditis, and patients with chronic endocarditis. RESULTS: There were ten hospital deaths (6.8%). Patients undergoing repair had a lower hospital mortality rate (p = 0.008) then those having replacement. Event free survival was improved after mitral valve repair in the overall group (p = 0.02) and in the group with healed (chronic) endocarditis (p = 0.05). Although the acute endocarditis group demonstrated an improved event-free survival rate after mitral valve repair versus replacement (74% versus 20% at 6 years), this did not reach statistical significance. CONCLUSIONS: We conclude that mitral valve repair is preferable to mitral valve replacement when possible, in patients with complications of endocarditis, as repair results in a lower hospital mortality and an improved long-term survival. PMID- 9205172 TI - Clinical outcome and left ventricular function after pulmonary autograft implantation in children. AB - BACKGROUND: Aortic root replacement with a pulmonary autograft is an alternative treatment for children with aortic valve or root disease, or both. METHODS: Twenty-six patients (18 boys and 8 girls) with a mean age of 10.9 years (range, 0.3 to 16.9 years) underwent this procedures in a 7-year period. The mean follow up period was 3.2 years (range, 0.2 to 7.5 years). RESULTS: During follow-up 3 patients died and one autograft was replaced with a mechanical valve. The actuarial survival and actuarial event-free survival rates were 87% and 79%, respectively, at both 5 and 7 years. None of the surviving patients had complaints, and all have done well and are living normal lives. Electrocardiographic signs of myocardial ischemia and left ventricular hypertrophy were not present. Echocardiography showed autograft valve regurgitation to be absent or trivial (n = 17) or mild (n = 5). Stenosis was not present. Increasing autograft annulus diameters were noted during follow-up, but this was not related to the severity of autograft regurgitation. Left ventricular dimensions and function were within normal limits later than 1 year after the operation. Only 2 patients had a moderate pulmonary stenosis without right ventricular hypertrophy. CONCLUSIONS: The surgical results, clinical outcome, valve function, and left ventricular function in our patients have been good. This procedure is recommended as a method of aortic valve replacement in children. PMID- 9205174 TI - The brain uses mostly dissolved oxygen during profoundly hypothermic cardiopulmonary bypass. AB - BACKGROUND: During profoundly hypothermic cardiopulmonary bypass, cerebral venous oxygen saturation increases (eg, to 98% at 15 degrees C). We reanalyzed results of clinical studies to learn why. METHODS: One hundred sixty-eight cerebral oxygen transport measurements were available from 96 infants and children undergoing profoundly hypothermic cardiopulmonary bypass during repair of congenital heart defects. RESULTS: Dissolved oxygen accounted for 2% to 17% of arterial oxygen content, depending on the arterial oxygen partial pressure and hemoglobin concentration. The fraction of the cerebral metabolic rate for oxygen obtained from dissolved oxygen depended on pump flow, temperature, hemoglobin concentration, and arterial oxygen partial pressure (all p < 10(-3)). For "full flow" cardiopulmonary bypass, temperatures less than 18 degrees C, and arterial oxygen partial pressure measurements more than 180 mm Hg, the mean +/- standard deviation of the fraction of cerebral metabolic rate for oxygen obtained from dissolved oxygen equaled 77% +/- 19%. CONCLUSIONS: Dissolved oxygen satisfies most of the brain's oxygen requirements during profound hypothermic cardiopulmonary bypass. This result reflects four properties of profound hypothermic cardiopulmonary bypass: (1) increases in hemoglobin's oxygen affinity due to profound hypothermia (which impairs oxygen transfer from hemoglobin to cerebral tissue), (2) use of hemodilution, (3) use of high arterial oxygen partial pressure, and (4) low cerebral metabolic rate of oxygen. PMID- 9205175 TI - Silicone-coated polypropylene hollow-fiber oxygenator: experimental evaluation and preliminary clinical use. AB - BACKGROUND: A membrane oxygenator consisting of a microporous polypropylene hollow fiber with a 0.2-microm ultrathin silicone layer (cyclosiloxane) was developed. Animal experimental and preliminary clinical studies evaluated its reliability in bypass procedures. METHODS: Five 24-hour venoarterial bypass periods were conducted on dogs using the oxygenator (group A). In 5 controls, bypass periods were conducted using the same oxygenator without silicone coating (group B). As a preliminary clinical study, 14 patients underwent cardiopulmonary bypass with the silicone-coated oxygenator. RESULTS: Eight to 16 hours (mean, 12.2 hours) after initiation of bypass, plasma leakage occurred in all group B animals, but none in group A. The O2 and CO2 transfer rates after 24 hours in group A were significantly higher than at termination of bypass in group B (p < 0.005 and p < 0.03, respectively). Scanning electron microscopy of silicone coated fibers after 24 hours of bypass revealed no damage to the silicone coating of the polypropylene hollow fibers. In the clinical study, the oxygenator showed good gas transfer, acceptable pressure loss, low hemolysis, and good durability. CONCLUSIONS: This oxygenator is more durable and offers greater gas transfer capabilities than the previous generation of oxygenators. PMID- 9205176 TI - Active native valve endocarditis: determinants of operative death and late mortality. AB - BACKGROUND: In this report, we reviewed 247 patients who underwent operation by our team for active native valve endocarditis between January 1979 and December 1993. METHODS: There were 201 male and 46 female patients (mean age, 45.4 +/- 6 years). The aortic valve was involved in 163 cases, the mitral valve in 36 cases, both mitral and aortic valves in 44 cases, and the tricuspid valve alone in 4 cases. The most common microorganisms were streptococci. Univariate Pearson (chi2 test) and multivariate (stepwise logistic regression [BMDPLR]) analyses were used to identify significant predictors of operative mortality, reoperation, and recurrent endocarditis. Cox proportional hazards regression model was used to study late survival. RESULTS: Operative mortality was 7.6% (n = 19). Increased age, cardiogenic shock at the time of operation, insidious illness, and greater thoracic ratio (>0.5) were the predominant risk factors; the length of antibiotic therapy appeared to have no influence. Two hundred thirteen patients were followed up. Median follow-up time was 6 years (range, 2 to 19 years). Overall survival rate (operative mortality excluded) was 71.3% +/- 3.8% at 9 years. Increased age, preoperative neurologic complications, cardiogenic shock at the time of operation, shorter duration of the illness, insidious illness before the operation, and mitral valve endocarditis were the predominant risk factors for late mortality. The probability of freedom from reoperation (operative mortality included) was 73.3% +/- 4.2% at 8 years; risk factors were younger age and aortic valve endocarditis. The rate of prosthetic valve endocarditis was 7%. No significant risk factor was found. CONCLUSIONS: Increased age, insidious illness, and hemodynamic failure are the main risk factors for operative mortality. Long term survival is good except for patients with preoperative neurologic complications and mitral valve endocarditis. PMID- 9205177 TI - Initial experience using an intraluminal shunt during revascularization of the beating heart. AB - BACKGROUND: For decades, surgeons have relied on extracorporeal circulation and induced cardiac asystole to provide a bloodless, motionless field in which to construct coronary bypass grafts. However, the technique of coronary grafting without heart-lung support is now being revitalized. The current resurgence of off-pump coronary artery bypass grafting and the advent of less invasive incisions make it imperative that technical advances be applied to maximize the safety of these procedures. METHODS: This report describes an inexpensive intraluminal shunt that maintains coronary perfusion, prevents ischemia, reduces backbleeding, and molds the suture line to prevent accidental missuturing of the posterior coronary wall. RESULTS: In 63 patients, saphenous grafts were placed to the left anterior descending (49), diagonal (9), and right coronary artery (27) without extracorporeal circulation using an intraluminal shunt. There were no deaths (0% mortality) and one perioperative infarction (1.5%). Complication and graft patency rates were comparable with those obtained by conventional techniques. CONCLUSIONS: Temporary intraluminal shunting greatly facilitates the surgeons' operative environment by permitting safe and precise construction of coronary artery grafts on the beating heart in a bloodless field. Intraluminal shunting may have future implications on the ability to perform safe and reproducible grafting on the beating heart through minimally invasive or endoscopic approaches. PMID- 9205178 TI - Closed-chest cardiopulmonary bypass and cardioplegia: basis for less invasive cardiac surgery. AB - BACKGROUND: We developed a method of closed-chest cardiopulmonary bypass to arrest and protect the heart with cardioplegic solution. This method was used in 54 dogs and the results were retrospectively analyzed. METHODS: Bypass cannulas were placed in the right femoral vessels. A balloon occlusion catheter was passed via the left femoral artery and positioned in the ascending aorta. A pulmonary artery vent was placed via the jugular vein. In 17 of the dogs retrograde cardioplegia was provided with a percutaneous coronary sinus catheter. RESULTS: Cardiopulmonary bypass time was 111 +/- 27 minutes (mean +/- standard deviation) and cardiac arrest time was 66 +/- 21 minutes. Preoperative cardiac outputs were 2.9 +/- 0.70 L/min and postoperative outputs were 2.9 +/- 0.65 L/min (p = not significant). Twenty-one-French and 23F femoral arterial cannulas that allowed coaxial placement of the ascending aortic balloon catheter were tested in 3 male calves. Line pressures were higher, but not clinically limiting, with the balloon catheter placed coaxially. CONCLUSIONS: Adequate cardiopulmonary bypass and cardioplegia can be achieved in the dog without opening the chest, facilitating less invasive cardiac operations. A human clinical trial is in progress. PMID- 9205179 TI - Extension of the "elephant trunk" technique in complex aortic pathology: the "bidirectional" option. AB - BACKGROUND: The "elephant trunk" technique, using a free-floating vascular prosthesis, was originally described to facilitate a subsequent operation on the downstream aorta. We developed an additional refinement of this technique, called the "bidirectional elephant trunk." This option may represent an interesting tool in more complex aortic operations, especially when the descending aorta has to be replaced first in patients with concomitant pathology of the ascending aorta or of the aortic arch. METHODS: The initial operation is performed through a left thoracotomy. The proximal elephant trunk is created by invaginating the future aortic arch graft into the descending aortic graft. The proximal anastomosis between the doubled graft and the proximal descending aorta is performed first. During construction of the distal anastomosis, a distal elephant trunk may be inserted likewise. If the aortic arch and ascending aorta have to be replaced later, this second step is performed through a median sternotomy. The free floating arch graft is pulled out of the proximal descending aorta with a nerve hook, unfolded, and used for total arch replacement. RESULTS: This technique was used successfully in 3 patients without mortality. No major complications were observed excepted persistent hoarseness in a patient with preoperative paresis of the recurrent nerve. No perfusion problems due to the unfolding of the free floating graft occurred during the second operation. CONCLUSIONS: The bidirectional elephant trunk technique is an interesting option that may be suitable for patients presenting with a complex pathology of the whole thoracic aorta when the descending segment has to be replaced first. PMID- 9205180 TI - Persistence of mammary artery branches and blood supply to the left anterior descending artery. AB - BACKGROUND: Partial harvesting of the left internal mammary artery (LIMA) is a widespread technique used during minimally invasive coronary operations performed through a left anterior small thoracotomy. The influence of persisting LIMA branches was investigated to evaluate their effect on the blood flow of the left anterior descending artery. METHODS: Thirty patients, 15 with totally (group A) and 15 with partially (group B) harvested LIMAs, were evaluated. All the patients underwent postoperative angiography, during which a flow map of the LIMA was performed. The average peak velocity and the diastolic-to-systolic peak velocity ratio were recorded. The LIMA graft flow pattern was recorded in the proximal and distal thirds of the artery. Intramammary adenosine (12 to 14 microg) was injected and the average peak velocities before and after injection were calculated. RESULTS: The average peak velocity was similar in both groups in the proximal and distal thirds of the LIMA (25 +/- 7 and 26 +/- 5 cm/sec, respectively, in group A versus 27 +/- 5 and 25 +/- 5 cm/sec, respectively in group B; p = NS). The diastolic-to-systolic peak velocity ratio was similar proximally (0.78 +/- 0.3 in group A versus 0.69 +/- 0.3 cm/s in group B; p = NS), but not distally (1.72 +/- 0.1 in group A versus 0.97 +/- 0.3 in group B; p < 0.0005). The LIMA graft flow reserve was similar both proximally and distally (2.6 +/- 0.6 and 2.5 +/- 0.3 cm/s, respectively, in group A versus 2.6 +/- 0.5 and 2.6 +/- 0.3 cm/s, respectively, in group B; p = NS). CONCLUSIONS: The persistence of LIMA branches does not influence the blood flow of the left anterior descending artery after acute adenosine-induced myocardial hyperemia. If a left anterior small thoracotomy is used in left anterior descending artery direct revascularization, complete LIMA harvesting is not mandatory and depends on the personal preference of the surgeon. PMID- 9205181 TI - Embolization of IMA side branch for post-CABG ischemia. AB - The existence of a chest wall "steal" of blood away from the myocardium through patent internal mammary artery branches has been hypothesized as a cause of recurrent angina pectoris after coronary artery bypass grafting. Although some authors believe that such a steal is physiologically impossible because coronary flow occurs in diastole and chest wall flow in systole, we recently documented ischemia in the left anterior descending coronary artery distribution before embolization of a large left internal mammary artery first intercostal branch that had been left intact at the time of operation. After embolization of the branch, clinical and objective evidence of ischemia resolved. PMID- 9205182 TI - Osteogenic sarcoma of the left atrium. AB - A primary cardiac osteogenic sarcoma is described in a patient presenting with a left atrial obstructive lesion. Wide surgical excision was possible with left atrial reconstruction and mitral valve replacement. The patient survived the operation and was symptom free for 1 year, finally dying at 18 months of cerebral metastases. PMID- 9205183 TI - Torn ascending aorta missed by transesophageal echocardiography. AB - Transesophageal echocardiography has become a commonly used screening tool for traumatic tears of the descending aorta. The role of transesophageal echocardiography for ascending aortic tears is not yet well-defined. We report an ascending aortic tear imaged by aortography but missed on transesophageal echocardiography. PMID- 9205184 TI - Marfanoid aneurysm in donor aorta after transplantation. AB - A case is reported of dissecting aneurysm of the donor ascending aorta and root 4 years after orthotopic cardiac transplantation. The pathology raises the possibility of Marfan's syndrome in the donor. PMID- 9205185 TI - Chondrosarcoma originating from the trachea. AB - We report a tracheal chondrosarcoma in a 54-year-old man, treated with neodymium:yttrium-aluminum garnet laser vaporization via fiberoptic bronchoscopy followed by surgical resection. Chondrosarcomas of the trachea are extremely rare tumors. To our knowledge, there are 9 cases of chondrosarcoma of the trachea reported in the English-language literature, to which we add the tenth. PMID- 9205186 TI - Recanalization of the left atrial appendage demonstrated by transesophageal echocardiography. AB - Closure of the fibrillating left atrial appendage has been recommended during mitral valve operations to help prevent thrombus formation and systemic embolization postoperatively. We report recanalization of the appendage orifice in 6 patients after surgical closure by pursestring suturing at the time of mitral valve replacement. Transesophageal echocardiography demonstrated disruption of the closure line and partial recanalization of the sutured orifice with relatively high velocity flow between the left atrial body and the appendage. PMID- 9205187 TI - Minimally invasive axillary-coronary artery bypass. AB - Axillary artery-to-coronary artery bypass using reversed saphenous vein provides a simple method of applying the minimally invasive coronary bypass grafting procedure when the internal thoracic artery is not an adequate conduit. Although this may allow extended use of the minimally invasive coronary bypass procedure, the long-term patency of this technique is unknown. PMID- 9205188 TI - Treatment of refractory, nonmalignant hydrothorax with a pleurovenous shunt. AB - We present a case of long-term successful application of pleurovenous shunting for the management of pleural effusion. Intractable symptomatic hydrothorax developed as a result of transdiaphragmatic migration of hepatic ascites. After failure of traditional treatment by mechanical pleurodesis, a pleurovenous shunt was inserted. After 1 year of follow-up, the effusion is well controlled, and the shunt remains patent. PMID- 9205189 TI - Successful repair of primary concomitant aortobronchial and aortoesophageal fistulas. AB - Combined aortobronchial and aortoesophageal fistulas developed after a rupture of a thoracoabdominal aneurysm in a 73-year-old man and were successfully repaired in a one-stage procedure. This case demonstrates that operation can be successful even in this desperate situation. PMID- 9205190 TI - One-stage repair of truncus arteriosus, CAVC, and TAPVC. AB - An infant with truncus arteriosus, complete atrioventricular canal, and total anomalous pulmonary venous connection successfully underwent one-stage complete repair. Residual mitral valve regurgitation required reoperation after 12 days. The patient is doing well at 6 months' follow-up. Echocardiography demonstrates no residual defects, competent atrioventricular valves, and normal pulmonary pressure. This case illustrates the potential for successful one-stage repair even of associated complex heart defects involving venous, intracardiac, and arterial pathways. PMID- 9205191 TI - Thoracoscopic retrieval of foreign bodies after penetrating chest trauma. AB - Video-assisted thoracic surgery or thoracoscopy has proved to be valuable in many settings in thoracic surgery. The use of video-assisted thoracic surgery in trauma has been limited, especially with respect to penetrating trauma. We report the use of thoracoscopy to remove intrathoracic fragments of glass and avert the need for a thoracotomy. PMID- 9205192 TI - Intramural esophageal dissection. AB - A case of intramural esophageal dissection is reported and the literature reviewed. Patients with intramural esophageal dissection are usually women in their seventh or eighth decade. The most common presenting symptoms are chest pain, dysphagia, and hematemesis. The diagnosis is made by contrast esophagography, esophagoscopy, or both. Nonoperative therapy has proved to be uniformly successful. PMID- 9205193 TI - Tracheal stenosis treated with self-expanding nitinol stent. AB - A self-expanding nitinol stent was used in 2 patients with inoperable tracheal stenosis due to invasive malignant tumor of the trachea. One was a 70-year-old man with recurrent tumor from adenocarcinoma of the left lung, and the other was a 63-year-old man with recurrent tumor in mediastinal lymph nodes from esophageal cancer. The self-expanding nitinol stent is very useful and effective in inoperable tracheal stenosis due to intraluminal tumor invasion. PMID- 9205194 TI - Coronary artery bypass grafting after a bilateral lung volume reduction operation. AB - A 67-year-old man underwent coronary artery bypass grafting 31/2 months after a bilateral lung volume reduction operation for end-stage pulmonary emphysema. The principles of anesthetic management we have developed for use during volume reduction operations were applied with success in this individual and are described in detail. With the increasing application of this intervention as an alternative to lung transplantation, we anticipate further experience in the operative management of associated conditions after lung volume reduction operations. PMID- 9205195 TI - Traumatic arteriovenous fistula. AB - Traumatic arteriovenous fistula in the head and neck may present a difficult problem in management. We present a surgical case of traumatic arteriovenous fistula between the right subclavian artery and internal jugular vein with false aneurysm formation. Traumatic injury of the subclavian artery causing arteriovenous fistula with false aneurysm is a serious surgical emergency with appreciable morbidity and mortality that requires early recognition and prompt surgical intervention. PMID- 9205196 TI - Extracardiac right atrium-to-right ventricle homograft for uncorrectable tricuspid valve disease. AB - Surgically uncorrectable tricuspid valve disease in children is rare. However, when it happens the surgical options are very limited. Tricuspid valve replacement using a mechanical valve or stented bioprosthesis is impractical. Use of homografts in the "anatomic position" has its limitations. We report here the use of an extracardiac homograft connection between the right atrium and right ventricle in a 16-month-old boy in whom severe tricuspid valve stenosis developed after surgical repair of a complex ventricular septal defect associated with dextrocardia and anomalous systemic venous drainage. The patient remains well receiving no cardiac medication 12 months after the procedure. PMID- 9205197 TI - Facilitated exposure of the internal mammary artery in minimally invasive direct vision CABG. AB - A method is described to facilitate harvesting of the mammary artery in minimally invasive direct-vision coronary artery bypass grafting using a 10-cm anterior thoracotomy. Hoisting of the anterior thoracic wall with a modified retractor allows good exposure. Harvesting the mammary artery without the use of endoscopic tools was successful in all 10 cases. PMID- 9205198 TI - Correction of left superior vena cava draining to the left atrium using extracardiac techniques. AB - Intraatrial rerouting techniques have been the most common approaches to correcting left superior vena caval drainage to the left atrium in patients without atrial isomerism and with no connecting vein. Although these techniques have proved reliable and successful, there are cases in which extracardiac methods for managing this form of anomalous systemic drainage may be preferable. In the present report, we describe three extracardiac approaches to the correction of left superior vena cava draining to the left atrium. PMID- 9205199 TI - Alternative technique for the ostium primum defect repair: a free wall flap of right atrium. AB - An alternative surgical technique of repair of the ostium primum septal defect without the use of any patch is reported. The potential technical difficulties and surgical consideration are discussed. PMID- 9205200 TI - Linear segmental annuloplasty for mitral valve repair. AB - A method of posterior mitral annulus remodeling is presented. The posterior annulus is divided into three segments, each segment encircled by a suture that is passed in a tourniquet. Coaptation of the leaflets can be achieved by tightening the tourniquets while the ventricle is being filled. This technique is simple and quick, avoids the use of foreign material, and requires less expertise and judgment than traditional annuloplasties. PMID- 9205201 TI - Single-stage bilateral, video-assisted thoracoscopic lung volume reduction operation. AB - Lung volume reduction (LVR) produces significant clinical and objective improvement in selected patients with diffuse emphysema. Unilateral and bilateral approaches have been successfully employed. A median sternotomy approach is the standard for bilateral LVR, whereas video-assisted thoracoscopy has been used to perform unilateral LVR. Encouraging video-assisted thoracoscopic results with sequential, staged, bilateral LVR have been shown. This report describes an alternate technique of single-stage, bilateral LVR for end-stage emphysema. PMID- 9205202 TI - Pancoast (superior sulcus) tumors. AB - Primary carcinomas arising in the apex of the lung (Pancoast tumors) have attracted attention because of the characteristic syndrome that is produced by local extension into the chest wall and the brachial plexus. This article reviews the history of the treatment of this disease, the natural history of untreated patients, and the diagnosis of Pancoast tumors. The published data on results, prognostic factors, and technical aspects of treatment with combined irradiation and operation are examined, as well as those pertaining to treatment with irradiation alone. PMID- 9205203 TI - As originally published in 1989: Mitral annular calcification: a new technique for valve replacement. Updated in 1997. PMID- 9205204 TI - "Resource-based" practice expense: how we got where we are today. PMID- 9205205 TI - Pneumonectomy for inflammatory lung disease. PMID- 9205206 TI - Spontaneous echo contrast in mechanical valve prosthesis. PMID- 9205207 TI - Long-term follow-up after repair of coarctation of the aorta in adults. PMID- 9205208 TI - Platelet-rich plasma harvest in redo CABG. PMID- 9205209 TI - Psychological implications in the surgical treatment of pneumothorax. PMID- 9205210 TI - Minimally invasive direct atrial septal defect closure. PMID- 9205211 TI - Delayed tamponade after MIDCABG. PMID- 9205212 TI - The next medical revolution should be quality. PMID- 9205213 TI - Cystic fibrosis: 1997. PMID- 9205214 TI - Pulmonary arteries must be seen before they can be assessed. PMID- 9205215 TI - Chronic acalculous cholecystitis: are we diagnosing a disease or a myth? PMID- 9205216 TI - Laminar structures on MR images of articular cartilage. PMID- 9205217 TI - Accuracy of detection and measurement of renal calculi: in vitro comparison of three-dimensional spiral CT, radiography, and nephrotomography. AB - PURPOSE: To compare accuracy of three-dimensional (3D) spiral computed tomography (CT) performed without administration of contrast material with that of radiography and linear nephrotomography in detection and measurement of renal calculi. MATERIALS AND METHODS: Fifty renal calculi within an abdominal phantom were imaged with 3D spiral CT, radiography, and linear nephrotomography. Spiral CT data were analyzed with workstation-based 3D imaging software, with a thresholding procedure based on the maximally attenuating voxel within each calculus during measurement. Measurement accuracy and detection rates were compared according to modality. Conventional and magnification-corrected measurements from radiography and linear nephrotomography were included. RESULTS: Spiral CT depicted calculi and allowed determination of the collective two dimensional and 3D linear measurements statistically significantly more accurately than the other techniques; the mean linear measurement errors along individual axes did not exceed 3.6%. With 3D spiral CT, calculus volumes were determined with a mean error of -4.8%. CONCLUSION: 3D spiral CT enabled highly accurate determination of the volumes and all three linear dimensions of renal calculi. In addition, 3D spiral CT depicted calculi more sensitively than traditional techniques and provided new information and improved accuracy in the evaluation of nephrolithiasis. PMID- 9205218 TI - Ureteral calculi in patients with flank pain: correlation of plain radiography with unenhanced helical CT. AB - PURPOSE: To determine the sensitivity and specificity of plain radiography for the detection of ureteral calculi with use of unenhanced helical computed tomography (CT) as the standard of reference. MATERIALS AND METHODS: Plain radiographs and helical CT scans of 178 patients with acute flank pain were reviewed retrospectively. Three interpretations of plain radiographs were used: (a) Original reading. This was the report made at the time of the patient's evaluation before the patient underwent CT. (b) Blinded retrospective reading. Each plain radiograph was interpreted without knowledge of the CT findings. (c) Unblinded retrospective reading. The plain radiograph of each patient whose CT scan showed a stone was reviewed with the CT scan. RESULTS: The original reading had a sensitivity of 45% and a specificity of 77% for the detection of ureteral calculi. The blinded retrospective reading had a sensitivity of 59% and a specificity of 71%. The unblinded retrospective reading had a sensitivity of 59% (95% confidence interval: 47%, 70%). CONCLUSION: Plain radiography is of limited value for aiding the diagnosis of ureteral stones. All patients with acute flank pain for whom radiologic imaging is recommended can directly undergo unenhanced helical CT; plain radiographs need not be obtained first. PMID- 9205219 TI - Spiral noncontrast CT versus combined plain radiography and renal US after extracorporeal shock wave lithotripsy: cost-identification analysis. AB - PURPOSE: To investigate the costs of spiral computed tomography (CT) versus those of combined plain radiography and renal ultrasound (US) in screening for postprocedural complications after extracorporeal shock wave lithotripsy (ESWL). MATERIALS AND METHODS: Twenty-five adult patients who had undergone ESWL were prospectively examined with spiral CT, renal US, and plain abdominal radiography. Each examination was timed, and direct technical costs were calculated by using a procedural-based cost-accounting system. The combined cost of US and plain radiography was compared with the cost of spiral CT. RESULTS: The average time for spiral CT was 15.3 minutes compared with 37.2 minutes for combined US and plain radiography. The direct technical cost of spiral CT was $36.86 compared with $57.60 for combined US and plain radiography. Average examination times were varied to assess the effect on overall costs. Within reasonable time ranges, combined US and plain radiography cannot be cost equivalent to spiral CT. CONCLUSION: Spiral CT is faster and is associated with less direct technical cost than combined US and plain radiography when used to examine patients after ESWL, given the dependence of this model on time of examination. Further studies are needed to assess the relative accuracy of these alternative approaches. PMID- 9205220 TI - Adenoma malignum: MR imaging and pathologic study. AB - PURPOSE: To evaluate the clinical, pathologic, and magnetic resonance (MR) imaging findings in adenoma malignum, a rare variant of uterine cervical adenocarcinoma. MATERIALS AND METHODS: Medical records of all patients (n = 7) with adenoma malignum of the uterine cervix diagnosed pathologically between 1988 and 1996 were retrospectively reviewed. Unenhanced T1-weighted and T2-weighted images and gadolinium-enhanced T1-weighted MR images were evaluated, and findings were correlated with gross pathologic and microscopic features. RESULTS: In five of seven patients, enlargement of the cervix was seen. All lesions were detected as multiple cystic lesions that extended from the endocervical gland to the deep stroma of the cervix. They appeared isointense (n = 5) or slightly hyperintense (n = 2) relative to the uterus on T1-weighted images and markedly hyperintense relative to the uterus on T2-weighted images. Solid portions of variable size were seen between cystic lesions, and both the multiple cystic component and the solid portion were most apparent on the gadolinium-enhanced T1-weighted images. Microscopic parametrial invasion was seen in two patients but was not detected at MR imaging. CONCLUSION: Adenoma malignum was depicted on MR images as a multicystic mass with solid portions located in the deep cervical stroma. Gadolinium enhancement helped identify the solid portion of the tumor. PMID- 9205221 TI - Solid extratesticular masses evaluated with sonography: pathologic correlation. AB - PURPOSE: To determine if the sonographic appearance of solid extratesticular masses enables distinction of benign from malignant disease. MATERIALS AND METHODS: Sonograms of 19 patients with palpable testicular masses who underwent biopsy were reviewed retrospectively. Appearances of masses on sonograms were correlated with pathologic diagnoses. RESULTS: All masses were well defined and ranged in size from 5.7 to 66.7 mm (mean, 21 mm). On the sonograms, five masses were within the epididymis, and six were distinct from it; seven cases were indeterminate. The epididymis was surgically absent in the remaining patient. Sonographic echogenicity ranged from hypoechoic to hyperechoic relative to that of the testis. At pathologic evaluation, there were 16 (84%) benign and three (16%) malignant lesions. Benign lesions consisted of six adenomatoid tumors, two lipomas, two epidermoid inclusion cysts, two cases of sarcoidosis, and one case each of sperm granuloma, spermatic cord leiomyoma, benign inflammatory nodule, and fibroma. The malignant lesions consisted of scrotal wall liposarcoma, epididymal leiomyosarcoma, and recurrent spindle cell malignancy of the spermatic cord. No sonographic features of masses were useful for distinguishing benign from malignant lesions. CONCLUSION: The frequency of malignancy (16%) contrasts with prior reports that suggest a very low rate of malignancy among these masses. Sonography is useful for identifying the extratesticular location of a mass but not for distinguishing the nature of the lesion. PMID- 9205222 TI - Multicenter trial validation of a camera-based method to measure Tc-99m mercaptoacetyltriglycine, or Tc-99m MAG3, clearance. AB - PURPOSE: To evaluate an improved camera-based method for calculating the clearance of technetium-99m mercaptoacetyltriglycine (MAG3) in a multicenter trial. MATERIALS AND METHODS: Tc-99m MAG3 scintigraphy was performed in 49 patients at three sites in the United States and Canada. The percentage of the injected dose of Tc-99m MAG3 in the kidney at 1-2, 1.0-2.5, and 2-3 minutes after injection was correlated with the plasma-based Tc-99m MAG3 clearances. The data were combined with the results obtained in 20 additional patients in a previously published pilot study. RESULTS: Regression models correlating the plasma-based Tc 99m MAG3 clearance with the percentage uptake in the kidney for each time interval were developed; there was no statistically significant difference among sites in the regression equations. Correction for body surface area statistically significantly (P < .005) improved the correlation coefficient for each time interval. For the 1.0-2.5-minute interval, the body surface area-corrected correlation coefficient for the four combined sites was .87, and it improved to .93 when one outlier was omitted from the analysis. Similar results were obtained with the other time intervals. Independent processing by two observers showed no clinically important differences in the percentage dose in the kidney or in relative function. CONCLUSION: An improved camera-based method to calculate the clearance of Tc-99m MAG3 was validated in a multicenter trial. PMID- 9205223 TI - Differentiation between recurrent tumor and benign conditions after treatment of gynecologic pelvic carcinoma: value of dynamic contrast-enhanced subtraction MR imaging. AB - PURPOSE: To compare dynamic contrast material-enhanced subtraction and T2 weighted spin-echo (SE) magnetic resonance (MR) imaging in the differentiation of fibrosis from tumor recurrence during the follow-up of treated gynecologic pelvic malignancy. MATERIALS AND METHODS: Thirty-four patients (aged 24-82 years) with 18 benign and 35 malignant lesions confirmed by means of surgery (n = 18), biopsy (n = 25), or 18-month follow-up examination (n = 10) underwent dynamic contrast enhanced subtraction and T2-weighted SE MR imaging. Contrast material enhancement of an abnormal pelvic structure within the first 90 seconds on dynamic contrast enhanced subtraction images or high signal intensity on T2-weighted SE images was considered indicative of malignancy. RESULTS: The sensitivity, specificity, accuracy, and positive and negative predictive values were 91%, 67%, 83%, 86%, and 86%, respectively, for dynamic contrast-enhanced subtraction imaging and 91%, 22%, 68%, 70%, and 57%, respectively, for T2-weighted SE imaging. More lesions were correctly classified with dynamic contrast-enhanced subtraction imaging than with T2-weighted SE imaging (P < .01). CONCLUSION: Dynamic contrast-enhanced subtraction imaging is more accurate than T2-weighted SE imaging for differentiating fibrosis from tumor recurrence during the follow-up of treated gynecologic pelvic malignancy. However, use of both sequences is recommended. PMID- 9205224 TI - Renal angiomyolipoma: selective arterial embolization--effectiveness and changes in angiomyogenic components in long-term follow-up. AB - PURPOSE: To evaluate the efficacy of selective arterial embolization in symptomatic renal angiomyolipoma (AML) and the change in angiomyogenic components during long-term follow-up after embolization. MATERIALS AND METHODS: Fourteen adult patients with symptomatic AMLs underwent 16 selective arterial embolizations. The embolic materials used were absolute alcohol with (n = 5) or without (n = 3) iodized oil, Gianturco coils (n = 4), and polyvinyl alcohol foam powder with gelatin sponge (n = 2). Follow-up ultrasonography and computed tomography (CT) were performed in six and 14 patients, respectively. The effectiveness of selective arterial embolization was evaluated on the basis of the area of the angiomyogenic components in the AML on initial and follow-up images and clinical improvement. RESULTS: All patients showed devascularization of the tumor on the postembolization angiograms. In 13 patients, clinical symptoms disappeared. The follow-up period was 7-72 months (mean, 33 months). One patient underwent nephrectomy at 7 months after embolization because of a large cystic lesion found at 1 month. In long-term CT follow-up (> or =12 months) in 12 patients, nearly all angiomyogenic components disappeared, but fatty components partially shrank with liquefactive necrosis in tumors. CONCLUSION: Selective arterial embolization is an effective and safe treatment of AML. The angiomyomatous components crucial for the prevention of bleeding were very sensitive to the embolization. PMID- 9205225 TI - Chronic mesenteric ischemia: use of in vivo MR imaging measurements of blood oxygen saturation in the superior mesenteric vein for diagnosis. AB - PURPOSE: To determine if dogs and humans with chronic mesenteric ischemia demonstrate a decrease in the percentage of oxygenated hemoglobin (%HbO2) in the superior mesenteric vein (SMV) after a meal. MATERIALS AND METHODS: In 10 dogs, ameroid rings were surgically implanted around the superior mesenteric arteries to create gradual stenosis. Pre- and postoperative angiograms and pre- and postprandial magnetic resonance (MR) oximetry measurements of the SMV %HbO2, with flow-independent T2 measurements of venous blood, were obtained at different times. In 10 patients with atherosclerotic disease and six patients with symptomatic chronic mesenteric ischemia, the same measurements were obtained after at least 6 hours of fasting and at 15, 35, and 45 minutes after ingestion of a liquid nutritional supplement. RESULTS: In seven dogs, the postprandial SMV %HbO2 increased an average of 2.5% +/- 0.8 before surgery and decreased an average of 6.3% +/- 2.1 when hemodynamically significant (>70%) stenosis of the superior mesenteric artery developed 7-14 days after surgery. In the 10 patients without ischemia, the SMV %HbO2 increased by 4.6% +/- 0.6, whereas in the symptomatic patients a postprandial decrease of 8.8% +/- 0.7 occurred (P < .0001). CONCLUSION: Measurement of the SMV %HbO2 with MR oximetry is a promising test for diagnosis of chronic mesenteric ischemia. PMID- 9205226 TI - Treatment of abdominal aortic aneurysms with transfemoral placement of stent grafts: complications and secondary radiologic intervention. AB - PURPOSE: To determine the rate of complications of transluminally placed endovascular stent-grafts in patients with abdominal aortic aneurysms and to assess efficacy of secondary radiologic intervention. MATERIALS AND METHODS: In 28 patients, covered nitinol stents were implanted. Frequency and outcome of complications were evaluated after stent-graft placement (mean follow-up, 8.2 months). RESULTS: In 27 patients, 11 tube and 16 bifurcated grafts were implanted successfully. In one (4%) patient, distal migration of a bifurcated graft necessitated conversion to standard open repair. Technical success rates (successful deployment of the device and complete exclusion of the aneurysm at intraoperative angiography) for bifurcated and tube grafts were 24% (four of 17 patients) and 91% (10 of 11 patients). After surgery, leaks were seen in six bifurcated and five tube grafts. After secondary intervention, the final exclusion rate for bifurcated and tube grafts was 88% (15 of 17 patients) and 100% (11 of 11 patients), respectively; the overall success rate was 93% (26 of 28 patients) for exclusion of aortic abdominal aneurysms. Stent-graft thrombosis necessitated local thrombolysis in one patient. Overlapping of accessory renal arteries occurred in two patients. CONCLUSION: Complications after stent-graft placement in abdominal aortic aneurysms are frequent. Secondary radiologic intervention is successful in most cases. PMID- 9205227 TI - Meta-analysis of the results of percutaneous transluminal angioplasty and stent placement for aortoiliac occlusive disease. AB - PURPOSE: To estimate and compare the results of percutaneous transluminal angioplasty (PTA) and stent placement to treat aortoiliac occlusive disease. MATERIALS AND METHODS: A meta-analysis was performed of data in six PTA studies (1,300 patients) and eight stent placement studies (816 patients) published in 1990 or later that met the inclusion criteria. Proportions were combined by means of a random-effects model. Failure-time data were pooled with and pooled without adjustment for differences in case mix. RESULTS: The immediate technical success rate in the PTA group was 91%; the rate was higher in the stent group (96%), but the difference was not statistically significant [corrected]. Complication and mortality rates were not statistically significantly different. Analyzed data included technical failures and were adjusted for lesion type and disease severity. Four-year primary patency rates were 65% for stenoses versus 54% for occlusions after PTA to treat claudication and were 53% for stenoses versus 44% for occlusions after PTA to treat critical ischemia. These rates were 77% for stenoses versus 61% for occlusions after stent placement to treat claudication and 67% for stenoses versus 53% for occlusions after stent placement to treat critical ischemia. The risk of long-term failure was reduced by 39% after stent placement compared with PTA. CONCLUSION: Stent placement and PTA yielded similar complication rates, but the technical success rate was higher after stent placement and the risk of long-term failure was reduced. PMID- 9205228 TI - Patients with thrombocytopenia: outcome of radiologic placement of central venous access devices. AB - PURPOSE: To determine prospectively the outcome of radiologic placement of central venous access devices in patients with thrombocytopenia. MATERIALS AND METHODS: In 105 patients, 87 catheters, 10 arm port systems, and eight chest port systems were placed radiologically. Devices and patients were separated into group A (n = 37; platelet count < 50,000 x 10(6)/L [50 x 10(9)/L]), group B (n = 35; platelet count, 50-100,000 x 10(6)/L [0.05-100 x 10(9)/L]), and group C (n = 33; platelet count, > 100,000 x 10(6)/L [100 x 10(9)/L]). Patients in group A received platelet transfusions during implantation. Patients were followed up for up to 8 weeks (mean, 41.2 days). Success and complication rates (immediate and delayed) were determined for each group. RESULTS: There were no bleeding complications that necessitated intervention in patients with thrombocytopenia (groups A and B). There was no statistically significant difference in complication rates per "catheter days" among the three groups (4.2 per 1,000 catheter days in group A, 4.6 per 1,000 catheter days in group B, and 5.2 per 1,000 catheter days in group C). Postprocedure platelet counts increased only slightly (mean, 11,500 x 10(6)/L [11.5 x 10(9)/L]) in patients in group A. CONCLUSION: Radiologic placement of central venous access devices can be performed safely in patients with thrombocytopenia. PMID- 9205229 TI - Liver biopsy: effects of biopsy needle caliber on bleeding and tissue recovery. AB - PURPOSE: To evaluate different-caliber biopsy cutting needles in terms of the benefits and potential risk of bleeding in a swine model. MATERIALS AND METHODS: A total of 190 sequential liver biopsy specimens were obtained in 11 Yorkshire pigs (weight, 50-70 lb [22.5-31.5 kg]) by using 14-, 18-, and 20-gauge cutting needles. For each biopsy procedure, blood loss was determined by weighing sponges used to absorb bleeding, and sample-tissue DNA content was measured with spectrofluorometry. Analysis of variance was used to compare results. RESULTS: The larger the caliber of needle, the greater the absolute blood loss (for 14 gauge, 1.69 g; for 18-gauge, 0.74 g; for 20-gauge, 0.32 g) and DNA content per sample (for 14 gauge, 40.38 microg; for 18-gauge, 12.18 microg; for 20-gauge, 5.86 microg). The ratio of blood loss to amount of DNA recovered did not differ among the different-caliber needles. To obtain the same amount of diagnostic tissue, more passes were needed with the smaller-caliber needles. CONCLUSION: Use of larger-caliber needles is more efficient despite the greater amount of blood loss, because more tissue can be recovered and because fewer passes are necessary, which reduces the chances of complications. PMID- 9205230 TI - Acalculous gallbladder disease: US evaluation after slow-infusion cholecystokinin stimulation in symptomatic and asymptomatic adults. AB - PURPOSE: To evaluate the measurement with ultrasonography (US) of the gallbladder ejection fraction after slow-infusion cholecystokinin stimulation in patients with biliary symptoms and in individuals without symptoms. MATERIALS AND METHODS: Gallbladder volumes were calculated in 60 healthy volunteers after a 30-minute infusion of sincalide. The time to maximum response, the gallbladder ejection fraction, and the rate of initial contraction were obtained at US. A total of 100 symptomatic patients were evaluated with this technique. Reference standards included surgical outcome or results of clinical follow-up of at least 1 year. RESULTS: The average ejection fraction +/- 2 standard deviations was 80% +/- 30. A fraction greater than 60% was considered to be a normal response to cholecystokinin stimulation. There was no statistically significant sex difference. Slow infusion did not produce any side effects. A sensitivity of 75% and a specificity of 100% for determination of gallbladder ejection fraction at US were obtained in patients with surgical and histopathologic proof of disease. CONCLUSION: The slow-infusion method is reliable, safe, and reproducible in evaluating gallbladder contraction. The cholecystokinin-stimulated gallbladder ejection fraction test may be useful in determining which patients could benefit from surgery. PMID- 9205232 TI - US-guided left-sided biliary drainage: nine-year experience. AB - PURPOSE: The feasibility and safety of left-sided biliary drainage with ultrasound (US) guidance were studied prospectively. MATERIALS AND METHODS: From July 1987 to July 1996, 208 consecutive patients underwent US-guided biliary drainage; all were evaluated for left-sided drainage. Drainage procedure was begun with puncture of the hepatic duct branch of the lateral segment of the left lobe when the branch was well visualized with US; otherwise, a right-sided approach was used. When the hepatic duct branch diameter was greater than 3 mm, puncture was performed with an 18-gauge needle; smaller branches were punctured with 21-gauge needles. RESULTS: In 147 (71%) patients, the left hepatic duct branch was well visualized with US, and the branch diameter was greater than 3 mm. In these patients, left-sided drainage with use of an 18-gauge needle was successful. In 26 (12%) patients, the left hepatic duct branch diameter was less than 3 mm, and drainage was initiated with a 21-gauge needle. In six (23%) of these 26 patients, left-sided drainage was unsuccessful, but five of these patients underwent successful US-guided drainage from the right hepatic duct branch. Two patients died of septic shock within 72 hours of completed drainage. Three patients experienced severe hemobilia. CONCLUSION: US-guided left-sided biliary drainage is a highly successful and safe method when the left hepatic duct branch diameter is greater than 3 mm. PMID- 9205231 TI - Analysis of biliary drainage in the caudate lobe of the liver: comparison of three-dimensional CT cholangiography and rotating cine cholangiography. AB - PURPOSE: To evaluate the bile duct anatomy of the caudate lobe without disease involvement with use of three-dimensional (3D) cholangiography and to compare the usefulness of this technique with that of rotating cine cholangiography. MATERIALS AND METHODS: In 12 patients with obstructive jaundice but without lesions at the hepatic hilum who underwent percutaneous transhepatic biliary drainage, serial examination was performed with cine cholangiography and helical computed tomography (CT). From helical CT scans, 3D cholangiograms were reconstructed. Cine and 3D cholangiograms were evaluated and compared simultaneously. RESULTS: In the 12 patients, 40 branches of the caudate lobe were detected with 3D cholangiography (mean, 3.3 branches per patient), while 31 were detected with cine cholangiography (mean, 2.6 branches per patient). The difference in detection rate was significant (P < .01). Nine (23%) of 40 branches were detected with 3D cholangiography alone, and all 31 branches detected with cine cholangiography were also detected with 3D cholangiography. CONCLUSION: 3D cholangiography was superior to cine cholangiography in assessment of bile duct anatomy of the caudate lobe of the liver because 3D cholangiography eliminated the overlap of different branches of the bile duct. PMID- 9205233 TI - Fibroadenomas: MR imaging appearances with radiologic-histopathologic correlation. AB - PURPOSE: To identify histopathologic correlates for the varied magnetic resonance (MR) imaging appearances of fibroadenomas. MATERIALS AND METHODS: Twenty-three fibroadenomas in 21 patients (aged 23-66 years) examined with gadolinium-enhanced MR imaging were graded for signal intensity on T2-weighted images, contrast material enhancement, shape, and internal septations and were correlated with histopathologic findings. RESULTS: Fibroadenomas demonstrated high T2 signal intensity with enhancement (n = 11), low T2 signal intensity with enhancement (n = 3), or low T2 signal intensity without enhancement (n = 9). Low T2 signal intensity and lack of enhancement were associated with more sclerotic stroma and older patient age. Lesion shape was lobular, oval, or round in 19 of 23 fibroadenomas (83%). Internal septations were identified within nine of 14 enhancing fibroadenomas (64%) and appeared to correlate with collagenous bands at histopathologic analysis. CONCLUSION: Fibroadenomas demonstrate marked histopathologic variability. The resultant variability in the MR appearance limits the ability to distinguish between benign and malignant masses on the basis of signal intensity and enhancement alone. Lobulation and internal septation, which appear to reflect intrinsic growth patterns of fibroadenomas, may provide a more reliable basis for distinction. PMID- 9205234 TI - Characteristics of breast carcinomas missed by screening radiologists. AB - PURPOSE: To determine whether breast cancers missed at screening mammography have distinguishing characteristics from those of detected cancers. MATERIALS AND METHODS: The mammograms of 146 women with mammographically identifiable breast cancer were viewed independently by two radiologists who were blinded as to whether the cancer had been missed or detected (group 1 lesions, missed cancers; group 2 lesions, detected cancers) at screening. The mammographic lesions were characterized as to location, size, density, type, and visibility on two views. RESULTS: A significant difference between missed and detected cancers was found for diameter (P = .03), number of views (P < .0017), and density (P = .0007). Stepwise multivariable logistic regression showed that density (P = .01) and the number of views (P = .03) but not diameter (P = .27) were independently significant in distinguishing the groups. No statistically significant difference was found between the two groups for lesion type (P = .32 for reader 1 and P = .27 for reader 2) or location (P = .86 for reader 1 and P > .96 for reader 2). CONCLUSION: Missed cancers were statistically significantly lower in density and more often seen on only one of two views than detected cancers. PMID- 9205235 TI - Dilated duct pattern at mammography. AB - PURPOSE: To determine the importance of a dilated duct pattern at mammography. MATERIALS AND METHODS: Mammograms obtained in 46 women with histopathologically proved, asymmetrically dilated ducts were retrospectively studied. The laterality and location of the asymmetrically dilated duct, the presence of branching, and associated findings such as microcalcifications, nipple discharge, and interval change were evaluated. RESULTS: Eleven patients (24%) had malignant results (ductal carcinoma in situ or invasive ductal carcinoma). Among these, six (54%) had suspicious microcalcifications. Nonsubareolar location and interval change are significant (P = .04) variables associated with malignancy. CONCLUSION: Mammographic asymmetrically dilated ducts in a nonsubareolar area that are associated with interval change, suspicious microcalcifications, or both warrant biopsy. PMID- 9205236 TI - Breast intraductal masses: US-guided fine-needle aspiration after galactography. AB - PURPOSE: To evaluate ultrasonographic (US)-guided fine-needle aspiration (FNA) of intraductal masses performed immediately after galactography and to compare cytologic findings from US-guided FNA with those from nipple discharge. MATERIALS AND METHODS: In 36 patients with nipple discharge from a single duct in one breast and intraductal masses diagnosed at galactography, US was performed to detect intraductal lesions and perform FNA before removal of the galactographic catheter. Cytologic analysis of nipple discharge, excisional biopsy, and histopathologic evaluation were performed in all patients. RESULTS: Cytologic analysis revealed 23 nonpapillary benignancies, seven papillomas, five indeterminate cases, and one carcinoma. US-guided FNA cytologic analysis revealed 16 papillomas, 10 nonpapillary benignancies, five indeterminate cases, and three carcinomas. The two carcinomas misdiagnosed as papillomas at US-guided FNA cytologic analysis were papillary in situ carcinomas, while the three carcinomas correctly identified were invasive (only one was detected with cytologic analysis of nipple discharge). With cytologic analysis of nipple discharge, nine (25%) of 36 diagnoses were correct, and with US-guided FNA, 18 (50%) were correct (P = .0352). CONCLUSION: Compared with cytologic analysis of nipple discharge, US guided FNA cytologic analysis seems to add useful information for tailored surgical planning. PMID- 9205237 TI - Impalpable breast cysts: utility of cytologic examination of fluid obtained with radiologically guided aspiration. AB - PURPOSE: To evaluate the utility of cytologic analysis of fluid obtained from impalpable breast cysts by means of radiologically guided aspiration. MATERIALS AND METHODS: The authors retrospectively reviewed the reports of cytologic examinations of fluid obtained with sonographically or mammographically guided aspiration of 660 impalpable breast cysts in 583 women during 3 1/2 years. RESULTS: No malignant cells (541 cysts) or insufficient cellular material (86 cysts) was seen with cytologic examination of 95% of the aspirates. Atypical cells were seen with cytologic examination of fluid from 33 (5%) lesions. None of these 33 lesions were found to represent malignancy at the time of surgical excision (n = 9) or during clinical follow-up (n = 24). CONCLUSION: Routine cytologic examination is unnecessary if the fluid obtained with radiologically guided aspiration from impalpable breast cysts is not bloody. PMID- 9205238 TI - Mammographic findings after 14-gauge automated needle and 14-gauge directional, vacuum-assisted stereotactic breast biopsies. AB - PURPOSE: To compare findings from first imaging follow-up mammography for breast lesions shown to be benign at stereotactic biopsy with 14-gauge automated needles or 14-gauge directional, vacuum-assisted probes. MATERIALS AND METHODS: In 495 stereotactic breast biopsies, the mammographic appearance of the biopsy site or target lesion was evaluated at first imaging follow-up with a four-point scale (1 = progression of lesion or suspicious interval change [repeat biopsy], 2 = no clinically relevant change, 3 = interval decrease in size of lesion or number of microcalcifications, and 4 = no residual mammographic lesion). An automated needle was used in 363 biopsies and a directional, vacuum-assisted probe was used in 132 biopsies. Patient and lesion variables and time to first imaging follow-up were compared for the two techniques. RESULTS: No biopsy site or target lesion was assigned a score of 1 at first imaging follow-up with either technique. No lesion was referred for repeat biopsy because of the mammographic appearance at first imaging follow-up. First imaging follow-up was performed an average of 6.6 months for the directional, vacuum-assisted biopsies and 8.6 months for the automated needle biopsies (P < .0001). This difference reflected a difference in scheduling methods. CONCLUSION: Directional, vacuum-assisted and automated needle breast biopsies produced no distortion or suspicious interval change at the biopsy site at the first follow-up mammographic examination. PMID- 9205239 TI - Peripheral pulmonary arteries: optimization of the spiral CT acquisition protocol. AB - PURPOSE: To analyze the influence of collimation on identification of segmental and subsegmental pulmonary arteries on spiral computed tomographic (CT) scans. MATERIAL AND METHODS: Contrast material-enhanced spiral CT scans were retrospectively analyzed. Patients in group A (n = 20) underwent CT with 3-mm collimation, 1.00 second per revolution, and pitch of 1.7; those in group B (n = 20) underwent CT with 2-mm collimation, 0.75 second per revolution, and pitch of 2.0. A total of 800 segmental (20 arteries per patient) and 1,600 subsegmental (40 arteries per patient) arteries were assessed. RESULTS: The mean number of analyzable segmental arteries per patient was greater in group B patients (18.6 of 20.0 [93%]) than that in group A patients (17.0 of 20.0 [85%]) (P < .001). The mean number of analyzable subsegmental arteries per patient was greater in group B patients (24.6 of 40.0 [61%]) than that in group A patients (14.8 of 40.0 [37%]) (P < .0001). Frequency of identification on CT scans of 13 of the 40 subsegmental arteries was improved in group B compared with group A patients (P < .0001). CONCLUSION: Spiral CT with 2-mm collimation at 0.75 second per revolution enables marked improvement in the analysis of segmental and subsegmental pulmonary arteries. PMID- 9205240 TI - Reversible cystic disease associated with pulmonary tuberculosis: radiologic findings. AB - PURPOSE: The authors describe radiologic findings of cystic lung disease associated with pulmonary tuberculosis. MATERIALS AND METHODS: Three immunocompetent women with pulmonary tuberculosis were seen with acute respiratory failure and diffuse bilateral pulmonary opacity depicted at admission chest radiography. The women did not have acquired immunodeficiency syndrome, or AIDS. They were aged 26, 27, and 52 years. RESULTS: Follow-up radiography and computed tomography showed multiple air-filled cystic lesions in both lungs, which disappeared almost completely after antituberculous chemotherapy (isoniazid, rifampin, ethambutol hydrochloride, and pyrazinamide). CONCLUSION: Pulmonary tuberculosis manifested with reversible multiple cystic lung disease in three immunocompetent patients. PMID- 9205241 TI - Mycobacterium kansasii pulmonary infection in patients with AIDS: spectrum of chest radiographic findings. AB - PURPOSE: To determine the chest radiographic findings and clinical manifestations of Mycobacterium kansasii pulmonary infection in patients with acquired immunodeficiency syndrome (AIDS). MATERIALS AND METHODS: Criteria for diagnosis included two or more positive cultures from respiratory sources, pulmonary symptoms or fever, and no other identifiable cause of pulmonary disease. Chest radiographs at initial examination and follow-up were evaluated for parenchymal opacities, cavitation, adenopathy, and pleural effusions. Medical records were reviewed for clinical signs and symptoms, CD4 cell count, presence of additional pathogens, and response to antimycobacterial therapy. RESULTS: Of 96 patients, 16 (17%) satisfied all criteria for M kansasii pulmonary infection. The mean CD4 cell count was 24/mm3. Twelve patients (75%) demonstrated alveolar opacities, only three (19%) of which were cavitary. Interstitial opacities (6%) and pleural effusions (12%) were uncommon. Four (25%) patients had thoracic lymphadenopathy, which was the only positive radiographic finding in two patients. Fourteen patients were treated for M kansasii, and 10 (71%) showed clinical and radiographic improvement. CONCLUSION: Patients with AIDS and pulmonary M kansasii frequently demonstrate focal alveolar opacities. Symptomatic patients with pulmonary nontuberculous mycobacteria should be presumptively treated for pulmonary M kansasii until final culture results are available. PMID- 9205242 TI - Shin splints: MR appearance in a preliminary study. AB - PURPOSE: To investigate the magnetic resonance (MR) imaging appearance of activity-related lower leg pain (shin splints syndrome) and evaluate the relative involvement of bone and soft tissues. MATERIALS AND METHODS: Nineteen patients with activity-related lower leg pain and tenderness on palpation along the posteromedial tibia (shin splints) underwent clinical examination and MR imaging. Five also underwent plain radiography. MR findings were compared with patient demographics, clinical findings, and plain radiographs when available. RESULTS: Four MR patterns were identified: normal appearance (n = 7), periosteal fluid only (n = 5), abnormal marrow signal intensity (n = 5), and stress fracture (n = 2). Increased symptom duration correlated strongly with a normal MR image (P = .002). Plain radiographs appeared normal in all five patients for whom they were available. CONCLUSION: Patients with acute shin splints have a spectrum of MR findings, which suggests this clinical entity is part of a continuum of stress response in bone. The strong association between chronic symptoms and a normal appearing MR image implies that this modality has less utility in these patients. PMID- 9205243 TI - Anterior horn of the lateral meniscus: another potential pitfall in MR imaging of the knee. AB - PURPOSE: To demonstrate that speckled increased signal intensity at the anterior horn of the lateral meniscus near its central attachment site on sagittal magnetic resonance (MR) images of the knee is a normal finding. MATERIALS AND METHODS: In 22 patients (17 male and five female patients; age range, 13-74 years; mean, 38 years) who underwent arthroscopy after MR imaging, knee MR images that showed speckled increased signal intensity at the anterior horn of the lateral meniscus near its central attachment site on two consecutive sagittal proton-density-weighted images were selected for retrospective review. In addition, a review of 11 knee MR examinations of nine healthy volunteers (five men and four women; age range, 27-43 years; mean, 34 years) was performed. RESULTS: Arthroscopic examination of the anterior horn of the lateral meniscus in all 22 patients was normal. Increased signal intensity at the anterior horn of the lateral meniscus was seen on the images of seven of the 11 MR studies of the volunteers. CONCLUSION: Increased signal intensity at the anterior horn of the lateral meniscus near its central attachment site on knee MR images does not represent a meniscal tear. PMID- 9205244 TI - Posterolateral rotatory instability of the elbow: usefulness of MR imaging in diagnosis. AB - PURPOSE: To evaluate the efficacy of magnetic resonance (MR) imaging in the assessment of the normal and abnormal ulnar band of the lateral collateral ligament for diagnosis of posterolateral rotatory instability. MATERIALS AND METHODS: In nine symptomatic patients and nine asymptomatic subjects, MR imaging was performed with three-dimensional gradient-recalled and fast spin-echo sequences. The nine patients had clinical symptoms suggestive of subtle elbow instability. RESULTS: The components of the lateral collateral ligament were identified; tears of the ulnar band were noted in all symptomatic patients. The anterior fibers of the lateral collateral ligament, including the annular ligament, were intact. All symptomatic patients subsequently underwent surgical exploration and reconstruction. Positive clinical findings were demonstrated at examination performed while the patients were under anesthesia. All tears of the ulnar band were confirmed. CONCLUSION: With use of appropriate pulse sequences, MR imaging is an effective tool in the preoperative, noninvasive diagnosis of posterolateral rotatory instability. PMID- 9205245 TI - MR imaging of the rotator cuff tendon: interobserver agreement and analysis of interpretive errors. AB - PURPOSE: To evaluate accuracy in interpretation of shoulder magnetic resonance (MR) images and interobserver agreement and to characterize the types of errors. MATERIALS AND METHODS: Five radiologists with varying experience independently and retrospectively twice interpreted the MR images of 222 symptomatic patients who underwent both MR imaging and shoulder arthroscopy. The first interpretation was a blind review; the second was with knowledge of the arthroscopic findings. RESULTS: For full-thickness tears, the sensitivity, specificity, and accuracy were 84%-96%, 94%-98%, and 92%-97%, respectively, and for partial tears, 35%-44%, 85%-97%, and 77%-87%, respectively. There was no statistically significant difference between readers in the detection of partial or full-thickness tears. There was a statistically significant difference between readers in the no-tear category. Cohen kappa values generally indicated improved interobserver agreement proportional to the readers' experience (full-thickness tears, 0.731-0.881; partial tears, 0.168-0.443). CONCLUSION: Diagnosis of a full-thickness tear can be learned to a high degree of accuracy. Despite the radiologist's level of experience and knowledge of the arthroscopic findings, the sensitivity for diagnosis of partial tears is poor. PMID- 9205246 TI - Vertebral compression fractures in multiple myeloma. Part I. Distribution and appearance at MR imaging. AB - PURPOSE: To study the appearance and distribution of vertebral compression fractures on magnetic resonance (MR) images in patients with multiple myeloma. MATERIALS AND METHODS: Two hundred twenty-four vertebral compression fractures were studied on 216 sagittal T1-weighted spin-echo and T2*-weighted gradient-echo MR images of the thoracolumbar spine obtained before and during treatment in 37 patients with multiple myeloma. Vertebral compression fractures observed at diagnosis and during follow-up were determined as being benign- or malignant appearing at MR imaging according to literature criteria, and their distribution along the spine was recorded. RESULTS: One hundred forty-nine (67%) of the 224 vertebral compression fractures appeared benign; 75 (33%) appeared malignant. Of the 37 patients, 14 (38%) had only benign-appearing vertebral compression fractures at diagnosis. One hundred five fractures (87%) were observed between T 6 and L-4, and 112 (50%) occurred between T-11 and L-3. Eight (4%) vertebral compression fractures involved the upper three thoracic vertebrae. CONCLUSION: Most vertebral compression fractures in patients with multiple myeloma appear benign at MR imaging, and their distribution is similar to that observed in osteoporotic fractures. The possibility of multiple myeloma should not be excluded in patients with benign-appearing vertebral compression fractures at MR imaging. PMID- 9205247 TI - Vertebral compression fractures in multiple myeloma. Part II. Assessment of fracture risk with MR imaging of spinal bone marrow. AB - PURPOSE: To determine the utility of bone marrow magnetic resonance (MR) imaging in the assessment of risk of vertebral compression fractures in patients with multiple myeloma. MATERIALS AND METHODS: In 50 patients with stage III multiple myeloma, 280 MR examinations of the thoracolumbar spine obtained at diagnosis and during treatment (mean follow-up, 28 months) were analyzed to determine MR patterns of bone marrow involvement before treatment and the occurrence of vertebral compression fracture at follow-up. Four MR patterns of marrow involvement were determined: A, normal marrow appearance; B, fewer than 10 focal lesions; C, more than 10 focal lesions; and D, diffuse infiltration. Fracture free survival was compared according to these patterns. RESULTS: During follow up, 131 vertebral compression fractures appeared in 37 patients. Patients with pattern A (n = 10) or B (n = 16) had significantly longer fracture-free survival before occurrence of the first, second, and third fractures than those with pattern C or D (P < 10(-5)). Relative risks of first, second, and third fracture occurrence for patients with pattern C or D compared with those with pattern A or B were 6.2, 9.1, and 11.0, respectively. CONCLUSION: Determination of MR patterns of spinal bone marrow involvement is a potential relevant factor to predict the risk of vertebral fractures in patients with stage III multiple myeloma. PMID- 9205248 TI - Locally advanced head and neck cancer: combined chemotherapy and radical radiation therapy for organ and function preservation (interim report). AB - PURPOSE: This phase II study was performed to assess the feasibility of organ preservation after combined chemotherapy and radical radiation therapy in patients with resectable, locally advanced head and neck cancer. MATERIALS AND METHODS: Twenty-four patients had surgically resectable stage III (n = 9) and stage IV (n = 15) squamous cell carcinoma of the head and neck. Initially, they received two to three courses of neoadjuvant chemotherapy (5-fluorouracil and cisplatin). Patients in whom response was seen were then treated definitively with two courses of cisplatin chemotherapy administered concomitantly with radical radiation therapy. Patients in whom no response was seen underwent salvage surgery or other standard therapy. RESULTS: The response rate after induction chemotherapy was 84% (including 42% complete remission). Complete remission after concomitant chemotherapy and radical radiation therapy was 83%. After a median follow-up of 18 months (maximum, 3.5 years), 18 patients (75%) remained recurrence free. Grade 3 or 4 treatment-related toxicity was experienced, but there was no treatment-related mortality. CONCLUSION: Combined chemotherapy and radical radiation therapy used in this study resulted in organ and function preservation in the majority of patients with resectable stage III and IV head and neck cancer. PMID- 9205249 TI - Differences in dosimetry and toxicity between definitive and postprostatectomy radiation therapy. AB - PURPOSE: To compare the volumes of normal tissue irradiation and toxicity in patients treated with definitive radiation therapy or postprostatectomy radiation therapy. MATERIALS AND METHODS: Results were analyzed prospectively in 49 patients who underwent definitive or postprostatectomy radiation therapy after three-dimensional conformal treatment planning. Dose-volume histograms and the effective area irradiated were analyzed. Patient charts were reviewed and scored for acute and chronic toxicity. The mean bladder volume was 200 mL in the definitive group and 226 mL in the postprostatectomy group. The mean rectal volume was 87 mL in the definitive group and 81 mL in the postprostatectomy group. RESULTS: The dose to 25% of the rectal volume was 91% and 86% in the definitive and postprostatectomy groups, respectively. The dose to 25% of the bladder volume was 89% in the definitive group and 82% in the postprostatectomy group. The incidence of acute and chronic gastrointestinal and chronic genitourinary toxicity was similar in both groups, but acute genitourinary toxicity in the postprostatectomy group was significantly lower (P = .026). The volume of normal tissue irradiated was similar in both groups. Also, similar doses were delivered to the normal tissue. CONCLUSION: Differences in acute genitourinary toxicity are most likely secondary to radiation-induced prostatitis. Postprostatectomy radiation therapy is safe and is not associated with increased toxicity. PMID- 9205250 TI - Normal modiolus: CT appearance in patients with a large vestibular aqueduct. AB - PURPOSE: To determine the computed tomographic (CT) appearance of the normal modiolus and the pathologic alteration in patients with a large vestibular aqueduct and an otherwise normal-appearing cochlea. MATERIALS AND METHODS: Temporal bone CT studies obtained before and after a major upgrade of CT capability in 1992 were reviewed in four groups: Group A (1.5-mm section thickness) comprised 50 normal ears in 43 patients, group B (1-mm section thickness) comprised 75 normal ears in 50 patients, group C (1.5-mm section thickness) comprised 16 ears with a large vestibular aqueduct in 10 patients, and group D (1-mm section thickness) comprised 23 ears with a large vestibular aqueduct in 12 patients. All groups comprised adult and pediatric patients. RESULTS: In groups A and B, the normal modiolus was visualized in 90% and 100% of ears, respectively. In groups C and D, with a total of 39 ears with a large vestibular aqueduct and an otherwise normal cochlea, modiolar deficiency was demonstrated in 100% of ears. CONCLUSION: CT is an excellent technique for depicting the cochlear modiolus. Results suggest that all ears with a large vestibular aqueduct have associated cochlear modiolar deficiencies. Thus, a large vestibular aqueduct may be only occasionally, if ever, an isolated developmental anomaly of the inner ear. PMID- 9205251 TI - Primary hyperparathyroidism: higher success rate of first surgery after preoperative Tc-99m sestamibi-I-123 subtraction scanning. AB - PURPOSE: To evaluate the usefulness and cost-effectiveness of routine preoperative technetium-99m sestamibi-iodine-123 subtraction scanning in patients with parathyroid gland disease. MATERIALS AND METHODS: Tc-99m sestamibi-I-123 subtraction scanning was performed in 65 patients with primary hyperparathyroidism who were referred for evaluation before first surgery. RESULTS: Focal tracer uptake was detected in the mediastinum in two patients who then underwent primary sternotomy; a parathyroid adenoma, anterior to the ascending aorta, was resected in each case. In a third patient, imaging showed tracer uptake above the thyroid gland; this patient underwent resection of an undescended parathyroid adenoma located in the sheath of the right carotid artery. Initial surgery was curative in all patients. Preoperative subtraction scans depicted 56 of 59 (95%) solitary adenomas. Four patients had hyperplasia; two had double adenoma. Imaging findings indicated multiple parathyroid involvement in five of these patients and facilitated location of 12 of 15 (80%) enlarged glands. Four adenomas and two hyperplastic glands that weighed less than 100 mg were detected. The positive predictive value for any suspected location was 96%. Average surgery time was reduced from 120 to 90 minutes. CONCLUSION: Preoperative subtraction scanning is useful in planning parathyroid surgery and appears to be cost-effective. PMID- 9205252 TI - Carotid artery stenosis: external validity of the North American Symptomatic Carotid Endarterectomy Trial measurement method. AB - PURPOSE: To assess the generalizability of the North American Symptomatic Carotid Endarterectomy Trial method for determining the degree of stenosis on angiograms. MATERIALS AND METHODS: Six good-quality, baseline angiograms of carotid arteries that were less than 70% stenosed were reviewed by 14 experienced neuroradiologists at different academic institutions. All reviewers determined the degree of stenosis by calculating the ratio of the diameter of the artery at the point of maximal narrowing to the normal diameter distal to the stenosis (well beyond the carotid artery bulb). The reviewers marked the location of their measurements on the angiogram. Comparisons were performed among the reviewers' results and with the reference measurements. RESULTS: Interobserver agreement was 0.84 (95% confidence interval = 0.65, 0.97). The average interobserver disagreement of +/-7% was comparable with that reported in the literature. The overall bias was 6%, which indicated a tendency of the reviewers to overestimate the degree of stenosis in comparison with the reference determination. CONCLUSION: The North American Symptomatic Carotid Endarterectomy Trial reference measurements can be generalized beyond the bounds of this clinical trial, provided that attention is paid to details of the measurement method. PMID- 9205253 TI - Brain tumors: localized H-1 MR spectroscopy at 0.5 T. AB - PURPOSE: To investigate the feasibility of obtaining clinically useful magnetic resonance (MR) spectra at 0.5 T in cerebral lesions. MATERIALS AND METHODS: Single-voxel localized proton MR spectroscopy was performed at 0.5 T in 18 patients (aged 16-73 years) suspected of having cerebral lesions on MR images who subsequently underwent craniotomy and biopsy and in eight volunteers (aged 21-50 years). The metabolite resonances in the MR spectra were stratified according to the histologic diagnosis. MR spectra demonstrated resonances for choline containing compounds (Cho), creatine and phosphocreatine, glutamine and glutamate, N-acetylaspartate (NAA), myo-inositol, and lactate. RESULTS: In 14 of 15 patients with cerebral tumors, the Cho-NAA ratio was increased and was greater than 1. In three patients with nonneoplastic cerebral lesions, the ratio did not exceed 1. In healthy control subjects, the average ratio was 0.54. CONCLUSION: H 1 MR spectroscopy at 0.5 T provides clinically useful information in patients with cerebral lesions. PMID- 9205254 TI - Brain abscess and brain tumor: discrimination with in vivo H-1 MR spectroscopy. AB - PURPOSE: To determine the ability to differentiate brain abscess from cystic or necrotic brain tumor with hydrogen-1 magnetic resonance (MR) spectroscopy. MATERIALS AND METHODS: H-1 MR spectroscopy was prospectively performed in seven consecutive patients with pyogenic brain abscess and in seven consecutive patients with necrotic or cystic brain tumor (five patients with glioblastoma and one each with pilocytic astrocytoma and metastasis from lung cancer) in whom radiologic images depicted ring-shaped areas of contrast material enhancement (indicative of a cystic or necrotic mass). Assignment of resonance peaks to metabolites was based on reports in the literature. RESULTS: In six of seven patients with abscess, there were various resonances attributed to lactate, valine, alanine, leucine, acetate, succinate, and unidentified metabolites (2.2, 2.9, 3.2, 3.4, and 3.8 ppm). In six of seven patients with tumor, there was only a resonance attributed to lactate. One patient with a tumor had an unidentified peak at 0.9 ppm (presumably attributed to lipid) in addition to the peak attributed to lactate. CONCLUSION: Spectral patterns from in vivo H-1 MR spectroscopy may permit differentiation of brain abscess from necrotic or cystic tumor. PMID- 9205256 TI - Secondary thalamic degeneration after cerebral infarction in the middle cerebral artery distribution: evaluation with MR imaging. AB - PURPOSE: To evaluate secondary degeneration of the ipsilateral thalamus after cerebral infarction in the middle cerebral artery (MCA) distribution by using magnetic resonance (MR) imaging. MATERIALS AND METHODS: Thirty patients (17 men, 13 women; aged 30-85 years) with embolic cerebral infarction in the MCA distribution underwent serial MR imaging 2 hours to 12 months after a stroke. In 23 of the 30 patients, the authors evaluated cerebral blood flow with single photon emission computed tomography (SPECT). RESULTS: T2-weighted spin-echo images disclosed a hyperintense area in the ipsilateral thalamus in 14 patients (47%) 1-12 months after stroke. The hyperintense area was confined to the dorsomedial or anterior and dorsomedial nuclei of the thalamus in nine of the 14 patients; it extended from the dorsomedial or anterior and dorsomedial nuclei to the ventral lateral nucleus or pulvinar in the remaining five patients. Hypoperfusion of the ipsilateral thalamus was observed in 21 of the 23 patients who underwent SPECT. Twelve of the 21 patients also showed a hyperintense area in the ipsilateral thalamus on the MR images. CONCLUSION: MR imaging is useful in evaluating secondary thalamic degeneration after cerebral infarction. In clinical practice, this secondary degeneration should not be mistaken for other lesions such as further cerebral infarction. PMID- 9205255 TI - Thermal effects of focused ultrasound on the brain: determination with MR imaging. AB - PURPOSE: To determine the feasibility of the use of temperature-sensitive magnetic resonance (MR) imaging for the detection of local temperature elevations at the focus of a low-power ultrasound beam in the brain. MATERIALS AND METHODS: The brains in 28 rabbits were sonicated at acoustic power levels of 3.5-17.5 W. Four to five different locations were sonicated at different acoustic power levels in each rabbit. MR images were obtained 2 hours, 48 hours, 10 days, and 23 days after the sonications, depending on when the animals were sacrificed. Histologic evaluation of whole brain was performed. RESULTS: Forty of 43 (93%) of the lowest-power (3.5-W) sonications were visible on temperature-sensitive MR images and did not result in any short- or long-term histologic or MR imaging evidence of tissue damage. A contrast-to-noise ratio of approximately 6 and a temperature elevation of 7 degrees-8 degrees C were observed. CONCLUSION: Temperature elevations induced by means of focused ultrasound exposures that do not cause damage in the in vivo rabbit brain can be detected at temperature sensitive MR imaging. PMID- 9205257 TI - In vivo target-specific delivery of macromolecular agents with MR-guided focused ultrasound. AB - Magnetic resonance (MR) images in rabbit muscle were used to select a target region for application of pulsed-focused ultrasound. Liposomes encapsulating gadopentetate dimeglumine, an MR-detectable model representing pharmaceutical agents, were then injected intravenously. Focal enhancement on T1-weighted images, representing gadolinium-liposome accumulation, occurred in the pulsed focused-ultrasound focal zone. Therefore, MR-guided pulsed-focused ultrasound targeted the delivery of macromolecular pharmaceutical agents within tissues. PMID- 9205258 TI - Internally stabilized spine: optimal choice of frequency-encoding gradient direction during MR imaging minimizes susceptibility artifact from titanium vertebral body screws. AB - In 18 vertebral bodies with titanium fixation screws and in a phantom model, visualization of the vertebral body marrow was improved and susceptibility artifact was reduced on T1-weighted spin-echo magnetic resonance images when the direction of the frequency-encoding gradient was parallel to the long axis of the screw. A perpendicular direction improved image quality only when the region of interest was adjacent to the tip of the screw. In the phantom, the length of the screw was statistically significantly increased and the width and area were reduced (P <.001) when the gradient was parallel to the long axis of the screw. PMID- 9205259 TI - Small vessels in the human brain: MR venography with deoxyhemoglobin as an intrinsic contrast agent. AB - To assess a magnetic resonance (MR) imaging method for depicting small veins in the brain, a three-dimensional, long echo time, gradient-echo sequence that depended on the paramagnetic property of deoxyhemoglobin was used. Veins with diameters smaller than a pixel were depicted. This MR imaging method is easy to implement and may prove helpful in the evaluation of venous diseases. PMID- 9205260 TI - Iterative image reconstruction reduces apparent tumor distortion. PMID- 9205261 TI - Can relative contrast agent concentration be measured in vivo with color Doppler US? PMID- 9205263 TI - Sleep apnea syndrome: evaluation with dynamic MR imaging of the upper airway. PMID- 9205264 TI - PET data analysis of pulmonary abnormalities. PMID- 9205265 TI - Lung nodule enhancement at CT. PMID- 9205267 TI - Women find their voices. The success of community outreach and case finding. AB - Families in transition (FIT), a joint program between Beth Israel Medical Center (BIMC) and National Development and Research Institutes, Inc., uses indigenous neighborhood women, Companeras (companions), to conduct street outreach in a New York City neighborhood where HIV infection is rampant. The outreach workers inform people that, not only will they be assisted in talking about HIV and guardianship for their children, but that they will be provided information concerning access to food stamps, housing, health care, and other basic needs. Through community participation, the Companeras empower themselves through knowledge making and sharing and, in the process, create conditions in which other women can began to effect changes in their individual lives and families. The ultimate goal of the project is to support HIV-infected women in finding their voices so that they can talk with their children and plan for their futures. PMID- 9205268 TI - A model emerges for the community-based nurse care management of older adults. AB - Changes in the demographics of older adults and the structure and financing of the health care delivery system have created a need to develop alternative models of care delivery for the elderly. The nurse care management model is a potentially cost-effective solution for provision of comprehensive care to this population. By providing timely health promotion and illness prevention education, as well as coordinating community resources, nurses can reduce the health care costs of this growing segment of the population. Funding this model, however, remains a challenge as such services are not directly reimbursed by third-party payers. PMID- 9205269 TI - Women speak. Healing the wounds of homelessness through writing. AB - The Women Speak writing project explores the use of writing as a therapeutic process with homeless women at an urban drop-in center. The project was developed in response to the women's desire to give voice to their experiences. By speaking about and sharing the experiences of their lives, a sense of empowerment through expression began. Nursing students and faculty working at the drop-in center were challenged to re-think the traditional clinical site relationship which gives highest priority to meeting the needs of students and faculty. PMID- 9205270 TI - Retirement communities as creative clinical opportunities. AB - The extraordinary number of elderly expected to live into the next century calls for creative inclusion of this population in learning experiences for students in baccalaureate nursing programs. Retirement communities offer a rich environment for interaction and learning with the well elderly that encourages students to consider the limitless possibilities for nursing interventions with our greatest national resources: the elderly. PMID- 9205271 TI - Missouri responds to the advanced practice nurse role. AB - A randomly-drawn statewide sample of 891 consumers revealed overall support for the advanced practice nurse role to be greater than 75 percent. Seeking health care consumers' reactions to proposed alternatives is a crucial step in planning and implementing a program of health care reform that will meet current and future health needs. PMID- 9205272 TI - Art in practice. The New Leaf Project. PMID- 9205273 TI - We still have much to do. PMID- 9205274 TI - Decline in nursing school enrollment continues according to latest NLN study. PMID- 9205275 TI - Associate degree nursing programs accredited by the NLN 1997-98. PMID- 9205277 TI - Necrotizing fasciitis: a case study. AB - Necrotizing fasciitis (NF) is a disease characterized by the destruction of the skin, subcutaneous fat, and fascia, with or without inflammation of the muscle tissue. NF carries a mortality rate in excess of 50 percent in neonates. Rapid diagnosis and initiation of antibiotics, along with meticulous nursing care, can provide an acceptable outcome for the neonate. This article reviews the pathophysiology, diagnosis, and treatment of NF; presents a case study; and provides a nursing care plan that supports treatment of this disease process. PMID- 9205276 TI - Persistent pulmonary hypertension of the newborn: case study and pathophysiology review. AB - Pulmonary hypertension, though quite normal in the fetus while the burden of oxygenation is on the maternal placenta, can be rapidly fatal if severe resistance to pulmonary blood flow continues as the infant attempts to make the transition to extrauterine life. This article reviews the pathophysiology of and current management priorities for persistent pulmonary hypertension of the newborn. PMID- 9205278 TI - Variables associated with parental stress in neonatal intensive care units. AB - A descriptive study was conducted to identify sources of parental stress in two types of NICUs. Data were collected from 212 parents and a single interview was done within three weeks of the infant's admission. Parents completed the Parental Stressor Scale: NICU, Spielberger State-Trait Anxiety Inventory, Life Events Scale, and Parent Questionnaire. Data extracted from the infant's chart were used to complete the Neonatal Morbidity Scale and Baby Data Sheet. Data were analyzed using multiple regression techniques. Findings indicate that mothers and fathers differed in their responses to this experience. How parents perceived the severity of their infant's illness was the most powerful variable associated with their stress scores. Trait anxiety, desire for the pregnancy, and where and when parents first saw the baby were other variables significantly correlated with stress scores. PMID- 9205279 TI - Cardiac embryology. AB - The cardiovascular system is the first major organ system to develop in the embryo, being fully formed and functioning by the eighth week of gestation. This early, rapid development may explain the prevalence of congenital heart disease. Infants with congenital heart disease require considerable nursing skill and an understanding of the particular diagnosis. Nurses who understand basic cardiac embryology will better understand the various defects, how they occur, and how they affect cardiac physiology and functioning. This article discusses development of the heart and major vessels arising from the heart and relates stages of development to corresponding congenital heart defects. PMID- 9205280 TI - Determining the relationship between invasive and noninvasive blood pressure values. AB - Blood pressure measurements are a common indicator of cardiovascular status. The two most frequently used methods of obtaining this measurement in the NICU are direct measurement using an umbilical arterial catheter and indirect measurement with a cuff. Cuff values obtained from the upper arm or thigh are reported to correlate with umbilical arterial measurements. It is common practice in some NICUs to utilize the calf of the infant's leg to obtain cuff blood pressure readings. The goal of this pilot investigation was to determine if blood pressure values obtained from the left calf of neonates weighing at least 800 gm correlate with measurements obtained from the umbilical arterial catheter. A descriptive correlational design was used for this investigation. A convenience sample of 20 infants was obtained. Data were analyzed using the Pearson r with a significance level of .01. PMID- 9205282 TI - Isolation techniques. PMID- 9205283 TI - Interpretation of fetal cord blood gases. PMID- 9205284 TI - A stabilization technique for endotracheal tubes in premature infants. PMID- 9205285 TI - A cleaner blood sampling apparatus. PMID- 9205286 TI - Circles of influence. PMID- 9205287 TI - Time is of the essence. PMID- 9205288 TI - Countersigning notes by others. PMID- 9205289 TI - Earning CE credits on the Net. PMID- 9205290 TI - Taking another route to pain relief. PMID- 9205291 TI - Safely administering a blood transfusion. PMID- 9205293 TI - Actionstat. Rattlesnake bite. PMID- 9205292 TI - Incorrect restraint use. PMID- 9205294 TI - New drugs Part II. PMID- 9205295 TI - 26 words toward a successful nursing career. PMID- 9205296 TI - Eleanor walled herself in. PMID- 9205297 TI - Caring for patients with non-Hodgkins lymphoma. PMID- 9205298 TI - Bradycardia. Keeping the current flowing. PMID- 9205299 TI - Using a peak flowmeter. Monitoring the air waves. PMID- 9205300 TI - Learning about low-profile gastrostomy devices. PMID- 9205301 TI - What a travel nurse company seeks in you. PMID- 9205302 TI - What you'll need on your first day. PMID- 9205303 TI - Things remembered. PMID- 9205304 TI - 6 safety suggestions. PMID- 9205305 TI - I.v. rounds. Dealing with infiltration. PMID- 9205306 TI - Sharing. The list. PMID- 9205307 TI - Is the case manager being replaced? PMID- 9205308 TI - The promise of automated wellness systems. AB - Do automated wellness programs hold out the promise of improving the health of individuals and populations through the establishment of baseline wellness checkpoints? Nurse case managers have a strong interest in this question. The automation of wellness programs and the integration of this information into the continuum of a care information system model offers ongoing, interactive accountability of individual health status. The crowning glory will be the ultimate integration of the individual's personal health game plan with the provider's continuum of care offering. The strength of the nursing profession lies in its traditional role of promoting health through preventive measures (public health nursing) and managing the ill patient's progress toward his/her optimal health status. Therefore, nurses and nurse case managers should embrace wellness programs and wellness data availability. Because of their experience in using and understanding wellness data, nurses are in a unique position. Not only will they be able to assimilate rapidly the use of wellness data into their own practice, they also will be able to guide other professionals in its use. The table in this articles provides examples of the impact that automated wellness program data could have at various stages of the clinical decision-making process. PMID- 9205309 TI - Structured care methodologies: tools for standardization and outcomes measurement. AB - In today's healthcare environment, institutions are striving to streamline processes, reduce costs of healthcare, and establish best practice patterns while maintaining and improving the quality of care provided. Various healthcare delivery models are in use including case management and outcomes management. Various tools or structured-care methodologies (SCMs) are incorporated into these different models to support cost reduction and streamline processes while enhancing quality of care. This article discusses the tools frequently used, such as critical pathways, algorithms, and guidelines, as well as how these tools can be used in combination to support each other. This article also addresses the benefits of SCMs, how these tools are developed, and how the data obtained can be used in quality enhancement programs. PMID- 9205310 TI - Negotiating the chaos of patient care with clinical pathways. PMID- 9205311 TI - Pneumonia pathway streamlines care. PMID- 9205312 TI - Discharge dilemma: planning for the out-of-state resident. PMID- 9205314 TI - Reward and recognition. PMID- 9205313 TI - Assessing clinical path effectiveness: a model for evaluation. AB - Clinical paths are being developed in response to the current push to decrease healthcare costs while maintaining high quality care. Literature supports clinical path evaluation results, but few articles address a systematic evaluation process. The purpose of this article is to present a five-step evaluation model that can be followed in determining the effectiveness of clinical paths. The goal of the evaluation process is to provide reliable information and to translate the information into changes in healthcare. The article contains a graphic representation of the model, an explanation of each step, and activities to be performed. PMID- 9205315 TI - Quality problem-solving, decision-making, type theory, and case managers. AB - Temperament theory can serve as a tool to improve the case manager's effectiveness. This knowledge identifies strengths and liabilities that can enhance individual and group problem-solving, decision making and team productivity. This article will examine components of decision making, temperament or personality theory, and describe major types. Part II will discuss the implications of typewatching for case managers and others with whom they interact with respect to time management, negotiation, and team building. off PMID- 9205316 TI - The Internet: a knowledge source for nurse case managers. PMID- 9205317 TI - Initiatives to build a competitive healthcare system: integrating service redesign and clinical pathways. AB - In this article, the authors describe how two separate initiatives (the redesign of hospital services and the development and implementation of clinical pathways) were integrated at an academic medical center. The lessons learned in integrating projects that were not conceived at the same time and in developing multidisciplinary teams to produce new case management tools may be used by clinicians and managers elsewhere. The service redesign project developed a role for senior, experienced nurses with clinical expertise and patient management skills to manage the care of groups of similar patients, focusing on patient outcomes. The clinical pathway project developed diagnostic related groups-based clinical pathways. The steps involved in clinical pathway development-stakeholder cooperation building, initiation of data collection tools, data analysis, implementation of the role of the expert nurse in "championing" the pathway, and evaluation of the effects of pathways-are discussed. PMID- 9205318 TI - Case management's role in human immunodeficiency virus care. PMID- 9205319 TI - Integration of the clinical nurse specialist into the nurse case manager role. AB - The focus of healthcare is changing from treatment of illness to promoting optimal wellness in a cost-effective manner. With this change, the role of the nurse also must change. The traditional role of the clinical nurse specialist (CNS) is being eliminated from many hospitals, leaving a gap is nursing expertise. The role of the nurse case manager (NCM) is developing rapidly. A logical movement is for the CNS to move into the NCM role. Because of distinct differences between these roles, the CNS must learn new skills. This paper discusses the role differences between the NCM and the CNS and suggests areas of education needed to make a successful transition. But more importantly than just learning new skills, the CNS must undergo a transformation in perspective to function in the new reality of managed care. PMID- 9205320 TI - Subacute case management: carpe diem. PMID- 9205321 TI - Ready, set, go.... PMID- 9205322 TI - Case management in a wound care program. AB - Patients who need wound care can present a complex challenge in the current managed care environment. Managing health care benefits and financial and socioeconomic factors that impact wound healing are as crucial to wound healing as providing aggressive wound care. Case management is used to describe the strategies involved in coordinating patient care to achieve optimal clinical and financial outcomes and ensure quality and continuity of care. In this article, the author illustrates how case management plays an integral part in a wound care program. The article features tips on how to effectively use limited financial resources that burden many patients who need wound care, includes ways to approach physicians when current wound care is not effective, and focuses on early discharge planning and a multidisciplinary team approach to ensure optimal clinical and financial outcomes. To help sharpen skills, included at the end of the article is a Case Study Challenge and tools to use in practice: Case Management Plans for Wound Patients. PMID- 9205323 TI - Professional presentations: a primer for case managers. PMID- 9205324 TI - Evolution of a role to enhance care coordination. AB - The recent trend toward shorter hospital stays has prompted health care personnel in acute care settings to reevaluate models of coordinating patient care. In whatever model is adopted, there must be someone whose role is to focus on achieving quality patient outcomes in a shorter time frame. Our institution has piloted several different models with various job classes to fulfill the role. However, the primary focus of the models-to enhance care coordination-remained unchanged. The initial care coordination model was the development of patient care coordinator role by converting the charge nurse position on the unit. The second model our institution developed occurred during a management restructuring. This model created a new position, a nursing practice coordinator, who had previously been a management support person with supervisory responsibilities. We now envision the role to be assumed by a nurse practitioner. The nurse practitioner's broader scope of practice potentially can further enhance coordination of care. In this article, the authors describe the development process, issues and interventions, and lessons learned during each step of redesigning a case management model. PMID- 9205325 TI - Critical pathway and patient and family teaching protocol for major depression. AB - The pressures to contain cost, reduce resource consumption, and articulate outcomes will continue in psychiatric-mental health care. Case management and critical pathways focused on outcome management are systems and tools used to provide care in a cost-effective manner while maintaining the quality of patient care. PMID- 9205326 TI - Outcomes system implementation for subacute care. AB - The emergence of subacute care, seen as a cost-effective alternative to other, more expensive settings, is an important option for case managers. The current, rapid growth of subacute care, the diversity of subacute programs, the differences in patient types, and the lack of consistent standards to define subacute care illustrate the critical need for the case manager's role in balancing quality and cost. The lack of solid clinical outcomes data for patients treated in subacute care make it difficult for case managers to assess quality. Outcomes data, as a measurable dimension of quality, may include clinical effectiveness measures, associated costs, and patient/family satisfaction. Outcomes data can be used as a tool by the case manager to facilitate the coordination of patient care. Although there is an increased consumer interest in outcomes across all health care modalities, there is limited outcomes research available to document the efficacy of subacute care. As the pressure increases for outcomes data on subacute care by consumers and payers, efforts toward facility implementation of outcomes systems to assess subacute care are growing. The unique challenges to outcomes implementation in a skilled nursing facility based setting are discussed, and strategies for successful implementation are presented. Basic subacute outcomes implementation issues of organizational support, staff participation, and data collection are reviewed. Ideas for case management involvement with facility implementation are discussed. PMID- 9205327 TI - The NHS executive's new guidelines on violence in A&E departments. PMID- 9205329 TI - Local pay not yet buried. PMID- 9205328 TI - Check and check again. PMID- 9205330 TI - The other NHS. PMID- 9205331 TI - Capital punishment. PMID- 9205332 TI - Takeaway nappies. PMID- 9205333 TI - Maxi-nurses. PMID- 9205334 TI - Mind the gap. PMID- 9205335 TI - Leading from the front. PMID- 9205336 TI - New horizons. PMID- 9205337 TI - The misery of hay fever. PMID- 9205338 TI - Reflections for action. AB - This is the second of three articles based on a major report launched by the Foundation of Nursing Studies last year-Reflection for Action-which looked at the dissemination and implementation of research in nursing. This article discusses two of the studies that made up the report. PMID- 9205339 TI - Alleviating pre-operative anxiety in patients: a study. AB - This study aimed to provide an objective view of the relationship between the giving of information, anxiety and hospital admission. Previous research has suggested that patients are already anxious on admission to hospital and that any information given at this time may be forgotten easily or misunderstood. Forty patients listed for simple elective surgery participated in this study. Using an experimental design, subjects in the experimental group were interviewed and information given to them in their own home before admission and again on their first day in hospital. A state-trait anxiety questionnaire was used to compare both groups. The resultant difference in the anxiety level shown between both groups on admission was found to be significant. PMID- 9205340 TI - Community nursing profiles: their role in needs assessment. AB - The last article in our series of three looking at public health nursing explores the background from which community nursing profiles have developed into annual activities in provider trusts around the country. The author suggests that there are important issues which prevent them from being a valuable resource for needs assessment and service planning. PMID- 9205341 TI - Working with people. PMID- 9205342 TI - Dobson's choice. PMID- 9205343 TI - Shedding light on a system under strain. PMID- 9205345 TI - Chaos, exploitation and heartache for enrolled nurses. PMID- 9205344 TI - Should they stay... or should they go? PMID- 9205346 TI - Enrolled and endangered. PMID- 9205347 TI - Spread the world. Interview by Jane Salvage. PMID- 9205348 TI - Friends for life. Interview by Eileen Fursland. PMID- 9205350 TI - A bad press for nurses? PMID- 9205349 TI - Who are you calling paranoid? PMID- 9205351 TI - Shiny, happy people just doing what comes naturally. PMID- 9205352 TI - Those lazy, hazy, crazy days of summer. PMID- 9205354 TI - Travellers' ills. PMID- 9205353 TI - A healthier shade. PMID- 9205355 TI - The vote race. PMID- 9205356 TI - Lucky dip. PMID- 9205357 TI - A change for the better. PMID- 9205358 TI - How to make a questionnaire work. AB - This article is the final one in a series on compiling a questionnaire, and it examines layout, issues of reliability and validity and piloting. PMID- 9205359 TI - Growth hormone deficiency in adults. AB - This article describes growth hormone deficiency in adults. The causes and clinical features are explained and the article looks at treatment approaches, their effects and the role of the nurse. PMID- 9205360 TI - A positive approach to chemotherapy. AB - This article deals with the care of patients receiving chemotherapy for breast cancer. Using a case study, it highlights the importance of supporting patients and helping them to cope with their fears and anxieties about the treatment and its possible side-effects by considering not only their physical but also their psychological needs. PMID- 9205361 TI - Under surveillance. PMID- 9205362 TI - A treatment of last resort. PMID- 9205364 TI - Making a fresh start. PMID- 9205363 TI - Know how. Pulsating air suspension therapy. AB - The realisation that pressure sores are caused by shearing forces and tissue distortion rather than simply by pressure, had led to the development of specialised patient support systems. The pulsating air suspension system was developed to help reduce the incidence of oedema formation. All health-care professionals using specialised bed therapy have a responsibility to understand the principles of that therapy in order to ensure appropriate patient selection, which may be aided by the development of placement criteria. The concept of pulsation therapy has been a stimulus for new thinking on the causes and treatment of pressure sores and has highlighted the need for further research on the part played by the lymphatic system and oedema on pressure sore formation, although there is already strong evidence that there is a link between tge two. PMID- 9205365 TI - Assessing the future. AB - Awareness of the importance of wound assessment and management appears to be increasing, and centres of excellence are emerging. Nurses have a responsibility to provide effective research-based care for patients, but require knowledge and training to do this. Research-based practice is important, but there is a case for more nurses being involved in research in this area. Nurses realise that, where disagreement in wound management arises between themselves and other professions, one way to overcome this is by demonstrating that they are skilled, proficient and knowledgeable practitioners in wound care. In this way, autonomy may be promoted and nurses can continue to push forward the boundaries of wound management, questioning rituals and promoting innovation. Good documentation is also important and the use of a wound assessment tool may assist in this. The challenge now is for nurses to demand adequate training in order to help them become the skilled, proficient and knowledgeable practitioners they strive to be. PMID- 9205366 TI - Frank Dobson tough line on tobacco sponsorship. PMID- 9205367 TI - Nursing careers founder 'on a broken ladder'. PMID- 9205368 TI - There's a will but is there a way? PMID- 9205369 TI - Buy privately now and we will all pay later. PMID- 9205370 TI - Labour faces the tricky task of retaining the traditional face of the NHS while attracting private finance. PMID- 9205371 TI - Know how. Treatment of acne. AB - Acne is a common condition, affecting most adolescents, although for the majority it subsides within five years. For some, however, it causes great distress: they feel unattractive, isolated and self-conscious, sometimes developing psychological problems. These people should be encouraged to seek help from their GP, as treatment is available. PMID- 9205372 TI - Lost children. Who cares. PMID- 9205373 TI - Care and control. PMID- 9205374 TI - Exploring schizophrenia. PMID- 9205375 TI - In search of the right balance. PMID- 9205376 TI - The relentless army. PMID- 9205377 TI - Dying to make a profit. PMID- 9205379 TI - Debt Control. PMID- 9205378 TI - Beer & Fags & nursing. Interview by Richard Morris. PMID- 9205380 TI - It is foolish to even think about researching the effectiveness of clinical supervision before it has been fully implemented. PMID- 9205381 TI - Teenage suicide attempts. AB - This article describes the care of adolescents with suicidal intentions or after a suicide attempt. The special role of the nurse in creating a relationship with the patient and providing a role model for the family is examined. PMID- 9205382 TI - What are the best ways of tackling obesity? AB - A systematic review of the literature on the treatment and prevention of obesity carried out by the NHS Centre for Reviews and Dissemination (CRD) demonstrates that the prevalence of excess weight and obesity are on the increase. The review indicates that certain interventions in primary care can be effective, while surgery appears to be most effective in severe obesity. It highlights limitations in the available research in the area. The article is abstracted from the April 1997 issue of Effective Health Care published by the NHS CRD. PMID- 9205383 TI - Wax, sunlight and X-rays. PMID- 9205384 TI - The social effects of juvenile chronic arthritis. AB - We are continually expanding our knowledge of JCA and establishing our understanding on an ever-increasing knowledge base encompassing immunology, immunogenetics, physiology, psychology and sociology. The causes and treatment of this condition are multifactoral and no one discipline can hope to embrace all of them. Nurses, particularly those with rheumatological and/or paediatric expertise, are uniquely placed to provide the link between patient, the family and health care team and thus to coordinate treatment and ensure that care meets the needs of youngsters with JCA and their families. PMID- 9205385 TI - Home economics. PMID- 9205386 TI - New kids on the block. PMID- 9205387 TI - Birds-eye view. PMID- 9205388 TI - Recommendations for using reverse staging to complete the MDS-2. AB - The use of the Minimum Data Set (MDS) for documentation in long term care has resulted in the widespread reliance on reverse staging to document the healing of pressure and venous ulcers. This article presents a pragmatic approach for defining the healing process in a staging nomenclature so that it is physiologically accurate. PMID- 9205390 TI - To culture, or not ... and if yes, how? PMID- 9205389 TI - A comparison of two culturing methods for chronic wounds. AB - Bacterial infection has always been a potential complication of any wound. Controversy exists regarding the significance of bacteria in chronic wounds. It is important to accurately diagnose wound infection by bacterial identification and quantification in order to prevent unnecessary and/or inappropriate treatments and to minimize patient complications. The primary function of culturing is to identify infection in a wound. The tissue culture is an accepted standard for measuring infection, although swab cultures are commonplace in the clinical setting. The purpose of this study was to test the differences in bacterial counts and identification in swab and tissue cultures taken from the same wound site of 10 chronic wounds. It was hypothesized that if swab and tissue cultures are equally effective in identifying and quantifying the organisms in a chronic wound, they are equally effective methods in determining infection in the chronic wound. The reliability, validity and limitations of the study are discussed, as well as the statistical analyses and results. PMID- 9205391 TI - Culturing wounds in clinical practice. PMID- 9205392 TI - Comments on statistical validity. PMID- 9205393 TI - Pressure ulcer prevalence and prevention of nosocomial development: one hospital's experience. AB - A 500 bed acute care facility needed to replace their old medical-surgical patient beds and reduce the costly use of specialty beds and overlays. After a review of the literature, the facility focused on the 44-bed AIDS/Oncology unit, first trialing a new therapeutic bed, then replacing all the beds. A prevalence survey was conducted 5 days before the placement of the new beds and monthly thereafter for 6 months. It was hypothesized that (1) incidence of nosocomial pressure ulcers would decrease, (2) use of specialty beds would be reduced, resulting in significant cost savings within a few months, and (3) there would be a learning curve regarding use of the beds and proper "zoning" of patients. From April through November 1995, 256 patients were surveyed. Pre-survey, the average range on the unit for pressure ulcer prevalence was 7.5 to 15% (both nosocomial and admitted). Post-survey, the range was 3 to 16% (admitted ulcers only). Zero nosocomial pressure ulcers developed during the study period. Use of foam overlays and low air loss surfaces decreased, resulting in a savings of 83%. There were no problems with using the beds or zoning patients. These survey results suggest that other institutions could achieve similar clinical and financial outcomes by converting rental dollars to capital assets. PMID- 9205395 TI - Stoma site marking: a primer. AB - The purpose of pre-operative stoma site marking is to select an appropriate location in an area of the abdomen for surgical placement of a stoma. A poorly located stoma may result in pouching problems, increase potential for leakage, and place undue hardship and emotional trauma on the patient. The type of ostomy surgery must be known to select the appropriate abdominal quadrant to be marked. The ideal stoma site is located below the umbilicus, within the rectus muscle, away from scars, creases, bony prominences, umbilicus, and belt line, on the summit of the infraumbilical fat mound, and visible to the patient. It is vital for the clinician to be aware of any extenuating circumstances of the patient that may alter the site selection. Optimal stoma site placement will promote self care and rehabilitation of the patient. PMID- 9205396 TI - Practical guidelines for developing a hospital-based wound and ostomy clinic. AB - Multidisciplinary teams save time and money while improving quality of patient care. Hospital-based wound/ostomy clinics provide patients with holistic, one stop, expert care with access to a variety of diagnostic and treatment modalities. The authors discuss how to assemble all the ingredients for a successful multidisciplinary hospital-based clinic, how to make it work, and how to measure its cost-effectiveness for patient, hospital, community and nation. The original model clinic established in Costa Rica using these methods was so successful at providing efficient, high quality, cost-effective care that it has been adopted in 17 different locations there, and similar clinics are being developed in other countries. This model is applicable to industrialized and developing countries. By focusing hospital resources on evidence-based wound/ostomy care, it has the potential to revolutionize wound and ostomy care around the world. PMID- 9205397 TI - Pressure ulcers: collaboration in wound care. Is there a reasonable approach? AB - Pressure ulcers represent a significant impact on utilization of healthcare beds, cost for the insurer, and adverse emotional impact for the family. With this in mind, one must address in an effective fashion a method of objective assessment using the current methodology to deal with risk factors in mobility, disease states, and nutrition, as well as give significant attention to prevention protocols. One should adhere to a schedule of turning the patient, sleep surfaces, and appropriate skin care while recognizing the impact of each. Finally, treatment modalities should be undertaken, not only with cost-effective issues in mind, but with ease and convenience for the healthcare provider and the least discomfort for the patient as well. One can focus on debridement by way of enzymatic, autolytic, or surgical methods, recognizing surgery as the least effective for our goals in the management of this patient. Assessment and, finally, reassessment using the available scales will allow us to provide effective healthcare in a therapeutic fashion. PMID- 9205398 TI - Managing conflict in the nursing home environment. PMID- 9205399 TI - Outcomes documentation to support contracts and payment. PMID- 9205400 TI - Advanced Tissue Sciences and Smith & Nephew present Dermagraft data. PMID- 9205401 TI - Ethics in action. Contract negotiations at your hospital. PMID- 9205402 TI - A dialysis alternative more nurses can run. PMID- 9205403 TI - Elective Cardioversion. Who, when, and how. PMID- 9205404 TI - When you want humidity. PMID- 9205405 TI - Reverence. PMID- 9205406 TI - Revelation. PMID- 9205408 TI - Look at the product side. PMID- 9205407 TI - Pharmacology in practice: Antiarrhythmics. PMID- 9205409 TI - HIV and the law. PMID- 9205410 TI - Why is this patient suffering jaw pain? PMID- 9205412 TI - The antibody response in human autoimmune thyroid disease. AB - 1. The analysis of the antibody response in autoimmune thyroid disease has followed several historical trends. It was the investigation of thyroid-reactive antibody that allowed the initial characterization of the three principle thyroid autoantigens, thyroglobulin, thyroid peroxidase and the thyroid stimulating hormone receptor. 2. Analysis can be grouped under two broad areas: analysis of the physiological and pathological effects of the antibody, and analysis of the structure of the antibodies themselves. This review will focus on the latter. 3. Within recent years there has been a great increase in knowledge of thyroid reactive antibody structure, principally through the adoption of phage display combinatorial library methodologies. While this latter technique has established some general principles for antibodies to thyroglobin and especially thyroid peroxidase, there is still a substantial gap in our knowledge of the antibody response to the thyroid stimulating hormone receptor. 4. Thyroid peroxidase antibodies have a relatively restricted V-region usage, and there is a correlation between the V-regions used and the epitope on thyroid peroxidase bound. In particular the V kappa light chain V kappa I(O12), is associated with reactivity to one epitope. 5. The purpose of this review is to bring together the latest results concerning the molecular analysis of the antibody response in autoimmune thyroid disease, to highlight areas of ignorance and conflict, and to discuss the methods adopted to circumvent the problems associated with analysis of the antibody response. PMID- 9205413 TI - Fractal component of variability of heart rate and systolic blood pressure in congestive heart failure. AB - 1. There is a substantial non-harmonic or fractal component to the variability of both heart rate and blood pressure in normal subjects. Heart rate is the more complex of these two signals, with respect to the slope, beta, of the 1/f beta relationship. In congestive heart failure, heart rate spectral power is attenuated, but the fractal and harmonic components of heart rate and systolic blood pressure variability have not been characterized. 2. Two groups, each comprising 20 men, were studied during 15 min of supine rest and spontaneous respiration: one with functional class II-IV heart failure (age 52 +/- 2 years; mean +/- SEM) and a second group of healthy men (age 46 +/- 2 years). 3. Total spectral power for heart rate was significantly reduced in heart failure (P < 0.02), whereas total spectral power for systolic blood pressure was similar in the two groups. In both heart failure and normal subjects, 65-80% of total spectral power in these two signals displayed fractal characteristics. 4. In heart failure, the slope of the 1/f beta relationship for heart rate was significantly steeper than in normal subjects (1.40 +/- 0.08 compared with 1.14 +/- 0.05; P < 0.05), indicating reduced complexity of the fractal component of heart rate variability. There was no significant difference in the 1/f beta slope for systolic blood pressure variability between these two groups, but the blood pressure signals were less complex than heart rate variations in both heart failure (2.31 +/- 0.15; P < 0.006) and normal subjects (2.47 +/- 0.15; P < 0.0001). 5. Parasympathetic nervous system activity, as estimated from heart rate variability was reduced (P < 0.01) in patients with heart failure, whereas trends towards increased sympathetic nervous system activity and decreased non-harmonic power were not significant. 6. The non-harmonic components of cardiac frequency are reduced in heart failure. Non-harmonic power is not attenuated, but the complexity of the heart rate signal is less than in subjects with normal ventricular function. A reduction in parasympathetic modulation appears to contribute to this loss of complexity of heart rate. Consequently, the heart rate signal comes to resemble that of blood pressure. In contrast, the variability and complexity of the systolic blood pressure signal is similar in heart failure and normal subjects. This reduced complexity of heart rate variability may have adverse implications for patients with heart failure. PMID- 9205414 TI - Blood pressure parameters as determinants of small artery structure in human essential hypertension. AB - 1. Adaptive changes in small arteries may be more closely correlated with pulse pressure than with systolic, diastolic or mean blood pressures in human essential hypertension. 2. An analysis was performed on the structure of small arteries, age and blood pressure measurements obtained from 56 patients with untreated essential hypertension and 56 matched normotensive volunteers to examine the association between age, blood pressure and small artery structural parameters. 3. Essential hypertension was associated with an increase in media thickness and a decrease in lumen diameter, resulting in an increase in media/lumen ratio. 4. There was a significant correlation between age and media/lumen ratio in normotensive volunteers but not in patients with essential hypertension. 5. There was no correlation between any blood pressure and structural parameter in normotensive volunteers. 6. Both diastolic and mean blood pressures in essential hypertension correlated with media/lumen ratio (P < 0.01); systolic blood pressure correlated less well (P < 0.02). However, pulse pressure did not correlate with media/lumen ratio, suggesting that it is not a significant determinant of small artery structure in untreated essential hypertension. PMID- 9205415 TI - Determination of appropriate recording force for non-invasive measurement of arterial pressure pulses. AB - 1. Non-invasive recording techniques of the arterial pressure pulse will distort the arterial wall and may alter pulse wave measurements. We hypothesized that intersubject variability of these measurements would be reduced if recording forces were normalized to reflect individualized arterial occlusion forces. 2. In 10 normal male subjects (age 24 +/- 1 years), brachial, radial and finger arterial pressure pulses were recorded simultaneously using volume displacement pulse transducers (Fukuda TY-303) and a finger pressure monitoring system (Finapres, Ohmeda 2300) and were made at 2, 5 and 10-100% (10% increments) of the brachial arterial force associated with marked distortion of finger pulsations. Forces were applied at the brachial site in a randomized order while a constant 1.8 N force was applied at the radial artery site. Pressure pulses were analysed using the discrete fast Fourier transform. 3. Pulse amplitude, contour, wave velocity and relative transmission ratios remained relatively constant until the branchial artery recording force exceeded 59.9 +/- 0.3% of the largest recording force used in each subject (7.14 +/- 0.75 N). The finger pulse pressures (P < 0.0001), radial pulse amplitudes (P < 0.0001) and contours (harmonics 2-6, P < 0.003), pulse wave velocity (P < 0.021) and relative transmission ratios (harmonics 3-7, P < 0.01) then decreased with higher recording forces. 4. To avoid distortion, non-invasive recordings of arterial pressure pulse amplitude, contour, pressure wave velocity and relative transmission ratios along a peripheral arterial segment should use recording forces of less than 60% of the force associated with marked distortion of finger pulsations. PMID- 9205416 TI - Maternal serum vascular endothelial growth factor during early pregnancy. AB - 1. The objectives of the study were: (i) to investigate the serum concentrations of vascular endothelial growth factor (VEGF) in pregnant and non-pregnant women; and (ii) to study the relationship between the levels of maternal serum VEGF and the serum concentrations of human chorionic gonadotrophin (hCG) and progesterone during the first trimester. 2. Total immunoreactive VEGF was measured by competitive RIA using recombinant human VEGF165 and a polyclonal antiserum. Serum VEGF was measured in 60 non-pregnant women of childbearing age. These data were compared with serum VEGF measured in 363 women between 41 and 91 days of gestation. 3. The median serum VEGF concentration was 1.10 micrograms/l (interquartile range 0.91-1.30) in the nonpregnant women and 2.13 micrograms/l (interquartile range 1.62-2.77) in the pregnant women. Serum levels of VEGF were significantly higher among the pregnant cohort (P < 0.0001). Serum VEGF concentration was positively correlated with gestational age, increasing until ten completed weeks of pregnancy. Serum VEGF was negatively correlated with maternal height and weight, and positively correlated with serum hCG and serum progesterone (P < or = 0.0001 in all cases). Serum VEGF was lower in the pregnant women who smoked (P = 0.06). 4. Our data show a positive and highly significant correlation between maternal serum levels of VEGF and hormones reflecting placental function (hCG, progesterone). We speculate that VEGF production is increased by progesterone and hCG, and that VEGF has a positive influence on trophoblast development. VEGF may also be involved in the initiation of the maternal cardiovascular adaptation to pregnancy. PMID- 9205417 TI - Serum leptin and short-term regulation of eating in obese women. AB - 1. Leptin is generally thought to play a key role in the regulation of eating. However, its real role in human eating behaviour is still poorly known. Therefore, the role of leptin in the regulation of eating was examined in obese binge- and non-binge-eating women during exposure to food and food-related stimuli. 2. Eleven binge- and ten non-binge-eating obese women took part in the study. In addition to serum leptin, serum insulin, non-esterified fatty acids, plasma glucose, salivation, the feeling of hunger and the desire to eat were repeatedly measured during the experiment. 3. Serum leptin levels did not differ between the binge- and non-binge-eating women. Neither were leptin levels associated with the feeling of hunger or the desire to eat food, nor with the amount or composition of food eaten. During food exposure leptin levels did not change, whereas at the same time serum insulin levels increased and serum non esterified fatty acid levels decreased. The change in salivation during food exposure was inversely associated with the fasting leptin level. 4. This study indicates that serum leptin does not play a role in the regulation of eating in obese women, at least not in the short term. Furthermore, leptin levels are not different in obese binge-eating women as compared with obese non-binge-eating women. Interestingly, high fasting leptin levels may be associated with a decreased salivation response in the presence of food and food-related stimuli. PMID- 9205418 TI - Renal endothelin system in obstructive jaundice: its role in impaired renal function of bile-duct ligated rats. AB - 1. Obstructive jaundice predisposes the kidney to acute renal failure. Endothelin (ET), a potent renal vasoconstrictor and modulator of the tubular action of arginine vasopressin, has been suggested to play a pathogenetic role in acute renal failure. In the present study we therefore investigated renal function and the renal ET system in rats on day 4 after bile-duct ligation (BDL) or sham operation (SO), without (n = 7 in each group) and with treatment with bosentan, a combined ETA/ETB receptor blocker, (n = 5 in each group). 2. On day 4 after BDL, serum bilirubin had increased to 226 +/- 10 mumol/l (SEM) as compared with 6 +/- 2 mumol/l in SO rats. Endogenous creatinine clearance, an index of glomerular filtration rate, was significantly reduced to 0.7 +/0 0.1 ml min-1 g-1 of kidney weight after BDL as compared with 1.1 +/- 0.1 ml min-1 g-1 of kidney weight after SO (P < 0.05). Bosentan prevented the decrease in glomerular filtration rate (1.0 +/- 0.2 ml min-1 g-1 of kidney weight), as well as polyuria and defective concentrating ability, in BDL rats. 3. Plasma ET concentration on day 4 after surgery (28.2 +/- 1.5 pmol/l) was higher (P < 0.01) in BDL than in SO rats (12.9 +/- 1.5 pmol/l) and rose further in bosentan-treated BDL and SO rats (43.4 +/- 5.1 compared with 21.9 +/- 6.6 pmol/l). Urinary ET excretion was significantly higher in BDL rats than in SO rats (1.58 +/- 0.22 compared with 1.28 +/- 0.18 pmol 24 h-1 100 g-1 of body weight; P < 0.05). 4. ET synthesis by glomeruli isolated from BDL rats was lower [81 +/- 19 fmol h-1 (mg of protein)-1] than that from SO-rats [139 +/- 28 fmol h-1 (mg of protein)-1; P < 0.05], whereas papillary ET synthesis was higher in BDL [10 +/- 3 fmol h-1 (mg of protein)-1] than in SO rats [4 +/- 1 fmol h-1 (mg of protein)-1; P < 0.05]. 5. The results indicate that BDL is associated with increased plasma ET concentration and suppression of GFR. Enhanced renal inner medullary collecting-duct ET synthesis, which is reflected by increased urinary ET excretion, may reduce distal tubular water absorption in BDL rats. Increased circulating and renal papillary ET synthesis may thus contribute to renal dysfunction and predispose the kidney to acute renal failure in obstructive jaundice. PMID- 9205419 TI - Effect of temperature reduction on myotonia in rat skeletal muscles in vitro. AB - 1. The objective of the study was to determine the effect of temperature reduction on the response of rat skeletal muscles to myotonia-inducing agents. 2. A model myotonia was induced in the muscles in vitro, using either the chloride channel blocker anthracene-9-carboxylic acid or chloride-free Krebs solution. This model is similar in its characteristics to the myotonia which occurs in autosomal recessive generalized myotonia congenita in humans. 3. Isometric twitch contractions were recorded in the muscles in Krebs solution before and after the addition of the myotonia-inducing agent. The presence of myotonia was confirmed when the half-relaxation time of the twitch contraction after the addition of the agent was significantly greater than that before its addition. 4. Recordings were made at 37 degrees C, 30 degrees C, 25 degrees C and 15 degrees C. Myotonia developed at 37 degrees C, 30 degrees C and 25 degrees C, but not at 15 degrees C, indicating that at a temperature between 25 degrees C and 15 degrees C, anthracene-9-carboxylic acid-induced myotonia failed to develop. This supports the results obtained in humans suffering from myotonia congenita where myotonic contractions in the adductor pollicis muscle disappeared when the muscle temperature was cooled to 20 degrees C. 5. The myotonia which developed at 37 degrees C could be significantly reduced by exposure to 1 x 10(-4) mol/l ouabain or by elevation of the K+ concentration of the Krebs solution to 7.5 mmol/l. 6. Measurements made using microelectrodes showed that the conditions under which myotonia either did not develop or was significantly reduced, i.e. a temperature of 15 degrees C, exposure to 7.5 mmol/l K+ at 37 degrees C or exposure to 1 x 10( 4) mol/l ouabain at 37 degrees C were each associated with membrane depolarization. The results are discussed in terms of a possible role for depolarization in preventing/reducing the myotonic response. PMID- 9205420 TI - Hypoxia, hypocapnia and spirometry at altitude. AB - 1. Both hypoxia and hypocapnia can cause broncho-constriction in humans, and this could have a bearing on performance at high altitude or contribute to altitude sickness. We studied the relationship between spirometry, arterial oxygen saturation and end-tidal carbon dioxide (ETCO2) concentration in a group of healthy lowland adults during a stay at high altitude, and then evaluated the response to supplementary oxygen and administration of a beta 2 agonist. 2. We collected spirometric data from 51 members of the 1994 British Mount Everest Medical Expedition at sea level (barometric pressure 101.2-101.6 kPa) and at Mount Everest Base Camp in Nepal (altitude 5300 m, barometric pressure 53-54.7 kPa) using a pocket turbine spirometer. A total of 205 spirometric measurements were made on the 51 subjects during the first 6 days after arrival at Base Camp. Further measurements were made before and after inhalation of oxygen (n = 47) or a beta 2 agonist (n = 39). ETCO2 tensions were measured on the same day as spirometric measurements in 30 of these subjects. 3. In the first 6 days after arrival at 5300 m, lower oxygen saturations were associated with lower forced expiratory volume in 1 s (FEV1; P < 0.02) and forced vital capacity (FVC; P < 0.01), but not with peak expiratory flow (PEF). Administration of supplementary oxygen for 5 min increased oxygen saturation from a mean of 81%-94%, but there was no significant change in FEV1 or FVC, whilst PEF fell by 2.3% [P < 0.001; 95% confidence intervals (CI) -4 to -0.7%]. After salbutamol administration, there was no significant change in PEF, FEV1 or FVC in 35 non-asthmatic subjects. Mean ETCO2 at Everest Base Camp was 26 mmHg, and a low ETCO2 was weakly associated with a larger drop in FVC at altitude compared with sea level (r = 0.38, P < 0.05). There was no correlation between either ETCO2 or oxygen saturation and changes in FEV1 or PEF compared with sea-level values. 4. In this study, in normal subjects who were acclimatized to hypobaric hypoxia at an altitude of 5300 m, we found no evidence of hypoxic broncho-constriction. Individuals did not have lower PEF when they were more hypoxic, and neither PEF nor FEV1 were increased by either supplementary oxygen or salbutamol. FVC fell at altitude, and there was a greater fall in FVC for subjects with lower oxygen saturations and probably lower ETCO2. PMID- 9205421 TI - The quality of life of persons with chronic fatigue syndrome. AB - This descriptive study used a between-methods triangulation design to analyze the multiple dimensions of quality of life in persons with chronic fatigue syndrome (CFS). This method, which refers to the combination of both quantitative and qualitative methods in the same study, allowed the authors to obtain more comprehensive and robust data than could be obtained by either method alone. A convenience sample of 110 persons with CFS completed the quality of life index and CFS questionnaire, and a subset of 22 persons were interviewed regarding their lived experience with CFS. Overall scores on the quality of life index were significantly lower in CFS than for other chronic illness groups. Subjects reported the lowest quality of life scores in health and functioning domain. Indepth interviews provided a more complete understanding of the quality of life in CFS and further explained the low ratings that were found on the quality of life index. The findings suggest that quality of life is particularly and uniquely disrupted in CFS. PMID- 9205422 TI - The quality of life and employment in panic disorder. AB - Our purpose was to measure quality of life (QOL) and work productivity (WP) in persons with panic disorder. Eighty-four panic disorder patients with limited psychiatric comorbidity for ten U.S. outpatient mental health centers were evaluated in a cross-sectional design. Patients self-administered the Medical Outcomes Study 36-Item Short Form Health Survey (SF-36) and Work Productivity and Impairment (WPAI) questionnaire. The independent effects of psychiatric comorbidity were addressed through entry criteria, stratification, and regression analyses. QOL scores are significantly below age and sex-adjusted population norms on all SF-36 measures (p < .01). We note far greater impairment on measures of mental and emotional versus physical well-being. The unemployment rate among these patients is 25%, and only 57% are employed full-time. Those who are employed rated their WP as low. This sample of outpatients suffer marked QOL and employment impairment, which is only partially explained by the presence of psychiatric comorbidity. PMID- 9205424 TI - Somatoform disorders in Caucasian and Chinese Americans. AB - Somatization, broadly defined as the presentation of one or more medically unexplained somatic symptoms, refers both to the presentation of somatic symptoms in diagnosable psychiatric disorders such as major depression or anxiety as well as to the presentation of such symptoms in somatoform disorders. Although no comparative data exist, somatization is considered by many clinical investigators to be more common among Chinese than Caucasian patients, but it is unclear if this occurs because somatoform disorders are more prevalent among the Chinese or because Chinese patients with major depression or anxiety more often present with somatic complaints. We examined 85 consecutive Chinese American and 85 consecutive Caucasian American patients referred for psychiatric consultation and found the following: a) True somatization was significantly more common among Chinese American patients referred for psychiatric consultation; b) The somatoform symptom profiles of the two cohorts were different: Chinese American somatizers complained predominantly of cardiopulmonary and vestibular symptoms, whereas their Caucasian counterparts had symptoms that corresponded well with the categories listed in DSM-IV; c) In both cohorts of somatizers, a concurrent psychiatric disorder, most commonly major depression, was almost always present; and d) Among the Chinese American somatizers, pseudoneurological symptoms occurred most commonly in the form of abnormal sensations, whereas abnormal motor functions were more common among Caucasian Americans. Implications of the findings with respect to pathogenesis, treatment, and classification of somatization are discussed. PMID- 9205423 TI - Recent life events and suicide in personality disorders. AB - This study investigates the relationships between personality disorders, recent life events, and comorbid axis I disorders in suicide. Life events during the last week and last 3 months before suicide of 56 suicide victims with a DSM-III-R axis II personality disorder (PD) were compared with those of 56 age- and sex matched comparison suicides with evidence of no PD. These victims were from a random sample of suicides representing a total 1-year nationwide suicide population in Finland. Life events had been more common among PD victims, particularly job problems, family discord, financial trouble, unemployment, and interpersonal loss; most victims with PD had had multiple life events. Events possibly dependent on the victim's own behavior had been much more common among the PD group, especially in the final week (70% vs. 23%). Age, gender, comorbid axis I diagnosis of alcoholism or depression, or PD cluster type B or C seemed to be less important factors in terms of excess events in the PD group. Thus, PD should be assessed as an important factor when analyzing the relationship between recent life events and suicide. Our findings suggest that interpersonal and job related/financial problems may precede suicide closely among individuals with PD. PMID- 9205425 TI - Clinician ratings of the five-factor model of personality and the DSM-IV personality disorders. AB - This study explored the associations among the domains of the five-factor model (FFM) of personality (neuroticism, extraversion, openness, agreeableness, and conscientiousness) and the DSM-IV personality disorders (PDs). Clinician ratings were obtained for both the DSM-IV PDs and the FFM on a sample of 100 PD patients. The correlational data showed that the DSM PDs were most strongly associated with the FFM domains of neuroticism, extraversion, and agreeableness. Factor analysis revealed four underlying factors that provided insights into qualities shared by subgroups of the DSM-IV PDs. The domain of neuroticism was associated with the borderline, avoidant, and dependent PDs (factor 1). The paranoid, avoidant, schizoid, and schizotypal PDs were negatively associated with the domain of agreeableness (factor 2). The domain of extraversion was positively associated with the narcissistic and histrionic PDs and negatively with schizoid PD (factor 3). The FFM conscientiousness and openness domains loaded onto a single factor and were positively associated with the obsessive-compulsive PD and negatively associated with the antisocial and borderline PDs. Exploring the relationships between these two personality systems will improve our conceptualization and understanding of the DSM PDs. PMID- 9205426 TI - Traumatic brain injury in children and adolescents: psychiatric disorders in the second three months. AB - Psychiatric disorders may be common after traumatic brain injury (TBI) in children, yet there is a death of prospective studies examining this problem. Fifty children aged 6 to 14, hospitalized after TBI, were assessed soon after TBI regarding preinjury psychiatric, behavioral, adaptive, and family functioning, family psychiatric history status and injury severity. The outcome measure was the presence of a "novel" psychiatric disorder (not present before the injury) during the second 3 months after the injury. Forty-two subjects were reassessed at 6 months. Severity of injury, family psychiatric history, and family function predicted a novel psychiatric disorder. Among children suffering a mild/moderate injury, those with preinjury lifetime psychiatric disorders were no longer (as they had been in the first 3 months) at higher risk than those without such a lifetime history. Thus, there appeared to be children, identifiable through clinical assessment, at increased risk for novel psychiatric disorders after TBI. PMID- 9205427 TI - Psychological well-being and ratings of psychiatric symptoms in bereaved Israeli adolescents: differential effect of war-versus accident-related bereavement. AB - Eight hundred seventy-one Israeli adolescents, 375 boys and 496 girls, mean age 16.7 +/- 1, participated in this study. Twenty-three of them lost relatives in war and 19 in road accidents. All participants were administered the Brief Symptom Inventory (BSI), the General Well-being Scale (GWB), the Parental Bonding Instrument (PBI) and the Perceived Social Support-Family/Friend (PSS-Fa and PSS Fr) measures. War-bereaved adolescents showed significantly higher scores in psychological well-being (GWB) and significantly lower scores in reported psychiatric symptoms (BSI) than accident-bereaved adolescents. War-bereaved adolescents also had significantly better BSI and GWB scores than the general nonbereaved adolescent population. These results persisted after controlling for family socio-economic status, gender, and the degrees of closeness of the deceased relative. War-bereaved adolescents did not differ either from accident bereaved adolescents or from the nonbereaved general adolescent population in social and family support systems (PSS-Fr, PSS-Fa) and did not experience different basic parental attitudes (PBI). Results are discussed in terms of the different meanings ascribed to death in battle versus death in a road accident. PMID- 9205428 TI - Suicidal behavior in bipolar I and bipolar II disorder. PMID- 9205429 TI - Treatment outcomes for severely mentally ill patients on conditional discharge to community-based treatment. PMID- 9205430 TI - Topical penciclovir for herpes labialis. PMID- 9205431 TI - Cabergoline for hyperprolactinemia. PMID- 9205432 TI - Midodrine for orthostatic hypotension. PMID- 9205433 TI - Abdominal rescue using the vacuum extractor after entrapment of the aftercoming head. AB - BACKGROUND: Fetal replacement into the uterus for ceasarean delivery after failed vaginal delivery has been reported for both vertex and breech presentations. Although an option, this mode of delivery frequently is the last maneuver to deliver a viable infant after other methods to allow vaginal delivery have failed. We report the adjunctive use of a vacuum extractor to facilitate abdominal rescue after entrapment of the aftercoming head during an attempted vaginal breech delivery. CASE: A multiparous woman presented at term with two fetal feet bulging through the membranes at the introitus. During vaginal breech delivery, the aftercoming head became entrapped. Unsuccessful maneuvers to facilitate descent included Duhrssen incisions, the Mauriceau maneuver, placement of Piper forceps, and halothane administration. At emergency ceasarean delivery, the infant was pushed upward from below, and rapid, successful delivery of a 2530 g neonate was accomplished with assistance by a vacuum extractor. Apgar scores were 3, 6, and 7 at 1, 5, and 10 minutes, respectively. Umbilical artery blood gas revealed a pH of 7.18 and base excess of -6. A head sonogram and electroencephalogram were normal. Both mother and infant were discharged without complications. CONCLUSION: After entrapment of the aftercoming head during attempted vaginal breech delivery, use of the vacuum extractor may expedite the abdominal rescue and ceasarean. PMID- 9205434 TI - Puerperal presentation of a living abdominal pregnancy. AB - BACKGROUND: Abdominal and heterotopic pregnancies appear to be increasing in incidence. CASE: We report a case of puerperal presentation of a living heterotopic pregnancy in an African woman. The patient presented 6 days postpartum with fever and abdominal pain. The correct diagnosis of heterotopic pregnancy was not considered, and for 9 days she was treated for presumed puerperal sepsis. It was only upon abdominal x-ray that the diagnosis was made. The patient underwent laparotomy with delivery of a living male neonate weighing 2000 g. He subsequently died of respiratory failure on day 3 of life. CONCLUSION: Although still rare, the increasing incidence of abdominal pregnancies in both developed and developing countries mandates awareness of this diagnosis, particularly in pregnant or postpartum women presenting with abdominal pain. PMID- 9205435 TI - Magnetic resonance imaging in evaluation of a second-trimester ovarian twin pregnancy. AB - BACKGROUND: Ovarian pregnancy presents with abdominal pain and menstrual irregularities, and usually results in hemorrhage and hemoperitoneum in the first trimester. We describe the first case of a twin ovarian pregnancy diagnosed in the second trimester. Magnetic resonance imaging (MRI) was used in the preoperative evaluation of this patient. CASE: A woman presented at 19 weeks' gestation with abdominal pain and irregular bleeding. Her hemoglobin level was 5.9 g/dL, as compared to 10.8 g/dL in early pregnancy. Ultrasound showed a twin gestation with a mass anterior to the pregnancy, thought to be a placenta percreta or a hemorrhagic leiomyoma. An MRI was suspicious for an extrauterine pregnancy, showing the uterus displaced anteriorly by the pregnancy mass. Laparotomy revealed a hemoperitoneum and right twin ovarian pregnancy. A right salpingo-oophorectomy was performed. Pathology confirmed the diagnosis. CONCLUSION: Although ultrasound is the primary technique of imaging the pelvis during pregnancy, MRI should be considered when the ultrasound findings are limited or confusing. PMID- 9205437 TI - Orogenital contact: a cause of chorioamnionitis? AB - BACKGROUND: Fusobacterium nucleatum and Capnocytophaga species are common oral pathogens and infrequent causes of systemic infection in patients with compromised immunity or disrupted mucosal integrity. The isolation of both organisms from a clinical specimen suggests an oral source of infection. CASE: A 23-year-old black woman was admitted at 24 weeks' gestation in preterm labor. She subsequently developed signs of clinical chorioamnionitis, including fever, fetal tachycardia, and uterine tenderness. Bacteriologic studies of the amniotic fluid and subchorionic placental cultures yielded F nucleatum and Capnocytophaga species. On review of the patient's history, a temporal relation was noted between orogenital contact and the onset of clinical infection. Thorough evaluation of the patient, including dental examination, did not reveal an obvious source of infection. However, significant periodontal disease was identified in her partner. CONCLUSION: The concomitant finding of these two organisms in the patient's amniotic fluid and a history of periodontal disease in her partner suggests that chorioamnionitis may have been due to an ascending infection after orogenital contact. PMID- 9205436 TI - Use of two Greenfield caval filters to prevent recurrent pulmonary embolism in a heparin-allergic gravida. AB - BACKGROUND: Maternal mortality may be reduced by prompt diagnosis and treatment of pulmonary embolism. CASE: A 25-year-old pregnant woman required a second Greenfield filter after developing a heparin allergy and recurrent pulmonary embolism. CONCLUSION: Heparin allergy has not been reported previously in pregnancy. A Greenfield filter may be used in this circumstance. Extension of the thrombus cephalad to the filter can cause recurrent emboli, so deployment of a second caval filter may be an effective remedy. PMID- 9205438 TI - An unexpected fetal outcome following a severe maternal motor vehicle accident. AB - BACKGROUND: Maternal motor vehicle injury occurs commonly and can cause serious fetal injury. Optimum pregnancy management at the time of maternal presentation following trauma requires reliable methods of fetal assessment. In this report, we present a case in which currently accepted methods of fetal assessment initially failed to demonstrate catastrophic fetal brain injury following a maternal motor vehicle accident. CASE: A 28-year-old primigravida woman at 27 weeks' gestation was in a pedestrian motor vehicle accident, suffering a closed head injury and multiple fractures. Initial fetal assessment included cardiotocographic monitoring for 24 hours fetal ultrasound, both of which were normal, as was a biophysical profile done on the fifth day after the accident. These were repeated at intervals, but definite evidence of fetal brain injury was not seen until unilateral ventricular dilatation was documented on ultrasound at 35 weeks' gestation. Postnatal imaging showed microcephaly, hydrocephalus ex vacuo, and multiple hemispheric hypodensities, likely representing post-traumatic hemorrhages with secondary infarction. At the age of 4 years, the child is cortically blind, epileptic, and quadriparetic. CONCLUSION: This pregnancy outcome was unexpectedly poor despite the reassuring initial assessment. We caution that these methods may not provide accurate early fetal assessment, especially when fetal brain stem function is spared. PMID- 9205439 TI - Successful pregnancy following zygote intrafallopian transfer for congenital cervical hypoplasia. AB - BACKGROUND: Congenital cervical atresia and hypoplasia are rare abnormalities that generally require reconstructive or extirpative procedures to relieve outflow tract obstruction. Infertility is a common sequel, and only four previous pregnancies have been reported. In selected cases, zygote intrafallopian transfer (ZIFT) or other assisted reproductive techniques may offer alternatives for conception. CASE: A 21-year-old amenorrheic woman experienced a spontaneous gush of vaginal bleeding following an 11-year history of cyclic lower abdominal pain. Regular but prolonged and painful menses ensued. After another 8 years of primary infertility, transcervical and transfundal hysteroscopy demonstrated congenital cervical hypoplasia and a normal endometrial cavity. Conception was achieved during her third cycle of ZIFT. Delivery occurred by elective cesarean at 39 weeks for a persistent oblique fetal lie. CONCLUSION: A successful pregnancy was established following ZIFT in a woman with congenital cervical hypoplasia. The endometrial cavity was evaluated by a previously unreported technique, transfundal hysteroscopy. The use of appropriate surgical or assisted reproductive techniques in conjunction with individualized post-conception management may permit successful pregnancy and delivery in selected women with congenital cervical hypoplasia and atresia. PMID- 9205440 TI - Mixed germ cell malignancy of the ovary concurrent with pregnancy. AB - BACKGROUND: A rare malignant germ cell tumor of the ovary during pregnancy was detected by screening of maternal serum alpha-fetoprotein (MSAFP). Treatment of this uncommon tumor during pregnancy incorporated combination chemotherapy including etoposide. CASE: An 18-year-old primiparous woman undergoing antenatal genetic screening was found to have an extremely elevated MSAFP of 477.8 IU/mL, or 12.46 multiples of the median. Oophorectomy and staging laparotomy at 20.5 weeks' gestation resulted in the diagnosis of mixed germ cell tumor of the ovary, with both endodermal sinus tumor and grade 3 immature teratoma. The patient received three courses of cis-platinum, etoposide, and bleomycin. Maternal serum AFP titers had returned to normal pregnancy levels by the start of the second course. A healthy female infant was delivered at 39 weeks' gestation following induction of labor for pregnancy-induced hypertension. CONCLUSION: Elevated MSAFP levels may be a presenting sign of malignant ovarian germ cell neoplasms. This report describes both surgical and chemotherapeutic treatment of a germ cell malignancy during pregnancy, with delivery at term. To date, major fetal toxicity from chemotherapy has not been identified. PMID- 9205441 TI - Management of chemotherapy in a pregnancy complicated by a large neuroblastoma. AB - BACKGROUND: The English literature contains infrequent reports of neuroendocrine carcinoma during pregnancy. The chemotherapy for this type of malignancy can cause severe nausea and neutropenia. We used recently developed modalities to ameliorate these side effects. CASE: A 22-year-old woman, gravida 4, para 2-0-1 2, presented at 24 weeks' gestation with a complaint of massive lower-extremity edema. Computed tomography scan delineated a large retroperitoneal mass. Biopsy of a small neck mass revealed a poorly differentiated neuroblastoma. A multidisciplinary approach to therapy was undertaken. The patient received cisplatin and etoposide chemotherapy. Complications of the first course included severe neutropenia and nausea with vomiting. Filgrastim and ondansetron were used to treat these complications. She delivered an 1825-g healthy male by cesarean for fetal distress at 35 weeks. No anomalies were noted at birth. Neonatal hematologic indices were normal CONCLUSION: A multidisciplinary approach to rare malignancies is warranted in pregnancy. Filgrastim and ondansetron are effective agents in the treatment of chemotherapy-associated complications. Their use in pregnancy warrants further investigations. PMID- 9205442 TI - Pulmonary artery sarcoma in a pregnant woman: report of a case. AB - BACKGROUND: We report the case of a 23-year-old pregnant woman with hemoptysis, cor pulmonale, and pulmonary artery sarcoma. The physiologic changes of pregnancy may have unmasked the pulmonary lesion. CASE: A 23-year-old woman presented at 28 weeks' gestation with acute onset of hemoptysis and dyspnea. A hilar mass was noted and a pulmonary embolus was diagnosed. Biopsy of the hilar mass was nondiagnostic. Emergency cesarean delivery was performed because of rapid clinical deterioration and an acute loss of fetal heart tones. Both mother and infant died. Autopsy of the mother demonstrated a large pulmonary artery sarcoma with metastases to both lungs and terminal bacterial bronchopneumonia. CONCLUSION: Hemodynamic changes of pregnancy may have unmasked the pulmonary lesion in this case. Pulmonary artery sarcoma is an extremely rare tumor. PMID- 9205444 TI - Non-oral pyogenic granuloma in pregnancy: a report of two cases. AB - BACKGROUND: Pyogenic granulomas are benign vascular lesions of the skin or mucous membranes. Oral lesions are believed to occur in up to 2% of pregnancies. To the best of our knowledge, non-oral lesions in pregnancy have not been reported in the obstetric literature. CASES: We report two cases of non-oral pyogenic granuloma in pregnancy. The first, involving a finger lesion in a woman with triplets, demonstrated rapid growth and recurred after surgical excision. The second was an inguinal crease lesion, which was excised successfully after becoming symptomatic. CONCLUSION: Clinical experience suggests that the prevalence of pyogenic granulomas in pregnancy is not as high as has been reported in the literature. The relation of non-oral lesions to pregnancy is unknown. PMID- 9205443 TI - Crigler-Najjar disease in pregnancy. AB - BACKGROUND: Crigler-Najjar disease, a rare cause of maternal unconjugated hyperbilirubinemia in pregnancy, poses no threat to the mother, and the elevated bilirubin levels do not seem harmful to the fetus. However, the disease is expressed in two forms, one of which is fatal. CASE: The maternal total bilirubin (mostly unconjugated) was 8.5 mg/dL in the first trimester, fell to 5.0 mg/dL in the second, and rose again to 8.8 mg/dL at term. The infant was jaundiced at birth, with umbilical cord total bilirubin at 7.6 mg/dL. The jaundice resolved without treatment, and no sequelae of hyperbilirubinemia were present. CONCLUSION: Crigler-Najjar disease type II seems to pose no unique maternal risk during pregnancy. The fetus seems to be resistant to elevated maternal unconjugated bilirubin, but the neonate may required therapy for hyperbilirubinemia. PMID- 9205445 TI - Right atrial thrombus as a complication of total parenteral nutrition in pregnancy. AB - BACKGROUND: Right atrial thrombus, a rare but potentially fatal complication of central venous catheter use in total parenteral nutrition, has not been reported during pregnancy. CASE: A pregnant woman with persistent hyperemesis gravidarum developed a right atrial thrombus related to central venous catheter use for total parenteral nutrition. CONCLUSION: Right atrial thrombus can be treated successfully with heparin, leading to its resolution and a normal pregnancy outcome. PMID- 9205447 TI - Intrapartum, atraumatic, non-asphyxial intracranial hemorrhage in a full-term infant. AB - BACKGROUND: Intracranial hemorrhage in a full-term infant is uncommon, is usually subarachnoid in type, and is usually associated with operative vaginal delivery or asphyxia. CASE: A 15-year-old primigravid woman at 37 weeks' gestation developed a prolonged second stage of labor associated with persistent occiput posterior position. With the onset of pushing, baseline fetal heart rate (FHR) decreased and variability increased. Thirty minutes before vaginal delivery, sudden fetal tachycardia (up to 210 beats per minute) was observed, with absent variability and minimal decelerations. At birth, the infant was apneic and hypotonic, but lacked biochemical evidence of acidemia or asphyxia; seizures developed in the early neonatal period. Subarachnoid hemorrhage was demonstrated by computed tomography of the head. CONCLUSION: The occiput posterior position, marked molding, and prolonged labor with compulsive pushing may be associated with an increased risk of adverse outcome, even unrelated to the details of delivery. The change in FHR pattern, to a lowered baseline rate and increased variability, suggests increased intracranial pressure. The sudden change to fetal tachycardia with absent variability before delivery suggests intracranial hemorrhage or injury. PMID- 9205446 TI - Pregnancy complicated by liver dysfunction: possible pathogenesis of vasospasm. AB - BACKGROUND: Acute fatty liver of pregnancy, the syndrome of hemolysis, elevated liver enzymes, and low platelet count (HELLP), and severe preeclampsia form a disease spectrum. We report a case that showed a vasospastic phenomenon supported angiographically and hematologically. CASE: A 31-year-old Japanese woman presented at 37 weeks gestation with a 1-week history of nausea, vomiting and general fatigue. She underwent cesarean delivery for fetal distress. Liver dysfunction and disseminated intravascular coagulopathy were detected. The celiac angiogram showed vascular narrowing and irregularity of the vascular wall. Serum endothelin and the ratio of thromboxane B2 to 6-keto-prostaglandin F1 alpha were increased at the same time. The patient experienced rapid resolution of symptoms and laboratory abnormalities in the immediate postoperative period. CONCLUSION: A pregnancy complicated by liver dysfunction showed a vasospastic phenomenon, which may suggest the presence of a vasospastic syndrome. PMID- 9205448 TI - Intrapartum fetal surveillance of congenital heart block with pulse oximetry. AB - BACKGROUND: In cases of fetal congenital heart block, the fetal heart rate (FHR) pattern is uninterpretable, often leading to an operative delivery. Reflectance pulse oximetry, a new technique that continuously measures the fetal arterial oxygen saturation (SaO2) during labor, is potentially useful in intrapartum monitoring of fetuses with this condition. CASES: Two fetuses with congenital heart block were monitored with reflectance pulse oximetry and fetal scalp blood sampling. The first patient delivered spontaneously. Adequate signal quality was achieved during 73% of the study time. Mean +/- standard deviation (SD) SaO2 was 53 +/- 14%. Fetal outcome was good. The second patient was delivered by cesarean because of arrest of labor. Oxygen saturation values were obtained during 89% of the study time. The mean SaO2 was 42 +/- 13%. There was a period of 8 minutes with SaO2 values below 20%. Capillary blood pH dropped from 7.33 to 7.25; SaO2 values then returned to levels above 30% and the capillary blood pH normalized. The neonate was born in good condition. CONCLUSION: In fetal congenital heart block, adequate surveillance with FHR monitoring during labor is not possible; therefore, continuous information on fetal oxygenation may be valuable in assessing the fetal condition and may prevent unnecessary obstetric interventions. PMID- 9205449 TI - Ectopic atrial tachycardia in utero. AB - BACKGROUND: The antenatal diagnosis of fetal arrhythmias helps direct medical management, which may include intrauterine therapy. Ectopic atrial tachycardia is an unusual arrhythmia in children, and we know of no previous reports of antenatal diagnosis of this particular arrhythmia. CASE: The diagnosis of fetal ectopic atrial tachycardia was suggested by the monitor tracing during labor and was subsequently confirmed by the postpartum behavior of the arrhythmia and its electrocardiographic characteristics. CONCLUSION: Ectopic atrial tachycardia, although uncommon, should be considered in the differential diagnosis of fetal tachycardias. The fetal monitor tracing may be useful in making this diagnosis antenatally, which may help to direct management both before and after birth. PMID- 9205450 TI - Prenatal diagnosis of an unusual nuchal cord complication in monoamniotic twins. AB - BACKGROUND: Monoamniotic twin pregnancies are frequently associated with cord entanglement, but such entanglement rarely involves the co-twin's trunk, extremities, or neck. CASE: We report a set of monoamniotic twins in which color Doppler imaging revealed that the cord of twin B was wrapped around the neck of its dead co-twin. This knowledge allowed us to avoid clamping and dividing twin A's nuchal cord during vaginal delivery, preventing asphyxia of twin B. This is the fifth reported incidence of this particular monoamniotic complication and the first to be diagnosed prenatally. CONCLUSION: Color Doppler imaging facilitates the diagnosis of rare cord complications in monoamniotic twin pregnancies. PMID- 9205451 TI - Transient hydrops fetalis associated with intrauterine cytomegalovirus infection: prenatal diagnosis. AB - BACKGROUND: Intrauterine cytomegalovirus infection is usually unrecognized during pregnancy. However, in some cases, ultrasound abnormalities can be observed in association with cytomegalovirus infection. CASE: The prenatal diagnosis of cytomegalovirus infection in a fetus with transient hydrops is reported. Fetal ascites was first recognized by routine ultrasound examination at 20 weeks' gestation. Hydrops fetalis was obvious at 23 weeks and completely resolved 1 week later. Cytomegalovirus was detected from amniotic fluid samples by centrifugal culture and direct immunofluorescent examination. The diagnosis of maternal primary infection could be established retrospectively by demonstrating immunoglobulin (Ig) G and IgM seroconversion on sequential sera. The pregnancy was electively terminated. Autopsy findings were consistent with fetal disseminated infection. CONCLUSION: Transient hydrops fetalis in association with intrauterine cytomegalovirus infection is infrequent. The resolution of hydrops fetalis could be explained by hepatic dysfunction of limited duration. Amniotic fluid culture is a reliable approach for diagnosing intrauterine cytomegalovirus infection, but does not predict the severity of the disease or the outcome of the pregnancy. The long-term clinical significance of intrauterine cytomegalovirus infection has to be established. PMID- 9205452 TI - Fetal compromise associated with extreme fetal bile acidemia and maternal primary sclerosing cholangitis. AB - BACKGROUND: Primary sclerosing cholangitis is a rare form of progressive biliary inflammation and fibrosis of unknown etiology that ultimately destroys the liver, leading to cirrhosis, liver failure, and death. Only one prior case has been reported in pregnancy. CASE: Our patient had the diagnosis of primary sclerosing cholangitis made 2 years before conception. Her course was remarkable for retroplacental hemorrhage at 14-16 weeks and preplacental hemorrhage at 28-34 weeks, with fetal growth retardation, spontaneous premature rupture of the membranes, meconium-stained amniotic fluid, fetal bradycardia, and peripartal exacerbations of her disease. Most uniquely, however, in our case the fetal compromise was associated with an extreme elevation of the bile acid level to greater than 2000 mg/dL in the fetal circulation. CONCLUSION: The extreme elevation of fetal bile acidemia at levels greater than 40 times normal in our case may well represent the pathophysiologic link between aberrations of maternal bile acid metabolism during pregnancy and fetal compromise. PMID- 9205453 TI - Uncommon location of persistent ectopic pregnancy following laparoscopic surgery. AB - BACKGROUND: Laparoscopic surgery has become an important tool in the treatment of tubal pregnancy. Skillful operative technique should prevent tissue spread and thus avoid persistence of ectopic trophoblastic cells. CASE: A laparotomy with complete salpingectomy was performed in a patient with rising serum hCG levels after a previous laparoscopic partial salpingectomy for ampullary tubal pregnancy. The only residual trophoblastic tissue found was an implant in the abdominal wall at the site of auxiliary puncture. CONCLUSION: Extra-abdominal dispersion of active trophoblastic cells may lead to increasing hCG levels, mimicking persistent tubal pregnancy. PMID- 9205454 TI - Ovarian torsion: diagnosis by color Doppler ultrasonography. AB - BACKGROUND: Ovarian torsion, although a rare gynecologic emergency, is a threat to women of all ages. Traditionally, ultrasonography and laparoscopy facilitated early diagnosis and treatment of this condition. This case report highlights the usefulness of color Doppler ultrasonography in the diagnosis of ovarian torsion. CASE: A 25-year-old patient who conceived after ovulation induction was treated conservatively for mild ovarian hyperstimulation and threatened abortion. She subsequently underwent selective first-trimester multifetal reduction and was admitted and treated for suspected pelvic infection. On the 17th post-procedure morning, ovarian torsion was diagnosed using color Doppler ultrasonography. CONCLUSION: Because gynecologists face serious management dilemmas when confronted with ovarian torsion, this technique of using color Doppler ultrasonography should provide a highly specific finding in complete ovarian torsion, aiding the clinician in prompt diagnosis and treatment. PMID- 9205455 TI - Duplicate cervix and vagina associated with infertility, endometriosis, and chronic pelvic pain. AB - BACKGROUND: Mullerian anomalies are associated with several gynecologic complications including endometriosis, infertility, and pelvic pain. CASE: A woman with duplicate cervix and a non-communicating longitudinal vaginal septum, but no other uterine anomalies, presented with pelvic pain, secondary infertility, and a long history of endometriosis. She was treated with operative laparoscopy and excision of the vaginal septum. CONCLUSION: A thorough evaluation, including history, physical examination, and appropriate imaging techniques (hysterosalpingography and magnetic resonance imaging) facilitates accurate diagnosis of anatomical defects and any associated disease in cases of unusual mullerian anomalies. An accurate preoperative diagnosis allows a planned, efficient surgical approach. PMID- 9205456 TI - Wide-band transabdominal cerclage for a foreshortened, incompetent cervix. AB - BACKGROUND: The role of cervical cerclage in the prevention of fetal wastage due to cervical incompetence is well established. The transvaginal approach and, failing that, the transabdominal approach, provide sufficient treatment in most cases. However, the traditional techniques require adequate cervical length for placement and maintenance of the suture. CASE: We report a new technique used for a patient with a markedly foreshortened cervix and a history of multiple second trimester pregnancy losses despite placement of McDonald cerclages. To improve the performance of the cervix, we included the lower portion of the uterus in a 3 cm-wide Prolene mesh cerclage. During the patient's subsequent pregnancy, the mesh band funneled the lower uterine segment, creating a functionally longer cervix. The patient successfully carried the pregnancy to term and was delivered by cesarean. CONCLUSION: This variation on the transabdominal approach is useful in the management of patients with cervical incompetence who demonstrate a foreshortened cervix incapable of maintaining a traditional cervical suture. PMID- 9205457 TI - Ruptured pelvic appendix diagnosed by transvaginal sonography. AB - BACKGROUND: Appendicitis in a pelvic appendix can be difficult to diagnose. Transvaginal sonography may help to visualize an inflamed pelvic appendix. CASE: A 31-year-old woman presented to the hospital with symptoms suggestive of pelvic inflammatory disease. Transabdominal ultrasound, useful in the diagnosis of appendicitis, showed a mass between the uterus and the right ovary. Transvaginal ultrasound clarified the finding as a bulbous fluid-filled structure extending into the cul-de-sac. The structure was diagnosed as an inflamed appendix. Laparoscopic appendectomy was performed, and the patient had an uneventful recovery. CONCLUSION: To our knowledge, this is the first case of appendicitis diagnosed with transvaginal sonography. Transvaginal sonography can delineate the features of an inflamed pelvic appendix and help to narrow the diagnostic possibilities in symptomatic women of childbearing age. PMID- 9205458 TI - Hemangioma of the uterus associated with hereditary hemorrhagic telangiectasia. AB - BACKGROUND: Hemangiomas of the uterus are rare. Involvement of the uterus with hereditary hemorrhagic telangiectasia causing menorrhagia is also rare. To our knowledge, only one case of combined uterine hemangioma and hereditary hemorrhagic telangiectasia has ever been reported. CASE: A 34-year-old woman was to undergo hysterectomy for menorrhagia unresponsive to treatment. Before surgery, she was found to have typical telangiectases associated with hereditary hemorrhagic telangiectasia. The fundus of the uterus contained a hemangioma extending from the serosa to the endometrium. CONCLUSION: Vascular malformation have been found in various organs in individuals with hereditary hemorrhagic telangiectasia. Although involvement of the uterus in hereditary hemorrhagic telangiectasia is uncommon, telangiectasia should be considered in any patient with menorrhagia resistant to treatment. PMID- 9205459 TI - Malassezia furfur folliculitis of the vulva: olive oil solves the mystery. AB - BACKGROUND: In treating women with chronic fungal infections, it is important to know which organism is responsible for the infection. In the past, organisms thought to cause vaginitis and vulvitis could all be cultured on modified Sabouraud agar. CASE: We describe a case of a woman whose chronic fungal vulvar folliculitis masqueraded as squamous epithelial hyperplasia. The 46-year-old woman, taking immunosuppressive therapy for rheumatoid arthritis, was referred with an 8-month history of vulvar vesicles, itching, and burning. Her examination revealed a vulvar folliculitis. When fungal cultures were initially negative, a vulvar biopsy revealed a squamous epithelial hyperplasia. However, a fungal culture covered with sterile olive oil eventually grew Malassezia furfur, a yeast with peculiar growth requirements. She was cured with a 2-week course of fluconazole. CONCLUSION: Malassezia furfur, an organism rarely described in the vaginitis literature, can cause vulvar folliculitis in a patient on immunosuppressive therapy. PMID- 9205460 TI - Idiopathic CD4+ T-lymphocytopenia and recurrent vulvar intraepithelial neoplasia. AB - BACKGROUND: CD4+ T-lymphocytopenia immunodeficiency without human immunodeficiency virus (HIV) infection has been reported recently. The association between immunodeficiency and anogenital neoplasia secondary to human papillomavirus infections is well documented. CASE: A woman with recurrent vulvar intraepithelial neoplasia (VIN) had idiopathic CD4+ T-lymphocytopenia without HIV infection. CONCLUSION: Human papillomavirus-related VIN may be associated with idiopathic CD4+ T-lymphocytopenia. PMID- 9205461 TI - Distribution of intraperitoneal chemotherapy into the pleural cavity. AB - BACKGROUND: Intraperitoneal chemotherapy is an established treatment for abdominal and pelvic malignancies. Several catheter-related complications have been reported. We report a case of abnormal distribution of intraperitoneal chemotherapy into the pleural cavity. CASE: A patient receiving intraperitoneal cisplatin developed shortness of breath. A pleural effusion was diagnosed and was evacuated by thoracentesis. Abnormal distribution of instilled fluid was responsible for her distress. CONCLUSION: Communications exist between the peritoneal and pleural cavities. The use of intraperitoneal chemotherapy or radioactive material may lead to respiratory complications if abnormal distribution occurs. PMID- 9205462 TI - Granulosa cell tumor of the ovary associated with antecedent tamoxifen use. AB - BACKGROUND: Increased attention has been focused recently on the estrogenic effects of tamoxifen. Review of the literature reveals an association between tamoxifen use and gynecologic tumors. CASE: A 52-year-old postmenopausal woman was treated with tamoxifen for stage II estrogen receptor-positive breast carcinoma. Her aspartate transaminase and alanine transaminase levels increase markedly after 6 months of tamoxifen use. After an additional 17 months of elevated serum transaminases, the patient was found to have a stage Ic granulosa cell tumor of the ovary. CONCLUSION: Patients with tamoxifen-induced liver dysfunction may be at increased risk for granulosa cell tumors because of alterations in tamoxifen metabolism. PMID- 9205463 TI - Growing teratoma syndrome after chemotherapy for germ cell tumors of the ovary. AB - BACKGROUND: The growing teratoma syndrome has been described with regard to gonadal and extragonadal germ cell neoplasms in males, but few cases have been reported in the female population. In this condition, masses that enlarge during or after chemotherapy are found to contain mature teratoma without malignant elements. CASES: Three patients had either persistent or growing masses despite chemotherapy for germ cell malignancies of the ovary. All cases fit the description of the growing teratoma syndrome. The patients were aged 20-22 years. All three patients had immature teratomas before chemotherapy. The stages of disease ranged from Ia to IIIc. All patients had normal tumor markers while their masses showed growth or persistence. All were free of disease 6-31 months after diagnosis. CONCLUSION: Growth or persistence of a tumor after chemotherapy for malignant teratoma does not necessarily imply progression of malignancy, especially if tumor markers are normal. However, these masses should be resected because they may cause obstruction, compression, or displacement of adjacent organs, or undergo sarcomatous degeneration. PMID- 9205464 TI - Giant uterine fibromyoma producing secondary polycythemia. AB - BACKGROUND: Although the association between large uterine fibromyomas and secondary polycythemia has been described previously, the mechanism has not been elucidated definitively. Investigators have measured erythropoietin levels in fibromyomas to determine whether these tumors are causing the polycythemia by erythropoietin overproduction; however, these studies were performed before the availability of recombinant erythropoietin assays. CASE: A 59-year-old woman presented with a 3-year history of polycythemia. Pelvic examination revealed a large lower abdominal mass. Laboratory evaluation revealed a hemoglobin of 20.8 g/dL, red blood cell mass of 3300 mL, oxygen pressure of 58 mmHg with an oxygen saturation of 89%, and erythropoietin level of 18 mU/mL. Cardiac echocardiogram showed no evidence of shunt. Computed tomography scan of the abdomen showed a large mass arising in the pelvis and compressing both ureters. The patient was treated surgically with a total abdominal hysterectomy. Pathology confirmed a uterine leiomyoma weighing 2320 g. Two months post-surgery, the patient was asymptomatic with a hemoglobin of 13.9 g/dL and erythropoietin level less than 4.0 mU/mL. CONCLUSION: This case provides evidence for three of the postulated mechanisms by which uterine fibromyomas may cause polycythemia. First, the patient was hypoxic, suggesting shunting within the tumor. Second, the leiomyoma was compressing the ureters, so the kidneys may have been inappropriately producing excess erythropoietin. Third, the tumor itself may have been producing the erythropoietin. In any case, the erythropoietin level in this patient was inappropriately high, providing useful evidence that her polycythemia was secondary to her fibromyoma. PMID- 9205465 TI - Papillary serous cystadenocarcinoma arising in benign glandular inclusion cysts in pelvic and inguinal lymph nodes. AB - BACKGROUND: Benign glandular inclusion cysts occurring within lymph nodes have been well described in the literature. However, the malignant potential of these glands is unknown. One previous case report described an adenoacanthoma arising within one of these glands. CASE: A 65-year-old woman was previously diagnosed with papillary serous cystadenocarcinoma in the inguinal and pelvic lymph nodes. She had no tumor involving the ovaries or peritoneal surfaces at the time of initial diagnosis. She presented to us 9 years later with a recurrence of this tumor in the obturator fossa and along the vaginal sidewall. Treatment consisted of surgery, radiation, and chemotherapy. CONCLUSION: Although rare, mullerian tumors can occur in the lymph nodes without simultaneous ovarian or peritoneal involvement, and most likely arise de novo within lymph node inclusion cysts. PMID- 9205466 TI - Iatrogenic endometrial megapolyps in women with breast carcinoma. AB - BACKGROUND: Tamoxifen, a widely used drug in adjuvant therapy of breast carcinoma, is now being tested for its effectiveness in chemoprevention. Although its side effects are few, tamoxifen increases the incidence of proliferative lesions of the endometrium, which theoretically should be preventable with progestational agents. CASES: Two postmenopausal women treated with tamoxifen and progestational agents for breast carcinoma developed uterine enlargement and intermittent spotting. Hysterectomy revealed benign endometrial megapolyps with marked stromal decidualization and edema. CONCLUSIONS: The value of multihormonal therapy in breast carcinoma is not established, and the addition of progestogens to tamoxifen may not reduce of developing endometrial lesions, including carcinoma. In both our cases, such a regimen did not prevent the occurrence of endometrial polyps which, although histologically benign, were usually large and thought clinically to be malignant. Periodic gynecologic assessment should be part of the follow-up of all women on long-term tamoxifen therapy. PMID- 9205467 TI - Anal incontinence and the obstetrician-gynecologist. AB - OBJECTIVE: To gather, synthesize, and present useful scientific information concerning the anal continence mechanism that will aid obstetrician-gynecologists in managing vaginal birth and evaluating women with anal incontinence not caused by disruption of the external anal sphincter. DATA SOURCES: Sources included a Medline search and reference lists of relevant articles and standard textbooks. METHODS OF STUDY SELECTION: Articles were identified that contained scientific data on the pathophysiology of anal incontinence, the influence of vaginal delivery on the continence mechanism, and therapeutic measures. Only those presenting original research results were included. Studies concerned exclusively with surgical management of the ruptured perineum were excluded. DATA EXTRACTION AND SYNTHESIS: All articles were reviewed and the physiologic data summarized. These findings were grouped by their relevance to each anatomical or physiologic issue involving anal incontinence and by whether they considered the issue of injury at the time of vaginal delivery. The data were then assembled into a functionally oriented overview of the continence mechanism. The subject of injury at the time of vaginal delivery was considered separately against a background of continence pathophysiology. CONCLUSION: Vaginal delivery may initiate damage to the continence mechanism by direct injury to the pelvic floor muscles, damage to their motor innervation, or both. Additional denervation may occur with aging, resulting in a functional disability many years after the initial trauma. These factors should be kept in mind when conducting vaginal birth and planning therapy for anal incontinence. PMID- 9205468 TI - [Prospective study of pathogenic agents isolated from feces of patients with HIV infections]. AB - OBJECTIVE: Determine the frequency of enteropathogenic agents isolated in diarrheic feces of patients with HIV infection and to compare findings with a control group (HIV + without diarrhea) in order to identify risk factors. PATIENTS AND METHODS: All HIV seropositive inpatients and outpatients seropositive for HIV, with or without diarrhea, seen between 1 November 1994 and 30 April 1995 were included. Samples of feces were obtained for culture, virology examination, parasite examination and search for Clostridium difficile. The same samples were obtained in case of diarrhea during the course of hospitalization. RESULTS: There were 113 samples. Analyses demonstrated a pathogenic agent in 73.6% of the samples in patients with diarrhea and in 31.6% of those without diarrhea. Clostridium difficile and parasites were the most frequently identified agents. An infectious agent was identified in one-fourth of the patients without clinical signs of diarrhea, and in one-fourth of those with diarrhea no pathogen could be demonstrated. No factor of risk for finding a particular microorganism in feces of patients with diarrhea could be identified. DISCUSSION: The exact pathogenic roles of Pseudomonas aeuriginosa, yeast, and adenovirus remain to be determined. It is hypothesized that the HIV has a direct effect on the host digestive tract. PMID- 9205469 TI - [Thyroid nodules: assay of serum thyrotropin and cytopuncture versus scintigraphy as first-line tests. Prospective study with 150 cases]. AB - OBJECTIVE: Fine-needle aspiration (FNA) is now considered as the first-line investigation for the diagnosis of thyroid nodules. We searched for a more accurate and cost-effective methodology as this technique fails to recognized hot nodules, frequent in certain countries. PATIENTS AND METHODS: A prospective study was conducted in 150 patients to compare two diagnostic procedures: scintigraphy first combined with FNA in case of cold nodules versus TSH measurement plus FNA when TSH measurement plus FNA when TSH was not depressed. The results were subjected to cost/benefit analysis. RESULTS: Cystic nodules were found in 28 cases (including 3 hyperfunctionning nodules, with 5 suspicious smears (1 carcinoma). FNA was non-diagnostic in 26 patients; 12 were operated on (1 carcinoma), 14 had further FNA (5 suspicious, 9 benign). Altogether 56 nodules were removed, for toxic adenoma (n = 5), for suspicious (n = 21) or malignant (n = 12) smear, or on personal (n = 18) demand; 16 carcinomas were found (2 medullary, 13 capillary, 1 follicular carcinomas). With scintigraphy first, the cost was 787 French francs (FF) per patient. With TSH measurement and FNA, the cost was 554 FF per patient. In both cases, the same number of carcinomas were removed, and all the hot nodules (11 including 5 toxic adenomas) were detected. CONCLUSION: Serum TSH measurement, with scintigraphy if TSH is low, and FNA in all the other cases, is accurate and more cost-effective than scintigraphy as a first-line investigation for the diagnosis of thyroid nodule. PMID- 9205470 TI - [Cerebrovascular complication in non-bacterial thrombotic endocarditis. Value of cardiac transesophageal ultrasonography]. AB - BACKGROUND: Non-bacterial thrombotic endocarditis in patients with cancer can lead to ischemic stroke. Endocardial vegetations are usually small and may be missed at transthoracic echocardiography. CASE REPORT: Disseminated intravascular coagulation developed in a woman with ischemic stroke. Transthoracic echocardiography was normal. Four days later, transesophageal echocardiography revealed a large mitral vegetation suggesting non-bacterial thrombotic endocarditis. The diagnosis was confirmed at pathology which reported carcinoma of the colon. DISCUSSION: Transthoracic echocardiography is rarely contributed to the diagnosis of thrombotic endocarditis. In our patient transesophageal echocardiography grave the diagnosis before death instead of retrospectively at autopsy as usually occurs, demonstrating the value of transesophageal echocardiography for cancer patients who develop ischemic stroke. PMID- 9205471 TI - [Aneurysmal dilatation of the left auricle of heart]. PMID- 9205472 TI - [Anaphylactoid manifestations after ingestion of tuna: think about scombroid intoxication]. PMID- 9205473 TI - [Lichen planus after hepatitis B vaccination. 3 new cases]. PMID- 9205474 TI - [Pulmonary embolism in a HIV-positive patient using estrogens by the percutaneous route]. PMID- 9205475 TI - [Biological diagnosis of whooping cough: contribution of gene amplification]. AB - An upsurge in pertussis infections, despite mandatory vaccination in France since 1966, has occurred again in developed countries due to progressive loss of vaccinal immunity and wider circulation of the causal bacterium, Bordetella pertussis. Unfortunately, the classical culture method is insufficiently sensitive and serology can only confirm diagnosis retrospectively. New techniques are needed for rapid diagnosis, and subsequent treatment and preventive measures. One new method, gene amplification using polymerase chain reaction (PCR), has been particularly useful in detecting Bordetella pertussis. PCR is highly specific and more sensitive than culture. It is thus quite useful in case of atypical clinical presentations and in previously treated or vaccinated patients. Less restrictions on sample transportation and preservation make PCR a technique which general practitioners can use for rapid easy diagnosis of pertussis. PMID- 9205476 TI - [Medicalized interhospital transportations and immediate outcome of patients according to the duration of hospitalization after transportation (more or less than 24 hours)]. PMID- 9205477 TI - [Acquired Willebrand's syndrome in a patient with gastric MALT lymphoma]. PMID- 9205478 TI - [Leptin: a genetic solution to obesity?]. AB - LEPTINE: Produced by adipose tissue, leptine is a regulatory hormone controlling body fat mass. GENETICS: The human gene coding for leptine was first cloned in 1984. Its receptor, a member of the class 1 cytokine receptor family, has also been identified. VARIABLE SERUM LEVELS: Plasma levels of leptine in normal-weight subjects are in the 5ng/ml range and reach 50 ng/ml in obese subjects. Weight gain leads to higher blood levels and weight loss to lower levels. The effect of leptine is to approach a weight equilibrium. Leptine level is correlated with energy balance. MODE OF ACTION: Leptine acts on hypothalamic centers controlling satiety. One of the essential mediators is neuropeptide Y. Resistance to leptine has been evidenced in human obesity. Resistance can occur as several levels and would be one of the explanations for massive obesity involving genetic factors. Leptine may also play a role in certain types of infertility. PMID- 9205479 TI - [Treatment of rectovaginal endometriosis]. AB - FREQUENT: The most frequent deep localization, endometriosis of the rectovaginal septum may penetrate into the vagina or rectum. MEDICAL TREATMENT: Hormone therapy suppresses menstruation if a short-term regimen is given, but the cost of long-term therapy with LHRH analogues and estrogen substitution may be high. SURGERY: Definitive treatment is difficult to achieve, but surgery may be required. The risks of surgery for this benign disease must be weighed against the expected benefit, particularly in drug-resistant cases where pain is severe or rectal symptoms predominant. ASYMPTOMATIC DISEASE: No specific treatment is required in asymptomatic patients or when unpainful disease is discovered at infertility explorations. PMID- 9205480 TI - [New gram-positive opportunistic bacteria: pathogenic role and treatment]. AB - NOVEL BACTERIA: The appearance of novel bacterial species among Gram positive microorganisms is mainly related to progress in bacterial taxonomy justifying such nomen species, i.e. C. jeikeium, C. urealyticum or R. equi. Another feature of such emergence of "Novel" bacteria is related to the rise in the number of clinical observations mediated by some well-known species described elsewhere such as in food bacteriology. PREDISPOSING FACTORS: Among Gram positive microorganisms, emergence for some species and renaissance or rebirth for others is mainly explained by natural resistance to widely used antibiotics including beta-lactams such as third generation cephalosporins, aminoglycosides, or more recently glycopeptides and the combined effect of several predisposing factors (hospitalized patients, underlying disease or prematurity, interruption of the normal integumentary defense via intravascular catheters). Clinical features depend on the bacterial species. Bacteriological diagnosis is easily obtained in terms of isolation and identification. Nevertheless, as for any opportunistic pathogen, a distinction must be made between colonization and infection. Finally successful treatment regimens depend on the bacterial species and may include surgery. PMID- 9205481 TI - [Cardiovascular manifestations in systemic scleroderma]. AB - OBJECTIVES: Patients with systemic scleroderma often have latent heart disease which could play an important role in morbidity and mortality. We therefore conducted a prospective study of cardiovascular manifestations in patients with systemic scleroderma. PATIENTS AND METHODS: A prospective cross-sectional study included 29 patients with systemic scleroderma who underwent a complete cardiovascular work-up including physical examination, electrocardiogram, chest x ray and Doppler-echocardiogram from July 1993 to February 1996. RESULTS: Hypertension was observed in 6 patients (20.7%) and was positively correlated with age (p = 0.007). Raynaud syndrome was also found in 6 patients (20.6%). Heart disease was observed in 14 patients (48.3%) and was positively correlated with age and lack of treatment for scleroderma (p = 0.008). Myocardial disease was the most frequent (11 patients, 37.9%), followed by pericardial disease and valve disease (4 cases each, 13.8%). Rhythm and conduction disorders were found in 2 (6.9%) and 8 (27.6%) of the patients. CONCLUSION: Cardiovascular manifestations are frequent but often latent in patients with systemic scleroderma. This finding emphasizes the importance of routine cardiovascular work-up in all patients with scleroderma. PMID- 9205482 TI - [Esophageal pH-metry. Evaluation of prescriptions at the Robert-Debre Hospital (Paris)]. AB - OBJECTIVES: To record orders for esophageal pH-metry at the Robert-Debre hospital in Paris and to assess the quality and pertinence on the order. METHODS: A prospective survey of all children who underwent pH-metry during the month of February 1996 was conducted. There were 11 children included in the study. RESULTS: Half of the pH-metries were performed in inpatients and half in outpatients including 18% scheduled directly by the exploration unit. The order did not provide any reason for the exploration in 15% of the cases (42% of those scheduled by the unit). When a reason was explained, the order described the clinical context in 86% of the cases but did not give any information on ongoing treatment in 40%. Clinical indications mentioned respiratory or ear-nose-throat disease in 70% of the cases and regurgitations or vomiting in 42%. Pathological reflux was diagnosed in one-third of the explorations. An anti-reflux treatment was prescribed for half of the patients. In reference to the official prescription guidelines in France (Reference Medicale Opposable, RMO), 72% of the orders were pertinent, 10% were questionable, 10% did not comply to official guidelines, and 8% could not be classified due to lack of information. CONCLUSION: These findings point out that the official guidelines may be of questionable value in the hospital setting. PMID- 9205483 TI - [Duodenal tuberculosis: a difficult diagnosis]. AB - BACKGROUND: The polymorphous clinical presentation of tuberculosis located in the duodenum may mislead diagnosis. CASE REPORT: A 69-year-old man had duodenal stenosis associated with calculous common bile duct obstruction. Crohn's disease was initially diagnosed and the patient was treated with corticosteroids. Two months later, the diagnosis was rectified when pulmonary tuberculosis developed. DISCUSSION: This case emphasizes the lack of specific clinical, radiological, endoscopic and histological signs of duodenal tuberculosis. PMID- 9205484 TI - [Asymptomatic carriers of vancomycin-resistant enterococci in France. Study in an ambulatory population of young subjects]. PMID- 9205485 TI - [Paraneoplastic cholestasis or Stauffer's syndrome. A new case]. PMID- 9205487 TI - [Abdominal pains and ascite in hemorrhagic fever with renal syndrome]. PMID- 9205486 TI - [Corticoids and HELLP syndrome. A new indicator?]. PMID- 9205488 TI - [Malignant non-Hodgkin's lymphomas. What indications for bone marrow autotransplantation?]. AB - Due to their chemosensitivity, intensive therapy with hematopoietic stem-cell autograft is widely used in the treatment of malignant non-Hodgkin lymphomas. Among the large body of literature on this topic, there are only a limited number of controlled trials. Bone-marrow graft is the standard treatment in lymphomas with highgrade malignancy when more than one factor of poor prognosis (elevated LDH, disseminated forms, poor general health) are observed at diagnosis and when only partial response is obtained after induction therapy. Similarly, the efficacy of bone-marrow grafts has recently been demonstrated to be greater than conventional treatment in case of relapse of chemosensitive forms. For other types of low-grade lymphoma and mantle-cell lymphoma, autologous bone-marrow graft has given very promising results but requires assessment in randomized studies to determine optimal use. PMID- 9205489 TI - [Integration of a psychiatric team to emergency care in catastrophe situation]. PMID- 9205490 TI - [Botulinum toxin: use in a case of achalasia of the lower sphincter of esophagus]. PMID- 9205491 TI - [Renal involvement in essential arterial hypertension]. AB - GENETIC DISEASE MODELS: A certain proportion of hypertension cases are due to renal disease. Recent advances in genetics has improved our knowledge of the pathophysiological mechanisms involved in certain rare diseases including apparent overproduction of mineralocorticoids, Liddle syndrome and Gitelman syndrome, and to hypothesize on the mechanism of primary hypertension. EFFECT ON PROGNOSIS: Onset of renal disease in hypertensive patients, whether expressed by proteinuria or the early stages of renal failure, worsens cardiovascular prognosis. FREQUENCY OF RENAL DISEASE: Renal disease is relatively rare in hypertensive patients, but as the general hypertensive population becomes older, there is a considerable rise in the prevalence of hypertensive renal disease as the underlying cause leading to dialysis. The risk of progressing to renal failure appears to be related to the level of the blood pressure, especially systolic pressure, at disease onset. Hypertension black subjects have a higher risk of developing chronic renal failure. THERAPEUTIC BENEFIT: Several studies have shown that lowering blood pressure with antihypertensive drugs lowers the risk progressing with primary hypertension. PMID- 9205492 TI - [Nephropathies and arterial hypertension]. AB - HIGH PREVALENCE: Hypertension is observed in 85% of patients with end-stage renal disease. Its prevalence is also very high in renal transplant recipients. POSSIBLE MECHANISMS: High blood pressure in primary renal disease may be related to water-electrolyte overload resulting from reduced sodium excretion (volume- or sodium-dependent mechanism). It could also result from renin-dependent mechanisms as in patients with unilateral disease or polycystic kidneys. PROGNOSIS: High blood pressure is a factor predicting poor prognosis in renal disease. It has been demonstrated that controlling hypertension has a beneficial effect on the course of renal failure. OTHER FACTORS: Hyperfiltration and high glomerular capillary pressure also play a role in the progression of chronic renal disease. The polymorphism of the angiotensin converting enzyme gene is also involved. In the diabetic patient as well as in patients with renal disease other than diabetes, a large amount of work has shown that converting enzyme inhibitors have more effect on showing the progression of renal failure than do other antihypertensive drugs. PMID- 9205493 TI - [Treatment of arterial hypertension with renal involvement]. AB - TREATMENT GUIDELINES: To prevent the development of hypertensive renal disease of progression of underlying renal disease, the blood pressure goal should be 130/85 mmHg or less if possible when proteinuria exceeds 1 g/24 h. ANTIHYPERTENSIVE DRUGS: Converting enzyme inhibitors are recommended in patients with non-diabetic renal disease and, as first-line therapy, in diabetics with microalbuminurea or patent diabetic nephropathy with or without hypertension. TREATMENT ONSET: Converting enzyme inhibitors should be started progressively with a lower final dosage in case of renal failure. Creatinemia and kaliemia must be monitored. Combination therapy with diuretics is generally used, but potassium-sparing diuretics are contraindicated due to the risk of hyperkaliemia. Reduced sodium intake is essential to obtain maximum effect. PMID- 9205494 TI - [Acoustic neuroma]. PMID- 9205495 TI - Gender and cutaneous melanoma. AB - Current evidence suggesting that a patient's sex is relevant to the progression of cutaneous melanoma is largely epidemiological. Although databases of patients with melanomas have for many years shown a survival advantage for female patients with primary melanoma, it has been difficult to evaluate contribution of other known prognostic variables such as thickness and site of the primary tumour, factors which also tend to be related to sex. In addition, there are data from a limited number of experimental studies and clinical trials which support the concept of female survival superiority in melanoma. This paper attempts to summarize the evidence for gender being an important factor in melanoma survival. PMID- 9205496 TI - Overexpression of protein kinase C-alpha and -beta isozymes by stromal dendritic cells in basal and squamous cell carcinoma. AB - Protein kinase C (PKC) isoenzymes transduce signals from cell surface receptors and thereby regulate important cellular functions in skin including keratinocyte growth and differentiation. Overexpression of individual PKC isoenzymes results in aberrant cell growth and in certain instances tumorigenicity. PKC is implicated in tumour promotion in mouse skin. Abnormal expression of PKC has been reported in several human cancers. We have, therefore, investigated expression of PKC-alpha and -beta in basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) by immunohistochemistry. Sections were stained with specific antibodies to PKC-alpha, PKC-beta, CD1a, T cells, B cells and dermal dendritic cells (factor XIIIa), using an immunoperoxidase technique. PKC-alpha and PKC-beta were not detected in tumour cells in BCCs or SCCs. In SCCs, PKC-beta immunostaining revealed positively stained inflammatory and dendritic cells scattered through the stroma; PKC-alpha immunostaining was essentially negative. In contrast, in BCCs, PKC-alpha+ and PKC-beta+ dendritic and spindle-shaped cells were observed in the stroma, immediately adjacent to the tumour islands. Double-labelling experiments showed that a proportion (approximately 20%) of PKC-beta+ dendritic cells also expressed factor XIIIa. BCCs depend on stroma for growth; PKC regulates expression of type IV collagenase and stromelysin III and interactions between tumour and stroma may be important in determining tumour invasion and metastasis. Therefore, overexpression of PKC-alpha and -beta by stromal dendritic cells in BCCs suggests that PKC activation may be involved in stromal/tumour interactions in these tumours. PMID- 9205497 TI - A recurrent laminin 5 mutation in British patients with lethal (Herlitz) junctional epidermolysis bullosa: evidence for a mutational hotspot rather than propagation of an ancestral allele. AB - The three genes (LAMA3, LAB3 and LAMC2) that encode the anchoring filament protein, laminin 5, may all harbour pathogenetic mutations in the autosomal recessive blistering skin disorder, junctional epidermolysis bullosa (JEB). Recently, one particular mutation, R635X in the LAMB3 gene, has been found to account for approximately 40% of all JEB laminin 5 mutations (Kivirikko et al., Hum Mol Genet 1996; 5: 231-7). In this study, we assessed the frequency of this mutation in 12 British patients with lethal (Herlitz) JEB using PCR amplification of genomic DNA and restriction endonuclease digestion. The mutation R635X was fond in seven of 24 (29%) mutant alleles, confirming its relative frequency within the British gene pool. In addition, haplotype analysis using intragenic polymorphisms showed that the mutation arose on at least four different haplotype backgrounds, suggesting it represents a mutational hotspot rather than propagation of a common British ancestral allele. These findings support the hypermutable nature of this CpG dinucleotide and have implications in screening for laminin 5 gene mutations in British and other patients with JEB. PMID- 9205498 TI - DNA analysis indicates patient-specific human papillomavirus type 16 strains in Bowen's disease on fingers and in archival samples from genital dysplasia. AB - Human papillomavirus (HPV) type 16 is casually involved in the pathogenesis of anogenital cancer and has also been demonstrated in some patients with Bowen's disease (BD) on the fingers. From two women with HPV 16 in BD on the fingers, and in archival samples from genital dysplasia, collected as long as 26 years ago, the non-coding region of the virus was amplified by the polymerase chain reaction and sequenced. The HPV 16 DNA sequences found in the finger lesions and in the genital archival samples showed no diversities within single patients. Compared with an HPV 16R reference sequence, one patient showed a unique T nucleotide at position 78, whereas the other patient exhibited T and A nucleotides at positions 7193 and 7521, respectively. In one of the patients, the same strain of HPV 16 was found in a digital tumour 26 years after its clearance from the genital tract. DNA sequence analysis indicated patient-specific HPV 16 strains. Auto inoculation from the genital tract was favoured as a plausible explanation of why HPV 16 caused BD on the fingers. PMID- 9205499 TI - Molecular detection of Treponema pallidum in secondary and tertiary syphilis. AB - Treponema pallidum can be detected by conventional techniques such as dark-field microscopy, immunofluorescence or the rabbit infectivity test, in large numbers in the skin lesions of primary and early secondary syphilis. In the skin lesions of late secondary and tertiary syphilis, conventional techniques fail to detect spirochaetes in general, perhaps due to increasing degeneration and the disappearance of treponemal spirochaetes in late syphilitic skin lesions. We used the highly sensitive technique of polymerase chain reaction (PCR) to prove the presence of Treponema pallidum-specific DNA in six lesions of late secondary syphilis and seven lesions of tertiary syphilis, including one syphilitic gumma. A Whartin-Starry stain was carried out in all 13 specimens and did not reveal any treponemal structures. Treponema pallidum-specific DNA was amplified by PCR in four of six cases of secondary syphilis and in the syphilitic gumma. These results are in favour of a direct cell-mediated immune reaction directed against treponemal antigen rather than the concept of an Id-reaction. Beside the usefulness of a PCR-based assay for understanding the aetiology of lesions of late syphilis, the assay described can be of clinical importance in various situations where traditional methods fail to detect Treponema pallidum because of lack of sensitivity. PMID- 9205500 TI - In situ evidence that the population of Langerhans cells in normal human epidermis may be heterogeneous. AB - Epidermal Langerhans cells (LC) may occur in subsets with different phenotypic and functional characteristics. In this work give further evidence that the CD1a positive LC population in the normal human epidermis may be heterogeneous. We found that one of our monoclonal antibodies (TE4B) to stratum corneum chymotryptic enzyme (SCCE) stained a population of dendritic cells in the normal epidermis in addition to high suprabasal keratinocytes. The staining of the dendritic cells was seen only when the biopsies had been fixed with formaldehyde and when the sections had been pretreated, either with proteolytic enzymes or with Triton X-100. The binding of the antibody was mediated through its antigen binding site, as it could be inhibited by adsorption with recombinant pro-SCCE. Experiments with double labelling showed that the TE4B-positive dendritic cells were also CD1a-positive. On the other hand, not all CD1a-positive cells were TE4B positive. By means of confocal microscopy of double-labelled cells, the TE4B binding site could be localized intracellularly. SCCE-mRNA could be detected by in situ hybridization in high suprabasal keratinocytes only. A possible explanation may be that there is a subset of LC which have taken up SCCE secreted by high suprabasal keratinocytes. Alternatively, TE4B may bind to an epitope present in a subgroup of epidermal LC which cross-reacts immunologically with SCCE. It is suggested that the demonstrated heterogeneity of the population of LC in the normal epidermis should be taken into account in studies on the possible role of epidermal autoantigens in the development of immune-mediated skin diseases. PMID- 9205501 TI - Anti-BP180 autoantibodies as a marker of poor prognosis in bullous pemphigoid: a cohort analysis of 94 elderly patients. AB - The prognosis of bullous pemphigoid (BP), a disorder which usually affects elderly patients, is not well established and conflicting data have been reported about the mortality rate of the disease. Our objective in this study was to assess the clinical and immunological factors determining survival in a prospective series of 94 patients with BP. A cohort of 94 consecutive patients with BP (mean age +/- SD: 81 +/- 4 years) was studied over an 8-year period (1987 94) in one department and patients followed up for at least 1 year. The diagnosis of BP was made on clinical criteria (using a standardized questionnaire), direct immunofluorescence (IF) findings (i.e. linear deposits of IgG and/or C3 along the basement membrane zone) and confirmed by direct immunoelectron microscopy and/or Western immunoblotting. Our analysis (median duration of follow-up: 5 years) showed that 37% of BP patients were dead within a year of starting treatment. The clinical or immunological factors which may influence the prognosis of BP were studied according to the criterion of death or survival by the end of the first year of treatment. None of the following factors was found to be significantly linked to the prognosis in BP: age, sex, extent of skin lesions at presentation, presence of mucosal lesions, blood eosinophilia, or the presence of circulating basement membrane zone autoantibodies by indirect IF. An impaired general condition and a history of coronary artery disease indicated a bad prognosis. The presence of circulating autoantibodies against BP180 autoantigen but not autoantibodies against BP230, as detected by immunoblotting on epidermal extracts, was found to be significantly more frequent (60% vs. 25%) in BP patients who died within the first year of treatment (P < 0.01). We conclude that the presence of circulating autoantibodies against BP180 represents the first intrinsic prognostic factor that has been demonstrated in BP. This result supports the growing body of evidence for the pathophysiological importance of the anti-BP180 autoantibodies. PMID- 9205503 TI - Cutaneous periarteritis nodosa: a clinicopathological study of 79 cases. AB - Cutaneous periarteritis nodosa (PAN) is a well-recognized entity characterized by tender subcutaneous nodules and livedo that may ulcerate. The pathogenesis of cutaneous PAN is not known. The objective of the study was to evaluate the clinical and histological features of 79 cases of cutaneous PAN and to investigate any clinical, pathological and immunological differences that may distinguish those cases likely to have a prolonged course. A retrospective analysis of 79 cases was conducted. Thirty-nine patients had ulcers during the course of their illness. Women were affected more than men. Painful nodules on the lower extremities, with oedema and swelling, were the most common clinical finding; 22% of patients had some evidence of neuropathy. Most of the laboratory findings were non-specific. There was no evidence for hepatitis B infection and hepatitis C infection was present in only one patient. Most patients (60%) had no associated medical condition. The disease course was prolonged but benign, and systemic PAN did not develop in any patient. Corticosteroids given systemically induced remission in most acute cases. The ulcerative form of disease was more prolonged and frequently associated with neuropathy. PMID- 9205502 TI - Photosensitivity in lupus erythematosus, UV photoprovocation results compared with history of photosensitivity and clinical findings. AB - Photosensitivity, one of the presenting symptoms in lupus erythematosus (LE), is still poorly defined and varying prevalence figures have been reported. The possibility of a coexisting photodermatosis, especially polymorphous light eruption (PLE), has often not been taken into account. We report the results of ultraviolet A (UVA) and B (UVB) photoprovocation tests in 67 clinically photosensitive patients who had confirmed discoid LE (DLE), systemic LE (SLE) or subacute cutaneous LE (SCLE). The results are compared with a detailed history of photosensitivity and with clinical and serological findings. A pathological photoprovocation reaction, graded as weak, moderate or strong, was induced with either UVA or UVB in 69% of patients with LE, in 100% of those with SCLE, in 70% of those with SLE and in 64% of those with DLE, but in none of 14 controls. Only 16% of the pathological reactions were strong and long-lasting, resembling LE lesions, while 48% were moderate or weak and transient, clinically like PLE. Fifty-three per cent of the provocation reactions which were biopsied showed a PLE-like histology or a non-specific inflammatory reaction, and most of them were clinically moderate or weak reactions of short duration. In the remaining, mostly clinically strong or long-lasting reactions, the histology was consistent with LE. A history of sunlight sensitivity did not predict a pathological photoprovocation result but a positive association between the presence of SSA/Ro or SSB/La antibodies and a pathological photoprovocation reaction was found. We have shown that PLE coexists with LE and that both PLE- and LE-like lesions can be induced with UV radiation in LE patients. PMID- 9205504 TI - Prediction of minimal erythema dose with a reflectance melanin meter. AB - The relationship between skin pigmentation and sensitivity to ultraviolet (UV) radiation-induced erythema was investigated in 60 healthy subjects of sun reactive skin types I-V. Using a portable reflectance spectrometer, skin pigmentation was measured as the 'melanin index' (MI) in all subjects. A solar simulated array of filtered UVA and UVB-emitting fluorescent lamps was then used to determine the UVB minimal erythema dose (MED) of each subject. MI readings and MED testing were both performed on the subjects' mid to upper backs. Using this technique, we found a close correlation between MI and MED. Comparison of the mean MI or MED of subjects with different skin types revealed progressive differences in MI and MED between all five skin types. Erythema dose-response curves, which provide further information about UV sensitivity, were also calculated for 43 subjects. A significant negative correlation was found between the gradients of these curves and both MI and MED, indicating that paler subjects respond more strongly to increments of UV above the MED than subjects with greater pigmentation. Our results indicate that although traditional, subjective means of predicting UV sensitivity to erythema are not without some value, MED correlates particularly strongly with objective measures of skin pigmentation. We therefore conclude that the reflectance spectrometer can rapidly and accurately predict UVB sensitivity, and should prove clinically useful for planning and optimizing UVB phototherapy. PMID- 9205505 TI - A study of the impact of Sun Awareness Week 1995. AB - We report a questionnaire-based survey of parents of schoolchildren to assess the impact of Sun Awareness Week, organized by the U.K. Skin Cancer Working Party, from 4 to 10 June 1995. A questionnaire was designed, pre-tested and modified. Nine hundred and eighty-nine patients of schoolchildren were approached before and after Sun Awareness Week at eight comprehensive primary schools and a children's clothes shop. Responses to the questionnaires were analysed in the following categories: total knowledge, sunscreen knowledge, melanoma risk factor knowledge, attitude, self-reported behaviour and awareness of the link between childhood sun exposure and cancer (link). A reasonable level of knowledge was demonstrated in the sample population. Significant improvements in attitude and behaviour scores were observed (P < 0.05). Three of the four link questions were significantly improved (P < 0.005). These results were not explained by differences in age, sex, or occupation between the before and after populations. Further national studies are needed to establish long-term benefits of such interventions. PMID- 9205506 TI - Adverse reactions following pulsed tunable dye laser treatment of port wine stains in 701 patients. AB - The pulsed tunable dye laser (PTDL) is generally considered to have a very low incidence of adverse effects, allowing it to become the treatment of choice for the majority of port wine stains (PWS). The low incidence of adverse effects has led to difficulties in determining the true incidence and type of adverse effect seen with this laser. We therefore undertook a retrospective study of 701 patients with PWS, who received 3877 full treatments to determine the incidence and type of adverse effects seen following treatment with the PTDL. Blistering and crusting were seen in 5.9% an 0.7% of patients, respectively, but were transient events which usually healed without permanent sequelae. Hyper pigmentation was the most frequently observed adverse effect seen in 9.1% of patients but generally showed gradual resolution over 6-12 months. Hypopigmentation was infrequent, seen in 1.4% of patients. The most significant adverse effects were atrophic and hypertrophic scarring seen in 4.3% and 0.7% of patients, respectively. Our observations show that there is a small but definite risk of atrophic scarring with a predisposition for younger patients. Hypertrophic scarring can occur albeit rarely and there may be predisposition towards the neck. In most cases test areas were not predictive of scarring. This underlines the need for a full discussion of scarring risk in patients with PWS undergoing treatment with the PTDL. PMID- 9205507 TI - IgG subclasses specific to Staphylococcus epidermidis and Propionibacterium acnes in patients with acne vulgaris. AB - IgG subclasses specific to Staphylococcus epidermidis and Propionibacterium acnes were determined in sera from patients with mild, moderate or severe acne and from a control group. Titres specific to S. epidermidis were all within the same range and did not differ between groups. The titres of IgG subclasses to specific to P. acnes did vary between groups. IgG1 and IgG3 were significantly higher in severe acne patients compared with moderate acne patients, while IgG2 was significantly higher in moderate and severe patients compared with controls. Titres of IgG4 did not differ between groups. The pattern of titres observed suggests that, while the antibody response to S. epidermidis is relatively harmless, antibodies to P. acnes may be involved in the pathogenesis of acne vulgaris. PMID- 9205508 TI - An investigation into the effect of ischaemia and pressure on irritant inflammation. AB - Reports of chemical burns beneath tourniquets during orthopaedic procedures led us to explore the irritant effects produced by the skin antiseptics used during such procedures. A sphygmomanometer and tourniquet, at a pressure of 200 mmHg for 30 min, was used to created pressure and ischaemia which were then examined separately for their respective effects on irritant inflammation in normal subjects and those with atopic eczema. As no inflammation could be demonstrated with the antiseptics, we subsequently used the known irritant chemical anthralin to examine the effect of ischaemia with and without pressure. Site-related variation in anthralin-induced inflammation was observed but there was no demonstrable effect of either pressure or ischaemia on the inflammatory response. Therefore, as we are unable to show a relationship between ischaemia with or without pressure and irritant inflammation, we conclude that burns under tourniquets are likely to be idiosyncratic reactions and their further investigation requires examination of the individuals affected. PMID- 9205509 TI - Evaluation of 6 weeks treatment of terbinafine in tinea unguium in a double-blind trial comparing 6 and 12 weeks therapy. The Lagos V Study Group. AB - Terbinafine (Lamisil) has been registered throughout the world for the treatment of finger and toenail onychomycosis. The recommended duration of treatment of toenail onychomycosis based on phase III studies is 12 weeks. This study was designed to determine: (i) if patients in whom the proximal part of the toenails was not affected respond as well after 6 weeks treatment as after 12 weeks treatment; (ii) to identify factors which may allow selection of patients for shorter treatment duration; and (iii) confirm that 6 weeks therapy is sufficient in fingernail mycosis. One hundred and forty-eight patients received 250 mg terbinafine daily for either 6 or 12 weeks in a double-blinded manner, and were allowed until 48 weeks after start of therapy. Cure of the nail infection was defined as negative mycological tests (mycological cure) and progressive growth of normal nail (clinical cure). Mycological cure was recorded in 43 of 72 (59.7%) in the 6-week group and 55 of 76 (72.4%) in the 12-week group. In those who completed the study per protocol in the 6-week group, 34 of 61 (55.7%) were cured mycologically corresponding to 46 of 56 (82.1%) in the 12-week group. The overall clinical and mycological cure rates for the two groups were 28 of 61 (45.9%) and 33 of 56 (58.9%), respectively. In the small number of patients with associated fingernail infection, all were improved and six of eight (75.0%) were cured after a duration of treatment of 6 weeks. A priori risk factors for failure of cure could not be identified in either group. However, shorter duration of disease prior to treatment and no involvement of the big toenail was associated with a trend toward better responses in both groups. It can be concluded from this study that, in toenail mycosis without visible matrix involvement, 6 weeks treatment of terbinafine is generally not sufficient, whereas fingernail infections respond well to this short therapy. PMID- 9205510 TI - Laser beam microdissection in the diagnosis of cutaneous B-cell lymphoma. AB - Interpretation of molecular analyses of cutaneous lymphoid infiltrates may be difficult because a heterogeneous group of cells in usually present within the neoplasms. Extraction of DNA from tissue sections does not provide exact information about which cell population has been analysed. We present a laser microscope system that allows selective molecular analysis of single cells or small groups of cells in cases of cutaneous lymphoma. An ultraviolet (UV)-laser microscope system (PALM, Wolfratshausen, Germany) was used to isolate particular populations of cells from a routinely processed specimen of a cutaneous follicular lymphoid proliferation. Using the UV-laser beam, a circle was cut around a target germinal centre in order to separate it from neighbouring tissues and to isolate a pure population of germinal centre cells. Isolated cells were scraped off with a micromanipulator and placed in a proteinase-K solution. DNA was extracted and amplified by the polymerase chain reaction (PCR) technique. Analysis of immunoglobulin JH gene rearrangement showed a distinct monoclonal band. In a second phase, using the same procedure in the same specimen, mantle zone cells around a germinal centre and single interfollicular B lymphocytes were isolated for PCR analysis of immunoglobulin JH gene rearrangement. In this population of cells, no clonality could be detected. This new technique allows the selective elimination of undesired cells and tissue from cutaneous neoplasms. By destruction of unwanted tissues with laser-beam energy a contamination-free sample is obtained. Analysis of isolated cells in our case demonstrated a clonal rearrangement derived from germinal centre cells and not from other B lymphocytes in the specimen, confirming the diagnosis of cutaneous follicle centre lymphoma. The method described has exciting implications for dermatology and dermatopathology, allowing precise correlation of morphological features with findings by molecular genetics. PMID- 9205511 TI - Molecular diagnosis of deep nodular bacillary angiomatosis and monitoring of therapeutic success. AB - A 51-year-old human immunodeficiency virus (HIV)-positive male patient (CDC stage 3C) had had a painful nodule on his external ankle joint for 10 months. A biopsy suggested bacillary angiomatosis, but Kaposi's sarcoma could not be excluded. Rods were detectable in lesional skin by a Warthin-Starry stain. A 298 base pair (bp) gene fragment specific for Bartonella species was amplified from lesional skin and direct nucleotide sequence analysis of the amplification product clearly identified Bartonella quintana. Kaposi's sarcoma-associated herpes virus specific DNA was not amplifiable by polymerase chain reaction (PCR) in our patient, suggesting that the lesion represented bacillary angiomatosis alone, despite clinical and histopathological features which suggested the coexistence of bacillary angiomatosis and Kaposi's sarcoma. The lesion regressed after erythromycin was prescribed. However, 4 and 9 weeks after initiation of therapy, PCR still yielded a positive result in material obtained by a swab. After complete healing, following 12 weeks of antibiotic therapy, PCR became consistently negative. The optimal length of antibiotic treatment in HIV-positive patients with bacillary angiomatosis is not yet known and inadequate therapy may be followed by disseminated disease and a fatal outcome. PCR-based monitoring of the success of treatment is valuable for determining the duration of treatment resulting in a cure. PMID- 9205512 TI - Angiosarcoma of the scalp associated with renal transplantation. AB - A case of cutaneous angiosarcoma occurring in a 51-year-old male renal transplant patient is reported. Multiple violaceous nodules surrounded by poorly demarcated red to purple discoloration were found on his scalp. Immunosuppressants consisting of azathioprine and prednisolone had been administered during the 12 year period since the renal transplantation. We diagnosed the lesion clinically as a cutaneous angiosarcoma and performed a wide surgical excision. The histopathological findings confirmed the diagnosis and showed tumour cells in the peripheral margin. Postoperatively, the patient started immunotherapy with systemic administration of recombinant interleukin-2 (rIL-2), but he refused to continue it because of the acute rejection of the transplanted kidney induced by the rIL-2. Instead he received radiation therapy (total 7000 rad) of the scalp. Although no recurrence was noticed for 15 months after the completion of radiation, he died due to lung metastasis from angiosarcoma. We review the seven cases, including ours, of angiosarcoma after renal transplantation that are reported in detail in the literature. PMID- 9205513 TI - Mosaic expression of uncein and 180-kDa bullous pemphigoid antigen in generalized atrophic benign epidermolysis bullosa. AB - Immunofluorescence microscopy of epidermodermal junction components in serial cryosections from the perilesional skin of a patient with generalized atrophic benign epidermolysis bullosa (GABEB) showed broken line-like staining of both BPAG2 (180-kDa bullous pemphigoid antigen) and uncein (antigen of 19-DEJ-1 monoclonal antibody), whereas integrin alpha 6 and laminin 5 were continuously expressed along the basement membrane zone. Immunoelectron microscopy revealed a mosaic distribution of the BPAG2/uncein positive and negative cells. BPAG2, a candidate protein of GABEB, probably has a close connection with uncein, and anchoring filament component. PMID- 9205514 TI - Nikolsky's sign: is it 'dry' or is it 'wet'? AB - Nikolsky's signs refers to the ability to induce peripheral extension of a blister as a consequence of applying lateral pressure to the border of an intact blister. Although initially used in reference to the pemphigus group of blistering dermatoses, a positive Nikolsky's sign can be seen in other bullous diseases such as toxic epidermal necrolysis and staphylococcus scalded skin syndrome. Appreciating whether the blister is 'wet' or 'dry' at the site of a positive Nikolsky's signs may have both diagnostic and prognostic significance which I illustrate with several clinical cases. Lastly, I review the significance of a positive Nikolsky's sign. PMID- 9205515 TI - Dermatomyositis with a pityriasis rubra pilaris-like eruption: a little-known distinctive cutaneous manifestation of dermatomyositis. AB - A pityriasis rubra pilaris-like eruption has been described in patients with dermatomyositis. These patients showed generalized follicular hyperkeratosis and diffuse thickening of the palms and soles. Histopathological findings consisted of keratotic plugging of the follicular infundibulum and features of erector pili myositis. We report on an 18-year-old woman with dermatomyositis. The diagnosis was established by characteristic enzymatic alterations, electromyographic pattern of myositis and the findings in a muscle biopsy, although the patient had no evidence of muscular weakness during a follow-up of 14 years. She developed an erythematosus and squamous eruption associated with diffuse palmoplantar keratoderma. Histopathological features consisted of a papillomatous epidermis with spicules of compact eosinophilic hyperkeratosis over the tips of papillae that were not related to hair follicles. Pityriasis rubra pilaris-like eruption seems to be a characteristic although uncommon cutaneous manifestation in dermatomyositis. PMID- 9205516 TI - The labial melanotic macule: a review of 79 cases. AB - Seventy-nine patients presented to the pigmented lesion clinic between 1983 and 1996 with labial melanotic macules. We have followed up these patients for up to 13 years and 3 months (mean 6 years, 3 months). We present evidence that this is a benign entity. The appropriate management is reassurance and discharge, with the advice to return only if the lesion grows or darkens. This did not occur in any of our patients during the time of their follow-up. PMID- 9205517 TI - Granulomatous contact dermatitis due to gold earrings. AB - A young female developed persistent nodules at sites of ear piercing with gold earrings and patch testing demonstrated a positive allergic response to gold sodium thiosulphate. Histological examination of the nodules demonstrated a prominent sarcoidal-type granulomatous tissue reaction. This is in contrast to previous reports of lymphocytoma cutis type histology and was associated with the occurrence of epithelioid granulomata at the site of a strongly positive and long lasting patch-test reaction. PMID- 9205518 TI - Linear IgA disease and ulcerative colitis. AB - Seventy adult patients with linear IgA disease were studied. Five (7.1%) also suffered from ulcerative colitis (the prevalence of ulcerative colitis in the U.K. is 0.05%). A further three patients with this association, from other British centres were also reviewed. In all cases, ulcerative colitis preceded the onset of skin disease, by a median of 6.5 years. The reason for this association remains unclear but the abnormalities in colonic mucosal B cells and mucosal IgA1 production reported in patients with ulcerative colitis may be important in the subsequent development of IgA1 mediated linear IgA disease. PMID- 9205519 TI - Necrolytic migratory erythema and zinc deficiency. AB - Necrolytic migratory erythema (NME) is an uncommon condition classically associated with high plasma levels of circulating glucagon and a glucagonoma. We report a patient with cirrhosis who showed clinical and histological features of NME. Investigation revealed normal glucagon levels without evidence of glucagonoma. Serum zinc levels were below the normal range and zinc supplementation resulted in rapid and complete resolution of the eruption. PMID- 9205520 TI - A higher prevalence of onychomycosis in psoriatics compared with non-psoriatics: a multicentre study. AB - There is some controversy about the prevalence of onychomycosis in patients with psoriasis compared to non-psoriatics. We therefore measured the prevalence of toenail onychomycosis in psoriatics and non-psoriatics attending dermatologists' offices. None of the patients had a referring diagnosis of onychomycosis. The prevalence of pedal onychomycosis in psoriatics (n = 561) was 13%. The odds of patients with psoriasis having onychomycosis was 56% greater than non-psoriatics of the same age and sex (P = 0.02). In the psoriatics, when the toenails were clinically abnormal, the prevalence of onychomycosis was 27%. The odds of developing onychomycosis increased with age (P < 0.0001) and the odds of men developing onychomycosis was 2.5 times that of women (P = 0.0001). The duration of psoriasis did not significantly affect the odds of developing onychomycosis. The fungal organisms recovered from psoriasis subjects with onychomycosis were similar to those in the normal population with onychomycosis (P = 0.58). PMID- 9205521 TI - Urinary leukotriene E4 levels in patients with atopic dermatitis. AB - Leukotriene synthesis may be increased in a variety of inflammatory diseases. Urinary leukotriene E4 is a stable metabolite of leukotrienes C4 and D4 which has previously been found to be increased in exacerbations of severe asthma and after antigen inhalation. Levels of urinary LTE4 in seven patients during and after a severe flare of atopic dermatitis were measured by high-performance liquid chromatography (HPLC) and radioimmunoassay (RIA). Mean urinary LTE4 levels (+/- SEM) were not increased during (16.7 +/- 3.7 pg/mumol) or after (16.9 +/- 4.8 mumol) the acute exacerbation of atopic dermatitis when compared with the normal range (mean = 23.8 [95% confidence interval 19.9-28.2] pg/mumol creatinine). These findings do not provide evidence of cysteine leukotriene involvement in the pathogenesis of atopic dermatitis. PMID- 9205522 TI - Keratoderma simplex: a novel entity simulating seborrhoeic keratosis. PMID- 9205523 TI - Leishmaniasis presenting as a psoriasiform eruption in AIDS. PMID- 9205524 TI - Vesicular pemphigoid with antidesmoplakin autoantibodies. PMID- 9205525 TI - Juvenile nodulocystic acne responding to systemic isotretinoin. PMID- 9205526 TI - Actinic granulomata in a young woman following prolonged sunbed usage. PMID- 9205527 TI - Isotretinoin-PUVA in women with psoriasis. PMID- 9205528 TI - Ultraviolet radiation and primary school children. PMID- 9205529 TI - 8-Methoxypsoralen PUVA for psoriasis: a comparison of a minimal phototoxic dose based regimen with a skin-type approach. PMID- 9205530 TI - Serological screening for coeliac disease in vitiligo and alopecia areata. PMID- 9205531 TI - Endocrine factors in pre- and postmenopausal women with hidradenitis suppurativa. PMID- 9205532 TI - p21Waf1/Cip1 expression. PMID- 9205533 TI - Human herpesvirus 8 DNA is rarely found in Bowen's disease of non immunosuppressed patients. PMID- 9205534 TI - DNA diploidy in AIDS-related and steroid-induced Kaposi's sarcoma. PMID- 9205535 TI - Food-dependent exercise-induced anaphylaxis due to matsutake mushrooms. PMID- 9205536 TI - Cyclosporin treatment of nodular prurigo in a dialysis patient. PMID- 9205537 TI - von Willebrand factor: a marker of endothelial dysfunction in vascular disorders? AB - The vascular endothelium is involved in the production of many important substances which are involved in a cardiovascular pathophysiology. One such substance which is synthesised by, and stored in, endothelial cells is von Willebrand factor (vWf). When released, vWf appears to mediate platelet aggregation and adhesion. Numerous clinical and experimental reports suggest that high vWf levels reflect damage to the endothelium or endothelial dysfunction. The close association between vWf and the processes of thrombus formation (thrombogenesis) or atherogenesis also suggests that high vWf levels may be a useful indirect indicator of atherosclerosis and/or thrombosis. The availability of a useful marker of endothelial dysfunction may have potential clinical value. The measurement of such a marker can perhaps be a non-invasive way of assisting in clinical diagnosis or as an indicator of disease progression. High vWf levels have also been shown to have prognostic value in patients with ischaemic heart disease, peripheral vascular disease and inflammatory vascular disease. However, there is limited information that increased vWf is actually casual in the progression of vascular disease and that measures aimed at reducing vWf levels will be beneficial. In addition, interpretation of raised plasma vWf levels is complicated by the fact that vWf may be an acute phase reactant. Further research is indicated to explore the predictive value of this marker in population studies and, perhaps, therapeutic approaches in (and the value of) modifying vWf levels or function. PMID- 9205538 TI - Herbs, seeds, oil and eggs: a vasotoxic salad. PMID- 9205539 TI - Pulmonary hypertension: updating a mysterious disease. PMID- 9205541 TI - Right ventricular dysfunction persists following brief right ventricular pressure overload. AB - OBJECTIVE: Acute pulmonary hypertension may cause right ventricular (RV) contractile failure. While it has been assumed that restoration of normal loading conditions after acute pulmonary hypertension is sufficient for complete recovery of RV function, this has not been rigorously examined. The purpose of this study was to test the hypothesis that acute RV pressure overload produces RV contractile dysfunction that persists following restoration of control loading conditions. METHODS: We subjected 18 autonomically-blocked, chloralose anesthetized, open-chest pigs to 1 h of pulmonary artery constriction to increase RV systolic pressure from 35 +/- 1 to 55 +/- 1 mmHg, followed by 2 h of measurements after pulmonary artery constriction release. We determined regional RV free wall function from pressure-segment length loops and preload recruitable stroke work relations, and global RV function from stroke work vs. end-diastolic pressure relations. RESULTS: As expected, RV free wall systolic shortening diminished during pulmonary artery constriction, but the endo/epi blood flow ratio, lactate uptake, and coronary venous pH were not significantly changed. Following release of pulmonary artery constriction, RV systolic and diastolic pressure returned to control values. Nonetheless, contractile dysfunction persisted, with depressed RV free wall systolic shortening (70 +/- 22% of control), RV regional external work (59 +/- 11% of control at control end diastolic length), and global RV stroke work (56 +/- 14% of control at control end-diastolic pressure). Depressed regional work was due to a parallel, rightward shift of the preload recruitable stroke work relation. Five pigs identically instrumented but not subjected to pulmonary artery constriction showed no significant over 3 h. CONCLUSIONS: Acute pulmonary hypertension causes RV contractile dysfunction that persists at least 2 h after restoration of control loading conditions. Contractile dysfunction is not attributable to RV ischemia during pressure overload. PMID- 9205540 TI - Acute and persistent effects of a 46-kilometer wilderness trail run at altitude: cardiovascular autonomic modulation and baroreflexes. AB - OBJECTIVE: To test the hypothesis that prolonged physical exercise induces long lasting effects on blood pressure and heart rate we studied 17 endurance runners before and after the 1995 Sandia Wilderness Crossing Research Run (46 km of rocky trails, average altitude 2500 m). METHODS: We evaluated the response of the cardiovascular system to sympathetic stimulation by orthostatism and to sympathetic and parasympathetic carotid baroreceptor stimulations by sinusoidal neck suction at different frequencies (sympathetic activity on blood pressure by low-frequency stimulation, parasympathetic activity on RR interval by high frequency stimulation). We used power spectral analysis of beat-to-beat RR interval, systolic and diastolic non-invasive blood pressure, in order to quantify the respiratory fluctuations (depending on vagal activity on the RR interval) and the slower non-respiratory fluctuations, depending on sympathetic activity on the blood pressure. Recordings were performed 24 h before, and 30 min, 24 h and 48 h after the run. RESULTS: Thirty minutes after the race we found reduced blood pressure, signs of relative sympathetic predominance (increased RR interval low-frequency/high-frequency ratio from 0.65 +/- 0.15 to 1.63 +/- 0.37, P < 0.05), reduced effect of parasympathetic baroreceptor stimulation (decrease in RR interval high-frequency neck-suction synchronous oscillations, from 5.33 +/ 0.34 to 3.55 +/- 0.37 ln-ms2, P < 0.005), unchanged blood pressure responses to sympathetic stimulations; 24 h after the race, the response to parasympathetic stimulation was increased (to 6.44 +/- 0.32 ln-ms2, P < 0.0005) compared to baseline (24 h before the race), whereas sympathetic stimulation by neck suction had no longer an effect on blood pressure. CONCLUSION: The acute effects of prolonged exertion are associated with a relative increase in sympathetic activity. Twenty-four hours after this race an increased sensitivity to vagal and reduced sensitivity to sympathetic baroreflex stimulation was found. In this field study at altitude we found long-lasting effects on cardiovascular autonomic modulation after physical exertion. PMID- 9205543 TI - The effects of left ventricular stretch versus cavity pressure on intramyocardial pressure. AB - OBJECTIVE: Since muscles, vessels and interstitial spaces are in close physical proximity in the heart wall, interstitial (i.e., intramyocardial) pressure (IMP) should be affected by the stresses of the vessels and/or the muscular tissue surrounding the interstitial spaces. Thus, we tested the hypothesis that increasing the stresses (or stiffness) of the surrounding tissues by muscle contraction or stretching--produced externally by stretching the LV cavity or internally by increasing coronary perfusion pressure--has a greater effect than LV cavity pressure per se on IMP. METHODS: In isolated rabbit hearts we measured IMP with small (< 10 microns diam) glass micropipettes while stretching the vessels (by changing coronary perfusion pressure) and the wall (by inflating a balloon in the left ventricle) during the passive state as well as during barium contracture. RESULTS: With LV cavity pressure equal to 0 (balloon open to air) or equal to 30 mmHg, a 20 mmHg increase in perfusion pressure increased IMP by 3.6 and 5 mmHg, respectively, in the passive state and by 7.6 and 7.9 mmHg, respectively, in the contracted state. This 30 mmHg increase in LV pressure produced a significant but small (3-5 mmHg) increase in IMP in the passive state but no effect in the contracture state. CONCLUSIONS: These results can be explained by a unifying concept in which stretching of the tissues surrounding the intestinal spaces--produced externally by increasing ventricular cavity size or internally by pressurizing vessels--but not LV cavity pressure per se is the major determinant of IMP. PMID- 9205542 TI - Insulin improves heart function and metabolism during non-ischemic cardiogenic shock in awake canines. AB - OBJECTIVES: This study was undertaken to examine in-situ heart function and metabolism during insulin treatment of verapamil-induced cardiogenic shock in awake canines. METHODS: Twenty mongrel canines were instrumented to monitor myocardial substrate uptakes (glucose, lactate, free fatty acids, oxygen [MVO2]), as well as ventricular (LV) end-systolic elastance (Emax), LV efficiency (LV minute work/MVO2), and Tau. Shock was induced by graded intraportal verapamil infusion followed by randomized assignment to one of 4 treatment groups: saline control (3.0 ml/kg/min, n = 5), epinephrine (5 micrograms/kg/min, n = 5), glucagon (10 micrograms/kg/min, n = 5) or insulin (1000 mU/min, n = 5) with dextrose to clamp arterial [glucose] +/- 10% of basal concentrations. RESULTS: Insulin treatment significantly increased Emax (34 +/- 3 vs. 17 +/- 3 mmHg/mm, saline control), and shortened Tau (9 +/- 3 ms) compared to saline control (42 +/ 5 ms), epinephrine (20 +/- 4 ms) and glucagon (35 +/- 8 ms). With insulin treatment, mechanical efficiency increased to 20,097 +/- 2070 vs. 12,424 +/- 1615 mmHg.mm/ml/O2/100 g in controls. Simultaneously, insulin increased myocardial lactate uptake (35 +/- 2 vs. 17 +/- 4 mumol/min/100 g. saline control), but did not increase glucose uptake. Epinephrine and glucagon decreased mechanical efficiency compared to saline controls, coincident with increased myocardial fatty acid consumption, but without increasing lactate uptake. One dog died early with glucagon treatment before the first death in the saline-treated group. CONCLUSIONS: Insulin improves systolic and diastolic heart function during aerobic shock and accelerates in-vivo myocardial lactate oxidation. PMID- 9205544 TI - Temporal dependence of coronary collateral development. AB - OBJECTIVE: Previous evidence suggests that episodes of myocardial ischemia of sufficient duration and intensity are required to produce coronary collateral development during repetitive coronary occlusion. This investigation tested the hypothesis that coronary collateral development is also temporal-dependent. METHODS: Chronically instrumented dogs (n = 16) were subjected to brief (2 min) left anterior descending coronary artery (LAD) occlusions, once every hour, 8 h a day, for 3 weeks or once every hour, 24 h a day for 1 week. Collateral perfusion (radioactive microspheres), LAD contractile function (ultrasonic crystals), and post-occlusive flow debt repayment (LAD flow probe) were measured during occlusions 1, 55, 105, and 155. RESULTS: Increases (P < 0.05 in subendocardial collateral blood flow to ischemic myocardium, progressive normalization of contractile function during LAD occlusion, and successive reduction in flow debt repayment were observed in dogs receiving occlusions over 3 weeks. In contrast, dogs receiving the same number of coronary occlusions over 1 week demonstrated minimal increases in collateral blood flow, persistent regional contractile dysfunction, and sustained flow debt repayment. CONCLUSIONS: The results demonstrate that LAD collateral development in response to repetitive coronary occlusion requires sufficient time for growth adaptation of the collateral circulation to occur. PMID- 9205545 TI - Protection by hypoxic preconditioning against hypoxia-reoxygenation injury in guinea-pig papillary muscles. AB - OBJECTIVE: Developed tension in guinea-pig papillary muscles is depressed by prolonged hypoxia; subsequent reoxygenation leads to a partial recovery that stabilizes after an early period of arrhythmia. We have investigated whether hypoxic preconditioning in these muscles (1) improves the recovery of developed tension, (2) protects against arrhythmia, and (3) causes other significant electromechanical changes. METHODS: Papillary muscles stimulated at 1 Hz were superfused with oxygenated Krebs solution for 60 min and either preconditioned (5 min of 3 Hz pacing substrate-free hypoxic conditions, 10 min of normoxic recovery) or equilibrated for an extra 15 min. Muscles were subsequently challenged with substrate-free hypoxia (1 Hz), and reoxygenated (1 Hz) for 60 min. Contractile performance, action potential parameters, and indicators of arrhythmic activity were measured in 10 preconditioned and 10 non-preconditioned muscles. RESULTS: Developed tension in preconditioned muscles declined to the same level (10-15% control) as in non-preconditioned muscles after 60 min hypoxia. A notable difference was that developed tension in the preconditioned muscles failed to rebound during mid-hypoxia, a hallmark feature in non preconditioned muscles. The action potential duration and overshoot collapsed at a significantly faster rate in hypoxic preconditioned muscles. Action potential recovery during reoxygenation was similar in the two groups of muscles, but recovery of developed tension was significantly stronger in preconditioned (76.7 +/- 5.4%) than in non-preconditioned (42.9 +/- 1.7%) muscles (P < 0.001). Reoxygenation provoked arrhythmic activity in all muscles, but the summed average duration was shorter (5.5 +/- 1.0 vs. 9.4 +/- 1.5 min) (P < 0.05) in the preconditioned muscles. CONCLUSION: Hypoxic preconditioning can significantly enhance post-hypoxia recovery of developed tension, and significantly attenuate arrhythmic activity, in guinea-pig papillary muscles. PMID- 9205546 TI - Late persistent expressions of ICAM-1 and VCAM-1 on myocardial tissue in children with lymphocytic myocarditis. AB - BACKGROUND: Both intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) have been implicated in cardiac allograft rejection. However, there is little information about the relationship between the expression of these adhesion molecules and myocarditis in children. METHODS AND RESULTS: Immunoreactivities of ICAM-1 and VCAM-1 were examined by enzyme immunoassay in 31 biopsy specimens obtained from 11 pediatric patients with biopsy-proven myocarditis or cardiomyopathy. Five of the 11 patients had clear evidence of acute myocarditis. The other 6 had ECG abnormalities identified by mass screening for heart disease, and subsequently had been histologically diagnosed as having non-specific cardiomyopathy. The period between onset of myocarditis or identification of ECG abnormality and immunohistochemical studies was 23 to 60 days and 8 months to 3 years, respectively. Expression of ICAM-1 and VCAM-1 was assessed by counting ICAM-1 and VCAM-1 positive vessels and dividing by the total number of vessels. ICAM-1 was significantly present on 81% (P < 0.01) of myocardial tissue samples in the 5 patients with healing-stage acute myocarditis, and on 45% (P < 0.05) in the remaining 6 patients with non-specific cardiomyopathy, compared with 24% in control specimens obtained from right ventricular muscle resected at surgery for tetralogy of Fallot. VCAM-1 was also present on 50% (P < 0.05) of the samples from the 5 patients with acute myocarditis, but was not present in those with non-specific cardiomyopathy. CONCLUSION: This persistent expression of ICAM-1 suggests that myocardial cell damage may persist immunologically for a long period in myocarditis. In addition, immunostaining for these adhesion molecules may be diagnostic value in clinically silent lymphocytic myocarditis and chronic cardiomyopathy. PMID- 9205547 TI - Short-term inhibition of the Na-H exchanger limits acidosis and reduces ischemic injury in the rat heart. AB - INTRODUCTION: Pharmacologic inhibition of the Na-H exchanger prior to and during ischemia has been shown to protect the ischemic heart by reducing Na-H exchange. However, pH regulation in the ischemic heart is primarily mediated by other pH regulatory mechanisms, such as metabolite efflux and sodium-coupled HCO3-influx, which may compensate for a reduction in Na-H exchange by increasing proton efflux. We hypothesized that short-term pharmacologic inhibition of the Na-H exchanger would result in increases in other compensatory pH regulatory mechanisms and thereby limit acidosis during ischemia and reduce ischemic injury. METHODS: In order to test this hypothesis, we exposed isolated perfused rat hearts to ethylisopropylamiloride (EIPA, 3 micro M) for 40 min, followed by 10 min of EIPA-free perfusate and 30 min of global ischemia (termed CTL/EIPA hearts). The effects of this intervention were compared to hearts perfused with either glucose alone (CTL) or EIPA 3 micro M for 10 min before ischemia (EIPA). Ischemic injury was measured using creatine kinase (CK) release on reperfusion, while pH and metabolic effects were measured using 31P nuclear magnetic resonance spectroscopy. The effect of this intervention on recovery from an acid load was assessed using an NH4Cl pre-pulse in bicarbonate-containing Krebs-Henseleit as well as a HEPES buffer. RESULTS: Both CTL/EIPA and EIPA hearts had marked reduction in ischemic injury (CK control 1191 +/- IU/g dry weight: CTL/EIPA 406 +/- 42 IU/gdw; EIPA 333 +/- 78 IU/gdw), as well as significantly reduced end diastolic pressure on reperfusion. Intracellular pH was higher in the CTL/EIPA hearts (end-ischemic pH = 6.34 +/- 0.05) compared to either control (5.86 +/- 0.02) or EIPA hearts (6.01 +/- 0.02), while pH recovery on reperfusion was markedly slowed in the CTL/EIPA hearts. CTL/EIPA hearts had rapid ATP depletion during ischemia, but PCr recovery comparable to EIPA hearts. Acidification on exposure to NH4Cl was increased in the presence of HEPES, but ph recovery was not altered by short-term exposure to EIPA. CONCLUSIONS: These data show that short term inhibition of the Na-H altered pH regulation in the ischemic heart, resulting in reduced acidosis and slow pH recovery on reperfusion, coupled with reduction in ischemic injury and end-diastolic pressure on a reperfusion. These findings are consistent with short-term exposure to EIPA accelerating ATP depletion during ischemia, as well as limiting proton efflux during reperfusion. PMID- 9205548 TI - Adenosine A2 receptor function in rat ventricular myocytes. AB - OBJECTIVE: This study was undertaken to investigate the functional significance of adenosine A2 receptor stimulation in a mammalian ventricular myocyte preparation. METHODS: Isolated contracting rat ventricular myocytes were employed to assess the contractile, adenylyl cyclase and cyclic AMP responses to adenosine receptor stimulation. RESULTS: In single myocytes the presence of A1 receptors was confirmed, as indicated by the A1 receptor agonist, phenylisopropyladenosine (PIA), reducing by 60 and 74% the inotropic response and activation of adenylyl cyclase, respectively, elicited by the beta-adrenergic agonist, isoproterenol. An A1 receptor antagonist, dipropylcyclopentylxanthine (DPCPX), prevented the antiadrenergic action of PIA. The A2 receptor agonist, carboxyethylphenethyl aminoethyl-carboxamido-adenosine (CGS-21680; 0.01-10 microM) increased myocyte inotropy in a concentration-dependent manner, reaching a maximum of 41-45%. Ethylcarboxamidoadenosine (NECA), naphthyl-substituted aralkoxy-adenosine (SHA 082) and adenosine in the presence of DPCPX also increased myocyte inotropy, as evidenced by increases in myocyte shortening, duration of shortening, time-to peak shortening, time-to-75% relaxation and rate of maximal shortening. The agonists, however, did not effect the maximal rate of relaxation. The A2 receptor antagonists, chlorofuranyldihydrotri-azoloquinazolinimine (CGS-15943) and chlorostyrylcaffeine (CSC), the latter selective for the A2a receptor, prevented the contractile responses elicited by the A2 agonists. Compared to the concentrations of A2 receptor agonists necessary to increase myocyte contractile variables, 3-12 times greater concentrations of the agonist were required to increase myocyte adenylyl cyclase activity and cAMP levels. CONCLUSIONS: The results suggest the presence of adenosine A2a receptors in the rat ventricular myocyte that appear to be responsible for an increase in inotropy via cAMP dependent and -independent mechanisms. PMID- 9205550 TI - Role of nitric oxide, cyclic GMP and superoxide in inhibition by adenosine of calcium current in rabbit atrioventricular nodal cells. AB - OBJECTIVE: To study the intracellular pathways which mediate the inhibitory actions of adenosine on isoprenaline-stimulated calcium current (ICa) in atrioventricular (AV) nodal myocytes. METHODS: The whole-cell patch-clamp technique was used to record ICa from rabbit AV nodal cells, isolated by enzymatic and mechanical dispersion. RESULTS: Isoprenaline, 0.1 microM, increased peak ICa from 0.58 +/- to 1.23 +/- 0.1 nA, and this increase was reversibly inhibited by adenosine, 10 microM (83 +/- 6%), which we have previously shown to be mediated by nitric oxide (NO) production. A membrane-permeable analogue of cyclic GMP, 8-Br-cGMP (300 microM), an inhibitor of cGMP-stimulated phosphodiesterase, prevented the effect of adenosine on ICa-Methylene blue (10 microM), an inhibitor of NO-sensitive guanylyl cyclase and a generator of superoxide (.02-), did not prevent, but increased, the inhibiting action of adenosine (49.5 +/- 6.6%, P < 0.01). Methylene blue (50 microM) caused a reduction of ICa, with further inhibition when combined with adenosine. A .O(2-) generating system, xanthine oxidase (0.02 U/ml) and purine (2.3 mM), also increased the inhibitory action of adenosine on ICa. Inhibition of ICa by adenosine in the presence of xanthine oxidase was not prevented by 8-Br-cGMP (300 microM) and was not influenced by pre-incubation of cells with a NO synthase inhibitor, L-NAME (0.5 mM). CONCLUSIONS: The inhibitory effect of adenosine on ICa in rabbit AV nodal myocytes can be mediated by two mechanisms--stimulation of cGMP-stimulated phosphodiesterase by NO-induced cGMP, and a mechanism which involves interaction with .O2- production. PMID- 9205549 TI - Similarities between early and delayed afterdepolarizations induced by isoproterenol in canine ventricular myocytes. AB - OBJECTIVES: This study aims at clarifying the role of cellular Ca2+ overload and spontaneous sarcoplasmic reticulum (SR) Ca2+ release in the generation of early afterdepolarizations (EAD) by isoproterenol. The involvement of a Ca(2+) activated membrane current in isoproterenol-induced EAD is investigated. METHODS: Membrane potential and contraction (an indicator of SR Ca2+ release) were recorded in canine left ventricular myocytes at pacing cycle lengths (CL) of 300 4000 ms. Threshold concentration for EAD was 20-50 mmol/l isoproterenol. Ni2+ (2.0-5.0 mmol/l) was used at normal and high (5.4 mmol/l) [Ca2+]o to examine the role of Ca2+ current and/or Na(+)-Ca2+ exchange (1Na-Ca) in EAD. RESULTS: In all cells delayed afterdepolarizations (DAD) appeared during isoproterenol. In most (approximately equal to 70%) cells EAD were also generated, which were fast pacing dependent, occurring only at CL of 400-1000 ms. EAD were always initiated by a delay in repolarization. Early aftercontractions preceded the EAD upstrokes, often occurring without them. They coincided with the initial delays in repolarization. During treatment with isoproterenol, Ni2+ and high [Ca2+]o, EAD and DAD were suppressed despite the continued presence of early and delayed aftercontractions. CONCLUSIONS: Our data indicate that beta-adrenergic EAD share a common ionic mechanism with DAD in terms of cellular Ca2+ overload and spontaneous SR Ca2+ release. beta-Adrenergic EAD consist of two phases: (1) a conditional phase coinciding with the onset of an early aftercontraction, often followed by (2) an EAD upstroke. A Ca2(+)-activated membrane current, probably I Na-Ca, is necessary at least for the initiation of these EAD. PMID- 9205551 TI - Expression and regulation of 80K/MARCKS, a major substrate of protein kinase C, in the developing rat heart. AB - OBJECTIVE: Protein kinase C (PKC) plays a pivotal role in modulating the growth and differentiation of many cell types including the cardiac myocyte. However, little is known about molecules that act immediately downstream of PKC in the heart. In this study we have investigated the expression of 80K/MARCKS, a major PKC substrate, in whole ventricles and in cardiac myocytes from developing rat hearts. METHODS: Poly A/ RNA was prepared from neonatal (2-day) and adult (42 day) cardiac myocytes and whole ventricular tissue and mRNA expression determined by reverse transcription-polymerase chain reaction (RT-PCR) using primers designed to identify a 420 bp fragment in the 80K/MARCKS gene. Protein extracts were prepared from either 2-day and 42 day cardiac myocytes or from whole ventricular tissue at 2, 5-11, 14, 17, 21, 28 and 42 days of age. Protein expression was determined by immunoblotting with an 80/MARCKS antipeptide antibody and PKC activity was determined by measuring the amount of gamma 32 P ATP transferred to a specific peptide substrate. RESULTS: RT-PCR analysis of 80K/MARCKS mRNA in neonatal (2-day) and adult (42-day) cardiac myocytes showed the expression of this gene in both cell types. Immunoblotting revealed maximum 80K/MARCKS protein expression in whole ventricular tissue at 5 days (a 75% increase above values at 2 days), followed by a transient decrease in expression during the 6-8 day period (61% of the protein expressed at 2 days for 8-day tissue) with levels returning to 5 day levels by 11 days of age. 80K/MARCKS protein was present in cardiac myocytes at 2 days of age whereas it was not detectable in adult cells. In addition, PKC activity levels increased to 160% of levels present at 2 days in 8-day old ventricles with PKC activity levels returning to 5-day levels by 9 days of age. This was then; followed by a steady decline in both 80K/MARCKS protein expression and PKC activity through to adulthood. CONCLUSIONS: Expression of the PKC substrate, 80K/MARCKS, in cardiac myocytes changes significantly during development and the transient loss of immunoreactive protein during the 6-8 day development period may reflect 80K/MARCKS phosphorylation and subsequent down-regulation as a result of the concomitant up-regulation of PKC activity at this time. PMID- 9205552 TI - Increased expression of promatrix metalloproteinase-9 and neutrophil elastase in canine dilated cardiomyopathy. AB - OBJECTIVE: Canine dilated cardiomyopathy, commonly affecting Doberman pinschers, results in extracellular matrix remodelling within the myocardium. The aim of this study was to examine the proteolytic activity in myocardium from Doberman pinschers with dilated cardiomyopathy. METHODS: Samples of myocardium, obtained rapidly post mortem from the left ventricular free wall of Dobermans with dilated cardiomyopathy, clinically normal Dobermans and control dogs (non-Dobermans), were examined for proteolytic activity using substrate gel zymography. Gels were analysed by scanning densitometry. RESULTS: Promatrix metalloproteinase-9 activity was significantly increased in all Doberman myocardium when compared to controls. A significant increase in an enzyme, identified to be neutrophil elastase by inhibition of its activity by Elastatinal and Western blotting, was also detected in all Dobermans when compared to controls. CONCLUSIONS: The results indicate that promatrix metalloproteinase-9 and neutrophil elastase, both of which are implicated in inflammatory responses, are present in significantly elevated levels in Doberman dilated cardiomyopathy and are raised in clinically normal Dobermans. Both proteolytic enzymes degrade a wide variety of connective tissue components and thus the increased levels found may play an important role in the structural remodelling seen in the myocardium and subsequent heart failure. Increased proteolytic enzyme levels in clinically normal Dobermans may be indicative of the predisposition of the breed to dilated cardiomyopathy. PMID- 9205553 TI - Sympathetic modulation of hypoxic pulmonary vasoconstriction in intact dogs. AB - BACKGROUND: The effects of the sympathetic nervous system on hypoxic pulmonary vasoconstriction (HPV) have been reported variably. We studied the effects of adrenergic receptor blockade and epidural blockade on HPV in 32 pentobarbital anaesthetised intact dogs. METHODS: Pulmonary arterial flow-pressure relationships were determined in hyperoxia and hypoxia, at baseline and after alpha-blockade (phentolamine 2 mg/kg + 50 micrograms.kg-1.-1), beta-blockade (propranolol 2 mg/kg), alpha beta-blockade, epidural blockade (lignocaine 20 mg/kg), and alpha beta-plus epidural blockade. RESULTS: At reference flow of 3.5 1.min-1.m-2, the mean hypoxic response (hypoxia-induced increase in transpulmonary pressure gradient, each n = 8) changed from 6.0 +/- 0.9 to 3.5 +/- 1.0 mmHg after alpha-blockade, from 5.8 +/- 0.9 to 0.7 mmHg after beta-blockade, from 4.1 +/- 0.8 to 0.9 +/- 1.4 mmHg after alpha beta-blockade from 3.4 +/- 1.0 to 4.3 +/- 0.9 mmHg after epidural blockade (all P < 0.05), and was not affected by epidural blockade after alpha beta-blockade. CONCLUSIONS: In pentobarbital anaesthetised dogs, (1) HPV is attenuated by alpha- and enhanced by beta-, alpha beta- and epidural blockade, and (2) epidural blockade has no significant adrenergic-unrelated effect on the pulmonary vasculature. PMID- 9205554 TI - Right ventricular angiotensin converting enzyme activity and expression is increased during hypoxic pulmonary hypertension. AB - OBJECTIVE: To determine whether local cardiac angiotensin converting enzyme (ACE) expression is upregulated during the development of hypoxia-induced right ventricular hypertrophy. METHODS: ACE activity was measured in membrane preparations from the right ventricle and left ventricle plus septum in normoxic rats and animals exposed to chronic hypoxia for 8 and 14 days. Local cardiac ACE expression was studied by immunohistochemistry using a monoclonal antibody to ACE (9B9). RESULTS: In the normal rat heart, ACE expression was confined to vascular endothelium, the valvular endocardium, and localized regions of parietal endocardium. We found that the development of pulmonary hypertension and right ventricular hypertrophy were associated with 2.6- and 3.4-fold increases in membrane-bound right ventricular ACE activity by 8 and 14 days of hypoxia, respectively. Right ventricular ACE activity was positively correlated with the degree of right ventricular hypertrophy (r = 0.83, P < 0.001). In contrast, left ventricular plus septal ACE activity was significantly reduced by approximately 40 and 60% by 8 and 14 days of hypoxia, respectively, compared to controls. In the right ventricle of chronically hypoxic rats, immunohistochemistry demonstrated increased ACE expression in areas of myocardial fibrosis. Interestingly, increased ACE expression was noted in the right ventricular epicardium in chronically hypoxic rats. In the free wall of the left ventricle there was a significant reduction in the number of myocardial capillaries which expressed ACE in chronically hypoxic rats. CONCLUSION: Chronic hypoxia has a differential effect on left and right ventricular ACE activity and that the sites of altered ACE expression are highly localized. We speculate that locally increased right ventricular ACE activity and expression may play a role in the pathogenesis of right ventricular hypertrophy secondary to hypoxic pulmonary hypertension. PMID- 9205555 TI - Circulating transforming growth factor beta 1 and coronary artery disease. AB - OBJECTIVE: Transforming growth factor beta 1 (TGF-beta 1), a multifunctional cytokine, is involved in many physiological and pathological processes and possibly in atherogenesis. METHODS: We explored the association between circulating plasma TGF-beta 1 measured by ELISA and coronary artery disease (CAD) assessed angiographically in 371 Caucasian patients (269 men and 102 women) aged < or = 65 years. RESULTS: While mean +/- s.e.m total TGF-beta 1 was not different among patients with (56.9 +/- 1.5 ng/ml) or without (54.6 +/- 2.8 ng/ml) angiographically demonstrable CAD, naturally active TGF-beta 1 was significantly higher in CAD patients (1.74 +/- 0.18 vs 0.96 +/- 0.17 ng/ml, P < 0.01). Active TGF-beta 1 increased with the number of major coronary arteries with more than 50% luminal obstruction (P < 0.01) and patients with triple vessel disease had twice the level of those with no or mild vessel disease (2.15 +/- 0.46 vs 1.12 +/ 0.14 ng/ml, P < 0.001). We found no relationship between TGF-beta 1 and Lp(a), but TGF-beta 1 was significantly correlated with circulating fibrinogen (r = 0.178, P = 0.005) and fasting glucose (r = 0.177, P = 0.007) levels. CONCLUSIONS: Our study identifies an increase in active TGF-beta 1 levels with both the occurrence and severity of CAD which is independent of standard CAD risk factors. This may reflect a 'double-edged sword' effect of TGF-beta 1 in that it may reduce atherogenesis by inhibiting smooth muscle cell proliferation but, when there is ongoing vessel wall injury, enhance it by promoting excessive extracellular matrix accumulation. The outcome could represent a complex balance between these two competing influences. PMID- 9205556 TI - Basic fibroblast growth factor and heparin influence coronary arteriolar tone by causing endothelium-dependent dilation. AB - OBJECTIVE: The strong angiogenic and mitogenic agents acidic and basic fibroblast growth factors (aFGF and bFGF, respectively) share signalling pathways with known vasodilatory agonists. Therefore, we hypothesized the FGF's produce vasoactive responses. We also proposed that heparin would exert a similar action to FGF, because this proteoglycan not only binds to the FGF receptor, but also facilitates the release of FGF from the cardiac extracellular matrix and promotes its binding to a high-affinity receptor. To test these hypotheses, we examined the vasodilatory reactions of coronary arterioles to aFGF, bFGF, and heparin, and the effects of antagonists to nitric oxide synthase (L-NMMA), prostaglandins (indomethacin), ATP-sensitive potassium (K[ATP]) channels (glibenclamide), FGF and FGF receptors on the vasoactive responses. METHODS: Arterioles (70-110 microns, internal diameter) were dissected from pig hearts and cannulated with micropipettes. Diameter was determined with videomicroscopy in response to bFGF and aFGF in concentrations of 1-100 ng/ml and to heparin (5-200 U/ml). RESULTS: Basic FGF, but not aFGF, caused dose-dependent vasodilation with a maximum of 61 +/- 4%. Relaxation of bFGF was antagonized by pretreatment with L-NMMA, but was not affected by pretreatment with indomethacin or glibenclamide. Heparin caused dose-dependent vasodilation with a maximum of 100 +/- 3% which was partially blocked by either L-NMMA or glibenclamide, but not by indomethacin. Furthermore, the effect of bFGF could be significantly blocked by pretreatment with an FGF receptor antibody as well as with a monoclonal antibody against FGF. Pretreatment with both antibodies significantly inhibited also the effect of heparin. CONCLUSIONS: These results indicate that bFGF and heparin cause vasodilation of coronary arterioles via an increase in NO production and heparin additionally by other mechanisms such as by activating K[ATP] channels. Furthermore, the effect of heparin is partially mediated via FGF and FGF receptors. We therefore speculate that both substances may be involved in the regulation of coronary microvascular tone acting partially through the same signalling mechanisms. PMID- 9205558 TI - The pathogenesis of arterial aneurysms and associated lesions. PMID- 9205557 TI - Expression of phenotype- and proliferation-related genes in rat aortic smooth muscle cells in primary culture. AB - OBJECTIVES: After endothelial injury, smooth muscle cells (SMCs) in the arterial media are modified from a contractile to a sympathetic phenotype. This process includes a prominent structural reorganization and makes the cells able to migrate into the intima, divide, and secrete extracellular matrix components. A similar change occurs in culture and then in vitro system has been established as a useful model in which to study the control of SMC differentiation. The purpose of this study was to analyze the expression of a number of phenotype- and proliferation-related genes in vascular SMCs during the first week in primary culture. METHODS: SMCs were enzymatically isolated from rat aorta and seeded on substrates of fibronectin (an adhesive plasma protein) and laminin-collagen type IV (two major basement membrane proteins) in a serum-free medium or in uncoated dishes in a serum-containing medium. Total RNA was isolated from the cells after different times of culture and analyzed by Northern blotting for expression of specific gene transcripts. In part, expression of the corresponding proteins was also explored by Western blotting and indirect immunofluorescence microscopy. RESULTS: The results indicate that the proto-oncogenes c-fos, c-jun and c-ets-1 were already activated during the isolation of the cells and then continued to be strongly expressed for a few days. Especially in the serum-free groups, there was also early activation of the genes for the matrix metalloproteinases, stromelysin (MMP-3) and type IV collagenase (MMP-2). In parallel, an increased expression of the genes for two extracellular matrix components was observed, with an early rise in osteopontin mRNA and a later rise in collagen type I mRNA. At the end of the test period, the corresponding proteins were deposited around the cells in a fibrillar pattern. Among the matrix receptors investigated, the beta 1 integrin subunit showed a high and persistent expression, whereas the alpha 5 and alpha 1 integrin subunits showed lower and more variable mRNA level. In support of the existence of an autocrine or paracrine platelet-derived growth factor (PDGF) loop, an early rise in expression of the PDGF A-chain gene and a subsequent rise in expression of the PDGF alpha-receptor gene were noted. CONCLUSION: It is proposed that the coordinated shift in gene expression here described to take place in connection with the phenotypic modulation of vascular SMCs in primary culture is part of a predetermined genetic program that normally serves the function to engage the cells in a wound healing response. PMID- 9205559 TI - Natural contamination of spring barley with group A trichothecene mycotoxins in south-eastern Poland. AB - Strains (10705) of microscopic fungi were isolated from spring barley heads in the region of Lublin (south-eastern Poland). Fusarium sporotrichioides Sherb was found in 418 (3.9%) of isolated strains. Group A trichothecene mycotoxins were detected in the collected barley kernels colonized by F. sporotrichioides, with Fusarium head blight symptoms. Among 24 samples analysed, 12 were T-2 toxin positive in a range of contamination from 0.02 to 2.40 micrograms/g (average 0.45), while in five samples HT-2 toxin ranged from 0.01 to 0.37 micrograms/g (average 0.23) and T-2 tetraol was detected in two samples in a range of 0.01 0.21 micrograms/g (average 0.11). Twelve samples were free of detectable amounts of the toxic metabolites analysed. PMID- 9205560 TI - Natural occurrence of deoxynivalenol in wheat, wheat flour and bakery products in Argentina. AB - The objective of this study was to evaluate the natural occurrence of deoxynivalenol (DON) in wheat, wheat flour and different kinds of breads and pastries widely consumed by the population in Argentina. Of 60 wheat samples analysed, 93.3% were contaminated. The average DON contamination level over all samples was 1798 micrograms/kg, and the minimum and maximum values were 100 micrograms/kg and 9250 micrograms/kg, respectively. The wheat flour samples (61 samples) were contaminated with DON at levels ranging from 250 micrograms/kg to 9000 micrograms/kg with an average of 1309 micrograms/kg. The frequency of DON contamination over 42 samples of different bakery products was 92.8%, with levels ranging from 200 micrograms/kg to 2800 micrograms/kg with an average of 464 micrograms/kg. These results suggest a high risk for consumers of wheat products and the need to monitor final products before consumption. PMID- 9205561 TI - Multi-year monitoring of Canadian grains and grain-based foods for trichothecenes and zearalenone. AB - Monitoring of Canadian grain crops and foods by the Health Protection Branch for deoxynivalenol (DON, vomitoxin) has been undertaken every year since 1980, when it was found in Ontario soft wheat for the first time (in the 1979 and 1980 crops). Contamination of this wheat crop has varied, with 22-100% incidences in all but 1 year and up to 0.75 micrograms/g for the annual means of positive samples. The Canadian guideline for DON is 2 micrograms/g in uncleaned soft wheat. Western Canadian hard wheat had < 10% incidence of DON in 7 crop years but 11-43% of samples analysed in 10 other years were positive. Wheat foods, including imports, have shown 9-90% incidences with annual mean levels of 0.07 0.58 micrograms/g in positive samples. Consistently high contamination of Ontario corn has been observed (13-100% annual incidences and annual means of positives 0.16-1.4 micrograms/g). Other trichothecenes, namely nivalenol and HT-2 toxin, have been found infrequently in Canadian grains. New analyses of Canadian and imported beers showed low ng/ml levels of DON. Grains destined for food use and corn foods have been analysed for zearalenone from 1986 to 1993. The most contaminated crop was Ontario; annual mean levels in positive samples ranged from 23 to 215 ng/g. Zearalenone has been detected infrequently in wheat, barley and soybeans (< 75 ng/g). PMID- 9205562 TI - Aflatoxin M1 occurrence in samples of Grana Padano cheese. AB - A total of 223 samples of Grana Padano cheese manufactured in 4 years (1991-94) by dairies in 11 provinces of the Po valley were checked for aflatoxin M1. Grated cheese was extracted with chloroform and the defatted extract was purified by an immunoaffinity column; aflatoxin M1 was determined by HPLC using a fluorescence detector. From the analysis of the data it has emerged that only one sample exceeded the maximum tolerated level in cheese in some European countries (250 ng/kg). Most samples (91%) were in the range 5-100 ng/kg and only 15 (6.7%) in the range 100-250 ng/kg. Notwithstanding a diffuse microcontamination, the situation regarding the AFM1 levels can be considered fairly satisfactory. Mean contamination levels of 1992 and 1994 were significantly higher (P < 0.05) than those of 1993 and 1991. No significant difference was observed among provinces or dairies of origin. PMID- 9205563 TI - Migration from PVC cling films compared with their field of application. AB - Samples of PVC cling films were taken at importers, wholesalers and retail shops, and their overall migration to the alternative food simulant iso-octane was measured, after establishment of a correlation between overall migration to olive oil at 40 degrees C in 10 days and to iso-octane in 2 h. Results of the migration testing were compared with the recommended and/or actual use of the PVC film and the labelling discussed in relation to the relevant EEC directives on food contact plastics. The correct labelling of plasticized PVC film intended for use in retail packaging is important to avoid the risk of significant consumer intakes of the plasticizer di-(2-ethylhexyl) adipate (DEHA) after the film has been used in contact with fatty foodstuffs. Sixty percent of the films declared for use in contact with fatty foods showed too high overall migration compared with the current interpretation of legislation at the time of sampling. In most instances DEHA made up about 80% of the total amount of plastic constituents migrating to iso-octane. Taking into consideration a specific migration limit of 3 mg DEHA/dm2, 77% of the films used for fatty foodstuff analysed were not acceptable. The migration of DEHA to non-fatty foods defined as the food simulant water was at or below 0.1 mg/dm2 in all PVC-films. PMID- 9205564 TI - Food control surveys of polychlorinated dibenzo-p-dioxins and dibenzofurans and intake estimates. AB - Several countries have congener-specific data on polychlorinated dibenzo-p dioxins (PCDD) and dibenzofurans (PCDF) in basic foods as well as calculations of the daily exposure to PCDD/F through food. In many countries the total daily intake is of the same level, close to 2 pg toxic equivalents (TEQ)/kg bw/day. The highest concentrations of chlorinated dioxins and furans have been measured in fish and beef which are also their most important sources, together with milk and dairy products. Compared with adults, the neonates and probably older children have a higher exposure level. If human exposure to chlorinated dioxins and furans is higher than the most common tolerable daily intake (TDI) of 10 pg TEQ/kg bw/day, effective means to control the exposure level are needed. The most important countermeasures are to set limits for emissions of PCDD/F and for concentrations in cow's milk, which is an important source of these compounds for children. PMID- 9205565 TI - Estimation of menthol in Pan Masala samples by a spectrophotometric method. AB - Recently, the Prevention of Food Adulteration Act of India has fixed the level of menthol addition to Pan Masala at 0.1%, therefore good manufacturing practice (GMP) should be adopted so that the samples do not exceed 0.1% menthol (1 mg/g). The estimation of menthol in Pan Masala samples involves steam distillation followed by reaction with p-dimethyl amino benzaldehyde (DMAB) in acidic medium to give a red colour which is read at 550 nm. The sensitivity of this procedure is 75 micrograms menthol per g sample. Using this method, 130 branded and 53 non branded samples of Pan Masala were analysed for menthol content. Almost 25% of branded samples contained less than 1 mg menthol per g while 75% of samples contained 1.1-6.5 mg menthol per g Pan Masala. Non-branded Pan Masala contained 1 mg menthol per g in only 7.6% of samples. However, 92% of samples contained 1.1 6.5 mg menthol per g, suggesting that the addition of menthol is relatively higher in non-branded Pan Masala samples than in branded ones. PMID- 9205566 TI - Reproductive and neurobehavioural effects of lac dye administered in the diet to mice. AB - The natural colour additive, lac dye, was given in the diet to provide levels of 0 (control), 0.15, 0.30, and 0.60%, from 5 weeks of age in the F0 generation to 9 weeks of age in the F1 generation in mice, and selected reproductive and neurobehavioural parameters were measured. There were no adverse effects of lac dye on either litter size or litter weight and sex ratio at birth. The average body weight of offspring during the late lactation period was significantly increased in treatment groups of each sex. In the neurobehavioural parameters, swimming head angle was significantly affected in male offspring during the early lactation period in a dose-related manner, and olfactory orientation was significantly accelerated in female offspring in a dose-related manner. The dose levels of lac dye in the present study produced few adverse effects in reproductive and neurobehavioural parameters in mice. PMID- 9205567 TI - Effect of iron and lactose supplementation of milk on the Maillard reaction and tryptophan content. AB - New liquid UHT milks supplemented with iron (0.9-1.4 mg/100 ml), vitamin C (1-7 mg/100 ml), lactose (2-4 g/100 ml) and linoleic acid (200-400 mg/100 ml), named growth milks, have recently become available to satisfy the specific nutritional needs of children aged 1-3 years. But the iron-vitamin C mixture could activate the lactose-induced Maillard reaction and tryptophan (Trp) oxidation in proteins. We have therefore examined the Amadori product and Trp concentrations of these milks. Forty-two commercial growth milks from five firms were analysed for the Maillard reaction and the soluble protein Trp content and compared with 64 UHT milks. The furosine concentration of total proteins was two to four times higher in 'growth' milks than in standard UHT milks, indicating a proportional loss of available lysine. The Trp fluorescence of undenatured proteins soluble at pH 4.6 was almost three times lower in 'growth' than in standard milks and Trp concentration 36% lower suggesting destruction of this oxidation-sensitive amino acid. The mechanism of Trp destruction remains to be elucidated, and the roles of iron and Amadori products determined. PMID- 9205568 TI - An overview of the safety evaluation of the Thermomyces lanuginosus xylanase enzyme (SP 628) and the Aspergillus aculeatus xylanase enzyme (SP 578). AB - Xylanases SP 628 and SP 578 were produced by submerged fermentation of Aspergillus oryzae, containing a gene code originating from Thermomyces lanuginosus and Aspergillus aculeatus, respectively. Both enzymes were subject to the same series of toxicological tests to document their safety in use. The enzymes are to be applied as processing aids in the baking industry and in wheat starch separation. Neither enzyme was found to be mutagenic in the Salmonella typhimurium reverse mutation assay, nor did they cause chromosomal aberrations in cultured human peripheral lymphocytes. No evidence of inhalation toxicity or skin and eye irritation was found. The enzymes are not regarded as skin-sensitizers, although the Buehler test with guinea-pigs revealed a minor potential. Oral administration up to 10.0 ml/kg bw/day (equivalent to a Total Organic Solids amount of 13.3% for SP 628 and of 11.3% for SP 578) in 13-week rat studies did not show any adverse effect. PMID- 9205569 TI - Analytical quality assurance for the WHO GEMS/Food-EURO programme--results of 1993/94 laboratory proficiency testing. AB - As a means of assessing the performance of European laboratories who contribute analytical data on food contamination to the World Health Organization (WHO) Global Environmental Monitoring Scheme (GEMS), a series of five proficiency testing exercises were carried out during 1993 and 1994. In total 136 laboratories from 21 different countries took part in one or more of the exercises which covered the analysis of trace elements (lead, cadmium and mercury) in milk powder, pesticides (organochlorine, organophosphorus and pyrethroid) in spinach powder, nitrate in spinach powder, aflatoxins in nut-based animal feed and patulin in apple juices. The proficiency testing was carried out according to the ISO/IUPAC/AOAC INTERNATIONAL Harmonized Protocol and laboratories were awarded z-scores signifying their analytical capability based on their reported results for each of the respective exercises. Overall 60% of laboratories were satisfactory for accuracy for trace element analysis, 41% for pesticides, 43% for nitrate, 88% for aflatoxins and 53% for patulin. These results gave an overall poorer performance (68%) than the average for other similar schemes (79%), indicating the need for care in collating data for such programmes as GEMS and the need for remedial measures to assist in improving performance. PMID- 9205570 TI - The fat embolism syndrome. PMID- 9205571 TI - Ipsilateral knee ligament injuries and open tibial diaphyseal fractures: incidence and nature of knee ligament injuries sustained. AB - Fifty patients with isolated open tibial shaft fractures were reviewed to determine the incidence and type of knee ligament injuries sustained. Eighteen patients (36 per cent) had at least one ligament injury in the ipsilateral knee; eight had multiple ligament injuries. Only four patients (22 per cent) were diagnosed as having a ligament injury at the time of initial management and the remaining 14 patients were diagnosed at the time of review for this study. There is a high incidence of ipsilateral knee ligament injuries in open tibial shaft fractures; the knee should be thoroughly examined at the time of initial fracture management. PMID- 9205572 TI - Subtrochanteric fractures of the femur. AB - A consecutive series of 103 patients with a subtrochanteric fracture were prospectively studied. Ten patients were treated non-operatively, whilst the other 93 had operative treatment. The overall fixation failure rate was 12 per cent with a re-operation rate of 6 per cent by 1 year. There were six (8 per cent) failures of fixation for the 74 fractures treated with the sliding hip screw. No method of fracture classification was demonstrated to be of value in predicting either the choice of treatment or the risk of fracture healing complications. Either intramedullary nailing or extramedullary fixation with a dynamic hip screw appear to give the best results for subtrochanteric fractures. PMID- 9205573 TI - Trauma triage: a comparison of CRAMS and TRTS in a UK population. AB - The CRAMS scale and the Triage Revised Trauma Score (TRTS) were compared to assess their potential use as a prehospital method of activating hospital trauma teams. We studied patients from the resuscitation room of Leeds General Infirmary who had enough data recorded to allow calculation of the admission TRTS and CRAMS scale. Patients were defined as having major injury if they died in hospital, were admitted to the ICU or had an Injury Severity Score (ISS) of > 15. Each triage scale was compared by calculating multiple sensitivity/specificity pairs and plotting the results on a receiver operator (ROC) curve. The optimal cut-offs on each scale were compared directly. Ninety-seven (46 per cent) of a total of 213 patients fulfilled the study criteria for major injury. The best cut-off points were a CRAMS of < 9 and a TRTS of < 12. The TRTS was significantly more specific (0.9 versus 0.75) but at a cost of poor sensitivity (0.6 versus 0.69, not significant). The performance of both scales was similar when compared on the ROC curve. CRAMS and the TRTS were unable to identify major injuries in our sample with sensitivity and specificity adequate to support their use as a tool to activate trauma teams in the UK. PMID- 9205574 TI - Vascular injuries associated with limb fractures. AB - The records of 46 patients with vascular injuries of the lower and upper limbs associated with bone fractures and managed in the authors' vascular and orthopaedics divisions were reviewed. All were young men, 26 with blunt and 20 with penetrating vascular injuries; 20 were treated by end to end anastomosis, 12 by vein interposition grafting, six by prosthetic graft, six by vein patches and lateral sutures and two by ligations. The limb salvage rate was 93 per cent. Three patients died (6.5 per cent) of severe associated injuries. Amputations were needed in three patients (6.5 per cent) with popliteal, tibial arteries and vein injuries. An aggressive approach to limb salvage is needed in these extensive injuries. PMID- 9205575 TI - Osteocalcin expression in the groove of Ranvier of the rabbit growth plate. AB - The biology of the growth plates of rabbits of different ages was studied using an immunocytochemical technique which localized osteocalcin. Immunoreactivity was observed in the cells of the groove of Ranvier. Other findings suggest that the groove is a separate structure from the cartilaginous physis and is the terminal end of the diaphysis/periosteum complex. PMID- 9205576 TI - Diaphragmatic injuries. AB - A 6-year series of 26 patients with diaphragmatic injury is presented, 15 with rupture from blunt injuries and 11 after penetrating injuries. All had associated injuries and seven died because of these. The diagnosis may be difficult and was consequently delayed in two patients. Eleven ruptured diaphragms were diagnosed before operation, 14 on the operating table and one at autopsy (dead on arrival). Herniation of abdominal organs was seen in nine of 15 patients after blunt injuries. In most patients repair was via laparotomy using absorbable sutures. It is still essential that the surgeon should be aware of the possibility of the diagnosis and the associated severe injuries. PMID- 9205577 TI - The existence and composition of Trauma Teams in the UK. AB - To determine the existence and composition of Trauma Teams in UK hospitals postal and telephone enquiries were made to 185 Accident & Emergency (A&E) departments comprising all those in the UK with an annual attendance of over 30,000 patients per year. The existence of Trauma Teams was not influenced by the number of A&E attendances or the medical staffing composition in the A&E department. Of the hospitals surveyed, 113 (61 per cent) had either a Trauma Team or an adequate system for getting A&E staff or other specialists rapidly to injured patients. The most common reason (58/69) for not having Trauma Teams was an inability to get doctors to attend to an injured patient promptly. In 58 out of the 69 hospitals the difficulties in getting the appropriate doctors to respond quickly to multiply injured patients was the main reason for not making any arrangements for rapid multidisciplinary evaluation of them. Changing the medical profession's attitude to injury management is a hurdle that will need to be overcome. PMID- 9205578 TI - The triple wire technique for bifacet dislocation of the cervical spine. AB - A prospective study was undertaken to evaluate the triple wire technique in 34 patients who had sustained bifacet cervical spinal dislocations. Skull traction was applied on admission in all patients and the dislocation as gradually reduced under sedation. The operation was undertaken as soon as the general condition of the patient permitted. A triple wire construct utilizing an iliac graft was used in all cases. The patients were mobilized with a soft collar following operation. All grafted areas united and there were no malunions. No patient deteriorated neurologically. PMID- 9205579 TI - Gastric serosal patch in distal pancreatectomy for injury: a neglected technique. AB - Distal pancreatectomy to manage disruption of the body and tail of the pancreas is a well-established surgical procedure. Fistula formation after distal pancreatectomy for injury may be as high as 24 per cent, and its treatment, although non-operative, prolongs hospitalization and increases the patient's discomfort. We describe the gastric serosal patch technique designed to cover the pancreatic stump after distal pancreatectomy in injured patients. Although this procedure has been previously described, it did not receive appropriate acclaim. Our experience suggests that this technique may eliminate fistula formation and other complications, thereby reducing patient discomfort, morbidity and hospital stay. PMID- 9205580 TI - Missed Monteggia fracture dislocation in children. AB - Open reduction of the radial head and reconstruction of the annular ligament has been advocated for the Monteggia fracture dislocation in children who present more than a month after injury. Three patients with an anterior Monteggia lesion were treated by open reduction of the radial head which was held in place by a Kirschner wire passed from the humerus to the radius. No attempt was made either to repair or reconstruct the annular ligament. The patients were aged between 2 and 6 years, the delay between injury and reduction was between 6 and 8 weeks, and the length of follow up was 5 years for two patients and 1 year for the third. All three patients were free of pain, had no deformity and the radial head had not subluxated. All had nearly full flexion at the elbow. The forearm had full supination but restricted pronation. PMID- 9205581 TI - Percutaneous cannulated screw fixation of femoral neck fractures: the three point principle. AB - Out of a total of 183 patients with displaced intracapsular fractures of the femoral neck, 40 were treated with percutaneous cannulated screw fixation according to the 'three point principle'. These patients were reviewed retrospectively, paying special attention to mechanical failure. Thirty-five fractures healed in a good position. Four fractures (10 per cent) showed signs of recurrent instability, resulting in non-union in two and malunion in one. In all four cases of mechanical failure faults in the operative technique or indication could be established. We conclude that percutaneous cannulated screw fixation is a promising method. However, the right indication and a precise operating technique are important. PMID- 9205582 TI - Comparison of intra-articular lignocaine and a suprascapular nerve block for acute anterior shoulder dislocation. AB - We compared the analgesic effects of a suprascapular nerve block with intra articular local anaesthetic in 20 patients presenting with acute anterior glenohumeral dislocations. The intra-articular local anaesthetic technique was a simpler procedure which provided significantly more analgesia for patients. PMID- 9205583 TI - Haemorrhagic hydrops of the gallbladder. PMID- 9205584 TI - Haemarthrosis following thrombolytic therapy for acute myocardial infarction. PMID- 9205585 TI - Obstetric brachial plexus palsy with anterior dislocation of the shoulder. PMID- 9205586 TI - Posterior dislocation of the shoulder with ipsilateral humeral shaft fracture: a very rare injury. AB - A third case of a posterior dislocation of the shoulder with ipsilateral humeral shaft fracture is described. It is recommended that this difficult management problem requires internal fixation of the humeral shaft fracture to allow control of the shoulder. PMID- 9205587 TI - The use of air weapons in attempted suicide. AB - The use of airguns in attempted suicide is uncommon. In such instances, the surface wounds caused by discharged pellets may be inconspicuous or appear deceptively trivial to the medical examiner. Airgun pellets however are easily capable of penetrating the skull or abdominal cavity when fired at the close ranges involved in suicide attempts. The destructive power of these weapons at close range should not be underestimated. We describe three cases of attempted suicide and review the other nine cases reported in the medical literature: of the 12 suicide attempts there were three fatalities. Seventeen out of a total of 19 pellets fired penetrated either the cranial or peritoneal cavity or damaged deep structures. Most of the victims were male. The majority of wounds were right sided. Four of the attempts were extremely determined, involving repeated discharge of the airgun or the use of other means to effect suicide. PMID- 9205588 TI - A technique for removal of fractured locking screws from an intramedullary nail. PMID- 9205589 TI - Preventing guide wire removal during intramedullary reaming. PMID- 9205590 TI - An improved screw design for locked intramedullary nailing. PMID- 9205591 TI - Studies on production, physiochemical and sensory properties of a standard kilishi ingredient mix powder. AB - The quality of kilishi, a sun dried roasted meat product vary considerably due to lack of consistent standardized non meat ingredient portion (spices and condiments) used for its preparation. In this study, an instant standard kilishi ingredient mix powder was developed. The mix powder contained 9.1% moisture, 49.7% protein, 8.3% fat, 3.2% fibre, and 4.4% ash. Particle size of the mix developed was finer than the traditional one. Yield of kilishi increased with increasing concentration of the standard mix powder used for infusion. The highest yield (87%) was obtained at 60% slurry concentration of the mix compared to 59% yield when the traditional paste was used for infusion. Sensory attributes of kilishi produced using 60% slurry concentration of the standard mix was rated better than a commercial kilishi product (P < 0.05). PMID- 9205592 TI - Milk-coagulating enzymes of tuna fish waste as a rennet substitute. AB - Extraction and activation of the tuna zymogen is influenced by temperature, pH, salt concentration and freshness of the stomach tissue. The presence of 25% NaCl in the extraction process markedly enhanced the yield of tuna gastric enzyme. The milk-coagulating time for both tuna protease and rennet, at an incubation temperature of 32 degrees C, was dependent, at similar level, on the pH (5.5-6.4) of the milk as a substrate. Tuna protease was less sensitive to losses of activity than rennet at pH values above 6.4. Both enzymes became unstable beyond pH 7.0 and completely lost their activities at pH 8.0. The purpose of this study was to determine the best conditions for recovery of the fish enzyme and to compare its behavior to that of rennet. PMID- 9205594 TI - Optimization of nitrogen recovery in the enzymatic hydrolysis of dogfish (Squalus acanthias) protein. Composition of the hydrolysates. AB - The recovery of nitrogen in the enzymatic hydrolysis of dogfish shark muscle was optimized by the use of response surface methodology. The optimum values for enzyme/substrate ratio, temperature and pH were found to be 3.7% (w/w), 55.3 degrees C, and 8.3, respectively. The dogfish protein hydrolysate produced under these conditions contained a high crude protein concentration (> 85%), and its high nutritional value was indicated by the presence of all essential amino acids, and by high PER values. These results indicate the potential for dogfish protein hydrolysate to be used in lieu of vegetable proteins as a protein supplement in foods. PMID- 9205593 TI - Effects of coffee consumption on iron, zinc and copper status in nonpregnant and pregnant Sprague-Dawley rats. AB - To investigate the influence of coffee consumption on iron, zinc and copper status, 64 Sprague-Dawley female rats were assigned to four treatment groups: nonpregnant-given coffee, nonpregnant-given water, pregnant-given coffee and pregnant-given water. The coffee groups received a freeze-dried coffee solution (1.1% w/v) as the sole source of liquids during the 18-day study period; all groups were provided a purified diet marginal in iron ad libitum. On day 18 (of gestation in the pregnant rats), rats were intubated with 0.033 MBq59Fe after an 8 h fast and killed 4, 8, 12 or 24 h post-gavage. Fluid intake was significantly lower in the coffee groups. Coffee did not affect food consumption, weight gain, hemoglobin or hematocrit in either pregnant or nonpregnant rats. In nonpregnant rats, the percentage of 59Fe was lower in plasma and higher in the spleen and erythrocytes in the coffee group than in controls; in pregnant rats the coffee group had a higher percentage of 59Fe in the stomach (at 4 h only), blood, spleen and bone than controls. There were no significant effects of coffee on tissue concentrations of Fe, Zn or Cu in the nonpregnant group, but pregnant rats given coffee had high concentrations of Fe in the liver, bone and placenta, and low placenta Zn concentrations compared to controls. In fetuses, there were trends towards lower weight, length and percentage 59Fe, but higher liver Fe, in the coffee group than in the controls. The results suggest that coffee intake leads to (a) increased attempts at erythropoeisis, an indication of iron deficiency, in both nonpregnant and pregnant rats, and (b) impaired placental Fe transport. PMID- 9205595 TI - Kinetics of starch gelatinization during extrusion of tarhana, a traditional turkish wheat flour-yogurt mixture. AB - Tarhana, a traditional Turkish cereal food, was extruded at different extrusion conditions (product temperature: 60-120 degrees C; screw speed: 100-300 rpm; feed rate: 10-20 kg/h). The mean residence time and corresponding degree of starch gelatinization data were used to estimate the order of reaction, gelatinization rate constants and activation energy for starch gelatinization. Results indicate that starch gelatinization exhibited an apparent first-order reaction kinetics with a reaction order of approximately 0.8. Activation energy for gelatinization was calculated as 3325kJ/kg mol using the Arrhenius equation. PMID- 9205596 TI - Functional properties of thermally treated legume flours. AB - Functional properties of four thermally treated decorticated legume flours namely, bengal gram (Cicer arietinum), black gram (Phaseolus f1p4o Roxb.), green gram (Phaseolus aureus Roxb.) and lentils (Lens esculenta) were studied. Samples with moisture levels of 3.2, 3.3, 1.3 and 5.0% for all four were subjected to dry heat treatment in a covered vessel in pressure cooker. (Untreated flours served as controls. Thermal treatment lowered nitrogen solubility profiles of all flours and increased water absorption capacities in bengal gram (146) black gram (451) and lentil (206) over control values of 138, 441 and 180 ml/100 g of flour respectively. Fat absorption capacities decreased in thermally treated bengal gram and black gram (242 and 292) as against 298 and 303 ml/100 g for untreated samples respectively. Foaming capacity also showed a decrease in thermally treated bengal gram and black gram by 28 and 53% respectively over controls. Two deep fat fried Indian products namely, 'Seviya' and 'Chakli' were prepared using two of the legumes. Proximate compositional analysis revealed that products prepared with thermally treated flours absorbed less fat. The sensory scores for appearance, texture, flavour and overall quality obtained by Seviya were 6.04, 6.20, 5.98 and 6.40 for products prepared with untreated flour and 5.74, 5.78, 5.70 and 5.68 for product prepared with treated flour respectively. Chakli prepared with thermally treated flour obtained significantly lower scores of 6.08, 5.2, 5.42, and 5.88 as against 6.78, 6.68, 6.68 and 6.88 obtained by products prepared with untreated flour for similar attributes. PMID- 9205598 TI - Evidence based medicine in otolaryngology. PMID- 9205597 TI - Biochemical contents, their variation and changes in free amino acids during seed germination in Terminalia arjuna. AB - The leaves, twigs, stem and bark of T. arjuna were analysed for their protein, phenol, tannin, nitrate, oxalate in addition to vitamin C, anthocyanin and chlorophyll in the leaves. The variation of some of these parameters in the leaves with season and leaf position was also studied. The time course changes in amino acids and protein during seed germination in T. arjuna, showed initial decrease in protein followed by increase at subsequent stages. The seeds contain high level of serine (21.7%) and glutamic acid (22.6%) the later decreased as the germination progressed. After 30 days seeds showed higher amounts of serine (26.0%), valine (2.8%), proline (10.6%), methionine (3.4%), histidine (5.6%) and lysine (7.4%) while threonine, glutamic acid, tyrosine and arginine were in lower amounts than that of initial stage at 0 day. PMID- 9205599 TI - Giuseppe Gradenigo and his contributions to audiology. AB - Giuseppe Gradenigo (1859-1926) was an important figure in the development of Otology. Within this paper the authors consider his background and his particular contributions to the development of various aspects of audiology. PMID- 9205600 TI - Cartilage-sparing otoplasty: a review with long-term results. AB - Prominent ears are the most frequent congenital deformity in the head and neck region. Anatomy of normal and prominent ears as well as the psychological aspects of prominent ears are reviewed. Two types of surgical technique are described with emphasis on the cartilage-sparing technique. A sound pre-operative analysis, focusing on all parts of the deformity, and surgical techniques which are gradually applied to these deformities should result in pleasing, permanent changes for the vast majority of patients. In our opinion, a combination of cartilage-sparing techniques augmented with cartilage-weakening procedures give predictable long-term results with a natural appearing ear and concomitant few, easily treated complications. PMID- 9205602 TI - Modified tragal cartilage--temporoparietal and deep temporal fascia sandwich graft technique for repair of nasal septal perforations. AB - Fifteen cases of nasal septal perforation were repaired with a tragal cartilage- temporoparietal and deep temporal fascia sandwich technique using a modification of the approach previously described (Hussain and Kay, 1992). Successful closure was achieved in 14 patients (100 per cent) after an observation time of up to two years. The operative technique and advantages of the modified approach are described. PMID- 9205601 TI - Pathogenesis of middle-ear effusion in nasopharyngeal carcinoma: a new perspective. AB - The theory that middle-ear effusion (MEE) associated with nasopharyngeal carcinoma (NPC) is merely the result of tensor veli palatinus destruction is deficient because recent studies have shown that many patients with NPC have MEE but no tensor veli palatinus dysfunction. The present study evaluates the relationship between MEE and Eustachian cartilage erosion by NPC and examines the pathogenesis of NPC-associated MEE from a new perspective. Thirty-five patients with NPC were studied by magnetic resonance scans taken along the lengths of the Eustachian tubes. Twenty-four patients had tumour involvement of both sides of the nasopharynx so that 59 ears were available for study. Eighteen ears had MEE of which 12 had Eustachian cartilage erosion (p < 0.00001), Fischer's Exact Test). In ears with MEE, Eustachian cartilage erosion was frequently but not necessarily associated with tensor veli palatinus destruction. We postulate that altered Eustachian tubal compliance as a result of cartilage erosion by tumour is an important reason why middle-ear effusions develop in patients with NPC. PMID- 9205603 TI - Scleroma of the larynx. AB - Twenty-two cases of rhinoscleroma were studied to determine the involvement of the larynx. The study revealed that scleroma affected the larynx in 40 per cent of cases. In the larynx, scleromatous lesions were found in the subglottic region and the disease was observed in stages i.e., atrophic, granulomatous (proliferative i.e., nodular) and a fibrotic or sclerotic (scarring) stage as found in the nose. The role of laryngological examination is established in the diagnosis of asymptomatic and early laryngeal lesions and in assessing response to initial treatment. Extensive granulomatous lesions were treated by open excision by the laryngofissure approach which was found to be the best method resulting in a quick recovery without any evidence of subglottic stenosis. The disease as a whole is discussed in the light of available literature. PMID- 9205604 TI - Different glycoconjugates in the submucosal glands of the supraglottis and subglottis. Lectin histochemistry study in the hamster. AB - A lectin histochemistry study was performed in the supraglottic and subglottic regions of 10 hamsters. The submucosal glands were observed by light microscopy. The supraglottic submucosal glands presented numerous mucous tubules but on the other hand, the subglottic submucosal glands had serous tubules which finished at the distal portion in serous acini. The results suggest that the distribution of fucosylated-mucin and serum-type glycoproteins between the supra- and subglottic submucosal glands suggest a different viscosity and function of the mucus. PMID- 9205605 TI - Day-case adenoidectomy: how popular and safe in a rural environment? AB - In spite of previously favourable reports on day-case adenoidectomy, there are still worries amongst otolaryngologists that such practice is unsafe, especially in a rural environment. A national survey was therefore carried out which shows that only 41 per cent of respondents perform adenoidectomy routinely as day-case, and even fewer in rural areas. A regional audit on day-case adenoidectomy, covering five hospitals, was conducted in East Anglia. Between 1994 to 1995, 73 day-case adenoidectomies were performed and the outcome was compared to those of 183 in-patient adenoidectomies during the same period. The children in the day case group recovered post-operatively even better than the in-patient group. None of them stayed overnight or required re-admission. There was no increased in post operative consultation to the general practitioner. The parents in the day-case group were mostly in favour of the day-case arrangement (88 per cent). The results suggest that day-case adenoidectomy is safe and popular with parents even in a rural environment. PMID- 9205606 TI - Tracheoesophageal puncture using a flexible gastroscope and a percutaneous endoscopic gastrostomy set. AB - Tracheoesophageal puncture is an established procedure for voice restoration following laryngectomy. A method using a flexible gastroscope and a percutaneous endoscopic gastrostomy (PEG) set is described. This overcomes a number of problems and complications which may be encountered when using a rigid endoscope. PMID- 9205607 TI - The syndrome of inappropriate secretion of antidiuretic hormone (SIADH) as a consequence of neck dissection. AB - The syndrome of inappropriate secretion of antidiuretic hormone (SIADH) can have multiple causes. Surgical neck dissections may have an association with this syndrome and represent the basis for this study. A retrospective review of 50 patients undergoing neck dissections was performed to evaluate for the development of hyponatraemia as a consequence of SIADH. Based on the results of this review, a prospective study of 20 consecutive patients undergoing 22 neck dissections was performed to determine the incidence of SIADH. A control group of 25 consecutive patients undergoing major non-neck dissection surgery was also studied. SIADH developed in nine of 50 patients (18 per cent) of our retrospective group with a high incidence of development in those who had jugular vein ligation (JVL) (22 per cent), pre-operative radiation therapy (25 per cent) or squamous cell cancers (32 per cent). SIADH developed in six patients undergoing 22 neck dissections (27 per cent) in our prospective group. A high incidence was also noted for those with JVL (42 per cent), pre-operative radiation therapy (67 per cent) or squamous cell cancer (40 per cent). No patients developed symptomatic hyponatraemia. No patients in the prospective control group developed SIADH. Neck dissection surgery is associated with a significant risk for the development of SIADH. Factors such as jugular vein ligation (JVL), pre-operative radiotherapy and squamous cell cancer appear to increase this risk. PMID- 9205608 TI - Distant metastases in terminal head and neck cancer patients. AB - With improved control of cancer above the clavicles, distant metastases (DM) are frequently more seen and are becoming a more common cause of morbidity and mortality. The present study defined the incidence of distant metastases in a cohort of terminal head and neck cancer patients (HNCP) and compared it to current reported data. The incidence of distant metastases in relation to the primary tumour was evaluated and their impact on survival was assessed. A retrospective survey of patient charts was made, based on the hospice database and original referring hospital charts. Data of 59 patients admitted to the hospice were evaluated. The incidence and location of locoregional and distant disease were studied and effects on survival analyzed. The overall survival from diagnosis to demise was 42.7 months. Thyroid cancer was seen in 20.3 per cent of cases and squamous cell cancer was seen in 59.3 per cent. Distant metastases were found in 83 per cent and 48.6 per cent of patients respectively. Laryngeal cancer patients had a 54.5 per cent incidence of distant metastases. Locoregional disease was seen in 47 per cent of cases and 35.7 per cent of them had distant metastases while a 64.3 per cent incidence of distant metastases was found in cases without locoregional disease. Mean survival was 47.3 months with distant metastases vs 36.5 months without metastases. The difference was not statistically significant. The incidence of distant metastases in squamous cell cancer in terminal HNCP was 48.6 per cent. This is the highest reported incidence of metastases in a clinical series. Patients without locoregional disease had almost a two-fold incidence of metastases. Survival was not affected by metastases in this series. PMID- 9205610 TI - Luc's abscess--a rare complication of middle-ear infection. AB - A case of subperiosteal temporal abscess of otitic origin is presented. This is an unusual complication of otitis media. The pathogenesis of Luc's abscess is different from other extracranial complications of middle-ear infections in that it is not associated with mastoid infection which results in subperiosteal pus formation. Based on our experience and the reports from the turn of the century, we present the presumptive pathogenesis and clinical features. We contend that these patients run an unexpectedly benign course, and require concomitantly more conservative treatment than other otitic abscesses. PMID- 9205609 TI - Distribution of the solitary adenoma over the parathyroid glands. AB - One hundred and ten patients with primary hyperparathyroidism were studied, in which a normal parathyroid gland was found on the same side as an adenoma (both confirmed by histological examination), and the upper or lower location could clearly be defined during surgery. The distribution of the adenomas over the upper and lower glands was unequal: 61.8 per cent in the superior versus 38.2 per cent in the inferior position. Statistical analysis revealed that this is not a random distribution (p = 0.013). The explanation of this relative predilection is unknown. The finding should not influence the surgical procedure for primary hyperparathyroidism. PMID- 9205611 TI - Cochlear implant extrusion in a child with keratitis, ichthyosis and deafness syndrome. AB - We present a child with keratitis, ichthyosis and deafness (KID) syndrome implanted with a Nucleus device. We discuss the would complications of this child and the steps taken to deal with the problems encountered when the wound failed to heal, followed by the partial extrusion of her implant. Early surgical management involved resuturing the wound but when this failed a rotational flap was required to cover the implant package and allow eventual healing. Despite the wound problems and revision surgery she has a good audiological result. PMID- 9205612 TI - Oroantral fistula: a complication of transantral ligation of the internal maxillary artery for epistaxis. AB - Transantral ligation of the internal maxillary artery (IMAX) is a well-described option for surgical management of posterior epistaxis not controlled by anterior and posterior packing. Advocates for this procedure argue that it reduces the morbidity, length of hospital stay and financial cost associated with prolonged nasal packing. The procedure is carried out through a Caldwell-Luc approach and the IMAX is clipped in the pterygomaxillary fossa. Fashioning of a nasoantral window is optional and its inclusion usually depends on the integrity of the sinus ostium. The commonest complications of transantral IMAX ligation occur when local structures including the inferior orbital and anterior superior alveolar nerves are damaged. The incidence of oroantral fistula following IMAX ligation is very low but those cases reported have been associated with the failure to create a nasoantral drainage window. We report two cases of persistent oroantral fistula complicating transantral internal maxillary artery ligation. No nasoantral window was fashioned in either of these cases. PMID- 9205613 TI - Sphenoid sinus mucocoele presenting with isolated oculomotor nerve palsy. AB - We describe two cases of sphenoid sinus mucocoele. Both presented with isolated oculomotor nerve palsy. Mucocoeles involving only the sphenoid sinus are uncommon. They are probably under-diagnosed as they may be asymptomatic or cause non-specific symptoms. Nasal symptoms occur infrequently but the close relationship of the sphenoid sinus to the orbital apex means that ocular symptoms including cranial nerve palsies are a common presenting feature. Involvement of the third cranial nerve in isolation is rare but has important neurosurgical implications which must be excluded before this symptom is attributed to the sphenoid sinus. PMID- 9205614 TI - Surgical management of bilateral immobile vocal folds and long-term follow-up. AB - Sixty-one cases of bilateral immobile vocal folds were classified as traumatic (52.46 per cent), idiopathic (39.34 per cent) or iatrogenic (8.20 per cent). During follow-up the idiopathic group of patients had a better prognosis (p < 0.05) compared to the traumatic or iatrogenic group. A spontaneous recovery was seen in 58.33 per cent of cases in the idiopathic group, 56.25 per cent in the traumatic group and 40.0 per cent in the iatrogenic group within a period of one year. Patients who failed to show spontaneous recovery were either subjected to arytenoidectomy with fold lateralization, endoscopic fold lateralization or laser cordectomy, showing 70.0 per cent, 66.67 per cent and 80.0 per cent recovery respectively. These cases have been discussed. PMID- 9205616 TI - Acute pulmonary oedema complicating laryngospasm. AB - Pulmonary oedema is an uncommon but important complication of laryngeal spasm which in turn occurs more commonly in ENT practice than in most other surgical specialties. A case is reported and the literature reviewed, with particular reference to the proposed pathophysiological mechanism of this phenomenon. PMID- 9205615 TI - Acquired subglottic cysts in the low birth weight, pre-term infant. AB - Although subglottic cysts have previously been reported as a cause of airway obstruction in the neonate, they have previously been considered to be a relatively rare cause. Cystic narrowing of the subglottis has been associated with endotracheal intubation. With improving survival of pre-term infants the incidence of the condition could be expected to rise. Prior to 1996, only 58 cases had been reported in the literature. We believe that the true incidence of the condition has been considerably under-reported. Over a six-month period our unit diagnosed five cases of compressible cysts in the subglottis in low birth weight, pre-term infants. All patients underwent diagnostic microlaryngobronchoscopy and vaporization of the cysts by CO2 laser. Three children required more than one procedure. In all cases a satisfactory airway was achieved. The pathogenesis, diagnosis and treatment of the condition is discussed. PMID- 9205617 TI - Adenoid cystic carcinoma of the sublingual salivary gland in a 16-year-old female -report of a case and review of the literature. AB - Tumours of the sublingual salivary gland are exceptionally rare. The present case report describes an adenoid cystic carcinoma of the sublingual salivary gland occurring in a 16-year-old girl, in itself an uncommon event. In addition, an interesting feature of the presentation was obstruction of the ipsilateral submandibular gland due to involvement of Wharton's duct. PMID- 9205618 TI - A rare case of Salmonella neck abscess. AB - A case of a deep neck abscess caused by Salmonella enteritidis in a 29-year-old previously undiagnosed diabetic patient is reported. Review of relevant literature has shown 11 cases of Salmonella neck abscess. Predisposing conditions include immunosuppression due to any cause. Salmonella sp. should be included in the differential diagnosis of head and neck abscesses in predisposed individuals and treated accordingly. PMID- 9205620 TI - Horner's syndrome: a rare presentation of cervical sympathetic chain schwannoma. AB - We describe a cervical sympathetic chain schwannoma in a 77-year-old woman who presented with a neck mass and Horner's syndrome. Such schwannomas are rare and this is the first documented case of a Horner's syndrome at presentation. The mass was excised via a cervical approach and her post-operative course was uneventful. The prognosis is excellent, with recurrence being rare. A brief discussion of the pathology, presentation, diagnosis, and treatment of this condition is made in this paper. The relevance of the uncertainty in diagnosis is discussed with the message that a pre-operative Horner's syndrome may guide the surgeon in the care of the patient but we suggest that in all cases proper counselling of the possible neurological consequences of this surgery be conducted. PMID- 9205619 TI - Septal, widely infiltrative and clinically occult adenoid cystic carcinoma of the parotid gland. AB - Most salivary gland tumours present with an obvious mass and are usually diagnosed clinically. We present a case of occult adenoid cystic carcinoma of the parotid which, due to its peculiar septal pattern of growth and complicated clinical setting, defied diagnosis for several years. PMID- 9205621 TI - [Results of hearing preservation in surgery of vestibular schwannoma. Value of combined retrosigmoid and middle fossa approaches]. AB - We report our experience of hearing preservation in acoustic neurinoma surgery, using combined retrosigmoid and middle fossa approaches. Fifty neurinomas operated on between 1987 and 1994 were included in this retrospective study. Hearing preservation surgery was performed for patients with grade II or grade III tumors (mean average tumor diameter: 14.4 mm), presenting with normal or serviceable pre-operative hearing (pure tonal average decrease less than 50 dB, speech discrimination score better than 50%). Isolated middle fossa approach was used in 3 cases, isolated retrosigmoid approach in 2 cases. The 45 other cases were operated on using both routes during the same procedure. Total removal of the tumor with anatomic facial preservation was performed in all cases. No death occurred. The facial function assessed 3 months after surgery was good in 84% of cases (House-Brackmann grades I or II). The mean follow-up was 42 months. Post operative hearing was measurable in 48% of cases and serviceable in 30% of cases. The size of the tumor and the level of preoperative hearing appear to be the most important predictive criteria for successful hearing preservation. PMID- 9205622 TI - [Intracranial meningioma in the elderly. Postoperative mortality, morbidity and quality of life in a series of 39 patients over 70 years of age]. AB - The aim of this study was to assess the current morbidity and mortality in patients over 70 operated for intracranial meningioma. PATIENTS AND METHOD: We report a series of 39 consecutive patients (mean age: 73 y) operated for an intracranial meningioma over a period of 5 years (1990-1994). According to the Karnofski scale (KS), preoperative neurological status was inferior or equal to 70 in 21 patients (53.8%) and superior or equal to 80 in 18 (46.2%). All patients were followed up in order to precisely assess their post-operative condition and a computed tomographic scan (CT scan) was performed during the second semester of 1995 (mean follow-up 29 months). RESULTS: Operative mortality and permanent morbidity were respectively 7.6% and 10.3%. In 77% of this series, the KS score checked at the last follow up was 80 to 100 (good outcome). Poor outcome was defined by death or a postoperative (KS < or = 70, the principal cause being an hemorrhagic infarction. Three factors were predictors of poor outcome: poor preoperative neurological condition (KS < or = 70) (p = 0.07), location of the tumor on the base (p = 0.007), and the duration of surgery > 3 hours (p = 0.06). The logistical regression analysis showed that these three factors were independent. Tumor recurrence occurred in 5 (12.8%) of 39 patients. CONCLUSION: Preoperative KS is a prognosis factor, but a poor preoperative condition is not in itself a sufficient condition contraindicating surgery. The rates of operative mortality of 7.6%, and permanent operative morbidity of 10.3% can be given to patients and their families. PMID- 9205623 TI - [Meningiomas of the sellar diaphragm. Apropos of 4 cases]. AB - Since 1987, we have treated four patients with diaphragma sellae meningioma. Tuberculum sellae meningiomas with intrasellar extension were strictly excluded from this retrospective study. A complete tumor removal was performed in two patients. The two other patients underwent post-operative radiotherapy. According to the Kinjo's classification, the four meningiomas were classified as follows: one type A (supradiaphragmatic-prepituitary), one type B (supradiaphragmatic retropituitary), one type C (subdiaphragmatic), one type not described in this classification characterized by sub and supradiaphragmatic extension. Based on our experience and data in the literature, the clinical and neuroradiological features of the diaphragma sellae meningiomas are reviewed. Diaphragma sellae visualization at MRI, which is not always possible, allows to localize the tumor on a supra- or infra- diaphragmatic position and to decide the optimal surgical approach. The diaphragma sellae is more visible on protonic density, or T2 weighted sequences, but can be located on T1-weighted images. Appropriate surgical approaches are the sub-fronto-pterional route for supradiaphragmatic meningiomas and the transsphenoidal approach for subdiaphragmatic meningiomas. PMID- 9205625 TI - [Neurinomas-neurofibromas]. AB - Among primary nerve sheath tumors, schwannomas and neurinomas are the most common. While the schwannomas (neurilemomas) originate in schwann cells, neurofibromas arise from all the constitutive parts of the nerve. The behavior of each tumor is quite different and only neurofibromas may present malignant transformation, especially when arising in patients with Von Recklinghausen disease (NF1 with multiple neurofibromas). PMID- 9205624 TI - [Primary spinal osteosarcomas]. AB - The rarity of primary osteosarcoma of the spine led us to index the 66 reported cases published in literature. From this analysis a difference was found between spinal osteosarcoma and osteosarcoma of the extremities. Tumors of the spine appear to be two times more frequent in the male population in their thirties. The average period between the beginning of the symptoms and the first consultation is seven months. Back pain is permanent and localized to the affected vertebra. In 80 percent of the cases, neurological symptoms already exist at the stage of the diagnosis. Magnetic resonance imaging (MRI), computed tomography and standard X-ray remain complementary in the morphological analysis of this tumor. All the aspects from the lytic to sclerotic forms are noted, although the lytic form is common. Among spinal osteosarcoma, the lumbar vertebrae are the most frequently affected. Diagnosis can only be established by pathology, even though this may also lead to some errors. In all the reported cases surgery is used, but carcinological methodology is not possible and a complete removal of affected tissue is difficult, with this being achieved in only a quarter of the cases. Radiation therapy, when used, requires doses of 70 Gy to 80 Gy without any certitude of controlling the tumour and with high risks of post-radiation complications. Chemotherapy on its own, despite the use of high dose methotrexate, only has a temporary effect due to partial action on the primary center. Twenty years ago, only twenty percent of all patients suffering from osteosarcoma lived beyond two years, with worse prognosis for spinal osteogenic sarcoma. Today, the therapeutic approach for spinal tumors uses techniques developed in the treatment of osteosarcoma of the extremities, which can now expect more than seventy percent of all patients to live beyond five years. Present day methods recommend a rapid confirmation of the diagnosis, and then a neoadjuvant chemotherapy followed by surgery to remove all the affected area. This strategy allows an evaluation of the tumor chemosensitivity and to adapt the treatment in consequence. The latest results of this treatment on spinal osteosarcoma appear to be encouraging. PMID- 9205626 TI - [Primary spinal osteosarcoma. Apropos of a case]. AB - We report a case of primary osteogenic sarcoma of the third lumbar vertebra, detailing the neuroradiologic and therapeutic aspects. The clinical presentation was limited to low back pain which radiated to the left thigh for 5 months. Lumbosacral spine roentgenograms revealed a sclerotic lesion of the left part of the body of the third lumbar vertebra. Treatment consisted of total vertebrectomy, chemotherapy completed with radiotherapy. Fourteen months after a complex combined treatment no recurrence was observed. A review of the literature highlighted the rarity of this tumor. Usually, patients with vertebral osteogenic sarcoma do poorly. Today, the therapeutic approach for these spinal tumors should use techniques developed in the treatment of osteosarcoma of the extremities because of their encouraging results. PMID- 9205627 TI - Calcitonin versus etidronate for the treatment of postmenopausal osteoporosis: a meta-analysis of published clinical trials. AB - This review examines the evidence on the efficacy of calcitonin and etidronate in the prevention of osteoporosis and osteoporotic fractures. MEDLINE was searched for clinical trials calcitonin or etidronate and reviews of the treatment of postmenopausal osteoporosis. The reference sections of the papers retrieved were again searched for trials on the treatments of interest. Two people independently collected data from the trials that met the inclusion criteria of the study. Weighted means in the change in bone mineral density (BMD) and differences in vertebral fracture rates were computed for calcitonin and etidronate separately. The existence of publication bias was investigated by funnel plots of effect size against sample size. Eighteen clinical trials and calcitonin and six with etidronate were included in the meta-analysis. The pooled change in vertebral BMD at the end of the studies was 1.97 (95% CI 1.77 to 2.17) with calcitonin and 3.20 (95% CI 2.92 to 3.48) with etidronate. Pooled change in proximal femur BMD was 0.32 (95% CI -0.27 to 0.91) with calcitonin and 2.42 (95% CI 2.16 to 2.68) with etidronate. The aggregated number of vertebral fractures prevented by the treatment was 59.2 per 1000 patient-years (95% CI 55.1 to 63.3) for calcitonin and 28.3 (95% CI 26.2 to 30.4) for etidronate. With the available evidence we cannot establish the superiority of either of the two drugs for the treatment of postmenopausal osteoporosis. The clinical trials are particularly lacking in data on hip fracture, the most important consequence of osteoporosis. In this situation consideration of the relative costs of the drugs is prominent. PMID- 9205628 TI - Population-based geographic variations in DXA bone density in Europe: the EVOS Study. European Vertebral Osteoporosis. AB - The purpose of this study was to investigate variations in bone density between 16 European populations, 13 of which were participants in the European Vertebral Osteoporosis Study (EVOS). Men and women aged 50-80 years were recruited randomly from local population registers, stratified in 5-year age bands. The other three centres recruited similarly. Random samples of 20-100% of EVOS subjects were invited for dual-energy X-ray absorptiometry (DXA) densitometry of the lumbar spine and/or proximal femur using Hologic, Lunar or Norland pencil beam machines or, in one centre, a Sopha fan-beam machine. Cross-calibration of the different machines was undertaken using the European Spine Phantom prototype (ESPp). Highly significant differences in mean bone density were demonstrated between centres, giving rise to between centre SDs in bone density that were about a quarter of a population SD. These differences persisted when centres using Hologic machines and centres using Lunar machines were considered separately. The centres were ranked differently according to whether male or female subjects were being considered and according to site of measurement (L2-4, femoral neck or femoral trochanter). As expected, bone mineral density (BMD) had a curvilinear relationship with age, and apparent rates of decrease slowed as age advanced past 50 years in both sexes. In the spine, not only did male BMD usually appear to increase with age, but there was a highly significant difference between centres in the age effect in both sexes, suggesting a variability in the impact of osteoarthritis between centres. Weight was consistently positively associated with BMD, but the effects of height and armspan were less consistent. Logarithmic transformation was needed to normalize the regressions of BMD on the independent variates, and after transformation, all sites except the femoral neck in females showed significant increases in SD with age. Interestingly, the effect of increasing weight was to decrease dispersion in proximal femur measurements in both sexes, further accentuating the tendency in women for low body mass index to be associated with osteoporosis as defined by densitometry. It is concluded that there are major differences between BMD values in European population samples which, with variations in anthropometric variables, have the potential to contribute substantially to variations in rates of osteoporotic fracture risk in Europe. PMID- 9205629 TI - Influence of handedness on calcaneal ultrasound: implications for assessment of osteoporosis and study design. AB - Calcaneal ultrasound has been increasingly studied for its potential in the assessment of osteoporotic fracture risk. The accuracy of such an assessment is, in part, dependent on the reproducibility of the measurement. This study examines the impact of handedness on ultrasound measurements [broadband ultrasound attenuation (BUA) and velocity of sound (VOS)] in the calcaneus. Two hundred and sixty-four subjects (57 men and 297 women) aged 51.1 +/- 13.6 years (mean +/- SD) were studied. For each subject, calcaneal ultrasound measurements were performed on both heels with a McCue CUBA ultrasound densitometer. Right-handed dominance (94.7%) was determined by structured interview. In men, BUA measurements were significantly higher on the dominant side: mean difference 4.1 +/- 1.5 dB/MHz (mean +/- SD; p = 0.009), equivalent to 4.2 +/- 1.5% and more than 4 times the average rate of annual change in BUA. The difference between sides was greater in young (< 50 years) than old men (> 50 years). Among the women, the difference was not statistically significant (0.7 +/- 0.9 dB/MHz; p = 0.4); however, it was significant in younger women (20-30 years) (99 +/- 4 vs 90 +/- 4 dB/MHz, p = 0.01). By contrast VOS did not differ between sides in either men or women irrespective of age. Within-subject standard deviation of BUA was 9.8 dB/MHz for men and 8.6 dB/ MHz for women and the component due to right and left difference was 8.4 dB/MHz for men and 6.9 dB/MHz for women. This variability of BUA between right and left heels could increase the false-positive rate by up to 28% for a cut-off of 2 SD below the mean. These data indicate that variation between left and right heel measurements of BUA is higher than that of random error measurements, particularly in men and younger, presumably more physically active subjects. Although VOS measurements were not side dependent, in the smaller number of studies examining VOS and fracture risk, VOS appears to have a weaker predictive power than BUA. Clinical and epidemiological studies involving calcaneal BUA measurements should standardize the side measured to either the dominant or non-dominant heel, to reduce within-subject variation and increase their power. PMID- 9205630 TI - Volumetric bone mineral density using peripheral quantitative computed tomography in Japanese women. AB - The present study evaluated a commercial device for peripheral quantitative computed tomography (pQCT) and examined the age-related changes in normal Japanese women. The volumetric bone mineral density (vBMD) of the distal radius [integral bone mineral density (BMDI), trabecular bone mineral density (BMDT) and cortical with subcortical bone mineral density (BMDSC)] was measured using pQCT (Norland-Stratec XCT960) in 617 healthy women aged 20-79 years and 75 subjects with osteoporosis aged 60-89 years who exhibited at least one vertebral fracture. The short-term precision errors in vivo (CV, %) were 1.1% for BMDI, 1.1% for BMDT and 1.2% for BMDSC. The correlations between pQCT and dual-energy X-ray absorptiometry measurements (Lunar DPX) of the lumbar spine were r approximately 0.8 (BMDI, BMDT and BMDSC). The maximal mean vBMD values were observed between 20 and 49 years; BMDI, BMDT and BMDSC all showed a linear postmenopausal decline averaging 1.1% per year. The overall decreases in vBMD from the peak values in those 70-79 years were 34%, 32% and 33% in BMDI, BMDT and BMDSC, respectively. The diagnostic sensitivity of osteoporosis was expressed as a T-score. T-scores using pQCT were -3.0 (BMDI), -2.4 (BMDT) and -2.9 (BMDSC). Bone mineral measurement of the distal radius may be useful in the evaluation of age-related bone loss and for the diagnosis of osteoporosis. PMID- 9205631 TI - The effects of standardization and reference values on patient classification for spine and femur dual-energy X-ray absorptiometry. AB - The effect of two methods for standardizing dual-energy X-ray absorptiometry (DXA) measurements on patient classification by the T-score has been determined for a group of over 2000 patients. The methods proposed by the International DXA Standardization Committee and the European Community's COMAC-BME group were used in conjunction with young reference data from the major DXA manufacturers, the COMAC-BME group and the third US National Health and Nutrition Examination Survey (NHANES III). The two standardization techniques produced dissimilar classifications as measured by the kappa statistic (kappa = 0.34-0.90), especially for the femoral neck, with up to 24.3% of patients reclassified from osteopenic to normal and 18.6% reclassified from osteoporotic to osteopenic when the standardization method was changed. Considering the effects of both reference data and standardization techniques together, there was a wide variation of patient classification, with the number of patients classified as osteoporotic varying from 9.6% to 21.1% for the postero-anterior spine L2-4 region and from 2.3% to 27.6% for the femoral neck. The agreement between different classifications ranged widely, from very poor to excellent (kappa = 0.02-0.98). The creation of standardized reference data must be an important priority in order to harmonize patient management using standardized BMD measurements. The choice of standardization technique, however, must be addressed in light of the results presented here. PMID- 9205632 TI - Longitudinal changes in ultrasound parameters of the calcaneus. AB - We examined with a median follow-up of 1.4 years (range 1.0-2.0 years) the rates of change per year in ultrasound parameters of the calcaneus. Speed of sound (SOS), Broadband ultrasound attenuation (BUA) and Stiffness were measured twice in 543 subjects (224 men) participating in the Rotterdam Study. SOS fell by -2.5 m/s per year in both sexes (95% CI -4.0 to -1.1 m/s per year in men and -3.6 to 1.4 m/s per year in women). Stiffness decreased by -0.62 (-1.33 to 0.09) per year in men and -0.66 (-1.24 to -0.08) per year in women. In men the rate of change in SOS and Stiffness tended to increase with age. BUA did not change significantly during follow-up in either sex. The prospectively assessed rates of loss differed considerably from those observed cross-sectionally, especially for SOS in men (cross-sectional -0.7 m/s per year, longitudinal -2.5 m/s per year). There was substantial variation between individuals both in changes per year in SOS and in changes per year in BUA. With a median follow-up time of 1.4 years, approximately 27% of the variation in the rate of change for SOS could be explained by measurement error while for BUA this was approximately 9% and for Stiffness 11%. Only a small percentage of subjects had changes larger than could be accounted for by measurement error (SOS: men 26.8%, women 21.6%; BUA: men 28.5%, women: 38.8%; Stiffness: men 32.6%, women 35.1%). The latter may limit the use of ultrasound measurements as a follow-up tool in individuals rather than in populations. PMID- 9205633 TI - Effects of cyclical etidronate therapy on bone loss in early postmenopausal women who are not undergoing hormonal replacement therapy. AB - This study was carried out to investigate the effectiveness and tolerability of cyclical etidronate therapy in the prevention of bone loss occurring in early postmenopausal women who are not undergoing hormone replacement therapy (HRT). A total of 109 Caucasian women aged 45-60 years were treated with etidronate 400 mg/day or placebo for 14 days followed by calcium supplementation 500 mg/day for 77 days. Ninety-one women completed the 2 years of the study. After 2 years, the estimated difference between the two groups as regards lumbar spine bone mineral density (BMD) was 2.53% (SEM 1.07%; p = 0.01); BMD of the hip and wrist were not significantly different between treatment groups. A clear reduction in bone turnover was obtained as evidenced by a significant decrease in serum alkaline phosphatase level and in urinary N-telopeptide/ creatinine ratio in the etidronate group; the difference between the two groups was -12% +/- 3.2% for serum alkaline phosphatase level (p = 0.019) and -22.9% +/- 13.7% for the urinary N-telopeptide/creatinine ratio (p = 0.047). There was no statistically significant difference between the two groups in terms of the serum osteocalcin levels and urinary hydroxyproline/ creatinine and calcium/creatinine ratios. Etidronate was generally well tolerated and its adverse event profile was similar to that of placebo. The results of this study indicate that cyclic etidronate therapy can prevent trabecular bone loss, with no deleterious effect on cortical bone, in the first 5 years of menopause and that it has a very high safety margin. PMID- 9205635 TI - Osteopenia occurs in a minority of patients with acromegaly and is predominant in the spine. AB - Acromegaly may induce abnormalities in bone metabolism; however, there are limited data related to bone mineral density (BMD) in this condition. To evaluate the effects of an excess of growth hormone/ insulin-like growth fractor I (GH/IGF I) in the skeleton, we measured the BMD in spine and femoral region, total body calcium and body composition in 45 patients (24 females and 21 males) aged 21-77 years (median 43 years) with acromegaly for 11.4 +/- 7.5 years (range 0.5-26 years) using a dual-energy X-ray absorptiometer (Lunar DPX). Thirty-four patients had had hypogonadism for 8.6 +/- 6.5 years (1-24 years). Mean serum GH and IGF-I levels were respectively 159 +/- 183 micrograms/l and 843 +/- 497 micrograms/l. Total body calcium was increased in the acromegalics (males: 1272 +/- 217 g, range 916-1816 g; females: 1041 +/- 223 g, range 739-1609 g) when compared with normal individuals (males: 1115 +/- 144 g, range 856-1398 g; females: 909 +/- 144 g, range 511-1311 g; p = 0.01). The lean body mass was significantly higher in acromegalic patients (p < 0.001) compared with normal individuals. There was a tendency for a lower fat percentage in the acromegalics; however, this difference was not significant. Osteopenia (1 Z-score below the mean) was found in the spine in 20% (n = 9) of the patients, while BMD was decreased in the femoral region in only 8.8% (n = 4). The group with osteopenia had a greater duration of hypogonadism than the normal BMD group (14 +/- 11 years vs 4.4 +/- 4.0 years; p = 0.01). A negative correlation was also found between the duration of hypogonadism and BMD in spine (r = -0.4; p = 0.003) and femoral region (r = -0.37; p = 0.013). The hypogonadal patients had a lower BMD in spine (p < 0.005), but not in other regions analyzed. No correlation was found between duration of hypersomatotropism, GH/IGF-I levels and BMD. We conclude that the majority of patients with acromegaly have preserved BMD despite the presence of hypogonadism. PMID- 9205634 TI - A co-twin study of the effect of calcium supplementation on bone density during adolescence. AB - The effect of calcium supplementation on bone mineral density (BMD) was evaluated in female twin pairs aged 10-17 years with a mean age of 14 years. Forty-two twin pairs (22 monozygotic, 20 dizygotic; (including one monozygotic pair from a set of triplets) completed at least 6 months of the intervention: 37 pairs to 12 months and 28 pairs to 18 months. BMD was measured by dual-energy X-ray absorptiometry (DXA). In a double-blind manner, one twin in each pair was randomly assigned to receive daily a 1000 mg effervescent calcium tablet (Sandocal 1000), and the other a placebo tablet similar in taste and appearance to the calcium supplement but containing no calcium. Compliance (at least 80% tablets consumed), as measured by tablet count, was 85% in the placebo group and 83% in the calcium group over the 18 months of the study, on average increasing dietary calcium to over 1600 mg/day. There was no within-pair difference in the change in height or weight. When the effect of calcium supplementation on BMD was compared with placebo at approximately 6, 12 and 18 months, it was found that there was a 0.015 +/- 0.007 g/ cm2 greater increase in BMD (1.62 +/- 0.84%) at the spine in those on calcium after 18 months. At the end of the first 6 months there was a significant within-pair difference of 1.53 +/- 0.56% at the spine and 1.27 +/- 0.50% at the hip. However, there were no significant differences in the changes in BMD after the initial effect over the first 6 months. Therefore, we found an increase in BMD at the spine with calcium supplementation in females with a mean age of 14 years. The greatest effect was seen in the first 6 months; thereafter the difference was maintained, but there was no accelerated increase in BMD associated with calcium supplementation. The continuance of the intervention until the attainment of peak bone mass and follow-up after cessation of calcium supplementation will be important in clarifying the optimal timing for increased dietary calcium and the sustained, long-term effects of this intervention. PMID- 9205636 TI - DXA longitudinal quality control: a comparison of inbuilt quality assurance, visual inspection, multi-rule Shewhart charts and Cusum analysis. AB - The performance of dual-energy X-ray absorptiometry (DXA) instruments can be monitored using various quality control (QC) procedures. It has not been established which of these is most appropriate. The aim of this study was to determine which of four QC procedures is the best to use for longitudinal monitoring. Eighteen centres with DXA instruments scanned an aluminium spine phantom weekly for up to 16 months, and the bone mineral density data were used for the QC procedures. The methods investigated were the instrument's inbuilt quality assurance (QA) procedure, visual inspection, multi-rule Shewhart control charts, and Cusum analysis using a truncated-V mask. True and false positive fractions (TPF and FPF) of each method were calculated, including those for a range of action levels for the Shewhart charts and dimensions of the Cusum mask. For Shewhart, the action levels giving the most desirable TPF and FPF were whole multiples of the standard deviation (SD). For Cusum, the most desirable mask dimensions were 3.6 SD for the total height of the vertical section and 0.9 SD per data point for the gradient of the wings. Predictive power of each method as a means of fault detection was decided by the number of faults detected out of a total of 8 non-mechanical faults subsequently diagnosed. The inbuilt QA detected 2, visual inspection 7, Shewhart chart 7 and Cusum analysis 7. The FPFs were: visual inspection 0.09, Shewhart 0.04, Cusum 0.08. At these levels of FPF, the average time in days (range) from onset of a fault to detection was 39 (6-82) for visual inspection, 39 (4-116) for Shewhart and 21 (1-49) for Cusum. All three "phantom" methods are excellent for DXA QC, with modified Cusum analysis being the most effective. The inbuilt QA appears of little use on its own for longitudinal QC. PMID- 9205638 TI - Hormone replacement therapy in the prevention and treatment of osteoporosis. PMID- 9205637 TI - Bone mineral density & T-scores. PMID- 9205639 TI - Sex steroids and the cardiovascular system. AB - HRT decreases significantly the risk of cardiovascular disease in postmenopausal women. More and more data indicate that this is true not only for unopposed oestrogen but also for combined oestrogen/progestin therapy. The latter point is of utmost importance since the global treatment strategy for women with an intact uterus includes a progestin. PMID- 9205640 TI - Does estrogen replacement therapy protect against Alzheimer's disease? PMID- 9205641 TI - Sex steroids and malignancies. PMID- 9205642 TI - Promoter- and cell-specific responses to sex steroids. PMID- 9205643 TI - Analysis of the molecular pharmacology of estrogen receptor agonists and antagonists provides insights into the mechanism of action of estrogen in bone. PMID- 9205644 TI - Steroid receptor specificity with reference to the estrogen receptor. PMID- 9205645 TI - Skeletal effects of estrogen analogs. PMID- 9205646 TI - Gonadal hormone substitutes: effects on the cardiovascular system. PMID- 9205647 TI - Tamoxifen: from breast cancer therapy to the design of a postmenopausal prevention maintenance therapy. PMID- 9205648 TI - Gonadal hormones. PMID- 9205649 TI - Gonadal hormone substitutes. PMID- 9205650 TI - Opioid rotation: does it have a role? PMID- 9205651 TI - The challenge of evaluating a child bereavement programme. AB - This paper will discuss the challenge of evaluating the efficacy of child bereavement services. Such services are being developed and it is essential that expansion is based on research and evaluation. A literature review details the limited research which has so far been conducted on such interventions. The paper then addresses four key components regarding child bereavement programme evaluation: preliminary evaluation of the Winston's Wish programme; the feasibility and validity of using an experimental method in the evaluation of child bereavement services; the identification of appropriate measures-what are we really attempting to measure with regard to child bereavement interventions; and the importance of measuring the family dynamics of grief-how can we incorporate the Dual Process Model? PMID- 9205653 TI - Support needs in the last year of life: patient and carer dilemmas. AB - The aim of this study was to identify needs for support and problems in the introduction of support to terminally ill patients and their carers. The design involved semistructured interviews with patients and carers as well as a survey of general practitioners' (GPs) views, and took place in GP practices and homes of patients in Cambridgeshire. The subjects comprised 43 terminally ill patients, 30 carers, 80 GPs and 13 of their GP partners. The main outcome measures were quantitative data about additional help required and qualitative data on reasons for reluctance to seek help. Needs for help with transport, personal care and housework were identified. Carers may also need reassurance from health professionals. The need for outside help may at times conflict with the need to preserve independence, dignity and familiar aspects of life. Sometimes carers may feel that there is need for more help, but that this conflicts with patients' wishes. There may also be reluctance to seek help because of a perceived lack of resources and professionals time. In conclusion, an increase in services is necessary but not sufficient to meet patients' needs fully. Services should be introduced in ways that help patients to preserve independence, dignity and familiar aspects of life. The perception of accessibility to health professionals may need to be improved. Carers' needs should be assessed separately from patients' needs. PMID- 9205652 TI - Family coping and bereavement outcome. AB - Using a three-phase longitudinal design, the bereavement of 115 adult Australian families following the death of a parent from cancer was studied. The cohort comprised 115 spouses and 153 offspring: 670 individual responses were obtained. A range of psychosocial variables was studied through a semistructured interview covering the experience of the deceased's illness, death and funeral, spousal health, family coping, sources of support, use of ritual and completion of estate duties. Bereavement outcome was measured by standardized questionnaires of the intensity of grief (Bereavement Phenomenology Questionnaire), depression (Beck Depression Inventory), psychological distress (Brief Symptom Inventory) and social adjustment (Social Adjustment Scale). Those psychosocial variables found to be significantly correlated with bereavement outcome were entered into best sub-set regression analyses. Family coping was the most consistent correlate of bereavement outcome in these regression analyses, which accounted for up to 38% of the variance in grief, 64% in distress, 53% in depression and 46% in social adjustment. The nature of family functioning is a key aspect of social support in influencing the outcome of bereavement. PMID- 9205654 TI - Contribution of a liaison clinical pharmacist to an inpatient palliative care unit. AB - The impact on patient care of interventions made by a liaison clinical pharmacist visiting a busy inpatient palliative care unit were evaluated using a validated six-point scoring system. Interventions made in 13% of patients could improve patient care, save money or both, but rarely involved the drugs that are commonly used for symptom control in patients with terminal cancer. Advice to rationalize inappropriate drug regimens (53%) was the commonest intervention, followed by warnings about drug interactions (24%) and advice about therapeutic drug monitoring (8%). The interventions were evaluated by the pharmacist, a palliative medicine registrar and two independent doctors, confirming that the pharmacist was valid and accurate in assessing her own work. Although more than 60% of interventions could significantly improve patient care, compliance by medical and nursing staff with advice was only 55%, reflecting possible tensions between palliative and general hospital medicine. This survey emphasizes the role of liaison clinical pharmacists in palliative care, the need for much more critical appraisal of prescribing practices and the utility of ranking pharmacist interventions as a quality assurance and educational tool. In particular, providing palliative care for patients with advanced acquired immunodeficiency syndrome (AIDS) is enhanced when a pharmacist with a specialist knowledge of AIDS therapeutics is available. PMID- 9205655 TI - Drug combinations in syringe drivers: the compatibility and stability of diamorphine with cyclizine and haloperidol. AB - The compatibility and stability of 2B combinations of diamorphine hydrochloride (5-100 mg/ml) with cyclizine lactate (5-50 mg/ml), eight combinations of diamorphine (10-100 mg/ml) with haloperidol (2-4 mg/ml) and eight combinations of all three drugs was assessed after storage in 1 ml polypropylene syringes. Samples were stored for periods up to seven days in the light and at room temperature (22 degrees C). Five combinations of diamorphine with cyclizine precipitated immediately upon preparation. After analysis and determination of t90% values (the time taken for 10% degradation). 16 of the remaining 23 combinations were judged to be compatible (no signs of crystallization or precipitation) and stable (less than 10% loss of potency of either drug) after storage for 24 h. After seven days storage only four remained compatible and stable. The results indicate that ratios of diamorphine to cyclizine of 1:1 are stable at concentrations up to 20 mg/ml. An increase in diamorphine concentration necessitates a reduction in cyclizine to 10 mg/ml, and an increase in cyclizine concentration necessitates a reduction in concentration of diamorphine to 15 mg/ml to maintain stability over 24 h. All the combinations of diamorphine with haloperidol remained compatible and stable for seven days. The addition of haloperidol (2 mg/ml) to the diamorphine and cyclizine combinations had no detrimental effect on their compatibility and stability. A stability curve is included as an easy way for palliative care personnel to avoid potential problems with incompatibilities and reduced stability when using these combinations. Furthermore, to reduce the possibility of precipitation with mixtures containing cyclizine, the use of 0.9% sodium chloride should be avoided. PMID- 9205656 TI - Clinical trial of a mucin-containing oral spray for treatment of xerostomia in hospice patients. AB - Thirty-five hospice patients complaining of dry mouth entered a double-blind, single-phase placebo-controlled trial of a mucin-containing oral spray (Saliva Orthana) for the relief of xerostomia. The sprays were administered ad libitum for two weeks by the patients themselves, with nursing help as necessary. A detailed history and examination were undertaken, together with collection of microbiological specimens, at entry and after seven and 14 days of spray usage, respectively. Thirty-one patients were available for follow-up at seven days and 26 patients after 14 days. Relief of oral dryness during the day was reported by 9/15 patients on Saliva Orthana and 10/16 patients on placebo by day 7, with a similar degree of improvement maintained to day 14. The corresponding figures by day 7 for relief of dryness at night were 8/15 for Saliva Orthana and 8/16 for placebo. There were no statistically significant differences between those on active and those on placebo spray for any of the oral symptoms recorded. Neither spray had any major impact on the oral microflora. However, the majority of patients in both treatment groups wished to continue using a mouth spray at the end of their involvement in the trial. Whilst the data from this study provide no evidence for increased benefit of a mucin-containing spray over a mucin-free placebo among xerostomic hospice patients, it is clear that both sprays provided worthwhile symptomatic relief of oral dryness for many of the participants. PMID- 9205657 TI - Transdermal fentanyl for severe cancer-related pain. AB - A prospective phase II study was conducted to define the analgesic efficacy, acceptability and toxicity of the transdermal therapeutic system (TTS) of fentanyl in Chinese patients with severe cancer-related pain. A total of 14 patients was treated with TTS fentanyl at doses ranging from 25 to 100 micrograms h-1; initial doses were chosen according to their previous opioid requirement. Standard supportive therapy was given as required. A brief pain inventory (using a 10-point scale) was used to assess patients at days 0, 7 and 14. Pain control on day 14 with TTS fentanyl was successful in six patients, with a reduction in the common side-effects of other opioids and improvement in general well-being. Seven patients did not complete the 14-day trial: two developed dizziness and nausea within 3 h of application; and in five, TTS fentanyl was insufficiently flexible to control increasing pain during the first week. TTS fentanyl was effective and well tolerated in 43% of patients. Acute dizziness and nausea within the first few hours after application and the relative inflexibility of dose-adjustment both limited the use of TTS fentanyl. PMID- 9205658 TI - Adjuvant psychological therapy for cancer patients. AB - Adjuvant psychological therapy (APT), a brief, problem-focused, cognitive behavioural treatment for patients with cancer, is described. A previously published randomized trial demonstrated a significant reduction in cancer-related emotional distress. APT is recommended for cancer patients suffering from such distress. PMID- 9205659 TI - Is it cancer or not? AB - Two cases are reviewed in whom a positive bone scan led to a diagnosis of metastatic bone disease and hospice referral. One of these was subsequently shown on magnetic resonance image (MRI) scanning to have benign metabolic bone disease; in the other, a repeat bone scan 18 months later confirmed benign pathology. The potential impact on patient and family in these situations is considerable and the usefulness of MRI in distinguishing benign from malignant bone disease is discussed. PMID- 9205660 TI - Selecting an approach and design in qualitative research. PMID- 9205661 TI - Sedation in catastrophic incidents. PMID- 9205663 TI - Auditing the use of radiological investigations in a day centre. PMID- 9205662 TI - Saliva substitutes or stimulants? PMID- 9205664 TI - Morphine and clonazepam combinations in the Springfusor 30 infusion device. PMID- 9205665 TI - Oxygen and genes in health and disease. PMID- 9205666 TI - Twin studies in medicine--what do they tell us? AB - A well-conducted twin study has the potential to evaluate the relative contribution of genetic and environmental factors to a given disease. Many studies are inconclusive because of problems with methodology. This article reviews the principles underlying the classical twin study and then discusses potential pitfalls. Twin studies in three diseases are evaluated, namely, multiple sclerosis, diabetes, osteoarthritis. Studies in multiple sclerosis are thought to be inconclusive. In type 1 diabetes, genetic factors are very important and according to the single study in females with osteoarthritis, genetic factors are also significant. PMID- 9205667 TI - Snake bites by the jararacucu (Bothrops jararacussu): clinicopathological studies of 29 proven cases in Sao Paulo State, Brazil. AB - The jararacucu, one of the most dreaded snakes of Brazil, southern Bolivia, Paraguay and northeastern Argentina, is a heavily-built pit viper which may grow to a length of 2.2 m. Up to 1000 mg (dry weight) of highly-lethal venom may be milked from its venom glands on a single occasion. It has accounted for 0.8% to 10% of series of snake bites in Sao Paulo State, Brazil. We examined 29 cases of proven jararacucu bites recruited over a 20-year period in two Sao Paulo hospitals. Severe signs of local and systemic envenoming, (local necrosis, shock, spontaneous systemic bleeding, renal failure) were seen only in patients bitten by snakes longer than 50 cm; bites by shorter specimens were more likely to cause incoagulable blood. Fourteen patients developed coagulopathy, six local necrosis (requiring amputation in one) and five local abscesses. Two became shocked and four developed renal failure. Three patients, aged 3, 11 and 65 years, died 18.75, 27.75 and 83 h after being bitten, with respiratory and circulatory failure despite large doses of specific antivenom and intensive-care-unit management. In two patients, autopsies revealed acute renal tubular necrosis, cerebral oedema, haemorrhagic rhabdomyolysis at the site of the bite and disseminated intravascular coagulation. In one survivor with chronic renal failure, renal biopsy showed bilateral cortical necrosis; the patient remains dependent on haemodialysis. Effects of polyspecific Bothrops antivenom were not impressive, and it has been suggested that anti-Bothrops and anti-Crotalus antivenoms should be given in combination. PMID- 9205668 TI - Assessing diagnosis in heart failure: which features are any use? AB - We assessed the value of symptoms, past history, medications and signs in the evaluation of patients who might have heart failure secondary to left ventricular systolic dysfunction. An open-access echocardiography service was set up to help identify patients with left ventricular systolic dysfunction who might benefit from treatment with an angiotensin-converting-enzyme inhibitor. History and examination were recorded for each of these patients. The patients were divided into groups according to whether left ventricular systolic function was preserved or not and whether various clinical features were present or not. Of 259 consecutive patients studied, 41 had impairment of left ventricular systolic function as assessed by echocardiography. Past history of myocardial infarction and displaced apex beat were the best single predictors of left ventricular systolic dysfunction as assessed by echocardiography. The combination of past history of myocardial infarction and displaced apex had the best positive predictive value of all. Patients with such clinical features or combinations of clinical features may not need echocardiography, and where access to this resource is limited, it could be reserved for patients without such diagnostic features. PMID- 9205669 TI - Awareness and control of hypertension and hypercholesterolaemia in France and Northern Ireland. AB - We assessed awareness and control of hypertension and hypercholesterolaemia in a cross-sectional study of 586 men from France and 189 from Northern Ireland, aged 35-55, without known coronary artery disease. Prevalence of hypertension was 28% in France and 31% in Northern Ireland (p < 0.42). In France, 70% of hypertensive subjects were aware of their status, vs. 58% in Northern Ireland (p < 0.10). Overall, 40% of subjects with a history of hypertension were untreated, and only 32% of the French and 12% of the Northern Irish subjects treated for hypertension (diet with/without drugs) were normotensive. The prevalence of hypercholesterolaemia was 46% in France and 48% in Northern Ireland (p < 0.62). In France, 59% of hypercholesterolaemic subjects were aware of their status, vs. only 17% in Northern Ireland (p < 0.0001). In both countries, half of those with a history of hypercholesterolaemia were untreated, and only 47% of the French and 43% of the Northern Irish patients treated for hypercholesterolaemia (diet with/without drugs) were controlled. While awareness of hypertension is comparable in France and Northern Ireland, awareness of hypercholesterolaemia is much lower in the latter. Control of hypertension and hypercholesterolaemia in both countries is poor and should be improved. PMID- 9205670 TI - Anti-PL 4 in patients with systemic lupus erythematosus with severe renal and haematological disease. AB - Anti-PL 4 is an autoantibody which binds to a 150 kDa polypeptide and is found in approximately 1% of SLE sera. The clinical and laboratory features of 16 patients who have had anti-PL 4 detected in their serum are presented. Anti-PL 4 is highly specific for SLE (100%) and identifies a population of patients who typically develop severe renal (75%) and haematological disease (100%). PMID- 9205671 TI - Diabetic nephropathy: an observational study on patients attending a joint diabetes renal clinic. AB - Diabetic nephropathy is a major cause of renal failure, accounting for 20% of patients starting dialysis. In clinical trials, effective blood-pressure control, especially by angiotensin-converting-enzyme inhibitors (ACEIs), retards the rate of progression of renal failure substantially. We examined these effects in clinical practice by surveying patients at a joint diabetes renal clinic at Glasgow Royal Infirmary from 1989 to 1995. We examined the relationship between progression of diabetic nephropathy, mean arterial pressure (MAP), and the use of ACEIs. The average MAP of the whole group of patients fell by 7%, the urine albumin:creatinine ratio fell by 29%, and the rate of progression as measured by the slope of reciprocal of serum creatinine with time (l/mumol/day) was slowed, from -4.59 to -2.76. This is equal to delaying the necessity for dialysis by about 2 years. The joint clinic met its aim and was cost-effective. PMID- 9205672 TI - Acute bone-marrow oedema in cyclosporin-treated renal transplant recipients. AB - Transient musculoskeletal pain may occur in renal transplant patients on cyclosporin (CyA). Of 28 consecutive patients transplanted in this unit between 20 January 1995 and 2 May 1996, eight (two published elsewhere) developed this problem. Before transplantation, three of the patients had received prednisone intermittently or continuously for 15, 5 and 2 years, for asthma, crescentic GN and SLE, respectively. All patients had normal hand radiographs prior to transplantation. Five developed acute rejection following transplantation requiring treatment with methylprednisolone; one also required OKT3 (7 days). Weight-bearing joints of the lower limbs became affected at 3-40 weeks (mean 14) following transplantation. MRI changes (T1-, T2-weighted and STIR images) were consistent with acute bone-marrow oedema. Bone scintigrams showed enhanced tracer uptake in affected joints. A spontaneous complete remission occurred in five patients within 4-16 weeks, and this was supported by serial imaging. The other patient underwent core decompression of the femoral heads with relief of symptoms, but MRI showed bilateral avascular necrosis (AVN) of the femoral heads. MRI proved useful in detecting acute bone-marrow oedema and its possible progression to AVN. The former may be either a distinct entity or a forerunner of AVN. PMID- 9205673 TI - Infections and Wegener's granulomatosis--a cause and effect relationship? AB - The association of infections and autoimmune disease has been noted by various authors. Several mechanisms have been proposed to explain this, with no current consensus. Wegener's granulomatosis (WG) is an autoimmune disease involving predominantly the pulmonary and renal systems, and is associated with a distinct autoantibody-the anti neutrophil cytoplasmic antibody (ANCA). Although no solid evidence implicates infections in the emergence of WG, direct and circumstantial data suggest this relation. We review this evidence and discuss possible underlying mechanisms. We emphasize the relationship between infections and ANCA, and their role in the maintenance of the 'on-going' inflammatory response. PMID- 9205674 TI - Inherited thrombophilias. PMID- 9205675 TI - Resistance to activated protein C and factor V Leiden. AB - Over the last four years, there has been an explosion of knowledge about APCr and factor V Leiden. However, there remain a considerable number of difficult clinical areas in which there are no clear answers. Undoubtedly, factor V Leiden is commonly found in association with venous thromboembolic disease in whatever manifestation, but equally it has an unusually high frequency in the general population. Only a small proportion of those that carry the mutation develop a thrombosis. It is estimated that only 6% of those that carry the mutation will develop a thrombosis over a 30-year period, whilst for antithrombin, Protein C or Protein S deficiency, this figure is nearer 60%. Particular areas of difficulty remain in relation to the use of the combined OCP and in the management of the asymptomatic carrier of the mutation in pregnancy. Although the scientific basis of APCr and factor V Leiden is well established, its natural history remains relatively poorly understood, probably as a consequence of its relative novelty. Despite the plethora of new data that have appeared, there remains much to be learnt about factor V Leiden and the APCr phenotype. PMID- 9205676 TI - IgA nephropathy--a disorder of IgA production? PMID- 9205677 TI - Damage occurs early in systemic vasculitis and is an index of outcome. AB - Because death after acute systemic vasculitis is now uncommon, alternative measures of outcome are required. A significant component of patient morbidity is disease-related damage, which can be quantified by the Vasculitis Damage Index (64 items in 11 organ-based systems). We investigated serially the time-course of damage in 120 patients with systemic vasculitis, to determine the earliest indicators of outcome. High damage scores at 2 years after presentation were characteristic of fatal disease (OR 8.1-12.4). Significant damage occurred within 6 months of presentation, and was a feature of fatal disease. More damage occurred after presentation than after relapse. Lung and multi-system damage were early indicators of poor outcome in severe non-fatal disease. Damage occurs early in systemic vasculitis, and is an indicator of poor outcome. This novel observation, together with evidence of persistent subclinical disease activity and the high frequency of relapse, suggests a need for new treatment strategies. Analogy with the management of acute leukaemia suggests a strategy of early diagnosis and intensive induction of remission, with early escalation of treatment for resistant disease. PMID- 9205678 TI - Controlled trial of pulse versus continuous prednisolone and cyclophosphamide in the treatment of systemic vasculitis. AB - Although cyclophosphamide and prednisolone are effective in treating systemic vasculitis, the optimum treatment regimes and duration of treatment are unknown. We randomized 54 patients aged 15-70 years (median 57.5 years) with systemic vasculitis (classical polyarteritis n = 8, microscopic polyarteritis n = 17, Wegener's granulomatosis n = 29) to treatment with either pulse cyclophosphamide and prednisolone (PCYP) (n = 24) or continuous oral and prednisolone and cyclophosphamide, with the latter followed after a median of 3 months (range 1.5 10 months) by azathioprine (CCAZP) (n = 30). Patients on CCAZP were more likely to develop leucopenia (13/30) than patients on PCYP, (7/24) although the difference was not significant. The numbers of infective episodes during follow up were comparable in the two groups at 1.7/patient for PCYP and 1.66/patient for CCAZP. Overall, 26/30 patients (87%) treated with CCAZP developed treatment related toxicity, as did 17/24 patients (71%) treated with PCYP. After a median follow-up of 40.4 months (range 0.7-64.8), there was no difference in the frequency of deaths (PCYP 5, CCAZP 4), relapses (PCCYP 7, CCAZP 8), treatment failures (PCYP 4, CCAZP 4), improvement in disease activity scores or renal function. Survival at three years was 77% in patients treated with PCYP, and 90% in patients on CCAZP (p = 0.38). There was a tendency towards increased toxicity in patients treated with the continuous regimen. PMID- 9205679 TI - Increased salt retention and hypertension from non-steroidal agents in the elderly. AB - We studied blood pressure and natriuretic responses to acute salt loading, and the effect of non-steroidal anti-inflammatory agents on these responses, in five healthy normotensive women aged 65 to 71 years. Five women aged 25 to 31 years acted as controls. Intravenous saline loading, with and without prior ingestion of ibuprofen, was 15 ml/kg/h for 3 h. Baseline blood pressures were higher in the elderly. Saline infusion without ibuprofen raised systolic blood pressure (SBP) by about 25 mmHg in the older group only. Ibuprofen increased baseline SBP in the elderly (129 +/- 6 vs. 116 +/- 5 mmHg, p < 0.05). Saline loading after ibuprofen again raised blood pressure by about 25 mmHg in the elderly only. The elderly group showed markedly increased sodium excretion during saline loading, but this was reduced by ibuprofen. Ibuprofen had no effect on SBP or sodium excretion in controls. Ageing appears to increase susceptibility to salt retention and hypertension from non-steroidal anti-inflammatory agents. PMID- 9205681 TI - The national breast screening service: is it economically efficient? AB - Currently the UK national breast screening programme only offers routine screening to women aged between 50 and 64. Whilst there are good clinical and economic reasons for not screening younger women, there is no compelling argument for not extending routine screening to older women. In this paper, we show that by diverting screening resources to older women, where cancer is more prevalent, more lives and life-years can be saved for no extra cost. Therefore, the current breast screening programme may be inefficient, and offering screening to older women should be given serious consideration. PMID- 9205680 TI - Hepatitis B and C in doctors and dentists in Nigeria. AB - We surveyed a random sample (n = 75) of doctors and dentists at University College Hospital, Ibadan, Nigeria. They were offered anonymous testing for hepatitis B surface antigen (HBsAg), hepatitis Be antigen (HBeAG), antibodies to hepatitis B core antigen (anti-HBc) and to hepatitis C virus (anti-HCV), by enzyme immunoassay. The results suggest a high prevalence of hepatitis B virus (HBV) with a high potential of transmissibility, as well as a high prevalence of HCV infection. The majority of the doctors and dentists use universal precaution for protection against viral hepatitis on < 50% of the occasions when they carry out procedures on their patients. Infection with HBV was associated with type of specialty (surgeons, dentists) and lack of HBV vaccination (p < 0.05). After logistic regression, these factors were independently associated with HBV infection (p < 0.05). Sixty (80%) had not received prior HBV vaccination. Unvaccinated personnel were more likely to be surgeons, dentists, < 37 years of age, and have fewer years of professional activity (p < 0.05). After logistic regression, only fewer years of professional activity remained independently associated with lack of vaccination (p < 0.05). To reduce the occupational exposure of HBV, universal precautions must be rigorously adhered to when the doctors and dentists carry out procedures on their patients, and all health-care workers should be vaccinated with HBV vaccine and the HCV vaccine, when it becomes available. PMID- 9205682 TI - How important is family history in ischaemic heart disease? AB - A positive family history is an established risk factor for ischaemic heart disease, but the size of the contribution relative to classical risks is open to debate. The literature suggests that inherited factors are important in the development of premature ischaemic heart disease, but decline in importance with age. A polymorphism in the angiotensin-converting-enzyme gene was the first new genetic factor thought to contribute independently and significantly to cardiovascular risk. However, more recent large prospective studies have indicated that its contribution is smaller than was originally thought. Interventions should continue to be targeted at the reduction of important environmental factors, such as smoking cigarettes. PMID- 9205683 TI - Treating Wegener's granulomatosis. PMID- 9205684 TI - [Pulmonary echography: a method of the future in emergency medicine and resuscitation]. PMID- 9205685 TI - [Ion transports across the respiratory epithelium]. AB - The electrolytes and water transport across the respiratory epithelium have aroused particular interest for 20 years ago, with regard to the essential role they play in the regulation of the bronchial mucus composition and volume. The development of new in vitro and in vivo, techniques, allow better knowledge of these transport systems and their regulation. Transport involves two main ionic movements: chloride secretion and sodium absorption by the epithelial cells, associated with parallel water movements. In cystic fibrosis, the modification of bronchial mucus results from a defective protein-kinase dependent regulation of chloride secretion. This defect blocks water and chloride secretion by the respiratory epithelium causing dehydration of the mucus. PMID- 9205686 TI - [Pulmonary complications of human immunodeficiency virus infection in sub-Saharan Africa]. AB - Based on a selection of articles published in the literature and reports from international AIDS conferences, we present the main pulmonary complications of HIV-infection observed in sub-Saharan Africa. The different clinical studies demonstrate the predominance of infectious complications, mainly tuberculosis (29 to 44%) and bacterial pneumonia (21 to 35%). The frequency of Pneumocystis carinii pneumonia remains low (5 to 19%). Other complications (mycobacterial infection, cytomegalovirus, toxoplasmosis, cryptococcus, aspergillosis, interstitial lymphoid pneumonia, Kaposi sarcoma) are less frequent. The autopsy studies report similar results and mention the predominance of tuberculosis and pneumonia due to common germs as well as the low frequency of pneumocystosis. This analysis of work conducted in sub-Saharan Africa clearly indicate that tuberculosis remains the leading cause of morbidity and mortality in HIV-infected patients. PMID- 9205687 TI - [Effects of leukotrienes on bronchial secretions]. AB - Leukotrienes have multiple effects on the bronchi (inflammation, bronchoconstriction, increased vascular permeability...). They are also directly involved in regulating the composition of bronchial secretions. In vitro studies in animals and in man have demonstrated that sulphidopeptide leukotrienes stimulate active secretion of chloride by airway epithelial cells, an effect that should favor hydration of bronchial secretions. In addition, sulphidopeptide leukotrienes are powerful stimulants for mucin glycoprotein secretion. This effect is limited, at least in vitro, by inhibitors of the lipoxygenase pathway. These results underline the need for clinical studies to better define the role of leukotrienes in bronchial hypersecretion and to evaluate the effect of anti leukotriene agents. PMID- 9205688 TI - [Aerosol therapy and cystic fibrosis: a national survey]. AB - Aerosol therapy in cystic fibrosis is indicated in order to administer active agents directly into the diseased organ. The practical application of this technique often remains a question of personal experience rather than rigorous and validated schemes. In order to determine the experience of those using this technique, a questionnaire was prepared by the health care centers certified by the French association against cystic fibrosis (65 centers). There were 53 responses (82%) covering 3400 patients. Two-thirds of these patients (2250) used nebulizers. Drugs used by order of frequency were: rhDNase (52 centers), antibiotics (51), conventional fluidifying agents (17), isotonic saline solution (14). Bronchodilators were used as sprays or inhalation chambers. Antibiotics most frequently prescribed were: colimycin (51 centers, doses 500.000 to 3 MIU per aerosol), tobramycin (44 centers, 25 to 600 mg per aerosol), amikacine (7 centers, 150 mg to 1.5 g per aerosol). Indications were mainly chronic colonization with Pseudomonas aeruginosa (39/53 centers) and episodes of bronchial superinfection with Pseudomonas aeruginosa (18/53 centers). Equipment maintenance appeared as the main problem in most centers (daily or more cleaning with antiseptic solutions in 46/53 centers). The differences in indications and dosages prescribed justify further prospective randomized studies to optimize treatment. PMID- 9205689 TI - [Bilateral perihilar alveolar opacities with major changes in health status]. PMID- 9205690 TI - [Left crural monoplegia following pulmonary embolism]. PMID- 9205691 TI - [Oral desensitization to antitubercular agents]. AB - We report a case of hypersensitivity reaction to majors antituberculosis drugs. We have successfully performed oral desensitization to "Rifater *". We present our protocol that we compare to those proposed in the literature. PMID- 9205692 TI - [Prevention multi-resistant bacterial infections in resuscitation (excluding modalities of optimization of antibiotic therapy). 16th Consensus Conference on resuscitation and emergency medicine. 1996 November 21 Thursday, Scientific Information Center of ARC, Villejuif]. PMID- 9205693 TI - [Community-acquired pneumonia of atypical presentation, role of macrolides]. PMID- 9205694 TI - [Genetics of dominant autosomal forms of Alzheimer disease: 3 genes and one phenotype. Groupe de Recherche Francais sur la Maladie d'Alzheimer]. PMID- 9205695 TI - [Mechanism of hypometria caused by pontocerebellar infarction]. AB - By contrast to cerebellar hypermetria, the pathophysiology of cerebellar hypometria associated with a focal lesion is poorly understood. We describe a patient presenting an ischemic stroke involving the left lateral pons and middle cerebellar peduncle and exhibiting severely hypometric ballistic movements of the left wrist. Hypometria was associated with two distinct abnormalities of the triphasic electromyographic (EMG) pattern: a prolongation of the duration of the antagonist EMG activity and a reduced intensity of the agonist EMG activity. The first mechanism has been described previously in patients with a cerebellar disease and exhibiting hypometria, but not the second. The concomitant presence of these two elementary mechanisms contributed to the severity of hypometria in our patient. Moreover, this case illustrates that the integrity of the crossed pontocerebellar fibers passing through the middle cerebellar peduncle is necessary to generate an adequate agonist EMG activity. PMID- 9205696 TI - [Dystonia and tremor in bilateral lesion of the posterior mesencephalon and the vermis]. AB - The aim of this study is to report the association of diffuse dystonia and tremor in a bilateral and extended lesion of the posterior mesencephalon. After surgery on a meningioma of the upper part of the fourth ventricle, this patient presented with facial dystonia, predominating on orbicularis muscles and peribuccal area, and limb dystonia, with tonic extension of fingers and first toes. The tremor was associated with a rhythmic and most often alternate agonist-antagonist muscular activation, whose frequency varied from 3 to 7 Hz. These disorders were increased by the standing position, voluntary movement, somatosensory stimulations, stress or emotion. Pyramidal and somatosensory tracts were spared. Therapeutic trials showed that both the dystonia and tremor were improved by subcutaneous injection of apomorphine, the dystonia by trihexyphenidyle, and the tremor by carbamazepine and propranolol, but not by levodopa and benserazide. The cerebral blood flow study using HMPAO showed a relatively important activity on the cerebellum, which could play a role in the onset of these disorders. PMID- 9205697 TI - [Static contrast sensitivity in idiopathic Parkinson disease]. AB - During Parkinson's disease static contrast sensitivity (C.S.) abnormalities are linked to an impairment of the sensitive visual function. C.S. was tested in twelve parkinsonians and 12 controls without neurological and/or ophthalmological pathology, using a Colored Stripes Electronic Generator (GEPCO). Results for parkinsons showed a general deficiency over the spectrum of spatial frequencies, which was statistically significant and particularly pronounced for intermediate frequencies. This study was repeated for three patients: it showed threshold deterioration for two of them, correlated by evolution in the disease, and an improvement for the third patient after introduction of dopatherapy. C.S. is subjected to dopaminergic control. Among parkinsonians. C.S. deterioration may result in an operating failure of both the visual cortex and retina, and is improved by dopatherapy. The Static Gepco contrast sensitivity test is easy to reproduce and can be used easily to monitor the sensory visual defect in parkinsonian patients under treatment. PMID- 9205698 TI - [Bilateral intra-axial involvement of the common oculomotor nerve (III): 2 cases]. AB - Bilateral palsy of the common ocular motornerve (III) was observed in two patients with an intra-axial lesion due ti hemorrhage in one and ischemia in the other. The lesions involved the cerebral peduncle in the periaqueductal region and the nuclear complex of the III in the first case. Bilateral infarct of the thalamus was seen in the second. Clinical manifestations were transitory except for the oculomotor impairment. In the first patient, oculomotricity was dissociated as intrinsic mortricity was spared. These exceptional cases demonstrate a syndrome with unique oculomotor expression resulting from intra axial oculomotor lesions. Prognosis varies and is related to the ischemic or hemorrhagic nature of the causal lesion and its localization. PMID- 9205699 TI - [Acute Weston Hurst necrotizing hemorrhagic leukoencephalitis]. AB - The clinical and pathological findings of a 43-year-old woman, diagnosed as having acute hemorrhagic leukoencephalitis at postmortem examination, are presented. The acute hemorrhagic leukoencephalitis affects mainly young adults and is the most fulminant from of demyelinating disease. It is frequently preceded by a respiratory infection. Diagnosis is facilitated by CT scanning and MRI, which reveal the massive lesion in the cerebral white matter. Many cases terminate fatally in 2 or 4 days, but in others survival is longer. The pathological findings are distinctive. PMID- 9205700 TI - [Familial cerebral cavernous angiomatosis]. AB - Two patient with familial cavernous angiomatosis presenting with long lasting variable epilepsy with a poor therapeutic response and variable neurologic impairments are presented here. One of the numerous cavernous angiomas was resected in one case. This last patient remains asymptomatic. Familial cerebral cavernous angiomas are often numerous and disseminated in the brain, therefore clinical manifestations are very polymorphous. Moreover the course of these lesions is variable. Therefore MRI should be performed to every patient presenting with poorly understood neurological symptoms, focal or generalized epileptic seizures or absence in order to look for potentially imputable brain lesions. A reliable genetic marker might be helpful for diagnosis of this disease with a variable penetrance and autosomal dominant inheritance. Then a neurosurgical treatment should be carefully discussed if lesions are accessible and medications are poorly efficient with recurrent neurologic impairments or epilepsy. PMID- 9205701 TI - [Cerebral thrombophlebitis, sural phlebitis, pulmonary embolisms and protein S deficiencies]. AB - The authors report about a patient with thrombosis of the transverse and sigmoid sinus who also suffered from multiple pulmonary embolism and deep leg vein thrombosis. The etiologic factor was a deficiency of the free (unbounded) and total protein S. The cerebral sinus thrombosis was diagnosed by MRI and angio MRI. The first 4 weeks the patient was treated with heparin and later with phen procoumon. The plasma protein S serves as a cofactor for protein C and plays an important role in the anticoagulation. Deficiencies of these proteins are either hereditary with an autosomal dominant trait or acquired in patients with severe hepatic diseases and coagulation disorders. PMID- 9205702 TI - [Pure motor hemiplegia with ipsilateral lingual palsy caused by pontine infarction]. AB - A 36 year old diabetic man developed a pure motor hemiplegia (PMH) associated with an ipsilateral lingual palsy. Magnetic resonance imaging revealed a pontine infarct. Lingual palsies have never been reported in patients with PMH so far, but may be associated with other lacunar syndromes such as the "dysarthria-clumsy hand syndrome". This observation supports the hypothesis that corticohypoglossal pathways may have bilateral and assymetrical projections. PMID- 9205703 TI - [Homage to Yvon Lamour (1948-1996)]. PMID- 9205704 TI - Similar electrophysiological correlates of texture segregation induced by luminance, orientation, motion and stereo. AB - Certain local features induce preattentive texture segregation. Recently, components in the visual evoked potential (VEP) associated with preattentive texture segregation (tsVEPs) have been demonstrated. To assess the similarity and dissimilarity of visual processing across visual dimensions, we compared VEPs and tsVEPs in texture segregation by luminance, orientation, motion and stereo disparity. We found tsVEPs across these four visual dimensions to be remarkably similar when compared to the "low-level" VEPs. The tsVEPs were always negative; their implicit time, peak latency and amplitude were (in msec/msec/microV): 91/234/-5.7, luminance; 84/257/-3.9, orientation; 80/295/-8.3, motion; and 95/310/-5.0 for stereo. The cross-correlation function, as a quantitative measure for similarity, on average was higher for the tsVEPs by a factor of 4.2 as compared to the low-level VEPs (P < 0.0001). The results suggest (1) that the tsVEPs represent activity of neural mechanisms that have generalised to some degree across visual dimensions; and (2) that these hypothetical generalisation mechanisms might exist already in the primary visual cortex. PMID- 9205705 TI - Image segmentation enhances discrimination of motion in visual noise. AB - The primate visual system uses form cues-such as hue, contrast polarity, luminance, and texture-to segment complex retinal images into the constituent objects of the visual scene. We investigated whether segmentation of dynamic images on the basis of hue, luminance contrast polarity, or luminance contrast amplitude aids discrimination of motion direction. Human subjects viewed dynamic displays of randomly positioned dots, in which a variable proportion of the dots moved in the same direction at the same speed ("signal" dots) while the remaining dots were randomly displaced ("noise" dots). In agreement with previous reports, we observed a reliable relationship between the strength of the motion signal and subjects' ability to discriminate motion direction, enabling the measurement of thresholds for direction discrimination. When signal dots had a different luminance contrast amplitude than noise dots, direction discrimination performance was directly related to the relative contrast of the signal dots, demonstrating the importance of matching the perceived contrast amplitude of signal and noise tokens when testing the effects of segmentation by other cues. When Michelson luminance contrast was matched, distinguishing signal from noise dots by hue or by luminance contrast polarity strongly improved direction discrimination, lowering thresholds by an average factor of five. These results reveal a strong influence of form cues on motion processing in the human visual system, and suggest that segmentation on the basis of form cues occurs prior to motion processing. PMID- 9205706 TI - Retention of local information in generation of subjective contours. AB - Temporal integration characteristics of subjective contour perception was investigated, using the sequential presentation of two pairs of disks with a sector removed. In the first experiment, by matching the contrast of a "real" stimulus, the perceived contrast of the subjective contours was measured as a function of the stimulus onset asynchrony (SOA) between the two pairs of the disks. With increasing SOA, the perceived contrast decreased gradually and levelled off at the SOA of ca 372 msec (1 SD = 119 msec). In the second experiment, the perceived contrast of the inducing disks was measured as a function of SOA using the matching method. The time limit of the additive effect for the contrast perception of the inducing disks was much shorter than that for subjective contours; the critical SOA was ca 65 msec (1 SD = 36 msec). The remarkable difference of the integration time was explained by a hierarchical process; the local spatial filtering, the retention of local information, and the completion of gaps by multiplicative or AND operation. PMID- 9205707 TI - Light source dependence in shape from shading. AB - We investigated shape constancy in human shape from shading under variations of illuminant direction using a local attitude probe in conjunction with a perturbation analysis. Stimuli were computer generated and depicted ellipsoids in a structured setting. Even with these simple shapes subjects settings were systematically biased in the illuminant direction and were consistent with a regression to image luminance gradients. These biases were reduced for high albedo scenes where interreflections make image illuminance more dependent on scene geometry. Adding texture to the surface reduced but did not eliminate this bias. These results suggest that we can expect little constancy in shape from shading under variations of illuminant direction without constraints from other cues. PMID- 9205708 TI - Contrast detection and orientation discrimination thresholds associated with meridional amblyopia. AB - Humans who have astigmatism resulting in meridional amblyopia exhibit deficits in performing visual tasks at or near detection thresholds. However, there is mounting evidence supporting the idea that performance at threshold may not reliably predict visual capabilities at supra-threshold levels of stimulation. In this study the threshold and supra-threshold performance of six meridional amblyopes were compared. A difference in the pattern of oblique effects was observed between contrast detection thresholds and supra-threshold orientation discriminations. This suggests there exists an independence between populations of neurons subserving contrast detection and the discrimination of visual stimuli. Meridional amblyopia may primarily involve a degradation in those mechanisms subserving visual contrast detection. Populations of cells subserving supra-threshold abilities such as orientation discrimination remain relatively unaffected in humans exhibiting meridional amblyopia. PMID- 9205709 TI - Nonlinear preprocessing in short-range motion. AB - The phenomenon of non-Fourier motion (visually perceived motion that cannot be explained simply on the basis of the autocorrelation structure of the visual stimulus) is well recognized, and is generally considered to be due to nonlinear preprocessing of the visual stimulus prior to a stage of standard motion analysis. We devised a sequence of novel visual stimuli in which the availability of a motion stimulus depends on the nature of the nonlinear preprocessing: an nth order stimulus Pn will generate a perception of motion if it is preprocessed by a nonlinearity of polynomial order n or greater, but not if preprocessed by a nonlinearity of polynomial order less than n. We found that unambiguous motion direction was perceived for P2, P3, and P4, but not for higher-order stimuli, and we measured the contrast thresholds for direction discrimination with superimposed noise. We found that an asymmetric compressive nonlinearity can, in a unified fashion, account for these results, while a purely quadratic nonlinearity or a rectification of the form T(p) = magnitude of p cannot. We compared velocity discrimination judgements for second-order non-Fourier stimuli (P2) with standard drifting gratings. Although velocity comparisons were veridical, uncertainties were greater for the non-Fourier stimuli. This could be reproduced by substituting a Fourier grating with superimposed noise for the non Fourier grating. These findings are consistent with a single pathway which processes both Fourier and non-Fourier short-range motion, and are discussed in the context of other investigations which have been interpreted as demonstrating separate pathways. PMID- 9205710 TI - Motion minima for different directions in color space. AB - We have used the minimum-motion stimulus of Cavanagh, MacLeod & Anstis [(1987) Journal of the Optical Society of America A, 4, 1428-1438] to examine how signals along different directions in color space interact in motion perception. Stimuli were pairs of counterphasing gratings combined 90 deg out of phase in both space and time and modulated along different color-luminance axes. The axis for one of the gratings was fixed, while the axis for the second was varied so as to null perceived motion in the stimulus. The motion nulls show that observers are sensitive to motion signals carried by each of the cardinal directions of color space [an achromatic axis and L-M and S-(L+M) chromatic axes], but that signals along different cardinal axes are not combined to yield a net direction of motion. Pairing an achromatic and chromatic grating resulted in a motion null regardless of the relative or overall contrast of the two gratings, while the null directions for intermediate axes shifted depending on contrast. This result points to the special status of the luminance and chromatic axes. However, our results do not reveal a special pair of axes within the equiluminant plane. When contrasts along the cardinal axes are scaled for equal multiples of their respective detection thresholds, the L-M and S chromatic contrasts contribute roughly equally to the perceived motion, but are many times weaker than luminance contrast. Moreover, sensitivity to luminance motion is little affected by the presence of chromatic contrast, whereas sensitivity to chromatic motion is strongly masked by either luminance or chromatic contrast. These asymmetric interactions suggest that the motion of the luminance and chromatic components is encoded in qualitatively different ways. PMID- 9205711 TI - The effect of attentional spread on spatial resolution. AB - The effects of attentional spread were studied by having subjects detect a luminance increment along a row of evenly spaced dots. The increment could occur for the central, fixated dot (Narrow Attention) or for either the fixation dot or one of the four dots to its left or right (Broad Attention). Narrow Attention enhanced the detection of luminance increments for the fixated dot, and also enhanced spatial resolution near the fixation dot for judgments of vernier alignment and separation. This indicated that the sensitivity of small spatial filters in the fovea was increased more by narrowly focused than broadly spread attention. Effects of attentional spread on spatial resolution were not obtained for judgments of the separation between two peripherally located targets, perhaps because of their dependence on eccentricity (position) rather than separation. PMID- 9205712 TI - Involvement of NMDA in a plasticity phenomenon observed in the adult frog monocular optokinetic nystagmus. AB - The frog horizontal monocular optokinetic nystagmus (H-OKN) is asymmetrical, the reflex being evoked by a temporal-nasal (T-N) component, but not by a nasal temporal (N-T) component. Coil recordings showed that, in adult animals, 8 days of monocular deprivation (by unilateral eyelid suture) provoked the appearance of a N-T component, the H-OKN becoming symmetrical, reacting for both directions of stimulation. This delay was shortened to 2 days following two successive unilateral pretectal administrations of NMDA or of LY 285 265, an NMDA agonist, the first 2 days of eyelid suture. The same results were obtained when chronic microinjections of NMDA or LY 285 265 were achieved, the frogs being maintained in total darkness during the week of eyelid suture. These data indicate that the plasticity phenomenon evidenced in the monocular frog H-OKN depends on the activation of the NMDA receptors of one pretectum. This activation was obtained either by a monocular light stimulation of 8 days duration, or by unilateral administration of drugs activating the NMDA glutamatergic pretectal system. In this last case, the light stimulation was no longer necessary. PMID- 9205713 TI - The development of accommodation. AB - We investigated the roles that blur, proximity and vergence cues play in the development of accommodation. Accommodative responses to targets incorporating one or more of these cues were measured for four adults and eight infants at 1.5 and 3 months of age using eccentric photorefraction. Adults showed accurate accommodation to blur cues, and variable accommodation with proximity cues alone. Some infants at both ages showed fixed accommodative responses to all stimulus conditions. Others responded consistently in the correct direction for pattern targets at different distances, but made poorer responses when blur was presented in conflict with distance. Binocular viewing improved the accommodative responses in only some infants. PMID- 9205714 TI - Modeling the apparent frequency-specific suppression in simple cell responses. AB - Simple cells in cat striate cortex are selective for spatial frequency. It is widely believed that this selectivity arises simply because of the way in which the neurons sum inputs from the lateral geniculate nucleus. Alternate models, however, advocate the need for frequency-specific inhibitory mechanisms to refine the spatial frequency selectivity. Indeed, simple cell responses are often suppressed by superimposing stimuli with spatial frequencies that flank the neuron's preferred spatial frequency. In this article, we compare two models of simple cell responses head-to-head. One of these models, the flanking-suppression model, includes an inhibitory mechanism that is specific to frequencies that flank the neuron's preferred spatial frequency. The other model, the nonspecific suppression model, includes a suppressive mechanism that is very broadly tuned for spatial frequency. Both models also include a rectification nonlinearity and both may include an additional accelerating (e.g., squaring) output nonlinearity. We demonstrate that both models can be consistent with the apparent flanking suppression. However, based on other experimental results, we argue that the nonspecific-suppression model is more plausible. We conclude that the suppression is probably broadly tuned for spatial frequency and that the apparent flanking suppression is actually due to distortions introduced by an accelerating output nonlinearity. PMID- 9205715 TI - Contrast sensitivity in infants and children with Down syndrome. AB - A new contrast sensitivity (CS) card test was used to estimate contrast sensitivity in 18 infants and children with Down syndrome (DS). The results showed that although the overall shape of the contrast sensitivity functions (CSFs) of the subjects with DS was the typical inverted-U, their CSFs were depressed in comparison to control subjects and this relative loss became larger with increasing spatial frequency. In addition, there was little improvement in CS with age and the mean CSF among children with DS (mean age = 7.3 years) was equivalent statistically to a group of 12-month-olds without DS. The Teller Acuity Cards (TAC) were also used to assess visual acuity in 17 of the 18 children in our sample. The results of these tests showed that their visual acuity (VA) was significantly lower than normal, but was consistent with that extrapolated from each subject's CSF. Taken together with previous anatomical and developmental findings, our results suggest that the deficits in spatial vision among children with DS is due primarily to restricted cortical development, and secondarily, to the additional accommodative and ocular conditions that are prevalent in this population. PMID- 9205716 TI - Chromatic and monochromatic optical resolution in the rainbow trout. AB - The modulation transfer function due to measured longitudinal chromatic aberration was calculated for the otherwise unaberrated eye of the adult rainbow trout (Oncorhynchus mykiss) under daylight conditions assuming light absorption by single retinal cone pigments, and by photopic mechanisms involving interaction between cones. The adult trout eye, with its large immobile pupil, is limited by chromatic aberration to resolution much lower than the diffraction limit, consistent with the low acuity reported for fish. This low resolution can be considered a design trade-off cost of a bright image. The measured monochromatic modulation transfer function is similar to that calculated due to chromatic aberration alone, showing that these independent aberrations are approximately balanced in the fish eye. The effect of changes in receptor length, pigment density, water depth, and pupil size upon the chromatic resolution was calculated. The calculated chromatic modulation transfer function will hold approximately for other teleost eyes with lens larger than about 1 mm. PMID- 9205717 TI - Contrast affects flicker and speed perception differently. AB - We have previously shown that contrast affects speed perception, with lower contrast, drifting gratings perceived as moving slower. In a recent study, we examined the implications of this result on models of speed perception that use the amplitude of the response of linear spatio-temporal filters to determine speed. In this study, we investigate whether the contrast dependence of speed can be understood within the context of models in which speed estimation is made using the temporal frequency of the response of linear spatio-temporal filters. We measured the effect of contrast on flicker perception and found that contrast manipulations produce opposite effects on perceived drift rate and perceived flicker rate, i.e., reducing contrast increases the apparent temporal frequency of counterphase modulated gratings. This finding argues that, if a temporal frequency-based algorithm underlies speed perception, either flicker and speed perception must not be based on the output of the same mechanism or contrast effects on perceived spatial frequency reconcile the disparate effects observed for perceived temporal frequency and speed. PMID- 9205718 TI - The effects of eccentricity and stimulus magnification on simultaneous performance in position and movement acuity tasks. AB - We presented two tasks, spatial interval discrimination and displacement detection, simultaneously in the same location at various eccentricities. The subject was to solve (i) only the spatial interval task; (ii) only the displacement task; or (iii) both tasks simultaneously. With 500 msec stimulus duration, and using the method of spatial scaling, the E2 value (the eccentricity at which stimulus size has to be doubled to maintain performance level) was found to be 0.17-0.39 deg for spatial interval discrimination and 1.0-1.2 deg for displacement detection. These values remained unaffected whether the subject solved one task or two tasks simultaneously. This finding was confirmed using a shorter, 50 msec stimulus duration. As there is no interference between tasks, the mechanisms solving the tasks appear to be functionally independent i.e., operating in parallel at all eccentricities. PMID- 9205719 TI - On the independence of chromatic and achromatic stereopsis mechanisms. AB - The extent to which the processing of stereoscopic depth information can take place separately in colour-contrast-sensitive and luminance-contrast-sensitive mechanisms has been investigated. Contrast thresholds for stereoscopic depth identification (front/back) were measured using 0.5 c/deg Gabor patches. The stimuli possessed different amounts of colour and luminance contrast ranging from isoluminance (red/green) to isochrominance (yellow/black) through intermediate values. Two models for combining chromatic and achromatic stereopsis information were tested. The first (single-pathway) model assumed colour and luminance contrast summation within a single luminance-contrast-sensitive mechanism before stereoscopic judgement. The second (dual-pathway) model assumed probability summation between independent chromatic and achromatic stereopsis mechanisms. The latter model provided the better fit to the data. In providing evidence in favour of an independent chromatic stereopsis mechanism, it was shown that luminance artifacts were unlikely to be the cause of maintained stereopsis at isoluminance. The possible neural substrates of chromatic stereopsis are discussed. PMID- 9205720 TI - Spatio-temporal boundary formation: the role of local motion signals in boundary perception. AB - Spatio-temporal boundary formation (SBF) refers to a perceptual process responsible for perception of moving, bounded surfaces from sequential changes in spatially separated local elements. Previous research has indicated that this process produces perception of global form, continuous boundaries and global motion from spatially and temporally sparse element changes. In the present paper, we sought to distinguish between two classes of models for SBF: form precedes-motion and motion-precedes-form models. Experiment 1 tested the effects of the addition of spurious motion signals, a manipulation that should affect a motion-precedes-form computation but not a form-precedes-motion computation. Shape identification in a 10-alternative forced-choice procedure was disrupted by this manipulation, supporting the former class of models. A particular computational scheme, edge orientation from motion (EOFM) instantiating a motion precedes-form model is described and tested in Experiment 2. The EOFM model should be disrupted when initiating element changes occur in a certain type of sequential order, relative to randomly arranged changes. Sequential changes markedly disrupted performance, supporting this EOFM approach. The results favor motion-precedes-form models of SBF and are consistent with the particular computational scheme proposed. PMID- 9205721 TI - Influence of rod signals on hue perception: evidence from successive scotopic contrast. AB - In successive scotopic color contrast, a colored adapting field induces a hue into a successively presented, purely rod-detected test field. To determine the rod influence on hue perception, a comparison was made, for both spectral matches and hue names, between photopic and scotopic color contrast hues produced by the same adapting fields adjusted to each of the four unique hues. Rod signals evoked hues reflecting each direction of both red/green and blue/yellow hue dimensions. Rod signals differentially strengthened blue relative to red or green hue components under some conditions but not under others. No other differential rod influences on hue were found. PMID- 9205722 TI - A reduced motion aftereffect in strabismic amblyopia. AB - The motion aftereffect was measured using both static and dynamic test stimuli in a group of normal observers and a group of strabismic amblyopes. Amblyopes exhibited a reduced direct aftereffect for both static and dynamic stimuli and only two of the eight amblyopes exhibited any measurable interocular transfer for either test stimulus. It is hard to explain these results in terms of either the known spatial (contrast sensitivity and positional sensitivity) or motion deficits previously reported in amblyopia. These results suggest a primary motion deficit in amblyopia affecting both the static and dynamic motion aftereffects. PMID- 9205723 TI - Object grouping contingent upon background. AB - This study analyzes grouping between singletons (line elements popping out by orientation gradient) when they segregate from textures of uniformly oriented line elements. In the first experiment three adjacent singletons formed a texture bar; in the second experiment the distance between two singletons to be grouped was manipulated. The observer's task was to discriminate the orientation of the global pattern made by the singletons. The results and the explanations suggested are that: (i) an inner gradient within the texture bar (when the singletons are reciprocally orthogonal) operates only at short distances and enhances discrimination, indicating an initial stage of texture segregation based on local processing. (ii) Spatial interactions between parallel singletons are present at short distances and reduce discrimination. (iii) An interruption of background flow (directed along the orientation of background line elements) produced by the grouped pattern when orthogonal to it, enhances discrimination; this effect is present at both short and large distances between singletons, indicating a global process. (iv) Spatial interactions are present between parallel singletons even at large distances and independently of background orientation, suggesting that grouping generates a figural context within which features to be bound together interact. Moreover, flow interruption and figural context were absent in a detection task, thereby suggesting their specific involvement in grouping and figure binding. Overall, the results may indicate that grouping operates on already segmented line elements, across different orientations and over both short and long distances in between. PMID- 9205724 TI - Vernier acuity with plaid masks: the role of oriented filters in vernier acuity. AB - Superimposition of oriented grating masks on vernier targets results in bimodal patterns of vernier threshold elevation, with peaks occurring on either side of vernier target orientation. These bimodal masking effects suggest a contribution to vernier acuity from spatial filters tuned to orientations on either side of the target. We report similar bimodal threshold elevation with plaid masks composed of symmetrically oriented pairs of gratings. Since filters oriented to either side of the vernier stimulus will be affected similarly by plaid masks, it is unlikely that threshold elevation reflects disruption of relative filter activity that is used to code for change in target orientation. Instead, the results support the proposition that misalignments are detected on the basis of differential (i.e. absolute rather than relative) activity of spatial filters. Our plaid-mask data also rule out the possibility that: (i) "off-channel" looking; or (ii) detection of orientation shifts (e.g. tilt illusions), underlie bimodal masking effects. The finding that weak bimodal threshold elevation occurs with dot targets separated by 40 min arc further suggests that the mechanisms involved in detecting misalignments over large regions [possibly collator/collector-type mechanisms] also do so via analysis of their differential activity. PMID- 9205725 TI - Four issues concerning colour constancy and relational colour constancy. AB - Four issues concerning colour constancy and relational colour constancy are briefly considered: (1) the equivalence of colour constancy and relational colour constancy; (2) the dependence of relational colour constancy on ratios of cone excitations due to light from different reflecting surfaces, and the association of such ratios with von Kries' coefficient rule; (3) the contribution of chromatic edges to colour constancy and relational colour constancy; and (4) the effects of instruction and observer training. It is suggested that cognitive factors affect colour constancy more than relational colour constancy, which may be an inherently more robust phenomenon. PMID- 9205726 TI - Predictive smooth pursuit of complex two-dimensional trajectories demonstrated by perturbation responses in monkeys. AB - Two-dimensional sum-of-sines waveforms were pursued by the eye with very small phase delays compared with visual feedback delays estimated in the same monkeys. Processing delays in making smooth corrections averaged 90 msec after infrequent right-angle perturbations from a circular trajectory. These feedback delays were much larger than component phase delays during pursuit that averaged: 10 msec for sinusoids, 3 msec for circles, 20 msec for sum-of-two-sines trajectories, and 19 msec for sum-of-three-sines trajectories. This suggests that predictive control can play a strong role during tracking for a variety of simple and complex target trajectories. PMID- 9205727 TI - Listing's plane dependence on alternating fixation in a strabismus patient. AB - Listing's law of the eye is one of the best studied findings in motor control, but its functional meaning is still incompletely understood and its status in neurological disorders and in strabismus is almost entirely unknown. We investigated the mechanisms underlying Listing's law and its possible clinical relevance. The dual magnetic search coil technique was used to record three dimensional binocular eye movements in a stereoblind strabismic patient with good visual acuity in both eyes and capable of voluntarily alternating fixation. This technique yielded an accurate, objective and simultaneous measure of ocular misalignment in three dimensions and showed that the squint angle depended on which eye was fixating. Saccadic eye movement data throughout the oculomotor range were used to fit Listing's plane. Listing's primary position and the thickness of the plane for each eye were calculated for three different fixation conditions. For comparison, control measurements were taken from four normals. In the patient, no large deviations from normal values for the thickness of Listing's plane and the confidence limits of the Listing primary position were found. The most remarkable abnormality was that the orientation of Listing's plane depended on which eye was fixating. Both the change in ocular misalignment and the shift of Listing's primary positions observed when changing fixation are probably linked to accommodation-related vergence. Despite repeated surgery at early age, the patient had well-defined Listing planes for both eyes, but their alignment during left-eye fixation was abnormal. The obedience to Listing's law may reflect a strategy which minimizes muscular effort in each eye separately. The abnormal fixation-condition dependence is probably due to an aberrant coupling with vergence. PMID- 9205728 TI - Specificity of saccadic adaptation in three-dimensional space. AB - The saccadic system is known to exhibit a considerable degree of short-term plasticity. Earlier studies have shown that saccadic adaptation, rather than being a global process affecting all saccades equally, has a certain degree of spatial resolution. Its localized nature has become apparent from studies in the frontal plane which have shown that short-term saccadic adaptation, induced along a given meridian, transfers to only a limited range of neighbouring directions. Considering that most natural gaze shifts also have a depth component, we investigated whether the directional specificity of the saccadic adaptive system can be generalized to three-dimensional (3-D) space. Binocular eye movements were recorded in seven subjects while they made saccades to visual stimuli in the horizontal plane of regard. Experiments began by recording baseline saccades, all starting from the same fixation point to either a farther target (far saccades) or an equally eccentric nearer target (near saccades). Next, by displacing the target intra-saccadically in opposite directions in alternating far and near trials, we attempted to simultaneously reduce the gain of the far saccades while increasing the gain of the near saccades. These experiments, aimed at eliciting a state of differential gain, were specifically designed to adapt only the saccadic response, since targets were shifted along corresponding iso-vergence circles. To investigate the effect of varying the radial direction difference, similar differential gain adaptation experiments were conducted in the frontal plane for saccades along two different meridians. Our results show that when the saccadic system is pressured, it is capable of adopting different gains simultaneously for equal-direction saccades to different depth planes. Similarly, opposite gain adaptation can also be achieved in the frontal plane, but only if radial saccade directions are sufficiently separated. The fact that short-term saccadic adaptation can be shown to be directionally specific in two perpendicular planes suggests that the adaptation process is restricted to a limited volume of 3-D oculomotor space. PMID- 9205729 TI - Neural network model of short-term horizontal disparity vergence dynamics. AB - We present a neural network model of short-term dynamics of the human horizontal vergence system (HVS) and compare its predictions qualitatively and quantitatively with a large variety of horizontal disparity vergence data. The model consists of seven functional stages, namely: (1) computation of instantaneous disparity; (2) generation of a disparity map; (3) conversion of the disparity into a velocity signal; (4) push-pull integration of velocity to generate a position signal; (5) conversion of the position signal to motoneuron/plant activity for each eye; (6) gating of velocity overdrive signal to motoneuron/plant system; and finally (7) discharge path for position cells. Closed-loop (normal binocular viewing) symmetric step and staircase disparity vergence data were collected from three subjects and model parameters were determined to quantitatively match each subject's data. The simulated closed-loop as well as open-loop (disparity clamped viewing) symmetric step, sinusoidal, pulse, staircase, square and ramp wave responses closely resemble experimental results either recorded in our laboratory or reported in the literature. Where possible, the firing pattern of the neurons in the model have been compared to actual cellular recordings reported in the literature. The model provides insights into neural correlates underlying the dynamics of vergence eye movements. It also makes novel predictions about the human vergence system. PMID- 9205730 TI - Early impairment of foveal magno- and parvocellular pathways in juxta chiasmal tumours. AB - Foveal pathway visual function was assessed in 11 patients having tumours extending into the suprasellar region but without evidence of visual impairment as assessed by visual acuity and Bjerrum screen campimetry. Psychophysical and routine visual evoked potential (VEP) measurements were obtained from the eye ipsilateral to the maximal suprasellar extension. The sensitivity of luminance and chromatic pathways was assessed psychophysically by measuring increment thresholds for white and red flashes of light presented on a white adapting field. Temporal sensitivity was assessed psychophysically by measuring threshold modulation sensitivity for sinusoidally modulating stimuli (de Lange attenuation characteristic). The patient group showed approximately equal significant psychophysical losses in chromatic, luminance and temporal sensitivities relative to normal controls. Midline VEP P100 latencies of the patient group did not significantly differ from those of the normal control group. It is concluded that tumours extending into the suprasellar region can cause foveal pathway dysfunction affecting both magno- and parvocellular pathways, even in the presence of normal visual acuity and fields suggesting a more widespread and insidious abnormality of the visual pathways in this condition than previously thought. PMID- 9205731 TI - Perioperative mortality in Germany. PMID- 9205732 TI - Oxygen consumption by splanchnic tissues in wethers consuming ad libitum different proportions of bermudagrass and ryegrass-wheat. AB - Crossbred wethers (n = 18, 7.5 month of age and 31 +/- 0.8 kg) were used in a 23 day experiment to determine effects of ad libitum consumption of diets differing in proportions of coarsely chopped bermudagrass and ryegrass-wheat hay (0, 33, 67 and 100%) on oxygen consumption by splanchnic tissues. Bermudagrass and ryegrass wheat were 9 and 13% CP and 78 and 71% NDF, respectively. Intake of dry matter (1.03, 0.92, 0.92 and 0.76 kg/d) and digestible energy (13.5, 10.7, 10.6 and 8.2 MJ/d for 0, 33, 67 and 100% bermudagrass, respectively) changed linearly and cubically (P < 0.05) as bermudagrass level increased. Consumption of oxygen by the portal-drained viscera tended to decrease linearly (P = 0.14) with increasing bermudagrass (182, 154, 156 and 137 mM/h), and hepatic oxygen consumption decreased linearly (P < 0.05) and changed cubically (P = 0.07; 150, 113, 116 and 103 mM/h for 0, 33, 67 and 100% bermudagrass, respectively). Splanchnic tissue energy consumption expressed as a percentage of digestible energy intake increased linearly (P = 0.08) with increasing bermudagrass (24.0, 27.6, 28.6 and 33.2% for 0, 33, 67 and 100% bermudagrass, respectively). In conclusion, the level rather than presence alone of different grass sources consumed ad libitum affected energy use by the splanchnic bed, and as a percentage of digestible energy intake splanchnic bed energy consumption increased with increasing dietary bermudagrass level and decreasing digestible energy intake. PMID- 9205733 TI - Absorption and metabolism of nivalenol in pigs. AB - The absorption and metabolism of nivalenol (NIV) were studied in pigs fed 0.05 mg NIV/kg BW, twice daily. Blood samples were taken during the first and third day, through catheters in the hepatic portal vein and peripheral mesenteric artery. Nivalenol was detected in most of the earliest blood samples, taken twenty minutes after the start of feeding. During 7.5 hrs after feeding, 11-43% of the NIV dose was absorbed. The systemic peak concentrations were 3-6 ng NIV/ml, mostly occurring 2.5-4.5 h after feeding. Sixteen hours after feeding, NIV was still being absorbed from the intestine, and the systemic concentrations were 1-3 ng NIV/ml. Nivalenol was mainly excreted in faeces, which contained concentrations up to 3.2 mg NIV/kg. No metabolites of NIV were found in plasma, urine, and faeces, either as glucuronic acid or sulphate conjugates, or as de epoxy-NIV, indicating a lack of metabolism. The feeding of NIV did not cause feed refusal, and measured clinical plasma parameters were within the normal ranges. PMID- 9205734 TI - Effect of maduramicin and monensin on survival of Lactobacillus salivarius 51R administered in the crop and caeca of young chickens. AB - A rifampicin-resistant Lactobacillus salivarius 51R was administered orally to newly hatched broiler chickens. The resistance to rifampicin enabled us to differentiate the organism administered from indigenous strains. One day after inoculation, Lactobacillus salivarius 51R dominated among lactobacilli in the crop and caeca of all inoculated chickens, even in those ones receiving maduramicin and monensin at 5 and 100 mg per kg of feed mixture, respectively. Coliform counts in both crop and caeca of inoculated chickens were significantly lowered on the first day after treatment. Also, counts of the crop enterococci were decreased in inoculated chickens. Rifampicin-resistant lactobacilli were still present in high numbers in the crop and caecal contents of inoculated chickens sampled 5 days after inoculation. Differences in counts of total lactobacilli, coliform bacteria, and enterococci were mostly nonsignificant in these samples. Our results demonstrate that (i) bacterial counts in the chicken gut were influenced by probiotic Lactobacillus administration, and (ii) chicken lactobacilli are resistant to ionophore coccidiostats under in vivo conditions. PMID- 9205735 TI - Omega-3 fatty acids in pig nutrition: implications for zootechnical performances, carcass and fat quality. AB - The extent of incorporation of dietary alpha-linolenic acid -readily available in linseed- in pig diets, in view of repercussions on zootechnical performance, carcass and fat quality of pigs, is investigated. Ninety hybrid pigs (Pietrain x Seghers hybrid cross, 41 barrows and 49 gilts), divided in three comparable groups, were fed ad libitum three experimental diets, containing respectively 4 g, 7 g and 10 g alpha-linolenic acid per kg feed, originating from linseed. The increase of polyunsaturated fatty acid content in the feed (11.9 g, 15.2 g and 18.8 g per kg feed, resp.) could almost be completely attributed to the variation in alpha-linolenic acid. Despite several anti-nutritional factors, present in linseed, zootechnical performance was not affected by the diet. However, carcass quality, in terms of lean meat % and conformation, was less favourable for the highest linseed level compared to the intermediate level. Loin fat thickness was not influenced by the fat source in the diet. More pronounced was the effect of sex on zootechnical and carcass parameters: gilts showed a lower feed intake and weight gain, resulting in a more favourable feed conversion ratio, a thinner backfat layer, a higher meat content and a superior conformation, compared to the barrows. The linolenic acid content in the backfat increased from 3.1 g to 6.8 g per 100 g of total fatty acids for the barrows and from 3.4 g to 7.0 g per 100 g of total fatty acids for the gilts. A significant positive correlation was found between the live weight at slaughter and C16:0 and C18:0 content in the backfat; C18:2 and C20:4 content, on the contrary, were negatively correlated with the live weight at slaughter. A more unsaturated fatty acid pattern of the backfat, as a result of higher C18:3 levels in the feed, resulted in higher TBA-values (thiobarbituric acid), without occurrence, though, of off-odours during the fat thawing. Thus PUFA content in the backfat reached a maximum of respectively 18 g and 19 g per 100 g of total fatty acids for the barrows and the gilts, without implications for the consistency of the fat. PMID- 9205736 TI - Preileal digestibility of coconut fat and soybean oil in horses and their influence on metabolites of microbial origin of the proximal digestive tract. AB - Three horses (approximately 190 kg BW) fitted with a permanent fistula at the end of the jejunum were used. To a control diet (1/3 hay, 2/3 mixed feed) one of two fat types (coconut fat or soybean oil) were added at 2 levels resulting in fat intakes of 0.1 g (control diet) to 0.5 or 1 g/kg BW 0.5 d, respectively. Each experimental period consisted of 2 weeks adaptation, 2 days of breath tests (before and hourly after the morning meal) and 5 days sampling of chyme. Crude fat, crude protein, concentrations of organic acids (SCFA, lactic acid), pH, and the minerals calcium, magnesium and phosphorus were determined in the chyme; H2 and CH4 in the expired air. The following results were obtained: 1) Fat feeding significantly (P < 0.01) stimulated (independent of amount or kind of fat) the jejunoileal flow of chyme. 2) Preileal fat digestibility increased significantly (P < 0.01) from 30-38% during the control periods to 73-80% (moderate fat intake) and 82-86% (high fat intake). Differences between the fat sources were not significant. 3) Fat addition resulted dose dependent in a reduction (P < 0.05) of lactic acid as well as SCFA concentrations of chyme (at 5th h postprandial). 4) Fat intake caused a reduction in the H2-concentration of the exhaled air (P < 0.05). Such effect was not found with the CH4-concentration, except the high soybean oil level which tended to reduce the concentration. 5) The addition of fat had no significant effects on preileal net absorption of magnesium and calcium, whilst the net secretion of phosphorus significantly increased (P < 0.01). 6) The preileal protein digestibility (control periods 48-53%) was slightly decreased (P < 0.05), due to the fat inclusion. PMID- 9205737 TI - Ebrotidine. A new generation H2-receptor antagonist and gastroprotective agent. Introduction. PMID- 9205738 TI - Synthesis and assessment of formamidines as new histamine H2-receptor antagonists. AB - Four series of compounds whose substructure contains a formamidine functionalized as a novel group in the chemistry of histamine H2-receptors have been synthesized. Series design, synthesis and pharmacological data including inhibition of histamine-stimulated acid secretion, inhibition of acid secretion p.o. and pA2 are reported. N-[(E)-[[2-[[[2](Diaminomethylene)amino]-4-thiazolyl] methyl]thio]ethyl]amino]methylene]-4-bromo-benzenesulfonamide (ebrotidine, CAS 100981-43-9, FI-3542) was selected for further research. PMID- 9205739 TI - Physicochemical properties, analytical determinations and stability of ebrotidine. AB - Ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]-4-thiazolyl]methyl] thio]ethyl]amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) is a new H2-receptor antagonist with a potent antisecretory activity and evidenced gastroprotection. This paper describes its physicochemical properties, spectroscopy for its structural identification, detection methods for its organic and inorganic impurities, purity quantitation and stressed degradation and stability tests in solid and solution forms in order to know the behaviour of the test substance against certain experimental conditions. The results obtained indicate that ebrotidine is stable for over 3 years normal storage conditions (25 degrees C/75% RH). As ebrotidine was not found to be hygroscopic or particularly photosensitive, no special storage precautions are required. PMID- 9205740 TI - Histamine H2-receptor antagonist action of ebrotidine. Effects on gastric acid secretion, gastrin levels and NSAID-induced gastrotoxicity in the rat. AB - The antagonism of histamine H2-receptors by ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl]methyl]thio]ethyl ] amino]methylene]-4 bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) was assessed on isolated guinea-pig right atrium. The dose-response curves obtained by histamine on the positive chronotropic effect in guinea-pig atrium were displaced to the right in parallel depending on the concentration of ebrotidine and ranitidine without change in the maximum response with pA2 values of 7.12 and 7.26, respectively. The slope of the regression line of log (DR-1) against log ebrotidine concentration was not significantly different from unity: 0.96 (95% confidence limits: 0.89-1.03). These results indicate that ebrotidine is a competitive H2 receptor antagonist. Following intravenous administration to rats, ebrotidine inhibited histamine- and pentagastrin-stimulated acid secretion in a dose dependent manner, ED50 being 0.21 and 0.44 mg/kg, respectively. After oral administration to fasting rats 3 h before their sacrifice, ebrotidine decreased the total acid contents of the stomach in a dose-dependent manner, ED50 being 7.5 mg/kg. After a single dose of 100 mg/kg in fasting rats, ebrotidine increased significantly serum gastrin levels within 2 and 5 h after administration, but 8 h after administration serum gastrin levels returned to normal values. In contrast, ranitidine at a single oral dose of 100 mg/kg increased serum gastrin levels more markedly within 2 and 5 h after administration, while after 8 h, this increase still persisted although without significant differences with respect to control, and after 24 h levels returned to normal values. Both ebrotidine and ranitidine were administered orally at a dose of 100 mg/kg for 26 days showing significant increments in plasma gastrin levels 5 h after administration. Such increments were not so marked after ebrotidine and normal values were attained at 24 h after administration. The results obtained after repeated oral administration for 15 days of ebrotidine and ranitidine at the doses of 15 and 50 mg/kg demonstrated that ebrotidine did not increase significantly serum gastrin levels with respect to control 2 h after administration, and no dose-related effect was observed. In contrast, ranitidine increased serum gastrin levels significantly and in a dose dependent manner with respect to control group. ED50 values of ebrotidine obtained in the experiments on the prevention of NSAID-induced gastrotoxicity in the rat were 12.2, 12.5, 11.5 and 9.8 mg/kg against diclofenac, ketoprofen, indometacin and naproxen, respectively. ED50 values of ranitidine were of the same order: 20.6, 13.9, > 50 and 15.1 mg/kg. PMID- 9205741 TI - Action of ebrotidine, ranitidine and cimetidine on the specific binding to histamine H1- and H2-receptors. AB - Ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)-4-thiazolyl]methyl]thio] ethyl]amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542), a selective H2-receptor antagonist, has proved to competitively inhibit the positive chronotropism induced by histamine in isolated guinea pig atrium. The affinity of ebrotidine to histamine H1- and H2-receptors through the displacement of 3H-pyrilamine and 3H-thiotidine binding to guinea pig cerebellum and brain cortex membranes was investigated. Ebrotidine displaced 3H-thiotidine specific binding to histamine H2-receptors (Ki: 127.5 nmol/l), showing a higher affinity (p < 0.05) than ranitidine (Ki: 190.0 nmol/l) and cimetidine (Ki: 246.1 nmol/l). None of the three substances displaced 3H-pyrilamine binding to H1-receptors (Ki: > 5000 nmol/l). The results showed that ebrotidine is a drug with a high affinity for H2 receptors, higher than cimetidine and ranitidine. PMID- 9205743 TI - Study on the increment of the amount of gastric mucus in rats after repeated-dose administration of ebrotidine. AB - Ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]-4 thiazolyl]methyl]thio]ethyl ] amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) is a novel H2-receptor antagonist that also exhibits a potent gastroprotective action against ethanol damage. This study was designed to ascertain under physiological conditions the effect of ebrotidine on the secretion of gastric mucus, probably the main component of the mucosal barrier. Two groups of 20 rats each were given a daily oral dose of 10 or 35 mg/kg ebrotidine, respectively, for 17 days. A third group of 20 rats was used as a control. Once the administration period had concluded, the animals were killed and their stomachs were removed and processed by the periodic acid-Schiff (PAS) histochemical method, selective for mucopolysaccharides. PAS-positive areas exhibited a characteristic carmine colour, allowing morphometric study by computerized image analysis. All the histological sections studied were from the same region of the stomach. A significant increase in the PAS-positive area corresponding to glandular mucus was found in all treated groups. This action is consistent with an increased secretion of mucopolysaccharides and represents one of the main mechanisms of the cytoprotective action of ebrotidine. PMID- 9205742 TI - Effect of ebrotidine on ethanol-induced gastric mucosal damage in the rat. Comparative study with other H2-receptor antagonists. AB - Ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]-4 thiazolyl]methyl]thio]ethyl ] amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) is a novel H2-receptor antagonist with additional gastroprotective effect. The gastroprotective effect of oral doses of ebrotidine against ethanol-induced mucosal damage in female rats was compared with the effect of cimetidine, ranitidine and famotidine. Macroscopically, ebrotidine showed dose-dependent inhibition of the lesion, with significant differences from that of controls at doses of > or = 12.50 mg/kg (ED50 was 26.54 mg/kg). None of the other drugs tested showed gastroprotective effect under the same conditions. The histopathological study revealed significant reduction in the number of deep and superficial ulcers in ebrotidine-treated animals. The gastroprotective effect of ebrotidine is patent even in the presence of indometacin suggesting that prostaglandins play a rather negligible role in gastroprotective action. These results suggest that ebrotidine may be more useful than the classically known H2 antagonists in the treatment of peptic ulcers. PMID- 9205744 TI - Gastroprotective properties of ebrotidine. A review. AB - Ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]-4-thiazolyl] methyl]thio]ethyl]amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) is a new antiulcer drug which combines the properties of an H2-receptor antagonist with those of a cytoprotective agent. The cytoprotective properties of ebrotidine are not dependent upon endogenous prostaglandin generation, but stem from the ability of the drug to induce mucosal responses manifested in the enhanced physicochemical characteristics of mucus gel. These include the increase in mucus gel dimension, viscosity, hydrophobicity and hydrogen ion retardation capacity. Improvements in mucus gel protective qualities with ebrotidine are directly related to the ability of the drug to enhance the synthesis and secretion of sulfo- and sialomucins and phospholipids of gastric mucus and to promote mucin macromolecular assembly. An equally important property of ebrotidine in promotion of ulcer healing is its capability to enhance the gastric mucosal expression of integrin receptors for the interaction with proteins of the extracellular matrix such as laminin. Furthermore, the accelerated ulcer healing with ebrotidine is reflected in a marked increase in the mucosal expression of EGF and PDGF receptors. The drug has also been shown to modulate the processes associated with cell cycle progression during ulcer healing, and is known to protect the gastric epithelial integrity from calcium imbalance. Thus, ebrotidine, unlike other H2-blockers, has a unique ability to promote the event essential for mucosal repair and the maintenance of mucosal integrity. These features make ebrotidine a drug of great potential in the treatment of ulcer disease. PMID- 9205745 TI - Involvement of endogenous nitric oxide and sulfhydryl compounds in ebrotidine induced gastroprotection. AB - The role of endogenous nitric oxide and sulfhydryl compounds in the prevention by ebrotidine (N-[(E)- [[2-[[[2-[(diaminomethylene)amino]-4 thiazolyl]methyl]thio]ethyl]amino] methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) (100 mg/kg i.g.) of ethanol-induced gastric damage in rats was demonstrated. When the animals were pretreated with N-nitro-L-arginine methyl ester, an inhibitor of nitric oxide synthase, at the dose of 10 mg/kg i.v., the mucosal lesions were aggravated and the gastroprotective action of ebrotidine decreased from 85% to 24%. This decrease in ebrotidine protection was antagonized by L-arginine (200 mg/kg i.v.), the lesion inhibition rate being 69%. D-arginine (200 mg/kg i.v.) was ineffective and the inhibition afforded by ebrotidine was only 14%. Pretreatment with N-ethylmaleimide, a sulfhydryl blocker, at the dose of 50 mg/kg s.c., increased the mucosal lesion induced by ethanol, and the gastroprotective action of ebrotidine decreased from 75% to 9%. These results suggest that endogenous nitric oxide and sulfhydryl compounds play a crucial role in the gastroprotective activity of ebrotidine. PMID- 9205746 TI - In vitro anti-Helicobacter pylori activity of ebrotidine. AB - The in vitro anti-Helicobacter pylori (H. pylori) activity of ebrotidine (N-[(E) [[2-[[[2-[(diaminomethylene) amino]-4 thiazolyl]methyl]thio]ethyl]amino]methylene]-4-bromo- benzenesulfonamide, CAS 100981-43-9, FI-3542) versus ranitidine, and their effect on the susceptibility to the antimicrobial agents used in H. pylori eradication were investigated. Assessment was performed by determining the minimum inhibitory concentrations (MIC) against 9 strains of H. pylori, 8 from clinical source and 1 from the American Type Culture Collection (ATCC 43504), in Mueller-Hinton solid media plus 7% blood. The concentration of inocula was 10(7) CFU/ml, incubation was performed at 37 degrees C in microaerophilic atmosphere, and results were read after 5 days of growth. Ebrotidine gave a mean MIC value of 75 micrograms/ml, while that for ranitidine was > 1000 micrograms/ml. Ebrotidine at 100 micrograms/ml enhanced the activity of the antimicrobials studied as follows: erythromycin 3 times, tetracycline 1.1 times, amoxicillin 3 times, metronidazole-sensitive strains 9 times and clarithromycin 5 times. Ranitidine had no effect on the MIC of the antibiotics even at 500 micrograms/ml. PMID- 9205748 TI - Pharmacokinetics of ebrotidine in rats and dogs. AB - The pharmacokinetics of ebrotidine (N-[(E)-[[[2-[(diaminomethylene)amino] -4 thiazolyl]methyl]thio]ethyl] amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) was studied in the rat and dog. After oral (agar suspension) and intravenous administration at 10 mg/kg to rats, ebrotidine was rapidly absorbed. Cmax values averaged 0.498 microgram/ml attained at tmax = 30 min. Distribution was fitted to a two-compartmental model with t1/2 beta = 1 h (i.v.). Clearance (Cl) was 29 ml/min.kg and volume of distribution (Vdss) was 1852 ml/kg. Absolute bioavailability was 22% of the dose administered. After oral (the same tablet formulation as that used for clinical trials) and intravenous administration at 150 mg and 25 mg, respectively, to dogs, absorption of ebrotidine was relatively rapid. Cmax values averaged 2,170 micrograms/ml attained at tmax = 2 h. Distribution was fitted to a two-compartmental model with t1/2 beta = 2.8 h (i.v.). Clearance (Cl) was 600 ml/h.kg and volume of distribution (Vdss) was 1000 ml/kg. Absolute bioavailability, which is variable in this type of drugs, ranges from 29% to 64% of the dose administered. PMID- 9205747 TI - Anti-Helicobacter pylori activities of ebrotidine. A review of biochemical and animal experimental studies and data. AB - Infection with Helicobacter pylori (H. pylori) is now recognized as a major factor in the pathogenesis of gastric disease, and the successful therapy regimens require a combination of H2 blockers with gastroprotective and antimicrobial agents. Ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene) amino]-4 thiazolyl] methyl]thio]ethyl]amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) is the only drug combining acid-suppressant activity with remarkable gastroprotective and anti-H. pylori properties. The drug not only displays a potent anti-H. pylori activity alone, but also exerts a strong potentiating effect on the efficacy of antimicrobial agents commonly used for H. pylori eradication, and the successful ulcer therapy with ebrotidine induces a significant (4-fold) increase in the H. pylori aggregation titer of gastric mucin. Moreover, the drug exhibits a strong inhibitory effect on H. pylori urease activity, the extent of which exceeds that of ranitidine, omeprazole and lansoprazole. Ebrotidine has also been demonstrated to exert a potent inhibitory action on the enzymatic activities directed towards mucus perimeter of gastric mucosal defense, causing a marked inhibition of H. pylori protease, lipase and phospholipase A2 activities. Another important property of ebrotidine is its ability to efficiently counteract the disruptive effects of H. pylori lipopolysaccharide on the integrity of gastric epithelium. This includes countering the interference by the lipopolysaccharide in mucosal integrin receptor interaction with proteins of extracellular matrix and the reversal of H. pylori disruptive effect on the binding of mucin to its gastric epithelial receptor. Furthermore, most recent data indicate that ebrotidine has the ability to reverse the impairment caused by H. pylori in feedback inhibition of gastrin release by somatostatin. This activity of ebrotidine apparently stems from the drug's ability to counter the untoward effect of H. pylori on the binding of somatostatin to its specific receptor on the gastric mucosal G-cells. The unique combination of acid suppressant, gastroprotective and anti-H. pylori activities makes ebrotidine a drug of choice in the treatment of gastric disease caused by H. pylori. PMID- 9205749 TI - Metabolism of ebrotidine. A review. AB - A hypothetical metabolic pathway for ebrotidine (N-[(E)-[[(2-[[[2 [(diaminomethylene]amino]-4-thiazolyl] methyl]thio]ethyl]amino]methylene-4-bromo benzenesulfonamide, CAS 100981-43-9, FI-3542) has been proposed on the basis of previous data on the metabolism of other H2-receptor antagonists as well as on in vitro degradation assays of ebrotidine. Its potential metabolites have been synthesized and characterized, and their presence in human urine has been investigated by high-performance liquid chromatography (HPLC). Analytical-scale HPLC allowed the identification of metabolites by means of their retention time and UV spectrum, while semipreparative-scale HPLC allowed their identification through FT-IR and 1H-NMR. Mass spectrometry using atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI) for HPLC-MS coupling allowed the identification of all metabolites in human urine. The quantitative determination of ebrotidine and its derivatives has been performed according to a newly designed method which consisted of a liquid-liquid extraction in a basic medium followed by reversed-phase HPLC with ion-pair formation. This method was sensitive, precise and no chromatographic interferences with other drugs which might be administered in combination with ebrotidine were observed. In order to elucidate the excretion of ebrotidine, the analytical method was applied to the analysis of the urine collected from 2 healthy volunteers 96 h after receiving 400 mg of ebrotidine. PMID- 9205750 TI - Acute toxicity studies of ebrotidine. AB - The acute toxicity of two formulations of ebrotidine (N-[N-[(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl] methyl]thio]ethyl]amino]methylene]-4-bromo benzenesulfonamide, CAS 100981-43-9, FI-3542) was studied in rats and mice by different routes of dosing: oral and intraperitoneal routes in rats and mice (suspension), intravenous and oral routes in mice and intravenous route in rats (injectable solution). LD50 values for the oral route were indeterminable in all cases. For the intraperitoneal route. LD50 values were 316 mg/kg (rat) and 366 mg/kg (mouse), and for the intravenous route LD50 values were 100 mg/kg (rat) and 107 mg/kg (mouse). PMID- 9205751 TI - Subacute toxicity of ebrotidine in rats and dogs. AB - Subacute toxicity studies of ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl]methyl] thio]ethyl]amino]methylene]-4-bromo benzenesulfonamide, CAS 100981-43-9, FI-3542) were performed in Spragu-Dawley rats and Beagle dogs. Both animal species were administered with the same dose levels (50, 200 and 500 mg/kg) for 4 and 7 weeks, respectively. In a previous 4 week subacute toxicity study in the rat, ranitidine and cimetidine at 500 mg/kg were used as reference drugs. The results indicated that ebrotidine was well tolerated at 50 mg/kg, while there were dose-related effects at 200 and 500 mg/kg. Probably due to its pharmacokinetics, ebrotidine was more toxic in dogs than in rats, since the most severe effects were the death or sacrifice in extremis of two dogs from the high dose group which had undergone rectal prolapse, while no deaths occurred in the rats. The changes that were very likely related to treatment (500 mg/kg) were a lower weight in both species, a slight decrease of hematocrit and red blood cells in rats, single increments of transaminases, alkaline phosphatase and lactate dehydrogenase in dogs (some animals of the 200 mg/kg dose group were also affected) and a higher liver weight. These effects with a few exceptions were found to be common to cimetidine and ranitidine. PMID- 9205752 TI - Chronic toxicity of ebrotidine in rats and dogs. AB - The results obtained in the chronic toxicity studies of ebrotidine (N-[(E)-[[2 [[[2-[(diaminomethylene)amino]-4-thiazolyl] methyl]thio]ethyl]amino]methylene]-4 bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) by oral route in rats and in Beagle dogs are reported. Rats were administered for 6 months and dogs for 12 months. The doses were 50, 200 and 500 mg/kg in rats and 50, 200 and 400 mg/kg in dogs. The dose of 400 mg/kg was reduced to 350 mg/kg after 3 months of treatment, due to its toxicity. The effects probably related to the administration of ebrotidine were as follows: three dogs from the high dose group died after 3, 4.5 and 8 months of treatment (in rat there was no mortality); occult blood in faeces; lower weight gain in the high dose group (in rats only females were affected); lower food consumption in rats from the high dose group (and also females from the middle dose group); reduction of erythrocyte count and packed cell volume, only in rats and at the end of the study; alkaline phosphatases increment in rats and dogs; proteinemia decrease in rats; and a tendency to decrease in the testicular weight, which was not statistically significant (p > 0.05). The only histopathological changes observed were moderate erosions or ulcerations in the intestinal mucosa of some dogs from the high dose group. These effects coincide with those published for other competitive H2-receptor inhibitors. The maximum toxic effect-free level was 50 mg/kg for both rats and dogs, which provides a wide safety margin with respect to the therapeutic dose. PMID- 9205753 TI - Toxicity of ebrotidine on reproduction. Toxicity on fertility and general reproductive performance, embryo-fetal toxicity and peri- and postnatal toxicity. AB - Reproduction toxicity studies of ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl]methyl] thio]ethyl]amino]methylene]-4-bromo benzenesulfonamide, CAS 100981-43-9, FI-3542) are presented in this paper. Rats dosed with 50, 200 and 500 mg/kg p.o. of ebrotidine were used for the fertility and peri- and postnatal toxicity studies, and rabbits dosed with 25, 100 and 250 mg/kg and rats dosed with 50, 200 and 500 mg/kg of ebrotidine were used for the embryotoxicity study. The fertility study was designed in accordance with a 2 generation study protocol. The results showed that ebrotidine did not interfere with male and female gametogenesis, fertility, organogenesis, postnatal development and lactation in F0 or F1 animals. Only general or non-specific effects were attributed to treatment, such as a lower weight gain in parents or fetuses in rats, or a somewhat slower bone calcification in rats, which was shown to be recoverable and had no peri- or postnatal repercussions. Neither did the fertility study reveal a possible longer duration of gestation nor did the peri- and postnatal study show a lower weight of the F1 offspring. There was only an increase in rabbit embryonic mortality, probably related to some cases of abortion at the high dose. No potential antiandrogenic effect on the reproductive function has been found. Among the different doses used in both animal species, the maximum toxic effect-free dose was that of 25 mg/kg. PMID- 9205754 TI - Genotoxicity studies on ebrotidine. AB - Five genotoxicity studies on ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl]methyl]thio] ethyl]amino]methylene]-4-bromo benzenesulfonamide, CAS 100981-43-9, FI-3542), including at least four of the battery of tests recommended by toxicological regulatory guidelines for new drugs, were conducted. These tests were the Ames test for determination of bacterial gene mutations, sex-linked recessive lethal mutation test in Drosophila for gene mutations in eukaryotic systems, in vitro chromosome aberration test and micronucleus test for evaluation of structural and numerical aberrations, and sister chromatid exchange frequency test for assessment of non-specific damage to chromatin. Negative and positive controls were used in all the experiments. The effects were investigated in the absence or presence of metabolic activation by S 9 microsomal fraction from rat liver homogenate. A dose range toxicity study was also performed to determine the dosage levels or concentrations to be tested for the assessment of genotoxic effects. None of the tests showed a significant increase in the genotoxic parameters, both in vitro and in vivo in somatic or germ cells. It is, therefore, concluded that ebrotidine has not caused mutagenic or clastogenic effects in any of the experimental systems tested. PMID- 9205755 TI - Carcinogenicity studies on ebrotidine. AB - The results from two carcinogenicity studies on ebrotidine (N-[2-(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl] methyl]thio]ethyl]amino]methylene]-4-bromo benzenesulfonamide, CAS 100981-43-9, FI-3542) conducted in mice and rats are reported. Oral doses of 50, 200 and 500 mg/kg were administered to mice for 18 months and 50, 200 (150), 300 and 500 mg/kg were administered to rats for 24 months. The study design was prepared according to EEC guidelines, and the recommendations by the International Agency for Research on Cancer were used for the statistical analysis of data. Weekly palpations were made along the course of studies and general parameters were monitored. The only effects attributed to ebrotidine administration were a slight decrease in the survival rate of female mice given the 500 mg/kg dose and a lower weight gain in rats of both sexes. The histopathological data revealed that lipoid pneumonia and kidney calculi are more frequent in rats treated with doses of 500 and 300 mg/kg. No increment in the spontaneous occurrence of tumours or significant presence of tumours in treated animals differing from that in control animals was observed, and a decrease in the time required for their onset that could be related to ebrotidine was not observed either. There were no differences in hyperplastic and/or dysplastic changes between treated and control animals. Therefore, it is deduced that ebrotidine does not induce neoplastic or preneoplastic effects in rats or mice even at doses of 500 mg/kg, at which some general toxicity effects are seen. PMID- 9205756 TI - Study of the population of antral G-cells and enterochromaffin-like cells in the rat and mouse gastric mucosa after long-term treatment with ebrotidine. AB - The potential effect of ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene]amino]-4 thiazolyl]methyl]thio]ethyl ] amino)methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) on two types of enteroendocrine cell populations in the gastric mucosa, antral G-cells and enterochromaffin-like cells, was investigated. The study of the population of antral G-cells was performed in a group of male rats treated with ebrotidine 500 mg/kg p.o. for 60 days; a control group receiving 10 ml/kg of an aqueous agar solution was used. A PAP (peroxidase antiperoxidase) system-associated antigastrin immunohistochemical method was used for cell identification. The population of enterochromaffin-like cells was assessed by quantifying the density of argyrophilic cells in mouse gastric mucosa after an 18-month treatment with ebrotidine 500 mg/kg. Grimelius silver staining method was used for cell identification. In both studies, cell count was performed using a light microscope at 400 x magnification and cell density was calculated by computer-assisted image analysis. Compared to control, ebrotidine did not cause any significant differences in the cell density of the populations studied. PMID- 9205757 TI - Comparative study of plasma gastrin levels in rats after two months of ebrotidine administration. AB - Four groups of male rats were orally administered for 60 days with daily doses of ebrotidine (N-[(E)-[[2-[[[2-[(diaminoethylene) amino]-4 thiazolyl]methyl]thio]ethyl]amino]methylene]-4-bromo- benzenesulfonamide, CAS 100981-43-9, FI-3542) (500 mg/kg), ranitidine (500 mg/kg), cimetidine (500 mg/kg) and omeprazole (43.5 mg/kg). A fifth group received no treatment and was used as control. The curve of gastrinemia was obtained on days 1, 15 and 60 of administration. On each of these days gastrinemia was assessed at 0, 1, 5, 8, and 24 h on day 1, and 1, 5, 8, 10 and 24 h on days 15 and 60. The purpose of this study was to compare the plasma gastrin level profile in association with the administration of test drugs on days 1, 15 and 60 of treatment. The results showed a significant difference in the duration of hypergastrinemia of H2 receptor antagonists as compared to proton pump blockers. Although peak plasma gastrin levels were attained for all products between 5 and 8 h after day 1 of administration, H2-receptor antagonists, unlike omeprazole, achieved recovery of gastrin baseline levels within 24 h. On days 15 and 60 of ebrotidine, treatment, plasma gastrin levels returned to normal range at 5 and 8 h after administration, respectively. After ranitidine and cimetidine, hypergastrinemia was still present at this time, but normal levels were attained before 24 h. With omeprazole plasma gastrin levels did not return to normal range within 24 h after each administration, and a cumulative effect occurred during treatment. The omeprazole treated group showed the highest and more sustained plasma gastrin levels. It was concluded that ebrotidine was the antisecretory agent with the lowest hypergastrinemic effect during long-term treatment. With ebrotidine daily baseline gastrin levels were more rapidly recovered after each administration. PMID- 9205758 TI - Tolerability and pharmacokinetics of ebrotidine in healthy subjects given single and repeated oral doses. AB - The tolerability and safety of ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl]methyl] thio]ethyl]amino]methylene]-4-bromo benzenesulfonamide, CAS 100981-43-9, FI-3542) and its basic pharmacokinetic parameters were determined after its oral administration to healthy volunteers. Sixteen subjects were selected to participate in two different studies: an increasing single dose study to determine the maximal tolerated dose (from 25 to 1600 mg), and a multiple dose study (stepped doses from 400 to 1600 mg daily for 12 days). The results of the studies showed that ebrotidine has a good tolerability. Vital signs and laboratory tests were not influenced by the study treatment. No clinically relevant adverse effects were reported during the investigation. Ebrotidine reached peak plasma concentrations 2-3 h after oral administration. Its elimination half-life ranged from 9 to 14 h. In conclusion, ebrotidine was well tolerated after administration of oral single doses of up to 1600 mg, and after repeated administration of up to 800 mg/12 h for 12 days. PMID- 9205760 TI - Pharmacokinetics of ebrotidine in healthy volunteers. A summary. AB - Several clinical pharmacokinetic studies of ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl] methyl]thio]ethyl]amino]methylene]-4-bromo benzenesulfonamide, CAS 100981-43-9, FI-3542) administered by oral route in single and multiple doses to healthy volunteers have been performed. Dosage levels were 150, 300, 400, 500, 600 and 800 mg. Plasma concentrations of unchanged ebrotidine and its major metabolite, ebrotidine sulfoxide, excreted in the urine were determined. The main pharmacokinetic parameters were calculated from the experimental data. Absorption was relatively rapid (Imax = 2 h) and unrelated to dose. Drug behavior was considered as reasonably linear: Cmax = 364 1168 ng/ml and AUC0-12 h = 1427-5997 ng.h/ml (doses from 150 mg to 800 mg). The mean values of terminal elimination half-life (t1/2 beta) ranged from 13.9 to 20.3 h (doses of 400, 600 and 800 mg). After multiple dosing there was no drug accumulation, and no significant changes in the mean values of the main pharmacokinetic parameters were observed. The steady state was reached from the second day of administration, 10-24% of the ebrotidine administered dose was excreted in urine mainly as its major metabolite, ebrotidine sulfoxide, as well as unchanged drug and other minor metabolites. These percentages were constant and independent of the dose administered. PMID- 9205759 TI - Pharmacokinetic study of ebrotidine administered in multiple doses to healthy volunteers for 4 days. AB - The safety of ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]-4- thiazolyl]methyl]thio]ethyl]amino] methylene]-4-bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542), a new H2-receptor antagonist with gastroprotective activity, was assessed and its main pharmacokinetic parameters were determined in order to establish the dose linearity after the repeated administration of three different dose levels. The study was carried out in a group of 8 healthy volunteers of either sex, aged between 20 to 29 years. Oral doses of ebrotidine were administered in a randomized, single-blind design. Volunteers remained in the Unit for two days at each of the three study phases with washout intervals of 2 weeks and received seven doses of ebrotidine (150, 300 and 500 mg b.i.d). Pharmacological evaluation included vital signs, laboratory tests, adverse events and blood and urine samplings for pharmacokinetic analysis. Ebrotidine was determined by high performance liquid chromatography (HPLC) with UV detection. The results showed a good tolerability of ebrotidine after the administration of seven doses for 4 days, with no changes in the vital signs or laboratory parameters. No clinically significant dose-related adverse events were reported during the study. The absorption of ebrotidine was relatively rapid (tmax approximately 2 h) and linear within the dose range from 150 to 500 mg. Drug biotransformation was linear with doses tested, and no metabolic saturation occurred. The terminal elimination half-life of ebrotidine was between 7 and 11 h or even longer. There was no accumulation of ebrotidine and the steady state was reached, regardless of the dose administered, within the first 24-48 h. PMID- 9205761 TI - Continuous intragastric pH monitoring in the evaluation of ebrotidine, cimetidine and placebo on gastric acidity in healthy volunteers. AB - This study was conducted to determine the efficacy and tolerance of ebrotidine (N [(E)-[[2-[[[2-[(diaminomethylene)amino]-4-thiazolyl) methyl]thio]ethyl]amino]methylene]-4-bromo-benzenesulfonamide, CAS 100981-43.9, FI-3542), a new H2-receptor antagonist, on reducing gastric acidity after a single 800 mg dose, compared with cimetidine 800 mg once daily and placebo by means of a continuous intragastric pH monitoring. A total of 30 healthy volunteers were allocated to receive in a double blind, parallel design the study medication. Clinical observations, physical examinations and visual analogue scales (VAS) were performed during the study to assess the tolerability of the three treatments. Ebrotidine and cimetidine caused a greater and longer-lasting gastric acid inhibition than placebo. With ebrotidine, significantly (p < 0.05) higher median pH values (and interquartile range, IQR) were reached in the post administration (2.61, IQR 2.02-3.93), postprandial (3.38, IQR 2.82-3.91) and nocturnal (2.83, IQR 1.69-3.77) periods than with placebo: 1.82 (IQR, 1.66-2.09), 2.81 (IQR, 2.02-3.28), and 1.89 (IQR, 1.44-2.13), respectively. Cimetidine showed significant differences compared to placebo in the post-administration (2.36, IQR 1.89-3.46) and nocturnal (2.46, IQR 1.88-4.33) periods. No statistical differences were observed between the active treatments. Ebrotidine caused a significantly higher percentage of time above pH 2.0 in the post-administration and nocturnal periods compared to placebo (p < 0.05), and above pH 3.0 in the post-administration, postprandial and nocturnal periods. No serious adverse effects, or disturbances in the VAS or in the vital signs were reported, and all medications were well tolerated. It is concluded that a single dose of ebrotidine 800 mg is as effective as cimetidine 800 mg in reducing total and nocturnal intragastric acidity. The study also confirms the excellent safety profile of the new drug. PMID- 9205762 TI - Comparison of the efficacy and safety of ebrotidine in the treatment of duodenal ulcer. A multicentre, double-blind, placebo-controlled phase II study. AB - Ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]-4 thiazoly]methyl]thio]ethyl]amino]methylene]-4-bromo-benzenesulfon amide, CAS 100981-43-9, FI-3542) is a new H2-receptor antagonist characterized by its high receptor affinity and gastroprotective effect. This Phase II study has been undertaken to establish the efficacy and safety of ebrotidine, administered in four dosages as a single evening dose versus placebo in the treatment of duodenal ulcer. A total of 110 duodenal ulcer patients were studied in a randomized, double-blind, placebo-controlled, multicentre clinical trial. The patients were assigned to 5 groups: placebo, 200 mg, 400 mg, 600 mg and 800 mg of ebrotidine once daily. Controls were performed at baseline and every two weeks at four follow-up visits unless ulcer healed before. Endoscopic examination was the main parameter for the assessment of treatment efficacy and ulcer healing rate. Vital signs and blood/ urine analysis were used to establish safety. The three groups treated with higher dosages (400 to 800 mg of ebrotidine daily) showed an endoscopic ulcer healing rate of 90-95%, significantly higher than 55% achieved with placebo (p < 0.05), whilst the differences between these three dosages of ebrotidine were not statistically significant. Healing rate in the group treated with 200 mg of ebrotidine daily was not significantly different from that in the placebo group. The development of symptoms, number of episodes of ulcer-related pain, total ulcerated surface area or subjective ratings by the patients and investigators also differed significantly between ebrotidine (400, 600 and 800 mg daily) and placebo, and again, no marked differences were found between these three doses of ebrotidine. As far as tolerance is concerned, no clinically or statistically significant changes were observed in vital signs and analytical parameters. The incidence of side effects was less than that presented by the placebo group, possibly due to a greater consumption of antacids in this group. Results showed that a daily dose of 400 mg ebrotidine is effective and safe in the treatment of duodenal ulcers. PMID- 9205763 TI - Ebrotidine versus ranitidine in the healing and prevention of relapse of duodenal ulcer. A multicentre, double-blind, parallel, randomized, controlled study. AB - Two hundred and fifty patients were included in a double-blind, parallel, randomized, controlled clinical trial. Duodenal ulcer treatment lasted up to 8 weeks. Forty-nine patients were followed up for prevention of ulcer relapse for up to one year. All patients received either ranitidine (300 mg/day in the healing phase and 150 mg/day in the follow-up phase) or ebrotidine (N-[(E)-[[2 [[[2-[(diaminomethylene)amino]-4 -thiazolyl]methyl]thio]ethyl]amino]methylene]-4 bromo-benzenesulfonamide , CAS 100981-43-9, FI-3542) (400 mg/day in both phases) as a single dose at bedtime. Both groups were matched in all demographic parameters, except for a significantly higher percentage of smokers in the ranitidine group. The percentage of total healing was almost the same with both products. Healing occurred in a higher percentage with ebrotidine at weeks 4 (75% versus 66.7%) and 6 (87% versus 79.7%). A higher effect of ebrotidine on the incidence of duodenitis was identified during the whole study, but only reached statistical significance at week 6. The relapse rate during the follow-up phase showed no differences between the two study treatments, relapse percentage figures being 25% for ebrotidine and 24% for ranitidine. There were no differences in the number of unscheduled visits between the two groups, although 57% of patients in the ranitidine group had to make a second follow-up visit, as compared with 33% in the ebrotidine group. Both drugs caused hardly any side effects, affecting only one patient from each group: one patient with ebrotidine suffered from diarrhoea and one patient with ranitidine developed a skin rash on the limbs. Administration of ebrotidine in a single dose (400 mg/d) was at least as effective and safe as ranitidine both for healing and relapse prevention in patients with duodenal ulcer. PMID- 9205764 TI - Ebrotidine versus ranitidine in the treatment of acute duodenal ulcer. A multicentre, randomized, double-blind, controlled clinical trial. AB - A total of 478 patients with endoscopically confirmed duodenal ulcer entered this randomized, parallel, double-blind trial. Patients were randomly assigned to receive ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]- 4 thiazolyl]methyl]thio]ethyl]amino]methylene]-4-bromo-benzenesulfona mid e, CAS 100981-43-9, FI-3542) 400 mg or ranitidine 300 mg tablets (4:1) respectively, administered in single evening doses. Endoscopy, clinical examination and symptom assessment were performed at baseline and at weeks 4 and 8. Safety evaluations including laboratory tests, treatment compliance and antacid consumption checks were conducted at the beginning and/or at the 4 and 8 week visits. Patients whose ulcer showed endoscopic healing at the 4-week control left the study. Both groups were matched in all parameters studied. The healing rates at 4 weeks were 76.4% and 75.3% for ebrotidine and ranitidine respectively, while at 8 weeks the final rates were 95% and 91.8% respectively. Accompanying symptoms disappeared rapidly and the patients returned to normal. Smoking proved to be a highly significant negative risk factor, since healing rates were 83.4% and 71.2% at 4 weeks and 97.4% and 92.3% at 8 weeks in non-smokers and smokers respectively (p = 0.0046). Smokers treated with ranitidine showed significantly lower final healing rates than non-smokers (86% vs 100%; p = 0.0358), while the healing rates among patients treated with ebrotidine were similar regardless of whether they were smokers or not (93.9% and 96.7% N.S.). Ebrotidine (94%) proved to be more effective than ranitidine (86%) in smokers with higher healing rates (p < 0.05). Alcohol intake showed no significant relationship with the healing rates. Both drugs demonstrated an excellent safety. There were no changes in blood parameters, and no significant adverse events were reported. PMID- 9205765 TI - Efficacy of ebrotidine and ranitidine in the treatment of benign gastric ulcer. AB - This is a phase III, randomized, double-blind, clinical trial with two parallel groups of 50 patients to assess the efficacy of ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl]methyl]thio]ethyl ] amino]methylene]-4 bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) 800 mg and ranitidine 300 mg as a single evening dose in the treatment of benign gastric peptic ulcer. Prior to treatment, an endoscopy was performed to detect ulcer lesions and to discard malignancies. Clinical and endoscopic examinations were performed at 6, 9 and 12 weeks. Healing rates were significant for both treatments at week 6, while at week 12 there was statistical significance for ebrotidine as compared to ranitidine (96% vs 88% in the intention-to-treat analysis and 98% vs 87.5% in the per protocol analysis). Decrease in ulcer diameter was significant for both treatments at week 6, and for ebrotidine versus ranitidine at weeks 9 and 12. The overall improvement of symptoms was higher with ebrotidine, which was already significant at week 6. Safety was considered to be excellent, since no significant adverse events were reported for the patients included in the study. PMID- 9205766 TI - Studies on the protective effect of ebrotidine on experimental ulcers induced by non-steroidal anti-inflammatory drugs in healthy volunteers. AB - Ebrotidine (N-[(E)-[[2-[[[2-[(diaminomethylene)amino]- 4 thiazolyl]methyl]thio]ethyl]amino]methylene]-4-bromo-benzenesulfonamid e, CAS 100981-43-9, FI-3542) is a new H2-receptor antagonist providing a new therapy for the prevention and healing of non-steroidal anti-inflammatory drugs-induced gastroduodenal lesions. Carbonic anhydrase is a zinc enzyme, and its isozyme (carbonic anhydrase II) in parietal cells plays a central role in HCl secretion. The effects of ebrotidine on carbonic anhydrase in human subjects are reported. Eighteen healthy volunteers were distributed in 3 equal subgroups and treated for 10 days as follows: ebrotidine 800 mg/d p.o. (Group A); indometacin 4 mg/kg/d p.o. in 3 divided doses (Group B); ebrotidine 800 mg/d p.o. plus indometacin 4 mg/kg/d p.o. (Group C). Assessment of the enzymatic activity of carbonic anhydrase was based on the colorimetric method of changing pH with the stopped flow technique. In group A, ebrotidine reduced total gastric mucosal carbonic anhydrase activity by 62%; in group B, indometacin increased carbonic anhydrase activity in gastric mucosa by 138%; in group C, the combined treatment with ebrotidine plus indometacin decreased gastric mucosal carbonic anhydrase activity by 38%. The present study shows that, unlike ranitidine, ebrotidine, a competitive H2-receptor antagonist, is also a non-competitive inhibitor of carbonic anhydrase I and II. By antagonizing the activating effects of indometacin on gastric mucosal carbonic anhydrase, ebrotidine prevents mucosal lesions caused by anti-inflammatory drugs. PMID- 9205767 TI - Comparative study of the safety and efficacy of ebrotidine versus ranitidine and placebo in the prevention of piroxicam-induced gastroduodenal lesions. AB - This study assessed the efficacy of ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene)amino]-4-thiazolyl] methyl]thio]ethyl]amino]methylene]-4-bromo benzenesulfonamide, CAS 100981-43-9, FI-3542) versus ranitidine and placebo in preventing gastroduodenal lesions induced by piroxicam. Thirty patients with rheumatic disease, who were divided into 5 groups, received an oral treatment of piroxicam 20 mg once daily for 6 days plus ebrotidine 400 mg/day (Group I); ebrotidine 800 mg/day (Group II); ranitidine 150 mg/day (Group III); ranitidine 300 mg/day (Group IV); or placebo (Group V). Patients were endoscopically examined before and after treatment. Lanza's score was also determined, and laboratory tests were performed. The results of this study showed that the most powerful protective effect against mucosal gastric lesions induced by piroxicam was achieved with 800 mg/day of ebrotidine. Ranitidine at doses of 150 mg/day did not protect gastric mucosa, and the 300 mg/day dose exerted a poor gastroprotective effect. PMID- 9205768 TI - Efficacy of ebrotidine and ranitidine combined with amoxicillin and metronidazole in the eradication of Helicobacter pylori in patients with duodenal ulcer. AB - This double-blind, randomized, phase III clinical trial was carried out in two parallel groups to assess the efficacy of ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene) amino]-4-thiazolyl]methyl]thio]ethyl]amino] methylene]-4 bromo-benzenesulfonamide, CAS 100981-43-9, FI-3542) 400 mg and ranitidine 300 mg given in single evening dose, combined with amoxicillin 750 mg and metronidazole 500 mg three times daily for 14 days, in the eradication of Helicobacter pylori in patients with duodenal ulcer. Thirty patients were included, divided into two groups of 15, to whom one of the study therapies was administered based on a randomization code. Clinical and endoscopic controls were performed 4, 6 and 8 weeks after the onset of the treatment. No differences were seen between the two treatment groups with regard to demographic parameters and clinical histories. They were both perfectly homogeneous. There were no differences between the eradication of both therapies in both the antrum and gastric body samples (over 80% eradication), allowing the results to be classified as satisfactory. Moreover, perfect control was achieved through the study of clinical symptoms, which even disappeared in some cases. There were no differences in the healing rate of the duodenal ulcer after four weeks, 86.7% being achieved for both groups. PMID- 9205770 TI - De novo analysis of receptor binding affinity data of 8-ethenyl-xanthine antagonists to adenosine A1 and A2a receptors. AB - The receptor binding affinity data to adenosine A1 and A2a receptors of a wide series of 8-ethenyl-xanthine derivatives has been analyzed by means of the Free Wilson model. The analysis of the individual group contributions (aij) shows the importance of the presence of an ethenyl moiety at position 8 on the xanthine ring for obtaining selective A2a antagonists. The different aij values of the substituents for the adenosine. A1 receptor do not correlate with the corresponding ones for the A2a receptor, indicating the possibility to obtain A1 and A2a selective compounds. The presence of aromatic substituents at the 8 ethenyl group, such as 3,5-(OCH3)2-phenyl, permits to obtain strongly A2a selective compounds (affinity ratio of up to 100); moreover, it appears that 8 ethenyl-xanthinic derivatives cannot have high selectivity for the adenosine A1 receptor (affinity ratio < or = 10). PMID- 9205769 TI - Studies on the cytoprotective and antisecretory activity of ebrotidine. A review. AB - Gastric mucosa is exposed to various aggressive factors such as stress, ulcerogenic drugs including acetyl-salicylic acid(ASA)-like agents, ethanol, bacteria, particularly Helicobacter pylori (Hp), and various endogenous irritants such as acid-pepsin secretion and bile salts. The maintenance of the mucosal barrier depends upon the activation of the pre-epithelial (mucus-alkali secretion), epithelial (surface-active phospholipids and rapid mucosal restitution) and post-epithelial (mucosal microcirculation, sensory nerves and mast cells) components of mucosal defense. Ebrotidine (N-[(E)-[[2-[[[2 [(diaminomethylene)amino]- 4-thiazolyl]methyl]thio]ethyl]amino]methylene]-4-bromo benzenesulfonamid e, CAS 100981-43-9, FI-3542) is the first of a new generation of H2-receptor antagonists with both antisecretory and cytoprotective activities. Its inhibitory action is similar to that of ranitidine and approximately tenfold greater than cimetidine, and is accompanied by a small and transient increase in plasma gastrin levels. In contrast to ranitidine and other H2-receptor antagonists, ebrotidine exerts a unique cytoprotection against injury by various ulcerogens such as ethanol, ammonia, lipopolysaccharides (LPS), stress and ASA or acidified taurocholate. The mechanism of this protection by ebrotidine is not clear, but it has been shown to stimulate mucus secretion, to increase the quality of adherent mucus gel and to increase gastric mucosal blood flow (GBF), possibly due to enhanced mucosal formation of prostaglandin E2 (PGE2) and nitric oxide (NO). The cytoprotective effects of ebrotidine were observed in rats and confirmed also in humans with gastric lesions induced by ethanol or ASA. Ebrotidine also exerts anti-Helicobacter pylori (Hp) effects by interfering with surface receptors of epithelial cells and inhibiting urease, protease and lipase activity, and by counteracting the noxious effects of Hp-related substances such as ammonia and lipopoly-saccharides (LPS). PMID- 9205771 TI - Suppression of lipopolysaccharide-induced impairment of active avoidance and interleukin-6-induced increase of prostaglandin E2 release in rats by indometacin. AB - The effects of indometacin (CAS 53-86-1) on lipopolysaccharide-induced impairment of active avoidance and on interleukin-6-induced increase of prostaglandin E2 release were investigated in rats. In the experiment on acquisition and retention of one-way active avoidance in a shuttle box model, bilateral infusion of lipopolysaccharides (LPS) into the hippocampus, 1 microgram per side, resulted in a significant impairment both in acquisition and retention by prolonging the latency of avoidance in training and testing. In the meantime, intraperitoneal injection of indometacin 10 mg/kg daily for 7 days, improved the LPS-induced amnesia especially in the testing by shortening the latency from 2.3 to 1.7 s (p < 0.05). In the in vivo microdialysis study in anesthetized rats, intrahippocampal infusion of 80 ng interleukin-6 (IL-6) markedly increased prostaglandin E2 (PGE2) release into hippocampal dialysates which started at 2 h post administration. Perfusion of indometacin (0.3 mol/l) into the hippocampus for 1 h obviously suppressed the IL-6-induced PGE2 response. These findings provide experimental evidence that--assuming that central inflammation may be involved with Alzheimer's disease a non-steroidal anti-inflammatory drug may be used in the treatment of Alzheimer's disease. PMID- 9205772 TI - Human bioequivalence study of a new nifedipine containing retard filmtablet after single and repeated administration. AB - A clinical pharmacokinetic bioequivalence study with two retard filmtablet preparations, both containing 20 mg of nifedipine (CAS 219829-25-4) was carried out. The investigated test preparation was Cordaflex 20 mg retard filmtablet. The pharmacokinetic parameters were determined after single and repeated administration in 15 and 16 healthy male volunteers, respectively, in open, randomised studies of cross-over design. Plasma levels of nifedipine were determined by HPLC with electrochemical detection using a robotic sample preparation technique. Statistical comparison of the pharmacokinetic parameters (AUC0-infinity, AUCss, tau tmax, Cmax, Css,min, Css,av, MRT, etc.) calculated from plasma concentration-time curves by ANOVAlog, confidence interval, Schuirman's, Westlake's, Anderson's and Wilcoxon's tests, furthermore the comparison of the clinical results did not show any significant difference between the two preparations. It is concluded that the two preparations are bioequivalent after repeated administration. PMID- 9205773 TI - Pharmacological profile of valsartan, a non-peptide angiotensin II type 1 receptor antagonist. 1st communication: antihypertensive effects of valsartan in hypertensive models. AB - The antihypertensive effects of valsartan ((S)-N-valeryl-N-?[2'-(1H-tetrazol-5 yl)biphenyl-4-yl]methyl? valine, CAS 137862-53-4, CGP 48933), an angiotensin II type 1 receptor antagonist, were examined in hypertensive rats and dogs. In normotensive rats and deoxycorticosterone acetate (DOCA)/salt hypertensive rats, valsartan had no effect on blood pressure. Single oral administrations of valsartan at doses of 3-30 mg/kg reduced blood pressure dose-dependently in renal (2 kidney 1 clip, 2K1C) hypertensive and spontaneously hypertensive rats (SHR). Repeated oral administrations of valsartan to these hypertensive rats controlled blood pressure throughout a treatment period of 4 weeks, and showed no rebound phenomenon following drug withdrawal. This drug at 30 mg/kg p.o. decreased blood pressure in renal (2K1C) hypertensive dogs by single and repeated administrations. The extent and duration of the hypotensive action of valsartan were similar to those of enalapril. Valsartan would thus appear as useful as enalapril. PMID- 9205774 TI - Pharmacological profile of valsartan, a non-peptide angiotensin II type 1 receptor antagonist. 2nd communication: valsartan prevents end-organ damage in spontaneously hypertensive stroke-prone rats during 1-year treatment. AB - Valsartan ((S)-N-valeryl-N-?[2'-(1H-tetrazol-5-yl)biphenyl-4-yl]methyl? valine, CAS 137862-53-4, CGP 48933), a non-peptide angiotensin II type 1 receptor antagonist, or enalapril was administered to spontaneously hypertensive rats stroke-prone (SHR-SP) for 1 year from 8 weeks to 56 weeks of age under a normal diet without saline load. During 48 weeks, control rats showed increase in systolic blood pressure from 180 to 250 mmHg accompanying stroke-related behaviour, cardiac and aortic hypertrophy, hyperreactive contractility of mesenteric vascular beds, proteinuria, high water turnover and death. Valsartan at 3, 10 and 30 mg/kg/d p.o. and enalapril at 1 and 10 mg/kg/d p.o suppressed the increase in blood pressure dose-dependently. Systolic blood pressure was steadily controlled to around 180 mmHg at the highest dose of either drug throughout the study. In proportion to the antihypertensive action of the drugs, end-organ damage was prevented. During 1-year administration, effects of enalapril and valsartan were much the same, indicating the clinical usefulness of valsartan being comparable to enalapril. PMID- 9205775 TI - Pharmacological profile of valsartan, a non-peptide angiotensin II type 1 receptor antagonist. 3rd communication: hemodynamic effects of valsartan in rats and dogs. AB - Valsartan ((S)-N-valeryl-N-?[2'-(1H-tetrazol-5-yl)biphenyl-4-yl)methyl? valine, CAS 137862-53-4, CGP 48933) at 10 and 30 mg/kg p.o. significantly inhibited angiotensin I and angiotensin II-induced pressor response at 1-8 h after administration in normotensive conscious dogs, but had no effect on blood pressure, heart rate or cardiac functions such as cardiac output, left ventricular pressure, left ventricular max. dP/dt and left ventricular end diastolic pressure. In contrast to enalapril, this drug did not potentiate bradykinin-induced depressor response. In anesthetized dogs, valsartan at 10 mg/kg i.v. significantly increased renal blood flow, urinary sodium and chloride excretion and urine volume and significantly decreased renal vascular resistance and filtration fraction without affecting blood pressure. The preferential effects on renal hemodynamics were confirmed by the increase in blood flow into the kidneys of spontaneously hypertensive rats at 3 h following oral administration. PMID- 9205776 TI - Pharmacological profile of valsartan, a non-peptide angiotensin II type 1 receptor antagonist. 4th communication: improvement of heart failure of rats with myocardial infarction by valasartan. AB - The hemodynamic effects of valsartan ((S)-N-valeryl-N-?[2'-(1H-tetrazol-5 yl)bipheneoyl-4-yl]meth yl?valine, CAS 137862-53-4, CGP 48933), a new angiotensin II type 1 receptor antagonist, on rats with myocardial infarction induced by coronary artery ligation was examined. Four weeks after ligation, mean blood pressure, left ventricular pressure and cardiac output decreased, while left ventricular end-diastolic pressure increased in control rats. Left ventricular end-diastolic pressure significantly decreased in rats treated with valsartan at 30 mg/kg/d p.o. for 4 weeks. Total systemic resistance remarkably decreased in those with enalapril 3 mg/kg/d p.o. and valsartan 30 mg/kg/d p.o. Valsartan and enalaprilat did not affect cardiac functions of isolated intact rat hearts before and after ischemia in Langendorff apparatus. In addition to hemodynamic effects observed in vivo, valsartan at 30 mg/kg p.o. significantly inhibited left ventricular hypertrophy. Valsartan would thus appear to be clinically useful for treating heart failure following myocardial infarction. PMID- 9205777 TI - Pharmacological profile of valsartan, a non-peptide angiotensin II type 1 receptor antagonist. 5th communication: hemodynamic effects of valsartan in dog heart failure models. AB - Hemodynamic effects of valsartan ((S)-N-valeryl-N-?[2'-(1H-tetrazol-5-yl)biphenyl 4-yl]meth yl?valine, CAS 137862-53-4, CGP 48933), a non-peptide angiotensin II type 1 receptor antagonist were examined in dogs with heart failure induced acutely by coronary artery ligation and chronically by rapid-ventricular pacing. Coronary artery ligation induced decrease in cardiac output and increase in left ventricular end-diastolic pressure. Valsartan at 10 mg/kg i.v. reduced blood pressure, heart rate, left ventricular pressure, left ventricular end-diastolic pressure and total systemic resistance. Similar changes were observed with enalaprilat at 0.1 mg/kg i.v. Rapid left ventricular pacing for 2 weeks reduced cardiac contractility. Valsartan, administered at a dose of 100 mg/kg/d p.o. for 2 weeks, lowered left ventricular end-diastolic pressure. Valsartan reduced preload and afterload in these two dog heart failure models. PMID- 9205778 TI - Synthesis and anti-inflammatory activity of polyazaheterocyclic derivatives of 6 amino-2,4-lutidine and their precursors. AB - Derivatives of N-(4,6-pyridin-2-yl)arylcarboxamides resulting from the integration of the amidic function into 4H-1,2,4-triazole, triazol-3(2H)-one and 1H-tetrazole rings were evaluated as potential anti-inflammatory compounds. The level of activity decreased as compared to carboxamides, nevertheless their precursors and notably the corresponding amidrazones exhibited potent activity; amidrazone 21, whose ID50 was 34.4 mg.kg-1 in the rat paw edema test, was selected for further investigation. These heteroarylcarboxamide derivatives could represent an interesting alternative to classical non-steroidal anti inflammatories in so far as they partly act by inhibition of tumor necrosis factor-alpha production. PMID- 9205779 TI - Synthesis, antioxidant and anti-inflammatory activity of novel substituted ethylenediamines and ethanolamines. A preliminary quantitative structure-activity relationship study. AB - Six substituted oxo- or hydroxy-aminoethanols and ethylenediamines were synthesized and tested as anti-inflammatory agents. 1-Substituted 4-(2 aminoethylamino)-1-butanones and 1-substituted 4-(2-hydroxy-ethylamino)-1 butanones were prepared by reacting the appropriate 4-chloro-1-butanone with the corresponding aminoalcohol or ethylenediamine. 1-Substituted 4-(2 aminoethylamino)-1-butanols were prepared by the reduction of the ketones with NaBH4 or NaBH3CN. The RM values of the synthesized compounds were determined as an expression of their lipophilicity. The effect of these compounds on in vitro non-enzymatic lipid peroxidation, their hydroxyl radical scavenging activity and their ability to interact with 1,1-diphenyl-2-picrylhydrazyl stable free radical (DPPH) were studied. The effect of the synthesized compounds on inflammation, using the carrageenan induced rat paw edema model was studied. Both anti inflammatory and antioxidant activities depended on some structural characteristics of the synthesized compounds. It was also attempted to correlate the above mentioned activities with some physicochemical parameters using a quantitative structure-activity relationship approach. The primary amino group appeared to be of importance for antioxidant activity in this series of compounds. PMID- 9205780 TI - Pharmacological profile of the novel potent antirheumatic 4-(2',4' difluorobiphenyl-4-yl)-2-methyl-4-oxobutanoic acid. AB - On the basis of basic screening for novel, more potent antiarthritics VUFB-16066 (4-(2',4'-difluorobiphenyl-4-yl)-2-methyl-4-oxobutanoic acid, CAS 112344-S2-2) was chosen as a compound with pronounced anti-inflammatory and immunomodulatory effects, with good gastric tolerance and relatively low toxicity. VUFB-16066 is a dual cyclooxygenase and 5-lipoxygenase inhibitor, and it suppresses alloantigen driven cellular immune response and phagocytosis of stimulated peritoneal cells. VUFB-16066 exhibits prolonged pharmacological activity connected with its major metabolite having a very long half-life. In the model of adjuvant arthritis VUFB 16066 improves most of disease symptoms including immunopathological disturbances, which indicates possible disease-modifying activity of the drug. The beneficial antiarthritic effect of VUFB-16066 has been also confirmed in patients with rheumatoid arthritis. PMID- 9205781 TI - Enantioselective biotransformation of the chiral antihistaminic drug dimethindene in humans and rats. AB - The biotransformation of dimethindene (CAS 5636-83-9, dimethindene maleate, CAS 3614-69-5) after oral administration was determined in urine using an HPLC gradient method. Besides the phase I metabolites the conjugates of the hydroxylated metabolites with glucuronic and/or sulfuric acid were quantitatively determined after enzymatic deconjugation. The cumulative excretion of 6 hydroxydimethindene and 6-hydroxy-N-demethyldimethindene in human urine after hydrolysis of the conjugates ranged from 18 to 23% of the administered dose, independent of the amount of the dose applied. The results indicate that conjugated 6-hydroxydimethindene is the main metabolite of dimethindene. Increasing doses of 5 to 20 mg dimethindene maleate did not affect the relative amount of the excreted metabolites but changed the ratio of 6-hydroxydimethindene to 6-hydroxy-N-demethyldimethindene from 3:1 to 1:1. In rats about 4 to 8% of the administered dose of dimethindene was excreted as dimethindene-N-oxide which is the main metabolite in rat urine. After administration of R-(-)-dimethindene the elimination of all metabolites was 2 to 3 fold higher compared to the administration of the S-(+)-enantiomer. By chiral HPLC, in 10 human volunteers a stereoselective elimination of N-demethyl-dimethindene after oral administration of racemic dimethindene with the predominant excretion of the R-(-)-enantiomer was observed. PMID- 9205782 TI - Studies on the effect of ursodesoxycholic acid on rats with acute carbontetrachloride injury. AB - Bilirubin is a sensitive marker of toxic liver injury. Carbontetrachloride (CCl4) is used in some manufactures and laboratories. An acute intoxication is a peril. We have performed experiments to state total, indirect and direct bilirubin levels in rats exposed to a single CCl4 dose of 1.25 ml/kg. Total bilirubin in normal rats is 3.13 +/- 0.13 mumol/l. 58.6% is direct conjugated and 41.3% indirect unconjugated bilirubin. One hour after CCl4 administration bilirubin started to increase and reached its peak in the second hour. 24 h later a decrease began but even 72 h after intoxication it was not normalized. Ursodesoxycholic acid (UDCA, CAS 128-13-2, Ursofalk) treatment (25 mg/kg/dose) lowered bilirubin, returning to normal level 24 h after CCl4 administration. Hypothermia aggravates intoxications. In CCl4 lesion it develops 1 h after exposure. UDCA has not influenced low body temperature but liver function, conjugating capacity was restored. In CCl4 poisoning it is suggested to start UDCA treatment as early as possible and to continue it for some more days after improvement of serum bilirubin concentration. PMID- 9205783 TI - Synthesis and antimycobacterial activity of nitroxanthones. 1st communication: synthesis and differential scanning calorimetry analysis. AB - The syntheses of mono-; di-, tri- and tetranitroxanthones via intramolecular Friedel-Crafts acylation of nitrated 2-phenoxybenzoic acids as well as via selective stepwise nitration of xanthone and the nitroxanthones gained from the first route are described. The synthesized nitroxanthones investigated by DSC (differential scanning calorimetry) analysis show a surprisingly high thermodynamic stability which was confirmed by quantum chemical calculations. PMID- 9205784 TI - Antifungal activity of organic and organometallic derivatives of benzimidazole and benzothiazole. AB - Forty organic or organometallic derivatives of benzimidazole and benzothiazole and five rhodium(I) and ruthenium(II) complexes were evaluated for their in vitro antifungal activity against Candida albicans. Four of the tested compounds, the rhodium containing compounds 30, 31, 32 and 33, were found effective at the minimum inhibitory concentrations(MICs) between 400-600 micrograms/ml. PMID- 9205785 TI - Activity of cytotoxin P4 from the venom of the cobra snake Naja nigricollis on gram-positive bacteria and eukaryotic cell lines. AB - The cytotoxin P4 from the venom of the cobra snake Naja nigricollis was examined for activity against aerobic gram-positive and gram-negative bacteria, yeasts, fungi and eucaryotic cell lines by determination of minimal inhibitory concentrations (MIC) and IC50 values, respectively. Cytotoxin P4 exhibits antimicrobial activity against Bacillus subtilis, Micrococcus flavus and Sarcina lutea. Targets in gram-negative bacteria and fungi apparently were not reached. Toxicity against eucaryotic cells is in a narrow range between lethal and tolerable concentrations. PMID- 9205786 TI - Cefodizime once daily in the treatment of lower respiratory tract infections. AB - Cefodizime (CAS 69739-16-8, HR 221) is a new third-generation cephalosporin with pharmacokinetic properties that make it suitable for once-daily administration in the treatment of lower respiratory tract infections (LRTI). Ninety-nine adult hospitalized patients (66 males, 33 females, median age 57.5 years) received a once-daily injection of 2 g cefodizime for LRTI. Median treatment duration was 8 days. Forty-two patients received cefodizime intravenously and 57 intramuscularly. Indications for treatment were as follows; primary lobar pneumonia (n = 36), bronchopneumonia (n = 14), secondary pneumonia (n = 3), aspiration pneumonia (n = 5), acute exacerbation of chronic bronchitis (n = 21), and of bronchiectasis (n = 9) and acute purulent bronchitis (n = 11). General condition was good in 29 patients and poor in 58; 12 patients were critically ill. The following pathogens were isolated at baseline (source: bronchial secretions, sputum or blood): S. pneumoniae (n = 47), Haemophilus spp. (n = 17), M. catarrhalis (n = 6), Streptococcus spp. (n = 9), Staphylococcus spp. (n = 5), Klebsiella spp. (n = 4), Pseudomonas spp. (n = 1), A. calcoaceticus (n = 1) and anaerobic organisms (n = 7). Fifty-nine patients were evaluable for bacteriological response and 82 for clinical response. Bacteriological outcome was satisfactory in 29/30 patients having LRTI with parenchymal involvement (97%) and in 29/29 patients without parenchymal involvement (100%). Clinical cure was achieved in 41/43 evaluable patients with parenchymal involvement (95%) and in 37/39 patients without parenchymal involvement (95%) in the per-protocol analysis and in 54/58 patients (93%) and 37/41 patients (93%), respectively, in the clinical intention-to-treat analysis. Three of the patients with an unsatisfactory clinical response died of infection during the study. Cefodizime was well tolerated. Adverse reactions--all of mild intensity--were tachycardia, lumbalgia and dizziness, each occurring in one patient. Cefodizime 2 g once daily either i.m. or i.v. was effective in the treatment of lower respiratory tract infections in hospitalized patients. PMID- 9205787 TI - Influence of tetracyclines or cetylpyridinium bromide on activity of fluoroquinolones in Euglena gracilis. AB - To verify the hypothesis that the fluoroquinolone inhibitors of DNA gyrase generate some oxidant species and subsequently free radicals, the effects of simultaneous action of fluoroquinolones (ofloxacin, fleroxacin) and tetracyclines (tetracycline, doxycycline, thiatetracycline) or cetylpyridinium bromide as possible antioxidants on the flagellate Euglena gracilis were examined. Chloroplasts due to their bacterial character were injured and subsequently eliminated by fluoroquinolone. The loss of chloroplasts was accompanied by bleaching of originally green euglena cells. Tetracyclines, as well as cetylpyridinium bromide, decreased the euglena bleaching caused by ofloxacin or fleroxacin. The bleaching induced by ofloxacin or fleroxacin was most effectively inhibited by thiatetracycline and cetylpyridinium bromide. Control treating of cells with tetracyclines or cetylpyridinium bromide alone did not exert any bleaching effect. In vitro weak but significant interactions between examined substances were established as well. PMID- 9205788 TI - Lateral hypothalamic stimulation inhibits dentate granule cell LTP: direct connections. AB - We discovered that angiotensin II (Ang II) applied directly to the dentate gyrus inhibited LTP induction in medial perforant path-dentate granule cell synapses and that the inhibition can be blocked by losartan, an Ang II AT1 receptor specific antagonist. In the first part of this study we found that electrical stimulation of the lateral hypothalamus (LH) inhibits LTP in these synapses and the inhibition can be blocked by pretreating the animals with losartan, indicating that LH angiotensin-containing neurons project to the dentate gyrus. Results of the second part of the study demonstrate clearly that some angiotensin containing LH neurons project directly to dentate granule cells. LH neurons were identified by retrograde tracers applied to the granule cell layer. Double labeled neurons containing angiotensin and HRP were sparsely distributed and both fusiform and multipolar LH neurons appeared in a small cluster lateral and ventral to the fornix at the level of the paraventricular nucleus. Large numbers of angiotensin staining neurons were observed in the hypothalamus. Results support our hypothesis that some angiotensin containing LH neurons project directly to the dentate gyrus. PMID- 9205789 TI - Effects of systemic administration of 2-(4-phenyl-piperidino)-cyclohexanol (vesamicol) and an organophosphate DDVP on the cholinergic system in brain regions of rats. AB - Vesamicol is known to inhibit the transport of acetylcholine (ACh) into synaptic vesicles in vitro, but much less is known about its effects in the brain in vivo. To assess the effect of vesamicol in vivo, we examined cholinergic parameters, such as the subcellular distribution of ACh, activities of enzymes, uptake of choline, and muscarinic receptor binding in the striatum, hippocampus, and cerebral cortex of rats 30 and 60 min after intraperitoneal injection of vesamicol (3 mg/kg) or of vesamicol in combination with DDVP (5 mg/kg), which was administered 10 min before vasamicol. The levels of cytosolic ACh increased in all regions of the brain after injection of vesamicol, while those of vesicular ACh decreased in all regions except for the striatum. The increase in the levels of extracellular ACh and cytosolic ACh in the striatum induced by DDVP was generally enhanced after injection of vesamicol, Vesamicol did not reduce the level of vesicular ACh when DDVP had been injected previously. Vesamicol did not induce any significant changes in the activities of enzymes, choline uptake, or binding of [6H]quinuclidinyl benzilate to the muscarinic ACh receptors in the three regions. Changes in the cholinergic parameters caused by DDVP were not reversed by the combined administration of DDVP with vesamicol. The present results indicate that vesamicol can inhibit the transport of ACh into synaptic vesicles in the brain tissue in vivo, although it cannot reverse the effects of DDVP that has been injected prior to vesamicol. PMID- 9205790 TI - Monoamine neurotransmitter metabolites and spontaneous recurrence of methamphetamine psychosis. AB - The present study was conducted to evaluate plasma levels of monoamine neurotransmitter metabolites in spontaneous recurrences of methamphetamine psychosis (i.e., flashbacks). The subjects were 50 physically healthy females comprised of 25 who experienced flashbacks (flashbackers), 18 who did not experienced methamphetamine psychosis, and 9 who were currently suffering from persistent methamphetamine psychosis. The control data were available from 28 normal healthy females, of whom 20 had previously abused methamphetamine (users) and 8 who had not (nonusers), none of whom had ever become psychotic. Plasma levels of norepinephrine (NE), dopamine (DA), and 5-hydroxytryptamine (5-HT), and their respective metabolites were assayed. Plasma NE levels were significantly higher in the 25 flashbackers during their flashbacks than during their periods of normalcy, and were significantly higher than those in the 20 user and 8 nonuser controls. Plasma 3-methoxy-4-hydroxyphenylglycol (MHPG) levels during flashbacks were significantly higher than those in the 20 user controls. The nine subjects with persistent methamphetamine psychosis had significantly higher NE levels than the user and nonuser controls. The 16 nonflashbackers had significantly higher MHPG levels than the user controls. The present study suggests that an increase in peripheral noradrenergic activity is related to the occurrence of flashbacks. PMID- 9205791 TI - Molecular genetics of hereditary dystonia--mutations in the GTP cyclohydrolase I gene. AB - We found that mutations of GTP cyclohydrolase I, the rate-limiting enzyme in the biosynthesis of tetrahydrobiopterin, which is the cofactor of dopamine synthesizing tyrosine hydroxylase, cause dominantly inherited hereditary progressive dystonia with marked diurnal fluctuation (HPD, Segawa's disease) probably owing to the decrease of dopamine in the basal ganglia. These results indicate that tyrosine hydroxylase in the nigrostriatal dopamine neurons may be most sensitive to tetrahydrobiopterin deficiency causing dystonia. PMID- 9205792 TI - Ameliorating effects of SDZ ENA 713 on age-associated decreases in learning performance and brain choline acetyltransferase activity in rats. AB - In the present study, we have investigated the effects of SDZ ENA 713 on spatial learning deficits in aged rats. Using the same animals, the effect of SDZ ENA 713 on choline acetyltransferase was simultaneously studied to obtain a basis for the behavioral study. In the aged rats, the spatial learning and choline acetyltransferase activity in the frontal cortex were significantly deteriorated compared with young adult rats. SDZ ENA 713 (0.2 mg/kg) significantly shortened the time to reach a hidden platform without affecting swim rates in the water maze task. SDZ ENA 713 (0.1 and 0.2 mg/kg) inhibited aging-induced decreases in choline acetyltransferase activity in the frontal cortex. These results suggest that SDZ ENA 713 ameliorates aging-induced learning deficits and cholinergic dysfunction in rats. PMID- 9205793 TI - Changes in the albumin binding of tryptophan during postoperative recovery: a possible link with central fatigue? AB - Tryptophan is the precursor of the neurotransmitter 5-hydroxytryptamine (5-HT), known to be involved in sleep and fatigue. In the blood, tryptophan binds to albumin, and that which does not, free tryptophan, competes with branched chain amino acids (BCAA) for entry into the brain. The plasma concentrations of albumin, free tryptophan, total tryptophan, and BCAA were measured before and after major surgery in nine elderly and nine coronary artery bypass graft (CABG) patients. In both the elderly and the CABG patients plasma free tryptophan concentrations were increased after surgery, compared with baseline levels; the plasma free tryptophan/BCAA concentration ratio was also increased significantly after surgery. Plasma albumin concentrations were decreased significantly after surgery in both the elderly and the CABG patients. Plasma BCAA concentrations were not affected by surgery in either group. The effect of exercising to exhaustion on 5-HT and tryptophan were investigated in Nagase analbuminemic rats (NAR). The intrasynaptosomal concentration of tryptophan, 5-hydroxy-tryptophan, and 5-HT was increased by fatigue after exercise. In addition, running time to exhaustion was shortened in NAR. These data suggest that free tryptophan uptake and 5-HT synthesis were enhanced in the nerve terminal. A decrease in plasma albumin may account for the increase in plasma-free tryptophan levels. An increase in plasma free tryptophan, resulting in an enhanced plasma concentration ratio of free tryptophan/BCAA, may lead to a higher 5-HT concentration in some parts of the brain and, consequently, to central fatigue. It is suggested that provision of BCAA as a dietary supplement may counteract the increase in plasma free tryptophan and thus improve the status of some patients after major surgery. PMID- 9205794 TI - Acetylcholine: oscillation of levels in mouse brain following electroshock. AB - The acetylcholine contents of mouse brain regions were measured in order to investigate the response of cholinergic neurons to electroshock. In this study, mice were subjected to electroshock and then sacrificed by microwave irradiation at time intervals of from 0.4 to 6.9 a after electroshock. In all of the brain regions studied, the acetylcholine concentration appeared to oscillate with a mean period of approximately 2.5 a following electroshock. The rate of recovery of acetylcholine after electroshock was calculated for each brain region from the oscillatory equation obtained by nonlinear regression analysis of the experimental data. The rates of synthesis of acetylcholine derived from these in vivo measurements were substantially higher than have been indicated by other methods. PMID- 9205795 TI - Microwave fixation and localization of calcium in synaptic terminals using x-ray microanalysis and electron energy loss spectroscopy imaging. AB - The distribution of calcium ions is demonstrated in synaptic terminals by means of a two-step chemical precipitation of calcium ions in the rat brain. K oxalate/K-antimonate chemical replacement with simultaneous computerized microwave irradiation was used. This precipitate in nerve cell structures was investigated by computerized electron probe x-ray microanalysis (EDX) and electron energy loss spectroscopic (EELS) imaging. The values obtained by EDX agreed with those of the standard sample and theoretical values of Ca-antimonate. Typical EELS spectra of Ca:L, O:K, and Sb:M were obtained from nerve terminals in the same tissue block as that used for EDX analysis. Excellent net Ca:L and Sb:M EELS digital images were obtained after their background images were subtracted. Calcium ions were distributed in the nerve terminals, synaptic vesicles, mitochondria, and synaptic membranes. PMID- 9205796 TI - Characterization of alpha, beta-methylene ATP binding sites in mouse crude synaptic membranes. AB - Since ATP has been reported to be a potent excitatory transmitter in the mammalian central nervous system (CNS), we studied the neurochemical characters of the binding sites of alpha,beta-methylene ATP, an agonist of P2x receptors, in mouse crude synaptic membranes. ATP and its related compounds inhibited [3H] alpha,beta-methylene ATP binding in a concentration-dependent manner. The potency order in the inhibition of the binding was as follows; alpha,beta-methylene ATP = ADP beta S > ATP gamma S > ATP > or = ADP > beta,gamma-methylene ATP >> UTP > 2 methylthio ATP. And adenosine did not affect the binding. The order was different from those reported in peripheral tissues. And Sr2+, Ca2+, Mg2+, and Cd2+ enhanced the binding. These results suggest that alpha,beta-methylene ATP binding sites in CNS have different characters from those in peripheral tissues. PMID- 9205797 TI - Hypothalamic cholinergic activity and 2-deoxyglucose-induced hyperglycemia. AB - To clarify the role of the hypothalamic cholinergic system in the regulation of peripheral glucose metabolism, we investigated hypothalamic cholinergic activities after administration of 2-deoxyglucose (2-DG). Intravenous administration of 2-DG (500 mg/kg) caused neuroglycopenia and marked hyperglycemia; the level of plasma glucose increased to 210% of the initial levels of 20 min. For evaluation of the cholinergic activity, we employed a microwave device and subsequently analyzed the contents of acetylcholine (ACh) and choline after microdissection of the hypothalamic nuclei, ventromedial hypothalamic nucleus (VMH), lateral hypothalamus (LH), and paraventricular nucleus (PVN). In addition, we analyzed fluctuation of extracellular levels of ACh using in vivo brain microdialysis. A decrease in the ACh content, and a corresponding increase in the choline content, was observed in those hypothalamic nuclei min after administration of 2-DG. In the microdialysis perfusate, on the other hand, extracellular level of ACh was increased by 2-DG administration. These data show that ACh release, which is cholinergic activity, was increased after 2-DG administration. Our results suggest the involvement and importance of the hypothalamic cholinergic system in 2-DG-induced hyperglycemia. PMID- 9205798 TI - Time-course alterations of monoamine levels and cerebral blood flow in brain regions after subarachnoid hemorrhage in rats. AB - To investigate the possible correlation between changes in monoaminergic neuronal activity and cerebral blood flow (CBF) in the same brain region after subarachnoid hemorrhage (SAH), monoamine levels were analyzed by both HPLC-ECD and fluorohistochemistry techniques, and CBF was measured by using colored microspheres. At the second day of SAH, significant and nonsignificant reductions in blood flow were seen in the examined brain regions with a marked increase in CBF appearing in the telencephalon and hypothalamus on the third day. Significant reductions of monoamine levels in most brain regions were also observed on the second day after SAH, whereas norepinephrine (NE) levels in midbrain increased to 1.5 times compared to the normal level. These reductions were sustained until the fourth day of SAH, although at the third day, serotonin (5-HT) and dopamine levels in the hippocampus and 5-HT levels in the cerebelium were significantly elevated. In fluorohistochemical studies, the fluoro-intensities of monoamines, particularly catecholamines, in the midbrain dorsal NE bundle were enhanced at the second day after SAH. These NE neurons originated from the A5 cell group close to the area where homologous blood was applied through the cisterna magna. The results obtained after SAH show an apparent correlation between changes in monoamine levels and CBF in norepinephrine (NE)-rich areas. These results suggest that SAH-induced neuronal dysfunctions, particularly with NE neurons, are caused not only by reductions of blood flow but also by hemorrhage. PMID- 9205800 TI - Changes in cholinergic neurons and failure in learning function after microsphere embolism-induced cerebral ischemia. AB - Central cholinergic neurons play an important role in learning and memory functions. The present study was undertaken to elucidate the pathological changes in learning function and acetylcholine metabolism of the cerebral cortex and hippocampus, following microsphere embolism in rats. Microspheres (48 microns) were injected into the right internal carotid artery of the rats. Learning function was determined using a passive avoidance task on the seventh day after the embolism. In the biochemical study, acetylcholine and choline contents, and choline acetyltransferase activity were measured in the cerebral cortex and hippocampus. Cortical acetylcholinesterase-containing fibers were quantitatively estimated in the embolized rat. Passive avoidance was impaired in the microsphere embolized rat. Microsphere embolism decreased the acetylcholine concentration and choline acetyltransferase activity in the cerebral cortex and hippocampus. In the histochemical study, the length of cortical acetylcholinesterase-containing fibers was decreased, but cell density was unchanged in the ipsilateral hemisphere of the microsphere-embolized rat. The results suggest that microsphere embolism induces severe damage to cholinergic neurons, which may be related to the impairment of learning function in the ischemic brain. PMID- 9205799 TI - Effects of MK-801 and pentobarbital on cholinergic terminal damage and delayed neuronal death in the ischemic gerbil hippocampus. AB - The present study covers both the effects of MK-801, a noncompetitive N-methyl-D aspartate (NMDA) receptor antagonist, and pentobarbital on cholinergic terminal damage and delayed neuronal death (DND) in ischemic gerbil. To study the above effects, in vivo microdialysis, immunohistochemical, and morphological techniques were used. MK-801 (3 mg/kg) or pentobarbital (50 mg/kg) were injected intraperitoneally 1 h or 30 min before 5 min ischemia, respectively. Each estimation was then carried out 4, 7, or 14 days after ischemia. Ischemia induced a significant decrease in acetylcholine (ACh) release and a disappearance of choline acetyltransferase (ChAT)-immunoreactivity in the hippocampus in addition to inducing DND. On day 4, MK-801 protected ischemia-induced DND in the hippocampal CA1 subfield. However, MK-801 had no effect against the decrease in ACh release in spite of protection of the decrease in ChAT-immunoreactivity. On day 7 and 14, no protective effect of MK-801 was observed in all estimations. It became clear that the mechanism of cholinergic terminal dysfunction is different from that involved in pyramidal cell death, i.e., excitative neurotoxicity induced by overabundant extracellular glutamate. Pentobarbital also provided protection against DND. However, protective effects of pentobarbital on the decrease in ACh release and the low ChAT-immunoreactivity were incomplete. Our present study indicated a limitation on the efficacy of NMDA receptor antagonist and barbiturate against cerebral ischemia. PMID- 9205801 TI - Selective full dopamine D1-like (SKF-82958) and D2-like (N-0923) agonist combination in the MPTP monkey model of hemiparkinsonism. AB - Both SKF-82958 and N-0923, selective full D1-like and D2-like agonists, respectively, given IM produced contraversive circling and reduced neurologic deficits in six MPTP-induced hemiparkinsonian monkeys. A small fixed dose of N 0923 (10 micrograms/kg) and increasing doses of SKF-82958 (23.4-234 micrograms/kg) in combination were synergistic or antagonistic in this animal model. A small dose (23.4 micrograms/kg) of SKF-82958, in combination with N 0923, caused potentiation, an intermediate dose (74.8 micrograms/kg) in combination produced additive effects, while a very large dose (234 micrograms/kg) in combination produced antagonism. All three doses of SKF-82958 prolonged the duration of action of a small dose (10 ng/kg) of N-0923. Selective D1-like and D2-like agonists should be studied as potential therapeutic agents alone and in combination in human idiopathic parkinsonism, especially using low and intermediate doses. PMID- 9205802 TI - Effects of postmethamphetamine treatment with restraint on ambulatory sensitization to methamphetamine in mice. AB - The administration of methamphetamine (2 mg/kg SC) to mice was followed by acceleration of ambulation (locomotion) for 3 h, with the peak effect at 1/2-1 h. When mice were allowed free ambulation for 3 h in an activity cage of 20 cm in diameter after methamphetamine administration, repeated administrations of methamphetamine, four times at 3-day intervals, caused ambulatory sensitization; the count at the fourth administration being approximately 2.3 times as high as that at the first administration. Furthermore, the mice that were allowed ambulation in the activity cage during postmethamphetamine period of either 0 1/2, 0-1, 0-2, or 1/2-1 h (during the other periods, the mice were put in jars of 6 cm in diameter to restrict ambulation) demonstrated ambulatory sensitization to methamphetamine, and these levels were as same as that in the mice given methamphetamine with free ambulation for 3 h. However, the free ambulation during postmethamphetamine periods of 0-1/4 h and 1-3 h failed to produce strong sensitization to methamphetamine. These results suggest that a free ambulation for at least 1/2 h during the postmethamphetamine period of 0-1 h is important for induction of ambulatory sensitization to methamphetamine in mice. PMID- 9205803 TI - Alterations in the spontaneous release of dopamine and the density of the DA D2 receptor mRNA after chronic postnatal exposure to cocaine. AB - The influence of cocaine administration on dopamine (DA) release and D2 dopamine receptor mRNA levels was examined in developing rat brain. In the rat pup, cocaine (25 mg/ kg SC) was administered daily from postnatal days 1-9 and extracellular DA measured 24 h after the last injection of cocaine, using in vivo micro dialysis. Twenty-four hours after discontinuing cocaine administration, a decrease in the extracellular concentration of DA of more than 100% was found in treated pups compared to control pups. Pups were tested on postnatal days 10-12, 20-21, or 35-36. After 1 month, basal release of DA returned to control levels. To examine the structural basis of the alteration in basal release of DA, in situ hybridization studies were performed to access the effect of chronic administration of cocaine on the mRNA encoding the D2 DA receptor. These preliminary studies, on postnatal day 10, indicate that drug treatment alters the developmental pattern of D2 mRNA. The changes in D2 mRNA expression were accompanied by delayed disaggregation of neostriatal cells and diminished growth of neostriatal neurons. These structural changes may lead to functional impairment in the development of dopamine target cells, thus altering the balance of synaptic and trophic effects of DA. PMID- 9205804 TI - Regulation of dopamine D1 and D2 receptors on striatal acetylcholine release in rats. AB - The effects of dopamine (DA) D1 and D2 receptors on striatal acetylcholine (ACh) releases were investigated by in vivo microdialysis. All drugs were applied via dialysis membrane directly to the striatum. The levels of ACh release were increased by 10(-4) M SKF38393, a D1 receptor agonist. Although 10(-4) M SCH23390, a D1 receptor antagonist, exhibited an increase in the levels of ACh release, the agonist (10(-4) M) induced-increase in the levels of ACh release was suppressed by coperfusion of the antagonist (10(-4) M). In contrast, the levels of ACh release were decreased by the D2 receptor agonist, N-434, in a dose dependent manner (10(-4) M to 10(-7) M) and increased by the D2 receptor antagonist, sulpiride, in a dose-dependent manner (10(-5) M to 10(-7) M). The agonist (10(-5) M) induced-decrease in the levels of ACh release was suppressed by coperfusion of the antagonist (10(-4) M). Coperfusion of D1 (10(-4) M) and D2 (10(-5) M) agonists blocked both effects of respective drug alone. In order to clarify the effect of endogenous DA, two drugs with different mechanisms for enhancing DA concentration in the synaptic cleft, the DA release-inducer methamphetamine, and the DA uptake inhibitor nomifensine were perfused separately. Both (10(-4) M to 10(-5) M) produced a dose- and a time-dependent decrease in the levels of ACh release. Significant higher levels of ACh release were observed in the striatum of the 6-hydroxydopamine (8 micrograms/10 microliters)-treated rats with significant depletion of striatal DA content. These results suggest that in striatal DA-ACh interaction ACh release, as cholinergic interneuron's activity, is tonically inhibited via the D2 receptor, mainly by dopaminergic input, and the D1 receptor probably modifies the effect of the D2 receptor indirectly. PMID- 9205805 TI - Blockade of hippocampal long-term potentiation following soman pretreatment in the rat. AB - One of the most potent toxins known is the cholinesterase inhibitor, soman, which produces severe convulsions and cell loss in the central nervous system. In these experiments the effect of multiple low doses of soman on the acquisition and maintenance of long-term potentiation (LTP) was determined in rats. LTP is a form of synaptic plasticity that has been studied as a cellular substrate for learning and memory mechanisms. Under urethane anesthesia, electrodes were positioned in the dentate gyrus for recording evoked potentials before and after tetanic stimulation of the perforant path. LTP levels were greatly reduced in rats recovering from convulsion-inducing soman treatment. Rats exposed to similar amounts of soman, but not displaying convulsions, also displayed reduced levels of LTP. The responses recorded from these animals were highly variable, ranging from control levels of potentiation to no LTP. The variability could be attributed to some animals having convulsions that were not detected before surgery or to other interanimal differences in the degree of soman-induced toxicity. The long range goal of the experiments presented here is to develop a better rodent model for studying soman-induced functional changes in the CNS that can be detected prior to the gross morphological changes. PMID- 9205806 TI - Influence of age on the cerebral lesions in an immature rat model of cerebral hypoxia-ischemia: a light microscopic study. AB - The most frequently used model of neonatal cerebral hypoxia-ischemia consists of a 7-day postnatal rat model with combined common carotid artery ligation and hypoxemia. Neuropathologic studies have shown major differences between this 7 day postnatal rat model and a similar adult model in regard to overall cerebral vulnerability, type and distribution of lesions. It is not clear how and when during animals' development these changes in cerebral vulnerability take place. To determine this we studied groups of rats of 2 to 30 postnatal days. The animals underwent unilateral common carotid artery ligation followed by breathing in 8% oxygen for 30, 60, 90, or 120 min and their brains were examined at 24- or 72-h recovery intervals. Due to resistance of 2-3-day-old rats to develop cerebral hypoxic-ischemic damage, 5% O2 was used instead of 8% O2. The results indicate that: (i) There is an overall increase in severity of cerebral lesions on the side of common carotid artery ligation between 2 and 7 postnatal days. There is also an increase in the frequency of cerebral lesions in developing animals with increasing age. (ii) Hippocampus is remarkably resistant to hypoxic ischemic insult at 2-3 postnatal days but becomes progressively vulnerable, and by age 13 postnatal days hippocampal vulnerability far exceeds that of cortex. (iii) Cortical lesions change from predominantly columnar cell death to laminar selective neuronal death at age 13 postnatal days. (iv) Also significant changes occur in relative vulnerability of various hippocampal regions during development. During the first 5 postnatal days relative vulnerability of hippocampal regions is similar, but as the animals' development proceeds and hippocampal vulnerability increases lesions tend to involve specific regions while sparing others. By age 13 postnatal days CA1 and lateral CA3 develop increased vulnerability while medial CA3 and fascia dentata become relatively resistant and by 21 postnatal days adult pattern of CA1 selective vulnerability is approached. The underlying mechanisms for these changes in regional vulnerability to cerebral hypoxia-ischemia during development should be sought in complex regional anatomic, functional, and metabolic alterations that take place as brain matures. PMID- 9205807 TI - Ontogenic development of secretogranin II and of its processing to secretoneurin in rat brain. AB - The ontogenic development of secretogranin II was studied by immunochemistry and immunohistochemistry. Extracts of brains from various developmental stages were analyzed by a radioimmunoassay for secretoneurin, a peptide derived from secretogranin II. From gestational day 13 to adulthood the levels increased from 0.1 to 94 fmol/mg wet weight. Characterization of the immunoreactivity by molecular sieve chromatography revealed that throughout all developmental stages the proprotein secretogranin II was fully processed to the free peptide secretoneurin. In immunohistochemistry secretoneurin-IR was first detected at embryonic day 13. Between embryonic days 14 and 18 a strong increase in the number of secretoneurin immunopositive cells was observed in many brain areas, notably in the amygdala, hypothalamus, olfactory bulb and several brainstem nuclei. The pattern of staining during development is quite similar to that in the adult. The present paper demonstrates that secretoneurin immunoreactivity appears early in embryonic life. Processing of the proprotein secretogranin II starts when the protein is first synthesized apparently at about the same time when the prohormone convertase PC1 and PC2 can be demonstrated. PMID- 9205808 TI - Identification of a chicken homologue in the Brn-3 subfamily of POU-transcription factors. AB - Among the many transcription factors thus far identified several are found to be expressed almost exclusively in the nervous system. The Brn-3 subfamily of POU transcription factors constitutes a highly conserved group of such factors showing expression predominantly in sensory neurons. We now describe the nucleotide sequence and proposed amino acid sequence of a chicken homologue to the murine and human Brn-3 genes. Furthermore we characterise the early embryonic expression pattern of this chicken Brn-3 gene and show it to be expressed in peripheral sensory ganglia as well as in retinal ganglion cells. Based on these findings we conclude that the chicken homologue to the murine and human Brn-3a genes has been cloned. We have begun to examine possible regulatory pathways of Brn-3a by stimulating chick embryonic peripheral ganglia with trophic factors and assaying resulting levels of Brn-3a with a quantitative PCR approach. Trigeminal and dorsal root ganglia stimulated in culture by NGF and NT-3 embryonic day 9 (E9) produce neurites without raising the Brn-3a mRNA levels. PMID- 9205809 TI - Growth cone interactions with purified cell and substrate adhesion molecules visualized by interference reflection microscopy. AB - The migration of growth cones on substrates consisting of naturally occurring cell adhesion molecules has been extensively studied in cell culture. However, relatively little is known about how growth cones contact the substrate or how the patterns of contact change as growth cones move forward. We have examined the interactions of chick retinal ganglion cell growth cones with laminin, merosin, N cadherin, L1 and poly-L-lysine by time-lapse interference reflection microscopy (IRM) using a laser scanning confocal microscope. In images obtained by IRM, areas of a cell that are closely apposed to the substrate appear dark whereas areas that are farther away appear light. Growth cones on Jaminin and merosin were almost uniformly light, indicating that very little of the membrane was in close contact with the substrate. Growth cones on N-cadherin had a mottled appearance with some relatively large dark gray areas. The proximal portions of filopodia often were dark, in contrast to those on laminin and merosin which were light. In addition, growth cones on N-cadherin had numerous dark gray punctate regions of close association with the substrate. Growth cones on L1 had darker regions than growth cones on other substrates and these comprised a larger fraction of their area. There also were differences in the temporal dynamics of growth cone interactions with different substrates and these differences correlated with differences in rates of growth. None of the contacts observed in growth cones were as dark or stable as focal contacts of fibroblasts. PMID- 9205810 TI - Sexually dimorphic effects of maternal adrenalectomy on hypothalamic corticotrophin-releasing factor, glucocorticoid receptor and anterior pituitary POMC mRNA levels in rat neonates. AB - The stress hyporesponsive period (SHRP) occurs during the first two weeks of life (from day 4 to day 14) in the rat. SHRP may occur due to immature hypothalamic pituitary-adrenal (HPA) regulatory mechanisms of the neonate. Decreased expression of corticotropin-releasing factor (CRF) has been observed during this period, and this decreased hypothalamic 'drive' may contribute to the manifestation of SHRP in the rat neonate. Since maternal corticosteroids are elevated toward the end of gestation they may suppress fetal CRF expression leading to a less than adequate activation of the neonatal hypothalamus in response to stress. Therefore, the purpose of the present study was to clarify the contribution of maternal glucocorticoids to the decreased CRF expression observed during SHRP in the neonatal rat. We investigated the effects of maternal adrenalectomy on hypothalamic CRF, glucocorticoid receptor (GR) and anterior pituitary proopiomelanocortin (POMC) mRNA levels in male and female neonates. Maternal adrenalectomy, or sham surgery, was performed on day 8 of gestation and the mRNA levels of CRF, GR and POMC were measured by Northern blotting in the offspring on their postnatal days 1, 7, 14, and 21. The observed changes in the mRNA levels of these genes suggests that an important developmental event occurs in the regulation of these HPA genes between neonatal days 7 and 14 corresponding to the termination of SHRP. Female offspring had significantly higher levels of CRF mRNA than males throughout this period. The lack of maternal corticosteroids evoked a gender-specific response in the neonates. In female offspring, maternal adrenalectomy resulted in a dramatic and correlated increase in mRNA levels of hypothalamic CRF and GR on day 14, with pituitary POMC expression not following this increase. There was no significant effect of maternal adrenalectomy on the expression of these genes in males. These results suggest a sex difference in response to maternal glucocorticoids in the fetus. However, the role of maternal corticosteroids in the low expression of CRF during SHRP could not be established from this study, since their removal by adrenalectomy did not advance the expression profile of CRF toward an earlier increase in the neonatal hypothalamus. PMID- 9205811 TI - Maturation of the blood-retina barrier in the developing pecten oculi of the chicken. AB - The major interest in the development of the blood-brain barrier and its underlying induction mechanisms is given by the crucial role they play in the maturation of the central nervous system in general. Whilst it is believed that it is the microenvironment in the brain that destines the endothelial cells to become committed to barrier properties, the analysis of the multitude of factors probably responsible for this commitment is extremely difficult. Therefore, in a previous study, we inaugurated the pecten oculi of the avian eye as a relatively simple in vivo model of the blood-brain barrier [Gerhardt, S. et al, Cell Tissue Res., 285 (1996) 91-100]. In the present study, we demonstrate data on the development of the pecten which allow us to understand better the commitment of barrier properties in endothelial cells in an environment which is considerably less complex than that realized in the brain. The pecten is built up by mainly two cell types, the pigmented glial cells and the endothelial cells. The pigmented cells, which are believed to originate from the retinal pigment epithelium, lose their tight junctions in the microenvironment of the vitreous body, whereas the endothelial cells, which originate from the permeable choroidal vessels, gain tight junctions and other barrier properties in the microenvironment of the vitreous body. On embryonic day 7 (E7), tight and gap junctions between epithelial-like glial cells line the vitreal border of the developing pecten. By E16, these junctions disappear, and the endothelial cells gradually acquire barrier characteristics (continuous and P-face associated tight junctions, no extravasation of lanthanum nitrate, and the exclusive expression of the glucose transporter isoform 1 and the barrier specific antigen HT7 in their luminal and abluminal membranes). The results are discussed considering the switch from an epithelial (glial) to an endothelial barrier. PMID- 9205812 TI - Cocaethylene exposure during the brain growth spurt period: brain growth restrictions and neurochemistry studies. AB - The concurrent use of alcohol and cocaine has recently attracted attention in the medical research field due to the prevalence of this drug abuse pattern and the exclusive formation of a pharmacologically active substance, cocaethylene (CE). This is the first study to examine the neuroteratogenic effects of cocaethylene exposure during the brain growth spurt (part of the third trimester equivalent) on brain growth restrictions and neurochemical profiles. For the brain growth restrictions study, three groups of artificially reared rat pups were given daily injections of 0, 10 or 20 mg/kg cocaethylene (s.c.) from postnatal days (PDs) 4 through 9. One group of normally reared pups (suckle control) also was used. These pups were perfused on PD 10 and the brains were removed and weighed (forebrain, cerebellum and brainstem). For the neurochemistry study, five groups of artificially reared pups were used and were treated identically to those in the brain growth restrictions study, with the exceptions that animals assigned to acute cocaethylene treatment groups did not receive cocaethylene from PDs 4 through 8 and all animals in this study were sacrificed on PD 9 by decapitation. One suckle control group was included to control the possible artificial rearing effects on the neurochemical measures. Blood and fresh brain tissues (cortex, subcortical structures, cerebellum and brainstem) were collected for blood cocaethylene concentration and neurochemical analyses using GC/MS and HPLC techniques, respectively. The statistical analyses indicated that daily administration of 10 or 20 mg/kg cocaethylene, but not 0 mg/kg cocaethylene, significantly restricted the brain growth (brain weights) in all three brain regions assessed. Furthermore, cocaethylene administration from PDs 4 through 9 produced region-specific alterations in various neurotransmitter concentrations. The changes in neurotransmitter levels were not a function of the responses to the last cocaethylene injection on PD 9, since the outcomes between six days of cocaethylene treatment (PDs 4 to 9) and one day acute treatment (PD 9) were notably different. Furthermore, the artificial rearing procedure appeared to produce significant alterations in various neurotransmitter levels when compared with normally reared (suckle) controls. Collectively, these results suggest that cocaethylene is neuroteratogenic to the developing brain during the third trimester equivalent and the unique formation of cocaethylene resulting from the concurrent use of alcohol and cocaine may represent an increased risk to the developing brain beyond the intrinsic neuroteratogenic effects of cocaine and alcohol individually. PMID- 9205813 TI - Sodium-dependent proline and glutamate uptake by hippocampal synaptosomes during postnatal development. AB - NA(+)-dependent uptake of proline and glutamate by hippocampal synaptosomes was studied during postnatal development. At all ages from 9 days to adulthood, hippocampal synaptosomes transported proline by both a high-affinity and a low affinity process, whereas glutamate was always transported predominantly by a high-affinity process. During the period of rapid synaptogenesis, the KT for high affinity proline transport overshot the adult value, whereas the KT for glutamate transport increased steadily toward the adult value. The ratio of KT values for proline and glutamate was 2-3 times the adult value between 12 and 24 days of age. Although high-affinity transporters for proline and glutamate are expressed by nearly the same hippocampal pathways, they are differentially regulated during postnatal development. PMID- 9205814 TI - Pharmacoeconomics: integrating economic evaluation into clinical trials. PMID- 9205815 TI - Pharmacokinetics of beta-adrenoceptor blockers in obese and normal volunteers. AB - AIMS: Obesity can modify the pharmacokinetics of lipophilic drugs. As beta adrenoceptor blockers (BB) are often prescribed for obese patients suffering from hypertension or coronary heart disease, this study compares the pharmacokinetics of lipophilic beta-adrenoceptor blockers in obese and control subjects. METHODS: Nine obese (157 +/- 24% of ideal body weight (IBW) mean +/- s.d.) and nine non obese healthy volunteers (98 +/- 10% IBW), aged 32 +/- 9 years, were included in the study. Subjects were randomly given a single i.v. infusion of one of the following racemic beta-adrenoceptor blockers, whose doses (expressed as base per kg of IBW) were: propranolol (0.108 mg), labetalol (0.99 mg) and nebivolol (0.073 mg). The plasma concentrations of unchanged drugs were measured by h.p.l.c. The ionisation constants and lipophilicity parameters of beta-adrenoceptor blockers were assessed. RESULTS: The pharmacokinetic data for the three drugs were qualitatively similar. There was a trend towards a greater total distribution volume (Vss) in obese patients than in controls. However, Vss expressed per kg body weight was slightly smaller in obese patients. The relationship between Vss and lipophilicity of five beta-adrenoceptor was studied by combining the current results with those previously obtained with a moderately lipophilic drug (bisoprolol) and a hydrophilic one (sotalol). The Vss of the five drugs was positively and well-correlated (r2 = 0.90; P < 0.01) with their distribution coefficient at pH 7.4 (log D7.4), but not with their partition coefficients. The linear regression coefficients for lean and obese subjects were very similar. CONCLUSIONS: Lipophilic beta-adrenoceptor blockers seem to diffuse less into adipose than into lean tissues. All electrical forms of the drugs (i.e. cations, neutral forms, or zwitterions) present at physiological pH contribute to their tissue distribution, in both obese and lean subjects. Their tissue distribution in obese patients could be restricted by the sum of hydrophobic forces and hydrogen bonds they elicit with macromolecules in lean tissues. PMID- 9205816 TI - Pharmacokinetics of recombinant human interleukin-11 (rhIL-11) in healthy male subjects. AB - AIMS: To study the pharmacokinetics of recombinant human interleukin-11 (rhIL-11) in healthy male volunteers following subcutaneous (s.c.) and intravenous (i.v.) administration. METHODS: RhIL-11 was infused intravenously at 10-50 micrograms kg 1 for 1 or 3 h, or administered subcutaneously at 3-50 micrograms kg-1 to volunteers. RhIL-11 was also administered at 3 micrograms kg-1 s.c. once daily for 7 days. Plasma and urinary concentrations were measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: RhIL-11 showed linear pharmacokinetics after both intravenous infusion and s.c. administration. Comparison of t1/2 and MRT values after i.v. administration with those after s.c. administration indicated that rhIL-11 pharmacokinetics after s.c. administration were absorption rate-limited. Bioavailability after s.c. administration was about 65%. Since RhIL 11 was not detected in urine after a single 50 micrograms kg-1 s.c. dose, rhIL-11 was considered to be eliminated by metabolism. There was no significant change in the pharmacokinetic profile of rhIL-11 following repeated s.c. administration. CONCLUSIONS: RhIL-11 demonstrated linear pharmacokinetics at these dose ranges after single and repeated s.c. administration or constant-rate i.v. infusion in healthy volunteers. PMID- 9205818 TI - A comparative population pharmacokinetic analysis for methylprednisolone following multiple dosing of two prodrugs in patients with acute asthma. AB - AIMS: To conduct a randomized, parallel group comparison of the population pharmacokinetics of the two methylprednisolone (MP) prodrugs Promedrol (MP suleptanate) and Solu-Medrol (MP succinate) in patients hospitalized with acute asthma. METHODS: Ninety volunteers were included in the pharmacokinetic analysis. Each volunteer received a dosage regimen of 40 mg (MP equivalents) i.v. 6 hourly for 48 h. The bio-conversion and disposition of a 40 mg (MP equivalent) i.v. dose of either MP suleptanate or MP succinate to MP was modelled as a first order input, and a mono-exponential elimination phase. RESULTS: Population modelling indicated that the only difference in MP pharmacokinetics between MP suleptanate and MP succinate was in the input rate constant (66.0 h-1 vs 5.5 h-1 respectively). Based on individual Bayesian estimates, the exposure of patients to MP was marginally lower for MP suleptanate although the parameter estimates were not significantly different for half-life (2.7 h vs 3.0 h), steady-state AUC (2007.0 ng ml-1 h vs 2321.0 ng ml-1 h) and steady-state Cmax (698.4 ng ml-1 vs 647.8 ng ml-1) for MP suleptanate and MP succinate respectively. CONCLUSIONS: It was concluded that for the multiple dosage regimen used in patients with acute asthma the systemic exposure to MP following dosing with MP suleptanate is similar to that arising from MP succinate. In addition the differences in the pharmacokinetics for the prodrugs resulted in only a small difference in the relative bioavailability of MP for MP suleptanate (0.94) compared with MP succinate. PMID- 9205817 TI - The absolute bioavailability and metabolic disposition of the novel antimigraine compound zolmitriptan (311C90). AB - AIMS: Two open studies in healthy volunteers were conducted to determine the absolute bioavailability and metabolic disposition of zolmitriptan (311C90), a novel 5HT1D agonist for the acute treatment of migraine. METHODS: After an initial test i.v. infusion, bioavailability was assessed by comparison of AUC after an i.v. infusion (3.5 mg) and an oral tablet (10 mg), in six men and six women using a randomised, crossover design. Disposition was studied by administration of a 25 mg capsule, labelled with 100 microCi [14C]-zolmitriptan, to five men and one woman on a single occasion. RESULTS: Zolmitriptan was well tolerated by both i.v. and oral routes. Adverse events were mostly mild, consistent with earlier studies and characteristic of this class of drug. Reports were similar in nature and number after both oral and i.v. dosing. Mean +/- s.d. oral bioavailability was 0.49 +/- 0.24 (0.38 +/- 0.16 in men and 0.60 +/- 0.28 in women). After oral dosing, Cmax and AUC values in women were approximately double those in men. Relative to zolmitriptan concentrations, metabolite concentrations were higher after oral dosing than after i.v., and higher in men compared with women. Half-life was significantly longer after oral dosing (mean 22%, 95% CI 6 35%). Mean +/- s.d. values for CL, V2 and t1/2,z after i.v. dosing (all subjects) were 8.7 +/- 1.7 ml min-1 kg-1, 122 +/- 321 and 2.30 +/- 0.59 h respectively. Following administration of 25 mg [14C]-zolmitriptan, 91.5% of the dose was recovered in 7 days, 64.4 +/- 6.5% in urine and 27.1 +/- 6.0% in faeces. Less than 10% was recovered unchanged in urine, with 31.1 +/- 6.4% recovered as the inactive indole acetic acid metabolite. Most of the faecal material was unchanged zolmitriptan, representing unabsorbed drug. Plasma concentrations of [14C] were slightly higher than those of the summed concentrations of known analytes zolmitriptan, the active N-desmethyl metabolite (183C91), the inactive N-oxide (1652W92) and indole acetic acid (2161W92) metabolites, which accounted for 86% of total plasma radioactivity. No other significant metabolites were detected in plasma. Some minor additional metabolites were detected in urine, none of which contributed more than 5% of the dose. CONCLUSIONS: The data suggest that zolmitriptan undergoes first-pass metabolism and this is more extensive in men than in women. Zolmitriptan has suitable bioavailability for an acute oral migraine treatment and there are no significant unidentified metabolites in man. PMID- 9205819 TI - Pharmacokinetic and pharmacodynamic assessment of bioavailability for two prodrugs of methylprednisolone. AB - AIMS: The aim of this study was to establish whether pharmacokinetic differences between two pro-drugs of methylprednisolone (MP) are likely to be of clinical significance. METHODS: This study was a single-blind, randomized, crossover design comparing the bioequivalence of MP released from the pro-drugs Promedrol (MP suleptanate) and Solu-Medrol (MP succinate) after a single 250 mg (MP equivalent) intramuscular injection to 20 healthy male volunteers. Bioequivalence was assessed by conventional pharmacokinetic analysis, by measuring pharmacodynamic responses plus a novel approach using pharmacokinetic/pharmacodynamic modeling. The main measure of pharmacodynamic response was whole blood histamine (WBH), a measure of basophil numbers. RESULTS: The MP Cmax was less for MP suleptanate due to a longer absorption halflife of the prodrug from the intramuscular injection site. The bioavailability of MP was equivalent when based on AUC with a MP suleptanate median 108% of the MP succinate value (90% CI: 102-114%). For Cmax the MP suleptanate median was 81% of the MP succinate value (90% CI: 75-88%). The tmax for MP from MP suleptanate was delayed relative to MP succinate. The median difference was 200% (90% non parametric CI: 141-283%). The area under the WBH effect-time curve (AUEC) and the maximum response (Emax) were found to be equivalent (90% CI: 98-113% and 93-109% respectively). The maximum changes in other white blood cell counts, blood glucose concentration and the parameters of the pharmacodynamic sigmoid Emax model (EC50, Emax and gamma) were also not significantly different between prodrugs. CONCLUSIONS: MP suleptanate is an acceptable pharmaceutical alternative to MP succinate. The use of both pharmacokinetic and pharmacodynamic response data together gives greater confidence in the conclusions compared with those based only on conventional pharmacokinetic bioequivalence analysis. PMID- 9205820 TI - Population analysis of the non linear red blood cell partitioning and the concentration-effect relationship of draflazine following various infusion rates. AB - AIMS: To investigate the impact of the specific red blood cell binding on the pharmacokinetics and pharmacodynamics of the nucleoside transport inhibitor draflazine after i.v. administration at various infusion rates. It was also aimed to relate the red blood cell (RBC) occupancy of draflazine to the ex vivo measured adenosine breakdown inhibition (ABI). METHODS: Draflazine was administered to healthy volunteers as a 15-min i.v. infusion of 0.25, 0.5, 1, 1.5 and 2.5 mg immediately followed by an infusion of the same dose over 1 h. Plasma and whole blood concentrations were measured up to 120 h post dose, and were related to the ex vivo measured ABI, serving as a pharmacodynamic endpoint. The capacity-limited specific binding of draflazine to the nucleoside transporter located on the erythrocytes was evaluated by a population approach. RESULTS: The estimate of the population parameter typical value (%CV) of the binding constant Kd and the maximal specific binding capacity (Bmax) was 0.385 (3.5) ng ml-1 plasma and 158 (2.1) ng ml-1 RBC, respectively. The non-specific binding was low. The specific binding to the erythrocytes was a source of non-linearity in the pharmacokinetics of draflazine. The total plasma clearance of draflazine slightly decreased with increasing doses, whereas the total clearance in whole blood increased with increasing doses. The sigmoidal Emax equation was used to relate the plasma and whole blood concentration of draflazine to the ex vivo determined ABI. In plasma, typical values (%CV) of Emax, IC50 and Hill factor were 81.4 (1.9)%, 3.76 (9.3) ng ml-1 and 1.06 (3.4), respectively. The relationship in whole blood was much steeper with population parameter typical values (%CV) of Emax, IC50 and Hill factor of 88.2 (2.0)%, 65.7 (2.8) ng ml-1 and 4.47 (5.5), respectively. The RBC occupancy of draflazine did not coincide with the ex vivo measured ABI. The observed relationship between RBC occupancy and ABI was not directly proportional but similar for all studied infusion schemes. CONCLUSIONS: The findings of this study show that the occupancy of the nucleoside transporter by draflazine should be at least 90% in order to inhibit substantially adenosine breakdown in vivo. On the basis of these findings it is suggested that a 15 min infusion of 1 mg draflazine followed by an infusion of 1 mg h-1 could be appropriate in patients undergoing a coronary artery bypass grafting. PMID- 9205821 TI - Pharmacokinetics and safety of JTP-4819, a novel specific orally active prolyl endopeptidase inhibitor, in healthy male volunteers. AB - AIMS: To investigate the pharmacokinetics and safety profile of JTP-4819, (-) (2S)-1-benzylaminocarbonyl-[(2S)-2-glycoloylpyrrolidinyl ]-2 pyrrolidinecarboxamide, a novel specific orally active prolyl endopeptidase (PEP) inhibitor. METHODS: JTP-4819 was given orally to 28 healthy male volunteers at single doses of 30 mg (n = 6), 60 mg (n = 6), 120 mg (n = 6) and placebo (n = 3) and multiple doses of 60 mg three times daily (n = 5) and placebo (n = 2) for 7 days to investigate its safety and pharmacokinetics following a preliminary safety evaluation of 3, 10 and 30 mg doses in six healthy volunteers. With the single dose of 60 mg, a cross-over study was conducted to examine the effect of food on the bioavailability of the drug. The concentrations of JTP-4819 in plasma and urine were determined by electrospray ionization-liquid chromatography/mass spectrometry (ESI-LC/MS) method. RESULTS: In the multiple-dose study, the cholinesterase activity was gradually increased and reached above the normal range on days 4 to 8 in all five subjects given JTP-4819 and gradually returned to normal range after completion of dosing. The elevation of plasma cholinesterase activity was considered to be an action of JTP-4819, but this remains to be verified. There were no other abnormal findings in objective symptoms and laboratory findings including blood pressure, heart rate, electrocardiogram, body temperature, haematology, blood chemistry and urinalysis. The Cmax of JTP-4819 at 30, 60 and 120 mg in fasting state were 474, 887 and 1,649 ng ml-1, respectively, at 1 h after administration, and the t1/2 was about 2 h. AUC increased in proportion to the given doses. The cumulative urinary recoveries within 24 h were approximately 66%, Cmax, AUC, t1/2 and urinary recovery were not affected by food intake. In the multiple-dose study, there was no drug accumulation trend in plasma. CONCLUSIONS: These results indicate that JTP-4819 has acceptable pharmacodynamic and pharmacokinetics profiles for clinical use without any serious adverse events as we verified in healthy young male volunteers. PMID- 9205822 TI - Venlafaxine: in vitro inhibition of CYP2D6 dependent imipramine and desipramine metabolism; comparative studies with selected SSRIs, and effects on human hepatic CYP3A4, CYP2C9 and CYP1A2. AB - AIMS: In order to anticipate drug-interactions of potential clinical significance the ability of the novel antidepressant, venlafaxine, to inhibit CYP2D6 dependent imipramine and desipramine 2-hydroxylation was investigated in human liver microsomes. The data obtained were compared with the selective serotonin re uptake inhibitors, fluoxetine, sertraline, fluvoxamine and paroxetine. Venlafaxine's potential to inhibit several other major P450 s was also studied (CYP3A4, CYP2D6, CYP1A2). METHODS: Ki values for venlafaxine, paroxetine, fluoxetine, fluvoxamine and sertraline as inhibitors of imipramine and desipramine 2-hydroxylation were determined from Dixon plots of control and inhibited rate data in human hepatic microsomal incubations. The inhibitory effect of imipramine and desipramine on liver microsomal CYP2D6 dependent venlafaxine O-demethylation was determined similarly. Venlafaxine's IC50 values for CYP3A4, CYP1A2 CYP2C9 were determined based on inhibition of probe substrate activities (testosterone 6 beta-hydroxylation, ethoxyresorufin O-dealkylase and tolbutamide 4-hydroxylation, respectively). RESULTS: Fluoxetine, paroxetine, and fluvoxamine were potent inhibitors of imipramine 2-hydroxylase activity (Ki values of 1.6 +/- 0.8, 3.2 +/- 0.8 and 8.0 +/- 4.3 microM, respectively; mean +/- s.d., n = 3), while sertraline was less inhibitory (Ki of 24.7 +/- 8.9 microM). Fluoxetine also markedly inhibited desipramine 2-hydroxylation with a Ki of 1.3 +/- 0.5 microM. Venlafaxine was less potent an inhibitor of imipramine 2 hydroxylation (Ki of 41.0 +/- 9.5 microM) than the SSRIs that were studied. Imipramine and desipramine gave marked inhibition of CYP2D6 dependent venlafaxine O-demethylase activity (Ki values of 3.9 +/- 1.7 and 1.7 +/- 0.9 microM, respectively). Venlafaxine did not inhibit ethoxyresorufin O-dealkylase (CYP1A2), tolbutamide 4-hydroxylase (CYP2C9) or testosterone 6 beta-hydroxylase (CYP3A4) activities at concentrations of up to 1 mM. CONCLUSIONS: It is concluded that venlafaxine has a low potential to inhibit the metabolism of substrates for CYP2D6 such as imipramine and desipramine compared with several of the most widely used SSRIs, as well as the metabolism of substrates for several of the other major human hepatic P450s. PMID- 9205823 TI - Endotoxin depresses hepatic cytochrome P450-mediated drug metabolism in women. AB - AIMS: In men, the inflammatory response to intravenous endotoxin depresses apparent oral clearances of antipyrine, hexobarbitone, and theophylline. The aim of this study was to investigate whether there might be gender differences in the regulation of hepatic cytochromes P450. METHODS: Experiments were carried out in seven healthy women volunteers (ages 19-51, median 22 years). Each woman received a cocktail of the three drugs on two occasions, once after a saline injection and again after endotoxin. RESULTS: Endotoxin injections, but not saline, caused the expected physiologic responses of inflammation including fever and increases in circulating tumor necrosis factor-alpha, interleukin-6, and C-reactive protein. When compared with the saline control studies, endotoxin significantly decreased clearances of all probes: antipyrine, 31% (95%CI 21%-41%); hexobarbitone, 20% (95%CI 10-31%); and theophylline, 20% (95%CI 10%-30%). The decreases were comparable with those found in the men previously studied (35%, 27%, and 22%, respectively). CONCLUSIONS: These data show that endotoxin-induced inflammation decreases hepatic cytochrome P450-mediated metabolism of selected probe drugs in women as it does in men. PMID- 9205824 TI - Comparison of the effects of a selective muscarinic receptor antagonist and hyoscine (scopolamine) on motion sickness, skin conductance and heart rate. AB - AIMS: Hyoscine (scopolamine), which is effective in the prophylaxis of motion sickness, shows similar binding affinities to all of the five known muscarinic receptor sub-types. The effectiveness of hyoscine was compared with zamifenacin (UK-76654), which binds selectively to the muscarinic M3 and m5 receptors. METHODS: Eighteen subjects received hyoscine hydrobromide 0.6 mg, zamifenacin 20 mg, or placebo (double-blind cross-over design). Sessions were 1 week apart and the drug (oral) was given 90 min prior to a motion sickness test. Motion sickness was elicited by cross-coupled stimulation on a turntable. The rotational velocity was incremented by 2 degrees s-1 every 30 s, and a sequence (seq) of eight head movements of 45 degrees was completed every 30 s. Motion tolerance was assessed as the number of sequences of head movement required to achieve moderate nausea. Pulse rate was recorded before and at 1 and 2 h after drug administration. Skin conductance activity in the frequency band 0.005-0.48 Hz, an index of sweat gland activity, was measured using Ag/AgCl electrodes on the palmar surfaces of fingers and across the forehead. RESULTS: Both zamifenacin and hyoscine produced an increase in tolerance to the motion challenge (P < 0.01) with no significant difference between the two drugs (5.0 +/- 1.6 vs 5.7 +/- 1.6 seqs. respectively, mean +/- s.e.mean). Compared with placebo or zamifenacin, pulse rate fell following hyoscine administration (9 beats min-1, P < 0.01). Skin conductance was reduced following hyoscine compared with zamifenacin or placebo (P < 0.001). CONCLUSIONS: These results suggest that compounds with selective M3 and/or m5 antagonism possess activity against motion sickness. Antagonism at these receptors may be the basis of the anti-motion sickness action of hyoscine. PMID- 9205826 TI - Consulting the oracle: a Delphi study to determine the content of a postgraduate distance learning course in therapeutics. AB - AIMS: To determine the content of a distance learning course in therapeutics for general practitioners (GPs). METHODS: This paper reports the results of a three round Delphi study. The respondent group comprised 21 GPs who were expert in their field. In the first round, the experts were interviewed to determine the knowledge, skills, competencies and attitudes required by GPs to prescribe effectively; the justification for, and scope of a distance learning course; and the preferred learning methods, format and methods of assessment. The first round generated 251 statements, which were collapsed into 108 statements covering thirteen domains. In the second and third rounds, a questionnaire was posted to the GPs. 95% responded to the second round questionnaire and agreed upon 86 statements, which were then collated into four domains. In the third round, 19 GPs rated each of the 86 statements on a five point scale. Consensus was reached for 99% of statements: 40 on the aims, design, format, organization and assessment of the course; and 45 on the knowledge and skills to be acquired by GPs who complete the course. RESULTS: The results revealed a consensus in favour of: modules at regular intervals some flexibility in meeting deadlines interaction among students for mutual support easy access to course tutors some face-to-face contact to complement distance learning material clear guidance on effective and safe prescribing emphasis on importance of 'people skills' CONCLUSIONS: The Delphi process can be used to determine the competencies required for continuing medical education in therapeutics. PMID- 9205825 TI - Compliance with a 2 day course of artemether-mefloquine in an area of highly multi-drug resistant Plasmodium falciparum malaria. AB - AIMS: Multi-drug resistant Plasmodium falciparum malaria is a rapidly increasing problem in the world, particularly Thailand. Practical antimalarial regimens which are highly effective against multi-drug resistant parasites with short-term course of administration are needed. In this study, we assessed the patient compliance of a short course regimen using artemether-mefloquine. METHODS: Clinical effectiveness (efficacy, tolerability and patient compliance) of a 2-day regimen of artemether-mefloquine was evaluated in 126 patients with acute uncomplicated falciparum malaria who were attending the two malaria clinics in an area of highly multi-drug resistant P. falciparum malaria (Thai-Myanmar border). Patients were treated with a single oral dose of 300 mg artemether on the day of attendance. Two additional doses of mefloquine were given for home treatment on the following day (750 and 500 mg after breakfast and lunch, respectively). RESULTS: The combination regimen was effective, with a cure rate of 92.6%. Based upon the concentrations of whole blood mefloquine on day-2, compliance for this 2 day regimen of artemether-mefloquine was 98.1% (full compliance 86.8%, partial compliance 11.3%, non-compliance 1.9%). CONCLUSIONS: We conclude that the 2 day regimen of artemether-mefloquine is, at present, a good alternative regimen for the treatment of uncomplicated multi-drug resistant falciparum malaria. PMID- 9205827 TI - Hospitalization for serious blood and skin disorders following co-trimoxazole. AB - AIMS: To quantify the risk of serious blood and skin disorders requiring hospitalization among otherwise healthy users of co-trimoxazole. METHODS: We conducted a population-based cohort study at Group Health Cooperative of Puget Sound (GHC). RESULTS: During the years 1987 to 1993 we found six cases of co trimoxazole-associated blood disorders and three cases of co-trimoxazole associated skin disorders yielding risks of 5.6/100,000 (95% CI 2.6-12.2) and 2.8/100,000 (95% CI 0.9-8.2) respectively. In all cases found there was prompt recovery after discontinuation of co-trimoxazole. We found no cases of toxic epidermal necrolysis. CONCLUSIONS: We conclude that the risk of blood and skin disorders associated with the use of co-trimoxazole leading to hospitalization is low. PMID- 9205828 TI - The pathophysiological mechanism of fluid retention in advanced cancer patients treated with docetaxel, but not receiving corticosteroid comedication. AB - AIMS: Fluid retention is a phenomenon associated with taxoids. The principal objective of this study was to investigate the pathophysiological mechanism of docetaxel-induced fluid retention in advanced cancer patients. METHODS: Docetaxel was administered as a 1 h intravenous infusion every 3 weeks, for at least 4-6 consecutive cycles, to patients with advanced breast (n = 21) or ovarian (n = 3) carcinoma, who had received previous chemotherapy, 21 for advanced disease. Phase II clinical trials have shown that 5 day corticosteroid comedication, starting 1 day before docetaxel infusion, significantly reduces the incidence and severity of fluid retention. This prophylactic corticosteroid regimen is currently recommended for patients receiving docetaxel but was not permitted in this study because of its possible interference with the underlying pathophysiology of the fluid retention. RESULTS: Fluid retention occurred in 21 of the 24 patients but was mainly mild to moderate, with only five patients experiencing severe fluid retention. Eighteen patients received symptomatic flavonoid treatment, commonly prescribed after the last cycle. Specific investigations for fluid retention confirmed a relationship between cumulative docetaxel dose and development of fluid retention. Capillary filtration test analysis showed a two-step process for fluid retention generation, with progressive congestion of the interstitial space by proteins and water starting between the second and the fourth cycle, followed by insufficient lymphatic drainage. CONCLUSIONS: A vascular protector such as micronized diosmine hesperidine with recommended corticosteroid premedication and benzopyrones may be useful in preventing and treating docetaxel-induced fluid retention. PMID- 9205829 TI - Association between CYP2C19 genotype and proguanil oxidative polymorphism. AB - AIMS: To examine the relationship between proguanil metabolism and the number of mutations in CYP2C19 by comparing the CYP2C19 genotype and proguanil phenotype of 10 subjects. METHODS: Partial clearance and urinary metabolic ratio data were obtained from a previous study of 10 subjects [5]. Analysis of CYP2C19 genotypes was performed using PCR amplification followed by restriction endonuclease digestion of genomic DNA from a blood sample. RESULTS: The intrinsic partial clearance of PG to CG ranged from 0.41-10.11 h-1, and was related to the number of functional CYP2C19 alleles present. Genotypic PMs had metabolic ratios > 13, while genotypic heterozygote EMs had metabolic ratios < 9. CONCLUSIONS: Proguanil may be a suitable phenotyping probe for the CYP2C19 genetic polymorphism, however the exact antimode of the urinary metabolic ratio chosen to separate poor and extensive metabolisers needs further investigation. PMID- 9205830 TI - Lack of effect of eprosartan on the single dose pharmacokinetics of orally administered digoxin in healthy male volunteers. AB - AIMS: To study the effect of eprosartan, a nonbiphenyl tetrazole angiotensin II receptor antagonist, on digoxin pharmacokinetics in a randomized, open-label, two period, period balanced crossover study in 12 healthy men. METHODS: Each subject received a single 0.6 mg oral dose of digoxin (Lanoxicaps 0.2 mg/capsule, Glaxo Wellcome) alone or following 4 days of dosing with eprosartan 200 mg orally every 12 h. Each study period was separated by a 14 day washout interval. Serial blood samples were obtained for up to 96 h after each digoxin dose for determination of digoxin pharmacokinetics. The effect of eprosartan on digoxin pharmacokinetics was assessed through an equivalence-type approach using AUC(0, t') as the primary endpoint. RESULTS: For AUC(0, t'), the ratio of digoxin+eprosartan: digoxin alone was 0.99 with a 90% confidence interval (CI) of [0.90, 1.09]. For Cmax, the ratio was 1.00 with a 90% CI of [0.86, 1.17]. tmax was similar for both regimens. Both regimens were safe and well tolerated. CONCLUSIONS: Based on AUC and Cmax data, it can be concluded that eprosartan has no effect on the pharmacokinetics of a single oral dose of digoxin. PMID- 9205831 TI - Neurological, cardiovascular and metabolic effects of mefloquine in healthy volunteers: a double-blind, placebo-controlled trial. PMID- 9205832 TI - Altered neurotransmission and signal transduction: targets for nicergoline treatment. AB - Although we do not yet have a complete understanding of the molecular mechanisms responsible for the pathophysiology of Alzheimer's disease, it is clear that a drug such as nicergoline, which may have both acute and longer-term effects on the disease, could offer a beneficial and worthwhile approach to treatment. PMID- 9205833 TI - Protein kinase C in synaptic plasticity: a molecular target in the treatment of cognitive disorders. PMID- 9205834 TI - Relations between symptomatology and brain function in dementias: double-blind, placebo-controlled, clinical and EEG/ERP mapping studies with nicergoline. PMID- 9205835 TI - Nicergoline: parallel evolution of clinical trial methodology and drug development in dementias. PMID- 9205836 TI - Prediction of gene expression specificity by promoter sequence patterns. AB - We present here a heuristic method toward predicting the expression specificity in the transcriptional process, which is known to be regulated in large part by promoter sequences, by observing the appearance of conserved sequence patterns in a group of known promoters, such as for housekeeping or tissue-specific genes. Statistically conserved patterns were automatically extracted from a set of unaligned sequences up to 200 bp upstream of the transcription initiation site, by a standard procedure using the Markov chain and binomial distribution models. Furthermore, to obtain signal sequences of optimal lengths we devised a method that combines the multiple alignment and the analysis of the information content (or relative entropy). Groups of related promoters were compiled from the EPD eukaryotic promoter database and the EMBL nucleic acid sequence database. Each promoter was examined for its specificity by linear discriminant analysis to test the validity of the extracted patterns. Our method could correctly discriminate 77.6% of the housekeeping gene promoters and 62.9% of the liver promoters. PMID- 9205837 TI - Construction of a contiguous 874-kb sequence of the Escherichia coli -K12 genome corresponding to 50.0-68.8 min on the linkage map and analysis of its sequence features. AB - The contiguous 874.423 base pair sequence corresponding to the 50.0-68.8 min region on the genetic map of the Escherichia coli K-12 (W3110) was constructed by the determination of DNA sequences in the 50.0-57.9 min region (360 kb) and two large (100 kb in all) and five short gaps in the 57.9-68.8 min region whose sequences had been registered in the DNA databases. We analyzed its sequence features and found that this region contained at least 894 potential open reading frames (ORFs), of which 346 (38.7%) were previously reported, 158 (17.7%) were homologous to other known genes, 232 (26.0%) were identical or similar to hypothetical genes registered in databases, and the remaining 158 (17.7%) showed no significant similarity to any other genes. A homology search of the ORFs also identified several new gene clusters. Those include two clusters of fimbrial genes, a gene cluster of three genes encoding homologues of the human long chain fatty acid degradation enzyme complex in the mitochondrial membrane, a cluster of at least nine genes involved in the utilization of ethanolamine, a cluster of the secondary set of 11 hyc genes participating in the formate hydrogenlyase reaction and a cluster of five genes coding for the homologues of degradation enzymes for aromatic hydrocarbons in Pseudomonas putida. We also noted a variety of novel genes, including two ORFs, which were homologous to the putative genes encoding xanthine dehydrogenase in the fly and a protein responsible for axonal guidance and outgrowth of the rat, mouse and nematode. An isoleucine tRNA gene, designated ileY, was also newly identified at 60.0 min. PMID- 9205838 TI - Mechanism of anucleate cell production in the oriC-deleted mutants of Bacillus subtilis. AB - We constructed oriC-deleted mutants of Bacillus subtilis by integrating the minimal replication region of plasmid pLS32 into the proA (115 degrees), spoIIIJ (360 degrees) and thrS (256 degrees) loci of the chromosome, respectively. All three mutants produced anucleate cells and the DNA/protein ratio was lower than that of the wild-type strain when grown in nutrient broth. However, when grown in minimal-glucose medium, the frequency of anucleate cells was reduced in all of them and the DNA/protein ratio was restored to normal. Especially, the oriC deleted mutant in which the plasmid was integrated near oriC produced almost no anucleate cell. These results indicate that initiation frequency of chromosome replication from the integrated plasmid origin were reduced disproportionately to cell mass increase in rich medium, which in turn disrupted coordination between DNA replication cycle and cell division cycle. The locations of the plasmid origin relative to the natural oriC locus affected the production of anucleate cell remarkably, suggesting that partition mechanism of chromosome was also impaired by the translocation of its replication origin. PMID- 9205839 TI - Yeast artificial chromosome clones of rice chromosome 2 ordered using DNA markers. AB - Yeast artificial chromosome (YAC) clones were ordered for the physical mapping of rice chromosome 2, the last of the 12 rice chromosomes to be assigned YACs by the Rice Genome Research Program. A total of 128 restriction fragment length polymorphism markers and 4 sequence-tagged site (STS) markers located on our high density genetic map were used for YAC clone landing. By colony/Southern hybridization and polymerase chain reaction screening, a total of 239 individual YACs were selected from our YAC library of 6934 clones covering six genome equivalents. The YACs located on the corresponding marker positions in the linkage map formed 43 contigs and islands and were estimated to encompass about 50% of the length of rice chromosome 2. PMID- 9205840 TI - Ordered YAC clone contigs assigned to rice chromosomes 3 and 11. AB - Yeast artificial chromosome (YAC) clones were arranged on the positions of restriction fragment length polymorphism (RFLP) and sequence-tagged site (STS) markers already mapped on the high-resolution genetic maps of rice chromosomes 3 and 11. From a total of 416 and 242 YAC clones selected by colony/Southern hybridization and polymerase chain reaction (PCR) analysis, 238 and 135 YAC clones were located on chromosomes 3 and 11, respectively. For chromosomes 3 and 11, 24 YAC contigs and islands with total coverage of about 46% and 12 contigs and islands with coverage of about 40%, respectively, were assigned. Although many DNA fragments of multiple copy marker sequences could not be mapped to their original locations on the genetic map by Southern hybridization because of a lack of RFLP, the physical mapping of YAC clones could often assign specific locations of such multiple copy sequences on the genome. The information provided here on contig formation and similar sequence distribution revealed by ordering YAC clones will help to unravel the genome organization of rice as well as being useful in isolation of genes by map-based cloning. PMID- 9205841 TI - Prediction of the coding sequences of unidentified human genes. VII. The complete sequences of 100 new cDNA clones from brain which can code for large proteins in vitro. AB - In this series of projects of sequencing human cDNA clones which correspond to relatively long transcripts, we newly determined the entire sequences of 100 cDNA clones which were screened on the basis of the potentiality of coding for large proteins in vitro. The cDNA libraries used were the fractions with average insert sizes from 5.3 to 7.0 kb of the size-fractionated cDNA libraries from human brain. The randomly sampled clones were single-pass sequenced from both the ends to select clones that are not registered in the public database. Then their protein-coding potentialities were examined by an in vitro transcription/translation system, and the clones that generated proteins larger than 60 kDa were entirely sequenced. Each clone gave a distinct open reading frame (ORF), and the length of the ORF was roughly coincident with the approximate molecular mass of the in vitro product estimated from its mobility on SDS-polyacrylamide gel electrophoresis. The average size of the cDNA clones sequenced was 6.1 kb, and that of the ORFs corresponded to 1200 amino acid residues. By computer-assisted analysis of the sequences with DNA and protein motif databases (GenBank and PROSITE databases), the functions of at least 73% of the gene products could be anticipated, and 88% of them (the products of 64 clones) were assigned to the functional categories of proteins relating to cell signaling/communication, nucleic acid managing, and cell structure/motility. The expression profiles in a variety of tissues and chromosomal locations of the sequenced clones have been determined. According to the expression spectra, approximately 11 genes appeared to be predominantly expressed in brain. Most of the remaining genes were categorized into one of the following classes: either the expression occurs in a limited number of tissues (31 genes) or the expression occurs ubiquitously in all but a few tissues (47 genes). PMID- 9205842 TI - Characterization of the promoter region, first ten exons and nine intron-exon boundaries of the DNA-dependent protein kinase catalytic subunit gene, DNA-PKcs (XRCC7). AB - The gene, DNAPKcs (XRCC7), for the human DNA-dependent protein kinase catalytic subunit (DNA-PKcs) is a strong candidate that complements a severe combined immunodeficiency (scid) and hypersensitivity to ionizing radiation in mice. We constructed a cosmid library from a previously identified, X RCC7-covering YAC clone (943G4). From the library, we isolated three cosmid clones containing the 5'-region of XRCC7. Sequence analysis with primer walking on a 6.3-kb segment of these cosmids identified the promoter region, the first ten exons and nine intron exon boundaries of XRCC7. The promoter region contains several potential Sp1 protein-binding sites and a high G+C content but no TATA or CCAAT boxes. These findings are consistent with the TATA-less housekeeping gene promoter and provides the basis for transcriptional regulatory studies. Since nine other exons spanning an 8-kb segment are already known, a total of 19 exons in the gene have been identified. The cosmids isolated and the primer sets designed in the present study are useful for mutation analysis in patients with a SCID phenotype. PMID- 9205843 TI - Sequence analysis of the 36-kb region between gntZ and trnY genes of Bacillus subtilis genome. AB - Within the framework of an international Bacillus subtilis genome sequencing project, we have determined a 36-kb sequence covering the region between the gntZ and trnY genes. In addition to five genes sequenced and characterized previously, 27 putative protein coding sequences (open reading frame; ORF) were identified. A homology search for the newly identified ORFs revealed that six of them had similarities to known proteins. It is notable that new ORFs belonging to response regulator aspartate phosphatase (Rap) and its regulator (Phr) families, and response regulator and sensory kinase families of two-component signal transduction systems have been identified. Furthermore, we found that some 180-bp non-coding sequence, that might be an remnant of an ancient IS element, is preserved in at least five loci of the B. subtilis genome. PMID- 9205844 TI - Compilation of all genes encoding two-component phosphotransfer signal transducers in the genome of Escherichia coli. AB - Bacteria have devised sophisticated His-Asp phosphorelay signaling systems for eliciting a variety of adaptive responses to their environment, which are generally referred to as the "two-component regulatory system." The widespread occurrence of the His-Asp phosphorelay signaling in both prokaryotes and eukaryotes implies that it is a powerful device for a wide variety of adaptive responses of cells to their environment. The two-component signal transducers contain one or more of three common and characteristic phosphotransfer signaling domains, named the "transmitter, receiver, and histidine-containing phosphotransfer (HPt) domains." The recently determined entire genomic sequence of Escherichia coli allowed us to compile systematically a complete list of genes encoding such two-component signal transduction proteins. The results of such an effort, made in this study, revealed that at least 62 open reading frames (ORFs) were identified as putative members of the two-component signal transducers in this single species. Among them, 32 were identified as response regulator and 23 were identified as orthodox sensory kinases. In addition, E. coli has five hybrid sensory kinases. The precise location of each ORF was mapped on a physical map of the entire E. coli genome. All of these ORFs were then compiled and annotated extensively. PMID- 9205845 TI - Construction of a contiguous 874-kb sequence of the Escherichia coli K-12 genome corresponding to the 50.0-68.8 min on the linkage map and analysis of its sequence features (supplement). PMID- 9205846 TI - Future directions in the development of new antitubercular drugs. Where do we go from here? AB - The global resurgence of tuberculosis and rampant drug resistance have rekindled the need for and interest in the development of new antitubercular drugs. Delineation of the possible drug targets furnished by the various mycobacterial cell components might result in rational approaches to the development of such agents. In the future, molecular genetics might help both in the bioengineering and rapid screening of the activity of new compound. Collaboration is anticipated between the pharmaceutical industry and academic institutions in these areas. PMID- 9205847 TI - When, and when not, to use digoxin in the elderly. AB - Digitalis has been widely used in the treatment of cardiac disease for more than 200 years. The present article reviews the current role of digitalis in the management of heart failure and atrial fibrillation (AF) in light of recent study findings. Generally, first-line therapy for the management of heart failure due to systolic dysfunction should include an ACE inhibitor and a diuretic. In patients who remain symptomatic despite the use of these drugs, the addition of digoxin should be considered. Because digoxin has been shown to reduce the number of hospital admissions attributable to worsening heart failure, more liberal use of digoxin in the management of heart failure may be justified. Digoxin may be adequate as monotherapy for ventricular rate control in patients with chronic AF, particularly in sedentary and elderly patients. A beta-blocker or calcium antagonist (either alone or in combination with digoxin) is indicated when digoxin is ineffective for ventricular rate control. Digoxin is ineffective in restoring sinus rhythm, preventing paroxysms or controlling rate in paroxysmal AF. The elderly are at an increased risk of digoxin toxicity. Low dosages of digoxin appear to be effective in the treatment of heart failure due to systolic dysfunction and may reduce the incidence of digitalis toxicity in these patients. In elderly patients with AF and inadequate rate control who are receiving digitalis monotherapy, adding another atrioventricular nodal blocking drug may be more appropriate than increasing the digoxin dose, in order to avoid toxic digoxin levels. PMID- 9205849 TI - Optimal treatment of heart failure in the elderly. AB - Heart failure is a common condition in the elderly and one that is likely to become more prevalent as the population ages. Many drugs have been developed for the treatment of heart failure, but because clinical trials of these agents have often not included elderly patients their results need to be extrapolated from younger to older patients. Age-related physiological changes that affect how the available treatments are used occur in many organ systems. Effective management strategies can be implemented in elderly patients as well as in younger age groups, and these can improve both functional status and quality of life as well as reducing hospital admission and improving survival. This article reviews the physiological changes that occur in the elderly and the treatment approach that can be taken in elderly patients with heart failure. PMID- 9205848 TI - Angiotensin II receptor antagonists. Potential in elderly patients with cardiovascular disease. AB - Raised blood pressure in the elderly is not a normal consequences of aging, but is a major risk factor for cardiovascular disease. Cardiac and cerebrovascular disease account for > 50% of deaths among people aged > 65 years. Because the percentage of elderly people in most populations is rising, blood pressure control in this group is becoming increasingly important. Several large intervention studies in the elderly have demonstrated that antihypertensive medication reduces cardiovascular morbidity and mortality. In addition, the absolute benefits of blood pressure reduction are higher in elderly compared with younger patients. ACE inhibitors are effective and well tolerated in the treatment of hypertension in the elderly. Their success led to interest in alternative ways of blocking the renin angiotensin system, and the subsequent development of angiotensin II (AII) receptor antagonists. Losartan was the first drug in this class to become commercially available. Since then, valsartan has been launched in some markets and others are likely to be launched in the near future. Losartan is effective in the treatment of essential hypertension and has a low incidence of adverse effects. First-dose hypotension is very uncommon and, at the present time, cough does not appear to be an adverse effect of these drugs, although long term tolerability studies are needed to confirm this. Angioedema, a rare but life-threatening adverse effect of ACE inhibitors, has also been associated with losartan. Current data suggest that All receptor antagonists are effective in elderly hypertensive patients, although further data are needed to confirm these findings. At present, All receptor antagonists are likely to be used in hypertensive patients who are intolerant of ACE inhibitors, although this may change with the availability of long term tolerability and clinical outcomes data. PMID- 9205851 TI - Age-related changes in cardiac physiology. Can they be postponed or treated by drugs? AB - The basic mechanisms that cause aging are still poorly understood. Longitudinal prospective population studies using noninvasive examination techniques have improved our ability to differentiate between aging and disease. This review describes some general morphological and functional aging-related changes of the heart that have clinical relevance, and considers the possibility of drug treatment for the manifestations of aging per se. Digitalis has not been shown to improve the aging-related decline in myocardial strength and contractility. During aging, heart tissue stiffens and the speed and extent of diastolic filling decline. The latter is a limiting functional factor, particularly during increases in heart rate. Lowering peripheral vascular resistance, which is often increased in older people, might indirectly improve heart function. However, no drug has been shown to improve myocardial strength or lower tissue stiffness via a direct effect on the heart. It has been claimed, however, that calcium antagonists might improve diastolic filling. Morphological changes during aging are dominated by some left ventricular wall and septal hypertrophy, and left atrial and ventricular widening. Recent findings have suggested that angiotensin II might act as a growth stimulating factor, promoting cardiac hypertrophy. This has led to speculation that ACE inhibitors might contribute to the restructuring of the heart, not only in hypertension but also in patients with the common combination of slightly elevated blood pressure and aging-related myocardial hypertrophy. At present, it appears that improving exogenous factors (e.g. lifestyle, living circumstances and access to adequate medical care) offers greater opportunities for postponing cardiac aging than drugs that directly interfere with the physiological aging of the heart. PMID- 9205853 TI - The cytogenetics of lipomatous tumours. AB - Precise classification of lipomatous tumours sometimes presents a challenge to even the most experienced pathologist. Because the different types of lipomatous tumour exhibit significantly different biological behaviours, it is of utmost importance that the correct diagnosis be made. In morphologically difficult cases, identification of characteristic cytogenetic aberrations that correlate with histological subtype can facilitate the correct diagnosis. PMID- 9205854 TI - Hepatitis C virus liver disease in women infected with contaminated anti-D immunoglobulin. AB - Screening for hepatitis C virus (HCV) infection is carried out by detection of antibodies to the virus (enzyme-linked immunosorbent assay (ELISA) and recombinant immunoblot assay (RIBA)) with confirmation by identification of HCV RNA genome in serum (polymerase chain reaction (PCR)). We describe the histological features on liver biopsy in 88 women with chronic HCV infection (serum positive on ELISA, RIBA and PCR) acquired from virus contaminated anti-D immunoglobulin. For the majority of these patients the time interval from virus infection to presentation was between 17 and 18 years. We separately assessed necroinflammatory disease activity and architectural features on liver biopsy and applied a scoring system which permitted semi-quantitative documentation of abnormal features. Only three women showed liver biopsies within normal limits (+/-focal steatosis). The remaining 85 cases showed a predominantly mild or moderate degree of disease activity with interface hepatitis (56.8% of cases), spotty necrosis, apoptosis and focal inflammation (88.6% of cases) and portal inflammation (90.9% of cases). Confluent necrosis was an uncommon finding (2.3% of cases). Assessment of architectural features showed normal appearance in 35.2% of biopsies. The predominant architectural abnormality noted was portal tract fibrosis. Ten per cent of cases, however, showed significant fibrous band and/or nodule formation. PMID- 9205850 TI - Management of lipid disorders in the elderly. AB - Cardiovascular disease has been inseparable from aging in developed societies and, as a result, it is the commonest cause of mortality in elderly populations. Atherosclerosis is associated with the progressive vascular accumulation of cholesterol-laden lipoproteins, and is linearly associated with the plasma level of low density lipoprotein (LDL) cholesterol. Clinical trials in patients aged < 65 years have conclusively shown that treatment of hypercholesterolaemia decreases the incidence of cardiovascular events and total mortality. However, few conclusive data are available regarding the treatment of hypercholesterolaemia in elderly patients. Extrapolation from clinical trials suggests that lipid lowering treatment in well selected elderly patients is effective in preventing cardiovascular events and is an efficient use of healthcare resources. In addition to cholesterol, high triglyceride and low high density lipoprotein levels appear to be significant predictors of coronary artery disease in elderly patients. We do not advocate the indiscriminate screening of healthy elderly patients who have no other cardiovascular risk factors, because the marginal overall benefits are probably small and the costs of widespread screening and treatment high. On the other hand, chronological age itself cannot be considered a barrier to the screening and treatment of patients who have a good quality of life but have other cardiovascular risk factors and/or definite cardiovascular disease. Subgroup analysis of major clinical trials suggests that the aims of treatment should be to lower the LDL cholesterol level to 3.2 mmol/L (125 mg/dl), or the total cholesterol level to 5.2 mmol/L (200 mg/dl). Occasionally, multiple drug therapy is required to achieve this target, but statins (HMG-CoA reductase inhibitors) are the most commonly used first-line agents. With aggressive lowering of plasma lipid levels in this way, a reduction in clinical events is paralleled by regression of atheroma detectable by angiography, and an improvement in endothelial function. Global reduction of risk factors in elderly patients should always be undertaken, including dietary therapy, weight reduction in viscerally obese patients, postmenopausal estrogen replacement, smoking cessation, treatment of hypertension and control of diabetes mellitus. A secondary cause of dyslipidaemia should also be sought. The role of antioxidants is still not clear, but they are probably of little benefit in elderly patients. With the widespread use of effective, well tolerated treatments for lipid disorders in younger patients, significant improvements have already been attained in the morbidity and mortality associated with coronary artery disease. Since the current life expectancy at age 65 years is nearly 20 years in most Western countries, secondary prevention may increase the quality of life and the independent lifespan, even if eventual mortality is not delayed. PMID- 9205852 TI - Mitoxantrone. A review of its pharmacology and clinical efficacy in the management of hormone-resistant advanced prostate cancer. AB - The antineoplastic agent mitoxantrone in combination with a corticosteroid (either prednisone or hydrocortisone) has shown clinical efficacy as palliative treatment for a proportion of patients (about 35 to 40%) with hormone-resistant advanced prostate cancer, a disease which predominantly affects elderly men and for which few systemic treatment options are available. Palliative end-points including pain relief, decreased analgesic use and reduced prostate specific antigen levels (a marker of tumour response) are reached in a greater percentage of patients receiving combination therapy than corticosteroid alone. In addition, there are generally greater improvements in quality-of-life parameters in mitoxantrone recipients. However, combined treatment offers no survival advantage over corticosteroid monotherapy. Neutropenia is the most common toxicity associated with mitoxantone therapy and may necessitate dosage reduction in some patients. Otherwise, mitoxantrone generally has a more favourable tolerability profile than has been established for other cytotoxic agents such as doxorubicin with regard to acute adverse events (e.g. nausea/vomiting, anorexia, constipation, alopecia, malaise/ fatigue, oedema) and cardiac toxicity. In conclusion, administration of mitoxantrone plus a corticosteroid can provide palliation for some elderly patients with hormone-resistant advanced prostate cancer, and is thus a valuable first-line treatment for this indication. PMID- 9205855 TI - Immunohistochemical staining of hepatocellular carcinoma with monoclonal antibody against inhibin. AB - Inhibin is a peptide hormone produced by ovarian granulosa cells. During a recent study investigating the immunohistochemical staining of ovarian granulosa cell tumours and other neoplasms with an anti-inhibin monoclonal antibody, we identified strong cytoplasmic staining of hepatocytes. In the present study we investigated the immunostaining of hepatocellular carcinoma and other neoplasms involving the liver with anti-inhibin to determine whether the antibody may be of value in the differential diagnosis of hepatic neoplasms. Immunostaining for alpha-fetoprotein was also performed. With anti-inhibin there was positive, generally strong, cytoplasmic staining of 17 of 19 cases of hepatocellular carcinoma, including the pleomorphic and glandular variants. There was positive staining of six of 20 cases of adenocarcinoma. In these, positive staining was generally focal, of weak intensity and involved the luminal surface of neoplastic glands. There was no staining of five cases of neuroendocrine tumour. There was positive staining for alpha-fetoprotein in 13 of 19 cases of hepatocellular carcinoma and in two of 20 cases of adenocarcinoma but no staining of neuroendocrine tumours. Immunostaining with anti-inhibin antibody may be of value in the differentiation of hepatocellular carcinoma from other neoplasms involving the liver. The antibody is a more sensitive, but less specific, immunohistochemical marker for hepatocellular carcinoma than is alpha fetoprotein. PMID- 9205856 TI - Prognostic comparison of three classifications for medullary carcinomas of the breast. AB - The aim of this study was to make prognostic comparisons between the modified scheme of Pedersen et al. the definitions of Tavassoli and the Ridolfi criteria for medullary carcinomas. Sixty breast carcinomas primarily diagnosed as medullary carcinomas were reclassified into typical medullary carcinoma (TMC), atypical medullary carcinoma (AMC) and non-medullary carcinoma (NMC) according to the three classifications. The Ridolfi classification proved to be superior to the two other schemes in discriminating survival differences between the three groups TMC, AMC and NMC. All 13 patients with TMC are still alive indicating an excellent prognosis, while 29% and 39% of the 47 patients in the AMC and NMC category, respectively, have died of their disease. In the simplified system of Pedersen et al. the survival at 10 years for TMC patients decreased to 75% and no significant survival difference between the three groups could be demonstrated. As the prognosis for AMC proved to be worse compared to TMC and in fact was similar to NMC with values of 43% at 10 years in the Ridolfi classification, we find no reasons to maintain this category. We conclude that as long as no alternative and more easily applicable diagnostic method exists, pathologists should still apply the Ridolfi criteria on these tumours with medullary features leaving two diagnostic possibilities: TMC or NMC (i.e. poorly differentiated ductal carcinoma). Only lesions that fulfil all six criteria without any doubt should be diagnosed as TMC, thus avoiding overdiagnosis and a resulting risk of undertreatment. PMID- 9205857 TI - Epithelial hyperplasia of usual type expresses both S100-alpha and S100-beta in a heterogeneous pattern but ductal carcinoma in situ can express only S100-alpha in a monotonous pattern. AB - Immunohistochemical identification of myoepithelial cells using alpha-smooth muscle actin provides little information about the nature of solid or quasi-solid portions of epithelial hyperplasia and ductal carcinoma in situ (DCIS) because actin-rich myoepithelial cells are usually demonstrated only in the stromal epithelial junction of both lesions. We studied the differential distribution of alpha-subunit (S100-alpha) and beta-subunit (S100-beta) of S100 protein in actin negative areas of usual epithelial hyperplasia and DCIS by employing the streptavidin method with monospecific rabbit antibodies against each subunit. All usual epithelial hyperplasias (n = 17) were composed of heterogeneous epithelial cell types; cells expressing S100-alpha and/or S100-beta were intermingled with non-expressing cells, resulting in a mosaic-like pattern. On the contrary, DCIS (n = 32) uniformly lacked immunoreactive S100-beta; S100-alpha was diffusely expressed in 24 (68.8%) DCIS (three solid/comedo, 13 cribriform, four endocrine, one micropapillary, three papillary variants) and negative in the remaining eight (31.2%) DCIS (one cribriform, two micropapillary, four papillary and one apocrine variants). In conclusion, in contrast to usual epithelial hyperplasia that expresses both S100-alpha and S100-beta in a heterogeneous pattern, DCIS can express only S100-alpha in a monotonous pattern, possibly signifying unidirectional differentiation toward secretory glandular epithelium. PMID- 9205858 TI - Poorly differentiated myoepithelial cell rich carcinoma of the breast. AB - Three unusual cases of invasive breast carcinoma are reported, each comprising a dual malignant cellular proliferation consisting of 'ordinary' epithelial cells as well as myoepithelial cells haphazardly intermingled. The cases displayed features which did not match any of the four main types of invasive malignant breast tumours containing myoepithelial elements, i.e. adenomyoepithelioma, adenoid cystic carcinoma, pure myoepithelioma and low-grade adenosquamous carcinoma. The designation of 'poorly differentiated myoepithelial cell rich carcinoma' (PDMC) is proposed for these tumours. PMID- 9205860 TI - Immunodetection of androgen receptor in human urinary bladder cancer. AB - We investigated the expression of androgen receptor (AR) protein in transitional cell carcinoma of human urinary bladder in paraffin-embedded sections of tumours obtained from nine patients with urinary bladder cancer treated by radical cystectomy. In addition, immunoblotting of AR was also performed on selected samples. Nuclear immunoreactivity of AR was found in seven of the nine urinary bladder cancers studied. AR showed variable staining intensity within a tumour. In the immunoblots, a 110 kDa AR signal was seen with anti-AR antibody, and faint bands of 90 and 60 kDa were also observed. Immunohistochemistry of p53 and c-erbB 2 was also carried out and compared with the distribution of AR. The high frequency of AR expression suggests a role for androgens in transitional cell carcinoma of human urinary bladder. PMID- 9205859 TI - Expression of p53, mdm2, p21/waf1 and bcl-2 proteins in thymomas. AB - We have investigated the immunohistochemical expression of p53, mdm2, p21/waf1 and bcl-2 proteins in 31 thymic epithelial tumours comprising five medullary thymomas (MDT), four mixed thymomas (MT), 12 cortical thymomas (CT), eight predominately cortical thymomas (PCT) and two well-differentiated thymic carcinomas (WDTC). We have found p53, mdm2, p21 and bcl-2 protein expression in 25/31, 8/31, 5/31 and 10/31 thymic epithelial tumours, respectively. Coexpression of p53 and mdm2 proteins was found in eight cases (three CT, four PCT and one WDTC). Five of them were also p21 positive and three p21 negative. Discordant p53+/mdm2-/p21- protein expression was found in 19 cases (three MDT, three MT, nine CT, three PCT and one WDTC). Mdm2 and p21 proteins were not expressed in the absence of p53 protein. Coexpression of bcl-2 and p53 proteins was found in seven cases (three MDT, three MT and one WDTC). Eighteen cases were p53+/bcl-2- (10 CT, seven PCT and one WDTC) and three cases (two MDT and one MT) were bcl-2+/p53-. Our findings indicate that in thymomas, p53 expression is more frequently associated with cortical histotypes while bcl-2 expression is strongly associated with medullary and mixed histotypes. In addition, there is an inverse correlation between p53 and bcl-2 protein expression in thymomas. Coexpression of p53/mdm2/p21 proteins may reflect thymomas with wild-type (wt), p53 gene since mdm2 and p21 proteins are inducible by wt p53 gene. However, in view of previous findings that p53 mutation is an early event in thymomas, the possibility of p53 gene mutation with p53-independent mdm2 and p21 expression should be considered in these cases. Discordant p53+/mdm2-/p21- protein expression may represent thymomas with p53 gene mutations unable to activate expression of mdm2 and p21 proteins. PMID- 9205861 TI - Umbilical cord 'pseudo-vasculitis' following second trimester fetal death: a clinicopathological and immunohistochemical study of 13 cases. AB - Amniotic fluid bacterial infection is an occasional cause of second trimester septic abortion. We describe an autolysis-related histological artifact, umbilical cord 'pseudo-vasculitis', which can erroneously implicate amniotic bacterial infection in fetal death. Clinicopathological features of 13 second trimester fetal deaths with umbilical cord pseudo-vasculitis are reported. In four cases (31%), an incorrect pathological diagnosis of umbilical vasculitis had initially been rendered. Umbilical cords from five cases of pseudo-vasculitis and one comparison fetus (18-week septic abortion with true umbilical vasculitis), were studied with chloroacetate esterase and with immunohistochemical staining for myeloperoxidase, muscle-specific actin (HHF35) and smooth muscle actin. Histologically, umbilical pseudo-vasculitis exhibited numerous small, rounded, degenerating cells with irregular, multilobed nuclei (closely resembling neutrophils) located within the umbilical vessel wall. Immunohistochemical studies demonstrated that all cells resembling neutrophils were of smooth muscle origin. Moderate to severe fetal autolysis was present in all cases of umbilical pseudo-vasculitis, suggesting that this finding represents autolysis of umbilical vascular smooth muscle secondary to post moderate fetal retention. Vascular smooth muscle in other fetal and placental locations did not demonstrate the finding, suggesting that this striking degenerative artifact of smooth muscle is restricted to the umbilical cord. PMID- 9205862 TI - Multifocal alveolar hyperplasia associated with lymphangioleiomyomatosis in tuberous sclerosis. AB - Multifocal alveolar hyperplasia associated with pulmonary lymphangioleiomyomatosis is reported in a 21-year-old woman with tuberous sclerosis. Beside the cystic lesions of lymphangioleiomyomatosis, the tomography showed nodules up to 8 mm in both upper lobes. A proliferation of type II pneumonocytes and Clara cells lining the alveolar walls in an adenoma-like pattern was observed. Nuclear atypia, mitoses and necrosis were not observed, providing evidence against multicentric bronchioloalveolar carcinoma or micronodular atypical alveolar adenomatous hyperplasia. Whereas the lymphangioleiomyomatosis lesions showed strong positivity for HMB45 and expressed oestrogen and progesterone receptors, the alveolar hyperplasia was negative for these markers as it was for carcinoembryonic antigen, p53 and MIB1 antibodies. Multifocal alveolar hyperplasia in tuberous sclerosis is probably a benign hamartomatous lesion in our case without progression on a 2-year follow-up. Its histogenesis is unknown, but is possibly related to chromosome instability. PMID- 9205863 TI - Meningeal melanocytoma: case report and review of the literature. AB - We report a case of meningeal melanocytoma in the thoracic spinal cord of a 44 year-old woman and review previously documented cases. Our patient experienced numbness and tingling in her left leg for 8 years, and low back pains with intermittent claudication for the previous 2 months. A histologically benign 20 mm tumour was totally resected. Radiation therapy was not given. The tumour showed the histological, immunohistochemical and ultrastructural features of a meningeal melanocytoma. The patient is alive without recurrence 4.5 years after surgery. PMID- 9205864 TI - Systemic mastocytosis following a malignant ovarian germ cell tumour. AB - Cases of mediastinal germ cell tumours associated with haematological disorders (two cases of systemic mastocytosis included) have been reported previously. This combination is more frequent than would be expected by chance alone. We report the case of a 30-year-old woman, who presented with a systemic mastocytosis following a malignant ovarian germ cell tumour which was treated by chemo- and radiotherapy. The patient predominantly complained of skeletal pains, which led to an erroneous radiological diagnosis of fibrous dysplasia for years. An aggressive variant of systemic mastocytosis was diagnosed on bone marrow examination. Systemic mastocytosis was confirmed by splenectomy, liver biopsy and finally autopsy. The present case is unique because of the ovarian location of the germ cell tumour. We suggest our observation could be related to the broad group of haematological malignancies associated with germ cell tumours. PMID- 9205865 TI - Giant (bizarre) cell variant of renal carcinoma. PMID- 9205866 TI - Granulomatous visceral neuropathy of the colon with non-small cell lung carcinoma. PMID- 9205867 TI - Kimura's disease of the epiglottis. PMID- 9205868 TI - Spindle and epithelioid haemangioendothelioma of the inguinal lymph nodes. PMID- 9205870 TI - Detectability of retinoblastoma (Rb) gene product and p53 antigen expression decreases in stored paraffin sections. PMID- 9205869 TI - Langerhans cell histiocytosis of lymph nodes: draining a papillary carcinoma of the thyroid. PMID- 9205871 TI - Estimating mitotic activity in tumours. PMID- 9205872 TI - Endocrine cell carcinoma (carcinoid tumour) of the gallbladder producing pancreatic polypeptide and somatostatin. PMID- 9205873 TI - Vascular surgery for impotence: a review. AB - Progress in treatment of impotence in the past two decades has resulted in impressive advances. While most men respond to medical therapy including prostaglandin E1 injection or the more recent use of urethral alprostadil, 6-7% of men fail to respond to these treatments or vacuum devices. This review considers current and past results of vascular surgery in this group of men. Guidelines for case selection for vascular interventions as well as reporting criteria are suggested. Vascular surgery as a logical first step in selected patients may offer an advantage in men failing conservative therapy and for those not desiring prosthetic implantation. PMID- 9205874 TI - Integrative erectile biology: the role of signal transduction and cell-to-cell communication in coordinating corporal smooth muscle tone and penile erection. AB - The contractility of corporal smooth muscle plays a critical role in the entire erectile process in man. Moreover, in the absence of severe vascular disease, or congenital or other structural abnormalities/malformations, relaxation of the corporal smooth muscle is both necessary and sufficient to elicit a sustained erection. As such, understanding the initiation, maintenance and modulation of corporal smooth muscle tone is an absolute prerequisite to the improved understanding, diagnosis and treatment of erectile dysfunction. Despite this fact, identification of both the precise mechanistic basis by which endogenous and exogenous vasomodulators exert their effects on individual corporal smooth muscle cells, and moreover, the process by which these signals are spread among the diverse array of parenchymal cells in the paired corpora, remains somewhat of a physiological enigma. Therefore, the goal of this report is two-fold: first, to review current knowledge of the regulation of corporal smooth muscle tone at the cellular and molecular level; and second, to outline a cogent explanation for the rapid and syncytial integration of the effects of diverse stimuli among corporal smooth muscle cells in the human penis. PMID- 9205875 TI - Vascular control mechanisms in penile erection: phylogeny and the inevitability of multiple and overlapping systems. AB - A co-ordinated series of vascular events underlie the generation of a penile erection. The control and regulation of this simple event is, in fact, a complex of interactions occurring at multiple levels. Many of these individual pathways and responses have been studied extensively. The understanding of the necessity of the integration between the individual pathways into a complex of series and parallel coupled mechanisms provides a rationale for the development of a framework of multiple and overlapping systems. This paper sets out some of the principles of integrated and balanced control of vasodilation and vasoconstriction in the penis. In addition, the role of growth induction and regression and the importance of time as a factor in studying penile structure and function is discussed. PMID- 9205876 TI - Radiologic evaluation of penile arterial anatomy in arteriogenic impotence. AB - Functional and anatomic evaluation of penile arterial blood flow is essential in the work up of erectile dysfunction. Duplex ultrasonography is an ideal screening modality with cavernosal mean peak systolic blood flow velocity being the most sensitive predictor of arterial disease. Arterial variability of the penis may affect sonographic evaluation leading to frequent misinterpretation and therefore pudendal arteriography remains the current gold standard for penile arterial evaluation. Appreciation of the type and frequency of anatomic variants and potential collateral routes is important in interpreting penile arteriograms and in evaluating the hemodynamic significance of suspected arterial disease. PMID- 9205877 TI - Use of radioactive tracers in the evaluation of penile hemodynamics: history, methodology and measurements. AB - Radionuclide tracer techniques are intimately associated with some of the early ground-breaking investigations in erectile dysfunction and have evolved along with the field. At the present time, the various investigations can be grouped into four categories: labeled blood-pool; tracer washout; tracer washin and combined blood-pool/tracer and tracer washout examinations. Blood pool studies are most useful in assessing the integrity of arterial inflow, but may also be used to generate indices of venous leak. A non-imaging version of the blood-pool test may represent a simple and cost-effective alternative. Washout of intracavernosal xenon during erection seems the most rigorous method of testing venous integrity. Washout using 99mTc-labeled substances may emerge as a convenient alternative to the more technically difficult xenon examinations. Dual isotope blood pool and washout examinations, though complicated, represent an ideal method of analyzing penile hemodynamics, with potential to contribute significantly to the understanding of penile physiology. Development of improved pharmacologic stimuli and augmentation of testing protocols by intracavernosal pressure monitoring may further improve the utility of quantitative and physiologic nuclear medicine examinations in erectile dysfunction. PMID- 9205878 TI - Oxidation effects in fatigue: unphysiological responses to "depolarization" in skinned muscle fibers. PMID- 9205879 TI - Berlin symposium on radiotherapy of ocular diseases. An encouraging document of steady progress by means of highly developed interdisciplinary treatment approaches. PMID- 9205880 TI - High-resolution MR imaging of the eye and orbit. PMID- 9205881 TI - A special surface coil for high-resolution ocular MRI. PMID- 9205882 TI - Dosimetry and design of radioactive eye plaques. PMID- 9205883 TI - Stereotactic precision radiotherapy in the treatment of intraocular malignancies with a micro-multileaf collimator. PMID- 9205884 TI - Stereotactic irradiation of uveal melanoma with the Leksell gamma unit. PMID- 9205885 TI - Eye preservation brachytherapy for orbital and adjacent tumors: preliminary results. PMID- 9205887 TI - Whole-eye versus lens-sparing megavoltage therapy for retinoblastoma. PMID- 9205886 TI - Precision megavoltage external beam radiation therapy for retinoblastoma. PMID- 9205888 TI - Retinoblastoma after chemoreduction and irradiation: preliminary results. PMID- 9205889 TI - Radiotherapy for retinoblastoma. Treatment strategies. PMID- 9205890 TI - Metastatic disease, eye retention and visual function in conservative treatment of uveal melanoma. PMID- 9205891 TI - Results of proton radiotherapy for uveal melanomas. PMID- 9205893 TI - Clinical and technical requirements for proton treatment planning of ocular diseases. The SERAG (South Europe Radiotherapy Group). PMID- 9205892 TI - Adjunctive plaque radiotherapy after local resection of uveal melanoma. PMID- 9205895 TI - Treatment of advanced conjunctival melanoma by external beam irradiation. PMID- 9205894 TI - Ruthenium-106 eye plaque brachytherapy in the conservative treatment of uveal melanoma: evaluation of 175 patients treated with 150 Gy from 1981-1989. PMID- 9205896 TI - Radiotherapy for choroidal metastases: interim analysis of a prospective study of the ARO(ARO 95-08). PMID- 9205897 TI - Bilateral radiotherapy in cases of one-sided choroidal metastases. PMID- 9205898 TI - Strategies for diagnosis and management of infiltrative orbital lesion. PMID- 9205899 TI - Radiation therapy of orbital lymphoid tumors: introduction of lens-sparing techniques in Hungary. PMID- 9205900 TI - Radiotherapy of pseudotumor orbitae. PMID- 9205901 TI - Radiotherapy of primary orbital lymphomas. PMID- 9205902 TI - Clinical importance of radiotherapy in the treatment of Graves' disease. PMID- 9205904 TI - Radiotherapy for severe, progressive thyroid-associated ophthalmopathy: long-term results with comparison of scoring systems. PMID- 9205903 TI - Results and prognostic factors in retrobulbar radiotherapy combined with systemic corticosteroids for endocrine orbitopathy (Graves' disease). PMID- 9205905 TI - Teletherapy in the management of patients with age-related macular degeneration complicated by subfoveal neovascularisation: an overview. PMID- 9205907 TI - External radiotherapy in macular degeneration: technique and preliminary subjective response. PMID- 9205908 TI - Radiotherapy of macular degeneration. Experience with two fractionation schemes. PMID- 9205906 TI - Stereotactic radiation therapy for subfoveal choroidal neovascularization secondary to age-related macular degeneration. PMID- 9205909 TI - Postoperative pterygium prevention by radiotherapy with strontium-90 beta-rays. PMID- 9205910 TI - Radiation therapy for choroidal and retinal hemangiomas. PMID- 9205911 TI - Acute and late side effects of radiotherapy for ocular disease: an overview. PMID- 9205912 TI - Side effects of photon and proton radiotherapy for ocular melanoma. PMID- 9205913 TI - Bone morphogenetic proteins: background and implications for oral reconstruction. A review. AB - For over 30 years now, research has been carried out to isolate and purify bone morphogenetic protein (BMP), a substance which has been shown to induce heterotopic bone formation in various animal species. Recent advances in the fields of developmental biology, molecular biology, genetics and wound healing, have shown that the BMPs are not only responsible for postfetal bone induction (including normal bone remodeling, healing and repair), but are also critical during embryogenesis, not only in regards to the skeletal system, but quite possibly in the morphogenesis and pattern formation of other tissues and organs as well. Therefore, BMPs have the potential as a therapeutic utility in orthopedic and dento-alveolar reconstruction. PMID- 9205914 TI - Retrospective analysis of factors related to clinical outcome of guided tissue regeneration procedures in intrabony defects. AB - The purpose of this retrospective study was to determine factors affecting clinical outcome of guided tissue regeneration (GTR) in the treatment of intrabony periodontal defects. 38 patients each contributing 1 isolated intrabony defect treated with GTR were included in this analysis. Patient and defect characteristics, and defect-specific recordings relative to clinical outcome 6 months postsurgery were assessed. GTR treatment resulted in clinically and statistically significant improved probing depths (PD), clinical attachment levels (CAL), and probing bone levels (PBL). Presurgery PD and PBL were of predictive value for CAL gain and PBL gain, respectively. CAL and PBL gain did not correlate to defect depth or configuration. Cigarette smoking exhibited a highly significant negative correlation to parameters of clinical outcome. PMID- 9205915 TI - Guided tissue regeneration in conjunction with hydroxyapatite-collagen grafts for intrabony defects. A clinical and radiological evaluation. AB - This clinical and radiological study evaluated the healing of 3 + 2 + 1 wall combined intrabony defects treated using the guided tissue regeneration technique (GTR) with and without hydroxyapatite-collagen alloplastic graft materials (HAC), in comparison to that of HAC alone and conventional flap surgery (CF). 40 interproximal defects with probing depth > 6 mm were treated in 18 adult periodontitis patients of ages 35-60 years. After non-surgical therapy, the defects were randomly grouped into 4 groups of 10 defects each. These groups were designated: (1) expanded polytetrafluoroethylene membrane (e-PTFE), (2) e-PTFE + HAC, (3) HAC alone and (4) CF. At 6 months, the following changes in parameters were recorded. Mean PPD reduction for each group was 5.83, 5.85, 3.80 and 3.17 mm respectively. PPD reduced very significantly in all groups (p < 0.01), the highest and lowest reductions in PPD being for the e-PTFE + HAC and CF group respectively. Comparison between the 4 groups showed higher PPD reduction in both membrane groups than in either of the non membrane groups (p < 0.05) with the difference between the e-PTFE and CF groups being very highly significant (p < 0.001). Mean attachment gain for the 4 groups was 3.70, 3.80, 2.60 and 2.1 mm, respectively. Similarly attachment gain for all groups was very significant (p < 0.01) and the highest and lowest attachment gains were for the e-PTFE + HAC and CF group respectively. Both membrane groups showed significantly more attachment gain than the CF group (p < 0.05). Change in probing bone level (BL) for the 4 groups was 1.60, 1.90, 1.0 and 0.65 mm respectively. Again the highest changes in BL were recorded for the e-PTFE + HAC group. Significant differences were found between both membrane groups and the CF group (p < 0.05). Radiological evaluation using standardized radiographs and millimeter grids showed change in radiographic bone level at the deepest point of the defect on the radiograph to be 1.50, 1.55, 0.85 and 0.60 mm, respectively and this was significantly higher in both membrane groups than in the CF group (p < 0.05). This study therefore found e-PTFE membranes both alone and when combined with HAC to lead to more attachment gain and bone fill than did HAC alone or CF. It found HAC combined with e-PTFE to perform better although not significantly better than e-PTFE alone. PMID- 9205916 TI - Multifocal eosinophilic granuloma with sequential periodontitis-like lesions. AB - Eosinophilic granuloma (EG) is the localized and mildest form of the triad commonly known as Langerhans cell histiocytosis. This report describes a case manifesting itself as a periodontal problem with the localized osseous lesions in jawbones which was first diagnosed as early-onset periodontitis. Later on, the diagnosis of EG was established, relying on histopathological and immunohistochemical evaluations. Immunohistochemical findings confirm that a minor component of cell aggregates is phenotypically related to Langerhans cells among the sheet-like accumulations of histiocytes and eosinophils. The aetiology of the disease remains largely unknown. Although surgical curettage of lesions is usually effective in treatment of EG of bone, corticosteroids might be used as an adjunctive. This multifocal case of EG stresses the importance of clinical follow up examinations, since the sequential lesions appear with irregular intervals, and this may cause diagnostic problems as well as a delay in starting the treatment regimen. PMID- 9205918 TI - An investigation of bioadhesion for periodontal and oral mucosal drug delivery. AB - Gel delivery vehicles have ideal placement characteristics for periodontal and oral mucosal drug delivery. However, the retention of the vehicle at the site may be of short duration thereby limiting its therapeutic effect. Bioadhesion has received little attention as a means of enhancing vehicle retention in the periodontal pocket and this study aimed to investigate the possible role of this phenomenon to aid oral drug delivery. Chitosan, xanthan gum and poly (ethylene oxide) were selected as potential vehicles from previous in vitro studies, since all 3 had shown good bioadhesive properties. Retention in the periodontal pocket was assessed by means of an insoluble fluorescein marker in 8 patients, and to the oral mucosa by the retention of a small plastic film in 12 subjects. The results showed that fluorescein release from the periodontal pocket was significantly longer for chitosan than for other gels or a water control. In contrast, xanthan gum gave the most prolonged adhesion time on the oral mucosa (153.5 min) followed by poly (ethylene oxide) (89.3 min) and chitosan (42.6 min), and these times were all significantly different from each other (p < 0.05). The results from this study would tend to suggest that the bioadhesive properties of an aqueous gel may be directly related to its retention both in the periodontal pocket and on the oral mucosa. However, other important factors for mucosal adhesion include the patient acceptability of the formulation and the choice of application site. PMID- 9205917 TI - Possible autosomal-dominant inheritance of prepubertal periodontitis in an extended kindred. AB - This study presents the clinical findings and the distribution of prepubertal periodontitis in an extended family with high prevalence of this entity. The expression of surface markers and adhesion molecules on peripheral lymphocytes were also studied. Approximately 50% of the children in this family suffered from prepubertal periodontitis. All the affected children were otherwise healthy. 2 identical twins were similarly, but not identically, affected. Detailed laboratory tests and analysis of lymphocyte surface marker expression, including CD18, were all within the normal levels. Both localized and generalized forms of prepubertal periodontitis were found. The high prevalence of prepubertal periodontitis in the 2 branches of this family, and the fact that identical twins were similarly affected, suggest a strong genetic predisposition for prepubertal periodontitis. The family pedigree is consistent with an autosomal-dominant mode of transmission. The coexistence of localized and generalized forms of the disease in sibs suggests the same genetic etiology for both entities with variability in disease expression. This variability in disease expression is further supported by the fact that 2 identical twins were not identically affected. PMID- 9205919 TI - Antimicrobial susceptibility tests on anaerobic oral mixed cultures in periodontal diseases. AB - The ecosystem of the dental plaque in periodontal diseases is very complex: the study of such micro-organisms, which are mostly strict anaerobes, requires the use of specific techniques under conditions of strict anaerobiosis. The aim of the present study was to design a rapid method to evaluate the activity of antimicrobials on mixed bacterial plaque of subjects with periodontal diseases. The study was carried out using a computerised instrument generally used for simultaneous diagnostic tests with aerobic bacteria. Operative and methodological modifications were made to obtain conditions of strict anaerobiosis and the balanced growth of all the microbial forms present in the mixed cultures of the plaque. Penicillins and cephalosporins were active on all the samples, whereas colistin, gentamicin, kanamycin and nalidixic acid showed no activity. Clindamycin, tetracycline, erythromycin and penicillin G were effective only against some samples. The activity of the antimicrobials towards isolated strains was analogous to that towards the corresponding mixed culture. PMID- 9205920 TI - Particulate bioglass as a grafting material in the treatment of periodontal intrabony defects. AB - The present clinical trial was designed to evaluate the effects of a bioactive glass, Perioglas, in the treatment of periodontal intrabony defects. 20 patients, 23-55 years of age (44 sites), with intrabony defects completed the 1-year study. Teeth with furcation involvement were excluded. After completion of initial therapy, defects were randomly assigned to either a test or control procedure. Following flap reflection, root planing and removal of chronic inflammatory tissue in both groups, the test defects were restored with the bioactive glass particulate material. Mucoperiosteal flaps were replaced, sutured and a periodontal dressing was used. All the patients received postoperative antibiotics and analgesics and were seen at 1 week for suture removal. Follow-up was then carried out weekly and at 3 months, 6 months, 9 months and 1 year post surgery. Plaque score, bleeding score, probing pocket depth (PPD), probing attachment level (PAL) and gingival recession were recorded at baseline, 3 months and 1 year. Standardised radiographs for computer-assisted densitometric image analysis (CADIA) were taken at baseline, immediately post-operatively and at 1 year. The CADIA data showed a significant increase (F-ratio: 15.67, p < 0.001) in radiographic density and volume between the defects treated with the Perioglas when compared to those treated with surgical debridement only. PPD and PAL showed significant improvements in both experimental and control sites, with a greater trend to improvement in the experimental sites. It was concluded that this bioactive glass is effective as an adjunct to conventional surgery in the treatment of intrabony defects. PMID- 9205921 TI - A clinical evaluation of a novel toothbrush design. AB - A new toothbrush design, the Snakebrush, was clinically evaluated along with the Flexible and Precision brushes. In this parallel, random-allocation investigation, the plaque removing qualities of the 3 brushes were evaluated over a period of 30 days. 60 patients of good dental health between the ages of 20 and 30 years formed the basis of the clinical trial. Mean plaque area scores were evaluated and recorded after a 1-min brush at baseline, day 15 and day 30. Gingivitis indices and bleeding on probing indices were also recorded at the same time intervals. Both uni- and multivariate analyses of our data suggested that the Snakebrush removed significantly more plaque than the 2 controls. For both lingual and buccal surfaces, subjects who had used the Snakebrush showed a significant decline in bleeding on probing and qinqivitis indices. PMID- 9205922 TI - Investigations into the cellular contribution to host tissue proteases and inhibitors in gingival crevicular fluid. AB - Gingival crevicular fluid (GCF) was collected from chronic periodontitis patients using plastic micropipettes and coverslip smears stained with antibodies for leukocyte markers and Toluidine Blue for mast cells. The smears consisted of 70 80% granulocytes, 10-20% monocytes/macrophages, 5% mast cells and 5% T lymphocytes; no B lymphocytes were found. Proteases and inhibitors in GCF cells were investigated by enzyme cytochemistry using 2-methoxy-4-naphthylamine-linked peptide substrates and simultaneous coupling to Fast Blue B and immunocytochemistry using biotinylated secondary antibodies and an alkaline phosphatase/new fuchsin detecting system. Elastase was detected in granulocytes, cathepsin B in macrophages, dipeptidyl peptidases II and IV in a small proportion of macrophages, dipeptidyl peptidase IV in a few T lymphocytes, tryptase in mast cells and alpha-1-proteinase inhibitor and alpha-2-macroglobulin in some macrophages. GCF was also collected on filter paper strips and eluted into buffer for biochemical enzyme assays. Lysis of cells by addition of detergent to the elution buffer increased activities to 140-240% of control values. Removal of cells by centrifugation reduced measured activities to 1-30% of original figures; this effect was less if samples were pre-treated with detergent. Proteases from inflammatory cells therefore appear to make up most of the measured enzyme activity in GCF, and this association may explain recent correlations with periodontal disease progression. PMID- 9205923 TI - The bactericidal effects of dental ultrasound on Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. An in vitro investigation. AB - This study investigated the possible bactericidal acoustic effects of the dental ultrasonic scaler. Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis suspensions, were subjected to the vibrations of a Cavitron P1 insert for 2.5 and 5.0 min in an acoustically-simulated pocket model and the survivors enumerated. The extent of any cavitation occurring within the pocket model to which the statistically significant bactericidal activity observed might be attributed, was determined by 'sonoluminescence', which was then investigated by photomultiplication techniques. However, these failed to detect any sonoluminescence within the pocket space and, moreover, the necessary deflection of the water coolant away from the insert tip, to avoid flooding of the experimental pocket, proved to result in temperatures of 47.6 degrees C and 52.3 degrees C at the respective time intervals, and thereby constituted an alternative possible bactericidal mechanism. Examination of the effects of such temperature changes on the target bacteria then revealed statistically significant differences in the viable counts of both microorganisms after 5.0-min periods, and as such were comparable to those previously detected in relation to the pocket model. Whilst it must be presumed that the bacteriolytic effect observed in the main investigation was due to the incidental temperature changes, in the absence of acoustic cavitation the influence of any associated acoustic microstreaming cannot be discounted. Further investigations to assess the bactericidal potential of acoustic phenomena using a modified experimental to exclude any hyperthermic effects are therefore necessary. PMID- 9205924 TI - A clinical evaluation of a bioresorbable barrier with and without decalcified freeze-dried bone allograft in the treatment of molar furcations. AB - This study evaluated a bioresorbable barrier with and without decalcified freeze dried bone allograft (DFDBA) in the treatment of human molar furcations. 14 subjects with paired class II mandibular molar furcation defects participated in the study (8 male and 6 female). The class-II furcation defects were randomly treated with either the resorbable barrier alone or resorbable barrier in combination with decalcified freeze-dried bone allograft (DFDBA). Gingival recession, probing depth, clinical attachment, and bone fill were measured 6 months post-treatment measurements were repeated and each site was surgically re entered. When the resorbable barrier alone was compared to resorbable barrier in combination with DFDBA, probing depth reduction was significantly (p < 0.01) in favor of the combination therapy. Vertical bone gain was significant with the combination treatment demonstrating more bone fill (p < 0.02). Smoking was also a variable examined in this study. When compared to smokers, non-smokers for both treatment groups revealed greater probing depth reduction, vertical bone gain, and horizontal bone gain. Within the non-smoking group, probing depth reduction was also significantly higher for the resorbable barrier and DFDBA group than the resorbable alone group (p < 0.02). These results illustrate that the probing depth reduction is better in the non-smoker and the best in the non-smoker with the combination therapy of resorbable barrier and DFDBA than with resorbable barrier alone. PMID- 9205925 TI - Homosexuality and totalitarianism. AB - Since the dissolution of Communism left the Right with no unifying enemy, some conservatives may be turning to the gay world as the new "evil empire." This article explores the ideology that underpins the recent upsurge of gay-baiting. Its central thesis is that in identifying the homosexual as the demonic "other" the cultural Right is offering us another form of ersatz Christianity masquerading as tradition but actually infected with the virus of modernity. The paper classifies homophobia as an essentially secular phenomenon, reminiscent of other ideologies which have arisen as a substitute for religion. PMID- 9205926 TI - Jewish responses to AIDS. AB - Theoretically, Judaism treats AIDS, an illness, with compassion. Yet because of its link to homosexuality, AIDS and its stigma are often concealed. Neither homosexuality nor AIDS can be fully concealed, even by those who most denounce them, nor can they be completely open, even by those who most fully accept them. The implications of rejection and acceptance (a continuum) are discussed. In spite of the biblical rejection of homosexuality, The Gay Synagogue offers the most meaningful support to those who suffer from AIDS and to their loved ones. Paradoxically, the most successful programs are those adhering to the formal religious practices which provide the basic structure to every Jewish synagogue. PMID- 9205927 TI - Lesbian uses of and satisfaction with mental health services: results from Boston Lesbian Health Project. AB - In response to the dearth of specific information about lesbians' health status and practices in the health literature, a national study utilizing a self administered questionnaire was conducted in 1987 by four associates of the Fenway Community Health Center in Boston to access data in these areas. The questionnaire solicited information about demographics, health practices, stress in personal and work lives, mental and physical health problems, sexual practices, family history of health related problems, and genetic attributes hypothesized to be related to "gayness." Questionnaires from 1,633 lesbian women provided the database for the study. This paper discusses the portion of the survey that dealt with mental health services and life experiences. Past studies that investigated mental health needs of lesbians focused on the quality of treatment by mental health providers, rates of suicide attempts, and alcoholism. This paper compares these past findings with the responses of the lesbians in this national, community-based study. Findings indicate that although a significant number of the lesbian women in this sample had been in therapy, they sought out therapy as a coping strategy to deal with similar issues as other women, i.e., depression and relationships. Suicide attempts decreased considerably after adolescence and "coming out." Rates of alcohol use and abuse, although difficult to compare with other studies, were higher than other women but similar to other studies investigating a community sample of lesbians. Even with a high family history of alcoholism, less than 5 percent reported having sought out therapy to deal with any issues of alcohol or drug use. PMID- 9205928 TI - Rural gay men in northern New England: life experiences and coping styles. AB - This study describes thematically the life experiences of 20 gay men in the rural setting of northern New England and examines what coping skills they have evolved. A qualitative study was undertaken, so that the researchers could learn of rural gay men in their own words, particularly in terms of how they understood their life experiences. This material was analyzed and 9 common themes were discovered. In descending order of frequency of occurrence in subjects' narratives, the themes are: early awareness of difference, internalized homophobia, positive aspects of rural living, negative aspects of rural living, positive family of choice, compulsory heterosexuality, isolation, current life partner, and family censorship. PMID- 9205929 TI - The formation of gay and lesbian identity and community in the Connecticut River Valley of western Massachusetts, 1900-1970. AB - The formation of lesbian and gay identity and community in the Connecticut River Valley of western Massachusetts was greatly shaped by social changes and trends in gender ideology which originated outside the region. Safe and supportive space for the exploration of homosexual identity was limited and limiting, as gay and lesbian residents turned to homoerotic communities away from the area to try to come to terms with and act out their same-sex desires. Gay men and lesbians in the Valley began to generate self-affirming and politically oriented institutions, however, within the context of the radical political culture of the 1960s. PMID- 9205930 TI - Frederick Henry Horatio Akbar Mahomed: 'a brave and ambitious explorer'. PMID- 9205932 TI - Doctors' attitudes towards the detection and treatment of hypertension in older people. AB - Within the former northern region a questionnaire was sent to a one in two random sample of general practitioners and to consultant physicians involved in the treatment of older hypertensive patients, to determine current attitudes of doctors to the management of hypertension in older people. A total of 407 general practitioners and 125 consultant physicians (including 38 consultant geriatricians) completed the questionnaire; response rates of 58% and 73% respectively. The median (range) threshold level for intervention of an otherwise well 75-year-old patient was 180 (140-240)/100 (90-120) mm Hg for general practitioners as against current recommendations of > or = 160/90 mm Hg. Geriatricians adopted significantly lower thresholds than both general practitioners and other specialists. Target blood pressure (BP) and drug therapy were in line with British Hypertension Society recommendations. The majority of general practitioners stated that they would initiate treatment for hypertension later, be less aggressive in their treatment and accept higher levels of BP than with younger patients. Approximately half perceived compliance and lifestyle changes as problematic in older people, while approximately a third were concerned about the problems created by drug treatment and uncertainty about the practicality of treating all older hypertensive patients. A quarter expressed reluctance to initiate hypertensive treatment because of side effects and to treat isolated systolic hypertension. These stated attitudes amongst general practitioners may explain the continuing conservatism in the management of hypertension. This study found considerable variation in diagnostic threshold between doctors, which suggests that current management of hypertension in older people is likely to vary significantly. PMID- 9205931 TI - Therapy in old patients with isolated systolic hypertension: fourth progress report on the Syst-Eur trial. AB - The Syst-Eur trial is a multicentre, randomized, double-blind, placebo controlled therapeutical trial in patients at least 60 years old and with isolated systolic hypertension. Its scope is to investigate the effects of modern antihypertensive drug treatment on morbidity and mortality and to assess possible adverse effects of the drugs used. Patients were recruited in 22 countries in western and eastern Europe and Israel. At three run-in visits 1 month apart their sitting systolic blood pressure (SBP) on single-blind placebo treatment averaged 180-219 mm Hg with diastolic blood pressure (DBP) lower than 95 mm Hg. After stratification for sex and the presence of cardiovascular complications, the patients were randomized either to active treatment or placebo. Active treatment consisted of nitrendipine (10-40 mg/day) with the possible addition of enalapril (5-20 mg/day) and/or hydrochlorothiazide (12.5-25 mg/day), titrated or combined to reduce the sitting SBP by at least 20 mm Hg to below 150 mm Hg. Matching placebos were employed similarly. The present progress report is based on the data received at the Coordinating Office before 1 March 1996. At that time 3433 subjects had been randomized. A total of 2015 patients had been followed for at least 1 year on double-blind treatment and 1298 patients for at least 2 years. At baseline BP was similar in both treatment groups and averaged 174/86 mm Hg. According to a per protocol analysis at 1 year, BP fell (P < 0.001) on average by 22.6 +/- 15.7/6.0 +/- 8.0 mm Hg in the active treatment group and by 12.2 +/- 15.9/1.7 +/- 7.3 mm Hg in the placebo group. At 2 years BP was 10.2/5.7 mm Hg lower (P < 0.001) on active treatment than on placebo. At 1 year the percentage of patients who had reached goal BP was 19.9% in the placebo group and 41.4% in the active treatment group. At 2 years these percentages were 20.9 and 43.2 respectively. PMID- 9205933 TI - Hypertension awareness and control in an inner-city African-American sample. AB - African-Americans in the US are at high risk for hypertension-related morbidity and mortality. The majority of African-Americans live in central city areas, and lower socioeconomic status and health care utilization patterns have been hypothesized to contribute to higher blood pressure (BP) levels and poorer control of treated hypertension in this group. In order to plan an intervention to improve hypertension care for inner city African-Americans in Houston, Texas, we conducted a baseline survey of residents in 12 low-income ZIP code areas with a > 70% African-American population to determine the level of hypertension awareness, treatment and control, and associated sociodemographic, health care utilization, and medication compliance variables. Subjects were recruited to attend a BP measurement and assessment of knowledge, attitudes and behaviors through random digit phone dialing in the target ZIP code areas. Of the 962 subjects examined, 433 (45%) were hypertensive (systolic BP > or = 140 mm Hg or diastolic pressure > or = 90 mm Hg or taking antihypertensive medication). Among all hypertensives, 73% were aware, 64% were on treatment, and 28% were controlled to 140/90 mm Hg. Of hypertensives on treatment, 43% were controlled to 140/90 mm Hg, but 72% were controlled using the criterion of 160/95 mm Hg, and 75% were controlled using a diastolic pressure < 90 mm Hg only. These results are similar to those reported for African-Americans in the most recent US national health survey. Males were less likely to be aware, receiving treatment and controlled than were females. Although lack of awareness was associated with less frequent BP measurement, 77% of those unaware reported a measurement within the past 2 years. The majority of aware hypertensives reported frequent physician contact and high compliance with medication. We conclude that intervention to improve hypertension control in this population should focus on ensuring that health providers diagnose BP and establish treatment goals based on the current standard of 140/90 mm Hg. PMID- 9205934 TI - Prevalence of positive Osler's manoeuver in 3387 persons screened for the Systolic Hypertension in the Elderly Program (SHEP) AB - The purpose of this study was to examine the prevalence of a positive Osler's manoeuver (OM) among persons screened for the Systolic Hypertension in the Elderly Program (SHEP). Information obtained from all individuals included age, gender, and race; history of antihypertensive, anticoagulant, insulin, or cardiac pacemaker use; and history of heart attack, coronary bypass surgery, or stroke. Among the persons aged 60 and over that were screened for eligibility at the Louisville SHEP Center, OM was performed on 3387 subjects. Of these, 7.2% (243 of 3387) were determined to be Osler maneuver positive (O+). O+ was more prevalent among males than among females (P = 0.025). A higher prevalence of O+ was associated with both higher age (P < 0.001) and higher blood pressure (P < 0.001). There were significantly more Osler positives among those who responded positively to 'Have you had a stroke?' (P = 0.007) and 'Are you taking anticoagulants' (P = 0.044) than among those who responded negatively to these questions. O+ was also less prevalent among those that were normotensive at the time of the screening (P = 0.046). The results of this study, when taken with those of previous studies of OM, support the cautious use of OM as a screening tool for pseudohypertension in the elderly population and as an adjunct in determining the cardiovascular risk profile of individual patients. However, further study of OM is required before it can be recommended for routine use in the assessment of hypertension among the elderly. PMID- 9205935 TI - Antihypertensive therapy and orthostatic responses in elderly hospital in patients. AB - The study aim was to determine the association between use of antihypertensive drugs and orthostatic hypotension on prolonged standing in an elderly in-patient population. Hospital in-patients aged > 60 years had manually and automatically determined blood pressure (BP) measurements recorded in the supine position. On standing a total of nine measurements were taken over 10 min, six measurements were taken using a mercury sphygmomanometer and three by an automatic monitor. Seventy-four patients of mean age 73 +/- 7 years were studied; 52 (70%) were taking > or = 1 antihypertensive drug and 22 (30%) none. On standing, manually determined systolic BP (SBP) fell to a similar extent in the group of patients taking antihypertensive therapy compared to those not taking such treatment (at 9 min standing: -6 +/- 16 vs -10 +/- 15 mm Hg, respectively) and the frequency of orthostatic hypotension (SBP fall > or = 20 mm Hg) was similar in both groups [at 9 min: 9 (17%) vs 5 (23%)]. Automatically determined measurements also revealed similar orthostatic SBP responses in both treated and non-treated groups (at 8 min: -3 +/- 18 vs -6 +/- 13 mm Hg, respectively) and a similar frequency of orthostatic hypotension. No significant change in standing compared to supine diastolic BP (DBP) measured manually or automatically was seen in either group. Even in the subgroup of patients taking > or = 2 antihypertensive drugs the orthostatichypotension. BP response and the frequency of orthostatic hypotension was similar to that in the non-treated group. In conclusion no association was found between use of antihypertensive therapy and orthostatic hypotension in an elderly in-patient population. PMID- 9205936 TI - Fourier analysis of circadian blood pressure profile in secondary hypertension. AB - Different types of statistical methods have been used for circadian blood pressure (BP) rhythm analysis in secondary forms of hypertension. In the present study, we used the two-step statistical approach by Fourier analysis with four harmonics for the parametrization of the diurnal BP pattern in secondary hypertension. In 43 essential hypertensives (EH), eight patients with aldosterone producing adenoma (APA), 25 with idiopathic hyperaldosteronism (IHA), four with glucocorticoid remediable hyperaldosteronism (GRH) and seven with renovascular hypertension (RVH), 24-h ambulatory BP was measured. The diurnal BP and heart rate (HR) rhythm was present in more than 70% of patients with secondary hypertension, without significant differences with EH and despite the attenuation in the degree of the nocturnal BP fall. In conclusion, the statement that secondary hypertension is characterized by an abnormal diurnal rhythm of BP is a gross over-simplification. Our findings suggest that the two-step method with four harmonics Fourier analysis may represent a useful method and a more complete statistical approach to providing circadian parametrization of the 24-h profile in secondary hypertension. PMID- 9205937 TI - Serum angiotensin-converting enzyme activity correlates positively with plasma angiotensin II: a population-based study of ambulatory blood pressure and the renin-angiotensin system. AB - A population-based study was performed in order to study the interrelationships of the circulating components of the renin-angiotensin system during basal conditions and their relations to blood pressure (BP), age and gender. One hundred and four women and 95 men, 16-70 years old, evenly age distributed and randomly selected from the population of Linkoping, Sweden, participated. Venous blood was drawn at 08.00 hours and ambulatory BP recording was then performed. Serum angiotensin-converting enzyme (ACE) activity correlated with plasma angiotensin II (r = 0.20, P = 0.004), but when calculated separately according to gender, the correlation remained significant only in men (r = 0.33, P = 0.001). Plasma renin activity (PRA) correlated negatively with age (r = -0.30, P < 0.0001), but immunoreactive active renin (IRR) and angiotensin II did not. PRA and IRR correlated negatively with BP in women but correlations disappeared after age adjustment. The 23 women on oestrogen medication did not differ from the remaining 81 with respect to age (P = 0.5), IRR (P = 0.96) or angiotensin II (P = 0.4) levels, but PRA was higher (2.2 +/- 1.4 ng Ang l/ml/h and 1.5 +/- 0.9 ng Ang l/ml/h, respectively, P = 0.004). PRA (r = 0.38, P < 0.0001) and IRR (r = 0.49, P < 0.0001) correlated positively with the levels of angiotensin II. In conclusion the fact that PRA, but not IRR, declined with age and was higher among oestrogen treated women, although angiotensin II was unaffected suggests IRR to be a more robust marker of angiotensin II levels than is PRA in a population-based setting. ACE correlates positively with angiotensin II in men. PMID- 9205938 TI - Losartan mediated improvement in insulin action is mainly due to an increase in non-oxidative glucose metabolism and blood flow in insulin-resistant hypertensive patients. AB - We investigated the possible role of losartan on insulin-mediated glucose uptake, substrate oxidation and blood flow in insulin-resistant hypertensive patients. Sixteen newly diagnosed patients with mild-to-moderate hypertension were studied. The study design was a single-blind, randomised, placebo-controlled trial. After a 1 week run-in period, each patient was randomly assigned to placebo (n = 7) and losartan (n = 9). Both treatment periods lasted 4 weeks. At baseline, and at the end of the placebo and losartan treatment periods, euglycaemic hyperinsulinaemic glucose clamp and indirect calorimetry were performed. Before and along each glucose clamp, blood flow was also determined in the femoral artery by image directed duplex ultrasonography combining B-mode imaging and pulse Doppler beams. Losartan vs placebo lowered systolic blood pressure by 163 +/- 3.5 and 147 +/- 4.1 mm Hg (P < 0.001), and diastolic blood pressure by 95 +/- 3.2 and 85 +/- 3.2 mm Hg (P < 0.001). Losartan enhanced glucose metabolic clearance rate by 5.1 +/- 0.3 and 6.3 +/- 0.4 mg/kg x min (P < 0.05), and whole body glucose disposal (WBGD) by 29.2 +/- 0.5 and 38.1 +/- 0.4 mumol/kg free fatty mass (FFM) x min (P < 0.01) but did not affect heart rate. Insulin-mediated change in blood flow was greater after losartan than placebo administration (111 +/- 4 vs 84 +/- 3%, P < 0.01). Per cent change in insulin-mediated stimulation of blood flow and WBGD were also correlated (r = 0.76, P < 0.01). Analysis of substrate oxidation revealed that losartan administration improved insulin action and non-oxidative glucose metabolism (NOGM) (30.8 +/- 2.2 vs 22.8 +/- 2.8 mumol/kg FFM x min, P < 0.05). In conclusion losartan improves insulin-mediated glucose uptake through an increase in NOGM and blood flow in hypertensive patients. PMID- 9205939 TI - The effects of atenolol and zofenopril on plasma atrial natriuretic peptide are due to their interactions with target organ damage of essential hypertensive patients. AB - The effects of 10 weeks of treatment with atenolol (n = 9) or the converting enzyme inhibitor zofenopril (n = 25) on plasma atrial natriuretic peptide (ANP) were studied in 34 essential hypertensive patients. After 4 weeks on placebo, pretreatment ANP, 56 +/- 7 pg/ml, was slightly but not significantly higher than that of 29 controls (41 +/- 4) and correlated with age (r = 0.44), ECG score for left ventricular hypertrophy (LVH) (r = 0.51) and serum creatinine (r = 0.67), and negatively with creatinine clearance (r = -0.39). Atenolol reduced blood pressure (BP) by 0 +/- 6/8 +/- 2 mm Hg (ns/P < 0.01), and zofenopril by 14 +/- 4/6 +/- 2 (P < 0.01/P < 0.01), not significantly different between the two agents. Heart rate was decreased by atenolol (-16 +/- 4 bpm, P < 0.01) but not by zofenopril (+1 +/- 2 bpm, ns). Atenolol increased ANP in all patients but one (delta = +42 +/- 9 pg/ml, P < 0.01), while zofenopril did not change it significantly (-6 +/- 6 pg/ml), due to 15 patients exhibiting decreases and 10 increases in plasma ANP. The effect of atenolol on ANP positively correlated with duration of hypertension (r = 0.74), ECG score for LVH (r = 0.73) and serum creatinine (r = 0.68). Individual changes in ANP by zofenopril negatively correlated with pretreatment ANP (r = -0.69), ECG score for LVH (r = -0.44) and serum creatinine (r = -0.41). No correlations were found between BP, heart rate or their changes by treatment and the effect of either agent on plasma ANP. Multiple linear regression showed that the change in ANP was explained by the therapeutic agent used, the pretreatment plasma level of ANP, and the ECG score for LVH (F = 12.5, P < 0.001, r2 = 0.56). We conclude that the effect of antihypertensives on plasma ANP is independent of their action on BP, but dependent on an interaction between the type of drug employed and those clinical characteristics of the patient that reflect pre-existing hypertensive target organ damage. PMID- 9205940 TI - Disparate effects of ACE-inhibitors and calcium antagonists on left ventricular structure and function in essential hypertension. AB - The present study was designed to compare the effects of angiotensin-converting enzyme (ACE) inhibitors and calcium antagonists-the two drug classes thought to be most effective in reducing left ventricular hypertrophy-on arterial pressure, left ventricular structure and function in patients with essential hypertension. After a placebo period of 4 weeks, a population of 96 patients were treated either with one of five different ACE inhibitors or one of six different calcium antagonists. Cardiac structure and function was assessed by 2D-guided M-mode echocardiography. Whereas both drug classes lowered arterial pressure to the same extent, ACE inhibitors had a more pronounced effect on posterior and septal wall thickness and left ventricular mass index than calcium antagonists. Diastolic function, as measured by peak filling rate and duration of rapid filling, improved in both treatment groups to the same extent. However, systolic performance, as assessed by midwall fractional fibre shortening, was significantly improved by ACE inhibitors only. Myocardial contractility (end systolic wall stress/end-systolic volume index) showed no significant change in the ACE inhibitor group but decreased after treatment with calcium antagonists. We conclude that both calcium antagonists and ACE inhibitors lower arterial pressure and increase left ventricular filling to the same extent. However, compared with calcium antagonists, ACE inhibitors had a more pronounced effect on left ventricular mass and improved systolic ventricular performance in patients with essential hypertension. PMID- 9205941 TI - Murine models of human genetic skeletal disorders. AB - Genetic and molecular biological studies have recently resulted in the identification of gene mutations responsible for a large number of human osteochondrodysplasias, skeletal disorders associated with abnormalities of cartilage and/or bone development and growth. Mouse strains carrying mutations in homologous genes are useful, not only because they provide sources of abnormal tissue, but also because specific hypotheses concerning pathogenetic mechanisms can be readily tested. Many of the strains with abnormalities in cartilage and bone have been studied by morphological and biochemical methods for years without successful identification of the genes involved, but recent use of linkage mapping, followed by positional candidate gene cloning, has greatly improved the situation. In a number of instances, the mutations are now known. This is also true for the three "classical" mouse mutants chondrodysplasia (cho), disproportionate micromelia (Dmm) and cartilage matrix deficiency (cmd). In the three strains, mutations in alpha 1(XI) collagen, alpha 1(II) collagen or aggrecan lead to severe defects in the structure and function of cartilage with significant negative effects on longitudinal bone growth. PMID- 9205942 TI - The role of sonic hedgehog in vertebrate development. AB - Members of the hedgehog family are important signalling molecules during embryonic development. One member, Sonic hedgehog, is expressed in embryonic structures such as the zone of polarizing activity in the posterior limb bud, the notochord, and the floor plate of the neural tube, where it plays a role in patterning of the embryo. Sonic hedgehog is synthesized as an inactive precursor which must be proteolytically cleaved and modified by the addition of a cholesterol moiety to become active as a signalling molecule. In this processing, the C-terminal region of Sonic hedgehog serves as both the endoprotease and a cholesterol transferase. The importance of cholesterol for Sonic hedgehog function may explain many of the profound developmental defects caused by perturbations of cholesterol metabolism. The receptor for Sonic hedgehog is Patched, a multi-pass transmembrane protein which forms a complex with Smoothened Mutations in Patched are associated with basal cell naevus syndrome, while mutations in Sonic hedgehog cause holoprosencephaly. Downstream targets of Sonic hedgehog signalling are transcription factors like Gli3, responsible for Greigs polycephalosyndactyly in humans and Hoxd13, responsible for polysyndactyly. PMID- 9205943 TI - Cloning of equine type II procollagen and the modulation of its expression in cultured equine articular chondrocytes. AB - The complete nucleotide sequence of equine type II procollagen has not been previously reported, and equine-specific probes have not been available. We report the complete sequence and discuss the molecular characteristics of equine type II procollagen mRNA which was cloned from a cDNA library prepared from mRNA isolated from equine articular chondrocytes. The coding sequence (4257 bp) was 92.4% homologous to the cDNA of the human sequence, and the propeptide was 97% identical to the human sequence. We demonstrated that when equine chondrocytes are grown in phenotypically-maintained cultures, the expression of type II procollagen is linked to the age of the animal. Additionally, in cultures of young equine chondrocytes, IL1-beta and TNF-alpha both reduced the expression of type II procollagen mRNA in a dose responsive manner. PMID- 9205944 TI - The molecular basis of von Hippel-Lindau disease. PMID- 9205945 TI - The Fas counterattack: a molecular mechanism of tumor immune privilege. PMID- 9205946 TI - Transgenic mice bearing a human mutant thyroid hormone beta 1 receptor manifest thyroid function anomalies, weight reduction, and hyperactivity. AB - BACKGROUND: Resistance to thyroid hormone (RTH) is a syndrome characterized by refractoriness of the pituitary and/or peripheral tissues to the action of thyroid hormone. Mutations in the thyroid hormone receptor beta (TR beta) gene result in TR beta 1 mutants that mediate the clinical phenotype by interfering with transcription of thyroid hormone-regulated genes via a dominant negative effect. In this study, we developed transgenic mice harboring PV, a potent dominant negative human mutant TR beta 1 devoid of thyroid hormone binding and transcriptional activation, as an animal model to understand the molecular basis of this human disease. MATERIALS AND METHODS: Standard molecular biology approaches were used to obtain a cDNA fragment containing mutant PV which was injected into the pronucleus of fertilized egg. Founders were identified by Southern analysis and the expression of PV in tissues was determined by RNA and immunohistochemistry. Thyroid function was determined by radioimmunoassays of the hormones and the behavior of mice was observed using standard methods. RESULTS: The expression of mutant PV was directed by the beta-actin promoter. Mutant PV mRNA was detected in all tissues of transgenic mice, but the levels varied with tissues and with different lines of founders. Thyroid function tests in transgenic mice with high expression of mutant PV showed a significantly (approximately 1.5-fold) higher mean serum total of L-thyroxine levels (p < 0.01) than those of nontransgenic mice. Moreover, thyroid-stimulating hormone levels were not significantly different from those of nontransgenic mice. In addition, these mice displayed decreased weights and a behavioral phenotype characterized by hyperactivity. CONCLUSIONS: These mice have phenotypic features consistent with the commonly observed clinical features of RTH and could be used as a model system to better understand the action of mutant TR beta 1 in a physiological context, which could lead to better treatment for this disease. PMID- 9205947 TI - Human- and mouse-inducible nitric oxide synthase promoters require activation of phosphatidylcholine-specific phospholipase C and NF-kappa B. AB - BACKGROUND: The production of nitric oxide by type II inducible nitric oxide synthase (type II NOS) gene is controlled at least in part by transcriptional activation. Although the murine and human type II NOS genes share significant sequence homology, they differ in the induction stimuli required for activation. MATERIALS AND METHODS: The A549 human and murine RAW 264.7 cell lines were cultured in the presence of inducers of the type II NOS gene and exposed to specific inhibitors of phosphatidyl choline-specific phospholipase C, NF-kappa B, and endocytosis, as well as to reagents that deplete stores of ATP or prevent the acidification of endosomes. The effect of these reagents on the induction of the type II NOS gene transcription, translation, and NO expression was studied using electromobility shift assays, Western blotting, and the detection of NO as nitrates, as appropriate. Additionally, the ability of the native human type II NOS NF-kappa B recognition sequence to bind NF-kappa B was compared with a concensus sequence and with a mutated oligomer. RESULTS: Type II NOS production by both human and mouse cells could be prevented by the addition of the specific inhibitor of phosphatidylcholine-specific phospholipase C, D609, and of agents that interfere with the activation of NF-kappa B. Both mouse and human cells also required acidic endosome formation and the production of 1,2-diacylglycerol for type II NOS expression. Additionally, the native human type II NOS NF-kappa B recognition sequence bound NF-kappa B with significantly less affinity than did the recognition sequence derived from the human immunoglobulin light-chain gene promoter. CONCLUSIONS: These experiments show that whereas mouse cells can be activated by lipopolysaccharide to produce nitric oxide, and human cells require activation by a mixture of cytokines to produce nitric oxide, the intracellular activation pathway following receptor binding of these heterologous stimuli is shared. Additionally, NF-kappa B activation is necessary but not sufficient for inducible nitric oxide synthase production in human cells, in contrast to murine cells in which it serves as a complete inducer. PMID- 9205949 TI - The critical role of p38 MAP kinase in T cell HIV-1 replication. AB - BACKGROUND: Replication of HIV-1 in human T lymphocytes requires the activation of host cellular proteins. This study identifies p38 mitogen-activated protein kinase (MAPK) as one such kinase necessary for HIV-1 replication in T cells. MATERIALS AND METHODS: Primary human T lymphocytes were infected with the LAI strain of HIV-1 and Jurkat cells were infected with the RF strain of HIV-1. HIV replication was measured by reverse transcriptase activity. Cellular expression of endogenous p38 MAPK protein was analyzed using immunoprecipitation. Blockade of p38 MAPK expression was achieved using antisense oligonucleotides to p38 MAPK and the guanylhydrazone compound CNI-1493, an inhibitor of p38 MAPK activation. RESULTS: HIV-1 infection of both primary human T lymphocytes and a T cell line rapidly activated the cellular p38 MAPK pathway, which remained activated for the duration of the culture. Addition of phosphothioated antisense oligonucleotides to p38 MAPK specifically inhibited viral replication. Blockade of p38 MAPK activation by addition of CNI-1493 also inhibited HIV-1 viral replication of primary T lymphocytes in a dose- and time-dependent manner. Stimulation of p38 MAPK activation did not occur with the addition of heat-inactivated virus, suggesting that viral internalization, and not just membrane binding, is necessary for p38 MAPK activation. CONCLUSIONS: These results indicate that activation of the p38 MAPK cascade is critical for HIV-1 replication in primary T lymphocytes, and that blockade of this signal transduction pathway may be a novel therapeutic approach to the treatment of HIV-1 infection. PMID- 9205948 TI - Interleukin-4 receptor expression on AIDS-associated Kaposi's sarcoma cells and their targeting by a chimeric protein comprised of circularly permuted interleukin-4 and Pseudomonas exotoxin. AB - BACKGROUND: AIDS-associated Kaposi's sarcoma (AIDS-KS) represents one of the most common malignancies associated with human immunodeficiency virus infection. To target effective therapeutic agents to AIDS-KS, we have identified a new target in the form of interleukin-4 receptors (IL-4R). MATERIALS AND METHODS: The expression of IL-4R on AIDS-KS cells and their subunit structure was determined by radioligand receptor binding, cross-linking and Northern and RT-PCR analyses. The in vitro effect of IL-4 and recombinant fusion protein made up of circularly permuted IL-4 and a mutated form of Pseudomonas exotoxin, IL-4(38-37)-PE38KDEL, was examined by clonogenic and protein synthesis inhibition assays. RESULTS: Five AIDS-KS cell lines expressed high-affinity IL-4R with a Kd of 23.5-219 pM. IL-4 appeared to cross-link to one major protein corresponding to 140 kDa and a broad band corresponding to 60-70 kDa. Both cross-linked proteins were immunoprecipitated with an antibody to human IL-4R beta chain. AIDS-KS cells exhibited IL-4R beta-specific mRNA. IL-4 caused a modest inhibition (31-34%) of colony formation in two AIDS-KS cell lines tested. IL-4(38-37)-PE38KDEL was found to be highly effective in inhibiting the protein synthesis in all five AIDS-KS examined. The IC50 ranged from 32 to 1225 pM. The cytotoxic action of IL-4 toxin was blocked by an excess of IL-4, exhibiting the specificity of IL-4(38-37) PE38KDEL. The cytotoxicity of IL-4 toxin observed by a clonogenic assay corroborated well with the IC50 obtained by protein synthesis inhibition assay. Normal human endothelial cells expressed a negligible number of IL-4R (< 50 sites/cell) and were less sensitive or not sensitive to IL-4(38-37)-PE38KDEL. CONCLUSION: The presence of a new plasma membrane protein in the form of IL-4R on AIDS-KS cells may be targeted by IL-4(38-37)-PE38KDEL for its potential implication in the treatment of AIDS-KS. PMID- 9205950 TI - Modulation of human cardiac function through 4 beta-adrenoceptor populations. AB - In human heart there is now evidence for the involvement of four beta adrenoceptor populations, three identical to the recombinant beta 1-, beta 2- and beta 3-adrenoceptors, and a fourth as yet uncloned putative beta-adrenoceptor population, which we designate provisionally as the cardiac putative beta 4 adrenoceptor. This review described novel features of beta-adrenoceptors as modulators of cardiac systolic and diastolic function. We also discuss evidence for modulation by unoccupied beta 1- and beta 2-adrenoceptors. Human cardiac and recombinant beta 1- and beta 2-adrenoceptors are both mainly coupled to adenylyl cyclase through Gs protein, the latter more tightly than the former. Activation of both human beta 1- and beta 2-adrenoceptors not only increases cardiac force during systole but also hastens relaxation through cyclic AMP-dependent phosphorylation of phospholamban and troponin 1, thereby facilitating diastolic function. Furthermore, both beta 1 and beta 2-adrenoceptors can mediate experimental arrhythmias in human cardiac preparations elicited by noradrenaline and adrenaline. Human ventricular beta 3-adrenoceptors appear to be coupled to a pertussis toxin-sensitive protein (Gi?). beta 3-Adrenoceptor-selective agonists shorten the action potential and cause cardiodepression, suggesting direct coupling of a Gi protein to a K+ channel. In a variety of species, including man, cardiac putative beta 4-adrenoceptors mediate cardiostimulant effects of non conventional partial agonists, i.e. high affinity beta 1- and beta 2-adrenoceptor blockers that cause agonist effects at concentrations considerably higher than those that block these receptors. Putative beta 4-adrenoceptors appear to be coupled positively to a cyclic AMP-dependent cascade and can undergo some desensitisation. PMID- 9205951 TI - Agonist activity of antimigraine drugs at recombinant human 5-HT1A receptors: potential implications for prophylactic and acute therapy. AB - The actions of several serotonergic ligands in use or under development for the treatment of migraine headaches were examined at recombinant human 5-HT1A receptors stably expressed in Chinese Hamster Ovary cells. Affinities (K(i)s) at this site were determined in competition binding experiments with [3H]-8-OH-DPAT ([3H](+/-)8-hydroxy-N,N-dipropylaminotetralin), whilst agonist efficacy was measured by stimulation of [35S]-GTP gamma S (guanylyl-5'-[gamma[35S]thio] triphosphate) binding. Of the prophylactic antimigraine drugs tested, methysergide and lisuride behaved as efficacious agonists (Emax > or = 90% relative to 5-HT) whereas pitozifen and (-)propranolol acted as a partial agonist (60%) and an antagonist, respectively. This suggests that there is no correlation between agonism at 5-HT1A receptors and prophylactic antimigraine action. In contrast, serotonin, dihydroergotamine, sumatriptan, naratriptan and alniditan, which are effective in acute interruption of migraine attacks, each displayed high efficacy (Emax = 100, 100, 92.6, 79.3, 79.1% respectively) and marked affinity (Ki = 18.7, 0.6, 127, 26.4 and 3.0 nM respectively) at 5-HT1A receptors. EC50 values for agonist stimulation of [35S]-GTP gamma S binding correlated with respective Ki values at 5-HT1A receptors (r = 0.93) and the stimulation of [35S] GTP gamma S binding by these compounds was antagonised by the selective 5-HT1A antagonist WAY 100,635 (N-{2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl}-N-(2 pyridinyl) cyclo-hexanecarboxamide; 100 nM). These data suggest that agonism at 5 HT1A receptors may be involved in some actions of drugs used in acute antimigraine therapy. In comparison with the above compounds, novel ligands targeted at 5-HT1B/1D receptors, such as GR125,743 (N-[4-methoxy-3-(4-methyl piperazin-1-yl)phenyl] -3-methyl-4-(4-pyridyl)benzamide) and GR 127,935 (N-[4 methoxy-3-(4-methylpiperazin-1-yl)-phenyl]-2'-methyl-4'-(5-m ethyl-1, 2,4 oxadiazol-3-yl)-biphenyl-4-carboxamide), only weakly activated [35S]-GTP gamma S binding (32.4 and 32.1% efficacy) and displayed moderate affinity at 5-HT1A receptors (Kis 53.1 and 49.8 nM) suggesting that they constitute useful tools to differentiate 5-HT1A and 5-HT1B/1D receptor-mediated actions. In conclusion, the present data indicates that several antimigraine agents exhibit marked 5-HT1A receptor activity and that although this is unlikely to be important for prophylactic action it may be relevant to the ancilliary properties of drugs used for acute migraine treatment. PMID- 9205952 TI - Celiprolol: agonist and antagonist effects at cardiac beta 1- and vascular beta 2 adrenoceptors determined under in vivo conditions in the rat. AB - Celiprolol is a beta-adrenoceptor antagonist which has desirable ancillary properties since it is relatively cardioselective and can exert direct vasodilator and bronchodilator effects. Here agonist and antagonist effects of celiprolol at cardiac beta 1- and vascular beta 2-adrenoceptors were determined under in vivo conditions in the rat. All experiments were carried out in catecholamine-depleted, pentobarbital anesthetized and vagotomized rats, placed under artificial respiration. I.v. administrations were made via the femoral vein. Blood pressure was measured from the cannulated right carotid artery and heart rate was recorded with a cardiotachometer. Celiprolol (10 micrograms/kg to 1 mg/kg i.v.) produced dose-related increases in heart rate and decreases in mean carotid artery blood pressure which were of longer duration than those mediated by standard agonists of beta 1-(isoprenaline) or beta 2-(salbutamol) adrenoceptors respectively. Although the maximal increase in heart rate by celiprolol (110 +/- 4 beats/min, n = 7) was approximately half that of isoprenaline (198 +/- 1 beats/min, n = 5), isoprenaline acted at doses 200-fold lower than celiprolol. Betaxolol (0.03-0.3 mg/kg i.v.), a beta 1-adrenoceptor antagonist, inhibited strongly and with similar potency the tachycardiac effects of celiprolol (DR10 = 45 micrograms/kg i.v.) as well as isoprenaline (DR10 = 45 micrograms/kg i.v.). On the other hand, the hypotensive effects of celiprolol and salbutamol were antagonized markedly and with similar potency by ICI 118,551, a relatively selective beta 2-adrenoceptor antagonist (DR10 = 15 and 25 micrograms/kg i.v. respectively). In rats pretreated with celiprolol (0.03 to 0.3 mg/kg i.v.), the heart rate dose-response curves to isoprenaline were shifted to the right of those determined in matched groups of vehicle-pretreated animals. In this respect, celiprolol was half as potent as betaxolol in blocking cardiac beta 1-adrenoceptors. Furthermore, celiprolol also antagonized the hypotensive effects of salbutamol, but, in this respect, celiprolol was 90-fold less potent than ICI 118,551. In conclusion, these results clearly indicate that celiprolol has the ability of stimulating and blocking not only cardiac beta 1- but also vascular beta 2-adrenoceptors. The effects on cardiac beta 1-adrenoceptors as well as the agonism of beta 2-adrenoceptors are produced by similar doses of celiprolol. These doses are notably lower than those necessary to block beta 2-adrenoceptors. Thus, this pharmacological profile, which has also been demonstrated in humans, indicates that celiprolol is a modulator of cardiac beta 1-adrenoceptors with vascular beta 2-adrenoceptor agonist properties. PMID- 9205953 TI - Lanthanides inhibit the human noradrenaline, 5-hydroxytryptamine and dopamine transporters. AB - Transporters for the monoamine neurotransmitters, including noradrenaline, 5 hydroxytryptamine [5-HT] and dopamine, have twelve transmembrane spanning regions and cotransport Na+ and Cl- ions. Another family of Na(+)-dependent transporters is that containing the Na+/glucose and Na+/proline cotransporters that are found in the epithelial cells of renal and intestinal brush border membranes. It has been shown that various trivalent lanthanides can substitute for Na+ for transport of glucose and proline. The aim of this study was to determine the effects of lanthanides on the activities of the human noradrenaline, 5-HT and dopamine transporters. Cultured cells were incubated for 2 min with 10 nM 3H noradrenaline (SK-N-SH-SY5Y human neuroblastoma cells), 3H-5-HT (JAR human placental choriocarcinoma cells) or 3H-dopamine (COS-7 cells transfected with the cDNA of the human dopamine transporter). Specific amine uptake was determined as the difference between accumulation of the amine in the cells in the absence and presence of a corresponding uptake inhibitor. Under both isotonic (150 mM NaCl or LiCl or 90 mM lanthanide salt) and hypertonic (150 mM NaCl +100 mM LiCl, 250 mM LiCl or 150 mM lanthanide salt) conditions, replacement of Na+ by Li+, La3+, Eu3+ or Sm3+ abolished the specific uptake of noradrenaline in SK-N-SH-SY5Y cells and replacement of Na+ by Li+ or Eu3+ decreased the specific uptake of 5-HT in JAR cells by 94-100% and that of dopamine in transfected COS-7 cells by 95-99%. The direct effects of Eu3+ (with Na+ present) on the human noradrenaline transporter in SK-N-SH-SY5Y cells were also examined. Eu3+ inhibited noradrenaline uptake into the cells (IC50 2.6 mM) and nisoxetine binding to crude membranes of SK-N-SH SY5Y cells (IC50 4.7 mM) with similar potencies. Further experiments showed that 4.5 mM EuCl3 in the presence of 150 mM Na+ caused a 3.5-fold increase in the K(m) of noradrenaline and no change in the maximal rate of noradrenaline uptake. EuCl3 (4.5 mM) also caused a pronounced inhibition of the Na(+)-dependent stimulation of noradrenaline uptake by SK-N-SH-SY5Y cells. It can be concluded from these data that, in contrast with the Na+/glucose and Na+/proline cotransporters, the lanthanides cannot substitute for Na+ in the transport of substrates by the monoamine neurotransmitter transporters and that the lanthanides inhibit the latter transporters by interacting with sites of the transporters involved in amine and Na+ binding. PMID- 9205955 TI - On the involvement of a tonic dopamine D2-autoinhibition in the regulation of pulse-to-pulse-evoked dopamine release in the rat striatum in vivo. AB - The dopamine overflow evoked by trains of electrical stimulation pulses applied to the ascending dopaminergic pathway was measured with continuous amperometry in the striatum of anesthetised rats. As previously observed in in vitro studies, a pulse by pulse analysis showed a fall in dopamine overflow evoked by pulses 2 to 6, compared to the response evoked by pulse 1. However, in contrast with in vitro findings, the present in vivo data showed that the dopamine receptor antagonist haloperidol i) completely reverses the fall in dopamine overflow between pulse 1 and subsequent pulses, ii) enhances the dopamine overflow elicited by pulse 1. These results suggest that in vivo, both basal and pulse-evoked dopamine overflow results in stimulation of dopamine D2-type autoreceptors and therefore in regulation of dopamine release. PMID- 9205954 TI - P2-receptor-mediated inhibition of noradrenaline release in the rat hippocampus. AB - Experiments on hippocampal slices were carried out in order to find out whether the release of noradrenaline in the hippocampus can be modulated through P2 receptors. The slices were preincubated with [3H]-noradrenaline, superfused with medium containing desipramine (1 microM), and stimulated electrically, in most experiments by 4 pulses/100 Hz. The adenosine A1-receptor agonist N6-cyclopentyl adenosine (CPA) and the nucleotides ATP, adenosine-5'-O-(3-thiotriphosphate) (ATP gamma S) and adenosine-5'-O-(2-thiodiphosphate) (ADP beta S) decreased the evoked overflow of tritium by up to 55%. The adenosine A2a-agonist 2-p-(2-carboxyethyl) phenethylamino-5'-N-ethylcarboxamido-adenosin e (CGS 21680; 0.003-0.3 microM) caused no change. The concentration-response curve of CPA was shifted to the right by the A1-antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX; 3 nM) but not by the P2-receptor antagonists cibacron blue 3GA (30 microM) and reactive blue 2 (30 microM); the apparent pKB value of DPCPX against CPA was 9.0. In contrast, the concentration-response curve of ATP was shifted to the right by DPCPX (3 nM), apparent pKB 8.7, as well as by cibacron blue 3GA (30 microM), apparent pKB 5.2, and reactive blue 2 (30 microM), apparent pKB 5.6; the antagonist effects of DPCPX and cibacron blue 3GA were additive in a manner compatible with the blockade of two separate receptors for ATP. The same pattern was obtained with ATP gamma S: its concentration-response curve was shifted to the right by DPCPX as well as by cibacron blue 3GA and reactive blue 2. Suramin (300 microM) antagonized neither the effect of ATP nor that of ATP gamma S. The 5'-nucleotidase inhibitor alpha, beta-methylene-ADP (100 microM) did not change the effect of ATP. Only cibacron blue 3GA (30 microM) but not reactive blue 2 (30 microM), given alone, consistently caused a small increase of the evoked overflow of tritium. Hippocampal slices degraded exogenous ATP, and this degradation was reduced by cibacron blue 3GA (30 microM), reactive blue 2 (30 microM) and suramin (300 microM). The results indicate that the noradrenergic terminal axons of the rat hippocampus possess P2-receptors in addition to the known A1-adenosine receptors. The presynaptic P2-receptors mediate an inhibition of noradrenaline release, are activated by nucleotides but not nucleosides, and are blocked by cibacron blue 3GA and reactive blue 2. ATP and ATP gamma S act at both the A1- and the P2-receptors. An autoreceptor function of cerebral presynaptic P2 receptors remains doubtful. PMID- 9205956 TI - Influence of blockade of alpha 1-adrenoceptors, beta 1-adrenoceptors and vasopressin V1A receptors on the cardiovascular effects of gamma 2-melanocyte stimulating hormone (gamma 2-MSH). AB - gamma 2-Melanocyte-stimulating hormone (gamma 2-MSH) and related melanotropins have been shown to have various cardiovascular effects, including acute, short lasting increases in blood pressure (MAP) and heart rate (HR). gamma 2-MSH, administered intravenously, dose-dependently increased MAP and HR with an ED50 of approximately 30 nmol/kg and a maximal effect on MAP of approximately 55 mm Hg and on HR of around 70 beats per minute. Intravenous (i.v.) pretreatment with the alpha 1-adrenoceptor antagonist, prazosin, caused the dose-response curve for the effect of gamma 2-MSH on MAP to shift to the right with a decrease in slope, whereas it had no effect on the dose-response curve for the effect on HR. I.v. pretreatment with the beta 1-adrenoceptor antagonist, metoprolol, had no effect on the dose-response curve for the effect of gamma 2-MSH on MAP, but it caused the dose-response curve for the effect of the peptide on HR to shift to the right with a decrease in slope. Neither i.v. nor intracerebroventricular (i.c.v.) administration of the vasopressin V1A receptor antagonist, SR 49059 ((2S) 1-[(2R 3S)-5-chloro-3-(2-chlorophenyl)-1-(3,4-dimethoxy-benzene-sulfonyl)- 3-hydroxy-2,3 dihydro-1H-indole-2-carbonyl]-pyrrolidine-2-carboxamide), had significant effects on the dose-response curves for the effects of the peptide on either MAP or HR. The doses of prazosin, metoprolol and SR 49059 were found to be effective in counteracting the effects of agonists for these receptors (phenylephrine, isoprenaline and [Arg8]vasopressin, respectively). Taken together, these results support the postulate that the effects of gamma 2-MSH are, at least partially, due to an increase in sympathetic outflow to the periphery (Gruber and Callahan (1989), Am J Physiol 257: R681-R694), and that this increase leads to increased activation of vascular alpha 1-adrenoceptors and cardiac beta 1-adrenoceptors. If, as was suggested by these authors, gamma 2-MSH acts via activation of a central vasopressin system, it is via a vasopressin receptor subtype other than the vasopressin V1A receptor, since i.c.v. administration of a selective vasopressin V1A receptor antagonist failed to interfere with the pressor and cardioaccelerator effects of gamma 2-MSH. PMID- 9205957 TI - Na+ channel modulation and force-frequency relationship in human myocardium. AB - The enhancement of force of contraction (FOC) following increasing frequencies of stimulation is an important mechanism of positive inotropy in human myocardium. The present study aimed to investigate the influence of alterations in Na+ influx on FFR in human myocardium. Isometric FOC of electrically stimulated right auricular trabeculae (AUT, n = 12) from human non-failing hearts (n = 8) was measured at different stimulation rates (0.5-3 Hz) under control conditions, after increasing Na+ influx by the addition of (+/-)BDF 9148 (BDF, 3 mumol l-1) and after decreasing Na+ influx by the addition of lidocaine (LIDO, 10 mumol l 1). Additionally, the rate dependent changes in diastolic tension (DT) were measured in all experiments. Under control conditions FOC increased with increasing frequencies of stimulation. The rate at which maximal FOC was observed (SFmax) was 2.0 +/- 0.2 Hz and maximal increase in FOC (PIEmax) by increasing frequency of stimulation was +1.5 +/- 0.5 mN. After increase of Na+ influx by BDF (3 mumol l-1) SFmax was decreased to 0.8 +/- 0.1 Hz (p < 0.05 versus control) and PIEmax was +0.1 +/- 0.3 mN (p < 0.05). When Na+ influx was diminished by LIDO (10 mumol l-1) SFmax and PIEmax were increased compared to control (2.4 +/- 0.1 Hz and +4.1 +/- 0.9 mN, p < 0.05 versus control). The diastolic tension (DT) of AUT at 3 Hz was not changed at higher rates in the control group and after application of LIDO (10 mumol l-1), whereas after enhancement of Na+ influx by BDF there was an increase in DT of +0.7 +/- 0.2 at 3 Hz (p < 0.05 versus control and LIDO). An enhanced Na+ influx leads to a decrease in the optimal frequency and to a smaller force potentiation by higher stimulation rates which could be at least partly due to incomplete relaxation at higher frequencies, whereas a reduced Na+ influx is followed by opposite alterations. It is concluded that besides Ca2+ handling also Na+ influx and Na+ homeostasis might determine the frequency-induced force potentiation in human myocardium. Thus, the negative FFR in diseased human myocardium might result from changes in cellular Ca2+ or Na+ regulatory sites. PMID- 9205958 TI - Stereoselectivity of actions of the calcium sensitizer [+]-EMD 60263 and its enantiomer [-]-EMD 60264. AB - The thiadiazinone derivative [+]-EMD 60263 ((+)-5-(l-(alpha-ethylimino-3,4 dimethoxybenzyl)-1,2,3,4-tetrah ydroquinoline -6-yl)-6-methyl-3,6-dihydro-2H 1,3,4 -thiadiazine-2-on) is a Ca(2+)-sensitizing agent with only minor phosphodiesterase inhibitory activity. Our aim was to characterize the inotropic and electrophysiological effects of [+]-EMD 60263 and its enantiomer [-]-EMD 60264 in several cardiac muscle preparations. The Ca(2+)-sensitizing activity resided in the [+]-enantiomer only. [+]-EMD 60263 (3 microM) shifted the EC50 of Ca2+ for contractile activation of skinned fibers of pig heart from 2.41 microM to 0.73 microM, whereas [-]-EMD 60264 (30 microM) was ineffective. In Langendorff perfused guinea pig hearts, [+]-EMD 60263 and [-]-EMD 60264 induced concentration dependent positive and negative inotropic effects, respectively; both enantiomers reduced spontaneous heart rate but did not influence perfusion pressure. The maximum increase in force of human atrial trabeculae was 35% of pre-drug control with [+]-EMD 60263 in comparison to 113% with forskolin. In guinea-pig papillary muscles, [+]-EMD 60263 and [-]-EMD 60264 had opposite inotropic responses, however, both agents similarly prolonged action potential duration. Both enantiomers concentration-dependently blocked the rapidly activating component IKr of the delayed rectifier in guinea-pig myocytes. The block saturated at potentials positive to +30 mV, closely resembling the effects of the antiarrhythmic agent E-4031 which had been originally used to define IKr. PMID- 9205959 TI - Blockade of the human cardiac K+ channel Kv1.5 by the antibiotic erythromycin. AB - Erythromycin administration has been associated with a prolongation of cardiac repolarization in certain clinical settings. This could be due to blockade of voltage-dependent K+ channels in the human heart. For this reason we examined the effects of erythromycin on a rapidly activating delayed rectifier K+ channel (Kv1.5) cloned from human heart and stably expressed in human embryonic kidney cells. When examined using the whole-cell patch clamp technique, erythromycin (100 microM) blocked Kv1.5 current in a time-dependent manner but required prolonged exposure to do so. However, when we examined Kv1.5 current using inside out macro-patches, erythromycin applied to the cytoplasmic surface rapidly (within 1-2 min) inhibited Kv1.5 current with an IC50 value of 2.6 x 10(-5)M (1.7 - 3.9 x 10(-5)M, 95% C.L.). The main effect of erythromycin was to accelerate the rate of Kv1.5 current decay thereby reducing the current at the end of a prolonged voltage-clamp pulse. Erythromycin also blocked Kv1.5 current in both a voltage- and frequency-dependent manner but had little effect on the activation kinetics, deactivation kinetics, or the steady-state inactivation properties of Kv1.5. These data suggest that erythromycin acts as a blocker of an activated state of the Kv1.5 channel and that it may access its binding site from the intracellular face of the channel. This study is the first to examine the effects of erythromycin on a cloned human cardiac K+ channel. It is concluded that erythromycin blocks Kv1.5 at clinically relevant concentrations. Blockade of voltage-dependent K+ channels in the heart could contribute to the alterations in cardiac repolarization that have been observed with erythromycin. PMID- 9205960 TI - Palytoxin-induced increase in endothelial Ca2+ concentration in the rabbit aortic valve. AB - Palytoxin (PTX) is one of the most potent toxins isolated from marine coelenterates of the genus Palythoa. It induces depolarization in various types of cells by increasing the permeability for monovalent cations. It has been reported that PTX induces endothelium-dependent relaxation of vascular smooth muscle. In this study, we examined the effect of PTX on the cytosolic Ca2+ concentration ([Ca2+]i) in the endothelium of rabbit aortic valves loaded with fluorescent Ca2+ indicators, fura-PE3 or fluo-3. PTX (10 pM-300 nM) irreversibly increased endothelial [Ca2+]i in a concentration-dependent manner. ATP and thapsigargin also increased [Ca2+]i. Imaging of [Ca2+]i with a confocal microscope revealed that PTX increased [Ca2+]i in all endothelial cells studied (n = 13). An inorganic Ca2+ entry blocker, La3+ (30 microM), had no effect on the increase in [Ca2+]i induced by PTX whereas it inhibited the sustained phase of the increase in [Ca2+]i induced by ATP or thapsigargin. The PTX-induced increase in [Ca2+]i was partially inhibited by ouabain and was abolished by removal of external Ca2+ although decrease of Na+ concentration in the incubation medium was ineffective. Activation of protein kinase C by 1 microM 12-deoxyphorbol 13 isobutyrate or inhibition of phosphatase by 10 nM calyculin-A had no effect on the increase in [Ca2+]i induced by PTX, whereas both agents inhibited the sustained phase of the increase in [Ca2+]i induced by ATP or thapsigargin. Mn2+ influx, measured by the quenching of fura-PE3 fluorescence, was accelerated by ATP or thapsigargin, but not by PTX. These results suggest that PTX increases [Ca2+]i in the endothelium of the rabbit aortic valve by increasing Ca2+ influx through a pathway which is different from that activated by ATP or thapsigargin. PMID- 9205961 TI - Functional classification of P1-purinoceptors in guinea-pig left and right atria: anomalous characteristics of antagonism by cyclopentyltheophylline. AB - The P1 purinoceptor subtype mediating the negative inotropic responses of guinea pig left atria and the negative chronotropic responses of beating right atria were characterized. Guinea-pig isolated paced left atria (2Hz, 5ms, threshold voltage+50%) and spontaneously beating right atria were set up in Krebs bicarbonate solution and isometric tension and rate of contraction, respectively, were recorded. Concentration-response curves for the reduction of tension and rate, respectively, by adenosine receptor agonists, N6-cyclopentyladenosine (CPA), the R- and S- stereoisomers of N6-(2-phenylisopropyl) adenosine (R-PIA and S-PIA), 5'-(N-carboxamido) adenosine (NECA) and 2-p-((carboxyethyl) phenethylamino)-5'-(N-carboxamido) adenosine (CGS21680) were obtained. The orders of potency on the left atria (CPA = NECA > R-PIA > S-PIA > CGS21680) and right atria (CPA = R-PIA > S-PIA > CGS21680) were consistent with the responses being mediated via A1 receptors. Antagonism of the responses to CPA or R-PIA by 8-cyclo 1,3-dimethylxanthine (CPT) was examined by a full Schild analysis. Concentration response curves for CPA or R-PIA were obtained in the absence or presence of five or six concentrations (10(-7)-10(-5) or 3 x 10(-5)M) of CPT. The shift in the concentration-response by CPT was expressed as the concentration-ratio (CR) and plotted as -log(CR-1) against log molar concentration of CPT (Schild plot). pA2 values were calculated from the intercept on the concentration axis and by application of the equation; pA2 = log(antagonist concentration) -log (CR-1). The Schild plots had unity slopes indicating competitive antagonism and the pA2 values derived therefrom indicated that the responses were mediated via A1 receptor. Closer inspection of the Schild plots, however, showed that at the higher concentrations of CPT there was a limit to the displacement of the concentration-response curves of the left and right atria to CPA and of the left atria to R-PIA. There were also significant differences in the apparent pA2 values calculated from the equation, when different concentrations of antagonist were examined. These results indicated that at higher concentrations of agonist there may be a component of the response that is resistant to antagonism by CPT. Whether this is related to the proposal that cardiac responses are mediated via A3 receptors is discussed. PMID- 9205962 TI - Behavioural pharmacology of the non-competitive NMDA antagonists dextrorphan and ADCI: relations between locomotor stimulation, anticataleptic potential and forebrain dopamine metabolism. AB - The effects of systemic administration of the non-competitive N-methyl-D aspartate (NMDA) antagonists dextrorphan (10-40 mg/kg, i.p.) and [+/-]-5 aminocarbony-10,11-dihydro-5H-dibenzo[a,d]cycloheptan++ +-5,10-imine (ADCI) (25 70 mg/kg, i.p.) on basal ganglia-mediated behaviour and on forebrain dopamine metabolism were investigated in rats. Dextrorphan increased locomotor activity but did not induce stereotyped sniffing. ADCI failed to produce any significant motor stimulant and motor depressant actions. Both dextrorphan and ADCI dose dependently antagonized catalepsy induced by the D-1 dopamine receptor antagonist SCH 23390 or the D-2 dopamine receptor antagonist haloperidol. Only the highest doses of dextrorphan and ADCI increased dopamine metabolism in the prefrontal cortex and/or in the nucleus accumbens, but not in the dorsal striatum. Our results show that dextrorphan and ADCI produce some of the behavioural effects (antagonism of experimentally induced catalepsy) and neurochemical actions (regionally selective stimulation of dopamine metabolism) that have previously been observed in the prototypical non-competitive NMDA antagonist, dizocilpine. The failure of ADCI to induce hyperlocomotion and stereotypy suggests that anticataleptic doses of ADCI may be devoid of the psychotomimetic actions commonly associated with non-competitive blockade of NMDA receptor function. PMID- 9205963 TI - Daily rhythms of heart rate, temperature and locomotor activity are modified by anaesthetics in rats: a telemetric study. AB - The aim of this study was to evaluate the effect of anaesthesia (ether or ketamine) on daily rhythms of temperature (T), heart rate (H) and locomotor activity (A) in unrestrained rats by using implanted radio-telemetry transmitters. T, H and A were measured every 10 min, in Wistar male rats, and analysed using Cosinor. The mean +/- SEM days needed, after surgical implantation, to detect a daily rhythm in H, T and A were also assessed. Six rats were anaesthetized for about 50 min either by ketamine or ether in a 3 by 3 cross over design. Mesors, amplitudes and acrophases of T, H and A were calculated three days before (D-3; D-2; D-1), the day of anaesthesia (D0) as well as the three following days (D1; D2; D3). ANOVA was performed in order to detect, firstly a possible effect due to the order of application of anaesthesia, secondly a significant difference between ether or ketamine-induced anaesthesia and finally a modification of the mesors, amplitudes and acrophases of T, H and A, induced by each anaesthesia, for D0, D1, D2 and D3 when compared to D-1. Our results indicate: (1) Alterations of the acrophases, mesors and amplitudes, except for the amplitude of A, of the daily rhythms of T, H and A on D0 of ketamine anaesthesia while regarding ether anaesthesia only amplitude of T and H and acrophase of A were modified on D0. Some of these modifications were still observed on the days following anaesthesia. A significant difference between ether and ketamine-induced anaesthesia was also observed. (2) A non-detection of T, H and A daily rhythms after surgical implantation, which was not observed after injection of either ether or ketamine alone. Almost 10 days were needed to detect a significant daily rhythm for T, H and A. The authors suggest that, the general anaesthetic agent was responsible for a perturbation of the mesors, amplitudes and acrophases of the daily rhythms of H, T and A while the non detection of these rhythms after implantation was more due to the surgical aggression. PMID- 9205964 TI - Effects of L-701,324, a high-affinity antagonist at the N-methyl-D-aspartate (NMDA) receptor glycine site, on the rat electroencephalogram. AB - L-701,324 (7-chloro-4-hydroxy-3-(3-phenoxy) phenyl-2-(1H)-quinolone) is a novel, orally active antagonist at the N-methyl-D-aspartate (NMDA) receptor glycine site. As NMDA receptor antagonism is generally associated with anaesthetic effects, we have examined the electroencephalographic alterations produced by doses of L-701,324 that effectively reduce NMDA-evoked responses in vivo. Microdialysis probes incorporating an electrode were implanted in the striatum of rats and perfused with artificial cerebrospinal fluid (ACSF). Under light halothane anaesthesia, 12 consecutive depolarizations were elicited by switching to ACSF containing 200 microM NMDA for 2 or 3 min, every 20 min. NMDA-evoked depolarizations and EEG were recorded with the microdialysis electrode. L-701,324 (5 or 10 mg kg-1 i.v.) or vehicle were administered 5 min after the 3rd NMDA stimulus. L-701,324 dose-dependently inhibited NMDA-evoked depolarizations, with 10 mg kg-1 reducing these responses by 50% for at least 3 h. The average amplitude of the EEG in the window 0.25-6 Hz (low frequencies) and 6-21 Hz (high frequencies) did not change in the control group. At the higher dose of 10 mg kg 1 L-701,324 transiently increased the amplitude of low frequencies by around 20%. In contrast, both 5 and 10 mg kg-1 significantly reduced the high frequencies to around 70% of control, and this action was sustained with the higher dose. Analysis of the relative EEG power spectra confirmed a small, but persistent shift from high to low EEG frequencies. Our results suggest that L-701,324 slightly strengthened halothane anaesthesia at doses inhibiting effectively NMDA receptor function. Accordingly, the resulting anticonvulsant and neuroprotective actions of L-701,324 may not be associated with marked anaesthesia-like side effects. PMID- 9205965 TI - Stimulation of bladder activity by volume, L-dopa and capsaicin in normal conscious rats--effects of spinal alpha 1-adrenoceptor blockade. AB - To study possible differences in alpha 1-adrenoceptor involvement in the spinal mechanisms mediating bladder activity induced by volume (bladder filling), central (L-dopa), and peripheral (capsaicin) stimulation, we investigated if these types of bladder activity were modified by intrathecal (i.t.) or intra arterial (i.a.) administration of the alpha 1-adrenoceptor antagonist, indoramin. Indoramin is selective for the alpha 1A-adrenoceptor subtype, whereas most clinically used alpha 1-adrenoceptor antagonists, including doxazosin, have no subtype selectivity. The drug effects were studied by continuous cystometry in normal, conscious rats and rats with bladder activity evoked by intraperitoneal L dopa (50 mg/kg after carbidopa pretreatment), or by intravesical capsaicin (30 microM). I.t. indoramin (50 nmol) significantly decreased micturition pressure, and increased bladder capacity and micturition volume. Dribbling incontinence due to urinary retention was observed in one of ten rats. L-dopa-stimulated bladder overactivity was significantly attenuated by i.t. or i.a. indoramin (50 nmol). Similar effects of i.t. and i.a. doxazosin (50 nmol) have been reported previously. Intravesical capsaicin (30 microM) caused bladder activity, which was attenuated by i.t. indoramin (50 nmol), but not by i.t. doxazosin (50 nmol). I.a. indoramin did not reduce capsaicin-induced bladder activity; doxazosin was moderately effective. The results suggest that the bulbospinal micturition reflex evoked by bladder filling and L-dopa involves a descending pathway where transmission is partly mediated by spinal alpha 1-adrenoceptors. Bladder overactivity evoked by intravesical capsaicin, which elicits a vesicospinal vesical reflex, was not affected by i.t. doxazosin in a dose that attenuates activity mediated through the bulbo-spinal pathway. This suggests less involvement of spinal alpha 1-adrenoceptors in the vesico-spinal-vesical than in the bulbo-spinal voiding reflex. PMID- 9205966 TI - Side effects of adjuvant chemotherapy: perceptions of node-negative breast cancer patients. AB - Twenty-one node-negative breast cancer patients were interviewed shortly after completing adjuvant chemotherapy and asked about side effects they had experienced, expectation of side effects, and strategies for coping with the side effects. Eighteen of the women were interviewed 6 months later to determine their feelings about the chemotherapy experience and ending treatment and what side effects persisted or developed after chemotherapy. Hair loss, fatigue, treatment related problems, nausea and infections/low blood counts were the most frequently described problems during the first interviews. Patients used coping strategies suggested by physicians and nurses. Six months later, hair problems, fatigue, weight gain, menopausal problems, emotional problems and nail problems were most often reported. Most patients (16/18) did not expect to be experiencing chemotherapy-related problems 6 months after ending treatment. Fatigue interfered with daily lives and weight gain caused concern. A total of 35% of participants experienced fear or anxiety at the end of chemotherapy, but most (62%) recalled at least some positive feelings 6 months later. Given the same circumstances, all but two would make the same decision to undergo adjuvant chemotherapy. Support groups would be especially useful for patients completing chemotherapy who would lose continued frequent support from clinic personnel. PMID- 9205967 TI - Fear of cancer recurrence--a literature review and proposed cognitive formulation to explain exacerbation of recurrence fears. AB - Modern treatments for cancer are resulting in cancer patients living longer with the risk of the disease returning at a later stage. Many patients who experience a recurrence blame themselves (Mahon et al., 1990), while those in remission live with the constant fear that the cancer may return. Although fear of recurrence is recognised in the literature, few researchers have focused on this aspect as a precursor to psychological distress. This paper reviews the literature about fear of recurrence and its measurement. Leventhal's Self Regulation Model of Illness is presented to help understand patients' reactions to cancer and fears for the future. The authors propose a formulation for fear of recurrence and examples of interventions that are indicated. PMID- 9205968 TI - A preliminary survey of oncologists' perceptions of quality of life information. AB - Quality of life (QOL) of cancer patients has become the focus of increasing research in oncology, and a frequently measured endpoint in clinical trials. Very little attention has been paid to the perspective of physicians on quality of life information, and its role in clinical decision-making. This report describes the findings of research focused on exploring the perspectives of physicians about quality of life information that is available for cancer patients. On the basis of qualitative data gathered through in-depth interviews with 60 oncologists in the first phase of this project, we have developed a self administered questionnaire (MD-QOL survey) designed to assess oncologists' views on QOL. This survey was administered to an international group of gynecologic oncologists. The objectives of this study were to assess the face validity of the initial items in the MD-QOL, to expand the pool of items, and to assess the feasibility of utilizing a self-administered questionnaire to assess physicians' perspectives on QOL information. Twenty-eight oncologists responded to the questionnaire. The majority of respondents felt that QOL can be measured and that it should be measured from the patient's own perspective. Half of the physicians felt that currently available QOL information is useful in clinical practice. Ninety-three percent of respondents felt that the greatest benefit of QOL information is 'being able to treat the whole patient'. Forty one percent of respondents felt that length of survival is more important to patients than quality of life. However, only 7% of the respondents felt that the primary job of physicians is to save lives, and that QOL should not be a predominant concern for physicians. The inclusion of QOL in randomized trials was perceived as encouraging both patient and physician participation. The results of this survey are being used to further explore these critical issues. PMID- 9205969 TI - Predictors of parental emotional adjustment to childhood cancer. AB - The main objective of the present study was to determine which variables predict the emotional adjustment of parents of children with cancer. Therefore, parents' emotional adjustment, in terms of depression, anxiety, feelings of loneliness, helplessness, uncertainty and positive feelings, were predicted with three models. (1) With a child model (including age of the child, time since diagnosis, being in remission or having a relapse, and depression of the child); (2) with a control strategies model (including four distinguished control strategies of parents); and (3) with a child and control strategies model (including a combination of the aforementioned variables). The four control strategies of parents of children with cancer included: the reliance on predictive control (having positive expectations); vicarious control (attributing power to the medical setting); illusory control (relying on luck and wishful thinking); and interpretative control (gaining knowledge). A total of 84 mothers and 79 fathers, of 84 children with cancer with different survival perspectives (in remission or with a relapse) participated in the study, and were assessed about the use of control strategies and adjustment. Lack of positive expectations about the course of the illness was most strongly related to negative emotions for mothers and for fathers. For mothers having a child with a relapse, predicted feelings of helplessness and uncertainty, and reported feelings of depression of the child, proved to be related to the feelings of uncertainty of the fathers. The findings demonstrate that the use of secondary control strategies contribute significantly to the emotional adjustment of parents of children with cancer. PMID- 9205970 TI - Phase II study of psychotherapeutic intervention in advanced cancer. AB - The effect of psychosocial counseling on tumor progression was studied in 96 cancer patients, who were no longer amenable to regular medical treatment. Patients were offered 12 session of individual experiential-existential counseling, each sessions lasting 1.5 to 2 hours. In addition patients participated fortnightly in group counseling meetings. In five out of 35 evaluable patients, tumor growth became stationary during or immediately following therapy. In four patients this stationary period last 3-9 months, and in one patient 2 years. Natural Killer cell activity, self-reported loneliness, depression, purpose in life and locus of control showed no change from pre- to post intervention. PMID- 9205971 TI - Consultations for 'maladaptive denial of illness' in patients with cancer: psychiatric disorders that result in noncompliance. AB - Patients who present with late stages of cancer often have complicated medical and psychiatric problems which are labeled as 'maladaptive delay or denial.' In some of these patients, psychiatric problems have either contributed to the delay in medical presentation for care or have interfered with treatment of the late stage cancer. The authors review some of the factors that contributed to delay and noncompliance in a series of patients with cancer who were evaluated by the psychiatric consultation service of a university hospital. Specifically, psychoses and cognitive impairment played a major role in delay and noncompliance. The authors discuss recommendations for management of such patients, and suggest that clinicians often benefit from the assistance of the psychiatric consultant as part of the treatment team. Multiple resources and multiple types of intervention are needed in order to help such patients negotiate the clinical environment. PMID- 9205972 TI - Acute effects on neuropsychological function and quality of life by high-dose multiple daily fractionated radiotherapy for malignant astrocytomas: assessing the tolerability of a new radiotherapy regimen. AB - Cognitive and other quality of life measures were assessed in 29 patients with supratentorial malignant astrocytomas before and after high-dose (8000 cGy) multiple daily fractionated radiotherapy. Assessments were done immediately before and after radiotherapy. Patients completed a neuropsychological evaluation and the Functional Living Index: Cancer (FLIC). Spouses completed the Family Environment Scale and the Profile of Mood States. Cognitive abilities generally improved over the course of radiotherapy. Occasionally, deterioration of potential clinical importance was observed on functions associated with the tumour site. Quality of life as assessed by the FLIC was stable in most cases and improved in five, but deteriorated in three patients. Families showed slightly less Conflict and slightly more Cohesion than the norm; this was especially so when patients had greater cognitive deficit. Emotional state of spouses was variable, with increased fatigue or reduced activity most commonly reported, followed by depression and anxiety. Mostly this improved with time or remained stable, but two spouses reported worsening emotional state. Results are generally encouraging for tolerance of this radiotherapy protocol, although they demonstrate that limited adverse effects may occur in some cases. PMID- 9205973 TI - Medical outcomes in preterm infants. AB - Many survivors of the newborn intensive care units who were premature do very well; some, however, go on to have a variety of medical complications related, in part, to their prematurity. An overview of the medical outcomes of prematurity are discussed in the areas of respiratory disease (bronchopulmonary dysplasia), gastrointestinal disorders (short gut syndrome and gastroesophageal reflux), growth and nutrition problems, vision, and hearing outcomes. These complications can be managed on a regular or vigilant outpatient basis and, if exacerbated, may require hospital management. Concepts to assist in family counseling on expected long-term medical outcomes of prematurity are discussed. PMID- 9205974 TI - The role of relationship-based developmentally supportive newborn intensive care in strengthening outcome of preterm infants. AB - This article details the conceptual framework, clinical application, and efficacy of a relationship-based developmentally supportive approach to newborn intensive care referred to as NIDCAP (Newborn Individualized Developmental Care and Assessment Program). Outcomes of the approach are reported in regard to infant health and development, reduction of hospital costs, and family adaptation. The approach is guided by a neurodevelopmental framework for understanding preterm infants and depends on the capacities of professionals to collaborate with one another and with families in support of the infants' medical, developmental, and emotional well-being. The primary vehicle for clinical implementation is detailed behavioral observation with subsequent recommendations for individualized caregiving based on the infant's current functioning and apparent developmental goals. A series of essential components of developmentally oriented caregiving are described, including strategies for coordinated discharge planning, and linkage to community services. The voices of individual clinicians highlight the process of change from protocol-based to relationship-based care. PMID- 9205975 TI - Long-term perspective on premature infant outcome and contemporary intervention issues. AB - Despite improvements in survival rates for low birthweight (LBW) infants, the prevalence among survivors of major neurodevelopmental impairment seems relatively stable. Cerebral palsy, the most common major impairment, can usually be ruled out by 18 months corrected age. Minor impairments such as learning disabilities cannot be ruled out until much later. The efficacy of interventional services in this population was addressed by a national randomized trial. The intervention produced large treatment effects for heavier LBW infants and moderate effects for lighter infants. Five years later, modest residual effects were found for heavier LBW infants, but not for the lighter, suggesting that 0 to 3 services alone are not sufficient to prevent scholastic disadvantage in this population. PMID- 9205976 TI - Neuropsychological and functional outcomes of very low birth weight infants. AB - Advances in perinatal and neonatal management have resulted in a significant increase in the survival of fragile extremely low birth weight (ELBW) infants > 1,000 g at birth. The evaluation and reporting of the outcome of these infants aids in assessing the efficacy of interventions, provides data to aid in policy decisions, and provides critical information for parents and primary care providers. Comprehensive assessment of multiple domains including neurologic/neurosensory, developmental-cognitive, visual perceptual, speech/language, motor, functional skills for daily living, and Kindergarten readiness permit a total view of the child within the context of the family. Survival of VLBW infants < 800 g has steadily improved from 0% (1943 to 1945) to 49% to 70% (1994 to 1995). Rates of cerebral palsy, mental retardation, blindness, and deafness have remained stable in the 1980s and 1990s. There is evidence, however, that the percent of functional limitations may be increasing. A requirement for Special Education Resources among VLBW infants remains high at 44% to 56%. As increasing numbers of infants at the limits of viability survive, the medical community must remain vigilant in its surveillance and advocate both humanistically and scientifically for comprehensive strategies that optimize long term functional, academic, and family outcomes. PMID- 9205977 TI - Behavioral effects of prematurity. AB - This report reviews the evidence for an increased incidence of behavior and social problems in infants and children born prematurely. The contribution of biological and social factors to the development of behavior problems in this population is also examined. The available evidence indicates that preterms more often than full-terms exhibit negative temperament characteristics, symptoms of Attention-Deficit Hyperactivity Disorder, and lower levels of social competence. The risk for these problems appears to be limited to those infants with a birth weight of less than 1,500 g. Adverse social conditions also impact the expression of these problems. Preterms do not appear to be at as much risk for emotional or conduct problems or abnormal attachment to their mothers. Both the experience of a preterm birth and the characteristics of the infant can alter the perceptions and behavior of parents. Appropriate interventions should involve the child, the parents, and the school. PMID- 9205978 TI - School age outcome in low birth weight preterm infants. AB - Very low birth weight (VLBW) children at school age show variability in their outcome, compared with normal birth weight children, although many early physical and health differences are equalized by middle childhood. Studies of nonhandicapped VLBW children have found a higher rate of school retention and school problems in this population. Differences in intelligence have been reported, although these are often confounded by socioeconomic factors such as educational level of the parent. Few studies today of children born in the late 1970s and early 1980s have related school age outcome to central nervous system (CNS) status, yet for learning disabilities or other neuropsychological deficits, this may be highly relevant. Better understanding of medical risk factors, however, will not affect the decisive influence of social factors on their expression in the school age child. PMID- 9205979 TI - Parenting the prematurely born child: pathways of influence. AB - Recognizing the importance of parents in the lives of preterm infants, investigators and clinicians have increasingly focused on the needs of parents during the period when their infant is hospitalized in a neonatal intensive care unit and the impact of this experience on their subsequent parenting. The purpose of this report is to summarize research findings from over two decades of research, present a framework for understanding the various influences on parents of prematurely-born children, and suggest clinical interventions that are important in helping parents both in the hospital and after discharge. PMID- 9205980 TI - Chelation therapy for cardiovascular disease. Review and commentary. PMID- 9205981 TI - Antiplatelet therapy with glycoprotein IIb/IIIa receptor inhibitors and other novel agents. AB - Aspirin has stood the test of time over decades as the gold standard for relatively effective, safe, and inexpensive antiplatelet therapy. However, aspirin is only modestly effective in preventing arterial thrombosis in certain settings, and it is virtually ineffective in others (for example, preventing coronary restenosis after angioplasty). These observations have been the impetus for the development of more effective antiplatelet strategies, the rational basis of which is largely our understanding of normal platelet function. The most clinically effective platelet inhibitors yet developed produce broad inhibition of platelet function by blockade of the final common pathway of aggregation, in which fibrinogen binds to its platelet membrane receptor localized in the glycoprotein (Gp) IIb/ IIIa complex. The Gp IIb/IIIa complex is a member of the family of integrins, which are cell membrane receptors for adhesive proteins. The binding of fibrinogen to platelet Gp IIb/IIIa occurs via a specific amino acid sequence, arginine-glycine-aspartic acid. Effective antagonists of platelet Gp IIb/IIIa function have included monoclonal antibodies against Gp IIb/IIIa, peptide (peptidomimetic) antagonists of Gp IIb/IIIa, and nonpeptide (nonpeptide mimetic) antagonists of Gp IIb/IIIa. The major risk of any antiplatelet strategy is the potential for bleeding complications, since currently we do not understand the molecular distinction between protective hemostasis and pathologic thrombosis. PMID- 9205982 TI - Outcomes among pediatric heart transplant recipients. AB - Postoperative cytomegalovirus prophylaxis with cytomegalovirus immunoglobulin or ganciclovir has decreased the incidence of cytomegalovirus disease in cytomegalovirus-negative recipients of cytomegalovirus-positive donor organs. In adults, these drugs have also been used to treat recipients who developed symptomatic cytomegalovirus disease. This report describes outcomes of predominantly cytomegalovirus-negative pediatric cardiac transplant recipients of cytomegalovirus-positive donor organs who received cytomegalovirus immunoglobulin plus ganciclovir as cytomegalovirus prophylaxis, as well as results of this combination therapy when used to treat cytomegalovirus disease. We reviewed the records of children who received donor hearts at our institution between 1989 and 1994. Cytomegalovirus-negative patients who received cytomegalovirus-positive donor organs were given prophylaxis consisting of ganciclovir (5 mg/kg every 12 hours for 14 days, followed by maintenance dosage of 5 to 6 mg/kg every day for 14 days) plus 7 scheduled cytomegalovirus immunoglobulin infusions. Cytomegalovirus infection was documented by culture, polymerase chain reaction, and cytomegalovirus immunoglobulin M seroconversion of a 4-fold or greater rise in cytomegalovirus immunoglobulin G titers. After infection, patients were diagnosed with cytomegalovirus disease when they developed clinical symptoms. These episodes were treated with cytomegalovirus immunoglobulin infusions plus ganciclovir (5 mg/kg every 12 hours) until symptoms resolved. Of 40 cardiac transplant recipients, 10 cytomegalovirus-negative and 9 cytomegalovirus-positive patients received cytomegalovirus-positive donor organs. Five patients (3 of whom were seronegative and had received dual-agent prophylaxis) developed cytomegalovirus disease, which resolved with dual-agent therapy. During an average 15-month follow-up period, no significant morbidity or mortality was attributable to cytomegalovirus disease. Post-transplant dual-therapy cytomegalovirus prophylaxis appears to be as safe and effective in children as in adults, when our results are compared with the published results of studies in adults. Dual-agent treatment eradicated symptoms among patients who developed cytomegalovirus disease. This regimen may allow safer use of the cytomegalovirus positive donor pool for pediatric recipients. PMID- 9205983 TI - Right ventricular outflow obstruction with intact ventricular septum in adults. AB - Cardiothoracic surgeons whose practice is limited to adults rarely see patients with right ventricular outflow obstruction and an intact ventricular septum. Of more than 10,000 open-heart procedures performed at our institution from 1983 to 1993 (in patients 18 to 75 years old), only 5 procedures were for correction of this problem. Both the pulmonary valve and the subvalvular area were abnormal in these 5 patients, and 4 of the 5 had subvalvular stenosis. The gradient across the right ventricular outflow tract was measured by cardiac catheterization before repair in all patients and averaged 118 mmHg. Various surgical approaches were used for repair. In the 2 patients whose pressures were measured postoperatively, the gradients were 25 mmHg and 45 mmHg, respectively. There were no operative deaths. At follow-up (range, 2 months to 5 years after surgery), all patients were in New York Heart Association functional class I and all had murmurs. Those who underwent echocardiography were found to have minimal gradients across the right ventricular outflow tract. PMID- 9205985 TI - Use of the retrograde "pull-through" technique. AB - A 50-year-old man, who 9 months earlier had undergone emergency operation for acute type I aortic dissection, was readmitted to our hospital with the diagnosis of an enlarging aneurysm of the false lumen involving the transverse arch and the proximal third of the descending thoracic aorta, due principally to redissection at the distal suture line of the ascending aortic graft. Replacement of the aortic arch and proximal descending thoracic aorta was performed by using the retrograde "pull-through" technique with hypothermic circulatory arrest and retrograde cerebral perfusion. Although circulatory arrest lasted 110 minutes, the patient was extubated on the 2nd postoperative day and had no central or peripheral neurologic damage. Mild, transitory renal dysfunction was observed in the 1st postoperative week, and the patient was discharged on the 18th postoperative day. He is asymtomatic at 15 postoperative months. Deep hypothermia and retrograde cerebral perfusion proved effective despite prolonged circulatory arrest. The retrograde "pull-through" technique is an effective method of replacing the entire thoracic aorta and should probably be considered for single stage repair of acute type I aortic dissection with multiple intimal tears. PMID- 9205984 TI - Medical necessity for right heart catheterization. AB - Because there are no definitive guidelines for performing right heart catheterizations or controlled clinical trials demonstrating medical benefit, the value and necessity of performing routine right heart catheterizations for coronary artery disease have been questioned. This Texas Medical Foundation Health Care Quality Improvement Program project was designed to ensure medical necessity and proper documentation of right heart catheterization when performed as part of a bilateral procedure. Medicare claims data were used to identify Texas facilities where rates of bilateral catheterizations suggested that right heart catheterizations were being performed routinely. Five facilities were found to have rates of bilateral procedures exceeding 70%. Suggested guidelines for performing right heart catheterizations were prepared by the Texas Medical Association Committee on Cardiovascular Diseases. These guidelines, together with the facility's data on its rate of right heart catheterizations, were presented by the Texas Medical Foundation to the staff of each facility. They were asked to examine their individual facility's procedures for ensuring medical necessity and to develop and implement process improvement plans. Medicare claims data were analyzed to determine the rates of bilateral catheterizations before and after the plans were instituted. The statewide rate of bilateral procedures decreased from 27.2% to 21.3% (p < 0.005). Rate reductions for 4 facilities implementing improvement plans were statistically significant (p < 0.001): at the 1st facility, the rate decreased from 74.3% to 25.0%; at the 2nd, from 85.0% to 21.0%; at the 3rd, from 76.7% to 17.7%; and at the 4th facility, from 85.4% to 42.9%. The rate for the facility not implementing an improvement plan increased from 86.4% to 89.1%. Reductions in rates of bilateral procedures at the 4 facilities suggest that many procedures previously performed were routine and not medically indicated. Presentation of data and practice guidelines to facilities may have contributed to their ability to improve processes. PMID- 9205986 TI - New bipolar leads for the study of atrial arrhythmias. AB - The electrocardiography study of atrial arrhythmias has been hindered by the frequent occurrence of a "hidden". P wave in standard lead recordings. We present a modified method of surface electrocardiography that proved helpful in our prospective study of 26 consecutive patients in whom traditional surface recordings did not show the atrial mechanism. We recorded leads I, II, and III in their customary positions, except that we repositioned a pertinent limb electrode at a precordial site, in order to identify a P wave. The 4 alternative sites that we found most helpful were at the high and low parasternal positions, both to the right and to the left of the sternum. Confirmatory examples are submitted. PMID- 9205987 TI - Fracture embolization of a Duromedics mitral prosthesis. AB - The Duromedics bileaflet pyrolitic carbon mechanical prosthesis was introduced by Hemex in 1982 and subsequently acquired by Baxter. This communication documents a case of sudden leaflet fracture of a Duromedics mitral valve 48 months after implantation, which was managed successfully by replacement with a St. Jude Medical mechanical prosthesis. The patient presented in acute distress with paroxysmal atrial tachycardia and pulmonary edema. Transesophageal echocardiography was used to diagnose the leaflet fracture. The fracture had occurred transversely, with the fragments embolizing bilaterally to the iliofemoral arteries. These were removed at a subsequent operation. Cases of such fractures of the Duromedics prosthesis have been reported, with cavitation damage being the postulated mechanism. PMID- 9205988 TI - Carcinoid heart disease with severe hypoxia due to interatrial shunt through patent foramen ovale. AB - Carcinoid heart disease occurs in approximately half of patients who have carcinoid syndrome and is the leading cause of death among these patients. It is typically manifest as right-sided valvular lesions, usually tricuspid insufficiency and pulmonary valve stenosis. This case report describes the unusual presentation of a patient with carcinoid heart disease and a large right to-left shunt through a patent foramen ovale. PMID- 9205989 TI - Infection of a pacemaker by Brucella melitensis. AB - We report a case that, to the best of our knowledge, is the only published instance of infection of a pacemaker and its leads by Brucella melitensis. Furthermore, this case suggests that B. melitensis may be able to persist around pacemaker devices despite its having been eliminated from the rest of the body. The patient was a sheep shearer who had just undergone a 45-day course of antibiotic therapy for brucella and had been considered cured on the basis of negative blood cultures. PMID- 9205990 TI - Abdominal aortic aneurysm and congenital pelvic kidney. A rare association. AB - We report the case of a 65-year-old man who presented with an infrarenal aortic aneurysm in association with a congenital right pelvic kidney vascularized by 2 aortic arteries, 1 of which arose from the aneurysmal aorta and the other from the common right iliac artery. Successful surgery consisted of excising the aneurysmal aortic segment and replacing it with a Dacron tube graft, then implanting the upper renal artery (supplemented by a short segment of saphenous venous graft) in the Dacron prosthesis. We review 6 other cases of this rare pathologic association, found in our search of the literature, and discuss techniques of renal protection and (when necessary) reimplantation of the anomalous arteries. PMID- 9205991 TI - Repair of a ruptured sinus of Valsalva aneurysm. Associated with annuloaortic ectasia and coarctation of the aorta in a patient with Marfan syndrome. AB - We report the case of a 16-year-old boy with Marfan syndrome who presented in severe congestive heart failure secondary to rupture of an aneurysm of the sinus of Valsalva into the right atrium, a condition that was aggravated by coarctation of the aorta. The patient also had a large aneurysm of the ascending aorta with the characteristics of annuloaortic ectasia. The patient underwent successful surgical correction and is asymptomatic 3 years after the repair. PMID- 9205992 TI - Apical hypertrophic cardiomyopathy. PMID- 9205993 TI - Foreign body in the heart. PMID- 9205995 TI - Congenital isolation of the subclavian artery in adults. PMID- 9205994 TI - History of cardiac surgery. PMID- 9205996 TI - Malignant mesothelioma: current conundrums over risk estimates and whither electron microscopy for diagnosis? PMID- 9205997 TI - Analysis of asbestos fiber burden in lung tissue from mesothelioma patients. AB - Mesothelioma is a rare neoplasm that occurs most frequently in individuals with previous asbestos exposure. Differences for risk of development of asbestos related mesothelioma and lung cancer have been attributed to the various types of asbestos, as well as to the dimension of the inhaled fibers. In the present study, 55 individuals with the pathological diagnosis of mesothelioma were evaluated as to ferruginous body and fiber content in lung tissue. The procedures used in the analysis included tissue digestion and analysis of the collected material for ferruginous bodies by light microscopy and for uncoated fibers by analytical transmission electron microscopy. Forty-six of the samples had ferruginous body concentrations of over 1000/per gram dry weight of lung tissue. The majority of the cores of these ferruginous bodies were amosite. Likewise, the most common uncoated asbestos fiber in the tissue was amosite. Only a small percentage of each type of asbestos would have been visible by light microscopy or even potentially by electron microscopy if the magnification was not sufficient to detect those with thin (< 0.2 micron) diameters. The consistent finding in most of the cases was a considerable presence of asbestos, often of mixed types. PMID- 9205998 TI - Crystalloidal paraprotein deposits in the cornea: an ultrastructural study of two new cases with tubular crystalloids that contain IgG kappa light chains and IgG gamma heavy chains. AB - The fine structure and immunoprotein content of the crystalloids are described in two cases of paraproteinemic crystalloidal keretopathy, both of which had clinical features thought by the referring ophthalmologists to be those of atypical lattice-type corneal dystrophy (presumably because of lattice-like lines). Most keratocytes in one case were surrounded by a mantle of densely packed tubular crystalloids. Individual tubules were annular in cross section with mean dimensions as follows: overall diameter, 29.32 nm (SD 1.26); internal diameter (core), 8.53 nm (SD 1.12); wall thickness, 10.39 nm (SD 0.85) (n = 10). Crystalloids were extracellular and found only in the corneal stroma, with none in Bowman's layer or Descemet's membrane. In the second case, the tubules had a similar distribution but formed geometric arrays with no clear relationship to, or envelopment of the keratocytes. The tubules were thin-walled, with mean dimensions as follows: overall diameter, 26.12 nm (SD 1.12); internal diameter (core), 15.46 nm (SD 1.12); wall thickness, 5.33 nm (SD 0) (n = 10). In both cases the tubules were kappa-light chain- and gamma-chain-positive. Laboratory investigations revealed the presence of two IgM-kappa paraproteins and an IgG kappa paraprotein in the serum of the first patient. The second patient had an IgG-kappa paraproteinemia and bone marrow changes consistent with low-grade non Hodgkin's lymphoma. These cases emphasize and extend the morphological range of corneal IgG crystalloids; the second case also demonstrates that corneal IgG crystalloids may be an early indicator of un underlying immunoproliferative disease. PMID- 9205999 TI - Role of electron microscopy in the diagnosis of metabolic storage diseases affecting the nervous system of children. AB - Metabolic storage diseases are among the most challenging diagnostic problems faced by clinicians and pathologists. There is considerable variation in the diagnostic approach to these diseases between different institutions and between different diagnosticians. Much of this variation arises from differences in the availability of and physician confidence in the diagnostic modalities employed to characterize these disorders. Recent advances in the biochemistry and molecular genetics of these diseases have produced some skepticism about the continued relevance of traditional morphologic techniques, including electron microscopy, in their diagnosis. It is the opinion of the authors that this concern is premature and that electron microscopy continues to play a vital role, particularly in the diagnosis of those entities that challenge the classic definitions of lysosomal storage diseases. The authors present a series of cases illustrating different situations where electron microscopy can provide timely, cost-effective, and accurate information in the workup of such diseases. PMID- 9206000 TI - Early ultrastructural changes in the central nervous system in Fukuyama congenital muscular dystrophy. AB - Electron microscopy of the central nervous system surface structure is described in two fetuses with Fukuyama congenital muscular dystrophy (FCMD). In addition to relatively large surface defects, many minute defects less than several micrometers in size associated with protrusion of glial cytoplasm were observed in the cerebrum. These findings were considered to represent early changes prior to cortical dysplasia. The basement membrane adjacent to the defects showed amorphous, wavy, or whorled configurations, and gradually disappeared. The glial cytoplasmic membrane seemed to be relatively well preserved in some areas where the basement membrane disappeared. On the other hand, both the basement membrane and cytoplasmic membrane became indistinct irregularly in areas without defects, including the spinal cord; similar lesions were found in the skeletal muscle. These observations confirm previous observations concerning defects of the pial glial barrier of the brain surface, and may suggest the involvement of abnormal basement membrane or related structures, or both, in the genesis of the brain lesions of FCMD. PMID- 9206001 TI - Primary malignant rhabdoid tumor of the central nervous system. AB - Since the initial description of malignant rhabdoid tumor (MRT) of the kidney by Beckwith in 1978, MRTs have been established as a distinct clinicopathologic entity lacking nephrogenic and myogenic differentiation. MRTs are highly aggressive neoplasms with characteristic histopathologic, immunocytochemical, and ultrastructural features. Many reports have appeared documenting primary extrarenal rhabdoid tumors (ERRTs) occurring at diverse sites, including infratentorial and supratentorial compartments of the central nervous system (CNS). The authors report 2 cases of primary CNS-MRT in young male children (6.5 and 7 years of age) and review the literature on CNS-MRTs. Neuroimaging studies showed an inhomogeneous parasagittal mass in the left anterior parietal region involving the motor strip and attached to the lateral aspect of the superior sagittal sinus in one case, and a right parietal parasagittal tumor with a cystic component in the other case. Metastatic workup, including abdominal CT, was negative in both cases. Histologic examination of the resected tumors showed irregular clusters and nests of cells with variable desmoplasia in both cases. Large areas of tumor necrosis and apoptotic tumor cells were present. Prominent eosinophilic cytoplasmic inclusions and eccentric, indented nuclei with conspicuous nucleoli characterized many of the tumor cells. Diffuse strong vimentin reactivity and focal strong reaction for epithelial membrane antigen (EMA) were demonstrated. Cytogenetic analyses reported a normal male karyotype in one case and an abnormal male karyotype with loss of both normal copies of chromosome 22 and gain of one structurally rearranged chromosome 22 in the other case. Ultrastructural examination displayed tumor cells with avoid to indented nuclei, marginated chromatin, and prominent nucleoli. Intercellular junctions were not found. Masses of cytoplasmic intermediate filaments in a characteristic whorled configuration were present. CNS-MRTs are consistently vimentin positive (100%) and usually EMA positive (90%). Glial fibrillary acidic protein, neuron specific enolase, and S-100 protein are variably expressed. Markers for myogenous differentiation are invariably absent. Ultrastructural characteristics include aggregates of intermediate filaments. Monosomy 22 occurs in some CNS rhabdoid tumors, while most renal rhabdoid tumors are cytogenetically normal with only isolated cases having del(13q), del(11p), del(22)(q11), and unbalanced reciprocal translocation involving chromosomes 8 and 22. The prognosis for CNS rhabdoid tumors is dismal and almost two-thirds of patients are dead of disease shortly after diagnosis; one-third have been reported to be alive with disease, but have been followed for only short periods; and a single patient is reported to be free of disease at 5 years. PMID- 9206002 TI - Central nervous system atypical teratoid/rhabdoid tumors of infancy and childhood. AB - In 1987, a distinctive brain tumor arising in young children was first described. This tumor contained neuroepithelial, peripheral epithelial, and mesenchymal elements, but lacked divergent tissue differentiation characteristic of malignant teratomas. It was originally designated as atypical teratoid tumor, but because of the prominent rhabdoid component, the tumor designation was modified to atypical teratoid/rhabdoid tumors (AT/RT) of infancy and childhood. AT/RTs occur most commonly in infants under 2 years of age, often have central nervous system (CNS) dissemination, do not respond to therapy, and typically are fatal within 1 year. Most are located in the cerebellum (65%), but they may arise at any CNS site. Histologically, various patterns can be present within the same tumor, but they all have a population of rhabdoid cells, and 70% contain fields typical of a primitive neuroectodermal tumor (PNET/medulloblastoma). Less frequently, malignant mesenchymal tissue and/or an epithelial component are found. Necrosis and brisk mitotic activity are common. The immunocytochemical profile is complex, but germ cell markers are consistently negative. Ultrastructural features vary and depend on the site sampled, but whorled bundles of cytoplasmic intermediate filaments are a distinctive finding in cells of the rhabdoid component. The authors report 4 AT/RTs (2 males, 2 females, age range 6 months to 4 1/2 years, 3 cerebellar, 1 cerebral). All cases showed a variety of histologic patterns with necrosis. Typical rhabdoid cells, PNET areas, undifferentiated bland large cell regions, dense connective tissue, and solid clusters of epithelial cells were present. Immunocytochemistry showed strong vimentin reactivity, whereas epithelial membrane antigen, cytokeratin, glial fibrillary acidic protein, S-100 protein, desmin, and smooth muscle actin were present to a lesser extent in most cases. Germ cell markers were negative. Ultrastructurally, many cells contained aggregates of cytoplasmic intermediate filaments, and some cells had a basal lamina on one aspect. Cells with interdigitating cytoplasmic borders were seen and rare cells had microtubules. Cytogenetic studies were normal in 2 cases. Follow-up has shown that 3 children have died of disease (< 1 year after diagnosis) and 1 child is alive with disease (18 months after diagnosis). Separation of AT/RT from PNET based on histopathologic and biologic evaluation is important, because AT/RTs are aggressive tumors with a dismal prognosis and currently there is no effective treatment. Neither clinical signs and symptoms nor radiologic features will distinguish AT/RTs from PNETs. PMID- 9206003 TI - Ultrastructural study of the nuclei of normal, dysplastic, and carcinomatous epithelial cells of the human cervix uteri. AB - The nuclei of epithelial cells of the uterine cervix of normal women and of patients with various degrees of dysplasia, carcinoma in situ, and invasive carcinoma were studied by means of electron microscopy. Nuclear ribonucleoprotein components and chromatin were contrasted using preferential methods for RNA and DNA. Changes in the distribution of the extranucleolar ribonucleoprotein containing structures were found, ranging from low-grade dysplastic lesions to invasive carcinoma. Compared with normal epithelial cells, dysplastic and neoplastic cells possess more nuclear bodies, as well as deep invaginations of the nuclear envelope and lobulations. Morphometric parameters estimated were nuclear volume, numerical density of perichromatin granules (PCG), and fraction of nuclear volume occupied by compact chromatin. The pattern of values of these parameters in the cell layers of normal cervical epithelium was disrupted in all the lesions. These data suggest that the processes studied induce early alterations in transcription and processing and/or exportation of mRNA to the cytoplasm. Two populations of cells were found in invasive carcinomas, one with large nuclei, sparse compact chromatin, and few PCG, and the other with small nuclei, abundant compact chromatin, and numerous PCG. Their morphologic features indicate that the former population is composed of relatively undifferentiated cells, while the letter is made up of well-differentiated cells which could be neoplastic or entrapped normal cells. PMID- 9206004 TI - Ultrastructural characteristics of central neurocytoma in cell culture. AB - The study was performed to determine the ultrastructural characteristics of central neurocytoma and its features in primary cell culture. Fresh tissue from three tumors was mechanically and enzymatically dissociated into individual cells, which were cultured onto poly-L-lysine precoated Aclar coverslips in the media. The tumor cells attached to the surface of the coverslips within 12 to 24 h and delicate cytoplasmic processes developed within 2 to 3 days. Electron microscopic examination of the cultured tumor cells and the tumor tissue supported neuronal origin. Combined tissue culture and electron microscopic study provides a rapid, reliable, and reproducible means for the diagnosis of central neurocytoma. PMID- 9206005 TI - Polycyclic aromatic hydrocarbon bioremediation design. AB - Many polycyclic aromatic hydrocarbons (PAHs) are known to be mutagenic or carcinogenic, and their contamination in soil and aquifer is of great environmental concern. Limited numbers of microorganisms including mycobacteria, Sphingomonas and white rot fungi were found to be capable of degrading PAHs with four or more fused aromatic rings. In white rot fungi, lignin peroxidases are believed to be involved in the degradation of PAHs. In addition to these enzymes, P450 monooxygenases in some fungi were implicated in the degradation of PAHs. The stimulation of PAH biodegradation by the addition of surfactants was observed with some of these microorganisms although the agents were inhibitory on biodegradation with some other microorganisms. Mathematical models were constructed to explain the effect of surfactants on biodegradation. Further studies should be carried out to select the best microorganisms and surfactants for applications to PAH bioremediation. PMID- 9206006 TI - Engineering cytochrome P450s for bioremediation. AB - Building on the vast knowledge of active site structure and catalytic mechanisms of the P450 monooxygenase systems, significant efforts to utilize the rational design of engineered P450s are emerging as an approach to solve the problems of bioremediation. P450 enzymes are being designed to alter substrate specificities and catalytic efficiency in predefined ways. In addition, random mutagenesis and in vitro evolution are being considered as exciting methods for generating mutant P450s with increased bioremediation abilities. PMID- 9206008 TI - Bioremediation of metal contamination. AB - Recent studies have demonstrated that microbes might be used to remediate metal contamination by removing metals from contaminated water or waste streams, sequestering metals in soils and sediments or solubilizing metals to aid in their extraction. This is primarily accomplished either by biosorption of metals or enzymatically catalyzed changes in the metal redox state. Bioremediation of metals is still primarily a research problem with little large-scale application of this technology. PMID- 9206007 TI - Phytoremediation of soil metals. AB - The phytoremediation of metal-contaminated soils offers a low-cost method for soil remediation and some extracted metals may be recycled for value. Both the phytoextraction of metals and the phytovolatilization of Se or Hg by plants offer great promise for commercial development. Natural metal hyperaccumulator phenotype is much more important than high-yield ability when using plants to remove metals from contaminated soils. The hypertolerance of metals is the key plant characteristic required for hyperaccumulation; vacuolar compartmentalization appears to be the source of hypertolerance of natural hyperaccumulator plants. Alternatively, soil Pb and Cr6+ may be inactivated in the soil by plants and soil amendments (phytostabilization). Little molecular understanding of plant activities critical to phytoremediation has been achieved, but recent progress in characterizing Fe, Cd and Zn uptake by Arabidopsis and yeast mutants indicates strategies for developing transgenic improved phytoremediation cultivars for commercial use. PMID- 9206009 TI - Anaerobic dehalogenases. AB - Several anaerobic bacteria are able to reductively dehalogenate chlorinated hydrocarbons and to couple this reaction to the synthesis of ATP via a chemiosmotic mechanism (dehalorespiration). A few reductive dehalogenases have recently been purified and characterized. Preliminary investigations have been performed to elucidate the mechanism of dehalorespiration. PMID- 9206010 TI - Strategies for the aerobic co-metabolism of chlorinated solvents. AB - Recent field and laboratory studies have evaluated the potential for aerobic co metabolism of chlorinated solvents. Different co-metabolic substrates and different methods of application have been tried, including growing indigenous microbes in situ, and injecting into the soil subsurface strains grown in subsurface reactors for their co-metabolic potential. There is much potential for using co-metabolism for treating a broad range of chlorinated aliphatic hydrocarbons. Recirculation wells have potential for adding soluble co-metabolic substrates (i.e. phenol and toluene) into contaminated aquifers, while direct addition of gaseous substrates (i.e. methane and propane) into aquifers also holds promise. Aromatic substrates (phenol and toluene) are best used for treatment of chlorinated ethenes, whereas gaseous co-metabolic substrate (methane and propane) are better suited for the treatment of chlorinated methanes and ethanes. Many factors can enhance co-metabolic transformations, such as nutrients and available energy sources. PMID- 9206011 TI - Environmental bioadhesion: themes and applications. AB - Many marine organisms attach to underwater surfaces using protein adhesives. These are basic proteins with high levels of the amino acid 3,4 dihydroxyphenylalanine and an extended flexible conformation. The hydroxylation of tyrosine residues plays a key role in the chemisorption of these polymers to surfaces and in the setting of the adhesive. These unique proteins are attracting biotechnological attention for application in industry and medicine. Recent development on the immobilization of antigens and antibodies, enzymes, cells and tissues, illustrate the great potential use of these adhesives for diagnostics and medicine. The use of these adhesive proteins as anticorrosive coats for metal also suggests important applications for industry. PMID- 9206012 TI - Bioemulsans: microbial polymeric emulsifiers. AB - Bioemulsans are amphipathic proteins and/or polysaccharides that stabilize oil-in water emulsions. Bioemulsans are produced by a wide diversity of microorganisms and have potential applications in the food, paper, paint, bioremediation, agriculture, detergent and cosmetics industries. The production of the RAG-1 emulsan has been studied in batch-fed fermentors via self-cycling fermentation and with immobilized cells using a Celite support matrix. Bioemulsans have several advantages over lower molecular weight emulsifiers presently used in industry. The last few years have seen a number of new bioemulsans described with commercial applications. PMID- 9206013 TI - Prospects for altering host range for baculovirus bioinsecticides. AB - Advances in the understanding of baculovirus replication and the identification of genes that affect host range set the stage for constructing recombinant baculoviruses for specific past insects. The modification of baculovirus host specificity has recently been achieved by inserting or deleting genes that affect virus replication or cellular defenses. PMID- 9206014 TI - The ecology and biosafety of baculoviruses. AB - Advances in the use of molecular techniques-particularly for virus identification, the investigation of latency and the infection process, plus the development of a theoretical framework containing a higher degree of biological realism-have pushed baculovirus ecology forward in the past few years. This has created a scenario in which many hitherto intractable questions about the behaviour of natural and genetically modified baculoviruses can now be addressed. PMID- 9206015 TI - Understanding and advancing wastewater treatment. AB - In bioreactors used for the purification of wastewater, microorganisms are active in biofilms or aggregates. Insight into the factors that determine the structure and function of aggregated biomass is increasing steadily. Besides conventional techniques, modem molecular techniques are used increasingly to get a better understanding of the complex microbial communities in wastewater treatment systems. In recent years, the combined use of these techniques has led to a good insight into the population dynamics of different types of microbes in bioreactors. PMID- 9206016 TI - Biological waste air treatment in biofilters. AB - Recent studies in the area of biological waste air treatment in biofilters have addressed fundamental key issues such as microbial dynamics, microscopical characterization of the process culture and oxygen and nutrient limitations. The results from these studies have provided a deeper insight into the overall biofiltration process. In the coming years, such advances should allow for the design of better reactor controls and the improvement of pollutant removal in gas phase bioreactors. PMID- 9206017 TI - Biodiversity: are microbial species threatened? PMID- 9206018 TI - Regulatory affairs biopharmaceuticals regulation--progress and challenges. PMID- 9206020 TI - Vaccines: current regulatory issues. PMID- 9206019 TI - Potential influence of international harmonization of pharmaceutical regulations on biopharmaceutical development. PMID- 9206021 TI - Gene therapy: regulatory issues and international approaches to regulation. PMID- 9206023 TI - Environmental biotechnology. PMID- 9206022 TI - FDA regulatory reform. PMID- 9206024 TI - Systematic nonlinearities in the memory representation of time. AB - The representation of time was investigated by testing rats with intervals that changed by 2 s across trials. In Experiment 1, 2 ranges (20-150 s, 30-160 s; n = 10 rats per group) were examined. The times at which response bursts occurred (start time) were approximately proportional to interval durations. However, systematic departures from linearity were observed. Nonlinearities were related to the absolute duration of intervals, rather than to durations relative to the range. In Experiment 2, 660-s trials were inserted into the sequence of intervals (10-140 s, n = 20). Start and end times of response bursts were approximately proportional to intervals, but nonlinearities in start and end times were correlated, indicating that the source of nonlinearity was in the memory representation of time rather than in a decision process. These results indicate that the representation of time is nonlinearly related to physical time. PMID- 9206025 TI - Additivity of the effects of retention interval and context change on latent inhibition: toward resolution of the context forgetting paradox. AB - Three experiments with rats examined retention interval and context switch effects factorially in the latent inhibition paradigm. In Experiment 1, a 28-day retention interval abolished a context switch effect on latent inhibition. In Experiment 2, re-exposure to the contexts before conditioning re-established the context switch effect at the 28-day interval. In this case, the retention interval and context switch effects were additive: Latent inhibition was weakest when the retention interval and context switch were combined. Experiment 3 replicated the context switch effect at the 28-day interval. The results suggest that context switch and retention interval effects may be based on the same process. Context switch effects may weaken over time because physical contexts are embedded in superordinate temporal contexts; animals fail to retrieve physical context when the temporal context changes. This view helps resolve a paradox that has been noted for contextual change theories of forgetting. PMID- 9206026 TI - Converging evidence for one-trial context fear conditioning with an immediate shock: importance of shock potency. AB - In a sample of 208 Holtzman-descended albino rats, we found evidence with 4 measures of conditioning (freezing, defecation, side crossing, and nose crossing) that a single 2-s, 1.0-mA immediate shock could condition fear to a context (Experiments 1, 2, and 4). When we reduced the shock intensity to 0.5 mA, we obtained a complete immediate-shock conditioning deficit according to all measures in Experiment 3 and to all but the defecation measure in Experiment 4. Results suggest two conclusions: (a) Differences in shock potency between laboratories may help explain discrepant findings about whether immediate shock supports contextual conditioning; (b) theories of contextual conditioning need a mechanism that permits that conditioning to result from immediate shock. PMID- 9206027 TI - Judgements of ordinality and summation of number symbols by squirrel monkeys (Saimiri sciureus). AB - In Experiment 1, 2 squirrel monkeys (Saimiri sciureus) were given choices between all possible pairs of the arabic numbers 0, 1, 3, 5, 7, and 9, with choice of any number yielding that number of pieces of peanut as a reward. Both monkeys learned to choose the larger number in all pairings and learned to choose the largest number within a set of 4 numbers. In Experiments 2-4, the monkeys were tested on problems in which they chose between pairs of stimuli containing 2 numbers versus 2 numbers, 1 number versus 2 numbers, and 3 numbers versus 3 numbers. Both monkeys showed a significant tendency to choose the stimulus that contained the largest sum. Various tests indicated that this effect could not be explained by choice of the stimulus with the largest single number, by avoidance of the stimulus with the smallest single number, or by experimenter cuing. PMID- 9206028 TI - Interaction between piloting and beacon homing by rats in a swimming pool. AB - In three experiments, rats in a swimming pool were trained to find a submerged platform with a beacon attached to it. For some rats this beacon unambiguously identified the location of the platform; for others the beacon was made ambiguous by placement of an identical beacon in a different part of the pool. Test trials, in the absence of the platform and the beacons, revealed more persistent searching near the original location of the platform if the beacon attached to the platform had been ambiguous. These results show that learning about the location of the platform, with regard to cues that lie beyond the pool, is influenced by the extent to which an animal can find the platform by relying on other cues. The final experiment shows that this interaction between cues is influenced by an animal's prior experience. PMID- 9206029 TI - Models of ratio schedule performance. AB - Predictions of P. R. Killeen's (1994) mathematical principles of reinforcement were tested for responding on ratio reinforcement schedules. The type of response key, the number of sessions per condition, and first vs. second half of a session had negligible effects on responding. Longer reinforcer durations and larger grain types engendered more responding, affecting primarily the parameter alpha (specific activation). Key pecking was faster than treadle pressing, affecting primarily the parameter delta (response time). Longer intertrial intervals led to higher overall response rates and shorter postreinforcement pauses and higher running rates, and ruled out some competing explanations. The treadle data required a distinction between the energetic requirements and rate-limiting properties of extended responses. The theory was extended to predict pause durations and run rates on ratio schedules. PMID- 9206030 TI - Frog PNKT-4B cells express specific extracellular matrix-degrading enzymes and cytokines correlated with an invasive phenotype. AB - A temperature-dependent metastatic phenotype reported for a frog cell line, PNKT 4B, provided a means for studying potential mediators of cell-matrix interaction involved in metastatic invasion. Zymography revealed that these cells secreted enzyme species with properties and characteristics of mammalian metalloproteinases: collagenase, stromelysin, gelatinase A, and gelatinase B. These enzymes were produced by PNKT-4B cultures maintained at both invasive permissive (28 degrees C), and invasion-restrictive (20 degrees C) temperatures. However, under the invasive-permissive culture condition cells produced more of the putative gelatinase B and A enzymes. In addition, an activated form of gelatinase A was produced only in invasion-permissive cultures. DNA synthesis bioassays (Mv1Lu cell line and mouse thymocytes) to detect growth promoting and/or inhibitory cytokines, revealed that PNKT-4B cultures kept at 28 degrees C released significantly higher levels of stimulatory (interleukin-1-like) and latent inhibitory (transforming growth factor-beta-like) substances into the medium compared to 20 degrees C cultures. Pre-absorption of media samples with heparin-sepharose indicated a second stimulatory cytokine as well. A corneal fibroblast bioassay that tests for mediators of collagenase synthesis, detected a stimulatory substance whose activity was greatly reduced in the presence of interleukin-1 receptor antagonist protein. Collagenase stimulatory activity present in 28 degrees C culture medium was significantly higher than equal samples from 20 degrees C cultures. These studies provide a molecular correlation between release of cytokines with properties of the metastatic phenotype seen in vivo. They further provide some of the first characterizations of frog MMPs and cytokines, which are likely to be involved in other tissue remodeling events. PMID- 9206031 TI - Effects of estrogenic hormones on early development of Xenopus laevis. AB - Many chemicals released into the environment have estrogenic activity and can disrupt animal development and the function of endocrine systems. In order to study the effects of estrogens on aquatic animals, we examined the effects of certain estrogens on early development in Xenopus laevis. X. laevis embryos were kept in water containing 10(-10), 10(-9), 10(-7), 10(-6), and 10(-5) M 17 beta estradiol (E2); 17 alpha-estradiol; diethylstilbestrol (DES); 10(-5) M progesterone (P); or dihydrotestosterone (DHT) beginning at developmental stage 3. Survival rates of the embryos developed in water containing 10(-10)-10(-6) M E2 or DES, all concentrations of 17 alpha-estradiol, and 10(-5) M P or DHT, which were over 70% after stage 48, whereas the rates of the embryos treated with 10( 5) M E2 and DES decreased remarkably after stage 27 and all embryos were dead by stages 42 and 32, respectively. Embryos treated with 10(-5) M E2 showed malformations of the head and abdomen and suppressed organogenesis, including crooked vertebrae at stage 38; the head was smaller and the abdomen was larger than in the controls. Similar effects were observed in embryos developed in 10( 5) M DES but not in 10(-5) M 17 alpha-estradiol, P, or DHT. After 10(-5) M E2 treatment, abnormalities were induced only when the treatment was started before stage 39. However, on day 30 after fertilization, the stage of the embryos treated with 10(-6) M E2 was more progressed than that of the controls. Estrogen receptor (ER 4) mRNA was examined in eggs, embryos, and adult female liver by reverse-transcription polymerase chain reaction. ER4 mRNA was expressed in adult liver, unfertilized and fertilized eggs, and embryos, but ER3 mRNA was not expressed. ER4 mRNA in 10(-6) and 10(-5) M E2-treated embryos showed different expression patterns, which may result from the diverse developmental effects of E2. The present results demonstrate that 10(-5) M E2 and DES induced embryo death and malformations and that ER may be involved in the induction of various developmental defects in X. laevis embryos. PMID- 9206032 TI - Inhibitory role of prolactin in the downregulation of testicular follicle stimulating hormone receptors in mice. AB - Previous studies have shown that follicle-stimulating hormone (FSH) reduced its own receptors (downregulation) in the testis of adult mice. In the present study, we further examined the effect of prolactin (PRL) on the downregulation of testicular FSH receptors using hypophysectomized adult mice. FSH binding per testis increased after hypophysectomy, reaching a peak 5-10 days after the operation, and decreased thereafter. Hypophysectomized mice were given s.c. injections of 100 micrograms ovine PRL (NIADDK-oPRL) and/or 2 micrograms ovine FSH (NIADDK-oFSH) twice daily for 5 or 10 days beginning from 5 or 10 days after hypophysectomy. The 5-day treatment with a combination of oPRL and oFSH maintained the level of testicular FSH binding that was decreased by the treatment with oFSH alone, while the 5-day treatment with oPRL alone did not influence the testicular FSH binding. The change in the FSH binding per Sertoli cell was comparable to that in the binding per testis. A similar result was obtained by the 10-day treatment. Therefore, PRL was suggested to have an inhibitory role in the downregulation of testicular FSH receptors by FSH in adult mice. PMID- 9206033 TI - Full-term development after transfer of nuclei from 4-cell and compacted morula stage embryos to enucleated oocytes in the mouse. AB - The developmental ability of enucleated mouse oocytes reconstituted between the nucleus from 4-cell mouse embryos at different cell stages and recipient cytoplasms at different conditions, and the developmental ability of oocytes receiving nuclei from compacted morulae were examined. The highest development was observed with the nucleus at the G1 stage fused with cytoplasm at the M stage. Although the enucleated oocytes receiving a nucleus from 4-cell embryos did not develop after transfer to a recipient female, young were obtained after renuclear transfer to enucleated fertilized eggs. Live mice were also obtained from the nucleus of compacted morula. PMID- 9206034 TI - [Reversion of malignant phenotypes of human lung squamous carcinoma cells by ornithine decarboxylase antisense RNA]. AB - Abnormally elevated activity of ornithine decarboxylase (ODC), with subsequent polyamine accumulation are intimately associated with the genesis, development and metastasis of cancer. Therefore, ODC antisense RNA was used to transfect human lung squmous carcinoma cell line LTEP-78. Compared with the parental cells, growth of the antisense transfected LTEP-78 cells arrested in G0/G1 phase and colony formation in soft agar and tumorigenicity in nude mice were significantly reduced. Nucleic acid hybridization demonstrated the expression of ODC antisense RNA and the content of ODC mRNA was markedly reduced. The results suggest that the reversion of malignant phenotypes of human lung squamous carcinoma cells is associated with the control of polyamine biosynthesis. PMID- 9206035 TI - [Expression of transforming growth factor alpha and p53 in non-small cell lung cancer by immunohistochemical study]. AB - Primary non-small cell lung cancer samples were examined for the expression of p53, transforming growth factor alpha (TGF-alpha) and its receptor EGF-R by immunohistochemistry. Accumulation of p53 protein was found in 15 out of 24 carcinomas. Meanwhile, the results of TGF-alpha and p53 expression in 21 carcinomas showed that p53 protein was not detected in 2 out of 2 TGF-alpha negative and 4 out of 5 TGF-alpha low expression cases. However, there was no relationship found between EGF-R and p53 expression in these cases. The results suggest that p53 may have some effects on TGF-alpha expression. PMID- 9206036 TI - [A preliminary study of the relation between topoisomerase I and translocation (15;17)]. AB - The chromosomal translocation (15;17) which produced the PML-RAR alpha fusion gene had been found in acute promyelocytic leukemia (APL) cells. It was identified that the RAR alpha gene on chromosome 17 and PML gene on chromosome 15 were involved in this translocation. For study the molecular mechnisms of t(15:17) in APL, we cloned and sequenced the junctional region of the chromosomal reciprocal translocation in one APL patient. Furthermore, we compared 21 junctional sequences which had been reported with the consensus sequence of the DNA-topoisomerase I-binding sites. Thus, we proposed a hypothesis that topoisomerase I may play certain role in t(15;17) illegitimate recombination. PMID- 9206037 TI - [Experimental study on in vivo hematopoietic regulation of IL-6 gene-transfected tumor vaccine]. AB - In the present study, we investigated the effects on hematopoiesis of inactivated vaccines prepared from mouse erythroid leukemia cells (FBL-3) transfected with IL 6 gene. After treatment with IL-6 gene-transfected FBL-3 cells, the platelet count in mice started to increase at day 4, reached its maximum at day 10 and lasted for 25 days. The neutrophil count also increased significantly, but WBC count remained unchanged. The CFU-GM and CFU-MK were elevated to some extent in bone marrow and spleen. These results indicated that in addition to augmentation of antitumor immunity, IL-6 gene-transfected tumor vaccine could promote hematopoietic functions in bone marrow and spleen, and elevate platelet and neutrophil counts in peripheral blood. This approach to gene therapy may be applicable in leukemia treatment, especially for thrombocytopenia related to leukemia or chemotherapy. PMID- 9206038 TI - [Acceleration of hematopoietic reconstitution in mice transplanted with syngeneic bone marrow cells by IL-6 gene therapy]. AB - In the present study, the potential of fibroblast-mediated IL-6 gene therapy for accelerating hematopoietic reconstitution after syngeneic bone marrow transplantation (BMT) in BALB/c mice subjected to total body irradiation at a dose of 7 Gy was investigated. We observed that not only recovery of platelets, WBCs, CFU-GM or CFU-MK in bone marrow in mice treated with IL-6 gene therapy in combination with transplantation of 10(7) syngeneic bone marrow cells was faster than that in mice treated with IL-6 gene therapy in combination with transplantation of 10(6) or 10(5) syngeneic bone marrow cells, but also the number of CFU-S or survival rate in mice treated with IL-6 gene therapy in combination with transplantation of 10(7) syngeneic bone marrow cells was significantly higher. Nevertheless, the number of platelets, WBCs, CFU-GM or CFU MK in bone marrow, CFU-S and survival rate in mice treated with IL-6 gene therapy in combination with transplantation of syngeneic bone marrow cells were elevated more significantly than that in mice transplanted with syngeneic bone marrow cells alone. These data demonstrated that IL-6 gene therapy could markedly augment hematopoietic reconstitution after syngeneic bone marrow transplantation and the more bone marrow cells transplanted, the better was the effect. PMID- 9206039 TI - [Down-regulation of p53 expression of white blood cells in patients with myelogenous leukemia]. AB - By using RT/PCR technique the p53 phenotypes of normal white blood cells (WBC), leukemic cell lines and WBC from myelogenous leukemia patients were performed. The results showed that the p53 mRNA in normal bone marrow WBC was moderately expressed (p53 mRNA/beta-actin mRNA = 0.871 +/- 0.032) while it was not detectable in HL60 and K56z cell lines. The level of p53 mRNA in bone marrow WBC from 21 leukemia patients was significantly down-regulated (p53 mRNA/beta-actin mRNA = 0.433 +/- 0.181, P < 0.01). PMID- 9206040 TI - [A human bone marrow cell line (B3HM) inducing leukemia in mouse]. AB - Human bone marrow mononuclear cells were cultured on a layer of stromal cells derived from bone marrow of a patient with esophageal cancer. After five-week cultivation, the mononuclear cells could grow independently of stromal cells in liquid culture and had been passaged over 50 generations with a doubling time of 48 hours. The cell line thus established is called B3HM. The growth of B3HM appeared anchorage independent in semi-solid culture and produced 423 +/- 87.7 colonies per 5 X 10(4) cells. Immunofluorescence analysis showed that the cells were CD19+, CD20+, CD13+, CD15+, with both B cell and granulocyte surface markers. The cells or cell homogenate could induce leukemia when inoculated in nude mice and "615" inbred mice. The induced mouse leukemia was transplantable and L3T4 positive in more than 80% of the leukemic cells. The pathogenesis of the induced mouse leukemia is worthy of further investigation. PMID- 9206041 TI - [Highly metastatic model of human hepatocellular carcinoma established in nude mice using orthotopic organ selection of metastatic variant from patient specimens]. AB - We succeeded in establishing a highly metastatic model of human hepatocellular carcinoma (HCC) in athymic nude mice (LCI-D20). The metastatic variant was obtained by using orthotopic implantation of histologically intact tumor tissue selected from 30 fresh surgical specimens. It exhibited various features seen in clinical liver cancer patients: local growth, regional invasion, spontaneous intrahepatic, lymphnode and pulmonary metastases, and peritoneal seeding with bloody ascites. All mice with the implant-tumor died within 6 weeks due to serious metastasis. The 100% rate of transplantability and metastasis maintained for over 16 passages. The morphological characteristics of implant tumor cells were similar to those of the original specimen by histological and electronmicroscopic observation, and kept on secreting alpha-fetoprotein (AFP) in recipient animals. Those data from DNA content analysis by flow cytometry (D. I. value, 1.61) and chromosome karyotype revealed the existence of hypotriploid and hypertriploid cells. The results demonstrated that orthotopic implantation model of human HCC displayed features of human clinical HCC in animals. It should allow development of new treatment modalities and study of metastatic mechanism of human HCC. PMID- 9206042 TI - [Enhancement of tumor growth after partial hepatectomy and blood transfusion]. AB - Female adult BUF rats (6-8 week) received sham operation (Sham), 70% hepatectomy (PH); Sham or PH with blood transfusion (BT or PH+BT). BUF 7316A hepatoma cells were inoculated subcutaneously in the neck of rats on the operation day. Tumor size was measured from day 7 to 20 after inoculation. Sera and splenic adherent cells harvested on day 5 from Sham and PH rats were added into mixed lymphocyte cultures (MLR). The result showed that tumor growth in PH or BT rats was significantly promoted as compared to that in Sham rats (P < 0.01, P < 0.05). The most marked enhancement of tumor growth was observed in PH+BT rats (P < 0.001). Sera and splenic adherent cells from PH rat significantly inhibited MLR (P < 0.05). These results suggest that partial hepatectomy and blood transfusion are responsible for the enhancement of tumor growth. Some immunosupperassive factor might be produced in the process of liver regeneration, and blood transfusion might have an additive immunosuppressive effect. PMID- 9206043 TI - [A dynamic study of the cytotoxic effects of hyperthermia combined with cis diaminedichlorolplatinum (DDP) on human gastric cancer cell lines MKN28 and MKN45 in vitro]. AB - Cytotoxic effects of hyperthermia combined with DDP on MKN28 and MKN45 cells were studied by MTT assay according to a nested design. The rusults showed: hyperthermia alone above 43 degrees C for 30 min was cytotoxic; hyperthermia at temperature lower than 43 degrees C for 30 min could increase sensitivity of cancer cells to DDP. The cytotoxic effect of simultaneous use of hyperthermia and DDP was more marked than that of sequential use of the 2 treatments. Hyperthermia combined with DDP could inhibit growth of human gastric adenocarcinoma cells regardless of their degree of differentiation. PMID- 9206044 TI - [Lung carcinosarcoma--a report of 15 cases]. AB - Lung carcinosarcoma is a rare pulmonary mixed malignant tumor. From November 1979 to Sept. 1992, among 4251 cases of pulmonary malignant tumor operated in our department 15 were lung carcinosarcoma (0.35%). Pneumonectomy was done in 6 patients, lobectomy in 8, thoacotomy in 1. Pathologically, squamous carcinoma with fibrosarcoma was observed in 4, squamous carcinoma with chondrosarcoma in 1, squamous carcinoma with pleomorphic sarcoma in 1, adenocarcinomma with fibrosarcoma in 7, adenocarcinoma with chondrosarcoma in 1, and small-cell carcinoma with chondrosarcoma in 1. There was no operation death. Upon follow up, the 1-, 3-, 5-, and 7-year survival rates were 66.7%, 53.3%, 42.9% and 2/7, respectively. The longest survival was 97 months. The histologica origin, the relationship between pathological findings and clinical features and the diagnosis, treatment and prognosis are discussed. PMID- 9206045 TI - [Thoracoscopy in malignant pleural effusions]. AB - To assess the value of thoracoscopy in malignant pleural effusions, the procedure and results of thoracoscopy by using a fiberoptic bronchoscope and a rigid cold light thoracoscope in 130 cases with malignant pleural effusion are reported. The overall diagnostic rate was 91.5% (119/130). The malignant pleural mesothelioma in 24 cases and metastatic cancers in 95 cases were histopathologically confirmed. Talcum powder, tetracycline and Corynebacterium parvum were separately sprayed through thoracoscope into pleural cavity in 69, 10 and 10 patients, and the success rates of complete and lasting pleurodesis were 87.0%, 5/10 and 8/10 respectively. Postoperative complications included transient fever and chest pain, local subcutaneous emphysema in 6 cases and tumor seeding at thoracoscopy site in 4 cases. It is concluded that thoracoscopy is simple, safe, reliable and of high practical value in the diagnosis of malignant pleural effusions and in assessment before exploratory thoracotomy, and that transendoscopical administration of drugs for pleurodesis is a very effective method for controlling malignant pleural effusions. The efficacy of the talc poudrage is better than tetracycline and Corynebacterium parvum. PMID- 9206046 TI - [Expression and significance of hepatitis B virus genes in human primary intrahepatic cholangiocarcinoma and its surrounding tissue]. AB - In order to explore the significance of HBV in the pathogenesis of cholangiocarcinoma at molecular level, HBV DNA, X gene, pre-S gene, S gene and C gene in 40 cases of primary intrahepatic cholangiocarcinoma and its surrounding tissue were detected by in situ hybridization technique. The results showed that 33 cases (82.5%) were positive for HBV DNA, while 31 (77.5%), 26 (65.0%), 24 (60.0%) and 27 (67.5%) cases were positive for X gene, pre-S gene, S gene and C gene, respectively. The results of this study suggest that there exists a close relationship between human primary intrahepatic cholangiocarcinoma and HBV chronic persistent infection. The expression of X gene might play an important role in the pathogenesis of the primary intrahepatic cholangiocarcinoma. PMID- 9206047 TI - [A comparative study of cervical and thoracic anastomoses after esophagectomy for esophageal carcinoma]. AB - From April 1979 to December 1984, esophagectomy was performed in 552 cases of esophageal cancer of which 108 received cervical anastomosis and 444 intrathoracic anastomosis. The total postoperative complications and operative mortality rates of the two groups were very close. Leakage was significantly more frequent after cervical anastomosis, but mortality due to leakage was less frequent than that in thoracic anastomosis. The 1-, 3-, 5-, 10-year survival rates of cervical anastomosis were apparently higher than those of intrathoracic anastomosis, but the differences were not statistically significant. The 5-year survival rates of patients with the same TNM stage failed to demonstrate any significant difference between the two groups. The quality of life among the groups was satisfactory. There was no deterioration of the quality of life in cervical anastomosis. It caused less gastroesophageal reflux than did intrathoracic anastomosis. We hold that esophagectomy with cervical anastomosis and extensive lymphadenectomy is a better treatment of choice for carcinoma of the esophagus. PMID- 9206048 TI - [The most important prognostic factors for 736 patients with adenocarcinoma of the gastric cardia: a multivariate analysis using COX proportion hazard model]. AB - A prognostic study was done in 736 patients with primary resectable adenocarcinoma of the gastric cardia whose postoperative survival time was longer than three months. The 3-, 5-, and 10-year cumulative survival rate was 32.2%, 20.2% and 12.2%, respectively. Thirty individual variables were evaluated statistically using the cumulative survival rate by the computer's COX multivariate analysis model. The results showed that the most important prognostic variables for predicting overall survival included maximal tumor diameter, type of resection, depth of invasion and paracardial lymph node metastasis. Sex and preoperative radiotherapy did not influence prognosis. PMID- 9206049 TI - [The influence on prognosis of intraoperative chemotherapy for adenocarcinoma of gastric cardia]. AB - Four hundred and four patients underwent surgical treatment for adenocarcinoma of the gastric cardia in our hospital between January 1980 to June 1990. Of 342 resected cases, 231 cases were treated with surgery alone, their 1-, 3-, and 5 year survival rates were 83.8%, 38.5% and 20.8%, respectively. Since January 1987, postoperative chemotherapy was given in 47 patients, their 1-, 3-, and 5 year survival rates were 89.4%, 46.8%, 29.8%, respectively, while intraoperative chemotherapy in 54 cases, had 1-, 3-, and 5-year survival rates of 87.1%, 63.0% and 38.9%, respectively. The result showed that the 3- and 5-year survival rates were higher in postoperative chemotherapy cases than in cases with operation alone, but there was no statistical significance between the two groups. However, intraoperative chemotherapy group had much higher 3- and 5-year survival rates than operation alone (P < 0.01). Intra-operative chemotherapy is recommended especially in patients beyond stage I with lymph node metastasis. PMID- 9206050 TI - [Computed tomographic features of malignant fibrous histiocytoma]. AB - CT findings in 45 cases of malignant fibrous histiocytoma (MFH) were reviewed. There were 27 primary and 22 (23 episodes) cases of recurrent tumor, 4 (5 episodes) cases of recurrent tumor were initially treated in our hospital. Tumors located in craniofacial area in 16 cases, abdomen or retroperitoneum in 17, soft tissues of extremities or trunk in 12. There were 26 male and 19 female patients, with 19-82 years of age (medium, 46 years of age). The most common clinical symptom was a local mass (64.4%). On CT scan, 84% (42/50) of lesions with tumor mass > 5 cm, mostly with irregular contour, medium to markedly hyper-attenuated enhancement were revealed in 78.8% of cases and 60.6% of cases with hypo attenuated necrotic area inside. Most of the tumors invaded adjacent organs or structures which were revealed in 75.8% of cases, such as infratemporal fossa, pterygopalatine fossa, extradural space in the craniofacial lesions; liver, psoas muscle, abdominal wall in abdominal and retroperitoneal lesions. Expansible change could be shown when paranasal sinus was invaded (8/9). The CT findings of recurrent lesions were similar to the primary ones. Enhanced CT scan may provide useful information in demonstrating the nature and extent of invasion of the tumor. Closely integrating CT and pathologic findings is important for the differential diagnosis of malignant tumors of soft tissue. PMID- 9206051 TI - [The effect of long-term tamoxifen therapy on the occurrence of contralateral primary breast cancer]. AB - From January 1 1985 to December 31 1990, 874 cases of female primary breast cancer were treated in the Department of Surgery at Beijing Institute for Cancer Research. Of these, 21 patients suffered from contralateral primary breast cancer after surgery. These patients were divided into two groups, 523 patients received adjuvant tamoxifen therapy (20mg daily) for 2 to 5 years as the treated group. There were 351 patients without tamoxifen therapy as the control group. The medium follow-up of the treated and the control group was 7.8 years and 7.0 years, respectively. The incidence of contralateral primary breast cancer in the treated group was 1.5% (8/523), and that in the control group was 3.7% (13/351, P < 0.05). This result suggests that tamoxifen is useful to reduce the risk of contralateral primary breast cancer. PMID- 9206052 TI - [A clinico-pathologic study of primary malignant fibrous histiocytoma of bone]. AB - Primary malignant fibrous histiocytoma of bone (PBMFH) is a rare tumor. In this series 11 cases of PBMFH (7 men, 4 women, mean age 39.6 years, range 15-62) were reported. They were studied by using morphology, immuno-histochemistry combined with the relevant clinical materials. It showed that most of the patients complained of local pain as the first symptom, and it generally arose in proximal or distal end of extremities. Roentgenographically, it mainly displayed a lytic destruction of bone. Like soft tissue malignant fibrous histiocytoma, it was composed of a mixture of plump fibroblast-like cells and large round or polymorphic histiocyte-like cells and usually showed a storiform pattern. But there were no myxoid and inflammatory subtypes encountered in these cases, except the storiform-pleomorphic type (9 cases) and giant cell type (2 cases). So it appears to have some difference between the subtypes of MFH of bone and soft tissue. Since the histomorphology and immunohistochemical reaction are less characteristic, especially with the morphological pleomorphism and marked cellular heterogeneity, diagnosis of PBMFH must depend on multifacet observation and comprehensive assessment. PMID- 9206053 TI - [Classification of ninety-eight adult cases of acute leukemia according to morphology, immunology and cytogenetics]. AB - Ninety-eight cases of adult acute leukemia (AL) were diagnosed and classified based on morphologic, immunologic and cytogenetic (MIC) features. The results showed that: the conformity rate of cytomorphologic/cytochemical classification with MIC classification was 90.8%. For ALL, the conformity rate of immunologic classification with MIC classification was 95.6%, but it was only 70.8% for AML. Of the 48 AML, 10 expressed lymphoid lineage, associated antigens and 8 of 43 ALL expressed myeloid lineage-associated antigens. Seven cases were diagnosed as hybrid acute leukemia according to Catovsky criterion. The chromosome aberrations were found in 70 cases, of them 46 cases showed characteristic abnormalities including t(9;22), t(4;11), t(11;14), t(8;12), t(8;14), 6q-, 9p-, and t(15;17), t(8;21), inv(16), etc. PMID- 9206054 TI - [A randomized trial of tropisetron in the prophylaxis of nausea and vomiting induced by chemotherapy]. AB - Thirty patients receiving cisplation or non-cisplatin (containing cyclophosphamide and adriamycin) chemotherapy were enrolled in a randomized, crossover study comparing the efficacy of single dose of Navoban (tropisetron, 5 mg) and Kytril (granisetron, 3 mg). The effective control of acute vomiting induced by cisplatin was achieved in 95.2% (20/21) of patients receiving Navoban and 90.5% (19/21) in those receiving Kytril. Complele control rate was 71.4% (15/21) in Navoban arm, and 81.0% (17/21) in Kytril arm. Total control of delayed vomiting (day 2-5) was 71.4%-90.4% in Navoban arm, while it was 66.7%-4% in Kytril arm. The effective control of vomiting induced by non-cisplatin drugs was achieved in 9/9 in both arms. It is concluded that both agents are effective in the control of vomiting induced by chemotherapy. They have identical adverse effects and are well tolerated by the patients. PMID- 9206055 TI - Chemotherapy of parasitic diseases. Current status and new directions. PMID- 9206056 TI - Emergency percutaneous transluminal coronary angioplasty in acute myocardial infarction complicated with cardiogenic shock. AB - OBJECTIVE: To approach the efficacy of emergency percutaneous transluminal coronary angioplasty (PTCA) in Chinese patients with acute myocardial infarction (MI) complicated by cardiogenic shock. METHODS: Twelve patients with cardiogenic shock treated by emergency PTCA in this institution from January 1990 to May 1994 were retrospectively reviewed. There were anterio-lateral MI in 4 cases, inferio posterial MI in 7, anterial and inferial MI in I, of which triple vessel disease in 4 cases, double vessel disease in 6, single vessel disease in I and left main coronary artery (LM) disease in 1. PTCA was performed under assistance of IABP in 7 cases and of centrifugal pump in 1. Only infarct-related Coronary artery (IRCA) was dilated during acute phase. Direct PTCA was done in 7 cases and rescue PTCA after failure of thrombolytic therapy in 5. RESULTS: PTCA was successful in 11 (91.7%) of the 12 cases, in 1 case with failure of PTCA the IRCA was reperfused by intracoronary thrombolytic therapy. Cardiogenic shock was reverted in 7 (63.6%) of the 11 cases reperfused by PTCA, among whom 6 (54.5%) survived. Four died of irreversible shock after 3 hours-9 days and 1 case with reversed shock died of cardiac rupture after 4 days. During a period of 13-55 months follow-up, long-term survivals were obtained in 4 cases. CONCLUSIONS: Emergency PTCA can achieve a high success rate and significantly decrease the mortality rate to less than 50% in Chinese patients with acute MI complicated with cardiogenic shock. The patients tolerate the PTCA procedure very well under the assistance of intra aortic balloon counterpulsation (IABP) even in very severe condition. PMID- 9206057 TI - Beneficial effects of captopril on prognosis in patients with acute myocardial infarction. Shanghai Secondary Prevention of Acute Myocardial Infarction Study Group. AB - OBJECTIVES: To assess the effects of early and long-term angiotensin-converting enzyme inhibitor treatment with captopril on clinical outcome in patients with acute myocardial infarction (AMI). METHODS: Eight hundred and twenty-two patients with AMI who were hospitalised within 72 hours of symptoms and had no cardiogenic shock were randomly allocated to captopril (n = 478, Group I) and conventional treatment (n = 344, Group II). Cardiac events including congestive heart failure, reinfarction, severe arrhythmias and cardiac death during hospitalization and follow-up period (average 20 months) were determined. RESULTS: The overall mortality rate during hospitalization was lower in group I than in group II (P = 0.0001), this was true for patients with anterior (P = 0.0003), inferior (P = 0.0411) and anterior inferior AMI (P = 0.0232). During follow-up, despite similar occurrence rate of reinfarction and severe arrhythmias in the two groups, the mortality rate (P = 0.0324) and total cardiac event rate (P = 0.055) were lower in group I than in group II. CONCLUSIONS: After AMI, early and long-term treatment with captopril exerts a beneficial effect on the prognosis of patients. PMID- 9206058 TI - Long-term oral administration of L-arginine enhances endothelium-dependent vasorelaxation and inhibits neointimal thickening after endothelial denudation in rats. AB - OBJECTIVE: To further prove the hypothesis that local decrease of nitric oxide (NO) synthesis and/or its activity might be critically important in the disturbance of vascular homeostasis after vascular injuries. METHODS: Intimal thickening model induced by air-drying denudation of rat right common carotid artery was performed to evaluate the effects of long-term oral administration of L-arginine on neointimal thickening and acetylcholine-induced endothelium dependent vasorelaxation (EDR) by histomorphometric and functional studies. RESULTS: Reductions in EDR function persisted and simultaneously developed prominent neointimal thickening by 14 days after denudation. Long-term oral supplementation of L-arginine (1 g/kg/day) significantly enhanced EDR from 43.5% +/- 12.35% to 68.8% +/- 9.0% (n = 10, P < 0.001) and reduced neointimal thickening from 62.45 microns +/- 11.26 microns to 21.45 microns +/- 6.34 microns (n = 10, P < 0.001) as compared with each control animals. CONCLUSIONS: This study shows that oral administration of L-arginine significantly inhibits neointimal thickening and preserves NO-mediated EDR in experimental endothelial denudation, suggesting an important role for L-arginine, NO pathway in the regulation of vascular homeostasis after endothelial injury which might be salutary in prevention restenosis after coronary angioplasty. PMID- 9206059 TI - Prevalence and incidence of NIDDM in Daqing City. AB - OBJECTIVE: To investigate the prevalence and incidence of non-insulin dependent diabetes (NIDDM) in China by WHO criteria. METHODS: In the prevalence survey of NIDDM all 110660 participants (55391 men, 53269 women) were inhabitants of Daqing City, the largest oil center in northeast China, accounting for 87.3% of the 25 to 74 aged population in this city. They were screened by measuring two-hour plasma glucose concentrations (PG2 h) after a breakfast containing at least 80 g of carbohydrate, then a standard oral glucose tolerance test (OGTT, 75 g glucose load) was performed in 4209 subjects with PG2 h more than 6.67 mmol/L in this screen. NIDDM and impaired glucose tolerance (IGT) were diagnosed using WHO criteria. Incidence survey was made in 36471 non-diabetics identified in the prevalence survey. Two-hour urine-glucose after breakfast was determined during first screen. The urine-glucose trace or positive subjects were then followed by OGTT. Glucose was measured by glucose oxidation method. RESULTS AND CONCLUSIONS: In prevalence survey, 630 newly diagnosed NIDDM (296 males, 334 females) and 596 IGT (318 males, 278 females) were found in 110660 (male:female = 55391/53269) studied subjects in addition to 190 previously diagnosed NIDDM. Thus the total prevalence of NIDDM was 7.7/1000, and IGT was 5.5/1000. Standardized to the Chinese population in 1982, the prevalences are 12.6/1000 (95% CI = 12.0/1000 13.3/1000) and 7.7/1000 (95% CI = 7.16/1000-8.19/1000) respectively. In the incidence survey, 191 NIDDM (103 males, 88 females) were diagnosed in the 36471 (male:female = 18801/17670) non-diabetics from 1986 to 1990, thus the annual incidence of NIDDM was 131/100000 (137 males, 125 females). Standardized incidence is 131/100000 (95% CI = 94/100000-168/ 100000). It is estimated that there would be more than 700 thousand new diabetics per year in 24-74 years old Chinese if Chinese population were 1.3 billion in the early 21st century. PMID- 9206060 TI - The role of GRH mediated AC-cAMP system in the pathogenesis of human pituitary GH secreting adenomas. AB - OBJECTIVE: To investigate the role of AC-cAMP system in the transmission of the action of the growth hormone releasing hormone (GRH) on growth hormone (GH) release in pituitary GH-secreting adenomas. METHODS: The effects of GRH (10(-7) mol/L) on intracellular cAMP levels and GH release and the effects of AC-cAMP stimulators, cholera toxin (Ct, 50 micrograms/L), forskolin (10(-5) ml/L) and db cAMP (10(-3) mol/L) on GH secretion were studied in cultured cells of 21 GH secreting adenomas obtained from operation for acromegalic patients. RESULTS: GRH and Ct failed to stimulate GH secretion in 61.9% (13/21 cases) and 57.1% (12/21 cases) pituitary GH adenoma cell cultures respectively. Forskolin stimulated GH release in 88.9% (8/9 cases), while db-cAMP induced GH secretion in all cases tested (5/5 cases). The intracellular cAMP levels were elevated by GRH in the 4 out of 9 cases of tumor cell cultures, but not in the other 5 cases. According to the GH secretory responses to GRH and Ct, the 21 GH tumors were divided into 4 groups. In group A and B, GRH can stimulate GH release, but Ct has stimulative role only in group A. In group C and D, GRH fails to stimulate GH secretion. However group A can respond to Ct, but group D has no response. CONCLUSIONS: The GH hypersecretion in most acromegalic patients is mainly due to the defects of pituitary adenoma cells, especially the abnormalities of GRH receptor and/or stimulative guanosine protein. PMID- 9206061 TI - Influence of lipopolysaccharide and interleukin-2 on proliferation and synthesis of sulfated macromolecules in cultured rat glomerular epithelial cells. AB - OBJECTIVE: To investigate the effects of lipopolysaccharide (LPS) and human recombinant interleukin-2 (hrIL-2) on the synthesis of sulfated macromolecules in cultured rat GECs. METHODS: With 3H-thymidine (3H-TdR) and 35S-Na2SO4 dual isotope labelling technique, the proliferation and synthesis of sulfated macromolecules in cultured rat GECs were estimated according to the 3H-TdR uptake and 35SO4(2-)-contents in cell layers respectively. RESULTS: Both LPS (6.25-50.0 micrograms/L) and hrIL-2 (2.5-10.0 kU/L) decreased the uptake of 3H-TdR by GECs (P < 0.05). As for incorporation of 35SO4(2-) into cell layers, the results that both means and adjusted means in hrIL-2 groups were lower than those in controls (P < 0.05) implied that hrIL-2 could inhibit the synthesis of basement membrane HSPG while LPS hindered incorporation through its inhibitory effect on cell proliferation. CONCLUSIONS: We may infer from these results that LPS-like proinflammatory mediators could increase the subpopulation of large pores in capillary wall through the reduction of cell-associated HSPG, and IL-2-like cytokines could contribute to the development of proteinuria through its inhibitory effect on the production of basement membrane HSPG in immune-mediated glomerular disorders. PMID- 9206062 TI - A study of platelet specific antibody (anti-Sib(a)). AB - OBJECTIVE: To screen and analyse the platelet-specific alloantibody form the sera of patients who received multiple platelet transfusions or experienced side effects after platelet transfusions. METHODS: 156 patients, who received multiple platelet transfusion or experienced side effects after platelet transfusion, were collected from hospitals in Shanghai. The simplified sensitized erythrocyte platelet serology assay (SEPSA) was used to screen the platelet antibodies and the identification of specificities of the platelet alloantibodies was performed with a platelet panel of known antigens and absorption tests. RESULTS: Seventy four sera were found positive. Among them 59 sera (47.4%) contained HLA antibodies, two sera (2.7%) platelet-specific antibodies, and 13 sera both HLA and platelet-specific antibodies. One of 2 sera containing platelet-specific alloantibody, the serum Zhu, was confirmed to have a monospecific platelet specific alloantibody, the anti-Sib(a) (HPA-2b). CONCLUSIONS: We report the first case of anti-Sib(a) in Chinese Han population. The antigen frequency of Sib(a) is considered as one of the most important antibodies which will cause the platelet transfusion refractoriness in China. PMID- 9206063 TI - Percutaneous transluminal balloon pulmonary valvuloplasty using domestic balloon catheter for congenital pulmonary valve stenosis in children. AB - OBJECTIVE: To estimate the effect of percutaneous balloon pulmonary valvuloplasty (PBPV) using domestic balloon catheter for congenital pulmonary valve stenosis (PVS) in infants and children. PATIENTS AND METHODS: The data of 70 patients aged from 1.5 to 12 years (mean 5.5 +/- 3.4 years) who underwent PBPV using domestic balloon catheter for PVS in our institution were reviewed. Sixty-six patients had dome-shaped valve stenosis, and 4 had displastic valve stenosis. SaO2 reduced in 5 patients and associated open foramen ovale was noted in 23. Predilatation, right ventricular systolic pressure ranged from 60 to 234 mmHg (mean 115.75 +/- 36.15 mmHg). The systolic gradient (delta P) from right ventricle to pulmonary artery was 89.92 +/- 38.25 mmHg. Balloon diameter was selected 120%-142% of pulmonary valve annulus diameter (mean 132%). All patients were followed up for 3 months-4 years by means of clinical examination. ECG, 2D-UCG, and Doppler-UCG. RESULTS: After dilatation, delta P reduced from 89.92 +/- 38.25 to 14.65 +/- 11.40 mmHg (P < 0.001). The rate of decrease in transvalve gradient (delta P) was 83.7%. No patients experienced procedure-related events. Mid-term follow-up showed that no patients had clinical and instrumental complications. Doppler and ECG analyses showed that delta P did not change significantly similar to immediate gradient after PBPV and right ventricular hypertrophy disappeared progressively 6 months after PBPV. CONCLUSIONS: PBPV is a useful, safe and definitive procedure in the treatment of isolated PVS in infants and children. The properties and features of the balloon catheter produced by Shanghai Med-Tech Factory are similar to those of Med-Tech balloon catheter of U.S.A. PMID- 9206064 TI - Endoscopic ultrasonography assessment in preoperative staging for carcinoma of ampulla of Vater and extrahepatic bile duct. AB - OBJECTIVE: To evaluate preoperatively the extent of primary tumor, involvement of regional lymph node, and distant metastasis of ampullary carcinoma and extra hepatic bile duct carcinoma. METHODS: 28 patients with ampullary carcinoma and the 18 patients with extrahepatic bile duct carcinoma were subjected to endoscopic ultrasonography (EUS). The results were compared with those of surgical explorations and pathological findings of the resected specimens for evaluating the accuracy of preoperative staging of EUS. 46 patients underwent surgical explorations. Radical resection with detailed pathological study was done for 22 patients with resectable ampullary carcinoma and 18 patients with extrahepatic bile duct carcinoma. Carcinomas of ampulla of Vater and extrahepatic bile duct were staged according to the tumor, nodes, metastasis (TNM) classification. RESULTS: The accurate rate of EUS in assessing the extent of cancer invasion was 81.8% for ampullary carcinoma, and 72.2% for extrahepatic bile duct carcinoma. The accuracy of EUS in predicting regional lymph node metastasis was 59% for ampullary carcinoma, and 61.1% for extrahepatic bile duct carcinoma. Invasion of portal vein was correctly predicted by EUS in 2 of 3 patients, but the 3 cases of liver metastasis were not detected by EUS. CONCLUSIONS: EUS is an effective method for the evaluation of the extent of invasion of ampullary carcinoma and extrahepatic bile duct carcinoma as well as the involvement of regional lymph node before operation. Because of its limited penetration depth, however, EUS is inadequate in the assessment of liver metastasis. PMID- 9206065 TI - Endovascular embolization of intracranial aneurysms with self-made tungsten coils in a dog model. AB - OBJECTIVE: To evaluate the embolization effects and the long-term histologic changes, including the ultrastructure of the neoendothelium, in experimental canine aneurysms obliterated with self-made tungsten microcoil. METHODS: Twenty eight experimental aneurysms were microsurgically created in 14 mongrel dogs by using end-to-side jungular carotid anastomosis. The aneurysms were obliterated with either self-made tungsten microcoil alone or the microcoil plus micro direct current electrocoagulation, with preservation of the parent vessel. The animals were kept in observation. 48 hours, 4 weeks, and 4 months after the embolization of the aneurysms, repeated carotid arteriography was performed to assess the potential recanalization of the aneurysms. The animals were then respectively killed and submitted for autopsy. The carotid artery and the embolized aneurysms were resected and studied with light and electron microscopy. RESULTS: A total of 14 obliterated aneurysms were completely excluded from the parent circulation by an endothelialized layer of connective tissue. The fundus of the aneurysm was completely obliterated by heavy reactive fibrous tissue surrounding the microcoils with very minimal inflammatory reaction. CONCLUSIONS: Tungsten microcoils are a relatively ideal embolic material at present for endovascular treatment of aneurysms. Microcoils plus micro direct current electrocoagulation yield more reliable, accurate, and safer embolic effects than microcoils alone for the obliteration of aneurysms. PMID- 9206066 TI - Cryopreservation of embryos in an IVF-ET program. AB - OBJECTIVE: To evaluate the use of cryopreservation of embryos in an in vitro fertilization and embryo transfer (IVF-ET) program. METHODS: Three cases of successful pregnancy after frozen-thawed embryo transfer were reviewed. The Testart's slow freezing and rapid thawing method was chosen. RESULTS: The first case was a Turner's syndrome patient and her husband had azoospermia. Donated frozen-thawed embryo transfer was done after hormonal replacement therapy. Pregnancy was achieved and a normal baby was delivered. The other two cases of IVF-ET did not achieve pregnancy during the stimulated cycle but became pregnant with frozen-thawed embryos in the natural cycle 4-5 months later. CONCLUSIONS: Pregnancy could be achieved in couples with incurable reproductive defects with donated embryos cryopreserved and ready for use whenever the endometrium is prepared by hormonal replacement therapy to be synchronous with the embryo development. In an IVF-ET program, one egg-retrieval may serve for many future chances of embryo transfer and therefore increase the cumulative pregnancy success rate. PMID- 9206067 TI - A cytogenetic study of 514 Chinese couples with recurrent spontaneous abortion. AB - OBJECTIVE: To study the role of chromosomal aberration in the causation of recurrent spontaneous abortion (RSA) in Chinese population. METHODS: A total of 514 Chinese couples with 2 or more spontaneous abortions at less than 24 weeks of gestation were included. For each proband, a minimum of 13 metaphases were analyzed by G-banding. Additional cells (usually 50-100 cells) were screened when mosaicism was suspected. Chi 2 test was used to compare the number and frequency of couples with and without balanced translocation with respect to whether liveborn was present or absent. Chi 2 test for trend was used to show whether a correlation existed between the occurrence of balanced translocation and the number of spontaneous abortions at ascertainment. RESULTS: The overall incidence of chromosome anomaly was 51 out of 514 (9.92%). Chi 2 test for trend analysis showed that the chance of one member of a couple being a balanced carrier increased with the number of spontaneous abortions. The chance of finding translocation in couples with liveborn was higher than that in couples without liveborn, but the difference was not statistically significant. We also found that pericentric inversion 9 did not play an important role in the causation of recurrent abortion. CONCLUSIONS: Cytogenetic analysis is indicated in couples with 2 or more spontaneous abortions and the chance of finding chromosomal aberration increases with the number of abortions at the time of ascertainment. PMID- 9206068 TI - An observation on otoacoustic emission and ultrastructure of cochlea in experimental autoimmune inner ear disease. AB - OBJECTIVE: To acquire the detailed knowledge of immune lesion of inner ear. METHODS: A simultaneous study on physiology and ultrastructure was carried out in guinea pigs immunized with homogenous inner ear antigen. The specific antibody was analysed by a modified immunotransfer electrophoresis method. RESULTS: Three positive bands (with molecular weights of 68.0 kD, 62.7 kD and 60.1 kD) were found in all the sera of immunized animals but not in the control animals. Threshold shift of auditory brainstem response over 10dB HL was found in 42% of secondary system immunized animals and 32% of secondary endolymphatic sac (e.sac) immunized animals. Amplitude of distortion production otoacoustic emission dropped obviously in the above two groups. Transmission electron microscope showed the mitochondria of outer hair cells in the cochlea was impaired in animals with auditory disorder. CONCLUSIONS: The research suggests that the immune reaction against homogenous inner ear antigen results in dysfunction of hydroxycorticosteroid during the early stage. PMID- 9206070 TI - Radiofrequency ablation to cure atrial fibrillation: mechanisms and strategies. PMID- 9206069 TI - A clinical study on severe hydroa vacciniforme. AB - OBJECTIVE: To describe a severe type of hydroa vacciniforme (HV) in order to provide basis for treatment and prevention of this disease. METHODS: Nine cases of HV were collected in our outpatient department, and clinical and laboratory examinations were performed. RESULTS: All the cases had erythemas, papules, bullae, erosions or ulcers on exposed sites, with recurrences, gradually resulting in deformation. Five patients suffered from hand deformation manifesting rigidity, flexor tetanus, or malposition of the first, second and third interphalangeal joints of hands. Two of the above mentioned patients manifested partial bone absorption of fingers. Five patients showed partial defect of auricle. Two patients showed saddle nose, and partial absorption of nasal cartilage. One patient displayed cicatricial contracture of lower lip, with incisor extrusion. Four patients showed opacity of cornea. Through laboratory examinations, porphyrinopathies were excluded. Phototests showed increased sensitivity to ultraviolet-A radiation (UVA). CONCLUSIONS: Based on the clinical features and laboratory examinations, these patients were diagnosed as having severe HV. It is believed that this condition belongs to a disease spectrum induced by ultraviolet light. Our analysis suggests that for children with severe type of HV, preventive measures and therapeutic agents should be taken as soon as possible in order to avoid deterioration of the disease and malformations. PMID- 9206071 TI - Diagnostic key point by enteroclysis in a case of inverted Meckel's diverticulum. PMID- 9206072 TI - Orthopaedics in China: its past and present. PMID- 9206074 TI - Limb salvage in primary malignant bone tumors by intraoperative microwave heat treatment. AB - OBJECTIVE: To use intraoperative microwave heat for the treatment of tumor bearing bone in situ for limb-salvage in primary malignant bone tumor. PATIENTS AND METHODS: Twenty-five patients with malignant bone tumor received surgical protocol for limb-salvage by wide exposure and intraoperative microwave heat treatment of tumor bone in situ. The tumor was localized at distal femur in 16 patients, proximal tibia in 5, shaft of femur in 2, and illia in 2. RESULTS: The pathological diagnosis revealed osteosarcoma in 16 patients, parosteal osteosarcoma in 4, chondrosarcoma in 3, and leiomyosarcoma in 2. The stage of tumor was classified into IIB in 16 patients, IIA in 6, and IIIB in 3. The operative technique was wide exposure for tumor in soft tissues, protecting the surrounding normal tissue from heat injury by copper mesh, and heating tumor bearing bone at 50 degrees C for 15 minutes. The patients were followed up from 4 to 180 months (mean, 63 months). The survival rate of 2, 5 and 10 years was 81.3%, 70.4%, and 53.8% respectively. The functional results were less than 50% in 4 patients, between 50% and 75% in 9, and more than 75% in 12, referred to the normal function of a normal limb. The complications consisted of infection in 4 patients, pathological fracture in 4, and separation of epiphysis in 1. Oncological results showed that local recurrences of tumor were in 4 patients, and 6 patients suffered from distant metastasis. CONCLUSION: The treatment is an alternative to replacement by prosthesis or allograft bridging techniques. PMID- 9206073 TI - Surgical treatment of bone tumors in conjunction with microwave-induced hyperthermia and adjuvant immunotherapy. A preliminary report. AB - OBJECTIVE: To develop an alternative approach in conjunction with microwave induced hyperthermia. PATIENTS AND METHODS: Thermotherapy with microwave intracorporeal irradiation was used to treat 73 patients with bone tumors. The series was composed of 58 patients with malignant tumors and 15 with benign tumors: most of tumors occurred about knee joints (53/73 = 72.6%). The surgical procedure included separating the tumor bearing segment from surrounding normal tissues with a safe margin, cooling the normal tissues including the neurovascular bundle and the intraarticular structures with a water circulation system, while heating the tumor with the antenna array of a microwave system and providing an adequate soft-tissue cover for the dead bone. Postoperatively, an immune therapy regimen was carried out regularly. The patients' immunologic functions were monitored by assay of the subpopulation of T cells, IL-2 and sIL-2 R (soluble IL-2 receptor). RESULTS: Follow-up varied from 3 to 38 months (mean 19 months). Excluding 3 patients with malignancy in the vertebrae treated for palliation, 70 were evaluated according to oncological and orthopedic criteria. Five patients had local recurrence and required amputation. The remaining 65 had excellent local control. In 6 of the 55 patients with malignancy of the extremities, lung metastasis occurred one to two years after surgery. The oncological results were similar to those obtained by other limb-saving procedures. Pathological fracture occurred at devitalized bone in 5 patients. In 72.5% of the patients (29 of 40 tumor-free cases followed more than one year), knee joints functioned well, being stable and painless with almost full range of motion. Single photon emission computered tomography (SPECT) for 16 patients revealed revascularization of the devitalized tumor bearing bone segment could accomplish in one year or more. The immune states were improved in various extends after thermotherapy plus immunotherapy in the majority of patients. CONCLUSION: These results show that the use of microwave hyperthermia and adjuvant immunotherapy in conjunction with the surgical treatment of bone tumors can be considered a definitive procedure, which is safe and well-tolerated. The oncological and orthopedic results are encouraging. PMID- 9206076 TI - Vascularized iliac periosteal transfer for the treatment of avascular necrosis of the femoral head and a new evaluation grading system. AB - OBJECTIVE: To introduce the surgical technique and a new evaluation grading system and to report the results of the vascular iliac periosteal grafting for avascular necrosis of the femoral head. PATIENTS AND METHODS: From 1983 to 1994, a new technique was used to treat 75 hips of 60 patients with avascular necrosis (AVN) of the femoral head using the vascular iliac periosteum with deep iliac circumflex artery and vein pedicle. 52 patients (66 hips) were followed up for 3 11 years and all patients were evaluated clinically and roentgenographically according to the new evaluation grading system for AVN by the Chinese Society for Osteonecrosis. RESULTS: Preoperatively, the average AVN score was 57 pts., and all patients had significant pain and limp. Five hips were at Ficat/Arlet stage IV, 40 were at stage III, and 21 were at stage II. Postoperatively, the average AVN score was 83 pts., 47 hips were pain free, 13 had mild pain, and 6 had moderate pain. At the latest follow-up, according to the new evaluation grading system, 26 hips were graded as excellent, 32 hips as good, 5 hips as fair, and 3 hips as poor. CONCLUSION: It is one of the better alternatives that can prevent the necrotic femoral head from progressing to collapse and promote revascularization and new bone formation by a direct mechanism, and it is suitable for the treatment of AVN of the stage I to III. PMID- 9206075 TI - Transposition of flexor pollicis brevis muscle for reconstruction of thumb opposition. Anatomical study and clinical application. AB - OBJECTIVE: Based on the anatomical study and clinical trial of transposition of the flexor pollicis brevis muscle (M. f. p.b.) for the reconstruction of thumb opposition, we suggested a new method for the treatment of irreparable median nerve injury which causes paralysis of the opponens pollicis and the abductor pollicis brevis muscles (M.a.p.b) and leads to loss of thumb opposition. MATERIAL AND METHODS: Anatomical study and biomechanic analysis were performed on 20 cadaveric hands and 8 patients who had been treated and followed up on an average for 12 months. RESULTS: The M.f.p.b. overlaps the M.a.p.b. for about half of its width at the muscle origins and the overlapping reduced to about 1/3 of the width at the muscle belley level. The M.f.p.b. chiefly inserted on the palmar aspect of the base of proximal phalanx. The M.a.p.b. primarily was inserted on the radial side of the first metacarpophalangeal(MP) joint, and the M.f.p.b. was entirely innervated by the deep branch of the ulnar nerve. In an attempt to increase the angle between longitudinal force lines of these two muscles for 7 degrees-9 degrees, we transferred the insertion of the M.f.p.b. to the radial side of the MP joint, so that it gives this muscle the function of opposition. Eight patients were treated and followed up on an average for 12 months. All had fine functional results. CONCLUSIONS: This method is effective, and least traumatic, and does not need transposition of another tendon. PMID- 9206077 TI - Presence of free radicals in pigment gallstone in vivo. AB - OBJECTIVE: To clarify whether free radical, which may play a role in pigment gallstone formation, is present in pigment gallstones in vivo. MATERIALS AND METHODS: Free radical signal of gallstones from 18 patients was detected by electron paramagnetic resonance spectroscopy at 77K under anaerobic condition and in air (control). As soon as the anaerobic determination was finished, the fresh anaerobic sample was exposed to air and stored in a freezer at -20 degrees C. RESULTS: Free radical signal (g = 2.0038) was detected in fresh anaerobic samples containing more than 2% bilirubin compound, and the signal intensity correlated linearly with the content of calcium bilirubinate (r = +0.95, P < 0.0005). During the storage at -20 degrees C and exposure to air, the signal intensity of each anaerobic sample and its control increased gradually, eventually reaching the same stable level. Fe(III) signal intensity was enhanced synchronously and related linearly with free radical signal (r = +0.99, P < 0.0005). CONCLUSIONS: Free radical exists originally in pigment gallstones in vivo, and it may play an important role in pigment gallstone formation. The free radical signal carried by gallstones may be strengthened by the action of oxygen in air on bilirubin. The transition metal ions probably take part in the formation of bilirubin free radical. PMID- 9206078 TI - Incremental iron overload during reperfusion progressively augments oxidative injury. AB - OBJECTIVE: To determine if a relationship exists between the extent of iron catalyzed injury and the degree of tissue iron overload during reperfusion. METHODS: To selectively increase tissue iron only during early reperfusion, isolated, buffer perfused rabbit hearts were exposed to 20 microM Fe(2+)-100 microM ADP during the last 3 minutes of ischemia and the initial 4 minutes of reperfusion. Control groups were exposed to ADP and iron-ADP regimens that did not increase intracellular iron. All the hearts received 30 minutes of normothermic global ischemia and 30 minutes of reperfusion. Heart function was monitored continuously throughout each experiment. Tissue iron and biochemical markers were analyzed at the end of experiments. RESULTS: Hemodynamic recovery was decreased and tissue lipid peroxide levels were increased in the 20 microM Fe(2+)-100 microM ADP group compared to controls. The recoveries of developed pressure and positive/negative dP/dT at 30 minutes of reperfusion were negatively correlated with tissue iron levels, while cytosol and membrane lipid peroxide levels correlated positively with the iron levels during reperfusion. CONCLUSION: The extent of oxidative injury during reperfusion was directly related to the tissue iron burden present during reperfusion. Increased lipid peroxidation was the principal chemical marker of iron-catalyzed injury. PMID- 9206079 TI - Exercise hemodynamic benefits of rate adaptive ventricular pacing. AB - OBJECTIVE: To investigate the exercise hemodynamic benefits of activity-sensing rate adaptive ventricular pacing (VVIR) over fixed rate pacing (VVI) mode. METHODS: Activity sensing rate adaptive pacemaker was implanted in 19 patients (13 males and 6 females, mean age 54.8 years) with bradycardia. All patients underwent symptom-limited upright bicycle exercise in VVIR and VVI pacing modes in random order after implantation. With electrocardiogram monitor and M-mode echocardiography, heart rate, stroke volume and cardiac output were measured at rest and at each stage of exercise. RESULTS: All patients were pacemaker dependent, without any spontaneous heart rhythm throughout this study. In the activity sensing ventricular pacing mode, all patients achieved a significant increase in exercise duration compared to fixed rate ventricular pacing mode (mean +/- s, 437 +/- 45 vs 323 +/- 23sec; P < 0.01), with a mean maximum pacing rate of 113 +/- 23ppm. Although the cardiac output was significantly improved in both pacing modes (10.2 +/- 1.4L/min with VVIR and 7.5 +/- 1.1L/min with VVI), the maximum exercise cardiac output in VVIR was increased over VVI by 46% (P < 0.05). Additionally, the stroke volume was significantly increased by 50% or more at rest in VVI mode, but was relatively maintained in VVIR mode (P > 0.05). CONCLUSION: Rate adaptive ventricular pacing can significantly improve the exercise capacity and cardiac output in patients with bradycardia. The increment of exercise cardiac output in VVIR mode is mainly dependent upon the pacing rate during exercise. PMID- 9206080 TI - Effects of selective head cooling on brain cell membrane activity during postischemic reperfusion. AB - OBJECTIVE: To examine the effect of selective head cooling (SHC) on brain cell membrane activity involving ATPase, phospholipase A2, content of total membrane phospholipids during postischemic reperfusion, so as to elucidate the possible underlying mechanism on resuscitating effect of SHC. MATERIALS AND METHODS: Complete cerebral ischemia (CCI) was induced by the four-vessel model. 56 New Zealand rabbits were allocated randomly into two groups, non-ischemic control group had 30, 180 and 360 minutes reperfusion after CCI (n = 8); and SHC group with the same ischemic-reperfusion insult were all treated with SHC (28 degrees C, surface cooling method). Changes of Na+, K(+)-ATPase, Ca2+, Mg(2+)-ATPase, phospholipase A2, total phospholipids of brain cell membrane were observed. Comparison of data between two groups was made by Students' t test. RESULTS: Compared with non-ischemic controls following 30 minutes CCI, activities of Na+, K(+)-ATPase stepwisely decreased at 30, 180 and 360 minutes, Ca2+, Mg(2+)-ATPase dropped at 180 and 360 minutes, phospholipidase A2 increased markedly at 30, 180, 360 minutes, and total phospholipids decreased at 180 and 360 minutes reperfusion (P < 0.01). Selective head cooling inhibited all the above changes significantly (P < 0.01). CONCLUSION: The results suggest that selective head cooling initiated soon after reperfusion is beneficial for brain cell membrane function recruitment, which provides favourable effects on the damaged but still remediable brain cells for their resuscitation. PMID- 9206081 TI - Effects of MTX and BN52021 on PAF-induced chemotaxis of PMNs and intraepidermal accumulation of inflammatory cells in guinea pigs. AB - OBJECTIVE: To investigate the effects of methotrexate (MTX) and platelet activating factor (PAF) antagonist ginkgolide B (BN52021) on PAF induced chemotaxis of neutrophils. MATERIALS AND METHODS: All guinea pigs were randomly divided into 12 groups. They were given different dosages of MTX and BN52021 by intra-abdominal injections. The random and chemotactic migration of polymorphonuclear leukocytes (PMNs) were measured by the agarose method. The backs of all guinea pigs were given intradermal injections of PAF and the numbers of the infiltration of inflammatory cells into the skin were determined. RESULTS: MTX inhibited random migration and chemotactic migration of PMNs to PAF, LTB4 and PAF-induced intraepidermal accumulation of inflammatory cells in dose- and time dependent fashion. BN52021 specially inhibited PAF-induced chemotaxis of PMNs and intraepidermal accumulation of inflammatory cells, but did not inhibit PMNs random migration and LTB4-induced chemotaxis of PMNs. CONCLUSIONS: The inhibition of PMNs activities may be part of the mechanism of MTX therapy for psoriasis; BN52021 is a selective inhibitor of PAF-induced chemotaxis of PMNs, and therefore can be useful in the treatment of some inflammatory dermatoses such as psoriasis. PMID- 9206082 TI - Diagnosis and treatment of nephrotic syndrome during pregnancy. AB - OBJECTIVE: To set forth the methods of diagnosis and treatment of nephrotic syndrome during pregnancy (NSP) so as to decrease perinatal mortality. MATERIALS AND METHODS: Forty cases of NSP including 5 twins in Tianjin Central Hospital of Obstetrics and Gynecology in the past 13 years have been reviewed, and the diagnosis and treatment were compared between group A (1979-1990, 23 cases) and group B (1991-1992, 17 cases). RESULTS: NSP occurred before the 20th gestational week in 15% of cases and the patient's condition was serious. Clinically, NSP was classified into three types: simple, nephritis and pregnancy induced hypertension (PIH) type. The perinatal maternal and fetal prognosis was poor. Complications occurred in 27.5% of patients. One patient died from eclampsia and disseminated intravascular coagulation (DIC). The total perinatal mortality was 42.22%, with group A (27 babies including 4 twins), 51.86% (14/27), and group B (18 babies including 1 twin), 27.78% (5/18) (P < 0.01). CONCLUSIONS: NSP is a result of reproductive immunology and belongs to type III allergic reaction in terms of immunology. Early diagnosis, early treatment and timely termination of pregnancy may improve the outcome. PMID- 9206083 TI - Age-of-onset related HLA-DQA1 genetic heterogeneity of insulin-dependent diabetes mellitus. AB - OBJECTIVE: To clarify whether there is an association of insulin-dependent diabetes mellitus (IDDM) susceptibility with HLA-DQA1 locus in adult-onset IDDM. PATIENTS AND METHODS: Possible heterogeneity in immunogenetic aspects was investigated by comparing the findings on relationship between HLA-DQA1 52 Arg(+) and IDDM predisposition in three groups with different ages of IDDM onset (group A 14 years, group B 15-30 years, group C > 31 years). RESULTS: The frequency of HLA-DQA1 52 Arg(+) in group A (87.5%) was higher than that in control (53.9%, P < 0.05) and in group B and C (P < 0.05). The frequency of HLA-DQA1 52 Arg(+)/Arg(+) phenotype in group A (75%) was higher than that in group C (23.1%, P < 0.05). CONCLUSION: The contribution of HLA-DQA1 52 arginine to IDDM susceptibility is heterogeneous. Further study is necessary to clarify the immunogenetic entity of IDDM. PMID- 9206084 TI - Antitumor activity of thevetoside in vitro. AB - OBJECTIVE: To study the effects of thevetoside (TS), a cardiac glycoside, and an inhibitor of Na+, K(+)-ATPase, on tumor cells cultured in vitro. METHODS: The cytotoxic effects of TS on tumor cells were determined by trypan blue dye exclusion, neutral red vital staining and clonogenic assay. The time-effect relationship and growth inhibition of tumor cells by TS were assayed with trypan blue exclusion method. RESULTS: TS at low doses (0.005-0.1 mg.L-1), with dose dependence, was able to kill SMMC-7721, SGC-7901 and HeLa cells. IC50 values for SMMC-7721, SGC-7901 and HeLa cells were 0.007, 0.011 and 0.018 mg.L-1 by trypan blue dye exclusion test and 0.016, 0.055 and 0.078 mg.L-1 by neutral red vital staining test. TS inhibited the clonal forming rate of SMMC-7721 and SGC-7901 significantly with IC50 values of 0.021 and 0.036 mg.L-1, respectively. Only when the cells were continuously treated with TS for more than 8 hours, the drug induced cell lethality could be displayed and strengthened quickly. The growth of tumor cells was notably inhibited after they were exposed to 0.1 microgram/ml of TS for 12 hours. All the experimental results of antitumor activity in vitro showed that SMMC-7721 was most sensitive to TS among the three kinds of tumor cells. CONCLUSIONS: TS has cytotoxic action on tumor cells cultured in vitro and this lethal effect must have an action process, in which tumor cells are not dead but suffer from deadly injury and lost the capability of unlimited proliferation. PMID- 9206085 TI - Effect of acupuncture on cardiopulmonary function. AB - OBJECTIVE: To evaluate the relationship between acupuncture and cardiopulmonary function in healthy people. SUBJECTS AND METHODS: Healthy male volunteers were divided into 3 groups, 16 of persons each Group 1 was treated with acupuncture at points Neiguan (PC6) and Zusanli (ST36): Group 2 was treated with acupuncture at non-acupoints, and Group 3 was taken as control. The effects of resting cardiopulmonary functions were measured with gas analysis system. The resting heart rate (HR), oxygen consumption (VO2), and carbon dioxide production (VCO2) were recorded over a thirty-minute period at intervals of five minutes, 15 minutes, and 25 minutes. Electroacupuncture was given to Groups 1 and 2. The analysis of variance and t test were used in data analysis. RESULTS: In the acupuncture groups, resting heart rate and carbon dioxide production decreased (P < 0.05) and oxygen consumption also decreased slightly, although it was statistically insignificant (P > 0.05). CONCLUSION: Our results indicate that acupuncture can decrease the resting heart rate and carbon dioxide production, thus lowering the metabolic rate. PMID- 9206086 TI - Immunohistochemical detection and significance of hepatitis C virus antigen. AB - OBJECTIVE: To study the expression and clinical significance of hepatitis C virus (HCV) antigens in liver tissues of patients with liver disease. PATIENTS AND METHODS: Hepatitis C virus antigen in paraffin-embedded liver tissues was detected by immunohistochemical method using polyclonal anti-HCV and monoclonal anti-HCV NS3. RESULTS: Among the 252 liver tissues, 25 (9.92%) were positive for HCAg (detected with polyclonal anti-HCV), and 19 (7.74%) were positive for HCV NS3 (detected with monoclonal anti-HCV-NS3). All of the 19 HCV NS3 positive tissues were also positive for HCAg. Their distribution in liver tissue was similar, locating in the cytoplasm of liver cells. In most tissues, the positive cells scattered in liver lobulus, without obvious approach to the necrosis and infiltration aggregation; while a few samples showed positive cells clustered or diffused, with some necrosis or/and inflammatory infiltration. Of the 252 sera samples, 71 had HCV antibody or/and HCV RNA, showing a good correlation between HCV antigen in liver tissues and HCV marker in sera, but the detection rate of HCV antigen in liver tissues was much lower than that of HCV markers in sera. Two cases had HCV antigen in their liver with no HCV markers in their sera. CONCLUSION: Stored paraffin-embedded liver tissue can be used in the study of the immunopathology of HCV: no definite relationship between the expression of HCV antigens and liver damage has been found. Further study of the pathogenesis of HCV is needed. PMID- 9206087 TI - CT evaluation of symptomatic ossification of the ligamentum flavum in thoracic spine. PMID- 9206088 TI - Quantitative analysis of the shape of breast mass by ultrasonography: deformity index. PMID- 9206089 TI - Rapid detection of cholera toxin gene of Vibrio cholerae O1 by polymerase chain reaction. PMID- 9206090 TI - Pulsatile secretion of hormones: significance and a tentative idea of future research. PMID- 9206091 TI - Military medical sciences being faced with the 21st century. PMID- 9206092 TI - National survey on catheter ablation. PMID- 9206093 TI - Progress of surgical treatment of intrahepatic lithiasis in China. PMID- 9206094 TI - Acute lower respiratory tract infections in children in China. PMID- 9206095 TI - Pulmonary diseases attract attention in China. PMID- 9206097 TI - Gallium-67 scanning for detection of alveolitis in idiopathic pulmonary fibrosis and sarcoidosis. AB - OBJECTIVE: To investigate 67Gallium scanning for the detection of alveolitis in idiopathic pulmonary fibrosis and sarcoidosis. PATIENTS AND METHODS: 67Ga scintigraphy was performed in 14 patients with IPF (age: mean +/- s = 58.3 +/- 10.3 years), 13 patients with sarcoidosis (age: mean +/- s = 46.9 +/- 9.0 years) and 11 controls (age: mean +/- s = 44.0 +/- 10.5 years). RESULTS: Lung/thigh ratio was respectively 3.18 +/- 0.07 in IPF, 3.12 +/- 0.94 in sarcoidosis and 2.11 +/- 0.26 in controls. IPF and sarcoidosis groups had a significantly higher lung/thigh ratio comparing with the control group (P < 0.05). Three patients received 67Ga scanning examination after corticosteroid therapy and the 67Ga uptake decreased. CONCLUSIONS: The 67Ga scintigraphy is useful in monitoring the activity and extent of alveolitis. PMID- 9206096 TI - The role of human leukocyte antigen in susceptibility and clinical manifestations of sarcoidosis. AB - OBJECTIVE: To investigate the correlation of human leukocyte antigen (HLA) with susceptibility and clinical manifestations of sarcoidosis. MATERIAL AND METHODS: Fifty-five patients with sarcoidosis and one hundred and six normal subjects were investigated. Genomic DNAs were purified using the proteinase K-phenol extraction method. DNA samples were amplified by the polymerase chain reaction (PCR) procedure using the DRB1 group specific primer pairs. RESULTS: The gene frequency of HLA-DR5 increased significantly in patients with sarcoidosis (P < 0.01), and HLA-DR7 decreased (P < 0.05). Gene frequencies of HLA-DR9 and HLA-DR5 increased remarkably in male patients and in patients with stage I and stage IIa, respectively (both P < 0.05). CONCLUSIONS: It is suggested that HLA-DR gene might contribute to the susceptibility as well as various clinical manifestations of sarcoidosis. PMID- 9206098 TI - Protective effect of interleukin-1 receptor antagonist on oleic acid-induced lung injury. AB - OBJECTIVE: To observe the changes of interleukin-1 (IL-1), nitric oxide (NO) and nitric oxide synthase (NOS) in mice with oleic acid-induced acute lung injury (ALI) and the protective effects of interleukin-1 receptor antagonist (IL-1ra). MATERIAL AND METHODS: Male Kunming mice were divided into control, oleic acid and IL-1ra groups. The control group mice were injected saline; the oleic acid group mice were injected oleic acid (0.2 ml/kg): and the IL-1ra group mice were injected the IL-1ra (20 mg/kg). Lung index, lung wet-to-dry weight ratio, and total protein, cell analysis, nitric oxide measurement, NOS activity in BALF, lung pathology examination were made after an hour of administration of drug. RESULTS: Preadministration of IL-1ra to the mouse with ALI decreased the lung index, lung wet-to-dry weight ratio and leakage of protein from pulmonary capillary, elevated PaO2, and attenuated lung histologic injury. It was found that in bronchoalveolar lavage fluid (BALF), NO amount and lung NOS activity increased in oleic acid group, BALF NO amount and lung NOS activity decreased obviously after given IL-1ra. CONCLUSIONS: This study demonstrated the protective effect of IL-1ra on oleic acid-induced lung injury, NO may participate in the pathological process of lung injury. PMID- 9206099 TI - Steroid-resistant asthma immunologic characteristics. AB - OBJECTIVE: To investigate whether persistent T lymphocyte activation is a feature of steroid-resistant (SR) asthma and to study the inhibitory effects of dexamethasone and other immuno-inhibiting agents on PHA-induced proliferation of peripheral blood T lymphocytes from SR and steroid-sensitive (SS) asthmatics. MATERIAL AND METHODS: 15 SR and 15 SS asthmatics were studied. Serum levels of soluble interleukin-2 receptor (sIL-2R) were measured before and after prednisone therapy by an enzyme-linked immunosorbent assay. PHA-driven proliferative assay was performed to evaluate inhibition of T lymphocyte proliferation. RESULTS: Serum levels of sIL-2R were elevated in both patients with SR and SS asthma as compared with normal controls (P < 0.001). After a 7-day course of prednisone (20 mg/day), serum levels of sIL-2R decreased significantly in SS asthmatics (P < 0.001) but not in SR asthmatics (P > 0.1). Proliferation of T lymphocytes from the sensitive but not the resistant asthmatics was significantly (P < 0.002) inhibited by dexamethasone (10 mol/L), reflecting a shift of the dose-response curve. In contrast, oxymatrine and thymus-derived immunosuppressors inhibited proliferation of T lymphocytes to a similar degree between SR and SS asthmatics. CONCLUSIONS: The results suggest that persistent T lymphocyte activation due to a relative insensitivity of the cells to glucocorticoids is a feature of SR asthma. Immuno-inhibiting agents other than glucocorticoids may be of therapeutic benefit in patients with SR asthma. PMID- 9206100 TI - Expression of TGF-beta 1, PDGF and IGF-1 mRNA in lung of bleomycin-A5-induced pulmonary fibrosis in rats. AB - OBJECTIVE: To investigate the influence of alveolar macrophages (AMs), fibroblasts and interstitial cells on development of lung fibrosis, and the interactions among TGF-beta 1 PDGF and IGF-1 and these cytokines-effects on lung fibrosis. MATERIAL AND METHODS: Expressions of TGF-beta 1, PDGF and IGF-1 mRNA in the lung cells and lung tissues in different stages of Bleomycin-A5-induced pulmonary fibrosis in rats were studied through Northern hybridization. RESULTS: The expressions of TGF-beta 1 and PDGF mRNA reached their peaks in AMs of pulmonary fibrosis in rats on the 7th day after Bleomycin-A5 instillation. It was similar with that in the lung tissues. IGF-1 mRNA remained relatively stable in AMs during the course. PDGF and IGF-1 mRNA increased gradually in fibroblasts, and reached the highest expressions in the interstitial cells. There was almost no TGF-beta 1 mRNA expression in all groups of fibroblasts. CONCLUSIONS: AMs are the main sources of TGF-beta 1 and PDGF in the lung tissues with fibrosis induced by Bleomycin-A5 AMs are activated in the first weekend and secrete TGF-beta 1 and PDGF to promote fibroblasts proliferation and fibrosis. As fibrosis developed, fibroblasts have established PDGF and IGF-1 autocrine and these three cytokines paracrine nets combined with the interstitial cells to promote lung fibrosis. PMID- 9206101 TI - Spontaneous apoptosis in human colon tumor cell lines and the relation of wt p53 to apoptosis. AB - OBJECTIVE: To examine spontaneous apoptosis of cultured human colon tumor cell lines in vitro and to investigate the role of wild type (wt) p53 in regulation of apoptosis induced by DNA-damaging treatment. METHODS: A model system of human tumor progression involving three cell lines was used in this study for examination of apoptosis. They were originally established from human colon villous adenoma, including an early passage of non-tumorigenic cell line, V235E; a late passage of weakly tumorigenic cell line, V235L; and a spontaneous progressing highly tumorigenic cell line. V411. All of them maintain wt p53 expression. For identification of apoptosis, two tests were performed: 1. morphology study using acridine orange (AO)/ethidium bromide (EB) stainning by fluorescence microscopy; 2. DNA electrophoresis on agarose gel. P53 and WAF-1 (a downstream gene of p53) expressions were analysed at mRNA level using Northern blot technique. Apoptotic index of cell lines examined was measured by DNA fluorescence assay. RESULTS: Spontaneous apoptosis was demonstrated in cell lines of all stages of progression by both morphology and DNA agarose gel electrophoresis. Apoptosis was further induced in V411 after treatment of cells with 137Cs gamma-irradiation and accompanied by increases in p53 and WAF-1 expression. In contrast, a mutant p53 bearing human colon cancer cell line, sw480, lacked spontaneous apoptosis, and upon irradiation neither induction of apoptosis nor increase expression of p53 and WAF-1 were seen. CONCLUSIONS: Apoptosis can be maintained in some human tumor cell lines despite transformation and carcinogenesis. Wt p53 and WAF-1 products are two of the potential mediators which effect apoptosis. Additionally, since apoptosis was enhanced by irradiation in V411, but not in sw480, it suggests that wt p53 cancer cells are more sensitive to DNA-damaging treatment than mutant p53 cancer cells. These finding may have implications for cancer therapy. PMID- 9206102 TI - Observation on solubility and pro-nucleative activity of monoconjugated bilirubin. AB - OBJECTIVE: To explore the effect of monoconjugated bilirubin (MCB) on gallstone formation of both categories. METHODS: Using a newly established high performance liquid chromatography (HPLC) technique. MCB was extracted, prepared and purified from gallstone patients bile, and its solubility and pro-nucleative activity were observed. RESULTS: In a near physiological environment of biliary tract (pH 7.9 and temperature 37 degrees C), the aqueous solubility of MCB (558.25 +/- 5.96 microns) was only 1/7 of that of disconjugated bilirubin (DCB) (3410.9 +/- 2.42 microns), but 44 times higher than that of unconjugated bilirubin (UCB) (2.43 +/- 0.31 microns) (P < 0.01, between and two of them). Moreover, the solubility of MCB changed curvilineally with the alteration of pH, being highest at pH 7.9, zero at pH 4.5 and even lower than that of ionized UCB at pH above 9. In 3 groups of model bile systems made from Kibe's formula, the nucleation time (NT) was examined by polarized microscopy after 170 microns and 340 microns of MCB was added in group 1 and 2 respectively (group 3 was the control, with no MCB added). The NT was 3.33 +/- 0.52. 2.17 +/- 0.41 and 5.83 +/- 0.75 days respectively in the 3 groups (P < 0.05, between any 2 of the 3 groups) and the nucleative activity (NA) was 0.57 and 0.37 in group 1 and 2 respectively, which confirmed that MCB possessed a dose-related NA which was even stronger than that of glycoprotein (0.56) at the dose of 340 microns. CONCLUSIONS: MCB might play an essential role in the formation and nucleation of gallstones with its poor aqueous solubility and strong pro-nucleative activity via certain lithogenic mechanism. PMID- 9206103 TI - Neurotrophic effects of Schwann cells in culture on spinal anterior horn neurons. AB - OBJECTIVE: To study the neurotrophic effects of Schwann cells in culture on damaged spinal anterior horn neurons. MATERIALS AND METHODS: Dissociated from rat sciatic nerves undergoing Wallerian degeneration by trypsin-collagenase. Schwann cells were cultured for 96 hours to form a trophoblast, onto which embryonic day 14 rat spinal anterior horn neurons plated on glass coverslip and cultured for 4 hours were transferred for cocultivation without cell-cell contact between Schwann cells and spinal anterior horn neurons. The effect of survival and neurite outgrowth activity promoting factors secreted from Schwann cells on spinal anterior horn neurons were examined under phase-contrast microscope at 24, 48, 72 and 96 hours after cocultivation. RESULTS: When cultured for 24 hours, almost all the spinal anterior horn neurons, that had been damaged in the course of dissociation from spinal nervous tubes, survived, and some of them grew with short neurites. 96 hours after cocultivation, about 1/3-1/2 of the neurons survived and their bodies enlarged with neurites by more than six times of their body diameters. Contrarily, none of the neurons in the control group (no Schwann cell trophoblast) survived simultaneously. CONCLUSIONS: By the model of Schwann cell-spinal anterior horn neuron cocultivation separated by glass coverslip, it was well confirmed that Schwann cell can secrete a neuron survival promoting factor and a neurite outgrowth promoting factor, which undoubtedly play a crucial role in peripheral nerve regeneration. PMID- 9206104 TI - Guided bone regeneration in long bone. An experimental study. AB - OBJECTIVE: In the concept of guided tissue regeneration (GTR), a space is created for selected cells to differentiate, proliferate and repair the defect at last by implanting a membrane around the defect area, which serves as a physiologic barrier. This experiment was designed to test GTR in long bone, namely, guided bone regeneration (GBR). METHODS: Ten New Zealand rabbits were used in this experiment. A piece of silicone membrane sutured as a tube was used to bridge a 10-mm defect on radius. 10-mm defects were also produced on the control sides. Radiography of forelimbs was taken weekly until 12 weeks. Gross sample examination, 3-point bending test and histology were involved in evaluating bone regeneration. RESULTS: By the 12th week, seven of 10 experimental sides were healed, 2 were healed with a connective cartilage zone, and I was not healed. None of the control was healed but the defect was occupied by soft tissue. CONCLUSIONS: The results of this preliminary study showed that GBR is present in long bone of rabbits. The following points relate to the mechanism of GBR; providing a space for bone regeneration; preventing surrounding tissue from the defect; increasing the density of osteogenic precursor cells and concentration of bone morphogenetic protein (BMPs); and maintaining a complete blood clot to bridge the fracture ends in the tube, which provides a structure for osteogenic cells ingrowth. PMID- 9206105 TI - Chronic suppurative parotitis: a proposed classification. AB - OBJECTIVE: To propose a practical and reasonable classification of chronic suppurative parotitis (CSP) on the basis of the various entities. MATERIAL AND METHODS: Clinical, laboratory, sialographic, scintigraphic, histopathologic (including ultrastructural) study of recurrent parotid swellings (RPS) was performed in 291 patients over a 10-year period. RESULTS: It is suggested that CSP should be classified into recurrent parotitis in childhood (RPC), recurrent parotitis in adults (RPA), chronic obstructive parotitis (COP) and should be differentiated from other subdivisions including subclinical Sjogren's syndrome (SCSS), chronic parotid swelling of Sjogren's syndrome and sialadenosis with retrograde infection. RPA is a continuation of recurrent parotid swelling from childhood (RPC) to adulthood. Remission can take place spontaneously in RPC and RPA so that self-conservative therapy is mainly used for reducing the parotid swellings. COP is recurrent parotid swellings and/or purulent discharge resulting from various obstructive factors. Mild COP can recover completely with the use of conservative methods, severe COP is often resistant to conservative treatment and should be treated with surgical modality or injection of methyl violet into the diseased gland. Treatment with methyl violet is considered as a convenient and practical method with a definite effect. SCSS is an autoimmune disease and should be treated as systemic disease. CONCLUSIONS: Because there exists confusion in the nomenclature of RPS this revised classification is based on the various entities and can be used as a guide in the diagnosis and treatment of RPS. PMID- 9206106 TI - Cataract free zone and primary health care approach to prevention of blindness in Shunyi county of Beijing. AB - OBJECTIVE: To establish a three-level eye care network, together with a referral and monitor system for the prevention of blindness, with the aim of creating a cataract free zone. SUBJECTS AND METHODS: Shunyi County with a population of 548364 was chosen as the target site. Our activities included (1) establishment of a County Guiding Committee for the prevention of blindness: (2) training local ophthalmologists and primary eye care personnel: (3) extensive publicity of knowledge for the prevention of blindness: (4) screening for blindness and low vision, (5) cataract surgery as the chief measure for creating a cataract free zone. RESULTS: A referral and monitor system for cataract blindness was established in 1987. Seven local ophthalmologists, 449 primary eye care health workers and six optometrists were trained. 815 cataract-blind patients were identified. Up to December 1991, 667 patients (737 eyes), accounting for 81.84% of the total, were treated with cataract surgery, 90.64% of these eyes had their sight restored, and 59.66% patients resumed work. A cataract free zone has been created in Shunyi County. CONCLUSIONS: The prevention of blindness with cataract surgery as the chief measure can dramatically decrease the prevalence of blindness in a relatively short period of time. PMID- 9206107 TI - Searching eye movement, smooth pursuit eye movement and schizophrenia. AB - OBJECTIVE: To detect whether the smooth pursuit eye movement (SPEM) and searching eye movement (SEM) could be considered as a biological marker of schizophrenia, and used as a tool in helping diagnosis of schizophrenia. METHODS: 88 schizophrenics, 77 patients with mood disorders, 32 with "neurosis", and 74 normal healthy controls were examined for SPEM and SEM individually. The authors verified the results in all the first-visit 150 outpatients in March 1993 by comparing the examination results with the clinical diagnoses after a 6-month follow-up. RESULTS: Significant differences were found in the number of eye fixation (NEF) and total eye scanning length (TESL) of SEM between schizophrenics and normal controls or patients with other disorders. Less NEF and shorter TESL could be helpful in differential diagnosis, and the agreement rate, Kappa coefficient was 0.62. No significant differences were found in SPEM in this investigation between non-medicated schizophrenics and normal controls. CONCLUSION: Searching eye movement (SEM) might be considered as a biological marker of schizophrenia and might be used as a supplementary tool in its diagnosis. PMID- 9206108 TI - Relations between the sides of linguistic cerebral dominance and manuality in Chinese aphasics. AB - OBJECTIVE: To study the relations between the sides of linguistic cerebral dominance and of cerebral lesions and manuality in Chinese aphasics and whether there is essential difference between Chinese Han nationality and Westerners in the linguistic cerebral dominant laterality. METHODS: 309 cases of cerebral infarction (225 cases) and cerebral hemorrhage (84 cases) at the acute stage (within 3 months) of illness were studied by Standardized Aphasia Battery in Chinese (ABC) and Manuality Test (12 items). All cases were at their first onset of cerebral vascular disease (CVD) and were orientated. Cranial CT scan demonstrated single cerebral lesion in each case. Aphasia was divided into 10 types, and manuality was divided into dextrals and nondextrals. RESULTS: 286 cases (92.56%) were dextrals and 23 cases (7.44%) were nondextrals. The lesions of 222 cases (71.85%) were in the left cerebral hemisphere and 87 cases (28.15%) were in the right cerebral hemisphere. 170 cases (55.02%) were normal in their speech and 139 (44.98%) with aphasia of 10 types, of whom, 134 cases (96.40%) were dextral and 5 (3.60%) were nondextral. 136 patients (97.84%) were aphasic due to lesions of the left cerebral hemisphere, of whom, 131 cases (94.24%) were dextral and 5 (3.60%) were nondextral. Only 3 dextrals (2.16%) were with crossed aphasia due to lesions of the right cerebral hemisphere. CONCLUSIONS: The majority of the Chinese Han nationality had their linguistic cerebral dominance in the left cerebral hemisphere, no matter whether they were dextral or not, which had no essential difference between Chinese and Westerners. PMID- 9206109 TI - The central distribution of adrenomedullin and its effects on blood pressure and heart rate in rats. AB - The present study was designed to make certain whether there exists adrenomedullin (ADM) in the rat central nervous system and evaluated the hemodynamic actions of intracerebroventricular administration (ICVA) of human ADM[13-52]. By immunohistochemistry (ABC method). We found that there was a discrete localization of ADM-positive immunoreactivity in the rat central system including cerebral cortex, paraventricular tissues, hypothalamus, cerebella cortex, mesencephalon and medulla oblongata. By reverse transcription-polymerase chain reaction (RT-PCR) analysis, rat ADM mRNA was found to be expressed in rat brain. These above results of immunohistochemistry and RT-PCR suggest that ADM exists in the rat brain. We also found that centrally administered ADM[13-52] in a dose of 0.4 to 3.2 nmol/kg provoked marked, prolonged and dose-dependent increases in mean arterial blood pressure (MABP) and heart rate (HR). To clarify the mechanisms of the hemodynamic changes induced by centrally administered ADM [13-52], the effect of centrally administered ADM[13-52] on renal sympathetic nerve activity (RSNA) was studied. The result showed that centrally administered ADM [13-52] (1.6 nmol/kg) provoked a marked increase in RSNA, therefore, the increases in MABP and HR induced by centrally administered ADM [13-52] might be due to the stimulation of central sympathetic mechanism. In addition, we also compared the relationship of activity and structure among the different fragments of ADM. In conclusion, ADM exists in the rat brain, and it may play an important role in the central control of cardiovascular system. PMID- 9206110 TI - Re-evaluation of the mechanism and treatment of angina decubitus. AB - 30 patients with angina decubitus (AD) were studied during hospitalization. These patients were found to have severe coronary artery obstructive lesions and an increase of myocardial oxygen consumption (MOC) before the onset to AD, indicating that AD belongs to the category of effort angina. 18 patients were investigated by continuous hemodynamic monitoring. Three patients had significant increase in pulmonary artery diastolic pressure (PADP) before the onset. In the other 15 patients, PADP increased slightly in 12 and remained unchanged in 3 cases before the onset. Left ventriculography showed ejection fraction (EF) > 45% in 25 of the 27 patients. These results indicate that left ventricular (LV) systolic dysfunction is not a major factor in the pathogenesis of AD. The patients with LVEDP > 12 mmHg constituted 60% of 25 patients with EF > 45%, suggesting that these patients had obvious LV diastolic dysfunction, which may be the major factor in the pathogenesis of AD. According to the results of our treatment, beta blockers may be used as the major form of treatment in the patients with AD. PMID- 9206111 TI - Reduced tumorigenicity of metastatic human lung cancer cell subline (PGCL3) transfected with hRAR beta gene. AB - The recombinant PSG5-RAR beta plasmid and the G418-resistant PSV2 neo plasmid (10.1) were cotransfected into PGCL3 cells by coprecipitation with calcium phosphate. The transfectants CR3 and CR4, which expressed the RAR beta gene, were identified by Northern blot hybridization. The results showed that the in vitro growth and invasion of CR3 and CR4 were dramatically reduced compared to the control-transfected cell (CSV1). Furthermore, the colony-forming abilities in soft agar and the tumorigenicity in nude mice of CR3 and CR4 were abrogated. Our results suggests that RAR beta functions not only as a receptor mediating the RA action, but also as a suppressor in lung tumorigenesis. PMID- 9206112 TI - The amplification and expression of MDR1 gene in adriamycine resistant cell line of colon cancer cell HR8348. AB - P-glycoprotein plays an important role in highly drug resistant cells. But its high expression cannot be achieved by chemotherapy. In order to study the effect of P-glycoprotein on clinical tumors, we established a low ADM resistant colon cancer cell line HR/ADM and determined the amplification and expression of mdr-1 gene. The GLC/ADM showed a resistant pattern similar to classical MDR and the transcription of mdr-1 gene determined by RT-PCR increased. The immunocytochemical analysis showed strong positive staining with monoclonal antibody. The gene amplification of mdr-1 was clearly demonstrated by southern blot. Our results suggested that moderate expression of P-glycoprotein might be enough for a high resistant pattern. PMID- 9206113 TI - Relaxant effects of vasoactive intestinal peptide on pulmonary artery in chronically hypoxic rats. AB - The object of this study is to investigate the effect of VIP on pulmonary artery of chronically hypoxic rats. It was shown that chronic hypoxia depressed significantly pulmonary artery relaxation induced by VIP as compared with those of control (P < 0.001). The vascular relaxation of both groups was correlated with concentration of VIP. In addition, the relaxant effect of VIP on pulmonary arteries in rats was endothelium-independent, and was not prevented by indomethacin or nordihydroguaiaretic acid, but was abolished completely by methylene blue. These results suggest that the lower relaxation of pulmonary artery in rats might not be due to the endothelial injury caused by chronic hypoxia, and chronic hypoxia may inhibit directly the soluble guanylate cyclase in vascular smooth muscle cells involved in synthesis of cGMP and thus reduced the sensitivity and reactivity of pulmonary artery to VIP. PMID- 9206114 TI - Effects of alveolar macrophage conditioned media from interstitial lung disease patients on the procollagen mRNA expression in human lung fibroblasts. AB - Progressive inflammation and fibrosis are the central processes in the pathogenesis of pulmonary fibrosis. It is believed that macrophages in areas of chronically inflamed lung play a key role in fibrotic response. Therefore, we investigated the effects of alveolar macrophage (Am phi) conditioned media from interstitial lung disease (ILD) patients on lung fibroblast proliferation and procollagen mRNA expression. After stimulating with Am phi conditioned media from ILD patients, the fibroblast proliferation increased 71.4% compared with the control, but for media from bronchial carcinoma (BC) patients, it just increased 14.3%. There is a significant difference between the two groups (P < 0.05). The procollagen alpha, (I) mRNA in fibroblasts stimulated with Am phi conditioned media from ILD patients was increased 21.3%, and alpha 1 (III) was 37.2% higher than control (P < 0.05). It increased 6.8% and 12.8% for media from BC patients respectively, but there was no difference when compared to the control. We considered that Am phi from ILD patients might be in an activated state and could release some growth factors to stimulate fibroblast proliferation and promote collagen DNA expression. PMID- 9206116 TI - Intrapancreatic cholecystokinin mediates vagally stimulated exocrine secretion from the rat pancreas. AB - Although cholecystokinin is localized within neuronal fibres of the pancreas, a physiological role for intrapancreatic cholecystokinin has not been identified. The strategy of this study was to elicit pure vagal stimulation electrically, and to use specific receptor antagonists to identify the mediators of exocrine pancreatic secretion. We conclude that vagal stimulation of the rat pancreas involves ganglionic neurotransmission and release of acetylcholine and cholecystokinin from intrapancreatic, postganglionic fibres. To our knowledge, this is the first study to demonstrate a physiological role for intrapancreatic cholecystokinin. PMID- 9206115 TI - Hypoxia and endothelin-1 stimulate DNA synthesis of pulmonary artery smooth muscle cells. AB - Hypoxia and endothelin-1 (ET-1) are associated with constriction of pulmonary vasculature both in vivo and in vitro. However, the role of hypoxia and ET-1 in the vascular remodelling during the development of pulmonary hypertension is unclear. This study demonstrated that ET-1 (0.1 nmol/L to 100 nmol/L) increased the [3H] thymidine uptake in a dose-dependent manner in cultured bovine pulmonary artery smooth muscle cells (PASMC), which was enhanced by exposing PASMC to hypoxia (2% O2, 93% N2, 5% CO2). BQ123, the specific antagonist of endothelin receptor subtype A, eliminated the ET-1 medicated proliferation of PASMC and the cooperative effect of hypoxia. Some dilatory drugs could inhibit the mitogenic effect of ET-1. We also observed that hypoxia significantly increased [3H]thymidine uptake in PASMC without ET-1 and BQ123 could inhibit this effect. Radioimmunoassay suggested that there was an autocrine of ET-1 in cultured PASMC which was enhanced by hypoxia significantly. PMID- 9206117 TI - Analysis of the N-glycosylation of the serum glycoproteins defined by Con A, PHA E, RCA1 and WGA in Chinese patients with gastrointestinal diseases. AB - Glycoproteins from tumor cells isolated by Con A, PHA-E and RCA1, were coupled to horse radish peroxidase and then used as reporters to evaluate the oligosaccharides change on serum glycoproteins in digestive tract diseases and individual healthy persons. A value 2 standard deviation above the mean of the healthy person group was considered positive. The results indicated that the positive rates of the oligosaccharides bound with Con A, PHA-E and RCA1 were 51% (61/119), 70.6% (84/119), and 76.4% (91/119) in patients with gastric cancer; 53.3% (16/30), 46.7% (14/30) and 66.7% (20/30) in esophageal cancers; 28.5% (15/49), 49.0% (24/49) and 46.9% (23/49) in colorectal cancer; 78.1% (25/32), 81.3% (26/32) and 31.3% (10/32) in gallbladder tumors; 60.4% (29/48), 54.1% (26/48) and 39.5% (19/48) in hepatocellular cancer. The positive rates among the patients with benign diseases were also found to be 12.7% (16/126), 15.5% (20/126) and 26.2% (33/126). However, the expression of the oligosaccharide moieties recognized by WGA on the glycoprotein bound with Con A, was significantly lower in the serum of patients with tumors than in those with benign disease and in healthy individuals (P < 0.01, P < 0.01). PMID- 9206118 TI - Omniplane transesophageal echocardiography imaging planes exploration. AB - One hundred and twenty-four patients with heart disease were examined by omniplane TEE in order to systematically research every views of omniplane TEE, and further explore anatomy and image feature of each view. The result showed that omniplane TEE transducer can be rotated in probe from 0 degree to 180 degrees, obtain many views at various angles behind the heart and fully demonstrate the structure and pathology of the heart and great vessels. It was useful for clinical diagnosis because of getting more information about the heart and great vessels. As omniplane TEE probe was little rotated in esophagus, it lessened esophagus stimulation. Meanwhile, it was suitable for three-dimensional reconstruction of left ventriculum. PMID- 9206119 TI - Effects of systemic fluconazole therapy on in vitro adhesion of Candida albicans to buccal epithelial cells and changes of the cell surface proteins of the epithelial cells. AB - This paper presented the effects of systemic fluconazole therapy via intravenous (IV) and oral (PO) administrations on the adhesion of Candida albicans (C. albicans) to the buccal epithelial cells (BEC) from five treated patients with three candidosis, one mucormycosis and one sporotrichosis and at the same time, an analysis of the cell surface proteins involving candidal adherent receptor in the BEC of the patients in the course of 7 days were exposed to 3H-leucine radiolabeled C. albicans for in vitro candidal adherent assay, and the BEC from first intake day and the last intake day of the patients were extracted by dithiothreitol (DTT)-iodoacetamide treatment for SDS-PAGE. These results indicate that the systemic fluconazole therapy results in the inhibitory effect of candidal adhesion to BEC of treated patients to prevent them from oral candidosis for a prolonged time, which is based on the absent surface protein (35 KDa) of the BEC. PMID- 9206120 TI - Effect of 764-3 on aggregation and calcium movements in aequorin-loaded human platelets. AB - Washed human platelets were loaded with the Ca(2+)-sensitive photoprotein, aequorin, using hypoosmotic shock treatment-technique. Then aggregation and cytoplasmic ionized calcium concentration ([Ca2+]i) changes in response to collagen or thrombin were measured simultaneously in the aequorin-loaded human platelets with a Platelet Ionized Calcium Aggregometer. 764-3, an active component isolated from the Chinese medicinal herb Salvia Miltiorrhiza Bge, inhibited platelet [Ca2+]i rise as well as aggregation evoked by collagen or thrombin in the presence of extracellular Ca2+. After the extracellular Ca2+ was removed by addition of EGTA, collagen or thrombin, causing no aggregation, still elicited platelet [Ca2+]i rise which reflected Ca2+ mobilization from intraplatelet stores. Under this condition, 764-3 could also suppress platelet [Ca2+]i rise. Analysis shows that 764-3 inhibits platelet Ca2+ influx and Ca2+ mobilization with similar potency, which accounts for its suppression of platelet [Ca2+]i rise, and must contribute to its inhibition of platelet aggregation. PMID- 9206121 TI - Differentiation of pseudocondyloma of vulva and condyloma acuminata by dot blot hybridization and polymerase chain reaction. AB - This study differentiated pseudocondyloma of vulva from condyloma acuminata using dot blot hybridization and polymerase chain reaction (PCR). A total of 27 cases of pseudocondyloma of vulva and 65 cases of condyloma acuminata were selected for the study. The genital lesions were examined clinically and were biopsied. Each biopsy was subjected to histological examination and HPV DNA analysis by dot blot hybridization and PCR. Dot blot analysis detected HPV DNA in 19(82.6%) out of 23 cases of condyloma acuminata and 2(25%) out of 8 cases pseudocondyloma of vulvae (P < 0.05). PCR detected HPV DNA in 51(92.7%) out of 55 cases of condyloma acuminata, compared with none in 23 cases of pseudocondyloma (P < 0.001). HPV DNA was present in the majority of condyloma acuminata specimens, HPV 6 and 11 were the predominant types. Peudocondyloma is probably not associated with HPV. PCR was the most sensitive and useful technique for HPV DNA detection. PMID- 9206122 TI - Sexually transmitted diseases: incidence and distribution. AB - The incidence of sexually transmitted diseases (STDs) increased from 26.04 per 100000 in 1987 to 104.81 per 100000 in 1993 in selected areas of the country. Gonorrhea is by far the most common STD but its constituent ratio declined because of a rapid increase of nongonococcal uretheritis and genital warts during most recent years. The incidence of syphilis is relatively low and cases of congenital infection are noted. The wide spread of resistant Neisseria gonorrhoeae infection gives a challenge to the therapeutical and control strategies of STDs. Sexually transmitted Chlamydia trachomatis infections, an important cause of urethritis, cervicitis and pelvic inflammatory disease, is becoming common in our country. Attention has been drawn on viral hepatitis in their means of transmission by sexually behaviors, and also, on the homosexuals, assumed to be the high risk group to catch STDs. Coordinated national efforts to control STDs in China have been taken. PMID- 9206123 TI - [A study of the expression and localization of tumor necrosis factor mRNA in small intestine of rats after severe burn]. AB - In this study, the dot blotting and in situ hybridization, collaborated with determination of contents of endotoxin and TNF in plasma, were used to study the expression and localization of TNF mRNA in the intestine. The results showed that, the expression of TNF mRNA in the small intestine increased rapidly and reached maximum at 6 h postburn, being 5.56 times higher than that of normal control. The TNF mRNA-producing cells in normal animal were mononuclear phagocytes located in lamina propria. After burn, the number of positive cells in lamina propria increased significantly, and the TNF gene-expressing cells were vascular endothelium. Many positive cells were found in the crypts of the intestine. It suggested that the expression of TNF correlated well with bacterial translocation in the early stage of burn. The mononuclear phagocytes and endothelial cells located in lamina propria and interstitial tissue were the chief TNF mRNA-producing cells in the intestine. The overproduction of TNF could provoke microvascular injury or induce the production of oxygen free radical to damage the gut mucosal barrier. PMID- 9206124 TI - [Protective effect of lipid A monoclonal antibody against burn sepsis in rats]. AB - A lipid A monoclonal antibody(mAb) was prepared and it was used to study its protective effect against burn sepsis. Wistar rats were inflicted with 30% TBSA third degress burn, and they were given LPS to mimic early sepsis after burn. The rats were divided randomly into burn with LPS, monoclonal antibody treatment, and control groups. The levels of endotoxin, tumor necrosis factor, light and electron microscopic studies of the morphological changes in the liver were studied. The results showed that the anti-lipid A monoclonal antibody demonstrated capacity to cross-react with several Gram-negative bacteria and their endotoxins. The mAb improved the survival rate of rats and decreased the levels of endotoxin and TNF as well as the liver damage significantly. PMID- 9206125 TI - [Protective effect of ATP-MgCl2 on gut mucosa in burn rat]. AB - In order to study the protective effect of ATP-MgCl2 on gut mucosal barrier function in trauma-induced stressed animal, full thickness burn of 30% total body surface area (TBSA) was produced in rats, and ATP-MgCl2 was injected intraperitoneally right after scald-injury. Malondialdehyde (MDA) contents of ileum mucosa were determined and morphological alteration in the ileum was observed under light microscope. The results showed that ileum mucosal MDA content increased significantly, accompanied by obvious morphological change, after the injury, whereas MDA content was maintained at the normal control level and pathological alteration in ileum tissue was abated after the use of ATP MgCl2. It suggests that ATP-MgCl2 possesses a protective effect on gut mucosal barrier function of scald-injured rat, and the mechanism involved may be related to the inhibition of lipid peroxidation in gut mucosa. PMID- 9206126 TI - [Effects of thermal injury on heart sialic acid content in rats]. AB - The myocardial sialic acid content, ATPase activities, and Ca2+ level were investigated in rats with full thickness burn of 30% TBSA. The results showed that the burned rats had a decreased level of myocardial sialic acid which was only 58.8% of controls at 3 h postburn. Myocardial Na+, K(+)-ATPase, Ca2+, Mg(2+) ATPase and Ca(2+)-ATPase activities were inhibited in burned rats. As compared to controls, the burned rats showed a higher level of Ca2+ in heart tissue. This study indicated that there were abnormal energy metabolism, dysfunction of ion pump, and paradox Ca2+ overload in burned rats, which may be associated with the decrease in sialic acid content in myocardium. PMID- 9206127 TI - [Changes in intracellular K+ concentration and ATPase activity of myocardiocytes in early stage of burn injury]. AB - In this experiment, intracellular K+ concentration ([K+]i) and ATPase activity of myocardiocytes were measured in early stage of burn injury. Comparing with control group, it was found that, 1. [K+]i were decreased after burn injury, [K+]i of 1st, 3rd, 8th and 24th hours were decreased to 96.2 +/- 1.3%, 85.8 +/- 1.3%, 65.9 +/- 1.0% and 73.7 +/- 1.1% of normal, respectively. 2. Cardiac sarcolemma total ATPase, Mg(2+)-ATPase and Na(+)-K(+)-ATPase activities were all reduced significantly at 8th hour after injury. These results suggest that, burn injury accelerates K+ efflux current, but inhibits K+ influx current, and the reduction of Na(+)-K(+)-ATPase activity is one reason of decrease of [K+]i after injury. PMID- 9206128 TI - [Early changes in cardiac function and beta-ARs in heart after severe scalds in rat]. AB - The changes in left ventricular function and ventricle beta-adrenoceptors (beta ARs) were measured in rat undergoing a full thickness scalded burn of skin of 30% total body surface area. The results showed that there were a significant decrease in left ventricular systolic pressure (LVSP), maximum rising and falling rate of left intraventricular pressure (LV +/- dp/dtmax) and density of ventricular beta-ARs. Correlation analysis showed that the change in beta-ARs density correlated with the change in LVSP, LV +/- dp/dtmax positively. PMID- 9206129 TI - [Experimental study on delayed effect of external skin expansion]. AB - In using the external skin expander, we found that the expanded skin had the feature similar to a delayed flap. To study its mechanism, we chose 20 New Zealand rabbits and divided them into 2 groups randomly. The skin on the animal's back in the experimental group was expanded with the external skin expander. The laser Doppler flowmeter (LDF) and oxyhemograph (SpO2) were applied to detect skin microcirculation. The survival area and the results of microangiography of the two groups were compared. Results showed that the value of LDF of the expanded skin increased and the value of SpO2 returned to its origingal level at 12 days after expansion. The survival area in the experimental group was larger than the control. Microangiographic examinations indicated that the subcutaneous vascular anastomoses were obviously increased in the experimental group. We conclude that external skin expansion does not affect the quality of the expanded skin, and its effect is similar to delay technique. PMID- 9206130 TI - [The application of expanded delto-pectoral flaps in the faciocervical region]. AB - The delto-pectoral flap has been used to repair burned scar in the faciocervical region for some years. However, its limited size restricts its application. Furthermore, direct transfer of the flap may result in a swelling and inexpressive face and the donor site needs skin grafting. To avoid the above disadvantages, we have tried pre-expansion of the flap. In this article, the authors report the experiences in the application of the method to eighteen cases, including the surgical procedure, the applied anatomy, and typical cases as well. Also included in the article are the comparison between various therapies to the burned scar of the face, the key points for successful pre expansion of the delto-pectoral flap, accurately maintaining the desired position. The method has been proved to have many advantages and can be widely applied. PMID- 9206131 TI - [Clinical application of 72-hour fast expansion of the soft tissue]. AB - The rapid expansion of the soft tissue has been applied in plastic surgery. To further shorten the expansion period, we undertook a clinical study on 72-hour fast expansion and applied this technique to 11 patients with different defects on their face, neck, shoulder or elbow after resection of scar, melanoma, angioma or due to trauma. Satisfactory results have gained. This paper not only introduces the clinical application of the technique, but also discusses the theoretical basis, advantages and key points of the operation. PMID- 9206133 TI - [Treatment of the deformed ear and jaw]. AB - Lengthening of the mandible by gradual distraction and ear reconstruction using tissue expanders were performed on nine young patients (6-11 years old) with unilateral hemifacial microsomia. The technique holds promise for reconstruction of ear and mandible at the same time. This is essentially a pilot clinical project. A late follow-up of these patients is necessary. PMID- 9206132 TI - [The development of a skin-stretching device and its clinical application]. AB - A skin-stretching device that is designed to harness the elastic properties of the skin using incremental traction is presented. In using this device, the skin is stretched from its edges by direct linear traction. The device produces extra skin tissue rapidly, and no undermining is necessary to loosen skin, so as to close large areas of skin shortage that would otherwise have required more complicated procedures for their repair. It can be employed for a duration of 20 640 minutes, and be applied preoperatively, intraoperatively or postoperatively. 52 patients have undergone the treatment since March 1994. Six months to one year's follow-up of 27 cases demonstrated satisfactory results. The method has following advantages: easy in manipulation, saving time, safe and fewer complications. PMID- 9206134 TI - [Repair of the over-convex antihelix]. AB - Contrary to a prominent ear, the deformity of the over-convex antihelix shows a very acute antihelix angle and helix depression and retreat. The patients usually have a desire to have such deformity corrected though it is not severe. In recent years we have corrected the over-conves antihelix using postauricular flap and cartilage with satisfactory results. PMID- 9206135 TI - [Basic research and clinical applications of the human hair suture]. AB - An atraumatic suture was manufactured from human hair, which has been tested and used in 817 clinical cases with satisfactory results. The authors found that the suture has the following characteristics: smooth, thin, strong and elastic, structurally steady, histocompatible, atoxic, aseptic, atraumatic, with endurance of more than one thousand times of bending. It does not swell in water or blood nor it causes reaction after being buried in tissue for 150 days. It definitely has an advantage over the silk and nylon. The rich resources of hair favor its wide applications. PMID- 9206136 TI - [The apron mucosal flap of the lower lip for the repair of the secondary deformities of the cleft lip]. AB - Twelve cases of secondary deformities of the cleft lip were repaired with the apron mucosal flap of the lower lip. The operation method is described. The advantages and disadvantages are discussed. Good results were obtained in patients with mild or medium secondary deformities. PMID- 9206137 TI - [Clinical significance of ophthalmo-electromyography in the determination of the function of the superior levator muscle]. AB - Preoperative electromyographic examinations of superior levator muscle were done in 36 ptotic patients using the Neurematic 2000 electromyograph. Shortening of the levator palpebrae superior was done through a combined internal and external route. In 12 patients of mild ptosis the spike voltage was all above 30 mv during contraction. When the spike voltage was above 100 mv, better operative results were obtained. In 24 patients whose ptosis was moderate or severe, the graphs of contraction generally showed simple phases with low amplitude. In 9 patients whose spike voltage was below 30 mv, their palpebral fissure did not reach the anticipated width after the operation. On the contrary, in 15 patients whose spike voltage was above 30 mv the operative results were satisfactory. It is concluded that in patients whose ptosis is moderate or severe, the operative procedure should be based on electromyographic examinations. If the spike voltage of the superior levator muscls is higher than 30 mv on contraction, simple shortening of the levator will be successful. If the spike voltage is lower than 30 mv and the graph presents simple phase or electro-tranquilization, then suspension of the frontalis muscle is preferable. PMID- 9206138 TI - [Early postoperative hypoxemia in infants, children and adults undergoing elective plastic surgery]. AB - Seven hundred ASA-class-1 patients undergoing elective plastic surgery were selected to study the effect of age on early postoperative hypoxemia with a pulse eximeter for continuous monitoring of arterial oxygen saturation (SpO2) in the postanesthesia recovery room. The patients were divided into four groups: group 1, 72 infants aged less than one year; group 2, 120 children aged from one to three years; group 3, 364 children aged over three years; and group 4, 144 adults aged from 18 to 58 years. The results showed that the younger the patients, the lower the SpO3 and the higher the incidence of hypoxemia in the early postoperative period. The incidences of early postoperative hypoxemia in the postanesthesia recovery room were 44.4% in group 1, 31.7% in group 2, 17.3% in group 3 and 8.3% in group 4. Early postoperative hypoxemia occurred most commonly within 40 min. in infants and within 15 min. in children aged over one year and adults. PMID- 9206139 TI - [Ventilation effects of added jet air flows in different directions during expiratory phase on dogs with steam inhalation injury]. AB - Nine dogs injured by inhaling steam were subjected to high frequency jet ventilation. Jet airflows of different directions were added during the expiratory phase, and their effects on respiration, circulation and improvment in ventilation efficiency of dogs with inhalation injury were compared, and the assessment of the mechanism of high frequency two-way jet ventilation in the trachea was made. According to the direction of added airflow during expiratory phase, the animal models were classified into three groups: Model A, the control group, without any added airflow; Model B, airflow opposite to the direction of ventilation was added; Model C, airflow of the same direction was added. The results showed: 1. Model B, with its CO2 elimination (Vco2) increased significantly (P < 0.01) and PaCO2 decreased significantly (P < 0.01), was preferable to Model A. This might be due to the reduction of dead space. 2. Model C, although there was slightly reduced anatomical and physiological dead space as compared with model A, showed worse ventilation effects. The mechanism was probably related to increase in PEEP (from 0.20 +/- 0.10 to 0.59 +/- 0.08 kPa) and FRC (from 433 +/- 59 to 504 +/- 7.0 ml). It is suggested that high frequency two-way jet ventilation is preferable for the treatment of inhalation injury with type 1 respiratory failure rather than high frequency ventilation with PEEP. PMID- 9206140 TI - [Clinical significance of determinations of blood catecholamine, glucose and isulin in burn patients]. AB - Blood catecholamine (CA), glucose and insulin were determined in 30 severe burn patients, of whom 22 were males, and 8 females. Mean burn area was 58.6 percent TBSA. These patients were in hypovolemic shock. RESULTS: CA release was persistent in severe burn patients showing two peaks in shock period and infection period (P < 0.01). The quantity of epinephrine (E) was normal, while norepinephrine (NE) was presistently high (P < 0.01), and it was over two fold of the normal value. The value seemed to reflect the severity of the injury. When CA did not show a rise in the shock period, or lowered abruptly in the sepsis period, the patients were in high risk. Increasing CA produced increase in gluconeogenesis with insufficiency in SI secretion. PMID- 9206141 TI - [Advances in the research on implanted tissue of rhinoplasty materials and its tissue substitutes]. PMID- 9206142 TI - [Enterogenous infections]. PMID- 9206143 TI - [Intestinal lymphatic circulation is one of the important portals for microbial translocation after thermal injury]. AB - The goal of this study was to determine whether the bacteria and endotoxin from GI tract could pass through the intestinal lymph circulation to the systemic circulation after severe thermal injury. Lymphatic fistula of intestine was created in 46 Wistar rats the rats were then divided randomly in scald and control groups. In scald group, animals were sustained with 30% TBSA full thickness scald. The items studied were, the dynamic changes in intestinal lymph endotoxin level, the qualitative and quantitative bacterial culture, and pathological alterations in ileal mucosa within 24 hours postburn. Results showed that the levels of intestinal lymph endotoxin and the positive culture rate and counting of bacteria were evidently increased, and ileal mucosal lacteals were dilated and epithelial cells were necrotic and denudated in scald group. These suggested that the intestinal lymph circulation is one of the important portals for endotoxin and microbial translocation after severe thermal injury, and the intestinal mucosal damage is an important factor in pathogenesis. PMID- 9206144 TI - [An experimental study on injury of intestinal immuno-barrier in rat after scald]. AB - A dynamic observation on injury of intestinal immuno-barrier in scalded rat was performed to investigate the relationship between the injury of intestinal immuno barrier and bacterial translocation. After 40% TBSA third degree scald, a decrease of IgA in intestinal content and the number of CD3+ and CD4+ T lymphocyte, and reduction of IgA coat rate of intestinal bacteria were observed. At the same time, an increase in the incidence of bacterial translocation was detected. The results indicated that intestinal immuno-barrier was damaged and its protective function was weakened after an extensive thermal injury, and it suggested that the injury of intestinal immuno-barrier might play an important role on the development of postburn bacterial translocation and postburn sepsis. PMID- 9206145 TI - [The changes in mucus components of intestine after burn and their relationship with bacterial translocation]. AB - A dynamic observation at 0.5 hr, 1 hr, 6 hr, 12 hr and 24 hr after 25% third degree burn in 363 rats: 1) The lesions of mucous membrane of ileum. 2) Assessment of the depth of mucus layer and the contents of protein, hexose and sialic acid. 3) Estimation of the content of mucus IgA, serum IgA and the number of plasma cell producing IgA in mucous membrane. 4) Bacterial culture of mesentery lymph node. The following results were achieved: 1) Serious lesions were discovered in ileal mucous membrane. 2) Mucus IgA content was reduced markedly and the reduction was related to bacterial translocation. PMID- 9206146 TI - [Observation and analysis of postburn changes in substance P (SP) and SP peptidergic nerve fibers in the intestine of rat]. AB - Substance P (SP) was measured by radioimmunoassay (RIA) and SP peptidergic nerve fibers were identified by Immunohistochemistry (IHC) in the intestinal wall of rats (TBSA 30% and with full-thickness burn) during 72 hours after burn. The results were as follows: (1) SP increased significantly at 1 hr and 4 hr postburn, then decreased markedly 8 hr later, and the low was maintained up to 72 hr. (2) The significant changes in SP peptidergic nerve fibers in the villi after burn were found with immunohistochemical studies including the morphology, distributive densities and SP contents of the SP peptidergic nerve fibers. The results from image analysis showed that the distributive densities of the nerve fibers elevated distinctly in 1 hr group, then dropped at 8 hr and 12 hr and finally elevated again. The results indicated that, the SP release might occur leading to SP consumption in the intestinal wall of rats after burn. It is possible that SP contributes to the postburn intestinal lesion in rats. The SP peptidergic nerve fibers in villus have a direct effect on the damage of intestinal mucosal barrier. PMID- 9206147 TI - [Enhancement of gut immune function by early enteral feeding enriched with L glutamine in severe burned miniswines]. AB - In order to investigate the effect of L-glutamine on gut immune function, 14 miniswines with 30% TBSA full thickness burns were randomly and equally divided into NON-GLN group and GLN group. GLN group animals were supplied with L glutamine 0.64 g/day, and NON-GLN group received equal amount of non-glutamine amino acids. The S-IgA concentration of jejunal and IgA concentration of arterial blood were determined on PBD (post burn day) 1, 4, 7, 10. S-IgA concentration of ileal contents was measured on PBD10. The results showed that the concentrations of S-IgA in the jejunal and ileal contents as well as IgA in arterial blood decreased significantly after burns in NON-GLN group. L-glutamine supplement increased the excretion of S-IgA of intestinal mucosa in the GLN group. This result suggests that oral feeding of L-glutamine improves the intestinal S-IgA excretion after burns efficiently, and it plays an important role in the prevention of endotoxin and bacterial translocation after burns. PMID- 9206149 TI - [Resection of protruding mandibular angle through combined external and intraoral approach]. AB - Mandibular angle protrusion may cause excessive width of the mandible and a disproportion of the face. A new method was introduced to resect the protruding mandibular angle through a combined external and intraoral approach. Good results were received in 5 cases. This operation avoided external scar and the danger of damage to the marginal nerve. PMID- 9206148 TI - [The influence of pharmacological manipulation on postsurgical labial scar contraction and craniofacial growth in rabbits]. AB - This study was designed to test the influence of pharmacological manipulation on scar contraction and craniofacial growth in rabbits after repairing surgically created lip defects by wide undermining of soft tissue. Two pharmacological agents, i.e. papaverine hydrochloride and hyaluronidase, were injected subcutaneously and submucosally in the wound. The preliminary results of this study indicated that the treatment with either papaverine or hyaluronidase for four weeks following lip repair with soft tissue undermining significantly reduced lip pressure and decreased tension of labial wound. Analysis of the data from the lateral head cephalography revealed that the rabbits treated with such pharmacological agents exhibited significantly more anteroposterior growth than the rabbits not treated with such agents. PMID- 9206151 TI - [Cleft palate repair with a combined method of mucosal flap pushback of the hard palate]. AB - From January of 1992, we applied a combined method to repair cleft palate in 20 patients and received satisfactory results. The method is characterized by pushing back the mucosal flap of the hard palate, a Z-plasty on the nasal mucosa, repositioning the levator muscle to lengthen the palate, circumferential pharyng oplasty using denervated extensor hallucis brevis muscle, without making relaxing incisions and elevating the mucoperiosteal flap, avoiding interference to the greater and lesser palatine vessels and nerves, without relaxing palatal aponeurosis. The advantages of this method are preserving the normal anatomy and function of the palate and nasopharyngeal cavity, improving the function of velopharyngeal closure and minimizing secondary deformities. PMID- 9206150 TI - [Correction of alveolar cleft by bone graft in the patients with cleft lip and palate]. AB - Treatment of alveolar cleft using bone graft has been developed to the stage of team approach. We summarized the clinical material of 80 cases treated by bone grafting. This study revealed that bone grafting for cleft alveolar repair was an effective procedure valuable to be popularized. Properly forming the bed of bone grafting and perfectly closing the recipient site is the key to a successful operation. Patients should accept orthodontic treatment before and after the operation. We discuss the indications, technique, result evaluation and management before and after the operation. PMID- 9206152 TI - [Middle ear function in children with cleft palate]. AB - Middle ear problems in patients with cleft palate have for a long time received insufficient attention. The present study was designed to evaluate middle ear function in 56 children with cleft palate and in a control group of 50 children without cleft palate. The age ranging between 5 and 14 years was similar in both groups. All children underwent routine otoscopic examinations of ear, nose and throat, and were also examined using the Madsen acoustic impedance audiometer. Four major parameters (static compliance, middle ear pressure, acoustic stapedius reflex and Eustachian tube function) were analyzed. It was found that the negative middle ear pressures were higher and the static compliance, which reflected the elasticity of the conducting mechanism of the middle ear, was poorer in children with cleft palate than in the controls. The proportion of children with cleft palate who showed the presence of the acoustic stapedius reflex was also lower than in the control children. Children with cleft palate have a limited ability to actively open the Eustachian tube by swallowing, as evidenced by a poor ability to equilibrate applied positive or negative middle ear pressure. A multidisciplinary approach should be adopted for the complete overall rehabilitation of children with cleft palate. Plastic surgeons to whom most of the treatment is entrusted should be concerned not only with achieving palatal function and acceptable speech, but also with the various other problems associated with cleft palate, especially those affecting the middle ear. PMID- 9206153 TI - [Applications of the island trapezius myocutaneous flap in oral maxillofacial surgery]. AB - The experience in immediate reconstruction of oral maxillofacial defects of 23 cases by island trapezius myocutaneous flaps from April 1985 to July 1993 is presented. The smallest flap was 6 x 4 cm and the largest 12 x 8 cm, with all the cases successful. The selection of the vascular pedicle of the myocutaneous flaps, the technique of the operation, the advantages and the disadvantages of the method are discussed. PMID- 9206154 TI - [Congenital anotia with hemifacial atrophy (a report of 12 cases)]. AB - Twelve cases of congenital anotia with hemifacial atrophy were examined and analyzed with electromyogram and roentgenography. The results revealed that the development of the deformed side of the face was 30% less than the other side. Because both the soft tissue and the skeleton of the deformed side are undeveloped, they should be dealt with simultaneously. PMID- 9206155 TI - [Effects of electret and Ligusticum wallichii (chuangxiong) on the functional recovery of muscle grafts]. AB - The aim of this study was to evaluate the effects of electret and Ligusticum wallichii (Chuangxiong, a traditional Chinese herb medicine) on the functional recovery of muscle grafts. Twenty-eight adult dogs were divided into 4 groups with each consisting of 7 animals. After orthotopical transplantation of bilateral rectus femoris muscle with neurovascular anastomosis, group A received electret local implant, group B received Chuangxiong injection, group C received both electret local implant and Chuangxiong injection and group D was used as control. The results showed that the functional recovery of the transplanted rectus femoris muscle in group A and B was better than group D while group C was the best. At 22 postoperative weeks, the recovery rate of maximal tetanic tension was 64.94 +/- 3.82% in group c: 57.68 +/- 1.67% in group A, 53.64 +/- 3.82% in group B and 47.99 +/- 2.21% in group D (P < 0.05). The structural recovery of the transplanted muscle in group C was closest to the normal. PMID- 9206156 TI - [The turn-over flap of the frontalis muscle used for eye-socket depression with contraction of the conjunctival capsule]. AB - We were used to repair the eye-socket depression and contraction after eyeball loss with fat, dermis or rib cartilage implantation. This "stuffing method" has some disadvantages, including absorption and exposure of the implant. In recent years the authors have used a turn-over flap of the frontalis muscle to treat eye socket depression with contraction of the conjunctival capsule. Satisfactory results have been found at postoperative follow-up. PMID- 9206157 TI - [A preliminary study on correcting the high fold of the upper eyelid]. AB - Since 1989 eighteen patients (23 eyes) with high folds of the upper eyelid have been treated by sutruing the orbital septum, pretarsal portion and the incisions together. It created a barrier between the high upper palpebral fold and the levator palpebrae superioris tendon. After the operation, the high fold disappeared and double-fold eyelid was better formed. The mechanism of the technique is discussed. PMID- 9206158 TI - [The repair of heel defect]. AB - A total of 12 lateral leg complex flaps with gastrocnemius muscle, lateral cutaneous nerve of calf, cortex of fibula have been applied successfully to repair the heel defects. The walking and weight carrying function of feet were recovered. The author think that repair of heel defect might base on the anatomical, physiological function of heel to choose the appropriate complex flap. PMID- 9206159 TI - [Mycethemia: an analysis of 56 cases]. AB - Mycethemia was diagnosed in 56 burn patients between 1958 and 1992. The overall incidence was 0.39% and the mortality rate was 39.29%. Candida albicans, constituting for 75% of cases, was most common encountered. The incidence was 0.26% and the mortality 72.73% in years before 1985, while that in the years after 1985 were 0.58% and 17.65%, respectively. The differences are of significance (P < 0.01) predisposing. The clinical characteristics were summed up, and the causes, clinical diagnosis and treatment were studied and discussed. PMID- 9206160 TI - [A study of immune suppression activity of serum from burned rabbits]. AB - A burned rabbit's model (20% TBSA) was used in this study. The sera taken from the burned rabbits without bacteremia and sepsis showed immune suppressive effects, especially the sera obtained 4 days after injury. After being separated by ultrafiltration, the burned sera were divided into three groups. The ultrafiltrating substances with molecular weight below 10 Kd or between 10 to 30 Kd were found to reduce the production of IL-2 and the lymphocyte blastogensis stimulated by Con A. The substances with molecular weight greater than 30 Kd also showed profound immune suppressive action on lymphocyte. No obviously abnormal protein was found in the burned serum analysed with SDS electrophoresis. It is concluded that the immune suppressive substances in the burned serum were characterised by complexity in components and a wide range of molecular weight. PMID- 9206161 TI - [Neutrophils and ischemia reperfusion injury]. PMID- 9206162 TI - [Current situation, controversy and future of gynecological laparoscopic surgery]. PMID- 9206163 TI - [Laparoscopic diagnosis of tubal infertility and fallopian tube lesions]. AB - OBJECTIVE: To analyse the diagnostic value of laparoscopy in patients with tubal infertility. METHODS: The morphological appearance of fallopian tubes and surrounding tissues of 1120 cases with proven tubal infertility were observed under laparoscopy. Hydrotubation was performed at the same time in each case. RESULTS: Tubal infertility diagnosed by laparoscopy accounted for 32.8% of infertile patients. Among them, pelvic tuberculosis occupied 63.6%, while nonspecific inflammatory disease (NSID) 36.4%. 44.8% of the tuberculosis and 62.2% of NSID group had negative findings during pelvic examination. Four types of tuberculosis lesions were demonstrated: miliary ascites (9.4%), adherent mass (35.8%), adhesion and calcification (43.1%), nodular sclerosis (11.7%). In the NSID group, simple tubal obstruction accounted for 29.9%, the remaining were mild adhesion or hydrosalphinx. Complete tubal occlusion occupied 81.2% in tuberculosis group and 70.7% in NSID cases. In the tuberculosis group the positive rate of pelvic lesion biopsy and endometrial biopsy was only 59.1% and 20.5% respectively. CONCLUSION: Laparoscopy examination is a valuable procedure for the etiological diagnosis of tubal infertility. PMID- 9206164 TI - [Analysis of complications following laparoscopic surgery]. AB - OBJECTIVE: To analysis the causes, management and prevention of complications following gynecological laparoscopic surgery. METHOD: We analysed retrospectively the clinical data of 22 cases with complications following laparoscopic surgery performed from Sep 1992 to May 1994. RESULTS: The overall incidence of operative complications was 6.29% (22/350). Among them, 9 cases (including pneumoderma 6 and pneumo-omentum 3) were related to inappropriate CO2 insufflation. The other 10 cases were associated with bleeding, i.e. vessel injury 4 cases, hypodermal ecohymosis 5, and bleeding from the sutured site 1. The remaining 3 were omental or intestinal hernia of the incisional sites. One out of the 22 cases were turned to laparotomy. The outcomes of this group were all good. CONCLUSIONS: The occurrence of these complications was related to the doctors experiences and skills. Therefore, well-training and practice should be emphasized in order to avoid these complications. PMID- 9206165 TI - [Diagnosis of uterine diseases by combined hysteroscopy and ultrasonography]. AB - OBJECTIVE: To investigate the diagnostic value of combined hysteroscopy with ultrasonography. METHODS: 477 patients were diagnosed by hysteroscopy, ultrasonography, or hysteroscopy combined with ultrasound. The results of the 3 groups were compared with the findings after operations. RESULTS: This study indicated to diagnose intrauterine lesions, intramural diseases or pelvic disorders, hysteroscopy with ultrasonography was significantly better than either methods alone. The accuracy rate, false positive rate as well as false negative rate were 98.32%, 1.26% and 0.04% respectively. CONCLUSION: Hysteroscopy is used for diagnosis and treatment of intrauterine lesions, and ultrasonography is difficult to show actual location of lesions within uterine cavity. The present study has shown that hysteroscopy combined with ultrasound could increase the effectiveness and accuracy of uterine diseases' diagnosis. PMID- 9206166 TI - [Investigation of urinary fibrinopeptide A level in pregnancy induced hypertension and its clinical significance]. AB - OBJECTIVE: To investigate the changes of urinary fibrinopeptide A (FPA) level in pregnancy induced hypertension (PIH) and its clinical significance. METHODS: High performance liquid chromatography (HPLC) was employed in determination. Urinary FPA was measured in 96 cases of normal pregnancy and 49 cases with PIH. RESULTS: The urinary FPA levels in mild and moderate PIH groups were significantly higher than that in normal pregnancy (71.65 +/- 18.53 micrograms/L vs 40.17 +/- 20.26 micrograms/L, P < 0.01). The levels of urinary FPA in patients with preeclampsia increased significantly (146.65 +/- 32.53 micrograms/L) when compared with the mild and moderate PIH groups (P < 0.001). The levels of urinary FPA in patients with eclampsia (422.93 +/- 81.46 micrograms/L) were much higher than that in group of preeclampsia (P < 0.001). CONCLUSIONS: It showed that the assay of urinary FPA might be helpful for earlier diagnosis, classification, treatment and judgement of prognosis in PIH. PMID- 9206168 TI - [Pulsatility indexes of fetal middle cerebral artery and umbilical artery for predicting intrauterine fetal growth retardation]. AB - OBJECTIVE: To evaluate the role of fetal cerebral and umbilical blood flow for predicting intrauterine fetal growth retardation (IUGR). METHODS: Doppler ultrasonography was used to study pulsatility indexes (PI) of the fetal middle cerebral artery (MCA) and umbilical artery (UA) in 84 normal late pregnancies and 31 IUGR cases. RESULTS: MCA PI were significantly lower in IUGR fetuses than that of normal fetuses (P < 0.01). UA PI and UA PI/MCA PI ratio were higher in IUGR group than that of normal group (P < 0.01). The sensitivities of MCA PI, UA PI and UA PI/MCA PI ratio for predicting IUGR were 80.64%, 70.96% and 87.09% respectively at the cut off level with 2 standard deviation (SD). The specificities were 94.05%, 88.90% and 97.61% respectively. CONCLUSIONS: Doppler ultrasonography for predicting IUGR was a practical and sensitive method. PMID- 9206167 TI - [The relation between indirect fetal electrocardiogram and blood gas analysis of umbilical cord artery during labor]. AB - OBJECTIVE: To observe the relation between indirect fetal electrocardiogram (FECG) and blood gas analysis of umbilical cord artery blood during labor and discuss the possibility of using FECG for labor monitoring. METHODS: Indirect FECG was used for fetal monitoring in 80 cases during the second stage of labor and cord umbilical artery blood was taken immediately after delivery for blood gas analysis. Cases were retrospectively divided into normal and abnormal groups according to the results of FECG. RESULTS: The success rate of FECG test was 91.95%. Significant differences were noted in mean values of pH, PCO2, PO2, actual base excess (ABE) and standard base excess (SBE) of umbilical artery between the 2 groups, so were the percentages of cases with pH < 7.20, PCO2 > 8.00 kPa, PO2 < 2.10 kPa. CONCLUSIONS: Indirect FECG can be used for fetal monitoring during labor. FECG is obviously correlated with acid-base equilibrium and blood gas concentration of umbilical cord artery blood, it is a sensitive index of fetal and neonatal hypoxia. PMID- 9206170 TI - [Immunotherapy of primary habitual abortion]. AB - OBJECTIVE: To investigate whether different number of times of immunization and different immunogens have any effect on the outcome of pregnancy in patients with history of primary habitual abortion. METHODS: 38 cases of pregnant women with history of primary habitual abortion immunized with lymphocytes were analysed retrospectively. RESULTS: The results showed that the successful pregnancy rate was 87.5% (14/16) in the women immunized two times and was 86.4% (19/22) in the women immunized four times. There was no significant difference between the two groups. The successful pregnancy rate was 84.2% (16/19) in women immunized with their husbands' lymphocytes and 89.5% (17/19) in women immunized with donors' lymphocytes. There was also no significant difference between the two groups. CONCLUSIONS: In order to prevent hematogenous infection, active immunization could be reduced from four to two times and the immunogens may be selected from the husband or from healthy donor. PMID- 9206169 TI - [Rapid prenatal diagnosis of 60 cases with beta-thalassemia by reverse dot blot analysis]. AB - OBJECTIVE: To evaluate the rapid prenatal diagnosis method for beta-thalassemia by reverse dot blot (RDB) analysis. METHODS: Using RDB analysis which can detect 18 types of beta-thalassemia mutation, 60 fetuses (with 1 twin) suspected of beta thalassemia from 59 couples in Southern China were investigated. RESULTS: 15 fetus had normal genotype, 34 were carriers of heterozygous with one of parental affected gene; 10 had both parental affected genes-heterozygous or homozygous; and 1 was not able to identified. Three rare types of beta-thalassemia mutations (CD43, CD14-15, CD27-28) were detected. CONCLUSIONS: The RDB assay is able to complete the screening of beta-thalassemia mutation by DNA hybridization in only one single working day. It is simple, accurate and convenient to operate without radio-isotopes. It can be used as a routine technique in clinical laboratory for the detection of carriers and prenatal diagnosis of beta-thalassemia. PMID- 9206171 TI - [The study of immunotherapy of ovarian carcinoma with anti-idiotypic antibody]. AB - OBJECTIVES: To study the role of anti-idiotypic antibody in immunotherapy of ovarian carcinoma, and to investigate the effect of anti-idiotypic antibody (Ab2) on ovarian carcinoma in vivo. METHODS: Six to eight week-old female BCF1 mice were immunized three times at 28 days intervals with 50-100 micrograms of anti idiotypic antibody Ab2 of ovarian carcinoma. The initial immunization was given with complete Freund's adjuvant (CFA), followed by incomplete Freund's adjuvant (IFA). The control mice were immunized with normal mouse IgG. The ovarian carcinoma cells (SKOV3) passaged from nude mice were transplanted to the subrenal capsules of immunized BCF1 mice after the last injection. Then the mice were bred separately on the 2nd, 4th, 6th, 8th and 10th days after the transplantation operations. Specific anti-Ab2(Ab3) in sera was detected by using inhibition assay of enzyme-linked immunospecific assay. Tumor infilitration lymphocytes (TIL) and tumor cells from the tumor tissue of BCF1 were observed microscopically. RESULTS: The sera levels of Ab3 in Ab2 immunized mice were higher than those of the controls. TIL in the former were found to reach their highest peak on the 6th day, where as the highest peak for the controls was on the 8-10th days, tumor cell viability rate in the former decreased rapidly after 6 days, but in the controls decreased only gradually. CONCLUSIONS: Ab2 could induce specific immuno rejection to ovarian carcinoma in vivo. It is predicted that anti-idiotypic antibody Ab2 might be used in immunotherapy of ovarian cancer patients and also as a preventive for post treatment clinical recurrence of ovarian carcinoma. PMID- 9206172 TI - [Influence factors in etiology of epithelial ovarian cancer]. AB - OBJECTIVE: To investigate the role of genetic and psychosomatic factors in the development of epithelial ovarian cancer. METHODS: A hospital-based case-control study was conducted in 9 hospitals of Shandong province. 127 cases of epithelial ovarian cancer and 254 cases of non-malignant ovarian tumors were admitted for treatment during the same period of time. Data for single predictors and multivariants collected from hospital records were subjected to non-condition multivariate logistic analysis, in order to sort the protective factors and the high risk factors in the development of epithelial ovarian cancer. RESULTS: Adverse events in life, blood type A, mental depression or anger, family history of ovarian cancer and irregularity of bowel movements are the high risk factors. An open and cheerful disposition or optimism, hysterectomy with preservation of one or both ovaries and intrauterine device were shown to be the protective factors for the ovaries. CONCLUSION: Genetic and psychosomatic factors play an important role in the course of development of epithelial ovarian cancer. PMID- 9206173 TI - [Laparoscopic surgery in benign ovarian cysts]. PMID- 9206174 TI - [Evaluation of the quality of life in patients with gynecologic cancer and its intervention]. PMID- 9206175 TI - [Standardization in the treatment of gynecologic malignancies should be emphasized]. PMID- 9206176 TI - [Recent trends in the diagnosis and treatment of endometriosis, postmenopause and assisted reproductive technique]. PMID- 9206177 TI - [Summary of the Sixth National Conference on Obstetrics and Gynecology]. PMID- 9206178 TI - [Fetal erythroblasts from maternal blood for prenatal genetic diagnosis]. AB - OBJECTIVE: To study fetal erythroblasts (FE) from maternal peripheral blood for the diagnosis of fetal aneuploidies. METHODS: FE expressing the glycophorin A(GPA) were isolated from 13 pregnant women with male fetus (8-14 w) by fluorescence-activated cell sorting(FACS), FE were identified by oligonucleotide primed in situ labelling (PRINS) with Y centromeric satellite DNA primer. The concentration of pregnancy-associated plasma protein A (PAPP-A) was measured by enzyme-labelled immunosorbent assay (ELISA) in serum samples of 41 normal pregnant women (8-14 w). In 5 pregnant women suspicious of fetal Down's syndrome (10-13 w) the serum and FE were examined by PAPP-A, GPA/FACS and PRINS with 21 chromosome centrometric primer. RESULTS: Detection of flow sorted FE from 13 pregnant women by Y primer showed 14.5% of GPA positive signal. There was no difference in serum level of PAPP-A between 5 pregnant women and 41 normal controls, and all GPA positive cell nuclei of the 5 cases displayed two signals with 21 chromosome. CONCLUSION: Measurement of fetal erythroblasts from maternal blood for the diagnosis of genetic fetal aneuploidies is a promising non invasive, rapid and reliable technique. PMID- 9206179 TI - [Metabolism of angiotensin I in placenta tissue]. AB - OBJECTIVE: To explore whether placenta tissue could produce angiotensin I and the metabolism pathway of angiotensin I in placenta. METHODS: Placenta tissues from 6 term pregnant women without any complication were cultured in vitro for 1 to 6 days, the angiotensin I concentrations in the medium were measured by radioimmunoassay in the first, second, fourth and sixth day. 5 to 500 mumol/l, captopril were added to the culture medium and the concentrations of angiotensin I were measured at the same times to observe the effect of angiotensin converting enzyme on the production of angiotensin I. RESULTS: The concentration of angiotensin I in the placenta culture medium increased with the culture time. There were no significant differences in the concentrations, when captopril was added. CONCLUSIONS: The placenta tissue can produce angiotensin I and its production does not depend on the angiotensin converting enzyme. PMID- 9206180 TI - [Hypergonadotropic secondary amenorrhea: clinical analysis of 126 cases]. AB - OBJECTIVE: To study the etiology and early diagnosis of hypergonadotropic amenorrhea and to explore the appropriate treatment for preserving their reproductive function. METHODS: 126 cases of secondary amenorrhea with serum follicular stimulating hormone (FSH) levels > or = 40 IU/I, were analysed. Their clinical manifestations, karyotypes, ovarian morphology and histology, reproductive hormone assays, and responses to estrogen therapy and ovulation induction were studied. RESULTS: 6 cases presented with histories of ovarian surgery, radiotherapy or chemotherapy. Among the other 120 cases, 18 manifested amenorrhea before or at 25 years of age, 102 developed amenorrhea after age 25. In the former group, 16 (88.9%) showed unilateral or bilateral gonadal dysgenesis, and the other 2(11.1%) were defined as resistant ovaries. Abnormalities of sex chromosome karyotype occurred in 44.4% (8/ 18). In the latter group, 68 underwent laparotomy or laparoscopy examination. Morphological and histological examinations of both ovaries showed atrophic ovaries in all cases accompanied by 30.9% (21/ 68) unilateral gonadal dysgenesis; sex chromosomal abnormality was found in only one with no sexual immaturation. The efficacy of estrogen treatment was significantly better among cases with amenorrhea less than 1 year as compared with those longer than 1 year. Clomiphene challenge test given to 8 cases during their irregular menstrual stages produced an elevation of FSH levels to > 20 IU/I. without any response of estradiol secretion. CONCLUSIONS: The earlier estrogen therapy is initiated, the greater possibility of pregnancy will be achieved in cases suffering from hypergonadotropic amenorrhea. The clomiphene challenge test may provide evidence of waning ovarian function for early diagnosis. PMID- 9206181 TI - [Fertilization in vitro and embryo transfer by combined long-acting gonadotropin releasing hormone agonist and gonadotropin for superovulation in patients with refractory polycystic ovary syndrome]. AB - OBJECTIVE: To investigate the efficacy of the treatment for polycystic ovary syndrome (PCOS) patients by combined long acting gonadotropin-releasing hormone agonist (GnRHa) and gonadotropin. METHODS: Nineteen women with PCOS who had failed to conceive on treatment with conventional fertilization in vitro-embryo transfer (IVF-ET) protocol underwent 26 cycles of IVF-ET by combined long-acting and gonadotropin stimulation. A self-control group consisted of 19 PCOS women who had received 19 treatment cycles with conventional IVF-ET stimulation protocol more than 6 months previously. RESULTS: A comparison of these two groups showed that treatment with combined long-acting GnRHa and gonadotropin improved the in vitro fertility rate (76.2%) and pregnancy rate (38.5%). CONCLUSION: Patients with refractory PCOS should be referred for IVF-ET, using long-acting GnRHa before gonadotropin as the stimulation protocol. PMID- 9206182 TI - [Clinicopathological evaluation on ovarian serous borderline tumours and their implants]. AB - OBJECTIVE: To investigate the clinicopathologic features and the expressions of proliferation cell nuclear antigen (PCNA) in ovarian serous borderline tumours (OSBT). METHODS: The primary ovarian tumours, implants and recurrent specimens of 48 cases of OSBT were studied. Expression of PCNA was determined by avidin-biotin peroxidase complex (ABC) method. Clinical data for all patients were abstracted from hospital records. RESULTS: While the primary ovarian tumours have the characteristic histological pictures of borderline tumour, three different types of histologic appearance including benign, noninvasive and invasive were shown by their peritoneal implants. All benign and most noninvasive implants were PCNA negative or weakly positive, and the 25 patients were free from tumour 5-28 years after their operations. Some of the noninvasive implants (9/22) were recurrent, and a few (3/22) underwent malignant transformation 10-23 years after surgery. The only one invasive implant was PCNA strongly positive and the patient died 3.5 years after surgery. CONCLUSION: The different histologic types of the peritoneal implants are of prognostic significance and are factors to be considered in determining the treatment project. The diagnostic value of PCNA immunoreactivity in implantations is confirmed. PMID- 9206183 TI - [Effects of recombinant human tumor necrosis factor on the development of ascitic tumor and expression of c-erbB2 in the nude mouse models of ovarian cancer]. AB - OBJECTIVE: To investigate the effects of recombinant human tumor necrosis factor (rhTNF) on the growth of ascitic tumor and expression of c-erbB2 in the nude mouse models of ovarian cancer. METHODS: By intraabdominal injection, rhTNF was delivered to the peritoneal cavity of nude mouse with ovarian cancer as well as control group, after 6 days treatment, tumor cell counting and expression of c erbB2 were detected by indirect fluorescence flow cytometry. RESULTS: There is a significant difference in tumor cell counting between the study group and control group (P < 0.01). Expression of c-erbB2 in study group was lower than that in control group. CONCLUSION: rhTNF was demonstrated to have effects on both inhibition of ascitis tumor development and down regulation of c-erbB2 expression in nude mouse with ovarian cancer. PMID- 9206184 TI - [Early prediction of fetal growth retardation by umbilical and uterine arterial flow velocity systolic to diastolic ratio]. AB - OBJECTIVE: To evaluate the umbilical and uterine arterial Doppler flow velocity waveform systolic to diastolic (S/D) ratios performed at 24-30 weeks gestation for predicting fetal growth retardation (IUGR). METHODS: A prospective double blind study was conducted in 118 cases of high risk singleton pregnant women. The umbilical and uterine arterial S/D ratios were measured at 24-30 gestational weeks and the pregnancy outcomes were followed up. RESULTS: The prevalence of IUGR in our study population was 16.9%. At 24-30 weeks gestation, the S/D ratio of both umbilical artery and uterine artery in IUGR pregnant women were significantly higher than those in normal pregnant women, while the fetal biometric measurements were normal in all the 118 cases. The sensitivity, specificity and positive predictive value of umbilical arterial S/D ratio to predict IUGR were 80.0%, 83.7% and 50.0% with a Kappa index of 0.51 at 24-30 weeks gestation. With lower sensitivity, specificity, positive predictive value and Kappa index (40.0%, 84.5%, 34.8% and 0.23 respectively), the uterine arterial S/D ratio had less predictive value. CONCLUSIONS: The umbilical arterial Doppler flow velocity waveform S/D ratio may be an earlier predictor for screening of IUGR at 24-30 weeks gestation in high risk pregnant women with normal fetal biometric measurements. PMID- 9206185 TI - [Determination of platelet activating factor and platelet associated IgG in pregnancy-induced hypertension]. AB - OBJECTIVE: To determine the levels of platelet activating factor (PAF) and platelet associated IgG (PAIgG) in pregnancy-induced hypertension (PIH). METHODS: Twenty-eight cases of normal pregnant women and 44 cases of PIH (moderate, n = 23; severe, n = 21) were enrolled in this study. The plasma level of PAF was measured using platelet aggregation bioassay, and determination of PAIgG was performed by ABC-ELISA (enzyme-labeled immunosorbent assay) method. RESULTS: The level of plasma PAF in PIH patients (5.02 +/- 1.93 micrograms/L) was significantly higher than that in the controls (1.16 +/- 0.22 micrograms/L, P < 0.01). There was no difference between the levels of PAF in moderate PIH and severe ones. The level of PAIgG in PIH (6.3 +/- 4.9 micrograms/10(2)) was significantly higher than that in the control (3.8 + 1.8 micrograms/10(2), P < 0.05). CONCLUSIONS: PAF may be involved and play an important role in the pathophysiological changes in PIH. Autoimmunological reaction of platelets may exist in pregnant women with PIH. PMID- 9206187 TI - [Safety of 5% dextrose as distending medium during transcervical resection of endometrium]. AB - OBJECTIVE: To observe the changes of plasma electrolytes and osmolarity during and after transcervical resection of endometrium using 5% dextrose solution as distending medium. METHODS: We used 5% dextrose solution as distending medium for 500 cases of operative hysteroscopy. Plasma sodium, potassium, chloride, glucose concentrations and plasma osmolarity were measured before, at the end of 1 hour, 4 hours and the day after operation respectively in 22 cases. RESULTS: None of the 500 cases presented with transurethral resection of prostate (TURP) syndrome. No apparent change were found in plasma sodium, chloride concentrations and plasma osmolarity. Plasma potassium concentration decreased gradually but significantly 4 hours after surgery. The elevation of plasma glucose concentration was significant and was positively correlated with the absorption of 5% dextrose. All changes of plasma electrolytes and osmolarity restored in the next morning. It was 1 hour after surgery that the lowest plasma sodium, potassium concentration as well as the highest plasma glucose concentration appeared. Therefore, it is important to observe patient 1 hour after operation. CONCLUSIONS: 5% dextrose solution, used as the distending medium, is safe and inexpensive, and could be widely applied in transcervical resection of endometrium. PMID- 9206186 TI - [Effect of dihydroetorphini hydrochloride on mice parturition]. AB - OBJECTIVE: To determine the effect of dihydroetorphini hydrochloride (DHE) on parturition of mice. METHODS: 180 pregnant mice were divided equally into 6 groups. DHE was administered intramuscularly (i.m.) to mice group A, B and C at the dose levels of 1, 2 and 4 micrograms/kg respectively. Morphine hydrochloride 200 micrograms/kg (hypodermic) was given to group D and procaine hydrochloride 1000 micrograms/kg i.m. for group E, while for group F as control distilled water was given. Each preparation was given to the pregnant mice 3 times, i.e. 18 hours before the date of confinement, during labor and 2 hours postpartum. RESULTS: No effects on mice parturition were found in group A, B, E, and F. The percentage of cyanosis in newborn mice of group C and D was 13.8% and 22.8% respectively, which was significantly higher than that (7.4%) in group F. CONCLUSION: DHE is safer than morphine and procaine for analgesia during labor, and no adverse effects of DHE at dose levels of 1 microgram/kg and 2 micrograms/kg were observed on mice parturition. PMID- 9206188 TI - [Ovarian metastasis in uterine cervical cancer: analysis of 17 cases]. AB - OBJECTIVE: To investigate the incidence of ovarian metastasis in cervical cancer, especially in early FIGO stages. METHODS: Seventeen cervical cancer patients with ovarian metastasis treated at Cancer Hospital, Chinese Academy of Medical Sciences from 1958 to 1994 were retrospectively analysed. The diagnoses of all patients were confirmed by operations and pathologic examinations. RESULTS: The incidence of ovarian metastasis in cervical cancer was 0.07% in squamous cell carcinoma and 1.81% in adenocarcinoma (P < 0.05). In early stages (stage l-I), the incidence of ovarian metastasis in adenocarcinoma was higher than that in squamous cell carcinoma (2.2% VS 0.08%, P < 0.05). In patients treated by surgery, the rate of ovarian metastasis in adenocarcinoma was also higher than that in squamous cell carcinoma (7.8% VS 1.6%, P < 0.05). 10 of the 17 cases had macroscopic metastasis and 7 had microscopic metastasis. 11 patients (58.8%) were with bilateral ovarian involvement. Uterine corpus invasion and retroperitoneal lymph node involvements were high risk factors for ovarian metastasis in cervical cancer. The prognosis of cervical cancer patients with ovarian metastasis was poor, the 5-year survival rate in this study being only 17.6%. CONCLUSIONS: Patients with cervical cancer are threatened with the risk of ovarian metastasis, especially in adenocarcinoma. Conservative treatment of the cervical cancer with reservation of the ovary should be further investigated. PMID- 9206189 TI - [The study of radioimmunoimaging on the nude mice bearing human ovarian carcinoma by labelling monoclonal antibody with 99mTc]. AB - OBJECTIVE: To investigate the feasibility of radioimmunoimaging (RII) on the nude mice model by labelling anti-ovarian carcinoma monoclonal antibody (McAb, COC166 9) with 99mTc, and the possibility for clinical application. METHODS: McAb reduced by beta-mercaptoethanol was labelled directly with 99mTc. Imaging positive rate, negative rate and tumor/non-tumor (T/N) ratio were calculated. RESULTS: (1)McAb COC166-9 labelled with 99mTc reacted with ovarian carcinoma. (2) 8 nude mice models were injected with the labelled antibodies and were then imaged. In the experimental group the positive imaging rate of tumor was 100%, in the control group the negative rate was 100%. The results were the same as those of T/N ratios. (3)In two groups (10 nude mice), intraperitoneal (i.p.) or intravenous (i.v.) injection of McAb labelled with 99mTc, positive imaging rates of tumor 100%. The IV group was better than IP group in the radio-background. CONCLUSIONS: Immunoactivity of 99mTc-COC166-9 used for RII on the nude mice models was good, and the imaging effect was specific. Clinical application is promising. PMID- 9206190 TI - [Some thoughts on clinical family planning studies]. PMID- 9206191 TI - [A phase III multicentre study on medical termination of early pregnancy with two regimens of mifepristone followed by PG05]. AB - OBJECTIVE: To compare use-effectiveness and side-effects of medical termination of early pregnancy by oral administration of separated doses of mifepristone 150 mg or single dose of mifepristone 200 mg followed by PG05 1 mg vaginally in 4500 healthy pregnant women with amenorrhea < or = 49 days in a phase I multicentre study. METHODS: All subjects were recruited in 30 clinics after obtaining informed consents. RESULTS: The complete, incomplete abortion and continuous pregnancy rates were 88.6%, 8.1%, 1.7% and 92.0%, 5.8%, 0.8% in single dose and separated doses groups, respectively. The efficacy of termination of early pregnancy by mifepristone 150 mg in separated doses was higher than that in single dose of mifepristone 200 mg when both in combination with PG05 1 mg (P < 0.01). One case of allergic rash was noted after administration of mifepristone. No severe side-effects were observed except bleeding problems. 65 cases received emergency vacuum aspiration and 7 of them received blood transfusion due to heavy bleeding. In addition, 2 cases of ectopic pregnancies required blood transfusion as heavy bleeding caused by exfetation disruption. There was no significant difference in to the time to start bleeding and the days of bleeding lasted between two treatment groups. However, there was a significant positive correlation (P < 0.001) between diameters of SAC and the days of bleeding lasted. CONCLUSIONS: Logistic regression analysis suggested that subject's age, gravidity and days of amenorrhea have significant effects on abortion efficacy. 75.2% of subjects are satisfactory in self-assessment of the treatment, and 81.8% of subjects will choose medical abortion method in case they need to terminate undesired early pregnancy in the future. PMID- 9206192 TI - [Vaginal bleeding patterns and the regularity in use of Norplant]. AB - OBJECTIVES: To understand vaginal bleeding patterns and the regularity in use of Norplant, and find out the main cause of termination that Norplant users can not bear. METHODS: A total of 306 menstrual diaries of Norplant users for 5 years were analyzed. The analysis of bleeding patterns was carried out by using the reference period approach and followed the guidelings published by WHO. RESULTS: The total number of vaginal bleeding days and the number of spotting days in the first reference periods were 36.6 days and 21.5 days, respectively. They were decreased obviously in the third reference period. The number of bleeding days was not obviously changed. The percentage of irregular bleeding was predominent in all of the bleeding patterns, 43.1%-57.6% usually. The percentage of prolonged bleeding was 13.4%-31.0%, secondly. The percentage of "normal" was only 33%. The number of bleeding days and the percentage of prolonged bleeding were more in bleeding termination group than in continuation group. The percentage of irregular bleeding was lower. CONCLUSIONS: During the initial stage of use of Norplant the total number of vaginal bleeding days was increased, but decreased obviously after 6 months. This change was influenced by the number of spotting days. Irregular bleeding and prolonged bleeding were main types of the disturbance of menstrual cycles after use of Norplant, The main cause of termination was prolonged bleeding. PMID- 9206193 TI - [Clinical trial of an anti-fertility method with testosterone enanthate in normal men]. AB - OBJECTIVE: To establish the feasibility of anti-fertility by ultraphysiological dosage testosterone in male. METHODS: Ten healthy, fertile men received 200 mg testosterone enanthate weekly by intramuscular injection based on international standard protocol. RESULTS: After 3-month injection, severe oligospermia was achieved. After 6-month injection, azoospermia was achieved. The total consecutive administration lasted for 12 months. During the period of severe oligospermia and azoospermia no partner was pregnant. After stopping injection, the sperm output returned to normal (20 x 10(8)/ml) in 2-3 months up to individual's own level before administration in 6 months. CONCLUSION: Severe oligospermia and azoospermia can be induced by ultra-pathological-dosage tesosterone through inhibiting FSH, LH to cause spermatogenesis inhibition. The sperm output would recover to its previous level in 6 months after stopping injection. It is suggested that there is a strong possibility that high-dosage testosterone may act as a male contraceptive. PMID- 9206195 TI - [Construction of gene library of attenuated liver hepatitis A vaccine (H2 strain)]. AB - OBJECTIVE: To determine the complete gene library of attenuated live hepatitis A vaccine (H2 strain) from cloned cDNA by application of reverse transcription (RT) and polymerase chain reaction (PCR) technique. METHODS: Attenuated HAV H2 strain that was selected and studied in China was proven attenuated for human beings and proved to be effective in preventing hepatitis A. RESULTS: Ten overlapping cDNA clones were obtained in addition to spanned the entire genome. The restriction enzyme mapping was also determined. CONCLUSION: The gene library will benefit for the sequence of HAV cDNA and study on attenuation. PMID- 9206194 TI - [DNA typing for HLA-DR in donor-recipients of cadaveric transplantation]. AB - OBJECTIVE: To accurately allocate donor-recipients of HLA-matching and improve long-term graft survival, genotyping method for HLA-DR alleles by polymerase chain reaction with sequence-specific primers (PCR-SSP) was established and applied to renal transplantation. METHODS: Thirty primers were designed and synthesized according to the HLA-DR nucleotide sequences. Genomic DNAs were prepared by a rapid salting-out method. A rapid genotyping method of PCR-SSP was set up by PCR technique and applied to HLA-DR typing in 14 cell lines DNA and 171 individuals of donor-recipients of cadaveric transplantation. The amplification was accomplished by 34 cycles consisting of denaturation at 94 degrees C for 30 seconds, annealing at 60 degrees C for 50 seconds, and extension at 72 degrees C for 40 seconds. The specificity of matching was determined against a panel of standard DNA, analysis with restriction endonucleases and Southern hybridization. RESULTS: HLA-DR alleles of all 171 samples and 14 cell line DNAs were able to be typed by PCR-SSP. The size of specific products was consistent with the size of calculation. The overall time of genotyping was only 5 hours. No false positive or false negative typing results were discovered. The typing results were confirmed by analysis with endonucleases and hybridization. The specificity and reproducibility were 100%. CONCLUSIONS: Genotyping for HLA-DR by PCR-SSP is a rapid and accurate matching technique, suitable for organ transplantation, especially allocation of donor-recipients of cadaveric transplantation. PMID- 9206196 TI - [Immunohistochemical double labelling studies on liver tissues superinfected with hepatitis C virus and hepatitis B virus]. AB - OBJECTIVE: To explore the relatiship between hepatitis C virus (HCV) and hepatitis B virus (HBV) replication in liver tissue of patients superinfected with HCV and HBV. METHODS: The expresion and distribution of HCVAg and HBVAg in paraffin-embeded liver tissue from 25 autopsy cases were studied with immunohistochemical double labelling techniques using monoclonal anti-HCV NS3 and anti-HCV NS5 as well as polyclonal anti-HBs and anti-HBc. RESULTS: HBsAg and HCV NS3 or NS5 antigen were detected at the same section in 11 and 12 cases, and HBcAg and HCV NS3 or NS5 antigen in each 10 cases, respectively. Nearly all of the specimens with single labelling stained positive tissue for HC-VAg or HBVAg were also those positive with double labelling studies for HCVAg and HBVAg. The distribution of HCV and HBV infected hepatocytes was characterized as diffuse and single or cluster scattered in liver lobular. There were no differences in expression related to replication such as membranous, cytoplasmic type of HBsAg or cytoplasmic type of HBcAg and the cases positive for HCV NS3Ag or HCV NS5Ag between the group of HCV superinfected with HBV and the group infected with single HCV or HBV (chi 2 = 0.154 and 0.198, P > 0.05). CONCLUSION: The results suggested that there was no interference or suppression each other in liver tissues superinfected with HCV and HBV. PMID- 9206197 TI - [High homology between antigen of HFRSV with anti-idiotype antibody against HFRSY]. AB - OBJECTIVE: To detect the structural characterization of genes of the anti idiotype antibody and the antigen. METHODS: The hybridoma (C8) secreted the anti idiotype human McAb against hemorrhagic fever virus (HFRSV). The variable region genes of heavy and light chain of the anti-Id huMcAb were cloned. The homology was analysed between the variable region genes and the genes of HFRSV. RESULTS: The 45-55 amino acids of VH and 58-68 amino acids of VL were highly homologic to the 447-457 amino acids of G2 protein in Hantaan virus. And the homologeneous regions were similar in secondary structure of the three proteins. CONCLUSION: There is a molecular basis in the anti-idiotype antibody imaging antigen. PMID- 9206198 TI - [Invasive fungal infection in 3447 autopsy cases]. AB - OBJECTIVE: To evaluate the prevalence of invasive fungal infection at autopsy. METHOD: To retrospectively analyse 3447 autopsy cases in Peking Union Medical College Hospital from 1953 to 1993. RESULTS: There were 85 cases of invasive fungal infection at autopsy among 3447 cases. The prevalence steadily increased, especially during the recent 20 years. Among the 85 cases, only 5 patients were diagnosed clinically (5.9%). The primary diseases were mainly leukemia, cancer, and sepsis. Lung was involved in 70% of the cases, and 85% of the pathogens were fungi. CONCLUSIONS: Fungus has become a major pathogen in nosocomial infection. The prophylactic use of antifungal agents is recommended in high-risk patients because of the difficulty to establish the clinical diagnosis of invasive fungal infection. PMID- 9206199 TI - [Changes of circulating Lps and cytokines in burned patients after anti-endotoxin therapy]. AB - OBJECTIVE: Endotoxin as the inciting agent of cytokines and other mediators, whose high level expression correlates with the septic shock and MOF, has been the one of leading causes of death in ICU. METHODS: For treating sepsis and MOF caused by endotoxin, the anti-lipid A of LPS antibody was used, 19 burned patients whose TBSA varied from 50% to 100% were divided into anti-LPS treatment group and nontreated group. RESULTS: The levels of serum endotoxin, IL-6, IL-8, TNF and soluble IL-2R were lower obviously in patients of anti-LPS group than those of nontreated group (P < 0.05). CONCLUSION: Clinical study surggests that anti-lipid A of LPS antibody can act as an therapeutic agent against gram negative bacterin infection in burned patients. PMID- 9206200 TI - [The overexpression of c-met oncogene correlation with gastric mucosal lesion]. AB - OBJECTIVE: To determine whether there is a relationship between overexpression of c-met oncoprotein and stage of human gastric mucosal lesions, and its significance. METHOD: Immunohistochemical staining was used in 157 cases of endoscopic biopsies with c-met monoclonal antibody, S-19, which was raised against the human c-met gene product. RESULTS: overexpresion of c-met oncoprotein was detected in 3/30 cases (10%) of superficial gastritis, 4/33 cases (12.1%) of chronic atrophic gastritis, 10/31 cases (32.3%) of intestinal mataplasia, 10/30 cases (33.3%) of dysplasia, and 10/30 cases (33.3%) of gastric carcinoma. The positive staining rate was higher in intestinal mataplasia (54.8%), dysplasia (56.7%), carcinoma (53.3%) than in two kinds of simple chronic gastritis (P < 0.05). The positive staining was obviously located in luminal membrane of mucosal cells. The positive cells were mainly situated in proliferative cell zone of gastric glands. Moreover, the weak staining only in this zone was shown in two of the three normal mucosa. CONCLUSIONS: The overexpression of c-met may be involved in proliferation of gastric mucosa. It is possible that persistent overexpression of c-met oncoprotein is associated with the malignant transformation of gastric mucosal cells. PMID- 9206201 TI - [The effect of angelica polysaccharide on proliferation and differentiation of hematopoietic progenitor cell]. AB - OBJECTIVE: To study the effect of angelica polysaccharide (AP) on proliferation and differentiation of hematopoietic progenitor cells for clarifying the hematonic mechanism of angelica sinensis. METHODS: The techniques of culture of hematopoietic progenitor cell and hematopoietic growth factor (HGF) assay were used. RESULTS: AP could obviously promote the proliferation and differentiation of BFU-E, CFU-E, CFU-GM and CFU-MK in healthy and aniemic mice. The culture media of splenocyte, macrophage, fibroblast and skeletal muscle treated with AP had much stronger stimulating effects on hematopoietic progenitor cells. CONCLUSIONS: AP may enhance hematopoiesis by stimulating directly and/or indirectly macrophages, fibroblasts, lymphocytes in hematopoietic inductive microenvironment and muscle tissue to secrete some HGF (Epo, GM-CSF, IL, and MK-CSF). This is one of the biological mechanisms for hematonic effect of angelica sinensis. PMID- 9206202 TI - [Treating STZ-induced diabetic rats with agarose microcapsulated porcine islets]. AB - OBJECTIVE: To study the function of microencapsulated porcine islets in vitro and in vivo. METHODS: Consecutive perfusion of collagenase via pancreatic ducts was used to isolate porcine islet of langerhans. The micro encapsulates were made by phase-separation method with Chinese-made agarose as immunoisolation membrane, 21 diabetic rats were used, 6 for comparing and 15 for transplantation. Six received intraperitoneally uncapsulated islets, 6 received intraperitoneally capsulated islets, and 3 injected uncapsulated islets under kidney capsules. RESULTS: The islets could secrete insulin consecutively for a month, and the cells in capsules survived very well without autolysis. In the early two days' culture, the islets had marked reactions to the stimulation from glucose in high concentration and theophylline. The amount of insulin released was 1.83 times and 2.28 times as much as that of glucose in low concentration respectively (P < 0.01). 2000-3000 microencapsulated islets per rat were transplanted intraperitoneally in six diabetic rats without the use of immunosuppressive drugs. On the 4th day, blood sugar dropped significantly until the 7th day, when it was in normal range. The concentration of blood sugar remained for 30 days. CONCLUSION: The agarose microcapsule processes a better function of immunoisolation, which may lay a foundation of treating IDDM with encapsulated porcine islets grafting. PMID- 9206203 TI - [The effect of gamma-interferon on hepatic fibrosis in schistosomiasis japonica]. AB - OBJECTIVE: To study the effect of gamma-IFN on hepatic fibrosis in schistosomiasis japonica. METHODS: The amount and distribution of gamma-IFN and extracellular matrix, including fibronectin (FN), laminin (LN), I and II collagen which were used to judge the suppressing effect of gamma-IFN on hepatic fibrosis as markers of dynamics of extracellular matrix after administration of recombinant gamma-IFN, in liver obtained at different stages from S. japonicum infected mice were determined by immunohistochemical streptavidin biotin peroxidase complex method. RESULTS: The amount of gamma-IFN in liver reached a peak (3 mice respectively reached 2, 3 and 4 grade) at the 16th week after infection, which was higher than the levels of control groups and infected-mice at the 8-12th week (chi 2 = 15.39, 7.50, P < 0.01), and the majority of gamma-IFN was distributed round egg granuloma. FN and LN, and I and II collagen increased to 1 grade, individual 2 grade, at the 8th week after infection, which were higher than control (chi 2 = 12.44-15.39, P < 0.01), and were lineally distributed round egg granuloma. With chronicity and decrease of gamma-IFN, however, FN, LN and I, II collagen gradually increased, and reached a peak (over 70% infected mice reached 3-4 grade) respectively at the 20th week and 24th week, became wide, thick and retiform and deposited round and in egg granuloma. After administration of gamma-IFN, only 3 infected mice had 2 grade of FN at the 20th week, two mice had respectively 3 grade of type I and II collagen at the 24th week, and none of them reached 4 grade, which were significantly less than those in the untreated group at the same stage (chi 2 = 10.77, 5.05, 7.20, P < 0.01 0.05). CONCLUSION: The results indicate that gamma-IFN may play an important role in opposing the inflammatory response of egg granuloma, decreasing secretion and deposition of extracellular matrix in liver and suppressing hepatic fibrosis. PMID- 9206204 TI - [Advances in the research on new generation of oral contraceptive agents]. PMID- 9206205 TI - [Advances in the pathological classification of viral hepatitis]. PMID- 9206206 TI - [Detection of hepatitis B virus DNA and hepatitis C virus RNA in human hepatocellular carcinoma by polymerase chain reaction]. AB - In order to study the relationship between hepatitis B virus DNA (HBV DNA), hepatitis C virus RNA (HCV RNA) and liver cell carcinoma, HBV DNA and HCV RNA from the tumor tissue of 42 liver cell carcinoma cases were studied by polymerase chain reaction (PCR) and nested-PCR respectively. The results were as follows: one case of cholangiocarcinoma was positive for both HBV DNA and HCV RNA, and one case of biliary cystadenoma positive only for HBV DNA. Among the 40 cases of hepatocellular carcinoma (HCC), 29 were positive for HBV DNA and 13 positive for HCV RNA. There was no relationship obtained between HBV or HCV infection and the histological types of HCC, and HCV infection in HCC was not interrelated with HBV. It's considered that although HBV is still to be the leading cause of HCC in China, HCV may play an important role in hepatic carcinogenesis because of its high positive rate and increasing incidence. PMID- 9206207 TI - [Detection of stromelysin mRNA expression in human hepatocellular carcinoma by in situ hybridization]. AB - For the propose of detecting the significance of stromelysin (of matrix metalloproteinase family) mRNA expression in the invasion and metastasis process of liver cell carcinoma, 19 cases of human hepatocellular carcinoma (HCC) and their surrounding tissues were studied by in situ hybridization techniques. Nine cases of the HCC tissues were positive while all the tumor surrounding tissues were negative. The stromelysin expression levels were higher in those HCC complicated with portal tumor emboli or in those classified pathologically in II IV degree. It is considered that portal cancer emboli is a characteristic event for intrahepatic and extrahepatic metastasis of HCC, and matrix metalloproteinase may be of importance for the tumor invasion and metastasis. PMID- 9206208 TI - [Expression of metastasis suppressor gene nm23 in human hepatocellular carcinoma]. AB - For the purpose of investigating the relationship between the metastatic potential of the tumor as well as the expression of nm23-H1 mRNA, and for determing the location of the positive sites in the cells, tumor metastasis suppressor gene nm23-H1 in human hepatocellular carcinoma (and the nonneoplastic area surrounding the tumor) was detected by in situ hybridization using digoxiginin-labeled nm23-H1 antisense complementary RNA probe. The primary results indicated (i) positive results of in situ hybridization are presence of granules or masses in the cytoplasm; (ii) the less expression of nm23-H1 mRNA, the higher metastatic rate of the human hepatocellular carcinoma (P < 0.05); (iii) expression of nm23-H1 mRNA dose not correlate with some other factors such as tumor size and the background of other liver diseases. PMID- 9206209 TI - [The role of platelet-derived growth factor and ras P21 in experimental hepatocarcinogenesis]. AB - In order to explore whether platelet-derived growth factor (PDGF) is involved in hepatocarcinogenesis, expression of PDGF-beta chain and ras P21 were investigated using immunohistochemical method in hepatocarcinoma induced with diethylnitrosamine (DENA). Elevated PDGF-beta chain and P21 protein levels were found in hepatocytes in the early stages after DENA administration. Along with the progression of hepatocarcinogenesis, immunopositive cells were increased with the formation of various foci and nodules and the staining was usually stronger in the peripheral parts of nodules. In addition, PDGF-beta and P21 often expressed simultaneously in the smae lesions, where the cells were also positive for AFP expression. The results suggest that abnormal expression of PDGF might be an early specific event during hepatocarcinogenesis and might be involved in the malignant transformation of the hepatocytes by autocrine as well through ras P21 signal pathways. PMID- 9206210 TI - [Study on the interrelationship between human papilloma virus infection and Langerhans cell in carcinogenesis of esophagus]. AB - In order to find out the interrelationship between human papilloma virus (HPV) infection and Langerhans cells (LCs) during the development of esophageal carcinoma, and the mechanism of carcinogenesis during HPV infection, digoxigenine labelled HPV DNA probes of HPV 6B/11, HPV 16/18 with in situ hybridization, and anti-S-100 protein antibody with immunohistochemical LSAB-assays were used respectively in order to investigate HPV infection and the distribution of LCs in 40 cases of esophageal squamous cell carcinoma (ESCC). The results showed that there were fewer LC infiltration in HPV-positive cases in comparison with that of the HPV-negative cases. There were also changes about the morphology and distribution of LCs in HPV infected epithelia adjacent to the tumor. The results indicated that HPV infection might inhibit the number of locally LCs infiltrated, destroy the immune surveillance system, and work simultaneously with other carcinogenic factors, in favor of the development of ESCC. PMID- 9206211 TI - [Study on the organ weight of sudden death cases]. AB - In order to make clear any influence on the weight of various organs particularly in case of chronic diseases, Basing on the result of analysis carried out previously, data of 4402 autopsy cases of sudden death collected from 22 Medical Schools and Hospitals in China were studied. The organ weight analysed included those of the heart, lungs, spleen, liver, kidneys, brain and pancreas. The results showed that weights of the heart, lungs, spleen, liver and pancrease in males and weights of the the heart, spleen and pancreas in females were heavier than those of the non-sudden death cases in adults. The difference between male and female organ weights were that weight of the female brain aged above 3 months old and weight of all the 7 organs measured in adult female were lighter than the male. PMID- 9206212 TI - [Study on the morphometric and hemodynamic changes of the pulmonary arteries in pulmonary hypertension autopsies]. AB - With the purpose of studying the relationship between the pathological gradings and changes of the hemodynamics as well as the collagen typing of the pulmonary arteries in pulmonary hypertension, 9 autopsy cases including 6 cases of unexplained plexogenic pulmonary arteriopathy, 2 cases of diffuse interstitial fibrosis of lung and 1 case of chronic embolic pulmonary hypertension were collected as the pulmonary hypertension group (PAH). The pathological lesions of PAH in pulmonary arteries were graded as follows: Grade I showed medial hypertrophy; Grade II medial hypertropy and intimal cellular proliferation: Grade III medial hypertrophy and intimal fibrosis or embolic occlusion: Grade IV the above lesions plus plexiform lesions with/without focal necrotizing arteritis. Immunohistochemical staining with antibodies against collagen type I and type IV was performed in the paraffin sections of lungs using PAP method. The result indicated that: 1) there were significant differences between the mean medial thickness in pulmonary arteries (6.1% in the normal lung group and 26.1% in PAH) and the density of pulmonary artery (22.6% arterioles/cm2 in the normal group and 44.6 arterioles/cm2 in PAH) respectively (P < 0.01). 2) there was positive correlation between the grades of PAH lesions and the mean pulmonary arterial pressure (r = 0.68 P < 0.01). 3) collagen type I fibriles increased in the old lesions and collagen type IV fibrils were dominant in the early stage of PAH (reversible lesion). PMID- 9206213 TI - [A study on acute multiple sclerosis]. AB - With the purpose of investigating the clinicopathological changes and the criterion for differential diagnosis, ten cases of Acute multiple sclerosis (AMS) were analysed. The data suggested that its clinical course was an acute neurologic event characterized by psychic syndrome, paralysis and incontinence of bowel movement and urination. Pathologically, the demyelination lesion consisted of three patterns, i.e.: vascullar or spongy lesions, shadow plaques and liquefaction or necrosis. One of the above forms could exist individually or coexist with the others. The accurate diagnosis was made only after autopsy. Criterion for the differential diagnosis between AMS and ADEM were discussed. PMID- 9206214 TI - [An ultrastructural study of the ependymoma cells and cells of the ventric zone and neural canal of embryonic brain]. AB - In order to make clear the characteristics of cell differentiation in ependymoma, 12 ependymoma cases were studied with light and electronic microscopy, and the tumor cells were compared with cells of the ventric zone and neural canal from 8 embryonic brains. 10 out of 12 ependymoma cases were also studied with immunohistochemistry staining. According to the ultrastructural findings, four types of ependymoma were classified, namely, the primary neuroectodermal tumor with ependymoblast differentiation, the ependymoblastoma, the ependymoma, and the subependymoma. Some ultrastructural features, such as long intermediate junction and microrosette formation were considered as the important criterion for differential diagnosis,. Neuroendocrine granulas were found in one case of 12 ependymomas which is considered to be heterogenic in differentiation. PMID- 9206215 TI - [Angioimmunoblastic lymphadenopathy and angioimmunoblastic T-cell lymphoma]. AB - To investigate the clinicopathological changes of angioimmunoblastic lymphadenopathy (AILD) and angioimmunoblastic T-cell lymphoma (AITL), 5 cases of AILD and AITL were analyzed by using immunohistochemistry and Polymerase chain reaction (PCR) methods. The clinical manifestations included general lymphadenopathy, hepatosplenomegaly, fever and hematologic abnormalities. The diagnosis and differentiation of AILD and AITL were depended on the histopathologic features of lymphnodes biopsy. The presence of clusters of clear cells with variable atypia and positivity of T-cell marker were the most important diagnostic criterion for AITL. PCR analysis of TCR-beta rearrangement and EBV-genome was performed on 4 of 5 cases. All of the cases showed clonal rearrangement of TCR-beta and 3 of them were EBV-DNA positive. The results suggest that AILD might be a prelymphomatous lesion, related to EBV infection with a high incidence of developing to malignant lymphoma. PMID- 9206216 TI - [A pathological survey of the therapeutic effect on experimental hypoxic-ischemic encephalopathy]. AB - Rat models of the acute and recuperative phases of hypoxic ischemic encephalopathy (HIE) were established beginning by the 7th day after birth through ischemia and hypoxia. The prophylatic and therapeutic effects on experimental HIE were studied by the application of radix salviae miltiorrhizae, flunarizine and hyperbaric oxygen. Experimental data indicated that among these measures, radix salviae miltiorrhizae gave a better result and the pathological change in the prophylactic and therapeutic groups particularly the result of the latter one were light serious than those of the control group. PMID- 9206217 TI - [Advances in the study on the inhibition of vascular smooth muscle proliferation by gene rearrangement and gene transfer techniques]. PMID- 9206218 TI - [Problems in the application of cataract extraction with phacoemulsification aspiration]. PMID- 9206219 TI - Phacoemulsification and posterior chamber intraocular lens implantation. AB - OBJECTIVE: The study was designed to evaluate the therapeutic effects of phacoemulsification and PMMA posterior chamber intraocular lens implantation. METHODS: The surgery was performed on 74 eyes of 67 patients with senile, complicated and congenital cataracts. RESULTS: Post-operatively, the visual acuities with spherical correction or without correction were 0.5 or better in 80.5% at one week and with correction, in 97.6% at three months. The mean astigmatism was 1.90 +/- 1.15 D at one week and 0.93 +/- 0.55 D at three months which were respectively less than that in the control group with a 10-12 mm large incision (P < 0.001), but almost the same as that in the control group with a small incision and manual nucleus division technique (P > 0.05). The main complications were iris bite in 6 eyes, capsular rupture in 2 eyes and aseptic hypopyon in one eye. The selection of cases, surgical techniques and management of intraoperative complications were discussed. CONCLUSION: These results suggest that phacoemulsification be applied extensively. However, that it has more complications and risk must be emphasized for a beginner. PMID- 9206220 TI - [Cataract extraction by phacoemulsification using in situ nuclear fracture technique]. AB - OBJECTIVE: The study was designed to investigate the technique of phacoemulsification in which the lens nucleus is fractured without collapse of nucleus into anterior chamber. METHODS: Phacoemulsification using in situ nuclear fracture technique with posterior chamber intraocular lens implantation was performed on 55 eyes of 52 patients with senile cataract. RESULTS: The in situ nuclear fracture technique was successfully completed in 92.72% of all the eyes: the average time of lens phacoemulsification was 2.26 minutes, the rate of vitreous loss was 5.46%, the rate of corneal endothelial cell loss was 12.13%, and one week after the operation, the visual acuities of 0.5 or better were obtained in 72.72% of the cases. CONCLUSION: The key points of in situ fracture technique by phacoemulsification include selection of softer nucleus, continuous ring-shaped capsulorhexis, complete hydrodissection and fracture of the nucleus quadrant by quadrant. PMID- 9206221 TI - [Phacoemulsification aspiration and intraocular lens implantation]. AB - OBJECTIVE: To investigate the characteristics of phacoemulsification of cataract in our country and management and prevention of its complications. METHODS: The procedure of phacoemulsification with small incision (6 mm) intraocular lens implantation was improved and was performed on 40 eyes of 40 cases. They had been followed up for 3-6 months postoperatively and their follow-up results were summarized. RESULTS: The uncorrected visual acuity 1.0 or better was in 57.5%, 0.8 or better in 82.5% and 0.5 or better in 97.5%. The postoperative mean astigmatism was 0.50 +/- 0.25 D, which was less than that in the large incision group (9-12 mm) performed on 40 cases in the same period (P < 0.001). Cataract with hard nucleus can be successfully taken out by the technique of peripheral rotation engraving method. CONCLUSION: The operation is a safe reliable one for restoration of visual acuity in patients with cataract, and is superior to extracapsular cataract extraction. PMID- 9206222 TI - [Accommodation in pseudophakic eyes with monofocal posterior chamber intraocular lens]. AB - OBJECTIVE: To study the accommodative problem of pseudophakic eyes with commonly implanted monofocal posterior chamber intraocular lens (PC-IOL). METHODS: Slataper's method, dynamic retinoscopy and pattern-visual evoked potentials were used to measure the accommodative power in 120 pseudophakic eyes (101 patients) with monofocal PC-IOL. RESULTS: The mean accommodative power was 2.15 +/- 0.76 D. Stepwise regression analyses revealed that the apparent accommodation is positively correlated with the corrected distant vision and the distantly corrected near vision, respectively. CONCLUSION: The apparent accommodation of pseudophakic eyes does exist, but it is a false one. The result of accommodative power measured by dynamic retinoscopy is the most objective and precise. The central visual efficiency and the applicable range of accommodation in pseudophakic eyes with low myopia (-1.00 D) are better than that in emmetropic pseudophakic eyes. PMID- 9206223 TI - [Ultrastructural changes in various cataractous lenses in rats]. AB - OBJECTIVE: To study the ultrastructural changes of lenses aged 50 days from normal rats and rats with three kinds of cataract. METHOD: Transmission electron microscope (TEM) was used to investigate the ultrastructural changes of normal lens, selenium, galactosemic and congenital cataracts in the rat. RESULTS: In the normal group, the structure of cell membrane in cortex is not changed obviously, but in the lens nucleus there are slight cellular degeneration, low cytoplasmic density and the destruction of intercellular junctional complexes. In the selenium cataratous lens, in the lens cortex and nucleus the cellular junctional complexes are damaged severely, the cystic degeneration of mitochondria in the cytoplasm is obvious, the cytoplasmic density becomes lower and slight liquefaction can be observed in some parts of cytoplasm. In the galactosemic cataractous lens, in the cortex obvious liquefaction, vesicles and globular structures can be found within the cytoplasm, intercellular spaces are enlarged and liquified markedly, and in the lens nuclear area the cytoplasm appears as fine sands. In congenital cataractous lens, the cellular structure in cortex is essentially normal, but there are marked destruction of junctional complexes, obvious cystic degeneration of mitochondria and cytoplasmic aggregation with uneven distribution in the lens nuclear area. CONCLUSION: The ultrastructural changes in different kinds of cataractous lenses depend on the extent and location of opacification in lenses. PMID- 9206224 TI - [Promotion of c-fos oncogene expression in cultured rabbit lens epithelial cells by macrophages]. AB - OBJECTIVE: This study was designed to test the hypothesis that inflammatory cells deposited on the surface of implanted intraocular lens stimulate the proliferation of residual lens epithelial cells. METHODS: Immunocytochemical ABC methods with purified anti-human Fos protein serum antibody was used to stain the cultured rabbit lens epithelial cells incubated with rabbit macrophages and macrophage-conditioned medium (MCM). RESULTS: The lens epithelial cell growth was faster in the cultures with macrophages and MCM than in the controls. Fos protein positive staining was found in the nuclei of epithelial cells cultured with macrophages for 4 hours, and of the cells with MCM for one hour; whereas the staining was negative in the cells of control cultures. CONCLUSION: Macrophages promote c-fos oncogene expression in the lens epithelial cells, that might be mediated by bioactive factors secreted by macrophages. PMID- 9206225 TI - [A study on cAMP and Ca-CaM of the epithelium in human normal and cataractous lenses]. AB - OBJECTIVE: The study was designed to study the relationship between calcium calmodulin (Ca-CaM) and cyclic adenosine monophosphate (cAMP) systems and their possible cataractogenic effects. METHODS: We measured cAMP, Ca and Ca-CaM in the epithelium of human normal and cataractous lenses by enzymic and radioimmunoassay methods. RESULTS: The levels of Ca-CaM in the epithelium of senile cataractous lens were higher, while of cAMP were lower than that in the normal lens. CONCLUSION: The results imply that the low level of cAMP might cause the high level of calcium in the lens which is possibly a cataractogenic factor. The dangerous role of calcium is amplified by the elevation of Ca-CaM. PMID- 9206226 TI - [The changes of hemodynamics in ocular trauma and treatment with compound anisodine]. AB - OBJECTIVE: To study the changes of hemodynamics in human ocular trauma and treatment with compound anisodine. METHODS: 34 cases with ocular trauma were examined by rheoophthalmography (ROG) and Doppler ultrasonography (DUSG). RESULTS: It appeared that the wave amplitudes of ROG were lower in both injured and contralateral eyes, which indicated the ocular blood flow was reduced. The speed of systolic blood flow in ophthalmic artery (OA), the value of systolic peak frequency (PK) in internal carotid artery (ICA) and common carotid artery (CCA) in DUSG were increased, indicating that there were certain degrees of blood vessel spasm in OA, ICA and CCA. CONCLUSION: Ocular trauma not only directly impairs the vasomotor function of injured eyes but also affects the vasomotor function of contralateral eyes and the function of the autonomic nervous system or cerebral cortex by consensual reaction. The treatment by compound anisodine may effectively relieve and restore the above changes and improve the visual function. ROG and DUSG are sensitive and objective to estimate the changes of hemodynamics and evaluate the therapeutic effect in ocular trauma. PMID- 9206228 TI - [Treatment of neovascular glaucoma by anterior retinal cryotherapy and trabeculectomy]. AB - OBJECTIVE: To investigate the efficacy of anterior retinal cryotherapy (ARC) for treatment of neovascular glaucoma (NVG). METHODS: 12 eyes (11 patients) with open angle NVG were treated by ARC only, 22 eyes (22 patients) with closed angle NVG were treated by ARC combined with trabeculectomy, and 32 eyes (31 patients) with closed angle NVG were treated by either ARC or cyclocryotherapy as controls. The follow-up period were 6-26 months. RESULTS: Iris new vessels (INV) regressed or disappeared with normal intraocular pressure (IOP) in 92% (11/12) of the eyes with open angle NVG treated simply by ARC. In comparison with closed angle NVG treated by the same method, there was a significant difference in IOP (P < 0.05). The combination of ARC and trabeculectomy showed markedly better results than either ARC or cyclocryotherapy for treatment of closed angle NVG (P < 0.01). In the eyes with closed angle NVG, INV regressed or disappeared with normal IOP in 86% (19/22), 90% (18/20) achieved a marked relief from pain and the visual acuity was better or unchanged in 67% (8/12) that was significantly different from the eyes treated by cyclocryotherapy (P < 0.05). CONCLUSION: The results indicate that ARC is suitable for treatment of NVG at early stage, and ARC combined with trabeculectomy, at later stage. PMID- 9206227 TI - [A study of neuroretinal rim morphology in physiologic large cups and early glaucoma]. AB - OBJECTIVE: To find the differences of the physiologic large cups from the optic cup of early glaucoma. METHODS: The morphology of neuroretinal rim was studied. The subjects were divided into two groups: (1) 54 cases 88 eyes with physiologic large cups defined by cup/disk (C/D) ratio more than 0.6, optic disk areas more than 2.8 mm2, without neuroretinal rim loss within 3-6 years of follow-up, normal intraocular pressure and visual field. (2) 68 cases 89 eyes with early glaucoma defined by visual field defects, neuroretinal rim loss in follow-up and C/D ratio less than 0.8. The indexes of neuroretinal rim related to morphology are a series of rim widths and a parameter of cupping morphology which is a ratio of the vertical C/D over the horizontal C/D. RESULTS: The characteristics of a physiologic large cup are a large optic disk, a horizontal elliptic cupping and the widest neuroretinal rim at the inferior, followed by superior, nasal and temporal, while the characteristics of optic nerve head of glaucoma are normal size of optic disk, vertical elliptic cupping and the widest rim not at the inferior, since the inferior rim is subject to glaucomatous damage. CONCLUSION: To differentiate a physiologic large cup from a glaucomatous one, the most effective way is to apply the parameter of cupping morphology in combination with neuroretinal rim area and optic disk area in the multivariate discrimination. PMID- 9206229 TI - [Endolaser photocoagulation in vitrectomy surgery]. AB - OBJECTIVE: To verify the usefulness and effectiveness of endolaser photocoagulation in vitrectomy. METHODS: Endolaser photocoagulation was applied in 40 cases (45 eyes) of vitrectomy surgery. Indications for vitrectomy included vitreous hemorrhage 31 eyes (proliferative diabetic retinopathy 13, Eales's disease 13, retinal vein occlusion 4, and scleral penetrating injury with intraocular foreign body removal 1), complex retinal detachment 12 eyes, intraocular nonmagnetic foreign body removal and postoperative silicone oil tamponade 1 eye respectively. Treatment involved pan-retinal scatter therapy in 7 eyes, focal therapy in 22 eyes, sealing primary or iatrogenic retinal breaks in 14 eyes, draining retinotomy in 2 eyes, and encircling scleral buckle photocoagulation in 3 eyes. The follow-up times ranged from 2 to 18 months (average 9 months). RESULTS: Whitening of laser spots was not apparent in 6 eyes with retinal breaks following incomplete gas-fluid exchange. Recurrent retinal detachments occurred in 5 eyes postoperatively. CONCLUSION: Endolaser photocoagulation facilitates the performance and reduces the complications of vitrectomy surgery. PMID- 9206230 TI - [A clinical report on mixed astigmatism]. AB - OBJECTIVE: This study was designed to analyze the common cause of asthenopia, mixed astigmatism. METHODS: The types and characters of mixed astigmatism in 356 eyes (207 patients) were analyzed and their naked and corrected visual acuities were investigated. Some problems of mixed astigmatism were discussed. RESULTS: Most of the patients had evident visual defects, the average naked visual acuity being 0.3 (20/60). However, all the patients had better corrected visual acuities with an average of 0.6 (20/30). CONCLUSIONS: Due to the presence of accommodation during the examination, an incorrect lens may be prescribed. Three mistakes might occur, (1) A mixed astigmatism is not detected, and a simple myopic lens is prescribed; (2) An eye with mixed astigmatism is diagnosed as simple or compound myopic astigmatism; (3) A hyperopic astigmatism is diagnosed as mixed astigmatism. The visual defects caused by mixed astigmatism can be satisfactorily corrected by spectacles. PMID- 9206231 TI - [Surgical management for severe complications following radial keratotomy]. AB - OBJECTIVE: To investigate the surgical treatment for severe complications after radial keratotomy (RK). METHODS: A protocol of surgical treatment for 3 severe complications after RK, i.e., early corneal perforation and infection, corneal rupture following contusion and corneal leucoma, is proposed based on modern microsurgical techniques and computer assisted corneal topography. RESULTS: All eyes with such severe complications were salvaged and their visual outcomes were also satisfactory. CONCLUSIONS: Severe complications after RK threaten the eye. Special management should be applied for these situations. With the help of corneal topography, the refractive results of corneal surgeries can be improved. PMID- 9206232 TI - [Relationship between cellular DNA and expression of epidermal growth factor receptor in pleomorphic adenoma of lacrimal gland]. AB - OBJECTIVE: To obtain the information about whether expression of epidermal growth factor (EGF) receptor is related to the proliferative activity of tumor. METHODS: The expression of EGF receptor and DNA content of pleomorphic adenoma of lacrimal gland in 32 cases were detected by an immunohistochemical avidin biotin peroxidase complex method and determined by an image analysis technique, respectively. The relationship between the expression of EGF receptor and DNA content was analyzed. RESULTS: 10 tumors were stained positively with anti-EGF receptor antibody, the DNA contents of 14 cases were increased, their DNA ploidy distribution pattern showed two or several peaks. Good correlation has been found between the expression of EGF receptor and DNA ploidy distribution pattern, and the DNA distribution pattern of tumor with positive EGF receptor expression showed mainly two or several peaks (P < 0.01). CONCLUSION: The expression of EGF receptor of pleomorphic tumor of lacrimal gland is related to the proliferative activity of tumor cells. PMID- 9206233 TI - [Establishment of in vitro culture of bovine trabecular meshwork cells and their phagocytosis]. AB - OBJECTIVE: To study the role of abnormality of phagocytosis of trabecular meshwork cells in the pathogenesis of glaucoma. METHOD: In vitro trabecular meshwork cell culture was successfully established from excised bovine aqueous outflow pathway. Using latex microspheres 0.6 micron in diameter as markers, we investigated the phagocytosis of cultured bovine trabecular cells and its influential factors qualitatively and quantitatively. RESULTS: Immunohistochemical staining studies have shown that trabecular cells secrete fibronectin and laminin. It is shown that bovine trabecular cells in vitro had strong phagocytic ability to ingest the particles, the number of latex beads in cells was increased with the prolongation of time in culture, and by 24 hours of incubation, the average number of beads in each cell was 40.5 +/- 6.7, in the mean time marked morphological and damaging changes, even death, of trabecular cells occurred. Epinephrine and cortisone inhibited significantly the phagocytosis of cells (P < 0.05), while epidermal growth factor had no such effect. CONCLUSION: The phagocytosis of trabecular cells is important to keep aqueous humour outflow patent, and abnormal phagocytosis of trabecular cells might be related to the pathogenesis of primary glaucoma. PMID- 9206234 TI - [A study on changes of levels of products of lipid peroxidation in retina of rat with experimental autoimmune uveoretinitis]. AB - OBJECTIVE: To understand the mechanisms of the retinal tissue destruction of rat with experimental autommune uveoretinitis (EAU) induced by rabbit retinal soluble antigen. METHOD: The levels of the following products of lipid peroxidation were determined at different times, conjugated dienes, malondialdehyde, fluorochrome lipid and lipid hydrogen peroxide. RESULTS: In the process of EAU, the levels of products of lipid peroxidation in the retina were increased gradually in various degrees. CONCLUSION: It is suggested that the damage from lipid peroxidation mediated by free radicals of the retina be one of the factors which induces the retinal destruction in EAU. PMID- 9206235 TI - [An experimental study on corneal collagen shield vehicle for delivery of pilocarpine]. AB - OBJECTIVE: The study was designed to clarify the role of corneal collagen shield made in China as a drug vehicle for the delivery of pilocarpine in different ways. METHODS: 80 eyes of New Zealand white rabbits were randomly divided into three groups, corneal collagen shield immersed with pilocarpine, corneal collagen shield with topical application of pilocarpine and synthetic collagen pilocarpine shield. At 0.5, 1, 3 and 6 hours after the delivery, the aqueous concentrations of pilocarpine and the pupillary sizes of all the eyes were measured. RESULTS: This study showed that shortly after the delivery of pilocarpine in the 3 groups, in spite of difference in ways of its delivery, its aqueous concentrations reached a relatively high level and maintained for more than six hours, the shield acting as a temporary "drug source". The aqueous pharmacokinetics of corneal collagen shield for pilocarpine delivery was consistent with the first order kinetics and it was confirmed by the observations of miotic response. CONCLUSIONS: The corneal collagen shield made in China is a fine vehicle for drug delivery. PMID- 9206236 TI - [A study on neural regeneration after experimental epikeratophakia]. AB - OBJECTIVE: To assess the effect of epikeratophakia (EKP) on corneal nerve regeneration and the recovery of corneal sensitivity. METHODS: 36 adult rabbits underwent epikeratophakia with either 0.2 mm or 0.3-0.35 mm circular trephination. A histochemical technique (acetylcholine esterase staining) was used to investigate the process of corneal nerve regeneration, and the recovery of corneal sensitivity was determined in 36 rabbits at various intervals after EKP. RESULTS: The corneal sensory nerves began to regenerate just peripheral to the host-donor interface 3 weeks following EKP, the nerve fibers traveled toward the stroma of the lenticules at 2 months, and formed a sparse subepithelial plexus at 8 months after surgery. A relative hyposthesia was found in the central zone of the lenticule when compared with its peripheral zone 4 months postoperatively. The sensitivity of the central zone remained lower at 5-8 months after EKP though it had remarkably increased. The density of re-innervated fibers was higher in the shallow incision than that in deeper one. CONCLUSION: It is suggested that the factors affecting the corneal innervation and the recovery of corneal sensitivity following EKP include; (1) the scar formation as a barrier; (2) the adaptation to the micro-environment around the lenticule; and (3) the depth of trephine cut. Therefore, a 0.2-0.25 mm depth of trephine incision is advantageous to the lamellar dissection, graft suturation and corneal nerve regeneration. PMID- 9206237 TI - [Prospects on the development of traditional Chinese drugs in the new century]. PMID- 9206238 TI - [Study on effect of strengthening body resistance method on asthmatic attack]. AB - In order to study the effect of strengthening body resistance method on asthmatic attack, asthma patients of Cold type and Heat type were treated with symptomatic treatment (as control groups), and treatment both on the principal and secondary aspect (as test group) respectively. The results showed that the markedly effective rates of the test groups of both type were higher than that of the two control groups. In test groups after treatment, the 1 second forced expiratory volume (FEV1) and the maximal expiratory flow increased markedly, the human leucocyte antigen carrying (HLA-DR+) T cell ratio, the basophilic cell releasing capacity (HBR) and the T cell hyperplasia response to specific allergen were all reduced. While in the control groups, these criteria had no significant changes after treatment. These results suggested that the strengthening body resistance method displayed an obvious regulating action in relieving attack of asthma. PMID- 9206239 TI - [Clinical observation of wenxin decoction in treating 82 patients with spontaneous angina pectoris]. AB - One hundred twenty-two patients with spontaneous angina pectoris (SAP) were randomly divided into treated group (82 cases) and control group (40 cases), and treated with Wenxin decoction and isosorbide dinitrate respectively. Results showed that in treated group the total effective rate of SAP was 95.12% and that of electrocardiographic findings were 74.39%. These results were all superior to those of control group (P < 0.01). Moreover, results of extracorporeal thrombosis test showed after Wenxin decoction treatment, the length of thrombus decreased from 23.56 +/- 5.47 mm to 20.04 +/- 5.17 mm, the wet weight of it decreased from 92.65 +/- 18.45 mg to 76.94 +/- 15.08 mg and the dry weight from 21.76 +/- 7.30 mg to 16.90 +/- 5.35 mg. The submaximal exercise test revealed an increase of exercise time from 474.66 +/- 96.33 seconds to 548.83 +/- 99.93 seconds, increase of acting quantity from 104.16 +/- 19.65 W to 123.61 +/- 24.96 W and a decrease of ST segment depression from 0.183 +/- 0.041 mV to 0.139 +/- 0.038 mV. These results suggested that Wenxin decoction is valuable in treating SAP. PMID- 9206240 TI - [A clinical study of hehuantang in treating coronary heart disease]. AB - Fifty-nine cases with angina pectoris (AP) in coronary heart disease were divided randomly into two groups, 37 cases of Hehuantang group (HT) treated with Hehuantang, 22 cases of patient treated with nifedipine tablet was taken as control group (CG). THE RESULTS: (1) the marked effective and total effective rate in allevating AP was 75.68% and 91.89% in HT respectively; (2) activities of serum SOD and whole blood GSH-Px in HT significantly elevated than that pretreatmentally (P < 0.01); (3) comparing with CG, SOD activity and GSH-Px/LPO ratio increased (P < 0.01 and P < 0.05), whereas plasma LPO content lowered (P < 0.05). It suggested that HT having effects of relieve AP and enhance antioxidative activity and attenuate lipid peroxide reaction. The mechanism might be correlated to inhibit the calcium overload and reduce LPO production in cardiac cell as well as improve blood supply to myocardial ischemia. PMID- 9206241 TI - [Clinical study of zhinu-I,-II in treating 61 patients with idiopathic thrombocytopenic purpura]. AB - The article studied 61 patients with idiopathic thrombocytopenic purpura, they were classified and treated with Chinese herbal medicine Zhinu (ZN) -I, -II, in comparison with 30 cases of patients treated with prednisone. The results showed that the effect of prednisone was more effective than ZN- I, -II in four weeks (P < 0.05). On the contrary, follow up for 12 months, the effect of ZN-I, -II were more effective than prednisone (P < 0.05) and the possibility of recurrence was lower (P < 0.01). In addition, blood platelet counts of the patients treated with ZN-I, -II increased slower, but lasting for longer term. The conclusion indicated that the ZN-I, -II were safe and effective. There was no obvious side effects. PMID- 9206242 TI - [Clinical and experimental study on effect of ganyan IV in treatment of chronic active hepatitis complicated with hyperbilirubinemia]. AB - Sixty chronic active hepatitis patients complicated with hyperbilirubinemia (total bilirubin > 171 mumol/L) were treated with combined treatment of Ganyan IV and Western medicine. The curative effect was compared with that treated with Western medicine alone as control (56 cases). Result showed that the effect of combined therapy group was much better than that of the control in eliminating the jaundice, descending the alanine transaminase (ALT) and improving the reversed A/G ratio (P < 0.05-0.001). In experimental studies, Ganyan IV was applied to the mice with acute liver damage formed by CCl4. It also showed significant effect on reducing total bilirubin and elevating the serum albumin statistically as compared with control (P < 0.05 = 0.01). In addition Ganyan IV could accellerating the bile excretion of normal as well as of liver damaged rats significantly. It was concluded that the Ganyan IV has the effects of treating jaundice, descending transaminase, elevating serum albumin and improving A/G ratio. PMID- 9206243 TI - [Study on mechanism of eye-signs in blood stasis syndrome]. AB - Through a study of 504 cases of observation group with eye-signs in blood stasis syndrome (BSS) and 112 cases of control group without eye-signs in BSS, it has been found that in the observation group, the scores of the blood concentration, viscosity, aggregability and coagulability, level of plasma thromboxane B2 (TXB2), and the ratio of TXB2/6-keto-PGF1 alpha were obviously higher than those in the control group, but level of 6-keto-PGF1 alpha was obviously lower than that in the control group; the above-mentioned parameters of blood hyperviscosity syndrome, was obviously higher than that in the control group (90.08%:2.68%); comparisons between the two groups were significantly different (P < 0.001). Certain findings of the pathological base of eye-signs in BSS were found in the investigations. PMID- 9206244 TI - [Relation between the blood stasis syndrome of hepatopathy and hemodynamics of portal vein]. PMID- 9206245 TI - [Experimental studies on inhibitory effects of radix Peucedani on hypoxic pulmonary hypertension in rats]. AB - Light microscopic and morphometric methods along with the measurement of right ventricular systolic pressure (RVSP) and right ventricle hypertrophy index (RBHI) were performed by the authors in studying the inhibitory effects of Radix Peucedani (RP) on structural remodeling of intraacinar pulmonary arteries (IAPA) and pulmonary hypertension (PHT) in hypoxic rats. The results showed that, RP could not only antagonize IAPA contraction caused by hypoxia and lower the resistance of pulmonary circulation, inhibit the cell proliferation and hypertrophy of adventitial cells of pulmonary artery, but also reverse the change in structure and function of pulmonary artery. It suggests that RA plays a key role in inhibiting hypoxic structural remodeling of IAPA and pulmonary hypertension. PMID- 9206246 TI - [Effect of jiawei sijunzi decoction on migrating myoelectric complex in 8 Gy irradiated rats]. AB - The normal intestinal migrating myoelectric complex (MMC) of rats recorded by implanted electrode consists of four phases (phase I, II, III and IV). After 8 Gy of gamma-radiation for 1 hour to 7 days, the MMC cycle in most of the rats were disappeared only phase I or II existed with minute's rhythm. 1 hour or 3 days after radiation, the MMC cycle appeared in a few rats with the phase II shortened significantly (P < 0.05). Results of observation on effect of Jiawei Sijunzi Decoction on MMC after radiation showed the changed phase and cycle of MMC were normalized basically by the medication. These results suggested that the Jiawei Sijunzi decoction could improve the intestinal disturbances caused by radiation, it might be one of the reason of its alleviating effect on the radiation diarrhea. PMID- 9206248 TI - [Research and create new drugs by integrated traditional Chinese medicine and Western medicine]. PMID- 9206247 TI - [Study on the enhancing effect of polyporus polysaccharide, mycobacterium polysaccharide and lentinan on lymphokine-activated killer cell activity in vitro]. AB - The actions of Polyporus polysaccharide (PPS), mycobacterium polysaccharide (MPS) and lentinan (LEN) on lymphokine-activated killer (LAK) cell activity in vitro were investigated in this study. Human peripheral blood mononuclear cells (PBMC) were cultured for 96 hours with medium containing different concentrations of the above-mentioned drugs in combination with recombinant interleukin 2 (rIL-2). Then cell-mediated lysis was determined by 1H-TdR release assay including NK sensitive and NK resistant target cells. The results demonstrated that, when combined with rIL-2 in a certain concentration, all three kinds of polysaccharides could enhance the LAK activity by 42%-56.9%, and reduce the dose of rIL-2 by 50% (P < 0.05-0.01). It suggested that the PPS, MPS and LEN could be used as bioactivity regulators in LAK cell therapy in tumor treatment. PMID- 9206250 TI - [Modern research on the types of hypertension in traditional Chinese medicine]. PMID- 9206249 TI - [Review on the research of reinforcing body resistance and removing stasis compound kangshou yin in postponing senility]. PMID- 9206251 TI - [Textual studies on shangzhou zhiqiao fructus Aurantii]. AB - The original plant of Shangzhou Zhiqiao, the best among the species of Zhiqino, is confirmed as Citrus aurantium rather than Poncirus trifolia in this paper based on the description and drawings of Zhiqiao in (Bencao Gangmu) by Li Shizhen in the 16th rentury, samples from Shangzhou(Shanxi Province), and the cold-warm period of historical climate in the local area, which is believed to be related to the distribution of C. aurantium in Shangzhou. However, P.trifolia has often been confused with C. aurantium by some herbalists. PMID- 9206252 TI - [A comparative study on amino acids of guang horn, buffalo horn and cattle horn]. AB - The results show that the total amino acids of three species of horns are different to a certain degree, but the composition and content distribution of these acids are similar. The composition and content of different parts of two species of ox horns are similar too. PMID- 9206253 TI - [Pharmacognostical identification of fructus Meliae azedarch]. PMID- 9206254 TI - [Effects of processing on specific toxicity and pharmacodynamics of radix Kansui, radix Achyranthis bidentatae and semen Armeniacae amarum]. AB - The comparative toxicological researches on crude and processed drugs show that the activating action of Radix Kansui, Radix Achyranthis Bidentatae and Semen Armeniacae Amarum on EBV-EA can be decreased by processing. Processing can also decrease stimulating activity on mouse skin, inhibit tumor-promoting activity in two stage skin tumor promoting test and lapactic effect by Radix Kansui. Meanwhile the pharmacological effects of these drugs can be retained or increased by processing. PMID- 9206255 TI - [Effect of ginger-processing on l-ephedrine contents in rhizoma Pinelliae]. AB - The contents of l-ephedrine in Rhizoma Pinelliae have been determined to be approximately 3.44 x 10(-3). The contents vary with the five different processing methods in the following order, Pinellia boiled with ginger juice and alum > raw Pinellia > Pinellia dipped in ginger juice > Pinellia boiled only with ginger juice > Pinellia dipped in alum solution. The alum solution and the Pinellia boiled only with ginger juice bear most strongly on the l-ephedrine contents of Pinellia. PMID- 9206256 TI - [Extraction technology of effervescent granules for arresting cold pain optimized by orthogonal tests]. AB - In the extraction technology of Effervescent Granules for Arresting Cold Pain (Hantongding Paoteng Chongji), the drug combinative modes, solvent pH and drug granularity were optimized by orthogonal tests, with the total alkaloid, paeoniflorin and glycyrrhetinic acid as indexes. The experimental results show that it is better to decoct together all the recipe ingredients, with water of pH2 for the first decoction, then water of pH8 for the second decoction, and to make its granularities ranging from the original herbal pieces to particles which can pass through a No. 2 sieve. PMID- 9206257 TI - [Flavone constituents in the seeds of Sophora vicii folia Hance]. AB - Four flavone compounds have been separated from the mature seeds of Sophora vicii folia. By means of physico-chemical and spectroscopic analysis, their structures have been identified as 5,7,3'-trihydroxy-4'-methoxyflavone (I); 7,3'-dihydroxy 1'-methoxyflavone (II); 7,4'-dihydroxyflavone (III) and 7,3',4'-trihydroxyflavone (IV). All of them are found in S. viciifolia for the first time, and I and II are found Sophora genus for the first time. PMID- 9206258 TI - [Chemical constituents of the fruits of Vitex trifolia L]. AB - Five compounds were isolated from the fruits of Vitex trifolia and identified on the basis of spectral data and chemical methods to be rho-hydroxybenzoic acid, beta-sitosterol, beta-sitosterol-3-O-glucoside, casticin and, 3,6,7 trimethylquercetagetin. Except beta-sitosterol all of the compounds were obtained from the fruits of the plant for the first time. PMID- 9206259 TI - [GC-MS analysis of essential oil from pericarp of Illicium modestum A. C. Smith]. AB - The chemical constituents of essential oil from the pericarp of Illicium modestum were analyzed and 60 compounds were identified by GC-MS. Among them anethole (main compound in the oil of Chinese anise star) and safrole were absent. PMID- 9206261 TI - [Interaction of human serum albumin with berberine hydrochloride by fluorescence method]. AB - Fluorescence method has been used to observe the fluorescence quenching of human serum albumin (HSA) by its interaction with berberine hydrochloride (BH). The interaction dissociation constants of HSA and BH have been determined from a double reciprocal Lineweaver-Burk plot (Kd = 1.73 x 10(5) mol/L). PMID- 9206260 TI - [Antitussive, expectorant and anti-inflammatory effects of rhizoma Cynanchi stauntonii]. AB - Experiments indicated that the ethanol extract and ether extract from Rhizoma Cynanchi Stauntonii showed obvious antitussive and expectorant effects in mice with stomach administration. The antitussive effect of ethanol extract was stronger than the expectorant effect. The water extract showed a certain expectorant effect but no marked antitussive effect with oral administration. Obvious anti-inflammatory effect was observed on croton oil-caused mouse ear swell with intraperitoneal injection. PMID- 9206262 TI - [Inhibitory effect of tetrandrine on collagen synthesis of experimental hepatic fibrosis in rats]. PMID- 9206264 TI - [Effect of dl-3-n-butylphthalide on delayed neuronal damage after focal cerebral ischemia and intrasynaptosomes calcium in rats]. AB - Effect of dl-3-n-butylphthalide on the size of infarction and behavior changes were investigated after delayed neuronal damage in rats subjected to permanent middle cerebral artery occlusion (MCAO) by the method of Tamura et al, and the scores of behavior were evaluated by the method of Bederson et al. The results show that the size of infarct area was significantly reduced 2 h after MCAO following administration of NBP at the dose of 80, 160 and 240 mg.kg-1, the percentage of reduction of infarct area was 49.0%, 69.5% and 85.1% respectively. Neurological deficit was also improved. The size of infarction and the score of neurological deficit were also reduced significantly following pretreatment with NBP at the dose of 80 mg.kg-1 per day for 7 days by the end of the final dosage 24 h before MCAO or at the single dose of 160 mg.kg-1 1 h before MCAO. The results suggest that NBP has therapeutic and preventive effect on stroke and imply that NBP has the action of attenuating neuronal damage after delayed cerebral injury. In addition, the level of calcium ([Ca2+]i) in rat intrasynaptosomes was determined using fura-2 (a fluorescence indicator) technique. It was found that NBP can not lower the rise of [Ca2+]i induced by 30 mmol.L-1 KCl. However, the effect of NBP on [Ca2+]i overload induced by excitatory amino acid remains to be studied. PMID- 9206263 TI - [Protective effect of tetrandrine on neuronal injury in cultured cortical neurons of rats]. AB - The ability of tetrandrine (Tet), as a Ca2+ antagonist isolated from a traditional Chinese herb, to reduce cortical neuronal injury was quantitatively examined in cell cultures derived from fetal rats by measurement of lactate dehydrogenase (LDH) released to the extracellular bathing media. Cell cultures exposed to excitotoxins-glutamate (Glu), N-methyl-D-aspartate (NMDA), beta-N oxalylamino-L-alanine(BOAA, on non-NMDA receptors) and beta-N-methylamino-L alanine(BMAA, on NMDA receptors)-for 24 h showed widespread neuronal injury, which was substantially attenuated by addition of Tet 10(-7)-10(-6) mol.L-1 except to NMDA. Tet failed to protect neurons against NMDA. These results suggest that Tet has protective effect on fetal rat cortical neuronal injury induced by some excitotoxins in vitro. The mechanism of action was hypothesized that opening of Ca2+ channel in cellular membrane would not happen, because of inhibition of Na+ influx and membrane depolarization induced by Tet. As a result, cytosolic free Ca2+ overload and then neuroal injury were prevented or lightened. PMID- 9206265 TI - [Antitumor activity of yungumycin]. AB - Yungumycin, produced by a Streptomyces strain which was isolated from a soil sample collected in Guanping Nature Conservation Zone, Yunnan Province, China, has been verified to be identical with gougerotin. Determined by clonogenic assay, the IC50 of yungumycin to KB cells was found to be 1 microgram.ml-1. By spermatogonial assay, the activity of yungumycin was very close to that of 5-FU and MTX. Administered by i.p. route, yungumycin showed moderate inhibition against colon carcinoma 26 in mice. However, yungumycin by oral administration exerted highly inhibitory effects on both colon carcinoma 26 and sarcoma 180 (solid tumor) in mice and the inhibition rates reached 85% and 83%, respectively, at tolerable dose. Compared at equitoxic dose of 1/6 LD50, the inhibitory effect of yungumycin (15 mg.kg-1) on sarcoma 180 was similar to that of 5-FU (40 mg.kg 1), showing 72% and 70% tumor inhibition, respectively. Initially, gougerotin was reported as an antibacterial antibiotic without mentioning its antitumor activity. The present studies demonstrate that yungumycin (gougerotin), by oral administration, may be useful in cancer chemotherapy. PMID- 9206266 TI - [Inhibitory effects of protein kinase C inhibitors on tumor necrosis factor induced bovine pulmonary artery endothelial cell injuries]. AB - The effects of protein kinase C(PKC) inhibitors 1-(5-isoquino-linylsulfonyl)-2 methylpeperazine (H-7) and quercetin on tumor necrosis factor (TNF) were studied in cultured bovine pulmonary artery endothelial cells (BPAEC) in vitro. Incubation of BPAEC with TNF caused a significant increase in percent lactate dehydrogenase (LDH) release, stimulation of EC-dependent neutrophils (PMN) adhesion to BPAEC and inhibition of BPAEC DNA synthesis and proliferation. All these were restored by both H-7 and quercetin. The IC50 of H-7 and quercetin was 9.7 and 10.8 mumol.L-1 for the inhibition of LDH% release; 19.5 and 16.7 mumol.L 1 for the inhibition of TNF-induced PMN-EC adhesion; 7.0 and 6.1 mumol.L-1 for TNF-induced inhibition of DNA synthesis and 8.7 and 11.36 mumol.L-1 for proliferation. These results suggest that PKC inhibitors H-7 and quercetin protect BPAEC from TNF induced injuries and PKC play an important role in EC activation by TNF. PMID- 9206267 TI - [A double column double pH solid phase extraction and capillary GC/FID method for rapid simultaneous determination of acidic and basic drugs in human plasma]. AB - A capillary gas chromatographic method for the rapid simultaneous identification and quantitation of acidic and basic drugs in human plasma is presented. A special double column solid phase extraction device was designed, in which two X 5 resin columns can extract drugs at different pH. The detection limits for acidic and basic drugs can reach 0.5 microgram.ml-1, while the time needed is only half of that when using commercial solid phase extraction cartridges. PMID- 9206268 TI - [Preparation of diclofenac sodium controlled release pellets and multiple-dose evaluation in healthy volunteers]. AB - The yield of pellets obtained in preparing diclofenac sodium (DC-Na) controlled release pellets was affected by the lactose/starch ratio of the feed granules, viscosity of the binding solution and residence time in the rolling pot. The pellet shape was found to be correlated with surface tension of the binding solution. The concentration and composition of the coating solution and pellet size distribution were responsible for the agglutination in coating. The controlled release pellets (CRP) and commercial tablets (CT) of DC-Na were administrated orally to healthy volunteers by multiple-dose. The steady-state condition, based on trough plasma concentration on day 3-5, was achieved on day 3. A great difference in plasma drug concentration fluctuation index (FI) between CRP (0.476 +/- 0.0484) and CT (0.935 +/- 0.092) was observed (P < 0.05) during steady-state. The area under the plasma drug concentration-time curve for a 0-12 h interval (AUC) on day 5 of CRP (6.493 +/- 0.4169 micrograms.h.ml-1) and CT (7.551 +/- 0.4745 micrograms.h.ml-1) was shown to be not significantly different (P > 0.05). The relative bioavailability in the human was about 86%. PMID- 9206269 TI - Enhancing effect of essential oils on the penetration of 5-fluorouracil through rat skin. AB - Three essential oils as penetration enhancers for 5-fluorouracil (5-FU) were studied using excised rat skin. The oils used were eucalyptus, peppermint and turpentine. Azone was used for comparison. The enhancing effect of the oils was found to be less than that of azone, but all the oils used enhanced the permeation of 5-FU. Eucalyptus oil was found to be the most active, causing about 60 fold increase, while peppermint and turpentine caused 46 and 28 fold increase, respectively. Eucalyptus oil was further studied by grading it into 5 fractions according to difference in boiling points. It was found that their activities increased as their boiling point increased. With all enhancers increased partition coefficients were observed but the diffusion coefficient values obtained were comparatively higher. The mode of action of these accelerants may be described by combined processes of partition and diffusion, the diffusion process being dominant. PMID- 9206270 TI - [Progress in the studies on medicinal plants of the genus Zanthoxylum Linn]. PMID- 9206271 TI - [Cloning and sequencing the isopenicillin N synthetase(IPNS) gene from Streptomyces cattleya]. AB - Great homology existed between IPNS genes from surphur-containing beta-lactam antibiotics producers including procaryotes and eucaryotes. A DNA homologous band was confirmed in S. cattleya by Southern blot analysis using IPNS gene from S. lipmanii as a probe. A recombinant plasmid containing the cyclase gene involved in thienamycin biosynthesis and IPNS gene was obtained by complementary cloning with mutant from S. cattleya. DNA sequencing revealed that the IPNS gene of S. cattleya consists of 963 bp encoding a protein of 321 amino acids with ATG as start codon, TGA as stop codon. Pairwise comparison of the predicted amino acid sequences showed 56% and 64% similarity with IPNSs of S. clavuligerus and S. lipmanii, respectively. PMID- 9206272 TI - [Study on the test and evaluation of the toxicity of the strains preparation of the different subspecies of Bacillus thuringiensis and on the research into its standardization]. AB - In this paper using the preparation of Bacillus thuringiensis of USA standard sample-subsp. kurstaki HD-1-S-1980 (H3a3b) the toxicity to different subspecies preparations such as subsp. dendrolimus U strain(H4a4b), subsp. galleriae C88 strain (H5a5b) is tested and evaluated. And product's standardization is also researched. Comparing their toxicity to various test-insects between the standard sample prepared from U strain and USA, standard sample, the boilogical determined results are as follows: the toxicity of U strain standard sample is respectively 18666.6 IU/mg(test-insects: Dendrolimus punctatus, 2nd larva), 22956.5 IU/mg(test insects: Plutella kylostella, 2nd larva). The above-mentioned toxicity evaluations are higher than that of 16000 IU/mg of USA standard sample. The comparison in susceptibility shows as follows: suscepticibility of plutella kylostella is stronger than that of Dendrolimus punctatus. The products from U strain and C88 strain are trial-produced in medium-scale. The product's toxicity to test-insect Dendrolimus punctatus is determined using USA standard sample. The results are obtained as follows the toxicity evaluation of U strain products is 36444.4 IU/mg; that of C88 strain products is 28521.7 IU/mg. The both are obviously higher than that of USA standard sample. This proves that different subspecies strain's toxicity to the same test-insect is with defference and it is an important way to raise product's toxicity by improving and optimizing process in production. The whole experiment shows it is feasible to rectify the evaluation of the toxicity of our country's present products using USA standard sample, Dendrolimus punctatus and Plutella kylostella as test-insets. And it is simple and convienient as well as save time and save effort to make statistical analysis in computer with Basic program. To do statistical and operational analysis using our designed basic programming by micro-computer is more accurate and repid than by caculator. PMID- 9206273 TI - [Purification and characterization of S-layer protein from Aeromonas hydrophila]. AB - The regular surface protein array (S-layer) present on Aeromonas hydrophila J-1 was composed of a single species of protein of apparent molecular weight 51500. This protein was extracted from whole cells by treatment with 0.2 mol/L glycine hydrocholoide (pH4.0). The protein was purified by Sephadex G-200 gel filtration chromatography and an ion exchange chromatography on DEAE-cellulose. Amino acid composition analysis showed that the protein contained 36.8% hydrophobic amino acids. But the protein did not confer hydrophobicity to the cell surface when present as an intact S-layer by salt aggregation test. ELISA and immunoblotting with two different polyclonal antisera against surface exposed-(PM) and non surface-exposed epitopes (PR) of the protein revealed that the sensitivity of PM was higher than that of PR. Antigenic diversity of the S-layer proteins from 20 bacterial samples was analysed by ELISA with PM and PF1 (polyclonal antiserum against Aeromonas hydrophila TF7 S-layer protein). The S-layer proteins were distinguishable from the extracellular toxin of the homogeneous strains in antigenic and biochemical characterization and the S-layer proteins were one of the main protective antigens. PMID- 9206274 TI - [Study on the death of Escherichia coli induced by hydrostatic pressures]. AB - The effect of hydrostatic pressure on the death of E. coli was studied in this paper. The results indicated that E. coli could be killed by hydrostatic pressure above 800 bar. At 2300 bar E. coli was totally killed in 30 minutes. The time course of E. coli death induced by pressure indicated that the most E. coli was killed in the first 10 minutes after the pressure was applied. It was also found that the lower temperature favored killing E. coli under pressure. PMID- 9206275 TI - Practising oncology via telemedicine. AB - Although there are increasing numbers of telemedicine programmes in the USA, few have offered teleoncology services, so that the role of telemedicine in the practice of clinical oncology has yet to be fully defined. Telemedicine has been used successfully for direct patient care in Kansas. It is also a method of providing supportive care for the cancer patient, including assessments of pain and nutrition. In addition, televised tumour conferences and nursing education courses can help smaller communities develop a level of expertise that allows patients to be treated locally. Telemedicine may well be used in future for access to national and international cancer experts, and for participation in new cancer treatment protocols through cooperative group trials. When practising oncology via telemedicine, there are unique problems, including issues regarding technology (interactive video and radiograph review) and practice (patient/oncologist preferences and doctor-patient communication). Very little has been published in the area of tele-oncology so far, and studies concerning its efficacy, cost-effectiveness and the best organizational structure are still in progress. However, telemedicine appears to be a useful technique in the practice of oncology. PMID- 9206276 TI - Urban teleradiology in Hong Kong. AB - We investigated four major teleradiology programmes in Hong Kong. We analysed the overall organizational background of each programme, the context of its implementation, the choice of technology and the decision-making process, and the subsequent dissemination of the technology. Our review suggests that the success of a telemedicine programme is contingent not only on good technology but also on effective management of issues pertaining to human and organizational factors. At the departmental level, the context of the implementation of telemedicine is crucial. At the institutional level, success depends on planning and management. At the professional level, continued education, periodic meetings and seminars are all effective means of promoting telemedicine among health-care professionals and administrators. At the national level, the Hong Kong experience suggests that telemedicine can be effectively developed by starting with urban-based programmes. PMID- 9206277 TI - Performance tests of a satellite-based asymmetric communication network for the 'hyper hospital'. AB - The Hyper Hospital is a prototype networked telemedicine system which uses virtual reality. We measured the performance of a novel multimedia network based on satellite communications. The network was a hybrid system consisting of a satellite channel in one direction and a terrestrial channel in the other. Each user was equipped with a standard satellite communication receiver and a telephone connection. Requests from the users were sent by modern and telephone line and responses were received by satellite. The user requests were initiated by clicking buttons on a World Wide Web browser screen. The transmission rates of satellite and normal telephone-line communications were compared for standardized text data. Satellite communication was three to five times faster. The transmission rate was also measured for standardized graphical data (GIF format). With a file size of about 400 kByte, satellite-mediated communication was 10 times faster than telephone lines. The effect of simultaneous access on performance was also explored. For simultaneous access of nine users to a single graphics file, 78% of the transmission speed was obtained in comparison with that of a single user. The satellite-based system showed excellent high-speed communication performance, particularly for multimedia data. PMID- 9206278 TI - Effect of camera performance on diagnostic accuracy: preliminary results from the Northern Ireland arms of the UK Multicentre Teledermatology Trial. AB - The diagnostic accuracy of realtime teledermatology was measured using two different video cameras. One camera was a relatively low-cost, single-chip device (camera 1), while the other was a more expensive, three-chip camera (camera 2). The diagnosis obtained via the videolink was compared with the diagnosis made in person. Sixty-five new patients referred to a dermatology clinic were examined using camera 1 followed by a standard face-to-face consultation on the same day. A further 65 patients were examined using camera 2 and the same procedure implemented. Seventy-six per cent of conditions were correctly diagnosed by telemedicine using camera 2 compared with 62% using camera 1. A working differential diagnosis was obtained in 12% of cases using camera 2 compared with 14% using camera 1. The percentage of 'no diagnosis', wrong and missed diagnoses was halved using camera 2 compared with camera 1. These results suggest that the performance of the more expensive camera was superior for realtime teledermatology. PMID- 9206279 TI - Distributed clinical neurophysiology. AB - We have developed a consultation forum for clinical neurophysiology in Finland. The system connects local digital electroencephalography (EEG) recording and analysing networks using a high-speed asynchronous transfer mode (ATM) network. Clinicians can obtain a second opinion using interactive data and video consultations or using data-only consultations. In addition, the system can be used for off-line review of pre-recorded data. During a one-month evaluation, 66 EEG recordings were made altogether in Satakunta Central Hospital and consultations were required on 12 occasions. Nine of them were data-only consultations and three were data and video consultations. A data consultation lasted 15-20 min and a data and video consultation 35-45 min. Clinically, there were numerous benefits for the hospitals. The system established a link to a centre of excellence for second opinions or continuing education. It also helped with on-duty arrangements and enabled the construction of national data banks. PMID- 9206280 TI - A portable unit for remote monitoring of pacemaker patients. AB - The number of people with pacemakers has been steadily increasing over the past 20 years. At present there are about 2,500,000 in the world. All these patients must have a checkup twice a year in hospital, which involves the allocation of significant resources by the national health systems, not to speak of the inconvenience it entails for elderly subjects. Telemedicine can provide a partial solution to these problems. The present work describes a prototype system that transmits the patient's electrocardiograms and the main characteristics of the pacemaker to a monitoring centre. This remote monitoring system will be beneficial to patients and health-care organizations in terms of quality of life for the former, and costs and efficiency for the latter. PMID- 9206281 TI - Image transfer and computer-supported cooperative diagnosis. AB - The KAMEDIN system was designed as a low-cost communication tool as part of a computer-supported cooperative work project that included synchronized user interaction, telepointing and audioconferencing. During a five-month field trial, it was used for medical image transfer and cooperative diagnosis in 14 clinics and medical departments in Germany. During the field test, 297 teleconsultations were performed via ISDN and 875 MByte of data were transferred. An image compression ratio of 2-3 was obtained, so that the total quantity of data transferred corresponded to 14,000-21,000 magnetic resonance images or 3500-5250 computerized tomography images. Furthermore, 694 local sessions were conducted for the preparation of teleconsultations and the review of transferred images. Participants learned to handle the KAMEDIN system in a few hours. This was mainly owing to the design of the user-oriented graphical user interface and the restriction of the system to a set of essential image-processing functions. PMID- 9206282 TI - Benefits and pitfalls of telemedicine in neurosurgery. PMID- 9206283 TI - International telemedicine. PMID- 9206284 TI - Current questions about teleradiology. PMID- 9206285 TI - A limited free eye test? PMID- 9206286 TI - An underage teenager who requests contraception. PMID- 9206287 TI - Strategies for managing watering eyes. PMID- 9206288 TI - Red eye: avoid the pitfalls. PMID- 9206289 TI - Highlights of the year in ophthalmology. PMID- 9206290 TI - What's new in laser treatment? PMID- 9206291 TI - Who should be screened for glaucoma? PMID- 9206292 TI - Diagnosis and treatment of dry eyes. PMID- 9206293 TI - A GP guide to the new asthma guidelines. PMID- 9206294 TI - Chickenpox. PMID- 9206295 TI - Poverty: initiatives to protect children from its effects. PMID- 9206296 TI - MRGCP: what to expect from the second oral. PMID- 9206297 TI - The demise of clinical skills? PMID- 9206298 TI - [External ear abnormalities: syndromic and genetic aspects]. AB - Morphological defects of the external ear represent a significant class of congenital abnormalities because of their overall frequency and their impact on affected people. All types of congenital defects are involved in the study of the ear. The external ear is an important site for the study of minor phenotypic variations. Associated abnormalities are frequent and lead to syndromes identification. Some genes involved in syndromes with abnormal external ear as a feature have been recently cloned by molecular genetics. PMID- 9206299 TI - [Reconstruction of the pinna in cases of microtia]. AB - Pessimism about the results of surgical reconstruction of the pinna in cases of microtia is unjustified. Currently there are two techniques available which give excellent results if conditions are favourable: these are the techniques of Brent and of Nagata. This article describes both of these techniques, whose common principle is the use of autologous costal cartilage sculptured to give the contour of a normal ear, and the areas where they are different. The author has carried out 154 reconstructions using the Brent technique and 92 using the Nagata technique, and gives a critical analysis of the results obtained by these two methods. PMID- 9206300 TI - [Techniques of total reconstruction of the pinna]. AB - Reconstruction of the pinna must be adapted for each individual case. Surgical techniques which use autologous costal cartilage are the most reliable. During the 1970s Burt Brent described a technique in 4 stages which gives very good results. More recently, Satoru Nagata has proposed total reconstruction of the pinna in two stages. For cases where there is insufficient spare skin, a tissue expansion prosthesis may be used in cases with extensive scarring and fibrosis, the superficial temporalis fascia flap is useful. Bone-anchored prostheses are reserved for those cases in which surgery is contra-indicated for general reasons. Based on our experience of 119 reconstructions of the pinna, we discuss the various techniques which may be used. PMID- 9206301 TI - [Functional surgery of major aplasia of the ear. Current status]. AB - One of the challenges of functional surgery for microtia is to prevent the postoperative otorrhea. The subgaleal flap or the galeal flap, which are the two components of the superficial temporalis flap as it is currently used in plastic surgery, are, each of them, convenient for covering the ear canal and give a good support for the skin grafting. The dry ear canal can fit an air conduction hearing aid. PMID- 9206302 TI - [Reconstruction and ear prosthesis: what are the indications?]. AB - There are two procedures available for treatment of the agenetic pinna: reconstructive plastic surgery using costal cartilage, and bone-anchored prostheses. Both techniques are described, and the indications clarified. It is important to emphasise that all of the possible alternatives must be within the repertoire of the surgeon, so that the treatment can be adapted to suit the individual case. PMID- 9206303 TI - [Submental island flaps. Surgical technique and possible variations in facial reconstruction]. AB - The submental flap is an island myocutaneous flap supplied by the submental artery divided from the facial artery. Large skin paddles (up to 7 x 18 cm.) can be raised, perfectly matching with facial color. As an island flap it allows coverage of the homolateral oral cavity or the homolateral face (except the medial forehead region). Including the facial vessels in its pedicle allows a safe free transfer. A greater are of rotation is obtained transferring the flap on its distal pedicle. One can include in the flap a segment of internal basilar margin. The resulting scar is perfectly hidden under the mandible. The operative protocol is now well defined and makes the flap easy to use. 21 flaps have been transferred, resulting in a single partial necrosis of a flap extremity. PMID- 9206304 TI - [Superficial pediculated temporal fascia flap in the reconstruction of the oropharynx]. AB - The authors emphasize the interest of using the fascial temporal superficial flap for the reconstruction of an oropharynx after tumoral removal. The intrinsic qualities of this aponevrotic flap (rich vascularization, plasticity, finness, absence of after effects for the doner) have made it a particularly innovative instrument in this field. They expose their experience through nine patients and specify the place of this mean of reconstruction. PMID- 9206305 TI - [Infrahyoid musculocutaneous flaps: anatomical bases and indications in cervicofacial oncologic surgery]. AB - The infrahyoid myocutaneous flap has been described by Wang in 1986. Its use seems still limited, although its obvious interest in head and neck reconstructive surgery, especially in oropharyngeal area. Authors expound anatomic bases of this flap, and a 62 case's experience confirming its potential. PMID- 9206306 TI - [Inferior blepharoplasty: the transconjunctival approach]. AB - Described in 1924 by J. Bourguet, the transconjunctival approach seems to have experienced a resurgence of interest in recent North American publications. The incision, made under local anaesthesia along the inferior border of the tarsal cartilage, provides a plane of dissection which can be either preseptal, in which the approach to the bags is made through the septum as in the classical approach, or alternatively retroseptal, a route which respects the integrity of the septo orbicular suspension, and allows a direct approach to th bags. Removal of fat should be limited to the excess tissue which protrudes when pressure is applied to the globe, using the orbital rim as a landmark. After haemostasis, closure is provided by means of a gathering stitch which is removed on the 4th day. The indications are young patients who present with bags with no excess of either skin or muscle (preseptal approach), dark-skinned patients in whom there is a risk of scarring (keloid), and older patients (retroseptal approach) who present with hyperlaxity of the septo-ligamentous mechanism, and in whom the "round eye" or an ectropion might result from a classical approach. In this last group, the excessive skin may be excised a minima without undermining. Finally, in cases of fat remnants after the classical approach, the transconjunctival approach offers an easy solution. Complications are extremely rare. It is for this reason that the transconjunctival approach is tending to become the standard approach for many modern authors. PMID- 9206307 TI - [Frontal and subcutaneous temporal closed lifting without endoscopy: technique and indications]. AB - This new technique of fronto-temporal facelift first described by Tranck Trepsat, while permitting very limited incisions for an endoscopic approach, also allows results at least as good without use of the endoscope. It also provides a simple and elegant solution to the problem of temporary fixation in all closed facelifts: the temporary self-tapping self-breaking rivet, this paper is based on the first series of 28 patients. A distinction is drawn between two frontal regions, identified by their characteristic mode of ageing, and treated selectively by "periscopy". For the fronto-lateral region and the temple: here it is the cutaneous layer which ages and sags; it is treated by subcutaneous dissection. For the median frontal region; it is the whole cutaneo-muscular tissue which ages and sags. This is treated by subgaleal elevation with a rugine from the three superior marking points. This new technique of facelift is based on a more detailed analysis of the fronto-temporal ageing process, and on selective treatment for it. It achieves a return to normal appearances. The main factor is lateral lifting. The short scars and the avoidance of the need for video-endoscopy make this a quick and economical procedure. PMID- 9206308 TI - [Video-assisted endoscopic lifting: development]. AB - Now the Endoscope is used differently in video-assisted Face-lifts. Authors describe their evolution for 2 years and a half. In the beginning. It was used to correct glabellar frowns. Then, the endoscope served to elevate the forehead by a subperiosteal approach. Sometime it was combined with a subperiosteal malar dissection in an Endoscopic full face-lift. Bad results on the eyebrows and eyelids complications involved the authors to modify their strategy: By changing the dynamic of muscles: lateral section of the orbicularis occuli muscle arises the frontal muscle to definitely lift up the eyebrow: this is called the "myotomy box"; by preferring a subgaleal, retro-orbicularis dissection, under the malar fat pad and the platysma muscle in a deep plane but always above the zygomaticus major muscle, protector of the facial nerve in the buccal space; by developing new endodissectors efficient and less aggressive to spread the deep planes without damage; by an acute control of the fronto-temporal and cervico-facial deep dissection using new endoscopic valves; by treating precise damaged areas with Limited Endoscopic Face-Lifts in a same harmonious deep dissection instead of large multiplanar underminnings. PMID- 9206309 TI - [Reconstruction of lip isocele]. AB - The surgical treatment of labial carcinoma must allow the complete excision of the tumor. This must be performed by using an immediate reconstruction procedure with respect of the lip function and with a satisfying aesthetic result. To achieve this goal, we choose the isosceles lip reconstruction, according with Meyer's advancement flap technique, described in 1965. By using this method we can restore up to the 2/3 of the lower or upper lip and also the lip commissure, after malignant tumor surgery. For total lip reconstruction, we have to combine this method with an Abbe-Estlander flap. Our paper presents this original lip reconstruction technique and analyses the results obtained. We especially base our conclusions on functional tests, but also on aesthetical considerations. We underline the interest of this choice of surgical procedure, permitting its adaptation to all cases of lip reconstruction, with immediate possibility of reconstruction, usually in one stage, without compromising either the function or the aesthetic aspect of the reconstructed lip. PMID- 9206310 TI - [Protruding ears]. PMID- 9206311 TI - Does justice require less precision than chemistry? PMID- 9206312 TI - Bayesian framework for the evaluation of fibre transfer evidence. AB - The application of the likelihood ratio, derived in a Bayesian framework, to different case scenarios involving fibre evidence has permitted the authors to identify and evaluate the dominant parameters and their effect on the likelihood ratio. Moreover, it has been emphasized that these parameters are not only defined by the conditions of the contact, but also by the strategy chosen by the defence. PMID- 9206313 TI - A study on the random distribution of a red acrylic target fibre. AB - This project was a target fibre study carried out by the European Fibres Group. Thirty-eight participating laboratories in nineteen countries searched four hundred and thirty five randomly selected top outer garments. The acrylonitrile/vinyl acetate co-polymer target fibre matched those in a red acrylic scarf, marketed in nine of those countries. The sales figures for the last five years were between 5- and 10,000 scarves annually. Matching fibres were found on only two garments, one fibre per garment. The possible origin of these two fibres is discussed. The result showed that the chances of finding a 'collective' of target fibres on a piece of clothing selected at random can be said to be extremely remote. PMID- 9206314 TI - A preliminary study of the analysis of Cannabis by supercritical fluid chromatography with atmospheric pressure chemical ionisation mass spectroscopic detection. AB - A rapid method is described for the analysis of Cannabis products by supercritical fluid chromatography (SFC) coupled to atmospheric pressure chemical ionization-mass spectroscopic (APCI-MS) detection. The method had a shorter analysis time than GC-MS methods, without the need for derivatization prior to analysis. It was also faster than HPLC methods, with better resolution and definitive identification. Linearity of detector response to cannabidiol, delta 8 tetrahydrocannabinol, delta 9-tetrahydrocannabinol and cannabinol was established, the detection limits for mass on column being 0.55 ng, 1.20 ng, 0.69 ng and 2.10 ng respectively. The technique offers a means by which Cannabis products can be definitively identified in a single chromatographic run. Application to casework samples is described. PMID- 9206315 TI - The collection of data from findings in cases of sexual assault and the significance of spermatozoa on vaginal, anal and oral swabs. AB - The construction of a database to hold information obtained from the laboratory analysis of items in cases of sexual assault is described. Examples of the usefulness of the database are described. For vaginal, anal and oral swabs, the persistence of spermatozoa in relation to time since intercourse is presented and discussed. PMID- 9206316 TI - An improved method for the preparation of combs for use in hair combing kits. AB - Hair combing kits are used to collect samples of fallen hair, fibres and other trace evidence from the hair of the suspect and injured party. The time taken to prepare the hair combing kits has been significantly reduced by using the comb loader described in this paper. PMID- 9206317 TI - Identification and quantification of amphetamine and analogues by capillary zone electrophoresis. AB - A free zone capillary electrophoresis (CZE) system, with diode array detection, was used in the analysis of ephedrine and 9 amphetamines. The method was quantitative, offering better resolution than gas chromatography (GC), without the need for sample derivatisation. Under hydrodynamic injection, the concentrations of drug at which detection limits were reached lay between 13 and 68 micrograms/ml, with detection at 214 nm, these limits being of the same order of magnitude as those of GC. Analysis of casework samples by the two methods demonstrated the advantages of CZE with diode array detection over conventional GC for amphetamine identification and quantification. PMID- 9206318 TI - The acquisition of breaking and broken glass. AB - A set of experiments tested the idea that an examiner can distinguish glass acquired by backward fragmentation from that acquired by contact with already broken glass. A window was broken. The exterior surface of the breaker's clothing was brushed and hair-combings taken and these were searched for glass fragments. Wearing the same clothing the breaker then cleared up the broken glass and further hair-combings and brushings of the clothing surface were taken. The size, shape and presence or absence of an original flat surface was recorded for the recovered fragments. The distributions within these parameters were compared for the two methods of acquisition and association between the method and the parameters was tested using contingency tables. It was found that fragment shape does not identify the method of acquisition and that a high proportion of fragments retaining the original outer surface shows that the glass was acquired by backward fragmentation. PMID- 9206319 TI - CUSUM: a credible method for the determination of authorship? AB - Authorship attributions based upon the CUSUM (Cumulative Sum Analysis) method are still being presented in court proceedings as evidence despite the findings of several independent researchers that the method is unreliable. This paper describes the shortcomings of the method, presents further examples of its failures and summarises the findings of other researchers. The CUSUM method is seen to be discredited and should not be accepted as providing reliable evidence of authorship of either the written or the spoken word. PMID- 9206320 TI - Linear synthetic aperture modes for ultrasonic pulse-echo imaging. AB - A unified approach is developed to characterize and compare the most widely used two-dimensional ultrasound pulse-echo imaging procedures. The methods of monostatic, bistatic and multistatic time-domain holography are theoretically investigated with regard to line spread functions, resolution capabilities, and the effects of multiple scattering and object inhomogeneities. Analytical derivations and computer simulations show the superiority of monostatic holography in applications such as nondestructive testing, where in most cases the assumptions generally made in the calculation of the inverse scattering formulae are justified. However, if higher order scattering and/or the spatial variations of the sound speed play a significant role, then multistatic imaging is more advantageous. A modification of this latter mode, with fixed focus on transmission, is commonly used in medical imaging B-scan systems, but implementation of the full algorithm would offer improved resolving power and reduced side lobe levels throughout the image plane. PMID- 9206321 TI - Learning a new bimanual coordination pattern: reciprocal influences of intrinsic and to-be-learned patterns. AB - According to dynamic pattern theory, intrinsically stable bimanual coordination patterns affect, and are affected by, the acquisition of a new coordination pattern. In Experiment 1, subjects practiced either a 45 degrees or a 135 degrees relative phase pattern for 4 days; in Experiment 2, they practiced a 90 degrees relative phase pattern over 6 days. Retention tests were conducted 4 weeks after the last practice session in both experiments. Performance on both the in-phase (0 degree) and anti-phase (180 degrees) patterns was also measured on each day. Contrary to predictions, the experiments revealed that reciprocal effects between the intrinsic patterns and the new pattern were only temporary, and did not affect learning in any permanent way. As well, learning a new pattern was not differentially affected by its relation to an intrinsic pattern. PMID- 9206322 TI - Covert visual orienting across the lifespan. AB - Covert visual orienting was examined over a span of human life ranging from six to 73 years. The observer's task was the speeded discrimination of "X" from "O1" but of primary interest was the effect of a location cue that appeared prior to the target. Both an abrupt stimulus cue and a voluntary information cue were studied using response time (RT) measures. Eye movements were monitored to control for differences in the ability to maintain fixation. Experiment 1 showed that there were very few age differences in stimulus-cued orienting. In contrast, there were important differences when orienting was intentional. In comparison with young adults, children were less able to sustain orienting over time, and senior adults required more time to use the cue. Experiment 2 tested the relation between stimulus and information cues when they both occurred prior to a given target. All age groups were able to use information cues in the presence of conflicting stimulus cues, but young adults were better able to do so than either children or senior adults. These results are interpreted as support for the view that separate mechanisms underlie stimulus-based versus information-based spatial orienting. PMID- 9206323 TI - Depressive symptoms and alterations in sucrose taste perception: cognitive bias or a true change in sensitivity? AB - Previous studies have reported elevated taste thresholds in depressed subjects, but those studies did not control for changes in response bias. The current study used signal detection analyses to address this shortcoming. Sucrose detection thresholds were measured (1) in subjects with high and low Hamilton Depression Rating Scale (HAM-D) scores who did not meet standard criteria for current Major Depressive Episode (MDE); and (2) in subjects who did fulfil standard criteria for MDE. Subjects with low HAM-D scores produced significantly more false alarms than the other two groups, but taste sensitivity, as indexed by d', did not vary significantly across groups. These results suggest that changes in response bias underlie previously reported increases in sucrose taste thresholds in depressed subjects. PMID- 9206324 TI - Reading-based interference in cognitive arithmetic. AB - Do number-fact retrieval (4.6 =?) and numeral reading (e.g., transcoding 46 into "forty six") access the same retrieval structures? Data from the present experiment suggest that they do. Under instructions to respond quickly, adults performed simple multiplication problems oriented horizontally or vertically with operands presented simultaneously or with a 500 ms preview of one operand. Errors involving congruent operand intrusions (e.g., 2.8 = 24 or 9.6 = 36) were most frequent when conditions afforded the left-to-right encoding sequence that is standard for reading multi-digit numbers. Ninety percent of such intrusions involved the correct answer to another simple multiplication problem (e.g., 4.8 = 28) rather than a miscellaneous answer (e.g., 4.8 = 38). This high rate of arithmetically related intrusions suggests that numeral reading and verbal production of number facts involve a shared representational system. PMID- 9206325 TI - Dendritic cells: a novel cellular component of the rat incisor enamel organ appearing in the late stages of enamel maturation. AB - Immunocompetent cells in the enamel organ of rat incisors were examined immunohistochemically using OX6, ED1, and ED2 monoclonal antibodies known to recognize the Class II MHC molecules, a monocyte-macrophage lineage, and residential macrophages, respectively. The OX6 immunopositive cells (MHC cells) were located exclusively in the enamel maturation zone. MHC cells increased in number in the incisal direction and occasionally extended cytoplasmic processes deep into the ameloblast layer. Migration of MHC cells in the ameloblast layer were also encountered. MHC cells lacked phagolysosomes and could be distinguished from typical macrophages. ED2 immunopositive cells were not seen in the enamel organ. ED1 positive cells displayed identical localization to MHC cells except that some appeared in the transitional zone. MHC cells could not be seen in the enamel organ of rat molar tooth germs. Our data confirmed the presence of a large population of "dendritic" immunocompetent cells in the enamel organ of rat incisors and characterized the ultrastructural features of these cells. Biological significance of the immunocompetent cells in the enamel organ during amelogenesis needs to be clarified. PMID- 9206326 TI - Immunohistochemical localization of signal transduction pathways during amelogenesis: an initial exploration. AB - This study was undertaken to map signal transduction pathway (STP) components uniquely associated with the four major receptor groups and their related STPs in association with the events involved in amelogenesis in the rat. Whole-head, freeze-dried sagittal sections were obtained at the level of the maxillary first molars and picked up on transparent adhesive tape. The sections were not decalcified or fixed, providing optimum conditions for immunohistochemical (IHC) localization. Antibodies to pathway components Gs alpha, Gi alpha, Gq alpha, Sos 1, Grb-2, p125Fak, Jak2, and Vav were localized. The respective patterns of localization indicate that the Gq alpha-linked, the receptor tyrosine kinase initiated, and the integrin receptor-initiated pathways are involved in the proliferating pre-ameloblast cells. In the differentiating and differentiated ameloblasts, the Gs alpha-linked cAMP pathway is involved, apparently reading a factor(s) released by the dentin matrix. The Gq alpha-linked, the receptor tyrosine kinase-initiated, the integrin receptor-initiated, and the cytokine receptor-initiated pathways are also up-regulated in the proximal ends of the ameloblasts. These observations indicate that all four of the major receptor groups are involved in amelogenesis and that the role of classes of ligands not previously implicated in enamel formation must now be considered. It seems that the cells of the enamel organ respond to the appearance and disappearance of autocrine and paracrine growth factors, but they also up-regulate specific STPs to enable them to respond to circulating hormones and growth factors whose concentrations in the extracellular fluids remain relatively constant. PMID- 9206327 TI - Primary structure of the porcine 89-kDa enamelin. AB - The primary structure of the 89-kDa enamelin found in porcine secretory enamel at an early stage of development was investigated. The fragments of the enamelin cDNA were amplified by polymerase chain-reaction from the first-strand enamelin cDNA, and were sequenced. The results indicated that the 89-kDa enamelin consisted of 627 amino acid residues and had a molecular mass of 70,448. A hydrophobic domain is located in the region of the 21st-62nd amino acid residues of the molecule. Acidic domains are located in two regions of the molecule-one in the region of the 135th-238th amino acid residues and the other in the C-terminal region. A basic domain is located in the region of the 239th-360th amino acid residues. The results also indicated that the low-molecular-weight enamelins were fragments derived from a prototype enamelin. PMID- 9206328 TI - Albumin gene expression during mouse odontogenesis. AB - Albumin protein is present in developing teeth of several species. Oligomer primers and cRNA probes specific for albumin were designed to perform RT-PCR, and for in situ hybridization, respectively. In situ hybridization failed to reveal albumin expression in any tooth cells, however, albumin PCR products were amplified from tissues adhering to the roots of developing teeth from four-week old mice. It is concluded that this source is not the primary source of albumin protein found in developing enamel, because of the location and level of expression of albumin mRNA in periodontal tissue. PMID- 9206329 TI - Radioautographic study of the incorporation of (3H)-choline into the phospholipids of secretory ameloblasts and enamel of normal and essential-fatty acid-deficient rats. AB - (3H)-choline, a precursor for phosphatidylcholine (PC) and sphingomyelin (SM), was injected into rats killed after 4, 24, 48, and 96 hrs. Radioautography carried out on malachite-green/aldehyde-fixed tissues demonstrated that labeled choline was incorporated into cells and further released into the extracellular matrix. In predentin, labeling decreased rapidly, whereas in dentin, silver grains formed a stable band. In contrast, labeling was still high at 48 and 96 hrs in secretory ameloblasts as well as in the forming enamel. This indicates that ameloblasts are actively involved in the synthesis of membranes. Membrane remnants of the ameloblasts could be released into the forming enamel. In rats fed with an essential fatty-acid-deficient (EFAD) diet for 42 days, (3H)-choline uptake was delayed and reduced in pulp cells and odontoblasts, and consequently the migration of labeled phospholipids into dentin. The influence of the EFAD diet on secretory ameloblasts was limited. No difference was detected between normally fed and EFAD-fed rats in the forming enamel. PMID- 9206330 TI - Transient accumulation of proteins at interrod and rod enamel growth sites. AB - Conceptually, there should be a brief interval in time when newly secreted proteins "pile up" at secretory sites just outside the membrane of ameloblasts. Indeed, previous cytochemical studies have suggested that glycosylated and/or sulfated glycoproteins accumulate at enamel growth sites. Colloidal gold lectin cytochemistry and immunocytochemistry with antibodies to enamel proteins and phosphoserine, combined with cycloheximide and brefeldin A to inhibit protein synthesis and secretion, were applied to characterize the distribution of newly formed proteins at enamel interrod and rod growth sites. Although enamel growth sites show a "rarefied" appearance, the results indicate that one or more subclasses of enamel proteins accumulate near the cell surface at sites where elongation of enamel crystallites contributes to thickening of the enamel layer. These proteins are glycosylated and/or phosphorylated and, at least in the case of the glycosylated ones, are rapidly processed after they are released extracellularly. In contrast, immunolabeling for amelogenins is generally weaker near the cell surface and more intense at a short distance away from the site where crystallites elongate. The data suggest that the enamel proteins accumulating at growth sites likely belong to the non-amelogenin category and play a transient role in promoting the lengthening of crystallites. It is concluded that areas near the ameloblast membrane where certain enamel proteins accumulate in fact constitute the equivalent of a mineralization front. PMID- 9206331 TI - Comparative HPLC, SDS-PAGE, and immunoblot analyses of dental enamel proteins. AB - The primary structures of amelogenins expressed from different genes vary because of DNA sequence divergence and variations in alternative RNA splicing. The pattern of splicing is unique for each amelogenin gene yet investigated, even when two copies of the gene are expressed in the same cell. Despite the high conservation of amelogenin sequences, diversity in the pattern of RNA splicing leads to significant differences in the number and character of amelogenin isoforms in the developing enamel matrix. Since conservation of molecular structure is an indicator of functional significance, we compared enamel protein preparations from rat, porcine, rabbit, and opossum developing tooth organs. Enamel extracts were fractionated by reversed-phase high-performance liquid chromatography (HPLC) and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Western blot analyses were performed with polyclonal antibodies raised against recombinant murine amelogenin and the polypeptide encoded by murine exon 4. The opossum enamel extract produced the simplest chromatogram, suggesting that fewer proteins are secreted into the developing enamel matrix. The predominant opossum amelogenin has an apparent molecular mass of 28 kDa and reacts strongly with the recombinant amelogenin antibody but is not recognized by the murine exon 4 antibody. Opossum amelogenin mRNA was amplified with murine amelogenin primers specific for the amino- and carboxyl-terminal coding regions. The mobility of the amplification products on 4% agarose gels indicates that the leucine-rich amelogenin polypeptide (LRAP) is expressed in the opossum and that the major amelogenin is larger than its homologue in the mouse. We conclude that the alternative splicing of amelogenins pre-dates the metatherian and eutherian divergence over 100 million years ago. PMID- 9206332 TI - Developmental changes in the pH of enamel fluid and its effects on matrix resident proteinases. AB - The objectives of this study were to measure pH in developing enamel at progressively older (more mature) stages of amelogenesis in vivo, and then to formulate synthetic enamel fluid mixtures that approximated these pH values for in vitro studies. The ultimate goal was to characterize the molecular weights of proteinases visualized by enzymograms incubated in synthetic enamel fluid using gelatin and casein as substrates. For most experiments, the proteinases were extracted en masse from small freeze-dried enamel strips directly into a non reducing sample preparation buffer. In some experiments, we pre-treated the enamel strips with acetic acid to determine if this common method for demineralization and protein extraction caused any changes in the activity levels of the enamel proteinases. In other experiments, we first soaked enamel strips in synthetic enamel fluid to determine solubility of the proteinases within an aqueous phase. The results indicated that the pH of developing enamel remained fairly constant near pH 7.23 across the secretory stage, but it was generally more acidic (6.93) and fluctuated in focal areas between mildly acidic (6.2-6.8) and near-neutral (7.2) conditions across the maturation stage. The pH then slowly rose to near 7.35 when the enamel was almost mature (hard). The acidic conditions were generally inhibitory to most enamel proteinases, but there were some caseinase activities in mid-maturation-stage enamel near 23-30 kDa which appeared to be activated by weakly acidic conditions (pH 6.28). Pre-treatment of enamel samples with 0.5 M acetic acid markedly altered the overall profile of enamel proteinases, causing activation of some latent proteinase activities and permanent inhibition of other activities. Most proteinases in whole homogenates were insoluble in synthetic enamel fluid. This suggests that they may be tightly bound, directly or indirectly, to matrix proteins or mineral components in situ. PMID- 9206333 TI - Possible actions of metalloproteinases found in porcine enamel in an early secretory stage. AB - In an outermost layer of porcine secretory enamel, metalloproteinases were detected by enzymography with gelatin used as a substrate. When the sample extracted from the outermost layer of the secretory enamel was incubated with calcium ions at 37 degrees C prior to electrophoresis, an increase of the 34-kDa proteinase activity and a decrease of the 76- and/or 78-kDa proteinase activities were observed. The results suggest that the metalloproteinases mediate the conversion from 76- and/or 78-kDa proteinases to the 34-kDa proteinase or the activation of a latent type of the 34-kDa proteinase, and that their activities are regulated by free Ca ions. PMID- 9206334 TI - Crystal growth in dental enamel: the role of amelogenins and albumin. AB - Amelogenin-mineral interactions were investigated using an in vitro binding approach. Rat incisor enamel matrix proteins (mainly amelogenins) were dissolved in synthetic enamel fluid and allowed to equilibrate with deproteinised developing enamel crystals. The results showed that amlogenin proteins of 21, 23, 24, 26 and 27-kDa (corresponding to nascent and partially degraded amelogenins) were associated with the crystals whilst the lower Mr amelogenins (< 21 KDa) remained free in the synthetic enamel fluid. These data suggest the nascent and partially degraded amelogenins may interact with developing enamel crystals and could influence their growth. Albumin-mineral interactions were investigated by extracting developing rat incisor enamel with synthetic enamel fluid. Insoluble material (including the enamel crystals) was then further extracted with 0.1 M phosphate buffer (pH 7.4) to desorb any mineral bound proteins. Western blotting using anti-albumin antibodies showed that almost all of the albumin from the secretory stage enamel and a significant proportion of the albumin present in early transition stage was extractable in the synthetic enamel fluid. However, synthetic enamel fluid did not extract albumin from late transition or maturation stage tissue, which could only be removed following further extraction with phosphate buffer. Albumin degradation was apparent during the transition and maturation stages, where it is degraded and ultimately removed. This binding pattern may be related to amelogenin degradation and removal during the transition stage, permitting albumin access to the previously obscured crystal surfaces. That the secretory stage matrix appears to "protect" secretory stage crystals from albumin may be an important consideration in the aetiology of enamel hypoplasias (i.e. incomplete crystal growth) and when using dissociative extraction procedures for the identification of mineral bound proteins. PMID- 9206335 TI - The rat amelogenin gene--some aspects of evolution and expression. AB - This study presents data to support the hypothesis that a major portion of the coding sequence of the amelogenin gene may have arisen by tandem duplication of internal sequences which as a consequence has introduced several additional potential RNA splice acceptor sites into the sequence. This duplication of splice sites has led to an increase in the heterogeneity of amelogenin forms found in developing enamel. By screening a rat enamel organ cDNA library for alternatively spliced products, it appears that as much as 20% of the amelogenin mRNA molecules may be alternatively spliced forms. PMID- 9206336 TI - High expression of human amelogenin in E. coli. AB - A human cDNA, encoding for the 175-amino-acid human amelogenin, was prepared by RT PCR from tooth bud mRNA and sub-cloned into pGEX-KG expression plasmid for over-expression in E. coli. The expressed protein was characterized by SDS-PAGE Western blotting, and N-terminal amino acid sequencing. PMID- 9206337 TI - Expression patterns of RNAs for amelin and amelogenin in developing rat molars and incisors. AB - We have recently identified a novel RNA sequence in ameloblasts, coding for amelin (Cerny et al., 1996). In the present paper, its expression has been compared with that of amelogenin in developing incisors and molars of rats, by means of in situ hybridization of paraffin sections. The RNAs for both amelin and amelogenin were highly expressed in secretory ameloblasts. The expression of RNA for amelogenin gradually decreased in the post-secretory ameloblasts. In contrast, the RNA expression for amelin remained high in post-secretory ameloblasts up to the stage of fusion between dental and oral epithelia at the time of tooth eruption. We suggest that amelin might be involved in the mineralization of enamel or in the attachment of ameloblasts to the enamel surface. The whole-mount in situ hybridization procedure is described for the first time in dental research. It proved to be a useful method and confirmed the results of the conventional in situ hybridization. PMID- 9206338 TI - Gene expression and localization of amelogenin in the rat incisor. AB - Gene expression and localization of amelogenin were studied in the developing rat incisor by the methods of in situ hybridization and immunohistochemistry. ISH revealed the first expression of amelogenin mRNA in the inner enamel epithelium of the cervical loop. The signals were clearly observed in pre-ameloblasts in the region bordering on predentin formation and became more intense toward the cells on the initial enamel matrix secretion. The maximal signals were found in the cytoplasm of secretory ameloblasts. From the terminal secretion zone, the signals then became gradually weaker toward the incisal edge but were still evident in the cytoplasm of shortening, transitional ameloblasts and those at the early maturation stage. No signals were found in the cells of the stratum intermedium and stellate reticulum throughout amelogenesis. Immunohistochemistry by means of an antibody against amelogenin C-telopeptide consisting of 12 amino acids revealed immunoreaction in the secretory ameloblasts reacting to the ISH. When a polyclonal antibody against amelogenin was used, immunoreaction was found in the distal ends of ruffle-ended ameloblasts (RA) in the maturation zone. Those results indicated that amelogenin is synthesized by ameloblastic cells from the inner enamel epithelium to the early maturation stage and is then resorbed by the RA. PMID- 9206339 TI - Toward understanding the function of amelogenin using transgenic mice. AB - The purpose of this study was to establish transgenic mouse lines as a tool to investigate the function of amelogenin during mineralization by causing ectopic production of amelogenin and studying its effect. The mouse amelogenin (mAme) was cloned from a 16-day-old whole mouse embryo cDNA library and was determined to be "full-length" mouse amelogenin (with a complete coding region) by comparison with the mouse amelogenin reported previously by Snead et al. (1985) and Lau et al. (1992). The overexpression construct contained: (1) the rat osteocalcin (OC) promoter (1.8 kb); (2) the adenovirus splicing casettes, including introgenic (Int) sequence (0.3 kb); (3) the full-length mAme cDNA (0.8 kb); and (4) the polyadenylation signal sequence from the pSG5 mammalian expression vector. Both Southern blotting and polymerase chain-reaction (PCR) analyses were performed, by means of a specific probe and a pair of oligodeoxynucleotides to OcIntmAme(A)+, respectively. The animals which showed transgene-positive in both analyses were further used to establish F1 animals. Heterozygocity was confirmed with F1 animals by PCR analysis of DNA from the F0 x FVB/N pups. Three independent transgenic F1 heterozygous lines (640t, 706t, and 708t) have now been established. The generation of F2 homozygous lines is under way. The heterozygous transgenic animals are currently being analyzed for alterations in the morphology and structure of various bone tissues. PMID- 9206340 TI - Disturbed enamel mineralization in a rat incisor model. AB - Possession of full-thickness hard enamel appears to be one of the indispensable life-saving characteristics of rats. Previous studies by Suga and his colleagues and by others demonstrated that various types of malformation are evoked in continuously erupting rat incisors. In the current report, we directed our effort to oversee various types of enamel malformation caused experimentally in rat incisors. We surveyed the specimens collected by Suga and his colleagues, as well as specimens we obtained. From the results, it is conceivable that perturbation of the programmed sequential events during enamel development is a major factor in the establishment of enamel malformation. Animal studies with either 1 hydroxyethylidene-1,1-bisphosphonate (HEBP) or a multidentate phosphonic acid (EDTPO) confirmed that dentin mineralization provides a certain inductive effect on the secretion of enamel matrix and subsequent enamel crystallization. Our recent studies using anti-microtubular agents led to the conclusion that the acceleration of mineralization in outer enamel is a type of enamel malformation, most likely due to disruption of the cellular regulation of calcium transport under severe toxic regimens. In future work, experimental approaches combining measurements of kinetic factors with static observation of enamel lesions are required before we can gain a comprehensive understanding of the pathogenesis of disturbed enamel mineralization. The kinetic factors to be considered include the rates of tissue apposition and tooth eruption which determine the total volume of tooth substance formed, and the rate of mineral accretion. Furthermore, information as to the composition, crystallinity, solubility, and mechanical properties of enamel defects is needed before we can assess the susceptibility of teeth having those lesions to caries and other physico-chemical attacks in the oral environment. PMID- 9206341 TI - Magnesium and carbonate in enamel and synthetic apatites. AB - This study aimed to: determine the Mg and CO3 distribution in the outer (surface), middle, and inner (closest to the enamel-dentin junction, EDJ) layers of human enamel; and determine the factors affecting the incorporation of Mg into synthetic apatites and the consequence of such incorporation on the properties of the apatites. Results demonstrated that the concentrations of Mg, CO3, and organic components increased from the surface to the inner layers close to the EDJ and a difference in crystallinity from the outer to the inner layers. Initial results indicated that the extent of dissolution of the inner layer enamel is greater than that in the outer or surface enamel. Results on synthetic apatites showed the following: (1) Limited Mg incorporation into apatite was dependent on solution [Mg/Ca] molar ratio, temperature, pH, and the presence of CO3 or fluoride (F); (2) incorporation of Mg causes reduction in crystallinity and an increase in the extent of dissolution of the apatite; (3) the negative effect of Mg on the properties of apatites is synergistic to that of CO3 and antagonistic to that of F; and (4) exposure to acid of Mg-containing apatites causes the dissolution of Mg-rich apatite and precipitation of Mg-poor apatite. The observed decrease in the [Mg/Ca] of enamel and synthetic apatites after acid exposure may explain the observed 'preferential loss' of Mg and CO3 in the initial stages of caries. PMID- 9206342 TI - Lattice fringe continuity in the absence of crystal continuity in enamel. AB - Since high-resolution transmission electron microscopy (HRTEM) provides information on a nearly atomic level, the confidence level with this method is very high. Thus, when lattice fringe continuity is found between two enamel crystals in proximity, such continuity has been taken as evidence of crystal fusion (Daculsi and Kerebel, 1977). Similarly, selected-area dark-field (SADF) electron microscopic imaging has been used to study the axial and spatial orientation of crystals. These studies have shown that there is apparent continuity between enamel and dentin crystals (Arsenault and Robinson, 1989). This observation supported the hypothesis that enamel crystallites are initiated by crystallites in dentin. We have used both HRTEM and SADF methods to identify instances of spatial relationship between crystallites in sections of rat incisor enamel and shark enameloid. In each instance of apparent continuity, goniometric tilting was used to examine the continuous interface. All instances where two crystallites seemed to come into contact, and where HRTEM imaging showed the lattice fringes to be directly continuous, were separated into individual crystallites when the specimen was tilted a few degrees. Thus, adjacent crystallites can show lattice fringe continuity in the absence of real crystallite contact. When instances of overlapping crystallites were examined by SADF imaging, the overlapping crystallites gave a single bright image. Goniometric tilting revealed separate crystallites. Thus, neither lattice fringe continuity nor image continuity under SADF can be used as evidence of crystal continuity unless goniometric rotation and tilting are applied when spatial relationships are suspected. PMID- 9206343 TI - Anion translocation through the enamel organ. AB - The objective of this study was to determine whether cells of the secretory- and maturation-stage enamel organ of rats contain anion translocation mechanisms similar to those found in other ion-regulating epithelia. Sodium bromide (Br) was used to localize the distribution of anions in the enamel organ. Furosemide, an inhibitor of the Na-K-2Cl co-transporter and other anion transporters, was administered with NaBr or sodium fluoride (F) to investigate if halogens other than Cl can use these transport mechanisms. We obtained the data by using freeze fracture and freeze-drying methodology in conjunction with scanning and transmission electron microscopy (SEM, TEM) and energy-dispersive x-ray spectroscopy (EDS). The secretory- and maturation-stage enamel organ prevented Br from entering the enamel matrix. Br was localized in the Tomes' processes, but not in the enamel matrix, strongly suggesting that the distal intercellular junctions of ameloblasts are "tight". Furosemide disrupted anion transport to allow not only Cl but also Br to enter the forming enamel matrix. Periodic administration of high F doses promoted the formation of bands of disrupted enamel, reflecting the periodicity of F administration. The same concentration of F administered with furosemide increased the severity of disrupted enamel, resulting in "blisters" and pits in the maturing enamel. The enamel "blisters" contained pools of small, disorganized enamel crystallites. The group receiving furosemide only displayed normal enamel structure but had increased Cl in the enamel matrix. This study provides evidence that anion transporters, possibly the Na-K-2Cl co-transporter, function to regulate anion translocation, including F, to the enamel matrix in secretory- and maturation-stage enamel organ. These mechanisms may explain why the ionic composition on the cellular side of the anion barrier is different from that of the enamel matrix. PMID- 9206344 TI - Quantitative immunocytochemistry of Ca(2+)-Mg2+ ATPase in ameloblasts associated with enamel secretion and maturation in the rat incisor. AB - Our previous studies revealed intense membrane-associated labeling for Ca(2+) Mg2+ ATPase (Ca(2+)-pump) in secretory and maturation ameloblasts in the rat incisor, both by enzyme cytochemistry and by immunohistochemical techniques. The purpose of the present study was to map the distribution of Ca(2+)-pump protein at the cellular and subcellular levels by means of a Ca(2+)-pump-specific monoclonal antibody and electron microscopic immunogold cytochemistry. Tissue specimens were dissected from secretory, early, and late enamel maturation zones. We quantified results by comparing gold particle densities over ameloblast lateral and distal plasma membrane regions, supranuclear cytoplasm, regions of the ruffled borders, and nuclei. The highest concentration of gold particles was seen over the distal membranes of early-maturation ameloblasts relative to those in late-maturation and secretory stages. Cytoplasmic labeling was less than that of the distal and lateral membranes, and gold particles located over nuclei were considered to be due to non-specific binding. These results are consistent with our earlier findings and suggest a role for the plasma membrane Ca(2+)-pump in the regulation of calcium availability to mineralizing enamel. PMID- 9206345 TI - Properties of heterogeneous apatites containing magnesium, fluoride, and carbonate. AB - Biological apatites present in the mineral phases of normal and pathological calcifications contain magnesium, Mg, and carbonate, CO3. As a consequence of fluctuations in the composition of the micro-environment, these apatites may sometimes form by heterogeneous precipitation. The purpose of this study was to investigate the properties of (Mg, CO3)-apatites formed heterogeneously in the presence of fluoride, F. Two types of fluoridated (Mg, CO3)-apatites formed from solutions with low and high levels of Mg were prepared at 80 degrees C, pH 7.4. We prepared FMgCO3-MgCO3AP (Type 1) by adding the F-containing solution to those containing calcium, Mg, and phosphate ions during the first half of the precipitation period. We prepared MgCO3-FMgCO3Ap (Type 2) by adding the F containing solution during the final half of the period. The apatites were analyzed by x-ray diffraction (XRD), infrared absorption spectroscopy, and scanning electron microscopy (SEM). SEM and XRD analyses showed evidence of mixed crystals in the heterogeneous apatites. The presence of Mg inhibits, while F promotes, apatite crystal growth. In addition, Mg incorporation increased with increasing fluoride concentration. The extent of dissolution in acid buffer of both types of heterogeneous apatites increased with Mg: Type 1 > Type 2. These results suggest that the crystal and dissolution properties of heterogeneous fluoridated (Mg, CO3)-apatites are greatly affected by the mode of F incorporation and Mg concentrations in the environment. PMID- 9206346 TI - Effects of accelerated eruption on the enamel of the rat lower incisor. AB - The effect of accelerated eruption of the rat lower incisor on enamel was studied in a series of segments obtained when the incisor was cut repeatedly out of occlusion over a five-week period. The segments were ground, cleaned, acid etched, observed with SEM, and analyzed with EDX. Pigmentation was lost within 11 days. Pigmented superficial enamel was more acid-resistant than the rest of the enamel, but this quality decreased with decreasing iron content. Hypomineralized enamel first appeared in a restricted area at the mesio-labial angle of the tooth in the 6th-7th segment obtained after 11-14 days. Later, hypomineralization became more generalized. All enamel zones were retained throughout the experiment. The geometry of the prism pattern was affected. The angle between prism rows and the enamel-dentin junction increased from 44 degrees to 48 degrees, while the angle of decussation increased from 60 degrees to 70 degrees. The angle between the enamel surface and prisms in the outer enamel was more difficult to assess, but tended to increase from about 25 degrees to 29 degrees. However, the prisms retained their incisal direction. The connection between enamel and dentin was partly disrupted from about the 9th segment onward. The depth of the mesial concavity of the enamel-dentin junction decreased from about the 10th segment onward. Accelerated eruption affects all stages of enamel formation and is a suitable and predictable model for studying regulatory mechanisms in amelogenesis. PMID- 9206347 TI - Enamel mineral composition of normal and cystic fibrosis transgenic mice. AB - The ability of ameloblasts and the enamel organ to control the influx of ions into the developing enamel is of considerable interest. The development of transgenic mice lacking a cAMP-regulated chloride channel, the cystic fibrosis transmembrane conductance regulator (CFTR), provides a model that may prove valuable for the study of ion regulation in developing teeth. The purpose of this investigation was to characterize the mineral content of normal and CF mice. Five homozygous and five heterozygous adult mice having the CFTR knockout transgene were evaluated. The mice were killed with CO2 and their mandibular incisors removed, embedded in methacrylate, and sectioned, and enamel particles from the incisal region were then dissected for analysis. Each particle was analyzed for its calcium, phosphorus, and magnesium content. The normal mice had a mean mineral content of 80.5%, in contrast to the CF mice, that had markedly hypomineralized enamel (mean = 51.5%). The calcium/phosphorus ratios were similar for both groups of mice and were compatible with the enamel consisting primarily of hydroxyapatite mineral. The enamel magnesium content was significantly elevated in the CF mice (mean = 3560 ppm) compared with the normal mice (mean = 2280 ppm). Normal mouse enamel was highly mineralized, while the CF mouse enamel mineral content was significantly reduced and had an elevated level of magnesium. The altered mineral content of CF mouse enamel indicates that CFTR could play an important role in ion regulation and consequently mineralization of mouse enamel. PMID- 9206348 TI - Pulpal temperature changes during low-power hard-tissue CO2 laser procedures. AB - Thermal insult to pulpal tissue is recognized as a major limitation to the use of lasers for dental hard-tissue procedures. This study examined thermal changes at the level of the dental pulp in human molar teeth irradiated with a CO2 dental laser using a pulsed mode of operation. Sectioned molar teeth were exposed, in vitro, to CO2 laser radiation. The laser parameters were those used clinically for laser desensitization and laser-enhanced fluoride treatment. Fissure regions and root surfaces were irradiated. For settings which might reasonably be used clinically, the temperature rise was not of a magnitude which would be expected to cause pulpal inflammation or necrosis. With regard to thermal properties of tooth structure, times taken to reach the maximum temperature reduced, and times taken to cool to baseline increased with increasing laser exposures. PMID- 9206349 TI - Early tensile bond strength between dentin and composite resin mediated by bonding agents. AB - The aim of this study was to evaluate, in vitro, the early tensile bond strength values of three dentin bonding agents (Scotchbond, Tenure and Gluma) to dentin. Fifteen non-carious, extracted, human third molars were used in this study. The bond strengths were calculated in an Instron test machine and expressed in MN/m2. The results were evaluated statistically using the Student-test. The in vitro tensile bond strengths of the three bonding agents to dentin showed higher values than the tensile bond strength of composite resin alone and differences among the bond strengths were significant (P < 0.05). Greater bond strength values were obtained with the Gluma bonding system than with Scotchbond or Tenure systems. PMID- 9206350 TI - SEM analysis of smear layer removal after manual and automated handpiece root canal preparation. AB - Scanning electron microscopy was used to analyze the smear layer removal after root canal preparation by a manual technique and by an automated handpiece, the Canal Finder System (CFS). When 1% sodium hypochlorite was used as the irrigating solution, both manual and CFS techniques showed root canal walls with a dense smear layer obscuring the dentinal tubules entrance plus a large amount of debris. Root canal walls of the group of teeth treated with a chelating agent (EDTA) for 5 min and a final flush, after the preparation, with 1% sodium hypochlorite as an irrigating solution showed the cervical, middle and apical thirds extremely smooth and clear. PMID- 9206351 TI - Effect of CO2 laser on Class V cavities of human molar teeth under a scanning electron microscope. AB - The purpose of this study was to evaluate the effects of CO2 laser on dentin of class V cavities of extracted human molar teeth using a scanning electron microscope. SEM showed a smooth area with concentric lines formed by melting with subsequent recrystallization of dentin, areas of granulation, vitrified surface, numerous cracks, and irregular areas of descamative dentin. These data indicate that CO2 laser (4 and 6 watts) produces dentin alterations and limit its clinical applications. PMID- 9206352 TI - External and internal anatomy of mandibular molars. AB - The external and internal anatomy of 628 extracted, mandibular first and second molars was studied. The external anatomy was studied by measuring each tooth and by observing the direction of the root curvatures from the facial surface. The internal anatomy of the pulp cavity was studied by a method of making the teeth translucent. PMID- 9206353 TI - Chemical analysis of the liberation of calcium and hydroxyl ions from calcium hydroxide pastes in connective tissue in the dog--Part I. AB - The objective of this research is to chemically analyze calcium hydroxide pastes added to three hydrosoluble vehicles having different acid-base characteristics using polyethylene tubes implanted in subcutaneous connective tissue in a dog, evaluating the liberation of calcium and hydroxyl ions over a period of 7, 30, 45 and 60 days. The three vehicles were saline, anesthetic, and polyethylene glycol 400. Chemical analysis of the liberated calcium ions was done by means of conductimetry using EDTA for titration. Liberation of hydroxyl ions was determined by analogy of calcium ions liberated, which are in direct proportion to the molecular weight of calcium hydroxide. PMID- 9206354 TI - Dentinogenesis imperfecta type II: case report. AB - A case of dentinogenesis imperfecta type II is described. The authors also present a brief literature review and focus on the difficulty in treating such cases. PMID- 9206355 TI - Association of a temporomandibular disorder and Eagle's syndrome: case report. AB - We report a clinical case of Eagle's syndrome which required dental intervention due to the presence of exacerbated symptoms indicating an association with a temporomandibular disorder. The therapeutic dental procedures used were an occlusal splint and temporary removable partial dentures. Surgical removal of the styloid process on the left side was later performed as a medical option. PMID- 9206356 TI - Periodontal disease progression in type II non-insulin-dependent diabetes mellitus patients (NIDDM). Part I--Probing pocket depth and clinical attachment. AB - Periodontal disease progression of 30 type II diabetic patients (NIDDM) and 30 patients in whom diabetes was not detected was evaluated. Age ranged from 30 to 77 years. To determine the periodontal condition, probing pocket depth and periodontal attachment loss were measured; to determine the metabolic control of the patients, glycosylated hemoglobin and fasting glucose were measured. At the end of the study, the diabetic group was divided into three subgroups, according to the metabolic state of the patients: controlled patients, moderately controlled patients, and poorly controlled patients. Comparing the diabetic and the control groups as a whole, there was no statistically significant difference in probing pocket depth, but significance (P < 0.01) was observed for attachment loss. When diabetic patients were divided into subgroups, significant differences were observed between the poorly controlled and the control groups (P < 0.01) for both the probing pocket depth and periodontal attachment. The glycosylated hemoglobin test was more reliable than the fasting glucose analysis. PMID- 9206357 TI - In vitro action of various carbamide peroxide gel bleaching agents on the microhardness of human enamel. AB - The authors verified a decrease in human enamel microhardness after application of the following carbamide peroxide gel bleaching agents for 8 hours daily for 1 week: 10% Nite White, 16% Nite White, Opalescence, Karisma Alpha and Perfect Smile. Statistical analysis showed that these agents caused a decrease in enamel microhardness. Nite White (16%) was the most effective in reducing enamel microhardness and Opalescente the least effective. Nite White (10%), Karisma Alpha and Perfect Smile fell into a statistically intermediate position. PMID- 9206359 TI - Diameter of axons in the lingual nerve of the mouse. A preliminary investigation. AB - The lingual nerves from ten mice were examined so that normal axonal populations could be determined. After perfusion fixation, they were removed and processed, and sections were taken from nerves for transmission electron microscopy. The fiber-diameter spectrum of unmyelinated fibers for the mouse lingual nerve is characterized by a unimodal curve with the more pronounced peak in the medium diameter fiber range (0.28 micron). The spectrum from the myelinated fibers also shows a population of axons of the lingual nerve (0.22-3.2 microns) that reflects different functional specializations. These data establish some baseline values for morphological evaluation of the effects of experimental lingual nerve damage. PMID- 9206358 TI - Comparison of a new test for the measurement of resting whole saliva with the draining and the swab techniques. AB - A quantification method for measuring whole saliva is described. This whole saliva test (WST) consists of a Whatman paper strip, is easily carried out, innocuous, low-cost and single use. Due to its characteristics, it could be considered as the oral equivalent of Schirmer's tear test. A sample of 159 healthy subjects (81 males and 78 females; mean age 31.62 years) participated in this comparative study of this new procedure and two other tests, the draining and the swab test. Correlation was statistically positive among the three types of tests. PMID- 9206360 TI - Study of the correlation between the gingival immunologic defense index and parameters associated with dental caries. AB - The objective of the present study was to determine the possible existence of a correlation between the gingival immunologic defense index (GIDI) and parameters associated with dental caries such as number of teeth with cavities, number of colony forming units (CFU) of streptococci of the mutans group, decay missing filling deciduous teeth (dmft), and decay missing filling permanent teeth (DMFT). Since no correlation was detected with the above parameters, we conclude that the simple presence of caries or of CFU is not sufficient to stimulate IgA production and/or secretion. For this to occur, an unfavorable action of these parameters on gingival health is needed, with the production of gingival inflammation. PMID- 9206361 TI - Oral health and periodontal status in Brazilian elderly. AB - A total of 104 elderly persons between the ages of 60 and 89 were examined at the "Geraldo de Paula Sousa" Health Center, Sao Paulo, State of Sao Paulo. The state of their oral health was very poor, insofar as 4.29 (71.5%) of the sextants were shown to be null, while 0.12 and 0.13 sextants showed deep periodontal pockets > or = 6 mm in the ages from 60-70 and more than 70 years of age, respectively. The level of knowledge about periodontal disease and dental plaque was very deficient; only about 52% of the population under study reported having visited a dentist in the last two years. We conclude that greater odontological attention is needed for the elderly age group, as there are no large-scale community service centers in Brazil for this population group. PMID- 9206362 TI - Surgical management of premalignant lesions of the oral cavity with the CO2 laser. AB - The management of patients with premalignant and malignant lesions of the oral cavity can present problems. The potentially invasive nature of premalignant lesions together with their large extent influences the treatment. The common modalities of treatment of these lesions are surgical excision, cryotherapy, electrosurgery and radiotherapy. Recently, CO2 laser surgery has become available. Less pain, little bleeding, minimal post-operative edema, reduced risk of infection, and low recurrence rates were advantages observed following CO2 laser surgery in the mouth when compared to other modalities of treatment. Healing following CO2 laser surgery progressed well with little postoperative scarring and re-epithelialization was complete after 4-6 weeks. The newly formed epithelium appeared normal and was soft on palpation. PMID- 9206363 TI - Comparative study of the direct-lift and platinum foil techniques in the marginal discrepancy of collarless metal ceramic restorations. AB - This study was carried out to evaluate the marginal discrepancy of collarless metal ceramic restorations, using a combination of three different techniques to manufacture the porcelain butt margin with two brands of body porcelain. Statistical analysis showed no significant difference between the techniques or brands of body porcelain used in this study. PMID- 9206364 TI - Natal teeth in cleft lip and palate patients: a scanning electron microscopy study. AB - Natal and neonatal teeth may occur in patients up to 30-days after birth. The presence of two neonatal teeth in cleft lip and palate patients is reported in this study describing the structural aspects of the enamel and the dentin, using scanning electron microscopy. PMID- 9206365 TI - Is antibiotic prophylaxis required for endodontic treatment? AB - The stable prevalence of infective endocarditis since the advent of antibiotic prophylaxis for patients at-risk reflects the increasing polymicrobial etiology of such infections not associated with dental procedures. In addition to concerns for the growing crisis for antibiotic-resistant bacteria, the need for controlled clinical trials to determine the continued efficacy of prophylactic regimens for endodontic and other dental procedures cannot be overstated. PMID- 9206366 TI - Endodontic treatment of teeth with apical periodontitis using calcium hydroxide: a long-term study. AB - The aim of this study was to evaluate clinically and radiographically the long term results of endodontic therapy. A total of 172 mature teeth with periapical radiolucencies with and without symptoms were treated endodontically using calcium hydroxide paste as the intracanal medicament and a calcium hydroxide containing root canal sealer. In 58 teeth, the dressing was accidentally or intentionally extruded into the lesions. All cases were followed up for a period of 2-5 years. The teeth in which the dressing was extruded did not show a different healing pattern from the ones treated conventionally. The complete healing rate for all cases was 80.8% while incomplete healing had taken place in 7.6% of the cases. PMID- 9206367 TI - Area-metric analysis of dye leakage for evaluation of sealing ability of root canal obturation techniques. AB - Root canal sealing ability of obturation techniques has been assessed in vitro with various methods. The majority of the methods employ microleakage tracers and particularly dyes. In vitro measurements of dye penetration are either linear or volumetric. Area-metric analysis is a three dimensional registration method of dye leakage. The purpose of the present study was to evaluate in cleared teeth area-metric analysis as opposed to linear analysis of dye penetration. Forty freshly extracted single rooted human teeth were used. Instrumentation was carried out using Hedstroem files with a step back technique. The root canals were obturated using Roth sealer and qutta-percha cones and lateral condensation technique. The roots were then subjected to dye leakage tests under vacuum of 50 mmHg for 20 min. India ink was used as the tracer. The roots were randomly divided in two groups. In Group A the roots were cleared and linear measurements of the dye penetration were recorded The roots in Group B were ground stepwise transversally and subjected to area metric analysis. The results showed that area metric analysis enabled sufficient recordings of the dye leakage patterns and the volume of the dye penetration could also be calculated. PMID- 9206369 TI - Compatibility between standardized endodontic finger spreaders and accessory gutta-percha cones. AB - Twenty samples of each size designation of finger spreaders and accessory gutta percha cones were randomly selected from a proprietary standardized system currently used for lateral condensation procedures. Their diameters were measured with a calliper and their morphological variations at the areas of their apical thirds were analyzed with the scanning electron microscope. Results of the measurements indicated that there were high degrees of variation between all spreaders and their corresponding accessory gutta-percha cones (p < 0.001). SEM examination showed the presence of many morphologic irregularities and lack of uniformity between spreaders and cones of the same size. Our results emphasize the need for more precise criteria to fulfill the requirements of standardization by the manufacturer to provide an accurate method for root canal obturation. PMID- 9206368 TI - Push-out strength and SEM evaluation of resin composite bonded to internal cervical dentin. AB - The bond strength of a resin composite used with a dual-cured dentin bonding system to internal cervical bovine dentin was evaluated using a direct or indirect placement technique. Teeth were sectioned transversely to produce 4 mm thick specimens. The root canals were enlarged to a standardized taper, treated with a dentin bonding system, and filled with a light-cured resin composite using either direct, incremental composite placement or indirect composite placement of a pre-polymerized composite inlay. The debond stress of indirectly placed restorations using a composite inlay was 8.5 (SD +/- 2.7) MPa which was significantly greater (p < 0.0001) than the value of 5.0 (SD +/- 1.9) MPa for composite placed in a conventional, incremental manner. SEM evaluation revealed the indirect placement technique demonstrated increased resin tag density and length as compared to the direct technique. Enhanced retention of resin composite to endodontically prepared dentin treated with a dentin bonding system was obtained by using a composite inlay technique as opposed to direct, incremental buildup of the material. PMID- 9206370 TI - Analysis of the surface cut by sonic files. AB - The purpose of this investigation was to compare 3D profilometry and SEM analysis of a polished surface of bovine bone instrumented by sonic files. Two situations where investigated, i) no operator assisted movement, ii) operator assisted movement. Heliosonic, Rispisonic and Shaper files were investigated at full power setting and an interfacial load of 100 grams. The specimens were subjected to analysis using a 3D Form Talysurf prior to being sputter coated with gold and viewed under an SEM. The 3D surface analysis was found to be complimentary to SEM evaluation in that each provided additional information to the other. 3D analysis proved to be excellent for showing the topographical nature of the cut surface and gave a better indication of depth than the SEM. Debris was however more apparent on SEM evaluation. Each file showed distinctive characteristics in the shape that was cut. The cutting appeared to be as a result of abrasion with no operator assisted movement and was as a result of the longitudinal file action. However when movement was superimposed on this action continuous chip formation was apparent especially for the Rispisonic and Shaper files. PMID- 9206371 TI - Endodontic retreatment: a rational approach to non-surgical root canal therapy of immature teeth. AB - The specificity of retreating immature teeth involves generally a broadly wide open apex and/or an inverted root canal conicity (apical opening wider than the root canal orifice), a periapical pathology leading often to a continuous canal exudation and thin and fragile dentinal walls. Nonsurgical endodontic retreatment of immature teeth consists at first in removing coronal and root canal iatrogenic obstructions in order to regain total canal patency. Apexification is then the treatment of choice, preceding a final gutta-percha obturation. The present article outlines a step-by-step rational approach to the root canal retreatment of immature teeth, pertaining to their biomechanical and pathological specificity. PMID- 9206372 TI - Root perforations: classification and treatment choices based on prognostic factors. AB - Root perforations are common complications of endodontic treatment or post preparation and often lead to tooth extraction. Successful treatment depends mainly on immediate sealing of the perforation and prevention of infection. Several factors affect the achievement of these goals, most important of which are: time of occurrence, size, and location of the perforation. A classification of root perforations, based on the above factors, is presented to assist the clinician in the choice of the treatment protocol which will give the best possible results when a perforation is diagnosed. PMID- 9206373 TI - Therapeutic delivery of calcitonin to inhibit external inflammatory root resorption. I. Diffusion kinetics of calcitonin through the dental root. AB - Insertion of calcitonin into root canals of monkey teeth has been shown to inhibit external inflammatory root resorption and suppress inflammation. Regulation of this therapeutic event depends upon the rate of arrival (diffusion) of the hormone at sites of resorptive activity. In the present study, the diffusion characteristics of calcitonin through the dental root in an extracted human-tooth model are described, and the role of cementum in the diffusion process is also addressed. Root-canals were endodontically prepared to form a reservoir for [125I]-calcitonin, and macerated to remove organic material from dentinal tubules. In teeth with intact cementum, an initial period of delay (4-5 h) prior to the detection of calcitonin at the external tooth-root surface was followed by a rapid release of the calcitonin during the first 10.5 h (rate peaks at 6 h). Slower, sustained releases of calcitonin through intact cementum were measured for the following 9 days. Removal of cementum, to expose "smear-free" dentine, resulted in an earlier efflux of calcitonin (2 h) at external tooth surfaces and increased amounts of calcitonin release over 9 days. Biphasic delivery of calcitonin by such internal diffusion mechanisms suggests that loss of cementum will enhance therapeutic availability, while prolonged delivery to intact external dental-root surfaces following early intra-canal placement may also be useful for the therapeutic prevention of external inflammatory root resorption. PMID- 9206374 TI - Therapeutic delivery of calcitonin to inhibit external inflammatory root resorption. II. Influence of calcitonin binding to root mineral. AB - Experimentally-induced external inflammatory tooth-root resorption can be inhibited by therapeutic doses of calcitonin. Such doses can be delivered by an intrinsically slow diffusion pathway, from a reservoir in endodontically-debrided root canals, via the dentinal tubules. While the kinetics of this journey have been followed in an earlier report, the binding characteristics of calcitonin to the tooth mineral, which will be responsible, in part, for these kinetics, have not been reported before. The current study examines the binding potential of calcitonin to root mineral and addresses the potential role of non-specific binding proteins. A modified Scatchard plot indicated that a simple non-reactive type of ligand binding exists between calcitonin and root mineral, represented by a small number of identical binding sites. This interaction is both strong and reversible. Furthermore, it appears to be time-dependent with more time being required for the residual ligands to interact with the diminishing numbers of free calcitonin-binding sites. While preloaded [125I]-calcitonin could be incompletely (75-91%) displaced from dental-root material by non-radioactive calcitonin, its release was slow over 23 h. Calcitonin was four times as effective as bovine-serum albumin in competing for common "calcitonin binding sites" on macerated dental-root material. Thus, even in the presence of extraneous protein, calcitonin will bind tightly but reversibly to tooth-root material, making it a good candidate for therapeutically protracted delivery to external root surfaces from root canals. PMID- 9206375 TI - An in vitro study of the efficacy of mouthguard protection for dentoalveolar injuries in deciduous and mixed dentitions. AB - Sports-related dental trauma remains a risk for children and adolescents. Although mouthguards provide protection, up to 25% of dentoalveolar injuries can occur with a mouthguard in place. This study examined the effect of mouthguard protection in an in vitro model. A total of 97 sheep mandibular segments with incisors at four developmental stages (early deciduous, ED, n = 37; middle deciduous, MD, n = 20; late deciduous, LD, n = 18; mixed dentition, PD, n = 22) was used. Customised pressure formed mouthguards (MG) provided protection from trauma produced by a servohydraulic materials testing machine to test incisors. Injuries were examined clinically, radiographically and by dissection. Mean forces required to produce dentoalveolar injury were significantly greater in test (with MG) teeth than control (no MG) teeth. Mean forces to produce injury in test teeth decreased with resorbing root lengths. Deciduous incisors differed in injury type: subluxations and horizontal root fractures predominated in test teeth; lateral luxations and horizontal root fractures in control teeth. Predominant injuries in test and control permanent incisors were enamel infractions and subluxations. The magnitude of lateral luxation measurements of individual teeth was reduced significantly by mouthguard protection in both deciduous and permanent dentitions. The mouthguard tended to increase the mobility of the teeth it encompassed and, in some instances, promoted dentoalveolar injury of adjacent teeth. PMID- 9206376 TI - Cutting efficiency of K-files manufactured with different metallic alloys. AB - The aim of this study was to compare, in vitro, the machining efficiency of different triangular cross-section K-files made of nickel titanium (Nitiflex, Naviflex), titanium (Microtitane), and stainless steel (Flexofile, Flex-R). Ten instruments of each K-file from size 25 to 40 were tested. The cutting efficiency was assessed in a linear motion using an indentation caliper to measure the depth of grooves. The load applied (in grams) was equal to the ISO file size. Each file was allowed to do 100 back-and-forward movements. Files made of stainless steel were the most effective, in particular Flexofile. There were statistically significant differences between Flexofile and Flex-R in all sizes. In the group of nickel titanium instruments, Nitiflex was significantly more efficient than Naviflex in all sizes. The machining ability of titanium files was higher than that of Naviflex but lower than that of Nitiflex and stainless steel files. PMID- 9206377 TI - Bacterial penetration of the root canal of intact incisor teeth after a simulated traumatic injury. AB - One of the aims in treating traumatised teeth is to maintain the vitality of the pulp or allow conditions favourable for pulp revascularisation. However, infection of the pulp and root canal system may prevent this. A number of pathways have been proposed that allow bacteria to invade the root canal system, however most of these pathways cannot account for pulp infection in teeth that did not sustain injury to the periodontal attachment. Enamel/dentine cracks have been proposed as a portal for bacterial invasion of seemingly intact teeth and the aim of this study was to determine if bacteria could invade the root canal system after a simulated traumatic episode. Twenty intact and sound upper central incisors were chosen and prepared. One tooth was selected as a sterility control and the external crown surface of the remaining 19 teeth was subjected to infection with Streptococcus gordonii in a bacterial microleakage model. Over 7 days samples of growth media from the root canal system were taken and tested for bacteria. Sixteen of the teeth did not demonstrate bacterial invasion over the time frame. These teeth were then prepared for testing in a pendulum impact device and were subjected to a blow which did not fracture the crowns or dislodge the tooth from its simulated alveolus. The teeth were then prepared and tested in the bacterial microleakage model. After impact seven of the teeth demonstrated bacterial invasion of the root canal system (P = 0.002). These teeth were then reprepared for testing in the bacterial microleakage model. The crowns of five teeth, selected at random, were coated with two layers of light cured unfilled resin, the remaining two were used as positive controls. All the teeth coated with resin did not demonstrate bacterial invasion (P = 0.00), while the positive controls demonstrated invasion. The results suggested that enamel/dentine infractions were pathways for bacterial invasion of the root canal system of traumatised teeth. The application of unfilled resin to the anatomical crown prevented infection. PMID- 9206378 TI - Traumatic injuries to anterior teeth in Italian schoolchildren: prevalence and risk factors. AB - In the present study, the prevalence of traumatic injuries to anterior teeth in 824, 6- to 11-year-old, schoolchildren from Rome (Italy) and the relationship between injuries and predisposing factors were evaluated. Prevalence value of the study-population was 20.26%. This value is higher than those reported in surveys performed in Italian emergency dental services, but it is similar to those of retrospective studies from other countries. The highest prevalence was found among 9-year-old boys (33.69%); the M/F ratio was 1.64. The percentage of injuries with unknown cause (21.46%) was higher than that of other studies. This may be because most of injuries were slight (64.39% of injuries were enamel fractures) and it is likely that children and their parents were not worried about them, when they happened, so that they did not seek urgent dental care-this helps to explain the prevalence values of this and other retrospective studies, which are higher than those from emergency services-and, when interviewed, they did not remember the circumstances of the traumatic event. Using the Mantel Haenszel's Odds Ratios stratified for age and sex, injuries were related to individual predisposing factors (overjet larger than 3 mm: OR = 2.57, p = 0.0001, short upper lip: OR = 2.23, p = 0.0001 and upper medial incisor protrusion: OR = 3.95, p = n.s.), but not to children's trauma predisposing behaviour (OR = 0.92, p = n.s.). Serious injuries, however, happened to children without predisposing factors and were caused by strong impacts, suggesting that individual risk factors may not affect these type of injuries. PMID- 9206379 TI - Mental nerve paresthesia associated with a non-vital tooth. AB - Apical periodontitis is a common development associated with teeth with necrotic pulp. Although rare, some cases may present further complications, such as neuropathies in areas adjacent to the affected tooth. A case is described in which mental nerve paresthesia was associated with a non-vital mandibular premolar. Endodontic therapy resolved the paresthesia completely without further clinical complications. PMID- 9206380 TI - Accidental swallowing of endodontic instruments. AB - Swallowing or aspiration of a foreign body is a complication that may arise from any procedure in the oral cavity performed without the use of a rubber dam. Two cases of swallowed endodontic instruments and possible complications are presented. Emphasis is given in the discussion on the early diagnosis and treatment and primarily on prevention with the use of rubber dam, an absolute essential during endodontic treatment. PMID- 9206381 TI - Diffusion of resin monomers through human carious dentin in vitro. AB - The diffusion of 2-hydroxyethylmethacrylate (HEMA) and triethylene glycol dimethacrylate (TEGDMA) from light cured bonding resin-composite resin restorations through human carious dentin was investigated. Extracted human molar teeth with different degrees of caries were obtained from consenting donors. Teeth were classified into three groups according to caries severity (mild, moderate and severe) using subjective criteria. The outer carious lesions were then removed guided by a proprietary caries detector dye. Teeth with exposure of the pulp space after caries removal were excluded from the study. A polypropylene chamber was attached to the cemento-enamel junction of each tooth to contain 1 ml distilled water. Each cavity was restored with a HEMA containing bonding resin then a TEGDMA-containing resin composite. Water samples were retrieved over a time course and analyzed by high performance liquid chromatography. There was great variation between teeth in HEMA and TEGDMA permeability. The cumulative amounts released were of similar magnitude to those observed in non-carious teeth for the mild and moderately-severe groups. However, the cumulative amounts released were markedly greater in severely carious teeth than in those with moderate or mild caries. PMID- 9206382 TI - Immunohistochemical examination of the distribution of macrophages and CGRP immunoreactive nerve fibers in induced rat periapical lesions. AB - To clarify the role of calcitonin gene-related peptide (CGRP) in the development of periapical lesions, we examined the distribution of CGRP-immunoreactive (IR) nerve fibers and macrophages, and the behavior of bone tissues in experimentally induced rat periapical lesions by immunohistochemical and quantitative methods. Although no extensive changes were observed at 7 days after pulp exposure, CGRP IR nerve fibers increased in number until 28 days with a decrease thereafter. These neural changes were closely correlated with the alteration in number of macrophages except on day 7 when macrophages were significantly increased in number as compared with control rats. Tissue repair began to take place and a decrease in number of osteoclasts was observed when the density of CGRP-IR nerve fibers reached a peak. These results suggested that there might be a close relationship between macrophages and CGRP-IR nerve fibers and that CGRP-IR nerve fibers might participate in tissue repair in experimentally induced rat periapical lesions. PMID- 9206384 TI - Investigation of lay knowledge of the management of avulsed permanent incisors. AB - The prognosis of replanted avulsed permanent incisors depends largely on prompt and appropriate emergency management. The aim of this study was to investigate lay knowledge and attitudes in this respect. Postal questionnaires were sent to all physical education teachers, school nurses and secretaries, attendants in swimming baths and leisure centres and to 220 parents of teenage children in a defined area of North West England. The overall questionnaire response rate was 86.9%. Knowledge of methods of dealing with this problem was generally inadequate in both parents and the other groups. Although 53.6% of respondents claimed to have received first aid training only 3.1% could remember dental injuries being included. There was evidence that dental health education in this field can be effective, since the highest mean knowledge score was found in the 11.5% of respondents who recalled receiving advice from sources such as posters, magazines and newspapers. More than 80% of the respondents stated that they would not want to replant an avulsed incisor themselves, the main reason being lack of knowledge and training. It is suggested that there is a need for potentially effective dental health education in relation to this problem. PMID- 9206383 TI - Comparison of lipopolysaccharides from Bacteroides, Porphyromonas, Prevotella, Campylobacter and Wolinella spp. by tricine-SDS-PAGE. AB - Lipopolysaccharides (LPSs) of 11 bacterial strains from the type species of the genera Bacteroides (B. fragilis), Prevotella (Pr. melaninogenica), Porphyromonas (Po. gingivalis), Campylobacter (C. fetus subsp. fetus), and Wolinella (W. succinogenes), and from the type strains of B. distasonis, B. forsythus, B. ureolyticus, Po. levii, Po. macacae, and C. gracilis, were extracted with hot water-phenol (Westphal method). S-form LPSs, obtained from all organisms, were well resolved with tricine-sodium-dodecyl-sulphate polyacrylamide gel electrophoresis and visualized by silver staining. Lipid A was not stained. Also profiles from LPS of Escherichia coli, serotypes 0111:B4 and 055:B5, could be distinguished. While W. succinogenes showed a relatively short S-form LPS on electrophoregrams, the other bacteria, including B. fragilis, exhibited long ladder LPSs. Po. gingivalis displayed the largest number of bands and the longest O-chain. The long O-chain of this bacterium may be important for its virulence. PMID- 9206385 TI - Revascularization of traumatized teeth assessed by laser Doppler flowmetry: case report. AB - Infection of the pulp space in addition to the attachment damage of a traumatic injury to a tooth, results in serious complications and often tooth loss. Therefore, the prevention or treatment of root canal infection is a major consideration in these cases. In immature teeth, revascularization of a necrotic pulp is possible and highly desirable. Unfortunately, current sensitivity tests are poor indicators of revascularization, with the result that many pulps are removed unnecessarily. Laser Doppler flowmetry is an objective test of the presence of moving red blood cells within a tissue, which has been reported to be effective in the detection of tooth pulp vitality as well. A case is presented where an eight year old child severely luxated both maxillary central incisors. While only one of the incisors was weakly responsive to CO2 ice at 76 days after replantation, the laser Doppler flowmeter indicated that revascularization was occurring in both teeth at a much earlier time. Because of the laser Doppler readings, endodontic treatment was not initiated and the teeth developed normally. PMID- 9206386 TI - Experimental apexigenesis in baboons. AB - Apexification with calcium hydroxide is a routine procedure. However, some clinical reports suggest that root completion can occur by controlling the infection without use of a catalyst. The present study investigated the use of tetracycline treatment (in root canals) on root growth in immature teeth, rendered non-vital experimentally. Incisors in 3 young baboons were exposed and canals were left open. After 2 months all canals were cleaned and treated with either tetracycline or formocresol. Some canals in each group were filed. Animals were sacrificed after 6 months. Bacterial evaluations were done before placing medications, one week later and six months after that. The number of bacteria were reduced in all treatment groups. Root growth almost near completion was observed in more teeth treated with tetracycline than in the formocresol group. PMID- 9206387 TI - Comparison of two desensitizing agents for the treatment of cervical dentine sensitivity. AB - Numerous desensitizing agents have been utilized in an effort to alleviate the discomfort associated with cervical dentine sensitivity (CDS). Recently several new tubule-occluding and sealant systems have been marketed for treatment. The aim of this study was to compare two desensitizing agents (ALL-BOND 2 and Butler Protect) in a 3-month clinical study. Ten subjects (6F; 4M mean age 45.1 years (SD 8.81) who had provided voluntary written informed consent participated in a single-blind 3-month clinical study. Subjects were evaluated for tactile (Yeaple probe) and air sensitivity (dental air syringe) together with subjective perception of pain (VAS scores) at 0.5 min, 1, 2 and 3 months. There was an overall trend in reduction of CDS over the study period in all group with no significant differences detected between groups. The results suggested that while subjects reported overall reductions in sensitivity levels, this may not necessarily be substantiated when assessed objectively. Furthermore, there appeared to be a strong placebo effect in this study. PMID- 9206388 TI - Mouth guard for athletes during orthodontic treatment. AB - Mouth guards have been proven to greatly reduce the number and severity of traumatic dental injuries to participants in contact sports. However, there are some difficult problems in making mouth guards for athletes with orthodontic appliances on their teeth. We developed a method of making mouth guards by using a coating material for such athletes to prevent dental and oral sports injuries and to protect the orthodontic appliances. This mouth guard fits well and does not damage the appliances. PMID- 9206389 TI - Endodontic management of a rare combination (intrusion and avulsion) of dental trauma. AB - Combined trauma involving intrusive luxation of one tooth and avulsion of another is rare. A case is presented involving the endodontic management of two traumatised maxillary central incisors, one of which was intrusively luxated and the other avulsed. Spontaneous re-eruption of the intruded tooth occurred, thereby avoiding the need to further traumatise the periodontal ligament with either orthodontic or surgical repositioning, and allowing endodontic therapy to be carried out uneventfully. Endodontic therapy of the avulsed tooth was completed and its prognosis is considered good. PMID- 9206390 TI - Three independent canals in the mesial root of a mandibular first molar. AB - Endodontic therapy was performed in a mandibular first molar with three canals in the mesial root. The mesiobuccal and the mesiolingual canals were found in their normal locations. The third was located in the middle of the distance between the other two. Radiographically, it ended in its own distinct foramen. Many reports deal with three orifices in the mesial root, but very few describe three independent canals, indicating a rare anatomical configuration. To locate the third possible intermediate canal it was suggested to reduce the mesial dentinal wall of the pulp chamber after instrumenting the main two canals. PMID- 9206391 TI - Evaluation of three slip fit hexagonal implants. AB - Three external hexagonal implant systems were evaluated for machining tolerances and component fit. Mean implant hexagonal flat to flat size dimension varied from 2.685 to 2.700 mm, and a close correlation among corresponding analog hexagonal flat to flat dimensions for each system was noted. Rotational misfit for implant abutment matings varied from 1.6 to 5.3 degrees. Torsional strength for four different matings ranged from 122.7 to 175.8 N-cm. The data presented suggest a trend by some implant manufacturers toward improved tolerances, accuracy, fidelity, and consistency of implant components. PMID- 9206393 TI - The locking taper attachment for implant abutments: use and reliability. AB - The locking taper abutment attachment for implants is a one and one-half-degree tapered post that relies on friction rather than a screw for retention of the abutment to the implant. A retrospective analysis of 1,757 consecutively loaded implants was made to determine the reliability of the locking taper abutment. Five hundred and four implants restored over a 7-year period from one practice and a total of 1,253 implants restored over 4 years from 2 practices were reviewed. Nine abutment posts fractured for a failure rate of 0.05 percent. Thirty one (1.7 percent) of the abutments loosened but were reinserted without further problems. The technique for use of the locking taper implant abutment attachment is described. PMID- 9206392 TI - Bone augmentation and biopsy: clinical report. AB - A mixture of demineralized freeze-dried cortical bone and beta-tricalcium phosphate covered by titanium-reinforced expanded polytetrafluoroethylene membranes was used for bone augmentation in an accident case in which a young female patient suffered anterior maxillary damage. Hydroxyapatite-coated implants were placed in the newly formed bone-like material before prosthodontic reconstruction. A biopsy suggests the bone-like material was essentially normal lamellar and woven bone. PMID- 9206394 TI - In vitro mineralization and implant calcium phosphate-hydroxyapatite crystallinity. AB - Biological dissolution of implant calcium phosphate coatings release local concentrations of divalent ions, which may influence mineralization. The objective of this study was to determine the effects of calcium phosphate release from coated commercially pure titanium discs using a bone-like cell culture bioassay. Sandblasted discs were prepared with or without hydroxyapatite crystallinities (50, 75, and 90 percent). Samples of each coating were randomly assigned and either preincubated for 24 hours with media or not before the addition of cells (2200/ mm2). Cultures were grown for 72 hours in culture medium containing 0.5 microCi/mL45 Ca. After rinsing, the remaining calcium phosphate surface was dissolved and counted. Three independent trials were performed. Results indicated proliferation was not altered as a function of crystallinity (P > 0.05) among any of the groups. However, a significant (P < 0.01) inverse relationship was found for biologically mediated mineralization as a function of calcium phosphate crystallinity. Low crystalline surfaces (nominally 50 percent) had the highest level of mineralization, with 75 percent crystalline surfaces being intermediate and 90 percent crystalline samples having the lowest amount of relative mineral formation. Mineralization only occurred on sandblasted commercially pure titanium upon supplementation of the growth medium with an organophosphate (beta-glycerophosphate), although this was less than on culture plastic. The results suggest calcium phosphate dissolution, as a function of implant coating crystallinity, can alter biological mineralization and may be one means in which enhanced mineral formation occurs around calcium phosphate-coated dental implants. PMID- 9206395 TI - Evaluation of the surface integrity of hydroxyapatite-coated threaded dental implants after insertion. AB - The surface characteristics and changes of hydroxyapatite-coated threaded dental implants after insertion into bone (veal calf ribs) with similar cortical and medullary characteristics to the human maxilla and posterior mandible (type III or IV bone) were studied. Hydroxyapatite-coated threaded implants from six vendors were coded A, B, C, D, E, and F. Four implants from each vendor were subjected to conventional placement following the manufacturer's instructions, placement without tapping of the osteotomy site, or evaluated as controls. The implants were recovered atraumatically and examined by scanning electron microscopy at x 35 and x 100 magnification. Photomicrographs were examined and graded by two independent examiners. Statistically significant differences were found using Kruskal-Wallis ANOVA and Wilcoxon Signed Ranks tests in surface integrity between conventional and nontapped treatment for groups C and D; between conventional placement and controls for groups A, B, and F; between nontapped placement and controls for groups A, B, C, D, and E; and among the manufacturers with respect to conventional placement for groups A, B, and E as compared with the other groups (P < 0.05). These findings suggest that surface changes of hydroxyapatite-coated threaded implants may occur during placement, particularly in undersized and untapped osteotomy sites. The changes could result in differences in integration and performance of some implant systems. PMID- 9206397 TI - New Directions. Calcitek 1996 International Symposium. San Diego, California, August 1-3, 1996. Abstracts. PMID- 9206396 TI - Atypical incisive nerve: clinical report. AB - A 68-year-old Caucasian man complained of discomfort from two implants placed in the anterior mandibular area. Examination of cross-sectional computerized tomography images revealed the passage of a distinct structure, surrounded by cortical bone, which extended from the right mental foramen anteriorly and passed through the lower part of the right and left implants. The structure, which was identified as the incisive nerve, also emerged from the left mental foramen and advanced toward the midline. This nerve plexus is usually delicate and not detectable by conventional radiography or computerized tomography. The possibility of nerve variations in the anterior region of the mandible must be considered to prevent possible trauma to the adjacent tissues when placing implants. PMID- 9206398 TI - Twenty-five years--where courage has led the ICOI. PMID- 9206399 TI - Repair of a transmandibular implant: clinical report. AB - The use of many currently available implant systems may be limited in patients with severe mandibular atrophy because of the lack of available bone in the symphyseal region. The transmandibular implant is a rigid box-frame structure that can distribute masticatory forces along a severely atrophied mandible with as little as 4 mm of vertical bone with an implant-borne prosthesis. Although the success rate of the transmandibular implant is favorable as compared with other systems, complications can occur after placement, including infection, partial extrusion, and fracture of a post. A case report of fractures of two posts at the necks and the bar approximately 18 months after placement is presented. PMID- 9206400 TI - The abutment seating jig: a prosthodontic implant adjunct. AB - Rapid, accurate seating of screw-retained implant abutment heads, where timing is controlled by internal or external hex designs, can be readily accomplished with individual, custom-cast abutment head location devices. The devices are especially useful when the abutment head-implant body complex is to be permanently cemented. The use and design of abutment seating jigs for single tooth implants and completely implant or implant and natural tooth-supported prostheses are described. PMID- 9206401 TI - Surface roughness of titanium on bone morphogenetic protein-2 treated osteoblast cells in vitro. AB - Surface topography plays a critical role in the interaction of dental implants with adjacent tissues. No statistical differences in oxide composition and surface contamination were observed between 600 grit and polished titanium surfaces. The expression of osteoblast phenotype was enhanced when osteoblast progenitor cells (2T9) were stimulated with bone morphogenetic protein-2 on polished and 600 grit titanium surfaces. Bone morphogenetic protein-2 stimulated phenotypic expression on 600 grit titanium surfaces was marked by prolonged alkaline phosphatase specific activity and more rapid osteocalcin production as compared with the polished titanium surfaces. Because the surface area of the 600 grit titanium surface was shown to be 8 percent greater than that of the polished titanium surface, it is possible that increased surface area played a role in the enhanced expression of the osteoblast phenotype. PMID- 9206402 TI - Comparison of screw retention of nine abutment systems: a pilot study. AB - Fracturing or loosening of abutment and retaining screws is a frequent occurrence. This study was designed to evaluate the retaining ability of nine abutment systems. Abutment and retaining screws were secured to the appropriate implant using a digital torque gauge, with the abutment screws placed at 30 N-cm and the retaining screws at 10 N-cm. Removal torque was recorded three times for each sample 10 minutes after initial placement, 20 minutes after retightening, and 24 hours after retightening. There was no statistical difference in removal torque relative to time or repeated tightening. Of the nine abutment systems, three were statistically less retentive as compared with the most retentive system. The retaining screws were less retentive as compared with the abutment screws. For maximum retention the elimination of retaining screws from the restorative design and using 30 N-cm of torque for the abutment screws is suggested. PMID- 9206403 TI - Implant pathology associated with loss of periapical seal of adjacent tooth: clinical report. AB - A mandibular cuspid adjacent to two implants placed in the incisor region redeveloped periapical pathosis as a result of the inadvertent removal of the gutta-percha seal during post preparation. The root end inflammatory process communicated with the surface of an adjacent implant, resulting in endodontic implant pathosis and subsequent removal of the implant. The osteotomy site healed uneventfully and almost complete osseous repair was observed after five months. PMID- 9206404 TI - Implants Advance into the 21st Century. The International Congress of Oral Implantologists World Congress XVI in Association with the Sociedad Espanola de Implantes. Madrid, Spain, October 18-20, 1996. Abstracts. PMID- 9206405 TI - Implant Prosthodontics: Practical Techniques for the Implant Team. The American Academy of Implant Prosthodontics 15th Annual International Symposium in Association with Medical University of South Carolina College of Dental Medicine. Atlanta, Georgia, November 8-10, 1996. Abstracts. PMID- 9206406 TI - Penetration of smeared or nonsmeared dentine by Streptococcus gordonii. AB - The purpose of this study was to investigate the penetration of smeared and nonsmeared dentine by Streptococcus gordonii. Prepared human roots, grouped as either nonsmeared or smeared, were immersed in a suspension of S. gordonii cells for 3 weeks. The roots were then prepared for scanning electron microscopy and histological analysis. Dentine discs prepared from coronal dentine were grouped similarly. Using a fluid filtration apparatus, the hydraulic conductance (Lp) of each disc was determined before and after incubation with bacterial suspension. Scanning electron microscopy evaluation of the roots following infection with bacteria showed no change in the smear layer (P < 0.0001). Histological sections revealed that bacterial penetration of all the nonsmeared samples had occurred, while nine out of 10 smeared samples showed no bacterial penetration (P < 0.0001). The Lp of the nonsmeared discs was significantly reduced by 42% (P < 0.0001) after bacterial penetration. However, the smeared samples revealed a 1% reduction in Lp which was not significant (P > 0.05). The results suggest that dentinal smear layers are an effective barrier to dentinal tubule invasion by S. gordonii. PMID- 9206407 TI - Permeability, morphologic and temperature changes of canal dentine walls induced by Nd: YAG, CO2 and argon lasers. AB - The permeability, temperature and morphologic changes of the wall of the root canal induced by Nd:YAG, CO2 and argon lasers were studied. The changes were evaluated according to the presence or absence of a smear layer. Root canals of 140 human single-rooted teeth were enlarged using a step-back technique. Permeability was evaluated by the extent of methylene blue dye penetration into the tubules. Temperature changes were measured using a thermovision system, and morphological changes were evaluated by scanning electron microscopy. Laser energy was delivered into the canal by means of a flexible optical fibre or metal tip. There were statistically significant differences in permeability between lased groups with and without a smear layer in the cervical third of the root canal following lasing. In the middle third of the root canal, all three laser types induced permeability increases in groups with a smear layer. In the apical third, statistically significantly decreases in permeability were observed among CO2 laser and Nd:YAG compared with control group (P < 0.01). Rises in temperature ranged from a minimum of +10.1 degrees C (CO2 laser) to a maximum of +54.8 degrees C (argon laser). All three laser devices appeared capable of producing a glazed-like surface and craters. PMID- 9206408 TI - The effect of tubular penetration of root canal sealers on dye microleakage. AB - An in vitro study of a possible correlation between penetration of dentinal tubules by four root canal sealers and microleakage of external fluids into the canal was done using a dye leakage method and scanning electron microscopy. The root canals of 45 teeth were instrumented and the smear layer removed prior to obturation of root canals with gutta-percha and one of four sealers: Diaket, Endomethasone, CRCS or Ketac-Endo. The extent of leakage was scored after immersion in India ink for 72 h. The same specimens were also used for scanning electron microscopic evaluation. There was a statistical difference in leakage patterns between the groups (P < 0.05). Diaket had lower microleakage scores than the other sealers (P < 0.05). When the scores for penetration of sealers into the tubules were analysed, Ketac-Endo demonstrated the least penetration (P < 0.01). There appeared to be a converse relation between tubular penetration and dye leakage, but the correlation was not statistically significant (P > 0.05). PMID- 9206409 TI - The use of a modelling technique to investigate the root canal morphology of mandibular incisors. AB - The prime objective of this study was to develop and test a method of modelling the dental root canal system using a vinyl resin, type UCAR-VYHD. It also aimed to verify if the resin models allow a three-dimensional evaluation of those features of the pulpal morphology of human permanent mandibular incisor teeth, which might be associated with periodontal disease. One hundred and eleven freshly extracted teeth were studied. Pre-extraction clinical records and periapical radiographs were used to deduce the presence or absence of periodontal disease. Each extracted tooth was radiographed in two perpendicular planes before being opened by conventional orthograde endodontic technique, and photographed in a scanning electron microscope. The patent pulp space was then obturated with the vinyl resin. Scanning electron photomicrographs of the resin replicas and of the teeth were compared with the earlier radiographic images to correlate the morphological findings. The numbers of root canals and apical foramina were recorded. This study showed that in mandibular incisors with periodontal disease there was a relatively low incidence of lateral canals (7.2%). The modelling technique was found to be reliable, reproducible, and provided sufficient detail to allow three-dimensional analysis of root canal morphology. PMID- 9206410 TI - Plasma catecholamine and haemodynamic responses to surgical endodontic anaesthetic protocols. AB - The effects of varying clinically relevant patterns of anaesthetic vasoconstrictor combinations used for periradicular surgery on plasma concentrations of catecholamines and haemodynamic responses was studied in the canine model. Five mongrel dogs were anaesthetized with sodium pentobarbitol. A femoral cannula was inserted to measure central blood pressure and an ECG was used to monitor heart rate and any associated arrhythmias. Femoral venous blood samples were drawn before initial injection and at 3 and 10 min after injections. Plasma catecholamine concentrations were determined using high pressure liquid chromatography (HPLC). Injection protocols used three time periods, 30, 60 and 90 s, with solutions containing 1:100,000 and 1:50,000 adrenaline. No significant changes in heart rates or presence of arrhythmias were noted over the experimental protocol. Catecholamine levels in pico moles mL-1 were within the normal range at the 3-min sample level. At the 10-min sample time there was a more erratic range of concentrations, with most samples within the normal range. This may have been due to endogenous release of catecholamines in specific animals. The data identified trends in both the haemodynamic parameters and plasma catecholamine levels that can legitimately support the careful use of higher levels of a vasoconstrictor when patient profiles and surgical needs dictate. PMID- 9206411 TI - Root-end cavity preparation using the MicroMega Sonic Retro-prep Tip. SEM analysis. AB - The objective of this laboratory study was to compare root-end cavities prepared with sonic Retro-prep tips in a MM1500 Sonic Air handpiece with those created by burs in a conventional handpiece. A total of 80 single-rooted extracted human teeth with mature apices and straight canals were included in the study. Four groups of 20 extracted teeth were prepared as follows: I, a 3-4 mm root-end resection perpendicular to the long axis of the root, with a size 40 sonic Retro prep tip creating an apical cavity 3 mm into root canal system; II, a 45 degrees bevel of the root-face removing a 3-4 mm root segment and root-end preparation as per group I; III, root-end resection as per group I, with an apical cavity prepared using a size 010 inverted cone bur 3 mm down the long axis of the root; IV, resection as per group II, followed by an apical cavity preparation with a size 010 inverted cone bur 3 mm into the root canal system. The apical root portion and root-end cavities were replicated and prepared for SEM analysis at x 20 and x 80 magnification. The degree of chipping associated with the margin of the root-end cavities, as evaluated with a standard grading system, and the incidence of root-face cracks were noted. Marginal chipping of root-end cavities prepared using sonic instrumentation was significantly worse than that produced by burs (P < 0.001). Perpendicular root-end resections showed significantly better scores than bevelled root-end resections (P < 0.005). The incidence of root-face cracking was low with no significant difference between the experimental groups. PMID- 9206412 TI - Cutting efficiency of Hedstrom, S and U files made of various alloys in filing motion. AB - The cutting efficiency for sizes 25 and 35 stainless steel Hedstrom S and U files from 10 manufacturers, and titanium-alloy Hedstrom S and U files from five manufacturers, tested for linear (filing) motion under standardized conditions were determined. Special plastic samples having well-defined abrasive properties were used as the substrate and constant pressure was applied until the instruments were blunt. The depth of the groove achieved by filing was used to measure cutting efficiency. For both sizes there were significant differences in the cutting efficiency of files made by the various manufacturers (P < 0.001). Hedstrom files made of stainless steel, made by VDW, gave the best cutting efficiency for sizes 25 and 35. Overall, under the conditions of this study. Hedstrom files had better cutting efficiency than S or U files. Likewise, stainless steel files provided better cutting efficiency than instruments made of titanium alloys. PMID- 9206414 TI - Comparison of three instrumentation techniques in the preparation of simulated curved root canals. AB - Forty-five acrylic blocks with simulated curved canals were divided into three groups of 15 blocks each. In group 1 the canals were instrumented with ultrasonically energized K-files (UEF) using a piezoelectric ultrasonic device. In group 2 the canals were prepared with ProFile 0.04 Taper Series 29 Rotary Instruments (PRI) in conjunction with a low-speed high torque handpiece. In group 3 (control) the canals were hand instrumented (HI) with conventional K-type files using a standard push-pull circumferential technique. The efficiency of these techniques for preparing the simulated canals were compared by measuring the amount of transportation of the prepared canals at different levels from the working length using a double exposure photographic technique. A statistical analysis was used to indicate any significant difference among groups. The results showed that the use of PRI provided well-centred and more tapered preparations. Conversely, the use of UEF and HI resulted in frequent alteration of the original curvature, showing transportation at different levels from the working length. PMID- 9206413 TI - Intra-canal cutting ability of MM1500 files. AB - The aims of this study were to develop a model system capable of monitoring lateral forces during root canal preparation and to measure the cutting ability of files activated by the MM1500 sonic handpiece. Forces were monitored by a calibrated model system which utilized a combination of spring steel beams fitted with strain gauges, these were interfaced through two strain gauge amplifiers to an x-y recorder. Single rooted canine teeth (n = 36; 32 experimental, four control) were mounted in a two-part acrylic mould (which was an integral part of the model system) prior to sectioning horizontally 11 mm from the tooth apex. A 2(4) full factorial experiment with two replications was performed. Four variables were selected for evaluation, load (50 and 100 g), power (air inlet ring half or fully open), file type (Heliosonic or Shaper) and stroke rate (1 or 2 cycles per second). A new file (size 25) was used for 1 min in each canal with water irrigation. The control group was not instrumented. The cross-sectional root canal area was measured before and after instrumentation using image analysis and increase in area was used as an indication of cutting ability. The results showed that the increase in load, power and the Shaper file all produced a significant increase in cutting ability (ANOVA, P < 0.001). However, stroke rate was not found to have a significant effect (P > 0.05). None of the interactions between the variables were significant and there was no significant difference in the control group (P > 0.05). In conclusion, this work has developed a model system to monitor lateral forces and has shown that instrument design and operator usage affect dentine removal from a root canal wall. PMID- 9206415 TI - Susceptibility of Actinomyces israelii to antibiotics, sodium hypochlorite and calcium hydroxide. AB - Actinomyces israelii has been repeatedly implicated as a cause of failure of endodontic therapy. This study investigated the antimicrobial effect of antibiotics as well as intracanal medicaments, sodium hypochlorite solution and calcium hydroxide, on this important pathogen. Growth of A. israelii was inhibited by low concentrations of antibiotics, yet high concentrations were not bactericidal for A. israelii over 1 week. When A. israelii was exposed for 2-6 weeks at concentrations equivalent to clinical serum levels, the antibiotics were lethal. The results reveal a species-specific antibiotic tolerance for A. israelii. Both sodium hypochlorite solution and calcium hydroxide were found to be highly effective in killing A. israelii. PMID- 9206416 TI - The sealing ability of Thermafil obturators assessed by four different microleakage techniques. AB - An in vitro investigation was performed to assess the extent of apical dye leakage in relation to root fillings made by Thermafil obturators and the lateral condensation of gutta-percha in extracted human teeth under conditions of passive dye penetration, centrifugation, a vacuum technique and an increased air pressure technique. One hundred and twenty-eight extracted teeth were selected and prepared. The specimens were allocated into eight closely matched experimental groups. Four groups were obturated with Thermafil obturators and four with the lateral condensation technique. Microleakage was assessed after exposing one group of specimens from each of the obturation techniques to each of the four dye penetration systems using India ink as the leakage detector. The teeth were demineralized and cleared prior to examination and the maximum dye penetration for each specimen was recorded. The statistical analysis on transformed data revealed no significant differences between the four microleakage techniques and no differences between the two obturation techniques. PMID- 9206417 TI - Effect of various irrigant and autoclaving regimes on the fracture resistance of rubber dam clamps. AB - Rubber dam clamps are known to break during clinical use in endodontics. This in vitro study examined some of the variables which may contribute to the fracture. Stainless steel rubber dam clamps were subjected to various cleaning and autoclaving regimes and exposure to various solutions of sodium hypochlorite (NaOCl). Each clamp was examined after four cycles of cleaning and exposure to NaOCl. During environmental exposure to NaOCl, the clamp was stressed over a perspex rod to simulate placement onto the crown of a tooth. Clamps were examined after each test cycle visually and microscopically, or immediately after breakage. Results suggested that the fractures were because of a stress corrosion cracking phenomenon. There was evidence of intergranular and transgranular cracking of the metal. Corrosion spots were seen on the surface of the clamps and fracture occurred mainly through these spots. A number of recommendations to reduce breakage of clamps have been suggested. PMID- 9206418 TI - Haemostasis in periradicular surgery. AB - The successful performance of endodontic surgical procedures is predicated on many factors. However, the ability of achieve sustained tissue haemostasis in the surgical site is crucial to the performance of these procedures. This achievement improves vision in the surgical site, minimizes surgical time, enhances the surgical procedures (root-end resection, preparation and filling), and reduces surgical blood loss, postsurgical haemorrhage and postsurgical swelling. A multitude of materials have used in dentistry and medicine to achieve both generalized and localized haemostasis, many without full assessment of their biological implications. The purpose of this paper is to provide a thorough and critical review of these materials from the perspective of surgical endodontics, highlighting their development, application and potential role in achieving proper haemostasis. PMID- 9206419 TI - Time-course and risk analyses of the development and healing of chronic apical periodontitis in man. AB - Roots with and without preoperative chronic apical periodontitis were root canal treated and followed clinically and radiographically yearly for up to 4 years. Of 732 roots treated, 599 (82%) were available for evaluation at one or several recalls. Chronic apical periodontitis (CAP) was recorded with the periapical index scoring system. CAP developed in 29 of 473 (6%) of teeth without preoperative signs of disease, whereas 111 of 126 (88%) initially diseased roots showed signs of healing. The rate of healing CAP and the rate of emerging CAP were calculated, and analyses of event occurrence each year of observation were performed. Peak incidence of healing or emerging CAP was at 1 year in both instances. Risk assessments at 2, 3, and 4 years did not indicate an added risk of filled roots developing CAP during this period. Complete healing of preoperative CAP in some instances required 4 years for completion, while signs of initiated, but incomplete, healing were visible in at least 89% of all healing roots after 1 year. Risk analyses may provide relevant information in addition to or in substitution for success/failure analyses. PMID- 9206421 TI - Microbiological root canal sampling: diffusion of a technology. AB - Diffusion is the process whereby a technology enters and becomes part of the health-care system. In the present study, diffusion of microbiological root canal sampling (MRS) among general dental practitioners within the city of Gothenburg was observed for 25 years, from the establishment of the Laboratory of Oral Microbiology in 1966. Laboratory records at 5-year intervals were analysed and adopters were categorized as 'occasional' (1-2 samples/year), 'selective' (3-10) or 'regular' (> 10) samples. The diagnostic accuracy was assessed by analysing the results of culturing 574 samples referred in 1986. The acceptance rate varied between 2.9 and 5.1% except in 1986 when 10.1% of the practitioners in the area referred root-canal samples. While a minority of the adopters used MRS as a standard procedure, the strategy appeared to be directed towards selection of special cases. The results of culturing revealed a predominance of facultatives. Although evidence of contamination was found, the practitioners frequently appeared to produce valid microbiological samples. PMID- 9206420 TI - Variation in the microleakage produced by four different techniques in root fillings in a simulated root canal model. AB - An in-vitro investigation was performed to assess the extent of apical microleakage in simulated canals in clear resin blocks. Four different methods of producing dye penetration were employed: passive dye penetration alone, centrifugation alone, vacuum plus passive dye penetration, and increased pressure plus passive dye penetration. Forty simulated root canals were prepared and obturated. The specimens were randomly allocated into four groups and each group was subjected to a different leakage technique using methylene blue dye as the detector. The maximum apical dye penetration observed through each of the four faces of the blocks was recorded. There were large variations in the leakage recorded in each individual specimen and between the specimens within each group. There were statistically significant differences between passive dye penetration and each of the other three techniques. There was no significant differences between the centrifugation, vacuum and pressure techniques. PMID- 9206422 TI - Reasons for dentists' acceptance or rejection of microbiological root canal sampling. AB - Microbiological root canal sampling (MRS) has been found to be used by only a few Swedish general dentists. The present study addresses the reasons for their acceptance or rejection of the technology. A questionnaire was mailed to 240 general dentists practising within the city of Goteborg. The questionnaire concerned certain practice characteristics and attitudes to MRS. The data showed that MRS is mainly performed by dentists working with adult patients in private practice. The technology is rarely used routinely, but is applied in selected cases. The main reason for non-adoption seems to be a perceived lack of relative advantage over conventional treatment strategies. Furthermore, opinions regarding the complexity and observability of the technology appear to influence acceptance significantly. PMID- 9206423 TI - Factors affecting the cutting ability of sonic files. AB - The aim of this study was to investigate the cutting ability of sonic files. A model system was developed and the following variables evaluated: file type. Heliosonic or Shaper; file length, 21 or 29 mm; power, air inlet ring opening of half or fully open; stroke length, 2 or 4 mm; stroke rate, one or two cycles per second; and load 50 or 100 g. A 2(6) full-factorial analysis with two replications into the effect of the above variables on the cutting ability of the MM1500 sonic instrument was performed. A new size 25 file was used for each cut, together with water irrigation, and the substrate used was 1-mm thick sections of bovine bone. The differences between the variables were significant (ANOVA, P < 0.001). However, examination of the F-values showed that the most significant variable to affect cutting was load, followed by power, file type, stroke length and stroke rate, with the least significant variable being file length. The most significant interaction was between rate and length of stroke. An increase in stroke rate from one to two cycles per second at a stroke length of 2 mm produced an increase in cutting for both the Heliosonic and Shaper files. However, at the longer stroke length of 4 mm, the same increase in rate resulted in a decrease in cutting for the Shaper files. Therefore, it is suggested that operators should press the file against the canal wall and move it slowly to maximise cutting. PMID- 9206424 TI - Detection of epidermal growth factor receptor in inflammatory periapical lesions. AB - Epithelial cell proliferation is often observed in periapical lesions of endodontic origin. The mechanisms which stimulate the epithelial cell rests of Malassez to proliferate are not understood fully. Fifteen inflammatory periapical lesions (10 granulomas and five cysts) obtained from periapical surgery and six additional periapical lesions (four granulomas and two cysts) collected from extracted teeth were examined using immunohistochemical staining and 125I-EGF (epidermal growth factor) binding assay to detect the presence of epidermal growth factor receptor. The results indicated that the periapical lesions without epithelial cell proliferation had a weak immunoperoxidase staining or low specific binding of 125I-EGF. In contrast, the periapical lesions with epithelial cell proliferation and cyst formation exhibited a strong immunoperoxidase staining in the epithelial cells or high specific binding of 125I-EGF. PMID- 9206425 TI - A comparison of bending and torsional properties of K-files manufactured with different metallic alloys. AB - The purpose of this study was to compare bending (bending moment) and resistance to fracture by twisting (torsional moment and angular deflection) of triangular cross-section K-files made of either nickel titanium (Nitiflex, Naviflex), titanium (Microtitane) or stainless steel (Flexofile, Flex-R). A total of 200 files were tested, 10 instruments for each type from size 25 to 40, according to ANSI/ADA specification no. 28 and ISO reference no. 3630. Files made of nickel titanium, especially Nitiflex, were the most flexible. Stainless steel instruments presented a higher bending moment than files made of nickel titanium and titanium, particularly Flex-R sizes 35 and 40. With regard to resistance to fracture, measured by angular deflection at the failure point, Flexofile followed by Flex-R were the most resistant to fracture and Nitiflex were the least resistant. Differences in angular deflection among file groups were greater than those for torsional moment. Thus, it seems that angular deflection is a more specific measurement for assessing resistance to fracture by twisting. PMID- 9206426 TI - Phantom tooth pain: a diagnosis of exclusion. AB - This case report addresses the difficult diagnosis of phantom tooth pain (PTP). This is a syndrome of persistent pain in the teeth and oral structures following pulp extirpation, extraction, or rarely, an inferior alveolar nerve block. The incidence of this often misdiagnosed condition is estimated to be 3% of the population undergoing pulp extirpation, and is similar to phantom limb pain. The diagnosis of PTP is discussed here as a diagnosis of exclusion, after numerous interventions. Various treatment modalities are outlined and the use of non traditional pharmacological approaches for pain reduction are discussed. PMID- 9206427 TI - Associations of endodontic symptoms and signs with particular combinations of specific bacteria. AB - Significant associations have been reported between (a) specific bacterial species isolated from root canals and (b) between individual bacterial species and endodontic symptoms and signs. The prime objective of this study was to determine whether particular combinations of specific bacteria are associated with individual endodontic symptoms and signs. Seventy root canals were investigated microbiologically taking care to maintain the viability of obligate anaerobes, which accounted for 64% of the total species isolated, including Peptostreptococcus micros, Prevotella melaninogenica, Prevotella oralis, Eubacterium aerofaciens, Eubacterium lentum, Fusobacterium nucleatum, Prevotella buccae and Prevotella intermedia. Significant associations were found between individual clinical features and the following pairs of species: (a) pain (37 cases) and Peptostreptococcus spp./Prevotella spp., Peptostreptococcus spp./Prevotella melaninogenica, Pstr. micros/Prev. melaninogenica (all P < 0.01); (b) swelling (23 cases) and Pstr. micros/Prevotella spp. (P < 0.01); (c) 'Wet' canal (57 cases) and Prevotella spp./Eubacterium spp. (P < 0.01), Peptostreptococcus spp./Eubacterium spp. (P < 0.05). Thus data from this investigation suggests that statistically significant associations exist between individual endodontic symptoms and signs and particular combinations of specific bacteria. PMID- 9206428 TI - Apical and coronal seal of roots obturated with a dentine bonding agent and resin. AB - The efficacy of sealing root canal systems with a dentine bonding agent and resin cement was compared with that of a glass ionomer sealer. Scanning electron microscopic examination of the bonding agent-dentine interface was also performed. The root canals of 50 single rooted teeth with mature apices were prepared chemomechanically and the smear layer removed with 17% REDTA and 5.25% NaOCl. Specimens were divided into two groups of 24 teeth each. One group had the dentine conditioned with a 10:3 citric acid-ferric chloride solution and obturated with the dentine bonding agent and resin, radiopaque C & B Metabond. The other group was obturated with the glass ionomer sealer, Ketac-Endo and a single cone of gutta-percha. After immersion in Indian ink for 90 h, the teeth were cleared and the quality of the apical and coronal seal was assessed using India ink dye penetration. There was a significantly better seal in both the apical and coronal directions when using the dentine bonding agent and resin obturation material. Scanning electron microscopic examination of the demineralized dentine and the C & B Metabond interface revealed the presence of the characteristic hybrid layer along with microtags of resin penetrating deep into the dentine tubules. PMID- 9206430 TI - Clinical uses of rectified turpentine oil. AB - Rectified turpentine oil can be used to soften or dissolve gutta-percha in the root canal space to facilitate endodontic retreatment or preparation of space for a post. For endodontic retreatment, the turpentine oil can be heated to 71 degrees C which significantly increases its ability to dissolve gutta-percha. For removing only a portion of the gutta-percha while leaving the rest intact, as for the preparation of space for a post, the turpentine oil is delivered to the canal at body temperature (37 degrees C). PMID- 9206429 TI - Periradicular healing in response to Diaket root-end filling material with and without tricalcium phosphate. AB - The healing of the periradicular tissues was evaluated when the polyvinyl resin Diaket with and without tricalcium phosphate was used as surgical root-end filling material. Non-surgical root canal treatment was performed on 56 mandibular premolar roots in mongrel dogs. Following root-end resection, root-end cavity preparations were filled with Diaket, the comparative material, or Diaket in combination with tricalcium phosphate, the experimental material. Postsurgically, healing of the tissues adjacent to the filling materials and in the surrounding surgical site were evaluated at 30 and 60 days. There was virtually no statistically significant difference between the experimental and comparative group at or within the 30- or 60-day period with regard to inflammation, connective tissue formation, root-end encapsulation, cementum formation, or bone apposition. Findings suggest that cementogenesis occurred over both materials. The overall healing of the periradicular tissues was favourable. PMID- 9206431 TI - The use of a radiopaque contrast medium in endodontic radiography. AB - A series of in vitro studies were carried out to investigate the use and application of a radiopaque contrast medium in conventional periapical dental radiography for the diagnosis and evaluation of root canal systems. The water soluble radiopaque contrast medium was introduced into the root canals of 30 first permanent maxillary and 30 first permanent mandibular molar teeth. The radiographic images of these teeth with and without radiopaque contrast medium in the root canal systems were compared and contrasted. Further comparisons were made with the same teeth rendered transparent. The results indicate that by standardizing the diagnostic criteria the inter-examiner reliability was in good agreement; it was independent of the radiographic technique used. The validity of the radiographs was enhanced by the use of the radiopaque contrast medium. The results confirm that, with the use of a radiopaque contrast medium, images of root canal systems are easier to read and interpret than plain radiographic images of root canal systems. The use of radiopaque contrast medium in endodontic radiography may be a valuable aid in the diagnosis and evaluation of root canal systems. This system would complement rather than replace plain radiography. PMID- 9206432 TI - An in-vitro study of smear layer removal and microbial leakage along root-canal fillings. AB - The objective of this study was to determine the effect of removal of the smear layer on canal obturation as measured by penetration of bacteria from a coronal direction. Fifty-four extracted human teeth were decoronated and instrumented in a uniform manner. Following instrumentation the root canals of 20 teeth were rinsed with 17% EDTA and 5.25% NaOCl to remove the smear layer before obturation. A second group of 20 teeth were flushed with NaOCl alone. The teeth of both groups were obturated with Thermafil plastic carriers and Roth's sealer. The root canals of another 10 teeth, five rinsed with EDTA and five without, were obturated with Thermafil without sealer. Two teeth serving as positive controls were instrumented but not obturated, while another set of two were sealed coronally and apically to serve as negative controls. The root surface of each tooth was sealed with nail varnish. A small chamber was thoroughly sealed around the coronal aspect of each tooth so that bacteria placed therein could move only through the obturated canal space. Each tooth was placed in a test tube containing sterile trypticase soy broth (TSB). An inoculum of Proteus vulgaris in TSB was placed in each coronal chamber at five day intervals and daily observations were made for bacterial growth in the apical reservoir for 21 days. Both positive control teeth showed bacterial penetration after 24 h. Neither of the two negative control teeth demonstrated penetration for the duration of the study. The frequency of bacterial penetration through teeth obturated with intact smear layer (70%) was-significantly greater than that of teeth from which the smear layer had been removed (30%) P < 0.05. All but one tooth obturated without sealer exhibited bacterial penetration, irrespective of the presence or absence of smear layer. Removal of the smear layer enhanced sealability as evidenced by increased resistance to bacterial penetration. PMID- 9206433 TI - Adherence of Streptococcus gordonii to smeared and nonsmeared dentine. AB - The purpose of this study was to investigate the adherence of Streptococcus gordonii to smeared and nonsmeared dentine and to assess the influence of patent dentinal tubules on bacterial retention. In order to examine bacterial adherence, 10 non-smeared (group 1) and 10 smeared samples (group 2) of outer root dentine were prepared from teeth exhibiting dentine sclerosis. Ten non-smeared samples from inner coronal root dentine that did not exhibit sclerosis were prepared in order to study the influence of patent tubules on bacterial retention (group 3). Cells of the bacterium were radioactively labelled and an adhesion assay was performed. The number of bacteria adhering to the dentine surface was determined by scintillation counting. The results show that the number of bacteria adhering to both smeared and non-smeared outer sclerotic dentine was low (0.3% of inoculum), and there was no significant difference between the groups (P > 0.3). A significantly higher number of bacteria was retained on the inner non-smeared coronal root dentine (P < 0.0001) compared to groups 1 or 2. The results suggest that dentinal smear layers do not enhance or impede bacterial adherence to the dentine matrix. Dentinal surfaces with patent dentinal tubules retain more bacteria than a smeared surface. PMID- 9206434 TI - Assessing apical deformation and transportation following the use of LightSpeed root-canal instruments. AB - The LightSpeed root canal instrument is similar in design to the Canal Master instrument with the exception of being engine driven and fabricated from nickel titanium alloy. The purpose of the design of these instruments is to reduce apical transportation of the canal during cleansing and shaping procedures. Twenty extracted human molar teeth with roots of varying degrees of curvature were used in this study. A double exposure radiographic technique was used to assess the presence or absence of apical transportation resulting between the initial instrument and the final instrument, a size 50 LightSpeed. Only one of the 20 teeth examined exhibited apical transportation. Results of this study suggest that little or no apical canal transportation could be expected when curved canals are instrumented using the LightSpeed root canal instruments. PMID- 9206435 TI - Sterilization of infected root-canal dentine by topical application of a mixture of ciprofloxacin, metronidazole and minocycline in situ. AB - The aim of this study was to observe the potential of a mixture of ciprofloxacin, metronidazole and minocycline to kill bacteria in the deep layers of root canal dentine in situ. After the crowns of extracted teeth had been removed, the drug combination (0.5 mg of each drug), or sterile saline, as the control, was placed in the root canals which had been previously irrigated ultrasonically with G4M EDTA. The penetration and bactericidal efficacy were estimated by various procedures as follows. (1) A cell suspension of E. coli was placed into small cavities prepared parallel to the root canals on the cut planes of nine single rooted teeth. The teeth were then entirely covered with blue inlay wax. At time 0, and at 5h, 24h and 48h after the drug combination had been applied, cells of E. coli were recovered from the cavities by washing the cavities several times with sterile saline solution, and were cultured on the surfaces of heart-infusion (HI) agar plates. Total colony-forming units were tuen counted. Bacterial recoveries decreased with time, and no bacteria were recovered 48 h after application of the drug combination, while bacteria survived in all cases with the controls, (2) After the drug combination or sterile saline had been placed into and sealed in the root canal with blue inlay wax, the teeth were placed into HI agar plates where cells of E. coli had been inoculated. After culturing, a clear zone caused by the inhibition of bacterial growth was observed around the teeth, but not in the control experiment. (3) After sampling infected root dentine of 12 freshly extracted teeth as positive controls, the drug combination (0.5 mg each) was placed in the root canals. No bacteria were recovered from the infected dentine of the root canal wall 24 h after application of the drug combination, except in one case in which a few bacteria were recovered. On the basis of these results, penetration through dentine and antibacterial efficacy of the drug combination can be expected against bacteria infecting the dentine of the root canal wall in situ when the drugs were placed in root canals which had been irrigated ultrasonically. PMID- 9206436 TI - In-vitro antibacterial susceptibility of bacteria taken from infected root dentine to a mixture of ciprofloxacin, metronidazole and minocycline. AB - The aim of this study was to clarify the antibacterial effect of a mixture of ciprofloxacin, metronidazole and minocycline, with and without the addition of rifampicin, on bacteria taken from infected dentine of root canal walls. The efficacy was also determined against bacteria of carious dentine and infected pulps which may the precursory bacteria of infected root dentine. This efficacy was estimated in vitro by measuring bacterial recovery on BHI-blood agar plates in the presence or absence of the drug combination. Bacteria ranging in number from 10(2) to 10(6) occurred in samples of infected root dentine (27 cases). However, none was recovered from the samples in the presence of the drug combination at concentrations of 25 micrograms ml-1 each. The respective drug alone (10, 25, 50 and 75 micrograms ml-1) substantially decreased the bacterial recovery, but could not kill all the bacteria. Bacteria taken from carious dentine (25 cases) and infected pulps (12 cases) were also sensitive to the drug combination. These results may indicate that the bactericidal efficacy of the drug combination is sufficiently potent to eradicate bacteria from the infected dentine of root canals. PMID- 9206437 TI - Periradicular curettage. AB - Periradicular curettage is a part of the treatment procedure of periradicular surgery. Its main purpose is to remove pathological periradicular tissues for visibility and accessibility to facilitate the treatment of the apical root canal system, or sometimes for the removal of harmful foreign materials present in the periradicular area. Inflammatory periradicular lesions (granuloma and cysts) are the responses of the periradicular tissues to irritants from the root canal and not from the periradicular area unless medicaments and/or filling materials have been forced through the apical foramina or perforations into the periodontium. Histologically, the inflammatory periradicular lesion is similar to healing granulation tissue, which is composed of cells which have natural and specific immunological defence capability and cooperate by means of cytokines to amplify the protective mechanisms of the host. Accordingly, it is not necessary to completely curette out all the inflamed periradicular tissues during surgery, since this granulation-like tissue will be incorporated into the new granulation tissue as part of the healing process. To control the source of irritants in the root canal is far more important than to remove all periradicular tissues affected by the irritants. The successful removal of all irritants from the root canal system results in resolution of pulpally induced periradicular lesions. In the case where the periradicular lesion is caused by endodontic instruments or cytotoxic filling materials placed in the periradicular tissues, removal of these foreign objects is required for resolution of the lesion. PMID- 9206438 TI - Pattern of transmission of laser light in teeth. AB - This study examined the transmission of helium neon laser light in 20 dog and human teeth. The effect of probe position and angulation was observed both macroscopically and at a microscopic level using confocal microscopy. In all teeth in both species, laser light was transmitted through teeth to the pulpal surface with the light following the path of the enamel prisms and dentinal tubules. Probe angulation did not affect the pattern of light transmission, nor did probe position; however, the position of the probe on the tooth surface determined which section of the pulp was illuminated. Enamel and dentine together are able to collect and distribute light within the tooth, with both enamel prisms and dentinal tubules acting as optical fibres. PMID- 9206439 TI - Variations in the susceptibilities of components of the endodontic microflora to biomechanical procedures. AB - The role of the endodontic microflora in pulpal disease and in endodontic treatment failures is well established. Thus the need for effective microbial control is one of the important justifications for biomechanical procedures. However the efficacy of this stage of treatment is dependent upon the vulnerability of the involved species, which may not be uniform. The aim of this study was to investigate variations in the susceptibilities of members of the root canal microflora to routine biomechanical procedures. Forty-two root canals were investigated microbiologically. Samples were collected before and after instrumentation and the bacterial findings were compared. In 15 cases of 'primary' root canal therapy, despite changes in the population size, no significant change in the species composition of the microflora was observed. However in 27 cases 'secondary' treatment, a decrease in the number of isolations of Peptostreptococcus spp. was found (P = 0.008). When all 42 cases were considered together, significant decreases were found between first and second samples for anaerobes (P = 0.0117) and for Grampositive species (P = 0.008), especially Peptostreptococcus spp. (P = 0.02). It was therefore concluded that certain species are more resistant to the biomechanical procedures than others. PMID- 9206440 TI - The effect of smear layer on microbial coronal leakage of gutta-percha root fillings. AB - The aim of this in vitro study was to determine the effect of removal of the smear layer on canal obturation as measured by penetration of bacteria from a coronal direction. One hundred and twenty extracted human teeth with straight, single root canals were decoronated. The canals were prepared using the modified double-flared technique with balanced force under copious irrigation. The apical matrix was prepared to size 40 and apical patency subsequently confirmed with a size 15 file. The teeth were divided randomly into experimental groups (80 teeth) and control groups (40 teeth). The root canals of 40 experimental and 20 control teeth were rinsed with 40% citric acid and 2% NaOCl to remove the smear layer before obturation. In experimental groups, 20 teeth with smear layer intact and 20 teeth with smear layer removed were obturated with lateral condensation of cold gutta-percha and Apexit sealer. A further 20 teeth with smear layer intact and 20 teeth with smear layer removed were obturated with the Trifecta technique with the same sealer. In control groups, 10 teeth with smear layer intact and 10 teeth with smear layer removed were obturated with lateral condensation of cold gutta-percha and Apexit sealer. These teeth were completely sealed both coronally and apically to serve as negative controls. The remaining 20 teeth with either smear layer intact or smear layer removed were not obturated and served as the positive controls. The root surface of each tooth was sealed with nail varnish. The cut end of a polypropylene tube was sealed around the coronal part of each root canal so that bacteria placed therein could move only through the obturated canal space. Each root was placed in a glass bottle containing sterile Todd Hewitt Broth (THB) and aliquots of 0.5 ml of THB were injected into the polypropylene tube. The model system was centrifuged at 168 g. An innoculum of Streptococcus sanguis in THB was placed in each coronal chamber at 5-day intervals and daily observations were made for bacterial growth in the apical reservoir for 90 days. All positive control teeth showed bacterial penetration within 24 h, while the negative control teeth remained uncontaminated throughout the test period. Leakage through the experimental teeth was variable ranging from 7 to 86 days. There was no statistical significant difference (P > 0.05) in leakage between the obturated canal when the smear layer was either removed or intact. PMID- 9206441 TI - An in vitro study of the coronal leakage of two root canal sealers using an obligate anaerobe microbial marker. AB - The aim of this in vitro study was to investigate the coronal leakage of obligate anaerobes into root canals obturated with lateral condensation of cold gutta percha with two root canal sealers. Sixty extracted human teeth with straight, single root canals were prepared using the modified double-flared technique with balanced force under copious irrigation until the master apical file was size 40. The teeth were divided randomly into experimental groups (40 teeth) and control groups (20 teeth). In the experimental groups, 20 teeth were obturated with lateral condensation of cold gutta-percha and AH26 sealer and 20 teeth were obturated with the same technique using TubliSeal EWT sealer. In the control groups, 10 teeth were obturated with the same technique either with AH26 or TubliSeal EWT sealer. These teeth were completely sealed to serve as negative controls. The remaining 10 teeth were not obturated and served as positive controls. The root surface of each tooth was sealed with nail varnish except the apical 2 mm. The coronal part of each root canal was sealed with the cut end of polypropylene tube and placed in a glass bottle containing sterile Fastidious Anaerobe Broth (FAB). Aliquots of 0.5 mL of FAB were injected into the polypropylene tube and the model system was centrifuged at 168 g. An inoculum of Fusobacterium nucleatum in FAB was placed in each coronal chamber at 7-day intervals and daily observations were made for bacterial growth in the apical reservoir for 12 weeks. All positive control teeth showed bacterial leakage within a week, while the negative control teeth remained uncontaminated throughout the test period. All the experimental teeth exhibited leakage of bacterial metabolites within 12 weeks, ranging from 1 to 12 weeks. The mean time for complete leakage in the AH26 and the TubliSeal EWT groups was 8.4 and 8.2 weeks respectively. There was no statistically significant difference (P > 0.05) in leakage between the AH26 and the TubliSeal EWT groups. PMID- 9206442 TI - Microleakage of Cavit, CavitW, CavitG and IRM by impedance spectroscopy. AB - The aim of this study was to quantify the sealing ability of four temporary filling materials over 9 days using a new electrochemical technique. Fifty-two extracted human maxillary bicuspids were selected and prepared for the measurements. They were divided into four groups of 12 teeth each, in addition to two positive and two negative controls. After preparation of the endodontic access cavity the sealing ability was registered. After a randomization procedure one group was obturated with IRM, another group with Cavit, a third group with CavitW and the last group with CavitG. The sealability was measured just after obturation (time 0) and after days 1, 2, 3, 4, 7 and 9. The results showed that the IRM group was significantly more watertight than the different Cavit formulations. Throughout the experiment no significant difference was noticed between the Cavit and CavitW groups (P > 0.05). The CavitG group was significantly less watertight throughout the measurements (P < 0.05). PMID- 9206443 TI - Rigidity and retention of carbon fibre versus stainless steel root canal posts. AB - Two of the main requirements of a root canal post are that it is rigid so as to resist flexing under functional load, and that it is well retained in the root. This study compared these properties in two different 1-mm diameter root canal posts--smooth carbon fibre posts (Endopost) and serrated stainless steel posts (Parapost). Ten posts of each type were tested for rigidity in a three-point bend test. Ten posts of each type were cemented with resin cement into the roots of endodontically treated, extracted teeth. The tensile force required to remove the posts was recorded. The Paraposts proved to be significantly more rigid under load (P < 0.001) and significantly more strongly retained in the tooth roots (P < 0.005). The Parapost appears to be a mechanically superior post for the restoration of root-filled teeth with narrow diameter root canals. PMID- 9206444 TI - Resumed tooth development following avulsion of a permanent central incisor. AB - A case is presented in which an upper right central incisor was avulsed because of trauma at age six. At the time of trauma it was suspected that the pulp had remained in the socket although the tooth had been lost. No attempt was made to treat the socket, remove the pulp, or replant the tooth. Follow-up radiography showed that, despite initial apparent complete loss of the calcified portion of the tooth, root development proceeded to completion. PMID- 9206445 TI - Calcium ion diffusion from calcium hydroxide-containing materials in endodontically-treated teeth: an in vitro study. AB - The aim of this in vitro study was to determine the amount and duration of diffusion of calcium ions from both a calcium hydroxide-containing root canal sealer and an intracanal medicament, through the apical foramen and the dentinal tubules of endodontically-treated teeth. The root canals of 88 freshly extracted single-rooted teeth were prepared using the modified double flared, balanced force technique with patency filing. The teeth were divided into four test groups of 20 teeth each and a control group of eight teeth. One group was dressed with a non-setting calcium hydroxide while the other was obturated using cold lateral condensation of gutta-percha with a calcium hydroxide-containing sealer. An artificial root defect was created in the apical third of the root of the teeth in the other two groups and the root canals were either dressed or obturated in the same way as the first two groups. All groups were incubated at 37 degrees C and were sampled for calcium diffusion after 1, 2 and 3 days and 1, 2, 3, 4 and 8 weeks. There was statistically significantly more calcium diffusion with the non setting groups compared with the sealer groups (P < 0.05). There was more calcium ion diffusion from teeth with a patent apical foramen than those with an artificial resorptive root defect (P < 0.05). PMID- 9206447 TI - The application of the buccal object rule for the determination of calcified root canals. AB - Root canal calcification represents a serious problem which often occurs in clinical practice. Calcified root canals are difficult or even impossible to find and treat conservatively. In this paper, a technique is proposed which may be helpful in detecting and locating calcified root canal(s) and properly treating them. The proposed technique is simple and easy to perform in posterior and anterior teeth, and it may be useful in everyday clinical practice. PMID- 9206446 TI - The antimicrobial activity of endodontic sealers to anaerobic bacteria. AB - The effect of five root canal sealers and two root canal dressing materials on the growth of three anaerobic bacteria associated with endodontic infections was determined using the agar diffusion inhibitory test. Samples of the following endodontic sealers (Apexit, Ketac-Endo, Roth Sealer, Sealapex and Tubliseal) and root canal dressing materials (Pulpdent and Root-cal) were incubated for 48 h with the following anaerobic bacteria: Capnocytophaga ochracea, Porphyromonas gingivalis and Peptostreptococcus micros. Statistically significant zones of bacterial growth inhibition for all the bacteria tested were observed in descending order of antimicrobial activity: Roth Sealer, Ketac-Endo, Tubliseal, Apexit and Sealapex. Root-cal and Pulpdent also showed statistically significant antimicrobial activity, but only to Capnocytophaga ochracea, not to the other two bacteria tested, with Pulpdent being the least active. PMID- 9206448 TI - Rethinking the curriculum crunch. PMID- 9206449 TI - Color accuracy of resin cements and try-in pastes. AB - The color accuracy of resin composite cements and their corresponding try-in pastes was assessed. Three shades of three brands were compared using the cements alone and in contact with a standard porcelain disk. There were significant differences in color between resin cements and their corresponding try-in pastes. The addition of a 1-mm-thick porcelain disk greatly reduced the apparent color differences. The color of resin cements changed over time, but not significantly. Baseline measurements of resin cement colors were compared and measurements made after 24 hours of storage. Color changes were statistically significant but not perceptible, and all samples became slightly darker with age. PMID- 9206450 TI - Frictional resistance of removable partial dentures with retrofitted resin composite guide planes. AB - Frictional resistance to the dislodging forces of a removable partial denture with and without resin composite retrofitted guide planes was compared in vitro. Six master models of four natural teeth, each simulating a Kennedy Class III modification 1 arch, were used. Guide planes were prepared, and 18 cobalt chromium removable partial denture frameworks without clasps were fabricated. Three resin composite materials were applied to the guide planes as retrofitting materials. Following this, the resistance to dislodgment of the removable partial denture framework was evaluated under cyclic dislodgment, and the resistance to dislodgment before and after resin composite retrofitted guide planes was compared. The dislodgment force was recorded every 500 cycles. Testing was concluded when the dislodging force value reached 50% of the initial value. There was a significant difference between the dislodging forces with and without retrofitted guide planes. Higher dislodging forces were recorded when resin composite retrofitted guide planes were used. Z100 was the most effective retrofitting material under cyclic dislodgment, and Heliomolar and Herculite XR performed similarly. PMID- 9206451 TI - Retention of ERA direct overdenture attachments before and after fatigue loading. AB - This in vitro study investigated the retention of the four different color-coded ERA attachments prior to and after various levels of fatigue loading (at baseline, at 500 cycles, and after every 500 cycles up to 5,500 cycles). Samples were placed in the Instron testing machine for determination of initial retentive values and then cycled in a specially designed fatigue machine and retested. The results of this study demonstrated that although there are four different retentive elements supplied by the manufacturer of the ERA system, there were only two significantly different groups; (1) the white attachments and (2) the orange, blue, and gray attachments (P < .05). After 500 cycles, there was a loss in retention of 60% for the white, 60% for the orange, 56% for the blue, and 54% for the gray. After 1,500 cycles there was no difference in retentive values for any of the four colored attachments (P < .05). Microscopic examination of worn specimens supported the findings of the fatigue testing. PMID- 9206452 TI - Effect of die spacers on precementation space of complete-coverage restorations. AB - The authors measured the precementation space achieved in vivo with varying layers of die spacer. Two hundred and ninety-one castings were fabricated in the Advanced Education Program in Prosthodontics at New York University College of Dentistry, New York, New York. Each of four clinician groups applied one layer of Taub Die Hardener and from one to four layers of die spacer to within 0.5 to 1.0 mm of the dies' finishing line. The resultant castings were adjusted using GC Fit Checker. After final seating of the casting, the Fit-Checker was removed, sectioned buccolingually, and measured using a micrometer at three locations (midocclusally, midaxially, and on the bevel) for a total of 657 readings. Only after data collection were the specimens identified by group. Analysis of variance demonstrated a significance for group and location. Post hoc one-way analysis of variance and Sheffe tests (P < .1) demonstrated that: (1) one or two layers of die spacer demonstrated smaller precementation spaces than three or four layers; (2) the midocclusal precementation space was greater than the midaxial precementation space; (3) on the bevel area where no die spacer was used, the most consistent and minimal precementation space was measured. PMID- 9206453 TI - Surface layer characterization after dual ion exchange of a leucite-reinforced dental porcelain. AB - The dual ion-exchange technique has been shown to improve the strength of feldspathic dental porcelains by first replacing constituent alkali ions with smaller ions above the strain point, and then exchanging the smaller ions for larger ions at temperatures below the strain point. The strength increase is directly related to the thickness of the surface-exchanged layer. This study evaluated the thickness of the exchanged layer after dual ion exchange of a leucite-reinforced dental porcelain. Optec HSP porcelain disks were fabricated, submitted to a dual ion-exchange treatment, and indented at various loads before determining the biaxial flexure strength. The mean flexural strength for the ion exchanged groups was significantly higher than for control groups, except when the depth of the median crack exceeded 138 microns. Wavelength Dispersive Spectrometry analyses on cross sections showed that the mean potassium amount in the glassy matrix was significantly lower (P < .001) for the treated specimens than for the controls. The mean thickness of the exchanged layer after dual ion exchange was estimated at 140 microns. PMID- 9206454 TI - Impact strength of a modified continuous glass fiber--poly(methyl methacrylate). AB - The effect of fiber reinforcement of autopolymerizing poly(methyl methacrylate) was investigated. The impact strength of continuous E-glass fiber-poly(methyl methacrylate) composite was determined. Rectangular test specimens (n = 10 per group) were modified by incorporating an additional fiber reinforcement of untreated E-glass fibers, silanized E-glass fibers, or aramid fibers in the test specimens. Controls were either unreinforced or reinforced from the middle of the test specimen only. The impact strength of the specimens was measured by using a charpy-type pendulum impact tester after the specimens had been stored in water at 37 degrees C for 4 weeks. After the impact strength test, the length of the delamination of poly(methyl methacrylate) from the fibers was measured and plotted to the impact strength of the test specimens by using a linear regression model. The impact strength of unreinforced autopolymerizing poly(methyl methacrylate) was 7.8 kl/m2, while incorporation of glass fiber reinforcement with a fiber concentration of 12.4 wt% increased the impact strength to 74.7 kl/m2 (P = .000). The additional fiber reinforcement of the test specimen did not affect the impact strength (P = .363). Delamination negatively correlated with the impact strength of the test specimens (r = -.72, P = .000). The results of this study suggest that glass fiber reinforcement enhanced the impact strength of autopolymerizing poly(methyl methacrylate), while the use of additional fiber reinforcement made of aramid or glass fibers in the test specimens did not have an effect on the impact strength. PMID- 9206455 TI - Ten-year follow-up study of conical crown-retained dentures. AB - This study reports the results of a long-term evaluation of conical crown retained dentures. One hundred fifty-two restorations that had been in place for over 10 years were analyzed. Restorations were divided according to the Kennedy classification. Abutment and periodontal health, occlusion, retention, and frequency of relining and repair were among the factors evaluated. Good prognoses of removable partial dentures were shown in Kennedy Classes I, II, and III arches. However, some of the restorations placed on only a few remaining abutments showed rather unfavorable situations with almost all of the factors evaluated. PMID- 9206456 TI - The effect of alveolar bone grafting on the prosthodontic/reconstructive treatment of patients with unilateral complete cleft lip and palate. AB - This study compared the effect that the introduction of mixed dentition alveolar bone grafting and subsequent orthodontic treatment has had on the prosthodontic/reconstructive habilitation of patients with unilateral cleft lip and palate. Two groups, each consisting of 40 consecutive patients with unilateral cleft lip and palate, were compared at the end of their dental treatment. In the group treated prior to the advent of bone grafting, all subjects received a fixed partial prosthesis in the cleft area, and a total of 87 abutment teeth were prepared for complete coverage crowns. In contrast, in the group of patients for whom the alveolus was restored by bone grafting, it was possible to obtain a complete dental arch without prosthodontic intervention in 36 patients (90%). Thirteen subjects in the bone grafting group had the crown anatomy of anterior teeth modified using resin composite or resin-bonded porcelain veneers. Two patients had a premolar transplanted to the anterior region of the dental arch. On average the dental treatment was completed 3 years earlier in the bone grafting group. Thus, alveolar bone grafting with subsequent orthodontic treatment, together with advances in dental materials, have markedly reduced the need for prosthodontic procedures and have also allowed the completion of the dental treatment at an earlier age. PMID- 9206457 TI - The in vitro color stability of acrylic resins for provisional restorations. AB - This study made an in vitro comparative evaluation of the color variation of four types of acrylic resin for provisional fixed prostheses, as determined using computerized spectrophotometry, before and after a 20- and 30-day cycle of immersion in four staining solutions. The four acrylic resins used for provisional fixed restorations were: Cold Pac, Trim, Protemp, and Mixacryl II. Thirty-two specimens for each resin were divided into four subgroups of eight elements, immersed in the four staining solutions (synthetic saliva, synthetic saliva and tea, synthetic saliva and coffee, and synthetic saliva and chlorhexidine in 0.12% water solution), and then placed into four thermostatic baths at 37 degrees C +/- 1 degree C. All specimens were measured for each resin before immersion (baseline). After 20 and 30 days, the specimens were analyzed by computerized spectrophotometry and compared. All data were analyzed with analysis of variance for repeated measures (P < .05). Only the Cold Pac resin was color stable in all staining solutions, while the others showed color changes from the different staining solutions. PMID- 9206458 TI - The influence of different cementation modes on the fracture resistance of feldspathic ceramic crowns. AB - One hundred twenty pressed feldspathic ceramic crowns were luted to 20 steel dies using six different cementation modes. Fracture resistance was tested under an angle of 45 degrees and was determined as the maximal fracture load. Crowns were tested with luting agent only (groups A and C) and after etching with hydrofluoric acid, silanating, and the application of a bonding agent (groups B, D, E, and F). The resulting means were: phosphate cement 294.3 (A) and 282.2 (B); glass-ionomer cement 217.2 (C) and 255.4 (D); resin composite 382.2 (E) and 687.6 (F). Statistical analysis revealed significantly greater fracture resistance (P < .01) of resin luted crowns. Bonding to the die almost doubled the fracture resistance. Conditioning of the inner surfaces of the crowns did not improve the fracture resistance of crowns luted using zinc phosphate or glass-ionomer cements. PMID- 9206459 TI - Bond strength of six soft denture liners processed against polymerized and unpolymerized poly(methyl methacrylate). AB - The bond strength of six commercial soft denture liners was evaluated by a two phase tensile test. The soft denture liners investigated were VinaSoft, Prolastic, Flexor, Molloplast-B, Novus, and SuperSoft. The samples were fabricated by processing them (1) against polymerized poly(methyl methacrylate), and (2) against unpolymerized poly(methyl methacrylate). The soft denture liners were processed according to the manufacturers recommendations. The samples were tested using an Instron Universal Testing Machine. The mode of failure, adhesive or cohesive, was also recorded. The bond strength when processed against unpolymerized poly(methyl methacrylate) ranged from 0.48 to 2.60 MPa, and when processed against polymerized poly(methyl methacrylate) the bond strength ranged from 0.94 to 2.56 MPa. A two-way analysis of variance (P = .05) revealed a significant increase in bond strength when the liners were processed against polymerized poly(methyl methacrylate), except for Novus, which had no change, and VinaSoft, which decreased. The Tukey interval between materials was .22 and between methods of polymerization was .08. Four of the six liners investigated demonstrated increased bond strength when processed against polymerized poly(methyl methacrylate). It was concluded that bonding can be influenced by the processing method. PMID- 9206460 TI - Radiographic findings, ridge resorption, and subjective complaints of complete denture patients. AB - To evaluate the relationship between the complaints of complete denture wearers and alveolar bone resorption as well as the location of mental foramina, 96 patients were interviewed. All subjects had received new dentures at the University of Iowa between August 1985 and July 1990. Panoramic radiographs had been made for all the subjects before dentures were fabricated. The amount of estimated ridge resorption correlated significantly with the number of years females had been edentulous, but no correlation was found in males. The location of the mental foramen in relationship to the crest of residual ridge correlated negatively with the number of years both genders were edentulous. Subjective need for dental treatment, as expressed by "sore gums" or poor or fair chewing ability, were the most frequent complaints among the subjects. These complaints were more often recorded with the subjects who had lost more than 50% of their estimated original ridge height than with those with less than 50% resorption, but this difference was not significant. PMID- 9206461 TI - Amputation neuroma following radical neck dissection--report of 3 cases. AB - Amputation neuroma occurred in three cases among 111 cases of oral cancer patients after radical neck dissection. All of three cases appeared as a small nodule at superior neck around the carotid artery with accompanying tenderness. The lesions were located in continuity with the proximal end of nerve at excision. Although the incidence of amputation neuroma is low, critical examination is required to distinguish this from recurrent tumor after cancer surgery. PMID- 9206462 TI - Antibacterial effect of composite incorporating Triclosan against Streptococcus mutans. AB - It has been shown that composite incorporating the antibacterial agent Triclosan (Irgasan DP 300, which is sparingly soluble in water, inhibited in vitro plaque formation by Streptococcus mutans, although the release of the agent was much less than the minimum inhibitory concentration for the bacterium. In this study, the inhibitory effect of the composite incorporating 1% Triclosan against growth and adherence of S. mutans was investigated. S. mutans was inoculated on the surface of a specimen made of control or Triclosan-incorporated composite and the number of bacteria was compared after 3, 6, 12 and 24 hr of incubation. The adherence of S. mutans to the control and experimental composite, with or without saliva treatment, was also examined by scanning electron microscopy. The composite incorporating Triclosan demonstrated significant inhibition of growth of S. mutans after 6, 12 and 24 hr of incubation. Adherence of S. mutans to the Triclosan-incorporated composite was less compared with control for both non treated and saliva-treated specimens. It is concluded that the antiplaque effect of composite incorporating Triclosan depends upon its ability to inhibit bacterial growth and adherence, and Triclosan-incorporated composite is able to exhibit the antibacterial activity even after being treated with saliva. PMID- 9206463 TI - Malignant transformation of leukoplakia three times in period of 11-years--case report. AB - Leukoplakia is a commonly occurring precancerous lesion. The following case report describes a patient who had multiple leukoplakia in this mouth. He had three sites of leukoplakia, the left hard palate, the gum of the right maxilla and the gum of the left mandible, all of which underwent malignant transformation during the period of 11-years after diagnosis. PMID- 9206464 TI - Effects of salvia miltiorrhiza bunge (SMB) on MC3T3-E1 cells. AB - In previous research, we found that the Salvia Miltiorrhiza Bunge (SMB), a traditional Chinese medicinal herb, accelerates orthodontic tooth movement. In the present study, to characterize the actions of SMB on bone remodeling, we investigated the effect of SMB on DNA synthesis and alkaline phosphatase activity of murine osteoblast like cell-clone, MC3T3-E1 cells in vitro. Treatment of the cells with SMB for 72 hours caused significant increase in ALPase activity. It was found that SMB increased ALPase activity in a dose-dependent manner, and up to maximum at the concentration of 5.0 mg/ml. At that concentration, ALPase activity was about 135% greater than that of control. SMB at 5.0 mg/ml significantly stimulated ALPase activity of the cells in multilayer on day 8 or calcification stage on day 16, but inhibited it in the sparse stage on day 2 or subconfluency stage on day 4. SMB had no effect on DNA synthesis in any stage of culture. ALPase activity significantly increased at 48 hours and was up to 300% greater than that of control activity at 96 hours. These findings suggest that SMB directly stimulates ALPase activity of MC3T3-E1 cells in multilayer stage or calcification stage without any effect on proliferation. Locally administrated SMB may affect the differentiation of osteoblasts in vivo. PMID- 9206465 TI - The incidence of arrhythmias during induction of general anesthesia. AB - In this study, our induction methods of endotracheal anesthesia was evaluated with reference to electrocardiogram, hemodynamic status and arterial blood gas analysis on 153 patients. From the beginning of induction, electrocardiogram was recorded continuously to the completion of intubation. The blood pressure and heart rate were also measured. Arterial blood samples were taken at 40 seconds of apneic period after the mask was removed from the patient's face (CONT group), at 40 seconds of apnea with intratracheal spray (LIDO group), and after intubation following intratracheal spray (INT group). In the electrocardiographic survey, arrhythmias during intubation were rare (1/113, 0.9%). PaCO2 values in INT group showed a significant elevation (+ 14.0 mmHg; p < 0.01) compared to these in CONT group. As a result of careful and gentle induction techniques, the incidence of arrhythmias during intubation was very low. However, an elevation of PaCO2 was not avoidable, even in smooth and successful intubation. In conclusion, the importance of more adequate ventilation coupled with skillful intubation in a shorter period to avoid hypercapnia and arrhythmias is appreciated again in the endotracheal anesthesia for the maxillofacial surgical patients with anatomical airway problems. PMID- 9206466 TI - Hardness of denture reline materials polymerized with different techniques. AB - Two kinds of hard type self-cured denture reline materials were polymerized using three different techniques: (1) on a surface of glass plate, (2) under water, (3) under water with some hardening agent. Hardness of specimens were measured longitudinally during three weeks periods of storage in water at 37 degrees C. Results of our study indicated that the specimen cured with the special hardening agent showed the higher hardness than those cured only in water. Most of the hardness changes took place during first three days of the experiment period. PMID- 9206467 TI - A method of three-dimensional measurement and evaluation of external nasal forms. AB - An accurate, quantitative method of measuring external nasal forms was described to evaluate the results of rhinoplasty, in which facial plaster models were measured with a highly accurate contact-type three-dimensional coordinate measurement apparatus. Furthermore, two original programs to identify facial landmarks objectively and to measure the curved surface of the external nasal form in order to evaluate cleft lip nasal deformities were described. These provided accurate and objective data for the facial landmarks and the curvature of nasal alae. PMID- 9206468 TI - Additional heat-curing of light-cured composite resin for inlay restoration. AB - The purpose of this study was to evaluate the effect of heat application after light curing on some physical properties of composite resin for dental inlay. Specimens each were additionally dry heat-cured at various temperatures for 15 min after initial light curing, and other specimens were not heat-cured as a control. Fracture toughness, bending strength, bending elastic modulus, coefficient of thermal expansion, hygroscopic expansion, microhardness, water absorption and solubility were determined. Fracture toughness, bending strength and bending elastic modulus were significantly higher when heated at 80-120 degrees C than the control. The coefficient of thermal expansion, hygroscopic expansion, microhardness and solubility were significantly improved than those of the control when heated, while water absorption was not significantly altered by dry heat-curing. These results indicate that the physical properties of composite resin, except water absorption, were improved significantly when heated after initial cure. PMID- 9206469 TI - Guidelines for managing the orthodontic-restorative patient. AB - Occasionally, patients require restorative treatment during or after orthodontic therapy. Patients with worn or abraded teeth, peg-shaped lateral incisors, fractured teeth, multiple edentulous spaces, or other restorative needs may require tooth positioning that is slightly different from a nonrestored, nonabraded, completely dentulous adolescent. Generally, orthodontists are not accustomed to dealing with patients who require restorative intervention. Should the objectives of orthodontic treatment differ for the restorative patient compared with the nonrestorative patient? How should the teeth be positioned during orthodontic therapy to facilitate specific restorations? Should teeth be restored before, during, or perhaps after orthodontics? The answers to these and other important questions are vital to the successful treatment of some orthodontic patients. This article will provide a series of eight guidelines to help the interdisciplinary team manage treatment for the orthodontic-restorative patient. PMID- 9206470 TI - Managing treatment for the orthodontic patient with periodontal problems. AB - Some adult patients have mild to moderate periodontal disease before orthodontic treatment. These patients may be at risk of developing further periodontal breakdown during orthodontic therapy. However, careful diagnosis and judicious management of these potentially volatile patients can alleviate the risk. In this article, the diagnosis and management of several periodontal problems is discussed. The need for and timing of preorthodontic periodontal surgery for these situations is elucidated. In addition, the types of tooth movement that will ameliorate these problematic situations is described. This information is valuable for the orthodontist who treats patients with underlying periodontal problems. PMID- 9206471 TI - Orthodontic-endodontic treatment planning of traumatized teeth. AB - Occasionally, an orthodontic patient will accidentally traumatize a maxillary anterior tooth before or during orthodontic treatment. In some situations, the trauma will be substantial and avulse the tooth. In other accidents, the tooth may not avulse, but the pulp becomes nonvital. If the pulp is devitalized, and the root has not fully formed, the apex of the root canal may be wide. In this situation, the endodontist may recommend apexification procedures to help close the apex before conventional obturation of the root canal. If the patient is currently undergoing orthodontic movement of the traumatized incisor, what effect will the tooth movement have on the success of the apexification? If the tooth were avulsed, replanted, and then ankylosed, should it be extracted? If so, when should the ankylosed incisor be removed? What effect will further facial growth have on the ankylosed tooth and the potential to achieve a successful esthetic restoration? The answers to these questions are important during the interdisciplinary treatment planning of the patient with traumatized teeth. This article will elucidate the endodontic-orthodontic considerations for patients with traumatized anterior teeth. PMID- 9206472 TI - Interdisciplinary management of single-tooth implants. AB - Orthodontists treat many patients who are missing maxillary lateral incisors and/or mandibular second premolars. In the past, if the canines could not be substituted for lateral incisors, conventional full-coverage bridges were the common restoration. Recently, resin-bonded Maryland bridges became a popular substitute for conventional bridges to avoid crowns on the nonrestored abutments. However, resin-bonded bridges have a poor long-term prognosis for retention, lasting on average about 10 years. Since implants were introduced into dentistry by Swedish researchers in the mid-1980s, they have become a promising substitute for conventional or resin-bonded bridges. However, to successfully place and restore single-tooth implants in young orthodontic patients several questions must be answered. This article will discuss the many interdisciplinary issues that are involved in placing and restoring single-tooth implants in orthodontic patients. PMID- 9206473 TI - Editorial comment on hemodynamic evaluation of the CarboMedics R, St Jude Medical HP and Sorin-Bicarbon valve in patients with small aortic anulus: Noera et al. PMID- 9206474 TI - [Intestinal invagination in childhood: etiopathogenetic evaluations and details of surgical technique]. AB - Intussusception in childhood, especially the idiopathic type, is herein considered. The Authors evaluate some etiopathological aspects on the basis of their experience in 34 patients, and affirm the need of an early and correct diagnosis. They also consider therapeutical management and surgical approach, evaluating local and general conditions of the patient as well as time passed since the onset of symptoms. Finally the importance of an accurate diagnostical examination during the quiescent period and some details of the surgical technique are stressed. PMID- 9206475 TI - [Hypoxemia and pulmonary atelectasis after laparoscopic cholecystectomy]. AB - The aim of this study is to evaluate, after laparoscopic (CL) or open (CO) cholecystectomy, the incidence of pulmonary atelectasis and hypoxaemia which are strictly related to the onset of pulmonary complications. Two groups of 20 consecutive patients affected by symptomatic and not complicated gallstone disease were cholecystectomized either using CO or CL. Hypoxaemia was assessed preoperatively and after operation. Postoperative determination was performed at the 4th, 8th, 12th and 24th hour and then every 12 hours until discharge from hospital. A not informed radiologist evaluated atelectasis through two X-rays, preoperatively and postoperatively at the 3rd day. Atelectasis cases were divided in micro, focal, segmental, lobar and of the entire lung. Statistic analysis was performed using the "t" Student test. No mortality or intraoperative complications occurred. The two groups were similar for age, sex, smoker percentage, obesity, preexisting pulmonary dysfunctions and anaesthesiological risk (ASA). Operative time resulted in longer in CL compared to CO patients although in an insignificant way. PO2 value resulted significatively reduced (P < 0.05) at 4th, 8th, 12th and 24th postoperative hour after CO, while subsequent measurements did not show significant differences. There was no evidence of PO2 significative reduction after CL. After operation atelectasis was found in 11 patients (55%) of CL group (P < 0.05) and in 17 patients (85%) of CO group (P < 0.001). Atelectasis observed in the group of 11 CL patients was represented by 7 micro and 4 focal types, while in the CO group 7 micro, 8 focal and 2 segmental types were found. This study suggests that CL alters the pulmonary function less than CO. PMID- 9206476 TI - [Treatment of peripheral arterial embolisms. Our case series and review of the literature]. AB - Acute arterial obstruction of the limbs represents one of the most frequent events in Emergency Surgery. In 95% of the cases, the cause has to be searched in embolus parting from the heart in patients with rheumatic or fibrillary disease. Currently the two therapeutic methods used for peripheral arterial obstruction are thrombolytic therapy and surgical dysobstruction using Fogarty's catheter. The Authors compare the two methods on the basis of their experience in 129 cases underlying how the thrombolytic therapy (Urokinase, Streptokinase) should be instituted in the early hours from presentation of symptoms, otherwise, the possibility of revascularization will heavily drop. Better results are obtained by positioning a catheter under radiologic guide for intra-arterial infusion. The Authors also believe that up-to-date the surgical approach with Fogarty's catheter represents one of the best procedures, either for its feasibility of for the costs of the thrombolytic therapy. Furthermore, the thrombolysis may be not complete, and account for possible haemorrhagic complications. PMID- 9206477 TI - [Hand ischemia due to "steal syndrome" in vascular access for hemodialysis]. AB - Among complications of vascular access operations, symptomatic steal syndrome is uncommon, but may lead to ischemia of the hand. Between 1983 and 1995, 5 patients with hemodialysis fistulas presented rest pain of finger necrosis with a wrist brachial index of 0.56 (range 0.35 to 0.63) improving to 0.96 (range 0.72 to 1.05) after digital pressure of the fistula. Ligation of distal radial artery was performed in 3 patients with side-to-end radiocephalic fistula, while basilic vein was distally ligated in a case of side-to-side brachiobasilic fistula. A vein "banding" procedure reduce fistula flow and improved distal perfusion in one patient, while a true venous aneurysm of the cephalic vein was treated by excision and replacement with a tapered PTFE graft. Hemodynamic assessment is required during surgical correction, but it may also be useful in pre- and intra operative evaluation of patients undergoing therapeutical AVFs to prevent arterial insufficiency of the hand. PMID- 9206478 TI - [Ulcerated carcinoma of the breast in an elderly woman. An unusual clinical case report]. AB - The authors analyze the most current methods for the treatment of ulcerated breast cancer in the elderly. They describe a peculiar case recently observed, characterized by an ulcerative lesion spreading from the hemiclavicle to the foreaxilla including the axillary cavity, causing a large phlebo-lymphaedema, anaesthesia and paresis of the homoteral upper limb. After an initial treatment with tamoxifen a scapulo-humeral disarticulation and a suture of the cutaneous wide defect were performed using a myo-skin graft which included the deltoid muscle. A radiotherapeutic treatment followed by tamoxifen therapy was carried out. No distant metastases and no local relapses were registered in the one year follow up. The Authors on the basis of their experience and according to the review of the Literature suggest that age itself is not a limiting factor to the therapeutic approach of ulcerated breast cancer. PMID- 9206479 TI - [Surgical strategy in gastric cancer in aged patients]. AB - The trend of the National mortality rate for gastric cancer in the general population and in the old people between 1955 and 1990 was studied. A decreased incidence of mortality in the general population whereas an increased mortality rate in elderly was found. Between 1990 and 1994, 17 patients over 70 years of age were operated on for gastric cancer. The influence of the ASA classification on postoperative complications was then evaluated registering a significant statistical difference between patients included in ASA III group and or > and major complications and mortality (P = 0.044). The surgical strategy in the management of elderly patients with gastric cancer is also discussed. PMID- 9206480 TI - [Nodular fasciitis: a case arising from the splenius cervicis muscle]. AB - An unusual case of nodular fasciitis, arising from the splenius muscle, is presented. This uncommon lesion is always benign, but a local spread in the surrounding muscular tissue is possible. Through a Literature review, the main pathological, clinical and therapeutic features are discussed. PMID- 9206481 TI - [Reconstruction of the defect in the parotid region]. AB - The authors report their experience in the surgical management of a patient with an ulcerated tumor of the neck extending to the parotid region. The different types of reconstruction after radical surgery as well as the choice and reasons that have suggested the use of pectoralis major myocutaneous flap are discussed. PMID- 9206482 TI - [Role of ultrasonography in the study of patients with acute pancreatitis]. AB - The aim of the present study is to establish the role of ultrasound examination (US) in the diagnosis of acute pancreatitis and its ability in differentiating mild or edematous acute pancreatitis (EAP) from severe or necrotizing acute pancreatitis (NAP) in order to indicate further diagnostic procedures and appropriate therapy. Forty patients with clinical suspect of acute pancreatitis were examined with US. All of them were followed-up clinically and with laboratory tests. Nineteen patients underwent a CT examination with contrast media and one without. In 28 cases US diagnosed acute pancreatitis in agreement with clinical and laboratory data. US identified 19 patients with EAP and 9 with NAP. CT, performed in 20 cases on the basis of clinical and US findings, confirmed US diagnosis in 19 cases while in 1 patient CT identified a NAP instead of the EAP suggested by US. According to our results, US is an easy-to-perform and accurate method for the diagnosis of acute pancreatitis and effective in differentiating EAP from NAP; it is also useful in suggesting further diagnostic procedures such as contrast-enhanced CT required in patients with NAP to assess the extension of peripancreatic fluid-collections and possible complications. PMID- 9206483 TI - [Pneumatic dilatation in the treatment of esophageal achalasia]. AB - The purpose of therapy in esophageal achalasia is to reduce the pressure at the level of the lower esophageal sphincter. In this study 26 patients (16 males and 10 females) between 30 and 50 years of age, affected by esophageal achalasia underwent esophageal dilatation with Rigiflex pneumatic dilators. A total of 40 dilatations were performed. A complete success using the Rigiflex pneumatic dilators, was achieved in 24 out of 26 patients (92.3%). PMID- 9206484 TI - [Retroperitoneal sarcomas: recent advances in nosographic framework, diagnosis and integrated surgical treatment]. AB - In this paper the Authors reviewed the recent literature for a more comprehensive and clear vision of the epidemiological and pathological aspects of retroperitoneal sarcomas. The most effective procedures for a an early and accurate diagnosis were identified. Moreover, the different therapeutic choices were taken into account focusing on those provided of the major potential in terms of oncologically valid treatment. PMID- 9206485 TI - [Iatrogenic biliary lesions and stenosis]. AB - Pathogenetic, diagnostic and therapeutic aspects of postoperative bile duct injuries are reviewed. Treatment options are discussed in relation to the time of diagnosis. Lesions detected during the same operation must be immediately repaired through an end-to-end biliary anastomosis or a bilioenteric anastomosis. In limited lesions of the bile duct a T-tube placement should be sufficient. Bile duct lesions recognized postoperatively can be managed through a multimodal surgical, endoscopic, and radiologic approach. In the early postoperative period, surgery is indicated when a complete section of the biliary tract or a severe peritonitis is recognized, or when endoscopic and radiologic treatment has failed. Surgery is also the treatment of choice in the late complete stenosis of the bile duct. Roux-en-Y hepatico-jejunostomy is the most common surgical procedure for the treatment of bile duct lesions and strictures. However, in high bile duct lesions, especially if the risk of anastomotic dehiscence is increased the Authors emphasize the Rodney-Smith technique for the reconstruction of the biliary tract. PMID- 9206486 TI - [Controversies in timing of preoperative staging of pancreatic carcinomas]. AB - In recent years pancreatic cancer has shown an increasing incidence. Preoperative staging represents a main problem for its surgical management. Recent behaviour in the treatment of the disease led to considerably more encouraging results. The Authors describe the cases treated at the Dept. of Surgery of the University of Perugia and review the most important international reports on preoperative staging of pancreatic cancer. PMID- 9206487 TI - [Lacking effect of timing of surgery in relation to the menstrual cycle on prognosis of breast cancer in premenopausal women]. AB - The influence of timing of surgery in relation to menstrual period on survival of breast cancer patients has been both advanced advocated and disputed. A meta analysis on published series showed a statistically significant overall odds reduction when surgery is performed in the luteal phase. The records of 165 premenopausal M- breast cancer women, not on hormonal therapies, consecutively operated on from 1977 to 1991 were reviewed. All patients underwent modified radical mastectomies or quadrantectomies plus operative radiotherapy, Node positive patients received standard adjuvant chemotherapy. Cox regression analysis was used to estimate the relative risk (RR) of death in three models including timing of surgery, age, histology, pathological T and N. In each model, patients were divided into two groups according to the criteria proposed by Badwe, Hrushesky, and Senie. Multivariate analysis showed a significant association between pT and pN and survival, whereas no association with survival was observed for timing of surgery according to Badwe or Hrushesky or Senie criteria (RR = 1.26, RR = 0.91, and RR = 0.88 respectively). Up-to-date agreement on the menstrual phase and relative expected better prognosis is still lacking. PMID- 9206488 TI - [Warthin's tumor of the parotid gland]. AB - The Authors report 6 cases of Warthin's tumor of the parotid gland recently observed. In all cases the tumor was intraglandular, single, and no noteworthy disorders or symptoms except for a 2 to 5 cms tumefaction were present. At surgery all lesions appeared well-capsulated and the exeresis followed a cleavage plane easily identifiable which enabled the preservation of the residual glandular tissue as well as the surrounding nervous and vascular structures. All the patients, up to now, are free from relapse. PMID- 9206489 TI - [Clinico-diagnostic aspects and therapeutic considerations of the combined surgical and radiotherapeutic approach in 2 cases of sacrococcygeal chordoma: our experience]. AB - Chordomas are rare tumours arising from embryonic notochord tissue remnants. The commonest affected segment is the sacrum. This localization may present diagnostic and therapeutic problems. Two cases of chordoma treated by surgery and radiation are reported. PMID- 9206490 TI - [Role of laparoscopy in abdominal traumas: a case report]. AB - The Authors report a case of abdominal traumatism by gunshot wound, presenting with retroperitoneal hematoma and hematoma of the mesocolon, treated in emergency laparoscopy. After a literature review and their case report analysis, they confirm the importance of emergency laparoscopy in hemodynamically stable patients, as well as the diagnostic value of peritoneal lavage, positive for minimal and moderate hemoperitoneum. Laparoscopy allows, as the same time in most cases, the diagnosis and the management of the lesion, while surgery is reserved to the cases of severe hemoperitoneum and perforating injuries. The use of laparoscopy in abdominal blunt injuries, has reduced the number of unnecessary laparotomies with related higher incidence of morbidity. The Authors conclude confirming the importance of this technique for its diagnostic efficacy, lower costs, minimal traumatism, and good aesthetic results as observed in the case report for the modality of the injuries, the young age and the previous sternotomy of the patient. PMID- 9206491 TI - [Intestinal invagination in adults: 3 case reports]. AB - The Authors report their experience in the surgical management of three adults affected by intussusception. The different symptoms and different diagnostic approach, compared to those of childhood, induced the Authors to some evaluations on pathophysiology of intussusception in adults. On the basis of the different pathogenesis some differential criteria between the two forms are stressed, finally suggesting a systematic surgical approach in adults. PMID- 9206492 TI - [Esophageal leiomyoma: ultrasonographic endoscopic diagnosis]. AB - Leiomyoma is the most frequent benign neoplasia of the esophagus. It is generally diagnosed, accidentally during a radiologic examination (filling defect with clear and regular margins) or endoscopically (sessile, hemispheric, covered by pink mucosa). Recently, to the above conventional exams, endoscopic ultrasonography has been added allowing to identify the single layers of the esophageal wall, thus furnishing useful informations on the morpho-structural characteristics of the leiomyoma. From October '94 to May '96, at the Endoscopy Service of the Institute of Oncology of the University of Messina, 12 patients, 8 males and 4 females, ranging from 39 to 69 years of age (median age 55.4) underwent EUS for suspect leiomyoma. An Olympus EU-M20 echoendoscope was used with a radial scan transducer of 12 Mhz. In 8 patients the leiomyoma was located in the III mid-esophagus, while in 4 patients the III inferior portion was interested. The Authors observed lesions ranging in size from 0.5 to 2.5 cm. In their experience, a suspect of leiomyoma represents a good indication for an endoscopic ultrasonography, which shows high sensitivity and specificity. PMID- 9206493 TI - [Mechanical endarterectomy: review of the literature and description of an original device]. AB - The Authors, after a Literature review on endovascular invasive procedures used for inferior limbs obstructive arteriopathy, describe the use of a new device for mechanical endarterectomy. Through the latest acquired experience the importance of dissecting progressively the single layers of the atheroma plaque without arriving to a complete denudation of the arterial wall, so avoiding the risk of myointimal hyperplasia reactions, is outlined. The possibility of using endoarterial stents in case of more indaginous recanalization is also stressed. PMID- 9206494 TI - [Performance and complications of totally implantable port device in bolus hepatic intra-arterial chemotherapy]. AB - The performances of totally implantable ports were analyzed in patients with colorectal metastases undergoing intraarterial treatment. Seventy-nine patients received bolus infusion of Cisplatin (DDP, 57 cases) or Epirubicin (EPI, 22 cases) every 21 and 7 days, respectively. Disease progression or toxicity were the most common causes of interruption of treatment, whereas failure of ports occurred in six and two patients out of DDP and EPI groups, respectively. The incidence of single problems for each port was 65% in DDP group and 64% in EPI group, whereas rate of complications for each patient was 30% and 32%, respectively. The 12-months device duration rate in the two groups was 65% (median 17 months) in DDP group and 78% (median 18 months) in EPI group. The implantable ports employed for bolus arterial infusion, allowed adequate treatment periods in most cases, without any difference as far as intervals between cycles is concerned. PMID- 9206495 TI - [Surgical treatment of spontaneous pneumothorax: comparison of thoracotomy and thoracoscopy]. AB - The video-thoracoscopic treatment of spontaneous pneumothorax currently has the same role of laparoscopic cholecystectomy in abdominal surgery. The Authors consider thoracoscopic approach and traditional thoracotomy examining advantages versus disadvantages, comparing 50 patients with spontaneous pneumothorax treated by thoracoscopy, from February 1992 up to February 1995, and 50 patients, previously treated by open surgery. Video-thoracoscopy has the same percentage of recurrences of thoracotomic approach but assures a quicker functional recovery and, above all, a remarkable reduction of pain. PMID- 9206496 TI - [Use of endoprosthesis in the treatment of airway pathology]. AB - In patients affected by benign or malignant inoperable airway obstructions, therapeutical options include endoscopic treatment by Nd-YAG laser therapy, tracheobronchial dilatations with rigid or flexible bronchoscope, and inflating balloon dilators. Metal self-expanding or silicone stents allow to obtain stable results. Our experience is based on the use of 14 stents (10 Dumon and 4 Wallstent) in 13 cases of stenosis either due to vegetating malignant tumours not amenable to surgery or benign stenosis. In the following 24-48 hours both subjective and objective changes of the pulmonary function, blood gas analysis and radiologic aspects were observed. The results showed an improvement in the respiratory parameters and a sensible improvement in the quality of life. PMID- 9206497 TI - [Use of Doppler color ultrasonography in the microsurgical treatment of idiopathic varicocele]. AB - The aim of this paper is to evaluate the role of Doppler and echo color-Doppler in the microsurgical treatment of varicocele. Since December 1993, 87 consecutive patients underwent microsurgical treatment of varicocele. The study demonstrates that Doppler and Color-flow-Duplex scanning provide an accurate, non-invasive method to identify the position of accessory spermatic veins, evaluating at the same time the flow characteristics of the microsurgical anastomosis. PMID- 9206498 TI - [Unusual pathology of the spleen]. AB - The Authors report 7 cases of unusual spleen pathology represented by 2 real cysts, 1 pseudocyst, 1 hydatid cyst, 2 abscesses, 1 splenic artery aneurysm. Pathophysiological hypotheses are examined, and clinical and diagnostic considerations are stressed, evaluating particularly anatomo-histologic characteristics and the differential diagnosis through non invasive imaging techniques. All patients were subjected to splenectomy via laparotomy, except for the case of real cyst where an inferior hemisplenectomy was performed. In the case of splenic artery aneurysm, an urgent aneurysmectomy was required together with splenectomy, considering the young age of the patient (29 years old), and the type of aneurysm presenting in a double rupture phase. Reoperation was necessary in only one patient to drain an hematic abscess. After a 9 year follow up all the patients are well. PMID- 9206499 TI - [Current trends in therapeutic indications in gallstones]. AB - In the last 3 years, 300 consecutive patients (110 men, 190 women) were treated for gallstone disease using either traditional open surgery or the video laparoscopic approach. The relative clinical data and results were compared and analysed. The Authors conclude that video laparoscopy presents a number of advantages such as minor costs, reduced pain, quick return to work, compared to traditional surgery, especially when correctly indicated. PMID- 9206500 TI - [Laparoscopic correction of laparoscopic fundoplication failures: 4 cases]. PMID- 9206502 TI - Late arteriovenous fistulas and pseudoaneurysms of the extremity secondary to civilian firearms. AB - To evaluate the treatment of late sequelae of penetrating arterial injuries associated with civilian firearms, such as false aneurysms and arteriovenous fistulas (AV-Fs) of the extremities, a retrospective clinical study was carried out. Whenever diagnosis or operative management is delayed because of local fibrosis or morphological and structural vessel changes, primary repair is not feasible and segmental resection is required. Twenty-four patients presenting late false aneurysms (14) and AV-Fs (11) of the extremities, caused by firearms (6 rifle wounds, and 18 pistol wounds), were identified between 1983 and 1995. Twenty-three patients were submitted to resection of pseudoaneurysms (13) or AV Fs (8), while arterial patch angioplasty was applied in two patients. The venous lesions were repaired by an interposition graft (7) or lateral venorraphy (3). Saphenous vein (15) or PTFE (6) grafts were placed in injured arteries. Percutaneous embolization by coils was performed in two patients. Twenty-three out of 24 patients (95.8%) recovered; one patient (4.1%), affected by aneurysmal AV-Fs of a lower limb dating from 12 years, with enlarged and weakened artery, bled to death. No limb loss occurred. A better evaluation of minimal vascular injuries is required to avoid late sequelae of civilian firearms lesions, with significant blood loss, which need segmental excision of the injured vessels. PMID- 9206501 TI - [Laparoscopic cholecystectomy and respiratory function]. AB - A prospective analysis of 58 patients undergoing laparoscopic cholecystectomy was undertaken to assess pulmonary function changes related to the surgical procedure. Pulmonary function tests were performed preoperatively, as well as 24 hrs., and 48 hrs. after operation. Additionally blood gas analysis was assessed 1 hr. and 6 hrs. after the procedure. At 24 hrs. from operation significant decreases in forced vital capacity, forced expiratory volume in 1", and Tidal volume to 72% (p < 0.001), 70% (p < 0.001), and 92% (p < 0.001) of preoperative values respectively were registered. There was a reduction in PaO2 (p < 0.001) and an increase in PaCO2 (p < 0.05) at 24 hrs. An increase in PaCO2 was also observed 1 hr. (p < 0.001) and 6 hrs. (p < 0.05) postoperatively. Blood gas analysis revealed pH value slightly lower than normal at 1 hr. from operation (p < 0.01). The pulmonary function tests returned to preoperative values at 48 hours. PMID- 9206503 TI - [Upper digestive hemorrhage as a rare manifestation of ectopic pancreas with gastric localization]. AB - The Authors report two cases of ectopic gastric pancreas, one of them causing a massive upper gastrointestinal hemorrhage, representing an infrequent complication of the disease. A literature review on the subject was then performed, and the importance of a differential diagnosis of the ectopy versus gastric malignancies, with relevant prognostical and management implications was assessed. PMID- 9206504 TI - Re: An analysis of papers published in the British and European Journals of Orthodontics. PMID- 9206505 TI - [Diagnosis of nesidioblastosis in the adult]. PMID- 9206506 TI - [Pentoxifylline and oral aphthosis in patients with HIV infection]. PMID- 9206507 TI - [Lumbosacral plexus compression disease of gynecologic etiology]. PMID- 9206508 TI - [Myxoma arising from the lateral wall of left atrium]. PMID- 9206510 TI - [Epithelioid sarcoma of the right arm with brain metastases]. PMID- 9206509 TI - [Significant increase in transaminases in a patient with acute calculous cholecystitis]. PMID- 9206512 TI - [Diagnosis of Gilbert's syndrome]. PMID- 9206511 TI - [Calcification of the ear cartilages in a patient with kidney insufficiency: report of a new case]. PMID- 9206513 TI - [Diagnosis of Gilbert's syndrome: current status of the fasting test. Review of the literature]. AB - Gilbert's syndrome is a benign, often familial condition characterized by recurrent but asymptomatic jaundice. AIM: To describe the involvement of the reduced caloric intake test, used as a diagnostic test in Gilbert's syndrome. METHOD: 49 patients were diagnosed of Gilbert's syndrome for 6 years. 39 patients took 400 kcal/day for three days. The unconjugated bilirubinemia levels were measured at 0, 24, 48 and 72 hours. RESULTS: The 82.05% of test were diagnostics at 24 hours (p < 0.001), while it was necessary 48 hours to 100% of tests were diagnostics (p < 0.05). In any case was necessary to determinate the unconjugated bilirubinemia at 72 hours (p < 0.5). CONCLUSIONS: The best diagnostic efficiency of the reduced caloric intake test is at 48 hours, while the 24 hours determination could be considered diagnostic in a big percentage of the cases. It is not necessary the determination at 72 hours in any case. PMID- 9206514 TI - [Xanthogranulomatous pyelonephritis: experience at the General Hospital of Albacete. Report of 16 cases]. AB - A retrospective study of 16 Xanthogranulomatous Pyelonephritis (XGP) cases seen during a 12-years period (1983-1994) at the Hospital General de Albacete was performed. An analysis is made of the clinical features, laboratory data and imaging diagnostic techniques, comparing our findings with those of several series published in the literature. The outcome of this study is that clinical and biological picture of XGP is characteristic: Middle-aged woman admitted to a Internal Medicine Service because a constitutional syndrome, with important weight loss, general poor health, acceleration of erythrocyte sedimentation rate (ESR), anemia, hepatic disfunction and renal destruction imagings, frequently with renal stone and urinary tract infection history. This characteristics can be useful to suspect a XGP. PMID- 9206515 TI - [Axillary-subclavian thrombosis: review of its etiology and features in recent years]. AB - OBJECTIVES: To describe etiology and clinical characteristics of all patients diagnosed with axillary-subclavian vein thrombosis (ASVT) in our center. METHODS: Retrospective study during last 4 years (January 1991 to December 1994). Only those patients with a typical phlebography were included. RESULTS: 42 patients were diagnosed with ASVT. 10 in 1991, 6 in 1992, 10 in 1993 and 16 in 1994. The average age was 56.3 years. About sex, 20 were in men and 22 in women. The right upper extremity was affected in 16 patients, left side was in 24 and 2 were bilateral. Respect the incriminating agent, a central venous line was the most common, finding in 24 patients (57.14%). A local external compression was detected in 9 patients (21.4%), in 5 the ASVT was associated to a hypercoagulability state and finally only 2 patients were diagnosed with spontaneous o effort ASVT (4.76%). In those associated to a central catheter, 8 patients had a subclavian venous line and 6 had a drum catheter. Lung, colon and breast malignancies were the types of tumours more diagnosed. Treatment basically consisted on intravenous heparin, dicumarinics and subcutaneous heparin. Long term results are not valid because most patients haven't post-treatment phlebography. CONCLUSIONS: The possession of a catheter in the subclavian vein and the existence of a malignancy; especially a lung, colon or breast cancer, are the factors more associated with the development of an axillary-subclavian vein thrombosis in our area. The long of the average life span, the development and use of new oncologic treatment, besides the increasing use of the subclavian veins could do that ASVT would be a more frequent disease. In patients with several associated factors, could be used profilactics treatments. PMID- 9206516 TI - [Analysis of hospital mortality at a regional hospital]. AB - The hospital mortality rate in our centre es 2.34% (264 deaths from a total of 11,336 discharges between 1991 and 1993). The most frequent causes are acute myocardial infarction and cerebrovascular accidents, followed in descending order by pneumonia, chronic bronchitis, congestive heart failure, upper GI haemorrhage, GI tumours, liver cirrhosis, lung tumours and arrhythmias. Our analysis reflects a mortality pattern of a rural population with an age pyramid in which 52% of the patients are older than 45 years. The pattern also reflects the little impact of accidents on our mortality. A 87% of the deaths were older than 65 years with a male to female ratio of 1, 6 and a Swaroop index of 93% and 94% for males and females respectively. PMID- 9206517 TI - [Primary adrenal lymphoma: review and report of a case]. AB - We present a case of non-Hodgkins lymphoma located in both adrenal glands, with diminished adrenal reserve and fatal evolution with serious metabolic complications, with hypoglycemia, severe lactic acidosis, hyperuricemia, acute renal failure, hepatic affectation and hemogram alterations. Much of these complications can be explained by tumoral lysis syndrome probably prompted by the use of high doses of corticosteroids. Primary adrenal lymphoma is exceptional with only 14 cases described in the literature. In spite of its rarity it should be included in the differential diagnosis of uni or bilateral adrenal masses and an early diagnosis is necessary in order to avoid serious and potentially lethal complications. Percutaneous aspiration biopsy can be a valid method of diagnosis because it can identify specific tumoral antigens. The literature concerning this unusual tumour is reviewed. PMID- 9206518 TI - [Viridans streptococci isolated from cerebrospinal fluid. Clinical significance of 9 cases]. AB - Viridans streptococci (VS) are often isolated from cerebrospinal fluid (CSF). However, the significance of such isolates is poorly understood. In the present study we carry out a retrospective analysis of 9 patients in whom VS were isolated from CSF during a 1-year period at La Paz Hospital. Two patients (22.2%) had meningitis diagnosed through clinical, laboratory and bacteriologic findings. Both patients had predisposition diseases (previous difficult spinal tap, ventriculo-peritoneal shunt). The other isolations were considered as contaminants. Three patients (33.3%) with no VS meningitis had other different serious disease (sepsis without bacteriologic confirmation). VS are isolated with relative frequency from CSF, although they cause meningitis in less than one quarter of the cases (those who have a predisposition disease). In the other cases, VS are isolated as contaminants of CSF and other disease should be search as cause of patient symptoms. PMID- 9206519 TI - [Gingival hyperplasia associated with amlodipine]. AB - Drug-induced gingival hyperplasia is a well documented unwanted side effect within the literature. It has been associated with the use of three different types of pharmaceutical agents, including phenytoin, cyclosporine and calcium channel blocking agents. Amlodipine belongs to the dihydropyridine-derived calcium blocking agents that may cause the side effect of drug-induced gingival hyperplasia. In the present study the possible pathogenic mechanisms, clinical and histologic presentation and therapeutic indications of amlodipine-induced gingival hyperplasia are discussed. PMID- 9206520 TI - [Scimitar sign in a partially anomalous pulmonary venous drainage]. AB - The scimitar sign is an uncommon finding, that shows an anomalous pulmonary vein coursing to the right cardiac border, that drains to the inferior vena cava. Usually it is associated to other cardiovascular or pulmonary anomalies, within the hypogenetic lung syndrome. We present a rare case of a few symptomatic man 46 years-old, with partial anomalous pulmonary venous drainage, diagnosed upon finding the scimitar sign in a roentgenogram of the chest, in absence of the hypogenetic lung syndrome. We comment the role of CT in the diagnosis. PMID- 9206521 TI - [The spectrum of bronchiolitis]. AB - Over the last years numerous entities affecting small airways of respiratory tract has been described. It has gone accompanied of a considerable degree of confusion, leading to gather together different diseases which only share the term bronchiolitis. Our aim is to report three patients, one bronchiolitis, noticing their characteristic features and analysing the origin of this situation. PMID- 9206522 TI - [Home care for the terminally ill HIV patient]. AB - Until now, the care of HIV patients, essentially were in the hospital; but the increase in the number of this and the news aspects of them have conditioned changes in the form of assistance. The primary assistance of this patients include prevention, precocious diagnostics, control of assymptomatics patients and diagnosticand-treatment of complications. The last important aspect is home care of terminal patients whose the hospitable assistance is not beneficial as soon as the family support. PMID- 9206523 TI - [Colchicine treatment for recurrent pericarditis]. AB - The treatment of the recurrent pericarditis is difficult habitually. The colchicine, antiinflammatory drug used in gouty arthritis, is effective in preventing the recurrences of acute pericarditis. We review the literature about the efficacy of colchicine in the prevention of recurrent pericarditis. PMID- 9206524 TI - [The conflict between individualized and group care]. PMID- 9206525 TI - [Depression in older persons. Associated factors]. AB - OBJECTIVE: To determine the proportion of elderly people with depressive disorders and study the possible association with sociodemographic factors, self perception of health, cognitive function, diseases suffered, drug consumption, sleep disorders and use of services. DESIGN: An observational crossover study using a home interview. SETTING: Community-based. PARTICIPANTS: 787 elderly people aged 65 and over, not institutionalised and living in the city of Albacete. MEASUREMENTS AND MAIN RESULTS: A questionnaire designed for the study was used to gather data on the sociodemographic variables, self-perception of health, diseases suffered, drug consumption, cognitive function, sleep disorders and contacts with the health service. The variables found by logistic regression to be associated independently to the presence of depressive disorders were: being female (OR = 2.75), habitually suffering sleep disorders (OR = 2.75), having self-perception of poor health (OR = 17.61) and cognitive deterioration (OR = 2.45). CONCLUSIONS: It would be advisable to apply a screening test to detect depressive disorders in elderly people with associated factors (being female, having sleep disorders, self-perception of poor health and cognitive deterioration), so that they could benefit from early diagnosis and adequate treatment. PMID- 9206527 TI - [The physician-patient relationship and the clinical interview (II): the opinion and preferences of physicians]. AB - OBJECTIVE: To find the satisfaction and preferences of Primary Care doctors about the doctor-patient relationship and the clinical interview. DESIGN: Observational, descriptive and crossover study. SETTING: Primary Care. PATIENTS AND OTHER PARTICIPANTS: 51 doctors from 6 health centres in the province of Jaen and 21 family medicine residents. INTERVENTIONS: A validated questionnaire on job satisfaction, the clinical interview and doctors' relationship with their patients, and 25 items to assess the importance of different aspects of doctor patient interaction. MEASUREMENTS AND MAIN RESULTS: 14% (SE, 4.1) of those polled stated they were very satisfied; 70% (SE; 5.4), satisfied; and 11% (SE, 3.7), that the relationship and process of communication with patients was unsatisfactory or very unsatisfactory. Among the most valued aspects were those referring to communication in the interview (information on treatment, use of understandable words, clarification of doubts and summary of the essential points at the end), encouragement to carry out the treatment, correct professional practice (clinical history) and ease in the consultation. CONCLUSIONS: There was great satisfaction with the relationship. The importance of providing understandable information was emphasised. Features of the consultation which focused on the patient were more negatively valued. PMID- 9206526 TI - [The physician-patient relationship and the clinical interview (I): the opinion and preferences of users]. AB - OBJECTIVE: To find user satisfaction and preferences in the doctor-patient relationship and the clinical interview. DESIGN: Observational, descriptive, crossover study. SETTING: Community-based (city of Jaen). PATIENTS AND OTHER PARTICIPANTS: 286 people over 18, randomised with quotas for age and gender. INTERVENTIONS: Validated questionnaire including basic data, whether they are satisfied with their General Practitioner (GP) and 25 items to evaluate the importance of various aspects of the clinical interview and the doctor-patient relationship. MEASUREMENTS AND MAIN RESULTS: 31% (SE, 2.7) of those surveyed were very satisfied with their GP; 37% (SE, 2.8) satisfied; and 4% (SE, 1.1) stated that their relationship was unsatisfactory. Satisfaction, the same as the remembering and carrying out of medical advice, is greater (p < 0.01) in older patients, those with lower social and economic position, those registered at Health Centres and those who saw their GP recently. Those surveyed considered the most valued features of a visit to the doctor to be then items to do with the information provided by the doctor. CONCLUSIONS: A high level of satisfaction with the GP was found, especially from patients registered at a Health Centre. Communication in the interview was highly valued. There was less value placed on features of the consultation which focused on the patient. PMID- 9206528 TI - [An analysis of the diagnoses and prescription for chronic patients at a health center]. AB - OBJECTIVE: To find out if there are differences between inhabitants ages 14-64 with prescription charge (group A), pensioners ages 14-64 without prescription charge (group B), and pensioners aged over 64 without prescription charge (group C), related with chronic morbidity, associated treatment and costs data. DESIGN: A crossover descriptive study (1995). SETTING: Urban primary health care centre. PATIENTS AND OTHER PARTICIPANTS: Participants are inhabitants 14 and over assigned to this health centre (12,605), included in a data bank register, patients are participants with at least a chronic diagnostic (7,007). MEASUREMENTS AND MAIN RESULTS: Participants data were transferred to a practice computer system. Three groups (above) were established according to age band and prescription charge status: group A (73.6% participants), B (8.4%), and C (18%). CONCLUSIONS: Inhabitants ages 14-64 without prescription charge were a differentiated group related with number of chronic diagnoses, number of items to chronic treatment prescribed, and annual average chronically drug costs. So they are intended for use in the future in allocation of budgets to primary health care centres. PMID- 9206529 TI - [The Program of Preventive Activities and Health Promotion. Quo vadis?]. PMID- 9206530 TI - [Noncompliance with the medical prescription in primary care in a rural setting]. AB - OBJECTIVE: To assess the level of initial non-compliance with treatment. DESIGN: An observational study with interviews of patients, which compared prescriptions filled at the Siruela Health Centre (Badajoz) with those dispensed at the town's only pharmacy. SETTING: Primary Care; rural ambit. RESULTS: 8,100 prescriptions were filled for an attending population of 3,100 people. 218 (2.7%) were not collected at the pharmacy. Non-compliance was considerably greater among active workers (4%) than pensioners (2%) (Z = 5.3; p < 0.001). Non-compliance was also greater when prescriptions were written by locums at weekends or on bank holidays (8.6%) than when they were written by the normal doctor (2.2%) (Z = 9.8, p < 0.001). 50.9% of prescriptions not collected were for pensioners on Social Security and 49.1% for the active population. Causes of non-compliance indicated by the patients were: medicine had little effect (33%), high price (28.4%) and not financed by the National Health System (26.6%). 32.6% of the cases of non compliance were followed up; 64.8% of them returned to the consultation. CONCLUSIONS: Better information to patients on their pathologies and treatments would avoid many cases of non-compliance. PMID- 9206531 TI - [The consumption of psychoactive drugs in primary care]. AB - OBJECTIVES: To determine the prevalence of the consumption of psychiatric drugs (PD) among people attending the clinic and to relate that consumption to their sociodemographic characteristics and the presence of psychiatric symptoms. DESIGN: Crossover descriptive study. SETTING: Primary Care. PATIENTS: 350 patients over 14, chosen at random from among those who attended the clinic over an eight-week period. INTERVENTIONS: 1) Questionnaire on sociodemographic data and PD consumption; 2) Self-filling of the GHQ-60 (cut-off point 10/11); and 3) Review of the clinical records to determine the kind of PD, dosage, prescriber, chronic illnesses and the number of consultations over the previous year. RESULTS: 301 (86%) completed the study, 21% consumed PDs. 82% were women. The most consumed PDs were: benzodiazepine (74%) and anti-depressives (34%). CONCLUSIONS: There is a high percentage of PD consumers among people who attend the clinic, especially among women, elderly people, the chronically ill, people living alone and those inactive outside the home. The most commonly used pharmacological group was the benzodiazepines. Almost half the patients had psychiatric symptoms, especially those who lived alone and had no activity outside the home. PMID- 9206532 TI - [Pressure ulcers from the primary care viewpoint: a challenge for everyone]. PMID- 9206533 TI - [Generalized pruritus in primary care]. PMID- 9206534 TI - [Topical antihistamines. Till when?]. PMID- 9206535 TI - [Serological markers in a pregnant population]. PMID- 9206536 TI - [A long-term pharmacoepidemiological study on drug dispensing by the pharmacy office in rural primary care. The influence of age and sex]. AB - OBJECTIVES: To obtain data on the use of medicaments dispensed through pharmacies in primary rural care. DESIGN: Long-term descriptive observational study. SETTING: Nivar and Guevejar municipalities, Sanitary Health District in north Granada (Spain). There were 2.7% losses. PATIENTS: A randomised sample of 589 inhabitants, 18 years o more, included in the census of two villages. MEASUREMENTS AND MAIN RESULTS: The number of visits to the pharmacy, number of medicaments dispensed, prescribing physician and adverse effects to medicaments were recorded and compared between age groups, and between men and women. Prescriptions accounted for 75% of all medicaments, and 67.4% of the prescriptions were written by the primary care physician. Greater numbers of prescriptions were written for elderly persons and women than for younger person and men. Adverse reactions occurred in 3.3% of all subjects. Most reactions were mild (93.6%), and the greatest number of reactions were caused by nonsteroid antiinflammatory agents. CONCLUSIONS: Age and female sex influence the consumption of medicaments. Long-term studies of dispensation patterns in pharmacies can represent a useful method. PMID- 9206537 TI - Abnormal expression of epidermal growth factor and sulfated glycoprotein SGP-2 messenger RNA in a rat model of autosomal dominant polycystic kidney disease. PMID- 9206538 TI - [Cytomegalovirus infections in the course of liver transplantation. Demonstration of viral genome (CMV) using in situ hybridization]. PMID- 9206539 TI - [Pleomorphic liposarcoma of the anterior mediastinum]. PMID- 9206540 TI - [Development of a model for explaining health inequality]. AB - The article proposes a model for systematizing the discussion about the mechanisms of production and reproduction of health inequalities. It starts from existing explanation approaches for combining social and health inequalities in West German literature, from contributions of stress and health lifestyle research and from internationally discussed approaches. The proposed model tries to integrate possibilities of explaining different existing approaches. PMID- 9206541 TI - [Health monitor (SERMO study)--concept, methodology and a paradigmatic result on subjective morbidity in headache]. AB - In Austria, microcensus surveys on self-reported morbidity are carried out at regular intervals every ten years, generally by the Federal Statistic Centre. In the following, we describe an epidemiological observational service which could be regarded as an additional public health instrument. This service is termed "health monitor", and the SERMO (self-reported morbidity) study is the scientific project associated with it. The "health monitor" data provide information on the prevalence of various illnesses and impairment, characteristics and variables of background morbidity by repeated short-term representative surveys on self reported morbidity. The health monitor permits continual observation of the background morbidity of an entire population, while scientific questions pertaining to the SERMO study can be investigated via the health monitor data base. Self-reported morbidity data provide important information about the health of a general population, in addition to clinical and mortality data, and help to make decisions in health policy. By collecting informations (e.g. nearly every month) on background morbidity, "health monitor" and SERMO project could complement other Austrian public health systems to measure the overall health of the population in general. PMID- 9206543 TI - [Quality assurance in child and adolescent public health care]. AB - The most important precondition for quality control in public youth health care is the determination of the required functions. The legal and actual circumstances in the Bundeslander and in the communities and districts vary greatly. Therefore, community youth health care should be supervised by a local committee. Secondly, the description of youth health care products should include complete quality requirements. Thirdly, the structural conditions of production should be analysed. Finally, we should establish audits on outcome quality. PMID- 9206542 TI - [Paradigm shift in dental care services]. AB - According to new legislation socially insured people born after December 31, 1978 do not get prostheses any longer. This law will accelerate the structural change in German dental care that already started in the early eighties. By this health policy finally gives priority to preventive instead of prosthetic dentistry. Some of the most important implications of this paradigm shift for patients and dentists are discussed. PMID- 9206544 TI - [Comparative study of expert assessment of applications for inpatient care according to SGB XI with reference to hospital patients]. AB - The applications for care in a nursing home, proposed by patients in hospitals, should be judged promptly. Judgements based on specifications compiled by the hospital social service are compared to the results of direct examination of the patient in hospital; 96% agreement regarding need for care, 91% agreement regarding necessity of care in a nursing home. We found in 35% a variation in care rank between both methods. In 52% of cases there was no need for a further examination; the care rank was estimated as definite. PMID- 9206545 TI - [Experience in applying legal conditions according to the nursing insurance regulation--ambulatory rehabilitation, nursing aids and nursing care eligibility]. AB - The Statutory Health Care (Nursing) Insurance in Germany, called "Pflegeversicherung (PVG)", financially supports nursing care at home and in nursing homes. If suggested by an authorized examining person (MDK), it provides access to ambulatory rehabilitation and to various aids to facilitate nursing care of patients cared for at home. Also, the PVG mandatory requires the visit by a nurse every three to six months (nursing care checkup) if the patient is cared for by family members without support of professional nursing services. Little is known about the realisation of the suggestions given by the examining person from MDK and about the realisation of the nursing care checkup in actual practice. Our study tried to find answers to these questions. For this purpose, we visited 150 patients at home and asked them about rehabilitation efforts, whether the recommended additional aids had been supplied, and whether nursing care checkups had been successfully performed. RESULTS: For 41 patients, rehabilitatory exercises were suggested, but only 7 were actually realised. 89 patients were supposed to receive various aids, but only 31 were actually delivered. Whenever rehabilitation was performed, it was nearly without exception thanks to the initiative of relatives supported by their private doctor. Nursing care checkups should have been performed in 80 patients, but had been realised in only 11 cases. The care providing relatives considered such "care checkups" to be ineffective, costly and useless. CONCLUSION: The nonfinancial services granted by the PVG were not translated into reality in most instances, and if so, it was due to efforts of relatives and private doctors, not to efforts of the statutory bodies. Apparently this task was more than these bodies could handle during this early phase of services by the PVG. Therefore, the private doctors (family doctors) should be informed of the results of the examination done by MDK. The need for the nursing care checkup should be reconsidered. PMID- 9206547 TI - [Distribution of methadone by pharmacies--the Hamburg method]. AB - This article presents the results of a study in Hamburg about the dispensation of methadone in pharmacies. Experience in Hamburg (since 1988) shows that this "Pharmacy-model" is not only flexible, but also user-friendly. Furthermore the fear that methadone-patients would steal, use violence or harass not only the pharmacist but also other customers proved to be wrong. These experiences call for a reform of the drug-laws permitting the dispensation of methadone in pharmacies in Germany, as it is already practice in some other European countries. PMID- 9206546 TI - [Foreign laborers in Germany--an evaluation of occupational health screening of foreign laborers for determining health disorders]. AB - The following article reports on an analysis of 257,064 medical examination data provided by the German Industrial Engineering and Traffic Safety Control organization, a service of industrial medicine. In addition to the occurrence of diseases (ICD-classification), correlations between the age, sex, nationality and occupations (ILO-classification) are shown, including occupational importance and need for continuing medical treatment. It could be demonstrated that there are different areas of regarding diseases when comparing foreign and German workers, based on places of work, and on social and cultural factors. PMID- 9206548 TI - [Comments on the 2nd GKV revised regulation and immediate program application at the 4 December 1996 hearing]. PMID- 9206549 TI - Bibliography. Current world literature. Cognitive neuroscience. PMID- 9206550 TI - [Research in gynecology today]. PMID- 9206551 TI - [HIV and pregnancy: risks of transmission and therapeutic possibilities]. AB - In Europe, transmission of HIV-1 during pregnancy occurs in 14% of children born to HIV-infected women. Risk factors for transmission are (1) virus load measured by p-24 antigenemia and HIV RNA level, (2) low CD4+ lymphocyte counts (below 600/microliter, (3) placental membrane inflammation and (4) time interval between membrane rupture and delivery. Breast feeding and vaginal delivery increase the risk of transmission of HIV infection. Antiretroviral therapy with zidovudine (Retrovir) at a dose of 500 mg/day reduces the transmission of HIV infection by two thirds. No malformation of the newborn due to zidovudine has been reported so far, but the possibility of unknown long-term adverse effects on children exposed to zidovudine must be weighed against the benefit of a considerable decrease in HIV transmission. Pregnancy is not associated with a higher rate of progression to AIDS, and HIV infection has no adverse effect on the pregnancy outcome in asymptomatic women. PMID- 9206553 TI - [Labor-induced bladder injuries: historical observations]. AB - Injuries to the urinary bladder with development of a fistula during birth were first mentioned around 1030 AD in the opus called 'Al-Kanoun' by the Arabic physician and philosopher Avicenna (Ali Ibn Sina). The observations of D.E. Derry in the mummy of Henhenit seem to have made sure that this obstetric complication already existed earlier on. Henhenit lived at the court of king Mentuhotep II (around 2050 BC). During the second half of the 19th century injury-related and necrosis-related fistulas were distinguished for the first time. Jobert de Lamballe (1852), Marion Sims (1852), and Gustav Simon (1854) created the basis for successful operative treatment of vesicovaginal fistulas. PMID- 9206552 TI - [Value of premedication with gonadotropin releasing hormone agonists before transcervical resection of solitary submucous myoma]. AB - In a retrospective study, the value of pretreatment with a gonadotropin-releasing hormone (GnRH) agonist before hysteroscopic myoma resection was assessed in 31 patients suffering from menorrhagia. No difference in operating time, intra operative fluid deficit, blood loss and postoperative eumenorrhoea rate was found between pretreated and non-pretreated patients with solitary submucous myomas smaller than 4 cm in diameter. Thus, pretreatment with GnRH agonists before the resection of small solitary submucous myomas has no benefit for the patients. PMID- 9206554 TI - [History of menstruation--an aspect of the medical history of the woman]. AB - The understanding of menstruation as well as the image of women have much changed in the course of history. This development, as reflected by the views of the Old Testament (Leviticus), of Hippocrates and Aristoteles, its characterization in the books of Hildegard of Bingen and of Paracelsus, its description in the Renaissance and the 18th century, is followed up to our modern times. PMID- 9206555 TI - [Triplet pregnancy after intracytoplasmatic sperm injection of cryopreserved testicular sperm]. AB - For the first time we report on an intact and ongoing triplet pregnancy after intracytoplasmatic sperm injection of cryopreserved testicular sperm. Indication was azoospermia due to hypergonadotropic hypogonadism. The patient conceived in the third treatment cycle after 25 treatment cycles with donor sperm that had been carried out without success in two other treatment centers. PMID- 9206556 TI - [Champagne administered by spoon for peritonitis after cesarean section. From the history of obstetrics in Erlangen--data from about 60,000 deliveries in 100 years]. AB - The University of Erlangen has been engaged in clinical obstetrics for approximately 170 years. During this time, Erlangen University's delivery house, opened in 1828 and at first having considerably less than 50 births a year, developed into a perinatal centre with approximately 1,700 births a year. For the period from 1880 to 1981, a group of MD students reviewed the existing records and evaluated 60,000 births with respect to more than 40 parameters. Part of the results obtained are shown with special reference to operative obstetrics. Apart from the general influence of the scientific development on decisions and results within obstetrics, individual factors were also recognizable, factors which are linked with the experiences, insights and specialized working areas of the particular head of the hospital. PMID- 9206557 TI - [Delusions of witches and suppression of femininity in the Occidental culture from the analytic viewpoint]. AB - In this study, we look into the historical roots of the witch-hunt. For a period of 540 years, between 1241 and 1782, the burning of witches has been well documented. During these 540 years witch-hunts were simply the culmination of a passionate hostility against femininity as such within western culture. As can clearly be seen in the texts of the great medieval theologians, the humiliation and degradation of the female sex--and the condemnation (or diabolization) of sexuality as well as antisexual prejudices--originate from a pre-Christian, heathen-mythological way of thinking, which was largely established by the medieval church as a form of syncretism and the notions of which persist even today. In those days the compulsive regulation of sexuality and the threat of the torments of hell, if one did not obey these rules, initiated the vicious circle of failure, guilt and fear in people's minds and so made it impossible to overcome the Oedipus complex. This regression to the 'anal stage of development' prepared the ground for sadomasochistic destructiveness, which was (supported by superstition) projected outwards onto the so-called witches. PMID- 9206558 TI - [Ritanserin prolongs the analgesic effect of morphine and slows the development of tolerance]. AB - The effect of 5-HT2 receptor blocker ritanserine on the analgesic effect of morphine was studied in experiments on mice. A single simultaneous injection of ritanserine (10 mg/kg) and morphine (1, 10, and 20 mg/kg) prolonged the duration of analgesia in the tail clip test. Combined subchronic (6 days) injection of morphine and ritanserine (1, 5, and 10 mg/kg) twice a day delayed the development of tolerance to the opiate analgesic effect in the hot plate and tail clip tests. PMID- 9206559 TI - [The effect of CO2 on conduction blocking in nerve fibers by trimecaine and anestezin]. AB - It was shown in experiments on the sciatic nerve of the Rana ridibunda frog that CO2 promotes blocking of stimulation conduction in the nerve fibers by the tertiary amine trimecaine but does not change the rate of blocking by the neutral anesthetic anesthesin. The increased rate of trimecaine blockade development is explained by decrease of cytoplasmic pH under the effect of CO2, which leads to increase in the number of charged molecules of the anesthetic entering the sodium canal and blocking it. PMID- 9206561 TI - [The cardiotoxic properties of anti-arrhythmia agents]. AB - Experiments were conducted on cats to study the cardiotoxic and arrhythmogenic properties of some antiarrhythmic agents in continuous intravenous infusion in doses multiple of the dose which causes an antiarrhythmic effect on an acontic model of arrhythmia in rats in 50% of cases. Novocainamide, cordarone, bonnecor, and quinidine possessed the most marked cardiotoxicity. A less marked cardiodepressive activity was encountered in the case of ethmosine and ethacysin. The appearance of the cardiodepressive properties of the agents depended on the rate of their injection. The results of the study testify to the possibility of prognosticating the arrhythmogenic and cardiodepressive effects in experimental investigation of new antiarrhythmic agents. PMID- 9206560 TI - [A preclinical evaluation of the pharmacological activity of a mixed potassium and magnesium glutamate]. AB - Mixed magnesium and potassium glutamates with different ion ratio are synthesized. The antiarrhythmic activity of the compounds obtained is studied on the models of strophantin, calcium chloride, aconitine arrhythmia, and arrhythmia after Harriss. It is found that magnesium and potassium glutamate of the composition KMg(HGlu)4.4H2O exhibits higher activity compared to glutamate of the composition KMg(HGlu)4.3H2O on the strophantin and calcium chloride arrhythmia models. The study performed on the strophantin and calcium chloride models as well as on the Harriss arrhythmia model showed that KMg(HGlu)4.4H2O is more effective as antiarrhythmic drug than asparkam and panangine but, at the same time, the effect of KMg(HGlu)4.4H2O within the aconitine model is somewhat less pronounced compared to panangine. PMID- 9206562 TI - [The anti-arrhythmic effect of the selective sigma-receptor antagonist DuP 734 [1 (cyclopropylmethyl)-4-(2'-(4''-fluorophenyl)-2'-oxoethyl)piper idine HBr]]. AB - It was demonstrated on models of epinephrine-, CaCl2-, and aconitine-induced arrhythmias that the sigma-receptor antagonist DuP734 possesses high antiarrhythmic activity in a dose of 1 mg/kg. The sigma-receptor agonist (+)SKF10,047 promoted the development of ventricular fibrillation. Atropine had no effect on the antiarrhythmic properties of DuP734. "Chemical sympathectomy" of the myocardium with the ganglion-blocking agent hexamethonium also did not affect the proarrhythmic properties of (+)SKF10,047. It is supposed that selective sigma receptor blocking agents may be highly effective antiarrhythmic drugs. PMID- 9206563 TI - [The effect of sodium nitroprusside and nitro-containing vasodilators on rat liver microsomal mono-oxygenases]. AB - It has been shown in vitro that sodium nitroprusside (SN) causes a concentration dependent suppression of the enzymatic activity of aminopyridine demethylase APDM(IC50 = 1.86 x 10(-4) M) and ethoxycoumarin O-deethylase-ECOD(IC50 = 1.46 x 10(-4) M) in hepatic microsomes of Wistar rats, which is related to different isoforms of cytochrome P-450. The inhibiting effect of other nitro compounds (nitroglycerin and isosorbide dinitrate) in relation to APDM is less marked than that of SN. These compounds have practically no effect on ECOD activity. It is assumed that the degree of the inhibiting effect of SN, nitroglycerin, and isosorbide dinitrate is connected with the mechanism of NO release. PMID- 9206565 TI - [A method for stimulating the protective mechanisms of the gastric mucosa in peptic ulcer]. AB - It was shown on an experimental model of chronic gastric ulcer in rats that administration of microsomal oxidation inductors benzonal (benzobarbital) and phenobarbital in a dose of 50 mg/kg for 10 days significantly increases the content of cytochrome P-450 in the gastric mucosa. The increased content of P-450 promotes stimulation of the synthesis of mucosal barrier glycoproteins. The cytoprotective effect of famotidine in a dose of 100 mg/kg and sucralfate in a dose of 400 mg/kg proved to be much weaker than that of the inductors. PMID- 9206564 TI - [Buspirone--a wide-spectrum preparation]. AB - It has been demonstrated on albino rats that buspirone produces an antidepressive and nootropic-like effect in small doses (0.5-1.0 mg/kg) and an anxiolytic, cataleptogenic, and neuroleptic-like effect in high doses (5.0-10.0 mg/kg). The wide spectrum of its activity is due to the presence of various mediator components, serotonin- and dopaminergic in particular, in the mechanism of its action. PMID- 9206566 TI - [Fenibut and fenazepam as potential protectors of pregnancy with a GABA-ergic mechanism of action]. AB - It has been shown that the GABAergic system of the myometrium takes part in regulation of uterine contractility during the development of pregnancy Fenibut and phenazepam exhibit a specific uterotropic effect. The uterodepressant effect of fenibut is realized via the inhibiting GABA-receptors of the myometrium. The uteroinhibiting effect of phenazepam is mediated through the interaction with the postsynaptic GABA-benzodiazepin-C1-receptor complex of the myometrium. PMID- 9206567 TI - [The effect of the chronic internal administration of the new Russian n-3 polyunsaturated fatty acid concentrate--epaden--on blood coagulation system indices in rabbits]. AB - Daily oral 6-week administration of epaden in a dose containing 0.3 g eucosopentanoic acid and 0.05 g docosahexaenoic acid caused decrease in collagen induced platelet aggregation in rabbits in vivo and in the activity of the tissue type plasminogen activator, as well as reduction in the level of antithrombin III cofactor activity. No changes were encountered in ADP-induced aggregation, in the platelet count, in platelet adhesion to collagen, and in activated partial thromboplastin time. PMID- 9206569 TI - [Lidocaine as an immunomodulator in a toxic liver lesion]. AB - D-galactosamine (DGA) increases the malonic dialdehyde (MDA) content in the erythrocytes, reduces the ATP content, and induces the appearance of immunosuppressive properties in the red cells. Administration of lidocaine attenuates or completely removes these effects of DGA in poisoned animals. Extracorporeal treatment of the erythrocytes of intact rats with blood serum of DGA-poisoned reduces the ATP content and induces the appearance of immunosuppressive properties in the erythrocytes. Blood serum of DGA-poisoned rats which had been given lidocaine does not cause such effects. PMID- 9206568 TI - [The effect of heparin on the activity of the mono-oxygenase glucuronyl- and glutathione transferase systems in ultrasonic damage to the rat liver]. AB - Six days after local exposure of the rat liver (during operation) to ultrasound (2 W/cm2, 1 min), the cytochrome P-450 content in the microsomal fraction reduced, and the rate of NADPH oxidation, the NADPH cytochrome P-450 reductase activity, ethylmorphine demethylation and aniline hydroxylation decreased in 12 days the activity of NADPH-cytochrome b5 reductase and cytosol sulfobromophthalein-glutathione-S-transferase also reduced. Heparin administration (250 U/kg i.m. every other day 3 and 6 times) had an enzyme activating effect (particularly in the late-term restoration period) in animals which had been exposed to ultrasound. PMID- 9206570 TI - [The effect of stimulants of the mononuclear phagocytosing system on protein synthesis in rat organs and tissues]. AB - It has been demonstrated in rat experiments that stimulation of the mononuclear phagocytizing system with a polysaccharide intensifies protein biosynthesis in various organs and tissues. In administration of high-polymer RNA into the organism increase of protein synthesis was encountered only in immunocompetent and blood-forming tissues. It is concluded that a stimulating effect in relation to macrophages is characteristic of high-polymer compounds which are absorbed by them due to receptor mediated endocytosis. The differences in the receptor apparatus of the resident macrophages determine the discovered difference in the intensification of protein biosynthesis in the organs and tissues of rats. PMID- 9206572 TI - [Adenosine pharmacokinetics in rats]. AB - The pharmacokinetics of the effective peripheral vasodilator adenosine in blood serum of rats was studied after its intravenous injection. The dynamics of labeled adenosine in blood was described by means of a one-compartment mathematical model. The pharmacokinetic parameters of the drug were calculated from the model coefficients. The distribution of the label of adenosine and its metabolites in blood serum and heart, liver, and kidney tissues was studied by thin-layer chromatography. PMID- 9206571 TI - [The pharmacokinetics of the dipeptide analog of piracetam with nootropic activity GVS-111 and of its basic metabolites]. AB - The pharmacokinetics of a new nootropic dipeptide analog of piracetam-N phenylacetyl-L-prolylglycine (GWS-111) and its main metabolites were studied in rats by means of high performance liquid chromatography and gas-liquid chromatography. The compound under study showed a greater resistance to an enzymatic effect than natural neuropeptides. In addition to an unchanged compound three of its metabolites were found in the blood plasma of the rats. One of them, cyclo-Pro-Gly was an active metabolite of GWS-111. PMID- 9206573 TI - [A comparative study of the permeability across the hemato-encephalic barrier and of the effect on cerebral blood flow of the new neurotropic agent calcium ketohomopantothenate and of pantogam]. AB - The permeability through the blood-brain barrier of N-(4-hydroxy-3,3 dimethyl-2 oxo-1-butiryl) gamma-aminobutyric acid calcium salt, a new neurotropic drug calcium ketohomopantothenate (KPA-Ca), was studied in comparison with that of calcium homopantothenate, pantogam (P) Liquid chromatography analysis showed that after oral administration of KPA-Ca and P both forms of these agents, namely, oxy and keto- derivatives of homopantothenic acid, were found in the brain of experimental rats, the KPA-Ca (ketoform) content being higher. Pharmacological studies showed that KPA-Ca penetrates the blood-brain barrier in greater amounts and causes a higher effect on the rate of cerebral blood flow when it is administered per os in a dose of 50 mg/kg. PMID- 9206574 TI - [The effect of epsilon-aminocaproic acid and prednisolone on the activation of the complement system due to x-ray contrast agents in rats in vitro]. AB - epsilon-Aminocaproic acid and prednisolone in concentration of 3.0 x 10(-5) M are proven to reduce or even completely prevent activation of the alternative pathway of the component which has been induced by iod-amide adipiodone and triombrast in concentrations 2.5 x 10(-2) M in the serum of "sensitive" rats. The preventive effect of the glucocorticoid appeared less pronounced. There exists direct positive correlation between the degree of the component activation with radiopaque compounds and preventive effect of the drugs studied. PMID- 9206575 TI - [The toxicological characteristics of ammonium glycyrrhizinate (glycyram). A study of its acute and subacute toxicity]. AB - A single parenteral and oral administration of ammonium glycyrrhizinate in rat and mice experiments showed that the compound is related to practically nontoxic drugs. Its repeated administration (30 times) into the stomach in a maximum daily therapeutic dose (7 mg/kg) and in a four-fold dose (28 mg/kg) did not cause signs of intoxication, essential changes in the hematological and integral parameters, shifts in the activity of serum enzymes, morphological changes in the cell structures of the internal organs. Administration of the drug in a dose of 28 mg/kg for a second time led to changes in the activity of some enzymes in the brain, the development of parenchymatous dystrophy of the liver which changed to acidophilic necrosis attended with signs of regeneration. Under conditions of a subacute experiment the maximum daily therapeutic dose of ammonium glycyrrhizinate may be considered practically nontoxic. PMID- 9206576 TI - [The effect of nicotinamide, methionine and alpha-tocopherol on the liver conjugating and mono-oxygenase systems and on lipid peroxidation in hepatosis hepatitis in rats]. AB - Four days after a single intragastric injection of CCl4 (5 mg/kg as a 50% oil solution) increased intensity of a chemoluminescence "quick flush" in the hepatic microsomes and blood serum, as well as hepatocyte cytolysis (increased ALT activity) and decreased rate of antipyrine elimination from the blood were recorded in rats. The content of cytochromes P-450 and b5 activity of NADH cytochrome b5 reductase and cytosol glutathione transferase in the hepatic microsomal fractions reduced in this case. Administration of methionine (200 mg/kg) and its combination with nicotinamide (60 mg/kg) without and, particularly, with additional prescription of vitamin E (150 mg/kg) produced a marked antioxidant and enzyme-normalizing effects in the rats. PMID- 9206577 TI - [Xanthurenic acid inhibits the activity of an experimental epileptogenic focus in the rat hippocampus]. PMID- 9206578 TI - [The anti-edematous effect of the preparation polyosm in acute hypertensive encephalopathy]. AB - Experiments were conducted on Wistar rats to study the effect of intravenous infusion of polyosm (solution of polyethyleneoxide 400) on the parameters of brain tissue edema-swelling (according to impedancemetry findings) and the local cerebral blood flow on a model of acute hypertensive encephalopathy. Under such conditions the drug hastened involution of edema-swelling of the brain tissue and prevented a decrease in cerebral blood flow. PMID- 9206579 TI - [A clinico-pharmacological study of olifen in periodontal inflammation]. AB - Clinical study of the efficacy of oliphen treatment in generalized periodontitis in comparison with galascorbin treatment was conducted. Oliphen arrested the inflammatory process in the periodontium quickly and effectively and shortened the duration of treatment by half. Under its effect lipid peroxidation became less intense, the level of malonic dealdehyde decreased, and the activity of catalase in the oral fluid of patients with periodontitis increased. The favorable effect of oliphen in periodontitis is connected with its antioxidant and antihypoxant properties. PMID- 9206580 TI - [The possible means for the pharmacological correction of lipid metabolic disorders in atherosclerosis]. AB - The main trends of contemporary pharmacology of hypolipidemic and hypocholesterolemic agents were analysed. A list of drugs capable of correcting lipid metabolism disorders is given: cholesterol absorption inhibitors, stimulators of bile acid synthesis, inhibitors of cholesterol synthesis, analogs of fibroic acid and other inhibitors and correctors of hypertriglyceridemias, stimulators of reverse cholesterol transport and synthesis of high-density lipoproteins, etc. The latest theoretical and experimental elaborations of gene therapy of dyslipoproteinemias in atherosclerosis are discussed. PMID- 9206581 TI - An open thesaurus for diagnostic codification in practical hematology. PMID- 9206582 TI - Changes in psychologists' salaries. PMID- 9206583 TI - [Processing acoustically presented time intervals of seconds duration: an expression of the phonological loop of the working memory?]. AB - Working memory has been proposed to contribute to the processing of time, rhythm and music; the question which component of working memory is involved is under discussion. The present study tests the hypothesis that the phonological loop component (Baddeley, 1986) is involved in the processing of auditorily presented time intervals of a few seconds' duration. Typical effects well known with short term retention of verbal material could be replicated with short-term retention of temporal intervals: The immediate reproduction of time intervals was impaired under conditions of background music and articulatory suppression. Neither the accuracy nor the speed of responses in a (non-phonological) mental rotation task were diminished under these conditions. Processing of auditorily presented time intervals seems to be constrained by the capacity of the phonological loop: The immediate serial recall of sequences of time intervals was shown to be related to the immediate serial recall of words (memory span). The results confirm the notion that working memory resources, and especially the phonological loop component, underlie the processing of auditorily presented temporal information with a duration of a few seconds. PMID- 9206584 TI - [Multidimensional scaling of color patterns of the DIN color chart]. AB - The DIN color chart was developed in the 1950s by Manfred Richter using of classical psychophysical scaling techniques. It is based upon the idea that colors are ordered along three subjective dimensions, i.e. hue, saturation, and brightness. Furthermore, it is assumed that the colors of the DIN color chart fulfill the principle of specific equidistancy. The main aim of this study was to investigate this claim empirically. More specifically, it was tested whether colors of the same hue and brightness are equally spaced along the dimension of subjective saturation. The data were collected in three paired comparisons experiments and were analyzed using multidimensional scaling. According to the results, the psychological properties of the DIN color system can be replicated approximately. In addition to a confirmation of the postulate of equidistancy this research contributes to the understanding of the convergence of different scaling methods. PMID- 9206585 TI - [Are there differences in processing times of variously complex rules in differential eyelid conditioning?]. AB - Two logical relations, conjunction (AND) and exclusive disjunction (XOR) differ in formal complexity as well as in observable difficulties. AND results in less errors, fewer trials to criterion, and shorter processing time per trial than XOR. Two paradigms of differential classical conditioning are based on these rules. Negative patterning (A+, B+, AB-) equals XOR, and positive patterning (A-, B-, AB+) equals AND. We studied experimentally whether or not differences in processing time per trial are reflected in different optimal interstimulus intervals in human eyelid conditioning. Results of four groups (AND/XOR x 400/1200 ms; each group n = 10) suggest that differential conditioning could be observed in positive patterning (800-1000 ms) earlier than in negative patterning (1000-1200 ms). PMID- 9206586 TI - [Categorization in infancy: differentiation of global object classes]. AB - Two studies tested whether preverbal children distinguish global categories (animal and furniture) on a conceptual basis. A total of 59 eleven-month-olds solved an object examination task. During habituation, infants freely explored different natural-looking toy models from the same category. In Study 1, the same series of four different examplars was presented twice. In Study 2, ten different exemplares were presented. In both cases, a significant habituation effect could be observed. When a perceptually new object of the same category was presented on the first test trial after habituation, a significant increase in examination time from the last habituation trial to the first test trial could be observed in Study 1. When a new object of the contrasting category was presented on the second test trial, examination times increased significantly from the first to the second test trial in both studies. These results support earlier findings suggesting that preverbal infants are able to distinguish global categories on a conceptual basis. PMID- 9206587 TI - [Analysis of interference effects in simultaneous processing of 2 problems]. AB - The aim of this investigation was to analyze dual-task interference in the so called Psychological Refractory Period (PRP) paradigm. In this paradigm subjects have to carry out two choice reaction tasks that overlap in time. A well known result is that reaction time on task 2 (Rt2) increases with decreasing overlap of the two tasks. Thereby reaction time on task 1 is described to be independent of the size of overlap (Rt 1). Usually, this result is explained by the assumption of a PRP which arises in processing of task 2, when serial processing is ongoing in both tasks. It was asked, 1.) whether the PRP is located before or after response selection in the first task and 2.) how the second task influences first task processing. In the experiment subjects had to carry out two choice reaction tasks together. In different conditions the difficulty of response selection in task 2 was systematically increased by varying the number of response alternatives (0, 1, 2, 3). Difficulty of response selection in task 1 was held constant. Overlap between both tasks was varied. This experimental design allows different hypotheses about the sources of interference in both tasks to be examined by use of Schweickert's Critical Path Technique (Schweickert, 1983). Contrary to the results of Karlin and Kestenbaum (1968) the effects of number of response alternatives and size of overlap on Rt2 indicate a localization of the PRP before response selection. The results support models which assume a serial processing in response selection (Welford, 1952). They are contrary to models of parallel processing in this stage (Keele, 1973). The influence of the number of alternatives in task 2 on Rt 1 can be explained by a mechanism of grouping both motor responses. PMID- 9206588 TI - Substrate specificity of carp Cyprinus carpio cathepsin H with methylcoumarylamide substrates. AB - Cathepsin H was purified from the crude extract of carp (Cyprinus carpio) hepatopancreas by a reformed method involving six stages, and the specific activity increased about 11,500-fold with a 23% recovery. Of varying fluorescent synthetic substrates tested, carp cathepsin H possessed an ability to hydrolyze four N-terminal unblocked substrates those are composed of a single amino acid bound to 4-methylcoumaryl-7-amide (MCA), namely Leu-MCA, Arg-MCA, Lys-MCA and Ala MCA. In contrast the enzyme was only marginally able to hydrolyze an unblocked substrate such as Pro-Phe-Arg-MCA and totally unable to degrade all blocked derivative employed. From the kinetic constants with four unblocked substrates, car cathepsin H had the highest affinity toward Leu-MCA with a Km value of 35.4 microM. Besides both the hydrolytic rates and molecular activities of the enzyme decreased from Lys-MCA > Arg-MCA > Ala-MCA > Leu MCA as judged by their Vmax and Kcat values, respectively. Otherwise, optimal pHs for hydrolysis of cathepsin H were different for four substrates. The enzyme exhibited maximum level of the activity at pH 6.5 for Arg-MCA and Lys-MCA and at pH 7.0 for Leu-MCA and Ala-MCA. PMID- 9206589 TI - Organophosphate effects on antioxidant system of carp (Cyprinus carpio) and catfish (Ictalurus nebulosus). AB - The effect of the organophosphate insecticide Dichlorvos on antioxidant enzymes and other oxidative and redox parameters of carp (Cyprinus carpio L.) and catfish (Ictalurus nebulosus) were studied. Changes in superoxide dismutase, catalase, glutathione peroxidase, and in the case of carp acetylcholinesterase activities were studied in tissue homogenates. Other parameters studied: changes of lipid peroxidation, reduced glutathione and the amounts of two radicals, superoxide and hydroxyl radicals are compared. Our results showed that the organophosphate tested, besides its inhibitory effect on acetylcholinesterase--or together with it--induces changes characteristic of "oxidative stress." PMID- 9206590 TI - [Pathology and pathogenesis of secondary epilepsy to hypoxic-ischemic encephalopathies]. AB - The neuropathology of haemorrhagic and hypoxic-ischaemic perinatal encephalopathies and their effect on the post-natal development of the brain, has been studied in children who survived with these lesions (for days, weeks, months and even years). Eventually some children developed neurological sequelae, including epilepsy and cerebral palsy. In this paper it is emphasized that the post-natal development of the grey matter next to these lesions in altered in a specific manner. The post-natal resolution (scarring) of the subpial haemorrhage causes structural changes in the superficial layers of the cortex and permanent leptomeningial heterotopia. The pyramidal cell of layers II and III whose apical dendrites had been partially amputated by haemorrhage became star cells. The grey matter often survived an infarct of the subjacent white matter, since its circulation remained intact. However its post-natal development was altered in a specific way. The post-natal development of this grey matter (partly deprived of sensory information because of the destruction of afferent fibres and with contact lost because on the destruction of efferent fibres) is inevitably different. Projection pyramidal cells (long axon) axotomized by the subjacent lesion, survive the insult and post-natally are changed into intracortical short axon cells. The intrinsic neuropile of the grey matter (partially isolated) increases in an irregular manner which can be seen using immunohistochemical techniques and Golgi's method: areas with a great increase in fibres alternate with areas with few fibres. The presence of large neurones (Golgi's method) with long drendites covered with spines (acquired neuronal hypertrophy) is frequent. In this paper it is suggested that these changes in the grey matter secondary to subpial haemorrhage and hypoxic-ischaemic perinatal infarcts are accompanied by functional changes which may play and important role in the pathogenesis of epilepsy (infantile spasm) and in infantile cerebral palsy. PMID- 9206591 TI - [Benign infantile familial convulsions]. AB - INTRODUCTION: Benign Infantile Familial Convulsions (HBIFC), are characterized by brief partials seizures, occasionally with secondary generalization, with onset in the first year of life, family history of similar electroclinical seizures and same age of appearance. MATERIAL AND METHODS: We presented 16 patients, 10 girls and 6 boys, evaluated in our Service between 1990-1996. We analyzed, age of onset of the seizures, sex, family history of epilepsy, neurologic exam, semiology, distribution, frequency and duration of the seizures, EEG, neuroradiologic studies and evolution. RESULTS: The patients had partial seizures, which occurred mainly in clusters, with onset ranged from 3 to 8 months, with normal neurological exam and psychomotor development. The interictal EEG was normal and the course was benign. Treatment response to antiepileptic drugs was good. CONCLUSIONS: Our presentation confirmed that BIFC are a new partial idiopathic epileptic syndrome, with a genetic predisposition, probably with an autosomal dominant inheritance, which would be recognize in the next international classification of epilepsy and epileptic syndromes. PMID- 9206592 TI - [The evaluation of a newborn in a coma]. AB - Coma is differentiated from sleep by the absence of a normal arousal response and from death by the presence of heart beats and the absence of brain death criteria. Most causes of coma are readily diagnosed and treated. Others require a test whose results are not immediately available, transportation or a risky procedure and empirical treatment has to be considered. In addition to treating the cause of coma, treatment of the systemic and neurological causes of secondary brain damage is paramount. PMID- 9206593 TI - [Intrauterine exposure to drugs]. AB - The development of the fetal central nervous system can be effected by drugs. In this paper we review the neurological consequences of intrauterine exposure to alcohol, cocaine, opiates and marijuana. Ethanol causes the fetal alcohol syndrome: mental retardation, intrauterine and postnatal growth retardation, and peculiar dysmorphic features. Is pathogenesis has been explained on the basis of maternal nutritional deficiencies or due to abnormalities in the conversion of ethanol to aldehyde, or abnormalities in the metabolism of prostaglandins or retinoic acid, the neurotransmitter systems, the neuronal excitotoxic activity, the development of the white matter, the production of gangliosides, and/or genetic regulation cell-cell adhesion. Cocaine has been related to congenital malformations, neurologic abnormalities during the neonatal period and psychomotor and cognitive development deficits. Characteristic dysmorphic features and a higher incidence of the sudden infant death syndrome (SIDS) have also been described. The following mechanisms have been implicated in the pathogenesis: vascular effects, superoxide formation, chelation of calcium ion channels, and abnormalities in the production of glycosphingolipids, the synthesis of DNA, the functioning of neurotransmitter systems, the neuronal growth and differentiation, the neuronal excitotoxic activity and/or the expression of early immediate genes. Opiates produce intrauterine and postnatal growth retardation, neonatal abstinence syndrome, and deficits of the psychomotor and cognitive development. They also increase the incidence of SIDS. The pathogenesis has been related to abnormalities in the sensitivity of the locus ceruleus, the functioning of the neurotransmitter systems, and/or the expression of early immediate genes. Marijuana has been associated with intrauterine growth retardation, dysmorphic features, and abnormalities of the behavior during the neonatal period, the psychomotor and cognitive development, and the sleep. The pathogenesis is thought to be due to an action upon specific receptors, or upon the neurotransmitter systems, and/or to an increase in the production of carbon monoxide. The best treatment of the syndrome of intrauterine exposure to drugs in the prophylaxis. The identification of emotional and drug addiction problems in the mother can avoid disastrous consequences. The care of these children is complex and requires a good pediatric follow-up and an early intervention program while the mother on the parents continue with the drug addiction therapy. The coordinations of all the necessary services with the active participation of social workers, physicians, educators and teachers is crucial for a successful treatment. PMID- 9206594 TI - [Genetic diagnosis of the hypotonic newborn]. AB - Assessment of hypotonic newborn babies implies not only neurological studies, but; also new methods of molecular genetics, to reach a diagnosis of the aetiology. The Prader-Willi, Werdnig-Hoffmann and Myotonic Dystrophy syndromes are three conditions with neurological symptoms which have recently been defined at a molecular level. PMID- 9206595 TI - [Neonatal seizures: the beginning and interruption of the treatment]. AB - Opinions vary as to which 'neonatal seizures' should be treated and which should not be. There is also controversy as to when therapy of seizures should be discontinued. A poll taken among a group of eleven experts in these matters concurred in the opinion that classical neonatal seizures, those well established by clinical and EEG criteria, can be treated before confirmatory EEG support is obtained. They also agreed that those movements known as subtle seizures should await EEG or video/EEG confirmation before commencing therapy. Nine of these experts now believe that therapy for seizures should be terminated at an early time. Most will stop the therapy while the patient is still in the nursery with the exception of 3 who would treat the babies for 3 months. PMID- 9206596 TI - [Neurocognitive evaluation]. PMID- 9206597 TI - [Neurocognitive treatment for learning disabilities]. AB - The present study examines the viability of an intervention model intended to develop cognitive and metacognitive strategies, by sel-instructional procedures. Furthermore the results of programs that explore the efficiency of this model in the interventions for students with learning disabilities (LD) are discussed. Finally, a number of guidelines for the future research in this area are included, underlining the importance of contextualizing the programs aimed in teaching to thinking. PMID- 9206598 TI - [Epilepsy and learning disabilities]. PMID- 9206599 TI - [The early diagnosis of cerebral paralysis]. PMID- 9206600 TI - [Cerebral palsy therapy]. AB - Unfortunately, in spite of the advances in foetal and perinatal medicine in the last twenty years, the incidence of cerebral palsy has remained unchanged (1.5 2.5 per 1000 live births). It has even possibly risen slightly in premature babies of low birth weight, in parallel with the increased survival of these babies. In spite of modern techniques of rehabilitation, 25% of these patients cannot walk and 35% are mentally retarded. This costs society 5,000 million dollars annually, not counting the loss of opportunity and the emotional and economic burden imposed on these families. The development of new preventive measures such as the use of antagonists of the cortical excitatory amino acids (which when in excess may cause irreversible cerebral damage in cases of hypoxic ischaemic encephalopathy of the new born). Intramuscular botulinum toxin and continuous intrathecal baclofen seem to promise a reduction in the incidence and functional incapacity of cerebral palsy. PMID- 9206601 TI - [The neurology of dyslexia]. AB - Developmental dyslexia is a heterogeneous disorder in which the prominent manifestation is a discrepancy between reading achievement and intelligence. There is no serious auditory, visual, psychiatric, social or educational factor that could be responsible of the discrepancy. Males are more often affected than females. It is a pervasive condition but with adequate help and spontaneous compensation, reading ability may improve. Neuroimaging, mainly MRI, allows to demonstrate in two thirds, an absence of the usual symmetry of the planum temporale favoring the left side. Twenty to 25% of the remaining cases show asymmetry of the right side. Etiology is unknown but heredity plays an important role. The pathology of dyslexia has revealed abnormalities of the cerebral cortex focal four-layer microgyria, microdysgenesis and arteriovenous malformations. Galaburda hypothesises that a pre or perinatal adverse event produces a basic cognitive, progressive alteration, that eventually invades the perceptual elements (visual, phonological, semantic-syntaxic difficulties). Heilman, Voeller and Alexander propose a motor-articulatory feedback hypothesis. PMID- 9206602 TI - [Language disorders: is EEG clinically useful?]. PMID- 9206603 TI - [Consequences of recurrent seizures during development]. AB - While many children with recurrent seizures have a good prognosis, a small percentage of children with intractable epilepsy have a more ominous course with a gradual decline in cognitive abilities over time. While the reasons for this cognitive decline may be multifactorial, there is evidence both from human and animal studies that recurrent seizures may lead to gradual cognitive impairment in some children. Laboratory studies have also demonstrated that recurrent seizures can lead to deficits in learning and memory as well as structural changes in the brain. It is important for the clinical to be aware of gradual declines in intelligence that may occur over time. PMID- 9206604 TI - [Risk factors in recurrent seizures and in mental retardation]. PMID- 9206605 TI - [Neuropathology of refractory epilepsy in children]. AB - INTRODUCTION AND OBJECTIVES: The pathological findings in surgical material from children with refractory epilepsy has not offered yet a clear understanding of its role in this condition. The objective of this paper is to report our findings to further expand our knowledge about refractory epilepsy in children. MATERIAL AND METHODS: Results of microscopic examination of the surgical specimen obtained from 80 children, ages 12 or younger, who had surgery for intractable epilepsy at Miami Children's Hospital between 1990 and 1996 were reviewed. RESULTS: Examination was normal only in one. The rest revealed ectopic neurons (1), dysplastic cells with ectopic neurons (2), dyslamination with large neurons (7), dyslamination with ectopic neurons (18), dyslamination with dysplastic cells (10), pachygyria (2), encephalomalacia (9), gliosis with ectopic neurons (10), gliosis without ectopic neurons (3), developmental ectodermal tumor (6), ganglioglioma (2), tumors (3), and Rasmussen encephalitis (4). Lesions were located to the temporal lobe in 34 children. CONCLUSIONS. Extratemporal lesions are more frequent than temporal one, including hyppocampal sclerosis. Ectopic neurons, the most frequent pathological findings, rather than a cause of seizure may be a marker other highly epileptogenic cortical malformations. PMID- 9206606 TI - [The genetic diagnosis of children with mental retardation]. AB - Progress in cytogenetics and in molecular genetics has made it possible to apply these techniques to many aspects of medicine, to discover the pathogenesis of illnesses previously of unknown aetiology and to improve diagnosis, prognosis and prevention. They are also used in the preclinical diagnosis of individuals at risk, to identify the carriers of many genetic conditions and in prenatal diagnosis. In the analysis of mental retardation in children they are extremely useful, since they permit very early diagnosis. PMID- 9206608 TI - Ask the experts. The differential diagnosis of contact chemical leukoderma. PMID- 9206607 TI - [Neurometabolic diagnosis of mental retardation]. AB - Eighty-five percent of mentally retarded people have mild mental retardation (MR). Severe MR has a probable or definitive causes in 80% of cases. 2/3 are prenatal, 1/6 perinatal and 1/6 postnatal. Metabolic disorders may manifest as MR and its progressive nature often is not observed at the time of evaluation. MR is a prominent finding of many hereditary metabolic diseases, but only a fraction of MR seen in daily practice is due to inborn errors of metabolism. Medical conditions may present as MR without major extraneurological changes. In the investigation of MR, developmental milestones of affected children as well as detailed physical and neurological exam looking for physical characteristics, and performance of routine and special screening tests should be done. The metabolic screening tests (urine and/or blood) is very helpful. Use of tandem MS or acylcarnitine profile in blood for the diagnosis of organic acidurias and aminoacidurias can be done in 3 minutes per blood sample, with more than 20 conditions simultaneously tested. Blood spots in filter paper, once dried, are stable for weeks and can be sent by regular air mail, with low processing costs. Its future as a mass screening tool seems promising. PMID- 9206609 TI - Ask the experts. Patch testing for chemical leukoderma. PMID- 9206610 TI - Ask the experts. The role of ocular disturbances in the differentiation of idiopathic vitiligo from contact leukoderma. PMID- 9206611 TI - Selected highlights from the Jahassohn Centenary Congress. October 9-12, 1996. London, UK. PMID- 9206612 TI - The joy of discovery. PMID- 9206613 TI - Nuclear Cardiology and Managed Care: From Challenge to Opportunity. Proceedings of a symposium. Orlando, Florida, March 23, 1996. PMID- 9206614 TI - [Orthotopic neobladder in women]. AB - In women undergoing radical cystectomy for bladder cancer, orthotopic bladder reconstruction is now a viable alternative to urinary diversion: preservation of the external urethral sphincter by sectioning the urethra 0.5-1 cm distally to the bladder neck allows maintenance of urinary continence without compromising cancer control. 12 cases of bladder reconstruction in women operated on from 1986 to 1995 are presented here. A personal technique for the creation of an ileal neobladder is described: the use of staplers for detubularization of the ileum significantly reduces the operating time ("simplified ileal bladder"). Other important points of technique are as follows: 1. Careful preparation of the bladder neck and proximal urethra, staying above the pubo-urethral ligaments that must be preserved as the distal landmark of dissection; 2. "Nerve-sparing" isolation of the posterolateral wall of the bladder from the vagina; 3. Careful positioning of the pouch in the true pelvis, in order to avoid posterior prolapse of the neobladder. So far, results of bladder reconstruction in this series of patients are encouraging, both from a functional and oncological standpoint. PMID- 9206615 TI - [Electrovaporization of the prostate]. AB - Electrovaporization is a new and potentially useful modification of transurethral resection of the prostate which is rapidly becoming a treatment of choice for symptomatic benign prostatic hyperplasia. We report our experience with 22 patients treated using the Circon Acmi Grooved Vaportrode vaporization electrode. Procedures were carried out using the standard technique of electrovaporization with general or regional anaesthesia. Mean operative time was 40 minutes (range 25-65 minutes) and all patients were discharged without catheter after 1 or 2 days. Symptom score, peak uroflow and postvoid residual urine showed statistically significant improvement at 3 months (p < 0.01). There were no significant complications. Transurethral electrovaporization of the prostate seems to be a easy and safe procedure with significantly lower costs and excellent results. However, long-term efficacy and complications need to be evaluated in multicentre clinical trials. PMID- 9206616 TI - [Laparoscopic surgery of stress urinary incontinence in women]. AB - The Authors carried out a literature review on stress urinary incontinence (SUI) treatment using laparoscopy or pelvioscopy. They report their experiences in using a pubo-vaginal percutaneous colposuspension assisted by pelvioscopy. Of 22 treated patients, there was a total correction of the incontinence in 15 patients. (68.12%) with a median follow-up of 18.6 months. They report also their first experience with the Retzius plasty of Manhes by laparoscopy (3/4 patients treated are continent, 1/4 has recurrent SUI during times of stress). They conclude that laparoscopy and pelvioscopy treatment can be valid alternatives to traditional surgery if made on selected patients. PMID- 9206617 TI - [Anterograde transpelvic endopyelotomy]. AB - The Authors describe their own technique of antegrade endopyelotomy for cases of primary ureteropelvic junction obstruction. After percutaneous access has been gained via the lower calix, the technique involves making a wide opening of the renal pelvis and exploring the peripelvic space in front of a 3 to 4 cm long section of the ureter. The aim is to carry out the operation with an unrestricted view and without the need for a large calibre stent, in order to perform endopyelotomy in pediatric patients or in the presence of anomalous vessels. The follow-up demonstrates good results in 80% of 46 patients aged 5 to 62 years. The Authors think that antegrade transpelvic endopyelotomy is an endourologic operation with results and feasibility similar to those reported for open surgery. PMID- 9206619 TI - [Clinical competence of the surgeon and surgical risk. Teupitz Discussion 18 and 19 October in Motzen]. PMID- 9206618 TI - [Radical transcoccygeal prostatectomy]. AB - OBJECTIVE: A non randomized prospective study aimed at verifying the clinical outcome and pathological features of a group of patients submitted to transcoccygeal radical prostatectomy. METHODS: Radical transcoccygeal prostatectomy was carried out at our institution in 26 patients after laparoscopic (24 cases) or open surgical (2 cases) pelvic lymphadenectomy. Eighteen patients were selected if considered at risk for nodal metastases on the basis of preoperative staging (PSA > or = 20 ng/ml and/or Gleason score > 5), while the remaining 8 were affected by incidental prostate carcinoma. RESULTS: Intraoperative complications included rectal injury and massive blood los in one case (3.8%). Transitory leakage at the site of the urethrovesical anastomosis and urethrorectal fistula occurred postoperatively in two patients. The rate of positive surgical margins was 26.9%. The mean follow-up time was 27 months (range 3-39 months). Total urinary continence was obtained in 21 cases (80.8%), while 5 patients (19.2%) still require urinary pads. Four patients (15.4%) have experienced tumour recurrence evidenced only by elevated serum PSA levels. Local tumour recurrence with positive biopsy of urethrovesical junction was diagnosed in 3 patients (11.5%), while systemic tumour recurrence occurred in one case (3.8%). CONCLUSIONS: Radical transcoccygeal prostatectomy is a safe procedure for the surgical treatment of prostate cancer both from a clinical and pathological point of view. Operative complications, as well as pathological features and clinical outcome reported in this series of patients, must be related to selection criteria use in the majority of cases. The exact role of radical transcoccygeal prostatectomy in the clinical setting has yet to be defined. According to these preliminary results, radical transcoccygeal prostatectomy should be further investigated in the treatment of incidental carcinoma following TURP or suprapubic prostatectomy. PMID- 9206620 TI - [Professional liability insurance]. PMID- 9206621 TI - [Efficiency and economy in surgery]. PMID- 9206622 TI - [Quality of a periodical]. PMID- 9206623 TI - [Development and pathophysiology of abdominal wall defects]. AB - Repair of abdominal wall hernias is the most frequently performed operation in surgery. Primary hernias arise at anatomical weak points; incisional hernias result from laparotomies. As the fascia heals slowly, the suture should be strong enough to withstand the intraabdominal pressure. Complications in wound healing may lead to a hernia, especially when absorbable sutures have been used. The rate of incisional hernias is technically influenced by the type of incision, the suture technique, and the suture material. To achieve adequate repair it is often necessary to change the principles of abdominal wall closure, for example by using alloplastic material. The meshes reinforce the architecture of the abdominal wall both by pure mechanical means and by induction of a stable scar formation (polypropylene, polyester). PMID- 9206624 TI - [Incisional hernias]. AB - Incisional hernias are a common complication following abdominal surgery. Surgical treatment as a whole has the disadvantage of having a high recurrence rate. Far better results are obtained by tension-free closure, if necessary backed up by a synthetic prosthesis (PTFE, polypropylene). However, even then, a recurrence rate of about 10% can be expected. PMID- 9206625 TI - [Tension-free suture of incisional hernia]. AB - Incisional hernias are a common complication following abdominal surgery. However, when performed correctly, the techniques utilized for direct hernia repair, Mayo's closure and plastic covering of the defect appear to be effective in the management of primary hernia closure. The recurrence rates of 30-50% continue to represent a significant challenge in the initial management of incisional hernia repair. Fortunately, there is evidence that, with correct preparation of the patient and with correct anatomical reconstruction, better results can be achieved. Also, the use of strictly applied technical skills in combination with standardized operative techniques may help to eliminate further complications and recurrences. In addition, the application of mesh or cutis transplantation to reinforce the attenuated tissue has also been shown to reduce the recurrence rate. With the diligent application of the above measures, as well as of new concepts, which at present include a promising technique called tension adapted closure, there may be positive developments in the treatment and management of incisional hernias. PMID- 9206626 TI - [Surgical relevance of preoperative diagnosis of tumors of the gastrointestinal tract--decision making in esophageal, stomach, colon and rectal carcinoma]. AB - The increasing spectrum of therapeutic options for tumors of the gastrointestinal tract has resulted in a refinement of the pretherapeutic diagnostic strategies. The diagnostic approach in surgical institutions that are focused on primary surgical resection will therefore be much less sophisticated than in institutions who propose a selective therapeutic approach based on the pretherapeutic tumor stage and prognostic parameters. Pretherapeutic assessment of the depth of tumor infiltration, i.e. the T-category, is essential because most further diagnostic and therapeutic decisions are based on this information. This can today be achieved with a high degree of accuracy by endoscopy and endoscopic ultrasonography. Early T-stages (T1-2) are usually an indication for primary surgical resection and, after exclusion of distant metastases, no further diagnostic studies are required. In patients with locally advanced esophageal, gastric or rectum tumors (T3-4) multimodal therapeutic concepts should be considered. This usually requires additional diagnostic studies. None of the available diagnostic imaging modalities today allows satisfactory pretherapeutic assessment of lymph node metastases. The assumed nodular status should therefore currently not influence therapeutic decisions. Essential is, however, the assessment of distant metastases, since the documentation of distant tumor spread will change the therapeutic approach to a palliative situation. Detailed histologic and molecular-biologic assessment of tumor characteristics is growing in importance. This not only provides therapeutically relevant information regarding tumor grading, but opens the door towards a modern molecular diagnostic approach. It can be expected that in the near future a vast amount of relevant prognostic information can be obtained from endoscopic tumor biopsies, which may soon alter our therapeutic concepts. PMID- 9206627 TI - [Surgical relevance of diagnostic imaging of abdominal tumors--decision making in pancreatic tumor]. AB - Imaging of tumors of the pancreas may have a series of significant implications for surgical decision making. First of all verification and localization of a suspected tumor is crucial. Later on, accurate staging of local tumor extent and of distant metastases is necessary for evaluating the indications for surgical intervention. If histologic proof of malignancy of a pancreatic lesion is needed, different imaging techniques can be used for percutaneous biopsy of the tumor. Finally imaging is required in patients with a pancreatic tumor if an obstruction of the common bile duct is treated with palliative intent by endoscopy or transhepatic intervention. A rational and efficient selection from all the imaging techniques currently available requires a clear-cut definition of what is needed for surgical decision making in each individual case. Thereby one has to take into account whether the tumor arises from the exocrine or from the endocrine tissue of the pancreas. PMID- 9206628 TI - [Significance of diagnostic imaging for determining surgical indications in solid liver tumors]. AB - Radiological imaging is essential for the differential-diagnostic appraisal and the assessment of size, number, and localization of neoplastic liver lesions. The results determine the decision for or against operation and the surgical strategy. Moreover, oncological contraindications against surgery are recognized by imaging procedures. Technical refinements of imaging comprise improved quality of ultrasound and of computed tomography and fast sequences in magnetic resonance imaging. Special investigational protocols and specific contrast agents for ultrasonography and magnetic resonance technique are highly relevant for clinical practice. The value of diagnostic procedures is demonstrated by our own experience. The differential use of imaging procedures, technical and methodological performance and the final diagnostic interpretation require a high standard of expertise. Diagnostic accuracy is limited by technical and biological factors. PMID- 9206629 TI - [surgical relevance of diagnostic imaging in abdominal tumors--decision making in retroperitoneal tumors]. AB - The indication for surgical treatment of tumors of the retroperitoneum should not depend on findings of radiological examinations, because all radiographic procedures lack tissue specificity. Instead, it can only be determined histologically after an open biopsy or resection of the tumor, which should always be attempted. With the help of appropriate radiological examinations, a retroperitoneal mass will not only be detected, but the relationship to the adjacent anatomical structures will also be delineated. Therefore, valid information for planning the operation and estimating the morbidity and mortality can be provided for the surgeon. PMID- 9206630 TI - [Duodenum preserving resection of the head of the pancreas: a standard procedure in chronic pancreatitis]. AB - Duodenum-preserving resection of the head of the pancreas was developed 25 years ago by Beger. This procedure is indicated in patients suffering from chronic pain in combination with inflammation of the head of the pancreas, common bile duct obstruction, pancreatic duct obstruction and/or obstruction of the retropancreatic vessels. At the Inselspital in Berne, 74 patients underwent this operation between 1993 and 1996. The median length of the operation was 380 min, with the need for transfusion in a median of 0 units (0-6). There was no postoperative mortality. Total postoperative morbidity was 13%. One patient needed relaparotomy on day 17 for small bowel obstruction. Median length of hospital stay was 11 days. Postoperatively, two patients developed diabetes. Duodenum-preserving resection of the head of the pancreas represents an organ preserving principle of surgery. This procedure treats the complications of chronic pancreatitis and provides long-term pain relief in more than 80% of patients. PMID- 9206631 TI - [Drainage versus resection in surgical therapy of chronic pancreatitis of the head of the pancreas: a randomized study]. AB - Drainage and resection are the principles of surgery in chronic pancreatitis. The techniques of duodenum-preserving resection of the head of the pancreas as described by Beger and Frey combine both to different degrees. In a prospective randomized trial both procedures were compared: 74 patients were randomly allocated to either Beger's (n = 38) or Frey's, (n = 36) group. In addition to routine pancreatic diagnostic work-up a multidimensional psychometric quality-of life questionnaire and a pain score were used. Assessment of endocrine and exocrine function included oral glucose tolerance test, serum concentrations of insulin, C-peptide, and HbA1c, as well as fecal chymotrypsin and pancreolauryl test. The mean interval between symptoms and surgery was 5.1 years (1-12 years). The median follow-up was 30 months. There was no mortality. Overall morbidity was 27% (32% Beger, 22% Frey). Complications from adjacent organs were definitively resolved in 91% (92% Beger, 91% Frey). A decrease in pain score of 95% and 93% after Beger's and Frey's procedure, respectively, and an increase of 67% in the overall quality-of-life index in both groups were observed. Endocrine and exocrine function did not differ between the two groups. Both techniques of duodenum-preserving resection of the head of the pancreas are equally safe and effective with regard to pain relief, improvement of quality of life, and control of complications affecting adjacent organs. Neither procedure leads to further deterioration of endocrine and exocrine pancreatic function. PMID- 9206632 TI - [Malignant tumors of the hepatic bifurcation--results of surgical therapy and prognostic factors]. AB - Between 1 January 1986 and 31 December 1994, 45 patients with carcinoma of the hepatic bifurcation underwent operations in the department of surgery. According to Bismuth's classification (1992), there were 7 type I, 4 type II, 22 type III and 12 type IV tumors. 31 of the 45 patients (68.9%) underwent surgical resections with curative intentions: resection of the hepatic bifurcation (n = 9, 29%), resection of the hepatic bifurcation with hilar liver resection (n = 7, 22.6%), resection of the hepatic bifurcation with left hemihepatectomy (n = 10, 32.3%), orthotopic liver transplantation (n = 5, 16.1%). The remaining 14 patients underwent palliative (n = 6) or diagnostic (n = 8) operations. Two of the 45 patients (4.2%) died in hospital. Histological examination demonstrated R0 resection in 17 of 31 (54.8%) of the patients, while in the remaining 14 cases, only a R1/RX resection had been achieved. One-, 2- and 5-year survival rates were 75.4%, 53.9% and 24.5% (median survival: 729 days) in the 31 resected patients, and 30.8%, 10.3% and 0% (median survival: 153 days) in the palliative treatment group. A statistically significant influence on survival could be demonstrated for nodal status (N0/N1) and residual tumor status (R0/R1, R2). PMID- 9206633 TI - [Therapy of persistent bleeding esophageal varices using intrahepatic portosystemic stent shunt and immediate liver transplantation]. AB - A 41-year-old patient with liver cirrhosis due to autoimmune hepatitis received an emergency transjugular portosystemic stent shunt for uncontrolled acute variceal hemorrhage. Because of markedly impaired liver function, liver transplantation was considered to be indicated and was performed on the following day. Intraoperatively, one of the intrahepatic metal stents migrated unnoticed into the pulmonary artery. The postoperative course was uncomplicated and the displaced stent was left in situ. Eighteen months after the transplantation the patient is well with normal liver function and no pulmonary problems. PMID- 9206634 TI - [Surgical therapy of esophageal perforation. A determination of current status based on 4 personal cases and the literature]. AB - We present four cases of esophageal rupture (three iatrogenic, one Boerhaave syndrome) to demonstrate the difficulty in diagnosis and therapy. The current literature is discussed and conclusions are drawn as regards the modus operandi. All our patients were operated on. The site of esophageal rupture was always closed with a primary suture and substantial irrigation and drainage were performed. In two cases the suture line was in addition covered with fibrin glue or by an omentum flap, respectively. All patients survived and recovered was unremarkable. Our own results and a subsequent analysis of literature allow the following conclusions. At an early stage of esophageal rupture surgical intervention is indicated. The method of choice is primary closure of the rupture site by suture, possibly combined with a muscle or omentum flap. In cases of delayed diagnosis with advanced mediastinitis, suture of the rupture site should also be striven for. Additional coverage is advisable in these cases. Resection procedures with or without reconstruction should be done only in exceptional cases before of the high surgical risk. PMID- 9206635 TI - [Forensic aspects of complicated laparoscopic cholecystectomy]. AB - As laparoscopic cholecystectomy becomes more established, interest in legal aspects increases. The legal cases dealt with in Germany since the introduction of this laparoscopic method in 1989 were studied to determine the legal consequences of complications. Of 40 cases, 16 were judged to be cases of malpractice, whereas 24 were not. The most common cause was injury of the common bile duct (26 cases). Seven of these 26 cases were judged to be cases of malpractice, 2 of these as grave errors. The main reasons for the acceptance of a case as malpractice were delay in changing to conventional cholecystectomy, delayed revision, laparoscopic revisions and not reverting to conventional cholecystectomy in unclear situations. PMID- 9206636 TI - [Aortic aneurysm in horseshoe kidney--special diagnosis and therapy]. AB - The co-existence of horseshoe kidney (HSK) and abdominal aortic aneurysm (AAA) is rare and demands a special diagnostic workup and a meticulous surgical procedure. Ten patients with HSK associated with AAA are reported. All underwent aortic replacement with successful preservation of multiple renal arteries. One patient died at the 7th postoperative day from myocardial infarction. HSK does not represent a contraindication for aortic repair. Angiography for identification of aberrant renal arteries and classification into three types for surgical management is mandatory. In type I and II we recommend a standard midline transperitoneal approach. Only in type III does a thoraco-abdominal approach seem favourable. The aortic reconstruction should be performed with tube grafts, if possible. Aberrant renal arteries are reattached directly to the prosthesis. Dissection of renal isthmus should be avoided. Temporary cold renal perfusion is indicated to extend the ischemic tolerance time. As repair of a ruptured AAA in patients with HSK may be quite difficult and time consuming, we recommend more liberal indications for aneurysm surgery in patients with HSK. PMID- 9206637 TI - [Prolonged intensive care stay after abdominal surgery interventions with special reference to quality of life, occupational rehabilitation and economics]. AB - Owing to increasing limitations on resources in health care, there is an urgent need to investigate effectiveness and efficiency of medical procedures. Therefore, we retrospectively studied the courses of 62 surgical patients who required at least 30 days of intensive care regarding mortality, long-term prognosis and quality of life. Additionally, a cost analysis was made using quality-adjusted life years (QALYs). The hospital mortality was 40.3%. The overall median survival time of discharged patients (n = 37) was 3.7 years and the calculated 3-year survival was 56.4%. The most frequent causes of death were septic complications or multiple organ failure in hospitalized patients and tumor relapses in discharged patients. In most of the surviving patients quality of life (median Gastrointestinal Quality of Life Index: 104 points) was good. About 20% of the discharged patients were able to return to work. Although extended intensive care therapy is extremely expensive (DM 68,250 per QALY), these costs are comparable with other accepted procedures in medicine (i.e. hemodialysis). Therefore, economical aspects should not be a generalized reason for withdrawing or withholding intensive care therapy. PMID- 9206638 TI - [Cost savings by disinfection for prevention of surgical wound dehiscence after gastrectomy]. AB - The aim of this study was to examine the effect of decontamination as compared to placebo medication on post-gastrectomy treatment costs. The results of a prospective double-blind placebo-controlled multicenter trial indicate that perioperative i.v. prophylaxis with cefotaxim and topical decontamination with polymyxin B, tobramycin, vancomycin and amphotericin B from the day before surgery until the 7th postoperative day is most effective in the prevention of esophagojejunal anastomotic leakage following total gastrectomy. For the cost analysis, only patients who had been decontaminated according to the study protocol (n = 90) were compared to the non-decontaminated patients (n = 103). The esophagojejunal leakage rate was 10.6% in placebo patients (n = 103) and could be reduced significantly to 1.1% in decontaminated patients (n = 90, P = 0.0061; two tailed Fisher's exact test). There was only one asymptomatic leakage detected on Gastrografin swallow. The pulmonary infection (P = 0.0173) and overall complication rates (p = 0.0238) were significantly reduced in the decontamination group as well. During the observation period, 9 (8.7%) patients in the placebo group and 3 (3.3%) in the decontaminated group died (P = n.s.). Patients were followed up for the initial 42 postoperative days and treatment costs were calculated for this time period only. The parameters compiled in the study pertaining to use of medical resources formed the basis for the determination of the postoperative treatment costs. These were the costs for decontaminating drugs, intravenous antibiotics, reoperations and non-surgical reinterventions as well as daily treatment costs of the general ward, the intensive care unit (ICU) and rehabilitation. The average costs per patient in the placebo group amounted to DM 20,000 while the costs for decontaminated patients were only DM 16,200, which was due to a significantly lower number of patients requiring treatment in the ICU (P = 0.0082), significantly fewer patients requiring i.v. antibiotics (P = 0.0232) and fewer patients with reoperations (P = 0.0909). The prophylaxis employing decontaminating drugs in the amount of DM 400 lowered post-gastrectomy treatment costs by DM 3800 or 19%. The prophylaxis can be recommended, because it lowers morbidity, mortality and the costs of total gastrectomy. PMID- 9206639 TI - [Posterior resection of of the head of the pancreas in malignant duodenal gastrinoma]. AB - We report on a case of malignant gastrinoma located on the posterior surface of the descending duodenum, presenting with Zollinger-Ellison syndrome. The tumor was not evident on preoperative imaging studies, metastasis was not present and there was no coincidence with multiple endocrine neoplasia type-I. As the gastrinoma was located on the posterior surface of the pancreatic head, to obtain a sufficient safety margin, partial excision of this region was necessary. Under preservation of the Oddi's sphincter, the reconstruction was completed by direct suturing of the duodenal wall to the pancreatic surface without need for enteral diversion procedures. This technique represents a possible non-invasive resection modality for benign and malignant duodenal gastrinomas located close to the pancreatic head region. PMID- 9206640 TI - [Simultaneous retroperitoneal operation of juxtarenal abdominal aortic aneurysm and ischemic vertebral body necrosis]. AB - The combination of abdominal aortic aneurysm (AAA) and necrosis of the lumbar vertebral bodies is often the consequence of ischemia of the lumbar arteries and local compression from the aneurysm. A patient with necrosis of lumbar vertebral bodies 2 to 4 was admitted for abdominal aneurysm repair. CT scanning revealed almost complete destruction of the second and fourth lumbar vertebral bodies. In a combined operation an orthopedic and a vascular surgical team implanted two carbonic cages with autogenous splinter of the pelvic bone and an aortic vascular graft, using a retroperitoneal approach. Three months after the operation the 61 year-old man is entirely well and without any signs of back pain. He could be fully mobilized within 3 weeks postoperatively. This case study depicts the surgical techniques and discusses the advantages of the simultaneous operation and retroperitoneal exposure. PMID- 9206641 TI - [Computer-based training--a new method in surgical education and continuing education]. AB - Computer-based training (CBT) programs teach the material of a specific field and at the same time offer various ways of objectively checking the knowledge gained. The interactive use of multimedia components, such as text, graphics, animation, sound, digital slide shows, videos and quizzes, facilitates the learning process. The aim of this study was the development and evaluation of a CBT program for use by surgeons teaching students. Using SuperCard, a teaching module for distal radius fracture (DRF) was developed, containing detailed clinical information. Video clips and vivid animation combine theoretical knowledge with practical experience. Fourth-year medical students (n = 103) were tested after using the module for 90 min. Other students (n = 47) served as the control group. In a 90 min lecture, DRF was discussed. In all evaluated criteria (distinctness, detailed description, presentation of materials, structure, motivation to learn, time saved while learning and memory retention), CBT gained 15-20% better scores than the lecture. Although 87% of the students stated that their experience with computers was limited or insufficient, 100% found the use of CBT systems helpful in student teaching. Most of them suggested the use of such programs as a exam preparation/self study method (90%) or as a supplement to a lecture (40%). Based on these evaluations, it is clear that CBT modules are an appropriate future teaching and learning system that will be well accepted. In conclusion, CBT programs should be integrated into medical education as a valuable supplement. With this aim, CBT systems should be developed and used at universities as an information system for the surgical residency program. PMID- 9206643 TI - [Removal of endoscopic percutaneously implanted gastrostomy tubes. Comment on the contribution by H. Seitz et al]. PMID- 9206642 TI - [Analysis of the publication spectrum of 4 German-language medical specialty periodicals "Der Chirurg", "Der Unfallchirurg", "Langenbecks Archiv fur Chirurgie" and "Medizinische Klinik"]. AB - All 1994 publications of four leading journals in surgery, orthopedics and internal medicine in Germany (Der Chirurg, Der Unfallchirurg, Langenbecks Archiv fur Chirurgie, Medizinische Klinik) were reviewed by means of a structured analysis. The type of article, authorship, number of references, geographical location, quality, and main conclusion (positive; neutral; negative) of each article were documented. The journals focused on clinical studies (32.2-59.6%), case reports (11.0-26.1%) and reviews (6.4-40.9%). Articles about surgical techniques were mainly found in Der Chirurg (16.9%), experimental studies in Der Unfallchirurg (14.7%) and Langenbecks Archiv fur Chirurgie (25.7%). Most articles were written by university clinic personnel (62.6-81.4%). In 11.0-22.6% of all articles, the head of a clinic was the first author and the co-author in 36.5 58.7%. Women were found to be first author in 4% in surgery, and in 10.4% in internal medicine. Of a total of 495 publications, 53.9% were written in Northrhine-Westphalia, Bavaria or Baden-Wurttemberg and only 1.6% in the five new federal states. Of all articles 16.4% were by foreign authors, with 10.5% originating from German-speaking countries. The portion of controlled randomized trials ranged between 4.9% (Der Unfallchirurg) and 11.3% (Der Chirurg) of all published studies. Studies and case reports with negative results were found to be more evident in Der Chirurg and Langenbecks Archiv fur Chirurgie (ca. 20%) as compared to Der Unfallchirurg and Medizinische Klinik (ca. 6%). All journals provided good general information about the actual developments in a variety of topics to the reader. However, some improvement concerning international contributions, participation of the new federal states, and the quality of studies is recommended. PMID- 9206644 TI - [Endoscopic hernia surgery (TAPP)--gold standard in management of recurrent hernias? Comment on the contribution by B. Leibl et al]. PMID- 9206645 TI - [Endoscopic hernia surgery (TAPP)--gold standard in the management of recurrent hernias? Comments on the contribution by B. Leibl et al]. PMID- 9206646 TI - [Endoscopic hernia surgery (TAPP)--gold standard in the management of recurrent hernia? Comments on the contribution by B. Leible et al]. PMID- 9206647 TI - [Possible HIV infection: when exposure prophylaxis?]. PMID- 9206648 TI - [Helicobacter pylori--talked about by both patients and physicians]. PMID- 9206649 TI - [Helicobacter pylori--when and how do gastroenterologists treat themselves? A clinical and practical survey]. AB - OBJECTIVE: To determine how often gastroenterologists know their own Helicobacter pylori status, how they diagnose the infection and what consequences they draw from the findings. METHODS: Between November 1995 and March 1996 questionnaires were sent to gastroenterologists in hospitals (592, with five not reached) or private practice in Germany (431, 12 not reached), with a response rate of 24.4% and 55.4%, respectively. RESULTS: Of the 375 respondents 27 were female; the average age was 47.6 +/- 7.8 years. 151 knew their own H. pylori status (40.3%), with an infection rate of 55.6%. Most of the doctors had used noninvasive diagnostic tests (serology: 18.5%, breath test: 37.7%, serology + breath test 2.0%). 54 doctors (among them three without prior testing) had treated themselves between 1992 and 1995, ulcer or dyspepsia having been the most common indication (in 60%). Between 1992 and 1994 omeprazole plus amoxycillin was the most common form of treatment, while in 1995 a triple regimen with omeprazole and two antibiotics was most common during 1995. Either regimen was well tolerated in 85% of those treated, but about a quarter of doctors reported difficulties in compliance. Only half of the participants tested the result of treatment, similar numbers using either the breath test (n = 14) or biopsy (n = 13) for confirmation. CONCLUSIONS: A significant percentage of gastroenterologists in Germany knew their own H. pylori status. However, the diagnostic procedures and indications for treatment often did not respond to recommended guidelines. The frequency of treatment and the regimen used mirrored the improved knowledge of the significance of H. pylori infection and the development of antibacterial treatment during the last few years. PMID- 9206650 TI - [Complicated intestinal tuberculosis of the mesenteric root]. AB - HISTORY AND CLINICAL FINDINGS: In the course of 2 1/2 years a 72-year-old man had five episodes of intestinal bleeding with anaemia, but no cause was at first found by routine diagnostic tests. After the fourth episode a diagnostic laparotomy was performed at which a diffusely infiltrating growing tumour was found. As it encircled the mesenteric vascular axis resection was not possible. But because of threatened ileal obstruction by the tumour a segmental ileal resection was performed, but no intraoperative diagnosis could be established. INVESTIGATIONS: Repeated oesophago-gastro-duodenoscopies and a coloscopy all revealed blood, especially in the upper gastrointestinal tract, but no bleeding source was found. Angiography suggested bleeding in the terminal ileum. Computed tomography demonstrated a tumour in the region of the mesenteric root. DIAGNOSIS, TREATMENT AND COURSE: At a second laparotomy because of another bleeding, which required a blood transfusion, tuberculosis was diagnosed. Triple drug treatment was initiated and the patient has been free of symptoms and bleedings since then and fully active for 1 1/2 years. CONCLUSION: This case clearly demonstrates the difficulty of diagnosis and the long delay until specific treatment, because the combination of "tumour", bleeding and stenosis of the small intestine by an infiltrative lesion at first pointed to malignant process. PMID- 9206651 TI - [The curative therapy of primary hyperparathyroidism by percutaneous ethanol injections]. AB - HISTORY AND CLINICAL FINDINGS: An 86-year-old woman was admitted because of acute nonspecific upper abdominal symptoms and vomiting. She was occasionally disoriented, generally slower in movement and reaction, apathetic and mainly bed ridden. She was a known insulin-dependent diabetic who had sustained a posterior wall myocardial infarction and cerebrovascular accident and had undergone a cholecystectomy. On physical examination her upper abdomen was painful to pressure, blood pressure was 180/95 mm Hg, but there were no other findings. INVESTIGATIONS: Sonography demonstrated bile-duct dilatation, confirmed at endoscopic retrograde cholangiopancreatography, and a prepapillary choledochal concrement of about 10 mm. Sonography also revealed an echo-poor tumour of the right caudal parathyroid. The calcium concentration was raised to 2.94-3.16 mmol/l and the parathormone level was also increased (99.5 pmol/l, normal 1.2-5.7 pmol/l), as were amylase (375.6 U/l) and lipase (1038-5394 U/l). TREATMENT AND COURSE: After papillotomy and extraction of the choledochal concrement the acute biliary pancreatitis quickly improved. Operation on the parathyroid tumour was not undertaken because of the patient's various illnesses. Instead, 95% alcohol was instilled, 3.5 and 4.5 ml respectively, into the tumour, under sonographic control in two sessions, 3 days apart. Her clinical condition clearly improved and serum calcium became normal and the parathormone level fell significantly. CONCLUSION: Percutaneous ethanol injection of a parathyroid tumour can be a curative and sparing alternative to operation in patients with hyperparathyroidism seemed too ill for surgery. PMID- 9206652 TI - [Obliteration of the large arteries as a late sequela of radiation-induced vasculopathies]. PMID- 9206653 TI - [Sleep, breathing and gastroesophageal reflux]. PMID- 9206654 TI - [Hyperthermia]. PMID- 9206655 TI - [Angina pectoris with normal coronary arteries]. PMID- 9206656 TI - [The heterotopic ossification of a Dacron prosthesis in peripheral arterial occlusive disease]. PMID- 9206657 TI - [Psychiatric and psychologic aspects of stomatologic diseases, or stomatologic aspects of psychiatric diseases. Review of the literature]. AB - Dental symptoms, oral medicine and psychiatric and psychologic problems have strict relationships in several domains. Behavior and behavioural problems (deficient oral hygiene, lack of regular dental control, dependence of nicotine or alcohol, etc.) as well as certain psychiatric diseases influence the patients' dental state to a great extent. There are further problems determined by the different types of anxiety, fear and bad previous experiences which have an impact on people's attitudes towards dental treatment and the development of hygienic habits. Dentists' psychologic and psychiatric knowledge can have a considerable contribution to the reduction of the patients' anxiety, furthermore to an appropriate treatment, by the recognition of the underlying psychiatric disease. PMID- 9206658 TI - [Use of drugs in stomatology II. Use of antibiotics in dental practice]. AB - The authors summarized 514 questionner on dental patient medication, and established that, the dentists use an average in Hungary 29 box antibiotics per months. The most frequently prescribed medicine (Dalacin C, Rulid, Augmentin, Semicillin, Maripen, Doxycycline) take the 75% of the total number. The 52% of dentists use antibiotics for prophylactic aims, 94.6% for the treatment of inflammatory diseases. The examination gave data for creating the picture of the use of antibiotics in Hungarian dental practice in 1995. PMID- 9206659 TI - [Mutation "outbursts" of various genes in natural populations of Drosophila melanogaster]. AB - Mutation outbursts of several sex-linked genes are described. The outbursts occurring in natural populations of Drosophila melanogaster in various periods are shown to result mainly from transpositions of mobile genetic elements. PMID- 9206660 TI - [Structural features of the modified BARE-retroelement in the barley (Hordeum vulgare L.) genome]. AB - The primary structure of the 4.2-kb BamHI-fragment occurring abundantly in the genome of barley Hordeum vulgare was determined. By means of computer analysis, considerable homology was found between this fragment and the copia-like BARE-l retrotransposon studied earlier. A unique distinction of the BamHI fragment is its symmetrical structure caused by the presence of two mutually inverted parts, each of which is homologous to a long region of BARE-l including a 5'-LTR (long terminal repeat) and the adjacent leader sequence. No sequences homologous to the coding domains of BARE-l were revealed in the fragment. However, potentially functional signals were found: TATA boxes and primer-binding sites (PBS) exhibiting statistically significant homology to the corresponding regulatory signals of known retroelements. Thus, we have revealed and characterized a repeated element of the H. vulgare genome that is a markedly modified derivative of the BARE family of retrotransposons of this genome. PMID- 9206661 TI - ["SPELT1"--a new family of tandem repeats in grasses]. AB - A novel family of tandemly repeated DNA in the cereal genome, "Spelt1," was isolated and described. This family accounts for about 2% of the DNA of Aegilops speltoides, one of the putative donors of B genome of hexaploid wheat. Genomes of fifteen other cereal species studied, including those with AB, AG, and ABD genome formulas, contain significantly lower amounts of Spelt1 repeats. RFLP analysis revealed three different types of Spelt1 sequence organization in the wheat and aegilops genomes studied. Data indicating that Spelt1 is a specific marker of Ae. speltoides-chromosomes are presented. PMID- 9206662 TI - [Structural and functional chromatin organization of the SUP35 gene in Saccharomyces cerevisiae yeast]. AB - Structural and functional organization of the 5' region of the SUP35 gene was analyzed in Saccharomyces cerevisiae yeast. Indirect DNA end labeling allowed two nuclease-hypersensitive sites and a region involved in nucleosomes to be revealed. DNase I and micrococcal nuclease hypersensitive sites were localized to almost the same regions: -461 ... -372 bp and -271 ... -91 bp for DNase I and 461 ... -356 bp and -231 ... -79 for micrococcal nuclease. Nucleosomes were localized to a region +22 ... +339 bp. Both the location of DNase I and micrococcal nuclease hypersensitive sites within the promoter region and the location of nucleosomes within the coding region remain the same at different cell culture growth phases. However, positioning nucleosomes were revealed within the SUP35 coding region only at the late logarithmic phase; the radioautographic pattern of them does not depend on the extent of nuclease digestion. PMID- 9206663 TI - [Genomic incompatibility in Drosophila virilis Sturt. X Drosophila lummei Hackman hybrids]. AB - Genetic analysis of male sterility in Drosophila virilis x D. lummei hybrids was performed. It was shown that hybrid male sterility was caused mainly by incompatibility of sex chromosomes X and Y and homozygous chromosome 2 of D. virilis with other autosomes of D. lummei. Genes of chromosomes 3 and 5 enhanced sterility whereas genes of chromosome 4 suppressed its development. Factors controlling hybrid male sterility were demonstrated to have a recessive type of inheritance. At 17 degrees C, chromosome 6 of D. lummei was eliminated in F1 hybrids of the cross female D. virilis x male D. lummei. At 17 degrees C and 25 degrees C, sterility of the F1 hybrid males was 96% and 9%, respectively. Thus, two isolation mechanisms-elimination of chromosome 6 and male sterility-can simultaneously operate in F1. In addition to the region of the species-specific inversion In 1(a+b), factors controlling male sterility were mapped to the vermillion region of the X chromosome. PMID- 9206664 TI - [Seasonal changes in the resistance of a Drosophila population to low temperatures and their association with fertility]. AB - Significant genetic variation in resistance to cold was detected in samples collected from a natural Drosophila population in spring, summer, and autumn. In the summer sample, phenotypic variation was determined by genetic factors; in the summer and autumn samples, exclusively by environmental factors. Flies collected in spring had the highest longevity at low temperatures. In summer, their longevity at low temperatures was drastically reduced but it was shown to increase in autumn. Cold-resistant flies had high fertility in spring and low fertility in autumn. The seasonal differences in resistance to cold were genetically determined and are probably caused by natural selection. PMID- 9206665 TI - [Interlocus associations of various genetic biochemical systems in cattle]. AB - Variation of some genetic biochemical systems (transferrin, ceruloplasmin, amylase-I, and kappa-casein) were studied in five cattle groups of different origin. Data on linkage disequilibrium demonstrated that some syntenic genes did not exhibit interlocus association. Conversely, interlocus association appeared between some loci that were located in different chromosomes. Apparently, the patterns of these associations may be specific for gene pools of individual groups from the same animal species. PMID- 9206666 TI - [Gradualism or punctualism: data on genetic differentiationof small mammals from the Holarctic region]. AB - Genetic differentiation of taxa from three Holarctic and three originally Afrotropical phyla of small mammals was analyzed. Its extrapolation to the time scale revealed the following trends: (1) Distributions of fixed gene differences along the scale are relatively independent. (2) Extrapolation of these distributions to the time scale shows that the speciation process is discontinuous. It consists of relatively stable periods interrupted by speciation bursts. (3) Periods of speciation activity in different phyla coincide. In general, these results are consistent with the concept of punctuated equilibria and suggest that the speciation process is temporally discontinuous. PMID- 9206667 TI - [Mutagenic effect of nitrosodimethylurea in male mice. Induction of dominant lethal mutations and chromosome aberrations in germ cells; genotype influence on the clastogenic effect in bone marrow cells]. AB - The ability of dimethylnitrosourea (DMNU) to induce dominant lethal mutations in germ cells and chromosome aberrations in bone marrow cells of male CBAB6F1 mice was studied. At a mutagen dose of 200 mg/kg, mortality was 19% in the second week after treatment (being 5% in the control) and the frequency of bone marrow cells containing chromosome aberrations was 19.6% 24 h after treatment. DMNU induces dominant lethals in postmeiotic cells and in spermatogonia; the effects in spermatozoids and spermatogonia are equal. Chromosome aberrations in spermatocytes induced by DMNU are not realized as dominant lethals. The sensitivity of mouse strains to the clastogenic effect of DMNU ranged in an order similar to that observed in experiments with thioTEPA. The most sensitive was the TPS strain (29.2 +/- 4.6% of cells with chromosome aberrations), the most resistant-the CBA/Lac strain (9.5 +/- 2.9%). DMNU exhibited a relatively poor clastogenic activity; the effect in bone marrow cells was higher than that in male germ cells. PMID- 9206668 TI - [Negative relationship between variation of nuclear and mitochondrial genomes in populations of Arctic Mongoloids of Northeastern Asia]. AB - Incompatibility of variation of nuclear and mitochondrial genomes was shown in four Arctic Mongoloid populations. An increase in heterozygosity was accompanied by a decrease in variation of mtDNA in the Koryaks-Chukchi-Asian Eskimo population series, and corresponds to the gradient of severity of the environment. An explanation is proposed based on the assumption of the key role of selective processes in the formation of the genetic (nuclear and mitochondrial) structure of Arctic Mongoloid populations in connection with long term adaptation to extreme environment. PMID- 9206669 TI - [Population-demographic structure of the population of Kursk district. Ethnic composition and age of marriage]. AB - When studying the marriage structure of the Kurskaya oblast population in 1987 1990, a positive marriage assortativeness with respect to ethnicity was found (K = 0.244). The Russian population exhibited panmixia; in the Ukrainian population, negative marriage assortativeness and panmixia prevailed; other ethnic groups were characterized by more or less pronounced positive marriage assortativeness. The ethnic compositions of raion (district) populations were determined by their geographic location and history. The parameter of marriage assortativeness with respect to age was r = 0.911; this value in the reproductive part of the population was r = 0.748. The age of contracting marriage in the raion populations depended on the extent of their urbanization. PMID- 9206670 TI - [Amplification of hypervariable genomic regions for establishment of the type of hematopoiesis in hemoblastosis patients after allogeneic bone marrow transplantation]. AB - To establish the type of hemopoiesis in 15 patients with hemoblastosis subjected to allogenic bone marrow transplantation, the amplification of four hypervariable human genome loci containing tandem repeats with varying copy numbers (loci ApoB, DX, S52, VWF, and YNZ22) were studied by means of polymerase chain reaction. The sensitivity determined by amplification of DNA mixture in dilutions was 1-2%. Based on the data obtained, various types of hemopoiesis recovery after bone marrow transplantation were determined; in most cases, a complete donor chimerism was revealed; in some patients, mixed chimerism; and in one case, a host type of hemopoiesis was found. An association between hemopoiesis type and further development of the disease was observed. PMID- 9206671 TI - [Comparison of localizations of insertions of phage-transposons D3112 and B3 into the Pseudomonas aeruginosa chromosome]. AB - Preferential chromosomal integration sites (sites of conservative transposition) of transposable phage D3112 were identified by means of pulsed-field gel electrophoresis and separation of restriction fragments of the Pseudomonas aeruginosa chromosome. Phase B3 was shown to differ from D3112 with respect to this trait. High site-specificity and regional specificity, typical for conservative transposition of D3112, can partially explain the absence of mutations in biosynthetic P. aeruginosa genes upon cell lysogenization with D3112 phage (unlike B3 phage). PMID- 9206672 TI - [Formation of sister chromatid exchanges and reparative DNA synthesis in workers exposed to nickel compounds]. AB - In workers from a nickel processing plant, residents of the surrounding industrial zone, and a control group, levels of sister chromatid exchanges (SCE, 33 individuals) and DNA repair synthesis (DRS, 79 individuals) were estimated. Individual variations in SCE level did not correlate to the duration of exposure to nickel compounds or the level of pollution. A statistically significant increase of SCE level among smokers compared to nonsmokers was revealed. Workers exposed to nickel compounds were demonstrated to have a statistically significant increase in the inhibition of DNA repair synthesis. PMID- 9206673 TI - [Cultivated keratinocytes on micro-carriers: in vitro studies of a new carrier system]. AB - Epidermal grafts from confluently cultivated keratinocytes have been used since the early eighties for the treatment of severe burns, where the shortage of donor sites for split-thickness skin grafts did not allow for adequate wound coverage. The difficult handling of these grafts as well as the advanced differentiation of their epithelial cells into a multilayer sheet poses a problem for their clinical application. The aim of the study was to characterize cultivated keratinocytes, as well as to observe their migration and proliferation from the MC onto a surface. Keratinocytes were isolated from human foreskin and cultivated in serum free and serum-containing medium according to a modified method by Rheinwald and Green. Collagen-coated Dextran beads were used as MC. The MC were colonized with keratinocytes using the Spinner culture technique. After seeding the colonized MC into culture flasks, their migration and proliferation was monitored regularly through immunohistochemical studies and measurement of the metabolic cell activity. Immunohistological staining proved that the cells isolated from human foreskin represent keratinocytes of the basal type. Keratinocytes, cultivated with serum-containing and serum free medium, both adhered to the surface of the MC, then migrated onto the surface of the flasks and proliferated to form a multilayer of epithelial cells. In the long-term, a flexible epithelial graft consisting of poorly differentiated keratinocytes should be available, which is simple to produce and easy to handle. This would be an alternative method for treating wounds, where the conventional multilayer epithelial graft (ET) is insufficient. PMID- 9206674 TI - [Comment on the contribution by L. Belusa, A.-M. Selzer and B.-D. Partecke: Description of Dupuytren disease by the Basel physician and anatomist Felix Plater in 1614]. PMID- 9206675 TI - [Surgical treatment of therapy refractory Sudeck's dystrophy by transaxillary decompression of the neurovascular bundle and sympathectomy. On the pathogenesis of Sudeck's disease]. AB - Since 1984, altogether nine cases of otherwise untreatable reflex sympathetic dystrophy (RSD) of the upper extremity were treated by transaxillary decompression of the neurovascular bundle with resection of the upper thoracic ganglia. This resulted in an immediate improvement of the local findings and the entire post-operative course of treatment as well as significantly improved functional results (seven excellent, one good, one fair). Average follow-up was 7.5 years. All patients returned to their original work. Clinical as well as intraoperative studies show that the acute edema of RSD is caused by a stenosis of the subclavian vein, which was proven in all cases by preoperative phlebography. Surgical decompression of the subclavian vein thus can result in an essential improvement of venous backflow and also significant improvement of the lesion-caused misproportion between increased arterial inflow and venous outflow with all resulting consequences for peripheral edema, subfascial pressure, microcirculation, perfusion of tissue and metabolism. By decompression of the subclavian artery and the lower plexus roots as well as by simultaneous transaxillary sympathectomy, the sympathetic efferents are drastically reduced or rather interrupted, leading to immediate improvement of the acute pain-syndrome. The so-called "individual predisposition" for RSD can mainly be attributed to a venous stenosis in the area of the subclavian vein and to an increase in sympathetic tonus by irritation of the lower parts of the brachial plexus and the postganglionic fibres accompanying the subclavian artery. The consequences for the pathogenesis of RSD are discussed in detail. PMID- 9206676 TI - [Effect of ACTH on long-term outcome of muscle regeneration after nerve injuries]. AB - An experimental investigation was performed to examine the influence of ACTH on the long-term results of muscle regeneration after nerve repair. Twenty albino rabbits ("White Russians") served as the animal model. The epi-perineural neurorrhaphy of the fibular nerve was carried out under microscopic control with 11x0 sutures. A computer-supported light-microscopic analysis of representative muscle-fiber diameters was performed for detection of nerve regeneration on the effector muscle of the peroneal nerve, the anterior tibial muscle, after 3, 4, 5 1/2, 7, and 8 months. In addition, the nerves were examined by light microscopy using not only standard techniques but also surgical immunohistological staining techniques (APAAP). The acceleration of nerve regeneration already described in the literature was confirmed by our investigations. However, the results observed after eight months were similar, regardless of the ACTH dosage applied. Muscle regeneration was not influenced by the injection of ACTH. It therefore does not appear justifiable to clinically apply ACTH for improvement of nerve and muscle regeneration. PMID- 9206677 TI - [A rare insertion variant of the extensor pollicis brevis tendon. A case report]. AB - Two brothers are presented with uni- and bilateral congenital palmar subluxation of the metacarpophalangeal joints of the thumbs as well as extensor lag. A hypoplastic extensor pollicis brevis tendon inserting atypically to the tendon of the extensor pollicis longus at the metacarpophalangeal joint level was found. In each case, arthrodesis of the metacarpophalangeal joint of the thumb was performed. This very rare variation of the insertion of the extensor pollicis brevis tendon is presented, surgical therapy and the pertinent literature reviewed. PMID- 9206678 TI - [Isolated traumatic dislocation of the thumb saddle joint]. AB - Isolated dislocations of the basal joint of the thumb are rare injuries. In case of early treatment and joint stability after reduction, immobilization in a scaphoid cast for six weeks is sufficient. Only in cases of delayed treatment or instability after reduction, stabilization by Kirschner-wires inserted percutaneously should be performed. PMID- 9206679 TI - [Results of surgical management of lunate malacia. A retrospective clinical study]. AB - We investigated the results of 53 patients in a retrospective study after surgical treatment for Kienbock's disease. Surgery included shortening of the radius (35 cases), cancellous bone graft (four cases), pisiform transposition (two cases), Swanson prostheses (two cases), tendon interposition arthroplasty (one case), STT-arthrodesis (six cases), denervation of the wrist (one case), and wrist arthrodesis (two cases). The average follow-up was 43 months (minimum 12, maximum 84 months). Criteria for rating the results were subjective satisfaction and pain profile. Objective criteria were X-ray findings and range of motion of the hand and wrist. We paid particular attention to the economic consequences for the patients. Based on our own results and the results in the literature, we recommend bone-grafting in the early Decoulx stages I and II. In stages II and III, we recommend in cases of ulna-minus variance the shortening of the radius in combination with a bone grafting; in case of an ulna-zero variance, we have the possibility of a radius wedge osteotomy or a pisiform transposition. Stage III, especially IIIb according to Lichtman, is suitable for STT-arthrodesis. In stage IV, we recommend total wrist arthrodesis, the denervation of the wrist can be combined with all the other methods or as a salvage procedure prior to wrist arthrodesis. PMID- 9206680 TI - [Surgical therapy of pronounced gout tophi in both hands. Case report]. AB - We report an 80-year-old male patient who had been suffering from gouty arthritis for the last seven years, both hands being deformed by multiple tophaceous deposits within soft tissue and bone. Immediate surgical therapy became inevitable when a ruptured tophus led to a severe infection of the right index finger. We describe the surgical procedure as well as the pathological findings and the clinical course. PMID- 9206681 TI - Comparative investigation of a streptovaricin-producing strain of Streptomyces spectabilis and its selectant. AB - Some differences were found in the ultrastructural, cultural and physiological biochemical properties between the parent strain Streptomyces spectabilis 1000 and its natural selectant S. spectabilis 1011-10, producers of streptovaricin. PMID- 9206682 TI - [Hepatitis B vaccination. Neglected vaccination is not ethically defensible anymore!]. PMID- 9206683 TI - [Hepatitis B vaccination of children and adolescents]. AB - In the 14 years that have passed since its introduction, the hepatitis B vaccine has proved to be highly immunogenic, effective and very well tolerated. The vaccination, previously performed only in persons at high risk of contracting hepatitis B, in particular medical personnel, has appreciably reduced the incidence of hepatitis B in this group. The incidence of the disease in the general population, however, has been reduced only marginally. Thus, the sole possibility of resolving the hepatitis B problem is to introduce general vaccination. To this end, the The Standige Impfkommission (STIKO) has included hepatitis B vaccination in the catalog of vaccinations for children and adolescents. The greatest importance must probably attach to vaccination of young children, in whom hepatitis B vaccination is carried out along the same lines as the diphtheria, pertussis and tetanus vaccination, within the framework of which it can therefore be administered, thus making possible a wide application. However, the 12-15 year-old age group should also be involved in the vaccination program, for this is the group on the threshold of the periods of greatest danger of infection, which begins with the initiation of first sexual relationships. PMID- 9206685 TI - [Frequently outside risk groups. Hepatitis B is still underestimated]. PMID- 9206686 TI - [Confusing games of public insurance with hepatitis B vaccinations. Interview by Dr. rer. nat. Anita Schweiger]. PMID- 9206684 TI - [Preventive vaccination of adolescents: is consent by parents always necessary?]. PMID- 9206687 TI - [Diagnostic strategies in rheumatology. 3: General laboratory diagnosis- detection of pathogenic infections]. PMID- 9206688 TI - [Hypertension and gastrointestinal diseases]. AB - Essential hypertension and gastrointestinal disease are two of the most common diagnoses made in the doctor's office. Since they may occur simultaneously, the differential therapeutic aspects of the respective therapies are of importance for every general practitioner. However, complex interactions between hypertension and gastrointestinal diseases often make the choice of the most suitable drug difficult. The present article provides an overview of the most common therapeutic regimens, identifies possible interactions, and should help the doctor to select the optimal treatment for the individual patient. PMID- 9206689 TI - [Mobile clinic for pygmies]. PMID- 9206691 TI - [Early intervention saves brain tissue. With a platinum spiral against aneurysms]. PMID- 9206690 TI - [Analog insulin requirements of the average patient. Interview by Dr. rer. nat. Anita Schweiger]. PMID- 9206692 TI - [Drug fraud--an unscrupulous business]. PMID- 9206693 TI - [Testicular torsion requires rapid diagnosis]. PMID- 9206694 TI - [Interferon-alpha in chronic myeloid leukemia]. AB - At the present time, the combination of interferon alpha and hydroxyurea represents the standard treatment of the chronic phase of CML. Using this approach, survival can be prolonged to a median of about five years. A precondition, however, is that treatment with interferon alpha is initiated as early as possible. The sole curative treatment continues to be allogeneic bone marrow transplantation. If no donor is available, autotransplantation with subsequent maintenance treatment with interferon alpha is an alternative approach. In view of the complexity of the treatment of CML, it is recommended that patients be referred to a center at the earliest possible time following diagnosis. PMID- 9206695 TI - [High dose chemotherapy with autologous stem cell transplantation]. AB - Subsequent stem cell transplantation makes it possible to increase the dose of oncological chemotherapy by a factor of between three- and ten-fold. For primary treatment, the superiority of this approach vis-a-vis standard doses seems to be highly likely in the case of recurrent highly malignant lymphomas, metastatic mamma carcinomas and plasmacytomas. To date, however, the most favorable time point for high-dose chemotherapy has not yet been definitively established-not has the patient group most likely to benefit or the most effective combination of cytostatics to be used. For this reason, such treatment, even of these diseases, should not be applied outside of controlled studies, and must be viewed even more critically in the case of other diseases for which data from randomized studies are not yet available. Despite a relatively low overall level of toxicity, peripheral blood stem cell transplantation (PBSCT) still has a mortality rate- even in large centers--of 2 to 5%. Any such treatment should therefore be given only in controlled studies and in suitable centers. PMID- 9206697 TI - [Diagnostic strategies in rheumatology. 2: Clinical diagnosis--stage 3]. PMID- 9206696 TI - [A goal is the dialogue between the established physician and the clinic. Interview by Dr. rer. nat. Anita Schweiger]. PMID- 9206698 TI - [Why does ACE inhibitor also reduce risk of myocardial infarct? Increased expression of endothelial nitric oxide synthase in atrial myocardium]. PMID- 9206699 TI - [Practicability of vaccinations in general practice]. PMID- 9206700 TI - [High dose ibuprofen in low back pain]. PMID- 9206701 TI - [General practice 2000--what should be done today?]. PMID- 9206702 TI - [The physician as alternative practitioner?--regrettably impossible]. PMID- 9206703 TI - [Acute thoracic aortic dissection with occlusion of the left coronary artery]. AB - Aortic dissection is the most common fatal condition that involves the aorta. Occasionally, symptoms mimic acute myocardial infarction leading to thrombolytic treatment. Accurate diagnosis in patients with chest pain is therefore essential. We describe a case of acute aortic dissection which resulted in myocardial infarction due to obstruction of the left coronary ostium. A 65-year-old female patient with no previous cardiac history was admitted to a local hospital because of severe chest pain of acute onset. Physical examination was normal except for a low blood pressure (90/50 mm Hg), heart rate 45 beats/min and parasthesia in both hands. The ECG showed sinus bradycardia with negative T-wave in VI and with 1 mm ST-segment elevation in V3. A chest X-ray was normal. Five hours later, the patient experienced once more severe chest pain followed by non-sustained polymorphic ventricular tachycardia (Figure 1). Another ECG showed bifascicular bundle branch block (right bundle branch block and left anterior fascicular block). The ECG was interpreted as showing acute myocardial infarction and treatment with intravenous streptokinase started. Since the patient remained severely hypotensive despite infusion of dobutamine, she was intubated, ventilated and transferred to our hospital. Cardiac catheterization showed acute dissection of the ascending aorta with an aortic intimal flap and an occlusion of the left coronary artery (Figures 2a and b). During catheterization, she suffered a cardiac arrest from which she could not be resuscitated. A postmortem examination confirmed the acute aortic dissection which reached to the ostium of the left coronary artery (Figures 3a and b, 4a and b) and an anterior myocardial infarction probably due to intermitted diastolic obstruction of the ostium of the left coronary artery by an aortic intimal flap. PMID- 9206704 TI - [Direct dilatation and emergency bypass operation of main branch occlusion in acute anterior wall infarct and cardiogenic shock]. AB - Occlusion of the left main coronary artery (LMCA) is the cause of myocardial infarction in about 0.04%. Those patients who do not die during the acute phase often do have a dominant right coronary artery with extensive collaterals to the left coronary artery. Because this is a very rare situation there are only some cases reports dealing with the management of these patients. A 60 years old woman was admitted to our hospital with the signs of an acute Q-wave anterior myocardial infarction. Within a few minutes after the arrival she developed a cardiogenic shock. Coronary angiography was performed immediately. The left main coronary artery was occluded and a big right coronary artery showed a significant stenosis. There were many collaterals from the right coronary artery supplying the left coronary artery. After information of the cardiac surgeons, primary angioplasty of the LMCA was performed in order to achieve hemodynamic stabilisation and to relieve symptoms. Reperfusion of the left anterior descendent coronary artery (LAD) could be achieved within 30 minutes. This led to hemodynamic stabilisation of the patient. But a significant residual stenosis of the LMCA remained and the circumflex artery was still occluded. In the meanwhile cardiac surgery was able to be performed and so the patient was transferred to surgery without further dilatation or stent implantation. Four venous grafts (LAD, first diagonal branch, circumflex artery and right coronary artery) were inserted. After 4 weeks the patient was in a good shape and could be discharged at home. Primary angioplasty seems to be an effective treatment in patients with acute myocardial infarction and an occlusion of the LMCA. But coronary bypass surgery is nearly almost necessary during the following period in order to achieve complete revascularisation and to improve survival. PMID- 9206705 TI - [Transplantation of aortic and mitral valves: permanent valve replacement without anticoagulation?]. PMID- 9206706 TI - [Quantitative determination of left ventricular myocardial perfusion with electron beam computerized tomography]. AB - Myocardial perfusion is one of the most important functional parameters of the heart. Presently various indirect methods are used to determine coronary blood flow or myocardial perfusion as inertgas-, thermodilution-, Doppler catheter- and radiopharmacological techniques. Electron-beam-computed-tomographical technology is able to perform CT data acquisition with a very short exposure time of 50 ms. Using this method it is not only possible to determine left ventricular volumes but also to measure myocardial perfusion in ml/100 g/min. The measurement of the left myocardial perfusion is performed using the short axis view. This position is obtained by moving the table 25 degrees to the patient's right and 15 degrees caudally. To determine the position of the left ventricle, a localization scan is obtained in multi-slice-mode using all for target-rings, thus obtaining 8 tomographic levels over 68 mm (each tomographic level having a slice thickness of 7 mm, with an interslice gap of 4 mm between each two adjacent tomographic levels). In this short axis position, using the multi slice flow mode with 3 target-rings and after administration of 50 ml of contrast medium intravenously with a flow of 3 ml/s, 6 tomographic levels are imaged. Each tomographic level is obtained 13 times at 80% of the R-R-interval at each 2 or 3 heart beat (ECG gated). The left ventricular myocardial contrast enhancement is measured by drawing manually the outline of the left ventricular myocardium using time density-software of the Imatron workstation. For calculation of the myocardial perfusion the so-called "slope method" is used and the results are expressed as the maximum slope of enhancement of the myocardium divided by the difference of the precontrast and peak CT-value in the left ventricle. The global myocardial perfusion is calculated as a mean of all evaluated tomographic levels. In this study left ventricular volumes as enddiastolic volume endsystolic volume and stroke volume were measured and ejection fraction and cardiac output calculated. The measurements were performed in the log axis view. This view is obtained by moving the table 15 degrees to the patients left in a horizontal position. In this long axis position 6 tomographic levels are imaged using the multi-slice cine-mode with 3 target-rings after administration of 50 ml of contrast medium intravenously with a flow of 3 ml/s. Each tomographic level is obtained 13 times starting at 0% of the R-R-interval (ECG-triggering). The exposure time is 50 ms with an interscan time delay of 8 ms. In 9 studied patients of whom one had 3 significant coronary artery stenotic lesions (> 50%), 2 patients had each 2 non significant stenotic lesions (< 50%) and 6 revealed nearly normal coronary angiograms. The mean global myocardial perfusion was 70 ml/100 g/min (min.32 and max. 116 ml/100 g/min). This mean value of 70 ml/100 g/min is reflecting 5% of the cardiac output supposing that the mean heart weight of these patients was 300 g. In this study the mean of the left ventricular muscle mass determined by the use of EBCT was 130 g. A comparative evaluation of coronary angiographic findings in these patients with the measured myocardial perfusion values revealed, that is not sufficient to look only at the absolute values of the measured myocardial perfusion. Furthermore it seems to be necessary to interpret these perfusion values with respect to the calculated cardiac output. Additional studies of well defined patients groups are necessary to determine normal values of myocardial perfusion at rest in patients with and without coronary artery disease. This seems to be important as comparative analysis of myocardial scintigraphic and EBCT-studies is difficult because of methodical inherent differences. The results of this study suggest that despite the presence of some beam hardening artifacts it is possible to measure myocardial perfusion using EBCT in patients with suspected coronary artery disease in the PMID- 9206708 TI - [Th1 cells, Th2 cells and atopic dermatitis]. AB - The immunological hallmark of atopic dermatitis (AD) is a Th1/Th2 dysbalance. The reaction to high molecular weight environmental allergens (e.g. pollen, house dust mites), production of IgE and activation of eosinophil granulocytes result from Th2 dominance. The Th2-cytokine interleukin-4 (IL-4) is necessary for IgE synthesis. Additionally, IL-4 inhibits the generation of Th1-cells. The marker cytokine of Th1-cells, interferon gamma (IFN gamma), exhibits reciprocal effects. It inhibits IgE synthesis and Th2 expansion, but supports Th1-cell growth. Beside the well known mechanisms of IgE-mediated immediate type reactions, the relevance of IgE for the pathogenesis of AD seems to be likely since the discovery of IgE receptors upon Langerhans cell surfaces. Langerhans cell-bound IgE may be possibly necessary for the presentation of high molecular weight aero-allergens. Analyses of Th subsets at different intervals after allergen challenge showed, that Th2-cells play an important role in the initial phase of inflammatory reactions whereas in later stages Th1-cells can be detected in greater numbers. PMID- 9206707 TI - [New developments in parameter-oriented roentgen densitometry perfusion analysis within the scope of heart catheter studies]. AB - X-ray densitometric evaluation of digital subtraction coronary arteriograms allows a qualitative and quantitative detection of contrast medium propagation through the epicardial coronary arteries, the capillary system and the coronary venous system. So-called "time-density-curves" (TDCs) can be generated following Lambert-Beer's law similar to indicator dilution curves by using contrast medium as the indicator. Several time and density parameters can be derived from these TDCs, which are related to local myocardial perfusion. Different animal validation studies have shown the applicability of this concept for in-vivo evaluation of coronary blood flow and myocardial perfusion. Nevertheless, absolute measurement of volumetric coronary blood flow or myocardial perfusion failed. Therefore, relative changes in coronary blood flow or myocardial perfusion in response to pharmacologically induced maximum hyperemia were measured and coronary or myocardial perfusion reserve was calculated as the ratio of hyperemic flow or perfusion divided by baseline values. Despite theoretical attractions for an application during routine cardiac catheterization, this densitometric approach did not get a wide acceptance. Primary reason for this limited use in specialized centers was the time consuming process of densitometric evaluation of the subtraction coronary arteriograms, which require digital cine angiography and necessitates enormous computer hard ware. This main limitation has been overcome since more powerful computer hard ware (processor speed, hard disk space, digitization boards) has become rapidly available during the last years at more moderate pricing and digital techniques today are state of the art in cardiac catheterization laboratories. In addition, soft ware program packages allowed an automatization of the digitization and densitometric evaluation process. These programs include ECG triggered cine image digitization with improved temporal resolution, semiautomatic definition of regions-of interest including definition of reference regions-of-interest for the detection of background density changes and quality-controlled densitometric parameter analysis. This progress made an application during routine cardiac catheterization feasible. In animal validation studies this improved X-ray densitometric approach for evaluation of local myocardial perfusion was validated versus colour-coded microsphere techniques. The time parameter "rise time", defined as the time from the start of local contrast medium induced density change to its maximum revealed a close correlation (r2 = 0.965) to the results of the microsphere technique over a wide range of perfusion. We have applied this technique before and after coronary interventions such as balloon angioplasty and stenting. Results documented an improvement of poststenotic myocardial perfusion reserve immediately after coronary balloon angioplasty and an additional improvement after adjunct coronary stenting. Only after stenting but usually not after coronary balloon angioplasty alone poststenotic myocardial perfusion reserve gained the intraindividual reference level, measured in a perfusion bed supplied by an epicardial coronary artery without stenoses. These results documented the functional benefit of coronary stenting on poststenotic myocardial perfusion in addition to the well known morphologic benefit with the creation of a larger and more circular conduit. PMID- 9206709 TI - [Brazilian pemphigus foliaceus (fogo selvagem)]. AB - Most of the clinical, histological and immunohistological features of fogo selvagem resemble those of idiopathic pemphigus foliaceus (PF). Both diseases are clinically characterized by small flaccid bullae evolving into to scaly and crusted lesions, sometimes with pustules, mainly in seborrheic areas of the skin. Mucosal surfaces are mostly spared. The main histologic feature of endemic pemphigus foliaceus is a subcorneal acantholytic blister. Standard immunofluorescence studies demonstrate intercellular IgG deposits throughout the entire epidermis. These IgG antibodies are mainly of the IgG4-subclass. Almost all patients have circulating IgG-autoantibodies in their serum directed against stratified epithelial desmosomes. The fogo selvagem autoantibodies and the PF antibodies are directed against the 160 kD desmosomal glycoprotein desmoglein 1 which together with plakoglobin (85 kD) forms a complex of adhesion proteins with desmosomes of stratified epithelia. Fogo selvagem occurs in endemic foci in some areas of Brazil and possibly in neighbouring South American countries, very often in children, adolescents and young adults. The etiology of fogo selvagem is still unknown. The frequent association with insect bites has lead to the concept of fogo selvagem being a transmissible disease with acquired immunity in adulthood. However, the infectious agent and possible vectors have not yet been identified. PMID- 9206710 TI - [The difficulty of ultrasound diagnosis of lymph node metastases of malignant melanoma in protracted tumor growth]. AB - Lymph node ultrasound examinations are performed during the follow-up of cutaneous melanoma in order to early recognise regional metastases. The excision of regional metastases is the only way to inhibit disseminated metastasis in a certain percentage of cases. Lymph node ultrasound has a sensitivity of 95% in the diagnosis of pathological lymph node changes. Sonographic findings normally show typical features of malignancy; only in some doubtful cases are further control examinations warranted over a period of 4-6 weeks. In violation of this rule, two cases are presented with a delayed onset of metastases causing difficulties in the diagnosis of malignant transformation. Four years after the excision of a nodular melanoma with 0.8 mm tumor thickness at the right lower leg, a suspicious lymph node change in the right groin was sonographically detected which did not fulfill all criteria of malignancy. Control examinations over a period of 1 year found only a further growth of 3 mm. Excision after 1 1/2 years showed a lymph node metastasis. The second case presented 9 years after the excision of a superficial spreading melanoma with 0.76 mm tumor thickness with a suspicious lymph node in the right axilla which similarly grew very slowly. The subsequent excision showed likewise a melanoma metastasis. A protracted growth may occur in thin malignant melanomas with late development of ultrasound features of malignancy. In doubtful cases an early biopsy is recommended. PMID- 9206711 TI - [Longitudinal study of the excretion of 1-hydroxypyrene in urine after external treatment with coal tar]. AB - In order to study the dermal uptake, time course, and urinary excretion of polycyclic aromatic hydrocarbons, the concentration of 1-hydroxypyrene in urine was determined by means of high performance liquid chromatography with fluorescence detection before, during, and after the topical treatment with coal tar in 19 patients suffering from prurigo simplex subacuta, microbial eczema, atopic dermatitis, eczematization after scabies, exanthematous lichen ruber, pityriasis lichenoides and cutaneous sarcoidosis. Beginning with a value of 6.04 +/- 2.06 micrograms 1-hydroxypyrene/g creatinine before treatment, the urinary excretion significantly increased during the therapy with coal tar (p < 0.0001 at 3rd, 5th, and 6th day of therapy). A maximum was reached at day 8 of topical treatment with a value of 584.35 +/- 191.96 micrograms 1-hydroxypyrene/g creatinine (p < 0.002). Already during treatment at day 10 there was a beginning decrease of 1-hydroxypyrene to 361.63 +/- 170.13 micrograms/g creatinine. After the end of treatment, the excretion further decreased reaching a value of 5.31 +/ 2.85 micrograms 1-hydroxypyrene/g creatinine at the 10th day after therapy. Skin carcinomas due to therapeutical use of coal tar occur extremely rarely and only after vergoten, non-controlled use. We suggest that the duration of exposure is the most important factor for the carcinogenic effect of coal tar. PMID- 9206712 TI - [Dapsone in granulomatous rosacea]. AB - We report on two patients with granulomatous rosacea and another patient with granulomatous perioral dermatitis who responded well to dapsone. Dapsone has a pharmacological double function as both an antibiotic and an antiphlogistic drug. Before the introduction of isotretinoin, dapsone had its place in the treatment of severe acne. To date, its use in granulomatous rosacea has not been described. When hematologic parameters are monitored, dapsone is considered a safe and cost effective drug, especially in countries where isotretinoin is not readily available. However, the definite value of dapsone in granulomatous rosacea should be established by a controlled study. PMID- 9206713 TI - [Reversible increase in photosensitivity to UV-B caused by St. John's wort extract]. AB - A 61-year-old woman with depression developed recurring elevated itching erythematous lesions in light-exposed areas after taking St. John's Wort-extract for three years. Routine patchtesting did not reveal any relevant reactions and photopatch testing was negative. Using a systemic oral photoprovocation test with St. John's Wort, we were able to demonstrate a decrease of the MED-UVB which was reversible after withdrawal of the medication. PMID- 9206714 TI - [Juvenile hyaline fibromatosis]. AB - Juvenile hyaline fibromatosis is a rare autosomal recessive connective tissue disease first described in 1873 by Murray. Major diagnostic criteria are multiple cutaneous tumors and gingival hypertrophy; minor criteria include contractures, osteolytic lesions and a positive family history. After a normal perinatal period at the age of 6 months our 24 year old patient developed gingival hypertrophy. During the first months of life several skin coloured nodules had been noticed on the neck and in the perianal area. At the age of 15 months, these nodules began to appear more rapidly, both spontaneously and posttraumatically. The patient showed normal development, but the lesions progressed. By the age of 15 years, the patient had extensive deformities and was unable to walk and move by himself. Both his sisters and the unrelated parents had no lesions. Essential for the diagnosis are the clinical picture and the histology. Electron microscopy is helpful to support the diagnosis. Defective connective tissue proteins such as chondroitin, collagen and mucopolysaccharides are probably the pathological defect. A therapy is so far unknown. PMID- 9206715 TI - [Juvenile Adamantiades-Behcet disease in decreased stimulation with anti-CD3 monoclonal antibody]. AB - A 12 year old boy developed the complete symptom complex of Adamantiades-Behcet's disease over a two-year period. He presented with the mucocutaneous variant with recurrent to persistent oral ulcers which extended into the pharynx, recurrent genital ulcers, perianal lesions, and a positive pathergy test. Ocular involvement and other symptoms associated with the disease were absent. However, the early onset of the disease and the male gender indicated a bad prognosis. The patients father deficiency of suffered from bronchial asthma with recurrent respiratory infections. An IgG-3 subclass was detected. Lymphocyte transformation tests showed markedly diminished response to stimulation with anti-CD3 monoclonal antibody in both patients, while response to PWM, CoA and PHA was normal. In addition, the concentration of serum soluble interleukin 6 receptor was reduced in both patients. PMID- 9206716 TI - [Kaposi sarcoma in cyclosporin A therapy of actinic reticuloid]. AB - A 76 year old patient with actinic reticuloid was treated with Cyclosporin A. 11 months after beginning the immunosuppressive therapy, he developed lymphoedema and livid nodular skin lesions of the right leg, which histologically showed Kaposi's sarcoma. After stopping the Cyclosporin A and intralesional therapy with Interferon alpha, no regression of the Kaposi sarcoma could be seen. The characteristics of Kaposi sarcoma arising during immunosuppressive therapy and the differences in incidence, risk groups and distribution pattern compared to the classic sporadic and the AIDS-related Kaposi sarcoma will be described. The development of Kaposi sarcoma during immunosuppressive therapy for actinic reticuloid has not been previously described. PMID- 9206718 TI - [Multiple eccrine hidrocystomas in Parkinson disease]. AB - We describe clinically and histologically typical eccrine hidrocystomas on the eyelids and forehead of a 60-year-old woman which were successfully treated with topical atropine. Concomitantly the patient had long-standing Parkinson's disease with typical facial seborrhea and hyperhidrosis. In this report, we review the literature about eccrine hidrocystomas and discuss the question of a causal relationship with Parkinson's disease in our patient. PMID- 9206717 TI - [Tufted hair folliculitis]. AB - A case of tufted hair folliculutis presenting as circumscribed, tender and inflamed areas in the occiput with residual tufted follicles in a 28-year old man is reported. Tufted hair folliculitis is a characteristic localized scarring bacterial folliculitis of the scalp due to Staphylococcus aureus. Histopathological studies reveal perifollicular inflammation around the upper portions of the follicles sparing the hair root level. Within areas of inflammation, several follicles converge toward a common follicular duct with a widely dilated opening. Currently, tufted hair folliculitis is considered a variant of folliculitis decalvans of Quinquaud. Staphylococcal infection is believed to be an initial causative factor, and underlying differences in follicular anatomy or host response may be important in determining which reaction pattern occurs in an affected individual. The development of atrophy with loss of adnexal structures (in folliculitis decalvans) or of hair tufts (in tufting folliculitis) may depend upon the depth and destructive potential of the inflammatory process. The therapeutic approach is problematic; prolonged treatment with oral antibiotics may stabilize the disease, but good and at times more definitive results (as in the presented case) have been reported after radical surgical excision of the involved areas. PMID- 9206719 TI - [Report on the 2nd Baltic-German Symposium of Andrology and Reproduction Medicine]. PMID- 9206720 TI - [Hans von Hebra (1847-1902) on his 150th birthday. An epitaph]. PMID- 9206721 TI - [Diabetic foot]. PMID- 9206722 TI - [T-helper cells: the key to the pathogenesis of psoriasis?]. PMID- 9206723 TI - Fairy-tales in psychotherapy. AB - Fairy-tales, like mythologies, can be found all over the world containing the same motif and chains of motifs. In this paper I have presented some theories on the occurrence of this archetypal phenomenon ranging from the old migration theory to Sheldrake's theory of morphogenetic fields. I have then tried to show how fairy-tale-motifs can appear in various ways in analytical therapy, often in hidden forms. We find them in patients' dreams as well as in their fantasies and associations. If the therapist is open to them they will also appear in his or her amplifications. He or she might then take note of the fairy-tale or point it out to the patient; in the latter case it might provide better access to the patient's problems and complexes as fairy-tales have an emotional completeness because of their pictorial character. Finally I have described the favourite fairy-tale of one of my patients and related it to his symptoms, his central complex and his personal ways of experiencing and behaving. This survey of how fairy-tales can be used in therapy with children and with adults far from exhaustive. PMID- 9206724 TI - Hyaluronan synthases. PMID- 9206725 TI - [Transplantation of retinal cells]. PMID- 9206726 TI - [Genetics of macular degeneration]. AB - Hereditary macular degeneration comprise a large group of disorders characterized by a preferential loss of central vision due to degenerative changes in the macular area of the retina. The primary causes underlying these degenerative processes are unknown. Recent approaches in molecular genetic research are most promising in providing new information for the understanding of the basic biochemical defects causing the specific macular phenotypes. Over the last few years an increasing number of hereditary degenerations has been mapped to specific chromosomal regions as a prerequisite for the final cloning of the respective disease gene. The knowledge of the function of the disease gene products will then permit further insight into the functional aspects of the normal and diseased retina. This will be most important for clinical diagnosis, management and treatment of patients with macular degenerations. PMID- 9206727 TI - [Retinitis pigmentosa--clinical, genetic and pathophysiologic aspects]. AB - Retinitis pigmentosa defines a genetically heterogenous group of disorders characterized by degenerations of photoreceptors and pigment epithelium. This article reviews our current knowledge of the genetical, clinical and pathophysiological aspects of this disease complex. Therapeutic concepts under current investigation are discussed as well. In recent years tremendous new insights have been made using molecular techniques for the investigation of retinal dystrophies. Ophthalmoscopically very similar patterns of photoreceptor dystrophies have been related to different gene mutations. In contrast, mutations in a single gene may cause different clinical patterns of photoreceptor dystrophies. Therefore, these recent results suggest that a reclassification of retinal dystrophies on the basis of their genetic origin may be favourable. In the future molecular genetics and the recent developments may play an increasing role for clinical classification and evaluation of photoreceptor dystrophies. The continued clinical and experimental research on hereditary disorders may help to elucidate further the wide disease spectrum and thereby developing new classifications and efficient therapeutic concepts. PMID- 9206728 TI - [No-stitch phacotrabeculectomy using modified punch technique]. AB - BACKGROUND: Combined cataract-glaucoma surgery is associated with a higher rate of intra- and postoperative undesired reactions than if each procedure is performed separately. We therefore investigated weather use of a special punch for trephination of a fistula opening in the area of the scleral tunnel could both reduce the rate of complications and improve regulations of pressure. METHOD: The modified procedure was performed in 22 patients. After phacoemulsification and implantation of a foldable lens through a 3.3 mm wide scleral tunnel, a 1.1 mm wide trephination opening was made in the lower wound flap using a specially designed punch. Without stitching the tunnel, the fistula was checked and the operation ended by suturing the limbus-based conjunctival flap. RESULTS: The rate of immediate postoperative complications was markedly reduced by that procedure. A postoperative hyphema did not occur in any patient. The incidence of fibrin exsudation was also considerably diminished. With regard to the medium-term stabilization of pressure, our method was slightly better than those reported in other studies. CONCLUSIONS: In light of the good results achieved with the modified procedure described here for the first time, a rethinking of the strict indications for this type of intervention in patients with cataract and simultaneous open-angle glaucoma is warranted. PMID- 9206729 TI - [Recent developments in lacrimal duct endoscopy]. AB - BACKGROUND: At the moment digital dacryocystography yields the most exact results in lacrimal diagnostics. The main disadvantage lies in the dependency of high tech X-ray systems and an angiography unit, which raise enormously the costs of the examination. Recently a new diagnostic system of the lacrimal pathway is available. PATIENTS AND METHODS: Miniaturizing of endoscopes and new developments in the fiberoptic technology made it possible to introduce these mini-endoscopes into the canaliculi and perform antegrade examination of canaliculi, lacrimal sac and ductus nasolacrimalis. 86 patients agreed to this examination. With this examination a direct visualisation of the mucous membrane of the lacrimal pathways and possible pathological alterations were possible. RESULTS: In case of axial illumination and syringing of the lacrimal system good visibility can be achieved and an exact examination is possible. Especially for endoscopy modified probes of Bangerter provide an exact examination at narrow areas and areas of bendings too. Different reasons for stenosis of the lacrimal pathways could be identified, especially 2 cases of a lacrimal sac tumor. CONCLUSIONS: The use of this new miniendoscope provides a very good examination under direct visualisation of the lacrimal pathway independent of a high tech X-ray equipment. The goal is to use this new endoscope on the one hand only for diagnostic reasons as endoscope and on the other hand in combination with a laser fiber to perform an antegrade laser dacryocystorhinostomy. PMID- 9206730 TI - [Echographic diagnosis of efferent lacrimal ducts with contrast media]. AB - BACKGROUND: Affections of the lacrimal drainage system are often seen by ophthalmologists. Inspection, palpation, diagnostic rinsing and sounding can distinguish anatomical stops before or after tear sac. For final diagnostics however more apparative examinations are necessary. The ultrasonic examination with contrast media is a simple method for diagnostics of affections of the lacrimal drainage system besides the dacryocystography, the scintigraphy and other ones. PATIENTS AND METHODS: On 12 patients with a dysfunctional lacrimal drainage system and 12 normal controls the ultrasonic examination with instillation of contrast media: silicon oil, glycerine, dispersions of almond oil and viscoelastic substances was performed. All examinations were performed with the 13 B-scan. RESULTS: The lacrimal drainage way was detectable from the canaliculus to the middle nasolacrimal duc CONCLUSION: An additional advantage of the ultrasonic examination with contrast-media is the neglect of radiation, the simple and often repeatable examination method, especially the enhancement of the contrast of different valves and stenoses and mucinous deposits in stationary anatomical variations and dynamic defects. PMID- 9206731 TI - [Keratoconjunctivitis sicca in patients with lipoprotein (a) elevation]. AB - BACKGROUND: Elevated Lp(a) levels are known as risk factor for premature coronary artery disease and apoplexy. To our knowledge ophthalmologic alterations have not been described in combination with elevated Lp(a) levels. We examined whether patients with LP (a) elevation had an increased incidence of keratoconjunctivitis sicca. PATIENTS AND METHODS: 27 patients, 16 women and 11 men were examined. The Lp(a) value was in all patients definitively pathological. The following examinations were performed: history, slitlamp examination, Schirmer-test, impressioncytology, fluorescein staining of the cornea, applanation tonometry and examination of the ocular fundus. RESULTS: The results show a pathologic Schirmer test in 70.3% of the eyes, in 81.5% a pathologic impressioncytology, and in 48.1% a pathologic fluorescein staining. The Schirmer-test is not sufficiently sensitive and specific in diagnosing the dry eye, whereas impression cytology is highly sensitive and specific. CONCLUSIONS: We could show, that in more than 80% of the examined patients at least one sign of keratoconjunctivitis sicca was found; however we did not find a correlation between the height of the Lp(a) level and the degree of pathologic impression cytology. Further examinations are needed to investigate the causal connection between keratoconjunctivitis sicca and Lp(a) elevation. PMID- 9206732 TI - [Visual acuity and use of eyeglasses after implantation of a diffractive multifocal lens]. AB - BACKGROUND: The implantation of a diffractive multifocal lens (dMIOL) as alternative to a monofocal lens is justified if after surgery there is practically no need to wear glasses. PATIENTS AND METHODS: 31 patients had an implantation of a total of 35 dMIOLs (3M 815 LE). We evaluated the visual acuity, the refractive data and the patients' attitude to wearing glasses. The average age was 67.0 +/- 11.8 years. Follow up took place after 18.7 +/- 5.4 months. RESULTS: The mean value of the uncorrected distance acuity was 0.59 +/- 0.17 and the corrected distance acuity 0.96 +/- 0.13. The uncorrected near acuity amounted to Jg 2.40 +/- 0.94, best distance correction was Jg 1.49 +/- 0.55. The patients still accepted an average of 0.68 +/- 0.37 dpt for the best near correction (near vision over the diffractive near focus) and thus achieved Jg 1.46 +/- 0.55. At the best distance correction plus 3.5 dpt, the near visual acuity was improved to 1.03 +/- 0.17. 54.8% of the patients indicated that they did not use glasses at all. 32.3% stated that they only used glasses for reading. 9.7% wore bifocals all the time, and 3.2% always used glasses for the distance. CONCLUSIONS: Regarding distance vision, the dMIOL is equivalent to monofocal lenses. Without any correction the results of the dMIOLs for the near vision are superior to monofocal lenses. Glasses can be dispensed with if the uncorrected visual acuity of the operated eye is at least 0.6 Jg 2-3 after surgery and the other eye too, does not need any correction. In case the postoperative visual acuity is worse, good visual acuity of the other eye may render glasses unnecessary. Part of the patients put up with a correctable loss of visual acuity in order not to become dependent on glasses. The need to wear glasses may be decreased considerably by implanting a dMIOL in both eyes and by avoiding postoperative refraction errors. PMID- 9206733 TI - [Tyndallometry and cell count in the anterior chamber in retinal detachment]. AB - BACKGROUND: Aim of this study was the investigation of changes of aqueous flare and cells in eyes with retinal detachment. PATIENTS AND METHODS: We examined 62 eyes of 61 patients with retinal detachment (56 eyes with rhegmatogenous retinal detachment, 6 eyes with traction retinal detachment; 51 eyes without and 11 eyes with clinical signs of proliferative vitreoretinopathy (PVR) stage C) with the laser flare-cell meter (LFCM) and compared the results with clinical findings and with the results of a control group. RESULTS: The flare values as well as the cell count of all eyes with retinal detachment were significantly elevated (p < 0.0001) in comparison to those of the control group. In eyes with PVR a significant elevation of flare values in comparison to eyes without PVR appeared (p = 0.0052). A slight correlation was found between the extension of the detachment and the flare values and the cell count, respectively. In cases with rhegmatogenous retinal detachment a significant elevation of the cell count was found if the largest break was localized in the upper hemisphere (p = 0.03) or if the macula was affected (p = 0.02). In rhegmatogenous retinal detachment, the cell count correlated slightly with the age of the patients (r = 0.3, p = 0.02), and flare values correlated with the height of the detachment (r = 0.28, p = 0.04). CONCLUSIONS: Our results demonstrate a breakdown of the blood-ocular barriers in eyes with retinal detachment and an elevation of corpuscular elements in aqueous as manifestations of pseudouveitis. In eyes with PVR the alteration of blood-ocular barriers seems to be more extensive than in eyes without PVR indicating a possible role of the LFCM for early detection of PVR. PMID- 9206734 TI - [Correlation of ultrasound biomicroscopy with histological findings in diagnosis of giant cell arteritis]. AB - BACKGROUND: A biopsy of the temporal arteries is still the appropriate method to prove the diagnosis of giant cell arteritis. We evaluated the potential use of high-resolution ultrasound-biomicroscopy in the diagnosis of giant cell arteritis. PATIENTS AND METHODS: In a prospective study we examined 16 patients (8 women and 8 men) with a mean age of 71 years with the clinical suspicion of a giant cell arteritis. Additionally to the clinical examination the temporal arteries were imaged in all patients using the ultrasound-biomicroscopy (Zeiss Humphrey Instruments). The results were correlated to the histopathologic changes of the temporal arteries excised bilaterally at the same location. RESULTS: Histopathological evaluation revealed a granulomatous arteritis in 4 out of 16 examined patients. The temporal arteries of these patients also showed characteristic changes using ultrasound biomicroscopy like middle-reflective shadowing of the arterial lumen and a condensation and enlargement of the muscularis media. Ultrasound-biomicroscopy allowed a precise evaluation of the temporal arteries due to a high-resolution sonographic image. The morphological differentiation between a normal and an affected artery was possible. A positive correlation between histopathological and clinical findings was seen in all patients. CONCLUSION: In this preliminary study the ultrasound-biomicroscopy seemed to be an appropriate non-invasive tool for the morphological imaging and evaluation of temporal arteries. PMID- 9206735 TI - [Effect of panretinal photocoagulation on the ocular pulse curve]. AB - BACKGROUND: Most studies on the effects of panretinal photocoagulation (PRPC) on ocular circulation are concerned with retinal circulation. In the present study, we examined the ocular pulse curve after PRPC and determined pulsatile ocular blood flow (an indicator of choroidal circulation) and ocular perfusion pressure. PATIENTS AND METHOD: In 10 patients with diabetes mellitus (8 with type II and 2 with type I; mean age 64 yrs) and severe, hitherto untreated, bilateral proliferative diabetic retinopathy, an intensive unilateral PRPC with 1500 argon laser burns (spot size: 500 mu) was performed in 2 sessions (interval: 3 wks). Before and-in 3-wk intervals-up to 9 wks after onset of treatment, an ocular pulse curve was recorded using oculo-oscillo-dynamography, and the following variables were determined: ocular pulsation amplitude (OPA), pulsatile ocular blood flow (POBF), systolic ophthalmic artery pressure (SOAP), and systolic ocular perfusion pressure (SOPP). RESULTS: PRPC led to a reduction of OPA and POBF in relation to the untreated contralateral eyes. 3 wks after the 1st coagulation, the reduction averaged 20% (OPA) and 19.1% (POBF). The maximum reduction was found 9 wks after onset of treatment (6 wks after the 2nd coagulation) and amounted to 29.9% and 29.2%, respectively. The differences between photocoagulated and untreated eyes were highly significant on average (P < 0.01; ANOVA) as well as for the time course (P < 0.001). SOAP and SOPP were not changed significantly during PRPC. CONCLUSIONS: OPA and POBF are determined by the cardiac cycle-related intraocular volume changes depending predominantly on choroidal blood flow. Thus, the reduction of POBF after PRPC is indicative of a lowered pulsatile choroidal blood flow. The morphological substrate is probably the choriocapillaries closure after photocoagulation described in the literature. PMID- 9206736 TI - [Fornicate structure of blind saccular lymph vessel segments in the conjunctiva- scanning electron microscopy studies]. AB - BACKGROUND: The accepted fundamental element of the initial lymphatic system is a tubular structure (lymphatic capillary), which is present either in the form of a finger-shaped blind-ending protuberance, or in the nature of a plexiform composite arrangement. We were able to demonstrate experimentally that the finger shaped protuberances, which are known as "initial segments" are in fact temporary filling states. As the filling process continues, these vascular elements become intermediate segments, via which other parts of the vascular network take up the dye. To date there have been no investigations of the internal structure of these apparently blind-ending vessel segments. In this study, lymphographically represented "terminal segments" should be cut away for the purpose of examining the fornix by scanning electron microscopy. Is the internal surface of the fornix regular and unbroken, or are there fissure-like structures which could explain the observed filling processes? MATERIAL AND METHODS: In conjunctiva of bovine eyes (n = 80), interstitial double-contrast lymphography (Berlinblue solution/air) under a slit-lamp microscope was used specifically to search for finger-shaped terminal segments. The conjunctivae had previously been prepared by fixing in polymeric resins. A proportion (n = 21) of the specimens were amenable to examination under the scanning electron microscope. RESULTS: Notwithstanding the observed great variation in the shape of the lymphographically obtained blind ending structures-in the form of terminations shaped variously like fingers, balloons, domes, pistons or pyramids, terminations with two humps, and terminations shaped like spear heads-scanning electron microscopy revealed within the fornices many relatively uniformly shaped structures in the form of fissures, configured with lips and saw-tooth edges, rather like zip fasteners. These findings are suggestive of preformed connections to neighbouring segments. This appears to be another element, in addition to the familiar flap-like structures, for controlling the circulation of lymph. Does the "initial part" of the lymphatic system now have to be redefined? PMID- 9206737 TI - [Cutaneous T-cell lymphoma of the mycosis fungoides type in the area of the eyelid and lacrimal ducts]. AB - PATIENT: A 62-year-old former miner with silicosis of the lungs but otherwise in good general condition presented with a solid nodule in the nasal left lid area for a duration of three months. Because of a central ulceration the reference diagnosis was basalioma. The tumour infiltrated the nasal part of the upper and lower eyelid and the tear ducts so that these were unrinseable. Similar lesions have been present since two years in other skin regions. METHODS: Two cutaneous biopsies confirmed the diagnosis of a Mycosis fungoides without detectable expression of the CD30-antigen. Medical investigation finally revealed hepatosplenomegaly and cervical, inguinal and abdominal lymph node involvement. A lymph node biopsy three months after presentation again showed a T-cell-lymphoma which was CD30-positive now. THERAPY: Systemic polychemotherapy was started. The lid lesions completely resolved, and the tear ducts were rinseable again. PMID- 9206738 TI - [Massive foreign body reaction after intraocular penetration of hot, liquid plastic]. AB - PATIENT: A 32-year-old male had an injury of his right eye by hot liquid plastic (Ultradur R). Intraoperatively, the eye was found broadly penetrated and filled up by more than 50% with ragged pieces of plastic. Computed tomography three days after wound closure revealed remaining foreign body material. As function was totally lost no further operation was considered. Seven weeks after the trauma, the eye was enucleated because of a painful phthisis. Histology showed numerous giant cells. CONCLUSIONS: Hot liquid plastic may perforate the eye and fill it up to a great extent. Due to intraocular cooling, pieces with sharp edges may develop, thus producing further (mechanical) tissue damage. Intraocular plastic is able to induce a massive foreign body reaction. It may be detected easily by computed tomography. PMID- 9206739 TI - [In sudden blindness: nitroglycerin? Reflections on therapy of acute embolism embolism of the central retinal artery]. PMID- 9206740 TI - [Hippel-Lindau syndrome. Clinical and genetic aspects of angiomatosis retinae]. PMID- 9206741 TI - [Contract physician services on private accounts and possibilities for bypassing the reimbursement conditions]. PMID- 9206742 TI - [Video stereophotography of the optic papilla. A practice-oriented documentation]. AB - Video documentation of the posterior pole using a camera installed in the ocular part of a slitlamp provides visualization of the optic disc from two different angles by changing the ocular part with the camera from the right to the left. Using the appropriate video printer program (split 4) two pictures of the right optic disc are stored on the upper half, one shooting from the right the other from the left. The lower half of the print is reserved for the left optic disc. Thus stereoscopic perception of the optic discs can be achieved. By this means a simple procedure has been provided for improved follow up of glaucoma patients. PMID- 9206743 TI - [Ocular pathology of child abuse]. AB - BACKGROUND: In spite of a growing awareness in the population most cases of child abuse remain probably undetected. Ocular changes in this syndrome are manifold. Sometimes ocular signs can help to substantiate the suspicion of child abuse. On the other hand the ophthalmologist may be the first physician to be contacted. Thus, he plays an important role in diagnosis. Though there are a couple of clinical descriptions morphological data are almost completely missing in the German literature. PATIENT: A two-year-old girl died two days after severe abuse because of widespread intracranial hemorrhages with brain stem insufficiency. At autopsy both eyes were enucleated and sent for histological investigation. RESULTS: The anterior segments were unremarkable. Multiple hemorrhages were found in the inner retina bilaterally. Moreover there were preretinal, intrachoroidal, intrascleral (area of the circle of Zinn-Haller) and subdural hemorrhages. One eye showed a circular, perimacular fold of the central retina and a hemorrhagic retinoschisis. CONCLUSIONS: Intraretinal hemorrhages alone are typical though not pathognomonic for the "battered-child syndrome". However, in combination with a crater-like appearance of the central retina, a hemorrhagic retinoschisis; and intrascleral hemorrhages in the area of the circle of Zinn-Haller they suggest child abuse almost with certainty. The pathogenic mechanisms leading to the observed fundus changes lack definite clarification. The date of violence which is essential for legal prosecution can be difficult to evaluate on morphological grounds alone. PMID- 9206744 TI - [Acute and chronic immune reactions after penetrating keratoplasty with normal immune risk]. AB - BACKGROUND: For diagnostic and therapeutic reasons it is important to differentiate between acute and chronic immune reactions. Up to now in the literature as well as in clinical follow-up such a differentiation has been performed only very insufficiently. We analysed retrospectively frequency and type of immune reactions after penetrating keratoplasties with normal immune risk in order to have a data basis for comparison with the corresponding data from our high risk keratoplasties. PATIENTS AND METHODS: The clinical courses of 646 penetrating keratoplasties with good prognosis performed between 11/1986 and 6/1994 were analysed. The mean patient age was 58 (12-89) years. Only endothelial and stromal immune reactions were recorded. RESULTS: 18% of the grafts suffered from at least one immune reaction during the first 3 postoperative years (Kaplan Meier value). 94% of the grafts without immune reactions remained clear in contrast to 45% of the grafts with immune reactions during this period of time (Kaplan Meier values, Log Rank Test: p < 0.001). 81 immune reactions were observed after 62 keratoplasties. 45 immune reactions (56%) had an acute course (43 endothelial, 2 stromal). 36 (44%) were chronic (31 endothelial, 5 stromal). 93.7% of the grafts without clinical signs of immune reactions, 100.0% of the grafts with only chronic immune reactions and 38.7% of the grafts with only acute immune reactions were clear 3 years postoperatively (Kaplan Meier values). No graft with a combination of acute and chronic immune reactions was clear 3 years postoperatively. 56% of all acute immune reactions occurred during the first postoperative year, 82% during the first 2 and 91% during the first 3 postoperative years. For chronic immune reactions the corresponding values reached 51%, 94% and 100%. CONCLUSIONS: After penetrating normal-risk keratoplasty acute immune reactions occur more often than chronic immune reactions, but the latter are in fact far more frequent than anticipated. If they are diagnosed in time and treated correctly they do not lead to graft failure in the mean run. Both types occur predominantly during the first 3 postoperative years and only rarely thereafter. PMID- 9206745 TI - [Modification of axial length and astigmatism by scleral buckling surgery]. AB - BACKGROUND: Scleral buckling surgery for retinal detachment alters the shape of the globe resulting in changes of the refractive state of the eye. MATERIALS AND METHODS: In a prospective study of 52 eyes with retinal detachment we examined changes of corneal astigmatism and axial length induced by encircling buckling or segmental buckling spanning two quadrants. We compared our results with those found in literature. RESULTS: In most patients we found a shortening of the globe, rather in cases with encircling buckles than in the cases with segmental buckles (parallelly to the limbus). We also saw changes in astigmatism postoperatively in all patients. There was no statistically significant difference between patients with encircling and segmental buckles fixed parallelly to the limbus, neither in the change of axial length nor in the change of astigmatism. CONCLUSIONS: In all cases of scleral buckling procedures changes of corneal astigmatism and axial length are to be expected. So our results differ from those of other authors who did not always find a change of astigmatism. We suppose that the authors of the publications which differ from our results did not consider the change of the corneal axis by using the vector method. PMID- 9206746 TI - [Cryocoagulation in therapy of proliferative diabetic retinopathy]. AB - BACKGROUND: The importance and indication of panretinal photocoagulation in proliferative diabetic retinopathy is well established. In contrast the indication of cryotherapy in this disease is more controversial especially in regard of new indications for early vitrectomy. The present study was performed to characterize the clinical possibilities and limitations of cryotherapy in complicated proliferative diabetic retinopathy. PATIENTS AND METHODS: In 231 patients with proliferative diabetic retinopathy and vitreous hemorrhage limiting further photocoagulation the visual outcome and diabetic retinal changes were observed before and after cyrotherapy (15-20 effects) of the ophthalmoscopically visible peripheral retina. RESULTS: After cryotherapy regression of active proliferations could be seen in 70% of the patients. Resorption of vitreous hemorrhages could be found in 80% of the patients. This was associated with improvement in visual acuity in 50-60% of the patients. Loss of vision was caused due to tractional detachment in 20% of the patients and due to further vitreous hemorrhages in 10% of the patients. Comparison of retinal changes between patients with worsened visual acuity and patients with increase in visual acuity demonstrated the preoperative fibrotic status of disc neovascularisation as the most important prognostic factor. The development of central tractional detachment was significantly higher in patients with preoperatively partly regressed disc neovascularisation. CONCLUSIONS: Cryotherapy of the peripheral retina in proliferative diabetic retinopathy with vitreous hemorrhages is therefore only indicated after ophthalmoscopical or echographical exclusion of peripapillary fibrosis and retinal traction and with sufficient visibility of the peripheral retina for the application. PMID- 9206747 TI - [Clinical aspects and histopathology of caruncular tumors]. AB - BACKGROUND: Histologically the lacrimal caruncle represents a transition zone between the conjunctiva and the skin containing cutaneous, conjunctival and lacrimal elements. This accounts for a variety of histopathological changes. METHODS: We conducted a retrospective study on the histopathological findings of 65 caruncular lesions which were referred for excisional biopsy between 1974 and 1995 to the Department of Ophthalmology at the University of Heidelberg. The age of the patients ranged from 10-87 years (mean 42 +/- 18.2). RESULTS: Histopathologic diagnoses included 29 (45%) nevi, 15 (23%) papillomas, 7 (11%) chronic inflammatory lesions and 4 (6%) epidermoid cysts. In addition, 4 (6%) semimalignant and malignant tumours were found including one basalioma, one squamous cell carcinoma and two Kaposi-sarcomas. Among the remaining 8% a dermoid cyst, an accessory lacrimal gland with a retention cyst, an epithelial implantation cyst, a teleangiectatic granuloma and an oncocytoma were diagnosed. CONCLUSION: These findings suggest that malignant caruncular lesions are rare, however excisional biopsy appears prudent in order to establish the histological diagnosis. PMID- 9206748 TI - [Photoscreening for early detection of amblyogenic eye changes]. AB - BACKGROUND: Congenital and early acquired ocular changes impairing the optic input induce amblyopia when left untreated. Amblyopia treatment must start early to be efficient. Therefore it seems necessary to employ screening tests in preverbal childhood. PATIENTS AND METHODS: The reliability of two commercially available photoscreening devices, the "Visiscreen 100" (Vision Research Corp.) and the "MTI-Photoscreener" (Medical Technology Inc.), was tested. 180 children from a kinder-garten and 120 infants from our outpatient clinic were screened. The results were compared to the findings of a full ophthalmologic and orthoptic examination. RESULTS: The efficacy of the photoscreening depended on the skill of the examiner and on the age of the children tested. The rate of interpretable photographs was 94% in the older group and 63.3% in the infants. The mean sensitivity for detection of amblyogenic factors was 63% in the older and 80% in the infant group. The mean negative predictive value was 90%, and 75%, respectively. CONCLUSION: Modern photoscreening techniques can help to detect amblyogenic factors in early childhood. However, in addition to the non interpretable photographs, about 20% of the affected children are missed. Therefore, photoscreening cannot be recommended for countries with a high number of ophthalmologists, such as Germany. Instead, an ophthalmologic and orthoptic investigation in early childhood would be preferable. PMID- 9206749 TI - [Local cyclosporin A therapy of nummuli after epidemic keratoconjunctivitis--case report]. AB - BACKGROUND: Steroid therapy for persistent or recurrent nummular adenoviral keratoconjunctivitis (AK) has little benefit because of the frequent recurrences, and mostly "offers" only serious steroid side effects. Since January 1995, we have treated different patients with nummuli after AK with topical Ciclosporin A (CSA) in an attempt to achieve at least the same symptomatic effect as with steroids, however, without side effects. Here, we report about our experiences in a very severe case with longterm treatment. CASE REPORT: The patient was sent to our clinic 4 months after AK with confluent nummuli and Descemet folds, more severe in the right than in the left eye. Best corrected visual acuity was 0.05 in the right and 0.5 in the left eye. Topical CSA 2% 4 times daily was first administered only in the right eye. When after 6 weeks a reduction of nummuli was noted in the right eye, the left eye, which had not improved, was started on the same regime. Therapy was tapered and finally stopped after 12 months in the right and 10 months in the left eye, when only minor changes were left in the corneae. A prompt recurrence of nummuli in both eyes within 4 weeks forced us to resume CSA therapy. At present, both corneae are clear with full vision, and this result is stable with 1 drop of CSA daily. No side effects of CSA therapy have been noted. CONCLUSIONS: The disappearance of nummuli with topical CSA and even more the reappearance of nummuli after cessation of CSA therapy show that topical CSA is about as effective as topical steroids in the symptomatic treatment of non scarred nummuli after KE without the serious steroid side effects. Topical CSA treatment of nummuli after KE is, therefore, a very recommendable alternative for the potentially dangerous steroid therapy. Generally valid data on risk of recurrences, dosage and general effectiveness could only be learned from prospective studies with large numbers of AK patients, which, however, are not available outside epidemics. PMID- 9206750 TI - [Infra-saccular lacrimal duct stenosis caused by bone thickening of the inner nasal bones in tuberous sclerosis]. AB - BACKGROUND: Patients with tuberous sclerosis (Morbus Bourneville-Pringle) may show hamartomatous tumors of different organ systems. 50% of patients display astrocytic hamartomas of the retina. The skeletal system is affected in 40%: peri , en- or exostoses, or cysts may occur. To the best of our knowledge, there are only three descriptions of an involvement of the facial skeleton in the literature. PATIENT: A 54-year-old man presented with recurrent dacryocystitis and lacrimal duct obstruction. Tuberous sclerosis and epilepsy were present since childhood. Besides typical skin lesions and intracranial calcified astrocytomas a retinal astrocytoma was detected in his left eye. Coronal CT scan revealed endonasal bone thickening involving the nasal floor, conchae and lateral walls. Evidence of maxillary and ethmoidal sinusitis was also present. CONCLUSION: The endonasal findings in our patient are most likely a manifestation of tuberous sclerosis of the facial skeleton. They may have favored the development of marked sinusitis and lacrimal duct obstruction. PMID- 9206751 TI - [Insufficiency of a frontalis loop]. AB - After 13.5 years in vivo a 2-0 Terylene frontalis loop was excised because of an insufficiency. Histology showed a persisting giant cell reaction while lymphocytes were almost totally absent. The fibrils of the suture seemed to be intact. However, they were separated by ingrowing connective tissue which, on theoretical grounds, should improve the function of the frontal muscle on the upper eyelid. Therefore, insufficiency of the loop was probably caused by the (subclinical) foreign body reaction or the long-lasting mechanical stress leading to tissue damage. PMID- 9206752 TI - [Release of protein into the extracellular space as a nonspecific response to stress in Escherichia coli]. AB - This paper is concerned with the kinetics of excretion of fluorescent tryptophan containing proteins from Escherichia coli cells kept in physiological saline at room temperature or incubated at 42, 48, and 55 degrees C. The kinetic curves of the extracellular concentration of protein can be described by parameters T and C, where C is the stationary extracellular concentration of the protein and T is the time in which the given concentration is reached. T was found to be a variable, and C was a constant independent of the type and strength of the stress. During the protein release, the viability of the cells was maintained at the initial level, but, after the concentration of the protein reached a stationary value, the culture cells died exponentially. All this allows the protein release into extracellular medium to be considered as a nonspecific response of E. coli to stress. The protein excretion was analyzed with reference to the data on the kinetics of release of other UV-absorbing compounds from the cells. PMID- 9206753 TI - [Regulated expression of bacterial luminescence genes cloned in a multicopy recombinant plasmid]. AB - Luminescence and growth responses of the recombinant strain Escherichia coli Z905 (AprLux+) to different concentrations of ampicillin depended on the source of carbon and energy. When glycerol was used as the substrate, the intensity of luminescence rose with the ampicillin concentration in the medium. Glucose caused catabolite repression of cell luminescence up to the late stationary phase, and ampicillin enhanced this effect. The feasibility of controlling the expression of cloned lux genes was shown. PMID- 9206754 TI - [Morphologic characterization and protein analysis of phages isolated from type strains of Bacillus thuringiensis]. AB - The lysogeny of eleven type strains of Bacillus thuringiensis was studied. Eight new phages were isolated from the variants B. thuringiensis var. tochigiensis, yunnanensis, colmeri, shandongiensis, neoleonensis, silo, mexicanensis, and toguchini belonging to the B1 morphotype. The phages that were isolated from B. thuringiensis var. tochigiensis, yunnanensis, shandongiensis, and mexicanensis possessed transverse disks at their tails and were classified into one morphological group. The protein patterns of the isolated phages were determined. PMID- 9206755 TI - [Restriction analysis of DNA from phages isolated from type strains of Bacillus thuringiensis]. AB - Comparative analysis of the DNA of 11 phages isolated from type strains of Bacillus thuringiensis was performed by means of digestion with restriction nucleases. According to the results obtained, the studied phages were classified into five groups. Within each group, complete identity was revealed in both the restriction fragment number and the molecular mass of individual fragments. No homology was observed between genome structures of phages assigned to different groups. PMID- 9206756 TI - [EEG anomalies in the prodromic phase of Rasmussen's syndrome. Report of two cases]. AB - Electroencephalographic (EEG) recordings were studied at disease onset in two subjects presenting with Rasmussen's syndrome. Particular attention was paid to abnormalities detected during the prodromic phase before clinical outcome suggested the existence of chronic encephalitis. EEG recordings showed focal, polymorphic abnormalities associated with slow biphasic complexes (SBC). These complexes that are composed of two slow waves with opposite polarity, a 150- to 250-mV peak-to-peak amplitude and a 500-ms duration have only been described in inflammatory syndromes of the central nervous system. Their occurrence at onset of Rasmussen's syndrome are discussed. PMID- 9206757 TI - [Somatosensory evoked potentials in rheumatoid polyarthritis with radiologic involvement of the cervical spine]. AB - Somatosensory evoked potentials (SEP) have been recorded in 11 patients with cervical spine involvement, with or without signs of myelopathy due to rheumatoid arthritis (RA). In three patients, SEP have been recorded both before and after cervical spine surgery. In seven cases, the P14 (particularly the P9/P14 amplitude ratio) or P30 potentials were abnormal, whereas other potentials and conduction times were less often modified. Vertebral luxation sites that were predominantly observed at the upper cervical level account for these findings, thus supporting the diagnostic utility of P14 and P30 potentials which respectively take origin in the lower brain stem, close to or into the nuclei cuneatus and gracilis. Postoperative SEP were strongly correlated with the surgical outcome. SEP could be abnormal in the absence of overt clinical myelopathy or vertebral luxations, thus revealing infraclinical damage to the somatosensory pathways. This suggests that SEP recording is useful to discriminate RA patients with upper cervical cord dysfunction from those in whom vertebral lesion proves to have no direct impact on somatosensory conduction. PMID- 9206758 TI - [Penile neuropathy: clinical and electrophysiologic study. Report of 186 cases]. AB - Penile neuropathy is a common disease due to lesion of the sensory branch of the pudendal nerve, ie, the dorsal nerve of the penis. Sexual disorders (deterioration of erection) and sensory signs (hypoesthesia or paresthesia of the penis) are noted in patients with penile neuropathy. Electrophysiological recordings help guide the diagnosis (reduction of the sensory velocity of the dorsal nerve of the penis). Many etiologies can be found (traumatic, toxic, compressive), but the most common lesion is neuropathy related to diabetes. PMID- 9206759 TI - [Visual, auditory and somatosensory potentials in the diagnosis of vitamin B12 deficiency]. AB - We describe visual, brain stem auditory, and somatosensory evoked (VEP, BAEP, SEP) in a 49-year old male patient presenting with subacute degeneration of the spinal cord due to vitamin B12 deficiency. Neurological signs included tetraplegia with a C4-C5 spinal cord compression that was unchanged after surgical decompression. Before treatment, the duration of the bilateral VEP was slightly increased, though their amplitude and morphology were not modified. BAEP were normal. However, abnormalities of SEP with loss of cortical potentials were noticed. Two months after initiation of the treatment, both VEP and SEP recorded in response to median nerve stimulation had improved, but there was still no cortical response to tibial nerve stimulation. Eighteen months later, VEP were normal and recovery of SEP in response to tibial nerve stimulation was observed; however, alterations of peripheral sensory and motor action potentials were still present. These findings are in good agreement with previously reported pathological changes in patients presenting with subacute combined degeneration. Similar abnormalities have been described in patients with multiple sclerosis. Evoked potentials in this case proved to be useful for the diagnosis and the evaluation of the efficacy of the treatment. These findings also suggest that demyelination of the posterior part of the spinal cord and peripheral axonal degeneration might be the main pathological changes related to vitamin B12 deficiency. The former, but not like the latter, were clearly responsive to the treatment. PMID- 9206760 TI - [Distal nerve compression of the leg. Clinical and electrophysiologic study]. AB - Rare distal compressions of lower limb nerves include tarsal tunnel syndrome, entrapment of the first branch of the lateral plantar and medial calcaneal nerves, interdigital neuroma, compression of the deep peroneal nerve on the dorsum of the foot, entrapment of the superficial peroneal and sural nerves. Nerve conduction and electromyographic studies are essential to evaluate these peripheral nerve injuries in order to differentiate focal lower extremity nerve entrapments from ischemic mononeuropathies, lumbar radiculopathies or plexopathies, and generalized peripheral neuropathies. This review summarizes the clinical and electrophysiological findings for each of these rare entrapment syndromes and provides the necessary clues to obtain a correct differential diagnosis with other more common causes of foot and ankle pain and paresthesias. PMID- 9206761 TI - [Modification of in vitro insemination techniques in the treatment of severe male factors in assisted reproduction]. AB - The aim of this study is to detect qualified in vitro insemination techniques in the treatment of the severe oligoasthenotheratospermia which is defined as total motile count in the pretreatment samples (< 5 x 10(6) with > 50% of abnormal morphology). These patients have taken part in the in vitro insemination program of the Assisted Reproduction Unit of the 2nd Department of Obstetrics and Gynecology at Rome University "La Sapienza" during a period between June and December 1995. Several modifications of the standard in vitro techniques have been developed such as: mechanical decumulation of the oocytes, reduction of the volume of culture medium, increase of spermatozoa and oocyte concentration at the moment of insemination. A good fertilization rate was achieved (33%) as regard to the semen sample and procedures utilized. Twelve Ets were performed and 4 clinical pregnancies (25% per patients and 33% per transfer) were achieved. These data demonstrate that by the modification of standard laboratory methods for in vitro insemination, a good fertilization rate and a high clinical pregnancy rate can be achieved in cases of severe male factor infertility without having to resort to micromanipulation techniques. PMID- 9206762 TI - [Early diagnosis of embryo-fetal malformations by transvaginal echography in a high-risk population]. AB - BACKGROUND: High resolution transvaginal sonography permits, in respect of traditional transabdominal scan, an earlier diagnosis of some fetal anomalies and malformations. METHODS: In our prospective study, between 11th to 16th weeks gestation, 820 pregnant patients at high risk for chromosomopathies and for fetal malformations were scanned in order to obtain a detailed survey of embryofetal structures and organs and an earlier diagnosis of fetal malformations. RESULTS: Thirty-two fetal anomalies and malformations (3.9%) were detected, and in 4 cases (0.49%) the diagnosis was obtained later in pregnancy. Fetuses with structural malformations were scanned during pregnancy and, if possible, postnatally. We reconsidered the pitfalls in relation to the time of transvaginal scan and the fetal pathology. CONCLUSIONS: En early prenatal diagnosis of fetal anomalies by transvaginal sonography is related to the knowledge of normal embryofetal development and the pathogenesis of malformations. PMID- 9206763 TI - [Early diagnosis of endometrial carcinoma and its precursors in asymptomatic postmenopausal women. Proposal of a diagnostic protocol]. AB - BACKGROUND: Endometrial carcinoma is presently the most frequent malignant tumor of the female genital tract. In most cases it concerns menopausal women mostly affected by chronic hyperestrogenism not balanced by progesterone. Among the useful techniques for early diagnosis in asymptomatic postmenopausal patients, only echography and hysteroscopy have obtained high reliability levels. Particularly the echographic evaluation of the endometrial thickness is the technique on which the screening of asymptomatic patients should concentrate. Nonetheless its employment on so many people would involve too expensive a social cost. METHODS: In the attempt to identify a limited group of patients to undergo the echographic test, the authors have determined, in a group of 330 asymptomatic postmenopausal women, the maturation degree of squamous cells in PAP-Smear executed for the prevention of the cervical carcinoma, in order to select subjects through a suitable estrogenic stimulation. In cases of high maturation degree (16%) and in a compared group of examinations, a progestin test (MAP-Test) has been carried out to verify the presence of possible pathological endometrial proliferation. They have thus identified a small group of asymptomatic women (6%) to undergo transvaginal echography. In this way 1.8% of the examined patients have undergone hysteroscopy with guided biopsy because of a high risk of carcinoma. RESULTS: The prevalence of endometrial carcinoma so detected was 3/1000. CONCLUSIONS: The authors, intend to continue their experience, end by hoping that protocol adopted by them, should become part of a global prevention program for gynecological oncology. PMID- 9206764 TI - [The PAPNET system in cytological rescreening of cervical smears]. AB - BACKGROUND: The Papanicolaou test has reduced mortality from cervical cancer but has not completely eradicated the disease. A reason for the false negative screening errors may be found in one of three steps of the test (obtaining, processing and interpretation of the cervicovaginal smears). The possible false negative primary screening errors may be detected with the help of rescreening. Therefore, an automated quality control by means of a system such as in the PAPNET is necessary. The PAPNET combines the use of algorithmic image analysis with neural networks and allows to conventionally reassess cervicovaginal smears. Nevertheless, the cytopathologist always decides whether to reassess the sample with the optic microscope as suggested by the presence of abnormal cells in the frames selected by the PAPNET system. Therefore the cytopathologist always formulates the conclusive diagnosis. METHODS: In this study, we rescreened 300 cervicovaginal smears. RESULTS: Only 122 smears have been reassessed by the optic microscope. The human papillomavirus infection and the SIL (squamous Intraepithelial Lesion) diagnoses formulated by means of rescreening with the PAPNET have coincided with the primary screening diagnoses. The only exception was a human papillomavirus infection not previously detected. CONCLUSIONS: Our study indicates that the use of the PAPNET system facilitates the quality control measures and has reduced the false negatives. PMID- 9206765 TI - [Endometrial carcinoma: an increasing neoplasm. Screening and early diagnosis: proposal for a protocol]. AB - Endometrial cancer is the most common cancer of the genital tract and it represents 10% of all cancers diagnosed in women. A protocol for screening and early diagnosis of this tumor has been designed by the authors. All asymptomatic women with risk factors undergo transvaginal sonography. The value of endometrial thickness suggests the need for endometrial sampling. On the contrary, such a procedure is always combined with transvaginal sonography in case of women with atypical genital bleeding. The aim of the protocol is to verify the efficacy of transvaginal sonography and endometrial sampling as combined procedures for screening and early diagnosis of endometrial cancer. PMID- 9206766 TI - [Physiopathology of maternal and feto-neonatal thyroid in pregnant women with endemic goiter]. AB - Endemic goitre is a socially important disease in many regions of Italy. In conditions of euthyroidism, the course of pregnancy and perinatal outcome are not burdened by significant complications. It is useful to control thyroid function and to start L-thyroxine therapy in order to avoid any further increase in thyroid size owing to the goitrogenic effect of pregnancy and to avoid transient hypothyroidism and the nodular evolution of goitre. Recent studies appear to indicate a possible physiological role for thyroid hormones in the development of the fetal CNS as early as conception, an additional motive for the administration of thyroxine in order to prevent pathologies caused by thyroid hormone deficiency in utero. PMID- 9206767 TI - [Primary microinvasive adenocarcinoma of the cervix. A clinical case]. AB - In this case the authors describe a patient with primary microinvasive adenocarcinoma of the eso-cervix, with the cervical canal not involved by the pathology, diagnosed by colposcopy followed by biopsy. Before the diagnosis of microinvasive adenocarcinoma the patient was scheduled for electrocautery of the suspicious area due to the fact that previous Pap-smears had always been negative. The authors underline the important role of colposcopy as a diagnostic tool that should be mandatory before any surgical therapy on the cervix and that allowed to diagnose such a rare cervical tumor. PMID- 9206768 TI - [Essential thrombocythemia in pregnancy. A case report and general considerations]. AB - Essential thrombocythemia is a rare disease of unknown etiology characterized by an abnormal increase in the platelet count which cannot be explained by other identifiable causes such as malignancy, infection, chronic inflammatory diseases or other myeloproliferative disorders. It rarely affects people less than 50 years of age and may be associated with hemorrhagic or thrombotic tendencies. A care of pregnancy complicated by essential thrombocythemia treated with aspirin, antiaggregating agent, throughout pregnancy and with hydroxyurea, a platelet lowering drug is reported. Also examine are some pathogenetic and therapeutic aspects of the thrombotic tendency secondary to elevated platelet count in pregnancy. PMID- 9206769 TI - [Centronuclear myopathy and pregnancy. A case report]. AB - A case of a pregnant woman suffering from centronuclear myopathy is described. The patient has successfully carried out the pregnancy and delivery. PMID- 9206770 TI - [Vitamin E in the treatment of pregnancy complicated by uterine myoma]. AB - The authors describe the encouraging results obtained in the treatment of uterine myomas during pregnancy, using vitamin E at a dose of 300 mg times a day, starting the administration from the time of the first examination of the patient, which took place between week 6 and 12 of gestation. A group of 25 women underwent treatment, aged between 25 and 41 years old, of whom 15 were primigravidas and 10 with one or more previous pregnancies, suffering from uterine myomas in pregnancy, and observed between 1986 and 1994. All the pregnancies continued to term and elective cesarean section was performed associated with single or multiple myomectomy. The neonatal outcome was satisfactory in all cases and no collateral effects were observed in either mothers or fetuses. PMID- 9206771 TI - [Use of nomegestrol acetate in the treatment of menstruation disorders. Our experience in 56 cases]. AB - OBJECTIVE: To investigate tolerability and efficacy of nomegestrol acetate, a new 19-nor-progesterone derivative, in the treatment of oligomenorrhea and amenorrhea; to compare this drug with didrogesteron, a widely-used spatial isomer of progesterone. METHODS: 56 women, aged 22 to 50, affected by menstrual disorders or premenstrual syndrome, entered the study. They were divided in two homogeneous groups by randomization. In the first group nomegestrol acetate was administered 5 mg daily per os; in the second group patients were treated by the usual didrogesteron dose, that is 10 mg twice a day per os; both treatments were administered for 10 days from day 16, during three consecutive menstrual cycles. Efficacy, compliance and biological tolerance were evaluated. RESULTS: 24 patients in the group treated by nomegestrol acetate and 22 in the group treated by didrogesteron completed the study. After the first cycle of therapy, nomegestrol acetate showed a higher efficacy in reducing menstrual loss; results are statistically homogeneous in the two groups at the end of the study. There were no reports of pain in both the groups. Evaluation of biological and clinical parameters at the end of the study did not show significant modifications in any subject. CONCLUSIONS: Nomegestrol acetate is an innovative and efficient molecule in the treatment of menstrual disorders. A good response to the drug is evident in a short time and with very low doses; its good compliance, if compared with other progesteron-derivatives, indicates nomegestrol acetate as a first-line therapy for polymenorrhea, olygomenorrhea and premenstrual syndrome. PMID- 9206772 TI - [Liposarcoma: controversies and conceptual problems]. PMID- 9206773 TI - [Human dirofilariasis in Sardinia: 4 new cases. Review of published cases]. AB - Four new cases of Human Dirofilariasis in Sardinia are described: I subconjunctival and 3 subcutaneous. The patients were men in 3 cases and woman in the other one. The age of the subjects varied from 35 to 58 years. The parasite, Dirofilaria repens, was in all the cases a female. From the review of the literature only 3 other cases were reported in the island. Due to the diffuse presence of both the causal agent, D. repens, in dogs and many species of Culicidae that can transmit the infection to man, it is possible that human cases are more common than reported, many cases passing undiagnosed or simply not published. PMID- 9206774 TI - [Incidence of lymphoproliferative diseases in Northern Italy]. AB - INTRODUCTION: In the last 6 years we have observed an increasing number of cases of lymphoma, extranodal cases being the majority over nodal ones. This fact induced us to investigate the incidence of NHL and HD in the population served by the Lecco Hospital, Department of Pathology, as well as the temporal trend from 1990 to 1995. MATERIALS AND METHODS AND DATA: All hospitals and dispensaries located in the surrounding areas of Lecco including the district of Menaggio and Morbegno are dependent on the Department of Pathology of Lecco Hospital. In fact, the surgical pathology of this area with its 272144 inhabitants converges in our department. We investigated the incidence of nodal and extranodal lymphomas in this population in the period between january 1990 and december 1995 while isolating the number of new cases reported in our files. The incidence per age, population over three-years period has been epressed as the number of cases per million population per year (cases per pmp/y). RESULTS: 285 patients aged 20 to 90 years old (mean 62.5) were selected (51% males, 49% females): they presented 141 nodal lymphomas (36 HD and 105 NHL) and 144 extranodal lymphomas of which 57 primary gastric lesions (22 males and 35 females, mean-age 63.5 in the male group (range 38-85) and 59.0 in the female group (range 31-91)). In nodal as in extranodal lymphomas the diffuse large B-cell lymphomas were the most frequent entity diagnosed, furthermore we noted an increase of incidence in all age groups in the last six years for NHL, mostly NHL HG, in contrast to decrease for HD. About gastric lymphomas, 2/3 were diffuse large B-cell lymphoma with or without low grade component, while the remaining were low grade B-cell lymphoma and only three cases were gastric peripheral T-cell lymphoma. Gastric lymphomas show a higher incidence than in other countries. CONCLUSION: These results show an increase of the incidence of non-Hodgkin lymphomas, mainly of high grade, during six years and in all age groups. On the contrary we observe a tendency in reduction for Hodgkin Disease. Primary gastric lymphomas show a greater increase and their incidence has been estimated in 17.5 cases per 100,000 per 5 years. PMID- 9206775 TI - [Early stage Stewart-Treves syndrome: report of 2 cases and review of the literature]. AB - BACKGROUND: Stewart-Treves (S-T) syndrome is a rare from of angiosarcoma occurring as a complication of lymphedema, classically associated with mastectomy and lymph node dissection for breast carcinoma but also occurring in other forms of chronic lymphoedema. Generally S-T syndrome has a very poor prognosis. Recognition in its earliest stages, at least histologically, can be extremely difficult. MATERIALS AND RESULTS: We report two female patients aged 77 and 68 with chronic lymphoedema of the arm complicating for 9 and 8 years respectively ipsilateral mastectomy for breast cancer treated 10 years earlier in both cases. The first developed violaceous macules on the arm and the second presented with a bluish cutaneous nodule on the upper arm. Histologically the first showed lymphangectasia and a dermal proliferation of thin-walled dissecting vessels with only focally slight endothelial atypia ("lymphangiomatosis"), whereas similar architectural features in the second case were associated focally with overt endothelial atypia and micropapillae. These appearances were regarded as pre malignant (lymphangiomatosis) and malignant (angiosarcoma) respectively. At 3 years follow-up the first patient showed no disease progression, whereas the second patient at her second year of follow-up developed additional nodules. DISCUSSION: In reviewing the literature, approximately 400 cases of angiosarcoma associated with lymphoedema have been reported, of which 360 occurred after ipsilateral mastectomy. Previous controversy as to whether such tumors were truly vascular (rather than simply recurrent carcinoma) has been resolved conclusively in favour of endothelial differentiation. Pathogenetically it seems in these cases that chronic lymphoedema histologically characterized by lymphatic dilatation (lymphangectasia) leads first to proliferation of lymphatics (lymphangiomatosis) with possible slight endothelial atypia. Thereafter there is a gradual continuum of increasing endothelial atypia, followed by multilayering, papillae formation and solid sheet-like tumour. Cumulative published data show that lymphangiomatosis in this clinical setting is premalignant, while the presence of moderate to severe endothelial atypia indicates a diagnosis of (lymph) angiosarcoma. Histological distinction between lymphangiomatosis and ("early") well-differentiated angiosarcoma can be difficult but clearly is of great clinical importance. PMID- 9206776 TI - [Immunohistochemical findings in Huntington's Chorea: report of 9 cases]. AB - The results of an immunohistochemical investigation on neostriatum of 9 cases of Huntington's disease are reported. In all cases the typical neuropathological findings were present (striatum atrophy, neuronal degeneration, gliosis). We did investigate on paraffin slides Synaptophysin (SYN), Neurofilament-protein (NF68), GFAP as well as the neuropeptides Met-Enkephalin (MEnk), Substance P (SP), Somatostatin (SS) and Neuropeptide Y (NPY). These neuropeptides, in particular MEnk and SP, are reported to coexist with the inhibitory neurotransmitter GABA in the neurons of basal ganglia. In all cases, GFAP activity was increased. In 7 cases activity of SYN and NF68 was decreased. In 2 cases, however, SYN immunoreactivity was increased; these findings might represent an expression of "regenerative" changes in surviving neurons. The reactions for neuropeptides did disclose, in accordance with the results of former investigators, a decreased activity of MEnk- and SP-neurons, whereas SS- and/or NPY-neurons appeared almost unchanged. PMID- 9206777 TI - [Bulletin of breast disease: results of the 1990-1994 quinquennium]. AB - All mammary lesions diagnosed at the Institute of Pathological Anatomy of the University of Modena have been systematically filed since 1990 and reported in a bulletin, which is issued twice a year and delivered to health operators. So far, 5.188 cases of breast lesions, comprising 1.999 non-neoplastic pathologies, 1.040 benign tumors, 1.943 primary malignant neoplasms and 206 recurrences, have been filed. During the quinquennium 1990-1994, a progressive numerical reduction in diagnoses of non-neoplastic lesions coupled to an increase of benign tumors has been observed, whereas the number of primary malignant tumors has not changed. In particular, a statistically significant increase in diagnoses of carcinoma-in situ and of fibrocystic disease associated with moderate-risk lesions (atypical hyperplasias) has been detected, whereas the number of cases of single fibrocystic disease has decreased. This reduction, however, is not significant. A slight increase of breast carcinomas smaller than 1 cm and 2 cm, coupled to a decrease of those exhibiting dimensions between 2 and 5 cm, has been found. The collection and systematic analysis of cases of mammary lesions appears to represent a useful tool to study the incidence of different breast pathologies in the general populations. It can also be viewed as a simple way to test the reliability of diagnostic methods used for selection of surgical cases. PMID- 9206778 TI - [Carlo Martinotti: the real discoverer of Martinotti's cells]. AB - The cells of Martinotti are unique neurons of the cerebral cortex with ascending axons. Giovanni Martinotti (1857-1928), professor of anatomic pathology at the University of Bologna, has been claimed to be the discoverer of such cells. Nevertheless, no papers specifically concerning neuroanatomy have been found in his curriculum. The authors have been able to establish that Carlo Martinotti (1859-1918), a pupil of Camillo Golgi (1843-1926) was the legitimate discoverer of the nerve cells carrying his own name. PMID- 9206779 TI - [Bone metaplasia in adenomatous intestinal polyp. Report of a case and review of the literature]. AB - We report a case of metaplastic ossification occurring within a tubulovillous adenoma of the large bowel. To our knowledge, only two cases of this process have been previously described. The pathogenesis is unclear, but the phenomenon seems to be a morphologic curiosity with no clinical significance. PMID- 9206780 TI - [Dedifferentiated (sarcomatoid) chordoma in the base of the skull. Report of a case]. AB - We report a case of "dedifferentiated" chordoma occurring in a 31-year-old man and involving the base of the skull. Morphologically, the tumor was characterized by a bifasic pattern (classical chordoma associates to sarcoma-like areas), and by coexpression of epithelial and stromal markers. Because of these traits, we believe this case shows features superimposable to those seen in sarcomatoid carcinoma. PMID- 9206781 TI - [Laryngeal malignant schwannoma in a 9-year-old boy: report of a clinical case]. AB - A case of malignant schwannoma of the larynx in a 9 year old boy is reported. The lesion recurred 29 months later with the same histological pattern. The malignant peripheral nerve sheath tumours of the larynx are rare and are almost exceptional in paediatric age. The Authors discuss the main differential diagnosis of spindle cell paediatric tumors of the laryngeal region. PMID- 9206782 TI - [Malignant fibrous histiocytoma: an entity in the process of extinction?]. PMID- 9206783 TI - [A new gamma-2 herpesvirus, defined as HHV8 or Kaposi's sarcoma herpes virus (KSHV), could be involved in the pathogenesis of Kaposi's sarcoma]. PMID- 9206784 TI - [Sarcoidosis]. PMID- 9206786 TI - [Aortic ring supravalvular stenosis]. PMID- 9206785 TI - [List of diagnoses...]. PMID- 9206787 TI - [Multidisciplinary quality assurance circles--a quality assurance model for child and adolescent psychiatry clinics]. AB - A complex medical field such as Child and Adolescent Psychiatry can only meet the expanding problems of quality management by multi-disciplinary cooperation. The quality control circle is the central element of the quality control process and of further advances to quality management. In a critical experience-report the concepts, practical issues and further questions of a continuing quality management conference at a University hospital are outlined. It concentrates on practical and process-oriented problems that concern the integration of quality management in practice and research. PMID- 9206788 TI - [God is dead--Friedrich Nietzsche's Oedipus complex]. AB - The early psychoanalysts were influenced by the philosophical ideas of FRIEDRICH NIETZSCHE. The work of SIGMUND FREUD was partly based on NIETZSCHE'S literary world. Just before his mental disaster NIETZSCHE radically destroyed once precious ideals. The roots of this provocative development will be elucidated by analyzing NIETZSCHE'S childhood. A main aspect is NIETZSCHES close mother-tie and especially his father's early death. The father-image of NIETZSCHE'S late years is an essential of this paper. PMID- 9206789 TI - [Possibilities of psychoanalytic treatment in child and adolescent psychiatry in France]. AB - This article describes the context of psychoanalytical approaches and their influence upon child and adolescent psychiatry in an historical and organisational perspective. It deals with some current advances and discussions with regard to technical aspects: indication, frame and process as well as extensions of technique, that will raise weaknesses in questions about training in psychotherapy. PMID- 9206790 TI - [Autogenic training in children and adolescents with type 1 diabetes mellitus]. AB - This paper discusses psychosocial influences of diabetes mellitus type 1 on children and young patients. A group of 21 patients, age 9 to 14 years with Diabetes mellitus type 1 attended a course in "Autogenic Training" for a period of 11 weeks. From the multidimensional questionnaire for children (PFK 9-14, SETZ U. RAUSCHE 1976) 15 dimensions of personality and 5 second rank factors were extracted at the beginning and at the end of training and 5 months later. Additionally HbA1-scores were assessed at the beginning and at the end at a 2 month and a 5 month-follow-up. At the beginning of the course only on one of the 15 scales a significant difference could be observed between experimental group and age related normal population. After training 5 scales and one second rank factor showed significant changes. Significant reduction was observed in: "need for aggressive forms of dominance behaviour" "feeling of submission with respects to other:", "emotional lability" and "tendency for dependence on adults". A significantly increased score was observed in the scale measuring "self confidence regarding one's own meaning, decisions and planning ability". The second rank faktor "neuroticism" was significantly reduced. Against expectations there was no reduction in HbA1 scores. At the end of training HbA1 scores even had increased significantly. But this might have been related to the high frequency of infections during this course. In subjective ratings of training evaluation most of the course members and their parents described fewer problems with attention, less test-anxiety and less aggression and nervousness. The results of this prospective pilot-study are discussed in terms of the psychodynamic influence on diabetes. PMID- 9206791 TI - [Dream recall and sleep disorders]. AB - The present study investigated the relationship between dream recall and sleep disorders. The sample comprised 762 patients who were diagnosed in the sleep laboratory. In the course of the examination they completed the sleep questionnaire SF-B (Gortelmeyer 1986). The results showed a heightened dream recall frequency (DRF) in insomniacs and patients with myoclonia. This result as well as the findings in the control group supports the arousal-retrieval model of dream recall (Koulack u. Goodenough 1976) which emphasizes the importance of nocturnal awakenings. However, this model seems only to be valid for males. In females, DRF is mainly influenced by emotional stress which is best explained by the salience hypothesis of Cohen and MacNeilage (1974). They pointed out that intensive dream emotions lead to high recallability of dream experience. The data gives evidence to the hypothesis of Ermann et al. (1993, 1994) which states that reduced DRF in terms of unsuccessful dream work is accompanied by frequent nocturnal awakenings. DRF of patients with sleep apnea syndrome did not differ from DRF in healthy controls. In addition, sleep apnea parameters did not correlate substantially with DRF. The finding that insomniacs reported more negatively toned dreams in comparison to persons who were examined for sleep apnea but did not showed a pathological apnea index. This may be an hint to increased emotional stress in this patient group. To summarize, the results are promising in clarifying the relationship between sleep disorders and dream life. The next step is to investigate dream reports of these patients by means of content analysis. PMID- 9206792 TI - [Everyday stress and ulcerative colitis]. PMID- 9206793 TI - [Eros and thanatos in the life and work of Heinrich Heine--a study on the occasion of the 200th birthday of the poet]. AB - Themes of love and death in combination determine Heinrich Heines early lyrics. Heine's psychosexual development ist analyzed by emphasising the female influence. His fantasies during adolescence, masochistic tendencies and his inclination to duels are highlighted by referring to Freud's concept of eros and thanatos. PMID- 9206794 TI - [Ernst Simmel and psychosomatic medicine today]. AB - The contribution made by Ernst Simmel to the development of psychoanalytic psychosomatics has been suitably appreciated only during the last few years. At the end of World War I, Simmel attracted the first time public attention with his book on war neuroses and psychic trauma. In 1921 he founded the first psychoanalytic outpatient service, together with Max Eitingon and in 1927 he opened the first psychoanalytic clinic, both in Berlin. He treated patients who were inappropriate for out-patient settings, like psychotic and addicted persons. His own treatment system already embodied essentials of modern clinical psychotherapy, for example the inclusion of nurses in psychotherapy. His clinic served as a model for the Menninger Clinic in Topeka. After the failure of his project following the breakdown of world economy in 1931 and his emigration, his model appeared to have faded out without establishing a tradition. However, further research may assess his possible indirect influence on the German development after the 2nd World War, by reimportation of his ideas from the U.S.A. PMID- 9206795 TI - [Psychoanalytic treatment in the clinic. 1927]. PMID- 9206796 TI - [Reviews in the Psychotherapie Psychosomatik Medizinische Psychologie Journal]. PMID- 9206797 TI - [Health related quality of life and subjective health. Overview of the status of research for new evaluation criteria in medicine]. AB - Health-related quality of life is increasingly accepted as a relevant endpoint in medicine. Available work pertains to development and psychometric testing of measurement instruments. Theoretical papers are largely missing. However, inclusion of instruments in clinical trials has just begun, especially in oncology and cardiology. Based on a review of available literature, areas of applying of quality of life research are described. Foci of attention are conceptual, methodological and practical challenges, as well as the contribution of quality of life research to assessing the results of medical treatment. PMID- 9206798 TI - [Combination of methods and integration of methods as standard in psychotherapy?]. AB - Psychotherapy has undergone a relatively brief but nonetheless dramatic development, but this resulted in an enormous variety, extensive diversity, and good treatment possibilities. Can we say at this stage that the combination and integration of different methods in psychotherapy are today's standards? The authors, a psychoanalyst and a behavioural therapist, consider a greater transparency between the various therapeutic schools as a goal that must still be aimed at before methods can be combined. If such a combination is finally achieved, a standard psychotherapeutic treatment may evolve, since patients suffering from eating disorders, compulsive behaviour or anxiety would profit more from such treatment than from a merely "school"-oriented therapy. Initial ideas for achieving an integration of methods are discussed in respect of inherent limitations and risks. PMID- 9206799 TI - [Trauma diagnosis: difficulty in differentiating between recall and fantasy]. AB - This paper is intended as a contribution to understanding why, up until recently, there have been so few case reports of actual abuse and its sequelae in the psychoanalytic literature. We suggest that psychoanalytic insights into the nature of psychic reality, while indispensable to the evolution of psychoanalytic thinking, have nonetheless had the adverse effect of collapsing any distinction between unconscious fantasies and repressed memories. Moreover, the idea that knowledge of external reality is itself mentally constructed also has diminished interest in uncovering trauma and "real" history. We present a report of an adult analysis that illustrates the recovery of a dissociated memory of sexual abuse that occurred during adolescence, as a springboard to discuss problems analysts have had in dealing with trauma theoretically. We hypothesize that repressed memories and unconscious fantasies can often be distinguished insofar as they may "be stored" or encoded differently, and that consequently the sequelae of trauma and fantasy often, but not always, can be disentangled. We describe some different modes of encoding trauma and some different ways of remembering, re experiencing, and re-enacting it. And, finally, we suggest why traumatic memories are increasingly accessible to patients today. PMID- 9206800 TI - Effect of mifepristone and antiestrogens on uterine PGF2 alpha and PGE2 concentrations in ovariectomized and pregnant rats. AB - Four antiestrogens (anordiol, tamoxifen, RU 39411, ICI 182780) and the antiprogestin, mifepristone (RU 486), were administered to the following three animal models: (1) ovariectomized rats, (2) mated rats treated post-coitally; and (3) pregnant rats treated post-implantation. The antiestrogens were administered alone or in combination with mifepristone at doses effective in preventing and/or terminating pregnancy in rats. The objective of the study was to determine whether these drugs influenced uterine concentrations of prostaglandins (PGF2 alpha and PGE2). Antiestrogens administered alone to ovariectomized rats did not effect uterine PGE2 or PGF2 alpha concentrations; whereas the combination of anordiol/mifepristone increased uterine PGF2 alpha concentration, resulting in an increase in the PGF2 alpha/PGE2 ratio. Mated rats were treated post-coitally for three consecutive days with anordiol, tamoxifen, estradiol and mifepristone alone and with the combination of anordiol/mifepristone and tamoxifen/mifepristone. An increase in uterine PGF2 alpha concentrations and in the PGF2 alpha/PGF2 ratio occurred only in anordiol/mifepristone treated group. A decrease in uterine PGE2 concentrations occurred in animals treated with anordiol, tamoxifen and estradiol, resulting in an increase in the PGF2 alpha/PGE2 ratio. Anordiol (5.0 mg/kg/day) and mifepristone (4.0 mg/kg/day) alone and the combination of anordiol/mifepristone (2.5/1.0 mg/kg/day) administered to pregnant rats on day 7, 8 and 9 of pregnancy induced an increase in PGF2 alpha levels without affecting uterine PGE2 concentration. The changes in PGF2 alpha concentrations induced by anordiol and the combination of anordiol/mifepristone resulted in an increase in the PGF2 alpha/PGE2 ratio. The antiestrogens tested except for ICI 182780 possessed agonist activity when assayed by measuring their capacity to increase the uterine weights in ovariectomized rats. Also, ICI 182789 was the only antiestrogen that did not influence uterine PG concentrations. It can be concluded that ICI 182780 is the only "pure" antiestrogen among those tested. The present results show that antiestrogens and the combination of mifepristone plus anordiol at doses preventing implantation and terminating pregnancy increase uterine PGF2 alpha and/or decrease PGE2 concentrations, resulting in an alteration of PGF2 alpha/PGE2 ratio. These findings suggest that there exists a critical balance of PGF2 alpha to PGE2 concentrations in the uterus required for the normal passage of fertilized ova through the oviduct, initiating implantation of the blastocysts, development of embryos, and maintenance of pregnancy. PMID- 9206802 TI - [Complement in inflammation]. PMID- 9206801 TI - Distribution of prostaglandin E receptors in the rat gastrointestinal tract. AB - AIMS: In order to study the role of prostaglandin in the regulation of the gastrointestinal functions, gene expression of prostaglandin receptors along the rat gastrointestinal tracts were investigated. METHODS: Rats were used for the study. The combination of counterflow elutriation separation of mucosal cells and Northern blot analysis was used to detect the gene expression of prostaglandin receptors in gastrointestinal tracts. RESULTS: In small intestine and colon, prostaglandin E2 EP1 and EP3 receptor mRNAs were mainly localized in the deeper intestinal wall containing muscle layers. EP4 receptor gene expression, on the other hand, was detected in the intestinal mucosal layer. In the stomach, EP1 mRNA was detected in gastric muscle layers, whereas EP3 and EP4 receptor gene expression was mainly present in the gastric mucosal layer containing epithelial cells. In gastric epithelial cells, parietal cells were found to have both EP3 and EP4 receptors. At lower concentrations, prostaglandin E2 inhibited gastric acid secretion by parietal cells probably through EP4 receptors. At higher concentrations, however, it stimulated it. On the other hand, mucous cells possessed only EP4 receptor mRNA. CONCLUSIONS: Thus, it is suggested that prostaglandin E2 modulates gastrointestinal functions through at least three different prostaglandin receptors (EP1, EP3, and EP4), each of which has a distinct contribution in the gastrointestinal tract. PMID- 9206803 TI - [Complement evaluation in the laboratory]. PMID- 9206804 TI - [Deficiencies of C1 inhibitor: hereditary and acquired angioedema]. PMID- 9206805 TI - [Immune and nonimmune complement activation]. PMID- 9206806 TI - [Complement in paroxysmal nocturnal hemoglobinuria]. PMID- 9206807 TI - [Complement in kidney diseases]. PMID- 9206808 TI - [Complement in organ xenotransplantation]. PMID- 9206809 TI - [The interaction between the complement system and immune complexes]. PMID- 9206810 TI - [Letter to a young physician. Is there competition between technology and positive communication with the patient?]. PMID- 9206811 TI - [Clinical uses of flow cytometry in hematological oncology]. AB - In the last decades, the classification of schemes of haematological malignancies have undergone considerable changes both in terms of modifications of previous concepts and of methodological approaches, in parallel with the acquisition of new information on the physiopathological and functional pattern of haemic cells and of their precursors both at the lymph node and bone marrow level. The cyto morphological aspects of haemic were better defined and integrated by the application of cyto- and histochemical methods, which were subsequently supplemented by bioenzymatic and cytogenetic techniques, then by immunophenotypical studies and finally by biomolecular investigations. Through the use of monoclonal antibodies and the introduction both in research and routine diagnostic practice of multiparameter analysis techniques, it is now possible to correlate several cellular parameters, to identify clonality of malignant cells as well as their lineage assignment and maturation stage. Flow cytometry has become an important, rapid and objective method for the diagnosis of haematological neoplasias. In the present survey we have illustrated the different expression of surface, cytoplasmic and nuclear antigens in haematological malignancies, their correlation with the clinical course of the disease and their diagnostic and prognostic significance. PMID- 9206812 TI - [Primary extranodal non-Hodgkin's lymphoma of muscle tissue]. AB - Primary muscular involvement is extremely rare in non-Hodgkin's lymphomas. To our knowledge few cases are reported in literature and all of them concern patients with unifocal muscular lymphoid masses usually growing in one of the extremities. Our case-report, instead, regards a 78 years-old woman presenting primary multifocal muscular involvement by extranodal non-Hodgkin's lymphoma (right upper and lower limbs affected at the same time). Therefore, in contrast with the therapeutic approach suggested by other Authors in such neoplasms (radiotherapy or combined radio-chemotherapy), we preferred to administer only chemotherapy. The treatment led to a complete regression of all lymphoid masses. By now the woman is healthy and disease-free as confirmed at the one-year haematological follow-up. PMID- 9206813 TI - [Primary extranodal non-Hodgkin's lymphoma of the head and neck]. AB - Primitive extranodal head and neck non-Hodgkin's lymphoma represents 10% of total non-Hodgkin's Lymphomas and 5% of total head and neck malignant tumors, preferably 55-65 years old males. The aim of this study was to review the literature and to compare the available data with our cases, particularly referring to the results of therapy. We studied 7 cases of primitive extranodal head and neck non-Hodgkin's lymphoma (5 male and 2 females, mean age 58 years) observed between 1989 and 1994. All patients were treated with polychemotherapy, 2 of them with combined therapy. After a mean follow-up of 44 months, 6 patients (85.7%) still are in complete remission. Primary extranodal head and neck non Hodgkin's lymphomas present peculiar clinical features compared to other lymphomas. The best treatment is the polychemotherapy (including anthracycline) associated (combined) with radiotherapy. A minimal follow-up of 5 years is required, also considering nodal and extranodal relapses not adjacent to the beginning site. PMID- 9206814 TI - [Primary non-Hodgkin's lymphoma of the intestine associated with asymptomatic celiac disease in adults]. AB - Celiac disease (CD), a gluten-induced enteropathy, is characterized by typical intestinal involvement with classical clinic features in childhood and less frequent features in adult patients. Recognizing pauci- and asymptomatic patients is a critical point in the clinical management of CD because of the high mortality associated with the onset of complications. Among these, malignant diseases are the most severe, particularly squamous cell carcinoma and lymphoma, the latter accounting for 50% of all malignancies occurring in CD patients. The authors describe a 57 years old patient with CD and Enteropathy-Associated-T-Cell Lymphoma, who had no intestinal symptoms but only severe pruritus and hypereosinophilia. PMID- 9206815 TI - [Regression of primary gastric lymphoma after treatment with ranitidine]. AB - In a 60 years old patient affected by primary gastric MALT lymphoma stage IE, we observed a complete regression after aspecific treatment with ranitidine 150 mg x 2/day; after regression we treated the patient with anti-Helicobacter pylori therapy for 30 days. Nowadays, after 3 years of clinical and endoscopical follow up we observe no relapse of disease. The elimination of antigenic stimulation of Helicobacter pylori, stopping T-lymphocytes' effect on B-lymphocytes, stopped neoplastic proliferation, causing the complete regression of neoplastic lesion. This clinical case, according to biological data about primary gastric lymphomas, suggests different ways of treatment for primary gastric lymphomas at the lowest stage of disease. PMID- 9206816 TI - [Prognostic factors in breast carcinoma with negative axillary lymph nodes]. AB - Conventionally, tumor size, axillary lymph nodes status, histologic type and grading, proliferative activity, steroid receptors have been used to predict the natural history of breast cancer. In node-negative patients with breast cancer it is most important to identify biological markers that can predict the risk of systemic relapse. These features have been used to allow selection of the best treatment. In this paper we describe the prognostic significance of new tumoral markers in breast cancer patients without axillary involvement. We analyze the prognostic role and the correlation with response to treatment of these parameters: ploidy, oncogenes, p53, epidermal growth factor receptor and cathepsin D. PMID- 9206818 TI - FDA under siege: the public at risk. PMID- 9206817 TI - [Therapy of multiple myeloma]. AB - The association of melphalan and prednisone, introduced in the sixties, allowed a dramatic improvement in the prognosis of multiple myeloma. The subsequent evaluation of different polychemotherapeutic schedules did not ameliorate the results with respect to the melphalan-prednisone association, which remains the treatment of choice in patients older than 60-65 years. In younger patients high dose therapy allowed, in the recent years, significant improvement in terms of reduction of tumor mass and survival. The use of interferon as maintenance treatment allowed a prolongation of the response phase obtained after induction treatment. In this paper we discuss current trends in the management of multiple myeloma and related complication. PMID- 9206819 TI - "Misleading" molecules? PMID- 9206820 TI - Treating AIDS dementia. PMID- 9206821 TI - Testing the power of prayer. PMID- 9206822 TI - Testing the power of prayer. PMID- 9206823 TI - Testing the power of prayer. PMID- 9206824 TI - Metabolic rates. PMID- 9206825 TI - Clinton urges outlawing human cloning. PMID- 9206827 TI - New lead to safer marrow transplants. PMID- 9206826 TI - New clues to asthma therapies. PMID- 9206828 TI - The origin of dogs: running with the wolves. PMID- 9206829 TI - Evolutionary chemistry: getting there from here. PMID- 9206830 TI - Mariner sails into Leishmania. PMID- 9206831 TI - Dealing with database explosion: a cautionary note. PMID- 9206832 TI - Smoky skies, mosquitoes, and disease. PMID- 9206833 TI - Nannobacteria: size limits and evidence. PMID- 9206834 TI - Nannobacteria: size limits and evidence. PMID- 9206835 TI - Nannobacteria: size limits and evidence. PMID- 9206836 TI - Study shows one-fifth of female bison infected. PMID- 9206837 TI - Bison study marks radical shift for research council. PMID- 9206838 TI - Causing cancer by remote control? PMID- 9206839 TI - Does a common virus give HIV a helping hand? PMID- 9206840 TI - Whose finger is on the switch? PMID- 9206841 TI - Human groups as units of selection. PMID- 9206842 TI - Population biology of lymphocytes. PMID- 9206843 TI - A magnetic attraction to high-throughput genomics. PMID- 9206844 TI - A cell growth switch. PMID- 9206845 TI - Xenotransplanters turn xenovirologists. PMID- 9206846 TI - How to use Usenet. PMID- 9206847 TI - 9th National Psychiatry Congress - Psychiatry 2000. The Lost City, 1-4 October 1996. PMID- 9206848 TI - Quality medical care in South Africa. PMID- 9206849 TI - [Managed health care - who will be the winners?]. PMID- 9206850 TI - [Roller skating accidents. A dangerous game on the road]. PMID- 9206851 TI - [Prevention of postoperative thrombosis]. PMID- 9206852 TI - [Intracranial magnetic resonance diagnosis]. PMID- 9206853 TI - [A gynecologist/obstetrician on the Internet]. PMID- 9206854 TI - [Awareness anesthesia--an anesthesiologic dilemma]. AB - Awareness is a traumatic unintentional event during general anaesthesia. Awareness during anaesthesia is a condition which ranges from a state of alertness to deep anaesthesia with partly preserved senses. Awareness can be divided into four groups, depending on the state of alertness and accessibility of the memories after anaesthesia. Diagnostics and follow-up is complicated when awareness with an impaired recall occurs. We conclude that the incidence of awareness has decreased during the last three decades from approximately 1.5% to 0.2% during general surgery. In clinical practice there is no method for the detection of depth of anaesthesia and no anaesthetic technique that prevents awareness during anaesthesia. Guidelines to further reduce the incidence are suggested. Instant treatment of the condition may relieve the acute trauma and reduce the sequelae. We suggest a postoperative visit to ensure quality development for recording and treatment of awareness during anaesthesia. PMID- 9206855 TI - [Atraumatic avascular necrosis of the femoral head in adults. Diagnosis and treatment]. AB - Atraumatic avascular necrosis of the femoral head is a well-known disease which in the majority of cases is associated with conditions like alcohol abuse or steroid therapy. Early diagnosis is important, in particular in younger patients, in order to preserve the femoral head. Plain radiography, computed tomography, scintigraphy, and magnetic resonance imaging are the radiological methods used for detection of avascular necrosis, and especially magnetic resonance imaging has like scintigraphy been found to be well suited for diagnosis in early stages. Characteristic magnetic resonance scan patterns are described. Surgical treatment is necessary in most cases. Various methods are used, according to the stage of the disease. PMID- 9206856 TI - [Roller skating injuries]. AB - The aim of this study was to investigate the number and types of accidents which occur in connection with roller-skates, and also to find out in which anatomical area the most injuries occurred. During the period 01.02.1995 to 31.08.1996, 389 patients sustained injuries in connection with roller-skates. Fifty-nine percent of all accidents happened on a public road. Out of 389 injuries, 174 sustained a fracture: 68% were forearm or distal radius/ulna fractures; 89% were upper extremity fractures. Thirty-three patients were admitted: four for observation due to concussion; 15 for reduction of fractures and application of plaster; and ten for osteosynthesis. Roller-skating accidents are extremely common. Possible prophylaxis includes protective equipment and restricted rinks for roller-skate enthusiast. PMID- 9206858 TI - [Prehospital emergency treatment in the county of Northern Jutland 1993]. AB - As part of the community planning of prehospital treatment in Northern Jutland, a mixed rural and urban region with a population of 500,000 people, we estimated the prehospital treatment in connection with 1617 emergency ambulance services. The investigation included data collected by the ambulance staff, the hospital doctors and review of medical records. The frequency of use of the emergency ambulance per 1000 population was 41 annually, and rapid emergency transport using signals constitutes 65% of call-outs. In the rural region the time from ambulance call until the ambulance reached the patients was longer than in the urban region. The ambulance staff as well as the doctor at the hospital considered a minor number of the patients as seriously ill. In 60% of the emergency ambulance transports the ambulance staff performed therapeutic efforts, frequently oxygen treatment (34%). In 305 transports the staff would have liked to carry out other forms of prehospital service, most often electrocardiographic monitoring and treatment of pain. Only a few patients had need of advanced prehospital services but political consideration should be given to the possibility of advanced prehospital service in both rural and urban regions. PMID- 9206859 TI - [Clinical results after en block double lung transplantation with direct bronchial revascularization. The first three and a half years' experience in Denmark]. AB - En-bloc double lung transplantation with tracheal anastomosis and direct revascularization of the bronchial arteries to the left internal mammary artery has been carried out in Denmark since June 1992. Forty-seven patients (32 with alfa-1 antitrypsin deficiency, 11 with chronic obstructive pulmonary disease, two with cystic fibrosis and two with primary pulmonary hypertension), 25 men and 22 women, average age 39 years (17-64 years), have received their first double-lung transplant with bronchial artery revascularization. Arteriography of the internal mammary artery and bronchial arteries was performed in 42 (89%) of the patients from 1-150 days after the operation. Successful bronchial artery revascularization was demonstrated arteriographically in 40 patients, in two patients the arteriography failed to show bronchial artery revascularization. Arteriography was not performed in five patients due to early complications and death. Bronchoscopy showed rapid, uncomplicated airway healing in 42 patients. Mucosal necrosis under the tracheal anastomosis was found in three patients, and severe obstructive endobronchial growth of the fungus Aspergillus fumigatus was diagnosed in the last two patients. The one- and two-year survival is 83% (Kaplan Meier). Eleven patients are dead, five due to pulmonary causes and six due to extra-pulmonary causes. Pulmonary function became normal in nearly all surviving patients between three to six months after the transplantation. In conclusion, en bloc double-lung transplantation with bronchial artery vascularization has shown good short-term results, and the one- and two-year survival gives hope that a successful bronchial artery revascularization will improve the long-term survival following lung transplantation. PMID- 9206860 TI - [Long-term prognosis in febrile convulsions with and without prophylaxis]. AB - This is a long-term follow-up of occurrence of epilepsy, neurological, motor, intellectual, cognitive, and scholastic achievements in a cohort of children with febrile convulsions (n = 289), randomized in early childhood to either intermittent prophylaxis (diazepam at fever) or no prophylaxis (diazepam at seizures). At follow-up the two groups were of almost identical age (14.0 vs. 14.1 years), body weight (58.2 vs. 57.2 kg), height (168.2 vs. 167.7 cm) and head circumference (55.9 vs. 56.2 cm). The neurological examination, fine and gross motor development on Stott motor test, intellectual performance on the Wechsler Intelligence Scale for Children verbal IQ (105 vs. 105), performance IQ (114 vs. 111) and full scale IQ (110 vs. 108). cognitive abilities on an neuropsychological test battery, including short and long term, auditory and visual memory, visuomotor tempo, computer reaction time, reading test, scholastic achievements and the occurrence of subsequent epilepsy were also very similar. Children with simple and complex febrile convulsions had the same benign outcome. The long term prognosis in terms of subsequent epilepsy, neurological, motor, intellectual, cognitive, and scholastic ability was not influenced by the type of treatment applied in early childhood. Preventing new febrile convulsions appears no better in the long run than abbreviating them. PMID- 9206862 TI - [Roller skating accidents as a cause of intracranial bleeding]. AB - The number of roller-skating accidents has increased in recent years. A new type of "in line" roller-skates has been introduced to the market. We present two cases of intracranial bleeding resulting from accidents using "in line" skates. In one case the brain injuries were fatal. A helmet could probably have prevented or diminished the injuries. We emphasize that a helmet should be part of the correct protective gear in roller-skating. PMID- 9206861 TI - [Psychological consequences of induced abortion]. AB - One hundred and thirty consecutive women were interviewed about the development of psychological symptoms related to induced abortion two days before and four months after the abortion. Sixty-one (47%) participated in the second interview. Of the 61 women, 52% were psychologically influenced before the abortion to an extent which indicated severe crisis or actual psychiatric illness. Four months after the abortion 13 of these women were still psychologically affected. Furthermore, five women who were not affected before the abortion had developed psychological problems. Among ten of these women (16%) the physiological problems could only be related to the circumstance in connection with the abortion. For a number of women (30%) the abortion had a negative influence on their relationships and their sex lives, whereas other claimed that their relationship had become closer because of their reactions towards the abortions. In spite of these conditions all women indicated that their decision about the abortion had been the correct one under the given circumstances. PMID- 9206863 TI - [Pulmonary catheters--time for evaluation?]. PMID- 9206864 TI - [Increased occurrence of pneumococcal bacteremia in Denmark]. PMID- 9206865 TI - [Lipomas]. PMID- 9206866 TI - [Who should treat lipomas?]. PMID- 9206867 TI - [Lipomas and liposarcomas]. PMID- 9206868 TI - [Amino acids and nomenclature]. PMID- 9206869 TI - [Ovarian cancer and the UL technique]. PMID- 9206870 TI - [Investigation of dementia at a dementia clinic?]. PMID- 9206871 TI - [The randomized trials: a mantra of clinical research?]. PMID- 9206872 TI - [The serum immunoglobulin indices in side effects to Triombrast administration]. AB - The aim of the study was further investigation of immune mechanism underlying side effects of radiocontrast examinations. Various immunoglobulins were measured in the serum of 54 patients exposed to excretory urography. 13 of them had side effects. They exhibited elevated levels of serum IgM and IgE. The findings are discussed in terms of suggested allergic origin of negative triombrast effects. PMID- 9206873 TI - [Changes in the oxygen tension of the kidney cortex in exposure to laser irradiation at different wavelengths (an experimental study)]. AB - Experiments with exposure of the renal cortex to different kinds of laser radiation with measurement of pO2 demonstrated that intravenous UV laser radiation inhibits tissue oxygenation. Subvascular blood exposure to infrared and intravenous one to He-Ne laser are beneficial as such radiations improve oxygenation of the renal cortex. PMID- 9206874 TI - [Electrochemical regeneration of the dialyzing solution]. AB - The system with electrochemical regeneration (ER) of the dialyzing solution functions as effectively as a conventional hemodialysis system: ER eliminates creatinine and potassium ions, is inferior in eliminating urea and non-organic phosphorus but superior to hemodialysis in elimination of middle-size molecular toxins. ER enables continuous purification of dialyzing solution, electrolyzer is a constant element of the regeneration system, produced by electrolyzer sodium hypochlorite warrants the system self-sterilization. ER works without systems of water purification, water communications and dialyzer concentrate that is why the procedure of detoxication can be performed outside specially equipped laboratories. PMID- 9206875 TI - [The tubulointerstitial changes in different clinical and morphological types of chronic glomerulonephritis]. AB - As shown by the data available on 228 patients with chronic glomerulonephritis, tubulointerstitial alterations (TIA) occur primarily in mesangiocapillary glomerulonephritis, focal-segmental glomerular hyalinosis/sclerosis, diffuse fibroplastic glomerulonephritis as well as in active nephritic and nephrotic hypertensive types. In 186 patients with chronic glomerulonephritis chronic renal insufficiency evidencing enhanced progression (EP) if arose within 7 years since the disease onset in the presence of TIA was registered significantly more frequently. TIA patients demonstrated EP as a rule in prognostically unfavorable active nephritic and nephrotic-hypertensive types. The relationship between TIA and EP may be explained by their association with unfavourable clinical types suggesting poor prognosis. PMID- 9206876 TI - [The holmium laser in the endoscopic treatment of ureteral strictures]. AB - The introduction of Ho-YAG laser created new opportunities for treatment of urological diseases. The authors used the unit CTH-10 (wavelength 2.09 mu, energy of impulse 0.7-1.5 J, frequency 1-15 Hz). 7 endoscopic corrections of the strictures were performed in 6 females using cutting and ablation properties of Ho-YAG laser (0.5 J, 5-10 Hz). The patency of the urinary tract was restored by dissection through all layers of the scar tissue in 5 females with strictures 0.5 12 cm. Ablation of the scar tissue was used in 1 patient with stricture of the low ureter 0.3 cm. In all the cases surgery ended with establishment of the ureteral stent. Control examinations showed satisfactory results in 5 patients. In one case nephrectomy was demanded. Better conditions for surgery were provided in the presence of nephrostomy drainage. PMID- 9206877 TI - [The diagnosis and treatment of bladder cancer metastases]. AB - The study included 27 patients with regional and distant metastases of bladder cancer. Regional and distant metastases were detected in 12 (44.4%) and 15 (55.6%) patients, respectively. Basic treatment consisted in chemotherapy (MVAC scheme) which combined with reaferon immunotherapy. Chemoimmunotherapy in combination with radiotherapy was used in 8 patients to relieve pain caused by the metastases. The response was seen in 37% of patients with regional metastases and 22% of patients with distant ones. The former and the latter were followed up for 15.9 and 9.3 months, respectively. PMID- 9206878 TI - [The results of the surgical treatment of bladder cancer patients]. AB - 101 patients with cancer of the bladder were operated in the Moscow Cancer Research Center from 1990 to 1995. Cystectomy with varying urinary bypass was made in 49 patients. 52 patients were subjected to bladder resection. The former developed recurrences in 28.9%, the latter in 62% of the patients. Recurrences after the resections were primarily local. 5-year survival of transient-cell bladder carcinoma patients after cystectomy made up 68.2%, after the resection 80.7%. The authors hold that both operations are applicable, but they have specific indications. PMID- 9206879 TI - [The intravesical administration of BCG vaccine in treating recurrent superficial bladder cancer]. AB - The authors consider general immunological effects of intravesical BCG vaccine as an independent method of immunotherapy of superficial cancer of the bladder (CB). 27 patients (15 males and 12 females, mean age 57 years) with a recurrence of superficial CB (T1-2N0M0) were treated. The patients had undergone transurethral or transvesical bladder resection and combined therapy. Intravesical immunotherapy of CB recurrence with BCG vaccine induced persistent changes in the immune system: stimulate lymphocyte activity and phagocytic activity of neutrophils, T-cell function, normalizes function of endogenic suppressors, increases the amount of IgM and CIC. Repeat courses of the vaccine maintain the above immunological effects for a long time. PMID- 9206880 TI - [The current approach to assessing the degree of risk in performing urological operations]. AB - Upon analysis of all the available classification of urological operative risk the authors conclude that classification proposed by the Moscow Anesthesiology Scientific Society meets current requirements as it allows for the opinion of the surgeon, anesthesiologist and functional diagnosis specialist. PMID- 9206881 TI - [The joint use of hyperbaric oxygenation and EHF therapy in benign prostatic hyperplasia and its combination with chronic prostatitis]. AB - 33 patients with benign prostatic hyperplasia (BPH) were exposed to hyperbaric oxygenation (HBO) and SHF waves. Group 1 patients had no prostatic inflammation, group 2 patients had combination of BPH with chronic prostatitis. Both these modalities are considered as to mechanism of action at different periods of the development of urination disturbances. The combined therapy imposed a positive trend in urination parameters, especially evident in group 2 patients. In them mean urination frequency reduced from 14.8-8.5 times to 6.3, mean urine volume increased from 83.6 to 199 ml. Index I-PSS fell in group 1 by 2 and group 2 by 6.4 scores. L index of quality of life declined by 1.2 and 1.6 scores in groups 1 and 2, respectively. It is inferred that combined use of HBO and SHF therapy is highly effective in urination disorders in patients with BPH suffering from or without chronic prostatitis. PMID- 9206882 TI - [Transurethral radiofrequency thermal destruction as the method of choice in patients with benign prostatic hyperplasia with a high degree of surgical risk and the presence of a cystostoma (or pronounced infravesical obstruction with preserved micturition)]. AB - The authors employed transurethral radiofrequency thermodestruction (TRT) in 27 BPH patients with infravesical obstruction and grave somatic status excluding radical surgery. 17 of them had cystostomic drainage. A single procedure was not long and ran without general anesthesia. In patients able to urinate objective and subjective improvement was recorded, in those with cystostomic drainage suprapubic fistula was eliminated in 10 patients. In some cases TRT produced the results comparable to transurethral resection of the prostate. TRT complications comprise mainly infection and inflammation (11.1%). By efficacy and safety TRT occupies an intermediate position between thermotherapy and prostatic transurethral resection. This method is alternative in BPH patients with cystostoma and severe associated diseases. PMID- 9206883 TI - [A trial of the use pf prostaglandin E1 (Edex, Caverject) for the diagnosis and treatment of erectile dysfunction]. AB - The authors have examined 48 patients aged 17-70 years with erectile disorders. In 15 cases sexual adaptation was achieved after teaching them autoinjections. Pharmacological aid with PGE1 (Edex, Caverject) proved effective for sexual adaptation of patients with erectile disorders. In case of well adjusted dose and strict compliance of the autoinjection regimen the number of complications was minimal. In 50-70-year-olds PGE1 do not cause systemic complications due to pharmacokinetics and endogenic nature of PGE1. This prostaglandin is highly effective in some forms of psychogenic erectile dysfunction especially in expecting failure. For such patients 1 or 2 autoinjections of minimal dose are enough to correct erection. PMID- 9206884 TI - [Cardiovascular system function in patients with chronic kidney failure on hemodialysis]. PMID- 9206885 TI - [The 1st International Consultation on Prostatic Cancer (Monaco, 20-22 June 1996)]. PMID- 9206886 TI - [The prediction of the results of extracorporeal shockwave lithotripsy]. AB - The disease history, objective, x-ray and aggregatometry evidence has been analyzed for 188 nephroureterolithiasis patients to make the prognosis of extracorporeal impulse lithotripsy (EIL). If the concrements presented at x-ray picture as structurally homogeneous, medium-contrast, with even margins, fragmentation occurred after 3-4 sessions in 81% of the cases. Low or high contrast calculi with heterogeneous margins disintegrated after 1-2 EIL sessions. Flat calculi were easier to crush than round ones. A decline or absence of changes on the aggregatometry curves prompted the decision on efficacy of further EIL. The calculi which existed for 6 months maximum were the easiest to crush. The outcome of the previous EIL is also essential for prognosis. Inflammation and obesity worsen EIL results. Variant of EIL regimen and the number of impulses are also prognostically significant. PMID- 9206887 TI - [Study of methods of beta-carotene stabilization in powder concentrates for preparation of beverages]. AB - The data on stabilization of beta-carotene in 0.5% concentrate. The concentration of beta-carotene was analyzed by HPLC. The different available antioxidants allowed for using in food industry were tested as stabilizers. The methods of inserting of stabilizers in ready to use beta-carotene concentrate were developed. PMID- 9206888 TI - [Dietary fibers as radiation protectors]. AB - Chernobyl accident caused incorporation of radioactive element in body of people living in this region. A search of the methods and ways for prevention of incorporation and acceleration of elimination of nuclides is very important. Concentrates of dietary fibers (CDF) were suggested for these aims. CDF were isolated from plants and they contain complex of structural carbohydrates and lignin. It was shown that CDF isolated from lemon peel, beetroot residues after wring out, grapes seeds and other plant resources have a radioprotective properties. Authors conclude that CDF can be used in human nutrition for prevention of incorporation of nuclides. PMID- 9206889 TI - [Effects of pectins with different levels of esterification and food additive Medetopect on colon microflora in rats after lead poisoning and lesions by radioactive isotopes]. PMID- 9206890 TI - [Toxicological and hygienic evaluation of a new fodder additive: liquid mash concentrate]. AB - The results of toxicological evaluation of 'concentrate bardjanoj liquid' (CBL) a new fodder additive are presented. It was shown on one generation of rats that CBL at dose of 1 g and 10 g per 1 kg of body mass had no an embryotoxic, gonadotoxic and teratogenic effects. However long-term intake of CBL during 5 month with diet of rats caused the changes in rat body. It was found increasing of relative weights of kidney, spleen and heart, decreasing of contents of total lipids, phospholipids and cholesterol in rat liver. Also the desquamative enteritis in some cases with ulcers was found. These data did not allow to recommend CBL as fodder additive in studied doses. PMID- 9206891 TI - [Selenium contents in human milk after term and preterm labor and in breast milk substitutes]. AB - Selenium contents in breast milk of Russia's and Vietnam's women after term and preterm childbirth and in breast milk substitutes were studied. The concentration of selenium in breast milk of Russia's women was normal after term and preterm childbirth. The level of selenium in breast milk of Vietnam's women was decreased and could not satisfy completely the need of breasted infants. Selenium contents in babyfood "Malutka" Istra' manufacturing company were lower than those in Sybaj' company. PMID- 9206892 TI - [Use of lactulose in products of children's dietetic and therapeutic nutrition]. PMID- 9206893 TI - [Specialized dietetic products and their differentiated use with prophylactic and therapeutic aim]. PMID- 9206894 TI - [Chronological parameters of carbohydrate utilization in the body]. AB - The glycaemic index (GI) is reviewed as example of chronological parameters of carbohydrate utilisation. A food utilisation depends not only on nutrient composition and physical properties of food but also on body metabolism and physiological status including individual peculiarities, daily rhythm and other chronological characteristics of body connected with energy metabolism and biosynthesis of glycoproteins, phospholipids and nucleic acids. Such analytical approaches are very important for practice of human nutrition and animal feeding and also for critical analysis of "fashion" theories in nutrition. PMID- 9206895 TI - [Effects of salt-softener composition on the contents of toxic elements in processed cheeses]. AB - Contents of toxic elements in salt-smelters using in Russia at manufacturing of processed cheeses and in ready cheeses were studied. The levels of arsenic and lead were 4.4 and 1.2 times higher than allowed levels. Sodium three polyphosphate was found as the resource of toxic element incorporation in processed cheeses. The control of contents of toxic elements in salts are needed at manufacturing of processed cheeses. PMID- 9206896 TI - [Determination of angiogenin in cow's milk based on a competitive test "pancreatic RNAase/placental inhibitor of RNAase--angiogenin"]. AB - Contents of active factor of blood vessel growth angiogenin was analyzed in cow's milk by the method of competitive test in system "pancreatic RNAase/placental inhibitor of RNAase-angiogenin". High level of RNAase in milk limits using this test. To avoid this limitation the method of RNAase elimination from milk was elaborated. PMID- 9206897 TI - [Cranberries: chemical composition, nutritional and medicinal properties]. AB - The contents of protein, fats, carbohydrates (glucose, fructose, sucrose, dietary cellulose), some vitamin (ascorbic acid, riboflavin, beta-carotene, vitamin E), polyphenolic compounds, organic acids, pectin, glycosides, macro- and microelements in red bilberry are presented. Antimicrobial effect and other effect of red bilberry and its use in traditional medicine and dietetics are also outlined. PMID- 9206898 TI - [Ecological certification]. PMID- 9206899 TI - [Today's bread market]. PMID- 9206900 TI - [Beta receptor blockers: renaissance in therapy of heart failure]. PMID- 9206901 TI - [Beta blockers--1997 update]. AB - Beta blockers have been used as first-line drugs in the treatment of numerous cardiovascular disorders such as hypertension and ischemic heart disease, as well as for certain non-cardiovascular diseases for more than 30 years. However, the administration of these safe and effective drugs declined during the 1980s, whereas the use of others such as calcium antagonists and ACE inhibitors increased for these indications, frequently without convincing evidence of any clinical advantages of these agents in hard end points. During the past two or three years there has been a renaissance of beta adrenoceptor antagonists, most probably due to increasing awareness that beta blockers have been shown to reduce morbidity and mortality when compared with other therapeutic agents or placebo in numerous diseases. Furthermore, congestive heart failure has changed from being a contraindication to an indication, and suspected side effects were not confirmed on further investigation. Last but not least the reasonable costs of beta blocking drugs may have become a more important consideration than before. The trend back to beta blockers may also be due to the fact that physicians attach more importance to effects on the hard end points, namely a decrease in morbidity and mortality, than to surrogate end points. According to the present state of the art, beta blockers should be recognized as the drugs of choice, particularly in the treatment of arterial hypertension and coronary artery disease (especially after myocardial infarction), unless contraindications are present or unacceptable side effects occur. Congestive heart failure, peripheral arterial disease (PAD) and diabetes mellitus are no longer considered absolute contraindications. PMID- 9206902 TI - [Brain metastases of colon and rectum carcinomas]. AB - We analyzed 20 cases with brain metastases from colon or rectum carcinoma. Fourteen were treated with radiotherapy alone (total dose 30-60 Gy), six with neurosurgery plus radiotherapy (total dose 30-40 Gy). All patients had advanced primary tumours (T3 and T4), most of which were poorly differentiated; lymph node metastases were common. In 5 patients (25%) the brain was the first site of distant metastases. Ten patients (50%) had a solitary brain metastasis. As a tendency, the results of surgery plus radiotherapy were superior to those of radiotherapy alone, with respect to palliation of symptoms as well as to local tumour remission and survival. Overall median survival was only 51 days. The 1 year survival rate was 6%. In 5 of 14 cases (36%) symptomatic improvement was observed after radiotherapy alone. Partial remission of the brain metastases occurred in 3 of 14 cases (21%). The presence of extracerebral metastases was the most important prognostic factor. Selected patients considered to have a favourable prognosis may profit from combined treatment, i.e. neurosurgery plus radiotherapy. Despite the short survival time, stereotactic irradiation should be evaluated as an alternative to conventional radiotherapy in the remaining patients because the palliative effect of fractionated external irradiation is relatively disappointing. PMID- 9206903 TI - [Rare localization of a monstrous thymoma in the anterior inferior mediastinum]. AB - 90% of all thymomas are localized in the anterior-superior mediastinum, seldom in the inferior or posterior mediastinum. We report the rare case of a 61 year-old female patient who presented with a large thymoma of the anterior-inferior mediastinum with the chief complaint of position-dependent dyspnoea secondary to compression of the left lung. Surgical resection of a tumor weighing 1053 g was successfully performed and 8 months postoperatively the patients is asymptomatic and without evidence of recurrence. PMID- 9206904 TI - [Nephrotoxicity of analgesic combination preparations]. PMID- 9206905 TI - [Comment on the contribution: "Usefulness of fixed analgesic combinations exemplified by thomapyrin"]. PMID- 9206906 TI - [Cytotoxic T-lymphocytes and the recognition of tumor antigens--an approach to "preventive tumor vaccination?"]. AB - In spite of modifications of the intra- and postoperative therapeutic strategy as well as new adjuvant protocols, most malignant diseases have a poor prognosis. This demonstrates the need for new approaches in tumor therapy. One alternative is adoptive immunotherapy (AIT). AIT is based on the observation that cytotoxic T lymphocytes (CTL) may be generated with the ability to lyse autologous tumor cells in vitro and in vivo. Aim of current research activities is to determine specific tumorantigens and the clinical applicability of cell-mediated immunotherapy. The immunobiology of the cell-mediated immune response to cancer with focus on major-histocompatibility complex class I and CD8+ T cells is reviewed. Recent advances in the identification of tumor-associated and tumor specific antigens were summarized. Previously performed clinical studies were reviewed. We examined the implications that the discovery of these antigens might have on the development of new anticancer vaccines. PMID- 9206907 TI - [Personal experiences with emergency coronary artery revascularization after failed PTCA--early and late results]. AB - In recent years the equipment and techniques for percutaneous transluminal coronary angioplasty (PTCA) have been improved and today complex and distal stenoses are also treated in this way. Subsequent to failed PTCA some patients undergo emergency CABG. Between January 1991 and March 1995 3,520 patients have been treated by PTCA in our hospital. 61 patients (1.7%), mean age 61.1 years, underwent subsequent emergency CABG after PTCA. 46% had single-, 33% double and 21% had triple-vessel-disease, the mean left ventricular function (LVEF) was 65%. The mean number of bypass grafts was 1.9. The internal mammary arteries were never used under these emergency conditions. 9 patients (15%) developed perioperative myocardial infarction and in two of them the LVEF decreased under 30%. The hospital mortality was 6.6% (= 4 perioperative deaths). 6 of 61 patients were lost for follow-up; 90% of the hospitals survivors were followed for 244 months (mean 17). During this period there were 4 late deaths (3 cardiac and 1 non-cardiac). Actuarial survival at one year was 90%. 80% of the long-term survivors were in the NYHA functional classes I and II. In patients with double- or triple-vessel-disease PTCA almost always effects an incomplete revascularisation. Emergency CABG following failed PTCA is associated with an increased mortality and morbidity. The long-term prognosis is similar to that of an age-sex matched general population. PMID- 9206908 TI - [Value of intraoperative laparoscopic cholangiography]. AB - Discussion about the necessity of intraoperative cholangiography restarted when laparoscopic cholecystectomy was established. The value of cholangiography was examined in a prospectively randomized study of one hundred patients. We could show that the routinely performed intraoperative cholangiography represents a careful, secure and sensitive method for the detection of common bile duct stones. As it is not very time consuming nor linked to high costs we believe it to be unrenouncible. It allows a detailed anatomic presentation and may be combined with ERCP for definitive treatment of bile duct stones. PMID- 9206910 TI - [Transthoracic-transabdominal resection of esophageal carcinoma]. AB - 54 patients suffering from esophageal cancer have been treated in a period from 1990 to 1994. In 29 cases curative resection was possible, corresponding to a resection rate of 54%. Average age of resected patients was 62 years. According to pTNM-classification the stages T1 and T2 amounted to 45%, T3 and T4 to 55%. Lymphatic node metastases were discovered with an incidence of 55%. In patients treated conservatively more unfavourable stage distributions and increased rates of lymphatic node metastasis were shown. Transthoracal-transabdominal esophageal resection was preferred as curative management. Lethality amounted to 13.8%. In 3 of 4 lethal cases after resection autopsy confirmed absence of tumor. Lethal complications were two respiratory insufficiencies, one suture line dehiscence and one alcoholic delirium. Survival rates were calculated by life-table-method. We consider the transthoracal-transabdominal esophageal resection as an acceptable therapeutic option in esophageal cancer offering a real chance of enduring curing. PMID- 9206909 TI - [Changes in the cytokine concentration (Il-6, Il-8, Il-1ra) and their cellular expression of membrane molecules (CD25, CD30, HLA-DR) after surgical trauma]. AB - Elective surgical approaches and trauma cause changes in the production of different cytokines, an increased production of acute phase protein and changes in the expression of different cell surface markers. METHODS: In a prospective study we examined the C-reactive protein level, the production of the cytokines IL-6, IL-8 and IL-1 RA in 25 laparoscopic and 21 conventional cholecystectomies. In addition the cell surface markers CD25 and CD30 on different cell populations and HLA-DR on monocytes were measured. Statistical analysis was made by Student's t-test and Mann-Whitney's rank sum test. RESULTS: The humoral markers showed a more distinct increase in patients operated on conventionally two and 24 hours after surgery, the differences between the two surgical approaches were significant (p < 0.01). The cell surface markers CD25 and CD30 showed the same reaction. The HLA-DR expression on monocytes was significantly lower in patients operated on conventionally. CONCLUSIONS: Elective surgical approaches cause changes in the immune system, which can be evaluated by the reaction of cytokines and cell surface markers. Laparoscopic cholecystectomies cause less activation of the immune system than conventional operations. PMID- 9206911 TI - [Late results of joint-preserving femur head necrosis surgery]. AB - Early diagnosis of nontraumatic avascular necrosis of the femoral head has been much improved by introduction of magnetic resonance imaging (MRI). Intertrochanteric osteotomies are dominating in operative treatment of advanced necrosis of the femoral head. Studies concerning with long-term results are rare in literature. The aim of the present study was the assessment of the long-term results (> 10 years) of advanced avascular necrosis (n = 33 hips, 8 stage Ficat II, 19 stage Ficat III, 6 stage Ficat IV). In 29 cases of joint-preserving operation was primarily performed. The average follow up time of the patients was 13 years. In 13 patients a joint-destructive operation (total hip replacement or arthrodesis) was secondarily necessary. The average time interval between joint preserving and -destructive operation was 53 months (minimal 10, maximal 180 months). Quality of life is often reduced in patients after joint-preserving hip operations. Only 3 of 14 patients with still existing joint-preserving operation were painless postoperatively and could continue their former job. The other patients suffered from pain and reductions of their activity of daily life. 4 of them had to retire early. PMID- 9206913 TI - [Patient education: what does the patient know, what does he want to know? Thrombosis and wound infection]. AB - The manners of which preoperative informations are given to a patient depend on several different points. So not only the patient's education, his knowledge of medical items and his intelligence are important, but also the juridical situation and the doctor's ability to explain the situation in a way the patient is able to understand. The aim of this study was to evaluate the patient's previous knowledge about two major complications in surgery--thrombosis and wound infection. Beside this it was looked for the importance of confidence in the doctor and previous information about the operation. PATIENTS AND METHOD: 117 patients aged 16 to 87 years were tested anonymous by a standardised questionnaire. RESULTS: There is only little knowledge about the time a thrombosis develops, embolism, a wound infection and their consequences. On the other side, more than 50% of the patients were able to explain correctly the symptoms of thrombosis and the time a wound infection occurs. There was a correlation between the statement of knowledge about thrombosis (p < 0.01) and wound infection (p < 0.01) and the ability of correct explanation. Almost 90% of the patients thought that information about the operation is as important as the confidence in their surgeon. More than two third of the patients wanted to be informed about "everything" that possibly could happen. The main sources of information were neighbours, friends, the press, books and at last the doctor. CONCLUSION: There wasn't any influence of former surgeries, the awareness of complications or education on the active knowledge of the patients. Therefore we think that the way of giving explanation to a patient should help reducing fear of an uncomfortable situation and take account to the patients need for information. Thus, the patient will be able to make a real decision. PMID- 9206912 TI - [Analysis of primary management of bladder malfunction in traumatic paraplegic patients]. AB - Intermittent catheterisation is the demanded therapy of bladder paralysis during the spinal shock. We investigated the realisation of this concept. In 1994 after first treatment in other hospitals 97 patients were treated in the Thuringian Spinal Cord Centre Sulzhayn. The primary treatment of the paralysed bladder was: indwelling catheter: 44 patients (45.4%), suprapubic catheter: 30 patients (30.9%). Only 15 patients (15.5%) were catheterized intermittently. No urological treatment was carried out in seven cases. CONCLUSION: The primary treatment of bladder analysis in spinal cord injured patients has to be improved. PMID- 9206914 TI - [Wound rupture--incisional hernia]. AB - The development of incisional hernias after major laparotomies depends on several factors: preoperative risk factors, constitution of the patient, site of incision, and technique of abdominal closure. There is a strong correlation of postoperative incisional hernias and the number of risk factors in a range of 11 to 44%; transverse or oblique incisions are followed by hernias in a rate of 3.8% in contrast to midline incisions (13.6%). PMID- 9206915 TI - [Acute compartment syndrome of the tibia--complication of popliteal artery aneurysm]. AB - The compartment syndrome is a common complication in the traumatology of the lower limb. In vascular surgery it is observed following emergency revascularisation. It is a rare condition as an initial symptom of a peripheral clotted aneurysm. By means of a case with a complicated popliteal aneurysm the differential diagnosis of the non-traumatologic compartment syndrome is discussed. PMID- 9206916 TI - [Amebic colitis and amebic liver abscess--epidemiology and personal case report]. AB - The large intestine reacts relatively monomorphically to different stimuli. From this differential-diagnostic problems may result. The history of a patient is described which could be pursued clinically over 12 weeks and during the course of which the correction of the diagnosis ulcerative colitis into amoebic colitis was necessary. It is concluded that in every symptomatology of colitis bacterial and parasitologic examinations of the faeces should be performed primarily specially if there is a history of overseas travel. In these cases it must be also thought of spontaneous amoebic infections. PMID- 9206917 TI - [Encounter of the Ernst von Bergmann and Erwin Payrs surgical schools at the Jena University Clinic]. AB - This paper gives a short historical overview on the work of distinguished Professors such as Erich Lexer, Nikolai Guleke, Heinrich Kuntzen and Theo Becker, who worked in the Surgical University Hospital in Jena from 1911 to 1981. The main areas of their clinical and scientific activities are described, as well as the private and human qualities of these well-known personalities. PMID- 9206918 TI - ["Patients with genetic diseases can, by surgical intervention, be sterilized..." Mandatory sterilization in Germany during the era of national socialism]. AB - The ethical problem of permissibility of sterilisation for eugenic reasons is being discussed since the end of the last century. For many years restriction and rejection were the predominant opinions. A bill passed July, 1933--"Gesetz zur Verhutung erbkranken Nachwuchses" ("Act of prevention of heredopathic progeny")- imposed juridical regulations to prevent offspring with inherited diseases in National Socialist's Germany. No major protests had been made against such compulsion put upon the medical profession. Papers dealing with questions of surgical techniques had been published in Zentralblatt fur Gynakologie predominantly in 1935 and 1936. The significance of the scientific analysis of mandatory sterilisations for so-called eugenic reasons performed in almost 400,000 women and men in Germany between 1933 and 1945 has only recently began to be recognised by the scientific community in the last decade addressing predominantly the regional impact of this jurisdiction. PMID- 9206919 TI - [Quality of life in relation to patient education regarding surgical procedures in primary breast carcinoma]. AB - Breast conserving therapy is seen as a profit according to life quality. The patients have been subjected to the surgeon suggestions during decision-making about the primary therapy. The purpose of this study was to examine the relationship between the degree of preoperative information of breast cancer therapy and patients' choices of treatments in cases suited for breast conserving therapy. 138 recurrence free patients after breast conserving therapy (BCT) or mastectomies (ME) due to a breast cancer staged as pT1-2N0-1M0 and missing contraindication to BCT were interviewed using an observer checklist. The degrees of preoperative information, reasons for decision to mastectomy, though BCT was possible, were correlated with the postoperative life quality. Patients decided for BCT were averaging 56.0 + 12.3 years and significantly (p < 0.05) younger than patients decided for ME (60.4 + 10.5), whereas tumor size, nodal status and adjuvant therapies were comparable. The results indicate, that subjects' choice of treatment was unrelated to the amount of information. 87% (BET) respectively 78.3% (ME) patients evaluated, that preoperative information was enough (p = 0.19). The most frequent reasons for preference of mastectomy were the "perception that survival would be diminished if mastectomy was not done" (93.5%), "avoidance of radiotherapy" (60.9%) and "no partner" (34.8%). According to expectation the body image in the BCT-group (84.8%) was significantly (p = 0.0007) more positive than in the ME-group (58.9%). Nevertheless only 5 (10.9%) patients after ME have felt sorry for their earlier decision. There were no significant differences between the two groups with regard to partner- and sexual adjustment as well as physical well-being. Despite being fully informed of treatment possibilities and no medical contraindications to BCT nearly one third preferred mastectomy due to different reasons. When the patient was involved in the clinical decision-making process the mastectomy indicates not generally a loss of life quality, though nearly 40% are dissatisfied with their nude body image. PMID- 9206920 TI - [Possibilities and limits of mammography with special reference to breast carcinoma--a comparison of clinical, mammography and histologic diagnoses]. AB - This paper informs about the diagnostical accuracy of clinical examination and mammography at the women's Hospital of the Stadtische Klinikum Zwickau from 1989 to 1992 when pathological changes of breasts are diagnosed. A total of 508 diagnostical biopsies was performed during this study period. The histological examination resulted in 276 benign breast lesions and in 232 breast cancers. In 432 cases there was a lump palpable in the breast that was noticed at first by more than 60% of women themselves. 288 (56.7%) correct and 220 (43.3%) false diagnoses were established following the clinical examination. This resulted in a sensitivity of 100% with a specificity of 20.3% and an overall accuracy of 56.7%. However, with the help of mammography 386 (76.6%) pathological changes of breasts were found to be correct. 118 mammograms proved to be false. Out of these 90 (17.8%) had a false-positive diagnosis and 28 (5.6%) a false-negative one. The overall accuracy of mammography was 76.6% (sensitivity 87.9%, specificity 67%). PMID- 9206921 TI - [Pelvic exenteration: effects of surgical method on quality of life]. AB - This prospective longitudinal study explores the quality of life of patients who underwent pelvic exenteration (n = 21) with or without reconstructive procedures by standardized questionnaires and semistructured interviews. Quality of life is defined in following categories: physical status, psychosocial issues, medical interaction, marital and sexual problems. At three points in time (preoperatively, 4 and 12 months postoperatively) the quality of life was mostly affected by worries about tumor-progress and not being able to care for oneself. Patients who received reconstructive or preserving procedures felt less restrained in all categories than those without reconstructive surgery although the preoperative situation was not different. PMID- 9206922 TI - [Effectiveness of radical therapy in vulvar carcinoma. An analysis of 148 cases]. AB - 148 patients with primary squamous cell carcinoma of the vulva were treated by surgery at the N. N. Petrov-Cancer Research Institute St. Petersburg. There were 73 unilateral lesions confined to the labium majus or labium minus, 17 to labium majus and minus, 41 to the clitoris and 17 lesions to other structures. 28 patients had FIGO I lesion, 58 a FIGO II, and 62 patients had a FIGO III tumor. In 41 cases the depth of infiltration was 1-5 mm, in 83 cases 6-10 mm, and in 24 cases the depth of infiltration was greater than 11 mm. Radical vulvectomy with inguinal lymphadenectomy was performed in 115 cases, a simple vulvectomy in 33 cases (Nx). In 53 patients lymph nodes were positive and in 62 negative. The patients were followed for at least 60 month and none have been lost to follow up. The stage dependent 5-year-survival rate was 96.4% (FIGO I), 87.7% (FIGO II), 62% (FIGO III), and 79.2% (overall). The 5-year-survival rate of lesions of the labium minus was 94.6%, of the labium majus was 83.1%, of the clitoris--82.2%. A poor prognosis was found in the cases of a tumor involvement in labium majus and minus and/or other vulvar structures (60% and 59.9%). Increasing depth of invasion was associated with decreasing 5-year-survival rate: 97.5% ($ 5 mm), 72.4% (6-10 mm) and 65.4% (> 11 mm). In patients with lymph node involvement the prognosis was significantly better than in those with negative lymph nodes (61.6% vs. 86%). On the other hand, the prognosis of patients with a solitary lymph node metastasis was significantly better than in patients with two or more lymph node metastasis (79.6% vs. 51.6%). Tumor localization, tumor size, lymph node status and especially the depth of invasion are the important prognostic factors in vulvar cancer. PMID- 9206923 TI - [Cardiotocographic changes after umbilical cord puncture and umbilical cord transfusion]. AB - In connection with umbilical cord punctures and transfusions the degree of pathology still tolerable after these procedures as well as the influence on subsequent obstetric management has to be discussed. In a prospective study we evaluated 98 cordocenteses with (n = 44) and without (n = 54) transfusion (25-37 week of gestation). 30-160 ml of blood were transfused. In all cases the umbilical vein was punctured at the point of insertion into the placenta (46x posterior and 52x anterior wall placenta). A CTG was performed prior and after the punction and was evaluated according to the Fischer score. After the procedure, the course of pregnancy was normal in all puncture patients and in 52 of 54 patients who also underwent a transfusion. Because of a pathological CTG due to a severely hydroptic fetus, a caesarean section had to be performed in one woman seven hours after umbilical cord transfusion. In another patient an emergency section was necessary immediately after the umbilical cord transfusion, due to persistent fetal bradycardia. In the puncture group mean Fischer score values decreased from 9.1 to 7.5, in the transfusion. In another patient an emergency section was necessary immediately after the umbilical cord transfusion, due to persistent fetal bradycardia. In the puncture group mean Fischer score values decreased from 9.1 to 7.5, in the transfusion group from 8.6 to 7.4. The results were more unfavorable when amplitude and occurrence of accelerations were considered, especially in the transfusion group. In one fifth of the puncture cases and one fourth of the transfusion patients the criterium of baseline crossings improved after the procedure. In summary, a pathological CTG is to be expected after umbilical cord punctures and transfusions, with however, only the necessity of surveillance. Only in cases of persistent fetal bradycardia an active obstetric management is indicated. PMID- 9206924 TI - [Telecommunication--a medium for improving prenatal diagnosis and gynecologic ultrasound diagnosis? Initial experiences]. AB - To establish the requirements for real-time transfer of an ultrasound examination via telecommunication network the following tests were performed: The ultrasound data were transferred from the video out of an ultrasound system to a basis terminal of the German Telekom. Simultaneously, an external video camera filmed the positioning and movements of the ultrasound transducer, and the verbal comments were recorded. These informations were transmitted to Karlsruhe and London, where they were rerouted to the examination room in Heidelberg. Here the informations were received on a Telecom reception unit/terminal and compared directly with the initial signal. The quality was sufficient if the moving ultrasound images and the camera image of the transducer as well as the oral comment were transmitted over 2 parallel ISDN lines. The delay to a real-time transmission of the examination process is only in the range of milliseconds. If only one ISDN line is used, the image quality is unsatisfactory, three parallel lines do not bring significant improvement of image quality. Telemedicine seems a new possibility to bring the knowledge of specialized centers to the practicing gynaecologists thus avoiding unnecessary referrals. Still unanswered, however, are the problem of liability, data protection and costs. PMID- 9206925 TI - [Parenchymal thoracic endometriosis--diagnosis and therapy]. AB - We report a case of parenchymal pulmonary endometriosis in combination with severe endometriosis genitalis externa. The difficulties on the way to final diagnosis are described, also the different possibilities of hormonal treatment that results in ovarian suppression. In our case the patient is without any symptoms since three years using a GnRH-Agonist as first line therapy and as second line therapy high-dose medroxyprogesterone acetate. Because of the very good clinical results after hormonal suppression surgical intervention might become necessary only in certain cases. This can be the case when after intolerance of hormonal treatment angiography of lung vascular system on the bases of number and localization of endometrial lesions shows good operative conditions. PMID- 9206926 TI - [Diagnosis and follow-up of post-transplantation osteopathies]. AB - The skeletal status of transplant recipients needs routine assessment both before and after organ transplantation for the following questions: 1) is the patient on risk for developing osteoporosis independently of transplantation? 2) are there signs of significant pre-transplantation bone disease? 3) does the patient develop post-transplantation osteopathy? Although biochemical markers of bone metabolism provide useful data to each of these questions, the complex metabolic pertubations seen in chronic organ failure make it mandatory to interpret biochemical findings only within the framework of other clinical tests and imaging techniques. Pre-transplantation osteopathies require specific and often broad clinical and laboratory evaluation, and biochemical tests should be chosen according to the underlying disease and type of organ failure. In this respect, biochemical markers of bone metabolism may provide information on the degree of the skeletal dysbalance. Likewise, biochemical markers of bone turnover may be helpful in monitoring skeletal metabolism after organ transplantation. PMID- 9206927 TI - [Abdominal aortic aneurysm: surgery, indications, technique, outcome]. AB - The fate of a patient with an abdominal aortic aneurysm] (AAA) is influenced by the risk of rupture and embolism. When the indication for operation is considered, individual associated risk factors have to be taken into account. With regard to the literature, the following recommendations concerning indication for surgery can be given: emergency surgery for symptomatic or ruptured aneurysm; elective surgery: aneurysms 5 cm diameter or growing AAA 5 mm/year, patient with acceptable individual risk for operation; asymptomatic aneurysms less than 5 cm in diameter, without growth in patients aged over 75 years and/or considerable perioperative risk should not be operated on: sonography should be done 3-monthly as a continuing control. Finally the results in our institution are presented for elective surgery: 30-day mortality 3.5%, AAA with rupture, no shock: 20%, ruptured AAA with shock 47%, respectively. PMID- 9206928 TI - [Self-injury behavior]. AB - BACKGROUND: Self-damage is defined as intentional injury of the own body. Patients with this disorder often consult and deceive surgeons. In case of factitious disease the diagnosis of self-injurious behavior can be difficult. METHODS: A literature review on self-injuring behavior was done with special emphasis on its clinical presentation in surgical departments, its psychodynamic background and the therapeutic consequences. RESULTS: Self-damaging behavior is most frequent in adolescent females. Both a disturbed relation with the own body and with fellow-beings is the problem of all patients. Deprivation, physical or sexual abuse are common in the biography of these patients. The feelings of internal emptiness and unbearable psychic tension are the immediate psychodynamic causes of the self-damaging act. Psychotherapeutic strategies are aimed at learning to express emotions in a better way, to care for the own body, and to establish confidential and stable relationships. CONCLUSIONS: The knowledge of the psychodynamic background facilitates the therapeutic approach to patients with self-injuring behavior. PMID- 9206929 TI - [Flow cytometry for extensive thoracic diagnosis]. AB - Immunophenotyping of cells by flow cytometry has become a routine test to diagnose pulmonary and mediastinal diseases. Peripheral blood, extravascular fluids, bronchoalveolar lavage (BAL) and suspension of single cells obtained by fine-needle aspiration can be used. Peripheral blood (MOAb for immunophenotyping of lymphocytes: CD14, CD45, CD3, CD19, CD4, CD8, CD16/56, HLA DR, CD38, CD25) is the material of choice for detection and monitoring of immunodeficiences. BAL (MOAb for immunophenotyping of lymphocytes: CD14, CD45, CD3, CD19, CD4, CD8, CD16/56, HLA DR) is used mainly for differential diagnosis of extrinsic allergic alveolitis (low CD4/CD8 ratio) and sarcoidosis (high CD4/CD8 ratio). The enumeration of alveolar macrophage subsets is an important tool to establish diagnosis of histiocytosis X (CD1a > 3%). Extravascular fluids, suspension of single cells and BAL are preferred materials for detection and classification of non-Hodgkin lymphomas (MOAb for immunophenotyping of lymphocytes: CD14, CD45, CD3, CD19, CD4, CD8, CD16/56, HLA DR, CD38, CD25, CD23, CD5, CDl1c, CD30, light chain immunoglobulins). PMID- 9206930 TI - [Community-acquired pneumonia--current status of pathogen diagnosis]. AB - Procedures for the microbiological diagnosis of acute community-acquired pneumonia are based on the expected pathogens. Although a great variety of microorganisms are able to cause community-acquired pneumonia only a few pathogens play an important role in daily practice. The most important investigations are blood cultures and sputum cultures to detect bacteria like pneumococci, Haemophilus influenzae and Staphylococcus aureus as well as antibody tests for Mycoplasma pneumonia and Chlamydia pneumonia. According to anamnesis and clinic presentation tests such as for Legionella or viruses have to be added. Sometimes also rare pathogens have to be considered such as Coxiella burnetii, Leptospira, Hantaviruses, cryptococci or Chlamydia psittaci. The standard procedure for diagnosis of tuberculosis is the microscopical examination and the standardized culture in liquid and on solid media. Amplification methods such as PCR are also useful for a rapid diagnosis. However, the application of amplification procedures alone without culture is not recommended. PMID- 9206931 TI - American Gastroenterology Association issues guidelines for colorectal cancer screening. PMID- 9206932 TI - [Liver abscess caused by Streptococcus intermedius, following hemorrhoidectomy]. AB - The pyogenic liver abscess is an uncommon but potentially lethal complication of colo-anal surgery. The authors report a case due to Streptococcus intermedius, a pathogen with a known ability to produce visceral abscesses, after haemorrhoidectomy. According to the French consensus conference, the patient had received a prophylactic preoperative antibiotic regimen consisting of metronidazole, active against S intermedius. Despite surgical therapy and adequate antibiotics, the patient died of hepatic failure. PMID- 9206933 TI - [Idiosyncrasy of urology]. PMID- 9206934 TI - [Orthotopic bladder replacement: I. Physiopathology of orthotopic bladder replacement with intestine]. AB - OBJECTIVE: To analyze the physiopathological principles of utilizing the bowel for orthotopic bladder substitution and their effects on metabolism, function and continence. METHODS: The world literature is reviewed and our experience of 100 cases is described in the third part of this study. RESULTS/CONCLUSION: To reduce the metabolic changes, utilization of colonic or ileal segments with a maximum length of about 40 cm is advocated. This length of detubulized intestinal segment permits creating an ample, low pressure reservoir with an antireflux mechanism. The precise incidence of neoplastic degeneration of the ileal and colonic reservoirs is not known, but appears to be lower for the ileal neobladder. PMID- 9206935 TI - [Upper urinary tract and urethral tumors in patients with bladder carcinoma]. AB - OBJECTIVE: A clinicopathological study was conducted in patients with urothelial tumor of the bladder and urothelial neoplasm of the upper urinary tract or urethra to analyze the histological features, time interval between diagnoses and prognosis of these associated lesions. METHODS: 338 patients were reviewed retrospectively. A descriptive study was performed, including tables on the associated lesions and survival data. RESULTS: 21 cases (6.2%) had two urothelial tumors: carcinoma of the bladder associated with tumor of the upper urinary tract (14 cases) or urethra (7 cases). There is a longer time interval between diagnoses when the extravesical tumor is diagnosed after than when it is diagnosed before the bladder tumor (p < .05). A correlation between the grade, but not stage, of the extravescial and bladder tumors has been observed. We found no statistically significant differences for survival in patients with two tumors or with bladder cancer alone. Although not statistically significant, patients with metachronous extravescial tumor have a better prognosis than those with synchronous extravesical tumor. CONCLUSION: Patients with bladder cancer should be followed for years and screened for extravesical urothelial tumor. The foregoing associated lesion, especially the metachronous extravesical lesion, apparently does not make the prognosis worse. PMID- 9206936 TI - [Analysis of prognostic factors in superficial bladder tumors]. AB - OBJECTIVE: When the traditional prognostic factors (tumor grade, stage, size, number and cytological findings) are used as guidelines for intravesical therapy of superficial bladder tumors and the reported results are compared, it is not uncommon to find unexplainable differences. This study was conducted to determine the prognostic factors for tumor recurrence and progression before instituting any type of adjuvant therapy for superficial bladder tumors. METHODS: 81 consecutive patients with primary superficial bladder cancer (stage Ta-T1, grade 1-3) were entered into a surveillance protocol and controlled for a mean period of 14 months (range 3-44). Patient individual features (sex and age) and tumor characteristics (grade, stage, size, number, cytological findings) were analyzed to determine the risk of tumor recurrence and/or progression. RESULTS: Logistic regression analysis identified age as the only independent prognostic factor for recurrence, which was 6.37 fold (2-42) more frequent for subjects aged 65 years or older. Given the low progression rate (3 cases; 4.8%), a formal risk analysis could not be performed. CONCLUSION: The factors used to predict recurrence (tumor grade, stage, size, number and cytological findings) were not found to be independent in the present series. Certain predictors of tumor aggressiveness such as age-, which could modify tumor biology, were found to predispose to tumor recurrence. PMID- 9206937 TI - [Epididymal adenomatoid tumors. 2 new cases]. AB - OBJECTIVE: To review the different aspects of epididymal tumors, with special reference to adenomatoid tumor of the epididymis (mesothelial neoplasm). Two such cases are reported herein. METHODS: Two cases of adenomatoid tumor of the epididymis are described. The clinical features, diagnosis and treatment are discussed, within the context of the algorithm recently presented by Pellice and co-workers. RESULTS/CONCLUSION: Most of the epididymal tumors are benign and treatment using a scrotal approach will suffice. However, inguinotomy should be performed when the benign nature of the tumor cannot be established unequivocally. PMID- 9206939 TI - [The use of the EKL-COMPACT lithotriptor in the endoscopic treatment of ureteral lithiasis]. AB - OBJECTIVE: To report on our experience with the EKL-Compact (Olympus) lithotriptor in the treatment of ureteral calculi. METHODS/RESULTS: From December, 1994 to December, 1995, we performed 192 ureteroscopy procedures for endoscopic treatment of ureteral calculi. Stone removal was achieved without necessitating fragmentation in 131 cases. In the remaining cases, stone fragmentation with the EKL-Compact (Olympus) electrokinetic lithotriptor achieved a 95% success rate. CONCLUSION: We have achieved a high success rate with the EKL Compact lithotriptor. Its components are easily assembled, which facilitates sterilization and storage. It is compact and easily transportable. PMID- 9206938 TI - [Correlation between computed axial tomography and ileum obturating lymphadenectomy in localized adenocarcinoma of the prostate]. AB - OBJECTIVE: To correlate the findings of CT and ileoobturator lymphadenectomy in patients with localized adenocarcinoma of the prostate. METHODS: 94 patients with adenocarcinoma of the prostate were evaluated. Ileoobturator lymphadenectomy and brachytherapy were performed in 61.1%, radical prostatectomy in 22.5% and lymphadenectomy with prostatic labeling for subsequent external radiation therapy in 5%. Lymph node CT and pathology findings were correlated. RESULTS: Of 92 patients with a normal CT scan, 18 had positive nodes and 19.1% were understaged. Two patients with a CT scan suggestive of metastatic adenopathy had negative pathology findings. Seventy-two of the 92 patients with normal CT scans had negative nodes, accounting for a specificity of 76.6%. CONCLUSION: Pelvic lymph node involvement changes the prognosis of prostate cancer. However, the ability of CT to detect lymph node metastasis is limited. It is therefore not a reliable method and raises the costs of staging unnecessarily. PMID- 9206940 TI - [Urinary lithiasis in transplanted kidney]. AB - OBJECTIVE: We reviewed the records of patients submitted to renal transplantation at our institution to determine the incidence and risk factors for calculus formation in these patients. METHODS: Of 794 functioning renal grafts that had been transplanted from January, 1981 to May, 1996, 16 patients (2%), 9 males and 7 females, had upper urinary tract calculi post-transplantation. All 16 patients had received kidneys from cadaver donors. Three had donor graft lithiasis. The calculi were located predominantly in the calyces, at multiple sites in 7 patients and the mean size was 8.3 mm. The composition of the calculi was predominantly uric acid. Four patients who developed sudden obstructive anuria with elevated serum creatinine, underwent percutaneous drainage. RESULTS: All patients had one or more stone-predisposing factors, such as obstructive uropathy, recurrent urinary tract infection or metabolic abnormalities (predominantly hyperuricosuria). Five patients passed their stones spontaneously, 7 patients with uric acid stones were treated with urinary alkalinization, two patients underwent open pyelolithotomy, one underwent percutaneous nephrolithotomy and one patient with a small asymptomatic caliceal stone was managed conservatively (watchful waiting). During long-term follow-up (mean 69 months), 4 patients lost the real graft [only one case was related to urinary calculi (primary hyperoxaluria)] and 4 patients had recurrent calculi. CONCLUSION: Urinary lithiasis after renal transplantation is a relatively uncommon complication. A multifactorial etiology for calculus formation has been observed. The predisposing factors and composition of the calculi, but not frequency, are identical to those of non-transplant patients. A variety of methods are used to treat posttransplant calculi. The least invasive treatment available should be utilized according to the likelihood of recurrence and the need to preserve renal function. With adequate treatment and prophylaxis, posttransplant urolithiasis does not appear to affect graft function. PMID- 9206941 TI - [Role of percutaneous endopyelotomy in the treatment of pyeloureteral junction stenosis after failed ureteropyeloplasty]. AB - OBJECTIVE: To determine the value of percutaneous endopyelotomy (PE) in the treatment of ureteropelvic junction (UPJ) obstruction, particularly after failed ureteropelvioplasty. METHODS: A series of 14 (PE) procedures performed at our institution were retrospectively studied. The patients were divided into two groups: group I comprised patients in whom PE was used for primary repair and group II comprised patients submitted to PE after failed ureteropelvioplasty. The patients were assessed for hydronephrosis, lithiasis, previous procedures performed, operating time, duration of hospital stay and outcome. The Fisher test and Mann-Whitney U test were used for statistical analysis. RESULTS: The success of PE did not correlate with the degree of hydronephrosis, the presence of calculus or a previous procedure. The operating times were similar for both groups and for patients with and without lithiasis. The results were better for group II than for group I (p = 0.01). CONCLUSIONS: 1. In correctly selected cases, PE appears to be a good method of treatment for recurrent UPJ obstruction after failed ureteropelvioplasty. 2. The difficulty and the operating time of PE for recurrent UPJ obstruction is not different to that of previously untreated cases. No factor was found to correlate more with the success or failure of PE. PMID- 9206942 TI - [Radiopacity of double-J ureteral catheters. Computed tomography quantification and clinical implications]. AB - OBJECTIVE: To determine whether knowledge about the degree of radiopacity of the double-J ureteral catheters utilized in Spanish hospitals suffices for correct radiologic control and subsequent follow-up. METHODS: The CT attenuation index was utilized to determine the degree of radiopacity of 23 double-J ureteral stents, comprised of different biomaterials and from different manufacturers, that are used in Spanish hospitals. RESULTS: The values ranged from 1,000-3,070 Hounsfield units. CONCLUSION: All the catheters analyzed were sufficiently radiopaque to permit good radiologic control during insertion and subsequent follow-up. The radiopacity of the stent depends on the metal salt employed during the manufacturing process and not on the biomaterial. PMID- 9206943 TI - [Current status of the study of genital sympathetic evoked potentials in the assessment of impotence]. AB - OBJECTIVE: To determine the usefulness of penile sympathetic skin response (PSSR) in the study of impotence. METHODS: The PSSR, hand sympathetic skin response (HSSR), filling videocystography and SPACE (single potential analysis cavernous electromyography) were performed in 39 patients referred for study of impotence. RESULTS: A relationship between not obtaining PSSR and an open bladder neck (60%) in the videocystography at filling, and between a closed bladder neck and obtained PSSR (81%) was demonstrated. The percentage of normal hand sympathetic potentials was similar for patients with obtained and not obtained PSSR. The relationship between the degree of activity of the SPACE and the type of PSSR could not be demonstrated. CONCLUSION: The determination of the PSSR allows us to evaluate the sympathetic cavernous innervation. This would obviate performing a filling videocystography in the study of the neuroandrologic profile in impotence. The information obtained by PSSR is independent of that obtained from SPACE, therefore both procedures complement each other in the neuroandrologic study of impotence. PMID- 9206944 TI - [Assessment of sensory thresholds of the penile dorsal nerve as screening technique for neurologic lesion in impotence]. AB - OBJECTIVE: The present study was conducted to determine the usefulness of the perception and stimulation thresholds of the penile dorsal nerve in the diagnosis of neurogenic impotence. METHODS: A study was conducted to determine the neuroandrologic profile in 130 patients. According to the results of the neuroandrologic profile, they were classified as patients without neurogenic impotence (44 pts.; 34%) or with neurogenic impotence (86 pts.; 66%). The perception and stimulation thresholds (expressed in milliAmperes) were determined in all patients. Furthermore, the perineal sensation was clinically tested. Both thresholds were also studied in relation to a demonstrated neurologic lesion (sympathetic, parasympathetic, afferent pudendal, efferent pudendal and suprasacral lesions). RESULTS: Assessment of the perineal sensation demonstrated a high specificity and a low sensitivity in the diagnosis of neurogenic impotence. A significant difference was observed between both groups for the perception (confidence interval of difference between the neurogenic and non neurogenic group; from 0.02 to 4.95 mA) and stimulation thresholds (from 2.0 to 11 mA). Morever, the stimulation threshold was significantly higher in patients with alteration of the perineal sensation). Significant differences in the perception threshold were also demonstrated between patients with and without demonstration of efferent lesion, and in the stimulation threshold between patients with and without demonstration of pudendal afferent, pudendal efferent and sympathetic lesion. These differences are attributed to the presence of mixed lesions. However, given the width of the interval of normal values, it was not possible to find a useful cut-off point in both sensory thresholds. It could only be determined that the maximum value of the perception threshold in healthy subjects should be less than 9.45 mA. CONCLUSION: The sensorial thresholds of the electrical stimulation of the penile dorsal nerve and the data from the physical examination of perineal sensation are not useful for the diagnosis of neurogenic impotence. It is necessary to carry out a complete neuroandrologic profile. PMID- 9206945 TI - [A new solution to the retained Foley catheter]. AB - OBJECTIVE: To describe a new, effective and simple method to remove Foley catheters retained in the bladder. METHODS: After stretching the ureter, it is held close to the meatus with Kocher clamps. The catheter is cut transversely and disobstructed. The liquid utilized to inflate the balloon is removed and the catheter withdrawn. RESULTS: The method is easy, fast, indolent, low-cost, non traumatic, does not impede subsequent maneuvers and effective. CONCLUSION: This method is suitable after eliminating the valve mechanism and before recurring to balloon puncture or invasive methods. PMID- 9206946 TI - [Transformation of glandular cystitis into bladder transitional carcinoma with adenocarcinoma areas]. AB - OBJECTIVE: The association of cystitis glandularis with primary bladder tumors is well known. A case of cystitis glandularis progressing to transitional cell carcinoma is described and the literature briefly reviewed. METHODS/RESULTS: We report on a case of diffuse cystitis glandularis in whom progression to transitional cell carcinoma with areas of adenocarcinoma had been discovered. Treatment was by radical cystectomy with urinary diversion. CONCLUSION: Extensive and diffuse cystitis glandularis can be a potentially malignant lesion. Patients with diffuse cystitis glandularis should have a regular cystoscopic follow-up for early detection of progression to a bladder tumor. PMID- 9206947 TI - [Inverted carcinoma of kidney pelvis coexisting with transitional cell carcinoma of the bladder. A therapeutic alternative]. AB - OBJECTIVE: To describe a case of inverted carcinoma of the right renal pelvis associated with transitional cell carcinoma of the bladder. The unusual nature of this condition and the incidence of tumor recurrence in some patients with bladder cancer and, at least initially, no histopathological evidence of infiltrating tumor prompted us to report this case. METHODS/RESULTS: A 65-year old male consulted for prostatism. Several months earlier he had had an episode of hematuria. The difficulty in making the diagnosis is underscored and the possibility of conservative surgical management is discussed. Tumor recurrence at two sites distinct from that of the bladder tumor warranted TUR. Two years after the first surgical procedure the patient is disease-free. CONCLUSION: Like most authors, we usually perform radical surgery in these cases. However, in the case described herein, the difficulty in making the diagnosis prompted us to use a conservative approach which, at the present time, appears to have been effective for the renal tumor. New treatment options, like endourological surgery, may have an important place in the management of this tumor type in carefully selected cases. PMID- 9206948 TI - [Sertoli cell tumor of the testis. Report of a case]. AB - OBJECTIVE: To describe an additional case of Sertoli cell testicular tumor. Although more cases have been reported in the world literature, only one case with bilateral involvement has been described in the Spanish literature over the last 18 years. METHODS: A 34-year-old male consulted for a testicular mass which he had detected a few weeks earlier. US and CT evaluation disclosed a tumor in the right testis and no evidence of metastasis. The patient underwent right radical orchidectomy via the inguinal approach and insertion of a testicular prosthesis. RESULTS: Histopathological analysis of the surgical specimen disclosed a Sertoli cell testicular tumor. Patient control follow-up at 3 and 9 months postoperatively showed no changes in tumor markers or CT findings. CONCLUSION: The benign or malignant nature of Sertoli cell testicular tumor is currently defined by the presence or absence of metastasis, although there are histopathological criteria that might indicate a certain degree of aggressiveness. PMID- 9206949 TI - [Burned out testicular tumor]. AB - OBJECTIVE: Extragonadal germ cell tumor is a rare condition. The present study reports an additional case of this uncommon disease. METHODS/RESULTS: A 24-year old male with retroperitoneal disease and a testicular lesion detected by ultrasonography is described. Histopathological analysis of the orchiectomy specimen disclosed a 'burned out' tumor. The anatomopathological features of this tumor type are discussed. CONCLUSION: Extragonadal germ cell tumor is a rare condition. It derives from the germ cells that exist in the gonad and is located in every anatomical region but the gonad itself. Ultrasonography must be performed if an extragonadal germ cell tumor is suspected. PMID- 9206950 TI - [Bilateral isolated infiltration in the epididymis and sperm cord as first sign of chronic lymphatic leukemia]. AB - OBJECTIVE: Involvement of extralymphatic tissue in chronic lymphocytic leukemia (CLL) is uncommon and that of the epididymis and spermatic cord is rare. This paper describes an unusual case with metastasis to both epididymis and the spermatic cord as the first sign of reactivation of CLL. METHODS/RESULTS: A patient with CLL is described. The first sign of disease progression was the involvement of both epididymis and the spermatic cord. The literature on tumors of the epididymis and spermatic cord and CLL is briefly reviewed. CONCLUSION: Metastatic tumors of the epididymis and spermatic cord are rare. The importance of the pathological findings in making the diagnosis is underscored. PMID- 9206951 TI - [Retroperitoneal ganglioneuroma. Report of a case]. AB - OBJECTIVE: To report an additional case of retroperitoneal ganglioneuroma. METHODS/RESULTS: A case of retroperitoneal ganglioneuroma that had been incidentally discovered in a 27-year-old male during abdominal US evaluation is described. The diagnosis was based on the histopathological findings after US guided biopsy. The clinical features and the findings of the complementary tests, which included radiological assessment, intravenous urography, CT and cavography are presented, as well as the pathology findings. Treatment was by complete surgical excision of the tumor. CONCLUSION: Treatment of ganglioneuroma is by surgery, since the diagnosis is generally based on the histopathological analysis of the surgical specimen. For those cases with a preoperative diagnosis, some authors advocate surgery for patients with clinical evidence of neuroblastoma or another pathology arising from this disease. Although infrequent, ganglioneuroma coexisting with neuroblastoma can occur. For this reason, complete excision of the tumor is preferred. PMID- 9206952 TI - [Diphtheritic polyneuropathy: clinico-morphologic study]. AB - 18 patients with grave DP treated with a long-term artificial pulmonary ventilation and feeding through a naso-gastric probe were studied. Biopsies of n. suralis taken at different periods after the appearance of the first signs of DP (from the 19th to the 69th day) were studied at light and electronic microscopy. The basis of DP is toxic myelopathy with paranodal demyelination mainly in the large myelinated neural fibers and a segmentary one in the smaller neural fibers. Axonal degeneration was observed in the gravest cases of DP and was secondary being the result of axon squeezing by a folded myelin and voluminous Schwann cell cytoplasm invaginated into the axon. In no case of DP inflammatory changes and(or) involvement of the immunocompetent cells were found. There was pronounced proliferation and activation of Schwann cells due to intensive utilization of the degradation products of myelin and remyelinization. Morphological signs of remyelinization were observed on the 35th day of DP in the presence of enhancing neurological symptoms. But remyelinization was not complete even on the 69th day of DP. PMID- 9206953 TI - [Electron microscopic and immunomorphologic study of the placenta in genital mycoplasmosis]. AB - Peculiar ultrastructural features allowed the authors to reveal M. Homminis corpuscles in placental tissues of women with genital mycoplasmosis. Mycoplasma was found in the amniotic epithelium, chorionic plate, in the lumen of villous capillaries, this showing possible hematogenic way of the infection from mother to fetus. In the placenta of women with genital mycoplasmosis immunopathological processes develop in association with formation of pathogenic immune complexes (PIC) that are fixing the C3 complement fraction. Location of PIC on the syncytiotrophoblast membranes and vascular endothelium causes immunological inflammation with the involvement of immunocompetent cells resulting in the destruction of syncytial membranes and membranes of the placental barrier. Damage to the placental barrier membranes promotes the development of placental failure, a complicated course of pregnancy and delivery, deterioration of the state of the fetus and newborn infant. PMID- 9206955 TI - [Clinico-morphologic characterization of mucoepidermoid carcinoma of the salivary glands]. AB - 64 cases of SGMC were studied. Clinical morphology of epidermoid carcinoma was studied according to the 2nd edition of the International histologic classification of the tumors of this site (WHO, Geneva, 1990). This allows to study not only incidence of this tumor depending on the site, sex and age, but to give new data on its biology. The malignancy of all the three types of this tumor (of low, moderate and high grade) was confirmed by means of histologic, histochemical, electron-microscopic (EM) and EM-histochemical methods. The previous assumption on benign character of the low-grade variant of the tumor was not confirmed. PMID- 9206954 TI - [Immunomorphologic characterization of various parameters of stomach cancer invasion]. AB - 25 cases of advanced stomach carcinoma were studied immunomorphologically with the use of the antibodies panel to the carcinoembryonic antigen and meconian antigen B1, collagen types IV, I and III, laminin, Ki-67, P-105, factor VIII. Secretory and proliferative activity of tumor cells was shown unrelated to histological structure and degree of tumor differentiation. The more was proliferative activity the weaker was the secretory function. Formation of the basal membrane (BM), the degree of collagen formation and angiogenesis in the tubular adenocarcinoma did not depend on the differentiation level and the degree of tumor cells secretory activity. On the contrary, carcinoid component was characterized by pronounced angiogenesis and tendency to correlation between the degree of differentiation and the degree of BM formation. Stomach carcinoma is a heterogeneous group of tumors whose various morphological features may have an independent prognostic value. PMID- 9206956 TI - [Mediator aspects of the inflammatory process]. AB - Signal sequences of inflammation and regeneration are presented. The number of factors such as sensor neuropeptides, lipid mediators, interleukins and factors of growth are shown to participate in the initiation and regulation of inflammation. These factors are produced in a certain sequence by various structures: cells-object of damage, opsonocytophagic and immune cells, connective tissue cells. This results in the stimulation of adhesion proteins, processes of migration, chemotaxis, interaction with components of the intercellular matrix and parenchyma cells of the viscera. Fibroblast is considered to be the central figure in the resolution of inflammation and its chronization. The regulation of the process occurs with the help of autocrine regulation and apoptosis of functionally exhausted cells. PMID- 9206957 TI - [Age-related changes in the autonomic ganglia]. AB - Ultrastructure of vegetative ganglions (neck-thoracic, intracardial and intestinal) of Wistar and SHR rats, 26-28 months of age was studied electron microscopically. The most pronounced changes were found in the neck-thoracic ganglions where, apart from lipofuscin deposits, lamellar bodies were frequently found. Redistribution of neuromediators in the neuron body and an increase of the neuroactive substances release into the intercellular space of the ganglion occur with age. The data on possible postsynaptic influence on the presynapse by means of neuromediators are presented. Nissl bodies hypertrophy was observed in the neck-thoracic and intracardial ganglions of old SHR rats. PMID- 9206958 TI - [Morphometric and ultrastructural characterization of the bronchial smooth musculature in experimental bronchospasm]. AB - A new technique was used in experimental bronchospasm model on 30 guinea pigs: spot alkaline dissociation of bronchial smooth myocytes. No pronounced smooth muscle hypertrophy occurred within 50 days but the number of myocytes with contractures increased as well as proliferative activity and hyperplasia of myocytes, degeneration of large myocytes and restructuring of myocyte population. PMID- 9206959 TI - [Morphometric characterization of intestinal metaplasia of the mucosa of the pyloric antrum section of the stomach in ulcer disease]. AB - 71 stomachs removed because of ulcer were examined. It was confirmed that atrophic changes and intestinal metaplasia of the stomach mucous membrane are more pronounced in the stomach ulcer than in duodenal ulcer. There were no significant quantitative differences in the state of glandular apparatus in the anterior wall and small curvature of the stomach. The mean content of the border enterocytes in stomach ulcer was 78.8%, duodenal ulcer-86.9%: goblet cells-15.3% and 9.9%, Panet cells-2.7 and 1.14%, respectively, enterochromaffin cells-3.2 and 2.06%, respectively. PMID- 9206960 TI - [Congenital developmental defects negatively affecting the viability of children of Kursk region: prevalence, pattern, trends]. AB - The distribution of and time trends in congenital defects of development (CD) influencing viability of children in the Kursk region were studied on the basis of 4517 autopsy reports for the period 1985-1994. A total of 880 children had CD (5.40%). The incidence of CD in towns (6.38%), was higher than in country regions (4.91%), p < 0.001, being the highest in the central parts of the region. Among the stillborns the specific proportion of congenital defects was 14.53%, perinatal deaths occurred in 16.96%, deaths during the 1st year of life in 22.10%. The percentage of multiple congenital defects, the defects of cardiovascular system, developmental anomalies of digestive system, the defects of central nervous system and organs of sense was 39.77, 27.16, 10.11 and 8.30, respectively. PMID- 9206961 TI - [Morphologic diagnosis of various forms of thymus gland diseases in children using morphometry]. AB - Quantitative criteria of obscure thymic pathology (OTP) are obtained by morphometric study of the thymus of 102 infants with severe infectious inflammatory processes (IIP). OTP is characterised by depression of epithelial hyperplasia in accidental involution found in 28.4% IIP and is followed by the disturbances mainly in the cell immunity system. This type of thymus pathology is associated in 69% of cases with congenital anomaly of the viscera and skeleton this allowing to regard it as a form of thymus fetal dysplasia. PMID- 9206962 TI - [Breast cancer with neuroendocrine differentiation]. AB - One rare case of mammary carcinoma with an unusual clinical course and non typical morphology is described. The diagnosis of mammary carcinoma with neuro endocrine differentiation was established on the basis of histologic, immunomorphologic and electron-microscopic examination. PMID- 9206963 TI - [Microscopic identification of melanoma metastases in occult primary tumor]. AB - A case of the metastasis of the tumor with a high degree of anaplasia to the neck lymph nodes is described. Histological features of the neoplasm--a combination of the epithelial-like and fusiform cells, alveolar and individual character of their surrounding by the argyrophil fibers and the presence of fine argyrophil granules in Paskal silvering--allowed to suspect the metastasis of pigment-free malignant melanoma, and to select a small panel of antibodies for immunohistochemical analysis. The expression by the tumor cells of vimentin, S 100 protein and melanoma antigen proved this tumor to be a malignant melanoma metastasis. Location of the primary tumor has not been revealed. PMID- 9206965 TI - [A case of Schilder's disease]. AB - Schilder's disease is diffuse periaxial encephalitis, a rare condition from the group of demyelinating diseases. The characteristic features are: demyelinization of the white matter, lymphocytic perivascular infiltrates, microglial proliferation with final fibrillar gliosis. Progressive disturbances of the nervous activity, paralysis, convulsion and exitus letalis are observed clinically. One case of this disease in a female of 40 is described which was thought to be schizophrenia and was not diagnosed clinically. PMID- 9206964 TI - [Mesenchymal tumor of the small pelvis with hypoglycemic syndrome]. AB - In spite of a large size, mesenchymal tumor of the small pelvis was not diagnosed clinically, and hypoglycemia was considered as a manifestation of hyperinsulinism. The tumor was identified as mesenchymal, most likely fibrosarcoma, immunohistochemically. The use of antibodies against insulin, glucagon and somatostatin showed the disturbance in the balance of cells in the hyperplastic islands of the pancreas. Multifocal hyperplasia of the islands is considered as a compensatory adaptive reaction to paraneoplastic extrapancreatic hypoglycemia. PMID- 9206966 TI - [Acceleration of histologic tissue processing and decalcification using a microwave oven]. AB - Feasibility of significant acceleration of histoprocessing by using domestic microwave oven is shown. Two histoprocessing protocols for a single bit and many bits of surgery, biopsy and autopsy materials have been introduced. The design of a special container is described. The histoprocessing period was reduced to 2 hrs. Significant acceleration (10 to 20 times) of decalcification of bone tissue by using domestic microwave oven was achieved. Decalcification took from 3 to 8 hrs in good quality of the microscopic images. PMID- 9206967 TI - [Medical insurance classification of iatrogenic diseases and various approaches to its evaluation]. PMID- 9206968 TI - [The problem of regeneration of the gastrointestinal tract mucosa in erosive ulcerous lesions]. AB - Disturbance of regeneration in the above pathological condition results from the damage to tunica propria and loss of its function for the support of structural homeostasis. Disintegration of the basal membrane and extracellular matrix (main components in the epithelial stromal interactions) is of a primary importance under these conditions. PMID- 9206969 TI - [Dendritic antigen-presenting cells of the oral mucosa in health and in pathological conditions]. AB - Oral mucosa contains a system of dendritic antigen-presenting cells which provide its specific immune defence. These cells are found both in mucosal epithelium and lamina propria and possess specific ultrastructural, cytochemical, immunocytochemical and functional properties. Langerhans cells are the most numerous and well-studied population of antigen-presenting cells in oral mucosa. However, epithelium and lamina propria harbour other types of dendritic cells. Dendritic cells trap antigens process them and provide them to lymphocytes; they also influence other mucosal cell types. This review summarizes recent data on distribution, morphologic and functional characteristics of dendritic antigen presenting cells of the oral mucosa in health and in some pathological conditions. A possible role of these cells in pathogenesis of some diseases is discussed. PMID- 9206970 TI - [General pathology of trace element deficiency]. AB - According to the authors' concept, different forms of trace element deficiency show some general rules of development. All of them are followed by a decrease of immune resistance. Trace element deficiency is never isolated, it is always characterized by trace element unbalance and is followed by a considerable disturbance of metabolism (mineral, lipid, carbohydrate and protein) with relevant manifestations. Reduced immune resistance and pluriglandular endocrinopathy create the conditions for various malignancies. PMID- 9206971 TI - Vascular reactivity following ischemia/reperfusion. AB - Data in the literature supports the hypothesis that reactive oxygen species generated in the vascular wall alter vascular regulation. At present the majority of the literature tends to suggest that oxidant induced damage on the smooth muscle cell impair vasoconstriction. However, direct action of oxidants on the smooth muscle cell impair vasoconstrictor function. Differences in studies in the literature are likely to be reconciled when the target sites of reactive oxygen species are considered. Future research in this area should lead to a more comprehensive understanding on the impact of these pathways on vasoregulation in postischemic tissue. PMID- 9206972 TI - Immune response of neonates elicited by somatic transgene vaccination with naked DNA. AB - Neonates display lower immune responsiveness and higher susceptibility for high dose tolerance. Quantitative as well as functional differences between the neonatal and adult lymphocytes or antigen presenting cells (APC) respectively, explain the particular immune responsiveness during the early stages of the postnatal development. Reduced numbers of lymphocytes and APCs as well as a modified responsiveness of T cells in neonates, are the main factors that account for the low threshold of tolerance in newborns. Taking into account these particularities, the design of effective vaccines for neonates poses significant difficulties. We hypothesized that a continuous exposure to low doses of antigens may avoid high-zone tolerance and may lead instead, to effective expansion of effector and memory cells. Indeed, inoculation of newborn mice with plasmids encoding nucleoprotein (NP) or hemagglutinin (HA) of influenza virus, led to the priming of specific cytotoxic (CTL), helper (Th) and B cells, rather than induction of unresponsiveness. Mice immunized as neonates with naked DNA and challenged later with lethal doses of influenza virus, displayed significant protection. Thus, DNA immunization may be a promising strategy for vaccination against serious infectious diseases of infants and children. PMID- 9206973 TI - Molecular mechanism of actin-dependent retrograde flow in lamellipodia of motile cells. AB - In motile, eukaryotic cells, a variety of cell-associated material (collectively termed here as 'particles') continuously flows, relative to the substratum, from the front to the back of the extreme margin of the cell (termed the 'lamellipodium'). This retrograde particle flow, occurs both over the surface of, and inside the lamellipodium. Force to drive retrograde particle flow in lamellipodia is dependent on actin filaments, but the precise mechanism of force generation, and function of the flow is generally not well understood. Actin filaments themselves, in lamellipodia of most motile cell types studied also flow retrograde relative to the substratum. This actin flow, in Aplysia bag cell neuronal growth cones, is known to be driven by activity of a myosin. In these growth cones, retrograde flow of cell surface-attached particles is coupled to retrograde actin flow. In Aplysia, force from retrograde actin flow may limit certain types of growth cone motility. In other motile cell types, such as keratocytes and fibroblasts, the mechanism of retrograde particle flow and function of retrograde actin flow in lamellipodia is poorly understood. For these cell types, recent data provide a basis for proposing alternative actin-based mechanisms to drive retrograde particle flow in lamellipodia. One mechanism is based on activity of a putative pointed end- directed actin motor, and the other on tension-driven surface lipid flow. Here I will review recent advances that have been made in determining the molecular mechanism of force generation to drive retrograde particle flow relative to the substratum in lamellipodia of motile cells. I will address the function of retrograde actin flow in lamellipodia, and apparent differences between Aplysia and other motile cell types. PMID- 9206975 TI - Reamplification of differential display products: more is not better. AB - Differential display polymerase chain reaction is technically challenging at a number of steps, including reliable reamplification of differentially expressed sequences after they are isolated from an acrylamide gel. As the only source of the sequence found to be of interest is the gel from which it was identified as being differentially expressed, failed attempts at reamplification can be particularly frustrating. In our laboratory, attempts to reamplify DNA sequences cut from a denaturing 6% acrylamide gel consistently failed when greater than 5 microliters of eluted product was used as template in a subsequent PCR reaction. If less template was used in subsequent PCR reactions, reamplification was consistently successful. This observation emphasized the importance of using limited amounts of template when reamplifying sequences that are differentially displayed on denaturing acrylamide gels PMID- 9206974 TI - Genes implicated in the pathogenesis of Alzheimer's disease. AB - Both the early and late-onset Alzheimer's disease affect millions of people throughout the world. A number of molecules have been implicated in the pathogenesis of Alzheimer's disease. These include presenilin 1 and 2 (PS1 and PS2), a beta-amyloid peptide, and tau protein. Presenilin 1 and 2 genes implicated in the early-onset familial Alzheimer's disease have been cloned. Both PS1 and PS2 are integral membrane proteins and may function as receptors or channel proteins. Missense mutations in PS1 and PS2 genes have been found in families that cosegregate with early-onset Alzheimer's disease. Overexpression of the mutated PS1 gene produced amyloid plaques in the brain of transgenic mice. Secreted beta-amyloid protein similar to that in the senile plaques of Alzheimer's disease was found to be elevated in fibroblast media from subjects with PS1 or PS2 mutations. Transgenic mice which carried the mutant form of the beta-amyloid precursor protein gene expressed high concentrations of mutant copy of the gene and exhibited abundant amyloid plaques in the brain and memory loss. The mutated PS2 gene enhanced apoptotic activity. This enhanced apoptotic activity may accelerate the process of neurodegeneration leading to an earlier age in the onset of the disease. Identification of lesions in the molecules that are important in the Alzheimer's disease should allow developing therapeutic approaches for its treatment. PMID- 9206978 TI - An improved method for Southwestern blotting. AB - We have developed a modified Southwestern blotting technique which utilizes broad spectrum protease inhibitors during nuclear protein extraction and a procedure for radiolabelling an oligonucleotide probe to a high specific activity. These modifications have resulted in minimal protein degradation during nuclear protein isolation and have permitted room temperature hybridizations, improving both the facility and sensitivity of the standard Southwestern assay. This technique was used in our laboratory to visualize NFAT-1 consensus sequence binding proteins in nuclear extracts of human promyelocytic HL-60 cells. PMID- 9206979 TI - Inhibition of the HIV Rev transactivator : a new target for therapeutic intervention. AB - The viral transactivator Rev is essential for HIV replication, since it allows the nuclear export of unspliced and partially spliced viral mRNAs that encode the structural proteins. Rev is an RNA binding protein that interacts with a highly structured RNA element, the RRE, found within the envelope sequences. This viral protein also interacts with cellular proteins, termed nucleoporins, and acts as an adaptor between the viral mRNAs and the cellular nuclear export machinery. Both interactions are specific, and required for Rev function. Because of its crucial role in the HIV replication cycle, and its novel mechanism of action, Rev represents an ideal target for therapeutic intervention. This review describes the efforts towards Rev inhibition. Gene therapy approaches, including the expression of trans-dominant mutants and RNA decoys, as well as antisense therapies and small molecule inhibitors of Rev-RRE binding or Rev interaction with the cellular machinery will be discussed PMID- 9206980 TI - Fatty acid transporters in animal cells. AB - The mechanism by which fatty acids transverse the plasma membrane has been a controversial subject. Kinetic studies of fatty acid uptake suggested the presence of a protein carrier system in certain cells which exhibit rapid fatty acid influx and/or efflux such as hepatocytes, adipocytes and jejunal mucosal cells. Five plasma membrane proteins have been identified and proposed as candidates for fatty acid transporters thus far. These includes: Plasma Membrane Fatty Acid Binding Protein (FABPpm), Fatty Acid Translocase (FAT), caveolin, a 56 kDa renal fatty acid binding protein and Fatty Acid Transport Protein (FATP). The first four proteins were identified by classical biochemical techniques while FATP, the one most recently reported, was identified by expression cloning strategies. Each of these proteins has distinct primary amino acid sequence and tissue-specific pattern of expression. It remains to be determined whether the proteins identified to date function as individual polypeptides or as a single component of a larger complex. This review summarizes recent advances concerning the structure, function and regulation of these putative fatty acid transporters. PMID- 9206981 TI - Interactions between superoxide and nitric oxide: implications in DNA damage and mutagenesis. AB - Chronic inflammation is known to be associated with enhanced production of both nitric oxide (NO) and reactive oxygen species such as superoxide (O2-) and hydrogen peroxide (H2O2). Patients with long-standing ulcerative colitis are also known to be at increased risk of developing colorectal cancer. Although NO and reactive oxygen intermediates alone have been known to damage DNA and to promote a wide array of mutagenic reactions, there is increasing evidence to suggest that the interaction between O2- and NO may dictate the type of mutagenic reactions produced at sites where both these free radicals are produced. In the absence of O2-, NO will engage in nitrosative chemistry to yield stable N-nitrosamine derivatives of secondary amines and promote nitrosative deamination of DNA bases. As the flux of O2- is increased, nitrosation reactions are suppressed and oxidative chemistry is enhanced. Thus, depending upon the fluxes of each radical either nitrosation or oxidation chemistry may predominate. The fundamental understanding between O2- and NO may provide new insight in the mechanisms responsible for inflammation-induced mutagenesis. PMID- 9206982 TI - Recent advances in lymphocyte signaling and regulation. AB - The antigen receptor signaling pathway in lymphocytes is vital to their development and biological function. Recent studies have shown that protein tyrosine kinases and phosphatases are essential components in this receptor signaling pathway and therefore, are critical for the development of mature and functionally competent lymphocytes. The Src kinase family of protein tyrosine kinases coordinates the early signaling events in antigen receptor signaling via phosphorylation of tyrosine-based substrates. These kinases are regulated by the concerted action of the Csk family of non-receptor protein tyrosine kinases and the protein tyrosine phosphatase, CD45. A complex set of phosphorylation and dephosphorylation events regulate protein tyrosine kinase activity. Upon antigen stimulation, Src protein tyrosine kinases in conjunction with the tyrosine kinases, ZAP-70 and Syk initiate downstream effectors leading to Ca2+ mobilization, the activation of the Ras pathway and transcriptional activation. The roles of the various adapter proteins in these pathways are now being elucidated. It is apparent that a network of phosphorylation events connect the antigen receptor to intracellular signaling pathways. PMID- 9206984 TI - Neurochemical basis of disruption of hippocampal long term potentiation by chronic alcohol exposure. AB - The aim of this review is to summarize the possible mechanisms underlying the long-term impairment of learning and memory resulting from chronic ethanol treatment (CET) especially that involving decrements in long-term potentiation (LTP) in hippocampus. CET for a 28-week duration affects the rat hippocampal formation in such a way as to decrease the magnitude of LTP; an effect that can last as long as 7 months after ethanol withdrawal. It appears that NMDA receptor number in hippocampus is unchanged after CET whereas the data suggest a more pronounced role for changes in GABAergic and cholinergic synaptic transmission in determining how CET influences the induction of LTP in hippocampus. In particular, changes in presynaptic modulation of neurotransmitter release in hippocampus may be one mechanism by which CET inhibits LTP. Thus, the mechanisms underlying the effect of CET on LTP are a result of changes in a number of neurotransmitter systems in hippocampus (GABAergic and cholinergic) rather than based solely on changes in glutamate transmission. PMID- 9206985 TI - The role of Langerhans islets in pancreatic ductal adenocarcinoma. AB - An intimate relationship between the exocrine and endocrine pancreas has been convincingly demonstrated in recent years. Animal experiments have shed some light into the complex dialog between the two tissues. This interaction is pronounced in diseases of the pancreas, especially in experimentally-induced and human pancreatic cancers. New evidence highlights the importance of intact islets in the development of exocrine pancreatic cancer. Although tumors arise from large and small ducts, invasive and malignant adenocarcinomas of ductal phenotype also derive from stem cells within islets. Development of cancer within islets explains the association between pancreatic cancer and impaired glucose tolerance or diabetes. Hence, the previous epidemiological studies suggesting that diabetes is a predisposing factor for pancreatic cancer are refuted. The available evidence suggests that pancreatic cancer in a large number of pancreatic cancer patients ultimately leads to diabetes, and that removal of the tumors improves or cures the diabetes. Both in the hamster pancreatic cancer model and in patients, the development of cancer is associated with elevated plasma levels of islet amyloid polypeptide, which may be used as a tumor marker. PMID- 9206986 TI - Weekly paclitaxel as a radiation sensitizer for locally advanced gastric and pancreatic cancers: the Brown University Oncology Group experience. AB - Many patients with cancer of the stomach or pancreas have locally advanced, unresectable disease at diagnosis or will develop an early local or regional recurrence despite potentially curative surgery. Effective local treatment could increase the proportion of patients able to undergo surgery and decrease locoregional recurrences, which should improve overall survival. External beam radiation (RT) by itself has little effect. Standard treatment, such as RT with concurrent administration of 5-fluorouracil-based chemotherapy as a radiation sensitizer, has, at best, a modest impact on locoregional recurrences and survival. The use of a more effective radiosensitizer might improve the efficacy of local treatment. Paclitaxel synchronizes cells at G2M, the phase of the cell cycle during which cells are most sensitive to the effects of ionizing radiation, and has been demonstrated to sensitize a variety of human cell lines to the effects of RT. In patients with locally advanced non-small cell lung cancer (NSCLC), the Brown University Oncology Group (BrUOG) has demonstrated a high response rate to low-dose weekly paclitaxel with concurrent RT. In addition, we demonstrated that the response to paclitaxel/RT was not affected by mutations in the p53 tumor suppressor gene. This suggested that paclitaxel/RT would be a rational treatment approach for other malignancies with a high frequency of p53 mutations, such as gastric and pancreatic cancers. We have completed a phase I study of weekly paclitaxel and concurrent radiation for locally advanced gastric and pancreatic cancers. The maximum tolerated dose of paclitaxel was 50mg/m2/week for six weeks with 50 Gray (Gy) abdominal radiation. The dose limiting toxicities were abdominal pain, nausea and anorexia. Preliminary response data from ongoing phase II studies suggest that preoperative paclitaxel/RT has substantial activity in patients with locally advanced gastric and pancreatic cancers, though whether this will translate into improved disease-free and overall survival in these patients is not known. PMID- 9206987 TI - T cell signaling of macrophage function in inflammatory disease. AB - Macrophages play diverse roles in episodic T cell-mediated inflammatory diseases such as multiple sclerosis and rheumatoid arthritis, function as accessory cells for T cell activation, as pro-inflammatory cells, as effector cells which mediate tissue damage, and as anti-inflammatory cells which promote wound healing. In addition to the many roles of T cell-derived cytokines in differentially modulating these diverse macrophage activities, research over the last few years has demonstrated that contact-dependent signaling which occurs during T cell macrophage adhesion is a critical triggering event in the activation of macrophage function. Substantial research emphasis has been placed on CD40 as a mediator of contact dependent signaling. However, other membrane-anchored receptor:ligand pairs may also contribute to the stimulation of macrophage function. This is a brief review of the rapidly expanding, but still incomplete, knowledge of how T cells, through both contact-dependent and cytokine signals, regulate macrophage function during inflammatory disease. PMID- 9206988 TI - Transgenic rabbit models for the study of atherosclerosis. AB - The rabbit rapidly develops severe hypercholesterolemia leading to premature atherosclerosis in response to dietary manipulation. Transgenic rabbit models with altered expression of specific genes will provide new approaches to understanding the underlying mechanisms that contribute to this disease. Recently established lines of transgenic rabbits that overexpress hepatic lipase and apolipoprotein E are yielding fresh insights into the functions of these proteins and their role in lesion development PMID- 9206990 TI - The carboxy-terminal domain of human surfactant protein B is not required for secretion in milk of transgenic mice. AB - Previous studies in which human pulmonary surfactant protein B (SP-B) was targeted to the mammary gland of transgenic mice using the rat whey acidic protein (WAP) regulatory sequences resulted in secretion of only the unprocessed proprotein (42 kDa) in milk. To test the feasibility of producing a partially processed SP-B protein in milk, a new construct was designed in which the coding region for the carboxy-terminal domain was deleted. Expression of rWAP/SP-BDELTA C mRNA was detected in all three transgenic lines generated, and the expected carboxy-terminal deleted SP-B molecule (28 kDa), identified by using domain specific antibodies, was secreted in the milk. Histochemical examination of lactating mammary tissue from the transgenic line expressing the highest levels of WAP/SP-BDelta C mRNA revealed an inhibition of lobulo-alveolar development, and led to growth retardation in pups, apparently due to the decreased milk production. Mothers from this line tended to cannibalize litters in mid lactation. This phenotype has been observed previously with several other WAP based transgenes. This phenotype suggests that there may be an upper limit to the level of SP-BDeltAC which can be produced in milk PMID- 9206991 TI - Elevation of extracellular magnesium rapidly raises intracellular free Mg2+ in human aortic endothelial cells: is extracellular Mg2+ a regulatory cation? AB - Extracellular magnesium ions [Mg2+]o are known to regulate functions of endothelial cells, but whether [Mg2+]o can alter intracellular free ionized magnesium [Mg2+]i in these cells remains unknown. The present studies, using digital imaging microscopy and the Mg2+ fluorescent probe, mag-fura-2, determined effects of elevation of [Mg2+]o on [Mg2+]i in cultured human aortic endothelial cells. With normal Mg2+(1.2 mM)-containing incubation media, [Mg2+]i was 0.51+/ 0.04 mM with a heterogeneous distribution. The ratio of [Mg2+]i/[Mg2+]o was 0.52+/-0.07. Elevation of [Mg2+]o up to 4.8 mM increased [Mg2+]i to 0.80+/-0.07 mM in 2-10 min and lowered the ratio of [Mg2+]i/[Mg2+]o to 0.16+/-0.02. Irrespective of the observed increments of [Mg2+]i, a subcellular heterogeneous distribution of [Mg2+]i was always evident in all cells tested. Our results suggest that [Mg2+]o can regulate [Mg2+]i more rapidly than heretofore believed, supporting the hypothesis that extracellular Mg2+ can exert regulatory effects on endothelial cell functions and probably act as extracellular regulatory cations PMID- 9206992 TI - Evasion of cytotoxic T lymphocyte (CTL) responses by nef-dependent induction of Fas ligand (CD95L) expression on simian immunodeficiency virus-infected cells. AB - Inoculation of macaques with live attenuated SIV strains has been shown to protect against subsequent challenge with wild-type SIV. The protective mechanism(s) remain obscure. To study the effect in more detail, we have investigated the role of virus-specific CTL responses in macaques infected with an attenuated SIV strain (pC8), which has a four-amino acid deletion in the nef gene, as compared with the wild-type SIVmac32H clone (pJ5). Cynomolgus macaques infected with pC8 were protected against subsequent challenge with pJ5 and did not develop any AIDS-like symptoms in the 12 months after infection. The pC8 induced protection was associated with high levels of virus-specific CTL responses to a variety of viral antigens. In contrast, pJ5-infected macaques had little, if any, detectable CTL response to the viral proteins after three months. The latter group of macaques also showed increased Fas expression and apoptotic cell death in both the CD4(+) and CD8(+) populations. In vitro, pJ5 but not pC8 leads to an increase in FasL expression on infected cells. Thus the expression of FasL may protect infected cells from CTL attack, killing viral-specific CTLs in the process, and providing a route for escaping the immune response, leading to the increased pathogenicity of pJ5. pC8, on the other hand does not induce FasL expression, allowing the development of a protective CTL response. Furthermore, interruption of the Fas-FasL interaction allows the regeneration of viral specific CTL responses in pJ5-infected animals. This observation suggests an additional therapeutic approach to the treatment of AIDS. PMID- 9206993 TI - Commitment of immature CD4+8+ thymocytes to the CD4 lineage requires CD3 signaling but does not require expression of clonotypic T cell receptor (TCR) chains. AB - As a consequence of positive selection in the thymus, immature CD4(+)8(+) double positive, [DP] thymocytes selectively terminate synthesis of one coreceptor molecule and, as a result, differentiate into either CD4(+) or CD8(+) T cells. The decision by individual DP thymocytes to terminate synthesis of one or the other coreceptor molecule is referred to as lineage commitment. Previously, we reported that the intrathymic signals that induced commitment to the CD4 versus CD8 T cell lineages were markedly asymmetric. Notably, CD8 commitment appeared to require lineage-specific signals, whereas CD4 commitment appeared to occur in the absence of lineage-specific signals by default. Consequently, it was unclear whether CD4 commitment, as revealed by selective termination of CD8 coreceptor synthesis, occurred in all DP thymocytes, or whether CD4 commitment occurred only in T cell receptor (TCR)-CD3-signaled DP thymocytes. Here, we report that selective termination of CD8 coreceptor synthesis does not occur in DP thymocytes spontaneously. Rather, CD4 commitment in DP thymocytes requires signals transduced by either CD3 or zeta chains, which can signal CD4 commitment even in the absence of clonotypic TCR chains. PMID- 9206994 TI - The central executioner of apoptosis: multiple connections between protease activation and mitochondria in Fas/APO-1/CD95- and ceramide-induced apoptosis. AB - According to current understanding, cytoplasmic events including activation of protease cascades and mitochondrial permeability transition (PT) participate in the control of nuclear apoptosis. However, the relationship between protease activation and PT has remained elusive. When apoptosis is induced by cross linking of the Fas/APO-1/CD95 receptor, activation of interleukin-1beta converting enzyme (ICE; caspase 1) or ICE-like enzymes precedes the disruption of the mitochondrial inner transmembrane potential (DeltaPsim). In contrast, cytosolic CPP32/ Yama/Apopain/caspase 3 activation, plasma membrane phosphatidyl serine exposure, and nuclear apoptosis only occur in cells in which the DeltaPsim is fully disrupted. Transfection with the cowpox protease inhibitor crmA or culture in the presence of the synthetic ICE-specific inhibitor Ac-YVAD.cmk both prevent the DeltaPsim collapse and subsequent apoptosis. Cytosols from anti-Fas treated human lymphoma cells accumulate an activity that induces PT in isolated mitochondria in vitro and that is neutralized by crmA or Ac-YVAD.cmk. Recombinant purified ICE suffices to cause isolated mitochondria to undergo PT-like large amplitude swelling and to disrupt their DeltaPsim. In addition, ICE-treated mitochondria release an apoptosis-inducing factor (AIF) that induces apoptotic changes (chromatin condensation and oligonucleosomal DNA fragmentation) in isolated nuclei in vitro. AIF is a protease (or protease activator) that can be inhibited by the broad spectrum apoptosis inhibitor Z-VAD.fmk and that causes the proteolytical activation of CPP32. Although Bcl-2 is a highly efficient inhibitor of mitochondrial alterations (large amplitude swelling + DeltaPsim collapse + release of AIF) induced by prooxidants or cytosols from ceramide-treated cells, it has no effect on the ICE-induced mitochondrial PT and AIF release. These data connect a protease activation pathway with the mitochondrial phase of apoptosis regulation. In addition, they provide a plausible explanation of why Bcl-2 fails to interfere with Fas-triggered apoptosis in most cell types, yet prevents ceramide- and prooxidant-induced apoptosis. PMID- 9206995 TI - Interleukin 12 (IL-12) is crucial to the development of protective immunity in mice intravenously infected with mycobacterium tuberculosis. AB - Immunity to Mycobacterium tuberculosis infection is associated with the emergence of protective CD4 T cells that secrete cytokines, resulting in activation of macrophages and the recruitment of monocytes to initiate granuloma formation. The cytokine-mediating macrophage activation is interferon-gamma (IFN-gamma), which is largely dependent on interleukin-12 (IL-12) for its induction. To address the role of IL-12 in immunity to tuberculosis, IL-12 p40(-/-) mice were infected with M. tuberculosis and their capacity to control bacterial growth and other characteristics of their immune response were determined. The IL-12 p40(-/-) mice were unable to control bacterial growth and this appeared to be linked to the absence of both innate and acquired sources of IFN-gamma. T cell activation as measured by delayed type hypersensitivity and lymphocyte accumulation at the site of infection were both markedly reduced in the IL-12 p40(-/-) mice. Therefore, IL 12 is essential to the generation of a protective immune response to M. tuberculosis, with its main functions being the induction of the expression of IFN-gamma and the activation of antigen-specific lymphocytes capable of creating a protective granuloma. PMID- 9206996 TI - 4-1BB costimulatory signals preferentially induce CD8+ T cell proliferation and lead to the amplification in vivo of cytotoxic T cell responses. AB - The 4-1BB receptor is an inducible type I membrane protein and member of the tumor necrosis factor receptor (TNFR) superfamily that is rapidly expressed on the surface of CD4+ and CD8+ T cells after antigen- or mitogen-induced activation. Cross-linking of 4-1BB and the T cell receptor (TCR) on activated T cells has been shown to deliver a costimulatory signal to T cells. Here, we expand upon previously published studies by demonstrating that CD8+ T cells when compared with CD4+ T cells are preferentially responsive to both early activation events and proliferative signals provided via the TCR and 4-1BB. In comparison, CD28-mediated costimulatory signals appear to function in a reciprocal manner to those induced through 4-1BB costimulation. In vivo examination of the effects of anti-4-1BB monoclonal antibodies (mAbs) on antigen-induced T cell activation have shown that the administration of epitope-specific anti-4-1BB mAbs amplified the generation of H-2d-specific cytotoxic T cells in a murine model of acute graft versus host disease (GVHD) and enhanced the rapidity of cardiac allograft or skin transplant rejection in mice. Cytokine analysis of in vitro activated CD4+ and CD8+ T cells revealed that anti-4-1BB costimulation markedly enhanced interferon gamma production by CD8+ T cells and that anti-4-1BB mediated proliferation of CD8+ T cells appears to be IL-2 independent. The results of these studies suggest that regulatory signals delivered by the 4-1BB receptor play an important role in the regulation of cytotoxic T cells in cellular immune responses to antigen. PMID- 9206997 TI - Resistance to fas-mediated apoptosis of peripheral T cells in human T lymphocyte virus type I (HTLV-I) transgenic mice with autoimmune arthropathy. AB - Transgenic mice carrying the env-pX region of human T lymphocyte virus type I (HTLV-I) develop autoimmune arthropathy in high incidence. Adopting the approach that Fas-mediated apoptosis has a critical function in the elimination of self reactive T cells, we examined the involvement of this apoptosis in the induction of autoimmunity in HTLV-I transgenic mice. Splenic T cells derived from the transgenic mice were more resistant to apoptosis induced by anti-Fas mAb than those of the nontransgenic mice, whereas no appreciable difference in apoptosis was detected for thymocytes from either mouse's type. The resistance of transgenic T cells may be due to Tax coded in the pX region, since Tax mediates the inhibition of anti-Fas- induced apoptosis in mature T cell line, Jurkat. Among the transgenic mice, the extent of the resistance to Fas-mediated apoptosis was further enhanced in transgenic T cells with disease. These results suggest that the escape of self-reactive T cells from Fas-mediated apoptosis in the periphery, is critical for the development of autoimmune arthropathy in HTLV-I transgenic mice. PMID- 9206998 TI - Induction of a CD8+ cytotoxic T lymphocyte response by cross-priming requires cognate CD4+ T cell help. AB - Class I-restricted presentation is usually associated with cytoplasmic degradation of cellular proteins and is often considered inaccessible to exogenous antigens. Nonetheless, certain exogenous elements can gain entry into this so-called endogenous pathway by a mechanism termed cross-presentation. This is known to be effective for class I-restricted cytotoxic T lymphocyte (CTL) cross-priming directed against a variety of exogenous tumor, viral, and minor transplantation antigens. The related effect of cross-tolerance can also effectively eliminate responses to selected self components. In both cases, this presentation appears to require the active involvement of a bone marrow-derived antigen presenting cell (APC). Here, we show that CTL induction by cross-priming with cell-associated ovalbumin requires the active involvement of CD4+ helper T cells. Importantly, this CD4+ population is only effective when both the helper and CTL determinants are recognized on the same APC. Moreover, we would argue that the cognitive nature of this event suggests that the CD4+ T cell actively modifies the APC, converting it into an effective stimulator for the successful priming of the CTL precursor. PMID- 9206999 TI - Transferable anergy: superantigen treatment induces CD4+ T cell tolerance that is reversible and requires CD4-CD8- cells and interferon gamma. AB - Bacterial superantigens induce peripheral unresponsiveness in CD4+ T cell populations that express appropriate Vbeta chains. We have used Vbeta3/Valpha11 T cell receptor transgenic (Tg) mice and the Vbeta3-specific superantigen staphylococcal enterotoxin A (SEA) to further investigate the mechanisms that contribute to such unresponsiveness. As in other models, in vivo exposure to SEA rendered the Tg CD4+ cells unresponsive to subsequent restimulation in vitro with antigen or mitogens. However, when the SEA-treated CD4+ cells were completely purified away from all other contaminating cells, they regained the ability to proliferate and secrete cytokines. Moreover, enriched CD4-CD8- cells from the SEA treated mice suppressed the responses of fresh control CD4+ cells in mixed cultures indicating that the apparent "anergy" was both transferable and reversible. Further analysis demonstrated that interferon gamma, but not the Fas receptor, played a critical role in the suppression. PMID- 9207000 TI - Epitope-dependent selection of highly restricted or diverse T cell receptor repertoires in response to persistent infection by Epstein-Barr virus. AB - The T cell receptor (TCR) repertoires of cytotoxic responses to the immunodominant and subdominant HLA A11-restricted epitopes in the Epstein-Barr virus (EBV) nuclear antigen-4 were investigated in four healthy virus carriers. The response to the subdominant epitope (EBNA4 399-408, designated AVF) was highly restricted with conserved Vbeta usage and identical length and amino acid motifs in the third complementarity-determining regions (CDR3), while a broad repertoire using different combinations of TCR-alpha/beta V and J segments and CDR3 regions was selected by the immunodominant epitope (EBNA4 416-424, designated IVT). Distinct patterns of interaction with the A11-peptide complex were revealed for each AVF- or IVT-specific TCR clonotype by alanine scanning mutagenesis analysis. Blocking of cytotoxic function by antibodies specific for the CD8 coreceptor indicated that, while AVF-specific TCRs are of high affinity, the oligoclonal response to the IVT epitope includes both low- and high-affinity TCRs. Thus, comparison of the memory response to two epitopes derived from the same viral antigen and presented through the same MHC class I allele suggests that immunodominance may correlate with the capacity to maintain a broad TCR repertoire. PMID- 9207001 TI - A role for CD4 in peripheral T cell differentiation. AB - Naive CD4+ T helper cells (Th) differentiate into one of two well-defined cell types during immune responses. Mature Th1 and Th2 cells regulate the type of response as a consequence of the unique cytokines that they secrete. CD4 serves a prominent role in potentiating antigen recognition by helper T cells. We have examined the role of CD4 in peripheral T cell differentiation by studying helper T cells from mice with a congenital defect in CD4 expression. After protein immunization or infection with Leishmania major, CD4-deficient mice were incapable of mounting antigen-specific Th2 responses, but retained their Th1 potency. CD4-deficient, T cell receptor transgenic T cells were also incapable of Th2 differentiation after in vitro activation. Expression of a wild-type CD4 transgene corrected the Th2 defect of CD4-deficient mice in all immune responses tested. To investigate the role of the cytoplasmic domain, mice reconstituted with a truncated CD4 molecule were also studied. Expression of the tailless CD4 transgene could not rescue the Th2 defect of CD4-deficient mice immunized with protein or CD4-deficient transgenic T cells activated in vitro, raising the possibility that the cytoplasmic domain of CD4 may influence Th2 generation. Expression of the tailless transgene was, however, capable of restoring Th2 development in CD4-deficient mice infected with L. major or CD4-deficient transgenic T cells activated in the presence of recombinant IL-4, demonstrating that the cytoplasmic domain is not absolutely required for Th2 development. Together, these results demonstrate a previously undescribed role of the CD4 molecule. The requirement for CD4 in Th2 maturation reflects the importance of molecules other than cytokines in the control of helper T cell differentiation. PMID- 9207003 TI - An interleukin 5 mutant distinguishes between two functional responses in human eosinophils. AB - Interleukin 5 (IL-5) is the key cytokine involved in regulating the production and many of the specialized functions of mature eosinophils including priming, adhesion, and survival. We have generated a point mutant of human IL-5, IL-5 (E12K), which is devoid of agonist activity in both a TF-1 cell proliferation assay and a human eosinophil adhesion assay. However, IL-5 (E12K) is a potent and specific antagonist of both these IL-5-dependent functional responses. In both receptor binding and cross-linking studies the wild-type and IL-5 (E12K) mutant exhibit virtually identical properties. This mutant protein was unable to stimulate tyrosine phosphorylation in human eosinophils, and blocked the phosphorylation stimulated by IL-5. In contrast, IL-5 (E12K) is a full agonist in a human eosinophil survival assay, although with reduced potency compared to the wild-type protein. This IL-5 mutant enables us to clearly distinguish between two IL-5-dependent functional responses and reveals distinct mechanisms of receptor/cellular activation. PMID- 9207002 TI - Requirements for CD1d recognition by human invariant Valpha24+ CD4-CD8- T cells. AB - A subset of human CD4-CD8- T cells that expresses an invariant Valpha24-JalphaQ T cell receptor (TCR)-alpha chain, paired predominantly with Vbeta11, has been identified. A series of these Valpha24 Vbeta11 clones were shown to have TCR-beta CDR3 diversity and express the natural killer (NK) locus-encoded C-type lectins NKR-P1A, CD94, and CD69. However, in contrast to NK cells, they did not express killer inhibitory receptors, CD16, CD56, or CD57. All invariant Valpha24(+) clones recognized the MHC class I-like CD16 molecule and discriminated between CD1d and other closely related human CD1 proteins, indicating that recognition was TCR-mediated. Recognition was not dependent upon an endosomal targeting motif in the cytoplasmic tail of CD1d. Upon activation by anti-CD3 or CD1d, the clones produced both Th1 and Th2 cytokines. These results demonstrate that human invariant Valpha24+ CD4-CD8- T cells, and presumably the homologous murine NK1+ T cell population, are CD1d reactive and functionally distinct from NK cells. The conservation of this cell population and of the CD1d ligand across species indicates an important immunological function. PMID- 9207004 TI - Polarization of chemokine receptors to the leading edge during lymphocyte chemotaxis. AB - Leukocyte migration in response to cell attractant gradients or chemotaxis is a key phenomenon both in cell movement and in the inflammatory response. Chemokines are quite likely to be the key molecules directing migration of leukocytes that involve cell polarization with generation of specialized cell compartments. The precise mechanism of leukocyte chemoattraction is not known, however. In this study, we demonstrate that the CC chemokine receptors CCR2 and CCR5, but not cytokine receptors such as interleukin (IL)-2Ralpha, IL-2Rbeta, tumor necrosis factor receptor 1, or transforming growth factor betaR, are redistributed to a pole in T cells that are migrating in response to chemokines. Immunofluorescence and confocal microscopy studies show that the chemokine receptors concentrate at the leading edge of the cell on the flattened cell-substratum contact area, induced specifically by the signals that trigger cell polarization. The redistribution of chemokine receptors is blocked by pertussis toxin and is dependent on cell adhesion through integrin receptors, which mediate cell migration. Chemokine receptor expression on the leading edge of migrating polarized lymphocytes appears to act as a sensor mechanism for the directed migration of leukocytes through a chemoattractant gradient. PMID- 9207005 TI - Identification of CCR8: a human monocyte and thymus receptor for the CC chemokine I-309. AB - The human CC chemokine I-309 is a potent monocyte chemoattractant and inhibits apoptosis in thymic cell lines. Here, we identify a specific human I-309 receptor, and name it CCR8 according to an accepted nomenclature system. The receptor has seven predicted transmembrane domains, is expressed constitutively in monocytes and thymus, and is encoded by a previously reported gene of previously unknown function named, alternatively, CY6, TER1, and CKR-L1. After transfection with the CY6 open reading frame, a mouse pre-B cell line exhibited calcium flux and chemotaxis in response to I-309 (EC50 = 2 nM for each), whereas 20 other chemokines were inactive. Signaling was sensitive to pertussis toxin, suggesting coupling to a Gi-type G protein. These properties parallel those of endogenous I-309 receptors expressed in an HL-60 clone 15 cell line model. The apparent monogamous relationship between I-309 and CCR8 is unusual among known CC chemokines and known CC chemokine receptors. CCR8 may regulate monocyte chemotaxis and thymic cell line apoptosis. PMID- 9207006 TI - Upregulation of Fas ligand by simian immunodeficiency virus - a nef-arious mechanism of immune evasion? PMID- 9207007 TI - An antagonist of monocyte chemoattractant protein 1 (MCP-1) inhibits arthritis in the MRL-lpr mouse model. AB - An antagonist of human monocyte chemoattractant protein (MCP)-1, which consists of MCP-1(9-76), had previously been characterized and shown to inhibit MCP-1 activity in vitro. To test the hypothesis that, by inhibiting endogenous MCP-1, the antagonist has antiinflammatory activity in vivo, we examined its effect in the MRL-lpr mouse model of arthritis. This strain spontaneously develops a chronic inflammatory arthritis that is similar to human rheumatoid arthritis. Daily injection of the antagonist, MCP-1(9-76), prevented the onset of arthritis as monitored by measuring joint swelling and by histopathological evaluation of the joints. In contrast, controls treated with native MCP-1 had enhanced arthritis symptoms, indicating that the inhibitory effect is specific to the antagonist. In experiments where the antagonist was given only after the disease had already developed, there was a marked reduction in symptoms and histopathology, although individuals varied in the magnitude of the response. The mechanism of inhibition of disease is not known, although the results suggest that it could be more complex than the competitive inhibition of ligand binding that is observed in vitro. The demonstration of the beneficial effects of an MCP 1 antagonist in arthritis suggests that chemokine receptor antagonists could have therapeutic application in inflammatory diseases. PMID- 9207008 TI - HIV coreceptor downregulation as antiviral principle: SDF-1alpha-dependent internalization of the chemokine receptor CXCR4 contributes to inhibition of HIV replication. AB - Ligation of CCR5 by the CC chemokines RANTES, MIP-1alpha or MIP-1beta, and of CXCR4 by the CXC chemokine SDF-1alpha, profoundly inhibits the replication of HIV strains that use these coreceptors for entry into CD4(+) T lymphocytes. The mechanism of entry inhibition is not known. We found a rapid and extensive downregulation of CXCR4 by SDF-1alpha and of CCR5 by RANTES or the antagonist RANTES(9-68). Confocal laser scanning microscopy showed that CCR5 and CXCR4, after binding to their ligands, are internalized into vesicles that qualify as early endosomes as indicated by colocalization with transferrin receptors. Internalization was not affected by treatment with Bordetella pertussis toxin, showing that it is independent of signaling via Gi-proteins. Removal of SDF 1alpha led to rapid, but incomplete surface reexpression of CXCR4, a process that was not inhibited by cycloheximide, suggesting that the coreceptor is recycling from the internalization pool. Deletion of the COOH-terminal, cytoplasmic domain of CXCR4 did not affect HIV entry, but prevented SDF-1alpha-induced receptor downregulation and decreased the potency of SDF-1alpha as inhibitor of HIV replication. Our results indicate that the ability of the coreceptor to internalize is not required for HIV entry, but contributes to the HIV suppressive effect of CXC and CC chemokines. PMID- 9207009 TI - Development of inflammatory angiogenesis by local stimulation of Fas in vivo. AB - Fas-Fas ligand interaction is thought to be a crucial mechanism in controlling lymphocyte expansion by inducing lymphocyte apoptosis. However, Fas is also broadly expressed on nonlymphoid cells, where its function in vivo remains to be determined. In this study, we describe the development of inflammatory angiogenesis induced by agonistic anti-Fas mAb Jo2 in a murine model where Matrigel is used as a vehicle for the delivery of mediators. The subcutaneous implants in mice of Matrigel containing mAb Jo2 became rapidly infiltrated by endothelial cells and by scattered monocytes and macrophages. After formation and canalization of new vessels, marked intravascular accumulation and extravasation of neutrophils were observed. Several mast cells were also detected in the inflammatory infiltrate. The phenomenon was dose and time dependent and required the presence of heparin. The dependency on activation of Fas is suggested by the observation that the inflammatory angiogenesis was restricted to the agonistic anti-Fas mAb and it was absent in lpr Fas-mutant mice. Apoptotic cells were not detectable at any time inside the implant or in the surrounding tissue, suggesting that angiogenesis and cell infiltration did not result from recruitment of phagocytes by apoptotic cells but rather by a stimulatory signal through Fas-engagement. These findings suggest a role for Fas-Fas ligand interaction in promoting local angiogenesis and inflammation. PMID- 9207010 TI - Treatment of experimental autoimmune encephalomyelitis with genetically modified memory T cells. AB - The migratory properties of memory T cells provide a model vector system for site specific delivery of therapeutic transgene factors to autoimmune inflammatory lesions. Lymph node cells from (SWRxSJL)F1 mice immunized with the p139-151 determinant of myelin proteolipid protein (PLP) were transfected with a DNA construct that placed the anti-inflammatory cytokine interleukin-10 (IL-10) cDNA under control of an antigen-inducible IL-2 promoter region. Isolated T cell clones demonstrated antigen-inducible expression of transgene IL-10 and expressed cell surface markers consistent with the phenotype of normal memory T cells. Upon adoptive transfer, transfected T cell clones were able to inhibit onset of experimental autoimmune encephalomyelitis (EAE) and to treat EAE animals therapeutically after onset of neurologic signs. Semiquantitative immunocytochemistry showed a significant correlation between decreased demyelination and treatment with the transfected T cells. Taken together, these data indicate the autoreactive T cells can be genetically designed to produce therapeutic factors in an antigen-inducible manner resulting in a decreased severity of clinical and histological autoimmune demyelinating disease. PMID- 9207011 TI - Identification of a DNA segment exhibiting rearrangement modifying effects upon transgenic delta-deleting elements. AB - Control of the rearrangement and expression of the T cell receptor alpha and delta chains is critical for determining T cell type. The process of delta deletion is a candidate mechanism for maintaining separation of the alpha and delta loci. Mice harboring a transgenic reporter delta deletion construct show alpha/beta T cell lineage-specific use of the transgenic elements. A 48-basepair segment of DNA, termed HPS1A, when deleted from this reporter construct, loses tight lineage-specific rearrangement control of transgenic elements, with abundant rearrangements of transgenic delta-deleting elements now in gamma/delta T cells. Furthermore, HPS1A augments recombination frequency of extrachromosomal substrates in an in vitro recombination assay. DNA binding proteins recognizing HPS1A have been identified and are restricted to early B and T cells, during the time of active rearrangement of endogenous TCR and immunoglobulin loci. These data are consistent with delta deletion playing an important role in maintaining separate TCR alpha and delta loci. PMID- 9207012 TI - Plant pathogenic RNAs and RNA catalysis. AB - The rolling circle replication of small circular plant pathogenic RNAs requires a processing step to convert multimeric intermediates to monomers which are then circularized. Eleven such RNAs are known so far, two are viroids, one is viroid like and the remainder are satellite RNAs dependent on a helper virus for replication. The processing step is RNA-catalysed in all cases, at least in vitro. All plus forms of these RNAs self-cleave via the hammerhead structure whereas only eight of the minus RNAs self-cleave, five via the hammerhead structure and three via the hairpin structure. There are about 20 other viroids where the processing mechanism has yet to be determined but they are likely candidates for a new type of self-cleavage reaction which is predicted to be conserved in all these viroids. Hepatitis delta RNA is the only circular pathogenic RNA known to self-cleave in the animal kingdom. It is feasible that more single-stranded circular pathogenic RNAs are waiting to be discovered and these could be prospective for new types of self-cleavage reactions. PMID- 9207013 TI - Peptide nucleic acid (PNA) is capable of enhancing hammerhead ribozyme activity with long but not with short RNA substrates. AB - Long RNA substrates are inefficiently cleaved by hammerhead ribozymes in trans. Oligonucleotide facilitators capable of affecting the ribozyme activity by interacting with the substrates at the termini of the ribozyme provide a possibility to improve ribozyme mediated cleavage of long RNA substrates. We have examined the effect of PNA as facilitator in vitro in order to test if even artificial compounds have facilitating potential. Effects of 12mer PNA- (peptide nucleic acid), RNA- and DNA-facilitators of identical sequence were measured with three substrates containing either 942, 452 or 39 nucleotides. The PNA facilitator enhances the ribozyme activity with both, the 942mer and the 452mer substrate to a slightly smaller extent than RNA and DNA facilitators. This effect was observed up to PNA facilitator:substrate ratios of 200:1. The enhancement becomes smaller as the PNA facilitator:substrate ratio exceeds 200:1. With the 39mer substrate, the PNA facilitator decreases the ribozyme activity by more than 100-fold, even at PNA facilitator:substrate ratios of 1:1. Although with long substrates the effect of the PNA facilitator is slightly smaller than the effect of identical RNA or DNA facilitators, PNA may be a more practical choice for potential applications in vivo because PNA is much more resistant to degradation by cellular enzymes. PMID- 9207014 TI - Pleiotropic effects of intron removal on base modification pattern of yeast tRNAPhe: an in vitro study. AB - Cell-free yeast extract has been successfully used to catalyze the enzymatic formation of 11 out of the 14 naturally occurring modified nucleotides in yeast tRNAPhe(anticodon GAA). They are m2G10, D17, m22G26, Cm32, Gm34,psi39, m5C40, m7G46, m5C49, T54 andpsi55. Only D16, Y37 and m1A58 were not formed under in vitro conditions. However, m1G37was quantitatively produced instead of Y37. The naturally occurring intron was absolutely required for m5C40formation while it hindered completely the enzymatic formation of Cm32, Gm34and m1G37. Enzymatic formation of m22G26,psi39, m7G46, m5C49, T54 andpsi55were not or only slightly affected by the presence of the intron. These results allow us to classify the different tRNA modification enzymes into three groups: intron insensitive, intron dependent, and those requiring the absence of the intron. The fact that truncated tRNAPheconsisting of the anticodon stem and loop prolonged with the 19 nucleotide long intron is a substrate for tRNA: cytosine-40 methylase demonstrates that the enzyme is not only strictly intron dependent, but also does not require fully structured tRNA. PMID- 9207015 TI - Structure of pvu II DNA-(cytosine N4) methyltransferase, an example of domain permutation and protein fold assignment. AB - We have determined the structure of Pvu II methyltransferase (M. Pvu II) complexed with S -adenosyl-L-methionine (AdoMet) by multiwavelength anomalous diffraction, using a crystal of the selenomethionine-substituted protein. M. Pvu II catalyzes transfer of the methyl group from AdoMet to the exocyclic amino (N4) nitrogen of the central cytosine in its recognition sequence 5'-CAGCTG-3'. The protein is dominated by an open alpha/beta-sheet structure with a prominent V shaped cleft: AdoMet and catalytic amino acids are located at the bottom of this cleft. The size and the basic nature of the cleft are consistent with duplex DNA binding. The target (methylatable) cytosine, if flipped out of the double helical DNA as seen for DNA methyltransferases that generate 5-methylcytosine, would fit into the concave active site next to the AdoMet. This M. Pvu IIalpha/beta-sheet structure is very similar to those of M. Hha I (a cytosine C5 methyltransferase) and M. Taq I (an adenine N6 methyltransferase), consistent with a model predicting that DNA methyltransferases share a common structural fold while having the major functional regions permuted into three distinct linear orders. The main feature of the common fold is a seven-stranded beta-sheet (6 7 5 4 1 2 3) formed by five parallel beta-strands and an antiparallel beta-hairpin. The beta-sheet is flanked by six parallel alpha-helices, three on each side. The AdoMet binding site is located at the C-terminal ends of strands beta1 and beta2 and the active site is at the C-terminal ends of strands beta4 and beta5 and the N-terminal end of strand beta7. The AdoMet-protein interactions are almost identical among M. Pvu II, M. Hha I and M. Taq I, as well as in an RNA methyltransferase and at least one small molecule methyltransferase. The structural similarity among the active sites of M. Pvu II, M. Taq I and M. Hha I reveals that catalytic amino acids essential for cytosine N4 and adenine N6 methylation coincide spatially with those for cytosine C5 methylation, suggesting a mechanism for amino methylation. PMID- 9207016 TI - DNA gyrase can cleave short DNA fragments in the presence of quinolone drugs. AB - We have analysed the DNA cleavage reaction of DNA gyrase using oligonucleotides annealed to a single-stranded M13 derivative containing a preferred gyrase cleavage site. We find that gyrase can cleave duplexes down to approximately 20 bp in size in the presence of the quinolone drugs ciprofloxacin and oxolinic acid. Ciprofloxacin shows a variation in its site specificity with an apparent preference for G bases adjacent to the cleavage sites, whereas oxolinic acid stimulates cleavage predominantly at the previously determined site. With either drug, cleavage will not occur within 6 bases from the end of a DNA duplex or a nick. We suggest that cleavage site specificity with short DNA duplexes is determined by drug-DNA interactions whereas with longer fragments the positioning effect of the DNA wrap around gyrase prescribes the site of cleavage. PMID- 9207017 TI - Tetracycline-controlled transcription in eukaryotes: novel transactivators with graded transactivation potential. AB - Several tetracycline-controlled transactivators (tTA) were generated which differ in their activation potential by >3 orders of magnitude. The transactivators are fusions between the Tet repressor and minimal transcriptional activation domains derived from Herpes simplex virus protein 16 (VP16). By reducing the VP16 moiety of the previously described tTA to 12 amino acids, potential targets for interactions with various cellular transcription factors were eliminated, as were potential epitopes which may elicit a cellular immune response. When compared with the originally described tTA, these new transactivators are tolerated at higher intracellular concentrations. This will facilitate establishment of tet regulatory systems under a variety of conditions, but particularly when cell type restricted tetracycline-controlled gene expression is to be achieved in transgenic organisms via homologous recombination. PMID- 9207018 TI - A new peptide vector for efficient delivery of oligonucleotides into mammalian cells. AB - The development of antisense and gene therapy has focused mainly on improving methods for oligonucleotide and gene delivery into cells. In the present work, we describe a potent new strategy for oligonucleotide delivery based on the use of a short peptide vector, termed MPG (27 residues), which contains a hydrophobic domain derived from the fusion sequence of HIV gp41 and a hydrophilic domain derived from the nuclear localization sequence of SV40 T-antigen. The formation of peptide vector/oligonucleotide complexes was investigated by measuring changes in intrinsic tryptophan fluorescence of peptide and of mansyl-labelled oligonucleotides. MPG exhibits relatively high affinity for both single- and double-stranded DNA in a nanomolar range. Based on both intrinsic and extrinsic fluorescence titrations, it appears that the main binding between MPG and oligonucleotides occurs through electrostatic interactions, which involve the basic-residues of the peptide vector. Further peptide/peptide interactions also occur, leading to a higher MPG/oligonucleotide ratio (in the region of 20/1), which suggests that oligonucleotides are most likely coated with several molecules of MPG. Premixed complexes of peptide vector with single or double stranded oligonucleotides are delivered into cultured mammalian cells in less than 1 h with relatively high efficiency (90%). This new strategy of oligonucleotide delivery into cultured cells based on a peptide vector offers several advantages compared to other commonly used approaches of delivery including efficiency, stability and absence of cytotoxicity. The interaction with MPG strongly increases both the stability of the oligonucleotide to nuclease and crossing of the plasma membrane. The mechanism of cell delivery of oligonucleotides by MPG does not follow the endosomal pathway, which explains the rapid and efficient delivery of oligonucleotides in the nucleus. As such, we propose this peptide vector as a powerful tool for potential development in gene and antisense therapy. PMID- 9207019 TI - A biologically active 53 kDa fragment of overproduced alanyl-tRNA synthetase from Thermus thermophilus HB8 specifically interacts with tRNA Ala acceptor helix. AB - The alaS gene encoding the alanyl-tRNA synthetase (AlaRS) from Thermus thermophilus HB8 was cloned and sequenced. The gene comprises 2646 bp, corresponding to 882 amino acids, 45% of which are identical to the enzyme from Escherichia coli . The T. thermophilus AlaRS was overproduced in E.coli , purified and characterized. It has high thermal stability up to approximately 65 degrees C, with a temperature optimum of aminoacylation activity at approximately 60 degrees C, and will be valuable for crystallization. The purified enzyme appears as a dimer with a specific activity of 220 U/mg and k cat/ K M values of 118 000/s/M for alanine and 114 000/s/M for ATP. By genetic engineering a 53 kDa fragment of AlaRS comprising the N-terminal 470 amino acids (AlaN470) was also overproduced and purified. It is as stable as entire AlaRS and sufficient for specific aminoacylation of intact tRNAAla, as well as acceptor stem microhelices with a G3-U70, but not U3-A70, I3-U70 or C3-U70, base pair. The reduced binding strength of such microhelices to AlaN470 enabled, due to the resulting fast exchange of the microhelices between free and complexed states, preliminary NMR analyses of the binding mode and intermolecular recognition. PMID- 9207020 TI - PolyPhred: automating the detection and genotyping of single nucleotide substitutions using fluorescence-based resequencing. AB - Fluorescence-based sequencing is playing an increasingly important role in efforts to identify DNA polymorphisms and mutations of biological and medical interest. The application of this technology in generating the reference sequence of simple and complex genomes is also driving the development of new computer programs to automate base calling (Phred), sequence assembly (Phrap) and sequence assembly editing (Consed) in high throughput settings. In this report we describe a new computer program known as PolyPhred that automatically detects the presence of heterozygous single nucleotide substitutions by fluorescencebased sequencing of PCR products. Its operations are integrated with the use of the Phred, Phrap and Consed programs and together these tools generate a high throughput system for detecting DNA polymorphisms and mutations by large scale fluorescence-based resequencing. Analysis of sequences containing known DNA variants demonstrates that the accuracy of PolyPhred with single pass data is >99% when the sequences are generated with fluorescent dye-labeled primers and approximately 90% for those prepared with dye-labeled terminators. PMID- 9207021 TI - The leptin receptor promoter controls expression of a second distinct protein. AB - The leptin receptor (OB-R) is a single membrane- spanning protein that mediates the weight-regulatory effects of leptin (OB protein). Several mRNA splice variants have been described which either encode OB-R proteins with cytoplasmic domains of different length or the OB-R and B219/OBR variants, which have different 5'-untranslated regions. Here we report evidence for the synthesis of a human mRNA splice variant of the OB-R gene that potentially encodes a novel protein, leptin receptor gene-related protein (OB-RGRP), which displays no sequence similarity to the leptin receptor itself. This OB-RGRP transcript contains the first two OB-R gene 5'-untranslated exons, but then is alternatively spliced to two novel exons which were mapped to a yeast artificial chromosome containing the leptin receptor gene. First identified by analysis of a large human expressed sequence tag database, the OB-RGRP transcript has now also been found in human and mouse tissues by the use of PCR. Preliminary experiments suggest that OB-RGRP and the OB-R variants share similar patterns of expression that are distinct from that of the B219/OBR variant. OB-RGRP is highly homologous to putative open reading frames in both yeast and Caenorhabditis elegans , suggesting a phylogenetically conserved role for this novel protein. PMID- 9207022 TI - E2F-dependent mitogenic stimulation of the splicing of transcripts from an S phase-regulated gene. AB - There is one class of genes whose expression increases at the G1/S transition of the cell cycle. One of these genes codes for 6-phosphofructo-2-kinase/fructose 2,6-bisphosphatase (PFK-2), an enzyme that controls glycolysis. The cell-cycle regulation of the PFK-2 gene depends on a binding site for the transcription factor E2F located at the 5'end of the first exon and involves not only a transcriptional, but also a post-transcriptional, mechanism. We have investigated this mechanism by studying in Rat-1 fibroblasts mature and immature mRNAs from the endogenous PFK-2 gene and from stably expressed transgenes containing PFK-2 gene regions. An increase in precursor mRNA half-life and processing took place at the G1/S transition. Transgenes with a mutated E2F binding site or with mutated splice sites lost the regulation by serum, indicating that both an intact E2F binding site and an efficient splicing reaction are necessary for proper mitogenic stimulation. In quiescent cells, the transgene lacking the E2F binding site was more efficiently spliced than the wild-type construct. These results indicate that, in the wild-type gene, precursor mRNA splicing is blocked in G0and that this block requires the E2F binding site. The data provide evidence for a coupling between stimulation of promoter activity and increased mRNA splicing in the mitogenic regulation of S phase-regulated genes. PMID- 9207023 TI - Purification and characterization of the RecF protein from Bacillus subtilis 168. AB - Genetic evidence suggests that the Bacillus subtilis recF gene product is involved in DNA repair and recombination. The RecF protein was overproduced and purified. NH2-terminal protein sequence analysis of RecF was consistent with the deduced amino acid sequence of the recF gene. The RecF protein (predicted molecular mass 42.3 kDa) bound single- and double-stranded DNA in a filter binding and in a gel retarding assay. The RecF-ssDNA or -dsDNA complex formation proceeds in the absence of nucleotide cofactors. RecF-ssDNA interaction is markedly stimulated by divalent cations. The apparent equilibrium constants of the RecF-DNA complexes are approximately 110-130 nM for both ssDNA and dsDNA. The binding reaction shows no cooperativity. The RecF protein does not physically interact with the RecR protein. Under our experimental conditions an ATPase activity was not associated with the purified RecF protein or with the RecF and RecR proteins. PMID- 9207024 TI - DNA containing 4'-thio-2'-deoxycytidine inhibits methylation by HhaI methyltransferase. AB - 4'-Thio-2'-deoxycytidine was synthesized as a 5'- protected phosphoramidite compatible with solid phase DNA synthesis. When incorporated as the target cytosine (C*) in the GC*GC recognition sequence for the DNA methyltransferase M. HhaI, methyl transfer was strongly inhibited. In contrast, these same oligonucleotides were normal substrates for the cognate restriction endonuclease R. HhaI and its isoschizomer R. Hin P1I. M. HhaI was able to bind both 4'-thio modified DNA and unmodified DNA to equivalent extents under equilibrium conditions. However, the presence of 4'-thio-2'-deoxycytidine decreased the half life of the complex by >10-fold. The crystal structure of a ternary complex of M. HhaI, AdoMet and DNA containing 4'-thio-2'-deoxycytidine was solved at 2.05 A resolution with a crystallographic R-factor of 0.186 and R-free of 0.231. The structure is not grossly different from previously solved ternary complexes containing M. HhaI, DNA and AdoHcy. The difference electron density suggests partial methylation at C5 of the flipped target 4'-thio-2'-deoxycytidine. The inhibitory effect of the 4'sulfur atom on enzymatic activity may be traced to perturbation of a step in the methylation reaction after DNA binding but prior to methyl transfer. This inhibitory effect can be partially overcome after a considerably long time in the crystal environment where the packing prevents complex dissociation and the target is accurately positioned within the active site. PMID- 9207025 TI - Site-specific introduction of functional groups into phosphodiester oligodeoxynucleotides and their thermal stability and nuclease-resistance properties. AB - We report here the site-specific introduction of functional groups into phosphodiester oligodeoxynucleotides (ODNs). ODNs containing both 5-( N aminohexyl)-carbamoyl-2'-deoxyuridine (H), which serves as a tether for the further conjugation of functional groups, and 5-(N,N-dimethylaminohexyl)carbamoyl 2'-deoxyuridine (D), which contributes to the thermal stability of the duplex and to the resistance to nucleolytic hydrolysis by nucleases, were synthesized. Functional groups such as folic acid and palmitic acid were site-specifically introduced into the terminus of the aminohexyl-linker of H. The thermal stability and resistance toward nuclease digestion of the modified ODNs were studied. We found that ODNs containing D and H formed stable duplexes with both the complementary DNA and RNA strands even when a bulky functional group such as folic acid, palmitic acid or cholesterol was attached to the terminus of the amino-linker. We also found that ODN analogues which contained D were more resistant to nucleolytic degradation by exo- and endonuclease than the unmodified ODN. Furthermore, duplexes formed by ODNs containing D and the complementary RNA could elicit RNase H activity. PMID- 9207026 TI - Hybridisation based DNA screening on peptide nucleic acid (PNA) oligomer arrays. AB - Arrays of up to some 1000 PNA oligomers of individual sequence were synthesised on polymer membranes using a robotic device originally designed for peptide synthesis. At approximately 96%, the stepwise synthesis efficiency was comparable to standard PNA synthesis procedures. Optionally, the individual, fully deprotected PNA oligomers could be removed from the support for further use, because an enzymatically cleavable but otherwise stable linker was used. Since PNA arrays could form powerful tools for hybridisation based DNA screening assays due to some favourable features of the PNA molecules, the hybridisation behaviour of DNA probes to PNA arrays was investigated for a precise understanding of PNA DNA interactions on solid support. Hybridisation followed the Watson-Crick base pairing rules with higher duplex stabilities than on corresponding DNA oligonucleotide sensors. Both the affinity and specificity of DNA hybridisation to the PNA oligomers depended on the hybridisation conditions more than expected. Successful discrimination between hybridisation to full complementary PNA sequences and truncated or mismatched versions was possible at salt concentrations down to 10 mM Na+and below, although an increasing tendency to unspecific DNA binding and few strong mismatch hybridisation events were observed. PMID- 9207027 TI - Location of the internal ribosome entry site in the 5' non-coding region of the immunoglobulin heavy-chain binding protein (BiP) mRNA: evidence for specific RNA protein interactions. AB - The 220 nucleotide 5'non-coding region (5'NCR) of the human immunoglobulin heavy chain binding protein (BiP) mRNA contains an internal ribosome entry site (IRES) that mediates the translation of the second cistron in a dicistronic mRNA in cultured mammalian cells. In this study, experiments are presented that locate the IRES immediately upstream of the start-site AUG codon in the BiP mRNA. Furthermore, crosslinking of thiouridine-labeled BiP IRES-containing RNA to cellular proteins identified the specific binding of two proteins, p60 and p95, to the 3'half of the BiP 5'NCR. Interestingly, both p60 and p95 bound also specifically to several viral IRES elements. This correlation suggests that p60 and p95 could have roles in internal initiation of cellular and viral IRES elements. PMID- 9207029 TI - New energy transfer dyes for DNA sequencing. AB - We have synthesized a set of four energy transfer dyes and demonstrated their use in automated DNA sequencing. The donor dyes are the 5- or 6-carboxy isomers of 4' aminomethylfluorescein and the acceptor dyes are a novel set of four 4,7-dichloro substituted rhodamine dyes which have narrower emission spectra than the standard, unsubstituted rhodamines. A rigid amino acid linker, 4 aminomethylbenzoic acid, was used to separate the dyes. The brightness of each dye in an automated sequencing instrument equipped with a dual line argon ion laser (488 and 514 nm excitation) was 2-2.5 times greater than the standard dye primers with a 2 times reduction in multicomponent noise. The overall improvement in signal-to-noise was 4- to 5-fold. The utility of the new dye set was demonstrated by sequencing of a BAC DNA with an 80 kb insert. Measurement of the extinction coefficients and the relative quantum yields of the dichlororhodamine components of the energy transfer dyes showed their values were reduced by 20-25% compared with the dichlororhodamine dyes alone. PMID- 9207028 TI - MS2 coat protein mutants which bind Qbeta RNA. AB - The coat proteins of the RNA phages MS2 and Qbetaare structurally homologous, yet they specifically bind different RNA structures. In an effort to identify the basis of RNA binding specificity we sought to isolate mutants that convert MS2 coat protein to the RNA binding specificity of Qbeta. A library of mutations was created which selectively substitutes amino acids within the RNA binding site. Genetic selection for the ability to repress translation from the Qbetatranslational operator led to the isolation of several MS2 mutants that acquired binding activity for QbetaRNA. Some of these also had reduced abilities to repress translation from the MS2 translational operator. These changes in RNA binding specificity were the results of substitutions of amino acid residues 87 and 89. Additional codon- directed mutagenesis experiments confirmed earlier results showing that the identity of Asn87 is important for specific binding of MS2 RNA. Glu89, on the other hand, is not required for recognition of MS2 RNA, but prevents binding of QbetaRNA. PMID- 9207030 TI - Complementation of the DNA repair-deficient swi10 mutant of fission yeast by the human ERCC1 gene. AB - In human cells DNA damage caused by UV light is mainly repaired by the nucleotide excision repair pathway. This mechanism involves dual incisions on both sides of the damage catalyzed by two nucleases. In mammalian cells XPG cleaves 3' of the DNA lesion while the ERCC1-XPF complex makes the 5' incision. The amino acid sequence of the human excision repair protein ERCC1 is homologous with the fission yeast Swi10 protein. In order to test whether these proteins are functional homologues, we overexpressed the human gene in a Schizosaccharomyces pombe swi10 mutant. A swi10 mutation has a pleiotropic effect: it reduces the frequency of mating type switching (a mitotic transposition event from a silent cassette into the expression site) and causes increased UV sensitivity. We found that the full-length ERCC1 gene only complements the transposition defect of the fission yeast mutant, while a C-terminal truncated ERCC1 protein also restores the DNA repair capacity of the yeast cells. Using the two-hybrid system of Saccharomyces cerevisiae we show that only the truncated human ERCC1 protein is able to interact with the S . pombe Rad16 protein, which is the fission yeast homologue of human XPF. This is the first example yet known that a human gene can correct a yeast mutation in nucleotide excision repair. PMID- 9207031 TI - Segmental genomic replacement by Cre-mediated recombination: genotoxic stress activation of the p53 promoter in single-copy transformants. AB - Genotoxic stress results in transcriptional activation of the p53 promoter. To gain more detailed information on genotoxic induction of the p53 promoter at a uniform genomic locus, we have developed an efficient strategy for replacing a defined genomic segment in mouse NIH 3T3 cells with exogenous transfected DNA using a 'double lox' targeting strategy mediated by Cre DNA recombinase. The strategy utilizes a pair of heterospecific lox sites engineered both into the genome and onto the targeting DNA. This allows direct replacement of genomic DNA by a Cre-catalyzed double crossover event. p53-CAT reporter constructs were site specifically placed into the genomic target 20-fold more efficiently by double lox recombination than by Cre-mediated single crossover insertional recombination, and the absolute frequency of site-specific double lox targeting exceeded the frequency of transformation due to random illegitimate recombination of transfected DNA into the genome. Resulting targeted single-copy integrants of the p53-CAT reporter show strong genotoxic induction by mitomycin C, and a dynamic range of induction that exceeds that seen in transient transfection assays. The double lox strategy is generally applicable to Cre-mediated genomic targeting in any cell and should be of particular utility in the site-specific targeting of DNA into embryonic stem (ES) cells for the production of gene modified mice. PMID- 9207032 TI - Determinants of a protein-induced RNA switch in the large domain of signal recognition particle identified by systematic-site directed mutagenesis. AB - Signal recognition particle (SRP) is a ribonucleoprotein complex that associates with ribosomes to promote the co-translational translocation of proteins across biological membranes. Human SRP RNA molecules exist in two distinct conformations, SR-A and SR-B, which may exchange during the assembly of the particle or could play a functional role in the SRP cycle. We have used systematic site-directed mutagenesis of the SRP RNA to determine the electrophoretic mobilities of altered RNA molecules, and we have identified the nucleotides that avert the formation of the two conformers. The conformer behavior of the various RNAs was addressed quantitatively by calculating a value zeta as an indicator of conformational variability. Single loose A-like forms were induced by changes in helix 5 [nucleotides (nt) at positions 111-128 or 222 231], helix 6 (nt at positions 141-150) and helix 7 (nt at position 169 and 170), whereas other mutations in helix 6 and helix 8 preserved the conformational variability of the mutant RNA molecules. The more compact B-like form was induced only when a small region (129-CAAUAU-134), located in the 5'-proximal portion of helix 6, was altered. Since this region is part of the binding site for SRP19, we suggest that protein SRP19 uses nucleotides at 129-134 to trigger the formation of the favored SR-B-form, and thus directs an early step in the hierarchical assembly of the large SRP domain. PMID- 9207033 TI - Footprint analysis of the bsp RI DNA methyltransferase-DNA interaction. AB - The interaction between the GGCC-specific Bsp RI DNA methyltransferase (M. Bsp RI) and substrate DNA was studied with footprinting techniques using a DNA fragment that was unmodified on both strands. Footprinting with DNase I revealed an approximately 14 bp protected region. Footprinting with dimethylsulfate detected major groove interactions with the guanine bases of the recognition sequence. Reaction with 1,10-phenanthroline-copper did not show protection, suggesting that minor groove interactions play little role in sequence-specific recognition by M. Bsp RI. Hydroxyl radical footprinting revealed a protected stretch of 6 nt. The hydroxyl radical footprint of M. Bsp RI differs markedly from the the footprint reported for the Hha I and Sss I methyltransferases. The pattern of protection from dimethylsulfate and hydroxyl radicals suggests that the interactions of M. Bsp RI with DNA are similar to those detected in the co crystal structure of the Hae III methyltransferase. PMID- 9207034 TI - Cysteine 50 of the POU H domain determines the range of targets recognized by POU proteins. AB - The best target of POU proteins (Oct-1, Oct-2) is an octamer sequence ATGCAAAT. POU proteins also recognize, with weaker affinity, the TAAT-like targets of another group of regulatory factors, the homeoproteins. Up to now, it has not been known why Cys50 of the POUHdomain is absolutely conserved in contrast to that in homeoproteins. To assess the importance of Cys50 in determining the binding specificity of POU proteins, all possible amino acids were substituted for Cys at position 50, and the resulting mutants were tested with probes containing octamer (ATGCAAATNN) or homeospecific binding sites. Only the wild type POU was shown to adequately discriminate between the octamer and homeospecific sites, and the protein affinity was only slightly affected by the nucleotide sequence flanking the octamer at the 3'-end. Any amino acid substitution at position 50 resulted in the mutant protein binding efficiently both to the octamer and the TAAT-like sequences. Moreover, in this case the 3' flanking sequences influenced the binding to a much greater extent. PMID- 9207035 TI - Plant ribosome shunting in vitro. AB - It has been proposed that cauliflower mosaic virus 35S RNA with its 600 nt long leader uses an unusual translation process (the translational shunt). A wheat germ in vitro translation assay was used to improve the study of this mechanism. Deletions, the introduction of stable stem-loop structures, and the inhibitory effect of antisense oligonucleotides on gene expression were used to determine the roles of various parts of the leader. It was found that the 5'- and 3'-ends of the leader are absolutely required for translation whereas the middle part is apparently dispensable. These results confirm the data already reported from transient expression experiments with protoplasts. However, the in vitro data suggest in contrast to protoplast experiments that only two relatively short regions at both ends, approximately 100 nt each, are required. The in vitro system provides tools for further studying the shunt model at the molecular level and for examining the involvement of proteins in this mechanism. Shunting was also found to occur with the rice tungro bacilliform virus leader. As wheat is neither a host plant of cauliflower mosaic virus nor rice tungro bacilliform virus, the shunt seems to be host independent, a finding that deviates from earlier studies in protoplasts. PMID- 9207036 TI - Transcription increases the deletion frequency of long CTG.CAG triplet repeats from plasmids in Escherichia coli. AB - Induction of transcription into long CTG.CAG repeats contained on plasmids in Escherichia coli is shown to increase the frequency of deletions within the repeat sequences. This elevated genetic instability was detected because active transcription into the triplet repeat influenced the growth transitions of the host cell, allowing advantageous growth for cells harboring plasmids with deleted repeat sequences. The variety of deletion products observed in separate cultures suggests that transcription altered the metabolism of the DNA in a manner that produced random length changes in the repeat sequence. For cultures containing plasmids without active transcription into the triplet repeat, or those maintained in exponential growth, deletions occurred within the repeat at a lower frequency (5-20-fold lower). In these incubations the extent of deletions was proportional to the number of cell divisions and many repeat lengths were observed within each culture, suggesting that the decrease in average repeat length at long incubation times was due to multiple small deletions. These observations show that deletions within long CTG.CAG repeats contained on plasmids in E.coli occur via more than one pathway and their level of genetic instability is altered by the enzymatic processes occurring upon the DNA. PMID- 9207037 TI - The conserved 5'-untranslated leader of Spi-1 (PU.1) mRNA is highly structured and potently inhibits translation in vitro but not in vivo. AB - The transcription factor Spi-1 (PU.1) has a central role in regulating myeloid gene expression during hematopoietic development and its overexpression has been implicated in erythroleukemic transformation. Thus regulation of Spi-1 expression has broad significance for hematopoietic development. A comparison of human and murine cDNA sequences demonstrates that the 5'-untranslated region (5'-UTR) of Spi-1 mRNA is as highly conserved as the coding region (87% identical), suggesting that this sequence may be involved in regulating expression of this protein. The experiments presented in this manuscript provide evidence that the 5'-UTR of Spi-1 contains extensive secondary structure, including three stem loops that precede the AUG codon. Analysis of the in vitro transcribed Spi-1 5' UTR by partial nuclease digestion sensitivity is consistent with the existence of two of these stem-loops. The 5'-UTR decreased translation of Spi-1 transcripts in reticuloctye lysates 8- to 10-fold. A series of partial deletions of the 5'-UTR identified the sequence corresponding to the stem-loop most proximal to the initiating AUG codon as sufficient for inhibition of translation. However, the effect of the 5'-UTR on translation in vivo was negligible and resulted in only a slight reduction in the number of ribosomes that became associated with the mRNA. Further, this sequence had no affect on expression of luciferase. The disparity between in vivo and in vitro effects, coupled with the observation that endogenous Spi-1 mRNA is wholly associated with polysomes in MEL cells, suggests that additional cellular mechanisms contribute to regulation of Spi-1 expression in these cells or that conservation of these sequences serves a function that is independent of translation. PMID- 9207038 TI - Effect of in vitro promoter methylation and CGG repeat expansion on FMR-1 expression. AB - Fragile X syndrome is associated with a CGG repeat expansion in the 5' untranslated region of the FMR-1 gene. Within the FMR-1 promoter a CpG island is frequently methylated in fragile X patients. To identify the effect of methylation on FMR-1 expression, we transfected methylated and unmethylated constructs containing the FMR-1 promoter in front of the CAT gene (pFXCAT) into COS-1 cells. No difference between methylated and unmethylated DNA was observed initially, whereas reduced CAT mRNA levels were observed 48 h post-transfection of the methylated construct and increased CAT activity from unmethylated DNA was observed at 72 h. To determine the effect of a CGG repeat expansion on gene expression, we inserted >200 CGG repeats between the SV40 promoter and the CAT gene (pSV2CAT). A 3-fold reduction in CAT activity was observed 24-48 h post transfection. To study the correlation between CGG repeat expansion and FMR-1 transcription, we inserted 200 CGG trinucleotide repeats into the pFXCAT construct. Only a slight difference in mRNA levels was found between cells transfected with pFX(CGG)200CAT or pFXCAT, but a complete lack of CAT activity was observed with introduction of the repeat. We conclude that a moderate size repeat markedly reduces translation. We propose that the presence of a repeat expansion per se is the major factor influencing FMR-1 function in fragile X syndrome. PMID- 9207039 TI - Different DNA contact schemes are used by two winged helix proteins to recognize a DNA binding sequence. AB - The hepatocyte nuclear factor 3 (HNF-3)/fork head (fkh) family contains a large number of transcription factors which recognize divergent DNA sequences via a winged-helix binding motif. In this report we present studies on the DNA binding properties of winged-helix HNF-3/fkh homologues 1 (HFH-1) and 2 (HFH-2) which recognize a shared DNA binding site with different affinities. To explore how HFH 1 and HFH-2 proteins recognize this DNA binding sequence, the binding affinities of these two HFH proteins toward a series of DNA sites containing a single strand trimer loop insertion at different positions were studied. This insertion induces a bend of approximately 80 degrees in the DNA binding site (prebending). HFH-1 and HFH-2 were shown to recognize DNA sites prebent at many nucleotide positions on both strands of the DNA sequence. Both HFH-1 and HFH-2 were more sensitive to mismatch insertions on the sense strand of the DNA binding site, especially within the AAAATAAC sequence. Our data suggest that the recognition helix (helix 3) recognizes the AAAATAAC sequence and that the helix 3/DNA interaction results in bending of the DNA which narrows the major groove in the AAAATAAC sequence. Furthermore, the binding affinities of HFH-1 and HFH-2 toward DNA binding sites with base-pair reversion in the AAAATAAC sequence was also investigated. Different patterns of response from HFH-1 and HFH-2 to both prebent and base-pair reverted binding sites was observed. Our results demonstrate that even two highly conserved members of the winged-helix family may contact the same DNA sequence differently. PMID- 9207040 TI - Synthesis and radioiodination of a stannyl oligodeoxyribonucleotide. AB - Synthesis and radioiodination of a stannyl oligodeoxyribonucleotide were undertaken to evaluate a gamma ray emitting ODN ligand for thrombus imaging in vivo . Synthesis of the ODN was based on modified automatedbeta-cyanoethyl phosphoramidite chemistry with an organotin nucleoside (dU*) coupled to a thrombin binding aptamer sequence to give d(U*GGTTGGTGTGGTTGG). The synthesis accommodated dU*, which is destannylated by iodine or acids. Fourteen standard synthesis cycles were followed by one 'stannyl synthesis cycle', distinguished by Fmoc protection, omission of capping, oxidation by an organic peroxide and cleavage by ammonium hydroxide. The organotin nucleoside phosphoramidite {5' [fluorenylmethoxycarbonyl]-5-(E)-[2-tri-n -butylstannylvinyl]-2'-deoxyuridine-3' (2-cyanoethyl N,N-diisopropyl phosphoramidite)} was prepared from 5-iodo-2' deoxyuridine. A customized mild rapid workup included deprotection with methylamine, and reverse phase HPLC with CH3CN/triethylammonium bicarbonate. Pure stannyl ODN was highly retained by reverse phase HPLC. Radioiodination of stannyl ODN (100 microg) provided 123I-labeling yields up to 97%. Five alternative oxidants were effective. High specific activity [123I]- ODN (15 000 Ci/mmol) was recovered, separated from unlabeled isomers. Excellent reverse phase HPLC resolution of ODN isomers (alternatively I, Cl, H or Br in vinyl deoxyuridine) was essential. The affinity of the iodovinyl aptamer analog (Kd = 36 nM) for human alpha-thrombin was similar to the native aptamer (Kd = 45 nM). PMID- 9207041 TI - In vitro selection of RNAs that bind to the human immunodeficiency virus type-1 gag polyprotein. AB - RNA ligands that bind to the human immunodeficiency virus type-1 (HIV-1) gag polyprotein with 10(-9) M affinity were isolated from a complex pool of RNAs using an in vitro selection method. The ligands bind to two different regions within gag, either to the matrix protein or to the nucleocapsid protein. Binding of a matrix ligand to gag did not interfere with the binding of a nucleocapsid ligand, and binding of a nucleocapsid ligand to gag did not interfere with the binding of a matrix ligand. However, binding of a nucleocapsid ligand to gag did interfere with binding of an RNA containing the HIV-1 RNA packaging element (psi), even though the sequence of the nucleocapsid ligand is not similar topsi. The minimal sequences required for the ligands to bind to matrix or nucleocapsid were determined. Minimal nucleocapsid ligands are predicted to form a stem-loop structure that has a self-complementary sequence at one end. Minimal matrix ligands are predicted to form a different stem-loop structure that has a CAARU loop sequence. The properties of these RNA ligands may provide tools for studying RNA interactions with matrix and nucleocapsid, and a novel method for inhibiting HIV replication. PMID- 9207042 TI - Overexpression in COS cells of p50, the major core protein associated with mRNA, results in translation inhibition. AB - p50, the major core protein of messenger ribonucleoprotein particles (mRNPs) in the cytoplasm of somatic mammalian cells, has been characterized previously as a member of the Y-box binding transcription factor family of proteins (YB-protein) by both high structural homology and ability to bind specifically the Y-box sequence in double-stranded DNA. YB proteins are present in a whole range of cell types and some have been identified as germ-specific cytoplasmic proteins masking stored mRNA from translation. Western blot analysis of the distribution of p50 in subcellular fractions of COS-1 cells shows that p50 is a cytoplasmic protein quantitatively associated with mRNA, both in polyribosomes and in free mRNPs. The level of p50 in COS-1 cells determined by Western immunoblotting is 0.10% of total protein, which is nearly equimolar to that of ribosomes and is approximately 5-10-fold higher than the mRNA level. Transient transfection of COS 1 cells with a p50-expressing vector results in a dramatic inhibition of protein synthesis. A control transfection with a vector expressing a frameshift mutant of p50 does not cause translation inhibition. Therefore the increase in p50 protein level is responsible for the inhibitory effect in these cells. PMID- 9207044 TI - Stability, specificity and fluorescence brightness of multiply-labeled fluorescent DNA probes. AB - In this work, we studied the fluorescence and hybridization of multiply-labeled DNA probes which have the hydrophilic fluorophore 1-(straightepsilon carboxypentynyl)-1'-ethyl- 3,3,3', 3'-tetramethylindocarbocyanine-5,5' disulfonate (Cy3) attached via either a short or long linker at the C-5 position of deoxyuridine. We describe the effects of labeling density, fluorophore charge and linker length upon five properties of the probe: fluorescence intensity, the change in fluorescence upon duplex formation, the quantum yield of fluorescence (Phif), probe-target stability and specificity. For the hydrophilic dye Cy3, we have demonstrated that the fluorescence intensity andPhifare maximized when labeling every 6th base using the long linker. With a less hydrophilic dye, a labeling density this high could not be achieved without serious quenching of the fluorescence. The target specificity of multiply-labeled DNA probes was just as high as compared to the unmodified control probe, however, a less stable probe target duplex is formed that exhibits a lower melting temperature. A mechanism that accounts for this destabilization is proposed which is consistent with our data. It involves dye-dye and dye-nucleotide interactions which appear to stabilize a single-stranded conformation of the probe. PMID- 9207043 TI - Analysis of enhancer function of the HS-40 core sequence of the human alpha globin cluster. AB - HS-40 is the major regulatory element of the human alpha-globin locus, located 40 kb upstream of the zeta-globin gene. To test for potential interactions between HS-40 and the beta- or the gamma-globin gene promoters in stable transfection assays, the HS-40 core sequence was cloned upstream of either the beta promoter or the gamma promoter driving the neomycin phosphotransferase gene and enhancer activity was measured using a colony assay. In K562 or in MEL cells, enhancer activity of HS-40 was higher than that of the individual core sequences of the DNase I hypersensitive sites (HS) of the beta-globin locus control region (LCR), and approximately 60% of the enhancer activity of a 2.5 kb microLCR, which contains the core elements of DNase I hypersensitive sites 1-4. In contrast to the synergistic interaction between the DNase I hypersensitive sites of beta locus LCR, combination of HS-40 with these DNase I hypersensitive sites failed to display cooperativity in K562 cells and inhibited enhancer function in MEL cells. Inhibition of enhancer function was also observed when two copies of the HS-40 were arranged tandemly. We conclude that the core element of HS-40 (i) is a powerful enhancer of gamma- and beta-globin gene expression, (ii) in contrast to other classical enhancers, acts best as a single copy, (iii) does not cooperate with the regulatory elements of the beta-globin locus control region. PMID- 9207046 TI - A universal procedure for primer labelling of amplicons. AB - Detection and visualisation of nucleic acids is integral to genome analyses. Exponential amplification procedures have provided the means for the manipulation of nucleic acid sequences, which were otherwise inaccessible. We describe the development and application of a universal method for the labelling of any PCR product using a single end-labelled primer. Amplification was performed in a single reaction with the resulting amplicon labelled to a high specific activity. The method was adapted to a wide range of PCRs and significantly reduced the expense of such analyses. PMID- 9207045 TI - Proto-oncogene Sno expression, alternative isoforms and immediate early serum response. AB - The mouse Sno gene, a Ski proto-oncogene homolog, expresses two isoforms, SnoN and SnoN2 (also called sno -dE3), which differ from each other in a location downstream from the site of alternative splicing previously described in the human SNO gene. SnoN2 is missing a 138 nt coding segment present in mouse SnoN and human SNON . We have cloned and sequenced the human ortholog of mouse SnoN2 , the existence of which was predicted from conservation of the alternative splice donor site that produces the SnoN2 isoform. Mouse SnoN2 and SnoN are expressed throughout embryonic development, in neonatal muscle and in many adult tissues. SnoN2 is the major species in most tissues, but SnoN and SnoN2 are expressed at approximately equal levels in brain. In human tissues, SNON2 is the less abundantly expressed isoform. Expression of mouse SnoN and SnoN2 mRNAs is induced with immediate early kinetics upon serum stimulation of quiescent fibroblasts, even in the presence of the protein synthesis inhibitor cycloheximide, while Ski is not. Interestingly, although both isoforms of Sno are induced, SnoN2 induction is much higher than SnoN . These data are consistent with a role for Sno in the response to proliferation stimuli. PMID- 9207047 TI - Efficient recovery and regeneration of integrated retroviruses. AB - We report a rapid and efficient PCR-based rescue procedure for integrated recombinant retroviruses. Full-length proviral DNA is amplified by long-range PCR using a pair of primers derived from the long terminal repeats (LTR), and virus is regenerated by transfecting retrovirus-packaging cells with the PCR-derived provirus. The viral yield from the PCR product is similar to that from the retroviral plasmid vector, and the representation of different inserts is accurately maintained in the recovered retroviral population. This procedure is suitable for expression cloning from retroviral libraries and should be applicable to the analysis of natural retrovirus populations. PMID- 9207048 TI - Avoidance of sulfur loss during ammonia treatment of oligonucleotide phosphorothioates. AB - Sulfur loss during the unblocking of phosphorothioate analogues of oligonucleotides with concentrated aqueous ammonia can be completely suppressed by the addition of 2-mercaptoethanol. PMID- 9207049 TI - SYBR Green I staining of pulsed field agarose gels is a sensitive and inexpensive way of quantitating DNA double-strand breaks in mammalian cells. AB - Pulsed field gel electrophoresis (PFGE) is widely used to measure DNA double strand breaks (dsb). The DNA of cultured cells can be prelabelled with radioactivity, which helps greatly in detection and quantitation of DNA dsb. However, this approach cannot be used with non-cycling cells from biopsy material. We describe a method which uses SYBR Green I to stain DNA in dried agarose gels. DNA is detected and analysed using readily available camera equipment and image analysis software. This method is as sensitive as [3H]thymidine prelabelling of cells and allows DNA dsb to be measured simply and economically in non-cycling cells. PMID- 9207050 TI - A homogeneous method to quantify mRNA levels: a hybridization of RNase protection and scintillation proximity assay technologies. AB - A novel method to measure mRNA levels has been developed by combining the detection capabilities of RNase protection (RPA) with the quantification advantages of scintillation proximity assay (SPA) technology. Sample processing is reduced to the addition of a single reagent post RNase digestion. As a model system, the inducible expression of rat apolipoprotein-A1 mRNA has been measured by both traditional gel-based RPAs and the SPA-based RPA assay. Results demonstrate that the ribonuclease protection proximity assay (RiPPA) faithfully reproduces the gel-based results and is at least as sensitive as many existing methods. PMID- 9207051 TI - CCR: a rapid and simple approach for mutation detection. AB - We describe a simple approach for detecting known mutations in genomic DNA. The strategy entails a DNA amplification reaction that combines the use of thermostable DNA polymerase and ligase, and that has been designated the Combined Chain Reaction (CCR). CCR consists of four phases: denaturation, annealing, elongation and ligation. Unlike most PCR-based mutation detection systems it relies on mismatch between primer and template at the primer 5'ends. It is rapid and simple, and requires neither the use of radioactivity, nor polyacrylamide gel electrophoresis, nor autoradiography for mutation detection at the single base pair level. PMID- 9207053 TI - Hepatitis B virus, the vaccine, and the control of primary cancer of the liver. PMID- 9207054 TI - Natural and unnatural answers to evolutionary questions. PMID- 9207055 TI - Contacting the protein folding funnel with NMR. PMID- 9207057 TI - BRCA1 is a cell cycle-regulated nuclear phosphoprotein. AB - We have characterized the BRCA1 gene product by using four polyclonal antibodies raised against peptides from four different regions of the protein. The antibodies specifically recognize an approximately 220-kDa BRCA1 protein that is predominantly expressed in the nucleus of both normal and neoplastic breast cancer cells. It is a serine phosphoprotein that undergoes hyperphosphorylation during late G1 and S phases of the cell cycle and is transiently dephosphorylated early after M phase. We propose that BRCA1 is a phosphoprotein that alters in a qualitative and quantitative manner during cell cycle progression. PMID- 9207058 TI - The first step of aminoacylation at the atomic level in histidyl-tRNA synthetase. AB - The crystal structure of an enzyme-substrate complex with histidyl-tRNA synthetase from Escherichia coli, ATP, and the amino acid analog histidinol is described and compared with the previously obtained enzyme-product complex with histidyl-adenylate. An active site arginine, Arg-259, unique to all histidyl-tRNA synthetases, plays the role of the catalytic magnesium ion seen in seryl-tRNA synthetase. When Arg-259 is substituted with histidine, the apparent second order rate constant (kcat/Km) for the pyrophosphate exchange reaction and the aminoacylation reaction decreases 1,000-fold and 500-fold, respectively. Crystals soaked with MnCl2 reveal the existence of two metal binding sites between beta- and gamma-phosphates; these sites appear to stabilize the conformation of the pyrophosphate. The use of both conserved metal ions and arginine in phosphoryl transfer provides evidence of significant early functional divergence of class II aminoacyl-tRNA synthetases. PMID- 9207060 TI - Identification of the yeast 20S proteasome catalytic centers and subunit interactions required for active-site formation. AB - The proteasome is responsible for degradation of substrates of the ubiquitin pathway. 20S proteasomes are cylindrical particles with subunits arranged in a stack of four heptameric rings. The outer rings are composed of alpha subunits, and the inner rings are composed of beta subunits. A well-characterized archaeal proteasome has a single type of each subunit, and the N-terminal threonine of the beta subunit is the active-site nucleophile. Yeast proteasomes have seven different beta subunits and exhibit several distinct peptidase activities, which were proposed to derive from disparate active sites. We show that mutating the N terminal threonine in the yeast Pup1 beta subunit eliminates cleavage after basic residues in peptide substrates, and mutating the corresponding threonine of Pre3 prevents cleavage after acidic residues. Surprisingly, neither mutation has a strong effect on cell growth, and they have at most minor effects on ubiquitin dependent proteolysis. We show that Pup1 interacts with Pup3 in each beta subunit ring. Our data reveal that different proteasome active sites contribute very differently to protein breakdown in vivo, that contacts between particular subunits in each beta subunit ring are critical for active-site formation, and that active sites in archaea and different eukaryotes are highly similar. PMID- 9207059 TI - RAP74 induces promoter contacts by RNA polymerase II upstream and downstream of a DNA bend centered on the TATA box. AB - RAP74, the large subunit of transcription factor IIF, associates with a preinitiation complex containing RNA polymerase II (pol II) and other general initiation factors. We have mapped the location of RAP74 in close proximity to promoter DNA at similar distances both upstream and downstream of a DNA bend centered on the TATA box. Binding of RAP74 induces a conformational change that affects the position of pol II relative to that of the DNA. This reorganization of the preinitiation complex minimally requires the N-terminal region of RAP74 containing both its RAP30-binding domain and another region necessary for accurate transcription in vitro. We propose a role for RAP74 in controlling the topological organization of the pol II preinitiation complex. PMID- 9207061 TI - X-ray analysis of azido-thymidine diphosphate binding to nucleoside diphosphate kinase. AB - To be effective as antiviral agent, AZT (3'-azido-3'-deoxythymidine) must be converted to a triphosphate derivative by cellular kinases. The conversion is inefficient and, to understand why AZT diphosphate is a poor substrate of nucleoside diphosphate (NDP) kinase, we determined a 2.3-A x-ray structure of a complex with the N119A point mutant of Dictyostelium NDP kinase. It shows that the analog binds at the same site and, except for the sugar ring pucker which is different, binds in the same way as the natural substrate thymidine diphosphate. However, the azido group that replaces the 3'OH of the deoxyribose in AZT displaces a lysine side chain involved in catalysis. Moreover, it is unable to make an internal hydrogen bond to the oxygen bridging the beta- and gamma phosphate, which plays an important part in phosphate transfer. PMID- 9207062 TI - Interaction of human apurinic endonuclease and DNA polymerase beta in the base excision repair pathway. AB - Mutagenic abasic (AP) sites are generated directly by DNA-damaging agents or by DNA glycosylases acting in base excision repair. AP sites are corrected via incision by AP endonucleases, removal of deoxyribose 5-phosphate, repair synthesis, and ligation. Mammalian DNA polymerase beta (Polbeta) carries out most base excision repair synthesis and also can excise deoxyribose 5-phosphate after AP endonuclease incision. Yeast two-hybrid analysis now indicates protein-protein contact between Polbeta and human AP endonuclease (Ape protein). In vitro, binding of Ape protein to uncleaved AP sites loads Polbeta into a ternary complex with Ape and the AP-DNA. After incision by Ape, only Polbeta exhibits stable DNA binding. Kinetic experiments indicated that Ape accelerates the excision of 5' terminal deoxyribose 5-phosphate by Polbeta. Thus, the two central players of the base excision repair pathway are coordinated in sequential reactions. PMID- 9207063 TI - ETS target genes: identification of egr1 as a target by RNA differential display and whole genome PCR techniques. AB - ETS transcription factors play important roles in hematopoiesis, angiogenesis, and organogenesis during murine development. The ETS genes also have a role in neoplasia, for example in Ewing's sarcomas and retrovirally induced cancers. The ETS genes encode transcription factors that bind to specific DNA sequences and activate transcription of various cellular and viral genes. To isolate novel ETS target genes, we used two approaches. In the first approach, we isolated genes by the RNA differential display technique. Previously, we have shown that the overexpression of ETS1 and ETS2 genes effects transformation of NIH 3T3 cells and specific transformants produce high levels of the ETS proteins. To isolate ETS1 and ETS2 responsive genes in these transformed cells, we prepared RNA from ETS1, ETS2 transformants, and normal NIH 3T3 cell lines and converted it into cDNA. This cDNA was amplified by PCR and displayed on sequencing gels. The differentially displayed bands were subcloned into plasmid vectors. By Northern blot analysis, several clones showed differential patterns of mRNA expression in the NIH 3T3-, ETS1-, and ETS2-expressing cell lines. Sixteen clones were analyzed by DNA sequence analysis, and 13 of them appeared to be unique because their DNA sequences did not match with any of the known genes present in the gene bank. Three known genes were found to be identical to the CArG box binding factor, phospholipase A2-activating protein, and early growth response 1 (Egr1) genes. In the second approach, to isolate ETS target promoters directly, we performed ETS1 binding with MboI-cleaved genomic DNA in the presence of a specific mAb followed by whole genome PCR. The immune complex-bound ETS binding sites containing DNA fragments were amplified and subcloned into pBluescript and subjected to DNA sequence and computer analysis. We found that, of a large number of clones isolated, 43 represented unique sequences not previously identified. Three clones turned out to contain regulatory sequences derived from human serglycin, preproapolipoprotein C II, and Egr1 genes. The ETS binding sites derived from these three regulatory sequences showed specific binding with recombinant ETS proteins. Of interest, Egr1 was identified by both of these techniques, suggesting strongly that it is indeed an ETS target gene. PMID- 9207064 TI - The primary fibrin polymerization pocket: three-dimensional structure of a 30-kDa C-terminal gamma chain fragment complexed with the peptide Gly-Pro-Arg-Pro. AB - After vascular injury, a cascade of serine protease activations leads to the conversion of the soluble fibrinogen molecule into fibrin. The fibrin monomers then polymerize spontaneously and noncovalently to form a fibrin gel. The primary interaction of this polymerization reaction is between the newly exposed N terminal Gly-Pro-Arg sequence of the alpha chain of one fibrin molecule and the C terminal region of a gamma chain of an adjacent fibrin(ogen) molecule. In this report, the polymerization pocket has been identified by determining the crystal structure of a 30-kDa C-terminal fragment of the fibrin(ogen) gamma chain complexed with the peptide Gly-Pro-Arg-Pro. This peptide mimics the N terminus of the alpha chain of fibrin. The conformational change in the protein upon binding the peptide is subtle, with electrostatic interactions primarily mediating the association. This is consistent with biophysical experiments carried out over the last 50 years on this fundamental polymerization reaction. PMID- 9207065 TI - Detection of residue contacts in a protein folding intermediate. AB - Protein folding can be described in terms of the development of specific contacts between residues as a highly disordered polypeptide chain converts into the native state. Here we describe an NMR based strategy designed to detect such contacts by observation of nuclear Overhauser effects (NOEs). Experiments with alpha-lactalbumin reveal the existence of extensive NOEs between aromatic and aliphatic protons in the archetypal molten globule formed by this protein at low pH. Analysis of their time development provides direct evidence for near-native compactness of this state. Through a rapid refolding procedure the NOE intensity can be transferred efficiently into the resolved and assigned spectrum of the native state. This demonstrates the viability of using this approach to map out time-averaged interactions between residues in a partially folded protein. PMID- 9207066 TI - Interaction between replication protein A and p53 is disrupted after UV damage in a DNA repair-dependent manner. AB - Replication protein A (RPA) is required for both DNA replication and nucleotide excision repair. Previous studies have shown that RPA interacts with the tumor suppressor p53. Herein, we have mapped a 20-amino acid region in the N-terminal part of p53 that is essential for its binding to RPA. This region is distinct from the minimal activation domain of p53 previously identified. We also demonstrate that UV radiation of cells greatly reduces the ability of RPA to bind to p53. Interestingly, damage-induced hyperphosphorylated RPA does not associate with p53. Furthermore, down-regulation of the RPA/p53 interaction is dependent upon the capability of cells to perform global genome repair. On the basis of these data, we propose that RPA may participate in the coordination of DNA repair with the p53-dependent checkpoint control by sensing UV damage and releasing p53 to activate its downstream targets. PMID- 9207067 TI - Vascular endothelial growth factor: crystal structure and functional mapping of the kinase domain receptor binding site. AB - Vascular endothelial growth factor (VEGF) is a homodimeric member of the cystine knot family of growth factors, with limited sequence homology to platelet-derived growth factor (PDGF) and transforming growth factor beta2 (TGF-beta). We have determined its crystal structure at a resolution of 2.5 A, and identified its kinase domain receptor (KDR) binding site using mutational analysis. Overall, the VEGF monomer resembles that of PDGF, but its N-terminal segment is helical rather than extended. The dimerization mode of VEGF is similar to that of PDGF and very different from that of TGF-beta. Mutational analysis of VEGF reveals that symmetrical binding sites for KDR are located at each pole of the VEGF homodimer. Each site contains two functional "hot spots" composed of binding determinants presented across the subunit interface. The two most important determinants are located within the largest hot spot on a short, three-stranded sheet that is conserved in PDGF and TGF-beta. Functional analysis of the binding epitopes for two receptor-blocking antibodies reveal different binding determinants near each of the KDR binding hot spots. PMID- 9207068 TI - Multiple loop structures critical for ligand binding of the integrin alpha4 subunit in the upper face of the beta-propeller mode 1. AB - A non-I-domain integrin, alpha4beta1, recognizes vascular cell adhesion molecule 1 (VCAM-1) and the IIICS portion of fibronectin. To localize regions of alpha4 critical for ligand binding, we swapped several predicted loops within or near the putative ligand-binding site of alpha4 (which spans repeats 2-5 of the seven N-terminal repeats) with the corresponding regions of alpha5. Swapping residues 112-131 in repeat 2, or residues 237-247 in repeat 4, completely blocked adhesion to immobilized VCAM-1 and connecting segment 1 (CS-1) peptide. However, swapping residues 40-52 in repeat 1, residues 151-164 in repeat 3, or residues 282-288 (which contain a putative cation binding motif) in repeat 5 did not affect or only slightly reduced adhesion to these ligands. The binding of several function blocking antibodies is blocked by swapping residues 112-131, 151-164, and 186-191 (which contain previously identified residues critical for ligand binding, Tyr 187 and Gly-190). These results are consistent with the recently published beta propeller folding model of the integrin alpha4 subunit [Springer, T. A. (1997) Proc. Natl. Acad. Sci. USA 94, 65-72], in which seven four-stranded beta-sheets are arranged in a torus around a pseudosymmetric axis. The regions of alpha4 critical for ligand binding are adjacent to each other and are located in the upper face, the predicted ligand-binding site, of the beta-propeller model, although they are not adjacent in the primary structure. PMID- 9207069 TI - High-affinity binding of the Drosophila Numb phosphotyrosine-binding domain to peptides containing a Gly-Pro-(p)Tyr motif. AB - The phosphotyrosine-binding (PTB) domain is a recently identified protein module that has been characterized as binding to phosphopeptides containing an NPXpY motif (X = any amino acid). We describe here a novel peptide sequence recognized by the PTB domain from Drosophila Numb (dNumb), a protein involved in cell fate determination and asymmetric cell division during the development of the Drosophila nervous system. Using a Tyr-oriented peptide library to screen for ligands, the dNumb PTB domain was found to bind selectively to peptides containing a YIGPYphi motif (phi represents a hydrophobic residue). A synthetic peptide containing this sequence bound specifically to the isolated dNumb PTB domain in solution with a dissociation constant (Kd) of 5.78 +/- 0.74 microM. Interestingly, the affinity of this peptide for the dNumb PTB domain was increased (Kd = 1.41 +/- 0.10 microM) when the second tyrosine in the sequence was phosphorylated. Amino acid substitution studies of the phosphopeptide demonstrated that a core motif of sequence GP(p)Y is required for high-affinity binding to the dNumb PTB domain. Nuclear magnetic resonance experiments performed on isotopically labeled protein complexed with either Tyr- or pTyr-containing peptides suggest that the same set of amino acids in the dNumb PTB domain is involved in binding both phosphorylated and nonphosphorylated forms of the peptide. The in vitro selectivity of the dNumb PTB domain is therefore markedly different from those of the Shc and IRS-1 PTB domains, in that it interacts preferentially with a GP(p)Y motif, rather than NPXpY, and does not absolutely require ligand phosphorylation for binding. Our results suggest that the PTB domain is a versatile protein module, capable of exhibiting varied binding specificities. PMID- 9207071 TI - The regulators of G protein signaling (RGS) domains of RGS4, RGS10, and GAIP retain GTPase activating protein activity in vitro. AB - Regulators of G protein signaling (RGS) proteins accelerate GTP hydrolysis by Gi but not by Gs class alpha-subunits. All RGS proteins share a conserved 120-amino acid sequence termed the RGS domain. We have demonstrated that the RGS domains of RGS4, RGS10, and GAIP retain GTPase accelerating activity with the Gi class substrates Gialpha1, Goalpha, and Gzalpha in vitro. No regulatory activity of the RGS domains was detected for Gsalpha. Short deletions within the RGS domain of RGS4 destroyed GTPase activating protein activity and Gialpha1 substrate binding. Comparable protein-protein interactions between Gialpha1-GDP-AlF4- and the RGS domain or full-length RGS4 were detected using surface plasmon resonance. PMID- 9207070 TI - Posttranscriptional modification of retroviral primers is required for late stages of DNA replication. AB - During reverse transcription of retroviral RNA, synthesis of (-) strand DNA is primed by a cellular tRNA that anneals to an 18-nt primer binding site within the 5' long terminal repeat. For (+) strand synthesis using a (-) strand DNA template linked to the tRNA primer, only the first 18 nt of tRNA are replicated to regenerate the primer binding site, creating the (+) strand strong stop DNA intermediate and providing a 3' terminus capable of strand transfer and further elongation. On model HIV templates that approximate the (-) strand linked to natural modified or synthetic unmodified tRNA3Lys, we find that a (+) strand strong stop intermediate of the proper length is generated only on templates containing the natural, modified tRNA3Lys, suggesting that a posttranscriptional modification provides the termination signal. In the presence of a recipient template, synthesis after strand transfer occurs only from intermediates generated from templates containing modified tRNA3Lys. Reverse transcriptase from Moloney murine leukemia virus and avian myoblastosis virus shows the same requirement for a modified tRNA3Lys template. Because all retroviral tRNA primers contain the same 1-methyl-A58 modification, our results suggest that 1-methyl-A58 is generally required for termination of replication 18 nt into the tRNA sequence, generating the (+) strand intermediate, strand transfer, and subsequent synthesis of the entire (+) strand. The possibility that the host methyl transferase responsible for methylating A58 may provide a target for HIV chemotherapy is discussed. PMID- 9207072 TI - Slow dimer dissociation of the TATA binding protein dictates the kinetics of DNA binding. AB - The association of the TATA binding protein (TBP) to eukaryotic promoters is a possible rate-limiting step in gene expression. Slow promoter binding might be related to TBP's ability to occlude its DNA binding domain through dimerization. Using a "pull-down" based assay, we find that TBP dimers dissociate slowly (t1/2 = 6-10 min), and thus present a formidable kinetic barrier to TATA binding. At 10 nM, TBP appears to exist as a mixed population of monomers and dimers. In this state, TATA binding displays burst kinetics that appears to reflect rapid binding of monomers and slow dissociation of dimers. The kinetics of the slow phase is in excellent agreement with direct measurements of the kinetics of dimer dissociation. PMID- 9207073 TI - The interferon-inducible murine p48 (ISGF3gamma) gene is regulated by protooncogene c-myc. AB - p48 protein is an integral component of the multimeric interferon (IFN)-regulated transcription factor, ISGF3. We have shown earlier that this gene is regulated by a novel IFN-gamma-regulated element. In addition to the IFN-regulated element, a myc-max binding site is also present in this promoter. In this investigation we have studied the role of this site in the regulation of the p48 gene. In serum induced quiescent cells Myc up-regulated the expression of p48 mRNA. We show that the protooncogene Myc regulates the expression of p48 through the element CACGTG. Mutations in this motif abolish Myc-inducibility of the reporter genes carrying p48 promoter elements. Purified Myc and Max proteins interact with the Myc stimulated element of the p48 promoter. We also show that cells lacking p48 expression are highly susceptible to the cytocidal action of anticancer drugs. Taken together these data suggest that p48 may function as an anti-stress cell survival factor. PMID- 9207074 TI - Gene cloning, sequence analysis, and expression of 2-methyl-3-hydroxypyridine-5 carboxylic acid oxygenase. AB - The gene encoding 2-methyl-3-hydroxypyridine-5-carboxylic acid oxygenase (MHPCO; EC 1.14.12.4) was cloned by using an oligonucleotide probe corresponding to the N terminus of the enzyme to screen a DNA library of Pseudomonas sp. MA-1. The gene encodes for a protein of 379 amino acid residues corresponding to a molecular mass of 41.7 kDa, the same as that previously estimated for MHPCO. MHPCO was expressed in Escherichia coli and found to have the same properties as the native enzyme from Pseudomonas sp. MA-1. This study shows that MHPCO is a homotetrameric protein with one flavin adenine dinucleotide bound per subunit. Sequence comparison of the enzyme with other hydroxylases reveals regions that are conserved among aromatic flavoprotein hydroxylases. PMID- 9207075 TI - Requirement of STAT5b for sexual dimorphism of body growth rates and liver gene expression. AB - The signal transducer and activator of transcription, STAT5b, has been implicated in signal transduction pathways for a number of cytokines and growth factors, including growth hormone (GH). Pulsatile but not continuous GH exposure activates liver STAT5b by tyrosine phosphorylation, leading to dimerization, nuclear translocation, and transcriptional activation of the STAT, which is proposed to play a key role in regulating the sexual dimorphism of liver gene expression induced by pulsatile plasma GH. We have evaluated the importance of STAT5b for the physiological effects of GH pulses using a mouse gene knockout model. STAT5b gene disruption led to a major loss of multiple, sexually differentiated responses associated with the sexually dimorphic pattern of pituitary GH secretion. Male-characteristic body growth rates and male-specific liver gene expression were decreased to wild-type female levels in STAT5b-/- males, while female-predominant liver gene products were increased to a level intermediate between wild-type male and female levels. Although these responses are similar to those observed in GH-deficient Little mice, STAT5b-/- mice are not GH-deficient, suggesting that they may be GH pulse-resistant. Indeed, the dwarfism, elevated plasma GH, low plasma insulin-like growth factor I, and development of obesity seen in STAT5b-/- mice are all characteristics of Laron-type dwarfism, a human GH resistance disease generally associated with a defective GH receptor. The requirement of STAT5b to maintain sexual dimorphism of body growth rates and liver gene expression suggests that STAT5b may be the major, if not the sole, STAT protein that mediates the sexually dimorphic effects of GH pulses in liver and perhaps other target tissues. STAT5b thus has unique physiological functions for which, surprisingly, the highly homologous STAT5a is unable to substitute. PMID- 9207076 TI - Distinct roles for E2F proteins in cell growth control and apoptosis. AB - E2F transcription activity is composed of a family of heterodimers encoded by distinct genes. Through the overproduction of each of the five known E2F proteins in mammalian cells, we demonstrate that a large number of genes encoding proteins important for cell cycle regulation and DNA replication can be activated by the E2F proteins and that there are distinct specificities in the activation of these genes by individual E2F family members. Coexpression of each E2F protein with the DP1 heterodimeric partner does not significantly alter this specificity. We also find that only E2F1 overexpression induces cells to undergo apoptosis, despite the fact that at least two other E2F family members, E2F2 and E2F3, are equally capable of inducing S phase. The ability of E2F1 to induce apoptosis appears to result from the specific induction of an apoptosis-promoting activity rather than the lack of induction of a survival activity, because co-expression of E2F2 and E2F3 does not rescue cells from E2F1-mediated apoptosis. We conclude that E2F family members play distinct roles in cell cycle control and that E2F1 may function as a specific signal for the initiation of an apoptosis pathway that must normally be blocked for a productive proliferation event. PMID- 9207077 TI - Multiprotein bridging factor 1 (MBF1) is an evolutionarily conserved transcriptional coactivator that connects a regulatory factor and TATA element binding protein. AB - Multiprotein bridging factor 1 (MBF1) is a transcriptional cofactor that bridges between the TATA box-binding protein (TBP) and the Drosophila melanogaster nuclear hormone receptor FTZ-F1 or its silkworm counterpart BmFTZ-F1. A cDNA clone encoding MBF1 was isolated from the silkworm Bombyx mori whose sequence predicts a basic protein consisting of 146 amino acids. Bacterially expressed recombinant MBF1 is functional in interactions with TBP and a positive cofactor MBF2. The recombinant MBF1 also makes a direct contact with FTZ-F1 through the C terminal region of the FTZ-F1 DNA-binding domain and stimulates the FTZ-F1 binding to its recognition site. The central region of MBF1 (residues 35-113) is essential for the binding of FTZ-F1, MBF2, and TBP. When the recombinant MBF1 was added to a HeLa cell nuclear extract in the presence of MBF2 and FTZ622 bearing the FTZ-F1 DNA-binding domain, it supported selective transcriptional activation of the fushi tarazu gene as natural MBF1 did. Mutations disrupting the binding of FTZ622 to DNA or MBF1, or a MBF2 mutation disrupting the binding to MBF1, all abolished the selective activation of transcription. These results suggest that tethering of the positive cofactor MBF2 to a FTZ-F1-binding site through FTZ-F1 and MBF1 is essential for the binding site-dependent activation of transcription. A homology search in the databases revealed that the deduced amino acid sequence of MBF1 is conserved across species from yeast to human. PMID- 9207079 TI - A model of excitation and adaptation in bacterial chemotaxis. AB - Bacterial chemotaxis is widely studied because of its accessibility and because it incorporates processes that are important in a number of sensory systems: signal transduction, excitation, adaptation, and a change in behavior, all in response to stimuli. Quantitative data on the change in behavior are available for this system, and the major biochemical steps in the signal transduction/processing pathway have been identified. We have incorporated recent biochemical data into a mathematical model that can reproduce many of the major features of the intracellular response, including the change in the level of chemotactic proteins to step and ramp stimuli such as those used in experimental protocols. The interaction of the chemotactic proteins with the motor is not modeled, but we can estimate the degree of cooperativity needed to produce the observed gain under the assumption that the chemotactic proteins interact directly with the motor proteins. PMID- 9207078 TI - Distal enhancer regulation by promoter derepression in topologically constrained DNA in vitro. AB - Long-range promoter-enhancer interactions are a crucial regulatory feature of many eukaryotic genes yet little is known about the mechanisms involved. Using cloned chicken betaA-globin genes, either individually or within the natural chromosomal locus, enhancer-dependent transcription is achieved in vitro at a distance of 2 kb with developmentally staged erythroid extracts. This occurs by promoter derepression and is critically dependent upon DNA topology. In the presence of the enhancer, genes must exist in a supercoiled conformation to be actively transcribed, whereas relaxed or linear templates are inactive. Distal protein-protein interactions in vitro may be favored on supercoiled DNA because of topological constraints. In this system, enhancers act primarily to increase the probability of rapid and efficient transcription complex formation and initiation. Repressor and activator proteins binding within the promoter, including erythroid-specific GATA-1, mediate this process. PMID- 9207080 TI - HLA class I and II antigens are partially co-clustered in the plasma membrane of human lymphoblastoid cells. AB - Major histocompatibility complex (MHC) class II molecules displayed clustered patterns at the surfaces of T (HUT-102B2) and B (JY) lymphoma cells characterized by interreceptor distances in the micrometer range as detected by scanning force microscopy of immunogold-labeled antigens. Electron microscopy revealed that a fraction of the MHC class II molecules was also heteroclustered with MHC class I antigens at the same hierarchical level as described by the scanning force microscopy data, after specifically and sequentially labeling the antigens with 30- and 15-nm immunogold beads. On JY cells the estimated fraction of co clustered HLA II was 0.61, whereas that of the HLA I was 0.24. Clusterization of the antigens was detected by the deviation of their spatial distribution from the Poissonian distribution representing the random case. Fluorescence resonance energy transfer measurements also confirmed partial co-clustering of the HLA class I and II molecules at another hierarchical level characterized by the 2- to 10-nm Forster distance range and providing fine details of the molecular organization of receptors. The larger-scale topological organization of the MHC class I and II antigens may reflect underlying membrane lipid domains and may fulfill significant functions in cell-to-cell contacts and signal transduction. PMID- 9207081 TI - Movements of truncated kinesin fragments with a short or an artificial flexible neck. AB - To investigate the role of the neck domain of kinesin, we used optical trapping nanometry to perform high-resolution measurements of the movements and forces produced by recombinant kinesin fragments in which the neck domains were shortened or replaced by an artificial random coil. Truncated kinesin fragments (K351) that contain a motor domain consisting of approximately 340 aa and a short neck domain consisting of approximately 11 aa showed fast movement (800 nm/s) and 8-nm steps. Such behavior was similar to that of recombinant fragments containing the full-length neck domain (K411) and to that of native kinesin. Kinesin fragments lacking the short neck domain (K340), however, showed very slow movement (<50 nm/s), as previously reported. Joining an artificial 11-aa sequence that was expected to form a flexible random chain to the motor domain (K340 chain) produced normal fast ( approximately 700 nm/s) and stepwise movement. The results suggest that the neck domain does not act as a rigid lever arm to magnify the structural change at the catalytic domain as has been believed for myosin, but it does act as a flexible joint to guarantee the mobility of the motor domain. PMID- 9207082 TI - Prion protein NMR structure and species barrier for prion diseases. AB - The structural basis of species specificity of transmissible spongiform encephalopathies, such as bovine spongiform encephalopathy or "mad cow disease" and Creutzfeldt-Jakob disease in humans, has been investigated using the refined NMR structure of the C-terminal domain of the mouse prion protein with residues 121-231. A database search for mammalian prion proteins yielded 23 different sequences for the fragment 124-226, which display a high degree of sequence identity and show relevant amino acid substitutions in only 18 of the 103 positions. Except for a unique isolated negative surface charge in the bovine protein, the amino acid differences are clustered in three distinct regions of the three-dimensional structure of the cellular form of the prion protein. Two of these regions represent potential species-dependent surface recognition sites for protein-protein interactions, which have independently been implicated from in vitro and in vivo studies of prion protein transformation. The third region consists of a cluster of interior hydrophobic side chains that may affect prion protein transformation at later stages, after initial conformational changes in the cellular protein. PMID- 9207083 TI - Nuclear punctate distribution of ALL-1 is conferred by distinct elements at the N terminus of the protein. AB - The ALL-1 gene positioned at 11q23 is directly involved in human acute leukemia either through a variety of chromosome translocations or by partial tandem duplications. ALL-1 is the human homologue of Drosophila trithorax which plays a critical role in maintaining proper spatial and temporal expression of the Antennapedia-bithorax homeotic genes determining the fruit fly's body pattern. Utilizing specific antibodies, we found that the ALL-1 protein distributes in cultured cells in a nuclear punctate pattern. Several chimeric ALL-1 proteins encoded by products of the chromosome translocations and expressed in transfected cells showed similar speckles. Dissection of the ALL-1 protein identified within its approximately 1,100 N-terminal residues three polypeptides directing nuclear localization and at least two main domains conferring distribution in dots. The latter spanned two short sequences conserved with TRITHORAX. Enforced nuclear expression of other domains of ALL-1, such as the PHD (zinc) fingers and the SET motif, resulted in uniform nonpunctate patterns. This indicates that positioning of the ALL-1 protein in subnuclear structures is mediated via interactions of ALL 1 N-terminal elements. We suggest that the speckles represent protein complexes which contain multiple copies of the ALL-1 protein and are positioned at ALL-1 target sites on the chromatin. Therefore, the role of the N-terminal portion of ALL-1 is to direct the protein to its target genes. PMID- 9207084 TI - Fas-induced apoptosis of T cells occurs independently of ceramide generation. AB - The Fas receptor is one of a number of important physiological inducers of programmed cell death (apoptosis). Current models for regulation of this process involve rapid conversion of sphingomyelin to ceramide by cellular sphingomyelinases. Induced changes in cellular levels of such sphingosine-based ceramides are normally extrapolated from measurements of sphingomyelinase activity or following their conversion to ceramide phosphate by treatment of cellular lipid extracts with bacterial diacylglycerol kinase (DAGK). To allow direct study of cellular sphingosine- and sphinganine-based ceramide levels, we developed a mass spectrometric technique capable of determining inducible changes in both overall ceramide levels and species distribution in cellular lipid preparations. Contrary to current models, we detected no changes in cellular ceramide levels up to 2 hr poststimulation of Jurkat T cells with an anti-Fas IgM, although this treatment did induce apoptosis. We also determined in the same system that, when utilizing the DAGK assay, increased phosphorylation of substrates that comigrated with ceramide standards was apparent but that this effect was due to an enhancement of DAGK activity rather than increases in levels of cellular ceramides as substrates per se. Thus, the first direct measurement of ceramides present in cells undergoing apoptosis indicates that, insofar as it can be measured, the induction of apoptosis does not involve the generation of sphingosine-based ceramides, contrary to many published accounts. PMID- 9207086 TI - Requirement of poly(ADP-ribose) polymerase in recovery from DNA damage in mice and in cells. AB - Poly(ADP-ribose) polymerase [PARP; NAD+ ADP-ribosyltransferase; NAD+: poly(adenosine-diphosphate-D-ribosyl)-acceptor ADP-D-ribosyltransferase, EC 2.4.2.30] is a zinc-finger DNA-binding protein that detects specifically DNA strand breaks generated by genotoxic agents. To determine its biological function, we have inactivated both alleles by gene targeting in mice. Treatment of PARP-/- mice either by the alkylating agent N-methyl-N-nitrosourea (MNU) or by gamma-irradiation revealed an extreme sensitivity and a high genomic instability to both agents. Following whole body gamma-irradiation (8 Gy) mutant mice died rapidly from acute radiation toxicity to the small intestine. Mice-derived PARP-/ cells displayed a high sensitivity to MNU exposure: a G2/M arrest in mouse embryonic fibroblasts and a rapid apoptotic response and a p53 accumulation were observed in splenocytes. Altogether these results demonstrate that PARP is a survival factor playing an essential and positive role during DNA damage recovery. PMID- 9207085 TI - Structural protein 4.1 is located in mammalian centrosomes. AB - Structural protein 4.1 was first characterized as an important 80-kDa protein in the mature red cell membrane skeleton. It is now known to be a member of a family of protein isoforms detected at diverse intracellular sites in many nucleated mammalian cells. We recently reported that protein 4.1 isoforms are present at interphase in nuclear matrix and are rearranged during the cell cycle. Here we report that protein 4.1 epitopes are present in centrosomes of human and murine cells and are detected by using affinity-purified antibodies specific for 80-kDa red cell 4.1 and for 4.1 peptides. Immunofluorescence, by both conventional and confocal microscopy, showed that protein 4.1 epitopes localized in the pericentriolar region. Protein 4.1 epitopes remained in centrosomes after extraction of cells with detergent, salt, and DNase. Higher resolution electron microscopy of detergent-extracted cell whole mounts showed centrosomal protein 4.1 epitopes distributed along centriolar cylinders and on pericentriolar fibers, at least some of which constitute the filamentous network surrounding each centriole. Double-label electron microscopy showed that protein 4.1 epitopes were predominately localized in regions also occupied by epitopes for centrosome specific autoimmune serum 5051 but were not found on microtubules. Our results suggest that protein 4.1 is an integral component of centrosome structure, in which it may play an important role in centrosome function during cell division and organization of cellular architecture. PMID- 9207087 TI - The cluA- mutant of Dictyostelium identifies a novel class of proteins required for dispersion of mitochondria. AB - The cluA gene of Dictyostelium discoideum encodes a novel 150-kDa protein. Disruption of cluA results in clustering of mitochondria near the cell center. This is a striking difference from normal cells, whose mitochondria are dispersed uniformly throughout the cytoplasm. The mutant cell populations also exhibit an increased frequency of multinucleated cells, suggesting an impairment in cytokinesis. Both phenotypes are reversed by transformation of cluA- cells with a plasmid carrying a constitutively expressed cluA gene. The predicted sequence of the cluA gene product is homologous to sequences encoded by open reading frames in the genomes of Saccharomyces cerevisiae and Caenorhabditis elegans, but not to any known protein. The only exception is a short region with some homology to the 42-residue imperfect repeats present in the kinesin light chain, which probably function in protein-protein interaction. These studies identify a new class of proteins that appear to be required for the proper distribution of mitochondria. PMID- 9207088 TI - In vivo zippering of inner and outer mitochondrial membranes by a stable translocation intermediate. AB - It was previously assumed that the import of cytoplasmically synthesized precursor proteins into mitochondria occurs through a single structure spanning both outer and inner membranes at contact sites. Based on recent findings, however, the two membranes appear to contain independent translocation elements that reversibly cooperate during protein import. This feature makes it difficult to generate a means of isolating a fully integrated and functional translocation complex. To study these independent translocases in vitro and in vivo, we have constructed a chimeric protein consisting of an N-terminal authentic mitochondrial precursor (delta1-pyrroline-5-carboxylate dehydrogenase) linked, through glutathione S-transferase, to IgG binding domains derived from staphylococcal protein A. This construct becomes trapped en route to the matrix, spanning both outer and inner membranes in such a way that the entire signal-less delta1-pyrroline-5-carboxylate dehydrogenase moiety reaches the matrix, while only the folded protein A domain remains outside. During in vivo import of this precursor, outer and inner membranes of yeast mitochondria become progressively "zippered" together, forming long stretches of close contact. Using this novel intermediate, the outer and inner mitochondrial membrane channels, which normally interact only transiently, can be tightly joined (both in vitro and in vivo), forming a stable association. This suggests a method for isolating the functional translocation complex as a single entity. PMID- 9207089 TI - Ethanolamine modulates the rate of rat hepatocyte proliferation in vitro and in vivo. AB - A low molecular weight, heat-resistant hepatotrophic factor in an extract from the bovine intestinal mucosa was purified and identified as ethanolamine by structural analyses. The mode of action of ethanolamine in vitro and in vivo coincided with that of the crude extract of the tissue, indicating that ethanolamine is the active component. Ethanolamine synergistically elevated the stimulation of DNA synthesis in hepatocytes in primary culture when added together with a growth factor, such as epidermal growth factor, with the ED50 being 20 microM, although it showed little stimulatory effect by itself. Contrary to these in vitro results, the intraperitoneal administration of ethanolamine hydrochloride (24 mg of ethanolamine per kg of body weight) enhanced hepatocyte proliferation in regenerating rat livers after two-thirds hepatectomy without the administration of any growth factors. In the regenerating liver, hepatocyte proliferation may be initiated by an endogenous growth factor, but the supply of ethanolamine in circulation may not be sufficient for optimal hepatocyte proliferation; thus, the exogenous administration of ethanolamine may further enhance hepatocyte proliferation. Ethanolamine in circulation may be a humoral hepatotrophic factor. PMID- 9207090 TI - Identification of an early endosomal protein regulated by phosphatidylinositol 3 kinase. AB - Phosphatidylinositol 3-kinases (PI 3-kinases) have been implicated in membrane trafficking in the secretory and endocytic pathways of yeast and mammalian cells, but the molecular mechanisms by which these lipid kinases operate are not known. Here we identify a protein of 170 kDa that is rapidly released from cell membranes in response to wortmannin, a potent inhibitor of mammalian PI 3 kinases. The amino acid sequence of peptides from p170 reveal its identity to early endosomal antigen (EEA) 1, an endosomal antigen with homology to several yeast proteins genetically implicated in membrane trafficking. Immunofluorescence analysis of 3T3-L1 adipocytes with antisera against p170/EEA1 reveal a punctate peripheral pattern that becomes diffuse in response to wortmannin. In vitro, p170/EEA1 binds specifically to liposomes containing PIns(3)P, suggesting that the effect of wortmannin on cells is due to inhibition of PIns(3)P production. Thus, p170/EEA1 may define a family of proteins that mediate the regulatory effects of 3'-phosphoinositides on membrane trafficking in yeast and mammalian cells. PMID- 9207091 TI - Mso1p: a yeast protein that functions in secretion and interacts physically and genetically with Sec1p. AB - The yeast Sec1p protein functions in the docking of secretory transport vesicles to the plasma membrane. We previously have cloned two yeast genes encoding syntaxins, SSO1 and SSO2, as suppressors of the temperature-sensitive sec1-1 mutation. We now describe a third suppressor of sec1-1, which we call MSO1. Unlike SSO1 and SSO2, MSO1 is specific for sec1 and does not suppress mutations in any other SEC genes. MSO1 encodes a small hydrophilic protein that is enriched in a microsomal membrane fraction. Cells that lack MSO1 are viable, but they accumulate secretory vesicles in the bud, indicating that the terminal step in secretion is partially impaired. Moreover, loss of MSO1 shows synthetic lethality with mutations in SEC1, SEC2, and SEC4, and other synthetic phenotypes with mutations in several other late-acting SEC genes. We further found that Mso1p interacts with Sec1p both in vitro and in the two-hybrid system. These findings suggest that Mso1p is a component of the secretory vesicle docking complex whose function is closely associated with that of Sec1p. PMID- 9207092 TI - Mitogen-activated protein kinase kinase 7 is an activator of the c-Jun NH2 terminal kinase. AB - The c-Jun NH2-terminal kinase (JNK) group of mitogen-activated protein (MAP) kinases is activated by phosphorylation on Thr and Tyr. Here we report the molecular cloning of a new member of the mammalian MAP kinase kinase group (MKK7) that functions as an activator of JNK. In vitro protein kinase assays demonstrate that MKK7 phosphorylates and activates JNK, but not the p38 or extracellular signal-regulated kinase groups of MAP kinase. Expression of MKK7 in cultured cells causes activation of the JNK signal transduction pathway. MKK7 is therefore established to be a novel component of the JNK signal transduction pathway. PMID- 9207094 TI - Suspensor-derived polyembryony caused by altered expression of valyl-tRNA synthetase in the twn2 mutant of Arabidopsis. AB - The twn2 mutant of Arabidopsis exhibits a defect in early embryogenesis where, following one or two divisions of the zygote, the decendents of the apical cell arrest. The basal cells that normally give rise to the suspensor proliferate abnormally, giving rise to multiple embryos. A high proportion of the seeds fail to develop viable embryos, and those that do, contain a high proportion of partially or completely duplicated embryos. The adult plants are smaller and less vigorous than the wild type and have a severely stunted root. The twn2-1 mutation, which is the only known allele, was caused by a T-DNA insertion in the 5' untranslated region of a putative valyl-tRNA synthetase gene, valRS. The insertion causes reduced transcription of the valRS gene in reproductive tissues and developing seeds but increased expression in leaves. Analysis of transcript initiation sites and the expression of promoter-reporter fusions in transgenic plants indicated that enhancer elements inside the first two introns interact with the border of the T-DNA to cause the altered pattern of expression of the valRS gene in the twn2 mutant. The phenotypic consequences of this unique mutation are interpreted in the context of a model, suggested by Vernon and Meinke [Vernon, D. M. & Meinke, D. W. (1994) Dev. Biol. 165, 566-573], in which the apical cell and its decendents normally suppress the embryogenic potential of the basal cell and its decendents during early embryo development. PMID- 9207093 TI - Distinct mechanisms of splicing regulation in vivo by the Drosophila protein Sex lethal. AB - The protein Sex-lethal (SXL) controls pre-mRNA splicing of two genes involved in Drosophila sex determination: transformer (tra) and the Sxl gene itself. Previous in vitro results indicated that SXL antagonizes the general splicing factor U2AF65 to regulate splicing of tra. In this report, we have used transgenic flies expressing chimeric proteins between SXL and the effector domain of U2AF65 to study the mechanisms of splicing regulation by SXL in vivo. Conferring U2AF activity to SXL relieves its inhibitory activity on tra splicing but not on Sxl splicing. Therefore, antagonizing U2AF65 can explain tra splicing regulation both in vitro and in vivo, but this mechanism cannot explain splicing regulation of Sxl pre-mRNA. These results are a direct proof that Sxl, the master regulatory gene in sex determination, has multiple and separable activities in the regulation of pre-mRNA splicing. PMID- 9207095 TI - Differentiation of pancreatic epithelial progenitor cells into hepatocytes following transplantation into rat liver. AB - The ability to identify, isolate, and transplant progenitor cells from solid tissues would greatly facilitate the treatment of diseases currently requiring whole organ transplantation. In this study, cell fractions enriched in candidate epithelial progenitor cells from the rat pancreas were isolated and transplanted into the liver of an inbred strain of Fischer rats. Using a dipeptidyl dipeptidase IV genetic marker system to follow the fate of transplanted cells in conjunction with albumin gene expression, we provide conclusive evidence that, after transplantation to the liver, epithelial progenitor cells from the pancreas differentiate into hepatocytes, express liver-specific proteins, and become fully integrated into the liver parenchymal structure. These studies demonstrate the presence of multipotent progenitor cells in the adult pancreas and establish a role for the liver microenvironment in the terminal differentiation of epithelial cells of foregut origin. They further suggest that such progenitor cells might be useful in studies of organ repopulation following acute or chronic liver injury. PMID- 9207097 TI - Evolution of plastid gene rps2 in a lineage of hemiparasitic and holoparasitic plants: many losses of photosynthesis and complex patterns of rate variation. AB - The plastid genomes of some nonphotosynthetic parasitic plants have experienced an extreme reduction in gene content and an increase in evolutionary rate of remaining genes. Nothing is known of the dynamics of these events or whether either is a direct outcome of the loss of photosynthesis. The parasitic Scrophulariaceae and Orobanchaceae, representing a continuum of heterotrophic ability ranging from photosynthetic hemiparasites to nonphotosynthetic holoparasites, are used to investigate these issues. We present a phylogenetic hypothesis for parasitic Scrophulariaceae and Orobanchaceae based on sequences of the plastid gene rps2, encoding the S2 subunit of the plastid ribosome. Parasitic Scrophulariaceae and Orobanchaceae form a monophyletic group in which parasitism can be inferred to have evolved once. Holoparasitism has evolved independently at least five times, with certain holoparasitic lineages representing single species, genera, and collections of nonphotosynthetic genera. Evolutionary loss of the photosynthetic gene rbcL is limited to a subset of holoparasitic lineages, with several holoparasites retaining a full length rbcL sequence. In contrast, the translational gene rps2 is retained in all plants investigated but has experienced rate accelerations in several hemi- as well as holoparasitic lineages, suggesting that there may be substantial molecular evolutionary changes to the plastid genome of parasites before the loss of photosynthesis. Independent patterns of synonymous and nonsynonymous rate acceleration in rps2 point to distinct mechanisms underlying rate variation in different lineages. Parasitic Scrophulariaceae (including the traditional Orobanchaceae) provide a rich platform for the investigation of molecular evolutionary process, gene function, and the evolution of parasitism. PMID- 9207098 TI - Evolution of foraging behavior in Drosophila by density-dependent selection. AB - One of the rare examples of a single major gene underlying a naturally occurring behavioral polymorphism is the foraging locus of Drosophila melanogaster. Larvae with the rover allele, forR, have significantly longer foraging path lengths on a yeast paste than do those homozygous for the sitter allele, fors. These variants do not differ in general activity in the absence of food. The evolutionary significance of this polymorphism is not as yet understood. Here we examine the effect of high and low animal rearing densities on the larval foraging path length phenotype and show that density-dependent natural selection produces changes in this trait. In three unrelated base populations the long path (rover) phenotype was selected for under high-density rearing conditions, whereas the short path (sitter) phenotype was selected for under low-density conditions. Genetic crosses suggested that these changes resulted from alterations in the frequency of the fors allele in the low-density-selected lines. Further experiments showed that density-dependent selection during the larval stage rather than the adult stage of development was sufficient to explain these results. Density-dependent mechanisms may be sufficient to maintain variation in rover and sitter behavior in laboratory populations. PMID- 9207099 TI - Substantial narrowing of the Niemann-Pick C candidate interval by yeast artificial chromosome complementation. AB - Niemann-Pick disease type C (NP-C) is an autosomal recessive lipidosis linked to chromosome 18q11-12, characterized by lysosomal accumulation of unesterified cholesterol and delayed induction of cholesterol-mediated homeostatic responses. This cellular phenotype is identifiable cytologically by filipin staining and biochemically by measurement of low-density lipoprotein-derived cholesterol esterification. The mutant Chinese hamster ovary cell line (CT60), which displays the NP-C cellular phenotype, was used as the recipient for a complementation assay after somatic cell fusions with normal and NP-C murine cells suggested that this Chinese hamster ovary cell line carries an alteration(s) in the hamster homolog(s) of NP-C. To narrow rapidly the candidate interval for NP-C, three overlapping yeast artificial chromosomes (YACs) spanning the 1 centimorgan human NP-C interval were introduced stably into CT60 cells and analyzed for correction of the cellular phenotype. Only YAC 911D5 complemented the NP-C phenotype, as evidenced by cytological and biochemical analyses, whereas no complementation was obtained from the other two YACs within the interval or from a YAC derived from chromosome 7. Fluorescent in situ hybridization indicated that YAC 911D5 was integrated at a single site per CT60 genome. These data substantially narrow the NP-C critical interval and should greatly simplify the identification of the gene responsible in mouse and man. This is the first demonstration of YAC complementation as a valuable adjunct strategy for positional cloning of a human gene. PMID- 9207100 TI - Direct isolation of human BRCA2 gene by transformation-associated recombination in yeast. AB - Mutant forms of the BRCA2 gene contribute significantly to hereditary breast cancer. Isolation of the normal and mutant forms of the BRCA2 gene with its natural promoter would greatly facilitate analysis of the gene and its contribution to breast cancer. We have accomplished the direct isolation of the 90-kb gene from total human DNA by transformation-associated recombination in yeast using a small amount of 5' and 3' BRCA2 sequence information. Because the entire isolation procedure of a single chromosomal gene could be accomplished in approximately 2 weeks, the transformation-associated recombination cloning approach is readily applicable to studies of chromosome alterations and human genetic diseases. PMID- 9207101 TI - Expansion of a CUG trinucleotide repeat in the 3' untranslated region of myotonic dystrophy protein kinase transcripts results in nuclear retention of transcripts. AB - Expansion of a CTG trinucleotide repeat in the 3' untranslated region (UTR) of DMPK, the gene encoding myotonic dystrophy protein kinase, induces the dominantly inherited neuromuscular disorder myotonic dystrophy (DM). Transcripts containing the expanded trinucleotide are abundant in differentiated cultured myoblasts, and they are spliced and polyadenylylated normally. However, mutant transcripts never reach the cytoplasm in these nonmitotic cells; instead, they form stable clusters that are tightly linked to the nuclear matrix, which can prevent effective biochemical purification of these transcripts. In DM patients, reduced DMPK protein levels, consequent to nuclear retention of mutant transcripts, are probably a cause of disease development. Formation of nuclear foci is a novel mechanism for preventing transcript export and effecting a loss of gene function. PMID- 9207102 TI - Transcriptional abnormality in myotonic dystrophy affects DMPK but not neighboring genes. AB - Myotonic dystrophy (DM) is caused by the expansion of a trinucleotide repeat, CTG, in the 3' untranslated region of a protein kinase gene, DMPK. We set out to determine what effect this expanded repeat has on RNA processing. The subcellular fractionation of RNA and the separate analysis of DMPK transcripts from each allele reveals that transcripts from expanded DMPK alleles are retained within the nucleus and are absent from the cytoplasm of DM cell lines. The nuclear retention of DMPK transcripts occurs above a critical threshold between 80 and 400 CTGs. Further analysis of the nuclear RNA reveals an apparent reduction in the proportion of expansion-derived DMPK transcripts after poly(A)+ selection. Quantitative analysis of RNA also indicates that although the level of cytoplasmic DMPK transcript is altered in DM patients, the levels of transcripts from 59 and DMAHP, two genes that immediately flank DMPK, are unaffected in DM cell lines. PMID- 9207103 TI - Tissue-specific knockout of the mouse Pig-a gene reveals important roles for GPI anchored proteins in skin development. AB - Glycosylphosphatidylinositol (GPI)-anchored proteins are widely distributed on plasma membranes of eukaryotes. More than 50 GPI-anchored proteins have been shown to be spatiotemporally expressed in mice with a deficiency of GPI-anchor biosynthesis that causes embryonic lethality. Here, we examine the functional roles of GPI-anchored proteins in mouse skin using the Cre-loxP recombination system. We disrupted the Pig-a gene, an X-linked gene essential for GPI-anchor biosynthesis, in skin. The Cre-mediated Pig-a disruption occurred in skin at almost 100% efficiency in male mice bearing two identically orientated loxP sites within the Pig-a gene. Expression of GPI-anchored proteins was completely absent in the skin of these mice. The skin of such mutants looked wrinkled and more scaly than that of wild-type mice. Furthermore, histological examination of mutant mice showed that the epidermal horny layer was tightly packed and thickened. Electron microscopy showed that the intercellular space was narrow and there were many small vesicles embedded in the intercellular space that were not observed in equivalent wild-type mouse skin preparations. Mutant mice died within a few days after birth, suggesting that Pig-a function is essential for proper skin differentiation and maintenance. PMID- 9207104 TI - Cloning of a gene (RIG-G) associated with retinoic acid-induced differentiation of acute promyelocytic leukemia cells and representing a new member of a family of interferon-stimulated genes. AB - In a cell line (NB4) derived from a patient with acute promyelocytic leukemia, all-trans-retinoic acid (ATRA) and interferon (IFN) induce the expression of a novel gene we call RIG-G (for retinoic acid-induced gene G). This gene codes for a 58-kDa protein containing 490 amino acids with several potential sites for post translational modification. In untreated NB4 cells, the expression of RIG-G is undetectable. ATRA treatment induces the transcriptional expression of RIG-G relatively late (12-24 hr) in a protein synthesis-dependent manner, whereas IFN alpha induces its expression early (30 min to 3 hr). Database search has revealed a high-level homology between RIG-G and several IFN-stimulated genes in human (ISG54K, ISG56K, and IFN-inducible and retinoic acid-inducible 58K gene) and some other species, defining a well conserved gene family. The gene is composed of two exons and has been mapped by fluorescence in situ hybridization to chromosome 10q24, where two other human IFN-stimulated gene members are localized. A synergistic induction of RIG-G expression in NB4 cells by combined treatment with ATRA and IFNs suggests that a collaboration exists between their respective signaling pathways. PMID- 9207105 TI - Silent chromatin determines target preference of the Saccharomyces retrotransposon Ty5. AB - The HML and HMR mating loci of Saccharomyces cerevisiae are bound in silent chromatin, which is assembled at the flanking E and I transcriptional silencers. The retrotransposon Ty5 preferentially integrates into regions of silent chromatin, and Ty5 insertions near the HMR-E silencer account for approximately 2% of total transposition events. Most Ty5 insertions occur within 800 bp on either side of the autonomously replicating consensus sequence within HMR-E. Ty5 target preference is determined by silent chromatin, because integration near HMR E is abolished in strains with silencer mutations that alleviate transcriptional repression. The recognition of specific DNA sequences per se does not direct integration, rather, it is the protein complex assembled at the silencers. As demonstrated here for Ty5, recognition of specific chromatin domains may be a general mechanism by which retrotransposons and retroviruses determine integration sites. PMID- 9207106 TI - Isolation of human and mouse genes based on homology to REC2, a recombinational repair gene from the fungus Ustilago maydis. AB - A human and a mouse gene have been isolated based on homology to a recombinational repair gene from the corn smut Ustilago maydis. The new human (h) gene, termed hREC2, bears striking resemblance to several others, including hRAD51 and hLIM15. hREC2 is located on human chromosome 14 at q23-24. The overall amino acid sequence reveals characteristic elements of a RECA-like gene yet harbors an src-like phosphorylation site curiously absent from hRAD51 and hLIM15. Unlike these two relatives, hREC2 is expressed in a wide range of tissues including lung, liver, placenta, pancreas, leukocytes, colon, small intestine, brain, and heart, as well as thymus, prostate, spleen, and uterus. Of greatest interest is that hREC2 is undetectable by reverse transcription-coupled PCR in tissue culture unless the cells are treated by ionizing radiation. PMID- 9207107 TI - Telomeres in the mouse have large inter-chromosomal variations in the number of T2AG3 repeats. AB - The ultra-long telomeres that have been observed in mice are not in accordance with the concept that critical telomere shortening is related to aging and immortalization. Here, we have used quantitative fluorescence in situ hybridization to estimate (T2AG3)n lengths of individual telomeres in various mouse strains. Telomere lengths were very heterogeneous, but specific chromosomes of bone marrow cells and skin fibroblasts from individual mice had similar telomere lengths. We estimate that the shortest telomeres are around 10 kb in length, indicating that each mouse cell has a few telomeres with (T2AG3)n lengths within the range of human telomeres. These short telomeres may be critical in limiting the replicative potential of murine cells. PMID- 9207108 TI - Cloning and characterization of a mammalian 8-oxoguanine DNA glycosylase. AB - Oxidative DNA damage is generated by reactive oxygen species. The mutagenic base, 8-oxoguanine, formed by this process, is removed from oxidatively damaged DNA by base excision repair. Genes coding for DNA repair enzymes that recognize 8 oxoguanine have been reported in bacteria and yeast. We have identified and characterized mouse and human cDNAs encoding homologs of the 8-oxoguanine DNA glycosylase (ogg1) gene of Saccharomyces cerevisiae. Escherichia coli doubly mutant for mutM and mutY have a mutator phenotype and are deficient in 8 oxoguanine repair. The recombinant mouse gene (mOgg1) suppresses the mutator phenotype of mutY/mutM E. coli. Extracts prepared from mutY/mutM E. coli expressing mOgg1 contain an activity that excises 8-oxoguanine from DNA and a beta-lyase activity that nicks DNA 3' to the lesion. The mouse ogg1 gene product acts efficiently on DNA duplexes in which 7, 8-dihydroxy-8-oxo-2'-deoxyguanosine (8-oxodG) is paired with dC, acts weakly on duplexes in which 8-oxodG is paired with dT or dG, and is inactive against duplexes in which 8-oxodG is paired with dA. Mouse and human ogg1 genes contain a helix-hairpin-helix structural motif with conserved residues characteristic of a recently defined family of DNA glycosylases. Ogg1 mRNA is expressed in several mouse tissues; highest levels were detected in testes. Isolation of the mouse ogg1 gene makes it possible to modulate its expression in mice and to explore the involvement of oxidative DNA damage and associated repair processes in aging and cancer. PMID- 9207109 TI - Specific cleavage of chromosomal and plasmid DNA strands in gram-positive and gram-negative bacteria can be detected with nucleotide resolution. AB - A sensitive and precise in vitro technique for detecting DNA strand discontinuities produced in vivo has been developed. The procedure, a form of runoff DNA synthesis on molecules released from lysed bacterial cells, mapped precisely the position of cleavage of the plasmid pMV158 leading strand origin in Streptococcus pneumoniae and the site of strand scission, nic, at the transfer origins of F and the F-like plasmid R1 in Escherichia coli. When high frequency of recombination strains of E. coli were examined, DNA strand discontinuities at the nic positions of the chromosomally integrated fertility factors were also observed. Detection of DNA strand scission at the nic position of F DNA in the high frequency of recombination strains, as well as in the episomal factors, was dependent on sexual expression from the transmissable element, but was independent of mating. These results imply that not only the transfer origins of extrachromosomal F and F-like fertility factors, but also the origins of stably integrated copies of these plasmids, are subject to an equilibrium of cleavage and ligation in vivo in the absence of DNA transfer. PMID- 9207111 TI - Position- and orientation-independent activity of the Schizosaccharomyces pombe meiotic recombination hot spot M26. AB - The activity of the M26 meiotic recombination hot spot of Schizosaccharomyces pombe depends on the presence of the heptamer 5'-ATGACGT-3'. Transplacement of DNA fragments containing the ade6-M26 gene to other chromosomal loci has previously demonstrated that the heptamer functions in some, but not all, transplacements, suggesting that hot spot activity depends on chromosomal context. In this study, hot spot activity was tested in the absence of gross DNA changes by using site-directed mutagenesis to create the heptamer sequence at novel locations in the genome. When created by mutagenesis of 1-4 bp in the ade6 and ura4 genes, the heptamer was active as a recombination hot spot, in an orientation-independent manner, at all locations tested. Thus, the heptamer sequence can create an active hot spot in other chromosomal contexts, provided that the gross chromosomal structure is not altered; this result is consistent with the hypothesis that a specific higher-order chromatin structure is required for M26 hot spot activity. PMID- 9207112 TI - Evolution of the Friedreich's ataxia trinucleotide repeat expansion: founder effect and premutations. AB - Friedreich's ataxia, the most frequent inherited ataxia, is caused, in the vast majority of cases, by large GAA repeat expansions in the first intron of the frataxin gene. The normal sequence corresponds to a moderately polymorphic trinucleotide repeat with bimodal size distribution. Small normal alleles have approximately eight to nine repeats whereas a more heterogeneous mode of large normal alleles ranges from 16 to 34 GAA. The latter class accounts for approximately 17% of normal alleles. To identify the origin of the expansion mutation, we analyzed linkage disequilibrium between expansion mutations or normal alleles and a haplotype of five polymorphic markers within or close to the frataxin gene; 51% of the expansions were associated with a single haplotype, and the other expansions were associated with haplotypes that could be related to the major one by mutation at a polymorphic marker or by ancient recombination. Of interest, the major haplotype associated with expansion is also the major haplotype associated with the larger alleles in the normal size range and was almost never found associated with the smaller normal alleles. The results indicate that most if not all large normal alleles derive from a single founder chromosome and that they represent a reservoir for larger expansion events, possibly through "premutation" intermediates. Indeed, we found two such alleles (42 and 60 GAA) that underwent cataclysmic expansion to pathological range in a single generation. This stepwise evolution to large trinucleotide expansions already was suggested for myotonic dystrophy and fragile X syndrome and may relate to a common mutational mechanism, despite sequence motif differences. PMID- 9207113 TI - Synpolydactyly phenotypes correlate with size of expansions in HOXD13 polyalanine tract. AB - Synpolydactyly (SPD) is a dominantly inherited congenital limb malformation. Typical cases have 3/4 finger and 4/5 toe syndactyly, with a duplicated digit in the syndactylous web, but incomplete penetrance and variable expressivity are common. The condition has recently been shown to be caused by expansions of an imperfect trinucleotide repeat sequence encoding a 15-residue polyalanine tract in HOXD13. We have studied 16 new and 4 previously published SPD families, with between 7 and 14 extra residues in the tract, to analyze the molecular basis for the observed variation in phenotype. Although there is no evidence of change in expansion size within families, even over six generations, there is a highly significant increase in the penetrance and severity of phenotype with increasing expansion size, affecting both hands (P = 0.012) and feet (P < 0. 00005). Affected individuals from a family with a 14-alanine expansion, the largest so far reported, all have a strikingly similar and unusually severe limb phenotype, involving the first digits and distal carpals. Affected males from this family also have hypospadias, not previously described in SPD, but consistent with HOXD13 expression in the developing genital tubercle. The remarkable correlation between phenotype and expansion size suggests that expansion of the tract leads to a specific gain of function in the mutant HOXD13 protein, and has interesting implications for the role of polyalanine tracts in the control of transcription. PMID- 9207114 TI - A single amino acid substitution converts a carboxylesterase to an organophosphorus hydrolase and confers insecticide resistance on a blowfly. AB - Resistance to organophosphorus (OP) insecticides is associated with decreased carboxylesterase activity in several insect species. It has been proposed that the resistance may be the result of a mutation in a carboxylesterase that simultaneously reduces its carboxylesterase activity and confers an OP hydrolase activity (the "mutant ali-esterase hypothesis"). In the sheep blowfly, Lucilia cuprina, the association is due to a change in a specific esterase isozyme, E3, which, in resistant flies, has a null phenotype on gels stained using standard carboxylesterase substrates. Here we show that an OP-resistant allele of the gene that encodes E3 differs at five amino acid replacement sites from a previously described OP-susceptible allele. Knowledge of the structure of a related enzyme (acetylcholinesterase) suggests that one of these substitutions (Gly137 --> Asp) lies within the active site of the enzyme. The occurrence of this substitution is completely correlated with resistance across 15 isogenic strains. In vitro expression of two natural and two synthetic chimeric alleles shows that the Asp137 substitution alone is responsible for both the loss of E3's carboxylesterase activity and the acquisition of a novel OP hydrolase activity. Modeling of Asp137 in the homologous position in acetylcholinesterase suggests that Asp137 may act as a base to orientate a water molecule in the appropriate position for hydrolysis of the phosphorylated enzyme intermediate. PMID- 9207115 TI - Gene replacement by homologous recombination in the multicellular green alga Volvox carteri. AB - With only two different cell types, the haploid green alga Volvox represents the simplest multicellular model system. To facilitate genetic investigations in this organism, the occurrence of homologous recombination events was investigated with the intent of developing methods for gene replacement and gene disruption. First, homologous recombination between two plasmids was demonstrated by using overlapping nonfunctional fragments of a recombinant arylsulfatase gene (tubulin promoter/arylsulfatase gene). After bombardment of Volvox reproductive cells with DNA-coated gold microprojectiles, transformants expressing arylsulfatase constitutively were recovered, indicating the presence of the machinery for homologous recombination in Volvox. Second, a well characterized loss-of-function mutation in the nuclear nitrate reductase gene (nitA) with a single G --> A nucleotide exchange in a 5'-splice site was chosen as a target for gene replacement. Gene replacement by homologous recombination was observed with a reasonably high frequency only if the replacement vector containing parts of the functional nitrate reductase gene contained only a few nucleotide exchanges. The ratio of homologous to random integration events ranged between 1:10 and 1:50, i.e., homologous recombination occurs frequently enough in Volvox to apply the powerful tool of gene disruption for functional studies of novel genes. PMID- 9207116 TI - Three novel families of miniature inverted-repeat transposable elements are associated with genes of the yellow fever mosquito, Aedes aegypti. AB - Three novel families of transposable elements, Wukong, Wujin, and Wuneng, are described in the yellow fever mosquito, Aedes aegypti. Their copy numbers range from 2,100 to 3,000 per haploid genome. There are high degrees of sequence similarity within each family, and many structural but not sequence similarities between families. The common structural characteristics include small size, no coding potential, terminal inverted repeats, potential to form a stable secondary structure, A+T richness, and putative 2- to 4-bp A+T-biased specific target sites. Evidence of previous mobility is presented for the Wukong elements. Elements of these three families are associated with 7 of 16 fully or partially sequenced Ae. aegypti genes. Characteristics of these mosquito elements indicate strong similarities to the miniature inverted-repeat transposable elements (MITEs) recently found to be associated with plant genes. MITE-like elements have also been reported in two species of Xenopus and in Homo sapiens. This characterization of multiple families of highly repetitive MITE-like elements in an invertebrate extends the range of these elements in eukaryotic genomes. A hypothesis is presented relating genome size and organization to the presence of highly reiterated MITE families. The association of MITE-like elements with Ae. aegypti genes shows the same bias toward noncoding regions as in plants. This association has potentially important implications for the evolution of gene regulation. PMID- 9207117 TI - hCTR1: a human gene for copper uptake identified by complementation in yeast. AB - The molecular mechanisms responsible for the cellular uptake of copper in mammalian cells are unknown. We describe isolation of a human gene involved in this process by complementation of the yeast high-affinity copper uptake mutant, ctr1. Besides complementing ctr1 growth defect on nonfermentable media, the human gene also rescues iron transport and SOD1 defects in ctr1 yeast. Overexpression of the gene in yeast leads to vulnerability to the toxicity of copper overload. In addition, its expression in ctr1 yeast significantly increases the level of cellular copper, as demonstrated by atomic absorption. We propose this gene as a candidate for high-affinity copper uptake in humans and by analogy have named it hCTR1. The hCTR1 and yeast CTR1 predicted transmembrane proteins are 29% identical, but the human protein is substantially smaller in both the extracellular metal-binding and intracellular domains. An additional human gene similar to hCTR1, here named hCTR2, was identified in a database search. Both hCTR1 and hCTR2 are expressed in all human tissues examined, and both genes are located in 9q31/32. These studies, together with the previously recognized functional and sequence similarity between the Menkes/Wilson copper export proteins and CCC2 in yeast, demonstrate that similar copper homeostatic mechanisms are used in these evolutionarily divergent organisms. PMID- 9207118 TI - Identification and characterization of Saccharomyces cerevisiae EXO1, a gene encoding an exonuclease that interacts with MSH2. AB - A two-hybrid screen was used to identify Saccharomyces cerevisiae genes encoding proteins that interact with MSH2. One gene was found to encode a homologue of Schizosaccharomyces pombe EXO1, a double-stranded DNA-specific 5'-3' exonuclease. S. cerevisiae EXO1 interacted with both S. cerevisiae and human MSH2 in two hybrid and coimmunoprecipitation experiments. exo1 mutants showed a mutator phenotype, and epistasis analysis was consistent with EXO1 functioning in the MSH2-dependent mismatch repair pathway. exo1 mutations were lethal in combination with rad27 mutations, and overexpression of EXO1 suppressed both the temperature sensitive and mutator phenotypes of rad27 mutants. PMID- 9207119 TI - Cloning of a novel T-cell protein FYB that binds FYN and SH2-domain-containing leukocyte protein 76 and modulates interleukin 2 production. AB - T cell receptor zeta (TcRzeta)/CD3 ligation initiates a signaling cascade that involves src kinases p56(lck) and zeta-associated protein 70, leading to the phosphorylation of substrates such as TcRzeta, Vav, SH2-domain-containing leukocyte protein 76 (SLP-76), cbl, and p120/130. FYN binding protein (FYB or p120/130) associates with p59(fyn), the TcRzeta/CD3 complex, and becomes tyrosine phosphorylated in response to receptor ligation. In this study, we report the cDNA cloning of human and murine FYB and show that it is restricted in expression to T cells and myeloid cells and possesses an overall unique hydrophilic sequence with several tyrosine-based motifs, proline-based type I and type II SH3 domain binding motifs, several putative lysine/glutamic acid-rich nuclear localization motifs, and a SH3-like domain. In addition to binding the src kinase p59(fyn), FYB binds specifically to the hematopoietic signaling protein SLP-76, an interaction mediated by the SLP-76 SH2 domain. In keeping with this, expression of FYB augmented interleukin 2 secretion from a T cell hybridoma, DC27.10, in response to TcRzeta/CD3 ligation. FYB is therefore a novel hematopoietic protein that acts as a component of the FYN and SLP-76 signaling cascades in T cells. PMID- 9207120 TI - Efficient loading of HLA-DR with a T helper epitope by genetic exchange of CLIP. AB - The HLA class II-associated invariant chain (Ii)-derived peptide (CLIP) occupies the peptide binding groove during assembly in the endoplasmic reticulum, travels with HLA class II to endosomal compartments, and is subsequently released to allow binding of antigenic peptides. We investigated whether the exchange of CLIP with a known T helper epitope at the DNA level would lead to efficient loading of this helper epitope onto HLA class II. For this purpose, a versatile Ii-encoding expression vector was created in which CLIP can be replaced with a helper epitope of choice. Upon supertransfection of HLA-DR1-transfected 293 cells with an Ii vector encoding a known T helper epitope (HA307-319), predominantly length variants of this epitope were detected in association with the HLA-DR1 molecules of these cells. Moreover, this transfectant was efficiently recognized by a peptide-specific T helper clone (HA1.7). The results suggest that this type of Ii vector can be used to create potent class II+ cellular vaccines in which defined T cell epitopes are continuously synthesized. PMID- 9207121 TI - Recombinant parvovirus-like particles as an antigen carrier: a novel nonreplicative exogenous antigen to elicit protective antiviral cytotoxic T cells. AB - To develop a strategy that promotes efficient antiviral immunity, hybrid virus like particles (VLP) were prepared by self-assembly of the modified porcine parvovirus VP2 capsid protein carrying a CD8(+) T cell epitope from the lymphocytic choriomeningitis virus nucleoprotein. Immunization of mice with these hybrid pseudoparticles, without adjuvant, induced strong cytotoxic T lymphocyte (CTL) responses against both peptide-coated- or virus-infected-target cells. This CD8(+) class I-restricted cytotoxic activity persisted in vivo for at least 9 months. Furthermore, the hybrid parvovirus-like particles were able to induce a complete protection of mice against a lethal lymphocytic choriomeningitis virus infection. To our knowledge, this study represents the first demonstration that hybrid nonreplicative VLP carrying a single viral CTL epitope can induce protection against a viral lethal challenge, in the absence of any adjuvant. These recombinant particles containing a single type of protein are easily produced by the baculovirus expression system and, therefore, represent a promising and safe strategy to induce strong CTL responses for the elimination of virus-infected cells. PMID- 9207122 TI - Homodimerization of tumor-reactive monoclonal antibodies markedly increases their ability to induce growth arrest or apoptosis of tumor cells. AB - Monoclonal antibodies (mAbs) that exert antitumor activity can do so by virtue of their effector function and/or their ability to signal growth arrest or cell death. In this study, we demonstrate that mAbs which have little or no signaling activity-i.e., anti-CD19, CD20, CD21, CD22 and Her-2-can become potent antitumor agents when they are converted into IgG-IgG homodimers. The homodimers exert antigrowth activity by signaling G0/G1 arrest or apoptosis, depending upon which cell surface molecule they bind. This activity is specific and, in the case of the anti-CD19 mAb, did not require an Fc portion. These results offer the possibility that homodimers of other tumor-reactive mAbs which have little antitumor activity as monomers might be potent, antitumor agents. PMID- 9207123 TI - Proteasome regulation of activation-induced T cell death. AB - Lactacystin, a microbial metabolite that inhibits protease activity only in the proteasome, was used to study the role of the proteasome in the activation induced cell death (AICD) of T cells. Lactacystin induces DNA fragmentation and apoptosis in a T cell hybridoma (DO.11. 10) in a dose-dependent manner. Between 1 and 10 microM, the mildly cytotoxic lactacystin inhibited the AICD of DO.11.10 cells cultured in anti-CD3-coated wells. Degradation of IkappaBbeta and the translocation of the NF-kappaB (p50/RelA) into the nucleus, which occurred at 1.5 hr after anti-CD3 activation, were inhibited by lactacystin. Lactacystin did not inhibit the expression of nuclear transcription factor Oct-1. The activation induced expression of the immediate-early gene, Nur77, and the T cell death genes, CD95 (Fas) and CD95 ligand (FasL), were inhibited. Functional expression of FasL cytotoxicity and the increase of cell surface Fas were also inhibited. Lactacystin must be added within 2 hr of activation to efficiently block AICD. In addition, lactacystin failed to inhibit the killing of DO.11.10 by FasL expressing allo-specific cytotoxic effector cells. These observations strongly suggest a direct link between the proteasome-dependent degradation of IkappaBbeta and the AICD that occurs through activation of the FasL gene and up-regulation of the Fas gene. PMID- 9207124 TI - The solution structure of the N-terminal domain of alpha2-macroglobulin receptor associated protein. AB - The three-dimensional structure of the N-terminal domain (residues 18-112) of alpha2-macroglobulin receptor-associated protein (RAP) has been determined by NMR spectroscopy. The structure consists of three helices composed of residues 23-34, 39-65, and 73-88. The three helices are arranged in an up-down-up antiparallel topology. The C-terminal 20 residues were shown not to be in a well defined conformation. A structural model for the binding of RAP to the family of low density lipoprotein receptors is proposed. It defines a role in binding for both the unordered C terminus and the structural scaffold of the core structure. Pathogenic epitopes for the rat disease Heymann nephritis, an experimental model of human membranous glomerulonephritis, have been identified in RAP and in the large endocytic receptor gp330/megalin. Here we provide the three-dimensional structure of the pathogenic epitope in RAP. The amino acid residues known to form the epitope are in a helix-loop-helix conformation, and from the structure it is possible to rationalize the published results obtained from studies of fragments of the N-terminal domain. PMID- 9207125 TI - A new class of obesity genes encodes leukocyte adhesion receptors. AB - Obesity is a complex disease, and multiple genes contribute to the trait. The description of five genes (ob, db, tub, Ay, and fat) responsible for distinct syndromes of spontaneous monogenic obesity in mice has advanced our knowledge of the genetics of obesity. However, many other genes involved in the expression of this disease remain to be determined. We report here the identification of an additional class of genes involved in the regulation of adipose tissue mass. These genes encode receptors mediating leukocyte adhesion. Mice deficient in intercellular adhesion molecule-1 became spontaneously obese in old age on normal mouse chow or at a young age when provided with a diet rich in fat. Mice deficient in the counterreceptor for intercellular adhesion molecule-1, the leukocyte integrin alphaMbeta2 (Mac-1), showed a similar obesity phenotype. Since all mice consumed approximately the same amount of food as controls, the leukocyte function appears to be in regulating lipid metabolism and/or energy expenditure. Our results indicate that (i) leukocytes play a role in preventing excess body fat deposition and (ii) defects in leukocyte adhesion receptors can result in obesity. PMID- 9207126 TI - Nuclear translocation of NF-kappaB is increased in dopaminergic neurons of patients with parkinson disease. AB - Evidence from postmortem studies suggest an involvement of oxidative stress in the degeneration of dopaminergic neurons in Parkinson disease (PD) that have recently been shown to die by apoptosis, but the relationship between oxidative stress and apoptosis has not yet been elucidated. Activation of the transcription factor NF-kappaB is associated with oxidative stress-induced apoptosis in several nonneuronal in vitro models. To investigate whether it may play a role in PD, we looked for the translocation of NF-kappaB from the cytoplasm to the nucleus, evidence of its activation, in melanized neurons in the mesencephalon of postmortem human brain from five patients with idiopathic PD and seven matched control subjects. In PD patients, the proportion of dopaminergic neurons with immunoreactive NF-kappaB in their nuclei was more than 70-fold that in control subjects. A possible relationship between the nuclear localization of NF-kappaB in mesencephalic neurons of PD patients and oxidative stress in such neurons has been shown in vitro with primary cultures of rat mesencephalon, where translocation of NF-kappaB is preceded by a transient production of free radicals during apoptosis induced by activation of the sphingomyelin-dependent signaling pathway with C2-ceramide. The data suggest that this oxidant-mediated apoptogenic transduction pathway may play a role in the mechanism of neuronal death in PD. PMID- 9207127 TI - Potentially predictive and manipulable blood serum correlates of aging in the healthy human male: progressive decreases in bioavailable testosterone, dehydroepiandrosterone sulfate, and the ratio of insulin-like growth factor 1 to growth hormone. AB - A cross-sectional survey was made in 56 exceptionally healthy males, ranging in age from 20 to 84 years. Measurements were made of selected steroidal components and peptidic hormones in blood serum, and cognitive and physical tests were performed. Of those blood serum variables that gave highly significant negative correlations with age (r > -0.6), bioavailable testosterone (BT), dehydroepiandrosterone sulfate (DHEAS), and the ratio of insulin-like growth factor 1 (IGF-1) to growth hormone (GH) showed a stepwise pattern of age-related changes most closely resembling those of the age steps themselves. Of these, BT correlated best with significantly age-correlated cognitive and physical measures. Because DHEAS correlated well with BT and considerably less well than BT with the cognitive and physical measures, it seems likely that BT and/or substances to which BT gives rise in tissues play a more direct role in whatever processes are rate-limiting in the functions measured and that DHEAS relates more indirectly to these functions. The high correlation of IGF-1/GH with age, its relatively low correlation with BT, and the patterns of correlations of IGF-1/GH and BT with significantly age-correlated cognitive and physical measures suggest that the GH-IGF-1 axis and BT play independent roles in affecting these functions. Serial determinations made after oral ingestion of pregnenolone and data from the literature suggest there is interdependence of steroid metabolic systems with those operational in control of interrelations in the GH-IGF-1 axis. Longitudinal concurrent measurements of serum levels of BT, DHEAS, and IGF-1/GH together with detailed studies of their correlations with age-correlated functional measures may be useful in detecting early age-related dysregulations and may be helpful in devising ameliorative approaches. PMID- 9207128 TI - Angiotensin II type1a receptor gene expression in the heart: AP-1 and GATA-4 participate in the response to pressure overload. AB - Hypertrophy of mammalian cardiac muscle is mediated, in part, by angiotensin II through an angiotensin II type1a receptor (AT1aR)-dependent mechanism. To understand how the level of AT1aRs is altered in this pathological state, we studied the expression of an injected AT1aR promoter-luciferase reporter gene in adult rat hearts subjected to an acute pressure overload by aortic coarctation. This model was validated by demonstrating that coarctation increased expression of the alpha-skeletal actin promoter 1.7-fold whereas the alpha-myosin heavy chain promoter was unaffected. Pressure overload increased expression from the AT1aR promoter by 1. 6-fold compared with controls. Mutations introduced into consensus binding sites for AP-1 or GATA transcription factors abolished the pressure overload response but had no effect on AT1aR promoter activity in control animals. In extracts from coarcted hearts, but not from control hearts, a Fos-JunB-JunD complex and GATA-4 were detected in association with the AP-1 and GATA sites, respectively. These results establish that the AT1aR promoter is active in cardiac muscle and its expression is induced by pressure overload, and suggest that this response is mediated, in part, by a functional interaction between AP-1 and GATA-4 transcription factors. PMID- 9207130 TI - Reduced fertility in mice deficient for the POU protein sperm-1. AB - Members of the POU-homeodomain gene family encode transcriptional regulatory molecules that play important roles in terminal differentiation of many organ systems. Sperm-1 (Sprm-1) is a POU domain factor that is exclusively expressed in the differentiating male germ cell. We show here that the Sprm-1 protein is expressed in the haploid spermatid and that 129/Sv Sprm-1(-/-) mice are subfertile when compared with wild-type or heterozygous littermates yet exhibit normal testicular morphology and produce normal numbers of mobile spermatozoa. Our data suggest that the Sprm-1 protein plays a discrete regulatory function in the haploid spermatid, which is required for the optimal function, but not the terminal differentiation, of the male germ cell. PMID- 9207129 TI - Gestational exposure to ethanol suppresses msx2 expression in developing mouse embryos. AB - Ethanol acts as a teratogen in developing fetuses causing abnormalities of the brain, heart, craniofacial bones, and limb skeletal elements. To assess whether some teratogenic actions of ethanol might occur via dysregulation of msx2 expression, we examined msx2 expression in developing mouse embryos exposed to ethanol on embryonic day (E) 8 of gestation and subjected to whole mount in situ hybridization on E11-11.5 using a riboprobe for mouse msx2. Control mice exhibited expression of msx2 in developing brain, the developing limb buds and apical ectodermal ridge, the lateral and nasal processes, olfactory pit, palatal shelf of the maxilla, the eye, the lens of the eye, otic vesicle, prevertebral bodies (notochord), and endocardial cushion. Embryos exposed to ethanol in utero were significantly smaller than their normal counterparts and did not exhibit expression of msx2 in any structures. Similarly, msx2 expression, as determined by reverse transcription-PCR and Northern blot hybridization, was reduced approximately 40-50% in fetal mouse calvarial osteoblastic cells exposed to 1% ethanol for 48 hr while alkaline phosphatase was increased by 2-fold and bone morphogenetic protein showed essentially no change. Transcriptional activity of the msx2 promoter was specifically suppressed by alcohol in MC3T3-E1 osteoblasts. Taken together, these data demonstrate that fetal alcohol exposure decreases msx2 expression, a known regulator of osteoblast and myoblast differentiation, and suggest that one of the "putative" mechanisms for fetal alcohol syndrome is the inhibition of msx2 expression during key developmental periods leading to developmental retardation, altered craniofacial morphogenesis, and cardiac defects. PMID- 9207131 TI - Up-regulation of specific tyrosinase mRNAs in mouse melanomas with the c2j gene substituted for the wild-type tyrosinase allele: utilization in design of syngeneic immunotherapy models. AB - The expression of cell-specialization genes is likely to be changing in tumor cells as their differentiation declines. Functional changes in these genes might yield unusual peptide epitopes with anti-tumor potential and could occur without modification in the DNA sequence of the gene. Melanomas undergo a characteristic decline in melanization that may reflect altered contributions of key melanocytic genes such as tyrosinase. Quantitative reverse transcriptase-PCR of the wild-type (C) tyrosinase gene in transgenic (C57BL/6 strain) mouse melanomas has revealed a shift toward alternative splicing of the pre-mRNA that generated increased levels of the Delta1b and Delta1d mRNA splice variants. The spontaneous c2j albino mutation of tyrosinase (in the C57BL/6 strain) changes the pre-mRNA splicing pattern. In c2j/c2j melanomas, alternative splicing was again increased. However, while some mRNAs (notably Delta1b) present in C/C were obligatorily absent, others (Delta3 and Delta1d) were elevated. In c2j/c2j melanomas, the percentage of total tyrosinase transcripts attributable to Delta3 reached approximately 2 fold the incidence in c2j/c2j or C/C skin melanocytes. The percentage attributable to Delta1d rose to approximately 2-fold the incidence in c2j/c2j skin, and to 10-fold that in C/C skin. These differences provide a basis for unique mouse models in which the melanoma arises in skin grafted from a C/C or c2j/c2j transgenic donor to a transgenic host of the same or opposite tyrosinase genotype. Immunotherapy designs then could be based on augmenting those antigenic peptides that are novel or overrepresented in a tumor relative to the syngeneic host. PMID- 9207132 TI - Chemical camouflage of antigenic determinants: stealth erythrocytes. AB - In a number of clinical circumstances it would be desirable to artificially conceal cellular antigenic determinants to permit survival of heterologous donor cells. A case in point is the problem encountered in transfusions of patients with rare blood types or chronically transfused patients who become allosensitized to minor blood group determinants. We have tested the possibility that chemical modification of the red blood cell (RBC) membrane might serve to occlude antigenic determinants, thereby minimizing transfusion reactions. To this end, we have covalently bound methoxy(polyethylene glycol) (mPEG) to the surface of mammalian RBC via cyanuric chloride coupling. Human RBC treated with this technique lose ABO blood group reactivity as assessed by solution-phase antisera agglutination. In accord with this, we also find a profound decrease in anti blood group antibody binding. Furthermore, whereas human monocytes avidly phagocytose untreated sheep RBC, mPEG-derivatized sheep RBC are ineffectively phagocytosed. Surprisingly, human and mouse RBC appear unaffected by this covalent modification of the cell membrane. Thus, mPEG-treated RBC are morphologically normal, have normal osmotic fragility, and mPEG-derivatized murine RBC have normal in vivo survival, even following repeated infusions. Finally, in preliminary experiments, mPEG-modified sheep RBC intraperitoneally transfused into mice show significantly improved (up to 360-fold) survival when compared with untreated sheep RBC. We speculate that similar chemical camouflage of intact cells may have significant clinical applications in both transfusion (e.g., allosensitization and autoimmune hemolytic disease) and transplantation (e.g., endothelial cells and pancreatic beta cells) medicine. PMID- 9207133 TI - Transcription and activity of antioxidant enzymes after ionizing irradiation in radiation-resistant and radiation-sensitive mice. AB - The involvement of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase in radiobiological processes has been described at the enzyme activity level. We irradiated radiation-resistant (RR) and radiation-sensitive (RS) mice and studied antioxidant enzymes at the transcriptional and activity level. In addition, aromatic hydroxylation and lipid peroxidation parameters were determined to study radiation resistance at the oxidation level. RS BALB/c/J Him mice and RR C3H He/Him mice were whole-body irradiated with x-rays at 2, 4, and 6 Gy and killed 5, 15, and 30 min after irradiation. mRNA was isolated from liver and hybridized with probes for antioxidant enzymes and beta-actin as a housekeeping gene control. Antioxidant enzyme activities were determined by standard assays. Parameters for aromatic hydroxylation (o-tyrosine) and lipid peroxidation (malondialdehyde) were determined by HPLC methods. Antioxidant transcription was unchanged in contrast to antioxidant activities; SOD and CAT activities were elevated within 15 min in RR animals but not in RS mice, at all doses studied. Glutathione peroxidase activity was not different between RR and RS mice and was only moderately elevated after irradiation. No significant differences were found between RR and RS animals at the oxidation level, although a radiation dose-dependent increase of oxidation products was detected in both groups. We found that ionizing irradiation led to increased antioxidant activity only minutes after irradiation in the absence of increased transcription of these antioxidant enzymes. RR animals show higher antioxidant enzyme activities than do RS mice, but oxidation products are comparable in RS and RR mice. As unchanged transcription of antioxidant enzymes could not have been responsible for the increased antioxidant enzyme activities, preformed antioxidant enzymes should have been released by the irradiation process. This would be in agreement with previous studies of preformed, stored SOD. The finding of higher SOD and CAT activities in RR than in RS animals could point to a role for these antioxidant enzymes for the process of radiation sensitivity. PMID- 9207134 TI - Cloning, expression, and characterization of a cDNA encoding a novel human growth factor for primitive hematopoietic progenitor cells. AB - Multiple growth factors synergistically stimulate proliferation of primitive hematopoietic progenitor cells. A human myeloid cell line, KPB-M15, constitutively produces a novel hematopoietic cytokine, termed stem cell growth factor (SCGF), possessing species-specific proliferative activities. Here we report the molecular cloning, expression, and characterization of a cDNA encoding human SCGF using a newly developed lambdaSHDM vector that is more efficient for differential and expression cloning. cDNA for SCGF encodes a 29-kDa polypeptide without N-linked glycosylation. SCGF transiently produced by COS-1 cells supports growth of hematopoietic progenitor cells through a short-term liquid culture of bone marrow cells and exhibits promoting activities on erythroid and granulocyte/macrophage progenitor cells in primary semisolid culture with erythropoietin and granulocyte/macrophage colony-stimulating factor, respectively. Expression of SCGF mRNA is restricted to myeloid cells and fibroblasts, suggesting that SCGF is a growth factor functioning within the hematopoietic microenvironment. SCGF could disclose some human-specific mechanisms as yet unidentified from studies on the murine hematopoietic system. PMID- 9207135 TI - Molecular identification of a novel retrovirus repeatedly isolated from patients with multiple sclerosis. The Collaborative Research Group on Multiple Sclerosis. AB - The partial molecular characterization of multiple sclerosis (MS)-associated retrovirus (MSRV), a novel retrovirus previously called LM7, is reported. MSRV has been isolated repeatedly from leptomeningeal, choroid plexus and from Epstein Barr virus-immortalized B cells of MS patients. A strategy based on reverse transcriptase PCR with RNA-purified extracellular virions yielded an initial pol fragment from which other regions of the retroviral genome were subsequently obtained by sequence extension. MSRV-specific PCR primers amplified a pol region from RNA present at the peak of reverse transcriptase activity, coinciding with extracellular viral particles in sucrose density gradients. The same sequence was detected in noncellular RNA from MS patient plasma and in cerebrospinal fluid from untreated MS patients. MSRV is related to, but distinct from, the endogenous retroviral sequence ERV9. Whether MSRV represents an exogenous retrovirus with closely related endogenous elements or a replication-competent, virion-producing, endogenous provirus is as yet unknown. Further molecular epidemiological studies are required to determine precisely the apparent association of virions containing MSRV RNA with MS. PMID- 9207136 TI - GPIIIa-(49-66) is a major pathophysiologically relevant antigenic determinant for anti-platelet GPIIIa of HIV-1-related immunologic thrombocytopenia. AB - High-affinity (Kd = 1 x 10(-9) M) anti-platelet GPIIIa has been isolated from serum immune complexes of immunologic thrombocytopenic HIV-1-infected patients (HIV-1-ITP). Affinity-purified anti-platelet antibody reacted with a recombinant GPIIIa-(1-200) and -(1-66) fusion peptide and with an 18-mer GPIIIa-(49-66) peptide but not with seven other GPIIIa peptides spanning the length of GPIIIa. Most of the anti-platelet antibody ( approximately 85%) could be adsorbed to and eluted from a GPIIIa-(49-66) affinity column. Binding of antibody to platelets could be inhibited by GPIIIa-(49-66) or an equimolar peptide-albumin conjugate (IC50 = 2 microM). Sera from 7 control subjects and 10 classic autoimmune thrombocytopenic patients gave background reactivity with GPIIIa-(49-66). HIV-1 ITP sera from 16 patients reacted with a mean OD 6-fold greater than background (range, 4- to 9-fold). Serum anti-GPIIIa-(49-66) concentration correlated inversely with platelet count, R2 = 0.51, n = 31, P < 0. 0001. Because mouse platelet GPIIIa-(49-66) has 83% homology with human GPIIIa and mouse monocytes contain Fc receptors for the human IgG1-kappa/lambda antibody, we determined the in vivo effect of human anti-GPIIIa on mouse platelets. Affinity-purified antibody, 25-50 microg given i.p., resulted in a precipitous drop in platelet count to 30% of baseline, with nadir at 4 hr and return to normal in 36 hr. No effect was noted with control IgG. Acute thrombocytopenia could be prevented or reversed by the injection of the GPIIIa-(49-66) albumin conjugate at zero time or 2 hr after antibody, respectively, but not with a scrambled peptide-albumin conjugate. Thus HIV-1-ITP patients have high-affinity anti-platelet GPIIIa against a major antigenic determinant, GPIIIa-(49-66), which correlates inversely with platelet count and induces thrombocytopenia in mice. PMID- 9207137 TI - Induction of host signal transduction pathways by Helicobacter pylori. AB - Adherence of Helicobacter pylori to cultured gastric epithelial cells is associated with several cellular events, including the tyrosine phosphorylation of a 145-kDa host protein; the reorganization of the host cell actin and associated cellular proteins, like vasodilator-stimulated phosphoprotein, adjacent to the attached bacterial cell; and the subsequent release of the cytokine, interleukin 8 (IL-8). H. pylori isolated from patients with ulcer disease and gastric cancer contain a DNA insertion, the cag pathogenicity island (PAI), that is not present in bacteria isolated from individuals with asymptomatic infection. Mutations in a number of PAI genes abolish tyrosine phosphorylation and IL-8 synthesis but not the cytoskeletal rearrangements. Kinase inhibition studies suggest there are two distinct pathways operative in stimulating IL-8 release from host cells and one of these H. pylori pathways is independent of the tyrosine phosphorylation step. PMID- 9207138 TI - CD68+ cells of monocyte/macrophage lineage in the environment of AIDS-associated and classic-sporadic Kaposi sarcoma are singly or doubly infected with human herpesviruses 7 and 6B. AB - Earlier studies have shown that Kaposi sarcomas contain cells infected with human herpesvirus (HHV) 6B, and in current studies we report that both AIDS-associated and classic-sporadic Kaposi sarcoma contain HHV-7 genome sequences detectable by PCR. To determine the distribution of HHV-7-infected cells relative to those infected with HHV-6, sections from paraffin-embedded tissues were allowed to react with antibodies to HHV-7 virion tegument phosphoprotein pp85 and to HHV-6B protein p101. The antibodies are specific for HHV-7 and HHV-6B, respectively, and they retained reactivity for antigens contained in formalin-fixed, paraffin embedded tissue samples. We report that (i) HHV-7 pp85 was present in 9 of 32 AIDS-associated Kaposi sarcomas, and in 1 of 7 classical-sporadic HIV-negative Kaposi sarcomas; (ii) HHV-7 pp85 was detected primarily in cells bearing the CD68 marker characteristic of the monocyte/macrophage lineage present in or surrounding the Kaposi sarcoma lesions; and (iii) in a number of Kaposi sarcoma specimens, tumor-associated CD68+ monocytes/macrophages expressed simultaneously antigens from both HHV-7 and HHV-6B, and therefore appeared to be doubly infected with the two viruses. CD68+ monocytes/macrophages infected with HHV-7 were readily detectable in Kaposi sarcoma, but virtually absent from other normal or pathological tissues that harbor macrophages. Because all of the available data indicate that HHV-7 infects CD4+ T lymphocytes, these results suggest that the environment of the Kaposi sarcoma (i) attracts circulating peripheral lymphocytes and monocytes, triggers the replication of latent viruses, and thereby increases the local concentration of viruses, (ii) renders CD68+ monocytes/macrophages susceptible to infection with HHV-7, and (iii) the combination of both events enables double infections of cells with both HHV-6B and HHV-7. PMID- 9207140 TI - Aggregation and disaggregation of senile plaques in Alzheimer disease. AB - We quantitatively analyzed, using laser scanning confocal microscopy, the three dimensional structure of individual senile plaques in Alzheimer disease. We carried out the quantitative analysis using statistical methods to gain insights about the processes that govern Abeta peptide deposition. Our results show that plaques are complex porous structures with characteristic pore sizes. We interpret plaque morphology in the context of a new dynamical model based on competing aggregation and disaggregation processes in kinetic steady-state equilibrium with an additional diffusion process allowing Abeta deposits to diffuse over the surface of plaques. PMID- 9207139 TI - Elevated free nitrotyrosine levels, but not protein-bound nitrotyrosine or hydroxyl radicals, throughout amyotrophic lateral sclerosis (ALS)-like disease implicate tyrosine nitration as an aberrant in vivo property of one familial ALS linked superoxide dismutase 1 mutant. AB - Mutations in superoxide dismutase 1 (SOD1; EC 1.15.1.1) are responsible for a proportion of familial amyotrophic lateral sclerosis (ALS) through acquisition of an as-yet-unidentified toxic property or properties. Two proposed possibilities are that toxicity may arise from imperfectly folded mutant SOD1 catalyzing the nitration of tyrosines [Beckman, J. S., Carson, M., Smith, C. D. & Koppenol, W. H. (1993) Nature (London) 364, 584] through use of peroxynitrite or from peroxidation arising from elevated production of hydroxyl radicals through use of hydrogen peroxide as a substrate [Wiedau-Pazos, M., Goto, J. J., Rabizadeh, S., Gralla, E. D., Roe, J. A., Valentine, J. S. & Bredesen, D. E. (1996) Science 271, 515-518]. To test these possibilities, levels of nitrotyrosine and markers for hydroxyl radical formation were measured in two lines of transgenic mice that develop progressive motor neuron disease from expressing human familial ALS linked SOD1 mutation G37R. Relative to normal mice or mice expressing high levels of wild-type human SOD1, 3-nitrotyrosine levels were elevated by 2- to 3-fold in spinal cords coincident with the earliest pathological abnormalities and remained elevated in spinal cord throughout progression of disease. However, no increases in protein-bound nitrotyrosine were found during any stage of SOD1-mutant mediated disease in mice or at end stage of sporadic or SOD1-mediated familial human ALS. When salicylate trapping of hydroxyl radicals and measurement of levels of malondialdehyde were used, there was no evidence throughout disease progression in mice for enhanced production of hydroxyl radicals or lipid peroxidation, respectively. The presence of elevated nitrotyrosine levels beginning at the earliest stages of cellular pathology and continuing throughout progression of disease demonstrates that tyrosine nitration is one in vivo aberrant property of this ALS-linked SOD1 mutant. PMID- 9207141 TI - Functional impact of cerebral connections. AB - Cerebral networks are complex sets of connections that resemble a ladder-like web of multiple parallel feedforward, lateral, and feedback connections. This static anatomical description has been pivotal in guiding our understanding of signal processing within cerebral networks. However, measures on both magnitude and functional significance of connections are extremely limited. Here, we compare the anatomically defined strengths of a set of cerebral pathways emerging from the visual middle suprasylvian (MS) cortex of the cat with measures of the functional impact the same region has over distant sites. These functional measures were obtained by analyzing the local and distant effects of MS cooling deactivation on deoxyglucose uptake. Relative to major efferent projections from MS cortex that have a strong influence, projections to early visual processing stages have weaker functional influences than predicted from the anatomy. For higher processing stages, the converse holds: projections from MS cortex have stronger functional influence than predicted from the anatomy. We conclude that these and future functional measures, obtained using the same combination of techniques, will furnish fundamental, new information that complements and extends current models of static cerebral networks, and lead to more realistic models of cerebral network function and component interactions. PMID- 9207142 TI - Visual stimuli induce waves of electrical activity in turtle cortex. AB - The computations involved in the processing of a visual scene invariably involve the interactions among neurons throughout all of visual cortex. One hypothesis is that the timing of neuronal activity, as well as the amplitude of activity, provides a means to encode features of objects. The experimental data from studies on cat [Gray, C. M., Konig, P., Engel, A. K. & Singer, W. (1989) Nature (London) 338, 334-337] support a view in which only synchronous (no phase lags) activity carries information about the visual scene. In contrast, theoretical studies suggest, on the one hand, the utility of multiple phases within a population of neurons as a means to encode independent visual features and, on the other hand, the likely existence of timing differences solely on the basis of network dynamics. Here we use widefield imaging in conjunction with voltage sensitive dyes to record electrical activity from the virtually intact, unanesthetized turtle brain. Our data consist of single-trial measurements. We analyze our data in the frequency domain to isolate coherent events that lie in different frequency bands. Low frequency oscillations (<5 Hz) are seen in both ongoing activity and activity induced by visual stimuli. These oscillations propagate parallel to the afferent input. Higher frequency activity, with spectral peaks near 10 and 20 Hz, is seen solely in response to stimulation. This activity consists of plane waves and spiral-like waves, as well as more complex patterns. The plane waves have an average phase gradient of approximately pi/2 radians/mm and propagate orthogonally to the low frequency waves. Our results show that large-scale differences in neuronal timing are present and persistent during visual processing. PMID- 9207143 TI - Positron-emission tomography imaging of long-term shape recognition challenges. AB - Long-term visual memory performance was impaired by two types of challenges: a diazepam challenge on acquisition and a sensory challenge on recognition. Using positron-emission tomography regional cerebral blood flow imaging, we studied the effect of these challenges on regional brain activation during the delayed recognition of abstract visual shapes as compared with a baseline fixation task. Both challenges induced a significant decrease in differential activation in the left fusiform gyrus, suggesting that this region is involved in the automatic or volitional comparison of incoming and stored stimuli. In contrast, thalamic differential activation increased in response to memory challenges. This increase might reflect enhanced retrieval attempts as a compensatory mechanism for restoring recognition performance. PMID- 9207144 TI - Transmembrane topology of a CLC chloride channel. AB - CLC chloride channels form a large and conserved gene family unrelated to other channel proteins. Knowledge of the transmembrane topology of these channels is important for understanding the effects of mutations found in human myotonia and inherited hypercalciuric kidney stone diseases and for the interpretation of structure-function studies. We now systematically study the topology of human ClC 1, a prototype CLC channel that is defective in human myotonia. Using a combination of in vitro glycosylation scanning and protease protection assays, we show that both N and C termini face the cytoplasm and demonstrate the presence of 10 (or less likely 12) transmembrane spans. Difficult regions were additionally tested by inserting cysteines and probing the effect of cysteine-modifying reagents on ClC-1 currents. The results show that D3 crosses the membrane and D4 does not, and that L549 between D11 and D12 is accessible from the outside. Further, since the modification of cysteines introduced between D11 and D12 and at the extracellular end of D3 strongly affect ClC-1 currents, these regions are suggested to be important for ion permeation. PMID- 9207145 TI - A single serine residue controls the cation dependence of substrate transport by the rat serotonin transporter. AB - The serotonin transporter (SERT) is a member of the Na+/Cl--dependent neurotransmitter transporter family and constitutes the target of several clinically important antidepressants. Here, replacement of serine-545 in the recombinant rat SERT by alanine was found to alter the cation dependence of serotonin uptake. Substrate transport was now driven as efficiently by LiCl as by NaCl without significant changes in serotonin affinity. Binding of the antidepressant [3H]imipramine occurred with 1/5th the affinity, whereas [3H]citalopram binding was unchanged. These results indicate that serine-545 is a crucial determinant of both the cation dependence of serotonin transport by SERT and the imipramine binding properties of SERT. PMID- 9207146 TI - Head direction cells and episodic spatial information in rats without a hippocampus. AB - To successfully navigate through the environment animals rely on information concerning their directional heading and location. Many cells within the postsubiculum and anterior thalamus discharge as a function of the animal's head direction (HD), while many cells in the hippocampus discharge in relation to the animal's location. We placed lesions in the hippocampus and recorded from HD cells in the postsubiculum and anterior thalamus. Lesions of the hippocampus did not disrupt the HD cell signal in either brain area, indicating that the HD cell signal must be generated by structures external to the hippocampus. In addition, each cell's preferred firing direction remained stable across days when the lesioned animal was placed into a novel environment. This stability appeared to weaken after several weeks of nonexposure to the new enclosure for two out of five animals, and subsequently recorded cells from these two animals established a new angular relationship between the familiar and novel environments. Our results suggest that extra-hippocampal structures are capable of creating and maintaining a novel representation of the animal's environmental context. This representation shares features in common with mnemonic processes involving episodic memory that until now were assumed to require an intact hippocampus. PMID- 9207147 TI - Streaming potential measurements in alphabetagamma-rat epithelial Na+ channel in planar lipid bilayers. AB - Streaming potentials across cloned epithelial Na+ channels (ENaC) incorporated into planar lipid bilayers were measured. We found that the establishment of an osmotic pressure gradient (Deltapi) across a channel-containing membrane mimicked the activation effects of a hydrostatic pressure differential (DeltaP) on alphabetagamma-rENaC, although with a quantitative difference in the magnitude of the driving forces. Moreover, the imposition of a Deltapi negates channel activation by DeltaP when the Deltapi was directed against DeltaP. A streaming potential of 2.0 +/- 0.7 mV was measured across alphabetagamma-rat ENaC (rENaC) containing bilayers at 100 mM symmetrical [Na+] in the presence of a 2 Osmol/kg sucrose gradient. Assuming single file movement of ions and water within the conduction pathway, we conclude that between two and three water molecules are translocated together with a single Na+ ion. A minimal effective pore diameter of 3 A that could accommodate two water molecules even in single file is in contrast with the 2-A diameter predicted from the selectivity properties of alphabetagamma rENaC. The fact that activation of alphabetagamma-rENaC by DeltaP can be reproduced by the imposition of Deltapi suggests that water movement through the channel is also an important determinant of channel activity. PMID- 9207148 TI - Slow cycling of unphosphorylated myosin is inhibited by calponin, thus keeping smooth muscle relaxed. AB - A key unanswered question in smooth muscle biology is whether phosphorylation of the myosin regulatory light chain (RLC) is sufficient for regulation of contraction, or if thin-filament-based regulatory systems also contribute to this process. To address this issue, the endogenous RLC was extracted from single smooth muscle cells and replaced with either a thiophosphorylated RLC or a mutant RLC (T18A/S19A) that cannot be phosphorylated by myosin light chain kinase. The actin-binding protein calponin was also extracted. Following photolysis of caged ATP, cells without calponin that contained a nonphosphorylatable RLC shortened at 30% of the velocity and produced 65% of the isometric force of cells reconstituted with the thiophosphorylated RLC. The contraction of cells reconstituted with nonphosphorylatable RLC was, however, specifically suppressed in cells that contained calponin. These results indicate that calponin is required to maintain cells in a relaxed state, and that in the absence of this inhibition, dephosphorylated cross-bridges can slowly cycle and generate force. These findings thus provide a possible framework for understanding the development of latch contraction, a widely studied but poorly understood feature of smooth muscle. PMID- 9207149 TI - Ceramide-induced inhibition of T lymphocyte voltage-gated potassium channel is mediated by tyrosine kinases. AB - The n-type K+ channel (n-K+, Kv1.3) in lymphocytes has been recently implicated in the regulation of Fas-induced programmed cell death. Here, we demonstrate that ceramide, a lipid metabolite synthesized upon Fas receptor ligation, inhibits n K+ channel activity and induces a tyrosine phosphorylation of the Kv1.3 protein in Jurkat T lymphocytes. Tyrosine phosphorylation of the n-K+ channel correlated with an activation of the Src-like tyrosine kinase p56lck upon cellular treatment with the ceramide analog C6-ceramide. Because genetic deficiency of p56lck or inhibition of Src-like tyrosine kinases by herbimycin A prevented ceramide mediated n-K+ channel inhibition and tyrosine phosphorylation, we propose a ceramide-initiated activation of p56lck resulting in tyrosine phosphorylation and inhibition of the n-K+ channel protein. PMID- 9207151 TI - Striking sequence similarity in inter- and intra-specific comparisons of class I SLG alleles from Brassica oleracea and Brassica campestris: implications for the evolution and recognition mechanism. AB - Self-incompatibility in Brassica is controlled by a single multi-allelic locus (S locus), which contains at least two highly polymorphic genes expressed in the stigma: an S glycoprotein gene (SLG) and an S receptor kinase gene (SRK). The putative ligand-binding domain of SRK exhibits high homology to the secretory protein SLG, and it is believed that SLG and SRK form an active receptor kinase complex with a self-pollen ligand, which leads to the rejection of self-pollen. Here, we report 31 novel SLG sequences of Brassica oleracea and Brassica campestris. Sequence comparisons of a large number of SLG alleles and SLG-related genes revealed the following points. (i) The striking sequence similarity observed in an inter-specific comparison (95.6% identity between SLG14 of B. oleracea and SLG25 of B. campestris in deduced amino acid sequence) suggests that SLG diversification predates speciation. (ii) A perfect match of the sequences in hypervariable regions, which are thought to determine S specificity in an intra specific comparison (SLG8 and SLG46 of B. campestris) and the observation that the hypervariable regions of SLG and SRK of the same S haplotype were not necessarily highly similar suggests that SLG and SRK bind different sites of the pollen ligand and that they together determine S specificity. (iii) Comparison of the hypervariable regions of SLG alleles suggests that intragenic recombination, together with point mutations, has contributed to the generation of the high level of sequence variation in SLG alleles. Models for the evolution of SLG/SRK are presented. PMID- 9207153 TI - A maize zinc-finger protein binds the prolamin box in zein gene promoters and interacts with the basic leucine zipper transcriptional activator Opaque2. AB - The prolamin box (P-box) is a highly conserved 7-bp sequence element (5'-TGTAAAG 3') found in the promoters of many cereal seed storage protein genes. Nuclear factors from maize endosperm specifically interact with the P-box present in maize prolamin genes (zeins). The presence of the P-box in all zein gene promoters suggests that interactions between endosperm DNA binding proteins and the P-box may play an important role in the coordinate activation of zein gene expression during endosperm development. We have cloned an endosperm-specific maize cDNA, named prolamin-box binding factor (PBF), that encodes a member of the recently described Dof class of plant Cys2-Cys2 zinc-finger DNA binding proteins. When tested in gel shift assays, PBF exhibits the same sequence-specific binding to the P-box as factors present in maize endosperm nuclei. Additionally, PBF interacts in vitro with the basic leucine zipper protein Opaque2, a known transcriptional activator of zein gene expression whose target site lies 20 bp downstream of the P-box in the 22-kDa zein gene promoter. The isolation of the PBF gene provides an essential tool to further investigate the functional role of the highly conserved P-box in regulating cereal storage protein gene expression. PMID- 9207152 TI - A reversibly glycosylated polypeptide (RGP1) possibly involved in plant cell wall synthesis: purification, gene cloning, and trans-Golgi localization. AB - We purified from pea (Pisum sativum) tissue an approximately 40 kDa reversibly glycosylated polypeptide (RGP1) that can be glycosylated by UDP-Glc, UDP-Xyl, or UDP-Gal, and isolated a cDNA encoding it, apparently derived from a single-copy gene (Rgp1). Its predicted translation product has 364 aminoacyl residues and molecular mass of 41.5 kDa. RGP1 appears to be a membrane-peripheral protein. Immunogold labeling localizes it specifically to trans-Golgi dictyosomal cisternae. Along with other evidence, this suggests that RGP1 is involved in synthesis of xyloglucan and possibly other hemicelluloses. Corn (Zea mays) contains a biochemically similar and structurally homologous RGP1, which has been thought (it now seems mistakenly) to function in starch synthesis. The expressed sequence database also reveals close homologs of pea Rgp1 in Arabidopsis and rice (Oryza sativa). Rice possesses, in addition, a distinct but homologous sequence (Rgp2). RGP1 provides a polypeptide marker for Golgi membranes that should be useful in plant membrane studies. PMID- 9207154 TI - Decompression is essential in the management of small bowel obstruction. PMID- 9207155 TI - Effect of balloon-expandable and self-expanding stent fixation on endoluminal polytetrafluoroethylene graft healing. AB - PURPOSE: To investigate the effect of stent design and deployment mechanism on endoluminal graft healing. METHOD: Twenty dogs underwent infrarenal abdominal aorta polytetrafluoroethylene (PTFE) interposition (6) or intraluminal stented grafting using either a balloon expandable (BE, n = 8) or self-expanding (SE, n = 6) stent design. Grafts were removed at 8 weeks. Length of endothelial ingrowth and intima to media height ratios (IMHR) were calculated. Perianastomotic smooth muscle (Actin+), macrophage (CD44+), proliferating (PCNA+), and platelet-derived growth factor (PDGF+) cell content were determined. RESULTS: Mean endothelial ingrowth was 1.10 +/- 0.15 cm (control), 1.88 +/- 0.13 cm (BESG), and 2.16 +/- 0.18 cm (SESG) proximally; and 0.94 +/- 0.12, 2.11 +/- 0.11 cm, and 2.16 +/- 0.15 cm, respectively, at the distal anastomosis. Endothelial ingrowth was greater in all stented grafts (P <0.001). Mean IMHRs were 1.42 +/- 0.16 (control), 0.50 +/- 0.14 (BESG), and 0.77 +/- 0.2 (SESG) proximally; and 0.84 +/- 0.1, 0.42 +/- 0.09, and 0.77 +/- 0.12 (SESG) distally. Lower IMHRs were observed in all stented graft regions (P <0.05) except the distal anastomosis of SESG. The PDGF+ and PCNA+ cell content was decreased, and Actin+ cell content was increased in all stented grafts (P <0.05). CONCLUSION: Intraluminal location enhances endothelialization and attenuates intimal thickening in PTFE grafts. The enhanced healing of intraluminal stented grafts is irrespective of the type of stent or deployment mechanism used. PMID- 9207156 TI - A study of prognostic factors for hepatic resection for colorectal metastases. AB - BACKGROUND: Liver resection is accepted treatment for selected patients with colon cancer metastatic to the liver. There remains some controversy regarding the selection criteria, particularly which preoperative features are useful predictors of long survival postresection. METHODS: One hundred and twenty-three patients who had liver resection for colorectal metastases on the Hepato Pancreatic Biliary Service at The Toronto Hospital between August 1977 and June 1993 were studied. Seventy-seven had solitary lesions, 15 had single lesions with satellite nodules, and 31 had multiple lesions. Synchronous liver metastases were found in 40 patients and 83 patients had metachronous lesions. Fifty-one patients had formal lobectomies and 21 had extended lobectomies. RESULTS: Postoperative complications were seen in 28% of patients, but there were no operative or postoperative deaths. Overall actuarial 5-year survival was 34%. There was a significant difference in survival according to the number of metastases. Patients with single lesions had a 5-year survival of 47% compared with 16% for single lesions with satellite nodules, and 17% for multiple lesions. There were no significant differences in survival based on age, sex, synchronous versus metachronous lesions, status of lymph nodes at the time of original surgery, intraoperative blood replacement, or size of tumor. CONCLUSIONS: An aggressive approach to the surgical management of colorectal liver metastases is possible with low risk in centers specializing in liver surgery, and results in prolonged survival in one third of patients. The most reliable predictor of long-term survival is the number of metastases in the liver. PMID- 9207157 TI - Postoperative analgesia reduces mortality and morbidity after esophagectomy. AB - BACKGROUND: To study the influence of postoperative analgesia on morbidity and mortality after esophagectomy. METHODS: The outcomes of 578 patients who underwent one-stage resection between 1986 and 1995 were analyzed. Patients who received either epidural morphine, patient-controlled analgesia, or continuous intravenous morphine infusion supervised by an anesthesiology-based acute pain service (group APS, n = 299) were compared with those for whom conventional intramuscular meperidine injections were used (group CON, n = 279). RESULTS: For patients who underwent transthoracic esophagectomy, group APS (n = 226) had a lower incidence of pulmonary complications (13% versus 25%, P = 0.002), cardiovascular complications (21% versus 43%, P < 0.001), and hospital mortality (8% versus 14%, P = 0.038) when compared with group CON (n = 189). No similar difference was demonstrated in patients who underwent esophagectomy without thoracotomy. The hospital stay (days) was shorter in group APS than in group CON for both transthoracic esophagectomy (22 +/- 20 versus 30 +/- 37, P = 0.005) and nontransthoracic esophagectomy patients (19 +/- 13 versus 25 +/- 21, P = 0.029). CONCLUSION: Adequate postoperative analgesia is associated with lower cardiopulmonary complications, lower mortality and reduced cost in patients undergoing transthoracic esophagectomy. PMID- 9207158 TI - Prospective, randomized evaluation of the efficacy of fibrin sealant as a topical hemostatic agent at the cannulation site in neonates undergoing extracorporeal membrane oxygenation. AB - BACKGROUND: The topical hemostatic effect of fibrin sealant that has been solvent/detergent treated and plasminogen depleted was evaluated in a multicenter prospective, randomized controlled study at the cannulation site wound of infants undergoing extracorporeal membrane oxygenation (ECMO). METHODS: The test group received standard cauterization and Fibrin sealant, while the control group was given cauterization alone to control hemostasis at this site. Efficacy data were available on 173 randomized study subjects of whom 149 met study entry criteria. All were managed according to standard ECMO practice. RESULTS: Fibrin sealant reduced the risk of bleeding, was associated with less shed blood, and was associated with shorter duration of hemorrhage. Further, control infants showed an increased bleeding risk with less depressed fibrinogen levels and prothrombin time elevations >18 seconds prior to ECMO. CONCLUSION: Fibrin sealant is useful as a topical hemostatic agent in patients with coagulopathy not responding to standard surgical techniques. PMID- 9207159 TI - The origin of lower extremity deep vein thrombi in acute venous thrombosis. AB - BACKGROUND: It has been taught that most deep venous thromboses (DVT) begin in the deep veins of the calf and propagate proximally. The duplex ultrasound scan, with its noninvasive characteristics and accuracy, has brought this premise into question. The purpose of this study was to determine the pattern and distribution of acute DVT as well as the different types of thrombi. METHODS: We performed a retrospective review of all duplex scans ordered for a diagnosis of acute lower extremite DVT at a 200-bed hospital over a 5-year period. RESULTS: There were 3,585 examinations performed on 2,654 patients. There were 461 patients (17.4%) with a venous thrombosis. Four types of venous thrombosis were identified: an isolated thrombosis in one venous segment (34%), a thrombosis extending over two or more contiguous segments (52%), multiple thromboses in noncontiguous segments (8%), and bilateral thrombi in different locations (6%). CONCLUSION: Calf vein thrombi represented 24% of all DVT. Thrombi in the major veins of the thigh and popliteal space without calf involvement were present in 49% of all DVT. The data in this paper indicate that most significant deep venous thromboses do not begin in the calf but instead arise in the proximal thigh or groin. PMID- 9207160 TI - Operative treatment of congenital stenoses of the intrahepatic bile ducts in patients with choledochal cysts. AB - BACKGROUND: Postoperative complications including intrahepatic calculi may develop after the complete excision of a choledochal cyst. Since congenital stenoses of the intrahepatic bile ducts are more likely the cause of intrahepatic calculi, operative procedures for intrahepatic stenoses are reported. METHODS: There were 16 patients with choledochal cysts who underwent surgery for stenoses of intrahepatic bile ducts. The stenoses were excised at the opening of the common hepatic duct. RESULTS: In the 16 patients, 25 of the 26 stenoses that involved an intraluminal membrane or septum could be excised from the divided end of the common hepatic duct at the hepatic hilum. In 1 patient, the stenosis could not be accessed from the hepatic hilum, and a left hepatic lobectomy was required. In postoperative follow-up, all 16 patients were in good health. CONCLUSIONS: Stenoses of the intrahepatic bile ducts should be treated from the divided end of the common hepatic duct at the initial operation for choledochal cysts. The need for a second operation or hepatic lobectomy may thus be avoided. PMID- 9207161 TI - Is incidental prophylactic oophorectomy an acceptable means to reduce the incidence of ovarian cancer? AB - BACKGROUND: According to previous reports, the lifetime risk of developing ovarian carcinoma is 1.4%. This figure varies with age from 6.6 per 100,000 among women aged 35 to 39 years up to 55.1 per 100,000 among women aged 75 to 79 years. Prophylactic oophorectomy remains a modality to decrease the incidence of ovarian cancer. What proportion of women diagnosed with an ovarian malignancy had a preceding laparotomy at which time a prophylactic oophorectomy could have been performed? METHODS: We reviewed the new ovarian cancer diagnoses seen in patients between August 1988 and August 1993 at the Ottawa Regional Cancer Foundation. Four hundred and four patients were identified. These patients were analyzed for preceding abdominal surgery, age, time to disease progression, time to death, time to death from other causes, and average follow-up. The previous abdominal surgeries were divided into: (1) major gynecological surgery; and (2) general surgery procedures, which were further divided into laparotomy and pelvic surgery (group A surgeries) and general surgery that included other abdominal surgeries (ie, appendectomy, cholecystectomy) where access to the pelvis could be more difficult (group B surgeries). RESULTS: A total of 270 abdominal surgeries was performed, prior to the diagnosis of ovarian cancer. The group was stratified according to the timing of the surgery (< or =40 years, 41 to 45 years, 46 to 50 years, >50 years). Based on these data, and on the grouping of general gynecologic surgeries plus the general surgical procedures of group A, 10.9% of ovarian cancers would have been prevented if prophylactic oophorectomy had been performed in patients who had surgery over 40 years of age; over 45 years this was 6.7%, over 50 years it was 4%. If one adds all major surgeries, including general surgery groups A and B, the results were 26.9% over 40 years of age, 20% over 45, and 16.6% over 50. CONCLUSION: We found that, depending on the age of the patient, prophylactic oophorectomy results in a 4% to 10.9% reduction in the incidence of ovarian carcinoma. This increases to 16.6% to 26.9% if one considers general surgery procedures in which access could be more difficult. Although we are not advocating the frequent use of this procedure, we recommend that surgeons routinely discuss this option before surgery with their postmenopausal female patients over 49 years of age. Given that the decision for prophylactic oophorectomy is multifaceted, we feel that a risk scoring for ovarian cancer and a discussion of the risk and benefit ratio should be undertaken. The ultimate goal is to heighten patient awareness of the risk factors to ensure that an informed decision is made concerning this consistently lethal disease. PMID- 9207163 TI - Lower extremity iatrogenic nerve injury due to compression during intraabdominal surgery. AB - BACKGROUND: latrogenic nerve injury due to poor positioning and external compression is a common surgical complication. However, sciatic neuropathy from external compression and femoral nerve injury after self-retaining retraction are less-published complications. METHODS: Surgical Morbidity and Mortality Reports from 1986 through 1995 were reviewed to identify femoral and sciatic neuropathies following intraabdominal vascular and general surgeries. RESULTS: Two sciatic and 5 femoral neuropathies were reported, an incidence of approximately 0.17% of abdominal cases. Sciatic injuries were attributed to external compression, whereas femoral neuropathies were due to compression by self-retaining retraction. The 3 female and 4 male patients had a mean age of 53.4 years, and no patient had a prior history of peripheral neuropathy. Mean operating time for sciatic injuries was 8.2 hours, versus 4.3 hours for femoral neuropathies. Both patients with sciatic neuropathy had complete resolution of symptoms, compared with 1 femoral neuropathy patient. Two femoral neuropathies were permanent, 1 had partial resolution and 1 had improvement at 4 months but was lost to follow-up. CONCLUSIONS: Sciatic and femoral compression neuropathies are rare but serious complications of abdominal surgery. When retracting in the deep pelvis, consideration should be given to using small, well-padded retractor blades and repositioning these regularly. Prevention of sciatic nerve compression requires careful padding of the table surface, especially for longer cases. PMID- 9207162 TI - Outpatient thyroidectomy. AB - In current clinical practice, the concept of outpatient surgery could apply to thyroidectomy. As the thyroid is anatomically accessible, its removal is not physiologically disabling; it makes surgery safer and precludes hospitalization. To evaluate the feasibility and solidity of outpatient thyroidectomy (OPT), the authors conducted a 12 1/2-year study (1982-1994), including an earlier 4-year randomized trial on 309 and cumulative posttrial experiences in 869 cases. The results showed the safety, practicality, and efficacy of OPT as compared with standard thyroidectomy. The study confirms the validity of OPT and is suggested for selected patients with thyroid disease. PMID- 9207164 TI - Description of new "bowel-sparing" techniques for long strictures of Crohn's disease. AB - In the period of January 1993 to December 1995 we operated on 55 patients with various complications of Crohn's disease. In properly selected cases, obstructive complications of Crohn's disease can be treated effectively by strictureplasty. Long strictures, even if a narrow lumen is still present, are commonly managed by resection, as classic strictureplasties cannot be done; also Finney strictureplasty seems inadequate, as it creates a blind loop that favors bacterial overgrowth and fecal stasis. Three original "sparing bowel" surgical approaches are proposed as possible alternative in the treatment of long stricture in Crohn's disease. We perform side-to-side ileoileal plasty whenever we are faced with severe narrowing of a long segment of small bowel (>10 cm); side-to-side ileocolic plasty whenever very severe disease with narrowing of ileocaecal valve is present; and ileocaecal plasty when terminal ileitis involves the very distal end of the small bowel, but sparing or only minimally affecting the ileocaecal valve. The above-mentioned procedures are described in detail and the clinical outcomes related to the first 8-patient series of our institution are presented. PMID- 9207165 TI - Early decision for conversion of laparoscopic to open cholecystectomy for treatment of acute cholecystitis. AB - BACKGROUND: Although recent reports suggest an initial laparoscopic approach to acute cholecystitis, the risk factors and consequences of the failure of an attempt remained unknown. METHODS: A retrospective study of 557 laparoscopic cholecystectomies was undertaken to identify 70 patients (13%) with a clinical diagnosis of acute cholecystitis confirmed by ultrasonography. Patients who required conversion to laparotomy (conversion group) were compared to those with successful laparoscopic cholecystectomy (successful group). RESULTS: Eight of 70 patients (11%) required conversion. The conversion group had significantly more elderly (< or =65 years) patients (88% vs 37%; P = 0.02) and larger gallstones as shown on ultrasonography (25 mm vs 15.5 mm; P = 0.03). Other preoperative factors associated with severe inflammation were not predictive. Conversion was associated with the intraoperative finding of severe adhesions and not with those of empyema of gallbladder or gangrenous cholecystitis. Conversion was made after a median laparoscopic surgery time of 50 minutes. The conversion group required more operation time, more analgesics, a longer recovery time, and a longer hospital stay. In addition, the postoperative complication rate was significantly higher (63% vs 16%; P = 0.009). CONCLUSIONS: Patients who required conversion from laparoscopic to open cholecystectomy for acute cholecystitis are at risk for postoperative complications. In elderly patients with large gallstones, the surgeon should made an early decision to convert if severe adhesions are encountered. PMID- 9207166 TI - Pancreatic endocrine tumors with loss of heterozygosity at the multiple endocrine neoplasia type I locus. AB - BACKGROUND: Loss of heterozygosity (LOH) at chromosome 11q13 has been demonstrated in multiple endocrine neoplasia type I (MEN I) and sporadic parathyroid tumors, pituitary adenomas, and a few types of pancreatic endocrine tumors. Gastrinomas are the most common pancreatic endocrine tumor in MEN I. We hypothesized that all pancreatic endocrine tumors have LOH at 11q13, resulting in inactivation of the previously described tumor suppressor gene in this region. METHODS: We analyzed a sporadic gastrinoma, a MEN I-associated gastrinoma, and a nonfunctional pancreatic endocrine tumor from a patient with Von Hippel-Lindau (VHL) disease for LOH at seven loci at 11q13: D11S149, PYGM, D11S427, D11S546, SEA, D11S97, and D11S146. RESULTS AND CONCLUSIONS: We found LOH at 11q13 in all three tumors. The MEN I-associated gastrinoma we analyzed is the first tumor of this type to have LOH. This is also the first report of LOH at 11q13 in a pancreatic endocrine tumor from a patient with VHL. These findings suggest that the etiology of pancreatic endocrine tumor formation involves a common genetic pathway for sporadic, MEN I, and VHL tumors. PMID- 9207167 TI - Loss of heterozygosity and homozygous deletion of the tpr locus in human gastric cancer. AB - BACKGROUND: Despite being one of the world's most common neoplasias, there is little information on the molecular events that lead to gastric cancer. Molecular studies have shown that inactivation of tumor suppressor genes by mutation and/or allelic loss is an important genetic alteration in the multistep process of tumorigenesis. METHODS: In an attempt to identify a putative tumor suppressor gene involved in the carcinogenesis of gastric cancer, we performed Southern blot analysis using the tpr probe for 44 patients with gastric cancer, using tumor tissue and normal tissue from the same specimen. RESULTS: Of the 44 samples, 7 (16%) were informative, heterozygous cases for the tpr probe. Three of the informative cases showed a loss of heterozygosity and 3 cases showed homozygous deletion for the tpr probe (6 of 7; 85%). CONCLUSIONS: These findings suggest that tpr gene plays a role in gastric tumorigenesis, and this may be due to a tumor suppressor effect for the tpr gene. PMID- 9207168 TI - The past, present, and future of lung transplantation. AB - BACKGROUND: The history of lung transplantation from the first human transplant performed in 1963 to the present is reviewed with particular focus on the added challenges because of the contaminated bronchus, exposure of the graft to airborne organisms, the poor blood supply to the bronchus, and the problem of reperfusion pulmonary edema. METHODS: The technical aspects of single and double sequential lung transplantation are reviewed, as are the current indications for single, double sequential, and heart/lung transplantation. Criteria for lung transplant recipients, in addition to their primary disease are noted, as are absolute and relative contraindications. The standard criteria for donor selection are also reviewed. RESULTS: The results of single, double sequential, and heart-lung transplantation over the past 10 years as reported by the International Society for Heart and Lung Transplantation Database are reviewed. In addition, the statistics of the lung and heart-lung transplantation program at the University of Colorado Health Sciences Center are reviewed, including the current immunosuppressive regimens and early and late monitoring for infection and rejection. This experience includes 3 early deaths in the first 53 patients for an operative mortality of 5.6%, with a 1-year actuarial survival of 90%. CONCLUSIONS: During the past decade remarkable improvement in the result of single and double sequential lung transplantation have occurred. As 1-year, actuarial survival is now approaching 90% at some institutions. Living related lobar transplantation, new antirejection agents, chimerism, and xenograft transplantation are areas for continuing and future investigation. The shortage in donor organ supply continues to be a very significant factor in limiting human lung transplantation. PMID- 9207169 TI - A time to listen. AB - As methods of health care delivery, and advances in technology change our lives and practices, an essential element of personal and professional relationships has become neglected. We have stopped listening to each other, to our patients, and to ourselves; we have lost the art of communication. The essential aspects of optimal communication and the power of nonverbal signals are reviewed. Only by recognizing the importance of communication in surgical education, practice, and in fact in all aspects of daily life, will this encroaching societal deafness be rebuffed. PMID- 9207170 TI - Running an objective structured clinical examination on a shoestring budget. AB - BACKGROUND: A major obstacle in the wide implementation of objective clinical structured examinations (OSCEs) is their high cost, averaging $200 to $300 (Canadian dollars, CDN) per candidate and estimated at up to CDN$900 per candidate if all "hidden" administrative costs are included. METHODS: A detailed cost analysis of preparing and administering OSCEs at 1 institution was undertaken over 2 years. An 18-station, 6-minute-per-station OSCE was given to a 72-student third-year medical class. RESULTS: The total cost of the OSCE was CDN$5,010, or $70 per student. The key factors in reaching this significantly lower per-student OSCE cost included: judicious use of standardized patients, use of academic faculty for preparing and marking the stations, and decreased secretarial and other administrative costs. CONCLUSIONS: Data suggest that OSCEs can be set up with reasonable cost and limited resources even in smaller institutions. Cost should not be considered a major obstacle in implementing this excellent examination type in undergraduate medical education. PMID- 9207171 TI - Teaching three-dimensional surgical concepts of inguinal hernia in a time effective manner using a two-dimensional paper-cut. AB - BACKGROUND: Because inguinal hernia repair is difficult for third-year students to comprehend, a 2-dimensional paper-cut was developed to teach the concepts of inguinal hernia in a time-effective manner before students' observation of herniorrhaphy in the operating room. METHODS: Using Adobe Illustrator 5.5 for MacIntosh, a 2-dimensional inexpensively printed paper-cut was created to allow students to perform their own simulated hernia repair before observing surgery. The exercise was performed using a no.15 scalpel or an iris scissors and was evaluated by comparing 10-question pre-tests and post-tests. RESULTS: Seventy five students performed the exercise, most completing it within 15 minutes. The mean pre-test score was 7.4/10 and the mean post-test score was 9.1/10. Students performing the paper-cut reported better understanding when observing actual herniorrhaphy. CONCLUSIONS: A 2-dimensional paper-cut ("surgical origami") may be a time-effective method to prepare students for the observation of hernia repair. PMID- 9207172 TI - Increasing examiner involvement in an objective structured clinical examination by integrating a structured oral examination. AB - INTRODUCTION: The role of physician examiners in an objective structured clinical examination (OSCE) is relatively passive. In our institution examiners criticized the passive nature of their role. This study evaluates the reliability and viability of adding a structured oral examination to an OSCE. METHOD: Ten 24 minute stations consisted of three parts. Part I: 12 minutes-patient encounter. Part II: 6 minutes-oral presentation covering findings, differential diagnosis, and management plan. Part III: 6 minutes-structural oral examination (SOE), containing 5 predetermined questions. RESULTS: Over 6 consecutive days, 72 graduates were assessed. Overall average score: 72.02 (SD 5.05); reliability 0.84. Part I of the OSCE average score: 69.2 (SD 7.4); reliability 0.69. Part II oral presentation average score 64 (SD 5.8) reliability 0.87. SOE average score 77.7 (SD 6.3); reliability 0.64. Eighty-nine percent of the examiners indicated satisfaction with the new format. CONCLUSIONS: The SOE was a reliable component of an OSCE and contributed to the overall reliability. Examiners reported a higher degree of satisfaction with the examination. PMID- 9207173 TI - Dual action of cAMP-dependent protein kinase on granulocyte movement. AB - Cell locomotion is a continuous cycle of integrin-dependent attachments and detachments along chemotactic gradients, driven by dynamic modulations of the actin network. Cyclic AMP (cAMP), which is known to be generated by N-formyl-l methionyl-l-leucyl-l-phenylalanine (fMLP) receptors but not by beta2 integrins, was investigated as a coordinator of granulocyte locomotion. Elevation of cAMP by exposure to forskolin (100 microM) and 1-isobutyl-methylxanthine (IBMX; 100 microM) caused a marked reduction in beta2 integrin-induced polymerisation of actin, but had a less pronounced effect on the fMLP-induced actin response. Pretreatment of cells with rp-adenosine-3',5'-cyclic monophosphorothioate (rp cAMPS; 50 microM), an inhibitor of the cAMP-dependent protein kinase (cAPK), resulted in a significant increase in the fMLP-induced actin polymerisation response. In agreement with the effect on filamentous actin (F-actin) forskolin and IBMX markedly suppressed the migration of granulocytes towards fMLP. Surprisingly enough, pretreatment of cells with rp-cAMPS inhibited cell movement to the same extent as forskolin and IBMX did. This dual action of cAMP on granulocyte migration suggest an important regulatory mechanism whereby the balance of this intracellular signal results in an optimal locomotory response. PMID- 9207174 TI - Effects of 4-OH-2,3-trans-nonenal on human erythrocyte plasma membrane Ca2+ pump and passive Ca2+ permeability. AB - Structural and functional alterations of cell membranes caused by free radicals leading to lipid peroxidation and increases in intracellular Ca2+ concentrations have been implicated in atherogenesis. The objective of this study was to directly measure the effects of a major aldehydic lipid peroxidation product, 4 OH-nonenal (HNE), on plasma membrane Ca2+ regulatory mechanisms. This was attained by measuring passive Ca2+ permeability, primary active Ca2+-transport, and (Ca2+ + Mg2+)-ATPase activity in a human red cell model system. Using this three-pronged approach it could be shown that HNE increases passive Ca2+ permeability significantly beyond the typically low basal flux, while at the same time inhibiting the active Ca2+ extrusion pump and associated (Ca2+ + Mg2+) ATPase activity. We conclude that this deleterious combination of effects by HNE in this plasma membrane model system may be indicative of plasma membrane changes in cells directly involved in atheroma formation and thus may represent causative factors in the early stages of atherogenesis. PMID- 9207175 TI - Association of coatomer proteins with the beta-receptor for platelet-derived growth factor. AB - The nonreceptor tyrosine kinase Src binds to and is activated by the beta receptor for platelet-derived growth factor (PDGF). The interaction leads to Src phosphorylation of Tyr934 in the kinase domain of the receptor. In the course of the functional characterization of this phosphorylation, we noticed that components of 136 and 97 kDa bound to a peptide from this region of the receptor in a phosphorylation-independent manner. These components have now been purified and identified as alpha- and beta'-coatomer proteins (COPs), respectively. COPs are a family of proteins involved in the regulation of intracellular vesicle transport. In order to explore the functional significance of the interaction between alpha- and beta'-COP and the PDGF receptor, a receptor mutant was made in which the conserved histidine residue 928 was mutated to an alanine residue. The mutant receptor, which was unable to bind alpha- or beta'-COP, showed a normal ligand-induced autophosphorylation. The mutant receptor also behaved like the wildtype receptor with regard to biosynthesis and maturation, and mediated a mitogenic signal. The possible functional importance of the interaction between the PDGF beta-receptor and alpha- and beta'-COP, is discussed. PMID- 9207176 TI - The role of phorbol ester-sensitive protein kinase C isoforms in lymphokine activated killer cell-mediated cytotoxicity: dissociation between perforin dependent and Fas-dependent cytotoxicity. AB - Treatment of lymphokine-activated killer (LAK) cells with phorbol ester (PMA) caused the downmodulation of LAK activity concomitantly with the inhibition of serine esterase (SE) release, which has been shown as a marker for perforin dependent cell-mediated cytotoxicity. The reduction of perforin-dependent LAK activity by PMA-treatment appeared to be due to the disappearance of PMA sensitive protein kinase C (PKC) isoforms such as PKC alpha, gamma, epsilon, theta. In contrast, Fas-mediated LAK activity was refractory against PMA-induced downregulation. Treatment of LAK cells with PMA caused a disappearance of cytotoxicity against Fas L5178Y tumor cells, while cytotoxicity against Fas+ transfectants was not affected by PMA treatment. Moreover, Fas-mediated LAK activity of perforin-knockout mice was not inhibited by PMA treatment. These results clearly demonstrated that Fas-mediated cytotoxicity could be dissociated from perforin-mediated cytotoxicity by their different requirement of PMA sensitive PKC isoforms. PMID- 9207177 TI - Cytosolic Ca2+ gradients in pancreatic islet-cells stimulated by glucose and carbachol. AB - Digital image analysis was employed to resolve the spatial differences in distribution of cytosolic free Ca2+ concentrations ([Ca2+]i) in mouse pancreatic islet-cells stimulated with glucose and carbachol. Using Indo-1 loaded mouse islet-cells, we have demonstrated that glucose induces steep spatial gradients of [Ca2+]i in isolated mouse islet-cells. Furthermore, the largest [Ca2+]i increase was always spatially restricted to a region just beneath the plasma membrane. Low concentrations of carbachol (0.6 microM) induced steep spatial gradients of [Ca2+]i which originated from the center of the cells. However, 10 microM carbachol increased [Ca2+]i to high levels collapsing the [Ca2+]i gradients in the center of the cells. Different patterns of [Ca2+]i oscillations were observed between dissociated pancreatic islet-cells and mouse pancreatic islets when challenged with 11 mM glucose. Under these conditions we could identify cells within the islet which oscillate with the same pattern as the whole islet. We postulate that "initiators" of insulin release, as glucose, induce greater [Ca2+]i increases at exocytotic sites than those induced by "potentiators", as carbachol. PMID- 9207178 TI - Protonophoric activity of NADH coenzyme Q reductase and ATP synthase in coupled submitochondrial particles from horse platelets. AB - A method to prepare coupled submitochondrial particles from horse platelets is described. The method allowed us to study the protonophoric activities of both complex I and complex V following the fluorescence quenching of the monoamine 9 amino-6-chloro-2 methoxyacridine (ACMA), a probe highly sensitive to the generation of a transmembrane delta pH. We carried out a kinetic analysis of each enzyme complex studying the proton translocation and the electron transfer activities of complex I as well as the proton translocation and the ATP hydrolytic activities of complex V. A micromethod to prepare coupled submitochondrial particles from platelets might be useful to investigate cell bioenergetic damage occurring in mitochondrial diseases and ageing. PMID- 9207179 TI - Expression of ST2, an interleukin-1 receptor homologue, is induced by proinflammatory stimuli. AB - ST2/T1 is an orphan receptor highly homologous to the IL-1 receptor. Using ST2 cDNA, ST2 specific primers, and a polyclonal antibody generated against ST2, the expression of mRNA and protein corresponding to both the soluble and membrane anchored forms of ST2 was studied. ST2 mRNAs were ubiquitously expressed in all the human tissues examined and were induced by cytokines and phorbol esters. Three different species of mRNAs were observed in different human cells and tissues. In contrast, only two species of ST2 mRNAs were observed in murine Balb/c-3T3 cells and no ST2 mRNA was seen in most tissues of normal mice. However, in a murine model where mouse ears are exposed to UVB irradiation leading to inflammation, ST2 mRNA was expressed 48 h post UV exposure. Similarly, in Balb/c-3T3 cells, the expression of soluble ST2 mRNA and protein was induced by pro-inflammatory stimuli such as TNF, IL-1alpha, IL-1beta and PMA in both exponentially growing and quiescent cells. The expression of the membrane ST2, however, remained constant. These data suggest a role for ST2 in inflammation. PMID- 9207180 TI - A putative tetrapeptide antagonist prevents beta-amyloid-induced long-term elevation of [Ca2+]i in rat astrocytes. AB - Comparative fluorimetric studies on the long-term (8-hour) action of beta[1 42]amyloid and its shorter fragments beta[1-40], beta[25-35] and beta[31-35] on the steady-state intracellular Ca2+ concentration in primary cultures of rat astroglial cells using the Ca2+-sensitive fluorescent probe Fura-2 AM revealed higher 340/380 fluorescence excitation ratios in the treated cells as compared to the untreated controls. All the peptides were found to induce similar cellular effects, suggesting the [31-35] region as the putative active centre of the molecule. No significant alteration was detectable in Fura-2 fluorescence using the Ca2+-insensitive excitation wavelength of 367 nm, indicating that the observed changes reflect a real alteration in the Ca2+ concentration of the cells. Moreover, no considerable difference was observed in the total protein content of treated and untreated cells. Co-treatment of the cells with Pr-Ile-Ile Gly-Leu-NH2 (Pr-IIGL) peptide, an analogue of the [31-34] region of beta[1-42] amyloid, was found to effectively antagonize the beta[1-42]-amyloid-induced elevation of the fluorescence excitation ratio, leaving the 367-nm fluorescence unaffected. To the best of the authors' knowledge, this is the first report on an analogue of beta-amyloid peptide capable of blocking one of its physiological effects, thereby raising the possibility that this sequence could prove to be a lead compound for designing effective beta-amyloid antagonists. PMID- 9207181 TI - Endogenous ADP-ribosylation of a G(alpha i) protein in bovine ciliary body is stimulated by nitric oxide. AB - Five ciliary body membrane proteins were labeled when incubated with (adenylate 32P)NAD. Nitric oxide donors stimulated the labeling of 64, 40, and 29-30 kDa proteins and inhibited that of 58 and 56 kDa proteins. The greatest influence of nitric oxide was on the 40 kDa protein: a 17-fold stimulation. Western blotting and immunoprecipitation with specific antibodies identified this protein as the alpha-subunit of G(i-1). Studies with inhibitors showed that the protein was mono ADP-ribosylated. Treatment of (32P)NAD-labeled G(i-1) with Hg and analysis of the released radioactive material showed that the protein was ADP-ribosylated on a cysteine residue. PMID- 9207182 TI - Expression of a kinase-defective Eph-like receptor in the normal human brain. AB - We have identified a human Eph-family protein, HEP, gene located in human chromosomal region 7q33-->q35. The deduced amino acid sequence shared primary structural properties of Eph-family receptor tyrosine kinases. However, six invariant amino acids such as a lysine in the ATP-binding site and an aspartic acid in the phosphotransfer site of a conserved catalytic domain were substituted with other amino acid residues in HEP. Thus, no intrinsic tyrosine kinase activity was detectable in the catalytic domain expressed in CHO-K1 cell transfectants. Although most kinase-defective mutants of growth factor receptors have been reported as pathogenic receptors, its transcript was abundantly expressed in normal human adult tissues. A 135-kDa HEP protein was expressed in the human brain as much as in CHO-K1 cells transfected with a HEP cDNA expression vector. HEP is the first description of a kinase-defective Eph-family protein expressed abundantly in normal human tissues. PMID- 9207183 TI - Placenta growth factor: identification and characterization of a novel isoform generated by RNA alternative splicing. AB - We report the isolation and characterization of a third isoform of placenta growth factor (PlGF), generated by alternative splicing of its RNA transcript. This novel form of PlGF, PlGF-3, was cloned by the polymerase chain reaction technique using a human cDNA library prepared from the terminal placental tissue. PlGF-3 contains an in-frame insertion of 72-amino acids near the C-terminal portion of PlGF-1. Southern blot analysis revealed that a single gene encoded PlGF-2 and PlGF-3. Nucleic acid sequence analysis found the insertion of 216 nucleotides of PlGF-3 between exon 4 and exon 5 of the PlGF gene. Northern blot and tissue distribution studies discovered two mRNA species for PlGF-3, which were both uniquely expressed in the placenta. Transient expression of PlGF-3 cDNA in mammalian cells showed PlGF-3 was detected in the conditioned medium as both dimers and monomers. Unlike PlGF-2, PlGF-3 lacked heparin-binding affinity. Thus, alternative splicing of PlGF RNA produces at least three polypeptides with different secretion pattern, heparin-binding affinity and dimerization properties. PMID- 9207184 TI - cDNA cloning and genomic organization of the mouse BMP type II receptor. AB - The cDNA for the mouse bone morphogenetic protein type II receptor (BMPR-II) was isolated using the human counterpart as a probe and its genomic structure was determined. The cDNA encodes a protein of 1,038 amino acids with a single transmembrane domain, a serine/threonine kinase domain, and a long carboxy terminal tail. The overall amino acid sequence identity between the mouse and the human BMPR-II is 96.6%. mRNA is widely distributed in various adult tissues. The gene is encoded by 13 exons spanning over 80 kb. Two large introns (intron 1 and 3) contribute to the majority of the gene size, as in the mouse activin type II receptor gene. The intron/exon boundaries were sequenced. The results suggest that alternative splicing can yield a shorter form of BMPR-II of 530 amino acids, as reported previously. Knowledge of the structure of the BMPR-II gene is essential for the understanding of the role of bone morphogenetic proteins in the developmental and physiological processes of animals. PMID- 9207185 TI - Hepatitis G virus replication in human cultured cells displaying susceptibility to hepatitis C virus infection. AB - We recently developed a hepatitis C virus (HCV) replication system using two cultured human cell lines: MT-2C, a human T-cell leukemia virus type I-infected cloned T cell line, and PH5CH, a non-neoplastic human hepatocyte line. In the course of the study, we found evidence that replication of hepatitis G virus (HGV), which has recently been identified, was supported in both MT-2C and PH5CH cells. When these cells were inoculated with serum 1B-3 containing both HCV and HGV obtained from a blood donor, the 5'-noncoding (5'-NC) regions of HCV RNA and HGV RNA were detected by RT-nested PCR in both cell lines more than 30 days postinoculation, and the 3'-noncoding region of HGV RNA was also detected in the both cell lines more than 30 days postinoculation. Sequence analysis of the 5'-NC region of HGV RNA revealed the quasispecies nature of HGV, although the 5'-NC region was highly conserved among HGV isolates. This HGV-infected culture system will be useful for various biological and virological studies of HGV. PMID- 9207186 TI - Flavone acetic acid stimulates nitric oxide and peroxynitrite production in subcutaneous mouse tumors. AB - Flavone acetic acid (FAA) has powerful anti-tumor activity against many types of solid murine tumors, but its biochemical mechanism of action is not understood. The present study examined the role of tumor vasculature and nitric oxide in mediating the anti-tumor effects of FAA. Athymic nude mice bearing subcutaneous RJ2-14 tumors were treated with a single dose of FAA, 200 mg/kg i.p., and euthanized at various times. Apoptosis within tumors was apparent during the first six hours of FAA treatment. We found that Type III, endothelial nitric oxide synthase (NOS) activity was significantly increased in tumors, but not in other tissues, as early as two hours after FAA dosing. FAA also stimulated the formation of the toxic peroxynitrite radical in tumors within two hours of treatment as assessed by immunostaining for nitrotyrosine. Staining was observed in dilated tumor vessels and surrounding tumor cells and correlated with the presence of apoptosis. Tumor endothelium may therefore be a critical target for FAA activity via stimulation of the nitric oxide pathway. PMID- 9207187 TI - Lead discovery of inhibitors of the dihydrofolate reductase domain of Plasmodium falciparum dihydrofolate reductase-thymidylate synthase. AB - A three-dimensional structure model of the dihydrofolate reductase (DHFR) domain of the bifunctional DHFR-thymidylate synthase of Plasmodium falciparum was used as a basis for computational screening of commercially available compounds for candidate inhibitors. Compounds which can stably dock to the model with strong ionic hydrogen bonds via protonation by an aspartic acid residue at the bottom of the active site were identified through docking simulation. Among compounds thus identified, 21 were assayed for inhibitory activity towards the recombinant DHFR domain. Two compounds, 2-amino-1,4-dihydro-4,4,7,8-tetramethyl-s-triazino(1,2 a)benzimida zole and Trp-P-2, inhibited the recombinant P. falciparum DHFR domain with Ki values of 0.54 and 8.7 microM, respectively. Kinetic analysis showed that these compounds competitively inhibited the enzyme with respect to the substrate dihydrofolate. These findings support the validity of both the modeled structure and the docking results. Furthermore, these compounds serve as leads for developing new DHFR inhibitors, since their skeletal structures are different from any of known DHFR inhibitors. This paper also reveals a new biological activity of Trp-P-2, a potent mutagen. PMID- 9207188 TI - Induction of heparin binding epidermal growth factor-like growth factor and amphiregulin mRNAs by gastrin in the rat stomach. AB - The present study was designed to investigate whether heparin-binding epidermal growth factor-like growth factor and its related peptides are expressed in response to gastrin in rat stomach. Rat gastrin-17I (2.5 nmol/kg/hour) or gastrin 17I plus gastrin receptor antagonist, L-740,093 (2.0 mg/kg/hour), was injected intravenously into male Sprague-Dawley rats. RNA was extracted from oxyntic mucosa, and heparin-binding epidermal growth factor-like growth factor and related peptide gene expression was estimated using a ribonuclease protection assay. The level of transforming growth factor-alpha mRNA did not change at any time point during the experiment. In contrast, the levels of heparin-binding epidermal growth factor-like growth factor and amphiregulin mRNA were significantly increased within 3 hours following gastrin infusion and reached maximum levels 6 and 12 hours later, respectively. Continuous infusion of gastrin significantly increased oxyntic mucosal proliferation. Gastrin receptor antagonist significantly inhibited gastrin-induced heparin-binding epidermal growth factor-like growth factor and amphiregulin gene expression and gastrin induced oxyntic mucosal proliferation. These findings indicate that heparin binding epidermal growth factor-like growth factor and amphiregulin genes are induced by gastrin and that they play a role in the trophic action of gastrin on oxyntic mucosa. PMID- 9207189 TI - Integrin alpha6 is involved in follicular growth in mice. AB - We previously reported that integrin alpha6 is expressed on granulosa cells in the inner layers of the human and porcine ovarian follicles, where granulosa cells have no direct contact with basal lamina. In this study, we examined the physiological role of integrin alpha6 on follicular growth with an immature superovulated mice model using the anti-integrin alpha6 monoclonal antibody, GoH3. In the ovaries of 9- to 20-day-old mice, integrin alpha6 was detected on all the layers of granulosa cells in the primordial, primary, and secondary follicles by immunohistochemistry. The 13-day-old female mice were superovulated by pregnant mare serum gonadotropin and human chorionic gonadotropin with the treatment of intraperitoneal administration of GoH3, or a control antibody, or PBS. In the group of GoH3 treatment, successful ovulation was observed in 57+/ 25.7% of the animals, whereas no ovulation was observed in the control groups (p<0.01). These findings indicate that integrin alpha6 is involved in gonadotropin-induced follicular growth. PMID- 9207190 TI - Expression of recombinant rat interleukin-13 (IL-13) and generation of a neutralizing rat IL-13 antiserum. AB - Using baculoviral and bacterial systems, we expressed biologically active recombinant rat IL-13 and generated neutralizing rat IL-13 antiserum. Recombinant rat IL-13 produced by baculovirus-infected insect cells stimulated proliferation of TF-1 premyeloid cell line and induced expression of 15-lipoxygenase mRNA in human peripheral blood monocytes. Antiserum generated by immunizing a rabbit with recombinant bacterial rat IL-13 specifically inhibited TF-1 proliferation induced by baculoviral rat IL-13 but did not neutralize human IL-13 mitogenic activity. Western blotting with anti-rat IL-13 serum revealed a approximately 12 kD protein band in supernatants of insect cells infected with recombinant baculovirus carrying the rat IL-13 cDNA. The availability of recombinant rat IL-13 and rat IL 13 antibodies should facilitate studying the role of IL-13 in rat models of human inflammatory disorders. PMID- 9207191 TI - Novel homologues of CSBP/p38 MAP kinase: activation, substrate specificity and sensitivity to inhibition by pyridinyl imidazoles. AB - A novel homologue of p38 MAP kinase, called SAPK4, has been cloned which shares 61% amino acid identity with p38 and is expressed predominantly in testes, pancreas and small intestine. We also cloned an alternative form of p38beta, termed p38beta2, which lacks the additional 8 amino acid insertion unique to p38beta. p38, p38beta, p38beta2, ERK6/p38gamma/SAPK3, and SAPK4 were characterized with respect to stimulus-dependent activation in transfected cells, substrate specificity, and sensitivity to inhibition by pyridinyl imidazoles. All homologues were stimulated, although to differing extents, by IL-1beta, TNF, sorbitol, and UV. Only SAPK3 and SAPK4 were stimulated significantly by PMA. p38beta showed the weakest activation overall. MBP, ATF-2, and both MAPKAP kinase 2 and kinase-3 were good substrates of p38 and p38beta in vitro. In contrast, only MBP, ATF2, and MAPKAP kinase-3 proved to be significant substrates of SAPK3 and SAPK4, and of these three, MAPKAP kinase-3 was by far the weakest. p38beta had very poor kinase activity for all substrates except MBP. While both p38 and p38beta2 were comparably inhibited by SB 203580 and SB 202190, neither SAPK3 nor SAPK4 were inhibited. p38beta was partially inhibited by both inhibitors. These data suggest that SAPK3 and SAPK4 form a distinct subset of the p38 MAP kinases with different expression pattern, response to stimuli, substrate specificity, and inhibitor sensitivity. PMID- 9207192 TI - Induction of apoptosis and altered nuclear/cytoplasmic distribution of the androgen receptor and prostate-specific antigen by 1alpha,25-dihydroxyvitamin D3 in androgen-responsive LNCaP cells. AB - In addition to suppressing prostate cell growth, vitamin D also up-regulates the expression of androgen receptor (AR) and prostate-specific antigen (PSA). To study the mechanism involved in the control of these proteins, LNCaP cells were treated with 10 nM 1alpha,25-dihydroxyvitamin D3 and separated into cytosol and nuclear fractions. AR and PSA were analyzed by western blot analysis. A second approach involved incubating control and treated cells with [3H]R1881, fractionating the cells into the cytosolic and nuclear components, and quantifying the amount of radioactivity associated with the respective fractions. Alternatively, immunohistochemical assays were performed by staining cells with cognate antibodies for AR and PSA. Both biochemical and immunohistochemical analyses show proportionately greater increased presence of AR in the nucleus, accompanied by relatively reduced AR in the cytosol, following treatment of LNCaP cells with vitamin D3. Surprisingly, PSA was found to be present in the nuclear fraction in both control and treated cells. These results suggest that vitamin D3 promotes the translocation of AR from the cytosol to the nucleus. The presence of PSA in the nucleus of LNCaP cells raises the possibility of an autogenous mode of control of PSA gene expression. PMID- 9207193 TI - Purification and characterization of a homodimeric catalase-peroxidase from the cyanobacterium Anacystis nidulans. AB - Cytosolic extracts of the cyanobacterium Anacystis nidulans exhibit both catalase and o-dianisidine peroxidase activity. Native polyacrylamide gel electrophoresis demonstrates one distinct enzyme, which has been purified to essential homogeneity and found to be composed of two identical subunits of equal size (80.5 kDa). The isoelectric point is at pH 4.7. It is a very efficient catalase with a broad pH optimum between 6.5 and 7.5 and a Km for H2O2 of 4.3 mM, a calculated turnover number of 7200 s(-1), and an overall-rate constant of 3.5 x 10(6) M(-1) s(-1). The behaviour of this protoheme-enzyme is typical of the class of prokaryotic catalase-peroxidases, which is sensitive to cyanide (Ki = 27.2 microM) and insensitive to the eukaryotic catalase inhibitor 3-amino-1,2,4 triazole. The enzyme accepts electrons from o-dianisidine, but not from ascorbate, glutathione, and NADH. With hydrogen peroxide in steady-state conditions the enzyme is mainly in the ferric state indicating that Compound I is much faster reduced by H2O2 than it is formed. The native enzyme is in the high spin state, which is transformed to low-spin upon addition of cyanide. With peroxoacetic acid Compound I is formed at a rate of 5.9 x 10(4) M(-1) s(-1) at pH 7.0 and 25 degrees C with about 50% hypochromicity, a Soret-maximum at 405 nm and isosbestic points at 354 and 427 nm. PMID- 9207194 TI - Transient down-regulation of inhibin-betaC expression following partial hepatectomy. AB - Inhibin-betaC is a recently described TGF-beta family member most homologous to inhibin-betaA and inhibin-betaB. By Northern analysis, inhibin-betaC mRNA was detected exclusively in the liver among a large number of adult mouse tissues surveyed. The expression of inhibin-betaC mRNA in adult liver dropped sharply and transiently following partial hepatectomy. At 6 and 12 hours following partial hepatectomy, the levels of inhibin-betaC mRNA were at least 8-fold lower than in control animals. The liver specificity of inhibin-betaC expression and its down regulation following partial hepatectomy suggest that inhibin-betaC may function as a negative regulator of liver growth. PMID- 9207195 TI - Evidence for a tissue-specific induction of cutaneous CYP2E1 by dexamethasone. AB - We studied in mouse the effect of topical application of dexamethasone or salicylic acid, on CYP2E1 and CYP3A expression (proteins and/or mRNA) in liver and skin. Dexamethasone was also administered by intraperitoneal injection. Topical application or intraperitoneal injection of dexamethasone increased cutaneous CYP2E1 (8 and 4-fold respectively) whereas the hepatic level of this isoform showed a slight decrease and hepatic CYP3A expression was increased (3 fold). Cutaneous CYP2E1 was increased (3-fold) after topical treatment by salicylic acid. This compound had no effect on hepatic CYP3A and CYP2E1 expression. Cutaneous CYP3A (protein and mRNA) was not detectable in all groups (control or treated animals). Dexamethasone and salicylic acid increased cutaneous CYP2E1 mRNA level (2.5 and 1.4-fold respectively). In conclusion, dexamethasone and salicylic acid induced cutaneous CYP2E1 protein and mRNA level. Cutaneous CYP2E1 induction by dexamethasone is a tissue-specific process. PMID- 9207196 TI - Cleavage specificity of human rhinovirus-2 2A protease for peptide substrates. AB - Substrate requirements of the human rhinovirus serotype-2 2A protease have been examined using synthetic peptides. A chromogenic peptide with a sequence of TRPIITTA-p-nitroanilide was found to be cleaved efficiently by the 2A protease with an apparent Km value of 95 microM, which allowed the protease activity to be monitored and measured continuously using a spectrophotometer. Competition cleavage assays reveal this peptide was cleaved over 10-fold more efficiently than the 16-mer peptide derived directly from its native processing site. On the basis of these data, we conclude that the P1' glycine residue is not absolutely needed for the 2A cleavage to occur and the essential residues required for the 2A activity would exist within the N-terminal side of the scissile bond. PMID- 9207197 TI - The endogenous mu-opioid receptor agonists endomorphins 1 and 2 have novel hypotensive activity in the rabbit. AB - The endogenous peptides endomorphins 1 and 2 are newly isolated, potent, and selective mu-opioid receptor agonists. In the present study, responses to the endomorphin peptides were investigated in the systemic vascular bed of the rabbit. Endomorphins 1 and 2 induced dose-related decreases in systemic arterial pressure when injected in doses of 1-30 nmol/kg i.v. In terms of relative vasodepressor activity, endomorphins 1 and 2 were similar to the ORL1 receptor ligand, nociceptin (Orphanin FQ), and met-enkephalin in decreasing systemic arterial pressure. Vasodepressor responses to endomorphins 1 and 2 were inhibited by the opioid receptor antagonist, naloxone, in a dose of 2 mg/kg i.v. These results demonstrate that endomorphins 1 and 2 have significant naloxone sensitive, vasodepressor activity in the rabbit. PMID- 9207198 TI - DNA topoisomerase II as the cellular target of a novel antitumor agent ICRF-193, a bisdioxopiperazine derivative, in Xenopus egg extract. AB - We have investigated the molecular target of an antitumor agent ICRF-193, a bisdioxopiperazine derivative, in in vitro chromosome condensation system of Xenopus egg extract (XEE), where DNA topoisomerase II was previously demonstrated to play a crucial role. Demembranated Xenopus sperm head chromatin is converted to metaphase chromosome-like structure in XEE in two steps, i.e., swelling of the chromatin followed by condensation of chromosome. When ICRF-193 was added to the reaction, swelling of the chromatin was not affected but chromosome condensation was completely blocked. This blockade was reversed by exogenous supplement of calf thymus topoisomerase II, which was in turn neutralized by anti-topoisomerase II monoclonal antibody. These results demonstrate that topoisomerase II is the molecular target of the drug ICRF-193. PMID- 9207199 TI - Modulation of macrophage inflammatory protein-1alpha and its receptors in human B cell lines derived from patients with acquired immunodeficiency syndrome and Burkitt's lymphoma. AB - Macrophage inflammatory protein-1 alpha (MIP-1alpha) is a member of the -C-C- family of low-molecular weight chemokines. MIP-1alpha is involved in the homeostatic control of stem cell proliferation, in inducing chemotaxis, and also in inflammatory responses in mature cell types. In order to observe modulations of MIP-1alpha secretion and expression along with MIP-1alpha receptor (MIP-1alpha R) expression for a possible autocrine role in AIDS associated B-cell lines, we studied a wide panel of human B-cell lines. Previous work by us has shown that HIV-1 tat down modulates MIP-1alpha by inducing a novel transcription factor MNP. Our data in this report suggest that HIV down modulates MIP-1alpha as a mechanism to evade suppression by this chemokine in human B-cells. Furthermore, our results strongly suggest MIP-1alpha autocrine loops in a majority of tumor B-cells as evident by MIP-1alpha-R expression, and also secretion of MIP-1alpha. PMID- 9207200 TI - Characterization of the genomic structure and promoter of the mouse NAD+ dependent 15-hydroxyprostaglandin dehydrogenase gene. AB - The mouse NAD+-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH) gene and its 5'-flanking region was cloned from a 129 mouse ES bacteriophage P1 genomic library. The gene contains 7 exons and 6 introns and is 11.3 kb in length. The transcription initiation site was mapped at 35 bases upstream from the ATG start codon. The nucleotide sequence of the 1.6 kb promoter region contains two TATA boxes and a number of potential regulatory elements including Sp1, CRE, GRE, AP1, AP2, NF-IL6 and estrogen receptor binding site. Studies of the promoter's activity showed that the first 400 nucleotides of 5'-flanking region efficiently drove the transcription of the luciferase reporter gene in U936 cells upon stimulation with a phorbol ester. PMID- 9207201 TI - Prolactin-mediated inhibition of 20alpha-hydroxysteroid dehydrogenase gene expression and the tyrosine kinase system. AB - The rat luteal 20alpha-hydroxysteroid dehydrogenase plays a key role at catabolizing progesterone and at decreasing the level of this steroid secreted by the ovaries. Throughout pregnancy and before parturition neither the mRNA nor the protein for this enzyme could be detected. In this investigation we set to examine whether PRL and PRL-like hormone from placental origin silence the expression of this gene and whether PRL action involves tyrosine kinase activity and/or de novo protein synthesis. The results revealed that PRL and PRL-like hormone from rat placental origin (rPL-1 and rPL-2), but not rat growth hormone, caused a rapid and profound inhibition of 20alpha-HSD mRNA expression in highly luteinized granulosa cells. Immunoprecipition and western blot analysis indicate that PRL-R associates with JAK2 and Stat5, and this association is increased within 30 seconds with PRL treatment. Although both JAK2 and Stat5 were phosphorylated on tyrosine upon PRL treatment, the PRL mediated inhibition of 20alpha-HSD was not reversed by either tyrosine kinase inhibitors, AG18 and genistein, but was largely reversed by the protein synthesis inhibitor cycloheximide. In summary, results of this investigation indicate that although PRL can activate the JAK2/Stat5 system in the corpus luteum, the down regulation of 20alpha-HSD mRNA by PRL does not appear to involve tyrosine kinase activity but depends on de novo synthesis of protein(s). PMID- 9207202 TI - RNA helicase activity of Escherichia coli SecA protein. AB - SecA protein of Escherichia coli (E. coli), an ATPase essential for the translocation of precursor proteins, was found to have an additional activity of RNA helicase. This RNA unwinding activity of SecA was tested with two kinds of RNA duplex with different predicted stability. Each of these duplexes is consisted of two strands of unequal length with single-stranded ends. The RNA helicase activity of SecA required ATP and divalent cations. Confirmation of this activity came from the inhibition of unwinding of the RNA duplex when SecA was preincubated with its own polyclonal antibody. The biological significance of the RNA helicase activity of E. coli SecA protein is discussed. PMID- 9207203 TI - Renal cortical expression of mRNAs for antioxidant enzymes in normal and diabetic rats. AB - Increased oxidative stress has been implicated in the development of vascular complications of diabetes. In this study, we examined the hypothesis whether chronic hyperglycemia induces oxidative stress by lowering renal expression and activity of antioxidant enzymes and a decrease in glutathione, an antioxidant, in streptozotocin diabetic rats. The results show that the expression of mRNAs for Cu/Zn superoxide dismutase and glutathione peroxidase was significantly increased and that of catalase was decreased in diabetic rats. However, the superoxide dismutase activity was significantly lower in diabetic than normal glomeruli, whereas the activities of the other two enzymes correlated with their mRNA expression. Total glutathione content was significantly decreased in diabetic compared to normal glomeruli. The data suggest that hyperglycemia induces oxidative stress by overexpressing rather than lowering certain antioxidant enzyme mRNAs in the kidney of diabetic rats. Enhanced nonenzymatic glycation of enzyme protein seems to be the cause for the observed decrease in glomerular superoxide dismutase activity. PMID- 9207204 TI - beta-Galactosidase reporter system in mycobacteria and its application in rapid antimycobacterial drug screening. AB - Pathogenic mycobacteria are generally slow growing organisms and it takes several weeks to evaluate inhibitors of growth. Therefore, for rapid screening of the inhibitors of mycobacterial growth, a beta-galactosidase reporter system has been described which utilises a recombinant Mycobacterium smegmatis mc(2)155 expressing E. coli lacZ gene as the test organism. The assay is based on production of beta-galactosidase in presence of drugs during growth. A correlation between beta-galactosidase production and colony forming ability of mycobacteria was obtained. beta-galactosidase production was inhibited within 6 h by front line standard antimycobacterial drugs like streptomycin, rifampicin, isoniazid, ethambutol, pyrazinamide and ofloxacin at their reported MICs. The assay was performed on mycobacterial cells permeabilized with chloroform and sodium dodecyl sulfate. PMID- 9207205 TI - Trypsin isolated from the midgut of the tobacco hornworm, Manduca sexta, is inhibited by synthetic pro-peptides in vitro. AB - The seven residue synthetic peptides VPAYPQR and VPANPQR, identical to the pro regions of midgut trypsins from Manduca sexta, inhibit the peptidase activity of the mature enzymes. The inhibitory potencies of these peptides towards purified insect trypsin were determined and were found to be more potent at pH 7.0 than at pH 10.0. These peptides did not inhibit porcine trypsin, nor were they degraded rapidly by purified insect trypsin. We suggest that these particular peptides, and insect proregions in general, may be suitable for use in genetically engineered plants as insect specific inhibitors of digestive proteases. PMID- 9207206 TI - Evidence for heme-heme excitonic coupling in the Soret circular dichroism of hemoglobin. AB - In order to study interdimer heme-heme electronic interactions in human hemoglobin, the Soret circular dichroism spectrum of the carboxy adduct is measured as a function of protein concentration, the spectrum at the highest concentration representing primarily that of alpha2beta2 tetramers (93%) and the lowest concentration representing primarily alphabeta dimers (68%). The tetramer dimer difference spectrum, obtained using singular value decomposition and linear least squares fitting from a matrix of CD spectra measured at ten concentrations, is roughly conservative, with a larger negative lobe at shorter wavelengths and a peak-to-trough magnitude that is 18% of the tetramer's maximum Soret CD magnitude. It is tentatively assigned to heme-heme excitonic interactions on the basis of theoretical predictions by R. W. Woody [(1985) in Optical Properties and Structure of Tetrapyrroles (Blauer, G., and Sund, H., Eds.), pp. 239-256, Walter de Gruyter, New York]. PMID- 9207207 TI - Photochemical formation of singlet molecular oxygen ((1)O2) in illuminated 6 methylcoumarin solutions. AB - Use of the fragrance 6-methylcoumarin (6-MC) in cosmetic products has declined significantly due to numerous reports of photoallergic contact dermatitis associated with its use. We have determined that 6-MC undergoes direct photolysis with an estimated half-life of 83 minutes when illuminated with mid-latitude U.S., noon-centered, equinox sunlight and a quantum yield for photolysis at 313 nm of phi = 3 x 10(-3). The work presented here also provides evidence that singlet molecular oxygen ((1)O2) is formed in illuminated solutions containing 6 MC. An estimated value of phi = 0.01 is reported for the (1)O2 quantum yield at 313 nm. Formation of (1)O2 is significant because it is known to react with a variety of biomolecules and it is possible that (1)O2 formation is at least partially responsible for reports of 6-MC photoallergenicity and phototoxicity. PMID- 9207208 TI - Citrulline-argininosuccinate-arginine cycle coupled to Ca2+-signaling in rat pancreatic beta-cells. AB - Pancreatic beta-cells possess nitric oxide (NO) synthases (NOSs) which synthesize NO and L-citrulline from L-arginine. The present study was designed to explore the mechanism of citrulline and arginine metabolism in beta-cells and its possible coupling to beta-cell functions. The enzymes involved in citrulline arginine metabolism, argininosuccinate synthetase (ASS), argininosuccinate lyase (ASL), and NOS were expressed in rat islets and insulinoma HIT T15 cells. In the presence of stimulatory glucose, L-citrulline and L-argininosuccinate at physiological concentrations (0.1-1 mM) increased cytosolic Ca2+ concentration ([Ca2+]i) in rat beta-cells. The citrulline-induced [Ca2+]i increase was inhibited by a NOS inhibitor, N(G)-monomethyl-L-arginine (NMMA). L-citrulline also stimulated NO production in HIT cells, which was inhibited by NMMA. In conclusion, L-citrulline is metabolized by ASS-ASL-NOS cycle to produce NO, which in turn increases [Ca2+]i in beta-cells. PMID- 9207209 TI - An alternative form of nucleolysin binds to a T-cluster DNA in the silencer element of platelet factor 4 gene. AB - The cDNA of a T-cluster binding protein (TCBP) has been cloned using the Southwestern method. The cDNA sequence of TCBP reveals that it has 78% homology to that of nucleolysin, a factor involved in apoptosis in cytolytic lymphocyte target cells. In particular, the 0.8kb sequences of the 5'-half of both cDNAs were identical. However, nucleolysin has a lysosome-targeting motif at the carboxy terminus, while TCBP has a hydrophobic sequence instead. Southern blot experiments have revealed that both cDNA sequences existed in the same YAC clone derived from chromosome 10. This strongly suggests that the TCBP cDNA is an alternatively spliced product of the nucleolysin/TCBP gene. The TCBP mRNA is ubiquitously expressed, not only in megakaryocytic cells but also in other hematopoietic cells. However, when HEL cells were induced to differentiate to megakaryocytes by DMSO, TCBP mRNA was reduced, while PF4 gene expression increased simultaneously. Gel mobility shift analysis demonstrated that recombinant TCBP specifically bound to the T-cluster and the proximal T-rich region of the PF4 promoter. Co-transfection experiments showed that TCBP reduced the gene expression from the PF4 promoter. On the other hand, TCBP did not affect expression from the PF4 promoter in which the T-cluster and the T-rich region had been removed. These results indicate that TCBP may participate in the regulation of PF4 gene expression by binding to the T-cluster and the T-rich sequence. PMID- 9207210 TI - Reconstitution and characterization of a divergent plastocyanin from the photosynthetic prokaryote, Prochlorothrix hollandica, expressed in Escherichia coli. AB - Plastocyanin (PC) is a copper protein that serves as a mobile electron carrier between cytochrome f and Photosystem I in the light reactions of photosynthesis. Despite large variability in amino acid sequences and isoelectric points, PCs from cyanobacterial and chloroplast sources reveal considerable similarities with respect to their secondary and tertiary structures. In this paper, we have expressed in Escherichia coli a PC from the prokaryote Prochlorothrix hollandica, and efficiently reconstituted the protein with copper under conditions yielding the characteristics of a native holoPC, as judged by redox titration (Eo' = +376 mV), near and far UV circular dichroism, and electron paramagnetic resonance (EPR) spectroscopy. By comparison of amino acid sequences, P. hollandica PC is the most divergent homolog identified to date, and analysis of this reconstituted preparation may reveal new insights as to the structural requirements for electron transport between the PC copper center and neighboring reaction partners. PMID- 9207211 TI - Pancreatic GAPDH gene expression during ontogeny and acute pancreatitis induced by caerulein. AB - Recent studies indicated that expression of the housekeeping gene GAPDH is highly regulated during proliferation and differentiation. The objective of this study was to characterize by Northern blot the GAPDH mRNA expression in rat pancreas development and regeneration following acute pancreatitis induction by caerulein. Pancreatic GAPDH mRNA levels were the highest between fetal day 19 and the 11 postnatal day; they decreased to their lowest level after weaning on day 26. In acute pancreatitis, GAPDH mRNA levels were clearly increased 18 h after its initiation, were maximal during the first two days of induction and then decreased to control values after 9 days. These data demonstrate that overexpression of GAPDH may be implicated in pancreatic development, maturation and pancreas regeneration after acute pancreatitis. PMID- 9207212 TI - Specific inhibitor for prolyl endopeptidase suppresses the generation of amyloid beta protein in NG108-15 cells. AB - A potent and specific prolyl endopeptidase inhibitor, JTP-4819, was used to investigate the role of prolyl endopeptidase in the generation of amyloid beta protein (A beta) from APP by NG108-15 cells. Synthetic substrates, 7-(succinyl Ile-Ala)-4-methylcoumarinamide and Z(Val-Lys-Met)-4-methylcoumarinamide, respectively, corresponding to the C-terminal and N-terminal portions of A beta, were cleaved by NG108-15 cell lysates. Cleavage of the C-terminal portion, but not the N-terminal, was inhibited by JTP-4819 (IC50 = 0.6 nM). Western blot analysis showed that the A beta level in the culture medium of NG108-15 cells was increased by serum deprivation. JTP-4819 caused concentration (>10(-9) M)- and time-dependent inhibition of this serum deprivation-induced increase of A beta without having any effect on the level of the secretory form of APP. Using both specific anti-A beta (1-40) and anti-A beta (1-42) antisera, the A beta that increased with serum deprivation was confirmed to be A beta (1-40), suggesting that it might be produced by conversion of A beta (1-42) to A beta (1-40). These findings indicate that prolyl endopeptidase may be a key enzyme in the production of A beta by NG108-15 cells and that A beta secretion can be modulated by a prolyl endopeptidase inhibitor. PMID- 9207213 TI - Regulation of osteoclastic bone resorption by glucose. AB - Osteoclasts degrade bone by pumping molar quantities of HCl to dissolve the calcium salts of bone, an energy intensive process evidently supported by abundant mitochondria. This is the first study to directly examine the ability of various metabolites to serve as potential energy sources for osteoclastic bone resorption. Glucose, and to a lesser extent lactate, supported osteoclastic bone degradation. However, fatty acids (palmitate, myristate and stearate), essential amino acids plus 20 mM alanine, or ketone bodies (acetoacetate, beta hydroxybutyrate and alpha-ketoglutarate) did not support bone degradation. Resorption declined to 10-30% of glucose controls when fatty acids or ketoacids were substituted for glucose. Resorption was glucose concentration dependent, with maximal activity at approximately 7 mM (K(M) approximately 3 mM). Glucose transport was linear for approximately 15 minutes, specific for D-glucose, and inhibited by cytochalasin B. Osteoclasts cultured on bone transported glucose at almost twice the rate of those off bone (Vmax 23 versus 13 nmols/mg/min, respectively) and medium acid accumulation paralleled glucose uptake, while the K(M) was unchanged. We conclude that glucose is the principal energy source required for bone degradation. Further, characteristics of glucose transport are consistent with the hypothesis that fluctuations in serum glucose concentration are an important component in regulation of osteoclastic bone degradation. PMID- 9207214 TI - Cytoplasmic processing of human profilaggrin by active mu-calpain. AB - The differentiation of keratinocytes involves numerous steps including the formation of the cornified envelope and the aggregation of keratin filaments by filaggrin monomer molecules. In this study, we investigated whether mu-calpain is involved in the processing of profilaggrin to filaggrin monomers by using both an active mu-calpain specific antibody and a 27-mer synthetic calpastatin peptide, a cell-permeable calpain-specific inhibitor. Upon incubation of cultured keratinocytes with Ca2+ for 96 hours, active mu-calpain with a molecular mass of 76 kDa appeared preceded by an increase in mu-calpain mRNA. In synchrony with the appearance of active mu-calpain, the processing of profilaggrin occurred. Furthermore, the Ca2+-induced activation of mu-calpain and the processing of profilaggrin were affected by the addition of the synthetic calpastatin inhibitor. These results indicate that the activation of mu-calpain plays a major role in the profilaggrin processing. PMID- 9207215 TI - Roles of inhibitors of myosin light chain kinase and tyrosine kinase on cation influx in agonist-stimulated endothelial cells. AB - Agonist-stimulated Ca2+ influx is critically important to mediate the function of endothelial cells. It has been suggested that release of Ca2+ from internal stores activates Ca2+ influx across the plasma membrane. In the present study, we investigated the effects of ML-9, a myosin light-chain kinase (MLCK) inhibitor, and genistein, a tyrosine kinase inhibitor, on the agonist stimulated Ca2+ response in porcine aortic endothelial cells loaded with a Ca2+-sensitive dye, fura-2. ML-9 almost completely abolished Ca2+ influx, whereas genistein only partially attenuated Ca2+ entry. Both of them did not affect the mobilization of Ca2+ from internal stores. In contrast, genistein was more potent in the inhibition of Mn2+ influx than ML-9. These findings indicate the different selectivity for Ca2+ and Mn2+ in the cation entry pathway in agonist-stimulated endothelial cells. PMID- 9207216 TI - Oligoadenosine tracts favor nucleosome formation. AB - We have measured the ability of oligoadenosine tracts 25 base pairs in length to influence nucleosome formation. Such tracts can cause DNA to bind in nucleosomes at higher temperatures with a free energy up to 1 kcal/mol more favorable than heterogenous-sequence DNA. Furthermore, the position of the oligoadenosine tract affects the free energy of binding, with the most favorable position occurring at the dyad axis. PMID- 9207218 TI - Beta-helical fibrils from a model peptide. AB - A synthetic peptide, KLEG13 (Ac-KLKLKLELELELG-NH2), composed of alternating bulky hydrophilic and hydrophobic amino acid residues formed clear, viscous dispersions of fibrils in saline solutions. The fibrils had a uniform diameter of 2 nm as measured on electron micrographs of negatively stained preparations. 13C solid state nuclear magnetic resonance spectroscopy of the fibrils indicated the presence of a beta-conformation. Circular dichroic spectra of the dispersion of fibrils were essentially identical to the calculated spectrum of a 100% beta helix. Space-filling CPK models of a proposed beta-helical conformation of the peptide, in which the leucine side chains form a hydrophobic core and the hydrophilic lysine and glutamate side chains extend outwards from the helix, had a diameter consistent with the observed 2-nm diameter of the fibrils. This study may have implications regarding the structure of amyloid fibrils. PMID- 9207217 TI - Evaluation of clinical and environmental anti-estrogens with human estrogen receptor expressed in Saccharomyces cerevisiae: a novel role for ABC-cassette transporters in mediating anti-estrogenic activity. AB - The effectiveness of anti-estrogens in treating estrogen-dependent diseases is limited by the acquired resistance of some diseases to anti-estrogens. This effect could occur by the export of anti-estrogens by cell membrane transport proteins. To study this phenomenon we have expressed human estrogen receptor (hER) and an estrogen-sensitive reporter in wild-type yeast and two transport defective strains. In the wild-type strain, the most effective anti-estrogen was nafoxidine. 4-Hydroxy tamoxifen and clomiphene were inactive whereas tamoxifen had significant inhibitory activity in the wild-type strain. Using a strain missing the ABC-cassette transporter Snq2, clomiphene had anti-estrogenic activity. 4-Hydroxy tamoxifen had anti-estrogenic activity only in yeast lacking the transporter Pdr5. Whole cell binding assays indicated that 4-hydroxy tamoxifen is exported by Pdr5. Environmental chemicals such as polychlorinated biphenyls function as partial estrogens and anti-estrogens in yeast. In the absence of Pdr5 or Snq2, the estrogenic activity of 4-hydroxy, 2',4',6'-trichloro biphenyl (3-PCB) was substantially reduced in comparison to its activity in the wild-type strain. Interestingly, the antiestrogenic activity of 3-PCB was equivalent in the wild-type and transporter-defective strains. Our results suggest a novel role for ABC-cassette transporters in regulating the activity of clinical and environmental anti-estrogens. PMID- 9207219 TI - Fluoroaluminate induces pertussis toxin-sensitive protein phosphorylation: differences in MC3T3-E1 osteoblastic and NIH3T3 fibroblastic cells. AB - Fluoride is an acknowledged bone-forming agent that may act through stimulation of osteoblast proliferation. Fluoride's action on osteoblasts and bone is potentiated by aluminum, which can form a complex with fluoride (fluoroaluminate) and activate heterotrimeric G proteins. Here we examined signaling pathways activated by fluoroaluminate in MC3T3-E1 osteoblastic and in NIH3T3 fibroblastic cells. In MC3T3-E1 cells, fluoroaluminate induced a decrease in cAMP levels and an increase in MAP and p70 S6 kinase phosphorylations. These responses were partially or completely prevented by pertussis toxin, an inhibitor of G alpha i proteins. In NIH3T3 cells, fluoroaluminate induced weaker tyrosine and MAP kinase phosphorylations. Fluoroaluminate, but not PDGF, induced a long-lasting tyrosine phosphorylation of a 130 kDa protein only in MC3T3-E1 cells. The expression of G alpha i2, but not of G alpha s and G alpha q/11 proteins was about 10-fold higher in MC3T3-E1 cells. Thus, different signaling in osteoblastic and fibroblastic cells may be due to differential expression of G alpha i proteins and tyrosine kinase substrates and could underlie fluoride's pharmacological action in bone. PMID- 9207220 TI - Oxidation of caffeine to theobromine and theophylline is catalyzed primarily by flavin-containing monooxygenase in liver microsomes. AB - Upon N-demethylation of caffeine (CA) by rat and human liver microsomes, theobromine (TB), paraxanthine (PX), and theophylline (TP) are produced. The optimal pHs for the formation of TB, PX, and TP from CA by rat liver microsomes are 7.4 (most), 8.2 (minor) and 8.6 (moderate). At pH 7.4, PX is the primary metabolite formed and makes up 48% of the CA metabolites generated. In the presence of SKF525A, an inhibitor of P450 (CYP), the rates of TB, PX and TP production are inhibited by 32%, 68% and 42%, respectively. Alternatively, in the presence of methimazole, an inhibitor of flavin-containing monooxygenase (FMO), the rates of TB, PX and TP production are inhibited by 66%, 48% and 73%, respectively. In the presence of both SKF525A and methimazole, they are inhibited by 95%, 84% and 94%, respectively. With human liver microsomes, the CA is metabolized faster but is inhibited more extensively either by SKF525A (PX production) or by methimazole (TB production). Alternatively, when CA is metabolized at pH 8.6, the optimal pH of FMO catalyzed reaction, the rates of TB and TP formation are increased but the rate of PX production is decreased. Furthermore, at pH 8.6 and in the presence of methimazole, the rates of TB and TP formation are decreased by 82% and 95%, respectively. These results indicate that the FMO is responsible primarily for productions of TB and TP and the CYP for PX. PMID- 9207221 TI - p53 is associated with the nuclear envelope in mouse testis. AB - p53 has been postulated to play a role in meiosis as well as in the regulation of germ cell numbers by apoptosis. This study investigated the subcellular localization of p53 in the testis, including conditions known to induce germ cell apoptosis. Western blot analysis showed that p53 was enriched in the nuclear envelope fraction, and confocal microscopy confirmed that p53 was associated with the nuclear envelope of germ cells. Exposure of the testis to heat stress induced translocation of p53 into the nucleus. Nuclear envelope binding provides an optimal site for rapid entry of p53 into the nucleus, where it may act as a DNA binding protein to induce apoptosis or cell cycle arrest in response to appropriate stimuli. The nuclear envelope sequestration of p53 also provides a framework to understand how mitosis and meiosis in the testis may proceed despite high intracellular concentration of p53. PMID- 9207222 TI - Kinetics and metabolism of chloral hydrate in children: identification of dichloroacetate as a metabolite. AB - Chloral hydrate was introduced into therapeutics more than 100 years ago, and since then a number of kinetic and metabolic studies have been conducted on this drug. Trichloroethanol, its glucuronide and trichloroacetic acid have been identified as the metabolites of chloral hydrate. We now report the identification of dichloroacetate as a major product of chloral hydrate metabolism in children, in addition to trichloroethanol and trichloroacetic acid. Furthermore, pretreatment of children with chloral hydrate appears to retard the plasma clearance of dichloroacetate. PMID- 9207223 TI - CD137, a member of the tumor necrosis factor receptor family, is located on chromosome 1p36, in a cluster of related genes, and colocalizes with several malignancies. AB - CD137 (ILA/4-1BB) is a member of the tumor-necrosis-factor receptor family. Members of this receptor family and their structurally related ligands are important regulators of a wide variety of physiological processes and play an especially important role in the regulation of immune responses. CD137 regulates cell proliferation and survival of T-lymphocytes. Using Southern blot analysis and polymerase chain reaction, we localized the CD137 gene to chromosome 1p36. This chromosomal region harbors the genes of several other members of this receptor family and is associated with deletions and rearrangements in several malignancies. PMID- 9207224 TI - Association of protein phosphatase-1delta with the retinoblastoma protein and reversible phosphatase activation in mitotic HeLa cells and in cells released from mitosis. AB - The retinoblastoma gene product (pRb) is dephosphorylated at the exit from mitosis and protein phosphatase-1 (PP1) seems to be responsible for such dephosphorylation. Three isoforms of PP1 exist in mammalian cells, alpha, gamma1 and delta, with differential subcellular localization and potentially different targeting subunits and functions. In order to identify which isoform dephosphorylates pRb, we used isoform-specific antibodies and analyzed the association of the PP1 isoforms with pRb in nocodazole-blocked (mitotic) HeLa cells and in cells released from the mitotic block (early G1). PP1delta was found associated with the pRb immunoprecipitated from a mitotic cell extract, whereas neither PP1gamma1 nor PP1alpha were detected. In G1 cells progressively less pRb and of lower Mr was detected in anti-PP1delta immunocomplexes, and pRb had almost disappeared by 8 h. The PP1 associated with pRb was inactive at mitosis, but underwent a quick activation as cells exited from mitosis, with a peak at 1 h. Then the activity decreased progressively and disappeared by 8 h. [32P]labeled pRb, obtained from G2 cells, was dephosphorylated "in vitro" by PP1delta obtained from early G1 cells. Altogether, the results indicated that PP1delta associated with pRb and may be responsible for the phosphatase activity detected in the pRb complexes, supporting the hypothesis that PP1delta may be the isoform that dephosphorylates pRb. PMID- 9207225 TI - The protein tyrosine phosphatase LAR has a major impact on insulin receptor dephosphorylation. AB - Transmembrane protein tyrosine phosphatases (PTPases) may act as regulators of the insulin receptor. Supporting this hypothesis, antisense suppression of the PTPase LAR in McA-RH7777 hepatoma cells increased insulin receptor signaling (Kulas et. al., J. Biol. Chem. (1996) 271, 748-754). The effects of decreased LAR expression may be mediated by decreased dephosphorylation of the insulin receptor. The rate of insulin receptor dephosphorylation was examined in situ, following elution of surface bound insulin at pH 4.0. In LAR antisense cells, dephosphorylation was prolonged by 2.6-fold with a t(1/2) of 87+/-11 sec compared to a t(1/2) of 34+/-6 sec in control cells. EGF receptor dephosphorylation was also prolonged in LAR antisense cells. These results are further evidence that LAR is a physiological regulator of the insulin receptor and is consistent with its direct interaction with the tyrosine phosphorylated insulin receptor. PMID- 9207226 TI - Vascular endothelial growth factor increases endothelin-converting enzyme expression in vascular endothelial cells. AB - Endothelin-converting enzyme-1 (ECE-1) is a key enzyme in endothelin processing. Although it has been revealed that ECE-1 expression increases in vascular wall after balloon injury in vivo experimental models, the regulation of ECE-1 expression in vitro remains undefined. In this study, we demonstrated that the endothelial cell-specific mitogen, vascular endothelial growth factor (VEGF) increased ECE-1 expression in cultured bovine aortic endothelial cells (BAEC). Northern blot analysis revealed that VEGF increased ECE-1 mRNA expression in 12 24 hours by 2-fold in BAEC, and this effect was dose-dependent. VEGF also increased ECE-1 protein expression detected by immunoblotting in 36 hours in BAEC. Therefore, VEGF increased ECE-1 expression in BAEC, which suggests that ECE 1 induction by VEGF may be involved in endothelin-system upregulation under pathological conditions such as neointimal formation and atherosclerosis. PMID- 9207227 TI - Lack of nucleotide-promoted second messenger signaling responses in 1321N1 cells expressing the proposed P2Y receptor, p2y7. AB - A recently cloned G protein-coupled receptor (named the p2y7 receptor) with relatively low sequence identity to previously cloned P2Y receptors was proposed to be a member of this family of receptors on the basis of both a radioligand binding assay with [35S]dATP alphaS and an inositol phosphate response to ATP in COS-7 cells transiently transfected with receptor cDNA. Previous work in our laboratory has shown that [35S]dATP alphaS is not a general radioligand for the identification of P2Y receptors and that COS-7 cells express an endogenous P2Y receptor (P2Y2) that complicates the analysis of nucleotide-promoted inositol phosphate responses. Thus, data supporting inclusion of the p2y7 receptor in the P2Y family of receptors are equivocal. To determine unambiguously whether the p2y7 receptor is a P2Y receptor subtype, a p2y7 receptor bearing an epitope-tag at its NH2-terminus was expressed in 1321N1 cells and cell surface expression of the receptor was demonstrated by an intact cell-based ELISA. Cells shown to express epitope-tagged p2y7 receptors by ELISA were examined for their second messenger signaling properties in response to a range of nucleotides. ATP, UTP, ADP, UDP, and dATP alphaS had no effect on phospholipase C or adenylyl cyclase activities in cells expressing the p2y7 receptor. Experimental controls utilizing expression of other G protein-coupled receptors showed that 1321N1 cells displayed robust responses for each of these signaling pathways. These data, together with the low sequence identity of the p2y7 receptor to other P2Y receptors, indicate that the p2y7 is not a member of the P2Y family of signaling molecules. PMID- 9207228 TI - Isolation and immunoaffinity purification of biologically active plant natriuretic peptide. AB - It has recently been demonstrated that antibodies against atrial natriuretic peptides (ANP) recognise analogues in plants and that rat ANP binds specifically to isolated plant membranes and promotes stomatal guard cell opening in a concentration dependent manner. Here we report the isolation and immunoaffinity purification of plant natriuretic peptide (PNP) from ivy (Hedera helix) with rabbit anti-alpha-ANP (1-28) (human, canine) antiserum. We also demonstrate that immunoaffinity purified plant peptide induces stomatal opening in a concentration dependent manner. This is therefore the first report of an active indigenous peptide hormone in plants. We propose that PNPs are part of a signalling system that has evolved early in evolution and is involved in the regulation of ion transport and transpiration in plants. PMID- 9207229 TI - The enhancing effect of anionic alpha-helical peptide on cationic peptide mediating transfection systems. AB - A peptide consisting of 12 amino acids including 3 glutamic acids (LAEL-LAEL LAEL; 4(3)E) underwent pH-dependent conformational change from random coil to alpha-helix when the pH was decreased from 7.4 to 5.0 in the presence of egg PC. This alpha-helical 4(3)E had higher membrane-perturbation activity at acidic conditions compared with neutral conditions. When 4(3)E was incorporated with plasmid DNA-cationic peptide complex that utilizes an endocytosis pathway for uptake into cultured cells, high transfection efficiency was observed, indicating that 4(3)E can enhance the transfection activity of cationic peptide. It is likely that 4(3)E in the multi-complex of the plasmid DNA and the cationic peptide effectively disrupts the endosomal membrane and increases the population of the complex which could transfer to cytosol. The small lysosome-disruptive peptide is very probably useful as the enhancer molecule for the gene transfer techniques mediated by the endocytosis pathway. PMID- 9207230 TI - A methylation-dependent DNA-binding activity recognising the methylated promoter region of the mouse Xist gene. AB - Differential methylation of CpG sites in the promoter region of the mouse Xist gene is correlated with Xist expression and X-chromosome inactivation in the female. Using oligonucleotides encompassing the differentially methylated sites as probes in band-shift assays, we have identified a nuclear protein which binds to a specific region of the promoter (between base pairs -45 and -30 upstream from the transcription start site) only when CpG sites within the CG rich region (GCGCCGCGG, -44 to -36) are methylated. Competition experiments with methylated or unmethylated heterologous oligonucleotides demonstrate that the activity is sequence-specific as well as methylation-dependent. Analysis by Southwestern blot identifies a protein of approximately 100 kDa molecular weight and confirms strong binding to the methylated Xist promoter oligonucleotide. Using a 233bp Xist-promoter luciferase construct in which the cytosines in the three CpG sites in the -44 to -36 region are mutated to thymine, we have established that this region is required for transcription from the mouse Xist promoter. Therefore, we suggest that the binding of the 100kDa protein to the methylated sequence leads to repression of transcription from the methylated Xist allele, thus suggesting a role in the regulation of both imprinted and random Xist transcription and X chromosome inactivation. PMID- 9207231 TI - Effect of oncogene transformation of rat embryo cells on cellular oxygen consumption and glycolysis. AB - We found an unique effect of oncogene transfections on rat embryo cell (REF) respiration, glycolysis and radiation response. Radioresistance, defined as an increase in Do, increases for REF cells transfected with v-myc or H-ras oncogenes. The combination of both oncogenes confers the maximal radioresistance. Our work shows inhibition of oxygen uptake when cells are transfected with v-myc or H-ras alone. However, oxygen uptake increases when cells are transfected simultaneously with v-myc + H-ras (3.7,2.1,2.8). A higher oxygen consumption results from increased utilization of pyruvate via the Kreb's cycle. Succinate stimulates cellular oxygen consumption. The maximum stimulation of oxygen consumption by succinate occurred with v-myc + H-ras transfected cells. The glycolysis of the transfected cells is also altered by the oncogenes. Our glycolytic measurements indicate the H-ras oncogene causes the largest stimulation of glycolysis. Our data shows that transfection with oncogenes has a major effect on cellular glycolysis, oxidative metabolism as well as the subsequent radiation response. PMID- 9207232 TI - Scatter factor/hepatocyte growth factor expression enhances human glioblastoma tumorigenicity and growth. AB - We have shown previously that the multifunctional cytokine scatter factor/hepatocyte growth factor (SF/HGF) is elevated in human malignant gliomas. In this study we investigated how human SF/HGF expression affects the malignancy of the U373 human glioblastoma cell line in vivo and in vitro. Human SF/HGF gene transfer increased U373 glioblastoma tumorigenicity by > or = 20-fold and enhanced the growth rate of intracerebral U373 xenografts by 3- to 8-fold. SF/HGF expression had no effect on the proliferation of glioblastoma cell monolayers but increased their anchorage-independent colony formation in soft agar by 5- to 8 fold. These results are the first to show that SF/HGF expression by human glioblastoma cells enhances their growth dysregulation in vitro and malignancy in vivo. PMID- 9207233 TI - Correlation between loss of middle ear bones and altered goosecoid gene expression in the branchial region following retinoic acid treatment of mouse embryos in vivo. AB - The homeobox gene goosecoid marks the Spemann organizer in vertebrate gastrula embryos, and is expressed in the craniofacial region, body wall and limbs during organogenesis. Mouse mutants of goosecoid displayed a variety of phenotypes related to the expression pattern at mid-embryogenesis. These defects included loss of the tympanic ring and malformation of the malleus, phenotypes which were reminiscent of the teratogenic effects of retinoic acid (RA). Here we investigated the correlation of goosecoid gene expression and RA-teratogenicity following treatment of mouse embryos in vivo at embryonic day (E) 8 + 5 h. We found that goosecoid was specifically affected at E10.5 in branchial arches I and II. Expression was either reduced to background levels or restricted to the branchial cleft region. This change in goosecoid gene expression correlated with a loss of middle ear ossicles and a partial or complete deletion of the tympanic ring, suggesting a role for goosecoid in executing the RA teratogenic effects. PMID- 9207234 TI - ED2+ macrophages increase selectively myoblast proliferation in muscle cultures. AB - We have previously shown by coculturing myoblasts and macrophages that myotube formation is strongly increased in vitro by the presence of an acid stable, heat labile, soluble growth factor(s) secreted by macrophages. In this paper we obtained macrophages from peritoneal washing which also contained limited amounts of other cells such as lymphocytes and mesothelial cells. We here demonstrate that an ED2-positive (ED2+) macrophage subpopulation is responsible for myoblast enhanced proliferation. ED2+ macrophages were separated by a magnetic-activated cell sorter (MACS) using a monoclonal antibody against ED2, a membrane antigen peculiar to macrophages. Both ED2+ macrophages and their conditioned medium increased myotube formation when added to primary muscle cultures. Furthermore we demonstrate that muscle growth induced by macrophages is mainly the consequence of an increased myoblast proliferation by showing the presence of an increased number of MyoD-positive (MyoD+) myonuclei. PMID- 9207235 TI - Kupffer cells are a dominant site of uncoupling protein 2 expression in rat liver. AB - The mechanisms underlying thermogenesis in liver are not well understood. They may involve proteins related to the mitochondrial uncoupling protein (UCP1) of brown adipocytes. In this paper, it is demonstrated that UCP1 is not expressed in any liver cell type of rat while UCP2, a recently cloned homologue of UCP1, is expressed at a very high level in Kupffer cells but not in hepatocytes. This high level of expression of UCP2 in Kupffer cells allowed cross immunoreactivity with antibodies directed against UCP1. This cross reactivity was confirmed by the detection of UCP2 with anti-UCP1 antibody, in western blotting analysis of transfected yeasts expressing rat UCP2. The high level expression of UCP2 in Kupffer cells suggests a particular function of UCP2 in macrophages. PMID- 9207236 TI - Functional consequences of constitutively active alpha2A-adrenergic receptor expression in 3T3F442A preadipocytes and adipocytes. AB - The functional consequences of a constitutively active mutated (CAM) human alpha2C10-adrenergic receptor (AR) stably expressed in the 3T3F442A preadipose cell line were analysed at both preadipocyte and adipocyte stages. At the preadipocyte stage, CAMalpha2C10-AR reproduced (in the absence of agonist) and amplified (in the presence of agonist) most of the cellular responses promoted by agonist-stimulated wild type alpha2C10-AR (increased preadipocyte proliferation, tyrosyl-phosphorylation of the Mitogen Activated Protein Kinases, resistance to serum-deprivation-induced cell retraction, inhibition of differentiation). In contrast, at the adipocyte stage, CAMalpha2C10-AR expression did not reproduced nor amplified the alpha2-adrenergic-dependent antilipolysis, but conversely led to a down-regulation of alpha i subunits of the Gi proteins and to an increase in the maximal response to lipolytic agents. Our results indicate that long term activation of intracellular signals by CAM-receptors not only lead to the expected cellular responses normally generated by agonist-stimulated wild type receptors, but can also lead to unexpected responses resulting from long term compensatory adaptations. PMID- 9207237 TI - Interleukin-8 selectively enhances cytopathic effect (CPE) induced by positive strand RNA viruses in the human WISH cell line. AB - Interleukin-8 (IL-8), a proinflammatory chemokine, is induced by viruses and appears in circulation during viral infections. We found that IL-8 enhanced cytopathic effect induced by the positive strand RNA virus, encephalomyocarditis virus (EMCV), in the human WISH cell line. The enhancement was dependent on IL-8 dose and virus dose and was reversible by specific monoclonal antibodies to IL-8. The chemokine was also able to increase EMC viral RNA synthesis and infectious virus yield. This IL-8 enhancing action was not observed in the case of the negative strand RNA virus, vesicular stomatitis virus (VSV), in WISH cells. We examined the activity of constitutive 2',5'-oligoadenylate synthetase (OAS), a pathway that was implicated in protection from EMCV but not VSV. The IL-8 action in EMCV-infected cells, unlike VSV-infected cells, was associated with decreased OAS activity in a manner that was independent of OAS gene expression. Understanding mechanisms of cytokine enhancement of viral activity may lead to novel ways to control viral infections. PMID- 9207238 TI - Identification of alternative first exons of NADH-cytochrome b5 reductase gene expressed ubiquitously in human cells. AB - Two forms of NADH-cytochrome b5 reductase (b5R), soluble and membrane-bound, are known. A hypothesis that the human soluble form b5R is generated through post translational processing of the membrane-bound form was previously proposed. In this study, the 5'-rapid amplification of cDNA ends for human reticulocyte, liver, brain, and HL-60 cell mRNAs revealed the ubiquitous presence of an alternative type of human b5R mRNA which can probably be translated into the soluble form b5R directly; however, the erythroid-specific transcript of the b5R gene was not found. This type of b5R mRNA initiating from at least two sites contains a non-coding new first exon located between the first two exons of the human b5R gene identified before. In addition, this new first exon shares 62% homology with the first exon and its 3'-flanking intron sequences of rat erythroid-specific b5R mRNA, whereas the 5'-flanking region of the new first exon possesses features of house-keeping gene. These results might be important to understand the regulation mechanism of human b5R biosynthesis and divergent evolution of the gene regulation. PMID- 9207239 TI - L-type Ca2+ channel-insulin-like growth factor-1 receptor signaling impairment in aging rat skeletal muscle. AB - The present study investigates the modulation of skeletal muscle L-type Ca2+ channel receptor in response to insulin-like growth factor-1 receptor (IGF-1R) activation. Single extensor digitorum longus and multifiber preparations were isolated from 7- (young), 14- (middle-age) and 28-(old) month- Fisher 344 X Brown Norway rats. Calcium current was potentiated in fibers from young and middle-age rats due to a -13 mV shift in half-activation potential. Fibers from old animals failed to show current potentiation in response to IGF-1R activation. IGF-1 induced a ten-fold increase in the phosphorylation of the L-type Ca2+ channel alpha1 subunit in young and middle-age fibers but failed to induce phosphorylation in old fibers. Addition of 0.5 mM Ca2+ increased the IGF-1 induced phosphorylation in young and middle-age fibers three fold but not in old fibers. The tyrosine kinase inhibitor, genistein, and the PKC inhibitor peptide, 19-36, decreased IGF-1 induced phosphorylation of alpha1 subunit to 15% in young and middle-age fibers but failed to inhibit phosphorylation in old fibers. These results demonstrate that the IGF-1-L-type Ca2+ channel alpha1 subunit signaling is impaired in skeletal muscle fibers from old animals due to alterations in the trk-PKC pathway. PMID- 9207240 TI - Effect of triiodothyronine on muscle cell differentiation and blood glucose level in hyperglycemic KK mice. AB - KK mice are genetically diabetic animals, showing glucose intolerance and insulin resistance. We examined the effects of 3,3',5-triiodo-L-thyronine (T3) on the blood glucose level and on mRNA levels of muscle cell differentiation markers in hyperglycemic KK mice. T3 treatment (T1, 1 mg; T3, 3 mg; T10, 10 mg/kg/day) of KK mice for 4 days caused a decrease in blood glucose level by 11%, 25%, and 24%, respectively, without affecting body weight. Skeletal muscle of mice treated with T3 (T10) showed a 98% increase in the mRNA level of the glucose transporter isotype 4 (Glut4). In contrast, T3 treatment did not affect the mRNA level of the isotype 1 (Glut1) transporter. The mRNA level of a muscle cell specific differentiation marker, MyoD, showed a significant increase in the T3 treatment group with an accompanying enhancement of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA level. These results suggest that T3 stimulates muscle cell differentiation in vivo, concomitant with a stimulation of cellular glucose metabolism, thus decreasing the blood glucose level in hyperglycemic KK mice. PMID- 9207241 TI - Cloning of a SH3 domain-containing proline-rich protein, p85SPR, and its localization in focal adhesion. AB - A mouse thymus cDNA expression library was screened with monoclonal antibody (mAb), B16-5 which binds to common epitope in SH3 domains of phospholipase C gamma 1 (PLC-gamma 1) and Nck. We have determined the complete nucleotide sequence of one of several positive clones. The 4,172 bp cDNA clone (GenBank Accession No. U96634) encodes a SH3 domain-containing protein of 646 amino acids. Besides the SH3 domain, the predicted protein has a proline-rich region, nuclear localization signals, and leucine zipper motifs. The expressed protein in Sf9 insect cell exhibits a polypeptide of 85 kDa on SDS-PAGE. The protein is widely distributed in rat tissue with an especially high level of expression in brain and testis. Interestingly, the specific antibodies detected four related proteins of different size (75, 85, 90 and 105 kDa) in brain. In A431 cell, p85SPR is enriched at focal adhesion points indicating that the protein may interact with protein(s) in focal complexes. PMID- 9207242 TI - Structural modifications of RNA influence the 5' exoribonucleolytic hydrolysis by XRN1 and HKE1 of Saccharomyces cerevisiae. AB - Two 5' exoribonucleases, XRN1 and HKE1, of Saccharomyces cerevisiae have been found to have very important cellular roles, XRN1 playing a key role in mRNA turnover and HKE1 in pre-rRNA processing. Here, an analysis of strong secondary structures in RNA that cause blocks or stalls (accumulation of RNA fragments that are shortened from the 5' end to the site of the secondary structure insertion) in the processive exoribonucleolytic hydrolysis reactions is reported. With both enzymes, oligo(G) tracts of lengths 18, 16, and 9 stall quite effectively, and the stalls are close to the start of the oligo(G) stretch. Two strong stem-loop structures cause measurable but low-level stalls with both enzymes. If the stem loop structure is placed close to the 5' end of the RNA, substantial inhibition of overall RNA hydrolysis occurs with HKE1 and less, but measurable, inhibition with XRN1. RNA structural modification caused by protein complexing has been investigated by using poly(A) binding protein. The hydrolysis of poly(A) by XRN1 is inhibited by poly(A) binding protein, while HKE1 activity is not affected. PMID- 9207243 TI - A selection system to study protein-RNA interactions: functional display of HIV-1 Tat protein on filamentous bacteriophage M13. AB - The transactivator protein (Tat) of the human immunodeficiency virus (HIV) is a key regulatory protein in the viral replication cycle and belongs to the RNA binding proteins of the arginine-rich motif (ARM) family. Very little is known about their mechanism of RNA recognition. To study the principles of RNA-protein recognition we constructed a system to display HIV-1 Tat on the surface of the filamentous bacteriophage M13. HIV-1 Tat (1-72) and a mutant Tat lacking five cysteine residues were cloned into the pAK phagemid system, which allows fusion of the tat gene to a supershort version of the gene for minor M13 coat protein. Expression of the resulting fusion proteins was shown via western blot analysis. Phages displaying functional Tat could be selected from phages without Tat or with a non-functional Tat variant via binding to biotinylated TAR using streptavidin coated paramagnetic beads. By randomizing certain amino acid positions of Tat and screening of the resulting phage libraries for affinity and specificity, we are now able to study the role and importance of amino acids of HIV-1 Tat for affinity and specificity to TAR RNA. PMID- 9207244 TI - Cooperation between the Fc epsilonR1 and formyl peptide receptor signaling pathways in RBL(FPR) cells: the contribution of receptor-specific Ca2+ mobilization responses. AB - RBL(FPR) mast cells express the tyrosine kinase-coupled IgE receptor, Fc epsilonR1, and the G-protein-coupled formyl peptide receptor, FPR. Fc epsilonR1 crosslinking causes Ca2+ stores release, Ca2+ influx, Ins(1,4,5)P3 production and secretion. FPR ligation also mobilizes Ca2+, but without measurable Ins(1,4,5)P3 production or secretion. Co-stimulating the FPR and Fc epsilonR1 induces more Ins(1,4,5)P3 production and secretion than Fc epsilonR1 cross-linking alone. Costimulation also produces more rapid and sustained Ca2+ responses than are generated by Fc epsilonR1 activation alone. We identified multiple differences between the FPR- and Fc epsilonR1-coupled Ca2+ responses, including a more rapid Ca2+ spike response to FPR ligation; intracellular Ca2+ stores that are empty following Fc epsilonR1 crosslinking but partially full following FPR activation; a more sustained Ca2+ influx response to Fc epsilonR1 crosslinking; and the immediate inhibition of stimulated Ca2+ influx by FPR antagonists but not by monovalent ligand that terminates Fc epsilonR1 crosslinking. We hypothesize that the interaction of receptor-specific Ca2+ mobilization pathways contributes to the FPR-mediated potentiation of Fc epsilonR1-coupled secretion. PMID- 9207245 TI - mRNA expression of HNF-4 isoforms and of HNF-1alpha/HNF-1beta variants and differentiation of human cell lines that mimic highly specialized phenotypes of intestinal epithelium. AB - The mRNA expression of HNF-4 isoforms and the ratio of HNF-1alpha/HNF-1beta variants in cell lines representing highly specialized phenotypes of human intestinal epithelium were studied by RT-PCR. A strong rise in expression of HNF 4 isoforms alpha2, alpha4 and gamma correlates with commitment into highly differentiated enterocyte-like phenotype of Caco-2 cells which best mimic enterocytes, whereas only isoform alpha4 expression is high in the less differentiated HT-29 G- cells. These increased expressions are not encountered in the highly differentiated mucous-secreting HT-29 MTX cells. Differentiation into highly specialized enterocyte-like Caco-2 cells and mucous-secreting HT-29 MTX cells is accompanied by a moderate rise in HNF-1 without change in the ratio of its variants. Our data corroborate those of Spath et al. (Mol. Cell. Biol., 1997, 17, 1913) in hepatoma cells and suggest that HNF-4 isoforms alpha2, alpha4 and gamma play a major role in the differentiation of enterocytes. PMID- 9207246 TI - The unique cytoplasmic domain of the human integrin variant beta4E is produced by partial retention of intronic sequences. AB - A novel cytoplasmic splice variant of the human beta4 integrin subunit has been identified by reverse transcription polymerase chain reaction using mRNA from cultured keratinocytes as the template. This fifth beta4 variant, called beta4E, is expressed in a wide variety of tissues including the epidermis, lung, duodenum, heart, spleen and stomach and in several human epithelial cell lines. The beta4E cDNA contains an insert of 37 base pairs which produces a frame shift in the sequence encoding the beta4 cytoplasmic domain and generates a new stop codon after a stretch of cDNA encoding a unique 114-amino acid peptide. Analysis of the genomic organization at the site of this insertion in the human beta4 gene reveals that beta4E is produced by partial retention of an intron in the final transcript. PMID- 9207247 TI - Intranuclear translocation of phospholipase C beta2 during HL-60 myeloid differentiation. AB - Phospholipases C (PLC) beta3, gamma1, and gamma2 were detected in nuclei of HL-60 promyelocitic leukaemia cells. When HL-60 cells undergo terminal myeloid differentiation in the presence of ATRA, the beta2 isoform appeared inside nuclei and was up-regulated until 72 hours of ATRA treatment. The beta3 isozyme was also increased until 72 hours and both isoforms lowered their intranuclear amount at 96 hours and following days of treatment. By contrast PLC gamma1 and gamma2 progressively increased in the nucleus during granulocytic differentiation even after 72 hours of treatment. Terminal differentiation was characterised by the expression of high levels of PLC gamma1 and gamma2 and by low levels of PLC beta2 and beta3 in the nucleus. PIP2 and PIP hydrolysis paralleled the prevalence of the beta or gamma subfamily, respectively. Moreover, at all the examined times no changes of PLCs in the whole cell were detectable, indicating a de novo nuclear translocation of the beta2 and an increased accumulation of beta3, gamma1, and gamma2 isoforms. Thus, the intranuclear presence, expression, and activity of PLC isozymes, which are modulated during differentiation of HL-60 cells, implicate a role for nuclear phosphoinositide signalling in the process of cell maturation. In particular the nuclear translocation of PLC beta2 candidates this PLC as a key enzyme in the granulocytic differentiative commitment of HL-60 cells. PMID- 9207248 TI - The regulation of pHi in osteoclasts is dependent on the culture substrate and on the stage of the resorption cycle. AB - The present study was performed to clarify the role of vacuolar H+-ATPase in the regulation of the intracellular pH (pHi) in osteoclasts. Bafilomycin A1 or amiloride were added to rat osteoclast cultures to block the H+-ATPases and Na+/H+-exchangers, respectively. Addition of 10(-8) M bafilomycin A1 to osteoclasts cultured on bone induced a rapid decrease of the pHi, while addition of amiloride had only a minor effect. The response to bafilomycin A1 appeared simultaneously with resorption activity and was abolished when resorption was inhibited by calcitonin. Osteoclasts on bone recovered from acid loading caused by propionate in the presence of amiloride, while bafilomycin A1 inhibited this recovery almost completely. The pHi of osteoclasts cultured on glass responded to the addition of amiloride, but was not effected by even high concentrations of bafilomycin A1. In contrast, as little as 10(-10) M bafilomycin A1 caused accumulation of large vesicles in the cytoplasm. PMID- 9207249 TI - Processing of Alzheimer's amyloid precursor protein during H2O2-induced apoptosis in human neuronal cells. AB - The processing of Alzheimer's amyloid precursor protein was studied by Western blotting during H2O2 induced apoptosis in cultures of human neuroblastoma cells. A new 5.5 kDa fragment putatively containing intact A beta was detected and found to be highly associated with apoptosis. The results suggest a possible vicious cycle involving H2O2, A beta and apoptosis which may contribute to the neuronal death mechanism in Alzheimer's Disease. PMID- 9207250 TI - Function of P-glycoprotein expressed in placenta and mole. AB - We examined the expression of P-glycoprotein in human placentas and hydatidiform moles. Trophoblasts in all the examined placentas and moles expressed P glycoprotein, and the size of the P-glycoprotein was smaller than that in multidrug-resistant human epidermoid carcinoma KB cells. The P-glycoprotein in the placenta and mole was photolabeled with [3H]azidopine, and [3H]vincristine was transported in an ATP-dependent manner into membrane vesicles prepared from trophoblasts that expressed P-glycoprotein. These findings indicate that P glycoprotein expressed in trophoblasts has a drug binding site(s) and the ability to transport vincristine, suggesting that P-glycoprotein in the placenta protects the fetus from xenobiotics and confers drug resistance on moles. PMID- 9207252 TI - Should DCC be deleted as tumor-suppressor candidate? PMID- 9207251 TI - RalA interacts directly with the Arf-responsive, PIP2-dependent phospholipase D1. AB - RalA GTPase associates with a phospholipase D (PLD) that is activated in v-Src- and v-Ras-transformed cells. Two mammalian PLDs were recently cloned: PLD1, which is activated by Arf family GTPases and dependent upon phosphatidylinositol-4,5 bisphosphate (PIP2), and PLD2, which is also dependent upon PIP2, but not stimulated by Arf. Another PLD has been described that is stimulated by oleate. Evidence is provided that the RalA-assiciated PLD is PLD1. First, the PLD precipitated by RalA from murine fibroblasts was stimulated by Arf, dependent upon PIP2, and inhibited by oleate. Second, immobilized RalA precipitated PLD1 from sf9 insect cells overexpressing PLD1. Third, a series of RalA mutants precipitated PLD activity from both PLD1-expressing insect cells and murine fibroblasts with the same efficiency. And finally, immobilized RalA precipitated PLD1 from a purified PLD1 preparation. These data argue that RalA associates directly with the Arf-responsive, PIP2-dependent PLD1. PMID- 9207253 TI - A new congenital diarrhea gene. PMID- 9207254 TI - Genetic testing: the time is now. PMID- 9207255 TI - Image of the month. Intrahepatic biliary cystadenocarcinoma with hepatic metastases and peritoneal dissemination. PMID- 9207256 TI - Health-related quality of life after ileoanal pull-through evaluation and assessment of new health status measures. AB - BACKGROUND & AIMS: Health-related quality of life (HRQL) after proctocolectomy is a critical parameter for management decisions in patients with chronic pancolitis. The aim of this study was to evaluate the HRQL of patients with ileoanal pull-through and to validate new, easy-to-administer HRQL measures. METHODS: The Sickness Impact Profile (SIP), Short Form 36 (SF-36), Rating Form of Inflammatory Bowel Disease (IBD) Patient Concerns (RFIPC), and the time trade-off (TTO) were used to measure HRQL of pull-through patients. The SF-36 and the RFIPC were validated. RESULTS: HRQL of patients with ileoanal pull-through was better than that of a national sample of patients with IBD (SIP and RFIPC) and similar to that of a normal population (SF-36). Physical and psychosocial subscales of the SF-36 correlated with the SIP, affirming the construct validity of the SF-36. The RFIPC results correlated with the SIP and SF-36 results, suggesting that it is also a valid health status measure for these patients. TTO results correlated with the physical subscales of the SIP and SF-36, reflecting the impact of physical health on this group. CONCLUSIONS: HRQL of patients with ileoanal pull through is excellent. The SF-36 and RFIPC are valid health status measures that can be used by clinicians and researchers in these patients. PMID- 9207257 TI - Helicobacter pylori infection and chronic gastric acid hyposecretion. AB - BACKGROUND & AIMS: We have identified a subgroup of Helicobacter pylori-infected subjects with low or absent gastric acid output. The aim of this study was to document the morphological and functional abnormalities in these subjects and to assess the effect of eradicating the infection. METHODS: The 16 hypochlorhydric subjects (6 men) had a mean age of 55 years (range, 36-79 years). They underwent a 14C-urea breath test, H. pylori serology, fasting gastrin, gastric autoantibodies, gastroscopy with antral and body biopsies, and measurement of peak acid output to pentagastrin (PAO(PG)). Their histology was compared with that of age- and sex-matched duodenal ulcer and nonulcer dyspepsia patients (16 each). H. pylori infection was eradicated in the hypochlorhydric subjects, and the investigations were repeated 6 months later. RESULTS: Compared with controls, the hypochlorhydric subjects had less dense H. pylori colonization, body predominant colonization and gastritis, and increased prevalence of body atrophy and intestinal metaplasia. Median PAO(PG) before eradication in the hypochlorhydric subjects was 1.1 mmol/h and increased to 12.6 mmol/h after eradication (P < 0.001), with no significant change in body atrophy or intestinal metaplasia. CONCLUSIONS: In some subjects, chronic H. pylori infection produces a body-predominant gastritis and profound suppression of gastric acid secretion that is partially reversible with eradication therapy. PMID- 9207258 TI - Effect of Helicobacter pylori eradication on gastritis in relation to cagA: a prospective 1-year follow-up study. AB - BACKGROUND & AIMS: Whether Helicobacter pylori eradication resolves intestinal metaplasia and atrophy and whether infection with cagA+ H. pylori is related to a specific clinical outcome are not known. The aim of this study was to investigate the role of H. pylori eradication on the course of intestinal metaplasia (IM) and atrophy in relation to cagA. METHODS: In a large prospective study, the cagA status of H. pylori isolated from consecutive dyspeptic patients was related to clinical outcome before and 1 year after successful eradication of H. pylori. At pretreatment and 4-6 weeks and on average 1 year after eradication therapy, the degree of gastritis and the status of H. pylori were assessed by culture and histopathology. RESULTS: Specimens of cagA+ H. pylori were recovered from 122 of 155 (79%) patients infected with H. pylori. Pretreatment degrees of gastritis activity, superficial epithelial damage, IM, and atrophy were significantly greater in patients infected with cagA+ H. pylori (P < 0.001). After successful eradication of H. pylori, a significant improvement of activity of gastritis and superficial epithelial damage occurred (P < 0.001), but the degree of IM and atrophy did not change, irrespective of the cagA status. CONCLUSIONS: The usefulness of H. pylori eradication to revert precancerous lesions such as IM and atrophy after 1-year follow-up is questionable. PMID- 9207259 TI - Serum 13C-bicarbonate assay for the diagnosis of gastric Helicobacter pylori infection and response to treatment. AB - BACKGROUND & AIMS: Current serological tests for Helicobacter pylori (HP) infection are not useful in assessing active infection or eradication. The feasibility, sensitivity, and specificity of serum 13C-bicarbonate (13C-HCO3) measurement in determining gastric HP before and after eradication by antibiotics were investigated. METHODS: Twenty-seven symptomatic patients underwent endoscopy, biopsy, and CLOtest. Patients then consumed a 13C-urea-rich meal; serum was collected before and 1 hour after meal ingestion for quantitative determination of 13C by mass spectrometry. Postprandial fractional elevation of 13C (delta 13C-HCO3) was correlated with endoscopy, histology, and CLOtest at baseline and at 4 and 8 weeks after therapy. RESULTS: delta 13C-HCO3 +/- SEM was 17.02 +/- 2.94 in HP-positive patients and 2.77 +/- 044 in HP-negative patients (P < 0.0001). In HP-positive patients who responded to therapy, the mean change was initially 20.5 +/- 3.1, 3.2 +/- 0.9 1 month after therapy, and 2.8 +/- 0.4 2 months after therapy (P < 0.001). The overall sensitivity of this method was 90.6% (95% confidence interval, 74.9-98.0), and its specificity was 85.7% (95% confidence interval, 42.1%-99.6%). delta 13C-HCO3 correlated positively with the degree of histological gastritis and the number of HP organisms. CONCLUSIONS: Serum 13C-HCO3 analysis is a novel, simple, and noninvasive method for diagnosis and assessment of eradication of HP infection. PMID- 9207260 TI - Intragastric distribution and gastric emptying assessed by three-dimensional ultrasonography. AB - BACKGROUND & AIMS: Three-dimensional (3D) ultrasound imaging of the total stomach volume has not yet been achieved. The aim of this study was to investigate whether a magnetic position sensor system for acquisition of 3D ultrasonograms could be used to determine gastric emptying rates and intragastric distribution. METHODS: A system for position and orientation measurement was interfaced to an ultrasound scanner. In vitro accuracy was evaluated by scanning a porcine stomach. Fourteen volunteers, with a median age of 35 years, were scanned fasting and postcibally by two-dimensional (2D) and 3D ultrasound after ingesting a 500 mL soup meal. RESULTS: This 3D system yielded a strong correlation (r = 0.997) between true and estimated volumes in vitro. The limits of agreement were 9.1:70.1 mL in the volume range 1200-1900 mL. The intersubject variability of the total gastric volumes ranged from 12.5% to 46.0%, less than for antral area variability. The average half-emptying time was 22.1 +/- 3.8 minutes. Intragastric distribution of the meal, expressed as proximal distal volume, varied on average from 3.6 +/- 2.1 (5 minutes postpradially) to 2.7 +/- 1.9 (30 minutes postprandially). CONCLUSIONS: This 3D ultrasound system using magnetic scanhead tracking showed excellent in vitro accuracy, calculated gastric emptying rates more precisely than by 2D ultrasound, and enabled estimation of intragastric distribution of a soup meal. PMID- 9207261 TI - Identification of human brain loci processing esophageal sensation using positron emission tomography. AB - BACKGROUND & AIMS: Brain loci that process human esophageal sensation remain unidentified. The aim of this study was to identify the brain loci that process nonpainful and painful human esophageal sensation. METHODS: In 8 healthy subjects (7 men; age range, 24-47 years), distal esophageal stimulation was performed by repeatedly inflating a balloon at volumes that produced either no sensation, definite sensation, or pain. Two positron emission tomography scans were performed for each sensation using H2(15)O. Magnetic resonance brain scans were also performed in each subject, and the positron emission tomography data were coregistered with magnetic resonance scans. Analysis of covariance-corrected t images showing the contrasts definite sensation-baseline, pain-baseline, and pain definite sensation were created. RESULTS: Nonpainful stimulation elicited bilateral activations along the central sulcus, insular cortex, and frontal/parietal operculum (P < 0.01). Painful stimulation produced more intense activations of the same areas and additional activation of the right anterior insular cortex and the anterior cingulate gyrus. Multiple areas of decreased activation were also observed; prominent among these was the right prefrontal cortex, which was inhibited during both nonpainful and painful stimulation. CONCLUSIONS: Esophageal sensation activates bilaterally the insula, primary somatosensory cortex, and operculum. The right anterior insular cortex and anterior cingulate gyrus process esophageal pain. PMID- 9207262 TI - Physiological hyperglycemia slows gastric emptying in normal subjects and patients with insulin-dependent diabetes mellitus. AB - BACKGROUND & AIMS: Marked hyperglycemia slows and hypoglycemia accelerates gastric emptying. The aim of this study was to determine the effect of physiological changes in blood glucose gastric emptying. METHODS: In 8 healthy subjects and 9 patients with insulin-dependent diabetes mellitus (IDDM) without gastrointestinal tract symptoms or evidence of neuropathy, gastric emptying of a mixed meal was measured by scintigraphy. Using an insulin-glucose clamp, the blood glucose concentration was stabilized at 4 and 8 mmol/L on 2 separate days. RESULTS: The intragastric retention of the solid meal component at 100 minutes was 55.2% +/- 4.5% at 8 mmol/L vs. 36.7% +/- 5.5% at 4 mmol/L (P = 0.004) in normal subjects and 44.2% +/- 4.2% vs. 35.7% +/- 4.2% (P = 0.004) in patients with IDDM. The time taken for 50% emptying of the liquid meal was 57.0 +/- 10.8 minutes at 8 mmol/L vs. 32.2 +/- 12.6 minutes at 4 mmol/L (P = 0.002) in normal subjects and 41.3 +/- 3.4 minutes vs. 29.1 +/- 3.5 minutes (P = 0.002) in patients with IDDM. CONCLUSIONS: Changes in blood glucose within the normal postprandial range have a significant impact on gastric emptying in both normal subjects and patients with IDDM. PMID- 9207264 TI - Absence of effect of Lewis A and Lewis B expression on adherence of Helicobacter pylori to human gastric cells. AB - BACKGROUND & AIMS: Lewis b blood group antigen and antibodies to Lewis b inhibit the binding of stationary-phase Helicobacter pylori organisms to fixed sections of gastric tissue. The aim of this study was to determine the effect of expression of Lewis a and Lewis b on binding of H. pylori to primary gastric cells. METHODS: ABO and Lewis blood types were determined for 13 individuals. Cells were isolated from gastric biopsy specimens by collagenase digestion. Lewis antigen expression and adherence of H. pylori to the cells were quantitated using flow cytometry. RESULTS: Two of the three nonsecretors were found to express Lewis b on their cells. Nineteen of 19 individuals expressed Lewis a on their cells and 18 of 19 expressed Lewis b. The percentage of cells expressing Lewis antigens varied from individual to individual. H. pylori binding was independent of expression of Lewis antigens. Preincubation of cells with antibodies to Lewis antigens did not inhibit the adherence. CONCLUSIONS: H. pylori adheres to isolated human gastric cells in a manner that is not dependent on Lewis antigen expression on these cells, and expression of Lewis antigens on the surface of gastric cells is not dependent on Lewis antigen expression on erythrocytes. PMID- 9207263 TI - Propeptide of type I procollagen is predictive of posttreatment bone mass gain in adult celiac disease. AB - BACKGROUND & AIMS: Adult celiac disease is associated with osteopenia, which is not always reversible after gluten-free diet (GFD). A prospective study was conducted to evaluate whether pretreatment indices of bone and mineral metabolism are predictive of the extent of bone mass gain after diet. METHODS: Lumbar and femoral bone mineral density (z-score) and serum levels of parathyroid hormone, 1,25-dihydroxycholecalciferol, COOH-terminal propeptide of type I procollagen (PICP), and COOH-terminal telopeptide of type I collagen (ICTP) were measured in 20 celiac patients at diagnosis and after 2 years of GFD. RESULTS: All patients showed a posttreatment improvement in bone mass and in serum levels of indices of bone and mineral metabolism. Nevertheless, only in 12 of 20 patients was this improvement at least equal to half the SD of the z-score, which equals a gain of at least 5% in bone mass. Pretreatment levels of PICP strictly correlated with the increase in lumbar (r(s) = 0.92; P < 0.001) and femoral z-scores (r(s) = 0.89; P < 0.001). Only in patients with basal PICP above the normal range did the z-score increase after GFD by at least half the SD. CONCLUSIONS: In adult celiac disease, a high rate of osteosynthetic activity before treatment is predictive of the satisfactory recovery of bone mass after GFD. PMID- 9207265 TI - Interferon gamma and interleukin 1, but not interferon alfa, inhibit rotavirus entry into human intestinal cell lines. AB - BACKGROUND & AIMS: Rotavirus, an important agent of gastroenteritis in children, causes diarrhea by infecting differentiated villus enterocytes in the small intestine. The aim of this study was to determine whether cytokines that can be expressed by mucosal cells have an effect on the rotavirus susceptibility of cultured human enterocytes. METHODS: Caco-2 and HT-29 cells were pretreated with various cytokines before challenge with rotavirus. RESULTS: Interleukin (IL)-1, interferon (IFN)-alpha, and IFN-gamma pretreatment led to a dose-dependent resistance to rotavirus infection. Maximum effects occurred after 72 hours of pretreatment, whereas no detectable inhibition occurred with <12 hours of pretreatment. Liposomal transfection of single-shelled and double-shelled rotavirus particles bypassed the block to rotavirus replication in IFN-gamma- and IL-1-treated but not IFN-alpha-treated cells. Binding studies with purified, metabolically labeled rotavirus showed no significant difference among IFN-gamma- and IFN-alpha-treated and control Caco-2 cells. Viral entry into Caco-2 cells was significantly inhibited by IFN-gamma and IL-1 but not IFN-alpha. CONCLUSIONS: IFN alpha and IFN-gamma induce rotavirus resistance by different mechanisms, suggesting that cytokines play a role in host defense against viral agents by changing the phenotype of intestinal epithelial cells. PMID- 9207266 TI - Electrophysiological properties of human carcinoid cells of the gut. AB - BACKGROUND & AIMS: Because of their diffuse distribution, neuroendocrine cells of the gut have not been isolated successfully for electrophysiological characterization. We therefore established primary cell cultures from surgically resected human carcinoids and investigated them electrophysiologically. METHODS: The neuroendocrine identity of the isolated gut tumor cells was determined immunocytochemically. The electrophysiological properties of the cells were studied by the patch-clamp technique. RESULTS: The primary cell cultures expressed neurofilament proteins, cytokeratins, and key proteins of the secretion machinery. Spontaneous action potentials were observed in most cells. Using the whole-cell mode of the patch-clamp technique, tetrodotoxin-sensitive voltage gated sodium currents as well as voltage-gated calcium currents were identified. Calcium channel currents were carried mainly by dihydropyridine-sensitive, L-type calcium channels. The L-type calcium channel currents were also partially blocked by the omega-conotoxins GVIA and MVIIC. Moreover, omega-agatoxin IVA reversibly reduced a component of the calcium channel currents, indicating that neuroendocrine gut tumor cells express different types of voltage-gated calcium channels. In addition, somatostatin was found to inhibit partially the voltage dependent calcium channel currents and thus calcium-dependent hormone release. CONCLUSIONS: Carcinoid cells of the human gut are electrically excitable cells. They express voltage-dependent sodium and calcium channels as well as somatostatin receptors. PMID- 9207267 TI - Goblet cell autoantibodies in patients with inflammatory bowel disease and their first-degree relatives. AB - BACKGROUND & AIMS: Crohn's disease and ulcerative colitis show a familial aggregation. In both diseases, anti-goblet cell autoantibodies (GABs) have been described. The aim of this study was to define the role of GABs in the pathogenesis of inflammatory bowel disease. METHODS: The study population comprised 61 patients with ulcerative colitis, 76 patients with Crohn's disease, 101 first-degree relatives of patients with ulcerative colitis, and 105 first degree relatives of patients with Crohn's disease. Thirty-five patients with infectious enterocolitis and 56 healthy unrelated subjects served as controls. Autoantibodies were detected by indirect immunofluorescence. RESULTS: Thirty-nine percent of patients with ulcerative colitis (24 of 61) and 30% of patients with Crohn's disease (23 of 76) were positive for GABs. GABs were detected in 21% (21 of 101) of first-degree relatives of patients with ulcerative colitis and in 19% (20 of 105) of first-degree relatives of patients with Crohn's disease. In patients with infectious enterocolitis and in healthy controls, GABs were seen in 3% (1 of 35) and 2% (1 of 56), respectively. The differences between control groups and both groups of patients or their first-degree relatives were significant. CONCLUSIONS: The high prevalence in first-degree relatives suggests that GABs may represent a marker characterizing susceptibility to inflammatory bowel disease. PMID- 9207268 TI - Elevated serum levels and reduced immunohistochemical expression of thrombomodulin in active ulcerative colitis. AB - BACKGROUND & AIMS: The pathogenesis of ulcerative colitis and Crohn's disease is still unclear. Vascular injury has been suggested as a potential pathogenetic mechanism. Serum thrombomodulin is a marker of endothelial cell injury. The aim of this study was to determine the relevance of increased serum thrombomodulin levels for assessing disease activity in inflammatory bowel disease. As a potential cause of serum thrombomodulin level increase, the loss of local vascular thrombomodulin expression was investigated immunohistochemically. METHODS: Thrombomodulin levels were determined by enzyme-linked immunosorbent assay in sera from patients with ulcerative colitis, Crohn's disease, Schistosoma mansoni infection, and infectious diarrhea and controls. The vascular expression of thrombomodulin was investigated immunohistochemically in fresh frozen transmural specimens of normal, Crohn's, and ulcerative colitis bowel samples. RESULTS: Significantly elevated serum thrombomodulin levels were only detected in active ulcerative colitis and infectious diarrhea complicated by septicemia. A marked and general loss of vascular endothelial cell thrombomodulin expression was found immunohistochemically in inflamed bowel tissues. Graded by a newly established thrombomodulin staining index, this was significantly more marked in ulcerative colitis than Crohn's disease. CONCLUSIONS: Serum thrombomodulin proved to be a novel marker of disease activity in ulcerative colitis closely related to local vascular endothelial cell damage, which might be a relevant pathophysiological feature of ulcerative colitis. PMID- 9207270 TI - Genetic alterations in sporadic and Crohn's-associated adenocarcinomas of the small intestine. AB - BACKGROUND & AIMS: Small intestinal carcinomas are rare but occur with increased incidence in Crohn's disease. The aim of this study was to elucidate the genetic alterations. METHODS: Mutations and deletions involved in colorectal carcinoma were studied in sporadic and Crohn's-associated intestinal carcinomas and precursors. RESULTS: c-K-ras mutations were present in all four sporadic carcinomas with contiguous adenomas, in only 18% without adenomas (P = 0.01), in 43% of Crohn's-associated carcinomas, and in 14% of dysplasias. Overexpression of p53 gene product and/or 17p allelic loss were present in 47% of sporadic carcinomas and 33% of contiguous adenomas and in 71% of Crohn's-associated carcinomas and 43% of dysplasias. In contrast, allelic losses of 5q (adenomatous polyposis coli [APC] gene region) and 18q (deleted in colorectal cancer [DCC] gene region) were rare. DNA replication errors (RERs) were present in 13% of sporadic carcinomas and in the carcinoma and dysplasias of 1 patient with Crohn's disease (14%), but mutations in the transforming growth factor beta type II receptor (TGFbeta RII) gene were absent. CONCLUSIONS: Accumulation of ras and p53 alterations occurs during the adenoma/dysplasia-carcinoma sequence in small intestinal carcinogenesis, but a ras-independent pathway may also exist. The infrequent loss of the APC and DCC regions and the absence of TGFbeta RII gene mutation in RER-positive neoplasms contrast with colorectal carcinogenesis. PMID- 9207269 TI - Distinct cytokine patterns in early and chronic ileal lesions of Crohn's disease. AB - BACKGROUND & AIMS: Chronic intestinal lesions of patients with Crohn's disease (CD) are associated with a T helper (Th) 1-type cytokine profile, including high levels of interleukin 2 (IL-2) and interferon gamma (IFN-gamma). However, the mechanisms involved in the pathogenesis of the early mucosal lesions are poorly known. The aim of this study was to examine the pattern of Th1- and Th2-type (IL 4, IL-5, and IL-13) cytokines in the early ileal lesions occurring in patients with CD 3 months after ileal resection and ileocolonic anastomosis. Cytokines were also examined in the chronic ileal lesions to look for cytokine patterns related to disease progression. METHODS: Ileal biopsy specimens were obtained from 17 patients with CD and 11 controls. Mucosal IL-2, IFN-gamma, IL-4, IL-5, and IL-13 messenger RNA (mRNA) was evaluated by competitive reverse-transcription polymerase chain reaction. RESULTS: The early ileal lesions of patients with CD were associated with a significant increase of IL-4 mRNA and a decrease of IFN gamma mRNA compared with the normal mucosa of patients with CD or controls. A Th1 type pattern was observed in the chronic ileal lesions. CONCLUSIONS: Divergent cytokine patterns are observed during different clinical stages of CD. These observations need to be considered in the development of newer specific therapeutic agents to prevent CD recurrences. PMID- 9207271 TI - Extracellular matrix composition and gene expression in collagenous colitis. AB - BACKGROUND & AIMS: Collagenous colitis is a rare diarrheal disease of unknown pathophysiology that is histologically defined by subepithelial bandlike structures. The objective of this study was to elucidate the biochemical composition and the origin of the bandlike structures in collagenous colitis. METHODS: Immunohistochemical and in situ hybridization analyses were performed on endoscopic specimens using specific antibodies and riboprobes for collagen types I, III, IV, and VI and for the glycoprotein tenascin. RESULTS: In collagenous colitis, the mucosal matrix with the exception of the bands retained a normal architecture and extracellular matrix composition. The bands stained most prominently for type VI collagen and tenascin. Less abundant staining for both proteins was also found in the subepithelial matrix of the normal mucosa. In situ hybridization showed no significant increase in collagen type VI messenger RNA expression in cells around and entrapped in the bands in collagenous colitis compared with normal specimens. CONCLUSIONS: The results support the suggestion that collagenous colitis is a localized alteration of the extracellular matrix, which involves the pericryptal-subepithelial myofibroblast sheath. The data suggest that reduced matrix degradation and not overactivation of matrix synthesis may be the reason for the subepithelial accumulation of matrix proteins. PMID- 9207272 TI - Mesenchymal cells stimulate human intestinal intraepithelial lymphocytes. AB - BACKGROUND & AIMS: Intraepithelial lymphocytes (IELs) from human intestinal mucosa proliferate minimally to T-cell stimuli. Optimal growth may depend on factors that are missing in vitro, such as accessory cells. The aim of this study was to determine whether mesenchymal cells costimulate IELs. METHODS: IELs were isolated from human jejunum and cultured with fibroblasts or smooth muscle cells (mesenchymal cell models for mucosal myofibroblasts) and various T-cell stimuli. Proliferation was determined by [3H]thymidine incorporation, and interleukin 2 (IL-2) production was measured by enzyme-linked immunosorbent assay. Surface molecules were detected by immunofluorescence and flow cytometry. RESULTS: The proliferative responses of IELs to mitogen (phytohemagglutinin), superantigen (staphylococcal enterotoxin B), or anti-CD3 antibody were increased greatly by coculture with mesenchymal cells, while only slightly by peripheral-blood monocytes, the classical antigen-presenting cells. IL-2 production and receptor expression also increased. Mesenchymal cell costimulation of IEL growth required direct contact between the two cell types and was partly dependent on the integrin alpha4beta1 (very late activation 4[VLA-4]) and major histocompatibility complex (MHC) class I, as their respective antibodies blocked the effect. The surface molecules B7 (CD80), CD2, and MHC class II were not involved. CONCLUSIONS: Optimal IEL growth depends on their contact with mesenchymal cells, an interaction that is mediated by VLA-4 and MHC class I. In mucosal immunity, basement membrane myofibroblasts likely serve this role. PMID- 9207273 TI - Interleukin 10 prevents cytokine-induced disruption of T84 monolayer barrier integrity and limits chloride secretion. AB - BACKGROUND & AIMS: The proinflammatory cytokine interferon gamma (IFN-gamma) disrupts epithelial barrier integrity and attenuates secretagogue-induced chloride secretion. This study tested the efficacy of the anti-inflammatory cytokine interleukin 10 (IL-10) in maintaining epithelial barrier and chloride secretory function in the presence of IFN-gamma. METHODS: T84 epithelial cell monolayers were treated with IL-10, IFN-gamma, or IFN-gamma plus IL-10. Monolayer barrier integrity was assessed by measurements of electrical conductance, unidirectional mannitol and inulin fluxes, and tight junctional charge selectivity in Ussing chambers. Short-circuit current (Isc) was measured in response to carbachol and forskolin stimulation. RESULTS: IL-10 attenuated the IFN-gamma-induced increase in electrical conductance and totally prevented the IFN-gamma-induced increase in mannitol and inulin fluxes. IL-10 did not prevent the IFN-gamma-induced abolishment of tight junctional charge selectivity but did attenuate the total increase in sodium and chloride permeability. IFN-gamma and IL-10 both separately reduced peak forskolin and carbachol-stimulated Isc. IL-10 pretreatment further enhanced the IFN-gamma-induced reduction in secretagogue induced Isc. CONCLUSIONS: In T84 epithelial monolayers, IL-10 maintains the size, but not the charge, selectivity of the epithelial tight junction in the presence of IFN-gamma. In addition, both IL-10 and IFN-gamma limit carbachol and forskolin induced increase in Isc. PMID- 9207274 TI - CD95 (APO-1/Fas)-mediated apoptosis in colon epithelial cells: a possible role in ulcerative colitis. AB - BACKGROUND & AIMS: Ligation of CD95 (APO-1/Fas) by antibody or CD95 ligand (CD95L) induces apoptosis and, in some cell lines, growth. Normal colonic epithelial cells constitutively express CD95. The function of CD95 on colonocytes is unknown. The aim of this study was to elucidate the role of epithelial CD95 in the normal colon and in ulcerative colitis. METHODS: Intact colonic crypts were isolated, and the effects of CD95 ligation in vitro were studied. CD95L expressing cells and apoptotic cells were detected in situ by RNA hybridization, immunohistochemistry, and DNA nick end labeling. RESULTS: On CD95 ligation, isolated colonic crypt cells underwent apoptosis within 4 hours. No growth promoting effect was observed. In normal colon, CD95L expression was restricted to few mononuclear cells randomly scattered within the lamina propria. Therefore, the CD95-CD95L system is very unlikely to operate in the regeneration of the colonic epithelium. However, in ulcerative colitis, the number of interstitial CD95L+ cells and the frequency of apoptosis in both lamina propria and epithelium were increased considerably. Further, a focal association of subepithelial CD95L+ mononuclear cells and epithelial apoptosis was observed. CONCLUSIONS: In ulcerative colitis, soluble CD95L-mediated epithelial apoptosis may lead to a breakdown of the epithelial barrier function facilitating the invasion of pathogenic microorganisms. PMID- 9207275 TI - Fas-mediated cytotoxicity by intestinal intraepithelial lymphocytes during acute graft-versus-host disease in mice. AB - BACKGROUND & AIMS: Host-derived intestinal intraepithelial lymphocytes (IELs) increase in number during acute graft-versus-host disease (GVHD) in mice. In the present study, we examined Fas-mediated cytotoxicity by host-derived IELs against Fas-expressing target cells to see whether Fas/Fas ligand (Fas-L) interaction is involved in the pathogenesis of enteropathy during acute GVHD. METHODS: Acute GVHD was induced by injection of parental spleen cells into nonirradiated F1 mice. The expression of Fas antigen on the intestinal epithelial cells (IECs) was examined by flow cytometry, and the expression of messenger RNA (mRNA) for Fas-L, interleukin 2, and interferon gamma in host-derived IELs was assessed by reverse transcription polymerase chain reaction. Fas-mediated cytotoxicity by host derived IELs was assessed using Fas-transfected cells, IECs, and Fas immunoglobulin Fc fusion protein (Fas Fc). RESULTS: Fas antigen was constitutively expressed on the cell surface of IECs before and after GVHD induction. Although Fas-L mRNA was not detected or detected scarcely in either alphabeta or gammadelta IELs before GVHD induction, both IELs expressed high levels of mRNA for Fas-L and interferon gamma after GVHD induction. Host-derived IELs during acute GVHD showed cytotoxicity against Fas-transfected target cells and IECs, which was partly blocked by addition of Fas Fc. CONCLUSIONS: Fas/Fas-L mediated cytotoxicity by host-derived IELs may be partly responsible for the enteropathy during acute GVHD. PMID- 9207276 TI - Bromelain prevents secretion caused by Vibrio cholerae and Escherichia coli enterotoxins in rabbit ileum in vitro. AB - BACKGROUND & AIMS: Diarrhea is a major cause of illness and death in children and young animals. The aim of this study was to investigate the possible therapeutic effect of bromelain, a proteolytic extract obtained from pineapple stems on bacterial toxin and second-messenger agonist-induced intestinal secretion. METHODS: The effect of bromelain pretreatment on short-circuit responses to Escherichia coli heat-labile enterotoxin, heat-stable enterotoxin, and Vibrio cholerae cholera toxin was evaluated in rabbit ileum mounted in Ussing chambers. RESULTS: Bromelain was 62% effective in preventing heat-labile enterotoxin induced secretion, 51% effective against cholera toxin, and 35% effective against heat-stable enterotoxin [corrected]. Bromelain also prevented secretory changes caused by prostaglandin E2, theophylline, calcium-ionophore A23187, 8 bromoadenosine 3':5'-cyclic monophosphate, and 8-bromoguanosine 3':5'-cyclic monophosphate, well-known intracellular mediators of ion secretion. The efficacy of bromelain was not caused by reduced tissue viability resulting from its proteolytic effects on enterocytes, indicated by experiments measuring uptakes of nutrients into intestinal cells and experiments measuring short-circuit responses to glucose. CONCLUSIONS: Bromelain prevents intestinal fluid secretion mediated by secretagogues that act via adenosine 3':5'-cyclic monophosphate, guanosine 3':5'-cyclic monophosphate, and calcium-dependent signaling cascades. It may be clinically useful as an antidiarrheal drug. PMID- 9207277 TI - Local immunoglobulin G antibodies in the stomach may contribute to immunity against Helicobacter infection in mice. AB - BACKGROUND & AIMS: Orogastric immunization of mice with Helicobacter antigens, together with a mucosal adjuvant (cholera toxin), has been shown to confer immunity in the Helicobacter felis infection model. The aim of the study was to investigate the humoral immune responses associated with immunity and to compare these with responses in H. felis-infected mice. METHODS: Enzyme-linked immunoassays were used to characterize the antibody-secreting cells and antibodies present at mucosal and systemic sites in mice. Animals were immunized orally with either whole-cell H. felis sonicates or Helicobacter pylon urease or heat-shock proteins. RESULTS: Infection of mice with H. felis preferentially induced the recruitment of plasma cells committed to immunoglobulin (Ig) A synthesis in salivary gland and gastric tissues. Antigen-specific IgA was the major antibody class detected in mucosal secretions recovered from these tissues. In contrast, immunization of mice against H. felis infection induced the proliferation of large numbers of IgG-secreting cells, as well as the synthesis of local IgG antibodies, in the gastric mucosa of the animals. Protection against H. felis infection occurred in the absence of gastric IgA responses in sonicate immunized mice. CONCLUSIONS: It is proposed that locally synthesized specific IgG antibodies contribute to immunity against gastric Helicobacter infection. PMID- 9207278 TI - Induction of cyclooxygenase 1 and 2 in the rat stomach during endotoxemia: role in resistance to damage. AB - BACKGROUND & AIMS: Prostaglandins and nitric oxide are key mediators of gastric mucosal defense. Endotoxemia alters gastric resistance to damage, but little is known of the effects of chronic endotoxemia on the expression of prostaglandin and nitric oxide synthases (NOSs). The effects of short- vs. long-term administration of endotoxin on gastric resistance to damage and on expression of NOS and prostaglandin synthesis were compared. METHODS: Rats were treated with short- or long-term bacterial endotoxin, after which susceptibility to ethanol induced damage was assessed. The effects of various inhibitors of prostaglandin and NOS were examined. Expression of gastric NOS and cyclooxygenase (COX) messenger RNA (mRNA) were examined. RESULTS: Repeated administration of endotoxin increased gastric resistance to ethanol- but not indomethacin-induced injury. Indomethacin, but not a highly selective COX-2 inhibitor or an inducible NOS inhibitor, abolished long-term endotoxin-induced gastric resistance to injury. Expression of mRNA for both COX-1 and -2, but not for endothelial or inducible NOS, were significantly increased after long-term endotoxin administration. CONCLUSIONS: Repeated exposure to endotoxin resulted in increased resistance of the gastric mucosa to injury through a prostaglandin-dependent pathway. These prostaglandins were produced via COX-1, which like COX-2, is induced by endotoxin administration. PMID- 9207279 TI - Inhibition of glucose absorption in the rat jejunum: a novel action of alpha-D glucosidase inhibitors. AB - BACKGROUND & AIMS: alpha-D-Glucosidase inhibitors act primarily by decreasing disaccharide hydrolysis and thus reduce the amount of free monosaccharides available for absorption. A novel action of alpha-D-glucosidase inhibitors is presented, indicating a direct effect on free glucose absorption by the rat jejunum. METHODS: The jejunum was isolated and free hexose was measured using in vivo single-pass luminal perfusion and dual vascular and luminal single-pass in vitro perfusion. Xenopus oocytes were injected with RNA transcript encoding recombinant sodium-glucose cotransporter 1, and uptake of 3H-labeled 3-O-methyl-D glucopyranose (3-O-MG) was assessed. RESULTS: Acarbose (0.1 mg/mL), added to the lumen, decreased D-glucose absorption by 20% in vivo. Addition of 0.1 or 1.0 mg/mL acarbose to the lumen in vitro decreased the appearance of 3-O-MG in the vascular effluent by 28% and 60%, respectively. Accumulation of D-glucose within the enterocytes was decreased significantly by 67% and 79% when acarbose (1 mg/mL) or phloridzin (2 mmol/L), respectively, were present in the luminal perfusate. In contrast, acarbose did not affect the transport rate of free D fructose and did not inhibit 3-O-MG uptake in oocytes expressing sodium-glucose cotransporter 1. CONCLUSIONS: The findings indicate that alpha-D-glucosidase inhibitors act specifically on the entry of free glucose into the enterocyte, an additional means by which they can reduce postprandial hyperglycemia. PMID- 9207280 TI - Wilson's disease in patients presenting with liver disease: a diagnostic challenge. AB - BACKGROUND & AIMS: In patients with Wilson's disease presenting with liver involvement, the correct diagnosis is often missed or delayed. The aim of this study was to find an algorithm for diagnosis of this difficult patient group. METHODS: Clinical and laboratory findings of 55 patients with Wilson's disease were evaluated at diagnosis before treatment. Presenting symptom was chronic liver disease in 17 patients, fulminant hepatic failure in 5 patients, hemolysis in 3 patients, and neurological disease in 20 patients, and 10 patients were detected by family screening (siblings). Evaluation included neurological and ophthalmologic examination, routine laboratory tests, and parameters of copper metabolism including liver copper content in 43 liver biopsy specimens. RESULTS: In the whole group, serum ceruloplasmin level was <20 mg/dL in 73%, urinary copper excretion was increased in 88%, and liver copper content was elevated in 91% at diagnosis. Kayser-Fleischer rings were detected in 55%. In contrast to patients with neurological disease (90% Kayser-Fleischer rings, 85% low ceruloplasmin), only 65% of patients presenting with liver disease were diagnosed by these typical findings. Ceruloplasmin levels were lower in patients with Kayser-Fleischer rings or with neurological disturbances than in patients without these symptoms. CONCLUSIONS: The commonly used clinical and laboratory parameters are not sufficient to exclude the diagnosis of Wilson's disease in patients with liver disease of unknown origin. PMID- 9207282 TI - Treatment of biliary colic with diclofenac: a randomized, double-blind, placebo controlled study. AB - BACKGROUND & AIMS: Nonsteroidal anti-inflammatory drugs (NSAIDs) have been used to relieve biliary colic. Follow-up was limited in previous studies, and the role of NSAIDs in the natural history of biliary colic has not been clarified. The purpose of this study was to evaluate the efficacy of diclofenac, a potent NSAID, in the the immediate symptomatic relief of biliary colic and the prevention of cholelithiasis-related complications. METHODS: Fifty-three patients with cholelithiasis and biliary colic were enrolled in this randomized, double-blind, placebo-controlled study. They received a single 75-mg (3 mL) intramuscular injection of diclofenac (n = 27) or similarly administered 3 mL of saline (n = 26). All patients were followed up for at least 3 days. The effect of either treatment was assessed by changes in the severity of pain and the development of cholelithiasis-related complications. RESULTS: Complete relief of pain was obtained in 21 diclofenac and in 7 placebo patients; progression to acute cholecystitis was observed in 4 and 11 patients, respectively. Fewer overall complications were observed in the diclofenac group. CONCLUSIONS: In patients with cholelithiasis who present with biliary colic, a single 75-mg intramuscular dose of diclofenac can provide satisfactory pain relief and decrease substantially the rate of progression to acute cholecystitis. PMID- 9207281 TI - Etidronate versus fluoride for treatment of osteopenia in primary biliary cirrhosis: preliminary results after 2 years. AB - BACKGROUND & AIMS: Because osteopenia increases morbidity of primary biliary cirrhosis (PBC), the effects of cyclical etidronate vs. sodium fluoride on bone mass were compared in patients with PBC. METHODS: Thirty-two women with PBC were randomly assigned to receive etidronate (400 mg/day during 14 days every 3 months) or fluoride (50 mg/day, enteric-coated tablets). Bone mineral density of the lumbar spine and proximal femur were measured initially and every 6 months. Bone fractures were also evaluated. RESULTS: Sixteen patients were allocated into each group, which were comparable with respect to the severity of PBC and osteopenia. Thirteen patients with etidronate and 10 patients with fluoride completed 2 years in the study. In the etidronate group, bone mineral density increased in the lumbar spine (P = 0.02) and did not change in the proximal femur. In the fluoride group, lumbar bone mineral density did not change but femoral bone mass decreased, particularly in the Ward's triangle. Two patients in the fluoride and none in the etidronate group developed new vertebral fractures, and the number of new nonvertebral fractures was similar in both groups. Neither treatment impaired liver function or cholestasis. CONCLUSIONS: Cyclical etidronate is more effective and better tolerated than sodium fluoride in preventing bone loss in PBC. PMID- 9207283 TI - Ethyl propionate is more effective and less cytotoxic than methyl tert-butyl ether for topical gallstone dissolution. AB - BACKGROUND & AIMS: Ethyl propionate and isopropyl acetate were identified as gallstone solvents with more favorable physicochemical properties than the currently used solvent methyl tert-butyl ether (MTBE). In this study, their efficacy and toxicity were compared. METHODS: To compare efficacy, matched stones from 33 patients were subjected to dissolution with each solvent. To evaluate cytotoxicity, jejunal segments of the anesthetized rat were exposed to each solvent or saline; the segments were then perfused with markers for active absorption and passive permeability. RESULTS: For 23 gallstone sets that dissolved completely with all three solvents, the average dissolution time was shorter with ethyl propionate (38 +/- 8 minutes) than with MTBE (60 +/- 13 minutes) (P = 0.03) or isopropyl acetate (55 +/- 12 minutes) (P < 0.001). Four stones did not dissolve with ethyl propionate, seven with MTBE, and eight with isopropyl acetate. After 2 minutes of exposure to the solvents, the dry weight of the segments decreased by 36% after MTBE but was unchanged after the other two solvents (P < 0.001). MTBE caused more inhibition of active absorption than the other solvents (P < 0.001) and a greater increase in passive permeation (P < 0.03). CONCLUSIONS: Ethyl propionate and isopropyl acetate are less toxic to the intestinal mucosa than MTBE, and ethyl propionate is more effective for gallstone dissolution. PMID- 9207284 TI - Hepatic hyperplasia and cancer in rats: metabolic alterations associated with cell growth. AB - BACKGROUND & AIMS: We showed previously that the peroxisome proliferators di(2 ethylhexyl)phthalate (DEHP), clofibrate, and 4-chloro-6-(2,3 xylidino)-2 pyrimidinylthio (N-beta-hydroxyl)acetamide (BR931) alter hepatic sex steroid metabolism and receptor expression during induction of hepatic hyperplasia and hepatocellular carcinoma (HCC) in rats. The aim of this study was to identify metabolic changes associated with cell growth during hyperplasia and HCC. METHODS: Hepatic hyperplasia was induced in male rats by a diet containing DEHP and clofibrate for 3-60 days. HCC was induced by feeding a diet containing BR931, a more potent hepatocarcinogen, for 10 months. RESULTS: Cholesterol biosynthesis was depressed in hyperplastic livers but increased in HCC. Glucose-6-phosphate dehydrogenase (G6PD) activity was inhibited in hyperplastic liver as well as in HCC, whereas malic enzyme activity increased severalfold. Protein and messenger RNA (mRNA) levels for both G6PD and malic enzyme increased in hyperplastic livers and HCC. mRNA levels for 3-hydroxy-3-methylglutaryl-coenzyme A reductase decreased in hyperplasia and increased in HCC, whereas low-density lipoprotein receptor mRNA increased in hyperplasia and decreased in HCC. CONCLUSIONS: Neoplastic cells acquire a growth advantage by their capacity to synthesize cholesterol and obtain reduced nicotinamide adenine dinucleotide phosphate by the malic enzyme pathway when G6PD activity is inhibited by peroxisome proliferators. PMID- 9207285 TI - Evidence for an ATP-dependent bile acid transport protein other than the canalicular liver ecto-ATPase in rats. AB - BACKGROUND & AIMS: Canalicular secretion is rate limiting in overall blood-to bile transport of bile acids. Studies using transfected cells have implicated the canalicular ecto-adenosine triphosphatase (ecto-ATPase) in adenosine triphosphate (ATP)-dependent bile acid transport. However, the structural features of this ecto-ATPase are not those anticipated for an in-to-out ATP-dependent transporter. The aim of this study was to explore the possible existence of an ATP-dependent bile acid transport mechanism distinct from ecto-ATPase. METHODS: Bile acid transport activity and ecto-ATPase expression were analyzed in primary rat hepatocytes, rat hepatoma HTC cells, and specially adapted HTC (HTC-R) cells using plasma membrane vesicles and Northern blot, slot blot, ribonuclease protection assay, and Western blot analyses. RESULTS: Plasma membranes isolated from HTC-R cells exhibited ATP-dependent taurocholate transport, which was many fold greater than that in HTC cells. Hepatocytes showed the highest transport rates. Protein and RNA analyses showed very low expression of ecto-ATPase in HTC and HTC-R cells compared with hepatocytes. There was no difference between the two cell types at both the RNA and protein level. CONCLUSIONS: These findings show the presence in HTC-R cells and, apparently in hepatocytes, of one or more proteins other than the ecto-ATPase that mediate ATP-dependent transport of bile acids. PMID- 9207286 TI - The rat canalicular conjugate export pump (Mrp2) is down-regulated in intrahepatic and obstructive cholestasis. AB - BACKGROUND & AIMS: The excretion of various organic anions into bile is mediated by an adenosine triphosphate-dependent conjugate export pump, which has been identified as the canalicular isoform of the multidrug resistance protein (Mrp2). Mrp2 function is impaired in various experimental models of intrahepatic and obstructive cholestasis, but the underlying molecular mechanisms are unclear. The aim of this study was to investigate these molecular mechanisms. METHODS: The effects of endotoxin, ethinylestradiol, and common bile duct ligation (CBDL) on Mrp2 protein, messenger RNA (mRNA) expression, and Mrp2 tissue localization were determined in rat livers by Northern blotting, Western analysis, and tissue immunofluorescence. To assess whether changes were specific for Mrp2, we also examined the expression of canalicular ecto-adenosine triphosphatase (ecto ATPase) and mdr P-glycoproteins (P-gp). RESULTS: All three cholestatic models resulted in a marked decrease in Mrp2 protein (P < 0.01) and its tissue localization at the canalicular membrane. Mrp2 mRNA levels diminished profoundly after endotoxin (P < 0.0005) and CBDL (P < 0.05), but did not change after ethinylestradiol. In contrast to Mrp2, protein expression of ecto-ATPase and P-gp remained unchanged in endotoxin- and ethinylestradiol-treated animals, whereas P gp levels increased after CBDL (P < 0.05). CONCLUSIONS: Down-regulation of Mrp2 expression may explain impaired biliary excretion of amphiphilic anionic conjugates in these models of cholestasis. PMID- 9207287 TI - Plasma membrane hydroxyethyl radical adducts cause antibody-dependent cytotoxicity in rat hepatocytes exposed to alcohol. AB - BACKGROUND & AIMS: We reported previously that patients with alcoholic liver disease (ALD) have circulating immunoglobulins reacting with cytochrome P4502E1 (CYP2E1) complexed with hydroxyethyl free radicals. The aim of this study was to investigate whether hydroxyethyl radical adducts are present on the plasma membranes of ethanol-treated hepatocytes and their role in antibody-dependent cytotoxicity. METHODS: Immunofluorescence confocal laser microscopy, Western blotting, and antibody-dependent cell-mediated cytotoxicity assay were used. RESULTS: Isolated rat hepatocytes incubated in vitro with ethanol or obtained from ethanol-treated animals showed strong surface fluorescence when exposed to rabbit anti-hydroxyethyl radical serum or sera from patients with ALD. No surface fluorescence was evident on control hepatocytes or after scavenging hydroxyethyl radicals with 4-pyridyl-1-oxide-t-butyl nitrone. The presence of CYP2E1 hydroxyethyl radical adducts on hepatocyte plasma membranes was shown by Western blot and by immunofluorescence using double staining for human and rabbit anti CYP2E1 immunoglobulin G. Cytotoxicity was observed in ethanol-treated hepatocytes incubated with immunoglobulin G from patients with ALD and normal human blood mononuclear cells. This effect was blocked by preabsorbing the sera with human albumin complexed with hydroxyethyl radicals, which also eliminated the antibody reaction with the plasma membranes. CONCLUSIONS: Hydroxyethyl radicals bound to CYP2E1 on hepatocyte plasma membranes can target immune reactions triggered by alcohol abuse. PMID- 9207288 TI - Induction of neutrophil-attracting chemokines in transforming rat hepatic stellate cells. AB - BACKGROUND & AIMS: Hepatic stellate cells (HSCs) play a key role in the pathogenesis of liver fibrosis. Immigrating leukocytes can potentiate the progression of liver fibrosis by release of fibrogenic mediators and cytotoxic actions. The inducible production of neutrophil chemotactic activities in HSCs was investigated to understand the underlying mechanisms responsible for the attraction of leukocytes in the pathogenesis of liver fibrosis. METHODS: Cultured HSCs of different transformation grades and after transformation to myofibroblasts (MFBs) were stimulated with tumor necrosis factor (TNF)-alpha and lipopolysaccharide (LPS), respectively. Induced leukocyte chemotactic activities were evaluated by chemotaxis assays, enzyme-linked immunosorbent assay, and Northern blot analysis. RESULTS: A transformation grade-dependent differential responsiveness of HSCs and MFBs was observed. TNF-alpha-inducible production of chemotactic mediators increased substantially with advancing transformation. Only transformed MFBs were LPS responsive. Macrophage inflammatory protein 2 was identified as one of the inducible chemokines. CONCLUSIONS: The results suggest that chemokines play an important role in the pathogenesis of liver fibrosis. Proinflammatory cytokines can initiate the production of chemotactic activities. The more HSCs are transformed to MFBs, e.g., by chronic injury, the more sensitive the cells become to LPS, which may lead to a vicious circle of enhanced fibrogenic and chemotactic mediator production. PMID- 9207290 TI - Extracellular matrix is reduced by inhibition of transforming growth factor beta1 in pancreatitis in the rat. AB - BACKGROUND & AIMS: Regeneration from cerulein-induced pancreatitis is accompanied by a transient synthesis and deposition of extracellular matrix components in the rat pancreas. The pleiotropic transforming growth factor (TGF)-beta1 has been suggested to regulate extracellular matrix remodeling during regeneration from acute pancreatitis. The present study was designed to verify this hypothesis by investigating the effect of TGF-beta1 inhibition. METHODS: Experimental acute pancreatitis was induced in rats by supramaximal doses of cerulein. The biological activity of TGF-beta1 was inhibited by injections of neutralizing TGF beta1 antibody. Changes in the content of extracellular matrix proteins, TGFs, and their messenger RNA concentrations were monitored. RESULTS: TGF-beta1 expression in pancreatic cells was suppressed after induction of acute pancreatitis by the application of neutralizing TGF-beta1 antibody. Immunochemical analysis showed a clear reduction of extracellular matrix formation during the regeneration of the pancreas in antibody-treated animals. The hydroxyproline content and the concentration of collagen types I and III, fibronectin on protein, and messenger RNA level were significantly reduced in the pancreas of treated animals. CONCLUSIONS: These results provide evidence that TGF beta1 is involved in the regulation of extracellular matrix remodeling in the rat pancreas during regeneration from acute pancreatitis. PMID- 9207289 TI - Pancreatic injury in rats induced by fatty acid ethyl ester, a nonoxidative metabolite of alcohol. AB - BACKGROUND & AIMS: The mechanism by which alcohol injures the pancreas remains unknown. Alcohol-intoxicated humans have high levels of fatty acid ethyl esters (FAEEs), nonoxidative products of ethanol metabolism, in blood, pancreas, and liver. The aims of this study were to determine whether FAEEs are toxic to the pancreas in vivo and, if so, to assess whether this injury is specific to the pancreas and to compare it to the injury observed in acute pancreatitis. METHODS: FAEEs were infused into Sprague-Dawley rats. Levels of FAEEs in plasma and pancreas were measured, and pancreatic injury was assessed during a 48-hour period for edema formation and ectopic trypsinogen activation and by light and electron microscopy. RESULTS: FAEEs induced highly significant increases in pancreatic edema, pancreatic trypsinogen activation, and vacuolization of acinar cells. These findings were specific to the pancreas and were not found in liver, lung, myocardium, skeletal muscle, or subcutaneous fat. CONCLUSIONS: FAEEs at concentrations found in human plasma produce a pancreatitis-like injury in rats, providing direct evidence that FAEEs can produce organ-specific toxicity. Thus, FAEEs may contribute to acute alcohol-induced damage to the pancreas. PMID- 9207291 TI - Cerulein-induced in vitro activation of trypsinogen in rat pancreatic acini is mediated by cathepsin B. AB - BACKGROUND & AIMS: One of the central, unresolved issues in the pathogenesis of acute pancreatitis is the uncertainty regarding the mechanisms responsible for the premature intrapancreatic activation of digestive enzyme zymogens. The aim of the current study was to develop and characterize an in vitro system that might mimic the events leading to trypsinogen activation within the pancreas during pancreatitis. METHODS: Activation of trypsinogen in response to stimulation with cerulein was quantitated in isolated rat pancreatic acini. RESULTS: Activation of trypsinogen was detected within 10 minutes of exposing isolated rat pancreatic acini, in Ca2+-containing buffer, to a supramaximally stimulating concentration of cerulein in vitro. Complete inhibition of pancreatic cathepsin B activity with E-64d, a specific, potent and irreversible cathepsin B inhibitor, prevents cerulein-induced in vitro trypsinogen activation. CONCLUSIONS: In vitro activation of trypsinogen can be detected when pancreatic acini are exposed to a supramaximally stimulating dose of cerulein. The results using this in vitro system support the hypothesis that the appearance of active trypsin within the pancreas during the early stages of cerulein-induced pancreatitis reflects activation of trypsinogen by the lysosomal hydrolase cathepsin B. PMID- 9207292 TI - Bombesin stimulates bicarbonate secretion from rat cholangiocytes: implications for neural regulation of bile secretion. AB - BACKGROUND & AIMS: Bombesin is a neuropeptide with many biological functions and is known to stimulate bile secretion. The aim of this study was to determine the role of bombesin in bile secretion and its site of action. METHODS: The effects of bombesin on bile secretion were examined using isolated perfused rat livers, hepatocyte couplets, and isolated bile duct units (IBDU) from rat liver. RESULTS: Bombesin (100 nmol/L) increased bile pH, bicarbonate concentration, and output in isolated perfused rat livers from both normal and 2-week bile duct-ligated rats, although bile flow increased only in the latter model. Bombesin (10-100 nmol/L) also had no effect on canalicular bile secretion in isolated hepatocyte couplets. However, bombesin produced a dose-dependent increase in secretion in IBDU, which was inhibited almost completely by a specific bombesin receptor inhibitor, [Tyr4, D-Phe12]-bombesin (1 micromol/L). This bombesin (10 nmol/L)-stimulated secretion in IBDU was accompanied by an increase in luminal pH and was dependent on bicarbonate and chloride in the medium. Somatostatin but not substance P inhibited the bombesin response. CONCLUSIONS: Neuropeptides such as bombesin can directly stimulate fluid and bicarbonate secretion at the level of cholangiocytes, suggesting that neuropeptides play an important regulatory role in biliary transport and secretion. PMID- 9207293 TI - Leiden factor V mutation in four patients with small bowel infarctions. AB - The Leiden factor V mutation is observed in 20% of unexplained lower limb venous thromboses and involves substitution of the arginine residue at position 506 by glutamine (R506Q). It is known to decrease the anticoagulant activity of activated protein C. This case report describes 4 cases of small bowel infarction (SBI) associated with the presence of this mutation. Two cases of arterial and 2 cases of venous SBI were observed. Extensive assessment excluded the usual causes of SBI and plasma hypercoagulation syndrome (antithrombin III, protein C, and protein S deficiency and myeloproliferative syndrome). An abnormal resistance to activated protein C was observed. Molecular analysis consisting of polymerase chain reaction amplification and digestion with MnlI showed that 2 patients were heterozygous and 2 were homozygous for the R506Q mutation. Despite familial history of thrombosis in only 1 patient, first- and second-degree relatives of 2 patients also had the presence of the mutation. Examination for the presence of abnormal resistance to activated protein C should be part of the etiological assessment of SBI. Its presence may warrant consideration of long-term anticoagulant therapy, especially for patients with shortened small bowel who are treated by home parenteral nutrition with deep venous access. PMID- 9207294 TI - The clinical correlates of a 3' truncating mutation (codons 1982-1983) in the adenomatous polyposis coli gene. AB - Familial adenomatous polyposis (FAP) is caused by mutations in the adenomatous polyposis coli (APC) gene, and different mutations may produce different clinical pictures. Most mutations occur in the 5' half of the gene, and mutations toward the 3' end are rare. The aim of this study was to document the phenotypes in a family with a truncating mutation at codons 1982-1983, one of the most 3' mutations on record. Colonic polyps in this family were much less numerous, and their growth was delayed compared with the classical FAP picture, and malignant degeneration occurred considerably later. Two individuals had sparse colonic but profuse gastric fundic gland polyposis. Gardner's syndrome stigmata were variable, and a desmoid tumor was recorded in 1 person. PMID- 9207295 TI - The oligopeptide transporter (Pept-1) in human intestine: biology and function. AB - The oligopeptide transporter (Pept-1), which is located in the intestinal brush border membrane, provides a major mechanism for protein absorption in the human intestine. Expression cloning of the gene encoding Pept-1 has predicted a 78,810 kilodalton protein consisting of 708 amino acid residues and possessing 12 putative membrane-spanning domains. The characterization of its function by in vivo and in vitro studies has shown that (1) it transports dipeptides and tripeptides but not free amino acids or peptides with more than three amino acid residues, and (2) its driving force for uphill transport requires proton binding and presence of an inside-negative membrane potential. There has also been cloning of a membrane protein (HPT-1) which appears to be associated with the oligopeptide transporter. However, the nature of association has not yet been determined. A human intestinal cell line (Caco-2), which expresses Pept-1, has been used to investigate the effects of metabolic and pathological factors on dipeptide transport. These studies suggest that the insulin stimulates dipeptide transport by increasing membrane insertion of oligopeptide transporter from a preformed cytoplasmic pool, and cholera toxin decreases dipeptide transport by inhibiting the activity of Pept-1 through an increase in the intracellular concentration of adenosine 3',5'-cyclic monophosphate. Lastly, Pept-1 seems to play important roles in nutritional and pharmacological therapies; for example, it has allowed the use of oligopeptides as a source of nitrogen for enteral feeding and the use of oral route for delivery of peptidomimetic drugs such as beta-lactam antibiotics. PMID- 9207296 TI - American Gastroenterological Association. Gallstone disease: physicochemical research joins empirical surgery. PMID- 9207297 TI - Fas and the immunologically underprivileged intestine. PMID- 9207298 TI - Muddying the water: Wilson's disease challenges will not soon disappear. PMID- 9207299 TI - Overcoming obstacles to the diagnosis of Wilson's disease. PMID- 9207300 TI - Antibodies against ethanol-derived protein adducts: pathogenic implications. PMID- 9207301 TI - The drinker's pancreas: molecular mechanisms emerge. PMID- 9207302 TI - Why are people still dying of colorectal cancer when the data indicate we should be able to prevent it? PMID- 9207303 TI - Correlating clinical outcome with molecular events in colorectal cancer. PMID- 9207304 TI - Host factors in Helicobacter infection. PMID- 9207305 TI - Colorectal cancer screening: is the gain worth the pain? PMID- 9207306 TI - The pig as an ulcer model. PMID- 9207307 TI - The emerging role of gemcitabine in lung cancer. PMID- 9207308 TI - Overview of the randomized phase III trials in non-small cell lung cancer in North America. AB - Lung cancer is the leading cause of deaths due to cancer in the United States. Although surgery can be curative for the small group of patients with early stage disease, the majority of patients present with advanced disease, for which treatment is ineffective, resulting in a 5-year overall survival rate of only 13%. Research to discover and evaluate new treatment strategies that will result in a meaningful survival benefit for patients with lung cancer is constantly ongoing. However, the process is cumbersome and requires stringent evaluation of the candidate treatment during each phase of testing. The final step in this long process is to validate the benefit of a new treatment by comparing it with the current standard treatment in the setting of a randomized phase III clinical trial. The new promising treatments for non-small cell lung cancer that are currently in phase III testing in North America are reviewed, and upcoming phase III trials are discussed. PMID- 9207309 TI - Gemcitabine: symptomatic benefit in advanced non-small cell lung cancer. AB - Recent studies have indicated that chemotherapy not only provides some survival benefit, but also reduces tumor-related symptoms and improves the performance status of patients receiving chemotherapy. Data from single-agent gemcitabine studies demonstrate improvements in a range of tumor symptoms, including cough, hemoptysis, pain, dyspnea, and anorexia, as well as increases in performance status. Indeed, more patients benefit from gemcitabine chemotherapy than suggested by the objective response rate. Surveys also have shown that patients are much more likely to accept chemotherapy for what is perceived by health care professionals as potentially small benefits. Gemcitabine has a role in the palliative treatment of patients with advanced non-small cell lung cancer. PMID- 9207310 TI - A randomized study of gemcitabine monotherapy versus etoposide/cisplatin in the treatment of locally advanced or metastatic non-small cell lung cancer. AB - This randomized, multinational, multicenter study was designed to determine the response rate of gemcitabine monotherapy and cisplatin/etoposide combination therapy in chemotherapy-naive patients with advanced, recurrent, and/or metastatic non-small cell lung cancer (stage IIIA [if inoperable], IIIB, or IV). One group of patients received gemcitabine 1,000 mg/m2 intravenously once a week for 3 weeks (days 1, 8, and 15) followed by a 1-week rest period. The second group received cisplatin 100 mg/m2 intravenously on day 1 of each 28-day cycle in combination with etoposide 100 mg/m2, administered on days 1, 2, and 3 following the cisplatin infusion. Each patient was allowed to remain on study up to a maximum of six cycles. The planned interim analysis was based on the 117 patients in the study, 116 of whom were randomized up until November 20, 1995. The efficacy analysis was performed on the 107 patients who had data from a minimum of two cycles, whereas the safety analysis was based on data from all 116 randomized patients. In the gemcitabine arm there were 10 of 52 (19%) partial responders; in the cisplatin/etoposide arm there were four (7%) of 54 partial responders. There was a statistically significant difference in the response rates between the two arms, with a 95% confidence interval of 0.6% to 32.1% (P = .040). The median time to progressive disease was 4.2 months for gemcitabine patients and 3.7 months for cisplatin/etoposide patients. There was significantly more alopecia and nausea and vomiting in the cisplatin/etoposide arm compared with the gemcitabine arm, as well as two cases of neutropenic sepsis in the cisplatin/etoposide arm. These data indicate that single-agent gemcitabine is at least as effective as the combination of cisplatin/etoposide in the treatment of advanced non-small cell lung cancer and has an improved safety profile. PMID- 9207311 TI - Weekly gemcitabine and monthly cisplatin for advanced non-small cell lung carcinoma. AB - Most chemotherapy for non-small cell lung cancer (NSCLC) currently includes combination chemotherapy based on cisplatin. Gemcitabine is a nucleoside analog with demonstrated activity against NSCLC, yet it has low toxicity. This phase II study was designed to examine the efficacy of a combination chemotherapy regimen consisting of gemcitabine followed by cisplatin. The patient population comprised 53 patients with pathologically confirmed locally advanced or metastatic NSCLC. Gemcitabine 1,000 mg/m2 was administered on days 1, 8, and 15 and cisplatin 100 mg/m2 was given on day 15. Chemotherapy was administered every 28 days. Of the 50 patients evaluable for response, there were two complete responses (4%) and 24 partial responses (48%). The median duration of response was 8.5 months, median survival was 13 months, and the 1-year survival rate was 61%. The regimen was generally well tolerated. World Health Organization grade 3 leukopenia occurred in 28.8% of patients, while grade 3 and 4 neutropenia occurred in 38.8% and 19.2% of patients, respectively. Grade 3 and 4 thrombocytopenia was seen in 13.3% and 7.7% of patients, and grade 3 and 4 anemia occurred in 11.5% and 1.9% of patients, respectively. Alopecia and oral toxicity was mild, although most patients experienced mild nausea and vomiting. Relatively few patients required dose modifications for any of the three weekly doses of chemotherapy. We conclude that the combination of gemcitabine and cisplatin is an effective regimen for NSCLC, resulting in high response and survival rates. Additional prospective randomized studies with other cisplatin-based combination chemotherapy regimens are warranted. PMID- 9207312 TI - Phase II trial of gemcitabine plus cisplatin in non-small cell lung cancer: a Hoosier Oncology Group study. AB - Although non-small cell lung cancer accounts for a majority of lung cancer cases in the United States, overall response rates of only 20% have been obtained with cisplatin-based therapy. However, new agents such as the nucleoside analog gemcitabine have demonstrated single-agent response rates of approximately 20%. The synergistic growth inhibition of several tumor cell lines seen in vitro with cisplatin and gemcitabine has led to clinical trials incorporating this drug combination. Recent phase I trials revealed that administration of gemcitabine 1,000 mg/m2 weekly x 3, with cisplatin 100 mg/m2 on day 15, every 28 days, was well tolerated. This regimen was used in the present Hoosier Oncology Group phase II trial of patients with advanced non-small cell lung cancer, except that cisplatin was administered on day 1. Five patients had stage IIIB and 25 had stage IV disease, with a mean Karnofsky performance status of 90. Toxicity observed was primarily hematologic, notably, granulocytopenia and thrombocytopenia. Responses were seen in 10 of 27 patients, for an overall response rate of 37%. Median overall survival was 8.4 months; it was 10.2 months for patients with stage IV disease. These favorable results and manageable side effects suggest that future trials incorporating gemcitabine and cisplatin are indicated. PMID- 9207313 TI - Phase II trial of gemcitabine and weekly cisplatin for advanced non-small cell lung cancer. AB - Single-agent gemcitabine, when given in doses of > or = 1,250 mg/m2 weekly x 3 with a 1-week break, induces responses in approximately 20% of untreated patients with non-small cell lung cancer. This phase II study was undertaken to determine the efficacy of weekly administration of gemcitabine 1,500 mg/m2 combined with cisplatin 30 mg/m2 x 3 with a rest period of 1 week. Patients younger than 75 years were eligible if they had stage III/IV non-small cell lung cancer, a life expectancy > or = 12 weeks, hemoglobin > or = 10 g/dL, absolute granulocyte count > or = 10(9)/L, platelets > or = 100 x 10(9)/L, hepatic enzymes no more than three times the upper limit of normal, and serum creatinine < or = 130 micromol/L. There were 22 men and 18 women, with a median age of 60 years; 35 had a performance status of 0 or 1. Pathology included adenocarcinoma in 22 patients, squamous cell carcinoma in nine, large cell carcinoma in seven, and mixed non small cell lung cancer in two. Six patients had stage III and 34 had stage IV tumors. Of the 39 patients eligible for response evaluation, partial remission was seen in 10, for an overall response rate of 26% (95% confidence interval, 12% to 41%). The median duration of response was 19 weeks (range, 7 to 32+ weeks). Grade 3/4 anemia was seen in 11 patients, and 21 patients required red blood cell transfusions. Grade 3/4 neutropenia occurred in 22 patients and grade 3/4 thrombocytopenia in 21 patients. One patient experienced febrile neutropenia Hematologic toxicity, particularly thrombocytopenia, was cumulative over time. Nonhematologic toxicity was modest, but one patient stopped therapy because of a grade 2 skin rash and one stopped because of a grade 4 pulmonary toxicity, both of which were thought to be related to gemcitabine. The modest activity of weekly gemcitabine and weekly cisplatin seen in this trial does not suggest in vivo synergy for these two agents as administered using this schedule and these doses. PMID- 9207314 TI - Rationale of a phase III study comparing a standard cisplatin regimen (mitomycin/ifosfamide/cisplatin) with cisplatin and gemcitabine in non-small cell lung cancer. AB - Cisplatin-based combination chemotherapy is currently considered the most active treatment for advanced non-small cell lung cancer (NSCLC). A recent meta-analysis of eight randomized trials comparing supportive care versus supportive care plus cisplatin-based chemotherapy showed a small (10%) but significant survival benefit at 1 year. However, there is no consensus on a specific reference regimen for NSCLC. Our previous experience comparing cisplatin/etoposide (PE) with mitomycin/ifosfamide/cisplatin (MIC) demonstrated a significantly better response rate (40% v 23%) and survival advantage of the three-drug MIC combination versus PE. Among the new drugs, the nucleoside analogue gemcitabine produced response rates of approximately 20% in advanced NSCLC. In phase I/II studies, response rates as high as 54% and median survival of up to 13 months were seen when gemcitabine was combined with cisplatin. Therefore, a phase III study was planned to evaluate the gemcitabine/cisplatin combination versus the MIC regimen. This multicenter trial is currently ongoing in Italy. PMID- 9207315 TI - A phase II trial of gemcitabine and ifosfamide in non-small cell lung cancer. AB - The novel nucleoside agent gemcitabine has demonstrated antitumor activity against a variety of solid tumors and is associated with low toxicity. A phase I trial in Germany of gemcitabine combined with the alkylating agent ifosfamide has shown encouraging activity against non-small cell lung cancer (NSCLC). The efficacy and toxicity of this combination was further evaluated in a phase II trial of chemotherapy-naive patients with NSCLC (mostly stage IV disease). Gemcitabine was administered at a dose of 1,000 mg/m2 on days 1, 8, and 15 followed by a 1-week rest, while ifosfamide was given at a dose of 1,500 mg/m2 on day 8 and days 9 through 12. Fifty-one of 56 patients were evaluable for response. Eleven partial responses were seen, for an overall objective response rate of 22%. The 6- and 9-month survival rates are 62% and 41%, respectively. Grade 3 and 4 neutropenia occurred in 35.8% and 24.5% of patients, respectively, but the incidence of infection was low. These results indicate that the combination of gemcitabine and ifosfamide is active against NSCLC and has a mild toxicity profile, and suggest that further evaluation of this combination is warranted. PMID- 9207317 TI - Investigations of new drugs in combination with cisplatin in stage III non-small cell lung cancer. AB - Chemotherapy has been shown to prolong the survival of patients with stage III non-small cell lung cancer. In particular, cisplatin and vinblastine administered before definitive radiotherapy results in a significant prolongation of median survival and increases the 2-year survival rate. Concomitant chemoradiotherapy also holds promise. Recently, several new active agents and combinations have been described for the treatment of stage III disease. The integration of these drugs and novel regimens into treatment plans for stage III non-small cell lung cancer is of great interest and a focus of current clinical investigations. Early data from these clinical trials are reviewed here. PMID- 9207316 TI - Gemcitabine plus radiotherapy for non-small cell lung cancer. AB - Radiosensitization promises to improve local control in patients with locally advanced non-small cell lung cancer (NSCLC). The nucleoside analogue gemcitabine, which is not only a potent radiosensitizer but also has significant antitumor activity in this disease, is an important candidate for such a strategy. This report describes the design of a clinical phase I study for treatment of patients with locally advanced NSCLC using gemcitabine and radiotherapy. It also discusses some of the practical constraints that must be taken into consideration when evaluating this type of combined modality strategy in the clinic. PMID- 9207319 TI - A piece of my mind. The chief complaint. PMID- 9207318 TI - Systemic investigational therapies as adjuvants to surgery in patients with operable lung cancer. AB - Early resection of lung cancers confined to the chest remains the best means of curing patients. Nonetheless, cure rates for resected patients are less than 50% overall, primarily due to the presence of occult distant metastases at the time of resection that subsequently manifest as relapse at distant sites. In approximately 20% of patients, local failure occurs within the chest. Chest radiotherapy reduces and nearly eliminates local failure, but does not improve survival or cure rates. Effective systemic chemotherapy is the cornerstone to improving survival and cure rates for early stage lung cancer. Recent studies showed that the preoperative use of cisplatin-based chemotherapy improved survival and 3-year disease-free survival in stage IIIA non-small cell lung cancer. New drugs such as paclitaxel and gemcitabine appear to improve survival in stages IIIB and IV non-small cell lung cancer. These agents will be tested in the surgical setting in the near future. PMID- 9207320 TI - Taking asthma seriously. PMID- 9207321 TI - Psychotherapy reduces disability, saves money. PMID- 9207322 TI - Put human cloning on hold, say bioethicists. PMID- 9207323 TI - From the Food and Drug Administration. PMID- 9207324 TI - From the Centers for Disease Control and Prevention. Amanita phalloides mushroom poisoning--northern California, January 1997. PMID- 9207325 TI - From the Centers for Disease Control and Prevention. Cigar smoking among teenagers--United States, Massachusetts, and New York, 1996. PMID- 9207326 TI - Professionalism vs commercialism in managed care: the need for a national council on medical care. PMID- 9207327 TI - Professionalism vs commercialism in managed care: the need for a national council on medical care. PMID- 9207328 TI - Professionalism vs commercialism in managed care: the need for a national council on medical care. PMID- 9207329 TI - Professionalism vs commercialism in managed care: the need for a national council on medical care. PMID- 9207330 TI - Is residency training really harmful? PMID- 9207331 TI - Use of optical immunoassay to diagnose streptococcal pharyngitis. PMID- 9207332 TI - Medicaid coverage for young women and children: whose welfare is at stake? PMID- 9207333 TI - Identifying the causative organism in patients with ventilator-associated pneumonia. PMID- 9207334 TI - Benzodiazepine use and the risk of motor vehicle crash in the elderly. AB - CONTEXT: Benzodiazepines, used by a sizable number of the elderly population, may affect the ability to drive and thus increase the risk of a motor vehicle crash. Epidemiologic studies of this question have produced inconsistent results that may be due to the different effects of long- and short-half-life benzodiazepines and variations in their duration of use. OBJECTIVE: To determine whether the use of benzodiazepines of either long- or short-elimination half-life is associated with the risk of injurious motor vehicle crash in the elderly. DESIGN AND SETTING: Nested case-control design within a cohort of 224,734 drivers from the Canadian province of Quebec, aged 67 to 84 years, followed up from 1990 to 1993. Computerized data for the study were obtained from provincial driver's license files, police reports of injurious crashes, and health insurance records. PATIENTS: We identified all 5579 drivers involved in an injurious crash (cases) and a random sample of 10 controls per case selected from a subcohort of 13,256 subjects. MAIN OUTCOME: Involvement of a cohort member as a driver in a motor vehicle crash in which at least 1 person (not necessarily the driver) sustained bodily injury. RESULTS: The adjusted rate ratio of crash involvement within the first week of long-half-life benzodiazepine use was 1.45 (95% confidence interval [CI], 1.04-2.03). The rate ratio for continuous use of longer duration up to 1 year was slightly lower but remained significant (rate ratio, 1.26; 95% CI, 1.09 1.45). In contrast, there was no increased risk after the initiation of treatment with short-half-life benzodiazepines (rate ratio, 1.04; 95% CI, 0.81-1.34) or with their continued use (rate ratio, 0.91; 95% CI, 0.82-1.01). CONCLUSIONS: Brief or extended periods of exposure to long-half-life benzodiazepines are associated with an increased risk of motor vehicle crash involvement in the elderly population. There is no such elevated risk for short-half-life benzodiazepines. PMID- 9207335 TI - Medical outcomes and antimicrobial costs with the use of the American Thoracic Society guidelines for outpatients with community-acquired pneumonia. AB - CONTEXT: The American Thoracic Society (ATS) published guidelines based on expert opinion and published data--but not clinically derived or validated--for treating adult outpatients with community-acquired pneumonia. OBJECTIVE: To compare medical outcomes and antimicrobial costs for patients whose antimicrobial therapy was consistent or inconsistent with ATS guidelines. DESIGN: Multicenter, prospective cohort study. SETTING: Emergency departments, medical clinics, and practitioner offices affiliated with 3 university hospitals, 1 community teaching hospital, and 1 health maintenance organization. PARTICIPANTS: A total of 864 immunocompetent, adult outpatients with community-acquired pneumonia: 546 aged 60 years or younger with no comorbidity and 318 older than 60 years or with 1 comorbidity or more. MAIN OUTCOME MEASURES: Patients' antimicrobial therapy was classified as being consistent or inconsistent with the ATS guidelines. Mortality, subsequent hospitalization, medical complications, symptom resolution, return to work and usual activities, health-related quality of life, and antimicrobial costs were compared among those treated consistently or inconsistently with the guidelines. RESULTS: Outpatients aged 60 years or younger with no comorbidity who were prescribed therapy consistent with ATS guidelines (ie, erythromycin with some exceptions) had 3-fold lower antimicrobial costs ($5.43 vs $18.51; P<.001) and no significant differences in medical outcomes. Outpatients older than 60 years or with 1 comorbidity or more who were prescribed therapy consistent with ATS guidelines (ie, second-generation cephalosporin, sulfamethoxazole-trimethoprim, or beta-lactam and beta-lactamase inhibitor with or without a macrolide) had 10-fold higher antimicrobial costs ($73.50 vs $7.50; P<.001); despite trends toward higher mortality and subsequent hospitalization, no significant differences in medical outcomes were observed. CONCLUSION: Our findings support the use of erythromycin as recommended by the ATS guidelines for outpatients aged 60 years or younger with no comorbidity. Although the antimicrobial therapy recommended in outpatients older than 60 years or with 1 comorbidity or more is more costly, this observational study provides no evidence of improved medical outcomes in the small subgroup who received ATS guideline recommended therapy. PMID- 9207336 TI - Antihypertensives and the risk of serious hypoglycemia in older persons using insulin or sulfonylureas. AB - CONTEXT: Beta-Blockers and angiotensin-converting enzyme (ACE) inhibitors are effective antihypertensive agents for patients with diabetes mellitus. However, beta-blockers attenuate some components of the autonomic response to hypoglycemia and could increase the risk of hypoglycemia. ACE inhibitors may increase insulin sensitivity and predispose users to hypoglycemia. OBJECTIVE: To determine whether use of cardioselective beta-blockers, nonselective beta-blockers, ACE inhibitors, thiazide diuretics, calcium channel blockers, or other antihypertensive drugs alters the risk of developing serious hypoglycemia among older persons prescribed insulin or sulfonylureas. DESIGN: Retrospective cohort study. SETTING: Tennessee Medicaid Program. PATIENTS: A total of 13,559 elderly (mean age, 78+/-7 years) Medicaid enrollees, who were prescribed insulin (n=5171, 38%) or sulfonylureas (n=8368, 62%) from 1985 through 1989. These enrollees contributed a total of 33,107 person-years of insulin or sulfonylurea use for follow-up. MEASUREMENTS: Hospitalization, emergency department admission, or death associated with hypoglycemic symptoms and a concomitant blood glucose determination of less than 2.8 mmol/L (50 mg/dL). RESULTS: We identified 598 persons with an episode of serious hypoglycemia during the study period. The rate of serious hypoglycemia was 2.01 per 100 person-years among those who were not prescribed antihypertensives. Crude rates of serious hypoglycemia were highest among users of ACE inhibitors (2.47 per 100 person-years) and lowest among users of cardioselective beta-blockers (1.23 per 100 person-years). However, when we controlled for demographic characteristics and markers of comorbidity, there was no statistically significant increase or decrease in risk of serious hypoglycemia among users of any class of antihypertensive agents compared with nonusers of antihypertensive drugs. Using nonselective beta-blockers as the reference group, each of these agents was associated with a lower, but not statistically significant, risk of hypoglycemia. CONCLUSIONS: In this population, specific antihypertensive drug therapy had little impact on the risk of hypoglycemia in older diabetic patients. Therapy should be chosen based on other considerations of safety and effectiveness. PMID- 9207337 TI - Survival after radical prostatectomy. AB - CONTEXT: The generalizability of currently available estimates of survival after radical prostatectomy is theoretically limited. OBJECTIVE: To obtain generalizable estimates of survival after radical prostatectomy. DESIGN: A population-based retrospective cohort study. SETTING: Nine regions of the United States. PATIENTS: Patients who were diagnosed with prostate cancer between 1983 and 1987 and underwent radical prostatectomy and lymph node dissection. MAIN OUTCOME MEASURES: Proportional hazards models incorporating geographical region, age, race, pathological stage, lymph node involvement, and tumor grade to identify independent correlates of disease-specific and overall survival and life table analyses to estimate 10-year survival distributions. RESULTS: A total of 3626 patients with a mean age of 65 years were included in the study; 92.6% were white, 54.2% had moderate-grade cancer, 60.4% had no extension beyond the prostate, and 91.2% had no lymph node involvement. Using San Francisco-Oakland, Calif, as a reference region, no other region was significantly associated with a risk of disease-specific or overall mortality. Older age and black race were independently associated with worse overall but not disease-specific survival. Higher grade, extension beyond the prostate, and lymph node involvement were independently associated with worse disease-specific and overall survival. Estimates of 10-year disease-specific survival ranged from 75% to 97% for patients with well-differentiated and moderately differentiated cancers and from 60% to 86% for patients with poorly differentiated cancers. CONCLUSIONS: Neither disease-specific nor overall survival varied by region, suggesting geographically uniform assessments of risk in patient selection for radical prostatectomy. Across regions, overall survival varied by patient and prostate cancer characteristics while disease-specific survival varied substantially by prostate cancer but not patient characteristics. The present analyses provide the most generalizable current estimates of survival after radical prostatectomy. PMID- 9207338 TI - The problem of quality of life in medicine. AB - The use of the term "quality of life" to encompass the values and perceptions of patients has created doubt, confusion, and misunderstanding among practitioners, researchers, policymakers, and patients. The principal reason for this state of affairs is that a clear conceptual basis for quality-of-life measures is lacking. In this article, the current rationale for quality-of-life measurement in the health field is examined, and the drawbacks of the various models being used are outlined. Our suggestion is that quality of life as an outcome could be explored more clearly (ie, defined) if quality of life were replaced with a more easily handled notion such as that of "subjective health status." However, the idea that the patient's perspective is as valid as that of the clinician when it comes to evaluating outcomes has a great deal of legitimacy and should certainly not be abandoned. PMID- 9207339 TI - The diagnostic evaluation and multidisciplinary management of neurofibromatosis 1 and neurofibromatosis 2. AB - OBJECTIVE: Neurofibromatosis 1 and neurofibromatosis 2 are autosomal dominant genetic disorders in which affected individuals develop both benign and malignant tumors at an increased frequency. Since the original National Institutes of Health Consensus Development Conference in 1987, there has been significant progress toward a more complete understanding of the molecular bases for neurofibromatosis 1 and neurofibromatosis 2. Our objective was to determine the diagnostic criteria for neurofibromatosis 1 and neurofibromatosis 2, recommendations for the care of patients and their families at diagnosis and during routine follow-up, and the role of DNA diagnostic testing in the evaluation of these disorders. DATA SOURCES: Published reports from 1966 through 1996 obtained by MEDLINE search and studies presented at national and international meetings. STUDY SELECTION: All studies were reviewed and analyzed by consensus from multiple authors. DATA EXTRACTION: Peer-reviewed published data were critically evaluated by independent extraction by multiple authors. DATA SYNTHESIS: The main results of the review were qualitative and were reviewed by neurofibromatosis clinical directors worldwide through an Internet Web site. CONCLUSIONS: On the basis of the information presented in this review, we propose a comprehensive approach to the diagnosis and treatment of individuals with neurofibromatosis 1 and neurofibromatosis 2. PMID- 9207340 TI - Acute primary HIV infection. PMID- 9207341 TI - Protease inhibitors in the homeless. PMID- 9207342 TI - Safety and mobility of the older driver: a research challenge. PMID- 9207343 TI - JC virus (JCV) genotypes in brain tissue from patients with progressive multifocal leukoencephalopathy (PML) and in urine from controls without PML: increased frequency of JCV type 2 in PML. AB - Progressive multifocal leukoencephalopathy (PML) is caused by the human polyomavirus JC (JCV), and there are at least 4 different genotypes of JCV in the United States. Type 1 strains are of European origin, whereas type 2 and 3 strains are of Asian and African origin, respectively. JCV type 4 strains are derived from a type 1/3 recombinant. In this study, the genotype distribution of JCV strains found in brain tissue or cerebrospinal fluid of 50 PML patients was compared with JCV genotypes excreted in the urine of 103 control subjects. Type determination was based on the polymerase chain reaction-amplified partial sequence of the VP1 coding gene and the noncoding region left of ori. Brain tissues from patients with PML were infected with a significantly higher proportion of JCV type 2 strains than were urine samples from the control group (P = .004). This evidence indicates a biologic difference between JCV genotypes and suggests a difference in their potential to cause PML. PMID- 9207344 TI - Immunoprecipitation and virus neutralization assays demonstrate qualitative differences between protective antibody responses to inactivated hepatitis A vaccine and passive immunization with immune globulin. AB - Antibodies to hepatitis A virus (anti-HAV) were measured in children from two separate vaccine trials (n = 70) 4 weeks after a dose of inactivated hepatitis A vaccine (VAQTA). The geometric mean titers (GMTs) of anti-HAV were 49.3 and 45.2 mIU/mL by immunoassay, while reciprocal GMTs of neutralizing anti-HAV were 6.5 and 15.0 by an 80% radioimmunofocus inhibition test (RIFIT) and 55.6 and 92.0 by antigen reduction assay (HAVARNA). The GMT of antibody detected by radioimmunoprecipitation (RIPA) was > or =401. These data establish serologic correlates of protection against disease and show that RIPA is most sensitive for detection of early vaccine-induced antibody. Sera collected from adults (n = 20) 7 days after administration of immune globulin contained similar antibody levels by immunoassay (45.1 mIU/mL) and slightly higher GMTs of neutralizing antibody (27.5 by RIFIT and 146 by HAVARNA) but negligible precipitating antibody (GMT, 5.6). These results are best explained by differences in the affinity of antibodies for virus following active versus passive immunization. PMID- 9207345 TI - Distribution of hepatitis C virus (HCV) RNA in whole blood and blood cell fractions: plasma HCV RNA analysis underestimates circulating virus load. AB - Previous experiments using a cationic surfactant to detect hepatitis C virus (HCV) RNA in whole blood (WB) suggested that WB was a more plentiful source of viral RNA than was plasma. The relative HCV RNA titers in WB, plasma, peripheral blood mononuclear cells (PBMC), neutrophils, and red blood cells (RBC)/platelets from 10 patients with chronic HCV infection were compared. WB contained significantly more HCV RNA than plasma, which contained more HCV RNA than PBMC, neutrophils, or RBC/platelets (P < .001). To determine if this increased sensitivity was clinically relevant, results of WB and plasma HCV RNA assays were compared with commercial quantitative and qualitative plasma HCV RNA assay results obtained for patients receiving interferon therapy. WB was significantly more sensitive than commercial plasma reverse transcription-polymerase chain reaction for detecting HCV RNA (P < .005). These data indicate that a significant proportion of HCV RNA in peripheral blood is not identified by standard plasma RNA detection methods. PMID- 9207346 TI - Surreptitious hepatitis C virus (HCV) infection detected in the majority of patients with cryptogenic chronic hepatitis and negative HCV antibody tests. AB - Reverse transcription-polymerase chain reaction was used to identify hepatitis C virus (HCV) RNA in peripheral whole blood (WB) and plasma samples from 15 patients with chronic, unexplained hepatitis. These patients were serologically negative for hepatitis A, B, and C and were classified as having chronic non-A, non-B, non-C hepatitis (NANBNC). HCV RNA was repeatedly detected in WB samples from 10 (67%). In contrast, plasma samples from only 5 were intermittently positive. Statistically, HCV RNA detection in WB was significantly more sensitive than in plasma. Nucleic acid hybridization and HCV genotypic analysis confirmed the specificity of the HCV RNA assay. Liver biopsies from these patients suggested histopathologic differences between HCV RNA-positive and -negative groups. These data demonstrate that HCV infection is present in patients with unexplained chronic hepatitis more frequently than previously believed. Additionally, WB HCV RNA detection is more sensitive than plasma assays in identifying antibody-negative HCV infection. PMID- 9207348 TI - Maternal factors affecting the integrity of the late gestation placental barrier to murine enterovirus infection. AB - Maternal factors that might impair the integrity of the late gestation placental barrier to enteroviruses were evaluated. Mice were inoculated with Theiler's murine encephalomyelitis virus (TMEV) on day 10-13 of gestation and sacrificed on day 16-18. Placentas and fetuses from dams with advanced age, forced daily swimming, short-term clamping of uteroplacental blood vessels, restricted dietary intake, or bacterial peritonitis were compared with tissues from TMEV-infected control mice. Increased maternal age, exercise, and malnutrition were associated with reduced fetal weight, and disturbed uteroplacental blood flow and severe malnutrition were associated with abnormal placental and fetal morphology. TMEV infection was observed sporadically by culture or in situ hybridization (or both) in fetuses from dams with interrupted uteroplacental blood flow, bacterial peritonitis, and older age but not in fetuses from control infected mice. This suggests that maternal factors, such as compromised uteroplacental blood flow, concomitant infection, and advanced age, may increase the risk of transplacental fetal infection. PMID- 9207347 TI - Prevalence of and risk factors for antibody to hepatitis E virus seroreactivity among blood donors in Northern California. AB - To evaluate antibody to hepatitis E virus (anti-HEV) seroreactivity, 5000 US blood donors were tested for anti-HEV by two EIAs: a mosaic protein assay (MPr EIA) and a recombinant protein assay (RPr-EIA). Overall, 59 (1.2%) were seroreactive by MPr-EIA and 70 (1.4%) were seroreactive by RPr-EIA. The overall concordance between tests was 98.5% (4925/5000); the concordance among reactive sera by either test was only 27% (27/102). In a case-control study, seroreactive persons were more likely than seronegative persons to have traveled to countries in which HEV is endemic (odds ratio [OR] for MPr-EIA = 4.3, P < .001; OR for RPr EIA = 2.5, P = .005), but 31% of MPr-EIA anti-HEV-reactive persons and 38% of RPr EIA anti-HEV-reactive persons had no history of international travel. These findings suggest that travelers to regions in which HEV is endemic can acquire subclinical HEV infection. The significance of anti-HEV seroreactivity among persons without an international travel history needs to be determined. PMID- 9207349 TI - Cytomegalovirus (CMV) culture results, drug resistance, and clinical outcome in patients with AIDS and CMV retinitis treated with foscarnet or ganciclovir. Studies of Ocular Complications of AIDS (SOCA) in collaboration with the AIDS Clinical Trial Group. AB - The objectives of the study were to examine the clinical significance of cytomegalovirus (CMV) culture results and drug susceptibilities in CMV isolates from patients with AIDS-related CMV retinitis. Blood and urine for CMV culture were obtained from 207 patients with newly diagnosed CMV retinitis who were enrolled in a randomized trial comparing foscarnet and ganciclovir. Culture positive rates at baseline were 45% and 71% for blood and urine, respectively. Rates decreased 3- to 10-fold after initiation of either treatment. Mortality was related to both positive baseline blood and urine cultures; adjusted relative risks were 1.97 and 2.03, respectively. Positive blood cultures at baseline were associated with more rapid retinitis progression. Drug-resistant CMV was found, over comparable follow-up periods on assigned treatment, in 4 of 8 ganciclovir assigned patients with persistent viremia and 0 of 5 foscarnet-assigned patients with persistent viremia. Results of virologic assays of blood appear to be associated with clinical outcome of CMV retinitis. PMID- 9207350 TI - A humanized antibody against human cytomegalovirus (CMV) gpUL75 (gH) for prophylaxis or treatment of CMV infections. AB - A humanized monoclonal antibody that binds to the 86-kDa glycoprotein, gpUL75 (gH), of human cytomegalovirus (CMV) has been developed. The six complementarity determining regions of the heavy and light chains of the mouse antibody HCMV16 were transferred to human antibody framework sequences and combined with human antibody constant regions to produce a complete antibody. The reshaped antibody recognized cells infected with a variety of virus strains and neutralized clinical isolates of CMV as efficiently as laboratory strains in a conventional plaque reduction assay. This antibody provides a potential agent for the prevention or treatment of CMV infections in humans. PMID- 9207351 TI - High-level resistance of cytomegalovirus to ganciclovir is associated with alterations in both the UL97 and DNA polymerase genes. AB - Mutations in both the viral phosphotransferase gene, UL97, and the DNA polymerase gene, UL54, have been shown to confer ganciclovir resistance to cytomegalovirus (CMV). Moreover, UL54 alterations have been associated with in vitro cross resistance of CMV to cidofovir. To investigate the relative significance of UL97 versus UL54 alterations in conferring antiviral resistance, phenotypic and genotypic characterization of 28 ganciclovir-resistant clinical CMV isolates was undertaken. Isolates were either low-level ganciclovir-resistant, which have ganciclovir ID50 values > or =8 microM and <30 microM and sensitivity to cidofovir, or high-level ganciclovir-resistant, which have ganciclovir ID50 values > or =30 microM and cross-resistance to cidofovir. Low-level ganciclovir resistant isolates were associated with UL97 alterations and short periods of ganciclovir treatment, while high-level ganciclovir-resistant isolates were associated with both UL97 and polymerase alterations and were cultured after extended ganciclovir therapy. PMID- 9207352 TI - Topical undecylenic acid for herpes simplex labialis: a multicenter, placebo controlled trial. AB - A multicenter, patient-initiated, double-blind, placebo-controlled trial of 15% undecylenic acid cream was conducted with 573 patients with recurrent herpes labialis. Treatment was applied 5 or 6 times daily until crusting and then thrice daily until healing. Patients were assessed daily until 48 h after crusting and then every other day until healing. Undecylenic acid significantly reduced the incidence and duration of viral shedding and the duration and severity of itching but did not increase abortive episodes or reduce times to healing, crusting, or progression of lesion size. When treatment was initiated during the prodrome, the time to crusting was reduced (P = .02) and the area under the symptom-time curve for pain and tenderness was reduced, approaching statistical significance (P = .06). Active treatment was well tolerated but caused dysgeusia and local irritation. Undecylenic acid 15% cream reduces viral shedding in recurrent herpes labialis, but clinical benefits are minimal and largely restricted to patients initiating therapy during the prodrome. PMID- 9207353 TI - Detection of human herpesvirus 8 DNA in Kaposi's sarcoma lesions and peripheral blood of human immunodeficiency virus-positive patients and correlation with serologic measurements. AB - Polymerase chain reaction (PCR) was used to examine human herpesvirus 8 (HHV-8) DNA from Kaposi's sarcoma (KS) lesions, normal skin, and peripheral blood mononuclear cells (PBMC) from human immunodeficiency virus (HIV)-infected patients who did or did not have KS. Of 9 KS biopsies, 8 were positive for five HHV-8 open-reading frames and ranged from 1 viral genome per 2.5-12.7 cells. Two putative replicative gene RNAs were detected by reverse transcription-PCR at low levels in 1 KS lesion. HHV-8 DNA was detected in 4 of 8 PBMC samples from patients with KS and in 2 of 18 PBMC samples from patients without KS. Sera were tested for reactivity with BCBL-1 cells (HHV-8 positive): High immunofluorescence antibody titers against HHV-8 lytic and latent antigens were detected in samples from KS-positive patients, and >20 polypeptides from induced BCBL-1 cells were recognized. Sera from 6 of 18 patients without KS showed low levels of antibodies against HHV-8 lytic and latent antigens. PMID- 9207354 TI - Frequent detection of Kaposi's sarcoma-associated herpesvirus (human herpesvirus 8) DNA in saliva of human immunodeficiency virus-infected men: clinical and immunologic correlates. AB - The prevalence, quantity, temporal pattern, and clinical and immunologic correlates of shedding of Kaposi's sarcoma (KS)-associated herpesvirus (KSHV; or human herpesvirus [HHV]-8) DNA in saliva were studied. KSHV DNA was detected in saliva from 18 (75%) of 24 human immunodeficiency virus (HIV)-positive patients with KS and from 1 of 1 HIV-negative patient with KS, 3 (15%) of 20 HIV-positive patients without KS, and none of 24 controls. KSHV DNA levels ranged from 10(2.4) to 10(6) copies/mL and were lower than levels for Epstein-Barr virus but comparable to those for HHV-6. Detection of KSHV DNA in saliva was not associated with oral KS or decreased peripheral blood CD4 cell counts. KSHV DNA was not detected in semen. Resistance of KSHV DNA from saliva to DNase treatment was consistent with the presence of virions. These data suggest that KSHV can replicate in the oropharynx and that salivary contact could contribute to KSHV transmission. PMID- 9207355 TI - Evaluation of sorivudine (BV-araU) versus acyclovir in the treatment of acute localized herpes zoster in human immunodeficiency virus-infected adults. The Multinational Sorivudine Study Group. AB - The clinical efficacy and safety of sorivudine as treatment for acute cutaneous zoster in human immunodeficiency virus-infected adults was compared with that of acyclovir in a double-blinded randomized study. A total of 125 patients with laboratory-confirmed zoster rash present for < or =72 h were assigned treatment with either 40 mg of sorivudine once daily or 800 mg of acyclovir five times daily, both taken orally for 7 days. Patients were assessed daily until all lesions crusted and then monthly for 6 months for postherpetic neuralgia (PHN) and for 12 months for recurrent or new episodes of zoster. Sorivudine significantly shortened the median period of new vesicle formation from 3.0 to 4.0 days (log rank P = .0001). Sorivudine was effective regardless of duration of rash before treatment. Zoster recurrences and new episodes were experienced by fewer patients assigned sorivudine (11%) than acyclovir (26%, P = .037). No differences were seen in incidence, severity, or duration of either acute neuritis or PHN. Both treatments were well tolerated. PMID- 9207356 TI - Incidence and prognostic significance of symptomatic primary human immunodeficiency virus type 1 infection in homosexual men. AB - To investigate the incidence of symptomatic primary human immunodeficiency virus type 1 (HIV-1) infection and its prognostic significance for HIV-1 disease progression, data for 328 homosexual men from four cohort studies were evaluated. Rates of diarrhea, fever, night sweats, cough, and fatigue prior to, during, and after seroconversion were compared by use of Poisson regression, and the prognostic significance of these symptoms was evaluated with survival methods. The incidence of all symptoms was elevated during seroconversion; however, only fever was associated with faster disease progression. Seven or more days of fever was reported by 13.8% of subjects; half of them developed AIDS within 6 years, whereas only one-fourth of the men without fever developed AIDS within 6 years. In addition, fever was the only symptom associated with shortened survival and increased CD4 cell loss. Persons experiencing prolonged periods of fever during seroconversion should therefore be considered for early treatment, including prophylaxis against opportunistic infections and combinations of antiretroviral drugs. PMID- 9207357 TI - Human immunodeficiency virus type 1-specific cytotoxic T lymphocytes (CTL), virus load, and CD4 T cell loss: evidence supporting a protective role for CTL in vivo. AB - The relationships between primary human immunodeficiency virus type 1 (HIV-1) Gag specific cytotoxic T lymphocyte (CTL) frequency, virus load, and CD4 T cell loss were evaluated in a group of 46 HIV-1-infected persons with hemophilia. Freshly isolated peripheral blood mononuclear cells in limiting dilution assays were used to measure HIV-1 Gag-specific CTL frequencies. Concurrent measurements of virus load and lymphocyte surface markers were obtained. No correlation between Gag specific CTL frequency and concurrent CD4 cell count was observed. A significant inverse relationship was observed between HIV-1 Gag-specific CTL frequency and provirus load as measured by polymerase chain reaction. Subjects with higher CTL frequencies were found to have more stable CD4 cell counts over time. These results provide additional evidence to support the concept that the predominant role of this virus-specific cellular immune response is to limit viral replication and CD4 cell loss in HIV-1 infection. PMID- 9207358 TI - Early manifestations of disseminated Mycobacterium avium complex disease: a prospective evaluation. AB - A nested case-control study was conducted in two trials of prophylaxis for Mycobacterium avium complex (MAC) infection to describe the specific signs, symptoms, and laboratory abnormalities of MAC disease in AIDS. Patients had < or =200/mm3 CD4 cells and a prior AIDS-defining illness. Of 571 patients, 102 (17.9%) developed MAC bacteremia during a mean follow-up of 256 days. Among cases of MAC disease, 90 were compared with 180 matched controls. Patients with MAC disease were more likely than controls to have lower weights (66.3 vs. 71.1 kg, P = .001) and Karnofsky scores (74.3 vs. 84.4, P < .001); a higher proportion had fever (48% vs. 26%, P = .003), abdominal pain (23% vs. 13%, P =.05), decreased hemoglobin levels (10.9 vs. 12.1 g/dL, P < .001), and elevated alkaline phosphatase (203 vs. 138 U/L, P=.04) and lactate dehydrogenase (334 vs. 280 U/L, P = .02) levels. Characteristics of MAC disease that occurred before bacteremia were weight loss (3 months prior), fever (2 months), and anemia and elevated lactate dehydrogenase (1 month). These data suggest that patients have symptomatic MAC disease for several months prior to the occurrence of bacteremia. PMID- 9207360 TI - Comparison of a tuberculin interferon-gamma assay with the tuberculin skin test in high-risk adults: effect of human immunodeficiency virus infection. AB - A novel, whole blood interferon-gamma (IFN-gamma) assay was evaluated to determine its suitability for detecting Mycobacterium tuberculosis exposure in intravenous drug users with or without human immunodeficiency virus (HIV) infection. Whole heparinized blood was incubated overnight in separate wells with tuberculin purified protein derivative (PPD), saline, and mitogen controls. Levels of IFN-gamma in plasma supernatants were determined by rapid ELISA. Participants were then administered the tuberculin skin test (TST) and tested for cutaneous anergy. The whole blood IFN-gamma test agreed (89%-100%) with a positive TST in both HIV-seropositive and -seronegative subjects, but reactivity to PPD was more detectable by the whole blood assay among those with negative TSTs or anergy. TST induration diameter and IFN-gamma responses were correlated (Spearman's p = .45, P = .0001), but both responses were blunted by HIV infection. In summary, tuberculin reactivity appears to be more detectable by the whole blood IFN-gamma assay than by TST, and the assay requires no return visit for test reading. PMID- 9207359 TI - Surrogate marker of preclinical tuberculosis in human immunodeficiency virus infection: antibodies to an 88-kDa secreted antigen of Mycobacterium tuberculosis. AB - Antibodies to purified, size-fractionated secreted proteins of Mycobacterium tuberculosis in sera from patients with human immunodeficiency virus (HIV) infection and active tuberculosis (HIV/TB patients), and in stored sera obtained from the same patients prior to clinical manifestation of TB, were evaluated by ELISA, and the repertoire of antigens recognized was analyzed by immunoblotting. Compared with non-HIV/TB patients, HIV/TB patients had lower levels of anti mycobacterial antibodies, and these were directed toward a restricted set of antigens. Antibodies to an 88-kDa secreted antigen were present in the sera of 74% of HIV/TB patients during the years (1.5-6) prior to manifestation of active, clinical tuberculosis, although only 66% were positive by the time tuberculosis was diagnosed. The presence of antibodies to the 88-kDa antigen can serve as a surrogate marker for identifying HIV-infected persons with active, subclinical disease who are at a high risk of developing clinical tuberculosis. PMID- 9207361 TI - Sensitization to lipopolysaccharide in mice with asymptomatic viral infection: role of T cell-dependent production of interferon-gamma. AB - The interplay between viral infection and lipopolysaccharide (LPS) was studied. Infection with a noncytopathogenic virus, lymphocytic choriomeningitis virus (LCMV), was found to sensitize mice to low doses of LPS. In vivo, this hypersensitivity correlated with hyperproduction of tumor necrosis factor-alpha (TNF-alpha), and in vitro, LPS-stimulated splenic adherent cells produced increased amounts of TNF-alpha. Hyperproduction of TNF-alpha was temporally correlated with virus-induced production of interferon-gamma (IFN-gamma); only marginally increased IFN-gamma and TNF-alpha production was observed in LCMV infected, T cell-deficient mice and in mice infected with vesicular stomatitis virus, a virus that induces much less T cell activation than does LCMV. Finally, LCMV infection was much less efficient in priming IFN-gamma knockout mice for hyperproduction of TNF-alpha. These findings indicate that clinically silent viral infections may induce hypersensitivity to LPS through T cell activation and subsequent production of IFN-gamma; this sensitizes monocytes/macrophages for hyperproduction of TNF-alpha. PMID- 9207362 TI - Dr fimbriae operon of uropathogenic Escherichia coli mediate microtubule dependent invasion to the HeLa epithelial cell line. AB - Escherichia coli Dr adhesin and decay-accelerating factor (DAF) receptor-mediated interaction was proposed as the mechanism of ascending urinary tract infection (UTI) and chronic interstitial nephritis. This report provides novel evidence for Dr fimbriae operon-mediated invasive capacity of Dr+ E. coli. Insertional mutants draE, draC, and draB, and adherent draD and UV-inactivated BN406 were unable to enter HeLa cells. Complementation of the dra mutation restored invasiveness. Internalization was inhibited by anti-Dr fimbriae IgG (100%), anti-SCR-3 domain of DAF (75%), and nocodazole (95%). Increased receptor-ligand density occurred at the site of internalization. Internalized Dr+ E. coli did not significantly multiply in the HeLa cell line. Accordingly, the dra operon and DAF were required for microtubule-dependent internalization of E. coli to HeLa cells. The relatively low invasion and multiplication rates of Dr+ E. coli may hypothetically contribute to the postattachment steps of ascending UTI and chronic renal infection. PMID- 9207363 TI - Neonatal hypersusceptibility to endotoxin correlates with increased tumor necrosis factor production in mice. AB - Septic shock is a major cause of mortality in neonates. The hypothesis was tested that neonatal age is associated with altered sensitivity to shock-inducing bacterial products or proinflammatory cytokines (or both). Mice of different ages were inoculated with various doses of lipopolysaccharide (LPS), superantigenic staphylococcal enterotoxin B (SEB), or recombinant tumor necrosis factor-alpha (rTNF-alpha), alone or in combination with the sensitizing agent D-galactosamine. Neonatal mice were markedly more susceptible to LPS-induced lethality but more resistant to SEB than were adults (P < .05). Mice of different ages did not differ, however, in their sensitivity to lethal activities of rTNF-alpha. Neonatal susceptibility to LPS and SEB correlated directly with plasma TNF-alpha but not IFN-gamma levels, which was confirmed by TNF-alpha and IFN-gamma blockade experiments. These data document marked age-related differences in the pathophysiology of septic shock and suggest that IFN-gamma is not an obligatory mediator of either LPS- or SEB-induced lethality in neonates. PMID- 9207364 TI - A genetic-based evaluation of the principal tissue reservoir for group A streptococci isolated from normally sterile sites. AB - The primary sites of infection and principal reservoirs for transmission of group A streptococci are the nasopharyngeal mucosa and the impetigo lesion. However, pharyngitis and impetigo are rarely observed prior to invasive disease, and, thus, the origin of invasive strains is largely unknown. As part of an active surveillance program, group A streptococci were obtained from normally sterile tissue sites of Connecticut residents during a 6-month period. Organisms were analyzed for genetic markers that distinguish between strains that use the nasopharynx versus an impetiginous lesion as their primary site for infection. The nasopharyngeal marker was observed for most sterile-site isolates, suggesting that the upper respiratory tract is the principal reservoir from which organisms causing invasive disease are disseminated. Genotypic analyses of sterile-site isolates support the view that additional factors, aside from a recent emergence of a few virulent clones, are important contributors to invasive group A streptococcal disease. PMID- 9207365 TI - Morphine induces sepsis in mice. AB - Gram-negative sepsis and subsequent endotoxic shock remain major health problems in the United States. The present study examined the role of morphine in inducing sepsis. Mice administered morphine by the subcutaneous implantation of a slow release pellet developed colonization of the liver, spleen, and peritoneal cavity with gram-negative and other enteric bacteria. In addition, the mice became hypersusceptible to sublethal endotoxin challenge. The effects were blocked by the simultaneous implantation of a pellet containing the opioid antagonist naltrexone. These findings show that morphine pellet implantation in mice results in the escape of gram-negative organisms from the gastrointestinal tract, leading to the hypothesis that morphine used postoperatively or chronically for analgesia may serve as a cofactor in the precipitation of sepsis and shock. In addition, morphine-induced sepsis may provide a physiologically relevant model of gram negative sepsis and endotoxic shock. PMID- 9207366 TI - Clostridial gas gangrene: evidence that alpha and theta toxins differentially modulate the immune response and induce acute tissue necrosis. AB - The rapid extension of necrosis and an absence of polymorphonuclear leukocytes (PMNL) at the site of infection are two hallmarks of Clostridium perfringens gas gangrene. While both alpha and theta toxins profoundly affect PMNL function and viability in vitro, their roles in muscle destruction and impairment of the inflammatory response in vivo have not been investigated. Comparative histopathologic examinations were performed on animals infected with either wild type C. perfringens, or isogenic, toxin-deficient mutants of C. perfringens. Tissue destruction was modest in animals infected with the alpha toxin-deficient mutant; destruction was more pronounced in tissues infected with the theta toxin deficient mutant or the wild-type strain. alpha and theta toxins also displayed differing abilities to modulate the inflammatory response. Histopathologic studies in which recombinant toxins were injected together with killed, washed C. perfringens further substantiated these tissue-destructive and differential antiinflammatory effects. PMID- 9207367 TI - Helicobacter pylori reinfection is virtually absent after successful eradication. AB - This study examined whether reinfection or recrudescence accounts for the reappearance of Helicobacter pylori infection after apparent successful eradication. In a prospective study, 173 patients cured from H. pylori infection underwent follow-up endoscopies, with biopsies for culture and histopathology, every 3 months during the first year after treatment. Subsequently, elective half yearly endoscopies were performed in 124 patients; the remaining 49 underwent follow-up endoscopy only in 1995. At reappearing infection, DNA profiles of pretreatment and recurrent strains were compared. After 3.5 years (range, 1.0 9.2), H. pylori infection recurred in 9 patients (5.2%). Reappearing infections were classified as endoscopically transmitted reinfection (n = 2), unclassified because of loss of pretreatment isolate (n = 1), or recrudescence (identical DNA patterns before and after treatment; n = 6). The reappearance rate of infection, discarding endoscopic transmission, was 1.2% (7/601 H. pylori-negative patient years). There was virtually no reinfection with H. pylori after eradication in this adult Western population. These data do not rule out acquisition of H. pylori. PMID- 9207369 TI - Experimental infection of young specific pathogen-free cats with Bartonella henselae. AB - Eighteen 12-week-old specific pathogen-free cats, blood culture- and serum antibody-negative for Bartonella henselae, were randomly allocated to groups and were intravenously inoculated with 10(10) (group 1), 10(8) (group 2), or 10(6) (group 3) B. henselae or with saline (group 4) or were not inoculated (group 5). Cats were humanely killed at 2, 4, 8, 16, and 32 weeks after inoculation. All B. henselae-inoculated cats were bacteremic by 2 weeks after infection. Bacteremia persisted until 32 weeks after infection in 1 cat. Cats in groups 1 and 2 had fever (>39.7 degrees C) and partial anorexia by 2 weeks after infection that lasted 2-7 days. All infected cats had Bartonella-specific IgM and IgG serum antibodies and lymphocyte blastogenic responses. Histopathologic lesions were observed in multiple organs of infected cats through 8 weeks after infection. Cats were readily infected with B. henselae by intravenous inoculation, developed histopathologic lesions that apparently resolved, and developed B and T lymphocyte responses to infection. PMID- 9207368 TI - Evaluation of Peru-15, a new live oral vaccine for cholera, in volunteers. AB - A new live oral cholera vaccine, Peru-15, was studied for safety, immunogenicity, and excretion in 2 groups of healthy volunteers. Twelve inpatient volunteers received freshly harvested vaccine in doses of either 10(7) or 10(9) cfu. Subsequently 50 outpatient volunteers received freeze-dried vaccine in doses of 10(8) or 10(9) cfu or placebo in a three-cell, double-masked, placebo-controlled trial. The strain was well tolerated at all dose levels, and it stimulated high levels of vibriocidal antibodies in most inpatient volunteers and in all outpatient volunteers. Although antitoxin responses were less frequent and of lower magnitude than the vibriocidal responses, antitoxin responses were seen in >60% of the outpatient volunteers. About 60% of the volunteers excreted the vaccine in their feces; however, fecal excretion did not correlate with serologic responses. It is concluded that Peru-15 is a safe and immunogenic oral vaccine for cholera. PMID- 9207370 TI - Induction of protective Th1 responses to Candida albicans by antifungal therapy alone or in combination with an interleukin-4 antagonist. AB - Resistance or susceptibility to disseminated and mucosal Candida albicans infections in mice correlates with the development of protective or nonprotective T helper (Th) cell responses. To determine whether immunomodulatory activity on Th cell functions is an effect beyond that provided by antifungal therapy, mice with disseminated or gastrointestinal infection were treated with amphotericin B or fluconazole and assessed for mortality, fungus burden in the organs, and parameters of Th cell-dependent immunity. Both antimycotics produced protective CD4+ Th1 cell responses, as revealed by increased production of interleukin (IL) 12 and interferon-y, decreased production of IL-4, delayed-type hypersensitivity to fungal antigen, and the disappearance of antigen-specific IgE. Concomitant neutralization of endogenous IL-4 greatly increased the antifungal efficacy and the Th1-promoting activity of both agents. These results indicate that successful antifungal therapy alone or in combination with cytokine antagonists may rely on the induction of an appropriate Th antifungal cell response. PMID- 9207371 TI - A randomized, double-blind comparison of itraconazole oral solution and fluconazole tablets in the treatment of esophageal candidiasis. AB - This multicenter, randomized, double-blind study compared the efficacy and safety of itraconazole oral solution and fluconazole tablets in the treatment of esophageal candidiasis. One hundred twenty-six immunocompromised patients with esophageal candidiasis were treated with itraconazole oral solution or fluconazole tablets (both at 100-200 mg) once daily for 3-8 weeks, for 2 weeks beyond the resolution of symptoms, and were then followed for 4 more weeks. Severity of symptoms was assessed weekly during treatment and every 2 weeks during follow-up. Patients treated with itraconazole oral solution had a rate of clinical response (cured or improved) comparable to that of patients treated with fluconazole (94% vs. 91%). The mycologic eradication rate was 92% for itraconazole and 78% for fluconazole. Both treatments were well tolerated. Results from treatment with once-daily itraconazole oral solution was clinically comparable to those with fluconazole and is an alternative for the treatment of esophageal candidiasis in immunocompromised patients. PMID- 9207372 TI - Activation of gammadelta T cells in malaria: interaction of cytokines and a schizont-associated Plasmodium falciparum antigen. AB - A soluble Plasmodium falciparum antigen that specifically stimulates gammadelta T cells has been found associated predominantly with schizonts rather than ring forms, trophozoites, or gametocytes. This schizont-associated antigen (SAA) is resistant to protease digestion, is anionic at pH 8.5, is heat- and pH-resistant, and contains a phosphate group(s) that is crucial for biologic activity. Partially purified SAA induced proliferative responses and interferon-gamma production by gammadelta T cells. These stimulatory effects were greatly enhanced by monocyte-derived cytokines, interleukin (IL)-10, IL-12, and IL-1beta, but not by tumor necrosis factor-alpha. Taken together, these results suggest that concurrent stimulation of gammadelta T cells by SAA and by cytokines released from activated monocytes (IL-10, IL-12, IL-1beta) may represent the major mechanism underlying the selective activation of gammadelta T cells that is consistently observed in clinical cases of P. falciparum infection. PMID- 9207373 TI - Transmission intensity and its relationship to infection and disease due to Wuchereria bancrofti in Papua New Guinea. AB - This study describes the relationship between transmission intensity and infection and disease due to Wuchereria bancrofti in an endemic area of Papua New Guinea. The prevalence of microfilaremia in the entire study population was 66%. Of 1892 persons examined, 6.2% and 12.3% had lymphedema of the legs and hydroceles, respectively. The prevalences of microfilaremia and clinical morbidity were lowest in persons <20 years old and increased progressively with age. Annual transmission potential and annual infective biting were monitored in five villages where Anopheles punctulatus and Anopheles koliensis are the only vectors of W. bancrofti. Both measures of the entomologic inoculation rate were positively associated with the village-specific microfilarial rate, mean intensity of microfilaremia, and prevalence of leg edema. These data indicate that transmission intensity is a major determinant of patent infection and morbidity rates in bancroftian filariasis. PMID- 9207374 TI - Summary: Nagasaki enterohemorrhagic Escherichia coli meeting and workshop. PMID- 9207375 TI - Analysis of the systemic and intrathecal humoral immune response in progressive multifocal leukoencephalopathy. AB - Progressive multifocal leukoencephalopathy (PML) is a subacute viral infection of oligodendrocytes by JC virus occurring almost exclusively in immunocompromised patients. By use of partially purified recombinant VP1 as antigen, the IgG response was analyzed by a quantitative ELISA of paired cerebrospinal fluid (CSF) and serum samples. An intrathecal immune response to VP1, defined as an antibody specificity index of CSF to serum antibody titers > or =1.5, was found in 76% of PML patients (47/62) but in only 3.2% of controls (5/155) (P < .001). Intra-blood brain barrier synthesis of VP1-specific IgG antibodies is 76% sensitive and 96.8% specific for the diagnosis of PML. Furthermore, the excellent correlation (r = .985) between the plasma cell count in brain tissue and the humoral intrathecal immune response to VP1 in PML patients suggests a role for B cells in this disorder. PMID- 9207377 TI - Antisense-mediated resistance to measles virus infection in HeLa cells. AB - Endogenous expression of antisense RNA in transfected cells has been explored for use in blocking cellular gene expression and for its antiviral potential. Antisense strategies were used with the goal of blocking measles virus (MV) infection. A recombinant expression plasmid was designed to produce antisense oligonucleotides targeted to the 5' end of the MV nucleocapsid protein mRNA. This construct was transfected into HeLa cells. The transfected cell line and a control cell line expressing a random RNA comprising the same nucleotides were infected with MV and assessed for viral resistance by observation of cytopathic effect (CPE); infectious virus was quantified by viral plaque assay. Both cell lines were also infected with a related paramyxovirus, mumps virus, as a specificity control. Both CPE and infectious virus were reduced by approximately 90% in the antisense-expressing line compared with that in control cells or transfectant cells expressing random RNA. There was no evidence of resistance to infection with mumps virus in any cell line. PMID- 9207376 TI - Viral interleukin-10 in chronic active Epstein-Barr virus infection. AB - Viral interleukin-10 (IL-10), a product of the Epstein-Barr virus (EBV) replication gene BCRF1, shares extensive structural and functional similarity with the human cytokine IL-10. Both viral and human IL-10 inhibit T cell growth and interferon-gamma production. With two ELISAs, one that recognized both human and viral (total) IL-10 and the other specific for viral IL-10, IL-10 was measured in serum or plasma from 34 patients with chronic active EBV infection (CAEBV) and from 15 healthy controls. Of the patients, 56% had measurable total IL-10 and 29% had measurable viral IL-10. In contrast, total IL-10 was detectable in only 2 of 15 controls and viral IL-10 was undetectable. Thus, many patients with CAEBV have abnormally high levels of circulating IL-10 that may contribute to disease pathogenesis by inhibiting host immunity. PMID- 9207378 TI - Chronic varicella-zoster virus skin lesions in patients with human immunodeficiency virus are related to decreased expression of gE and gB. AB - The pathogenesis of chronic, verrucous varicella-zoster virus (VZV) cutaneous lesions in human immunodeficiency virus (HIV)-infected persons is unknown. It has been hypothesized that these lesions are due to an altered pattern of virus gene expression. Immediate early and late (L) gene expression in five chronic verrucous VZV lesions, four full-blown herpes zoster vesicular lesions in HIV infected persons, and eight vesicular herpes zoster lesions in immunocompetent individuals was semiquantitatively assessed immunohistochemically using specific antibodies to the IE63, gE (L), and gB (L) proteins. All patients had evidence of IE63 expression in keratinocytes; however, gE expression was either weak or absent in keratinocytes of three verrucous lesions, and gB was either weak or absent in two. These results suggest that chronic VZV skin lesions are associated with diminished gE and gB expression. It is inferred that the VZV behavior in keratinocytes may vary from a latency-like state to a fully developed, productive infection. PMID- 9207379 TI - Antagonism between human immunodeficiency virus type 1 protease inhibitors indinavir and saquinavir in vitro. AB - Human immunodeficiency virus type 1 (HIV-1) protease inhibitors are a promising class of antiretroviral agents that compromise enzymatic function through substrate mimicry. The in vitro susceptibility of a panel of HIV-1 clinical isolates demonstrating various drug resistance phenotypes to combinations of the HIV-1 protease inhibitors saquinavir and indinavir was determined. Antiviral effect was assessed by an HIV-1 p24 antigen reduction assay in phytohemagglutinin stimulated peripheral blood mononuclear cells after harvesting of cell-free supernatant fluids at peak antigen production (days 4-7). Drug interactions were determined by median-dose-effect analysis, with the combination index (CI) calculated at several inhibitory concentrations (IC50, IC75, IC90, IC95, IC99). The interactive effects ranged from synergy at low efficacy doses to antagonism at higher doses against a pan-susceptible clinical isolate of HIV-1. Against a zidovudine-resistant isolate as well as a multidrug-resistant isolate, the combination of saquinavir and indinavir demonstrated antagonism at all doses. PMID- 9207380 TI - Clinical relevance of parvovirus B19 as a cause of anemia in patients with human immunodeficiency virus infection. AB - Parvovirus B19 (B19) DNA was detected by dot blot hybridization in sera from 5 (17%) of 30 human immunodeficiency virus (HIV)-infected patients with hematocrits (HCT) of < or =24 and 4 (31%) of 13 HRV-infected patients with HCT of < or =20, suggesting that B19 is a reasonably common cause of severe anemia in HIV infection. The anemia promptly remitted after immunoglobulin therapy in 3 of 4 treated patients. The presence of IgM to B19, the clinical circumstance in which anemia developed, and the marrow morphology were poor predictors of chronic B19 infection. DNA hybridization studies of sera from 191 HIV-infected and 117 HIV seronegative homosexual males attending a clinic in the Seattle area revealed that 1 (0.5%) and 2 (2%) samples, respectively, from the 2 groups contained B19. However, when assayed by polymerase chain reaction (PCR), 5% of the serum samples from HIV-infected persons and 9% from uninfected persons contained B19, although each had an HCT of > or =40. The data argue that anemia results from chronic high titer B19 infection. Although a negative PCR assay excludes this diagnosis, DNA hybridization may be the more specific serum test. PMID- 9207381 TI - Vitamin E supplementation decreases lung virus titers in mice infected with influenza. AB - Effects of vitamin E (E) supplementation on influenza infection were examined in young and old C57BL/6NIA mice fed 30 or 500 ppm of E for 6 weeks and subsequently infected with influenza A/Port Chalmers/1/73 (H3N2). Old mice fed 30 ppm of E had significantly higher lung virus titers on days 2 and 7 after infection than young mice fed 30 ppm of E. Titers on all 3 days were significantly lower in old mice fed 500 ppm of E than in those fed 30 ppm. Significant effects of E on lung virus titers in young mice were observed on only day 5, but E caused more reduction of virus titers in old than in young mice (25-fold vs. 15-fold). An age-associated decline in NK cell activity was restored by 500 ppm of E in old but not young mice. Pulmonary cytotoxic T lymphocyte activity on day 7 was not affected by age or E. These experiments demonstrate that high doses of E significantly enhance influenza viral clearance in aged mice but only modestly affect young mice. PMID- 9207382 TI - Latent human papillomavirus infection in pregnant women at term: a case-control study. AB - Cervicovaginal lavages from 752 pregnant women at term were investigated by polymerase chain reaction to evaluate human papillomavirus (HPV) infection prevalences and were compared with cervicovaginal samples from two series of nonpregnant subjects (504 healthy women attending a family planning service and 560 symptomatic patients attending a vaginitis outpatient service). The odds ratios (ORs) of HPV infection were computed by conditional logistic regression analysis on age-matched sets. In pregnant women, the overall risk of HPV infection was about the same as in nonpregnant healthy subjects (adjusted OR, 0.90; 95% confidence interval [CI], 0.51-1.58) and was 50% less than in patients with symptomatic vaginitis (adjusted OR, 0.48; 95% CI, 0.30-0.76). Moreover, the prevalence of oncogenic HPV types 16 or 18 (or both) was lower in pregnant women (P = .015 and P = .0018 respectively). PMID- 9207383 TI - Genetic evidence for mother-to-infant transmission of hepatitis G virus. AB - Mother-to-infant transmission of hepatitis G virus (HGV [or GBV-C]) was studied in sera from 42 mothers at high risk for bloodborne infections and from their 45 infants (3 twin pairs). Seven (17%) of the mothers had HGV RNA in serum by a polymerase chain reaction assay. One of the 8 (12.5%) infants born to HGV infected mothers became positive for HGV at 3 months of age. He remained HGV infected throughout the study (42 months), with no signs of liver disease. His twin sister remained HGV-negative despite the presence of serum and salivary HGV in both the mother and the brother. Analysis of HGV sequences from the infected mother and the infected child confirmed a genetic link between the virus of the mother and the infected child. Thus, mother-to-infant transmission of HGV, presumably occurring at partus, may cause persistent HGV viremia. PMID- 9207384 TI - Recovery of cytomegalovirus and herpes simplex virus from upper and lower genital tract specimens obtained from women with pelvic inflammatory disease. AB - Lower genital tract specimens and endometrial biopsies from 147 women with pelvic inflammatory disease (PID) and surgical specimens (fallopian tubes, ovaries, or both) from 22 women with PID and 37 women without PID were cultured for cytomegalovirus (CMV) and herpes simplex virus (HSV), as well as for organisms commonly associated with PID. CMV was isolated from 39 cervical or endometrial samples from 30 (20.4%) of 147 women with PID and from ovaries or fallopian tubes from 5 (22.7%) of 22 women with PID, but CMV was not recovered from surgical specimens obtained from 37 women undergoing surgery for tubal ligation, ectopic pregnancy, or other gynecologic conditions (P = .005). HSV was isolated from cervical samples obtained from 5 (3.4%) of 147 women with PID but not from any endometrial or surgical specimens. These data suggest that CMV, but not HSV, may contribute to the pathogenesis of PID in some patients. PMID- 9207385 TI - A self-administered technique for the detection of sexually transmitted diseases in remote communities. AB - The control of sexually transmitted diseases (STDs) in remote rural communities would be enhanced by a sensitive self-administered method for the detection of asymptomatic infection. Results of conventional methods for the detection of STDs were compared with results of tampon-collected specimens analyzed by polymerase chain reaction (PCR) for 480 women. Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis were detected by routine methods in 4 (1%), 14 (3%), and 41 (9%) samples, respectively, while PCR detected these organisms from 52 (11%), 26 (5%), and 75 (16%) tampons, respectively. The detection of each organism was significantly greater by PCR in tampon-collected samples than by routine conventional methods (P < .01). Discrepant results were confirmed by separate primers in 40 of 48 specimens for N. gonorrhoeae, in 11 of 12 specimens for C. trachomatis, and in 31 of 32 specimens for T. vaginalis. Tampons tested by PCR provide an acceptable and sensitive method for detection of STDs in women living in remote areas. PMID- 9207386 TI - Isolation of Chlamydia pneumoniae from a carotid endarterectomy specimen. AB - Chlamydia pneumoniae has been associated with atherosclerotic cardiovascular disease by both seroepidemiologic studies and direct detection of the organism in atherosclerotic plaque by electron microscopy (EM), immunocytochemistry (ICC), and polymerase chain reaction (PCR). Despite the frequent detection of the organism in atheromatous cardiovascular specimens by these methods, only 1 cardiovascular isolate of C. pneumoniae, obtained from a coronary artery, has been previously reported. This study reports the isolation of C. pneumoniae from a prospectively obtained carotid endarterectomy specimen. The organism appears to be identical to other C. pneumoniae isolates by EM morphology, reactivity to species-specific monoclonal antibodies, and Southern hybridization analysis of chromosomal digests using C. pneumoniae-specific DNA probes. C. pneumoniae was detected by PCR or ICC (or both) in 11 (69%) of 16 other endarterectomy specimens tested by both of these methods. These results provide further evidence for an association of C. pneumoniae and atherosclerosis by confirming the presence of viable bacteria within atherosclerotic plaque. PMID- 9207387 TI - Pretreatment with a 55-kDa tumor necrosis factor receptor-immunoglobulin fusion protein attenuates activation of coagulation, but not of fibrinolysis, during lethal bacteremia in baboons. AB - Baboons (Papio anubis) receiving a lethal intravenous infusion with live Escherichia coli were pretreated with either a 55-kDa tumor necrosis factor (TNF) receptor-IgG fusion protein (TNFR55:IgG) (n = 4, 4.6 mg/kg) or placebo (n = 4). Neutralization of TNF activity in TNFR55:IgG-treated animals was associated with a complete prevention of mortality and a strong attenuation of coagulation activation as reflected by the plasma concentrations of thrombin-antithrombin III complexes (P < .05). Activation of fibrinolysis was not influenced by TNFR55:IgG (plasma tissue-type plasminogen activator and plasmin-alpha2-antiplasmin complexes), whereas TNFR55:IgG did inhibit the release of plasminogen activator inhibitor type I (P < .05). Furthermore, TNFR55:IgG inhibited neutrophil degranulation (plasma levels of elastase-alpha1-antitrypsin complexes, P < .05) and modestly reduced release of secretory phospholipase A2. These data suggest that endogenous TNF contributes to activation of coagulation, but not to stimulation of fibrinolysis, during severe bacteremia. PMID- 9207388 TI - Proteases from Aspergillus fumigatus induce release of proinflammatory cytokines and cell detachment in airway epithelial cell lines. AB - Aspergillus fumigatus is a pathogen causing diverse respiratory disorders. Several studies have suggested that fungal proteases may play a role in the pathogenicity of fungi. Since the airways are the most common route for entry of A. fumigatus, this study focused on the ability of fungal proteases to induce the release of proinflammatory cytokines and to cause cell detachment in human pulmonary epithelial cell lines. It was shown that fungal serine protease activity induced the production of interleukin (IL)-8 and IL-6 and monocyte chemotactic protein-1 and caused cell detachment in a dose-dependent fashion. Chymostatin, antipain, phenylmethylsulfonyl fluoride, and heat treatment completely inhibited fungal protease activity, cytokine production and cell detachment; antileukoprotease partially inhibited these activities. By causing cell detachment, fungal proteases may decrease the physical barrier function of the epithelium; however, by eliciting a cytokine response, the epithelium may signal the mucosal inflammatory response against A. fumigatus. PMID- 9207389 TI - Oxamniquine cures Schistosoma mansoni infection in a focus in which cure rates with praziquantel are unusually low. AB - An outbreak of Schistosoma mansoni in northern Senegal was observed in 1988, and chemotherapy with praziquantel in this recently established focus resulted in very low parasitologic cure rates. Among other explanations, the emergence of a praziquantel-tolerant parasite strain was feared. To study this hypothesis further, 138 persons with endemic S. mansoni infection were randomly allocated to treatment with either 20 mg/kg oxamniquine or 40 mg/kg praziquantel. Parasitologic cure rates at 6 weeks were significantly higher in the oxamniquine group (79%) compared with those in the praziquantel group (36%; P = .0043). The reduction in egg counts was generally good, but 12% less reduced in the praziquantel group. These results confirm that cure rates with praziquantel were abnormally low, whereas oxamniquine performed satisfactorily, as in other areas in which S. mansoni is endemic. The possibility of a praziquantel-tolerant S. mansoni strain must therefore be studied carefully. PMID- 9207391 TI - What is the nature of the hairy cell and why should we be interested? PMID- 9207390 TI - Human T cell lymphotropic virus type II and human immunodeficiency virus type 1 disease progression. PMID- 9207392 TI - HHV-8-positive body-cavity-based lymphoma: a novel lymphoma entity. PMID- 9207393 TI - The human thrombin receptor and proteinase activated receptor-2 genes are tightly linked on chromosome 5q13. AB - The thrombin receptor (TR) and proteinase activated receptor-2 (PAR-2) may represent the prototypes of an emerging family of cell-surface receptors that effect cell activation events mediated by serine proteases generated during inflammatory, fibrinolytic or haemostatic-regulated pathways. To further characterize the molecular genetics of these receptors, we have refined the genetic and physical mapping of both PAR-2 and TR. Utilization of two distinct radiation hybrid mapping panels with different levels of resolution demonstrated that both genes are tightly linked to the microsatellite markers D5S424, D5S1977, D5S2529 and D5S2596 (in order of decreasing LOD scores, from 13.7 for D5S424 to 7.7 for D5S2596). Physical mapping using yeast artificial chromosomes (YACs) and inversion field gel electrophoresis demonstrated that they are maximally separate by 90 kb. If the association of TR and PAR-2 genes resulted from a relatively recent gene duplication event from a common ancestral gene, these observations provide a general framework for the identification of gene transcripts representing alternative proteolytically activated receptors which may be clustered within this region of the human genome. These observations are especially relevant given recent evidence that murine and human platelets express alternative signalling mechanisms or receptors for thrombin. PMID- 9207394 TI - Influence of thrombopoietin on platelet activation in myeloproliferative disorders. AB - Although thrombopoietin itself does not influence platelet aggregation, it enhances platelet activation in response to certain agonists. We evaluated the effects of thrombopoietin on platelet activation using platelet-rich plasma from 16 patients with myeloproliferative disorders (MPD group) and 16 healthy volunteers (control group). Preincubation with thrombopoietin significantly enhanced platelet aggregation stimulated by ADP, collagen, or epinephrine in the MPD group as well as the control group. However, aggregation induced by 3 micro ADP or 16 microM epinephrine showed significantly less augmentation by thrombopoietin in the MPD group than in the control group. Thrombopoietin significantly shortened the lag time between the addition of 3 microM ADP or 16 microM epinephrine and initiation of secondary aggregation and the lag time between addition of 2 microg/ml collagen and initiation of aggregation in both groups. When platelet-rich plasma was used without adjustment of the platelet count, thrombopoietin itself induced aggregation in two patients. Hypoaggregation after addition of 0.5 microg/ml collagen was observed in seven out of nine patients with normal thrombopoietin levels and only one of six patients with high levels (P = 0.04). Enhancement of 0.5 microg/ml collagen-induced aggregation by thrombopoietin was seen in five out of nine patients with normal thrombopoietin levels and none of the six patients with elevated levels (P = 0.04). These results indicate that platelet activation by certain agonists is enhanced by thrombopoietin in patients with these diseases as well as in normal controls and that the serum thrombopoietin level may regulate the function of circulating platelets in vivo. PMID- 9207395 TI - Measurement of endogenous plasma thrombopoietin in patients with acquired aplastic anaemia by a sensitive enzyme-linked immunosorbent assay. AB - We measured the endogenous plasma concentration of thrombopoietin (TPO) in 76 patients with acquired aplastic anaemia (AA) by a sensitive sandwich enzyme linked immunosorbent assay (ELISA). The plasma TPO concentrations were significantly higher in AA patients when compared to healthy control subjects (P < 0.0001) and there was a significant negative correlation between plasma TPO concentrations and platelet counts in 54 AA patients who had not received any platelet transfusions prior to sampling. On the other hand, there was no statistically significant correlation between the TPO concentrations and platelet counts in 22 AA patients who had previously received platelet transfusions. We studied serial changes of plasma TPO concentration in 24 patients who showed an increase in their platelet counts following bone marrow transplantation or immunosuppressive (IS) therapy. Although a decrease in plasma TPO concentration was observed, levels remained above the range of normal healthy controls even in the patients who attained normal platelet counts following therapy. A decrease in TPO concentrations was observed in only half of the responders to IS therapy. Whether exogenous TPO will result in increased platelet counts in AA patients with high TPO levels remains to be determined. PMID- 9207396 TI - Combination chemotherapy with CHOP for recurrent thrombotic thrombocytopenic purpura. AB - We report the case of a 42-year-old woman with chronic recurrent thrombotic thrombocytopenic purpura. Therapy with corticosteroids, high-dose immunoglobulins, plasma exchange and cyclophosphamide only induced short-lasting remissions during a course of almost 3 months. Following a severe relapse on day 90 after the start of symptoms, polychemotherapy with cyclophosphamide. adriamycin, vincristine and prednisolone (CHOP) was begun. After two cycles of CHOP the patient has stayed in complete remission, with normal platelet counts for more than 9 months to date. PMID- 9207397 TI - The site of destruction of autologous 111In-labelled platelets and the efficiency of splenectomy in children and adults with idiopathic thrombocytopenic purpura: a study of 578 patients with 268 splenectomies. AB - The indication for splenectomy in chronic idiopathic thrombocytopenic purpura (ITP) remains a controversial subject. The mortality rate of persistent thrombocytopenia is very low, except in severe cases. Conversely, the risks of splenectomy are significant (in the present series, morbidity: 4.1% mortality: 1.4%), with a success rate of only 60-75%. It is therefore useful to define a parameter able to predict the efficacy or failure of splenectomy. An analysis of 578 cases of chronic ITP, where the site of platelet destruction has been determined, is presented. 268 of these cases had been splenectomized. When platelet destruction was splenic, 96% of subjects aged 5-30 years and 91% of cases over the age of 30 years obtained a remission. Conversely, when platelet destruction was hepatic or diffuse, failure or incomplete results were observed in 92% of cases. The site of platelet destruction therefore constitutes a parameter which can help the clinician to make the decision to perform splenectomy. PMID- 9207398 TI - Activated protein C resistance can be associated with recurrent fetal loss. AB - As thrombosis of placental vessels may result in recurrent fetal loss, we analysed 39 consecutive women with recurrent fetal loss of unknown cause for activated protein C resistance. Factor V Leiden (FVL) mutation (19 cases) or APC resistance without FVL (nine cases) were found among these 39 women. Evaluation of 128 pregnancies in 19 patients with factor V Leiden mutation and 56 gestations in nine women with acquired APC resistance, revealed over 50% first-trimester abortions and 17% late abortions. Intra-uterine fetal death occurred in nine out of 19 FVL patients (47%). Only 34 of 184 gestations (18%) in hereditary or acquired APC-resistance women resulted in a live birth, with 11 of the 34 (32%) being premature deliveries. These data suggest that, in some patients with recurrent fetal loss, hereditary and acquired APC resistance are potential causes of vascular placental insufficiency. PMID- 9207399 TI - A frequent mutation in the protein S gene results in cryptic splicing. AB - Protein S is a vitamin K dependent coagulation inhibitor. One of several defects in the protein S gene (PROS1) associated with hereditary deficiency is a G --> A transition at position 5 of the splice donor in intron J. Although the mutation has been reported to cause allelic exclusion, we demonstrated low amounts of alternatively spliced ectopic PROS1 transcripts in carriers of this mutation. Sequencing of mutant RNA indicated the use of a cryptic splice site upstream of the common splice donor. The use of the cryptic splice site results in the deletion of 32 nucleotides at the 3' end of exon 10. The new reading frame contains several premature termination signals. PMID- 9207400 TI - Feasibility of prenatal diagnosis and carrier detection in South African haemophilia A patients. AB - The feasibility of DNA diagnosis for haemophilia A was tested in South African patients and families by screening for the common inversion mutation in the factor VIII gene and for the intragenic microsatellite markers in introns 13 and 22. The allele frequencies at the two microsatellite loci were significantly different, with informativity being higher in the Negroid (100%) than the Caucasoid group (67%). In severely affected haemophiliacs the inversion was found in 43% (6/14) of Negroids but in only 32% (13/41) of Caucasoids. Presence of a second common unidentified mutation may account for the low frequency in the latter. Haplotype analysis shows a disproportionately high frequency of an (AC)20 intron 13-(AC)26 intron 22 inversion negative Caucasoid haemophilia chromosome, supporting a founder effect. PMID- 9207401 TI - Factors required for bone marrow stromal fibroblast colony formation in vitro. AB - Marrow stromal fibroblasts (MSFs) are essential for the formation of the haemopoietic microenvironment and bone; however, regulation of MSF proliferation is poorly understood. MSF colony formation was studied in primary mouse and human marrow cell cultures. After a brief exposure to serum, MSF colony formation occurred in the absence of both serum and non-adherent marrow cells, if medium conditioned by marrow cells was present (serum-free conditioned medium, SF-CM). In mouse and human cultures stimulated to proliferate by SF-CM, neutralizing antibodies against PDGF, TGF-beta, bFGF and EGF specifically suppressed MSF colony formation. The degree of supression was species-dependent, with the most profound inhibition achieved in mouse cultures by anti-PDGF, anti-bFGF and anti EGF, and in human cultures by anti-PDGF and anti-TGF-beta. Serum-free medium not conditioned by marrow cells (SFM) did not support MSF colony formation. In mouse cultures in SFM, human recombinant bFGF and bovine natural bFGF were able to partially substitute for the stimulating effect of SF-CM. Other growth factors, including TGF-beta1, TGF-beta2, PDGF, EGF, IL-6, IGF-I and IGF-II, showed no activity when tested alone. In human cultures in SFM, none of the growth factors, alone or in combination, stimulated MSF colony formation. Mouse and human MSFs grown in SF-CM formed bone and a haemopoietic microenvironment when transplantated into immunodeficient mice in vivo, and therefore were functionally equivalent to MSFs generated in the presence of serum. These data indicate that stimulation of the initial proliferation of an MSF precursor cell is complex, and requires participation of at least four growth factors: PDGF, bFGF, TGF-beta and EGF. In addition, mouse and human MSF precursor cells have different requirements for each of the growth factors. PMID- 9207402 TI - The engagement of CD4 surface antigen in the HEL haemopoietic cell line up regulates the transforming growth factor-beta1 (TGF-beta1) promoter activity. AB - In order to investigate the effect of CD4 engagement on the transforming growth factor beta1 (TGF-beta1) promoter activity in haemopoietic progenitors, HEL cells were transiently transfected with a plasmid vector containing -453/+11 nucleotides of the TGF-beta1 promoter fused with the bacterial chloramphenicol acetyltransferase (CAT) gene and then treated with various agonists. Both cross linked CD4mAb and envelope gp120 were able to significantly up-regulate CAT activity with respect to the levels of activation observed in HEL cells treated with cross-linked CD8 mAb or p24. By using deletion mutants of the TGFbeta1 promoter, we found that the minimal DNA sequence still responsive to cross-linked CD4 mAb or gp120 was located between nucleotides -453/-323 of the TGF-beta1 promoter, which contains two activating protein 1 (AP1) binding sites. In electromobility shift assays (EMSA) we could demonstrate that CD4 engagement of HEL cells induced a significant increase of AP1 binding activity at the nuclear level. Furthermore, the steady-state mRNA of endogenous TGF-beta1 showed a small but reproducible increase when HEL cells were treated with cross-linked CD4mAb or gp120. Altogether, these findings suggest that the engagement of CD4 in HEL cells modulates TGF-beta1 expression, acting predominantly at the transcriptional level. PMID- 9207403 TI - Beta-spectrin Campinas: a novel shortened beta-chain variant associated with skipping of exon 30 and hereditary elliptocytosis. AB - Beta-Spectrin Campinas is a novel spectrin variant associated with a shortened beta-chain in a kindred with hereditary elliptocytosis (HE). The propositus and her mother exhibited increased amounts of spectrin dimers and an increase in the alphaI 74 kD fragment from the alpha-chain after partial tryptic digestion of spectrin. The shortened beta-chain appeared as an additional band of approximately 200 kD on SDS-PAGE. In order to delineate the molecular defect of this abnormality at the gene level, reticulocyte mRNA was transcribed into cDNA and the last four exons of the beta-spectrin gene were amplified. Agarose gel of the amplification product of the propositus revealed the expected band of 487 bp as well as a shortened band of approximately 300 bp (size determined on gel). This shortened cDNA amplification product was cloned and nucleotide sequencing revealed the absence of the entire exon 30. In order to determine the underlying mutation responsible for this abnormal splicing, a genomic DNA fragment containing exons 30 and 31 was amplified and nucleotide sequencing revealed a G- >A substitution at the 5' donor splice site consensus sequence of intron 30 (nt + 1 IVS30). The skip splicing observed in this study results in a frameshift, creating a new stop codon and causing a deletion of 129 amino acids at the very COOH-terminus of the protein, thus impairing spectrin dimers self-association. We classified this HE as spherocytic HE because the propositus presented a few spherocytes in addition to many elliptocytes in the blood smear, whereas her mother, who was splenectomized, showed many schizocytes, poikilocytes and spherocytes. PMID- 9207404 TI - Treatment with recombinant human erythropoietin (rHuEpo) in a patient with paroxysmal nocturnal haemoglobinuria: evaluation of membrane proteins CD55 and CD59 with cytofluorometric assay. AB - We describe a 28-year-old man with paroxysmal nocturnal haemoglobinuria (PNH) and a high transfusion requirement. Prior to and during therapy with recombinant human erythropoietin (rHuEpo), we evaluated the levels of 'decay-accelerating factor', CD55, and 'membrane-inhibitor-of-reactive-lysis', CD59, as markers of the disease, whilst CD58, a marker present on leucocytes, was utilized to monitor normal haemopoietic activity. The patient became transfusion independent 1 month after beginning rHuEpo and remains well. The analysis of CD55, CD59 and CD58 suggests that the efficacy of rHuEpo was due to a selective rHuEpo action on normal erythroid clones. PMID- 9207405 TI - Multiple bcl-2/Ig gene rearrangements in persistent polyclonal B-cell lymphocytosis. AB - Persistent polyclonal B-cell lymphocytosis is a benign lymphoproliferative disorder of unknown aetiology occurring exclusively in women, characterized by typical binucleated lymphocytes, polyclonal expansion of B cells and elevated serum IgM. Owing to the role of Bcl-2 oncogene in inhibition of apoptosis, we have investigated the presence of the bcl-2/Ig gene rearrangement. Bcl-2/Ig gene rearrangement was determined by polymerase chain reaction targeting the usual breakpoint regions of the t(14;18). Bcl-2/Ig gene rearrangement was identified in all six patients and, more importantly, multiple rearrangements were present in five patients. The frequency of the bcl-2/Ig gene rearrangement is estimated to be of one translocation in 1 x 10(2) to 1 x 10(3) peripheral blood mononuclear cells. We conclude that persistent polyclonal B-cell lymphocytosis is associated with bcl-2/Ig gene rearrangement. These findings are of clinical importance because these patients may be misdiagnosed as having a leukaemic expression of non-Hodgkin's lymphoma. PMID- 9207406 TI - Prognostic factors in elderly acute lymphoblastic leukaemia. AB - A retrospective study was performed on 46 unselected acute lymphoblastic leukaemia (ALL) elderly patients aged 60 years or more. Only 50% of these patients were included in the EORTC cooperative clinical trials, thus confirming the important selection bias in most of the published series on elderly ALL patients. 43% of the elderly patients achieved a complete remission (CR). The median survival was 10 months and the 5-year overall survival was only 7.6 +/- 4%. In multivariate analysis, W.H.O. performance status and peripheral blast counts at day 7 were found to significantly influence achievement of CR and survival. In patients with W.H.O. performance status > or = 2, 35% died during induction treatment versus 4% in patients with W.H.O. performance status < 2. Patients > 70 years old showed a marked drop of the CR rate (27%) compared to those aged 60-69 (67%), and a very high death rate during the induction period (38% versus 4%). This suggests that ALL protocol treatments should be proposed until 70 years in patients with good-performance status, whereas less intensive treatment should be offered to elderly patients with performance status > or = 2 and/or age > or = 70. Peripheral blast counts at day 7 may help to adjust the treatment during induction phase. PMID- 9207407 TI - Clearance of marrow infiltration after 1 week of therapy for childhood lymphoblastic leukaemia: clinical importance and the effect of daunorubicin. The Medical Research Council's Working Party on Childhood Leukaemia. AB - At the commencement of UKALL XI, a national MRC trial for childhood lymphoblastic leukaemia (ALL), the therapy included a bolus of daunorubicin (DR) on the first 2 d of the protocol. This component of treatment was subsequently withdrawn because of concern about long-term cardiotoxicity. All children both before and after this change of policy had their marrow status at the end of the first week assessed by central review as part of the trial to examine the clinical importance of the rate of disease clearance. This also afforded an opportunity to observe the effect of DR on gross residual disease at an early stage of therapy. 1419 children were studied: 342 received DR ('recipients'), 1077 did not. 44% of the recipients completely cleared their marrow of blast cells after 8 d compared with 13% of the non-recipients (chi2 = 158.2, P < 0.0001). The difference in the proportion with massive residual disease (>80% blasts) was less impressive but there was still a difference in favour of DR recipients (DR 9%, no DR 15%; chi2 = 7.7, P = 0.006). The rate of disease clearance correlated with disease-free survival for both recipients and non-recipients, but there was no significant difference in outcome when comparing the two groups with each other, either in terms of disease-free or relapse-free survival. DR accelerated the rate of blast cell disappearance from the marrow but the difference this made to disease free survival is small or non-existent. It appears to be the relative speed of response to a given therapeutic regimen that is prognostically important rather than the absolute rate of response when comparing one treatment with another. PMID- 9207408 TI - Minimal residual disease with TEL-AML1 fusion transcript in childhood acute lymphoblastic leukaemia with t(12;21). AB - We analysed the TEL-AML1 transcript using reverse transcription-polymerase chain reaction (RT-PCR) in order to detect minimal residual disease (MRD) in seven children with t(12;21)-associated B-lineage ALL. Leukaemic cells with the TEL AML1 transcript appear to be very sensitive to chemotherapy, and may be eradicated in most patients if adequate chemotherapy is given. However, a small number of patients with t(12;21) ALL may relapse under the currently used chemotherapy, and we believe that RT-PCR for detecting MRD with the transcript is a suitable tool for monitoring the efficacy of chemotherapy or impending relapse in these patients. We analysed the TEL-AML1 transcript using reverse transcription-polymerase chain reaction (RT-PCR) in order to detect minimal residual disease (MRD) in seven children with t(12;21)-associated B-lineage ALL. Two sets of primers, TEL exon 5 and AML1 exon 3 or 4, detected two types of transcript in four patients and two other types in two other patients. The two different translocation breakpoints in the AML1 gene with or without splicing out of AML1 exon 3 seemed to result in these four types of transcript in leukaemia samples. PMID- 9207409 TI - Heterogenous response of B lymphocytes to transforming growth factor-beta in B cell chronic lymphocytic leukaemia: correlation with the expression of TGF-beta receptors. AB - We investigated the potential role of transforming growth factor-beta (TGF-beta) on spontaneous and cytokine-induced proliferation of B-cell chronic lymphocytic leukaemia (B-CLL) cells in vitro. Purified B lymphocytes from 21 B-CLL patients were cultured for 5 d in the presence of medium alone, IL-2 and/or IL-10, in the presence or absence of TGF-beta, and proliferation was measured by 3H-thymidine incorporation. TGF-beta inhibited B-cell proliferation in the majority of patients (15/21) but no inhibition was detected in 6/21 patients whatever the type of stimulant used. Addition of neutralizing antibodies to TGF-beta increased spontaneous and cytokine-induced proliferation; this effect was dose dependent and specific because addition of an irrelevant chicken IgG had no effect on B-CLL proliferation. In resistant patients, neutralizing antibodies to TGF-beta did not increase the proliferation. The expression of TGF-beta receptors on B-CLL cells was significantly lower than the one observed on normal CD5+ B lymphocytes for which the sensitivity to TGF-beta inhibition was more marked than in CLL. In addition, we found a strong correlation between the response of leukaemic B cells to TGF-beta inhibitory action and the expression of TGF-beta receptors on these cells. In summary, TGF-beta appears to function in CLL as a negative regulator of B lymphocytes but loss of responsiveness to this factor accompanied by a decrease of TGF-beta receptor expression, might provide a selective advantage to B-CLL lymphocytes. PMID- 9207410 TI - Clonal chromosomal abnormalities as direct evidence for clonality in nasal T/natural killer cell lymphomas. AB - Nasal T/natural killer (NK) cell lymphoma is a distinct clinicopathologic entity which is more prevalent in Asia than in America and Europe. The clonal nature of the infiltrating lymphoid cells is difficult to demonstrate because of the lack of immunologic markers for clonality and the absence of clonal T-cell receptor gene rearrangement in most cases. In this study, clonal chromosomal abnormalities were detected in the tumour cells from four patients with nasal T/NK cell lymphoma. This finding provided direct evidence for clonality of the disease. Moreover, nonrandom cytogenetic abnormalities, including isochromosome for the short arm (p) of chromosome 6, isochromosome for the long arm (q) of chromosome 1, partial deletion of 6q, and aberrations at 11q, were disclosed. Isochromosome 6p was the sole structural abnormality in one patient, which may be a pathognomonic change in nasal lymphoma. PMID- 9207411 TI - Common clonal origin of lymphocytes and plasma cells in splenic lymphoma with villous lymphocytes. AB - In two-thirds of patients with splenic lymphoma with villous lymphocytes (SLVL) a small amount of M-protein can be detected in association with the presence of plasma cells in the peripheral blood (PB) and/or bone marrow (BM). However, it is not known whether lymphoma cells and plasma cells originate from the same clone. In this report we describe a case of SLVL which was characterized by the presence of marked monoclonal gammopathy (IgG-kappa 90 g/l) and increased plasma cells in the BM. In an attempt to elucidate the origin of lymphoma cells and plasma cells, we performed morphological, cytogenetic and molecular studies on PB mononuclear cells (PBMNC) without plasma cells and BMMNC containing 10% plasma cells from this patient. Immunofluorescence showed that lymphoma cells and plasma cells were positive for cytoplasmic gamma heavy and kappa light chains. Well-developed endoplasmic reticulum was observed in the cytoplasmic organelles of PBMNC using an electron microscope. The mean IgG concentration in the 3 d supernatant cultures of PBMNC was 374 +/- 24 microg/l. More than 50% PBMNC differentiated into plasmacytoid cells in 6 d of liquid culture with IL-3 and IL-6. Analysis by two-colour FISH revealed that karyotypic abnormalities of monosomy X and trisomy 17 existed simultaneously in both lymphoma cells and plasma cells. JH gene rearranged bands from PBMNC and BMMNC by Southern blot hybridization were identical, whereas DNAs from PBMNC failed to hybridize with the Cmu probe. These observations strongly suggest that lymphoma cells and plasma cells originate from the same clone, and that plasma cells, as well as lymphoma cells, which have undergone class switch recombination, could produce IgG type M-protein in this case. PMID- 9207412 TI - Identification of 'short-lived' and 'long-lived' patients at presentation of idiopathic myelofibrosis. AB - To contribute to a better knowledge of the prognosis of idiopathic myelofibrosis (IM), the prognostic value of the presenting features in 106 patients diagnosed with IM at a single institution during a 21-year period was retrospectively analysed. Median survival was 59.4 months (95% CI 40.7-75.4). Using univariate analysis, age > 64 years, constitutional symptoms (fever, night sweats, weight loss), Hb < 10 g/dl, circulating blasts (> or= 1%), and serum LDH > 3 times upper normal level were associated with a significantly shorter survival; male sex, platelet count < 100 x 10(9)/l, blood percentage of immature granulocytes (excluding blasts), low cholesterol levels and advanced marrow histological stage had borderline significance. Using multivariate study, only age > 64 years, constitutional symptoms, Hb < 10 g/dl, and circulating blasts retained their prognostic relevance. The latter three variables confirmed their predictive value in patients above and below the series median age, and were able to identify two groups of patients: a low-risk group of 67 patients with none or one bad prognostic factor, in whom IM had an indolent course (median survival 98.8 months, 95% CI 68.7-127.6), and a high-risk group, including 39 patients with two or three factors, with a more aggressive disease (median survival 20.6 months, 95% CI 10-28.2). Finally, the application of two recently proposed scoring systems (in which three prognostic groups are considered) was unable to separate intermediate- from high-risk patients. PMID- 9207414 TI - Prognosis in monoclonal gammopathy of undetermined significance. AB - Eighty-seven patients with monoclonal gammopathy of undetermined significance (MGUS) were followed for a period of 1-20 years, median 91 months. Transformation to multiple myeloma occurred in 14 patients of whom seven died as a consequence of the disease. There were 13 unrelated deaths. The actuarial probability of survival was 80% at 10 years and 44% at 15 years and the probabilities of malignant conversion for the same periods were 17% and 30% respectively. The most significant factor influencing the probability of malignant conversion was the increase of monoclonal protein above the level of 30 g/l during the observation period (P<0.001), followed by an increase of M-protein to more than 50% above the baseline level (P=0.02) and a decreased level of uninvolved immunoglobulins (P=0.054). PMID- 9207413 TI - Biochemical, histomorphometric and densitometric changes in patients with multiple myeloma: effects of glucocorticoid therapy and disease activity. AB - It is unknown whether bone changes which can occur in multiple myeloma (MM) are due to cytokine-induced osteoclastic bone resorption from a clone of abnormal plasma cells or high-dose glucocorticoid therapy. We studied 25 MM patients treated for 1-12 years with combination chemotherapy, subdivided into two groups. Group 1 consisted of 12 patients with stage I and II myeloma and group 2 consisted of 13 patients with stage III MM. Their serum biochemistry, tetracycline-labelled bone histomorphometry and bone densitometry were compared to age- and sex-matched controls. Patients with MM demonstrated increased indices of bone resorption (P < 0.001 versus controls) and, to a lesser extent, increased indices of bone formation (P < 0.01 versus controls). No patient had evidence of a mineralization defect. Lumbar spine, femoral neck and total body bone mineral density measurements (BMD) were significantly lower in group 2 compared with group 1 (P < 0.05). Following 12 months of therapy, lumbar spine BMD decreased by 6.6% (95% CI, 2.7% to -9.3%) and femoral neck BMD decreased by 9.5% (95% CI, 3.2% to -15.9%). In a stepwise regression analysis, cumulative prednisolone dosage (B Coef. = -0.39; P = 0.03) and plasma cell infiltrate (B Coef. = -0.08; P = 0.05) were the most important predictors of lumbar spine bone loss, whereas serum paraprotein (B Coef.= -0.35; P = 0.02) and plasma cell infiltrate (B Coef. = -0.20; P = 0.04) were the most important predictors of femoral neck bone loss. We conclude that MM is characterized by high bone turnover with osteoblast osteoclast uncoupling. Both disease activity and high-dose glucocorticoid therapy may be responsible for the ongoing bone loss seen with MM. PMID- 9207415 TI - CD40 and CD95 induce programmed cell death in the human myeloma cell line XG2. AB - We have previously demonstrated that CD40 stimulation induced a cellular growth arrest of the highly CD40-positive myeloma cell line XG2. To further characterize this inhibition of proliferation, we looked for a possible induction of apoptosis. Since no DNA fragmentation could be detected, we used newly described techniques that enable detection of apoptosis independently of DNA degradation, i.e. supravital exposure to propidium iodide (PI) and Annexin V labelling. We demonstrated that CD40 effectively induced programmed cell death. Furthermore, we have shown that CD95 (Fas) stimulation significantly enhanced the CD40-induced apoptosis. PMID- 9207416 TI - Myelomonoblastic leukaemia cells carrying the PEBP2beta/MYH11 fusion gene are CD34, c-KIT+ immature cells. AB - To clarify the aspects affected by the PEBP2beta/MYH11 fusion gene involved in the inv(16), we analysed immunophenotypes in myelomonoblastic leukaemias. We found high expressions of CD34 and c-KIT antigens in myelomonoblastic cells from all patients carrying this fusion gene, including two with M4 and one CML blastic phase, in contrast to those with M4 without the fusion gene. These findings indicate that immunophenotyping is useful for detecting a leukaemia with the fusion gene in myelomonoblastic leukaemias and that the PEBP2beta/MYH11 gene is involved in immature cells expressing CD34 and c-KIT antigens. PMID- 9207417 TI - Tobacco smoking and risk of haematological malignancies in adults: a case-control study. AB - A retrospective study was conducted in 1216 cases to investigate the possible association between tobacco smoking and the risk of haematological malignancies. A small, but not significant, increase in malignancy was observed in smokers. Significant association was demonstrated between tobacco smoking and acute nonlymphoblastic leukaemia, and myelodysplastic syndromes. The duration and amount smoked increased the risk; heavy smokers presented significant positive associations with overall malignancies, acute nonlymphoblastic leukaemia, myelodysplastic syndromes, and monoclonal gammopathy of undetermined significance, whereas light smokers did not present any significant association. These data support a causal relationship between certain haematological malignancies and tobacco smoking. Further research is needed to examine the risk according to dose-response effect, and the variation in risk according to the histological subtype of the malignancy. PMID- 9207418 TI - Successful treatment of cerebral aspergillosis with a novel triazole (voriconazole) in a patient with acute leukaemia. AB - Invasive aspergillosis is an increasing problem in patients with acute leukaemia, bone marrow transplantation, immunosuppression after solid organ transplantation, or acquired immunodeficiency syndrome. Despite available antifungal treatment, the mortality approaches 100% in patients with dissemination of the infection into the central nervous system (CNS). Using a novel triazole, voriconazole, we successfully treated an Aspergillus brain abscess in a patient with acute leukaemia. Drug levels above the minimal fungicidal concentration for Aspergillus species were detected in cerebrospinal fluid (CSF) specimens, and the treatment achieved an objective response. PMID- 9207419 TI - Angina pectoris occurring during granulocyte colony-stimulating factor-combined preparatory regimen for autologous peripheral blood stem cell transplantation in a patient with acute myelogenous leukaemia. AB - We describe a patient with acute myelogenous leukaemia who developed angina pectoris during pretransplant conditioning for autologous peripheral blood stem cell transplantation (PBSCT); the conditioning regimen consisted of cytotoxic drugs in combination with granulocyte colony-stimulating factor (G-CSF). Neutrophilia and hypercoagulability were observed at the time of angina pectoris. Recurrence of angina pectoris was not seen after nitrate and aspirin therapy. Exercise stress testing performed after PBSCT suggested the presence of myocardial ischaemia. Therefore cases at risk of vascular events should be carefully managed with prophylactic treatment during G-CSF administration. PMID- 9207420 TI - In vivo 'purging' of residual disease in CLL with Campath-1H. AB - We assessed the role of human CD52 antibody (Campath-1H) in six patients with chronic lymphocytic leukaemia (CLL) treated to maximal response with purine analogues (fludarabine/deoxycoformycin) in whom persistent leukaemic infiltration of blood and bone marrow had precluded autologous stem cell transplantation. Five patients achieved haematological and histological complete remission following Campath-1H and one had minimal focal residual CLL in a trephine biopsy. Autologous transplantation was performed in two patients without complications and with rapid haemopoietic engraftment. Treatment with Campath-1H may be of value in eradicating residual disease in CLL and may facilitate high-dose therapy in young patients. PMID- 9207421 TI - Thymic function in adults: evidence derived from immune recovery patterns following myeloablative chemotherapy and stem cell infusion. AB - We studied 45 patients aged 14-66 years who had undergone stem cell transplantation for a variety of malignant conditions at least 12 months previously. Compared to normal controls, they had significantly reduced absolute numbers of CD4+, CD4+ CD45RA+ and CD4+ CD45RO+ T cells and a reduced CD4+ CD45RA+:CD4+ CD45RO+ ratio. In all subsets T-cell numbers were significantly greater 24 months, compared to 12-24 months, after transplantation and there was a nonsignificant trend towards lower T-cell numbers with increasing age. We conclude that the thymus, or putative thymic-equivalent tissue, remains functional in older adults. PMID- 9207422 TI - Pilot study of HLA alloimmunization after transfusion with pre-storage leucodepleted blood products in aplastic anaemia. AB - We have performed a pilot study to examine the incidence of alloimmunization using pre-storage leucocyte-depleted blood products (PLDP) in 16 previously transfused aplastic anaemia (AA) patients with no detectable HLA antibodies. A further eight AA patients with HLA antibodies received HLA-matched PLDP. Leucodepleted apheresed platelets were obtained using either Cobe spectra or Haemonetics system with an integral pall filter. Pall BPF4 filters were used for red cell preparation. Patients' sera were tested for HLA antibodies using lymphocytotoxicity (LCT). Patients who were HLA antibody negative by LCT at study entry were further tested with enzyme-linked immunoassay (ELISA). Out of 16 patients, two (12%) formed anti-HLA antibodies with a median follow-up of 9 months (range 1-15), but did not display platelet refractoriness to random donor platelets. Two patients were inadvertently transfused with non-leucodepleted blood products when later referred back to their local hospital. Both subsequently demonstrated HLA antibodies by LCT and became platelet refractory. These results contrast with a 50% incidence of HLA alloimmunization in a control group of AA patients transfused prior to this study with non-PLDP. HLA antibodies could no longer be detected by LCT in follow-up of three out of eight patients with HLA antibodies at study entry. Only one patient experienced non-haemolytic febrile transfusion reactions (NHFTR). We conclude that PLDP reduce the risk of alloimmunization even in previously transfused AA patients, PLDP are associated with a low incidence of NHFTR, and all new AA patients should receive PLDP from diagnosis. PMID- 9207423 TI - All-trans retinoic acid induced thrombocytosis in a patient with acute promyelocytic leukaemia. PMID- 9207424 TI - Increased plasma erythropoietin levels in lambs treated with parenteral iron. PMID- 9207425 TI - No sex-related difference in the myeloid:erythroid ratio in morphologically normal bone marrow aspirates. PMID- 9207426 TI - Prevalence of the FVQ506 (factor V Leiden) mutation in the normal and thrombophilic Algerian population. PMID- 9207427 TI - Sea-blue histiocytes in bone-marrow due to a long-term total parenteral nutrition including fat-emulsions. PMID- 9207428 TI - Liver biopsy in haemophilia. PMID- 9207429 TI - Liver biopsy in haemophilia. PMID- 9207430 TI - Genetic detection of factor V Leiden: the question of specificity. PMID- 9207431 TI - Homocysteine and thrombotic disease. PMID- 9207432 TI - Constitutive expression of Fas (Apo-1/CD95) ligand on multiple myeloma cells: a potential mechanism of tumor-induced suppression of immune surveillance. AB - The Fas (Apo-1/CD95) ligand (FasL) plays a central role in the elimination of target cells by effector T lymphocytes and in the suppression of cellular immune responses against nonmalignant and malignant cells. We show the expression of FasL on the surface of neoplastic plasma cells. We provide evidence that the FasL is functionally active because five of five neoplastic plasma cell lines tested killed CEM-C7H2 T-acute lymphoblastic leukemia (T-ALL) cells. The effect was mediated via the Fas (Apo-1/CD95) receptor molecule because blocking of Fas on the target cells or the FasL on the tumor cells by receptor- and ligand-specific monoclonal antibodies (MoAbs), respectively, protected T cells from being killed by myeloma cells. In addition, overexpression of the cowpox virus protein CrmA, a molecule with inhibitory potential on caspase-1 and caspase-8, specifically involved in Fas-induced signaling, protected T cells from being destroyed by the neoplastic cells or the agonistic anti-Fas MoAb. The potential of the malignant plasma cells to extinguish target T cells was independent of their own sensitivity to the agonistic anti-Fas MoAb, and FasL-positive (FasL+) CEM-C7H2 T cells were incapable of killing myeloma cells. Our results suggest that tumor cell-induced suppression of the immune system may be exerted via the FasL active on malignant plasma cells. Furthermore, loss of Fas expression or insensitivity to the agonistic anti-Fas MoAb do not seem to be prerequisites for myeloma cells to defeat T cells via Fas/FasL interaction. PMID- 9207433 TI - Bone marrow stroma-derived prolactin is involved in basal and platelet-activating factor-stimulated in vitro erythropoiesis. AB - Cooperation between in vitro exogenous prolactin (PRL), granulocyte-macrophage colony-stimulating factor (GM-CSF), and interleukin-3 (IL-3) at an early step of in vitro erythroid differentiation has been shown in a previous study. To gain more insight into the role of PRL in in vivo hematopoiesis, we have now addressed the involvement of endogenous PRL in the growth of hematopoietic progenitors in a bone marrow (BM) stroma environment. The possible modulation of local PRL production by the inflammatory mediator platelet-activating factor (PAF), which is known to be produced by BM cells and to regulate pituitary PRL release, has also been evaluated. Development of burst-forming unit-erythroid (BFU-E) colonies from CD34+ hematopoietic progenitors cultured on a BM stroma cells (BMSC) layer was slightly, but significantly, reduced in the presence of an anti-human PRL antibody. Pretreatment of BMSC with PAF increased the BFU-E colony efficiency of cocultured CD34+ cells, and this effect was completely abrogated by the antiserum. PAF-modulated release of PRL by BMSC was confirmed by an enzyme-linked immunospot (Elispot) technique. In addition, immunoprecipitation and Western blotting experiments showed two immunoreactive products in the BMSC culture medium. These corresponded to the nonglycosylated (23 kD) and glycosylated (25.5 kD) forms of pituitary PRL that are also expressed by the B-lymphoblastoid cell line IM9-P3. Specific increase of the nonglycosylated form and decrease of the glycosylated form was observed after PAF treatment. Polymerase chain reaction (PCR) amplification of reverse transcribed RNA using PRL-specific primers showed the presence of PRL message in BMSC and IM9-P3 cells. In situ hybridization experiments with a rat PRL cDNA probe cross-reacting with human PRL mRNA confirmed its presence in a small fraction of unstimulated BMSC and in the majority of PAF-stimulated BMSC. The enhancing effect of PAF on PRL-mediated colony formation, PRL release, and mRNA activation was counteracted by pretreating BMSC with the PAF-receptor (R) antagonist WEB 2170. Lastly, responsiveness of BMSC to PAF was substantiated by the presence of the PAF-R mRNA on these cells. PMID- 9207434 TI - Clinical features and treatment outcome of children with myeloid antigen positive acute lymphoblastic leukemia: a report from the Children's Cancer Group. AB - Leukemic cells from a significant number of children with acute lymphoblastic leukemia (ALL) express protein antigens characteristic of both lymphoid and myeloid cells, yet the clinical significance of this immunophenotype has remained controversial. In the current study, we have determined relationships between myeloid antigen expression and treatment outcome in a large cohort of children with newly diagnosed ALL. A total of 1,557 children enrolled on risk-adjusted Children's Cancer Group studies were classified as myeloid antigen positive (My+) or myeloid antigen negative (My-) B-lineage ALL (BL) or T-lineage ALL (TL), according to expression of CD7, CD19, CD13, and CD33 antigens on the surface of their leukemic cells. My+ patients in both BL and TL groups were more likely than My- patients to have favorable presenting features. Induction therapy outcome was similar for My+ and My- patients in both the BL and TL categories. Importantly, 4 year event-free survival (EFS) was similar for My+ BL (77.0%, standard deviation [SD] = 4.0%) versus My- BL (75.9%, SD = 1.8%) and for My+ TL (72.7%, SD = 7.1%) versus My- TL (70.1%, SD = 5.7%). An overall relative hazard rate (RHR) of 0.89 (P = .49) was determined by a cross strata analysis for My+ versus My- patients. Moreover, similar EFS and RHR also were found when My+ and My- BL patients were compared according to National Cancer Institute risk classification. Thus, patients with My+ ALL have similar treatment outcomes as My- ALL patients. In contrast to previous studies, this result was independent of treatment risk category, demonstrating that myeloid antigen expression was not an adverse prognostic factor for childhood ALL. PMID- 9207435 TI - Randomized trial of autologous filgrastim-primed bone marrow transplantation versus filgrastim-mobilized peripheral blood stem cell transplantation in lymphoma patients. AB - Although a large amount of data is available on the effects of filgrastim (granulocyte colony-stimulating factor [G-CSF]) on the mobilization of stem cells in the circulation, data concerning its effects on bone marrow (BM) harvesting is scarce and controversial. We have designed a randomized trial comparing filgrastim-mobilized peripheral blood stem cell (PBSC) transplantation with filgrastim-primed autologous bone marrow transplantation (ABMT). Fifty-five patients affected by non-Hodgkin's (n = 38) or Hodgkin's (n = 17) lymphoma, selected for autologous transplantation over a 12-month period in a single institution, were randomized 2:1 to undergo BM or PB harvest/collection after priming for 3 days with filgrastim, 16 microg/kg body weight daily subcutaneously. BM priming with G-CSF allowed the harvest of a significantly higher number of mononuclear cells (MNC) (0.53 x 10(8)/kg, range, 0.32 to 1.40), as compared with a historical control of unprimed BM harvests (0.43 x 10(8) MNC/kg, range, 0.15 to 0.72, P = .001). After high-dose ablative therapy, median time to neutrophil recovery above 0.5 x 10(9)/L was 12 days for BM and 11 days for PB (P = .219); median time to platelet recovery above 20 x 10(9)/L was 13 days for BM and 11 days for PB (P = .242). The same number of red blood cells, platelet transfusions, and posttransplant G-CSF doses were required in the two groups of patients. Less patients (50% v 70%) became febrile in the group transplanted with mobilized PB, but days of fever/patient and days on antibiotics were overlapping. The median time spent in the hospital after reinfusion was 16.5 and 15.5 days after primed BM and primed PB, respectively (P = .134). These data suggest that in patients with lymphoma submitted to autologous transplantation, the reinfusion of filgrastim-primed BM or filgrastim-mobilized PB leads to similar results, with an advantage of only 1 day in the neutrophil recovery and 1 day on the time spent in the hospital in favor of primed PB. Either option can be chosen on the basis of the availability of a surgery room or cell separator facilities and considering the patients' characteristics and wishes. PMID- 9207436 TI - Enzyme therapy in Gaucher disease type 1: effect of neutralizing antibodies to acid beta-glucosidase. AB - Gaucher disease type 1, a non-neuronopathic lysosomal storage disease, is caused by mutations at the acid beta-glucosidase locus. Periodic infusions of macrophage targeted acid beta-glucosidase reverse hepatosplenomegaly, hematologic, and bony findings in many patients. Two patients receiving enzyme therapy developed neutralizing antibodies to acid beta-glucosidase that were associated with a lack of improvement or progressive disease. After initial improvement, case 1 had no additional response to 2 years of high-dose (50 U/kg every 2 weeks) enzyme therapy. Similarly, case 2 initially showed a favorable response to enzyme therapy that plateaued after 1 year of treatment. Both patients developed minor allergic reactions and antibodies to acid beta-glucosidase within the first 6 months of treatment. Enzyme therapy was discontinued in case 1, with resultant disease progression and need for splenectomy. An immunosuppression/tolerization protocol was initiated in case 2 because of disease progression and stable neutralizing antibody titers. The IgG neutralizing antibodies rapidly and completely inactivated the wild-type, but not the N370S, acid beta-glucosidase in vitro. Antibodies to human serum albumin and chorionic gonadotropin also developed. The finding of neutralizing antibodies to acid beta-glucosidase during enzyme therapy for Gaucher disease has significant implications for monitoring the therapeutic responses and for potential alternative future therapies for Gaucher disease. PMID- 9207437 TI - Receptor protein tyrosine phosphatase gamma, Ptp gamma, regulates hematopoietic differentiation. AB - Murine embryonic stem (ES) cells have been a useful model system for the study of various aspects of hematopoietic differentiation. Because we had observed a sharp peak of expression of the receptor tyrosine phosphatase gamma (Ptp gamma) gene between 14 and 18 days of ES-derived embryoid body differentiation, we investigated the effect of perturbation of expression of the Ptp gamma gene on ES cell differentiation, first by analyzing the effect of Ptp gamma overexpression. The murine full-length Ptp gamma cDNA in an expression vector was transfected into ES-D3 cells and stably transfected clones were isolated. Ptp gamma was expressed as an approximately 230-kD cell surface protein, and differentiating ES clones that overexpressed Ptp gamma gave rise to a normal number of hematopoietic colonies, approximately 1 CFU per 100 cells. There was, however, a significant increase of expression of early hematopoietic markers in colonies from Ptp gamma overexpressing ES cells. To confirm that the pertubation of hematopoietic differentiation was a result of Ptp gamma overexpression, we isolated ES stem cell clones expressing Ptp gamma antisense constructs and assayed embryoid bodies for the presence of hematopoietic precursors. We observed a complete absence of methylcellulose colonies, indicating absence of hematopoietic lineages. Results of these experiments point to an essential role for Ptp gamma in hematopoietic differentiation. PMID- 9207438 TI - Prevention of thrombocytopenia by thrombopoietin in myelosuppressed rhesus monkeys accompanied by prominent erythropoietic stimulation and iron depletion. AB - The effectiveness of thrombopoietin (TPO) in alleviating thrombocytopenia was evaluated in a placebo-controlled study involving rhesus monkeys exposed to 5 Gy total-body irradiation (TBI) (300-kV x-rays) to result in 3 weeks of pancytopenia. Supraoptimal treatment with human recombinant TPO (10 microg/kg/d subcutaneously, days 1 to 21 after TBI) was highly effective in preventing thrombocytopenia, with nadirs for thrombocytes, on average, far higher than 100 x 10(9)/L, a greatly accelerated recovery to normal values, and no need for thrombocyte transfusions. TPO appeared to act selectively in that neutrophil regeneration was not influenced but red blood cell lineage recovery was prominently stimulated, with reticulocyte regeneration being initiated 10 days earlier than in placebo-treated animals. The reticulocytosis was followed by a normoblastosis that occurred earlier and was more pronounced than in placebo treated monkeys. The effect of TPO on the red blood cell lineage was also reflected in a less profound nadir for hemoglobin (Hb) and hematocrit values than in placebo controls. However, this effect was not followed by a rapid recovery to normal values, due to development of a microcytic hypochromic anemia. Iron depletion was demonstrated by measurements of total serum iron and total iron binding capacity (TIBC) and could be prevented by prophylactic intramuscular (IM) iron before TBI or corrected by IM iron after TPO treatment. Rechallenging with TPO in week 8 after TBI demonstrated a homogenous thrombocyte response similar in magnitude to the initial response, but a greatly diminished reticulocyte response. This demonstrated that the erythropoietic response to TPO administration depends on the hemopoietic state of the animal and may reflect multiple TPO target cells. It is postulated that the extremely rapid erythropoiesis due to TPO treatment in the initial regeneration phase following myelosuppression results in iron depletion by a mechanism similar to that seen following erythropoietin treatment in patients with end-stage renal failure. It is concluded that protracted TPO therapy to counteract thrombocytopenic states may result in iron depletion and that the iron status should be monitored before, during, and after TPO treatment. PMID- 9207439 TI - Selective expansion of primitive normal hematopoietic cells in cytokine supplemented cultures of purified cells from patients with chronic myeloid leukemia. AB - We have previously reported that primitive normal hematopoietic cells detectable as long-term culture-initiating cells (Ph-LTC-IC) are present at high levels in the blood of some patients with chronic myeloid leukemia (CML). We now show that this population can be expanded several-fold when highly purified CD34+CD38- cells isolated from the blood of such patients are cultured for 10 days in a serum-free medium containing 100 ng/mL of Flt3-ligand and Steel factor and 20 ng/mL of interleukin-3 (IL-3) and IL-6, and granulocyte colony-stimulating factor. In similar cultures initiated with CD34+CD38- cells from CML blood samples in which all of the LTC-IC were leukemic (Ph+), Ph+ LTC-IC activity was rapidly lost both in the presence and absence of admixed CD34+CD38- cells isolated from normal marrow. Conversely, the ability of normal LTC-IC to expand their numbers was shown to be independent of the presence of Ph+LTC-IC and later types of Ph+colony-forming cell (CFC) progenitors. In contrast to the LTC-IC, CFC were consistently amplified in cultures initiated with CML-derived CD34+CD38- cells and the additional CFC present after 10 days were, like the starting population of CFC, almost exclusively Ph+ regardless of the genotype(s) of the LTC-IC in the original CML samples. Amplification of the Ph+CFC population in these cultures showed the same factor dependence as previously demonstrated for the in vitro expansion of CFC from normal marrow CD34+CD38- cells. Ph+LTC-IC disappeared regardless of the cytokines present. Taken together these findings support a model of CML in which the leukemic stem cells are characterized by a decreased probability of self-renewal and an increased probability of differentiation. In addition, they suggest new opportunities for improving the treatment of CML using strategies that require autologous stem cell rescue. PMID- 9207440 TI - Expression of Wiskott-Aldrich syndrome protein (WASP) gene during hematopoietic differentiation. AB - The Wiskott-Aldrich syndrome (WAS) is an X-linked recessive disorder described as a clinical triad of thrombocytopenia, eczema, and immunodeficiency. The gene responsible for WAS encodes a 502-amino acid proline-rich protein (WASp) that is likely to play a role in the cytoskeleton reorganization and/or in signal transduction of hematopoietic cells. However, the function and the regulation of the WAS gene (WASP) have not yet been clearly defined. We have studied WASP expression at the transcriptional level in freshly isolated mature peripheral blood cells and during hematopoietic development. For this purpose, we have isolated CD34+ hematopoietic precursor cells from cord blood. These cells were cultured in vitro with various growth factors to generate committed or mature cells belonging to different hematopoietic differentiation pathways, such as granulocytic (CD15+) cells, monocytic (CD14+) cells, dendritic (CD1a+) cells, erythroid lineage (glycophorin A+) cells, and megakaryocytic cells (CD41+). We have shown by reverse transcriptase polymerase chain reaction analysis that the WASP transcript is ubiquitously detectable throughout differentiation from early hematopoietic progenitors, including CD34+CD45RA- and CD34+CD45RA+ cells, to cells belonging to different hematopoietic lineages, including erythroid committed and dendritic cells. In addition, Northern blot analysis showed that peripheral blood circulating lymphocytes (CD3+ and CD19+ cells) and monocytes express WASP mRNA. Several hematopoietic cell lines were tested and higher levels of expression were consistently detected in myelomonocytic cell types. By contrast, primary nonhematopoietic cells, including fibroblasts, endothelial cells, and keratinocytes, were consistently negative for WASP mRNA. PMID- 9207441 TI - Chronic expression of murine flt3 ligand in mice results in increased circulating white blood cell levels and abnormal cellular infiltrates associated with splenic fibrosis. AB - The effect of chronic expression of flt3 ligand (FL) on in vivo hematopoiesis was studied. Retroviral vector-mediated gene transfer was used in a mouse model of bone marrow transplantation to enforce expression of mouse FL cDNA in hematopoietic tissues. As early as 2 weeks posttransplantation, peripheral blood white blood cell counts in FL-overexpressing recipients were significantly elevated compared with controls. With the exception of eosinophils, all nucleated cell lineages studied were similarly affected in these animals. Experimental animals also exhibited severe anemia and progressive loss of marrow-derived erythropoiesis. All of the FL-overexpressing animals, but none of the controls, died between 10 and 13 weeks posttransplantation. Upon histological examination, severe splenomegaly was noted, with progressive fibrosis and infiltration by abnormal lymphoreticular cells. Abnormal cell infiltration also occurred in other organ systems, including bone marrow and liver. In situ immunocytochemistry on liver sections showed that the cellular infiltrate was CD3+/NLDC145+/CD11c+, but B220- and F4/80-, suggestive of a mixed infiltrate of dendritic cells and activated T lymphocytes. Infiltration of splenic blood vessel perivascular spaces resulted in vascular compression and eventual occlusion, leading to splenic necrosis consistent with infarction. These results show that FL can affect both myeloid and lymphoid cell lineages in vivo and further demonstrate the potential toxicity of in vivo treatment with FL. PMID- 9207442 TI - Engraftment and development of human CD34(+)-enriched cells from umbilical cord blood in NOD/LtSz-scid/scid mice. AB - Understanding the repopulating characteristics of human hematopoietic stem/progenitor cell fractions is crucial for predicting their performance after transplant into high-risk patients following high-dose therapy. We report that human umbilical cord blood cells, 78% to 100% of which express the hematopoietic progenitor cell surface marker CD34, can consistently engraft, develop, and proliferate in the hematopoietic tissues of sublethally irradiated NOD/LtSz scid/scid mice. Engraftment and development of CD34+ cells is not dependent on human growth factor support. CD34+ cells home to the mouse bone marrow (BM) that becomes the primary site of human hematopoietic development containing myeloid, lymphoid, erythroid, and CD34+ progenitor populations. Myeloid, and in particular lymphoid cells possessing more mature cell surface markers, comprise the human component of mouse spleen and peripheral blood, indicating that development proceeds from primary hematopoietic sites to the periphery. Repopulation of secondary recipients with human cells by BM from primary recipients demonstrates the maintenance of substantial proliferation capacity of the input precursor population. These data suggest that the cells capable of initiating human cell engraftment (SCID-repopulating cells) are contained in the CD34+ cell fraction, and that this mouse model will be useful for assaying the developmental potential of other rare human hematopoietic cell fractions in vivo. PMID- 9207443 TI - Selective expression of mRNA coding for the truncated form of erythropoietin receptor in hematopoietic cells and its decrease in patients with polycythemia vera. AB - The mRNA encoding full-length erythropoietin (EPO) receptor (EPOR-F) comprises exons I through VIII. Another membrane-bound EPOR (EPOR-T) isoform has a truncated cytoplasmic region and is encoded by the mRNA containing unspliced intron VII (EPOR-T mRNA). EPOR-T is believed to have a dominantly negative function against EPOR-F. We show that EPOR-T mRNA is markedly decreased in the blood cells of patients with polycythemia vera (PV). We also show that EPOR-T mRNA is not detected in erythroid/megakaryocytic leukemia cell lines, but is expressed in nonerythroid/nonmegakaryocytic lines, suggesting the presence of a cell type-specific system by which intron VII of the EPOR transcript is spliced. Deregulation of this splicing system in early hematopoietic progenitors possibly explains the profound decrease in EPOR-T mRNA and consequent pathophysiology of PV. PMID- 9207444 TI - Phenotypic consequences of mutations in the Fanconi anemia FAC gene: an International Fanconi Anemia Registry study. AB - Fanconi anemia (FA) is a genetically and phenotypically heterogeneous disorder defined by cellular hypersensitivity to DNA cross-linking agents; mutations in the gene defective in FA complementation group C, FAC, are responsible for the syndrome in a subset of patients. We have performed an analysis of the clinical effects of specific mutations in the FAC gene. Using the amplification refractory mutation system assays that we developed to rapidly detect FAC mutations, at least one mutated copy of the FAC gene was identified in 59 FA patients from the International Fanconi Anemia Registry (IFAR). This represents 15% of the 397 FA patients tested. FA-C patients were divided into three subgroups based on results of a genotype-phenotype analysis using the Cox proportional hazards model: (1) patients with the IVS4 mutation (n = 26); (2) patients with at least one exon 14 mutation (R548X or L554P) (n = 16); and (3) patients with at least one exon 1 mutation (322delG or Q13X) and no known exon 14 mutation (n = 17). Kaplan-Meier analysis shows that IVS4 or exon 14 mutations define poor risk subgroups, as they are associated with significantly earlier onset of hematologic abnormalities and poorer survival compared to exon 1 patients and to the non-FA-C IFAR population. There was no direct correlation between the degree of cellular hypersensitivity to the clastogenic effect of diepoxybutane and severity of clinical phenotype. Sixteen of the 59 FA-C patients (27%) have developed acute myelogenous leukemia. Thirteen of these patients have died; AML was the cause of death in 46% of the expired FA-C patients. This study enables us to define this clinically heterogeneous disorder genotypically to better predict clinical outcome and aid decision-making regarding major therapeutic modalities for a subset of FA patients. PMID- 9207445 TI - Functional and phenotypic characterization of cord blood and bone marrow subsets expressing FLT3 (CD135) receptor tyrosine kinase. AB - The class III receptor tyrosine kinase FLT3/FLK2 (FLT3; CD135) represents an important molecule involved in early steps of hematopoiesis. Here we compare cell surface expression of FLT3 on bone marrow (BM) and cord blood (CB) cells using monoclonal antibodies (MoAbs) specific for the extracellular domain of human FLT3. Flow cytometric analysis of MACS-purified BM and CB cells showed that 63% to 82% of BM CD34+ and 88% to 95% of the CB CD34+ cells coexpress FLT3. Clonogenic assays and morphological characterization of FACS-sorted BM CD34+ cells demonstrate that colony-forming unit-granulocyte-macrophage (CFU-GM) and immature myelo-monocytic precursor cells are enriched in the subpopulation staining most brightly with the FLT3 MoAb whereas the majority of the burst forming units-erythroid (BTU-E) and small cells with lymphoid morphology are found in the FLT3- population. In contrast, statistically indistinguishable proportions of CFU-granulocyte-erythrocyte-megakaryocyte-macrophage (CFU-GEMM) and more primitive cobblestone area forming cells (CAFC) were detected in both fractions, albeit the FLT3+ fraction consistently showed more CAFC activity than the FLT3- fraction. Although in both, BM and CB the majority of CD34+CD117+ (KIT+), CD34+CD90+ (Thy-1+), and CD34+CD109+ cells coexpress FLT3, three-color phenotypic analyses are consistent with the functional findings and suggest that the most primitive cells defined as CD34+CD38-, CD34+CD71low, CD34+HLA-DR-, CD34+CD117low, CD34+CD90+, and CD34+CD109+ express low levels of cell-surface FLT3 and were therefore not enriched to a statistically significant extent with the bright versus negative sorting scheme. Thus, clear segregation of the most primitive progenitors from BM CD34+ cells was confounded by low apparent levels of FLT3 cell-surface expression on these cells, whereas myeloid progenitors unambiguously segregated with the FLT3 brightest cells and erythroid progenitors with the FLT3 dimmest. Additional phenotypic analyses using MoAbs against progenitor/stem cell markers including the mucinlike molecule MGC-24v (CD164), the receptor tyrosine kinases TIE, FMS (CD115), and KIT (CD117) further illustrate the differences in surface antigen expression profiles of BM and CB CD34+ cells. Notably, CD115 is rarely detected on CB CD34+ cells, whereas 20% to 25% of the BM CD34+FLT3+ cells are CD115+. Furthermore, 80% to 95% of the CB CD34+CD117+ but only 60% to 75% of the BM CD34+CD117+ cells coexpress FLT3. Only a negligible amount of CD34+CD19+ are detected in CB, while in BM 20% to 30% of CD34+CD19+ presumed pro/pre-B cells coexpress FLT3. In contrast, the majority of the CD34+CD164+ and CD34+TIE+ subsets in both CB and BM coexpress FLT3. Analysis of unseparated cells showed that FLT3 expression is not restricted to CD34+ subsets. About 65% to 70% of lymphocyte-gated BM CD34-FLT3+ cells are positive for the monocytic marker CD115 whereas 25% to 30% of these cells consist of CD10 expressing B-cell precursors. Finally, CD34- monocytes in BM, CB, and PB express FLT3 whereas granulocytes are FLT3-. Our data show that detectable FLT3 appears first at low levels on the surface of primitive multilineage progenitor cells and disappears during defined stages of B-cell development, but is upregulated and maintained during monocytic maturation. PMID- 9207447 TI - Specific domains of fibronectin mediate adhesion and migration of early murine erythroid progenitors. AB - The binding of late stage erythroid cells to fibronectin (FN) has been well characterized and is believed to be critical for the terminal stages of erythroid differentiation, but the adhesive properties of more primitive murine erythroid progenitors and the role of these interactions during earlier stages of erythropoiesis has not been determined. Using chymotryptic fragments and inhibitory probes, we have tested the ability of each of the major cell binding domains of FN; the RGDS sequence, the CS-1 sequence, and the carboxy-terminal heparin-binding domain (HBD), to promote adhesion of primitive burst-forming unit erythroid (BFU-E), mature BFU-E, and colony-forming unit-erythroid (CFU-E). We found that only 10% to 15% of BFU-E bound to FN or to the RGDS sequence in contrast to 75% to 85% of CFU-E. Approximately 50% to 70% of BFU-E and 60% to 80% of CFU-E bound to the carboxy-terminal HBD and to the CS-1 sequence. The binding of BFU-E and CFU-E to the RGDS and CS-1 sites was blocked by beta1 integrin antibodies. These results suggest that binding to FN determinants is developmentally regulated during early erythroid differentiation. Erythroid progenitor migration within the bone marrow is thought to be important for the eventual release of reticulocytes into the circulation. A correlation between FN binding and the migratory capacity of erythroid cells has been suggested, although the ability of FN to promote migration of erythroid progenitors has not been directly measured. We measured migration of CFU-E on fragments of FN containing each cell binding region. CS-1-containing fragments, in addition to promoting adhesion of both BFU-E and CFU-E, supported the highest levels of CFU-E migration (11-fold above background). Migration was sixfold above background on intact FN and only threefold above background on RGDS-containing fragments. Fragments containing HBD alone, although they promoted adhesion of CFU-E, failed to support significant levels of migration. These results show that specific domains of FN possess distinct adhesion- and migration-promoting properties for murine erythroid progenitors. Regulation of the adhesive properties during erythroid differentiation may alter the ability of progenitors to migrate in the bone marrow and thus play an important role in normal murine erythroid differentiation. PMID- 9207446 TI - Features of macrophage differentiation induced by p19INK4d, a specific inhibitor of cyclin D-dependent kinases. AB - The mitogen-dependent induction of cyclin D-dependent kinase activity is required for cells to enter the DNA synthetic (S) phase of their division cycle. Immature 32Dcl3 myeloid cells (32D) proliferating in the presence of interleukin-3 (IL-3) normally express cyclins D2 and D3, which assemble into binary holoenzyme complexes with their catalytic subunits, CDK4 and CDK6. When 32D cells are switched to medium containing granulocyte colony-stimulating factor (G-CSF) instead of IL-3, D-type cyclins are degraded and, in the absence of their associated kinase activity, the cells arrest in the first gap phase (G1) of the cell cycle and differentiate to neutrophils. We derived 32D cells in which the expression of p19INK4d, a specific polypeptide inhibitor of CDK4 and CDK6, is regulated by the heavy metal-inducible sheep metallothionein promoter. Induction of p19INK4d in response to zinc prolonged cell survival in the absence of growth factor treatment. When maintained in medium containing both IL-3 and zinc, these cells lost cyclin D-dependent kinase activity, underwent G1 phase arrest, and acquired certain morphologic, antigenic, and functional properties of mononuclear phagocytes. Cells induced to express p19INK4d did not synthesize receptors for macrophage colony-stimulating factor (M-CSF/CSF-1) and reverted to an immature myeloid phenotype when shifted back into medium containing IL-3 alone. These cells exhibited accelerated differentiation to neutrophils in response to G-CSF but also gave rise to macrophage-like cells when maintained in medium containing both G-CSF and zinc. Therefore, the acquisition of macrophage properties in response to zinc treatment neither depended upon IL-3 nor upon G1 phase arrest per se and instead reflects some ability of p19INK4d, and presumably cyclin D dependent kinases, to affect myeloid differentiation. PMID- 9207448 TI - Downregulation of c-kit expression in human endothelial cells by inflammatory stimuli. AB - In recent studies we have shown that the expression of stem cell factor (SCF) in human endothelial cells is regulated by inflammatory processes. Gram-negative bacteria, interleukin-1 (IL-1), and lipopolysaccharide were able to upregulate the expression of SCF in human umbilical vein endothelial cells (HUVEC) (Blood 83:2836, 1994). Interestingly enough c-kit, the receptor of SCF, is coexpressed on HUVEC, suggesting an autoregulatory mechanism. To investigate the relation of c-kit and inflammatory processes we stimulated HUVEC with IL-1alpha and we established an in vitro model of inflammation. Binding experiments with 125I-SCF were performed to study the c-kit receptor expression on HUVEC. Scatchard analysis revealed both high-affinity receptors (K(d) approximately 0.36 nmol/L) and low-affinity receptors (K(d) approximately 2.9 nmol/L). Exposure to IL-1alpha led to a significant 50% reduction of c-kit high-affinity receptors, whereas the number of low-affinity receptors was not affected, in comparison to a control group of untreated HUVEC. Furthermore, using Northern blot analysis we studied the regulation c-kit mRNA expression in HUVEC after stimulation with IL-1alpha. Kinetic experiments showed a time-dependent downregulation of c-kit specific transcripts. In addition, we cocultured HUVEC with diverse bacterial strains. Experiments were performed over time with 1 x 10(6) bacteria/mL. Our data showed that, in contrary to the previously reported upregulation of SCF mRNA expression, stimulation with Yersinia enterocolitica or with Neisseria meningitidis led to a significant time-dependent downregulation of c-kit mRNA within 3 hours. These data indicate that inflammatory stimuli such as IL-1 or living bacteria activate a mechanism that downregulates c-kit receptor expression in human endothelial cells during the state of inflammation. PMID- 9207449 TI - Impaired generation of bone marrow B lymphocytes in mice deficient in C/EBPbeta. AB - CAAT/enhancer binding proteins (C/EBP) are a family of transcription factors that mediates adipocyte differentiation and the regulation of genes expressed in immune responses and inflammation, such as interleukin-6 (IL-6), IL-8, and granulocyte colony-stimulating factor (G-CSF). We investigated the role of C/EBPbeta (NF-IL6) in the generation of bone marrow B lymphocytes by taking advantage of C/EBPbeta-/- mice. We found that the expansion of bone marrow (BM) B lymphocytes was impaired in long-term lymphoid cultures from C/EBPbeta-/- mice. Consistent with this finding, the number of BM B cells was decreased in C/EBPbeta /- mice. Both the levels of IL-7 gene expression and bioactive IL-7 from BM stromal cells were decreased in C/EBPbeta-/- mice. Furthermore, the proliferative responsiveness of BM B-cell precursors to IL-7 was also reduced as compared to wild-type mice, indicating that C/EBPbeta is required for the generation of BM B cells induced by IL-7. Accordingly, IL-7 stimulates the C/EBPbeta DNA-binding activity of normal BM pre-B lymphocytes as well as of 70Z/3 pre-B cells. These results point to C/EBPbeta as a critical signaling molecule in BM B lymphopoiesis. PMID- 9207450 TI - Oncostatin M and leukemia inhibitory factor do not use the same functional receptor in mice. AB - Oncostatin M (OSM) and leukemia inhibitory factor (LIF) are members of the interleukin-6 (IL-6) subfamily of cytokines that use a common signal transducer gp130. Human OSM (hOSM) and LIF share a functional high-affinity receptor that is composed of gp130 and LIF receptor beta subunit (LIFRbeta). A second high affinity receptor for hOSM was recently found to be formed by gp130 and the hOSM receptor beta subunit. However, the nature of murine OSM (mOSM) and its receptors has remained unknown. Using the recently cloned mOSM cDNA, we produced recombinant mOSM and studied its biological activity and receptor structure. Murine hematopoietic cell lines M1 and DA1.a, an embryonic stem cell line CCE, and Ba/F3 transfectants expressing gp130 and LIFRbeta responded to murine LIF (mLIF) and hOSM equally well, while these cells responded to mOSM only at a 30 fold to 100-fold higher concentration than those of mLIF and hOSM. In contrast, NIH3T3 cells responded to mOSM, but not to mLIF and hOSM. Scatchard plot analyses showed that mOSM bound to gp130 with low-affinity (kd = 2.8 to 4.2 nmol/L) and that the binding affinity did not increase in the presence of LIFRbeta. However, mOSM bound to NIH3T3 cells with high-affinity (kd = 660 pmol/L), whereas mLIF did not bind to NIH3T3 cells at all. These results indicate that unlike hOSM, mOSM and mLIF do not share the same functional receptor, and mOSM delivers signals only through its specific receptor complex. Further studies in mice will define the physiological roles of OSM. PMID- 9207451 TI - Short- and long-term multilineage repopulating hematopoietic stem cells in late fetal and newborn mice: models for human umbilical cord blood. AB - Blood from late fetal and newborn mice is similar to umbilical cord blood obtained at birth in human beings, an important source of stem cells for clinical transplantation. The mouse model is useful because long-term functions can be readily assayed in vivo. To evaluate the functions of hematopoietic precursors in the blood and other tissues of late fetal and newborn mice, short- and long-term multilineage repopulating abilities were measured in vivo by competitive repopulation. Manipulations that might affect cell function, such as enrichment, tissue culture, or retroviral marking, were avoided. Hematopoietic stem cell functions of late fetal or newborn blood, liver, and spleen, were assayed as myeloid and lymphoid repopulating abilities relative to standard adult marrow cells. Donor cells from these tissues as well as adult control donor marrow cells were all of the same genotype. Cells from each donor tissue were mixed with portions from a pool of standard adult "competitor" marrow distinguished from the donors by genetic differences in hemoglobin and glucosephosphate isomerase. After 21 to 413 days, percentages of donor type myeloid and lymphoid cells in recipient blood were measured to assay the functional abilities of donor precursors relative to the standard. These relative measures are expressed as repopulating units, where each unit is equivalent to the repopulating ability found in 100,000 standard adult marrow cells. Thus, measures of repopulating units do not compare single cells but overall repopulating abilities of donor cell populations. Relative functional abilities in 1 million nucleated cells from late fetal or newborn blood were several times less than those found in adult marrow, but far more than in normal adult blood, and appeared to include long-term functional primitive hematopoietic stem cells (PHSC) similar to those in marrow. To estimate functional abilities of individual PHSC, variances among large groups of identical recipients were analyzed using both the binomial model and competitive dilution, a new model based on the Poisson distribution. The data best fit the hypothesis that individual PHSC from adult marrow, late fetal blood, or newborn blood each produce similar fractions of the total lymphoid and erythroid cells found in the recipient for many months. PMID- 9207452 TI - Telomerase regulation, cell cycle, and telomere stability in primitive hematopoietic cells. AB - Low levels of telomerase activity have recently been detected in human primitive hematopoietic cells, however, blood cells exhibit telomere shortening on cell proliferation. This challenging observation led us to study telomerase regulation and telomere length in human hematopoietic progenitor cells from fetal liver (FL), cord blood (CB), peripheral blood (PB), and bone marrow (BM). We found telomerase activity in CD34+/CD38+ cells exceeding levels in CD34+/CD38-, CD34- , and mononuclear cells (P < .05). Baseline telomerase activity was highest in BM (n = 5) CD34+ cells, followed by PB (n = 20), CB (n = 11), and FL (n = 1). Within 48 hours to 72 hours of in vitro culture of CD34+ cells in the presence of cytokines (KL, interleukin-3 [IL-3], IL-6, erythropoietin, granulocyte colony stimulating factor), telomerase activity was upregulated, peaked after 1 week of culture, and decreased to baseline levels or below detection after 3 to 4 weeks. Stimulation of CD34+ cells with single cytokines resulted in no or minor telomerase upregulation, whereas cytokine combinations resulted in a significant telomerase increase (P < .001). There was a correlation between telomerase activity, cell cycle status by BrdU incorporation, and induction of phosphorylated retinoblastoma protein, CDC2, CDK2, cyclin D1, and cyclin A, but not cyclin E and B1 after 72 hours with multiple (but not single) cytokines. In nonexpanding CD34+ cells, telomerase was low or undetectable. Secondary CD34+ cells showed a reduced ability to upregulate telomerase activity. Antiproliferative cytokines such as transforming growth factor-beta1 and high concentrations of all-trans-retinoic acid in cytokine-supported CD34+ cultures downmodulated telomerase activity. Average telomere lengths were 10.4 kbp, 7.4 kbp, and 7.6 kbp in CB, PB, and BM CD34+ cells, respectively. In ex vivo expansion cultures, an average telomeric DNA loss of 1 to 2 kbp over 4 weeks was observed. However, the rate of base pair loss per population doubling was significantly lower during the first 2 weeks, when telomerase was upregulated, than during weeks 3 and 4 of culture. In summary, telomerase is upregulated in response to cytokine-induced proliferation and cell cycle activation in primitive hematopoietic cells. Telomerase is downregulated between weeks 3 and 4 of ex vivo expansion culture linked with decreased proliferation and greater expansion of more mature cell subsets. Our data suggest that telomerase activity in hematopoietic cells reduces, but does not prevent, telomere shortening on proliferation. PMID- 9207453 TI - Characterization of cell cycle status and E2F complexes in mobilized CD34+ cells before and after cytokine stimulation. AB - Mobilized peripheral blood progenitors (CD34+ cells) have been shown to be either in the G0 or G1 phase of the cell cycle. In this study, it is shown that they are small cells with low protein content suggestive of G0. Support for this is provided by showing that the principal E2F complex consists of hypophosphorylated p130, E2F-4, and DP-1. The E2F-4 is more highly phosphorylated than in quiescent T cells. In response to cytokines in vitro, the CD34+ cells start to enter G1 within 8 hours and enter S-phase at about 48 hours. As cells enter G1, E2F-4 is dephosphorylated to several hypophosphorylated forms and three new DNA-binding complexes appear, including one containing E2F-4, DP-1, and p107. We suggest that mobilized CD34+ cells may be maintained in G0 by p130, E2F-4, and DP-1 and the coordinate dephosphorylation of E2F-4 and hyperphosphorylation of p130 may be central to the initiation of proliferation. PMID- 9207454 TI - Human plasminogen activator inhibitor-1 (PAI-1) deficiency: characterization of a large kindred with a null mutation in the PAI-1 gene. AB - Plasminogen activator inhibitor-1 (PAI-1), the primary inhibitor of tissue- and urokinase-type plasminogen activators, is considered a critical regulator of the fibrinolytic system. We previously reported a child with abnormal bleeding and complete PAI-1 deficiency caused by a frame-shift mutation in exon 4 of the PAI-1 gene. The purpose of this study was to provide genetic and clinical data on the extended pedigree of the original proband to better define the phenotype associated with PAI-1 deficiency. Allele-specific oligonucleotide hybridization was used to genotype individuals, and serum PAI-1 antigen was measured by enzyme linked immunosorbent assay. By this approach we have identified 19 individuals who are heterozygous for the PAI-1 null allele and 7 homozygous individuals with complete PAI-1 deficiency. Clinical manifestations of PAI-1 deficiency were restricted to abnormal bleeding, which was observed only after trauma or surgery in homozygous affected individuals. A spectrum of bleeding patterns was observed, including intracranial and joint bleeding after mild trauma, delayed surgical bleeding, severe menstrual bleeding, and frequent bruising. Fibrinolysis inhibitors, including epsilon-aminocaproic acid and tranexamic acid, were effective in treating and preventing bleeding episodes. Other than abnormal bleeding, no significant developmental or other abnormalities were observed in homozygous PAI-1-deficient individuals. Heterozygous PAI-1 deficiency was not associated with abnormal bleeding, even after trauma or surgery. These observations define the clinical spectrum of PAI-1 deficiency and provide additional evidence to support the hypothesis that the primary function of plasminogen activator inhibitor-1 in vivo is to regulate vascular fibrinolysis. PMID- 9207455 TI - Death of bystander cells by a novel pathway involving early mitochondrial damage in human immunodeficiency virus-related lymphadenopathy. AB - Destruction of immune cells in peripheral lymphoid tissues plays presumably a pivotal role in acquired immune deficiency syndrome pathogenesis. We found that cell suspensions obtained from lymph nodes of eight human immunodeficiency virus (HIV)-infected individuals contained variable proportions (2.1% to 18.3%, median 11.2%) of dead lymphocytes permeable to supravital dyes, represented by CD4+, CD8+, and B cells. The frequency of dead cells correlated directly (R = 0.847) with the amount of HIV provirus in the cell populations, and HIV provirus was enriched in the dead cell fractions. Similar proportions of dead cells were observed in cell suspensions from lymphadenopathic lymph nodes of HIV- donors, but not from small resting HIV- lymph nodes. Electron microscopic and flow cytometric analyses revealed that most dead cells from HIV+ lymph nodes lacked internucleosomal DNA fragmentation but displayed combined features of apoptosis and necrosis, eg, chromatin condensation and mitochondrial swelling. Cells with similar morphology were readily identified in lymph node tissue sections, and marked mitochondrial swelling could be occasionally observed in cells with otherwise normal morphology. Our findings have two major implications. One is that the in vivo cell death in HIV-infected lymph nodes occurs predominantly through a novel pathway, related to but distinct from classical apoptosis and characterised by early and severe mitochondrial damage. The second implication is that HIV-related lymphadenopathy is accompanied in vivo by massive destruction of uninfected lymph node cells. Comparable levels of cell death were observed in other inflammatory lymphadenopathies not related to HIV; however, the uniquely endless and generalized nature of HIV lymphadenopathy might render this "inflammatory" cell destruction a powerful pathogenetic mechanism, accounting for the progressive disruption and depletion of lymphoid tissues seen in HIV infection. PMID- 9207456 TI - HLA-DR-mediated signals for hematopoiesis and induction of apoptosis involve but are not limited to a nitric oxide pathway. AB - Cross-linking of major histocompatibility complex (MHC) class II antigens by anti HLA-DR monoclonal antibody (MoAb; H81.9; IgG2a) results in inhibition of hematopoiesis in canine and human models. Inhibition of hematopoiesis is associated with apoptosis in a proportion of marrow cells. Since in murine macrophages class II cross-linking triggers nitric oxide (NO) production, and NO is thought to affect regulation of hematopoiesis, we investigated whether NO was involved in our models. In murine J774 monocytes/macrophages, MoAb H81.9 did induce NO. NO production was blocked by N(G)-monomethyl-L-arginine (NMMA), an inhibitor of NO synthase (NOS), and by the antioxidant N-acetylcysteine (NAC). In human and canine long-term marrow cultures (LTMCs) and in enriched marrow monocytes, however, no measurable increase in NO production was noted after H81.9 exposure. Nevertheless, NAC protected LTMCs against H81.9 induced inhibition of hematopoiesis. Therefore, we determined the effect of an exogenous NO donator, sin-1 (3-morpholinosydnonimine), on canine and human LTMCs and on apoptosis. Sin 1 at concentrations > or =100 microg/mL inhibited LTMCs and induced apoptosis; at low concentrations (1 microg/mL), however, sin-1 stimulated the generation of colony-forming unit granulocyte-macrophage. Combined treatment with sin-1 at 100 microg/mL and MoAb H81.9 resulted in profound inhibition of hematopoiesis in both canine and human LTMCs, and had an additive effect on apoptosis. At 1 microg/mL sin-1 counteracted the effect of H81.9 on hematopoiesis. The effect of sin-1 on apoptosis and hematopoiesis in LTMC was largely prevented by NAC. These results are consistent with the hypothesis that HLA-DR mediated apoptosis and inhibition of hematopoiesis involve oxidative stress. However, the biphasic response of hematopoiesis to sin-1 suggests a complex regulatory network possibly related to differences in NO sensitivity of distinct subpopulations of cells. Signals in addition to NO appear to be involved in the effect of anti-HLA-DR MoAb on hematopoiesis. PMID- 9207457 TI - p100: a novel proliferation-associated nuclear protein specifically restricted to cell cycle phases S, G2, and M. AB - By immunization with nuclear lysates of L428 cells, we raised a monoclonal mouse antibody, Ki-S2 (IgG1). In Western blots, this antibody recognizes a nuclear antigen with an apparent molecular mass of 100 kD, termed p100. Protein sequencing of p100 showed that this is a hitherto unknown protein. Immunohistochemical examination of cryostat and paraffin sections of nearly all human tissue types and neoplasms showed that p100 was exclusively expressed in the nuclei of a fraction of proliferating cells. Cell sorting and fluorescence activated cell sorting analysis of stimulated peripheral blood mononuclear cells showed that p100 was exclusively expressed in proliferating cells from the transition G1/S until the end of cytokinesis. During mitosis, this protein is strictly associated with the spindle pole and with the mitotic spindle, whereas during S and G2, p100 is diffusely distributed throughout the cell nucleus. Immediately after completion of cytokinesis, p100 was rapidly degraded. In L428 cells, p100 is phosphorylated at least during mitosis. It has a turnover time of about 1 hour. Studies on routinely processed paraffin sections of specimens of malignant lymphoma, benign and malignant nevocellular tumors, and breast cancer showed that in all cases less than 40% of the Ki-67-positive growth fraction expressed p100. Thus, p100 might prove to be a more reliable measure of cellular proliferation and one that is more closely correlated to cancer prognosis, beyond its general biologic relevance as a cell cycle protein. PMID- 9207458 TI - Inhibition of apoptosis in a human pre-B-cell line by CD23 is mediated via a novel receptor. AB - Human CD23 is a 45-kD type II membrane glycoprotein, which functions as a low affinity receptor for IgE and as a ligand for the CD21 and CD11b/CD11c differentiation antigens. CD23 is released from the surface of cells as soluble fragments, and a 25-kD species of soluble CD23 (sCD23) appears to act as a multifunctional cytokine. In this report, sCD23 is shown to sustain the growth of low cell density cultures of a human pre-B-acute lymphocytic leukemia cell line, SMS-SB: no other cytokine tested was able to induce this effect. Flow cytometric analysis indicates that sCD23 acts to prevent apoptosis of SMS-SB cells. SMS-SB cells cultured at low cell density possess low levels of bcl-2 protein. Addition of sCD23 to cells at low cell density maintained bcl-2 expression at levels equivalent to those observed in SMS-SB cells cultured at higher cell densities. No CD23 mRNA was found in SMS-SB cells, ruling out an autocrine function for CD23 in this cell line model. Although SMS-SB cells do not express the known receptors for CD23, namely CD21, CD11b-CD18, or CD11c-CD18, the cells specifically bind CD23-containing liposomes, but not glycophorin-containing liposomes. Binding of CD23-containing liposomes is inhibited by anti-CD23 but not by anti-CD21 or anti CD11b/c monoclonal antibodies. The data show that sCD23 prevents apoptosis of the SMS-SB cell line by acting through a novel receptor. PMID- 9207459 TI - Prognostic significance of Bcl-2 protein expression and Bcl-2 gene rearrangement in diffuse aggressive non-Hodgkin's lymphoma. AB - The prognostic significance of Bcl-2 protein expression and bcl-2 gene rearrangement in diffuse large cell lymphomas (DLCL) is controversial. Bcl-2 protein expression prevents apoptosis and may have an important role in clinical drug resistance. The presence of a bcl-2 gene rearrangement in de novo DLCL suggests a possible follicle center cell origin and perhaps a distinct clinical behavior more akin to low-grade non-Hodgkin's lymphoma (NHL). The purpose of this study was to determine the impact of Bcl-2 protein expression and bcl-2 gene rearrangement (mbr and mcr) on survival of a cohort of patients with DLCL who were uniformly evaluated and treated with effective chemotherapy. Patients included the original MACOP-B cohort (n = 121) and the initial 18 patients treated with the VACOP-B regimen (total = 139). All patients had advanced-stage disease, were 16 to 70 years old, and corresponded to Working Formulation categories F, G, or H. No patients had prior treatment, discordant lymphoma, or human immunodeficiency virus seropositivity. Paraffin sections from diagnostic biopsies were analyzed for bcl-2 gene rearrangement including mbr and mcr breakpoints by polymerase chain reaction and Bcl-2 protein expression by immunohistochemistry. With a median follow-up of 81 months, overall (OS), disease free (DFS), and relapse-free survival (RFS) were measured to determine the prognostic significance of these parameters. Analyzable DNA was present in 118 of 139 (85%) cases, with 14 demonstrating a bcl-2 rearrangement (11 mbr, 3 mcr). All 14 of these bcl-2 gene rearrangement-positive cases were found in the 102 patients with a B-cell immunophenotype, but the presence of this rearrangement had no significant influence on survival. Bcl-2 protein expression was interpretable in 116 of 139 (83%) cases, with immunopositivity detected in 54 of 116 (47%). Using a cut-off of greater than 10% Bcl-2 immunopositive tumor cells for analysis, positive Bcl-2 protein expression was seen in 28 of 116 (24%) patients and the presence of this expression correlated with decreased 8-year OS (34% v 60%, P < .01), DFS (32% v 66%, P < .001), and RFS (25% v 59%, P < .001). Bcl-2 protein expression remained significant in multivariate analysis that included the clinical international prognostic index factors and immunophenotype (P < .02). In conclusion, although bcl-2 gene rearrangement status could not be shown to have an impact on outcome, Bcl-2 protein expression is a strong significant predictor of OS, DFS, and RFS in DLCLs. PMID- 9207460 TI - Recombinant toxins containing human granulocyte-macrophage colony-stimulating factor and either pseudomonas exotoxin or diphtheria toxin kill gastrointestinal cancer and leukemia cells. AB - The granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR) is a potential target for toxin-directed therapy, because it is overexpressed on many leukemias and solid tumors and apparently not on stem cells. To investigate the potential therapeutic use of GM-CSF toxins, we fused human GM-CSF to truncated forms of either Pseudomonas exotoxin (PE) or diphtheria toxin (DT) and tested the cytotoxicity of the resulting GM-CSF-PE38KDEL and DT388-GM-CSF on human gastrointestinal (GI) carcinomas and leukemias. Toward gastric and colon cancer cell lines, GM-CSF-PE38KDEL was much more cytotoxic than DT388-GM-CSF, with IC50s (concentration resulting in 50% inhibition of protein synthesis) of 0.5 to 10 ng/mL compared with 4 to 400 ng/mL, respectively. In contrast, toward leukemia lines and fresh bone marrow cells DT388-GM-CSF was more cytotoxic than GM-CSF PE38KDEL. The cytotoxicity of both GM-CSF-PE38KDEL and DT388-GM-CSF toward the human cells was specific, because it could be competed by an excess of GM-CSF. Binding studies indicated that human GM-CSF receptors were present on all of the human GI and leukemic cell lines tested, at levels of 540 to 3,700 sites per cell (kd = 0.2 to 2 nmol/L), and the number of sites per cell did not correlate with the cell type. A similar pattern of cytotoxicity was found with recombinant immunotoxins binding to the transferrin receptor, in that anti-TFR(Fv)-PE38KDEL was much more cytotoxic than DT388-anti-TFR(Fv) toward GI cells, but both were similar in their cytotoxic activity toward leukemia cells. The fact that PE is more effective than DT in killing GI but not leukemic tumor cells targeted by GM CSF indicates a fundamental difference in the way PE or DT gains access to the cytosol in these cells. GM-CSF-PE38KDEL and DT388-GM-CSF deserve further evaluation as possible treatments for selected tumors. PMID- 9207461 TI - Indomethacin is a potent inhibitor of pristane and plastic disc induced plasmacytomagenesis in a hypersusceptible BALB/c congenic strain. AB - Continuous indomethacin (INDO) administration in the drinking water (10 to 20 microg/mL) profoundly inhibited plasmacytoma (PCT) development initiated by three 0.2- or 0.5-mL intraperitoneal (i.p.) injections of pristane in hypersusceptible BALB/c.DBA/2-Idh1-Pep3 congenic mice. The most effective inhibitions were obtained with continuous INDO treatment. When treatment was delayed until 50 to 60 days after the first pristane injection, there was approximately a 50% reduction in PCT incidence. The primary action of pristane is the induction of a chronic inflammation in the peritoneal connective tissues and the formation of a microenvironment where PCTs develop. INDO, a powerful inhibitor of prostaglandin synthases (cyclooxygenases 1 and 2), did not inhibit the formation of mesenteric oil granuloma nor the appearance of cells in this chronic inflammatory tissue carrying c-myc illegitimately joined to an Ig heavy chain switch region, ie, the t(12;15) chromosomal translocation. INDO inhibited PCT induction by the i.p. implantation of 21 x 2 mm polycarbonate discs. These solid objects predominantly induce the formation of a patchy fibroplastic tissue on contacting peritoneal surfaces. These and previous data indicate that indomethacin inhibits an intermediate stage in PCT development after the arrival of cells bearing the T(12;15) translocation in the oil granuloma and before these cells acquire transplantability to a pristane-conditioned host. The biological mechanism that explains how INDO inhibits PCT development is not yet established but appears to result from decreased production of prostaglandins in chronic inflammatory tissues (oil granuloma, fibroplasia), suggesting that prostaglandins play an active role in oil and solid plastic induced PCT formation. PMID- 9207462 TI - Differential incorporation of ara-C, gemcitabine, and fludarabine into replicating and repairing DNA in proliferating human leukemia cells. AB - The major actions of nucleoside analogs such as arabinosylcytosine (ara-C) and fludarabine occurs after their incorporation into DNA, during either replication or repair synthesis. The metabolic salvage and DNA incorporation of the normal nucleoside, deoxycytidine, is functionally compartmentalized toward repair synthesis in a process regulated by ribonucleotide reductase. The aim of this study was to investigate the metabolic pathways by which nucleoside analogs that do (fludarabine, gemcitabine) or do not (ara-C) affect ribonucleotide reductase are incorporated into DNA in proliferating human leukemia cells. Using alkaline density-gradient centrifugation to separate repaired DNA from replicating DNA and unreplicated parental DNA strands, approximately 60% of ara-C nucleotide in DNA was incorporated by repair synthesis in CCRF-CEM cells; the remainder was incorporated by replication. In contrast, fludarabine and gemcitabine, nucleosides that inhibit ribonucleotide reductase and decreased deoxynucleotide pools, were incorporated mainly within replicating DNA. Hydroxyurea also depleted deoxynucleotide pools and increased the incorporation of ara-C into DNA by replicative synthesis. Stimulation of DNA repair activity by UV irradiation selectively enhanced the incorporation of all nucleosides tested through repair synthesis. These findings suggest that the pathways by which therapeutically useful nucleoside analogs are incorporated into DNA are affected by cellular dNTP pools from de novo synthesis and by the relative activities of DNA repair and replication. The antitumor activity of these drugs may be enhanced by combination with either ribonucleotide reductase inhibitors to increase their incorporation into replicating DNA or with agents that induce DNA damage and evoke the DNA repair process. PMID- 9207463 TI - Interleukin-6 overcomes p21WAF1 upregulation and G1 growth arrest induced by dexamethasone and interferon-gamma in multiple myeloma cells. AB - Interleukin-6 (IL-6) is a growth factor for multiple myeloma (MM) cells and can inhibit MM cell apoptosis. Our recent studies show that IL-6 facilitates MM cell growth via phosphorylation of retinoblastoma protein (pRB); however, the effects of IL-6 on those cyclins, cyclin-dependent kinases (CDKs), and CDK inhibitors (CDIs) that are known to regulate phosphorylation of pRB have not been defined in MM cells. In the present report, we cultured MM cell lines and patient cells with IL-6 and/or dexamethasone (Dex) and characterized changes in cell cycle; expression and association of cyclins, CDKs, and CDIs; and phosphorylation of pRB. Dex induced G1 growth arrest in MM cells, whereas IL-6 facilitated G1 to S phase transition; moreover, the effect of Dex was blocked by IL-6. p21WAF1 (p21) protein was constitutively expressed in the majority of MM cells independent of the status of p53. Its expression was upregulated by Dex and downregulated by IL 6; again, IL-6 inhibited the increase in p21 triggered by Dex. These alterations in p21 expression in MM cells were associated with changes in p21 binding to CDK2, CDK4, and CDK6; CDK2, CDK4, and CDK6 kinase activities; and phosphorylation of pRB. In contrast, expression of G1 cell cycle regulatory proteins, including p27KIP1, cyclin D2, and cyclin E, was not altered in MM cells cultured with Dex and/or IL-6. Finally, interferon-gamma (IFN-gamma) also induced G1 growth arrest and upregulated p21 protein expression; as with Dex, affects of IFN-gamma were inhibited by IL-6. Our results therefore show that changes in cell cycle distribution in MM cells triggered by Dex, IL-6, and IFN-gamma correlate with changes in p21 protein expression and implicate p21 in the coupling of Dex-, IL-6 , and IFN-gamma-related signals to G1 cell cycle regulation in MM cells. PMID- 9207464 TI - HLA-DR1-restricted bcr-abl (b3a2)-specific CD4+ T lymphocytes respond to dendritic cells pulsed with b3a2 peptide and antigen-presenting cells exposed to b3a2 containing cell lysates. AB - Chronic myeloid leukemia (CML) is characterized by a specific translocation of the c-abl oncogene on chromosome 9 to the break point cluster region (bcr) on chromosome 22, t(9;22) (q34;q11). This translocation results in the expression of a 210-kD bcr-abl protein fusion gene product. The juxtaposition of the bcr and abl genes produces a novel junctional amino acid sequence, which may be presented by antigen-presenting cells and recognized specifically by human T lymphocytes. We have generated a CD4+ T lymphocyte line (NG-1) which recognizes the peptide epitope (GFKQSSKALQR) in association with HLA-DRbeta1*0101-02. A comparison of antigen-presenting cells showed that CMRF-44+ blood dendritic cell presented a 12mer b3a2 peptide effectively. The b3a2 peptide was able to generate specific primary T-lymphocyte responses in other HLA-DR1 donors. We also show that bcr abl, b3a2 peptide-specific T-lymphocyte lines proliferate in response to bcr-abl b3a2 containing cell lysates (K562 or CML PBMC derived) but not control (including b2a2 CML PBMC) lysates. PMID- 9207465 TI - An experimental model of human leukemic meningitis in the nude rat. AB - An experimental animal model of meningeal leukemia was developed in the nude rat, rnu/rnu, using the human-derived acute lymphoblastic leukemia cell line HPB-ALL. Anesthetized rats were placed in a modified stereotaxic frame and then injected intrathecally, at the level of the cisterna magna, with human leukemic cells. Cerebrospinal fluid and tissue samples from brain, spinal cord, heart, liver, kidney, spleen, bone marrow, and cervical lymph nodes were subjected to histopathologic examination and molecular genetic screening by clonotype primer directed polymerase chain reaction (CPD-PCR). Ninety-three percent of animals (n = 14) developed signs of meningeal irritation leading to death 30 to 63 days postinjection (median, 36.0 days, mean, 38.7); death occurred between 30 and 39 days in 77% of all animals. Leukemic cells progressively infiltrated the pericerebellar and pericerebral subarachnoid space and infiltrated the Virchow Robin (perivascular) space. The infiltrating meningeal leukemia closely resembled the pathologic presentation in the human condition. By CPD-PCR, leukemic cells were first detected in cerebrospinal fluid (CSF) on day 4 postinjection, were variably present over the ensuing 17 days, and were consistently detected after day 21. At terminal stages, CPD-PCR tissue surveys showed leukemic DNA in all brains and spinal cords and rarely in cervical lymph nodes, but leukemic DNA was not detected in any other tissue screened. Leukemic meningitis was reliably produced with a predictable survival time. Intrathecal administration of leukemic cells was an efficient means of transmitting leukemic meningitis and it compartmentalized the disease to the central nervous system (CNS), eliminating potential complications of systemic illness. The use of human-derived cell lines may render this model more relevant to the development of future therapeutic strategies to treat leukemia and lymphoma that invade the CNS. PMID- 9207466 TI - RAR alpha1/RAR alpha2-PML mRNA expression in acute promyelocytic leukemia cells: a molecular and laboratory-clinical correlative study. AB - In addition to the major fusion gene PML-RAR alpha, the t(15; 17) in acute promyelocytic leukemia (APL) produces the reciprocal fusion gene RAR alpha-PML. To determine the scope of RAR alpha-containing mRNA expression in APL cells, we tested PML-RAR alpha-positive APL cells for the presence of mRNAs initiated from two distinct RAR alpha gene promoters, alpha1 and alpha2. From the normal allele, both RAR alpha1 and RAR alpha2 mRNAs were expressed in all APL cases (N = 24). From the translocated allele, RAR alpha1-PML mRNA was expressed in 77% and RAR alpha2-PML mRNA in 28% of cases (N = 98). RAR alpha2-PML mRNA was not observed in the absence of RAR alpha1-PML mRNA. There was no association between RAR alpha1 PML or RAR alpha2-PML mRNA expression and the type of PML-RAR alpha mRNA formed by either 5' or 3' breaksites in the PML gene. RAR alpha1-PML mRNAs and RAR alpha2-PML mRNAs from 5' PML breaksite cases coded for full-length RAR alpha-PML proteins but RAR alpha2-PML mRNAs from 3' PML breaksite cases encoded a truncated RAR alpha2 peptide. RAR alpha1/alpha2-PML mRNA expression was not associated with differences in APL cell sensitivity to all-trans retinoic acid(tRA)-induced differentiation in vitro or in clinical outcome after tRA or chemotherapy induction therapy (protocol E2491). Our analysis indicated that RAR alpha1/alpha2 PML mRNA expression markedly differs from normal RAR alpha1/alpha2 mRNA expression, that the difference in RAR alpha1-PML and RAR alpha2-PML mRNA expression frequency is primarily related to the genomic separation of the RAR alpha1 and RAR alpha2 coding exons, and that variations in RAR alpha1/alpha2-PML mRNA expression likely have no clinically relevant function in APL cells. PMID- 9207467 TI - Molecular profile of Epstein-Barr virus in human immunodeficiency virus type 1 related lymphadenopathies and lymphomas. AB - Human immunodeficiency virus type 1 (HIV-1)-infected patients develop a spectrum of lymphoproliferative disorders ranging from nonneoplastic lymphadenopathies to B-cell lymphomas. Although evidence suggests that Epstein-Barr virus (EBV) might be involved, its molecular profile and expression pattern in HIV-1-related lymphoproliferations remain to be defined. Using polymerase chain reaction-based techniques, we studied EBV types and variants in 28 lymphadenopathy lesions and in 20 lymphomas (15 large cell and 5 Burkitt-like). EBV was detected in 89% of lymphadenopathies and in 80% of lymphomas; viral DNA content was significantly higher in the latter. EBNA2 and LMP1 gene analysis indicated that half of the EBV+ lymphadenopathies were coinfected with both EBV type 1 and 2 strains and/or multiple type 1 variants. Conversely, all but one lymphoma carried a single viral variant, consistently type 1 in large cell lymphomas, and type 2 in Burkitt-like tumors. Most lymphomas, but no lymphadenopathies, showed monoclonal Ig heavy chain rearrangement. Analysis of 5 large cell lymphomas and 9 lymphadenopathies for EBV transcripts identified LMP1 mRNA in most samples, and the EBNA2 transcript in all tumors. These findings provide evidence of a heterogeneous EBV population in lymphadenopathy lesions, strengthen the notion that lymphomas arise from clonal expansion of EBV+ cells, and suggest different roles for EBV types 1 and 2 in HIV-1-related lymphoproliferations. PMID- 9207468 TI - Sequence analysis of the Epstein-Barr virus (EBV) latent membrane protein-1 gene and promoter region: identification of four variants among wild-type EBV isolates. AB - Sequence variations in the Epstein-Barr virus (EBV) encoded latent membrane protein-1 (LMP-1) gene have been described in a Chinese nasopharyngeal carcinoma derived isolate (CAO), and in viral isolates from various EBV-associated tumors. It has been suggested that these genetic changes, which include loss of a Xho I restriction site (position 169425) and a C-terminal 30-base pair (bp) deletion (position 168287-168256), define EBV genotypes associated with increased tumorigenicity or with disease among particular geographic populations. To determine the frequency of LMP-1 variations in European wild-type virus isolates, we sequenced the LMP-1 promoter and gene in EBV from lymphoblastoid cell lines from healthy carriers and patients without EBV-associated disease. Sequence changes were often present, and defined at least four main groups of viral isolates, which we designate Groups A through D. The widespread prevalence of LMP 1 sequence variations, particularly the Xho I polymorphism and the 30-bp deletion, indicate that they cannot be used as simple markers for oncogenic viruses related to particular forms of EBV-associated tumor. Several of the structural changes detected occur, however, at sites where they may affect transcription, translation, or function of LMP-1. Future in vitro studies should aim to establish the functional importance of variations at these sites. PMID- 9207469 TI - Antisense oligodeoxyribonucleotides suppress hematologic cell growth through stepwise release of deoxyribonucleotides. AB - Antisense oligodeoxyribonucleotides (ODNs) are now being extensively investigated in an attempt to achieve cell growth suppression through specific targeting of genes related to cell proliferation, despite increasing evidence of non-antisense cytotoxic effects. In the context of anti-BCR/ABL antisense strategies in chronic myeloid leukemia, we have reexamined the antiproliferative effect of phosphodiester and phosphorothioate ODNs on the leukemic cell line BV173 and on CD34+ bone marrow cells in liquid culture. The 3' sequences of the ODNs determine their effect. At concentrations of 10 micromol/L (for phosphorothioate ODNs) or 25 micromol/L (for phosphodiester ODNs), all the tested ODNs exert an antiproliferative activity, except those that contain a cytosine residue at either their two most terminal 3' positions. We show that this antiproliferative effect is due to the toxicity of the d-NMPs (5' monophosphate deoxyribonucleosides), the enzymatic hydrolysis products of the ODNs in culture medium. The toxicity of the d-NMPs on hematologic cells depends on their nature (d-CMP [2'deoxycytidine 5'-monophosphate] is not cytotoxic), on their concentration (d-GMP [2'-deoxyguanosine 5'-monophosphate], TMP [thymidine 5' monophosphate], and d-AMP [2'-deoxyadenosine 5'-monophosphate] are cytotoxic at concentrations between 5 and 10 micromol/L), and on the coincident presence of other d-NMPs in the culture medium (d-CMP neutralizes the toxicity of d-AMP, d GMP, or TMP). The antiproliferative activity of ODNs is thus restricted to conditions where the 3' hydrolysis process by exonucleases generates significant amounts of d-NMPs with a low proportion of d-CMP. Our results reveal a novel example of a nonantisense effect of ODNs, which should be taken into account when performing any experiment using assumed antisense ODNs. PMID- 9207470 TI - Circulating blood B cells in multiple myeloma: analysis and relationship to circulating clonal cells and clinical parameters in a cohort of patients entered on the Eastern Cooperative Oncology Group phase III E9486 clinical trial. AB - Recent analyses of circulating blood B cells in myeloma have generated controversy concerning the exact levels of these cells and whether they may represent circulating clonal tumor B cells. Previous reports suggested that CD19+ B cells are markedly increased in myeloma patients and that this population shares clonotypic rearrangements with the malignant plasma cell. We studied the numbers of CD19+ B cells by flow cytometry in previously untreated newly diagnosed myeloma patients in Eastern Cooperative Oncology Group (ECOG) phase III trial E9486. There were 628 patients who were eligible for the clinical protocol E9486, but of these 521 were also entered on the companion laboratory study (E9487) and had CD19 data. In comparison with normal controls, the myeloma patients exhibited a marked heterogeneity in the number of circulating CD19+ B cells as detected by flow cytometry. Approximately 20% of patients had significantly increased levels of circulating CD19+ B cells. However, the total CD19+ blood population from myeloma was not significantly different from the median of age-matched, normal controls. Analysis of CD19+ blood cells in relationship to circulating clonal cells was done in 13 myeloma patients using a clonotypic, quantitative allele-specific oligonucleotide-polymerase chain reaction (PCR) assay. No correlation was found between the numbers of CD19+ B cells (range, 5% to 51%) and PCR estimates of the number of clonal cells in the peripheral blood (range, .009% to 3.6%). Low CD19+ B-cell level (<125 microL) was associated with clinical stage III (P = .033). A significant relationship exists between higher levels (> or = 125/microL) of CD19 cells and longer overall survival (P < .0001). In addition, high CD19 levels also predicted a clinical response and longer event-free survival. There was a strong inverse association between the level of CD19 values at diagnosis and infections within the first 2 months of diagnosis. Importantly, the number of deaths related to infections was significantly greater in the low versus high CD19 group (P < .0202). Also, CD19 is an independent prognostic factor in addition to plasma cell labeling indices, beta2-microglobulin, hemoglobin, and plasmablastic morphology. Patients with infections were more likely to have low levels of CD19+ cells. In summary, higher CD19+ cell levels are a favorable prognostic sign with no apparent relationship to circulating tumor cells. In addition, this analysis strongly suggests that low peripheral blood levels of CD19+ cells are an adverse prognostic sign in myeloma. The CD19+ cell levels in myeloma patients is an important parameter in the overall assessment of these patients. PMID- 9207471 TI - Sensitivity of acute leukemia cells to cytarabine is a correlate of cellular es nucleoside transporter site content measured by flow cytometry with SAENTA fluorescein. AB - Cytarabine (araC) is converted to araC 5'-triphosphate after entering leukemia cells as a substrate for nucleoside transport processes. This study tested the relationship between araC cytotoxicity, measured in an in vitro tetrazolium dye reduction assay of cell viability, and the cellular abundance of es nucleoside transport elements, assayed by a flow cytometric method that used the es-specific stain, 5-(SAENTA-x8)-fluorescein (5-(Sx8)-F), in cultured leukemia cells and in myeloblasts and lymphoblasts (blasts) from leukemia patients. Cellular es site abundance (B(max) value for 5-(Sx8)-F binding) varied sixfold among nine leukemic myeloblast samples from patients. In cultured OCI/AML-2 myeloblasts and CCRF-CEM T-lymphoblasts, and in fresh leukemic blasts, es sites were fractionally blocked by treatment with graded concentrations of nitrobenzylthioinosine (NBMPR), an inhibitory es site ligand, to simulate the variation in es expression found in leukemic blasts from patients with acute myeloid leukemia. When the cytotoxicity of a single concentration of araC was determined in NBMPR-treated leukemia cells, cell kill correlated closely with the intensity of 5-(Sx8)-F fluorescence (r = .92 to .99), a measure of the cell surface abundance of functional es nucleoside transporter sites. Concentrations of NBMPR that achieved half-maximal reduction (4.3 to 12 nmol/L) of cellular 5-(Sx8)-F fluorescence (measured by flow cytometry) approximated IC50 values (1 to 10 nmol/L) previously found for inhibition by NBMPR of es-mediated nucleoside fluxes in several cell types, supporting the view that 5-(Sx8)-F interacted with the estransporter. The correlation of araC cytotoxicity and the B(max) for 5-(Sx8)-F binding to es sites in cultured leukemia cells and in leukemic blasts from acute leukemia patients (r = .95) suggests that the flow cytometry assay of es capacity may be useful in predicting clinical response to araC. PMID- 9207472 TI - EORTC classification for primary cutaneous lymphomas: a proposal from the Cutaneous Lymphoma Study Group of the European Organization for Research and Treatment of Cancer. AB - Primary cutaneous lymphomas represent a heterogeneous group of T- and B-cell lymphomas that show considerable variation in histology, phenotype, and prognosis. Recently, the European Organization for Research and Treatment of Cancer (EORTC) Cutaneous Lymphoma Project Group has reached consensus on a new classification for this group of diseases. The EORTC classification for primary cutaneous lymphomas is based on a combination of clinical, histologic, and immunophenotypic criteria, and thus contains well-defined disease entities rather than histologic subgroups. In addition, this new classification contains a number of provisional entities, which may display characteristic histologic features, but are not yet well defined clinically. These provisional entities account for less than 5% of all primary cutaneous lymphomas. In this report the basic principles of this new classification, as well as the characteristic features of the different disease entities, are described. In addition, survival data of 626 patients with primary cutaneous lymphomas derived from the registry of the Dutch Cutaneous Lymphoma Working Group, illustrating the clinical validity of this new classification, are presented. PMID- 9207473 TI - Calreticulin biosynthesis and processing in human myeloid cells: demonstration of signal peptide cleavage and N-glycosylation. AB - Calreticulin is a soluble endoplasmic reticulum protein comprising the major storage reservoir for inositol trisphosphate-releasable calcium. Although its highly conserved primary structure and a wide range of functions have been well described, less attention has been paid to its biosynthesis, particularly in human tissues. We report analyses of synthesis, proteolytic processing and glycosylation of human calreticulin. In both HL-60 and PLB-985 myeloid cell lines calreticulin was immunoprecipitated as a single 60-kD species without evidence of precursor forms. However, in vitro cell-free synthesis produced a 62-kD primary translation product, which in the presence of microsomal membranes, was processed by cotranslational signal peptide cleavage to a 60-kD species that comigrated with mature calreticulin produced in myeloid cells. Neither tunicamycin treatment of the cells nor endoglycosidase digestion of calreticulin resulted in any forms other than the 60-kD protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis, suggesting that the potential site for N-glycosylation at asparagine-327 was unmodified. However, oxidative derivatization of carbohydrate components with digoxigenin showed that human calreticulin produced in either HL-60 cells or Sf9 insect cells is glycosylated, indicating that glycosylated and nonglycosylated human calreticulin have indistinguishable electrophoretic mobilities. Direct measurement by phenol-H2SO4 confirmed the presence of carbohydrate on recombinant human calreticulin. These data show that human myeloid calreticulin undergoes cotranslational signal peptide cleavage and posttranslational N-linked glycosylation. Although glycosylation of calreticulin has been shown in rat liver and bovine liver and brain, it has been reported to be lacking in other tissues including human lymphocytes. PMID- 9207474 TI - Mouse mast cells that possess segmented/multi-lobular nuclei. AB - Because in humans mast cells and basophils tend to possess nonsegmented and segmented/multi-lobular nuclei, respectively, nuclear morphology has been a major criterion for assessing the lineage of metachromatic cells of hematopoietic origin. Immature metachromatic cells with mono- and multi-lobular nuclei were both obtained when bone marrow cells from BALB/c mice were cultured for 3 weeks in the presence of interleukin-3. Analogous to the indigenous mature mast cells that reside in the peritoneal cavity and skin, both populations of in vitro derived cells expressed the surface receptor c-kit, the chymase mouse mast cell protease (mMCP) 5, the tryptase mMCP-6, and the exopeptidase carboxypeptidase A (mMC-CPA). Immunogold electron microscopy confirmed the granule location of mMC CPA and mMCP-6 in both populations of cells, and cytochemical analysis confirmed the presence of chymotryptic enzymes in the granules. Because mature mast cells possessing multi-lobular nuclei also were occasionally found in the skeletal muscle and jejunum of the BALB/c mouse, the V3 mouse mast cell line was used to investigate the developmental relationship of mast cells that have very different nuclear structures. After the adoptive transfer of V3 mast cells into BALB/c mice, v-abl-immortalized mast cells with mono- and multi-lobular nuclei were detected in the lymph nodes and other tissues of the mastocytosis mice that expressed c-kit, mMCP-5, mMCP-6, and mMC-CPA. These studies indicate that mouse mast cells can exhibit varied nuclear profiles. Moreover, the nuclear morphology of this cell type gives no insight as to its protease phenotype or stage of development. PMID- 9207475 TI - Alternative splicing of a novel glycophorin allele GPHe(GL) generates two protein isoforms in the human erythrocyte membrane. AB - The Henshaw antigen (synonym: He or MNS6) is carried by an altered form of glycophorin B (GPB), but the molecular basis for its variable expression or quantitative polymorphism remains largely undefined. We report here the identification and analysis of a novel glycophorin He allele, GPHe(GL), which gives rise to the expression of two protein isoforms in the erythrocyte membrane. In addition to the nucleotide changes defining the epitopic sequence of He, a single C-to-G nucleotide transversion in exon V coding for the membrane domain was found to cause aberrant RNA splicings by creating a new acceptor splice site. In addition, a T-to-G transversion at -6 position of the acceptor splice site for exon IV was identified. Both full-length and truncated transcripts of GPHe(GL) were detected as the result of partial activation of the new acceptor splice site and partial inactivation of the normal splice sites. The full-length cDNA encoded He, S, and U antigens, whereas the three truncated ones lacked either the sequence for S and U antigens or a large portion of the membrane domain or both. The GPB gene on the other chromosome was apparently normal and its transcript encoded N, s, and U antigens. These results correlate alternative RNA splicing with the expression of two GPHe isoforms and thus delineate a new mechanism for the phenotypic diversity of membrane glycophorins. PMID- 9207477 TI - Sensitivity to a metabolite of diclofenac as a cause of acute immune hemolytic anemia. AB - A 75-year-old woman taking the nonsteroidal anti-inflammatory drug diclofenac (DCF) presented with acute Coombs-positive hemolytic anemia and subsequently developed renal failure. A drug-dependent antibody specific for red blood cells (RBCs) could not be demonstrated by in vitro testing with DCF. However, her serum was found to contain an IgM antibody that reacted strongly with RBCs in the presence of urine from any of four subjects who had ingested DCF. The active substance in urine was isolated, subjected to high-performance liquid chromatographic (HPLC) analysis, and found to be a glucuronide conjugate of a known DCF metabolite, 4'-hydroxydiclofenac (4'-OH DCF). Negative results were obtained with four other DCF metabolites. Two 4'-OH DCF glucuronides were synthesized in vitro using a liver microsomal system. One promoted agglutination of normal RBCs by the patient's serum and was identified as the glucuronide ester of 4'-OH DCF by proton nuclear magnetic resonance (NMR) analysis. Studies with a panel of RBCs showed that the patient's antibody reacted preferentially with the e antigen of the Rh system. Acute immune hemolytic anemia in this patient appears to have been caused by sensitization to DCF modified by 4' hydroxylation and glucuronidation. This is the first reported example of immune cytopenia caused by sensitivity to a glucuronide conjugate of a drug metabolite. Since glucuronidation is a common pathway of drug metabolism, studies of the frequency with which glucuronide derivatives of primary medications cause immune cytopenia seem warranted. PMID- 9207476 TI - Characterization of the underlying molecular defect in hereditary spherocytosis associated with spectrin deficiency. AB - Several subsets of patients with hereditary spherocytosis (HS) have been defined based on the specific red blood cell membrane protein deficiencies involving spectrin, ankyrin, band 3, and protein 4.2. Mutations of the genes encoding these proteins are currently being uncovered. Regarding spectrin, only three isolated cases of beta-spectrin gene mutations were recently reported in association with HS and spectrin deficiency. We have screened the coding region of the beta spectrin gene using the SSCP technique, in 40 families with HS associated with spectrin deficiency or combined spectrin and ankyrin deficiencies. In this report we describe six frameshift and nonsense mutations and four missense mutations of the beta-spectrin gene in 11 unrelated families. Taking advantage of modifications in the restriction enzyme recognition sequences introduced by the mutations, we show, in all cases of frameshift and nonsense mutations, the loss of heterozygosity at the cDNA level when compared to genomic DNA, reflecting the absence of the mutant mRNA transcripts. In one family with a large pedigree including six generations and 112 members, we firmly establish the autosomal dominant inheritance of one of the beta-spectrin null mutations. Most of the mutations described are responsible for a phenotype of mild to moderate autosomal dominant form of HS associated with a conspicuous spherocytosis with frequent spiculated cells (8% to 15% acanthocytes). One missense mutation appears to be associated with a recessive form of the disease. Five common restriction enzyme polymorphisms of the coding region of the beta-spectrin gene are also described. Overall, these findings underscore the importance of the beta-spectrin gene mutations in the pathogenesis of HS and reemphasizes the extreme heterogeneity of the underlying molecular basis of this condition. PMID- 9207478 TI - Modulation of clinical expression and band 3 deficiency in hereditary spherocytosis. AB - We present two novel alleles of the anion-exchanger 1 (AE1) gene, allele Coimbra and allele Mondego. Allele Coimbra (V488M, GTG --> ATG) affects a conserved position in the putative second ectoplasmic loop of erythrocyte band 3. In 15 simple heterozygotes, it yielded a mild form of hereditary spherocytosis (HS) with band 3 deficiency (-20% +/- 2%) and a reduced number of 4,4'-diisothiocyano 1,2-diphenylethane-2,2'-disulfonate (H2DIDS) binding sites (-35%). However, two additional heterozygotes presented with an aggravated HS and a more pronounced reduction of band 3 (-40%) and of H2DIDS binding sites (-48%). They carried, in trans to allele Coimbra, allele Mondego, defined by two mutations: E40K, GAG --> AAG, the known mutation Montefiore, and P147S, CCT --> TCT, a novel mutation, both located in the cytoplasmic domain of band 3. Allele Mondego itself resulted in no clinical or hematologic HS signs in the simple heterozygous state. Yet it yielded a slight decrease in band 3 (-6% to -12%) and in the number of H2DIDS binding sites (-19%). Thus, the more pronounced decrease in band 3 in the two compound heterozygotes derived from the additive effects of two unequally expressed AE1 alleles, resulting in a more severe clinical picture. PMID- 9207479 TI - Restoration of the CCAAT box or insertion of the CACCC motif activates [corrected] delta-globin gene expression. AB - Hemoglobin A2 (HbA2), which contains delta-globin as its non-alpha-globin, represents a minor fraction of the Hb found in normal adults. It has been shown recently that HbA2 is as potent as HbF in inhibiting intracellular deoxy-HbS polymerization, and its expression is therefore relevant to sickle cell disease treatment strategies. To elucidate the mechanisms responsible for the low-level expression of the delta-globin gene in adult erythroid cells, we first compared promoter sequences and found that the delta-globin gene differs from the beta globin gene in the absence of an erythroid Kruppel-like factor (EKLF) binding site, the alteration of the CCAAT box to CCAAC, and the presence of a GATA-1 binding site. Second, serial deletions of the human delta-globin promoter sequence fused to a luciferase (LUC) reporter gene were transfected into K562 cells. We identified both positive and negative regulatory regions in the 5' flanking sequence. Furthermore, a plasmid containing a single base pair (bp) mutation in the CCAAC box of the delta promoter, restoring the CCAAT box, caused a 5.6-fold and 2.4-fold (P < .05) increase of LUC activity in transfected K562 cells and MEL cells, respectively, in comparison to the wild-type delta promoter. A set of substitutions that create an EKLF binding site centered at -85 bp increased the expression by 26.8-fold and 6.5-fold (P < .05) in K562 and MEL cells, respectively. These results clearly demonstrate that the restoration of either an EKLF binding site or the CCAAT box can increase delta-globin gene expression, with potential future clinical benefit. PMID- 9207480 TI - Genetic heterogeneity in heterocellular hereditary persistence of fetal hemoglobin. AB - A large English pedigree in which heterocellular hereditary persistence of fetal hemoglobin (HPFH) segregates is described. beta-globin cluster deletions and gamma gene promoter mutations associated with HPFH have been excluded. Of particular importance in this pedigree is the absence of any cosegregating hemoglobinopathy, thus allowing observation of the segregation pattern of this form of HPFH without the complicating effect of a beta-globin gene mutation. Information gained in this study confirms that the extent of elevation of hemoglobin (Hb) F and F cells varies between affected individuals. There are one example of incomplete penetrance and three examples of father-to-son transmission, thus excluding X-linked inheritance. Consistent with previous reports, the most likely mode of inheritance is autosomal codominant. Linkage studies using a beta-globin cluster microsatellite show no evidence of linkage to this chromosomal region implicating the presence of trans-acting regulatory factor(s). We have recently mapped one such locus to the chromosome 6q region in a very large Asian-Indian pedigree. Linkage to chromosome 6q in the English pedigree was excluded, thus indicating the presence of genetic heterogeneity in heterocellular HPFH. PMID- 9207481 TI - Relationship between the phenotypes of circulating erythrocytes and cultured erythroblasts in paroxysmal nocturnal hemoglobinuria. AB - To investigate erythropoiesis in paroxysmal nocturnal hemoglobinuria (PNH), we studied the expression of glycosylphosphatidylinositol (GPI)-anchored membrane proteins on circulating erythrocytes and erythroblasts obtained by erythropoietic cell culture in nine patients with this disease. One-color and two-color flow cytometric analyses were performed using monoclonal antibodies for decay accelerating factor (DAF) and/or CD59/membrane attack complex-inhibitory factor (MACIF). In addition, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) analysis was performed to assess apoptosis of erythroblasts from six patients. On flow cytometric analysis, cases 1 to 6 had positive and negative erythrocyte populations, case 7 intermediate and negative populations, case 8 positive, intermediate, and negative populations, and case 9 a single double-negative population. In addition, cases 1 to 6 and 8 had positive, intermediate, and negative erythroblast populations, while cases 7 and 9 had intermediate and negative populations. The percentage of double-negative erythrocytes showed a significant correlation with that of double-negative erythroblasts (r = .741, P < .05). In seven of nine patients, more erythroblasts than erythrocytes were negative for the two membrane proteins. Also, some patients with an intermediate population of erythrocytes did not necessarily show an increase of PNH II erythroblasts. Apoptosis of PNH erythroblasts was also detected, but the percentage of apoptotic cells in PNH patients showed no difference from that in healthy volunteers. These findings suggest that the final phenotype of mature erythrocytes in PNH is determined during maturation from erythroblasts to erythrocytes by the disappearance or persistence of PNH II erythroblasts. In addition, PNH erythroblasts in vitro may be partly lost by apoptosis, but apoptosis does not play an important role in determining GPI linked protein expression. PMID- 9207482 TI - Bone marrow transplantation in acid sphingomyelinase-deficient mice: engraftment and cell migration into the brain as a function of radiation, age, and phenotype. AB - Types A and B Niemann-Pick disease (NPD) result from the deficient activity of the lysosomal hydrolase, acid sphingomyelinase (ASM). A long-term goal of our research is to evaluate the effects of bone marrow transplantation (BMT) and hematopoietic stem cell gene therapy (HSCGT) on the NPD phenotype. As an initial step toward this goal, we have undertaken a study aimed at optimizing hematopoietic cell engraftment in acid sphingomyelinase "knock-out" (ASMKO) mice. Several parameters were analyzed, including the effects of radiation and donor cell number on survival and engraftment of newborn and adult animals, the number of donor cells detected in the brain posttransplantation, and the levels of ASM activity achieved in the brain. A total of 202 ASMKO and normal animals were transplanted and studied, and the overall conclusions were: (1) newborn ASMKO animals were more susceptible to radiation-induced mortality than normal animals, (2) at low radiation doses, increasing the donor cell number improved engraftment, while this was less evident at the higher radiation doses, (3) engraftment was easier to achieve in normal as compared with ASMKO animals, (4) among newborn transplants, the number of donor cells detected in the brain was directly correlated with engraftment in the blood, (5) more donor cells were detected in the brains of newborn ASMKO animals as opposed to newborn normal animals, and (6) no donor cells were found in the brains of animals transplanted as adults, including those that were highly engrafted in the blood. These results provide important information regarding the design of future BMT and HSCGT studies in ASMKO mice and other mouse models and demonstrate the potential of altering the NPD phenotype by these therapeutic strategies. PMID- 9207483 TI - Granulocyte colony-stimulating factor reduces the capacity of blood mononuclear cells to induce graft-versus-host disease: impact on blood progenitor cell transplantation. AB - The feasibility of transplanting peripheral blood mononuclear cells (PBMC) from granulocyte colony-stimulating factor (G-CSF)-treated normal human donors to myeloablated allogeneic hosts has been demonstrated recently. The current work examined the ability of recombinant G-CSF to alter peripheral blood T-cell function and graft-versus-host disease (GVHD) in a murine model of allogeneic G CSF-mobilized PBMC transplantation. Administration of recombinant G-CSF to C57BL/Ka mice markedly increased the capacity of PBMC to reconstitute lethally irradiated syngeneic hosts. T- and B-lineage lymphocytes were depleted about 10 fold in the bone marrow of the treated mice, and the T-cell yield in the blood was increased about fourfold. The ability of PBMC or purified CD4+ and CD8+ T cells to induce acute lethal GVHD in irradiated BALB/c mice was reduced after the administration of G-CSF. This was associated with decreased secretion of interferon gamma and interleukin-2 (IL-2) and an increased secretion of IL-4. The donor cell inoculum, which was most successful in the rescue of irradiated allogeneic hosts, was the low-density fraction of PBMC from G-CSF-treated mice. These low-density cells were enriched for CD4-CD8-NK1.1+ T cells and secreted about 10-fold more IL-4 than the unfractionated cells from the G-CSF-treated donors. PMID- 9207484 TI - Does the granulocyte-macrophage colony-forming unit content in ex vivo-expanded grafts predict the recovery of the recipient leukocytes? AB - We have investigated the leukocyte-repopulating-predictive value of granulocyte macrophage colony-forming unit (CFU-GM) analyses in ex vivo-expanded versus fresh murine bone marrow (BM) grafts. After the transplantation of graded numbers of normal BM cells (from 15 to 5 x 10(3) CFU-GMs/mice), a dose-dependent increase in the recipient leukocytes was observed between the first and third weeks posttransplantation. During these stages, increases in the graft size of 100-fold improved the leukocyte counts up to 30-fold and shortened the leukopenia period by 5 to 11 days, depending on the leukocyte threshold considered. To investigate whether similar correlations could be established using ex vivo-expanded samples, the size of the CFU-GM population was maximized by means of the preactivation of the BM with 5-fluorouracil (9-day 5FU-BM), followed by 3 days of incubation with interleukin-1 plus stem cell factor. Under these conditions, the CFU-GM content of the ex vivo-expanded grafts was 73-fold higher than that observed in equivalent femoral fractions of normal fresh BM. When equivalent fractions of both graft types were transplanted, an improved leukocyte recovery was observed in mice transfused with the expanded grafts. However, the leukocyte values obtained after the transplantation of the ex vivo-expanded samples were not as high as expected, based on the number of transplanted CFU-GMs. Analyses performed during the second week posttransplantation showed that, in comparison with normal fresh BM, ex vivo-expanded grafts containing 6 to 50 times more CFU-GMs were required to generate a similar number of leukocytes. These results were confirmed in both the peripheral blood leukocytes and the myeloid Gr1+ cells, when similar numbers of CFU-GMs were transfused in the fresh and the ex vivo-expanded BM. The possibility that the preactivation of the ex vivo-expanded grafts with 5FU had a role in this effect was ruled out, because the leukocyte repopulation capacity of fresh 5FU-treated BM was as high as that observed in normal fresh BM which contained a similar number of CFU-GMs. Neither by extending the ex vivo incubation period nor by using other hematopoietic growth factor combinations was the functional capacity of the expanded grafts improved. The results presented in this study are consistent with the belief that ex vivo expansion procedures will be a useful tool for improving the hematologic recovery of patients who receive hematopoietic transplants. However, our data indicate that predicting the leukocyte repopulating capacity of ex vivo-expanded grafts according to correlations established with numbers of fresh CFU-GMs can lead to overestimations of their function, and therefore to unexpected and delayed hematopoietic engraftments. PMID- 9207485 TI - Fludarabine monophosphate reduces the incidence and severity of graft-versus-host disease in a murine model of bone marrow transplantation. PMID- 9207486 TI - The terminology of "Carcinoma cell leukemia"--is an alternative needed? PMID- 9207487 TI - Cutaneous T-cell lymphomas and bacterial superantigens. PMID- 9207488 TI - Nitric oxide modulation of erythropoiesis in rats. PMID- 9207489 TI - Family studies in Scott syndrome. PMID- 9207490 TI - Managed care: emphasis on managed. PMID- 9207491 TI - Merrill C. Sosman Lecture. Surviving managed care. AB - Managed care has arrived, especially on the West Coast. This tsunami is moving eastward at a rapid rate. Perhaps the experience with killer bees will be repeated with managed care, with decreased virulence as this wave moves across the country. Institutions would be wise to prepare for these changes with improved efficiency and cost-effectiveness. Radiologists are particularly at risk and need to aggressively adapt to managed care by becoming informed, developing a clear strategy for coping, and then executing their plan. Evolution will demonstrate once again the survival of the fittest. PMID- 9207493 TI - Malpractice issues in radiology. Informed consent. PMID- 9207494 TI - Machines that learn: can they learn to interpret radiographs? PMID- 9207492 TI - Appropriateness of imaging procedure requests: do radiologists agree? AB - OBJECTIVE: We explored the agreement among radiologists in their evaluation of the appropriateness of individual requests for imaging procedures. MATERIALS AND METHODS: We reviewed 318 noninterventional CT, sonographic, MR imaging, and nuclear medicine procedures ordered at a general internal medicine clinic during 8 months in 1995. Five subspecialty radiologists used data from the radiology request from and clinic notes to independently rate the appropriateness of each requested imaging procedure on a four-point scale. The radiologists were unaware of the results achieved by each procedure. Each case was reviewed by at least three radiologists, of whom at least one had relevant subspecialty expertise. Agreement among radiologists was analyzed using Cohen's kappa statistic and weighted kappa statistics and Cronbach's alpha statistic. RESULTS: Nonchance agreement (kappa) was .19 +/- .05; weighted kappa was .24 +/- .05. Interrater agreement was significantly greater than that expected from chance alone (p < .01). The composite score, defined as the average of the radiologists' scores for each case, showed moderate reliability, as evidenced by a value for Cronbach's alpha of 70. CONCLUSION: In the absence of explicit criteria, we found modest but statistically significant agreement among radiologists about the appropriateness of individual requests for imaging procedures. The disagreement among radiologists highlights the importance of developing well-reasoned, explicit criteria by which to judge the appropriateness of diagnostic radiology procedures. Further study is needed to elucidate the relationship between appropriateness and actual patient outcomes. PMID- 9207495 TI - Wavelet transform-based image compression for transmission of MR data. AB - OBJECTIVE: The purpose of this study was to develop an effective, inexpensive teleradiology image-transmission system for transfer of MR studies using wavelet transform image compression. CONCLUSION: We describe an efficient system for implementing teleradiology capability for transmission of diagnostic MR images. The system uses the wavelet transform to achieve greater than 90% image compression, with standard modern transmission times of less than 5 sec per compressed image. The method is inexpensive and can be implemented using commonly available workstation and MR scanner capabilities. PMID- 9207496 TI - Voice recognition in radiology reporting. AB - OBJECTIVE: Recent advances in computer hardware and software technology have improved voice recognition systems used for radiology reporting. We describe and analyze this technology and some of its costs as well as the prospects for using voice recognition systems in the transcription of radiology reports. Factors to be considered in choosing a system include language model, speech flow, vocabulary size, and methods of acoustic modeling. CONCLUSION: Careful evaluation of individual radiology practice will permit proper introduction of voice recognition technology into mainstream clinical use. With this new technology, radiologists can use normal speech patterns to dictate while reviewing films. Also, this technology can use standard personal computers to reduce hardware costs to a level acceptable to radiology departments. PMID- 9207497 TI - Antibiotic prophylaxis in vascular and interventional radiology. PMID- 9207498 TI - Evaluation of renal artery stenosis with dynamic gadolinium-enhanced MR angiography. AB - OBJECTIVE: The purpose of this study was to compare dynamic gadolinium-enhanced three-dimensional spoiled gradient-recalled MR angiography with conventional arteriography in the evaluation of proximal renal artery stenosis (RAS). MATERIALS AND METHODS: MR angiography and conventional arteriographic examinations of 30 patients evaluated for RAS were analyzed retrospectively. Three-dimensional MR angiography was performed with an RF spoiled gradient recalled imaging sequence acquired during the dynamic i.v. injection of gadolinium (0.2-0.3 mmol/kg), MR data and conventional arteriograms were independently evaluated for the number and location of renal arteries and the degree and location of stenoses. The patients had a mean age of 70 years old and a mean serum creatinine level of 2.9 mg/dl, reflecting a population in whom atherosclerotic RAS was the primary concern. RESULTS: Gadolinium-enhanced MR angiography revealed 100% of main renal arteries. For RAS of 50% or greater occlusion, the technique was 100% sensitive and 71% specific; the negative predictive value was 100%. The technique was 100% sensitive and 71% specific for RAS of 75% or greater occlusion. CONCLUSION: Dynamic gadolinium-enhanced three dimensional spoiled gradient-recalled MR angiography has a high sensitivity for revealing proximal RAS and is a quick and reliable technique for obtaining helpful anatomic information. PMID- 9207499 TI - MR angiography of the iliac and upper femoral arteries using four different inflow techniques. AB - OBJECTIVE: The purpose of this study was to compare two inflow MR angiography pulse sequences obtained with and without systolic synchronization. We also compared these two MR angiography pulse sequences with conventional angiography. SUBJECTS AND METHODS: Thirty-one consecutive patients who were scheduled for conventional angiography because of symptomatic atherosclerotic occlusive disease of the iliac or femoral artery underwent MR angiography using four different MR angiography techniques. These techniques consisted of a multiple two-dimensional inversion prepulse gradient-recalled echo technique (turbo field-echo) obtained with and without systolic synchronization and a multiple two-dimensional gradient recalled echo technique (fast field-echo) obtained with and without systolic synchronization. We then compared image quality and our ability to detect and grade degree and length of stenosis, using conventional angiography as the gold standard. RESULTS: The systolic-synchronized turbo field-echo sequence produced the best results both objectively and subjectively. Comparing systolic synchronized turbo field-echo and fast field-echo techniques with conventional angiography regarding detection and grading degree of stenoses, we found no statistically significant differences. CONCLUSION: Systolic synchronization proved to be of significant importance for image quality. The systolic synchronized turbo field-echo pulse sequence proved to be superior to the other three MR angiography techniques. PMID- 9207500 TI - Rapid revascularization of an embolic superior mesenteric artery occlusion using pulse-spray pharmacomechanical thrombolysis with urokinase. PMID- 9207501 TI - Aortocaval fistula: diagnosis with MR angiography. PMID- 9207503 TI - Radiologic issues in lung transplantation for end-stage pulmonary disease. AB - Lung transplantation has evolved into an effective therapy for end-stage pulmonary disease. The radiologist has an important role in the management of these patients before and after transplantation. Unfortunately, the radiologic findings of the major complications (i.e., reperfusion edema, infection, rejection) that occur after transplantation overlap and are often nonspecific. Clinical correlation, bronchoalveolar lavage, and transbronchial biopsy are usually required for accurate differentiation. The following rules of thumb may be helpful: Radiographic opacities seen during the first week are usually due to reperfusion edema: persistent or progressive opacities beyond the first week suggest infection or acute rejection: infection in the first month is usually bacterial, and opportunistic pneumonia is more common thereafter, nodular opacities are usually due to infection or posttransplantation lymphoproliferative disorders but can also be due to transbronchial lung biopsy; and progressive bronchial dilatation and air trapping seen on expiratory CT are useful signs of BOS. PMID- 9207502 TI - Human gene therapy for melanoma: CT-guided interstitial injection. AB - OBJECTIVE: Our intent is to describe the role of CT in the intratumoral injection of Allovectin-7 (Vical, San Diego, CA), an allogeneic class I major histocompatibility complex antigen, HLA-B7, formulated with cationic lipid, in the treatment of metastatic malignant melanoma. MATERIALS AND METHODS: Ten patients with metastatic malignant melanoma were treated with gene therapy in which we used CT-guided intratumoral injection of plasmid DNA containing the HLA B7 gene. This therapy was part of a phase I gene therapy trial in patients with metastatic melanoma. CT guidance was chosen as an accurate way to direct gene delivery in patients with deep, impalpable lesions. Tumor locations included pulmonary, mediastinal, hepatic, adrenal, and paracaval sites. Patients in the CT protocol underwent baseline CT studies. Examinations were repeated 2, 4, and 8 weeks after gene therapy and thereafter at 3-month intervals. Both injected and noninjected tumors were measured. CT-guided injections of 10, 50, or 250 micrograms of plasmid DNA were performed with 22-gauge spinal needles. Injection volumes were between 1.0 and 4.0 ml, depending on tumor size. CT-guided core biopsy specimens were obtained (with 18- or 20-gauge needles) from the selected tumor before therapy and 2, 4, and 8 weeks after therapy to assess HLA-B7 plasmid DNA and gene expression. Peripheral blood was analyzed for cytotoxic T lymphocytes directed against HLA-B7. RESULTS: CT-guided intratumoral injections were successful in delivering genetic material to all patients with impalpable tumors. Significant responses (as defined by a decrease of 25% or more in the product of the length and width of the injected tumor) were observed in six of the 10 patients. One of these six patients who had a solitary lesion remains free of disease 19 months after gene therapy. HLA-B7 protein expression was detected in 89% of biopsy specimens, and plasmid DNA and messenger RNA were detected in 56% and 22% of biopsy specimens, respectively. CONCLUSION: CT provides a safe, accurate, and efficacious way to monitor and assess tumor progression and response, and it provides guidance for biopsies and intratumoral injections during gene therapy. Significant responses in injected tumors of six of the 10 patients in our study suggest that further clinical trials of this gene therapy are warranted. PMID- 9207504 TI - Mosaic pattern of lung attenuation on CT scans: frequency among patients with pulmonary artery hypertension of different causes. AB - OBJECTIVE: The purpose of this study was to determine the frequency with which a mosaic pattern of lung attenuation is seen on chest CT scans in patients with various causes of pulmonary artery hypertension (PAH). MATERIALS AND METHODS: Chest CT scans of 64 patients with known PAH were reviewed to assess the patterns of lung attenuation. Patterns of lung attenuation were divided into three categories: class I, homogeneous lung parenchymal attenuation; class II, slightly heterogeneous lung attenuation that does not conform to the anatomic boundaries of the secondary pulmonary lobule; and class III (mosaic pattern), heterogeneous lung attenuation in geographic regions with well-defined borders corresponding to the anatomic units of single or multiple secondary pulmonary lobules. The patients medical histories were reviewed to determine the primary cause of PAH for each patient. RESULTS: Peak pulmonary artery pressure of the patients in our study averaged 74 mm Hg (range, 36-194 mm Hg). Twenty-one patients had PAH due to lung disease: 17 patients, due to cardiac disease; and 23 patients, due to vascular disease. Three other patients had PAH due to miscellaneous causes. Of the 23 patients with PAH due to vascular disease, 17 patients (74%) had a mosaic pattern of lung attenuation. Of the 21 patients with PAH due to lung disease, one patient (5%) had a mosaic pattern of lung attenuation. Among the 17 patients with PAH due to cardiac disease, two patients (12%) had a mosaic pattern of lung attenuation. A mosaic pattern of lung attenuation was seen significantly more often in patients with PAH due to vascular disease than in patients with PAH due to cardiac or lung disease. CONCLUSION: A mosaic pattern of lung attenuation can be seen on CT scans in patients with PAH due to vascular disease, cardiac disease, or lung disease. However, the mosaic pattern is seen significantly more often in patients with PAH due to vascular disease than in patients with PAH due to cardiac or lung disease. PMID- 9207505 TI - Pulmonary involvement in polymyositis and dermatomyositis: sequential evaluation with CT. AB - OBJECTIVE: The characteristic findings of pulmonary involvement in polymyositis (PM) or dermatomyositis (DM) and the change in findings before and after treatment were evaluated with sequential high-resolution CT studies. MATERIALS AND METHODS: CT images of pulmonary involvement in 19 patients with PM or DM were reviewed. During a period of 2-61 months, 17 of these patients underwent sequential CT before and after treatment with corticosteroids, immunosuppressants, or both. RESULTS: Findings of the initial CT studies included pleural irregularities and prominent interlobular septa (n = 19), ground-glass attenuation (n = 19), patchy consolidation (n = 19), parenchymal bands (n = 15), irregular peribronchovascular thickening (n = 15), and subpleural lines (n = 7). Honeycombing was not detected on any CT images. These findings were more remarkable in the lower portion and the subpleural area of the lungs. In 16 of the 17 patients who underwent sequential CT conditions such as patchy consolidation, parenchymal bands, and irregular peribronchovascular thickening improved, becoming pleural irregularities and prominent interlobular septa, ground-glass attenuation, and subpleural lines on follow-up CT scans. CONCLUSION: Consolidation with patchy and subpleural distribution, parenchymal bands, and irregular peribronchovascular thickening were characteristic and reversible CT findings in patients with PM or DM. PMID- 9207506 TI - CT appearance of the pleural space after talc pleurodesis. AB - OBJECTIVE: This study describes the CT appearance of the pleural space after talc pleurodesis. CONCLUSION: After talc pleurodesis, the pleural space reveals variable degrees of pleural thickening and nodularity, often with a residual loculated effusion. High-attenuation areas representing talc deposits are expected and should not be confused with other pleural abnormalities. PMID- 9207507 TI - Evaluation of electron beam CT coronary angiography in healthy subjects. AB - OBJECTIVE: This study evaluated the performance characteristics of electrocardiographically triggered, contrast-enhanced electron beam CT (EBCT) in defining the coronary artery lumen in healthy subjects. SUBJECTS AND METHODS: The coronary arteries of 11 healthy young men (mean age, 24 years old) were evaluated by contrast-enhanced EBCT. Measured parameters included degree of luminal enhancement, intravascular contrast-to-noise ratio, apparent luminal diameter, and length of continuously visualized lumen (100-H threshold for diameter and length measurements). RESULTS: Aortic blood pool attenuation was 44 +/- 5 H (mean +/- SD) before and 278 +/- 35 H after IV injection of contrast material. Contrast to-noise ratios ranged from a high of 10.0 +/- 2.6 in the proximal right coronary artery to a low of 3.2 +/- 2.7 in the distal left circumflex artery, decreasing from proximal to distal within each vessel. Apparent luminal diameters were as follows: left main coronary artery, 4.5 +/- 0.6 mm; left anterior descending artery, 3.7 +/- 0.5 mm; left circumflex artery, 2.9 +/- 0.6 mm; and right coronary artery, 3.5 +/- 0.5 mm. The mean lengths of visualized lumina were as follows: left main coronary artery, 10 +/- 4 mm; left anterior descending artery, 65 +/- 26 mm; left circumflex artery, 45 +/- 20 mm; and right coronary artery, 58 +/- 24 mm. CONCLUSION: EBCT angiography can reveal the lumen of long segments of the major coronary arteries. PMID- 9207508 TI - Estimation of the left ventricular ejection fraction using a novel multiphase, dark-blood, breath-hold MR imaging technique. AB - OBJECTIVE: In this paper, we evaluate a recently proposed dual-phase dark-blood MR sequence for estimating the left ventricular ejection fraction, compare Simpson's method of estimation of ejection fraction with a model based on the biplane method, assess the reproducibility of both methods, and finally, test a semiautomated method for contouring the endocardial border. SUBJECTS AND METHODS: An MR pulse sequence was implemented to acquire cardiac images in both diastolic and systolic phases within a single breath-hold. A special magnetization preparation scheme rendered blood dark while a segmented acquisition allowed breath-hold scan times. Five healthy volunteers and five patients with cardiac disease were imaged. Ejection fractions were estimated using (1) long-axis and four-chamber biplane views with an ellipsoid model and (2) a series of short-axis views in combination with Simpson's model. These values of ejection fractions were then compared with values obtained from echocardiography. RESULTS: Estimates of ejection fractions obtained using biplane ellipsoid volume and Simpson's rule methods varied by 14% in healthy volunteers. However, for patients with severe cardiomyopathy, differences between the values of ejection fraction obtained with the two methods varied by as much as 150%. Ejection fraction estimates obtained from MR images with the biplane ellipsoid method and from echocardiography varied by approximately 14% for all subjects. Ejection fraction estimates obtained with the semiautomated algorithm agreed well with estimates obtained with manual contours made by experienced radiologists. Intraobserver variability was low for both the short-axis (3%) and biplane (4%) methods. However, interobserver variability of the biplane method (12%) exceeded that of the short-axis method (4%). Interexamination variability (9%) was the largest factor in determining the reproducibility of the ejection fraction estimates. CONCLUSION: Breath-hold dark blood MR imaging technique with simultaneous acquisition of a series of short axis views during systolic and diastolic phases permits rapid and accurate estimates of ejection fractions in healthy subjects and in patients. Model based biplane MR imaging methods are less reliable in patients with global cardiomyopathy. The estimation of ejection fractions can be automated using the proposed contouring algorithm and the dark-blood short-axis views. PMID- 9207509 TI - A focused appendiceal CT technique to reduce the cost of caring for patients with clinically suspected appendicitis. AB - OBJECTIVE: This investigation analyzed the potential impact on hospitalwide variable costs and total costs of introducing a focused helical CT technique for diagnosing appendicitis. MATERIALS AND METHODS: This investigation had three components. First, we retrospectively reviewed the records of 651 patients who had admitting diagnoses of appendicitis. Second, we determined variable costs and total costs for the components of these patients' care. Third, we projected change in costs if the focused appendiceal CT technique had been used to guide patient management. RESULTS: Using focused appendiceal CT in 100 patients would have decreased the number of nontherapeutic appendectomies by 13. The average length of stay for observation would have also decreased by 1 day because the diagnosis of an abnormal appendix or an alternative diagnosis would have been made more quickly. Savings in variable costs and total costs would have been $23,030 and $45,556 per 100 patients, respectively. CONCLUSION: Routine use of focused appendiceal CT would lower the costs of caring for patients with clinically suspected appendicitis. PMID- 9207510 TI - Arterial versus portal venous helical CT for revealing pancreatic adenocarcinoma: conspicuity of tumor and critical vascular anatomy. AB - OBJECTIVE: The purpose of this study was to determine the conspicuity of pancreatic adenocarcinoma and surrounding critical pancreatic vascular structures on helical CT scans obtained during the arterial and portal venous phases of enhancement. SUBJECTS AND METHODS: Forty patients with pancreatic adenocarcinomas under-went dual-phase helical CT (3-mm collimation; l-mm overlapping reconstructions; 160 ml contrast medium injected at 4 ml/sec; scan delay: 18 sec for arterial phase, 60 sec for portal venous phase). Tissue enhancement and differences in tumor-to-pancreas contrast were compared. Quantitative evaluation was also done for the aorta, the superior mesenteric artery and vein, and the splenic and portal veins. RESULTS: Tumor conspicuity was significantly greater in the portal venous phase, when the tumor-to-pancreas contrast difference was 54 +/ 31 H, than in the arterial phase, when the difference was 31 +/- 29 H. Enhancement values of critical pancreatic venous structures were significantly greater in the portal venous phase than in the arterial phase. CONCLUSION: Arterial-phase helical CT in patients with pancreatic adenocarcinoma is of limited benefit: lesion conspicuity is suboptimal and depiction of venous anatomy is inferior to the depiction possible with venous-phase helical CT. PMID- 9207511 TI - Hepatic lesion characterization in cirrhosis: significance of arterial hypervascularity on dual-phase helical CT. AB - OBJECTIVE: Our goal was to evaluate the diagnostic significance of the presence and pattern of arterial hypervascularity in lesions detected on dual-phase helical CT in cirrhotic patients. MATERIALS AND METHODS: Fifty-eight lesions greater than 1 cm in size were prospectively identified in 26 patients with end stage liver disease who had undergone dual-phase helical CT for preoperative liver transplantation evaluation. All 26 patients had diagnoses proven by histologic evaluation or by clinical criteria. All arterial phase scans were retrospectively reviewed and lesions were categorized for the presence and pattern of arterial hypervascularity. Radiologic findings were correlated with histopathologic data. RESULTS: Thirty-seven of the 58 lesions had hypervascular components on arterial phase scans. All 37 of these lesions were found to represent hepatocellular carcinoma (HCC) (positive predictive value, 100%). Of the 21 remaining hypovascular lesions, 17 were HCC and four were benign (positive predictive value, 81%). Of the nine patients in whom all lesions were hypovascular, six had HCC (positive predictive value, 66%). The value of the presence of arterial hypervascularity for diagnosing HCC was statistically significant (p < .05). However, the presence or absence of arterial hypervascularity and the specific enhancement pattern revealed by helical CT did not correlate with histologic grading. CONCLUSION: The presence of hypervascularity in hepatic masses found in cirrhotic patients is highly predictive of malignancy. PMID- 9207512 TI - Helical CT during arterial portography and hepatic arteriography for detecting hypervascular hepatocellular carcinoma. AB - OBJECTIVE: We assessed whether helical CT hepatic arteriography (CTHA) improves detection of hypervascular hepatocellular carcinoma (HCC) when used in conjunction with helical CT obtained during arterial portography (CTAP). SUBJECTS AND METHODS: One hundred two patients with 254 hypervascular HCC nodules underwent both CTHA and CTAP. CTHA and CTAP were separately interpreted for detection of hypervascular HCC nodules by three observers who were unaware of tumor burden in the liver, lodized oil CT performed 1 week and 1 month after transcatheter arterial chemoembolization with iodized oil was the gold standard. Sensitivity and positive predictive value for CTHA alone, CTAP alone, and CTAP and CTHA combined were calculated and compared using the McNemar test. RESULTS: We found no significant difference between the sensitivity of CTAP (85%) and CTHA (87%) for revealing hypervascular HCC nodules. The combination of CTAP and CTHA showed significantly higher sensitivity than either CTAP or CTHA alone for revealing HCC nodules smaller than 20 mm in diameter (p < .01). CONCLUSION: The combination of CTHA and CTAP is recommended to improve the detection sensitivity of small hypervascular HCC nodules. PMID- 9207513 TI - Cavernous hemangiomas of the liver: enlargement over time. AB - OBJECTIVE: It has been reported that cavernous hemangiomas of the liver either do not enlarge over time, or, of the very few lesions that have shown an increase in diameter on follow-up imaging studies, the increase has been minimal. The objective of this paper is to report growth of four cavernous hemangiomas that were shown to have doubled to tripled in diameter on follow-up imaging studies done between 34 months and 10.5 years after the initial diagnosis. CONCLUSION: This report indicates that hepatic hemangiomas can grow significantly in diameter, although such growth is unusual. Despite the growth of the lesion, however, if the imaging features are characteristic of hemangioma an imaging diagnosis can still be made confidently. PMID- 9207514 TI - Flow pulsatility in the portal venous system: a study of Doppler sonography in healthy adults. AB - OBJECTIVE: The purpose of our study was to describe Doppler sonography patterns of venous flow in the portal system of healthy subjects and to compare pulsatility of flow with subjects' body mass, degree of inspiration, and body position. SUBJECTS AND METHODS: Doppler signals from the main, right, and left portal veins; superior mesenteric vein; splenic vein; and inferior vena cava of 23 healthy adults were prospectively studied. Pulsatility of flow was quantified using an index of venous pulsatility (VPI = [maximum frequency shift-minimum frequency shift]/maximum frequency shift). Antegrade flow peak velocities were also related to ECG tracings the time between two R waves being divided into four equal parts for analysis. The caliber variations of the main portal vein and inferior vena cava were measured with M-mode sonography. Doppler tracings were obtained with subjects in supine and sitting positions and during mid and deep inspiration. The subjects' heights and weights were obtained and the body mass index calculated (weight/[height2]). RESULTS: In the portal vein, the VPI was 0.48 +/- 0.31 (mean +/- SD). Marked pulsatility of venous flow (VPI > 0.5) was found in 12 of 23 subjects. We found an inverse correlation between the VPI and the subjects' body mass index (r = -.76; p < .001). Portal vein pulsatility decreased significantly during sitting (p < .05) and deep inspiration (p < .01). The portal VPI was correlated with caliber variation of the inferior vena cava (r = .59; p < .05). In the portal venous system, antegrade flow peak velocities occurred most often during the third quarter of the cardiac cycle, particularly in the splenic vein. CONCLUSION: Doppler sonography shows pulsatile portal venous flow in healthy adults, especially in thin subjects. This pulsatility has an inverse correlation to body mass. The finding of a pulsatile portal vein needs to be interpreted in clinical context and does not necessarily imply dysfunction of the right side of the heart. PMID- 9207515 TI - The role of transvaginal sonography and endometrial biopsy in the evaluation of peri- and postmenopausal bleeding. AB - OBJECTIVE: The number of women seeking medical attention for peri- and postmenopausal bleeding (PMB) has been increasing. Determining the cause of PMB is essential in planning appropriate therapy. In these women, transvaginal sonography (TVS) is a sensitive means for diagnosing the causes of such bleeding, yet endometrial biopsy (EMB) is still preferred as the first diagnostic test. We prospectively compared TVS with aspiration biopsies of the endometrium in the examination of women with PMB. SUBJECTS AND METHODS: Between mid April 1994 and December 1995, 329 consecutive perimenopausal women underwent EMB. Of these EMBs 302 had negative results. We prospectively obtained TVS in 259 of these 302 women within 1 month of EMB (range, 10 days to 2 months) when the results of biopsy were negative. Forty-three patients were lost to follow-up. In 59 women who had endometrial thickening greater than 5 mm, dilatation and curettage, hysteroscopy, or hysterectomy was performed. Ninety-four of the 130 women who were found at TVS to have fibromyomata or diffusely enlarged uteri underwent hysterectomy for pathologic confirmation. The remaining 36 women with fibromyomata or diffusely enlarged uteri had no pathologic confirmation of their TVS findings. Twenty-one of 64 women with endometria thinner than 5 mm underwent dilatation and curettage, and 43 of these women were lost to follow-up. RESULTS: In 259 patients who underwent TVS, 57 patients who had an endometrium thicker than 5 mm and an endoluminal mass on hysterosonography had false-negative results on aspiration biopsies. Of the 18 patients who had malignancies in this series, 12 had false negative results on biopsies. In the 94 patients with an enlarged uterus and negative EMB results who underwent hysterectomy, we found 87 with fibroids, three with adenomyosis, and four with sarcomas. Of the 64 women with endometria thinner than 5 mm seen on TVS, 21 had negative results from dilatation and curettage. CONCLUSION: EMB alone is not sufficient for screening women for PMB. TVS appears to be more sensitive than is EMB for the detection of abnormalities, particularly those outside the endometrium. For these reasons, TVS should be the initial screening test when examining women with PMB. PMID- 9207516 TI - Percutaneous nephrostomy: placement under CT and fluoroscopy guidance. AB - OBJECTIVE: The purpose of this paper is to present our experience with CT- and fluoroscopy-guided percutaneous nephrostomy tube (PNT) placement and to describe the technique of placement with patients in the supine-oblique position. MATERIALS AND METHODS: A CT scanner was coupled with a ceiling-mounted C-arm for fluoroscopy at the CT table, PNT was done with CT localization and fluoroscopic control. We attempted 148 placements in 129 patients, with most patients in the supine or the supine-oblique position. RESULTS: Successful PNT placement was achieved in 144 (97%) of 148 kidneys. In the native kidney group, 101 (81%) of 124 PNTs were placed with the patients in the supine-oblique position. We experienced no major complications, three (2%) minor complications, and two (1%) catheter dislodgments. CONCLUSION: CT-fluoroscopy control allows routine PNT placement with patients in the supine or the supine-oblique position and is associated with the lowest complication rate to our knowledge. PMID- 9207517 TI - CT appearance of transitional cell carcinoma of the renal pelvis: Part 1. Early stage disease. PMID- 9207518 TI - CT appearance of transitional cell carcinoma of the renal pelvis: Part 2. Advanced-stage disease. PMID- 9207519 TI - Primary adrenal ganglioneuroma: CT findings in four patients. AB - OBJECTIVE: The purpose of this study was to characterize the appearance of adrenal ganglioneuroma on CT scans and to define the specific imaging features of the tumor. CONCLUSION: Adrenal ganglioneuroma is an uncommon benign tumor that is revealed as a solid adrenal mass on CT scans. Specific CT features such as calcification and lesion enhancement may simulate a primary adrenal carcinoma. However, in our four patients, no signs of local invasion or vascular extension were noted. The specific diagnosis of adrenal ganglioneuroma requires either biopsy or surgical removal for documentation. PMID- 9207520 TI - Continent urinary diversion procedures: radiographic appearances and potential complications. PMID- 9207521 TI - CT appearance of parapneumonic effusions in children: findings are not specific for empyema. AB - OBJECTIVE: Although definitive differentiation of empyema from transudative parapneumonic effusion is based on the analysis of pleural fluid, certain CT findings have been described as highly suggestive of empyema. This study compares the CT findings of parapneumonic effusions with the results of thoracentesis, thoracoscopy, or both to determine whether these CT findings can reliably differentiate empyemas from transudative parapneumonic effusions in children. MATERIALS AND METHODS: CT scans obtained to evaluate pleural or parenchymal complications of pneumonia were reviewed. Parapneumonic effusions were evaluated for the CT findings of pleural enhancement; parietal pleural thickening; thickening, increased attenuation, or both of the extrapleural subcostal fat: and edema of the extracostal chest wall. Each parapneumonic effusion was assigned a CT score on the basis of these CT findings. Individual CT findings and the CT score were correlated with the presence of empyema as determined by thoracentesis or thoracoscopy. RESULTS: Thirty patients were identified as having a parapneumonic pleural effusion revealed on contrast-enhanced CT scans and by pleural fluid analysis. Twenty-one of these parapneumonic effusions met the clinical criteria for empyema, and nine were considered not to be empyemas. Neither any individual CT finding nor the CT score accurately differentiated empyema from transudative parapneumonic effusions (p > .1): pleural enhancement (empyema 100%, transudative effusion 89%), pleural thickening (empyema 57%, transudative effusion 56%), abnormal extrapleural space (empyema 66%, transudative effusion 67%), extracostal chest wall edema (empyema 33%, transudative effusion 56%), and average CT score (empyema 2.5. transudative effusion 2.3). CONCLUSION: CT characteristics of parapneumonic effusions do not allow radiologists to accurately predict empyema. The presence or absence of such CT findings should not influence therapeutic decisions concerning the management of parapneumonic effusions. PMID- 9207522 TI - Normal gadolinium-enhanced MR images of the developing appendicular skeleton: Part I. Cartilaginous epiphysis and physis. AB - OBJECTIVE: We have used gadolinium-enhanced MR imaging to define the expected normal appearance of the developing cartilaginous epiphyses and physes in neonates, infants, and children and to define the changes with maturity in epiphyseal vascular pattern. MATERIALS AND METHODS: We analyzed gadolinium enhanced MR images of 80 normal epiphyses in 48 neonates, infants, and children who were 1 month to 15.5 years old. We studied the differences in enhancement ratios for the epiphyses and physes and the epiphyseal vascular pattern at various development stages. We correlated the MR imaging findings with histologic and injection studies of immature epiphyses. RESULTS: Gadolinium enhancement allowed differentiation between physeal and epiphyseal cartilage and revealed epiphyseal vascular canals. Enhancement proved to be greater in the physeal than in the epiphyseal cartilage (p < .001). In the unossified epiphysis, the vascular canals were mainly parallel. After the development of the secondary ossification center, these canals came to have a radial pattern (p < .0001). Comparison with cadaveric specimens confirmed how, with age, the arrangement of these canals changed. Also, physeal enhancement decreased with physeal closure. CONCLUSION: Gadolinium-enhanced MR imaging reveals differential enhancement of the physis, epiphyseal vascular canals, and epiphyseal cartilage. The pattern of epiphyseal vessels and degree of enhancement of the physis change with maturity. PMID- 9207523 TI - Normal gadolinium-enhanced MR images of the developing appendicular skeleton: Part 2. Epiphyseal and metaphyseal marrow. AB - OBJECTIVE: We have studied how gadolinium enhancement of T1-weighted MR images affects the expected normal differences in signal intensity between metaphyseal hematopoietic and epiphyseal fatty marrow. We have also analyzed how enhancement affects the expected normal changes in the MR images of the marrow due to fatty conversion. MATERIALS AND METHODS: We analyzed gadolinium-enhanced MR images of normal distal femurs in 18 immature rabbits that were 5-11 weeks old and of normal proximal femurs in 18 infants, children, and young adults who were 2 months to 21 years old. In all subjects, we studied the change with age in signal intensity and enhancement ratio of the epiphyseal and metaphyseal marrow. In the rabbits, marrow composition and transformation were histologically verified. RESULTS: On unenhanced T1-weighted MR images of the rabbits and of the infants, children, and young adults, epiphyseal signal intensity always exceeded metaphyseal signal intensity; however, the enhancement ratio was always greater in the metaphysis. The signal intensity in metaphyseal and epiphyseal marrow on unenhanced MR images increased with age. However, enhancement ratios decreased with age in both areas. In the rabbits, histologic studies showed more fatty marrow in the epiphysis than in the corresponding metaphysis and an age-related increase in marrow fat at both sites. CONCLUSION: In the marrow of the extremities, gadolinium enhancement is greater in the (hematopoietic) metaphysis than in the (fatty) epiphysis. In both areas, enhancement decreases as the marrow becomes more fatty. On T1-weighted images, administration of a gadolinium containing contrast agent reduces the normal contrast between hematopoietic and fatty marrow and obscures the changes in marrow signal intensity due to fatty conversion. PMID- 9207524 TI - Genitourinary complications of Crohn's disease in pediatric patients. AB - OBJECTIVE: The purpose of this study was to determine the incidence and types of genitourinary complications in pediatric patients with Crohn's disease. CONCLUSION: Genitourinary complications of Crohn's disease in children are not rare and may be the initial presentation of the condition. PMID- 9207525 TI - High-resolution MR imaging of the anal sphincter in children: a pilot study using endoanal receiver coils. AB - OBJECTIVE: The purpose of this study was to obtain high-resolution MR images of the various components of the anal sphincter complex in children who have anorectal disorders. We therefore used dedicated endoanal receiver coils for MR imaging. CONCLUSION: Our pilot study suggested that MR imaging that uses a dedicated endoanal coil may have considerable diagnostic potential in children who have anorectal disorders. PMID- 9207526 TI - Spinal level of fetal myelomeningocele: does it influence ventricular size? AB - OBJECTIVE: Our objective was to determine whether the level of the spinal defect influences ventricular size in fetuses with myelomeningoceles. MATERIALS AND METHODS: Sonograms of 51 fetuses with open spina bifida were reviewed to determine the gestational age, ventricular atrial diameter, severity of posterior fossa deformity, and the level of the spinal defect. Four categories for spinal defect level were used: sacral, low lumbar, high lumbar, and thoracic. Regression models for ventricular atrial diameter were fit, adjusting for gestational age, posterior fossa deformity, and spinal defect level. A Fisher's exact test was used to investigate a relationship between the level of the spinal defect and the severity of the posterior fossa deformity. RESULTS: Spinal defect level was distributed among the four categories as follows: sacral (n = 7), low lumbar (n = 30), high lumbar (n = 10), and thoracic (n = 4). The level of the spinal defect did not significantly affect ventricular size (p > .1), and the level of the spinal defect did not show any relationship to the severity of the posterior fossa deformity (p > .8). CONCLUSION: The level of the spinal defect does not independently affect the degree of ventriculomegaly or severity of the posterior fossa deformity in fetuses with myelomeningoceles. PMID- 9207527 TI - Kinematic CT of the patellofemoral joint. AB - OBJECTIVE: The purpose of this study was to determine if kinematic CT can be applied to the patellofemoral joint using current slip-ring CT scanner design in patients with anterior knee pain and thus a suspected patellar tracking disorder. SUBJECTS AND METHODS: Twenty knees in 18 patients with anterior knee pain were evaluated with kinematic CT. A single 10-sec exposure of the patellofemoral joint was obtained during active flexion and extension. Static nonkinematic and loaded kinematic examinations were compared with unloaded kinematic studies in a subset of patients. The changes in lateral patellofemoral angle and lateral shift were measured. Video cine viewing of patellofemoral motion was used to subjectively grade image quality and patellofemoral abnormalities by consensus. RESULTS: Kinematic CT was successfully used in all 20 knees. In nine knees studied with static nonkinematic and unloaded kinematic images, the lateral patellofemoral angle improved an average of 4" on the kinematic images. In addition, lateral shift improved by an average of 3%, an improvement that was statistically significant (p = .01). In 10 knees studied with and without loading, the lateral patellofemoral angle decreased an average of 3% with loading. No significant change was seen in lateral shift. In all patients, cine viewing was thought to be more useful than single images. Cine viewing was of good or diagnostic quality in all 20 knees studied. Lateral patellar translation during extension was detected in eight of 20 kinematic studies. Lateral patellar tilting also was detected in eight of 20 kinematic studies. Narrowing of the articular space was detected in 12 of 20 knees. Six knees were determined to be normal. CONCLUSION: Kinematic CT with slip-ring technology is a new technique that can be easily performed on the patellofemoral joint. This technique shows promise as a tool for determining the cause of anterior knee pain. PMID- 9207528 TI - MR imaging findings in anterior cruciate ligament reconstruction: evaluation of notchplasty. AB - OBJECTIVE: Notchplasty, resection of bone from the roof and lateral side of the inter-condylar notch, is frequently performed during anterior cruciate ligament (ACL) reconstruction to avoid abrasion or deflection of the ACL graft by the femur (graft impingement). Graft impingement can develop months after the initial reconstruction despite adequate notchplasty. Such impingement has recently been attributed to fibrocartilage overgrowth of the notchplasty site. The objectives of this study were to evaluate the MR appearance of the notchplasty site immediately after surgery and 6 months later and to investigate whether cartilage overgrowth of the notchplasty site can be detected with MR imaging. SUBJECTS AND METHODS: Thirty-three patients who underwent bone-tendon-bone autograft ACL reconstruction had MR scans of the intercondylar notch 6 months after surgery. Twenty-five of these patients also had MR scans in the immediate postoperative period. The appearance of the notchplasty site and evidence for cartilage overgrowth of the notchplasty site were evaluated. Arthroscopic correlation was available for six patients. RESULTS: On MR images, the margins of the notchplasty site were identified in all patients. Contour of the notchplasty site along the lateral notch wall was frequently concave toward the notch, a finding not commonly seen in an unoperated notch. Six months later, the shape of the notchplasty site had not changed, but evidence for recortication of the notchplasty site (seen as a layer of signal void 0.5-1.5 mm in thickness over the previously exposed cancellous bone) was seen in 94% of the patients. Additionally, a second layer of signal void, 1-5 mm in thickness with signal intensity identical to that of hyaline cartilage, was seen overlying the notchplasty site in 64% of patients. Arthroscopic correlation available in six patients suggested that this second layer represented fibrocartilage overgrowth of the notchplasty site. CONCLUSION: Characteristic changes of the intercondylar notch after notchplasty can be seen on MR images. This preliminary study also suggests that a thin layer of cortical bone forms over the notchplasty site in most patients within 6 months of surgery. Perhaps more significantly, overgrowth of the notchplasty site by fibrocartilage may also be detected in some patients. Further experience is needed to determine whether MR imaging is a useful method for identifying cartilage overgrowth after notchplasty. PMID- 9207529 TI - MR imaging of anterior cruciate ligament reconstruction. PMID- 9207530 TI - Palpation-directed (non-fluoroscopically guided) saline-enhanced MR arthrography of the shoulder. PMID- 9207531 TI - MR imaging of lumbar spondylolysis: the importance of ancillary observations. AB - OBJECTIVE: The purpose of this study was to determine the frequency of characteristic ancillary MR findings in patients with lumbar spondylolysis. MATERIALS AND METHODS: The radiology reports and clinical records of 64 patients (16 female, 48 male; 12-77 years old) with 66 levels of lumbar spondylolysis who had undergone MR imaging were retrospectively reviewed. Spondylolysis was established by conventional radiography in all 64 patients and by CT in 18 patients. The proportion of patients with spondylolysis in whom sagittal MR images showed ancillary findings of an increased sagittal diameter of the spinal canal, reactive marrow changes in the pedicle, or abnormal wedging of the posterior aspect of the vertebral body was retrospectively determined. This proportion was then compared with the proportion of patients in whom spondylolysis was correctly diagnosed by the initial interpreters of the MR images, who used only direct visualization of defects of the pars interarticularis to make the diagnosis. RESULTS: Twenty (30%) of 66 levels of lumbar spondylolysis were misdiagnosed when the MR images were initially interpreted using direct visualization of defects of the pars interarticularis. An increased sagittal diameter of the spinal canal was the most common ancillary observation, occurring at 60 of 66 levels of lumbar spondylolysis. This finding was present in all patients with grade II, III, or IV spondylolisthesis, in 95% of patients with grade I spondylolisthesis; and in 77% of patients with no anterolisthesis. Thirty-two (48%) of 66 lumbar levels showed wedging of the posterior aspect of the vertebral body, which correlated significantly with the grade of spondylolisthesis. Reactive marrow changes in the pedicle distinct from normal adjacent levels were seen on MR images in 24(36%) of 66 levels of lumbar spondylolysis. On MR images, 97% of all levels of lumbar spondylolysis yielded one or more ancillary observations, including all 20 of the cases originally misdiagnosed. CONCLUSION: The combined use of ancillary observations and direct visualization of pars interarticularis defects makes MR imaging effective in revealing lumbar spondylolysis. PMID- 9207532 TI - Brain abscess in hereditary hemorrhagic telangiectasia. PMID- 9207533 TI - A systematic approach for interpreting MR images of the seizure patient. AB - A systematic approach needs to be used to review MR scans in epilepsy patients to avoid the common pitfalls engendered by the subtle nature of many epileptogenic lesions. One should always evaluate the hippocampus regardless of other MR findings to avoid missing dual abnormalities. False-positive and false-negative diagnosis of hippocampal sclerosis can be avoided by evaluating the hippocampus after correcting for head rotation [by assessing the internal auditory canals and atria). Periventricular heterotopia can be successfully diagnosed by systematically studying the periventricular regions, especially those adjacent to the atria of the lateral ventricles. Gray matter lateral to the ventricles (excluding the caudate nucleus) is always an abnormal finding. Sulcal and cortical morphologic abnormalities are particularly difficult to diagnose unless a high index of suspicion is maintained. Cortical thickening is indicative of a developmental anomaly and should be screened in an organized manner. Because epilepsy is generally a cortical process, one must search for subtle cortical abnormalities, including focal atrophic abnormalities and lesions without mass effect. Diligence will offer its own rewards. PMID- 9207534 TI - Abdominal case of the day. Bouveret's syndrome. PMID- 9207535 TI - Abdominal case of the day. Focal liver enhancement on contrast-enhanced CT scan caused by obstruction of the superior vena cava (SVC). PMID- 9207536 TI - Abdominal case of the day. Recurrent pyogenic cholangitis (RPC). PMID- 9207537 TI - Abdominal case of the day. Malrotation of the small bowel. PMID- 9207539 TI - Angiography/interventional case of the day. Splenic artery pseudoaneurysm associated with pancreatitis. PMID- 9207538 TI - Angiography/interventional case of the day. Fractured central venous catheter. PMID- 9207540 TI - Angiography/interventional case of the day. Arteriogenic impotence from posttraumatic bilateral proximal cavernosal artery occlusions. PMID- 9207541 TI - Angiography/interventional case of the day. Polyarteritis nodosa (PAN). PMID- 9207542 TI - Chest case of the day. Morgagni's hernia. PMID- 9207543 TI - Chest case of the day. Scimitar syndrome. PMID- 9207544 TI - Chest case of the day. Extramedullary hematopoiesis (EMH). PMID- 9207545 TI - Chest case of the day. Histiocytosis X. PMID- 9207546 TI - Head and neck case of the day. Schwannoma of the right facial nerve. PMID- 9207547 TI - Head and neck case of the day. Thyroglossal duct cyst. PMID- 9207548 TI - Head and neck case of the day. Dermoids of the submandibular space. PMID- 9207549 TI - Head and neck case of the day. Carotid body tumor. PMID- 9207550 TI - Musculoskeletal case of the day. Pseudoaneurysm of the left superficial femoral artery. PMID- 9207551 TI - Musculoskeletal case of the day. Lymphoma of skeletal muscle. PMID- 9207552 TI - Musculoskeletal case of the day. Chronic recurrent multifocal osteomyelitis (CRMO). PMID- 9207553 TI - Neuroradiology case of the day. Dysembryoplastic neuroepithelial tumor. PMID- 9207555 TI - Neuroradiology case of the day. CNS cryptococcal infection. PMID- 9207557 TI - Neuroradiology case of the day. Multiple cerebral abscesses associated with isolated pulmonary arteriovenous malformation. PMID- 9207554 TI - Neuroradiology case of the day. Leptomeningeal melanocytosis. PMID- 9207558 TI - Pediatric case of the day. Streptococcal myositis and perianal cellulitis. PMID- 9207559 TI - Pediatric case of the day. Duchenne's pseudohypertrophic muscular dystrophy. PMID- 9207560 TI - Pediatric case of the day. Pulmonary sling with tracheal stenosis and bronchus suis. PMID- 9207561 TI - Pediatric case of the day. Parovarian cysts. PMID- 9207562 TI - Klippel-Trenaunay and Parkes-Weber syndromes. PMID- 9207563 TI - Informed consent: recall versus comprehension. PMID- 9207564 TI - Renal leiomyosarcoma mimicking transitional cell carcinoma. PMID- 9207565 TI - A new film for black-and-white slides. PMID- 9207566 TI - Re: Percutaneous transpulmonary CT-guided liver biopsy. PMID- 9207567 TI - MR imaging of a pericardial gossypiboma. PMID- 9207569 TI - When is a diagnostic mammogram a screening mammogram? PMID- 9207568 TI - "Mild" and "slight". PMID- 9207570 TI - Posttraumatic intrapericardial diaphragmatic hernia. PMID- 9207571 TI - Reopening of the ductal ampulla during balloon angioplasty of native aortic coarctation. PMID- 9207572 TI - Anthrax meningoencephalitis: radiologic findings. PMID- 9207573 TI - Precaval right renal artery: have you seen this? PMID- 9207574 TI - Cutaneous nodules, pain, and thrombophlebitis as an adverse reaction to gadolinium contrast media. PMID- 9207575 TI - Fertility after chemotherapy for testicular germ cell cancer. AB - OBJECTIVE: To investigate the impact of cytostatic chemotherapy on long-term fertility in patients with testicular germ cell cancer. BACKGROUND: Many patients with testicular germ cell cancer show impaired spermatogenesis before undergoing cytotoxic chemotherapy. The known infertility before treatment and the reversibility of the fertility problems observed in some of them after successful anticancer treatment so far have prevented an assessment of the true impact of chemotherapy on long-term fertility. The introduction of a wait-and-see strategy (surveillance) for patients with testicular cancer and recent prospective trials comparing patients with and without cytotoxic chemotherapy now have provided the means for estimating the extent to which chemotherapy itself affects long-term fertility. RESULT(S): Whether spermatogenesis is impaired irreversibly by chemotherapy is determined by the cumulative dose of cisplatin. At cumulative doses > 400 mg/m2, irreversible impairment of gonadal function should be expected. CONCLUSION(S): At cumulative cisplatin doses < 400 mg (equivalent to 4 courses of state-of-the-art treatment), chemotherapy is unlikely to cause irreversible damage to fertility. PMID- 9207576 TI - Randomized controlled trial of superovulation and insemination for infertility associated with minimal or mild endometriosis. AB - OBJECTIVE: To evaluate the efficacy of superovulation and IUI versus no treatment for infertility associated with minimal or mild endometriosis. DESIGN: Randomized trial. SETTING(S): London Health Sciences Centre, University Campus, The University of Western Ontario, London, Ontario; and Royal Victoria Hospital, McGill University, Montreal, Quebec, Canada. PATIENT(S): Three hundred eleven cycles in 103 couples in whom minimal or mild endometriosis was the sole identified subfertility factor. INTERVENTION(S): Superovulation with FSH and IUI. MAIN OUTCOME MEASURE(S): Live birth. RESULT(S): Live birth followed 14 of 127 (11%) superovulation and IUI cycles and 4 of 184 (2%) no-treatment cycles. The odds ratio was 5.6 (95% confidence interval 1.8 to 17.4) in favor of superovulation and IUI. CONCLUSION(S): Treatment with superovulation and IUI was associated with superior outcome both by crude live-birth rates and proportional hazard analysis. PMID- 9207577 TI - Endometriosis-associated pelvic pain: evidence for an association between the stage of disease and a history of chronic pelvic pain. AB - OBJECTIVE: To track the severity and location of pelvic pain associated with endometriosis throughout the reproductive-age years and to evaluate the association between these pain parameters and the stage of disease. DESIGN: Historical prospective study. SETTING: Tertiary care center. PATIENT(S): Forty eight women with endoscopically staged endometriosis and chronic pelvic pain who had undergone medical and/or conservative surgical therapy. INTERVENTION(S): Each participant was administered a questionnaire that included a determination of the severity and location of her pain. MAIN OUTCOME MEASURE(S): The stage of disease, the area of the pelvis that contained the bulk of disease, the severity of pain, and the location of the most severe pain were recorded. RESULT(S): The mean duration from the initial diagnosis until follow-up was 15.7 +/- 3.1 years, Twenty-one (43.8%) subjects denied any symptoms of pain on follow-up evaluation. Of the 27 patients with persistent pain, 21 (78%) identified the location of their most severe pain as being the same as at initial diagnosis. The stage of disease at initial diagnosis was significantly associated with a higher degree of pain at follow-up. CONCLUSION(S): These data suggest that endometriosis associated chronic pelvic pain commonly persists throughout the reproductive years and that endometriosis stage is directly related to the persistence of pelvic pain. PMID- 9207578 TI - Correlation between endometriosis-associated dysmenorrhea and the presence of typical or atypical lesions. AB - OBJECTIVE: To evaluate the correlation between the severity of endometriosis associated dysmenorrhea and the extent of the disease assessed both with a current classification system and with the number and type of endometriosis lesions. DESIGN: Prospective, blinded study. SETTING: Tertiary care, university hospital. PATIENT(S): Sixty-five consecutive patients with endometriosis diagnosed at laparoscopy performed for pelvic pain, infertility, or adnexal mass. INTERVENTION(S): The patients were submitted preoperatively to a questionnaire including the assessment of the severity of dysmenorrhea by means of a 10-point linear analog scale. Evaluation of all visible signs of endometriosis at laparoscopy was performed by a surgeon not aware of the patients' symptoms. MAIN OUTCOME MEASURE(S): The correlation between the severity of dysmenorrhea and the total score for endometriosis and the partial scores for superficial, deep, and adhesion disease as assessed with a current classification system was evaluated. The severity of dysmenorrhea was also correlated with the total number of superficial implants and with the number of typical, pigmented versus atypical, nonpigmented lesions. RESULT(S): The linear analog score for dysmenorrhea correlated significantly with the total endometriosis score, the partial score for deep endometriosis, and the partial score for adhesions. There was no correlation between the pain score for dysmenorrhea and the partial score for superficial endometriosis, nor with the total number of endometriosis implants, the number of typical implants, or the number of atypical implants. CONCLUSION(S): The current classification system for endometriosis, devised primarily to formulate a prognosis in infertile patients, correlates significantly with endometriosis-associated dysmenorrhea. The present study does not support the suggested association between atypical implants and the severity of dysmenorrhea. PMID- 9207579 TI - Distribution of heat shock proteins in eutopic and ectopic endometrium in endometriosis and adenomyosis. AB - OBJECTIVE: To evaluate the pathophysiologic role of heat shock proteins and to examine the effect of danazol on these proteins in patients with endometriosis and adenomyosis. DESIGN: Immunohistochemical identification of human heat shock proteins 27, 60, and 70 in endometrial glandular cells identified using monoclonal antibodies. SETTING: Department of obstetrics and gynecology in a university hospital. PATIENT(S): Subjects were 119 women with documented endometriosis or adenomyosis. The subjects were divided into three groups: fertile control (n = 38), with 14 in the proliferative phase and 24 in the secretory phase; endometriosis (n = 38); and adenomyosis (n = 43), including 33 who underwent hysterectomy and 10 treated with danazol. MAIN OUTCOME MEASURE(S): Staining of glandular cells by semiquantitative immunostaining (evaluation nomogram) score. RESULT(S): Significantly increased expression of heat shock protein 27 was noted in eutopic endometrium from patients with endometriosis and adenomyosis as compared with controls, regardless of the menstrual phase. The scores for heat shock protein 27 and heat shock protein 70 in the ectopic endometrium of patients with endometriosis were low compared with those in eutopic endometrium, whereas in adenomyosis, the scores were similar to those of eutopic endometrium. After treatment with danazol, the expression of heat shock proteins returned to control levels. CONCLUSION(S): We suggest that abnormally increased expression of heat shock proteins plays a role in the pathophysiology of endometriosis and adenomyosis. PMID- 9207580 TI - Goserelin acetate (Zoladex) plus endometrial ablation for dysfunctional uterine bleeding: a large randomized, double-blind study. AB - OBJECTIVE: To confirm the advantages of goserelin prior to endometrial ablation for the treatment of dysfunctional uterine bleeding. DESIGN: Multicenter, prospective, randomized, double-blind study. PATIENT(S): Cycling premenopausal women with dysfunctional uterine bleeding. TREATMENT: Patients were randomized to goserelin or placebo (sham depot) once monthly for 2 months prior to endometrial ablation. Treatment was timed to allow surgery 6 weeks later on day 7 of the menstrual cycle. MAIN OUTCOME MEASURE(S): Amenorrhea rates, endometrial histology and thickness, pain and blood loss scores, and surgical parameters. RESULT(S): At 24 weeks after surgery, significantly more goserelin than placebo patients experienced amenorrhea (40% versus 26%). Blood loss was reduced from baseline, but not different between the groups. At surgery, mean endometrial thickness was 1.6 mm and 3.4 mm for the goserelin and placebo groups, respectively, with significantly more atrophic glands and stroma in the goserelin group. Surgery was significantly shorter (by 22%) and easier in the goserelin than in the placebo group, with a significantly lower median fluid absorption in the goserelin groups. In both groups, pain scores were reduced patient satisfaction was high (> 92%), and re-intervention rate was low (2.8%). CONCLUSION(S): Goserelin in combination with endometrial ablation was superior to endometrial ablation alone for the treatment of dysfunctional uterine bleeding. PMID- 9207581 TI - Reduction of adhesion formation by postoperative administration of ionically cross-linked hyaluronic acid. AB - OBJECTIVE: To examine the efficacy of various formulations of hyaluronic acid (HA), including HA ionically cross-linked with trivalent iron, in animal models of adhesion formation. DESIGN: Hyaluronic acid formulation of varying concentrations and cross-linked densities were prepared and evaluated in a rabbit uterine horn model and a rabbit sidewall model. SETTING: ETHICON, Inc., Somerville, New Jersey. SUBJECT(S): New Zealand White rabbits. INTERVENTION(S): Test formulations were applied as intraperitoneal instillates after surgery. MAIN OUTCOME MEASURE(S): Adhesion formation was assessed at 7 and 14 days (sidewall and uterine horn model, respectively). RESULT(S): Hyaluronic acid that was not ionically cross-linked was ineffective in reducing adhesions in these models even at high viscosity, whereas the ionically cross-linked formulations of HA with trivalent iron were highly effective. Efficacy improved with increased levels of ionic cross-linking. Flowable gels, which could be delivered readily by syringe and cannula, also were effective when administered at a site remote from injury and with saline present. CONCLUSION(S): Whereas previous studies show that HA was effective in reducing adhesions peripheral to the site of injury, HA ionically cross-linked with trivalent iron was effective in reducing adhesions at all sites. From these studies, a formulation of HA ionically cross-linked with trivalent iron, 0.5% Ferric Hyaluronate Gel (LUBRICOAT; ETHICON, Inc., Somerville, NJ), was identified for subsequent clinical evaluations. PMID- 9207582 TI - Clomiphene citrate ovulation induction in combination with a timed intrauterine insemination: the value of urinary luteinizing hormone versus human chorionic gonadotropin timing. AB - OBJECTIVE: To evaluate the clinical pregnancy rates (PRs) in anovulatory, male factor, and unexplained infertility using clomiphene citrate (CC) with an IUI and to evaluate the difference in PRs between urinary LH testing and hCG administration for timing of the IUI. DESIGN: Retrospective clinical study. SETTING: Academic, tertiary care fertility center. PATIENT(S): One hundred thirty eight couples (432 cycles) undergoing IUI with CC ovulation induction as a treatment for unexplained, anovulatory, or male factor infertility were selected. INTERVENTION(S): All women with unexplained or male factor infertility received CC at a dose of 50 mg/d, and those with anovulation received CC at a dose ranging from 50 to 200 mg/d. All women in the study received a single IUI either the morning after a urinary LH surge or 36 to 38 hours after an evening hCG injection. MAIN OUTCOME MEASURE(S): Clinical PR. RESULT(S): There were no differences in the clinical PRs between LH testing or hCG administration in any of the three groups. Clinical PRs were extremely low in the male factor infertility group regardless of the timing used. CONCLUSION(S): These data suggest that the success of IUI with CC is not dependent on the method used to establish the timing for the IUI. In couples undergoing IUI with CC, the use of urinary LH testing may result in lower costs by reducing patient visits and the midcycle ultrasound. PMID- 9207583 TI - Increased follicular fluid total and free cortisol levels during the luteinizing hormone surge. AB - OBJECTIVE: To determine the changes in follicular fluid (FF) total and free cortisol during the LH surge in naturally ovulating women. PATIENT(S): Twenty-six women having diagnostic laparoscopy during the follicular phase of normal menstrual cycles were selected. INTERVENTION(S): Blood samples were collected 1 day before, the day of, and 1 day after surgery and the results of serum E2 and LH were used to divide the cycles retrospectively into pre- and post-LH surge groups. Follicular fluid was collected during laparoscopy. MAIN OUTCOME MEASURE(S): Serum P, total and free cortisol, and FF volume, E2, P, total cortisol, and free cortisol were measured on the day of surgery. RESULT(S): Median serum and FF P levels were significantly higher in the post-LH surge group compared with the pre-LH surge group (0.54 versus 1.54 ng/mL [1.7 versus 4.85 nmol/L] and 5.03 versus 28.0 micrograms/mL [15.8 versus 88.0 mumol/L], respectively). Follicular fluid volume also increased significantly after the surge (2.5 versus 4.5 mL). Median serum total and free and percent free cortisol were higher after the surge, although not significantly. In contrast, FF total, free, and percent free levels increased dramatically between pre- and post-LH surge samples (4.41 versus 43.6 ng/mL [16.0 versus 158 nmol/L], 0.138 versus 6.68 ng/mL [0.5 versus 24.2 nmol/L], and 3.3% versus 15.0%, respectively; P < 0.05). CONCLUSION(S): An increase in total and free cortisol occurs in the follicle during the LH surge. Cortisol and its regulation by 11 beta-hydroxysteroid dehydrogenase therefore may exert a physiologic role in oocyte maturation or ovulation. PMID- 9207584 TI - Chromosome analysis of aborted conceptuses of recurrent aborters positive for anticardiolipin antibody. AB - OBJECTIVE: To elucidate the relationship between anticardiolipin antibody and recurrent abortion. DESIGN: Prospective clinical study. SETTING: Institutional practice in which patients with recurrent abortion were registered at the outpatient clinic for infertility of Niigata University Hospital. PATIENT(S): Five hundred sixty-one patients with recurrent abortions and 148 patients who were not recurrent aborters and who had experienced sporadic abortion. INTERVENTION(S): Aborted conceptuses for chromosome analyses were collected from the patients. MAIN OUTCOME MEASURE(S): The positive rate of anticardiolipin antibody was assessed in patients with recurrent abortion. Chromosome analyses of aborted conceptuses were performed in 10 patients with positive anticardiolipin antibody who had experienced another pregnancy that resulted in repeated abortion. Similar analyses of aborted conceptuses from 148 sporadic early abortions (controls) were performed. RESULT(S): The incidence of chromosome abnormalities in anticardiolipin antibody-positive recurrent aborters was 20.0% (2 of 10 cases), which was significantly lower than that of patients with sporadic abortion (60.1%, 89 of 148 cases). CONCLUSION(S): The low incidence of chromosome abnormalities in aborted conceptuses of patients with positive anticardiolipin antibody suggests that this antibody is strongly implicated in the genesis of recurrent abortions. PMID- 9207585 TI - Intracervical and fundal administration of levonorgestrel for contraception: endometrial thickness, patterns of bleeding, and persisting ovarian follicles. AB - OBJECTIVE(S): To study the prevalence of persisting ovarian follicles and to assess the endometrial changes and patterns of vaginal bleeding over 1 year of use of a 20 micrograms/24 h levonorgestrel-releasing intracervical contraceptive device. DESIGN: Prospective, randomized study. SETTING: Two family planning clinics in Helsinki, Finland. PATIENT(S): Women requesting intrauterine hormonal contraception. INTERVENTION(S): Insertion of a levonorgestrel-releasing intracervical contraceptive device into the cervical canal (group 1, n = 151) or fundally into the uterine cavity (group 2, n = 147) for contraception. MAIN OUTCOME MEASURE(S): Transvaginal ultrasonography of the ovaries and endometrium at insertion and 3, 6, and 12 months after insertion. Data on bleeding were collected using menstrual diary cards. RESULTS: Persisting ovarian follicles were found in < 8% of women. In both groups, the amount of endometrial tissue decreased significantly in 3 months. The incidence of amenorrhea during the 1st year was higher in the fundal insertion group. CONCLUSION(S): The number of persisting follicles was low. Follicles resolved within 6 to 8 weeks. No association was found between persisting follicles and problems of bleeding. Compared with intracervical insertion, fundal insertion resulted in more uniform endometrial suppression and fewer days of bleeding and spotting. PMID- 9207586 TI - Hydrosalpinx fluid enhances human trophoblast viability and function in vitro: implications for embryonic implantation in assisted reproduction. AB - OBJECTIVE: To assess the effects of hydrosalpinx fluid on human cytotrophoblast viability and function in vitro. DESIGN: Human cytotrophoblasts obtained from third-trimester placentas were cultured in vitro with hydrosalpinx fluid, and cell viability and protein production were assayed. SETTING: A university hospital. PATIENT(S): Ten hydrosalpinx fluid samples obtained from seven women with clearly diagnosed hydrosalpinges. INTERVENTION(S): Recovery of hydrosalpinx fluid by transvaginal aspiration or at the time of surgery. MAIN OUTCOME MEASURE(S): Cell viability was assessed by the XTT assay. Secretion of trophoblast oncofetal fibronectin (tropho-uteronectin) and beta-hCG by cultured trophoblasts was determined by Western blot and ELISA of the culture media. RESULT(S): With increasing concentrations of hydrosalpinx fluid from 0% to 20%, there was a significant increase in trophoblast cell viability (1.63-fold increase in 20% hydrosalpinx fluid). Likewise, both Western blot and ELISA assays demonstrated a significant increase in tropho-uteronectin production by trophoblasts with increasing hydrosalpinx fluid concentrations (3.76-fold increase in 20% hydrosalpinx fluid). beta-Human chorionic gonadotropin production also increased significantly in the presence of hydrosalpinx fluid (3.31-fold increase in 20% hydrosalpinx fluid). CONCLUSION(S): These findings suggest that hydrosalpinx fluid improves human trophoblast viability in vitro and enhances the production of tropho-uteronectin and beta-hCG by these cells. PMID- 9207587 TI - Velocity of endometrial wavelike activity in spontaneous cycles. AB - OBJECTIVE: To analyze the velocity and wave intervals of endometrial wavelike activity in spontaneous menstrual cycles. DESIGN: Prospective observational ultrasound (US) evaluation of endometrial wavelike activity. SETTING: University hospital. MAIN OUTCOME MEASURE(S): Endometrial wavelike activity, wave velocity, wave intervals. PATIENT(S): Twenty-three menstrual cycles were evaluated by frequent US investigations in 16 healthy, regularly cycling female volunteers. INTERVENTION(S): Transvaginal US examination of endometrial wavelike activity. RESULT(S): Nineteen cycles proved to be ovulatory. Wave velocity and wave intervals were calculated in waves from fundus to cervix and in waves from cervix to fundus. The velocity of waves from fundus to cervix increased from the midfollicular phase to the late follicular phase. Waves from cervix to fundus showed their highest velocity in the periovulatory period. CONCLUSION(S): Velocity of endometrial wavelike activity reached a peak in the periovulatory phase, whereas the wave intervals were shortest in that phase. PMID- 9207588 TI - Immunodetection of cotinine protein in granulosa-lutein cells of women exposed to cigarette smoke. AB - OBJECTIVE: To detect immunoreactivity to cotinine protein, a major metabolite of nicotine, in granulosa-lutein cells from patients exposed to cigarette smoke, as measured by levels of cotinine in follicular fluid (FF) samples. DESIGN: Controlled immunocytochemical study. SETTING: Hospital IVF-ET program treating infertile patients. PATIENT(S): Twenty-eight women classified by self-reported smoking habits: active smokers (n = 17), passive smokers (n = 4), and nonsmokers (n = 7). INTERVENTION(S): Ovarian hyperstimulation. MAIN OUTCOME MEASURE(S): Grades of immunostaining intensity were assessed in granulosa-lutein cells. Patient scores of cell immunostaining were calculated and regressed on levels of FF cotinine. RESULT(S): Cotinine levels in FF were higher in active smokers than in passive smokers or nonsmokers. Cotinine immunostaining was visualized in the nucleus and cytoplasm of granulosa-lutein cells. Mean grades and mean scores of immunostaining intensity were higher in active smokers than in passive smokers or nonsmokers. There was a strong positive correlation between scores of cell immunostaining and FF cotinine levels. CONCLUSION(S): The association between cotinine expression in granulosa-lutein cells and FF cotinine provides reliable evidence for a dose-related effect. This constituent of cigarette smoke appears to interact directly with and incorporate into these ovarian cells. Our approach seems useful for monitoring ovarian exposure to environmental toxins. PMID- 9207589 TI - Stigma, disclosure, and family functioning among parents of children conceived through donor insemination. AB - OBJECTIVE: To examine the influence of gender, male infertility factor, and other demographic variables on stigma and whether parents tell their children that they were conceived by donor insemination (DI) and to ascertain if stigma and the disclosure decision affect parental bonding with the child or the quality of the interparental relationship. DESIGN: One hundred eighty-four San Francisco Bay Area couples who had become parents by DI were asked to complete a self administered questionnaire. SETTING: A private infertility practice. PATIENT(S): Eighty-two men and 94 women who completed the questionnaire. MAIN OUTCOME MEASURE: A questionnaire assessing disclosure, stigma, parental bonding, and the quality of the interparental relationship. RESULT(S): Factors that increased the couple's likelihood of disclosure included younger age, azoospermia, lower stigma scores, and having more than one DI child. Fathers who scored higher on stigma reported less parental warmth and parental fostering of independence. CONCLUSION(S): Because the decision regarding disclosure of DI treatment was not linked to parental bonding with the child or to the quality of the interparental relationship, we cannot conclude that nondisclosure is harmful to family relationships or is a symptom of family problems. The husband's perceptions of stigma however, may affect the father--child relationship adversely. PMID- 9207590 TI - The feasibility of assessing women's perceptions of the risks and benefits of fertility drug therapy in relation to ovarian cancer risk. AB - OBJECTIVE: To determine the feasibility of asking women undergoing fertility treatment the maximum increased risk of ovarian cancer they would be willing to tolerate in order to take ovulation-induction drugs. DESIGN: A prospective pilot study of women attending fertility clinics over a 2-month period. SETTING: Two tertiary care fertility clinics in Toronto. PATIENT(S): Sixty-one English speaking women were approached and 85% (n = 52) were enrolled. INTERVENTION(S): A self-administered questionnaire with fertility-specific questions. Thirty-eight women also were asked to complete standardized scales of anxiety and optimism. MAIN OUTCOME MEASURE(S): Women's report of the maximum level of lifetime risk of ovarian cancer they were willing to tolerate in order to undergo fertility treatment. RESULT(S): Seventy-nine percent were willing to accept an increased risk of ovarian cancer. Only 24% understood that treatment for ovarian cancer usually was not curative. CONCLUSION(S): A majority of patients were willing to tolerate a modest increase in their lifetime risk of ovarian cancer because of fertility treatment, most basing their estimate of acceptable risk on limited awareness of the issue. PMID- 9207591 TI - Age at natural menopause in a population-based screening cohort: the role of menarche, fecundity, and lifestyle factors. AB - OBJECTIVE: To verify whether a population-based hypothesis (age at menarche and age at natural menopause have an inverse relationship) also applies at the level of the individual and to investigate what other factors predict age at natural menopause. DESIGN: Prospective cohort study (the Doorlopend Onderzoek Morbiditeit/Mortaliteit [DOM] project). SETTING: Prevention Breast Cancer Screening Centre, Utrecht, The Netherlands. PATIENT(S): A cohort of 3,756 Dutch women, born between 1911 and 1925, participating in a population-based breast cancer screening program, who experienced a natural menopause. Three samples of women were studied: a sample who did not use oral contraceptives (OCs) (n = 3,347), a sample of OC users (n = 409), and a combined sample of OC users and nonusers (n = 3,756). MAIN OUTCOME MEASURE(S): Age at menopause and menarche, fertility patterns, OC use, height, weight, smoking, and demographic variables. RESULT(S): No relation was found between age at menarche and age at natural menopause. The total percentage of variance in age at natural menopause explained by multiple regression including all factors was minimal, ranging from 1.3% to 9.7% in OC users. Linear regression analysis indicated a slight secular trend in age at menopause. CONCLUSION(S): Frisch's hypothesis could not be corroborated at the individual level. These results suggest that age at menarche and menopause should be treated as independent risk factors for breast cancer. Modification of age at menopause by lifestyle factors (except possibly for OC use) appears minimal. PMID- 9207592 TI - Effect of hormone replacement therapy on growth hormone stimulation in women with premature ovarian failure. AB - OBJECTIVE: To assess the effect of oral hormone replacement therapy (HRT) on body weight, insulin-like growth factor I (IGF-I), and GH response to exogenous GHRH [corrected] in women with premature ovarian failure (POF) [corrected]. DESIGN: Controlled clinical study. SETTING: Outpatients studied in the department of endocrinology of the University Hospital in Vienna. PATIENT(S): Twenty-four women with POF (study group) and 24 volunteers with normal ovarian cycles (control group). INTERVENTION(S): Pituitary GHRH [corrected] stimulation was performed in all women at study entry and in patients with POF after 1, 6, and 12 months of standard oral HRT. Blood samples were collected from 15 minutes before to 120 minutes after GHRH administration [corrected]. Body weight also was evaluated. RESULT(S): No differences in baseline and stimulated serum GH were found either between POF women and controls or in POF women during HRT. Women with POF without HRT had significantly higher IGF-I levels; a reduction in circulating IGF-I levels occurred during HRT. Body weight remained stable. CONCLUSION(S): Our results show the following: [1] Women with POF have similar Gh secretion patterns as healthy age-matched women; [2] physiologic HRT has no impact on GHRH-induced [corrected] GH stimulation; and [3] HRT has no impact on body weight in women with POF. PMID- 9207593 TI - Fertilization and pregnancy rates after intracytoplasmic sperm injection using ejaculate semen and surgically retrieved sperm. AB - OBJECTIVE: To compare the fertilization rates and pregnancy rates (PRs) in intracytoplasmic sperm injection (ICSI) using sperm from ejaculates of normal and abnormal semen, epididymal sperm, and testicular sperm of obstructive and nonobstructive azoospermic patients. DESIGN: Retrospective study. SETTING: The Egyptian IVF-ET Center. PATIENT(S): Three hundred fifty patients underwent 366 ICSI cycles. INTERVENTION(S): ICSI, epididymal sperm aspiration, and testicular biopsy. MAIN OUTCOME MEASURE(S): Fertilization rates and PRs. RESULT(S): Patients were divided into five groups according to the quality and source of sperm. Patients in group 1 underwent 102 cycles of ICSI using ejaculated abnormal semen, group 2 underwent 44 cycles using epididymal sperm, group 3 underwent 82 cycles using testicular sperm from obstructive azoospermia, group 4 underwent 80 cycles using testicular sperm from nonobstructive azoospermia, and group 5 underwent 58 cycles using normal semen. There was no significant difference in the fertilization rates and PRs among groups 1, 2, and 3. In group 4, the fertilization rate and PR were significantly lower than in all other groups. In group 5, the fertilization rate was significantly higher than in all other groups. CONCLUSION(S): The fertilizing ability of sperm in ICSI is highest with normal semen and lowest with sperm extracted from a testicular biopsy in nonobstructive azoospermia. There was no significant difference in fertilization rates and PRs between ejaculated sperm of different parameters and surgically retrieved sperm in obstructive azoospermia. PMID- 9207594 TI - Intracytoplasmic sperm injection for treating infertility associated with sperm autoimmunity. AB - OBJECTIVE: To determine whether intracytoplasmic sperm injection (ICSI) can be used to achieve normal fertilization, embryo cleavage, and pregnancies in cases of sperm autoimmunity. DESIGN: A retrospective analysis of ICSI results in sperm antibody-positive and randomly selected antibody-negative groups. SETTING: University- and hospital-based reproductive research laboratory and tertiary referral IVF program. PATIENT(S): Thirty-nine couples selected on the basis of a strongly positive result for sperm antibodies of immunoglobulin (Ig) G and/or IgA immunoglobulin class in the male partner and a control group of 140 antibody negative couples. INTERVENTION(S): Human menopausal gonadotropin, hCG and Lucrin (Abbott Australasia, Kurnell, NSW, Australia) were given by injection. Oocyte collection was by transvaginal ovarian puncture. Blood was collected for beta-hCG measurement. MAIN OUTCOME MEASURE(S): Normal fertilization, embryo cleavage, establishment of clinical pregnancy, and delivery. RESULT(S): There were no significant differences in fertilization rates (62% versus 58%) or clinical pregnancy rates (19% versus 12%) between sperm antibody-positive and sperm antibody-negative patient groups. CONCLUSION: Intracytoplasmic sperm injection is an effective treatment for patients with severe sperm autoimmunity. PMID- 9207595 TI - Failure of oocyte activation after intracytoplasmic sperm injection using round headed sperm. AB - OBJECTIVE: To examine the outcome of intracytoplasmic sperm injection (ICSI) with round-headed sperm (globozoospermia). DESIGN: Retrospective analysis. SETTING: In vitro fertilization laboratory with extensive ICSI experience. PATIENT(S): A patient couple with infertility because of globozoospermia seeking ICSI treatment. MAIN OUTCOME MEASURE(S): Fertilization, cleavage, and pregnancy rates. INTERVENTION(S): Intracytoplasmic sperm injection and calcium ionophore. RESULT(S): This couple experienced only 7% fertilization after ICSI in their first cycle. Treatment of the unfertilized oocytes with calcium ionophore 20 hours after ICSI-induced fertilization and cleavage of 70% of the oocytes. Embryo quality was fair to good. On the second cycle, 8 of the injected oocytes were treated with ionophore immediately after ICSI and the remaining 20 oocytes were untreated. Normal fertilization was achieved in 75% of the treated and 10% of the untreated oocytes. Treatment of these unfertilized oocytes with ionophore 20 hours after ICSI resulted in fertilization in 73%. Pregnancy was not achieved after either ICSI cycle. Ultrastructural analysis indicated multiple structural abnormalities in the sperm. CONCLUSION(S): These results indicate that the round headed sperm from this patient were incapable of oocyte activation after ICSI. This may be the reason for the frequent ICSI fertilization failure seen with this condition. Current ICSI procedures may not always overcome the infertility associated with globozoospermia, and further study of the etiology of this condition is needed. PMID- 9207596 TI - Effect of age and cycle responsiveness in patients undergoing intracytoplasmic sperm injection. AB - OBJECTIVE: To evaluate the effects of age and number of embryos available for transfer on pregnancy rate (PR) in couples undergoing intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective study of patients undergoing ICSI for male factor infertility. SETTING: Tertiary care academic center. PATIENT(S): One hundred twelve patients < 37 years of age and 57 patients > or = 37 years of age who underwent ICSI with uterine ET or tubal ET. INTERVENTION: Compare cycles in which three embryos or fewer were available for transfer with those with four or more available embryos. MAIN OUTCOME MEASURE(S): Implantation, embryo availability, and pregnancy and miscarriage rates. RESULT(S): Younger patients did significantly better with regard to PR (47% versus 26%), implantation rate (11.4% versus 6.6%), and ongoing PR (40% versus 19%). Patients in whom more than four embryos were transferred also did better than patients in whom three or fewer embryos were available for transfer, with an implantation rate of 14.2% versus 4.2%. In women < 37 years of age, 85% of cycles produced more than three embryos, versus 61% in women > or = 37 years of age. When cycles yielding three embryos or fewer were excluded, the younger group tended to do better, with an ongoing PR of 48% versus 33% for the older group, but the differences were not statistically significant. Both groups had similar number of embryos transferred per cycle. CONCLUSION(S): Age affects cycle responsiveness, and the number of embryos available for transfer affects fertility in patients undergoing ICSI. Older women with good ovarian response, producing more than three embryos suitable for transfer, have a PR similar to younger patients. Cycles yielding three embryos or fewer have a poor prognosis. PMID- 9207597 TI - Office laparoscopy under local anesthesia for gamete intrafallopian transfer: technique and tolerance. AB - OBJECTIVE: To describe our technique for laparoscopic GIFT under local anesthesia and to evaluate patient tolerance and surgeon satisfaction in 175 consecutive procedures. DESIGN: Prospective cohort study. SETTING: University infertility practice. PATIENT(S): All GIFT candidates from 1992 to 1996 were offered the procedure. Of 119 patients, 119 chose local anesthesia for 175 procedures and 1 patient elected to have general anesthesia. INTERVENTION(S): Transvaginal ultrasound-guided egg retrieval followed by GIFT in the clinic procedure room with a 5-mm laparoscope and two accessory 3-mm trocars with local anesthesia and i.v. sedation. MAIN OUTCOME MEASURE(S): Patient tolerance and acceptance, duration of the procedure, amount of analgesics, surgeon satisfaction, and pregnancy rate (PR). RESULT(S): The laparoscopic portion lasted an average of 27 minutes, with a mean dose of 1.41 mg of midazolam and 68 micrograms of fentanyl used. Sixty-nine percent of the patients scored "very good," 20% "good," 9% "acceptable," and 2% "poor." All 38 patients undergoing 97 repeat procedures selected local anesthesia again. For women < 40 years of age, clinical PR and delivery rate were 43% and 38%, respectively. CONCLUSION(S): Routine office GIFT under local anesthesia is effective and well accepted by the surgeon and is preferred by patients. It offers a significant cost containment and scheduling flexibility in addition to high success rates. PMID- 9207598 TI - Elevated levels of interleukin-6 in the follicular fluid at the time of oocyte retrieval for in vitro fertilization may predict the development of early-form ovarian hyperstimulation syndrome. AB - OBJECTIVE: To determine the possible predictive role of interleukin-2 (IL-2), IL 6, and tumor necrosis factor (TNF-alpha) in the development of early-form ovarian hyperstimulation syndrome (OHSS). DESIGN: Nested, case-control study. SETTING: An IVF unit, university-based program. PATIENT(S): Follicular fluid (FF) was obtained from 322 high responders. The study group and control group comprised 10 patients who developed early, severe OHSS and 10 who did not develop OHSS, respectively. An additional control group included 10 low-responder patients who did not develop OHSS. INTERVENTION(S): Ovulation induction with hMG combined with GnRH analogue. MAIN OUTCOME MEASURE(S): All FF samples were tested for IL-2, IL 6, and TNF-alpha. The patient's serum was tested for mean E2 and P concentrations. RESULT(S): Interleukin-6 levels in the FF were significantly higher in the OHSS group than in the two control groups, whereas no differences were found in IL-2 and TNF-alpha. No correlation was found between the FF concentrations of IL-2, IL-6, and TNF-alpha and the mean serum E2 levels or the number of oocytes retrieved. CONCLUSION(S): Elevated levels of IL-6 in the preovulatory FF at the time of oocyte retrieval for IVF may predict the development of early-form OHSS in high responders. PMID- 9207599 TI - Recombinant follicle-stimulating hormone (follitropin beta, Puregon) yields higher pregnancy rates in in vitro fertilization than urinary gonadotropins. AB - OBJECTIVE: To assess ongoing pregnancy rates (PRs) in IVF after treatment with recombinant FSH (follitropin beta, Puregon; NV Organon, Oss, The Netherlands) as compared with urinary gonadotropins. DESIGN: A combined analysis of three prospective, multicenter, randomized, comparative trials. SETTING: Twenty-five IVF centers in 13 countries. PATIENT(S): Six hundred ninety-seven infertile women receiving recombinant FSH and 463 women receiving hMG or urinary FSH and undergoing one cycle of controlled ovarian hyperstimulation and IVF-ET. INTERVENTION(S): A center-based and study-based analysis weighing the treatment differences in individual centers and studies, respectively. MAIN OUTCOME MEASURES(S): Pregnancy rate at least 12 weeks after ET per started cycle. RESULTS(S): In the center-based analysis, the ongoing PR was 22.9% for recombinant FSH and 17.9% for urinary gonadotropins. The 5.0% treatment difference (95% confidence interval [CI], 0.2% to 9.7%) was significant. When the results of the cryoprogram were included, the treatment difference increased to 6.4% (95% CI, 1.4% to 11.3%). Also in the study-based analysis, significantly higher PRs were seen after follitropin beta treatment. CONCLUSION(S): Follitropin beta (Puregon) used for controlled ovarian hyperstimulation in IVF yields significantly higher PRs compared with urinary gonadotropins. PMID- 9207600 TI - An intrauterine insemination-ready cryopreservation method compared with sperm recovery after conventional freezing and post-thaw processing. AB - OBJECTIVE: To test a sucrose-glycerol cryoprotectant for IUI-ready sperm preparation. DESIGN: Semen aliquots from normozoospermic donors either were subjected to conventional semen freezing (TES and Tris yolk buffer in 7.4% final glycerol) with post-thaw processing or were preprocessed and frozen in HEPES buffered human tubal fluid with 1% human serum albumin, 4% sucrose, and 6% glycerol. All aliquots were cooled to 4 degrees C, exposed to liquid nitrogen vapors, and stored in liquid nitrogen. Aliquots from each were processed by centrifugation resuspension or by centrifugation in Percoll (Pharmacia, Alameda, CA) before sperm parameters were analyzed. SETTING: University-based andrology laboratory. MAIN OUTCOME MEASURE(S): Recovery of motile sperm. RESULT(S): Percoll processing produced preparations with higher percentages of motile cells; however, cryopreserved sperm had a lower recovery of motile sperm compared with Percoll-processed fresh semen or centrifugation/resuspension-processed fresh or frozen samples. The percentages of sperm with normal morphologies were significantly increased in the IUI-ready samples compared with samples frozen conventionally. The IUI-ready Percoll-processed sample produced the best results, with a final mean motility of 36% and an overall yield of motile sperm of 17.4%. CONCLUSION(S): The sucrose-glycerol-based cryoprotectant produced an IUI-ready preparation with motile sperm recovery comparable to that of conventional semen cryopreservation but with improved percent morphology. PMID- 9207601 TI - Oligozoospermia induced by exogenous testosterone is associated with normal functioning residual spermatozoa. AB - OBJECTIVE: To determine the functional capacity of residual spermatozoa in semen samples from normal men with T enanthate-induced oligozoospermia. DESIGN: Prospective clinical study. SETTING: Academic research center. PATIENT(S): Twelve healthy men were studied while participating in a multicenter T enanthate contraceptive efficacy study. Data were analyzed from only eight subjects, whose sperm concentrations were between 1.3 and 10 x 10(6)/mL at the suppression phase. INTERVENTION(S): Testosterone enanthate (200 mg) was administered IM weekly during the suppression and treatment (efficacy) phases (total 15 months). MAIN OUTCOME MEASURE(S): Sperm function tests (stimulated acrosome reaction, sperm hyperactivation [HA], and zona-free hamster oocyte penetration tests) were performed during the pretreatment, suppression (usually after 6 to 10 weeks of treatment, when sperm concentration was anticipated to decrease to < 10 x 10(6)/mL), and recovery phases. Studies were not done during the contraceptive efficacy phase because only one of the subjects was not azoospermic. RESULT(S): Mean sperm concentration was reduced but sperm motility, motility characteristics, and morphology were not affected by T enanthate treatment. The residual spermatozoa in the ejaculate could acrosome react, exhibited normal HA, and maintained the capacity to penetrate and fuse with the oocyte. CONCLUSION(S): Suppression of spermatogenesis to moderate oligozoospermia (< 10 x 10(6)/mL) with exogenous T enanthate administration was not associated with impaired sperm function of the residual spermatozoa. The study did not exclude the possibility that disorders of sperm function might occur when spermatogenesis is suppressed further to very severe oligozoospermia (< 1 x 10(6)/mL), commonly observed in hormonal male contraceptive clinical trials. PMID- 9207602 TI - Nonenzymatic antioxidant activity of human seminal plasma. AB - OBJECTIVE: To examine the nonenzymatic antioxidant properties of human seminal plasma. DESIGN: Determination of total (peroxyl) radical-trapping antioxidant parameter and malondialdehyde concentration in seminal plasma of 112 men attending IVF-ET or intracytoplasmic sperm injection (ICSI) programs. SETTING: University-based center for reproductive medicine. PATIENT(S): Normozoospermic men, ages 25 to 50 years, of fertility proved by a pregnancy resulting from IVF ET, and men ages 25 to 50 years with negative IVF-ET or ICSI outcome, distinguished by their smoking status. INTERVENTION(S): Seminal plasma was obtained from ejaculates collected for IVF-ET or ICSI procedures. MAIN OUTCOME MEASURE(S): Total (peroxyl) radical-trapping antioxidant parameter and malondialdehyde concentrations. RESULT(S): No significant differences for total (peroxyl)-radical-trapping antioxidant parameter were observed between normozoospermic IVF-ET or asthenozoospermic ICSI patients; neither was a difference revealed for pregnancy-positive or -negative couples within those groups. Age, smoking status, or increased leukocyte count had no effect on total (peroxyl) radical-trapping antioxidant parameter or malondialdehyde concentrations in seminal plasma. CONCLUSION(S): Nonenzymatic antioxidant activity in seminal plasma does not reflect sperm fecundity ability. PMID- 9207603 TI - Detection using antisperm monoclonal antibodies of shared epitopes expressed by human spermatozoa and oocytes. AB - OBJECTIVE: To determine whether human spermatozoa and oocytes share common antigenic epitopes, supporting the hypothesis that their cross-linking by antisperm antibodies present in the clinical sera of infertile couples could promote sperm adhesion to the oolemma. DESIGN: Human and hamster eggs were studied for the presence of antigens recognized by a panel of World Health Organization Task Force monoclonal antibodies (mAbs) originally raised against human spermatozoa. A new technique was devised, using frozen sections of paraformaldehyde-fixed individual human and hamster eggs, to screen rapidly antisperm mAbs for egg reactivity. Living zona-free human and hamster eggs then were exposed to Covaspheres (Duke Scientific, Palo Alto, CA) coupled with these mAbs to document the presence of reactive epitopes on the oolemma. SETTING: Academic research environment. MAIN OUTCOME MEASURE(S): Indirect immunofluorescence and Covasphere rosetting. RESULT(S): Eleven of 37 antisperm mAbs tested reacted with fixed hamster eggs and 10 reacted with human eggs. Five of 6 mAbs reactive with both fixed eggs also reacted with the oolemma of living, zona-free eggs. CONCLUSION(S): Common antigenic epitopes, some of which are shared with somatic tissues, exist on the oolemma of human eggs and on the plasma membrane of human spermatozoa. PMID- 9207604 TI - The effect of the oviduct, uterine, and in vitro environments on zona thinning in the mouse embryo. AB - OBJECTIVE: To evaluate the impact of the oviduct, uterine, and in vitro environments on zona pellucida thinning in the mouse embryo. DESIGN: Female mice were stimulated with pregnant mare serum gonadotropin and mated and hCG injection. Unilateral oviduct ligation was performed on day 2 of gestation using the dorsal approach. The mice were divided into equal groups and killed on days 2, 3, 4, 5, and 10 of gestation. In vitro incubated embryos served as controls. Average daily zona thickness measurements were subjected to analysis of variance and paired Student's t-test. SETTING: The laboratory of the assisted reproductive program of Rush University Medical Center. MAIN OUTCOME MEASURE(S): Progressive daily decrease in average zona thickness. RESULT(S): Zona measurements of embryos flushed out of uterine horns, ligated oviducts, and in vitro incubation demonstrated statistically significant decreases in zona thickness, from 9.6 +/- 1.6 microns (day 3) to 6.0 +/- 0.8 microns (day 5), from 11.6 +/- 2.2 microns (day 2) to 6.0 +/- 1.6 microns (day 5), and from 11.1 +/- 2.0 microns (day 2) to 6.0 +/- 1.6 microns (day 5), respectively. There were no differences in average zona thickness for embryos in the same cell stage and same protocol day in all three locations. CONCLUSION(S): Zona thinning seems to be induced primarily by the dividing embryo before implantation. A substantial tubal and uterine contribution to zona thinning was not detected in this mouse embryo model. PMID- 9207605 TI - Identification of early pregnancy landmarks by transvaginal sonography: analysis by logistic regression. AB - OBJECTIVE: To assess the feasibility of logistic regression analysis for determining the gestational ages at which detection of early pregnancy landmarks first can be observed. DESIGN: Retrospective analysis. SETTING: University-based tertiary care clinic. PATIENT(S): Eighty-two women with viable singleton pregnancies in whom ovulation had been achieved by an injection of hCG. INTERVENTION(S): Two hundred fifteen transvaginal sonographic scans. MAIN OUTCOME MEASURE(S): Logistic regression was used to estimate the probability of detection of sonographic findings as a function of gestational age. RESULT(S): We found that the likelihood of visualization of a gestational sac or fetal heart motion could be represented accurately by logistic equations. Gestational age at which there was 95% probability of visualization was 35.5 days for the gestational sac and 44.5 days for fetal cardiac activity. The probability of detecting fetal cardiac activity was 95% when the mean gestational sac diameter was 1.6 cm and was 99% at 1.9 cm. CONCLUSION(S): The sonographic appearances of developmental landmarks in early pregnancy occurs within well-defined gestational time periods, and the probabilities for visualization can be closely approximated using a logistic model. Our results suggest that the number of sonographic examinations required to document infertility treatment success can be minimized by surveillance at standardized gestational ages. PMID- 9207606 TI - The role of in vitro fertilization and intracytoplasmic sperm injection in couples with unexplained infertility after failed intrauterine insemination. AB - OBJECTIVE: To determine an optimal insemination technique in patients undergoing IVF after failed IUI and the role of intracytoplasmic sperm injection (ICSI) in such cases. DESIGN: Prospective, randomized study in couples with unexplained infertility (n = 63) and mild endometriosis (n = 7) undergoing IVF after four IUI cycles. Sibling oocytes were randomized into standard IVF or ICSI insemination according to the order of retrieval. SETTING: In vitro fertilization program at the Instituto Valenciano de Infertilidad, Valencia, Italy. PATIENT(S): Seventy couples with unexplained infertility undergoing IVF after failing to conceive with controlled ovarian stimulation and IUI. INTERVENTION(S): In vitro fertilization and ICSI. MAIN OUTCOME MEASURE(S): Fertilization, cleavage, and embryo quality were compared in IVF- and ICSI-inseminated oocytes. RESULT(S): There was no significant difference in fertilization rates between ICSI (60.4%) and conventional IVF (54.0%). Similarly, there was no difference in embryo quality between both groups. There was no total fertilization failure in ICSI inseminated oocytes, whereas 8 (11.4%) of 70 cases showed absence of fertilization when conventional IVF was used. CONCLUSION(S): Couples with unexplained infertility and mild endometriosis failing to conceive with IUI and undergoing IVF have an 11.4% chance of fertilization failure that can be overcome easily by using ICSI in at least some oocytes. ICSI, however, is not superior to IVF as an insemination technique in most cases. These data should be used in counseling patients. PMID- 9207607 TI - Bilateral internal jugular venous thrombosis complicating severe ovarian hyperstimulation syndrome after prophylactic albumin administration. AB - OBJECTIVE: To report a case of bilateral thrombosis of the internal jugular veins in a patient after controlled ovarian hyperstimulation and IVF. DESIGN: Case report. SETTING: University-based IVF program. INTERVENTION(S): Ovulation induction with gonadotropins, IVF-ET, albumin administration, and therapeutic heparinization. MAIN OUTCOME MEASURE(S): Doppler ultrasound of neck veins. RESULT(S): Severe ovarian hyperstimulation syndrome and bilateral thrombosis of the internal jugular veins occurred despite prophylactic administration of albumin. This was treated successfully with therapeutic heparinization. CONCLUSION(S): Internal jugular venous thrombosis, a rare complication of ovulation induction with gonadotropins, should be considered in patients with neck pain and swelling. PMID- 9207608 TI - The best measure of uncertainty--hypothesis tests or tests of estimation? PMID- 9207609 TI - Definition of ureteral endometriosis? PMID- 9207610 TI - Need: a special Medline--for prior to 1966. PMID- 9207611 TI - Approach to "complete endometrial sclerosis". PMID- 9207612 TI - Detection of CAH heterozygotes. PMID- 9207613 TI - Detection of CAH heterozygotes. PMID- 9207614 TI - "The clomid twins": waiting for single isomer heaven. PMID- 9207615 TI - Evaluation of diastolic filling of left ventricle in health and disease: Doppler echocardiography is the clinician's Rosetta Stone. AB - Abnormalities of diastolic function have a major role in producing the signs and symptoms of heart failure. However, diastolic function of the heart is a complex sequence of multiple interrelated events, and it has been difficult to understand, diagnose and treat the various abnormalities of diastolic filling that occur in patients with heart disease. Recently, Doppler echocardiography has been used to examine the different diastolic filling patterns of the left ventricle in health and disease, but confusion about diagnosis and treatment options has arisen because of the misinterpretation of these flow velocity curves. This review presents a simplified approach to understanding the process of diastolic filling of the left ventricle and interpreting the Doppler flow velocity curves as they relate to this process. It has been hypothesized that transmitral flow velocity curves show a progression over time with diseases involving the myocardium. This concept can be applied clinically to estimate left ventricular filling pressures and to predict prognosis in selected groups of patients. Specific therapy for diastolic dysfunction based on Doppler flow velocity curves is discussed. PMID- 9207616 TI - Estimating mean pulmonary wedge pressure in patients with chronic atrial fibrillation from transthoracic Doppler indexes of mitral and pulmonary venous flow velocity. AB - OBJECTIVES: We sought to obtain a noninvasive estimation of mean pulmonary wedge pressure (MPWP) in patients with chronic atrial fibrillation (AF). BACKGROUND: It has previously been demonstrated that MPWP can be reliably estimated from Doppler indexes of mitral and pulmonary venous flow (PVF) in patients with sinus rhythm. Doppler estimation of MPWP has not been validated in patients with AF. METHODS: MPWP was correlated with variables of mitral and pulmonary venous flow velocity as assessed by Doppler transthoracic echocardiography in 35 consecutive patients. The derived algorithm was prospectively tested in 23 additional patients. RESULTS: In all patients the mitral flow pattern showed only a diastolic forward component. A significant but relatively weak correlation (r = -0.50) was observed between MPWP and mitral deceleration time. In 12 (34%) of 35 patients, the pulmonary vein flow tracing demonstrated only a diastolic forward component; a diastolic and late systolic forward flow was noted in the remaining 23 patients (66%). A strong negative correlation was observed between MPWP and the normalized duration of the diastolic flow (r = -0.80) and its initial deceleration slope time (r = -0.91). Deceleration time > 220 ms predicted MPWP < or = 12 mm Hg with 100% sensitivity and 100% specificity. When estimating MPWP by using the equation MPWP = -94.261 PVF deceleration time -9.831 Interval QRS to onset of diastolic PVF -16.337 Duration of PVF + 44.261, the measured and predicted MPWP closely agreed with a mean difference of -0.85 mm Hg. The 95% confidence limits were 4.8 and -6.1 mm Hg. CONCLUSIONS: In patients with chronic AF, MPWP can be estimated from transthoracic Doppler study of PVF velocity signals. PMID- 9207617 TI - Effect of beta-blockade on mortality in patients with heart failure: a meta analysis of randomized clinical trials. AB - OBJECTIVES: We sought to evaluate the current evidence for an effect of beta blockade treatment on mortality in patients with congestive heart failure (CHF). BACKGROUND: Although numerous small studies have suggested a benefit with beta blocker therapy in patients with heart failure, a clear survival benefit has not been demonstrated. A recent combined analysis of several studies with the alpha- and beta-adrenergic blocking agent carvedilol demonstrated a significant survival advantage; however, the total number of events was small. Furthermore, it is unclear if previous studies with other beta-blockers are consistent with this finding. METHODS: Randomized clinical trials of beta-blockade treatment in patients with CHF from January 1975 through February 1997 were identified using a MEDLINE search and a review of reports from scientific meetings. Studies were included if mortality was reported during 3 or more months of follow-up. RESULTS: We identified 35 reports, 17 of which met the inclusion criteria. These studies included 3,039 patients with follow-up ranging from 3 months to 2 years. Beta blockade was associated with a trend toward mortality reduction in 13 studies. When all 17 reports were combined, beta-blockade significantly reduced all-cause mortality (random effect odds ratio [OR] 0.69, 95% confidence interval [CI] 0.54 to 0.88). A trend toward greater treatment effect was noted for nonsudden cardiac death (OR 0.58, 95% CI 0.40 to 0.83) compared with sudden cardiac death (OR 0.84, 95% CI 0.59 to 1.2). Similar reductions in mortality were observed for patients with ischemic (OR 0.69, 95% CI 0.49 to 0.98) and nonischemic cardiomyopathy (OR 0.69, 95% CI 0.47 to 0.99). The survival benefit was greater for trials of the drug carvedilol (OR 0.54, 95% CI 0.36 to 0.81) than for noncarvedilol drugs (OR 0.82, 95% CI 0.60 to 1.12); however, the difference did not reach statistical significance (p = 0.10). CONCLUSIONS: Pooled evidence suggests that beta-blockade reduces all-cause mortality in patients with CHF. Additional trials are required to determine whether carvedilol differs in its effect from other agents. PMID- 9207618 TI - Role of cytokines in the mechanism of action of amlodipine: the PRAISE Heart Failure Trial. Prospective Randomized Amlodipine Survival Evaluation. AB - OBJECTIVES: We sought to determine whether the beneficial effects of amlodipine in heart failure may be mediated by a reduction in tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) levels. We postulated that TNF-alpha and IL 6 levels may also have predictive value in patients with congestive heart failure (CHF). BACKGROUND: The molecular mechanism for progression of CHF may involve cytokine overexpression. The effect of amlodipine on cytokine levels in patients with CHF is unknown. METHODS: In the Prospective Randomized Amlodipine Survival Evaluation (PRAISE) trial, we used enzyme-linked immunosorbent assay to measure plasma levels of TNF-alpha in 92 patients and IL-6 in 62 patients in New York Heart Association functional classes III and IV randomized to receive amlodipine (10 mg/day) or placebo. Blood samples were obtained for cytokine measurement at baseline and at 8 and 26 weeks after enrollment. RESULTS: The baseline amlodipine and placebo groups did not differ in demographics and cytokine levels. Mean (+/- SD) plasma levels of TNF-alpha were 5.69 +/- 0.32 pg/ml, and those of IL-6 were 9.23 +/- 1.26 pg/ml at baseline. These levels were elevated 6 and 10 times, respectively, compared with those of normal subjects (p < 0.001). Levels of TNF alpha did not change significantly over the 26-week period (p = 0.69). However, IL-6 levels were significantly lower at 26 weeks in patients treated with amlodipine versus placebo (p = 0.007 by the Wilcoxon signed-rank test). An adverse event-CHF or death-occurred more commonly in patients with higher IL-6 levels. CONCLUSIONS: Amlodipine lowers plasma IL-6 levels in patients with CHF. The beneficial effect of amlodipine in CHF may be due to a reduction of cytokines such as IL-6. PMID- 9207619 TI - Patients with mild heart failure worsen during withdrawal from digoxin therapy. AB - OBJECTIVES: We investigated whether patients with mild heart failure due to left ventricular systolic dysfunction were at risk of worsening during digoxin withdrawal. BACKGROUND: Deterioration during digoxin withdrawal is often believed to be restricted to patients with moderate to severe clinical evidence of heart failure. To test this hypothesis, we studied the outcome of patients categorized by treatment assignment and a clinical signs and symptoms heart failure score in two rigorously designed clinical heart failure trials: the Prospective Randomized Study of Ventricular Function and Efficacy of Digoxin (PROVED) and the Randomized Assessment of Digoxin and Inhibitors of Angiotensin-Converting Enzyme (RADIANCE) trial. METHODS: Potential differences in treatment failure, left ventricular ejection fraction and exercise capacity were evaluated in three groups of patients: those with mild heart failure (score < or = 2) who were withdrawn from digoxin (Dig WD Mild); those with moderate heart failure (score > 2) who were withdrawn from digoxin (Dig WD Moderate); and patients who continued receiving digoxin regardless of heart failure score (Dig Cont). RESULTS: Heart failure score at randomization did not predict outcome during follow-up in Dig Cont-group patients. Dig WD Mild-group patients were at increased risk of treatment failure and had deterioration of exercise capacity and left ventricular ejection fraction compared with that in Dig Cont-group patients (all p < 0.01). Patients in the Dig WD Moderate group were significantly more likely to experience treatment failure than patients in either the Dig WD Mild or Dig Cont group (both p < 0.05). CONCLUSIONS: Patients with systolic left ventricular dysfunction were at risk of clinical deterioration after digoxin withdrawal despite mild clinical evidence of congestive heart failure. PMID- 9207620 TI - Effect of inhaled nitric oxide on normal human left ventricular function. AB - OBJECTIVES: This study determined the effects of inhaled nitric oxide (NO) on load-independent indexes of normal human left ventricular (LV) function. BACKGROUND: Inhaled NO is a potent and selective pulmonary vasodilator. However, when it is used in patients with congestive heart failure, the decrease in pulmonary vascular resistance (PVR) is often associated with an increase in pulmonary capillary wedge pressure. NO has been shown to have a negative inotropic action, but it is not known whether it affects LV chamber function when delivered by inhalation. METHODS: Eleven subjects (51 to 69 years old) with normal LV function (mean ejection fraction 72% [range 60% to 80%]) were studied. Four patients had concomitant coronary artery disease. Pressure-volume loop recordings were used to determine end-systolic and end-diastolic pressure-volume and preload recruitable stroke work relations. NO was delivered at 20 ppm for 10 min. In an additional group of patients with normal LV function, PVR (n = 5) and NO metabolites (n = 9) were measured. RESULTS: There was no effect of inhaled NO on steady state LV pressures, volumes, contractility, contraction duration, active relaxation (time constant of relaxation, peak negative first derivative of left ventricular pressure), diastolic compliance or PVR. NO metabolites (methemoglobin and nitrate) were present in the LV cavity at the same concentration as right atrial venous blood, suggesting inactivation of free NO before arrival in the LV chamber. This study had a power of 0.995 to detect a 5% change in contractility (slope of preload recruitable stroke work relation) for alpha = 0.05, based on the multiple linear regression model used. CONCLUSIONS: These results indicate that 20 ppm of inhaled NO does not have significant effects on normal LV function. This lack of effect may be due in part to rapid inactivation of free NO in transit to the heart. PMID- 9207621 TI - Comparison of electron beam computed tomography with intracoronary ultrasound and coronary angiography for detection of coronary atherosclerosis. AB - OBJECTIVES: This analysis compared the results of electron beam computed tomography (EBCT) with those of coronary angiography and intracoronary ultrasound (ICUS) for the in vivo detection of coronary atherosclerotic plaques. BACKGROUND: EBCT is a new imaging modality for identification of coronary calcifications. Coronary angiography depicts advanced changes in coronary morphology, whereas ICUS is an established diagnostic tool that detects the early stages of coronary artery disease. METHODS: In 57 patients (54 +/- 9 years old), 267 coronary segments were analyzed with EBCT (3-mm slices, acquisition time 100 ms, threshold definition of coronary calcification at 130 Hounsfield units in an area > or = 1 mm2, Agatston calcium score), coronary angiography and ICUS. The analysis was based on the number and extent of coronary calcifications on EBCT, coronary lumen reduction on coronary angiography and plaque formation with and without ultrasound signs of calcifications on ICUS. RESULTS: Compared with coronary angiography, EBCT yielded a sensitivity of 66%, a specificity of 78%, a positive predictive value of 39% and a negative predictive value of 91%. Compared with ICUS, EBCT yielded an overall sensitivity of 66%, a specificity of 88% and an overall accuracy of 81%. For plaques with and without ultrasound signs of calcifications, the sensitivity of EBCT was 97% and 47%, specificity 80% and 75% and overall accuracy 82% and 69%, respectively. CONCLUSIONS: This in vivo correlation between ICUS and EBCT demonstrates that EBCT is a noninvasive method that helps to visualize the atherosclerotic process by localization and quantification of coronary artery calcifications. EBCT detects calcified plaques with high accuracy. Plaques without ultrasound signs of calcifications can be detected by EBCT but with lower sensitivity but equivalent specificity. PMID- 9207622 TI - Flow-function relation in patients with chronic coronary artery disease and reduced regional function. A positron emission tomographic and two-dimensional echocardiographic study with coronary vasodilator stress. AB - OBJECTIVES: We sought to elucidate the flow-function relation in chronic postischemic dysfunction during vasodilator stress. BACKGROUND: In patients with ischemia and regional dysfunction, stress echocardiography can elicit three responses in the dysfunctioning segments: no change, improvement or worsening. The physiology underlying these responses is unclear. METHODS: Seventeen patients with ischemia and left ventricular dysfunction underwent evaluation of regional function by two-dimensional echocardiography and myocardial blood flow by positron emission tomography and 13N-ammonia. Flow (ml/min per g) and function (regional wall motion score [RWMS] from 1 = normal to 4 = dyskinetic) were evaluated both at rest and after dipyridamole (0.56 mg/kg body weight over 4 min). RESULTS: In 45 normal segments, rest to dipyridamole flow increased from 0.83 +/- 0.22 (mean +/- 1 SD) to 1.87 +/- 0.90 (p < 0.01) with a hyperkinetic contraction pattern. Among dysfunctioning segments, responders (n = 11) showed an upsloping flow-function curve during stress (i.e., increased function [RWMS rest 2.5 +/- 0.5 vs. dipyridamole 1.2 +/- 0.4] and increased flow [rest 0.69 +/- 0.30 vs. dipyridamole 1.89 +/- 1.43, p < 0.01]); nonresponders (n = 20) had a flat flow-function curve during dipyridamole (i.e., fixed function [RWMS rest and dipyridamole 2.6 +/- 0.5] and no flow increase [rest 0.64 +/- 0.24 vs. dipyridamole 0.87 +/- 0.51, p = NS): Ischemic segments (n = 9) exhibited a downsloping flow-function curve during dipyridamole (i.e., worsened function [RWMS rest 2 +/- 0.5, dipyridamole 3.1 +/- 0.6] and no significant flow change [rest 0.67 +/- 0.29 vs. dipyridamole 0.79 +/- 0.23, p = NS]). CONCLUSIONS: Myocardial segments with rest dysfunction and a contractile reserve elicitable by a vasodilator stress more often exhibit residual flow reserve, whereas segments with a fixed or worsening mechanical response during stress show a flat flow response. PMID- 9207623 TI - Prognostic significance of spontaneous echo contrast in the thoracic aorta: relation with accelerated clinical progression of coronary artery disease. AB - OBJECTIVES: The purposes of this study were to identify the incidence of aortic smoke in an unselected cohort of patients and to determine the utility of this measurement as a clinical marker for future coronary events and long-term cardiac prognosis. BACKGROUND: Although spontaneous echo contrast detected within the cardiac chambers has been associated with an increased risk of thromboembolism, less is known about "smoke" within the thoracic aorta and its relation to progression of coronary artery disease. METHODS: We prospectively assessed 118 unselected, consecutive male patients (mean age 67 years, range 29 to 86) who underwent transesophageal echocardiography (TEE). The presence of aortic smoke was identified by swirling echodense shadows distinct from high gain artifact. A positive result required confirmation by two of three independent observers. RESULTS: Aortic smoke without dissection was found in 25 of the patients (21%). Indications for TEE, coronary risk factors, the incidence of reduced left ventricular ejection fraction and mitral insufficiency and known coronary artery disease severity collectively did not differ significantly at baseline between the groups with and without smoke. Follow-up averaged 20.4 months (range 18 to 24) and was 100% complete for mortality and 98% complete for morbidity. The presence of aortic smoke was an independent predictor of myocardial infarction (16.0% vs. 2.2%, p < 0.005) and cardiac death (20.0% vs. 1.1%, p < 0.0001). These statistics remained significant after covarying for age, ejection fraction < 50%, hypertension, diabetes, aortic dimension, the presence of an atheromatous plaque and smoke in the left atrium. CONCLUSIONS: Spontaneous echo contrast detected within the thoracic aorta by transesophageal echocardiography is a common and important clinical marker that is strongly associated with an increased risk for future myocardial infarction and cardiac mortality. Future studies will attempt to define the pathophysiology of this relation and assess whether aggressive revascularization strategies and antithrombotic therapy may aid in the reduction of this risk. PMID- 9207624 TI - Remodeling after myocardial infarction in humans is not associated with interstitial fibrosis of noninfarcted myocardium. AB - OBJECTIVES: This study was specifically designed to evaluate whether noninfarcted hypertrophic myocardium in patients with end-stage heart failure after myocardial infarction (MI) is associated with an increase in interstitial fibrous tissue. BACKGROUND: Postinfarction remodeling consists of complex alterations that involve both infarcted and noninfarcted myocardium. The question arises whether ventricular dysfunction is due to physical events, such as inadequate myocardial hypertrophy to compensate for increased tangential wall stress, or is caused by the development of progressive interstitial fibrosis in noninfarcted myocardium. METHODS: Fifteen hearts were obtained as cardiac explants (n = 13) or at autopsy (n = 2) from patients with end-stage coronary artery disease. Sixteen normal hearts served as reference hearts. Samples were taken from the left ventricular (LV) wall that contained the infarcted area, the border area and noninfarcted myocardium remote from scar areas. Collagen was quantified biochemically and microdensitophotometrically. Collagen type I and III ratios were analyzed by using the cyanogen bromide method and immunohistochemical staining, followed by microdensitophotometric quantification. RESULTS: In noninfarcted myocardium remote from the scar areas, total collagen levels and collagen type I/III ratios did not differ statistically from those in reference hearts. These observations contrasted with high total collagen content and high collagen type I/III ratios in scar and border areas. CONCLUSIONS: Remodeling of LV myocardium after MI in patients with end-stage heart failure is not necessarily associated with interstitial fibrosis in noninfarcted hypertrophic myocardium remote from scar areas. This finding raises questions regarding therapeutic interventions designed to prevent or retard the development of interstitial fibrosis. PMID- 9207625 TI - Use of exercise echocardiography for prognostic evaluation of patients with known or suspected coronary artery disease. AB - OBJECTIVES: This study prospectively compared the incremental prognostic benefit of exercise echocardiography with that of exercise testing in a large cohort. BACKGROUND: Exercise echocardiography is widely accepted as a diagnostic tool, but the prognostic information provided by this test, incremental to clinical and stress testing evaluation, is ill-defined. METHODS: Clinical, exercise and echocardiographic variables were studied in a consecutive group of 500 patients undergoing exercise echocardiography. After exclusion of patients who underwent revascularization within 3 months of the stress test (n = 16, 3%) and those lost to follow-up (n = 21, 4%), the remaining 463 patients (mean [+/-SD] age 57 +/- 12 years, 302 men) were followed-up for 44 +/- 11 months. Outcome was related to the exercise and echocardiographic findings, and the incremental prognostic benefit of exercise echocardiography was compared with that of standard exercise testing. RESULTS: Cardiac events occurred in 81 patients (17%), including 33 (7%) with spontaneous events (cardiac death, myocardial infarction and unstable angina) and 48 with late revascularizations due to progressive symptoms. In a multivariate Cox proportional hazards model, the likelihood of any cardiac event was increased in the presence of ischemia (relative risk [RR] 5.06, 95% confidence interval [CI] 3.09 to 8.29, p < 0.001) and lessened by more maximal stress, measured as percent age-predicted maximal heart rate (RR per 5% increment 0.84, 95% CI 0.77 to 0.92, p < 0.001). Spontaneous events were more strongly predicted by ischemia (RR 8.20, 95% CI 3.41 to 19.71, p < 0.001) and percent age-predicted maximal heart rate (RR per 5% increment 0.78, 95% CI 0.67 to 0.91, p < 0.001). An interactive logistic regression model showed that the addition of echocardiographic to exercise and clinical data offered incremental predictive value. CONCLUSIONS: The presence of ischemia on the exercise echocardiogram can predict whether patients will experience an event. This relation is independent of, and incremental to, clinical and exercise data. PMID- 9207626 TI - Platelet-dependent thrombin generation in patients with hyperlipidemia. AB - OBJECTIVES: We evaluated coagulability as determined by platelet-dependent thrombin generation in hypercholesterolemic patients before and after treatment with pravastatin and in hypertriglyceridemic patients to investigate the usefulness of coagulability as an index of atherosclerosis and to determine the importance of treating hyperlipidemia. BACKGROUND: An understanding of the interaction between platelets and the plasma coagulation system is important for clarifying the mechanism of the procoagulant process. METHODS: We assessed coagulability in 58 patients with hypercholesterolemia (serum total cholesterol level > or = 220 mg/dl, age 56.5 +/- 1.5 years [mean +/- SEM]), 37 patients with hypertriglyceridemia (serum triglyceride level > or = 200 mg/dl, age 59.5 +/- 1.7 years), 13 patients with hypercholesterolemia plus hypertriglyceridemia (age 51.4 +/- 3.1 years) and 75 normal subjects (age 52.2 +/- 1.7 years). We also studied platelet-dependent thrombin generation in patients with hypercholesterolemia before and after treatment with pravastatin. Calcium chloride was added to 0.5 ml of platelet-rich plasma (150 x 10(9)/liter) to initiate coagulation. Ten microliters of the sample was transferred into 90 microliters of 3.8% sodium citrate at 10-min intervals for 30 min. A chromogenic substrate, S-2238, was added to each sample, and absorbance was measured spectrophotometrically at a wavelength of 405 nm to determine thrombin generation. RESULTS: Platelet dependent thrombin generation was increased in patients with hypercholesterolemia and patients with hypercholesterolemia plus hypertriglyceridemia (p < 0.01) compared with patients with hypertriglyceridemia and control subjects. Treatment with pravastatin normalized thrombin generation. CONCLUSIONS: Hypercholesterolemia, but not hypertriglyceridemia, was associated with increased platelet-dependent thrombin generation. Pravastatin normalized the generation of thrombin. PMID- 9207627 TI - In vivo low density lipoprotein oxidation relates to coronary reactivity in young men. AB - OBJECTIVES: This study was undertaken to examine the relation of in vivo low density lipoprotein (LDL) oxidation and other lipid risk factors to coronary reactivity in normal subjects. BACKGROUND: Experimental studies have shown that oxidized LDL (ox-LDL) particles are injurious to the vascular wall by impairing its normal vasodilator function. METHODS: We used noninvasive positron emission tomographic (PET) imaging with intravenous dipyridamole to measure coronary flow reserve, a marker of coronary endothelial and smooth muscle function, in 30 healthy men (mean [+/-SD] age 34.4 +/- 3.2 years). As a marker of in vivo LDL oxidation, the autoantibody titer against ox-LDL was measured by the enzyme linked immunosorbent assay method. RESULTS: Plasma levels of autoantibody titer against ox-LDL were inversely associated with coronary flow reserve (r = -0.42, p = 0.023). High LDL cholesterol levels (above median > 3.0 mmol/liter) were associated with a low coronary flow reserve only in subjects expressing simultaneously high levels of ox-LDL titer (above median). Subjects with simultaneously high levels of LDL cholesterol and ox-LDL titer had lower coronary flow reserve values than subjects in other groups (3.89 vs. > 5.0 in other groups, p = 0.066). CONCLUSIONS: These data provide evidence for the role of ox LDL in affecting the coronary reactivity in vivo and support the concept that oxidative modification of LDL particles provides a mechanism by which high LDL concentrations exhibit injurious effects on the coronary vascular bed. PMID- 9207628 TI - Isolated defect of adenosine-mediated coronary vasodilation: functional evidence for a new microangiopathic entity. AB - OBJECTIVES: The present study describes an isolated defect of the coronary vasodilation in response to adenosine in five patients examined for clinically suspected coronary microangiopathy. BACKGROUND: Coronary microangiopathies can be defined functionally as dysregulation of the microcirculatory vasomotion. METHODS: The five patients were compared with 24 control subjects. Coronary flow velocity was measured with an intracoronary Doppler guide wire (0.018 in. [0.046 cm], 12 MHz) at rest and during intracoronary administration of adenosine (80 micrograms/min and 160 micrograms/min over 3 min each), papaverine (10-mg bolus) and acetylcholine (30 micrograms/min over 5 min). Diameters of the epicardial coronary arteries were measured by quantitative coronary angiography. RESULTS: All subjects (patients and control) exhibited angiographically normal epicardial coronary arteries and normal and comparable endothelium-independent and dependent vasomotion, as assessed with papaverine (mean [+/-SD]-relative coronary flow reserve 2.62 +/- 0.66 vs. 2.97 +/- 0.88, p = 0.32) and acetylcholine (volumetric coronary flow reserve 2.61 +/- 0.27 vs. 2.91 +/- 0.67, p = 0.58), respectively. Affected patients were identified by an isolated complete defect of the adenosine-mediated vasodilation compared with control subjects (relative coronary flow reserve in response to 80 micrograms/min of adenosine 1.08 +/- 0.17 vs. 2.45 +/- 0.74 [p < 0.001] and 160 micrograms/min of adenosine 1.03 +/- 0.15 vs. 2.89 +/- 0.65 [p < 0.001]). CONCLUSIONS: These findings are consistent with functional evidence for a new entity of a coronary microangiopathy affecting a subtype of the endothelium-independent vasomotion. PMID- 9207629 TI - Bradykinin-induced vasodilation of human coronary arteries in vivo: role of nitric oxide and angiotensin-converting enzyme. AB - OBJECTIVES: The present study aimed to determine the role of nitric oxide (NO) and angiotensin-converting enzyme (ACE) in bradykinin (BK)-induced dilation of human coronary arteries in vivo. BACKGROUND: BK, produced by way of the kinin kallikrein system, causes endothelium-dependent vasodilation. However, little is known about the mechanism of BK-induced dilation of coronary arteries in humans in vivo. METHODS: The effects of an inhibitor of NO synthesis and of an ACE inhibitor on BK-induced coronary vasodilation were examined in 20 patients who had no significant atherosclerotic stenosis in the artery under study. Lumen diameters of the large epicardial coronary arteries and coronary blood flow (CBF) were measured by quantitative coronary arteriography and intracoronary Doppler technique. RESULTS: Intracoronary infusion of BK (0.6 and 2.0 micrograms/min) increased coronary artery diameter and CBF with no change in arterial pressure or heart rate. The BK-induced increases in coronary artery diameter and CBF were significantly reduced (p < 0.01) after pretreatment with NG-monomethyl-L-arginine (200 mumol) and were significantly increased (p < 0.01) after pretreatment with enalaprilat (50 micrograms). CONCLUSIONS: BK-induced dilation of human large epicardial and resistance coronary arteries is mediated by NO and increased by prior ACE inhibition. PMID- 9207631 TI - Attenuation of severity of myocardial ischemia during repeated daily ischemic episodes. AB - OBJECTIVES: The purpose of this study was to assess whether the severity of myocardial ischemia would be attenuated by repeated daily ischemic episodes, recorded by ambulatory electrocardiographic monitoring (AEM). BACKGROUND: Repetitive ischemic episodes induced by brief coronary occlusions in animal experiments and in humans during balloon coronary angioplasty produce preconditioning. We wanted to assess whether this phenomenon also exists during daily ischemic episodes. METHODS: Twenty-one patients with known coronary artery disease and ischemia on exercise testing and AEM were requested to walk a distance known to have previously caused myocardial ischemia on three consecutive occasions. Walking time was approximately 15 min and was followed by 5 min of rest. RESULTS: Mean maximal heart rate during the three walks was similar; however, the mean maximal ST segment depression decreased significantly from 2.21 mm during the first walk to 1.61 mm and 1.43 mm, respectively, on the second and third walks (p = 0.001). Ischemia duration was also significantly reduced on the second and third walks by 56% from 514 to 228 and 254 s, respectively (p = 0.012). The heart rate at onset of ischemia (ischemic threshold) increased from 99 beats/min on the first walk to 101 beats/min on the second walk and to 106 beats/min on the third walk (p = 0.058). CONCLUSIONS: This study demonstrated attenuation of myocardial ischemia with an associated increase in ischemic threshold in patients with repeated and adjacent ischemic episodes. This form of myocardial protection is likely to be encountered in patients during ordinary activity and may represent the clinical counterpart of myocardial preconditioning. PMID- 9207630 TI - Chinese adults are less susceptible than whites to age-related endothelial dysfunction. AB - OBJECTIVES: We sought to assess the effects of aging on the endothelial physiology of a group of Chinese adults. BACKGROUND: Several studies have documented an association between aging and progressive arterial endothelial dysfunction in white subjects. We hypothesized that age-related endothelial dysfunction, an important event in atherosclerosis, might be less marked in southern Chinese subjects, in whom the prevalence of coronary heart disease is only approximately 20% of that in industrialized countries. METHODS: We studied endothelial function in 76 healthy adults aged 16 to 70 years: 38 Chinese from a village of 3,000 people in southern China and 38 white subjects from Sydney, Australia. In each ethnic group, there were 19 younger persons (16 to 40 years) and 19 older adults (55 to 70 years). None had evidence of diabetes, hypertension or clinical vascular disease or had ever been regular cigarette smokers. With the use of high resolution external vascular ultrasound, brachial artery diameter was measured at rest, after flow increase (causing endothelium-dependent dilation) and after sublingual nitroglycerin (an endothelium-independent dilator). RESULTS: Endothelium-dependent dilation was similar in young Chinese (mean +/- SD 8.3 +/- 2.5%), young whites (7.9 +/- 2.0%) and older Chinese (6.8 +/- 2.9%), but it was significantly impaired in older whites (1.8 +/- 2.5%, p < 0.001 by analysis of variance). On multivariate analysis, older age was associated with impaired endothelium-dependent dilation (p < 0.001) (independent of the effects of serum cholesterol, gender and vessel size) in the white but not in the Chinese subjects (p = 0.83). Nitroglycerin-induced dilation was not significantly different with aging in either ethnic group. CONCLUSIONS: Endothelium-dependent dilation is similar in the arteries of healthy young Chinese and white adults. With older age, however, Chinese subjects are less susceptible to impaired endothelial function. PMID- 9207632 TI - Nicotine patch therapy in smoking cessation reduces the extent of exercise induced myocardial ischemia. AB - OBJECTIVES: We sought to determine the effects of nicotine patch therapy, when used to promote smoking cessation, on myocardial ischemia in patients with coronary artery disease. BACKGROUND: Nicotine patches substantially increase quit rates among cigarette smokers, but their safety in patients with myocardial ischemia who are attempting to quit smoking is unknown. METHODS: This is a prospective study using exercise thallium-201 single-photon emission computed tomography (SPECT) to assess serial changes in the total and ischemic myocardial perfusion defect size at baseline while patients were smoking and during treatment with 14- and 21-mg nicotine patches. Entry criteria required that patients 1) smoked > or = 1 pack of cigarettes per day; 2) had known coronary artery disease; and 3) had myocardial ischemia (i.e., > or = 5% reversible perfusion defect) on SPECT. All patients performed symptom-limited treadmill exercise, and the baseline SPECT study served as its own control. We interpreted and computer quantified the SPECT images with no knowledge of the testing sequence. RESULTS: Thirty-six of the 40 enrolled patients had exercise SPECT at baseline and during treatment with at least 14-mg nicotine patches. These patients had an initial perfusion defect size of 17.5 +/- 10.6% while smoking an average of 31 +/- 11 cigarettes per day for 40 +/- 12 years. A significant reduction in the total perfusion defect size (p < 0.001) was observed from baseline (17.5 +/- 10.6%) to treatment with 14-mg (12.6 +/- 10.1%) and 21-mg (11.8 +/- 9.9%) nicotine patches. This reduction occurred despite an increase in treadmill exercise duration (p < 0.05) and higher serum nicotine levels (p < 0.001). There was a significant correlation between the reduction in defect size and exhaled carbon monoxide levels (p < 0.001) because patients reduced their smoking by approximately 74% during the trial. CONCLUSIONS: Nicotine patches, when used to promote smoking cessation, significantly reduce the extent of exercise-induced myocardial ischemia as assessed by exercise thallium-201 SPECT. PMID- 9207633 TI - Managing nicotine dependence. PMID- 9207634 TI - The electrocardiogram predicts one-year outcome of patients with unstable angina and non-Q wave myocardial infarction: results of the TIMI III Registry ECG Ancillary Study. Thrombolysis in Myocardial Ischemia. AB - OBJECTIVES: We sought to determine the prognostic value of the admission electrocardiogram (ECG) in patients with unstable angina and non-Q wave myocardial infarction (MI). BACKGROUND: Although the ECG is the most widely used test for evaluating patients with unstable angina and non-Q wave MI, little prospective information is available on its value in predicting outcome in the current era of aggressive medical and interventional therapy. METHODS: ECGs with the qualifying episode of pain were analyzed in patients enrolled in the Thrombolysis in Myocardial Ischemia (TIMI) III Registry, a prospective study of patients admitted to the hospital with unstable angina or non-Q wave MI. RESULTS: New ST segment deviation > or = 1 mm was present in 14.3% of 1,416 enrolled patients, isolated T wave inversion in 21.9% and left bundle branch block (LBBB) in 9.0%. By 1-year follow-up, death or MI occurred in 11% of patients with > or = 1 mm ST segment deviation compared with 6.8% of patients with new, isolated T wave inversion and 8.2% of those with no ECG changes (p < 0.001 when comparing ST with no ST segment deviation). Two other high risk groups were identified: those with only 0.5-mm ST segment deviation and those with LBBB, whose rates of death or MI by 1 year were 16.3% and 22.9%, respectively. On multivariate analysis, ST segment deviation of either > or = 1 mm or > or = 0.5 mm remained independent predictors of death or MI by 1 year. CONCLUSIONS: The admission ECG is very useful in risk stratifying patients with non-Q wave MI. The new criteria of not only > or = 1-mm ST segment deviation but also > or = 0.5-mm ST segment deviation or LBBB identify high risk patients, whereas T wave inversion does not add to the clinical history in predicting outcome. PMID- 9207635 TI - Outcome and profile of women and men presenting with acute coronary syndromes: a report from TIMI IIIB. TIMI Investigators. Thrombolysis in Myocardial Infarction. AB - OBJECTIVES: Women and men enrolled in the Thrombolysis in Myocardial Infarction (TIMI) IIIB trial of unstable angina and non-Q wave myocardial infarction (MI) were evaluated to determine gender differences in characteristics and outcome. BACKGROUND: Coronary heart disease is the leading cause of death for women and men. However, the characteristics and outcome of women compared with men with unstable angina and non-Q wave MI have not been extensively studied. METHODS: The characteristics, outcomes and proportion of 497 women and 976 men with unstable angina and non-Q wave MI at the time of enrollment were compared. When these proportions were noted to be significantly different, we compared them with the 7,731-patient TIMI IIIB Registry, which represents the non-trial, screened population with these syndromes at these centers. RESULTS: For both coronary syndromes, women were older, were less frequently white, had a higher incidence of diabetes and hypertension and were receiving more cardiac medications. The 42 day rate of death and MI in TIMI IIIB was similar for women and men (7.4% vs. 7.5%). Coronary angiography revealed less severe coronary artery disease for women than for men, with absence of critical obstructions in 25% versus 16% and mean ejection fractions 62 +/- 12% versus 57 +/- 13% for women versus men (p < 0.01). Medical management failed in women as often as in men, and rates of cardiac catheterization and percutaneous transluminal coronary angioplasty or coronary artery bypass graft surgery were similar for women and men in the conservative strategy arm as well as in the invasive strategy arm. Women in the TIMI IIIB trial had proportionately more unstable angina than did men. The proportion of unstable angina and non-Q wave MI for women was similar in the trial and Registry. However, proportionately more men in the trial had non-Q wave MI than men in the Registry. CONCLUSIONS: 1) Women with each acute coronary syndrome are older than men and have more comorbidity. 2) The outcome with unstable angina and non-Q wave MI is related to severity of illness and not gender. 3) Mortality associated with revascularization for unstable angina and non-Q wave MI was similar for women and men. 4) The proportion of women and men enrolled with each acute coronary syndrome is different. These rates reflect both the prevalence of disease and selection bias owing to trial eligibility criteria and other identified factors. PMID- 9207636 TI - Evidence for prevention of death and myocardial infarction with platelet membrane glycoprotein IIb/IIIa receptor blockade by abciximab (c7E3 Fab) among patients with unstable angina undergoing percutaneous coronary revascularization. EPIC Investigators. Evaluation of 7E3 in Preventing Ischemic Complications. AB - OBJECTIVES: We sought to evaluate whether patients with unstable angina undergoing coronary intervention derive particular clinical benefit from potent platelet inhibition. BACKGROUND: Plaque rupture and platelet aggregation are pathogenetic processes common to unstable angina and ischemic complications of percutaneous coronary intervention. METHODS: Of the 2,099 patients undergoing a coronary intervention in the Evaluation of 7E3 in Preventing Ischemic Complications (EPIC) trial, 489 were enrolled with the diagnosis of unstable angina and randomized to receive placebo, an abciximab (c7E3) bolus immediately before the intervention or an abciximab bolus followed by a 12-h infusion. The primary end point was a composite of death, myocardial infarction (MI) or urgent repeat revascularization within 30 days of randomization. The occurrence of death, MI or any revascularization within 6 months was also assessed. RESULTS: Compared with placebo, the bolus and infusion of abciximab resulted in a 62% reduction in the rate of the primary end point (12.8% vs. 4.8%, p = 0.012) among patients with unstable angina, due primarily to a reduction in the incidences of death (3.2% vs. 1.2%, p = 0.164) and MI (9% vs. 1.8%, p = 0.004). By 6 months, cumulative death and MI were further reduced by abciximab (6.6% vs. 1.8%, p = 0.018 and 11.1% vs. 2.4%, p = 0.002, respectively). The magnitude of the risk reduction with abciximab was greater among the patients with unstable angina than among other patients in the EPIC trial without unstable angina for the end points of death (interaction: p = 0.008 at 30 days, p = 0.002 at 6 months) and MI (interaction: p = 0.004 at 30 days, p = 0.003 at 6 months). CONCLUSIONS: The syndrome of unstable angina identifies patients who will experience particularly marked reductions in the risk of death and MI with abciximab during coronary intervention. PMID- 9207637 TI - Can we provide reperfusion therapy to all unselected patients admitted with acute myocardial infarction? AB - OBJECTIVES: This study sought to assess the maximal rate of acute Thrombolysis in Myocardial Infarction (TIMI) grade 3 patency that can be achieved in unselected patients. BACKGROUND: Early and complete (TIMI grade 3 flow) reperfusion is an important therapeutic goal during acute myocardial infarction. However, thrombolysis, although widely used, is often contraindicated or ineffective. The selective use of primary and rescue percutaneous transluminal coronary angioplasty (PTCA) may increase the number of patients receiving reperfusion therapy. METHODS: A cohort of 500 consecutive unselected patients with acute myocardial infarction were prospectively treated using a patency-oriented scheme: Thrombolysis-eligible patients received thrombolysis (n = 257) and underwent 90 min angiography to detect persistent occlusion for treatment with rescue PTCA. Emergency PTCA (n = 193) was attempted in patients with contraindications to thrombolysis, cardiogenic shock or uncertain diagnosis and in a subset of patients admitted under "ideal conditions." A small group of patients (n = 38) underwent acute angiography without PTCA. Conventional medical therapy was used in 12 patients with contraindications to both thrombolysis and PTCA. RESULTS: Ninety-eight percent of patients received reperfusion therapy (thrombolysis, PTCA or acute angiography), and angiographically proven early TIMI grade 3 patency was achieved in 78%. Among patients with TIMI grade 3 patency, thrombolysis alone was the strategy used in 37%, emergency PTCA in 40% and rescue PTCA after failed thrombolysis in 15%; spontaneous patency occurred in 8%. CONCLUSIONS: Reperfusion therapy can be provided to nearly every patient (98%) with acute myocardial infarction. Rescue and direct PTCA provided effective early reperfusion to patients in whom thrombolysis failed or was excluded. PMID- 9207638 TI - Induction of cytokine expression in leukocytes in acute myocardial infarction. AB - OBJECTIVES: This study sought to investigate whether cytokine expression in leukocytes may be induced by plasma from the reperfused heart of patients with an acute myocardial infarction (MI). BACKGROUND: Reperfusion in acute MI is associated with deleterious local and systemic inflammatory responses that are regulated by cytokines. Induction of cytokine expression in resident leukocytes could contribute to inflammatory responses of the ischemic and reperfused heart. METHODS: Blood samples of 10 patients with an acute MI were obtained simultaneously from the coronary sinus and the aorta before and 5 min after recanalization of the coronary occlusion. Ten patients with elective percutaneous transluminal coronary angioplasty served as a control group. We incubated leukocytes from healthy donors with plasma samples and analyzed mRNA expression of interleukin (IL)-1 beta, IL-6, IL-8 and tumor necrosis factor-alpha (TNF alpha) by Northern blot analysis. RESULTS: In patients with an acute MI, plasma obtained from the coronary sinus after recanalization increased the mRNA expression of IL-1 beta and IL-8 compared with that of plasma before recanalization (median [quartiles] difference before vs. after recanalization: 34.5 [4, 137], p = 0.017, for IL-1 beta; 18.5 [4, 35], p = 0.032, for IL-8) and simultaneously obtained aortic plasma (median [quartiles] coronary sinus-aortic differences after recanalization: 45.5 [-3, 115], p = 0.021, for IL-1 beta; 16 [4, 52], p = 0.005, for IL-8). No induction of IL-6 and TNF-alpha expression could be observed. No changes found in the study patients were detectable in the control group. CONCLUSIONS: Plasma from the ischemic and reperfused heart stimulates the expression of IL-1 beta and IL-8 in leukocytes. Therefore, leukocyte-derived cytokines may contribute to the regulation of cardiac inflammatory responses in patients with an acute MI. PMID- 9207640 TI - Multivessel Palmaz-Schatz stenting: early results and one-year outcome. AB - OBJECTIVES: To determine whether the benefits outlined in Background might extend to patients with multivessel disease, we examined the short- and long-term outcome of multivessel Palmaz-Schatz stenting. BACKGROUND: Percutaneous transluminal coronary angioplasty (PTCA) has become the dominant treatment for most patients with single-vessel coronary artery disease and has emerged as an alternative treatment for selected patients with multivessel coronary artery disease. Although multivessel angioplasty has excellent early results and low procedural complication rates, long-term outcome is tempered by the frequent need for repeat revascularization. In patients with single-vessel coronary artery disease, Palmaz-Schatz stenting has been shown to have a higher success rate and a lower restenosis rate than conventional PTCA. METHODS: A total of 103 patients (mean age 64 +/- 11 years, 78 men and 25 women) underwent stenting of 212 vessels (saphenous vein graft [53%], left anterior descending coronary artery [20%], left circumflex artery [12%] and right coronary artery [15%]). In 88 patients (85%), multivessel stenting was performed during the same procedure, whereas the remaining 15 patients (15%) had staged multivessel stenting within 1 week of the index stent. Stenting involved only native coronary arteries in 33 patients and only vein grafts in 51 patients. RESULTS: Angiographic success was achieved in 102 patients (99%). Major complications developed in three patients: one patient died, and two patients had Q wave myocardial infarction, with no emergency coronary artery bypass graft surgery or stent thrombosis. Eleven additional patients (11%) developed non-Q wave myocardial infarction, and nine patients (9%) had local vascular complications requiring surgical repair. Clinical follow-up was available in all patients at a mean of 13 +/- 8 months. At 1 year, survival was 98%, with an event-free survival rate of 80%, reflecting predominantly repeat revascularization (17% overall, with 9% target site revascularization). Multivessel native coronary stenting resulted in a higher event-free survival rate and a lower probability of repeat revascularization than did multivessel saphenous vein graft stenting. CONCLUSIONS: In selected patients, multivessel Palmaz-Schatz stenting is technically feasible and carries both excellent early results and favorable 1-year clinical outcome. PMID- 9207639 TI - Influence of diabetes mellitus on clinical outcome in the thrombolytic era of acute myocardial infarction. GUSTO-I Investigators. Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries. AB - OBJECTIVES: This study was undertaken to define and better understand the characteristics and outcomes of patients with diabetes treated for acute myocardial infarction with contemporary thrombolysis. BACKGROUND: Although thrombolysis has substantially improved survival of patients with myocardial infarction, diabetes mellitus remains an independent predictor for a poor prognosis. METHODS: We characterized the contemporary relation between diabetes and outcome after myocardial infarction treated with thrombolytic agents from a large international cohort. Of 41,021 patients randomized to receive accelerated tissue-type plasminogen activator (t-PA), streptokinase or a combination of both agents in the Global Utilization of Streptokinase and Tissue Plasminogen Activator for Occluded Coronary Arteries study, there were 5,944 patients with diabetes and 34,888 patients without diabetes. RESULTS: Patients with diabetes were older and more likely to be female, to present with anterior wall infarction, to receive thrombolysis later and to have triple-vessel coronary artery disease. Mortality at 30 days was highest among diabetic patients treated with insulin (12.5%) compared with non-insulin-treated diabetic (9.7%) and nondiabetic (6.2%) patients (p < 0.001). Mortality was lowest among those with diabetes receiving accelerated t-PA, which is consistent with the results of the overall patient cohort. Although stroke occurred more frequently among diabetic (1.9%) than nondiabetic patients (1.4%, p < 0.001), there was no significant difference in the rates of intracranial hemorrhage. Cardiac failure, shock, atrioventricular block and atrial flutter/ fibrillation were more common among diabetic patients. The proportion of patients undergoing revascularization was similar between patients with and without diabetes, although diabetic patients were more likely to undergo coronary artery bypass graft surgery (10.4% vs. 8.3%). Diabetes remained an independent predictor for mortality at 1-year follow up (14.5% vs. 8.9%, p < 0.001). CONCLUSIONS: Diabetes, alone and in association with its comorbidities, portends a substantially worse 30-day and 1-year prognosis for patients with myocardial infarction. PMID- 9207641 TI - Long-term clinical follow-up after successful repeat percutaneous intervention for stent restenosis. AB - OBJECTIVES: This study evaluated the long-term clinical outcome of successful repeat percutaneous intervention after in-stent restenosis. BACKGROUND: Recurrence of symptoms and angiographic restenosis after stent implantation are observed in 15% to 35% of cases. Repeat percutaneous treatment for in-stent restenosis has been shown to be safe, with high immediate success, but little is known about the long-term clinical outcome. METHODS: Clinical follow-up (minimum 9 months) was obtained in a consecutive series of 124 patients (127 vessels) presenting with stent restenosis who were successfully treated with repeat percutaneous intervention. RESULTS: Clinical follow-up was obtained in all 124 patients at a mean [+/-SD] of 27.4 +/- 14.7 months (range 9 to 66); a stress test was available in 88 patients (71%). Recurrence of clinical events occurred in 25 patients (20%) and included death from any cause in 2 patients (2%), target vessel revascularization in 14 (11%), myocardial infarction in 1 (1%) and positive stress test results or recurrence of symptoms (Canadian Cardiovascular Society class I to IV) treated medically in 8 (6%). Cumulative event-free survival at 12 and 24 months was 86.2% and 80.7%, respectively. Significant predictive factors of recurrence of clinical events were repeat intervention in saphenous vein grafts, multivessel disease, low ejection fraction and a < or = 3 month interval between stent implantation and repeat intervention. CONCLUSIONS: In-stent balloon angioplasty for stent restenosis in native vessels seems to be an effective method in terms of a low long-term clinical event rate. PMID- 9207642 TI - Risk of major complications from coronary angioplasty performed immediately after diagnostic coronary angiography: results from the Registry of the Society for Cardiac Angiography and Interventions. AB - OBJECTIVES: This study was designed to determine the risk of performing percutaneous transluminal coronary angioplasty (PTCA) at the time of diagnostic catheterization ("combined procedures"). BACKGROUND: Health care providers are under increasing pressure to combine diagnostic and interventional coronary procedures to reduce costs. However, the risk associated with combined procedures has not been rigorously assessed. METHODS: A multicenter cohort study of 35,700 patients undergoing elective PTCA from 1992 through 1995 was performed to determine the risk of major complications (myocardial infarction, emergency coronary artery bypass graft surgery or death) from combined relative to staged procedures (i.e., performing PTCA at a session subsequent to diagnostic catheterization). RESULTS: The risks of major complications from combined and staged procedures were 2.0% and 1.6%, respectively (unadjusted odds ratio [OR] 1.28, 95% confidence interval [CI] 1.05 to 1.57). After adjusting for clinical and angiographic differences and clustering by laboratory, the risk from combined procedures was not significantly elevated (multivariable OR 1.18, 95% CI 0.89 to 1.55). However, several subgroups of patients did have an increased risk from combined procedures: patients with multivessel disease (multivariable OR 1.64, 95% CI 1.13 to 2.39); women (multivariable OR 1.64, 95% CI 1.05 to 2.55); patients > 65 years old (multivariable OR 1.40, 5% CI 1.02 to 1.93); and patients undergoing multilesion PTCA (multivariable OR 1.53, 95% CI 1.06 to 2.21). The risk of combined relative to staged procedures decreased over the 4-year period (multivariable p = 0.029). CONCLUSIONS: Combining PTCA with diagnostic catheterization appears to be safe in many patients. However, several subgroups of patients may be at increased risk. Careful patient selection will most likely remain critical to ensuring the safety of combined procedures. PMID- 9207643 TI - Predictors of success and major complications for primary percutaneous transluminal coronary angioplasty in acute myocardial infarction. An analysis of the 1990 to 1994 Society for Cardiac Angiography and Interventions registries. AB - OBJECTIVES: The purpose of this study was to determine predictors of successful coronary angioplasty for acute myocardial infarction (MI) and associated predictors of the major complications of in-hospital mortality and emergency coronary artery bypass graft surgery. BACKGROUND: Primary angioplasty is being increasingly used to treat acute MI, but factors affecting the success and major complications have not been well studied. Forty laboratories have been contributing clinical and procedural data to the Society of Cardiac Angiography and Interventions (SCA&I) on primary angioplasty for acute MI. METHODS: Univariable and stepwise multivariable logistic regression analysis of clinical and procedural variables was used to calculate predictors of success and major complications. RESULTS: There were 4,366 primary angioplasty procedures reported from 1990 through 1994, with an overall success rate of 91.5%, an in-hospital mortality rate of 2.5% and a rate of emergency surgery of 4.3%. Higher laboratory primary angioplasty volume and lower age were predictive of success. An intraaortic balloon pump in place, cardiogenic shock and a moribund condition had negative predictive effects. Unsuccessful angioplasty, cardiogenic shock or a moribund state were predictive of in-hospital death. Unsuccessful angioplasty, the absence of a history of hypertension and the absence of congestive heart failure were predictive of emergency surgery. CONCLUSIONS: The rates of success and major complications in the SCA&I Registry are similar to other series. Predictors of success and major complications can be assessed and may be useful for risk stratifying candidates for primary angioplasty in acute MI. PMID- 9207644 TI - Time- and rate-dependent alterations of the QT interval precede the onset of torsade de pointes in patients with acquired QT prolongation. AB - OBJECTIVES: The purpose of this study was to determine whether the QT interval dynamics that precede torsade de pointes are consistent with the initiation of this arrhythmia by early afterdepolarization-induced triggered activity. BACKGROUND: Early afterdepolarization-induced triggered activity has been suggested as an electrophysiologic mechanism for torsade de pointes. Consequently, the initiation of torsade de pointes should involve time- and rate dependent alterations of ventricular repolarization similar to those known to modulate the development of early afterdepolarizations. METHODS: RR and QT intervals were measured in digitized 24-h ambulatory electrocardiographic recordings obtained from seven patients with acquired prolongation of ventricular repolarization. Each patient had one or more episodes of torsade de pointes. The relation between RR and QT intervals was determined before, during and after multiple episodes of torsade de pointes. RESULTS: In patients with multiple episodes of ventricular arrhythmias, the onset of the arrhythmias was associated with a critical prolongation of the QT interval. In some episodes, prolongation of the QT interval was associated with sudden prolongation of the sinus cycle length, whereas in other episodes, the QT interval prolonged progressively at a constant cycle length. CONCLUSIONS: The association between a critically prolonged QT interval and the onset of ventricular arrhythmias suggests that the initial complex of torsade de pointes is an early afterdepolarization-induced triggered response. However, prolongation of the QT interval itself was not sufficient to account for the initiation of torsade de pointes, suggesting that other, as yet unidentified factors are required. PMID- 9207645 TI - Atypical atrioventricular node reciprocating tachycardia masquerading as tachycardia using a left-sided accessory pathway. AB - OBJECTIVES: The study was performed to document that atrioventricular node reciprocating tachycardia (AVNRT) can be associated with eccentric retrograde left-sided activation, masquerading as tachycardia using a left accessory pathway. BACKGROUND: The eccentric retrograde left-sided activation during tachycardia is thought to be diagnostic of the presence of a left free wall accessory pathway. However, it is not known whether AVNRT can occur with eccentric retrograde left-sided activation. METHODS: We studied 356 patients with AVNRT who underwent catheter ablation. Retrograde atrial activation during tachycardia and ventricular pacing were determined by intracardiac recordings, including the use of a decapolar coronary sinus catheter. RESULTS: The retrograde atrial activation was eccentric in 20 patients (6%). Eight of these patients had the earliest retrograde atrial activation recorded in the lateral coronary sinus leads, and 12 had the earliest retrograde atrial activation recorded in the posterior coronary sinus leads, with the most proximal coronary sinus electrode pair straddling the coronary sinus orifice. These tachycardias were either the fast-slow or the slow-slow form of AVNRT. The slow-fast form of AVNRT was also inducible in 17 of the 20 patients. Successful ablation of the slow pathway in the right atrial septum near the coronary sinus ostium prevented the induction and clinical recurrence of reciprocating tachycardia in all patients. CONCLUSIONS: Atypical AVNRT with eccentric retrograde left-sided activation was demonstrated in 6% of all patients with AVNRT masquerading as tachycardia using a left-sided accessory pathway. Ablation of the slow pathway at the posterior aspects of the right atrial septum resulted in a cure in these patients. PMID- 9207646 TI - Predictive value for future arrhythmic events of fractal dimension, a measure of time clustering of ventricular premature complexes, after myocardial infarction. CAST Investigators. Cardia Arrhythmia Suppression Trial. AB - OBJECTIVES: Our objective was to test fractal dimension (D), a measure of clustering of ventricular premature complexes (VPCs), on entry Holter recording as a predictor of future arrhythmic death and other-cause mortality in postinfarction patients in the Cardiac Arrhythmic Suppression Trial (CAST). BACKGROUND: Nonlinear dynamic methods of signal processing are being applied in medicine to provide new insights into apparently "chaotic" biologic events, including cardiac arrhythmias. One such application is the derivation of a fractal D to describe the clustering of VPCs in time. METHODS: Baseline Holter recordings were analyzed in blinded manner for 484 patients: 237 died or had a resuscitated cardiac arrest during follow-up, and 247 matched patients had no events. Fractal D, measured in four ways, was assessed as a predictor using Cox regression. RESULTS: One measure of D (high resolution D) was a significant univariate (relative hazard ratio 0.79 per SD change, p = 0.011) and multivariate (hazard ratio 0.75, p = 0.046) predictor of arrhythmic death but not other death (univariate p = 0.95, relative hazard 0.95, p = 0.66). Fractal D was greater (VPCs less clustered) in those patients free of arrhythmic events. On subgroup analysis, the predictive value of D resided in the randomized patient group (i.e., those who showed VPC suppression during initial antiarrhythmic drug titration and were randomized to blinded therapy with active drug or placebo) (multivariate hazard ratio 0.57, p = 0.001). CONCLUSIONS: A high resolution fractal D was predictive of arrhythmic (but not nonarrhythmic) death in a large postinfarction cohort. Further study of this new signal processing approach to ambulatory electrocardiographic recording will be of interest. PMID- 9207647 TI - Biphasic waveforms prevent the chronic rise of defibrillation thresholds with a transvenous lead system. AB - OBJECTIVES: The purpose of this study was to compare chronic changes in monophasic and biphasic defibrillation thresholds using a uniform transvenous lead system and testing protocol. BACKGROUND: Defibrillation thresholds increase over time in patients with nonthoracotomy lead systems. This increase can result in an inadequate chronic defibrillation safety margin and could limit the safety of smaller pulse generators, which have a reduced maximal output. However, previous studies of the temporal changes of defibrillation thresholds evaluated complex lead systems or monophasic shock waveforms, neither of which are used with current technology. METHODS: This study was a prospective, randomized assessment of the effects of shock waveforms on the changes of transvenous defibrillation thresholds over time. Paired monophasic and biphasic thresholds were measured both at implantation and at follow-up (250 +/- 105 days) in 24 consecutive patients who were not receiving antiarrhythmic drugs. The lead system was a dual-coil Endotak C lead, and reverse polarity shocks (distal coil = anode) were delivered. RESULTS: Monophasic defibrillation thresholds increased from (mean +/- SD) 13.7 +/- 6.0 J to 16.8 +/- 6.7 J (p = 0.02), whereas biphasic thresholds were unchanged (10.4 +/- 4.3 J to 10.2 +/- 4.8 J, p = 0.86) in the same patients. Shock impedance chronically increased (47.0 omega to 50.5 omega, p = 0.02) and was unaffected by waveform. CONCLUSIONS: These results indicate that biphasic shocks prevent the chronic increase in defibrillation thresholds with a transvenous lead system. PMID- 9207648 TI - Hemodynamic and clinical effects of oral levodopa in children with congestive heart failure. AB - OBJECTIVES: This study was undertaken to evaluate the safety, efficacy and pharmacodynamic variables of oral levodopa in pediatric patients with congestive heart failure refractory to standard therapy. BACKGROUND: Therapeutic options for children with congestive cardiomyopathies are limited to digoxin, diuretic agents and angiotensin-converting enzyme inhibitors. Previous work in adults with congestive heart failure has shown a short-term effectiveness of levodopa and improvement of cardiac function. METHODS: Baseline two-dimensional and M-mode echocardiography, surface electrocardiography, Holter monitoring and exercise testing, when applicable, were performed. Levodopa was administered in a dose escalation scale from 8 mg/kg body weight per dose to 20 mg/kg per dose over 3 days with concomitant metoclopramide and pyridoxine. Catecholamine levels at initiation of the trial and throughout dose escalation were measured, with echocardiographic and electrocardiographic correlation. After 24-h drug washout, cardiac catheterization was performed both before and after administration of levodopa. RESULTS: Between February 1992 and December 1995, nine children (age 10 +/- 1.7 years, weight 27.8 +/- 4.3 kg) were enrolled in this study. At cardiac catheterization, serum dopamine levels rose from 108.5 +/- 59.2 pg/ml to 1,375.8 +/- 567.9 pg/ml (p = 0.03) at 100 +/- 14.8 min after levodopa administration without a significant change in serum norepinephrine or epinephrine levels. Paralleling these increases, there were significant changes in the cardiac index (1.7 +/- 0.3 to 3.2 +/- 0.7 liters/min per m2), stroke volume index (16.1 +/- 3.2 to 31.2 +/- 7.0 ml/m2 per min), oxygen consumption (138.6 +/- 24.4 to 188.3 +/- 30.8 ml/min per m2) and systemic vascular resistance (36.8 +/- 8 to 21.9 +/- 5.5 indexed Wood's units; all p < 0.01). There was a significant reversal of the daily fluid volume output/input ratio from 0.8 +/- 0.1 to 1.2 +/- 0.1 (p < 0.01). Levodopa administration was complicated by hypertension or tachycardia, or both, requiring a dose reduction in three patients, and by significant gastrointestinal distress in one. There was sustained symptomatic improvement a median of 19.5 months after drug initiation in seven of the patients. CONCLUSIONS: These preliminary data support the hemodynamic value of oral levodopa in the treatment of severe congestive heart failure in children. PMID- 9207649 TI - Children with heart murmurs: can ventricular septal defect be diagnosed reliably without an echocardiogram? AB - OBJECTIVES: This study was undertaken to determine the accuracy of expert examination for ventricular septal defect (VSD) among children with a heart murmur. BACKGROUND: Because the frequency and nature of errors that might be made by reliance solely on expert examination for diagnosis of VSD are speculative, the role of echocardiography in such diagnosis is controversial. METHODS: Two hundred eighty-seven consecutive previously unevaluated pediatric subjects were enrolled in the study. For each child, the pediatric cardiologists prospectively recorded a working diagnosis and their level of confidence in the diagnosis, categorizing any VSD diagnosed as small or moderate to large. After echocardiography, VSDs were subcategorized by location and requirement for treatment as minor, intermediate or major. Receiver-operating characteristic (ROC) curves described the accuracy of the clinical examination. RESULTS: Seventy three subjects had a VSD (minor in 52, intermediate in 10 and major in 11). ROC areas (1.0 = perfect discrimination, 0.5 = indiscriminate) were minor VSD 0.92 +/ 0.02 and major/intermediate VSD 0.69 +/- 0.07 (p = 0.0016). Four of 52 minor VSDs were not identified at any level of suspicion; the clinical diagnoses were moderate to large VSD in two patients and atrial septal defect and unlimited differential diagnosis in one patient each. Fourteen of 235 patients without a minor VSD were believed with confidence to have a small VSD, but the final diagnosis was intermediate VSD in 4, innocent murmur in 3, major VSD in 2, pulmonary stenosis in 2 and subaortic membrane, atrial septal defect and mitral regurgitation in 1 patient each. CONCLUSIONS: Almost all minor VSDs are recognized without echocardiography; however, errors can occur even when an expert examiner is confident. Clinical recognition of an intermediate or major VSD is less accurate than clinical recognition of a minor VSD. Failure to distinguish VSDs of major or intermediate importance from minor VSDs is a weakness of the expert clinical examination. PMID- 9207650 TI - Potential role of mechanical stress in the etiology of pediatric heart disease: septal shear stress in subaortic stenosis. AB - OBJECTIVES: The objective of this study was to show elevations in septal shear stress in response to morphologic abnormalities that have been associated with discrete subaortic stenosis (SAS) in children. Combined with the published data, this critical connection supports a four-stage etiology of SAS that is advanced in this report. BACKGROUND: Subaortic stenosis constitutes up to 20% of left ventricular outflow obstruction in children and frequently requires surgical removal, and the lesions may reappear unpredictably after the operation. The etiology of SAS is unknown. This study proposes a four-stage etiology for SAS that I) combines morphologic abnormalities, II) elevation of septal shear stress, III) genetic predisposition and IV) cellular proliferation in response to shear stress. METHODS: Morphologic structures of a left ventricular outflow tract were modeled based on measurements in patients with and without SAS. Septal shear stress was studied in response to changes in aortoseptal angle (AoSA) (120 degrees to 150 degrees), outflow tract convergence angle (45 degrees, 22.5 degrees and 0 degree), presence/location of a ventricular septal defect (VSD) (3 mm VSD; 2 and 6 mm from annulus) and shunt velocity (3 and 5 m/s). RESULTS: Variations in AoSA produced marked elevations in septal shear stress (from 103 dynes/cm2 for 150 degrees angle to 150 dynes/cm2 for 120 degrees angle for baseline conditions). This effect was not dependent on the convergence angle in the outflow tract (150 to 132 dynes/cm2 over full range of angles including extreme case of 0 degree). A VSD enhanced this effect (150 to 220 dynes/cm2 at steep angle of 120 degrees and 3 m/s shunt velocity), consistent with the high incidence of VSDs in patients with SAS. The position of the VSD was also important, with a reduction of the distance between the VSD and the aortic annulus causing further increases in septal shear stress (220 and 266 dynes/cm2 for distances of 6 and 2 mm from the annulus, respectively). CONCLUSIONS: Small changes in AoSA produce important changes in septal shear stress. The levels of stress increase are consistent with cellular flow studies showing stimulation of growth factors and cellular proliferation. Steepened AoSA may be a risk factor for the development of SAS. Evidence exists for all four stages of the proposed etiology of SAS. PMID- 9207651 TI - Abnormalities of the left ventricular outflow tract associated with discrete subaortic stenosis in children: an echocardiographic study. AB - OBJECTIVES: The purpose of this study was to examine the echocardiographic abnormalities of the left ventricular outflow tract associated with subaortic stenosis in children. BACKGROUND: Considerable evidence suggests that subaortic stenosis is an acquired and progressive lesion, but the etiology remains unknown. We have proposed a four-stage etiologic process for the development of subaortic stenosis. This report addresses the first stage by defining the morphologic abnormalities of the left ventricular outflow tract present in patients who develop subaortic stenosis. METHODS: Two study groups were evaluated-33 patients with isolated subaortic stenosis and 12 patients with perimembranous ventricular septal defect and subaortic stenosis-and were compared with a size- and lesion matched control group. Subjects ranged in age from 0.05 to 23 years, and body surface area ranged from 0.17 to 2.3 m2. Two independent observers measured aortoseptal angle, aortic annulus diameter and mitral-aortic separation from previously recorded echocardiographic studies. RESULTS: The aortoseptal angle was steeper in patients with isolated subaortic stenosis than in control subjects (p < 0.001). This pattern was also true for patients with ventricular septal defect and subaortic stenosis compared with control subjects (p < 0.001). Neither age nor body surface area was correlated with aortoseptal angle. A trend toward smaller aortic annulus diameter indexed to patient size was seen between patients and control subjects but failed to achieve statistical significance (p = 0.08). There was an excellent interrater correlation in aortoseptal angle and aortic annulus measurement. The mitral-aortic separation measurement was unreliable. Our results, specifically relating steep aortoseptal angle to subaortic stenosis, confirm the results of other investigators. CONCLUSIONS: This study demonstrates that subaortic stenosis is associated with a steepened aortoseptal angle, as defined by two-dimensional echocardiography, and this association holds in patients with and without a ventricular septal defect. A steepened aortoseptal angle may be a risk factor for the development of subaortic stenosis. PMID- 9207652 TI - ACC/AHA Guidelines for Exercise Testing. A report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Committee on Exercise Testing). PMID- 9207653 TI - Resorbable plate fixation in pediatric craniofacial surgery. AB - Resorbable bone plates composed of a copolymer of polylactic and polyglycolic acids stabilized into position with metallic microscrews were used in the reconstruction of pediatric craniofacial deformities. In 100 patients between 4 and 15 months of age, a total of 912 resorbable plates were implanted over a 2 1/2-year period. Their application was simple and rapid and required no special instrumentation. Currently, 85 patients are more than 1 year postimplantation, which is the known time for complete resorption of this copolymeric compound. No complications have been seen with this use, including infection, overlying soft tissue reactions, reconstructive instability, or underlying osteolysis around the screws, as determined by postoperative plain radiographs at 6 months and 1 year postoperative time periods. Four patients have had screws removed due to either palpability or secondary reconstructive surgery between 9 and 18 months postoperatively, all of whom exhibited complete polymer resorption and normal bone healing. These clinical results demonstrate the safety and effectiveness of this copolymeric material for pediatric craniofacial applications. PMID- 9207654 TI - Biocompatibility of fixation materials in the brain. AB - Recent clinical reports documenting passive intracranial translocation of microplates and microscrews have prompted concerns regarding brain biocompatibility and neurotoxicity of fixation hardware used in craniofacial surgery. Although the effects of commercially pure titanium. Vitallium (cobalt chromium-molybdenum), stainless steel, and various alloys have been well assessed in bone and soft tissues, there are no comprehensive studies of these materials in the brain. To investigate this, the biocompatibility of titanium, vitallium, and 316L stainless steel was evaluated in the rabbit brain and compared with silicone elastomer shunt tubing, a material that is used commonly as a neurosurgical implant material with well-established brain biocompatibility. Forty-eight juvenile New Zealand White rabbits were randomly assigned to one of three groups and underwent placement of either commercially pure titanium microscrews, vitallium microscrews, or 316L monofilament stainless steel wire into the parietal region. Silicone elastomer strips of similar size were implanted in the contralateral hemisphere of each rabbit. Animals were assessed daily for signs of neurotoxicity. Rabbits in each group were sacrificed at 2, 4, 8, and 26 weeks following implantation. Brains were sectioned at the implantation site and were examined by means of standard hematoxylin and eosin stains and with immunohistochemical markers sensitive to inflammatory changes in the brain. None of the animals showed any behavioral changes or neurologic deficits suggestive of either systemic or localized toxicity from the implant materials. Silicone clastomer was found to cause the least amount of inflammation relative to other materials tested at all sacrifice points, suggesting that as a standard neurosurgical implant material, it is an appropriate control for studies of brain biocompatibility. At 2 weeks, titanium was found to cause the largest inflammatory response in surrounding brain parenchyma based on analysis of markers for microglial proliferation, gliosis, and leukocyte infiltration. At the 26-week endpoint of our study, the biocompatibility of titanium was nearly equal to the biocompatibility of vitallium based on all studied markers of inflammation. A progressive increase in the inflammatory response surrounding stainless steel implants was noted at 8 and 26 weeks. Relative to all materials studied, at 26 weeks the greatest leukocyte response was found with stainless steel implants. Our results indicate that at the 26-week end-point of our study, titanium and vitallium incited a similar inflammatory response in the rabbit brain that was greater than the response found with silicone elastomer, a standard neurosurgical implant material, but less than that found with stainless steel wire, which is commonly recommended as an alternative fixation material. PMID- 9207655 TI - Regional differences of dura osteoinduction: squamous dura induces osteogenesis, sutural dura induces chondrogenesis and osteogenesis. AB - Dura plays an important role in calvarial morphogenesis. However, precisely what that role is remains unclear. We present here in vivo evidence that dura without other central nervous system components induces both chondrogenesis and osteogenesis. The mechanism is, at least in part, by proximate tissue interaction. The objectives of this experiment were to answer the following: (1) Can dura actually induce osteogenesis without the influence of the underlying brain? (2) What are the requirements of this dura-induced heterotopic osteogenesis? (3) What are the differences between dura underlying sutures and dura underlying the squamous portions of the cranial bones? Dura underlying the metopic, sagittal, and lambdoidal sutures and dura underlying the flat portions of frontal and parietal bones were obtained from neonatal Lewis rats and transplanted into the posterior thoraces of adult Lewis recipients. In group I, dura underlying the metopic, sagittal, and lambdoidal sutures (n = 20) and dura underlying the flat portions of frontal and parietal bones (n = 20) were transplanted individually into separate epitheliomesenchymal pockets. Group II animals had dura underlying the metopic, sagittal, and lambdoidal sutures (n = 10) and dura underlying the flat portions of frontal and parietal bones (n = 10) transplanted individually into surgically created mesenchymal pockets by placing the dura grafts between panniculus carnosus and latissimus dorsi muscles. The animals were sacrificed at 2-week intervals. Light microscopy, special histochemical analysis, immunohistochemistry, and electron microscopy were performed. Bone formation was seen in 15 of the 18 animals (83 percent) in group I. No bone or cartilage formation was seen in group II. Chondrogenesis was seen in 4 animals receiving dura underlying the metopic, sagittal, and lambdoidal sutures in group I. Cellular hyperproliferation was seen at 2 weeks when dura was transplanted close to the hair follicles. These cells had a high nucleus-to cytoplasm ratio and were positive for transforming growth factor beta. This hyperproliferation was followed by production and accumulation of Alcian blue positive extracellular matrix that resisted digestion by hyaluronidase. Cellularly active cartilage was seen at 6 weeks. There was no chondrogenesis in animals receiving dura underlying the flat portions of frontal and parietal bones in group I. Electron microscopy demonstrated the presence of proteoglycan-like ground substance and type II collagen in the inner layer of sutural dura and the predominance of dense type I collagen in the squamous dura and the external layer of the sutural dura. The important findings of this experiment are that (1) heterotopically transplanted neonatal dura can induce osteogenesis, (2) this heterotopic osteoinduction by dura requires epitheliomesenchymal interaction, and (3) separating dura into sutural dura and squamous dura, chondrogenesis occasionally occurred in addition to osteogenesis with the former, while only membranous ossification occurred with the latter, indicating intrinsic differences within the dura mater. This dural heterogeneity is supported by direct ultrastructural data. PMID- 9207656 TI - Cervical chrondrocutaneous branchial remnants. AB - Similar in appearance to preauricular tags but located in the lateral neck, cervical chondrocutaneous branchial remnants are a rather less common and less well known congenital lesion. A retrospective review of admissions at Sainte Justine Hospital between 1980 and 1993 produced 20 cases of cervical tags, of which 17 were true cervical chrondrocutaneous branchial remnants and 3 were skin tags associated with a thyroglossal duct. Of the 17 true cervical chrondrocutaneous branchial remnants, 15 were operated on in our institution. The clinical characteristics, results of investigations, surgical data, pathologic findings, and associated anomalies were documented. Several interesting facts emerged, including a male predominance (11 of 17), a scarcity of bilateral lesions (1 of 17), the presence of an elastic cartilage core in all operated specimens (15 of 15), and a high incidence of associated anomalies (13 of 17). We suggest that the second branchial arch is the most likely origin for the lesion. We propose a clear, widely acceptable name for this anomaly in order to prevent further diagnostic confusion. Most important, although simple surgical excision is all that is required for treatment, a complete physical examination of the patient and possibly an ultrasound examination of the genitourinary tract are recommended because a cervical chrondrocutaneous branchial remnant has proven in many cases to be a visible "marker" for more serious associated anomalies. PMID- 9207657 TI - The "triple technique" for treating stable Graves' ophthalmopathy. AB - Graves' ophthalmopathy may range from mild eyelid retraction to a devastating process that involves the entire orbit and culminates in gross ocular congestion, massive proptosis, and even blindness. Whether the ophthalmopathy is mild or severe, patients are managed on an individual basis according to the predominant clinical findings, which may include congestion, myopathy, lid retraction, proptosis, and optic neuropathy. The process usually becomes quiescent after 6 months to 3 years; however, the changes caused by fibrosis (lid retraction and ocular muscle enlargement) are permanent. The cornerstone of surgical treatment for severe cases is bony orbital decompression; however, in our experience, mild to moderate Graves' ophthalmopathy is better treated by combining eyelid surgery and orbital lipectomy. Our approach consists of a conservative orbital lipectomy, the lengthening of the levator-Muller complex by means of marginal myotomies, and a limited lateral tarsal apposition. These three different surgical steps, which have been described previously as isolated procedures, are undertaken on both eyes at the same time and modulated according to the deformity of the patient. The operation can be performed under local anesthesia with sedation, thus allowing intraoperative monitoring of the correction; the patient can be discharged after a few hours. The results in 32 operated eyes of 16 patients have been a marked aesthetic and functional improvement, with no complications after 6 to 18 months of follow-up. The relative simplicity and very low morbidity of this procedure, as well as its reliability, make it ideal in patients with mild to moderate aesthetic and functional impairment who are looking for a substantial improvement but are unwilling to undergo a relatively major procedure such as a transosseous decompression, which, in our opinion, is the operation of choice only when the patient presents with optic neuropathy or major proptosis. PMID- 9207658 TI - Physician rewards for different kinds of service: the RBRVS versus the CPR system. AB - The resource-based relative value scale (RBRVS) was introduced in 1992 by Medicare for payments to physicians. This replaced the previous system based on the physician's customary, prevailing, and reasonable (CPR) charges. This paper analyzes the RBRVS from two perspectives: (1) the economic logic of the system and (2) how it functions differently from the CPR system in practice. As a social pricing system, it can make sense under certain conditions. However, when we provided a test for a New York plastic surgeon of the alleged underpricing of evaluative relative to procedural services under CPR, we found evidence to the contrary. PMID- 9207659 TI - Acute ischemic preconditioning of skeletal muscle prior to flap elevation augments muscle-flap survival. AB - Ischemic preconditioning of the myocardium with repeated brief periods of ischemia and reperfusion prior to prolonged ischemia significantly reduces subsequent myocardial infarction. Following ischemic preconditioning, two "windows of opportunity" (early and late) exist, during which time prolonged ischemia can occur with reduced infarction size. The early window occurs at approximately 4 hours and the late window at 24 hours following ischemic preconditioning of the myocardium. We investigated if ischemic preconditioning of skeletal muscle prior to flap creation improved subsequent flap survival and perfusion immediately or 24 hours following ischemic preconditioning. Currently, no data exist on the utilization of ischemic preconditioning in this fashion. The animal model used was the latissimus dorsi muscle of adult male Sprague-Dawley rats. Animals were assigned to three groups, and the right or left latissimus dorsi muscle was chosen randomly in each animal. Group 1 (n = 12) was the control group, in which the entire latissimus dorsi muscle was elevated acutely without ischemic preconditioning. Group 2 (n = 8) investigated the effects of ischemic preconditioning in the early window. In this group, the latissimus dorsi muscle was elevated immediately following preconditioning. Group 3 (n = 8) investigated the effects of ischemic preconditioning in the late window, with elevation of the latissimus dorsi muscle 24 hours following ischemic preconditioning. The preconditioning regimen used in groups 2 and 3 was two 30-minute episodes of normothermic global ischemia with intervening 10-minute episodes of reperfusion. Latissimus dorsi muscle ischemia was created by occlusion of the thoracodorsal artery and vein and the intercostal perforators, after isolation of the muscle on these vessels. Muscle perfusion was assessed by a laser-Doppler perfusion imager. One week after flap elevation, muscle necrosis was quantified in all groups by means of computer-assisted digital planimetry. Our results show that ischemic preconditioning resulted in a significant reduction (p < 0.05) in muscle-flap necrosis immediately and 24 hours following ischemic preconditioning. Perfusion changes after flap elevation were similar among the three groups. Ischemic preconditioning of skeletal muscle prior to flap creation significantly reduces subsequent muscle-flap necrosis caused by the ischemia of flap creation immediately and 24 hours following ischemic preconditioning. Further elaboration of the mechanisms of ischemic preconditioning may allow pharmacologic preconditioning to be used in the augmentation of skeletal muscle-flap survival in the clinical setting. PMID- 9207660 TI - The effect of pentoxifylline on random-pattern skin-flap necrosis induced by nicotine treatment in the rat. AB - Cigarette smoke, and specifically nicotine, has been shown to reduce skin-flap survival. The purpose of this study was to determine if the preoperative administration of pentoxifylline can counteract the deleterious effects of nicotine on skin-flap survival in the rat. Sixty rats were distributed into four groups (n = 15). The survival of modified McFarlane skin flaps was assessed on postoperative day 7. The administration of nicotine (0.6 mg/kg) for 24 weeks preoperatively produced an average skin-flap survival of 59 percent; this was significantly decreased compared with controls (p < 0.05). When similarly treated animals were given pentoxifylline (20 mg/kg) for 30 days preoperatively, the mean skin-flap survival improved significantly to 80 percent (p < 0.05). Withholding nicotine for 2 weeks preoperatively also was found to significantly improve skin flap survival to 73 percent (p < 0.05). Blood filterability was measured as an indicator of viscosity. The blood filterability in rats that received nicotine for 24 weeks was significantly decreased compared with controls (p < 0.05). Both the addition of pentoxifylline preoperatively and the withholding of nicotine for 2 weeks preoperatively were found to significantly improve blood filterability compared with rats that received nicotine alone for 24 weeks postoperatively (p < 0.05). PMID- 9207661 TI - Comparison of secondary ischemic tolerance between pedicled and free island buttock skin flaps in the pig. AB - We compared the secondary ischemic tolerance of 8 x 12 cm surgically denervated pedicled island skin flaps and skin free flaps raised contralaterally on the buttocks of 50 pigs. The pedicled flaps and free flaps were subjected to 2 hours of primary warm global ischemia followed by 12 hours of reperfusion and 0, 2, 4, 6, or 10 hours of secondary warm global ischemia (n = 10 flaps). Skin necrosis was assessed 7 days after secondary ischemia. Pedicled skin flaps tolerated up to 10 hours of secondary ischemia without skin necrosis. However, incidences of skin necrosis (partial and total) in free flaps subjected to 0, 2, 4, 6, or 10 hours of secondary ischemia were 0, 10, 50, 80, and 100 percent, respectively. In a separate experiment, skin blood flow and hematology were studied in contralateral pedicled flaps and free flaps (n = 20) subjected to 4 hours of secondary ischemia. The skin blood flow measured by 15-micron microspheres at 1.5 hours of reperfusion was significantly higher (p < 0.01, n = 20) in pedicled skin flaps than in skin free flaps (1.91 +/- 0.35 versus 0.67 +/- 0.53 ml/min/100 gm). Under an operating microscope, microthrombi were observed near the arterial and/or venous anastomoses in 8 of 20 skin free flaps but none in the pedicled skin flaps. We obtained venous blood samples by cannulation of the major venae comitantes in 12 of the 20 skin free flaps in which there was no thrombosis in the vascular pedicle for hematologic studies. The venous plasma level of thromboxane B2 was significantly higher (p < 0.05) in the skin free flaps than in their contralateral pedicled skin flaps (195 +/- 49 versus 124 +/- 30 pg/ml). In addition, venous hematocrit, hemoglobin concentration, and white blood cell count also were significantly (p < 0.05) higher in skin free flaps compared with their contralateral pedicled skin flaps. Taken together, these observations were interpreted to indicate that buttock skin free flaps in the pig were less tolerant of secondary ischemia compared with their contralateral pedicled skin flaps subjected to the same ischemic protocol, and this reduced ischemic tolerance in skin free flaps was associated with compromised skin blood flow, hemoconcentration, and thrombosis in the vascular pedicle. PMID- 9207662 TI - Myxoid liposarcoma of the scalp: case report and literature review. AB - Myxoid liposarcoma of the head and neck is an extremely rare entity. The scalp region represents a risk factor to the patient because the diagnosis is usually made late, and the surgeon must have a high index of suspicion for this entity because suctioning the tumor without taking a biopsy further delays an accurate diagnosis. PMID- 9207663 TI - Use of hydroxyapatite for reconstruction after surgical removal of intraosseous hemangioma in the zygomatic bone. AB - Two rare cases of intraosseous hemangiomas in the zygomatic bone that were repaired after surgical removal of the tumor with hydroxyapatite implants are reported. Case 1 is a 42-year-old woman, and case 2 is a 46-year-old man. They complained of swelling in the right cheek. An intraosseous tumor in the right zygomatic bone was observed in both patients. The tumor was resected, and hydroxyapatite was employed to repair the defect caused by removal. After 4 years since the operation in case 1 and 8 months in case 2, the patients show satisfactory cosmetic appearance. Hydroxyapatite implant was useful for reconstruction of the defect after surgical removal of hemangiomas in the zygomatic bone. PMID- 9207664 TI - A case of popliteal pterygium treated along with nerve expansion. AB - The presence of a short sciatic nerve in the free edge of a popliteal pterygium makes this syndrome a surgical challenge. We present a case of popliteal pterygium that was treated by nerve expansion. The range of motion of the patient's knee joint was between 30 and 120 degrees. A 75-cc tissue expander was placed under the sciatic nerve and filled with 5 cc of saline solution weekly. When a total of 60 cc was reached, wound dehiscence was observed, and the procedure had to be stopped. The maximum extension obtained was 160 degrees. Since the expansion process had to be stopped early, the elongation attained by expansion was less than expected. We conclude that the nerve expansion method can be used as a good alternative treatment modality for patients with popliteal pterygium. PMID- 9207665 TI - Repair of a long-standing coccygeal hernia and open wound. AB - Coccygectomy is a rarely used treatment for coccygodynia because of its high failure rate and various complications, such as bowel herniation, as seen in our patient. The limited number of literature articles available on coccygectomy discussed the few successes but failed to mention the complications and treatment of such. As a result of coccygectomy, our patient suffered from bowel herniation, unsuccessfully treated twice with prosthetic mesh. Bilateral gluteus maximus muscle flaps plicated in the midline reduced our patient's hernia successfully. This procedure appears to be an effective treatment for this postcoccygectomy complication. PMID- 9207666 TI - Intraoperative expansion of the palate by the tumescent technique. AB - Intraoperative rapid expansion of soft tissue of the palate is reported. Tissue is recruited by repeated injections of an isotonic solution in a tumescent-like technique, thus producing increasing swelling of the palatal soft tissue. Straight direct closure of palatal clefts and fistulas is achieved with minimal scarring at the midline, no secondary healing of raw surfaces, and satisfactory and encouraging results. PMID- 9207667 TI - Island mucochrondrocutaneous flap for reconstruction of total loss of the lower eyelid. AB - The methods usually employed for reconstruction of total lower eyelid loss include (1) tarsoconjunctival flaps from the upper eyelid and skin-graft cover and (2) chondromucosal grafts and local skin-flap cover. We report a technique for coverage of these defects with an island flap involving the full thickness of the ipsilateral nasal wall based on the dorsal (external) nasal vessels, terminal branches of the ophthalmic vessels. The advantages of this method are (1) the main components of the eyelid (skin, tarsus, conjunctiva) are reconstructed in a single short operation, even under local anesthesia, (2) there is one donor area that can be closed primarily without significant deformity, (3) the upper lid remains intact, and ectropion is improbable, and (4) the procedure involves a short hospitalization. As disadvantages we might mention bulkiness of the new eyelid and difficult dissection of the (subcutaneous) pedicle. PMID- 9207669 TI - Unilateral osteotomies for external bony deviation of the nose. AB - Most crooked noses, at the end of a regular rhinoplasty, seem well positioned in the midline. However, several months later, the nasal pyramid may begin its lateral migration and the original deviation, to the chagrin of the surgeon, is partially reproduced. An approach to the externally deviated bony pyramid is reemphasized, involving the performance of one or more osteotomies on only one side of the nose.During the course of a regular rhinoplasty, the bilateral lateral osteotomies displace the lateral nasal walls medially. However, if only a unilateral osteotomy is performed, the nasal dorsum will seem to deviate toward the opposite side. This paper discusses in further detail how to use unilateral osteotomies in the rhinoplasty of an externally deviated nose. Although the unilateral osteotomy technique is not a new concept, it did not receive so far the interest it deserves among surgeons. The unilateral osteotomy technique is an adjunctive solution to a complex problem. PMID- 9207668 TI - Custom surgical stent for naris stenosis. AB - A technique has been described that allows accurate fabrication of a surgical nasal stent, made preoperatively, based on a mirror image of the shape of the normal naris. The advantage to the surgeon is a surgical stent that has the greatest accuracy possible preoperatively, plus the ability to remake the stent postoperatively with even more accuracy should it be needed. This stent can then be worn long term with minimal visibility and maximum patient acceptance during the critical months postoperatively when relapse is most likely. PMID- 9207670 TI - The use of iodate stain to enhance contrast between vermilion and skin in lip reconstruction. PMID- 9207672 TI - A retrospective of personal craniofaciodental research and clinical practice. AB - Recent reports have emphasized the need for further knowledge about the growth of bone(s). This is particularly true because an understanding of the normal and abnormal growth of bone(s) forms the basis of early recognition and appropriate treatment of many deformities. It is less well appreciated, however, that the findings in the abnormal may be employed to test and extend our knowledge of the normal. Genetic makeup, as well as various types of diseases and injuries such as trauma, inflammation, radiation, and chemicals, may affect skeletal growth sites and centers, thereby causing faulty growth of bone(s). The degree of the subsequent deformity will depend not only on the type, intensity, extent, and chronology of the noxious agent but also on the site and its particular susceptibility and growth activity. The problem of treatment of craniofaciodental deformities is a difficult one. Over the years, I conceived, designed, initiated, and carried out a series of experiments in regard to bone(s), teeth, and cartilage in both young and adult animals (turtles, rats, gophers, lagomorphs, pigs, dogs, and monkeys). Eventually, I directed my efforts principally toward local surgical experimentation as it related to both normal and abnormal gross postnatal craniofaciodental growth. Because of the wide variety of different structures, their interrelated individualities, and the challenges presented in both its richness of sites of growth and complexity, the skull proved to be a most unusual source of study. The purpose of this selective, organized, and limited review, analysis, and summary of personally conducted experiments is to relate certain aspects of differential growth and change and nonchange to age, sites, rates, factors, and mechanisms. In many instances, correlations are made between research findings and clinical practice. No such similar report as this was found in the literature. This retrospective study brings it all together. PMID- 9207671 TI - The "round block" purse-string suture: a simple method to close skin defects with minimal scarring. AB - We report the use of a subcuticular purse-string suture for closure of surgical skin defects, a simple maneuver that we have found to be very useful in closing difficult wounds and reducing scarring. The purse-string suture is performed with a large nonabsorbable suture that is passed intradermally and left in situ at least 4 weeks. This technique has been applied in 196 patients for a total of 221 sutures over a period of 2 years, being used to close skin defects from 2 x 2 to 8 x 11 cm in many areas of the body. All the patients showed, at closure, a large number of concentric redundant folds as well as considerable distortion of nearby structures; both improved impressively over a period of 2 to 3 weeks and became nearly normal at the time of suture removal (4 to 8.2 weeks, mean 5.7 weeks). The initially very limited and almost circular scar oriented itself along the skin tension lines over a period of a few weeks and, when matured, was always shorter than the original defect. In general, minimal scar widening occurred when we used larger sutures (more than 0-1) that were left longer (more than 6 weeks). Complications have been 23 cases of dehiscence (10.4 percent) in 23 patients (between the fifth and sixteenth days, mean 6.7 days); they were caused by the bad quality of the skin and by the use of too small sutures that cut through the dermis. The "round block" suture has many advantages: 1. It is a simple, inexpensive, and rapid technique for closing wounds by expanding the surrounding skin and often avoiding the use of skin grafts and/or local flaps. 2. It can minimize scarring; the final scars are shorter than the original defect and usually of very good quality. 3. It allows a very useful temporary closure that stretches the surrounding skin while waiting for the definitive histologic report. If this method is not chosen as a definitive closure, later repair with local flaps may be facilitated. 4. For the reasons expressed above, it never compromises the final result even in cases of dehiscence. The main disadvantage is the acceptability of the method on the part of patients, who need to be carefully prepared for both the gross initial distortion and the long time the suture has to be retained; nevertheless, patient satisfaction with the final results in generally very high, especially in large excisions of the face. PMID- 9207673 TI - Refinements in endoscopic forehead rejuvenation. AB - Endoscopic forehead technique provides an effective method for rejuvenation of the upper face. Distinct advantages of this technique over classic methods of forehead rejuvenation such as coronal or subcutaneous approaches include significant reduction of incisional scars. Described here are three refinements related to (1) control of hair, (2) differential release of the periosteum, and (3) advanced fixation methods. Control of hair can be achieved simply by braiding and the use of an Endoscopic Access Device. Extensive release of the periosteum and arcus marginalis is recommended laterally, while elevating the medial periosteum either intact or with conservative release. Different and technologically more advanced fixation methods are described to provide better control of elevated forehead. Incorporation of these refinements strives to optimize aesthetic results while minimizing operative morbidity. These refinements have been implemented during the care of 29 patients and have proven to be of major value in achieving greater patient satisfaction and technical advancement. PMID- 9207674 TI - Internal stabilization of autogenous rib cartilage grafts in rhinoplasty: a barrier to cartilage warping. AB - Autogenous rib cartilage grafts have gained more widespread use in rhinoplasty as dorsal onlay grafts and columellar struts. However, the usefulness of rib as a donor site has been limited by difficulties with postoperative cartilage warping. We hypothesized that the internal stabilization of rib cartilage grafts with Kirschner wires would prevent warping. The costochondral cartilages of a fresh cadaver were harvested and carved into 4 x 10 x 40 nm blocks. A single 0.035-in K wire was placed longitudinally into the center of each of the study specimens (n = 9), whereas no internal stabilization was utilized in the control group (n = 9). Over a 10-day study period, a mean of 2.2 degrees of warping was observed in the grafts with K-wires as compared to 8.9 degrees in the control group. This indicates that internal stabilization of rib grafts significantly reduces warping (p < 0.001). In a subsequent clinical study, 28 patients underwent placement of internally stabilized columellar struts (n = 19) and/or dorsal nasal grafts (n = 12) using autogenous rib cartilage. At a mean follow-up of 13.5 months (range 3 to 36 months), graft warping was not observed in any patient. Satisfactory aesthetic results were achieved in all but one patient, in whom mild displacement of a dorsal onlay graft occurred. Palatal extrusion of the K-wire occurred in 3 of the first 9 columellar struts. This prompted an alteration in technique with no subsequent extrusions. We conclude that the internal stabilization of autogenous rib cartilage grafts with K-wires effectively prevents graft warpage. PMID- 9207675 TI - Simplified anatomic method of double-eyelid operation: septodermal fixation technique. AB - Conventionally, it has been argued that the aponeurosis meets the orbital septum below the upper tarsal border in Orientals. However, we have shown that the aponeurosis fused with the orbital septum above the upper tarsal border in 502 patients, which contradicts the classic concept. In 457 patients there were distinct layers of fascia anterior to the orbital septum that originate from the septum and insert onto the pretarsal aponeurotic expansion. In Oriental eyelids, the preaponeurotic fat and orbital septum hang below the fusion line of the septum and aponeurosis. The hanging portion of the septum was dissected from the aponeurosis, plicated, and sutured to the dermis of the pretarsal flap. We performed the septodermal fixation technique in 512 patients over 3 years starting in March of 1992. Complications and changes of the folds were analyzed. The technique produced much less edema and discomfort and created more natural fold lines than any other technique. Patients were very lightly satisfied with the shape of the folds that we were able to design and adjust precisely with this method. PMID- 9207676 TI - Microbial growth inside saline-filled breast implants. AB - In vitro and in vivo experiments were conducted to determine whether intraluminal saline in breast implants can support the growth of common wound-infecting microorganisms over a prolonged period of time. The bacteria tested were Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Corynebacterium jeikeium, Enterobacter cloacae, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Three fungal species also were tested: Aspergillus fumigatus, Paecilomyces variotii, and Candida albicans. In the in vitro study, four organisms survived in flasks of sterile saline for the 2 weeks in which serial cultures were performed: K. pneumoniae, C. albicans, A. fumigatus, and P. variotii. In the in vivo study, 61 white rabbits (122 implants) received both an experimental implant inoculated with one of the test organisms and a control implant containing only sterile saline. They were sacrificed at 1-, 3-, or 6 month scheduled endpoints. None of the control implants containing sterile saline had positive cultures. In contrast, the intraluminal saline was culture positive for 7 of the 10 inoculated organisms after varying lengths of time: S. epidermidis, E. coli, E. cloacae, K. pneumoniae, P. aeruginosa, A. fumigatus, and P. variotii. Samples of capsular tissue also were cultured. Of the 122 capsular tissue specimens, 21 (17 percent) had positive cultures and surrounded both inoculated and sterile implants. In most instances, capsules that were culture positive contained an organism different from the one that had been inoculated in the group. In only 3 cases was the same organism cultured from both the periprosthetic tissue and the intraluminal saline, and these may represent instances of the inoculated organism migrating through the implants filler valves. The data show that several types of bacteria (particularly gram-negative species) and fungi can grow and reproduce in a restricted saline environment for extended periods of time. PMID- 9207677 TI - Distribution of organosilicon polymers in augmentation mammaplasties at autopsy. AB - Silicone-containing breast implants have been used since 1963 for cosmetic augmentation and breast reconstruction. Currently, there is intense debate regarding the extent and mechanism of migration of silicone from the area of implant. The current study compares tissue distribution of organosilicon polymers between women with and without silicone breast implants to determine the extent of silicone migration from breast implants. Samples were collected at autopsy from 15 individuals with bilateral breast implants with no known history of chest trauma and from 14 age- and sex-matched controls. Capsule, breast, axillary lymph nodes, abdominal fat, liver, lung, and spleen were collected for analysis of organosilicon polymers by atomic absorption spectrometry and for examination by light microscopy. Blood was collected for analysis of rheumatoid factor and antinuclear antibodies. Silicone was observed microscopically in at least one capsule section from all implant cases and in at least one lymph node in 8 of 15 implant cases. Silicone was not observed in lymph nodes from control cases. Organosilicon polymers were extracted from tissue using heptane, and the silicon content of the extract was quantitated by atomic absorption spectrometry. Silicon was detected in all capsules; statistically significant increases of organosilicon polymers were measured in axillary lymph nodes, breast, and abdominal fat from individuals with silicone breast implants when compared with the nonimplant group. Measurable amounts of organosilicon polymers were found in tissues from the nonimplant group. Suitable blood specimens were analyzed for the presence of rheumatoid factor and antinuclear antibodies. All nine implant cases tested were negative for the presence of antinuclear antibodies. Three implant cases which were tested for rheumatoid factor also were negative. We conclude that organosilicon polymers routinely migrate from the site of breast implantation to regional tissues near the implant site. Tissues from nonimplant cases often contained measurable amounts of organosilicon polymers, and tissue distribution was variable within any single individual: this is consistent with the wide-spread use and form of organosilicon polymers. PMID- 9207678 TI - Aesthetic outcome of breast implant removal in 85 consecutive patients. AB - As we began to see increasing numbers of women concerned about their gel-filled breast implants, we became aware that we could not advise them with any degree of confidence what they might expect in terms of aesthetic result after implant removal. We decided to review the records and outcomes over a 2-year period of a number of patients who underwent implant removal. Eighty-five consecutive patients were reviewed, 69 of whom had undergone cosmetic augmentation and 16 of whom had breast reconstruction with silicone gel implant(s). Thirty-nine of the 69 cosmetic augmentation patients had removal of implants alone, and 27 had removal accompanied by mastopexy. Three had reaugmentation with saline-filled implants; one had replacement with saline-filled implants. Fifteen of the 16 reconstruction patients underwent autogenous tissue transfer. Preoperative and postoperative photographs of all patients were mixed randomly and rated by two independent raters in four aesthetic categories on a five-point scoring system. Repeatability was measured several weeks later, when each rater scored randomly selected photographs from this patient pool. The patients also performed their own outcome evaluations by means of questionnaire. We discovered that cosmetic augmentation patients who undergo implant removal only often suffer adverse aesthetic results. The postremoval appearance of many cosmetic augmentation patients actually will be improved over their preoperative appearance when mastopexy is performed in conjunction with implant removal. The study demonstrated that patients with certain body types could expect a particular outcome; i.e., women with asthenic builds and older patients with lax, striated breast skin generally had unsatisfactory aesthetic outcomes with implant removal only. Patients selected for autogenous breast reconstruction had favorable results, with extended latissimus dorsi and TRAM flaps yielding equally good outcomes. The study allows us to offer patients an optimistic view of postoperative results following breast implant removal. We have begun to advise selected patients that implant removal accompanied by mastopexy provides a more pleasing aesthetic outcome than implant removal alone. PMID- 9207679 TI - Ultrasonic liposculpturing: extrapolations from the analysis of in vivo sonicated adipose tissue. AB - The ultrasonic liposculpturing technique is currently gaining increasing popularity. Although ultrasound is an accepted part of our diagnostic medical practice, the way in which it interacts with solid living tissue is still a complex and unsolved biophysical problem. Very few studies, if any, have followed the effects of diffusion of this intriguing technique on the fields of biosafety and interaction mechanisms. We evaluate the results of our standard ultrasound liposculpturing technique in order to recognize the physical mechanism-thermal, cavitational, or "direct"-involved in the damaging process. Our microscopic analysis of sonicated adipose tissue confirms that ultrasound is highly selective in its action, producing disruption of macromolecules and cellular structures probably through microstreaming tissue movement. The results of ultrasonic liposculpturing and standard suction lipoplasty are compared. The main advantages of this new technique are the possibility of a very selective destruction of adipose tissue and the prospective solution to such delicate problems as the irregularity of cellulite. PMID- 9207680 TI - Standardization in photography for body contour surgery and suction-assisted lipectomy. AB - Despite the existence of basic clinical standards in plastic surgery, specific guidelines for body contour photography have not been detailed previously. In this report we propose standard and supplemental views for positioning of the subject for suction-assisted lipectomy and body contour surgery. Also demonstrated are specialty views for the face, where liposuction has become an integral component of the procedures. Finally, recommendations for photographic documentation of skin "textural" changes and "cellulite" improvement with liposuction, as well as regions requiring lipectomy, are discussed. A professional photographic studio and a model were utilized. Proper lighting, equipment, and backgrounds are described to achieve such standards. General principles for clinical photography are reviewed. We present standard and supplemental views for suction-assisted lipectomy and body contour surgery, with an emphasis on methods to address advanced liposuction techniques (i.e., superficial suction lipectomy) that may affect texture and contour of the skin. These techniques provide consistency for all practitioners, allowing comparison of results and techniques. PMID- 9207681 TI - Ultrasonically assisted lipectomy in aesthetic breast surgery. AB - A case report of a young patient with marked asymmetry treated successfully with ultrasonically assisted lipectomy with a good functional cosmetic result, undetectable scars, and mammographic control and showing no ill-effect on the breast parenchyma is presented. Further studies and follow-up are needed to confirm the value and advisability of using ultrasonic energy in the female breast. PMID- 9207682 TI - Arteriovenous fistula of the scalp secondary to punch autograft hair transplantation: angioarchitecture, histopathology, and endovascular and surgical therapy. AB - Arteriovenous fistula of the scalp secondary to punch autograft technique is a relatively uncommon occurrence, similar to traumatic scalp arteriovenous fistulas from other causes. A pulsatile subcutaneous mass with an associated thrill or bruit and symptoms including pain or headache is a common presentation. Angiography is required for full diagnostic evaluation. Angioarchitecture may appear complex, even with a single-hole fistula. Super-selective angiography and embolization facilitate surgery and provide essential information regarding angioarchitecture. Complete excision of the lesion is curative. Identification and resection of the draining vein is mandatory to ensure a complete resection. The lesion may extend across traditional anatomic planes. Ligation of proximal feeding arteries is inadequate and potentially harmful. Histopathology of the traumatic arteriovenous fistula may appear similar to that of an arteriovenous malformation. Acquired arteriovenous fistulas and congenital arteriovenous malformations are markedly similar in their ultimate histopathology, angioarchitecture, angiographic appearance, hemodynamics, and treatment requirements. They should be considered to represent a spectrum of the same disease state rather than discrete entities. PMID- 9207683 TI - External cantilever sling in septorhinoplasty: a new technique. AB - Development of mucoperichondrial and mucoperiosteal flaps bilaterally, total removal of the deviated septum, straightening it outside, followed by replacement as a free graft constitute an option in treatment of the severely deviated nose. Positioning and stabilization of the septum in this technique may prove insufficient in the majority of septorhinoplasty patients in whom both nasal bones are also immobilized. Positioning and stabilization can be achieved adequately by the use of the presented technique that involves passing a 3-0 nylon suture to suspend the cartilage replant to a plastic splint applied to the dorsum of the nose, one-third of which is taped over the realigned nasal bones. In the past 5 years, 45 patients have undergone septorhinoplasty using this technique. The minimum follow-up period was 6 months. No patient developed any major complications. Secondary correction was necessary to improve the aesthetic result in only one patient. The results in patients who underwent this surgical procedure suggest that the presented technique provides excellent results in severely deviated noses associated with major deviations of the septum but must be limited only to those whose deformity is so severe that other techniques will be insufficient to obtain the desired result. PMID- 9207685 TI - Intraoperatively controlled small-incision forehead and brow lift. AB - Placement of screws at the posterior aspect of the incision sites and the gradual placement and removal of staples behind the screws allow for a controlled and titrated elevation of the forehead and brows. This technique does require the patient's acceptance of the temporary placement of screws and staples into the scalp, which in my experience has not been a problem. By seating the patient upright on the operating table, the surgeon can intraoperatively make a direct evaluation and adjustments (removal or addition of staples). This approach leads to a more controlled elevation of the forehead and brows and the potential of a more symmetrical and satisfactory result. PMID- 9207686 TI - The time has come--redux. PMID- 9207684 TI - Sideburn reconstruction with an expanded supraauricular trapezoidal flap. AB - Although loss of the sideburn and temporal scalp with subsequent alopecia is relatively frequent as a consequence of traumatic, surgical, and iatrogenic processes, not many techniques of sideburn reconstruction have been reported. We present a two-stage technique to correct the long-sideburns when there is an associated temporal alopecic defect due to trauma or surgery by means of an expanded supra-auricular trapezoidal flap. The method can achieve not only a satisfactory appearance of the sideburn but also a normal capillary line from the occipital to the frontal region. PMID- 9207687 TI - Reapplying. PMID- 9207688 TI - An automatic pressure infuser you may already have. PMID- 9207689 TI - Use of a soft denture liner as a protective dressing for alveolar bone grafting. PMID- 9207690 TI - The retroangular flap for nasal reconstructions. PMID- 9207692 TI - Healing of ear defects: primary or secondary. PMID- 9207691 TI - Reinnervation of the cutaneous part of a free fibula flap. PMID- 9207693 TI - The nasolabial fold in facial rejuvenation. PMID- 9207694 TI - Heparin and antithrombotic efficacy. PMID- 9207695 TI - What Mary's mother told me. PMID- 9207696 TI - Whole blood loss in liposuction: a protocol for handling liposuction specimen. PMID- 9207697 TI - Failure of silicone gel breast implants: analysis of literature data for 1652 explanted prostheses. PMID- 9207698 TI - Superficial liposculpture of the face and neck. PMID- 9207699 TI - Central loop of the H reflex. Normal value and use in S1 radiculopathy. AB - The H reflex in the S1 spinal nerve has been used in electrodiagnosis of S1 radiculopathy for several years. Direct stimulation of the S1 spinal nerve has provided more complete information about the H reflex pathway by dividing it into its peripheral and central (or spinal) conduction portions. A previous study compared spinal nerve latency with the H reflex latency, demonstrating an abnormal S1 ratio in subjects with S1 radiculopathy, thereby suggesting slowing within the spinal segment of the nerve. No study, however, has established a normal value for the central (spinal) portion of the H reflex. We electrodiagnostically tested 20 subjects with normal clinical neurologic and musculoskeletal examinations to define a normal sample of the central loop of the H reflex in the S1 spinal nerve. The peak latencies of the M and H reflex responses were measured after a single stimulus to the S1 spinal nerve. The data obtained established 7 +/- 0.3 ms as the normal value for the interpotential latency difference (central loop) of the H reflex in the S1 spinal nerve in healthy subjects. Six patients with clinical and electromyographic evidence of S1 radiculopathy all had central loop latencies of > 8 ms. The normal value of the central loop of the H reflex suggested in this pilot investigation may, therefore, be used to allow for earlier and more accurate diagnosis of an acute S1 radiculopathy. PMID- 9207700 TI - H reflex amplitude. Effect of leg muscle activity. AB - The H reflex is a monosynaptic reflex whose latency can be used to evaluate pathology of the S1 nerve root and axon. Little has been written about the use of the H reflex amplitude in evaluating S1 nerve root and axon. Little has been written about the use of the H reflex amplitude in evaluating S1 nerve physiology. Although amplitude measurement of evoked potentials is useful in the evaluation of other sensory and compound muscle action potentials, this single case study shows it to be an unreliable parameter with which to evaluate the integrity of S1 nerve fibers, because muscle activity in antagonist and agonist affects the amplitude and in some instances abolishes it. PMID- 9207701 TI - Late facilitation of the human soleus H reflex induced by sustained isometric maneuver. AB - Studying the effect of spinal cord reinforcement maneuvers (SCRMs) on H reflex assists in understanding aspects of motor control. Our objective was as follows: (1) to elucidate the effects of four neck positions (neck resting at neutral position (control); passive hyperflexion of the neck; hyperextension of the neck with simultaneous abdominal contraction; and sustained active neck hyperflexion); (2) to evaluate the temporal changes of soleus H reflexes repeatedly evoked after a period of sustained neck flexion. We used a prospective, intrinsically controlled trial of the effects of these SCRMs on the H reflexes and M-responses in ten healthy volunteers. Pre- and postmaneuver measures included H reflex and M response latencies and amplitudes, H/M maximum amplitude ratio, and H threshold. The four maneuvers showed no significant effect on the H reflex or M-response measures. To investigate temporal changes in the H reflex amplitude, H reflexes were repeatedly evoked at two-minute intervals after a one-minute period of active neck flexion. The amplitude of the H reflex was enhanced (P = 0.0356; analysis of variance), and the post hoc least significant difference test was significant at four minutes postmaneuver. Peak magnitude of the H reflex occurred at four minutes after relaxation, and the response returned to pretest baseline at eight minutes. The results of this study document the time course of repeated H reflexes after SCRM, and the timing of the H reflex was found to be a contributing variable that should be considered in future study designs. PMID- 9207702 TI - Body cooling may not improve somatosensory pathway function in multiple sclerosis. AB - We tested the hypothesis that reducing core body temperature in subjects with multiple sclerosis (MS) improves the cortical somatosensory evoked potential (SEP) response. Twenty subjects with definite MS were compared with 20 subjects without neurologic symptoms or disease. SEPs were recorded with stimulation of the tibial and median nerves unilaterally at 3.1 and 6.1 Hz. The procedure was repeated after a cooking vest and hat reduced core body temperature by an average of 0.46 +/- 0.28 degrees C. No appreciable change in latency or amplitude of the SEP responses occurred in either the control or MS group with cooling. Although the amplitude of the cortical SEP response was less at the 6.1 Hz rate than at 3.1 Hz, there were no statistically significant differences between the MS and control groups or between stimulation rates with cooling. We conclude that, although some reports suggest symptomatic improvements during cooling in subjects with MS, this improvement may not be associated with changes in the SEP. PMID- 9207703 TI - Peroneal nerve motor conduction to the proximal muscles. An alternative approach to conventional methods. AB - By using a fixed 8-cm distance between the stimulating and recording electrodes, we describe a new approach to peroneal nerve motor conduction studies to the proximal muscles. The recording active electrodes were fastened over the tibialis anterior and peroneus longus, while the stimulating cathode was placed at the posterolateral border of the fibular neck. The skin temperature was maintained at or above 32 degrees C in the lateral surface of the proximal shin. In normal subjects, 81 deep and superficial peroneal nerves were tested to establish reference values. The latencies to the tibialis anterior and peroneus longus were 2.5 +/- 0.3 (range, 2-3) ms and 2.6 +/- 0.2 (range, 1.9-3) ms, respectively. The amplitudes of compound muscle action potential for deep and superficial peroneal nerves were 6.2 +/- 1.3 (range, 3.6-9.3) and 6.2 +/- 1.7 (range, 3.4-10.6) mV, respectively. Also, 17 adult onset diabetic patients were tested for comparison. We concluded that this alternative test to the conventional methods is accessible for electrophysiologic evaluation of peroneal neuropathy. PMID- 9207705 TI - Effective physical therapy for chronic obstructive pulmonary disease. Pilot study of exercise in hot spring water. AB - Respiratory function and arterial blood gas were examined before and after a two month exercise program performed in a pool filled with hot spring water in 22 patients (70.9 +/- 9.1 years of age) with stable chronic obstructive pulmonary disease (12 cases of bronchial asthma and 10 cases of pulmonary emphysema) treated at our hospital between 1991 and 1994. The ratio of forced expired volume in one second to forced vital capacity (FEV1%) was significantly increased after the exercise program (P < 0.05), whereas the ratio of forced vital capacity to predicted normal value (%FVC) did not change. In addition, a tendency toward an increase in peak flow without an increase in maximum expiratory flow at 25 and 50% (V25 and V50) was observed. Although PaO2 was not increased, PaCO2 was selectively decreased by the exercise program (P < 0.05). The changes in respiratory function and arterial blood gas were considered attributable to respiratory muscle training and small airway clearance. Exercise in a pool filled with hot spring water may be useful in treating chronic obstructive pulmonary disease. PMID- 9207704 TI - Autonomic dysfunction associated with locked-in syndrome in a child. AB - This is a report of a young boy with the unusual combination of autonomic dysfunction with locked-in syndrome following multiple shunt revisions for hydrocephalus. A review of the literature on autonomic dysfunction syndrome and the complex clinical picture of the combined syndromes in a pediatric patient are discussed. The marked effectiveness of treatment with carbidopa/levodopa over bromocriptine for both syndromes is noted. PMID- 9207706 TI - A simple walk test to guide exercise programming of the elderly. AB - Exercise training programs are usually based on a maximal exercise stress test; however, this test is often difficult and sometimes frightening to older persons. This preliminary study reports on a fixed-distance, submaximal walk test and compares its usefulness for exercise prescription to that of the traditional maximal stress test. Ten cardiac patients, with an average age of 72 years (4 men), had recently clinically indicated maximal graded stress tests. Within one week, each had the walk test, which consisted of walking three times up and back 100 feet in the hospital corridor (total of 600 feet) as rapidly as possible, with a blood pressure cuff on their arm and carrying the electrocardiogram cable. Resting and peak heart rate, blood pressure, symptoms, and exercise electrocardiograms were compared for the walk test v the maximal stress test. Oxygen consumption was calculated from the peak workload on the maximal stress test and from walking speed on the walk test. The peak heart rates after the walk test were within the target heart rate zone (70-85%) for exercise programming, as obtained from the maximal stress test, in all patients except one. The calculated peak oxygen consumption from the walk test was also within the training zone (60 80%) obtained from the maximal stress test in all patients except one. This pilot study shows that a submaximal, steady state timed walk of 600 feet can be a feasible method of providing the information for exercise programming, possibly avoiding the need for a maximal stress test. This walk test can be performed easily by health-related staff without sophisticated facilities in an inpatient rehabilitation unit or nursing home; however, further study with a larger number of patients is necessary before this method of exercise prescription can be recommended. PMID- 9207707 TI - Profiles of functional recovery in fifty traumatically brain-injured patients after acute rehabilitation. AB - Research to demonstrate the efficacy of head injury rehabilitation is important at a time when cost-containment efforts are intensifying. A useful tool that would predict the functional improvement during hospitalization and length of stay (LOS) of persons with traumatic brain injury would be of benefit to patients and their families, insurance carriers, and rehabilitation specialists. This study examines functional improvements made by 50 traumatic brain-injured patients admitted to the rehabilitation unit at the University of California, Davis, Medical Center (UCDMC) as measured by the UCDMC Davis Functional Status Measure (DFSM), which was adapted from the Functional Independence Measure (FIM). The DFSM incorporates additional items to provide a more thorough measure of skills to be rehabilitated. The purpose of this study was to compare scores and profiles on the DFSM items obtained by patients with LOS greater than and less than and equal to the median rehabilitation LOS (23 days). Relationships were explored among admission DFSM scores, LOS for rehabilitation, discharge destination, and functional outcome. Results indicate that patients admitted to the rehabilitation unit attained a similar profile or level of function by discharge, regardless of admission Glasgow Coma Scale scores or admission DFSM scores. There were no significant differences in admission Glasgow Coma Scale score, age, acute LOS, or discharge disposition between the LOS groups. There was a significant difference in median admission DFSM score in 26 of 31 categories between the LOS groups. There was a significant difference in median DFSM change (admission to discharge) in 24 of 31 categories between the LOS groups. The admission DFSM total score was inversely proportional to the length of stay, with a correlation coefficient of 0.78. DFSM change and admission to discharge was linearly correlated with LOS (R = 0.66). The DFSM documents functional outcome and measures gains during inpatient rehabilitation. The DFSM profile is helpful in predicting the LOS needed to achieve those gains. PMID- 9207708 TI - Orthotic management of gait in spastic diplegia. AB - Orthoses are the primary conservative treatment option for control of dynamic equinus in spastic cerebral palsy. Our purpose was to compare the effects of a fixed ankle-foot orthosis (AFO), a supramalleolar orthosis (SMO), and a no-brace condition, but including shoes. Gait analyses were performed on 11 children with spastic diplegia, using a system with four cameras and two concealed force plates. Ensemble averages of time-distance, kinematic, and kinetic parameters were obtained for each condition, and a repeated measures analysis of variance was performed (P < 0.05). Among the important findings were as follows: (1) AFOs significantly reduced ankle excursion, increased dorsiflexion angle at foot strike, increased plantar flexion moment in push-off, decreased ankle power absorbed during loading response, and decreased ankle power generated in push off; (2) SMOs did not restrict ankle range of motion or significantly alter the power and moment values at the ankle joint. Although neither brace changed stride length and walking speed, AFOs did offer some biomechanical benefits to the child with spastic diplegia, whereas SMOs appeared to have very little measurable effect. PMID- 9207709 TI - Functional outcome of quadruple amputees with end-stage renal disease. AB - A rare but catastrophic complication in end-stage renal disease (ESRD) patients with peripheral vascular disease is amputation of all four limbs secondary to gangrene. We present three patients with ESRD who underwent quadruple amputation. The purpose of this case study is to investigate the functional benefit of inpatient rehabilitation for such amputees. Our large, tertiary acute care hospital admitted 1,469 patients with ESRD during a continuous 63-month period. There were 72 amputation procedures: 57 involving lower limbs, and 15 involving upper limbs. Three patients had all four limbs amputated; these three were subsequently admitted to our acute inpatient rehabilitation center. Their median Functional Independence Measurement (FIM) score on admission was 52 and on discharge was 75. Their median length of stay was 24.5 days, which may be attributed to early postoperative therapy and de-emphasis on prosthetic replacement. At discharge, all three patients were able to perform sliding board transfers and self-propel wheelchairs modified with quad knobs and brake extenders. Each continued hemodialysis three times per week. Two patients were independent with feeding using adaptive equipment, and one required verbal cues. Two patients were able to write using dorsal wrist splints and pencil holders. One patient was able to use a speaker phone and lift lightweight objects with a left below-the-elbow hook-type prosthesis. Our review of these three cases demonstrates that inpatient rehabilitation can improve functional scores in quadruple amputee patients with ESRD. A large multicenter study is warranted to obtain adequate sample size to demonstrate statistical significance. PMID- 9207710 TI - Immunologic profile of patients with fibromyalgia. AB - Fibromyalgia is a musculoskeletal disorder characterized by generalized myalgias, arthralgias widespread tender points in discreet areas on examination. It is frequently accompanied by fatigue, stiffness, and a nonrestorative sleep pattern. These patients generally have a normal blood count and chemistry profile. There is a subset of people with fibromyalgia (FM) who test positive for the antinuclear antibody (ANA) and have constitutional symptoms that resemble those of patients with early lupus. We studied the immunologic profile of patients with FM who are ANA-positive (+). A retrospective review of patient records in a university-based rheumatology practice was conducted. In a group of 66 FM patients, 30% (20) were ANA+, with a 75% preponderance of the speckled pattern and 20% diffuse pattern. The remaining 5% were equally split between diffuse speckled and speckled-nucleolar patterns. All had negative staining for extractable nuclear antibodies. The Smart Index (SI), a ratio of the sedimentation rate to one-half the patient's age, was developed to characterize each patient's inflammatory response. The FM patients who were ANA negative (-) had a mean SI of 0.55, whereas the FM patient's who were ANA+ had a SI of 1.07. These ANA+ patients represent a subgroup of patients who have FM with an inflammatory response profile larger than that of the ANA-patients. PMID- 9207711 TI - Effect of age and activity on knee joint proprioception. AB - Falls lead to significant morbidity and mortality in persons older than 65 years of age. Impaired proprioception may be a contributing factor to falls, and this may be influenced by the level of habitual physical activity. The primary purpose of this study was to investigate knee joint proprioception among young volunteers and active and sedentary elderly volunteers. Knee joint proprioception was measured through reproduction of static knee angles using a Penny and Giles electrogoniometer. The secondary purpose of this investigation was to test the reproducibility of the Penny and Giles electrogoniometer in measuring static knee angles. Sixteen young subjects (age range, 19-27 years) and 24 elderly subjects (age range, 60-86 years) participated. Subjects were given a screening history and physical examination to exclude neuromuscular or vestibular disorders or lower limb injuries. Knee joint proprioception was measured two times during one week. The elderly group was separated into active and sedentary subgroups based on their level of activity during the past year. The electrogoniometer was placed laterally across the dominant knee joint. From the prone position each subject attained one of ten randomly predetermined knee joint angles from 10 degrees to 60 degrees. The subject then returned to the starting position and reproduced the test angle. The absolute angular error (the absolute difference between the test angle and subject perceived angle of knee flexion) was determined. A positive correlation was found between control visits for all subjects (r = 0.88), and significant differences were observed between young (mean, 2.01 +/- 0.46 degrees) and active-fold (mean, 3.12 +/- 1.12 degrees; P < 0.001), young and sedentary-old (mean, 4.58 +/- 1.93 degrees; P < 0.001), and active-old and sedentary-old (P < 0.03). These findings demonstrate that the Penny and Giles electrogoniometer is a reproducible device for measuring knee joint angles in both young and elderly subjects. Furthermore, we found that proprioception is diminished with age and that regular activity may attenuate this decline. One strategy to reduce the incidence of poor proprioception and fall with ageing may be regular exercise. PMID- 9207712 TI - Psychogenic polydipsia after traumatic brain injury. A case report. AB - Electrolyte abnormalities are common medical complications of traumatic brain injury (TBI). Hyponatremia is the most common of these disorders. The syndrome of inappropriate antidiuretic hormone and cerebral salt-wasting are the most well known causes of hyponatremia following TBI. In the presence of polydipsia and polyuria, psychogenic polydipsia should be included in the differential diagnosis. It is important to distinguish among these entities because treatment differs to such an extent that improper diagnosis may lead to a worsening of the patient's condition. We present a patient who presented with a new onset of polyuria and polydipsia after sustaining a TBI. Evaluation, including monitoring of fluid intake and output, serum and urine sodium and osmolarity, as well as a fluid deprivation test revealed the cause to be psychogenic polydipsia. The patient's symptoms improved after institution of a behavioral program and fluid restriction. Various models of drinking behavior have been used to identify the site of dysregulation. Dopaminergic, cholinergic, and hippocampal etiologies have been implicated in this abnormality of fluid homeostasis. If disorders of these systems can lead to psychogenic polydipsia, it is reasonable to believe that a person who has sustained a TBI would be at higher risk of developing psychogenic polydipsia. PMID- 9207713 TI - Rehabilitation after cardiac transplantation. Case series and literature review. AB - Twelve heart transplant recipients were admitted to the rehabilitation unit (RU) of a tertiary general hospital during a five-year period. Demographic, medical, and functional data were collected on these patients in a prospective and retrospective chart review. Functional status of each patient was assessed at both admission and discharge by means of the Modified Barthel Index (MBI). All transplant patients admitted to the RU were male, with an average age of 58 (range, 48-64) years. The mean MBI at admission was 57 (range, 31-75), and mean MBI at discharge was 86.5 (range, 55-100). The difference between the mean MBI admission score and that at discharge was demonstrated to be statistically significant (P < 0.001) using the paired t test. The average length of stay on the RU for the 12 patient cohort was 26 (range, 10-63) days. Ten of the 12 patients (83%) were discharged from the RU to the community. Two patients had to be transferred back to the acute care units after developing significant medical problems. Of the patients who returned to the community, the average number of medications at discharge was nine (range, 7-13), with all patients taking prednisone and cyclosporine. At the time of admission, all patients presented with numerous secondary medical problems. Six patients (50%) had hypertension, which required a medical regimen for control. Five patients had either inadequate oral intake or swallowing problems, thus requiring a feeding tube. In four of the five patients, the feeding tube was able to be removed during the RU course. Seven patients had associated neuromuscular deficits, which included hemiparesis (2 patients), paraparesis (1 patient), and myopathy (1 patient). Four patients were found to have pressure sores on admission to the RU, three of whom were completely healed by the time of discharge. Two of the patients had affective disorders that required follow-up by the psychiatry service during their stay on the RU. One patient was found to have radiographic evidence of a vertebral compression fracture but no other recipients had known fractures, osteoporosis, or osteopenia. As well as discussing the above data, the authors will also review basic exercise guidelines for cardiac transplantation patients. PMID- 9207715 TI - Inhibitors of angiotensin II: potential hazards for patients at risk for anaphylaxis? PMID- 9207714 TI - Roles of religiousness and spirituality in medical rehabilitation and the lives of persons with disabilities. A commentary. PMID- 9207716 TI - Epidemiology, pathogenesis, and treatment of the common cold. AB - OBJECTIVE: Reading this article will reinforce the reader's knowledge of the pathogenesis of the common cold. The rationale for current and potential therapies for the common cold are reviewed in the context of current concepts of the pathogenesis of these illnesses. DATA SOURCES AND STUDY SELECTION: A MEDLINE literature search was done using the search terms common cold, rhinovirus, and viral respiratory infection. The search was restricted to the English language. Articles were selected for review if the title and/or abstract suggested the content was relevant to the subject of this review. The bibliographies of selected articles were used as a source of additional literature. RESULTS: Recent studies suggest that the host response to the virus is an important contributor to the pathogenesis of the common cold. Inflammatory mediators, especially the pro-inflammatory cytokines, appear to be an important component of this response and present an attractive target for new interventions for common cold therapies. Currently available treatments for the common cold have limited efficacy against specific symptoms. These therapies should be selected to treat the specific symptoms that are perceived to be the most bothersome by the patient. PMID- 9207717 TI - Health risk assessment of fungi in home environments. AB - LEARNING OBJECTIVES: Reading this article will enable the readers to recognize the public health importance of fungi in the home environment. In view of the recognized impact of fungi on human health, the large population being exposed to fungi, and the large population risk for developing allergic diseases, there is a need to establish guidelines for allowable exposure to fungi based on a health risk assessment. The aim of this study was to evaluate the status of the data on the relationship between exposure to fungi in the home environment and allergic health effects with respect to the development of such guidelines. DATA SOURCES: The past 10 years of peer-reviewed literature focused on the relationships between respiratory disease and exposure to fungi in indoor environments was examined, Indexing terms included mold, fungi, allergy, asthma, and indoor environment, among others. Each study was evaluated on the following criteria: aim and design of the study, methods for assessing exposure and health effects, and data analysis. STUDY SELECTION: Nine population based studies were identified that examined the relationship between allergy and the presence of fungi in the home environment. These studies included quantitative measures of fungal presence in either air or dust. RESULTS: One or more positive associations were found between fungal levels and health outcomes in seven of the nine cross-sectional studies identified. CONCLUSIONS: Despite these positive associations it remains impossible to set guidelines for fungi in home environments based on health risk assessment. This is in part because of the cross-sectional study designs, and inconsistency and inadequate validation of the measures used to evaluate exposure and health effects. Future research designed to generate data that can be used for the development of health risk assessment based guidelines for fungi in home environments should focus on susceptible populations, and use measures that accurately represent exposure and adverse health effects. PMID- 9207718 TI - Chronic productive cough and bronchiectasis in a 40-year-old woman. PMID- 9207719 TI - Human lung anaphylaxis results in rapid release of interleukin-4. AB - BACKGROUND: Interleukin-4 has been implicated as having numerous roles in the inflammatory responses characteristic of allergic asthma. Interleukin-4 has been shown to be involved in IgE synthesis, upregulation of BCAM-1 on endothelium, and promotion of inflammatory cell infiltration into the airways. OBJECTIVE: We therefore examined whether IL-4 was produced after an IgE-mediated response in human lung samples. RESULTS: Anti-IgE treatment of 12 human lungs resulted in the significant release of IL-4 within 30 minutes. CONCLUSIONS: Although the source of released IL-4 is unknown, the rapid release of IL-4 suggests that cells with performed stores, such as mast cells and eosinophils, are involved. Once released, IL-4 may play an important role in the pathogenesis of asthma. PMID- 9207720 TI - Allergic contact reactions due to phenylephrine hydrochloride in eyedrops. AB - BACKGROUND: Dermatitis of and around the eye is common. Allergic contact reactions from phenylephrine are rare despite extensive use by ophthalmologists. Previous reports do not indicate crossreactivity between phenylephrine and other sympathomimetic drugs in patch testing. METHODS: We report three cases of allergic contact reactions (dermato-conjunctivitis) after eyedrops. Skin prick tests, epicutaneous testing with the implicated drugs, additives, and a complete patch test battery, TRUE test (Upjohn-Pharmacia, Sweden), were performed in each patient. RESULTS: All skin prick tests were negative. The three patients showed positive patch tests to phenylephrine and one of them also to ephedrine. Tolerance of the other eyedrops without phenylephrine was verified by challenge. CONCLUSION: Phenylephrine was the responsible agent for the reactions in our patients as confirmed by clinical findings and positive patch tests. Our findings suggest the central structure as the sensitizing part of drug in the second patient. Patch testing is essential for diagnosis of allergic contact reactions of and around the eye. PMID- 9207721 TI - Efficacy and safety of oral immunotherapy with short ragweed extract. AB - BACKGROUND: Oral immunotherapy, if proven safe and effective, could be an alternative to subcutaneous immunotherapy. OBJECTIVE: This pilot study investigated the clinic and immunologic effects of ragweed immunotherapy using a new microencapsulated, pH-sensitive, oral delivery system. METHODS: A double blind, placebo-controlled trial was conducted in 23 patients with allergic rhinitis to short ragweed. Following a baseline nasal challenge with ragweed allergen, oral immunotherapy with encapsulated short ragweed extract or placebo was administered once daily, 6 days/week. Dosed began at 3 micrograms Amb a 1 per day and were increased by 3 micrograms every three days as tolerated, to a maximum daily maintenance dose of 24 micrograms. A nasal challenge was repeated 6 weeks, later, followed by the continuation of maintenance therapy through the natural ragweed season. Daily allergy symptoms and relief medication usage was recorded. A final nasal challenge was performed at the end of the natural season. Short ragweed-specific serum IgE, IgG, and IgG4 antibody levels were measured every 2 weeks during the study. RESULTS: Maximum tolerated doses ranged from 6 to 24 micrograms Amb a 1 per day (74% reached 24 micrograms). Adverse events were not serious or different between the active and placebo groups. The active group showed increased in short ragweed-specific serum IgG and IgG4 antibody levels. Symptom scores during the natural season were numerically but not statistically lower in the active treatment group. This group also experienced a greater reduction from baseline in nasal reactivity as assessed by nasal challenge. CONCLUSIONS: These pilot data suggest that the encapsulated, pH-sensitive oral immunotherapy delivery system was safe, induced a brisk serologic response, and attenuated the symptomatic response to both experimental and environmental ragweed exposure. PMID- 9207722 TI - Inhibitory effect of cetirizine on cytokine-enhanced in vitro eosinophil survival. AB - BACKGROUND: Cetirizine is an antihistamine that inhibits in vivo eosinophil influx into the inflamed airways following allergen challenge, and in vitro eosinophil chemotaxis and adhesion. Since eosinophils are proposed to have an important role in the pathophysiology of asthma and allergic disease, the effects of cetirizine on eosinophil function may be a mechanism of this agent's therapeutic regulation of the allergic reaction. OBJECTIVE: To determine the effect of cetirizine on in vitro eosinophil survival. METHODS: Using human eosinophils isolated from patients with allergic rhinitis, the cells were cultured in vitro for 48 to 72 hours with medium, cetirizine, or dexamethasone in the presence of IL-5, IL-3, or GM-CSF. Eosinophil survival was assessed by trypan blue exclusion. RESULTS: In the presence of IL-5, but not GM-CSF or IL-3 100 microM cetirizine significantly inhibited eosinophil survival at 48 and 72 hours; the magnitude of this inhibition was dependent on cytokine concentration. Although cetirizine significantly suppressed cytokine promotion of eosinophil survival, it was not as potent as dexamethasone. CONCLUSIONS: Although the in vitro concentration of cetirizine was required to be quite high, cetirizine may affect in vivo airway inflammation through its inhibition of IL-5-dependent eosinophil survival. PMID- 9207723 TI - Airway fluoroscopic diagnosis of vocal cord dysfunction syndrome. AB - BACKGROUND: Vocal cord dysfunction syndrome is often misdiagnosed as refractory asthma. Airway fluoroscopy has recently been proposed as an alternative to laryngoscopy in the initial evaluation of certain cases of suspected vocal cord dysfunction. OBJECTIVE: To evaluate the use of airway radiographs and fluoroscopy in a patient with suspected vocal cord dysfunction. METHODS: We used soft tissue technique airway radiographs and fluoroscopy to evaluate the glottic function during inspiration and expiration in a 9-year-old boy with refractory asthma and suspected vocal cord dysfunction. RESULTS: The study confirmed paradoxical vocal cord motion. CONCLUSIONS: Airway radiographs and fluoroscopy provide a rapid and noninvasive means of diagnosing vocal cord dysfunction. Patients should still have laryngoscopy performed at the earliest possible moment to rule out the possibility of other laryngeal abnormalities. PMID- 9207724 TI - Seasonal variation of skin reactivity and specific IgE antibody to house dust mite. AB - BACKGROUND: House dust mite is an important cause of bronchial asthma worldwide. While the allergen is present all year-round, a seasonal variation of house dust mite allergen levels has been found. There have been few reports, however, on seasonal variation of specific immune response to house dust mite. OBJECTIVE: We studied the changes in skin reactivity and specific IgE antibody to house dust mite associated with seasonal variation of house dust mite allergen levels in houses of mite-sensitive asthmatic patients. METHODS: In 14 mite-sensitive asthmatic patients, house dust mite allergen (Der f 1) contents in bedding were measured monthly. Skin reactivity on prick test and specific IgE antibody to house dust mite, Dermatophagoides farinae (D. farinae), were determined every 3 months from july to December. RESULTS: The concentration of Der f 1 in dust from bedding reached maximum levels in August and September, and significantly decreased in November and December compared with August and September (P < .05). Skin reactivity (a ratio of mean wheal diameter of allergen to histamine) to D. farinae decreased significantly in December compared with September (P < .05). Serum levels of total IgE and specific IgE antibody to D. farinae decreased significantly in December compared with September (P < .05). CONCLUSION: These findings suggest that seasonal changes in natural exposure to house dust mite allergen might lead to concurrent changes in skin reactivity and specific IgE antibody to house dust mite in mite-sensitive asthmatic patients. PMID- 9207725 TI - Bevel-down superior to bevel-up in intradermal skin testing. AB - BACKGROUND: Intradermal skin testing is one of the most widely used procedures in the diagnosis of hypersensitivity diseases in vivo. It is critical to perform the test accurately and expediently. Yet, there are few articles describing its detailed technique or proficiency available. OBJECTIVES: To identify the better method for intradermal testing between the bevel-up and bevel-down techniques. METHODS: Three inexperienced testers performed intradermal injections using both methods. Four sets of paired trails each consisting of ten injections were applied randomly, alternating between the same volunteer subject's contralateral arms. Duration to complete ten injections was measured. Numbers of injection sites that bled, that squirted into the air, and failed to form a bleb were counted. The overall comfort level was determined. RESULTS: The time to complete the injections by bevel-up and by bevel-down methods were 165.5 +/- 31.3 and 152.5 +/- 27.4 seconds, respectively (P < .015). The number of injection sites that bled was higher in the bevel-up method, particularily on trial one (P < .001). Completion rate of successful bleb formation in bevel-down was 27.3 as compared with 23.3 in bevel-up method (P = .013). The comfort level was higher with the bevel-down than the bevel-up method (P = .0001). CONCLUSION: The bevel down method of intradermal testing is superior to the bevel-up method. PMID- 9207727 TI - Immunotherapy for allergy to fire ant venom. PMID- 9207726 TI - Intranasal budesonide spray as an adjunct to oral antibiotic therapy for acute sinusitis in children. AB - BACKGROUND: The role of topical corticosteroids in the treatment of acute sinusitis has not been established in children. OBJECTIVE: An attempt was made to determine the impact of topical corticosteroids as an adjunct to antibiotic treatment in the management of childhood sinusitis. METHODS: In a double-blind, placebo-controlled study, 151 children with sinusitis were recruited from a general pediatric outpatient clinic and 89 completed a 3-week trial. Treatment consisted of amoxicillin-clavulanate potassium, 40 mg/kg/d tid, combined with bid nasal spray of either budesonide, 50 micrograms, to each nostril (n = 43) or placebo )n = 46_ for 3 weeks. Patients maintained daily symptom cards throughout the study and were examined by the same physician each week. RESULTS: Clinical symptoms and signs decreased significantly in both treatment groups in comparison to baseline (P < .01). We detected a significant improvement in the scores of the cough and nasal discharge at the end of second week in the budesonide group when compared with placebo (P < .05). Friedman nonparametric repeated measures ANOVA test revealed a significant decrease in the total weekly scores of cough during the second week of budesonide treatment (P < .001) in contrast to continuous decline during the second and third weeks in the placebo group (P < .001 and P < .05, respectively). While the nasal discharge score decreased significantly during the second week in the budesonide group (P < .01), no significant effect on the nasal discharge score was observed in the placebo group. CONCLUSION: These data suggest that topical corticosteroids may be a useful ancillary treatment to antibiotics in childhood sinusitis and effective in reducing the cough and nasal discharge earlier in the course of acute sinusitis. PMID- 9207728 TI - Hair conditioner causes angioedema. PMID- 9207729 TI - Origin and characteristics of enteroinvasive strains of Escherichia coli (EIEC) isolated in Germany. AB - Thirty-five E. coli strains belonging to O-serogroups with enteroinvasive types of Escherichia coli (EIEC) isolated in Germany between 1989 and 1995 were investigated for invasivity-associated DNA sequences. Only 11 strains were positive for ipaH and thus confirmed as EIEC. All 11 EIEC isolates originated from human infections which were imported to Germany from Eastern Europe. EIEC O124 were most frequent and originated from asymptomatic Romanians arriving at Rostock, Germany in 1992 and 1993. In January 1993, EIEC O124 were isolated from faeces of a laboratory technician with diarrhoea working at the enteric pathogen department of the Institute of Hygiene in Rostock. By comparing her E. coli O124 isolate with recently imported O124 strains for Xba I restriction fragment length polymorphisms (RFLP) the probable source of infection could be determined. Four major RFLP patterns were found in the group of O124 strains. O124 strains with identical RFLP patterns were found in the group of 0124 strains. 0124 strains with identical RFLP patterns were isolated from people who were in close contact to each other. PMID- 9207731 TI - Isolation and characterization of group B streptococci from human and bovine sources within and around Nairobi. AB - Group B streptococci (GBS) were isolated from bovine milk and from vaginas and throats of antenatal and postnatal women using TKT and rapid GBS media. Sixty three of 529 (12%) bovine bulk milk samples, 9 of 48 (19%) vaginal and 3 of 48 (6%) throat samples were positive. Both bovine and human beta haemolytic isolates were characterized biochemically and serologically. Pigment production was a characteristic of both human and bovine beta haemolytic isolates. The majority (88%) of human isolates fermented salicin and not lactose and most bovine isolates were either lactose positive/salicin positive (41%) or lactose positive/salicin negative (38%). Human and bovine isolates were 100% and 85% typable respectively. Serotype distribution was similar in the bovine and human populations with serotype la, lc and lll being most common in both. Fermentation of sugars showed major differences between bovine and human isolates but similarity in serotype distribution suggests some genetic relationship. PMID- 9207732 TI - Vibrio vulnificus infection reporting on death certificates: the invisible impact of an often fatal infection. AB - This study assessed accuracy of (a) recording Vibrio vulnificus infection on death certificates and (b) International Classification of Disease (ICD)-9 codes for V. vulnificus. Patients with microbiologically confirmed V. vulnificus infection were identified as part of co-ordinated surveillance in four USA Gulf Coast states between 1989 and 1993. Of 60 deaths, 51 death certificates were reviewed and V. vulnificus was recorded as the immediate cause of death on 11 (22%). There was no ICD-9 code for V. vulnificus infection, thus no patients had an ICD-9 code indicating V. vulnificus infection. Of 23 certificates where V. vulnificus was recorded on the death certificate, only 5 (22%) were coded for Gram-negative, septicaemia. This study highlights the importance of teaching physicians how to provide epidemiologically meaningful data on death certificates and the need for accurate ICD mortality codes. PMID- 9207730 TI - Epidemic cholera among refugees in Malawi, Africa: treatment and transmission. AB - Between 23 August and 15 December 1990 an epidemic of cholera affected Mozambican refugees in Malawi causing 1931 cases (attack rate = 2.4%); 86% of patients had arrived in Malawi < 3 months before illness onset. There were 68 deaths (case fatality rate = 3.5%); most deaths (63%) occurred within 24 h of hospital admission which may have indicated delayed presentation to health facilities and inadequate early rehydration. Mortality was higher in children < 4 years old and febrile deaths may have been associated with prolonged i.v. use. Significant risk factors for illness (P < 0.05) in two case-control studies included drinking river water (odds ratio [OR] = 3.0); placing hands into stored household drinking water (OR = 6.0); and among those without adequate firewood to reheat food, eating leftover cooked peas (OR = 8.0). Toxigenic V. cholerae O1, serotype Inaba, was isolated from patients and stored household water. The rapidity with which newly arrived refugees became infected precluded effective use of a cholera vaccine to prevent cases unless vaccination had occurred immediately upon camp arrival. Improved access to treatment and care of paediatric patients, and increased use of oral rehydration therapy, could decrease mortality. Preventing future cholera outbreaks in Africa will depend on interrupting both waterborne and foodborne transmission of this pathogen. PMID- 9207734 TI - Sexually transmitted diseases among foreigners in Italy. Migration Medicine Study Group. AB - A sentinel surveillance system for the control of sexually transmitted diseases (STD) among foreigners was developed in Italy in 1991. From January 1991 to June 1995, 4030 foreigners with a new STD episode were reported. More than one-third of them were North-Africans. The most frequent STDs were non-specific urethritis and genital warts among men, and non-specific vaginitis and latent syphilis among women. The overall HIV prevalence was 5%, with large differences in rates in people from different continents. Very high HIV-positivity rates were observed among homosexuals and homosexual IDUs from Central-South America, with 39.1% and 77.8% seropositive individuals respectively. These data stress the need for increased knowledge of both the spread of risk factors for STDs among immigrants. Particular attention should be paid to counselling procedures focused on the prevention of risk behaviours for acquiring STDs and HIV infection. PMID- 9207733 TI - Comparative study of Mycobacterium paratuberculosis strains isolated from Crohn's disease and Johne's disease using restriction fragment length polymorphism and arbitrarily primed polymerase chain reaction. AB - To obtain insights into the pathogenic mechanisms involving Mycobacterium paratuberculosis in Crohn's disease (CD) we questioned if the strains of M. paratuberculosis isolated from CD are distinguishable from those involved in Johne's disease (JD), a chronic granulomatous enteritis in cattle. Accordingly we compared human and animal strains at the DNA level, both by the analysis of restriction fragment length polymorphism (RFLP) in and around the insertion sequence IS 900 and by the arbitrarily primed chain reaction (AP-PCR). Results are in favour of a common clonal origin for the 4 strains isolated from CD and for 8 of the 11 strains isolated from cattle and sheep JD. PMID- 9207735 TI - The role of young children in a community-wide outbreak of hepatitis A. AB - An Hasidic Jewish community has experienced recurrent hepatitis A outbreaks since 1980. To assess risk factors for illness during a 1985-6 outbreak, the authors reviewed case records and randomly selected 93 households for an interview and serologic survey. In the outbreak, 117 cases of hepatitis A were identified, with the highest attack rate (4.2%) among 3-5 year olds. Among the survey households, the presence of 3-5 year olds was the only risk factor that increased a household's risk of hepatitis A (indeterminant relative risk, P = 0.02). Furthermore, case households from the outbreak were more likely to have 3-5 years olds than were control households from the survey (odds ratio = 16.4, P < 0.001). Children 3-5 years old were more likely to have hepatitis A and may have been the most frequent transmitters of hepatitis A in this community. Hepatitis A vaccination of 3-5 year olds can protect this age group and might prevent future outbreaks in the community. PMID- 9207736 TI - An outbreak of rubella in British troops in Bosnia. AB - An outbreak of rubella in April 1996 involved four male British soldiers deployed to Bosnia-Herzegovina. All were helicopter ground crew who were members of the same unit and who periodically travelled to and worked at forward air refuelling stations in Bosnia. There was a potential for spread of the infection to adjacent British units, to troops of other nations in the peacekeeping force, and also to the local civilian population. The British force included 620 female personnel, some of whom may have been non-immune to rubella. One pregnant British servicewoman was repatriated to UK for her own protection. There was a potential health risk, including the possibility of congenital rubella syndrome, in the non immune wives and partners of deployed male personnel, as a result of contact during the mid-tour home leave of the husbands or partners. The outbreak was monitored through a medical surveillance system known as ARRC 97, and was contained by prompt and rigorous control measures. This outbreak shows the importance of effective surveillance and of good microbiology laboratory support during military operations. The role of immunization against rubella during future military deployments is discussed. PMID- 9207737 TI - The relationship between beta-2-microglobulin, CD4 lymphocyte count, AIDS and death in HIV-positive individuals. AB - The relationship, in 539 individuals infected with the human immunodeficiency virus (HIV), between two prognostic markers, the CD4 count and beta-2 microglobulin (B2M), and the development of the acquired immunodeficiency syndrome (AIDS) and death was investigated. Cox proportional hazards models were used to determine the risk of AIDS or death. In a multivariate model which adjusted for demographic factors and treatment, the most recent measurements of B2M (relative hazard (RH) 1.37 per g/l higher) and CD4 count (RH 2.17 per log unit lower) were both significantly associated with the development of AIDS. Similarly, in a multivariate model which additionally adjusted for the development of AIDS as a time dependent covariate, there was a strong relationship with risk of death for the most recent measurements of B2M (RH 1.34 per g/l higher), and CD4 lymphocyte count (RH 1.91 per log-unit lower). A difference in the level of B2M could be used among patients with similar CD4 counts as an indicator of increased risk of progression to AIDS or death. Using the most recent values of these markers provides a better estimate of the risk of AIDS or death, compared to the more common method of analysis, where baseline values of the markers are used. PMID- 9207738 TI - The relationship between abnormalities detected in live lambs on farms and those detected at post mortem meat inspection. AB - A prospective longitudinal study of diseases of lambs born in December and January and housed through to slaughter was carried out on three flocks (A, B and C) between 1989 and 1991. In the first year of the investigation (1989-90) three cohorts of approximately 80 lambs were examined in detail on a regular (weekly or fortnightly) basis. This involved over 2000 examinations and at least one clinical abnormality was observed in each lamb. In the second year (1990-1) the farmers were asked to present sick lambs for treatment on the farm. Farmers from flocks A and B participated in this part of the study; a total of 97/1295 lambs that were slaughtered received at least one treatment. The carcases and visceral organs of lambs from each flock were observed after slaughter. There was no association between the abnormalities observed during routine examination of the cohort lambs (year 1) and those observed at post mortem meat inspection. However, in year 2, in lambs from flock A, there was a significant association between lambs treated for arthritis or pneumonia on the farm and the presence of arthritic or pleuritic lesions, respectively, post mortem. In both years of the study lambs which were older when slaughtered were significantly more likely to have pleuritic, pneumonic or arthritic lesions at meat inspection. It was concluded that routine examination of groups of lambs is an inefficient and possibly ineffective method to identify lambs with lesions at slaughter. However, lambs which have been treated for disease, and the older lambs in a flock, had an increased prevalence of lesions post mortem and hence more detailed inspection of these animals would increase the efficiency of meat inspection. PMID- 9207739 TI - Schistosoma mansoni: development and modulation of the granuloma after or multiple exposures in the baboon (Papio cynocephalus anubis). AB - The ability of the host to modulate the granulomatous response around ova trapped in tissues determines the severity of disease to schistosome infections. Multiple factors may affect this modulation such as age, prior sensitization, history of treatment, and exposure. The present study examines the effect of different patterns of exposure on the sequential development and modulation of granuloma in juvenile Kenyan baboons (Papio cynocephalus anubis) after receiving either a single infection (SI) of 1500 Schistosoma mansoni cercariae or multiple infections (MI) of 150 cercariae, once a week for 10 weeks. Prior to sacrifice at 17 weeks postinfection (p.i.), liver biopsies were obtained at Weeks 0, 6, 9, and 13. SI animals experienced more prolonged dysentery and greater weight loss compared to MI animals. Peak hepatic granuloma size (mean 355 +/- 65.5 microns diameter), the maximum percentage of eosinophils in the granuloma (61%), and severity of disease occurred at 6 weeks in SI animals. Peak granuloma size and pathology did not appear until Week 9 in the MI animals (mean 317.7 +/- 67.3 microns diameter). Granuloma size, tissue eosinophilia, and gross pathology diminished by Week 13 p.i. and were virtually absent in both groups by Week 17. The decrease in granuloma size, pathology, and clinical illness resolved more rapidly in the MI baboons. Singly infected baboons were more susceptible to infection (83 +/- 12% of cercariae developed into adult worms) compared to MI baboons (67 +/- 7%, P < 0.01). Eggs recovered from tissues at necropsy were primarily confined to the large intestine (85% of total egg recovered), followed by liver (10%) and small intestine (5%). Significantly more eggs were recovered from MI compared to SI animals, indicating a higher fecundity of female worms in the MI baboons. These date demonstrate that granulomatous responses develop more slowly and modulate more rapidly with repeated infection compared to a single heavy infection and suggest the type of exposure may affect the pathologic response to infection. PMID- 9207740 TI - Plasmodium yoelii: resistance to disease is linked to the mtv-7 locus in BALB/c mice. AB - We have identified congenic mouse strains that differ dramatically in resistance to infection with the murine malaria parasite Plasmodium yoelii 17X. After infection, BALB/c mice develop severe anemia and a high degree of parasitemia which sometimes results in death. The mtv-7 congenic strain BALB.D2.mlsa, however, develops only a mild degree of anemia and parasitemia. In this paper we describe the course of the disease and discuss the potential role of mtv-7 and linked loci in control of this infection. These mice differ in their response to anemia, which may contribute to their differential susceptibility to disease; however, no influence of mtv-7 reactive T cells was documented. PMID- 9207741 TI - Ivermectin-induced killing of microfilariae in vitro by neutrophils mediated by NO. AB - Rat neutrophil granulocytes isolated after intraperitoneal casein injection of the donors exhibit high cytotoxic efficacy in vitro against microfilariae of Litomosoides carinii in the presence of ivermectin. Optimum effects of 80-90% killing of microfilariae were obtained with 100 ng ivermectin per milliliter and a microfilariae: cell ratio of 1:100. Spleen cells killed approximately 30% of the microfilariae under these conditions. Cytotoxic effects were independent of any adherence of the cell to the larvae. In contrast to the effects of spleen cells, cytotoxicity of neutrophils completely abrogated when cells and targets were separated by a membrane impermeable for the cells, suggesting a very short living mediator in the latter case. Correspondingly, cytotoxic effects of neutrophils were completely inhibited by the addition of the arginine analogues NG-monomethyl-L-arginine and L-canavanine, indicating the involvement of reactive nitrogen intermediates. The nitric oxide scavenger hemoglobin also protected the microfilariae. Several compounds which are known to interfere with reactive oxygen intermediates were ineffective. An excess of ferrous ions in the medium in the presence of a reducing agent significantly reduced the cytotoxic efficacy of neutrophils. PMID- 9207742 TI - ICAM-1 and iNOS expression increased in the skin of mice after vaccination with gamma-irradiated cercariae of Schistosoma mansoni. AB - Host responses to migrating schistosomula of Schistosoma mansoni were compared in the skin of naive, multiply infected, or vaccinated (with gamma-irradiated cercariae) mice during the first 72 hr after cercarial penetration. Cellular response to the migrating parasite was minimal in the skin of naive mice for up to 72 hr after infection. In sharp contrast, the multiply infected or vaccinated animals exhibited a marked inflammatory response in the skin as early as 8 hr after cutaneous penetration of the challenge cercariae. This early inflammatory response in the skin of sensitized animals was characterized by a significant increase in the number of infiltrating cells, predominantly mononuclear cells and neutrophils. Increased exudation of serum proteins was also present in the skin of sensitized animals in areas of cercarial challenge. A time course of analyses revealed that mononuclear cell numbers increased significantly in the skin of vaccinated animals as early as 60 min after a challenge infection and continued to be present at a significantly higher level up to 72 hr after challenge. Peak neutrophil responses occurred in the skin at 24 hr (in multiply infected animals) and at 48 hr (in vaccinated animals) after a challenge infection. Along with the massive cellular infiltration there was an increased tissue expression of ICAM-1 and mRNA for iNOS in the skin of sensitized animals. Further analysis showed that in sensitized animals increased ICAM-1 expression was predominantly found on endothelial cells lining dermal capillaries, especially in areas around schistosomular migration and on cells that surrounded schistosomula in the dermis. In naive animals, however, a similar infection did not induce any ICAM-1 expression or iNOS production in the skin. Thus, an ICAM-1 mediated early accumulation of mononuclear cells in the skin and local production of nitric oxide may be important for the initial cutaneous inflammatory immune responses to migrating schistosomula of S. mansoni in vaccinated animals. On the contrary, in naive animals a potential parasite-induced suppression of ICAM-1 may play an important role in reducing cellular reaction in the skin and consequently help the parasite evade immune responses in the skin. PMID- 9207743 TI - Uptake and cellular localization of exogenous lipids by Giardia lamblia, a primitive eukaryote. AB - Giardia lamblia trophozoites are unable to carry out de novo lipid synthesis. It is therefore likely that lipids are acquired from the small intestine of the host, in which the trophozoites are exposed to free and conjugated fatty acids, various sterols, phospholipids, bile acids, and bile-lipid mixed micelles. Here we show that G. lamblia is capable of taking up exogenous phosphatidylcholine (PC), phosphatidylinositol (PI), sphingomyelin (SM), cholesterol, ceramide (Cer), and fatty acids. Results from epifluorescence and high-resolution confocal microscopy suggest that fluorescent analogs of SM and PC were accumulated in the plasma membranes, whereas palmitic acid and Cer were localized intracellularly. Interestingly, many of these analogs were also concentrated in perinuclear regions. Similar labeling patterns were observed when the fluorescent analogs were delivered to the parasite via liposomes. To test whether G. lamblia was capable of esterifying exogenous fatty acids into membrane or cellular phospholipids, trophozoites were pulse-labeled with 3H-labeled palmitic or myristic acids and the phospholipids analyzed by thin-layer chromatography. Results document that G. lamblia was able to incorporate exogenous fatty acids into various phospholipids, i.e., PI, PC, PE, and PG. Interestingly, a major portion of radiolabeled fatty acids was incorporated into PG, a phospholipid characteristic of prokaryotic membranes. PMID- 9207744 TI - Trypanosoma cruzi: resistance to the pore forming protein of cytotoxic lymphocytes--perforin. AB - The pore-forming protein perforin is one of the main effector molecules which cytotoxic lymphocytes utilize to kill their targets both in vivo and in vitro. Natural killer cells and cytotoxic T lymphocytes play an important role in host defense against a number of intracellular microorganisms such as virus and protozoan, but the exact way they help control infection is unknown. On the other hand, many microorganisms have evolved successful escape strategies to avoid immune-cell-mediated attack. It is thus necessary to investigate the direct interaction of infectious microorganisms with the lytic machinery of cytotoxic lymphocytes and other cells. In the present work we report the effect of perforin on both a protozoan, Trypanosoma cruzi, and the infected host cell. Epimastigote, amastigote, and trypomastigote forms of T. cruzi, as well as infected macrophages, were assayed for their susceptibility to perforin based on three different criteria. T. cruzi in all three differentiation stages were resistant to purified perforin at doses up to 100-fold larger than that sufficient to kill susceptible tumor cells. No morphological change was observed under electron microscopy. Survival rates and infectivities of the treated parasites in vitro were similar to those of control parasites. Moreover, the measurement of calcium influx using Fura-2 to assess membrane damage revealed that T. cruzi resist perforin attack by avoiding transmembrane pore formation. Resistance to perforin was not transferred to host cells since infected macrophages could be easily destroyed by perforin while intracellular amastigotes remained intact. PMID- 9207745 TI - Experimental transmission of nocturnally periodic Wuchereria bancrofti to Indian leaf monkey (Presbytis entellus). AB - Successful experimental transmission of the human lymphatic dwelling nocturnal periodic strain of Wuchereria bancrofti has been achieved from man to Indian leaf monkey (Presbytis entellus) through a susceptible strain of Aedes aegypti. The prepatent period varied between 195 and 240 days. Microfilaria (mf) levels were in general low and the peak count was attained at 1800 hr. Of the adult worms 5 12.5% were recovered from male langur while females revealed poor (2.54%) recovery. This host--parasite model thus makes available the target parasite material in substantial quantity to work on chemotherapeutic and immunological investigations. PMID- 9207746 TI - Plasmodium falciparum: a simple, rapid method for detecting parasite clones in microtiter plates. PMID- 9207747 TI - Development of a rapid, nonradioactive, oligonucleotide-based assay for the detection of Setaria digitata. AB - Setaria species are filarial parasites which inhabit the peritoneal cavity of cattle and other ungulates. The parasite is generally considered to be nonpathogenic in its natural hosts, but the transmission of the infective larvae through mosquito vectors to its abnormal hosts (goats, sheep, or horses) can result in a serious and often fatal neuropathological disorder commonly referred to as cerebrospinal nematodiasis. We have previously described the cloning and characterization of a repetitive DNA sequence of Setaria digitata that could be used as a diagnostic probe to detect the parasite in host and vector populations. Here we report the development of a rapid nonradioactive hybridization assay using an oligonucleotide probe based on the above cloned repetitive sequence. PMID- 9207748 TI - Low serum HDL-cholesterol is associated with raised tumor necrosis factor-alpha during ENL reactions. AB - The concentrations of serum lipids and tumor necrosis factor (TNF) were measured in leprosy patients across the spectrum of the disease and in erythema nodosum leprosum (ENL) patients at the onset of the reaction and after the reaction had clinically subsided. Lepromatous/borderline lepromatous (LL/BL) patients had significantly higher serum triglyceride and lower HDL-cholesterol levels; there was no such change in the tuberculoid/borderline tuberculoid (TT/BT) patients. The household contacts (HC) of the LL/BL patients also had significantly lower serum HDL levels. ENL patients during the acute phase of the reaction had significantly lower total, LDL-, HDL-cholesterol levels compared to the stable LL/BL patients, and these changes were reversible to pre-ENL levels after the reaction had subsided. Serum TNF levels were significantly higher in household contacts and in LL/BL patients but were not statistically different in TT/BT patients. Serum TNF levels were also significantly higher during the acute phase of ENL, and declined after the clinical remission of the reaction to levels comparable with those of LL/BL patients. There was a significant negative correlation between serum TNF and HDL-cholesterol levels during and after ENL reaction. However, there was no such correlation between TNF and total or LDL cholesterol levels in ENL patients. Our results suggest that the changes in HDL cholesterol metabolism are a specific part of the host response to lepromatous leprosy and to the ENL reaction and may be mediated by increased TNF production. PMID- 9207749 TI - Regional lymphadenitis following antileprosy vaccine BCG with killed Mycobacterium leprae. AB - Phase-II and extended Phase-II studies were conducted in three different sets of the population in Thiruthani Taluk, Chengalpattu District, South India, involving BCG and killed Mycobacterium leprae (KML) combination vaccines to ascertain the acceptability of the vaccines. In the Phase-II study, 997 healthy volunteers were vaccinated on individual randomization with one of the vaccines arms: BCG 0.1 mg + 6 x 10(8) KML, BCG 0.1 mg + 5 x 10(7) KML, BCG 0.1 mg + 5 x 10(6) KML, BCG, 0.1 mg or normal saline. Blood samples were taken and the serum was tested for antibody levels against phenolic glycolipid-I (PGL-I) and the 35-kDa protein of M. leprae. In this study, we observed regional suppurative adenitis in 6% (6 out of 100), 3% (3 out of 100), and 3% (3 out of 100) of the vaccinees in the BCG 0.1 mg + 6 x 10(8) KML, BCG 0.1 mg + 5 x 10(7) KML, and BCG 0.1 mg + 5 x 10(6) KML vaccine arms, respectively, in the 13-70 year age group. Earlier BCG scar status, skin-test reactions to lepromin-A, Rees' MLSA, and serum antibody levels against PGL-I and the 35-kDa protein did not help to identify the group at risk of developing suppurative adenitis. Suppurative adenitis appears to have a different relationship between the age of the subject and the dose of the vaccine. In order to overcome the problem of regional suppurative adenitis and to know the mechanism involved, an extended Phase-II study was conducted in similar groups of the population by reducing the BCG and KML doses, i.e., with BCG 0.05 mg + 6 x 10(8) KML, BCG 0.05 mg + 5 x 10(7) KML, and BCG 0.01 mg + 5 x 10(7) KML. Biopsy specimens were collected from lymph nodes of the suppurative adenitis cases and were subjected for culture and histopathological examination. The observations showed that regional suppurative adenitis could be reduced to 1% in the BCG 0.05 + 6 x 10(8) KML group, 0.5% in the BCG 0.05 + 5 x 10(7) KML group, and 0.5% in the BCG 0.01 + 5 x 10(7) KML group. This phenomenon of suppurative adenitis appears to be related to the total dose of mycobacterial antigens. Suppurative adenitis was seen by weeks 18 and 20 post-vaccination in the latter two lower doses; whereas it was seen by week 8 in the higher dose of the combination vaccines. No case of suppurative adenitis was observed in the BCG 0.1 mg group. Culture and histopathology ruled out the possibilities of progressive BCG infection and superadded infection. Considering the above results, BCG 0.05 mg + 6 x 10(8) KML was acceptable for a large-scale vaccine trial in South India. PMID- 9207750 TI - Effect of steroid therapy on parameters of peripheral autonomic dysfunction in leprosy patients with acute neuritis. AB - Recent electrophysiological studies on peripheral autonomic dysfunction in leprosy patients show a high prevalence of autonomic dysfunction as measured by abnormal vasomotor reflexes (VMR) and absent sympathetic skin response (SSR). Nothing is known about the reversibility of these autonomic parameters with treatment. Since there is evidence that small fiber function may be the most reversible component in neuropathies, we measured the effect of steroid treatment on autonomic parameters together with motor and sensory functions in leprosy patients with acute neuritis. Control subjects were investigated for repeatability testing of autonomic function. Due to a relatively high variability on repeat VMR testing in the controls, we defined a change in VMR testing as a change of > 30%. With this definition, the VMR of 14.8% of the patients improved, 75% remained unchanged, and 10.2% worsened. Absent SSR became positive in 16.6% and remained unchanged in 83.4%. Improvement in sensory motor functions was seen in 21.2% and 1.3% of the patients, respectively. PMID- 9207751 TI - A study on the effectiveness and safety of the WHO/MDT regimen in the northeast of Thailand; a prospective study, 1984-1996. AB - The aim of this prospective study was to determine the effectiveness and safety of the multidrug therapy as recommended by the World Health Organization (WHO/MDT) in 1982. One-hundred-eighty-eight newly diagnosed leprosy patients [130 paucibacillary (PB) and 58 multibacillary (MB) patients] from three provinces in northeastern Thailand were recruited into a study from April 1984 to March 1985. The study lasted until May 1996. The results showed that 182 patients finished their course of WHO/MDT, representing a treatment completion rate of 95%; 167 (122 PB and 45 MB) were released from surveillance (RFS); 82 PB patients were still available for follow up by the end of 1994 and 31 MB patients by May 1996. Two PB patients were diagnosed with a relapse, showing a relapse rate of 0.2 per 100 person-years at risk. After an average of 8 years of follow up, no MB relapses have been diagnosed. The proportion of patients with a WHO grade 2 disability among PB and MB patients increased from 4% and 8% at the start of treatment to 7% and 13% at last examination, respectively. It is concluded that the fixed-duration, 6-month WHO/MDT regimen for PB leprosy and the 24-month regimen for MB leprosy are effective, acceptable and safe, and that clinical activity, histopathological activity and/or a positive skin smear at release from treatment (RFT) have no bearing on the efficacy of the WHO/MDT regimens. The relapse rates are low and in accordance with most published data available to date. The importance of skin-smear services for a reliable classification (WHO PB/MB classification for control programs) is stressed. PMID- 9207752 TI - Daily multidrug therapy for leprosy; results of a fourteen-year experience. AB - Between 1980 and 1994, 67 new or relapsing leprosy patients were treated by daily administered multidrug regimens. Tuberculoid patients (23 TT/BT) received either bitherapy [rifampin + dapsone or clofazimine (RMP + DDS or CLO)] or tritherapy [RMP + DDS and/or CLO and/or ethionamide (ETH)] until clinical cure. Lepromatous patients (44 BB/BL/LL) received tritherapy (RMP + DDS and/or CLO and/or ETH) at least until bacteriological negativity. Of the 23 tuberculoid patients only one patient (5%) was cured at 6 months and about 70% needed between 6 and 24 months of treatment to obtain clinical cure (mean 19.5 months). In the 44 lepromatous patients, the achievement of bacteriological negativity was significantly linked to the initial bacterial index (BI), and it occurred after 2 to 7 years (mean 66.5 months) of multidrug therapy (MDT). The average BI decrease per year was 1.1+ during the first year, 0.9+ the second year, and then < 0.5+ per year. Reactional states significantly (p < 0.01) influenced the BI course: reversal reactions (RR) accelerated while erythema nodosum leprosum (ENL) delayed the BI decrease. Three of the 23 (13%) tuberculoid and 19 of the 44 (43%) lepromatous patients (p < 0.02) exhibited a RR and 18 of 44 (41%) lepromatous patients had ENL during MDT. A late RR (LRR) was observed in 1 (5%) and 6 (17%) of our tuberculoid and lepromatous patients, respectively, and 3 (8%) of our lepromatous patients suffered post-MDT ENL. No confirmed relapse has been observed within a follow-up period of 6 months to 7 years and 3 months [59 person-years at risk (PYR)] for TT/BT patients and of 4 months to 5 years and 10 months (100 PYR) for BB/BL/LL patients. When compared to the recommended WHO/MDT, it appears that daily MDT does not increase the clinical or the bacteriological cure rates either at 6 months in paucibacillary tuberculoid patients or at 2d years in multibacillary lepromatous patients. Moreover, as does the WHO/MDT, our regimens show a high frequency of reactional states both during and after treatment. This fact constitutes the main new problem of the actual treatment of leprosy. PMID- 9207753 TI - Differential development of CD4 and CD8 cytotoxic T cells (CTL) in PBMC across the leprosy spectrum; IL-6 with IFN-gamma or IL-2 generate CTL in multibacillary patients. AB - In the present study we evaluated the contribution of CD4 and CD8 T cells on the antigen-specific cytotoxic activity induced by whole Mycobacterium leprae in leprosy patients and normal controls (N) as well as the modulation of this activity by some cytokines. Peripheral blood mononuclear cells (PBMC) from N or from leprosy patients were stimulated with antigen in the presence or absence of cytokines for 7 days. M. leprae-stimulated PBMC were depleted of CD4 or CD8 antigen-bearing cells and employed as effector cells in a 4-hr [31Cr]-release assay against autologous M. leprae-pulsed macrophages. Our results demonstrate that both CD4 and CD8 T cells contribute to M. leprae-induced cytotoxic activity, with differences observed in paucibacillary (PB) and multibacillary (MB) patients. CD8-mediated cytotoxic activity is higher than that of CD4 cells in PB patients, while in MB patients CD4 cytotoxicity is predominant. Our data also demonstrate that the generation of CD4 and CD8 cytotoxic T lymphocytes (CTL) can be modulated differentially by interleukin-4 (IL-4), IL-6, gamma interferon (IFN gamma), or IL-2. Although MB patients developed the lowest CTL response, cytokines such as IL-6 plus IL-2 or IFN-gamma were able to generate both CD4 and CD8 cytotoxic T cells from MB patients. In PB patients, IL-6 plus IFN-gamma displayed the highest stimulation on CD8 effector cells. Thus, an important role may be assigned to IL-6, together with IL-2 or IFN-gamma, in the differentiation of M. leprae-specific CTL effector cells. PMID- 9207754 TI - Immunotherapy of lepromin-negative borderline leprosy patients with low-dose Convit vaccine as an adjunct to multidrug therapy; a six-year follow-up study in Calcutta. AB - The present report, which describes management of lepromin-negative borderline leprosy patients with low-dose Convit vaccine, is an extension of our earlier study on the treatment of lepromatous leprosy patients with low-dose Convit vaccine as an adjunct to multidrug therapy (MDT). The test Group I, consisting of 50 lepromin-negative, borderline leprosy patients, were given low-dose Convit vaccine plus MDT. The control group II consisted of 25 lepromin-negative, borderline leprosy patients given BCG vaccination plus MDT and 25 lepromin negative, borderline leprosy patients given killed Mycobacterium leprae (human) vaccine plus MDT. The control group III consisted of 50 lepromin-positive, borderline leprosy patients not given any immunostimulation but given only MDT. Depending upon the lepromin unresponsiveness, the patients were given one to four inoculations of the various antileprosy vaccines and were followed up every 3 months for 2 years for clinical, bacteriological and immunological outcome. All patients belonging to the test and control groups showed clinical cure and bacteriological negativity within 2 years. However, immunologic potentiation, assessed by lepromin testing and the leukocyte migration inhibition test (LMIT), was better in the test patients receiving low-dose Convit vaccine plus MDT than in the control patients receiving BCG vaccine plus MDT or killed M. leprae vaccine plus MDT or MDT alone. But the capacity of clearance bacteria (CCB) test from the lepromin granuloma showed poor bacterial clearance in the test patients. However, there was no relapse during 6 years of follow up. Two mid-borderline (BB) patients had severe reversal reactions with lagophthalmos and wrist drop during immunotherapy despite being given low-dose Convit vaccine. PMID- 9207755 TI - Dharmendra antigen but not integral M. leprae is an efficient inducer of immunostimulant cytokine production by human monocytes, and M. leprae lipids inhibit the cytokine production. AB - Killed integral Mycobacterium leprae, Mitsuda antigen, and chloroform-treated M. leprae, Dharmendra antigen (Dh-Ag), have been used for the classification of leprosy patients based on cell-mediated immunity. Heat-killed M. leprae also were used as a component of the Convit vaccine. Human blood monocytes were stimulated with M. leprae or Dh-Ag and their cytokine-inducing ability was compared. Monocytes were cultured in the presence of fresh human serum because of the efficiency of cytokine induction and the phagocytosis of M. leprae have been shown to be optimal in the presence of fresh serum. M. leprae and Dh-Ag were equally phagocytosed by monocytes. Dh-Ag was more potent than M. leprae in the induction of immunostimulatory/proinflammatory cytokines, interleukin-1 (IL-1), IL-6 and tumor necrosis factor (TNF). In contrast, a comparable level of IL-1ra, an immunosuppressive cytokine, was induced by M. leprae and Dh-Ag. The lipids extracted from M. leprae induced none of these cytokines by monocytes. Nevertheless, when monocytes were pretreated with the lipids followed by stimulation with Dh-Ag, productions of IL-1, IL-6 and TNF were all inhibited in a dose-dependent manner. However, the lipids did not inhibit the cytokine production induced by other stimuli including BCG and lipopolysaccharide. Moreover the lipids did not affect the production of IL-1ra. These results suggest that the lipids from M. leprae are responsible for the poor cytokine inducing ability of M. leprae, thus favoring their infection. These results also suggest that Dh-Ag rather than integral M. leprae may be useful as a vaccine candidate because Dh-Ag is able to induce a large amount of cytokines from monocytes. PMID- 9207756 TI - Effect of phorbol myristate acetate (PMA) and ionophore A23187 on interleukin-2 levels and proliferation of activated T lymphocytes from patients with lepromatous leprosy. AB - The immunodeficiency present in patients with lepromatous leprosy is characterized by the limited proliferation of T lymphocytes, and is explained in part by the impaired synthesis of interleukin-2 (IL-2). Diacylglycerol (DAG) and calcium produce the activation of PKC, ERK and JNK kinases, implying a normal IL 2 response. Phorbol esters, such as PMA, can substitute for DAG and are mitogenic to human T and B cells activating several cytokine-encoding genes. Ionophore A23187 increases calcium permeability across the cellular membrane to the cytosol of lymphoid cells and is considered a co-mitogen of T lymphocytes. Here we report that: 1) PHA-activated T lymphocytes from LL patients can be separated in vitro into two groups: a) responders (R) with a stimulation index (SI) of > 10 and (b) nonresponders (NR) with a SI of < 10. 2) The proliferative responses of cells from LL(R), LL(NR) and normal subjects were measured after being stimulated with: I, PHA, PMA, PMA + I PHA + PMA and PHA + PMA + ionophore (PPI). The most important result occurs in LL(NR) patients whose cells did not respond to PHA stimulation but increased to normal levels of proliferation when they were stimulated with PMA. Furthermore, the three groups, (NR, R and normals) strongly increased their responses when they were incubated with PPi. 3) Finally, Il-2 concentrations in the supernatants of cultures of T lymphocytes from LL(NR), LL(R) and controls were relatively low when they were incubated with PHA or PMA, but the addition of ionophore to PMA and the combination of PHA + PMA strongly increased the production of IL-2 in all of them, reaching the optimum IL-2 concentration when PPI is used. It can be concluded that the use of PMA, analogous to DAG, and ionophore A23187 (calcium increaser) in cultures of mitogen activated T lymphocytes from LL patients induced the expression of the IL-2 gene, thus correcting the inadequate proliferation of T cells from LL patients. PMID- 9207757 TI - Cytokine gene expression in the foot pad and spleen of BALB/cAJcl mice infected with M. leprae. AB - The cytokine mRNAs expressed in the foot pads and spleens of BALB/cAJcl mice infected with Mycobacterium leprae were studied by the reverse transcriptase polymerase chain reaction (RT-PCR) method using cytokine-specific primers for interleukin-1 alpha (IL-1 alpha), -2, -4, -6, -10, -12-(p40), gamma interferon (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha), and TNF-beta, and then for CD4 and CD8 markers. The pattern of cytokine gene expression in the foot pad which supports M. leprae growth was different from the expression in the spleen which does not permit M. leprae multiplication in mice. Before BALB/cAjcl mice were infected with M. leprae, IL-1 alpha and TNF-beta mRNAs were expressed physiologically in the foot pad while all of the cytokine genes examined were expressed in the spleen. In the foot pads of mice inoculated with M. leprae, in addition to the physiological appearance of IL-1 alpha and TNF-beta mRNAs, these signals were intensified. TNF-alpha expression was induced by the infection. On the other hand, in the spleens of mice inoculated with M. leprae, CD4 mRNA expression disappeared on day 1 of the infection, which was accompanied by the reduced expression of IL-2, -4, -6, and -12 mRNAs. The recovery of CD4 mRNA expression at a latter stage was accompanied by a corresponding increase of the cytokine mRNA expression. It was suspected that these results might permit restricted growth of M. leprae in the foot pads of normal mice. Furthermore, our study suggests that tissue-specific, local, immunologic characteristics are important in M. leprae growth. PMID- 9207758 TI - Pathologic changes in a tibial nerve with surviving M. leprae in a healed tuberculoid leprosy patient. AB - A tibial nerve from a disease-arrested borderline tuberculoid (BT) leprosy patient was dissected out and examined almost in its entirety using hematoxylin and eosin staining, a modified Fite's stain for acid-fast bacilli (AFB), solochrome cyanin stain for myelin, and van Gieson's stain for fibrous tissue. Fibrosis of the perineurium and epineurium and fibrous replacement of the nerve parenchyma, which was maximum at the ankle joint area, were seen. In focal areas inflammation was present, especially in the epineurium around blood and lymph vessels. Even 21 years after adequate antileprosy therapy, AFB were present in the endoneurium in all except 2 of the 10 segments of the nerve, evoking hardly an inflammatory reaction or other ill effects. It is pointed out that BT leprosy should also be considered a generalized disease, especially when there is peripheral nerve trunk involvement and, in such cases, a longer duration of currently available antileprosy therapy is advisable. Trauma to nerve trunk plays a major role in producing nerve destruction and paralysis. PMID- 9207759 TI - Leprosy in children. PMID- 9207760 TI - Influence of DDT exposure on susceptibility to human leprosy bacilli in mice. PMID- 9207761 TI - Regarding Ebenezer, et al.'s MB nerve histology in clinically diagnosed BT leprosy patients. PMID- 9207762 TI - "Umbilicated" lesions in histoid leprosy. PMID- 9207763 TI - Regarding Pinitsoontorn, et al.'s rapid village survey. PMID- 9207764 TI - A Streptococcus mutans Safari! PMID- 9207765 TI - The genetic contribution to dental maturation. AB - It has been established in the literature that there is a major genetic impact on tooth size (Potter et al., 1976; Corruccini and Sharma, 1985; Sharma et al., 1985), tooth morphology (Kraus and Furr, 1952; Biggerstaff, 1970), and root formation (Garn et al., 1960; Green and Aszkler, 1970). None of the studies concerning root formation, however, used the more advanced method of path analysis and model fitting to estimate genetic influence. The aim of the present study was to determine the genetic and environmental influence on dental maturation. Dental age scores were determined on panoramic radiographs of 58 pairs of twins--26 monozygotic (MZ) and 32 dizygotic (DZ)--with the method of Demirjian et al. (1973). No mirror-image effect was found between the sides of the same individual or between twin members, so dental maturation seems to be symmetrical for both left and right sides of the mandible. Correlation coefficients were significantly higher in MZ than in DZ twins, which suggests a genetic influence. Model fitting showed that the variation in dental age was best explained by additive genetic influences (A-component) (43%) and by environmental factors common to both twins (C-component) (50%). The specific environment (E component) added only 8% to the model. The importance of the common environmental factor can be explained by the fact that twins, being raised together, share the same prenatal, natal, and immediate post-natal conditions that are of importance for the formation of the teeth. PMID- 9207766 TI - Capsaicin-sensitive A delta fibers in cat tooth pulp. AB - How close a correlation there is between the conduction velocity and receptive properties of pulpal nerve fibers is still unclear. Our specific aims were to confirm whether: (1) capsaicin affects not only polymodal C fibers but A delta fibers as well, and (2) A alpha polymodal nociceptors exist in the tooth pulp. A total of 139 functional single cat tooth pulp nerve fibers was isolated for analysis, of which 21 were A beta, 37 C, and 81 A delta fibers. The A delta fibers were divided into two groups: One (n = 38) consisted of those fibers whose conduction velocities were more than 2.0 m/s both inside and outside the tooth pulp, and the other (n = 43) consisted of those fibers whose intrapulpal conduction velocities were less than 2.0 m/s, with extrapulpal conduction velocities greater than 2.0 m/s. We used 82 fibers to record the neural response following the topical application of capsaicin for 60 min at increasing concentrations (1 nM, 100 nM, 10 muM) through thin dentin. Six of 25 slow A delta, 10/20 C, and no A beta (0/11) or fast A delta (0/26) fibers responded to 1 nM or 100 nM of capsaicin. When the three concentrations of capsaicin solution were applied in turn, the electrical threshold and latency of A beta and fast A delta fibers did not change, whereas those of slow A delta and C fibers gradually increased. In 0/11 A beta, 0/26 fast A delta, 13/25 slow A delta, and 18/20 C fibers, the conduction was blocked reversibly or irreversibly following the application of 10 muM of capsaicin. The amplitude of the late component of antidromic action potential of fast A delta fibers decreased after the capsaicin application. No neural discharge could be recorded from 19 (3 A beta, 5 fast A delta, 6 slow A delta, and 5C) fibers following the application of a single high concentration of capsaicin (10 muM). A single low concentration of capsaicin (100 nM) activated only some slow-conducting fibers (0/4 A beta, 0/4 FA delta, 3/6 SA delta and 4/6 C). Response properties recorded from the remaining 18 fibers (3 A beta, 3 fast A delta, 6 slow A delta, and 6 C) were not changed following the application of the control vehicle. These results confirm that a low concentration of capsaicin has an excitatory effect on the response of slow pulpal A delta as well as C fibers, and that a high concentration of capsaicin blocks the conduction of slow A delta and C fibers as well as the terminals of fast A delta fibers in the pulp. PMID- 9207767 TI - Localization of nerve cells in the developing rat tooth. AB - Earlier studies have shown that mammalian tooth formation can take place in the absence of peripheral nerve fibers. This has been taken to indicate that neurons are not needed for mammalian tooth development. However, our recent localization of peripherin, which is a neuronal cell marker, has suggested that neuronal cell bodies may be associated with developing teeth. In this study, we have analyzed in vivo and in vitro the presence of neuronal cells in developing rat tooth germs. When E14 and E16 rat first molars (thickening of presumptive dental epithelium and bud-stage tooth germ, respectively) were cultured in vitro, peripheral trigeminal axons degenerated. However, with antibodies against peripherin and L1 neural cell adhesion protein, we detected neuronal cell bodies and their axons in the explants. Next, the expression of neurofilament light chain (NF-L) mRNAs was studied by in situ hybridization of embryonic E12 first branchial arches and tooth germs from initiation to completion of crown morphogenesis (E13, five-day post-natal teeth). NF-L transcripts were first seen at the bud stage (E15) next to the dental epithelium at the buccal side of the tooth germ. At the cap stage (E18), NF-L mRNAs were located under the oral epithelium at some distance from dental epithelium. These expression patterns correlate to the previous localization of peripherin-positive cells and suggest that NF-L expression also revealed neuronal cells. Taken together, these results demonstrate that, in addition to projections of peripheral neurons, neuronal cells are associated with the developing teeth. Hence, it is possible that neuronal cells may participate in the regulation of mammalian tooth formation. PMID- 9207768 TI - Expression of connexin 43 in rat mandibular bone and periodontal ligament (PDL) cells during experimental tooth movement. AB - Bone remodeling in response to force requires the coordinated action of osteoblasts, osteoclasts, osteocytes, and periodontal ligament cells. Coordination among these cells may be mediated, in part, by cell-to-cell communication via gap junctions. This study tests the hypothesis that the regulation of expression of connexin 43, a gap junction protein, is part of the transduction mechanism between force as applied to bone during orthodontic tooth movement and bone remodeling. To test this hypothesis, we examined connexin 433 expression in a rat model system of experimental tooth movement. To establish the model, we extracted maxillary first molars to initiate supra-eruption of opposing mandibular molars. The rats were killed at 0, 6, 12, 24, and 48 hrs post extraction. The mandibles were removed, demineralized, and embedded in paraffin. To localize connexin 43 protein and mRNA, we used a specific antibody for immunohistochemistry and a specific cDNA probe for in situ hybridization. Western and Northern blot analyses were used to assess the specificity of the connexin 43 antibody and cDNA probe, respectively. We found connexin 43 protein expressed by osteoclasts (++ ++) and periodontal ligament cells (++ +) in compression zones, and by osteoblasts (++ ++) and osteocytes (++ ++) in tension zones of the periodontal ligament. In addition, connexin 43 mRNA was found in some bone and periodontal ligament cells. Connexin 43 protein was found, by densitometric analysis, to be higher in the periodontal ligament after exposure to force compared with controls (P < 0.001). The number of osteocytes expressing connexin 43 48 hrs after molar extraction was also significantly greater in bone subjected to tension when compared with controls (P < 0.001). The results of this study support the hypothesis that connexin 43 plays a role in the coordination of events during experimentally induced alveolar bone remodeling. PMID- 9207769 TI - In vitro cellular aging stimulates interleukin-1 beta production in stretched human periodontal-ligament-derived cells. AB - Although the severity of periodontal disease is known to be affected by host age, the pathological role of aging in periodontal disease, and especially that attributable to trauma from occlusion, has not been well-characterized. Interleukin (IL)-1 beta is a key mediator involved in periodontal diseases, a potent stimulator of bone resorption. Furthermore, it is produced by human periodontal ligament (PDL) cells in response to mechanical stress. To investigate the age-related changes in the biosynthetic capacity of IL-1 beta in PDL cells, we examined the effects of in vitro cellular aging with mechanical stress on IL-1 beta protein and gene expression by human PDL cells. Human PDL cells (young = 5th or 6th passage; old = 18-20th passage) were cultured on flexible-bottomed culture plates, and the cells were deformed at 6 cycles per min at 2 steps of tension force for 1 to 5 days. We found a two-fold increase in IL-1 beta production by old PDL cells subjected to mechanical tension compared with that by young PDL cells, although the constitutive levels of IL-1 beta were similar in both the young and old PDL cells. This increase was tension-dependent. IL- 1 beta mRNA was also detected in both cell types under basal conditions, and its expression was further enhanced by application of mechanical tension by use of reverse transcription-polymerase chain-reaction (RT-PCR) and in situ hybridization methods. The increase in signal rate was higher in the old cells than in the young cells. IL-1 beta-converting enzyme mRNA remained unchanged. It is possible that a large amount of IL- 1 beta produced by PDL cells from an aged host in response to mechanical force may be positively related to the acceleration of alveolar bone resorption. PMID- 9207770 TI - Identification of Bacteroides forsythus in subgingival dental plaque with the aid of a rapid PCR method. AB - Bacteroides forysthus has been shown to be prevalent among patients with periodontitis. Conventional microbiological methods used to identify this bacterium, however, are laborious and time-consuming and are therefore not well suited for screening purposes. We have developed a polymerase chain-reaction (PCR) method which is rapid, specific, and simple to perform and does not require other sample pre-treatment except a brief centrifugation. This method was applied to the detection of B. forsythus in subgingival plaque of 58 periodontitis patients. When compared with the results of conventional culturing, the PCR method always confirmed the culture-positive results, while none of the PCR negative samples was shown to be culture-positive. The PCR method appeared to give more than double the number of samples positive for B. forsythus than culturing (89.7% vs. 37.9%). The analysis requires less than 4 hrs to perform, and is specific only to B. forsythus and sensitive enough to detect fewer than 5 bacteria. PMID- 9207771 TI - Effect of amine and stannous fluoride on human neutrophil functions in vitro. AB - Amine fluoride (AmF)- and stannous fluoride (SnF2)-containing products were found to have a therapeutic effect on gingivitis and periodontitis. This effect was suggested to correlate with the antibacterial activity of the fluoride compounds. However, their effect on inflammatory cell function can also play a role in the therapeutic effect on gingival inflammation. The present study was designed to test the effects of AmF, SnF2, and an AmF/SnF2 combination on the function of human peripheral blood neutrophils, as compared with effects of chlorhexidine and salicylic acid. Neutrophils were isolated from human blood by ficoll centrifugation followed by dextran sedimentation. The neutrophils were pre incubated with AmF, SnF2, or AmF/SnF2, followed by stimulation with fMLP. Cell vitality was verified by trypan-blue exclusion (> 95% vitality at all tested concentrations). Superoxide production was measured by cytochrome C reduction and the enzymatic activity of lysozyme and beta-glucoronidase by optical density measurement of substrate conversion. The results showed that AmF, SnF2, or AmF/SnF2 enhanced by two- to three-fold the superoxide release from fMLP stimulated human neutrophils. Furthermore, the effective concentration of the AmF/SnF2 combination was several-fold lower than that of AmF or SnF2 alone (10 nM for AmF, 0.5 microM for SnF2, and 3 pM for SnF2/AmF). On the other hand, chlorhexidine and salicylic acid were found to reduce superoxide production by the cells. All the tested compounds had no effect on granular enzyme release by the stimulated neutrophils. The results suggest that AmF and SnF2 enhance the oxygen-dependent antibacterial activity of neutrophils. This effect may contribute to a more efficient elimination of bacteria from the periodontal environment, resulting in improvement in gingival health. PMID- 9207772 TI - Longevity and cariostatic effects of everyday conventional glass-ionomer and amalgam restorations in primary teeth: three-year results. AB - The aim of this study was to compare the longevity and cariostatic effects of everyday conventional glass-ionomer and amalgam restorations in primary teeth. The materials consisted of 515 Ketac-Fil glass-ionomer restorations and 543 Dispersalloy amalgam restorations prepared in 666 children, from 3 to 13 years of age, by 14 dentists within the Danish Public Dental Health Service in the municipalities of Vaerlose and Hillerod. The restorations, of which 79% were of the Class II type, were in contact with 593 unrestored surfaces in adjacent primary and permanent teeth. After 3 years, 6% of the patients had dropped out of the study, and 33% of the teeth were exfoliated with the restoration in situ. A further 37% of the glass-ionomer and 18% of th amalgam restorations were recorded as failed (p < 0.001). The frequency of failures was highest for Class II glass ionomer restorations, which showed a 50% median survival time of only 34 1/2 months, because of many fractures, while the 75% survival time for Class II amalgam restorations just exceeded the actual 36 months (p < 0.001). Caries progression was most often recorded in surfaces adjacent to amalgam restorations, and 21% of these surfaces needed restorative treatment vs. 12% of the surfaces adjacent to glass-ionomer restorations (p < 0.001). The three-year results indicated that conventional glass ionomer is not an appropriate alternative to amalgam for all types of restorations in primary teeth. In particular, the short longevity of Class II glass-ionomer restorations could not be compensated for by the reduced caries progression and need for restorative therapy of adjacent surfaces. PMID- 9207773 TI - Exposure to mercury vapor and impact on health in the dental profession in Sweden. AB - Possible adverse effects of mercury exposure in dentistry have been discussed in several studies. The objective of the present study was to carry out detailed measurements of mercury exposure in the dental profession in Sweden, and to search for adverse health effects from such exposure. We examined 22 dentists and 22 dental nurses, working in teams, at six Swedish dental clinics. Measurements of air mercury, performed with personal, active air samplers, showed a median air Hg of 1.8 micrograms/m3 for the dentists, and 2.1 micrograms/m3 for the dental nurses. Spot measurements with a direct reading instrument displayed temporarily elevated air Hg, especially during the preparation and application of amalgam. The average concentration of mercury in whole blood (B-Hg) was 18 nmol/L, in plasma (P-Hg) 5.1 nmol/L, and in urine (U-Hg) 3.0 nmol/mmol creatinine. Possible effects on the central nervous system (CNS) were registered with three questionnaires: Q16, Eysenck Personality Inventory (EPI), and the Profile of Mood Scales (POMS). In the Q16, the number of symptoms was statistically significantly higher in the dentistry group compared with an age- and gender-matched control group (n = 44). The urinary excretion of albumin and urinary activity of the tubular enzyme N-acetyl-beta-glucose-aminidase (NAG) did not differ between the two groups. The results confirm that exposure to mercury in the dental profession in Sweden is low. The air Hg levels were mainly influenced by the method of amalgam preparation and inserting, and by the method of air evacuation during drilling and polishing. PMID- 9207774 TI - In vitro wear of composite with varied cure, filler level, and filler treatment. AB - For the clinical wear of composite filing materials to be reduced, compositional factors such as degree of cure, filler level, and silanation level should be optimized. An oral-wear-stimulating machine was used to explore the effects of these factors on abrasion and attrition wear as well as on opposing enamel wear. The composites were made from Sr glass (1-2 micron avg) and a 50/50 Bis GMA/TEGDMA resin. Series I (A-D, E) were light-cured (Triad II) for 9, 12, 25, and 40 sec/side to produce degree of cure (DC) as measured by FTIR of 56, 60, 61, and 63%, respectively. E received an additional heat cure (120 degrees C for 10 min) to reach a DC of 66%. Series II (D, F-I) were filled to 62, 53, 48, 37, and 28 vol%, respectively. In series III (D, J-M), the portion of fillers treated with a silane coupler (MPS) was 100, 80, 60, 40, and 20%, respectively. Samples were cycled 50,000 times against an enamel antagonist in a poppy seed/PMMA slurry in the oral wear simulator to produce abrasion (load = 20 N) and attrition (load = 70 N) simultaneously. Wear depth (micron: n = 5) was measured by profilometry. Results for each series were analysed by ANOVA/Turkey's (p < or = 0.05). The wear depths did reflect cure values, though only the abrasion difference for E < A was significant. Greater wear was correlated with lower filler levels (r2 = 0.88; p < 0.05), significantly increasing below 48 vol% (G). Wear increased linearly as the percent of silane-treated fillers was reduced (r2 = 0.99; p < 0.05). Abrasion and attrition did not differ significantly for any composite. Wear of the opposing enamel was largely unchanged by these factors. Compositional factors including degree of cure, filler level, and silanation directly affected the wear resistance of dental composites evaluated in an oral wear simulator. PMID- 9207775 TI - Reaction strains on the condylar neck during mastication and maximum muscle stimulation in different condylar positions: an experimental study in the miniature pig. AB - Most researchers agree that the primate temporomandibular joint (TMJ) is loaded compressively during function and that condylar position must play a role in mediating such loads. However, the precise nature of that role remains unclear. Using a pig model in this study, we attempted to analyze strain on the neck of the condyle during normal mastication and during simulated function in different condylar positions. Miniature three-element rosette strain gauges were bonded to the lateral surface of the condylar neck in 4 female miniature pigs (one per condyle). Measurements of strain were made during normal mastication and with the pigs under general anesthesia during maximum stimulation of the masseter and temporalis muscles in each of five condylar positions--centric occlusion, centric relation, anterior, relaxed and wide open--established through use of acrylic splints. Condylar position was evaluated by superimposition of lateral and dorsoventral cephalograms, with measurement of horizontal and vertical changes in location of implants placed on the zygomatic arch. As in primates, the TMJ was found to be load-bearing during mastication, with compressive strain oriented approximately perpendicular to the occlusal plane. In 3 pigs, strain was higher during balancing than during working function. During stimulation, the TMJ reaction strains were significantly lower with the condyles in the anterior position compared with the other positions, and the compressive strain was directed more anteriorly along the neck of the condyle in that position. PMID- 9207776 TI - The Internet strikes back. PMID- 9207777 TI - Breast cancer genes--what are the real risks? PMID- 9207778 TI - Alagille syndrome--a notch up for the Notch receptor. PMID- 9207779 TI - Raf revealed in life-or-death decisions. PMID- 9207780 TI - Looms to weave genomic nets. PMID- 9207781 TI - FISH with a twist. PMID- 9207782 TI - And finally, genes for human obesity. PMID- 9207783 TI - The extent of genetic variation in the CCR5 gene. PMID- 9207784 TI - Reversal of a mitochondrial DNA defect in human skeletal muscle. PMID- 9207785 TI - Deletion of the promoter region in the Atp7a gene of the mottled dappled mouse. PMID- 9207786 TI - A mouse model for mitochondrial myopathy and cardiomyopathy resulting from a deficiency in the heart/muscle isoform of the adenine nucleotide translocator. AB - In an attempt to create an animal model of tissue-specific mitochondrial disease, we generated 'knockout' mice deficient in the heart/muscle isoform of the adenine nucleotide translocator (Ant1). Histological and ultrastructural examination of skeletal muscle from Ant1 null mutants revealed ragged-red muscle fibers and a dramatic proliferation of mitochondria, while examination of the heart revealed cardiac hypertrophy with mitochondrial proliferation. Mitochondria isolated from mutant skeletal muscle exhibited a severe defect in coupled respiration. Ant1 mutant adults also had a resting serum lactate level fourfold higher than that of controls, indicative of metabolic acidosis. Significantly, mutant adults manifested severe exercise intolerance. Therefore, Ant1 mutant mice have the biochemical, histological, metabolic and physiological characteristics of mitochondrial myopathy and cardiomyopathy. PMID- 9207787 TI - Mutations in the human Jagged1 gene are responsible for Alagille syndrome. AB - Alagille syndrome (AGS) is an autosomal-dominant disorder characterized by intrahepatic cholestasis and abnormalities of heart, eye and vertebrae, as well as a characteristic facial appearance. Identification of rare AGS patients with cytogenetic deletions has allowed mapping of the gene of 20p12. We have generated a cloned contig of the critical region and used fluorescent in situ hybridization on cells from patients with submicroscopic deletions to narrow the candidate region to only 250 kb. Within this region we identified JAG1, the human homologue of rat Jagged1, which encodes a ligand for the Notch receptor. Cell-cell Jagged/Notch interactions are known to be critical for determination of cell fates in early development, making this an attractive candidate gene for a developmental disorder in humans. Determining the complete exon-intron structure of JAG1 allowed detailed mutational analysis of DNA samples from non-deletion AGS patients, revealing three frame-shift mutations, two splice donor mutations and one mutation abolishing RNA expression from the altered allele. We conclude that AGS is caused by haploinsufficiency of JAG1. PMID- 9207788 TI - Alagille syndrome is caused by mutations in human Jagged1, which encodes a ligand for Notch1. AB - Alagille syndrome is an autosomal dominant disorder characterized by abnormal development of liver, heart, skeleton, eye, face and, less frequently, kidney. Analyses of many patients with cytogenetic deletions or rearrangements have mapped the gene to chromosome 20p12, although deletions are found in a relatively small proportion of patients (< 7%). We have mapped the human Jagged1 gene (JAG1), encoding a ligand for the developmentally important Notch transmembrane receptor, to the Alagille syndrome critical region within 20p12. The Notch intercellular signalling pathway has been shown to mediate cell fate decisions during development in invertebrates and vertebrates. We demonstrate four distinct coding mutations in JAG1 from four Alagille syndrome families, providing evidence that it is the causal gene for Alagille syndrome. All four mutations lie within conserved regions of the gene and cause translational frameshifts, resulting in gross alterations of the protein product Patients with cytogenetically detectable deletions including JAG1 have Alagille syndrome, supporting the hypothesis that haploinsufficiency for this gene is one of the mechanisms causing the Alagille syndrome phenotype. PMID- 9207789 TI - Padlock probes reveal single-nucleotide differences, parent of origin and in situ distribution of centromeric sequences in human chromosomes 13 and 21. AB - Chromosome centromeres, composed of repeated DNA sequences, orchestrate the correct segregation of chromatids in cell division. We have examined the centromeres of human chromosomes 13 and 21 by studying the distribution, in situ, of two alpha satellite sequences that differ in a single nucleotide position. This was possible using padlock probes, oligo-nucleotides that can be ligated into circles upon target recognition. The segregation of individual 13 and 21 homologues in a family was followed by monitoring of the signals from two differentially labelled probes, specific for either sequence variant. A characteristic arrangement of the repeat motifs in three separate spots, oriented transverse to the length axis of the metaphase chromosomes and bilaterally symmetric, indicates that only parts of the detected regions are involved in the centromeric region, joining the sister chromatids before anaphase. PMID- 9207790 TI - A component of the transcriptional repressor MeCP1 shares a motif with DNA methyltransferase and HRX proteins. AB - Methylation of cytosines within the sequence CpG is essential for mouse development and has been linked to transcriptional suppression in vertebrate systems. Methyl-CpG binding proteins (MeCPs) 1 and 2 bind preferentially to methylated DNA and can inhibit transcription. The gene for MeCP2 has been cloned and a methyl-CpG binding domain (MBD) within it has been defined. A search of DNA sequence databases with the MBD sequence identified a human cDNA with potential to encode an MBD-like region. Sequencing of the complete cDNA revealed that the open reading frame also encodes two cysteine-rich domains that are found in animal DNA methyltransferases (DNMTs) and in the mammalian HRX protein (also known as MLL and All-1). HRX is related to Drosophila trithorax. The protein, known as Protein Containing MBD (PCM1), was expressed in bacteria and shown to bind specifically to methylated DNA. PCM1 also repressed transcription in vitro in a methylation-dependent manner. A polyclonal antibody raised against the protein was able to 'supershift' the native MeCP11 complex from HeLa cells, indicating that PCM1 is a component of mammalian MeCP1. PMID- 9207791 TI - Frequent translocation t(4;14)(p16.3;q32.3) in multiple myeloma is associated with increased expression and activating mutations of fibroblast growth factor receptor 3. AB - Dysregulation of oncogenes by translocation to the IgH locus (14q32) is a seminal event in the pathogenesis of B-cell tumours. In multiple myeloma (MM), translocations to the IgH locus have been reported at an incidence of 20-60%. For most translocations, the partner chromosome is unknown (14q+); for the others, a diverse array of chromosomal partners have been identified, with 11q13 (cyclin D1) the only chromosome that is frequently involved. Recently, we developed a Southern-blot assay that detects translocation breakpoint fragments in most MM tumours, including those with no translocation detected by conventional karyotyping. In a continuing analysis of translocation in 21 myeloma cell lines and primary tumours, we show that the novel, karyotypically silent translocation t(4;14)(p16.3;q32.3) is present in five lines and at least three of ten primary tumours. The chromosome-4 breakpoints are clustered in a 70-kb region centromeric to the fibroblast growth factor receptor 3 gene (FGFR3), the apparent dysregulated oncogene. Two lines and one primary tumour with this translocation selectively express an FGFR3 allele containing activating mutations identified previously in thanatophoric dwarfism. We propose that after the t(4;14) translocation, somatic mutation during tumour progression frequently generates in FGFR3 protein that is active in the absence of ligand. PMID- 9207792 TI - Correlation between severity and SMN protein level in spinal muscular atrophy. AB - Spinal muscular atrophy (SMA) is a common autosomal recessive neuromuscular disorder characterized by degeneration of motor neurons of the spinal cord. Three different forms of childhood SMA have been recognized on the basis of age at onset and clinical course: Werdnig-Hoffmann disease (type-1), the intermediate form (type-II) and Kugelberg-Welander disease (type-III). A gene termed 'survival of motor neuron' (SMN) has been recognized as the disease-causing gene in SMA. SMN encodes a protein located within a novel nuclear structure and interacts with RNA-binding proteins. To elucidate the molecular mechanism underlying the pathogenesis of the disease, we examined the expression of the SMN gene in both controls and SMA patients by western blot and immunohistochemical analyses using antibodies raised against the SMN protein. The present study shows a marked deficiency of the SMN protein in SMA. PMID- 9207793 TI - Persistent and therapeutic concentrations of human factor IX in mice after hepatic gene transfer of recombinant AAV vectors. AB - Haemophilia B, or factor IX deficiency, is a X-linked recessive disorder that occurs in about one in 25,000 males, and severely affected people are at risk for spontaneous bleeding into numerous organs. Bleeding can be life-threatening or lead to chronic disabilities with haemophilic arthropathy. The severity of the bleeding tendency varies among patients and is related to the concentration of functional plasma factor IX. Patients with 5-30% of the normal factor IX have mild haemophilia that may not be recognized until adulthood or after heavy trauma or surgery. Therapy for acute bleeding consists of the transfusion of clotting factor concentrates prepared from human blood and recombinant clotting factors that are currently in clinical trials. Both recombinant retroviral and adenoviral vectors have successfully transferred factor IX cDNA into the livers of dogs with haemophilia B. Recombinant retroviral-mediated gene transfer results in persistent yet subtherapeutic concentrations of factor IX and requires the stimulation of hepatocyte replication before vector administration. Recombinant adenoviral vectors can temporarily cure the coagulation defect in the canine haemophilia B model; however, an immune response directed against viral gene products made by the vector results in toxicity and limited gene expression. The use of recombinant adeno-associated virus (rAAV) vectors is promising because the vector contains no viral genes and can transduce non-dividing cells. The efficacy of in vivo transduction of non-dividing cells has been demonstrated in a wide variety of tissues. In this report, we describe the successful transduction of the liver in vivo using r-AAV vectors delivered as a single administration to mice and demonstrate that persistent, curative concentrations of functional human factor IX can be achieved using wild-type-free and adenovirus-free rAAV vectors. This demonstrates the potential of treating haemophilia B by gene therapy at the natural site of factor IX production. PMID- 9207794 TI - Toward a functional analysis of the yeast genome through exhaustive two-hybrid screens. AB - The genome of the yeast Saccharomyces cerevisiae is now completely sequenced. Despite successful genetic work in recent years, 60% of yeast genes have no assigned function and half of those encode putative proteins without any homology with known proteins. Genetic analyses, such as suppressor or synthetic lethal screens, have suggested many functional links between gene products, some of which have been confirmed by biochemical means. Altogether, these approaches have led to a fairly extensive knowledge of defined biochemical pathways. However, the integration of these pathways against the background of complexity in a living cell remains to be accomplished. The two-hybrid method applied to the yeast genome might allow the characterization to the network of interactions between yeast proteins, leading to a better understanding of cellular functions. Such an analysis has been performed for the bacteriophage T7 genome that encodes 55 proteins and for Drosophila cell cycle regulators. However, the currently available two-hybrid methodology is not suitable for a large-scale project without specific methodological improvements In particular, the exhaustivity and selectivity of the screens must first be greatly improved. We constructed a new yeast genomic library and developed a highly selective two-hybrid procedure adapted for exhaustive screens of the yeast genome. For each bait we selected a limited set of interacting preys that we classified in categories of distinct heuristic values. Taking into account this classification, new baits were chosen among preys and, in turn, used for second-round screens. Repeating this procedure several times led to the characterization of the network of interactions. Using known pre-mRNA splicing factors as initial baits, we were able to characterize new interactions between known splicing factors, identify new yeast splicing factors, including homologues of human SF1 and SAP49, and reveal novel potential functional links between cellular pathways. Using different cellular pathways as anchor points, this novel strategy allows us to envision the building of an interaction map of the yeast proteome. In addition, this two-hybrid strategy could be applied to other genomes and might help to resolve the human protein linkage map. PMID- 9207796 TI - Griscelli disease maps to chromosome 15q21 and is associated with mutations in the myosin-Va gene. AB - Griscelli disease (OMIM 214450) is a rare autosomal recessive disorder characterized by pigmentary dilution, variable cellular immunodeficiency and onset of acute phases of uncontrolled lymphocyte and macrophage activation, leading to death in the absence of bone-marrow transplantation. The pigmentary dilution is characterized by a diffuse skin pigmentation, silvery hair, large clumps of pigments in the hair shafts (Fig. 1) and an accumulation of melanosomes in melanocytes, with abnormal transfer of the melanin granules to the keratinocytes. Immunological abnormalities are characterized by absent delayed type cutaneous hypersensitivity and an impaired natural-killer cell function. A similar disorder has been described in the dilute lethal mouse--which, however, differs by the occurrence of a severe neurological disorder. The dilute locus encodes myosin-Va, a member of the unconventional myosin family. Myosins bind actin and produce mechanical force through ATP hydrolysis. Some members of this family are thought to participate in organelle-transport machinery. Because of the phenotype resulting in the dilute mouse and because of their potential role in intracellular transport, unconventional myosin-encoding genes were regarded as candidate genes for Griscelli disease. Here we report that the Griscelli disease locus co-localizes on chromosome 15q21 with the myosin-Va gene, MYO5a, and that mutations of this gene occur in two patients with the disease. Griscelli disease is therefore a human equivalent of dilute expression in the mouse. PMID- 9207795 TI - Fringe boundaries coincide with Notch-dependent patterning centres in mammals and alter Notch-dependent development in Drosophila. AB - In both vertebrate and invertebrate development, cells are often programmed to adopt fates distinct from their neighbors. Genetic analyses in Drosophila melanogaster have highlighted the importance of cell surface and secreted proteins in these cell fate decisions. Homologues of these proteins have been identified and shown to play similar roles in vertebrate development. Fringe, a novel signalling protein, has been shown to induce wing margin formation in Drosophila. Fringe shares significant sequence homology and predicted secondary structure similarity with bacterial glycosyltransferases. Thus fringe may control wing development by altering glycosylation of cell surface and/or secreted molecules. Recently, two fringe genes were isolated from Xenopus laevis. We report here the cloning and characterization of three murine fringe genes (lunatic fringe, manic fringe and radical fringe). We find in several tissues that fringe expression boundaries coincide with Notch-dependent patterning centres and with Notch-ligand expression boundaries. Ectopic expression of murine manic fringe or radical fringe in Drosophila results in phenotypes that resemble those seen in Notch mutants. PMID- 9207797 TI - Endothelial apoptosis in Braf-deficient mice. AB - Tyrosine kinase growth factor receptors and Ras/Raf/MEK/MAPK signalling have been implicated in the suppression as well as augmentation of programmed cell death. In addition, a Ras-independent role for Raf as a suppressor of programmed cell death has been suggested by the recent finding that Craf1 interacts with members of the Bcl-2 family at mitochondrial membranes. However, genetic studies of C. elegans and Drosophila, as well as the targeted mutagenesis of the murine Araf gene, have failed to support such a role. Here we show that mice with a targeted disruption in the Braf gene die of vascular defects during mid-gestation. Braf -/ embryos, unlike Araf -/- or Craf1 -/- embryos (L.W. et al., unpublished), show an increased number of endothelial precursor cells, dramatically enlarged blood vessels and apoptotic death of differentiated endothelial cells. These results establish Braf as a critical signalling factor in the formation of the vascular system and provide the first genetic evidence for an essential role of Raf gene in the regulation of programmed cell death. PMID- 9207798 TI - Partial rescue of Brca1 (5-6) early embryonic lethality by p53 or p21 null mutation. AB - Mutations in the mouse Brca1 gene cause lethality at different embryonic stages. We have shown that Brca1 mutant embryos, in which the fifth and sixth exons of Brca1 are deleted die before E7.5 and show decreased cellular proliferation. Brca1 mutants also show decreased expression of mdm2, a gene encoding an inhibitor of p53 activity. Thus, we have proposed that the reduction in mdm2 expression in Brca1 (5-6) mutants might lead to increased p53 activity. Consistent with this finding, the expression of p21, which encodes a G1 cell cycle inhibitor and is a target for p53 transcriptional activation was dramatically increased in the Brca1 (5-6) mutants, suggesting that impaired cellular proliferation could be due to a G1 cell-cycle arrest, caused by increased p21 levels. To test this hypothesis, we generated mice double mutant for Brca1 (5-6) and p53, or Brca1 (5-6) and p21. Mutation in either p53 or p21 prolonged the survival of Brca1 (5-6) mutant embryos from E7.5 to E9.5. The development of most Brca1 (5-6): p21 double-mutant embryos was comparable to that of their wild-type littermates, although no mutant survived past E10.5. The fact that mutation of neither p53 nor p21 completely rescued Brca1 (5-6) embryos suggests that their lethality is likely due to a multi-factorial process. PMID- 9207799 TI - Obesity and impaired prohormone processing associated with mutations in the human prohormone convertase 1 gene. AB - Human obesity has an inherited component, but in contrast to rodent obesity, precise genetic defects have yet to be defined. A mutation of carboxypeptidase E (CPE), an enzyme active in the processing and sorting of prohormones, causes obesity in the fat/fat mouse. We have previously described a women with extreme childhood obesity (Fig. 1), abnormal glucose homeostasis, hypogonadotrophic hypogonadism, hypocortisolism and elevated plasma proinsulin and pro opiomelanocortin (POMC) concentrations but a very low insulin level, suggestive of a defective prohormone processing by the endopeptidase, prohormone convertase 1 (PC1; ref. 4). We now report this proband to be a compound heterozygote for mutations in PC1. Gly-->Arg483 prevents processing of proPC1 and leads to its retention in the endoplasmic reticulum (ER). A-->C+4 of the intro-5 donor splice site causes skipping of exon 5 leading to loss of 26 residues, a frameshift and creation of a premature stop codon within the catalytic domain. PC1 acts proximally to CPE in the pathway of post-translational processing of prohormones and neuropeptides. In view of the similarity between the proband and the fat/fat mouse phenotype, we infer that molecular defects in prohormone conversion may represent a generic mechanism for obesity, common to humans and rodents. PMID- 9207800 TI - Missense mutations abolishing DNA binding of the osteoblast-specific transcription factor OSF2/CBFA1 in cleidocranial dysplasia. AB - Cleidocranial dysplasia (CCD) is an autosomal dominant disorder characterized by hypoplastic or absent clavicles, large fontanelles, dental anomalies and delayed skeletal development. The phenotype is suggestive of a generalized defect in ossification and is one of the most common skeletal dysplasias not associated with disproportionate stature. To date, no genetic determinants of ossification have been identified. CCD has been mapped to chromosome 6p21, where CBFA1, a gene encoding OSF2/CBFA1, a transcriptional activator of osteoblast differentiation, has been localized. Here, we describe two de novo missense mutations, Met175Arg and Ser191Asn, in the OSF2/CBFA1 gene in two patients with CCD. These two mutations result in substitution of highly conserved amino acids in the DNA binding domain. DNA-binding studies with the mutant polypeptides show that these amino acid substitutions abolish the DNA-binding ability of OSF2/CBFA1 to its known target sequence. Concurrent studies show that heterozygous nonsense mutations in OSF2/CBFA1 also result in CCD, while mice homozygous for the osf2/cbfa1 mull allele exhibit a more severe lethal phenotype. Thus, these results together suggest that CCD is produced by haploinsufficiency of OSF2/CBFA1 and provide direct genetic evidence that the phenotype is secondary to an alteration of osteoblast differentiation. PMID- 9207802 TI - Endocervical curettage, cone margins, and residual adenocarcinoma in situ of the cervix. AB - OBJECTIVE: To evaluate endocervical curettage (ECC) and cone margin involvement in the management of adenocarcinoma in situ of the cervix. METHODS: Forty-two women with adenocarcinoma in situ without any associated invasive component underwent 49 cervical conizations. The ECC, cone margin involvement, and residual endocervical glandular disease were evaluated in a retrospective descriptive study. RESULTS: The patients ranged from 18 to 65 years old, with a median of 34 years and a mean of 37 years. Nineteen of 42 (45%) of the women presented with initial cervicovaginal cytology suggesting endocervical glandular abnormality. Twenty-seven patients (64%) had mixed lesions of adenocarcinoma in situ and squamous dysplasia noted in their cervical biopsy, conization, or hysterectomy specimens. Forty ECCs were performed at colposcopy or immediately after conization; 28 patients with ECCs subsequently underwent conization, and 12 underwent hysterectomy. Residual adenocarcinoma in situ was found in 18 (67%) of the 27 patients with negative ECCs and in ten of 13 women with positive ECCs. Residual adenocarcinoma in situ was found in two of seven patients with negative cone margins and seven of ten patients with positive margins. CONCLUSION: We found that negative ECCs and uninvolved cone margins in patients with cervical adenocarcinoma in situ were not reassuring of the absence of residual endocervical glandular disease in subsequent surgical specimens. Conservative management and subsequent surveillance of adenocarcinoma in situ should be undertaken with caution. PMID- 9207801 TI - Mutations in human TBX3 alter limb, apocrine and genital development in ulnar mammary syndrome. AB - Ulnar-mammary syndrome is a rare pleiotropic disorder affecting limb, apocrine gland, tooth and genital development. We demonstrate that mutations in human TBX3, a member of the T-box gene family, cause ulnar-mammary syndrome in two families. Each mutation (a single nucleotide deletion and a splice-site mutation) is predicted to cause haploinsufficiency of TBX3, implying that critical levels of this transcription factor are required for morphogenesis of several organs. Limb abnormalities of ulnar-mammary syndrome involve posterior elements. Mutations in TBX5, a related and linked gene, cause anterior limb abnormalities in Holt-Oram syndrome. We suggest that during the evolution of TBX3 and TBX5 from a common ancestral gene, each has acquired specific yet complementary roles in patterning the mammalian upper limb. PMID- 9207803 TI - Human papillomavirus infection in postmenopausal women with and without hormone therapy. AB - OBJECTIVE: To determine whether postmenopausal hormone therapy is associated with high-risk human papillomavirus (HPV) infection. METHODS: The detection rate of HPV DNA was studied in cellular residue from liquid-based collection tubes taken from 180 postmenopausal hormone users attending a menopausal clinic and 126 postmenopausal nonusers. The samples were analyzed with a hybrid capture technique using a mix of high- to intermediate-risk viral RNA probes. In all patients, information on potential confounding factors for HPV infection, including sociodemographic, reproductive, and gynecologic characteristics, was obtained. RESULTS: The prevalence of HPV DNA in this cohort of postmenopausal women was 1% (three of 306); only two of the 180 current users and one of the 124 nonusers tested positive. Only one of the three women with HPV-positive tests had lesional tissue (i.e., vulvar condylomata acuminata). The remaining two HPV positive women had negative cytology, colposcopy, and biopsy. In all three cases, viral burden was low, about 10 pg per cellular sample. The very low HPV prevalence precluded the analysis of correlation with age, ethnicity, education, sexual history, smoking, history of abnormal Papanicolaou smear, therapy for HPV related lesions, and contraceptive use. CONCLUSION: Identification of high-risk HPV types in post-menopausal women is rare, as detected by hybrid capture in cellular residue from the liquid-based cytology-collection system. Postmenopausal hormone therapy does not appear to promote viral replication or the risk of carrying high-risk HPV DNA or related lesional tissue in the lower genital tract. PMID- 9207804 TI - Predictors of bone mineral loss in patients with ovarian cancer treated with anticancer agents. AB - OBJECTIVE: To identify factors predicting bone mineral loss during anticancer chemotherapy. METHODS: Fifteen women (mean age 38.2 +/- 7.8 years; range 30-46 years) with ovarian cancer who had been treated with cisplatin-adriamycin cyclophosphamide for six cycles every 4 weeks following surgical cytoreductin were studied. Bone mineral density (BMD) of the lumbar spine (L2-L4) was measured by dual-energy x-ray absorptiometry before and after chemotherapy. Fifteen age matched women whose ovaries had been removed surgically for other reasons. served as controls. None of the patients had received hormonal treatment. The two groups were compared for percentage change of BMD (BMD%) over the same period. In the chemotherapy group, total fat mass, body fat ratio, total lean mass, percent lean, and ration of trunk fat to leg fat were measured by dual-energy x-ray absorptiometry. Lean loss during chemotherapy was also calculated. These variables were compared before and at the end of chemotherapy. Possible correlations of baseline variables with BMD% were determined in univariate and stepwise regression analysis. RESULTS: Mean ( +/- standard deviation) BMD decreased to 87.4 +/- 2.1% after six cycles of chemotherapy and 97.6 +/- 0.4% after 6 months in controls, but the greatest decrease was observed in the chemotherapy group (P < .001). Although baseline lean mass, baseline BMD, body weight, and lean loss during chemotherapy were correlated with BMD% in univariate analysis, baseline lean mass was still significant in stepwise regression analysis. CONCLUSION: Baseline lean mass predicts bone mineral lose with anticancer chemotherapy. PMID- 9207805 TI - Prevalence and correlates of breast and cervical cancer screening among older women. AB - OBJECTIVE: To identify and assess differences in cancer screening patterns among women 55-64, 65-74, 75-84, and over 84 years of age. METHODS: Nationally representative data reported in the 1990 Health promotion and Disease Prevention Supplement to the National Health Interview Survey of 28,584,574 women were analyzed secondarily. The dependent variables were a knowledge of breast self examination, over having had a mammogram, and a Papanicolaou smear within the last 3 years. Independent variables were age and various sociodemographic, health status, and health-belief measures. RESULTS: More than half (58%) of the women had ever had a mammogram, and of these, 91% had had between one and five mammograms. Over a third (35%) of those who had not had a mammogram attributed the omission to a lack of a recommendation by a physician. Almost half (45%) had had a breast examination by a physician within the last year, and 84% knew how to examine their own breasts. Approximately 87% had a Papanicolaou smear with the last 3 years. Age, race, education, and living in a large city were significantly associated with all three screening measures, but prevalent health beliefs were significantly associated only with breast-cancer screening. CONCLUSION: Lack of mammogram screening in a substantial number of women, attributed to lack of physician recommendation, decreased screening in the older age groups, and the negative association of three screening tests with race and residence in a large city suggest that new interventions are needed by health care providers and the public health community to increase older women's use of effective cancer screening techniques. PMID- 9207806 TI - Effect of estrogen on the size of low-density lipoprotein particles in postmenopausal women. AB - OBJECTIVE: To investigate the effects of estrogen on the size o low-density lipoprotein (LDL) particles in postmenopausal women. METHODS: We treated 20 postmenopausal women with 0.625 mg of conjugated equine estrogen daily for 3 months and measured the plasma levels of total cholesterol, triglyceride, high density lipoprotein (HDL), and apolipoproteins A-I, A-II, and B before and after therapy. We also analyzed concentrations of LDL cholesterol and LDL apolipoprotein B. The diameter of LDL particles was determined by gradient gel electrophoresis. RESULTS: Estrogen caused significant decreases in the plasma levels of total cholesterol and apolipoprotein B and significant increases in the plasma levels of triglyceride, HDL cholesterol, and apolipoprotein A-I and A-II. Mean levels of LDL cholesterol and LDL apolipoprotein B were reduced significantly (by 16.31%, P < .001, and 16.91%, P < .001, respectively) after estrogen treatment. Estrogen also significantly reduced the size of LDL particles, from 25.74 +/- 0.66 (mean +/- standard deviation) to 24.95 +/- 0.78 nm (P < .001). The LDL particle diameter correlated negatively with the plasma level of triglyceride (pre-treatment: r = 0.87, P < .001; post-treatment: r = 0.88, P < .001). Estrogen significantly increased the prevalence of LDL subclass pattern B, from 30 to 65% (P < .03). CONCLUSION: Estrogen affects lipid metabolism favorably by reducing the plasma concentration of LDL particles. Estrogen-induced increase in the plasma level of triglyceride appears to reduce the size of LDL particles. PMID- 9207807 TI - Double uterus, blind hemivagina, and ipsilateral renal agenesis: 36 cases and long-term follow-up. AB - OBJECTIVE: To review the experience of the Milan University First Department of Obstetrics and Gynecology in patients with double, didelphic, of bicornuate uterus, blind hemivagina, and ipsilateral renal agenesis, and to consider the frequently unsatisfactory surgical approach. METHODS: Thirty-six women with double, didelphic, or bicornuate uterus, blind hemivagina, and ipsilateral renal agenesis were identified from clinical records for the period 1962 to 1992. We evaluated demographic data, disease, symptoms, correctness of therapeutic approach, and definitive treatment. RESULTS: Seventeen patients previously had undergone incomplete surgery in other hospitals and 19 were treated by us for the first time. Total hysterectomy was performed on two of the 36 women and hemihysterectomy and hemicolpectomy were performed on four. In the other 30, the vaginal septum was excised and marsupialization was done. The pregnancy rate in the 15 women wanting children was 87% and the live birth rate was 77%. Serial biopsy specimens were obtained from the lateral fornix after the excision of the septum in 13 of the non-hysterectomized patients over 1-9 years and revealed progressively more extensive areas of squamous metaplasia of mullerian epithelium. In some isolated cases, papillary hyperplasia, mild dysplasia, and vaginal adenosis were found. At the end of follow-up, 16 patients still did not want children. Follow-up was possible in 34 cases. CONCLUSION: Early accurate diagnosis after menarche followed by excision and marsupialization of the blind hemivagina offers complete relief of symptoms and preserves reproductive potential. Partial morphologic changes are evident but metabolic modifications comparable to those of the adjacent normal vagina have not yet been documented. PMID- 9207809 TI - Prevalence of hydronephrosis in patients undergoing surgery for pelvic organ prolapse. AB - OBJECTIVE: To determine the prevalence of hydronephrosis in patients undergoing surgery for pelvic organ prolapse and to determine whether hydronephrosis is associated with the type and severity of prolapse. METHODS: The charts of 375 consecutive patients undergoing surgery for pelvic organ prolapse at the Cleveland Clinic Foundation between January 1, 1990, and December 31, 1993 were reviewed. Preoperative renal ultrasounds and intravenous pyelograms (IVP) were evaluated for hydronephrosis based on the final diagnosis established by the radiologists. The severity of prolapse was determined from the preoperative office examination or from the examination under anesthesia at the time of surgery. RESULTS: Of 375 patients, 323 had either a preoperative renal ultrasound or IVP. The mean age was 66.0 +/- 10.2 years (range 35-93) and median parity was 3.0 (range 0-10). Of the 323 patients, 25 (7.7%, 95% confidence interval 5, 11) had hydronephrosis. Thirteen patients (4.0%) had mild hydronephrosis, nine (2.8%) had moderate hydronephrosis, and three (0.9%) had severe hydronephrosis. The prevalence of hydronephrosis increased with increasing severity of prolapse. Two patients with hydronephrosis had evidence of renal insufficiency (creatinine > or = 1.6), and both had severe bilateral hydronephrosis and complete procidentia. The prevalence of hydronephrosis was lower in patients with vaginal vault prolapse versus uterine prolapse (3.9% compared with 12.6%, P < .01), CONCLUSION: The prevalence of hydronephrosis in patients undergoing surgery primarily for pelvic organ prolapse is low, increases with worsening pelvic organ prolapse, and is lower in patients with vaginal vault prolapse that in those with uterine prolapse. PMID- 9207808 TI - Pelvic organ prolapse in young women. AB - OBJECTIVE: To determine differences in the characteristics and type of genital prolapse in young women compared with older women. METHODS: A retrospective analysis was performed, identifying 647 women who underwent surgical repair of various types of genital prolapse for the years 1979-1991. One hundred ninety-one patients met our inclusion criteria, having well-documented genital prolapse to or beyond the hymen. Patients were stratified into two age groups, those over 35 years and those 35 or younger. The patients were compared regarding "complexity" of prolapse (ie, the total number of deficient sites per patient), grade of prolapse, parity and coexistent medical conditions. RESULTS: During the study period, 27 young women (mean age +/- standard deviation [SD] 30.3 +/- 3.4 years) and 164 older women (mean age +/- SD 60.6 +/- 11.9 years) met our criteria. Young women were more likely than older women to have 1) potential predisposing medical conditions (congenital anomalies or neurologic or connective tissue diseases) (22.2% versus 6.7%, P < .05), 2) lower mean parity (2.8 versus 3.4, P < .05), 3) only one site of prolapse (56% versus 23%, P < .01), and 4) lower grade of prolapse (33% versus 87% grade 3 or higher, P < .001). CONCLUSION: Young patients undergoing surgery for genital prolapse were more likely to have lower parity and single-site and lower-grade prolapse. A higher than expected prevalence of congenital anomalies, as well as rheumatologic and neurologic diseases in the younger women is intriguing, but further study is necessary before these conditions can be implicated in the genesis of genital prolapse. PMID- 9207810 TI - Comparative morbidity and charges associated with route of hysterectomy and concomitant Burch colposuspension. AB - OBJECTIVE: To compare the surgical morbidity, postoperative course, and hospital charges of Burch colposuspension performed in conjunction with abdominal versus vaginal hysterectomy. METHODS: Power analysis indicated that 35 women would be needed in each group to detect a 20% difference in hospital charges between groups with a beta error of 20% and an alpha error of 5%. A computerized records search identified 80 women who underwent Burch colposuspension, 40 of whom underwent concomitant vaginal hysterectomy (vaginal group) and 40 of whom underwent concomitant abdominal hysterectomy (abdominal group). All procedures were performed by one of 16 surgeons at either Good Samaritan Hospital, Cincinnati, Ohio, or the Medical Center of Central Georgia, Macon, Georgia, between 1992 and 1996. Data on demographics, perioperative course, uterine weight, and operative and total hospital charges were obtained for each group. RESULTS: There was no statistically significant difference in demographics, surgical history, postoperative hemoglobin and hematocrit decrease, postoperative complications (10 versus 23%), operative charges ($4417 +/- 1200 versus $4731 +/- 1453), mean uterine weight (113.5 +/- 45 versus 125.8 +/- 45 g), and operative times (3.0 +/- 0.8 versus 2.9 +/- 0.7 hours) between the vaginal and abdominal groups, respectively. A post hoc power analysis indicated that each group would require 142 patients to achieve statistical significance for postoperative complication rates. The abdominal group had significantly longer hospital stays (3.1 +/- 1.0 versus 2.6 +/- 0.7 days) and higher charges ($7337 +/- 1828 versus $6342 +/- 1123) than the vaginal group. CONCLUSION: When hysterectomy is performed at the time of colposuspension, the vaginal route should be considered seriously when either surgical approach is clinically appropriate. PMID- 9207811 TI - A study of ruptured tubal ectopic pregnancy. AB - OBJECTIVE: Ectopic pregnancy continues to be a leading cause of maternal morbidity and of reduced childbearing potential among women of reproductive age. Because of tubal rupture it is still the main cause of pregnancy-related death during the first trimester. The purpose of our study was to evaluate factors that may predispose a woman to rupture of a tubal ectopic pregnancy. METHODS: In this retrospective study of 693 ectopic pregnancies from three McGill University teaching hospitals, we compared risk factors, preoperative ultrasound, and serum hCG levels between cases with ruptured and unruptured tubal ectopic pregnancy. RESULTS: The age and the number of pregnancies among the two groups of women were similar. The gestational age of women with an unruptured tube was 6.9 +/- 1.9 weeks, and of those with a ruptured tube, the gestational age was 7.2 +/- 2.2 weeks. Tubal rupture was encountered more often in women with at least one child than in childless women. History of ectopic pregnancy was found in 35% of women with an unruptured tubal pregnancy and in 26% of those with a ruptured tube. Serum hCG levels at the time of treatment were not significantly different among the two groups of women. Eleven percent of women with a ruptured tube had serum beta-hCG levels of less that 100 IU/L. CONCLUSION: Tubal rupture is encountered more often in women with no history of ectopic pregnancy and in those with at least one child. This suggests that ectopic pregnancy is less suspected in these women. Tubal rupture is encountered less often in ampullary pregnancy and in small ectopic pregnancies. There is no correlation between serum beta-hCG levels and tubal rupture, and rupture can occur even when serum beta-hCG levels are very low. PMID- 9207812 TI - Over-the-counter and alternative medicines in the treatment of chronic vaginal symptoms. AB - OBJECTIVE: To investigate the use of over-the-counter and alternative medicines in women with chronic vaginal symptoms. METHODS: One hundred five patients, referred by their gynecologists for evaluation of chronic vaginal symptoms, were interviewed about their use of over-the-counter and alternative medicines during the preceding year, the amount of money spent on each, and whether or not their physicians had been informed of these treatments. RESULTS: The mean age was 36 years, and one-half had finished college. The median symptom duration was 2 years. Seventy-seven (73.3%) patients had self-treated with over-the-counter medications such as miconazole (74% of over-the-counter users), clotrimazole (38.2%), or povidone-iodine (13.2%). The median expenditure for over-the-counter use was %0 (range $2-1000). Forty-four (41.9%) had used alternative medicines, most frequently acidophilus pills orally (50%) vaginally (11.4%), yogurt orally (20.5%) or vaginally (18.2%), vinegar douches (13.6%), and boric acid (13.6%). The median expenditure for alternative medicines was $35 (range $0-1200). Fewer physicians were aware of the use of alternative (70.5%) that of over-the-counter medicines (88.3%). Although most patients thought that vulvovaginal candidiasis was the cause of their symptoms, the most common diagnoses at initial presentation were candidiasis in 29 (27.6%), vulvar vestibulitis in 18 (17.1%), irritant dermatitis in 16 (15.2%), and bacterial vaginosis in 11 (10.5%). Women who actually had candidiasis were more likely to have used alternative medicines (odds ration 2.31, 95% confidence interval 1.00, 5.42) than other patients. CONCLUSION: Women with chronic vaginal symptoms often use over-the-counter and alternative medicines that add to health care costs and are unlikely to be of benefit. PMID- 9207813 TI - Endovaginal sonography for the diagnosis of upper genital tract infection. AB - OBJECTIVE: To determine the clinical utility of transvaginal sonography for the diagnosis of upper genital tract infection. METHODS: Fifty-five women who either met the Centers for Disease Control and Prevention's minimal criteria for acute pelvic inflammatory disease or were being seen for non-classic signs of upper genital tract infection were evaluated. During abdominal and endovaginal ultrasound testing, fluid in the cul-de-sac, discrete tubes with or without tubal fluid, multicystic ovaries, and adnexal masses were noted. Upper genital tract infection was confirmed by laparoscopic visualization or histologic or microbiologic evidence of salpingitis of endometritis. RESULTS: The specificity of identifying fallopian tubes with or without intraluminal fluid on ultrasound was 97% (35 of 36); the sensitivity, however, was only 32% (six of 19). Calculated using Bayes theorem and based on a prevalence rate of 50%, the positive predictive value of visualizing fallopian tubes was 91%. The sensitivities associated with the visualization of a multicystic ovary or tubo ovarian abscess were 42% (eight of 19) and 32% (six of 19), with specificities of 86% (31 of 36) and 97%, (35 of 36), and positive predictive values of 75% and 91%, respectively. Cul-de-sac fluid was associated with low sensitivity (37%; seven of 19), low specificity (58%; 21 of 36), and the lowest positive predictive value (47%). CONCLUSION: Endovaginal sonography has limited clinical utility in the diagnosis of upper genital tract infection due to its low sensitivity. PMID- 9207814 TI - Pelvic fluid collections by sonography and febrile morbidity after abdominal hysterectomy. AB - OBJECTIVE: To assess the range of normal findings at endovaginal sonography after abdominal hysterectomy and to assess the relationship these findings and febrile morbidity. METHODS: Fifty-eight women had endovaginal ultrasound at a median of 4 days after abdominal hysterectomy. The volume of fluid in the cul-de-sac and its sonographic characteristics were assessed. Ultrasound findings, which were not released to the patients' physicians, were correlated with febrile morbidity and clinical outcomes. RESULTS: The median pelvic fluid volume was 3.4 mL (interquartile range 0-16.8 mL). No pelvic fluid was detected in 22 of 58 women (37.9%). In the other 36 women, fluid volumes ranged between 0.2 and 76.3 mL. Febrile morbidity was present in 15 of 58 women (26%) overall: eight of 36 (25%) with and seven of 22 (32%) without pelvic fluid. There was no association between the presence of pelvic fluid collections and febrile morbidity (P = .54) or prolonged fever (P = 1.00). There was no difference in the median or mean fluid volumes between women with and without febrile morbidity. The study had a power of 90% with alpha = .05 to detect a difference of 20 mL. Even women with fixed, markedly echoic fluid collections larger than 35 mL did not have significantly more febrile morbidity than women with no pelvic fluid (P = .33). CONCLUSION: The volume of pelvic fluid 3-5 days after hysterectomy does not predict febrile morbidity or the need for drainage. Large or complex fluid collections may be present without adverse clinical consequences, and discovering such a collection in a patient with febrile morbidity after hysterectomy does not necessitate antibiotic therapy or surgical drainage of the fluid collection. PMID- 9207815 TI - The risk of preterm birth across generations. AB - OBJECTIVE: To examine the risk of preterm birth for mothers who themselves were born before term. METHODS: Data were taken from a linked data base of birth certificates composed of two cohorts: 1) a parental cohort of women born between 1947 and 1957 and 2) their offspring born between 1970 and 1992. "Preterm mothers" were women in the parental cohort who were born at less than 37 weeks' gestation. "Term mothers" were women in the parental cohort born at or after 38 weeks' gestation. Preterm mothers and term mothers were matched for birth year, county of birth, marital status, parity, and age. Odds ratios (ORs) and 95% confidence intervals (CIs) were calculated for the risk of preterm delivery in preterm mothers. Multiple logistic regression was used to assess the interaction of concomitant variables with the risk of premature delivery. RESULTS: The risk of preterm birth was significantly higher in preterm mothers than in term mothers (OR 1.18; 95% CI 1.02, 1.37). The risk increased as the gestational age at the mothers' birth decreased (less than 30 weeks'; OR 2.38; 95% CI 1.37, 4.16). The interaction between maternal age and parity increased the risk of preterm delivery at less than 34 weeks in some age and parity strata. CONCLUSION: An increased risk of preterm delivery exists for women who themselves were born before 37 weeks' gestation. The risk is inversely correlated with the maternal gestational age at birth and is influenced by maternal age and parity. PMID- 9207816 TI - Does reducing the number of prenatal office visits for low-risk women result in increased use of other medical services? AB - OBJECTIVE: To determine whether a schedule of fewer prenatal visits than traditional for women with low-risk pregnancies lead to additional medical services outside prescribed prenatal care. METHODS: In a randomized, controlled trial conducted within a group-model health maintenance organization, we studied 2328 pregnant women judged to be a low risk of adverse perinatal outcomes. After risk assessment and consent, women were assigned to an experimental (nine visits) or a control (14 visits) schedule, with additional visits if requested either by providers after identifying risks or by women seeking additional services. We recorded whether women underwent maternal serum alpha-fetoprotein screening, obstetric ultrasound examinations at 15-24 weeks' gestation, hematocrit testing after 20 weeks, and diabetic screening. We also noted visits to nonobstetric care providers or our emergency care center, telephone calls, and hospitalizations. RESULTS: We found no significant differences between the two groups for prenatal blood tests, visits to nonobstetric providers or to the emergency care center, telephone calls from patients, or hospital admissions. A significantly greater percentage of women underwent ultrasound examinations at 15-24 weeks in the control group compared with the experimental group (57.3% and 53.1%, respectively; P = .045). CONCLUSION: The reduction in prenatal visits achieved using the experimental schedule was not accompanied by an increase in the use of other medical services compared with the routine schedule. The use of the schedule proposed by the Expert Panel on the Content of Prenatal Care improved the efficiency of delivery of prenatal care to low-risk women. PMID- 9207817 TI - Patterns of prenatal care initiation in Georgia, 1980-1992. AB - OBJECTIVE: To determine whether characteristics in a woman's first pregnancy were associated with the trimester in which she initiated prenatal care in her second pregnancy. METHODS: Data for white and black women whose first and second pregnancies resulted in singleton live births between 1980 and 1992 were obtained from Georgia birth certificates (n = 177,041). Adjusted relative risks (RRs) for early prenatal care in the second pregnancy were computed by logistic regression models that included trimester of prenatal care initiation, infant outcomes, or maternal conditions in the woman's first pregnancy as the exposure and controlled for maternal age, education, child's year of birth, interval between first and second pregnancy, presence of father's name on the birth certificate, and the interaction between prenatal care and education. Models were stratified by race. RESULTS: Women of both races who initiated prenatal care in the first trimester of their first pregnancies were more likely than those with delayed care to initiate prenatal care in the first trimester of their second pregnancies (RR = 1.25 and 1.63 for white and black women educated beyond high school, respectively). Both white and black women who delivered a baby with very low birth weight (RR = 1.06 and 1.15, respectively) or who suffered an infant death (RR = 1.09 and 1.31, respectively) in their first pregnancies were more likely than those who did not experience these events to begin prenatal care in the first trimester of their second pregnancies. CONCLUSION: Women with some potentially preventable adverse infant outcomes tend to obtain earlier care in their next pregnancy. Unfortunately, women who delayed prenatal care in their first pregnancy frequently delay prenatal care in their next. PMID- 9207818 TI - Underreporting of maternal mortality in The Netherlands. AB - OBJECTIVE: To establish the actual number of maternal deaths in The Netherlands by determining the degree of underreporting. METHODS: We conducted a nationwide, retrospective cross-check of the three available maternal mortality registration systems and issued a questionnaire to senior obstetricians in all hospitals during the years 1983-1992. RESULTS: The officially reported maternal mortality rate during the study period was 7.1 per 100,000 live births (133 maternal deaths per 1,862,985 live births). After completion of the study, our data indicate that the rate should be at least 9.7 per 100,000 live births (180 maternal deaths). Early pregnancy and indirect deaths were more likely to be underreported than direct deaths during labor and the puerperium. Failure to register the recent pregnancy on the death certificate was a frequent problem. Misclassification was particularly evident for cerebrovascular disorders, cardiovascular disorders, and eclampsia. CONCLUSION: The level of underreporting of maternal mortality in The Netherlands was estimated at 26%. The pregnancy status of women should be registered on death certificates. Officially reported maternal mortality rates are unreliable and international comparisons using these data thus are less meaningful. PMID- 9207819 TI - Double-blind, placebo-controlled study of ranitidine for gastroesophageal reflux symptoms during pregnancy. AB - OBJECTIVE: To determine whether ranitidine (Zantac) taken once or twice daily is effective for relieving symptoms of gastroesophageal reflux among pregnant women who had failed conservative measures. METHODS: Volunteers with heartburn despite antacids were sought among our obstetrics clinic population for this double blind, placebo-controlled, triple crossover trial. After a baseline week, 20 patients were randomized to receive the three following weekly regimens: ranitidine 150 mg twice daily, placebo in the morning and ranitidine 150 mg in the evening, or placebo twice daily. Daily scores on symptom diaries, global assessments, and number of antacids taken were compared among the 18 patients completing the study. RESULTS: The twice-daily dosage of ranitidine was the only regimen found to reduce heartburn symptoms when compared with the baseline (P < .001) or a placebo (P < .01). Compared with ranitidine taken once daily, the twice-daily dosing prompted less need for antacid tablets compared with the placebo (P < .05 versus P > .05) and to the baseline (P < .001 versus P < .05). The average reduction of heartburn severity using twice-daily ranitidine was 55.6% when compared with baseline (95% confidence interval [CI] 34.8%, 76.5%) was 44.2% when compared with placebo (95% CI 15.4%, 72.9%). CONCLUSION: This study indicates the efficacy of ranitidine 150 mg taken twice daily, rather than once daily, for relief of gastroesophageal reflux symptoms during pregnancy. PMID- 9207820 TI - Absorption kinetics of misoprostol with oral or vaginal administration. AB - OBJECTIVE: To compare the pharmacokinetics of vaginal and oral administration of the prostaglandin E1 analogue, misoprostol. METHODS: Twenty women received 400 micrograms doses of misoprostol either orally or as tablets placed in the vagina. Serum levels of principal metabolite, misoprostol acid, were measured at 7.5, 15, 30, 45, 60, 90, 120, and 240 minutes. The first ten women were pregnant and undergoing first-trimester abortions, and the last ten were not pregnant and had additional blood sampling at 360 minutes. We compared the pharmacokinetics of misoprostol acid after oral and vaginal administration. RESULTS: All 20 subjects completed the study. The maximum mean (+/- standard deviation [SD]) of misoprostol acid differed significantly between the oral and vaginal groups (277 +/- 124 compared with 165 +/d- 86 pg/mL, respectively; P = .03, analysis of variance), as did the mean +/- SD time to peak levels (34 +/- 17 compared with 80 +/- 27 minutes, respectively; P < .001) and areas under the misoprostol concentration versus time curve (mean +/- SD) up to 4 hours (n = 20,273.3 +/- 110.0 compared with 503.3 +/- 296.7 pg.hour/mL, respectively; P = .033) and up to 6 hours (n = 10, 300.0 +/- 103.3 compared with 956.7 +/- 541.7 pg.hour/mL, respectively; P = .029). The extent of absorption was highly variable among subjects in each group. CONCLUSION: There are significant differences in the pharmacokinetics of misoprostol administered by vaginal and oral routes that may explain the difference observed in clinical efficacy. Assuming that the pharmacologic effect of misoprostol is related to its concentration in the plasma, our observation of the prolonged serum concentrations in the vaginal group suggests that vaginal administration could be dosed at longer intervals than oral. PMID- 9207821 TI - The developmental outcome of children with antenatal mild isolated ventriculomegaly. AB - OBJECTIVE: To evaluate standardized developmental test performance of infants and children who as fetuses had mild isolated cerebral ventriculomegaly diagnosed by ultrasound. METHODS: Ultrasound records from 1990 to 1996 were searched for cases of mild isolated ventriculomegaly, and standardized developmental testing of the children was offered to their parents. Each consented child was matched to a normal antepartum subject with respect to sex, race, indication for ultrasound, and gestational age (+/- 2 weeks) at the time of ultrasound. Tests of cognitive, motor, and adaptive behavior were then administered by examiners blinded to the subjects' case or comparison status. RESULTS: Twenty-two cases and an equal number of matched comparison subjects completed the testing. The ventriculomegaly and comparison groups were similar with respect to parental age, maternal education, and household income. The ventriculomegaly subjects scored significantly lower than the comparison group on both the Bayley Scales of Infant Development: mental development index (88.95 versus 99.68, P = .017) and psychomotor development index (95.99 versus 103.95, P = .039). Eight of the 22 ventriculomegaly children were classified as developmentally delayed on the mental developmental index compared with one of 22 children in the comparison group (P = .021). Adaptive behavior skills, as measured by the Vineland Behavior Scales (99.64 versus 102.68), were not significantly different between the groups (P = .571). CONCLUSION: Mild isolated ventriculomegaly detected on antepartum sonographic examination is associated with a significant risk for developmental delay. Insofar as these children were judged to be completely normal at birth, our findings represent an important application of antepartum sonography for identifying infants who could be targeted for early childhood intervention. PMID- 9207822 TI - Serum triple-marker screening in in vitro fertilization and naturally conceived pregnancies. AB - OBJECTIVE: To determine whether results of second-trimester maternal serum triple marker screening for Down syndrome and open neural tube defects in singleton pregnancies conceived from in vitro fertilization (IVF) differ from those of pregnancies conceived spontaneously. METHODS: The screen-positive rates and triple-marker levels of patients conceiving singleton pregnancies by IVF were compared to age-adjusted standards. RESULTS: Sixty-nine singleton IVF pregnancies with maternal serum screening were identified. Twenty-one (30.4%) of the 69 IVF singleton pregnancies had a positive screen for Down syndrome compared with a 14.4% expected screen-positive rate for the maternal age distribution in our observed sample (P = .013). The screen-positive rate for open neural tube defects in the measured population was similar to anticipated values based on historic controls (5.8% in IVF patients versus 5.3% in the total population). The median levels of the triple markers were 0.95 multiples of the median (MoM) for alpha fetoprotein (AFP), 0.90 MoM for unconjugated estriol (E3), and 1.22 MoM for hCG. CONCLUSION: The increased hCG levels as well as the slightly lower AFP and unconjugated E3 levels may contribute to the higher Down syndrome screen-positive rate in this IVF singleton population. These results may be due to the number of embryos transferred, the maternal hormonal environment of the IVF process, or other factors. Pregnancies conceived by IVF may be twice as likely to have a positive maternal serum screening test. As additional data are collected, corrected standards should be determined. PMID- 9207823 TI - Electrical activity of the human uterus during pregnancy as recorded from the abdominal surface. AB - OBJECTIVE: To validate the possibility that human uterine electrical events (electromyographic signals) can be recorded and characterized from the abdominal surface during pregnancy. METHODS: The gestational ages ranged from 20 to 43 weeks. The study included patients at term but not in labor, patients in active labor (term and preterm), postpartum patients, and patients followed monthly during their pregnancy (n = 40). Uterine electrical activity in the frequency range of 0.3-50 Hz was recorded using bipolar electrodes placed on the abdominal surface. In some patients, intrauterine pressure also was measured. Power spectral analysis was performed using the fast Fourier transform. RESULTS: Throughout most of pregnancy, uterine electrical activity was minimal, consisting of infrequent and low-amplitude electromyographic bursts. When bursts occurred before labor, they often corresponded to perceptions of contractility by the patient. During term and preterm labor, bursts of electromyographic activity were frequent and of large amplitude and were correlated with large transient changes in the intrauterine pressure and with pain. Fast Fourier transform analysis of the bursts during active term labor demonstrated a peak frequency of 0.71 +/- 0.05 Hz, compared with 0.48 +/d- 0.03 Hz before labor. Spectral analysis also showed a fivefold increase in the peak energy levels of the bursts during term labor (60.2 +/- 13.87 mu Vs) and preterm labor (62.3 +/- 22.93 mu Vs) compared with earlier in gestation (11.36 +/- 4.03 mu Vs at 27-36 weeks; P < .05). CONCLUSION: Recording of uterine electromyographic activity from the abdominal surface is a reliable method to follow the evolution of uterine contractility during pregnancy and during term and preterm labor. Further studies will define the usefulness of this noninvasive technology in the prediction and management of labor. PMID- 9207824 TI - Placental cavities. AB - OBJECTIVE: To study placental cavities by gross and microscopic examination and ultrasonography and their frequency with various epidemiologic factors and intervillous thrombosis. METHODS: After formalin fixation, interval sections of 567 placentas were prepared to search for cavities and intervillous thrombosis. Cavities were subjected to histologic and ultrasonographic examinations. RESULTS: Frequency of cavities with diameter of 1 cm or more was 34.9% in 567 mature placentas. Frequency of cavities was significantly higher in heavy, thick placentas associated with male fetuses. Histologic examination revealed villus laceration in cavities and syncytial cells, isolated chorionic villi, or air bubbles in placental fetal veins. All 82 placentas with cavities showed villus lacerations in the cavities and air bubbles in the fetal veins. Intervillous thromboses in fetal lobules were located only in the cavities. Cavities were first found by ultrasonography at a mean gestational age of 30.9 +/- 3.8 weeks. Ultrasonography did not always differentiate accurately between intervillous thrombosis and cavities. CONCLUSION: Placental cavities were found significantly more often in heavy, thick placentas associated with male fetuses. Strong uterine contractions during placental detachment could produce villus laceration in cavities, following contamination by air bubbles and isolated villus tissue in the fetal veins. Placental cavities are vulnerable to villus laceration. Intervillous thrombosis occurred only in cavities. PMID- 9207825 TI - Trophoblastic apoptosis in mice with preterm delivery and its suppression by urinary trypsin inhibitor. AB - OBJECTIVE: To investigate histopathologic changes of the placenta in mice with preterm delivery induced by lipopolysaccharide and the effect of urinary trypsin inhibitor. METHODS: Female C3H/HeN mice impregnated by male B6D2F1 mice were treated with lipopolysaccharide (50 micrograms/kg, intraperitoneally) or lipopolysaccharide plus urinary trypsin inhibitor (25 x 10(4) U/kg, intraperitoneally). At 3, 6, 9, and 18 hours after the second dose of lipopolysaccharide, and at delivery in the control and urinary trypsin inhibitor treated groups, the concentrations, of interleukin-1 alpha and tumor necrosis factor-alpha were determined in serum and amniotic fluid. Subsequently, the placentas were examined. In the same manner, we examined mice treated with interleukin-1 alpha (250 micrograms/kg, subcutaneously) on day 15 of pregnancy and intact mice on days 15 and 18 of pregnancy as well as at delivery. To assess the direct action of cytokines, we cultured placental slices with tumor necrosis factor-alpha, interleukin-1 alpha, or tumor necrosis factor-alpha plus urinary trypsin inhibitor and examined them morphologically. RESULTS: Control mice were characterized by trophoblastic apoptosis and increased serum levels of tumor necrosis factor-alpha and interleukin-1 alpha. In contrast, urinary trypsin inhibitor-treated mice showed suppression of apoptosis and lower cytokine levels. Interleukin-1 alpha induced trophoblastic apoptosis and increased the serum level of tumor necrosis factor-alpha. The in vitro study showed that tumor necrosis factor-alpha directly induced trophoblastic apoptosis in placental slices. CONCLUSION: We demonstrated that trophoblastic apoptosis occurs in the placentas of a mouse model with preterm delivery induced by lipopolysaccharide. We postulated that apoptosis may lead to placental abruption, and its development may be prevented by treatment with urinary trypsin inhibitor. PMID- 9207826 TI - Risk factors for acidemia at birth. AB - OBJECTIVE: To identify risk factors for acidemia at birth. METHODS: From September 1988 to December 1996, cord arterial blood pH was measured in 23,016 of 27,064 live-born infants (85.0%). Values below 7.05 were observed in 264 newborns (1.1%), of whom 14 born by cesarean delivery before labor and one triplet infant were excluded from the study. The remaining 249 newborns with acidemia and their mothers were compared with 249 unmatched controls with normal pH (the first infant with umbilical arterial pH above 7.10 born after each case). Multivariate logistic regression was used to adjust for potentially confounding variables. RESULTS: Variables significantly and independently associated with acidemia at birth were labor with breech presentation (adjusted odds ratio [OR]2.9), oxytocin administration (OR 2.1), meperidine administration (OR 2.0), cord entanglement (OR 1.7), and male gender of the infant (OR 1.4). Clinical evidence of chorioamnionitis also was associated with acidemia, although after adjustment for prematurity, the association was not statistically significant (OR 3.9, 95% confidence interval 0.8, 19). CONCLUSION: Labor with breech presentation, administration of oxytocin and meperidine, cord entanglement, and male gender are associated with an increased risk for insufficient fetomaternal gas exchange. PMID- 9207827 TI - Higher rate of fetal acidemia after regional anesthesia for elective cesarean delivery. AB - OBJECTIVE: To determine the prevalence of fetal acidemia associated with regional anesthesia for elective cesarean delivery in healthy paturients with uncomplicated singleton term pregnancies. METHODS: This was an epidemiologic study using the data base of the Swiss obstetric study group (Arbeitsgemeinschaft Schweizerischer Frauenkliniken). After the exclusion of cases with extraneous factors that may have affected the health of the neonate, we analyzed the umbilical artery pH, Apgar score, and other neonatal outcome measures after cesarean delivery with reference to the anesthetic technique. RESULTS: From 1985 to 1994, 327,763 deliveries, including 40,858 (12.47%) by cesarean, were registered in the data base. Of these, 5806 patients fulfilled the study criteria. The study population included 1002 spinal, 2155 epidural, and 2649 cases of general anesthesia. The frequency of fetal acidemia (pH less than 7.10) was significantly increased in the spinal-anesthesia group (odds ratio [OR] 4.67; 95% confidence interval [CI] 2.73, 8.01) and in the epidural group (OR 2.39; 95% CI 1.42, 4.04) compared with the general-anesthesia group. CONCLUSION: The rate of fetal acidemia is significantly increased after regional anesthesia. This risk must be judged in light of the risks inherent with general anesthesia. PMID- 9207828 TI - The effect of instituting an elective labor epidural program on the operative delivery rate. AB - OBJECTIVE: To evaluate labor outcome as well as maternal and neonatal morbidity before and after the initiation of elective labor epidural capability. METHODS: On October 1, 1993, a sudden change in military requirements mandated provision of elective labor epidural capability at our institution. Before this time, epidural provision had been primarily in response to urgent obstetrician requests. Pre-labor and labor characteristics and outcomes were reviewed for the year before this policy change (group 1, n = 373) and for the year after it (group 2, n = 421) in a population of nulliparous patients delivering singleton, vertex fetuses at 36-42 weeks' gestational age. In addition, the group of patients receiving labor epidurals before their ready availability (group 1E, n = 49) was compared with the group receiving them after ready availability (group 2E, n = 247). RESULTS: Maternal labor characteristics showed a slight (10 minutes on average) prolongation of the second stage of labor in group 2. The incidence of diagnosed chorioamnionitis was higher in group 2. Patients receiving epidurals in each time frame were analyzed to identify epidural-related findings, as opposed to findings associated with intrinsically more problematic labors. Epidural-related factors included the slightly prolonged second stage of labor, increased use of oxytocin, and a higher incidence of diagnosed chorioamnionitis. CONCLUSION: Our study demonstrated no increase in the rate of operative deliveries in a population that suddenly received access to on-request labor epidurals. We believe this option should be offered to parturients without making them feel that they must choose between comfort and safety. PMID- 9207829 TI - Psychosocial risk factors associated with cocaine use during pregnancy: a case control study. AB - OBJECTIVE: To investigate whether psychosocial risk factors predict cocaine use during pregnancy. METHODS: We sampled 229 pregnant women from an urban prenatal clinic. Drug use ascertainment was based on self-report and urine toxicology, with 102 subjects classified as drug users and 127 as nonusers. A questionnaire measuring seven psychosocial risk factors was administered. The predictive relation between these characteristics and drug use was ascertained through multivariate analyses, controlling for potential sociodemographic confounders. RESULTS: Six of the seven psychosocial risk factors were significant predictors for this sociodemographic group. Women who used cocaine during pregnancy were more likely to have a family history of alcohol or drug problems, to have been introduced to drugs by a male partner, to be depressed, to have less social support, to have current partners who were substance users, and to have less stable living situations. Both groups of subjects had high rates of childhood sexual abuse, but this alone was not predictive of drug use. In addition, cigarette smoking was a strong predictor of illicit drug use. CONCLUSION: Identification of multiple psychosocial risk factors has implications for the identification and treatment of substance-using pregnant women. Because cocaine users and nonusers did not differ in gestational age at entry into prenatal care, opportunities exist for intervention during pregnancy. Based on the study findings, evaluation of the following aspects of a patient's lifestyle can aid in the detection of cocaine use during pregnancy: smoking status, family history of alcohol or drug problems, and current drug use by a male partner. PMID- 9207830 TI - Development of a computerized telecommunication system for in-training evaluation of residents in a laparoscopic educational program. AB - OBJECTIVE: To describe our initial experience with a computerized telecommunication system, termed the interactive voice-response system, to record resident performance of laparoscopic surgery. METHODS: After completing a laparoscopic procedure, the surgeon and resident telephone a toll-free number independently and respond to three prerecorded statements using a Likert scale of 1 to 5. The caller then is asked to describe the resident's response to critical incidents or elements of surprise that arose during the surgery. The ratings and verbal comments are compiled, transcribed, and forwarded to the respective resident. The resident (and program director) can hear the verbal comments by entering a four-digit code. RESULTS: Between May 1, 1995, and May 31, 1996, 430 cases were reported by 11 surgeons and 16 residents using the interactive voice response system. One hundred ninety-five (45%) procedures were entered by both the resident and surgeon. A survey undertaken during the introductory phase of the project revealed that five of the seven residents exposed to the system found that it provided useful feedback and preferred the system to traditional in service reporting methods. In addition, five residents thought that the system complemented the personal feedback they received in the operating room. CONCLUSION: The system has been accepted by both residents and surgeons and has addressed the important components of resident in-training evaluation, namely, evaluation on a case-by-case basis, timely feedback, and self-assessment of resident performance. PMID- 9207831 TI - Persistent intraepithelial neoplasia after excision for cervical intraepithelial neoplasia grade III. PMID- 9207832 TI - Oral administration of misoprostol for labor induction: a randomized controlled trial. PMID- 9207833 TI - A randomized trial of misoprostol and oxytocin for induction of labor: safety and efficacy. PMID- 9207834 TI - Transcription factor phosphorylation by a protein kinase associated with chloroplast RNA polymerase from mustard (Sinapis alba). AB - The chloroplast transcription machinery involves multiple components with both catalytic and regulatory functions. Here we describe a serine-specific protein kinase activity that is associated with the major chloroplast RNA polymerase and phosphorylates sigma-like transcription factors in vitro. The kinase activity can be assigned to a 54 kDa polypeptide of partially purified RNA polymerase (KPC, kinase polymerase complex). This polypeptide is also present in a smaller complex that contains several putative polymerase subunits and reveals kinase activity but lacks transcription activity (KC, kinase complex). Although the 54 kDa component could not be chromatographically separated from the rest of this complex without loss of activity, it retained residual kinase activity in an electrophoretic blot assay. The polymerase-associated kinase is itself affected by in vitro phosphorylation and dephosphorylation, which raises the possibility that it is part of a signalling cascade that controls chloroplast transcription in vivo by factor phosphorylation. PMID- 9207835 TI - The RY sequence is necessary but not sufficient for the transcription activation of a winged bean chymotrypsin inhibitor gene in developing seeds. AB - A winged bean Kunitz-type chymotrypsin inhibitor (WCI) is expressed in seeds and tuberous roots. In seeds, the expression of WCI is restricted to the period between the mid- and late-maturation stage. To understand the mechanisms that regulate the expression of WCI genes, we analyzed the promoter activity of the upstream region of the WCI-3b gene, which encodes a major WCI protein, in transgenic tobacco plants. By using a series of constructs with 5' deletions in the upstream sequences, the region between -882 and -623, relative to the transcription start site, was shown to contain multiple sequences which are responsible for high level expression in mid-maturation stage seeds. However, when this region was fused to the cauliflower mosaic virus 35S core promoter in both orientations, the chimeric promoters showed only a weak transcription activity in transgenic tobacco plants. Further analyses using internal deletion constructs revealed that the region between -882 and -174 is required for the transcription activation. Disruption of the RY sequence at -517, which is conserved in many seed protein genes, resulted in a drastic reduction of the transcription activity in seeds. These results suggest that sequences necessary for high level induction of the WCI-3b gene transcription in developing seeds are dispersed in the region between -882 and -174, and that the RY sequence is one of these sequences. PMID- 9207836 TI - A novel gene whose expression in Medicago truncatula roots is suppressed in response to colonization by vesicular-arbuscular mycorrhizal (VAM) fungi and to phosphate nutrition. AB - A cDNA clone (Mt4) was isolated as a result of a differential screen to identify genes showing altered expression during the interaction between Medicago truncatula and the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus versiforme. Mt4 represents a M. truncatula mRNA that contains numerous short open reading frames, the two longest of which are predicted to encode polypeptides of 51 amino acids each. One of these open reading frames shares a short region of identity with a phosphate starvation-inducible gene from tomato. Mt4 gene expression is regulated in response to colonization by mycorrhizal fungi: transcripts were detected in non-colonized roots and levels decreased in both M. truncatula and M. sativa (alfalfa) roots after colonization by G. versiforme. Transcript levels also decreased during the incomplete interaction between G. versiforme and a M. sativa mycorrhizal minus (myc-) line, indicating that the down-regulation of this gene occurs early during the interaction between the fungus and its host plant. Phosphate levels in the nutrient media also affected the expression of the Mt4 gene: transcripts were present in the roots of plants grown under phosphate-deficient conditions, but were undetectable in the roots of plants grown under phosphate sufficient conditions. Furthermore, expression was only observed when plants were grown under nitrogen-sufficient conditions. Northern blot analyses indicate that Mt4 transcripts are present primarily in roots and barely detectable in stems or leaves. Thus, Mt4 represents a M. truncatula gene whose expression is regulated in response to both colonization by mycorrhizal fungi and to the phosphate status of the plant. PMID- 9207837 TI - Characterization of nuclease activities and DNA fragmentation induced upon hypersensitive response cell death and mechanical stress. AB - Programmed cell death (PCD) is activated during the response of multicellular organisms to some invading pathogens. One of the key aspects of this process is the degradation of nuclear DNA which is thought to facilitate the recycling of DNA from dead cells. The PCD of tobacco plants (genotype NN) infected with tobacco mosaic virus (TMV) is accompanied by the induction of nuclease activities and the cleavage of nuclear DNA to fragments of about 50 kb. We examined the correlation between the increase in nuclease activities and the fragmentation of nuclear DNA during TMV- and bacteria-induced PCD in tobacco. We found that the increase in nuclease activities did not always correlate with fragmentation of nuclear DNA. Thus, in addition to pathogens that induce PCD, mechanical injury and infiltration of leaves with 1 M sucrose or bacteria that did not induce PCD also resulted in an increase in nuclease activities. Analysis of nuclease activities in total leaf extracts, nuclear extracts, and intercellular fluid (i.e., apoplast) revealed that at least four different nuclease activities are induced during PCD in tobacco; of these at least three appear to be secreted into the intercellular fluid. Although the latter were also induced in response to treatments that did not result in DNA fragmentation, they may function in the recycling of plant DNA during late stages of PCD when the integrity of the plasma membrane is compromised. This suggestion is supported by the finding that DNA degradation occurred late during TMV-induced PCD in tobacco. In addition, the finding of induced nuclease activities in the intercellular fluid raises the possibility that they may serve a protective function by degrading the DNA of invading pathogens. PMID- 9207838 TI - Self-incompatibility in the grasses: evolutionary relationship of the S gene from Phalaris coerulescens to homologous sequences in other grasses. AB - Self-incompatibility is widespread in the grasses and it is proposed that the grasses share a common incompatibility mechanism that is distinct from those operating in the dicotyledonous species studied in great detail. Where good genetic data are available, all grass species appear to have an incompatibility mechanism controlled by two unlinked loci, S and Z. A putative S gene has been cloned from Phalaris coerulescens. This gene is characterized by two major domains: an allele specificity domain and a thioredoxin catalytic domain. A family of sequences with varying degrees of homology to this gene has been identified among 15 grass species covering all subfamilies of the Poaceae. These S-related sequences appear to be present in the grass family regardless of self compatibility. Evidence is presented to show that at least one of the sequences is transcribed, suggesting a functional gene. In contrast to the high expression of the S gene in Phalaris pollen, expression of the related gene in the pollen (or anthers) of the grass species examined was so low that RNA gel blot analysis failed to display a significant signal. However, reverse transcription-based polymerase chain reaction (RT-PCR) successfully amplified the region corresponding to the S thioredoxin domain from 10 of the grass species. With grasses other than Phalaris, RT-PCR showed limited success in amplifying the region corresponding to the S variable portion at the 5' end of the Phalaris S gene. Sequencing of the PCR-amplified S thioredoxin region from wheat, barley, rye and Dactylis revealed that this is a highly conserved gene with 94-97% sequence similarity with the corresponding Phalaris S gene. The conservation of sequence and ubiquitous expression of the gene across the grass family strongly suggest that the S-related gene is carrying out a significant biological function in the Poaceae. On the basis of these findings, a model for the evolution of the S self-incompatibility gene in the grasses is proposed. PMID- 9207839 TI - Molecular characterization of an Arabidopsis thaliana cDNA coding for a monofunctional aspartate kinase. AB - A cDNA clone encoding a monofunctional aspartate kinase (AK, ATP:L-aspartate 4 phosphotransferase, EC 2.7. 2.4) has been isolated from an Arabidopsis thaliana cell suspension cDNA library using a homologous PCR fragment as hybridizing probe. Amplification of the PCR fragment was done using a degenerate primer designed from a conserved region between bacterial monofunctional AK sequences and a primer identical to a region of the A. thaliana bifunctional aspartate kinase-homoserine dehydrogenase (AK-HSDH). By comparing the deduced amino acid sequence of the fragment with the bacterial and yeast corresponding gene products, the highest identity score was found with the Escherichia coli AKIII enzyme that is feedback-inhibited by lysine (encoded by lysC). The absence of HSDH-encoding sequence at the COOH end of the peptide further implies that this new cDNA is a plant lysC homologue. The presence of two homologous genes in A. thaliana is supported by PCR product sequences, Southern blot analysis and by the independent cloning of the corresponding second cDNA (see Tang et al., Plant Molecular Biology 34, pp. 287-294 [this issue]). This work is the first report of cloning a plant putative lysine-sensitive monofunctional AK cDNA. The presence of at least two genes is discussed in relation to possible different physiological roles of their respective product. PMID- 9207840 TI - The isolation of a novel metallothionein-related cDNA expressed in somatic and zygotic embryos of Douglas-fir: regulation by ABA, osmoticum, and metal ions. AB - To isolate genes which are expressed preferentially during embryogenesis, a Douglas-fir embryogenesis cDNA library was constructed and differentially screened with cDNA probes made with mRNA from developing and mature embryos, respectively. The cDNA clone PM 2.1 was isolated based on its abundance in developing seeds and absence in mature seeds, and its predicted amino acid sequence was shown to have structural features characteristic of plant MT-like proteins. Alignment of the PM 2.1 predicted amino acid sequence with other plant MT-like protein sequences revealed a general paucity of Cys and Cys-Xaa-Cys sequences and the presence of novel serine residues within the conserved Cys-Xaa Cys motifs in the C-terminal domain. The consensus sequence following the Cys poor spacer in type 2 MT-like proteins, CXCXXXCXCXXCXCX, was modified in PM 2.1 to CXSXXXSXYXX-XCX. Phylogenetic analysis supported PM 2.1 was distinct from other MT and grouped with MT-like proteins from Arabidopsis (OEST), rice (AEST) and kiwifruit (AD1), which do not belong to type 1 or 2. The PM 2.1 gene was expressed in somatic and zygotic embryos, in haploid maternal tissue, as well as in hormone- and metal-treated seeds and seedlings. The PM 2.1 transcripts were detected in the needles of 14-week-old seedlings, but not the root tissue or mature pollen. The expression of the PM 2.1 gene in embryos was dependent upon ABA and osmoticum and in seedlings was differentially modulated by metals, suggesting a role of the PM 2.1 gene product in the control of microelement availability during Douglas-fir seed development and germination. The novel structural features, and the developmental, hormonal and metal modulation of PM 2.1 expression, are evidence for a new type of MT-related protein in plants. PMID- 9207841 TI - A copia-like retrotransposon Tgmr closely linked to the Rps1-k allele that confers race-specific resistance of soybean to Phytophthora sojae. AB - We have isolated and characterized Tgmr, a copia-like retrotransposon, linked tightly to the Rps1-k allele that confers race-specific resistance of soybean to the the fungal pathogen Phytophthora sojae. Southern analysis followed by PCR and sequence analyses, using primers based on sequences flanking the insertion site confirmed that the element was inserted in the neighboring region of Rps1-k but not in that of the other four Rps1 alleles. This implies that Tgmr was transposed into the Rps1-k flanking site after the divergence of Rps1 alleles. Southern analysis of a series of diverse soybean cultivars revealed a high level of polymorphism of Tgmr-related sequences. These results indicate that this low copy retroelement family could have been active in the soybean genome in the recent past. Tgmr contains long terminal repeats (LTR) and four non-overlapping open reading frames (ORF), presumably originating from mutations leading to stop codons of a single ORF. The conserved domains for gag, protease, integrase, reverse transcriptase and RNaseH are present in the internal portion of the element. However, the protease, reverse transcriptase and RNaseH of this element are non-functional due to the presence of several stop codons. Possible transactivation of Tgmr and application of this element in insertional mutagenesis for soybean are discussed. PMID- 9207842 TI - PCR-identification of a Nicotiana plumbaginifolia cDNA homologous to the high affinity nitrate transporters of the crnA family. AB - A family of high-affinity nitrate transporters has been identified in Aspergillus nidulans and Chlamydomonas reinhardtii, and recently homologues of this family have been cloned from a higher plant (barley). Based on six of the peptide sequences most strongly conserved between the barley and C. reinhardtii polypeptides, a set of degenerate primers was designed to permit amplification of the corresponding genes from other plant species. The utility of these primers was demonstrated by RT-PCR with cDNA made from poly(A)+ RNA from barley, C. reinhardtii and Nicotiana plumbaginifolia. A PCR fragment amplified from N. plumbaginifolia was used as probe to isolate a full-length cDNA clone which encodes a protein, NRT2;1Np, that is closely related to the previously isolated crnA homologue from barley. Genomic Southern blots indicated that there are only 1 or 2 members of the Nrt2 gene family in N. plumbaginifolia. Northern blotting showed that the Nrt2 transcripts are most strongly expressed in roots. The effects of external treatments with different N sources showed that the regulation of the Nrt2 gene(s) is very similar to that reported for nitrate reductase and nitrite reductase genes: their expression was strongly induced by nitrate but was repressed when reduced forms of N were supplied to the roots. PMID- 9207844 TI - Cloning and expression of an Arabidopsis thaliana cDNA encoding a monofunctional aspartate kinase homologous to the lysine-sensitive enzyme of Escherichia coli. AB - As in many bacterial species, the first enzymatic reaction of the aspartate family pathway in plants is mediated by several isozymes of aspartate kinase (AK) that are subject to feedback inhibition by the end-product amino acids lysine or threonine. So far, only cDNAs and genes encoding threonine-sensitive AKs have been cloned from plants. These were all shown to encode polypeptides containing two linked activities, namely AK and homoserine dehydrogenase (HSD), similar to the Escherichia coli thrA gene encoding a threonine-sensitive bifunctional AK/HSD isozyme. In the present report, we describe the cloning of a new Arabidopsis thaliana cDNA that is relatively highly homologous to the E. coli lysC gene encoding the lysine-sensitive AK isozyme. Moreover, similar to the bacterial lysine-sensitive AK, the polypeptide encoded by the present cDNA is monofunctional and does not contain and HSD domain. These observations imply that our cloned cDNA encodes a lysine-sensitive AK. Southern blot hybridization detected a single gene highly homologous to the present cDNA, plus an additional much less homologous gene. This was confirmed by the independent cloning of an additional Arabidopsis cDNA encoding a lysine-sensitive AK (see accompanying paper). Northern blot analysis suggested that the gene encoding this monofunctional AK cDNA is abundantly expressed in most if not all tissues of Arabidopsis. PMID- 9207843 TI - Differential induction of seven 1-aminocyclopropane-1-carboxylate synthase genes by elicitor in suspension cultures of tomato (Lycopersicon esculentum). AB - The key enzyme of ethylene biosynthesis, ACC synthase, is encoded by a multigene family. We describe three new DNA sequences encoding members of the ACC synthase family of the tomato. One of these sequences encodes a novel ACC synthase, LE ACS6, which is phylogenetically related to the ACC synthases LE-ACS1A and LE ACS1B. Gene-specific probes for seven tomato ACC synthase genes were prepared. They were used for RNase protection assays to study the accumulation of ACC synthase transcripts in suspension-cultured tomato cells after the addition of an elicitor. The ACC synthase genes LE-ACS2, LE-ACS5 and LE-ACS6 were strongly induced by the elicitor. In contrast, the genes LE-ACS1B, LE-ACS3 and LE-ACS4 were constitutively expressed and LE-ACS1B was present at all times at a particularly high level. Thus, there are two groups of ACC synthase transcripts expressed in these cells, either elicitor-induced or constitutive. A transcript of LE-ACS1A was not detected. Despite the presence of LE-ACS1B, LE-ACS2, LE-ACS3, LE-ACS4 and LE-ACS5, there was only little ethylene produced in the absence of the elicitor. Increased ethylene production is usually correlated with the accumulation of ACC synthase transcripts, indicating that ethylene production is controlled via the transcriptional activation of ACC synthase genes. However, the abundance of several ACC synthase mRNAs studied was not strictly correlated with the rate of elicitor-induced ethylene production. Our data provide evidence that the activity of these ACC synthases may not solely be controlled by the transcriptional activation of ACC synthase genes. PMID- 9207846 TI - Characterization of 26S proteasome alpha- and beta-type and ATPase subunits from spinach and their expression during early stages of seedling development. AB - Three kinds of cDNAs encoding 26S proteasome subunits have been cloned from spinach (Spinacia oleracea L.). These genes, designated as SOPSC8, SOPSC1 and SOPRS7, encode an alpha-type and a beta-type subunit of the 20S catalytic core, and an ATPase subunit of the 19/22S regulatory complex, respectively. The deduced protein sequences showed high sequence similarities to other proteasome alpha- and beta-type and ATPase subunit proteins. Southern blot analysis indicates that there are additional members of these dispersed proteasome families in the spinach genome. These three subunit genes are expressed simultaneously during germination and reach a maximum one day after sowing followed by a decline. The expression of these genes also increases during cotyledon senescence. PMID- 9207845 TI - The ribosomal protein P0 of soybean (Glycine max L. Merr.) has antigenic cross reactivity to soybean seed lectin. AB - Soybean (Glycine max L. Merr.) mutants lacking the ability to produce the lectin normally found in soybean seeds (SBL) are designated Le-. A protein of higher molecular weight that cross-reacts with antibodies raised to SBL was found at nearly equivalent levels in roots, hypocotyls, and leaves, and at lower levels in cotyledons and dry seeds of both Le+ and Le- soybean cultivars. Earlier work suggested that this protein was a novel lectin. Clones isolated from a Le- soybean root cDNA library produced a cross-reacting protein of the same size in Escherichia coli. Sequence analysis of these clones revealed a high degree of similarity to the ribosomal protein P0. The cross-reacting protein co-purified with ribosomes, and a monoclonal antibody raised to purified brine shrimp P0 cross-reacted to the same protein. The protein showed no lectin activity in a hemagglutination assay, nor did it bind to an N-acetyl-D-galactosamine affinity column. On the basis of this evidence, we conclude that the SBL-cross-reacting protein is not a lectin but a homologue of the ribosomal protein P0. Consequently, Le- soybeans must produce a lectin that is dissimilar to SBL at both the DNA and amino acid levels and we suggest that it is this lectin which is involved in nodulation. PMID- 9207847 TI - Potato cysteine proteinase inhibitor gene family: molecular cloning, characterisation and immunocytochemical localisation studies. AB - Potato cysteine proteinase inhibitors (PCPIs) represent a distinct group of proteins as they show no homology to any other known cysteine proteinase inhibitor superfamilies, but they all belong to the Kunitz-type soybean trypsin inhibitor family. cDNA clones for five PCPIs have been isolated and sequenced. Amino acid substitutions occurring in the limited regions forming loops on the surface of these proteins suggest a further classification of PCPIs into three subgroups. Accumulation of PCPI was observed in vacuoles of stems after treatment with jasmonic acid (JA) using immunocytochemical localisation, implying that these inhibitors are part of a potato defence mechanism against insects and pathogens. Genomic DNA analysis show that PCPIs form a multigene family and suggest that their genes do not possess any introns. PMID- 9207848 TI - Cloning and characterisation of a carrot cDNA coding for a WD repeat protein homologous to Drosophila fizzy, human p55CDC and yeast CDC20 proteins. AB - The present study describes the isolation of a cDNA coding for a carrot protein of 450 amino acids that contains WD repeats (DcWD1) and is homologous to Drosophila melanogaster fizzy protein, mammalian p55CDC and yeast Cdc20p. As for the known related proteins, sequence conservation concerned the majority of the polypeptide except the far N-terminus. Results of Southern blot analysis with genomic DNA under high stringency conditions showed the occurrence of a single gene. Northern blot analyses revealed the accumulation of DcWD1 mRNA in all tested tissues (leaves, petioles and hypocotyls, apical meristems, roots and suspension cultured cells), though at a different extent. Lack of induction of relevant transcripts in proliferating auxin-stimulated hypocotyls suggests a mode of expression not strictly related to the cell proliferation. PMID- 9207849 TI - Characterization of a barley gene coding for an alpha-amylase inhibitor subunit (CMd protein) and analysis of its promoter in transgenic tobacco plants and in maize kernels by microprojectile bombardment. AB - A gene coding for a barley CMd protein was isolated from a genomic library using a cDNA probe encoding the wheat CM3 protein. Promoter sequence analysis reveals motifs found in genes specifically expressed in endosperm and aleurone cells, as well as TATA and other putative functional boxes. 720 bp of the Hv85.1 CMd protein gene promoter, when fused to a gus coding region, were unable to direct GUS activity in the seeds of transgenic tobacco plants. In contrast, the same construction delivered into immature maize kernels by microprojectile bombardment was able to direct expression of GUS in the outermost cell layers of maize endosperm in both a tissue-specific and a developmentally determined manner. PMID- 9207850 TI - DNA binding and bending by a chloroplast-encoded HU-like protein overexpressed in Escherichia coli. AB - The Guillardia theta chloroplast hlpA gene encodes a protein resembling bacterial histone-like protein HU. This gene was cloned and overexpressed in Escherichia coli cells, and the resulting protein product, HlpA, was purified and characterized in vitro. In addition to exhibiting a general DNA-binding activity, the chloroplast HlpA protein also strongly facilitated cyclization of a short DNA fragment in the presence of T4 DNA ligase, indicating its ability to mediate very tight DNA curvatures. PMID- 9207852 TI - A chloroplast derived trnH gene is expressed in the mitochondrial genome of gramineous plants. AB - We reported previously that the mitochondrial sequence that contains the chloroplast-derived trnH gene has been highly conserved in the region around one terminus of the junction between chloroplast-derived and mitochondrion-specific sequences in most of the gramineous plants analyzed [15]. The results of RT-PCR, northern hybridization, in vitro capping and ribonuclease protection experiments show that the chloroplast-derived trnH gene is transcribed from a putative promoter that is located in the mitochondrion-specific sequence. Gene expression in this region seems to be correlated with the conservation of the sequence at the junction between the chloroplast-derived fragment and the mitochondrion specific sequence. PMID- 9207851 TI - Characterisation of a pea hsp70 gene which is both developmentally and stress regulated. AB - A pea pod cDNA library was screened for sequences specific to lignifying tissue. A cDNA clone (pLP19) encoding the C-terminal region of a hsp70 heat shock protein hybridised only to pod mRNA from pea lines where pod lignification occurred. Expression of pLP19 was induced by heat shock in leaves, stems and roots of pea and chickpea plants. Four different poly(A) addition sites were observed in cDNAs derived from the same gene as pLP19. This gene was fully sequenced; unlike most hsp70 genes, it contains no introns. The 5'-flanking sequence contains heat shock elements and other potential regulatory sequences. PMID- 9207853 TI - Expression of human muscarinic cholinergic receptors in tobacco. AB - We expressed human m1, m2 and chimeric muscarinic cholinergic receptors (MAChR) in tobacco plants and in cultured BY2 tobacco cells using Agrobacterium-mediated transformation. The membranes of most transgenic plants and calli bound muscarinic ligands with appropriate affinities, kinetics and pharmacologic specificity, as determined by direct and competitive binding measurements using the muscarinic ligand [3H]quinuclidinyl benzylate (QNB). Membranes of untransformed plants and calli or those transformed with vector alone did not bind [3H]QNB. Preliminary experiments did not suggest regulation of endogenous plant G protein signalling pathways by the recombinant receptors. Membranes from one callus clone expressed m1 MAChR at the level of 2.0-2.5 pmol [3H]QNB bound per mg membrane protein, more than the number of m1 MAChR in mammalian brain and comparable to that expressed in Sf9 insect cells using baculovirus vectors. This work demonstrates high level expression of active G protein-coupled receptors in plants, such that signaling might be genetically reconstituted by co-expression of appropriate G proteins and effectors. PMID- 9207854 TI - Expression of C3 and C4 photosynthetic characteristics in the amphibious plant Eleocharis vivipara: structure and analysis of the expression of isogenes for pyruvate, orthophosphate dikinase. AB - Eleocharis vivipara, a unique leafless amphibious sedge, adopts the C4 mode of photosynthesis under terrestrial conditions and the C3 mode under submerged aquatic conditions. To analyze the molecular basis of these responses to the contrasting environments, we isolated and characterized two full-length cDNAs for a key C4 enzyme, pyruvate, orthophosphate dikinase (PPDK; EC 2.7.9.1). The isogenes for PPDK, designated ppdk1 and ppdk2, were highly homologous to one another but not identical. The PPDK1 protein, deduced from the nucleotide sequence of the cDNA, contained an extra domain at the amino terminus which, presumably, serves as a chloroplast transit peptide, while PPDK2 lacked this extra domain. It seems likely, therefore, that the ppdk1 and ppdk2 genes encode a chloroplastic and a cytosolic PPDK, respectively. Genomic Southern blot analysis revealed the existence of a small family of genes for PPDK in the genome of E. vivipara. Northern blot analysis indicate that both chloroplastic and cytosolic genes for PPDK are expressed simultaneously in the culms, a photosynthetic organ, of E. vivipara and that the pattern of expression of these genes differs between the growth forms. PMID- 9207855 TI - Activins, inhibins, and follistatins: further thoughts on a growing family of regulators. AB - Inhibin, a feedback inhibitor of pituitary FSH secretion, and its homodimer, activin, have been the subject of a growing body of literature in the last 5 years. These factors play a role not only in endocrine feedback in the reproductive system but also in paracrine and autocrine regulation of both reproductive and nonreproductive organs, including the liver, kidney, and brain. Additionally, the messages coding for both subunits and their receptors are exquisitely regulated, both spatially and temporally, during embryonic development. The cloning of a family of activin receptors; the development of specific immunoassays for inhibins A and B, and activins A and B; the description of alpha subunit, beta subunit, and receptor loss of function transgenic mouse models; and the cloning of two new beta subunit homologs have increased our understanding of the possible roles this complex family of proteins plays in development and endocrine function. This review largely confines itself to the roles of inhibins and activins in the male and female reproductive system, and is intended as an update to a 1992 review published in this journal. PMID- 9207856 TI - The vitamin D receptor--structure and transcriptional activation. AB - The vitamin D receptor (VDR) is an integral part of the body's calcium regulatory system. In this review, recent advances in the understanding of VDR structure and function are discussed. Both direct mutagenesis studies on the VDR and structural studies of related receptors have been reviewed. Recent insights into DNA binding and ligand binding by the VDR are discussed, along with implications for gene regulation by the VDR. The potential role of co-activators and co-repressors in VDR-regulated transcription are discussed, and avenues of future research proposed. PMID- 9207857 TI - A reappraisal of the role of zinc in life and death decisions of cells. AB - There is a great deal of interest in chemicals and biochemicals that can modulate apoptosis. As will be discussed, zinc, an essential trace element, can induce as well as block apoptosis. High concentrations of extracellular zinc (500-1000 microM) have frequently been used to block apoptosis or programmed cell death in a variety of systems. Early investigators provided evidence that this concentration of zinc could block DNA fragmentation that is often associated with apoptosis. Since zinc plays a role in many aspects of cell function, there are probably many sites in a death pathway that zinc could potentially modulate. In the case of glucocorticoid-mediated apoptotic death, new evidence presented herein indicates that high zinc can also block the binding of steroids to the glucocorticoid receptor thereby inhibiting the death signal itself. In this case, zinc probably binds to the vicinal cysteines in the receptor ligand binding site thereby blocking binding of glucocorticoid. Indeed, glucocorticoid-induced apoptosis in thymocytes has become one of the most frequently studied systems and is a focal point of this review. Studies herein will show that unlike zinc other trace-like metals such as nickel, copper, cadmium, and gold do not afford thymocytes protection against the DNA fragmentation induced by glucocorticoid mediated cell death. Interestingly, in attempting to determine if lower or more physiological concentrations of zinc could provide protection against apoptosis, it was found that 80-200 microM zinc could actually induce death in 40% of CD4+ CD8+ alpha beta TCR10CD3(10) thymocytes. From these experiments one might have been optimistic that zinc could, indeed, be a modulator of cell death. However, this thought has been overshadowed by growing evidence that zinc does not provide long-term protection to so-called surviving cells. PMID- 9207858 TI - Increased longevity, reduced fecundity, and delayed development in fruitfly (Zaprionus paravittiger) fed on butylated hydroxy anisole. AB - Oxygen free radicals are generated as a by-product during normal metabolism, and they cause damage to proteins, lipids, and DNA in organisms. The defense system of the body counteracts these highly reactive chemical moieties and neutralizes them. However, a small fraction of free radicals escapes, which causes lipid peroxidation and hence aging of the organism. It has been hypothesized that, if the free radicals are arrested/reduced, then aging can be delayed or life span could be enhanced. To test the above hypothesis, we fed butylated hydroxy anisole (BHA) to a drosophilid insect, Zaprionus paravittiger, and observed its effect on life span, fecundity, and developmental period. The insects were reared and maintained on standard corn meal agar (CMA) medium at 26 degrees +/- 2 degrees C. Various concentrations (1, 5, 10, 25, 50, and 100 mM) of BHA were mixed with CMA medium, and the cultures were reared and maintained on these mixtures to study the life span of insects. Survivor curves showed that lower concentrations (5, 10, 25 mM) of BHA increased the life span, while higher concentrations (50, 100 mM), which were rather toxic, decreased life span. The most suitable concentration was 10 mM, which increased median (LT50) (27% and 15% in male and female) and maximum (LT100) (18% and 27% in male and female) life spans of insects maximally. Females exhibited longer life spans compared with males. The cultures were fed on the optimal concentration (10 mM) of BHA to study its effect on developmental period and egg-laying rate. The total developmental period was delayed by 7.2%, and egg-laying rate was reduced by 19.7% on BHA feeding. The extension in developmental period and reduction in egg-laying capacity could be the contributory factors to the favorable effect of BHA on life span. PMID- 9207859 TI - Human placenta does not Reduce AZT (zidovudine) to 3'-amino-3'-deoxythymidine. AB - Human placental tissue and human trophoblast cells (JAr) were examined after exposure to the anti-HIV nucleoside analog AZT (Zidovudine) for the presence of 3'-amino-3'-deoxythymidine (AMT), a toxic catabolite. Placental cells were exposed to 7.6 mM AZT for 48 hr, and placental lobular tissue was perfused with 3.8 mM AZT for 14 hr. Cell homogenates were prepared, and supernatants were subjected to HPLC analysis. Despite large cellular concentrations of AZT, AMT was not detected in any of the samples analyzed. Exposure of JAr cells to this concentration of AZT produces a 72% inhibition of cell proliferation when compared with unexposed controls. Based upon the results of the current study, AMT was not formed by placental cells exposed to AZT and, thus, not a mechanism for toxicity after in vitro exposure to AZT. PMID- 9207860 TI - Ascorbyl-2-sulfate compared with ascorbic acid in Atlantic salmon: uptake and distribution confirmed by mass spectroscopy. AB - This paper reports an in-depth approach that identifies ascorbyl-2-sulfate (AS) in gastric, blood, liver, and muscle tissues of Atlantic salmon (Salmo salar). To insure the identify of the AS, the study utilized the latest high-performance liquid chromatography (HPLC) technology plus electron ionization mass spectrometry (EIMS). Just before saltwater adaptation stage, juvenile Atlantic salmon were force-fed AS, ascorbic acid (AA), and a molecular-equivalent combination of the two. After tissue analyses for AA and AS were performed by HPLC separation, the HPLC peaks were identified by EIMS. The data collected in this study indicate that Atlantic salmon can absorb AS through the gastric tissue when forced-fed AA and AS as described. The data also indicate that AS is transported through the blood to the liver. There is evidence to indicate that AS is converted to AA in the livers of these salmon. In addition, the muscle tissue contained a large portion of AA and AS. PMID- 9207861 TI - Modulation of the antibody response to sheep red blood cells in normal and immunodeficient XID mice by myo-inositol. AB - Over the dose ranges tested, dietary vitamin A and myo-inositol induce similar changes in murine fetal growth and development. Because vitamin A is also known to affect the immune response, studies were conducted to determine if dietary myo inositol might have an effect on antibody production. It was found that in vivo in inbred mice myo-inositol (4 mg/g of diet) accelerated the rate of appearance of plaqueforming cells (PFC) in a primary response to sheep red blood cells (SRBC). In vitro, myo-inositol accelerated the rate of appearance of colonies of anti-SRBC PFC (foci) and significantly increased the number of PFC per colony, but did not affect the number of foci per culture noted at the end of the culture period, myo-inositol had no effect on the PFC IgM:IgG ratio following a single exposure to the agent, but exposure to myo-inositol in vivo and in vitro resulted in a decrease in the number of IgM PFC per focus in a primary response and IgM and IgG PFC per focus in a secondary response. Based on studies suggesting that myo-inositol or a phosphorylated metabolite might act downstream from Bruton's tyrosine kinase (Btk) in a signal transduction pathway in B cells, immunodeficient CBA/CaHN-XID/J mice were fed a standard diet or the same diet supplemented with 0.4% myo-inositol. Mice given the supplemented diet produced significantly more IgM anti-SRBC antibody than did XID mice given the control diet (4.3 +/- 2.5 vs 1.7 +/- 2.8, 1/log2), and produced approximately the same amount as immunocompetent controls (2.9 +/- 0.9). When rechallenged with SRBC, XID mice given supplemental inositol produced significantly more IgM antibody than did the XID and immunocompetent controls (3.6 +/- 0.5 vs 1.8 +/- 1.1 and 1.5 +/- 0.7, respectively). Added dietary inositol did not have a significant effect on primary or secondary IgG responses to SRBC, which remained impaired. These results suggest that dietary myo-inositol or a derivative may be able to modulate B-cell IgM responses by interacting within the inositol second messenger system downstream from Bruton's tyrosine kinase. PMID- 9207862 TI - Dapsone decreases the cumulative incidence of diabetes in non-obese diabetic female mice. AB - Dapsone (4,4'-diaminodiphenyl sulfone) has a large clinical experience due to its antimicrobial effects against Mycobacterium leprae, the causative agent of leprosy, and is used clinically where inflammation mediated by neutrophils is perceived to play a role. We administered dapsone in two concentrations (0.001% and 0.0001% w/w of diet) to 30 female non-obese diabetic (NOD) mice to explore the effect of dapsone on the development of IDDM following either a 1-week pulse or 20 weeks of continuous oral dapsone administration. Those mice receiving either the high or low doses of dapsone in the continuous group had a significantly reduced cumulative percentage of onset of IDDM. One of the seven mice given 0.0001% dapsone became diabetic (age 25 weeks), while none of the eight high dose (0.001%) mice developed the disease. Histological examination of pancreatic sections revealed islet infiltration in all groups of animals. The pulse and continuous experiments showed no statistically significant difference in the frequency or severity of lymphocytic infiltration. Dapsone administration did not inhibit growth, and growth rates were greater in those animals receiving the higher dapsone dose compared with the lower dose comparable to controls. We studied whether dapsone influenced murine lymphocyte function in addition to the published effects of the drug on neutrophils. At doses approximating those achieved in vivo (0.4 and 2 micrograms/ml), dapsone was found to inhibit murine splenocyte IL-2 and IL-4 secretion in response to concanavalin A. In view of the wide clinical experience with dapsone, randomized trials of the drug in new onset diabetes may be warranted. PMID- 9207863 TI - Macrophage function in mice with a mutation at the microphthalmia (mi) locus. AB - Microphthalmic (mi/mi) mice are unpigmented, osteopetrotic, mast cell deficient, and exhibit diminished gastric acid secretion and natural killer cell activity. In order to assess the effect of this mutation on macrophage function, we studied superoxide (O2-) and nitric oxide (NO) production, superoxide dismutase (SOD) activity, phagocytic capacity, and tumor cell killing by peritoneal macrophages from mi/mi mice and normal (+/+) litter mates. Macrophages from mi/mi mice, upon activation with phorbol myristate acetate (PMA), generated significantly higher amounts of O2- than did macrophages from their +/+ litter mates. The phagocyte respiratory burst as assessed by nitroblue tetrazolium (NBT) reduction assay also displayed a 2-fold enhancement in mi/mi macrophages. The assay of SOD activity revealed significantly lower levels in mi/mi macrophages than in the wild type. Furthermore, in comparison with controls, macrophages from mi/mi mice demonstrated significantly higher levels of NO production and increased capacity to lyse tumor cells upon activation with lipopolysaccharide (LPS) or gamma interferon (IFN-gamma). The mi mutation, therefore, is associated with reduced macrophage SOD activity, increased O2- and NO production, and enhanced capacity for tumor cell killing. The molecular mechanisms for these changes have not been identified. PMID- 9207864 TI - Cellulose derivatives and intestinal absorption of water and electrolytes: potential role in oral rehydration solutions. AB - The physicochemical and structural characteristics of several types of carboxymethylcellulose (CMC) and of methylcellulose (MC) were examined in relation to their capacity to modify water and sodium absorption in oral rehydration solutions (ORS) at various concentrations, using a jejunal perfusion procedure in rats. Comparison of intrinsic low-viscosity CMC of various degrees of substitution (DS) revealed that net water absorption increased as the DS was augmented. A stimulatory effect on sodium absorption occurred only at a low (2.5 g/l) CMC concentration. With products of medium DS, stimulation of net water and sodium absorption was observed only with low-viscosity CMC at 2.5 g/l, but not at 5.0 g/l. In perfusions with CMC of medium and high DS there was a reduction of water and sodium absorption, ultimately resulting in net sodium secretion with 5.0 g/l high-DS CMC. MC perfused at 5.0 or 10.0 g/l reduced net water absorption and reversed sodium transport from absorptive to secretory status. These results show that while low-viscosity-grade, low-DS CMC in low concentrations may facilitate solute uptake and concurrent water absorption from ORS by the jejunum, high intrinsic viscosity and possible chemical interaction of solutes with the modified celluloses tend to block water uptake and produce fluid stasis and electrolyte secretion. Thus, the data suggest that only certain types of CMC may be proabsorptive when added to ORS, while high-viscosity and high-DS CMC or MC induce electrolyte malabsorption and eventual catharsis. PMID- 9207865 TI - Dietary regulation of the renal sodium-phosphate (Na+/Pi) transporter during early ontogeny in the rat. AB - Phosphates are necessary for proper skeletal growth and function, as well as for growth and development of cells. Phosphate repletion depends partly on the function of the renal sodium-phosphate (Na+/Pi) transport system that functions to recover filtered urinary phosphate. It has been suggested that in order to meet the higher phosphate requirement of the developing animal, the weanling rat would have a greater adaptive response to chronic phosphate deprivation than the adolescent rat. The current study sought to characterize the adaptive response to dietary phosphate deprivation in terms of Na+/Pi transporter activity, and mRNA and immunoreactive protein levels. Weanling and adolescent rats were pair fed either a low-phosphate diet (LPD) or a control-phosphate diet (CPD) for 1 week. Maximal rates of transport (Vmax) were not different in weanling or adolescent rats on CPD (weanling 2.13 +/- 0.29 nmol/mg protein/10 sec, and adolescent 1.41 +/- 0.036 nmol/mg protein/10 sec, n = 3). K(m) values were not different in either group on CPD (weanling 0.15 +/- 0.08 mM Pi, and adolescent 0.22 +/- 0.13 mM Pi). There were no difference in mRNA abundance (Na+/Pi transporter/1B15 = 0.194 +/- 0.12 for weanling and 0.230 +/- 0.03 for adolescents, n = 3) or immunoreactive protein levels (Na+/Pi transporter/beta-actin = 0.232 +/- 0.01 for weanlings and 0.300 +/- 0.05 for adolescents, n = 3) in the two groups when fed CPD. After chronic Pi deprivation, the weanling rat showed a greater adaptive response than the adolescent as measured by Vmax values (weanling LPD/CPD = 2.01, P < 0.01; adolescent LPD/CPD not different; n = 3), mRNA signal intensity (weanling LPD/CPD = 1.86, P < 0.05; adolescent LPD/CPD not different; n = 3), and protein signal intensity (weanling LPD/CPD = 3.63, P < 0.01, and adolescent LPD/CPD 1.91, P < 0.05; n = 3). K(m) values were not affected by LPD. Immunohistochemical analysis of kidney cortex showed greater apical staining in both groups on LPD, with the increase being noticeably greater in the weanlings. Furthermore, two-way analysis of variance demonstrates a significant adaptive response in the weanling period in regard to maximum transport capacity (Vmax) and immunoreactive protein (Western), suggesting a synergistic effect between the developmental stage and low-phosphate diet. Therefore, it appears that the adaptive response is greater in the more rapidly developing animal (the weanling), and these results suggest a compensatory mechanism to conserve phosphate during periods of rapid growth. PMID- 9207866 TI - [Oncogen N-myc expression and measurement of DNA ploidy in neuroblastoma: a double staining flow cytometric analysis]. AB - When N-myc copy number was assessed by molecular biology, it has been proven that an amplification of the oncogene was a bad prognosis in childhood neuroblastoma, and the same goes for DNA-diploidy. This study concerns the development of a biparametric flow cytometric analysis of 2 neuroblastoma cell lines (SK-N-SH and IGR-N-91), which exhibited respectively 1 and 60 copies of the N-myc oncogene. An indirect immunofluorescence technique allowed N-myc oncoprotein staining and an isotypic control was used to assess the threshold of specific fluorescence. Simultaneously, a double staining with propidium iodide gave the nuclear DNA content. For both types of cells, the level of N-myc expression was calculated as a fluorescence index (IF). IF for IGR-N-91 appeared 2.5 times higher than those of SK-N-SH. This fluorescence index increases significantly during the exponential growth of N-myc amplified cells, whereas it does not vary for SK-N SH. During IGR-N-91 tumoral evolution, the cell line which derived from murine heart metastasis was the only one to show an increased IF. When applied to 10 neuroblastoma cell suspensions, this double staining showed an high IF for only 1 N-myc amplified case. A poor cell yield after tumoral dissociation and too much debris did not allowed the calculation of IF for half of them, which hampered a routine development of this technique. PMID- 9207867 TI - [Oncogenic activation of p21ras or pp60c-src in human colonic Caco-2 cells induces post-translation alterations of syndecan-1]. AB - The protein encoded by ras and src protooncogenes are frequently activated in a constitutive state in human colorectal cancer. In this study, we investigated the effect of oncogenic p21ras and Py-MT/pp60c-src on the synthesis of syndecan-1, a membrane anchored proteoglycan playing a role in cell-matrix interaction and neoplastic growth control. To this end, we used Caco-2 cells transfected with an activated (Val-12) human Ha-ras gene or the polyoma middle T (Py-MT) oncogene, a constitutive activator of pp60c-src tyrosine kinase activity. As compared to control vector-transfected Caco-2 cells, both oncogene-transfected cells exhibited: (1) a decrease in syndecan-1 specific activity; (2) a decrease in size and sulfation of syndecan-1 ectodomain glycosaminoglycan side chains; and (3) an active heparanase specifically degrading the heparan sulfate chains. In conclusion, the tumorigenic progression induced by oncogenic p21ras or Py MT/pp60c-src is associated with marked alterations of syndecan-1 at the post translational level. PMID- 9207868 TI - [Adjuvant chemotherapy with mitoxantrone, cyclophosphamide and 5-fluorouracil in breast neoplasms: therapeutic life]. AB - The chemotherapy side-effects are insufficiently documented while they strongly condition patients' quality of life. The aim of the study was to assess by means of a self-administered questionnaire the somatic symptoms experienced by breast cancer patients during their NCF (mitoxantrone + cyclophosphamide + 5 fluorouracil) chemotherapy and to demonstrate the interest of this self-report by comparing the frequency of side-effects assessed by the patients to that noted by the physicians in medical records. The study was carried out among 44 patients receiving their chemotherapy + radiotherapy at the Paoli-Calmettes Institute (marseille) between July 1994 and May 1995. The questionnaire comprized of 17 symptoms evaluated in terms of frequency, duration/severity and distress. The most frequent symptoms are: hair loss and nausea (75%), hot flush (57%), lack of appetite and headache (46%) associated with distress in 67 to 100% of cases. Their frequency was underestimated by the physicians in medical records. This study showed a large discordance patient-physician in the assessment of chemotherapy side-effects. The type of tool presented in this study could complement the usual scales of toxicity that do not provide an estimation of true patients' experience. PMID- 9207869 TI - [Adjuvant combined radiochemotherapy: a feasibility study of a new strategy in stages I and II]. AB - Adjuvant radiotherapy is the rule after conservative surgery for breast cancer. Furthermore, an anthracycline-based chemotherapy is recommended in node-positive patients and in poor prognosis tumors. The optimal schedule of treatment has yet to be determined, but ideally, none of these therapeutic modalities should be delayed. We have therefore conducted a feasibility trial using post-operative concurrent chemoradiation therapy with an anthracenedione. Between May 1990 and October 1994, 154 patients with stage I or II breast cancer who had benefited of either limited or radical surgery were treated with adjuvant concurrent chemoradiotherapy. Radiotherapy consisted of 50 Gy in 25 fractions over 5 weeks to the chest wall or the breast, and to the supraclavicular and internal mammary lymph nodes. When indicated, a boost of 15 Gy was then delivered to the primary tumor bed (n = 75). Starting on the first week of radiotherapy, combined chemotherapy with 5-fluorouracil, mitoxantrone, and cyclophosphamide was administered at 21-day intervals, for 4 to 6 cycles. Compliance to therapy was excellent. Median radiotherapy dose was 49.5 Gy to the chest wall or breast, and to the lymph nodes, and 14.2 Gy to the tumor bed. Chemotherapy was given at full dose in over 80% of the cases and the 21-day interval between cycles was respected in 31%. In 45% of the cases, a 28-day interval was required due to toxicity, and at least one interval longer than 28 days was necessary in the remainder of the patients. Main toxicities were nausea and vomiting (20.8%) and grade 3-4 neutropenia (12.3%). Grade 1 cutaneous toxicity occurred in 62.3% of the cases, and severe grade 3 radiation dermatitis requiring temporary interruptions of therapy in 4.5%. With the exception of one case of grade 3 acute cardiac toxicity, there was no other severe side-effects. In conclusion, this pilot study demonstrates the feasibility of concurrent chemoradiation therapy with an anthracenedione for stage I and II breast cancer in the adjuvant setting. Whether this approach compares favorably with standard sequential therapy in terms of long-term results remains to be determined and should be assessed in a phase III trial. PMID- 9207870 TI - [Axillary lymphadenectomy prepared by fat aspiration versus functional axillary lymphadenectomy: preliminary results of a randomized prospective study]. AB - The objective of this study is to compare morbidity between 2 surgical procedures of axillary clearance: functional lymphadenectomy by classical dissection versus axillary dissection prepared by liposuction (Suzanne's procedure). Two hundred consecutive patients treated for breast cancer were included in a prospective randomized trial between 1st January, 1995 and 31st January, 1996 (Huriet's law). The assessment (number of nodes, postoperative stay, drainage duration, rate of seromas, number of complications, evaluation of mobility and sensitive disorders) was done on the first, fifth, tenth and thirty postoperative days. There is no significant difference between the 2 groups. The rate of seromas decreased significantly only for fat patients (8/25 versus 21/34, p < 0.05) and for the patients treated with radical mastectomy (17/37 versus 28/39, p < 0.05). In this preliminary study, liposuction does not change postoperative effects of axillary clearance, except for fat patients or after total mastectomy. The liposuction seems to facilitate a better anatomical dissection and a better preservation of the nervous and vascular elements. PMID- 9207871 TI - [Preventive analgesic effect of intraoperative administration of ibuprofen arginine on postmastectomy pain syndrome]. AB - The efficacy of preemptive analgesia on postoperative pain is discussed. From experimental neurophysiological data, the present policy of preventive analgesia aims at precluding modifications of the nervous system secondary to a nervous lesion and the appearance of chronic pain, particularly of the neurogenic kind. The post-mastectomy pain syndrome (PMPS) falls within the realm of neurogenic pain and is still poorly understood and underestimated. This study evaluated the preemptive effect of a perioperative administration of an oral non steroid anti inflammatory, the ibuprofen-arginine, on PMPS. Thirty patients scheduled for partial or total mastectomy with axillary dissection were prospectively and randomly assigned to 2 groups. The ibuprofen-arginine group (group I) (n = 15), received an oral administration of 400 mg of ibuprofen-arginine, 90 min before surgery, 2 h after surgery and then every 8 h in the first 32 postoperative hours. The control group (group C) received in doubled blind a placebo at the same time. At 6 months, we looked after pain or dysesthesia. We confirmed the diagnosis of PMPS in presence of association of diagnosis criterias. Fourteen patients in each group have been included. Eighty-six percent of the patients (13 patients in group I and 11 patients in group C) presented at 6 months dysesthesia of the upper member ipsilateral to the mastectomy and/or the operated breast, appearing either immediately or after a laps of time. Nine patients (group I) and 6 patients (group C) had PMPS. Postoperative radiotherapy and lymphoedema were statistically associated with PMPS (p = 0.019 and p = 0.011). The perioperative preventive administration of a non-steroid anti-inflammatory drug reduces neither the incidence of pain in the first post-operative months, nor the appearance of PMPS at 6 months. These results suggest that others factors than the nervous lesion may play a role in the occurrence of PMPS, as radiotherapy, lymphoedema, but also psychosocials factors. PMID- 9207872 TI - [High-dose chemotherapy in relapse of medulloblastoma in young children]. AB - Craniospinal irradiation is the gold standard treatment used in non metastatic medulloblastoma as prophylaxis against central nervous system (CNS) metastases. However, given the severe late effects caused by this procedure in children under 3 years of age, most pediatric oncologists are currently treating these patients with conventional chemotherapy in order to postpone or even avoid irradiation. In the French Society of Pediatric Oncology (SFOP) this attitude has been adopted since 1990. Among the patients treated without radiotherapy, 20 relapsed while on conventional chemotherapy and were entered in a study of high-dose chemotherapy (HDC) followed by autologous bone marrow transplantation (ABMT). Their median age at diagnosis was 23 months (range: 5-71 months) and the relapse occurred at a median time of 6.3 months after the initiation of chemotherapy. Complete surgical removal of the local relapse was the first treatment in 4/20 patients who were not evaluable for response. Sixteen of the 20 patients had measurable disease at the primary site (9 patients), or at metastatic sites (3 patients) or both (4 patients). The conditioning regimen consisted of combination busulfan 600 mg/m2 over 4 days and thiotepa 900 mg/m2 over 3 days. After recovery from aplasia, patients with a local relapse received local radiotherapy limited to posterior fossa. Among the 16 patients with measurable disease, 6 complete responses, 6 partial responses, 3 non response, were observed following HDC (response rate 75%). One patient was not evaluable. For the 20 patients, the event free survival (EFS) is 50%. Among the surviving patients, the median follow-up is 39.5 months post BMT (range: 21-92 months). Ten patients who developed a local relapse or local progression are alive with non evidence of disease (NED) without craniospinal irradiation. Among the 7 patients who developed a metastases or progression of metastases, only 1 is alive. Toxicity was high but manageable. One complication-related death occurred 1 month post BMT. With a 75% response rate, this HDC proved to be very efficient in relapsed medulloblastoma. A longer follow up is necessary to demonstrate whether, after a local relapse, HDC could replace craniospinal irradiation as prophylaxis against CNS metastases. PMID- 9207873 TI - [Evaluation of tumoral and lymph node response to neoadjuvant chemotherapy in undifferentiated nasopharyngeal carcinoma]. AB - Between January 1994 and June 1995, 19 cases of nasopharyngeal carcinoma, classified N2-N3 (AJC/UICC 1987) were treated by neoadjuvant chemotherapy including cisplatin 100 mg/m2 on day 1 and epirubicin 80 mg/m2 on day 1. Following course started on day 21. A clinical and scannographic evaluation was made after 3 courses of chemotherapy. Fifty-eight percent of the patients were N3. Seventy-four percent were T3-T4. Tolerance to chemotherapy was good in 100% of the cases. A functional improvement was obtained in 14 among 16 patients who were initially symptomatic. For lymph nodes, an objective response (OR) was observed in all patients. The response was complete (CR) in 53%. A regression of primary tumor was obtained in 68% of the patients, but it was complete in 16% only. Sixty-nine percent of the patients has have a tumoral OR have a lymph node CR. Ninety percent of the patients with a complete lymph node response have a tumoral regression more than 50%. A correlation seems likely between the importance of the tumoral and the lymph node responses. PMID- 9207874 TI - Lung resection for recurrence after pneumonectomy for metastases. AB - Resection of pulmonary recurrences after pneumonectomy for metastases is exceptional. Nevertheless in carefully selected patients surgery on the residual lung might be successfully performed. From January 1987 to February 1996, 5 patients underwent metastasectomy on single lung after pneumonectomy performed for the same metastatic disease. There were 3 male and 2 female with a mean age of 38 years at the time of surgery on single lung. All patients had a FEV1 > 40%. One patient (n degree 1) had 2 consecutive operations (wedge resections) on the right lower lobe followed 17 months later by right inferior lobectomy for metastases of soft tissue sarcoma. Three patients had only an operation on the residual lung (patient n degree 2 had 2 wedge resections for carcinoma; patient n degree 3 had 7 wedge resections for carcinoma; patient n degree 4 had 6 wedge resections for osteogenic sarcoma). The last patient (n degree 5) had 2 wedge resections on the right upper lobe and a large wedge resection on the right lower lobe for metastases of malignant corticosurrenaloma using a cardiopulmonary femoro-femoral by-pass without cardiac arrest. She postoperatively developed a right lower lobe venous infarction treated subsequently with a completion right lower lobectomy. She died in the postoperative course from cardiorespiratory insufficiency. The other patients had an uneventful postoperative course. Two patients (n degree 2 and n degree 4) died of their disease 14 and 12 months respectively after the surgery on the residual lung; by contrast 2 patients (40%) (n degree 1 and n degree 3) are still alive without recurrences 36 and 27 months after the last resection. In selected patients aggressive surgery for metastases on the residual lung can be successfully performed but the benefits in terms of long-term disease-free survival remain to be determined. PMID- 9207875 TI - [A phase II multicenter study of gemcitabine in non small cell lung cancers]. AB - Gemcitabine is a novel pyrimidine nucleoside whose activity has been demonstrated on solid tumors. We report here the results of a multicentre phase II trial of gemcitabine in chemonaive patients with inoperable non small cell lung cancer (NSCLC). Gemcitabine was given weekly at a dose of 1,250 mg/m2 administered as a 30 min intravenous infusion, for 3 weeks followed by 1 week of rest (1 cycle). All the 161 patients included were evaluable for toxicity and 151 of them were evaluable for efficacy. The majority of patients had a stage IIIb (31.1%) or stage IV (64.6%) disease; 10.6%, 83.2% and 6.2% of patients had a WHO performance status (PS) 0, 1, and 2, respectively. Adenocarcinoma accounted for 52.2% of cases and squamous cell carcinoma for 43.5% of cases. Three complete responses and 30 partial responses gave an objective response (OR) rate of 21.8% (95% confidence interval; 15.5-29.3%). All responses were validated by an independent Oncology Review Board. Median duration of response was 7.6 months. Median time to progression was 4.6 months (3.3 months in non responders and 7.6 months in responders). Median survival was 7.3 months in non responders and 13.4 months in responders (p < 0.001), which gave an overall median survival of 8.9 months (95% CI: 0.1-21.9 months) in the entire study population. An improvement of symptoms and personal state was also observed. Treatment was well tolerated. Neutropenia was the only dose limiting toxicity. WHO grade 3 or 4 neutropenia occurred in 19.6% and 5.7% of patients, respectively. With a 21.8% OR rate, this multicentre study confirms the activity of gemcitabine as a single agent in patients with inoperable NSCLC. Its good tolerance and original mode of action make gemcitabine a drug of choice in the therapeutic strategy of these tumors. PMID- 9207877 TI - [Clinical research and Huriet's law in medical oncology. Results of a French investigation among medical oncologists]. AB - In early 1996 a postal investigation was done among french medical oncologists to have their opinion about practical application of the law about clinical research. 58.3% answered, the majority of them having practice in public hospitals or cancer centers and teaching activities. All but 2 had been involved in such a research, half of them as main investigator of at least 1 trial. Two types of difficulties are declared: (1) methodological ones, to select patients, follow them up, fill in the different forms for administrative purpose; (2) relational ones, to inform patients, particularly on randomization of their treatment, due to potentially lethal course of their cancer. The majority of answering oncologists acknowledge the importance of such a research and the necessary rigorous methodology. Following their opinions, improvements should come from a better training of physicians and from an increase of human and material means to perform such a research. PMID- 9207876 TI - [Genetic alterations associated with pathologic differentiation of Wilms' tumors]. AB - Although several cytogenetic alterations have been associated with development of Wilms' tumor, a multigenic neoplasia, molecular mechanisms of its induction, development and maintenance remain to be elucided. In order to characterize these different steps we have developed a unique animal model of Wilms' tumor constituted by the MAV-1(N) induced avian nephroblastoma. This animal model led to the discovery in our laboratory of a new gene now (nephroblastoma overexpressed gene) which is overexpressed in all avian nephroblastoma. Expression of the human nov gene (novH), which is down-regulated by WT1, is also deregulated in Wilms' tumors. Nov characteristics suggest that it would play a role in the control of cellular proliferation and differentiation. Our observations also indicate that nov could be involved in the development of Wilms' tumors, and represent a marker of their differentiation state. PMID- 9207878 TI - [Clinical research and informed consent. Ethical and deontologic aspects]. PMID- 9207879 TI - [Primary mediastinal non-seminomatous germ-cell tumors: from clinics to biology]. AB - Primary mediastinal non-seminomatous germ-cell tumors (PMNSGCTs) are rare neoplasms that occur in young male adults. Incidence is evaluated about half that of extra-gonadal GCT. Their treatment is generally based on protocols used for testicular cancer, but with poorer results. Based on our experience of 40 patients with PMNSGCTs and data from the literature, we review here the clinical and biological data of these neoplasms. PMNSGCTs seem to constitute a specific entity, distinct from other GCT by the following criteria: true extra-gonadal origin, high incidence in patients with the Klinefelter's syndrome, over representation of the yolk-sac component, poorer chemosensitivity and survival compared to other GCT, frequent occurrence of non-treatment related hematological neoplasia. The finding of an isochromosome of the short arm of the chromosome 12 in the leukemic karyotype is one of the strongest argument for a common origin in the yolk-sac component of the PMNSGCTs and their associated leukemia. Treatment of PMNSGCTs is still a challenge and should be conducted by a well-trained medical team. PMID- 9207880 TI - [Verrucous carcinoma of the larynx. Diagnostic and therapeutic problems]. PMID- 9207881 TI - [Tuberculous meningitis: clinical, biological and x-ray computed tomographic comparison between patients with or without HIV infection]. AB - OBJECTIVES: Determine possible differences in clinical manifestations, laboratory findings and neuroimaging results in tuberculous meningitis patients with and without HIV infection. PATIENTS AND METHODS: We retrospectively reviewed data of 38 patients with positive cerebrospinal fluid cultures for Mycobacterium tuberculosis who were hospitalized in 3 university hospitals in Paris over the last 11 years. RESULTS: There were 24 HIV-infected patients and 14 without HIV infection. Mean CD4 lymphocyte count was 103 +/- 180/mm3 in the HIV group. Age (median age = 33 years for the HIV group vs. 53 for the non-HIV group), sex ratio (3 vs. 0.75), and prior history of tuberculosis (46% vs. 43%) were similar in both groups. Clinical presentation was similar for headache (83% in HIV group vs. 50% in non-HIV group; p = 0.02) and confusion (54% vs. 93% in non-HIV group p = 0.05). Serum natremia (mmol/l) (131 +/- 5 vs. 125 +/- 8; p = 0.024), white blood cell count (x 10(9)/l) (5.8 +/- 4.7 vs. 10.7 +/- 1.7; p = 0.37) and erythrocyte sedementation rate (mm/h) (68 +/- 34 vs. 31 +/- 35; p = 0.003) were significantly different in the 2 groups. Median cerebrospinal fluid findings were similar in the 2 groups: leukocytes (x 10(6)/l) (375 +/- 860 vs 218 +/- 250), glucose (mmol/l) (2.3 +/- 0.9 vs 2.7 +/- 1.9) and protein (g/l) (3.8 +/- 7.1 vs. 2.6 +/- 1.6). CT-scans of the brain were similar in the 2 groups. Mortality during hospitalization was similar (42% vs 36%; NS). CONCLUSION: HIV infection appears to have little impact on the presentation of tuberculous meningitis. PMID- 9207883 TI - [Factitious hyperchloremia disclosing bromide poisoning. 4 cases]. AB - BACKGROUND: When routine blood chemistry shows elevated chloride alone, another anion (bromide, iodine, fluoride) may be involved. CASE REPORTS: Hyperchloremia and decreased anion gap was observed in four patients. Chloremia ranged from 134 to 174 mmol/l at initial blood tests. Careful history taking led to the discovery of long-term use of calcium bromogalacto-glucomate. Specific assay with inductively coupled plasma mass spectrometry (ICPMS) confirmed the presence of bromide in the blood. Chloridemia returned to normal levels after discontinuing use of bromine. DISCUSSION: Bromism is not a common diagnosis. Risks include neurological and psychiatric disorders due to bromide diffusion through the blood brain barrier. Clinical manifestations have been described including skin lesions, digestive intolerance, and fever. Bromide is contained in certain prescription drugs. Patients should be warned against the adverse effects of overuse. PMID- 9207882 TI - [Topographical diagnosis of insulinoma of the pancreas. Value of the test of intra-arterial calcium infusion]. AB - OBJECTIVE: The localization of insulinomas is a central problem, because none of the imaging techniques has proved sufficiently reliable for the diagnosis of tumors smaller than 2 cms. PATIENTS AND METHODS: Calcium stimulation test was performed during selective pancreatic angiogram with calcium gluconate injection in the pancreatic arteries, e.g. gastroduodenal, superior mesenteric and splenic. A simultaneous catheterization of hepatic veins through the femoral vein allowed measurement of insulin level increment after calcium stimulation. RESULTS: In our experience, this test allowed a positive location of an insulinoma in three consecutive patients presenting with organic hypoglycemia, and was useful for surgeons while non-invasive imaging techniques failed to locate the tumor in any of the three patients. DISCUSSION: The calcium stimulation test is a reliable preoperative procedure, particularly when other imaging techniques fail to locate the tumor. However it remains invasive, and our data are still too preliminary to clearly define its place within imaging techniques. PMID- 9207884 TI - [Stevens-Johnson syndrome caused by doxycycline]. PMID- 9207885 TI - [Hypoglycemia during a treatment with interferon-alpha]. PMID- 9207886 TI - [Diaphragm paralysis and facial diplegia with albumin-cell count dissociation in acute ethylene glycol poisoning]. PMID- 9207887 TI - [Clinical value of interferon-alpha in the treatment of malignant hematologic diseases]. AB - The antiviral, antiproliferative and immunomodulating effect of interferon alpha (INF alpha) has led to its widespread use in malignant diseases. In hematology, the clinical effect of INF alpha has been proven empirically for several lymphoid malignancies--hairy cell leukemia, chronic lymphoid leukemia, multiple myeloma, follicular lymphoma--and also for chronic myeloproliferative diseases, particularly chronic myeloid leukemia. However, after 10 years of use, the impact of INF alpha on patient management compared with conventional treatments remains a matter of debate. Interest in cost-containment and the frequency of adverse effects after long-term treatment also raises many questions. A critical analysis of the role of INF alpha in each specific indication is thus required. PMID- 9207888 TI - [Attitudes and experience of French general practitioners towards HIV infection. National survey in 1996]. PMID- 9207889 TI - [Which cross-match before organ transplantation in 1997?]. AB - GOALS: The objective of pre-transplantation cross-matching is to detect in the recipient's serum solely those anti-donor antibodies which could have a deleterious effect on the grafted organ. It is important to avoid refusing organs on the basis of recipient antibodies which do not imply a risk of rejection. SEVERAL METHODS: In most laboratories cross-matching or compatibility tests made before organ transplantation are based on a complement-dependent microlymphocytotoxicity technique, sometimes sensitized with anti-human globulin serum, reactive against target T-cells. A positive results is reported if the reactions are week, but other methods are available. CHOICE OF SERA FOR CMX: This is essential. Several sera must be used: the most positive sera, the last serum and the current sera if the patient has been transfused between the date of the last serum harvested and the CMX. IMMUNOLOGICAL STATUS: There is wide agreement on the requirement for quality surveillance of the recipient's immunological status. This policy is the only way to effectively select sera to cross-match before transplantation, whatever the technique used, and thus improve transplantation outcome and reduce the number of rejections. CASE BY CASE: These prerequisites hold for all organs, but especially so for renal (and/or pancreas) grafts. For heart or heart-lung transplantations, emergency procedures may be needed. PMID- 9207890 TI - [Autoimmune myasthenia: recent physiopathological data]. AB - AUTOIMMUNE DISEASE: Myasthenia gravis is a one of the rare autoimmune diseases with a well-defined antigen. Autoantibodies directed against the muscular acetylcholine receptor (AChR) play a key role, blocking the acetylchoroquine binding site and provoking accelerated receptor degradation; complement destroys the post-synaptic membrane. Disease severity is correlated with extent of AChR loss. THYMUS: Involvement of the thymus is suggested by the beneficial effect of thymectomy and by histological and functional anomalies (modified cellular composition, abnormally activated lymphocytes, sensitivization of certain lymphocytes to AChR). "ANTIBODY NEGATIVE" DISEASE: The lack of anti-AChR antibodies detectable at immunoprecipitation, evidences the complexity of myastenia gravis pathogenesis. These "antibody-negative" forms are distinct from seropositive forms which have a high frequency of pure occular involvement and from infantile forms where the thymus regresses. "Antibody-negative" myastenia gravis might be mediated by antibodies directed against other endplate tardets. CLINICAL DIVERSITY: Our understanding of mechanisms responsible for the diversity of the clinical expression highlight the role of the AChR polymophism (with and adult and fetal expression). GENETIC PREDISPOSITION: Several factors involved in disease pathogenesis have been identified, including genetic factors: the HLA system, AChR alpha chain gene polymorphism, and immunoglobulin and T-cell antigen receptor gene polymorphism. PMID- 9207891 TI - [Treatment of malignant adrenal cortex carcinoma]. AB - UNCOMMON MALIGNANCY: Adrenocortical carcinoma is a very rare malignancy with poor prognosis. Median survival ranges from 12 to 25 months. Most clinicians recommend aggressive surgical management of either local or recurrent and metastatic disease. ANTICORTISOL AGENTS: Mitotane, the most tested agent against inoperable and metastatic adrenocortical carcinoma, procures overall response rates of 20 to 25%, but recent data do not support its use in the adjuvant setting. CHEMOTHERAPY: The efficacy of cytotoxic chemotherapy is low but new agents or associations (with platinum salts), new concepts (dose-intensification, MDR (the chemoresistance gene) reversing agents) may be useful and five some hope in this difficult disease. RADIOTHERAPY: The role of radiation therapy must be developed. PMID- 9207892 TI - Embryotoxicity/teratogenicity study with isomaltulose (Palatinose) in rats. AB - The embryotoxicity/teratogenicity of the natural sweetener isomaltulose (Palatinose) was studied in Wistar rats. Groups of 24 mated females were fed diets containing isomaltulose at levels of 0, 2.5, 5 and 10% from day 0 to day 21 of pregnancy. The dams were killed on day 21 of pregnancy. No maternal toxicity occurred and no effects on reproductive performance, nor on embryonic or foetal development, including visceral and skeletal examination, were seen in any of the groups fed isomaltulose. The dietary level of 10% isomaltulose was equivalent to about 7 g/kg body weight/day. PMID- 9207893 TI - Safety evaluation of lipase derived from Rhizopus oryzae: summary of toxicological data. AB - A lipase enzyme, obtained from Rhizopus oryzae produced by a fermentation process was subjected to a series of toxicological tests to document the safety for use as a food additive. The enzyme product was examined for acute, subacute and subchronic oral toxicity, and mutagenic potential. An extensive literature search on the production organism has also been conducted. No evidence of (sub)acute oral toxicity or mutagenic potential was found. Administration of the lipase at dosages of 50, 200 and 1000 mg/kg body weight/day for 90 days did not induce noticeable signs of toxicity. A few minor changes in the chemical composition of the blood in the highest dose group were of no toxicological significance. The no observed-adverse-effect level of the tox-batch in the subchronic toxicity study was 1000 mg/kg body weight/day. It can be concluded that no safety concerns were identified in the studies conducted with this lipase preparation derived from R. oryzae and produced under controlled fermentation conditions. PMID- 9207894 TI - Safety studies of a novel starch, pullulan: chronic toxicity in rats and bacterial mutagenicity. AB - This study was designed to assess the potential toxicity of pullulan, a starch like substance produced by Aureobasidium pullulans that is proposed for use as a texturizer for meat and meat substitutes and as a flavour substrate. Sprague Dawley rats (15/sex/group) were administered pullulan as a dietary admixture at levels of 1, 5 and 10% for a period of 62 wk. Control animals (15/sex) received untreated standard laboratory diet. The feeding study, originally intended to continue for 24 months, was terminated at 62 wk due to poor survival resulting from intercurrent pneumonia which was confirmed by histological findings. At 62 wk of treatment, all survivors were killed, complete gross post-mortem examinations were conducted on all animals, selected organs were weighed and organ/body weight and organ/brain weight ratios calculated. Mean body weights of all treated groups were comparable to controls. There were no indications of an adverse effect of pullulan on food consumption or food efficiency. At termination of the study, haematology and clinical chemistry values of treated animals were comparable to control values. There was no indication of pullulan-related toxicity in terminal organ and body weights. Macroscopic and microscopic examinations revealed no toxic lesions due to treatment. The mutagenicity of pullulan was assessed with and without metabolic activation in Salmonella typhimurium (TA100, TA1535, TA98 and TA1537). Pullulan did not increase the number of revertants per plate in any strain at any dose, including 10,000 micrograms/plate with or without metabolic activation, suggesting that it lacks mutagenic/carcinogenic potential on the basis of these results, it is concluded that pullulan lacks significant toxicological activity. The no-observed-adverse effect level was 10% (equal to or greater than 4450 mg/kg body weight/day in males and 5080 mg/kg body weight/day in females) which would support use in various foods as a substrate for flavours. PMID- 9207895 TI - Carcinogenicity study of beta-cyclodextrin in F344 rats. AB - The carcinogenicity of beta-cyclodextrin, a cyclic, water-soluble carbohydrate comprising seven glucose units, was examined in Fischer 344 (F344) rats. Groups of 50 males and 50 females were given the compound in their diet at concentrations of 0 (control), 2.5 or 5% for 104 wk. Surviving rats were then given a basal diet for a further 5 wk and killed at 109 wk. The dose levels were selected from the results of a 13-wk subchronic toxicity study. Dose-dependent inhibitory effects of beta-cyclodextrin on growth were observed in both sexes of the treated groups. The survival rates, mean survival times and range, however, demonstrated no significant differences between the control and treated groups. A variety of tumours developed in all groups, including the control group, but all the neoplastic lesions were histologically similar to those known to occur spontaneously in this strain of rat, and no statistically significant increase in the incidence of any tumour was found for either sex of the treated groups. Thus, it is concluded that under the present experimental conditions, the high dose, about 340-400 times higher than the current daily human intake from ingestion as a food additive and from pharmaceutical use, does not have any carcinogenic potential in F344 rats. PMID- 9207896 TI - Acute and subacute toxicity of tyramine, spermidine, spermine, putrescine and cadaverine in rats. AB - The acute and subacute toxicity of five biogenic amines-tyramine, spermidine, spermine, putrescine and cadaverine-were examined in Wistar rats. Tyramine and cadaverine had a low acute oral toxicity of more than 2000 mg/kg body weight. Putrescine had an acute oral toxicity of 2000 mg/kg body weight and spermidine and spermine each of 600 mg/kg body weight. All amines investigated caused a dose related decrease in blood pressure after intravenous administration, except for tyramine, where an increase was found. In 6-wk studies the biogenic amines were administered in the diet to groups of 10 male and 10 female rats. Tyramine and cadaverine were given at levels of 0, 200, 2000 or 10,000 ppm, spermine and putrescine at levels of 0, 200, 2000 or 5000 ppm and spermidine at levels of 0, 20, 200 or 500/1000 ppm in the first study and at levels of 0 or 10,000 ppm in a second study. Spermine was the most toxic. The high dose level showed a great number of changes, such as emaciation, aggressiveness, convulsions and paralysis of the hind legs. Growth, food intake and water intake were considerably decreased. Slight anaemia (males) and changes in plasma clinical chemistry occurred. The relative weights of the thyroid, adrenals, spleen and heart were increased and that of the liver decreased. Impaired kidney function, together with renal histopathological changes and changes in plasma electrolytes and urea, occurred with spermine. Histopathological examinations also revealed decreased glycogen content in the liver, reduction of spermatogenesis, severe depletion of splenic white pulp, acute involution of the thymus and moderate myocardial degeneration in the heart. Myocardial degeneration was also seen in one mid-dose male. Adverse effects were also observed in the top dose groups of all other amines. Decreased body weights associated with diminished food intake were generally seen. Slight increases in packed cell volume, haemoglobin concentration and thrombocytes occurred with cadaverine. With spermidine, decreased plasma creatinine, calcium and inorganic phosphate were observed and decreased potassium levels with cadaverine. The no-observed-adverse-effect level was 2000 ppm (180 mg/kg body weight/day) for tyramine, cadaverine and putrescine, 1000 ppm (83 mg/kg body weight/day) for spermidine and 200 ppm (19 mg/kg body weight/day) for spermine. PMID- 9207897 TI - Nitrite-induced adrenal effects in rats and the consequences for the no-observed effect level. AB - In a previous subchronic oral toxicity study with potassium nitrite, hypertrophy of the adrenal zona glomerulosa was observed for all nitrite levels examined including the lowest level of 100 mg/litre. This present study was carried out, therefore, to establish a no-observed-effect level (NOEL) for nitrite. Groups of 10 male and 10 female 6-wk-old Wistar rats received KNO2 at levels of 12.5, 25, 50, 100 or 3000 mg/litre or NaNO2 at levels of 81 or 2432 mg/litre in the drinking water for 13 wk. The nitrite content of the drinking water in the latter two groups was equal to that of the 100 and 3000 mg KNO2/litre groups, respectively. Potassium and sodium concentrations were equalized in the corresponding test groups with KCl and NaCl, respectively. General health, behaviour and survival were not affected by the ingestion of nitrite. Body weight and food and liquid intake were slightly decreased in the 3000 mg KNO2/litre and 2432 mg NaNo2/litre groups for both sexes. Methaemoglobin concentration was significantly elevated in rats of both high-dose nitrite groups in wk 4 and 12, while slight increases in a number of red blood cell variables occurred with 3000 mg KNO2/litre in females in wk 12. Relative kidney weights were increased in both high-dose nitrite groups. In wk 4, plasma aldosterone and corticosterone levels were slightly decreased in males with 2432 mg NaNO2/litre and plasma corticosterone in females with 3000 mg KNO2/litre but not in wk 13. Systolic blood pressure was not affected by nitrite. Microscopic examination revealed slight hypertrophy of the adrenal zona glomerulosa in animals of the 100 and 3000 mg KNO2/litre and of the 81 and 2432 mg NaNO2/litre groups, the incidence and degree being dose related. The results obtained with 100 and 3000 mg KNO2/litre in the drinking water were comparable with those found at the same levels in the previous 90-day study. The effects with sodium nitrite were similar to those observed with potassium nitrite. The biological significance of the adrenal zona glomerulosa hypertrophy is discussed. It is concluded that the NOEL of KNO2 is 50 mg/litre in the drinking water, equivalent to about 5 mg/kg body weight/day. PMID- 9207898 TI - Inhibition of the mutagenicity of 2-nitrofluorene, 3-nitrofluoranthene and 1 nitropyrene by flavonoids, coumarins, quinones and other phenolic compounds. AB - When 56 flavonoids, 32 coumarins, five naphthoquinones, 12 anthraquinones and five structurally-related compounds were tested for their antimutagenic potencies with respect to mutagenicities induced by 2-nitrofluorene (2-NF), 3 nitrofluoranthene (3-NFA) and 1-nitropyrene (1-NP) in Salmonella typhimurium TA98 distinct structure-activity relationships were detected. First, the tetracyclic nitroarenes 3-NFA and 1-NP were in general more effectively antagonized by potent antimutagenic flavonoids and coumarins than the tricyclic 2-NF, while there were only minor differences with quinones. Secondly, antimutagenicity of natural compounds of plant origin correlated with the aglyconic nature 10 of a total of 15 glycosides were inactive, four flavonoid glycosides exerted antimutagenicity but to a distinctly lower degree than the corresponding aglycones. Thirdly, within flavonoids, coumarins and anthraquinones positive correlations were found between antimutagenic potency and the polarity of a molecule with the existence of an optimum of activity within flavonols and anthraquinones. However, polarity seemed to be unimportant within the chalcone and naphthoquinone series. Among flavonoids, the parent compounds flavone and flavanone were inactive, but all flavones and many flavonoids with phenolic hydroxyl groups exerted antimutagenicity. Antimutagenic potency reached a maximum with the presence of four hydroxyl functions-luteolin, kaempferol-though the position of hydroxyls was also a determinant of antimutagenic potency. Methylation of phenolic hydroxyl groups, however, always reduced antimutagenicity. A carbonyl group at carbon 4 was essential for antimutagenicity: two catechins and anthocyanidins each were inactive. On the other hand, ring C of the flavane nucleus was not essential for antimutagenicity: chalcones and dihydrochalcones were potent antimutagens. Among coumarins, the parent compound showed antimutagenicity against 1-NP and 3-NFA, although dihydrocoumarin, methylcoumarins and compounds with bulky substituents were inactive. Otherwise, antimutagenic activity depended on the presence of polar hydroxyl, amino or carboxyl groups at carbons 3, 4 or 7 but was diminished by interactions of hydroxyl groups vicinal to carbon 7. Again, antimutagenic potencies were reduced by alkylation or acetylation. Among furanocoumarins xanthotoxin exerted strong and bergapten moderate antimutagenicity, while psoralen (except against 3-NFA), isopimpinellin and the furanochromanones visnagin and khellin were inactive. Among anthraquinones, the principles delineated here were valid again, resulting in potent antimutagenicity of most phenolic compounds and inactivity of anthraquinone itself. Among compounds structurally related to anthraquinones, anthrone, acridone and xanthone exerted antimutagenicity, anthrone being the most potent one, while thioxanthone and 9 fluorenone were inactive. All naphthoquinones were potent antimutagens irrespective of the presence of methyl or hydroxyl functions. Plumbagin, 2-methyl 5-hydroxynaphthoquinone, however, showed exceptional antimutagenicity. PMID- 9207899 TI - Inhibition of the mutagenicity of 2-nitrofluorene, 3-nitrofluoranthene and 1 nitropyrene by vitamins, porphyrins and related compounds, and vegetable and fruit juices and solvent extracts. AB - When 21 vitamins including related compounds haemin, chlorophyllin, chlorophyll, biliverdin and bilirubin, as well as juices from five fruits and 25 vegetables and solvent extracts from the residues of fruits and vegetables were tested for their antimutagenic potencies with respect to mutagenicity induced by 2 nitrofluorene (2-NF), 3-nitrofluoranthene (3-NFA) and 1-nitropyrene(1-NP) in Salmonella typhimurium TA98 the following results were obtained. The tetracyclic nitroarenes 3-NFA and 1-NP were in general more effectively antagonized by potent antimutagenic compounds than the tricyclic 2-NF. beta-Carotene, retinol, retinal, retinoic acid, retinol palmitate, riboflavin 5'-phosphate, alpha-tocopherol, vitamins B12, C, K1 and K3 as well as biliverdin, bilirubin, chlorophyll, chlorophyllin and haemin exerted antimutagenicity against the nitroarenes cited previously. All other vitamins were inactive. While part of the juices were inactive, juices from cauliflower, carrots, chives, radishes and spinach exerted weak antimutagenic activities. However, weak to moderate co-mutagenic effects were seen with grapes, kiwi, pineapple, eggplant, celeriac, chicory greens, fennel leaves and radishes and strong effects with peppers which were not caused by the presence of growth-promoting factors. Most solvent fractions were inactive but fractions containing chlorophyll exerted antimutagenicity. PMID- 9207900 TI - Cytotoxicity and genotoxicity of diallyl sulfide and diallyl disulfide towards Chinese hamster ovary cells. AB - Two compounds found in garlic, diallyl sulfide (DAS) and diallyl disulfide (DDS), were tested for cytotoxic and genotoxic effects in a Chinese hamster ovary cell line. DDS was found to be more cytotoxic than DAS (showing a Dq of 1.6 micrograms/ml and a D0 of 0.6 microgram/ml as opposed to values of 295 and 90 micrograms/ml, respectively). Both compounds were found to induce both chromosome aberrations and sister chromatid exchanges (SCEs) with DDS again being more active on a weight-for-weight basis, exhibiting activity at concentrations below 10 micrograms/ml compared with the levels of 300 micrograms/ml and above required for DAS to show any effect. The addition of rat liver S-9 activation fraction to the assays modified the effects of the two compounds in a non-consistent manner. It reduced the induction of SCEs by both compounds, enhanced the generation of aberrations by DDS (but not by DAS) and radically altered the parameters of both survival curves, reducing the Dq values almost to zero but increasing the D0 values. PMID- 9207901 TI - Promotional effects of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) on N nitrosobis(2-oxopropyl)amine (BOP)-initiated carcinogenesis in hamsters. AB - The effects of administration of low doses of 4-(methylnitrosamino)-1-(3-pyridyl) 1-butanol (NNAL), a tobacco-specific nitrosamine, were investigated in hamsters treated with N-nitrosobis(2-oxopropyl)amine (BOP). Female Syrian golden hamsters were given a single sc injection of BOP at a dose of 10 mg/kg and then administered 2 or 5 ppm NNAL in their drinking water for 52 wk. Additional groups of animals received the BOP injection alone, or only the 2 or 5 ppm NNAL treatments as BOP-negative controls. At wk 53 of the experiment, all surviving animals were killed and the development of proliferative lesions was assessed histopathologically. The total incidence of combined carcinomatous and dysplastic lesions of the exocrine pancreas was significantly higher (P < 0.05) in the BOP/NNAL 5 ppm group than in the BOP alone group, although there was no statistically significant influence of NNAL on the development of either pancreatic adenocarcinomas or dysplastic lesions viewed singly. The treatments with NNAL alone did not induce any proliferative lesions of the exocrine pancreas. No significant intergroup differences were found in either incidence or multiplicity of islet cell proliferative lesions. Immunohistochemical examination of islet cell proliferative lesions (hyperplasias and adenomas) found in the BOP treated animals showed no significant differences in pancreatic hormone production between NNAL-treated and -untreated groups. The NNAL treatment did not exert any influence on lung, liver or kidney tumorigenesis. Thus, the results suggest that NNAL enhances BOP-induced exocrine but not endocrine pancreatic tumorigenesis in hamsters when given in the post-initiation phase. PMID- 9207902 TI - Chronic inhalation toxicity and oncogenicity of methyl methacrylate in rats and hamsters. AB - Male and female Fischer 344 rats were exposed to methyl methacrylate (MMA) monomer vapours at 0, 25, 100 and 400 ppm, 6 hr/day, 5 days/wk for 24 months and male and female Golden hamsters were exposed to similar vapour concentrations of MMA for 18 months. Parameters monitored throughout the study included clinical signs, individual body weights, haematology, clinical chemistry (rats only) and urinalyses (rats only). 10 rats per sex per exposure group were killed after 13 and 52 wk of exposure and all surviving rats were killed during wk 104-106. All surviving hamsters were killed at wk 78. Mortality and haematological, clinical chemistry and urinalyses parameters were not affected by MMA exposure. Body weights of male rats were not affected by exposure to MMA while body weights of female rats exposed to 400 ppm were lower than control values after wk 52. Male and female hamsters exposed to 400 ppm had body weight decreases ranging from 9 to 12% after wk 48. The nasal cavity was identified as the target organ for chronic toxicity in male and female rats exposed to 100 or 400 ppm. The microscopic nasal cavity changes occurred primarily in olfactory epithelium lining the dorsal meatus and consisted of degeneration of neuroepithelium, basal cell hyperplasia and atrophy of Bowman's glands. Hamsters did not have demonstrable nasal cavity microscopic changes. Chronic exposure to MMA vapour did not cause tumours in either rats or hamsters. PMID- 9207903 TI - Cutaneous xenobiotic metabolism: glycine conjugation in human and rat keratinocytes. AB - Glycine conjugation is an important route of metabolism and detoxication of carboxylic acids in the liver. In this paper the in vitro cutaneous metabolism of [carboxyl-14C]benzoic acid to its glycine conjugate hippuric acid in rat and human skin is reported. Cutaneous glycine conjugation was studied in F344 rat and human epidermal keratinocytes using two systems: (1) freshly isolated keratinocytes in suspension and (2) primary keratinocyte cultures. For comparative purposes, studies were also carried out in freshly isolated and cultured F344 rat hepatocytes. After incubation of 5 x 10(6) cells with 1 microM benzoic acid at 37 degrees C for 8 hr, no glycine conjugation was observed in rat and human keratinocyte suspensions, with greater than 98% of the radioactivity recovered as the parent compound. In contrast, cultured keratinocytes exhibited glycine conjugation, with 10.9 +/- 1.0% (mean SEM, n = 3) and 2.1 +/- 0.6% (mean SEM, n = 3) conversion to hippuric acid at 8 hr in rat and human cells, respectively. Tissue-specific differences in metabolism were observed, with conjugation in hepatocytes significantly greater (P < 0.05) than in keratinocytes at all times up to 8 hr. After incubation of benzoic acid with cultured hepatocytes for 8 hr, more than 98% of the of the radioactivity was recovered as the glycine conjugate. These studies indicate that rat and human skin possesses low, but demonstrable, glycine-conjugating activity, and that keratinocytes in primary culture may provide a better system than freshly isolated cell suspensions for studying such activity. PMID- 9207905 TI - Propeptin, a new inhibitor of prolyl endopeptidase produced by Microbispora. I. Fermentation, isolation and biological properties. AB - Propeptin, an inhibitor of the prolyl endopeptidase isolated from the mycelium of Microbispora sp. SNA-115, is an atypical cyclic peptide antibiotic. It was purified by column chromatographies on silica gel and Sephadex LH-20 and high performance liquid chromatography using an ODS column. Propeptin has the molecular formula of C113H142N26O27 and consists of nineteen amino acids. Propeptin inhibited prolyl endopeptidase of the genus Flavobacterium competitively when Z-Gly-Pro-pNA was used as a substrate. The inhibitor constant (Ki) was 0.70 microM. PMID- 9207904 TI - Skin sensitization thresholds: determination in predictive models. AB - For many years, test methods for the prospective identification of skin sensitizing chemicals have been widely available. However, although these techniques have permitted the identification of the great majority of skin sensitizers, their use in assessing the relative potency of a particular chemical as a human contact allergen has not been well described. A primary reason for this is the inherent difficulty of such an exercise. A complex phenomenon involving interactions between the vehicle, the allergen, the skin and its inflammatory responses takes place during the induction and elicitation of sensitization. All these factors can have a profound effect on the threshold values determined for a skin sensitizer. Consequently, whether the assessment is conducted in humans or in animal models, a threshold concentration is always a function of the method of measurement as much as the potency of the allergen. Although an exhaustive review has not been carried out, this paper considers the attempts that have been made to assess relative potency by the measurement of dose-response relationships and the determination of induction and elicitation thresholds in both animal models and in humans. The latter has special relevance for regulatory toxicology and this matter is given particular attention in this article. Finally, recommendations are made: (a) that threshold concentrations for skin sensitizers should be determined on a case by case basis in relation to the likely mode of skin contact; (b) where the data are used in comparisons of skin sensitization potency, then there should be standardization of the method used for the determinations. PMID- 9207906 TI - Lipohexin, a new inhibitor of prolyl endopeptidase from Moeszia lindtneri (HKI 0054) and Paecilomyces sp. (HKI-0055; HKI-0096). I. Screening, isolation and structure elucidation. AB - Lipohexin was isolated as a novel lipohexapeptide (I) (C39H68N6O9) from three fungal strains, Moeszia lindtneri HKI-0054, Paecilomyces sp. HKI-0055 and Paecilomyces sp. HKI-0096. The structure was elucidated by detailed mass spectrometric and NMR experiments. The proline-containing peptide displays moderate antibacterial activity against Bacillus subtilis ATCC 6633 and inhibits competitively the prolyl endopeptidase from human placenta. PMID- 9207907 TI - Lipohexin, a new inhibitor of prolyl endopeptidase from Moeszia lindtneri (HKI 0054) and Paecilomyces sp. (HKI-0055; HKI-0096). II. Inhibitory activities and specificity. AB - The new proline-containing lipohexapeptide lipohexin (I) isolated from three fungal strains, Moeszia lindtneri (HKI-0054) and Paecilomyces sp. (HKI-0055 and HKI-0096) is a competitive inhibitor of prolyl endopeptidase (PEP) from human placenta with IC50 of 3.5 microM. Specificity of lipohexin (I) is indicated by the much weaker inhibitory activity against bacterial prolyl endopeptidase from Flavobacterium meningosepticum (IC50 25 microM). No effect of lipohexin (I) was found on the activity of mechanistically related proteases such as proline specific proteases and other serine proteases. PMID- 9207908 TI - Biological activities of novel zaragozic acids, the potent inhibitors of squalene synthase, produced by the fungus, Mollisia sp. SANK 10294. AB - Four novel zaragozic acids, F-10863A, B, C and D, were isolated from a culture broth of the fungus Mollisia sp. SANK 10294. F-10863 compounds contain a 4,6,7 trihydroxy-2,8-dioxyobicyclo-[3.2.1]octane-3,4,5-tricarboxyl ic acid core like previously reported zaragozic acids, but the structures of the side chains are different. Recently, it was found that F-10863A is identical to zaragozic acid D3, while the other three are novel compounds. F-10863 compounds are potent inhibitors of squalene synthase like previously reported zaragozic acids, and, furthermore, they exhibit serum cholesterol-lowering activity in vivo. PMID- 9207909 TI - Semicochliodinol A and B: inhibitors of HIV-1 protease and EGF-R protein tyrosine kinase related to asterriquinones produced by the fungus Chrysosporium merdarium. AB - The known bisalkylated 2,5-dihydroxybenzoquinones didemethylasterriquinone D and isocochliodinol as well as the new metabolites semicochliodinol A and B have been isolated as inhibitors of HIV-1 protease from the culture broth of the fungus Chrysosporium merdarium P-5656. The structures were elucidated by spectroscopic methods. The NMR spectra of two compounds were completely assigned. The metabolites inhibit HIV-1 protease with an IC50 value as low as 0.17 microM and epidermal growth factor receptor protein tyrosine kinase at 15 to 60 microM and are therefore valuable lead compounds for these targets. Molecular modelling of the HIV-1-protease-inhibitor complexes showed hydrogen bonding between the dihydroxybenzoquinone moiety of didemethylasterriquinone D and isocochliodinol to both active-site aspartic acids (Asp25/Asp25') of the protease and the indole parts of the inhibitors filling the P2 and P2' pockets of the protease. PMID- 9207910 TI - Cyclothialidine analogs, novel DNA gyrase inhibitors. AB - DNA gyrase inhibitors, cyclothialidines B, C, D and E were isolated from four Streptomycete strains (NR 0659, NR 0660, NR 0661 and NR 0662). Their structures have been elucidated based on the amino acid analysis of the hydrolysates, NMR and HRFAB-MS experiments and shown to be cyclothialidine analogs. The absolute stereochemistry has been determined by the chiral HPLC analysis of the hydrolysates. Cyclothialidines B, D and E are novel and potent inhibitors of DNA gyrase. PMID- 9207911 TI - The effect of carbon source, temperature and aeration on the production of ascosteroside, a novel antifungal agent, by Ascotricha amphitricha. AB - This paper describes the optimization of production of ascosteroside, a novel antifungal agent with an alpha-linked glycoside of a lanosterone-type triterpenoid structure. Glucose, sorbose and inositol were determined to be the best carbon sources for the production of ascosteroside. Temperature affected levels of ascosteroside, with production being highest at 16 degrees C with 1% glucose, and lowest at 32 degrees C. Dissolved oxygen levels were found to be critical in the production of ascosteroside in fermenter cultures. In order for production of ascosteroside to occur in fermenter cultures, the threshold level of dissolved oxygen was found to be above 26%. PMID- 9207913 TI - Kinetic isotope effect and reaction mechanism of 2-deoxy-scyllo-inosose synthase derived from butirosin-producing Bacillus circulans. AB - The mechanism of 2-deoxy-scyllo-inosose synthase reaction, a carbocycle formation step from D-glucose-6-phosphate in the biosynthesis of the 2-deoxystreptamine aglycon of clinically important aminocyclitol antibiotics, was investigated with a partially purified enzyme from butirosin-producing Bacillus circulans SANK 72073, Nonlabeled and double-labeled D-[4-2H, 3-15O]glucose-6-phosphate were used for cross-over experiment, and the oxime-TMS ether derivative of the 2-deoxy scyllo-inosose product was analyzed by GC-MS. The deuterium label at C-4 of the substrate appeared to be retained at C-6 of the inosose product without scrambling of the double-labeled isotopes. Since the transient reduction of NAD+ cofactor was proved to be essential in the 2-deoxy-scyllo-inosose reaction, the hydride abstraction and returning appeared to take place within the same glucose molecule. The observed kinetic isotope effect was estimated to be kH/kD = 2.4. These results strongly suggest that this carbocycle formation is catalyzed by a single 2-deoxy-scyllo-inosose synthase enzyme with catalytic requirement of NAD+, the mechanism of which appears to be resembled closely to the 2-deoxy-scyllo inosose synthase in the Streptomyces fradiae. PMID- 9207912 TI - Chemical and biological characterization of two FK506 analogs produced by targeted gene disruption in Streptomyces sp. MA6548. AB - Two genetically engineered mutant strains of Streptomyces sp. MA6548 produced two FK506 analogs, 9-deoxo-31-O-demethylFK506 and 31-O-demethylFK506. The structures were determined by a combination of NMR and mass spectrometry. These compounds exhibited immunosuppressive and antifungal activities, albeit reduced, compared to FK506. Both compounds contain a free hydroxyl group at C-31 for the synthesis of novel FK506 derivatives. PMID- 9207914 TI - Synthesis and structure-activity relationships of a novel oral carbapenem, CS 834. AB - We have studied an ester prodrug of a carbapenem to develop a potent orally active beta-lactam antibiotic. A variety of 1 beta-methylcarbapenem derivatives have been synthesized. We have found that some derivatives having an amide group in the C-2 side chain show potent and well balanced antibacterial activities as well as high stability against dehydropeptidase-I. Oral absorption of derivatives has been optimized by modifying the C-3 ester promoiety. Pivaloyloxymethyl (1R, 5S, 6S)-6-[(R)-1-hydroxyethyl]-l-methyl-2-[(R)-5-oxopyrrolidin-3-yl thio]- l carbapen-2-em-3-carboxylate, CS-834, has been selected as the most promising compound for further evaluation. PMID- 9207915 TI - Cytovaricin B, a new inhibitor of JAK-STAT signal transduction produced by Streptomyces torulosus. PMID- 9207916 TI - Fusarin C, (7Z)-fusarin C and (5Z)-fusarin C; inhibitors of dihydroxynaphthalene melanin biosynthesis from Nectria coccinea (Cylindrocarpon sp.). PMID- 9207917 TI - Stereospecific synthesis of a 1 beta-methylcarbapenem intermediate. PMID- 9207918 TI - Thiomarinols D, E, F and G, new hybrid antimicrobial antibiotics produced by a marine bacterium; isolation, structure, and antimicrobial activity. PMID- 9207919 TI - Syntheses and absolute structures of novel protein farnesyltransferase inhibitors, kurasoins A and B. PMID- 9207920 TI - Growth stimulation of colorectal carcinoma cells via the c-kit receptor is inhibited by TGF-beta 1. AB - Activation of the receptor tyrosine kinase c-kit by the kit-ligand, also known as stem cell factor (SCF), is essential to melanocyte and germ cell development and during the early stages of hematopoiesis. Deregulated expression of c-kit has been reported in malignancies affecting these lineages, i.e., myeloid leukemias, melanomas, and germ cell tumors. In addition, c-kit and SCF are coexpressed in some breast and colorectal cancer (CRC) cells, raising the question of whether c kit serves an autocrine role in normal or malignant epithelial tissues. In this study, we demonstrate that human colorectal carcinomas, but not normal colorectal mucosa cells, coexpress SCF and c-kit in situ. Expression of c-kit was also observed in mucosa adjacent to colorectal tumor tissue. Consistent with a growth regulatory role of SCF in CRC cells, exogenous SCF stimulated anchorage-dependent and anchorage-independent growth in four out of five CRC cell lines. Exogenous transforming growth factor (TGF)-beta 1 added at nanomolar concentrations to HT 29 CRC cells, which express the type I, II, and III TGF-beta receptors, downregulated c-kit expression to background levels and inhibited c-kit-dependent proliferation. Similarly, TGF-beta 1 inhibited SCF-dependent proliferation of three first-passage CRC cell lines. In summary, expression of the potential autocrine SCF/ c-kit axis is a tumor-associated phenomenon in colorectal cancer that can be suppressed by TGF-beta 1 in TGF-beta-responsive CRC cells. PMID- 9207921 TI - Nitric oxide inhibits neutrophil beta 2 integrin function by inhibiting membrane associated cyclic GMP synthesis. AB - The aim of this investigation was to identify the mechanism by which nitric oxide inhibits neutrophil beta 2 integrin dependent adherence. Isolated rat neutrophils from blood and peritoneal exudates were exposed for 2 min to nitric oxide generated by diethylamine-NO at rates between 1.6 and 138 nmol/min. Exposure to nitric oxide at rates less than 14 nmol/min had no effect on adherence. Exposure to 14 to 56 nmol nitric oxide/min inhibited beta 2 integrin dependent adherence to endothelial cells, nylon columns, and fibrinogen-coated plates, but higher concentrations had no significant effect on adherence. Adherence by beta 2 integrins could be restored by incubating cells with dithioerythritol, phorbol 12 myristate 13-acetate, or 8-bromo cyclic GMP. Elevations in cellular cyclic GMP concentration were associated with adherence, but this did not occur after cells were exposed to concentrations of nitric oxide that inhibited beta 2 integrin dependent adherence. Elevations in cyclic GMP did occur after cells were incubated with dithioerythritol or phorbol 12-myristate 13-acetate. Concentrations of nitric oxide that inhibited beta 2 integrin-dependent adherence also inhibited catalytic activity of membrane associated guanylate cyclase and binding of atrial natriuretic peptide, but were insufficient to activate cytosolic guanylate cyclase. Nitric oxide did not inhibit neutrophil oxidative burst or degranulation, nor effect beta 2 integrin expression or adherence that did not depend on beta 2 integrins, nor cause oxidative stress identified in terms of cellular glutathione concentration or protein nitrotyrosine. The results indicate that nitric oxide inhibited beta 2 integrins in a concentration dependent fashion by inhibiting cell-surface transduction of signals linked to the activity of membrane-bound guanylate cyclase. The inhibitory effect could be overcome by providing cells with cyclic GMP exogenously or by stimulating cytosolic guanylate cyclase. PMID- 9207922 TI - Apoptotic cell death in neuronal differentiation of P19 EC cells: cell death follows reentry into S phase. AB - Apoptotic cell death was observed during aggregate culture of the mouse embryonal carcinoma cell line P19 exposed to all-trans retinoic acid (tRA). This finding was confirmed by genomic DNA agarose gel electrophoresis and transmission electron microscopy. Apoptosis was associated with P19 cell neuronal differentiation; alternative causes of cell death, i.e., cavitation-related, cytotoxicity of tRA, or spontaneous cell death were excluded. Analysis by flow cytometry revealed that the apoptosis was likely to occur in multiplying cells that underwent to reentering into S phase. We therefore examined 5-bromo-2' deoxyuridine (BrdU) incorporation and proliferating cell nuclear antigen (PCNA) expression and localization in the aggregates by immunofluorescent staining. Although the P19 cells in the aggregates exposed to tRA incorporated BrdU at an equivalent level to those not exposed to tRA, the cells showed diminished PCNA expression and nuclear accumulation. We propose that P19 apoptosis during neuronal differentiation is a model system in which programmed cell death occurs simultaneously with cell division leading to differentiation. PMID- 9207923 TI - Human umbilical vein endothelial cells express high affinity receptors for factor Xa. AB - The binding of [125I]-factor Xa to human umbilical vein endothelial cell (HUVEC) monolayers was studied. At 7 degrees C, [125I]-factor Xa bound to a single class of binding sites with a dissociation constant value of 6.6 +/- 0.8 nM and a binding site density of 57,460 +/- 5,200 sites/cell (n = 3). Association and dissociation kinetics were of a pseudo-first order and gave association and dissociation rate constant values of 0.15 x 10(6) M-1 s-1 and 4.0 x 10(-4) s-1, respectively. [125I]-factor Xa binding was inhibited by factor Xa but was not affected by factor X, thrombin or monoclonal antibodies against factor V, antithrombin-III or tissue factor pathway inhibitor (TFPI) but was inhibited by an antibody specific for the effector cell protease receptor-1 (EPR-1), a well known receptor of factor Xa on various cell types. [125I]-factor Xa binding to HUVEC was not affected by various inhibitors of factor Xa such as DX 9065, pentasaccharide-antithrombin-III or TFPI. Factor Xa increased intracellular free calcium levels and phosphoinositide turnover in endothelial cells and, when added to HUVEC in culture, factor Xa was a potent mitogen, stimulating an increase in cell number at a 0.3 to 100 nM concentration. HUVEC-bound factor Xa promoted prothrombin activation in the presence of factor Va only. This effect was inhibited by both indirect and direct inhibitors of factor Xa. These findings indicate that HUVEC express functional high affinity receptors for factor Xa, related to EPR-1, which may be of importance in the regulation of coagulation and homeostasis of the vascular wall. PMID- 9207924 TI - Distribution of HSP70, protein kinase C, and spectrin is altered in lymphocytes during a fever-like hyperthermia exposure. AB - Many B and T lymphocytes display a significant heterogeneity with respect to the subcellular distribution of the cytoskeletal protein spectrin and protein kinase C (PKC), both of which often can be found in a large cytoplasmic aggregate in these cell types. In addition to spectrin and PKC, we recently have reported that HSP70 is also a component of this lymphocyte aggregate. Moreover, these three proteins can undergo dynamic and reversible changes in their localization causing "assembly" of the aggregate in response to various conditions associated with lymphocyte activation, indicating that this naturally occurring aggregate structure is sensitive to activation status. We show here that the same changes in HSP70/spectrin/PKC localization induced by PKC activation also can be caused, in vitro and in vivo, by a mild hyperthermia exposure, as occurs during a natural fever (39.5-40 degrees C, 2-12 hr). This mild heat exposure also triggers the activation of PKC, a major heat shock response, and lymphocyte proliferation. The increase in PKC activity, HSP70-spectrin-PKC aggregate formation, and heat shock protein expression resulting from exposure to fever-like hyperthermia are all inhibited by calphostin C, a specific inhibitor of PKC. These data demonstrate that changes observed during lymphocyte activation could be induced by a mild hyperthermia exposure occurring during a normal febrile episode. PMID- 9207925 TI - Insulin-like growth factor-I mediates osteoclast-like cell formation stimulated by parathyroid hormone. AB - There have been several lines of evidence that parathyroid hormone (PTH) stimulates production of insulinlike growth factor I (IGF-I) in bone and that IGF I stimulates osteoclast formation. Thus, the present study was performed to clarify the possible role of IGF-I in PTH-stimulated osteoclastlike cell formation and the role of PTH-responsive dual signal transduction systems (cyclic [c] AMP-dependent protein kinase [PKA] and calcium/protein kinase C [PKC]) in its mechanism. Treatment with anti-IGF-I antibody (1-10 micrograms/ml) partially but significantly blocked hPTH-(1-34)-stimulated osteoclastlike cell formation in unfractionated mouse bone cell cultures, although it did not affect osteoclastlike cell formation stimulated by 1,25-dihydroxyvitamin D3. Rp-cAMP5 (10(-4) M), a direct PKA inhibitor, as well as two types of PKC inhibitors, H-7 (10 microM) and staurosporine (3 nM), and dantrolene (10(-5) M), an inhibitor of calcium mobilization from intracellular calcium stores, all significantly blocked PTH-stimulated osteoclastlike cell formation. Anti-IGF-I antibody (3 micrograms/ml) significantly blocked osteoclastlike cell formation stimulated by 10(-4) M dbcAMP, 10(-4) M Sp-cAMPS, a direct PKA activator, and 10(-5) M forskolin in mouse bone cell cultures. Dibutyryl cAMP, forskolin, and hPTH-(1-34) significantly stimulated mRNA expression of both IGF-I and IGF-binding protein 5 (IGFBP-5) in these cultures, but neither 10(-7) M PMA, a PKC activator, nor 10( 7) M A23187 did. Moreover, anti-IGF-I antibody significantly blocked osteoclastlike cell formation stimulated by the conditioned medium from MC3T3-E1 cells pretreated with 10(-8) PTH-(1-34), which induced IGF-I and IGFBP-5 mRNA expression in these cells. In conclusion, the present study indicates that IGF-I mediates osteoclastlike cell formation stimulated by PTH and that the PKA pathway is involved in its mechanism. However, IGF-I does not seem to be the sole effector molecule to be active in this system. PMID- 9207926 TI - Cysteine proteinases are responsible for characteristic transketolase alterations in Alzheimer fibroblasts. AB - Cultured fibroblasts from patients affected by Alzheimer's disease (AD) exhibited peculiar alterations of the enzyme transketolase (TK). Abnormalities (dubbed alkaline bands, ab) consisted of enzyme forms having unusually high pl and were proposed as a marker of the disease in living patients. The mechanisms of TK-ab expression were investigated with the use of cysteine proteinase inhibitors and purified preparations of either rat liver or human cysteine proteinases. The cysteine proteinase inhibitors N-acetyl-leu-leu-norleucinal (ALLN), L-trans-Epoxy succinyl-leucylamido(4-guanidino)butane (E-64), and egg white cystatin added to AD cells just prior to extraction abolished TK abnormalities. Moreover, 1 day incubation of AD cultures with either ALLN (10 micrograms/ml), NH4Cl (10 mM), or KCl (30 mM) prevented TK-ab generation, due, presumably, to an impairment of lysosomal functions. Isolated rat liver cysteine proteinases were able to degrade TK in normal extracts and reproduce the characteristic TK-ab of AD fibroblasts. Moreover, pure human cathepsin H was also shown to partially induce an Alzheimer like TK pattern and cleave normal TK to a 35 kDa fragment as spontaneously occurring in AD fibroblasts. The explanation of mechanisms of TK-ab formation provided evidence for an underlying imbalance of proteolysis in AD fibroblasts due to a relative increase/derangement of the cysteine proteinases cathepsins which might be also involved in the reported abnormal processing of multiple cellular components. PMID- 9207928 TI - Osteoclast activation: potent inhibition by the bisphosphonate alendronate through a nonresorptive mechanism. AB - Alendronate, an aminobisphosphonate used in the treatment of osteoporosis, is a potent inhibitor of bone resorption. Its mechanism of action is unknown. Because it localizes to bone surfaces, we compared the sensitivity of components of the resorptive process to incubation on alendronate-coated bone surfaces. We found that bone resorption by osteoclasts isolated from neonatal rat bone was unaffected by alendronate (10(-4) M). Osteoclast production in bone marrow cultures, as assessed by the production of calcitonin-receptor positive cells, was observed even at 10(-4) M, but bone resorption in these cultures was almost completely abolished by 10(-5) M alendronate. The greater sensitivity of osteoclast activation to inhibition by alendronate that these results suggest was supported by similar inhibition of osteoblast-mediated activation of osteoclasts from neonatal rat bone. Thus, activation of osteoclasts by osteoblastic/stromal cells is apparently the most sensitive component of the pathway whereby bone resorption is affected. Moreover, the ability of alendronate to suppress osteoclastic activation does not depend on resorption-mediated release of alendronate from bone surfaces. This ability extends the range of cell types and processes that might be affected by alendronate, beyond those in the immediate vicinity of resorbing cells, to include any cell that comes into contact with alendronate-coated bone surfaces. PMID- 9207927 TI - Mechanisms of inhibition by heparin of PDGF stimulated MAP kinase activation in vascular smooth muscle cells. AB - Heparin and heparan are potent inhibitors of vascular smooth muscle cell (VSMC) proliferation. To investigate the mechanisms by which heparin suppresses growth factor stimulated mitogenesis, the present experiments investigated the effects of heparin on platelet-derived growth factor (PDGF) stimulated signal transduction pathways. Heparin treatment substantially inhibited PDGF-BB stimulated rat VSMC growth. Western analysis showed a 30 min PDGF-BB treatment of VSMC induced the tyrosine phosphorylation of multiple protein bands; cotreatment with heparin inhibited mitogen-activated protein (MAP) kinase tyrosine phosphorylation but had little effect on PDGF receptor tyrosine phosphorylation. In-gel kinase assays demonstrated that heparin inhibited PDGF-BB stimulated MAP kinase activity at late (25 min) but not early (10 min) time points. These data indicate that heparin does not inhibit the initial signalling events after PDGF BB binding but instead acts through an alternate mechanism to inhibit MAP kinase. To investigate if heparin directly stimulates tyrosine phosphatase-mediated suppression of MAP kinase, we treated VSMC with orthovanadate, a tyrosine phosphatase inhibitor. Heparin inhibited MAP kinase tyrosine phosphorylation after orthovanadate treatment, indicating that heparin does not suppress MAP kinase by enlistment of a tyrosine phosphatase. Experiments were performed to investigate signalling pathways upstream of MAP kinase. To determine if protein kinase C (PKC) mediates PDGF-BB, serum, and EGF stimulation of MAP kinase, we treated VSMC overnight with phorbol ester (PMA) to downregulate PKC. Abolition of conventional and novel PKC activity significantly suppressed both serum and PDGF BB induced MAP kinase activation, indicating protein kinase C is an important mediator for these mitogens. In contrast, downregulation of these PKC isoforms had little effect on EGF stimulation of MAP kinase. As heparin inhibits PDGF and serum but not EGF stimulation of MAP kinase, there data precisely correlate heparin inhibition of MAP kinase with activation through PKC-dependent pathways. Immunoprecipitation analysis found that heparin inhibited serum, PMA, and PDGF but not EGF induced raf-1 phosphorylation. These studies demonstrate that heparin did not block PDGF-BB receptor activation, which initiates the mitogenic signalling cascade. Heparin did inhibit specific postreceptor second messenger signals, such as the late phase activation of MAP kinase, which may be mediated by suppression of PKC-dependent pathways. PMID- 9207929 TI - Decreased serglycin proteoglycan size is associated with the platelet alpha granule storage defect in Wistar Furth hereditary macrothrombocytopenic rats. Serglycin binding affinity to type I collagen is unaltered. AB - The Wistar Furth (WF) rat has a hereditary defect in platelet formation that resembles gray platelet syndrome of man with a large mean platelet volume and platelet alpha granule deficiency. The alpha granule abnormality is suggestive of a defect in granule packaging and/or stability. Proteoglycans are hypothesized to play a role in granule packaging. Therefore, we have analyzed the structure of the platelet proteoglycan, serglycin, in platelets of WF and normal Wistar rats. Normal and Wistar Furth rats were injected with 35S-sulfate to label platelet proteoglycans via synthesis by the megakaryocytes, and platelets were isolated 3 days later. We found that WF rat platelets have only one-third of the normal proteoglycan mass per unit platelet volume, and the proteoglycans are smaller in hydrodynamic size with shorter glycosaminoglycan chains than those of Wistar rats. However, WF rat platelet proteoglycans showed no defect in binding to collagen on affinity coelectrophoresis gels. We conclude that the structure of WF rat platelet proteoglycans is abnormal, and speculate that this abnormality may contribute to abnormal packaging of the alpha granule contents. Leakage of alpha granule contents into the marrow by platelets and megakaryocytes could perturb the marrow matrix, and promote the development of myelofibrosis noted in gray platelet syndrome. PMID- 9207930 TI - Inhibition of neutrophil oxidative burst and granule secretion by wortmannin: potential role of MAP kinase and renaturable kinases. AB - Exposure of neutrophils to a variety of agonists including soluble chemoattractant peptides and cytokines results in degranulation and activation of the oxidative burst (effector functions) that are required for bacterial killing. At present, the signaling pathways regulating these important functions are incompletely characterized. Mitogen-activated protein (MAP) kinases (MAPK) as well as members of a family of "renaturable kinases" are rapidly activated in neutrophils in response to diverse physiological agonists, suggesting that they may regulate cell activation. Antagonists of phosphatidyl inositol-3-(OH) kinase (PI3-kinase) such as wortmannin (Wtmn) inhibit these effector responses as well as certain of the above-mentioned kinases, leading to the suggestion that these enzymes lie downstream of PI3-kinase in the pathway regulating the oxidative burst and granule secretion. However, an apparent discrepancy exists in that, while virtually obliterating activity of PI3-kinase and the oxidase at low concentrations (ID50 < 20 nM), Wtmn has only variable inhibitory effects on MAPK even at substantially higher concentrations (75-100 nM). This raises the possibility that the inhibitory effects of Wtmn are mediated via other enzyme systems. The purpose of the current study was therefore to compare the effects of Wtmn on PI3-kinase activity and on the chemoattractant-activated kinases, and to determine the potential relationship of these pathways to microbicidal responses. In human neutrophils, both the oxidative burst and granule secretion induced by fMLP were inhibited by Wtmn but at markedly different concentrations: the oxidative burst was inhibited with an ID50 of < 5 nM while granule secretion was only partially inhibited at concentrations exceeding 75 nM. Activation of both MEK-1 and MAPK in response to fMLP was only partially inhibited by high doses of Wtmn (ID50 of > 100 nM and approximately 75 nM, respectively). In contrast, Wtmn potently inhibited fMLP-induced activation of the 63 and 69 kDa renaturable kinases (ID50 approximately 5-10 nM). We speculate that the renaturable kinases may be involved in the regulation of the oxidative burst, whereas the MAPK pathway may play a role in other neutrophil functions such as granule secretion. PMID- 9207931 TI - Disruption of integrin alpha 5 beta 1 signaling does not impair PDGF-BB-mediated stimulation of the extracellular signal-regulated kinase pathway in smooth muscle cells. AB - Smooth muscle cell (SMC) proliferation is dependent on both anchorage to the extracellular matrix by integrins and the presence of growth factors. Integrins and growth factor receptors transduce signals that seem to converge on the extracellular signal-regulated (ERK) pathway, but the molecular basis for this interaction is not known. SMC proliferation has previously been shown to be supported by culture on fibronectin (FN), whereas cells cultured on laminin (LN) are growth inhibited. In the present study, we examined the mitogenic response to platelet-derived growth factor BB (PDGF-BB) in baboon SMCs cultured on FN vs. LN. Induction of DNA synthesis and the activity of ERK and the ERK activating kinase MKK-1 were reduced only slightly after stimulation with PDGF-BB in cells cultured on LN vs. those cultured on FN. We tested the possibility that endogenous FN secretion contributes to the ability of the cells to respond to PDGF stimulation during culture on LN. Inhibition of interactions between FN and integrin alpha 5 beta 1 by the competitive GRGDSP-peptide or anti-alpha 5 integrin antibody restricted cell spreading, reduced cell-surface staining for alpha 5 beta 1 and FN fibrils, and inhibited PDGF-BB-induced DNA synthesis. These results showed that SMC growth on LN required a provisional FN matrix. Although disruption of interactions between alpha 5 beta 1 and FN by the GRGDSP-peptide prevented PDGF BB-induced DNA synthesis, neither ERK activity nor translocation of ERKs into the nucleus was inhibited. These results show that integrins regulate SMC growth through pathways that function in parallel with, but distinct from, growth factor mediated ERK signaling. PMID- 9207932 TI - Mutation associated with Crouzon syndrome causes ligand-independent dimerization and activation of FGF receptor-2. AB - FGF signaling is clearly important for proper bone development, and several autosomally dominant forms of genetic bone disorders have been mapped to FGF receptors 1, 2, and 3. We have studied the biological effects of the most commonly mutated cysteine residue in FGFR-2 which is detected in individuals with Crouzon syndrome, an autosomally dominant trait which causes premature fusion of the skull bones (craniosynostosis). This Crouzon mutation replaces the cysteine at position 342 with tyrosine, thus disrupting the formation of the third immunoglobulin (Ig)-like loop in the extracellular portion of the receptor. By transfecting mutated and wild-type receptors into a variety of cell lines, we have shown that the C342Y mutation in FGFR-2 produces a receptor which is constitutively activated and capable of transforming NIH3T3 cells and preventing the differentiation of C2 myoblasts in the absence of ligand. Constitutive activation appears to result from the ability of this receptor to form stable interreceptor dimers which involve disulfide bonds between the remaining free cysteine in the mutant receptor. The altered conformation of the third Ig-like domain in the mutated receptor also results in a drastically reduced ability to bind FGF-1 or FGF-2 and in a reduced level of receptor glycosylation. Thus it appears that Crouzon syndrome results from constitutive activation of FGFR-2 and that uncontrolled FGF signaling produces alterations of intramembranous bone development and premature closing of cranial sutures. PMID- 9207933 TI - Cytoskeletal association of epidermal growth factor receptor and associated signaling proteins is regulated by cell density in IEC-6 intestinal cells. AB - Epidermal growth factor (EGF) mediates a variety of physiologic responses in rat intestine. EGF receptor (EGFR) responsiveness to EGF is mediated by the surface expression of high affinity EGFR, which is associated with the cytoskeleton (CSK). EGFR signal transduction appears to be mediated by the CSK association of EGFR and related signaling proteins. In the nontransformed intestinal cell line IEC-6, expression of EGFR, Src homology and collagen protein (SHC), phospholipase C gamma 1 (PLC gamma), and their tyrosine phosphorylation in response to EGF was assayed by immunoblot. The distribution of EGFR and tyrosine-phosphorylated EGFR was regulated by cell density. At confluence, EGFR and tyrosine-phosphorylated EGFR were predominantly associated with the Triton X-100-insoluble CSK at confluence, while predominantly Triton X-100-soluble at subconfluence. PLC gamma was predominantly soluble at both states of confluence. Confluent but not subconfluent IEC-6 cells demonstrated a cascade of EGF-mediated events consisting of a transient CSK association of PLC gamma with EGFR, a brief expression of tyrosine-phosphorylated PLC gamma, a brief increase in PLC gamma CSK association, and a prolonged soluble association of PLC gamma with the EGFR. EGF led to an increase in the CSK association of SHC at both states of confluence and was greater at confluence. EGFR association with SHC was primarily soluble at subconfluence, while at confluence EGFR association was markedly increased and predominantly in the CSK. Thus, cell density regulates the CSK association of the EGFR and its ability to associate and activate signaling pathways in intestinal cells. PMID- 9207934 TI - Pyridazinodiazepines as a high-affinity, P2-P3 peptidomimetic class of interleukin-1 beta-converting enzyme inhibitor. PMID- 9207935 TI - Targeting nitric oxide (NO) delivery in vivo. Design of a liver-selective NO donor prodrug that blocks tumor necrosis factor-alpha-induced apoptosis and toxicity in the liver. AB - We have designed a drug that protects the liver from apoptotic cell death by organ-selective pharmacological generation of the bioregulatory agent, nitric oxide (NO). The discovery strategy involved three steps: identifying a diazeniumdiolate ion (R2N[N(O)NO]-, where R2N = pyrrolidinyl) that spontaneously decomposes to NO with a very short half-life (3 s) at physiological pH; converting this ion to a series of potential prodrug derivatives by covalent attachment of protecting groups that we postulated might be rapidly removed by enzymes prevalent in the liver; and screening the prodrug candidates in vitro and in vivo to select a lead and to confirm the desired activity. Of five cell types examined, only cultured hepatocytes metabolized O2-vinyl 1-(pyrrolidin-1 yl)diazen-1-ium-1,2-diolate (V-PYRRO/NO) to NO, triggering cyclic guanosine 3',5' monophosphate (cGMP) synthesis and protecting the hepatocytes from apoptotic cell death induced by treatment with tumor necrosis factor-alpha (TNF alpha) plus actinomycin D. In vivo, V-PYRRO/NO increased liver cGMP levels while minimally affecting systemic hemodynamics, protecting rats dosed with TNF alpha plus galactosamine from apoptosis and hepatotoxicity. The results illustrate the potential utility of diazeniumdiolates for targeting NO delivery in vivo and suggest a possible therapeutic strategy for hepatic disorders such as fulminant liver failure. PMID- 9207936 TI - Structure-activity relationships of N-hydroxyurea 5-lipoxygenase inhibitors. AB - The discovery of second generation N-hydroxyurea 5-lipoxygenase inhibitors was accomplished through the development of a broad structure-activity relationship (SAR) study. This study identified requirements for improving potency and also extending duration by limiting metabolism. Potency could be maintained by the incorporation of heterocyclic templates substituted with selected lipophilic substituents. Duration of inhibition after oral administration was optimized by identification of structural features in the proximity of the N-hydroxyurea which correlated to low in vitro glucuronidation rates. Furthermore, the rate of in vitro glucuronidation was shown to be stereoselective for certain analogs. (R)-N [3-[5-(4-Fluorophenoxy)-2-furyl]-1-methyl-2-propynyl]-N-hydroxyure a (17c) was identified and selected for clinical development. PMID- 9207938 TI - Pyrrolomorphinans as delta opioid receptor antagonists. The role of steric hindrance in conferring selectivity. AB - A series of 2',3'-disubstituted pyrrolomorphinans (5a-i) were synthesized to determine the role of steric hindrance at mu and kappa receptors in promoting delta opioid receptor antagonist selectivity. In smooth muscle preparations, five members of the series (5a-c,e,f) possessed Ke values in the range 2-15 nM and were delta selective. Since the unsubstituted analogue 4 possessed delta antagonist potency of similar magnitude, but was not delta selective, it is suggested that the 2',3'-substitution confers delta selectivity by hindering the interaction of the pharmacophore at mu and kappa receptors, while not affecting delta receptors. PMID- 9207937 TI - Zinc ejection as a new rationale for the use of cystamine and related disulfide containing antiviral agents in the treatment of AIDS. AB - The highly conserved and mutationally intolerant retroviral zinc finger motif of the HIV-1 nucleocapsid protein (NC) is an attractive target for drug therapy due to its participation in multiple stages of the viral replication cycle. A literature search identified cystamine, thiamine disulfide, and disulfiram as compounds that have been shown to inhibit HIV-1 replication by poorly defined mechanisms and that have electrophilic functional groups that might react with the metal-coordinating sulfur atoms of the retroviral zinc fingers and cause zinc ejection. 1H NMR studies reveal that these compounds readily eject zinc from synthetic peptides with sequences corresponding to the HIV-1 NC zinc fingers, as well as from the intact HIV-1 NC protein. In contrast, the reduced forms of disulfiram and cystamine, diethyl dithiocarbamate and cysteamine, respectively, were found to be ineffective at zinc ejection, although cysteamine formed a transient complex with the zinc fingers. Studies with HIV-1-infected human T cells and monocyte/macrophage cultures revealed that cystamine and cysteamine possess significant antiviral properties at nontoxic concentrations, which warrant their consideration as therapeutically useful anti-HIV agents. PMID- 9207939 TI - Probing the active sites of monoamine oxidase A and B with 1,4-disubstituted tetrahydropyridine substrates and inactivators. AB - As part of our efforts to characterize more fully the structural features of the monoamine oxidase (MAO) A and B active sites, we have examined the substrate and inhibitor properties of several 1-methyl- and 1-cyclopropyl-4-aryl-1,2,3,6 tetrahydropyridine derivatives with the human placental A and beef liver B forms of the enzyme. We find that the 4-(2-phenylphenyl) analog 23 exhibits a high activity and selectivity for MAO-A while the 4-(3-phenylphenyl) analog 22 shows activity only with MAO-B. Selectivities similar to those of the N-methyl series are observed with a series of N-cyclopropyl mechanism based inactivators. These results support a topological analysis which attempts to identify steric factors related to the reported substrate and inhibitor selectivities of these two flavoproteins and provide a better definition of the size of the active sites of the two enzymes. PMID- 9207940 TI - Conformationally restrained melatonin analogues: synthesis, binding affinity for the melatonin receptor, evaluation of the biological activity, and molecular modeling study. AB - The design, synthesis, and biological profile of several indole melatonin analogues with a conformationally restricted C3 amidoethane side chain are presented. Examination of the accessible conformations of the melatonin side chain led us to explore some of its fully or partially restricted analogues, 2 12, the binding affinity values of which were utilized to gain further insight on the melatonin binding site. Two pharmacophoric models have been devised for melatonin and the active compounds by conformational analysis and superimposition performed using the DISCO program. In these models, the melatonin side chain can adopt a gauche/anti conformation out of the indole plane. Another contribution of this study regards the observation of a possible binding point interaction around the C2 position of the indole, as suggested by the remarkably increased binding affinity observed in the C2-substituted analogues 6 and 9 and especially in the more rigid analogue 5. The biological activity and the efficacy of the new compounds were tested by measuring the inhibition of the forskolin-stimulated cAMP accumulation and the GTP gamma S index. Both analyses demonstrated that all of the compounds were full agonists with the exception of 4 and 9, which showed a slight reduction in efficacy and would seem to be partial agonists. PMID- 9207941 TI - 1-(2-Alkanamidoethyl)-6-methoxyindole derivatives: a new class of potent indole melatonin analogues. AB - A new series of indole melatonin analogues, bearing the amido ethyl side chain attached at the N-1 position of the indole nucleus, were synthesized and tested for their affinity for the melatonin receptor isolated from quail optic tecta in a series of in vitro ligand-binding experiments using 2-[125I]iodomelatonin as the labeled ligand. The biological activity was evaluated using two models: effects on the forskolin-stimulated cAMP accumulation in explants from quail optic tecta and evaluation of the GTP gamma S index derived from competition experiments performed in the absence or presence of GTP gamma S. Compounds 2a and 2k-n, obtained by shifting the methoxy group and the ethylamido side chain from the C-5 and C-3 positions of melatonin to the C-6 and N-1 positions of the indole nucleus, exhibited an affinity similar to that of melatonin itself, as well as full agonist activity. Optimization of the C-2 substituent by introducing Br, phenyl, or COOCH3 (2b-d) resulted in a significantly enhanced affinity (in the picomolar range) and improved agonist biological activity. Compounds lacking the methoxy group and bearing an N-alicyclic group (2h-j) behaved as partial agonists or antagonists. PMID- 9207942 TI - A new rational hypothesis for the pharmacophore of the active metabolite of leflunomide, a potent immunosuppressive drug. AB - Leflunomide is one of the most promising disease-modifying antirheumatic drug now in clinical trials for the treatment of rheumatoid arthritis. Metabolic studies have indicated that leflunomide is rapidly processed in vivo to an active metabolite, A771726 (2). To identify the chemical characteristics necessary for the immunosuppressive activity of 2, configurational and conformational studies were carried out on the latter and its inactive analogues (ethyl 3-hydroxy-2-((4 (trifluoromethyl)phenyl)carbamoyl)but-2-enoate, 3a, and 3-hydroxy-2-nitro-N-(4 (trifluoromethyl)phenyl)but-2-enamide, 3b). These studies suggested that the pharmacophore responsible for the immunosuppressive activity of 2 is a beta-keto amide with the enolic hydroxy group cis to the amidic moiety. To verify this hypothesis, a new class of immunosuppressive agents was designed and synthesized. Their testing in vitro and in vivo identified compounds which were more potent than both leflunomide and 2 and above all confirmed our hypothesis as to the key structural and chemical determinants for the immunosuppressive properties of 2 and our compounds. PMID- 9207943 TI - (2-Methyl-5-(methylsulfonyl)benzoyl)guanidine Na+/H+ antiporter inhibitors. AB - The inhibition of the Na+/H+ exchanger during cardiac ischemia and reperfusion has been shown to be beneficial for the preservation of the cellular integrity and functional performance. The aim of the present investigation was to come up with potent and selective benzoylguanidines as NHE inhibitors for their use as an adjunctive therapy in the treatment of acute myocardial infarction. During the course of our investigations it became clear that the substitution ortho to the acylguanidine was of crucial importance for the potency of the compounds. 4 Chloro- and 4-fluoro-2-methylbenzoic acids 6 and 7 were prepared using the directed ortho metalation technique with the carboxylic acid as the directing group. With the LDA/methyl iodide system the 2-methyl group could be extended to an ethyl group. 4-Alkyl groups were inserted by the palladium-catalyzed cross coupling reaction into the 4-bromo-2-methylbenzoic acid methyl ester (20). Starting with benzoic acids 6-19, the methylsulfonyl group was introduced by a sequence of standard reactions (sulfochlorination, reduction, and methylation). 4 Aryl derivatives 68-75 were synthesized by the palladium-catalyzed Suzuki reaction. A large number of nucleophilic displacement reactions in the 4-position were carried out with S-, O-, and N-nucleophiles as well as with the cyano and trifluoromethyl group. Using the ester method, acid chlorides, or Mukaiyama's procedure, the 5-(methylsulfonyl)benzoic acid derivatives were finally converted to the (5-(methylsulfonyl)benzoyl)guanidines 165-267 with excessive guanidine. In some cases nucleophilic substitutions with pyridinols and piperidine derivatives were carried out at the end of the reaction sequence with the 4-halo-N (diaminomethylene)-5-(methylsulfonyl)-benzamides. Variations in the 4-position were most reasonable, but the volume of the substituents was of crucial importance. Substitution in the 3- and particularly in the 6-position led to considerable worsening of the inhibitory effects of the Na+/H+ exchanger. The 2 methyl compounds, however, showed without exception higher in vitro activities than their respective demethyl counterparts as they are exemplified by the reference compounds 266 and 267, obviously caused by a conformational restriction of the acylguanidine chain. The development compound (2-methyl-5-(methylsulfonyl) 4-pyrrolobenzoyl)guanidine, methanesulfonate (246) is a NHE-1 subtype specific NHE inhibitor, being 27-fold more potent toward the NHE-1 than the NHE-2 isoform. 246 was found to act cardioprotectively not only when given before an experimentally induced ischemia, but also curatively after the onset of symptoms of acute myocardial infarction when given prior to the induction of reperfusion. PMID- 9207944 TI - Synthesis and in vitro evaluation of two progressive series of bifunctional polyhydroxybenzamide catechol-O-methyltransferase inhibitors. AB - Two progressive series of molecules with two polyhydroxybenzamide substructures were synthesized and tested as potential inhibitors of catechol-O methyltransferase (COMT). These compounds were designed for the purpose of enhanced enzyme binding with duplicated substructures separated by a linker section of various lengths. Our results show that potency and mode of inhibition observed with the "bifunctional" compounds were a reflection of their bifunctional nature. Furthermore, potency and mode of inhibition were dependent on the length of the linker section. Of the assayed compounds, the optimum linker was found to be diaminopropane. For example, N,N'-1,3-propanediylbis(3,4 dihydroxybenzamide) and N,N'-1,3-propanediylbis(3,4,5-trihydroxybenzamide) demonstrated strong inhibitory action against COMT, with apparent Ki values of 0.3 and 6.0 microM, respectively. PMID- 9207945 TI - Synthesis and antitumor properties of N-[2-(dimethylamino)ethyl]carboxamide derivatives of fused tetracyclic quinolines and quinoxalines: a new class of putative topoisomerase inhibitors. AB - A series of tetracyclic quinoline- and quinoxalinecarboxamides were prepared, and their cytotoxicities were evaluated in a series of murine human tumor cell lines. Most of the quinoline derivatives were prepared by an adaptation of the Pfitzinger synthesis, followed by thermal decarboxylation and coupling with N,N dimethylethylenediamine via a mixed anhydride method using isobutyl chloroformate. The quinoline analogues showed cytotoxicities broadly similar to those of the known tricyclic acridine-4-carboxamide mixed topoI/II inhibitor DACA, with thieno and indeno analogues being the most active. They showed little decrease in potencies against the Jurkat human leukemia topo II-resistant lines JLA and JLC, suggesting their cytotoxicity does not result primarily from inhibition of topo II. The quinoxaline analogues had more varied IC50 values, being on average less cytotoxic than the quinoline derivatives, but appeared to have a similar mode of action. Overall, this new class of compounds appear to be mixed topo I/II inhibitors, up to 3-fold more cytotoxic than DACA in the human leukemia cell lines studied, with in vivo activity in colon 38 comparable to that of DACA and doxorubicin. PMID- 9207946 TI - Structure-activity relationship of newly synthesized quinoline derivatives for reversal of multidrug resistance in cancer. AB - The effect of 24 newly synthesized quinoline derivatives on tumor cell multidrug resistance (MDR) was examined in vitro. At low concentrations, these compounds enhanced the accumulation of [3H]vincristine in K562/ADM cells and reversed tumor cell MDR. The results of the structure-activity relationship analysis indicate that in highly active compounds the two aryl rings in the hydrophobic moiety deviate from a common plane, so they are capable of interacting with hydrogen bond donors of P-170 glycoprotein (P-gp) via pi-hydrogen-pi interactions. Other major structural features which influence the MDR-reversing activities of these compounds are a quinoline nitrogen atom and a basic nitrogen atom in piperazine. Furthermore, in highly active compounds, the distance between the hydrophobic moiety and the basic nitrogen atom (an atom connected to 2 hydroxypropoxyquinoline) must be at least 5 A. Several compounds were found to reverse vincristine resistance in K562/ADM cells in vitro, and compound 16 (MS 209) was selected for clinical studies. PMID- 9207947 TI - 8-(1H-imidazol-1-yl)-7-nitro-4(5H)-imidazo[1,2-alpha]quinoxalinone and related compounds: synthesis and structure-activity relationships for the AMPA-type non NMDA receptor. AB - As a part of our program to discover novel antagonists for the AMPA subtype of EAA receptors, we designed and synthesized a series of heterocyclic-fused imidazolylquinoxalinones 5a-c, 9, 11, 14a-e, and 18 which led from 6-(1H-imidazol 1-yl)-7-nitro-2,3(1H,4H)-quinoxalinedione hydrochloride (1a.HCl, YM90K) by replacement of its amide with the imidazole and triazole rings. Their activity was evaluated by inhibiting [3H]AMPA binding from rat whole brain. As a result, it appeared that 8-(1H-imidazol-1-yl)-7-nitro-4(5H)-imidazo[1,2 alpha]quinoxalinone (5a) and its [1,2,4]triazolo[4,3-alpha] analogue 14a possessed high affinity for AMPA receptors with Ki values of 0.057 and 0.19 microM, respectively, similar to the activity of 1a and NBQX (2) (1a, Ki = 0.084 microM; 2, Ki = 0.060 microM). In contrast, 8-(1H-imidazol-1-yl)-7-nitro-4(5H) imidazo[1,5-alpha]quinoxalinone (5b) and 7-(1H-imidazol-1-yl)-8-nitro-4(5H) [1,2,4]triazolo[4,3-alpha]quinoxalino ne (18) showed no or weak affinity for the receptors. Hence, we deduced that the nitrogen atom of the fused heterocycles at the 3-position of 5a and 14a plays an essential role as hydrogen bond acceptors in binding to AMPA receptors, whereas their amides act as proton donors. From the SAR on 1-alkyl derivatives of 5a and 14a, it was indicated that introduction of suitable 1-alkyl substituents led to a severalfold improved AMPA affinity. A computational study on a model of water-quinoxaline complexes, a mimic of the putative hydrogen-bonding interaction between the receptors and quinoxalines, indicated that the different affinities of 5a, 14a, 1a, and 19 for the AMPA receptor may depend on, at least in part, each stabilization energy for the interaction. On this basis, we propose a pharmacophore model of AMPA receptors for the binding of the imidazolylquinoxaline derivatives. The heterocyclic-fused quinoxalinones 5a,c and 9 showed potent inhibitory activity in KA-induced toxicity for hippocampal cell culture with IC50 values of 0.30, 0.32, and 0.30 microM, respectively (1a, 0.81 microM; 2, 0.38 microM). Moreover 5a possesses over 5000-fold AMPA selectivity against both the NMDA receptor and the glycine site on the NMDA receptor. PMID- 9207949 TI - Non-peptide RGD surrogates which mimic a Gly-Asp beta-turn: potent antagonists of platelet glycoprotein IIb-IIIa. AB - Cyclic heptapeptide 1, which contains an Arg-Gly-Asp sequence, has good affinity for the platelet receptor GPIIb-IIIa and was chosen for study by 1H NMR techniques. The key RGD sequence of this molecule was found to reside in a conformationally defined type II' Gly-Asp beta-turn, and this information was used in the design of simple non-peptide RGD mimics. Disubstituted isoquinolones, bearing an acidic side chain at position 2 and a basic side chain at position 6, were prepared and were found to have modest affinity for GPIIb-IIIa. Systematic modification of the basic residue contained in these molecules yielded compounds with high affinity for GPIIb-IIIa. PMID- 9207948 TI - Discovery of an orally active series of isoxazoline glycoprotein IIb/IIIa antagonists. AB - Using isoxazoline XR299 (1a) as a starting point for the design of highly potent, long-duration GPIIb/IIIa antagonists, the effect of placing lipophilic substituents at positions alpha and beta to the carboxylate moiety was evaluated. Of the compounds studied, it was found that the n-butyl carbamate 24u exhibited superior potency and duration of ex vivo antiplatelet effects in dogs. Replacement of the benzamidin-4-yl moiety with alternative basic groups, elimination of the isoxazoline stereocenter, and reversal of the orientation of the isoxazoline ring resulted in lowered potency and/or duration of action. PMID- 9207950 TI - Structure-activity relationships of diverse Annonaceous acetogenins against multidrug resistant human mammary adenocarcinoma (MCF-7/Adr) cells. AB - Fourteen structurally diverse Annonaceous acetogenins, representing the three main classes of bis-adjacent, bis-nonadjacent, and single-THF ring(s), were tested for their ability to inhibit the growth of adriamycin resistant human mammary adenocarcinoma (MCF-7/Adr) cells. This cell line is resistant to treatment with adriamycin, vincristine, and vinblastine and is, thus, multidrug resistant (MDR). Among a series of bis-adjacent THF ring acetogenins, those with the stereochemistry of threo-trans-threo-trans-erythro (from C-15 to C-24) were the most potent with as much as 250 times the potency of adriamycin. A spacing of 13 carbons between the flanking hydroxyl of the THF ring system and the gamma unsaturated lactone seems to be optimum with a spacing of 11 and 9 carbons being significantly less active. Several single-THF ring compounds were also quite potent with gigantetrocin A (11) being the most potent compound tested. The acetogenins may, thus, have chemotherapeutic potential, especially with regard to MDR tumors. PMID- 9207951 TI - Radioiodinated estramustine phosphate and estramustine binding protein antibody accumulate in the prostate of a mouse. AB - BACKGROUND: The purpose of this study was to determine the distribution of radioiodinated estramustine (RI-EMP) and a radioiodinated antibody against estramustine binding protein (RI-EMBP-AB) in mice. METHODS: RI-EMP and RI-EMBP-AB were injected in male mice intravenously, and the activities of tissue samples were measured 1-31 hr from the injection. Pure iodine-125 (RI) was used as a control. RESULTS: RI-EMP accumulated in the prostate, which contained 2.6% of injected activity (ID) per gram tissue at 7 hr. The liver had an activity of 21.4% ID/g at 1 hr, which decreased as RI-EMP was secreted in bile. The lung contained 2.3% ID/g at 7 hr, and it retained the activity longer than the prostate. RI-EMBP-AB accumulated in the prostate: The activity was 2.9% ID/g at 7 hr. The gallbladder contained 6.5% ID/g at 7 hr. CONCLUSIONS: Due to its cytotoxic and radiosensitizing properties, RI-EMP can possibly be used for treating prostate cancer and other tumors. PMID- 9207952 TI - Expression of the apoptosis suppressing protein bcl-2 in prostatic adenocarcinoma is related to tumor malignancy. AB - BACKGROUND: The expression of bcl-2 protein has been related to histopathological features and prognosis in several epithelial tumors. In prostatic adenocarcinoma the prognostic significance of bcl-2 expression is mainly unexplored. METHODS: The expression of bcl-2 protein was assessed in 235 prostatic adenocarcinomas by using monoclonal bcl-2 protein antibody after microwave pretreatment of the cancer sections. The results of immunohistochemistry were related to histopathological features and prognosis of the patients. RESULTS: 71% of the tumors were bcl-2 negative, in 18% of cases the expression was weak, and in 11% of cases strong. The fraction of bcl-2 positive cells was variable (mean [SE]: 15 [2]%). The expression of bcl-2 protein was positively correlated to high T category, metastatic disease, and poor histological differentiation of the tumor. Tumors that were aneuploid and rapidly proliferating frequently expressed bcl-2 protein, while tumors we densely infiltrated by inflammatory cells rarely expressed bcl-2. The expression of bcl-2 protein was related to lowered survival probability in univariate analysis, while in multivariate analysis the expression of bcl-2 had no independent prognostic value. CONCLUSIONS: The expression of bcl 2 protein in prostatic adenocarcinoma is related to tumor malignancy, but the prognostic significance of the expression requires further analyses, particularly in localized tumors. PMID- 9207954 TI - Elastic system of the rat ventral prostate and its modifications following orchiectomy. AB - BACKGROUND: The extracellular matrix (ECM) has important roles in prostatic development, and marked stromal changes take place in the rat ventral prostate (VP) after androgen deprivation. However, little knowledge exists about individual ECM components. METHODS: The distribution of elastic fibers (EF) and elastic-related fibers (ERF) in the VP of castrated and control rats was investigated, using histochemistry and transmission electron microscopy (TEM). RESULTS: EF are barely detected in the prostatic stroma, but ERF are relatively abundant. Castration results in a relative increase in the number and thickness of ERF. TEM showed an open network of ECM microfibrils throughout, the stroma and thin and short EF, which increase in number and thickness after orchiectomy. CONCLUSIONS: The presence of elastic system components in the rat VP warrants the deformability required for the secretion exclusion under the action of smooth muscle cells, and the castration-induced modification may be related to the contraction of the tissue and maintenance of peculiar arrangements of other ECM components. PMID- 9207953 TI - Abnormal prostate development in C3(1)-bcl-2 transgenic mice. AB - BACKGROUND: Recent hypotheses to explain the etiology of abnormal growth associated with prostate disease have invoked perturbations in the rate of apoptosis as an important contributor to the onset and progression of these diseases. For this reason, the apoptosis suppressing oncoprotein bcl-2 has come under scrutiny with regards to its role in prostate diseases. In order to evaluate the role of bcl-2 in human prostate disease and to develop an animal model to test anti-bcl-2 therapies, we generated transgenic mice in which bcl-2 expression is targeted to the mouse prostate gland. METHODS: Mouse embryos were microinjected with recombinant DNA constructed by fusing a modified rat C3(1) promotor element to cDNA encoding human bcl-2. Presence of the C3(1)-bcl-2 transgene in progeny was identified by Southern blot and polymerase chain reaction (PCR) analysis. RNase protection assays were used to analyze RNA from 15 organs of these mice. Western blot assays and immunohistochemical staining were used to confirm the tissue-specific protein expression of human bcl-2 and its cellular localization. RESULTS: Three lines of C3(1)-bcl-2 transgenic mice were established. Founder mice carried 2-20 copies of the transgene. Expression of human bcl-2 from the transgene was limited to the prostate gland and testis of males as well as the uterus of females. In the prostate gland, human bcl-2 protein was found only in prostatic epithelial cells. Microscopic analysis of prostate glands from individual males (three lines) showed that these glands were often abnormal, with increased accumulation of cells in the prostatic stroma as well as the epithelium. CONCLUSIONS: These transgenic mice appear to provide a novel animal model for studying neoplastic development of the prostate, with particular emphasis on the bcl-2 protein and the role of apoptosis regulation in such development. PMID- 9207955 TI - Differentiation of rat neonatal ventral prostates grown in a serum-free organ culture system. AB - BACKGROUND: Organ culture methods have long been used in the study of the prostate because effects of drugs and hormones can be examined in the absence of systemic effects. METHODS: Neonatal rat ventral prostates (VP) were grown on Millipore filters floating on fluid medium composed of Dulbecco's modified Eagle's medium/Ham's F-12 supplemented with insulin, transferrin, and hydrocortisone, and in the presence or absence of testosterone (T, 10(-8)M). RESULTS: In the presence of T, ductal lumen formation occurred, ductal branching was extensive, and basal and luminal epithelial cells were identified by immunocytochemistry based on their distinctive cytokeratin profile. In the absence of T, ductal lumen formation did not occur, basal and luminal epithelial cells failed to differentiate, and there was a marked decrease in prostatic organ size relative to glands grown with T. Interestingly, DNA synthesis, as measured by counts per min (CPM) for 3H-thymidine incorporation, showed that DNA synthesis per microgram DNA at 7 days of organ culture was not inhibited by lack of T. Androgen receptor expression is another marker of prostatic epithelial differentiation, and it occurred in both the presence and absence of T. CONCLUSIONS: Growth and differentiation of the neonatal rat prostate in vitro occur in a manner similar to that of the developing prostate in vivo, demonstrating that organ cultures of neonatal rat ventral prostates provide a faithful model for studying rat prostatic development and differentiation under serum-free conditions. PMID- 9207956 TI - Rat prostate explants in serum-free organ culture: a comparison of two media and gas mixtures. AB - BACKGROUND: Urologists are looking for a way to easily discriminate between aggressive and very slow-growing prostate tumors. A sound way to appreciate such developing activities would be to identify an appropriate cell marker in prostate explants maintained in a defined culture system. METHODS: Different biological parameters were compared in rat prostate explants cultured for 5 days in rich CMRL or basic Leibovitz's L-15 medium, unsupplemented with serum, under a mixture of either 95% air/5% CO2 or 50% N2/45% O2/5% CO2. RESULTS: DNA synthesis was somewhat similar with the two-gas combination, but was higher in explants maintained in L-15 medium than in CMRL. Hence, L-15 medium and the 95% air/5% CO2 mixture were selected. Under these defined conditions for 5 days, cells were still able to synthesize DNA and proteins while preserving their morphological integrity and maintaining alkaline and acid phosphatase activities. CONCLUSIONS: Since the present culture system works well in a controlled environment and under such minimal conditions, it appears to be a reliable and promising model that will provide basic data and allow the study of hormones and growth factors involved in prostatic tissue growth. It might eventually permit the identification of a cell marker. PMID- 9207957 TI - Immunohistochemical analysis of estramustine binding protein with particular reference to proliferative activity in human prostatic carcinoma. AB - BACKGROUND: The estramustine binding protein (EMBP) specifically binds to estramustine and was first discovered in the rat ventral prostate. However, the physiological property of EMBP in the human prostate still remains to be elucidated. To elucidate whether EMBP is interrelated with cellular proliferation in human prostatic carcinoma (PC), the change in EMBP immunostaining during luteinizing hormone-releasing hormone (LH-RH) analog administration or during Cis platinum-based chemotherapy, and the difference in EMBP immunostaining between hormone refractory (hr-PC) and untreated PC were analyzed. METHODS: Forty-six patients with histologically proven untreated PCs (34 were treated with LH-RH analog and 12 were treated with chemotherapy as an initial therapy) and 14 with hr-PC were used in this study. PC tissues were obtained before and 3 months after the initial therapy. The changes in immunostainings for EMBP, proliferating cell nuclear antigen (PCNA), and nm23 protein were compared with the change in serum prostate-specific antigen (PSA) level and the histological response during the treatment. RESULTS: The increased EMBP expression was observed in tumors with high histological grade and high clinical stage as well as in hr-PC. In untreated PC, EMBP expression weakly correlated with PCNA or nm23 protein immunoreactivity. In PC receiving LH-RH analog, EMBP expression was significantly reduced after treatment, however, no significant changes were observed in PCNA or nm23 protein immunoreactivity. In addition, EMBP expression before the treatment significantly correlated with the serum PSA change, while PCNA expression and nm23 protein immunoreactivity did not. On the other hand, no significant relationship was observed between histological changes induced by the LH-RH analog and immunostainings for EMBP, PCNA, and nm23 protein before treatment. In PC patients receiving chemotherapy, immunostainings for EMBP, PCNA, and nm23 protein were not significantly changed during the treatment. EMBP immunoreactivity was significantly higher in hr-PC than in untreated PC with paralleled change of PCNA expression and nm23 protein immunoreactivity. CONCLUSIONS: These observations indicate that EMBP is androgen regulated in some PCs. However, EMBP expression is demonstrated even in hr-PC and is interrelated with cellular proliferation especially in hr-PC. PMID- 9207958 TI - Prostate-specific antigen-detected prostate cancer (stage T1c): an analysis of whole-mount prostatectomy specimens. AB - BACKGROUND: Clinical and pathological staging of prostate cancer has been, and remains, problematic. Since prostate-specific antigen (PSA)-detected tumors are often discerned during "screening," what are their significance? METHODS: We analyzed 67 consecutive patients with stage T1c prostate cancer undergoing radical prostatectomy at our institution from August 1, 1991-September 12, 1995, and who had whole-mount specimen processing. Diagnosis was determined in all cases by transrectal ultrasound-guided biopsy. RESULTS: The mean age of our patients was 63 years, and the mean PSA at time of diagnosis was 8.6 ng/ml (median, 7.2 ng/ml). There was organ-confined cancer in 31/67 (46%) patients; 17/67 (25%) had periprostatic fat infiltration, and of these 5(7%) had seminal vesicle involvement. Thirty-one of 67 (46%) had positive surgical margins. Twenty two (33%) had a Gleason sum of > or = 7 in the final pathological specimen. Insignificant tumors (dominant tumor volume < 0.20 cc) were found in only 4 cases. Smaller tumors were more likely to be found when the PSA was < 10 ng/ml. Multifocal disease was found in 64/67 (96%) prostate specimens. CONCLUSIONS: This study adds impetus to the growing realization that nonpalpable prostate cancer, detected because of elevated PSA, is rarely insignificant. Our findings add further emphasis to the fact that patients diagnosed by PSA elevation have, for the most part, significant cancer that should be treated aggressively. PMID- 9207959 TI - Genetic predisposition to prostate cancer: possible explanations for ethnic differences in risk. AB - BACKGROUND: It seems unlikely that the large ethnic differences in prostate cancer risk can be explained completely by ethnic differences in diet or other lifestyle characteristics. Instead, the differences may be due to ethnic variation in endogenous factors, such as androgen metabolism or inherited susceptibility. METHODS: We have reviewed the literature for evidence and support of ethnic variation in genetic susceptibility to prostate cancer as a reason for the ethnic differences in rates. RESULTS: We distinguish two types of ethnic variation: 1) variation in the prevalence of certain alleles of specific genes that confer modestly increased risk. Such variation might be reflected in ethnic differences in serum levels of androgens, their metabolites, or indicators of metabolism in the prostate; 2) variation in the prevalence of rare germline mutations conferring substantially increased risk. Such variation would be reflected in ethnic differences in familial aggregation of prostate cancer. We discuss the evidence in support of each of these two possibilities. CONCLUSIONS: Ethnic variation in polymorphic alleles of genes associated with modest fluctuations in risk could explain a large proportion of the ethnic difference in cancer risk. In contrast, rare mutations associated with substantially increased risk are likely to account for a smaller fraction of these differences. PMID- 9207960 TI - Induction of human cytotoxic T lymphocytes specific for prostate-specific antigen. PMID- 9207961 TI - [Homeoproteins and pituitary adenoma]. AB - Several transactivating factors specifically involved in the differentiation and proliferation of anterior pituitary cell types have been recently identified. Among them Pit-1 a member of the POU-domain transcription factors family is specific of anterior pituitary cells, and was initially identified and cloned as a transactivator of the GH and PRL genes and as a regulator of the TSH beta gene. Pit-1 play a key role during embryogenesis in the differentiation and proliferation of somatotrophs, lactotrophs and thyreotrophs. The importance of Pit-1 as a regulator in the anterior pituitary development has been further demonstrated by naturally occurring mutations or delections in dwarf mouse strains. In the Snell and Jackson dwarf mice, the levels of Pit-1 gene expression are low or undetectable, GH, PRL and TSH beta gene expression are absent and lactotrophs, somatotrophs, and threotrophs fail to proliferate. Furthermore Pit-1 carries out similar functions in humans. This is supported by the fact that children with mutations of the Pit-1 gene present with a congenital combined GH, PRL and TSH deficiency analogous to the phenotype of the Snell and Jackson dwarf mice. In children who were born to healthy consanguinous parents and present such combined deficiencies we recently reported a Pit-1 mutation causing a transition from a Phe to a Cys in a region of the protein known to be involved in DNA binding. Pit-1 transcripts identical in size and sequence to those observed in normal pituitary were described in human GH, PRL and TSH secreting pituitary adenomas. The Pit-1 beta isoform, raised through alternative splicing of exon 2 of the Pit-1 gene, is a more potent inducer of GH transcription than the major Pit-1 form. However no difference in the level of expression of the different Pit 1 isoforms was observed between tumors identified as pure GH or PRL producing tumors. The results support the existence of other transcription factors interacting with Pit-1 to coordinately regulate the activity of the GH and PRL promotors in a cell specific manner. In contrast, variable Pit-1 expression was observed in prolactinomas, according to their sensitivity to bromocriptine treatment. A highly significant correlation was indeed evidenced between the D2 receptors mRNA and the Pit-1 mRNA levels. These results raise the possibility that Pit-1 may either directly or indirectly affect the transcription of the D2 dopaminergic receptor gene. In fact, receptors for other hypothalamic neurohormones such a GHRH and somatostatin are known to be potential Pit-1 target genes. Such mechanisms could be implicated in the differentiation and proliferation of lactotrophs and somatotrophs. PMID- 9207962 TI - [Intra-sellar non-adenomatous expansive process]. AB - More than thirty types of tumors in the sellar region can mimic pituitary adenoma on, magnetic resonance imaging. When they exist, clinical manifestations are not necessarily highly contributive to diagnosis. Headache, visual impairment, signs of antepituitary insufficiency or possible dysmenorrhea with galactorrhea attributed to hyperprolactinemia due to compression of the dopaminergic axis are not specific and may be misleading. Clinical signs of diabetes insipidis and polyphagia are however suggestive of non-pituitary tumors. Consequently, high resolution imaging (MRI) and sometimes particular diagnostic circumstances (post partum for hypophysitis for example, or breast cancer for metastasis) orient the diagnosis. More rarely tumor enlargement, for example in certain germ cell tumors, provides a clue. PMID- 9207963 TI - [Pituitary radiotherapy. Current data and future prospects]. AB - Some technical improvements have allowed to minimize the frequency of severe complications following fractionated pituitary conventional radiotherapy, without altering its efficiency. "Conformational" radiotherapy is currently under development, aiming at the best fitting of the tumor borders to the irradiation zone, by the means of stereotactic imaging. More recently, radiosurgery has been proposed for pituitary adenomas. It consists in a single high radiation dose to the tumor, by the means of either cobalt minibeams (Gamma Unit) or photon beams from a linear particle accelerator. These techniques require the use of a stereotactic frame and precise 3D imaging in order to tightly superimpose the target volume to the reference isodose. They must not be viewed as an alternative to conventional radiotherapy. They can be applied only to small lesions (less than 20 mm in their maximal axis) which are distant (> 5 mm) from the optic chiasma and nerves. Their efficiency is similar to the one of fractionated conventional radiotherapy, with a shorter response time. In conclusion, radiotherapy can be used safely for pituitary adenomas. It remains however a second line treatment, when surgery has been incomplete and when a simple, effective and inexpensive medical treatment is not possible. PMID- 9207964 TI - [Paracrine regulation of anterior pituitary hormones by neuropeptides]. AB - The hypothalamus is the source of neuropeptides which, being secreted into the portal system, control the synthesis and the secretion of the anterior pituitary hormones. Besides the well characterized hypothalamic central control and the hormonal peripheral control, recent studies have shown, in the anterior pituitary, the expression, among many other regulatory factors, of neuropeptides that are identical to those produced by the hypothalamus and that seem involved in the local control of anterior pituitary functions through autocrine or paracrine mechanisms. The presence of the neuropeptide mRNAs, precursors and mature forms of the peptides in anterior pituitary tissues as well as the secretion of the mature peptides argue in favor of the intrinsic ability of the normal and tumoral anterior pituitary to express neuropeptides. This expression of neuropeptides occurring in tissues bearing functional receptors for these ligands, anterior pituitary control could rely, at least in part, on endogenous neuropeptides acting locally. Correlations between neuropeptide contents in the anterior pituitary and the plasma levels of anterior pituitary hormones suggest that neuropeptides of anterior pituitary origin can play a local regulatory role, complementary of the classical hypothalamic central control. In the normal anterior pituitary which remains under hypothalamic control, it is presently difficult to evaluate the relative importance of the local and central control. However, anterior pituitary hyperplasia and pituitary tumors represent two models in which the specific contribution of the local control is easier to define. PMID- 9207965 TI - [Peroperative detection probes. Evaluation and perspectives in endocrinology]. AB - The principal primary endocrine tumors may be visualized scintigraphically by injection of highly specific radiopharmaceuticals. When surgery is indicated, the surgeon may be assisted in difficult cases by a probe detecting the radioactivity concentrated in tumoral tissue. A radiopharmaceutical consists of a vector molecule labeled with a radioactive isotope. Only radioactive emissions with a certain half life and energy are suitable for intraoperative detection; this limits the number of usable radionuclides. In practice, one uses iodine 123 (123I), iodine 125 (125I), and iodine 131 (131I) labeled metaiodobenzylguanidine mIBG or indium 111 (111In) labeled pentetreotide (Octreoscan) or bi-specific anti CEA antibody. During intra-operative detection, the probe is closely positioned to the tumor-tissue which is surrounded by radioactive background activity due to the non-specific binding in adjacent organs. In order to work with probes that are easily handled, and sensitive at the same time, one can opt for technology which uses either diodes or scintillating crystals. These probes can be used at room temperature and their choice depends upon the energy to be detected. Interesting results begin to be published concerning pheocyromocytomas, differentiated thyroid carcinomas, gastro-entero-pancreatic tumors and medullary thyroid carcinoma, especially for reintervention when conventional imaging modalities fail to localize tumors. PMID- 9207966 TI - [Value of myocardial exploration by isotopic technique in diabetes]. PMID- 9207968 TI - [Is celioscopic approach of pheochromocytoma acceptable? Reflections apropos of a prospective study of 6 personal cases]. AB - Today, laparoscopy is for us the technique of choice for approaching presumed benign adrenal tumors. With regards to pheochromocytoma however, two major questions must be addressed. First, is it acceptable to resect potentially multifocal tumors with such a targeted approach? Second, can peroperative hemodynamic changes be anticipated and controlled by the anesthetist, taking into account the additional effects of pneumoperitoneum and catecholamine release on the cardiovascular system? The present prospective study attempts to answer these two questions. From November 1993 to November 1995 we operated on four women and two men, with ages ranging from 33 to 71 years (mean of 47) and a mean Body Mass Index of 25 kg/m2 (range 17-35). Four patients were assigned ASA (American Society of Anesthesiologists) physical status 2, one grade 1 and one grade 3. Comprehensive preoperative work-up, including a CT scan and an I131 MIBG Scan in all, a C11 Hydroxyephedrine PET Scan in 4 and a MRI in one patient, showed a solitary lesion in each case. There were four right-sided and two left-sided tumors, ranging from 30 to 60 mm in diameter. Laparoscopy was always performed transperitoneally. Systemic and pulmonary hemodynamics were thoroughly assessed. Epinephrin and norepinephrin concentrations were measured at the 10 key-time of surgery. Use of continuous intravenous infusion of nicardipine allowed tight control of hemodynamics despite impressive increases in circulating catecholamines. The mean operative time was 76 minutes (range 59-130). Blood loss was minimal. We observed neither mortality nor morbidity. Mean hospital stay ranged from 3 to 13 days (median = 3). All patients are normotensive without drug after a follow-up of 9 to 33 months. In conclusion, we think that laparoscopic removal of selected cases of pheochromocytoma may be performed safely from both the hemodynamical and oncological standpoints. PMID- 9207967 TI - [Value of scintigraphic explorations by radiomarkers others than iodine radioisotope in differentiated thyroid cancer]. AB - Radioiodine scintigraphy is the gold standard exploration for imaging metastases of differentiated thyroid cancer and enables the decision of therapy with 131 radioactive iodine to be made. However, other approaches may be of use for diagnosis when there is no visible uptake after the administration of 131I, while elevated thyroblobulin levels suggest the presence of metastatic tissue in one third of metastatic patients. In order to detect recurrences or metastases, in conjunction with conventional imaging techniques (cervical and hepatic ultrasonography, lung CT scan..), other scintigraphic explorations with various radiopharmaceutics may be used, although none of them has any specificity towards thyroid cancer. Tl201 and MIBI which are used as perfusion tracers for myocardial explorations, are also used for detection of various tumors and for metastatic thyroid cancer. The performances of both radiopharmaceutics in imaging metastases are differently evaluated between investigators with a sensitivity ranging from 45 to 94% while the specificity varies less (82-97%). 18-Fluoro-deoxyglucose is retained in malignant tissue depending on the grade of malignancy. It has been shown to accumulate in thyroid cancer and metastases. Its detection by whole body PETscan represents a limitation for use which will be modified by new techniques. 111In-octreotide which binds to somatostatin receptors located on tumor cell membranes is able to show thyroid cancer metastases in some instances. We report on the very preliminary results of these combined scintigraphic approaches, performed in a limited number of patients who had no radioiodine uptake and elevated Tg levels, in order to determine the most appropriate exploration in terms of performance and cost. PMID- 9207969 TI - [Malformations of the nasal fossa and paramedian facial clefts. New perspectives]. AB - Since choanal atresia may be associated with other cranio-facial malformations, including various degrees of nasal fossa malformation, and be a part of paramedian facial clefts, (as described by Tessier), they can be integrated into the larger group of neurocristopathies. We identified four such cases with combined clinical elements corresponding to Tessier's paramedian facial cleft, including eyelid coloboma, mild to severe choanal and nasal fossa anomalies, ethmoidal hypoplasia and anterior skull base malformation, sometimes with proboscis lateralis. These various malformations are due to abnormality of the olfactive placode and the adjacent mesenchyme. These discoveries incited us to elaborate a conception first of all on the pertinent embryology involved, second, to propose a new classification based on anatomical and pathogenic embryological considerations. And finally, since endonasal laser therapy is particularly dangerous in such cases, to propose the use of transpalatal approach to restore choanal permeability. Pediatric ENT surgeons should pay special attention to any small stigmatism of facial cleft when dealing with children affected by choanal atresia. PMID- 9207971 TI - [Frontal mucoceles of orbital or cerebral extension: therapeutic strategy]. AB - The aim of this study was to investigate ten patients who underwent surgery for mucoceles of the frontal sinus. Bicoronal direct access was required by the localization of the mucocele limited to the frontal sinuses, its extension to the orbit and/or the brain and because of the anatomy of the frontal sinuses (large size, lateral horn...) as evidenced at imaging. Direct access to the frontal sinuses was achieved in 9 patients allowing marsupialization associated with repermeabilization of the naso-frontal duct (7 cases) or exeresis of the mucocele by cranialization (2 cases). Mean follow-up is 27 months. Repermeabilization of the naso-frontal duct was effective in 7 out of 8 cases. There were no complications after cranialization and no recurrence has been observed. Drainage of frontal mucoceles is a first intention strategy. In case of complication or recurrence, cranialization of the frontal sinuses would appear to be better than an exclusion-filling procedure. PMID- 9207970 TI - [Efficacy of partial inferior turbinectomy in the treatment of nasal obstruction. Retrospective study apropos of 71 patients]. AB - Longtime discredited, practiced with reticence even today, the inferior turbinectomy represents the treatment of choice in patients with persistent chronic nasal obstruction despite medical treatment or cauterization. Between January 1989 and December 1995, 81 patients underwent an isolated bilateral inferior turbinectomy (without an associated septal or sinus intervention). This retrospective study analyzes the short and long-term results in 71 patients with a one year minimum follow-up. Turbinectomy was in all cases always partial and managed under endoscopic guiding. Mean follow-up was 33 months, based on a patient's questionnaire. No cases of crusting rhinitis were observed. 81,7% of patients were improved by the surgical intervention. In asthmatic patients, no worsening of the asthma was noted, whereas in 28,5% of these cases, there was an improvement or disappearance. Non serious adverse effects appear after surgery as rhinorrhea (16%) and post nasal drip (18,4%). The surgical turbinate reduction represents an option in the care of patients with chronic nasal obstruction associated with inferior turbinate hypertrophy. PMID- 9207972 TI - [Unilateral choanal atresia and paranasal sinus growth]. AB - Mechanisms regulating sinus growth are poorly understood. We report a series of six cases of unilateral choanal atresia and discuss the role of nasal ventilation on sinus growth. The presence and the size of the sinus cavities are the main parameters. Our preliminary results suggest that sinus growth is independent of nasal ventilation. PMID- 9207973 TI - [Orbito-maxillo-facial rehabilitation with osteointegrated prostheses]. AB - This investigation was conducted to evaluate 20 cases of implant-retained prosthetic rehabilitation (mean follow-up of 35 months). Ear, nose, orbit and complex defects of the face have been restored. Our main indication was reconstruction in cancer patients after tumor surgery (n = 14). Rehabilitation was also performed in traumatic disorders (n = 3) and congenital aural malformations (n = 3). Fifty seven titanium fixtures have been implanted with an integration success rate of 98%. We also report 6 cases of maxillary or mandibulary functional rehabilitation. The use of osteointegrated implants to provide support for maxillo-facial prosthesis is a new and valuable technique when reconstructive plastic surgery is not envisageable. PMID- 9207975 TI - [Horizontal subglottic laryngectomy in treating cancers of the laryngeal vestibule and vallecula. Apropos of 65 cases]. AB - A retrospective review of 65 patients treated with supraglottic laryngectomy from 1980 to 1992 in the Department of Otolaryngology of Rennes was performed. Tumor localisation was vallecula in 38 cases and epiglottis in 27 cases. Functional and carcinologic results are studied for all patients, when base tongue is involved and for patients operated after radiotherapy. The 3 years survival rate was 62,2% and 5 years survival rate 44%. A local failure occurred in 15,4% and 4,6% developed cervical metastasis. Another neoplasm on superior aerodigestive tract is the first cause of death for epiglottis cancer. This data shows that conservative surgery is a higher effective method than radiotherapy for the treatment of small tumors of epiglottis. PMID- 9207974 TI - [Crista ampullaris mechanics and possibilities of regulation of the semi-circular canal]. AB - A software using a finite element method is used to study the deformation of a bidimensional model of the human cupular diaphragm and the distribution of angular shearing of the hairs of the sensory cells located on the crista ampularis. This study shows that the hairs on the top of the crista are more sensitive to the pressure difference across the cupula due to angular acceleration of the head and those located on the sides of the crista are sensitive to the variation of the inflation pressure of the ampulla. This spatial discrimination of sensibility of the hair cells to the two types of mechanical solicitations leads us to consider the SCC as regulated sensor. PMID- 9207976 TI - [Malignant tumors of the parotid gland. Apropos of 60 cases]. AB - Among a series of 520 parotidectomies performed between 1975 and 1995, we observed 88 cases of malignancy (17%). For this study, we excluded skin cancers which had invaded the parotid and glandular metastases of squamous cell carcinoma, malignant melanoma or kidney cancers, retaining only tumors with a glandular origin and lymphomas. Thus defined, our series comprised 60 patients, i.e. 12% of the operated parodids (31 confirmed cancers, 18 tumors with intermediary malignancy, including several in which the pathology report confirmed malignancy, and 11 lymphomas). We examined therapeutic management by histology and compared the outcomes. Relation with the facial nerve are discussed. Prognosis depends on histology, tumor stage and treatment. PMID- 9207977 TI - [Prospective study of thromboses of the subclavian vein after setting implantable venous access systems in cervico-facial cancerology]. AB - Seventy-eight consecutive patients treated by chemotherapy for ENT cancers and having a subclavian catheter for venous access were studied prospectively to assess the prevalence of venous thrombosis. Thrombosis of the subclavian vein was demonstrated clinically in 4 patients and by ultrasonography in 7 patients. The prevalence of thrombosis was 14.1%. No clinical or biological predisposition factor could be identified. Subclavian thrombosis mostly occurred during the second month after implantation (91% of cases). Ultrasonography seems the most useful non-invasive technique for the diagnosis of subclavian thrombosis. PMID- 9207978 TI - [Sudden deafness and Barlow disease]. AB - From January 1993 to December 1994 twelve patients were evaluated for sudden hearing loss. The median age was 49 years with a range of 18 to 71. All had severe or profound initial hearing loss. The incidence of bilateral disease was 25%. Total deafness occurred in five (33%) ears. Nine (75%) patients had vestibular symptoms and eight (67%) admitted experiencing tinnitus. Two dimensional echocardiography revealed mitral prolapse in eight (67%) patients; another patient showed moderate to severe ischemic left ventricular dysfunction with apical aneurysm. PMID- 9207979 TI - [Arteriovenous malformation of the mandible. Apropos of a case in a 6-year-old child]. AB - Arteriovenous malformations of the mandible are uncommon. Though characteristic clinical and radiological presentations are well described, management in the case of children is still a matter of debate. We report a mandibular angioma in a 6-year-old girl which was treated with superselective embolization. This method is the treatment of choice for mandibular arteriovenous malformations in children because of the low complication rate and the preservation of healthy tissue. When the malformation continues to develop despite repeated embolizations, surgical treatment may be necessary. PMID- 9207980 TI - Helicobacter pylori infection and genetic polymorphisms for cancer-related genes in gastric carcinogenesis. AB - The development of gastric cancer is a multistep process that is multi-factorial. An association with the Helicobacter pylori infections, gastric atrophy and gastric cancer has received recent attention. The objective of this study was to elucidate the risk factors for gastric cancer by using molecular epidemiological techniques for genetic susceptibility, gastric atrophy and serum markers including H pylori infection. We used an age- and gender-matched case-control study, where patients with benign gastric lesions were the controls. Low serum pepsinogen I levels (cut-off < 50 ng/mL) and low pepsinogen I/pepsinogen II ratios (cut-off < 3.0) were significantly associated with the risk of gastric cancer (odds ratio [OR] = 3.53: 95% confidence interval [CI] = 2.46-5.09 and OR = 4.73: 3.26-6.88, respectively). However, seropositivity of serum immunoglobulin G (IgG) antibody against H pylori (OR = 1.09: 0.74-1.61) was not associated with gastric cancer, even when analyzed by age greater than or less then 50 years. Specific genotypes of the cytochrome p450 2E1 (CYP2E1) RsaI polymorphism and glutathione-S-transferase (GST) M1 gene deletion were determined but were not associated with gastric cancer; however, a Lmyc genetic polymorphism was associated with gastric cancer (OR = 1.55: 1.03-2.34). Therefore, in this Japanese study, atrophic mucosal change, indicated by serum pepsinogen levels, is a possible risk factor for gastric cancer. PMID- 9207981 TI - Helicobacter pylori and atrophic gastritis. AB - Infection with Helicobacter pylori plays a crucial role in the etiology of atrophic gastritis and gastric cancer. Studies suggest a nine-fold increased risk for both conditions in the presence of infection. The risk of atrophic gastritis in the presence of infection is dependent upon the severity of the gastritis. Gastritis is increased in subjects infected with a cytotoxic H pylori strain and in those with a decreased acid production. The development of atrophy may be related to the induction of cross-reacting antibodies recognizing Lewis epitopes on H pylori lipopolysaccharide and gastric mucosa. Future studies have to demonstrate whether atrophic gastritis and gastric cancer can be prevented by early H pylori eradication. PMID- 9207982 TI - Eradication of Helicobacter pylori and regression of B-cell lymphoma. AB - There is an increasing body of evidence implicating a causal association between Helicobacter pylori and the development of mucosa-associated lymphoid tissue (MALT) associated B-cell gastric lymphoma. Investigators have noted that almost all patients with H pylori-associated chronic gastritis develop lymphoid follicles. Some of these patients demonstrate infiltration of B cells and lymphoepithelial lesions typical of MALT lymphoma. When gastric tissue from patients with gastric B-cell lymphoma is analyzed for H pylori infection, the overwhelming majority of patients demonstrate this condition. Epidemiologic nested case-control studies have shown that patients with gastric non-Hodgkin's lymphoma are substantially more likely that matched controls to have H pylori infection. This situation may be analogous to the linkage between chronic Epstein Barr virus and lymphoma. The mechanisms inducing the development of lymphoma are not clear, but it has been suggested that chronic infection with H pylori results in the stimulation of H pylori-responsive T cells which in turn activate B cells with the subsequent development of a mutation to a monoclonal B-cell population. Recent evidence indicates that cure of H pylori infection produces regression of MALT lymphoma within 3 to 12 months in approximately 75% of antibiotic-treated patients. Individual responsiveness remains unpredictable, however, and careful and prolonged endoscopic and histologic follow-up is needed. Large, well controlled studies are necessary, however, to determine the duration of 'cure' and the appropriate setting for treatment. PMID- 9207983 TI - The microbiology of Helicobacter pylori. AB - Helicobacter pylori was first isolated only in 1983, but has rapidly gained an extraordinary prominence in gastroenterology and medical microbiology. It lives only in the human stomach beneath the mucin layer which protects it from the lethal effects of gastric acid. Although it has a very high prevalence (30-40% in developed and more than 80% in developing countries), the route of infection is not known; gastro-oral and faecal routes have been postulated. The organism is associated with gastritis, duodenal and gastric ulcers and gastric cancer as well as a number of diseases at distant sites. It can be eradicated with combinations of antibiotics, after which the rate of infection is low. PMID- 9207984 TI - Fibrinolytic response in subjects with hypertriglyceridemia and low HDL cholesterol. AB - The combination of hypertriglyceridemia and low high-density lipoprotein cholesterol (HDL-C) appears to be an excessively high risk factor for coronary artery disease (CAD). In the Helsinki study, both coronary events and mortality were decreased by gemfibrozil, especially in subjects with low HDL-C and high triglycerides (TG). On the other hand, it is known that high levels of TG can be associated with high levels of circulating plasminogen activator inhibitor (PAI), which is also a possible risk factor for CAD. The aim of the present study was to see: 1) whether the combination of low HDL-C and high TG is associated with a more impaired fibrinolytic response than in either isolated condition, and 2) whether gemfibrozil administration can improve fibrinolysis in patients with both high TG and low HDL-C. Twelve non-obese, non-diabetic subjects (eight men, four women; mean age 55 +/- 13 yrs) with low HDL-C (< 35 mg/dL men; < 45 mg/dL women) and high TG (mean 253.6 +/- 42.6 mg/dL) entered the study (Group A). Additionally fourteen comparable subjects with normal HDL-C were also investigated (Group B), plus 12 comparable subjects with isolated low HDL-C (Group C). Ten healthy people served as the control group. The following plasma fibrinolytic parameters were measured: tissue plasminogen activator antigen, PAI antigen and activity, euglobulin fibrinolytic activity (EFA) on fibrin plates, plasminogen and alpha-2 antiplasmin activities. All except the latter two values were also measured after venous occlusion (vo). In baseline conditions, patients in Groups A and B had higher EFA values before vo and higher PAI-1 antigen and alpha-2-antiplasmin levels after vo than those of controls or the subjects in Group C. The relationship between PAI antigen and PAI activity and TG was not confirmed in our population (n = 48). We also saw no interference due to HDL-C, while there was a significant relationship between EFA before vo and both TG and cholesterol. After gemfibrozil treatment (600 mg bid for 12 weeks), the lipid profiles of subjects with high TG and low HDL-C were significantly improved. There was also a slight reduction of PAI activity after vo, while the PAI-1 antigen had decreased significantly from baseline after vo (56.3 +/- 13 ng/mL before vo; 48.4 +/- 21 ng/mL after vo; P = 0.04). The higher risk of CAD in patients with low HDL-C and high TG might be in part related to impairment of fibrinolysis, which occurs in patients with isolated high TG. The close relationship existing between both TG and cholesterol levels and fibrinolytic activity confirm the key role of this latter process in the development of CAD. PMID- 9207985 TI - Pharmacokinetics and hepatotoxicity of diclofenac using an isolated perfused rat liver. AB - Pharmacokinetics and hepatotoxicity of diclofenac was studied in a recirculating model of isolated perfused rat liver. Ten male Sprague-Dawley rat (weighing 230 330 g) livers were perfused for 2 h with 250 mL Krebs-Henseleit bicarbonate buffer that contained 10.75 mg (group A, n = 5) and 1.075 mg (group B, n = 5) of diclofenac (approximately 100 and 10 times the therapeutic dose in man, respectively). Samples were collected from the efflux at regular time intervals for the determination of diclofenac concentrations by a high performance liquid chromatography (HPLC) method. Pharmacokinetic analyses were carried out using a computer program. To establish viability of the liver and toxicity of the drug, enzyme activity measurements of lactate dehydrogenase (LDH), aspartate aminotransferase (SGOT) and piruvate aminotransferase (SGPT) were performed by a spectrophotometric method. Oxygen consumption was also recorded during the entire perfusion period. Both groups presented bicompartmental kinetics. Concentration profiles showed that group B had a better metabolizing capacity, reflected in a 85.54 +/- 37.05 min half-life, a 0.52 +/- 0.19 mL min-1 g-1 liver clearance and a 0.517 +/- 0.188 extraction ratio, compared to group A, which presented a 123.95 +/- 88.13 min half-life, a 0.1164 +/- 0.067 mL min-1 g-1 liver clearance (P < 0.002) and a 0.116 +/- 0.680 extraction ratio (P < 0.002). LDH activity showed a significant increase in group A at 90 min in comparison with the control group, while in group B this increase was significantly higher at 10 min (P < 0.004). The aminotransferase levels did not show a significant increase. According to these results, diclofenac would not have a direct hepatotoxic effect, even at doses 100 times higher than therapeutic ones. PMID- 9207986 TI - Anti-HIV activity of extracts from Calendula officinalis flowers. AB - Extracts of dried flowers from Calendula officinalis were examined for their ability to inhibit the human immunodeficiency virus type 1 (HIV-1) replication. Both organic and aqueous extracts were relatively nontoxic to human lymphocytic Molt-4 cells, but only the organic one exhibited potent anti-HIV activity in an in vitro MTT/tetrazolium-based assay. In addition, in the presence of the organic extract (500 micrograms/mL), the uninfected Molt-4 cells were completely protected for up to 24 h from fusion and subsequent death, caused by cocultivation with persistently infected U-937/HIV-1 cells. It was also found that the organic extract from Calendula officinalis flowers caused a significant dose- and time-dependent reduction of HIV-1 reverse transcription (RT) activity. An 85% RT inhibition was achieved after a 30 min treatment of partially purified enzyme in a cell-free system. These results suggested that organic extract of flowers from Calendula officinalis possesses anti-HIV properties of therapeutic interest. PMID- 9207987 TI - Enhanced functional capacity of the peritoneal macrophages as antigen presenting cells after aclacinomycin treatment in mice. AB - Aclacinomycin (ACM) is an oncostatic of the anthracycline family, largely used in patients and experimentally in mice. ACM has been reported to enhance phagocytosis, secretion of free oxygen radicals and of interleukin 1. Its injection is also followed by an increase of the cytotoxic and cytostatic activity of murine peritoneal macrophages. In the present work we investigated whether ACM modifies the antigen-presenting cell capacity of murine peritoneal macrophages. Purified T lymphocytes were cultured with peritoneal macrophages from either normal or ACM treated mice (4 mg/kg day -4) which were previously incubated with phytohemagglutinin. The T cell proliferative response was greater in cultures with normal macrophages, indicating that macrophages from ACM-treated mice had a better antigen presenting activity than normal untreated macrophages. PMID- 9207988 TI - [Information on the disabled and dependent people in France]. AB - For the time being information on the exact number of blind persons living in France is not available. A great deal of reasons contribute to such a surprising lack of knowledge. As a matter of fact the various attempts for defining the state of inability and/or dependency of a person are not fully promising. Moreover the level of inability- and dependency-is changing over time. During the next two or three decades, as the baby-boomers reach the old ages, the number of people of 65 years old and over will steadily be rising. There are some signs to predict that the number of unable and dependent people will grow at a lower pace. However, the industrialized societies should be prepared to the new challenge. As a first step, they should obtain a better knowledge regarding their dependent people. PMID- 9207989 TI - [Flourishing of micro-units of health care in Benin]. AB - The end of the marxist-leninist regime in Benin in 1990 has allowed a steady growth of what can be named "the micro-units of health care" at the periphery of cities. Demand for care from the urban poor and urban lower-middle classes is mainly directed towards these micro-units instead of the public (or private) official health centers. "Micro-units of health care" are small for-profit houses where primary care is delivered to buying patients by young health graduates who could not find a post in the official health system. Micro-units are blossoming in Benin, they are not registered and of course not controlled. The basic reasons for their success can be identified as followed: (i) modest investment, low cost and consequently low price; (ii) near the users; (iii) unsophisticated health care; (iv) no competition from the official health system; (v) the users give to these units a higher confidence than to the official health structures, the latter having of course a much higher medical standard; (vi) warm welcome and personalized services. It is probable that, twenty years ago, those who, in Alma Ata made the famous declaration on primary health care did not thought about these micro-units in Benin. It is certain that these micro-units are implementing a great deal of the principles highlighted by the Declaration of Alma Ata. PMID- 9207990 TI - [Vascular involvement in chronic rejection]. PMID- 9207991 TI - [Comparison of scanners and porto-scanners, both in helical mode, for the study of hepatic metastases. Prospective study in 12 patients]. AB - PURPOSE: The aim of this study was to compare helical CTAP and helical CT-scan in the preoperative assessment of liver metastases. METHODS: A prospective unicentric study in 12 patients was performed with helical CTAP and helical CT scan. All patients underwent partial hepatectomy with intraoperative palpation and sonography within 19 days (mean: 9 days). RESULTS: Examination of resected liver specimens found 38 metastases, from colorectal cancer in 36 cases. The sensitivity was 92.1% for helical CTAP and 79% for helical CT-scan. This sensitivity was 85% for helical CTAP and 60% for helical CT-scan for nodules 1 cm or less in diameter (P = 0.08). CONCLUSION: In the preoperative screening of liver metastases, helical CT-scan should be performed as the first choice examination. When hepatic lesions seem to be curable by resection based on helical dynamic CT-scan results, helical CTAP should be performed to increase the sensitivity of detection of lesions 1 cm or less in diameter. PMID- 9207993 TI - [Membranous obstruction of the inferior vena cava]. PMID- 9207992 TI - [Obliterative arteriopathy in chronic rejection after hepatic transplantation. Search of cytomegalovirus genome by in situ hybridization and viral antigens by immunohistochemistry]. AB - OBJECTIVES: This study was performed to assess the role of cytomegalovirus in the parietal infection of intrahepatic arteries in the pathogenesis of obliterative arteriopathy from chronic rejection after orthotopic liver transplantation. METHODS: We studied two groups of liver transplants by in situ hybridization and immunohistochemistry: group 1, including 10 liver grafts with obliterating arteriopathy, and group 2 including 7 liver grafts without any arterial disorders. The results were correlated with clinical data (cytomegalovirus infection and acute rejection). RESULTS: By in situ hybridization, cytomegalovirus DNA was identified in the media in 70% of transplants in group 1 and 42% in group 2. Detection of immediate early and late antigens by immunohistochemistry was negative. Cytomegalovirus infections were often associated with acute rejection. CONCLUSION: These results suggest that cytomegalovirus detected in arteries is latent, and that cytomegalovirus probably does not play a role in the pathogenesis of chronic rejection obliterative arteriopathy. PMID- 9207994 TI - [Intraductal papillary mucinous tumors of the pancreas. An emerging entity?]. PMID- 9207995 TI - [Intraductal papillary mucinous tumors of the pancreas. Clinical and morphological aspects in 30 patients]. AB - AIM: Intraductal papillary-mucinous tumors of the pancreas are rare and characterized by a malignant potential. The aim of this study was to clarify their clinical presentation and the performance of different imaging procedures to determine malignancy and tumor extent. METHODS: Medical records and radiographs of 30 patients with histologically confirmed intraductal papillary mucinous tumor of the pancreas were reviewed retrospectively. Imaging procedures were compared with pathological data of resected pancreas to evaluate their performances. RESULTS: The most frequent symptom was acute pancreatitis (37%). The onset of symptoms preceded the diagnosis by 2.5 years. Diabetes mellitus and diarrhea were respectively detected in 33 and 23% of the cases. The combination of CT scan, endoscopic retrograde cholangiopancreatography and endosonography allowed correct diagnosis of intraductal papillary-mucinous tumor of the pancreas in 100% of the cases. Tumor extent could be accurately determined considering the location of cystic dilatation of the pancreatic ducts, the presence of intraductal material or parietal irregularity. Actuarial 2-year survival rate was 43% in patients with malignant tumors. Radiological factors predicting malignancy were: vascular invasion, common bile duct dilatation, stricture of the main pancreatic duct and the presence of solid component in the tumor. CONCLUSION: The combination of CT scan, ERCP and endosonography provide accurate diagnosis of intraductal papillary-mucinous tumor of the pancreas as well as assessment of tumor extent and malignancy. PMID- 9207996 TI - [Recurrence after surgical treatment of epidermoid cancers of the esophagus. Therapeutic approach in 19 patients]. AB - OBJECTIVES: The aim of this study was to report the management of 19 patients with recurrence of esophageal squamous cell carcinoma after surgical treatment. PATIENTS-METHODS: Nineteen patients with loco-regional recurrent invasion (n = 13) or metastasis (n = 6) of esophageal squamous cell carcinoma were included. Four of the 13 patients with loco-regional recurrent invasion had tracheal involvement. The treatment of the recurrence was a combined radiochemotherapy (n = 12) for loco-regional recurrent invasion in 11 cases and for metastasis in 1 case, associated with a tracheal prosthesis in 1 patient. The other treatments were chemotherapy alone (n = 5), esophageal prosthesis (n = 1) and surgical treatment (n = 1). RESULTS: There were 7 objective responses among the 12 patients treated with combined radiochemotherapy and none in the group treated with chemotherapy alone. Grade 3-4 toxicity was noticed in 2 cases (severe mucositis). Survival rate of the 19 patients was 52.6% at 1 year and 13.1% at 2 years; it was linked with general health (P = 0.09) and with tracheal involvement (P = 0.04). Survival rate of the 12 patients treated by combined radiochemotherapy was higher: 66% at 1 year and 22.2% at 2 years (median survival time = 16 months). CONCLUSION: Active medical treatment of recurrence of esophageal squamous cell carcinoma by combined radiochemotherapy can provide a median survival time of 16 months, with a moderate toxicity. PMID- 9207997 TI - Effects of intrajejunal perfusion and chronic ingestion of Lactobacillus johnsonii strain La1 on serum concentrations and jejunal secretions of immunoglobulins and serum proteins in healthy humans. AB - OBJECTIVES: Link-Amster reported an increase in serum IgA when healthy subjects ingested a fermented dairy product containing Lactobacillus johnsonii La1. We aimed to assess the effects of La1 on the jejunal secretions and serum concentrations of total and specific immunoglobulins and proteins. METHODS: Twelve healthy volunteers ingested a fermented milk containing La1 or a control from day 1 till day 28, following a randomised double blind protocol. At days 0 and 28, the jejunum was successively perfused with a control solution and with a La1 suspension. The serum concentrations and jejunal secretions of albumin, orosomucoid, transferrin, alpha 2-macroglobulin, m-IgA, p-IgA, IgG, IgM, secretory component, and specific antibodies against La1 were assessed. RESULTS: Serum concentrations of IgA slightly increased between d0 and d28 in the group receiving La1 (1.85 +/- 0.64 g/L vs 1.76 +/- 0.76; P = 0.02). The other parameters were not altered. CONCLUSION: This study shows that the immunomodulating effects of La1 ingestion in man are not due to modification of jejunal protein permeability. PMID- 9207998 TI - [Pouchitis after ileal pouch-anal anastomosis]. PMID- 9207999 TI - [Hematologic manifestations related to hepatitis A virus. 3 cases]. AB - We report 3 cases of acute hepatitis A infection with haematological manifestations. In the first case, severe aplastic anemia occurred in a 6 year old child, who underwent 3 bone marrow grafts before responding favourably. In the second and third cases, severe anemia and thrombocytopenia occurred in a 42 year-old man with cholestatic hepatitis, and in a 66 year-old man with fulminant hepatitis; there was a favourable outcome in both cases. These cases demonstrate that haematological manifestations in hepatitis A can be severe, independent of the severity of liver disease. Although these manifestations seem to be rare, we suggest performing systematic haematological evaluations in cases of viral hepatitis A with unusual outcomes. PMID- 9208001 TI - [Treatment of chronic hepatitis C: doubts about a cost-effectiveness analysis]. PMID- 9208000 TI - [Acute reversible pan-dysautonomia: effect of erythromycin on gastrointestinal involvement]. AB - We describe a case of acute autonomic neuropathy in an 18-year-old woman. Gut dysfunction was sufficiently severe for the patient to undergo laparotomy for suspected mechanical-intestinal obstruction before the diagnosis was made. Apart from the gut, other organs affected included the pupils, sweat and lachrymal glands. Cardiovascular autonomic function tests showed the involvement of sympathetic adrenergic nerves. Small bowel barium X-ray showed resolution of gastric stasis and emergence of jejunum dilatation during intravenous administration of erythromycin but this treatment did not eliminate intestinal obstructive symptoms. The patient had an incomplete recovery in 3 months. Erythromycin might have therapeutic value in patients with intestinal motility dysfunction in acute dysautonomia. PMID- 9208002 TI - [Respective value of serum activity of alkaline phosphatase, gamma-glutamyl transpeptidase and 5'-nucleotidase in the diagnosis of intra- and extrahepatic cholestasis]. PMID- 9208003 TI - [Pseudocysts of the left liver: uncommon complication of acute pancreatitis]. PMID- 9208004 TI - [Acute hepatitis caused by tisopurine. A case]. PMID- 9208005 TI - [Secreting insulinoma and Bourneville's tuberous sclerosis]. PMID- 9208006 TI - [Immediate allergy to anesthetics of the amide group]. PMID- 9208007 TI - [Factitious rectorrhagia: a case of Munchausen syndrome]. PMID- 9208008 TI - [Perforation of cytomegalovirus appendicitis in a patient with human immunodeficiency virus infection]. PMID- 9208009 TI - [Guided puncture-biopsy under endosonography]. PMID- 9208010 TI - [Localization of a tumor suppressor gene distal to D22S270 in colorectal cancers]. AB - Recurrent allelic losses on chromosome 22q have been reported in colorectal cancer, distal to the NF2 gene, suggesting that another tumor suppressor gene might be involved. We report here the typing of 256 sporadic colorectal tumors and 18 colonic cancer cell lines using a set of chromosome 22 polymorphisms, ranging from 20 to 45. A panel of somatic cell hybrids, that allows to distinguish 11 bins in the 22q13 region, was used to localize 19 of the 45 selected markers and the putative tumor suppressor gene BZRP. Allelic-loss was observed in 43% of tumors. The minimal region of deletion that could be determined, telomeric to locus D22S270, refines significantly the position of the gene. The localization of the BZRP gene in this region led to a systematic screening for somatic point mutation. Direct sequencing of its coding sequence in 36 tumors hemizygous for chromosome 22 allowed the identification of three polymorphisms but failed to detect somatic mutation. PMID- 9208011 TI - [Directed mutagenesis of human recombinant receptors of vasoactive intestinal peptide of VIP1 and VIP2 type. Difference in structure-activity relationship]. AB - OBJECTIVES: Two vasoactive intestinal peptide (VIP) receptor subtypes have been cloned. We studied the structure-function relationship of human VIP1 and VIP2 receptors by mutating residues specifically conserved in extracellular domains of these receptors: N-terminal domain (E36, I43, S64, D132 and F138 in VIP1 receptor corresponding to E24, I31, S53, D116 and F122 in VIP2 receptor) and second loop (T288 and S292 in VIP1 receptor corresponding to T274 and S278 in VIP2 receptor). METHODS: Residues were mutated into alanine (A) and the corresponding cDNAs were transfected into Cos cells. Wild-type and mutated receptors were characterized in transfected cells by ligand binding assay using 125I-VIP and cAMP measurements upon VIP challenge. RESULTS: Regarding the VIP1 receptor, no specific binding of 125I-VIP could be detected on Cos cells transfected with the E36A mutant whereas other mutants, with the exception of S64A, exhibited dissociation constants similar to that of the wild-type receptor. The S64A mutant showed a 3-fold increase of its dissociation constant as compared to the wild-type receptor. cAMP experiments showed that the E36A mutant mediated a very weak stimulation by VIP. Regarding the VIP2 receptor, no specific binding of 125I-VIP could be detected on Cos cells transfected with the E24A. I31A and T274A mutants whereas all other mutants exhibited dissociation constants similar to that of the wild-type receptor. cAMP experiments showed that the E24A mutant mediated a very weak stimulation by VIP. Regarding I31A and T274A mutants, the EC50 values were increased 10 and 50 times as compared to the wild-type receptor, respectively. CONCLUSION: a) The conserved glutamate (E) residue in the N-terminal domain of VIP1 and VIP2 receptors is crucial for VIP binding; b) The VIP2 receptor contains two conserved residues isoleucine 31 and threonine 274 which are critical for VIP binding while they can be mutated without loss of function in the VIP1 receptor. This difference in the structure-function relationship should be instrumental for the development of a selective pharmacology of VIP receptor subtypes. PMID- 9208012 TI - [Gastroprotective effect of an antacid coating agent in rats. Role of endogenous prostaglandins, capsaicin-sensitive afferents neurons and nitric oxide]. AB - The aims of this study were: a) to demonstrate the gastroprotective effect of the antacid and mucosal coating agent Gastralgine (aluminium hydroxide and glycinate, magnesium trisilicate and simeticone) against ethanol- and indomethacin-induced gastric injury in the rat; b) to investigate whether gastroprotection elicited by this agent involves stimulation of capsaicin sensitive afferent neurons and activation of the nitric oxide system. METHODS: Rats received intragastrically 2 mL of the antacid or distilled water followed 1 hr later by 2 mL of 100% ethanol ig or 30 mg of indomethacin sc. The surface of ethanol-induced lesions and the length of indomethacin-induced lesions were measured. The role of afferent neurons and endogenous nitric oxide in the prevention of ethanol-induced gastric damage was determined using respectively capsaicin and the inhibitor of nitric oxide biosynthesis, NG-nitro-L-arginine. RESULTS: The antacid and mucosal coating agent significantly reduced the area of macroscopic lesions induced by ethanol and the length of lesions induced by indomethacin. Both functional ablation of afferent nerves and inhibition of nitric oxide synthesis significantly increased ethanol-induced gastric injury but failed to reverse the gastroprotective effect of the antacid against 100% ethanol. CONCLUSIONS: The antacid and mucosal coating agent Gastralgine has a gastroprotective effect against ethanol- and indomethacin induced injury in the rat. This property does not involve stimulation of capsaicin sensitive afferent neurons or synthesis of endogenous nitric oxide. PMID- 9208013 TI - [Side effects of 5-aminosalicylic acid]. PMID- 9208014 TI - [Fulminating and subfulminating hepatic failure, emergency of prevention]. PMID- 9208015 TI - [Primary prevention of digestive hemorrhage in portal hypertension]. PMID- 9208016 TI - [Imaging of atypical cysts of the liver. Study of 26 surgically treated cases]. AB - OBJECTIVES: Differential diagnosis between a benign cystic hepatic lesion, biliary cyst, and a potentially malignant lesion or biliary cystadenoma, is difficult. The aim of this study was to evaluate imaging features of atypical cystic liver lesions and the role of imaging techniques in determining a specific diagnosis. METHODS: Twenty-six patients with atypical cystic hepatic lesions were included in this study. All patients underwent surgery and pathological diagnosis was atypical hepatic cyst (n = 18), biliary cystadenoma (n = 4), hydatic cyst (n = 3), and ciliated hepatic foregut cyst (n = 1). We systematically reviewed US (n = 24), CT (n = 24), and MRI (n = 8) examinations. RESULTS: Septum were seen in both cystadenomas (US: n = 4, CT: n = 1) and hepatic cysts (US: n = 11, CT: n = 6). No mural nodules were seen. A thin wall was noted in both hepatic cysts (n = 2) and cystadenomas (n = 3). The intrahepatic biliary tract was dilated in 3 patients with hepatic cysts, 1 patient with cystadenoma, and 2 patients with hydatic cysts. Calcifications were noted in 1 patient with hepatic cyst, 3 patients with hydatic cysts, and in the case of ciliated hepatic foregut cyst. We found an associated typical hepatic cyst in 77% of cases (14/18) with atypical hepatic cysts; this was never found in other atypical cystic lesions (P < 0.01). CONCLUSION: In this series, no imaging features provided a differential diagnosis of atypical hepatic cysts and cystadenomas. The presence of associated typical hepatic cysts is helpful in suggesting the diagnosis of hepatic cyst. PMID- 9208017 TI - [Cyclosporin in the treatment of chronic diseases of the liver (excluding transplantation)]. PMID- 9208018 TI - [Fulminating and subfulminating hepatitis. Surgical aspects in management and therapeutic prospects]. PMID- 9208019 TI - [Ulcer of the esophagus disclosing probable malignant histiocytofibroma of the posterior mediastinum]. AB - We report the case of a 72-year-old woman hospitalized for dysphagia and odynophagia due to an ulcer of the esophagus. Thoracic CT-scan and esophageal endosonography revealed a tumour of the mediastinum which invaded the esophagus. Per-operative biopsies concluded to a probable malignant fibrous histiocytoma. PMID- 9208020 TI - [Hemobilia disclosing very small hepatocellular carcinoma ruptured into the biliary ducts]. AB - In hepatocellular carcinoma, invasion of the biliary tree usually occurs in large tumors. We report a case of a minute hepatocellular carcinoma which invaded the biliary tree and was revealed by secondary hemobilia, in a 65-old alcoholic patient with cirrhosis. The tumour was not identified by preoperative morphological examinations. Surgical exploration was performed due to unexplained hemobilia. Tumoral debris were observed in the right intrahepatic biliary tract during operative choledoscopy, justifying a right hepatectomy. A 5 mm diameter tumour was found in liver specimen. No recurrence had occurred after 22 months. In conclusion, in cirrhotic patients, unexplained hemobilia can reveal a small, potentially curable, hepatocellular carcinoma which has spread to the biliary tree. PMID- 9208021 TI - [Remarks on the update titled: adjuvant treatment of rectal cancer]. PMID- 9208022 TI - [Lymphocytic gastritis associated with gastric MALT lymphoma]. PMID- 9208023 TI - [Percutaneous gastrostomy: method combining digestive endoscopy and mini laparotomy]. PMID- 9208024 TI - [Late pseudo-appendicular syndrome after Roundup poisoning]. PMID- 9208025 TI - [Granulomatous involvement of the pancreas in Crohn disease associated with Takayasu arteritis]. PMID- 9208026 TI - [Recurrent acute Giardia intestinalis pancreatitis]. PMID- 9208027 TI - [Prevalence of serum anti-hepatitis E virus antibodies in patients with human immunodeficiency virus infection]. PMID- 9208028 TI - [Percutaneous alcoholization of hepatocellular carcinoma complicated by acute endocarditis on aortic valvulopathy and cerebral abscess]. PMID- 9208029 TI - [Endoscopic treatment of common bile duct calculi in patients with liver cirrhosis. Value of hydrostatic dilatation of the papilla]. PMID- 9208030 TI - Immunological activities of a lymphocyte mitogen isolated from coenurus fluid of Taenia multiceps (Cestoda). AB - The purification of a mitogen from Taenia multiceps coenurus fluid has been previously reported. In the present study, this activity, which was independent of endotoxin, stimulated the expression of lymphocyte IL-2 and Fc receptors, enhanced mitotic response to phylohaemogglutinin and concanavalin A and antagonised the previously described suppressive effects of the macrophage modifying fraction of coenurus fluid. The mitogen also increased peritoneal macrophage count and viability, Fc receptor expression and Fc receptor-mediated phagocytosis. The mitogenic activity could be destroyed by a combination of protease and amylase, but not by either enzyme alone. It is suggested that the mitogen forms part of a homeostatic mechanism for the preservation of a balanced host-parasite relationship. PMID- 9208031 TI - [Immunofluorescence reaction of antibodies directed against the intestinal epithelium of Schistosoma mansoni. VI. Immunolocalization of a carbohydrate epitope at different developmental stages]. AB - In the course of previous works, we described an IgM monoclonal antibody directed to a carbohydrate epitope located on the gut epithelium surface of the Schistosoma mansoni adult worm. We provided evidence that this epitope was present in all stages of the parasite and was particularly abundant in eggs. The current work was performed in order to specify the epitope localisation, at each stage, by immunohistochemical techniques. The epitope appears to be located on the peripheral membranes of the adult worm, while it is produced by the alive miracidium in the eggs located in the tissues and subsequently spread out inside the periovular granuloma. Moreover, in adult worms, the observed structure presents itself as a soluble form in organic solvents; on the other hand, in eggs, the epitope was essentially found made of an hydrosoluble substance. These datas can explain why, in experimentally infected mice, the epitope is mainly determined in urines at the sixth week of infestation, when eggs are settled down in the tissues. Besides, the inhibition of the monoclonal antibody fixation by a pentose which contains the Lewis X antigen, painted out that the carbohydrate structure recognised by the monoclonal antibody could be the Lewis X antigen or a very closed structure. PMID- 9208032 TI - The karyotype of Trichobilharzia franki Muller et Kimmig, 1994 (Digenea:Schistosomatidae), a new European schistosome agent of swimmers' itch. AB - Chromosome characteristics of Trichobilharzia franki Muller et Kimming, 1994, causing swimmer's itch in the Western Germany, were described and compared with the karyotype of the sympatric species Trichobilharzia szidali Neuhaus, 1952. Karyotypes of both species are very similar: diploid sets consist of seven pairs of autosome chromosomes and one pair of sex chromosomes (2n = 16, n = 5m + 2sm + Zsm/Wst), the sex determining mechanism is ZZ in males and ZW in females and gross morphology of autosome pairs does not differ markedly. The only clear discriminative feature lies in the size and shape of sex chromosomes. PMID- 9208033 TI - Comparison of three sampling methods of man-biting anophelines in order to estimate the malaria transmission in a village of south Cameroon. AB - In order to get an accurate measure of malaria transmission by mean of human bait attraction, three methods of catches for human-bail mosquitoes were compared in a Cameroonian village of high anopheline density. These three methods were 1) the classical human landing catches where the man was in the same time bait and catcher, 2) the single-nets which acted as a trap and 3) the double-nets where the outer net acted as a trap and the inner avoided mosquito bites. The anopheline densities per man with human landings were 1.6 time higher than those obtained with single-nets, these later being 4.7 times higher than those obtained with double-nets. In the three methods, the results were similar for the anophelines species catched and for their respective proportions. The samples of Anopheles nili and An. gambiae had comparative parity rates and sporozoitic indexes. From these results, in order to estimate the malaria transmission it can be envisaged to change the standard human-bait catch for the human-baited single bed-net catch; on the contrary the double-net have to be discarded because of their poor results. PMID- 9208034 TI - Role of macrophages as possible transporters of Plasmodium yoelii nigeriensis merozoites through the lymphatic system. Preliminary note. AB - Vesicles containing apparently healthy merozoites from mature schizonts were observed in the spleen and lymph nodes of mice parasitized by Plasmodium yoelii nigeriensis. They differed from all parasitic stages undergoing digestion by the macrophage and from mature schizonts of the blood. Up to 40 merozoites from three mature schizonts may be seen in the same compartment. They are thought to be accumulations of latent merozoites. PMID- 9208035 TI - Preliminary evaluation of primaquine activity on rodent malaria model after transdermal administration. AB - The aim of this preliminary study was to investigate the potential use of the transdermal route for primaquine administration in the treatment of malaria. Thus the activity of this drug on asexual blood forms of two rodent malaria parasites (P. v. petteri and P. y. nigeriensis) was evaluated following a single TTS patch application. Sustained plasma concentration values were observed for about 60 hours. The results obtained from a prepatency test showed that primaquine was more active towards P. v. petteri than P. y. nigeriensis. This preliminary study showed that the transdermal route for primaquine administration may be a promising strategy for improving the treatment of malaria in both causal prophylactic and prevention of relapses infection. PMID- 9208036 TI - Susceptibility of the cat flea, Ctenocephalides felis (Siphonaptera: Pulicidae) to four pyrethroids. AB - The amounts of active ingredient required to kill adult Ctenocephalides felis fleas on filter insecticide impregnated papers were determined in an attempt to compare the activity of different active ingredients. The following compounds were tested: bioallethrin, deltamethrin, esbiothrin and permethrin. The LD50 and LD90 against Ctenocephalides felis were 121 and 770 mg/m2 respectively for bioallethrin, and 161 and 671 mg/m2 respectively for esbiothrin. For deltamethrin and permethrin, the LD50 were 0.38 and 23 mg/m2 respectively, and 15 and 60 mg/m2 respectively for LD90. PMID- 9208037 TI - Global cerebral blood flow during infusion of adenosine in humans: assessment by magnetic resonance imaging and positron emission tomography. AB - Adenosine, an endogenous vasodilator, induces a cerebral vasodilation at hypotensive infusion rates in anaesthetized humans. At lower doses (< 100 micrograms kg-1 min-1), adenosine has shown to have an analgesic effect. This study was undertaken to investigate whether a low dose, causing tolerable symptoms of peripheral vasodilation affects the global cerebral blood flow (CBF). In nine healthy volunteers CBF measurements were made using axial magnetic resonance (MR) phase images of the internal carotid and vertebral arteries at the level of C2-3. Quantitative assessment of CBF was also obtained with positron emission tomography (PET) technique, using intravenous bolus [15O]butanol as tracer in four of the subject at another occasion. During normoventilation (5.4 +/- 0.2 kPa, mean +/- s.e.m.), the cerebral blood flow measured by magnetic resonance imaging technique, as the sum of the flows in both carotid and vertebral arteries, was 863 +/- 66 mL min-1, equivalent to about 64 +/- 5 mL 100 g-1 min-1. The cerebral blood flow measured by positron emission tomography technique, was 59 +/- 4 mL 100 g-1 min-1. All subjects had a normal CO2 reactivity. When adenosine was infused (84 +/- 7 micrograms kg-1 min-1.) the cerebral blood flow, measured by magnetic resonance imaging was 60 +/- 5 mL 100 g 1 min-1. The end tidal CO2 level was slightly lower (0.2 +/- 0.1 kPa) during adenosine infusion than during normoventilation. In the subgroup there was no difference in cerebral blood flow as measured by magnetic resonance imaging or positron emission tomography. In conclusion, adenosine infusion at tolerable doses in healthy volunteers does not affect global cerebral blood flow in unanaesthetized humans. PMID- 9208038 TI - Laser-Doppler measurements of concentration and velocity of moving blood cells in rat cerebral circulation. AB - In brain cortex all capillaries are perfused with plasma at anyone time while the flow of blood cells is heterogeneous. Increased blood flow is associated with increased number of moving erythrocytes in the microcirculation, while capillary recruitment in its classical anatomical sense appears not to exist in the brain. Modulation of the concentration of flowing erythrocytes may influence the oxygen supply to the tissue. Therefore, we examined the possibility that laser-Doppler flowmetry (LDF) could be used to quantify changes in the microvascular concentration of moving blood cells (CMBC) and blood cell velocity (< v >) by comparing LDF measurements with electromagnetic flow measurements in vitro, and confocal laser-scanning microscopy in vivo in the brain of anaesthetized male Wistar rats. In vitro measurements showed that CMBC was affected by changes in haematocrit, while < v > correlated almost linearly with blood cell velocity measured electromagnetically within a relevant physiological range. In vivo studies during hypercapnia (PaCO2 from 39 +/- 4 to 66 +/- 5 mmHg) with confocal laser scanning microscopy disclosed a 39 +/- 10% increase of cortical capillary erythrocytes, while CMBC measured with LDF increased by 37 +/- 5%. Erythrocyte flow velocity in brain cortex capillaries increased by 65 +/- 17% with confocal microscopy as compared to 72 +/- 8% with LDF. Local electrical stimulation of cerebellar cortex, and application of adenosine or sodium-nitroprusside, increased CMBC and < v > simultaneously, while during hypercapnia the < v > increase preceded the CMBC increase by 30 s. The CMBC rise rapidly reached a steady state in response to all types of stimulation, while < v > continued to increase during the major part, or the entire stimulation period. In conclusion, our data support the hypothesis that LDF may be useful for haemodynamic studies of brain microcirculation. PMID- 9208039 TI - The effects of triiodothyronine (T3) on heart rate, temperature and ECG measured with telemetry in freely moving mice. AB - We have set up a telemetry system and recorded heart rate, electrocardiogram (ECG), body temperature and the locomotor activity in awake freely moving mice. The telemetry system (DATA Sciences, St Paul, MN, USA) comprises a transmitter implanted in the peritoneal cavity and a receiver (RA1010) placed underneath the home cage. The signal from the transmitter includes the electrical activity of the heart and the body temperature. The results show that four days after the surgical procedure the mice have recovered and regained a clear circadian rhythm. The heart rate varied under baseline conditions between 432 and 618 beats min-1 and the body temperature between 35.1 and 37.7 degrees C (based on 60 min mean values). A clear time correlation between heart rate, body temperature and locomotor activity was found. As an evaluation of the method we injected T3 s.c. during a period of 4 days. Further, we were interested in whether it was possible to measure an integrated physiological response to T3 and further investigate the time course for the effect. After one day of treatment with triiodothyronine there was a significant increase in body temperature and locomotor activity. The increase in heart rate was seen after 2 days. The ECG recording revealed a significantly shortened QTend- and QRS-time. No significant difference in the PQ time was found. This method may be of great importance in studies of genetically manipulated mice. PMID- 9208041 TI - Influence of ingested fluid volume on physiological responses during prolonged exercise. AB - The effect of different rates of fluid ingestion on heart rate, rectal temperature, plasma electrolytes, hormones and performance was examined during prolonged strenuous exercise conducted at 21 degrees C. Seven well-trained males (24 +/- 1 yr; 68.6 +/- 2.9 kg; VO2 peak = 4.69 +/- 0.17 L min-1; mean +/- SEM) cycled for 2 h at 69 +/- 1% VO2 peak while receiving either no fluid replacement (NF), a volume of water estimated to prevent body weight loss (FR-100 = 2.32 +/- 0.10 L 2 h-1) or 50% of this volume (FR-60 = 1.16 +/- 0.05 L 2 h-1). The 2-h exercise bout was followed by a ride to exhaustion at a workload estimated to be 90% VO2 peak. After 2 h of exercise, NF was associated with a 3.2 +/- 0.1% weight loss, while FR-50 and FR-100 resulted in losses of 1.8 +/- 0.1 and 0.1 +/- 0.1%, respectively. Compared with FR-100, heart rate and rectal temperature were elevated (P < 0.05) during the second hour of exercise in NF, with FR-50 intermediate. Reductions in plasma volume during exercise were greater in NF and FR-50, compared with FR-100 and plasma sodium concentration was elevated in NF, decreased slightly in FR-100, with FR-50 intermediate. Plasma renin activity, aldosterone and atrial natriuretic peptide increased to similar extents in the three trials. Plasma vasopressin remained unchanged for FR-100, increased for NF, with intermediate values for FR-50. Exercise time to exhaustion at 90% VO2-peak was longer in FR-100 (328 +/- 93 s) than NF (171 +/- 75 s) with FR-50 (248 +/- 107 s) not significantly different from either FR-100 or NF. In conclusion, the responses of heart rate, rectal temperature, plasma sodium, and vasopressin during, and performance following, prolonged cycling exercise conducted at 21 degrees C are related to the amount of fluid ingested (i.e. the degree of dehydration). PMID- 9208040 TI - Rigorous swim training impairs mitochondrial function in post-ischaemic rat heart. AB - The effect of rigorous swim training (6 h day-1, 5 days week-1 for an average of 191 h) on mitochondrial respiratory function was investigated in rat heart subjected to in vivo ischaemia reperfusion (I-R). Mitochondria was isolated from the risk region of the left ventricle subjected to 60 min occlusion of the main left coronary artery followed by 30 min reperfusion. Heart weight and heart-to body weight ratio was increased by 21 and 28% (P < 0.01), respectively, in the trained (T, n = 15) vs. control rats (C, n = 20). I-R per se showed minimal effect on heart mitochondria regardless of training status. In sham, state 4 respiration rate was 26 and 32% (P < 0.05) lower in T vs. C rats, using malate pyruvate (M-P) and 2-oxoglutarate (OG) as substrates, respectively. Training also reduced state 3 respiration by 28% (M-P) and 50% (OG) (P < 0.01). The respiratory control index (RCI) was unaltered in T with M-P, but decreased with OG (P < 0.01). In vitro exposure to superoxide radicals severely reduced state 4 and 3 respiration and RCI, but T hearts showed greater reductions of state 4 and 3 rates than C. Mitochondria from T hearts also revealed a greater state 4 inhibition by H2O2 and HO. compared with C. A lower glutathione content and a higher gamma-glutamyl transpeptidase activity (P < 0.05) was observed in T vs. C. It is concluded that rigorous swim training impairs heart mitochondrial function, making them more susceptible to in vivo and in vitro oxidative stress, and that this damaging effect may be related to a diminished glutathione reserve. PMID- 9208042 TI - The effects of enalapril on the natriuretic response evoked by an oral sodium load in sodium deprived normotensive and hypertensive rats. AB - Previous studies have shown that an oral sodium load during sodium deprivation is excreted faster than an intravenous load. We wanted to study whether the renin angiotensin-aldosterone system might be associated with this phenomenon and therefore the influence of the angiotensin converting enzyme (ACE) inhibitor enalapril was investigated. The experiments were performed on four strains of rat: spontaneously hypertensive rats (SHR), Wistar-Kyoto (WKY) rats, inbred hypertension-prone (SS/Jr) and hypertension-resistant (SR/Jr) Dahl rats. In SHR and WKY rats pretreated with enalapril it was observed that an intravenous sodium load induced a renal sodium excretion which was between two and five times larger than that seen after an oral load. In SR/Jr and SS/Jr rats the sodium excretion was the same regardless of the route of administration. In SS/Jr rats sodium excretion increased three- to fourfold upon sodium repletion, whereas no significant increase was observed in SR/Jr rats. Thus, the present results indicate that an intact renin-angiotensin system is necessary for the interplay between the gastrointestinal tract and kidney. PMID- 9208043 TI - Angiotensin II induces a tachyphylactic calcium response in the rabbit afferent arteriole. AB - The influence of repeated administration of angiotensin II (AII) on smooth muscle calcium concentration ([Ca2+]i) was studied in isolated rabbit renal afferent arterioles loaded with the calcium-sensitive fluorescent probe Fura-2. [Ca2+]i was evaluated in the proximal and distal parts of the afferent arterioles. AII (10(-8) M) increased the [Ca2+]i in both these regions. A second administration of AII, however, did not elicit any response in [Ca2+]i. The response to noradrenaline administration at the end of the experiment was not affected, i.e. there was no fading or cross-desensitization. Since this desensitization was specific for AII, it was of the tachyphylaxis type. Increasing doses of AII (10( 11)-10(-8) M) did not reverse the tachyphylaxis. However, in the proximal part, pretreatment with the voltage-sensitive calcium channel blocker nifedipine (10( 6) M) blunted the tachyphylactic effect of a second administration of AII. When L arginine (L-Arg) was administered to the bath solution, thus activating the NO system, the development of tachyphylaxis was suppressed in the proximal region. Pretreatment with the protein kinase C (PKC) inhibitor chelerythrine (10(-6) M) did not affect the tachyphylaxis. We conclude that the calcium response to AII in the isolated rabbit afferent arteriole shows tachyphylaxis. This tachyphylaxis cannot be reversed by applying increasing doses of AII (10(-11)-10(-8) M). PKC does not seem to be involved in the tachyphylactic phenomenon in this preparation. It was also found that nifedipine and NO reduced the tachyphylaxis. PMID- 9208044 TI - Motor unit potential contribution to surface electromyography. AB - The background of the bioelectric activity of muscle recorded from the surface of the skin (surface electromyography) in terms of the representation of single motor units of the underlying muscle(s) is not very well documented or understood. An insight into the composition of an electromyogram is essential for the proper interpretation of one of the most widely applied electrophysiological techniques. In the present paper, a study of the contribution of single motor unit potentials to the surface electromyogram is presented. To this end, the decline of different components of the motor unit potential with depth of the motor unit is quantified. Experimentally, the action potentials from motor units at several positions in the muscle were recorded by 30 skin surface electrodes. Simultaneous use of scanning electromyography provided information about the actual position and size of the motor unit. Observed linear log-log relationships between motor unit potential magnitudes and distance indicated the usefulness of a power function to describe the motor unit potential's dependence on recording distance. It is shown that different specific surface motor unit potential characteristics fall off differently with depth. The magnitude-distance relationship is shown to be dependent on the recording configuration (unipolar vs. bipolar recording, including the inter-electrode distance) and the chosen motor unit potential parameter (negative peak amplitude, positive peak amplitude and area). PMID- 9208045 TI - Na+/glucose cotransport in the colonic adenocarcinoma cell line HT29 cl.19A: effect of cAMP. AB - The aim of the present study was to investigate the earlier finding that cAMP stimulation activates Na+/glucose cotransport in intestinal epithelia. Western blotting demonstrated the existence of Na+/glucose cotransporters in the colonic adenocarcinoma cell line HT29 cl.19A. Monolayers of this cell type showed a glucose transport, which was inhibited by 0.5 mM phlorizin (specific inhibitor of Na+/glucose cotransport). Brush border membrane vesicles of HT29 cl.19A cells exhibited a Na(+)-gradient dependent glucose transport, which was stimulated by DbcAMP-pretreatment (dibutyryladenosine 3',5'-cyclic monophosphate) of the cells. In the Ussing chamber, glucose (10 mM) unexpectedly lacked stimulatory effect on short circuit current (Isc) in HT29 cl.19A monolayers in the control situation. In DbcAMP-stimulated monolayers, glucose induced a complex Isc-response consisting of both stimulatory and inhibitory components, usually leading to a 'net' stimulation of lsc. Phlorizin (0.5 mM) did not prevent the stimulatory effect of glucose. Mannitol, alanine, fructose, ethanol (solvent for phlorizin) and the non-metabolizable glucose analogue 3-o-methyl-alpha-glucopyranoside inhibited Isc in a similar fashion as did phlorizin. Glucose transport in human colon biopsies were studied both in [14C]glucose accumulation experiments and in a specially designed Ussing chamber. There were no indications of glucose absorption in neither of these experiments. We conclude: (1) The human colon lacks Na+/glucose transport, (2) HT29 cl.19A cells exhibit Na+/glucose cotransport, which is stimulated by cAMP, (3) but this mechanism seem to be of a different type from the Na+/glucose cotransport of the 'normal' small intestine. PMID- 9208046 TI - L-arginine reverses indomethacin-induced inhibition of inflammatory fluid secretion of the feline gall bladder. PMID- 9208047 TI - Quality parameters for the production of mite extracts. AB - Mites present in house dust are of great etiological importance in type I hypersensitivity, with those belonging to the Dermatophagoides genus (D. pteronyssinus and D. farinae), of the Pyroglyphidae family, being the most frequent and principal source of allergens. For the production of allergenic extracts destined for specific diagnostic and treatment purposes of allergic diseases, the culture of such mites is absolutely necessary. In accordance with studies carried out in our laboratories to obtain adequate extracts, one must bear in mind the culture mite phase. Three growth phases have been distinguished for both species: latency phase (F1), growth phase (F2) in which the allergenic proteins are expressed with greater intensity, and death phase of the culture (F3). In the same study, the biological standardization of the extracts demonstrated that those produced from the maximum growth phase gave both in vitro and in vivo results, at least three times more sensitive than those from the other phases. We checked the reproducibility of the production method, obtaining different batches in similar conditions with a high homogeneity regarding allergenic activity. The sensitivity and specificity of the allergenic extracts depends just as much upon the production method as the standardization method. During the biological cycle of Dermatophagoides in culture, it is only from the maximum growth phase (F2), that allergenic extracts with an excellent diagnostic value, high sensitivity and specificity, can be obtained. PMID- 9208048 TI - Importance of tropomyosin in the allergy to household arthropods. Cross reactivity with other invertebrate extracts. AB - The aim of the study was to investigate the involvement of the actin binding protein tropomyosin in the allergic sensitization of patients to household arthropods, as well as to study its panallergenic character in relation to other invertebrate extracts. Three arthropod extracts were prepared, namely fly (Musca domestica), moth (Ephestia spp.) and spider (Tegenaria spp.), and used to evaluate by cutaneous and RAST tests a population of 100 household arthropod allergic patients. Twenty-nine sera were selected for the subsequent SDS-PAGE Immunoblotting assays. IgE binding bands at 36, 34, 31, 27 and 17 kDa were detected in the fly extract by more than 50% of tested sera. In moth and spider extracts, the more relevant allergens were found at 34, 31, 24 and 110, 38, 35, 26, 19 kDa, respectively. A commercial polyclonal antiserum anti-chicken muscle tropomyosin was used for tropomyosin identification. Cross-reactivity studies performed by SDS-PAGE Immunoblotting, using a pool of household arthropod allergic patients and tropomyosin antiserum, preliminarily demonstrated the presence of such protein as a cross-reacting allergen in a large variety of extracts obtained from insects, mites, crustaceans, mollusks and parasites. PMID- 9208050 TI - Allergens from Brazil nut: immunochemical characterization. AB - The increase in the consumption of tropical nuts in the Northern Hemisphere during the last years, has evolved in a simultaneous enhancement of allergic IgE mediated (Hypersensitivity type 1) reported cases produced by this kind of food. The Brazil nut is the seed of the Bertholletia excelsa tree (Family Lecythidaceae) and, as in other seeds, proteins represent one of its major components making up 15-17% of its fresh weight and 50% of defatted flour. Of these, storage proteins are the most important ones, and the 12 S globulin legumin-like protein and the 2 S albumin have been described as the most representative. The 2 S protein, due to its high sulfur-rich amino acid content (3% cysteine and 18% methionine), is being studied, cloned and expressed in some important agronomic seeds (soybean, bean, oilseed rape) in order to enrich the nutritional quality of them. The case of a patient with serious clinical allergic symptoms (vomiting, diarrhoea and loss of consciousness) caused by oral contact with the Brazil nut, is presented. The patient gave a positive Skin Prick Test response to Brazil nut, kiwi and hazelnut extracts, and negative to regionally specific aeroallergens and other food extracts. The patient serum showed a high level of specific IgE by RAST to Brazil nut (> 17.5 PRU/ml, Class 4), and significative levels to hazelnut, and mustard. In vitro immunological studies (SDS-Immunoblotting and IEF-Immunoblotting) revealed IgE-binding proteins present in the extract. It was shown that not only the heavy (Mr 9) and light (Mr 4) subunits of the known allergenic 2 S albumin but also the alpha-subunits (Mr approximately 33.5 and 32) and at least one of the beta-subunits (Mr approximately 21) of the 12 S Brazil nut globulin, hitherto never involved in allergic problems, showed a strong IgE-binding capacity. PMID- 9208049 TI - Recombinant DNA technology in allergology: cloning and expression of plant profilins. AB - Profilin, an ubiquitous protein involved in eukaryotic cytoskeleton regulation, has been previously described as allergen in grasses, weeds and trees and in many fruits and vegetables, and it is in part responsible for cross-reactivities pollen and food allergic patients. Completed cDNA clones coding for Phleum pratense, Olea europaea, Cynodon dactylon, Parietaria judaica and Helianthus annuus pollen profilins were isolated and sequenced. The deduced amino acid sequences share high identity with other plant profilins. Recombinant profilins were produced in Escherichia coli as non-fusion proteins. Induced cells produced high amounts of recombinant profilin, and after a single purification step on poly-(L-proline)-Sepharose, up to 45 mg of pure allergen per liter culture could be obtained. Recombinant profilins have similar allergenic determinants to their natural counterparts. The tertiary structure of Phleum pratense profilin described here showed three regions important for antibody recognition. The availability of a plant profilin tertiary structure opens future ways on the study of structure/antigenity relationships of these important allergens. PMID- 9208051 TI - Production and characterization of profilin monoclonal antibodies. AB - Profilins have been identified as a pan-allergen of different plant pollens and foods. In this paper, we describe the generation of monoclonal antibodies (MAbs) by immunizing BALB/c mice with Helianthus annuus purified profilin in order to characterize this important and common allergen. A panel of forty MAbs has been obtained, and twenty of them were used to map antigenic determinants in this molecule. At least two different antigenic determinants were recognized in H. annuus profilin by immunoblotting. Using the purified MAbs produced against sunflower profilin, we have analyzed the common epitope determinants in pollens of different plants: Olea europaea, Cynodon dactylon, Mercurialis annua, Phleum pratense, Parietaria judaica and Betula verrucosa. These experiments showed different cross-reactivity patterns. PMID- 9208052 TI - Fungal allergens from important allergenic fungi imperfecti. AB - Occurrence of several fungal species in the environment seems to be related to hypersensitivity disorders in humans. Fungal allergen studies reported in the literature are reviewed regarding to Aspergillus, Alternaria, Penicillium and Cladosporium genera. In this paper, we study by means of in vivo (Skin Prick Test) and in vitro (RAST and immunoblotting) the classical question of the best source of allergenic material using a population of asthmatic patients sensitized to different mould genera. In Alternaria alternata, RAST values are considerably higher in metabolic extracts (culture filtrate) than in somatic ones (myceliar). In Alternaria alternata and Penicillium chrysogenum, low molecular weight allergens (< 30 kDa) show higher IgE-binding activity in culture filtrate extracts. In Aspergillus fumigatus we find some relevant allergens in the mycellium. Different fractions have also been used in skin tests and culture filtrate extracts showing higher potency than myceliar ones, what is in agreement with the former results. We found in Penicillium chrysogenum that the 67 kDa allergen, similar to what described by Shen et al (1991), showed inespecific binding to anti-IgE conjugate used in the development of the immunoblotting technique. Whether or not this component could be a major allergen is discussed. We discuss about the importance of the biology of fungi in the sensitization and development of mould allergy. PMID- 9208053 TI - Trends in air quality in the UK. AB - Many problems are encountered when assessing trends in air quality, including changes in methods of measuring pollutants over time, different sampling timeframes, and the number and distribution of monitoring sites. Nevertheless, reasonable projections have been established for trends in sulphur dioxide (SO2), particulate matter, nitrogen dioxide (NO2), and ozone levels in the UK, which allow some estimate of future levels. The major source of SO2 is coal-fired power stations. Since the Clean Air Act of 1956, SO2 levels have decreased substantially. Nevertheless, a small but progressive increase in SO2 emissions is predicted over the next 15 years, but this is unlikely to exceed current UK air quality standards on a frequent basis. Particulate pollution (as black smoke) has also decreased dramatically since the 1960s, when the main source was fossil fuel burning. In the 1990s, the main source of particle emissions is from heavy goods vehicles. If measures currently under consideration in the UK are implemented, annual emissions may decrease to below 60 kT over the next decade, but a progressive increase is predicted thereafter. No significant changes have occurred with respect to oxides of nitrogen (NOx) and ozone levels over the last 20 years. It is predicted that NOx emissions will decline in the future due in part to the introduction of catalytic converters, while trends in ozone levels will depend on substrate supply (NOx and hydrocarbons) and weather conditions. Despite short-term trends downwards in air pollutant levels, trends may reverse in the next millennium, and continued efforts must be made to develop new ways of reducing ambient air pollutant levels. PMID- 9208054 TI - Epidemiology of allergic diseases. AB - The escalation of allergic diseases (hay fever, asthma, atopic eczema) over recent decades has been linked to an increase in environmental pollutants. The prevalence of hay fever is associated with genetic predisposition, and some reports show an association with urban areas, socioeconomic status, and combined high allergen and automobile exhaust exposure. In asthma, there is also some evidence for geographical variations in prevalence; exercise challenge tests prove positive more often in urban areas than in rural areas. Although genetic predisposition is the strongest single risk factor for atopic eczema, air pollutants may aggravate the condition by acting as unspecific irritants and immunomodulators, leading to increased immunoglobulin E expression. In a study of 678 pre-school children, the influence of maternal smoking habits on individual measures of atopy revealed a positive association between smoking during pregnancy/lactation, and a positive history of atopic eczema. An East-West German comparative study examining different types and levels of air pollution, i.e. sulphurous (industrial; East) and oxidising (urban; West), showed that the prevalence of atopic eczema was greatest in East Germany. When various direct and indirect parameters of air pollution exposure were measured, the greatest association with atopic eczema was found with NOx exposure (indoor use of gas without a cooker hood), and close proximity to roads with heavy traffic. The increased prevalence of atopic eczema cannot be explained by changes in study methodology over time, or conventional risk factors alone; environmental risk factors may be an important contributing factor. PMID- 9208055 TI - Epidemiology of pollution-induced airway disease in Scandinavia and Eastern Europe. AB - Several potential environmental factors, particularly air pollutants, have been implicated as causal factors for the increased prevalence of allergic disease in western industrialized countries. However, even when combined, these factors can only partly explain the increase. Differences in prevalence of allergy are apparent between urban and rural areas of industrialized countries, with positive skin prick tests being more common in children living in urban regions. However, the prevalence of atopy is lower in children in central and Eastern Europe, where air pollution poses a major problem, than in Western Europe. Indeed, preliminary data from the International Study of Asthma and Allergy in Children (ISAAC) confirm that the prevalence of childhood atopy is lower in Eastern Europe than in Scandinavia. Although air pollution is undoubtedly associated with the development of allergic disease, other factors connected with western lifestyle, such as changes in diet and living conditions, may play an important role and provide a possible explanation for the higher prevalence of allergic disease in western industrialized countries. PMID- 9208056 TI - Epidemiology of pollution-induced airway disease: urban/rural differences in East and West Germany. AB - The prevalence of asthma and allergic disorders was assessed in 9-11 year-old children in Leipzig and Halle in East Germany, as well as in Munich, West Germany. Both East German cities are heavily polluted due to private burning of coal and industrial emissions, while Munich has low smoke emissions but heavy road traffic. All fourth grade pupils in Munich were compared with those in Leipzig and Halle. Non-specific airway disease (bronchitis), cough, and autumn/winter nasal symptoms were most prevalent in Leipzig and Halle. Hay fever and skin test reactivity to aeroallergens were higher in West Germany compared with East Germany. Furthermore, the prevalence of asthma was also higher in the West German study area. Increased skin prick test reactivity in the West explained the increased prevalence of asthma. Longitudinal analysis showed increased respiratory symptoms on days with high mean levels of sulphur dioxide and oxides of nitrogen, as well as on days with a high peak level of 10 mu respirable particles (PM10) in East Germany. The effects of these pollutants were additive. Exposure to heavy road traffic in Munich was related to decreased pulmonary function and non-specific airway symptoms, but not to allergic sensitization and asthma. PMID- 9208057 TI - Epidemiology of pollution-induced airway disease in Japan. AB - Air pollution has been implicated as one of the factors responsible for the increased incidence of allergic diseases seen over recent years. Epidemiological studies in Japan demonstrate that atopic subjects living in urban areas are more likely to suffer from the effects of air pollution, with increased coughing, sputum production, wheezing and throat irritation. Furthermore, animal studies show that high concentrations of pollutant gases can promote airway sensitization. The incidence of allergic rhinitis and asthma have been shown to be greater in areas where there is heavy traffic and hence high levels of automobile exhaust emissions. Intranasal administration of diesel exhaust particles in mice produces a stimulatory effect on immunoglobulin E production, and a similar finding has also been shown with suspended particulate matter in air. Air pollutants, such as ozone and nitrogen dioxide (NO2), have been shown to stimulate the production of granulocyte-macrophage colony stimulating factor, which may play a vital role in airway hyperreactivity and asthma. In comparative studies of asthma in urban and rural areas, history of airway infection and a younger age of onset were found to be significantly greater in urban areas. When the asthmatic patients were divided into two groups according to environmental NO2 levels (group I: NO2 > 30 ppb; group II: NO2 < 30 ppb), no significant difference regarding the various parameters was noted between the two groups, except for a greater severity of asthma in adults in group I, and a greater severity in children in group II. These studies imply that air pollution may be one reason for the increase in allergic diseases in Japan, but a definitive conclusion cannot be drawn, and further investigation is warranted. PMID- 9208058 TI - Mechanisms of pollution-induced airway disease: in vivo studies. AB - Several studies have investigated the effects of ozone, sulphur dioxide (SO2), and nitrogen dioxide (NO2) on lung function in normal and asthmatic subjects. Decreased lung function has been observed with ozone levels as low as 0.15 ppm this effect is concentration dependent and is exacerbated by exercise. A number of lines of evidence suggest that the effect on lung function is mediated, at least in part, by neural mechanisms. In both normals and asthmatics, ozone has been shown to induce neutrophilic inflammation, with increased levels of several inflammatory mediators, including prostaglandin E2. However, in normal subjects, none of the markers of inflammation correlate with changes in lung function. The lung function changes in asthmatics may be associated with inflammatory effects; alternatively, ozone may prime the airways for an increased response to subsequently inhaled allergen. Indeed, an influx of both polymorphonucleocytes and eosinophils has been observed in asthmatic patients after ozone exposure. It has been suggested that the effect of ozone on classic allergen-induced bronchoconstriction may be more significant than any direct effect of this pollutant in asthmatics. SO2 does not appear to affect lung function in normal subjects, but may induce bronchoconstriction in asthmatics. Nasal breathing, which is often impaired in asthmatics, reduces the pulmonary effects of SO2, since this water-soluble gas is absorbed by the nasal mucosa. NO2 may also influence lung function in asthmatics, but further research is warranted. SO2 and NO2 alone do not seem to have a priming effect in asthmatics, but a combination of these two gases has resulted in a heightened sensitivity to subsequently inhaled allergen. PMID- 9208059 TI - Mechanisms of pollution-induced airway disease: in vitro studies in the upper and lower airways. AB - Evidence from both epidemiological and laboratory-based studies suggests that increased exposure to liquid petroleum and gas-derived air pollutants [nitrogen dioxide (NO2), ozone, and respirable particulate matter] may play a role in the clinical manifestation of both allergic and non-allergic airway disease. The mechanisms and cell types involved in pollutant-mediated effects in the airways, however, are not clear. In vitro studies have suggested that human fibroblasts, B lymphocytes, alveolar macrophages, and epithelial cells/cell lines may be involved. Studies of fibroblasts and macrophages have demonstrated that exposure to ozone results in decreased cell viability and increased release of pro inflammatory mediators from macrophages. Similarly, studies of B-lymphocytes have demonstrated that exposure to diesel exhaust particles (DEP) enhances the synthesis of immunoglobulin E by these cells. The airway epithelial cells have received the greatest attention in mechanistic studies of air pollution-induced airway disease and suggest that these cells are likely to play a pivotal role in the pathogenesis of airways disease. Various studies have demonstrated that exposure of nasal or bronchial epithelial cells to NO2, ozone, and DEP results in significant synthesis and release of pro-inflammatory mediators, including eicosanoids, cytokines, and adhesion molecules. Additionally, evidence suggests that epithelial cells of atopic individuals release significantly greater amounts of cytokines such as granulocyte-macrophage colony stimulating factor (GM-CSF), interleukin-6 (IL-6), IL-8, and regulated on activation, normal T-cell expressed and secreted (RANTES), on exposure to NO2 and ozone. Studies investigating the biological relevance of epithelial cell-derived pro-inflammatory mediators have shown that these enhance eosinophil chemotaxis and eosinophil adherence to endothelial cells, suggesting that pollution-induced inflammation of the airways is likely to be influenced by modulation of epithelial synthesis and release of these mediators. PMID- 9208060 TI - The role of diesel exhaust particles and their associated polyaromatic hydrocarbons in the induction of allergic airway disease. AB - The increase in allergic airway disease has paralleled the increase in the use of fossil fuels. Studies were undertaken to examine whether extracts of polyaromatic hydrocarbons (PAH) from diesel exhaust particles (DEP) (PAH-DEP) acted as mucosal adjuvants to help initiate or enhance immunoglobulin E (IgE) production in response to common inhaled allergens. In vitro studies demonstrated that PAH-DEP enhanced IgE production by tonsillar B-cells in the presence of interleukin-4 (IL 4) and CD40 monoclonal antibody, and altered the nature of the IgE produced, i.e. a decrease in the CH4'-CHe5 variant, a marker for differentiation of IgE producing B-cells, and an increase in the M2' variant. In vivo nasal provocation studies using 0.30 mg DEP in saline also showed enhanced IgE production in the human upper respiratory mucosa, accompanied by a reduced CH4'-CHe5 mRNA splice variant. The effects of DEP were also isotype-specific, with no effect on IgG, IgA, IgM, or albumin, but it produced a small increase in the IgG4 subclass. The ability of DEP to act as an adjuvant to the ragweed allergen Amb a I was examined by nasal provocation in ragweed allergic subjects using 0.3 mg DEP, Amb a I, or both. Although allergen and DEP each enhanced ragweed-specific IgE, DEP plus allergen promoted a 16-times greater antigen-specific IgE production. Nasal challenge with DEP also influenced cytokine production. Ragweed challenge resulted in a weak response, DEP challenge caused a strong but non-specific response, while allergen plus DEP caused a significant increase in the expression of mRNA for TH0 and TH2-type cytokines (IL-4, IL-5, IL-6, IL-10, IL-13) with a pronounced inhibitory effect on IFN-gamma gene expression. These studies suggest that DEP can enhance B-cell differentiation, and by initiating and elevating IgE production, may play an important role in the increased incidence of allergic airway disease. PMID- 9208061 TI - Allergen-irritant interaction and the role of corticosteroids. AB - Studies of exposure to air pollutants, such as ozone and nitrogen dioxide (NO2) +/- sulphur dioxide (SO2), have demonstrated that these agents, either individually or in combination, increase the airway response of both asthmatics and allergic rhinitics to inhaled allergen. Other studies have demonstrated that exposure to these pollutants significantly increased the levels of eosinophil cationic protein (ECP) in the nasal secretions of both asthmatics and allergic rhinitics, suggesting that pollutants may prime eosinophils for subsequent activation by allergen. More recently, our studies have demonstrated that treatment with inhaled corticosteroids, such as fluticasone propionate, significantly attenuated pollution+ allergen-induced release of ECP in allergic rhinitics. Although the mechanisms underlying the potentiating effects of pollutants on allergen-induced changes in the airways of allergic individuals are not fully understood, in vitro studies have suggested that airway epithelial cells may play an important role, since they can synthesize a variety of cytokines and adhesion molecules which influence the activity of eosinophils and other inflammatory cells. Studies of nasal epithelial cells cultured from biopsies of atopic rhinitic and atopic non-rhinitic individuals have shown that they constitutively release significantly greater quantities of pro-inflammatory cytokines than nasal epithelial cells of non-atopic individuals, and that the release of these cytokines is greater from cells of atopic rhinitics during the pollen season. Furthermore, exposure of the cells of rhinitics to ozone led to an even greater release of these cytokines, and this effect was attenuated by treatment with fluticasone propionate and beclomethasone dipropionate. PMID- 9208062 TI - Obesity in Singapore. PMID- 9208063 TI - Two-year follow-up of a behavioural weight control programme for adolescents in Singapore: predictors of long-term weight loss. AB - We followed up 112 obese adolescents enrolled in a weight control programme at a polytechnic institute in Singapore for two years. The programme used behavioural and environmental strategies to help students make sustained changes in their diet and physical activity pattern. Of these 112 students, 71.4% lost weight compared to 60% in a comparison group of 86 obese students not in the programme. About one-third (29.5%) had achieved normal weight [body mass index (BMI) < 25] with a mean weight loss of 3.3 kg among boys and 2.3 kg among girls. A stepwise multiple linear regression to determine the best combination of variables predictive of weight loss found three variables to be significantly associated with long-term weight loss: percent of weight loss in the initial 8-week intervention period; increase in exercise or physical activity following intervention, and income. These 3 variables accounted jointly for 59.5% of the variance in weight loss in males and 48% of weight loss in females. We recommend intensive efforts during the initial intervention phase to help students lose weight and strategies that can help students incorporate exercise or increased physical activity in their daily routine. PMID- 9208064 TI - Retinoblastoma in Singapore: 1976 to 1995. AB - There were at least 56 local and 42 foreign patients seen in Singapore between 1976 and 1995 inclusively. The local incidence rate is 1 in 15789 live-births, and there appears to be no predilection for sex and race. Ninety-eight per cent presented before the age of five with the average age at diagnosis being 18 months. Bilateral cases were diagnosed earlier than unilateral cases. At least 17% are hereditary, with only 2 of these with positive family histories. Of the 41 patients studied for presenting complaints, 82.9% had leukocoria, 19.5% had strabismus and 12.2% had decreased visual acuity. The main mode of treatment was enucleation alone in unilateral disease; in bilateral cases, the main mode was enucleation of the more badly affected eye together with radiotherapy and cryotherapy of the fellow eye. Ninety per cent of all patients who underwent treatment have had enucleation done. Four patients defaulted treatment--with counselling of the parents of these patients, this number could be reduced or at least kept low. Patients with early diagnosis and treatment are more likely to survive. Eight of the 56 patients died. However, with better treatment modalities, the prognosis for both life and vision have, in recent years, improved significantly. PMID- 9208065 TI - Prevalence rates of major and minor electrocardiogram abnormalities in the Singapore general population. AB - A cross-sectional survey on the general population was carried out in Singapore, with each of the 2143 persons (Chinese, Malays and Indians) aged 18 to 69 years having a 12-lead resting electrocardiogram (ECG). The tracings were coded according to the Minnesota Code with major abnormalities being Q wave changes (1 1 and 1-2), complete atrio-ventricular (A-V) block (6-1), and left bundle branch block (7-1) and minor abnormalities being lesser Q wave changes (1-3), ST depression of at least 1 mm (4-1), and T wave inversion of at least 1 mm (5-1 and 5-2). In all age groups the prevalence rates of any major abnormality were higher in males than females. In the age-adjusted 18-69 age group the prevalence rates in males and females were 2.0% and 0.5% respectively (P = 0.001). The prevalence rates of any minor abnormality (without any major abnormality) were higher in females than males in all age groups and in the age-adjusted 18-69 age group the prevalence rates in males and females were 3.5% and 5.4% respectively (P = 0.027). The prevalence rates of any minor abnormality (without any major abnormality) in the 18-29 age group in males and females were 2.2% and 4.7% respectively, indicating that these proportions of the population have minor abnormalities as a normal physiological variant. It is estimated in the 50-69 age group that 33.8% of males and 50.0% of females with any minor abnormality (without any major abnormality) have these changes as a normal physiological variant. PMID- 9208066 TI - Dobutamine stress echocardiography in the elderly Asian patients. AB - Dobutamine stress echocardiography (DSE) is an established non-invasive technique for the evaluation of coronary artery disease (CAD). It has been shown to be both safe and accurate. However, its utility and safety in the elderly, in particular, elderly Asian patients has not been studied. Between September 1992 and December 1994, we performed a total of 75 consecutive DSE studies in patients over the age of 65. Of these, 50 (67%) were females. Forty-nine patients had hypertension, 26 had diabetes mellitus, 10 were smokers, 5 had a recent or previous myocardial infarction and another 4 had a history of heart failure. Indications for DSE were, inability to perform the standard treadmill exercise test (40 patients), an abnormal resting electrocardiogram (ECG) (14 patients), a prior false positive or inconclusive treadmill test, risk stratification post myocardial infarction (4 patients) or preoperative cardiac evaluation (23 patients). The test was terminated in the majority of patients following attainment of the target heart rate. Atropine stimulation was required in 61 (81%) patients. Chest pain was provoked in 11 patients. No death or myocardial infarction occurred. Minor non cardiac symptoms occurred in another 6 patients but this did not necessitate termination of the procedure. Three patients had transient hypotension, none of which was symptomatic. Arrhythmia occurred in 23 patients but the majority were isolated atrial or ventricular premature beats (20); 1 patient had atrial fibrillation and another developed transient junctional rhythm. Only one patient developed ventricular tachycardia but this was not haemodynamically significant and terminated easily with an intravenous dose of lignocaine. A conclusive result could be obtained in 72 (96%) patients. We concluded that DSE could be performed and interpreted in the majority of elderly Asian patients studied. Despite supplemental atropine, an aggressive dosing protocol and the inclusion of patients with a myocardial scar or history of heart failure, adverse effects were rare and often did not require any specific therapy. PMID- 9208067 TI - A retrospective study on elbow function after internal fixation of intercondylar fracture of adult humerus. AB - Between 1989 and 1993, 20 intercondylar fractures of the distal humerus were treated by open reduction, internal fixation and early postoperative mobilisation. One patient died on the third postoperative day as a result of multiple injuries, leaving 19 patients for evaluation. The mean follow-up period was 4.1 years (range 2.0 to 6.7 years). According to the Muller system, there were 7 type C1 and 12 type C2 fractures. Using the Jupiter criteria, 6 elbows were rated as excellent, 9 good and 4 fair. Complications included late ulnar neuritis in one patient and wound infection in another patient. PMID- 9208068 TI - A retrospective evaluation of operative treatment of ankle fractures. AB - Between 1990 and 1992, 89 patients with ankle fracture were treated by open reduction and internal fixation in accordance with the Arbeitsgemeinschaft fur Osteosynthesefragen (AO) principles and were available for evaluation with a mean follow-up period of 3.6 years (range 3 to 5 years). According to the AO classification, there were 21 type A, 37 type B and 31 type C fractures. The results were rated by the Olerud and Molander score. Excellent or good results were seen in 20 type A, 35 type B and 25 type C fractures. Fair or poor results in 1 type A, 2 type B and 6 type C fractures. Complications included screws in the joint in 2 cases and infection in 3 cases. PMID- 9208069 TI - Necrotising fasciitis--an old enemy or a new foe. AB - Necrotising fasciitis when described by Meleney was caused predominantly by Streptococcus pyogenes. Since then, there were several reports which confirmed streptococcus as the main organism identified in this disease entity. However, recently there were reports of necrotising fasciitis caused by organisms other than Streptococcus. We analysed 19 cases of necrotising fasciitis seen in the Department of Orthopaedic Surgery over a period of 24 months to see if this disease entity has changed significantly. The patients in our series were between the ages of 19 to 85 years with an average age of 57.2 years. Males were more often affected (16 out of 19). The majority of the patients have some form of underlying disease (16 out of 19 cases). There was a trend towards polymicrobial infection and many were resistant to standard antimicrobial therapy. The mortality rate was 21.1%. Our results are comparable to many earlier series. PMID- 9208071 TI - Effects of occupational exposure to toluene: a neuropsychological study on workers in Singapore. AB - The neuropsychological functionings of workers (n = 29) occupationally exposed to low level of toluene (mean blood toluene level 1.25 ug/ml, standard deviation [SD] 0.37 ug/ml) were assessed by a test battery based on the recommendation of US National Institute of Occupational Safety and Health. The data revealed that the exposed workers performed poorer than a control group (mean blood toluene level 0.16 ug/ml, SD 0.06 ug/ml) in short-term memory, sustained attention and concentration, visual scanning, perceptual-motor speed, and finger dexterity. Results of the study confirmed that exposure to toluene may result in different degree of impairments of brain functions. PMID- 9208070 TI - Primary monosymptomatic nocturnal enuresis in Singapore--parental perspectives in an Asian community. AB - Primary monosymptomatic nocturnal enuresis (PMNE) is often not openly discussed in Asian societies. We report the parental view of PMNE in Singapore, its impact on patients and their families and the traditional beliefs and its influence on subsequent management. A screening questionnaire was used in evaluating 30 children enrolled in a clinical trial on the use of oral Desmopressin for the treatment of PMNE. Primary monosymptomatic nocturnal enuresis was familial in 56.7% of patients. Fifty per cent of them were previously unevaluated. Earlier remedial attempts included bedtime fluid restriction and voiding (100%), incentive measures (43.3%), traditional practices (26.7%), punishment (20%), drugs (16.7%), psychotherapy (100%) and bladder training (3.3%). Perceived causes of PMNE were maturational delay (50%), deep sleep (50%), familial (43.3%), behavioural problems (43.3%) and excessive fluid intake (26.7%). Reasons for seeking treatment included restricted outdoor activities (90%), parental fatigue (86.7%), disrupted sleep for the household (46.7%) and fear of underlying pathology (26.7%). Perceived adverse effects on patients included social stigma (83.3%), disrupted sleep (33.3%) and impaired school performance (13.3%). Primary monosymptomatic nocturnal enuresis can thus be a chronic distressing problem in Asian communities. PMID- 9208073 TI - Perioperative transfusion strategies: a national survey among anaesthetists. AB - A current concern in perioperative transfusion therapy is the balance between maintaining adequate haemoglobin level and yet avoiding unnecessary homologous blood transfusion. Strategies to minimise the perioperative use of homologous blood include redefining the traditional transfusion trigger of "10/30 rule", and the use of autologous transfusion therapy. Current recommendations for transfusion triggers advocate determining the "minimum acceptable haemoglobin level" for patients according to various other physiological or surgical factors. The current status of practice amongst anaesthetists in the above mentioned transfusion strategies was assessed using a national survey. An overall response rate of 59.4% was obtained. Results showed wide variation among respondents in their criteria used for preoperative and intraoperative transfusion. Recommendations on perioperative transfusion triggers made by various authors were also summarised. Autologous transfusion therapy was also not frequently practised-the reasons for this were identified. We concluded that in order to continually improve on the anaesthetic community's quality of perioperative care, continued education on the subject must be carried out. Certain practical issues also need to be addressed to facilitate the use of perioperative autologous transfusion therapy. PMID- 9208072 TI - Panic disorder in Singapore: clinical features and comparisons with generalised anxiety disorder. AB - Fifty-eight outpatients with panic disorder (PD) were examined to determine their clinical features in comparison with a cohort of 52 patients with generalised anxiety disorder (GAD). Both groups were of comparable age, sex, educational level, marital status and ethnicity. PD patients were more likely to complain of palpitations, breathlessness, chest pain, numbness, choking sensations and especially fear of dying. GAD patients tended to complain of feeling tense, insomnia, headaches, weakness, restlessness and muscle aches. PD patients had greater comorbidity especially with agoraphobia and depression. Contrary to other reports, there were more males than females in both groups but alcohol dependence and suicide attempts were relatively rare. PD symptoms seemed more distressing, caused more social and occupational disruption, led to more requests for medical investigations and earlier psychiatric consultations. These factors seemed to suggest that panic disorder is a more severe illness than generalised anxiety disorder. PMID- 9208074 TI - Sedation for the conduct of lumbar epidural anaesthesia: a study using subanaesthetic dose of ketamine in combination with midazolam. AB - We gave sedation for the conduct of lumbar epidural analgesia using intravenous ketamine 0.3 mg.kg-1 with intravenous midazolam 2 mg. Forty adult Chinese females undergoing major gynaecological laparotomies had epidural catheter inserted before general anaesthesia, 20 of whom were given ketamine and midazolam (study group) and the other 20 acted as control. During the conduct of the epidural, the pain and anxiety scores in the study group were significantly less than the control group (P < 0.05). Patients were significantly more sedated in the study group (P < 0.05). All the patients in the study group were satisfied and would consent to future epidural versus 75% in the control group (significant at P < 0.05). Ninety per cent of patients in the study group had amnesia but none in the control group. Pain experienced during the epidural was the reason for refusal of future epidural. We did not observe any emergence phenomenon or cardiovascular stimulation. There was a statistically significant decrease in the pulse oximetry oxygen saturation (SpO2) in the study group but none required oxygen supplementation. We concluded that pain caused by the conduct of epidural did decrease the patient's acceptance rate to future epidural, and the combined use of intravenous ketamine 0.3 mg.kg-1 and midazolam 2 mg provided adequate sedation, analgesia, anxiolysis and amnesia to significantly increase the acceptance rate without any significant side effects. PMID- 9208075 TI - The safety of weekly low dose oral methotrexate in an Oriental population with rheumatoid arthritis. AB - A retrospective review of 50 oriental patients with rheumatoid arthritis treated with weekly oral methotrexate showed adverse events in 15 (30%) patients with 19 occurrences (38%) of leucopaenia (4%), pancytopaenia (2%), gastrointestinal symptoms (18%), hepatic transaminase elevation (6%), rash (2%) and infections (6%). The median duration of treatment with methotrexate was 11 months (range 1 to 105 months). Pancytopaenia occurred in 1 patient with renal failure. All adverse events resolved with cessation of therapy and on several occasions, despite continued therapy. Methotrexate was discontinued permanently in 2 and temporarily in 7 patients as a result of adverse events. No recurrence of adverse events was noted on restarting methotrexate therapy in patients with non life threatening adverse events. No increase in adverse events was noted in 14 patients treated with a combination of methotrexate and anti-malarial therapy. We conclude that methotrexate was well tolerated by the Oriental patients with rheumatoid arthritis in our study and could be safely restarted in those patients with non life-threatening adverse events. PMID- 9208076 TI - A retrospective study of 13 Oriental children with juvenile dermatomyositis. AB - This is a retrospective study of 13 Oriental children with juvenile dermatomyositis (JDM). We reviewed data found in the hospital records of children diagnosed to have definite (n = 4), probable (n = 7) and possible (n = 2) JDM who presented over a 10-year period at 4 centres in Singapore and compared our results with the experience of others. We found an overall female preponderance (female to male ratio of 3.3:1) but an equal sex ratio in children below 5 years of age. The majority (92%) had insidious onset and good outcome. Diagnosis was often delayed because of the insidious onset, and because weakness occurred late, was mild or absent. Only one child had an acute presentation and refractory course. She died despite aggressive therapy. Clinical features, complications and mainstay medication used were similar to Western studies. PMID- 9208077 TI - Screening for and treatment of hypercholesterolaemia--a review. AB - The screening for and treatment of hypercholesterolaemia is still a complex and controversial issue. As hypercholesterolaemia is a major risk factor for coronary heart disease (CHD), there was initially much enthusiasm for mass screening with intervention recommended for persons with serum cholesterol levels > or = 6.5 mmol/L. However, trials of treatment of hypercholesterolaemia using lipid lowering drugs showed overall benefit (i.e. reduced all-cause mortality) only for persons with pre-existing CHD (secondary prevention) or other major risk factors. For this and other reasons (e.g. cost effectiveness) screening for hypercholesterolaemia has been recommended for persons with CHD or major risk factors (family history of premature coronary heart disease, cigarette smoking, hypertension, and diabetes mellitus). Recent trials with statins have shown greater benefit than with other lipid lowering drugs, confirming the benefits of secondary prevention. In addition, one trial has shown overall benefit after 5 years for middle-aged hypercholesterolaemic males without a previous myocardial infarction, raising the possibility of mass screening. However, longer follow-up of a number of trials of primary prevention including on persons without risk factors together with a full assessment of the local situation with regards to hypercholesterolaemia is needed before mass screening can be recommended. PMID- 9208078 TI - Coincident cold urticaria in a married couple: case report and a literature review. AB - We describe an interesting coincidence in which a woman developed acquired idiopathic cold urticaria and her husband systemic cold urticaria eight months later. The occurrence of cold urticaria in a tropical country like Singapore is rare. Each case illustrates the typical features of that particular type of cold urticaria, including the appropriate response to the challenge test and its self limited nature. PMID- 9208079 TI - Chylothorax: case report and review of literature. AB - Chylothorax is a relatively uncommon condition. We report a case of a 53-year-old Chinese man with chylothorax who presented with cough, breathlessness, loss of weight and generalised lymphadenopathy. He was initially diagnosed as having tuberculosis based on histology and cultures of the cervical lymph node. However, immunophenotyping of the pleural fluid by flow cytometry showed the presence of a monoclonal B cell population with Kappa light chain expression suggesting an underlying lymphoproliferative disorder. A repeat lymph node biopsy showed malignant lymphoma (follicular small cleaved type). In spite of aggressive measures which included bilateral tube thoracostomy drainage, a low fat diet, medium chain triglyceride diet, total parenteral nutrition, chemotherapy and mediastinal irradiation, his chylothorax persisted. Hence, we report a patient with persistent chylothorax and generalised lymphadenopathy who was subsequently diagnosed to have concurrent tuberculosis and malignant lymphoma. The literature on chylothorax is reviewed. PMID- 9208080 TI - Pseudomyopia in a patient with blocked ventriculo-peritoneal shunt--a case report. AB - Accommodative spasm usually encompasses a classical triad of pseudomyopia, esodeviation and pupillary constriction. Accommodative spasm is most often psychogenic in nature; however, it may be associated with other organic diseases of which a rare cause is that of intracranial catheter complications. We report a case of dorsal midbrain syndrome with pseudomyopia in a patient with a blocked ventriculo-peritoneal shunt inserted for aqueductal stenosis. Clinical presentation was unusual in this patient as pseudomyopia occurred with exodeviation and without pupillary constriction. PMID- 9208081 TI - Monoplegia following obstetric epidural anaesthesia. AB - We describe a case of transitory monoplegia following obstetric epidural anaesthesia which we believe to be either due to direct trauma to the nerve roots or radicular artery spasm resulting in spinal cord ischaemia both secondary to the epidural catheter. An obstetric epidural was performed on a 34-year-old lady with no significant past medical history. Transient radicular pain down her left leg was experienced during the insertion of the catheter. She had unilateral block on her left side which could not be rectified despite injecting local anaesthetics in the right lateral position, resiting the epidural catheter, and administering a total dose of 6 ml of 1.5% plain lignocaine and 19 ml of 0.25% plain bupivacaine. There was no hypotension, and no vasoconstrictor was used epidurally. She still had left-sided numbness and weakness 9 hours after removal of the catheter, but without urinary or bowel dysfunction. A magnetic resonance imaging scan excluded any space occupying lesion. She was managed conservatively and had complete recovery 35 hours after the last dose of local anaesthetics. PMID- 9208082 TI - Malignancy arising in seborrheic keratosis: a report of two cases. AB - We describe the pathological features of two cases of malignant transformation occurring in seborrheic keratosis. The lesions affected elderly individuals: a 79 year-old Chinese lady and a 79-year-old Chinese gentleman, both of whom had long standing seborrheic keratoses with recent increase in size. Histologically, the malignancies identified were invasive well-differentiated squamous cell carcinoma and Bowen's disease (squamous carcinoma in situ), respectively. Pre-existing seborrheic keratoses demonstrating a zone of transition into the malignant foci were present. Aetiologic factors that have been proposed to explain these exceedingly rare occurrences have included prolonged sun damage and chronic low dose radiation exposure. PMID- 9208083 TI - Sclerosing lipogranuloma of the scrotum following a laparoscopic herniorraphy and varicocelectomy--a case report. AB - "Sclerosing lipogranuloma" was first coined to describe a lesion of adipose tissue thought to be due to trauma. Later, analysis of the fat within these lesions showed the presence of exogenous paraffin hydrocarbons and a link with paraffin injection was postulated. In Japan, lipogranulomas without a history of trauma or paraffin injection have been described more frequently. Our patient, developed such a lesion in his scrotum following a laparoscopic left herniorrhaphy and varicocelectomy (intra-abdominal approach). He had no prior history of trauma or paraffin injection. A possible relationship is postulated. PMID- 9208084 TI - Postgraduate surgical training--an era of subspecialisation: qua vadis (where are you going?). PMID- 9208085 TI - Surgical training in Indonesia: a plan for change. PMID- 9208086 TI - Undergraduate surgical training--where are we heading? PMID- 9208087 TI - Career opportunities for medical specialisation in the 21st century. PMID- 9208088 TI - Factors influencing perfect surgical outcome. AB - With affluence and education, the population of Asia will be demanding quality surgical care. The energetic, affluent and educated Asian professionals and business communities in the cities demand the best; and in surgery, they seek perfect results. Perfect results require a combination of 3 factors: the skill, knowledge and experience of the surgeon. He must be a skilled surgeon with good basic surgical techniques and also technical skills in the management of his discipline combined with meticulous attention to details. Furthermore, he must have a clear knowledge of the basic physiopathology of surgical principles of the condition he is to manage. Experience with difficult situations and intrasurgical problems are essential for success. PMID- 9208089 TI - Seventh Seah Cheng Siang Memorial Lecture. The aetiology of gallstones. PMID- 9208090 TI - Conformational studies of dithymidine boranomonophosphate diastereoisomers. AB - The boranophosphate ester nucleotides are a new class of nucleic acid analogues that are isoelectronic and isostructural to normal phosphodiester nucleic acids and that maintain the anionic charge of the nucleic acid backbone. The two P diastereoisomers of dithymidine boranomonophosphates were separated using reverse phase HPLC; the faster and slower eluting isomers are designated as d(TpBT)-1 and d(TpBT)-2, respectively. Conformations of the isomers were studied using-circular dichroism (CD) and NMR, and compared to the analogous phosphate diester, d(TpT). This comparison allowed the effects of the borane group and chirality of the boranophosphate linkage on sugar and base conformations to be assessed. The CD spectra of the diastereoisomers are consistent with both having a B-type conformation. Analysis of the 1H-1H and 1H-31P coupling constants showed that these conformations are similar to those of the unmodified parent dimer; specifically, the 2'-deoxyribose rings prefer the S (C2'-endo) conformation, and the C4'-C5' and C5'-O5' rotamers are primarily in the gamma + and beta + conformations, respectively. Conformational differences between the diastereoisomers and between the modified and unmodified dimers are manifested by differences in the preferences of the 3'-residues to adopt S sugar pucker and beta + conformations. There is reduced preference for the S sugar pucker of the 3'-residue in d(TpBT)-1 relative to d(TpBT)-2, which is similar to d(TpT). There is less preference for the beta + conformation of the 3'-residue in d(TpBT)-2 relative to d(TpBT)-1 and d(TpT). Based on the CD results, the temperature dependences of the thymidine H6 chemical shifts, and the derived sugar ring and backbone conformational parameters, we conclude that the borane group exerts a minimal influence on the sugar conformations and base stacking interactions. Preliminary assignment of the absolute configuration of the pair of SP and RP diastereoisomers to d(TpBT)-1 and d(TpBT)-2, respectively, is made on the basis of enzyme selectivity and NOE difference experiments. PMID- 9208091 TI - In vitro and ex vivo inhibition of hepatitis A virus 3C proteinase by a peptidyl monofluoromethyl ketone. AB - Hepatitis A virus (HAV) 3C proteinase is the enzyme responsible for the processing of the viral polyprotein. Although a cysteine proteinase, it displays an active site configuration like those of the mammalian serine proteinases (Malcolm, B. A. Protein Science 1995, 4, 1439). A peptidyl monofluoromethyl ketone (peptidyl-FMK) based on the preferred peptide substrates for HAV 3C proteinase was generated by first coupling the precursor, N,N-dimethylglutamine fluoromethylalcohol, to the tripeptide, Ac-Leu-Ala-Ala-OH, and then oxidizing the product to the corresponding peptidyl-FMK (Ac-LAAQ'-FMK). This molecule was found to be an irreversible inactivator of HAV 3C with a second-order rate constant of 3.3 x 10(2) M-1 s-1. 19F NMR spectroscopy indicates the displacement of fluoride on inactivation of the enzyme by the fluoromethyl ketone. NMR spectroscopy of the complex between the 13C-labeled inhibitor and the HAV 3C proteinase indicates that an (alkylthio)methyl ketone is formed. Studies of polyprotein processing, using various substrates generated by in vitro transcription/translation, demonstrated efficient blocking of even the most rapid proteolytic events such as cleavage of the 2A-2B and 2C-3A junctions. Subsequent ex vivo studies, to test for antiviral activity, show a 25-fold reduction in progeny virus production as the result of treatment with 5 microM inhibitor 24 h post-infection. PMID- 9208092 TI - Alpha 1-adrenoceptor subtype selectivity: molecular modelling and theoretical quantitative structure-affinity relationships. AB - This study constitutes a preliminary rationalization, at the molecular level, of antagonist selectivity towards the three cloned alpha 1-adrenergic receptor (alpha 1-AR) subtypes. Molecular dynamics simulations allowed a structural/dynamics analysis of the seven alpha-helix-bundle models of the bovine alpha 1a-, hamster alpha 1b-, and rat alpha 1d-AR subtypes. The results showed that the transmembrane domains of these subtypes have different dynamic behaviours and different topographies of the binding sites, which are mainly constituted by conserved residues. In particular, the alpha 1a-AR binding site is more flexible and topographically different with respect to the other two subtypes. The results of the theoretical structural/dynamics analysis of the isolated receptors are consistent with the binding affinities of the 16 antagonists tested towards the three cloned alpha 1-AR subtypes. Moreover, the theoretical quantitative structure-affinity relationships obtained from the antagonist-receptor interaction models further corroborates the hypothesis that selectivity towards one preferential subtype is mainly modulated by receptor and/or ligand distortion energies. In other words, subtype selectivity seems to be mainly guided by the dynamic complementarity (induced fit) between ligand and receptor. On the basis of the quantitative models presented it is possible to predict both affinities and selectivities of putative alpha 1-AR ligands as well as to estimate the theoretical alpha 1-AR subtype affinities and selectivities of existing antagonists. PMID- 9208093 TI - Detection and structural characterization of ras oncoprotein-inhibitors complexes by electrospray mass spectrometry. AB - MS based methodology employing electrospray ionization (ESI) is described for the detection of ternary complexes in which SCH 54292 or SCH 54341 and GDP are noncovalently bound to oncogenic ras protein. The observed molecular weights of 19,816 and 19,570 Da confirmed the presence of noncovalent complexes of ras-GDP SCH 54292 and ras-GDP-SCH 54341, respectively. We have also performed selective chemical modification of lysine residues of the ras protein complex followed by enzymatic digestion and on-line LC-ESI MS peptide mapping to determine protein drug binding topography. There was a good correlation between nucleotide exchange inhibition as determined by the enzyme assay and evidence of complex formation as determined by MS. PMID- 9208094 TI - Stereochemistry of yeast delta 24-sterol methyl transferase. AB - S-Adenosyl-l-methionine: delta 24-sterol methyl transferase (24-SMT) mediates introduction of the C-28 carbon of yeast sterols. It has been shown that sulfonium analogues of the presumptive cationic intermediates of the methylenation reaction are potent in vivo and in vitro inhibitors of this process. In the presence of these inhibitors, cultures of yeast produced increased proportions of zymosterol, the natural substrate of the enzyme, while proportions of ergosterol and ergostatetraenol were decreased. New C27-sterol metabolites were also found. The in vivo inhibitory power of the analogues [I50 (microM)] was determined from the proportion of C-24 methylated sterols to C-24 nonmethylated sterols in treated cultures to be in the following order: 25 thiacholesterol iodide (0.07) > 24(S)-methyl-25-thiacholesteryl iodide (0.14) > 24(R)-methyl-25-thiacholesteryl iodide (0.25). Kinetic inhibition as revealed by radiolabeled S-adenosyl-l-methionine (SAM), crude enzyme and 25-thiacholesteryl iodide revealed this inhibitor to be uncompetitive with respect to zymosterol and competitive with respect to SAM. The greater inhibitory power of 24(S)-methyl-25 thiacholesteryl iodide compared to 24(R)-methyl-25-thiacholesteryl iodide suggests that methyl donation to delta 24 occurs from the si face. When considered in conjunction with Arigoni's previous work, the present results infer the methylenation mediated by yeast 24-SMT proceeds by alkylation from the si face of delta 24 followed by migration of a hydrogen from C-24 to C-25 across the re face and final loss of a hydrogen from C-28 on the re face. PMID- 9208095 TI - Imidazoline receptors: qualitative structure-activity relationships and discovery of tracizoline and benazoline. Two ligands with high affinity and unprecedented selectivity. AB - The observation that all the attempts to characterize imidazoline (I) receptors have been carried out with non-selective or poorly selective ligands prompted us to undertaken research aimed at developing selective ligand(s). In previous work using, as a starting point, cirazoline I, a potent alpha 1-adrenergic receptor agonist that also binds to I receptors, we showed that removal of the cyclopropyl ring (2) retains high affinity for I2 receptors while reducing alpha 1-adrenergic agonist activity. However, it was felt that this residual, albeit modest, alpha 1 adrenergic agonist activity might diminish the usefulness of compound 2, and we now report on our continuing efforts in this field. Starting from compound 2, we first eliminated the alpha 1-agonist component by isosteric replacement and then, by means of conformational restrictions on compound 7, succeeded in discovering tracizoline (9) and benazoline (12). These two new ligands with high affinity (pKi value 8.74 and 9.07, respectively) and unprecedented selectivity with respect to both alpha 2- (I2/alpha 2 7,762 and 18,621) and alpha 1- (I2/alpha 1 2,344 and 2,691) adrenergic receptors, are valuable tools in the study of I receptor structure and function. In addition, the large number of derivatives studied has allowed us to establish congruent qualitative structure-activity relationships and identify some structural elements governing affinity and selectivity. PMID- 9208096 TI - 2-D and 3-D modeling of imidazoline receptor ligands: insights into pharmacophore. AB - A 3-D quantitative structure-activity relationship (3-D QSAR) study was carried out using comparative molecular field analysis (CoMFA) on both imidazoline (I2R) and alpha 2 receptor binding affinities of a large series of 2-substituted imidazolines. Significant cross-validated correlations, having promising predictive ability, were obtained along with 3-D pharmacophore models that defined the spatial regions where steric and electrostatic interactions may modulate the in vitro binding affinities and indicated possible physicochemical and structural requirements for I2/alpha 2 receptor selectivity. PMID- 9208097 TI - Further syntheses employing phosphorylase. AB - Maltopentaose was immobilized on silica gel and used as a recyclable primer in the reaction of glycogen phosphorylase with D-glucal. An improved method to obtain 2-deoxy-alpha-D-arabino-hexopyranosyl phosphate (4) by using this catalyst as well as the specific synthesis of low molecular weight, water-soluble 2-deoxy maltooligosaccharides (12, 13, 14 and 15) are described. Further investigations with modified phosphorylase substrates showed that mannosyl phosphate (16) can be slowly transferred to the primer maltotetraose. alpha-1,4-Mannosyl-maltotetraose (17) and its degradation product alpha-1,4-mannosyl-maltose (18) were identified. PMID- 9208098 TI - Antioxidative constituents in Heterotheca inuloides. AB - Sesquiterpenoids, 7-hydroxy-3,4-dihydrocadalin and 7-hydroxycadalin, and flavonoids, quercetin, kaempferol and their glycosides, isolated from Heterotheca inuloides (Asteraceae), a Mexican medicinal plant known as "arnica", were evaluated as antioxidants. These compounds showed potent scavenging activity on diphenyl-p-picrylhydrazyl (DPPH) radical. Microsomal lipid peroxidation induced by Fe(III)-ADP/NADPH was inhibited by both terpenoids and flavonoids, though only flavonoids possessed superoxide anion scavenging activity in microsome. Flavonoids also scavenged enzymatically and non-enzymatically generated superoxide anion. On the other hand, mitochondrial lipid peroxidation induced by Fe(III)-ADP/NADH was inhibited only by sesquiterpenoids. Furthermore, these terpenes protected mitochondrial enzyme activity against oxidative stress. These results showed that two types of antioxidants existed in the dried flower of H. inuloides and would contribute to protection of tissues against various oxidative stresses. PMID- 9208099 TI - Feasibility of an immunoassay for mevalonolactone. AB - Mevalonic acid is a key intermediate in a broad spectrum of cellular biological processes and their regulation. Availability of a rapid, sensitive and accurate method for its assay would be highly useful. Therefore, the feasibility of developing an immunoassay for mevalonic acid in biological samples was explored. The strategy employed was to synthesize several racemic haptens structurally resembling R-mevalonolactone, the cyclic form of mevalonic acid present at lower pH and presumed to be more antigenic. Two of these haptens were coupled to keyhole limpet hemocyanin, and the resulting conjugates were used successfully to generate antibodies in rabbits. The first antiserum bound to R,S-mevalonolactone much more effectively at pH 4.0 than at pH 6.0, consistent with the structural resemblance of the haptens to the lactone form. This antiserum also bound the free hapten from which it was generated and two others of different structure with comparable effectiveness; and slightly better than it bound R,S mevalonolactone at pH 4.0. Similar results were obtained with the antiserum to the second hapten. The binding of either antiserum to the natural enantiomer, R mevalonolactone, was 20 times weaker than to R,S-mevalonolactone, suggesting that the nonbiological enantiomer was more antigenic. Nevertheless, the results demonstrate that an immunochemical approach to accurate quantitation of mevalonic acid in biological samples is feasible. PMID- 9208100 TI - Structure-activity relationships of cyclic enediynes related to dynemicin A. I. Synthesis and antitumor activity of 9-acetoxy enediynes equipped with aryl carbamate moieties. AB - A series of the 9-acetoxy enediyne compounds, 6a-k which were simplified from natural dynemicin A, and designed to be equipped with various aryl carbamate moieties, was synthesized and evaluated for DNA-cleaving ability, in vitro cytotoxicity, and in vivo antitumor activity. As a result of this study of the structure-activity relationships (SAR) with regard to the Rt substituent, both compounds 6a and 6f with the phenyl carbamate and 4-chlorophenyl carbamate moiety, respectively, were found to exhibit significant activity (T/C > 200%) against murine P388 leukemia in mice, in spite of having IC50 values in the micromolar range. In particular, compound 6f showed the most potent activity with a maximum T/C of 256% at a daily dosage of 4.0 mg/kg for four days. Furthermore, both compounds 6a and 6f were effective against Meth A sarcoma in mice and inhibited 71 and 77% of the tumor growth at 2.0 and 3.0 mg/kg dosages, respectively. In contrast to 6f, compound 6i possessing the 2-nitrophenyl carbamate moiety showed only a slight in vivo activity, while it had about one order of magnitude higher in vitro cytotoxicity than 6f. For the stereochemistry activity relationships at the C9 position, the (9R*)-isomers of 6c, 6g, and 6j were found to show higher in vitro and in vivo potencies than the corresponding (9S*)-isomers. PMID- 9208101 TI - Structure-activity relationships of cyclic enediynes related to dynemicin A. II. Synthesis and antitumor activity of 9- and 12-substituted enediynes equipped with aryl carbamate moieties. AB - Novel enediyne compounds 4-8, simple analogues of dynemicin A (1) equipped with the phenyl or 4-chlorophenyl carbamate moiety, were synthesized and evaluated for DNA-cleaving ability, in vitro cytotoxicity, and in vivo antitumor activity. As a result of the SAR study, it was revealed that the size and character of the substituents (R1 and R2) at the C9 position critically influenced both the stability and antitumor activity of the enediyne compounds. We found that the 9 deoxy compound 6a, a stable and less bulky enediyne having a hydrogen as the R1 and R2 substituents, showed a significant in vivo activity with a T/C of 215% at a daily dosage of 2.0 mg/kg for 4 days. The incorporation of an oxygen-containing functional group as the R3 substituent on a benzene ring resulted in considerable abolishing of both the in vitro and in vivo potencies. In a series of 9-acyloxy compounds, incorporation of the basic aromatic moiety such as 8e was effective for the in vitro activity, but it was ineffective for the in vivo activity. Furthermore, for the stereochemistry-activity relationships at the C9 position, the (9R*)-isomers of 8c, 8e, and 8f were found to show higher both in vitro and in vivo than the corresponding (9S*)-isomers. For the mechanistic studies, compound 6a underwent Bergman cycloaromatization via a diradical pathway under acidic conditions, whereas it scarcely showed DNA-cleaving activity due to the chemical stability of the aryl carbamate moiety under neutral conditions. PMID- 9208102 TI - Synthesis and biological evaluation of potent glycosidase inhibitors: N-phenyl cyclic isourea derivatives of 5-amino- and 5-amino-C-(hydroxymethyl)-1,2,3,4 cyclopentanetetraols. AB - Twenty-four stereoisomers of 5-amino- and 5-amino-C-(hydroxymethyl)-1,2,3,4 cyclopentanetetraols and twenty-six of the corresponding N-phenyl cyclic isourea derivatives were assayed for inhibitory activity against six glycosidases. Among them, as has been expected for structure mimics of putative transition state glucopyranosyl cation for glycoside hydrolysis, 1L-(1,2,4,5/3)-5-amino-1-C (hydroxymethyl)-1,2,3,4-cyclopentanetetrao l L-4 and its N-phenyl cyclic isourea derivative S-19 were shown to have strong inhibitory activity, IC50 4 x 10(-7) and 7.6 x 10(-9) M, respectively, against baker's yeast alpha-glucosidase. It has been analogously explained that compounds R,S-22 and R,S-26 possessed high inhibitory potency against Escherichia coli and bovine liver beta-galactosidases, respectively. PMID- 9208103 TI - Synthesis and biological activity of A-nor-paclitaxel analogues. AB - A number of paclitaxel analogues with a 5-membered A-ring (A-nor-paclitaxels, or (15-->1)-abeo-paclitaxels) have been prepared in order to determine whether analogues of this class might have improved bioactivity as compared with paclitaxel. Most of the compounds synthesized were less active than paclitaxel, but one analogue was equivalent to paclitaxel in a tubulin-assembly assay, and another analogue was more cytotoxic than paclitaxel in two different cell lines of the NCI screen. PMID- 9208104 TI - Molecular modeling of (E)-1-alkyl-4(3)-[2-(1H-azolyl)vinyl]-pyridinium salts and evaluation of their behavior towards choline acetyltransferase. AB - A new type of extended pi-system aza-analogue of (E)-4-[2-(1-naphthylvinyl)]-1 substituted pyridinium salts (NVP+) has been designed and its inhibitory activity towards choline acetyltransferase (ChAT) has been evaluated in vitro. Among the several examples of the title quaternary salts synthesized 2 and 3, the indolylvinylpyridinium salt 2e is the only one to show a very low ChAT inhibition. The molecular modeling study is highly illustrative of their behavior towards ChAT and interaction with the recognition site. Thus, several selected cations together with the reference NVP+ compound 1a were studied at the PM3 and AM1 levels respectively. At the global minima, all the compounds are planar, which, from the electron charge distribution, shows a degree of polarization similar to the NVP+ model compound 1a. However, the fitting of all optimized structures indicated that only the indole derivative 2e showed the same aromatic fragment orientation as 1a, which allows us to define a volume that is not accessible to ligands in the enzyme and consequently to a refined model of the choline acetyltransferase recognition site. PMID- 9208105 TI - Antiviral and tumor cell antiproliferative SAR studies on tetracyclic eudistomins. II. AB - In a search for the minimum pharmacophore of the naturally occurring tetracyclic eudistomins, five structural analogues (4-8) were evaluated for their in vitro antiviral and tumor cell antiproliferative activities. For the synthesis of these derivatives both intra- and intermolecular Pictet-Spengler reactions have been used. Opening of the beta-carboline annulated 7-membered D-ring in 6 and 7 resulted in a complete loss of activity. On the other hand, replacement of either the oxygen atom or the sulfur atom in the 7-membered ring by a methylene group in 5 and 8, respectively, is allowed. These results combined with previous SAR data underline the crucial importance of the D-ring in eudistomins as a scaffold for the correct positioning of both basic nitrogen atoms. Also bioisosteric replacement of the bicyclic indole system with a dimethoxyphenyl group, to give the isoquinoline skeleton, is allowed. The tricyclic isoquinoline derivative 4 is, so far, the most promising antiviral analogue; it combines a high potency (MIC at 100 ng/ mL (340 nM)) with high MCC/MIC ratios (ranging from 1000 to 5000 against HSV-1, HSV-2, vaccinia virus, and vesicular stomatitis virus. PMID- 9208106 TI - New serine protease inhibitors with leukotriene B4 (LTB4) receptor binding affinity. AB - A series of new trypsin-like serine protease inhibitors, 1, 2 and 7-23, containing amidinobenzene moiety was found to show potent LTB4-receptor affinity. Among them, compounds 1 and 2 were found to be LTB4 receptor antagonists based on an inhibition assay of human polymorphonuclear neutrophil (PMN) intracellular calcium mobilization induced by LTB4. Compounds 1 and 2, which satisfy the reported structural requirements for good oral activity, are expected to show a balanced dual mode of action, i.e., protease inhibitory activity and LTB4 receptor antagonist activity, in vivo. PMID- 9208107 TI - Synthesis and antitumor activity of water-soluble enediyne compounds related to dynemicin A. AB - The enediyne compounds 9-14, simple dynemicin A (1) analogues equipped with aryl carbamate moieties with various aliphatic amino or hydroxy groups at the C9 position, were synthesized and evaluated for DNA-cleaving ability, in vitro cytotoxicity, and in vivo antitumor activity. We found that the water-soluble compounds, in which the tert-amines such as the 2-(dimethylamino)ethyl (10b, 14b), 2-(pyrrolidino)ethyl (10c), or 1-azabicyclo[3.3.0]oct-5-ylmethyl (10d, 12d, 14d) group were attached, showed not only the enhanced in vivo antitumor activity but also the decreased toxicity compared to the corresponding 9-acetoxy enediyne compounds 6-8. In particular, compound 10c showed the most enhanced in vivo antitumor activity (T/C = 222% at a daily dose of 1.25 mg/kg for 4 days) at about half of the dose of 6. These results suggest that both the enhanced antitumor activity and the reduced toxicity might be due to the improved bioavailability or disposition of compounds 6-8 by their water-solubilization. PMID- 9208108 TI - Transplant of bone marrow and cord blood hematopoietic stem cells in pediatric practice, revisited according to the fundamental principles of bioethics. AB - The two most widely used sources of hematopoietic stem cells for allogeneic transplants in pediatric practice are bone marrow (BM) and cord blood (CB). While bone marrow transplantation (BMT) is reaching its 30th year of application, human umbilical cord blood transplantation (HUCBT) is approaching its 10th. Although these procedures have basically the same purpose, a number of biological differences distinguish them. In particular, the intrinsically limited quantity of CB stem cells and their immunological naivete confer peculiar characteristics to these hematopoietic progenitors. From a bioethical point of view, the problems which have repeatedly been raised when the BM donor is a child are well-known. Different but no less important ethical problems are raised when one considers HUCBT; in this regard the most important issues are the easier propensity of programming a CB donor in comparison with a BM donor (clearly due to the shorter time interval needed to collect the hematopoietic progenitors); the in utero HLA typing; the implication of employing 'blood belonging to a neonate' for a third party; the need to perform a number of investigations both on the CB of the donor and on the mother and the implications that the discovery of disease may have for them, but also the need to establish banks for storing CB, with the accompanying administration and management problems. All these different aspects of UCBT will be discussed in the light of the four fundamental and traditional principles of bioethics, namely autonomy, nonmaleficence, beneficence and justice. PMID- 9208109 TI - Busulfan/cyclophosphamide in volunteer unrelated donor (VUD) BMT: excellent feasibility and low incidence of treatment-related toxicity. AB - An increasing number of volunteer unrelated donor bone marrow transplantations (VUD-BMT) are performed every year for hematological malignancies due to the availability of a large donor pool. Here we show the results of 36 VUD transplants from our institution using a chemotherapy-only conditioning regimen comprising busulfan 4 x 4 mg/kg and cyclophosphamide 2 x 60 mg/kg. All patients received heparin 200 IU/kg bw continuous i.v. infusion starting the day before conditioning until day +30. Thirty-four of 36 patients (94%) engrafted and no secondary graft failure was observed. The two non-engraftments occurred in patients with CML in blast crisis with extensive myelofibrosis. All 34 engrafted patients (100%) were in complete remission on day +30 as shown by bone marrow biopsy and cytogenetic examinations. No life-threatening treatment-related morbidity or mortality (TRM) were observed, in particular, no severe veno occlusive disease (VOD) of the liver and no fatal pulmonary complication. Use of G-CSF significantly shortened the time of neutropenia by 5 days. GVHD prophylaxis consisted of CsA/methylprednisolone with or without MTX. Acute GVHD grade II-IV was observed in 18/34 patients (53%) and cGVHD in 12/27 patients (45%), who survived to day +100. In seven patients (four with HLA class I or II mismatch) anti-T-lymphocyte globulin (ATG) was added for acute GVHD prophylaxis. One of seven had aGVHD grade II and none developed grade III to IV GVHD or graft failure. We conclude that Bu/CY is a feasible, save and sufficiently immunosuppressive regimen for VUD transplantation. Severe acute GVHD might be avoided by additional use of ATG in GVHD prophylaxis. PMID- 9208110 TI - GM-CSF-mobilized peripheral blood CD34+ cells differ from steady-state bone marrow CD34+ cells in adhesion molecule expression. AB - To determine the effect of growth factor mobilization on the expression of adhesion molecules, we compared CD34+ progenitor cell (PC) populations from steady-state bone marrow (BM) with granulocyte-macrophage colony-stimulating factor (GM-CSF)-mobilized apheresis products (peripheral blood stem cell (PSC)) using flow cytometry. To increase the accuracy of this analysis, CD34+ cells were enriched (MiniMACS) before cytometric analysis. A significantly lower expression of very late antigen-4 (VLA-4), leukocyte function antigen-1 (LFA-1) and LFA-3 were observed on PSC compared to BM CD34+ cells. In addition, significantly lower mean fluorescence intensity (MFI) of VLA-4, VLA-5, intercellular adhesion molecule-1 (ICAM-1), and sialyl Lewisx were observed on PSC as compared to BM CD34+ cells. Significantly higher levels of L-selectin and CD44 expression were observed on PSC as compared to BM CD34+ cells based on frequency and MFI (P < or = 0.05). In addition, the duration of GM-CSF administration or number of prior aphereses had no effect on adhesion molecule expression. These data suggest that decreased expression of adhesion molecules including VLA-4, LFA-1, ICAM-1 and LFA 3 play a role in PC mobilization. Based on these studies, we suggest that PC mobilization occurs as a stochastic process and is associated with the selection of CD34+ cells with low adhesion molecule expression. PMID- 9208111 TI - High-dose busulfan, melphalan, thiotepa and peripheral blood stem cell infusion for the treatment of metastatic breast cancer. AB - The purpose of this study was to determine the outcome of patients with metastatic breast cancer treated with high-dose busulfan (Bu), melphalan (Mel) and thiotepa (TT) followed by peripheral blood stem cell (PBSC) infusion. Fifty one patients with chemotherapy refractory (n = 32) or responsive (n = 19) metastatic breast cancer received Bu (12 mg/kg), Mel (100 mg/m2) and TT (500 mg/m2) followed by PBSC collected after chemotherapy and growth factor (n = 43) or growth factor alone (n = 8). The 100 day treatment-related mortality was 8% including one death from cytomegalovirus pneumonia, one from aspiration pneumonia and two from regimen-related toxicity (RRT). Seven of 28 refractory (25%) and 5/7 (71%) responsive patients with evaluable disease achieved a complete response of all measurable disease or all soft tissue disease with at least improvement in bone lesions (PR*). Fifteen of 51 patients (29%) are alive and progression-free a median of 423 days (range 353-934) after treatment, 5/32 (16%) with refractory disease and 10/19 (53%) with responsive disease. The probabilities of progression free survival (PFS) at 1.5 years for the patients with refractory (n = 32) and responsive (n = 19) disease were 0.24 and 0.53, respectively. These preliminary data suggest that high-dose Bu/Mel/TT has significant activity in patients with advanced breast cancer and may be superior to some previously published regimens. PMID- 9208112 TI - Excellent long-term survival after allogeneic marrow transplantation in patients with severe aplastic anemia. AB - Between 1982 and 1996, 20 patients (10 male, 10 female) with severe aplastic anemia (SAA) with a median age of 25 years (17-37 years), received grafts from an HLA-identical sibling (n = 17), HLA-identical unrelated donor (n = 2) or identical twin (n = 1). The median time from diagnosis to marrow transplantation (BMT) was 15 months (range 1-96 months). More than half of the patients had received more than 10 units of red blood cells or platelet transfusions prior to BMT. Pretransplant immunosuppression consisted of cyclophosphamide (CY) alone (n = 10), CY in combination with total body irradiation (n = 8), and CY and antithymocyte globulin (n = 2). For graft-versus-host disease (GVHD) prophylaxis methotrexate (MTX) alone (n = 9) or MTX with cyclosporin A (n = 10) were given. One patient died on day 18 after marrow grafting due to infection; all other patients had complete and sustained engraftment (95%). Eight patients developed acute GVHD (42%), nine patients chronic GVHD (53%) including four with extensive disease manifestation. One patient experienced a secondary malignancy 11 years after BMT. Eighteen patients followed for a median of 9.45 years (0.42-14.7 years) have sustained hematological reconstitution and are alive and well with a Karnofsky performance score of at least 90%. Thus, excellent long-term survival and low morbidity make allogeneic or syngeneic BMT the treatment of choice for younger patients with severe aplastic anemia. PMID- 9208113 TI - Haploidentical family member transplants for patients with chronic myeloid leukaemia: a report of the Chronic Leukaemia Working Party of off European Group for Blood and Marrow Transplantation (EBMT). AB - Within the registry of the chronic leukaemia working party of the European Group for Blood and Marrow Transplantation, data were available from 103 patients with chronic myeloid leukaemia who were treated by bone marrow transplantation from haploidentical family members. The patients of median age 30 years were transplanted between 1983 and 1994 in 25 European centres. The overall probabilities of survival and leukaemia-free survival (LFS) at 5 years were 32 and 25%, respectively. In univariate analysis, two factors were identified which affected survival and LFS, ie the state of disease at the time of transplant and the degree of HLA disparity. Fifty-nine patients were transplanted in first chronic phase and the probability of survival at 2 years was 47%. Forty-four patients received their transplants for advanced disease and their probability of survival at 2 years was 25% (P = 0.004). Donor bone marrow was HLA-mismatched for 0-1 antigens in 54 patients (group 1) and for 2-3 antigens in 49 patients (group 2). At 2 years, the probabilities of survival for groups 1 and 2 were 46 and 27% (P < 0.02) and the probabilities of LFS were 43 and 24% (P < 0.03), respectively. Multivariate analysis confirmed the prognostic importance of the disease stage and of the HLA disparity. Patients transplanted in first chronic phase from a donor mismatched for 0-1 HLA antigens had a probability of survival of 52% at 2 years compared with 19% for patients transplanted in advanced disease stage from donors mismatched for 2-3 HLA antigens. PMID- 9208114 TI - Survival in first or second remission after lymphocyte-depleted transplantation for Philadelphia chromosome-positive CML in first chronic phase. AB - We studied the outcome of BMT in 38 consecutive CML patients in CP1 who received transplants depleted of lymphocytes using counterflow centrifugation. In all patients the conditioning regimen was intensified by the addition of anthracyclines. Donors were HLA, MLC-identical siblings. Six patients (16%) died within 6 months. All 37 patients with a follow-up of more than 0.5 months engrafted and only one (3%) suffered from acute GVHD > or = grade 3. Chronic GVHD was evaluable in 33 patients and was extensive in six (18%). The projected 5-year probabilities of hematologic, cytogenetic and molecular relapse were 30% (95% confidence interval (CI), 10-49%), 35% (95% CI, 14-56%), and 34% (95% CI, 13 55%), respectively. The projected 5-year probability of survival was 68% (95% CI, 50-86%). Projected at 5 years, probabilities of leukemia-free survival (LFS) in hematologic, cytogenetic and molecular remission were 55% (95% CI, 37-73%), 51% (95% CI, 32-69%), and 51% (95% CI, 32-70%), respectively. All patients with relapse but one who relapsed in blastic phase were treated with retransplantation (n = 1) or with the infusion of lymphocytes (n = 6). Six patients regained second hematologic remission and five entered second cytogenetic and molecular remission. Including these patients, the probability of survival in first or second hematologic remission at the end of follow-up was 68% (95% CI, 50-86%). The probabilities of survival in first or second cytogenetic and molecular remission at the end of follow-up were both 61% (95% CI, 42-80%). We advocate revaluation of T cell depletion of donor marrow for patients with CML-CP1, especially for those at high risk of developing GVHD. PMID- 9208115 TI - Preferential sequestration in vitro of BCR/ABL negative hematopoietic progenitor cells among cytokine nonresponsive CML marrow CD34+ cells. AB - It is believed that long-term cultures of CML marrow cells favor the outgrowth of BCR/ABL negative hematopoietic progenitor cells (HPC) and that this phenomenon may be enhanced with negative hematopoietic regulators which can maintain primitive HPC in a quiescent state. Proliferation of CML marrow CD34+ cells in primary short-term cultures, maintained in the presence or absence of macrophage inhibitory protein-1 alpha (MIP-1 alpha), was tracked with the membrane dye PKH2. After 7 to 10 days it was possible to distinguish between cytokine responsive (CR) CD34+ cells (cells which had divided thus becoming PKH2dim) and cytokine nonresponsive (CNR) CD34+ cells (cells which had not divided and had therefore remained PKH2bright). CR and CNR CD34+ cells were isolated by flow cytometric cell sorting, seeded in secondary long-term cultures, and their progeny cells assayed weekly for their clonogenic progenitor cell content and expression of BCR/ABL by reverse transcriptase polymerase chain reaction (RT-PCR). Whereas CNR cells isolated from control primary cultures (control/CNR) sustained in vitro hematopoiesis, similar cells from cultures treated with MIP-1 alpha (MIP-1 alpha/CNR) supported a higher and, in some patients, a more extended production of clonogenic HPC, indicating that MIP-1 alpha was able to maintain primitive HPC in a quiescent state. Predominance of BCR/ABL negative progenitors in vitro was more evident in secondary cultures initiated with CNR cells than in those initiated with CR cells, especially those established with MIP-1 alpha/CNR cells. Of interest is the observed decline in the percentage of BCR/ABL+ progenitors in these cultures with time. Whereas up to 100% of progenitors were BCR/ABL+ on day 0, by day 14, only 46% of progenitors in MIP-1 alpha/CNR secondary cultures were BCR/ABL+ and by day 28 and beyond, the percentage of BCR/ABL+ progenitors dropped to below 20%. These results suggest that the quiescent nature of normal HPC present in CML marrow may favor their identification via cell tracking and, subsequently, their isolation from the more actively cycling leukemic cells. These studies also confirm the feasibility of employing negative hematopoietic regulators to augment the sequestration of normal HPC among the cytokine nonresponsive fraction of CD34+ cells, an approach that may be clinically feasible for autotransplantation. PMID- 9208117 TI - Liposomal amphotericin (AmBisome) is safe in bone marrow transplantation for primary immunodeficiency. AB - The use of conventional amphotericin B is limited by toxicity, side-effects, drug interactions and the need for large infusion volumes, especially for infants. Use of liposomal amphotericin B (AmBisome) in 15 paediatric BMT patients with primary immunodeficiency (PID) was therefore studied. Adverse clinical reactions to AmBisome and biochemical profiles were monitored daily for 2 weeks before, during and after each treatment episode. Fungal cultures were obtained weekly and when patients were pyrexial. There were 18 treatment episodes. Mean daily dose was 5 mg/kg (2-6 mg/kg). Mean duration of treatment was 25 days (5-90 days). Clinical reactions to AmBisome were observed in one infant who had a pyrexia of 38 degrees C. One of the 15 infants had a significant increase in creatinine level while on concomitant nephrotoxic therapy. Four developed mild hypokalaemia on AmBisome which resolved with increased potassium supplementation. AmBisome was well tolerated and without significant renal or hepatic toxicity in severely ill immunodeficient infants receiving multiple nephrotoxic and hepatotoxic drugs such as cyclosporin, vancomycin and foscarnet. PMID- 9208116 TI - Tumor cell contamination of peripheral blood stem cell transplants and bone marrow in high-risk breast cancer patients. AB - Twenty-one high-risk patients with primary stage II/III breast cancer were treated with high-dose chemotherapy comprising etoposide, ifosfamide, carboplatin and epirubicin (VIC-E). Tumor cells of epithelial origin were analyzed using the monoclonal antibodies CK2 (IgG1) and A45-B/B3 (IgG1) against cytokeratin (CK) components in bone marrow (BM) aspirates prior to chemotherapy, and in peripheral blood stem cell transplants (PBSCT). They were separated after the first (21/21 patients) and the second cycle (16/21 patients) of induction chemotherapy with VIP-E (etoposide, ifosfamide, cisplatin, epirubicin). Preliminary results showed CK positive tumor cells in 40% (14/35) of the analyzed transplants. In 7/12 (58.3%) patients, CK positive tumor cells were detectable in BM prior to treatment. Sixteen patients were separated after the 1st and 2nd cycle of VIP-E. PBSCT of 14/16 patients were assessable for presence of CK positive tumor cells. Our preliminary results demonstrate a lower tumor cell contamination of PBSCT separated after the 2nd cycle of induction therapy (14.3%) compared to contamination after the first induction therapy (64.3%). To date, 4/21 patients have experienced a relapse, and three of these patients had tumor cell positive transplants. Due to the small patient number only a trend towards a superior relapse-free survival in the patient group with CK negative transplants can be shown by Kaplan-Meier analysis. PMID- 9208118 TI - Tacrolimus and methotrexate for the prophylaxis of acute graft-versus-host disease in allogeneic bone marrow transplantation in patients with hematologic malignancies. AB - We conducted a study to evaluate the efficacy of the combination of tacrolimus and short-course methotrexate for the prevention of acute GVHD in patients with hematologic malignancies. Patients received preparative regimens specific for their disease category. Twenty-six out of 28 received HLA-identical sibling transplants and the two remaining patients received one-antigen mismatched transplants from a family member. With a median follow-up of 14 months, the Kaplan-Meier estimate of event-free survival was 50 +/- 9%. The probability of grade II-IV GVHD was 15 +/- 7%. Four patients developed GVHD: two had grade II and one each developed grade III and IV GVHD. Administration of methotrexate was associated with severe mucositis and there was no correlation between the distribution of the GVHD grade and the cumulative dose of methotrexate given. Thirteen patients have died; nine from transplant-related complications and four from relapse. The major toxicity of tacrolimus was renal. Nine out of 28 patients (32%) developed renal dysfunction attributed to tacrolimus. The combination of tacrolimus and methotrexate is an effective regimen for GVHD prophylaxis but associated with significant renal and mucosal toxicity. Further studies of tacrolimus as a single agent or in combination with either steroids or with a lower dose of methotrexate or with other antiproliferative drugs to modify the adverse events may improve the therapeutic index of this useful and promising agent. PMID- 9208120 TI - Prolonged remission of longstanding systemic lupus erythematosus after autologous bone marrow transplant for non-Hodgkin's lymphoma. AB - We describe a patient with longstanding steroid-dependent systemic lupus erythematosus (SLE) in whom clinical and serological remission was achieved following high-dose therapy and autologous bone marrow rescue for high-grade non Hodgkin's lymphoma. However, 3 years later, autoimmune disease re-presented in the form of immune thrombocytopenia (ITP), which had not previously been a feature of the SLE, necessitating reintroduction of steroid immunosuppression. Relapse of SLE is most likely, although de novo ITP post-BMT is also a possibility. The case suggests that severe long-standing autoimmune disease may be controlled by high-dose therapy and autologous stem cell reconstitution. However, further studies are required to determine the mechanism of re-emergence of autoimmunity and to evaluate optimal regimens and the potential value of such therapy in severe autoimmune diseases. PMID- 9208119 TI - The efficiency of tumor cell purging using immunomagnetic CD34+ cell separation systems. AB - Immunomagnetic separation with anti-CD34 monoclonal antibodies and paramagnetic microbeads has been used to enrich hematopoietic stem cells from human bone marrow (BM) or mobilized peripheral blood mononuclear cells (PBMNC). The introduction of this technique also constitutes a new principle of tumor cell purging. The efficiency in terms of purging tumor cells from PBMNC was evaluated in seven different experiments. Mobilized (chemotherapy and G-CSF) PBMNC were collected from patients with solid tumors (n = 6) and multiple myeloma (n = 1) by leukapheresis using an automated MNC separation system and contaminated with 1% (n = 5) or 10% (n = 2) tumor cells from different epithelial cell lines being CD34-negative. The cell mixture was sensitized with anti-CD34 (9C5) antibodies and sheep anti-mouse IgG1 paramagnetic microspheres and enriched for CD34+ cells using an Isolex 50 magnetic separator. Purify of CD34+ cells was studied by flow cytometry (FACScan) and tumor cell depletion was evaluated by comparative human tumor cloning assays (HTCA) containing methylcellulose and agar. We achieved a median purity of CD34+ cells of 85.9% (range 69.8-92.9%) and a median yield of 48.1% (range 21.0-85.2%). From these data in each case the estimated log depletion of tumor cells was calculated and compared with the experimentally achieved (HTCA) log depletion (log delta depletion = log experimental depletion- log calculated depletion). In our experiments we achieved a median depletion of 2.75 log (range 1.55-3.69 log). When corrected for CD34+ cell yield of each experiment we observed a median 'yield corrected depletion' of 2.38 log (range 1.48-3.15 log). The following delta depletion values were obtained: +0.32 log (HTB 129, breast), +0.21 log (HTB 26, breast), +0.04 log (HTB 26) for experiments with higher experimental depletion, and -0.23 log (HTB 26), -0.9 log (HTB 26, PBMNC from patient with multiple myeloma), -0.82 log (HTB 131, breast) and -1.66 log (HTB 131) for lower depletion efficacy than calculated. These data suggest that depletion may depend on specific cell surface characteristics of tumor cells. Moreover, plasma factors (eg paraprotein) may also have some impact. In summary, the Isolex 50 provides a high purity of CD34+ cells and depletion of tumor cells was efficient. However, calculated and experimental purging efficiencies are not necessarily identical. PMID- 9208121 TI - Bronchiolitis obliterans organizing pneumonia after syngeneic bone marrow transplantation for acute lymphoblastic leukemia. AB - We report a patient who developed bronchiolitis obliterans organizing pneumonia (BOOP) after syngeneic BMT for ALL. The patient complained of persistent low grade fever and non-productive cough after engraftment. Chest CT scan showed patchy infiltration bilaterally in the lower lung fields. Antibiotics were ineffective. Cultures, serological studies and polymerase chain reaction detected no infectious pathogens. We finally made a diagnosis of BOOP by thoracoscopical lung biopsy. The lung lesion disappeared in a month with corticosteroid therapy. While BOOP following allogeneic BMT has been reported, this is the first report after syngeneic transplantation. PMID- 9208122 TI - t(3;21) following peripheral blood stem cell transplantation in chronic phase chronic myeloid leukaemia. AB - A 40-year-old male with Ph-positive CML underwent PBSC autografting after initial treatment with hydroxyurea and interferon. Following autograft he remained in chronic phase with cytogenetic or molecular evidence of low levels of residual Ph positive cells. However, additional cytogenetic abnormalities, including t(3;21) typically seen in therapy-related myelodysplastic syndrome (MDS) and AML and blast crisis of CML, developed as an independent cell line following the autograft. More than 4 years after the autograft, the patient remains in chronic phase with no evidence of accelerated phase or blast crisis of CML, but with a concurrent MDS. We report a case of CML who developed therapy-related MDS following PBSC autograft while still remaining in chronic phase. PMID- 9208123 TI - Malaria infection after allogeneic bone marrow transplantation in a child with thalassemia. AB - A 12-year-old girl with beta-thalassemia hemoglobin E disease received a marrow transplant from her HLA-identical elder brother in July 1995. She had previously been treated by repeated blood transfusions. Conditioning included busulfan 16 mg/kg for 2 days and cyclophosphamide 120 mg/kg for 2 days. Cyclosporine was used for graft-versus-host disease prophylaxis. Spiking fevers occurred on days 6 and 11. Plasmodium falciparum parasites, both trophozoites and gametocytes, were found on the peripheral blood smear. Quinine 30 mg/kg three times a day for 7 days followed by a single dose of mefloquine 25 mg/kg was given. The fever subsided within 2 days and parasitemia cleared in 4 days. After transplant, the girl autologously reconstituted and was followed-up over 15 months. PMID- 9208125 TI - Inhibition by nociceptin of neurogenic inflammation and the release of SP and CGRP from sensory nerve terminals. AB - Pretreatment with the novel neuropeptide nociceptin (20 micrograms kg-1, i.p.) caused an inhibition of plasma extravasation evoked by antidromic stimulation of the saphenous nerve or by topical application of 1% mustard oil on the skin of the acutely denervated hindleg of the rat. In contrast, it did not affect non neurogenic inflammation evoked by s.c. injection of bradykinin after chronic denervation. Release of substance P (SP) and calcitonin gene-related peptide (CGRP) from rat isolated tracheae in response to electrical field stimulation was diminished by nociceptin (100 nM). It is concluded that nociceptin inhibits the release of sensory neuropeptides from terminals of nociceptive neurones. PMID- 9208124 TI - Cryptosporidiosis after CD34-selected autologous peripheral blood stem cell transplantation (PBSCT). Treatment with paromomycin, azithromycin and recombinant human interleukin-2. AB - We report two cases of cryptosporidiosis after CD34-selected PBSCT for lymphoma. While the first patient died of pulmonary cryptosporidiosis, treatment with paromomycin, azithromycin and subcutaneous low-dose rhIL-2 to improve numerical and functional T lymphocyte defects completely eliminated infection in the second patient. We conclude, that the removal of mature T lymphocytes by positive selection of CD34+ cells bears the risk of a delayed immune reconstitution resulting in an increased incidence of severe and sometimes fatal opportunistic infections. IL-2 might be useful in this situation by accelerating immune reconstitution and reducing the danger of opportunistic infections. PMID- 9208126 TI - Comparative direct electrophysiological effects of propofol on the conduction system and ionic channels of rabbit hearts. AB - 1. Propofol, an intravenous anaesthetic agent, can affect cardiac conduction but the ionic mechanisms have not been well defined. 2. This study measured the direct effects of propofol on the cardiac conduction system by using intracardiac recording/stimulation in Langendorff-perfused rabbit hearts. The underlying ionic mechanism was elucidated by using the whole-cell voltage clamp on rabbit isolated atrial and ventricular myocytes. 3. Propofol prolonged significantly the AV conduction (AH) interval at a clinically relevant concentration (3 microM). This AH interval prolongation was dose-dependent (3 to 100 microM). At higher concentrations, the AV nodal Wenckebach cycle length and its refractory period were also prolonged (10 to 100 microM). In addition, the conduction through the His-Purkinje system (HV interval) and the atrial tissue (SA interval), as well as the spontaneous cycle length, were lengthened dose-dependently (30 to 100 microM). 4. In isolated ventricular myocytes, Na current was decreased dose dependently by propofol. In part this was due to a negative-shift of the steady state voltage-dependent inactivation and a slowed rate of recovery from inactivation. The INa suppression by propofol was frequency-dependent. Propofol also blocked the ICa. The ED50 for peak current inhibition was 6.9 +/- 0.9 (n = 6) and 8.3 +/- 1.5 microM (n = 7) for INa and ICa, respectively. 5. The transient outward potassium current (Ito) of atrial myocytes was suppressed with an ED50 of 5.7 +/- 0.8 microM (n = 11), which was only partly caused by a left-shift of the steady-state inactivation. The inward rectifier K current (IK1) of the ventricular cells was reduced somewhat by propofol. 6. In summary, propofol can cause direct dromotropic and chronotropic effects on the cardiac conduction system, especially the atrioventricular node. These changes can be attributed, at least in part, to its direct dose-dependent suppression of the cardiac ICa, INa and Ito. Special concerns in the use of propofol anaesthesia for cardiac patients and the therapeutic antiarrythmic potential of propofol-like compounds are addressed. PMID- 9208127 TI - Attenuation of p53 expression and Bax down-regulation during phorbol ester mediated inhibition of apoptosis. AB - 1. Nitric oxide (NO) caused apoptotic cell death in murine RAW 264.7 macrophages. Associated with apoptotic morphology we observed p53 up-regulation and increased Bax expression. 12-O-tetradecanoylphorbol-13-acetate (TPA), a protein kinase C (PKC) activator potently blocked NO-induced apoptosis. To gain insights into the mechanisms involved we investigated the effect of TPA on apoptotic conveying proteins such as p53 and Bax. 2. TPA (100 nM) attentuated p53 up-regulation elicited by the NO-releasing compounds, S-nitrosoglutathione (1 mM) and sodium nitroprusside (1 mM), and suppressed p53 protein accumulation in response to endogenously generated NO. Hence, TPA appeared to lower the steady state p53 level following its up-regulation by NO. 3. Mezerein, a stage 2 tumour promoter and PKC activating agent was equally active to TPA. Moreover, two potent PKC inhibitors, staurosporine (10 nM) and Go 6976 (50 nM), reversed the inhibitory effect of TPA. However, bisinoylmaleimide I (up to 500 nM) was ineffective. 4. By extending the studies, we revealed a TPA-mediated p53 down-regulation in response to etoposide (50 microM), mitomycin C (5 micrograms ml-1) and actinomycin D (2 micrograms ml-1). 5. With the notion that TPA suppressed apoptotic DNA fragmentation in p53 antisense expressing cells as well, we searched for additional inhibitory actions of TPA. As well as affecting p53, TPA elicited a rapid decline of the steady state level of Bax within 30 min. 6. We concluded that down-regulation of two classical apoptotic promoting proteins contributes to the anti-apoptotic action of mezerein and TPA. PMID- 9208128 TI - Modulation by low sodium intake of glomerular response to cicletanine and atrial natriuretic factor. AB - 1. The aim of the study was to investigate whether cicletanine (CIC), as a potential inhibitor of cyclic GMP phosphodiesterase, is able to restore glomerular response to atrial natriuretic factor (ANF) in rats under conditions of diet deprived of sodium. We examined the effects of CIC on glomerular filtration rate (GFR), natriuresis and nephrogenous cyclic GMP excretion in response to ANF and the effects of both agents on intracapillary volume and cyclic GMP accumulation in isolated glomeruli of rats on normal and low sodium diets. 2. CIC (0.25 mg min-1 kg-1 BW) of ANF (0.5 microgram min-1 BW) alone, given in pharmacological doses, increased Cin significantly in normal sodium rats, whereas the effect of each agent was blunted in low sodium diet rats. Pretreatment with CIC restored the increase in C(in) in response to ANF infusion in low sodium diet rats. In rats on either diet, there were no differences in the extent of diuresis and natriuresis induced by CIC or ANF alone. In contrast to FENa, combined effects of both agents on V and UNa V in rats on normal and low sodium diets were observed. 3. In normal sodium diet rats, CIC 10(-4) M or ANF 10(-6) M alone inhibited angiotensin II 10(-6) M (AII)-induced decrease in intracapillary volume reflected by the glomerular [3H]-inulin space (GIS). In contrast, CIC or ANF alone did not inhibit AII-induced decrease in GIS in low sodium diet rats. Both agents given together inhibited AII-induced decrease in GIS in low sodium diet rats. 4. CIC both alone and in combination with ANF increased nephrogenous cyclic GMP excretion and cyclic GMP accumulation in isolated glomeruli of rats on normal and low sodium diets. In rats on either diet, CIC abolished the difference in ANF-stimulated increase in nephrogenous cyclic GMP excretion and cyclic GMP accumulation in glomeruli. 5. These results suggest that CIC and ANF alone induce relaxation of glomeruli and a resultant increase in glomerular filtration rate in normal sodium diet rats; in contrast, these effects are blunted in the low sodium diet rats. CIC restores glomerular response to ANF in low sodium diet rats, apparently involving inhibition of the cyclic GMP phosphodiesterase. PMID- 9208129 TI - The stimulation of human neutrophil migration by angiotensin IL: its dependence on Ca2+ and the involvement of cyclic GMP. AB - 1. Angiotensin II had a bimodal effect on human neutrophil migration. Low concentrations of angiotensin II stimulated random migration. At a concentration of 10(-10) M it caused a maximal increase of migration; migration increased from 47.2 +/- 2.1 microns in the absence of angiotensin II, to 73.1 +/- 2.2 microns with 10(-10) M angiotensin II present in the lower compartment of the Boyden chamber (n = 5, P < 0.001). Stimulation of migration by angiotensin II was partly chemotactic and partly chemokinetic. Angiotensin II concentrations of 10(-8) M and higher inhibited chemotactic peptide-stimulated chemotaxis. 2. The stimulant effect of angiotensin II on migration was completely dependent on extracellular Ca2+. In the presence of 1 mM Ca2+, angiotensin II stimulated migration to 76.1 +/- 1.7 microns, while migration in the absence of Ca2+ was 42.2 +/- 1.9 microns (n = 4, P < 0.001). Different types of calcium channel blockers either moderately or strongly inhibited angiotensin II-activated migration. Stimulation of migration by angiotensin II in intact cells required higher concentrations of Ca2+ than in electroporated cells. This supports the view that there is an influx of Ca2+ through the plasma membrane, and a requirement of calcium for an intracellular target. 3. Angiotensin II-stimulated migration was inhibited by pertussis toxin; from 71.6 +/- 2.0 microns in the absence, to 43.6 +/- 1.5 microns in the presence of pertussis toxin (n = 4, P < 0.001). Migration of electroporated neutrophils stimulated by angiotensin II was synergistically enhanced by GTP gamma S. This suggests that one or more G-proteins are involved in the activating effect of angiotensin II. 4. Inhibitors of soluble guanylate cyclase and antagonists of cyclic GMP-dependent kinase strongly inhibited the activating effect of angiotensin II. The results suggest that the activating effect of angiotensin II is mediated by cyclic GMP and by cyclic GMP-dependent kinase. PMID- 9208130 TI - The expression of functionally-coupled B2-bradykinin receptors in human corneal epithelial cells and their pharmacological characterization with agonists and antagonists. AB - 1. Bradykinin (BK) and Lys-BK are peptides which are released at high nanomolar concentrations into the tear-film of ocular allergic patients. We hypothesized that these peptides may activate specific receptors on the ocular surface, especially the corneal epithelium (CE) and thus the CE cells may represent a potential target tissue for these kinins. 2. The purpose of the present studies, therefore, was to determine the presence of and the pharmacological characteristics of bradykinin receptors on normal cultured primary and SV40 virus transformed human corneal epithelial (CEPI) cells by use of the accumulation of [3H]-inositol phosphates ([3H]-IPs) as a bioassay. 3. Bradykinin (BK) induced a maximal 1.95 +/- 0.24 fold (n = 17) and 2.51 +/- 0.29 fold (n = 26) stimulation of [3H]-IPs accumulation in normal, primary (P-CEPI) and SV40-immortalized (CEPI 17-CL4) cells, respectively. This contrasted with a maximal 3.2-4.5 fold and 2.0 2.9 fold stimulation by histamine (100 microM) and platelet activating factor (100 nM) in both cell-types, respectively. 4. The molar potencies of BK and some of its analogues in the CEPI-17-CL4 cells were as follows: BK (EC50 = 3.26 +/- 0.61 nM, n = 18), Lys-BK (EC50 = 0.95 +/- 0.16 nM, n = 5), Met-Lys-BK (EC50 = 2.3 +/- 0.42 nM, n = 5), Ile-Ser-BK (EC50 = 5.19 +/- 1.23 nM, n = 6), Ala3-Lys-BK (EC50 = 12.7 +/- 2.08 nM, n = 3), Tyr8-BK (EC50 = 19.3 +/- 0.77 nM, n = 3), Tyr5 BK (EC50 = 467 +/- 53 nM, n = 4) and des-Arg9-BK (EC50 = 14.1 +/- 2.7 microM, n = 4). The potencies of BK-related peptides in normal, P-CEPI cells were similar to those found in transformed cells, thus: BK, EC50 = 2.02 +/- 0.69 nM (n = 7), Tyr8 BK, EC50 = 14.6 +/- 2.7 nM (n = 3), Tyr5 = BK, EC50 = 310 +/- 70 nM (n = 4) and des-Arg9-BK, EC50 = 12.3 +/- 3.8 microM (n = 3). 5. The bradykinin-induced responses were competitively antagonized by the B2-receptor selective BK antagonists, Hoe-140 (D-Arg-[Hyp3, Thi5, D-Tic7, Oic8]BK; Icatibant; molar antagonist potency = 2.9 nM; pA2 = 8.54 +/- 0.06, n = 4; and slope = 1.04 +/- 0.08) and D-Arg0[Hyp3,Thi5,8, DPhe7]-BK (KB = 371 nM; pKB = 6.43 +/- 0.08, n = 4) in CEPI-17-CL4 cells. The antagonist potency of Hoe-140 against BK in normal, P CEPI cells was 8.4 +/- 1.8 nM (pKi = 8.11 +/- 0.12, n = 4), this being similar to the potency observed in the immortalized cells. 6. This rank order of potency of agonist BK-related peptides, coupled with the antagonism of the BK-induced [3H] IPs by the specific B2-receptor antagonists, strongly suggests that a B2-receptor subtype is involved in mediating functional phosphoinositide (PI) responses in the CEPI-17-CL4 and P-CEPI cells. 7. In conclusion, these data indicate that the P-CEPI and CEPI-17-CL4 cells express BK receptors of the B2-subtype coupled to the PI turnover signal transduction pathway. The CEPI-17-CL4 cells represent a good in vitro model of the human corneal epithelium in which to study further the role of BK receptors in its physiology and pathology, such as in allergic/inflammatory conditions, potential wound healing and other functions of the cornea. PMID- 9208132 TI - The involvement of reactive oxygen species and arachidonic acid in alpha 1 adrenoceptor-induced smooth muscle cell proliferation and migration. AB - 1. In a previous study, we demonstrated phenylephrine-stimulated arachidonic acid (AA) release in rabbit cultured aortic smooth muscle cells. Therefore, we have investigated the functional implications of AA which are involved in the cellular response to phenylephrine, particularly proliferation and migration of rabbit cultured aortic smooth muscle cells. 2. First, to determine whether AA directly modifies proliferation and mobility of vascular smooth muscle cells (VSMCs), we exposed the cells to AA. AA induced proliferation and migration of the cells in a dose-dependent fashion. Concomitantly added catalase inhibited the proliferation and chemotaxis induced by AA of VSMCs. Conversely, aminotriazole enhanced the proliferation and migration induced by AA. 3. Secondly, we investigated whether the proliferation and migration of VSMCs by phenylephrine were related to AA and hydrogen peroxide (H2O2). The proliferation and chemotaxis of VSMCs by phenylephrine were inhibited by a phospholipase A2 (PLA2) inhibitor, or catalase. 4. Lastly, we investigated the effects of AA and phenylephrine on the content of H2O2 in VSMCs. AA and phenylephrine treatment led to an increase of H2O2 in a dose-dependent manner. 5. These results suggest that the addition of phenylephrine to the cells caused the enhancement of proliferation and migration, probably by mediating AA release and reactive oxygen species (ROS) production. PMID- 9208131 TI - Characterization of endothelium-derived relaxing factors released by bradykinin in human resistance arteries. AB - 1. Relaxing factors released by the endothelium and their relative contribution to the endothelium-dependent relaxation produced by bradykinin (BK) in comparison with different vasodilator agents were investigated in human omental resistance arteries. 2. BK produced an endothelium-dependent relaxation of arteries pre contracted with the thromboxane A2 agonist, U46619. The B2 receptor antagonist, Hoe 140 (0.1, 1 and 10 microM), produced a parallel shift to the right of the concentration-response curve to BK with a pA2 of 7.75. 3. Neither the cyclo oxygenase inhibitor, indomethacin (10 microM) alone, the nitric oxide synthase inhibitor, N omega-nitro-L-arginine methyl ester (L-NAME, 300 microM) alone, the nitric oxide scavenger, oxyhaemoglobin (Hb, 10 microM) alone, nor the combination of L-NAME plus Hb affected the concentration-response curve to BK. Conversely, the combination of indomethacin with either L-NAME or Hb attenuated but did not abolish the BK-induced relaxation. By contrast, the relaxations produced by the Ca2+ ionophore, calcimycin (A23187), and by the inhibitor of sarcoplasmic reticulum Ca(2+)-ATPase, thapsigargin (THAPS), were abolished in the presence of indomethacin plus L-NAME. Also, the presence of indomethacin plus L-NAME produced contraction of arteries with functional endothelium. 4. The indomethacin plus L NAME resistant component of BK relaxation was abolished in physiological solution (PSS) containing 40 mM KCl and vice versa. However, in the presence of KCl 40 mM, indomethacin plus L-NAME did not affect the nitric oxide donor, S-N acetylpenicillamine-induced relaxation. 5. The indomethacin plus L-NAME resistant component of the relaxation to BK was significantly attenuated by the K+ channel blocker tetrabutylammonium (TBA, 1 mM). However, it was not affected by other K+ channel blockers such as apamin (10 microM), 4-aminopyridine (100 microM), glibenclamide (10 microM), tetraethylammonium (10 mM) and charybdotoxin (50 nM). 6. In the presence of indomethacin plus L-NAME, the relaxation produced by BK was not affected by the phospholipase A2 inhibitor, quinacrine (10 microM) or by the inhibitor of cytochrome P450, SKF 525a (10 microM). Another cytochrome P450 inhibitor, clotrimazole (10 microM) which also inhibits K+ channels, inhibited the relaxation to BK. 7. These results show that BK induces endothelium-dependent relaxation in human small omental arteries via multiple mechanisms involving nitric oxide, cyclo-oxygenase derived prostanoid(s) and another factor (probably an endothelium-derived hyperpolarizing factor). They indicate that nitric oxide and cyclo-oxygenase derivative(s) can substitute for each other in producing relaxation and that the third component is not a metabolite of arachidonic acid, formed through the cytochrome P-450 pathway, in these arteries. PMID- 9208133 TI - The effect of R 15.7/HO, an anti-CD18 antibody, on the late airway response and airway hyperresponsiveness in an allergic rabbit model. AB - 1. The effects of a mouse (IgG1 fraction) anti-CD 18 neutralizing antibody (R15.7) on allergen-induced late airway response (LAR), airway hyperresponsiveness (AHR) and cellular recruitment were investigated in an allergic rabbit model. 2. Litter-matched NZW rabbits immunized within 24 h of birth with Alternaria tenuis (i.p.) and subsequently exposed to the allergen (i.p.) for the first 3 months of life were challenged with inhaled allergen as adult rabbits. Lung function in terms of dynamic compliance (Cdyn; ml cmH2O-1) and total lung resistance (RL; cmH2O-1 s-1) was monitored for 6 h following the allergen challenge. On day 16, separate groups of rabbits were pretreated with either control antibody (a non-binding mouse IgG1, 1 mg kg-1, i.v.) or R15.7 (1 mg kg-1, i.v.) and 1 h later all were challenged with Alternaria tenuis and lung function monitored thereafter. Airway responsiveness to inhaled histamine was assessed by measuring RL and Cdyn 24 h before and after allergen challenge and bronchoalveolar lavage (BAL) was also performed 24 h before and after allergen challenge. 3. Pretreatment of rabbits with the control antibody had no effect on the LAR as measured by AUC (Cdyn, 0-6 h). However, the magnitude of the LAR following treatment with R15.7 was significantly reduced when compared to LAR demonstrated on 1st challenge (P < 0.001) or to that of the control group on both challenges (P < 0.01). 4. In control antibody pretreated rabbits allergen induced a significant 3.4 fold reduction in the PC50 response to inhaled histamine in terms of RL changes (P < 0.05) and a significant 2.1 fold reduction in PC35 response to inhaled histamine in terms of Cdyn changes (P < 0.05). However, in anti-CD 18 antibody pretreated rabbits there was no significant change in responsiveness to histamine 24 h following allergen, as assessed by either RL PC50 or Cdyn PC35. 5. Allergen challenge induced a significant increase in eosinophil and neutrophil numbers (P < 0.05) in rabbits pre-treated with control antibody, whereas treatment with R15.7 significantly inhibited this increase in the numbers of both cell types. 6. This study demonstrates that the neutralization of CD-18 molecules reduces allergen-induced infiltration of both eosinophils and neutrophils into the airways and abolishes the accompanying LAR and AHR. These results provide evidence to support a role for CD-18 adhesion molecules in the transmigration of inflammatory cells into airways. PMID- 9208134 TI - Vasodilator effects of adrenomedullin on small pulmonary arteries and veins in anaesthetized cats. AB - 1. This study was conducted to determine adrenomedullin (AM) action sites in the pulmonary vascular bed and the relation between its vasodilator effects and vascular tone. Moreover, an examination was made into whether calcitonin gene related peptide (CGRP) receptors mediate pulmonary vasodilatations induced by AM. To this end, we directly measured internal diameter (i.d.) changes in small pulmonary arteries and veins (100-1100 microns i.d.) by use of an X-ray television system on the in vivo cat lung. 2. Under control (resting vascular tone) conditions, AM injections into the left main pulmonary artery caused dose related i.d. increases in both small arteries and veins. The mean i.d. increase of the 100-1100 microns arteries (4 +/- 1, 11 +/- 2, and 17 +/- 2% with 0.01, 0.1, and 1 nmol kg-1 AM, respectively) was significantly larger than that for the veins (1 +/- 1, 5 +/- 2, and 7 +/- 2% with 0.01, 0.1 and 1 nmol kg-1 AM, respectively) whatever the injected dose of AM. 3. When unilobar hypoxia (5% O2) had decreased the i.d. of the 100-1100 microns arteries and veins by 16 +/- 3 and 6 +/- 3%, respectively, AM (0.1 nmol kg-1) was able to induce significantly larger i.d. increases in the arteries (28 +/- 3%) and veins (11 +/- 3%) than those under control conditions. 4. The AM-induced i.d. response pattern in the serially connected pulmonary arteries was quite different from that induced by CGRP; AM caused a greater increase in smaller vessels (100-500 microns) than in larger vessels (500-1100 microns). In the case of CGRP, a greater increase was observed in the larger vessels. 5. CGRP8-37 (100 nmol kg-1, i.v., followed by a continuous infusion of 0.2 nmol kg-1 min-1) had no significant effect on the i.d. increase induced by AM (0.1 nmol kg-1) in any serial segments of the arteries and veins. 6. The results indicate that, in the cat, AM induces greater vasodilatation in small pulmonary arteries and lesser vasodilatation in small veins, the maximum dilatation being in the more peripheral arterial segment (100 500 microns). The vasodilator effect of AM was enhanced when vascular tone was elevated. The data suggest that the AM-induced pulmonary vasodilatation is not mediated by CGRP receptors but by its own specific receptor. PMID- 9208135 TI - Endothelin receptors and their cellular signal transduction mechanism in human cultured prostatic smooth muscle cells. AB - 1. Endothelin (ET) receptors, and their cellular signal transduction mechanism, were characterized in a primary culture of human prostatic smooth muscle cells (HP cell). 2. [125I]-ET-1 and [125I]-ET-3 binding studies revealed that both ETA and ETB receptors were present in the HP cells, and the ratio of ETA to ETB receptors was 1.4:1. 3. Analysis of ET receptor mRNA by reverse transcription polymerase chain reaction also demonstrated that HP cells express both ETA and ETB receptors. 4. ET-1 and ET-3 increased intracellular free Ca2+ concentration ([Ca2+]i) in the HP cells in a concentration-dependent manner. Use of subtype selective antagonists BQ-123 and BQ-788, indicated that both ETA and ETB receptors were coupled to an increase in [Ca2+]i. 5. Pretreatment of the cells with pertussis toxin resulted in a significant but partial attenuation of the [Ca2+]i increase mediated through the ETA and ETB receptors. However, sensitivity to pertussis toxin (PTX) was significantly different between them. 6. In conclusion, HP cells possess ETA and ETB receptors. Further, these two endothelin receptor subtypes evoke an increase in [Ca2+]i possibly via the action of different GTP-binding proteins. PMID- 9208136 TI - Effect of calpain inhibitor I, an inhibitor of the proteolysis of I kappa B, on the circulatory failure and multiple organ dysfunction caused by endotoxin in the rat. AB - 1. We compared the effects of calpain inhibitor I (inhibitor of the proteolysis of I kappa B and, hence, of the activation of nuclear factor kappa B (NF kappa B) and dexamethasone on (i) the circulatory failure, (ii) multiple organ dysfunction and (iii) induction of the inducible isoforms of nitric oxide (NO) synthase (iNOS) and cyclo-oxygenase (COX-2) in anaesthetized rats with endotoxic shock. 2. Injection of lipopolysaccharide (LPS, E. coli, 10 mg kg-1, i.v.) resulted in hypotension and a reduction of the pressor responses elicited by noradrenaline. This circulatory dysfunction was attenuated by pretreatment of LPS-rats with calpain inhibitor I (10 mg kg-1, i.v., 2 h before LPS) or dexamethasone (1 mg kg 1, i.v.). 3. Endotoxaemia also caused rises in the serum levels of (i) urea and creatinine (renal dysfunction), (ii) alanine aminotransferase (ALT), aspartate aminotransferase (AST) (hepatocellular injury), bilirubin and gamma-glutamyl transferase (gamma GT) (liver dysfunction), (iii) lipase (pancreatic injury) and (iv) lactate. Calpain inhibitor I and dexamethasone attenuated the liver injury, the pancreatic injury, the lactic acidosis as well as the hypoglycaemia caused by LPS. Dexamethasone, but not calpain inhibitor I, reduced the renal dysfunction caused by LPS. 4. Endotoxaemia for 6 h resulted in a substantial increase in iNOS and COX-2 protein and activity in lung and liver, which was attenuated in LPS rats pretreated with calpain inhibitor I or dexamethasone. 5. Thus, calpain inhibitor I and dexamethasone attenuate (i) the circulatory failure, (ii) the multiple organ dysfunction (liver and pancreatic dysfunction/injury, lactic acidosis, hypoglycaemia), as well as (iii) the induction of iNOS and COX-2 protein and activity in rats with endotoxic shock. We propose that prevention of the activation of NF-kappa B in vivo may be useful in the therapy of circulatory shock or of disorders associated with local or systemic inflammation. PMID- 9208137 TI - Effect of a novel selective and potent phosphinic peptide inhibitor of endopeptidase 3.4.24.16 on neurotensin-induced analgesia and neuronal inactivation. AB - 1. We have examined a series of novel phosphinic peptides as putative potent and selective inhibitors of endopeptidase 3.4.24.16. 2. The most selective inhibitor, Pro-Phe-psi(PO2CH2)-Leu-Pro-NH2 displayed a Ki value of 12 nM towards endopeptidase 3.4.24.16 and was 5540 fold less potent on its related peptidase endopeptidase 3.4.24.15. Furthermore, this inhibitor was 12.5 less potent on angiotensin-converting enzyme and was unable to block endopeptidase 3.4.24.11, aminopeptidases B and M, dipeptidylaminopeptidase IV and proline endopeptidase. 3. The effect of Pro-Phe-psi(PO2CH2)-Leu-Pro-NH2, in vitro and in vivo, on neurotensin metabolism in the central nervous system was examined. 4. Pro-Phe psi(PO2CHH2)-Leu-Pro-NH2 dose-dependently inhibited the formation of neurotensin 1-10 and concomittantly protected neurotensin from degradation by primary cultured neurones from mouse embryos. 5. Intracerebroventricular administration of Pro-Phe-psi(PO2CH2)-Leu-Pro-NH2 significantly potentiated the neurotensin induced antinociception of mice in the hot plate test. 6. Altogether, our study has established Pro-Phe-psi(PO2CH2)-Leu-Pro-NH2 as a fully selective and highly potent inhibitor of endopeptidase 3.4.24.16 and demonstrates, for the first time, the contribution of this enzyme in the central metabolism of neurotensin. PMID- 9208138 TI - Effects of calcium dobesilate on the synthesis of endothelium-dependent relaxing factors in rabbit isolated aorta. AB - 1. Some cardiovascular disturbances which occur in diabetics are a consequence of alterations in vascular contractility as well as in endothelium-dependent relaxation. 2. Calcium dobesilate (DOBE) is a drug used in diabetic retinopathy and its mechanism of action is not yet understood. 3. The aim of this study was to investigate the effects of DOBE on synthesis and release of endothelium dependent relaxing factor (EDRF) and endothelium-dependent hyperpolarizing factor (EDHF) in rabbit isolated aorta. 4. Endothelium-dependent relaxation induced by acetylcholine (ACh) (10(-8)-(10(-5) M) increased in the presence of DOBE 10(-5) M only when vascular endothelium was kept intact. 5. NG-nitro-L-arginine methyl ester (L-NAME; 10(-8)-10(-4) M progressively decreased the enhancing effect of DOBE on endothelium-dependent relaxation whereas it was progressively increased by L-Arg. 6. DOBE 10(-5) M increased in a non-significant manner endothelium dependent relaxation induced by ACh when the arteries were incubated with both L NAME 10(-4) M and indomethacin 10(-5) M. 7. DOBE (10(-6) M and 10(-5) M) was able to scavenge superoxide anion radicals generated by the hypoxanthine/xanthine oxidase reaction. 8. These results provide evidence that DOBE is able to affect the vascular disorders associated with diabetes mellitus since it enhances the synthesis of endothelium-dependent relaxing factors. PMID- 9208139 TI - Protection by dexamethasone of the functional desensitization to beta 2 adrenoceptor-mediated responses in human lung mast cells. AB - 1. The beta-adrenoceptor agonist, isoprenaline, inhibited the IgE-mediated release of histamine from human lung mast cells (HLMC) in a dose-dependent manner. Maximal inhibitory effects were obtained with 0.1 microM isoprenaline. However, the inhibition of histamine release from HLMC by isoprenaline (0.1 microM) was highly variable ranging from 33 to 97% inhibition (mean, 59 +/- 3%, n = 27). 2. Long-term (24 h) incubation of HLMC with isoprenaline led to a subsequent reduction in the ability of a second exposure of isoprenaline to inhibit IgE-mediated histamine release from HLMC. The impairment in the ability of isoprenaline (0.1 microM) to inhibit histamine release following desensitizing conditions (1 microM isoprenaline for 24 h) was highly variable amongst HLMC preparations ranging from essentially negligible levels of desensitization in some preparations to complete abrogation of the inhibitory response in others (mean, 65 +/- 6% desensitization, n = 27). 3. The ability of HLMC to recover from desensitization was investigated. Following desensitizing conditions (1 microM isoprenaline for 24 h), HLMC were washed and incubated for 24 h in buffer and the effectiveness of isoprenaline (0.1 microM) to inhibit IgE-mediated histamine release from HLMC was assessed. The extent of recovery was highly variable with some HLMC preparations failing to recover and others displaying a complete restoration of responsiveness to isoprenaline (mean, 40 +/- 6% recovery, n = 23). 4. The effects of the glucocorticoid, dexamethasone, were also investigated. Long term (24-72 h) treatments with dexamethasone (0.1 microM) had no effect on IgE mediated histamine release from HLMC. Additionally, long-term (24-72 h) treatments with dexamethasone (0.1 microM) had no effect on the effectiveness of isoprenaline to inhibit histamine release. However, long-term (24-72 h) treatments with dexamethasone (0.1 microM) protected against the functional desensitization induced by incubation (24 h) of HLMC with isoprenaline (1 microM). The protective effect was time-dependent and pretreatment of HLMC with dexamethasone for either 24, 48 or 72 h prevented desensitization by either 15 +/ 7, 19 +/- 5 or 51 +/- 10%, respectively (n = 5-7). 5. HLMC preparations which were relatively refractory to isoprenaline even after withdrawal (24 h) from desensitizing conditions responded more effectively to isoprenaline (0.1 microM) if dexamethasone (0.1 microM) was also included during the recovery period (19 +/ 9% recovery after 24 h in buffer; 50 +/- 8% recovery after 24 h with dexamethasone, n = 5). 6. These data indicate that the responses of different HLMC preparations to isoprenaline, the susceptibility of HLMC to desensitization and the ability of HLMC to recover from desensitizing conditions varies markedly. Dexamethasone, which itself has no direct effects on IgE-mediated histamine release from HLMC, protected HLMC from the functional desensitization to beta adrenoceptor agonists. Because beta 2-adrenoceptor agonists and glucocorticoids are important in the therapeutic management of asthma and as the HLMC is probably important in certain types of asthma, these findings may have wider clinical implications. PMID- 9208140 TI - Effects of an orally active non-peptide bradykinin B2 receptor antagonist, FR173657, on plasma exudation in rat carrageenin-induced pleurisy. AB - 1. Effects of an orally active non-peptide (BK) B2 receptor antagonist, FR173657 ((E)-3-(6-acetamido-3-pyridyl)-N-[N-[2,4-dichloro-3-[(2-methyl-8-quinoli nyl) oxymethyl]phenyl]-N-methylaminocarbonylmethyl] acrylamide) on the plasma exudation in rat carrageenin-induced pleurisy were investigated. 2. Plasma exudation induced by intrapleural injection of bradykinin (BK, 3 nmol per rat) into male SD strain rats (SPF, 8 weeks old) were significantly inhibited by oral administration of novel B2 receptor antagonist FR173657 (3-30 mg kg-1, 1 h before BK injection) in a dose-dependent manner, whereas that induced by histamine was not. 3. The inhibitory effect of 30 mg kg-1 FR173657 persisted for more than 4 h. 4. Intrapleural injection of lambda-carrageenin (2% (w/v), 0.1 ml per rat) caused marked plasma exudation and accumulation of exudates from 1 h after carrageenin injection. The maximum plasma exudation response was observed 5 h after carrageenin. The oral administration of FR173657 to rats (30 mg kg-1, 1 h before carrageenin) significantly (by 50-77%) blunted the plasma exudation 1, 3, 5, and 7 h after carrageenin, causing a significant parallel reduction (by 42-57%) in the volume of exudates. 5. The anti-inflammatory effect of FR173657 on rat carrageenin-induced pleurisy was almost equipotent with that of the peptide B2 antagonist Hoe140 (1 mg kg-1, i.v.), a plasma kallikrein inhibitor, soy bean trypsin inhibitor (0.3 mg per rat, intrapleural injection) and bromelain (10 mg kg-1, i.v.). 6. In pleurisy induced by intrapleural injection of a histamine releaser, compound 48/80, the plasma exudation was observed only within 20 min after the injection. This plasma exudation was not affected by FR173657, although it was completely inhibited by a mixture of pyrilamine (5 mg kg-1, i.v.) and methysergide (3 mg kg-1, i.v.). 7. These results indicate that FR173657 is an orally active, promising anti-inflammatory agent for kinin-dependent inflammation. PMID- 9208141 TI - Evidence that antipsychotic drugs are inverse agonists at D2 dopamine receptors. AB - 1. The effects of a number of D2-like dopamine receptor antagonists have been determined on forskolin-stimulated cyclic AMP accumulation in Chinese hamster ovary (CHO) cells expressing the human D2short dopamine receptor (CHO-D2S cells). 2. Dopamine inhibited the effect of forskolin (as expected for a D2 receptor). However, all of the antagonists tested, apart from UH232 and (-)-butaclamol, were able to increase cyclic AMP accumulation above the forskolin control level. (+) Butaclamol elicited a similar stimulation of forskolin-stimulated cyclic AMP accumulation in a CHO cell line expressing human D2long dopamine receptors whereas it exhibited no stimulating effect on forskolin-stimulated cyclic AMP accumulation in untransfected CHO-K1 cells. 3. There was a strong correlation between the EC50 values of these compounds for potentiation of cyclic AMP accumulation and their Ki values from radioligand binding experiments in CHO-D2S cells. 4. The effects of both (+)-butaclamol and dopamine in CHO-D2S cells were inhibited by pre-treatment with pertussis toxin indicating a role for Gi/Go proteins. 5. UH232 did not significantly affect forskolin-stimulated cyclic AMP accumulation but this substance was able to inhibit the effects of both dopamine and (+)-butaclamol in a concentration-dependent manner. Thus the effects of (+) butaclamol on forskolin-stimulated cyclic AMP accumulation are mediated directly via the D2 receptor rather than by reversal of the effects of an endogenous agonist. 6. These data suggest that the D2 dopamine receptor antagonists tested here, many of which are used clinically as antipsychotic drugs, are in fact inverse agonists at human D2 dopamine receptors. PMID- 9208142 TI - (+)-WAY 100135, a partial agonist, at native and recombinant 5-HT1B/1D receptors. AB - 1. We have studied the effects of the purportedly selective 5-HT1A receptor antagonist (+)-WAY 100135 on electrically stimulated 5-hydroxytryptamine (5-HT) efflux in the ventrolateral geniculate nucleus (vLGN), and its affinity at human 5-HT1B and 5-HT1D receptors stably expressed in Chinese hamster ovary (CHO) cells. 2. On short 'pseudo single pulse' stimulations (20 pulses at 100 Hz, 190 ms train duration), (+)-WAY 100135 (1.0 microM) decreased 5-HT efflux in the vLGN to 68 +/- 8% of pre-drug values (P < 0.01). This decrease could be blocked by the 5-HT1D/1B receptor antagonist GR 127935 (50 nM). Conversely, when long stimulations (20 pulses at 20 Hz, 950 ms train) were used, (+)-WAY 100135 had no effect on 5-HT efflux (84 +/- 8% of pre-drug values) although both methiothepin (200 nM) and GR 127935 (50 nM) caused significant increases (to 175 +/- 18 and 130 +/- 10% of pre-drug values, respectively). 3. Paroxetine (100 nM), the selective 5-HT reuptake inhibitor, increased stimulated 5-HT efflux and reuptake half-life (to 145 +/- 18% and 649 +/- 121%, respectively) on pseudo single pulse stimulations. When (+)-WAY 100135 was added in combination with the uptake blocker, the effect of paroxetine on stimulated 5-HT efflux was potentiated to 282 +/- 48% (P < 0.01) without further effect on the 5-HT reuptake half-life. 4. The affinity and intrinsic activity of (+)-WAY 100135 were determined at recombinant human 5-HT1B and 5-HT1D receptors expressed in CHO cells, by use of radioligand binding and [35S]-GTP gamma S binding (+)-WAY 100135 was a partial agonist at human 5-HT1B and 5-HT1D receptors with moderately high affinity for 5 HT1D receptors (pEC50 = 7.61). 5. In conclusion, (+)-WAY 100135 was found to be not a selective 5-HT1A autoreceptor antagonist but may act as a partial agonist at the 5-HT1B/1D receptor, displaying agonist or antagonist properties depending on the stimulation protocol used and the resultant 5-HT 'tone' at the receptor. PMID- 9208143 TI - Evidence that cyclic AMP phosphodiesterase inhibitors suppress interleukin-2 release from murine splenocytes by interacting with a 'low-affinity' phosphodiesterase 4 conformer. AB - 1. We have investigated the suppressive effects of rolipram, RP 73401 (piclamilast) and other structurally diverse inhibitors of cyclic AMP-specific phosphodiesterase 4 (PDE4) on interleukin (IL)-2 generation from Balb/c mouse splenocytes exposed to the superantigen, Staphylococcocal enterotoxin-A (Staph. A). The purpose was to determine whether their potencies are more closely correlated with inhibition of PDE4 from CTLL cells, against which rolipram displays weak potency (low-affinity PDE4), or displacement of [3H]-(+/-)-rolipram from its high-affinity binding site (HARBS) in mouse brain cytosol. 2. RP 73401 (IC50 0.46 +/- 0.07 nM, n = 4) was a very potent inhibitor of Staph. A-induced IL 2 release from Balb/c mouse splenocytes, being > 1100 fold more potent than (+/-) rolipram (IC50 540 +/- 67 nM, n = 3). 3. A close correlation (r = 0.95) was observed between suppression of IL-2 release by PDE inhibitors and inhibition of PDE4. In contrast, little correlation (r = 0.39) was observed between suppression of IL-2 release and their affinities for the high-affinity rolipram binding site (HARBS). 4. RP 73401 only inhibited partially (30-40%) Staph. A-induced incorporation of [3H]-thymidine into splenocyte DNA. The PDE3 inhibitor, siguazodan (10 microM), had little or no effect on IL-2 release or DNA synthesis. This concentration of siguazodan did not enhance the inhibitory action of RP 73401 on IL-2 release but potentiated its effect on DNA synthesis, increasing potency and efficacy. 5. Staph. A-induced DNA synthesis was only partially inhibited by anti-IL-2 neutralizing antibody, whereas dexamethazone (100 nM) and cyclosporine A (100 nM) completely blocked the response. 6. RP 73401 (IC50 6.3 +/ 1.9 nM, n = 4) was 140 fold more potent than rolipram (IC50 900 +/- 300 nM, n = 3) in inhibiting Staph. A-induced [3H]-thymidine incorporation into splenocyte DNA. 7. The results implicate a low-affinity form of PDE4 in the suppression of Staph. A-induced IL-2 release from murine splenocytes by PDE inhibitors. The data also indicate that mitogenic factors other than IL-2, whose elaboration or responses to which are regulated by PDE3 as well as PDE4, contribute to the superantigen-induced DNA synthesis. PMID- 9208144 TI - Modulatory role of 1,25 dihydroxyvitamin D3 on pancreatic islet insulin release via the cyclic AMP pathway in the rat. AB - 1. Previous studies have shown that vitamin D3 deficiency impairs the insulin response to glucose via an alteration of signal transduction pathways, such as Ca2+ handling and the phosphoinositide pathway. In the present study the adenylyl cyclase pathway was examined in islets from 3 independent groups: normal rats, 4 weeks-vitamin D3 deficient rats and one week-1,25 dihydroxyvitamin D3 (1,25(OH)2D3) treated rats. 2. We found that the very low rate of insulin release observed in vitamin D3 deficient rats could be restored in vitamin D3 deficient islets only with high concentrations of dioctanoyl-cyclic AMP (DO-cyclic AMP), whereas 1,25(OH)2D3 improved the sensitivity of the islets to this exogenous cyclic AMP analogue. 3. The beneficial effect of 1,25(OH)2D3 observed with or without DO-cyclic AMP was protein kinase A-dependent, since the addition of N-[2 (p-bromocinnamylamino) ethyl]-5-isoquinolinesulphonamide (H-89), a specific inhibitor of cyclic AMP-dependent protein kinases, decreased the insulin release of treated rats back to the level seen in vitamin D3 deficient islets. 4. The low rate of insulin release could not be consistently related to an alteration in cyclic AMP content of the islets. Indeed, low insulin response to a barium+theophylline stimulus observed in vitamin D3 deficient islets was paradoxically associated with a supranormal cyclic AMP content in the islets. 5. This paradoxical increase in cyclic AMP observed in these conditions could not be attributed to a lower total phosphodiesterase (PDE) activity, although the portion of Ca(2+)-calmodulin-independent PDE was predominant in islets from vitamin D3 deficient rats. 6. On the other hand, the higher cyclic AMP content of vitamin D3 deficient islets could be related to an increase in glucagon-induced cyclic AMP synthesis in relation to the hyperglucagonaemia previously observed in vitamin D3 deficient rats. Since higher concentrations of exogenous glucagon and higher endogenous cyclic AMP concentrations were required in vitro to restore insulin release to normal values, the cyclic AMP-dependent pathways that usually potentiate insulin secretion appeared to be less efficient in relation to an alteration in the post cyclic AMP effector system. 7. 1,25(OH)2D3 exerted a stimulating effect on insulin release via protein kinase A activation but reduced the supranormal cyclic AMP synthesis, thus exerting a differential modulatory influence on biochemical disturbances in islets induced by vitamin D3 deficiency. PMID- 9208145 TI - Pharmacological evaluation of IQM-95,333, a highly selective CCKA receptor antagonist with anxiolytic-like activity in animal models. AB - 1. The pyridopyrimidine derivative IQM-95,333 ((4aS,5R)-2-benzyl-5-[N alpha-tert butoxicarbonyl)L-tryptophyl] amino-1,3dioxoperhydropyrido[1,2-c]pyrimidine), a new non-peptide antagonist of cholecystokinin type A (CCKA) receptors, has been evaluated in vitro and in vivo in comparison with typical CCKA and CCKB receptor antagonists, such as devazepide, lorglumide, L-365,260 and PD-135,158. 2. IQM 95,333 displaced [3H]-CCK-8S binding to CCKA receptors from rat pancreas with a high potency in the nanomolar range. Conversely, the affinity of this new compound at brain CCKB receptors was negligible (IC50 > 10 microM). IQM-95,333 was a more selective CCKA receptor ligand than devazepide and other CCKA receptor antagonists. 3. Like devazepide, IQM-95,333 was a more potent antagonist of CCK 8S- than of CCK-4-induced contraction of the longitudinal muscle from guinea-pig ileum, suggesting selective antagonism at CCKA receptors. 4. IQM-95,333 and devazepide were also potent inhibitors of CCK-8S-stimulated amylase release from isolated pancreatic acini, a CCKA receptor-mediated effect. The drug concentrations required (IC50s around 20 nM) were higher than in binding studies to pancreas homogenates. 5. Low doses (50-100 micrograms kg-1, i.p.) of IQM 95,333 and devazepide, without any intrinsic effect on food intake or locomotion, blocked the hypophagia and the hypolocomotion induced by systemic administration of CCK-8S, two effects associated with stimulation of peripheral CCKA receptors. 6. IQM-95,333 showed an anxiolytic-like profile in the light/dark exploration test in mice over a wide dose range (10-5,000 micrograms kg-1). Typical CCKA and CCKB antagonists, devazepide and L-365,260 respectively, were only effective within a more limited dose range. 7. In a classical conflict paradigm for the study of anxiolytic drugs, the punished-drinking test, IQM-95,333, devazepide and L-365,260 were effective within a narrow dose range. The dose-response curve for the three drugs was biphasic, suggesting that other mechanisms are operative at higher doses. 8. In conclusion, IQM-95,333 is a potent and selective CCKA receptor antagonist both in vitro and in vivo with an anxiolytic-like activity in two different animal models, which can only be attributed to blockade of this CCK receptor subtype. PMID- 9208146 TI - Effect of papaverine on synaptic transmission in the guinea-pig ileum. AB - 1. The effect of papaverine, a well known smooth muscle relaxant, was investigated on neural transmission within the enteric nervous system. Segments of guinea-pig ileum were placed in a partitioned bath to enable drugs, including papaverine, to be applied to enteric nerve pathways without interfering with the recording of the smooth muscle contraction. Ascending excitatory enteric nerve pathways were activated by electrical field stimulation in the anal compartment (10 Hz for 2 s, 45 mA, 0.5 ms pulse duration) and the resulting contraction of the intestinal circular muscle in the oral compartment was recorded isotonically. 2. Tetrodotoxin (0.6 microM) and hexamethonium (100 microM) both abolished, or greatly reduced, the contractions when applied to either compartment indicating that nicotinic synapses are involved in this pathway. 3. Papaverine (0.3-30 microM) applied independently to each compartment depressed in a concentration dependent manner, the nerve-mediated contractions. The IC50 of this inhibitory effect was 3.53 microM for the oral and 4.76 microM for the anal compartments, respectively. Two other phosphodiesterase (PDE) inhibitors, 3-isobutyl-1 methylxanthine (IBMX 10-300 microM) and theophylline (30-1000 microM) added to the anal compartment also inhibited the nerve mediated contractions. Papaverine applied to the anal bath, after IBMX 100 microM (or theophylline 300 microM) further inhibited the nerve-mediated contractions, but was less effective than when applied alone. 4. Phentolamine (1 microM), an alpha-adrenoceptor antagonist, reduced the inhibitory effect of papaverine, but not that of IBMX (100 microM) or theophylline (300 microM). A combination of phentolamine and IBMX (or theophylline) prevented the inhibitory effect of papaverine. 5. Tetrodotoxin, but not papaverine or hexamethonium, inhibited the contraction elicited by electrical stimulation just anal to the partition indicating that papaverine did not affect the generation or conduction of nerve action potentials. 6. Verapamil (1 microM) and nifedipine (1 microM), two smooth muscle relaxants which act by blocking L type calcium channels, only inhibited the contractions when applied directly to the recording (oral) compartment. This indicates that L-type Ca2+ channels are probably not involved in synaptic transmission in these ascending pathways and thus that the PDE inhibitors do not inhibit synaptic transmission by acting on these channels. omega-Conotoxin GVIA (10 nM), a potent inhibitor of the N-type Ca2+ channels, blocked the nerve-mediated contractions applied to either compartment. Whether the PDE inhibitors exert their inhibitory actions via these channels remains to be established. 7. The results indicate that the PDE inhibitors, papaverine, IBMX and theophylline inhibit excitatory enteric neural pathways by depressing synaptic transmission. The inhibitory effect of papaverine (but not IMBX or theophylline) involves, at least in part, the release of noradrenaline from sympathetic nerves acting on alpha-adrenoceptors on enteric neurones. PMID- 9208147 TI - Differential activation of the epithelial and smooth muscle NK1 receptors by synthetic tachykinin agonists in guinea-pig trachea. AB - 1. The presence of tachykinin NK1 receptors have been shown in the epithelium and smooth muscle of guinea-pig airways. Previous data showed that substance P (SP), and the NK1 receptor agonist, [Sar9, Met (O2)11]-SP, relax guinea-pig tracheal tube preparations by stimulation of epithelial NK1 receptors and via nitric oxide (NO) release. However, the selective tachykinin NK1 receptor agonist, septide, was unable to produce this effect. The aim of the present study was to investigate the ability of a series of SP analogues to stimulate NK1 receptors of guinea-pig airway epithelium. 2. Isometric tension was recorded in isolated tracheal tube preparations in which compounds were administered intraluminally in the presence of phosphoramidon, indomethacin (both 1 microM) and the tachykinin NK2 receptor antagonist, SR 48,968 ((S)-N-methyl N-(4-acetyl-amino-4 phenylpiperidino)-2-(3,4-dichlorophenyl)butyl)benzam ide) (0.1 microM). Cumulative concentration-response curves were obtained in preparations under resting tone or in preparations precontracted with acetylcholine (ACh, 10 microM). 3. Contractile responses to low concentrations (0.1-10 nM) of substance P (SP) and the selective agonist of NK1 receptors, [Pro9]-SP. in non precontracted tracheae were higher in preparations pretreated with the NO synthase inhibitor, NG-monomethyl L-arginine (L-NMMA, 100 microM) than in preparations pretreated with its inactive enantiomer D-NMMA (100 microM). Tracheal tube preparations precontracted with ACh and pretreated with D-NMMA were relaxed by low concentrations of SP and [Pro9]-SP (0.1-10 nM). In contrast, after pretreatment with L-NMMA, SP and [Pro9]-SP contracted tracheae at all the concentrations tested. 4. Concentration-response curves to the NK1 receptor agonists, SP methyl ester, [Apa9-10]-SP and [pGlu6] SP (6-11) obtained in non precontracted tracheae were similar in the presence of either D-NMMA or L-NMMA. SP methyl ester, [Apa9-10]-SP and [pGlu6] SP (6-11) did not produce any relaxation, but instead, cause contractions in tracheal tube preparations precontracted with ACh and pretreated with D-NMMA. Concentration-response curves produced by all these agonists were similar in preparations precontracted with ACh and pretreated with L-NMMA or D-NMMA. 5. In guinea-pig tracheal tube preparations two groups of NK1 receptor agonists can be distinguished: one group, including [Pro9]-SP, stimulator epithelial NK1 receptors, the other group, including SP methyl ester, [Apa9-10]-SP and [pGlu6] SP (6-11), does not. One possible explanation for these findings and for the existence of compounds with a peculiar 'septide-like' pharmacological profile in the guinea-pig trachea could be the recently proposed phenomenon referred to as 'agonist-directed receptor trafficking'. PMID- 9208149 TI - Suppression by cyclosporin A of interleukin 1 beta-induced expression of group II phospholipase A2 in rat renal mesangial cells. AB - 1. We investigated whether cyclosporin A, a potent immunosuppressive drug, affects group II phospholipase A2. (PLA2; EC 3.1.1.4) induction in rat renal mesangial cells. 2. Previously we showed that the expression of group II PLA2 in rat renal mesangial cells is triggered by exposure of the cells to inflammatory cytokines such as interleukin 1 beta (IL-1 beta) or tumour necrosis factor alpha and agents that elevate cellular levels of cyclic AMP. Treatment of mesangial cells with IL-1 beta for 24 h induced PLA2 activity secreted into cell culture supernatants by about 16 fold. Incubation of mesangial cells with cyclosporin A inhibited IL-1 beta-induced PLA2 section in a dose-dependent fashion, with an IC50 value of 4.3 microM. Cyclosporin A did not directly inhibit enzymatic activity of PLA2. 3. Immunoprecipitation of radioactively labelled PLA2 protein from mesangial cell supernatants revealed that the inhibition of PLA2 activity is due to a suppression of PLA2 protein levels. This effect was preceded by a reduction of PLA2 mRNA steady state levels, as demonstrated by Northern blot analyses of total cellular RNA isolated from stimulated mesangial cells. 4. In order to evaluate whether cyclosporin A would affect the transcriptional activity of the PLA2 gene, we performed nuclear run on transcription experiments and provided evidence that the transcription rate of the PLA2 gene is reduced by cyclosporin A. 5. Previously we found that the nuclear transcription factor kappa B (NF kappa B) is an essential component of the IL-1 beta-dependent upregulation of PLA2 gene transcription. By electrophoretic mobility shift analysis, we demonstrated that cyclosporin A diminishes the formation of NF kappa B DNA binding complexes, thus suggesting that this transcription factor is a target for cyclosporin A-mediated repression of PLA2 gene transcription. 6. The data presented in this study strongly suggest that the cellular mechanism involved in the IL1 beta-dependent transcriptional upregulation of the PLA2 gene in mesangial cells is a target for the action of cyclosporin A. PMID- 9208148 TI - Pharmacological characterization of endothelin-induced rat pulmonary arterial dilatation. AB - 1. The aim of study was to characterize endothelin (ET)-induced vasodilation in isolated extrapulmonary rat arteries (EPA) and in intrapulmonary arteries (IPA) preconstricted with 1 microM phenylephrine. 2. The ET-3 (1 nM-100 nM)- and ET-1 (10 nM-100 nM)-induced transient vasodilatations in EPA were more potent than those in IPA. The vasodilatation induced by ET-3 (100 nM) was larger than that induced by ET-1 (100 nM). 3. Both the ETB antagonist, BQ788 (3 microM) and or endothelium denudation, but not the ETA antagonist, BQ123 (3 microM), abolished the vasodilation induced by ET-1 or ET-3 (100 nM each) in EPA and in IPA. The ATP sensitive K + channel blocker, glibenclamide (20 microM) and the nitric oxide synthase inhibitor, NG-monomethyl-L-arginine (L-NMMA, 1 mM) suppressed the ET induced vasodilatation in EPA and in IPA. 4. We conclude that the vasodilatation induced by endothelins is markedly reduced in rat isolated IPA, and suggest that the endothelial ETB-mediated vasodilatation varies depending on rat pulmonary arterial regions. Furthermore, ETB-mediated vasodilatation involves activation of ATP-sensitive K+ channels and of nitric oxide synthase in rat isolated EPA and IPA. PMID- 9208150 TI - Role of Ca(2+)-activated K+ channel in epithelium-dependent relaxation of human bronchial smooth muscle. AB - 1. To elucidate whether K+ channels play a role in the action of epithelium dependent bronchodilatation, we studied responses in human bronchial strips in the presence of indomethacin and NG-nitro-L-arginine methylester under isometric conditions, in vitro. 2. Mechanical removal of the epithelium increased the contractile responses to acetylcholine; the pD2 values increased from 5.0 +/- 0.2 to 5.9 +/- 0.3 (P < 0.001). This potentiation was abolished by iberiotoxin but not by apamin or glibenclamide. 3. In cascade bioassay, application of the bathing medium from dispersed, bronchial epithelial cells to epithelium-denuded bronchial strips decreased acetylcholine-induced contraction by 44 +/- 6%. This effect was reduced to 10 +/- 3% (P < 0.01) when the epithelial cells were pretreated with iberiotoxin, and to 4 +/- 1% (P < 0.001) when the epithelial cells were incubated with Ca(2+)-free medium containing [1,2-bis(2) aminophenoxy] ethane N,N,N',N'-tetraacetic acid-acetomethoxy ester. 4. In contrast, the bronchodilator effect of the medium bathing epithelial cells was not altered by the direct addition of iberiotoxin to epithelium-denuded tissues. 5. These results suggest that the Ca(2+)-activated K+ channel may play a role in the synthesis and/or release of smooth muscle relaxing factor, which is neither nitric oxide nor a cyclo-oxygenase product, from airway epithelial cells. PMID- 9208151 TI - Interactions between imidazoline compounds and sulphonylureas in the regulation of insulin secretion. AB - 1. Imidazoline alpha 2-antagonist drugs such as efaroxan have been shown to increase the insulin secretory response to sulphonylureas from rat pancreatic B cells. We have investigated whether this reflects binding to an islet imidazoline receptor or whether alpha 2-adrenoceptor antagonism is involved. 2. Administration of (+/-)-efaroxan or glibenclamide to Wistar rats was associated with a transient increase in plasma insulin. When both drugs were administered together, the resultant increase in insulin levels was much greater than that obtained with either drug alone. 3. Use of the resolved enantiomers of efaroxan revealed that the ability of the compound to enhance the insulin secretory response to glibenclamide resided only in the alpha 2-selective-(+)-enantiomer; the imidazoline receptor-selective-(-)-enantiomer was ineffective. 4. In vitro, (+)-efaroxan increased the insulin secretory response to glibenclamide in rat freshly isolated and cultured islets of Langerhans, whereas (-)-efaroxan was inactive. By contrast, (+)-efaroxan did not potentiate glucose-induced insulin secretion but (-)-efaroxan induced a marked increase in insulin secretion from islets incubated in the presence of 6 mM glucose. 5. Incubation of rat islets under conditions designed to minimize the extent of alpha 2-adrenoceptor signalling (by receptor blockade with phenoxybenzamine; receptor down-regulation or treatment with pertussis toxin) abolished the capacity of (+)- and (+/-) efaroxan to enhance the insulin secretory response to glibenclamide. However, these manoeuvres did not alter the ability of (+/-)-efaroxan to potentiate glucose-induced insulin secretion. 6. The results indicate that the enantiomers of efaroxan exert differential effects on insulin secretion which may result from binding to effector sites having opposite stereoselectivity. Binding of (-) efaroxan (presumably to imidazoline receptors) results in potentiation of glucose induced insulin secretion, whereas interaction of (+)-efaroxan with a second site leads to selective enhancement of sulphonylurea-induced insulin release. PMID- 9208152 TI - Inhibition of Ca2+ channel current by mu- and kappa-opioid receptors coexpressed in Xenopus oocytes: desensitization dependence on Ca2+ channel alpha 1 subunits. AB - 1. Desensitization of mu- and kappa-opioid receptor-mediated inhibition of voltage-dependent Ca2+ channels was studied in a Xenopus oocyte translation system. 2. In the oocytes coexpressing kappa-opioid receptors with N- or Q-type Ca2+ channel alpha 1 and beta subunits, the kappa-agonist, U50488H, inhibited both neuronal Ca2+ channel current responses in a pertussis toxin-sensitive manner and the inhibition was reduced by prolonged agonist exposure. 3. More than 10 min was required to halve the inhibition of Q-type channels by the kappa agonist. However, the half-life for the inhibition of N-type channels was only 6 +/- 1 min. In addition, in the oocytes coexpressing mu-opioid receptors with N type or Q-type channels, the uncoupling rate of the mu-receptor-mediated inhibition of N-channels was also faster than that of Q-type channels. 4. In the oocytes coexpressing both mu- and kappa-receptors with N-type channels, stimulation of either receptor resulted in a cross-desensitization of the subsequent response to the other agonist. Treatment of oocytes with either H-8 (100 microM), staurosporine (400 nM), okadaic acid (200 nM), phorbol myristate acetate (5 nM) or forskolin (50 microM) plus phosphodiesterase inhibitor did not affect either the desensitization or the agonist-evoked inhibition of Ca2+ channels. 5. These results suggest that the rate of rapid desensitization is dependent on the alpha 1 subtype of the neuronal Ca2+ channel, and that a common phosphorylation-independent mechanism underlies the heterologous desensitization between opioid receptor subtypes. PMID- 9208153 TI - Hydrogen peroxide-induced impairment of reactivity in rat isolated aorta: potentiation by 3-amino-1,2,4-triazole. AB - 1. In this study the impairment induced by hydrogen peroxide of vascular reactivity and the role of endogenous catalase in protection against this impairment was assessed in isolated rings of rat aorta. 2. Incubation with hydrogen peroxide at 1 mM, but not at 0.1 mM, for 15, 30 or 60 min followed by washout depressed, in a time-dependent manner, the subsequent ability of endothelium-containing and endothelium-denuded rings to contract to phenylephrine. 3. Incubation with 3-amino-1,2,4-triazole (50 mM, 90 min, followed by washout) to inhibit endogenous catalase had no effect by itself on subsequent phenylephrine-induced contraction. However, pretreatment with 3-amino-1,2,4 triazole did lead to a profound enhancement of the ability of hydrogen peroxide (1 mM, present for the final 30 min of the 90 min incubation, followed by washout) to depress phenylephrine-induced contraction in both endothelium containing and endothelium-denuded rings. 4. Incubation with hydrogen peroxide at 1 mM, but not at 0.1 mM, for 15, 30 or 60 min followed by washout inhibited, in a time-dependent manner, the subsequent ability of acetylcholine (10 nM-3 microM) to induce endothelium-dependent relaxation. Furthermore, incubation with hydrogen peroxide 1 mM (30 min, followed by washout) also inhibited relaxation induced by glyceryl trinitrate (1-100 nM) or isoprenaline (10 nM-3 microM) in endothelium denuded rings. 5. Incubation with 3-amino-1,2,4-triazole (50 mM, 90 min, followed by washout) had no effect by itself on relaxation induced by acetylcholine, glyceryl trinitrate or isoprenaline. In contrast, pretreatment with 3-amino-1,2,4 triazole led to profound enhancement of the ability of hydrogen peroxide (1 mM, present for final 30 min of the 90 min incubation) to block relaxation to acetylcholine, glyceryl trinitrate or isoprenaline. 6. On the basis of the actions of 3-amino-1,2,4-triazole, it is likely that endogenous catalase plays an important role in the protection of vascular reactivity of rat aorta against oxidant damage by high (1 mM) but not lower (0.1 mM) concentrations of hydrogen peroxide. The data are consistent with the promotion of non-selective damage to the vascular smooth muscle cells by hydrogen peroxide, but endothelial damage may also be sustained. PMID- 9208154 TI - Involvement of nitric oxide synthesis in hepatic perturbations induced in rats by a necrogenic dose of thioacetamide. AB - 1. The biological actions of nitric oxide (NO), a highly diffusible and short lived radical, range from signal transduction to cytotoxicity. The present study investigated whether NO is released in the course of liver necrosis and regeneration induced by a single necrogenic dose of thioacetamide (6.6 mmol kg-1 body wt) to rats. Samples of liver were obtained at 0, 3, 12, 24, 48, 72 and 96 h after thioacetamide administration. 2. Inducible nitric oxide synthase (iNOS) activity was determined in purified liver homogenates and a sharp 6 fold increase (P < 0.001) in iNOS activity was recorded at 48 h of intoxication, followed by a slight but progressive increase at 72 and 96 h. Changes in the expression of iNOS, as detected by its mRNA levels, were parallel to the NOS enzyme activity. Hepatocyte NO synthesis showed a progressive increase at 24, 48 and 72 h, to 8 (P < 0.001), 13 (P < 0.001) and 13 (P < 0.001) times the initial values, respectively. 3. In isolated Kupffer cells, where initial NO release was ten fold higher than in hepatocytes, a progressive increase was detected from 48 h which reached two fold of initial at 72 h of intoxication (192%; P < 0.001). Hepatic cyclic GMP concentration did not change significantly. However, mitochondrial aconitase activity decreased markedly at 12 and 24 h of intoxication showing a sharp increase towards normal values at 48 h which was maintained at 72 and 96 h. 4. As protein kinase C (PKC) is one of the likely candidates to mediate iNOS expression, translocation (activation) of PKC was assayed in hepatocytes, and a significant two fold increase (P < 0.001) between 48 and 96 h after thioacetamide intoxication was observed. When peritoneal macrophages from control rats were incubated with serum from thioacetamide-treated rats, a sharp increase in NO release was detected with serum obtained at 48 h, reaching at 96 h a value four fold (P < 0.001) that of the control. 5. These results suggest that iNOS activity and NO release play a role in the pathophysiological mechanisms that trigger post necrotic hepatocellular regeneration following thioacetamide administration. PMID- 9208155 TI - A study of the neurotoxic effect of MDMA ('ecstasy') on 5-HT neurones in the brains of mothers and neonates following administration of the drug during pregnancy. AB - 1. It is well established that 3,4-methylenedioxymethamphetamine (MDMA or 'ecstasy') is neurotoxic and produces long term degeneration of cerebral 5 hydroxytryptamine (5-HT) nerve terminals in many species. Since MDMA is used extensively as a recreational drug by young people, it is being ingested by many women of child bearing age. We have therefore examined the effect of administering high doses of MDMA to rats during pregnancy on the cerebral content of both the dams and the neonates. 2. MDMA (20 mg kg-1, s.c.) was injected twice daily on days 14-17 of the gestation period. The initial dose produced a marked hyperthermic response in the dam which was progressively attenuated in both peak height and area under the curve following further doses of the drug. The body weight of the dams decreased during the period of treatment. 3. There was a modest decrease in litter size (-20%) of the MDMA-treated dams. 4. The concentration of 5-HT and its metabolite 5-HIAA was decreased by over 65% in the hippocampus and striatum and 40% in the cortex of the dams 1 week after parturition. In contrast, the content of 5-HT and 5-HIAA in the dorsal telencephalon of the pups of the MDMA-treated dams was the same as that seen in tissue from pups born to control animals. 5. Administration of MDMA (40 mg kg-1, s.c.) to adult rats increased thiobarbituric acid reacting substances (TBARS) in cortical tissue 3 h and 6 h later, indicating increased lipid peroxidation. No increase in TBARS was seen in the cortical tissue of 7-10 day neonates injected with this dose of MDMA 3 h or 6 h earlier. 6. The data suggest that exposure to MDMA in utero during the maturation phase does not produce damage to 5-HT nerve terminals in the foetal rat brain, in contrast to the damage seen in the brains of the mothers. This may be due to MDMA being metabolized to free radical producing entities in the adult brain but not in the immature brain or, alternatively, to more effective or more active free radical scavenging mechanisms being present in the immature brain. PMID- 9208157 TI - Active and passive smoking: hazards for children. AB - This manuscript describes the tobacco industry's efforts to recruit active smokers among the adolescent population, the effects of environmental tobacco smoke on nonsmokers, and lists some steps pediatricians can take to influence smoking behavior. Six health effects result from passive smoking. Children exposed to environmental tobacco smoke have increased lower respiratory illness rates, especially in the first year of life, passive smoking is associated with increased rates of chronic middle ear effusion in children. Exposure to cigarette smoke is associated with small changes in pulmonary function. Exposure to environmental tobacco smoke increases an asthmatic child's exacerbations. Exposure to environmental tobacco smoke is a risk factor for sudden infant death syndrome. Passive exposure during childhood to a parents smoking increases a child's risk of leukemia and lymphoma during adulthood. Pediatricians and others who care for children must try to limit as much as possible, the exposure of children to the cigarette smoke produced by others. PMID- 9208156 TI - Antinociceptive and immunosuppressive effects of opiate drugs: a structure related activity study. AB - 1. Although it is well known that morphine induces significant immunosuppression, the potential immunosuppressive activity of morphine derived drugs commonly used in the treatment of pain (codeine, hydromorphone, oxycodone) has never been evaluated. 2. We evaluated in the mouse the effect of the natural opiates (morphine and codeine) and synthetic derivatives (hydromorphone, oxycodone, nalorphine, naloxone and naltrexone) on antinociceptive thresholds and immune parameters (splenocyte proliferation, Natural Killer (NK) cell activity and interleukin-2 (IL-2) production). 3. Morphine displayed a potent immunosuppressive effect that was not dose-related to the antinociceptive effect, codeine possessed a weak antinociceptive effect and limited immunosuppressive activity; nalorphine, a mu-antagonist and kappa-agonist, exerted a potent immunosuppressive effect, but had very weak antinociceptive activity. The pure kappa-antagonist nor-BNI antagonized the antinociceptive, but not the immunosuppressive effect of nalorphine. 4. Hydromorphone and oxycodone, potent antinociceptive drugs, were devoid of immunosuppressive effects. 5. The pure antagonists naloxone and naltrexone potentiated immune responses. 6. Our data indicate that the C6 carbonyl substitution, together with the presence of a C7-8 single bond potentiates the antinociceptive effect, but abolishes immunosuppression (hydromorphone and oxycodone). 7. The single substitution of an allyl on the piperidinic ring resulted in a molecule that antagonized the antinociceptive effect but maintained the immunosuppressive effect. 8. Molecules that carry modifications of C6, the C7-8 bond and C14, together with an allyl or caboxymethyl group on the piperidinic ring antagonized both the antinociceptive and the immunosuppressive effect of opiates and were themselves immunostimulants. PMID- 9208158 TI - Environmental epidemiology at the Medical Birth Registry of Norway; strengths and limitations. AB - A national registry of all pregnancy-outcomes was started in Norway. in 1967 to facilitate epidemiological surveillance. One aim was to detect increases in birth detect prevalences that was caused by new harmful exposures. Over the years several studies of unexpected time-trends and regional differences have been performed, but specific harmful exposures have so far not been detected. In a study of possible effects of the Chernobyl fallout, the registry enabled a detailed investigation of a possible increase in prevalence of birth detects that occurred specifically in the most exposed areas of Norway. The strength of this approach was the population coverage of the registry, while important limitations were incomplete reporting (outcome misclassification) and crude exposure information (exposure misclassification). While still taking advantage of the population coverage of the medical birth registry, a new generation of studies with more complete case ascertainment and individual exposure information are being prepared. One particularly interesting new option is to use molecular genetic tools to identity susceptible mothers or children. Collection and storage of biological specimens on a large scale is challenging, but in the search for gene-environment interactions, population-based studies are particularly attractive. PMID- 9208160 TI - Neurobehavioral aspects of lead neurotoxicity in children. AB - Neurobehavioural toxicity in occupational lead-exposure has typically not been observed at blood lead-concentrations (PbB) below 400 micrograms/l (e.g. 1, 2), whereas in environmentally exposed children such deficit has been reported to occur down to PbB of 100-150 micrograms/l and, perhaps, even below this range (4). Both cross-sectional and prospective studies have arrived at similar conclusions in this respect. The preferred endpoint in most such studies has been the IQ-measure, which has good psychometric qualities, is sufficiently well standardized to be comparable across studies, and exhibits attractive simplicity for the regulator in a public health context. Metaanalyses on both cross sectional and prospective studies in lead exposed children have concluded that a typical doubling of PbB from 100 to 200 micrograms/l is associated with an average loss of IQ of 1-3 points (3, 4). It should also be pointed out, however, that the IQ-focus has also interfered with systematic efforts to identify more specific lead-induced functional deficits by means of more detailed neurobehavioral analyses (5). Some neuropsychological findings in lead exposed children suggest that part of the impairment resembles performance deficit found to be characteristic for children presenting with signs and symptoms of attention deficit disorder. PMID- 9208159 TI - Blood lead levels in urban children of Katowice Voivodship, Poland: results of the population-based biomonitoring and surveillance program. AB - The paper presents the results of the large-scale blood lead levels survey in pre school urban children living in industrial area of Poland (Katowice Voivodship, Upper Silesian Industrial Zone-USIZ). The program, established in 1993, involves education, screening and medical care of case-children, as its major elements. Until December 1995 six thousand nine hundred sixty nine children aged 2-6 years have been examined in three towns (Chorzow, Kalowice, Sosnowiec). Geometric mean value of blood lead level (PbB) was slightly but not statistically significantly larger in boys (6.68 +/- 1.51 micrograms/dl) than in girls (6.58 +/- 1.54 micrograms/dl). In a multiple regression analysis the following variables explained variation in PbB: town (p = 0.0001), age (p = 0.005), floor on which apartment was located (p = 0.0001), number of siblings (p = 0.0001), apartment quality (p = 0.0001), carpet in a child's room (p = 0.0001), consumption of locally grown vegetables (p = 0.007), frequent trips outside the region (p = 0.0001). The results were verified with PbB as dichotomous variable. The occurrence of PbB above 10 micrograms/dl (frequency, 14.2%-17.2%) was associated with floor on which apartment was located, number of siblings, apartment's quality, the presence of carpet in child's room and frequent trips outside the region. The occurrence of PbB above 15 micrograms/dl (frequency: 2.5%-4.2% of children) was associated with the same variables and additionally, with the place of residence and intensity of vehicle traffic. The findings yield reliable population-based estimates of the risk of over-exposure of "non-hot-spot" urban children to environmental lead and highlight the important role of factors that could be classified as environmental and socio-economical determinants of blood lead level. Among environmental factors deposits of lead are still a problem in a densely populated industrial center of USIZ and the use of leaded gasoline adds to the magnitude of exposure. PMID- 9208161 TI - Methodological problems in assessing health-related, neuropsychological effects of lead absorption in a very low-level exposed area. AB - Methodological problems in the low level lead studies are reviewed using the Aarhus lead study as an example. It is shown that a lead effect can be found in an area where the background blood lead level is as low as 37 micrograms/l (geometric mean). Even in such an area it is worthwhile looking for populations at risk. It is shown that attrition causes confounding with a directional bias towards the null-hypothesis. Longitudinal studies are associated with this type of bias. Misclassification as for past exposures will also have a bias of this type. Studies depending on blood-lead measures are liable to have this type of bias. This is the case to a lesser degree in studies using cumulated indices. Subsamples of tooth dentin (circumpulpal dentin) satisfies the requirement for an index for cumulated lead absorption. Since lead absorption requires motor abilities, medical factors that are risk factors for motor development as well as for the other developmental outcome are also confined with a directional bias towards the null-hypothesis. It is suggested to exclude for such factors. Lead seems to be a ubiquitous noxious substance. Any community should have a strategy to minimize its effects. PMID- 9208162 TI - Monitoring of blood lead levels in Hungary. AB - Monitoring of blood lead levels (BLL) in children started in Hungary only in the 80's. A study on exposed areas of Budapest and Szolnok indicated a mean BLL higher than 20 micrograms/dl. Data from studies show a strong relationship between the BLLs of children and traffic. As a result of reduction of the lead content of gasoline, the BLLs of children decreased steeply in the early 90's. In 1994 a 6.9 micrograms/dl mean BLL was found in school children living in provincial towns, and 7.9% of the data exceeded the limit of 10 micrograms/dl. Recently a representative survey in preschool children (n = 1401, age 0-4 years) showed a mean BLL of 5.7 micrograms/dl, and only 4.7% exceeded the limit of 10 micrograms/dl. Despite this improvement nation-wide the condition is not satisfactory. Children are exposed to risk because of socioeconomic problems, carelessness, lack of public awareness. Such dangerous situations were created in the last two years by contamination of paprika powder, and battery dismantling without permission. PMID- 9208163 TI - Using the Hungarian Birth Defects Registry for surveillance, research and intervention. AB - The Hungarian Congenital Abnormality Registry (HCAR) was established in 1962 due to the growing public health importance of congenital abnormalities (CAs). The HCAR is a population-based registry; it collects and keeps permanent records of medical and personal information about malformed newborns and infants (notification of CAs Is mandatory for physicians) in order to develop baseline data for different types of CAs, to search for increases in the incidence of specific CAs, to provide rates, and identify geographic areas of concern for cluster investigation. In the last 25 years numerous studies have been carried out. In the 1970s two significant increases were noticed: i) in the rate of terminal transverse type of limb reduction defects and ii) in the rate of hypospadias. As a result of our epidemiological investigations and case-control studies we were able to identify the possible factors which caused these increases. In the first case, for example, the Ministry of Health changed the policy of using high oestrogens dosage (which increased the risk of this defect 6.1 times) for artificial abortions in 1978 and after this time the increase was stopped. In 1989-1990 a geographical cluster was found. An extremely high rate of cases with Down's syndrome (27%) was identified in a small village and the case control study and environmental investigations pointed suspicion on the excessive use of a chemical, trichlorfon at local fish farms. All mothers of babies with Down's syndrome had consumed contaminated fish during critical period. The use of this chemical was prohibited and no CAs were recorded after this time at this place. A special case-control surveillance system has been operating since 1980 in Hungary based on the HCAR. The objective of this system is to obtain information on pregnancy exposures and other risk factors for the study and identification of causes of CAs. The teratogenicity of several drugs taken during pregnancy has been examined so far, most of them ended up with negative results. Our system is suitable for surveillance, research and intervention. PMID- 9208164 TI - Association between ambient air concentrations of nitrogen dioxide and respiratory symptoms in children in Prague, Czech Republic. Preliminary results from the Czech part of the SAVIAH Study. Small Area Variation in Air Pollution and Health. AB - The primary objective of the SAVIAH, a multi-centre study funded by European Union, was to assess new methodology for study of small area health statistics and to implement it in epidemiological health statistics and geography. In Prague, the study has been conducted in two city districts with large variation in air pollution. Data at individual level (health symptoms and socio-economic circumstances of the family) were collected by questionnaires completed by parents of 3680 children aged 7-10 both resident and attending schools within the area (response rate 88%). Aggregated data for geographical areas were available from census and urban planning sources for 692 enumeration districts in the study area which were aggregated into 75 medium sized areas. Outdoor concentrations of nitrogen dioxide (NO2) were monitored by passive samplers. All these data were integrated into a geographic information system (GIS). Spatial distribution of air pollution was estimated by kriging and multiple regression modelling. These models explained about 80% of the variation in air pollution measured by passive samplers. GIS was then used to assign to individuals an exposure based on place of residence and school in order to conduct individual based analyses. Association between NO2 and life-time prevalence of wheezing and/or whistling, and wheezing/whistling in the last 12 months was studied by logistic regression. For both outcomes, school levels of NO2 were positively related to symptoms but home levels of NO2 showed a negative association. Logistic regression at individual level gives similar results as ecological analysis and multilevel modelling. Hierarchical model yielded somewhat wider confidence limits. Adjustment for parental behavioural and socio-economic factors did not affect these estimates substantially. This study demonstrated the power of the GIS methodology in studying the effects of complex environmental factors on respiratory health of children. PMID- 9208165 TI - A search for environmental and genetic background for neural tube defects: twenty five years of experience. AB - The present paper illustrates the author's 25-year experience in a step by step approach to the definition of environmental and genetic background of neural tube defects. Based on the birth defects registry, a complete ascertainment of all deliveries was performed in Southern Poland during two periods: 1970-1972, and 1979-1981. The birth prevalence of neural tube defects (NTD), as well as other CNS malformations was determined. The empiric recurrence risk was calculated as 3.2% +/- 1.6. Based on this figure, the relative risk (RR = 37.6 p < 0.001) and heritability (h2 = 74.7 +/- 6.7) were estimated. Our own modification of Morton's complex segregation analysis was applied. Three Mendelian (dominant, additive and recessive) and one multifactorial model were tested. The results did not provide a clear cut discrimination between different models; however the lowest x2 value was obtained for additive inheritance with 61% of penetrance and the frequency of sporadic cases equaled 55%. A search for genetic markers did not support the hypothesis that HLA-A,B,C loci are equivalents of T/t like locus in mice. The results of the study on transcobalamine levels in amniotic fluid may suggests that different transcobalamine metabolism reflects phenotypic expression of genetic susceptibility to NTD development. Current research status and future perspectives on genetic and environmental background of NTD are also presented. PMID- 9208166 TI - The analysis of registry data in relation to various different types of hypothesis regarding the geographical distribution of disease. AB - Disease registries will often contain the addresses of cases included in the registry. If the registry includes information on all cases, or deaths, occurring in a defined geographical area and time period and if there is a postcode/zip code or map reference for each case it is possible to carry out a variety of different types of geographical analysis that may give clues to the aetiology of the disease. For such analyses it will usually also be necessary to have population data for the region covered by the registry and for separate sub regions within it. In this paper we review types of analysis that may be applied to such data and give references to examples of applications and the statistical methods used. These include, first, methods of presenting incidence rates, and particularly the use of maps; of particular concern is the development of methods for presenting data that take into account the problems of rates calculated for small populations and which may therefore happen to be high or low simply by chance. Secondly, we consider, the analysis of "clustering" and "clusters" of cases of disease. These problems have been the subject of considerable methodological development in recent years. Analyses of clustering address the question of whether there is a general tendency for there to be aggregations of cases or areas of high incidence the analysis of clusters is concerned with problems of detecting specific locations where there are unusual aggregations of cases. The third type of problem considered here is whether there are, within the registry region, aetiological factors that vary geographically with consequent variations in disease incidence in different sub-regions. Where there is geographical variation it may be possible to use regression analysis to relate such variation to factors such as socio-economic status or levels of some environmental hazard. Finally we consider the problem of determining whether disease rates in certain areas may be related to distance from the source of some potential causative agent. PMID- 9208167 TI - Assessment of Polish population exposure to lead and cadmium with special emphasis to the Katowice Province on the basis of metal concentrations in environmental compartments. AB - Polish data of lead and cadmium concentrations in such environmental compartments as: air, dust, soil, diet and drinking water were presented in the paper. Special emphasis was placed to the Upper Silesian industrial Region, the central part of the Katowice Province, the most polluted area in Poland. An attempt was made to assess human exposure to heavy metals. According to the available data it can be concluded that the WHO tolerable intake (ADI, PTWI) could be exceeded in an extremely disadvantageous situation occurring in the Katowice Province. Furthermore, a critical review of heavy metal standards in soil, dustfall and edible plants, obligatory in Poland, was presented. The author states that there are still some inconsistencies in current regulations in Poland. In order to remove them, an interdisciplinary board consisting of such disciplines representatives as: medicine, toxicology, nutritional science, law and protection of soils used for agricultural purposes should be appointed. PMID- 9208168 TI - Cytochrome P450-catalyzed oxidation of halobenzene derivatives. PMID- 9208169 TI - Mechanism of dithiocarbamate inhibition of apoptosis: thiol oxidation by dithiocarbamate disulfides directly inhibits processing of the caspase-3 proenzyme. AB - Dithiocarbamates (DCs) have been reported to be potent inhibitors of apoptosis in several different model systems, which suggests a target common to the apoptotic machinery. Without further investigation, this has been assumed to reflect an antioxidant activity of the DCs. However, we have recently shown that DCs exert prooxidant effects on T cells [Nobel et al. (1995) J. Biol. Chem. 270, 26202 26208], which are dependent on their transfer of external copper into the cells and can be inhibited by the inclusion of high-affinity external copper chelators in the medium. Investigating antiapoptotic actions of DCs, we found that inclusion of a membrane-impermeable copper chelator severely compromised the inhibitory activity of reduced DCs. Since copper can promote DC oxidation to the respective DC disulfides, the inhibitory effect on lymphocyte apoptosis might be mediated by the DC disulfides. In agreement with this we observed that DC disulfides were more potent inhibitors of T cell apoptosis than their reduced counterparts. Inhibition of apoptosis by DC disulfides correlated with the inhibition of caspase-3 proenzyme processing and activation. Similar results were obtained in a cell-free model system of caspase-3 activation. Significantly, dithiothreitol reduction of the DC disulfide abolished its inhibition of in vitro proenzyme processing, thereby demonstrating thiol-disulfide exchange between the DC disulfide and a free thiol group on an activator(s) of caspase-3. Since T cell apoptosis involves the generation of mature caspase-3 and requires caspase-3-like activity, we propose that (1) DC disulfides are the active agents behind DC inhibition of apoptosis and (2) their site of action is the proteolytic activation of this enzyme. These findings also reveal the potential for other thiol-oxidizing toxicants to inhibit apoptosis by preventing the proteolytic activation of caspases. PMID- 9208170 TI - Simulation of the induction of oxidation of low-density lipoprotein by high copper concentrations: evidence for a nonconstant rate of initiation. AB - Kinetic simulation can help obtain deeper insight into the molecular mechanisms of complex processes, such as lipid peroxidation (LPO) in low-density lipoprotein (LDL). We have previously set up a single-compartment model of this process, initiating with radicals generated externally at a constant rate to show the interplay of radical scavenging and chain propagation. Here we focus on the initiating events, substituting constant rate of initiation (Ri) by redox cycling of Cu2+ and Cu+. Our simulation reveals that early events in copper-mediated LDL oxidation include (1) the reduction of Cu2+ by tocopherol (TocOH) which generates tocopheroxyl radical (TocO.), (2) the fate of TocO. which either is recycled or recombines with lipid peroxyl radical (LOO.), and (3) the reoxidation of Cu+ by lipid hydroperoxide which results in alkoxyl radical (LO.) formation. So TocO., LOO., and LO. can be regarded as primordial radicals, and the sum of their formation rates is the total rate of initiation, Ri. As experimental information of these initiating events cannot be obtained experimentally, the whole model was validated experimentally by comparison of LDL oxidation in the presence and absence of bathocuproine as predicted by simulation. Simulation predicts that Ri decreases by 2 orders of magnitude during lag time. This has important consequences for the estimation of oxidation resistance in copper-mediated LDL oxidation: after consumption of tocopherol, even small amounts of antioxidants may prolong the lag phase for a considerable time. PMID- 9208171 TI - Synthesis, characterization, and immunochemical detection of O6-(carboxymethyl) 2'-deoxyguanosine: a DNA adduct formed by nitrosated glycine derivatives. AB - O6-(Carboxymethyl)-2'-deoxyguanosine (O6-CMdG) is formed in DNA by nitrosated glycine derivatives and appears to be nonrepairable by O6-alkylguanine transferases. O6-CMdG has been synthesized by an unambiguous route involving the introduction of a methyl glycolate moiety at C6 of a 3',5'-bis-O (methoxyacetyl)dGuo derivative by displacement of a quinuclidinium ion. Methanolysis of the methoxyacetyl groups and calcium hydroxide-mediated hydrolysis of the methyl ester afforded the calcium salt of O6-CMdG in good yield. A similar route was used to synthesize O6-(carboxymethyl)guanosine (O6 CMGuo), which was used to prepare protein conjugates for immunization. Rabbit antisera were prepared, and a quantitative competitive ELISA was developed which showed 50% inhibition at 2 pmol of O6-CMdG/ well. O6-CMGuo was 30 times less cross-reactive (50% inhibition at 60 pmol/well), and normal nucleosides and carboxymethylated purines did not cross-react to any significant extent. The antiserum was also used to prepare reusable immunoaffinity columns which retained O6-CMdG. The binding of O6-CMdG was so strong that conditions used to elute the adduct (1 M trifluoroacetic acid) resulted in partial hydrolysis (becoming quantitative on heating) of the glycosidic bond to give O6-CMguanine which was detected by HPLC with fluorescence detection. DNA treated with azaserine (5 mmol), N-(N'-acetyl-L-prolyl)-N-nitrosoglycine (5 mmol), and potassium diazoacetate (5 mmol) afforded O6-CMdG at levels of 7.3, 393.9, and 496 mumol of O6-CMdG/mol of dG. The antiserum also recognized O6-CMdG in intact DNA. PMID- 9208172 TI - Accurate and sensitive quantitation of N7-methyldeoxyguanosine-3'-monophosphate by 32P-postlabeling and storage-phosphor imaging. AB - As N7-methyldeoxyguanosine-3'-monophosphate (N7-MedGp) is the major, persistent DNA lesion generated by methylating agents, a combined HPLC/32P-postlabeling assay has been developed to quantitate this adduct in human DNA. N7-MedGp was purified from normal nucleotides by anion-exchange chromatography followed by reverse-phase HPLC procedures. The adduct was then 32P-postlabeled and resolved by two-dimensional TLC for detection and quantitation by storage-phosphor imaging. The effect of conditions used for DNA purification and digestion on the recovery of N7-MedGp has been investigated. Extended, raised temperature incubations normally employed during DNA purification were demonstrated to result in considerable loss of adduct through depurination after 22 h at 65 and 37 degrees C (82% and 20% loss, respectively), but depurination was reduced to 5% if the incubation was performed at either 4 or 22 degrees C. Similarly, close to optical recovery (83%) of N7-MedGp was achieved after DNA digestion by incubating at 4 degrees C, pH 7.4, for 18 h in the presence of micrococcal nuclease and calf spleen phosphodiesterase from Sigma and Boehringer Mannheim, respectively. Overall, the recovery of N7-MedGp was 40%, resulting in a detection limit of 1.3 fmol which is equivalent to 0.16 mumol of adduct/mol of 2'-deoxyguanosine-3' monophosphate (dGp) when analyzing 10 micrograms of DNA. The N7-MedGp content of DNA that had been methylated in vitro using 0, 16, and 80 microM N-methyl-N nitrosourea (NMU) was determined by 32P-postlabeling to be 12, 112, and 671 mumol of N7-MedGp/mol of dGp. Electrochemical detection of N7-methylguanine (N7-MeG) after HPLC purification measured approximately 2-fold higher levels, i.e., 25, 225, and 1080 mumol of N7-MeG/mol of Gua, at each NMU concentration, respectively. The levels of N7-MedGp in the white blood cell (WBC) DNA of patients receiving a single dose of 5-(3,3-dimethyl-1-triazeno)imidazole-4 carboxamide (DTIC) chemotherapy were determined by 32P-postlabeling. Maximum levels were found 4-6 h after treatment, and in two out of four individuals adduct levels were decreased by 21 h. Prior to treatment, N7-MedGp was detectable in WBC DNA in two out of the four individuals indicating that nontherapeutic exposure to methylating agents had occurred. PMID- 9208173 TI - New strategy for the construction of single-stranded plasmids with single mutagenic lesions. AB - Single-stranded DNA vectors containing single adducts offer a unique opportunity to study the biochemistry and genetics of trans lesion synthesis, a process during which a DNA polymerase synthesizes across a lesion. We describe a new and general strategy to produce high-quality single-stranded plasmids containing a single adduct within a predetermined sequence context starting with a short oligonucleotide containing the lesion of interest. These vectors are isolated from the corresponding double-stranded constructs by selective enzymatic degradation in vitro of the nonadducted uracil-containing strand. Efficient and complete removal of this strand was achieved using uracil DNA glycosilase to generate AP sites followed by the action of the AP endonuclease associated with exonuclease III and the robust 3'-->5' exonuclease activity associated with T7 DNA polymerase. We show the utility of these constructs for the study of trans lesion synthesis in vitro and in vivo in the case of the highly carcinogenic N-2 acetylaminofluorene adducts located within frameshift mutation hot spots. The possibility to construct both single-stranded and double-stranded plasmids, with the same origin of replication (i.e., ColE1), will allow a direct comparison between single-stranded and double-stranded DNA replication in site-specific mutagenesis studies. PMID- 9208174 TI - Aflatoxin B1 8,9-epoxide hydrolysis in the presence of rat and human epoxide hydrolase. AB - Aflatoxin B1 (AFB1) must be activated to the electrophilic AFB1 exo-8,9-epoxide to be genotoxic and carcinogenic. A role for epoxide hydrolase in detoxication has been suggested but never directly addressed. In light of recent studies determining the instability of AFB1 exo-8,9-epoxide in H2O, a role for epoxide hydrolase appears dubious. Rat liver or recombinant rat epoxide hydrolase provided an enhancement to the already fast hydrolysis rate of up to 22%. Purified human epoxide hydrolase provided no detectable enhancement to the rate of chemical hydrolysis. Some reduction in the genotoxicity of AFB1 was observed when the ratio of rat epoxide hydrolase to cytochrome P450 was high (approximately 50-fold). An 80-fold excess of human epoxide hydrolase over cytochrome P450 only produced an effect of approximately 25% inhibition. It appears, therefore, that there is little evidence to support a role for epoxide hydrolase in the detoxication of AFB1. PMID- 9208175 TI - Conformational studies of stereoisomeric tetrols derived from syn- and anti dibenzo[a,l]pyrene diol epoxides. AB - An understanding of the conformational behavior of the stereoisomeric tetrols at the 11,12,13,14-positions of dibenzo[a,l]pyrene (DB[a,l]P) is essential for the spectroscopic identification of DNA adducts derived from the biologically highly active fjord region syn- and anti-DB[a,l]P-11,12-diol 13,14-epoxides. Conformational effects are expected to play an important role in DNA-DB[a,l]P diol epoxide reactivity, base-sequence specificity, and conformation dependent repair. The results of conformational studies on trans-anti-, cis-anti-, and cis syn-DB[a,l]P tetrol isomers are presented and compared to the results obtained previously for trans-syn-DB[a,l]P tetrol (Carcinogenesis 17, 829-837, 1996). Molecular mechanics, dynamical simulations, and semiempirical calculations of electronic transitions are used to interpret the low-temperature fluorescence spectra and 1H NMR data. Molecular dynamics simulations (in vacuo) identified two conformers (I and II) for each of the tetrol isomers; in all conformations the aromatic ring system is severely distorted. Fluorescence line-narrowing (FLN) spectroscopy identified two distinct conformational species for the trans-anti isomer, one occurring in ethanol and the other occurring in a glycerol/water matrix. The corresponding structures are assigned based on the S1<--S0 transition energies calculated for conformers I and II, respectively. 1H NMR spectroscopy confirmed the structure of conformer I at room temperature. In contrast to trans syn-DB[a,l]P tetrol (where the major conformation was identified as a boat structure), both conformations of trans-anti-DB[a,l]P tetrol feature a half-chair structure for the cyclohexenyl ring with different orientations of the hydroxyl groups. For cis-anti- and cis-syn-DB[a,l]P tetrols, only a single conformer is detected by FLN spectroscopy. The NMR results for the latter appear to be most consistent with a mixture of two half-chair conformers I and II, while for the cis-anti isomer a flattened, boatlike conformation was observed. The generally good agreement between the NMR coupling constants and those estimated theoretically indicates that these structures should serve as good starting points for spectroscopic or computational studies of DNA adducts derived from DB[a,l]P diol epoxides. PMID- 9208176 TI - Stereoselective activation of dibenzo[a,l]pyrene and its trans-11,12-dihydrodiol to fjord region 11,12-diol 13,14-epoxides in a human mammary carcinoma MCF-7 cell mediated V79 cell mutation assay. AB - Dibenzo[a,l]pyrene (DB[a,l]P) represents the most potent carcinogenic polycyclic aromatic hydrocarbon (PAH) yet discovered. Like other PAHs, DB[a,l]P requires metabolic activation to exert its mutagenic and/or carcinogenic activity. In the human mammary carcinoma cell line MCF-7, DB[a,l]P is stereoselectively metabolized to the (-)-anti- and (+)-syn-DB[a,l]P-11,12-diol 13,14-epoxides (DB[a,l]PDE) which both bind extensively to deoxyadenosine residues in DNA. To further characterize the underlying mechanism of its strong carcinogenicity, the relationship between DNA binding and mutagenicity of DB[a,l]P was determined. Racemic DB[a,l]P-11,12-dihydrodiol and the two individual (+)- and (-) enantiomers, the metabolic precursors of the stereoisomeric fjord region dihydrodiol epoxides, were also investigated. Induction of mutations at the HPRT locus was measured in a MCF-7 cell-mediated Chinese hamster V79 cell mutation assay. The parent hydrocarbon, (+/-)-DB[a,l]P-11,12-dihydrodiol, and (-)-DB[a,l]P 11,12-dihydrodiol were highly mutagenic under the assay conditions. In contrast, (+)-DB[a,l]P-(11S,12S)-dihydrodiol was not mutagenic using MCF-7 cells as the metabolic activating system. Analysis of DNA adducts in the same experiments revealed that MCF-7 cells treated with (-)-DB[a,l]P-11,12-dihydrodiol formed exclusively (-)-anti-DB[a,l]-PDE adducts whereas cells treated with (+)-DB[a,l]P 11,12-dihydrodiol did not contain detectable levels of DNA adducts. These results suggest that specific cytochrome P450 enzymes may have high stereoselectivity for activation of the two DB[a,l]P-11,12-dihydrodiol enantiomers, and this may play an important role in the metabolic activation of the strong carcinogen DB[a,l]P in human cells. PMID- 9208177 TI - Association of dehydromonocrotaline with rat red blood cells. AB - The association of radiolabeled monocrotaline pyrrole (DHM) with red blood cell (RBCs) ghosts, globins, and heme was examined to determine their role in the transport and stabilization of this hepatic produced putative toxic metabolite of the pyrrolizidine alkaloid monocrotaline (MCT). Rats were administered 5 mg of DHM/kg, i.v., and RBCs and plasma were harvested at 4 and 24 h. Extensive washing of the RBCs with isotonic phosphate buffer did not decrease the amount of radioactivity associated with the cells. The level of DHM equivalents recovered in the RBCs did not decrease between 4 and 24 h, while the plasma levels, which were 29- and 75-fold lower, respectively, decreased from 5.0 to 2.2 nmol of DHM equiv/g of plasma. Globin chains were found to contain 383 and 453 pmol of DHM equiv/mg of protein, respectively. Rats receiving 10 mg of DHM/kg, i.v., with RBCs collected at 2 h, had approximately double the level of radioactivity associated with their RBCs in addition to 2 times the amount of adducts on the globin chains. Globins and ghosts plus heme (2 h) contained 69% and 2% of the radioactivity, respectively. Globin chains treated with an acidic ethanol solution containing AgNO3 resulted in the removal of 31% of the associated radioactivity. GC/ MS and TLC separation of AgNO3-displaced material revealed the presence of the ethyl ether derivatives of 7-hydroxy-1-(hydroxymethyl)-6,7 dihydro-5H-pyrrolizine. The HPLC separation of globin chains revealed that the majority of radioactivity coeluted with the beta-chains. In conclusion, this study found that the administration of radiolabeled DHM resulted in extensive radioactive labeling of RBCs; similar findings have been reported for [14C]MCT. PMID- 9208178 TI - Modification of horse heart cytochrome c with trans-2-hexenal. AB - Horse heart cytochrome c reacting with trans-2-hexenal was used as a simple model of the nonspecific interactions of proteins with 2-alkenals. The reaction mixtures containing relatively high concentrations of the protein and aldehyde were characterized using visible spectrophotometry, fluorescence, and circular dichroism measurement, capillary isoelectric focusing, size-exclusion chromatography, polyacrylamide gel electrophoresis, and mass-spectrometric techniques. The mass-spectrometric data indicate that cytochrome c becomes modified with one or two molecules of hexenal as the major reaction product. The modified species with a correspondingly lowered isoelectric point were detected through capillary isoelectric focusing. The results of proteolytic studies indicate nonspecific modifications. Significant quantities of the oligomeric forms of hexenal-modified protein were also observed electrophoretically. PMID- 9208179 TI - Occupational sensitization to laboratory animals. PMID- 9208180 TI - Heterogeneity of airway hyperresponsiveness. PMID- 9208181 TI - Exposure of laboratory animal workers to airborne rat and mouse urinary allergens. AB - BACKGROUND: Laboratory animal workers are at high risk of developing occupational allergy. Little is known about the relationship between levels of exposure and the risk of developing laboratory animal allergy. Since laboratory animal work comprises a large number of different-often short lasting-tasks, it is of interest to assess which activities are associated with high, low or intermediate levels of allergen exposure. OBJECTIVE: To develop and evaluate highly sensitive immunoassays in order to quantify rat and mouse urinary allergens in airborne dust sampled during short-lasting tasks. METHODS: Personal air dust samples were taken during full-shift periods as well as during specific tasks in seven laboratory animal facilities. Two sandwich enzyme immunoassays were developed, using rabbit antisera against rat and mouse urinary proteins. The rabbit antibodies were analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting and compared with IgE antibodies from sensitized laboratory animal workers. RESULTS: The rabbit antibodies were highly specific for rat and mouse urinary proteins and reacted with all IgE binding allergens in either urinary protein preparation. The assays for rat and mouse urine were very sensitive, with detection limits of 0.075 ng/mL. The coefficient of variation of the analysis was 12.9% for both assays. Animal caretakers appeared to experience the highest exposure to aeroallergens. A large variation in exposure within jobs was found, due to differences between tasks performed during the sampling day and the facility worked at. The highest exposure levels were found during removal of contaminated bedding from the cages. However, rat and mouse allergen exposure levels during this task varied enormously between facilities, 1.1-158 ng eq/m3 and 0.63-2000 ng eq/m3, respectively. CONCLUSION: Both sandwich immunoassays are highly specific and sensitive and are able to identity tasks of relatively short duration with high, medium and low exposure to airborne rat and mouse urinary allergens. PMID- 9208182 TI - Seasonal differences in airway hyperresponsiveness in asthmatic patients: relationship with allergen exposure and sensitization to house dust mites. AB - BACKGROUND: The degree of airway hyperresponsiveness in allergic asthmatic patients may be influenced by changes in environmental exposure to inhalant allergens. OBJECTIVE: This study investigates the relationship between seasonal changes in exposure to house dust mite (HDM) allergens and non-specific airway hyperresponsiveness in asthmatic patients with multiple sensitizations to inhaled allergens. METHODS: In 43 asthmatic patients sensitized to several inhalant allergens, lung function (FEV1), airway hyperresponsiveness (PC20 histamine), serum total IgE, house dust mite (HDM) specific IgE and number of peripheral blood eosinophils were measured during autumn 1990 (September-November) and spring 1991 (March-May). During each season, floor dust samples were collected twice from living- and bedrooms and the concentration of the HDM allergens Der p 1 and Der p 2 determined. RESULTS: More severe airway hyperresponsiveness (lower PC20 histamine) during autumn was only found in patients sensitized to HDM (n = 32; autumn: 2.05 mg/mL, spring: 4.51 mg/mL (geometric means), P < 0.01), whereas in patients not sensitized to HDM (n = 11) similar values were observed in both seasons (3.44 and 4.52 mg/mL, respectively, P = 0.56). More severe airway hyperresponsiveness of HDM sensitized patients in autumn was significantly associated with higher Der p 1 concentrations in floor dust. Aside from airway hyperresponsiveness, seasonal changes in serum total IgE and number of peripheral blood eosinophils were seen in patients sensitized to HDM. CONCLUSIONS: In allergic asthmatic patients, airway hyperresponsiveness may increase during autumn, depending on sensitization to HDM and an increase of exposure to HDM allergen. PMID- 9208183 TI - Clinical characteristics of peanut allergy. AB - BACKGROUND: Current clinical advice regarding peanut allergy is based on small series of patients. OBJECTIVE: To determine, in a large group of peanut allergic subjects, the patterns of clinical severity, symptom progression and availability and use of rescue medications. METHODS: Questionnaire study of 622 self-reported allergic subjects. RESULTS: A total of 406 patients (66%) reported symptoms on contact with peanut. Only 121 (19%) had been knowingly exposed to peanut before the first documented reaction, implying a high frequency of occult sensitization. Severe symptoms were more common in adults. Abdominal symptoms were significantly associated with collapse. Fifty per cent reported reactions in the previous year. Only 82 (13%) had been admitted to hospital because of a reaction. Adrenaline was carried in some form by 65% though only 78 subjects (12.5%) had ever received injected adrenaline. Only 18/43 subjects (41%) who collapsed were given adrenaline. Skin-prick test weal size correlated weakly with severity but there were large overlaps between the groups. Peanut-specific IgE peaked in the teenage group, but did not correlate with severity. CONCLUSIONS: Peanut allergy is characterized by more severe symptoms than other food allergies and by high rates of symptoms on minimal contact. Skin-prick testing and peanut-specific IgE levels do not predict clinical severity. Avoidance of peanut is difficult. Many people suffering severe relations are inadequately treated. Sufferers need education and training in the use of rescue medication. PMID- 9208184 TI - Occupational allergy in greenhouse workers: sensitization to Tetranychus urticae. AB - BACKGROUND: Tetranychus urticae (TU) is a macroscopic mite which is found infesting a large number of plants of economic interest. It has rarely been described as a cause of occupational allergic disease in agricultural workers. OBJECTIVE: To describe TU sensitization in greenhouse workers attending the outpatient allergy unit and its clinical associations, and to characterize the allergens involved. MATERIALS AND METHODS: Twenty-four consecutive carnation greenhouse workers with allergy-related symptoms, referred to our outpatient clinic during a 6-month period, were included. We made the diagnostic extract from carnation leaves heavily infested with TU. Skin-prick test, specific IgE measurement and bronchial provocation test with TU extract were carried out in all subjects. Allergen characterization was achieved by SDS-PAGE (sodium dodecylsulfate-polyacrylamide gel electrophoresis) and immunoblotting. RESULTS: Sixteen patients (66%) presented positive skin-prick test and specific IgE and were diagnosed allergic to TU. Fifteen patients suffered from bronchial asthma, 14 rhinitis and five urticaria. Twelve exhibited positive bronchial provocation test to the TU extract. On RAST-inhibition studies, there was no evidence of crossreactivity between TU extract and D. pteronyssinus. An allergen at 19 kDa was determined in the TU extract by SDS-PAGE immunoblotting studies. CONCLUSION: TU could be an important occupational allergen in greenhouse workers showing allergic symptomatology. There is no crossreactivity between this mite and the house dust mite D. pteronyssinus. PMID- 9208185 TI - The most common phenotypes of sensitization to inhalant allergens in childhood. AB - BACKGROUND: Atopic individuals are frequently sensitized to a limited number of certain allergens, although most of them are exposed to multiple inhalant allergens in daily life. OBJECTIVE: We investigated the hypothesis that observed common patterns of sensitization might occur with similar frequency within two independent study populations of school-children. METHODS: The results were derived from skin-prick tests conducted on two large samples of children (study 1: n = 583; study 2: n = 1099) examined with the same panel of six inhalant allergens. RESULTS: In order to ensure that the comparison was uniform, the younger subpopulation of study 1 (n = 147) was compared with the sample of study 2 (n = 374). The highest frequency for monosensitization was found for sensitization to Dermatophagoides pteronyssinus (study 1: 18.4%, study 2: 20.3%), followed by monosensitization to grass pollens (study 1: 12.2%; study 2: 8.8%). Using multiple logistic regression for each specific sensitization, a significantly increased relative risk of sensitization to hazel pollens (study 1 OR 5.9; study 2 OR: 24.3) appeared to be associated with sensitization to birch pollens. The same applied to dog dander (study 1 OR: 7.3; study 2 OR: 2.6), which showed an association with sensitization to cat dander. CONCLUSION: In summary, our data suggest that certain clusters of monosensitization and polysensitization to common inhalant allergens exist among a given population. This may well be a reflection of diversity in disposition to specific sensitization and/or antigen crossreactivity. From a practical point of view the data also might help in counselling parents of allergic children. PMID- 9208186 TI - Mast cell derived heparin activates the contact system: a link to kinin generation in allergic reactions. AB - Contact activation occurs when plasma comes in contact with negatively charged manmade surfaces but no substance that initiates contact activation in vivo has been identified. We have isolated a mast cell heparin proteoglycan (MC-HepPG) from a Furth mouse mastocytoma-derived cell line that is analogous to human tissue-type mast cell HepPG. This material and other glycosaminoglycans (GAGs) were tested for their ability to accelerate the reciprocal activation of factor XII and prekallikrein and the autoactivation of factor XII. Quantitative analysis showed the MC-HepPG to be as active as dextran sulfate on a weight basis; hog intestine heparin, dermatan sulfate, keratan polysulfate and chondroitin sulfate C were less active, other sulfated polysaccharides were essentially inactive. Incubation of MC-HepPG in 1:4 diluted plasma resulted in complete cleavage of high molecular weight kininogen in a factor XII-dependent reaction. All of the MC HepPG dependent reactions described above were inhibited by preincubation of MC HepPG with heparinase I and II but not by pretreatment with heparitinase, chondroitinase ABC or the serine protease inhibitor aPMSF thus indicating that heparin proteoglycan is indeed acting as an initiating 'surface'. We analysed the proteoglycan preparation by HPLC gel filtration. Fractions spanning a molecular weight range of > 400000-8000 were active initiators. Comparison of the chromatograms obtained before and after cleavage of GAG side chains from the protein core suggested that dissociated GAGs in the MW range 69000-17000 are the most active species rather than the complete proteoglycan. MC-HepPG GAGs therefore represent a physiologic macromolecule with activity comparable to non physiological surfaces in a purified system and with the capability to induce activation of the contact system in diluted plasma. Its ability to promote kinin generation links cellular and humoral inflammatory responses in the perivasculature and provides a possible explanation for the elevated kinin levels observed after allergen exposure. PMID- 9208187 TI - Expression of adhesion molecules on mononuclear cells from individuals with stable atopic asthma. AB - Activated mononuclear cells (lymphocytes and monocytes) (MNCs) circulate in peripheral blood and accumulate in the airways of individuals with steady-state asthma. The expression of adhesion molecules on MNCs of 10 patients with steady state atopic asthma and 10 non-atopic control subjects was measured by flow cytometry. The mean fluorescence intensity of CD23 (P = 0.0003), CD11a (P = 0.034), and very late antigen-4 (VLA-4) (P = 0.016) was increased on lymphocytes of asthmatic patients relative to those of controls. Although the expression of CD16 (P = 0.002) and CD23 (P = 0.002), which are associated with differentiation into macrophages, was increased on monocytes of patients relative to those of controls, monocyte expression of VLA-4 (P = 0.006) and sialyl Lewis x (P = 0.005) was reduced. The concentration of interleukin-4 (IL-4) in serum was significantly higher in asthmatic patients than in normal subjects (P = 0.023), and a significant correlation was apparent between the serum concentration of IL-4 and the expression of VLA-4 on lymphocytes (p = 0.71, P = 0.03) or on monocytes (p = 0.72, P = 0.03) in asthmatic patients. Results suggest that IL-4 may contribute to the priming of adhesion molecule expression on lymphocytes even in steady state conditions in individuals with chronic allergic inflammation. PMID- 9208188 TI - Myeloperoxidase and interleukin-8 levels in chronic sinusitis. AB - We have recently phenotyped inflammation in non-infectious allergic and non allergic chronic maxillary sinusitis using sinus biopsies and lavage fluids. In this first paper, we have concentrated our work on the eosinophil, T cell, mast cell and macrophage infiltrates. However, many unresolved questions remain and particularly the role of neutrophils needed to be addressed. In the present study, we focused on the neutrophilic inflammation: myeloperoxidase (MPO) and interleukin-8 (IL-8) were measured by immunoassays and neutrophils were enumerated by conventional staining in the sinus lavage fluids of 16 patients with chronic sinusitis and six control subjects. Both MPO and IL-8 levels were significantly higher in patients than in controls (P < 0.01 and 0.005, respectively). There was a significant correlation between MPO levels and neutrophil numbers, and between MPO and IL-8 levels in the sinus lavage fluid (P < 0.0001, Spearman rank correlation). The presence of high levels of IL-8 in the lavage fluids of patients suffering from chronic sinusitis, levels which correlate with those of MPO, suggests that this cytokine may activate neutrophils in this chronic disease. PMID- 9208189 TI - Correlation between IgA antibody and eosinophil cationic protein levels in induced sputum from asthmatic patients. AB - BACKGROUND: Eosinophils are known to be main effector cells in allergic inflammation and IgA antibody has been shown to be a potent stimulus for eosinophil degranulation in in vitro conditions. OBJECTIVE: To evaluate the possible role of IgA antibodies on eosinophil degranulation in lower respiratory mucosa of asthmatics, we tried to find a correlation between total IgA and eosinophil cationic protein (ECP) levels in induced sputum from asthmatics. METHODS: We measured total IgA and albumin levels by nephelometry, and eosinophil cationic protein levels by Pharmacia CAP system in induced sputum from 23 atopic asthmatics and 12 healthy controls. RESULTS: IgA and albumin levels in induced sputum from asthmatics with sputum eosinophilia (sputum eosinophil count > or = 5% of 200 counted non-squamous cells) were significantly higher (P < 0.05) than those from controls. However, IgA and albumin levels in induced sputum from asthmatics without sputum eosinophilia were not significantly different with those from controls (P > 0.05). In induced sputum from asthmatics, ECP levels were significantly correlated with albumin (r = 0.44, P = 0.04) and IgA levels (r = 0.67, P = 0.002). ECP/albumin ratio was also significantly correlated with IgA/ albumin ratio (r = 0.61, P = 0.004). CONCLUSION: Our results support the hypothesis that IgA antibodies in tracheobronchial secretion may be involved in eosinophil degranulation in asthma, and further study is needed to prove this hypothesis. PMID- 9208190 TI - Molecular cloning and expression of a Penicillium citrinum allergen with sequence homology and antigenic crossreactivity to a hsp 70 human heat shock protein. AB - BACKGROUND: Penicillium citrinum has been identified as the most prevalent airborne Penicillium species in the Taipei area. However, detailed studies on allergens of this ubiquitous Penicillium species are still lacking. OBJECTIVE: For the characterization of allergens of this prevalent Penicillium species, molecular cloning and expression of the allergen genes of P. citrinum were performed in the present study. METHODS: Molecular cloning of the allergen genes was performed by using a lambda Uni-Zap XR cDNA library of P. citrinum and serum from an asthmatic patient. The cloned cDNA was excised from the phage vector as a recombinant pBluescript phagemid and sequenced. The cDNA of the IgE-binding clone was expressed as fusion protein with the glutathione-S-transferase. The corresponding natural allergen was analysed by absorption immunoblotting using monoclonal antibody and serum from asthmatic patient. The frequency of IgE binding to the allergen cloned was analysed by dot immunoassay using recombinant allergen and by immunoblotting using the whole extract of P. citrinum. RESULTS: In the screening of cDNA library of P. citrinum using serum from an asthmatic patient, IgE-binding cDNA clones designated SC4 and XL were obtained. The 5' truncated, 0.7-kb and 1.7-kb cDNA inserts of clones SC4 and XL contained open reading frames of 163 and 503 amino acids, respectively. On alignment, the deduced amino acid sequences showed that 97 (60%) of the 163 amino acids and 376 (75%) of the 503 amino acids were identical to the corresponding amino acid sequence of the human heat shock protein in the hsp70 family. Both recombinant SC4 and XL showed positive SDS-PAGE-immunoblot reactivity to a monoclonal antibody MA3-006 against the human hsp 70 protein. For characterization of the corresponding natural allergen, immunoblotting reactivities of MA3-006 and IgE antibodies to the 70 kDa component of P. citrinum have been shown to be disappeared after absorption of these antibodies with the recombinant SC4 protein. Sera from 14 (41%) of 34 Penicillium-allergic patients showed IgE binding to the recombinant XL protein and the 70 kDa component in the extract of P. citrinum. CONCLUSION: Results obtained suggest that hsp 70 is an allergen of P. citrinum and that clones SC4 and XL contain partial cDNAs of this allergen gene. PMID- 9208191 TI - Allergenic crossreactivity between Lepidoglyphus destructor and Blomia tropicalis. AB - BACKGROUND: In general, the non-pyroglyphid mites Lepidoglyphus destructor and Blomia tropicalis show a different geographical distribution. Allergic sensitization to both species have been demonstrated in several investigations. However, whether this reflects cross-reactivity or dual sensitization is so far not known. OBJECTIVE: The aim of the study was to investigate the allergenicity and allergenic crossreactivity of L. destructor and B. tropicalis using sera from Sweden and Brazil. METHODS: Allergens in extracts of L. destructor and B. tropicalis were identified with SDS-PAGE and immunoblotting and the crossreactivity was studied by an immunoblot inhibition method. In addition to mite extracts, a recombinant major allergen of L. destructor, Lep d 2, was used. RESULTS: The extract prepared from L. destructor contained 21 IgE-binding components when using the Swedish or the Brazilian sera. A 15 kDa allergen was recognized by 85% of the Swedish sera and 78% of the Brazilian. The B. tropicalis extract exposed 23 IgE-binding components when the Brazilian sera were used and 19 when the Swedish sera were used. A total of 83% of the Brazilian sera and 80% of the Swedish sera identified a 14.5 kDa allergen. The IgE response of the Swedish serum pool to 10 B. tropicalis allergens was inhibited by L. destructor extract. Likewise, the response of the Brazilian serum pool to four different L. destructor allergens was inhibited by B. tropicalis extract. The recombinant Lep d 2 allergen inhibited 33% of the IgE binding of the Swedish serum pool to the 14.5 kDa allergen in the B. tropicalis extract. CONCLUSION: Crossreactivity with several proteins from L. destructor and B. tropicalis was demonstrated. The results suggest that a B. tropicalis 14.5 kDa allergen is antigenically crossreactive with recombinant L. destructor allergen Lep d 2. PMID- 9208192 TI - Purification and IgE binding capacity of Der s 3, a major allergen from Dermatophagoides siboney. AB - BACKGROUND: Sensitization to the house dust mite Dermatophagoides siboney has been demonstrated in asthmatic patients. Previously, Dermatophagoides siboney group 1 and group 2 allergens, named Der s 1 and Der s 2, respectively, have been purified. OBJECTIVES: The aim of this study was to purify and to study the IgE reactivity of 30 kDa component, suspected to correspond to group 3 allergens. METHODS: The protein was purified by affinity chromatography using anti-Der f 3 monoclonal antibodies and semi-preparative SDS-PAGE. The IgE binding capacity of the purified fractions was tested with sera from 106 mite-sensitive asthmatic patients using a modified chemiluminiscent method. RESULTS: Affinity chromatography resulted in fractions containing the 30 kDa component which was further purified to homogeneity by SDS-PAGE. Seventy-three per cent of the sera showed IgE reactivity to this protein, indicating that it is a major allergen. The protein also reacted with anti Der f 3 polyclonal antibodies and had tryptic activity. There were differences in the reactivity to Der s 3 according to the age of the patients. CONCLUSION: Based on the frequency of IgE reactions and the reactivity with antibodies directed to Der f 3, it is proposed to name this 30 kDa allergen from D. siboney, Der s 3. PMID- 9208193 TI - Effects of JTV-506, a new K+ channel activator, on airway smooth muscle contraction and systemic blood pressure. AB - BACKGROUND: ATP-sensitive K+ (KATP) channel activators produce relaxation of smooth muscle in many tissues. However, this wide range of effects restricts their clinical usefulness in bronchial asthma because of a reduction in systemic blood pressure. METHODS: We have now examined the effects of JTV-506, a new benzopyran derivative, on airway smooth muscle contraction and systemic blood pressure and have compared this compound with cromakalim. We measured isometric tension records from guinea-pig isolated trachea, as well as the respiratory resistance (Rrs) and systemic blood pressure in anesthetized guinea-pigs. RESULTS: JTV-506 caused a concentration-dependent inhibition of histamine-induced contraction in guinea-pig isolated tracheal smooth muscle, and was antagonized by glibenclamide. JTV-506 was 7.6-fold more potent than cromakalim. In anesthetized animals the intravenous injection of JTV-506 reduced the increase in Rrs induced by intravenous application of 5 micrograms/kg of histamine in a dose-dependent manner. 10 micrograms/kg of JTV-506 resulted in 57.0 +/- 17.9% inhibition of the increase in Rrs at 10 min. The inhibitory action on Rrs disappeared after 60 min. 10 micrograms/kg of cromakalim caused 25.4 +/- 5.8% inhibition of the increase in Rrs induced by histamine at 1 min. The ED50 values for JTV-506 and cromakalim were 6.7 +/- 3.5 micrograms/kg and 60.1 +/- 15.8 micrograms/kg, respectively (P < 0.05). Cromakalim was approximately 9-fold less potent in inhibiting the increased Rrs by histamine, and the inhibitory action lasted less than 10 min. The reduction of systemic blood pressure by JTV-506 and cromakalim (each at a dose of 10 micrograms/kg iv) was 11.3% and 21.5%, respectively (P < 0.05). CONCLUSION: JTV-506 inhibits histamine-induced contraction of tracheal smooth muscle by activation of KATP channels. This compound is more potent and longer lasting in the suppression of histamine-induced increases in Rrs, and is less hypotensive than cromakalim. Our results suggest that this compound merits further investigation for utility as a bronchodilator in the clinic. PMID- 9208194 TI - Review: the biochemistry of common aeroallergens. C. A. Stewart and P. J. Thompson, September 1996; 26:1020-44. PMID- 9208195 TI - Virus zoonoses--a long-term overview. PMID- 9208196 TI - Prevention of Salmonella typhimurium caecal colonisation by different preparations for competitive exclusion. AB - The number of Salmonella typhimurium per gram of caecal contents of chicks, given orally prior to infection with different preparations containing protective intestinal flora, was significantly lower than in the control group. The best results were obtained when the lyophilised caecal rinses were given orally to 1 day-old chicks. Less effective methods of protection of the alimentary tract proved to be oral application to chicks of fresh faeces or lyophilised faeces and administration of the lyophilised caecal rinses with drinking water. Oral application of undefined anaerobic caecal culture to chicks proved to be the least effective of the methods investigated of protecting them against Salmonella colonisation, although even these results differed statistically from those of the controls. The percentage of infected birds 7 days after challenge confirmed the efficacy of oral application of lyophilised caecal rinses--only 30% of individuals from the group treated in this way proved to be Salmonella positive. In contrast, the percentage of infected birds in other experimental groups ranged from 50% to 75% and was 100% in the controls. PMID- 9208197 TI - Bovine leukemia virus: early reflections in blood after an experimental infection of calves. AB - In order to study early alterations in the blood following infection with bovine leukemia virus (BLV) in the natural host, 15 calves were inoculated with blood from a BLV-positive donor cow. The humoral immunological response was followed by ELISA for 2 months. Seroconversion to BLV was demonstrated at 4-5 weeks post infection. Total and differential leukocyte counts were performed. Acute lymphocytosis was observed at the time of seroconversion in the majority of the experimental calves. By the aid of monoclonal antibodies (mAbs), the proportion as well as the total number of lymphoid cells were studied in four of the calves, applying analytical flow cytometry. At the time of seroconversion the percentage of B-cells increased from 19.1 +/- 7.5% to 37.9 +/- 15.8%, and the T-cells (CD2+) decreased from 36.7 +/- 7.3% to 22.7 +/- 6.0%, the latter being attributable to decreases in the percentage of CD4+ as well as CD8+ T-cells for the infected calves together. Subsequently, altered B/T ratios were observed. In one of the calves an increase in the absolute number of CD5+ cells coincided with an increase in total B-cells. The early phenotypic alterations in lymphocyte subsets, before and after seroconversion to BLV, were comparable to those of non lymphocytotic and persistent lymphocytotic cattle, respectively. Sera from 15 calves were tested for the presence of interferon (IFN), as measured by antiviral activity. BLV does not appear to induce the production of IFN. PMID- 9208198 TI - Toxoplasmosis: comparative species susceptibility and host immune response. AB - The protozoan parasite Toxoplasma gondii is capable of infecting all warm blooded animals; however, the consequences of infection are very variable between different species of animal. Marsupials and New World monkeys, which have evolved largely separately from the cat, the definitive host of the parasite, are among the most vulnerable species where infection with T. gondii can prove fatal. In more resistant species such as humans and sheep, infection is generally unapparent, provoking only mild symptoms; thereafter the host remains infected for life. However, when the immune system is compromised, such as in the immunologically immature fetus, infection with the parasite can have very serious consequences. Much of the work examining host immune responses has been done using experimentally infected mice. While there are many advantages in using this experimental model, care should be taken in extrapolating results from mice to other species. Mice are extremely vulnerable to the consequences of infection with T. gondii., and their use to further our understanding of congenital toxoplasmosis may not be ideal, as fetal infection can occur in successive pregnancies. This is not the case in rats or sheep; they are more resistant to the disease and therefore may provide a more relevant model for human congenital toxoplasmosis. Studies of host immune responses have emphasised the importance of the cytokine interferon gamma (IFN gamma) in resistance to T. gondii. The efficiency of induction of this cytokine may be critical for determining the outcome of the host-parasite relationship. PMID- 9208199 TI - Poxvirus preparation CONPIND initiates production of the major inflammatory mediators IL-1 alpha and TNF-alpha in human whole blood and in blood mononuclear cell cultures. AB - The poxvirus preparation CONPIND initiated the production of the two major inflammatory mediators interleukin-1 alpha (IL-1 alpha) and tumour necrosis factor-alpha (TNF-alpha) in whole blood and in blood mononuclear cell cultures of man at 37 degrees C within 12-48 h. The release of the cytokines followed a dose efficacy curve. High levels of IL1-alpha (up to 810 pg/ml) and TNF-alpha (up to 533 pg/ml) were determined by sandwich enzyme-linked immunosorbent assay. PMID- 9208200 TI - Role of immune responses to a GroEL heat shock protein in preventing brucellosis in mice vaccinated with Brucella abortus strain RB51. AB - Resistance to infection with virulent Brucella abortus strain 2308 and antibody and lymphocyte proliferative responses to a recombinant 60 kDa B. abortus GroEL heat shock protein were measured in mice vaccinated with attenuated B. abortus strain RB51. Mice at 12-20 weeks after vaccination with 5 x 10(8) colony forming units (CFU) of strain RB51 had increased resistance to infection with strain 2308 and increased antibody and lymphocyte proliferative responses to GroEL following challenge infection with 2308. However, these mice at 12-20 weeks after vaccination did not have greater resistance to infection than mice vaccinated with 5 x 10(6) CFU of strain RB51, which had no increased antibody or lymphocyte proliferative response to GroEL. These results indicate that mice vaccinated with strain RB51 can have antibody and cell-mediated immune responses to GroEL during infection with virulent strain 2308, although neither response appeared to have an essential role in vaccine-induced immunity to brucellosis. PMID- 9208201 TI - Comparative effects of bovine cytokines on cattle and bison peripheral blood mononuclear cell proliferation. AB - Proliferation of peripheral blood mononuclear cells (PBMC) from cattle and bison was measured following stimulation of PBMC with bovine cytokines. Bovine interleukin 1 beta (BoIL-1 beta), interleukin 2 (BoIL-2) or granulocyte macrophage colony-stimulating factor (BoGM-CSF) at 0.1-100 U/ml were incubated for 48 h with PBMC alone or with PBMC and various mitogens. These included concanavalin A (Con A), phytohemagglutinin (PHA), pokeweed mitogen (PWM) or Escherichia coli 055:B5 lipopolysaccharide (LPS) at 10-0.1 micrograms/ml. BoIL-2 alone, but not BoIL-1 beta and BoGM-CSF alone, induced proliferation of cattle and bison PBMC in the absence of mitogens. In addition, BoIL-1 beta and BoIL-2, but not BoGM-CSF, enhanced proliferation of cattle and bison PBMC induced by mitogens. These results indicate that BoIL-1 beta and BoIL-2 stimulate cattle and bison PBMC proliferation in a similar manner, whereas BoGM-CSF does not appear capable of stimulating either cattle or bison PBMC proliferation. PMID- 9208203 TI - Interference activity of bovine herpes virus 1 strains against Aujeszky's disease virus in cell cultures. AB - The interference-inducing activity of different low- and high-virulence strains of bovine herpes virus 1 (BHV-1) against Aujeszky's disease virus was studied by a parallel titration on permissive and non-permissive cell culture. The low virulence BHV-1 strains possessed better interference-inducing activity than the high-virulence strains. An interference blocking test (IBT) was developed for the detection of BHV-1 antibodies in bovine sera. Comparative results of IBT and the virus neutralisation reaction did not show statistically reliable differences. PMID- 9208202 TI - Non-O157 Vero cytotoxin producing Escherichia coli: aetiological agents of diarrhoea in children in Dunedin, New Zealand. AB - Strains of Escherichia coli that produce Vero cytotoxin (VTEC) commonly cause diarrhoea, haemorrhagic colitis and haemolytic-uraemic syndrome in many northern hemisphere countries. In these countries, serotype O157:H7/H-predominates and has caused large food-borne outbreaks of infection. In contrast, few cases of infection with this serotype have been reported in New Zealand. Over a 3-month period, 484 stool specimens submitted to medical laboratories in Dunedin were screened for E. coli O157:H7/H-using sorbitol MacConkey agar, Y1 and Vero cell assays. Where possible, Vero cytotoxin production was confirmed by an ELISA test. Specimens from children aged 12 years or less were additionally screened for non O157 VTEC. In the specimens of the children tested, O157:H7/H-VTEC was not isolated, but VTEC belonging to other serogroups were isolated from the children. Of interest was the detection of other species of Enterobacteriaceae, which produced a cytopathic effect on Vero cells. This study confirms the low incidence of infection with O157:H7/H- VTEC in New Zealand and suggests that non-O157 VTEC is a more important cause of diarrhoeal disease. PMID- 9208204 TI - Antigenic epitope characterization of matrix protein of Newcastle disease virus using monoclonal antibody approach: contrasting variability amongst NDV strains. AB - A panel of 15 monoclonal antibodies (MABs) against matrix (M) protein of Newcastle disease virus (NDV) was obtained and the specificity towards the M protein was proven by radioimmunoprecipitation assay and antigen capture enzyme linked immunosorbent assay (ELISA). Further studies were directed to antigenic epitope mapping of the M protein by means of this panel. The epitope characterization was performed by competitive antibody-binding assay by means of labelling each MAB with biotin [3]. At least three clear non-overlapping and two partially overlapping groups were determined, each including four, one, eight, one, and one MAB, respectively. All the above MABs appeared to be induced by structural epitopes formed in conditions of tertiary structure of the native M antigen. Twelve reference and 51 recently isolated local NDV strains have been studied by means of this MAB panel, several lineages having been revealed. The high stability of some epitopes and different variability of the others was demonstrated. No correlation between the above lineages and some other properties of the studied NDV strains (host specificity, date and place of isolation) has been found. PMID- 9208205 TI - Toxoplasma gondii infection in sheep and cattle. AB - Toxoplasma gondii is a protozoan parasite that can infect all warm-blooded animals. Sheep and cattle show different susceptibilities to T. gondii infection. Primary infection in pregnant sheep can result in abortion or the birth of weak lambs but they are then protected against further challenge by the development of an effective immunity. Cattle on the other hand can be readily infected, but abortion or perinatal mortality have not been recorded. The evidence suggests that cattle develop a more effective immune response to T. gondii infection than sheep. Potential mechanisms to explain these differences are discussed in this paper. PMID- 9208206 TI - Regulation of neuronal nitric oxide synthase through alternative transcripts. AB - Nitric oxide (NO) participates in diverse physiological processes ranging from neurotransmission to muscle relaxation. Neuronal-derived NO can be either beneficial or detrimental depending on the cellular context. Neuronal NO synthase (nNOS) must therefore be tightly regulated. One level of regulation involves synthesis of numerous nNOS mRNA transcripts. At least six distinct molecular species of nNOS mRNA are expressed in a tissue and developmentally-regulated manner. Alternative splicing allows the creation of nNOS proteins differing in both enzymatic characteristics and structural features. As one example, we find that there are nNOS mRNAs lacking exon 2. One isoform, nNOS beta, retains full enzymatic activity but lacks a major protein-protein interaction domain (PDZ domain) responsible for targeting nNOS to synaptic membranes. This alternative splicing produces a mislocalized but fully active protein which may be relevant to certain pathologies. As evidence of this, we find that many human brain tumors express an alternatively spliced form of nNOS that co-migrates with nNOS beta, and lacks exon 2. Finally, we also find a 2.5-kb testis-specific nNOS mRNA corresponding to the C-terminal reductase domain of nNOS whose function is unclear. PMID- 9208207 TI - Increased expression of neuronal nitric oxide synthase in bladder afferent and spinal neurons following spinal cord injury. AB - Changes in the distribution of neuronal nitric oxide synthase immunoreactivity (NOS-IR) after chronic (5-6 weeks) spinal cord injury (SCI) were examined in bladder afferent and spinal neurons in the region of the sacral parasympathetic nucleus (SPN) in the L6-S1 spinal segments. Bladder afferent neurons in the L1, L2, L6 and S1 dorsal root ganglia (DRG) were identified by retrograde axonal transport following injection of fluorogold (FG) into the urinary bladder. A differential distribution of NOS-IR was detected in DRG cells at different segmental levels of spinal cord intact animals with significantly greater numbers of NOS-IR cells present in thoracic (T8, T12; 30-40 NOS-IR cell profiles/section) and rostral lumbar (L1) DRGs (18 NOS-IR cell profiles/ section) compared to caudal lumbosacral (L5-S1) DRGs (0.2-0.4 NOS-IR cell profiles/section). A significant increase in the number of NOS-IR cells was detected in the L6-S1 DRG (p < or = 0.001; 12-20 NOS-IR cell profiles/section) and in the L1-L2 DRG (15-40 NOS-IR cell profiles/section) but not in the L5 DRG following SCI. In these ganglia, an average of 41.2 +/- 7.8% (L6) and 36.3 +/- 0.9% (S1) of FG-labeled bladder afferent neurons were NOS-IR. In contrast, in spinal cord intact animals, no FG-labeled bladder afferent neurons were NOS-IR. Following SCI, NOS-IR fibers were detected along the lateral edge of the dorsal horn extending from Lissauer's tract to the region of the SPN (lateral collateral pathway of Lissauer) of the L6 and S1 spinal segments. These NOS-IR fibers were not detected in adjacent spinal segments (L5, S2). SCI also significantly (p < or = 0.001) increased the number of spinal neurons in the region of the SPN (presumptive preganglionic neurons) in the L6-S1 spinal segments exhibiting NOS-IR. These results indicate that NOS-IR in bladder afferent and spinal neurons is plastic and can be up-regulated by chronic SCI. Changes in the neurochemical properties of these neurons after SCI may be mediated by pathological changes in the target organ (i.e., urinary bladder) and/or spinal cord. PMID- 9208208 TI - Regulation of nitric oxide synthase expression in motoneurons following nerve injury. AB - This study investigated the influence of postsynaptic targets and axonal ensheathing cells on the expression of nitric oxide synthase (NOS) in axotomized neurons. The hypoglossal and vagus nerves of adult rats were lesioned unilaterally, and axotomized neurons expressing NOS, identified by NADPH diaphorase histochemistry and NOS immunocytochemistry, were counted as a function of time between 1 and 70 days postaxotomy. In the dorsal motor nucleus of the vagus (DMV), the number of NOS-positive neurons increased steadily with a time course which was not significantly different between the nerve crushed and transected groups. In the hypoglossal nucleus, each of four axotomy groups, namely, crushed, transected, ligated/transected, and avulsed, exhibited a distinct time course and extent of NOS expression, suggesting that postsynaptic targets and axonal ensheathing cells regulated NOS expression. The disparity between hypoglossal and DMV neurons in NOS expression may be due to the latters' unresponsiveness to, or inability to obtain the proper regulatory signals. Since cell loss was most severe in the hypoglossal nucleus following nerve avulsion, it appears that prolonged expression of NOS at high levels may be neurotoxic. PMID- 9208209 TI - The effects of remaining axons on motoneuron survival and NOS expression following axotomy in the adult rat. AB - it is well known that target removal or distal axotomy in adult animals results in no detectable loss of motoneurons in the spinal cord. By performing axotomy in the seventh cervical (C7) spinal nerve at different distances from the spinal cord (0, 2, 4, 8 mm respectively), this study examines the effects of the remaining axons on motoneuron survival as well as NOS expression. Results of the present study show that axotomy in adult peripheral nerve can induce significant spinal motoneuron death if axotomy is performed close enough to the spinal cord. The closer the axotomy to the spinal cord, the higher the rate of motoneuron loss was observed. The most significant motoneuron loss was found in animals with axotomy at 0 mm to the cord, which was coincident with the highest percent of NOS positive motoneurons. The rate of survival of motoneurons increases and the percent of NOS-positive motoneurons decreases when the distance of the axotomy to the cord increases from 0 to 4 mm. No significant motoneuron loss nor NOS positive motoneurons were observed when axotomy was performed at 4 mm and distally. These results indicate that the survival of spinal motoneurons in adult rat following axotomy is largely dependent on the length of the remaining axons. The longer the remaining axon, the better for motoneuron survival. The minimal length of axon for motoneuron survival in adult rat seems to be at least 4 mm. PMID- 9208210 TI - Inhibition of nitric oxide synthase fails to disrupt the development of cholinergic fiber patches in the rat superior colliculus. AB - Nitric oxide may serve as a retrograde messenger to refine or stabilize synapses in the developing nervous system. Whether this action is dependent upon glutamate and the N-methyl-D-aspartate receptor is not yet established. We have used the patch-cluster system in the intermediate gray layer (IGL) of the rat superior colliculus (SC), a system receiving both glutamatergic and cholinergic input, to study this question. The normal distribution and development of nitric oxide synthase (NOS) in SC was examined using nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) histochemistry in Sprague-Dawley rats aged P4 to adulthood. Fibers containing acetylcholine (ACh) were identified using choline acetyltransferase (ChAT) immunocytochemistry. In addition, N omega-nitro-L arginine, an inhibitor of NOS, was injected intraperitoneally from birth until P10, P14, P18, or P21-22 to determine if NOS inhibition would disrupt the formation of the ACh patches. Control animals were studied from the same age groups. Our results show NADPH-d-labeled cells within the periaqueductal gray and the deep gray layer of SC by P4, the earliest age examined. By P8-P9, cells in the IGL were well labeled by NADPH-d, while few in the superficial layers (SL) were labeled. SL cells were visible by P10 and were intensely labeled by P14. IGL cells transiently expressed NADPH-d in that the number of labeled cells increased from P8 to P35, then decreased in the adult. ChAT-labeled fibers first appeared in the IGL at P10, formed a characteristic two-tier pattern by P14, and established obvious patches by P21. Inhibition of NOS from birth produced no qualitative differences in the distribution or density of either ChAT-labeled fibers or NADPH-d-labeled cells and fibers at any of the ages examined. We therefore conclude that NO does not contribute to the refinement of cholinergic fiber patches in the rat SC, probably because the fiber system is not glutamatergic. PMID- 9208211 TI - Cerebellar nitric oxide synthase is expressed within granule cell patches innervated by specific mossy fiber terminals: a developmental profile. AB - The nicotinamide adenine dinucleotide phosphate diaphorase (NADPH-d) staining technique was utilized as a marker of nitric oxide synthase (NOS) to map NOS expression in developing and adult rat cerebellum. NADPH-d-positive cells were first visualized in the cerebellar cortex at postnatal day 5 (PND5) which increased to peak levels by PND30 when they began to exhibit a patch-like organization. In order to determine the relationship of the NADPH-d staining pattern with mossy fiber innervation, mossy fiber projections were traced using cholera toxin B subunit or biocytin injected into the lateral reticular nuclei (LRtN) or pontine nuclei (PtN), respectively. Double staining revealed that the clustered mossy fiber terminals projecting from the ventrorostral LRtN and caudal PtN were well matched with NADPH-d-stained patches. This patch-like localization of NOS matched with specific mossy fiber terminals in adult cerebellum implicates these NOS patches as defining distinct anatomical zones. PMID- 9208212 TI - The developmental expression of neuronal nitric oxide synthase in cerebellar granule cells is sensitive to GABA and neurotrophins. AB - In the adult cerebellar cortex, the differential expression of neuronal nitric oxide synthase (nNOS) can be used to discern discrete subsets of granule neurons. These subsets originate concurrently with the afferent innervation of granule neurons. Using primary cultures established from murine cerebella at different development stages, we analyzed the effects of the neurotransmitter GABA and the neurotrophins BDNF and NT-3 on nNOS expression in developing granule cells. We show a biophasic effect of GABA on nNOS expression, which correlates with the ability of this transmitter to increase intracellular calcium levels. BDNF and NT 3 are shown to increase nNOS levels in cultures derived from postnatal, but not in those from embryonic cerebella. Together with previous findings [Baader and Schilling; J Neurosci 1996; 16:1440-1449], these data suggest a scenario in which afferent innervation and growth factors cooperate to differentially affect nNOS expression at discrete developmental stages of cerebellar granule cells. PMID- 9208213 TI - Simulation of the influence of different factors on the motor nerve conduction velocity measurement. Part 2: Pathological conditions. PMID- 9208214 TI - Distal muscular dystrophy. Case reports. AB - The distal muscular dystrophy has been described in 1902 by Gowers. Since then a lot of cases with different mode of inheritance, clinical involvement and morphological findings have been described. Obviously distal myopathies are not a single entity. We describe four new cases of distal myopathy. Although we suppose that they are from the adult-onset Welander's form, they are different in some points. Two cases were with onset in the lower limbs and autosomal dominant inheritance, the third one was sporadic with onset in the lower limbs and the last one was sporadic with onset in the upper limbs. The distal muscles were markedly, while the proximal were slightly involved. Sternocleidomastoid, neck or facial muscles wasting were also found in three cases. The serum creatine kinase in all patients was in nearly normal limits. The electromyographic findings were consistent with chronic myopathy, although fibrillation potentials and positive sharp waves were found in three cases. Muscle biopsy data confirm the myopathic involvement in all patients. We have described patients, which confirm the variability of distal myopathies, regarding the age of onset, initial symptoms, clinical picture, electromyographic and muscle biopsy findings. PMID- 9208215 TI - Nuclear facial palsy in multiple sclerosis: a case report. AB - We encountered an unusual case of isolated subacute nuclear palsy of the facial nerve in a 31-year-old man with a 3-year history of clinically established relapsing form of multiple sclerosis. Typical positive sharp waves and fibrillation potentials in the affected facial muscles detected by means of electromyography showed that the lesion was located in the lower motor neuron. Transcranial magnetic stimulation of the cortex could not evoke normal compound motor unit potentials, however, transcranial magnetic stimulation of the facial nerve, which is strongly supposed to depolarize the peripheral nerve in the area of the root exit zone near the internal acoustic meatus, elicited an adequate response. Taking these findings as well as the results of positive magnetic resonance imaging into consideration, we concluded that the clinical symptom was due to a plaque of demyelinization in the nucleus of the facial nerve or in the proximal intramedullary course of facial fibres. PMID- 9208216 TI - The evaluation of mannitol therapy in acute ischemic stroke patients by serial somatosensory evoked potentials. AB - The effects of mannitol infusion therapy on acute ischemic stroke were investigated by serial median nerve somatosensory evoked potentials (SEP) in 31 patients with abnormal SEP recordings within 72 hours after onset. In 10 patients with missing N20 waves mannitol had no effect. In 12 patients mannitol improved N20 and central conduction time (CCT) latencies. In 9 patients absent N20 waves appeared after mannitol. As regard to the location of infarcts, CT revealed 7 hemispheric infarcts in patients with absent N20 waves in spite of mannitol infusion. The remaining patients' CTs revealed cortical and subcortical infarcts. This study concludes that mannitol treatment improved microcirculatory flow in the ischemic penumbra of acute ischemic stroke patients. We suggest that SEPs can be used in evaluating the effects of drugs to be used in the therapy of acute cerebrovascular accidents. PMID- 9208217 TI - Sensory nerve recovery following median nerve provocation in carpal tunnel syndrome. AB - The purpose of this study was to evaluate the recovery of median nerve sensory nerve action potentials (SNAP) following median nerve provocation in hands with carpal tunnel syndrome (CTS). Repeated nerve conduction measurements were performed before and after wrist flexion combined with resisted finger flexion in 35 hands with a clinical diagnosis of CTS and in 25 asymptomatic control hands. Orthodromic sensory median nerve potentials were recorded over an 8 cm segment between the palm and wrist. Hands with CTS had significant reductions in nerve potential amplitude and latency following median nerve provocation. Hands with mild to moderate CTS, had the greatest reductions in nerve potential amplitude and the longest amplitude recovery times following median nerve provocation. Determination of changes in amplitude and amplitude recovery time of the median nerve SNAP following median nerve provocation has the potential to improve the accuracy of the electrophysiological diagnosis of CTS. PMID- 9208218 TI - Muscle surface mechanical and electrical activities in myotonic dystrophy. AB - The dimensional changes of the muscle fibres of the active motor units generate a signal, labelled as mechanomyogram (MMG), related to the number and firing rate of the active fibres. Aim of the study was to evaluate if the root mean square (RMS) of the surface EMG, of the MMG and their relationship (electromechanical coupling efficiency, EMCE) are related to the clinical stage of patients affected by myotonic dystrophy. The EMG and the MMG were recorded during isometric contractions at 20%, 40% and 60% of the maximal voluntary contraction (MVC), in less (elbow flexors, EF) and more (finger flexors, FF) affected muscles in 10 myotonic dystrophy (DM) patients (duration of disease: 15-22 yr.; MDRS scale: 3 4) and in 10 age matched controls (C). It resulted that I) the MVC was only 30% lower in DM than in C (P > 0.05) for the EF but 74% lower (P < 0.001) for the FF; II) at the same relative force the MMG-RMS and the EMG-RMS in DM were significantly lower than in C (P < 0.05); III) the EMCE was almost the same in DM and C for EF but dramatically lower for FF. In conclusion these findings provide evidence that the EMCE changes due to the dystrophic process can be monitored by this new and non invasive technique. A good concurrence of clinical findings and experimental results was observed. PMID- 9208219 TI - A study on new diagnostic criteria of H reflex. AB - H reflex is known as a useful electrodiagnostic test in the diagnosis of S1 radiculopathy. But, only the latency difference has been the useful parameter by previously published conventional method. Under the assumption that the constant appearance of initial negative biphasic H wave is critical to study H reflex using parameters such as amplitude, area and shape, we developed a new method using parameters such as amplitude, area and shape. To validate our assumption and to compare the diagnostic values between the conventional method and the new one, we studies H reflex in 330 subjects. One hundred sixty-two subjects were studied by conventional method and 168 subjects were studied by our new method. There was no definite difference in diagnostic values between two methods by latency criteria. However, new method was more specific for S1 radiculopathy than conventional method by amplitude and area criteria. Parameters such as amplitude, area and shape can be used significantly only in the new method. Therefore, we suggest new diagnostic criteria of abnormal response as follows: (1) H latency difference over 1.0 msec and H/H amplitude ratio less than 0.5 or (2) H latency over 30 msec or (3) unilateral absent evoked H response. PMID- 9208220 TI - Compression of the peripheral branches of the sciatic nerve by lipoma. AB - The authors report two female patients with chronic sensitive and motor findings in lower limbs caused by compression of distal branches of sciatic nerve by lipoma. Similar cases were not described on literature. Nerve conduction studies allowed to localize the exact site of compression. At surgery, lipomas compressing the deep peroneal nerve (case 1) and the posterior tibial nerve (case 2) were observed. Histologic studies of tumors confirmed the diagnoses. PMID- 9208221 TI - The Sec20/Tip20p complex is involved in ER retrieval of dilysine-tagged proteins. AB - Sec20p and Tip20p were previously identified as two interacting proteins involved in early steps of the secretory pathway in Saccharomyces cerevisiae. Here we describe a novel temperature-sensitive allele of TIP20 and analyze its phenotype. While sec20 and tip20 mutants exhibited a defect in forward ER-to-Golgi transport at the non-permissive temperature, both were also defective for retrieval of various dilysine-tagged proteins from the Golgi back to the endoplasmic reticulum (ER) at lower temperature. Dilysine-dependent Golgi localization of Emp47p was also defective in both mutants. These results suggest a role for the Sec20/Tip20p complex in retrieval of dilysine-tagged proteins back to the ER. PMID- 9208222 TI - Endoplasmic reticulum retention mediated by the transmembrane domain of type II membrane proteins Sec12p and glucosidase 1. AB - The yeast Sec12p, a type II protein localized to the yeast endoplasmic reticulum (ER), is similarly localized to the ER when expressed in mammalian cells. Replacing the transmembrane domain of the plasma membrane molecule dipeptidyl peptidase IV (D4) with that of Sec12p or the ER-localized enzyme glucosidase 1 resulted in the ER retention of the chimeric molecules, as assessed by immunocytochemical localization and the persistence of pulse-labeled proteins in the endoglycosidase H-sensitive form. Retention is not due to gross misfolding as these chimeras remained enzymatically active. Density gradient analysis revealed that the ER-localized chimeric molecules form high molecular weight oligomers quickly after synthesis. The type II transmembrane domain of ER proteins could therefore mediate retention in the ER. PMID- 9208223 TI - Import of human and Thermoplasma 20S proteasomes into nuclei of HeLa cells requires functional NLS sequences. AB - Proteasomes are present both in the nucleus and cytoplasm of eukaryotic cells. Their localization is regulated and changes during the cell cycle. Nuclear localization signal (NLS) type sequences were identified in proteasomes from various organisms. In addition, acidic complementary sequences were identified (cNLS) which could interact with the positively charged NLS, masking or unmasking them and thereby modulating nuclear import. In this paper we show that fluorescently labeled human erythrocyte 20S proteasomes accumulate in the nucleus of digitonin-permeabilized cells. This translocation is ATP-dependent and occurs through the nuclear pore complex as is shown by blocking of the nuclear pores with wheat germ agglutinin. In addition, we used 20S proteasomes from Thermoplasma acidophilum as a model system. Recombinant 20S proteasomes from the archaebacterium Thermoplasma acidophilum are imported into nuclei of HeLa and 3T3 cells similar to their eukaryotic counterpart. We constructed mutants in the putative NLS and cNLS region to study their effect on import. The NLS mutant was not imported into nuclei and showed cytoplasmic staining only. This indicates that this sequence is indeed responsible for nuclear targeting. Mutational studies of the cNLS do not support the involvement of this sequence in regulation of nuclear transport. PMID- 9208224 TI - Sequential protein secretion from three distinct organelles of Toxoplasma gondii accompanies invasion of human fibroblasts. AB - Invasion of vertebrate cells by the protozoan Toxoplasma gondii is accompanied by regulated protein secretion from three distinct parasite organelles called micronemes, rhoptries, and dense granules. We have compared the kinetics of secretion from these different compartments during host cell invasion using immunofluorescence, immunoelectron microscopy, and quantitative immunoassays. Binding to the host cell triggered apical release of the micronemal protein MIC2 at the tight attachment zone that forms between the parasite and the host cell. In a second step, invagination of the host cell plasma membrane was initiated by discharge of the rhoptry protein ROP1 to form a nascent parasitophorous vacuole (PV). ROP1 was fully discharged into the vacuole by the time invasion was complete. In contrast to these very rapid early events, release of the dense granule markers GRA1 and NTPase was delayed until after the parasite was fully within the PV, eventually peaking at 20 min post-invasion. The sequential triggering of secretion from different organelles implies that their release is governed by separate signals and that their contents mediate distinct phases of intracellular parasitism. PMID- 9208225 TI - A kinesin-like mechanoenzyme from the zygomycete Syncephalastrum racemosum shares biochemical similarities with conventional kinesin from Neurospora crassa. AB - The kinesin superfamily consists of mechanoenzymes that convert chemical energy, stored in nucleoside triphosphates, into movement along microtubules. The founding member of this protein superfamily, the so-called conventional kinesin, was only known from animal sources until the recent description of kinesin from the filamentous fungus Neurospora crassa (G. Steinberg, M. Schliwa, Mol. Biol. Cell 6, 1605-1618 (1995)). To determine whether similar motors with comparable features are common in other filamentous fungi, a kinesin from a zygomycete, Syncephalastrum racemosum, was purified. Here, the isolation and characterization of this motorenzyme is described. The purified protein consisted of a doublet at 112 kDa and 115 kDa with no additional polypeptides. This was consistent with a calculated molecular mass of approximately 240 kDa and suggests that the motor is a dimer with a more globular shape than conventional kinesin from animal sources. In gliding assays the enzyme moved microtubules at 2.5 to 3.4 microns/s and had a nucleotide specificity similar to the Neurospora kinesin motor. Peptide antibodies against conserved regions in the head and the tall domain of conventional kinesins cross-reacted with the Syncephalastrum motor. In vitro, the enzyme was able to drive the microtubule-dependent movement of vesicles isolated from Syncephalastrum racemosum, as well as Neurospora crassa, and Aspergillus nidulans. In summary, the Syncephalastrum motor has many of the unique features in common with the conventional kinesin from the ascomycete Neurospora crassa and probably shares a similar function in living hyphae. PMID- 9208226 TI - Circular ruffle formation in rat basophilic leukemia cells in response to antigen stimulation. AB - Rat basophilic leukemia cells have previously been described to undergo striking cell surface changes after IgE-mediated stimulation of histamine secretion, whereby the dorsal surface loses its microvilli and acquires characteristic wavy ruffles. We have found using scanning electron microscopy, phase contrast and immunofluorescence, that a proportion of these cells also exhibit the formation of circular membrane ruffles on their dorsal surface after exposure to an IgE directed secretagogue; some cells also develop circular membrane ruffles following stimulation by phorbol myristate acetate or by calcium ionophore A23187. A flattened morphology appears to be linked to circular membrane ruffle formation in that these ruffles were found in areas of presumed cell spreading which are largely devoid of intermediate filaments and displaced to one side of the cell's nucleus, and they were not observed on rounded cells. This is in contrast to the wavy ruffles which are found on the entire cell surface including the region overlying the nucleus, and which are observed in rounded cells as well as spread cells. Circular ruffle formation and secretion are triggered by similar concentrations of antigen, but the circular ruffles are formed more slowly and only become abundant at times after most of the histamine has been released. The circular membrane ruffles showed no obvious association with endocytosis, as detected using fluorescein isothiocyanate-dextran as a fluid phase marker. The position of accumulation of endocytotic vesicles occurring subsequent to secretion was not found to be related to the circular membrane ruffles, but was observed around the nucleus. Circular membrane ruffles contain F-actin, and their formation is prevented by cytochalasin D. At least three types of myosin, types I, II and V are present and presumably play a role in circular ruffle formation. PMID- 9208227 TI - Processing of the gap junction protein connexin50 in the ocular lens is accomplished by calpain. AB - Gap junction channel forming connexins share a common membrane topology which has four transmembrane spanning segments with the amino- and carboxy termini both located on the cytoplasmic side. Both, mutation and truncation of the carboxyl tail of some connexins have been shown previously to have profound effects on channel function. Truncation of the carboxyl tail of connexin50 (Cx50) and connexin46 (Cx46) occurs naturally during the maturation of fiber cells in the mammalian lens. This system therefore offers the unique opportunity to study not only the cleavage process but also the functional role played by the cleaved domain, in a physiologically relevant context. As a first step, we now report on the cleavage of the 70 kDa ovine isoform of Cx50. The calcium-activated neutral protease calpain (EC 3.4.22.17) was identified as the enzyme which removed a 32 kDa carboxyl portion from the Cx50 molecule in mature lens fiber cells. The amino terminal 38 kDa portion remained embedded in the plasma membrane and was isolated and visualized as channel structures. The amino-terminal sequence of the cleaved 32 kDa portion matched an interior portion of the published amino acid sequence of the ovine Cx50 isoform. Thus, two closely spaced calpain cleavage sites were identified in the Cx50 molecule which were located carboxy-terminal from the predicted exit of the fourth transmembrane spanning segment by 62 or 72 amino acid residues, respectively. These data provide the basic information required for the future construction of Cx50 mutants to explore the functional consequences of this cleavage. PMID- 9208228 TI - Photoreceptor differentiation analyzed by the novel monoclonal antibody 1G1. AB - In vertebrates, photoreceptor development has become a key model system to study mechanisms of cell differentiation. A still unresolved question is why photoreceptor maturation is retarded over an extended period of embryogenesis though photoreceptors are among the first cells born in the retina. We have generated the novel monoclonal antibody 1G1 which binds to outer photoreceptor segments of adult retinae of various species including chicken and rat. In the developing chicken retina presumptive photoreceptor cells were labeled by MAb 1G1 at embryonic day 10 (E10). Retinal cell cultures revealed that the corresponding antigen is expressed on the cell surface of rods and cones likewise. Metabolic labeling with bromodeoxyuridine in vitro indicated that 1G1 antigen expression is restricted to postmitotic cells. Comparison of single cell cultures starting from different developmental stages showed that antigen expression can be induced prematurely, if cells are released from their native tissue environment. In order to analyze potential regulatory cell interactions, retinal cells were cultured on cryosections of the eye (cryoculture). The percentage of 1G1+ cells which contacted the pigment epithelium, was significantly lower in comparison to cells located on retinal tissue. The data are consistent with the notion that the pigment epithelial cells which contact retinal photoreceptors in vivo, could be partially inhibitory and consequently delay photoreceptor differentiation. PMID- 9208229 TI - Milk accumulation triggers apoptosis of mammary epithelial cells. AB - Continuous milk production is a consequence of a complex interplay of lactogenic hormones and it depends on the suckling stimulus during lactation. Involution is associated with a massive engorgement of the gland with milk followed by apoptosis of secretory epithelial cells and a restructing of the gland. Sealing of a single gland during lactation is sufficient to induce an initial engorgement and a subsequent collapse of alveolar structures and massive epithelial cell death while the other glands of the same animal remain morphologically and functionally in a lactating state. Many markers of involution such as sulfated glycoprotein-2, protein kinase A, transcription factor AP-1 and most notably stromelysin are induced in sealed glands. These findings suggest a cell death pathway which is independent of the systemic levels of lactogenic hormones but which is triggered by an accumulation of apoptosis-inducing factors in the milk, in the lobulo-alveolar structures or by a physical distortion of secretory epithelial cells generated by the engorgement. PMID- 9208230 TI - Is the sensitivity of cells for FGF-1 and FGF-2 regulated by cell surface heparan sulfate proteoglycans? AB - We have examined the importance of cell surface heparan sulfate proteoglycans (HSPG) in fibroblast growth factor (FGF) signaling. 3T3 cells grown under conventional conditions were much more sensitive to FGF-2 compared to FGF-1. However, cells were equally sensitive to FGF-1 and FGF-2 using conditions which reduced the effect of endogenous HSPG. Addition of heparin, or treatment with chlorate, an inhibitor of proteoglycan sulfation, resulted in enhanced or reduced growth factor response, respectively, and eliminated the differences between FGF 1 and FGF-2. HSPGs isolated from trypsin digests of 3T3 cells had a much higher affinity for FGF-2 compared to that for FGF-1 when analyzed by affinity chromatography. Glycosaminoglycan chains or core protein fragments derived from the HSPG failed to show the same high apparent affinity for FGF-2, suggesting that an intact proteoglycan structure was important for the high FGF-2 affinity. Addition of HSPG ectodomains, isolated from cultured 3T3 cells or produced as recombinant molecules, to chlorate-treated cultures of 3T3 cells inhibited the mitogenic activity of FGF-2 and eliminated the effect of heparin as a potentiator of either growth factor. These results support the idea that the cell-associated HSPG is an integral component of the FGF signaling system and in 3T3 cells contributes to the increased sensitivity of these cells to FGF-2 compared to FGF 1. Since isolated ectodomains of HSPG inhibited rather than stimulated the mitogenic response of the FGFs, the proper anchoring of the HSPG in the cell membrane appears to be important for a stimulatory effect. PMID- 9208231 TI - Characterization of polypeptides in Euglena gracilis which are synthesized in a circadian manner. AB - In vivo labeling of proteins in the unicellular phytoflagellate Euglena gracilis, grown under constant light, with [35S]methionine, gave evidence that three proteins of 17,000, 24,000 and 60,000 M(r) were synthesized rhythmically with a period length of 26 h. The 60,000 M(r) protein was synthesized at the subjective end of the 12 h light phase, while the two smaller proteins of 17,000 and 24,000 M(r) shared a maximum of synthesis at the subjective noon. The rate of synthesis oscillated up to 20-fold when the maxima and minima were compared. By isoelectric focusing, the 24,000 M(r) protein was separated into three distinct spots. When different temperatures between 16 and 27 degrees C were applied, the period length of the circadian synthesis rhythms differed by only 3 to 5 h, indicating that the synthesis rhythms of these proteins were almost temperature compensated. Synthesis of the two smaller peptides was inhibited by cycloheximide but not by chloramphenicol, implying that synthesis occurred at 80S ribosomes. Once synthesized and processed, these two proteins were fairly stable in continuous light for about 120 h, while they were degraded during darkness- although slowly. After cell breakage, these proteins were localized in the pelletable membranous fraction. PMID- 9208232 TI - ADP-ribosylation of Rho-proteins with botulinum C3 exoenzyme inhibits invasion and shape changes of T-lymphoma cells. AB - C3 exoenzyme from Clostridium botulinum ADP-ribosylates the small GTP-binding protein Rho with a high specificity. The use of C3 has shown that Rho-mediated signaling is involved in the regulation of actin-dependent processes in various cell types. In order to investigate the role of Rho-proteins in lymphocyte crawling, we have analyzed the effects of C3 on a T-cell line derived from the murine BW5147 lymphoma. Pretreatment of the lymphoma cells with C3, in conditions where Rho was actually ADP-ribosylated, strongly inhibited the characteristic shape changes resulting from extension and retraction of pseudopodia. Concomitantly, invasion of the cells through a monolayer of fibroblast-like cells was also inhibited. C3-treatment did not affect the total F-actin content of the cells, as measured by flow cytometry of cells stained with phalloidin. Yet, microscopical observation revealed that the accumulations of F-actin, which were seen in the pseudopodia of untreated cells, were absent after treatment with C3. This suggests that C3 may affect actin polymerization locally. The inhibitory effect of C3 on invasion was not restricted to the murine BW5147 lymphoma cell line, as it occurred also with CCRF-CEM, a human T-cell lymphoma line. Our results demonstrate that invasion-bound motility of lymphocytes depends on a Rho mediated signal transduction pathway. PMID- 9208233 TI - A review of teicoplanin in the treatment of serious neonatal infections. AB - Gram-positive bacteria, notably coagulase negative staphylococci, have become an important cause of infection in neonates. Furthermore, many of these pathogens are now resistant to multiple antibacterial agents. Teicoplanin, a glycopeptide antibiotic, is active against a broad range of Gram-positive pathogens, including methicillin-resistant staphylococci. It has advantages over vancomycin in terms of tolerability, with a lower propensity to cause nephrotoxicity and anaphylactoid-like reactions, and in terms of ease of administration and monitoring requirements. The clinical utility of teicoplanin in neonates with Gram-positive infections has been investigated in several noncomparative studies. Clinical and bacteriological response rates in 173 neonates treated with teicoplanin 8-10 mg/kg intravenously or intramuscularly once daily after a loading-dose regimen of 10-20 mg/kg per day have ranged from 80%-100% and 83% 100%, respectively. Few adverse events related to teicoplanin have been reported in this patient population. CONCLUSION: Teicoplanin (8-10 mg/kg) administered intravenously or intramuscularly once daily after a loading-dose regimen of 15-20 mg/kg per day appears to be an effective and well tolerated treatment for Gram positive infections in neonates. PMID- 9208234 TI - Use of albumin in neonatal resuscitation. AB - The use of albumin plasma has become popular during resuscitation of the term baby with very low Apgar scores (< or = 2 at 1 min). There is no evidence of benefit from this practice which may actually be damaging to babies with severe asphyxia causing myocardial damage. PMID- 9208235 TI - Generalized metaphyseal modification with cone-shaped epiphyses following long term administration of 13-cis-retinoic acid. AB - We report on a 6-year-old girl with short stature which developed following the administration of 13-cis-retinoic acid (a synthetic derivative of vitamin A or retinoid) for 40 months as adjunct chemotherapy for neuroblastoma. Radiographic examination suggested osteophyte formation in the cervical spine, which is the most common skeletal manifestation of retinoid toxicity [10, 11]. In addition, severe metaphyseal cupping with a cone-shaped epiphysis primarily affecting rapidly growing long bones was found, which represented impaired enchondral ossification. This epi-metaphyseal alteration, though unusually severe, was reminiscent of the premature epiphyseal closure which has been described as an adverse effect of 13-cis-retinoic acid [10-12]. Other minor skeletal changes included posterior scalloping of the vertebral bodies and increased interpediculate distances, which were related to a widened spinal canal found on CT. A literature search disclosed several primary skeletal dysplasias with superficial radiological similarities to those of the present patient. However, these entities showed significant clinical and radiological differences from our patient. CONCLUSION: The precise cause of the generalized skeletal alteration in the present patient remained unknown, but it conceivably resulted from the administration of 13-cis-retinoic acid. PMID- 9208236 TI - Seasonal variation in childhood asthma hospitalisations in Finland, 1972-1992. AB - All hospital treatment periods caused by asthma in children under 15 years in Finland during 1972-1992 were examined. The data were obtained from the Hospital Discharge Register, covering all hospitalisations in Finland. A total of 59,624 asthma related treatment periods were recorded. The monthly variation in hospitalisations peaked in May (35.6% above the trend) and in autumn and early winter (41.3% above the trend in October), whereas the monthly variations were low in late winter and in summer. The overall profile of seasonal variation was similar in both sexes, although admissions were lower for boys than for girls in winter and higher in autumn. The average monthly deviation was highest in the age group 0-4 years in May, 42.8% above the trend, and highest in the age group 5-9 years in October, 53.9% above the trend. Closer examination of the seasonal variation gives indirect information on possible trigger factors for acute asthma. CONCLUSION: A clear seasonal variation could be observed in childhood asthma hospital admissions, together with age and sex-related differences in this seasonality. Preventive treatment for asthma should be used effectively in order to avoid acute attacks leading to hospitalisation in children who are allergic to birch pollen and also at times of viral respiratory infections. PMID- 9208237 TI - Herpes virus (KSHV) associated Kaposi sarcoma in a 3-year-old child with non-HIV induced immunodeficiency. AB - A 3-year-old boy of German descent suffered from two episodes of Streptococcus pneumoniae meningitis within 2 months. One month previously, the first skin lesion of Kaposi sarcoma (KS) had been observed behind his right ear. During the following 2 years KS disseminated not only mucocutaneously but also to visceral organs. Immunological evaluation revealed severe lymphocytopenia with reduced helper/suppressor T-cell ratio and impaired humoral immune response to pneumococci. Extensive laboratory tests gave no evidence for known immunocompromising infections. However, recently described DNA sequences from a Kaposi sarcoma-associated herpesvirus (KSHV) could be identified within skin tissue. As chemotherapy failed to stop tumour progression the patient was referred for bone marrow transplantation. Eighteen months later the KS is in remission and the patient in a good general condition. CONCLUSION: The case supports the hypothesis that KSHV is involved in the aetiology of KS. Bone marrow transplantation is possibly a therapeutic option for KS in patients with immunodeficiency not related to human immunodeficiency virus infection. PMID- 9208238 TI - Depressed natural killer cell activity due to decreased natural killer cell population in a vitamin E-deficient patient with Shwachman syndrome: reversible natural killer cell abnormality by alpha-tocopherol supplementation. AB - Natural Killer (NK) cell activity was examined in a 16-month-old Japanese boy with Shwachman syndrome associated with severe vitamin E deficiency. As evaluated by 51Cr-release assay from K562 cells, NK cell activity was constantly decreased. After 8 weeks of oral alpha-tocopherol (alpha-Toc) supplementation (100 mg/day), NK cell activity had normalised. When alpha-Toc supplementation was interrupted for 16 weeks. NK cell activity again decreased. Flow cytometry of peripheral lymphocytes revealed a lowered number of CD16+ CD 56- fraction, which has the most potent NK cell activity. Single cell-in-agarose assay, to investigate the binding and cytolytic activity of NK cell at the single cell level, revealed that the number of NK cells which bind to K562 cell was decreased, but that the cytolytic activity of the individual binding cell was relatively unaffected. A second supplementation of alpha-Toc for 8 weeks successfully restored NK cell activity, the number of cells expressing NK cell markers and the number of K562 binding cells as compared to the age-matched normal range. CONCLUSION: These results indicate that severe vitamin E deficiency caused impaired NK cell activity due to a decrease in the number of CD16+ CD56- NK cells and that this abnormality is reversible with alpha-Toc supplementation. PMID- 9208239 TI - Natural killer cell activity and hypovitaminosis E. PMID- 9208240 TI - Plasma fibronectin levels in meningococcal disease. AB - Fibronectin (a glycoprotein which modulates inflammation) may decrease mortality in systemic infection. Children with meningococcal disease (MCD) may have low fibronectin levels. We aimed to compare plasma fibronectin levels in children with MCD and controls, correlate fibronectin levels with interleukin-6 (IL-6), shock and death, and assess fibronectin as an aid to early diagnosis in MCD. Samples were taken on admission from 99 children with MCD and 49 controls. Plasma fibronectin was measured using a turbidimetric immunoassay. Plasma fibronectin was significantly lower in MCD compared to controls (57 micrograms/ml vs 105 micrograms/ml; P < 0.005). Children who died had significantly lower levels than survivors (29 micrograms/ml vs 62 micrograms/ml; P = 0.01). Fibronectin levels were negatively correlated with IL-6 levels. Fibronectin was a poor predictor of MCD. CONCLUSION: Plasma fibronectin levels are decreased in children with MCD, especially in shock and death. This decrease is associated with high IL-6 levels. Fibronectin could be a novel therapy in severe MCD. PMID- 9208241 TI - Hepatitis B and C virus infection in Polish children with malignancies. AB - Infections by hepatotropic viruses belong to the most common complications of chemotherapy in children suffering from neoplastic diseases. The rate of hepatitis B virus (HBV) or hepatitis C virus (HCV) infection and the effectiveness of passive immunization against HBV were studied in 285 children; 148/285 with lymphoproliferative diseases and 137/285 with solid tumours. HBV infection was observed in 10.2% children receiving hepatitis B immune globulin as compared to 36.8% without passive immunization against HBV. Anti-HCV antibodies were similar in both groups amounting 38.7% and 32.6% respectively. CONCLUSION: The results show that hepatitis B immune globulin administration is effective and that HCV might become the main cause of hepatitis among immunosuppressed patients in the future. PMID- 9208242 TI - The clinical significance of rigors in febrile children. AB - The objective of the study was to evaluate the significance of rigor as a predictor of bacterial infection in hospitalized febrile infants and children. One hundred febrile children with rigor were studied and compared to 334 febrile matched controls without rigor. All underwent clinical evaluation and appropriate laboratory investigations. The patients were then divided into "bacterial" and "non bacterial" infection groups, as defined in the text. It was demonstrated that 66% of the patients with rigor belonged to the bacterial infection group versus 50% in the non-rigor group (P < 0.005). There was a significantly greater yield of positive blood cultures in the patients with rigor (P < 0.04), especially those over the age of 1 year (P < 0.015). The only laboratory examination of potential value as a predictor of bacterial infection in children with rigor was the band count. An absolute band count of more than 1500/mm was significantly more frequent in the rigor group (P < 0.003), and the combination of a rigor and band count of more than 1500 increased the relative risk for a bacterial infection by a factor of 1.35. These data demonstrate that rigor in hospitalized febrile infants or children significantly increase the likelihood of bacterial infection. CONCLUSION: Although the absence of rigors in febrile children does not exclude bacterial aetiology, their presence significantly increase the probability of an infection requiring appropriate workup and a reader institution of antibiotic therapy. PMID- 9208243 TI - Technical considerations for inhaled nitric oxide therapy: time response to nitric oxide dosing changes and formation of nitrogen dioxide. AB - The aim of the present study was to analyse the time response to nitric oxide (NO) dosing changes as well as the formation of nitrogen dioxide (NO2) with different ventilation systems, respirator settings and application sites during NO inhalation. The inspired NO and NO2 concentrations were continuously measured using chemiluminiscence within a dummy ventilatory system equipped with two different respirator systems (Siemens Servo 900c and Bear BP 2001). NO was either introduced into the afferent limb of the ventilatory circuit close to the endotracheal tube (site A) or into the so-called low pressure port of the Servo 900c respirator, far away from the endotracheal tube (site B). In addition, the decay of the inspired NO concentration after cessation of the NO gas flow was studied. This decay was considerably prolonged when NO was introduced at site B (time constants: tau = 7.19 min versus tau = 0.29 min). Within the concentration range studied (0-25 ppm NO) a linear correlation between the NO and NO2 concentration was found. At site A and an inspired oxygen concentration of > 0.95 NO2 formation amounts to 1.14% +/- 0.11% of the NO concentration. Using this value one can calculate the NO2 formation for a given NO dose. For example, when 40 ppm NO are applied, a concentration of 0.45 ppm NO2 can be expected, which is well below the relevant toxic concentrations. However, when NO was introduced at site B, NO2 formation was significantly increased to 1.61% +/- 0.16%. Passage of the ventilated gas through soda lime led only to a slight and insignificant reduction in NO2 concentration. The continuous flow respirator BP 2001 showed a significantly lower NO2 concentration when compared to the non-continuous flow respirator Servo 900c (0.64 +/- 0.11% vs.1.14 +/- 0.11%). CONCLUSION: The application of NO close to the endotracheal tube is associated with a much faster response of the actual inspired NO concentration to dosing changes and shows the lowest NO2 formation. In order to avoid toxic NO2 concentrations, an upper limit of 40 ppm NO is recommended for continuous NO inhalation. PMID- 9208244 TI - The Marshall-Smith syndrome: a review of the laryngeal complications. AB - The Marshall-Smith syndrome is characterised by a triad of facial dysmorphism, failure to thrive and accelerated osseous maturation. We report a further case of this rare syndrome with the unusual but previously reported complication of laryngeal hypoplasia and review the associated laryngeal anomalies that have been reported to date. CONCLUSION: Severe airway obstruction due to congenital anomalies must be excluded in any dysmorphic child presenting with respiratory distress at birth. Rapid airway assessment will enable early and appropriate intervention and may be important when deciding on the long-term plan for the infant. PMID- 9208245 TI - Blood transfusion. Iron load and retinopathy of prematurity. AB - To study the relationship between blood transfusion, iron load and retinopathy of prematurity (ROP), we performed a prospective observational cohort study in a level III neonatal intensive care unit. During a 24-month period, data on the volume of blood transfused during the first 6 weeks of life and on the incidence of ROP were collected in all surviving very low birth weight infants (n = 114; median birth weight 1130 g. range 520-1500 g). Associations between these data and values for serum iron, transferrin and ferritin measured at weekly intervals were analysed in a nested case-control design by logistic regression. There was a significant association between the volume of blood transfused and the incidence of ROP. After adjustment for gestational age at birth, duration of oxygen therapy (FiO2 > 0.3) and duration of mechanical ventilation, the relative risk of developing ROP was 6.4 (95% CI 1.2-33.4) for infants who had received 16-45 ml/kg, and 12.3 (1.6-92.5) for those who had received more than 45 ml/kg of blood (reference, 0-15 ml/kg). In contrast, there was no independent relationship between ROP and any of the parameters on iron metabolism analysed. CONCLUSION: This study confirms the role of blood transfusions as an independent risk factor for ROP. This relationship, however, does not appear to be mediated via an increased iron load. PMID- 9208246 TI - Conjunctival impression cytology in the preterm infant and it's relation to outcome. AB - The preterm infant is deficient in vitamin A (retinol) and this has been implicated in the pathogenesis of chronic lung disease of prematurity. Conjunctival impression cytology (CIC) has been used in adults to assess retinol status. We aimed to assess the feasibility of performing CIC in the preterm infant and to determine the significance of abnormal CIC findings. CIC samples were collected during routine retinopathy screening, and classified as inadequate, normal, borderline normal or abnormal. Ninety preterm infants were studied. Seventy-four (82%) CIC specimens produced a positive yield, whereas 16 (18%) were inadequate. Of the 74 adequate samples, 61 (82%) were normal or borderline normal and 13 (18%) abnormal. Seventy-three CIC specimens were assessed by a second histopathologist with complete agreement on 64 (88%) samples and disagreement on 9 (12%) samples. Ten sets of conjunctival impressions, taken from both eyes, gave identical results in all adequate samples. Birth weight was significantly lower in this abnormal group. Four infants (32%) in the abnormal group required treatment for retinopathy compared to two (3%) in the normal/borderline normal group, (P < 0.01). CONCLUSION: Conjunctival impression cytology is simple and reproducible technique which maybe easily applied to the preterm infant. Abnormal CIC is associated with retinopathy of prematurity requiring treatment. PMID- 9208247 TI - Seizure risk in offspring of individuals with a history of febrile convulsions. AB - One-hundred and seventy-nine offspring of 120 probands with a history of febrile convulsions (FC) were studied to determine the risk of seizures and possible factors influencing this risk. The conditions for this study were especially good since all of the probands had undergone clinical and EEG examinations as well as an assessment of family history of seizures during childhood. Hence, for the first time the seizure status of the probands' parents could be included in the calculation of risk in offspring. In sibs the risk was highest if the mother of the proband had experienced seizures (20% vs 9% in offspring of probands with nonaffected parents). Similarly, offspring of probands with affected mothers had a much higher risk (27%) than offspring of probands with affected fathers (7%). Our findings point to a maternal preponderance in the transmission of FC liability. No relationship was found between the presence of EEG traits of a genetic seizure liability (theta rhythms, spikes and waves, photoparoxysmal response, focal sharp waves) in probands during childhood and the seizure risk in their offspring. The present data provide no basis for forming an hypothesis regarding the possible mode of inheritance of FC. This is not surprising since FC as already shown in the EEG-are not a homogeneous disorder, but are caused by a variety of genetic factors occurring in variable constellations. Possibly, in a subgroup of probands with seizure affected mothers the susceptibility to FC follows a multifactorial polygenic mode of inheritance. CONCLUSION: The seizure incidence in offspring of individuals with a history of FC was 10% (only FC in 64% of the affected offspring). Offspring of females with affected parents were at an increased risk. Pathological childhood EEG findings of the probands were not related to an increased risk in offspring. PMID- 9208248 TI - Diagnostic delay in neurofibromatosis type 1. AB - Since 1985 a multidisciplinary team in the Sophia Children's University Hospital in Rotterdam provides diagnostic follow up and genetic counseling services for neurofibromatosis type 1 (NF1) patients and their families. Parents of 68 affected children as well as 24 affected parents were interviewed. Of the affected children, 50% and 33% of the affected adults were treated for symptoms related to NF1 before a specific diagnosis was made. Although the disease is fully penetrant by the age of 5 years, 35% of the affected children had not been diagnosed by this age. Parents stated a preference for early diagnosis of NF1. Diagnosis of NF1 did not seem to be a reason to refrain from having children. The general attitude towards prenatal diagnosis was positive; however few parents would actually terminate an affected pregnancy. CONCLUSION: Overall delay in diagnosis of NF1 is significant. Knowledge of symptoms should make an early diagnosis possible with beneficial effects for the patient and family members. PMID- 9208249 TI - Ascorbic acid enhances hydroxyl radical formation in iron-fortified infant cereals and infant formulas. AB - Infant cereals and formulas are usually fortified with iron to prevent iron deficiency. To enhance iron bioavailability, supplemental ascorbic acid is recommended. Ascorbic acid is considered to be an antioxidant in vivo, but has pro-oxidant effects when exposed to non-protein-bound iron. We measured formation of free radicals in cereals and infant formulas after addition of ascorbic acid. The production of hydroxyl radicals was assessed by hydroxylation of salicylic acid to 2.5-dihydroxybenzoic acid (2,5-DHBA). Production of 2.5-DHBA increased with increasing ascorbic acid doses added. Addition of 0.8 mM ascorbic acid to breast milk produced less radicals (0.03 +/- 0.05 microM) than addition of ascorbic acid to low-iron formula (0.13 +/- 0.08 microM. P = 0.019), medium-iron formula (0.34 +/- 0.12 microM, P < 0.0001) or high-iron formula (0.44 +/- 0.08 microM. P < 0.0001). Even when iron content in breast milk was adjusted to a level comparable with that of formulas, production of 2,5-DHBA was lower. Breast milk seems to contain substances that reduce hydroxyl radical formation. CONCLUSION: Supplemental ascorbic acid causes hydroxyl radical formation in iron fortified infant nutrients in vitro. PMID- 9208251 TI - Multiple chemical sensitivity. A view from under the grassroots. PMID- 9208252 TI - Case of the month: a 10-year-old boy with Down syndrome and ichthyosiform changes. PMID- 9208250 TI - Inhibition of enteropathogenic Escherichia coli adhesion to HEp-2 cells by colostrum and milk from mothers delivering low-birth-weight neonates. AB - Breast milk samples from three groups of Brazilian women were evaluated for their inhibitory effect on enteropathogenic Escherichia coli (EPEC) adhesion to HEp-2 cells: G1, mothers delivering preterm babies of appropriate birth weight (n = 12); G2, mothers delivering term babies of low birth weight (n = 11); G3, the control group, mothers delivering term babies of appropriate birth weight (n = 39). Colostrum samples were obtained at 48-72 h and milk samples on the 7th, 30th and 60th days after delivery. All samples showed strong inhibitory activity (66% 100%), without significant differences among the three groups and four periods. Total IgA and anti-EPEC IgA concentrations were significantly higher in colostrum than in milk samples in the three groups studied. The levels of colostral IgA and anti-EPEC IgA observed in G1 and G2 were significantly higher compared to the control group. Western blotting assays showed that individual samples as well as pools of colostrum or milk samples contain IgA antibodies to many EPEC outer membrane proteins. A 94 kDa band with molecular weight consistent with the EPEC adhesin named intimin; was recognized by all samples analysed. Bands of different molecular weight were also recognized by some samples of colostrum and milk, such as a band of approximately 18.4 kDa, with molecular weight equivalent to bundle forming pilus subunits. CONCLUSION: Our results suggest that colostrum and milk from mothers of premature and small-for-date term neonates are as effective in protecting the newborn against EPEC infections as those from mothers of term babies of appropriate birth weight. PMID- 9208253 TI - Familial Mediterranean fever and polyarteritis nodosa: experience of five paediatric cases. A causal relationship or coincidence? PMID- 9208254 TI - Acute pancreatitis associated with food allergy. PMID- 9208255 TI - Elevated interleukin 6 without pleocytosis in cerebrospinal fluid in encephalitis patients. PMID- 9208256 TI - Exogenous surfactant therapy for status asthmaticus. PMID- 9208257 TI - How to prevent stroke recurrence in patients with patent foramen ovale: anticoagulants, antiaggregants, foramen closure, or nothing? AB - Several studies found a significant association between patent foramen ovale (PFO), interatrial septal aneurysm, and patients less than 60 years of age presenting with acute stroke and without any identified coexisting mechanism explaining the acute event. Paradoxical embolism from a venous source through a right-to-left shunt is usually incriminated, but the definite proof for paradoxical embolism is often lacking, with screening for deep-venous thrombosis leading to variable estimates. Despite these controversies, the-possibility of paradoxical embolism in patients with cryptogenic stroke and PFO is commonly retained as the cause of the neurological deficit. Moreover, there are now definite studies documenting that these patients are at risk of recurrence. The aim of the present paper is to review the literature on the risks of stroke recurrence in patients with atrial septal defects, and to weigh the risks and benefits of the different therapeutic options currently available to prevent stroke recurrence. These options include chronic oral anticoagulant or antiplatelet therapy, and more invasive procedures such as surgical closure or transcatheter closure of the defect. Finally, using the principles of decision analysis, the authors suggest tentative practical therapeutic recommendations that might be helpful to clinicians in daily practice. PMID- 9208258 TI - Treatment of patent foramen ovale and stroke: to close or not to close, that is not yet the question. PMID- 9208259 TI - Cerebrovascular disease as the initial clinical presentation of haematological disorders. AB - We describe 14 patients (mean age 57 years) in whom stroke or TIA was the presenting manifestation of a haematologic disorder. Twelve patients had an ischaemic stroke and 2 a haemorrhagic stroke. This group represented 1.27% (14/1,099) of the total number of patients with first-ever stroke diagnosed from 1986 to 1992 at our institution, accounted for 1.32% (12/906) of all brain infarcts and 1.03% (2/193) of all haemorrhagic strokes, and was the most common aetiology (25%) of ischaemic stroke of unusual cause. Haematological disorders included essential thrombocythaemia (6), polycythaemia vera (1), smoker's polycythaemia (1), thrombotic thrombocytopenic purpura (1), IgA lambda myeloma (1), acute lymphoblastic leukaemia (1), Waldenstrom's macroglobulinaemia (1), chronic granulocytic leukaemia (1) and IgG lambda myeloma (1). Stroke subtypes included definitive cerebral infarct (10), TIA (2), parenchymal haemorrhage (1) and spontaneous subdural haematoma (1). Vascular territories in ischaemic stroke were the carotid in 7 patients, the vertebrobasilar in 1 and undetermined in 4. Mean follow-up was 40 months (range, 1-96 months). The mortality rate was 18.7%. PMID- 9208260 TI - Coenzyme Q10 treatment in mitochondrial encephalomyopathies. Short-term double blind, crossover study. AB - We report a short-term double-blind, crossover study of CoQ10 in 8 patients with mitochondrial encephalomyopathies. Four patients had myoclonus epilepsy with ragged-red fibers syndrome, 3 had mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes syndrome, and 1 had chronic progressive external ophthalmoplegia with myopathy. A trend of effectiveness of CoQ10 in several parameters was noted. Fatigability of daily activities was alleviated. The endurance to muscle exercise was augmented. Global muscle strength scored by Medical Research Council scale was increased. The extent of elevation in serum lactate and pyruvate levels after exercise was decreased. However, only the global MRC index score had a statistical significance (p < 0.05). There were no side effects during therapy. The serum CoQ10 levels were significantly lower in patients than in normal controls before CoQ10 treatment and increased significantly after treatment. PMID- 9208261 TI - Quantitative EEG and statistical mapping of wakefulness and REM sleep in the evaluation of mild to moderate Alzheimer's disease. AB - Statistical probability mapping was used to quantify and localize EEG differences between 27 patients with Alzheimer's disease (AD) and 25 age- and gener-matched controls. Differences in mean activity in four EEG frequency bands (delta, theta, alpha, beta) for wakefulness and for REM sleep were examined, t-statistic maps clearly highlighted common pattern anomalies in AD patients in the two states. More specifically, Alzheimer patients were more affected than control subjects in parieto-temporal and frontal regions. These differences were more prominent in REM sleep and consisted primarily in an increase in absolute delta and theta activities, and a decrease in absolute alpha and beta activities. Discriminant analysis, using a ratio of slow over fast frequencies, yielded a classification rate of 90.4% (sensitivity 81.5%, specificity 100%) for REM sleep. For wakefulness, the same measure allowed correct classification of 80.8% of the subjects (sensitivity 66.7%, specificity 96%). PMID- 9208262 TI - Hypovitaminosis D and decreased bone mineral density in amyotrophic lateral sclerosis. AB - To assess the bone health of patients with amyotrophic lateral sclerosis (ALS), we evaluated the bone density and serum biochemical indices of bone metabolism in 11 ALS patients. The serum concentration of 25-hydroxyvitamin D (25-OHD) was significantly lower in patients (14.0 +/- 3.7 ng/ml) than in controls (25.2 +/- 4.0 ng/ml), at deficient levels (< 10 ng/ml) in 2, and at insufficient levels (10 20 ng/ml) in 9 patients. Serum levels of parathyroid hormone (PTH) and ionized calcium were elevated in 8 and 6 patients, respectively. Dietary intake of vitamin D was below the recommended level (100 IU) in 10 patients, and 10 patients were in a sunlight-deprived state. The metacarpal bone density (MBD) and the metacarpal index (MCI) of the second metacarpal bone were measured by computed X-ray densitometry. Z scores of the MBD and the MCI were negative in 7 and 6 patients, respectively. The serum concentration of 25-OHD was positively correlated with the Z score of the MBD (p < 0.05, r = 0.727) and negatively with the PTH level (p < 0.05, r = -0.410). The degree of dysfunction of hand grip also correlated with the Z score of the MBD (p < 0.05, r = 0.749). These data underscore the importance of hypovitaminosis D and compensatory hyperparathyroidism in the development of osteopenia in patients with ALS. PMID- 9208263 TI - Persisting childish behavior after bilateral thalamic infarcts. AB - We report a case with bilateral paramedian thalamic infarcts. The patient showed a dramatic personality change characterized by childish behavior and euphoria; which remained unchanged for 2 years after the onset. 'Vorbeireden' characterized by approximate answers was also observed. Anterograde amnesia had quite improved after 2 years, while retrograde amnesia for 1 year prior to the stroke onset and vertical gaze palsy remained unchanged. An MRI scan demonstrated bilateral medial thalamic and right midbrain infarcts without other lesions in the brain. A position emission tomography study showed that cerebral metabolic rate for glucose was markedly decreased in both thalami and in the cerebellum, and only slightly decreased in the parietal and occipital cortical regions. Cerebral metabolic rates of glucose in the frontal and temporal cortices were within normal range. The paramedian thalamic lesions per se may be responsible for the patient's personality change, 'Vorbeireden', and amnesia. PMID- 9208264 TI - Improvement of motor fluctuations in patients with Parkinson's disease following treatment with high doses of pergolide and cessation of levodopa. AB - The combination of levodopa and a dopamine agonist in the treatment of patients with Parkinson's disease often reduces the severity of motor fluctuations. In patients with very severe motor fluctuations, monotherapy with continuous subcutaneous infusions of the dopamine agonist apomorphine may result in a marked reduction of hyperkinesia and on-off phenomena. We report 3 patients with Parkinson's disease and motor fluctuations who received high doses of pergolide without levodopa resulting in a reduction of motor fluctuations. All patients received doses of pergolide exceeding the maximum recommended dose. One patient also required additional therapy with amantadine. These data show that in some patients oral treatment with high doses of a dopamine agonist may improve the severity of motor fluctuations and achieve a good control of parkinsonian signs without concomitant levodopa treatment. PMID- 9208265 TI - A comparative study of isokinetic dynamometry and manual muscle testing of ankle dorsal and plantar flexors and knee extensors and flexors. AB - Muscle strength in neuropathic patients is usually evaluated clinically using manual muscle testing (MMT). Detection and grading of mild symmetrical muscle weakness using MMT is difficult partly because the examiner must take into consideration the normal variation in strength in relation to age, weight, height, and gender. In the present study assessment of the strength of ankle dorsal and plantar flexors and knee flexors and extensors with MMT and isokinetic dynamometry were compared in 108 patients, of whom 86 had diabetes mellitus and 22 had alcoholic liver cirrhosis. The isokinetic muscle strength of the patients was compared with the strength of 90 healthy control subjects, adjusted for the influence of age, weight, and height for both genders. MMT resulted in a significant underestimation of the frequency and severity of muscle weakness in both the ankle and the knee. In 28-41% of the comparisons, MMT misclassified the strength performance with one category or more (> 25%). Misclassifications were most frequent for the ankle plantar flexors. PMID- 9208266 TI - Cerebrospinal-fluid ciliary neurotrophic factor in neurological patients. AB - We developed a double sandwich immunoassay for the dosage of ciliary neurotrophic factor (CNTF) in cerebrospinal fluid (CSF). The detection limit was 100 pg/ml. This assay was applied to human CSF samples from 14 normal subjects, 26 patients with multiple sclerosis (MS), 17 with Guillain-Barre syndrome (GBS) or chronic inflammatory demyelinating polyneuropathy (CIDP), and 22 with tumours of the central nervous system (CNS) or leucaemic meningosis (LM). Samples from normal control subjects and from patients with tumours did not contain detectable CNTF. Only 2 patients with LM were positive, and all the patients with inflammatory diseases of the CNS and peripheral nervous system were positive. The MS group presented a mean value of 240 pg/ml CNTF and the GBS/CIDP group a value of 430 pg/ml. PMID- 9208267 TI - A case of acute transverse myelitis affecting the entire length of the spinal cord. PMID- 9208268 TI - Autonomous stump movements in brain death. PMID- 9208269 TI - MR monitoring of a medically treated spinal cord abscess presumptively due to Listeria monocytogenes. PMID- 9208270 TI - Increased cerebral glycogen detected by localized 1H-magnetic resonance spectroscopy in a patient with suspected McArdle's disease. PMID- 9208271 TI - Cheiro-oral syndrome and vivid recollection of past in thalamic infarction. PMID- 9208272 TI - Atypical white matter changes in Alzheimer's disease. PMID- 9208273 TI - Regression of a conservatively treated gigantic fusiform aneurysm of the middle cerebral artery. PMID- 9208274 TI - Hereditary neuropathy with liability to pressure palsies (HNPP) revealed after weight loss. PMID- 9208275 TI - Experience with antisense oligodeoxynucleotides in renal cells. PMID- 9208276 TI - Heparan sulphate proteoglycans and diabetic nephropathy. PMID- 9208277 TI - Phospholipase A2 in mesangial cells: control mechanisms and functional importance. PMID- 9208278 TI - Potential uremic toxins modulate energy metabolism of cardiac myocytes in vitro. AB - In the present study we investigated the direct effects of potential uremic toxins on the energy metabolism of cultured cardiac myocytes. High-energy phosphates were extracted with perchloric acid and determined by high performance liquid chromatography. Energy charge (calculated from the ratio of [ATP], [ADP] and [AMP] was significantly reduced by 20 mM urea and the combination of creatinine (5 mM) plus urea (200 mM). On the other hand, perfusion with culture media containing clinically relevant amounts of urea (20 mM) or creatinine (1 mM) increased the PCr/ATP ratio. This effect was more pronounced after application of an artificial uremic medium (consisting of uremic serum, urea, creatinine and cytokines) or high amounts of creatinine (5 mM) plus urea (200 mM). As contractility of myocytes is reduced due to application of uremic compounds or uremic serum, we attribute changes in contraction frequency or inotropy to dysregulation of calcium availability within the cell. In fact, the cardiodepressive action of uremic serum (2.5%) could be completely reversed by the calcium agonist, Bay K 8644, thus indicating disturbances in myocardial calcium homeostasis in uremia. Altered calcium regulation by uremic toxins might therefore be responsible for the observed changes in myocardial energy metabolism. These results might contribute to the understanding of the pathogenesis of cardiac damage in end-stage renal disease. PMID- 9208279 TI - Glomerular number and size following chronic angiotensin II blockade in the postnatal rat. AB - Chronic angiotensin-converting enzyme (ACE) inhibition with enalapril or angiotensin II (Ang II) receptor antagonism with either losartan (specific for Ang II type-1 receptor, AT1) or PD 123319 (specific for the Ang II type-2 receptor, AT2) were effected between postnatal days 3 and 21 in the rat. Following quantitative analysis of the kidneys using recently developed unbiased stereological techniques we found that none of the treatments resulted in changes in glomerular number or size. This implies that inhibition of Ang II activity had no effect on postnatal nephron induction or glomerular development. However, following both chronic ACE inhibition and AT1 antagonism, abnormalities of tubules and their associated vessels were evident throughout the kidney and were accompanied by an increased proportion of interstitium. The structural abnormalities were most prominent in the outer medulla and were consistent with interruption of descent of the loops of Henle and vasa rectae. In contrast, no renal morphological abnormalities were observed following chronic AT2 antagonism. PMID- 9208280 TI - Myoglobinuric acute renal failure in the rat: a role for acidosis? AB - Myoglobin induces renal injury by mechanisms that remain incompletely defined. Acidosis has been suggested as an important factor in myoglobinuric renal failure, and urine alkalization is routinely recommended for its prevention. We tested this hypothesis by exploring the effects of acid-base balance upon myoglobin nephrotoxicity in vivo and in vitro. In isolated rat kidneys at normal pH, myoglobin at concentrations of 25-250 mg/dl minimally affected renal perfusion flow, glomerular filtration rate (GFR) and tubular sodium reabsorption (TRNa). By contrast, at pH 7.1 myoglobin induced vasoconstriction, reduced GFR and TRNa and increased hypoxic injury to medullary thick ascending limbs. These changes were largely reproduced by perfusing kidneys with hematin, suggesting its release from myoglobin in acidosis. Chronic alkalosis or acidosis was induced in rats by supplementing drinking water with 0.28 M NaHCO3 or NH4Cl, respectively. Acute renal failure, produced in control animals by myoglobin infusion (38 mg/100 g body weight), was comparably prevented by both chronic alkalosis and acidosis. Acute intravenous or oral acid load provided similar protection. Thus, although acidosis exacerbates myoglobin toxicity in isolated perfused kidneys, acute or chronic exogenous acid load prevents renal damage in vivo. This may underscore the protective properties of solute load, a consequence of preconditioning, and suggests that, in the crush syndrome, endogenous acidosis rather than being an independent risk factor is a marker of tissue hypoperfusion and organism susceptibility to myoglobin renal toxicity. PMID- 9208281 TI - Effect of the antiglucocorticoid RU-486 on glomerular hemodynamics in remnant nephrons. AB - Endogenous glucocorticoid (GC) has been proposed to play a role in the adaptive functions of remnant nephron and participates in the progression of renal disease. The effect of GC blockade by RU-486 (20 mg/kg), an anti-GC agent, on the progression of chronic renal failure (CRF) was evaluated in Munich-Wistar rats. CRF was induced by 5/6 nephrectomy. Global renal function, glomerular hemodynamics, proteinuria and renal histopathology studies were performed after 60 days of CRF induction. RU administration in control or CRF groups did not induce significant changes in total renal function, mean arterial or intraglomerular hydraulic pressures, 24-hour proteinuria or sclerosis index. However, RU induced a significant reduction in single-nephron glomerular filtration rate in the superficial nephrons in both groups' control (decreases 20%) and CRF (decreases 57%), without changing total glomerular filtration rate, when compared with vehicle administration. These reductions were due to a decline in glomerular plasma flow rate (QA) and in glomerular ultrafiltration coefficient (Kf). These data suggest that GC played a role in the adaptive hyperfiltration associated with the compensatory mechanism but did not participate in the genesis of proteinuria or glomerulosclerosis in this experimental model. PMID- 9208282 TI - Protein kinase C beta II isoform is up-regulated in human proliferative glomerulonephritis. AB - Cell proliferation is a predominant feature in glomerulonephritis (GN). Recent work has suggested that protein kinase C (PKC) isoforms are responsible, specifically, PKC beta II in part, for cell growth. PKC beta II is expressed during cell growth in early glomerulogenesis and inflammatory mediators of glomerular disease induce PKC beta II expression. We therefore investigated the expression of PKC beta II in kidney biopsy specimens from patients with various types of proliferative (n = 41), nonproliferative GN (n = 23), and in structurally normal kidneys (n = 15). PKC beta II immunoreactivity was exclusively found in proliferative GN whereas PKC expression was not detected in normal glomerular and in nonproliferative disease states. The consistent expression of PKC beta II in proliferative GN suggests a key signaling role for this enzyme in cell proliferation in renal disease. PMID- 9208283 TI - Opposing effects of interleukin-1 and transforming growth factor-beta on the regulation of tissue-type plasminogen activator and plasminogen activator inhibitor type-1 expression by human mesangial cells. AB - Several proteases have been implicated in regulating the turnover of mesangial matrix within the glomerulus. One such group, the plasminogen activators (PAs) and their inhibitor, PA inhibitor type-1 (PAI-1), is produced by glomerular cells with altered expression during nephritis. We report the effect of IL-1 and transforming growth factor-beta (TGF-beta) and a combination of the two cytokines on the secretion of tissue-type PA (t-PA) and PAI-1 from human mesangial cells. IL-1 significantly downregulates PAI-1 production and upregulates t-PA over a range of concentrations (1-10 U/ml), while TGF-beta (0.5-10 ng/ml) has the opposite effect. The combined effect of these factors is that TGF-beta attenuates the increase in t-PA and decrease in PAI-1 expression caused by IL-1, and has a dominant effect upon their secretion, thus decreasing t-PA and increasing PAI-1. These findings contribute to our understanding of fibrinolysis and tissue remodelling within the glomerular mesangium during the course of nephritis. PMID- 9208284 TI - Competition of iron and aluminum for transferrin: the molecular basis for aluminum deposition in iron-overloaded dialysis patients? AB - In the recent literature an inverse relationship between iron status and serum aluminum levels has repeatedly been reported in dialysis patients. To check whether this observation is, at least in part, due to an interference of iron with the protein binding of aluminum, we studied the effect of the latter element on both the number of free binding sites on transferrin (Tf) and on the affinity of the protein for aluminum. For the purpose of this, a recently developed HPLC ETAAS hybrid method was used, allowing protein-binding studies at clinical relevant metal concentrations and under contamination-free conditions. After we incubated apo-Tf with iron and aluminum which were added in amounts equivalent to the calculated number of metal-binding sites on the protein (i.e., 2 mol metal/mol Tf), we found that Tf can be saturated for 100% with iron. However, for aluminum only a 23% aluminum-Tf saturation was observed. In Tf solutions with iron saturations ranging between 0 and 60% as well as in the serum of 15 subjects with iron-Tf saturations varying between 12 and 48%, a significant (p < 0.001) negative correlation between the degree of iron-Tf saturation and the percentage of aluminum (added in amounts equivalent to the number of the remaining binding sites on Tf) bound to Tf was noted (y = -0.26x + 24.5, r = -0.87 in serum). It is concluded that the iron-Tf saturation influences the Tf binding of aluminum not only by occupying binding sites otherwise available for aluminum, but also by lowering the affinity of Tf for aluminum. The effects of iron on serum aluminum levels and bone aluminum deposition are discussed. PMID- 9208285 TI - A thermodynamic equilibrium model for calcium salt urolithiasis: clinical application. AB - It is proposed that urine is better modelled as a true equilibrium rather than in a supersaturated/metastable state and that the free citrate3- ion plays a major role in maintaining dispersion of the solid particles (reduced agglomeration). Published urinary chemistries, in conjunction with the computer programme SEQUIL, have been used to formulate a novel risk index for calcium stone formation independent of the traditional clinical classification of the stone former. Applying the risk index to three consecutive 24-hour urine samples of 39 untreated Ca stone formers showed that 35 (90%) patients produced at least one abnormal urine. Traditional single urinary parameter assessment, Ca/Cr, oxalate/Cr or citrate/Cr ratios, showed that only 17, 14, and 18% of the patients, respectively, had an abnormality, while taking all three together 24 (70%) had some abnormality and thus 30% were 'idiopathic' stone formers. Treatment regimens have been devised using the computer programme to return an abnormal urine to the normal according to the proposed risk index. In most urines two or more factors had to be changed simultaneously. Clinically there has been only one recurrence in 36 months, whereas before they had 4.4/3 years. PMID- 9208286 TI - Selective antagonism of the AT1 receptor inhibits the effect of angiotensin II on DNA and protein synthesis of rat proximal tubular cells. AB - The proliferation and hypertrophy of renal tubular cells are primary features in the progression of both acute and chronic renal disease often indicating a poor prognosis. Angiotensin II (ANG II), acting alone or in combination with other growth factors, has been implicated in this process. The aims of this study were to identify the importance of both ANG II and serum-derived factors upon cellular DNA synthesis and protein synthesis in renal proximal tubular cells and to identify the roles of the ANG II receptor subtypes in these processes together with the underlying intracellular signalling mechanisms involved. Primary cultures of renal proximal tubular cells were prepared from freshly isolated rat kidney cortex. Cells were cultured in either serum-replete Dulbecco's modified Eagle's/Ham's F12 or serum-deplete defined medium containing insulin, hydrocortisone, sodium selenite, transferrin, and tri-iodothyronine. Cells were incubated with ANG II (10(-10), 10(-8), 10(-6) M) for 24-120 h either alone or in combination with losartan, PD123319, or pertussis toxin. Incubation of proximal tubular cells in the presence of serum and ANG II (10(-8) M) induced a significant early (24 h) increase in DNA synthesis, together with a significant late (96 h) increase in protein content. [3H]thymidine uptake increased by 56% (p < 0.001) and total protein content by 23% (p < 0.05). In defined media, ANG II (10(-8) M) stimulated protein synthesis only. [3H]uridine uptake, [3H]leucine uptake, and total protein content increased by 25, 57, and 17% (p < 0.05), respectively. In both serum-replete and serum-deplete media, the effects upon protein synthesis of ANG II were inhibited by pertussis toxin and losartan, but not by PD123319. ANG II is clearly a potent stimulator of renal tubular cell DNA and protein synthesis-a response mediated via the AT1 receptor coupled to a pertussis toxin sensitive Gi protein. PMID- 9208287 TI - Replacement of the lateral malleolus of the ankle joint with a reversed proximal fibular bone graft. AB - Infrequently, prior reports have described the use of the ipsilateral proximal fibula to replace an absent distal fibula caused by either trauma, infection, or resection for tumor. This is a 27-year follow-up of a 12-year-old patient who lost the distal 7.5 cm of her fibula secondary to trauma. The soft tissue defect was replaced early by an abdominal flap and the bone defect was eventually replaced with 7.5 cm of proximal fibula. The lateral ankle ligaments were reconstructed with the peroneus brevis, and the ankle joint has remained stable. Although traumatic arthrosis has progressed slowly, the patient at age 39 has a relatively painless, mobile ankle joint. PMID- 9208288 TI - Observer reliability in ankle radiographic measurements. AB - We analyzed 50 sets of ankle radiographs to determine the interobserver and intraobserver reliability when obtaining common linear and angular measurements. The radiographs were divided into two groups: one group included 25 normal ankles, and the second group included 25 fractured ankles. Each set of radiographs was evaluated independently by four different observers on two separate occasions under controlled conditions. Six radiographic parameters were measured on all 50 sets of films: syndesmosis A, syndesmosis B, syndesmosis C, the medial clear space, and the talocrural and bimalleolar angles. On the 25 sets of fracture films, four additional measurements of fracture displacement were included: displacement of the medial malleolus (mortise), displacement of the lateral malleolus (AP and lateral), and displacement of the posterior malleolus. Reliability was evaluated with an analysis of variance intraclass correlation coefficient. Among the examiners, 9 of the 10 parameters could be measured reliably. Intraobserver reliability was found to increase with the experience of the examiner. PMID- 9208289 TI - Anterior sliding graft for tibiotalar arthrodesis. AB - The results using the anterior sliding graft technique with rigid internal fixation for tibiotalar arthrodesis were reviewed. The indications for anterior sliding graft technique included posttraumatic arthritis, rheumatoid arthritis, pseudarthrosis following prior attempt at arthrodesis, and postinfectious arthrosis. The arthrodesis rate was 95%. The overall prevalence of complications was 33%. The complications related to this method were minor and easily managed. The authors concluded that the anterior sliding graft technique is performed with readily available resources, has a high rate of union, and avoids the routine use of iliac bone graft. PMID- 9208290 TI - Intrachondral microvasculature in the human fetal talus. AB - The intrachondral microvasculature of the growing talus of human was studied in 16 fetuses aged from 15 to 44 weeks of gestation, using interrupted serial sections and vascular injection of ink. The cartilage model of the talus was shown to be well vascularized throughout by cartilage canals. The cartilage canal contained blood vessels and connective tissue, with vessels originating from the perichondrial vessels. They were covered by a thick connective tissue wall that was continuous with the perichondrium. The functions of the cartilage canals were mainly to nourish the large masses of cartilage and to supply osteogenic tissue, which initiates the primary ossification center. As in the adult, the fetal talus was supplied with four to five main branches originating from the sinus tarsi and the tarsal canal; there were no anastomoses between the vessels of the adjacent cartilage canals and between the branches within the cartilage canal. This type of microvasculature is vulnerable to injury and, if impaired, may cause serious complications. PMID- 9208291 TI - Kinematic and neuromuscular changes of the gait pattern after Achilles tendon rupture. AB - After long immobilization periods in equinovalgus with operated Achilles tendon rupture, long-lasting changes of motor patterns in functional movement can be expected. In the present study, possible alterations in gait pattern have been analyzed based on kinematic and neuromuscular parameters. Ten patients 1 year after surgery and a healthy control group performed 10 gait cycles in natural walking cadence. Ankle motion, pressure distribution, and electromyographic data were recorded and analyzed in defined phases. Kinematic and neuromuscular changes are still evident 1 year after surgery with a temporal phase shift and a neuromuscular deficit of the lateral gastrocnemius muscle. The objective of rehabilitation should be the facilitation of the temporal innervation pattern of the lateral gastrocnemius muscle in the functional movement. PMID- 9208292 TI - Prevalence of radiographic foot abnormalities in patients with diabetes. AB - Clinicians are increasingly aware that mechanical aspects of foot deformities, such as Charcot changes, clawtoes, bunion deformities, or cavus or planus foot deformities, might have an impact on the occurrence, potential healing, and recurrence of foot ulcers. We report the prevalence of plain radiographic changes and attempt to rate the severity of those deformities in the feet of 456 diabetic veteran medicine clinic enrollees. All 456 radiographs were reviewed by orthopaedic surgeons to specifically identify Charcot changes, presence of arterial calcification, dislocation of the lesser toe metatarsophalangeal joints, hallux interphalangeal joint dislocation, and radiographic evidence of previous surgery. Radiographs of 428 patients were taken while weight-bearing, and these were reviewed to quantify hallux valgus angles, intermetatarsal 1-2 angles, fifth metatarsal-proximal phalangeal angles, second metatarsal lengths, lateral talocalcaneal and talar-first metatarsal angles, and claw toe deformities. The prevalence of Charcot changes was 1.4% (six subjects), and all had radiographic evidence of midfoot Charcot changes. Other deformities, such as clawtoes, hallux valgus, lesser toe joint dislocations, and alterations in arch height, are more common in veterans with diabetes. PMID- 9208293 TI - Intermittent pneumatic pedal compression and edema resolution after acute ankle fracture: a prospective, randomized study. AB - Thirty patients with an acute Weber B or C ankle fracture were enrolled after signing an informed consent. Fifteen patients were randomized to a control group where they received a posterior splint, ice, and elevation before surgery. Fifteen patients were randomized to a pneumatic pedal compression group where they received the same treatment plus a pneumatic pedal compression device which was used full-time before surgery. Baseline volumetric measurements of the injured foot were obtained, followed by measurements at 24-hour increments until surgery. On average, the patients in the pneumatic pedal compression group had an 88 mL decrease in volume in the first 24 hours versus a 33 mL increase in the control group (P < 0.03) and a 31 mL decrease in the first 48 hours of treatment versus a 32 mL increase for the control group (P < 0.05). The pneumatic pedal compression was well tolerated by the majority of patients (only one did not tolerate its use because of pain) and, we believe, serves as a useful adjunct in preoperative edema resolution after ankle fracture. PMID- 9208294 TI - Radiographic and computed tomographic evaluation of Lisfranc dislocation: a cadaver study. AB - Six cadaver feet were used for radiological and computed tomographic (CT) evaluation. The tarsometatarsal joints of each specimen were displaced dorsolaterally in successive 1-mm increments. None of the 1-mm and two thirds of the 2-mm dorsolateral Lisfranc dislocations could be visualized on routine radiographs; they could all be noted on CT scans. There was good assessment on CT scan for the extent of the minor lesions that are normally obscured by overlapping projection in routine radiographs. A Lisfranc injury that appears undisplaced on radiographs or acceptable after closed reduction may still have an unpredictable outcome because of the presence of an occult joint subluxation. CT scanning is more sensitive than radiography for detecting the minor amounts of Lisfranc displacement. If there is any doubt on the radiographs, a CT scan should be performed. The early diagnosis and treatment of Lisfranc injuries may minimize development of post-traumatic degenerative arthritis. PMID- 9208295 TI - Anatomy of the Lisfranc joint complex. AB - The authors studied the Lisfranc joint complex using gross dissection and examination of anatomical sections of frozen samples in the frontal and sagittal planes. They distinguished a medial compartment, a central compartment, a lateral compartment, the secondary joint line, and the connections with the cuneoscaphoid articulation. The ligaments were divided on the basis of topography (dorsal, interosseous, and plantar) and course (longitudinal, oblique, and transverse). The dorsal and plantar ligaments reinforce the articular capsules. The interosseous ligaments are the strongest. A common characteristic of these ligaments is that they vary considerably in course, number, and insertions. PMID- 9208296 TI - Osteochondritis dissecans of the head of the talus. AB - We present a case of osteochondritis dissecans of the head of the talus, without a history of trauma. The clinical and radiological features and a 2-year and 3 month follow-up are discussed. PMID- 9208297 TI - Subluxation of the peroneal tendons within the peroneal groove: a report of two cases. PMID- 9208298 TI - Internal fixation of the proximal chevron osteotomy. PMID- 9208299 TI - Stress fracture of the first metatarsal. PMID- 9208300 TI - Talonavicular arthrodesis for the painful adult acquired flatfoot. PMID- 9208302 TI - Distribution of endothelin 3-like immunoreactivity in gonadotrophs of the bullfrog (Rana catesbeiana) pituitary. AB - Immunohistochemical and immunocytochemical techniques were employed to investigate the distribution of endothelin 3 (ET3)-like immunoreactivity in the pituitary of the bullfrog, Rana catesbeiana. ET3-immunoreactive (ET3-IR) cells were scattered all over the pars distalis of the female pituitary; however, only a few ET3-IR cells were observed in the male pituitary. ET3-IR cells were found to correspond to cells immunostained with monoclonal antibodies against the beta subunit of bullfrog LH (fLH beta) or monoclonal antibodies against the beta subunit of bullfrog FSH (fFSH beta) at the light microscopic level. However, we could not find ET3-IR cells which were immunoreactive for other pituitary hormones. So far, all ET3-IR cells showed both fLH beta and fFSH beta immunoreactivity. About 24% of the fLH beta-IR cells and about 33% of the fFSH beta-IR cells showed ET3-like immunoreactivity. Immunoelectron microscopic analysis using colloidal gold revealed the coexistence of ET3-like substance(s) and gonadotropins within the same granules. This study demonstrated the presence of ET3-like peptide(s) in bullfrog gonadotrophs, suggesting the possible participation of ET3 in regulating pituitary function as an autocrine and/or paracrine hormone. PMID- 9208301 TI - Effect of chronic naloxone and morphine treatments on testicular, body weight, and plumage pigmentation cycle of lal munia, Estrilda amandava. AB - At Imphal (24 degrees 44' N) testes of lal munia, Estrilda amandava, began in June/July, peaked in September/ October, and thereafter declined to a minimum in December/January. Daily im treatments of 2.5-10 mg/kg/ bird/30 days of naloxone during progressive phase suppressed testicular growth, but without effects during quiescent, peak, and regression phases. Daily morphine (5 mg/kg/bird) during progressive and peak phases stimulated testicular growth, but without effects during quiescent and regression phases. Daily morphine (10 mg/kg/bird) during progressive phase stimulated the testes, an effect reversed by daily im treatments of an equivalent dose of naloxone. Seasonal changes in body weight closely correlated with testicular size. Daily im naloxone (5 and 10 mg/kg/bird/30 days) during progressive phase inhibited the increased body weight, but had no effects during quiescent, peak, and regression phases. Morphine (5 mg/kg/bird/day) during progressive and peak phases increased body weight, but had no effects during quiescent and regression phases. Morphine (10 mg/kg/bird/day) during progressive phase increased body weight, an effect which was reversed by equivalent dose of naloxone. Plumage color increased progressively between May and August/September, was maintained during October, and thereafter declined to reach dull-brown henny feathers by December. Daily im naloxone (2.5-10 mg/kg/bird/30 days) regardless of the reproductive states did not affect plumage color cycle. Morphine (5 mg/kg/ bird/day) accelerated plumage pigmentation between June and August, but had no effect during progressive or peak phases. Postnuptial decline in plumage color was inhibited by morphine (5 and 10 mg/kg/bird/day) and naloxone failed to reverse this effect. It is concluded that in the lal munia, endogenous opioid peptides are important constituents of the neuroendocrine mechanisms that influence development of the testes and body weight. PMID- 9208303 TI - Responsiveness of adenylate cyclase to pituitary gonadotropins and evidence of a hormone-induced desensitization in the lizard ovary. AB - Gonadotropins (FSH and LH) affect several mammalian gonadal functions. In particular, FSH stimulates oogonial proliferation and oocyte growth, while LH regulates ovulation and progesterone secretion. In lacertilian reptiles, gonadal function is also regulated by pituitary gonadotropins, but which hormone controls ovarian activities and the mechanisms of action are unknown. The present study aimed to clarify mechanisms of action of pituitary gonadotropins on the ovary of Podarcis sicula (Lacertilia). The data demonstrate that mammalian gonadotropins FSH and LH produce a threefold stimulation of adenylate cyclase activity in follicular membranes, while hCG and TSH are less effective, causing a twofold increase in adenylate cyclase activity. Neurotransmitters such as dopamine, serotonin, and catecholamines have no effect on enzyme activity. The action of mammalian FSH and LH on the ovary mimics the effect of homologous hormones: in lizard ovaries incubated in vitro in the presence of isolated homologous pituitary glands, the intracellular cAMP level increased by 50% with respect to control ovaries. Mammalian gonadotropins appear homologous to lizard gonadotropin(s): Southern blot analyses show that the lizard genome contains nucleotide sequences homologous to those encoding for mammalian beta FSH and beta LH. Both homologous and heterologous desensitization of adenylate cyclase activity occurs in the lizard ovary. In fact, responsiveness of adenylate cyclase to gonadotropin stimulation is abolished in animals 2 hr after in vivo treatment with FSH. Sensitivity to gonadotropin stimulation is restored 2 weeks after the beginning of the in vivo treatment. Desensitization was also observed in ovaries incubated in vitro with mammalian FSH or with isolated pituitary glands. PMID- 9208304 TI - The effects of N-methyl-D,L-aspartate and gonadotropin-releasing hormone on in vitro growth hormone release in steroid-primed immature rainbow trout, Oncorhynchus mykiss. AB - In the present study we investigated the effects of 17 beta-estradiol (E2) and 5 alpha-dihydrotestosterone (5 alpha DHT) on the ability of the glutamate agonist, N-methyl-D,L-aspartate (NMA), to stimulate growth hormone (GH) release from perifused pituitary glands of sexually immature rainbow trout (Oncorhynchus mykiss). Two weeks after steroid hormone implantation, pituitary glands were removed from the fish and challenged with NMA (10(-8) M) in a perifusion unit. NMA rapidly and significantly elevated GH release from perifused pituitary fragments taken from all treatment groups, and there was a main effect of in vivo steroid hormone treatment on the in vitro GH response to NMA. To examine the relationship between NMA and gonadotropin-releasing hormone on GH release, pituitaries from E2- and 5 alpha DHT-primed and control fish were exposed to a single pulse of salmon gonadotropin-releasing hormone (sGnRH) which also elicited a significant elevation in GH release from perifused pituitary fragments, although the response in the E2- and 5 alpha DHT-primed fish was significantly smaller (P < 0.05) than that for the NMA challenge. Administration of a specific GnRH antagonist, D-pGlu1,D-Phe2,D-Trp3,6-LHRH, did not affect the GH response to NMA, whereas administration of the NMDA receptor antagonist DL-2-amino-5 phosphonovaleric acid blocked the GH response to NMA. These data suggest that NMA acts to stimulate GH release directly at the level of the somatotroph, likely through the NMDA receptor and not through increased release of GnRH. PMID- 9208306 TI - In vitro synthesis of 17,20 beta,21-trihydroxy-4-pregnen-3-one by ovaries of turbot (Scophthalmus maximus L.) during oocyte maturation. AB - Ovaries from female turbot (Scophthalmus maximus L.), a serial spawner, were incubated in vitro with 17-hydroxy[1,2,6,7-3H]progesterone or [7 3H(N)]pregnenolone (P5-3H) during the spawning season. Several metabolites comigrated on TLC and HPLC with known reference steroids and were identified after chemical reaction and crystallization. Incubation with P5-3H generated many 4-ene steroids, accounting for 55% of total radioactivity, indicating strong 3 beta-HSD activity. The major steroids produced by ovaries were testosterone, androstenedione, and 17,21-dihydroxy-4-pregnene-3,20-dione. In addition, 17,20 alpha-dihydroxy-4-pregnen-3-one was also identified, while small quantities of 17,20 beta,21-trihydroxy-4-pregnen-3-one (20 beta-S) (maximum 1.7% of the total radioactivity) were also synthesized. The identity of 20 beta-S was confirmed in incubates with nonlabeled 17-hydroxyprogesterone by mass spectrometry. Production of 17,20 beta-dihydroxy-4-pregnen-3-one was not apparent. PMID- 9208305 TI - Effects of day length and temperature on gonadal development, body mass, and fat depots in white-crowned sparrows, Zonotrichia leucophrys pugetensis. AB - We tested the effects of ambient temperature (5 degrees, 20 degrees, and 30 degrees) on photoperiodically induced reproductive functions in male and female white-crowned sparrows, Zonotrichia leucophrys pugetensis. Transfer from short days (9L 15D) to long days (16L 8D) resulted in rapid testicular growth and partial ovarian development in all three temperature treatments. There were no differences in sizes of testes and cloacal protuberance following 30 or 70 days of exposure to long days at the different temperatures. However, brood patch and follicular development were enhanced in females at 30 degrees compared with the 5 degrees and 20 degrees groups. Many of these females exposed to 30 degrees had large yolky follicles by Day 70. This enhancement was evident only when females were housed in the same room with males, however. Despite the effects of high temperature on ovarian development, there were no differences among groups in plasma levels of follicle-stimulating hormone or luteinizing hormone, suggesting that differential ovarian development may have been mediated by gonadal sensitivity to gonadotropins rather than by differential secretion of these hormones. We examined circulating levels of corticosterone (B) and both tri iodothyronine (T3) and thyroxine (T4) as possible regulators of this differential ovarian sensitivity to gonadotropins. Plasma B levels showed transitory increases in males at 5 degrees and 20 degrees, but were suppressed in males at 30 degrees. Titers of B were not influenced by temperature treatments in females. Circulating T4 increased following photostimulation in both sexes, but this increase was reduced at 5 degrees. T3 concentrations in plasma were highly variable and not influenced by either photo-period or temperature in males, but were significantly lower in females exposed to 30 degrees by Day 70. Thus, B and T4 levels do not appear to help explain differential ovarian development, but circulating T3 levels cannot yet be excluded as a regulator of ovarian sensitivity to gonadotropins. Long days resulted in no change, or a gradual decrease, in body mass and fat deposit in males and females, and temperature regimes had no further effects on fattening or body mass. Thus, reproductive development under long days appears to be resistant to naturally relevant temperature extremes in male Z.l. pugetensis, whereas follicular development (i.e., yolk deposition in follicles leading to ovulation and onset of nesting) can be enhanced by high temperature. Reasons for the dimorphism in this response are unknown, but may be explained by the role of females in determining onset of final ovarian maturation and nesting in relation to favorable environmental conditions. In a second experiment, in which the sexes were isolated from one another, we determined the effects of the same treatments on.Z.I. pugetensis. Again there was no effect of temperature on photoperiodically induced testicular growth, and the enhancement of follicular development in females at 30 degrees was greatly reduced in the absence of males. We also continued this experiment up to 116 days of treatment to investigate effects on onset of photorefractoriness (spontaneous gonadal regression) and onset of prebasic moult. In both sexes it was clear that low temperature (5 degrees) retarded gonadal regression and high temperature (30 degrees) advanced it. Similarly, the prebasic moult score was greater at 30 degrees and less at 5 degrees in both sexes. There were no effects of temperature on plasma levels of LH at Day 116 of treatment, but plasma levels of T4 were higher in the 5 degrees group of both males and females sampled at Day 116. Clearly, the effects of temperature can have different effects on gonadal recrudescence, onset of breeding (yolk deposition), and termination of breeding. Whether these influences of temperature on reproductive function at different stages in the breeding cycle have different mechanisms remains to be determ PMID- 9208307 TI - Individual diurnal plasma profiles of thyroid hormones in rainbow trout (Oncorhynchus mykiss) in relation to cortisol, growth hormone, and growth rate. AB - In order to characterize the individual diurnal plasma profiles of triiodothyronine (T3) and thyroxine (T4) in rainbow trout (Oncorhynchus mykiss), blood samples from 41 fish were taken every hour during a 24-hr period, through a catheter inserted into the dorsal aorta. The possible influences of day-night alternation, sex, and diet (feed intake, time of meals) on thyroid hormone (TH) profiles were analyzed. The existence of relations between diurnal plasma profiles of T3, T4, T3/T4 ratio, and those of the growth hormone (GH), cortisol (previously described in Gomez et al., J. Exp. Zool. 274, 171-180, 1996), and the growth rate was monitored. Average daily T3 and T4 concentrations were, respectively, 2.6 +/- 0.2 and 5.5 +/- 0.3 ng/ml (n = 41). Our study showed little or no variation in plasma T3 concentrations during one 24-hr period, while those of T4 fluctuated markedly. T4 peaks occurred from a baseline of 4.0 +/- 0.2 ng/ml at a frequency of 2.5 +/- 0.2 peaks/24 hr, with an amplitude of 3.0 +/- 0.4 ng/ml, and a duration of 4.3 +/- 0.4 hr. There was a significant difference between the average circulating T3 level during the day and that at night (2.4 +/ 0.2 vs 2.7 +/- 0.2 ng/ml). No influence of sex or food factors was observed on daily TH concentrations. TH peaks occurred irregularly and asynchronously without apparent influence of day-night alternation, sex, and diet. The growth rate was significantly correlated with the daily T3 concentration (r = 0.77), but not with T4. No significant relationships were found between daily concentrations of T3, T4, GH, and cortisol. The absence of a relationship between TH and GH concentrations suggests that, in salmonids, GH may have no observable short-term action on the conversion of T4 to T3. PMID- 9208308 TI - Action of different ecdysteroids on the regulation of mRNAs for the ecdysone receptor, MHR3, dopa decarboxylase, and a larval cuticle protein in the larval epidermis of the tobacco hornworm, Manduca sexta. AB - To determine which ecdysteroids may be biologically active in the larval epidermis of the tobacco hornworm, Manduca sexta, we studied the action of several known ecdysteroids and metabolites on the expression of the genes encoding the ecdysone receptor (EcR), Manduca hormone receptor 3 (MHR3), dopa decarboxylase (DDC), and a larval cuticle protein (LCP-14). Both Day 2 fourth- and Day 2 fifth-instar larval epidermis contained significant 3 beta-reductase activity which metabolized 3-dehydroecdysone (3DE) and 3-dehydro-20 hydroxyecdysone (3D20E) to ecdysone (E) and 20-hydroxyecdysone (20E), respectively, but had only very low amounts of ecdysone oxidase activity (E to 3DE) and no detectable ecdysone 20-monooxygenase activity (E to 20E). When the expression of the various genes was studied in the epidermis in vitro, 20E and 3D20E had similar effects, whereas E, 3DE, 26-hydroxyecdysone and 20,26 dihydroxyecdysone were ineffective. Exposure of Day 2 fifth-instar epidermis to 500 ng/ml of either 20E or 3D20E for 24 hr caused a rapid, biphasic increase in EcR-B1 mRNA. By contrast, EcR-A mRNA showed a less rapid initial increase followed by a slow steady rise and was less responsive to 3D20E. Ecdysone in a 1:1 mixture with 20E effectively halved the concentration of 20E needed to induce EcR-B1 mRNA but showed no synergism in the induction of EcR-A mRNA. The induction of MHR3 mRNA and of DDC mRNA in Day 2 fourth-instar epidermis as well as the suppression of DDC and LCP-14 gene expression by 3D20E was indistinguishable from that of 20E. Therefore, for Manduca larval epidermis, only 20E and 3D20E are biologically active ecdysteroids. Since the 3D20E can be converted to 20E by the epidermis, its effects are likely mediated by 20E. PMID- 9208310 TI - Ontogeny of insulin-like growth factors (IGF-I and IGF-II) and IGF-binding proteins in the chicken following hatching. AB - The present study examined plasma concentrations of insulin-like growth factor (IGF)-I, IGF-II and IGF-binding proteins (IGFBPs) during posthatch growth and development in chickens. Three distinct proteins which bound 125I-IGF-I were observed irrespective of age or sex, these having apparent molecular weights of 22, 28, and 36 kDa. The major IGFBP present during much of the growth and development period was the 28-kDa form followed by the 36-kDa form. Plasma concentrations of IGF-II and of the 22-kDa IGFBP showed little ontogenic variation with the exception of elevated levels of the 22 kDa IGFBP in 1-day-old chicks. The circulating concentrations of IGF-I and of the 28-kDa IGFBP increased progressively between the time of hatching to reach a maximum at 6 weeks of age and subsequently declined to lower levels in adults. Somewhat similarly, the 36 kDa IGFBP increased during early pre- and posthatching growth to a maximum at 6 weeks of age. There were marked sex differences in circulating concentrations of IGF-I in young (4 week) and adult chickens and in the 36-kDa IGFBP in the adult, both being lower in females. Estrogen treatment of adult male chickens decreased the circulating concentrations of IGF-I together with the level of both the 28- and 36-kDa IGFBPs. Testosterone treatment had no effect on the circulating concentrations of either IGF-I or IGFBPs in adult female chickens. We conclude that the relative levels of IGF-I, IGF-II, and the IGFBPs change with age. In addition, circulating concentrations of estrogen may play a role in the regulation of IGF-I and IGFBPs. PMID- 9208309 TI - Quantification of salmon alpha- and thyrotropin (TSH) beta-subunit messenger RNA by an RNase protection assay: regulation by thyroid hormones. AB - In order to study salmon thyroid-stimulating hormone (TSH), we designed a highly specific, sensitive, and rapid RNase protection assay (RPA) for quantification of steady-state levels of salmon TSH beta-subunit mRNA expression. The cDNA encoding the beta-subunit of TSH was isolated from coho salmon pituitary total RNA by reverse-transcriptase PCR, partially sequenced, and used as template for synthesizing a radioactively labeled, sequence-specific, antisense probe, and sense standard for the RPA. This assay, along with a similar RPA previously designed for coho salmon total alpha-subunit mRNA, was used to examine the effects of feeding T3 (0, 10, 100 micrograms/g) and methimazole (a thyroid inhibitor) (2.5 mg/g) on TSH subunit gene expression after 2 and 4 weeks. The low dose of T3 (10 micrograms/g) caused no change in TSH beta mRNA after 2 and 4 weeks and a transient increase in alpha mRNA after 2 weeks, followed by no significant effect after 4 weeks. The high dose of T3 (100 micrograms/g) caused a decrease in TSH beta mRNA after 4 weeks and no change in total alpha mRNA after 2 and 4 weeks. In contrast, methimazole treatment caused significant increases in both TSH beta mRNA (250%) and alpha mRNA (50%) levels after 4 weeks. These findings confirm that, as in mammals, TSH alpha- and beta-subunit expression in teleosts may be differentially regulated by negative feedback from the thyroid hormones. PMID- 9208311 TI - Metabolism of estrogens and androgens by embryonic tissues of Arctic charr, Salvelinus alpinus. AB - This study examines the ability of Arctic charr (Salvelinus alpinus) embryos to metabolize tritiated androstenedione (A4), testosterone (T), 17 beta-estradiol (E2), and estrone (E1) in vitro; the metabolic products were separated by HPLC. A4 was poorly metabolized, with 48 to 64% of the substrate remaining even after 24 hr of incubation. The major metabolites of A4 metabolism are E1 and some other unidentified metabolites. T was mostly converted to A4, along with some reduced steroids, but E2 was a minor metabolite. Further, while E2 was almost exclusively transformed into E1, when E1 was used as the precursor, there was little metabolism; the products of E1 metabolism were small amounts of E1 sulfate, glucuronide, E2, and an unknown metabolite which cochromatographed with reference steroid androstenetrione (also called 11-ketoandrostenedione). It is concluded that in Arctic charr embryos there is preferential expression of a form of 17 beta-hydroxysteroid dehydrogenase resembling the type 2 isozyme of mammals that converts T and E2 to A4 and E1, respectively. PMID- 9208312 TI - Enhancement by prolactin of the GnRH-induced release of LH from dispersed anterior pituitary cells of the bullfrog (Rana catesbeiana). AB - The response of enzymatically dispersed anterior pituitary cells of the bullfrog (Rana catesbeiana) to gonadotropin-releasing hormone (GnRH) was studied by monitoring the release of luteinizing hormone (LH) into the culture medium. The cells responded to GnRH by releasing LH according to the incubation time and to the GnRH concentration. The responsiveness to GnRH became less conspicuous as the cell density was reduced. Addition of prolactin (PRL) to the medium enhanced the responsiveness to the secretagogue, and addition of antiserum against PRL lowered the responsiveness to a certain extent. Immunohistochemical studies of sectioned pituitaries revealed that PRL cells most frequently located in contact with LH cells. The possibility that PRL acts directly on gonadotrophs to enhance their responsiveness to GnRH was suggested. PMID- 9208313 TI - Sexual bipotentiality of developing ovaries in the terrestrial isopod Armadillidium vulgare (Malacostraca, Crustacea). AB - The androgenic glands (AG) of crustaceans are responsible for differentiation of male sexual characters. The process of gonadal differentiation in females was studied morphologically in Armadillidium vulgare given a masculinizing AG implant. Gonadal masculinization was induced by implantation of an AG into females at various stages of postembryonic development. Functional sex reversal always occurred when an AG was implanted into females that were in stages 5 and 6 of development. Partial formation of testes was induced after implantation of an AG into stage 7 and 8 females. When an AG was implanted into a stage 9 female, development of a functional testis was not observed, but the ovaries were partially masculinized. These results show that after the onset of sex differentiation female gonads retain sexual bipotentiality through several stages of postembryonic development. Implantation of one AG into a female is enough to induce gonadal masculinization and sex reversal in this species. The AG implant up to stage 6 (3.4 mm in body length) is an experimental procedure certain to transform a genetic female into a functional male. The process of gonadal development in female A. vulgare is discussed. PMID- 9208314 TI - [Transurethral incision for ureterocele in children]. AB - Transurethral incision (TUI) was performed as the initial treatment in 10 children with ureteroceles. Three patients had ureteroceles associated with a single ureter. TUI relieved hydronephrosis and preserved renal function in all 3 cases. Urinary tract infection developed in no patients. However, all the patients required an antireflux operation because of postoperative vesicoureteral reflux (VUR). Seven children had a total of 8 ureteroceles associated with a duplex system. TUI resulted in preservation of the upper pole function in 6 of the 8 ureteroceles. Urinary tract infections and VUR developed in 3 and 7 patients, respectively. Common sheath reimplantation was performed in 2 ureteroceles. TUI relieves obstruction before the onset of devastating infections although it carries the risk of postoperative VUR. We recommend TUI as the initial treatment for ureteroceles associated with both single and duplex systems. PMID- 9208315 TI - [Clinical outcome of bladder preserving therapy for superficial high-grade bladder cancer]. AB - Clinical outcome of bladder preserving therapy for superficial high-grade bladder cancers (pTa-pT1b, G3) was retrospectively studied. Twenty-six patients initially received bladder preserving therapy ('preserved' group), while 19 underwent radical cystectomy as the initial treatment ('cystectomized' group). Tumor recurrence was accompanied by a progression in the tumor grade in 9 patients with low-grade tumors ('grade-up' group). The 5-year survival rate was 87% and 100% in the preserved and cystectomized groups, respectively, with no significant difference. However, the grade-up group had a significantly worse rate of 57%. Failure in bladder preserving therapy was unrelated to the size or number of tumors. Our findings suggest that bladder preserving therapy is a reasonable treatment option for superficial high-grade tumors although a careful followup is mandatory. On the other hand, radical cystectomy should be considered for recurrent, grade-up G3 tumors. PMID- 9208316 TI - [Inhibitory effect of oral administration of Takusya on calcium oxalate crystallization in human whole urine]. AB - We previously reported that Takusya had the inhibitory effect on in vitro calcium oxalate crystallization and in vivo stone formation in an animal model and it could be a prophylactic agent against calcium oxalate stone formation. We studied the effect of Takusya on calcium oxalate crystallization in human urine. Takusya (500 mg/day and 1,000 mg/day) was administered to 16 healthy men for 3 days and then 24-hour urine samples were collected to measure the urinary excretion of calcium, phosphate, magnesium, uric acid, creatinine, citric acid and oxalic acid. The urine samples before the administration of Takusya was used as a control. The size and distribution of crystals, which were formed in the urine samples by adding calcium chloride and sodium oxalate, were measured using the Coulter counter technique. Urinary magnesium excretion was significantly reduced by 1,000 mg/day of Takusya compared with the control (p < 0.05). The growth of crystals was significantly inhibited by 500 mg/day of Takusya in the large crystal formers whose urine created crystals more than 3.5 microns before the administration of Takusya (p < 0.05). These findings suggested that Takusya inhibited the growth of crystals formed in human urine. PMID- 9208317 TI - [A case of erythropoietin-producing renal cell carcinoma proved by immunohistochemistry]. AB - A case of erythropoietin-producing renal cell carcinoma is reported. A 59-year old woman was referred to our hospital for further examination of erythrocytosis accompanied with an elevated serum erythropoietin level. Ultrasonography and computerized tomography scan showed a left renal tumor. She underwent a left radical nephrectomy. Postoperatively, serum erythropoietin level and erythrocytosis were normalized. Histopathology was renal cell carcinoma, clear cell subtype, and immunohistochemistry revealed erythropoietin in cancer cells. Furthermore, the cancerous tissue contained a high concentration of erythropoietin. PMID- 9208318 TI - [Renal cell carcinoma associated with renal artery aneurysm]. AB - We report the 5th case of renal cell carcinoma associated with renal artery aneurysm in the Japanese literature. A 57-year-old woman was admitted to our hospital for further examination of a right renal mass. Computerized tomography and magnetic resonance imaging revealed a 6 x 5 cm tumor at the upper pole of the right kidney and an aneurysm of the right renal artery. An angiography showed duplicated renal arteries and a saccular aneurysm 2.1 x 2.5 cm occurring at the first bifurcation of one renal artery. A transperitoneal radical nephrectomy was performed. PMID- 9208319 TI - [Metastatic renal tumor originating from esophageal cancer: report of 2 cases]. AB - We report 2 cases of esophageal cancer metastatic to the kidney. The first case was in a 57-year-old man who complained of severe right flank pain. He had underwent an operation for esophageal cancer 2 months previously. A computerized tomography (CT) scan revealed a wedge-shaped, low-density mass in the right kidney. Right nephrectomy revealed squamous cell carcinoma. He has remained free of recurrence 3 months postoperatively. The second case was in a 57-year-old man with esophageal cancer treated by radiation therapy. Severe right flank pain and gross hematuria appeared after 1 year. A CT scan showed a wedge-shaped, low density tumor in the right kidney accompanied with a tumor thrombus in the inferior vena cava. Right nephrectomy as well as resection of the thrombus were performed. Pathological diagnosis was squamous cell carcinoma. He died of cancer 2 months postoperatively. PMID- 9208320 TI - [Wilms' tumor with marked calcification in an 8-year-old boy: a case report]. AB - An 8-year-old boy presented with asymptomatic gross hematuria. Clinical investigation revealed an 8-cm left renal tumor accompanied with marked calcification. Radical nephrectomy with lymph node dissection was performed. Pathological diagnosis was Wilms' tumor without lymph node metastasis. He has been free of recurrence 20 months postoperatively. Our case features prominent calcification on radiological examination, which is uncommon in Wilms' tumor. We reviewed the literature on the relationship between renal tumors and calcification. PMID- 9208321 TI - [Adenocarcinoma of the bladder producing carcinoembryonic antigen and carbohydrate antigen 19-9: report of a case]. AB - We report a case of primary adenocarcinoma of the bladder producing carcinoembryonic antigen (CEA) and carbohydrate antigen 19-9 (CA19-9). An 85-year old woman presented with gross hematuria. Computerized tomography and ultrasonography demonstrated a mass on the urinary bladder. After clinical examination, she was diagnosed with locally invasive bladder cancer. Serum CEA and CA19-9 levels were increased to 8.9 ng/ml (normal < 2.5) and 870 U/ml (normal < 37), respectively. Simple cystectomy and bilateral cutaneous ureterostomy were performed. The histopathological diagnosis was well-differentiated adenocarcinoma of the bladder with invasion through the entire muscle layer. Postoperatively, the serum CEA and CA19-9 levels decreased to 5.4 ng/ml and 53 U/ml, respectively, and both CEA and CA19-9 were detected by immunohistochemical examination of the tumor. PMID- 9208322 TI - [Bladder cancer in patients with spinal cord injury: report of two cases]. AB - Two cases of bladder cancer in patients with spinal cord injury are reported. A 57-year-old paraplegic woman who had had an indwelling urethral catheter for 20 years visited our hospital with macroscopic hematuria. A bladder tumor was found endoscopically. A transurethral biopsy revealed a squamous cell carcinoma. She refused operation and received radiation therapy. The second case was in a 69 year-old paraplegic man on clean intermittent catheterization for 2 years presenting with a high grade fever. The diagnosis was a transitional cell carcinoma of the urinary bladder. Bilateral cutaneous ureterostomy was performed because of tumor invasion. He received radiation therapy but died of cancer 18 months after surgery. PMID- 9208323 TI - [A case of vesical endometriosis associated with Wunderlich syndrome]. AB - A case of vesical endometriosis associated with unilateral renal agenesis and uterus didelphys (Wunderlich syndrome) is reported. A 20-year-old unmarried woman visited the gynecologic department with dysmenorrhea. She was referred to us because a bladder mass was detected on abdominal ultrasonography. She had no episode of gross hematuria. Cystoscopy revealed a finger tip-sized cystic mass dark red to dark blue in color. Computerized tomography and magnetic resonance imaging demonstrated right renal agenesis and uterus didelphys. The mass was located at the ureterovesical pouch and protruded into the bladder. Partial cystectomy, right hysterectomy and right salpingo-oophorectomy were performed. Histopathological diagnosis was endometriosis. The patient was treated with 400 mg danazol for 6 months has had no recurrence and regular menstruation. PMID- 9208324 TI - [A case of urachal abscess resected electively after successful drainage from umbilicus]. AB - A 22-year-old woman presented to our hospital complaining of high fever and a painful fist-sized mass in the lower abdomen. From the results of imaging studies, including abdominal urtrasonography and computerized tomography, the patient was diagnosed as having a urachal abscess. Despite intensive antibiotic therapy, her high fever persisted. Drainage via the umbilicus was then performed with a pigtail catheter being inserted into the abscess through a fistula. Her temperature normalized within a few days and the size of the abscess gradually decreased. Total excision of the urachus and partial cystectomy were carried out 19 days after the drainage. The drainage procedure allowed us to perform surgery with relative case and to reduce the extent of the tissue to be excised. PMID- 9208325 TI - [Malignant lymphoma of the penis: report of two cases]. AB - We report two cases of malignant lymphoma of the penis. A 64-year-old man presented with painful indurations of the penis. A computerized tomography (CT) scan showed a swollen retroperitoneal lymph node 5 cm in diameter. Penectomy confirmed the diagnosis of B cell malignant lymphoma, diffuse large type. Despite systemic chemotherapy with a CHOP regimen, he died of disease 7 months postoperatively. The second case was in a 63-year-old man presenting with multiple nodules in the penis and scrotum. Biopsy of a scrotal nodule revealed B cell malignant lymphoma, diffuse medium size type. A CT scan demonstrated widespread lesions (stage III E). He has been in complete remission for 10 months following, multidisciplinary treatments. We reviewed 21 cases of malignant lymphoma of the penis which have been reported in the literature. PMID- 9208326 TI - Internalization of ecto-ATPase activity in human neutrophils upon stimulation with phorbol ester or formyl peptide. AB - We have demonstrated the alteration of the localization of ecto-ATPase activity in human neutrophils after stimulation with phorbol myristate acetate or N formylmethionyl-leucyl-phenylalanine using a cerium-based cytochemical method. In unstimulated cells, the enzyme activity was observed on the plasma membrane. Both the diazonium salt of sulfanilic acid and diethylpyrocarbonate inhibited the production of the reaction precipitates. Within 2-3 min of stimulation, cells developed cytoplasmic projections (ruffles). The ecto-ATPase activity on the plasma membrane of these ruffles was, however, weaker than that at the non-ruffle forming side. The ruffle-forming side (RFS) was also the site where elongated tubular structures positive for the enzyme reaction tended to concentrate and associated with the plasma membrane. The enzyme activity was also detected in intracellular compartments, which appeared predominantly in the RFS concomitantly with the disappearance of the enzyme activity from the plasma membrane. Using a series of thick sections and computer-assisted three-dimensional reconstruction, the enzyme reaction-positive internalized membranes were visualized as a complicated mass formed by enzyme reaction-positive vesicles which gathered together and were, at least in part, interconnected. The present results indicate that the detected enzyme reaction is a product of the ecto-ATPase activity, and that RFS possibly serves the membrane flow with respect to endocytosis. PMID- 9208327 TI - Expression of p53 protein and p53 gene transcripts in rabbit carotid arteries after balloon denudation. AB - Smooth muscle cell (SMC) proliferation may be positively or negatively regulated by various factors after arterial wall injury. We investigated the hypothesis that the p53 protein may play a role in regulating SMC proliferation. We examined p53 protein expression and p53 gene transcript distribution in rabbit carotid arteries over a period of 6 weeks after balloon denudation in relation to SMC proliferation. The number of p53-positive SMCs in the neointima reached a maximum 2 weeks after balloon denudation when proliferating SMCs, decreased and were confined to the luminal surface of the intima. The p53-positive SMCs in the neointima almost paralleled the distribution of proliferating cell nuclear antigen (PCNA)-positive SMCs but not that of Ki-67-positive cells. Double immunofluorescence showed the simultaneous nuclear localization of both p53 protein and PCNA in intimal SMCs. Strong signals for p53 gene transcripts, identified by in situ hybridization, were observed in SMCs showing positive immunostaining for p53 protein. Our results indicate that p53 protein and p53 gene transcript levels increase, are closely linked to the proliferation of SMCs in the thickened intima, and play a key role in the regulation of cell proliferation during the repair process after arterial wall injury. PMID- 9208328 TI - Lysozyme expression in the rat parotid gland: light and electron microscopic immunogold studies. AB - Lysozyme (muramidase) is capable of direct bacteriolytic action by hydrolyzing glycosidic bonds in bacterial cell walls. Although it is broadly distributed in vertebrate tissues and secretions, the cellular and subcellular localizations of the enzyme are still not well known. The present study examines the distribution of lysozyme expression in the various cell types of LR gold-embedded rat parotid gland, applying a postembedding immunogold-silver staining technique for light microscopy. Simultaneously, a postembedding immunogold method for electron microscopy was used to determine the cellular compartments engaged in the biosynthesis and exocytosis of lysozyme. Silver-amplified immunogold staining for lysozyme demonstrated identical localization in both paraffin and semithin LR gold sections: in the supranuclear parts of acinar and intercalated duct cells. Staining intensity varied even between adjacent cells. In the electron microscope, immunogold labeling was detected over the cell compartments associated with protein synthesis and exocytosis in acinar and intercalated duct cells. Lysozyme antigenic sites were visible over endoplasmic reticulum and throughout the Golgi apparatus, being intense over the trans-Golgi network, but even stronger in the condensing vacuoles and most prominent over secretory granules in both cell types. The findings provide the first immunocytochemical evidence of the synthesis and secretion of lysozyme in parotid acinar and intercalated duct cells. PMID- 9208329 TI - Mercurochrom can be used for the histochemical demonstration and microphotometric quantitation of both protein thiols and protein (mixed) disulfides. AB - Mercurochrom [2,7-dibromo-4-(hydroxymercuri)-fluorescein disodium salt] used for staining of protein thiols in addition binds to other groups of proteins. Experimental evidence is provided that mercurochrom bound to non-thiol groups forms a 1:1 adduct with protein (mixed) disulfides. The disulfide contents of three different types of cells determined biochemically correlated with the corresponding mean integrated optical densities determined microphotometrically after mercurochrom staining of groups other than thiols. Intracellular disulfide exchange has been studied, leading to a transformation of protein mixed disulfides to protein disulfides and an equimolar loss of protein thiols. Protein mixed disulfides were generated from protein thiols using both methyl methanethiosulfonate (MMTS) and 2,2'-dihydroxy-6,6'-dinaphthyldisulfide (DDD). Loss of thiols as well as the equimolar increase of protein mixed disulfides were followed using both mercurochrom staining for thiols and for disulfides. Generation of protein mixed disulfides due to the DDD reaction was also followed by azocoupling with Fast blue B. On the basis of the observed stoichiometry between the loss of protein thiols and the quantity, increase or conversion of protein disulfides determined microphotometrically using both mercurochrom staining and DDD Fast blue B staining, we conclude that: (1) 1 mol of mercurochrom is bound per mol of protein (mixed) disulfide; and (2) the molar absorptivity of mercurochrom bound to disulfides is epsilon 520 = 34940. This study demonstrates that mercurochrom can be used for the quantitative determination of the oxidative status of protein thiols in cells. PMID- 9208330 TI - Identification of phagocytic glial cells after lesion-induced anterograde degeneration using double-fluorescence labeling: combination of axonal tracing and lectin or immunostaining. AB - Retrograde and anterograde degeneration have been reported to be sufficient stimuli to activate glial cells, which, in turn, are involved in phagocytosis of degenerating material. Here we describe a double-fluorescence technique which allows for direct and simultaneous visualization of both labeled incorporated axonal debris and incorporating glial cells in the course of anterograde degeneration. Stereotaxic application of small crystals of biotinylated and tetramethylrhodamine (TRITC)-conjugated dextran amine Mini Ruby into the medial entorhinal cortex resulted in a stable rhodamine fluorescence confined to fibers and terminals in the middle molecular layer of the dentate gyrus, the stratum lacunosum-moleculare, and the crossed temporo-hippocampal pathway. Subsequent stereotaxic lesion of the entorhinal cortex induced transformation of rhodamine fluorescent fibers and terminals into small granules. Incorporation of these granules by microglial cells [labeled by fluorescein isothiocyanate (FITC) coupled Bandeiraea simplicifolia isolectin B4] or astrocytes (labeled by FITC coupled glial fibrillary acidic protein antibodies) resulted in phagocytosis dependent labeling of these non-neuronal cells, which could be identified by double-fluorescence microscopy. Electron microscopical analysis revealed that, following lesion, the tracer remained confined to entorhinal axons which were found to be incorporated by glial cells. Our data show that TRITC- and biotin conjugated dextran amines are versatile tracers leading to Phaseolus vulgaris leucoagglutinin-like axonal staining. Lesion-induced phagocytosis of anterogradely degenerating axons by immunocytochemically identified glial cells can be directly observed by this technique on the light and electron microscopical levels. PMID- 9208331 TI - Heterogeneity of liver cells expressing procollagen types I and IV in vivo. AB - The extracellular matrix of fibrotic liver is predominantly produced by mesenchymal cells, the in vivo phenotype of which is poorly defined. We report on the application of combined immunohistology and in situ hybridization with [35S] labeled alpha 1(I) and alpha 1(IV) procollagen RNA probes. In CCl4-treated rats, all alpha 1(I) procollagen-producing cells were vimentin positive but cytokeratin negative; over 90% expressed desmin, a marker of rat liver stellate cells. alpha 1(I) Procollagen-expressing, desmin-negative cells were confined to portal tract and perivascular stroma. Similarly, alpha 1(I) procollagen gene transcripts were, in all instances, colocalized with vimentin in human liver. In fibrotic specimens, over 70% of these cells expressed alpha-actin. Antibodies against epithelial, endothelial, and Kupffer cells, granulocytes, and lymphocytes did not react with alpha 1(I) procollagen RNA-expressing cells. Localization, morphology, and immunophenotype of alpha 1(I) procollagen-expressing cells indicated stellate cells and portal/vascular fibroblasts, but not epithelial cells, to be sources of hepatic interstitial collagen. However, most alpha 1(IV)-expressing human liver cells were identified as endothelial cells, the remaining cells were (myo )fibroblastic and bile duct epithelial cells, but not hepatocytes. This indicates synthesis of procollagen type IV from both sides of the basement membrane and suggests an active participation of endothelial cells in the process of sinusoidal capillarization. PMID- 9208332 TI - In situ assessment of mRNA accessibility in heterogeneous tissue samples using elongation factor-1 alpha (EF-1 alpha). AB - Elongation factor-1 alpha (EF-1 alpha) is an evolutionarily highly conserved universal cofactor of protein synthesis in all living cells. In this study, its use as a positive control in situ hybridization assays for specific detection of mRNA sequences was evaluated. Northern blot analysis of various non-neoplastic and neoplastic cultured cells of different stages of confluence, cell shape, and cell cycle status revealed that EF-1 alpha had a lower and more homogeneous expression than did beta-actin. In situ hybridization assays using digoxigenin labeled riboprobes for the detection of EF-1 alpha mRNA in routinely formalin fixed, paraffin-embedded tissue sections showed that EF-1 alpha is a suitable positive control in all types of cells. However, variation of protease pretreatments demonstrated distinct and sometimes mutually exclusive digestion conditions for different cell types within the same tissue sample. Our results indicate that detection of EF-1 alpha mRNA is an appropriate internal standard for in situ hybridization assays and that it is useful to control artifacts such as false negatives caused by inappropriate protease pretreatments. The observed variability of optimal protease pretreatments for different cell types within the same tissue section strengthens the importance of a positive control in in situ hybridization assays. PMID- 9208333 TI - Redistribution of carbonic anhydrase VI expression from ducts to acini during development of ovine parotid and submandibular glands. AB - Carbonic anhydrase VI (CA VI) is a secreted enzyme produced predominantly by serous acinar cells of submandibular and parotid glands. We have investigated the developmental pattern of CA VI production by these glands in the sheep, from fetal life to adulthood, using immunohistochemistry. Also, a specific radioimmunoassay for CA VI was used to measure changes in enzyme expression in the parotid gland postnatally. CA VI is detectable by immunohistochemistry in parotid excretory ducts from 106 days gestation (term is 145 days), in striated ducts from 138 days and in acinar cells from 1 day postnatal. The duct cell content of CA VI declined as the acinar cell population increased, a feature also of CA VI immunoreactivity in the submandibular gland. Production of CA VI by submandibular duct cells was detectable initially at 125 days gestation, and acinar production was not seen before 29 days post-natal. Apart from the differing ontogeny of CA VI production in ducts and acini of parotid and submandibular glands, there was a parallel pattern of CA VI expression during the development of these major salivary glands. With the development of the acinar tissues in the postnatal lamb, there was a dramatic increase (about 600-fold) in the level of expression of CA VI in the parotid gland between days 7 and 59 as measured by radioimmunoassay. PMID- 9208335 TI - Once-daily dosing of aminoglycosides. A consensus document. PMID- 9208334 TI - Human neutrophil lipocalin (HNL) is a specific granule constituent of the neutrophil granulocyte. Studies in bronchial and lung parenchymal tissue and peripheral blood cells. AB - The neutrophilic granulocyte is a cytotoxic and potentially tissue-injuring cell participating in the destructive processes and symptoms seen in a variety of inflammatory diseases. Sensitive immunoassays have been introduced to measure the levels of specific secretory proteins of various inflammatory cells in blood and other body fluids. The aim has been to develop highly specific markers for each cell type. The results obtained by immunoassay have indicated that human neutrophil lipocalin (HNL) is a protein unique to the neutrophil. The present study investigated the specificity of HNL as a neutrophil marker in peripheral blood and lung tissue by using flow cytometry and immunocytochemistry. Flow cytometry and immunocytochemistry on peripheral blood showed that monoclonal antibodies to HNL only react with neutrophils and not with other types of leukocytes. Immunocytochemistry on plastic-embedded sections and on frozen sections of lung tissue showed that a cocktail of six monoclonal antibodies to HNL specifically reacts with neutrophils and not with epithelial cells or macrophages. By immunoelectron microscopical studies performed on healthy human neutrophils after low temperature embedding in Lowicryl K4M following aldehyde fixation and partial dehydration, it could be shown that HNL colocalized with lactoferrin (a known marker for secondary or specific granules) and that myeloperoxidase was localized in the primary or azurophil granules. The results confirm that HNL is a unique component of the secondary granules of the neutrophil granulocyte. PMID- 9208336 TI - Metabolic effects of spirapril and atenolol: results from a randomized, long-term study. AB - Thirty-seven patients with mild to moderate hypertension were randomized in a double-blind, parallell-group study to receive either 6 mg of spirapril or 50 mg of atenolol. After 6 weeks of treatment, the daily dose of spirapril was increased to 12 mg and the daily dose of atenolol to 100 mg, if the target diastolic pressure < or = 90 mmHg was not achieved. The total treatment period was 12 months for both drugs. There were no changes in the lipid levels in the spirapril treatment group whereas the concentration of apoprotein A1 decreased significantly (p < 0.05) in the group treated with atenolol when compared with baseline. There was a tendency in the levels of serum cholesterol and triglycerides to increase and in the level of HDL cholesterol to decrease in the atenolol treatment group compared with baseline. Fasting blood glucose levels did not change significantly in the treatment group when compared with baseline. The 2-hour blood glucose level in the glucose tolerance test increased significantly after the 12-month therapy with both spirapril (p < 0.05) and atenolol (p < 0.05) when compared with baseline levels. There was also a tendency in the 2-hour insulin levels to increase in both the spirapril and the atenolol treatment group after the 12-month treatment period. In conclusion, spirapril had no effect on lipid metabolism whereas atenolol had some untoward effects. In the group treated with atenolol fasting blood glucose levels increased, but both spirapril and atenolol increased the 2-hour blood glucose levels in the glucose tolerance test. PMID- 9208337 TI - The effects of antihypertensive combination therapy on lipid and glucose metabolism: hydrochlorothiazide plus sotalol vs. hydrochlorothiazide plus captopril. AB - Metabolic side-effects of antihypertensive drugs may increase the risk of coronary heart disease despite an adequate blood pressure reduction. Since combinations of different antihypertensive drugs are often necessary and frequently used, we performed a randomized study comparing the effects of a fixed combination of hydrochlorothiazide and sotalol (group A), or hydrochlorothiazide and captopril (group B) on blood pressure and on lipid and glucose metabolism in 40 men with essential hypertension over 1 year. Significant blood pressure reductions (p < 0.001) were achieved in both treatment groups: from 160/105 to 128/88 mmHg in group A (mean doses: hydrochlorothiazide 33 and sotalol 197 mg) and from 162/106 to 135/89 mmHg in group B (hydrochlorothiazide 33 and captopril 64 mg) after 12 months, respectively. No significant changes in body weight were observed in either treatment group. Triglycerides increased (p < 0.05) in both treatment groups (from 183 to 262 mg/dl in A, and from 160 to 196 mg/dl in B) and HDL cholesterol decreased (p < 0.001 and < 0.05) in both groups (from 45.1 to 35.7 mg/dl in A, and from 49.3 to 46.3 mg/dl in B), whereas LDL cholesterol increased significantly (p < 0.05) only in group A from 153 to 164 mg/dl. No significant changes were observed in total cholesterol nor in lipoprotein(a) concentrations in either treatment group. Fasting plasma glucose and hemoglobin A1 increased significantly (p < 0.05) only in group A after 1 year of treatment (from 91.6 to 98.0 mg/dl, and from 6.3 to 6.9%, respectively). Serum levels of creatinine and potassium decreased, and uric acid increased significantly under either combination. Our data show that the diuretic/beta-blocker combination has adverse effects on lipid and glucose metabolism after long-term therapy. The effects of the diuretic/ACE inhibitor combination on lipid metabolism are less pronounced and there are no adverse effects on glucose metabolism. However, the ACE inhibitor component could not completely counteract the metabolic effects of the diuretic. Both combinations have no effects on Lp(a). We conclude that the combination of hydrochlorothiazide with an ACE inhibitor has a better metabolic profile for the treatment of essential hypertension than the combination with a beta-blocker. PMID- 9208339 TI - Pharmacokinetics of incadronate, a new bisphosphonate, in healthy volunteers and patients with malignancy-associated hypercalcemia. AB - We investigated the pharmacokinetics of incadronate, a new bisphosphonate, after a 2-hour intravenous infusion to healthy volunteers and patients with malignancy associated hypercalcemia. Following administration at 0.025-1.6 mg to healthy volunteers, peak plasma concentration of incadronate increased in a dose proportional manner. Plasma concentration thereafter declined biexponentially with a half-life of 0.26-0.40 h (t1/2 alpha) and 1.58-1.98 h (t1/2 beta). AUC increased dose-proportionally, whereas distribution volume (Vdss), total body clearance (CLt), renal clearance (CLr) and nonrenal clearance (CLnr), corresponding to bone uptake clearance, changed little among doses, indicating the linear pharmacokinetics of the drug after intravenous administration. Within 24 h, 55.1-69.5% of the dose was excreted into urine as the unchanged drug, most in the first 6 h. CLr and CLnr accounted for about 60% and 40% of the CLt, respectively, suggesting that the pharmacokinetics of incadronate are affected by changes in these clearances. In patients, plasma concentration at 2 h increased dose-proportionally in the range of 2.5-10.0 mg. Urinary excretion of YM175 up to 24 h after dosing was as low as 10.5% of the dose, being 1/6 of those in volunteers. A positive correlation (r > 0.89) was observed between creatinine clearance and urinary incadronate excretion in all volunteers and patients, indicating that the reduction of urinary excretion in patients is due to the decrease in renal function accompanying hypercalcemia. Based on comparison of the dose-normalized plasma concentration, the plasma level at 2 h in patients was comparable with that in volunteers, whereas the level at 8 h was 3 times higher in patients, suggesting that elimination from plasma in patients is delayed due to decreased renal function. Nevertheless, the plasma concentration profile in patients was lower than that predicted from the decrease in CLr. This finding suggests that the increase in plasma concentration with decreasing renal excretion in hypercalcemic patients was compensated for by enhanced bone uptake of the drug. PMID- 9208338 TI - Endothelial cell compatibility of fluoroquinolone solutions for intravenous use. AB - One local side-effect closely related to the use of parenteral fluoroquinolones is phlebitis. The occurrence of this phenomenon is largely thought due to the damage of endothelial cells with subsequent inflammation. In order to evaluate the effect of ciprofloxacin, fleroxacin, and ofloxacin on the viability of human umbilical venous endothelial cells (HUVEC), intracellular ATP levels were measured by a luciferin-luciferase assay. Prostacyclin (PGI2) and thromboxane A2 (TXA2) were determined by means of direct radioimmunoassay. Commercially available preparations of ciprofloxacin (2 mg/ml) and fleroxacin (4 mg/ml) reduced the intracellular ATP content by 75.9 +/- 1.9% and 82.1 +/- 0.6%, respectively, within 20 minutes, indicating severe damage of endothelial cells. Incubation with ofloxacin (2 mg/ml) did not have any detrimental effect. All fluoroquinolones were tolerated well by endothelial cells at low concentrations up to 20 micrograms/ml. Concentrations between 100-200 micrograms/ml gradually led to functional alterations such as increased PGI2 release. The tolerance of intravenously applied antibiotics has been tested in animal models. Use of human venous endothelial cells for testing antibiotic solutions for intravenous application provides a valuable alternate model for tolerability. PMID- 9208340 TI - Use of flurythromycin ethylsuccinate in infections of lower airways due to sensitive germs: multicenter comparative study with clarithromycin. AB - We have conducted a trial according to double-blind, double-dummy conditions, comparing flurythromycin ethylsuccinate with clarithromycin treatments. Sensitive pathogenic bacteria were isolated according to the Kirby Bauer method from sputum cultures of 152 patients affected by acute infections of lower airways (bronchitis, bronchopneumonia, etc.). The activities of the 2 drugs were investigated on the basis of a main bacteriological evaluation and of several secondary criteria; the 2 treatments were effective and tolerated to the same extent. It can be concluded that in the treatment of infections of lower respiratory airways the use of flurythromycin offers some advantages. PMID- 9208341 TI - Influence of cholestyramine on the pharmacokinetics of cerivastatin. AB - The possible influence of the bile acid sequestering agent cholestyramine, a basic comedication in hypercholesterolemic patients, on the pharmacokinetics of the new HMG-CoA reductase inhibitor cerivastatin was investigated. When both drugs were administered concomitantly in the morning under fasting conditions, a decrease in relative bioavailability by 21% could be observed, possibly due to irreversible adsorption of the statin to the resin. In addition, the delay in absorption led to a 41% decrease in cerivastatin mean maximum plasma concentration which also occurred at later time. A second study addressed in detail the question of time interval required between both treatments to minimize the influence of cholestyramine pretreatment on cerivastatin bioavailability: dosing of cerivastatin at dinner (6 p.m.) or bed time (10 p.m.) with cholestyramine pretreatment 1 hour before meal (5 p.m.) in both treatments. The decrease in mean AUC was now approximately 8-16% depending on the time of pretreatment (1-hour-interval: 16%, 5-hour-interval: 8%), and Cmax decreased by approximately 32%, irrespective of the time of pretreatment. Tmax was increased in both treatments, whereas t1/2 was not changed. The presented data support the conclusion that when administered concomitantly, the bioavailability of cerivastatin is moderately reduced by adsorption to cholestyramine. Following, however, the dosing instructions of both cholestyramine (1 hour before meal) and cerivastatin (once-daily in the evening at dinner or at bed time), i.e. administering both drugs several hours (at least 1 hour) apart, the observed effects on rate and extent of absorption of cerivastatin are unlikely to be of clinical relevance. PMID- 9208342 TI - Absolute and relative bioavailability of the HMG-CoA reductase inhibitor cerivastatin. AB - To determine the absolute bioavailability of the HMG-CoA reductase inhibitor cerivastatin, 12 healthy young male volunteers received single doses of either 100 micrograms as a 1-minute bolus infusion or 200 micrograms orally as tablets in a controlled, randomized crossover study. In addition, 8 of the 12 subjects participated in a third treatment period in which 200 micrograms cerivastatin were administered as an oral solution as reference for determining the relative bioavailability of the tablet drug formulation. Plasma samples were analyzed for cerivastatin by a specific HPLC assay with fluorescence detection after post column irradiation of the eluate, with a limit of quantification of 0.1 microgram/l. Following all treatments, cerivastatin was well tolerated and no clinically relevant adverse events or changes in laboratory parameters were observed. Vital signs and ECG remained unchanged. Plasma concentration/time profiles of cerivastatin following intravenous bolus could be described by a 2 compartment model with a distribution half-life of 3-5 min and an elimination half-life of 1.5-2.4 h. For the 2 oral administrations a 1-compartmental pharmacokinetic model with a first-order absorption process was best to describe the plasma concentration/time data. Based on the AUCnorm values of the 7 subjects, valid for complete pharmacokinetic evaluation, the absolute bioavailability of tablet and oral solution was 60.0 and 59.6% (90% confidence intervals 53-68%), respectively. The relative bioavailability of tablet compared with solution was 100.7% (90% confidence interval 89-114%), with tablet and oral solution showing nearly identical in vivo absorption characteristics and almost superimposable plasma concentration/time curves. The tablet formulation, therefore, can be regarded as an optimal oral formulation with respect to galenic aspects. PMID- 9208344 TI - To the psychological effects of youth unemployment: international perspectives. PMID- 9208343 TI - Influence of the antacid Maalox and the H2-antagonist cimetidine on the pharmacokinetics of cerivastatin. AB - The possible influence of Maalox 70, an antacid based on magnesium-aluminum hydroxide, and the H2-antagonist cimetidine, both commonly prescribed in hypercholesterolemic patients, on the pharmacokinetics of the new HMG-CoA reductase inhibitor cerivastatin was investigated in 2 separate studies in 8 healthy young male subjects each. Cerivastatin plasma concentration/time profiles were assessed by a specific HPLC assay; in addition, total immunoreactive drug (cerivastatin plus metabolites) was determined by RIA. Single oral doses of 200 micrograms cerivastatin were administered under fasting conditions without or with 10 ml Maalox 70 suspension. The mean AUC and Cmax ratios (combined dosing/monodosing) including 90% confidence intervals were 0.92 (0.73-1.15) and 0.89 (0.72-1.10) for the HPLC data, and 0.99 (0.85-1.14) and 1.03 (0.82-1.30) for the RIA data, respectively. Thus, no interaction of the simultaneous administration of Maalox 70 on the pharmacokinetics of cerivastatin was observed. In a similar controlled, randomized nonblind 2-way crossover design the influence of the H2- antagonist and well-known cytochrome P450 enzyme inhibitor cimetidine was investigated. Eight healthy young male volunteers received single oral doses of 200 micrograms cerivastatin alone or on the fourth day of a 4-day cimetidine 400 mg b.i.d. pretreatment. The mean AUC and Cmax ratios (combined dosing/monodosing) including 90% confidence intervals were 0.98 (0.90-1.08) and 0.91 (0.78-1.07) for the RIA data, and 0.89 (0.82-0.96) and 0.93 (0.80-1.09) for the HPLC data, respectively, clearly indicating that cimetidine and cerivastatin did not interact pharmacokinetically. These results do not only reflect the apparent insensitivity of cerivastatin absorption to possible changes in gastric pH, but demonstrate that the metabolic pathways of cerivastatin, involved in its first-pass metabolism and elimination, are rather insensitive to cytochrome P450 enzyme inhibition induced by cimetidine. PMID- 9208345 TI - Effect of underemployment on school-leavers' self-esteem. AB - This study explores whether self-esteem is adversely affected by economic underemployment as defined by unemployment, involuntary part-time employment, intermittent unemployment, and poverty income in a group of recent school leavers. Results indicate that self-esteem was significantly lower in each of the economically underemployed groups relative to the adequately employed after controlling for early self-esteem, socio-economic status, gender, ethnicity, aptitude, age, and education. There were no differences in self-esteem among the economically underemployed groups after adjusting for the control variables. Economic underemployment proved to be a distinct concept relative to self reported job satisfaction. Underemployment was negatively related to self-esteem after controlling for perceived job satisfaction and the other control variables. Our findings suggest a need for societal attention to the levels of underemployment on par with the attention given to monitoring traditional unemployment levels. PMID- 9208346 TI - Long-term unemployment amongst adolescents: a longitudinal study. AB - The unique developmental period of adolescence suggests a distinct experience of unemployment compared to older and adult populations. A longitudinal study of long-term unemployed adolescents found healthy levels of negative self-esteem associated with strong adult social identification, high perceived time filled, low employment commitment and strong perceived personal identity. The evidence suggested good psychological health (indicated by negative self-esteem and GHQ 12) contributed to gaining employment. Employment was not associated with an improvement in levels of self-esteem (both positive and negative), although the psychological benefits of employment were indicated by a lowering in GHQ-12 scores (despite lower levels reported during unemployment by those later employed). PMID- 9208347 TI - Youth unemployment and mental health: some Dutch findings. AB - Two hypotheses were investigated: (1) the causation hypothesis that assumes that unemployment leads to poor mental health and (2) the selection hypothesis that assumes that poor mental health reduces the likelihood of finding a job. A prospective longitudinal design was used in order to study two Dutch samples: 635 college graduates and 767 school-leavers. The causation hypothesis was confirmed for school-leavers but not for college graduates. In addition, as expected, employment and further education increased levels of mental health among school leavers. The selection hypothesis, that unfortunately could only be studied in the graduate sample, was not confirmed as far as mental health was concerned. However, it appeared that future employment among graduates was predicted by a positive attitude and an active way of dealing with unemployment. Results are interpreted with reference to the favourable Dutch structural and cultural context that existed at the time the research was conducted. In addition, the role of proactivity is discussed. PMID- 9208348 TI - Nervous and depressive symptoms in a longitudinal study of youth unemployment- selection or exposure? AB - One thousand and sixty young people were followed for 5 years from the last term of compulsory school. Unemployment correlated positively with changes in nervous complaints and depressive symptoms, even after controlling for initial psychological health and background factors. There were no pronounced gender differences. Qualitative methods were used to study mediating factors between unemployment and mental health, including lack of self-confidence, self-blame, stress, isolation, lack of control and resignation. PMID- 9208349 TI - Youth unemployment and psychological distress in the Republic of Ireland. AB - Unemployment is the most significant influence on the levels of psychological distress of young adults. Unlike the situation for the adult population, social class and income are not contributory factors. Social class of origin, however, does have a contributory effect. Feelings of lack of control and attribution of responsibility for employment solely to structural or political factors increase the impact of unemployment. Evidence in relation to employment commitment does not support "culture of poverty" type explanations. Unemployed youth appear to be "people with a problem" rather than "problem people". PMID- 9208350 TI - Learned helplessness or expectancy-value? A psychological model for describing the experiences of different categories of unemployed people. AB - Various studies have explored the relationships between unemployment and expectation of success, commitment to work, motivation, causal attributions, self esteem and depression. A model is proposed that assumes the relationships between these variables are moderated by (a) whether or not the unemployed individual is seeking a first job and (b) age. It is proposed that for the unemployed who are seeking their first job (seekers) the relationships among these variables will be consistent with expectancy-value theory, but for those who have had a previous job (losers), the relationships will be more consistent with learned helplessness theory. It is further assumed that within this latter group the young losers will experience "universal helplessness" whereas the adult losers will experience "personal helplessness". PMID- 9208351 TI - International perspectives on youth unemployment and mental health: some central issues. PMID- 9208352 TI - The pharmacologic approach to the treatment of obesity. AB - Obesity is a major risk factor for morbidity and mortality, and a series of pharmacologic approaches are available for helping to manage the problem. Obesity is caused by an imbalance between caloric intake and energy expenditure, which is influenced by both environmental and genetic factors. Pharmacologic treatments include anorexigenic agents, which fall into two broad categories: those that act via brain catecholamine pathways and those that act via serotonin pathways. The most recent oral agents approved are dexfenfluramine, which is currently being marketed, and sibutramine. Both agents inhibit the control reuptake of serotonin but in addition may have effects on thermogenesis. Under investigation are agents that increase energy expenditure: the beta 3-adrenergic receptor agonists and drugs that prevent the intestinal absorption of free fatty acids and cholesterol. In development are innovative approaches to influence leptin and its receptors, various obesity genes, and biologic substances thought to influence satiety (neuropeptide Y, enterostatin, cholecystokinin, bombesin, and amylin). Obesity has now become a major target for drug development not only for affecting obesity per se but also for managing and preventing comorbid conditions such as diabetes and cardiovascular disease. PMID- 9208353 TI - Development and evaluation of a clinical psychopharmacology educational curriculum. AB - Clinical psychopharmacology training at the University of Toronto (Toronto, Ontario, Canada) was in need of improvement. The authors developed and evaluated a Clinical Psychopharmacology Educational Curriculum to complement the education provided to trainees and staff. The program consisted of journal clubs, case rounds, and didactic research tutorials and lectures. The program was attended by staff, fellows, psychiatry residents, graduate pharmacology students, medical students, and pharmacists. No course credit or continuing medical education credits were offered for participating in the program. The program ran for two academic years (September 1994-June 1996). Anonymous evaluation forms were collected after the sessions. These evaluated content, presentation style, and educational value, each on a scale from 1 (unsatisfactory) to 5 (excellent). Ratings for each component of the curriculum were close to excellent. The case rounds were rated slightly better for presentation style and educational value. Positive ratings suggest that similar efforts, particularly using case-based methods, should be developed in other teaching hospitals contingent on availability of qualified faculty and funding. PMID- 9208354 TI - Time of peak drug concentration after a single dose and a dose at steady state. AB - The time of peak concentration after administration of oral drug is an often quoted and used pharmacokinetic parameter. It is not well appreciated, however, that the peak times after a single dose and a dose at steady state during a multiple administration regimen can differ significantly. This article derives the mathematical relationships that determine how a peak time at steady state differs from that after a single or first dose. These relationships are then evaluated using three different approaches: 1) graphic simulations of time courses of drug concentration for three hypothetical drugs; 2) comparisons of predicted and observed peak times using examples from the literature; and 3) comparisons of predicted and simulated peak times based on different sampling schedules for three hypothetical drugs. The key finding is that peak times after a dose at steady state can occur considerably earlier after administration than after a single dose. However, the manner by which peak times are usually determined, that is, the sampling time corresponding to the highest measured drug concentration, imposes significant limitations on the usefulness of this parameter. PMID- 9208355 TI - Stability and performance of a population pharmacokinetic model. AB - This study aimed to determine the stability (in terms of covariate selection) of a population pharmacokinetic model and evaluate its performance in the absence of a test data set. Data from 88 full-term infants, 11 of whom were human immunodeficiency virus (HIV)-seropositive, taking an antiinfective agent were analyzed using exploratory data analysis methods and the nonlinear mixed-effects modeling (NONMEM) program to obtain the final population pharmacokinetic model. The stability of the population pharmacokinetic model was tested using the nonparametric bootstrap approach in four steps: 1) with the base pharmacokinetic model, 100 bootstrap replicates of the original data were generated by sampling with replacement; 2) ascertainment that each bootstrap data replicate was described by the basic structural model using the NONMEM objective function; 3) generalized additive modeling (GAM) applied to empiric Bayesian estimates for covariate selection at alpha = 0.05 and a frequency (f) cutoff value of 0.50; and 4) NONMEM population model building using covariates selected in the third step with alpha = 0.005. Performance of the population pharmacokinetic model was evaluated using 200 additional bootstrap replicates of the data by fitting the model obtained in step 4 to them. Parameters obtained were compared with those obtained in the model stability step, and improved prediction error, a measure of predictive accuracy as an index of internal validation, was computed. The reciprocal of serum creatinine (RSC; f = 0.73) and HIV (f = 0.70) were selected by GAM as predictors of clearance (Cl). The population pharmacokinetic model obtained without the determination of model stability included RSC as a predictor of Cl, but the final model from the model stability step included both HIV and RSC as predictors of Cl. Final population pharmacokinetic parameters were obtained with this model fitted to the original data; however, the 95% confidence interval on the HIV status regression coefficient included zero, indicating no significance. The mean parameter estimates obtained with the additional 200 bootstrap replicates of data were within 15% of those obtained with the final model at the regression stability step. Bootstrap resampling procedure is useful for evaluating the stability and performance of a population model by repeatedly fitting it to the bootstrap samples when there is no test data set. PMID- 9208356 TI - Short-term safety and efficacy of low-dose simvastatin in elderly patients with hypertensive hypercholesterolemia and fasting hyperinsulinemia. AB - To evaluate the short-term safety and efficacy of low-dose (10 mg) simvastatin in hypercholesterolemic, hypertensive elderly Chinese patients receiving antihypertensive treatment, a randomized, double-blind, placebo-controlled, 3 month trial was conducted. The patients had a total plasma cholesterol level of at least 250 mg/dL and had been, for at least 3 months, consuming a standard lipid-lowering diet (American Heart Association Step 1 Diet). Elderly hypertensive patients (n = 76) were randomized to receive treatment with either placebo (n = 38) or simvastatin (n = 38). The dosage consisted of 10 mg simvastatin daily during the 3-month trial. During that period, in the simvastatin group, plasma levels of total cholesterol and low-density lipoprotein cholesterol decreased significantly (27% and 33%, respectively) compared with those levels in plasma in the placebo group. The plasma levels of high-density lipoprotein cholesterol increased (7%), whereas triglyceride levels slightly decreased (8%). There were no serious side effects, and simvastatin was generally well tolerated. Fasting hyperinsulinemia also improved (-21%) after 3 months of simvastatin therapy. Results of this study confirmed that a low dose (10 mg) of simvastatin daily is a safe and effective method of reducing plasma levels of total and low-density lipoprotein cholesterol in hypercholesterolemic, hypertensive elderly patients receiving concurrent antihypertensive drug therapy, and that it has the additional potential benefit of reducing plasma levels of insulin. PMID- 9208357 TI - Pharmacokinetics of pirmenol enantiomers and pharmacodynamics of pirmenol racemate in patients with premature ventricular contractions. AB - The pharmacokinetics and pharmacodynamics of pirmenol were investigated in 12 patients with premature ventricular contractions (PVCs) after oral administration of racemic pirmenol, 100 mg and 200 mg every 12 hours. Holter monitoring was performed and serial blood samples were collected after the seventh doses. Plasma concentrations of pirmenol enantiomer were determined using a stereospecific liquid chromatographic assay. Clearance of total (-)-pirmenol was 20% higher than that of total (+)-pirmenol, and the difference in unbound clearance was 45% between enantiomers. Total pirmenol showed a smaller difference because of stereoselective protein binding, with 25% (100-mg dose) or 27% (200-mg dose) higher fraction unbound for (+)-pirmenol than for (-)-pirmenol. Distribution volume was similar for both enantiomers. Dose-dependent clearance was observed for unbound pirmenol enantiomers, as both enantiomers showed 20% lower unbound clearance at the higher dose. Antiarrhythmic effect (% reduction in PVCs from baseline) was correlated with plasma concentrations of pirmenol using a sigmoid maximum drug effect model, and patients showed a large variability in their antiarrhythmic response to plasma concentrations of pirmenol. The median value for minimum effective plasma concentration of racemic pirmenol was 1.5 micrograms/mL. PMID- 9208358 TI - Comparison of enalapril to captopril by 24-hour ambulatory blood pressure monitoring. AB - The efficacy of a once-daily dose of enalapril was compared with a thrice-daily dose of captopril in an open-label, randomized parallel group study of 27 hypertensive patients. The patients were monitored using conventional measurements of blood pressure and with 24-hour ambulatory blood pressure monitoring at baseline and after 12 weeks of therapy. The end points were 24 hour, daytime, and nighttime mean blood pressure values and the percentage of elevated systolic and diastolic measurements, reflecting the "hypertensive load." Enalapril reduced mean 24-hour systolic blood pressure by 18 mmHg and diastolic blood pressure by 11 mmHg. The comparative values for captopril were 9 mmHg and 2 mmHg, respectively. The mean daytime systolic blood pressure was reduced by 20 mmHg with enalapril versus 7 mmHg with captopril; the diastolic values were lowered by 11 mmHg with enalapril versus 4 mmHg with captopril. The mean nighttime systolic blood pressure was lowered by 16 mmHg with enalapril versus 12 mmHg with captopril; the diastolic values were reduced by 10 mmHg with enalapril and 5 mmHg with captopril. No major side effects were recorded in either group. A single daily 20-mg dose of enalapril, therefore, proved to be equipotent or superior to 75 mg of captopril administered in three divided doses. PMID- 9208359 TI - Steady-state pharmacokinetic properties of pramipexole in healthy volunteers. AB - Pramipexole is a dopamine receptor agonist that has proved effective in the treatment of Parkinson's disease. The pharmacokinetic properties of pramipexole at steady-state concentrations were studied in 16 healthy men and women at four dose levels throughout the range recommended for Parkinson's patients. Plasma and urine samples collected within the four dose intervals were assayed for concentrations of pramipexole, using high-performance liquid chromatography. The total oral clearance for all participants was 419 mL/min. The mean volume of distribution and elimination half-life for all participants was 486 +/- 93.2 L and 12.9 +/- 3.27 hours. Concentrations of pramipexole were proportional to dose, although the drug's pharmacokinetic properties differed between men and women. The area under the concentration-time curve for each dose level was 35% to 43% greater in women, mainly because of a 24% to 27% lower oral clearance. The mean creatinine clearance in men and women was 112 +/- 12.8 mL/ min/1.73 m2 and 80.9 +/- 15.6 mL/min/1.73 m2, respectively. The renal clearance of pramipexole accounts for approximately 80% of oral clearance, and there was a significant correlation between renal and creatinine clearances. The influence of gender could not be distinguished from the influence of age and the resulting reduced creatinine clearance, but the measurement of pharmacokinetic properties produced linear results in both men and women. PMID- 9208360 TI - Postabsorption concentration peaks with brand-name and generic verapamil: a double-blind, crossover study in elderly hypertensive patients. AB - The pharmacokinetic actions, bioequivalence, and cardiovascular effects of two verapamil products were studied in a randomized, double-blind, crossover study in eight elderly hypertensive patients (median age, 69.5 years; range, 60-79 years) given brand-name or generic immediate-release verapamil in 120-mg twice-daily doses for 14 days. Blood pressures, heart rates, P-R intervals; and serum concentrations of R-/S-verapamil and norverapamil were measured multiple times in patients during the last day of each therapy. Median blood pressure decreased more with generic verapamil than with the brand-name drug, with the largest difference occurring at 0.5 hours (137/74 mmHg versus 144.5/80.5 mmHg; P = 0.05 and 0.091, respectively). Pharmacokinetic parameters were not different for the two products (P < 0.01). However, the generic product, compared with the brand name drug, had mean area under the concentration-time curve (time 0 to 12 hours) ratios (90% CI) of 1.09 (0.78-1.52), 1.16 (0.87-1.55) and 1.11 (0.81-1.52) for R , S-, and total verapamil. Seventy concentration peaks (31 with the brand-name drug, 39 with the generic drug) appeared between 8 and 24 hours. Median percentages of increase of these peaks, compared with those of previous concentrations, were 48.3% and 36.3% for brand-name and generic drugs, respectively. Fifty of the 70 peaks (71%) were associated with a stereospecific concentration peak of norverapamil and, temporally, with meals. Our findings suggest that whereas the two verapamil products may not be bioequivalent by Food and Drug Administration criteria, the observed differences in effects were not clinically significant in this elderly population. Multiple concentration peaks after absorption were observed in all patients with both verapamil products and were perhaps related to enterohepatic recirculation. PMID- 9208361 TI - Effect of a cola beverage on the bioavailability of itraconazole in the presence of H2 blockers. AB - Thirty-three healthy individuals participated in an open-label, randomized, three way crossover study designed to compared the bioavailability of a single 200-mg oral dose of itraconazole when administered alone or after treatment with ranitidine, both with and without coadministration of a cola beverage. Each treatment phase was separated by a 2-week washout period. Participants pretreated with ranitidine were required to have a gastric pH of at least 6.0 before receiving itraconazole. An analysis of the area under the curve (AUC) and peak plasma concentration (Cmax) data indicated that the bioavailability of itraconazole was significantly reduced when the gastric pH was increased by pretreatment with ranitidine but showed that this effect was counteracted by the coadministration of an acidic solution (e.g., a cola beverage) that transiently reduced the gastric pH. These findings suggest that the coadministration of an acidic beverage with itraconazole may be an effective approach in improving the bioavailability of itraconazole in patients who are hypochlorhydric or who are taking gastric acid suppressants. PMID- 9208362 TI - Epidemiology: an essential science for speech-language pathology and audiology. AB - This article introduces epidemiology as a health science that is essential as a complement to the basic laboratory and clinical sciences in speech-language pathology and audiology. A contemporary definition of epidemiology is presented. Principles of epidemiology, including causal criteria, and use and misuse of concepts such as incidence, prevalence, and risk are elaborated from the perspective of practitioners and researchers in human communication sciences and disorders. The ubiquity of epidemiologic data in the news media illustrates the complexities of data interpretation. PMID- 9208363 TI - Epidemiology as an essential tool for establishing prevention programs and evaluating their impact and outcome. AB - Emerging trends in health care place speech-language pathologists and audiologists on the threshold of role expansion. To include prevention efforts within this new role, clinicians need to ask about the determinants of communication disorders that are preventable within the general population or within sub-populations. In this article, epidemiology is presented as a tool for the primary prevention of a broad range of factors associated with increased rates of communication disorders. "Old" epidemiologic theory, based on populations, blends with the "new" epidemiologic approach emphasizing the contribution of individual behaviors to adverse health outcomes, to provide the working clinician with a framework from which to determine the direction prevention efforts must take. The PRECEDE-PROCEED model of program planning and evaluation (Green and Kreuter, 1991) is utilized in public health and holds potential for use by clinicians in preventing communication disorders. Application of communication disorders to the PRECEDE-PROCEED model is offered as an example of prevention planning. PMID- 9208364 TI - Epidemiologic issues in audiology: impact on professional training and service delivery. AB - Universal infant hearing screening and the transition of audiology from a master's to a doctoral level profession are discussed from an epidemiologic point of view. Epidemiologic strategies which may inform decision-making in these two important and controversial arease are presented. PMID- 9208365 TI - Epidemiologic principles in studies of infectious disease outcomes: pediatric HIV as a model. AB - Epidemiologic principles have been employed in the investigation of AIDS since the early 1980s. Although such principles have demonstrated the difficulties in reporting the everchanging rates of incidence and prevalence, in addition to distributions of children with HIV, they have also established specific pieces of a multifaceted puzzle. Professionals interested in examining only a piece of the puzzle, such as a particular communication disorder, often are unable to see how it fits into the complete puzzle. This article presents several epidemiologic findings of pediatric HIV, including population distributions, a summary of modes of transmission, occurrence of opportunistic infections, and manifestations of the disease in child populations. It also discusses HIV-related speech, language, swallowing, and voice disorders, examining the complexities of quantification of risk for each piece within the pediatric HIV puzzle. The purpose is to broaden the perspective of professionals concerned with how these disorders fit within the overall puzzle of immunocompromised populations of children. PMID- 9208366 TI - Epidemiology of specific language impairment: prenatal and perinatal risk factors. AB - The prenatal and perinatal risk factors likely to be associated with specific language impairment (SLI) were examined in this study. A review of existing research showed that there have been few studies on this topic. Among children with SLI, greater rates of near relatives with language learning problems has been found. Data obtained from a case-control study of 177 children with SLI and 925 children without sensory, developmental disorders, or language impairment were studied using a parental questionnaire concerning exposures during the prenatal and perinatal period for the index children. Differences between the children with SLI and controls were found for parental characteristics regarding education, positive history of language and learning problems, tobacco smoking, and breast feeding. No support was found for elevated rates of maternal exposure to disease or occupational chemical substances. PMID- 9208367 TI - Characterising non-covalent interactions with the Cambridge Structural Database. AB - This review describes how the CSD can be used to study non-covalent interactions. Several different types of information may be obtained. First, the relative frequencies of various interactions can be studied; for example, we have shown that the terminal oxygen atoms of phosphate groups accept hydrogen bonds far more often than the linkage oxygens. Secondly, information can be obtained about the geometries of nonbonded contacts; for example, hydrogen bonds to P-O groups rarely form along the extension of the P-O bond, whereas short contacts between oxygen and carbon-bound iodine show a strong preference for linear C-I ... O angles. Thirdly, the CSD can be searched for novel interactions which may be exploited in inhibitor design; for example, the I ... O contacts just mentioned, and N-H ... pi hydrogen bonds. Finally, the CSD can suggest synthetic targets for medicinal chemistry; for example, molecules containing delocalised electron deficient groups such as trimethylammonium, pyridinium, thaizolium and dinitrophenyl have a good chance of binding to an active-site tryptophan. Although the CSD contains small-molecule crystal structures, not protein-ligand complexes, there is considerable evidence that the contacts seen in the two types of structures are similar. We have illustrated this a number of times in the present review and additional evidence has been given previously by Klebe. The major advantages of the CSD are its size, diversity and experimental accuracy. For these reasons, it is a useful tool for modellers engaged in rational inhibitor design. PMID- 9208368 TI - Reversible inhibition of enzymatic systems involving covalent intermediates. AB - Reversible inhibition phenomena are analyzed for enzymatic systems involving covalent intermediates, where the inhibitor can bind to the pure enzyme, the Henri-Michaelis complex or the covalent intermediate, or to two or three of these enzyme forms. Classical competitive, non-competitive or un-competitive phenomena can be observed in some cases but unexpected features are also observed. Complex phenomena sometimes prevail where increased kcat/Km values can be accompanied by decreased or increased kcat values. PMID- 9208369 TI - Inhibition of L-fucokinase from rat liver by L-fucose analogues in vitro. AB - By investigating the effects of more than 15 different L-fucose analogues on the activity of L-fucokinase (EC 2.7.1.52) from rat liver in vitro, certain structural requirements for potent inhibition of this enzyme were established. Of the novel compounds, 4,6-dideoxy-L-xylo-hexopyranose (4) and methyl 4,6-dideoxy-4 iodo-L-glucopyranose (9) were found to be competitive inhibitors with Ki-values of 0.5 mM and 5.0 mM respectively. Thus 4,6-dideoxy-L-xylo-hexopyranose is a better inhibitor of L-fucokinase than methyl-alpha-L-fucoside (1). Uptake of L fucose into rat hepatoma cells is reduced by 52% in the presence of the deoxy derivative (4), leading to a decrease of 45% in the incorporation of L-fucose into total cellular glycoproteins. PMID- 9208370 TI - Thermodynamic investigation of camel retina acetylcholinesterase inhibition by cyclophosphamide. AB - The present work addresses the estimation of energy parameters such as Gibb's free energy change (delta G), enthalpy change (delta H); entropy change (delta S) and activation energy (E a) of acetylcholinesterase (AChE, EC 3.1.1.7) from camel retina in the absence and presence of the antineoplastic drug cyclophosphamide (CP). A spectrophotometric method was used for the determination of AChE activity, which was the basis for determination of these parameters. The PZ factor (number of sterically and energetically favorable collisions occurring between CP and AChE) have also been studied in this investigation. The energy parameters obtained in the present investigation were compared with the values reported elsewhere. PMID- 9208371 TI - Inhibitory effect of drugs with a ketone group on reduction of acetohexamide catalyzed by carbonyl reductase from rabbit kidney. AB - The reduction of acetohexamide catalyzed by carbonyl reductase from rabbit kidney was inhibited by befunolol, moperone, levobunolol, daunorubicin and loxoprofen, which have a ketone group within their chemical structures and are substrates for the enzyme. A significant correlation was observed between the common logarithm of Vmax/Km values of the enzyme for befunolol, moperone, levobunolol and daunorubicin and the percentage inhibition of the enzyme, confirming that these drugs are competitive substrates of the enzyme with respect to acetohexamide. However, the plot for loxoprofen, a nonsteroidal anti-inflammatory drug with a ketone group, was apparently distant from the regression line obtained. Although nonsteroidal anti-inflammatory drugs with a ketone group such as suprofen and fenbufen were not reduced by the enzyme, they strongly inhibited the reduction of acetohexamide catalyzed by the enzyme. PMID- 9208372 TI - Inhibitory kinetic studies on rat hepatic and renal succinate dehydrogenase with guanidine. AB - Guanidine-induced alterations in substrate dependent kinetics of hepatic and renal succinate dehydrogenase (SDH) have been investigated under in vitro conditions. Guanidine hydrochloride (GuHCl) induced a mixed type of inhibition by decreasing the maximal velocity (Vmax) and increasing the Michaelis-Menten constant (Km). The competitive (Ki) and non-competitive (Ki) inhibitory constants were calculated. The values showed that the inhibitory influence of GuHCl is more due to decreased enzyme substrate affinity rather than reduction in the active site density of the enzyme as revealed by low Ki values. PMID- 9208373 TI - The anatomical relationships of the prefrontal cortex with limbic structures and the basal ganglia. AB - This paper briefly discusses the anatomical criteria that have been used to delineate the prefrontal cortex (PFC) from the (pre)motor cortical areas in the frontal lobe. Single anatomical criteria, such as cytoarchitecture, connectivity with the mediodorsal thalamic nucleus or a dopaminergic innervation, are insufficient to unequivocally define the PFC. It is argued that, with respect to a number of structural aspects, the prefrontal and the (pre)motor cortical areas must be viewed as a continuum, whereas a (functional) differentiation is based on the type of information that is being processed in different parts of the frontal lobe. The involvement of the PFC, like the premotor cortex, in a number of basal ganglia-thalamocortical circuits may be interpreted in the same way. The paper also summarizes the organization of the inputs from midline/intralaminar thalamic nuclei, the basal amygdaloid complex and the hippocampus into the PFC-ventral striatal system. The results of tracing studies in rats indicate that these thalamic and limbic inputs both at the level of the PFC and the ventral striatum show various patterns of convergence and segregation. This leads to the conclusion that the PFC-ventral striatal system consists of a number of smaller modules. PMID- 9208374 TI - Frontal-subcortical circuits: the anatomic basis of executive, social and motivated behaviors. AB - A series of discrete, parallel frontal-subcortical circuits have been demonstrated to link specific areas of the frontal lobe to areas within the basal ganglia and thalamus. A variety of circuit-specific behaviors can be described involving the dorsolateral prefrontal, orbitofrontal and anterior cingulate circuits. Interruptions or imbalance occurring at various levels within these closed looped circuits is felt to underlie the characteristic behavioral patterns seen. The intricate neurochemical arrangement of the striatum and the complex neurotransmitter interactions that occur within these key subcortical structures from the basis for modulatory influences that can affect these circuits. PMID- 9208375 TI - Neuropsychological and neuroimaging evidence for the involvement of the frontal lobes in depression. AB - The onset and reversibility of major depression is likely to be explained by diffuse neuromodulatory mechanisms rather than permanent abnormalities of connectivity and neurotransmission. However, the expression of mood state appears to involve fronto-striatal mechanisms. Lesions of the ventral frontal cortex give rise to profound modification of affect and behaviour not explained by effects on current intellectual function. These may represent the most extreme possible disturbances of emotional experience. Neuropsychological testing in major depression shows evidence of slowing in motor and cognitive domains with additional prominent effects on mnemonic function most marked in the elderly. Structural imaging with X-ray computed tomography or magnetic resonance imaging in older patients with major depression shows evidence of structural abnormality compared with controls. These findings are not highly localizing but they tend to confirm the role of cognitive impairment as an important age-related risk factor for major depression. Perfusion or metabolic imaging reflects both reversible changes in function and permanent loss of active neurones. The usual finding has been reductions in anterior brain structures in major depression. Hypoperfusion tends to be greatest in frontal, temporal and parietal areas and most extensive in older (male) patients; high Hamilton scores tend to be associated with reduced uptake. There have also been correlations in the cingulate cortex between increased perfusion and other aspects of the mental state. In general, reductions in frontal areas may be more likely in patients with impoverished mental states. The more prominent impairments of memory are likely to be associated with the finding of impaired temporal function or with a more diffuse failure of neuromodulation. PMID- 9208376 TI - Dopamine, the prefrontal cortex and schizophrenia. AB - Dysfunction of the prefrontal cortex (PFC) in schizophrenia has been suspected based on observations from clinical, neuropsychological and neuroimaging studies. Since the PFC receives a dense dopaminergic innervation, abnormalities of the mesocortical dopamine system have been proposed to contribute to the pathophysiology of schizophrenia. In this review, aspects of the anatomy, physiology and pharmacology of the mesencephalic-frontal cortical dopamine system as they may relate to schizophrenia are described, and evidence for altered dopaminergic neurotransmission in the frontal cortex of schizophrenic patients is presented. PMID- 9208377 TI - The nature of interactions involving prefrontal and striatal dopamine systems. AB - A number of converging lines of evidence from work in rodents suggest that dopamine (DA) function in the prefrontal cortex (PFC) and striatal terminal fields may be linked, possibly in an 'inverse' manner, whereby a change in prefrontal dopamine transmission in one direction occasions an opposite change in dopamine function in striatal territories. The present article considers the possible functional importance of this concept in the light of recent neuroanatomical data and new data from our own laboratory indicating that, at the neurochemical level, the basic finding of an inverse relationship between dopamine function in prefrontal and striatal regions also holds good in the non human primate. The main conclusion is that the simple idea of an inverse relationship between prefrontal and striatal dopamine systems emphasizing presynaptic release mechanisms is unlikely to underlie, solely, the full repertoire of functional interactions. Whilst there is evidence consistent with dynamic interactions between prefrontal and striatal dopamine release under some circumstances, specifically, during the early phases of aversive learning, a complete account of possible interactions between prefrontal and striatal dopamine systems requires consideration of additional factors. Such factors include: (1) the precise nature of the psychological function investigated, (2) the possibility of acute, localized changes in striatal postsynaptic function secondary to changes in presynaptic function and (3) the possibility of manipulations of prefrontal cortex leading to adaptive changes in striatal function, at a diffuse, neural systems level. PMID- 9208378 TI - Catecholamine regulation of the prefrontal cortex. AB - The catecholamines dopamine (DA) and norepinephrine provide an essential modulatory influence on the working memory and attentional functions of the prefrontal cortex (PFC). The following critique reviews evidence that (1) either insufficient or excessive DA D1 receptor stimulation is detrimental to PFC function, while DA stimulation of the D2 family of receptors may contribute to detrimental actions in PFC and (2) that norepinephrine has an important beneficial influence on PFC function through its actions at post-synaptic, alpha 2A adrenergic receptors, but impairs PFC function through actions at alpha 1 adrenergic receptors. Critical levels of catecholamine stimulation may be needed to optimize PFC cognitive function; high levels of catecholamine release during stress may serve to take the PFC 'off-line' to allow faster, more habitual responses mediated by the posterior and/or subcortical structures to regulate behavior. These studies have relevance to our understanding and treatment of disorders with prominent symptoms of PFC dysfunction. PMID- 9208379 TI - Noradrenergic mechanisms in the prefrontal cortex. AB - There is growing evidence that noradrenergic inputs to the prefrontal cortex (PFC) play an important role in regulating its function. This paper reviews the pharmacological control of noradrenaline (NA) release in this region, with particular reference to our studies using brain microdialysis, and also describes how NA levels are modulated by antidepressant and antipsychotic drugs. The suggestion that atypical antipsychotics such as clozapine and risperidone may produce clinical benefits by their ability to increase NA release is discussed. Finally, a new class of drugs, which show selectivity for imidazoline receptors is described. These compounds are shown to similarly increase extracellular NA in the PFC. Their potential utility as clinical treatments is discussed. PMID- 9208380 TI - Effects of ibotenic acid-induced loss of neurons in the medial prefrontal cortex of rats on behavioral vigilance: evidence for executive dysfunction. AB - Rats were trained in a previously validated task for the assessment of sustained attention, or vigilance. This task required the animals to discriminate between signals of variable lengths and non-signal events by making an appropriate lever press. The performance of sham-lesioned animals in this task was characterized by a signal-length dependent number of hits. Also, approximately 70 percent of the non-signals were correctly rejected. Ibotenic acid-induced lesions of the medial prefrontal cortex decreased the relative number of hits and correct rejections and, in essence, resulted in random lever selection. The lesion did not affect the number of omissions or side bias. Furthermore, the performance of lesioned animals was insensitive to the detrimental effects of distractors. The effects of the lesions do not support an interpretation in terms of sustained attention. Rather, the pattern of the lesioned animals' performance is speculated to reveal a fundamental disruption of decisional processes, reminiscent of the executive dysfunctions observed in patients with damage to ventromedial prefrontal areas or with schizophrenia. PMID- 9208381 TI - The frontal lobes and neurosurgery for psychiatric disorders. AB - The frontal lobe has been the main target for surgical treatment of mental illness over the last 60 years. Initially the surgery was crude and performed on patients with many different psychiatric disorders. Contemporary surgery utilizes stereotactic lesions which interrupt fronto-thalamic and/or fronto-cingulate fibres. The findings of clinical, neurochemical, neuroimaging, neuropsychological and physiological research in this area are summarized. Current advances in clinical neuroscience methods should be used in patients with these lesions to elucidate the neural substrate of post-operative changes and optimize clinical practice. PMID- 9208382 TI - Perspectives on the role of serotonergic mechanisms in the pharmacology of schizophrenia. PMID- 9208383 TI - Dangerous interaction with nefazodone added to fluoxetine, desipramine, venlafaxine, valproate and clonazepam combination therapy. PMID- 9208384 TI - Hypericum, an over the counter antidepressant? PMID- 9208386 TI - Fatty acid composition of lymphocytes and macrophages from rats fed fiber-rich diets: a comparison between oat bran- and wheat bran-enriched diets. AB - The effect of oat bran- (OBD) and wheat bran-enriched diets (WBD) on fatty acid composition of neutral lipids and phospholipids of rat lymphocytes and macrophages was investigated. In neutral lipids of lymphocytes, OBD reduced the proportion of palmitoleic acid (48%), whereas WBD reduced by 43% palmitoleic acid and raised oleic (18%), linoleic (52%), and arachidonic (2.5-fold) acids. In neutral lipids of macrophages, OBD increased palmitic (16%) and linoleic (29%) acids and slightly decreased oleic acid (15%). The effect of WBD, however, was more pronounced: It reduced myristic (60%), stearic (24%) and arachidonic (63%) acids, and it raised palmitic (30%) and linoleic (2.3-fold) acids. Neither OBD nor WBD modified the composition of fatty acids in phospholipids of lymphocytes. In contrast, both diets had a marked effect on composition of fatty acids in macrophage phospholipids. OBD raised the proportion of myristic (42%) and linoleic (2.4-fold) acids and decreased that of lauric (31%), palmitoleic (43%), and arachidonic (29%) acids. WBD increased palmitic (18%) and stearic (23%) acids and lowered palmitoleic (35%) and arachidonic (78%) acids. Of both cells, macrophages were more responsive to the effect of the fiber-rich diets on fatty acid composition of phospholipids. The high turnover of fatty acids in macrophage membranes may explain the differences between both cells. The modifications observed due to the effects of both diets were similar in few cases: an increase in palmitic and linoleic acids of total neutral lipids occurred and a decrease in palmitoleic and arachidonic acids of phospholipid. Therefore, the mechanism involved in the effect of both diets might be different. PMID- 9208387 TI - Dietary effect of a symmetrical triacylglycerol, 1,3-biseicosapentaenoyl-2-gamma linolenoyl glycerol, on fatty acid composition of guinea pigs. AB - The dietary effect of 1,3-biseicosapentaenoyl-2-gamma-linolenoyl glycerol (STG) on the fatty acid composition of guinea pigs was examined and compared with that of an eicosapentaenoic acid ethyl ester (EPA-E) and of a soybean oil (SBO) diet. In terms of content of plasma lipid, EPA-E had a greater hypolipidemic effect than STG. On the other hand, in terms of EPA incorporation, contents of EPA in liver lipid were almost the same in the STG and EPA-E groups. Considering that the amount of EPA administered in the EPA-E group was almost 1.5 times that of the STG group, EPA may be absorbed more effectively as the glycerol ester than as the ethyl ester in guinea pigs. In all the tissue lipids, the STG group had a higher unsaturation index (UI) than the EPA-E group even though there is a lower UI in the STG diet than the EPA-E diet. These results suggest that greater amounts of desaturase products as a whole were synthesized in the STG group than in the other two groups. The dihomo-gamma-linolenic acid/arachidonic acid (DGLA/AA) ratio in plasma total lipids in the STG group was 3.5 times that of SBO group, and the DGLA/AA ratio in the EPA-E group was half that of the SBO group. In liver lipid, the ratios of DGLA/AA and EPA/AA in the STG group were 0.687 and 0.488 (phosphatidylcholine fraction) and 0.237 and 0.752 (phosphatidylethanolamine fraction), respectively. The ratio of DGLA/AA as well as the high EPA/AA ratio obtained in the present study with the STG diet may lead to physiological alterations, including enhanced synthesis of 1- and 3-series eicosanoids. PMID- 9208385 TI - Expression of fatty acyl-CoA binding proteins in colon cells: response to butyrate and transformation. AB - Fatty acyl-CoA affect many cellular functions as well as serving as cellular building blocks. Several families of cytosolic fatty acyl-CoA binding proteins may modulate the activities of fatty acyl-CoA. Intestinal enterocytes contain at least three unique families of cytosolic proteins that bind fatty acyl-CoA: acyl CoA binding protein (ACBP), fatty acid binding proteins (including the liver, L FABP and intestinal, I-FABP), and sterol carrier protein-2 (SCP-2). Immortalized rat colon epithelial cell lines expressed only ACBP and SCP-2 at levels of 0.75 +/- 0.13 and 0.42 +/- 0.02 ng/microgram protein. Ras and src transformation increased colon cell density and differentially altered ACBP and SCP-2 expression without affecting I-FABP or L-FABP levels. ACBP levels were 1.8-fold and 1.5-fold increased in ras- and src-transformed cells, respectively. In contrast, SCP-2 expression was significantly decreased 55 and 67% in ras- and src-transformed cells, respectively. Butyrate treatment of ras- and src-transformed cells decreased cell proliferation up to 60-85% as compared to 25-30% in control cells. Butyrate treatment decreased ACBP expression in all cell lines but had no effect on the levels of SCP-2, I-FABP, or L-FABP. These studies suggest that the differential expression of ACBP and SCP-2 in rat colonic cell lines, as well as their modulation by butyrate, may be altered by cell transformation. PMID- 9208388 TI - Distribution of cholesterol sulfate and its anabolic and catabolic enzymes in various rabbit tissues. AB - Cholesterol sulfate (CS) recently has been shown to be involved in signal transduction pathway. To evaluate its functional significance, we determined the concentration of CS, and the specific activities of cholesterol sulfotransferase and CS sulfatase in various tissues of rabbit, and compared them with the concentration of sulfoglycolipids in rabbit tissues. CS was present in the epithelia and mucosa, but not in the tunica muscularis, of the digestive tract, trachea, uterine endometrium and uterine cervix. It was also present in lung, spleen, kidney, prostate, skin, hair, and nail at relatively high concentrations. Its concentration in the uterine endometrium was nine times higher in pseudopregnant rabbits than in nonpregnant rabbits because of activation of cholesterol sulfotransferase and inhibition of CS sulfatase in the pseudopregnant rabbits. Sulfoglycolipids were not detected in the uterine endometria of either non-pregnant- or pseudopregnant rabbits. However, sulfoglycolipids were detected at relatively high concentrations in the cerebrum, cerebellum, stomach, duodenum, jejunum, testis, and kidney of rabbits and thus the tissues in which both sulfolipids were detected were the gastrointestinal tract and kidney. In the digestive tract, the concentration of CS decreased in the order esophagus, stomach, duodenum, and jejunum, but that of sulfatide increased in the same order, indicating distribution of CS in the squamous epithelium. In addition, both CS and sulfatide were detected in the serum. On the other hand, CS sulfatase activity was detected in all tissues examined, even in hair, from which the enzyme was liberated by brief sonication, and its highest specific activity was detected in the liver. The specific activity of cholesterol sulfotransferase varied among the tissues examined and was found to be significantly high in the esophageal epithelium and the uterine endometrium of pseudopregnant rabbit, indicating involvement of cholesterol sulfation in the formation of epithelium. PMID- 9208389 TI - Sesamol as an inhibitor of growth and lipid metabolism in Mucor circinelloides via its action on malic enzyme. AB - Sesamol, a nonoil component of sesame seed oil, inhibited growth, fatty acid synthesis, and desaturation by Mucor circinelloides in vivo. Although sesamol also inhibited the growth of other fungi and yeasts, its effect on the lipid metabolism of M. circinelloides was exceptional. An enzymological study demonstrated that sesamol affected lipid synthesis primarily by the inhibition of malic enzyme activity, thereby limiting the NADPH supply for fatty acid synthesis and desaturation. Sesamol itself had no inhibitory effect on malic enzyme activity in vitro. A metabolite of sesamol is therefore probably responsible for the in vivo effects of sesamol on lipid metabolism. PMID- 9208390 TI - Modification of odd-chain length unsaturated fatty acids by hepatocytes of rainbow trout (Oncorhynchus mykiss) fed diets containing fish oil or olive oil. AB - Hepatocytes isolated from rainbow trout fed on diets containing either fish oil or olive oil were incubated with individual odd-chain length unsaturated fatty acids (19:1n-9, 19:2n-6, 19:3n-3, 21:2n-6, 21:3n-6, 21:4n-6, 21:3n-3, and 21:5n 3) to examine whether these fatty acids were substrates for modification by desaturation and elongation. All odd-chain length fatty acids were readily assimilated into the lipids of hepatocytes from both dietary groups of fish, but their conversion to longer-chain, more unsaturated derivatives was more pronounced with cells from trout fed olive oil. Thus, the conversion of 19:2n-6 and 21:2n-6 to 21:3n-6 and 21:4n-6, and of 19:3n-3 to 21:4n-3 and 21:5n-3, was most obvious in cells from the olive oil group, as was the conversion of 21:3n-6 and 21:3n-3 to 21:4n-6 and 21:4n-3, respectively. Elongation of 19:1n-9 to 21:1n 9 and 23:1n-9 occurred in cells from both groups. No 23:6n-3 was detectable as a product of 19:3n-3 or 21:3n-3. However, this fatty acid was a major product formed by cells from fish fed olive oil presented with 21:5n-3. Cells from both groups of fish incorporated 21:4n-6 and 21:5n-3 into their lipids largely without modification but chain-shortened around 40, 23, and 19% of the incorporated 21:2n 6, 21:3n-3, and 19:1n-9, respectively. The results demonstrate that odd-chain length unsaturated fatty acids can act as substrates for the desaturation, elongation, and chain-shortening systems of trout hepatocytes. PMID- 9208391 TI - Lipid content and fatty acid composition of 11 species of Queensland (Australia) fish. AB - The fatty acid composition of 11 species of fish caught of the northeast coast of Australia was determined. No fatty acid profiles have been previously published for fish from this area nor for nine of these species. Although the percentage of polyunsaturated fatty acid (PUFA) was the same as the calculated average for Australian fish (42.3%), the percentage of n-3 fatty acids was lower (24.4 +/- 5.4% vs. 30.7 +/- 10.1%) and the n-6 fatty acids higher (16.5 +/- 4.5% vs. 11.2 +/- 5.9%), P < 0.001 in each case. The major n-3 PUFA were docosahexaenoic (15.6 +/- 6.3%) and eicosapentaenoic acid (4.3 +/- 1.1%) while the major n-6 PUFA were arachidonic (8.3 +/- 3.2%) and n-6 docosatetraenoic acid (3.1 +/- 1.3%). The second-most abundant class of fatty acid was the saturates (31.6 +/- 3.5%) while the monounsaturates accounted for 17.4 +/- 4.3% of the total fatty acids. The monounsaturate with the highest concentration was octadecenoic acid (11.8 +/- 2.6%). There was a positive correlation between the total lipid content and saturated and monounsaturated fatty acids (r = 0.675 and 0.567, respectively) and a negative correlation between the total lipid content and PUFA (r = 0.774). PMID- 9208392 TI - Plasma kinetic behavior in hyperlipidemic subjects of a lipidic microemulsion that binds to low density lipoprotein receptors. AB - It was previously reported that a protein-free microemulsion (LDE) with structure roughly resembling that of the lipid portion of low density lipoprotein (LDL) was presumably taken up by LDL receptors when injected into the bloodstream. In contact with plasma, LDE acquires apolipoproteins (apo) including apo E that would be the ligand for receptor binding. Currently, apo were associated to LDE by incubation with high density lipoprotein (HDL). LDE-apo uptake by mononuclear cells showed a saturation kinetics, with an apparent K(m) of 13.1 ng protein/mL. LDE-apo is able to displace LDL uptake by mononuclear cells with a Ki of 11.5 ng protein/mL. LDE without apo is, however, unable to displace LDL. The uptake of 14C-HDL is not dislocated by increasing amounts of LDE-apo, indicating that HDL and LDE-apo do not bind to the same receptor sites. In human hyperlipidemias, LDE labeled with 14C-cholesteryl ester behaved kinetically as expected for native LDL. LDE plasma disappearance curve obtained from eight hypercholesterolemic patients was markedly slower than that from 10 control normolipidemic subjects [fractional clearance rate (FCR) = 0.02 +/- 0.01 and 0.12 +/- 0.04 h-1, respectively; P < 0.0001]. On the other hand, in four severely hypertriglyceridemic patients, LDE FCR was not significantly different from the controls (0.07 +/- 0.03 h-1). These results suggest that LDE can be a useful device to study lipoprotein metabolism. PMID- 9208394 TI - Noninvasive characterization of neonatal adipose tissue by 13C magnetic resonance spectroscopy. AB - In vivo 13C magnetic resonance spectroscopy (MRS) was applied noninvasively to analyze the fatty acid composition of adipose tissue in 21 full-term newborn infants and 6 mothers. In order to assess the effects of gestational and postnatal age on adipose tissue composition, we studied preterm infants at birth, term infants at the ages of 6 wk and at 6 mon. We also investigated the influence of maternal diet on infant adipose tissue composition by studying the breast-fed infants of women who maintained either an omnivore or a vegan diet. Significant differences were observed in adipose tissue composition of neonates compared with their mothers. Neonates had more saturated and less unsaturated fatty acids than their mothers (P < 0.01). We also observed changes in adipose tissue composition with maturity. From birth to 6 wk of age 13C MR spectra showed a significant increase in the amount of unsaturated fatty acids, particularly polyunsaturated fatty acids (P < 0.01). Similarly, differences were seen as a result of gestational age. Preterm infants had relatively fewer unsaturated fatty acids than full-term infants. A greater proportion of these unsaturated fatty acids were polyunsaturated. Our results demonstrate that 13C MRS can be utilized to assess noninvasively neonatal adipose tissue lipid composition and to monitor the effects of developmental changes due to gestational age and oral feeding. PMID- 9208393 TI - The effect of short-term diets rich in fish, red meat, or white meat on thromboxane and prostacyclin synthesis in humans. AB - Foods which increase tissue arachidonic acid levels have been proposed to increase thrombosis tendency, presumably through increased platelet aggregation. This study examined the effect of doubling the dietary arachidonic acid (20:4n-6) using meat- or fish-based diets on the systemic production of prostacyclin (PGI2) and thromboxane (TXA2) in 29 healthy, nonsmoking adults. There were three, 3-wk low-fat dietary periods (< 15% energy as fat) in which subjects consumed a vegetarian diet for 1 wk followed by 2 wk on diets containing meat or fish as sources of 20:4n-6. Between each diet period, there was a 3-wk washout period, during which subjects returned to their normal diets. The level of 20:4n-6 consumed during the last 2 wk of each study was approximately double the usual intake (mean 140 mg/d), while the mean eicosapentaenoic acid (20:5n-3) content of the diets varied from 1 mg/d on the white meat diet to 70 mg/d on the red meat diet and to 847 mg/d on the fish diet. The serum phospholipid (PL) 20:4n-6/20:5n 3 ratios were 11:1 on the vegetarian diet, 15:1 on the white meat diet, 8:1 on the red meat diet, and 2:1 on the fish diet (P < 0.001). Neither white nor red meat diets affected platelet 20:4n-6 levels, platelet aggregation, ex vivo platelet TXB2 production, or the systemic PGI2 or TXA2 production as measured by gas chromatography-mass spectrometry analysis of the excretion levels of the principal urinary metabolites 2,3-dinor-6-keto-PGF1 alpha (PGI2-M) and 11-dehydro TXB2 (TXA2-M), respectively. The fish diet decreased the 20:4n-6/20:5n-3 ratio in platelet PL from the baseline level of 45:1 to 13:1 (P < 0.001), had no effects on platelet aggregation, but significantly decreased platelet TXB2 production (collagen-stimulated) and TXA2-M production, while PGI2-M levels were unaltered. These results indicate that short-term diets which double the usual 20:4n-6 intake using white meat (175-330 g/d) or red meat (275-530 g/d) are not associated with an increased TXA2 production, but this does not rule out the adverse effects of 20:4n-6 at higher levels in the diet, or for more prolonged periods. Short-term diets containing fish (100-200 g/d with 90-210 mg/d 20:4n-6 and approximately 650-1000 mg/d 20:5n-3) led to significant increases in platelet 20:5n-3 levels and a decrease in the ex vivo and systemic TXA2 production. PMID- 9208395 TI - Thermotropic phase behavior of in vivo extracted human stratum corneum lipids. AB - The thermotropic phase behavior of lipids extracted either in vivo from inner forearm (SCLE) or plantar callus (PC) was investigated by differential scanning calorimetry and small angle X-ray diffraction. PC composition was chromatographically modified (MPC) by eliminating the more polar lipids in order to evaluate their role. Analysis of composition confirms the potential use of PC as a source of stratum corneum lipids. MPC and SCLE exhibit similar differential scanning calorimetry (DSC) profiles with a main transition around 50 degrees C attributed to the solid-to-liquid phase transition of the ceramides. The absence of a transition around 50 degrees C for PC suggests the possible perturbation of ceramide packing by the significantly high proportion of phospholipids. X-ray data suggest a high miscibility of sebum components in stratum corneum lipids with possible modification of chain packing. The MPC patterns show a lipid phase separation which underscores the role of polar lipids in cholesterol/free fatty acids/sterol esters/ceramides structural cohesion. PMID- 9208396 TI - The relationship between the structure of monoalkyl branched saturated triacylglycerols and some physical properties. AB - Three different physical properties, the gel point ("solidification point"), the refractive index and the density, were determined and related to the structure of the branched triacylglycerols. A four-factor central composite face-centered design was constructed where the four variables were the length of the main chain, the branching position, the length of the side chain, and the number of branched fatty acyl groups attached to the glycerol backbone. Second-order models were calculated in which the three physical properties were related to the structure. Four additional branched triacylglycerols were analyzed in order to confirm the validity of each model. Contour plots are shown in order to visualize the prediction equations which were obtained. PMID- 9208397 TI - On the syntheses of triacylglycerols from branched saturated fatty acids. AB - A number of triacylglycerols with branched acyl groups were prepared via 1,2 isopropylidene glycerol for the purpose of studying three different physical properties: gel point, refractive index, and density. The monoacid triacylglycerols were prepared either via the corresponding acids or the acyl chlorides. PMID- 9208398 TI - A new lipid from an Australian marine sponge, Callyspongia sp. AB - A specimen of the sponge Callyspongia sp. collected off the coast of New South Wales, Australia, has yielded the novel lipid (6Z, 9Z, 12Z, 15Z)-1,6,9,12,15 octadecapenten-3-one, together with (4Z, 7Z, 10Z, 13Z)-4,7,10,13 hexadecatetraenoic acid. PMID- 9208399 TI - Gas-liquid chromatographic properties of positional isomers of methyl thia, selena, and tellura laurate analogs. AB - Gas-liquid chromatographic analyses of three complete series of synthetic positional isomers of methyl thia, selena, and tellura laurate analogs were carried on a nonpolar (SE-30) and a polar (SP-2330) stationary phase. The average ECL (equivalent chain length) values of the thia, selena, and tellura laurate on SE-30 stationary phase were 13.8, 14.8, and 15.7, respectively, while on SP-2330 the average values for the same series were 17.1, 19.0, and 19.1, respectively. Positional isomers with the heteroatom at the 2-position exhibited the lowest ECL values, while those with the heteroatom at the omega-1 position gave the highest ECL values and were readily separated from the other positional isomers of the same series of analogs by this technique. PMID- 9208400 TI - Gaseous transmitters and neuroendocrine regulation. AB - Recent work has demonstrated that the brain has the capacity to synthesize impressive amounts of the gases nitric oxide (NO) and carbon monoxide (CO). There is growing evidence that these gaseous molecules function as novel neural messengers in the brain. This article reviews the pertinent literature concerning the putative role of NO and CO as critical neurotransmitters and biological mediators of the neuroendocrine axis. Abundant evidence is presented which suggests that NO has an important role in the control of reproduction due to its ability to control GnRH secretion from the hypothalamus. NO potently stimulates GnRH secretion and also appears to mediate the action of one of the major transmitters controlling GnRH secretion, glutamate. Evidence is presented which suggests that NO stimulates GnRH release due to its ability to modulate the heme containing enzyme, guanylate cyclase, which leads to enhanced production of the second messenger molecule, cGMP. A physiological role for NO in the preovulatory LH surge was also evidenced by findings that inhibitors and antisense oligonucleotides to nitric oxide synthase (NOS) attenuate the steroid-induced and preovulatory LH surge. CO may also play a role in stimulating GnRH secretion as heme molecules stimulate GnRH release in vitro, an effect which requires heme oxygenase activity and is blocked by the gaseous scavenger molecule, hemoglobin. Evidence is also reviewed which suggests that NO acts to restrain the hypothalamic-pituitary-adrenal (HPA) axis, as it inhibits HPA stimulation by various stimulants such as interleukin-1 beta, vasopressin, and inflammation. This effect fits a proinflammatory role of NO as it leads to suppression of the release of the anti-inflammatory corticosteroids from the adrenal. Although not as intensely studied as NO, CO has been shown to suppress stimulated CRH release and may also function to restrain the HPA axis. Evidence implicating NO in the control of prolactin and growth hormone secretion is also reviewed and discussed, as is the possible role of NO acting directly at the anterior pituitary. Taken as a whole, the current data suggest that the diffusible gases, NO and CO, act as novel transmitters in the neuroendocrine axis and mediate a variety of important neuroendocrine functions. PMID- 9208401 TI - Chicken GnRH II-like peptides and a GnRH receptor selective for chicken GnRH II in amphibian sympathetic ganglia. AB - Amphibia, like most vertebrate species, have two forms of GnRH, namely [Arg8]GnRH (mammalian GnRH) and [His5,Trp7,Tyr8] GnRH (chicken GnRH II). The differential distribution of the two peptides in the amphibian brain suggests that they may play different roles. Mammalian GnRH, which is found predominantly in the hypothalamus, is most likely the prime regulator of gonadotropin release, while chicken GnRH II, which occurs predominantly in the midbrain and hindbrain, may play a neuromodulatory role. In amphibian sympathetic ganglia, GnRH has been demonstrated to be a neurotransmitter where its release from the presynaptic nerve terminals reversibly inhibits M current, a time- and voltage-dependent potassium current. The occurrence of GnRH in sympathetic ganglia extracts from two amphibian species was investigated. Chicken GnRH II-like immunoreactivity was detected in extracts of bullfrog (Rana catesbeiana) and platanna (Xenopus laevis) sympathetic ganglia after high performance liquid chromatography. Under the chromatographic conditions used, a second unknown peptide co-eluted with synthetic mammalian GnRH, but showed no cross-reactivity with specific mammalian GnRH antisera. To test the possibility of the presence of a chicken GnRH II receptor in sympathetic ganglion neurones, competition binding of membranes extracted from the sympathetic ganglia of the two amphibian species was investigated with 125I-labelled GnRH agonists. The binding of 125-I-[His5,D Arg6,Trp7,Tyr8]GnRH (a chicken GnRH II agonist) to membranes from the sympathetic ganglia of both amphibian species was specific and had a higher affinity than chicken GnRH II, mammalian GnRH and a mammalian GnRH agonist [D-Ala6,NMe Leu7,Pro9-NHEt]GnRH. These findings suggest that endogenous chicken GnRH II may play a role in synaptic transmission in the sympathetic ganglia via a receptor specific for chicken GnRH II. PMID- 9208402 TI - Lesions of gonadotropin-releasing hormone-immunoreactive terminal nerve cells: effects on the reproductive behavior of male dwarf gouramis. AB - Functions of the terminal nerve (TN) are largely unknown. To examine whether gonadotropin-releasing hormone (GnRH)-immunoreactive TN cells (TN-GnRH cells) are involved in the control of reproductive behavior, effects of lesions of TN-GnRH cells were studied in male dwarf gouramis, Colisa lalia. After bilateral electrolytic lesion, a characteristic impairment was observed in one of the repertoires of male reproductive behavior, nest-building. The occurrence of mating trials in which males showed no nest-building was increased. However, the incidence of nest-building behavior during postoperative trials was not affected by the lesion. No impairment was observed in other reproductive repertoires. These results suggest that (1) TN-GnRH cells are involved in the control of the threshold for nest-building behavior initiation and (2) TN-GnRH cells are not a prerequisite for other aspects of reproductive behavior in the male gouramis. PMID- 9208403 TI - Physiologically induced Fos expression in the hypothalamo-hypophyseal system of Xenopus laevis. AB - The amphibian Xenopus laevis adjusts the color of its skin to the degree of background illumination. The neuroendocrine mechanism responsible for this adaptation behavior involves various brain centers that control the synthesis and release of alpha-melanophore-stimulating hormone (alpha-MSH) by the pituitary melanotrope cells. The aim of the present study was to investigate the possible use of Fos as a tool to determine the activity of known and novel components of this mechanism. For this purpose, a quantitative Fos-immunocytochemical method (ABC) was successfully introduced for Xenopus, and the degree of specificity of background illumination as a regulator of Fos expression was tested by comparing this stimulus with two other stimuli, viz. a strong stressor (saline immersion) and a mild stressor ('handling'). Without stimulation basal Fos-like immunoreactivity (Fos-LI) was found in the telencephalon, the lateral pallium, the anterior, central and lateral thalamic nuclei, the suprachiasmatic nucleus, the ventral hypothalamic nucleus and the torus semicircularis. Handling had no effect on this basal pattern of Fos-LI. Saline immersion induced Fos-LI only in the magnocellular preoptic nucleus, the corticotrope cells and, less strongly, in the melanotrope cells. Melanotropes, and no other cells, expressed Fos-LI very strongly when Xenopus was transferred from a white to a black background. This Fos-LI expression continued to increase up to 7 days of stimulation. When such toads were returned to a white background it took the same time before Fos-LI expression significantly dropped. It is concluded that the ABC-Fos immunocytochemistry can be successfully applied to assess the occurrence and degree of expression of Fos-LI in the Xenopus brain and pituitary gland. The prolonged expression of Fos-LI in the pars intermedia under black background stimulation and the presence of an AP-1 binding site on the Xenopus proopiomelanocortin gene suggest an important role for c-Fos and/or Fos-related antigens in the control of the biosynthesis and secretion of alpha-MSH by the Xenopus melanotrope cell. PMID- 9208404 TI - Dopamine receptors influence vasoactive intestinal peptide release from turkey hypothalamic explants. AB - Vasoactive intestinal peptide (VIP) is a significant prolactin-releasing factor (PRF) in avian species, and dopamine (DA) exhibits both a stimulatory and inhibitory influence upon this prolactin (PRL) secretion. The stimulatory effect of DA upon PRL release appears to be mediated by VIP. This study investigated DAergic actions upon VIP release using turkey hypothalamic explants perifused with DA and its agonists or antagonists. VIP release was stimulated by DA in a dose-dependent manner (10 nmol DA/min, from 67.2 +/- 3.9 to 164.3 +/- 3.1 pg/5 min; 100 nmol DA/min, from 70.1 +/- 3.2 to 291.0 +/- 7.5 pg/5 min; 1,000 nmol DA/min, from 72.0 +/- 4.8 to 501.0 +/- 24.7 pg/5 min). The D1 DA receptor antagonist (R+)-SCH-23390 HCl completely negated the stimulatory effect of DA (100 nmol/min) upon VIP release. Perifusion with the D2 DA receptor antagonist S( )-eticlopride HCl by itself stimulated VIP release from the hypothalamic explants, increasing VIP from 38.1 +/- 5.3 to 161.9 +/- 9.7 pg/5 min, where release stabilized until perifusion was terminated. The D1 DA agonist (+)-SKF 38393 HCl increased VIP release from 52.7 +/- 4.6 to 192.6 +/- 16.9 pg/5 min, and this stimulated release was partially inhibited by the D2 DA receptor agonist R( )-NPA HCl (from 192.6 +/- 16.9 to 139.7 +/- 13.8 pg/5 min). These results suggest that VIP secretion is in part regulated by possible opposite actions between stimulatory D1 and inhibitory D2 DA receptors in the turkey hypothalamus. PMID- 9208406 TI - Dopaminergic neurons in periventricular and arcuate nuclei of proestrous and ovariectomized rats: endogenous diurnal rhythm of Fos-related antigens expression. AB - The expression of Fos-related antigens (FRAs) in the A12 and the A14 hypothalamic dopaminergic neurons was compared throughout the day in proestrous (PRO) and ovariectomized (OVX) rats to establish the relationship between secretion of prolactin (PRL) and dopaminergic neuronal activation. Animals with intact ovaries were sacrificed at 11:00, 13:00, 15:00, 17:00 and 21:00 h on the day of proestrus and 06:00 and 09:00 h in the morning of estrus. OVX animals were sacrificed at the same time points on the 12th day after surgery. Double-label immunocytochemistry was performed by using antibodies against FRAs as markers of tonic neuronal activity and tyrosine hydroxylase to identify dopaminergic neurons. Serum PRL levels were determined by radioimmunoassay. A pattern of FRAs expression was present in the A14 and A12 neurons of PRO rats. The incidence of FRAs expressing neurons was the highest in the first half of the day when the PRL levels were low, and decreased prior to the surge of serum PRL during the afternoon. This pattern was present in the dopaminergic neurons of the periventricular nucleus (A14), and in all portions of the arcuate nucleus (A12) except the ventrolateral portion of the middle arcuate nucleus. A similar pattern of FRAs expression existed in the A14 and A12 neurons of OVX rats in spite of that there were no detectable changes in serum PRL levels. However, the amplitude of decrease in the incidence of FRAs-labeled neurons was lower in OVX than in PRO rats. These data suggested that similar to the control of hypothalamic PRL releasing factors, the activation/deactivation pattern of the A12 and the A14 neurons is governed by a seemingly endogenous rhythmic input. The incidence of this rhythm is independent of the reproductive state, but its amplitude is enhanced by ovarian steroids. The disparity between the FRAs expression and serum PRL levels in OVX rats indicated that the effect of this endogenous dopaminergic rhythm upon PRL secretion is dependent on the ovarian steroid background. PMID- 9208407 TI - Role of the hypothalamic-pituitary-adrenal axis in the suppression of luteinizing hormone release by delta-9-tetrahydrocannabinol. AB - The ability of cannabinoids to affect anterior pituitary luteinizing hormone (LH) secretion has been largely attributed to a central nervous system site of action, however, the mechanism(s) by which cannabinoids alter LH release remains unclear. In the present study, the acute administration of delta-9-tetrahydrocannabinol (THC) produced a dose-related suppression of plasma LH and stimulation of adrenocorticotropin (ACTH) levels in ovariectomized female rats. To determine if activation of the hypothalamic-pituitary-adrenal axis was involved in the ability of THC to inhibit LH release, female rats were either pretreated with the corticotropin-releasing hormone (CRH) receptor antagonist, alpha-helical CRH, or were adrenalectomized prior to acute THC administration, in order to assess the roles of CRH and corticosterone in the ability of THC to suppress LH secretion. A low dose of THC (0.5 mg/kg b.w., iv) produced a decrease in plasma LH levels at 20 and 40 min posttreatment in ovariectomized, sham adrenalectomized rats. However, in adrenalectomized animals, plasma LH levels were suppressed at 40 min and remained decreased at 80 min following THC administration. Thus, the duration of LH suppression following THC treatment was significantly increased in adrenalectomized versus sham adrenalectomized rats (p < 0.05). Furthermore, pretreatment with the CRH receptor antagonist, alpha-helical CRH (100 micrograms/5 microliters, icv), 30 min before THC administration, attenuated the ability of a high THC dose (1.0 mg/kg) to inhibit LH release in ovariectomized rats. Together, these results demonstrate that THC has significant effects on LH and ACTH secretion in ovariectomized rats and suggest that THC-induced CRH activation, but not corticosterone release, plays a role in the suppression of LH release by cannabinoids. PMID- 9208408 TI - Infarcts in the vascular distribution of the middle cerebral artery in preterm and fullterm infants. AB - Twenty-three infants with an infarct in the territory of the middle cerebral artery are reported. The diagnosis was made using cranial ultrasound in all, confirmed on postmortem in two cases and on MRI, performed during the neonatal period or in infancy, in 18 of the 20 survivors. Involvement of the main branch was present in 7 cases and three of these had a gestational age of less than 35 weeks. In the other 16 infants, involvement of a cortical branch or one or more of the lenticulostriate branches was present and all but three of these had a gestational age of 34 weeks or less. While involvement of the main branch was usually diagnosed on postnatal day 1 or 2 using ultrasound, involvement of the lenticulostriate branches was noted as a wedgeshaped echogenic lesion in the caudate nucleus, thalamus or putamen, between day 4 up till day 24, and at term age in one of the cases. Neurodevelopmental outcome of those with involvement of the main branch was disappointing as all survivors developed a hemiplegia, associated with epilepsy in two; while so far only three of the other 16 infants developed cerebral palsy, one a hemiplegia and one athetoid cerebral palsy. Global delay was present in a further three cases. Infarcts in the region of the middle cerebral artery can occur in both preterm as well as fullterm infants. Involvement of the main branch also occurred in infants with a gestational age below 35 weeks and resulted in the development of a hemiplegia in all survivors. Involvement of one of the other branches was especially common in preterm infants, who had a more favourable outcome. As the lesion in the latter group was usually not present before the end of the first week, serial ultrasound up till term age is needed in order to identify these lesions. PMID- 9208409 TI - MRI assessment of myelination of motor and sensory pathways in the brain of preterm and term-born infants. AB - A study was performed in 48 neurologically normal preterm and term-born infants, with a postconceptional age at MRI varying between 30 2/7 and 46 weeks and a mean age of 34 2/7 weeks. The purpose of the study was to determine the normal progress of myelination on MRI in that age range. T1- and T2-weighted images of the brain were assessed for changes in signal intensity of white matter relative to gray matter. Multiple sites in the brainstem, cerebellum and cerebral hemispheres were assessed separately. The findings were correlated with the ages of the infants. As judged from relative signal intensities, myelin was present at the post-conceptional age of 30-34 weeks in the following structures: tegmentum pontis (in particular medial lemniscus), superior and inferior colliculi, decussation of the superior cerebellar peduncles, crura cerebri, ventrolateral thalamus, lateral globus pallidus, dorsolateral putamen, dentate nucleus, middle and superior cerebellar peduncles, vermis cerebelli, cortex bordering the central sulcus and hippocampus. Little progress in myelination was noticed up to the post conceptional age of 46 weeks. Between 34 and 46 weeks, myelin appeared in the lateral part of the posterior limb of the internal capsule and in the central part of the corona radiata and became more prominently visible in the cortex bordering the central sulcus. PMID- 9208410 TI - Dihydropyrimidinase deficiency, a progressive neurological disorder? AB - A case of a child presenting with congenital abnormalities at birth is reported. The early development remained severely retarded and acquired skills minimally. The head circumference centile decreased. Magnetic resonance imaging showed progressive neuronal atrophy and secondary delay in myelination. Dihydropyrimidine concentrations in body fluids were quantitated by NMR spectroscopy. Enzymatic assay in the liver biopsy revealed total deficiency of dihydropyrimidinase (DHP) (5,6-dihydropyrimidine amidohydrolase; EC 3.5.2.2). As such, the patient is the first with enzymatically proven DHP deficiency. Thus far dihydropyrimidinuria has been reported in three other patients with a variety of neurological abnormalities. A relation of the enzyme deficiency with the neurodegenerative clinical course in our patient is suggested. PMID- 9208411 TI - The effect of behavioural states on cerebral oxygenation during endotracheal suctioning of preterm babies. AB - Near infrared spectroscopy (NIRS) was used to investigate the effect of behavioural states on changes of oxygenated (O2Hb), deoxygenated haemoglobin (HHb) and total haemoglobin (tHb), during endotracheal suctioning. In an open prospective design, NIRS measurements have been done during 20 suctioning episodes in 13 preterm neonates. Heart rate, arterial oxygen saturation, and carbon dioxide tension were monitored continuously. Behavioural state (BS) observations were made and documented as well. The statistical analysis showed that in patients who were active, with crying periods during suctioning (behavioural states 4-5), changes of oxygenated (p < 0.005) and deoxygenated haemoglobin (p < 0.05), as well as of arterial oxygen saturation (p < 0.05) and heart rate (p < 0.05) were significantly greater than in patients who were quiet with predominant behavioural state 1, 2 and 3. These results underline the influence of behavioural states on the physiological answers to endotracheal suctioning. NIRS proved to be a valuable tool to evaluate possible harmful effects of different suctioning techniques. PMID- 9208414 TI - Acute near-fatal parainfectious cerebellar swelling with favourable outcome. AB - The clinical course, neuro-imaging features and neuropathologic findings in a child with para-infectious acute cerebellar swelling are described. Reversible transtentorial and transforaminal herniations occurred and required emergency posterior fossa decompression with external ventricular drainage. Neuropathologic examination of a cerebellar biopsy demonstrated a subacute pathogen-free cerebellitis. Following neurosurgical intervention and steroid therapy, symptoms and signs resolved and the patient is well 3 years later. Acute para-infectious cerebellar swelling is potentially fatal unless recognised and treated early in its evolution. PMID- 9208405 TI - Effects of ovarian steroid hormones on dopamine-controlled prolactin secretory responses in vitro. AB - Dopamine (DA), a well-established inhibitor of prolactin (PRL) secretion, has also been shown to stimulate PRL secretion from the anterior lobe of the pituitary gland of the rat. It has been reported that low doses of DA stimulate PRL whereas high doses inhibit PRL secretion. Our laboratory has previously reported that these PRL secretory responses are dependent upon the stages of the estrous cycle of the rat. The objective of the present study was to determine the steroid requirements for the differing PRL secretory responses to low and high doses of DA in perifusion. Animals were ovariectomized (OVX) and immediately given Silastic implants containing estradiol (E2) which has previously been shown in our laboratory to produce blood levels of 70-100 pg/ml, progesterone which has previously been shown in our laboratory to produce blood levels of 30-40 ng/ml, or the combination. OVX rats served as controls. Ten days later, the anterior lobes of the pituitary glands were harvested and enzymatically dissociated. Cells were mixed with Sephadex G-10 and placed in six 0.5-ml perifusion chambers (1 x 10(6) cells/chamber). Cells were perifused for 24 h with Dulbecco's modified Eagle's medium containing 0.2% bovine serum albumin and 0.1 mM ascorbic acid. The PRL secretory pattern was characterized in response to the following treatment sequence: (1) 30 min media alone (maximally uninhibited); (2) 24 min 100 pM DA; (3) 30 min media alone (DA withdrawal); (4) 24 min 1 microM DA, and (5) 30 min media alone (DA withdrawal). PRL secretion in the presence of 100 pM DA was unchanged in cells obtained from OVX animals, but exposure to 1 microM DA inhibited PRL release in these cells. Subsequent withdrawal stimulated PRL secretion relative to that of the initial exposure to media alone. The responses of cells from OVX rats implanted with progesterone alone was indistinguishable from those of the OVX controls. In cells obtained from animals implanted with E2 alone, 100 pM DA and its subsequent withdrawal neither stimulated nor inhibited PRL secretion. In contrast, 1 microM DA exposure initially stimulated and then inhibited PRL secretion in cells from E2-treated animals. Here, subsequent withdrawal of DA enhanced PRL secretion. In cells obtained from E2+progesterone treated animals, 100 pM DA exposure robustly enhanced PRL secretory responses. Withdrawal of this dose of DA had no further effect on PRL secretion. However, exposure of E2+progesterone-treated cells to 1 microM DA robustly stimulated and subsequently only slightly inhibited PRL secretion. The results of these studies suggest that inhibition of PRL secretion by DA is independent of ovarian steroids while E2 and progesterone in combination favors stimulation of PRL secretion in response to DA. Taken together, these data suggest that PRL secretory responses in this system are determined by the ovarian steroid milieu. PMID- 9208415 TI - Central pontine myelinolysis associated with acquired folate depletion. AB - After long-standing malnutrition a 15-month-old boy with signs of kwashiorkor was admitted in a moribund state with serious hyponatraemic dehydration, hypothermia, somnolence, and signs of a pontine disconnection syndrome. Folic acid levels were below the detection level in the presence of normal cobalamin levels. MRI of the brain showed global volume loss and signal abnormalities on the T2-weighted images suggestive for central pontine myelinolysis (CPM). Brainstem acoustic evoked responses have remained normal. The serious metabolic and nutritional derangements required substitution of folic acid, vitamins and trace elements as well as slow correction of hyponatraemic dehydration with return of the sodium level over a period of four days. This therapeutic regimen resulted in complete neurological recovery. Follow-up MRI documented normalisation of the initial pathologic findings. The hypothesis was put forward linking the pathogenesis of CPM with the combination of folate depletion and superimposed hyponatraemic dehydration. The previously acquired folate depletion could affect normal appositional function of myelin basic protein molecules due to insufficient methylation of arginine in position 107. The subsequent development of intramyelinic edema and CPM will then be triggered by the superimposed hyponatraemic dehydration. The verification of this hypothesis requires further investigations. PMID- 9208412 TI - Fetal akinesia sequence caused by nemaline myopathy. AB - Nine patients with the characteristic signs of fetal akinesia sequence (polyhydramnion, multiple joint contractures and lung hypoplasia) are described. In 8 of the 9 patients nemaline myopathy could be demonstrated with histology. The ninth patient presented the same phenotype as his 4 affected siblings in whom the nemaline myopathy could be histologically proven. Seven of the patients belonged to 2 families; the other 2 patients were isolated cases. In one fetal case nemaline myopathy was documented at week 22 of gestation. These observations demonstrate that nemaline myopathy can cause the fetal akinesia sequence, with onset of first symptoms as early as the beginning of the second trimester of pregnancy. PMID- 9208416 TI - An association between optic glioma and other tumours of the central nervous system in neurofibromatosis type 1. AB - Neurofibromatosis type 1 (NF1) has a very heterogeneous phenotype. It is not currently possible to predict which patients will have mild disease and which will develop serious complications. Medical management of patients with NF1 might be improved if subgroups of patients who are at especially high (or low) risk for particular complications could be identified. We have begun an analysis of NF1 patients in the National Neurofibromatosis Foundation International Database (NNFFID) to identify possible associations between the occurrence of clinical features. A striking association has been observed between the presence of optic glioma and of other central nervous system (CNS) tumours in NF1 patients. This association is not dependent on the effect of age. No association is seen between optic glioma and non-CNS neoplasms. The association of optic glioma and other intracranial neoplasms in patients with NF1 suggests that there are fundamental pathophysiological differences between patients with and without optic glioma. PMID- 9208413 TI - Prenatal ultrasound abnormalities in a patient with generalized neurofibromatosis type 1. AB - We describe a patient with an unusual neonatal disseminated form of neurofibromatosis (NF1). Prenatal ultrasound studies, at 35 weeks of gestation, revealed ambiguous external genitalia, an increased biparietal diameter and a decreased growth of long bones. Postnatal examination displayed generalized neurofibromatosis, with perineal, thoracic and spinal cord invasion by tumors. Spinal cord compression was responsible for paraparesis. The child died of a pulmonary infection at five years of age. No previous report of such prenatal abnormalities has been described. Genetic counselling is difficult because of the variable expression of the illness. PMID- 9208417 TI - Moyamoya disease in a child with glycogen storage disease type Ia. AB - A three-year-old child affected by glycogen storage disease (GSD) type Ia presented with acute hemiplegia secondary to Moyamoya disease. So far, the association of moyamoya with GSD Ia had only been reported twice. The rarity of both conditions makes their association unlikely to be a chance event and an etiological relationship between them must be considered. PMID- 9208418 TI - Landau-Kleffner syndrome and sulthiame. PMID- 9208419 TI - Molecular analysis of HTLV-I and HTLV-II isolates from Italian blood donors, intravenous drug users and prisoners. AB - Using molecular methods three or five major variants of HTLV-I have been identified; moreover two subtypes of HTLV-II defined as HTLV-IIa and HTLV-IIb with six variants within each of these groups have been described. In the present study we analysed proviral DNA obtained from the peripheral blood mononuclear cells (PBMCs) of a significant group of Italian intravenous drug users (IVDUs), prison inmates and blood donors (BDs) who were HTLV antibody positive. Restriction fragment length polymorphism (RFLP) of amplified LTR region with ApaI, NdeI, DraI, SacI and MaeIII endonucleases was used to define the HTLV-I subtypes, while the different variants of HTLV-IIa and -IIb were defined by RFLP of the LTR region with the AvaII, BglI, SauI, XhoI and BanII endonucleases. The four HTLV-I isolated from BDs were characterized as C type. All the 11 HTLV-II detected in the IVDUs were HTLV-IIb4, while among the prisoners one HTLV-IIb5 and five HTLV-IIb4 were found. Interestingly, in the BDs group two HTLV-IIa0 and one HTLV-IIb4 were detected. It should also be noted that 82% of the IVDUs and 50% of the prisoners were coinfected with HIV, while all the BDs were HIV negative. These data indicate that HTLV-IIb4 is the predominant genotype in Italian IVDUs and prisoners, while the significant variability observed in the BD HTLV-II isolates could be due to the different source of infection among this group. PMID- 9208420 TI - Human coronavirus polyadenylated RNA sequences in cerebrospinal fluid from multiple sclerosis patients. AB - Human coronaviruses, represented by the two prototype strains HCV-OC43 and HCV 229E, are important human respiratory pathogens, also associated with necrotizing enterocolitis. Two previous studies, one describing the electron microscopic observation of doughnut-shaped particles, resembling coronaviruses, in a perivascular inflammatory lesion of brain tissue taken at autopsy from a multiple sclerosis patient, and the other one reporting the isolation of coronaviruses from the brains of two multiple sclerosis patients, suggested the possible association between coronaviruses and human demyelinating diseases. We analysed polyadenylated RNAs extracted from cerebrospinal fluid of twenty randomly selected multiple sclerosis patients and ten patients with other neurological diseases (medullary atrophy, Parkinson's disease, polyneuropathy, senile dementia, headache and toxic polyneuropathy) by reverse transcription and polymerase chain reaction searching for HCV-OC43 and HCV-229E sequences. By hybridization analysis of amplification products, we detected HCV-OC43 polyadenylated RNAs in ten specimens of patients with multiple sclerosis. Furthermore, we found positive hybridization signals for HCV-OC43 in the other neurological diseases, except for the toxic polyneuropathy specimen. Positivity for HCV-229E was observed in seven specimens of multiple sclerosis cerebrospinal fluid; one headache cerebrospinal fluid and the medullary atrophy specimen also resulted positive for HCV-229E. Moreover, using a solid phase technique, we report for the first time the sequence of a cDNA fragment derived from RNA extracted from cerebrospinal fluid of multiple sclerosis patient, belonging to the open reading frame which codes for the HCV-OC43 nucleoprotein N. Furthermore, cDNA sequences revealed the presence of a mixed viral population. PMID- 9208421 TI - Detection of JC and BK viral genome in specimens of HIV-1 infected subjects. AB - Human polyomaviruses JC and BK are ubiquitous in healthy human adults, persist as latent viruses and can be reactivated in the immunodeficient host giving different pathologies. Due to the experimental evidence of their potential oncogenicity and neurotropism, as well as to the enhanced viral production induced by co-infection with HIV-1, a possible role of these polyomaviruses has been suggested in AIDS-associated progressive multifocal leucoencephalopathy (PML) and Kaposi's sarcoma. JCV and BKV DNA was detected by PCR in urine and in peripheral blood mononuclear cells (PBMC) using primers specific for structural (VP1) and regulatory (R) regions. In HIV-positive subjects BKV and JCV sequences were found respectively in 8.1% and 31.6% of urine samples whereas in PBMC the positivity increased to 22.8% for JCV and in 51.1% for BKV. Our results indicated that, at DNA level, the presence of BKV and JCV in urine and PBMC was higher in HIV-1 positive subjects than in HIV-1 negative subjects and that, in contrast with JCV, BKV positivity was inversely related to blood CD4-level. Intravenous drug users (IVDU) showed significant increases in both BKV and JCV positivity, while an increased JCV viruria was found in homo-bisexuals compared to heterosexuals. The high prevalence of viral DNA in PBMC of both healthy and HIV positive individuals agrees with the hypothesis that lymphocytes may represent a viral latency site permitting the establishment of virus persistence in affected organs, or a vehicle for the spread of the infection to different tissues. PMID- 9208423 TI - Isolation and characterization of cytopathic strains of rotavirus from hares (Lepus europaeus). AB - Ten cytopathic rotavirus were recovered from stools of leverets affected by an enteric syndrome. The ten isolates, examined by means of ELISA and SDS-PA-GE, were identified as group A rotaviruses. PMID- 9208422 TI - Cytomegalovirus infection in an Italian population: antibody prevalence, virus excretion and maternal transmission. AB - Between 1987 and 1994, the prevalence of antibodies to Cytomegalovirus (CMV) was determined by ELISA in 19,043 subjects in the population of Parma (Northern Italy). The overall prevalence was 71.8%. The age specific prevalence increases starting from 28% in two year-olds to 95.7% in 45-54 year-old subjects. Profiles of antibody production during primary and recurrent infection were analyzed and correlated with virus presence in clinical samples showing correspondence between virus excretion and increasing IgG levels. A longitudinal study of CMV infection was undertaken in 1045 pregnant women and their babies. Rate of infection during pregnancy was 2.34% and rate of congenital infection was 0.57%. Results also indicate that mothers are the major source of perinatal infection (contaminated genital secretions and milk) and confirm the usefulness of monitoring antibody status and virus excretion of mother and neonate at birth. PMID- 9208424 TI - A strain of calicivirus isolated from lions with vesicular lesions on tongue and snout. AB - In December 1992, 17 African lions and 7 Siberian tigers in a Safari park in Japan became sick with characteristic clinical symptoms of acute vesicular formations on tongue and snout. The disease was highly contagious since all of these animals showed similar symptoms within two days after the onset of the first case. Swabs were taken from affected animals in rubbing tongues, snouts and some from rectums. Cytopathic viruses were isolated on CRFK cell culture by virological tests. The physicochemical property of a representative virus strain, named Arthur/L, isolated from a male lion was identified as a member of Caliciviridae. However, seroneutralization test indicated that this virus strain was antigenically distinct from Japanese isolates of feline caliciviruses used for comparison. Viral capsid proteins of the present isolate, Arthur/L, and of a feline calicivirus, strain FC7, were compared in an electrophoresis in SDS-PAGE gel. The major viral capsid polypeptide of them were proved to be significantly different in molecular weight. The polypeptide of FC7 was estimated to be ca. 63 KDa whereas that of Arthur/L consisted of 2 components of ca. 65 and 62 KDa. The viral proteins of these two strains were also proved to be distinct by an immunoblotting test. PMID- 9208425 TI - Viral susceptibility of a newly established cell line derived from the peritoneal cavity of BALB/C mouse. AB - Viral susceptibility of a newly established cell line, named KMP, derived from the peritoneal cavity of BALB/C mouse is described. The cells were originally cloned from the in vitro culture of ascites of the mouse injected with Ehrlich ascitic tumor cells in advance. The electrophoretic pattern of cellular DNAs, extracted from KMP, normal BALB/C mouse spleen, and Ehrlich tumor cells respectively were compared after triple digestions with restriction endonucleases. This cell line was proved to be of mouse origin, but not the sub line of Ehrlich tumor cells. The strains of Coxsackie virus B group, swine enterovirus, influenza virus, encephalomyocarditis virus, vesicular stomatitis virus, and Aujeszky's disease virus were able to multiply well in the cell line with considerably high infectious titers in showing clear CPE and circular plaques. PMID- 9208426 TI - Direct examination of subgingival plaque from a diseased periodontal site using confocal laser scanning microscopy (CLSM). AB - Material taken directly from a periodontal site was investigated using immunofluorescence, acridine orange staining and confocal laser scanning microscopy (CLSM). Porphyromonas gingivalis was tracked by a specific polyclonal antibody and its pronounced occurrence in inflamed as compared to non-inflamed areas was demonstrated. Further accompanying microorganisms were counterstained with acridine orange which could provide information on the viability of individual cells. Optical sections by laser microscopy revealed the spatial arrangement of the investigated material. The combination of specific staining and CLSM allows a detailed microbiological investigation of clinical material obtained directly without cultivation. PMID- 9208427 TI - Taxonomy and identification of periodontopathogenic bacteria and related species. AB - The tumultuous evolution of oral and periodontal microbiology has focussed the attention of many researchers on the necessity to clarify the taxonomic position and identification criteria of putative periodontopathogens, in order to elucidate the role played by each species in the pathogenesis of periodontal disease. Many of the most important periodontopathogens recently underwent a radical reclassification process that may create some confusion and certainly deserves an accurate analysis aimed at focussing the actual situation of these bacteria. The taxonomic evolution and identification criteria of species of the genera Bacteroides, Prevotella, Porphyromonas and Actinobacillus is here analyzed in detail to clarify this recent evolutive taxonomic process, and to explain the importance of molecular studies for both taxonomic and identification purposes in this field. PMID- 9208428 TI - Asynchronous meiotic progression in porcine oocytes matured in vitro: a cause of polyspermic fertilization? AB - The progression of meiosis in porcine oocytes matured in vitro was examined and the effect of maturation interval on the incidence of polyspermic fertilization and embryonic development in vitro was investigated. In Experiment 1, it was found that oocytes selected for in vitro maturation varied considerably in terms of their meiotic progression. Approximately half of the oocytes were already undergoing germinal vesicle breakdown at the start of maturation while the remainder were at the intact germinal vesicle stage. These two populations of oocytes developed to the metaphase-II stage by 24 h and 36 h of maturation respectively. Maturation for 40 h and 44 h did not increase the percentage of oocytes at metaphase-II stage observed at 36 h. In Experiment 2, reducing the maturation interval from 44 h to 36 h did not affect the percentage of oocytes penetrated but did decrease the rate of polyspermic fertilization (5% v. 34%; P < 0.05). In Experiment 3, development of the embryos produced in vitro was assessed after culture for seven days. Reducing the maturation interval from 44 h to 36 h decreased the percentage of embryos developing to the 8-cell (9% v. 18%; P < 0.05), morula (3% v. 10%; P < 0.01), and blastocyst (1% v. 8%; P < 0.001) stages. These results suggest that maturation for 44 h gives rise to a population of 'aged' oocytes which is susceptible to polyspermic fertilization. PMID- 9208429 TI - Are ovine Leydig cells able to aromatize androgens? AB - The conversion of testosterone into oestradiol by ovine Leydig cells cultured in vitro was studied using the non-steroidal aromatase inhibitor CGS 16949A. Additionally, aromatase activity was detected by immunohistochemical staining of cultured Leydig cells or cryosections. The cells were obtained from testes of Polish Mountain rams 5-6 months old (immature) or 12-15 months old (mature). Leydig cells were cultured alone (controls) or incubated for 6 h in the presence of testosterone. Aromatase inhibitor was then added to the cultures which were incubated for a further 18 h. After a 24-h incubation period, testosterone and oestradiol secretion were determined by testing the culture medium using radioimmunological methods. The addition of testosterone to the culture medium enhanced oestradiol synthesis, suggesting that exogenous testosterone could also be aromatized to oestradiol by ovine Leydig cells in vitro. In the presence of CGS 16949A, the conversion of testosterone to oestradiol was significantly suppressed in a dose-dependent manner. All Leydig cells obtained from testes of mature rams and stained immunohistochemically were positive for aromatase, whereas Leydig cells from immature males were negative. The localization of immunoreactive aromatase appeared to be dependent on the age of the donor ram. It is suggested therefore, that mature Leydig cells in the ram are not only the site for testosterone synthesis, they are also capable of converting androgens into oestrogens. PMID- 9208430 TI - Early cleavage to formation of the unilaminar blastocyst in the marsupial Antechinus stuartii: ultrastructure. AB - The development of Antechinus stuartii from the 2-cell stage to the blastocyst stage in vivo was examined by routine transmission electron microscopy. The 2-8 cell stages had a similar organization of organelles, whereas the 16- to 32-cell stages had pluriblast cells and trophoblast cells forming an epithelium closely apposed to the zona pellucida. Specialized cell-zona plugs were formed at the 8 cell stage, and primitive cell junctions appeared in later conceptuses. The cytoplasmic organelles included mitochondria, lysosomes, aggregates of smooth endoplasmic reticulum, lipid and protein yolk bodies and fibrillar arrays, possibly contractile in function. Nuclei had uniformly-dispersed dense chromatin. Nucleoli of 2-4-cell conceptuses were dense, compact and fibrillar, and those of 8-cell conceptuses and later conceptuses were finely granular and became progressively reticulated. The embryonic genome is probably not switched on before the 8-cell stage. Sperm tails were detected in cells in several early conceptuses. The yolk mass had the same organelles as cells. Centrioles were discovered for the first time in marsupial conceptuses. These were prominently situated at a spindle pole in a 32-cell blastomere and were associated with a nucleus and sperm tail at the 4-cell stage. It is very likely that the paternal centrosome is inherited at fertilization and perpetuated in Antechinus embryos during cleavage. PMID- 9208431 TI - Endothelium-derived relaxing factor as a mediator of bradykinin-induced perinatal pulmonary vasodilatation in fetal sheep. AB - Studies in vivo in fetal sheep have shown that bradykinin is released following oxygenation of the lungs and is at least partly responsible for normal pulmonary vasodilatation in the transition from fetal to extrauterine life. Part of this action involves secondary release of prostaglandin I2 (PGI2). In various adult vessels, bradykinin also stimulates the release of a powerful endothelium-derived relaxing factor (EDRF). Studies in vitro were designed (using a modification of the bioassay cascade superfusion technique) to determine whether non-PGI2-related perinatal pulmonary vasodilatation is mediated by an EDRF. Superfused, precontracted, endothelium-denuded strips of fetal sheep thoracic aorta and the maternal sheep main pulmonary artery served as detectors of an EDRF released from isolated, perfused fetal sheep pulmonary arteries. Bradykinin, in the presence of indomethacin to block PGI2 synthesis, caused perfused fetal pulmonary arteries to release an EDRF, which generated a dose-dependent relaxation (24% for 1.0 microM, 16.8% for 0.1 microM, and 10% for 0.01 microM bradykinin). Thus, bradykinin can produce perinatal pulmonary vasodilatation via a mechanism involving the endothelium-dependent synthesis of an EDRF. PMID- 9208432 TI - Comparative studies of conceptus-endometrial interactions in Large White x Landrace and Meishan gilts. AB - Uterine and conceptus function were compared in mated Meishan (MS) gilts and Large White x Landrace (LW x L) gilts (n = 18 breed-1) on Days 11-15, inclusive, after oestrus. Comparisons of individual blastocysts recovered on Day 11 and Day 12 revealed a higher overall embryo survival in MS gilts than in LW x L gilts (100.7 +/- 5.0% v. 69.5 +/- 12.1%; P < 0.05). Embryo survival was higher on Day 11 than on Day 12 (98.6 +/- 6.0% v. 71.6 +/- 12.6%; P < 0.05), with most of the embryo loss between these days occurring in LW x L gilts. MS conceptuses secreted less oestradiol-17 beta and radiolabelled protein on both days, but these differences were not significant. The within-litter variability in the secretion of oestradiol-17 beta and radiolabelled protein by individual conceptuses did not differ significantly between the breeds. Concentrations of epidermal growth factor in uterine rinsings were lower in MS gilts than in LW x L gilts on all days studied. However, two-dimensional polyacrylamide gel electrophoresis and fluorography did not provide evidence of additional breed differences in the profile of endometrial secretory proteins. Consistent temporal changes in the profile of the conceptus secretory proteins were observed among all LW x L gilts and among most of the MS gilts. However, conceptuses cultured from 2 of 5 Day-12 MS gilts secreted a major basic protein which was not evident in other conceptus cultures until Day 14. Similarly, antiviral activity was detected in some cultures of Day-13 MS conceptuses, but was absent from all Day-13 LW x L conceptus cultures. The results also revealed a positive relationship between ovulation rate and the incorporation of radiolabel into endometrial secretory proteins, which was independent of breed. PMID- 9208433 TI - Metabolism of lactate by mature boar spermatozoa. AB - Boar sperm oxidatively metabolized fructose, glucose, glycerol, glycerol 3 phosphate and lactate to CO2 but pyruvate produced only small amounts of CO2 and this was almost completely prevented when endogenous glycolytic metabolism was inhibited. Lactate was the preferred substrate over fructose, glycerol and glycerol 3-phosphate and when lactate was offered in the presence of pyruvate, lactate was preferentially oxidized to CO2. The rate of oxidation of fructose, glycerol and glycerol 3-phosphate was approximately halved in the presence of equi-molar concentrations of lactate and the metabolism of lactate was progressively decreased in the presence of increasing concentrations of mersalyl, an inhibitor of lactate transport. Sperm maintained a high energy charge potential when incubated with lactate as substrate in the presence or absence of bromopyruvate, an inhibitor of endogenous glycolytic metabolism. This evidence confirms that it is lactate, rather than pyruvate, that enters the mitochondria thereby constituting a lactate-pyruvate transport system in these cells for regenerating cytoplasmic nicotinamide adenine dinucleotide (NAD+). Electrophoretic examination of the lactate dehydrogenase isozymes from sperm and several other tissues of the boar showed that sperm contained almost entirely an isozyme which was not present in the other tissues. PMID- 9208434 TI - SRY and karyotypic status of one abnormal and two intersexual marsupials. AB - An intersexual agile wallaby (Macropus agilis) with a penis, a pouch and four teats had a sex-chromosome constitution of XXY in lymphocytes and cultured fibroblasts; the sex-determining region Y (SRY) gene was present, consistent with the presence of a testis. An intersexual eastern grey kangaroo (Macropus giganteus) with a small empty scrotum and no penis, and an abnormal red kangaroo (Macropus rufus) with no penis, pouch or teats, both had XX sex-chromosome complements; the SRY gene was not present, consistent with testis absence. The agile wallaby and grey kangaroo described here provide further evidence that scrotal development in marsupials is independent of the Y chromosome. The cause of the abnormalities in the XX individuals cannot be determined until candidate genes are identified. These animals provide a basis for further genetic studies into marsupial intersexuality and sex differentiation. PMID- 9208435 TI - Development of the lymphoid tissues of the tammar wallaby Macropus eugenii. AB - A study has been made of the development of four lymphoid tissues from birth to maturity in the tammar wallaby Macropus eugenii--the cervical and thoracic thymus, lymph nodes and gut-associated lymphoid tissue (GALT). The development of these tissues in the tammar wallaby is similar to that in two other marsupials, the quokka Setonix brachyurus and the Virginian opossum Didelphis virginiana. Lymphocytes were first detected in the cervical thymus of the tammar at Day 2 post partum and in the thoracic thymus at Day 6. They were subsequently detected in lymph nodes at Day 4 and in the spleen by Day 12 but were not apparent in the GALT until around Day 90 post partum. By Day 21, the cervical thymus had developed distinct areas of cortex and medulla and Hassall's corpuscles were apparent. The maturation of other tissues followed with Hassall's corpuscles in the thoracic thymus by Day 30 and nodules and germinal centres in the lymph nodes by Day 90. Measurement of immunoglobulin G concentrations in the serum of young animals indicated a rise in titre around Day 90 post partum, correlating with the apparent maturation of the lymphoid tissues. PMID- 9208436 TI - Effects of progesterone withdrawal on uterine secretion of prostaglandin F2 alpha in response to oxytocin in ewes. AB - Two experiments were conducted to determine if withdrawal of progesterone during the luteal phase of the oestrous cycle affected the ability of the ovine uterus to secrete prostaglandin F2 alpha (PGF2 alpha) in response to oxytocin. In Experiment 1, 18 ewes were ovariectomized on Day 9 and Day 12 after oestrus. Ewes were subdivided into three treatment groups (n = 6 per group): Group-1 ewes underwent sham surgery; Group-2 ewes received oestradiol (OVX + O); and Group-3 ewes received oestradiol + progesterone (OVX + O,P). Oxytocin was administered to each ewe on Days 10, 13 and 15 after oestrus. Concentrations of 13, 14-dihydro-15 keto-PGF2 alpha (PGFM) were determined in samples of jugular venous blood for 2 h after oxytocin challenge. The magnitude of the PGFM response 24 h after ovariectomy was greater (P < 0.1) in ewes from which progesterone had been withdrawn (OVX + O) than in ewes in which progesterone was maintained (intact controls and OVX + O,P). Therefore, progesterone appears to exert an inhibitory effect on uterine secretory responsiveness to oxytocin which is removed by progesterone withdrawal. In Experiment 2, ewes were ovariectomized on Day 11 and assigned to 1 of 4 treatment groups (n = 6 per group): Group 1, no steroid replacement (OVX); Group 2, oestradiol replacement (OVX + O); Group 3, progesterone replacement (OVX + P); or Group 4, progesterone + oestradiol replacement (OVX + O,P). Ewes received oxytocin on Day 12 and Day 15. On Day 12, uterine secretory responsiveness to oxytocin was greatest in ewes in the OVX + O group (P < 0.1). Responsiveness was low in ewes in the OVX group, as it was in ewes in both groups that received progesterone replacement. Therefore, the increase in uterine secretory responsiveness to oxytocin following progesterone withdrawal is dependent on oestradiol replacement. PMID- 9208437 TI - Efficient selection of preimplantation transgenic embryos by an improved procedure using Dpn I-Bal 31 digestion and the polymerase chain reaction. AB - Efficient selection of preimplantation transgenic embryos by an improved method after pronuclear injection of exogenous DNA is described. The method is based on subjecting DNA extracted from the embryos to restriction enzymes as well as the polymerase chain reaction (PCR). The incorporated procedure included recovery of the digested DNA with glassmilk before PCR, which markedly enhanced the rate of accurate detection of transgenic embryos. When exogenous DNA sequences in the mouse embryos were not integrated into the genome they were digested with both Dpn I and Bal 31, and subsequent PCR analysis generated DNA fragments of the injected DNA sequence in only 1.5% of cases examined. However, DNA extracted from mouse embryos containing the transgene sequences integrated into the genome evaded digestion by both enzymes and yielded transgene-specific PCR products in 68.6% of the embryos tested. When bovine embryos were used, sequences of the endogenous haemoglobin gene used as a control genomic DNA sequence were protected from enzyme digestion (PCR products in 70.5% of the embryos examined); by contrast, the non-integrated injected sequences were almost completely eliminated by the same treatment (PCR products in 1.4% of the embryos examined). It is suggested that this method might be useful for the selection of transgenic embryos before embryo transfer, thereby reducing the number of recipient females required. PMID- 9208438 TI - Lack of correlation between conceptual karyotype and maternal response to placentation. AB - The placental karyotype was correlated with the morphology of the placental bed in miscarriages. Abnormal placentation was as likely to be seen in euploid conceptions as in aneuploid conceptions, whereas normal placentation was seen with both euploid and aneuploid pregnancies. No consistent chromosomal abnormality was found with abnormal placentation. Since abnormal placentation is implicated in the pathogenesis of preeclampsia and miscarriages, these findings may be useful with respect to the localization of a candidate 'preeclampsia gene' by identifying a specific chromosome associated with abnormal placentation. PMID- 9208439 TI - Professor Geoffrey Donald Thorburn AO, FAA. 2 February 1930-28 October 1996. PMID- 9208440 TI - Generating biological models through gene transfer to domestic animals. AB - Transgenic domestic animals remain infrequently used as models for biochemical, biomedical or pharmaceutical studies. The difficulty in obtaining these animals and the cost of their maintenance explains this situation. This review briefly summarizes the different techniques of gene transfer, targeted or not, and also the techniques for constructing vectors for transgene expression. A few examples of domestic animal models are also reported. PMID- 9208441 TI - Pathogenic role of gastric Helicobacter sp in domestic carnivores. AB - As a result of phylogenic studies using new molecular biology techniques and fundamental experimental studies, we now know more about helicobacteria in domestic carnivores, their morphologic characteristics, their taxonomia and more important we know more about their ecological niche. Few clinical studies have been carried out, but the ones that have been undertaken are interesting in that they confirm the extensive prevalence of Helicobacter infections in domestic carnivores and underline their role in the genesis of the inflammatory gastropathies observed in these species. Finally, recent observations have demonstrated the ubiquitous character of these helicobacteria by showing their presence in the stomach of man, dog and cat. This ubiquitous character has led some scientists to consider the potential zoonotic risk of the human infection by Helicobacter heilmannii, felis or pylori. PMID- 9208442 TI - The pig as a model in liver xenotransplantation. AB - Xenotransplantation is considered to be the best solution to the critical shortage of human donors. Despite its phylogenetic distance, the pig appears to be the most appropriate source of organs for transplantation in humans. Its anatomy and physiology are similar to that of man, it can be raised in specific pathogen free environments and it is available in large quantities. The immunological barrier remains to be overcome, however. Considerable progress in the pathogeny of xenogenic rejection has led to the development of therapeutic strategies and the prospect of clinical xenografting appears realisable in the near future. PMID- 9208443 TI - Sensitization of the bovine mammary gland to Escherichia coli endotoxin. AB - The effect of repeated infusions of Escherichia coli endotoxin on the acute phase response in the bovine mammary gland was assessed through the concentrations of tumor necrosis factor alpha (TNF-alpha) in milk. Four clinically normal lactating cows received two intramammary infusions of E coli endotoxin (33 micrograms) 24 h apart in the same mammary quarter. Along with the second infusion, the cows received one dose of endotoxin in the contralateral quarter. Milk was collected at varying intervals before and after infusion and TNF-alpha concentrations were determined by ELISA. Following the first infusion at 0 h, the mean concentrations of TNF-alpha augmented from undetectable concentrations to a maximum of 0.4 ng/mL at 4 h and declined to below 0.04 ng/mL at 24 h, the time of the second infusion. In the quarters challenged twice, the increase in TNF-alpha concentrations was abrupt, culminating at 11.7 ng/mL 6 h later (at 30 h). The increases in TNF-alpha concentrations were similar in the contralateral quarters infused once. TNF-alpha concentrations in the control, uninfused quarters of infused cows remained undetectable (< 0.04 ng/mL). Despite the low TNF-alpha response following the first infusion, mean somatic cell counts increased markedly, being only slightly lower than after the second infusion (10(7)/mL and 5 x 10(7)/mL at 8 h and 32 h, respectively) in the quarters challenged twice. After the first infusion, none of the cows developed fever, but following the second infusion, rectal temperature increased markedly, culminating 6 h after the second infusion. These results show that an infusion in one quarter of an amount of endotoxin sufficient to induce a pronounced cell recruitment but insufficient to induce a marked TNF-alpha secretion following the first infusion sensitized not only that quarter but also the contralateral one to a second infusion with the endotoxin. It is thus possible that sensitization of the whole udder follows a first contact with a moderate dose of endotoxin in one quarter. PMID- 9208444 TI - Use of an experimental chicks model for paratuberculosis enteritis (Johne's disease). AB - Developments in the diagnosis and treatment of paratuberculosis is constrained by the lack of an experimental animal model. To investigate this problem conventional chicks immunodepressed by a Cyclophosphamide injection and concurrent inoculation of Infectious Bursal Disease Virus (IBDV) were infected with Mycobacterium paratuberculosis and kept for 4 months. The immunodepressed chicks eliminated mycobacteria with their faces from the first month until the third month and developed typical intestinal lesions of mycobacterial infection characterized by aggregation of macrophages with monocytes and lymphocytes. Diarrhoea was absent. The number of lymphocytes decreased by about 80%. The serological tests carried out with Complement Fixation test were negative. For the positive bacteriology and typical granulomatous lesions, the conventionally reared chicks proved to be a useful laboratory model for reproduction of Johne's disease. PMID- 9208445 TI - Antigenic variability of porcine reproductive and respiratory syndrome (PRRS) virus isolates. Influence of virus passage in pig. AB - In order to study the antigenic variability of porcine reproductive and respiratory syndrome virus (PRRSV), 18 European and Canadian field isolates were analysed with a panel of 15 monoclonal antibodies (MAbs) raised against five different European and one Canadian PRRSV isolates. The antigenic pattern of these isolates was used to infer their phylogenetic relationships. Isolates which had the same reactivity pattern were gathered together so that five antigenic profiles were analysed. The pairwise distances between these groups were defined based on antigenic pattern differences. Two main antigenic groups were obtained, discriminating between the European and the Canadian populations, as illustrated by the great distance observed between European and Canadian isolates (D = 0.5619 +/- 0.0625) compared to the distance between European isolates (D = 0.1524 +/- 0.0735). The distance matrix allowed also the construction of a tree diagram. Bootstrap analysis was performed to test the confidence in the branching. The tree diagram confirmed the distinction between the European and the Canadian PRRSV populations. Antigenic variability between an isolate and its progeny recovered after one or two passages in vivo was examined on six isolates. It was restricted to the GP3 protein of the virus. PMID- 9208446 TI - Abomasal nematodes in goats from the subtropical island of Grand Canary (Spain). AB - The prevalence of gastric nematodes in 151 goats on Grand Canary Island is around 56%, with a mean burden of 691 worms/animal. No significant differences of prevalence and intensity were found between the four isoclimatic areas of the island [dry-desert (DD); dry-steppe (DS); temperate-mild (TM); temperate-cold (TC)]. Five nematode species were identified, the most commonly-occurring being Teladorsagia circumcincta (observed in 65.8% of parasitized animals) and Trichostrongylus axei (51.9%). The distribution of the two species showed opposing trends: T circumcincta was more prevalent in the coastal areas (DD), diminishing in frequency closer to the centre of the island (TC), while the reverse was true of T axei. The other species identified were Haemonchus contortus, T trifurcata and Camelostrongylus mentulatus; their ranges were restricted to certain areas (H contortus in DS and TM; T trifurcata in DD, DS and TM; and C mentulatus in DD). To analyse the parasite association under natural conditions of an unusual nematode in goats, C mentulatus, with the usual parasites a principal component analysis (PCA) was used to assess the overall behaviour of the nematode community and to examine the Euclidean distances of the parasite associations. The mean Euclidean distances obtained for C mentulatus showed a tendency to a positive association which has also been observed for the other abomasal nematodes. PMID- 9208447 TI - Susceptibility of wild rabbits (Oryctolagus cuniculus) in the United Kingdom to rabbit haemorrhagic disease (RHD). AB - Adult wild rabbits from the southern UK, previously unexposed to rabbit haemorrhagic disease (RHD), were experimentally challenged with a UK strain of the virus in laboratory conditions. Initial serum antibodies were measured by an haemagglutination inhibition test and all seropositive rabbits, with reciprocal titres > 10, were protected against fatal infection. These results are consistent with the behaviour of laboratory and commercially bred rabbits in similar circumstances, and are relevant to consideration of the overall impact of RHD in wild populations. PMID- 9208448 TI - Characterization of acetylcholinesterase secreted by the trichostrongyle nematode parasites of ruminants. AB - Acetylcholinesterase (AChE) secreted by seven different ruminant trichostrongyles was studied in vitro. AChE activity was particularly high (84 and 160 x 10(-3) M.g-1.min-1) in the excretion products of Nematodirus spathiger and N battus, moderate (3 and 5 x 10(-3) M.g-1.min-1) for Trichostrongylus colubriformis and T vitrinus and low (0.9 x 10(-3) M.g-1.min-1) for Teladorsagia circumcincta. No activity was observed with Haemonchus contortus and Cooperia curticei. At 4 degrees C, 80% of AChE activity was maintained over 72 h except for T circumcincta where a loss of 50% was observed after 24 h. At 37 degrees C, N spathiger and T colubriformis maintained an activity over 72 h, but for the other species, a loss of 50% was observed after 24 h. The molecular weights of the AChE from the different species, estimated by gel filtration (Sephadex S300HR), ranged between 64 and 150 kDa. The coefficients of sedimentation estimated by sucrose density gradient ranged between 4.8 S and 7.8 S and corresponded to a monomeric hydrophilic form (G1). For T vitrinus, an amphiphilic form was suspected. PMID- 9208449 TI - Descriptive epidemiology of captive cervid herds in Michigan, USA. AB - A study was designed to determine the species composition, disease period prevalence, and utilization of preventive practices in captive cervid herds in Michigan. This is the first description of cervid farming in the United States. Data for the 12 months preceding the study were collected by means of a mail questionnaire conducted from March 3 through June 28, 1993. Completed questionnaires were returned by 228 of 362 (63%) farms. Study respondents reported ownership of a total of 4972 (80.9%) white-tailed deer (Odocoileus virginianus), 766 (12.5%) elk (Cervus elaphus canadensis), 284 (4.6%) fallow deer (Dama dama), 114 (1.9%) sika deer (Cervus nippon), 6 (0.1%) red deer (Cervus elaphus), 4 (< 0.1%) axis deer (Axis axis), and 2 (< 0.1%) caribou (Rangifer tarandus). The respondents provided disease data for 5493 captive cervids. The most frequent categories of illness in captive cervids (as determined by the period prevalence rates for the 12 months preceding the study) were injuries (1.9%), respiratory disorders (1.1%), foot and leg problems (1.0%), stress due to handling or transport (0.8%), and dystocia (0.8%). The most frequent causes of death were injuries (1.0%), respiratory disorders (0.8%), stress due to handling or transport (0.7%), and unknown causes (0.7%). Use of anthelmintics was reported by 173 of 219 (79%) farms. Only 13% [28/215] vaccinated their cervids. One hundred and fifty-nine of 219 farms reported having a veterinarian who provides cervid consultation. Services provided by these veterinarians include general consultation (58.5% [93/159]), treatment of injuries (27.7% [44/159]), anthelmintic administration (25.2% [40/159]), issuance of health certificates (19.5% [31/159]), diagnosis and treatment of illnesses (17.6% [28/159]), vaccination (13.8% [22/159]), disease diagnosis (treatment provided by farmer) (8.8% [14/159]), foot care (3.8% [6/159]), and other purposes (ie, necropsy, dystocia, antler removal) (11.3% [18/159]). PMID- 9208450 TI - Sequences of terminal non-coding regions from four dengue-2 viruses isolated from patients exhibiting different disease severities. AB - We have determined the 5' and 3' non-coding regions (NCR) of four dengue-2 viruses isolated from dengue patients with different clinical severities in Nakhon Phanom, Northeastern Thailand in 1993. The results were compared to prototype dengue-2 strains and were found to have highest homology with the New Guinea C strain. The sequence of the 5' NCR was completely conserved among all 4 isolates and were identical to the sequence of the New Guinea C strain. Homology of the 3' NCR sequence of the four isolates with the prototype strain ranged from 97.3 to 97.8%. Isolate ThNH-p11/93 from a mild dengue fever case showed the highest divergence from the prototype strain and the rest of the isolates from severe hemorrhagic cases, (1.11%). This includes a change in triad 297-299 nucleotides from the 3' terminus. Computer predicted secondary structures showed that isolate ThNHp-11/93 had significant structural differences from the other three isolates at this region. PMID- 9208451 TI - Human papillomavirus is more prevalent in first trimester spontaneously aborted products of conception compared to elective specimens. AB - In this study the possible role of human papillomaviruses (HPV) in spontaneous abortions is addressed by assaying for HPV DNA in first trimester spontaneous and electively aborted products of conception materials enriched for chorionic villi. The presence of HPVs was measured by polymerase chain reaction (PCR) amplification and DNA dot blot hybridization using an internal probe. The "broad spectrum" HPV primers were directed to amplify E6/E7 junction sequences, while the probe was of an HPV-16 sequence with significant homology to HPV-6/11. The quantity and quality of isolated DNA was also analyzed and compared by observing the PCR amplification of a cellular sequence from the human beta-globin gene. Fifteen of the 25 spontaneous samples (60%) were found to be positive for HPV E6/E7 sequences. In comparison, only 3 of the 15 elective samples (20%) were positive. This is the first study of HPV in fetal materials to incorporate material from elective abortions as a control group. Although confounding contamination from the cervix and vagina can't be ruled out, these data are significant and strongly suggest that HPVs are elevated in spontaneously aborted products of conception. Furthermore, these results suggest the possibility that HPVs may be etiologic agents of at least some spontaneous abortions. PMID- 9208452 TI - Baculovirus expression of proteins of porcine reproductive and respiratory syndrome virus strain Olot/91. Involvement of ORF3 and ORF5 proteins in protection. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) is a new arterivirus that has spread rapidly all around the world in the last few years. The genomic region containing open reading frames (ORFs) 2 to 7 of PRRSV Spanish isolate Olot/91 was cloned and sequenced. The genomic sequence shared 95% identity with Lelystad and Tubingen isolates and between 61-64% with the ORF7 region of the American isolates. ORFs 2 to 7 were inserted into recombinant baculoviruses downstream of the polyhedrin promoter. Only ORFs 2, 3 5 and 7 were expressed in insect cells as detected by PRRS-specific pig antisera. To analyze the immunogenicity of these proteins and their ability to confer protection, Sf9 cells infected with recombinant baculoviruses expressing ORFs 3, 5 and 7 gene products were used to immunize pregnant sows, either individually or in combination. The results obtained indicate that ORFs 3 and 5 gene products could be major candidates for the development of a vaccine against PRRS since they conferred 68.4 and 50% protection, respectively, as evaluated by the number of piglets born alive and healthy at the time of weaning. In addition, piglets born to sows immunized with ORFs 3 and 5 proteins were seronegative to PRRSV after weaning, indicating absence of viral replication. ORF7 is the most immunogenic protein of PRRSV, but the antibodies induced in sows are non-protective and may even interfere with protection. PMID- 9208453 TI - A major antigenic domain of hantaviruses is located on the aminoproximal site of the viral nucleocapsid protein. AB - Hantavirus nucleocapsid protein has recently been shown to be an immunodominant antigen in hemorrhagic with renal syndrome (HFRS) inducing an early and long lasting immune response. Recombinant proteins representing various regions of the nucleocapsid proteins as well as segments of the G1 and the G2 glycoproteins of hantavirus strains CG18-20 (Puumala serotype) and Hantaan 76-118 have been expressed in E. coli. The antigenicity of these proteins was tested in enzyme immunoassays and immunoblots. These studies revealed that human IgG immune response is primarily directed against epitopes located within the amino acid residues 1 to 119 of the amino terminus of viral nucleocapsid proteins. This fragment was recognized by all HFRS patient sera tested (n = 128). The corresponding enzyme immunoassays proved to be more sensitive than the indirect immunofluorescence assays. Furthermore, the majority of bank vole monoclonal antibodies raised against Puumala virus reacted specifically with this site. A recombinant G1 protein (aa 59 to 401) derived from the CG 18-20 strain was recognized by 19 out of 20 sera from HFRS patients. PMID- 9208454 TI - Sequence and phylogenetic analysis of the cytoplasmic inclusion protein gene of zucchini yellow mosaic potyvirus: its role in classification of the Potyviridae. AB - The cytoplasmic inclusion (CI) gene of a Singapore isolate of zucchini yellow mosaic virus (ZYMV-S) was sequenced and compared with CI of 14 other potyviruses. In addition to the consensus sequence GAVGSGKST of nucleotide binding motif (NTBM) which is implicated as a membrane-binding component of the RNA helicase complex, five other conserved motifs were found. Phylogenetic trees were constructed from sequence data for the CI and the coat protein. Similar branching patterns obtained from both CI and coat protein analyses suggests that phylogenetic relationship among potyviruses can be determined using the CI. We propose that phylogenetic analysis of CI gene may be used as an alternative approach for the study of evolution within the family Potyviridae. PMID- 9208455 TI - Analysis of the fusion protein gene of the porcine rubulavirus LPMV: comparative analysis of paramyxovirus F proteins. AB - Complementary DNA clones representing the fusion (F) protein gene of the porcine rubulavirus LPMV were isolated and sequenced. The F gene was found to be 1,845 nucleotides long containing one long open reading frame capable of encoding a protein of 541 amino acids. The cleavage motif for F0 into F1 and F2 is His-Arg Lys-Lys-Arg. A sequence comparison and a phylogenetic analysis was performed in order to identify possible functional domains of paramyxovirus fusion proteins and also to classify the porcine rubulavirus. The F gene of LPMV is most closely related to the human mumps virus and simian virus type 5 F genes, and is therefore classified into the rubulavirus genus. A coding region for a small hydrophobic protein was however not found between the F and hemagglutinin neuraminidase (HN) genes as previously found in both SV5 and mumps. PMID- 9208456 TI - Baculovirus expression of the respiratory syncytial virus fusion protein using Trichoplusia ni insect cells. AB - Respiratory syncytial virus (RSV) is a major viral pathogen responsible for severe respiratory tract infections in infants, young children, and the elderly. The RSV fusion (F) protein is highly conserved among RSV subgroups A and B and is the major protective immunogen. A genetically-engineered version of the RSV F protein was produced in insect cells using the baculovirus expression system. To express a secreted form of this protein, the transmembrane domain was eliminated by removing the region of the gene encoding 48 amino acids at the C-terminus. Production of the truncated RSV F protein (RSV-Fs) was compared in two different insect cell lines, Spodoptera frugiperda (Sf9) and Trichoplusia ni (High Five). The yield of RSV-Fs secreted from High Five insect cells was over 7-fold higher than that from Sf9 insect cells. Processing of the RSV-Fs protein was also different in the two insect cell lines. N-terminal sequencing demonstrated that while most of the RSV-Fs protein secreted by High Five cells was correctly processed at the F2-F1 proteolytic cleavage site, most of the RSV-Fs protein secreted by Sf9 cells was unprocessed or incorrectly processed. Antigenicity of the major RSV F neutralization epitopes was maintained in the RSV-Fs protein secreted from High Five cells. The RSV-specific neutralizing antibody titres in the sera of cotton rats immunized with the RSV-Fs protein were equivalent to those in the sera of animals intranasally inoculated with live RSV. Animals immunized with either live RSV or the immunoaffinity purified RSV-Fs protein from High Five cells were completely protected against live virus challenge. PMID- 9208459 TI - 6-L-alanineferrirubin, a ferrichrome-type siderophore from the fungus Aspergillus ochraceous. AB - The molecular structure of 6-L-alanineferrirubin tetradecahydrate, [Fe(C41H64N9O16)].14H2O, has been determined in order to confirm its chemical structure. The structural results show that the presence of an alanine in place of a serine or a glycine at position 6 in the cyclic hexapeptide has very little effect on the conformation of the 18-membered ring or on the geometry of the octahedral iron coordination. PMID- 9208460 TI - 1,8-bis(hydroxymethyl)anthracene. AB - Anthracene-1,8-dimethanol, C16H14O2, crystallized in the centrosymmetric space group P21/n. The anthracene core is nearly planar and shows good agreement with the anthracene core of anthracene-1,8-dicarboxylic acid. The geometric disposition of the hydroxymethyl groups and the values of the hydrogen-bond parameters are strikingly similar to those in the naphthalene analog. As in that structure, each hydroxyl group participates both as a donor and acceptor in an infinite zigzag chain of hydrogen bonds which propagates along the [010] direction. Moreover, the disposition of the entire molecule in the cell of the present compound is also strikingly similar to that of the naphthalene analog in its cell. PMID- 9208457 TI - A random DNA sequencing, computer-based approach for the generation of a gene map of molluscum contagiosum virus. AB - The genome of Molluscum contagiosum virus (MCV) has a high G + C content, which largely differs from those of vaccinia virus (VAC) and other characterized poxviruses. This has precluded the use of DNA hybridization for the identification of MCV genes and the further establishment of the virus genetic map. To circumvent this problem, we have partially sequenced clones containing virus restriction endonuclease fragments, which were derived by either single or double-digestion of genomic DNA from the subtype I of MCV. The DNA sequences were translated and used to search protein data bases. This analysis resulted in the finding of high-scoring matches to data base entries, including forty-five VAC genes. In addition, MCV-specific sequences that encoded protein domains of known function (i.e. DNA J domain) were found. The locations of MCV clones were inferred from the presumed colinearity of both MCV and VAC genomes, and further confirmed by PCR technology. The data presented here led to the construction of a partial genetic map of MCVI, which revealed that the order and orientation of a large number of MCV genes were equivalent to those of their VAC homologues. The conserved gene arrangement was apparently disrupted in the terminal regions, where MCV sequences showing homologies with the VAC counterparts were not found. PMID- 9208458 TI - Identification and DNA sequence analysis of the Marek's disease virus serotype 2 genes homologous to the thymidine kinase and UL24 genes of herpes simplex virus type 1. AB - The thymidine kinase (TK) gene has been used as a safe and convenient locus for expression of heterologous proteins in some alphaherpesviruses including herpesvirus of turkeys (HVT) antigenically related to Marek's disease virus (MDV) serotypes 1 (MDV1) and 2 (MDV2). In MDV2 strain HPRS 24 genome, genes equivalent to the TK and UL24 homologues of herpes simplex virus type 1 were identified and sequenced. The MDV2 UL24 gene overlaps the 5' end of the TK gene in a head-to head orientation. The predicted region encoding for the MDV2 TK gene is 1,056 nucleotides, corresponding to a polypeptide of 352 amino acids in length. Putative nucleotide- and thymidine-binding sites were identified within the predicted amino acid sequence. The predicted region encoding for the UL24 gene is 948 nucleotides, corresponding to a polypeptide of 316 amino acids in length. By northern blot analyses using MDV2 TK- and UL24-specific DNA probes, four transcripts of approximately 7.8, 5.0, 3.5, and 1.1 kb for the TK gene, and a transcript of 3.8 kb for the UL24 gene were detected in MDV2-infected cells. Alignment of the amino acid sequence of MDV2 TK homologue with those published for TK homologues of other MDV serotypes showed 73.9% (MDV1 vs. MDV2), 58.2% (MDV1 vs. HVT), and 56.8% (MDV2 vs. HVT) identities. Comparison to other alphaherpesvirus TK homologues revealed amino acid sequence homologies varying from 34.5% to 27.8%. The putative MDV2 UL24 homologous protein had identity with the well conserved five motifs among alphaherpesviruses. PMID- 9208461 TI - 1-(1,3-Benzothiazol-2-yl)-2-(2-bromo-5-nitrophenyl)ethanone. AB - The title compound, C15H9BrN2O3S, was isolated as an unexpected product from the reaction of the anion of sodium 2-(1,3-benzothiazolyl)ethanonitrile with alpha,2 dibromo-5-nitrotoluene. Its structure features a benzothiazole fragment and a bromo- and nitro-substituted phenyl ring linked by a methyl ketone group. The dihedral angle between the benzothiazole and phenyl rings is 103.7 (2) degrees. The benzothiazole fragment is planar, with a maximum deviation of 0.021 (2) A. The nitro group is slightly rotated out of the phenyl-ring plane, with a O(2) N(2)-C(14)-C(15) torsion angle of 16.4(7) degrees. PMID- 9208463 TI - Batch lot variability in permeation through nitrile gloves. AB - Many factors should be considered in the selection of chemical protective clothing, but the majority of selections are based on manufacturers' permeation data composed of average results for three replicates; usually no information about variability is provided. It was hypothesized that variability across batch lots might be considerable, and that variability may be due to cure factors that may vary from one site to another within the same company. Glass transition temperature (Tg) has been demonstrated to be an indicator of cure, and so its relationship to permeation parameters was examined. Steady state permeation rate, breakthrough detection time (BDT), cumulative permeation at 125 minutes (ASTM F1407), and Tg (ASTM E1356) were measured for two makes of nitrile gloves presumably in four batches. Tg was not related to any of the permeation parameters even though batch-to-batch variability was statistically significant for all parameters except BDT. A comparison with recent ASTM round-robin results indicates that some of the variability may be due to the method; however, manufacturer quality control must be suspected as the major source of variability based on the results of this study. PMID- 9208462 TI - A clerodane diterpene with antibacterial activity from Ajuga lupulina. AB - The structure of a new diterpene, C30H46O11, with antibacterial activity against Pseudomonas aeruginose and Escherichia coli, isolated from the fresh whole plants of Ajuga lupulina (Labiatae) was established to be 2 beta-hydroxy-2 methylbutanoyl-3 alpha-lupulin (3-deoxy-14,15-dihydro-2-hydroxy-15 methoxycaryoptinol 2-methylbutanoate), by means of X-ray crystallographic analysis. The present study confirms that the two six-membered rings are in ideal chair conformations. PMID- 9208464 TI - Physical load as risk factor for musculoskeletal complaints among tank terminal workers. AB - In a cross-sectional study, the relationship between physical load and musculoskeletal complaints was examined among operators (n = 77), office workers (n = 52), and miscellaneous workers (n = 15) in a tank terminal company. Information about history of musculoskeletal complaints, individual characteristics, and working conditions in past and present was obtained by a standardized interview. Assessment of physical load was performed by direct observation of awkward postures and forceful exertions during normal activities. Workers filled out a diary regarding the executed tasks during one shift. Physical load was also modeled by linking the average physical load of workers performing a particular task with the task distribution of each worker. The modeling approach was beneficial to estimate infrequent characteristics of physical load, whereas the observations were better for assessing frequent exposure to awkward trunk postures. After adjustment for age and other confounders the observed work time with trunk flexion and rotation was associated with the occurrence of back pain. The modeled work time with arms raised above shoulder showed a relationship with elbow pain, and the modeled work time with forceful exertions over 100 N showed relationships with elbow pain and wrist pain. The described relationships demonstrate that the assessment strategy chosen in a particular study strongly influences random measurement error of physical load, and that inappropriate strategies may mask true associations between physical load and musculoskeletal complaints. Modeling strategies offer the opportunity to assess physical load in dynamic work environments. PMID- 9208465 TI - Participatory ergonomics in a red meat packing plant. Part II: Case studies. AB - Three ergonomics-related case studies are presented to demonstrate the problem solving method used by two participatory ergonomics teams. The problem-solving method was adapted from principles related to quality management (e.g., participation, structure, a scientific approach, and decision by consensus). The first two steps of the problem-solving method were related to identification and evaluation of the problem; the latter three steps were related to solution development, implementation, and evaluation. The problem evaluation process included the collection of background, exposure, and effects data. Solution development following evaluation of the problem started with a brainstorming session, then discussion to select interventions by consensus. The format for presenting the case studies was intended to be concise and visual with the intent of effectively documenting the teams' problem-solving processes. PMID- 9208466 TI - A test chamber for experimental hydrogen fluoride exposure in humans. AB - An inhalation chamber was built to perform experimental studies with hydrogen fluoride (HF), other gases, and particulate matter. The present study sought to describe a new gas delivery system and the distribution and concentration of HF gas in the chamber. The aluminum chamber has a volume of 19.2 m3 and a variable ventilation rate of about 1 to 10 air changes per hour. The negative pressure difference between the chamber and outside air can be regulated from 0 to 300 Pa. HF was fed at concentrations of up to 4000 mg/m3 directly into the ventilation duct feeding the chamber through openings with diameters as small as 50 microns, oriented opposite to the airflow. Gas flow was varied from about 0.1 dm3/min at a pressure of 4 atm. The dilution factor of HF concentration from cylinder to chamber was on the order of 10(3) to 10(4). The standard deviation (SD) of the HF concentrations at a fixed measurement point during a 1-hour test was typically 0.05 mg/m3 at a time-weighted average (TWA) concentration of 2.66 mg/m3. The SD of the TWA HF concentrations at six locations in the chamber was typically 0.05 mg/m3 and 0.29 mg/m3 at 0.61 and 3.46 mg/m3, respectively. Human exposure could be predicted from calculations based on ventilation data, gas flow, and observed ratio between calculated and measured concentrations. When the target exposure concentration was 1.5 mg/m3, the measured mean exposure concentration was typically 1.54 mg/m3 (range: 1.4-1.7 mg/m3, SD 0.09 mg/m3, n = 8). The chamber is well-suited for inhalation studies in humans. Chamber atmosphere was controlled and has proved to be stable and homogeneous, even in tests with HF, a highly reactive gas in the class of superacids. PMID- 9208467 TI - Maximum acceptable forces for repetitive ulnar deviation of the wrist. AB - The purpose of this experiment was to quantify maximum acceptable forces for ulnar deviation motions of the wrist at various repetition rates. Subjects grasped a handle with a power grip and moved it through a 1.40 rad (80 degrees) ulnar deviation wrist motion (similar to a knife cutting task). A psychophysical methodology was used in which the subject adjusted the resistance on the handle and the experiment manipulated or controlled all other variables. Two series of experiments were conducted. Thirteen subjects completed the first series, which investigated repetition rates of 15 and 20 motions per minute. Eleven subjects completed the second series, which investigated 15, 20, and 25 motions per minute. Subjects performed for 7 hours per day, 5 days per week, for 4 weeks in the first series and 5 weeks in the second series. The subjects were instructed to work as if they were on an incentive basis, getting paid for the amount of work they performed. Symptoms were recorded by the subjects during the last 5 minutes of each hour. The results are presented and compared with maximum acceptable forces for wrist flexion and extension. PMID- 9208468 TI - Sex differences in aggression: does social representation mediate form of aggression? AB - In contrast to the usual pattern of higher male aggression, girls have been shown to exceed boys on frequency measures of indirect aggression. Women also tend to view aggression in terms of an expressive social representation whereby it is seen to result from loss of self-control, in contrast to men who tend to describe it as an instrumental act aimed at exerting control over others. Sixteen items measuring different forms of aggressive behaviour were given to 105 undergraduates together with Expagg, a psychometric measure of social representation of aggression. Factor analysis of the aggression items revealed three scales: direct (verbal and physical), indirect instrumental and indirect expressive aggression. The only aggression scale showing a significant sex difference was indirect expressive aggression on which women scored higher than men. There was also a significant sex difference on Expagg with women showing a more expressive representation of aggression. However the point biserial correlation between sex and indirect expressive aggression was not diminished when expressive representation of aggression was partialled out. It is argued that indirect expressive aggression (involving bitching and avoiding) fails to show a relationship with social representation because it lacks the formal requirements of intentional harm and consequently is not an act of 'aggression'. PMID- 9208469 TI - The limitations of trust in intimate relationships: constructions of trust and sexual risk taking. AB - This paper challenges the assumption that trust is a generally desirable, static, measurable attitude of the individual. A reconceptualization of trust as situationally specific, negotiated and purposeful social action is proposed. Data presented in this paper are based on a series of semi-structured interviews with 16 heterosexual individuals. The interview agenda included questions about respondents' sexual behaviour within the context of HIV/AIDS. All interviews were tape-recorded and transcribed. Parker's (1992) version of the discourse analytic method guided the analysis of the transcripts. Three discursive constructions of trust were identified: trust-as-security, trust-as-symbolic-practice and trust-as social-regulation. All of these were employed by respondents in order to justify sexual risk taking. The paper traces the ways in which these constructions are deployed and discusses their implications for relationship dynamics. Further research into the ways of 'doing trust', both publicly and privately, is called for. PMID- 9208470 TI - Swine manure cleanup criteria calculation for odor causing volatile organic compounds based on manure-to-ventilation air exposure pathway. AB - A mathematical model was derived to calculate swine manure cleanup criteria for odor causing volatile organic compounds (VOCs) so that acceptable total dose would not exceeded through the inhalation of ventilation air in swine housing for workers and pigs. A hypothetical scenario was used which assumed that subsurface contaminant in swine manure diffuses through a layer of manure-air interface then is swept into the ventilated airspace via advection, where long-term inhalation of contaminant was assumed to occur. The philosophy of the transport model is to incorporate the age distribution of contaminated air and a first-order decay of contaminant sources into the diffusion model for simulation of air concentrations of VOCs and total exposure dose. A closed-form solution is presented to allow a series of numerical experiments for investigating the effects of adsorption characteristics between manure gas and manure, the mean age of contaminated air, effective diffusivity, and degradation coefficient on total dose. Swine manure cleanup criteria based on non-exceedence of the total hazardous dose corresponding to an acceptable risk from indoor inhalation of four selected VOCs of p-cresol, hexane, toluene, and xylene were calculated in a typical pig unit. The model can be used in the future to compute the relative effectiveness of VOCs filtration systems and/or altered ventilation rates on the VOC exposure problem in animal housing. PMID- 9208471 TI - The complexation of Cu(II) by anthropogenic fulvic acids extracted from composted urban and livestock wastes. AB - The fulvic acid (fua) fractions of two samples of composted solid wastes [urban (urfua) and livestock (lsfua) wastes], commercialized to be used in agriculture as organic correctives or fertilizers, were analyzed for their affinity towards Cu(II) at pH = 6. Molecular fluorescence spectroscopy (synchronous mode) was used to monitor the quenching caused by the complexation upon addition of Cu(II) to fua. Spectral data were preprocessed by a chemometric self-modeling mixture analysis method (SIMPLISMA) to detect the number of different types of fluorescent binding sites that exist in each fua, their spectra and the corresponding quenching profiles [fluorescence intensity as function of the total Cu(II) concentration]. From the analysis of the quenching profiles, the amount of binding sites (CL) and the corresponding conditional stability constants (K') were calculated. Both fua samples have approximately CL = 0.21 mmol/g and the logarithms of K' are 4.21(3) and 4.51(8), respectively for urfua and Isfua. The differences detected between these fua samples and those extracted from natural soils can be attributed mainly to the relatively small humification extent of the present anthropogenic fua samples. PMID- 9208472 TI - The subchronic toxicity of acridine in the rat. AB - The subchronic toxicity of acridine was investigated in rats following dietary exposure at 0, 1, 10, 100 and 500 ppm for 13 weeks. The growth rate and food consumption were not affected by treatment and no clinical signs of toxicity were observed. There was a slight but significant decrease in spleen weight, both in absolute terms and as a percent of body weight, in the 500 ppm males and a slight increase in absolute thymus weight in the females of the same dose group. Both hepatic ethoxyresorufin O-deethylase (EROD) and pentoxyresorufin O-dealkylase (PROD) activities were slightly, but significantly, elevated in females in the 500 ppm dose group. No haematological or other biochemical changes were observed. Females also displayed dose-related increases in inorganic phosphate and uric acid levels. Treatment-related histopathological changes were seen in the thyroid, liver and kidney and included hepatic anisokaryosis and vesiculation of nuclei and glomerular adhesions, reticulin sclerosis and nuclear pyknosis in the kidney. Residue data showed a dose-dependent accumulation of acridine in liver, kidney and adipose with the highest concentration being found in the fat of the 500 ppm dose group. Based on these data, the no observable adverse effect level of acridine was judged to be 100 ppm or 12 mg/kg bw/day. PMID- 9208474 TI - The asteroid body of lobomycosis. AB - Lobomycosis is a dermal mycosis produced by Loboa loboi. It gives rise to dermal granulomas in which giant cells phagocytose an overwhelming number of fungi. Intracytoplasmatic asteroid bodies (AB) have been observed in some giant cells. Their nature is unknown and they have been confused with the sporotrichotic AB. We studied 84 skin biopsies from 53 patients with lobomycosis, 7 by electron microscopy (EM). Immunohistochemistry with a polyclonal anti-Sporothrix schenckii antibody was performed in five biopsies. We found AB in the giant cells in 22 of the 84 biopsies on HE staining. They appeared as single eosinophilic intracytoplasmic structures surrounded by a clear empty space. This clear space did not appear when the biopsy was fixed with OSO4, thus indicating that it is lipid in nature, and that the vacuole is an artefact produced by lipid extraction. Under the EM, the AB consist of bundles of dense, filamentous material, dense bodies and myelin structures resembling the image of AB in sarcoidosis. Some giant cells containing AB either phagocytosed only few fungi or did not contain any of them. The AB and the fungi did not react with the anti sporotrichotic antibody, which stained the extracellular AB of sporotrichosis. These two asteroid structures are, therefore, different and unrelated phenomena. PMID- 9208473 TI - Sexual forms of yeasts in clinical samples. AB - The sexual or teleomorphic state of yeasts has only been described in a few clinically involved species, mainly those of the Saccharomycetaceae family. With the aim of gathering information on their incidence in human pathology, a study has been made of a total of 2,135 strains isolated from clinical samples and cultivated in McClary agar. From these, 8 strains in teleomorphic state were identified: Kluyveromyces marxianus [1], Pichia anomala [2], Pichia farinosa [1], Pichia membranaefaciens [1] and Saccharomyces cerevisiae [3]. The two strains of P. anomala were responsible for fungemia; K, marxianus and the two strains of S. cerevisiae produced vaginitis; the other strains were oral cavity colonizers. PMID- 9208476 TI - Mating patterns of dermatophytes of diverse origin in India. AB - The mating patterns of Trichophyton mentagrophytes var. interdigitale (74 isolates) and the Microsporum gypseum complex (17 isolates) of diverse origin and T. rubrum (25 isolates) and T. tonsurans (10 isolates) of clinical origin were studied. The results of the study showed that the teleomorph of the Indian isolates of T. mentagrophytes belong to Arthroderma vanbreuseghemii, undetermined teleomorphs of T. mentagrophytes var. interdigitale (+) mating types, and undetermined teleomorphs of T. mentagrophytes var. interdigitale indeterminate mating types. All the isolates of T. rubrum and T. tonsurans were found to be of the (-) mating type. PMID- 9208477 TI - Fungal biodiversity in the air of Turin. AB - The qualitative fungal composition of Turin's atmospheric environment was surveyed, carrying out a twelve-month study and collecting with a single stage volumetric sieve sampler on Dermasel agar supplemented with 0.4 g l-1 cycloheximide and 0.05 g l-1 chloramphenicol. We isolated 165 species and 2 varieties of mesophilic fungi from 58 genera and 26 thermotolerant species from 12 genera. Penicillium, Aspergillus, Acremonium, Chrysosporium, Scopulariopsis, Malbranchea, Paecilomyces, Phialophora and Cladosporium were in sequence the genera most rich in mesophilic species; Aspergillus, Penicillium, Chrysosporium and Scopulariopsis the most rich in thermotolerant species. Many of the species isolated are rarely or never recorded in the atmospheric environment. Cycloheximide can thus be said to select among airborne fungi, giving a characteristic picture. PMID- 9208475 TI - Purification and characterization of fatty acid synthetase from Cryptococcus neoformans. AB - Fatty acid synthetase has been purified from Cryptococcus neoformans 450 fold to a specific activity of 3.6 units per mg protein with an overall yield of 23%. The purified enzyme contained two non-identical subunits, Mr approximately 2.1 x 10(5) and 1.8 x 10(5). Under optimum conditions, 100 mM KCl and pH 7.5, apparent K(m) values for the substrates were: Acetyl CoA, 19 microM; Malonyl CoA, 5 microM; and NADPH, 6 microM. Product inhibition patterns were determined to be: CoA, competitive versus acetyl CoA and malonyl CoA, uncompetitive versus NADPH; NADP, competitive versus NADPH, uncompetitive versus acetyl CoA and malonyl CoA; Palmitoyl CoA, competitive versus malonyl CoA, noncompetitive versus acetyl CoA and NADPH; Bicarbonate, uncompetitive versus malonyl CoA. These product inhibition patterns are consistent with the multisite ping-pong mechanism previously proposed for the avian fatty acid synthetase complex. The cryptococcal fatty acid synthetase was inhibited by the polyanionic polymers, heparin and dextran sulfate, an effect never before demonstrated for a fatty acid synthetase. This inhibition exhibited a marked dependence on the length of the polymer chain, with dextran sulfate fractions with Mr of 6 x 10(5) and above having Ki values below 100 nanomolar. A model is presented that involves initial binding of the anionic polymer to the enzyme complex at a region of high positive charge density, followed by interaction of the end of the tethered polymer with the catalytic site. This study represents the first purification of fatty acid synthetase from a basidiomycete. PMID- 9208478 TI - Fungi associated with post-harvest rot of black plum (Vitex doniana) in Nigeria. AB - The fungi associated with rot of Vitex doniana fruits (blackplum) were isolated and identified. Aspergillus niger, Botryodiplodia theobromae, Candida spp. Penicillium chrysogenum, Rhizopus stolonifer, Fusarium pallidoroseum F. oxysporum and Mucor mucedo were the primary rot causing fungi in contrast to Cladosporium herbarum and Mucor circinelloides which were just present as secondary colonizers. The rot fungi penetrated mainly through wounds and bruises on the surface of fruits. Mature green fruits were less susceptible to infection than half ripe and fully ripened red fruits. Optimum rot by pathogenic isolates occurred at 25-30 degrees C and relative humidity 72.5-100%. The results of investigation of influence of storage temperatures and relative humidity on the quality of uninoculated healthy fruits are presented and discussed. PMID- 9208479 TI - Fungi associated with herbal drug plants during storage. AB - Samples of sundried herbal drug plant parts stored for sale were purchased from four herbal markets located in Ibadan, Nigeria. The plant parts were analysed for mycoflora associated with their storage. Fungal colony counts ranged from 0.60 x 10(2) to 3.50 x 10(2) within the period of the survey. Twenty eight fungal species were isolated with the species of Aspergillus niger, A. flavus, Fusarium moniliforme, Trichoderma viride, Penicillium expansum and Mucor fragilis being the dominant ones. There were marked differences between the flora obtained on fresh plant parts and those stored for sale. The results obtained showed that herbal drug plant pieces stored for sale in the markets are hazardous for human health. There is therefore the need for some form of quality control and decontamination before they are used for herbal drug preparations. PMID- 9208481 TI - Neuropeptidergic innervation of human nasal mucosa in various pathological conditions. AB - This study aims to investigate the roles of neuropeptides in the pathophysiology of human nasal diseases. By using immunohistochemistry and radioimmunoassay, we detected the presence, distribution and concentrations of the following neuropeptides in human nasal tissue: vasoactive intestinal peptides (VIP), neuropeptide Y (NPY), substance P (SP), and calcitonin gene-related peptides (CGRP). This was performed in human nasal inferior turbinate mucosa from 20 patients with allergic rhinitis, twenty-five patients with chronic hypertrophic rhinitis and 10 patients without any nasal disease conditions. The presence and distribution of NPY. CGRP and SP fibers among the three subject groups displayed no evident differences. VIP fibers were densely stained around the vessels in the allergic group. In contrast, these fibers were more prominently distributed around the submucosal glands of the chronic hypertrophic rhinitis group. The concentration of VIP and SP in human nasal inferior turbinate showed a significant increase in allergic subjects. Thus, VIP may be revelant to the hypertrophic changes of the nasal mucosa. Both SP and VIP may play significant neuroimmunological roles in the pathogenesis of allergic rhinitis. PMID- 9208480 TI - Characterization of polymorphic mononuclear cell in porcine Actinobacillus pleuropneumonia. AB - The polymorphic mononuclear cells, arranged in whorling or palisading pattern, were usually found in the lung of Actinobacillus pleuropneumoniae-infected pigs. In order to understand the origin and characteristics of these cells, specific pathogen free pigs were intratracheally inoculated with Actinobacillus pleuropneumoniae at a concentration of 5 x 10(6) CFU, then, sacrificed at 6, 12, 24 and 48 hours later. The cells in the alveolar spaces were observed with light and electron microscope, and cytochemically analyzed for acid phosphatase, alkaline phosphatase, alpha-naphthyl acetate esterase, and Sudan black B stains respectively. The results revealed that a lot of neutrophils were observed in the alveolar spaces at the early stage after inoculation. Twenty four hours later, polymorphic mononuclear cells abundantly appeared. Enzyme cytochemical findings indicated that some of the polymorphic mononuclear cells were macrophages, in which, acid phosphatase and alpha-naphthyl acetate esterase were detected, and others were type II pneumocytes which were positively stained with alkaline phosphatase and Sudan black B. Ultrastructural observation found that many lysosomes appeared in the macrophages' cytoplasm, and type II pneumocyte contained many lamellar bodies. Conclusively, it could be suggested that the polymorphic mononuclear cells were derived from macrophages and type II pneumocytes by cytochemical and electron microscopic examinations. PMID- 9208482 TI - Comparative study of the effects of capsaicin on the contractility of normal and spinal cord injured human bladders. AB - In order to investigate the effects of capsaicin on human detrusor contractility in both normal and spinal cord injury (SCI) bladders, a detrusor contractility study was performed in 10 normal and 8 SCI patients using isolated muscle strips. Eight bladder muscle strips were harvested from each patient undergoing surgery. Four strips were treated with capsaicin of 1-1000 microM, and electrical field stimulation and bethanechol were applied to the strips before and after capsaicin administration. The other four strips were pretreated with 40 nmole [D-Arg1, D Trp7,9,Leu11]-Substance P (spantide) and then were underwent the same procedure. The results showed that capsaicin induced a dose-dependent increase in muscle tension on the human detrusor in both normal and SCI bladders. After treatment with varying concentrations of capsaicin for 10 minutes, low doses of capsaicin partially depressed detrusor contractility under both electrical and bethanechol stimulation while high doses of up to 1000 microM almost totally blocked detrusor contractility. The initial contractile effect of capsaicin was higher in normal bladders but the final depressant effect did not show any difference between normal and SCI bladders. With addition of spantide, the initial contractile effect and the final depressant effect of capsaicin remained the same, indicating that the contractile effects of capsaicin were not mainly through NK receptors but directly on muscle cells. Consecutive application of capsaicin to the same strip could not reproduce the contractile response. After washing free of capsaicin, the detrusor contractility under electrical stimulation and bethanechol was not reversible. A direct neurotoxic or cytotoxic effect could be found after high concentration capsaicin administration. In treating patients suffering from detrusor hyperreflexia using intravesical capsaicin instillation, this effect should be considered to prevent irreversible damage to the urinary bladder. PMID- 9208483 TI - Weight bearing influence on knee joint bony contact movements: an in vivo video fluoroscopy study. AB - In order to understand the effects of body weight-bearing on knee joint bony contact movements, a video-fluoroscopic digitizing system with in vivo human knee extension-flexion motions of 12 healthy adults under body weight-bearing and non weight-bearing conditions was studied. These 12 subjects were equally separated into two groups consisting of a body weight-bearing group and a non-weight bearing group. Video-fluoroscopic images were digitized to get three parameters from the knee joint bony geometry. These three parameters were the radius of rotation, the are length of rotation, and the contact point of the tibiofemoral joint, and they were used to decide the knee joint bony contact status of the sliding, spinning and rocking motions. The results showed that the knee bony contact movements under body weight-bearing conditions had about 4 times the incidence rate of the sliding motion under non-weight-bearing conditions. The incidence rate of the sliding motion was greatest when the knee flexion was less than 30 degrees. The knee bony contact movements under non-weight-bearing conditions had a larger spinning motion incidence rate and smaller rocking motion incidence rate than they did under weight-bearing conditions. The larger spinning motion incidence rate when the knee joint flexion was greater than 60 degrees. In conclusion, the body weight-bearing factor should be considered in studying knee joint bony contact movements. PMID- 9208484 TI - The relationship between spermatozoa and epithelium of the female genital tract during sperm storage in the greater yellow bats (Scotophilus heathi): the light and electronmicroscopic observations. AB - The present paper describes the relationship between spermatozoa and the epithelium of the female genital tract during sperm storage in greater yellow bats, Scotophilus heathi. All the female bats collected from mid-January till ovulation in early March showed the presence of spermatozoa with their heads orientated towards the epithelial lining of the female genital tract. During this period, the epithelial cells of the uterine horn showed extensive endoplasmic reticulum (ER), numerous mitochondria and well developed Golgi complexes. In females inseminated during December, the spermatozoa were not arranged linearly and showed signs of degeneration. During December, the uterine epithelial cells contained only a few small mitochondria and less developed rough ER. The leucocytes and Langerhans cells were frequently seen in the epithelial lining during the course of sperm storage especially at the site of phagocytised sperm heads. PMID- 9208485 TI - Investigation of refolding condition for Pseudomonas fluorescens lipase by response surface methodology. AB - Response Surface Methodology (RSM) was used to investigate optimal refolding conditions of Pseudomonas fluorescens lipase which was expressed as inclusion body in E. coli. Three interacting factors, protein concentration, pH and guanidine hydrochloride (GdnHCl) concentration were selected as the variables of RSM. The protein concentration did not affect the refolding yield within the selected range (50-340 micrograms ml-1) at low temperatures of 4 and -15 degrees C, but it was a critical factor at 25 degrees C and refolding yield significantly decreased with increasing protein concentration. The pH and GdnHCl were significant factors in all experimental conditions. But there was no trends of optimal pH depending on temperature and additives, just showing the optimum at neutral pH. Glycerol shifted optimal GdnHCl concentration to higher side, while arginine to lower side. Therefore, it was concluded that glycerol and arginine deprives and supplements GdnHCl, respectively. In this study, RSM made it possible to investigate successfully the optimal conditions of in vitro refolding and to elucidate interactions between refolding factors with a minimum number of experiments. Under the optimal condition determined by RSM, approximately 90% refolding yield was obtained and it was a 30% increase over the conventional method. PMID- 9208486 TI - Metabolic flux distributions in recombinant Saccharomyces cerevisiae during foreign protein production. AB - A stoichiometric flux balancing analysis was applied to the recombinant yeast cultivation to examine the cellular physiology and relationship between the production of heterologous protein and metabolic fluxes. The fluxes in the metabolic pathway within a recombinant S. cerevisiae grown on galactose alone or mixtures of galactose and ethanol medium were calculated. It is found that an amplification of the PP (Pentose Phosphate) pathway activity resulted in an improvement of the foreign protein expression and cell yield on ATP. The carbon source used for biosynthesis from TCA cycle in the exponential growth phase was 2 and 5-fold higher, respectively, as compared with that in the late exponential growth phase and stationary phase in batch culture with galactose minimum medium. The metabolism of ethanol together with galactose in the recombinant cell looks like increasing the flux from Acetyl-CoA to TCA cycle, and amplifying the flux directing the synthesis of various kinds of precursors such as amino acids and nucleic acid which are necessary for production of a foreign protein. Metabolic flux distribution analysis also shows that the ATP synthesis rate under substrate level phosphorylation in the mixed carbon source cultivation was lower than that in the sole carbon source (galactose) during the expression of foreign protein. However, the total ATP production rate was higher in the mixed carbon source cultivation. PMID- 9208487 TI - Chemotherapy of ovarian carcinoma. PMID- 9208488 TI - Management of ovarian endodermal sinus tumor. AB - OBJECTIVE: To evaluate the role of combination chemotherapy and the optimal cycles of treatment in improving the prognosis of ovarian endodermal sinus tumor, and to study the relationship between the type of surgical management and the outcome of the disease. PATIENTS AND METHOD: Sixty-three patients with ovarian endodermal sinus tumor were divided into 3 groups according to the postoperative chemotherapy they had received. Group 1 (37 patients) patients were treated with at least 6 cycles of VAC (vincristine, actinomycin-D and cytoxan) or 4 cycles of PVB (cysplatin, vincristine and bleomycin). Group 2 (17 patients) patients were treated with VAC in less than 6 cycles or PVB in less than 4 cycles. Group 3 (9 patients) patients received no VAC or PVB but some other drugs like TSPA, 5FU, MTX and cytoxan in various combinations. The sustained remission rates and survivals were compared among these 3 groups. For patients with full courses of treatment with VAC or PVB (Group 1), different types of surgical management were studied about their relationship with the outcome of the disease. RESULTS: The persistent remission rates are 81.8%, 23.5% and 11.8% for group 1, group 2 and group 3 patients respectively (P < 0.001). The survival curve of group 1 is very much different from that of group 2 and group 3 patients. With full courses of chemotherapy with VAC or PVB, it appears that the different types of surgical managements (unilateral vs bilateral adenectomy; with vs without systemic lymphadenectomy; residual tumor < 2 cm vs > 2 cm) did not show definite relationship with the outcome of the disease. CONCLUSION: Combination chemotherapy with VAC or PVB dramatically improved the prognosis of ovarian endodermal sinus tumor but it should be emphasized that the favorable results could be obtained only when the treatment is given on time and in optimal cycles. Although ovarian endodermal sinus tumor is chemosensitive, appropriate surgical treatment is still important, however, the surgical techniques need some further studies. PMID- 9208489 TI - Effect of carboplatin based combination chemotherapy on ovarian cancer. AB - OBJECTIVE: To investigate the effect of carboplatin based combination chemotherapeutic regimen given intraperitoneally and intravenously on ovarian epithelial cancer. MATERIALS AND METHODS: Carboplatin based combination chemotherapy was given intraperitoneally and intravenously to 48 patients with epithelial ovarian cancer. Of them, 10 cases with ascites had received no treatment before, and 38 patients had undergone cytoreductive surgery prior to the study. RESULTS: The results of this study showed that carboplatin based combination chemotherapy could not only reduce the tumor mass but also decrease the amount of ascites (P < 0.05) in patients with ascites. The response rate was 80% (clinical complete response rate was 10%) in untreated patients with ascites. The 3-year survival rate of patients with ovarian epithelial cancer was 92.3% in stage I, and 30% in stage III patients after the cytoreductive surgery. Tumor recurrence occurred during the course of chemotherapy in 15% of stage I patients and 53.3% of stage III patients. CONCLUSION: Carboplatin based combination chemotherapy given intraperitoneally and intravenously has potent effect on untreated ovarian cancer with ascites. But carboplatin based chemotherapy could not overcome the problem of drug resistance. To improve the prognosis of ovarian cancer, much more researches on the mechanisms of drug resistance need to be carried out. PMID- 9208490 TI - Experimental study on the mechanism of cisplatin resistance and its reversion in human ovarian cancer. AB - OBJECTIVE: To explore the mechanism of cisplatin resistance and its reversion in human ovarian cancer. MATERIALS AND METHODS: A cisplatin resistant cell subline of SKOV3, SKOV3/cp, was established, and a xenograft mice model of human ovarian cancer was established by microencapsulation technology. Various biochemical changes and the effects of modulators on the resistance in the model were observed. RESULTS: The intracellular platinum accumulation. Pt-DNA adducts and interstrand cross links of DNA (ISC) in SKOV3 was 5.1, 2.4 and 4.8 times respectively of those in SKOV3/cp cell line. Amphotericin B (AmB) and Novobiocin (NVB) could raise platinum accumulation and Pt-DNA adducts concentration in SKOV3/cp and this resulted in reversion of cisplatin resistance. CONCLUSIONS: The primary factors resulting in SKOV3/cp resistance to cisplatin are the reduction of intracellular drugs and the augmentation of the ability to remove Pt-DNA adducts. AmB and NVB can reverse cisplatin resistance in SKOV3/cp cells. PMID- 9208492 TI - Detection of cytomegalovirus DNA in paraffin-embedded lung tissue specimens using in situ polymerase chain reaction. AB - OBJECTIVE: To develop a possible alternative laboratory diagnostic method, an in situ polymerase chain reaction (ISPCR) for definitive diagnosis of human cytomegalovirus (HCMV) pneumonia in infants. METHODS: The material involved in this study was formalin-fixed paraffin-embedded lung tissue specimens from 6 infants died of histopathologically diagnosed cytomegalic inclusion body pneumonia (CMIBP) and from 4 infants died of other diseases. When DNA extracts from the specimens of these 10 infants were tested by using a previously applied conventional PCR, the specimens from the 6 infants with CMIBP were positive for HCMV DNA, while those from the other 4 infants were negative. In the ISPCR, biotin-11-dUTP was used for labeling the amplified products and streptavidin, biotin-alkaline phosphatase for in situ color development. RESULTS: The results of the study showed specific staining for HCMV DNA in the lung tissue slices from the 6 cases with CMIBP, while no staining was found in the specimens from the 4 infants who died of other diseases. The control ISPCR tests on the HCMV DNA positive specimens without adding either biotin-11-dUTP or Taq DNA polymerase or the primers for HCMV DNA showed negative results. The specific staining for HCMV DNA was seen in bronchial and alveolar epithelial cells, small vascular endothelial cells and leukocytes in the lumens of small blood vessels. CONCLUSIONS: The results suggested that the ISPCR was able to demonstrate both presence of HCMV DNA in the lung tissues and active HCMV infection; therefore the ISPCR may be applicable in definitive diagnosis of HCMV pneumonia when proper specimens such as lung biopsy material or bronchoalveolar lavage is available for the test. PMID- 9208491 TI - Exploration for drug therapy in endometrial carcinoma. AB - OBJECTIVE: To explore the therapeutic effect of three drugs, i.e., hydroxyprogesterone caproate (HPC), Tamoxifen (TMX) and Aminoglutethimide (AG), in the treatment of endometrial carcinoma. METHODS: The patients were collected by double-blind method and classified into 6 groups (3 groups used single drug and 3 groups used combined drugs) at random. The patient was given assigned drug therapy for 10 days according to her group after being admitted to the hospital. Operations were performed 7-10 days after drug therapy. Plasma FSH, LH, E2 and P were measured by standard radioimmunoassay of WHO. ER and PR in cancer tissues were examined by enzyme linked histochemistry. Cancer tissues obtained before and after drug therapy were examined pathologically. RESULTS: The values of FSH, LH and E2 decreased and that of P increased after single drug therapy in the group treated with HPC; and the values of FSH, LH, E2 and P all decreased in the groups treated with TMX and AG respectively (P < 0.05). The values of FSH, LH and E2 decreased to different extent in the groups treated with combined drugs. The changes of FSH and E2 are remarkable (P < 0.05). The value of P increased a little with no statistical difference (P > 0.05). PR and ER usually increased after treatment with TMX, but decreased with HPC or AG, or combined drugs. The pathological changes indicate that the tumors responded to all of the three drugs. The response to TMX was most significant, following which was HPC. CONCLUSIONS: HPC and TMX are two drugs proved to be useful in endometrial carcinoma. AG also can be used to treat endometrial carcinoma. It adds a new drug to the drug therapy for endometrial carcinoma. PMID- 9208493 TI - Abnormal expressions of hepatocellular proteins and extracellular matrix in CCL4 induced liver injury in rats. AB - OBJECTIVE: To evaluate the effects of carbon tetrachloride (CCl4) on the expressions of both intracellular proteins and extracellular matrix and the protective role of taurine. METHODS: Forty Sprague-Dawley rats were divided into five groups: 1.5% taurine, CCl4, CCl4 + 0.5% taurine, CCl4 + 1.5% taurine and control. CCl4 was given ig at a dose of 500 mg/kg body weight, twice a week for 5 weeks. The taurine solution or tap water was given to the rats for drinking. The sections (4 microns) of rat liver were stained with haematoxylin and eosin, immunohistochemistry or colloid silver methods. RESULTS: The animals treated with 1.5% taurine and control had normal liver structures. Light microscopic examination of sections of the liver taken 5 weeks after dosing of CCl4 showed hydropic degeneration of hepatocytes around the central veins and lipid vacuolation in mid-zonal and some periportal hepatocytes. Necrotic cells and neutrophil infiltration around most central veins as well as hepatocytes showing various stages of mitosis indicative of regeneration were observed. These histopathological changes in the animals treated with CCl4 alone showed more extensive and severe lesions than those seen in the animals treated with CCl4 plus taurines. The incidences of PCNA and p53 antigen positive hepatocytes, and the numbers of nuclear apoptotic bodies and AgNORs granules were markedly higher in the CCl4-treated animals than in those protected with taurine. Declined and unevenly distributed expressions of LN and FN were seen in the CCl4 treated rats. CONCLUSIONS: Both the damage of hepatocellular DNA and the abnormal expressions of hepatic extracellular matrix were observed in the CCl4-treated rats. Taurine, in this study, not only inhibited hepatocellular degeneration, necrosis and DNA damage, but also inhibited the lesion of extracellular matrix induced by CCl4. PMID- 9208494 TI - Promotion of differentiation and proliferation of peripheral blood CD34+ cells in vitro by G-CSF. AB - OBJECTIVE: To observe the effect of granulocyte-colony stimulating factor (G-CSF) on differentiation and proliferation of CD34+ cells from peripheral blood in presence of recombinant hematopoietic growth factor (HGF). METHODS: Peripheral blood mononuclear cells mobilized by G-CSF were obtained from patients suffering from carcinoma, preparing for autologous bone marrow transplantation. CD34+ cells were isolated by derivatized polystyrene tissue culture flask which had covalently immobilized soybean agglutinin lectin and was coated with anti-CD34 antibody; and this kind of cells were incubated in liquid culture medium for up to 28 days under the stimulation of combination of growth factors, i.e., stem cell factor (SCF) and interleukin-3 (IL-3) with or without G-CSF. The changes of nucleated cells, colony forming unit-granulocyte and monocyte (CFU-GM), burst forming unit-erythrocyte (BFU-E), colony forming unit-megakaryocyte (CFU-MK), and myeloid-associated markers were evaluated. RESULTS: An increase of nucleated cells (mean 640-fold increase) occurred during culture CFU-GM production is parallel to the nucleated cell production until the 11th day (mean 82-fold increase) in combination of 3 HGF, i.e., G-CSF, IL-3 and SCF. A large number of cells expressing late myeloid markers appeared on the 11th day in suspension culture of CD34+ cells. CONCLUSION: G-CSF was found to synergize with IL-3 and SCF in inducing rapid proliferation of purified CD34+ cells and differentiation to multiple myeloid lineages. The stroma-free, cytokine-driven culture system could achieve a degree of amplification of colony forming cells, suggesting the feasibility of culture of hematopoietic progenitor cells in vitro as an adjunct to hematopoietic stem cell transplantation. PMID- 9208495 TI - Abnormal coronary flow reserve in patients with angina pectoris and hypertensive left ventricular hypertrophy. AB - OBJECTIVE: To assess coronary flow reserve using a computer-assisted method with intracoronary papaverine in patients with angina pectoris. PATIENTS AND METHODS: Coronary arterial diameter, cross-sectional area and blood flow velocity were measured during coronary arteriography before and after intracoronary papaverine in 26 control subjects, 45 patients with significant coronary artery disease (> 50% luminal narrowing), 16 patients with syndrome X and 14 patients with hypertension and left ventricular hypertrophy (LVH). RESULTS: After intracoronary administration of papaverine, proximal diameter, cross-sectional area, blood velocity, flow volume and reserve capacity of both left anterior descending and right coronary arteries were lower in patients with significant coronary stenosis than in the controls. Despite similar changes in diameter and cross-sectional area, the blood velocity, flow volume and reserve capacity of the two vessels were also lower in patients with syndrome X than in the controls. In 14 patients with hypertension and LVH, although the blood velocity and flow volume were augmented for the two arteries and did not differ from those in the controls, the flow reserve was reduced because of higher baseline blood velocity and flow volume. CONCLUSIONS: Coronary flow reserve is reduced in patients with coronary artery disease, syndrome X or hypertensive LVH, which may be related to abnormal changes at different levels of the coronary vasculature or resting flow states. PMID- 9208496 TI - Exercise echocardiography: feasibility and value for detection of coronary artery disease. AB - OBJECTIVE: To evaluate the feasibility and accuracy of exercise echocardiography (Ex-Echo) for the diagnosis of coronary artery disease (CAD) based on coronary angiography (CA). PATIENTS AND METHODS: Forty-seven patients were found to have CAD and examined by upright exercise electrocardiography (Ex-ECG) on a treadmill within two weeks of CA. Before and immediately after exercise, the patients lay on a bed beside the treadmill in the left lateral position and parasternal long and short axis and apical two and four chamber views of the heart were acquired. Pre- and post-echocardiograms were analysed in a side-by-side multiple screen format on the imaging view and hypodynamic wall motion of the left ventricle after exercise was defined as Ex-Echo positive. The sensitivity, specificity and predictive accuracy of Ex-Echo and Ex-ECG were calculated on the basis of CA data. RESULTS: Satisfactory echocardiograms were recorded in 46 patients after exercise. The success rate was 97.8%. Compared with Ex-ECG, Ex-Echo was more sensitive (87.5% vs 62.5%, P < 0.05), specific (92.8% vs 60.0%, P < 0.05) and accurate (89.4% vs 61.7%, P < 0.01). The concord of determining the number of diseased vessels between coronary angiography and Ex-Echo was 90.9% in single vessel disease and 45.0% in multiple vessel disease. Wall motion scoring index, however, was higher in multiple than in single vessel CAD. CONCLUSIONS: Ex-Echo test is feasible and accurate in detecting CAD and wall motion scoring index is probably useful in distinguishing multiple from single vessel CAD. PMID- 9208497 TI - Acute bacterial meningitis in children in Hefei, China 1990-1992. AB - OBJECTIVE: To obtain etiologic and epidemiologic information about bacterial meningitis, especially the H influenza type B (Hib), from a medium-sized city, Hefei, China. METHODS: Data were collected prospectively over 3 years, from 1990 to 1992 by a well-organized group including 13 hospitals. All children with a clinical diagnosis of acute bacterial meningitis were enrolled and the specimens were taken for the etiologic studies. CSF and blood were tested by standard bacteriologic technique. CSF, blood and concentrated urine were tested directly for detection of antigen by countercurrent immuno-electrophoresis (CIE). Data were analyzed by epidemiologic methods. RESULTS: Bacterial culture and CSF Gram's staining were positive only in 13.3% and 11.7%, respectively. Bacterial antigen detection was positive in up to 90% by CIE which was more sensitive than bacterial culture (chi 2 = 67.7, P < 0.005). The annual incidence of acute bacterial meningitis in the city is calculated as 9.3 cases/100,000 children from 1 month to 15 years of age and 19.2 cases/100,000 children from 1 month to 5 years of age. Hib meningitis accounted for 51.7%, N. meningitis (Nm) for 38.3%, and S. pneumoniae (Sp) for 8.3%. There was no significant seasonal variation. Of the patients, 76.7% were children under 5 years of age, and 51.7% under 1 year of age. The case fatality rate was 11.7% for all bacterial meningitis, 9.7% for Hib, 17.4% for Nm and 20% for Sp. A total of 22.6% of survivors suffered from neurological or psychological problems. CONCLUSIONS: Using antigen detection combined with bacterial culture, we could make an etiologic diagnosis in up to 90% of the patients in this group. Hib, Nm and Sp were the predominant pathogens, which was similar to the findings in other countries. Hib was the most common cause of bacterial meningitis, but the incidence was much lower than in most parts of the world. PMID- 9208498 TI - Spike waves and synaptic ultrastructure in human epileptic brains. AB - OBJECTIVE: To find morphological changes of synapses related to the spike discharge by observation on ultrastructures of the EEG spike foci of cerebral cortexes. MATERIAL AND METHODS: The cerebral cortexes were obtained surgically from the EEG spike (recorded by flaky silver electrodes on the surface of cortexes) foci and nonspike areas within the limits of the brain tissue that would be removed in 20 cases of human epilepsy, and were processed for transmission electron microscopic (TEM) observation. RESULTS: In the spike foci, presynaptic terminals of axospinal asymmetric synapses were obviously swollen but postsynaptic spines were relatively unaffected. In these synapses, the synaptic vesicles remarkably decreased in number and accumulated predominantly at the presynaptic membranes or aggregated. Similar changes were observed in the presynaptic terminals of the axospinal asymmetric synapses in the nonspike areas but were of milder degree than in the spike foci. CONCLUSION: Axospinal asymmetric synapses may release a large quantity of excitatory neurotransmitters, which may result in epileptic abnormal electric activities during seizure. PMID- 9208500 TI - A scanning electron microscopic study of trabeculae in osteoporotic femoral head. AB - OBJECTIVE: To investigate osteoclastic resorption in trabeculae of osteoporotic femoral head. METHODS: Osteoporotic femoral heads were collected from 7 aged women with an average age of 72.4 years, who underwent endoprosthetic replacement for intracapsular hip fracture. Femoral head trabeculae from 3 young adults killed in traffic accidents served as control. The two types of specimens were processed and studied under scanning electron microscope. RESULTS: The trabeculae of femoral head formed round or roundish arch structure. The columnar trabeculae of femoral head in the aged women showed overt osteoclastic resorption, manifested in thinning, tapering and perforation, resulting in formation of icicle-like trabeculae, which then became rounded, lost height and eventually turned into small tubercle. As a result, the inter-trabecular space enlarged markedly. Under high magnification, the trabeculae could be discerned oval, narrow oval or spindle-shaped. Howship resorption lacunae, which varied in size, depth and content, but all revealed punched-out margin. During bone resorption, the inorganic and organic components were successively resorbed. In the Howship lacunae and surrounding areas, newly formed collagen fibrils and bone tissues emerged, signifying reversal and new bone formation phases after bone resorption phase. CONCLUSION: Osteoclastic resorption markedly compromises the structural integrity and strength of the trabeculae of the arch structure of the femoral head in the aged women. PMID- 9208499 TI - Surgical treatment of coronary artery fistulae associated with other cardiovascular anomalies. AB - OBJECTIVE: To evaluate retrospectively our surgical experience, techniques and long-term results in 11 patients with coronary artery fistulae associated with other cardiovascular anomalies. METHODS: From January 1980 to April 1995, 11 patients with coronary artery fistulae associated with other cardiovascular anomalies were found among 20,000 open-heart procedures and treated surgically. Besides closure of the fistulae, coronary artery bypass grafting was performed in 5 patients with atherosclerotic coronary artery disease, and aneurysm angioplasty was done in 4 patients with coronary artery aneurysm. Ventricular septal defect and patent ductus arteriosus were closed in one patient and mitral valve replacement was performed in another patient. RESULTS: There were no surgical deaths, but one late death due to acute myocardial infarction occurred 6 months after surgery. The mean follow-up time was 75.7 months and all patients' functional status improved by an average 1.4 judged by the New York Heart Association functional classification. CONCLUSIONS: Coronary artery fistula associated with other cardiovascular anomalies will aggravate symptoms and cause deterioration of heart function. Therefore, evaluation and surgical intervention should be done as soon as possible to restore coronary blood flow and correct concomitant cardiovascular anomalies. The surgical results are excellent with a low operative risk and good long-term results. PMID- 9208501 TI - Treatment of cutaneous T cell lymphoma with low doses of interferon alpha-2b. AB - OBJECTIVE: To evaluate the optimal dose and route of interferon-alpha-2b (INF alpha-2b) given to the Chinese patients with cutaneous T-cell lymphomas (CTCL). PATIENTS AND METHODS: Sixteen Chinese patients with CTCL treated with INF-alpha 2b were evaluated. They were divided into groups according to the TNM Classification: I B (8 patients), II A (2), II B (5) and IV B (1). During induction INF-alpha-2b was given intramuscularly every other day or two times a week, beginning with a dose of 1-3 x 10(6) IU. Depending on tolerance, the dosage could be increased weekly to a maximum tolerated dose of 18 x 10(6) IU (mean, 6 x 10(6) IU/week) with administration two or three times a week. Patients with clinical response were given maintenance dosage from the start. Thereafter the dosage was gradually decreased after disappearance of skin lesions. Three patients were treated by combining intramuscular and intralesional injection of INF. RESULTS: Six patients (I B, 2 patients: II A, 2, II B, 2) achieved complete response (CR). Seven patients (I B, 5 patients) showed partial response. The overall response rate in this group was 81.25%. The prominent initial clinical response in three patients was manifested early at one week after intramuscular treatment with INF and then the skin lesions were resistant to the therapy but disappeared rapidly by combining intralesional injection of INF. The conditions of those patients obtaining CR within 3 to 6 weeks were stable for 1-36 weeks. CONCLUSIONS: This study had demonstrated that the overall response rate in treatment of patients with CTCL was 81.25% and higher (85.7%-87.5%) in the subgroups at early stages I and II of the disease. The dosage (mean dose of 6 x 10(6) IU/week) adopted by us is optimal for the Chinese patients with CTCL. Intralesional injection of INF could be recommended. PMID- 9208502 TI - Recent progress in researches on precancerous lesions of gastric cancer in China. PMID- 9208503 TI - Possibility of observing the changes of cerebrospinal fluid pulse waves as a substitute for volume pressure test. PMID- 9208504 TI - Primary central nervous system lymphoma. A changeable tumor. PMID- 9208505 TI - [Evaluation on the effect of supplementary immunization for poliomyelitis elimination in Hainan province]. AB - We appraised the effect on poliomyelitis elimination programme in Hainan Province, based on data as incidences of poliomyelitis in last years, the routine immunization and the activities on National Immunization Day (NID) and Provincial Immunization Day (PID). The result indicated that the effect of NID in 1993/1994 was remarkable. In Hainan Province, there has been no poliomyelitis case caused by wild strain of poliovirus since 1994. Laboratory data also demonstrated that NID could break the circulation of wild strain of type I poliovirus. We found that routine immunization alone could not effectively eliminate poliomyelitis but other activities of NID must be carried out in order to form an immunity-barrier within a short period. PMID- 9208506 TI - [Supervision and evaluation of vaccination coverage rates in Jiangsu province]. AB - Comprehensively, this paper evaluated the actual vaccination coverage rates in Jiangsu province, using geometric averages to weigh vaccination coverage rates through a) regular and irregular investigation b) conventional report rates and c) registration rates of children at the right age of vaccination. Results showed that the comprehensive method of using vaccination coverage rates for evaluation was better than the conventioned index which had been used before. PMID- 9208507 TI - [Criterion on the screening for acute flaccid paralysis (AFP) surveillance]. AB - Using AFP as the criterion for surveillance of children aged < 15 years on polio eradication programme, the sensitivity is 100%. But specificity is low, with high false positivity. Through analysing all 818 cases of AFP occurred in 1991 in Anhui province, we were trying to find out a more specific criterion for the purpose of screening polio cases from AFP. Among 818 cases of AFP aged < 15 years, most polio and non-polio AFP cases occurred under 5 years. At the onset of paralysis, 87.63% of the polio cases presented with fever, compared with 45.99% of non-polio AFP cases did. There fore we chose age and fever at the onset of paralysis as the criterion for screening tests. In parallel tests, the sensitivity was very high, but the specificity was still low. In serial tests, the combination of age under < 6 and fever at onset of paralysis resulted in a sensitivity of 81.44% and specificity of 65.54%. This results suggested that by screening the cases of AFP aged < 6 years who had fever at the onset of paralysis, the number of sample size under testing can be largely reduced, with a minimal compromise in the sensitivity. As for surveillance purpose, the criterion can be used to identify more critical cases followed by urgent attention. PMID- 9208508 TI - [Serological effect after immunization with rabies vaccine of different doses]. AB - Ninety-one cases bitten by pets as dogs, cats were randomly divided into two groups, and immunized with different doses of rabies vaccine. The antibody levels were detected by IFAT after immunization. Result showed that the immune response of the group who received boosting doses was better than the 5-doses group. 51 cases were examined by IFAT and ELISA. The positive rates of IFAT and ELISA were 72.5% and 64.9%, respectively (P < 0.05). PMID- 9208509 TI - [An epidemiologic study on the aetiological factors of primary liver cancer in Shunde City of Guangdong province]. AB - In order to investigate the aetiological factors of Primary Liver Cancer (PLC) in Shunde City of Guangdong province, 96 clinically diagnosed PLC patients and 144 matched hospital controls were interviewed and their blood samples were examined for HBV, HCV and other seven indices. Monofactorial and multifactorial analyses were fitted by using Non-conditional Logistic Regression model. The findings confirmed the strong association between HBV infection and PLC. Histories of hepatitis and histories of eating raw fish, shrimp or salt fish were also noticed. The history of alcohol intake might have associated with PLC. The present study did not find associations between PLC and drinking water, histories of blood transfusion and injection- or exposure to insecticides. Antibody of HBV surface antigen seemed to be a protective factor with a relative risk of 0.3064 (0.1647-0.5701). The results showed that the combined effects of HBsAg infection and HCV infection were worthy for further study. PMID- 9208510 TI - [A three-year follow-up study of hepatitis C virus infected individuals from Beixie village, Gu-an county, Hebel province, China]. AB - A total number of 42 anti-HCV positive individuals from Beixie village, Gu-an county, was followed for three years to study the dynamic change of alanine aminotransminase (ALT), anti-HCV titers as well as HCV RNA in serum. Research results showed that 18.1% to 26.2% of the individuals exhibited a rise in ALT values while the positive rate of HCV RNA was 80%. The yearly negative seroconversion rate of anti-HCV was only 0.95%. The geometric mean titer (GMT) of anti-HCV was around 160 and showed no significant difference between the paired serum samples collected in 1991 and in 1994. There were no statistical associations between the GMT value of anti-HCV and ALT level as well as HCV RNA in serum. PMID- 9208511 TI - [A genetic epidemiologic study on HBsAg chronic carriers]. AB - 375 Individuals from 23 HBsAg positive families were investigated in the study. The results showed that HBsAg carrier rate among blood relatives was significantly higher than non-blood relatives (P < 0.01); HBsAg carrier rate decreased with the order of the first, second and third degree relatives (P < 0.01); and the rate in the individuals living together with the probands was higher than those living apart (P < 0.01). However, the other two markers of HBV infectivity, anti-HBs and anti-HBc, did not show any differences mentioned above. The results analysed by means of dichotomy and Logistic regression model, showed that blood relationship played an important role in HBsAg carrier state. In addition, the history of sharing living facilities was related to HBsAg carrier state. The average heritability in the first, second and third degree relatives was 79.68%. The analysis of genetic model showed that HBsAg carrier state was corresponded to the characteristic of multifactorial genetic disease, excluding the possibility of genetic disease due to single gene. PMID- 9208512 TI - [A serologic and epidemiologic survey on 150 cases with acute viro-hepatitis]. AB - Though the urban district of Jiaxing City has been one of the high incidence areas of viro-hepatitis in Zhejiang province, there is no systematic research study either on its serology or its epidemiology. A total number of 150 cases from Jiaxing urban district, admitted in Jan. Dec., 1993, was reported in this paper. Among them: HA 45.33%, HB 51.33%, HC 5.33%, HD 2% and HE 8.67%, including single infection, mixed infections and super-infection of different individuals. Sex, age, profession, seasonal distribution and other epidemiologic characteristics were discussed for the parpose of scientific preventive measures and preventive stratagy development. PMID- 9208513 TI - [A serologic study of legionellas in the patients with corpulmonle infection]. AB - By MAT, the serum antibodies to Lp1-Lp14 were measured in fifty patients with infection of corpulmonale admitted in three hospitals in Datong. The antibody positive rate was 38% and the infection rate was 18%. The result indicated that some of the patients with corpulmonle infection were caused by Lp. and the seriousness of Corpulmonle was related to the infection of Lp. An alternative method for the therapy of the infection of corpulmonle was offered. PMID- 9208514 TI - [Impact on the development of extrinsic asthma caused by dust mites exposure, and its sensitization in Jiangsu]. AB - To obtain information on the relationship between dust mites exposure, dust mite sensitization, and the development of extrinsic asthma in Jiangsu province between 1992-1993, a case-control study was carried out. Unconditional logistic regression method was conducted. The results showed that the history of allergy in parents, dust mites exposure, dust mite sensitization, and birth in months with high mite densities were factors which played important roles in the development of extrinsic asthma. The population attributable rate of dust mites exposure was 70.29%. Based on the results, some practical preventive measures were put forward. PMID- 9208515 TI - [Result on the screening of scoliosis among school students in Beijing area]. AB - An investigation of the morbidity of scoliosis has been carried out in Beijing area including both urban and rural area. The survey involved 21759 elementary school students aged 8 to 14. The result showed that the morbidity of scoliosis was 1.06 per cent. Most patients had idiopathic scoliosis while congenital scoliosis possessed only 5.19% of the whole patients. Prevalence in female students was slightly higher than in males. In terms of morbidity of scoliosis, there was no significant difference seen in the areas of study. The use of Adam forward-bending test for original examination in the investigation seemed to be simple and effective. The spine measure ruler is qiute simple in production and easy to handle. Using the ruler, one will get less exposure to X-ray. The film which shows the whloe spinal AP and lateral view would lay a final and ture basis for diagnosis. PMID- 9208516 TI - [An analysis to the focus of (American Journal of Epidemiology) research with bibliometrics methods]. AB - Using bibliometric method, the author counted the citation of papers published in American Journal of Epidemiology in the last 3 years. The highly cited papers and books were presented and the focus of recent years on American Journal of Epidemiology outlined. PMID- 9208517 TI - [Detection of poliovirus in fecal specimens by polymerase chain reaction]. AB - Poliovirus-RNA was detected in 11 clinical fecal specimens with AFP out of 19 samples. These comparative results between NT and Mcab identifications showed a coincidence rate of 90.90%-100%. Our experiment suggested that PCR a simple, sensitive, identification type and could be used for intratypic-differentiation of poliovirus strains. It can also be applied for the detection of poliovirus contamination in the environment. PMID- 9208518 TI - [Economic burden of poliomyelitis]. AB - Economic burden that caused by paralyzed and fatal poliomyelitis was described to provide basic data for evaluation. The study showed: the average costs for treatment, recovery and rehabilitation were 1612.6 yuan, 921.9 yuan and 1112.2 yuan per case, respectively. The total economic loss due to all new victims and handicapped cases of polio was 1.99 billion yuan in 1993 in China. PMID- 9208519 TI - [Role of Leptotrombidium (L.) scutellare in transmission of human diseases]. AB - A series of cases of hemorrhagic fever with renal syndrome (HFRS) in Shaanxi province and autumn-type tsutsugamushi disease in Jiangsu province were studied during 1988 to 1990 and 1986 to 1993 to explore the role of Leptotrombidium (L.) scutellare in transmission of these diseases. Results showed that L. (L.) scutellare is a dominant strain of mites inhabiting in rats and its seasonal fluctuation conformed to the seasonal distribution of these diseases. L. (L.) scutellare could naturally infect with HFRS virus and Rickettsia tsutsugamushi and transmit them by stinging and transovarial route. It indicated that L. (L.) scutellare could be a vector in transmission of HFRS and autumntype tsutsugamushi disease. PMID- 9208521 TI - [Micronucleus effect induced by Helicobacter pylori on human gastric mucous epithelial membrane]. AB - A large number of data from epidemiologic and statistical studies showed there was a relationship between infection of Helicobacter pylori (Hp) and gastric carcinogenesis, but its mechanism was unclear. Micronucleus test in vitro, one of cytogenotoxic studies, was carried out to study mutagenicity of Hp. Results showed micronucleus formation in gastric mucous epithelial membrane cell GES-1 treated with ultra-sonicated Hp for 72 hours was significantly greater than in the negative controls. There existed a dose-response relationship between them when concentration of Hp reached a range of 0.3-1.7 mg/L. As concentration of Hp reached 1.7 mg/L, percentage of micronucleus formation was five times higher than that of controls. When concentration of Hp increased to 4.3 to 10.7 mg/L, micronucleus formation did not increase but it was still over two times higher than that in controls. It suggests that there exists a direct cytogenotoxic effect of Hp on human gastric mucous epithelial membrane. PMID- 9208520 TI - [A study on transmission of bancroftian filariasis in Tancheng county, South Shandong province]. AB - A longitudinal study was conducted to explore transmission dynamics and epidemic trend of bancroftian filariasis in Huayuan Village, Shengli Township, Tancheng County, Shandong Province, a previous highly endemic area of it, from 1984, the fifth year after its preliminary eradication, to 1993, without any control measures being taken. Results showed proportions of residents with microfilaremia decreased to 0.12% in 1993 from 0.56% before the study. Eight of the nine microfilaria carriers before the study became negative spontaneously. Six new microfilaria carriers were detected during the study period, but five of them converted negative in three or four years. Negative conversion rate of blood IgG4 in microfilaremia cases was 88.89% by the end of the study. Natural infection rate in mosquitoes dropped year by year, with 0.21%, 0.19% and 0.06% for 1984, 1985 and 1986, respectively, and no filarial larvae was found in them in 1993 and natural transmission potential dropped from 3.47 to zero. It suggested the threshold for transmission of filariasis was not reached and the transmission was close to the end, with the rates of mosquito biting of 24.1-52.5 and 13.5-21.0 times per person, per night, for those using or non-using mosquito net, respectively. PMID- 9208522 TI - [Studies on relationship between toxicity of trichothecene toxin T-2 and its structure]. AB - Toxic action of mycotoxin T-2 and its metabolite T-2 tetraol and deepoxy T-2; tetraol was studied by cytotoxicity and animal toxicity tests to explore the relationship between toxin T-2 and its structure. Restults showed that proliferation of LLC-PK1 cells and synthesis of DNA could be inhibited by both toxin T-2 and T-2 tetraol. Toxicity of toxin T-2 was 100 times greater than that of T-2 tetraol. There was no obvious toxicity to the proliferation of LLC-PK1 cells and synthesis of DNA in a dose of 10 mg/L deepoxy T-2 tetraol. But, toxin T 2 caused obvious damage in heart muscle and articular cartilage of chicken embryos, and T-2 tetraol and deepoxy T-2 tetraol in heart muscle, but not in articular cartilage. T-2 tetraol produced by hydrolysis of toxin T-2 had toxicity to certain extent, but its strength was significantly less than that of the latter. It suggests that the epoxide group in toxin T-2 played a determinant role for its toxicity. If epoxy cycle of the basic nucleus of the molecule is open, its toxicity will change significantly. PMID- 9208523 TI - [A survey of moniliformin in rice and corn sampled from the areas with and without Keshan disease]. AB - Moniliformin (MF) was determined quantitatively for 123 samples of rice and corn collected from the areas with Keshan disease in Sichuan and Yunnan provinces and the areas without Keshan disease in Beijing using thin layer chromatography (TLC) and high performance liquid chromatography (HPLC). Results showed MF was found in 8.8% of the 83 rice samples from the areas with the disease, ranging from 73.5 to 265.3 micrograms/kg, and in 2.5% of the 40 sample from the areas without the disease with an average of 179.5 micrograms/kg. MF was found in 81.4% of the 43 corn samples from the areas with the disease, ranging from 52.3 to 1116.0 micrograms/kg, and in 9.7% of the 61 samples from the areas without the disease ranging from 73.3 to 1082.2 micrograms/kg. There was significant difference between positive proportions for MF in rice and corn from the diseased and non diseased areas, but no significant difference between their average amount of MF. Differences in contamination of MF between rice and corn was much greater than that between different areas. PMID- 9208524 TI - [Urine level of trans, trans-muconic acid used as an index of internal dose of exposure to benzene]. AB - Urine trans, trans-muconic acid (TTMA) in population exposed to benzene was determined with a high performance liquid chromatography-ultra violet detection system. Result showed urine TTMA increased significantly with increasing of air benzene levels, showing a good correlationship between them (r = 0.940, P < 0.05). Even in relatively low air benzene levels (with an average of 2.46 mg/m3), urine TTMA in exposed group was higher, as compared with that in controls, and their difference reached statistically significant level. It suggests that urine TTMA level can be used as a sensitive and specific indicator for biological monitoring, especially for low exposure to benzene. There was no significant difference in urine TTMA between males and females exposed. Urine TTMA formation can be inhibited by toluene, and confounding of toluene can be roughly adjusted by analysis of covariance. PMID- 9208525 TI - [Seven-year follow-up in pupils immunized with hepatitis B vaccine and its revaccination]. AB - In order to evaluate immune persistence of plasma-derived hepatitis B vaccine (HBV), 344 children in a primary school were followed up for seven years by detection of serum HBsAg, anti-HBc and anti-HBs with radioimmunoassay (RIA) using Abbott reagent kits. In the seventh year after three-dose HBV immunization, level of anti-HBs in 55.4%-75.0% of the vaccinee was greater than 10 IU/L, with a geometric mean titer (GMT) of 20.9-65.3 IU/L. The higher the level of anti-HBs, the longer the duration of its decrease from more than to less than 10 IU/L. Seven years after immunization, 66 children with varied serum levels of anti-HBs were received a fourth dose of 10 micrograms HBV, and one month after revaccination, their levels of anti-HBs increased by 6.3-57.7 times. It suggests revaccination of HBV is not necessary for children vaccinated within seven years. PMID- 9208526 TI - [A genetic epidemiological study on lung cancer]. AB - Segregation ratio, heritability and relative risk of genetic susceptibility were estimated for 355 families of lung cancer in matched pair with genetic epidemiological methods. Results showed that segregation ratio for lung cancer was 0.09-0.12 (95% confidence interval, CI) and heritability of lung cancer was (40.58 +/- 4.01)% and (27.58 +/- 4.76)% for smokers and non-smokers, respectively. Relative risks of genetic susceptibility to lung cancer were 4.73 (95% CI 3.90-5.74) and 2.61 (95% CI 2.18-3.13) in their first and second degree relatives, respectively, after adjustment of smoking with a logistic regression model. It was also found that there was an interaction between smoking habit and genetic background of lung cancer. Thus, it is believed that genetic background is one of the multifactorial causes in lung cancer. PMID- 9208527 TI - [An analysis of CMV infection in 115 cases with viral hepatitis]. AB - To study prevalence of cytomegalovirus (CMV) infection in patients with viral hepatitis and its clinical characteristics, serum anti-CMV-IgM was detected in 6411 hospitalized hepatitis cases, and clinical symptoms, signs and liver function in 115 cases with CMV infection were compared with 192 cases of non-CMV infection. Results showed a CMV infection rate of 1.79% in them with an average age of 33.6 years, a sex ratio of 2.13, and dual superinfection with CMV and hepatitis accounting for 44.74%, and triple and quadruple superinfection for 47.37% and 7.89%, respectively. Proportion of those with fever, digestive symptoms, hepatomegaly, changes of gallbladder in ultra sound scan, rising activities of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and gamma-glutamyltransferase (gamma-GT) were more, and duration of hospitalization longer in the cases with CMV infection. It suggests that CMV can be found in the cases with viral hepatitis, most in a form of dual or multiple infection, and it can aggravate hepatitis. PMID- 9208528 TI - [Studies on strategies for changing food composition of residents based on their nutritional status]. AB - Investigations of resident food consumption and evaluations of their dietary nutrition were carried out to correctly lead to the adjustment of food composition, promotion of coordinating development of food consumption and production, and continuous improvement of people's nutritional level and health status. Results showed that residents consumed less protein of high quality, more fat in the urban area, insufficient some trace element and more salt in Hebei province. Based on these facts, it is recommended that to consume vegetable food as major dietary composition with a variety of grains and vegetables, to increase adequately animal food and legumes, to decrease salt intake, and to lower fat intake in the urban areas be the strategic goals for the food composition changes. Measures to implement these goals are discussed in the paper, also. PMID- 9208529 TI - [Investigation of health status in workers exposed to low-level benzene]. AB - In order to know the health status in workers exposed to low-level benzene, 323 air samples were collected from five workshops of the Corporation, where low level benzene was emitted from aromatic hydrocarbon devices. Results showed air benzene levels ranged from 0.05-29.25 mg/m3, with an average of 4.57 +/- 6.16 mg/m3, and the highest in the benzene-producing workshop. Four hundred and thirty seven workers closely-exposed to benzene in five workshops and 150 controls were selected and their peripheral white blood cell count and micronucleus of lymphocytes were determined. Result showed that 1. peripheral white blood cell count of the exposed workers was within the normal range, but obviously lower than that of the controls; 2. proportion of lymphocytic micronucleus was higher in exposed workers than that in unexposed ones, and there was significant difference in it between workers in benzene-producing workshop than that in controls; 3. white blood cell count lowered and proportion of lymphocytic micronucleus increased in the workers, with the increase of air benzene level in the five workshops. All these demonstrate that low-level benzene is harmful to the exposed workers, especially to their genetic materials. PMID- 9208530 TI - [Advances in the research on seroepidemiology of human herpesvirus-6 infections]. PMID- 9208531 TI - [Clinical and experimental study on treatment of rotaviral enteritis with qiwei baizhu powder]. AB - Sixty cases of rotaviral enteritis treated with Qiwei Baizhu Powder (QWBZP) revealed a better efficacy than that treated with Oral Rehydration solution (ORS, chi 2 = 6.07, P < 0.05). The content of Na+ and glucose as well as number of patients with positive human rotavirus (HRV) antigen in faeces in QWBZP group were less than that in ORS group (chi 2 = 18.09, P < 0.05). In experimental study, QWBZP was found to be effective in treating HRV enteritis of newborn NIH mice in vivo, as compared with the control groups, the mortality of mice was decreased by 73.3%, the content of Na+ and glucose as well as number of mice with positive HRV antigen in faeces was markedly reduced, the pathological changes of intestine such as the damage of small intestinal mucosa and the exfoliation of intestinal villi were also obviously alleviated. PMID- 9208533 TI - [Effect of acupuncture on immunomodulation in patients with malignant tumors]. AB - In order to investigate the role of acupuncture in the regulation of cellular immune function, the changes of T lymphocyte subsets (CD3+, CD4+, CD8+), soluble interleukin-2 receptor (SIL-2 R) and beta-endorphine (beta-EP) in the peripheral blood of patients with malignant tumors before and after acupuncture were observed with double blind method. Forty patients were divided randomly into two groups, 20 for each. One group treated with acupuncture and the other one for control. Results showed that acupuncture has the effect of enhancing the cellular immunity of patient with malignant tumor. Acupuncture treatment could increase the percentage of T lymphocyte subsets CD3+, CD4+ and the CD4+/CD8+ ratio (P < 0.01) and the level of beta-EP, as well as decrease the level of SIL-2 R (P < 0.01). The correlation analysis of the three criteria showed there was a positive correlation between beta-EP and T lymphocyte subsets and a negative correlation between beta-EP and SIL-2 R, there was also a negative correlation between T lymphocyte subsets and SIL-2 R. Based on these results, a discussion on the acupuncture immunomodulation network was conducted in this article in order to explore the possible mechanism of acupuncture on immunomodulation. PMID- 9208532 TI - [A study on treatment of lung cancer by combined therapy of traditional Chinese medicine and chemotherapy]. AB - From May 1992 to May 1995, 102 patients with lung cancer were divided randomly into two groups: group A, 48 patients, treated with combined therapy of TCM and chemotherapy, group B, 54 patients treated with chemotherapy alone. The protocol of chemotherapy used in the two groups was similar. The Chinese medicines were given according mainly to the Syndrome Differentiation. Four types were found in the group A: the Lung and Spleen Qi Deficiency type, the Lung Heat and Phlegm Dampness type, the Lung-Yin and Stomach-Yin Deficiency type and the Qi stagnation and blood stasis type. The total effective rate according to the WHO standard was as follows: that of group A and group B was 52.1% and 35.1%; the 1 year survival rate of them was 69.4% and 66.7%, the 2 year survival rate was 56.2% and 15.8% and the median survival time were 13 months and 9 months, respectively. These results suggest that the elevation of median survival time and 2 year survival rate of group A might be closely related with the therapeutic effect of TCM, but the combined therapy of TCM and chemotherapy did not improve the therapeutic efficacy significantly. PMID- 9208534 TI - [Effect of shengmaisan on serum lipid peroxidation in acute viral myocarditis]. AB - The effect of Shengmaisan (SMS) on 62 acute viral myocarditis patients and its peroxidation damage was studied. The results revealed that the activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in blood were decreased and the content of malondialdehyde (MDA) in plasma was increased in acute viral myocarditis patients in comparison with the healthy controls (P < 0.001). 62 acute viral myocarditis patients were divided into two groups: SMS group and placebo group. After treatment, both SOD and GSH-Px activities were increased and the level of MDA decreased (P < 0.001) in SMS group, while those in placebo group were not changed (P < 0.05). The results suggested that the myocardial damage of viral myocarditis is closely related with lipid peroxidation SMS acts as an effective free radical scavenger and anti-lipid peroxidation drug. SMS could prevent the damage of myocardia and might be taken as one of the effective therapeutic methods in treatment of acute viral myocarditis. PMID- 9208535 TI - [Influence of qingdai compound capsule (QDCC) on the expression of c-myc in psoriatic keratinocytes]. AB - In this immunohistochemical study the authors detected the expression of c-myc in keratinocytes (ECK) from 30 psoriatic patients before and after QDCC treatment and 12 normal subjects for control. The results showed that the ECK of patients before treatment was significantly higher than that in normal control, and its level was correlated with histopathological changes. After treatment the ECK was normalized. It is believed that QDCC could decrease the EKC in psoriatic patient and this effect probably was mediated by inhibiting c-myc expression. This study provided an experimental evidence for the application of QDCC in treating psoriasis. PMID- 9208536 TI - [Clinical study on anti-senility effect of kuangquan 851 oral liquor type R and type Y]. AB - The anti-senility effects of Kuangquan 851 Oral Liquor type R and type Y were studied and compared with Qing Chun Bao Oral Liquor as control in 303 subjects of geratic period or presenium with senile syndromes of Kidney or Spleen Deficiency. The results indicated that both type R and Y could improve the Kidney and Spleen Deficiency. The effective rates of type R and Y were 96.6% and 92.3% respectively and were much better than that of the control group. The laboratory findings before and after treatment revealed that effects of Kuangquan 851 Oral Liquor were as follows: increasing antioxidation and clearing away free-radicals; promoting collagen metabolism; enhancing the function of T lymphocytes; improving pulmonary, cardiovasular, brain function; raising male serum testosterone and estradiol levels. The two types were similar in effect. PMID- 9208537 TI - [Influence of different combination of mental activity and respiratory cycle on heart rate variability]. AB - By means of spectral analysis of P-R interval, different characteristics of heart rate variability in different form of respiratory exercise was observed. The results of observation on 32 volunteers showed that mental activity that affected respiration can influence the function of autonomic nerve system in a different way. When the mind was concentrated at the inspiration, the function of autonomic nerve system was kept in balance, and both the sympathetic and the vagal activities enhanced significantly and while mind concentrated at the expiration could induce a reduction of vagal activity so as to produce marked change in the sympathovagal balance. PMID- 9208538 TI - [Effect of concentrated xuefu zhuyu pill on proliferation of vascular smooth muscle cells in experimental atheriosclerosis rabbits observed by serologic pharmacological test]. AB - Two in vitro experiments were performed for comparative study to evaluate the effect of concentrated Xuefu Zhuyu pill (CXZP), a Chinese traditional herbal medicinal mixture, on the proliferation of vascular smooth muscle cells (SMCs) of experimental atherosclerosis rabbits by serologic pharmacological test. The first experiment approach adopted the addition of the rabbit's serum to the culture medium while in the second experiment the CXZP was added to the medium directly. Results showed that both the serum with CXZP and the CXZP extract itself could inhibit the proliferation of SMC significantly and dose-dependently, and the inhibitory action of the latter was more potent than the former one. It suggested that CXZP could inhibit the proliferation of SMC both in vivo and in vitro, the decrease of effectiveness in vivo might be due to the non-absorption of some active ingredient of CXZP or due to metabolic inactivation. PMID- 9208539 TI - [Mechanism of injury of intestine, liver and lung in acute cholangitis of severe type model and the protective effect of huoxue qingjieling]. AB - Lung capillary permeability was measured with 125I labelled bovine serum albumin, the progression of hepato-cellular injury was quantitatively assessed using atriphenyltetrazolium chloride assay, injury of small intestinal mucosa was determined by the changes of diamine oxidase (DAO) level, content of oxygen free radicals (OFR) was measured by electron spin resonance spectrometer and levels of phospholipase A2(PLA2) of intestine, hepatocellular viability were observed by bioassay methods respectively. The results showed that function of lung, liver and intestine was seriously injuried in ACST rats. These injuries demonstrated by increase of capillary permeability and decrease of hepatocellular viability as well as DAO in intestinal mucosa (P < 0.01) in comparing with control, while the levels of OFR and PLA2 in the above-mentioned 3 organs were elevated simultaneously (P < 0.01). As for the comparison of efficacy of different treatment, Huoxue Qingjieling was the best one, the bile duct decompression or ampicillin showed little benefit on injury protection even aggravated in some tissues, they displayed a close relationship with their influence on the OFR and PLA2. Besides, the levels of OFR or PLA2 in various tissues were different from each other, therefore, the serum levels of OFR or PLA2 can not reflect the real levels occurred in organ or tissue level. PMID- 9208540 TI - [Effect of shashen maidong decoction on gastric motility in vivo]. AB - Effect of Shashen Maidong Decoction (SSMDD) on the gastric motility of mice and rats was observed in vivo. Results showed that the gastric phenol red excretion rate of mice could be reduced markedly by SSMDD given in various dosages (23 g.kg 1.d-1, 9 g.kg-1.d-1 or 23 g.kg-1.d-1 for 5 days successively), P < 0.05. By administration of 9 g.kg-1.d-1 or 23 g.kg-1.d-1 could antagonize the acceleration of gastric emptying induced by neostigmine in mice (P < 0.05, P < 0.01). 7 g/kg of SSMDD by gastrogavage could inhibit the frequency and amplitude of contraction of fundic longitudinal muscle using strain gauge transducer in rats, but the effect was not obvious on that of antral circular muscle. By water-ballon method, it was observed that SSMDD 3.5 g/kg or 7 g/kg intraduodenal perfusion could slow down the frequency and decrease the amplitude of gastric peristalsis, 7 g/kg could inhibit significantly the gastric hypermotility induced by subcutaneous injection of indomethacin (40 mg/kg), but had no obvious effect on the gastric hypermotility induced by intramuscular injection of reserpine (0.4 mg.kg-1.d-1 x 4 d). PMID- 9208541 TI - [Regulatory effect of lingqi anshen liquor on erythrocyte immune function and antioxidation in immunosuppressed mice]. AB - The regulatory effect of Lingqi Anshen Liquor (LQASL) on erythrocyte immune function and antioxidation in cyclophosphamide induced immunosuppressed model mice was observed. After given LQASL for 7 days, the results showed that it could antagonize the inhibitory action of cyclophosphamide significantly, demonstrated by raising the level of superoxide dismutase activity, erythrocyte immune adhesive enhance factor and erythrocyte C3b receptor rosette forming rate, lowering the serum malondialdehyde, erythrocyte adhesive inhibitory factor and erythrocyte immunocomplex rosette forming rate. It suggested that LQASL can regulate the erythrocyte immune function and antagonize the injury of oxygen free radical in immunosuppressed mice. PMID- 9208542 TI - [Mechanism of the treatment of pigmentary degeneration of retina with traditional Chinese medicine]. PMID- 9208543 TI - [Review and prospect of making blood-stasis animal model]. PMID- 9208544 TI - [Recent progresses in pharmacological and clinical studies of paeonol]. PMID- 9208545 TI - [Radix Sophorae flavescentis quality by chemical pattern recognition]. AB - The PCA method and NLM technology were used to study on quality of 40 Radix Sophorae Flavescentis samples collected from different regions of China. The results showed were unanimous with that of traditional appraisal. PMID- 9208547 TI - [Commodity identification of semen Plantaginis and herba Plantaginis]. AB - The Semen Plantaginis from 29 provinces and regions and the Herba Plantaginis from 18 provinces and regions in our country were identified, and the main commodities of these two crude drugs are described in this paper. The identification may serve as a scientific basis for expanding new drug resources. PMID- 9208546 TI - [Dynamic assay of oleanolic acid in Aralia taibaiensis Z. Z. Wang et H. C. Zheng]. AB - The dynamic assay on oleanolic acid (OA) in Aralia taibaiensis shows that the plant has a higher OA level. The level in root cortex is 4.74%-10.81% and peak stages are mid-May to late June and late August to mid-September the level in stem barks is 3.59%-12.06% and peak period is between mid-May and late July, the level in leaves increases throughout the life time ranging from 2.41% to 6.66% and peak level is 6.66% around mid-September. PMID- 9208548 TI - [Correlative study on the storage time and quality of rhizoma Dryopteris crassirhizomae]. AB - The contents of dryocrassin in Rhizoma Dryopteris Crassirhizomae were determined by TLC-scanning in a period of two years of storage. Phloroglucinol content was calculated by area normalization method. The results show that in contrast with the fresh crude drugs of Rhizoma Dryopteris Crassirhizomae, the contents of dryocrassin in the crude drugs of one year and two years of storage have reduced by 22% and 27% respectively, and phloroglucinol contents have reduced by 2% and 8% respectively. PMID- 9208549 TI - [Effects of sand and hormone treatment on seed germination of Bupleurum chinese DC. and B. falcatum L]. AB - The seeds of Bupleurum chinese and B. falcatum were treated first by GA3 or 6-BA in different concentrations and then by sand. The best method for promoting their germination has been developed. PMID- 9208550 TI - [Acute toxicity and anti-inflammatory effect of processed products of gamboge]. AB - The acute toxicity and anti-inflammatory effect of processed and raw Gamboge were compared. The results showed that the acute toxicity of processed Gamboge was less than that of the raw one, while both had significant anti-inflammatory effect alike. Among the differently processed products, those boiled with lotus leaf juice and steamed by high pressure were better. PMID- 9208551 TI - [Comparative research on the constituents of the volatile oil in the rhizome of Cibotium barometz (L.) J. Sm. and its processed products]. AB - Volatile oil was extracted from the rhizome of Cibotium barometz for the first time. Its content was determined to be 83.3 micrograms/g, decreasing significantly after the drug was processed. Twelve constituents from the volatile oil were identified by method of GC-MS. The main constituents were organic acids. The contents of palmitic acid and linoleic acid were the highest. After Cibotium barometz was processed, some constituents were found decreasing and some increasing. PMID- 9208552 TI - [Preparation of huichun zhibao oral liquid (HZOL)]. AB - The formulation, preparing method, quality standards, pharmacodynamic experinents and toxicological experiments of HZOL are described. The content of icariin, which is and effective component of epimedium herb as well as one of the main ingredients in HZOL, was successfully determined through HPLC, with an average recovery of 98.98% and RSD = 1.53%. PMID- 9208553 TI - [Experimental studies on the quality control of xiaoluo pills]. AB - Microscopic and physico-chemical methods have been used in the qualitative study of Xiaoluo Pills. The contents of peimine, peiminine and calcium carbonate in these pills have been determined. The proposed method can be used to control the quality of Xiaoluo Pills. PMID- 9208554 TI - [Chemical constituents of Taxus yunnanensis Cheng et L. K. Fu]. AB - Five compounds were isolated from the bark of Taxus yunnanensis. Their structures were identified on the basis of spectral analysis and physico-chemical constants as taxol C-7-xylose, taxol B, 10-deacetyl baccatin III, taxusin and 2R,3R catechin. PMID- 9208555 TI - [Alkaloids in the flowers of Sophora viciifolia Hance]. AB - Six alkaloids have been isolated from the flowers of Sophora viciifolia for the first time. Based on of physico-chemical methods and spectroscopic analysis, their structures have been identified as oxymatrine, oxysophocarpine, sophocarpine, matrine, sophoramine and sophoridine. PMID- 9208556 TI - [Chemical constituents of Gossampinus malabarica (L.) Merr. (II)]. AB - Four chemical compounds isolated from the root of Gossampinus malabarica were identified as daucosterol, oleanolic acid, hesperidin and potassium nitrate by physico-chemical constants and spectroscopic analysis. PMID- 9208557 TI - [Constituents of essential oil of imported myrrh and gum opoponax]. AB - The constitutents of essential oil in two kinds of Myrrha were analyzed by GC-MS. Fifteen compounds in Myrrh and thirty-three compounds in Gum opoponax were identified with their percent contents given. The main constituent of Myrrh is furanoeudesma-1,3-diene, and the main constituent of Gum opoponax is beta-trans ocimene. PMID- 9208558 TI - [Determination of genkwanin in flos Genkwa by HPLC]. AB - In this paper, the method for determining genkwanin in Flos Genkwa was established by HPLC. Detected at 332nm on a Lichrosorb 5 RP-18 column with a mobile phase of methanol-water-acetic acid (65:35:5), the content of genkwanin in Flos Genkwa was determined to be 0.16%. The recovery rate was 95.46% and RSD 1.15%. PMID- 9208559 TI - [Pharmacological action of siwei shaoyao decoction]. AB - The Siwei Shaoyao Decoction possesses a marked effect on the alleviation of trigeminal neuralgia in rats caused by penicillin G potassium injection. As shown from the hot-plate test, it also has an obvious analgesic effect on mice. To some extent, the decoction has a significant anti-inflammatory effect on the acute edema in hind paws of rats and the effect is believed to be related to the reduction of capillary permeability. PMID- 9208560 TI - [Inhibitory effect of radix et rhizoma Rhei on the production of lipid peroxides (LPO) in mice liver]. AB - The experiment has shown that the aqueous extract and alcohol extract of Radix et Rhizoma Rhei can markedly inhibit the production of LPO in mice liver in vitro, and the aqueous extract also restrains the LPO increase in mice liver-caused by adriamycin or D-galactose. PMID- 9208561 TI - [Antimicrobial properties of Flos Lonicerae against oral pathogens]. AB - The antimicrobial effect of Flos Lonicerae on oral pathogens was studied. The results showed that 73.9% of the tested pathogens were inhibited at a concentration below 6.25mg/ml. Streptococci mutants, actinomyces viscosus and bacteroides melaninogenicus were comparatively more sensitive to Flos Lonicerae. PMID- 9208562 TI - [Effect of Chinese herbs on the circadian rhythm of body temperature and heart rate in rabbits with hypothyroidism (yang deficiency)]. AB - The rectal temperature (Tr) and heart rate (HR) of three groups of New Zealand white rabbits, i. e. thyroidectomized (G1), thyroidectomized and treated with Chinese herbs (a decoction composed of tonifying kidney-yang herbs) (G2) and pseudothyroidectomized for control (G3), were observed dynamically for 26 hours. The results indicated that the Tr and HR in G1 were decreased, and the circadian rhythms of the Tr and HR disordered significantly, while these changes in G2 were corrected obviously. PMID- 9208564 TI - [Detection of FMR-1 gene expression by RT-PCR]. AB - Fragile X syndrome [FRA(X)] as the most common form of inherited mental retardation in man has an incidence of one per 1250 and is associated with a fragile site at Xq27.3. A gene was identified at the fragile X locus and was designated Fragile X Mental Retardation-1 (FMR-1). FRA(X) resulted from expansion of (CGG)n trinucleotide repeat in 5' untranslated region of the human FMR-1 gene, and was associated with abnormal methylation of a CpG island 250 bp proximal to this (CGG)n repeat. Males with typical FRA(X) showed repression of FMR-1 transcription and absence of FMR-1 protein, which was believed to contribute to the fragile X phenotype. FMR-1 mRNA extracted from leukocytes in normal and clinically suspected males were detected by RT-PCR. The methylation status and CGG expansion were also studied by PCR and Southern blot. Two of 10 clinically suspected males were found devoid of FMR-1 expression and accompanied with hypermethylation of the CpG island and CGG trinucleotide repeat expansion. PMID- 9208563 TI - [The mRNA expression of CD2, CD4 and CD8 in peripheral blood lymphocytes from the patients with acute lymphoblastic leukemia and non-Hodgkin's lymphoma]. AB - The paper for the first time reports the detection of the levels of CD2, CD4 and CD8 mRNA expression in peripheral blood lymphocytes from 9 cases with T-acute lymphoblastic leukemia (ALL) and 10 cases with T-non-Hodgkin's lymphoma (NHL) by using RNA dot blotting technique. It was found that the levels of CD2 mRNA in the two groups of patients were significantly higher than those in the normals. The ratios of the CD4 to CD8 mRNA were also increased in patients with NHL. The significance of this study was discussed. PMID- 9208565 TI - [Effect of cholecystokinin on prolactin release and its action mechanism in the rats]. AB - The effect of cholecystokinin octapeptide (CCK-8) on the release of prolactin (PRL) in male rats were studied in vivo and in vitro. CCK-8 at the concentrations (microgram) of 0.05 and 0.5 was injected into the third cerebral ventricle (3rd, V. I) of conscious rats, outfitted with chronic 3rd. V. and jugular cannulae, a significant increase in resting secretion and restraint stress-induced release of PRL were observed. The effects of CCK-8 at the concentration of 0.05 microgram were stronger than those of 0.5 microgram. To determine if CCK-8 would exert any direct action on anterior pituitary, CCK-8 of 0.05, 0.5, 1.00 microgram were added to the medium of dispersed anterior pituitary cell, and caused dose dependent increase of PRL secretion. To study a mechanism of intracellular signal transduction in the action of CCK-8, the levels of cAMP and [Ca2+] in the medium were measured. Intracellular Ca2+ concentration of disperse anterior pituitary cell was significantly elevated by CCK-8 (2 x 10(-4) mol/L), but CCK-8 (10(-8) 10(-6) mol/L) did not change intracellular cAMP content. The results indicate that CCK-8 stimulate prolactin release at both sites of hypothalamic and anterior pituitary and the mechanism of stimulating effects of CCK-8 might be mediated by [Ca2+] but not cAMP. PMID- 9208566 TI - [The effect of intra-aortic balloon pumping in case of left main coronary artery stenosis]. AB - In order to improve perfusion of coronary artery in case of coronary artery stenosis, effect of intra-aortic balloon pumping (IABP) on coronary blood flow (CBF) was investigated. To this end hemodynamic parameters of heart functions with and without stenosis were analysed under IABP. Hemodynamic effect of IABP was obviously influenced by the coronary artery stenosis. PMID- 9208567 TI - [The study of the relationship between the granule size of immunoliposomes and the conjugate ratio of F(ab')2 of McAb HI30 (CD45)]. AB - The immunoliposomes were prepared by conjugating the F(ab')2 of monoclonal antibodies HI30(CD45) labelled by 125I with the liposomes labelled by 131I, and then were separated by a sephacryl S-300 column. The chromatography showed that the liposomes prepared by ultrasonic method were not homogeneous, and the liposomes of smaller granule size had a higher conjugate ratio. It was also found that the fragment F(ab')2 of McAb conjugated to liposomes could enlarge their granule size. PMID- 9208568 TI - [Species-specific monoclonal antibodies against the major outer membrane protein (MOMP) of Chlamydia trachomatis]. AB - The synthesized one quarter N-terminal MOMP of C. trachomatis was used for primary immunization of three male BALB/c mice (8 weeks of age), and the boost with C. trachomatis L1/440/Bu elementary bodies (EBs) was followed on day 14. Spleen cells from one mouse with good response of immunization were fused with murine myeloma NS-1 cells on day 24. The hybrid cell suspension was seeded into the wells of 96-well microtest plates which contained macrophage feeder layers. Anti-chlamydial antibodies in culture fluids were screened by ELISA with 1/4 MOMP & L1 EBs coated 96-well trays. Positive wells were cloned by limiting dilution. Four clones which secreted immunoglobulin G1 & G2a class were obtained after elimination of those clones that produced antibodies to C. psittaci strain EAE, C. pneumoniae strain ATCC VR1310 and uninfected BGMK cells. In micro-IF test, we found that the all four clones of MAbs reacted with our laboratory prepared L1, L2, A, B, C, E EBs, L2 tissue culture inclusions, as well as the EBs of all 15 standard serovars of C. trachomatis. The titers of their ascites were more than 1:12,800 in micro-IF test. It was shown that the four clones of MAbs reacted predominantly with 40,000 MOMP of C. trachomatis L1 in Western blot. PMID- 9208569 TI - [Study on animal models of immune mediate motoneuron disease]. AB - Swine motoneurons (SMN) were isolated from fresh spinal cords of pigs. Homogenates of these SMN or fresh anterior horns of pigs (SAH) as immunogens were inoculated to guinea pigs or Lewis rats and Wistar rats respectively. The impairments of motion were observed in the immunized guinea pig four months after the fifth inoculation of SMN, in the immunized Lewis rats after the first or second inoculation of SAH and in the Wistar rat after the third inoculation of SAH respectively. Degeneration and loss of motoneurons in the spinal cords of these symptomatic animals were found histologically. Antibodies against motoneurons of guinea pig and rat can be detected in sera of these symptomatic animals with immunocytochemical method respectively. In control guinea pigs, Lewis rats and Wistar rats there were no symptom, and did not found degeneration of motoneurons in the spinal cords of these control animals. Antibodies against motoneurons can not be detected in the sera of these control animals. The results indicated immune mediate guinea pig or rat models for MND can be established with pure SMN or impure SMN as immunogens. It was shown the conservative homology between antigenic structures of lower motoneurons in pigs and guinea pigs or rats. The pathogenesis in immunized animals with SAH is faster than that in immunized animals with SMN at least in terms of the appearance of symptoms. The Lewis rats produced symptoms first and the incidence ratio of symptomatic animals in Lewis rats was the highest. The immunized Wistar rats produced symptoms a little bit slower. PMID- 9208570 TI - [Neuro-specific enolase in acute ischemic stroke and related dementia patients]. AB - Neuron-specific enolase (NSE) levels of cerebrospinal fluid (CSF) were measured in 39 patients with ischemic stroke and 15 controls. There was a significant increase of CSF NSE in patients with acute ischemic stroke as compared with the controls. The altered CSF NSE levels were well correlated with the infarct size in CT scan. The CSF NSE levels were higher in 6-patients who were diagnosed as multi-infarct dementia (MID) after 6-month follow-up than in 22 non-MID patients of this series. Our research supports the view that CSF NSE can be a useful biochemical marker for brain ischemia. The importance of CSF NSE for dementia related to ischemic stroke is worth further studying. PMID- 9208571 TI - [Surgical treatment of spinal tumors: a report of 30 cases]. AB - With the advance of spinal surgery in the last decade, surgical treatment of spinal tumors has been no longer limited to sole laminectomy. The principles of surgical treatment of spinal tumors include: (1) anterior approach for the anterior lesion and posterior approach for the posterior lesion, (2) combined anterior and posterior approach for extensive lesions and (3) internal fixation for spinal stability. 30 cases of spinal tumors were treated on the basis of the above guiding principles and 83.3% (25 of 30 cases) showed excellent or good outcomes. PMID- 9208572 TI - [A prospective randomized study of parenteral and enteral nutrition in postoperative patients]. AB - To evaluate the metabolic effects of balanced enteral nutrition (EN) and standard parenteral nutrition (PN, without glutamine) in post-operative patients, we performed a prospective randomized trial in 24 patients after gastrectomy or semi colectomy. All patients had normal liver, kidney, heart and lung functions. patients of the enteral nutrition group received liquid Ensure (Abbott-Ross) while patients of the parenteral nutrition group received standard parenteral nutrition. Parenteral nutrition solutions consist of amino acids, fat emulsions, glucose, electrolytes, vitamins and trace elements. Subjects of the two groups received iso-nitrogenous and isocaloric nutritional regime for seven days (from POD+5 to POD+11). Weight loss in the parenteral nutrition group and the enteral nutrition group was 1.7 +/- 0.2 kg and 0.8 +/- 0.31 kg respectively. Cumulative nitrogen balance was -82 +/- 51 mg.kg-1/d in the parenteral nutrition group vs +70 +/- 7 mg.kg-1/d in the enteral nutrition group (P < 0.05). The results showed that balanced enteral nutrition had less weight loss and better cumulative nitrogen balance than standard parenteral nutrition in this protocol. More patients study will be needed. PMID- 9208573 TI - [Ultrasonic diagnosis of malignant trophoblastic tumor (an analysis of additional 41 cases)]. AB - Forty one cases of malignant trophoblastic tumor which were confirmed by pathological examinations were reviewed. The diagnostic correspondence rate by sonography was 92.3% (38/41). The image patterns, types of tumor, differential diagnosis and diagnostic value of sonography were also explored in this paper. PMID- 9208574 TI - [Effect of the active components of Ranunculus ternatus Thunb. on the inductive production of tumor necrosis factors by macrophages]. AB - The authors set up a screening system for the detection of TNF inducers. By using this model 36 traditional Chinese medical herbs from 28 families were studied. Seven herbs from 7 families were found to be active in the TNF induction. Among them Ranunculus Ternatus (RT) is the best one. The active components of RT were isolated from the crude extract by solvent extraction and column chromatography. Eleven fractions were obtained. RT-A2, one of the active components, is a white crystal and its IR spectrum is similar to Standard Infrared Grating Spectra No. 28344K. Therefore, RT-A2 may be identified as a fatty acid (palmitic acid). Its optimum dose of inducing TNF is 50 micrograms/ml in vitro. TNF induced in the cell culture was demonstrated by monoclone antibody neutralizing test. PMID- 9208575 TI - [Acute cholangitis due to bile duct stones: clinical analysis of 85 cases]. AB - A retrospective review of 85 patients with acute calculous cholangitis who underwent emergency surgery (n = 85) was carried out. There were 28 men and 57 women and their mean age was 61.4 years. All patients had abdominal pain, 81% had chills and fever, 75% had clinical jaundice, and 28.2% were in shock. There were 48 severe acute cholangitis (group 1) and 37 acute cholangitis (group 2). The incidence of a positive bile culture in group 1 was 100% (28/28) and that of group 2 was 85% (17/20). The morbidity and mortality in group 1 was 43.8% and 20.8%, respectively. Five patients (13.5%) in group 2 had simple wound infection, and there was no severe complications or death. A significantly lower platelet count was documented in group 1 (86.5 +/- 40.7 x 10(9)/L) as compared with group 2 (159.9 +/- 63.9 x 10(9)/L) (P < 0.001). It is suggest that platelet count less than 90 x 10(9)/L at the time of admission or preoperation may serve as a guideline for identifying high-risk patient in early surgical intervention. PMID- 9208576 TI - [Manufacture of computer-based testing and analyzing system for measuring step through performance]. AB - The computer-Based Testing and Analysing System for measuring Step-Through (avoiding darkness) performance (CTASS) has been developed. The system is based on the infrared beams which transfer actions of mice inside the boxes into signals of voltage. The signals are fed into the computer through A/D converter to computer for software processing. The functions which have been realized include, but not limited to: signal sampling and data curve displaying; intelligent identification and elimination of jamming signal; auto-calculation of number of errors (No) in all channels, latent period (LP) of entering into the light chamber, total time of mice staying in the light chamber (safty place Tl) and dark chamber (error place Td); hardcopy of screen graphics and resulting output by various printers. The CATSS system was applied to test normal mice and the cognition-deficient mice induced by anisodine, a M-Cholinergic antagonist at a dose of 1-10 mg/kg i.p. The data collected by the system were compared and proved to be consistent with those acquired by manual labour. The system has produced refined and accurate information refined to a maximum extent and guaranteed scientifically sound experimental results. It is foreseable that introducing the computer into pharmacological researches will update and advance their methodology. PMID- 9208577 TI - [Gene therapy and adenovirus vectors]. PMID- 9208578 TI - [Sequencing of right terminal fragment of canine adenovirus type 1 vaccine strain DNA]. AB - Current studies show that adenoviruses are more advantageous as expression vectors than other viruses. Canine adenovirus type 1 (CAV-1) vaccine strain, CLL (cannaught laboratory limited), as vectors will play more important role for the prophylaxis of zoonosis than human adenoviruses. Studies on molecular biology of CAV-1 or CLL are necessary for construction of vectors of CLL. In this paper, the right terminal of CLL DNA cleaved by Hind II was sequenced. Authors found that it contained a 238 bp inverted terminal repeats (ITR). There are three copies of 40 bp short repeat sequences in the ITR. According to the DNA structural model of human adenovirus type 2, the 40 bp short repeats are the sites that are combined with nuclear factor 1 (NF-1) that is an initiation factor of DNA replication of virus, so they are essential structure of CLL DNA. PMID- 9208579 TI - [Secretion of endothelin-1 by cultured human normal adrenal cells]. AB - Endothelin (ET) is a vasoactive peptide produced by some other types of cells in addition to endothelial cells. The authors investigated into the possibility of ET-1 secretion by cultured human normal adrenal cells. Human normal adrenal cells were prepared by 2% collagenase digestion for 1.5 hours and cultured in DMEM supplemented with 10% FCS. Angiotensin I (10(-9)-10(-7) mol/L) was added to the experimental groups on the 7-9th days of cultivation. Mediums were collected after 24 hours and the levels of aldosterone, cortisol and ET-1 in the mediums were measured by RIA. Other than aldosterone and cortisol, ET-1 was detected in the cultured mediums of human normal adrenal cells. And the secretion of ET-1 was stimulated by angiotesin I. Therefore, it is proposed that ET-1 may play a role by autocrine or paracrine in the human normal adrenal gland. PMID- 9208580 TI - [The imaging features of multiple endocrine neoplasia]. AB - The article describes ultrasonographic (US), computed tomographic (CT), 99mTc MIBI and angiographic findings of 16 cases of multiple endocrine neoplasia (MEN) confirmed by operations and pathological examinations. Hyperparathyroidism either was the first manifestation of MEN I or was diagnosed simultaneously with other tumors. Medullary thyroid carcinoma usually developed first or coincided with pheochromocytoma in MEN I. Regular US and CT screening can lead to the detection of new endocrine neoplasia. The authors consider that US and CT are the best imaging methods for MEN. PMID- 9208581 TI - [Experimental study and clinical application of primary choledochorraphy after choledochotomy]. AB - An animal model was established in dogs to compare primary choledochorraphy with the traditional T-tube method. Authors observed cholangic histologic changes under microscope and electron-microscope between the two groups. The result showed that primary choledochorraphy was better than the traditional method. The advantage of primary choledochorraphy lies in its simplicity, safety and reliability, which have been shown in 30 patients of cholelithiasis in clinical application. It is suggested that for patients primary choledochorraphy may clesirably be used for patients who are indicated for this approach. PMID- 9208582 TI - [The pathology of labial salivary gland in the diagnosis of Sjogren's syndrome]. AB - To explore the sensitivity and specificity of the pathology of labial salivary gland in the diagnosis of Sjogren's syndrome (SS) and its specific pathological changes in SS, 100 labial salivary gland specimens were observed and assayed in our hospital. The results indicated that there was no significant difference in acinar atrophy, duct dilatation and lymphocytes infiltration between primary SS (1 degree SS) and secondary SS (2 degrees SS) patients. But those changes had significant difference between 1 degree SS and non-CTD patients (P < 0.01). It was also found in our study that Chisholm grade IV was achieved in 78.57% 1 degree SS specimens, 65.22% 2 degrees SS specimeus and 22.22% in non-CTD specimens, respectively. According to Chisholm standard, the sensitivity and specificity of pathology in the diagnosis of these SS patients was 78.8% and 77.1%. Our patients were still classified by using Chisholm standard. If focal lymphocytes infiltration was used as the single criterion to SS diagnosis, its specificity was not high. In the diagnosis of SS authors should consider the pathologic changes of salivary gland as well as the clinical information in order to avoid misdiagnosis. PMID- 9208583 TI - [The role of Ca2+ in the pathogenesis of human pituitary GH-secreting adenomas]. AB - Effects of Ca2+ channel blockers (nicardipin and nifedipin) and Ca2+ ionophore A23187 on the basal secretion and on the secretion stimulated by GRH or inhibited by SMS, a SRIF analogne of GH were investigated in monolayer cell cultures of 23 cases of human pituitary GH-secreting adenomas. The roles of GRH and SMS in 45Ca influx were investigated also. The GH secretion of most GH adenomas was depended on Ca2+, but the abnormality in different link of GH secretion mediated by Ca2+ was observed. The defects of receptor and post-receptor including Ca2+ channel and Ca(2+)-GH secretion couple regulated by GRH and SRIF were found in 66.7% and 55.6% of GH adenomas respectively. These abnormalities may contribute to GH hypersecretion in GH adenomas. PMID- 9208584 TI - [The hepatitis A virus isolated and adapted in human diploid fibroblast cells (KMB17)]. AB - Three hepatitis A virus (HAV) strains (L8, H32 and H52) were isolated from stool specimens of hepatitis A patients during an epidemic in 1988 in Shanghai and were adapted to grow in human diploid fibroblast cell strain (KMB17). The strains were serially carried through 10 passages in KMB17. The infectious titrations of these virus strains cultured for 28 days at the 10th passage were high up to 7.0-7.75 log TCID50/ml. The maximum virus yields occured in the 28th day after infection. Immune electronic microscopic studies revealed that the viral particles were 27 30 nm in diameter. The strains were neutralized by specific anti-HAV serum. Special fluorescent stained particles were observed clearly in the infected cellular plasma by direct IF. The above data suggest that the three strains are HAV and may be useful for further study. PMID- 9208586 TI - [Changes in calcium uptake and Ca(2+)-ATPase activity of cardiac sarcoplasmic reticulum in rat associated with hypoxia of various degrees]. AB - Two groups of Wistar rats were exposed to hypoxia at 5000 to 8000 m high above the sea level in a hypobaric chamber for 7 days. The effects of hypoxia on left cardiac function, sarcoplasmic reticulum (SR) Ca2+ ATPase activity and SR calcium uptake were observed. The results indicated that the left ventricular function exposed to hypoxia at 5000 m was much higher than that at 8000 m. Calcium uptake and Ca2+ ATPase activity of the exposed groups decreased significantly compared with those of the control group, but they were much higher in the exposed group at 5000 m than at 8000 m. These results suggest that the changes in calcium uptake and Ca2+ ATPase activity of SR may be one of the important biochemical indicators for cardiac function alterations associated with hypoxia. PMID- 9208585 TI - [Cordocentesis for prenatal diagnosis of fetal diseases]. AB - For the purpose of prenatal diagnosis, 30 fetal blood samplings were performed in 30 pregnant women from 19 to 36 gestational weeks. The samples were taken with a 22-gauge needle guided by ultrasound. Twenty-five cordocenteses were succeeded at the first attempt. A second attempt was needed in other 5 cases. The duration of procedure was less than 15 minutes in 83.3% cases. 1-6 ml fetal blood was obtained for each case. No lethal complication occured in our series. It is believed that cordocentesis could be a useful tool for prenatal diagnosis, and the intrahepatic-vein sampling is a preferable alternative choice if cordocentesis fails. PMID- 9208587 TI - [Effect of the T4 on the viscera and embryo of perinatal mice]. AB - 58 Female Kun Ming mice of perinatal stage (from the 15th day of pregnacy to the 21th day after birth) were fed with Tripcholorolide (T4) isolated from multiglycosides of Tripterygium wilfordii (GTW) at a doze of 0.6 mg/kg for group 1 or 0.3 mg/kg for group 2 per day for 4 weeks. Lactation was decreased in some females and some F1 off spring died. The succinic dehydrogenase (SDH) activity of the mice liver were increased due to the destruction of its mitocondian. Liver cells degenerated and glycogen decreased. Distal tubules of kidney degenerated. Heart and spleen were normal. T4 was also fed to 10 female mice from the 5th to 17th day of pregnacy. However, neither the absorbed fetus nor dead fetus increased. PMID- 9208588 TI - [Subcellular localization of blood group substances ABH in human gastrointestinal tracts]. AB - Authers investigated subcellular localization of ABH substances in normal human stomach, duodenum and transverse colon by post-embedding immunogold method using monoclonal antibodies specific for A, B and H antigens. The immunoreactivity was usually restricted to the following organelles: mucigen granules of epithelical cells of gastric mucosa, zymogenic granules of chief cells of the gastric gland, both mucigen granules and Golgi cisternae of neck mucous cells of the gastric gland and of epithelial cells of the duodenum mucosa as well as the Brunner's gland. ABH antigens were also visualized in microvilli of absorptive cells and mucigan granules of goblet cells of the transverse colon. The results suggested that ABH substances in those cells were produced in Golgi cisternae and were transfered to secreting granules, then were secreted with mucus. The mosaic distribution of ABH substances in cells and the effect of embedding resins on ABH antigens and the ultrastructure of cells were discussed. PMID- 9208589 TI - [Studies on biological characteristics of HL-60 and all-trans-retinoic acid resistant HL-60 cells--comparison of the basic biological and cytogenic features of the two cell lines of HL-60]. AB - The biological features of HL-60 and all-trans-retinoic acid resistant HL-60 cells (HL-60/ RA) were studied in terms of cell proliferation, cytokinetics, chromosome distribution and G-band karyotypes. Results indicated that the two cell lines exhibited similar morphology and doubling time and similar cytokinetic features, but the cell population in S phase of HL-60/RA is smaller than that of HL-60 cells. Their chromosome number is around 45 +/- 3 for HL-60 and 45 +/- 1 for HL-60/RA cells. G-band karyotype analysis showed that they have a similar diploid karyotype and some abnormal chromosomes. These results demonstrated that HL-60 and HL-60/RA cell lines were originated from the same source. PMID- 9208590 TI - [The study of BL21(DE3)/PET-11 as an expression system of human IL-2]. AB - The coding region of hIL-2 cDNA is specifically amplified by PCR. A Nde I site is introduced into its 5'-end and its 3'-non coding region is partly deleted. After filling-in by Klenow enzyme, the DNA fragment is cloned into Sma I site of PUC18. Eight white colonies are screened and one (C1) contains Nde I site. DNA sequencing shows that non-specific mutation has not been found in its coding region. The DNA fragment digested by BamH I, Nde I of C1 plasmid is recovered and subcloned into PET-11 and transfered into BL21 (DE3). After IPTG induction, the Bacteria lysate is run directly on SDS-PAGE. Western blot test shows a specific rhIL-2 McAb binding band. The molecular weight is about 17000. Laser scanning indicates that the absorption area of IL-2 is about 5% of the total area. PMID- 9208591 TI - [CuZn-SOD determination of sera in patients with rheumatic diseases]. AB - Superoxide anion (O2.-) plays an important part in reactive oxygen species (ROS). In order to explore its effect on the pathogenesis of rheumatic diseases, authors had determined CuZn-SOD contents of sera in 132 subjects involving the patients of rheumatic diseases (SLE, RA, etc), non-rheumatic diseases and normal controls by using enzyme linked immunosorbent assay (ELISA). The results showed the followings: CuZn-SOD contents of 27 normal subjects: 98.80 +/- 20.74 ng/ml (x +/- s); that of 27 non-rheumatic diseases cases: 72.24 +/- 16.60 ng/ml (x +/- s); of 22 SLE cases: 56.56 +/- 19.27 ng/ml (x +/- s); of 27 RA cases: 61.56 +/- 20.53 ng/ml (x +/- s); of 29 other rheumatic diseases cases: 68.97 +/- 17.79 ng/ml (x +/- s). Statistical test was made: both CuZn-SOD contents of rheumatic disease and non-rheumatic disease were lower than that of normal subjects with more significant difference (P < 0.001); compared with that of non-rheumatic diseases patients, SLE cases had significant difference (P < 0.01); RA cases had significant difference (P < 0.05); other cases of rheumatic diseases had no statistical differrence (P > 0.05). Above results suggest that superoxide anion is a non-specific inflammatory mediator which contributes to disorders with inflammatory damages (rheumatic or non-rheumatic diseases), where CuZn-SOD content tested was obviously lower than normal subjects; among the rheumatic disease patients, CuZn-SOD contents of the sera of SLE patients were the lowest because of its more autoimmune antibody, more severe inflammatory and immunological reaction. This work laid the theoretical and experimental foundation for the clinical application of exogenous CuZn-SOD in the treatment of rheumatic diseases. Combined use of CuZn-SOD scavengers may get better result because of the complexibility of ROS inflammatory mechanism. PMID- 9208592 TI - [Spinal cord monitoring during spinal surgery]. PMID- 9208593 TI - [Effect of phosphorylation by protein kinase C on the DNA binding activity of high mobility group protein I]. AB - High mobility group protein I (HMG-I) is a nonhistone chromosomal protein. The present study aims to examine phosphorylation of HMG-I by protein kinase C (PKC) and its effect on HMG-I's DNA binding activity. HMG-I, extracted and purified from rat brain was phosphorylated in vitro equally well by PKC alpha, beta, gamma and delta. Phosphoamino acid analysis indicated that both serine and threonine residues were phosphorylated. The nonphosphorylated HMG-I was shown to bind specifically to the fragment of DNA containing bp -708 to -458 of RC3 genomic DNA, which is abundant in A-T sequences. In contrast, phosphorylation of HMG-I by PKC resulted in an attenuation of binding to the DNA fragment. It is suggested that phosphorylation of HMG-I by PKC may regulate DNA binding activity of HMG-I, thereby possibly altering its biological functions. PMID- 9208594 TI - [Induced-expression of MHC-DR molecules on mice islet cells]. AB - The correlation of induced-expression of MHC-DR molecules with insulin release in cultured mice islet cells has been studied. The results showed that, (1) No detectable induced-expression of MHC-DR molecules by TNF-alpha alone at the concentration of 50-100 U/ml has been assayed, but the induced-expression of MHC DR molecules has been identified in a few islet cells (8.5 +/- 2.9%) with IFN-r alone at concentration of 50-100 U/ml. (2) The induced-expression of MHC-DR molecules has been detected in as high as 80% of cultured islet cells with TNF alpha and IFN-r combined at different concentrations. (3) Insulin released from cultured islet cells was enhanced by IFN-r and TNF-alpha either alone or combined at different concentrations. These results suggest that the induced-expression of MHC-DR molecules themselves does not damage the function for insulin release in cultured islet cells. PMID- 9208596 TI - [Detection of c-Ki-ras oncogene codon 12 point mutations in lung carcinoma, gastric carcinoma and hepatic carcinoma]. AB - DNA extracted from paraffin-embedded tissues of 30 lung carcinomas, 40 gastric carcinomas and 22 hepatocellular carcinomas was tested for the presence of c-Ki ras codon 12 point mutation by PCR-RFLP technique. The results showed that 20% (6/30) lung carcinomas and 2.5% (1/40) gastric carcinomas had Ki-ras codon 12 point mutation, but non of the 22 hepatocellular carcinomas displayed this point mutation. Among 6 cases of lung carcinomas of mutation, 4 were squamous carcinomas and 2 were adenocarcinomas. PMID- 9208595 TI - [Effect of dexamethasone on the human interleukin-2 receptor alpha chain gene expression]. AB - Glucocorticoids have wide effects on the immune system, the mechanisms of these effects have not been clearly defined. The interaction of interleukin-2 (IL-2) with its receptor (IL-2R) is a critical control point in the T-cell mediated immune response. The effect of dexamethasone, a synthetic glucocorticoid hormone, on IL-2R is unclear in comparison with its inhibition on the accumulation of IL-2 mRNA. Our study shows that dexamethasone increases the basal and PHA-induced mRNA levels of IL-2R alpha gene in Jurkat cells, but it does not affect the expression of reporter gene constructs containing 5' flanking sequences (-482bp/ +16bp or 350bp/+16bp) of the IL-2R alpha gene in PHA activated Jurkat cells. These results indicate that dexamethasone plays a positive regulatory role in the IL-2R alpha gene expression, but the effect is not mediated by the previously identified proximal regulatory elements located between -482bp/+1bp. Meanwhile, our results also provide evidence that the activation mechanism of dexamethasone is different from that of PHA, and that dexamethasone and PHA show a synergistic effect in the induction of IL-2R alpha gene expression. PMID- 9208597 TI - [The postnatal changes of follicle-stimulating hormone receptor mRNA level in rat testis]. AB - It was found in this study that the follicle-stimulating hormone receptor (FSHR) mRNA levels varied between days 2 and 60. The FSHR mRNA rised slightly in the neonatal rat testis and then declined. In the prepuberty the content of FSHR mRNA remained at a Constant level. During the puberty the FSHR mRNA increased to its highest level on days 25 and finally it rised again after days 40. PMID- 9208598 TI - [Alteration of mRNA transcripts of rat Sertoli cell secreted proteins and Leydig cell LH/CG-R protein following glycosides of Tripterigium Wilfordii Hook administration]. AB - Our studies were undertaken to assess the male antifertility mechanism of the total glycosides extracted from Tripterigium Wilfordii Hook. f. (GTW). In experimental group I, GTW was given by gastric gavage at a dose of 30 mg/kg per day for 35 days. In experimental group I, GTW was given in the same way at a dose of 10 mg/kg per day for 49 days. The transcripts level analysis indicated that in all the experimental groups ABP, SGP-2 and LH/CG-R mRNA were increased. Two ABP and two LH/CG-R mRNA transcripts with respective molecular sizes of 1.7 kb, 2.3 kb and 1.2 kb, 4.4 kb were identified. ABP and LH/ CG-R mRNA with 1.7 kb and 1.2 kb increased, while those with 2.3 and 4.4 kb remained unchanged. In addition, the changes of different proteins at the mRNA level were shown to be GTW dose dependent. The morphological features of germ cell degeneration and appearance of germ cells in tubular lumen suggest that the increases in transcripts level of three proteins in testis may not be directly associated with the effect of GTW. The question whether GTW interfere with the function of sertoli cells and leydig cells would be study. PMID- 9208599 TI - [Effects of curcumin derivatives on the GJIC of normal and tumor cells]. AB - Experiments were conducted by using scrape-loading and dye transfer (SLDT) method to study the gap junction intercellular communication (GJIC) of Chinese hamster lung cells (V79), mouse fibrous cells (Balb/c-3T3), rat liver cells (WB) and human embryonal lung cells (2BS). We also observed the inhibition of the GJIC by TPA and the antagonistic effect of Curcumin derivatives on TPA. The results indicated that V79, WB, 3T3 and 2BS normal cells showed medium level of GJIC, and TPA could inhibit the GJIC to some extents. Curcumin derivatives (91022, 91022-S) could counteract the inhibition of TPA-induced GJIC. It was also found that human lung adenocarcinoma cell (A549) and GLC lacked GJIC, and 91022 could improve the GJIC of A549 cell. It may be related to its anticancer activity. PMID- 9208600 TI - [Effect of IL-4 on biological regulation of tumor cell lines]. AB - The effect of IL-4 on biological regulation of tumor cell lines was observed by using human recombinant interleukin-4. The observation included cell proliferation, differentiation, cell surface marker and cytokine secretion. The effect of IL-4 on cell proliferation of nine cell lines was different. It showed 4/9 inhibitory effect, 3/9 enhancing effect and 2/9 no effect. The inhibitory effect of IL-4 on U937 cell proliferation was correlated with cell differentiation. IL-4 enhanced the expression of cell surface markers (including MHC class I, MHC class II and TAA) of some cell lines. Furthermore, IL-4 inhibited the production of IL-2 by MLA144. The preliminary results indicate that the effect of IL-4 on biological regulation of tumor cell lines is benefitial for controlling tumor cell growth and enhancing tumor cell recognition by effector cells. However, the down regulation also could be exerted. PMID- 9208602 TI - [The effect of ligustrazine and 764-3 on type I and type III collagen mRNA expression in fibroblasts]. AB - The effect of ligustrazine and 764-3 on type I and type II collagen mRNA expression in cultured fibroblasts was studied. The results showed that ligustrazine at a final concentration of 30 micrograms/ml for 24 hours could inhibit type I and type II collagen mRNA expression, 764-3 at a final concentration of 15 micrograms/ml for 24 hours could stimulate type I collagen mRNA expression, but exerted no effect on type II collagen expression. The possible mechanism of 764-3 action was discussed. PMID- 9208601 TI - [The pharmacokinetics of pancuronium bromide in infants, children and adults]. AB - Twenty-four patients scheduled for elective plastic surgery were selected to study the pharmacokinetics of pancuronium bromide under enflurane anesthesia. The patients were divided into three groups by their ages; Group 1 consisted of 5 infants (0.75-2.95 years); Group 2 contained 13 children (4-14 years); Group 3 included 6 adults (16-27 years). An improved fluorimetric assay was used to measure the plasma concentrations of pancuronium bromide after administration of pancuronium bromide at a dose of 100 micrograms/kg. The results showed that the disposition of pancuronium bromide may be best described mathematically by a two compartment open model in all patients. The younger the patients, the larger the distribution volumes and the higher the Cl. There were significant differences among the three groups with regard to V1, V2, Vdss, Cl, AUC. The T1/2 beta and MRT were longer in Group 1 than in Group 2 and Group 3. PMID- 9208603 TI - [Preliminary investigation of Helicobacter pylori infection in Linqu County of Shandong province]. AB - A preliminary epidemiological investigation of Helicobacter pylori (HP) infection was performed with 13C-urea breathing test for 218 residents at the age from 40 to 69 years in a high risk area of gatric cancer, Linqu County of Shandong province. Our results show an overall HP infection rate of 71.10%, or 69.49% for men and 73.00% for women. The HP infection rate in the group of serious chronic atrophic gastritis was significantly higher than that of mild chronic atrophic gatritis (P < 0.05). The HP positive group also has a higher rate of complaining about gastric discomfort than the HP negative group (P < 0.02). PMID- 9208604 TI - [Effect of tetramethyl pyrazine on coronary vasoconstriction induced by endothelin-1 in dogs]. AB - The purpose of this study was to determine the antagonistic effect of tetramethyl pyrazine (TMP), a sort of chinese herbal medicine, on coronary vasoconstriction induced by endothelin-1 (ET-1) in closed chest dogs. ET-1 at doses of 50, 75 and 100 pmol was selectively administered into left main coronary artery and coronary angiogram was performed in 1, 3 and 10 minutes after intracoronary administration of ET-1. After a 60 minute interval ET-1 administration and coronary angiogram were repeated in two groups in group A with 5 dogs intravenous infusion of saline solution was administered while in group B with 4 dogs TMP was infused at a dose of 80 mg/kg. Blood pressure of intra-femoral artery, heart rate and ECG were monitored during the experiment. The study demonstrated that coronary vessel diameter significantly decreased by 17% (P < 0.02) in group A and 20% (P < 0.02) in group B, associated with ischemia in ECG (4/5 in group A and 3/4 in group B) after intracoronary administration of ET-1. Endothelin-1 induced coronary vasoconstriction and ischemic changes in ECG were significantly inhibited by intravenous TMP. The coronary diameter increased by 20% (P < 0.03) after administration of TMP, comparing with the control group. Heart rate had an increased response to TMP. In conclusion this study demonstrated that intracoronary administration of ET-1 caused significant myocardial ischemia through coronary vasoconstriction, which was inhibited by TMP. TMP significantly dilated coronary artery. PMID- 9208605 TI - [The effects of a single active ingredient (T4) of Tripterygium Wilfordii Hook on the production of tumor necrosis factor by the peripheral blood mononuclear cells and synovium cells of rheumatoid arthritis patients]. AB - Encouraged by good therapeutic results of the semipurified preparations of Tripterygium Wilfordii Hook (TWH) (code name T4) a single-active ingredient of TWH with the code name of T4 was obtained. The in vitro effects of T4 on the bioactivity of TNF produced by PBMC of normal subjects (n = 10) and RA patients (n = 6) or the RA digested synovium single cells (DSSC) were studied. The results showed that under pretreatment of PBMC of healthy persons with T4 (35 ng/ml) for two hours, TNF were reduced to 36.13 +/- 1.75% in comparison with the control of 63.33 +/- 2.51% (P < 0.001), with an inhibition rate of 40.31%. Similarly, the inhibition rate for the PBMC of RA patients (P < 0.001), was 25.54%. The fact that coculture of PBMC and T4 for 6 hours instead of treatment with T4 for 2 hours caused an inhibition rate of 40.51% (P < 0.01), might indicate that time of interaction was also a factor of importance. T4 also reduced TNF secretion by RA DSSC from 72.1 +/- 6.1% to 51.6 +/- 2.0% with an inhibition rate of 28.4%. The aformentioned indicates that T4 appears to possess suppressive actions on production of TNF by PBMC and DSSC of RA patients. Therefore, it is possible that T4 may be used clinically for RA patients. PMID- 9208606 TI - [Phase I clinical study of a new anticancer drug boanmycin]. AB - From August 1993 to March 1994, 36 patients were enrolled for a phase I study of Boanmycin (Bleomycin A6) a new anticancer drug to determine it toxicity and maximal dose. Of the 36 cases, 16 were male and 20 female, age 20-62. Dose escalation was performed if a minimum of threed patients were fully evaluable for toxicity (2 weeks following drug administration) and if severe or life threatening toxicity had not occurred. Dose of Boanmycin were escalated from 1 mg (0.5-0.7 mg/m2) to 12 mg(6.7-7.5 mg/m2) i. m. three times every week for two weeks. Surveillance of serum concentration of Boanmycin was conducted in six cases by microbiological analysis, and the pharmacokinetics parameters were obtained. Phase I study of Boanmycin showed that Boanmycin had no myelosuppression and cardiac toxicity, and its major adverse reactions were fever, gastrointestinal reactions and hardening at injection (by i. m. route). Rather than dose-related, fever was individual-related. There were mild myalgia, alopecia, skin rash, pigmentation in some patients. All adverse reactions resolved after discontinuation of therapy. There was no Boanmycin-related lethal complication, and the maximum tolerated dose was not obtainable. If patients have renal or lung disease, Boanmycin aggravate their renal and lung functions. Therefore we recommend that dose of Boanmycin for phase I clinical trial should be is 8-10 mg(5-6 mg/m2) i m or iv (iv can decrease local side effect) two-three times per week. PMID- 9208607 TI - [Image morphological quantitative analysis of pathological changes of intra acinar arteries in experimental pulmonary artery hypertension and outcomes of treatment]. AB - The relative ratios of vessel lumen area/total area (EA/TA), vessel wall area/total area (MA/TA) and vessel wall thickness/vessel external diameter (WD/TD) of intra-acinar arteries were studied with image analysor in 35 rats of experimental pulmonary artery hypertension and outcomes of treatment. All the rats were divided into control group (R0), hypertension group (R1) and treated groups (R2), (R3) and (R4). The results showed relative ratios of EA/TA decreased significantly in the palmonary artery hypertension group in comparison with the control and treated groups, while the values of WA/TA and WD/TD increased significantly. The results may be valuable for morphological quantitative study, diagnosis of pulmonary artery hypertension and prediction of the disease prognosis. PMID- 9208608 TI - [The pulmonary function in scoliotic patient]. AB - This paper investigated preoperative pulmonary functions in 134 scoliotic patients, of whom 32 cases was followed up for their postoperative pulmonary functions. The results show that the bigger the Cobb of the scoliosis, the greater the damages of the pulmonary functions. Congenital scoliosis is more severely affected by pulmonary functions than idiopathic scoliosis. There was no significant difference among the different sections of scoliosis with regard to their effect on the pulmonary functions. The authors concluded that the Harrington Instrumentation improved pulmonary functions in scoliosis. PMID- 9208609 TI - [Selective training of the vastus medialis muscle using electrical stimulator for chondromalacia patella]. AB - Chondromalacia patella is closely related with subluxation and tilt of patella, as well as with muscular atrophy of quadriceps, especially in vastus medialis muscle. 364 cases of chondromalacia patella were treated with selective training of the vastus medialis muscle using electrical stimulator in our hospital. 211 cases were followed up after treatment from 6 months to 3 years. Among them excellent and good results were seen in 130 cases (62%), fair results were seen in 69 cases (33%) and no change was seen in 12 cases (5%). Significant reduction of CA (P < 0.01) and LPA (P < 0.001) were observed in all these patients in comparison with their primary angle. We believe that the selective training of the vastus medialis muscle using electrical stimulator is one of the effective methods for the treatment of chondromalacia patella. PMID- 9208610 TI - [Effects of temperature on the viability and infectivity of preparasitic larvae of Romanomermis yuanenesis]. AB - Romanomermis yuanenesis (Mermithidae: Nematoda) was found in Henan, China (Song and Peng, 1987), which has a broad host range in Culicinae mosquito and has been used successfully in field test for control of culex tritaeniorhynchus, culex fatigans and Aedes albopictus in Sichuan, Yunnan, Guangxi and Henan Provinces. This study was attempted to determine the viability and infectivity of preparasitic larvae in various temperatures. The cultures containing R. yuanenesis eggs were flooded 2h with distilled water, filtered and blocked with 1% agarose. Put the filter paper into water, then the motile preparasites separated from the unhatched eggs and got through the agarose membrane into water. About 200ml water containing preparasites free from eggs were held at 26 degrees C-28 degrees C, 16 degrees C-18 degrees C and -2 degrees C to 2 degrees C for test. The motility or lack of motility was used as the criterion to distinguish the living and dead nematodes. The rate of infection of mosquitoes and the rate of parasitism of nematodes were used to show the infectivity of the preserved preparasites. The results showed that at -2 degrees C to 2 degrees C, more than 90% of preparasitic larvae of R. yuanenesis survived for 8 days and the rate of mosquito infection was 87.5% to 100%, but at 26 degrees C-28 degrees C and 16 degrees C-18 degrees C the survival times of 90% preparasites were only 24 hours and 48 hours respectively. It indicates the low temperature preservation may prolong the survival time and keep the infectivity of these preparasitic larvae. PMID- 9208611 TI - [Carcinogenic mechanisms of multiple genes in cervical carcinoma]. AB - The alterations of multiple genes and their carcinogenic mechanism in cervical carcinoma were studies by molecular hybridisation, PCR and PCR-ASO techniques. The G-T point mutation in the 12th coden of Ha-ras was detected in cervical carcinomas with mutation frequency of 18.2% (8/44), and the amplification rate of Ha-ras gene was 45% (9/20). The c-erb B2 was amplified 3-30 fold with an amplification rate of 73.3% (11/15) in cervical carcinomas and 5 cancerous samples showed gene rearrangement. The elevated copies of c-myc gene with amplification rate of 91.7% (11/12) were observed in cervical carcinomas. The study of HPV16 viral gene showed that the existence of HPV16 DNA sequence was positively associated with c-myc gene amplification in cervical cancerous samples. The p53 and Rb tumor suppressor genes absence of deletion were observed in the 12 specimens of cervical carcinoma investigated. As mentioned above, the study on alteration and carcinogenic mechanism of multiple genes indicated that 3 oncogenes and HPV16 viral gene were activated or integrated throygh different mechanisms and they played roles in co-carcinogenesis. The integration of HPV16 gene might promote the c-myc gene at the early stage in carcinogenesis of cervical carcinoma, while the alteration of Ha-ras and c-erb B2 gene might be middle-late event. As for the roles of the p53 and Rb tumor suppresor gene in cervical carcinogenesis need further researches. PMID- 9208612 TI - [The PCR amplification, cloning, sequencing, expression in E. coli of gene encoding endoflagella subunit protein (fla B) from Leptospira interrogans serovar lai]. AB - A pair of oligonucleotide primers were designed by ourselves to amplify the endoflagella gene of L. interrogans serovar lai. A fragment about 840 bp was generated with PCR and inserted into plasmid pUC8 after the fragment and pUC8 were digested respectively with Bam HI and Pst I. A recombinant plasmid (designated as pLF1) was obtained. SDS-PAGE analysis indicated that a 33 kd was expressed in E. coli JM103 harboring pLF1 and the expression level of the protein was 11% of total bacterial soluble proteins. Western blot analysis showed that the protein band could be recognized by the antiserum against the endoflagella (Axiall filament) of Leptospira interrogans serovar lai. Nucleotide seguence data showed an open reading frame encoding 282 aminoacids residues, corresponding to a protein of molecular weight 33.6 kd. The G + C content of endoflagella subunit protein gene was 48 mol%. Therefore, the G + C content of the leptospiral fla B Gene is significantly higher than the reported 39 mol% G + C content of leptospiral genome of L.interrogans serovar lai but similar to the G + C of the Treponema pallidum genome. Comparison of the deduced endoflagellar subunit protein (fla B) amino acid sequence with flagellins from other bacteria revealed a high level of identity with the Treponema pallidum fla B proteins. Immunization/protection experiment was performed on the model of BALB/c mice and showed that the survival rate in the group JM103-pLF1 was higher than that in the group JM103-pUC8, but statistically the difference between them was significant (P < 0.05) and pLF1 did not induce significant levels of agglutinating antibodies against L.interrogans serovar lai. PMID- 9208613 TI - [16S rRNA gene PCR-SSCP analysis of the reference strains from 15 serovars (14 serogroups) of pathogenic leptospires in China]. AB - The DNAs of reference strains from 15 serovars (14 serogroups) of pathogenic leptospires in China were amplified with 16S rRNA gene primers, and then single strand conformation polymorphism (SSCP) of the products were analyzed in 12.5% nondenaturing mini polyacrylamide gel (containing 5% glycerol) combining with silver-staining. All products showed two bands on electrophoresis at different parameters of voltage or current and concentration of gel. It proved that serovar lai, serovar canicola, serovar pyrogenes, serovar autumnalis, serovar australis, serovar pomona, serovar linhai, serovar hebdomadis, serovar haemolytica, serovar wolffi and serovar paidjan have the identical pattern (Leptospira interrogans), while serovar javanica, serovar ballum, serovar tarassovi and serovar manhao I belong to another pattern (L. borgpetersenii). The result was consistent with the classification of genetic species by Yasuda et al (1987) and Ramadass et al (1992). PMID- 9208614 TI - [Simultaneous phenotyping of proteins with various pHs by complex IEF and its application in forensic haemogenetics]. AB - We have developed a new technique of complex isoelectric focusing (IEF) by which various proteins with different pHs can be detected out in polyacrylamide gels containing ampholytes with different pH ranges at the same time. The success of phenotyping 12 human proteins by this technique proved it was reliable and efficient. The cumulative Epp and the cumulative Dp reached 0.9 and 0.9999 respectively. It provides a new approach to analysis of genetic markers in the field of forensic haemogenetics. PMID- 9208615 TI - [Comparison of diet, physical activity and serum lipids between Chinese and Japanese: Chengdu-Fukuoka Study]. AB - A detailed diet history was taken from 326 Chinese adults, 116 college students in Chengdu, China, and 300 Japanese adults and 4529 college students in Fukuoka, Japan. Both populations consume high carbohydrate and low fat diet, but the Chinese diet was higher in carbohydrate, cereal energy/total energy ratio and polyunsaturated fatty acids/saturated fatty acids ratio, and lower in protein, fat and animal fat/total fat ratio compared to the Japanese diet. Cholesterol intake was significantly lower in Chinese than in Japanese (P < 0.001). Physical activity index in Chinese was higher than that in Japanese. Serum lipids and lipoprotein cholesterols were measured in the adults of Chinese and Japanese. Except the serum triglycerids level in female, the total cholesterol, TG, high density lipoprotein-cholesterol and low density lipoprotein-cholesterol levels in Chinese adults were lower than those in Japanese adults (P < 0.001). Since cholesterol is the major predisposing factor for coronary heart disease and the results of our study show that a high carbohydrate and low fat diet can be effective in lowering the cholesterol level in blood it will be of interest to see which kind of oriental dietary may be a preventive against coronary heart disease. PMID- 9208616 TI - [Relationship between SCE points and G-banding location in patients with nasopharyngeal carcinoma]. AB - Using a technique for simultaneous demonstration of G bands and SCE, we analyzed the SCE frequencies, cell cycle kinetics, distribution of SCE points and "hot spots" which correlate with fragile sites, proto-oncogenes locus in cultured peripheral blood lymphocytes in 20 untreated patients with nasopharyngeal carcinoma (NPC). The SCE frequencies were found to be higher in patients as compared with controls (P < 0.01). The SCE points of patients were significantly higher than those of controls in chromosome 1,2,3,4,5,7,8,12,13,14,17 (P < 0.05), indicating the chromosome instability of NPC patients. We identified 17 points as high frequency SCE hot spots, and found that 29.41% SCE hot spots fragile sites and proto-oncogenes were localized at the same bands. The distribution of SCE points and chromosome structure were discussed. PMID- 9208617 TI - [Establishment of an effective way for separating and culturing tumor infiltrating lymphocytes]. AB - The key to preparing TILs for clinical application is to enhance its capacity of proliferation and cytotoxicity. For this purpose we have made a study of methodology and established an effective way consisting of 3 processes to separate and culture TILs. It included: (1) mechanical splitting; (2) digesting with 0.05% collagenase I and 0.003% DNAase I mixture in room temperature for 6 hrs and 18hrs in cool, and (3) centrifuging with 75% and 100% Ficoll non continuous grade density. The separated TILs were suspended in conditional culture medium for proliferation. Most of them reached the amount of 10(9) at the end of culture, which was essential for clinical therapy. Their NK and LAK activities were 56.3 +/- 11.7% and 48.6% +/- 10.6% respectively, in which the CD6+ T cells played an important role. The results suggest this a perfect way for separating and culturing TILs. PMID- 9208618 TI - [Monoclonal antibodies to human progesterone receptors: production, indentification and clinical application]. AB - A peptide with 15 amino acid of human progesterone receptor (PR) was synthesized and used as antigen. Five hybridoma cells were obtained by cloning technique. In tests to hybridoma cell H4, the site specificity of the antibody to PR was found. The antibody reacted specifically with PR, but did not cross-react with ER, AR, or GR. Five women who used clomiphene citrate (CC) 100mg/d x 5 days were involved in a study of serum pregesterone (P) and endometrial histology in early luteal phase. There was no difference in serum P level in two groups. Three cases in CC group showed abnormal secretory endometrium. PR concentration in the nuclei of glandular cells of endometrium in CC group was significantly lower than that in the control group. In women with abnormal secretory endometrium, PR concentration in the nuclei of glandular cells was lower than that in women with normal endometrial histology. CC can influence endometrial histology by changing-PR concentration in the nuclei of glandular cells. The endometrium can not respond to the stimulus of P at normal levels and thus results in luteal phase defect. PMID- 9208619 TI - [Immunohistochemical detection of antigen of human cytomegalovirus in hepatocyte]. AB - The expression plasmid of human cytomegalovirus (HCMV) 52kd glycoprotein was induced and amplified, and the recombinant protein was extracted. The rabbit antiserum to the recombinant protein was prepared. The antiserum-reacted to the antigen of HCMV AD165, and the titer of antiserum was 1 : 3200 by using 125I Protein A. Using Avidin-Biotinperoxidase complex technique (ABC method), we detected the HCMV 52 kd glycoprotein in the hapatic slides of 9 cases. Among them, one case (an infant died of congenital heart disease) was held as a negative control. The other 8 cases (patients with clinically diagnosed cytomegalic inclusion disease) had viral hepatitis histopathologically. Of these 8 cases, 7 had positive signals but found in the nucleus and cytomembrane of the hepatocytes, no positive signal was found in the hepatocytes of the control. The result showed that HCMV 52kd glycoprotein was located in the nucleus and on the membrane of the infected hepatocyte. The antibody prepared with the antigen of gene engineering could find the virus antigen and represent the virus gene state which was active or silent. ABC method is not only simple, sensitive and specific, but also effective in localization. It is of importance to clinical diagnosis, differential diagnosis and prognostication. PMID- 9208620 TI - [The influence of HX- I on rabbit thyroid allografts]. AB - We studied the anti-rejection effect of HX- I, a preparation of traditional Chinese herbs, on rabbit thyroid allografts. The transplantations were performed on 28 rabbits after total thyroidectomies. The grafting sites were in their pretrachial muscles. These animals were divided into four groups, namely, Group I: homografts: Group I: allografts without medication; Group II: allografts with dexamethason (0.25 mg/(kg.d) intramuscularly), and Group IV: allografts with HX-I water solution, (5g/(kg.d), peros). The medication lasted 28 days. Blood samples were drawn every week postoperatively. Serum T3 and T4 were tested by RIA. The grafts were removed for histopathological evaluation on the 28th day postoperatively. The histopathology of rejection and survival were scored and classified. On the 7th and 14th days, serum T3 and T4 levels were almost the same between groups. On the 21st and 28th days, the T3 and T4 levels were higher in Groups I and IV than those in Group II (P < 0.05). The histopathological findings were; in Group I, damaged follicles with much lymphocytes infiltration and fibrosis, and 6 cases being rejected; in Group II, two deaths and three cases with damaged thyroid tissue and much lymphocytes infiltration; in Group IV, three cases with damaged thyroid tissue and four intact grafts. Our results indicate that HX-I and dexamethason both can inhibit rejection in thyroid allografts in rabbits, but dexamethason has more side effects HX-I has many components and the machanism of its early anti-rejection effect is worthy of further study. PMID- 9208622 TI - [Effect of tetrandrine on intraacinar pulmonary arterial structural remodelling and pulmonary hypertension]. AB - Rat's pulmonary hypertension was induced by a single subcutaneous injection of monocrotaline. The influences of tetrandrine (Tet) upon the structure of intraacinar pulmonary arteries (IAPA) was studied by light and electron microscopy. The results indicated that Tet reduced the thickness of IAPA medial membrane, the ratio of the thickness of IAPA medial membrane to the diameter of IAPA, the number of circular muscular artery and partially muscular artery about 16.4%, 30.5%, 24.3% and 16.4% respectively, and Tet increased the number of nonmuscular artery about 24.7%. It also decreased the degeneration of IAPA, collagen of medial membrane, prolife-ration and myoid differentiation of the pericyte, and it increased the number of IAPA. The authors discussed why Tet caused such changes. PMID- 9208621 TI - [The preparation of hepatic receptor imaging agent 99mTc-DTPA-NGA]. AB - The method of preparation and the character of the hepatic receptor imaging agent 99mTc-DTPA-NGA have been presented. The radio-chemical purity of the imaging agent is higher than 95% and the stability of the imaging agent is more than 6 hours in vitro and in human blood. The agent is able to tend to liver and has the character of receptor-intervene-conduct binding. Our comparative study of 99mTc DTPA-NGA versus 99mTc-NGA reveals, the stability of the former in vitro and in human blood is better than that of the latter. The imaging in rabbits shows that the imaging quality of the former is higher than that of the latter. The radioactive metabolites in urine have been examined. The former does not fall off 99mTc in vivo, but the latter falls off 99mTc clearly. 10 clinical hepatic imagings have had satisfactory results. PMID- 9208624 TI - [Protective and treated effects of L-arginine on hypoxia-induced pulmonary hypertension in rats]. AB - The protective and treated effects of L-Arginine on isobaric hypoxia-induced pulmonary hypertension were studied in rats. It was found that L-Arginine significantly inhibits hypoxic pulmonary hypertension and right ventricular hypertrophy. The mean pulmonary arterial pressure (mPAP) was 3.58 +/- 0.06 kPa and right ventricle/(left ventricle plus septum) ratio [RV/(LV+S)] was 0.34 +/- 0.01 in rats exposed to hypoxia (FiO2 = 0.10 +/- 0.05) for 3 weeks. The mPAP was 2.61 +/- 0.09 kPa and RV/(LV+S) was 0.26 +/- 0.01 in rats pretreated with L Arginine [200mg/(kg.d)]i there is significant difference between the two groups. We also found that L-Arginine (300mg/kg) can decrease mPAP in rats with hypoxic pulmonary hypertension. The results suggest that L-Arginine might have significant protential for protection and treatment of hypoxic pulmonary hypertension. PMID- 9208623 TI - [Analysis of 106,272 cases of postterm pregnancy in China]. AB - 106272 cases of postterm pregnancy in 945 hospitals in China during the period from Oct. 1986 to Sept. 1987 were surveyed, the overall incidence rate was 85.48/1000. Provincially, the highest two rates were 127.36/1000 and 120.92/1000 in Jilin and Shandong provinces respectively, and the lowest two rates 53.98/1000 and 61.38/1000 in Xizang and Qinghai provinces respectively. The perinatal mortality was 27.90/1000 (corrected 12.49/1000). The incidence of macrosomia (89.50/1000) in postterm pregnancy was twice as much as that in term delivery. Postterm pregnancy was more frequent in pregnant women of 20-34 years old,and of 42 weeks' gestation. The incidence malformations in postterm pregnancy was 154.13 per ten thousand, in which the frequency of malformations associated with low birth weight was three times that associated with macrosomia. PMID- 9208625 TI - [Relationship between the express product of src gene (pp60c-src) and the initiation of gastric carcinoma]. AB - The tissues of the fetal gastric epithelia, adult normal gastric epithelia, gastric epithelial lesions, gastric carcinoma and the tissues adjacent to carcinoma were studied immunohistochemically by using the specific monoclonal antibody of c-src gene express product (pp60c-src), MAb 327. The positive rates of pp60c-src in the fetal gastric epithelia, adult gastric epithelial inflammatory hyperplasia, intestinal glandular metaplastia and gastric carcinoma were 100.0% (12/12), 100.0% (30/30), 100.0% (16/16), 83.3% (40/48), respectively, all these pp60c-src positive rates were higher than that of adult normal gastric epithelia (60.0%, 12/20), a significant difference was noted statistically (P < 0.005). And there was statistical significant difference of pp60c-src express amount among different tissues (P < 0.005). The positive rates of pp60c-src in the intestinal and diffuse type gastric carcinomans were 100.0% (16/16) and 68.8% (22/32), respectively, a significant difference was found statistically (P < 0.025), the difference of pp60c-src express amount was significant (P < 0.05) but no significant differences of the positive rates and express amount of pp60c-src in the tissues adjacent to carcinoma of two types of carcinoma were found (P > 0.05). The results of this experiment indicated that the activation of c-src gene and increase of pp60c-src express were associated with the proliferation, turnover, transformation and malignant change of the gastric epithelial cells, with the initiation of the gastric carcinoma, and also with the differentiation and the histological types of the gastric carcinoma. PMID- 9208626 TI - [Effect of methimazole and dexamethasone on leucocyte glucocorticoid receptor, plasma ACTH, and cortisol levels in Graves' disease]. AB - Thirty-two cases of newly diagnosed Graves' disease with hyperthyroidism were recruited in this study on the changes of leucocyte glucocorticoid receptor (GCR), plasma ACTH, and cortisol levels befor and after treatment with methimazole (tapazole) alone (n = 16) and methimazole combined with Dexamethasone (Dex, TD group, n = 16). Twenty normals served as the control. Methimazole treatment was initiated with a dosage of 40 mg/d, tapered to 20 mg/d after two weeks and maintained until complete remission in both groups. Meanwhile, Dex 6 mg/d was added to the TD group along with methimazole from the commencement of therapy. The dosage of Dex was reduced to 4.5 mg/d on the 5th day and to 2.25 mg/d two weeks after treatment, and continued until remission. It was found that there was remarkable decrease in leucocyte GCR levels with a moderate elevation of plasma ACTH and a slight decline of plasma cortisol in untreated Graves' disease, suggestive of a compensation of the pituitary-adrenal axis function in hyperthyroidism. The levels of GCR, ACTH and cortisol returned to normal after complete remission by methimazole in the methimazole alone group. In the TD group, however, all GCR, ACTH and cortisol levels were significantly decreased after Dex therapy, implying down regulation of GCR by exogenous Dex and suppresion of the pituitary-adrenal axis. Thus the dosage of glucocorticoid should be appropriately adjusted to avoid adrenal insufficiency, especially in case of stress, due to the suppresion of pituitary adrenal function. PMID- 9208627 TI - [Measurement of maximal inspiratory flow and forced inspiratory capacity and its clinical application]. AB - The methods of measuring the maximal inspiratory flow (V(imax)) and the forced inspiratory capacity (FIC) via forced inspiratory capacity-time curve (FIC-t curve) were investigated. Both V(imax) and FIC were measured in 35 normal subjects and 89 patients with chronic obstructive pulmonary disease (COPD). The maximal inspiratory pressure (MIP) was measured simultaneously at function residual capacity(FRC) level by modified Black Method. The results showed that there is a linear relationship between MIP and V(imax) or FIC in both normal subjects and patients with COPD. Normal subjects had a mean V(imax) and FIC much higher than those of patients with COPD. The values of V(imax) and FIC in patients were also, significantly correlated to the severity of COPD. So we suggest that both V(imax) and FIC be used as clinical indices to reflect inspiratory muscles strength. PMID- 9208628 TI - [Curative effect of Interfon-Alpha in children with infectious mononucleosis]. AB - Thirty-one cases of infectious mononucleosis treated with Interfon-Alpha were reported. The dose of intramuscular injection was one million units per day for 5 7 days. The recovery course of fever, angina, lymphadenopathy and hepatosplenomegaly was much shorter in the study group than in the control group (27 cases). The results suggested that interfon-Alpha should be efficacious against EBV activity and might shorten the course of this disease. PMID- 9208629 TI - [The bonding strength between plat castable ceramics and mild fusing alloy]. AB - Plat castable ceramics (PCC) was bonded to SDA-I mild fusing alloy using three bonding agents, and the tensile bond strengths were tested respectively in order to select the best bonding agent and the best way of surface treatment. The results obtained with scanning electron microscope and X-ray energy spectrum showed that the bonding behavior of TF agent was the best one. The greatest tensille bond strength (24.37 MPa) was obtained with TF agent when the surface of PCC was etched by hydrofluoric acid and the alloy was painted with KH-570. Remarkable tensile bond strengths were noted in the group where EM agent was used with PCC etched by hydrofluoric acid and alloy treated with sandblasting (15.20 MPa), and in the group where Porcelite dual cure cement was applied with PCC painted by KH-570 and alloy treated with sandblasting (15.25 MPa). PMID- 9208632 TI - [Effect of birchdust on viability of alveolar macrophages and superoxide produced by alveolar macrophages in vitro]. AB - Rabbit alveolar macrophages (AM) in vitro were used as a model in this wooddust toxicological experiment to study the effect of birchdust with different concentrations and exposure durations on the viability of AM and superoxide produced by AM. The results showed that the viability rates in three birchdust groups (400, 800, 1600 micrograms/ml) gradually decreased when the concentration at all the designed exposure durations (12, 18, 24 h) increased except the rates for the 6-hour period. On the contrary, the contents of superoxide (O2-.) at the first two durations (6, 12 h) increased rapidly and reached their highest level at the 18 hours. Moreover, the rapid increase of O2-. preceded the declination of viability rates, suggesting that birchdust toxicity in AM in vitro be probably related to dust-induced O2-. produced by AM. PMID- 9208631 TI - [A 5-year before-and-after comparison of lung function in asbestos workers]. AB - A 5-year before-and-after comparison of lung function was carried out in 119 male asbestos workers. Of them 50 were healthy workers (0), 25 were patients with suspectable asbestosis (0+), 36 with Stage I asbestosis (Ias) and 8 with Stage II asbestosis (II as). The findings measured for before and after 5 years revealed that FVC, FEV and DLco declined in the four groups over the whole period of observation. The decrease in FVC and DLco was most marked and the differences were statistically significant (P < 0.05). Comparison among four groups showed that all indices of lung function in Group II as reduced significantly after 5 years (P < 0.01), indicating a rapid decrease of lung function in Group II as. It is suggested that FVC and DLco are the most sensitive indices in detecting very early abnormalities of lung function in asbestos workers. PMID- 9208630 TI - [Methodology for controlling healthy worker effect on coal miners' mortality]. AB - This paper reports how to control the healthy worker effect (HWE) in a mortality study of coal miners. We used four methods, namely (1) age-specific corrective coefficient, (2) proportional mortality ratio (PMR), (3) corrective standarized mortality ratio (CSMR), and (4) a control group composed of factory workers. The results showed that all these methods could control HWE, and the total mortality in the coal miner group was significantly higher than that of the general population group (P < 0.05). However, the best way for controlling HWE awaits further studies. PMID- 9208633 TI - [Isolation and structure flucidation of alkaloids from the bulb of Fritillaria Wabuensis S.Y. Tang et S.C. Yueh]. AB - Eight alkaloids were isolated from the bulb of Fritillaria Wabuensis S.Y. Tang et S.C. Yueh the best "Chuan Bei" cultivated in Sichuan province, which is efficacious against cough. Five of them are known, identified as imperialine (I), peimisine (II), ebeinone (III), isoverticine (IV), and imperialine-beta-N-oxide (V). All the signals for both 1H and 12C-NMR spectra of base (V) were assigned by 2D NMR experiments, and an X-ray structural data of alkaloid (V) was obtained for the first time. PMID- 9208634 TI - [The colorimetric method for measuring activities of lipoprotein lipase and hepatic lipase in plasma]. AB - To study the pathogenesis of hyperlipoidemia and atheromatosis and the metabolism of lipoprotein, we have developed a colorimetric method for simultaneously determining the activities of post-heparinplasma lipoprotein lipase (LPL) and hepatic lipase (HL). The intralipid was kept for LPL and HL at 37 degrees C, pH8.3 for 30 min, with 100 microliters post-heparin plasma. The LPL and HL in the post-heparin plasma could hydrolyse the triglyceride in intralipid into glycerine and free fatty acid (FFA). Determining the amount of FFA by copper-reagent method, we could measure the activities of LPL and HL. The kinetics of LPL and HL in post-heparin plasma was observed. K(m) values for LPL and HL were 0.9 mumol/L and 2.4 mumol/L respectively. The C. V. for LPL and HL were 4.5% (n = 4), 2.9% (n = 6) and 6.4% (n = 6), 4.8% (n = 6) respectively. PMID- 9208635 TI - [Labeling DNA probe by polymerase chain reaction]. AB - The use of polymerase chain reaction (PCR) for labeling probe has been demonstrated to offer various advantages including efficient labeling of DNA fragments as small as 72 bp, direct labeling of genomic DNA, and labeling with subnanogram amounts of input DNA. Therefore, a procedure for the nonradioactive labeling of chromasomal DNA 203 bp fragments of Helicobacter pylori with the hapten digoxigenin (Dig) by PCR has been developed. The results showed that the concentration of probe labeled by PCR was much higher than that by random primer labeled. PCR has the advantage of rapidity and economy. It is a very effective technique for synthesis of Dig-labeled DNA probe. PMID- 9208636 TI - [Determination of metronidazole in serum by HPLC]. AB - This paper report a sensitive and rapid method for the determination of metronidazole (MTZ) using theophylline as the internal standard. High performance liquid chromatograph model 344 (Beckman) with a 254 nm wavelength UV detector and YWG-C18H37 column (10 microns, 250 x 4.6 mm) was used. To the serum sample 200 microliters, 100 microliters phosphate buffer (0.8 mol/L, pH 7.5) was added, then extracted with 3 ml chloroform containing 5% isopropyl alcohol. The organic layer was removed and evaporated to dryness under an air stream in a 40 degrees C water bath. The residue was dissolved in 30 microliters mobile phase and 20 microliters injected. The mobile phase of water-methanol (73:27) was pumped at 1.0 ml/min through the column. The detector operated at 0.005 aufs. The retention times for MTZ and theophylline were 5.78 and 6.81 min respectively. Standard curve was linear in the concentration range of 0.3125 to 20 mg/L. The detection limit in serum was 0.02 mg/L. Extraction recovery was 77%-82%; method recovery 99%-102%; withinday RSD less than 3.0%; inter-day RSD less than 3.5%. PMID- 9208637 TI - [Study on the synaptonemal complexes in spermatocytes of the brown rat (Rattus norvegicus caraco). II. Study on the behavior and morphology of the sex chromosomes pairing]. AB - The behavior and morphology of sex chromosomes pairing in spermatocytes of the brown rat have been studied by surface spreading, silver and phosphotungstic acid(PTA) staining techniques. The results are as follows. Sex chromosomes axial cores thicken before X and Y pairing. X Y pairing is delayed until early pachytene. The first pairing initiation site is located on telomere of the short arms of X and Y chromosomes. The second pairing initiation site is located on teltomere of the long arms of X and Y chromosomes or on the interstitial position of the long arms of X and Y. Pycnosis speed of sex axial cores is different with different stages during prophase I of meiosis. In middle Pachytene the almost whole Y is fully paired in SC association with about one-third the X. On the region of X-YSC, lateral elements of X-YSC divide into two skeins, bubble appears on one of the two skeins and the unpaired X and Y axial cores are in variable forms. X Y pairing initiation sites, relationship of X Y pairing and genetic homology and mechanism of thickening and variable X Y axial cores were discussed. PMID- 9208638 TI - [Studies on the phenotype deviation of the single-cell subclones of human stomach carcinoma MGC-803 cell line]. AB - The diversity of phenotypes of tumor cells is the main reason for the differences of drug-sensitivity, immunology, metastatic properties and cellular growth rate, and thus raises difficulties in tumor diagnosis and therapy. In order to investigate the mechanisms involved in the expression, correlation, development and regulation of the diversed malignant phenotypes, cell models from single-cell subclones of human stomach carcinoma cell line MGC-803 were studied. According to the growing time needed for the process of separation of each single-cell subclone, they were categoried into 3 groups as fast growing, moderatly and slowly growing subclones and named. From each group, representative subclones were selected--M17 fast. M6 moderate and M3 slow, for further comparative studies. Foci frequency in soft agar was very high with M17, very low with M3, and M6, the middle. The difference in the major transformed phenotype among the 3 subclones were statistically significant. There were differences in the function of gap junctional intercellular communication as detected by scrape-loading and dye transfer method. M17 negative, while M6 and M3 were positive with dye transfer. Immunocytochemical staining of cytoskeleton elements showed heavy disruption of microtubule and microfilament networks and appearance of F-actin aggregates in M17 cells. On the other hand, M3 showed less disruptive changes and more normal appearance of microtubules and microfilament organization. Immunofluorescent staining showed marked differences in gene expression of c-myc, c-Ha-ras and c-met between M17 and M3. M17 expressed intense fluorescence of the 3 onco-proteins while M3 was negative with c-myc and c-Ha-ras, only weak c-met fluorescence in the cytoplasm. PMID- 9208639 TI - [Analysis of chemokine (S) produced by mouse thymic stromal cell lines]. AB - By using Boyden Chamber and PACS methods, we analysed the chemotactic activity of five mouse thymic stromal cell (MTSC) lines' culture supernatant for neutrophil, monocyte/macrophage and lymphocyte, and the phenotype of the attracted lymphocyes, MTEC 1, MTEC 2, MTEC 3, MTEC 5 are mouse thymus epithelial cell lines. MTDC 4 is a mouse thymus dendritic cell line. The results indicate that all of the five MTSC supernatants have chemotactic activity for the target cells mentioned above but in different degree. Chemokines produced by the MTSC cells can be classified into three categories: (1) Chemokines produced by MTEC 1 and MTEC 2 are more potent for the attraction of neutrophils and lymphocytes. (2) MTDC 4 derived chemokines are highly potent for the attraction of monocytes/macrophages. (3) Chemokines produced by MTEC 3 and MTEC 5 are equally attractive to neutrophils, lymphocytes and monocytes/macrophages. Most of the MTSC-SN are more potent for the migration of B lymphocyte than for T lymphocyte. The MTSC-SN exhibits higher chemotactic activity to CD4-CD8+ T cell subset than to CD4+CD8-T cell subset. The characterization of chemokines in MTSC-SN will benefit the finding of new chemokines and the analysis of the mechanism of thymus homing. PMID- 9208640 TI - [The construction of 2 BS cell lines overexpressing PKC beta I isoform and the priliminary analysis of the relationship between expression of PKC beta I and cell proliferation]. AB - Using gene transfection technique, we have constructed a series of human embryonic lung cell lines that overexpress stably a full length cDNA encoding the beta I isoform of PKC for the first time. BS-PKC 3 is a cell line containing about 3 fold greater PKC activity than parental cell lines or control cells that carry an integrated vector lacking the cDNA inset. In comparison with control cells and parental cells, these cells exhibit significantly enhanced growth rate. The expression of oncogene c-myc which is relating closely to cell proliferation is also increased obviously in BS-PKC 3 cell. We have firstly evidence that the overexpression PKC beta I affect the level of the expression of c-myc in the 2 BS cells. It seems that there may be one of the molecular mechanisms of the effect on the proliferation in the 2 BS cells by PKC beta I. PMID- 9208641 TI - [CD4-CD8- to CD4-CD8+ transition induced by anti-CD3 mAb in vitro cell culture system]. AB - In our experiments, we have observed the effect of anti-CD3 mAb in inducing the differentiation of CD4-CD8- (double negative, DN) thymocytes into CD4+ CD8+ (double positive, DP) cells in an in vitro cell culture system including IL-7 for maintaining the growth of TN thymocytes. When TN thymocytes were stimulated by immobilized anti-CD3 mAb for 3 days, CD4-CD8+ cells generated, which were mostly immediate precursors of CD4+CD8+ thymocytes according to their surface TCR beta expression. The transition from DN to DP thymocytes accompanied with downregulation of interleukin 2 receptor (IL-2 R) alpha chain, and the TCR alpha beta-CD3 expression induced by IL-7 was also inhibited. Anti-CD3 mAb was effective only before the appearance of functional TCR-alpha beta-CD3 complex. Taking together, the results strongly suggest that anti-CD3 mAb induced DN thymocyte defferentiation is through the pre-TCR complex. PMID- 9208643 TI - [A new way for inbred strain mice genetic monitoring and the discovery of sex linkaging RAPD markers]. AB - We used 21 10 bp random primers to amplify DNA for four colonies of BALB/ c mice, four individuals of C 57 BL mice, and four individuals of Kunming stock mice. The amplified band patterns were different between BALB/c and C 57 BL mice in the products of 13 primers, and 8 primer products showed difference between BALB/c and Kunming stock mice. These results indicated that we can easily distinguish different strains of mice by RAPD method. For the four colonies of BALB/c mice, the genetic background of Chengdu colony. Shanghai colony and Beijing colony were homogeneous at all RAPD markers, however, in Kunming colony, there were two BALB/c individuals were disclosed different amplified patterns by 4 primers, showing that these two Kunming BALB/c mice maybe suffer genetic contamination or mutation some time. An unexpected phenomenon discovered in this study is that sex specific amplification bands were amplified by primer OPG 2, OPE 4, OPE 9 in all male mice. Although an 1.2 kb band were disclosed by both OPG 2 and OPE 4 in male mice, cross RAPD and not hybridization showed that the two bands were not homologous. PMID- 9208642 TI - [The diversity of human hematopoietic stem/progenitor cells. I. Two-color flow cytometric analysis of the different functional subpopulations of CD 34+ hematopoietic stem/progenitor cells riched from human bone marrow]. AB - In hematopoiesis; the human CD 34 protein is a strict developmental stage specific antigen that marks hematopoietic stem/progenitor cells, suggesting that it plays an essential role in hematopoiesis. More recently, it has been demonstrated that hematopoietic cells expressing the CD 34 antigen constitute various heterogeneous cell populations in which each CD 34+ subset was associated with commitment to a particular lineage, and differed from other's in reconstituting hematopoiesis. In this report, we have assessed the different functional subpopulation of CD 34+ hematopoietic stem/progenitor enriched from human bone marrow using Isolex TM 50 system according to the strategy on immunomagnetic separation of positive selection. CD 34+ cell population of high purity (> 90% CD 34+) was analyzed by means of double staining procedure of flurorescein conjugated monoclonal antibodies on the two-color FACAcan or FACS 440. Eight cell subsets of at least of bone marrow CD 34 hematopoietic stem/progenitor cells have been defined by undertaking a comparative coexpressing of CD 71, CD45, CD 33 and HLA-DR antigens on CD 34 hematopoietic stem/progenitor cells. The frequencies of various subsets in two illustrative are as following: 1). CD 34+/CD 71- and CD 34+/CD 71+ (23.43%-56.6% versus 33.4%-66.6%): 2). CD 34+/CD45- and CD 34+/CD45+ (80.8%-82.5% versus 8.1%-11.2%): 3). CD 34+/CD 33- and CD 34+/CD 33+ (20.4%-80.6% versus 14.6%-64.8%): 4). CD 34+/DR- and CD 34+/DR+ (6.3%-11.0% verus 82.8%-85.5%). Immunological double color staining of IGSS-APAAP was also used to further analyse the CD 34+ cell subsets as above-mentioned, the results were very similar to those obtained by FACScan. Our data indicate that CD 34+ hematopoietic cell fraction is far from being a uniform cell population, and many works regarding biological properties and regulation mechanism of different subsets of CD 34+ hematopoietic stem/progenitor cells are being carried out. PMID- 9208644 TI - [Expression of cDNA of human chorionic gonadotropin beta-subunit (beta-hCG) cDNA in insect cells and effect of expressed product on mouse lymphocytes in vitro]. AB - Expression vector pVL 1393-hCG beta containing beta-hCG cDNA has been constructed using an unfused protein nuclear polyhedrosis virus (AcNPV) expression vector. The insect cells (Sf 9) were cotransfected by the expression vector and nuclear polyhedrosis virus genomic DNA, and recombinant virus AcNPV-hCG beta was screened out, beta-hCG cDNA was expressed in insect cells infected by recombinant virus and recombinant beta-hCG (r beta-hCG) was secreted into medium. The purity of r beta-hCG, purified by immuno-affinity chromatography, was about 90% and the molecular weight of r beta-hCG was 22,500 Da. Like hCG, r beta-hCG suppressed significantly proliferation of induced lymphocytes, as well as production of IL-2 to some extent, on a parallel with suppression of lymphoproliferation. PMID- 9208645 TI - [A model of osteoporosis induced by retinoic acid in male Wistar rats]. AB - An animal model of osteoporosis induced by retinoic acid was successfully established in 3-month-old male Wistar rats. The animals were given the drug 70 mg.kg-1.d-1 for 14 d intragastrically and sacrificed on day 29. The proximal tibia and middle tibia of the rats were processed undecalcifiedly for quantitative bone histomorphometry. Compared with the control rats, the cancellous and compact bone volume of the model rats were reduced markedly. The bone tissue microstructure showed some obvious pathological changes that the trabecular number were decreased, the separation of trabecular and medulla ossium cavity became large, the thickness of trabecular and cortex of bone were decreased. The mechanism of bone loss in the model rats was that the osteoclast was activated by retinoic acid which promoted bone resorption. Other changes in the model rats were also observed such as: the body weight, and the weights of seminal vesicle and prostate were decreased, the adrenal glands and spleen showed hyperplasia and hypertrophy. No change of the blood concentration of calcium, phosphorus, alkaline phosphatase, alanine transaminase, estradiol and testosterone in the model rats was observed. PMID- 9208646 TI - [Effect of dl-3-butylphthalide on the striatum extracellular amino acid and dopamine contents in the rat during cerebral ischemia]. AB - The effect of dl-3-butylphthalide (NBP) on the contents of amino acids and dopamine in the rat striatum during globe cerebral ischemia has been studied. By using the technique of microperfusion in the striatum of rats subjected to 4 vessel occlusion cerebral ischemia, the extracellular contents of glutamate, taurine, gamma-aminobutyric acid and dopamine were found to be significantly increased in the striatum during the 20 min of cerebral ischemia. NBP (40 mg.kg 1; i.p. 30 min before ischemia) was shown to reduce the contents of dopamine and its metabolite, 3,4-dihydroxyphenylacetic acid (DOPAC), in the striatal extracellular fluid of the rat during ischemia. The content of glycine before and after ischemia was also reduced. However, no significant effect on the contents of glutamate and some other amino acids was observed. The results suggest that NBP may improve the striatal ischemic injury. PMID- 9208647 TI - [Effect of thyrotropin-releasing hormone analogue (YM14673) on experimental brain contusion and edema in rats]. AB - The effect of TRH analogue, YM14673: N alpha-[[(S)-4-oxo-2-azetidinyl]-carbonyl) L-histidyl-L-prolinamide] dihydrate, on traumatic brain edema were studied in male Wistar rats. The cerebral contusion was produced by Feeney's dropping weight method. Animals were randomly divided into 4 groups: normal group and YM14673 (0.1 mg.kg-1, i.p.), YM14673 (1.0 mg.kg-1, i.p.) and a control group (given equal volume of physiological saline, i.p.) after cerebral contusion. Contents of water in brain tissue were measured. These results suggest that treatments with YM14673 significantly reduced brain edema than control group (P < 0.05). PMID- 9208648 TI - [Pharmacokinetics of naltrexone hydrochloride and naltrexone glucuronide in the dog]. AB - Pharmacokinetics of naltrexone hydrochloride (NTX) and naltrexone glucuronide was studied in the dog using HPLC-electrochemical detection with naloxone as internal standard. After iv 5 mg or po 10 mg NTX, the plasma concentration-time curves of NTX were found to fit to a two-compartment model and a single compartment with first-order absorption. The elimination half-lives of NTX were 78 +/- 6 min and 74 +/- 6 min, respectively. Although NTX could be absorbed rapidly in the dog after po administration, the plasma concentration of the parent drug was very low and its absolute bioavailability was 15.8%. The experiments showed that the major metabolite of NTX in dog plasma was beta-glucuronidase-hydrolyzable conjugate. Dosing NTX intravenously and orally, the plasma levels of the conjugate were 1.3 and 23 times as high as that of the parent drug, the elimination half-lives of the glucuronide from plasma were 3.4 h and 12.6 h, respectively. The results indicate that NTX is subjected to a marked first-pass effect in the dog after oral administration. PMID- 9208649 TI - [Systematic analysis of basic drugs in plasma using X-5 solid-phase extraction GC FID and GC-MS]. AB - A systematic determination method for 34 basic drugs in human plasma is described. Drugs were extracted from plasma at pH 10 using X-5 resin as adsorbent, then identified and quantitated by capillary GC-FID and GC-MS. The detection limits for most drugs are in the range of 0.5-2.0 micrograms.ml-1. Precision and linearity of the method are satisfactory for clinical toxicological applications. PMID- 9208650 TI - [Studies on preparation and characteristics of cisplatin chitosan microspheres]. AB - In this paper, the preparation, drug content, size and size distribution, appearance and morphology, release characteristics in vitro and degradation characteristics of cisplatin chitosan microspheres (CDDP-DAC-MS) were studied. CDDP-DAC-MS were prepared by emulsion-crosslink technique. The CDDP-DAC-MS was shown to have rough spherical surface under scanning electron microscopy. The average diameter of the microspheres was 74.80 microns and CDDP content was 20.83% +/- 0.36%. CDDP-DAC-MS swelled slightly in saline after 1 h. Within the test period, the release of CDDP from CDDP-DAC-MS in saline solution could be described by first-order equation. The microspheres were sterilized by 60Co radiation. After 28 d of hepatic artery embolization with CDDP-DAC-MS in dogs, pathological photomicrograph showed that CDDP-DAC-MS could still be observed. PMID- 9208652 TI - [Animal models of osteoporosis: status and prospect]. PMID- 9208651 TI - [The antifertility effect of gossypol plus testosterone and estrogen]. AB - In order to reduce the side effect of gossypol, gossypol was used in combination with steroid hormones so that the dose of both drugs can be reduced. Silastic capsules containing testosterone (T) + estradiol (E) were implanted under the skin male wister rats for 8 weeks. After removing the implants, testosterone was given orally at the dose of 15 mg.kg-1 which is only 50% of the usual antifertility dose. Mating tests showed that the male rats became infertile. Microscopic examination of the heart, liver and kidneys showed no pathologic changes. The treated rats gained body weight as well as the controls. The fertility of the treated rats recovered four to five weeks after treatment. Thus, gossypol in combination with testosterone and estrogen exhibited a low degree of side effect and high antifertility activity. PMID- 9208653 TI - [A study on the viability of corneal endothelium in rabbit by dual staining with trypan blue and alizanin red S]. AB - PURPOSE: We study the viability of preserved rabbit corneal endothelium by dual staining with trypan blue and alizanin red S. METHOD: We randomly divided New Zealand white rabbit eyes into four groups, each with 5 eyes. The eye balls were preserved in moist chambers with aqueous remorad and C3F8 temponade for 5, 7, 10 and 14 days. Then we got the corneal Bottons with sclera ring. Isolated corneas were placed with the endothelial side up in a corneal cup, and trypan blue (0.25%) was added drop-wise to cover the endothelium. After 1.5 minutes the stain was poured out. The endothelial layer was then covered with aliznin red S (0.2%) for 1.5 minutes and pouring away the staining reagent. After staining, the corneas were immensec in glutaraldehyde fixative solution for 10 minutes. Finally, the endothelium was examinated and taken photoes with light microscope. RESULTS: Changed cells that have become permeable to trypan blue show deep blue staining of their nuclei in contrast to the unstained condition of normal cells. All the intercellular borders of endothelial cells were purple. The survival rate of the endothelium preserved for 5 and 7 day is more than 95%, for 10 days is 75 80%, and for 14 days about 70%. CONCLUSION: This technique allows a proportion cout of damaged and undamaged cells in the endothelial monolayer, makes it possible to observe their structures clearly and gives advantage to take pathelage photos. PMID- 9208654 TI - [Management of the dry eye with parotid duct transplantation: a summary on 40 cases]. AB - PURPOSE: As today the effect of parotid duct transplantation is clinically regarded as uncertainty, and there is still absent of effective managenent of the dry eye, this paper tries to clarify its value, and see if it can be used clinically. METHODS: Forty dry eyes were operated with a follow up of 3 weeks to more than 6 years in the past 42 years. The pre- and postoperative changes of tear flow and vision, and the cause of dry eye, operative method, length of parotid duct and complications of bilateral operation were briefly summarized. RESULTS: Of these 40 operated eyes postoperatively, 82.5% had tearing, but no tearing occurred in 27.5%. Finally: vision increased in 72.5% with out any decrease. There was no difference in the results whether the procedure done extraorally or intraorally. The parotid duct being shorter was only seen in 7.5%, which were satisfactorily managed by tube making with oral mucosa. Four cases operated binocularly and silmutaneously, only 1 occurred aveolar abscess with complaining dry mouth, but cured 1 week later. After operation, profusive tearing occurred in 25 dry eyes caused by trachoma during eating and 96% usually, but tearing stopped finally in 3 eyes (2, due to infection). Four dry eyes with ocular pemphigoid failed. Among 9 dry eyes resulted from Stevens-Johnson syndrome, 6 had little tearing postoperatively, 2 of which attained a nearly normal results. One of the two dry eyes resulted from alkali eye burn failed and the other, occurred tearing but only followed-up for 1 month, the result was uncertain. CONCLUSION: As the technique of the parotid duct transplantation is simple and easy with no much complications, and for treating the dry eye, there is up to now still absent of effective management, therefore, the parotid duct transplantation should be considered to be indicated in dry eyes esp, caused by Stevens-Johnson syndrome. Though its success rate is lower than that of traucomatous ones, the satisfactory result of nearly normal postoperative tearing may be got. In ocular pemphigoid, however, it is contraindicated. PMID- 9208655 TI - [The relations of corneal, lenticular and total astigmatism]. AB - PURPOSE: To determine the relations of corneal, lenticular and total astigmatism and the changes of the astigmatism with age. METHOD: Out-patients with refractive errors were refracted with retinoscope after using cycloplegic drops and measured the radii of anterior corneal curvature. RESULT: One hundred and ninety-four cases (382 eyes) with refractive errors were studied. Of the eyes 67.9% had regular corneal astigmatism, 68.1% irregular lenticular astigmatism and 60.7% regular total astigmatism, 88.5% of the corneal astigmatism has the same quality as the total astigmatism. The total astigmatism in 46% of the eyes included the summation of corneal and lenticular astigmatism, but in 41.3% of the eyes irregular lenticular astigmatism corrected the regular corneal astigmatism. The astigmatism of cornea, lens and total astigmatism changed from regular to irregular with the increase of age. The linear correlation analysis showed a positive correlation between the power of horizontal corneal refraction and age, and a negative corrlation between the power of vertical corneal refraction and age. CONCLUSION: The shape of cornea was the major cause of total astigmatism. The influence of lens on the total astigmatism was different. The reasons for the change of the total astigmatism from regular to irregular with the increase of age were the changes of the power of corneal refraction, particularly the increase of the power of horizontal corneal refraction and lenticular irregular astigmatism. PMID- 9208656 TI - [The study of correlation between hemorrheology and fluorescein angiography in open-angle glaucoma]. AB - PURPOSE: To observe the correlation between hemorrheology and arm-retinal filling time of fluorescein angiogaphy in patients with primary open-angle glaucoma. METHODS: The whole blood apparent viscosity in high (125 S-1), moderate (23 S-1) and low (1.2435 S-1) shear rates, plasma viscosity and hematocrit were measured in 50 cases (50 eyes) with primary open-angle glaucoma whose intraocular pressures were controlled. They also underwent fluorescein angiography. The duration from arm to retina arteria filling was named arm-retinal arteria filling time. RESULTS: There were positive correlation between the whole blood apparent viscosity in high and moderate shear rates, hematocrit and arm-retinal arteria filling time (P < 0.05-0.005). The higher blood viscosity, the longer arm-retinal arteria filling time. CONCLUSION: The blood viscosity in patients with primary open-angle glaucoma can affect the arm-retinal arteria filling time of fluorescein angiography. So, the increasing blood viscosity in patients with primary open-angle glaucoma can reduce the blood supply to the optic disc. PMID- 9208657 TI - [An experimental study on homoharringtonine and glaucoma surgery]. AB - PURPOSE: To test the inhibiting effect of a Chinese herbal drug. Homoharringtonine (HH) on the wound healing process at the filtering site in an experimental model. METHODS: Posterior sclerectomies were performed in 20 rabbits. Postoperatively one eye of each rabbit received subconjunctival injections of HH and fellow eye received saline injections in a randomized masked fashion. RESULTS: 14 days after operation, intraocular pressure decreased significantly compared with that of control (P < 0.05). The number of filtration blebs was greater than that of control (14/6). On pathological examination, the number of fibroblasts per square measure of cross section of tissue cut from filtration region that was diminished significantly compared with that of control (P < 0.01). There were no serious and permanent ocular toxic and side effects. CONCLUSIONS: The study suggests that the HH can inhibit cicatrization at the glaucoma filtering site, promote the formation of filtering blebs and provide a sufficient evidence for the further clinical use of HH. PMID- 9208658 TI - [The study of grading standard photos of oblique flashlight test]. AB - PURPOSE: We try to establish a convenient, fast, inexpensive and accurate screening test and to make a grading standard. METHODS: 1. In 120 eyes, we measured axial anterior chamber depth (AACD), peripheral anterior chamber depth (PACD), antrior chamber angle (ACA) and took iris pictures under slitlamp microscope imitating the oblique flashlight test. The clear photos from 107 eyes were measured by computerized image analysis to determin the ratio of the width of nasal iris and nasal iris light band (ILBR). Then the linear correlation of ILBR with AACD, PACD and ACA was analyzed statistically. The ILBR was graded according to the analytic results. RESULTS: 1. Relationship of ILBR with PACD, AACD and ACA: the linear correlation analysis showed a positive correlation relationship among ILBR, PACD and AACD (r = 0.9451, P = 0.0001 and r = 0.8725, P = 0.0001), and showed a negative correlation relationship between ILBR and ACA (r = -0.7582, P = 0.0001). 2. ILBR grading standards: grade 1. ILBR < or = 1/5, the gonioscopy usually demonstrates a dangerously narrowed angle (N3-4); grade 2, ILBR > 1/5- < or = 1/4, the gonioscopy will always show a capable of closure angle (N2-3); grade 3, ILBR > 1/4- < or = 1/3, that angle is incapable of closure (N1-2); grade 4, ILBR > 1/3- < or = 1/2, the most angles show wide (W-N1); grade 5, ILBR > 1/2, the gonioscopy shows all wide angle. The linear correlation analysis also showed correlation relationship among ILBR grades PACD, AACD and ACA. CONCLUSION: Through arational ILBR grades and a set of grading standard photos, it is found that the test is not only convenient, fast, and inexpensive, but also accurate as well. The test not only plays an important role in PACG screening, but also may be applied widely to estimate shallow AACD and narrow angle in the clinic. PMID- 9208659 TI - [Evaluation of accuracy of measuring intraocular pressure by handheld non-contact applanation tonometer]. AB - PURPOSE: To evaluate the accuracy of measuring intraocular pressure by handheld non-contact applanation tonometer. METHOD: 58 patients' (113 eyes) intraocular pressure were measured by Keeler, non-contact tonometer and R 900 Goldmann applanation tonometer and the results of measurement of intraocular pressure by the two kinds of tonometers were compared. RESULT: The mean intraocular pressure measured by non-contact is 16.31 +/- 5.59 mmHg and 17.49 +/- 6.13 mmHg (1 mmHg = 0.1333 kPa) by Goldmann applanation tonometer, respectively. There was no statistical significance to be found (P > 0.05) between the two methods. By linear correlation and regression analysis, a positive correlation was found between the two methods (r = 0.8942, b = 0.8154). CONCLUSION: The handheld non contact tonometer has the same accuracy and reliability of measurement of intraocular pressure comparing with Goldmann applanation tonometer, and it can be used in glaucoma clinic and screening. PMID- 9208660 TI - [Posterior chamber intraocular lens implantation in filtered glaucoma eyes]. AB - PURPOSE: To investigate the method of cataract extraction with posterior chamber intraocular lens implantation in eyes that had undergone glaucoma filtering surgery and its effect on filtering bleb. METHODS: Extracapsular cataract extraction with posterior chamber intraocular lens implantation was performed in 21 eyes with a preexisting filtration bleb, in which a lateral and inferior lateral limbal incison was selected, the pupil sphinctor was cut and the iris was sew for the small and fixed pupil. RESULTS: the postoperative visual acuity of all patients was improved in different degrees, with 0.5 or better in 76.19% of the eyes. The postoperative IOP increased by 0.41 kPa. The functional filtering bleb was not apparently cicatrized. CONCLUSIONS: Filtered glaucoma eyes with cataract can improve visual acuity and maintain the functional filtered bleb by extracapsular cataract extraction with posterior chamber intraocular lens implantation with lateral or inferior lateral limbal incision. PMID- 9208661 TI - [Transscleral suture fixation of posterior chamber intraocular lenses]. AB - PURPOSE: To determine the effects of posterior chamber intraocular lens implantation in patients with ruptured capsules of after intracapsules cataract extraction. METHODS: Using transscleral suture fixation, posterior chamber intraocular lenses were implanted in 23 eyes (23 cases), including 13 eyes after intracapsular cataract extraction and 10 eyes with ruptured capsules due to complicated extracapsular cataract extraction or traumatic cataract. RESULTS: All the patients obtained satisfactory visual acuities of 20/40 or better. Few complications were observed in the follow-up period of 3 to 16 months. CONCLUSIONS: This technique makes it possible for almost all eyes after cataract extraction to be implanted posterior chamber intraocular lens. PMID- 9208662 TI - [Reason and management of posterior capsular breaks during cataract extraction and lens implantation]. AB - PURPOSE: To study the reason and management of posterior capsular bresks during cataract extraction and lens implantation. METHODS: The authors analyse the reasons of posterior capsular brseaks during cataract extraction and lens implantation on 55 eyes and discuss the management according to the different situations. RESULTS: The posterior capsular disruption occurred most frequently during cortical cleanup on 21 eyes (38.2%) and lens implantation on 21 eyes (38.2%). The diameter of the tears was about 3 mm on 39 eyes (70.9%), about 5 mm on 13 eyes (23.6%) and larger than 6 mm on 3 eyes (5.5%). The tears with central defined borders were on 12 eyes (21.8%), with peripheral defined border in one side on 39 eyes (71%), with pooly defined borders on 3 eyes (5.5%). Final visual acuity of 0.5 or better was achieved on 35 eyes (63.6%). CONCLUSIONS: When the complication occurs during the operation, we should use the suitable management according to the different situations in order to reduce the loss of the complication. PMID- 9208663 TI - [Extraction of intraocular magnetic foreign body using intraocular earth magnet]. AB - PURPOSE: To determine the effects of extraction of intraocular magnetic foreign body with intraocular earth magnet (IOEM). METHOD: From 1992 to 1994, 22 cases with intraocular foreign body were treated with self-made intraocular earth magnet (IOEM): 2 patients with foreign body remained on lens, 7 on posterior segment of global wall, 12 lodged in the post pole of retina and 1 on the surface of optic disk. These patients were operated under microscope. The foreign bodies on the posterior segment were removed by using IOEM combined with pars plana vitrectomy. RESULT: The foreign bodies of these 22 patients were successfully removed with one operation respectively. The minimum volume of the foreign bodies was 0.5 x 0.5 x 0.5 mm3. The maximum volume of the foreign bodies was 5 x 2 x 1.5 mm3. The average maximum diameter of the foreign bodies was 2.47 +/- 1.2 mm. CONCLUSION: Intraocular earth magnet (IOEM) is a permanent magnetic instrument with moderate magnetic power. Direct observation and detachment of the foreign body with ocular tissue are necessary during the remowal of the forreign body. This instrument is suitable for intraocular microsurgery, can be handled easily, used for extraction of foreign bodies in anterior chamber, on turbid lens and in vitreous body, in removing foreign bodies in vitreous chamber with obvious injured retina and on posterior segment, the surface of retina, and can also be applied toextract foreign bodies on optic disk as well asforeign bodies partly oldged on global wall. PMID- 9208664 TI - [Histopathologic classification of 1921 orbital tumors]. AB - PURPOSE: The retrospective study is undertaken to determine the histopathologic types of space-occupying lesions of the orbit in 1921 cases examined between 1953 1992 at the Eye Pathology Laboratory of Shanghai Medical University. METHODS: The authors reviewed old diagnostic pathologic slides on file including HE stain as well as special stains, if available, for all the indeterminate cases. Newly prepared immunohistochemical stains are made for the controversial cases to help diagnosis. Some slides are discussed and diagnosed by Pathology Slide Meeting of the Shanghai Surgical Pathology Society. Cases with diagnosis not agreed upon by the pathology meeting are excluded from this study. RESULTS: The 5 leading malignant orbital tumors are malignant lacrimal gland tumors (138 cases, 32%), malignant lacrimal sac tumors (112 cases, 26%), rhabdomyosarcomas (65 cases, 15%), lymphosarcomas (56 cases, 13%), and malignant hemangiopericytomas (31 cases, 7%). The 5 leading benign orbital tumors are cavernous hemangionas (411 cases, 36%), dermoid cysts (152 cases, 13%), benign mixed tumors (150 cases, 13%), inflammatory pseudotumosr (129 cases, 11%), and schwannomas (92 cases, 8%). Rare tumors of the orbit include alveolar soft tissue sarcoma, chondrosarcoma, mesenchymal chondrosarcoma, synovial sarcoma, giant cell tumor, granular myoblastoma, yolk sac tumor, and retinal anlage tumor. CONCLUSION: This study, in agreement with other reports of the statics study of the orbital tumors demonstrates the prevalence of hemangiomas, lacrimal gland tumors and lymphoid pseudo-tumors to be the common space-occupying lesions in the orbit. Some rare tumors are discussed. PMID- 9208665 TI - [Vitrectomy combined with scleral buckling for the treatment of complicated retinal detachment]. AB - PURPOSE: To raise operative successful rate of complicated retinal detachment, we used the methods of vitrectomy combined with scleral buckling for the treatment of complicated retinal detachment. METHODS: The operative methods included: scleral buckling, vitrectomy, membrane peeling, intraocular and extraocular drainage, gas and silicone oil tamponade. RESULT: Being followed up for 1-16 months, 22 eyes got anatomic reattachment in 29 eyes, the operative successful rate was 75.8%; 15 eyes were injected with silicone oil intraocular tamponade, with 12 eyes getting reattachment. The successful rate was 80%. CONCLUSION: Since the vitreous surgery is used, it is possible for the treatment of complicated retinal detachment, the operative successful rate can be raised. The technique of membrane peeling and intraocular tamponade was very important. Silicone oil has been accepted by more and more ophthalologists in the world, which acts as an effective material of intraocular tamponade. PMID- 9208667 TI - [Clinical application of silica ball implant enclosed by an autologous reversal sclera in patients with enucleation]. AB - PURPOSE: To observe the effects of improved silica ball implantation enclosed by autologous reversal sclera. METHODS: After the enucleation of eye balls, we reversed, cleared the autologous sclera, enclosed the silica ball and implanted it in the operative eye. RESULTS: 30 cases with enucleation were implanted the prosthetic eyes. The follow-up period was 6 months to 3 years. The prosthetic eyes had the motility of 10 degrees to 25 degrees and the empty socket syndrome was effectively prevented. CONCLUSION: The sargical method can overcome the defect that prosthetic eyes cannot rotate after being implanted and has got satisfactory cosmetic effects. PMID- 9208666 TI - [The application of the pattern visual-evoked potential in the diagnosis and treatment of amblyopia]. AB - PURPOSE: To research into the value of the application in the diagnosis and treatment of amblyopia, and to discuss the mechanism of amblyopia. METHODS: The subjects were divided into three groups: 108 amblyopia eyes; 180 normal eyes and 26 amblyopia eyes during the treatment with Madopar. PVEP was observed with black white checks pattern-reversal stimulation at several spatial frequencies (30', 60', 90'). RESULTS: P1 latency of the amblyopia eyes prolongs at the high frequency (30' angle of view), in which the occurrence rate of abnormal PVEP is 68.5%. Through detecting PVEP during the treatment of amblyopia with Madopar, we found P1 latency shortened and amplitude increased (P < 0.05). CONCLUSION: It is better to use P1 latency to observe the visual function of patients with amslyopia with stimulation at 30' angle of view. PVEP is an important objective index in diagnosis and evaluating the curative effect of amblyopia. PMID- 9208668 TI - [Diagnosis and course (under treatment) of Parkinson disease]. AB - Diagnosis of Parkinsonism is made in two steps: 1. identification of the Parkinson syndrome, a combination of rest tremor, hypertonia, akinesia and postural disturbances; 2. then essentially on the basis of clinical observations, relation to Parkinson's disease. The main risks during the course with L-dopa treatment are, on the one hand, the appearance of akinetic changes and movement disorders, more common in the younger affected patients, and on the other hand, disorders that do not respond to L-dopa, especially postural and cognitive, that are favoured by old age. Anxiety, depression pain, autonomic disorders and insomnia increase the repercussions of the disease and complicate its management. PMID- 9208669 TI - [Mechanisms and treatments of Parkinson disease]. AB - The loss of dopaminergic melanized neurons of the substantia nigra is the main lesion of Parkinson's disease. This cell death, of unknown origin, is accompanied by the formation of free radicals and the occurrence of oxidative stress. The nigro-striatal dopaminergic hypoactivity induces a disinhibition of the subthalamo-pallidal structures responsible for a frontal cortex hypoactivity, at the origin of akinesia. Levodopa is the most effective treatment of Parkinson's disease, but exposes to motor complications that new therapeutic strategies try to limit. The neurosurgical treatment, especially deep brain stimulation, is applied to few selected patients. PMID- 9208671 TI - [Chorea and Huntington's disease]. AB - The causes of chorea are numerous and it is convenient to distinguish hereditary chorea and acquired syndromes with chorea. Huntington's disease represents the most typical disorder with chorea and the clinical diagnosis must be considered if there is chorea intellectual decline and autosomal dominant inheritance. The availability of molecular analysis for diagnostic confirmation shows that the clinical diagnosis in experienced centers is very accurate. Predictive testing involves several ethical problems, especially related to the severity of the disease and its absence of treatment. The other causes of hereditary chorea are less frequent and specific clinical contexts are evoqual of acquired choreas. PMID- 9208670 TI - [Parkinson "plus"]. AB - "Parkinson plus" is a group of sporadic degenerative disorders associating Parkinsonism, poorly sensitive to L-dopa, to other neurological syndromes. Thus, progressive supranuclear palsy includes Parkinson's disease, vertical gaze paralysis, nuchal dystonia and dementia; multisystem atrophies associate to different degrees Parkinson's disease (striatonigral degeneration), a cerebellar syndrome (olivopontocerebellar atrophy), dysautonomia (Shy-Drager syndrome), pyramidal syndrome, etc. Other diseases (corticobasal degeneration, diffuse Lewy body disease) also belong to the Parkinson "plus" group. Clinical differentiation between Parkinson "plus" and idiopathic Parkinson's disease is difficult. Their prognosis and treatment are substantially different. Certain diagnosis is based solely upon anatomical observations. PMID- 9208672 TI - [Dystonia]. AB - Dystonia can be considered either as a symptom, or as a disease. An initial classification of dystonia can be made according to the localization and the severity of the spasms or the associated movement disorders such as myoclonus. A second classification differentiates idiopathic dystonia and secondary dystonia. Personnel medical history, familial cases, neurological symptoms such as pyramidal, cerebellar, oculomotor signs are helpful clues in the diagnosis strategy. Drugs, botulinum toxin, physiotherapy are often combined symptomatic treatment regardless of the cause of dystonia. PMID- 9208673 TI - [Tics and Gilles de la Tourette disease]. AB - Clinical characteristics of tic disorders are complex and difficult to ascertain, the pathophysiology remains unknown. Their diagnosis can only be performed clinically they can be classified into two groups with opposite prognosis: transient tics of children with favorable spontaneous evolution, and Gilles de la Tourette disease, a life long disorder with disappointing therapeutic possibilities. PMID- 9208674 TI - [Tremors]. AB - In this general review of tremors, one must distinguish the parkinsonian rest tremor, which concerns relaxed muscles, from other tremors that accompany muscle activities, such as maintaining a posture or executing a movement. Among various postural and action tremors, essential tremor occupies first place, in terms of its prevalence. The diagnosis of essential tremor is based on precise criteria. Often hereditarily transmitted, essential tremor can sometimes be quite disabling. Essential tremor and parkinsonian tremor are compared on an accompanying table. Other types of tremor are reviewed according to their possible prevalences. i.e. iatrogenic tremor and dystonic tremor. Particular attention is paid to orthostatic tremor, multiple sclerosis tremor and psychogenic tremor. PMID- 9208676 TI - [Movement disorders of drug origin]. AB - The involvement of a drug must be suspected in each patient suffering from a movement disorder. Besides classical neuroleptics used as antipsychotics (butyrophenones, phenothiazines or benzamides), many drugs, mainly "hidden" neuroleptics (prescribed as antinausea, antivomiting, antivertigo, antispasmodic or antihypertensive drugs) or agents prescribed in psychiatric (antidepressants, lithium) or neurological (levodopa in Parkinson's disease, antiepileptics) diseases are known to be able to reveal or produce a movement disorder. Other drug prescribed in internal medicine can also be involved. This review discusses the main characteristics of drug-induced movement disorders as well as their pharmacological approach. PMID- 9208675 TI - [Myoclonus]. AB - Myoclonus (myos: muscles and klonos: agitation. jerk) reflect some dysfunction of the central nervous system. Their multiform clinical presentations correlate to various physiopathological mechanisms and neuropharmacological substrates, still imprecisely known. The etiologies are quite numerous but it is important to distinguish from the physiologic forms epileptic, symptomatic, essential or segmental myoclonus. The treatment, largely depending of the underlying cause, mainly includes clonazepam and valproate. PMID- 9208677 TI - [Radiotherapy. Radiobiological notions, main side effects]. PMID- 9208678 TI - [Hypereosinophilia, Diagnostic orientation]. PMID- 9208679 TI - [Herpetic encephalitis, Diagnosis, treatment]. PMID- 9208681 TI - [Acute hemodynamic pulmonary edema (cardiogenic or overload). Etiology, physiopathology, diagnosis, management in emergency situation with drug posology]. PMID- 9208680 TI - [Notions on tolerance and autoimmunity. Distinction between self and non-self. Positive and negative thymic selection; peripheric tolerance; consequences for understanding of anergy and autoimmune diseases]. PMID- 9208682 TI - [Physiopathology of closed thoracic trauma]. AB - Any thoracic trauma causes more or less severe hypoxia, due to pain, pleural effusion, mechanical ventilation disorders and pulmonary contusion. These four factors lead to bronchial congestion, which in turn aggravates hypoxia. More or less rapidly a vicious circle of respiratory failure is created. It can be broken only by early treatment of the cause. Without such treatment, respiratory failure quickly becomes autonomic and treatment of the cause is no longer sufficient. PMID- 9208683 TI - [Initial resuscitation of closed thoracic trauma]. AB - Blunt thoracic trauma are frequent and often severe. Their management is improved in the prehospital settings by the involvement of a medical team, and the admission in a specialized trauma center. Diagnostic and therapeutic steps are closely linked. They are based on the stabilization of life threatening problems and the appropriate use of recent advances in medical imaging. Endotracheal intubation and mechanical ventilation are indicated for respiratory distress. Immediate surgical management is mandatory for an hypovolemic shock related to massive hemothorax. On the contrary, multiple ribs fractures and flail chest without major pulmonary contusion are managed with regional analgesia allowing effective physiotherapy and avoiding the infectious complications related to prolonged mechanical ventilation. PMID- 9208684 TI - [Contribution of imaging in closed thoracic trauma]. AB - Conventional or digital chest radiographs are usually efficient for diagnosis of minor chest trauma. The primary modality for diagnostic evaluation of severe blunt chest trauma remains chest X-ray. Other imaging modalities especially CT have to be performed. The management of these patients is based upon clinical and initial radiographic findings. PMID- 9208685 TI - [Thoracic and abdominal wounds]. AB - Thoracic and abdominal wounds are characterized by their diversity, their possible danger and the necessity of a successful diagnosis and therapy strategy. Management of thoracic wounds and indications of surgical treatment are conditioned by airway and hemodynamic states, paraclinical exams and chest drainage. The approach of abdominal wounds is based upon their possible penetrating character. Surgical indications, even if very discussed, are still wider. Thoraco-abdominal wounds could concern the diaphragm and are remarkable for their surgical strategy. PMID- 9208686 TI - [Diaphragm rupture in closed trauma]. AB - Ruptures of the diaphram occur in approximately 2% of cases of severe thoraco abdominal trauma. They are present on the left in 80% of cases. The rupture is cupolar, sagittal or transversal, with peripheral desinsertion (the last always observed on the right), or paravertebral and retropericardiac posterior tears. Movement of the abdominal viscera toward the thorax can be progressive, with signs appearing only after 3 or 4 days or more. Any attempt to evacuate an intrathoracic effusion should be made carefully. In half the cases in polytraumatic patients, the lesion is confirmed by clinical suspicion or by appropriate surgical exploration. The first emergency step is repair. In cases of recent occurrence and for reasons of abdominal safety, coeliotomy is preferred. Mortality ranges from 20 to 30% and depends on the polytraumatic state but also on heart and respiratory failure and on infectious complications. PMID- 9208687 TI - [Closed abdominal trauma: diagnostic and therapeutic orientations]. AB - In France, traumas are the third leading cause of mortality. There are no clear statistics concerning abdominal injuries among all traumas. Nevertheless abdominal traumas are directly responsible for 10 to 30% of traumatic death. Over the past few years, improvement in imaging technics has allowed to establish very precise damage toll and to consider new therapy approaches. The development of conservative treatments has led to a reduction in the number of unnecessary coeliotomy. The treatment of such pathologies requires a multidisciplinary, effective and constantly available staff. PMID- 9208688 TI - [Imaging of abdominal trauma]. AB - Imaging technique strategies in abdominal traumas should be aimed at determining the best strategy allowing a reliable and quick diagnosis of a potential surgical lesion. The introduction and development of modern imaging methods including CT and US thus nowadays, have a clear impact on the diagnosis and treatment of abdominal traumas. PMID- 9208689 TI - [Role of surgery in closed abdominal trauma]. AB - Over the past twenty years, nonoperative management has increasingly been recommended for the care of patients with blunt abdominal trauma. Emergency laparotomy remains the rule in patients with hemodynamic instability or in those with peritonitis due to intestinal perforation. Surgical treatment of liver and splenic lesions tends to be more conservative. After assessment of the lesions by computed tomography, nonoperative management in intensive care unit is allowed in the majority of patients. PMID- 9208690 TI - [Malnutrition. Clinical and biological signs, treatment]. PMID- 9208691 TI - [Obstruction of the central artery of the retina and its branches. Etiology, diagnosis, course, treatment]. PMID- 9208692 TI - [Atopic dermatitis. Diagnosis, course, treatment]. PMID- 9208693 TI - [CMV infections in patients infected by HIV. Diagnosis, course, principles of treatment]. PMID- 9208694 TI - [Myasthenia syndrome. Diagnostic orientation]. PMID- 9208695 TI - [The cancerous cell. Genetic abnormalities (oncogenes and anti-oncogenes), growth factors, apoptosis and dissemination factors, prognostic incidence]. PMID- 9208696 TI - [Infectious mononucleosis. Epidemiology, diagnosis, course]. PMID- 9208741 TI - Saving our children from children. Conquering teen pregnancy challenges for our youngest mothers. PMID- 9208742 TI - Are nurses ignoring the call for advanced skills? PMID- 9208743 TI - Being proactive about assistive personnel. PMID- 9208744 TI - Thinking globally, acting locally. Advocating for health care policy in your community. PMID- 9208745 TI - Documenting a neonate's death. PMID- 9208746 TI - Silent suffering. Helping women find the path to continence. PMID- 9208747 TI - Pressuring pain. Alternative therapies for labor pain management. PMID- 9208748 TI - Preventing preterm labor. Is terbutaline our best option? PMID- 9208749 TI - A triggering time. Childbirth may recall sexual abuse memories. PMID- 9208750 TI - Caring for caregivers. Proactive planning eases burdens on caregivers. PMID- 9208751 TI - Placental evaluation: pregnancy's black box? PMID- 9208753 TI - Prep your patients for menopause. PMID- 9208752 TI - Marketing your expertise. Discussion groups promote self-care; your facility. PMID- 9208754 TI - The individuality of grief. PMID- 9208755 TI - Is technology stealing the "heart" of nursing? PMID- 9208756 TI - [The state and the commercialization of public health institutions]. PMID- 9208757 TI - [The regional characteristics of the health of the population and policy in the public health field]. AB - The authors analyze regional specificities in the health status of residents of St. Petersburg. They emphasize that public health strategy should be based on specific features of individual regions and the relevant methods of public health maintenance. PMID- 9208758 TI - [Relations within the family as a health factor for adolescent girls]. AB - The authors attempt to assess the effect of everyday relationships in a family on the health status of family members, adolescent girls. They emphasize the importance of further investigation and analysis of everyday family life, for the data of such studies will help plot new multifactorial models aimed at the maintenance, fortification, recovery, and development of the latent potentials of a family in order to provide its normal functioning in the society and adequate adaptation to ever changing socioeconomic conditions. PMID- 9208759 TI - [Disability as a medico-demographic problem and the mortality of the disabled]. AB - Socio-hygienic analysis of the mortality of disabled subjects is carried out on the basis of clinical and socio-statistical records with due consideration for the levels and structure of death causes, types and groups of disability, duration of stable disability, etc. Scientific approaches to analysis of mortality of disabled subjects as a medico-demographic problem have been outlined and basic information singled out to be used in development of special medicosocial programs. PMID- 9208760 TI - [Current problems and the development of public health in Russia]. PMID- 9208761 TI - [Current problems in preventive medicine]. AB - The author presents data on the incidence of diseases involving temporary invalidity among workers of the Moscow Railway. The difference in morbidity levels at various enterprises in this branch of industry is as high as 2.9 to 5.9 times, which fact is regarded as a sign of uncontrollable increase of morbidity among the workers of the predominant part of the production. A system of registration and organizational measures aimed at morbidity prevention, conditionally named Automated Monitoring System 'Morbidity' (AMSM) is presented. Results of distribution of morbidity by "days" in the final line 30 of record file 16BH in relative compatible parameters are described. Introduction of AMSM resulted in reduction of morbidity in 1993 as reflected in line 35 in "days" by 10% vs. the year 1992 and in a different rating of a test enterprise TC-18 in the total group of the Moscow Railway Depot, which moved from the 4th to 10th position. Economical estimation of the benefit due to morbidity reduction and validation of cost efficacy and self support of AMSM are presented. PMID- 9208762 TI - [A diagnostic center in the practical public health system]. AB - The authors describe a clinical diagnosis center working in close cooperation with a chair of therapy. Analysis of the activities of the center demonstrated the usefulness of medical divisions of such kind for practical health service, for it is at such diagnostic centers that profound examinations of patients, making use of present-day diagnostic methods, is possible at the preclinical stage. Such a possibility was offered to the residents of a large region who previously had to be hospitalized for this purpose. A diagnostic center brings evident economic benefit and is a clinic where physicians of primary health centers upgrade their knowledge and skills. PMID- 9208763 TI - [The principles of designing the information infrastructure in the military medical service]. PMID- 9208764 TI - [The forms and methods of specialist postgraduate training]. PMID- 9208765 TI - [The technology of introducing the system of obligatory medical insurance for the rural population]. AB - The paper deals with organization of obligatory medical insurance of the rural population of Moscow Province. Present-day public health system provides virtually unequal availability of medical care rendered to the urban (compactly living) and rural (living at larger territories than in cities) residents. The aim of the law "On Medical Insurance of the Citizens of the Russian Federation" as far as it concerns medical care rendered to any population groups in accordance with requirements in such care should be implemented by the local administration, including newly created or specialized funds of obligatory medical insurance, organs of public health management, and financial organs. The system of regulation created by these organs should provide equalization of the scope and quality of medical care rendered to the urban and rural population. PMID- 9208766 TI - [Experience in the elaboration of a program of public health development with medical insurance for an industrial region]. AB - Requirements in medical care and availability of such care for the population of an industrial regions with the towns of Staryi Oskol and Gubkin is analyzed. The authors emphasize the unique type of the region. Approaches to improvement of the level of public health in this region at the expense of coordination of the activities of medical and public health institutions, equipment of therapeutic and prophylactic institutions with modern therapeutic and diagnostic devices, and upgrading the knowledge and skills of physicians and paramedical personnel, which may be implemented by professors and teachers of the N. N. Burdenko Voronezh Medical Academy, are outlined. The possibility of creation of specialized medical centers at large multiprofile hospitals is discussed. PMID- 9208767 TI - [A mathematical approach to the establishment of criteria for assessing the quality of medical services]. AB - The paper devoted to characterization of the quality of medical services offers only vague descriptive definitions of this notion. The market of medical services necessitates development of objective and accurate criteria of the final results of work compatible with the expenditures. The present paper offers criteria for assessment of the quality of medical work in the form of coefficients of efficiency, availability, and emergency. PMID- 9208768 TI - [The history of insurance medicine in Saint Petersburg]. PMID- 9208769 TI - [The experience of cooperative work by therapists and surgeons in wartime and under extreme peacetime situations]. PMID- 9208770 TI - [The classification of the sources in medical history research]. PMID- 9208771 TI - [A prominent Russian hygienist (on the 150th anniversary of the birth of V. A. Subbotin)]. PMID- 9208772 TI - [The development of social hygiene science and the contribution of the N. A, Semashko Research Institute of Social Hygiene, Economics and Public Health Management in this field of medicine]. PMID- 9208773 TI - [Health and the sociomedical needs of the young family]. AB - Results of a comprehensive socio-hygienic examination of a young family with a baby aged under one are analyzed. The authors disclose the notion of the reproductive function and health of a woman in connection with childbirth, upbringing of children, and meeting the basic requirements of an infant under one in medical care and in creation of an environment needed by such an infant. The authors present an assessment of the readiness of a young family to childbirth and upbringing of children, analyze the role of a mother in the formation of the health status of her children and of a family as a whole, and classify the families by their readiness to childbirth and to upbringing of children a relationship between the behaviour of a young mother and her child's health and quality of care of a child in a family is analyzed. The authors come to a conclusion that medicosocial requirements of a young family are governed by familial structure, health of af couple, baby's health, behaviour of a young family, and the communal conditions. PMID- 9208774 TI - [The sociomedical problems of occupational stress]. AB - The paper analyzes medicosocial problems of occupational stress, characteristics of stressor reactions associated with the health status, job parameters, and lifestyle of different social groups of population. The incidence of stress among various occupational groups in the system of the Ministry of Internal Affairs of Kazakhstan has been studied. The authors have detected the role of external and internal environmental factors (social, demographic, biological) in the appearance and development of stress. Recommendations are offered on the detection of stress, identification of groups by the severity of stressor reactions, and formation of risk groups. The scope and type of therapeutic and prophylactic care in specific groups followed up is outlined. PMID- 9208775 TI - [Seasonality and population mortality]. AB - The authors analyze clinical records and discuss the problem of a relationship between population mortality rates and seasons. They point to a reliable correlation between cyclic changes in weather and climate and death rates due to a number of causes. The importance of bearing in mind the detected regularities when planning and utilizing the resources of specialized medical care to prevent and arrest the critical states developing at certain periods is emphasized. PMID- 9208776 TI - [The conceptual trends in the development of a model of primary health care in Ukraine under the current conditions]. AB - The authors validate the necessity of reformation of the system of primary health care in the Ukraine. They analyse the solution of this problem in various countries of the world and outline the major trends in development of the model of primary health care in the Ukraine under present-day conditions. PMID- 9208777 TI - [Territorial public health services and their management under the new economic conditions]. PMID- 9208778 TI - [Improvement in the sociomedical support for the urban family under current conditions]. AB - Medico-demographic processes and socio-hygienic characteristics of the population of a large city, both at an individual and familial levels, as well as territorial characteristics of residence, should be taken into consideration when developing measures for medicosocial care of the population. Analysis of the attitude of 380 city families to the activities, personality, and professional skills of local therapists and pediatricians and to the idea of primary health care rendered by a physician of a general profile demonstrated an increased activity of the families in the "physician-patient" system and confirmed the necessity of improving medicosocial care of a city family, expansion of prophylactic work with the population with due consideration for factors which have an impact on population health. PMID- 9208779 TI - [Methodological bases for monitoring the health of the population]. AB - Monitoring is widely used in public health nowadays. A vast scope of data has been accumulated in the latest decade on the regularities of population health formation. Risk factors and the degrees of their unfavorable effects on the population health have been classified. This created conditions for the development of information systems for not only monitoring, but for active intervention in the processes of health formation. With the same purposes in view a system of cooperation with various population groups is created based on their informed consent. The paper discusses the notions and definitions in this field, methodologic basis provision of the information needed for monitoring, suggests criteria for selection of factors to be followed up and the organisation and methodological scheme of experimental validation of socio-hygienic monitoring of the population health. PMID- 9208780 TI - [Monitoring in the system of specialized medical care management (exemplified by the study of burns--specialized care for burn victims)]. AB - Technological revolution involves an increased risk of negative impact of adverse factors on human health in the whole world. The authors analyze the tendencies of burn traumas incidence in Russia in the latest decade. They necessitate creation in each district of centers for monitoring burnt patients, which should be engaged in registration of burn traumas, organization of emergency care and specialized outpatient and inpatient care, medical and sociopsychological rehabilitation of the victims. PMID- 9208781 TI - [An automated work station for the physician of a city polyclinic]. PMID- 9208782 TI - [Public health reforms in the countries of central and eastern Europe--how they see them from western Europe]. PMID- 9208783 TI - [The process of public health reform in Sweden]. PMID- 9208784 TI - [Reform of the financing of the public health system in Israel]. PMID- 9208785 TI - [Public health in the upper Volga valley during World War II]. PMID- 9208786 TI - [An innovator in military medicine (on the centenary of the birth of N. I. Zavalishin)]. PMID- 9208788 TI - [Health and the sociomedical problems of the family]. PMID- 9208789 TI - [The state and dynamics of disability in childhood in the Russian Federation]. AB - Characterizes the state and time course of childhood disability in Russia as reflected in statistical files of the service of social protection of the population. Presents the invalidity level in childhood: 79.7 per 10,000 population aged under 16 in 1992. The level of childhood invalidity increased by 4.8 times over 13 years. An unfavorable prediction regarding invalidity in 2000 is given. Epidemiological assessment of childhood invalidity as a regional level and for some territories is presented. Territories with stable and unstable levels of childhood invalidity were detected. Classification of territories of the Russian Federation by levels and rates of growth of childhood invalidity was made. Recommendations to solve present day problems of childhood invalidity are offered. PMID- 9208787 TI - [Physicians of Moscow University in the pages of the newspaper Russkie Vedomosti]. PMID- 9208791 TI - [Premature mortality in the working-age population of Russia]. AB - Basing on analysis of statistical data, the author examines the level, time course, and structure of mortality of the able to work population of Russia. The notion of premature mortality, its level, social and medical causes are discussed. The significance of inter-branch integration and of complex efforts to reduce the unjustified loss of population is emphasized. PMID- 9208790 TI - [The scientific rationale and basic methodological principles for creating government statistics on childhood disability in the Russian Federation]. AB - The authors necessitate the creation in Russia of a new system of state statistical registration of childhood invalidity based on territorial registers of disabled children, which will make available the statistical data characterizing this population both at the regional and federal levels of government. At present such information in our country is virtually unavailable, this impeding the development of scientifically based programs of medicosocial and pedagogical aid to such children and their families. Methodological principles of organization of such a register at a territorial level are described, which will permit including the information collection for the register into the already existing in Russia system of official registration of disabled children and coordinate it with the international recommendations on relevant problems. PMID- 9208792 TI - [A medico-demographic and epidemiological analysis of digestive organ pathology in the Udmurt Republic]. AB - The incidence and causes of gastroenterological diseases in the population of Udmurtia are analyzed. Factors of risk of such diseases were defined and the "portraits" of patients with gastric and hepatic diseases designed. The quality of medical care rendered to the said patient population was assessed. The results of the study helped develop a republican program aimed at improvement of gastroenterological service and at prevention of diseases of the organs of digestion. PMID- 9208793 TI - [Medical insurance for working people and government health inspection]. PMID- 9208794 TI - [Actuarial calculations in medical insurance and the problems in their data processing support]. PMID- 9208795 TI - [The development of primary medical health care in Russia]. AB - The socio-ethical aspect in the development of primary health care in Russia is discussed. The authors analyze the role of this type of care in provision of available and free of charge, which is important in principle, medical care at the level of the primary contact between a patient and medical worker. Primary health care under present-day conditions should occupy one of the principal positions in the public health reforms. The major approaches to priority development of health care are considered, including the renaissance of the progressive traditions of zemstvo medicine, development of new approaches to redistribution of scarce resources of public health on the basis of social justice principle, social control of the distribution of resources, purposeful attention to personnel training, including training of general practitioners (family physicians), solution of a number of organizational, medicosocial, legal, ethical, and other problems, and expansion of contacts with popular medicine. PMID- 9208796 TI - [The reorganization of the administrative and management apparatus of public health with the introduction of medical insurance (based on data from Reutovo, Moscow Province)]. AB - Implementation of the Law "On Medical Insurance of Citizens in the Russian Federation" and of the relevant resolution of the Head of the Administration of the Moscow Province No. 31/1 of February 21, 1994 implies the reorganization of local schemes of public health management and changes in the administrative structures of public health institutions which became autonomic subjects in the period of transition to medical insurance. The author discusses two trends of the possible changes, one of which, after appropriate studies, was introduced in the system of local public health management in the town of Reutovo in Moscow Province. Rearrangement of the scheme of public health management in the town of Reutovo helped maintain and preserve the stable status of each public health institution, carry out ascertaining and licensing, and improve the quality of medical care rendered to the population, with the share of expenditures for administrative and managerial personnel reduced. PMID- 9208797 TI - [Euthanasia as a moral problem in medical activities]. AB - The problems related to physician's attitude to life and death in the framework of the antique and present-day medical tradition are discussed. The maintenance of the historical deontological tradition in present-day society is possible via cultivation of the art of intercourse between the physician and incurable patient and development of the psycho-therapeutical fundamentals of medical activity. PMID- 9208798 TI - [The sociomedical support of the population in Russia (sources, history and the present day)]. PMID- 9208799 TI - [A modern historical-medical analysis of the creative legacy of N. I. Pirogov]. PMID- 9208800 TI - [E. E. Fromgol'd and Moscow therapy in the'20s and 30's]. PMID- 9208802 TI - [The role of the Medical Department of Moscow University in the development of hematology in Russia]. PMID- 9208801 TI - [The sociomedical problems in the opening up of the Northern Territory in the first half of the 30's]. PMID- 9208803 TI - [Russian sisters of charity in the Crimean Campaign of 1854-1856 (on the centenary of the death of Ekaterina Mikhailovna Bakunina--1812-1894)]. PMID- 9208804 TI - [Population health in programs of medical ecological screening]. AB - Socio-hygienic analysis of population health in Ryazan Province, specifically, in the Pronya River basin, revealed a number of local features: reduced birth rate, aging of the population, high incidence of acute enteric and other infections, tuberculosis, malignant tumors, diseases of the blood and hemopoietic organs in children, frequent complications of pregnancy and labor in children which are explained to a great measure by socioeconomic and socio-hygienic condition, as well as the status of the environment, of water among other things. Correlation coefficients were derived reflecting relationships between morbidity levels and the quality of potable water and foodstuffs, availability of centralized water supply and sewer system, as well as atmospheric air pollution. The data help better validate concrete measures in the program of sanitization of the population in these regions. PMID- 9208805 TI - Human papillomavirus, integration and cervical carcinogenesis: a clinicopathological perspective. PMID- 9208806 TI - Kaposi's sarcoma associated herpes virus (KSHV/HHV 8): epidemiology, molecular biology and tissue distribution. PMID- 9208807 TI - Intracellular staining of Mx proteins in cells from peripheral blood, bone marrow and skin. AB - AIM/BACKGROUND: The Mx proteins are known to be specifically and dose dependently induced in mononuclear cells (MNC) by type I interferons (IFN). The aim of this study was to establish a staining method for the human intracellular Mx proteins, MxA and MxB, in leucocytes and bone marrow and skin cells. METHODS: Several monoclonal antibodies directed against the MxA and MxB proteins were generated. These antibodies were used to stain Mx proteins in both frozen and paraffin wax sections using the standard alkaline phosphatase anti-alkaline phosphatase (APAAP) method. RESULTS: Granulocytes, monocytes and lymphocytes extracted from freshly collected blood from 21 healthy subjects did not stain. After incubating MNC from these subjects with IFN alpha 2b for 48 hours, Mx proteins were detected in monocytes and lymphocytes. Within two days of starting treatment with subcutaneous IFN alpha 2b, granulocytes, monocytes and lymphocytes of 16 patients with cancer stained strongly for Mx proteins. The intensity of staining was correlated with the Mx content of whole blood measured using a specific ELISA. Prior to IFN treatment, cells from bone marrow and skin tissue specimens were negative for Mx proteins with the exception of endothelial cells. During treatment with IFN alpha 2b, nearly all cells from bone marrow and skin stained intensely. CONCLUSIONS: These new monoclonal antibodies facilitate the detection of Mx positive cells in peripheral blood and in frozen or paraffin wax specimens. The advantage of this staining method is that individual cells which have responded to viruses or biologically active IFN alpha, beta or omega can be identified. PMID- 9208808 TI - Oligoclonal populations of T and B cells in a case of angioimmunoblastic T-cell lymphoma predominantly infiltrated by T cells of the VB5.1 family. AB - AIMS: Immunohistological and molecular characterisation of a case of peripheral T cell lymphoma (PTCL) of angioimmunoblastic T-cell lymphoma (AILD) type. METHODS: Frozen and paraffin wax sections of the diagnostic lymph node were stained with a panel of T- and B-cell lineage monoclonal antibodies. DNA was isolated from the paraffin wax embedded biopsy material for T-cell receptor (TCR) and immunoglobulin (Ig) PCR amplification, and resultant PCR products were cloned and sequenced. RESULTS: Immunohistological analysis of the presenting lymph node was consistent with an extensive infiltrate of pleomorphic CD3+CD8+ lymphocytes. Most (>80%) of these infiltrating CD3+ cells were also positive for the TCR VB5.1 gene family product, and were shown to be oligoclonal by TCRB PCR amplification and sequencing. Three oligoclones of B cells were also demonstrable by PCR amplification with Ig heavy chain primers and sequencing, a finding at variance with the diagnosis of AILD. CONCLUSIONS: These data demonstrate the complexity and heterogeneity of PTCL which require extensive histological examination and molecular characterisation. PMID- 9208809 TI - p53 gene mutations in multiple myeloma. AB - AIM: To assess whether p53 gene mutation is important in the pathogenesis and progression of multiple myeloma. METHODS: Thirty eight DNA samples (derived predominantly from bone marrow) obtained from 31 patients with multiple myeloma were examined for mutations in p53 exons 5-9 by polymerase chain reaction single strand conformation polymorphism. Twenty three samples were analysed at the time of diagnosis (one patient had plasma cell leukaemia), three in plateau phase, and 12 at relapse (one plasma cell leukaemia and one extramedullary relapse). RESULTS: One p53 mutation was detected in this group of patients (3.2%). This was seen in the diagnostic bone marrow sample of a 35 year old man with stage IIA disease and occurred in exon 6 as a result of a silent A to G transition at codon 213 (CGA-->CGG), a polymorphism that has been reported in about 3% of breast and lung tumours. CONCLUSIONS: p53 gene mutations are rare events in multiple myeloma and would seem to be of limited value as a prognostic factor. PMID- 9208810 TI - Immunocytochemical expression of growth factors by odontogenic jaw cysts. AB - AIM: To determine the immunocytochemical pattern of expression of transforming growth factor (TGF) alpha, epidermal growth factor (EGF), and TGF beta in the three most common types of odontogenic jaw cyst. METHODS: Growth factor expression was detected in paraffin wax sections of odontogenic cysts (27 odontogenic keratocysts, 10 dentigerous cysts, and 10 radicular cysts) using a streptavidin-biotin peroxidase technique with monoclonal antibodies directed against TGF alpha (clone 213-4.4) and TGF beta (clone TB21) and a polyclonal antibody directed against EGF (Z-12). RESULTS: The epithelial linings of all cysts showed reactivity for TGF alpha which was mainly localised to basal and suprabasal layers. Odontogenic keratocyst linings expressed higher levels of TGF alpha than those of dentigerous and radicular cysts, with 89% (24/27) of odontogenic keratocysts exhibiting a strong positive reaction compared with 50% (five of 10) of dentigerous and radicular cysts, respectively. EGF reactivity was similar in all cyst groups, weaker than that for TGF alpha and predominantly suprabasal. TGF alpha and EGF were also detected in endothelial cells, fibroblasts and inflammatory cells within the cyst walls. The most intense TGF beta staining in odontogenic cysts was extracellular within the fibrous tissue capsules, irrespective of cyst type. CONCLUSIONS: These results, together with previous studies of EGF receptor, indicate differential expression of TGF alpha, EGF and their common receptor between the different types of odontogenic cyst, suggesting that these growth factors (via autocrine or paracrine, or both, pathways) may be involved in their pathogenesis. PMID- 9208811 TI - Expression of bcl-2 in bladder neoplasms is a cell lineage associated and p53 independent event. AB - AIMS: To investigate bcl-2 and p53 protein expression in hyperplastic, metaplastic and neoplastic epithelia of the urinary bladder in relation to cell lineages (transitional versus glandular epithelia). METHODS: Formalin fixed, paraffin wax embedded archival tissue blocks of 29 transitional cell carcinomas (TCC), 11 adenocarcinomas, five specimens of cystitis glandularis, four papillomas, and seven samples of morphologically normal bladder mucosa were examined immunohistochemically with antibodies specific to bcl-2 and p53. Consecutive sections were used to assess co-expression of the two proteins. RESULTS: bcl-2 protein was expressed heterogeneously in basal cells of the normal transitional epithelium, whereas p53 was rarely detectable in either normal or hyperplastic epithelium. Of the 29 TCCs, 20 (69%) expressed immunodetectable p53 which was positively associated with grade. In contrast, bcl-2 was detected in four (14%) TCCs and its expression was not associated with grade. bcl-2 was expressed constitutively in all five specimens of cystitis glandularis and in all adenocarcinomas; p53 was co-expressed in most of the latter. There was no association between bcl-2 and p53 protein expression in the TCCs. Expression of bcl-2 protein correlated negatively with grade of adenocarcinoma. CONCLUSION: In bladder adenocarcinomas, bcl-2 expression correlated negatively with tumour grade whereas p53 was associated positively with tumour grade. The association of bcl-2 with cystitis glandularis and adenocarcinoma but not TCC suggests that it may be involved in triggering a lineage switch converting transitional epithelium to a glandular phenotype. PMID- 9208813 TI - Determination of penicillin susceptibility of Streptococcus pneumoniae using the polymerase chain reaction. AB - AIM: To develop a polymerase chain reaction (PCR) based method to detect penicillin susceptibility in isolates of Streptococcus pneumoniae (SP). METHOD: PCR primers were designed to amplify differential nucleotide sequences of the penicillin-binding protein (PBP) genes 2b, 2x, and 1a in penicillin susceptible and resistant strains of SP. Primers derived from the PBP 2x and 2b genes were designed to amplify products from penicillin susceptible S pneumoniae (PSSP), whereas primers derived from the PBP 1a gene were designed to amplify gene sequences of penicillin resistant S pneumoniae (PRSP). RESULTS: Two hundred and thirty clinical isolates of SP from the USA, UK, Kenya, Romania, and the Kingdom of Saudi Arabia were tested. Of the isolates, 116 were penicillin susceptible, 65 were intermediately resistant, and 49 were highly resistant. PCR identified 108 (93%) of 116 of PSSP isolates, 55 (85%) of 65 intermediately resistant isolates, and all of the 49 highly resistant isolates of SP. The susceptibility of 16 (7%) isolates could not be determined using PCR. All of these 16 isolates had a minimum inhibitory concentration (MIC) of penicillin < 1 mg/l. None of the highly resistant isolates was identified as penicillin susceptible by PCR, although two of the isolates with intermediate resistance (MIC = 0.125 mg/l) were. CONCLUSION: Using primers that differentially identify the genotypes of susceptible and resistant strains of SP, PCR provides a rapid method for determining the penicillin susceptibility of SP isolates and could potentially be used for testing clinical samples. PMID- 9208814 TI - Male pseudohermaphroditism resulting from a novel mutation in the human steroid 5 alpha-reductase type 2 gene (SRD5A2). AB - The enzyme steroid 5 alpha-reductase, via NADPH, catalyses the conversion of testosterone to dihydrotestosterone, which is required for the embryonic differentiation of the external male genitalia and the prostate. An impairment of this reaction causes a form of male pseudohermaphroditism in which genetic males differentiate predominantly as phenotypic females. Molecular analysis of the 5 alpha-reductase type 2 gene in a patient with confirmed biochemical 5 alpha reductase deficiency has resulted in the identification of a novel mutation, GAA to AAA, at codon 200. This mutation produces an amino acid change from glutamic acid to lysine, and may affect the ability of the enzyme to bind its co-factor. PMID- 9208812 TI - The product of the imprinted H19 gene is an oncofetal RNA. AB - AIMS/BACKGROUND: The H19 gene is an imprinted, maternally expressed gene in humans. It is tightly linked and coregulated with the imprinted, paternally expressed gene of insulin-like growth factor 2. The H19 gene product is not translated into protein and functions as an RNA molecule. Although its role has been investigated for more than a decade, its biological function is still not understood fully. H19 is abundantly expressed in many tissues from early stages of embryogenesis through fetal life, and is down regulated postnatally. It is also expressed in certain childhood and adult tumours. This study was designed to screen the expression of H19 in human cancer and its relation to the expression of H19 in the fetus. METHODS: Using in situ hybridisation with a [35S] labelled probe, H19 mRNA was detected in paraffin wax sections of fetal tissues from the first and second trimesters of pregnancy and of a large array of human adult and childhood tumours arising from these tissues. RESULTS: The H19 gene is expressed in tumours arising from tissues which express this gene in fetal life. Its expression in the fetus and in cancer is closely linked with tissue differentiation. CONCLUSIONS: Based on these and previous data, H19 is neither a tumour suppressor gene nor an oncogene. Its product is an oncofetal RNA. The potential use of this RNA as a tumour marker should be evaluated. PMID- 9208816 TI - Monitoring PREP: an exercise in trust and accountability. PMID- 9208815 TI - Additional TCRV beta primers and minor method modifications improve detection of clonal T-cell populations by RT-PCR. AB - The TCRV beta RT-PCR method for detection of clonal populations of T cells which we described previously could not detect clones that used certain variable (V) beta region families. V beta 2, 4, 8.3, and 18 had insufficient homology with the original consensus V region primer. Two new primers have been designed which work well and are able to amplify from V beta families previously undetectable by this RT-PCR. In addition, minor alterations to the cDNA synthesis and gel analysis of the PCR products make the results even easier to interpret. All the Diversity/Joining (D/J) region primer combinations except for D2/J2 have been omitted, and terminating the reverse transcription by heating prior to PCR greatly improves amplification with these primers. Use of 8% and/or 10% polyacrylamide gels increases clarity. Inclusion of the modifications described will reduce false reporting of patients as having a polyclonal T-cell population. PMID- 9208817 TI - Writing a letter of application. PMID- 9208818 TI - Chronic obstructive pulmonary disease. PMID- 9208819 TI - Healthcare evaluation: evaluating the health service. PMID- 9208820 TI - Effective note-taking. PMID- 9208822 TI - NLN conference explores the future of gerontological nursing. PMID- 9208821 TI - National vocational qualifications? PMID- 9208824 TI - Enrollments in basic RN programs declining. PMID- 9208823 TI - Nurse practitioner educators join effort to improve access to primary care New York City. PMID- 9208825 TI - Evaluation of pelvic fractures. Clinical and radiologic. AB - This article gives the orthopedic surgeon a framework to use during the initial evaluation of a patient with a pelvic fracture in the emergency room. The essential elements of the assessment of instability, both clinically and radiologically, are given in this article. Examples of the major patterns of instability are provided. PMID- 9208826 TI - Assessment and management of pelvic fracture in the hemodynamically unstable patient. AB - Hypotensive patients with pelvic ring injuries present a diagnostic and therapeutic challenge. This article reviews pelvic anatomy, the classification of pelvic injuries, and how to rapidly identify patients' unstable pelvic injuries. Current recommendations for the evaluation and treatment of these patients are reviewed. PMID- 9208827 TI - External fixation for pelvic ring disruptions. AB - The application of external fixation for acute treatment of unstable pelvic fractures can be a lifesaving procedure; however, it must be coordinated with other efforts of the trauma team. The patient with a pelvic fracture must be adequately resuscitated and carefully evaluated. This evaluation includes a careful physical examination and radiographic studies, which include plain films and computerized tomography. A proper evaluation enables classification of the pelvic injury and appropriate selection of patients that require acute pelvic external fixation. In this article, both open and percutaneous techniques for pin placement and fixator frame configurations are discussed. PMID- 9208828 TI - Open pelvic fractures. A multicenter retrospective analysis. AB - A marked discrepancy exists in the reported mortality rates in patients with open pelvic fractures, ranging from 4.8% to 50%. A retrospective review of patients with open pelvic fractures was performed at three centers. Thirty-nine patients with open pelvic fractures were identified; the average age was 32. The average injury-severity score was 29 (13-75). There were 10 (26%) deaths. Factors that correlated with mortality and morbidity were instability of the pelvic fracture and the presence of a rectal injury. Delay in performing diverting colostomy correlated with a poor outcome. Previously described methods of treatment are still valid; however, there is a need for re-emphasis of early diverting colostomy in the patient with a rectal or perineal injury. A classification system for open pelvic fractures is proposed in this article. PMID- 9208829 TI - Biomechanics of pelvic fixation. AB - This article reviews a series of biomechanical studies performed in the author's laboratory, evaluating the instabilities produced by the more common and problematic pelvic ring fracture patterns and the increasingly popular and newly developed modes of internal fixation. Topics that are discussed include the following: anteroposterior compression injuries and the disrupted sacroiliac joint; sacral fractures; anteroposterior compression injuries and symphyseal fixation; new symphyseal plate designs; unstable pubic rami fractures; unstable fractures of the iliac wing; and resuscitation fixator biomechanics. PMID- 9208830 TI - Stabilization of pelvic ring disruptions. AB - Pelvic ring disruptions are challenging management problems for the orthopedic surgeon. Early hemorrhage, permanent nerve injury, and late pain caused by residual pelvic deformity are some of the many complicating factors. A variety of treatment alternatives are available to stabilize the disrupted pelvic ring. Each technique has inherent advantages and problems. PMID- 9208831 TI - Management of pelvic fractures with concomitant urologic injuries. AB - Controversy exists regarding the management of pelvic fractures with concomitant injuries to the lower urologic tract. This can be seen in both the urologic and orthopedic literature as it pertains to the ideal form of treatment for either problem and the timing of surgical intervention, if it is needed. This article reviews the anatomy, mechanism of injury, diagnostic approach, and treatment. PMID- 9208832 TI - Practical management of venous thromboembolism following pelvic fractures. AB - The management of thromboembolic complications remains one of the most controversial issues in the care of patients with pelvic and acetabular fractures. Recent studies have indicated that the incidence of proximal deep vein thrombosis is much higher than was previously believed. These patients should be managed with a formal institutional protocol that includes universal prophylaxis, supplemented in some cases by screening for deep vein thrombosis. PMID- 9208833 TI - Acetabular fractures. AB - There are many questions that remain to be answered regarding the evaluation, classification, and treatment of acetabulum fractures. Without a uniform classification and method of evaluation, a review of the literature can be misleading. Acetabular fractures represent an injury to the articular surface of a major weight-bearing joint and should be treated in accordance with the same criteria used for other intra-articular fractures. PMID- 9208834 TI - Surgical approaches to the acetabulum. AB - This article presents a concise outline of the indications, surgical techniques, and approaches to the acetabulum: Kocher-Langenbeck approach, ilioinguinal approach, iliofemoral approach, combined approaches, extended iliofemoral approach, and the triradiate approach. No one surgical appproach is ideal for all fractures of the acetabulum. The type of approach used is determined by the fracture pattern and condition of the soft tissues in the area under consideration. PMID- 9208835 TI - Total hip replacement after acetabular fracture. AB - The controversies surrounding total hip arthroplasty after acetabular fracture are presented in this article. Hip arthroplasty for acute treatment of acetabular fractures is rarely indicated. In general, total hip arthroplasty should be reserved for the late salvage of hips in which symptomatic, post-traumatic arthritis has developed after acetabular fracture. Preoperative, intraoperative, and postoperative management are discussed. Modern surgical techniques may improve the long-term survival of total hip arthroplasty after acetabular fracture, particularly the acetabular component PMID- 9208836 TI - Femoral neck and ipsilateral neck and shaft fractures in the young adult. AB - Femoral neck and ipsilateral neck and shaft fractures in the young adult represent a significant source of morbidity and mortality. This article reviews the anatomy, pathophysiology, radiographic evaluation, timing of surgery, and complications in an attempt to increase recognition of these injuries and provide better patient care. PMID- 9208837 TI - Management of osteonecrosis of the femoral head. AB - Osteonecrosis of the femoral head continues to pose a therapeutic challenge to orthopedic surgeons. This pathologic process results from the death of living components of bone from mechanical and biologic factors. Diagnosis, clinical symptoms, and classification systems are discussed. Several treatment regimens and their controversies are explored in this article. PMID- 9208838 TI - Signal transduction during natural killer cell activation: inside the mind of a killer. PMID- 9208839 TI - A spontaneously arising mutation in the DLAARN motif of murine ZAP-70 abrogates kinase activity and arrests thymocyte development. AB - Development of immature CD4+ CD8+ thymocytes into functionally mature CD4+ and CD8+ T cells is driven by selection events that require signals transduced through the T cell antigen receptor (TCR). Transduction of TCR signals in the thymus involves tyrosine phosphorylation of the protein tyrosine kinase ZAP-70 by p56(lck) and results in induction of ZAP-70 enzymatic activity. We have identified a novel, spontaneously arising point mutation within a highly conserved motif (DLAARN) in the kinase domain of murine ZAP-70 that uncouples tyrosine phosphorylation of ZAP-70 from induction of ZAP-70 kinase activity. Mice homozygous for this mutation are devoid of mature T cells because thymocyte development is arrested at the CD4+ CD8+ stage of differentiation. The developmental arrest is due to the inability of CD4+ CD8+ thymocytes to propagate TCR signals in the absence of ZAP-70 kinase activity despite tyrosine phosphorylation of TCR-associated ZAP-70 molecules. PMID- 9208840 TI - Multistage regulation of Th1-type immune responses by the transcription factor IRF-1. AB - Eradication of a given pathogen is dependent on the selective differentiation of T helper (Th) cells into Th1 or Th2 types. We show here that T cells from mice lacking the transcription factor IRF-1 fail to mount Th1 responses and instead exclusively undergo Th2 differentiation in vitro. Compromised Th1 differentiation is found to be associated with defects in multiple cell types, namely impaired production of interleukin-12 by macrophages, hyporesponsiveness of CD4+ T cells to interleukin-12, and defective development of natural killer cells. These results indicate the involvement of IRF-1 in multiple stages of the Th1 limb of the immune response. PMID- 9208841 TI - Interferon regulatory factor-1 is required for a T helper 1 immune response in vivo. AB - The transcription factor interferon regulatory factor-1 (IRF-1) mediates the effects of IFN. No information exists on its role in lymphokine production. Protection against the intracellular pathogen Leishmania major depends on a Th1 response. Here, we show that CD4+ T cells from Leishmania-infected mice lacking one (+/-) or both (-/-) alleles of the IRF-1 gene developed a profound, gene dose dependent decrease in IFNgamma production. IRF-1(-/-) mice showed dramatically exacerbated Leishmaniasis. They produced increased Leishmania-specific IgG1 and IgE, and their CD4+ T cells produced increased IL-4, characteristics of the non protective Th2 response. In cell transfer experiments, IRF-1(-/-) CD4+ T cells mounted normal Th1 responses. However, the ability of IRF-1(-/-) mice to produce IL-12 was severely compromised. Thus, IRF-1 is a determining factor for Th1 responses. PMID- 9208842 TI - Functionally distinct isoforms of STAT5 are generated by protein processing. AB - The interleukin-3 family of cytokines, which play an important role in the development of myeloid lineages, transduce signals through the JAK-STAT pathway. Previous studies demonstrate that this process entails the activation of four distinct isoforms of STAT5, where two shorter isoforms are activated in a distinct population of cells. We now demonstrate that the shorter isoforms represent carboxy-terminal truncations. Moreover, these truncations are not generated by RNA processing, but by a specific proteolytic activity. Consistent with the notion that truncated STAT5 isoforms transduce distinct signals, they fail to promote the activation of several known interleukin-3 target genes. These studies suggest that the activity of a specific protease may play a critical role in defining the biological responses transduced by STAT5. PMID- 9208844 TI - Localization, quantitation, and in situ detection of specific peptide-MHC class I complexes using a monoclonal antibody. AB - CD8+ T lymphocytes recognize antigens as short peptides bound to MHC class I molecules. Available methods cannot determine the number and distribution of these ligands on individual cells or detect antigen-presenting cells in tissues. Here we describe a method for eliciting and identifying monoclonal antibodies specific for a particular peptide-MHC class I combination. One such antibody can identify antigen complexes with a limit of detection approaching that of T cells. We used this antibody to determine the number of peptide-class I complexes generated upon viral infection, to identify antigen-presenting cells in cell mixtures, to determine the site of peptide-MHC class I interaction inside cells, and to visualize cells bearing specific peptide-MHC class I complexes after in vivo infection. Similar antibodies may prove useful for diagnostic or therapeutic purposes in cancer, infectious diseases, and autoimmune disorders. PMID- 9208843 TI - Restoration of thymopoiesis in pT alpha-/- mice by anti-CD3epsilon antibody treatment or with transgenes encoding activated Lck or tailless pT alpha. AB - Mice deficient for the pre-TCR alpha (pT alpha) chain cannot form a pre-T cell receptor (TCR) and exhibit a severe defect in early T cell development, characterized by lack of "beta selection" and impaired generation of double positive (DP) thymocytes. Here, we demonstrate that intraperitoneal injection of CD3epsilon-specific antibodies into pT alpha-/- x RAG-/- mice or introduction of an activated p56(lck) transgene in pT alpha-/- mice fully restores the number of DP thymocytes, and that expression of a transgenic pT alpha chain lacking its cytoplasmic portion can overcome all developmental defects associated with pT alpha deficiency. These results allow a better definition of the role of pT alpha in pre-TCR signal transduction and provide conclusive evidence that the cytoplasmic tail of pT alpha is not essential for pre-TCR signaling. PMID- 9208845 TI - Characterization and quantitation of peptide-MHC complexes produced from hen egg lysozyme using a monoclonal antibody. AB - Here we describe generation of Aw3.18, a monoclonal antibody that recognizes peptide residues 48-62 of hen egg lysozyme (HEL) bound to the MHC class II molecule I-Ak. Epitope mapping revealed that Aw3.18 detects a change in the solvent-exposed surface of this peptide-MHC complex upon substitution of the peptide side chain at position P1. Furthermore, Aw3.18 blocked recognition by some, but not all, of the HEL 48-62-reactive T cell hybridomas tested, suggesting a heterogeneity in the T cell response toward this complex. Finally, using Aw3.18, it was possible to determine the fraction of I-Ak molecules loaded with 48-62 peptide after culture of an antigen-presenting cell in medium containing HEL. PMID- 9208846 TI - Apoptosis signaling pathway in T cells is composed of ICE/Ced-3 family proteases and MAP kinase kinase 6b. AB - Fas/APO-1(CD95) ligation activates programmed cell death, a cellular process that plays an important role in the maturation of the host immune response. We show that activation of a specific MAP kinase kinase (MKK), MKK6b, is necessary and sufficient for Fas-induced apoptosis of Jurkat T cells. MKK6b activation occurs downstream of an interleukin-1 converting enzyme-like (ICE-like) protease(s), while execution of the apoptotic pathway by MKK6b requires both ICE- and CPP32 like proteases. Surprisingly, the p38 MAP kinase protein, a known substrate of MKK6b, does not participate in Fas/MKK6b-mediated apoptosis. These findings indicate a divergence of the MKK6b signaling pathways, one of which activates p38 and leads to regulation of gene expression, and one of which activates the ICE/Ced-3 family of proteases and leads to cell death. These studies represent a demonstration of an apoptotic pathway that is comprised of both the ICE/Ced-3 family of proteases and MAP kinase kinase 6. PMID- 9208847 TI - Casper is a FADD- and caspase-related inducer of apoptosis. AB - Caspases are cysteine proteases that play a central role in apoptosis. Caspase-8 may be the first enzyme of the proteolytic cascade activated by the Fas ligand and tumor necrosis factor (TNF). Caspase-8 is recruited to Fas and TNF receptor-1 (TNF-R1) through interaction of its prodomain with the death effector domain (DED) of the receptor-associating FADD. Here we describe a novel 55 kDa protein, Casper, that has sequence similarity to caspase-8 throughout its length. However, Casper is not a caspase since it lacks several conserved amino acids found in all caspases. Casper interacts with FADD, caspase-8, caspase-3, TRAF1, and TRAF2 through distinct domains. When overexpressed in mammalian cells, Casper potently induces apoptosis. A C-terminal deletion mutant of Casper inhibits TNF- and Fas induced cell death, suggesting that Casper is involved in these apoptotic pathways. PMID- 9208848 TI - Failure of lymphopoiesis after adoptive transfer of NF-kappaB-deficient fetal liver cells. AB - Mice deficient in the p65 subunit of NF-kappaB die during fetal development. Introduction of p50/p65-deficient fetal liver cells into lethally irradiated hosts resulted in a severe deficit of fetal liver-derived lymphocytes and their immediate precursors but an overabundance of fetal liver-derived granulocytes. Surprisingly, simultaneous transplantation of wild-type bone marrow cells rescued the production of p50/p65-deficient lymphocytes. Expression of immunoglobulin K light chains on these rescued NF-kappaB-deficient B lymphocytes was normal. These results suggest that while p50 and p65 do not regulate the maturation of pre-B cells, NF-kappaB mediates the development or survival of an early lymphocyte precursor through regulation of an extracellular factor. PMID- 9208849 TI - Regulation of the oncogenic activity of BCR-ABL by a tightly bound substrate protein RIN1. AB - RIN1 was originally identified by its ability to physically bind to and interfere with activated Ras in yeast. Paradoxically, RIN1 potentiates the oncogenic activity of the BCR-ABL tyrosine kinase in hematopoietic cells and dramatically accelerates BCR-ABL-induced leukemias in mice. RIN1 rescues BCR-ABL mutants for transformation in a manner distinguishable from the cell cycle regulators c-Myc and cyclin D1 and the Ras connector Shc. These biological effects require tyrosine phosphorylation of RIN1 and binding of RIN1 to the Abl-SH2 and SH3 domains. RIN1 is tyrosine phosphorylated and is associated with BCR-ABL in human and murine leukemic cells. RIN1 exemplifies a new class of effector molecules dependent on the concerted action of the SH3, SH2, and catalytic domains of a cytoplasmic tyrosine kinase. PMID- 9208850 TI - A kinase to remember: dual roles for MAP kinase in long-term memory. PMID- 9208851 TI - Clustering sodium channels at the node of Ranvier: close encounters of the axon glia kind. PMID- 9208852 TI - Neuregulins and their receptors: a versatile signaling module in organogenesis and oncogenesis. PMID- 9208854 TI - Neural mechanisms of visual motion perception in primates. PMID- 9208853 TI - Ca2+-triggered peptide secretion in single cells imaged with green fluorescent protein and evanescent-wave microscopy. AB - Green fluorescent protein fused to human chromogranin B or neuropeptide Y was expressed in PC12 cells and caused bright, punctate fluorescence. The fluorescent points colocalized with the endogenous secretory granule marker dopamine beta hydroxylase. Stimulation of live PC12 cells with elevated [K+], or of permeabilized PC12 cells with Ca2+, led to Ca2+-dependent loss of fluorescence from neurites. Ca2+ stimulated secretion of both fusion proteins equally well. In living cells, single fluorescent granules were imaged by evanescent-wave fluorescence microscopy. Granules were seen to migrate; to stop, as if trapped by plasmalemmal docking sites; and then to disappear abruptly, as if through exocytosis. Evidently, GFP fused to secreted peptides is a fluorescent marker for dense-core secretory granules and may be used for time-resolved microscopy of single granules. PMID- 9208855 TI - Connectin mediates adhesion in Drosophila. AB - The Drosophila cell-surface molecule connectin mediates cell-cell adhesion in vitro, and its expression pattern in vivo fits well with an adhesion role in the embryonic neuromuscular system. However, connectin mutants do not show dramatic neuromuscular defects, and ectopic expression studies so far have not supported an adhesion role. Here, we demonstrate that connectin mutants do have a phenotype; the normally connectin-positive pleural muscles fail to adhere closely together. An in vivo adhesion role is supported by misexpression studies, which result in excessive adhesion of normally connectin-negative muscles. Misexpression also causes defects in axon pathfinding. While a previous study interpreted similar defects as indicating a repulsion role for connectin, we argue that the phenotypes are consistent with connectin's adhesion role. PMID- 9208856 TI - InsP3 receptor is essential for growth and differentiation but not for vision in Drosophila. AB - Phospholipase C (PLC) is the focal point for two major signal transduction pathways: one initiated by G protein-coupled receptors and the other by tyrosine kinase receptors. Active PLC hydrolyzes phosphatidylinositol bisphosphate (PIP2) into the two second messengers inositol 1,4,5-trisphosphate (InsP3) and diacyl glycerol (DAG). DAG activates protein kinase C, and InsP3 mobilizes calcium from intracellular stores via the InsP3 receptor. Changes in [Ca2+]i regulate the function of a wide range of target proteins, including ion channels, kinases, phosphatases, proteases, and transcription factors (Berridge, 1993). In the mouse, there are three InsP3R genes, and type 1 InsP3R mutants display ataxia and epileptic seizures (Matsumoto et al., 1996). In Drosophila, only one InsP3 receptor (InsP3R) gene is known, and it is expressed ubiquitously throughout development (Hasan and Rosbash, 1992; Yoshikawa et al., 1992; Raghu and Hasan, 1995). Here, we characterize Drosophila InsP3R mutants and demonstrate that the InsP3R is essential for embryonic and larval development. Interestingly, maternal InsP3R mRNA is sufficient for progression through the embryonic stages, but larval organs show asynchronous and defective cell divisions, and imaginal discs arrest early and fail to differentiate. We also generated adult mosaic animals and demonstrate that phototransduction, a model PLC pathway thought to require InsP3R, does not require InsP3R for signaling. PMID- 9208857 TI - Surround repulsion of spinal sensory axons in higher vertebrate embryos. AB - We have tested whether the orientation of axons sprouting from bipolar dorsal root ganglion neurons is influenced by diffusible cues from surrounding tissues. Surface ectoderm, dermomyotome, and notochord exert strong chemorepulsion on axons growing in collagen gels, operating at separations beyond those found in vivo and active in cocultures of chick and mouse tissues. Basal and alar plates of the neural tube are devoid of activity, as is the posterior-half-sclerotome, which repels in a contact-dependent manner. When ganglia are sandwiched between dermomyotome and notochord placed at a distance, axon growth is channeled in a bipolar trajectory. These results show that gradients of diffusible repulsion molecules flanking axon pathways can generate linear patterns of axon growth. We suggest that such "surround repulsion" may function generally, in concert with contact-dependent guidance mechanisms, to guide axons in the developing nervous system. PMID- 9208858 TI - MAP kinase translocates into the nucleus of the presynaptic cell and is required for long-term facilitation in Aplysia. AB - Long-term facilitation of the sensory to motor synapse in Aplysia requires gene expression. While some transcription factors involved in long-term facilitation are phosphorylated by PKA, others lack PKA sites but contain MAP Kinase (MAPK) phosphorylation sites. We now show that MAPK translocates into the nucleus of the presynaptic but not the postsynaptic cell during 5-HT-induced long-term facilitation. The presynaptic nuclear translocation of MAPK is also triggered by elevations in intracellular cAMP. Injection of anti-MAPK antibodies or of MAPK Kinase inhibitors into the presynaptic cell blocks long-term facilitation, without affecting basal synaptic transmission or short-term facilitation. Thus, MAPK appears to be specifically recruited and necessary for the long-term form of facilitation. This mechanism for long-term plasticity may be quite general: cAMP also activated MAPK in mouse hippocampal neurons, suggesting that MAPK may play a role in hippocampal long-term potentiation. PMID- 9208859 TI - Mutation in the phosphorylation sites of MAP kinase blocks learning-related internalization of apCAM in Aplysia sensory neurons. AB - The synaptic growth that accompanies 5-HT-induced long-term facilitation of the sensory to motor neuron connection in Aplysia is associated with the internalization of apCAM at the surface membrane of the sensory neuron. We have now used epitope tags to examine the fate of each of the two apCAM isoforms (membrane bound and GPI-linked) and find that only the transmembrane form is internalized. This internalization can be blocked by overexpression of transmembrane constructs with a single point mutation in the two MAPK consensus sites, as well as by injection of a specific MAPK antagonist into sensory neurons. These data suggest MAPK phosphorylation at the membrane is important for the internalization of apCAMs and, thus, may represent an early regulatory step in the growth of new synaptic connections that accompanies long-term facilitation. PMID- 9208861 TI - Glutamate receptors are selectively targeted to postsynaptic sites in neurons. AB - The objective of the present study was to determine if a neuron that expresses multiple glutamate receptors targets the same receptors to all glutamatergic postsynaptic populations, or if the receptors are differentially targeted to specific postsynaptic populations. As a model for this study, we chose the fusiform cell of the dorsal cochlear nucleus. This neuron expresses multiple glutamate receptors and receives two distinct glutamatergic inputs: parallel fibers synapse on apical dendrites, and auditory nerve fibers synapse on basal dendrites. Pre- and postembedding immunocytochemistry were combined with retrograde tracing to identify the receptors expressed on postsynaptic membranes of parallel fiber and auditory nerve synapses. Most receptors were found at both populations of synapses, but the AMPA receptor subunit, GluR4, and the metabotropic receptor, mGluR1 alpha, were found only at the auditory nerve synapse. These results demonstrate that glutamate receptors are targeted to specific postsynaptic populations of glutamatergic synapses. PMID- 9208860 TI - alpha-Latrotoxin stimulates exocytosis by the interaction with a neuronal G protein-coupled receptor. AB - alpha-Latrotoxin is a potent stimulator of neurosecretion. Its action requires extracellular binding to high affinity presynaptic receptors. Neurexin I alpha was previously described as a high affinity alpha-latrotoxin receptor that binds the toxin only in the presence of calcium ions. Therefore, the interaction of alpha-latrotoxin with neurexin I alpha cannot explain how alpha-latrotoxin stimulates neurotransmitter release in the absence of calcium. We describe molecular cloning and functional expression of the calcium-independent receptor of alpha-latrotoxin (CIRL), which is a second high affinity alpha-latrotoxin receptor that may be the major mediator of alpha-latrotoxin's effects. CIRL appears to be a novel orphan G-protein-coupled receptor, a member of the secretin receptor family. In contrast with other known serpentine receptors, CIRL has two subunits of the 120 and 85 kDa that are the result of endogenous proteolytic cleavage of a precursor polypeptide. CIRL is found in brain where it is enriched in the striatum and cortex. Expression of CIRL in chromaffin cells increases the sensitivity of the cells to the effects of alpha-latrotoxin, demonstrating that this protein is functional in coupling to secretion. Syntaxin, a component of the fusion complex, copurifies with CIRL on an alpha-latrotoxin affinity column and forms stable complexes with this receptor in vitro. Interaction of CIRL with a specific presynaptic neurotoxin and with a component of the docking-fusion machinery suggests its role in regulation of neurosecretion. PMID- 9208862 TI - Beta subunits of the olfactory cyclic nucleotide-gated channel form a nitric oxide activated Ca2+ channel. AB - Cyclic nucleotide-gated channels are important in visual and olfactory transduction, and possibly in other neuronal functions. These channels have a complex permeability to Ca2+ ions that may be important in their cellular functions. They are composed of two different subunits, alpha and beta, that have been cloned and expressed, but the beta subunit alone cannot be activated by cyclic nucleotides, confounding the analysis of its characteristics. However, we found that nitric oxide can activate the homomeric expressed beta subunit, and the resulting channel possesses many properties of the L-type Ca2+ channels, including high permeability to Ca2+ ions and sensitivity to Ca2+ channel blockers. Thus, the Ca2+ permeability characteristics of native channels are mostly conferred by properties of the beta subunit, and the beta subunit alone can act as a NO-sensitive Ca2+ channel. A nearly identical conductance activated by NO is present in the membrane of rat vomeronasal neurons, indicating that homomeric beta channels exist in vivo. PMID- 9208863 TI - The efficiency of sensory information coding by mechanoreceptor neurons. AB - Most sensory systems encode external signals into action potentials for transmission to the central nervous system, but little is known about the cost or efficiency of this encoding. We measured the information capacity at three stages of encoding in the neurons of a spider slit-sense mechanoreceptor organ. For the receptor current under voltage clamp, the capacity was approximately 1400 bits/s, but when the neuron was allowed to generate a receptor potential, nonlinear membrane processes improved the capacity to >2000 bits/s. Finally, when action potentials were produced, the capacity dropped to approximately 200 bits/s, or approximately 14% of the receptor current capacity. These measurements provide a quantitative estimation of the cost of encoding analog signals into action potentials. PMID- 9208864 TI - Two distinct forms of long-term depression coexist in CA1 hippocampal pyramidal cells. AB - Two distinct forms of long-term depression (LTD), one dependent on the activation of NMDA receptors (NMDARs) and the other dependent on the activation of metabotropic glutamate receptors (mGluRs), are shown to coexist in CA1 hippocampal pyramidal cells of juvenile (11-35 day-old) rats. Both forms were pathway specific and required membrane depolarization and a rise in postsynaptic Ca2+. mGluR-LTD, but not NMDAR-LTD, required the activation of T-type Ca2+ channels, group 1 mGluRs, and protein kinase C, while NMDAR-LTD, but not mGluR LTD, required protein phosphatase activity. NMDAR-LTD was associated with a decrease in the size of quantal excitatory postsynaptic currents, whereas for mGluR-LTD there was no change in quantal size, but a large decrease in the frequency of events. NMDAR-LTD, but not mGluR-LTD, reversed NMDAR-dependent long term potentiation, and NMDAR-LTD was unaffected by prior saturation of mGluR-LTD. These findings indicate that NMDAR-LTD and mGluR-LTD are mechanistically distinct forms of synaptic plasticity. PMID- 9208865 TI - Long-term potentiation of glial synaptic currents in cerebellar culture. AB - Glial cells in the brain express neurotransmitter receptors and can respond appropriately to application of exogenous neurotransmitters such as glutamate. However, activation of receptors by endogenous, synaptically released transmitter has been difficult to demonstrate directly. Using cell-pair recording in cerebellar cultures from embryonic mouse, it is shown that activation of a cerebellar granule neuron can give rise to a rapid inward current in an adjacent glial cell. This current is mediated by activation of Ca2+-permeable AMPA/kainate receptors and is largely independent of glutamate reuptake or gap junctional coupling. Furthermore, prolonged stimulation of the granule neuron at 4 Hz can give rise to long-term potentiation (LTP) of the glial synaptic current that has similar properties to LTP of granule neuron-Purkinje neuron synaptic transmission -its induction is independent of postsynaptic depolarization, postsynaptic Ca2+ influx, or glutamate receptor activation but requires presynaptic Ca2+ influx. These findings suggest a model in which cerebellar LTP is both induced and expressed presynaptically and therefore may be detected by either neuronal or glial postsynaptic cells. PMID- 9208866 TI - Heterogeneity of release probability, facilitation, and depletion at central synapses. AB - Previous studies of short-term plasticity in central nervous systems synapses have largely focused on average synaptic properties. In this study, we use recordings from putative single synaptic release sites in hippocampal slices to show that significant heterogeneity exists in facilitation and depletion among synapses. In particular, the amount of paired-pulse facilitation is inversely related to the initial release probability of the synapse. We also examined depletion at individual synapses using high frequency stimulation, and estimated the size of the readily releasable vesicle pool, which averaged 5.0 +/- 3.0 quanta (n = 13 synapses). In addition, these experiments demonstrate that the release probability at a synapse is directly correlated with the size of its readily releasable vesicle pool. PMID- 9208867 TI - Submillisecond AMPA receptor-mediated signaling at a principal neuron-interneuron synapse. AB - Glutamatergic transmission at a principal neuron-interneuron synapse was investigated by dual whole-cell patch-clamp recording in rat hippocampal slices combined with morphological analysis. Evoked EPSPs with rapid time course (half duration = 4 ms; 34 degrees C) were generated at multiple synaptic contacts established on the interneuron dendrites close to the soma. The underlying postsynaptic conductance change showed a submillisecond rise and decay, due to the precise timing of glutamate release and the rapid deactivation of the postsynaptic AMPA receptors. Simulations based on a compartmental model of the interneuron indicated that the rapid postsynaptic conductance change determines the shape and the somatodendritic integration of EPSPs, thus enabling interneurons to detect synchronous principal neuron activity. PMID- 9208868 TI - Synergies and coincidence requirements between NO, cGMP, and Ca2+ in the induction of cerebellar long-term depression. AB - Parallel fiber synapses onto Purkinje neurons in acute cerebellar slices undergo long-term depression (LTD) when presynaptic activity coincides with postsynaptic depolarization. These electrical inputs can be respectively replaced by nitric oxide (NO) and Ca2+ photolytically released inside the Purkinje neuron, showing that these two messengers are sufficient for LTD induction. NO acts via cGMP production because inhibitors of guanylate cyclase prevent LTD but can be circumvented by photoreleased cGMP combined with Ca2+ elevation. Three inhibitors of cGMP-dependent protein kinase, Rp-8Br-PET-cGMPS, KT5823, and a novel pseudosubstrate peptide, all block LTD. LTD induction permits <10 ms gap between NO release and Ca2+ elevation, whereas 200-300 ms is allowed between uncaged cGMP and Ca2+ increase. This surprising difference in timing precision can be explained either by tighter localization and faster decay of cGMP when generated by NO rather than uncaging, or by two independent coincidence detectors in series. PMID- 9208869 TI - Recombinant alfa-interferons from naturally occurring genes. Introduction. PMID- 9208870 TI - The human interferon-alpha species and hybrid proteins. AB - Ten years of interferon (IFN) therapy have followed the approval of this agent by the US Food and Drug Administration on June 5, 1986. As the first biotherapeutic approved, IFN-alpha paved the way for the many new biotherapeutics. Regardless of this long history, however, we have just touched the surface of understanding the multitude of human IFNs. This report reviews the history of the purification of human leukocyte IFN and key aspects of our current state of knowledge of human leukocyte IFN genes and proteins. PMID- 9208871 TI - The interferon receptors. AB - During the past decade, the receptors for the type I (alpha, beta, and omega) and type II (gamma) interferons (IFNs) have been identified. The IFN-gamma receptor consists of two transmembrane chains, IFN-gammaR1 and IFN-gammaR2, both of which are required for activity. The IFN-gammaR1 chain binds the IFN-gamma ligand, whereas the IFN-gammaR2 chain is required for signal transduction. After ligand binding, Jak1 and Jak2 kinases are activated by phosphorylation and then phosphorylate the IFN-gammaR1 chain, which serves as the recruitment site for Stat1alpha (signal transducers and activators of transcription). After recruitment to the phosphorylated IFN-gammaR1 chain, Stat1alpha is then phosphorylated and released to form a Stat1alpha dimer that represents the active transcription factor for IFN-gamma-induced genes. An analogous paradigm exists for the type I IFN (IFN-alpha/beta) receptor. This receptor appears to consist of two chains, IFN-alphaR1 and IFN-alphaR2, which can be present in different forms. Thus, the IFN-alphaR1 chain is present as the full chain (IFN-alphaR1a) and as a splice-variant (IFN-alphaR1s) lacking exons IV and V; the IFN-alphaR2 chain exists in soluble, short, and long forms (IFN-alphaR2a, IFN-alphaR2b, and IFN alphaR2c, respectively). Most likely, the IFN-alphaR1a and IFN-alphaR2c chains represent the predominantly active form. After ligand binding of IFN-alpha, IFN beta, or IFN-omega species, Tyk2 and Jak1 kinases are recruited to the receptor complex and activated. The activation results in the subsequent recruitment of Stat1 (Stat1alpha and Stat1beta) and Stat2, which form a Stat1/Stat2 heterodimer after their phosphorylation. The active transcription complex IFN-stimulated gene factor-3 is formed by the association of the Stat1/Stat2 heterodimer with the p48 protein. The active IFN-stimulated gene factor-3 binds to the promoter elements of type I IFN-induced genes to initiate their transcription. Although the overall motif appears clear, there is much complexity in these interactions in that the various type I IFNs exhibit different interactions with the receptor components. Apparently, each of the IFN-alpha species exhibits a different pattern of receptor interactions that reflects their different biologic activities and will likely explain the existence of this large family of IFN-alpha species, IFN-beta, and IFN-omega that all interact with the same basal receptor. PMID- 9208872 TI - Molecular and biologic characterization of recombinant interferon-alpha2b. AB - Recombinant IFN-alpha2b (Intron A, Schering-Plough Corp, Kenilworth, NJ), derived from Escherichia coli, is currently approved worldwide for the therapy of various cancers and chronic hepatitis B and C. We describe here the purity and identity tests for both physicochemical properties and bioactivity that have been developed for the characterization of highly purified IFN-alpha2b with a specific activity of 2.5 x 10(8) IU/mg protein. The data indicate that a product of high purity can be consistently produced without DNA contamination. The antiviral bioassay chosen to measure potency is based on the ability of IFN-alpha2b to protect human foreskin fibroblast FS-71 cells from the cytopathic effects of encephalomyocarditis virus. Various immunoassays are described that have been used to quantitate the presence of binding and neutralizing antibodies in patients treated with IFN-alpha2b. Overall, the available data indicate a very low incidence of neutralizing antibody formation. We also summarize the current state of knowledge with respect to IFN reference standards for the biologic assay as well as recommendations for the calculation of antibody neutralization titers. PMID- 9208873 TI - Three-dimensional models of interferon-alpha subtypes IFN-con1, IFN-alpha8, andIFN-alpha1 derived from the crystal structure of IFN-alpha2b. AB - The crystal structure of human interferon (Hu-IFN)-alpha2b has been determined at 2.9 A resolution. This experimentally derived model provides an accurate structural scaffold on which amino acid changes between the different human IFN alpha subtypes may be compared. Accurate structural data are essential to identify structurally important residues buried in the hydrophobic core of the molecule from solvent accessible residues that may participate in receptor binding. Furthermore, the location and chemical composition of each amino acid substitution may be used to predict potential conformation changes that may occur in the different subtypes. The possible structural and surface effects of these amino acid changes on receptor binding and biologic activity are analyzed in the context of a proposed IFN-alpha receptor complex model. This model can be improved and corrected as additional biochemical and experimental structural data are obtained. These modeling techniques have been used to assess the structural and functional consequences of amino acid changes between Hu-IFN-alpha2b and consensus IFN (IFN-con1), Hu-IFN-alpha8, and Hu-IFN-alpha1, which each have distinct receptor-binding and biologic properties. Amino acids in IFN-alpha1 and IFN-alpha8 were identified that may explain the lower specific activities of these subtypes versus the activity of IFN-alpha2b. In contrast, a molecular explanation for the reported differences between IFN-alpha2b in receptor binding affinity of either IFN-alpha8 or IFN-con1 was not readily apparent. Notably, 15 of the 19 amino acid differences in IFN-con1 compared with IFN-alpha2b are located on the exterior surface, where they may enhance the antigenicity of this synthetic, nonnaturally occurring IFN. These modeling studies should assist in the design of further experiments to clarify these observations. PMID- 9208875 TI - Interacting pathways of interferon signaling. AB - Interferons (IFNs) are important modifiers of biologic response in vertebrate cells. After IFNs are induced, in response to viral or microbial infections, the IFNs bind to specific receptors on the surface of target cells and cause rapid activation of IFN-stimulated genes. These genes mediate the different biologic effects of the IFNs. The molecular characterization of IFN-induced gene activation led to the discovery of the Jak-Stat signal transduction pathway, which is now known to be shared by many other cytokines, growth factors, and hormones. Protein phosphorylation plays a key role in the Jak-Stat pathway, which consists of a cascade of specific protein-protein interactions that culminate in protein-DNA engagement and specific transcription. Although responses in the signaling pathways to various IFN-alpha species have not been examined in detail, it is likely that subtle differences in receptor interaction between the various IFN-alpha species, whether derived from natural or synthetic genes, may produce an unexpected array of biologic effects and clinical responses. PMID- 9208874 TI - Biologic activities of natural and synthetic type I interferons. AB - Because alpha-interferon (IFN-alpha) has a number of therapeutic applications in the treatment of various human cancers and diseases of viral origin, an understanding of how this family of proteins interacts with cells to induce their pleiotropic biologic activities is essential. Available data suggest that recombinant IFN-alphas from both natural and synthetic genes bind to a common cell surface receptor and induce antiviral activity in a variety of cell lines. IFN-alphas were found to differ significantly in their abilities to bind to cells; this difference varied with the types of IFN-alpha and cell type used. Consensus interferon (IFN-con1), a nonnaturally occurring synthetic IFN, and IFN alpha2b competed about equally well for receptor binding sites on Daudi and CaKi cells and were followed by IFN-alpha8 in the ability to compete. Results of affinity cross-linking experiments indicated that all three IFN-alphas interacted similarly with the multichain IFN-alpha receptor. IFN-alpha7, however, competed poorly for binding sites on both cell lines. Each of the IFN-alphas tested displayed discrete biologic differences, which also varied with the assay system used. IFN-con1 and IFN-alpha2b displayed similar antiviral activities on CaKi cells using vesicular stomatitis virus; the viral activities of these IFNs were significantly greater than those of IFN-alpha7 or IFN-alpha8. Studies with murine transfectants demonstrated significant differences in the various IFNs to interact with the IFN-alpha receptor-1 chain of the type I IFN receptor. It is yet to be established, however, that these various in vitro distinctions result in differences in clinical benefit or toxicity between the various subtypes. PMID- 9208876 TI - Effect of alpha-interferons on immune function. AB - alpha-Interferons (IFN-alphas) have been shown to have pleiotropic effects on most components of the immune system. The most studied effect of IFN-alphas on the immune system is their ability to augment natural killer cell activity, an effect that is mediated by a complex variety of mechanisms. Both naturally occurring and recombinant IFN-alphas have shown differences of as much as 100 fold in their ability to activate natural killer activity. Although consensus IFN, a synthetic, nonnaturally occurring hybrid IFN, has been reported to have a high natural killer stimulatory capacity relative to its antiviral activity, the available data and experimental design do not permit a definitive conclusion. In summary, it is clear that the complex immunomodulatory effects of IFN-alphas are likely to contribute to their therapeutic activity. PMID- 9208878 TI - The role of type I interferons in the differentiation and function of Th1 and Th2 cells. AB - T-helper cells are compartmentalized according to the cytokines they are able to produce. T-helper 1 cells produce interleukin-2 (IL-2) and interferon-gamma (IFN gamma), and develop following priming with IL-12. T-helper 2 cells produce IL-4, IL-5, and IL-10, and IL-4 is necessary to induce the differentiation of these cells. Type I IFNs have no direct effects on T-helper cell differentiation, but may regulate, especially in humans, the expression of the IL-12 receptor and thus influence T-helper 1 differentiation. On the other hand, type I IFNs can have inhibitory effects on the cytokine production by differentiated T-helper 1 cells. This suggests an important regulatory role for type I IFNs in adaptive immunity. PMID- 9208879 TI - Increasing effectiveness of interferon-alpha for malignancies. AB - Alpha-interferons (IFN-alphas) have been shown to be effective agents in inducing the regression of various malignancies, including leukemias and lymphomas as well as solid tumors, and are the first human therapeutic proteins to result in increased survival in cancer patients. Based on their pleiotropic activities, IFNs have the potential for interacting synergistically with other anticancer agents. For example, preclinical in vitro and animal data suggest a synergistic antitumor interaction between IFN-alpha2 and the antiestrogens toremifene and tamoxifen. Further studies are required to elucidate the molecular basis for such synergistic interactions, particularly with respect to IFN-stimulated genes. PMID- 9208877 TI - Induction of interleukin-1 and interleukin-1 receptor antagonist. AB - Drugs that affect the production or action, or both, of the pleiotropic inflammatory cytokine interleukin-1 (IL-1) are predicted to have a significant impact on a variety of human diseases. The biochemical properties of IL-1 and the IL-1 receptor antagonist, the mechanism for their production, and the factors that regulate their release are discussed here. The association of IL-1 with various disease states is also reviewed. Various classes of drugs that can be used to manipulate the production of IL-1 and IL- I receptor antagonist are described. Interferon-alpha2b is a potent inhibitor in vitro of the induction of IL-1 and tumor necrosis factor by IL-1 and can induce high circulating levels of the anti-inflammatory soluble tumor necrosis factor receptor and IL-1 receptor antagonist in humans. Thus, interferon-alpha2b exhibits potent anti-inflammatory effects. PMID- 9208880 TI - Does lymph node dissection still have a role in cancer therapy? PMID- 9208881 TI - Radiotherapy (RT) equipment technology in some developing countries is outdated. PMID- 9208882 TI - Introduction: paraneoplastic syndromes. PMID- 9208883 TI - Paraneoplastic syndromes: mechanisms. AB - Neoplasms may affect distant host tissues, but they always involve normal physiologic mechanisms. Van R. Potter said that "Oncogeny is blocked ontogeny," so tumors are committed to their organ anlagen. Paraneoplastic mediators are appropriate to their blocked anlagen stages. Mediators may have autocrine, paracrine, or endocrine actions. However, their release and distribution depend on the mode of cell death, and the spatial relations to host vascularity. Invasiveness and metastasis are means of normal embryonic development, and not new cancer-specific properties. Nor do human cancers acquire new foreign genetic information; thus, they cannot express neoantigens nor be recognized by the host. However, tumors breach the blood-brain barrier and basement membrane separations of ectoderm from mesenchyme and release "forbidden" self-antigens, to which host immunocytes may respond causing cell- and antibody-mediated autoimmunity. This can damage normal host tissues by a "bystander" effect. Paraneoplastic mechanisms can also be analyzed as arising from three-way interactions between cells blocked in ontogeny, their molecular messengers, and the host tissues they target. PMID- 9208884 TI - The cancer cachexia syndrome. AB - The cancer cachexia syndrome is clinically characterized by anorexia, wasting, weight loss, weakness, fatigue, poor performance status, and impaired immune function, which are unresolved by forced caloric intake. Diminished nutritional intake, maladaptive metabolic processes, and increased metabolic expenditure all play roles in the development of this syndrome. Multiple mediators of both tumor and host cell origin are mechanistic in its etiology. Treatment is not entirely satisfactory and should be directed toward improvement in the quality of life of the patient and should often include nutritional counseling. It should take into consideration both disease and treatment related factors as well as the cachexia syndrome itself. Use of progestogens (megesterol acetate, medroxyprogesterone), corticosteroids (decadron, prednisone), metoclopramide, tetrahydrocannabinol (dronabinol), and possibly anabolic steroids (nandrolone decanoate, oxandrolone), melatonin, and eicosapentaenoic acid, may yield therapeutic benefit. PMID- 9208885 TI - Fever. AB - Measurement of body temperature is the most common clinical test performed. The presence of fever is accepted as a reliable indicator of either acute of chronic disease. Although the vast majority of fevers are caused by infectious agents, some solid tumors have fever as a presenting illness usually because of associated inflammation or infection secondary to the neoplasm. However, cancer cells can spontaneously produce cytokines, small proteins with multiple biological properties. Some cytokines released by neoplastic cells are pyrogenic, ie, they produce fever directly by their action on the hypothalamic thermoregulatory center. Examples of malignant cells producing pyrogenic cytokines are renal carcinoma, lymphomas, and acute myelogenous and chronic myelogenous leukemias. The pyrogenic cytokines released by these cancers are interleukin-6, interleukin-1, tumor necrosis factor, and interferon, but others likely exist. Antipyretics reduce fever regardless of its causation; some studies suggest that fever caused by pyrogenic cytokines released from malignancies is preferentially reduced by nonsteroidal anti-inflammatory agents. PMID- 9208886 TI - Endocrine/metabolic syndromes of cancer. AB - In addition to producing symptoms directly by their mass or invasion of tissues, cancers may also make themselves evident by the secretion of cytokines, protein hormones, or hormone precursors, which in turn, results in recognizable clinical syndromes. This article reviews the endocrine syndromes of cancer, their pathophysiologic basis, and the means of diagnosis. Many of these protein hormones appear to be produced in small amounts by normal tissues where they act in paracrine fashion as cytokines. Furthermore, neoplastic transformation is associated with continued, or often dramatically amplified, production of paracrine substances, permitting them to circulate in the blood and to act as hormones. Thus, classical definitions of cytokines and hormones become blurred. In this context, so-called "ectopic" cancer production is not ectopic, but rather a modification of normal cell function. The majority of the endocrine syndromes of cancer are caused by cancer production of these cytokines or hormones. Except for cancers originating in the adrenals or gonads, cancers do not synthesize steroids and secrete them, although very rarely a cancer may metabolize a normal steroid precursor to produce a biologically active steroid. PMID- 9208887 TI - Paraneoplastic syndromes affecting the nervous system. AB - Paraneoplastic syndromes can affect virtually any portion of the nervous system. Most paraneoplastic syndromes are believed to be caused by an autoimmune reaction to an "onconeural" antigen shared by the cancer and the nervous system. The immune reaction may retard growth of the cancer, but it also damages the nervous system. Specific autoantibodies found in some individual paraneoplastic syndromes are usually associated with specific tumors. Neurological disorders, clinically and pathologically identical to paraneoplastic syndromes, may occur in some patients without cancer, but paraneoplastic antibodies are not found in these patients. The diagnosis of a paraneoplastic syndrome is based on its increased incidence in patients with cancer, the occasional response of the neurological syndrome to treatment of the underlying cancer, or the presence of specific autoantibodies. Some paraneoplastic syndromes respond to treatment of the underlying cancer or to immunosuppression but, for most syndromes, no effective treatment exists. PMID- 9208888 TI - Hematological paraneoplastic syndromes. AB - Because routine blood testing is so common in modern medicine, many cancers are first detected by the discovery of a blood test abnormality. Cancer may indirectly affect both the cellular elements of the blood as well as the coagulation system. This review of the more common hematological syndromes associated with neoplasia provides an overview of the more common entities found in clinical practice. These syndromes are important to recognize for several reasons. They provide insight into disease mechanisms and may allow earlier detection of cancer. Even if the cancer is resistant to antitumor therapy, treatment of the hematologic disorder may provide significant symptom relief. PMID- 9208889 TI - Mucocutaneous paraneoplastic syndromes. AB - Mucocutaneous paraneoplastic syndromes represent a group of dermatoses of variable morphology, pathology, and etiology that can occur as idiopathic conditions or in association with a visceral malignancy. These conditions can be categorized with respect to their predominant pathologic change: dermal "depositions," neutrophilic dermatoses, papulosquamous disorders, proliferative reactions, reactive erythemas, vacuolar degeneration of the basal layer, vasculitis, and vesiculobullous disorders. Some of these dermatoses occur more frequently in patients with hematologic malignancies whereas others are more prevalent in patients with solid tumors. The major clinical characteristics and commonly associated malignancies in patients with mucocutaneous paraneoplastic syndromes are reviewed. Suggested workups to evaluate for cancer in patients with these dermatoses are summarized. The appearance of a mucocutaneous paraneoplastic syndrome can either precede, occur concurrently with, or follow the detection of an associated neoplastic process. Therefore, the dermatosis can be the presenting feature of a previously unsuspected neoplasm or the earliest sign of recurrent cancer in an oncology patient. When the possibility of a mucocutaneous paraneoplastic syndrome is entertained, the diagnosis of the dermatosis should be confirmed either based on the clinical morphology of the lesions, the pathologic changes observed after lesional biopsy, or both. Once the diagnosis of a malignancy-associated dermatosis has been established, either an appropriate evaluation for an asymptomatic neoplasm in a cancer-free individual or an investigation for recurrence of malignancy in an oncology patient can be initiated. PMID- 9208890 TI - The association of malignancy with rheumatic and connective tissue diseases. AB - This article reviews the associations of cancer with rheumatic diseases. Recent epidemiologic data linking the autoimmune connective tissue diseases with malignancy will be emphasized. Reports linking the occurrence of malignancy with rheumatoid arthritis, systemic lupus erythematosus, idiopathic inflammatory myopathy, scleroderma, and vasculitis are described. The effect of immunomodulating drugs in the development of malignancy is discussed. Mechanisms potentially responsible for malignant transformation in the lymphoproliferation of Sjogren's syndrome are described. The relationship of gout to cancer as well as direct effects of cancer on the musculoskeletal system is also reviewed. PMID- 9208892 TI - Musculoskeletal oncology--advances in cytogenetics and molecular genetics and their clinical implications. AB - Although musculoskeletal malignancies comprise a small group of cancers, a vast number of histological subtypes have been identified attesting to the heterogeneity of this class of tumours and the growing interest in their development. The mode of management for both bone and soft tissue sarcomas has been examined extensively and treatment guidelines have been proposed. Despite the intensive study and multidisciplinary treatment, a substantial proportion of tumours remain recalcitrant to therapy and recur locally and systemically. Improved methods of characterising these tumours may help in understanding their biology. Cytogenetic and molecular genetic techniques allow a subcellular dissection of these malignancies which may aid the identification of mechanisms that are important in tumorigenesis. Already candidate genes have been isolated which may play an important role in the deregulation of proliferation and or the adoption of a malignant phenotype, features which are fundamental in tumour development. By studying the molecular biology and cytogenetics of tumours it may be possible to improve diagnostic and prognostic accuracy thereby minimising over and under treatment. PMID- 9208891 TI - Paraneoplastic syndromes of the kidney. AB - Malignant disease is associated with a wide variety of derangements in renal function and electrolyte homeostasis. In many cases this leads to a clinically significant worsening of health status and rarely may lead to the patient's death. In this review we discuss several of the important abnormalities of renal structure and function associated with neoplastic disease. PMID- 9208893 TI - Analysis of adjuvant treatment in postmenopausal patients with stage II invasive breast carcinoma--a pattern of care study and quality assurance of 431 consecutive patients in Oslo 1980-1989. AB - A retrospective study of consecutive radically operated stage II postmenopausal breast carcinoma patients diagnosed in Oslo 1980-1989 is presented. The primary aim was to analyse to which extent the increasing knowledge of the new life prolonging treatment with adjuvant tamoxifen influenced the clinical routine, and how the national recommendations were followed up. Secondary, the concomitant use of estrogen receptor (ER) analysis during the decade was to be assessed. Eligible were 150 patients without adjuvant treatment, 123 patients with radiotherapy alone, and 158 patients who had received systemic adjuvant hormone therapy. The percentage of patients without any adjuvant treatment decreased from 43% (1980 1983) to 29% (1987 1989) together with decreasing use of radiotherapy. Adjuvant tamoxifen treatment increased from 18% to 51%. In 1989, the year after publication of national recommendations of adjuvant tamoxifen treatment in stage II patients, 43% of the patients who were candidates for adjuvant tamoxifen failed to receive the therapy. Only 58% of all stage II patients had an ER analysis in the study period. The low compliance to a scientifically proven and publicly recommended adjuvant treatment of patients with stage II breast carcinoma is disappointing, as is the low performance of ER determination. To reduce the delay of implementation of new treatment modalities in the management of breast carcinoma a strict quality assurance program is needed. PMID- 9208894 TI - Implications of radiation induced apoptosis on calculated radial cell inactivation for a linear 192Ir source. AB - The cell inactivation probability as a function of the radial distance from a 4 cm linear, low dose rate (LDR) 192Ir source was calculated using a modified linear-quadratic (LQ) equation, taking into account the effect of radiation induced apoptosis. As a measure of the therapeutic range of the source, the radial distance from the midpoint of the central axis of the source to the point of 50% inactivation probability was calculated. It was found that the therapeutic range increased with increasing values of the maximal radiation induced apoptotic fraction, Fa; but only as long as the ratio between the apoptotic cell kill susceptibility, zeta, and the alpha value is larger than unity. If the ratio was smaller than unity, a decrease in therapeutic range was found. The slope of the radial cell inactivation probability curve was measured as the distance between the point of 10% and 90% inactivation probability. The slope was found to be fairly independent of the repair half-time, T(1/2), but became steeper with decreasing values of Fa. PMID- 9208895 TI - Early and long-term results of an original accelerated radiation therapy schedule in head and neck carcinoma. AB - From November 1985 until October 1988, 39 patients with head and neck carcinoma (6 patients stage I-II and 33 stage III-IV) were treated with an accelerated radiotherapy schedule designed to deliver 69.6 Gy over a period of 5 weeks. Treatment was started with 20 Gy in 10 daily fractions to sites of initial macroscopic involvement, followed by bi-fractionated radiotherapy (2 x 1.6 Gy/day) to a larger head and neck volume. Twenty patients received neo-adjuvant chemotherapy. A homolateral radical neck dissection was performed in 2 patients. Twenty-six patients (66.6%) presented with acute grade 3 complications and 5 patients (13%) with grade 4 complications. Thirteen patients (33.3%) were hospitalized for supportive care. None of the patients who were evaluated on a long-term basis developed grade 3 or 4 late complications. The 5-year loco regional control and overall survival rates were 62.4% and 33.6% respectively. Although acute toxicity is higher than in monofractionated schedules, it is manageable, and can be considered acceptable in the light of the apparently good loco-regional control thus obtained. This schedule is one of several accelerated radiotherapy programs which might merit study in prospective trials. PMID- 9208896 TI - Postoperative irradiation of laryngeal carcinoma--the prognostic value of tumour free surgical margins. AB - Between 1972 and 1987, two hundred and five carcinoma patients were treated with surgery and postoperative radiotherapy. The histological confirmation of tumour free margins was only predictive of significant differences in locoregional control in locally advanced disease. In addition, macroscopically assessed margins in advanced cases were predictive of survival probability. We suggest that the adjuvant radiotherapy was able to reduce the incidence of locoregional recurrence in the early stage microscopic positive group, hence the lack of a significant difference in the control rates. The effect on survival is therefore indirect. The advanced cases showed significantly reduced locoregional control rates and disease-specific survival times after macroscopic assessment of positive margins, possibly a sign of tumour extension beyond the margins of the radiation field. We compare our results with published reports of cases not receiving adjuvant therapy. PMID- 9208897 TI - A consecutive series of patients with laryngeal carcinoma treated by primary irradiation. AB - In Denmark there is an increasing frequency of laryngeal carcinoma, in particular in women and among these especially in supraglottic tumours. The incidence during the past 20 years has risen from about 40 to 60 cases per million per year. A series of 335 consecutive patients treated with primary radiation is presented. In one-third of all patients the tumour was localized in the supraglottic area; in women in more than half and in men in about one-fourth of the cases. The frequency of primary lymph node metastases in the supraglottic and the glottic tumours was 24% and 2% respectively. A multivariate analysis identified sex and tumour size as independent prognostic parameters of local control. Five-year survival corrected for intercurrent deaths was obtained in 59% of all cases, in 56% of supraglottic and in 92% of glottic tumours. A multivariate analysis defined localization, tumour grade and stage as independent prognostic parameters of survival. Salvage surgery was performed in about 32% of the cases, total laryngectomy in 26%, and partial laryngectomy in 6%. The survival rate among all total laryngectomies was 55%. A tracheostomy during or before radiation treatment prior to total laryngectomy had no influence on complication rate, admission time or recurrence rate. The frequency of pharyngo-cutaneous fistulae in the entire series was 11.5%; after routine use of metronidazol, however, only 5.7%. Radical neck dissection was carried out in 7.8% of the cases, by far most in the supraglottic group, only a few in the glottic carcinomas, in three-fourth in connection with a laryngectomy and in one-fourth without local recurrence in the larynx. PMID- 9208898 TI - Dosimeter gel and MR imaging for verification of calculated dose distributions in clinical radiation therapy. AB - A dosimeter gel, based on an agarose gel infused with a ferrous sulphate solution and evaluated in a magnetic resonance scanner, was used for complete verification of calculated dose distributions. Two standard treatment procedures, treatment of cancer in the urinary bladder and treatment of breast cancer after modified radical mastectomy, were examined using pixel-by-pixel and dose volume histogram comparison. The dose distributions calculated with the dose planning system was in very good agreement with the measured ones. However, in the case of the more complicated breast cancer treatment, some discrepancies were found, mainly at the beam abutment region. This may be explained by field displacements errors and by a small limitation of the dose planning utilising small electron beams in this region. The dosimeter gel system have proven to be a useful tool for dosimetry in clinical radiation therapy applications. PMID- 9208899 TI - Malignant nasopharyngeal tumours in Iceland. AB - From 1965 to 1990, 46 cases of malignant nasopharyngeal tumours were diagnosed in Iceland. The incidence rate is as low as in other Western countries, 0.6/100,000 per year. Histo-pathological diagnosis were as follows: Undifferentiated carcinoma 45%; squamous cell carcinoma 30%; non-keratinizing carcinoma 7%; and plasmacytoma 9%; lymphoma 7%; rhabdomyosarcoma 2%. Four per cent were diagnosed at stage I, 13% at stage II, 29% at stage III and 54% at stage IV. The overall crude survival at 10 years from diagnosis was 28.3%. The following factors were found to have a prognostic value: Stage of disease, size of tumour (T classification) and age at diagnosis. Nodal stage (N-classification) alone and sex were not found to be prognostic factors. There was no difference in survival among the different WHO types of cancer. Patients with carcinoma were all treated with radiotherapy. The survival of those who received more than 60 Gy was better than of those who received 60 Gy or less (p = 0.04). PMID- 9208900 TI - Human papillomavirus infection and TP53 gene mutation in primary cervical carcinoma. AB - Tumor specimens obtained from 136 patients with primary carcinoma of the uterine cervix were analyzed for the presence of human papillomavirus (HPV) sequences and for mutation of the TP53 gene. Polymerase chain reaction (PCR) showed that 130 of 136 (96%) tumors contained an oncogenic HPV 16 or 18 sequence. HPV 16 was the predominant type in cervical squamous cell carcinomas and HPV 18 was significantly associated with cervical adenocarcinomas (p < 0.05). The more dedifferentiated the primary tumor, the more frequent the HPV 16 infection and the more differentiated, the more frequent the HPV 18 infection (p < 0.05). Two out of 136 (1.5%) tumors demonstrated single-strand conformation polymorphism (SSCP) band shifts. One (positive for HPV 18) had a nonsense mutation of codon 101 in exon 4 from AAA to TAA transversion. Another (positive for L1 consensus primer set) showed a point mutation involving codon 179 in exon 5 changing CAT to CGT transition. The three specimens negative for HPV did not contain TP53 gene mutations. Our data show that mutation of TP53 is infrequent in primary cervical carcinoma and there is no inverse correlation between HPV infection and TP53 gene mutation. Other mechanisms independent of TP53 inactivation may also be implicated in tumorigenesis of the uterine cervix. PMID- 9208901 TI - Metastatic pattern at autopsy in non-resectable adenocarcinoma of the lung--a study from a cohort of 259 consecutive patients treated with chemotherapy. AB - A cohort of 259 consecutive patients with non-resectable adenocarcinoma of the lung (ACL) received chemotherapy and were followed until death with 124 cases examined by autopsy (autopsy rate 48%). Metastatic sites were identified and the following localisations were affected in 40% or more of patients post mortem: lungs, mediastinal lymph nodes, liver, pleura, adrenals, brain, and bones. Significant more metastatic sites were observed in patients who responded to the chemotherapy compared with non-responders (p = 0.04), in patients aged below the median of 58 years compared with older patients (p = 0.002), and, as expected, in patients with initial extensive disease compared with limited disease (p = 0.03). In contrast, no differences in metastatic pattern at autopsy could be detected with regard to other variables, such as initial TNM-stages, degree of histological differentiation, histologic subtypes, performance status, or LDH. PMID- 9208902 TI - Treatment of lethal midline granuloma type nasal T-cell lymphoma. AB - Nasal T-cell lymphoma of the LMG type (LMG-NTL) is characterized by progressive, unrelenting ulceration, and necrosis of the nasal cavity and midline facial tissues. The clinical behavior of this tumor in 16 patients is compared with that of a nasal lymphoma of non-LMG-NTL type (non-LMG-NTL) in 8 patients and a paranasal sinus lymphoma (PSL) in 6 patients. All patients had stage I or II disease. Fourteen of the 16 patients with LMG-NTL received chemotherapy before and/or after radiotherapy. Cause-specific 5-year survival rates for patients with LMG-NTL, non-LMG-NTL, and PSL were 22%, 75%, and 67% respectively. Seven patients with LMG-NTL, had complete response, although 3 recurred, whereas it was incomplete in 9 patients. The data indicates that it is desirable to deliver 50 Gy or more to achieve in-field control of LMG-NTL. PMID- 9208903 TI - Relationship between the type of BCR-ABL rearrangement and bone marrow histopathological features in chronic myeloid leukemia. AB - The aim of the present study was to characterize the type of BCR-ABL transcript and to correlate the molecular feature with bone marrow histology. For this purpose, we analysed the BCR-ABL rearrangement in 26 patients with chronic myeloid leukemia (CML) in the chronic phase by RT-PCR, and we also classified the bone marrow histology according to the predominance of granulocylic (GRAN) or granulocytic and megakaryocytic (GRAM/MEG) proliferation, after analysis of two independent observers. We did not find any significant difference in survival of patients presenting b2-a2 and b3-a2 transcripts or GRAN and GRAN/MEG bone marrow types, nor did we find any significant correlation of the type of BCR-ABL transcript with the bone marrow histological subgroups GRAN and GRAN-MEG (Fisher's test = 0.31). Thus, we conclude that the presence of exon b3 is not correlated to bone marrow histology in CML. PMID- 9208904 TI - Primary malignant germ cell tumours of the mediastinum--results of multimodality treatment. AB - Primary malignant mediastinal germ cell tumours are rare and considered to have poorer prognosis compared with those arising from gonads. Eighteen patients with primary mediastinal germ cell tumour were treated over an 1-year period; 9 had seminoma and 9 non-seminoma. Eight patients, 4 each with seminoma and non seminoma underwent initial tumour excision and the rest had biopsy only. All patients received cisplatin-based chemotherapy. All patients with seminoma received consolidation radiotherapy to mediastinum. Three patients with non seminoma received radiotherapy following partial response. All 9 patients with seminoma achieved complete response at the end of chemotherapy. Two patients with NSGCT had complete response to chemotherapy, 5 partial response and 2 no response. Two patients who underwent resection of the residual tumour mass are surviving free of disease. Addition of radiotherapy or second-line chemotherapy did not bring about any added response in partial and non-responders. Nine out of 9 patients with seminoma and 4/9 with non-seminoma are surviving disease-free at a median follow-up of 48 months (range 16 153 months). Mediastinal seminoma has excellent prognosis with cisplatin combination chemotherapy, whereas non-seminoma carries poor prognosis, and aggressive chemotherapy with resection of residual masses may improve the outcome. The role of additional radiotherapy and initial tumour debulking needs further evaluation. PMID- 9208905 TI - Neutral metoclopramide induces tumor cytotoxicity and sensitizes ionizing radiation of a human lung adenocarcinoma and virus induced sarcoma in mice. AB - Radiation induced cytotoxicity was potentiated by neutralized metoclopramide (nMCA; Neu-Sensamide, Oxigene Inc) when a human lung adenocarcinoma (H2981) transplanted into scid mice and an adeno-type 12 virus induced mouse sarcoma (A12B3) inoculated into CBA mice were exposed in vivo to low dose radiation at single doses of 1 and 2 Gy respectively. However, when the radiation dose was increased to 6, 10 or 18 Gy (single dose) and combined with a single dose nMCA (2 mg/kg), tumor cytotoxicity was not sensitized by the combination treatment. A fractionated dose of ionizing radiation (3 x 1 Gy) in combination with nMCA at a repeated dose of 3 x 10 mg/kg body weight (1 dose/day, i.m.) significantly increased cytotoxicity in H2981 compared with radiation given alone. nMCA alone also had a statistically significant dose dependent cytotoxic effect on H2981 growth when it was administered as repeated doses (8 doses) at 2 mg/kg or 10 mg/kg (1 dose every second day), and a similar result was achieved at 20 mg/kg but not at 2 and 10 mg/kg in the A12B3 tumor. In addition, the tumor volume at the start of treatment was important for the anti-tumor effect of nMCA (i.e. the larger initial tumor volume gave less effect on tumor growth). Taken together, our data propose that the mode of action of nMCA is different from radiation, and hence the two mechanisms are at least additive when in combination with lower radiation doses. The data further suggest that the cytotoxic mechanism is consistent with potentiating apoptosis because low and repeated doses of radiation (1-2 Gy), which are known to increase cytotoxicity by apoptosis, are sensitized by nMCA but not high doses and nMCA has more potent anti-tumor effects against H2981 tumors which have a higher constitutive apoptotic fraction of cells than A12B3. PMID- 9208906 TI - A deterministic method to calculate the radiation spectra of nuclides. AB - Recently, the computer program IMRDEC has been developed to determine the radiation spectra due to a single atomic-subshell ionisation of a stable atom by a particle, or due to the atomic deexcitation or decay of nuclides. The data needed to describe the deexcitation or decay scheme are obtained from the Evaluated Nuclear Structure Data File (ENSDF) maintained at Brookhaven National Laboratory; this results in the simplest possible input specification. The atomic data as well as the atomic relaxation probabilities are taken from the Evaluated Atomic Data Library (EADL) from Lawrence Livermore National Laboratory. The program IMRDEC calculates the radiation spectra (inclusively the atomic relaxation cascades) deterministically rather than by the Monte Carlo method; this results in much shorter calculational time per nuclide. Since many assumptions still have to be made in determining the atomic relaxation probabilities and in calculating the atomic relaxation, the deterministic method seems to be a small source of inaccuracy. PMID- 9208907 TI - Radioprotection of hematopoietic tissue by fibroblast growth factors in fractionated radiation experiments. AB - Acidic and basic fibroblast growth factors (FGF(1/2)) myeloprotect mice in single dose total body irradiation (TBI) experiments with a dose modification factor (DMF) of approximately 1.15. CFU-C assay suggests that one of the mechanisms is augmentation of the shoulder of the radiation dose response curve, and thus protection could be greater with fractionation. Four equal fractions of TBI were delivered to C3H/He mice at times 0 h, 8 h, 24 h, and 32 h. FGF(1/2) dose was 3 microg per i.v. injection given 24 and 4 hrs before the first radiation dose. FGF2 treated mice had a significant survival advantage over saline-treated mice with a DMF of 1.22 +/- 0.07 (p < 0.01). Adding a third dose of FGF2, had no additional benefit on LD(50/30) (dose of radiation lethal to 50% of animals measured at day 30) (DMF = 1.23 +/- 0.06, p < 0.01). FGF1 was not as effective with fractionation (DMF = 1.04 +/- 0.03). Increased survival in FGF2 treated mice was due to the a more rapid recovery of bone marrow hematopoietic cells and peripheral WBC, RBC and platelets. FGF2 may prove a useful treatment response modifier in clinical fractionated irradiation. PMID- 9208908 TI - Acetylsalicylic acid is unlikely to beneficially interfere with radiation-induced vasculopathy. PMID- 9208909 TI - Results of radiotherapy for T1 carcinoma of the glottis by fractions of 2.3 Gy per day. PMID- 9208910 TI - Postradiation lower motor neuron syndrome--a case report and brief literature review. PMID- 9208911 TI - Ifosfamide-based chemotherapy for recurrent or metastatic hemangiopericytoma. PMID- 9208912 TI - Intrathecal prophylaxis following autologous bone marrow transplantation for non Hodgkin's lymphoma. PMID- 9208913 TI - The emergence of Hodgkin's disease in a patient with long-standing chronic lymphocytic leukemia--case report and review of the literature. PMID- 9208914 TI - The AMP-activated protein kinase--fuel gauge of the mammalian cell? AB - A single entity, the AMP-activated protein kinase (AMPK), phosphorylates and regulates in vivo hydroxymethylglutaryl-CoA reductase and acetyl-CoA carboxylase (key regulatory enzymes of sterol synthesis and fatty acid synthesis, respectively), and probably many additional targets. The kinase is activated by high AMP and low ATP via a complex mechanism, which involves allosteric regulation, promotion of phosphorylation by an upstream protein kinase (AMPK kinase), and inhibition of dephosphorylation. This protein-kinase cascade represents a sensitive system, which is activated by cellular stresses that deplete ATP, and thus acts like a cellular fuel gauge. Our central hypothesis is that, when it detects a 'low-fuel' situation, it protects the cell by switching off ATP-consuming pathways (e.g. fatty acid synthesis and sterol synthesis) and switching on alternative pathways for ATP generation (e.g. fatty acid oxidation). Native AMP-activated protein kinase is a heterotrimer consisting of a catalytic alpha subunit, and beta and gamma subunits, which are also essential for activity. All three subunits have homologues in budding yeast, which are components of the SNF1 protein-kinase complex. SNF1 is activated by glucose starvation (which in yeast leads to ATP depletion) and genetic studies have shown that it is involved in derepression of glucose-repressed genes. This raises the intriguing possibility that AMPK may regulate gene expression in mammals. AMPK/SNF1 homologues are found in higher plants, and this protein-kinase cascade appears to be an ancient system which evolved to protect cells against the effects of nutritional or environmental stress. PMID- 9208915 TI - Molecular characterisation of integrin-procollagen C-propeptide interactions. AB - The carboxyl-terminal propeptide of type I procollagen (CPP-I) plays a key role in regulation of collagen fibrillogenesis, and may exert feedback control of collagen biosynthesis. We have previously shown that CPP-I is a ligand for the integrin alpha2beta1 [Weston, S. A., Hulmes, D. J. S., Mould, A. P., Watson, R. B. & Humphries, M. J. (1994) Identification of the integrin alpha2beta1 as a cell surface receptor for the C-propeptide of type I procollagen, J. Biol. Chem. 269, 20982-20986] suggesting that some of the phenotypic effects of C-propeptides may be mediated by adhesion receptors. Here we have extended this work to study the molecular basis of this interaction. We have broadened the ligand range by demonstrating that the C-terminal propeptide of type II procollagen supports alpha2beta1-mediated binding of NHS human fibroblasts in cell attachment assays. Also, we have used function-blocking antibodies in cell attachment and solid phase binding assays with purified integrin to expand the CPP-I receptor family, showing that integrin alpha1beta1 is also a receptor for CPP-I. Integrin alpha subunit A-domains are known to be major ligand-binding sites and recombinant alpha1 and alpha2 subunit A-domains were able to bind CPP-I. Finally we have shown that peptides corresponding to potential integrin-binding sequences in CPP I do not mediate integrin-CPP-I adhesion. Taken together, these studies indicate that the interactions between C-propeptides and integrins are more numerous than previously reported, that C-propeptides are a new class of molecule which bind to A-domains, and that the integrin-C-propeptide interaction does not utilise established peptide motifs. PMID- 9208916 TI - Evidence for channeling of intermediates in the oxidative pentose phosphate pathway by soybean and pea nodule extracts, yeast extracts, and purified yeast enzymes. AB - Evidence is presented that intermediates of the oxidative pentose phosphate pathway (OPPP) are channeled from one pathway enzyme to the next. CO2 produced from [1-14C]glucose in the presence of unlabelled pathway intermediates contained much more radioactivity than predicted by a model in which pathway-produced intermediates are in equilibrium with identical molecules in the bulk phase. This was the case whether glucose 6-phosphate (Glc6P), 6-phosphogluconolactone, or 6 phosphogluconate was added. Assumptions involved in calculating the amount of 14CO2 predicted for free mixing of 14C-labelled and unlabelled intermediates are discussed, together with the following results. (a) 14CO2 production by pea nodules in the presence of 3 mM 6-phosphogluconate was higher than in its absence. (b) Apparent channeling of intermediates was much higher for purified yeast enzymes than for yeast extract. (c) 6-Phosphogluconate and 6 phosphogluconolactone were channeled between yeast Glc6P dehydrogenase and 6 phosphogluconate dehydrogenase despite the absence of 6-phosphogluconolactonase in the purified yeast enzyme mixture. (d) When purified yeast hexokinase was physically separated from Glc6P dehydrogenase and 6-phosphogluconate dehydrogenase by a dialysis membrane, there was no apparent channeling. (e) Poly(ethylene glycol), high salt and detergents had little effect on apparent channeling of OPPP intermediates, which is consistent with a stable complex of enzymes. On the other hand, density gradient centrifugation experiments suggested a more transient interaction between the enzymes. Taken together, the results support channeling of OPPP pathway intermediates. PMID- 9208917 TI - Ribosomal protein S15 from Thermus thermophilus--cloning, sequencing, overexpression of the gene and RNA-binding properties of the protein. AB - A 6-kb DNA fragment from an extreme thermophile, Thermus thermophilus, carrying the genes for cytochrome oxidase ba3 subunit I (cbaA) and the ribosomal protein S15 (rpsO) was cloned into Escherichia coli. The gene rpsO was sequenced. The deduced amino acid sequence exhibits 59% identity to the corresponding protein from E. coli. Expression of rpsO in E. coli requires the use of a fully repressed inducible promoter because S15 from T. thermophilus is toxic for E. coli cells. When purified without denaturation from either overproducing E. coli strain or from T. thermophilus ribosomes, the S15 protein is stable and binds a cloned T. thermophilus 16S rRNA fragment (nucleotides 559-753), with low identical dissociation constants (2.5 nM), thus demonstrating that the thermophilic protein folds correctly in a mesophilic bacterium. The rRNA fragment bound corresponds in position and structure to the 16S rRNA fragment of E. coli. A similar high affinity was also found for the binding of S15 from T. thermophilus or E. coli to the corresponding E. coli 16S rRNA fragment, whereas a slightly lower affinity was observed in binding experiments between E. coli S15 and T. thermophilus 16S rRNA fragment. These results suggest that S15 from T. thermophilus recognizes similar determinants in both rRNA fragments. Competition experiments support this conclusion. PMID- 9208918 TI - 1H, 15N and 13C NMR assignments, secondary structure and overall topology of the Escherichia coli GlgS protein. AB - GlgS is a 7892-Da protein which is involved in glycogen biosynthesis in bacteria. We report the 1H, 15N and 13C NMR assignments of the backbone and side-chain resonances at 25 degrees C and pH 6.7 from two-dimensional homonuclear and three dimensional heteronuclear NMR experiments. The secondary structure of the protein was determined using sequential and medium-range NOE correlations, vicinal 3J(NH H alpha) coupling values and amide proton exchange rates. The secondary structure obtained is consistent with the secondary chemical shifts of 1H alpha, 13C alpha and 13C = O. It was found that the secondary structure of GlgS comprises two amphipathic helices (Asn10-Met21 and Glu39-Arg60), one short highly hydrophobic helix (Ile30-Val33), a short extended beta-strand-like fragment (Arg26-Asp29) and two type I beta-turns (His22-Gly25 and Thr34-Met37). An overall topology of GlgS is suggested based on long-range NOEs. The elements of secondary structure form a sandwich in which the beta-strand and the short hydrophobic helix are positioned between the two amphipathic helices. PMID- 9208919 TI - Comparative in-vivo and in-vitro 99Mo-time-differential-perturbed-angular correlation studies on the nitrogenase MoFe protein and on other Mo species of different N2-fixing bacteria. AB - Klebsiella pneumoniae, Azotobacter vinelandii and Rhodobacter capsulatus were cultivated in media containing 99MoO4(2-) . The distribution of 99Mo in cells grown under conditions of repression and derepression of nitrogenase synthesis, was investigated by anion-exchange (DEAE-Sephacel) chromatography. Cells of K. pneumoniae took up MoO4(2-) only under conditions of derepression of nitrogenase thus serving the formation of the FeMo cofactor of the MoFe protein (Kp1) as the predominant Mo-containing species. In the case of A. vinelandii, under diazotrophic growth conditions, molybdenum was preferably incorporated into the nitrogenase MoFe protein (Av1). However, if excess amounts of molybdate were present in the medium, molybdenum was also bound to the Mo-storage protein. In the presence of 20 mM NH4+, conditions which completely repress nitrogenase formation, molybdenum accumulated in the Mo-storage protein exclusively. This protein proved to be unstable towards DEAE-Sephacel, apparently releasing all the molybdenum in form of MoO4(2-) during the fractionation procedure. R. capsulatus contained, in addition to the MoFe protein (Rc1), significant amounts of other not-yet-identified Mo species, which partially are formed under conditions of both, repression and derepression of nitrogenase. The Mo centers of all these compounds were characterized by measuring the nuclear quadrupole interaction of the process 99Mo(beta-)99Tc using time differential perturbed angular correlation spectroscopy. The quadrupole coupling constant (v(Q)) determined for the Mo center in MoFe proteins was consistently in the range 66-81 MHz. The values of the coupling constants determined with intact cells and with the isolated, partially purified, MoFe proteins were in very good agreement. For the Mo-storage protein of A. vinelandii, a quadrupole coupling constant of approximately 180 MHz was determined by measurements performed with nitrogenase-repressed cells as well as with gel-filtered cell-free extracts. Our work proves that the relevant study of hyperfine interactions allows the identification of the MoFe protein and also other Mo proteins in vivo as well as in vitro. PMID- 9208920 TI - Comparison of the diphtheria mutant toxin, CRM197, with a Haemophilus influenzae type-b polysaccharide-CRM197 conjugate by optical spectroscopy. AB - A Haemophilus influenzae type-b capsular polysaccharide-CRM197 protein conjugate vaccine was compared with unconjugated CRM197 and diphtheria toxin, its parent molecule. Using CD and fluorescence spectroscopy, it has been possible to observe differences in structure and stability to pH and temperature due to the G52-->E mutation in CRM197 and the 'glycosylation' of CRM197 in the conjugate. CRM197 resembles the 'open' conformation of diphtheria toxin [Blewitt, M. G., Chung, L. A. & London, E. (1985) Biochemistry 24, 5458-5464] and the attachment of poly(ribosyl-ribitol phosphate) carbohydrate chains results in a still 'more open' state, although only a small decrease in the amount of ordered structure was observed. Fluorescence spectra of gel-filtration column fractions of the conjugate suggest that material of higher apparent molecular size is in the 'more open' conformation. Conjugated CRM197 begins unfolding at slightly lower temperatures (25-35 degrees C) than native material (> 35 degrees C). In the conjugate, tryptophan residues are more accessible to the non-ionic fluorescence quencher acrylamide at 35 degrees C. The conformational change observed at pH4-6 for diphtheria toxin is also observed for CRM197, but in the conjugate begins at higher pH. This may result from the presence of charged oligosaccharide residues on the surface or the conjugation methods used. The consequences of these changes in conformation and solution behaviour of the carrier protein in terms of its ability to induce a protective, T-cell-dependent response to H. influenzae polysaccharide remain to be determined. PMID- 9208921 TI - A partly folded state of acidic fibroblast growth factor at low pH. AB - Acid denaturation of acidic fibroblast growth factor (aFGF) at low ionic strength was monitored by far-ultraviolet circular dichroism and intrinsic fluorescence. The two spectroscopic probes displayed non-coincident transitions, which suggested the accumulation of partly folded species around pH 4.0. Although under these conditions the fluorescence of aFGF resembled that of the unfolded form of the protein, far-ultraviolet circular dichroism and proton nuclear magnetic resonance spectra indicated the presence of persistent secondary and tertiary structure. Moreover, at pH 4.0, aFGF showed cooperative thermal denaturation and interacted weakly with the hydrophobic probe N-phenyl-1-naphthylamine, showing a relatively high level of structure that did not fit into the classical molten globule category. This intermediate is also capable of interacting with liposomes and might represent a membrane translocation-competent form. PMID- 9208922 TI - Concentration of, and SDS removal from proteins isolated from multiple two dimensional electrophoresis gels. AB - We have developed a gel electrophoresis system that can concentrate proteins from spots cut out of up to 50 two-dimensional electrophoresis gels. During protein concentration, SDS is substituted with a non-ionic detergent (octyl beta glucopyranoside) which allows digestion and MS analysis of the protein directly extracted from the gel without fixation or staining. The system avoids the problems associated with the digestion of dilute protein in multiple bands by (a) greatly reducing the gel volume for digestion and thus the amount of protease required, hence lowering contamination by autodigestion products, (b) reducing the volume of solvent required for extraction of protein from the gel, thus minimising loss of material to container surfaces, and (c) removing SDS which interferes with subsequent MS or HPLC analysis. The efficiency of protein recovery ranges between an average of 80% for proteins from silver stained two dimensional gels to 90% for fluorescence and Coomassie-blue-stained gels. The method is compatible with MS analysis of very low amounts of protein from any staining system, but appears not to be useful for Edman sequencing of silver stained or fluorescent-stained proteins since the amount of N-terminal blockage appears to increase as the amount of protein isolated from the two-dimensional gel decreases. PMID- 9208923 TI - Conformation and microenvironment of the active site of xylose reductase inferred by fluorescent chemoaffinity labeling. AB - Conformation and microenvironment at the active site of xylose reductase (XR) from Neurospora crassa was probed with fluorescent chemoaffinity labeling (FCAL) using o-phthalaldehyde as a chemical initiator. Formation of a single isoindole derivative resulted in complete inactivation of the enzyme as judged by spectroscopic and fluorescence studies. Kinetic analysis of the 2,4,6 trinitrobenzenesulfonic-acid-modified XR implicated the presence of an essential lysine residue at the active site of XR. Modification of lysine in XR abolished the ability of the enzyme to form isoindole derivative, indicating that the lysine residue involved in the reaction with 2,4,6-trinitrobenzenesulfonic acid and o-phthalaldehyde is the same and that the probe o-phthalaldehyde is directed to the active site. Fluorescence studies revealed that inactivation of XR by Gdn/HCl precedes gross conformational change and the possibility of secondary conformational change was eliminated by acrylamide quenching studies. The enzyme inactivated by low concentrations of Gdn/HCl retained its ability to form the fluorescent XR-isoindole derivative indicating that inactivation is not due to conformational changes at or near the active site of XR. Gdn/HCl also had no effect on the high-affinity and low-affinity NADPH-binding sites of XR. Energy transfer experiments further revealed structural integrity at the active site of the Gdn/HCl-inactivated XR. Changes in the fluorescence emission maximum of 1 (beta-hydroxyethylthio)-2-beta hydroxyethyl isoindole (EA adduct) in solvents of varying polarity was studied, the data obtained were utilized to interpret the fluorescence behaviour of XR-isoindole derivative and assess the polarity at the active site. Experimental evidence presented here serves to suggest that the inactivation of XR by Gdn/HCl precedes conformational changes at the active site located in a microenvironment of low polarity. PMID- 9208924 TI - Expression of properly folded human glutamate decarboxylase 65 as a fusion protein in Escherichia coli. AB - Autoantibodies to the islet-cell 65-kDa variant of glutamate decarboxylase (GAD65) are found in most insulin-dependent diabetes mellitus (IDDM) patients many years before the appearance of clinical symptoms of the disease. As IDDM preventive therapies may be available in the future, an international effort is taking place to develop widely applicable anti-GAD immunochemical tests. These tests would help to detect individuals at risk before the full installation of the disease and to enroll them in prevention programs. Autoantibodies to GAD65 are mostly directed to conformational epitopes, and the enzyme is a complex molecule with a prosthetic group and 15 cysteine residues. Thus, the conformational integrity of GAD65 is essential for an appropriate anti-GAD assay. Isolation of large amounts of GAD65 from pancreas or other tissues is impractical, and no successful production of properly folded GAD65 has been reported in bacteria. Native recombinant GAD65 for immunochemical tests is usually obtained from eukaryotic expression systems. Since the large-scale production of a recombinant protein in an eukaryotic system is expensive and technically difficult, we investigated the expression of GAD65 in Escherichia coli as an alternative. A number of DNA constructs intended to export the enzyme to the periplasmic space or to improve its cytoplasmic solubility were designed and tested. Our results provide a solution to the two main problems associated with the expression of GAD65 in E. coli: misfolding, leading to the formation of inclusion bodies; and the presence of alternative initiation sites for translation that causes the preferential production of truncated variants of GAD65. We describe here the production of properly folded, fully active, and immunochemically competent GAD65 as an N-terminal fusion protein with thioredoxin. An account of the reactivity of the produced protein with sera of six IDDM patients is also presented. PMID- 9208925 TI - Structural elucidation of two polysaccharides present in the lipopolysaccharide of a clinical isolate of Burkholderia cepacia. AB - Based on the sugar composition, methylation analyses and Smith degradation, supported by NMR spectroscopic analyses and fast-atom-bombardment MS experiments, the lipopolysaccharide produced by a clinical isolate of Burkholderia cepacia was shown to contain two distinct polymers, both with linear trisaccharide repeating units; a major, containing D-rhamnose and D-galactose residues (2:1) with the structure -->3)-alpha-D-Rhap(1-->3)-alpha-D-Rhap(1-->4)-alpha-D-Galp(1 --> (major), and a minor repeating unit, constituted by D-rhamnosyl residues, with the structure -->3)-alpha-D-Rhap(1-->3)-alpha-D-Rhap(1-->2)-alpha-D-Rha p(1--> (minor). PMID- 9208926 TI - Properties of chloride-conductive pathways in rat kidney cortical and outer medulla brush-border membranes--inhibition by anti-(cystic fibrosis transmembrane regulator) mAbs. AB - The activity of the Cl(-)-conductive pathways, their regulation by protein kinase A (PKA) and their relationship to the cystic fibrosis transmembrane regulator (CFTR) protein were assessed in rat kidney cortical brush-border-membrane vesicles (cBBMV) and outer medullary vesicles (OMV) by measuring the rate of valinomycin-induced microsomal swelling by light scattering in the presence of an inward Cl- gradient. Valinomycin increased the rate of swelling of cBBMV and OMV, which is consistent with the presence of a Cl(-)-conductive pathway. PKA further increased these rates. This effect was blocked by the inhibitor of protein kinase A, suggesting that phosphorylation by PKA activates these pathways. Four anion transport inhibitors were tested ?N-phenylanthranilic acid (PhNHPhCOOH), 5-nitro 2-(3-phenylpropylamino)benzoic acid [N(PhPrNH2)BzOH], glybenclamide and 4 acetamido-4'-isothiocyanato-stilbene-2,2'-disulfonic acid?. Ph2COOH and 4 acetamido-4'-isothiocyanato-stilbene-2,2'-disulfonic acid inhibited the basal Cl( )-conductive pathways, while PKA-treated microsomes were sensitive also to N(PhPrNH2)BzOH and glybenclamide, suggesting that additional Cl- pathways were activated by phosphorylation. The pharmacological properties of these pathways were similar to those of the CFTR Cl- channel. Two anti-CFTR mAbs inhibited PKA activated valinomycin-induced swelling in cBBMV and OMV, while immunoblot analysis of the corresponding proteins with the same antibodies indicated the presence of a 170-kDa protein. The results thus indicate the presence of a PKA activated Cl(-)-conductive pathway in cBBMV and OMV, and suggest that CFTR protein is involved in PKA-activated Cl- fluxes in these vesicles. PMID- 9208927 TI - Viscoelasticity of actin-gelsolin networks in the presence of filamin. AB - Cross-linking of actin filaments by filamin by means of frequency-dependent rheology yields an increase in the filament's elasticity and stiffness. Higher cross-linker (filamin) ratios are required for mean actin-filament lengths of 5-6 microm than for random-length distribution of actin filaments. The loss modulus (i.e. the viscous portion) in the region of the internal-chain dynamics [G"(omega) approximately omega(alpha)] is influenced by the cross-linking of filaments, and with an increasing molar ratio of filamin/actin a reduction of alpha is observed. Rheological measurements reveal that actin networks are already formed at the polymerizing stage at a molar ratio of filamin/actin of less than 1:100, and electron micrographs show phase separation of actin/filament networks of low density and of actin/filament bundles. PMID- 9208929 TI - An ADP-ribosylation-factor(ARF)-like protein involved in regulated secretion. AB - A rat ADP-ribosylation factor(ARF)-like protein named ARL184 was identified by cDNA cloning. The corresponding recombinant protein had an apparent molecular mass of 22,000. The deduced amino acid sequence had 55% identity with the human ARL1 and four functional GTP-binding sites. Immunofluorescent confocal microscopy studies showed that ARL184 was present in the cytosol as well as in the Golgi apparatus, raising the possibility that it has a role in a secretory pathway. The involvement of this ARF-like protein in secretion was confirmed by demonstrating that ARL184 potentiated acetylcholine release in stably transfected PC12 cells. Collectively these results suggest that this ARL protein is a component of a regulated secretory pathway involved in Ca2(+)-dependent release of acetylcholine. PMID- 9208928 TI - Structural differences in the crossbridge head of temperature-associated myosin subfragment-1 isoforms from carp fast skeletal muscle. AB - We determined the primary structures of the three acclimation-temperature associated isoforms of myosin subfragment-1 heavy chain from fast skeletal muscle of thermally acclimated carp. These isoforms were cloned by extending 5'-regions of cDNAs that encode the rod part of myosin heavy chain specifically expressed in 10 degrees C- and 30 degrees C-acclimated carp, together with the region that encodes an intermediate structure [Imai, J., Hirayama, Y., Kikuchi, K., Kakinuma, M. & Watabe, S. (1997) J. Exp. Biol. 200, 27-34]. These three isoforms generally resembled each other in primary structure, showing 94.8, 90.9, and 92% similarity between the 10 degrees C- and intermediate-type, between the 10 degrees C- and 30 degrees C-type, and between the intermediate- and 30 degrees C-type myosin heavy chains, respectively. However, isoform-specific differences were clearly observed between the 10 degrees C- and 30 degrees C-type heavy chains in the first 60 amino acid residues from the N-terminus, where the intermediate-type showed an intermediate feature in its sequence compared to the 10 degrees C- and 30 degrees C-type isoforms. Other striking differences were observed in two surface loops between the 10 degrees C- and 30 degrees C-type isoform. Five amino acid residues out of sixteen were different in loop 1 near the ATP-binding pocket, and six out of twenty were different in loop 2 on the actin-binding site. The loops connecting beta-sheets that are known to surround the ATP-binding pocket were highly conserved in primary structure for the three types. In northern blot analysis, the accumulated mRNA levels of the 10 degrees C- and intermediate-type isoforms were significantly higher in carp acclimated to 10 degrees C and 20 degrees C than carp acclimated to 30 degrees C, whereas the level of the 30 degrees C-type isoform was significantly higher in carp acclimated to 30 degrees C than those acclimated to 10 degrees C and 20 degrees C. PMID- 9208930 TI - Identification of five new genes, closely related to the interleukin-1beta converting enzyme gene, that do not encode functional proteases. AB - Interleukin-1beta converting enzyme (ICE) was the first identified member of a growing family of cysteine proteases that now includes ten mammalian homologs. Within this large family, two functional proteins, denoted TX and TY share 60% amino-acid identity with ICE in the mature protein and, together with ICE, constitute the ICE subfamily. The present study describes the identification of five new gene sequences, denoted S1-S5, closely related to ICE and TX and belonging to this subfamily. Sequences were identified using genomic Southern blot analysis of human DNA with probes corresponding to ICE and TX exon 6. Using PCR amplification and cloning, the complete exon-6 sequence of these new genes was identified; three exhibit around 90% identity with Ice within exon 6, whereas the two others share about 70% identity with Ice. Examination of open reading frames and of amino acids essential for ICE activity indicate that none of these genes encodes for a functional protease. In conclusion, extensive analysis of the genes closely related to Ice shows that the Ice subfamily is constituted of eight members. Three of them encode for functional proteases (ICE, TX and TY) whereas the remaining members probably correspond to pseudogenes. PMID- 9208931 TI - Evolution of shorter and more hydrophilic transthyretin N-termini by stepwise conversion of exon 2 into intron 1 sequences (shifting the 3' splice site of intron 1) AB - Transthyretin cDNA was cloned from Eastern Grey Kangaroo liver and its nucleotide sequence determined. Analysis of the derived amino acid sequence of kangaroo transthyretin, together with data obtained previously for transthyretins from other vertebrate species [Duan, W., Richardson, S. J., Babon, J. J., Heyes, R. J., Southwell, B. R., Harms, P. J., Wettenhall, R. E. H., Dziegielewska, K. M., Selwood, L., Bradley, A. J., Brack, C. M. & Schreiber, G. (1995) Eur. J. Biochem. 227, 396-406], showed that the N-terminus is the region which changes most distinctly during evolution. It has been shown for human, mouse and rat transthyretins, that this region is encoded by DNA at the border of exon 1 and exon 2. Therefore, this section of transthyretin genomic DNA was amplified by PCR and directly sequenced for the Buffalo Rat, Tammar Wallaby, Eastern Grey Kangaroo, Stripe-faced Dunnart, Short-tailed Grey Opossum and White Leghorn Chicken. The splice sites at both ends of intron 1 were identified by comparison with the cDNA sequences. The obtained data suggest that the N-termini of transthyretin evolved by successive shifts of the 3' splice site of intron 1 in the 3' direction, resulting in successive shortening of the 5' end of exon 2. At the protein level, this resulted in a shorter and more hydrophilic N-terminal region of transthyretin. Successive shifts in splice sites may be an evolutionary mechanism of general importance, since they can lead to stepwise changes in the properties of proteins. This could be a molecular mechanism for positive Darwinian selection. PMID- 9208932 TI - Purification of bovine lysosomal alpha-mannosidase, characterization of its gene and determination of two mutations that cause alpha-mannosidosis. AB - Bovine kidney lysosomal alpha-mannosidase was purified to homogeneity and the gene was cloned. The gene was organized in 24 exons that spanned 16 kb and its corresponding cDNA contained an open reading frame of 2997 bp beginning from a putative ATG start codon. The deduced amino acid sequence contained a signal peptide of 50 amino acids adjacent to a protein sequence of 949 amino acids that was cleaved into five peptides in the mature enzyme; starting with the peptide derived from the N-terminal part of this precursor, their molecular masses were 35/38 (peptide a), 11/13 (peptide b), 22 (peptide c), 38 (peptide d) and 13/15 kDa (peptide e). Variation in the degree of N-glycosylation accounts for molecular mass heterogeneities of peptides a, b and e. Peptides a, b and c were disulphide-linked. A T961-->C transition, resulting in Phe321-->Leu substitution, was identified in the cDNA of alpha-mannosidosis-affected Angus cattle. In affected Galloway cattle, a G662-->A transition that causes Arg221-->His substitution was identified. Phe321 and Arg221 are conserved among the alpha mannosidase class-2 family, indicating that the substitutions resulted from disease-causing mutations in these breeds. PMID- 9208933 TI - Interactions of a bicyclic analog of colchicine with beta-tubulin isoforms alphabeta(II), alphabeta(III) and alphabeta(IV). AB - Tubulin exists as various isoforms, which differ in their assembly, drug-binding properties, and the dynamic properties of the microtubules they compose. One of the most striking differences in drug binding among the isoforms is observed with colchicine, which binds much better to the alphabeta(II) and alphabeta(IV) isoforms than to the alphabeta(III) isoform. Here we have studied the interaction of these isoforms with 2-methoxy-5-(2',3',4'-trimethoxyphenyl) tropone (MTPT), an analog of colchicine that lacks the B-ring. The kinetics of association and dissociation were studied fluorometrically, and the kinetic parameters for the two-step binding were determined for different beta-tubulin isoforms. The apparent on-rate constants for alphabeta(II), alphabeta(III) and alphabeta(IV) were 13358, 4558 and 10828 M(-1) s(-1), the off-rate constants (k(-2)) were 0.04, 0.03 and 0.02 s(-1), and the affinity constants are 3.33 x 10(5), 1.56 x 10(5) and 5.44 x 10(5) M(-1), respectively. The differences in kinetic parameters among different beta-tubulin isoforms are greatly reduced when the B-ring is removed. Our results indicate that the B-ring plays a major role in determining the isoform differences, and the results might be of importance for designing tissue specific analogs of colchicine for cancer chemotherapy. PMID- 9208934 TI - Cloning and characterization of a testis-specific, developmentally regulated A kinase-anchoring protein (TAKAP-80) present on the fibrous sheath of rat sperm. AB - cAMP is important for the initiation of mammalian sperm motility. Previously we established that a type II cAMP-dependent protein kinase is tightly associated with the fibrous sheath of rat sperm. This unique cytoskeletal structure surrounds the 9+2 axonemal network in the principal piece of the flagellum. Association of the kinase to the fibrous sheath is mediated via its regulatory subunit, RII. An RII-binding overlay procedure was used to document that RII could specifically associate with fibrous sheath polypeptides of 120 and 80 kDa. In this study, we report the cloning of a rat testis-specific, developmentally regulated, RII-binding protein (TAKAP-80). A 1.2-kb cDNA clone, isolated by screening a rat testis expression library with 32P-labeled RII, hybridized to a 1.8-kb mRNA transcript present exclusively in testis. This transcript appeared at detectable levels at 30 days after birth. Over the next 10 days the mRNA levels increased greatly. This time interval corresponds to the initiation of spermiogenesis. The complete nucleotide sequence of TAKAP-80 cDNA was obtained by polymerase chain reaction and contained a continuous open reading frame of 502 amino acids. The deduced amino acid sequence showed a clear demarcation of charged and hydrophobic amino acid residues. Amino acids 1-147 of the protein contained 45% charged residues, with lysine and arginine predominating. Similarly, amino acids 268-502 also contained a high percentage of charged amino acids (35%). In contrast, amino acids 148-267 were mostly hydrophobic and contained clusters of a repeating PXXP motif where X was predominantly valine and alanine or sometimes proline. The 1.2-kb cDNA clone was inserted into the pRSET vector and expressed as a His6 tag fusion protein in Escherichia coli. The recombinant protein was soluble and bound RIIalpha, RIIbeta and type IIalpha holoenzyme by the RII-binding overlay procedure. Deletion analysis revealed that the high-affinity interaction site for RII was contained within amino acids 258 378 of TAKAP-80. Antibodies prepared against the fusion protein recognized an 80 kDa protein present in the urea-insoluble particulate fraction of rat testis and in purified fibrous sheath preparations isolated from rat epididymal sperm. Levels of the 80-kDa immunoreactive protein were significantly higher in mature (60 days old) compared with immature (30 days old) rat testis, correlating with the mRNA levels. PMID- 9208935 TI - Sequence specificity of C-terminal Src kinase (CSK)--a comparison with Src related kinases c-Fgr and Lyn. AB - An eicosapeptide encompassing the C-terminal tail of c-Src (Tyr527) which is conserved in most Src-related protein kinases, is phosphorylated by C-terminal Src kinase (CSK) and by the two Src-related protein kinases c-Fgr and Lyn, with similar kinetic constants. Two related peptides reproducing the C-terminal segments of c-Src mutants defective in CSK phosphorylation [MacAuley, A., Okada, M., Nada, S., Nakagawa, H. & Cooper, J. A. (1993) Oncogene 8, 117-124] AFLEDSCTGTEPLYQRGENL (mutant number 28) and AFLEDNFTGTKPQYHPGENL (mutant number 29), proved a better and a much worse substrates, respectively than the wild-type peptide, with either CSK or the two Src kinases. By changing individual residues in the best peptide substrate, it was shown that the main element responsible for its improved phosphorylation is leucine at position -1 (instead of glutamine), while lysine at position -3 (instead of glutamate) has a detrimental effect, possibly accounting for the negligible phosphorylation of peptide derived from mutant number 29. By contrast to most peptide substrates, including the Src C terminal peptides, which exhibit relatively high K(m) values, a polyoma-virus middle-T-antigen-(mT)-derived peptide with tyrosine embedded in a highly hydrophobic sequence (EEEPQFEEIPIYLELLP) exhibits with CSK a quite low K(m) value (63 microM). Consistent with this, the optimal sequence selected by CSK in an oriented peptide library is XXXIYMFFF. This is different from sequences selected by Lyn (DEEIYEELX) and c-Fgr (XEEIYGIFF), although they all share a high selection for a hydrophobic residue at n-1. In sharp contrast, TPKIIB/p38syk, related to the catalytic domain of p72syk, selects acidic residues at nearly all positions, n-1 included. These data support the notion that the features determining the specific phosphorylation of the C-terminal tyrosine residue of Src do not reside in the primary structure surrounding the target tyrosine. They also show that this site does not entirely fulfil the optimal consensus sequence recognized by CSK, disclosing the possibility that as yet unrecognized CSK targets structurally unrelated to the C-terminal tyrosine residue of Src kinases may exist. PMID- 9208936 TI - Ala217 is important for the catalytic function and autoactivation of prostate specific human kallikrein 2. AB - Prostate-specific human kallikrein, hK2, is a serine protease found in prostate tissues that has 78% amino acid sequence identity with prostate-specific antigen (PSA). We have previously reported the affinity purification of hK2 heterologously expressed in a hamster cell line and demonstrated an arginine restricted substrate specificity. Here, we describe the cloning, expression, purification, and enzymatic activity of a mutant form of hK2 containing an alanine to valine substitution at residue 217 ([Val217]hK2). This mutant form was secreted into the serum-free spent media of recombinant cells as the stable proenzyme form ([Val217]phK2). Mild trypsin treatment was used to convert [Val217]phK2 to the active form, which had reduced catalytic function compared to the wild-type hK2. Kinetic studies using the chromogenic substrate D-H-Pro-Phe Arg-4-nitroanilide showed that [Val217]hK2 has significantly decreased substrate binding, with a K(m) of 4200 microM compared to 11 microM for wild-type hK2. The k(cat) for [Val217]hK2 was more than 100-fold lower than for hK2. hK2, but not [Val217]hK2, was able to activate [Val217]phK2. [Val217]hK2 also showed altered specificity on a synthetic peptide substrate compared to wild-type hK2, which exhibited partial hydrolysis at a PSA chymotrypsin-like cleavage site as well as the trypsin-like site cleaved by hK2. These results indicate that Ala217 is a key residue affecting the catalytic properties of hK2. PMID- 9208938 TI - Evolution of the enzymatic characteristics of C4 phosphoenolpyruvate carboxylase- a comparison of the orthologous PPCA phosphoenolpyruvate carboxylases of Flaveria trinervia (C4) and Flaveria pringlei (C3). AB - C4 phosphoenolpyruvate (P-pyruvate) carboxylases have evolved from ancestral C3 P pyruvate carboxylases during the evolution of C4 photosynthesis (Lepiniec et al., 1994). To meet the requirements of a new metabolic pathway, the C4 enzymes have gained distinct kinetic and regulatory properties compared to C3 enzymes. Our interest is to deduce the structure responsible for these C4-specific properties. As a model system, the orthologous ppcA P-pyruvate carboxylases of Flaveria trinervia (C4) and Flaveria pringlei (C3) were investigated by expressing them in Escherichia coli using their cDNAs. The K(m) (P-pyruvate) was about ten times higher for the C4 enzyme (650 microM) than for the C3 enzyme (60 microM). The activation by glucose 6-phosphate, which was shown by a decrease in the K(m) (P pyruvate), was about twice for the C4 enzyme and three times for the C3 enzyme. The C3 enzyme showed a very high sensitivity to L-malate with an I(0.5) (50% inhibition) value of 80 microM malate, whereas the C4 enzyme was much less sensitive with a I(0.5) value of 1.2 mM malate. To locate the structural positions responsible for these differences in kinetic and regulatory properties, chimeras of these 95% identical enzymes were made. In this study, the first 437 residues of the 966-amino-acid protein were interchanged. The results showed that the N-terminal part of the enzyme was responsible for a small but significant part of the kinetic difference observed between these two isoenzymes. Additionally, the results suggest that the N-terminus was the site for glucose 6 phosphate activation and was also responsible for the observed difference in activation by this sugar phosphate. The difference in inhibition by L-malate, however, is suggested to originate mainly from the C-terminal part of the enzyme. PMID- 9208937 TI - Role of Tyr518 and Tyr519 in the regulation of catalytic activity and substrate phosphorylation by Syk protein-tyrosine kinase. AB - The Syk protein-tyrosine kinase is expressed in many hematopoietic cells and is involved in signaling from various receptors for antigen and Fc portions of IgG and IgE. After cross-linking of these receptors, Syk is rapidly phosphorylated on tyrosine residues. We have previously reported that Syk expressed in COS cells is predominantly phosphorylated at both Tyr518 and Tyr519 at its putative autophosphorylation site. In this study, we have examined the role of each of these two residues for the catalytic activity of Syk in vitro and for the Syk induced phosphorylation of cellular proteins in intact cells. Mutation of either residue had minor effects on the catalytic activity of Syk, and even the double mutant [F518, F519]Syk was about 60% as active as the wild-type enzyme. In intact cells, however, all three mutants consistently failed to induce the extensive tyrosine phosphorylation of cellular proteins typically observed with wild-type Syk. We have recently shown that the doubly phosphorylated Y518/Y519 site is also the site for association of Syk with the SH2 domain of the Lck kinase, which suggests that although phosphates at Y518/Y519 may enhance the catalytic activity of Syk, its interaction with Src family protein-tyrosine kinases is at least equally important for the induction of downstream substrate phosphorylation. PMID- 9208939 TI - The role of the 90-kDa heat-shock protein and its associated cohorts in stabilizing the heme-regulated eIF-2alpha kinase in reticulocyte lysates during heat stress. AB - The heme-regulated eIF-2alpha kinase (HRI) is activated not only in heme deficient rabbit reticulocyte lysates (RRL), but also in hemin-supplemented RRL treated with heat-shock, N-ethylmaleimide (MalNEt) or heavy metal ions. We have demonstrated previously that heat-shock proteins, Hsp90, Hsp70 and FKBP52, are associated with HRI in RRL; the association of HRI with Hsp90 and FKBP52, but not Hsp70, is enhanced by hemin. To study the role of Hsp90 and its associated cohorts in the regulation of HRI, we examined the interaction of these proteins with HRI in hemin-supplemented RRLs during heat or oxidative stress. The association of HRI with Hsp90, FKBP52 and p23 was maintained in heat-, MalNEt- or Hg2(+)-treated hemin-supplemented RRL. Glycerol gradient centrifugation and gel filtration on Sephacryl S-300 indicated that neither heat shock nor MalNEt treatment affected the apparent molecular mass of HRI in hemin supplemented RRL. In addition, active HRI was coimmunoprecipitated with 8D3 anti-Hsp90 from both heme-deficient and MalNEt-treated hemin-supplemented RRL. These results demonstrate that activation of HRI in response to heat stress and oxidative stress does not require dissociation of Hsp90 from HRI. Furthermore, HRI activity was inhibited upon addition of hemin to Hsp90-depleted heme-deficient RRL, indicating that inhibition of HRI activity by hemin is not mediated by the reassociation of Hsp90 with HRI. We also examined the dynamics of the interaction of Hsp90 with HRI. Reconstitution of the interaction of Hsp90 with HRI was stimulated by elevated temperature and required both Mg2+ and ATP. Addition of purified Hsp90 to hemin-supplemented RRL which had been treated with MalNEt to inactivate its capacity to chaperone protein renaturation, protected HRI from irreversible denaturation and aggregation upon incubation at 41 degrees C. Our results suggest that Hsp90 interacts with HRI primarily in its capacity as a molecular chaperone, stabilizing HRI from denaturation under conditions of heat stress and oxidative stress. PMID- 9208940 TI - Mechanism of inactivation of a catalytic antibody by p-nitrophenyl esters. AB - Antibody CNJ206 catalyses the hydrolysis of p-nitrophenyl esters with significant rate enhancement; however, after a few cycles, 90% of the catalytic activity of CNJ206 is irreversibly lost. This report investigates the properties of the inactivated Fab (fragment antigen binding). After inactivation, the residual esterase activity of CNJ206 is similar to that of the catalytic antibody inhibited by the transition-state analogue (TSA) used to elicit it; the affinity of CNJ206 for the TSA is also dramatically lowered. Here we propose a simple scheme that accounts for the steady-state kinetics of inactivation. The following lines of evidence, when taken together, suggest that stable acylated tyrosine side chains within or close to the Fab combining site are involved in the inactivation process: isoelectric focusing and matrix-assisted-laser-desorption ionisation-time-of-flight (MALDI-TOF) mass spectrometry show that incubation with substrate results in several acylated Fab species; inactivation is stable at pH 8, is reversed by mild hydroxylamine treatment and follows the same kinetics as inhibition of binding, which is slowed down by the presence of the TSA hapten. Analysis of the Fab-TSA X-ray structure shows that three tyrosine residues are potential candidates for the inactivation of CNJ206 by its substrates, Tyr L96 being the most likely one; this also suggests that site-directed mutation of one or more of these residues might prevent substrate inactivation and significantly improve catalysis. PMID- 9208941 TI - Demonstration of the presence of alpha-1,6-branched side chains in the mannan of Candida stellatoidea. AB - A mild acetolysis of the mannans of Candida stellatoidea was performed after acetylation to yielded an alpha-1,6-branched mannohexaose, the presence of which had been predicted from the appearance of a specific H1-H2-correlated cross-peak in two-dimensional homonuclear Hartmann-Hahn spectroscopy. In this study, we found that the de-O-acetylation of a 4-O-acetyl group at the branching point, the 3,6-di-O-substituted mannose unit, of an acetylated oligosaccharide by sodium methoxide is significantly slower than that of other acetyl groups. We could separate the 4-O-acetylated branching oligosaccharide from linear isomer using high-performance liquid chromatography. Before and after the de-O-acetylation of the purified branching oligosaccharide, their 1H-NMR signals were sequentially assigned by means of the nuclear Overhauser effect. In the sequential NMR assignment study, we showed that the alpha-1,6-linked mannose unit is attached to the 3-O-substituted unit based on the presence of NOE cross-peak between H1 of the branching mannose unit and H6 of the 3-O-substituted mannose unit. An enzyme linked immunosorbent inhibition assay of the reactivity of factor 4 serum to C. stellatoidea mannan by several oligosaccharides indicated that the alpha-1,6 branched oligosaccharide and the beta-1,2 linkage-containing oligosaccharides showed inhibitory activity. This result indicates that factor 4 serum, as well as factor 5 and 6 sera, contains antibodies against beta-1,2-linked mannose units which have been reported to participate in pathogenicity via cytokine production and/or adherence. From the assignment results of H1-H2-correlated cross-peaks of oligosaccharides and mannans, the molar ratio of the mannan side chains was proposed. In this study, we demonstrated that the epitope structure of the C. stellatoidea type I strains was the same as that of the C. albicans NIH B-792 (serotype B) strain. PMID- 9208942 TI - A point mutation responsible for defective function of the aryl-hydrocarbon receptor nuclear translocator in mutant Hepa-1c1c7 cells. AB - A 3,4-benzopyrene-resistant mutant clone (c4) of the mouse hepatoma Hepa-1c1c7 cell line was examined for the mutation that causes the defective function of aryl-hydrocarbon receptor (AHR) nuclear translocator (Arnt). Arnt dimerizes with AHR and mediates the induction signal of aryl-hydrocarbon hydroxylase activity. The Arnt cDNAs of c4 cells were cloned by reverse-transcription/PCR to compare the sequences with that of wild-type Arnt cDNA. The Arnt cDNA of c4 cells was found to have a single point mutation, leading to replacement of Gly326 with Asp between two internal repeats in the highly conserved Per-Arnt-Sim (PAS) domain, PAS A and PAS B. The inability of [Asp326]Arnt/AHR heterodimers to enhance reporter gene transcription under the control of the CYP1A1 gene promoter and enhancer confirmed that the G326-->D substitution was a causative mutation. While fluorescence microscopy and coimmunoprecipitation experiments showed that this mutant form of Arnt was not changed from wild-type Arnt in terms of nuclear localization or heterodimer formation with AHR, the binding activity of the [Asp326]Arnt x AHR heterodimer to the xenobiotic-responsive element was reduced markedly. Determination of the turnover rate in COS-7 cells transfected with expression plasmids for mutant Arnt or normal Arnt showed that the mutant protein turned over with an accelerated rate compared with that of the normal. Moreover, the mutant protein displayed increased proteolytic digestibility in vitro with various proteases. PMID- 9208943 TI - Anti-insect toxin 5 (AaIT5) from Androctonus australis. AB - An insect-selective scorpion toxin (AaIT5) was purified from the venom of the North African scorpion Androctonus australis, and its amino acid sequence was determined by a combination of automated Edman degradation, electrospray ionization mass spectrometry, and sequence alignment. This insect toxin is very potent against the tobacco budworm, Heliothis virescens (100% lethal dose < 1.8 microg/100 mg body mass) and shows a distinct insect specificity and various symptoms. It is not toxic to mice after subcutaneous injection. The molecular mass of this toxin is 6882 Da and the amino acid sequence is similar to those of Androctonus australis anti-insect toxin 4 (AaIT4), Leiurus quinquestriatus depressant anti-insect toxins (LqhIT2, LqqIT2), and Buthotus judaicus depressant anti-insect toxin (BjIT2). PMID- 9208944 TI - Widespread tissue expression of gastrin-binding-protein mRNA. AB - Glycine-extended forms of gastrin (gastrin-Gly) are thought to be involved in the autocrine growth control of colorectal carcinomas. The recently described gastrin binding protein has been suggested to be a gastrin-Gly accepting receptor. Northern blot analysis demonstrated the expression of gastrin-binding-protein mRNA in many tissues of mouse, rat, and man. The gastrin-binding-protein mRNA expression was confirmed by reverse-transcribed PCR analysis. Analysis of the cDNA and the deduced amino acid sequence of the PCR-amplified rat gastrin-binding protein DNA fragments revealed sequence identity (except in a single position) with the corresponding human and pig gastrin-binding protein and with the alpha subunit of a rat and human mitochondrial trifunctional enzyme, involved in fatty acid oxidation. The widespread and abundant tissue expression of gastrin-binding protein mRNA and its sequence identity with a fatty-acid-oxidizing enzyme do not support the view that it represents a genuine gastrin receptor. PMID- 9208945 TI - DNA interactions of bifunctional dinuclear platinum(II) antitumor agents. AB - Modifications of natural DNA in a cell-free medium by dinuclear bisplatinum complexes with equivalent coordination spheres, represented by the general formula [?trans-PtCl(NH3)2?2(H2N-R-NH2)]2+, where R is a propane or hexane, were studied by various methods of biochemical analysis or molecular biophysics. These methods include binding studies by means of differential-pulse polarography, measurements of melting curves with the aid of absorption spectrophotometry, measurements of CD spectra, ELISA with specific antibodies that recognize DNA modified by platinum complexes, interstrand cross-linking assay employing gel electrophoresis under denaturing conditions and mapping of DNA adducts by means of transcription assays. The results indicated that the major adduct of [?trans PtCl(NH3)2?2(H2N-R-NH2)]2+ in DNA was an interstrand cross-link which was formed with a relatively short half-time (approximately 1 h). At least some types of these interstrand cross-links induced local denaturational changes in the DNA. The results of analyses of interactions of [?trans-PtCl(NH3)2?2(H2N-R-NH2)]2+ with linear DNA at relatively higher levels of the modification could be interpreted to mean that these dinuclear platinum complexes were also capable of intrastrand-cross-link formation between adjacent base residues in DNA. However, these intrastrand adducts of [?trans-PtCl(NH3)2?2(H2N-R-NH2)]2+ distorted DNA conformation in a way different from the DNA intrastrand adducts of cisplatin. In addition, the DNA adducts of the dinuclear platinum complexes inhibited DNA transcription in vitro. The length of the aliphatic linker chain affected the DNA binding mode of [?trans-PtCl(NH3)2?2(H2N-R-NH2)]2+ and the resulting conformational changes in DNA. The extensive analysis of DNA interactions with [?trans-PtCl(NH3)2?2(H2N-R-NH2)]2+ described in this communication has provided further experimental support for previous suggestions [Farrell, N. (1991) in Platinum and other metal coordination compounds in cancer chemotherapy (Howell, S. B., ed.) pp. 81-91, Plenum Press, New York] that the binding of the dinuclear platinum complexes modifies DNA in a way that is different from the modification by antitumor cisplatin. Thus, the results of this work are consistent with the hypothesis that platinum drugs that bind to DNA in a manner fundamentally different from that of cisplatin can exhibit altered biological properties, including a different spectrum and intensity of antitumor activity. PMID- 9208946 TI - Identification of cDNAs encoding sterol methyl-transferases involved in the second methylation step of plant sterol biosynthesis. AB - Two methyl transfers are involved in the course of plant sterol biosynthesis and responsible for the formation of 24-alkyl sterols (mainly 24-ethyl sterols) which play major roles in plant growth and development. The first methyl transfer applies to cycloartenol, the second one to 24-methylene lophenol. Five cDNA clones encoding two Arabidopsis thaliana, two Nicotiana tabacum and one Ricinus communis S-adenosyl-L-methionine (AdoMet) sterol methyltransferases (SMT) were isolated. The deduced amino acid sequences of A. thaliana and N. tabacum SMT are about 80% identical in all possible combinations. In contrast they are about 40% identical with the deduced amino acid sequence of R. communis SMT and the published Glycine max sequence. Both A. thaliana and one N. tabacum SMT cDNAs were expressed in a yeast null mutant erg6, deficient in AdoMet zymosterol C24 methyltransferase and containing C24-non-alkylated sterols. In all cases, several 24-ethylidene sterols were synthesized. A thorough study of the sterolic composition of erg6 expressing the A. thaliana cDNA 411 (erg6-4118-pYeDP60) showed 24-methylene and 24-ethylidene derivatives of 4-desmethyl, 4alpha-methyl and 4,4-dimethyl sterols as well as 24-methyl and 24-ethyl derivatives of 4 desmethyl sterols. The structure of 5alpha-stigmasta-8, Z-24(24(1))-dien-3beta ol, the major sterol of transformed yeasts, was demonstrated by 400 MHz 1H NMR. Microsomes from erg6-4118-pYeDP60 were shown to possess AdoMet-dependent sterol-C methyltransferase activity. Delipidated preparations of these microsomes converted cycloartenol into 24-methylene cycloartanol and 24-methylene lophenol into 24-ethylidene lophenol, thus allowing the first identification of a plant sterol-C-methyltransferase cDNA. The catalytic efficiency of the expressed SMT was 17-times higher with 24-methylene lophenol than with cycloartenol. This result provides evidence that the A. thaliana cDNA 411 (and most probably the 3 plant SMT cDNAs presenting 80% identity with it) encodes a 24-methylene lophenol C-24(1) methyltransferase catalyzing the second methylation step of plant sterol biosynthesis. PMID- 9208947 TI - Sequence of a gene cluster from Klebsiella pneumoniae encoding malonate decarboxylase and expression of the enzyme in Escherichia coli. AB - Malonate decarboxylase of Klebsiella pneumoniae consists of four different subunits and catalyzes the conversion of malonate plus H+ to acetate and CO2. The catalysis proceeds via acetyl and malonyl thioester residues with the phosphribosyl-dephospho-CoA prosthetic group of the acyl carrier protein (ACP) subunit. From a cosmid library of K. pneumoniae, a gene cluster of 9 kb has been isolated and sequenced that included the structural genes for the malonate decarboxylase. The cluster consisted of the eight consecutive genes mdcABCDEFGH and the divergently oriented mdcR gene. The intergenic regions were short (usually < 17 bp, 136 bp between mdcE and mdcF) and ribosome binding sites were found 4-10 bp before each gene. According to N-terminal protein sequencing, the mdcA, C, D and E genes encoded subunits alpha, delta, beta and gamma of malonate decarboxylase. Data bank searches for related proteins with known function revealed that MdcA represents the ACP-transferase and that MdcD and E together probably function as malonyl-S-ACP decarboxylase. MdcC is the (apo) ACP subunit. MdcB and MdcG could be involved in the synthesis and attachment of the prosthetic group. MdcH is similar to various malonyl-CoA:ACP-SH transacylases and therefore probably involved in the initial activation of the enzyme by malonylation. MdcF is a membrane protein that could function as a malonate carrier. The mdcR gene encodes a protein of the LysR regulator family. Malonate decarboxylase was functionally expressed in Escherichia coli from plasmids harbouring the entire gene cluster including mdcR. As partial deletion of the mdcR gene impaired growth of the transformants on malonate, MdcR is probably a transcriptional regulator of the mdc genes. PMID- 9208948 TI - General structure/function properties of microbial methionyl-tRNA synthetases. AB - Alignment of the sequences of methionyl-tRNA synthetases from various microbial sources shows low levels of identities. However, sequence identities are clustered in a limited number of sites, most of which contain peptide patterns known to support the activity of the Escherichia coli enzyme. In the present study, site-directed mutagenesis was used to probe the role of these conserved residues in the case of the Bacillus stearothermophilus methionyl-tRNA synthetase. The B. stearothermophilus enzyme was chosen in this study because it can be produced as an active truncated monomeric form, similar to the monomeric derivative of E. coli methionyl-tRNA synthetase produced by mild proteolysis. The two core enzyme molecules share only 27% identical residues. The results allowed the identification of the binding sites for ATP, methionine and tRNA, as well as that responsible for the tight binding of the zinc ion to the enzyme. It is concluded that the thermostable synthetase adopts a three-dimensional folding very similar to that of the E. coli one. Therefore, the two methionyl-tRNA synthetase sequences, although significantly different, maintain a common scaffold with the functionally important residues exposed at constant positions. Sequence alignments suggest that the above conclusion can be generalized to the known methionyl-tRNA synthetases from various sources. PMID- 9208949 TI - Biochemical characterization of purified, human recombinant Lys304-->Glu medium chain acyl-CoA dehydrogenase containing the common disease-causing mutation and comparison with the normal enzyme. AB - Recombinant, normal human medium-chain acyl-CoA dehydrogenase (MCADH) and the common, human disease-causing K304E mutant ([Glu304]MCADH) protein were expressed in Escherichia coli using an optimized system, and the enzymes were purified to apparent homogeneity. The crucial factor leading to the production of active [Glu304]MCADH protein is the expression in E. coli cells at reduced temperature (28 degrees C). Expression in the same system at 37 degrees C results in very low amounts of active mutant protein. Several catalytic and physicochemical parameters of these two proteins have been determined and were compared to those of purified pig kidney MCADH. Although [Glu304]MCADH has approximately the same rate of substrate reduction with dodecanoyl-CoA and the same V(max) as human MCADH with the best substrate for the latter, octanoyl-CoA, the K(m) in the mutant MCADH is fourfold higher, which generates a correspondingly lower catalytic efficiency. Importantly, V(max) obtained using the natural acceptor, electron transfer flavoprotein, is only a third that for human MCADH. The V(max)/K(m) versus chain-length profile of the mutant shows a maximum with dodecanoyl-CoA which differs markedly from that of human MCADH, which has maximal efficiency with octanoyl-CoA. The substrate specificity of the mutant is broader with a less pronounced activity peak resembling long-chain acyl-CoA dehydrogenase. The purified mutant enzyme exhibits a reduced thermal stability compared to human wild-type MCADH. The major difference between the two proteins expressed in E. coli is the more pronounced lability of the K304E mutant in crude extracts, which suggests a higher susceptibility to attack by endogenous proteases. Differences between tetrameric [Glu304]MCADH which survives the first step(s) of purification and corresponding MCADH are minor. The overall differences in properties of [Glu304]MCADH together with its impaired folding and tetramer assembly may contribute to the generation of the abnormalities observed in patients homozygous for the K304E mutation. PMID- 9208950 TI - Specific interaction of the Streptomyces chitin-binding protein CHB1 with alpha chitin--the role of individual tryptophan residues. AB - Streptomyces olivaceoviridis secretes a so far unique protein of 18.7 kDa (CHB1) which lacks catalytic activity. It interacts highly specifically with alpha chitin, but not with beta-chitin, chitosan, or cellulose. Each of the five codons for tryptophan (Trp) in the chb1 gene was replaced by those for leucine (Leu) or tyrosine (Tyr). Eight corresponding mutant proteins and the wild-type protein were purified to homogeneity and their binding capacity to alpha-chitin was determined. The relative affinities to anti-CHB1 antibodies, the kinetics of binding, the dissociation constants, circular dichroism, and fluorescence emission spectra for three mutant types were compared to the characteristics of CHB1. The presented data lead to the following conclusions. (a) CHBI presents a highly flexible protein lacking alpha-helices. (b) Replacement of each of the buried Trp residues (Trp134 and Trp184) leads to conformational alterations and, in due course, to a considerably reduced binding affinity of the protein. (c) The exchange of the exposed Trp 57 by either Leu or Tyr results in relatively slight topological changes, but entails a loss of binding capacity of about 90%. (d) The dissociation constant was highest for the mutant protein [L57]CHB1 (2.17 microM), followed by [L134]CHB1 (0.91 microM) and [L184]CHB1 (0.26 microM), and lowest for the progenitor CHB1 (0.11 microM), indicating its strong affinity to the unsoluble substrate. (e) The data suggest that the exposed Trp57 contributes directly and significantly to the interaction of CHB1 with alpha-chitin. PMID- 9208951 TI - Structural studies of the O-antigenic polysaccharide from Escherichia coli O167. AB - The structure of the O-antigenic polysaccharide from Escherichia coli O167:H5 has been investigated. Sugar and methylation analyses, fast-atom-bombardment mass spectrometry and 1H- and 13C-NMR spectroscopy were the main methods used. The structure of the repeating unit of the polysaccharide was found to be: [formula in text]. Oligosaccharide derivatives of the polysaccharide were obtained by HF solvolysis and by a Smith degradation. Furthermore, base treatment of the polysaccharide led to a degraded polymeric material. For the methylated polysaccharide the amide linkage between alanine and the galacturonic acid residue was reductively cleaved with LiBD4 in ethanol, to give, among other things, a 3-O-methyl galactose derivative. PMID- 9208952 TI - Thermodynamic significance of the lactate gradient. AB - The theory that some bacteria can save energy by an energy-recycling process, in which protons are excreted with metabolic end-products with variable stoichiometry, has been examined by 1H-NMR. A method has been developed that utilises observed differences in the Hahn T2 relaxation of metabolites in the intracellular and extracellular compartments to distinguish and quantify metabolite signals originating from both compartments. It was found that the lactate electrochemical-potential gradient calculated from the fraction of lactate that is sufficiently mobile to contribute to the NMR signal was in exact balance with the proton electrochemical-potential gradient over a wide range of pH values. The conclusion was reached that previous reports of variable stoichiometry were due to 'bound' lactate at high intracellular pH that could neither contribute neither to the NMR signal nor to the lactate electrochemical potential gradient. PMID- 9208953 TI - Stomal ulcer after gastric bypass. AB - BACKGROUND: Stomal ulcer is a serious complication of gastrogastric fistula following Roux-en-Y gastric bypass for obesity. STUDY DESIGN: A 1-8 year continuous followup of 499 patients with gastric bypass in continuity (GB) and isolated gastric bypass (IGB) documented the incidence of fistula formation, development of stomal ulcer, stimulation of acid production within the gastric pouch, and response to treatment. RESULTS: In 123 GB patients, staple line disruption occurred in 36 (29%) and stomal ulcer occurred in 20 (16%). Gastrogastric fistula with stomal ulcer was significantly lower in 376 patients who underwent IGB, (ie, 11 patients [3%]). Significantly larger amounts of acid, a lower pH, and a greater time with a pH less than 2 were found in the gastric pouches of patients who developed stomal ulcer after Roux-en-Y gastric bypass. All patients had a perforated staple line. Successful closure of the staple line significantly decreased acid production and pH in the gastric pouch when tested before and after remedial operation with healing of stomal ulcers. CONCLUSIONS: Stomal ulcer after gastric bypass is the result of acid production in the bypassed stomach in the presence of a gastrogastric fistula. Separation of the gastric pouch from the main stomach decreases the incidence of fistula formation and stomal ulcer but does not eliminate it. Interposition of a well vascularized organ, the jejunum between the pouch and main stomach, is an attractive solution for patients who require remedial operations on the stomach and possibly for primary operations as well. PMID- 9208954 TI - The accumulated experience of the Israeli Advanced Trauma Life Support program. AB - BACKGROUND: Between January 1990 and May 1995 one faculty in Israel taught Advanced Trauma Life Support (ATLS) courses to 3,700 physicians. Two types of courses were given to three subpopulations. We studied the influence of demographic variables on students' achievements in the course and compared students' achievements as a function of their course type. STUDY DESIGN: This study was conducted as a concurrent longitudinal study. RESULTS: Achievements of 3,700 students were analyzed. The precourse grade, type of course, and their interaction were found to have a significant effect on the postcourse grades. Physicians practicing surgical subspecialties, in general, did better, as did students educated in English-speaking countries. Students who took part in the Combat Trauma Life Support (CTLS) course, which included the entire ATLS course and additional lectures and exercises, also ended with better scores. CONCLUSIONS: Physician's country of origin and clinical subspecialty have a significant effect on the cognitive achievement in the ATLS course provided in Israel. An expanded ATLS course (CTLS), to include additional military trauma topics as well as additional skill station training, can improve the results of the postcourse grades. PMID- 9208955 TI - Granulomatous appendicitis: Crohn's disease, atypical Crohn's or not Crohn's at all? AB - BACKGROUND: Crohn's disease isolated to the appendix has primarily been documented in case reports. We contribute a series with longterm followup and a literature review. STUDY DESIGN: A retrospective review of 1,133 consecutive appendectomy specimens over the 6-year period ending in 1994 identified seven patients with isolated granulomatous appendicitis. Two patients presented before the review period. These nine patients are reviewed and 156 patients identified in the world literature. RESULTS: Granulomatous appendicitis usually presents as an indolent course of appendicitis. No patient developed enterocutaneous fistula after appendectomy in our series. A mean followup of 7.3 years in our patients revealed no evidence of Crohn's disease. CONCLUSIONS: Granulomatous inflammatory disease isolated to the appendix differs from typical Crohn's disease with a decreased occurrence of enterocutaneous fistulas and rare recurrence. Consequently, isolated granulomatous appendicitis without small bowel or cecal involvement may not represent true Crohn's disease. Patients can be treated with minimal morbidity by appendectomy alone. If isolated granulomatous appendicitis does represent Crohn's disease, its longterm course in the majority of patients is extremely benign. PMID- 9208956 TI - Incidence, risk factors, and treatment of pancreatic leakage after pancreaticoduodenectomy: drainage versus resection of the pancreatic remnant. AB - BACKGROUND: Pancreatic leakage is a major cause of morbidity and mortality after pancreaticoduodenectomy, with incidences varying between 6-24% and a mortality rate up to 40%. Treatment is an issue of controversy. In this study we analyzed risk factors for pancreatic leakage and the results of early resection of the pancreatic remnant versus drainage procedures for leakage of the pancreaticojejunostomy. STUDY DESIGN: From 1983 to 1995, 269 patients underwent pancreaticoduodenectomy, with pancreaticojejunostomy. Patients with manifestations of pancreatic leakage were compared with nonleakage patients to evaluate risk factors. Patients with leakage were divided into two treatment groups. One group comprised patients undergoing percutaneous or surgical drainage procedures; the other had patients undergoing resection of the pancreatic remnant. RESULTS: Twenty-nine patients (11%) had clinical manifestations of pancreatic leakage, and the mortality in these patients was 28% (overall mortality: 3.7%). Leakage occurred after a median of 5 days (range 1-20). Age, preoperative bilirubin level, and albumin counts were not risk factors for pancreatic leakage. Small pancreatic duct size (< 2 mm) (p < 0.01) and ampullary carcinoma as histopathologic diagnosis (p < 0.05) were risk factors. The median number of relaparotomies was two (range 0-4) in the drainage group (n = 21), versus 1.5 (range 1-5) in patients who underwent resection (n = 8). The median hospital stay was 74 days (range 36-219), versus 55 days (range 22-107) for the drainage and resection groups, respectively (p < 0.05). Mortality was lower in patients who underwent resection, 38 versus 0% (p < 0.05). CONCLUSIONS: Leakage of the pancreatic anastomosis is a severe complication after pancreaticoduodenectomy and carries a high mortality rate (28%). Completion pancreatectomy could be performed without additional mortality. In patients with severe and persistent leakage of the anastomosis, early completion pancreatectomy is the treatment of choice. PMID- 9208957 TI - No-cut thoracoscopic lung plication: a new technique for lung volume reduction surgery. AB - BACKGROUND: Lung volume reduction surgery (LVRS) using a linear cutting stapler or laser ablation via median sternotomy or thoracoscopy is a current therapy for symptomatic emphysema. The primary causes of morbidity and mortality (as high as 20%) are existing comorbidities and prolonged air leaks secondary to visceral pleural division. We report a novel technique using minimally invasive techniques designed to achieve volume reduction while preserving the visceral pleura. A novel lung grasper and a knifeless stapler are used to permanently plicate lung tissue without cutting visceral pleura. STUDY DESIGN: This prospective analysis involves a consecutive series of patients who had LVRS using this method. Between May 1995 and September 1996, 32 patients underwent 50 unilateral, staged bilateral, or bilateral thoracoscopic lung plication procedures. The indications for LVRS were standard; they included severe limiting dyspnea (forced expiratory volume in one second [FEV1] = 0.68 +/- 0.05), hyperinflated lungs with flattened diaphragms on chest x-ray, and diffuse emphysema seen on chest computed tomography scan. Ventilation and perfusion scanning was used to identify potential ventilation and perfusion mismatch target areas of lung for plication. RESULTS: The right lung was plicated first in 25 of 32 patients (78%), and upper lobe plications predominated (77%). A mean of 9.3 +/- 0.8 staple firings were used for each unilateral plication procedure. There were no perioperative deaths. Two patients (4%) required axillary thoracotomies to repair air leaks. Mean chest tube duration was 6.3 +/- 0.5 days. Median hospital stay was 7 days (range 3-15). An Intensive Care Unit stay was required following 8 procedures (17%). Postoperative morbidity occurred in 18 (39%) of 46 procedures, including 5 cases of atrial fibrillation and 4 persistent (> 7 days) air leaks. A minimum 2 month followup was available for 22 patients (32 of 46 procedures), demonstrating a clear chest x-ray with significant improvement in ipsilateral diaphragmatic contour. Twelve patients had unilateral reduction, and 10 patients had bilateral reduction in either a staged (n = 7) or sequential at one operation (n = 3) fashion. Twenty-five (78%) of 32 procedures were associated with improved pulmonary function, with a mean increase in FEV1, in patients in this subgroup of procedures, of 43 +/- 7% for each ipsilateral plication at a mean followup of 3.8 +/- 0.5 months. For the entire group of 32 procedures, the mean improvement in measured FEV1 was 29 +/- 7%. Supplemental oxygen requirement was significantly reduced in 9 of 16 patients following plication. CONCLUSION: These data suggest that minimally invasive surgical techniques coupled with a no-cut lung plication can achieve significant lung volume reduction with favorable postoperative morbidity and mortality. Lung plication appears to hold promise as an alternative technique of LVRS. PMID- 9208958 TI - Staging laparoscopy with laparoscopic ultrasonography: optimizing resectability in hepatobiliary and pancreatic malignancy. AB - BACKGROUND: Open laparotomy has traditionally been required to stage hepatobiliary and pancreatic (HBP) cancers accurately. For unresectable patients, costs and morbidity have been high. Today, laparoscopy alone or combined with laparoscopic ultrasonography (LUS) is being examined for its value in defining the extent of malignancy. STUDY DESIGN: We have analyzed the effect of routine implementation of this new staging technique in our HBP center. Staging laparoscopy (SL) with LUS was performed in 50 consecutive patients with HBP malignancies. All patients were considered to have resectable tumors as determined by traditional preoperative staging modalities. Primary tumors were located in the liver (n = 7), biliary tract (n = 11), or pancreas (n = 32). An average of 2.7 preoperative studies per patient were performed prior to SL-LUS. RESULTS: Staging laparoscopy with laparoscopic ultrasonography predicted resectable tumors in 28 patients (56%). At laparotomy, 26 of 28 were actually resectable: the false-negative rate was 4%. Staging laparoscopy with laparoscopic ultrasonography indicated unresectability in 22 patients (44%). Staging laparoscopy alone demonstrated previously unrecognized occult metastases in 11 patients (22%). In 11 other patients (22%) in whom SL alone was negative, LUS established unresectability from vascular invasion (n = 5), lymph node metastases (n = 5), or intraparenchymal hepatic tumor (n = 1). All cases of unresectability due to vascular invasion were validated by laparotomy. Five of six lymph node or hepatic metastases were proved histologically by LUS-guided needle biopsy rather than laparotomy. CONCLUSIONS: Unnecessary laparotomy can be safely avoided by SL LUS in many patients with HPB malignancies, reducing costs and morbidity. PMID- 9208959 TI - Common bile duct exploration and laparoscopic cholecystectomy: role of intraoperative ultrasonography. AB - BACKGROUND: In October 1993, to detect associated common bile duct (CBD) stones, we started an evaluation program of patients with symptomatic cholelithiasis who were candidates for laparoscopic cholecystectomy. STUDY DESIGN: We used a standard preoperative algorithm and a laparoscopic ultrasonographic (LUS) examination. Preoperative endoscopic retrograde cholangiopancreatography (ERCP) was reserved for high-risk patients for CBD stones. Laparoscopic ultrasonographic examination during cholecystectomy was routinely performed to identify stones unsuspected preoperatively. Two-hundred-sixteen patients with symptomatic cholelithiasis were included in the study; 177 patients (82%) were at low risk for choledocholithiasis and 39 patients (18%) were at high risk and had preoperative ERCP. In 17 patients (43.5%) CBD stones were found, and in 16 patients (41%) they were removed by endoscopic sphincterotomy. RESULTS: In all patients, the main intra- and extrahepatic ducts were well documented by LUS, but in eight cases the distal tract of the CBD was not well-visualized. In eight patients, small stones were found in the CBD. A subsequent peroperative cholangiography or CBD exploration confirmed the diagnosis. In one patient, both LUS and cholangiography suspected a small stone; the CBD exploration did not confirm it (false positive). In two patients a small stone in the CBD was found during the followup period (two false negatives). An endoscopic sphincterotomy solved the problem. CONCLUSIONS: Laparoscopic ultrasonographic examination may be a real alternative to cholangiography during laparoscopic cholecystectomy: this may be reserved for selected instances on the basis of LUS findings. On the other hand, considerable ultrasonographic experience is required for LUS to be performed successfully. PMID- 9208960 TI - Laparoscopic or open splenectomy for hematologic disease: which approach is superior? AB - BACKGROUND: This study was undertaken to compare safety, outcome, and costs of laparoscopic (LS) and open splenectomy (OS) for a variety of hematologic diseases. STUDY DESIGN: The records of 137 patients who underwent splenectomy (63 LS and 74 OS) at a large private teaching hospital between March 1991 and April 1996 were reviewed retrospectively. Diagnosis, age, gender, operative time, blood loss, splenic weight, time to resumption of oral diet, postoperative hospital stay, morbidity, mortality, and costs (direct and operative) were analyzed by multivariate statistical analysis. RESULTS: Laparoscopic splenectomy patients had significantly shorter hospitalization and time to resumption of an oral diet (p < 0.01); although operative costs were higher, total direct costs were not. Idiopathic thrombocytopenic purpura patients had earlier resumption of an oral diet after LS, shorter postoperative stay, and comparable OR time. Five patients (7%) were converted, with outcomes similar to OS except for greater operative time and cost. Grade II complications occurred in three LS and four OS patients; Grade III in three OS patients; and Grade IV in two OS patients. There were two major complications of LS and eight of OS, with two deaths. Multivariate analysis showed that operative time and time to resumption of oral intake were significantly related to age, diagnosis, operative technique, and splenic weight. Duration of postoperative hospitalization was related to operative technique, splenic weight, and major complications. Costs (direct and operative) were related to age, splenic weight, and major complications, but not to operative technique. CONCLUSIONS: LS results are influenced by splenic weight, disease, and age. Splenic weight appears to be the crucial determinant of operative time and length of hospitalization. LS is a superior treatment for patients with idiopathic thrombocytopenic purpura and patients with small spleens. PMID- 9208961 TI - Preoperative carcinoembryonic antigen predicts outcomes in node-negative colon cancer patients: a multivariate analysis of 572 patients. AB - BACKGROUND: Although prospective trials have demonstrated that postoperative chemotherapy for node-positive colon cancer patients provides survival benefit, no improvement in survival has been documented for node-negative colon cancer patients. There are, however, a subset of node-negative patients that go on to die of their disease. We hypothesize that this subset of node-negative patients may benefit from postoperative chemotherapy. We analyzed a large cohort of node negative colon cancer patients from a single institution to determine prognostic factors that predict which patients with node-negative colon cancer might experience recurrence and can benefit from postoperative chemotherapy. STUDY DESIGN: A review of the prospective database for colorectal cancer at Memorial Sloan-Kettering Cancer Center (MSKCC) between 1985 and 1993 identified 572 patients who underwent curative resection for node-negative colon cancer (T(1,2,3,4)N0M0). Demographic, serum, and pathologic factors were analyzed for prognostic significance. Survival was calculated by the method of Kaplan-Meier and compared by log rank test. Multivariate analysis was calculated by the Cox proportional hazard model. RESULTS: Median follow-up was 35 months. Factors predictive of survival by univariate analysis include tumor stage, overall stage, and preoperative serum carcinoembryonic antigen (CEA) elevation. By multivariate analysis, overall stage and preoperative serum CEA level predicted survival. CONCLUSIONS: Routine histologic and demographic factors do not predict outcome in node-negative colon cancer patients. Preoperative CEA and overall stage predict survival by multivariate analysis. Preoperative CEA elevation in node-negative patients identifies a group of patients that has a poor prognosis and defines a subset of patients who may benefit from postoperative chemotherapy. PMID- 9208962 TI - The miniscule benefit of serial carcinoembryonic antigen monitoring after effective curative treatment for primary colorectal cancer. AB - BACKGROUND: Serial carcinoembryonic antigen (CEA) levels have been recommended to detect asymptomatic recurrent colorectal cancer and to facilitate curative additional therapy. This study was designed to investigate the outcome of additional treatment for recurrent cancer in patients undergoing primary colorectal cancer treatment in a specialty center and subsequent relapsing with an elevation in serum CEA. STUDY DESIGN: Patients treated for their primary cancers at our institution whose followup included CEA monitoring and whose cancers subsequently recurred, were analyzed from a prospective database of almost 1,900 patients. CEA levels of > or = 5 ng/mL were considered elevated for purposes of treatment results. One hundred sixty-three patients were suitable for analysis. Median followup before and after recurrence was 14 months and 16 months, respectively. RESULTS: Fifty patients were able to undergo complete resection of their recurrence, and 26 of these patients are without evidence of recurrence at last followup. Two-thirds of recurrences were associated with an elevation of CEA; this elevation at recurrence was associated with decreased survival (p < 0.05, Kaplan Meier). Of the 109 patients with an elevation of CEA at recurrence, complete re-resection was accomplished in 26 patients. Of these, half remain cancer free. Of those with a normal CEA at recurrence, complete re resection was feasible in 24 patients. CONCLUSIONS: Only 17% of patients with recurrent colorectal cancer undergoing potentially curative reresection have an elevated CEA. If we use the denominator of our patient population using an estimated relapse rate of 25-50%, the overall likelihood of CEA-directed curative re-resection confirms early estimates of less than a 5% survival advantage. PMID- 9208963 TI - Metastatic gastric lymph node rate is a significant prognostic factor for resectable stage IV stomach cancer. AB - BACKGROUND: Stage IV gastric carcinoma is rarely curatively resected and almost invariably carries a poor prognosis. Several clinicopathologic factors are involved, but lymphatic spread of the cancer may significantly affect survival. STUDY DESIGN: A retrospective study was designed to evaluate whether the nodal metastatic rate (number of lymph node metastases/number of resected lymph nodes) is a parameter of lymphatic spread and could provide a significant prognostic factor. Several prognostic factors were assessed by multivariate analysis in 153 stage IV gastric carcinoma patients with histopathologic data on nodal metastasis. RESULTS: A significant difference in survival was observed in the stage IV cancer patients with total nodal or gastric nodal metastatic rates < 50% versus those with rates > 50%. Multivariate analysis revealed that a total nodal or gastric nodal metastatic rate > 50% was a highly significant prognostic factor. The gastric nodal metastatic rate can be used in patients who do not undergo an extended lymphadenectomy. CONCLUSIONS: Lymphatic spread of gastric carcinoma expressed in terms of the total nodal or gastric nodal metastatic rate is a significant prognostic factor. The latter can be calculated without pathologic data derived from extended lymphadenectomy, and so it is universally applicable. PMID- 9208964 TI - Appraisal of the order of revascularization in human liver grafting: a controlled study. AB - BACKGROUND: By current convention, the liver graft is revascularized, first with portal blood flow, and thereafter with arterial blood flow. Although experimental studies showed no detrimental effects of primary arterialization, this order of revascularization has not been investigated in clinical transplants. STUDY DESIGN: Twenty-nine patients were included in our controlled study to investigate and compare, by means of a technical procedure that permits either initial arterial revascularization (IAR) or initial portal revascularization (IPR), the effects of graft revascularization by IAR and by IPR in clinical transplants. RESULTS: Patients were equally divided in the IAR group (n = 15) and the IPR group (n = 14), and were homogeneous in terms of recipients and graft characteristics. Graft reperfusion was uniform and diffuse in all grafts with IAR versus 10 (71%) with IPR (p < 0.05). After reperfusion, the time taken for completion of the procedure was shorter in the IAR group (159 +/- 28 versus 242 +/- 39 minutes) (p < 0.01). Both mean blood transfusions and antifibrinolytic requirements were lower in the IAR group: 5.4 +/- 1.8 versus 7.6 +/- 3.5 packed red cell units, and 13% versus 50%, respectively (p < 0.05). Postoperative ASAT level, clotting factor V level, and bile flow were not different between the two groups. Early postoperative vascular or biliary complications did not occur. During a mean follow-up of 16 months (range, 7-20), one hepatic artery thrombosis occurred in the IPR group, and one anastomotic biliary stricture occurred in each group. CONCLUSION: Under adequate portal decompression, LAR is a safe option and results in better graft reperfusion, shorter post revascularization phase, and reduced transfusion and antifibrinolytic requirements. PMID- 9208966 TI - Blunt carotid artery injuries. AB - BACKGROUND: Blunt carotid artery trauma remains a rare but potentially devastating injury. Early detection and treatment remain the goals of management. Our objective was to identify patients sustaining blunt carotid injuries at a regional trauma center and report on the incidence, demographics, diagnostic workup, management, and outcome. STUDY DESIGN: A retrospective chart review was performed of patients sustaining blunt carotid artery injury between 1990 and 1996. RESULTS: Twenty patients were identified during the 7-year period. All patients suffered blunt trauma, with motor vehicle accidents being the most common mechanism, and the internal carotid the most frequently injured vessel. Associated injuries were present in all patients, with head (65%) or chest (65%) injuries being the most common. The combination of head and chest trauma (45%) was found to be associated with a 14-fold increase in the likelihood of carotid injury. Cerebral angiography was diagnostic in all patients and the majority were treated nonoperatively with anticoagulation. Twenty percent of patients were discharged with a normal neurologic exam, while 45% left with a significant neurologic deficit. Overall mortality was 5%. CONCLUSIONS: Blunt carotid injuries are rare but are associated with significant morbidity and mortality. The combination of craniofacial and chest wounds should raise the index of suspicion for blunt carotid injury. Anticoagulation was associated with the least morbidity. PMID- 9208965 TI - A prospective comparison of two expanded polytetrafluoroethylene grafts for linear forearm hemodialysis access: does the manufacturer matter? AB - BACKGROUND: The function and patency of standard 6-mm Goretex (W.L. Gore and Associates, Flagstaff, AZ) and Impra (Impra, Inc., Tempe, AZ) expanded polytetrafluoroethylene (e-PTFE) grafts for hemodialysis as radial-antecubital linear arteriovenous fistulae for dialysis are compared. STUDY DESIGN: A randomized clinical trial was conducted in two community dialysis centers and in one hospital-based center serviced by one vascular surgical practice, that performed the access surgery. Selection of linear forearm access, as opposed to other hemodialysis graft configurations, was at the discretion of the surgeon. Candidates for linear grafts had palpable radial pulses with a normal Allen test and normal digital Doppler flow in the hand. Linear grafts were placed using end to-side anastomoses to the artery and vein, and the graft type was determined by randomization. Primary patency was determined by first episode of thrombosis, first revision, or angioplasty of the graft. Secondary patency after thrombectomy, revision, or angioplasty was determined when the graft was no longer clinically usable, and a new graft needed to be placed as a parallel conduit in the forearm or in another site. Statistical analysis was by actuarial life-table methods. RESULTS: There were 131 linear forearm grafts in 117 patients. The Impra and Goretex groups were equally matched for gender and major risk factors, except for smoking, which was more common in the Goretex group. Minimum followup was 24 months. Life table primary patencies at 1 year (Impra 43%, Goretex 47%) and at 2 years (Impra 30%, Goretex 26%) were not statistically different (p = 0.78); secondary patency was also equal at 1 year (Impra 49%, Goretex 69%) and at 2 years (Impra 33%, Goretex 41%) (p = 0.15). Discontinuance of use of a patent graft, complications, episodes of thrombosis, and the need to replace the original graft occurred in the two groups without a statistically significant difference. CONCLUSIONS: In the linear forearm position from the radial artery to an antecubital vein, there is no difference in the performance of 6-mm standard e-PTFE grafts on the basis of manufacturer, whether Goretex or Impra. On the basis of performance, linear forearm dialysis grafts are an acceptable method for hemodialysis access. PMID- 9208967 TI - Stomal ulcer after gastric restrictive operations. PMID- 9208968 TI - Mesh-foil laparostomy. PMID- 9208969 TI - Useful stapling techniques in liver surgery. PMID- 9208970 TI - Specimen adequacy in breast fine-needle aspiration biopsy. PMID- 9208971 TI - Mentor-based management training. PMID- 9208972 TI - Cytokeratins, CEA, and mucin histochemistry in the diagnosis and characterization of extramammary Paget's disease. AB - To identify a sensitive marker for extramammary Paget's disease and to identify histochemical and immunohistochemical features that suggest occult pelvic cancer in patients with extramammary Paget's disease, we retrieved all cases between 1983 and 1992 with a Standardized Nomenclature of Medicine code of extramammary Paget's disease in the Vanderbilt University Medical Center (Nashville, Tenn) surgical pathology archives. All were stained for alcian blue/dPAS (periodic acid Schiff), mucicarmine, AE1/AE3, cytokeratin (CAM 5.2), cytokeratin (CK) 7, CK 20, carcinoembryonic antigen (CEA), orthokeratin, prostate-specific antigen, and S 100. Sixteen cases (2 men, 14 women) were retrieved. Two had pelvic malignancies: one rectal adenocarcinoma and one transitional carcinoma. Only CK7 marked all cases. Mucins were sensitive but focal, a potential problem in small biopsy specimens. The transitional tumor had a unique staining profile (CEA- and mucin negative). CK20 strongly marked Paget cells associated with rectal cancer; its presence suggests a large bowel lesion but is not specific. No case expressed prostate-specific antigen; its presence in a man suggests prostatic carcinoma. PMID- 9208973 TI - In search of specimen adequacy in fine-needle aspirates of nonpalpable breast lesions. AB - Pathology-related medical malpractice claims frequently concern fine-needle aspirations (FNAs) of breast lesions, and diagnostic errors have been attributed in part to the inadequacy of the specimens. Cytologic criteria for adequate FNA specimens, specifically in cases without malignancy, have not been clearly defined. From January 1988, to August 1995, 669 ultrasonographic-guided FNAs of nonpalpable, solid breast lesions with subsequent histologic examination were performed at our institution. From these, 54 cases with cytologic diagnoses of insufficient or nonspecific benign findings were identified. All aspirates were reviewed, and the number and size of the epithelial cell groups were quantitated in each case. By using criteria for adequate aspirates of palpable breast lesions (six or more epithelial cell groups per case with a minimum of 5-10 cells per group), 23 of the 54 aspirates were deemed inadequate and 31 adequate. Eleven (48%) of the inadequate aspirates and 17 (55%) of the adequate aspirates were from histologically confirmed carcinomas (ductal carcinoma in situ, 6; invasive carcinoma, 22, of which 12 were ductal, 7, lobular, and 3, mixed ductal and lobular). For the mammographic diagnoses "probably benign," "indeterminate," and "suggestive of malignancy or malignant," the probability of malignancy in aspirates of adequate cellularity (eg, > 6 epithelial groups) was 9%, 40%, and 93%, respectively. These findings indicate that a significant proportion of breast aspirates still may yield false-negative results despite adequate to high cellularity. Although a definition of adequacy based on cellularity is useful in reducing false-negative results, cellularity alone cannot be relied on in the management of nonpalpable lesions. For mammographic findings that are indeterminate or suggestive of malignancy or malignant, nonspecific FNA findings should be followed by core or excisional biopsy to exclude carcinoma. PMID- 9208974 TI - Leiomyosarcoma of the female breast: report of two new cases and a review of the literature. AB - We present the clinical, light microscopic, and immunohistochemical features of two new cases of leiomyosarcoma of the female breast. Both the patients were old (83 and 86 years) and were referred with a history of a long-standing breast lump. The results of the physical examination and the preoperative radiologic investigations suggested a phyllodes tumor. The patients were treated with mastectomy. The tumors measured 6 and 6.5 cm in the largest dimension, respectively, and were composed of fascicles of atypical, actively proliferating spindle cells, often intersecting at right angles. The axillary lymph nodes were free of tumor. Immunohistochemically, the tumor cells were positive for desmin, muscle-specific actin, and vimentin and negative for other markers, including keratins and hormone receptors. Focal rhabdomyoblastic differentiation was noted in one case. Follow up at 1 year is negative for metastases or local recurrences. Our study confirms that leiomyosarcoma of the breast is a locally invasive neoplasm and that it could represent a peculiar anatomic entity among mesenchymal tumors of the breast. PMID- 9208975 TI - Enhanced cellular proliferation and p53 accumulation in gastric mucosa chronically infected with Helicobacter pylori. AB - This study evaluated whether the increased risk of development of gastric carcinoma due to chronic Helicobacter pylori infection could be linked with elevated cell proliferative activity and expression of p53 and bcl-2. Forty-eight patients undergoing therapy for H pylori-positive gastroduodenal ulcers were separated into not eradicated (NE; n = 23) and eradicated (E; n = 25) groups 6 months after the treatment. Serum pepsinogen (PG) I:II ratios and histologic changes in the gastric corpus and the antrum, assessed according to the modified Sydney System, as well as epithelial cell proliferation (mitosis, Ki67, and proliferating cell nuclear antigen [PCNA]), and expression of oncoproteins (p53 and bcl-2) were examined before and at 3 months and 6 months after treatment for H pylori. Chronic persistent H pylori infection was associated with a low PG I:II ratio, increased inflammation and activity score, and elevated cell proliferation, as evidenced by the Ki67 and PCNA labeling indexes and the mitotic index in the NE group. Scattered accumulation of p53 protein continued to be observed in the NE group after treatment but was significantly decreased in the E group. We conclude that persistent H pylori infection causes gastritis, with epithelial degeneration and regeneration that result in accentuation of epithelial cell proliferation and accumulation of p53 protein, presumably heightening the genetic instability consistent with the development of carcinoma. PMID- 9208976 TI - Round cell liposarcoma with the insertion (12;16)(q13;p11.2p13). AB - Cytogenetic analysis of a short-term culture from a round cell liposarcoma revealed ins(12;16)(q13;p11.2p13) as a sole abnormality in all metaphase cells studied. This chromosome rearrangement, thus far not described in liposarcomas, leads to recombination of bands 12q13 and 16p11.2 and, thus, seems to be the equivalent of t(12;16)(q13;p11), a translocation that is highly specific for the myxoid type of liposarcoma. Our case represents the fourth fully karyotyped round cell liposarcoma that displays a cytogenetic rearrangement of bands 12q13 and 16p11, thus supporting the concept that round cell liposarcoma is related to myxoid liposarcoma and constitutes its poorly differentiated form. The fact that ins(12;16) was the only detectable chromosome aberration suggests that the presence of secondary cytogenetic aberrations is not a prerequisite for the development of a round cell histology in liposarcoma. PMID- 9208977 TI - Sarcomatoid carcinomas of the lung: a clinicopathologic review. AB - Sarcomatoid carcinomas (SC) of the lung are the most common pulmonary neoplasms that exhibit a composition by spindled or pleomorphic tumor cells. As such, many of them may be confused easily with true sarcomas diagnostically unless special immunohistological or ultrastructural analyses are performed. Reactivity is expected for keratin, epithelial membrane antigen, or collagen type IV in the sarcomalike elements in SC, although it may be focal. Electron microscopy often shows the presence of junctional complexes between tumor cells, with or without pericellular basal lamina and cytoplasmic skeins of intermediate filaments. Current terminological preferences are such that several formerly used terms- including "spindle-cell carcinoma," "pulmonary blastoma," "squamous cell carcinoma with pseudosarcomatous stroma," "pseudosarcoma," and "carcinosarcoma"- are now encompassed by the more generic designation of "sarcomatoid carcinoma." The clinical course of patients with this neoplasm is aggressive, with an overall 5-year survival rate approximating 20%. PMID- 9208978 TI - Immunophenotype of Reed-Sternberg and Hodgkin's cells in sequential biopsy specimens of Hodgkin's disease: a paraffin-section immunohistochemical study using the heat-induced epitope retrieval method. AB - Other studies have shown that the immunophenotype of Reed-Sternberg and Hodgkin's (RS-H) cells in Hodgkin's disease commonly changes over time, as shown by examination of multiple biopsy specimens obtained from an individual patient. In this study we analyzed 96 sequential biopsy specimens (>1 month apart) obtained from 44 patients (nodular sclerosis, 34 specimens; mixed cellularity, 5; lymphocyte depletion, 1; unclassified, 4) using fixed, paraffin-embedded sections; heat-induced epitope retrieval (HIER); a panel of antibodies specific for the CD3, CD15, CD20, CD30, CD43, CD45/45RB, and CD79a antigens and Epstein Barr virus latent-membrane protein; and a streptavidin-biotin method. In selected cases in which immunophenotypic changes occurred, studies were repeated using enzyme predigestion instead of HIER. There was no change in the immunophenotype of the RS-H cells in 36 (82%) of 44 patients. In 8 patients (18%), the immunophenotype of the RS-H cells varied in expression of one or two antigens. The antigens that varied were as follows: CD30, 3 patients; CD15, 3 patients; CD20, 1 patient; and CD15 and CD30, 1 patient. We conclude that the immunophenotype of RS-H cells in Hodgkin's disease is relatively stable over time and that CD15 and CD30 are the most common antigens that change. The frequency of immunophenotypic changes, 18%, is substantially lower than that reported previously. One likely explanation for this discrepancy is that we used HIER, a relatively recent innovation in diagnostic immunohistochemistry that has been shown to reduce artifacts attributable to inconsistent fixation and processing. The significance of immunophenotypic variation in eight cases (18%) is uncertain. This phenomenon may represent true biologic changes in RS-H cells. Alternatively, these changes may be attributable to artifacts secondary to inconsistent fixation or processing that HIER cannot overcome. PMID- 9208979 TI - Assessment of clonality in cutaneous lymphoid infiltrates by polymerase chain reaction analysis of immunoglobulin heavy chain gene rearrangement. AB - Determination of the biologic potential of cutaneous lymphoid infiltrates may be difficult by standard histologic or immunohistologic examination. The polymerase chain reaction (PCR) has been used to document clonal rearrangements of the immunoglobulin heavy chain gene in paraffin-embedded fixed tissues. To explore the value of PCR in evaluation of cutaneous lymphoid infiltrates, 93 archival nonmycotic lymphoid lesions (28 small or mixed lymphocytic lymphomas, 15 large cell lymphomas, 40 benign infiltrates, and 10 with atypical features) were analyzed. These cases had been previously immunophenotyped on paraffin sections, and clinical follow-up from 7 to 20 years was gathered. DNA products were generated using a seminested PCR technique, separated by 5% polyacrylamide gel electrophoresis, stained with ethidium bromide, and visualized under UV light. Cases with a 100- to 120-base pair band were scored as positive. Of the 28 small or mixed cell lymphomas, 23 had a B-cell immunophenotype or consisted of a mixture of B and T cells; of these, seven (30%) demonstrated a monoclonal pattern, and three (13%) were indeterminate. Twelve large cell cases were B-cell or mixed; five (42%) of these were positive for a monoclonal band, while four (33%) were indeterminate. None of five T-cell small cell or three T-cell large cell lymphomas demonstrated a monoclonal band. In contrast, 39 of the 40 benign cases were T-cell predominant or mixed lesions. Nevertheless, 18 of these 40 cases on initial testing suggested possible monoclonality. Six were indeterminate, and 12 demonstrated apparent monoclonal bands, of which four were reproducible on repeat testing. No histologic or clinical features of lymphoma were present in 17 of these 18 cases, suggesting that they represent false positive results. Most of the latter lesions showed sparse perivascular infiltrates, with very few B cells. This suggests that amplification of the immunoglobulin heavy chain gene from a small number of lymphocytes may produce a monoclonal band. In summary, PCR may provide adjunct information about clonality in selected lymphoid skin lesions, but is rather insensitive in this setting. Such data must be carefully considered in the context of the histologic, immunohistologic, and clinical findings, particularly when assessing sparse infiltrates, because of the potential for false-positive results. PMID- 9208980 TI - Clinical significance of immature reticulocyte fraction determined by automated reticulocyte counting. AB - The Sysmex R-3000 (TOA Medical Electronics, Kobe, Japan) evaluates maturation of reticulocytes by quantitating the fraction of reticulocytes within low-, middle-, and high-fluorescence intensity regions. We defined the immature reticulocyte fraction (IRF) as the sum of the fraction of high-fluorescence intensity regions plus the fraction of middle-fluorescence intensity regions. Then, we studied the clinical significance of IRF in the evaluation of anemia by comparing the IRF with the absolute reticulocyte count (ARC) and with the reticulocyte production index (RPI) and by reviewing pertinent clinical information about the patients. In the study, 132 specimens from 102 patients undergoing evaluation of anemia were analyzed. By using simple regression analysis, our results showed that the IRF has a weak but significantly positive correlation with ARC and with RPI, indicating that IRF is an additional useful parameter to evaluate the erythropoietic activity in anemia. Interpretation by integrating IRF and reticulocyte enumeration (ARC and RPI) provided useful information for further subclassification of anemia. Increased IRF (IRF > or = 0.23) and increased ARC generally indicated an adequate erythroid response to anemia. All but three specimens with an IRF less than 0.23 showed an RPI of 2 or less. These specimens were from patients with underlying diseases known to lead to decreased erythropoietic activity, predominantly chronic renal insufficiency. Specimens with a subnormal or normal ARC (with a corresponding RPI < or = 2) but with an IRF of more than 0.23 were from patients with various underlying conditions, including acute infection, iron deficiency anemia, human immunodeficiency virus infection, sickle disease with crisis, pregnancy, and myelodysplastic syndrome. Our results indicate that an IRF of 0.23 or less in patients with anemia reflects bone marrow that is nonresponsive or underresponsive to the anemia. Patients with an increased IRF (IRF > or = 0.23) may require further examination to clarify the cause of the anemia. PMID- 9208981 TI - Comparison of an activated partial thromboplastin time with a Russell viper venom time test in screening for factor V(Leiden) (FVR506Q). AB - Factor V(Leiden) is the most common abnormality detected in patients examined because of hereditary thrombophilia. The most widely used clot-based screening test is based on the activated partial thromboplastin (aPTT) time. This test has a low sensitivity. A comparison of the aPTT-based test with a Russell viper venom time test (RVVT) was performed in matched samples. All samples were analyzed by polymerase chain reaction (PCR) for the factor V(Leiden) defect. We studied 139 samples, of which 109 were PCR-negative; 30 were PCR-positive. Using the manufacturer's suggested threshold ratio of 2, the aPTT test showed a sensitivity of 0.43, a specificity of 0.86, and a positive predictive value (PPV) of 0.97. The RVVT test had a sensitivity of 1.0, a specificity of 0.95, and a PPV of 0.91. Segregation of a subpopulation of this study population into ABO group O vs non group O showed an effect of ABO group on the aPTT test but not on the RVVT test, consistent with an influence of factor VIII clotting (factor VIII:C) on the aPTT test. The RVVT test seems superior to the unmodified aPTT test as a screening test for factor V(Leiden). PMID- 9208982 TI - Heparin-induced platelet aggregation vs platelet factor 4 enzyme-linked immunosorbent assay in the diagnosis of heparin-induced thrombocytopenia thrombosis. AB - Thrombosis occurs in an unpredictable subset of patients with heparin-induced thrombocytopenia (HIT). The diagnosis of HIT requires clinical suspicion and laboratory confirmation. Although the "gold-standard" diagnostic test is considered to be the serotonin release assay (SRA), most laboratories use heparin induced platelet aggregation (HIPA), which is highly specific but reported to be less sensitive than the SRA. Recently, the heparin-platelet factor 4 (PF4) enzyme linked immunosorbent assay (ELISA) has been reported to have comparable sensitivity to the SRA. We compared the HIPA and PF4 ELISA in serum samples from 146 patients examined for HIT and assessed whether either test predicted thrombotic risk. Results for 81 patients were positive for HIPA, PF4 ELISA, or both. Of these, 91% were HIPA-positive, while only 60% were PF4 ELISA-positive. Clinical information was available on 63 patients, 17 of whom had thrombotic events (10 venous, 6 arterial, and 1 both). Neither the HIPA nor the PF4 ELISA predicted thrombotic risk, but the HIPA proved to be a more sensitive test for laboratory confirmation. PMID- 9208983 TI - Long-term French experience in INR standardization by a procedure using plasma calibrants. AB - The International Normalized Ratio (INR) has not lowered the interlaboratory differences in prothrombin time (PT) values to the extent expected, mainly because of the instrument-dependency of the International Sensitivity Index (ISI) and other factors (eg, accurate determination of the ISI, the normal value used in the PT ratio). The procedure (PPC) using plasma calibrants (reference lyophilized plasmas with assigned activity) has been evaluated since 1977 in nine French national external quality assessment surveys (NEQAS) involving approximately 4,000 laboratories and numerous local thromboplastin technique combinations. The PPC was compared with the conventional procedure (using the manufacturer's ISI), and the efficiency of antivitamin K-calibrated (AK Cal) plasmas from patients receiving oral anticoagulants vs artificially depleted plasma calibrants was also evaluated. The PPC efficiently standardized PTs with AK Cal plasmas, reducing interlaboratory variability (eg, coefficient of variation, 12% to 6% for survey 92 D) and reagent-instrument effects. However, AK Cal plasmas have drawbacks, such as limited supply, cost, and batch-to-batch variability. The artificially depleted plasma calibrants were less efficient, but usable if carefully prepared. The value of this simple procedure is that local practices are considered in the determination of PT, thus correcting for coagulometer effects and avoiding use of the manufacturer's ISI and need for a normal control plasma. These large-scale French surveys have demonstrated the validity of PPC through 15 years of experience and have shown that it offers the best compromise available in PT standardization. PMID- 9208984 TI - A leadership-management training curriculum for pathology residents. AB - Leadership and management skills are increasingly critical to the success of pathologists in and outside of academic centers. We describe a leadership and management curriculum in a combined anatomic and clinical pathology residency program. The mentor-based approach incorporates leadership and management skills throughout the residency with a dedicated 2-month rotation during the final year of training. This 2-month rotation is led by a senior faculty pathologist mentor and includes active participation in management activities, small group discussions, reading, and a management process project. The topics for the small group discussions, reading list, mentor-based activities, and completed management process projects are described. Outcomes and evaluations are reported for residents who have completed this curriculum in leadership and management. PMID- 9208985 TI - Survey of management training in United States and Canadian pathology residency programs. AB - One hundred seventy-six United States and Canadian pathology residency programs were surveyed to assess the status of management training for residents. Specific questions in the survey concerned approaches to management, instructional methods used, topics covered, and length of management rotations. Eighty-four programs (48%) responded to the survey. This group represented a cross section of all programs as to location, size, and type of program. Of the 84 programs, 81 (96%) offered management training, while 54 programs (64%) offered a management course. Management training usually occurred in combination with other rotations, most frequently clinical pathology subspecialties. A dedicated management rotation was reported in 19 programs (23%). The most common method of instruction was apprenticeship/mentor followed by didactic lecture. A majority of programs used multiple instructional methods. The five most frequently covered topics were budgets, personnel issues, quality assurance, utilization, and instrument evaluation, but a wide variety of topics were included in management curricula surveyed. There was an emphasis on clinical laboratory administration. Nonpathologist instructors, such as medical technologists, hospital administrators, and business executives, were used for formal didactic exercises, while teaching by pathologists occurred predominantly by mentoring during clinical pathology rotations. PMID- 9208986 TI - Papanicolaou smear sensitivity for adenocarcinoma in situ. PMID- 9208987 TI - Hyperhomocysteinemia. PMID- 9208988 TI - Hyperhomocysteinemia. PMID- 9208989 TI - Distribution of vasopressin and vasoactive intestinal polypeptide (VIP) fibers in the human hypothalamus with special emphasis on suprachiasmatic nucleus efferent projections. AB - The human suprachiasmatic nucleus (SCN) is located in the basal part of the anterior hypothalamus and is considered as the biological clock that generates circadian rhythms and synchronizes the daily activity pattern with the environmental light-dark cycle. However, the mechanisms and pathways by which the SCN transmits its information to the other brain areas are unknown. Therefore, in the present study, we investigated the efferent projections of the SCN by the immunocytochemical staining of two major peptidergic SCN neurotransmitters: vasopressin (VP) and vasoactive intestinal polypeptide (VIP). It confirmed that these peptides are present in different subdivisions of the SCN. The results of this investigation show that VP and VIP fibers arising from the SCN were detected to branch extensively and hence seem to innervate the SCN itself and the central and medial part of the anteroventral hypothalamic area (AVH), the area below the paraventricular nucleus (sub-PVN), the ventral part of the paraventricular nucleus (PVN), and the dorsomedial nucleus of the hypothalamus (DMH). There appeared to be substantial congruity between the presumptive human SCN projections and those as observed by tracing in rat or hamster. Regarding the anatomical organization of the human SCN projections, the main projection areas appeared to be the AVH, the sub-PVN, the ventral part of the PVN, and the DMH. The observation that VIP and in particular VP fibers pass between the SCN and the PVN suggests that the human SCN and the PVN may have a direct anatomical connection. In addition, VP and VIP fibers were detected in several other hypothalamic areas that are not known to have clear direct connections to the SCN. The possible origin of these VP and VIP fibers is discussed. PMID- 9208990 TI - A compartment-based, asymmetric representation of the retina in an induced projection to the olfactory cortex. AB - Displacing the optic nerve into the telencephalon in adult Rana pipiens induces a projection to olfactory cortex. We have examined the topographic organization of this projection anatomically by injecting a mixture of biotin dextran (BDA) with 3H-amino acids into the affected eye immediately after making cuts across defined sectors of the nerve fiber layer to trace the complementary patterns of anterograde migration of BDA and 3H label in the cut and intact retinal axons, respectively. Fibers from the temporal side of the optic disc terminated in an oblique band along the posterior two-thirds or more of the ectopic projection field. In contrast, fibers arising in the nasal retina terminated in a parallel strip occupying the anterior one-third or less of the field. Varying the location of the cuts within each hemiretina did not reveal any further organization along the nasotemporal or dorsoventral axes of the retina. The retinal location of the cells involved in this projection was further studied with injections of wheat germ agglutinin conjugated to horseradish peroxidase into the olfactory cortex. Ganglion cells labeled by retrograde transport were found throughout the retina, but they were much more numerous on the temporal side, having a mean spatial density 3.7-7.4 times greater in the temporal hemiretina, whereas the overall ganglion cell density (labeled plus unlabeled) was roughly the same in the two halves of the retina. These data provide an example of a permanent projection in which the overall representation of the retina, though nontopological, is polarized in one axis (nasotemporal) and, therefore, compartmentally organized. PMID- 9208991 TI - Anatomical organization of efferent neurons innervating various regions of the rabbit masseter muscle. AB - Previous studies have shown that the masseter muscle is supplied by motoneurons located in the anterodorsal region of the trigeminal motor nucleus and by an additional group of efferent neurons located in cell group k. The present experiments were performed on nine rabbits and were designed to establish the locations of neurons innervating the different regions of this muscle. Retrograde labeling with two fluorescent tracers (FluoroGold and Fast Blue) was applied to the central ends of cut branches of the masseter nerve. Serial coronal sections of the brainstem were viewed with fluorescence microscopy. The labeled cells were counted in all animals, and three-dimensional reconstructions of their distribution were made in five cases. In each successful experiment, labeled neurons were seen in the anterodorsal region of the trigeminal motor nucleus and in the two dorsal cell columns of cell group k (k1 and k3). Within-animal comparisons of the median position of populations innervating two distinct muscle regions in five rabbits showed that there were no significant differences in either the dorsoventral or rostrocaudal axes. However, in each case, there was a small but significant difference (83-173 microm) in the mediolateral axis within the motor nucleus but not within cell group k. Even in this axis, there was a 94 99% overlap of the two populations. Comparisons of the neuronal cross-sectional area showed that the deep regions were innervated by a larger proportion of small neurons from both nuclei than were the superficial and intermediate regions. Our results suggest that there is no simple topographical arrangement of motoneurons that corresponds to the peripheral pattern of nerve supply to the different regions of the masseter muscle. PMID- 9208992 TI - Further evidence for the involvement of the spinoparabrachial pathway in nociceptive processes: a c-Fos study in the rat. AB - We have analysed in briefly anaesthetised rats (1% halothane for 18 minutes) the effects of innocuous and noxious heat, applied to the hindpaw, on evoked c-Fos immunoreactivity at the levels of the parabrachial area (PB), spinal cord, and nucleus of the solitary tract (NTS). After anaesthesia recovery, animals were left to move freely for 2 hours. At the spinal level, c-Fos was expressed primarily in the ipsilateral superficial laminae, increasing with the applied temperatures in a dependent manner in the noxious range (correlation coefficient r = 0.954, n = 20). At the NTS level, no noxiously evoked c-Fos expression was observed. At the PB level, c-Fos was expressed preferentially contralaterally, increasing with the applied temperatures in a dependent manner in the noxious range (r = 0.971, n = 25). The maximum expression was observed in the outer portion of the external lateral, the lateral crescent, and the superior lateral subnuclei around the pontomesencephalic junction. This was congruent with the densest supraspinal projection of lamina I neurones of the dorsal horn. Labelling in the PB area was highly correlated (r = 0.936, n = 20) with labelling in the superficial laminae. We conclude that, under our experimental procedures, noxious heat-induced c-Fos expression at the PB level depends on the intensity of the noxious stimulation. These data further support the relevance of the recently described spino-PB pain pathway. Because of their location, the Fos immunoreactive neurones observed in the pontine and the mesencephalic divisions of the PB area were likely PB-amygdaloid and PB-hypothalamic nociceptive neurones, respectively. PMID- 9208993 TI - Size and distribution of retinal ganglion cells in the St. Kitts green monkey (Cercopithecus aethiops sabeus). AB - The topographical distribution of density and the soma size of retinal ganglion cells (RGCs) were studied in the St. Kitts green monkey (Cercopithecus aethiops sabeus). The total number of RGCs, estimated from light microscopic analysis of wholemounted and of transversely sectioned retinae, ranged between 1,183,721 and 1,273,715 (mean 1,228,646). These estimates are comparable to the number of optic nerve fibres (1,220,000) estimated from semithin sections. The topographic distribution of RGCs shows a strong centroperipheral gradient. The soma size distribution of RGCs in Nissl-stained flatmounts falls within a range of between 5.7 microm and 22.9 microm and is comparable to other primate species. Somata of RGCs were found to be generally smaller within the fovea than in peripheral regions. Ganglion cells, as reported for other diurnal primates, are nonuniformly distributed with a slight nasotemporal elongation of isodensity contours, and they exhibit nasotemporal asymmetry in the frequency distribution of soma size. The topography of the RGC distribution of this semiarboreal, ground-dwelling monkey is similar to what has been found in other diurnal Old World species. PMID- 9208994 TI - Glycine immunoreactivity in the lateral nucleus of the trapezoid body of the cat. AB - The central auditory system contains several predominantly glycine-immunoreactive nuclei, and one of these, the lateral nucleus of the trapezoid body, contains cell bodies exhibiting a spectrum of labeling intensity. By using post-embedding glycine immunocytochemistry on thin sections, and toluidine blue staining of adjacent sections, we established that darkly glycine-immunoreactive neurons constituted a distinct morphological class and form one of three subnuclei of the lateral nucleus of the trapezoid body, called the posteroventral subnucleus. These neurons resemble, in both labeling intensity and cell body morphology, the principal cells of the medial nucleus of the trapezoid body. The other two subnuclei of the lateral nucleus of the trapezoid body, its main and hilus subnuclei, contained predominantly glycine-immunoreactive and glycine immunonegative neurons, respectively. Glycine immunoreactivity was compared with gamma-aminobutyric acid (GABA) immunoreactivity in order to identify other organizational features of the lateral nucleus of the trapezoid body. Cell bodies that displayed either dark glycine-immunoreactivity or which were glycine immunonegative were GABA-immunonegative. Cell bodies that displayed GABA immunoreactivity were preferentially located in the main subnucleus. Patterns of distribution of axosomatic innervation in the lateral nucleus of the trapezoid body were revealed in which glycine-immunoreactive puncta were (1) more numerous than GABA-immunoreactive puncta on glycine-immunonegative cell bodies and (2) equal to or less numerous than GABA-immunoreactive puncta on glycine immunoreactive cell bodies. The characteristics of neural circuitry revealed by glycine and GABA immunoreactivity in the lateral nucleus of the trapezoid body may be generalizable to other populations of neurons of the superior olivary complex and to other regions of the central nervous system containing glycinergic neurons, such as the retina. PMID- 9208995 TI - Septal complex of the telencephalon of the lizard Podarcis hispanica. II. Afferent connections. AB - The afferent connections to the septal complex were studied in the lizard Podarcis hispanica (Lacertidae) by means of a combination of retrograde and anterograde tracing. The results of these experiments allow us to classify the septal nuclei into three main divisions. The central septal division (anterior, lateral, dorsolateral, ventrolateral, and medial septal nuclei plus the nucleus of the posterior pallial commissure) receives a massive, topographically organized, cortical projection (medial, dorsal, and ventral areas) and widespread afferents from the tuberomammillary hypothalamus and the basal telencephalon. Moreover, it receives discrete projections from the dorsomedial anterior thalamus, the ventral tegmentum, the midbrain raphe, and the locus coeruleus. The ventromedial septal division (ventromedial septal nucleus) receives a massive projection from the anterior hypothalamus, dense serotonergic innervation, and a faint amygdalohypothalamic projection, but it is devoid of direct cortical input. The midline septal division (nucleus septalis impar and dorsal septal nucleus) receives a nontopographic cortical projection (dorsomedial and dorsal cortices) and afferents from the preoptic hypothalamus, the dorsomedial anterior thalamus, the midbrain central gray, and the reptilian A8 nucleus/substantia nigra. Our results indicate that the cortex provides a physiologically complex, massive input to the septum that terminates over the whole dendritic tree of septal cells. In contrast, most of the ascending afferents make axosomatic contacts by means of pericellular nests. The chemical nature of the main septal afferents and the comparative implications of the available hodological data on the organization of the septal complex of tetrapod vertebrates are discussed. PMID- 9208996 TI - Tracer coupling pattern of amacrine and ganglion cells in the rabbit retina. AB - We examined the tracer coupling pattern of more than 15 morphological types of amacrine and ganglion cells in the rabbit retina. Individual cells were injected intracellularly with the biotinylated tracer Neurobiotin, which was then allowed to diffuse across gap junctions to label neighboring neurons. We found that homologous and/or heterologous tracer coupling was common for most proximal neurons. In fact, the starburst amacrine cell was the only amacrine cell type that showed no evidence of coupling. The remaining types of amacrine cell were coupled exclusively to other amacrines, either homologously or, more often, through a combination of homologous and heterologous junctions. In only one case did we visualize labeled ganglion cells following injection of Neurobiotin into an amacrine cell. In contrast, injection of Neurobiotin into ganglion cells almost always resulted in the labeling of amacrine cells. Taken together, these results suggest a directionality to the movement of tracer across gap junctions connecting amacrine and ganglion cells. We found that the coupling pattern for a given morphological type of cell was generally stereotypic and consistent across retinas. The notable exceptions to this finding were alpha ganglion cells and cells with morphology corresponding to that of on-off direction selective ganglion cells. In both cases, individual cells showed either extensive coupling to both amacrine and ganglion cells or no coupling at all. A notable finding was that, in every case, the neighboring cells within a tracer-coupled array were always within one gap junction of the injected neuron. Furthermore, in many cases, the array formed by the somata of tracer-coupled cells was almost perfectly coincident with the dendritic arbor of the injected cell. Thus, our results indicate that whereas coupling is extensive within the proximal retina, individual cells partake in coupled networks that are stereotypic and highly circumscribed. PMID- 9208997 TI - Synaptic connections between identified neuron types in the antennal lobe glomeruli of the cockroach, Periplaneta americana: II. Local multiglomerular interneurons. AB - Synapses between three types of antennal lobe neurons, namely, local multiglomerular interneurons, antennal receptor neurons, and gamma-aminobutyric acid (GABA)-immunoreactive neurons, were studied by means of a combination of three different markers. The interneurons were labeled by intracellular injection of horseradish peroxidase (HRP) into a single soma or a small group of neurons. Antennal receptor cells were marked by experimentally induced anterograde degeneration, and GABA-containing neurons were identified by postembedding immunogold staining. The following types of connections were found: Local interneurons receive input synapses from 1) degenerated receptor neuron axons, 2) GABA-immunogold-labeled neurites, and 3) non-GABA-immunoreactive neurons. The interneurons form output synapses onto the same three neuron groups. Contacts were also found between HRP-labeled interneurons themselves. The majority of synapses were dyadic. In most cases, only one postsynaptic neuronal process of the dyads was labeled and, thus, was identified. Polysynaptic connections were found between GABA-immunoreactive neurites, HRP-labeled interneuron processes, and nonlabeled neurites or between HRP-labeled interneuron processes and two interconnected GABA-immunoreactive processes. The present findings provide anatomical evidence for an earlier suggested monosynaptic connection between afferent receptor fibers and local, at that time putative, GABAergic interneurons. They further reveal that local multiglomerular interneurons are synaptically interconnected. The interneurons, in addition, form serial connections via more than one GABA-immunoreactive neuron with non-GABA immunoreactive and putative projection neurons. Such polysynaptic connections would be a substrate for a feed-forward "disinhibition" of projection neurons, which has been suggested on the basis of electrophysiological findings. PMID- 9208998 TI - Treatment of plasma refractory thrombotic thrombocytopenic purpura with protein A immunoabsorption. AB - The objective of this study was to assess the effect of protein A immunoabsorption in terms of response rate and toxicities in patients with classical thrombotic thrombocytopenic purpura (TTP) refractory to therapeutic plasma exchange. The study included nine females and one male with a diagnosis of classical TTP treated at multiple university hospital centers with protein A immunoabsorption (PAI) after having failed plasma exchange. The 10 patients had an age range 17-62 years. Prior to PAI, the patients had failed to respond to a mean of 15 (range 6-39) therapeutic plasma exchanges. Three patients had previous episodes of TTP. Evaluation for response to PAI included serial measurements of serum creatinine, lactate dehydrogenase (LDH), hemoglobin, hematocrit, and platelet count before, during, and up to 18 months post-PAI treatment. Seven of 10 study patients had resolution of their TTP. Six of the patients required six or fewer therapeutic PAIs and one required 12 treatments. All responding patients had evidence of improvement by the third PAI treatment. Three patients demonstrated no response to PAI, with two patients expiring from complications of TTP and one patient demonstrating a complete response to a subsequent therapy. No significant toxicity was noted with the use of PAI in this setting. Protein A immunoabsorption in patients with classical TTP refractory to plasma exchange can produce durable complete remissions and warrants comparative studies. PMID- 9209000 TI - Prenatal diagnosis of thalassemia in the Chinese. AB - There is a high prevalence of thalassemia in the Taiwan area. Prenatal diagnosis of severe forms of thalassemia is important for the prevention of this disease. We performed prenatal diagnosis in 167 cases, of which 59 cases were diagnosed by chorionic villi biopsy, 91 cases by amniotic fluid analysis, and 17 cases by cord blood analysis. Hb Bart's hydrops was detected by amplifying the break junction area of the alpha-thalassemia-1 Southeast Asia (SEA)-type gene, and beta thalassemia major was detected by using naturally occurring restriction sites and the amplified created restriction sites (ACRS) method. Screening for hemoglobin (Hb) Bart's hydrops revealed 26 cases of Hb Bart's hydrops, 67 cases of alpha thalassemia-1 (including 6 Hb Bart's hydrops falsely diagnosed as alpha thalassemia-1 from chorionic villi samples), and 38 normal cases. Screening for beta-thalassemia major revealed 8 cases of beta-thalassemia major, 17 cases of beta-thalassemia minor, and 11 normal cases. In cases of alpha-thalassemia, maternal tissue contamination in the chorionic villi samples occurred in the diagnosis of the carrier state and further amniotic fluid analysis will be necessary. There were no any false-positive or false-negative results in beta thalassemia major screening. We conclude that prenatal diagnosis is a reliable and accurate screening method for thalassemia and may be valuable in other areas of high prevalence for thalassemia in Southeast Asia and in Southern China. PMID- 9208999 TI - Resistance to activated protein C in unselected patients with arterial and venous thrombosis. AB - Four hundred and ninety-three consecutive patients referred for arterial or venous thrombosis were screened for congenital and acquired abnormalities of blood coagulation predisposing to thrombosis, and were compared to 341 age- and sex-matched controls. The aim of the study was to determine the prevalence and clinical characteristics of resistance to activated protein C (APC), a defect shown to have different prevalences in different ethnic groups and to be associated with an increased risk of thrombosis. Seventy-three (15%) patients had both APC resistance and the 1691 G to A factor V gene mutation, compared to 6/341 (2%) controls. Seven patients had antithrombin deficiency (1.4%), 11 had protein C deficiency (2.2%), and 4 had protein S deficiency (0.8%). The relative risk of thrombosis in APC-resistant patients was 9.4. Resistance to APC was associated mainly with venous thrombosis, the most frequent being deep-vein thrombosis of the lower limbs. Fifty-eight percent of APC-resistant patients had an associated risk factor at the first thrombotic event: pregnancy and oral contraceptive intake were associated with the first thrombotic episode in 35% and 30% of women, respectively. APC resistance is the most frequent defect of blood coagulation in the general population and in the unselected thrombotic population studied by us. PMID- 9209001 TI - Reinterpretation of G-banded complex karyotypes by fluorescence in situ hybridization with chromosome-specific DNA painting probes and alpha-satellite centromere-specific DNA probes in malignant hematological disorders. AB - In this study, we have performed fluorescence in situ hybridization (FISH) with chromosome-specific DNA painting probes 1, 2, 3, 4, 6, 8, and 12 and centromere specific DNA probes 7,10,12,17,18, and X after G-banding on the same metaphase spreads from four patients with malignant hematological disorders to more precisely interpret their complex karyotypes. The findings demonstrated that the application of combined G-banding and FISH can more accurately explain complex karyotypes of hematological malignancies. FISH can detect not only the origin of marker chromosomes, but also the complex rearrangements that cannot be identified by routine banding techniques. This approach is very important to complement the cytogenetic analysis of malignant disorders and to evaluate the role of chromosome change in the development, progression, and prognosis of tumors. PMID- 9209002 TI - High prevalence of hepatitis C virus infection in patients with B-cell lymphoproliferative disorders in Italy. AB - Starting from the observation that a number of consecutive patients with non Hodgkin's lymphoma (NHL) resulted positive for hepatitis C virus (HCV) antibodies on routine testing, we set up a survey for HCV contact prevalence in all patients with lymphoproliferative disorders (LPD) followed in our institution. We searched for HCV antibodies by a third-generation ELISA technique, followed by a confirmation test (RIBA III); serum viral RNA and HCV genotype were investigated by a RT-PCR technique. We screened a total of 315 patients suffering from B-NHL (91), multiple myeloma (56), MGUS (48), chronic lymphocytic leukemia (57), Waldentrom's macroglobulinemia (13), Hodgkin's disease (HD)(43), and T-NHL (9). While only 1 of 52 patients with a non-B-LPD (HD or T-NHL) had signs of HCV contact (i.e., 1.9%, which is in the range of the normal population in the South of Italy), 59 of 263 patients with a B-LPD (22.4%) had HCV antibodies or RNA, or both, with no major differences among the various types of disorders, except for WM, in which the rate was higher (61.5%). The same prevalence was found for patients tested at diagnosis or during the follow-up, and in transfused or never transfused patients. Only a few patients were aware of having a liver disease; one-half of HCV-positive patients never had transaminase increase. A review of data from Central and Northern Italy is included, showing similar findings; a report from Japan has confirmed such an association, while limited surveys in England have not revealed any correlation. These findings may have important biological and clinical implications. PMID- 9209003 TI - Different hematological phenotypes caused by the interaction of triplicated alpha globin genes and heterozygous beta-thalassemia. AB - The pathophysiology and clinical severity of beta-thalassemia are related to the degree of alpha/non-alpha-chain imbalance. A triplicated alpha-globin gene locus can exacerbate effects of excess alpha-chains caused by a defective beta-globin gene, although this is not observed in all cases. Extensive studies on this condition are lacking. We report a group of 17 patients who are heterozygous for both the alpha alpha alpha(anti-3.7) allele and a mutation in the beta-globin gene cluster. Their clinical phenotypes varied: six had mild anemia with microcytosis and hypochromia, while 11 had more severe anemia with splenomegaly requiring splenectomy (three cases) and blood transfusions (four cases). Different phenotypes were also evident in the presence of the same beta thalassemia mutation: in one family, two individuals had the same alpha- and beta globin genotypes but presented with different hematologic phenotypes. In addition, the complex interaction involving a triplicated alpha-globin gene, beta39- and delta+27-thalassemia mutations is studied in a family with two siblings presenting with hemolytic anemia, normal Hb A2 and increased Hb F. Analysis of this series of patients suggests that additional genetic determinants play a role in modulating phenotypic expression in individuals with identical alpha- and beta-globin genotypes. Interaction with a triplicated alpha-gene can play a role in the clinical presentation of patients with defective beta-globin gene expression and should be considered in the diagnosis of atypical cases. PMID- 9209004 TI - Kinetics of reticulocyte maturity fractions and indices and iron status during therapy with epoetin beta (recombinant human erythropoietin) in cardiac surgery patients. AB - We evaluated the changes in reticulocyte maturity fractions and indices, as measured by flow cytometry, during preoperative treatment with recombinant human erythropoietin (epoetin beta) in cardiac surgery patients. A total of 72 patients was enrolled in this double-blind, randomized, placebo-controlled clinical trial and assigned to the two treatment groups (5 x 500 U/kg bodyweight epoetin beta or placebo intravenously over 14 days preoperatively). Therapy with epoetin beta produced continuous increases in hematocrit/hemoglobin, in the most mature fraction of reticulocytes (LR), and in reticulocyte count. In the first treatment week there were parallel increases in the fraction of most immature reticulocytes (HR) and in the reticulocyte mean cell volume. During the second week of treatment the reticulocyte mean cell hemoglobin content (CHr) decreased, but CHr was independent of all iron parameters, affecting neither the reticulocyte fractions nor the hematocrit/hemoglobin increase. The total preoperative rise in hematocrit correlated with the rises in LR fraction (P = 0.0270) and reticulocyte count (P = 0.0486) during the first week of treatment. Whereas in the epoetin beta patients the preoperative change in HR fraction showed negative correlations with transferrin saturation at baseline (P = 0.0058) and with the preoperative change in iron (P = 0.0113), the preoperative change in the LR fraction correlated positively with transferrin at baseline (P = 0.0115). Postoperatively, the reticulocyte parameters revealed that the onset of increased stimulation of erythropoiesis did not occur in the placebo patients until the second postoperative day, whereas erythropoietic activity in the epoetin beta patients was much higher during the postoperative period as well, as a result of the preoperative stimulation of erythropoiesis. The reticulocyte parameters measured by flow cytometry permitted an objective analysis of erythropoietic activity during treatment with epoetin beta and in all patients postoperatively. Further studies in various types of epoetin beta therapy are needed in order to clarify the value of these reticulocyte parameters for identification of iron deficiency and optimization of epoetin beta treatment regimen. PMID- 9209005 TI - Irreversibly sickled cell beta-actin: defective filament formation. AB - It has been demonstrated that cysteine modification in irreversibly sickled cell beta-actin slows down the remodeling of membrane skeletons [Shartava et al.: J Cell Biol 128:805-812, 1995]. This slow remodeling can be due to alterations in spectrin-actin binding and/or actin-actin interactions in irreversibly sickled cell (ISC) membrane skeletons. In these studies we demonstrate that ISC actin binds spectrin normally. However, ISC beta-actin polymerizes and depolymerizes more slowly than control beta-actin, and forms unusual aggregates when placed under polymerizing conditions. Electron microscopic analysis of actin polymers indicated that ISC actin generates a large amount of aggregates which we conclude are due to the structural modification caused by the disulfide bridge between cysteine284 and cysteine373 in beta-actin. PMID- 9209006 TI - Effect of alpha-thalassemia on sickle-cell anemia linked to the Arab-Indian haplotype in India. AB - Two population groups from Western India with a high prevalence of the beta(S) gene, one tribal (Valsad) and the other nontribal (Nagpur), were studied. The beta(S) gene frequency in both populations was similar (0.22 vs. 0.23), but not the clinical expression of sickle-cell anemia (SS): the sickle homozygotes in the tribal group appeared to have a mild clinical course, whereas the majority in the nontribal group exhibited a more severe clinical phenotype. Both tribal and nontribal SS patients had a similarly high mean hemoglobin (Hb)F expression (18.5% vs. 15.5%) and a high number of F cells (72.3% vs. 66.6%). DNA analysis of the beta-globin gene cluster region revealed that in these two populations, this portion of DNA was identical with and corresponded to the typical Arab-Indian haplotype. Nevertheless, in heterozygotes, the mean beta(S) expression was lower (27.9%) in the tribal as compared to the nontribal group (35.5%). The major epistatic factor distinguishing the milder presentation in tribals vs. a more severe manifestation in nontribals was the very high frequency (0.97) of the alpha-thalassemia gene in the former as compared to the latter (0.24). We conclude that the phenotypic expression of sickle-cell anemia, linked to the Arab India haplotype and expressing similar levels of HbF and F cells, is not uniformly mild in India and that alpha-thalassemia is a powerful and additional epistatic factor in the Indian subcontinent. PMID- 9209007 TI - Lactic acidosis secondary to severe anemia in a patient with paroxysmal nocturnal hemoglobinuria. AB - A patient with paroxysmal nocturnal hemoglobinuria developed lactic acidosis associated with severe anemia. The lactic acidosis corrected after blood transfusion. In the absence of shock, sepsis, or other identifiable causes of lactic acidosis, the severe anemia (hemoglobin 1.2 g/dl) appeared to be the primary etiologic factor. PMID- 9209008 TI - K:Cl cotransport in red cells of transgenic mice expressing high levels of human hemoglobin S. AB - K:Cl cotransport is involved in generating dense red blood cells (RBCs) in homozygotes for HbS (SS). We report on the properties of this transport system in RBCs from control and transgenic mice expressing high levels of human alpha(H) and beta(S) chains. Unlike human SS RBCs, mouse RBCs incubated in isotonic media exhibited a Cl(-)-dependent K+ efflux and therefore have a different set-point for activation. This basal efflux was slightly stimulated by cell swelling to values five times smaller than that in human SS cells; in addition, the delay time for activation was shorter in transgenic than in control mice, but fourfold longer than that of human SS cells. These properties cast doubt on the physiological impact of the mouse K:Cl cotransporter on RBC volume regulation in the mouse and suggest that there are intrinsic differences between the human K:Cl cotransporter and the putative transporter in mice. PMID- 9209009 TI - Low-dose combination chemotherapy for acute myeloid leukemia in elderly patients: a novel approach. AB - Eighteen patients aged 60-84 years with acute myeloid leukemia were treated with low-dose combination chemotherapy comprising cytarabine, etoposide, and mitozantrone or 6-thioguanine; seven of these patients had a pre-existing myelodysplastic syndrome. Nine patients achieved a complete remission, and five had a partial remission. The duration of survival in these 14 responding patients has ranged from 2+ to 19+ months. Myelotoxicity occurred regularly, with time to recovery (neutrophils > or =0.5 x 10(9)/L, platelets > or =50 x 10(9)/L) from nadir being 10-14 days in 12 of the 14 patients. This novel approach with an overall response rate of 78% appears to be a simple and effective form of therapy for elderly patients. PMID- 9209010 TI - Successful treatment of HIV-1-related, zidovudine resistant, thrombocytopenia with didanosine. PMID- 9209011 TI - An ecological study on diet/nutrition and cancer in Japan. AB - Correlation or ecological studies may be useful in identifying cancer risk factors which are distributed relatively homogeneously within a population, but differ greatly between populations or between different periods within a given population. In Japan, the westernization of dietary habits has progressed remarkably in the last few decades and may have resulted in changes of incidence/mortality rates of diet-related diseases, especially cancer. Based on data from the National Nutritional Survey of Japan (1955-1993) and the vital statistics of Japan (1955-1993), we conducted correlation analyses between food/nutrient intake and cancer mortality. The present study on chronological correlations suggests the relationship between westernized dietary habits and mortality by cancers of the colon, breast, ovary and prostate and also the relationship between traditional Japanese dietary habits and mortality by stomach cancer. Our results suggest the importance of diet in the etiology of diet related cancer. Further observation and analyses are needed to confirm the relationship between food/nutrient intake and cancer mortality, while also considering the consumption of tobacco and alcohol and improvement in cancer diagnostic and therapeutic techniques. PMID- 9209012 TI - Diet and stomach cancer in Korea. AB - Stomach cancer is the most prevalent malignant neoplasm in Korea. As of 1991-1992 in Seoul, the cumulative rates reported for the age span 0-74 were 7.6% in males and 3.1% in females. A recent case-control study reported that several food items and cooking methods are associated with increased or decreased risk of stomach cancer among Koreans. An increased risk of stomach cancer was noted among people who frequently consume broiled meats and fishes, salted side dishes (salted/fermented fish products) and salty stewed foods, such as soybean paste thick stew. Frequent consumption of mung bean pancake, tofu, cabbage, spinach and sesame oil decreased the risk. Analysis by cooking method showed that risk of stomach cancer from the same foods varied with preparation. For meat and fish, pan frying was associated with decreased risk, whereas stewing or broiling was associated with increased risk. Pickled vegetables increased the risk, whereas fresh vegetables did not. In a recent cohort study in Seoul, green vegetables and soybean foods were associated with a decreased risk of stomach cancer. Case control and cohort studies have reported that ginseng intake decreased the risk of gastric cancer. PMID- 9209013 TI - Diet and colorectal cancer. AB - Studies of migrants to Australia indicate that the risk of colorectal cancer (CRC) can be influenced by environment during adult life. Under a gene x environment interaction model for the risk of CRC, it is postulated that a high proportion of the variation in CRC incidence among populations is attributable to differences in diet, and that the average genetic susceptibility of populations to dietary carcinogenic components is similar. The possible relevance within populations of individual susceptibility to dietary components is supported by studies showing that the risk of CRC, compared with that of the general population, is not increased in the spouses of cases. It is postulated that, conditional upon a relevant dietary exposure, the level of susceptibility to diet is a major determinant of variation in individual risk. Despite high correlations of meat and fat with CRC among populations, the results of case-control studies have been inconsistent. Inability to stratify subjects by susceptibility may explain the lack of association of intake of fat and meat with CRC in many studies. A new generation of case-control and cohort studies of diet and CRC may extend the limits of epidemiology. Prevention trials with beta-carotene and other compounds have not protected against colorectal neoplasia, and may have exerted adverse effects. This possibility challenges the philosophy of chemoprevention trials in favor of trials of changes in diet towards patterns that are associated with lower risk. PMID- 9209014 TI - Diet, nutrition and prostate cancer. AB - Although much has been written, little is known about the causes of prostate cancer. Variations between populations in the incidence of invasive cancers, together with changes in the incidence of invasive cancers in migrants, have pointed to environmental (lifestyle) factors that might be amenable to intervention. Conversely, there is a lack of international variation in the prevalence of microscopic tumours, so the essential question is: what causes only some of the common microscopic tumours to become aggressive? Dietary factors hold the most promise in this regard and have been the subject of recent reviews. The strongest and most consistent effects are positive associations with animal products such as red meats, eggs and dairy foods, and possibly by implication, fat. Evidence of a protective effect of fruit and vegetables is weak and inconsistent, as is the relationship with vitamin A and carotenoids, such as beta carotene. There are some interesting leads. Lycopene, the carotenoid found in tomatoes, has been reported to be protective; alpha-tocopherol supplementation has shown a protective effect in one intervention study; and vitamin D has been shown to be protective in a prospective study. Interest is also growing in phytoestrogens and the extent to which dietary manipulation with these and other phytochemicals might influence prostate cancer by modifying male sex hormone levels or actions. There is limited evidence of associations with obesity. It is not known whether these are related to a particular dietary pattern or to possible physiological effects on the male's hormonal milieu. Associations with lean body mass are likely to be related to the action of androgens during growth and development. Dietary and nutritional effects on prostate cancer do not appear to be strong, but they may be subtle and attenuated by measurement error. To explore these aspects further will require large prospective studies that include improved (repeated) dietary measurements and also blood sampling, so that genetic polymorphisms can be adequately investigated. Such studies are underway. PMID- 9209015 TI - Chemoprevention: will it work? AB - Cancer chemoprevention has been a major area of inquiry in the United States for over a decade. The field originated from the confluence of developments in laboratory carcinogenesis and nutritional epidemiology. Although still owing much to other research disciplines, chemoprevention researchers have developed models and methodologies specific to the topic. Of particular importance has been the search for intermediate endpoints which might provide valid indications of cancer prevention effectiveness. Intermediate endpoints, however, suffer the limitations of the extrapolation required to the prevention of cancer per se. More direct evidence of chemoprevention effectiveness may come from trials that use actual reduction of cancer incidence as the primary endpoint. Several trials of potential chemopreventive agents are underway or have been completed in the United States or elsewhere with U.S. investigator participation. The results to date have been mixed, and unanticipated findings have raised doubts about the probability of success of primary prevention of cancer by chemoprevention. Future prospects for success may depend upon development of a better understanding of biological mechanisms before progressing to large, long-duration human population trials. PMID- 9209016 TI - Diet and lung cancer 20+ years later: more questions than answers? AB - A critical review of epidemiological studies on diet and lung cancer over the last 20+ years has not provided overwhelming evidence that higher consumption of vegetables, fruit, low-fat/low-cholesterol foods or such micronutrients as carotenoids, selenium and vitamins A, C or E is associated with reduced lung cancer risk. Results from case-control studies have been more positive, with about one half showing fruit and vegetables or their associated micronutrients to be associated with reduced risk. However, most results from cohort and serum micronutrient studies, which avoid the problems of inaccurate accounting of diet and recall bias, were statistically insignificant. Moreover, although most studies were conducted on white male smokers in North America and Europe, the few studies which found significant contrary trends were among subjects of different backgrounds, i.e., black American males and Chinese women in China. Since male smokers vs. nonsmokers in Europe, North America and Japan have been shown in other studies to be lower consumers of fruit/vegetables, and less likely to pursue "perceived healthier lifestyles," the possibility that some of the epidemiological findings on diet and lung cancer are artifactually due to inadequate adjustment for behavioral correlates of smoking and health seekers in a particular society must be considered. This is especially true with recent chemoprevention trials showing higher lung cancer incidence and deaths among consumers of beta-carotene supplements vs. placebo. PMID- 9209017 TI - Chemoprevention research in Europe. AB - The basis for primary prevention of cancer is well-established, because significant causes of cancer are known. However, apart from reducing cigarette smoking, few easily applicable measures to decrease cancer incidence are available. Recently, there has been much interest in chemoprevention in cancer control, with several international bodies, including the International Union Against Cancer (UICC) and the European Union (EU), making important contributions. The results and relevance of these studies are discussed. Many of the problems associated with evaluating cancer prevention strategies generally apply particularly to chemoprevention, and these issues also are addressed. PMID- 9209018 TI - Diet/nutrition and stomach cancer in Japan. AB - Incidence rates of stomach cancer in Japan are much higher than in other countries, but have shown a large decline in the past 20 years. Incidence rates among the Japanese in Hawaii are only one third or less of the indigenous Japanese in Japan. Epidemiologic studies indicate that consumption of salt or salty foods is associated with increased risk of stomach cancer, in concert with the results of experimental studies in rodents. In contrast, consumption of vegetables and fruit is associated with decreased risk. Accumulated evidence is strong that a reduction in salty food intake and an increase in vegetable and fruit intake is important to primary prevention of stomach cancer. Possible protective roles of tea and allium vegetables and supplemental use of micronutrients, interaction between salty food intake and H. pylori infection and other factors need further investigation. Occurrence of stomach cancer in Japan may be reduced by two thirds or more via dietary changes, as seen in the population of Japanese ancestry in Hawaii. PMID- 9209019 TI - Characterization of UreG, identification of a UreD-UreF-UreG complex, and evidence suggesting that a nucleotide-binding site in UreG is required for in vivo metallocenter assembly of Klebsiella aerogenes urease. AB - In vivo urease metallocenter assembly in Klebsiella aerogenes requires the presence of several accessory proteins (UreD, UreF, and UreG) and is further facilitated by UreE. In this study, UreG was isolated and shown to be a monomer with an Mr of 21,814 +/- 20 based on gel filtration chromatography and mass spectrometric results. Although it contains a P-loop motif typically found in nucleotide-binding proteins, UreG did not bind or hydrolyze ATP or GTP, and it exhibited no affinity for ATP- and GTP-linked agarose resins. Site-directed mutagenesis of ureG allowed the substitution of Ala for Lys-20 or Thr-21 in the P loop motif and resulted in the production of inactive urease in cells grown in the presence of nickel; hence, an intact P-loop may be essential for UreG to function in vivo. These mutant cells were unable to synthesize the UreD-UreF-UreG urease apoprotein species that are thought to be the key urease activation complexes in the cell. An insoluble protein species containing UreD, UreF, and UreG (termed the DFG complex) was detected in cells carrying deletions in ureE and the urease structural genes. The DFG complex was solubilized in 0.5% Triton X 100 detergent, shown to bind to an ATP-linked agarose resin, and found to elute from the resin in the presence of Mg-ATP. In cells containing a UreG P-loop variant, the DFG complex was formed but did not bind to the nucleotide-linked resin. These results suggest that the UreG P-loop motif may be essential for nucleotide binding by the DFG complex and support the hypothesis that nucleotide hydrolysis is required for in vivo urease metallocenter assembly. PMID- 9209020 TI - The helicase domain of phage P4 alpha protein overlaps the specific DNA binding domain. AB - Replication initiation depends on origin recognition, helicase, and primase activities. In phage P4, a second DNA region, the cis replication region (crr), is also required for replication initiation. The multifunctional alpha protein of phage P4, which is essential for DNA replication, combines the three aforementioned activities on a single polypeptide chain. Protein domains responsible for the activities were identified by mutagenesis. We show that mutations of residues G506 and K507 are defective in vivo in phage propagation and in unwinding of a forked helicase substrate. This finding indicates that the proposed P loop is essential for helicase activity. Truncations of gene product alpha (gp alpha) demonstrated that 142 residues of the C terminus are sufficient for specifically binding ori and crr DNA. The minimal binding domain retains gp alpha's ability to induce loop formation between ori and crr. In vitro and in vivo analysis of short C-terminal truncations indicate that the C terminus is needed for helicase activity as well as for specific DNA binding. PMID- 9209021 TI - Cloning of the Candida albicans homolog of Saccharomyces cerevisiae GSC1/FKS1 and its involvement in beta-1,3-glucan synthesis. AB - Saccharomyces cerevisiae GSC1 (also called FKS1) and GSC2 (also called FKS2) have been identified as the genes for putative catalytic subunits of beta-1,3-glucan synthase. We have cloned three Candida albicans genes, GSC1, GSL1, and GSL2, that have significant sequence homologies with S. cerevisiae GSC1/FKS1, GSC2/FKS2, and the recently identified FKSA of Aspergillus nidulans at both nucleotide and amino acid levels. Like S. cerevisiae Gsc/Fks proteins, none of the predicted products of C. albicans GSC1, GSL1, or GSL2 displayed obvious signal sequences at their N terminal ends, but each product possessed 10 to 16 potential transmembrane helices with a relatively long cytoplasmic domain in the middle of the protein. Northern blotting demonstrated that C. albicans GSC1 and GSL1 but not GSL2 mRNAs were expressed in the growing yeast-phase cells. Three copies of GSC1 were found in the diploid genome of C. albicans CAI4. Although we could not establish the null mutation of C. albicans GSC1, disruption of two of the three GSC1 alleles decreased both GSC1 mRNA and cell wall beta-glucan levels by about 50%. The purified C. albicans beta-1,3-glucan synthase was a 210-kDa protein as judged by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and all sequences determined with peptides obtained by lysyl endopeptidase digestion of the 210-kDa protein were found in the deduced amino acid sequence of C. albicans Gsc1p. Furthermore, the monoclonal antibody raised against the purified beta-1,3-glucan synthase specifically reacted with the 210-kDa protein and could immunoprecipitate beta-1,3-glucan synthase activity. These results demonstrate that C. albicans GSC1 is the gene for a subunit of beta-1,3-glucan synthase. PMID- 9209023 TI - Isolation by genetic labeling of a new mycobacterial plasmid, pJAZ38, from Mycobacterium fortuitum. AB - In a two-step mating experiment with recipient strains of Mycobacterium smegmatis, the Mycobacterium fortuitum cryptic plasmid pJAZ38 was isolated. Plasmid pJAZ38 was genetically labeled by cointegration formation mediated by the kanamycin-resistant mycobacterial transposon Tn611. The region responsible for replication of pJAZ38 was located and sequenced. This region showed homology with the Mycobacterium avium plasmid pLR7 and the Mycobacterium scrofulaceum plasmid pMSC262, a family of plasmids which have been found to be widespread throughout the mycobacteria. Further experiments showed pJAZ38 to be stably inherited in the absence of selection pressure and compatible with the most commonly used mycobacterial replicon, pAL5000. In contrast to pLR7 and pMSC262, pJAZ38 was able to replicate in M. smegmatis mc(2)155, making it a useful tool for mycobacterial genetics. PMID- 9209022 TI - Characterization of gentamicin 2'-N-acetyltransferase from Providencia stuartii: its use of peptidoglycan metabolites for acetylation of both aminoglycosides and peptidoglycan. AB - The relationship between the acetylation of peptidoglycan and that of aminoglycosides in Providencia stuartii has been investigated both in vivo and in vitro. Adaptation of the assay for peptidoglycan N-->O-acetyltransferase permitted an investigation of the use of peptidoglycan as a source of acetate for the N acetylation of aminoglycosides by gentamicin N-acetyltransferase [EC 2.3.1.59; AAC(2')]. The peptidoglycan from cells of P. stuartii PR50 was prelabelled with 3H by growth in the presence of N-[acetyl-3H]glucosamine. Under these conditions, [3H]acetate was confirmed to be transferred to the C-6 position of peptidoglycan-bound N-acetylmuramyl residues. Isolated cells were subsequently incubated in the presence of various concentrations of gentamicin and tobramycin (0 to 5x MIC). Analysis of various cellular fractions from isolated cells and spent culture medium by the aminoglycoside-binding phosphocellulose paper assay revealed increasing levels of radioactivity associated with the filters used for whole-cell sonicates of cells treated with gentamicin up to 2 x MIC. Beyond this concentration, a decrease in radioactivity was observed, consistent with the onset of cell lysis. Similar results were obtained with tobramycin, but the increasing trend was less obvious. The transfer of radiolabel to either aminoglycoside was not observed with P. stuartii PR100, a strain that is devoid of AAC(2')-Ia. A high-performance anion-exchange chromatography-based method was established to further characterize the AAC(2')-Ia-catalyzed acetylation of aminoglycosides. The high-performance liquid chromatography (HPLC)-based method resolved a tobramycin preparation into two peaks, both of which were collected and confirmed by 1H nuclear magnetic resonance to be the antibiotic. Authentic standards of 2'-N-acetyltobramycin were prepared and were well separated from the parent antibiotic when subjected to the HPLC analysis. By applying this technique, the transfer of radiolabelled acetate from the cell wall polymer peptidoglycan to tobramycin was confirmed. In addition, isolated and purified AAC(2')-Ia was shown to catalyze in vitro the transfer of acetate from acetyl coenzyme A, soluble fragments of peptidoglycan, and N-acetylglucosamine to tobramycin. These data further support the proposal that AAC(2')-Ia from P. stuartii may have a physiological role in its secondary metabolism and that its activity on aminoglycosides is simply fortuitous. PMID- 9209024 TI - The MtrR repressor binds the DNA sequence between the mtrR and mtrC genes of Neisseria gonorrhoeae. AB - Gonococcal resistance to antimicrobial hydrophobic agents (HAs) is due to energy dependent removal of HAs from the bacterial cell by the MtrCDE membrane associated efflux pump. The mtrR (multiple transferrable resistance Regulator) gene encodes a putative transcriptional repressor protein (MtrR) believed to be responsible for regulation of mtrCDE gene expression. Gel mobility shift and DNase I footprint assays that used a maltose-binding protein (MBP)-MtrR fusion protein demonstrated that the MtrR repressor is capable of specifically binding the DNA sequence between the mtrR and mtrC genes. This binding site was localized to a 26-nucleotide stretch that includes the promoter utilized for mtrCDE transcription and, on the complementary strand, a 22-nucleotide stretch that contains the -35 region of the mtrR promoter. A single transition mutation (A- >G) within the MtrR-binding site decreased the affinity of the target DNA for MtrR and enhanced gonococcal resistance to HAs when introduced into HA susceptible strain FA19 by transformation. Since this mutation enhanced expression of the mtrCDE gene complex but decreased expression of the mtrR gene, the data are consistent with the notion that MtrR acts as a transcriptional repressor of the mtrCDE efflux pump protein genes. PMID- 9209025 TI - 6-phospho-alpha-D-glucosidase from Fusobacterium mortiferum: cloning, expression, and assignment to family 4 of the glycosylhydrolases. AB - The Fusobacterium mortiferum malH gene, encoding 6-phospho-alpha-glucosidase (maltose 6-phosphate hydrolase; EC 3.2.1.122), has been isolated, characterized, and expressed in Escherichia coli. The relative molecular weight of the polypeptide encoded by malH (441 residues; Mr of 49,718) was in agreement with the estimated value (approximately 49,000) obtained by sodium dodecyl sulfate polyacrylamide gel electrophoresis for the enzyme purified from F. mortiferum. The N-terminal sequence of the MalH protein obtained by Edman degradation corresponded to the first 32 amino acids deduced from the malH sequence. The enzyme produced by the strain carrying the cloned malH gene cleaved [U 14C]maltose 6-phosphate to glucose 6-phosphate (Glc6P) and glucose. The substrate analogs p-nitrophenyl-alpha-D-glucopyranoside 6-phosphate (pNP alphaGlc6P) and 4 methylumbelliferyl-alpha-D-glucopyranoside 6-phosphate (4MU alphaGlc6P) were hydrolyzed to yield Glc6P and the yellow p-nitrophenolate and fluorescent 4 methylumbelliferyl aglycons, respectively. The 6-phospho-alpha-glucosidase expressed in E. coli (like the enzyme purified from F. mortiferum) required Fe2+, Mn2+, Co2+, or Ni2+ for activity and was inhibited in air. Synthesis of maltose 6 phosphate hydrolase from the cloned malH gene in E. coli was modulated by addition of various sugars to the growth medium. Computer-based analyses of MalH and its homologs revealed that the phospho-alpha-glucosidase from F. mortiferum belongs to the seven-member family 4 of the glycosylhydrolase superfamily. The cloned 2.2-kb Sau3AI DNA fragment from F. mortiferum contained a second partial open reading frame of 83 residues (designated malB) that was located immediately upstream of malH. The high degree of sequence identity of MalB with IIB(Glc)-like proteins of the phosphoenol pyruvate dependent:sugar phosphotransferase system suggests participation of MalB in translocation of maltose and related alpha glucosides in F. mortiferum. PMID- 9209026 TI - Aerobic regulation of the sucABCD genes of Escherichia coli, which encode alpha ketoglutarate dehydrogenase and succinyl coenzyme A synthetase: roles of ArcA, Fnr, and the upstream sdhCDAB promoter. AB - The sucABCD genes of Escherichia coli encode subunits for two enzymes of the tricarboxylic acid (TCA) cycle, alpha-ketoglutarate dehydrogenase (sucAB) and succinyl coenzyme A synthetase (sucCD). To examine how these genes are expressed in response to changes in oxygen and carbon availability, a set of sucA-lacZ, sucC-lacZ, sdhCDAB-sucA-lacZ, and sdhC-lacZ fusions were constructed and analyzed in vivo. While the expression of a sucA-lacZ fusion was low under all cell growth conditions tested, the expression of the sucA gene from the upstream sdhC promoter was considerably higher and varied by up to 14-fold depending on the carbon substrate used. Expression of the sdhCDAB-sucA-lacZ fusion varied by fourfold in response to oxygen. In contrast, no expression was seen from a sucC lacZ reporter fusion, indicating that no promoter immediately precedes the sucCD genes. Taken together, these findings demonstrate that the oxygen and carbon control of sucABCD gene expression occurs by transcriptional regulation of the upstream sdhC promoter. The weaker sucA promoter provides an additional low constitutive level of sucABCD gene expression to supplement transcription from the sdhC promoter. The negative control of sucABCD gene expression seen under anaerobic conditions, like that for the sdhCDAB genes, is provided by the arcA and fnr gene products. These findings establish that the differential expression of eight genes for three of the TCA cycle enzymes in E. coli is controlled from one regulatory element. PMID- 9209028 TI - RpoS- and OxyR-independent induction of HPI catalase at stationary phase in Escherichia coli and identification of rpoS mutations in common laboratory strains. AB - A rapid spectrophotometric assay to determine the activities of HPI and HPII catalases in Escherichia coli extracts has been developed. This assay is based upon the differential heat stabilities of the two enzymes and offers significant advantages over previous methods for quantitation of their activities. Measurement of catalase activities in extracts of various mutant strains confirmed the ability of this method to accurately distinguish the two activities. Contrary to previously published results, HPI catalase activity was observed to increase at stationary phase in strains lacking the stationary-phase sigma factor sigma(s) (RpoS). This increase was independent of OxyR and also occurred in a strain lacking the HPII structural gene, katE. These results suggest a potential novel pathway for HPI induction in response to increased oxidative stress in the absence of HPII. Measurement of HPII activity in strains carrying mutations in pcm (encoding the L-isoaspartyl protein methyltransferase) and surE led to the finding that these strains also have an amber mutation in rpoS; sequencing demonstrated the presence of this mutation in several commonly used laboratory strains of E. coli, including AB1157, W1485, and JC7623. PMID- 9209029 TI - Duplication of the pepF gene and shuffling of DNA fragments on the lactose plasmid of Lactococcus lactis. AB - The gene corresponding to the lactococcal oligopeptidase PepF1 (formerly PepF [V. Monnet, M. Nardi, A. Chopin, M.-C. Chopin, and J.-C. Gripon, J. Biol. Chem. 269:32070-32076, 1994]) is located on the lactose-proteinase plasmid of Lactococcus lactis subsp. cremoris NCDO763. Use of the pepF1 gene as a probe with different strains showed that pepF1 is present on the chromosome of Lactococcus lactis subsp. lactis IL1403, whereas there is a second, homologous gene, pepF2, on the chromosome of strain NCDO763. From hybridization, PCR amplification, and sequencing experiments, we deduced that (i) pepF1 and pepF2 exhibit 80% identity and encode two proteins which are 84% identical and (ii) pepF2 is included in an operon composed of three open reading frames and is transcribed from two promoters. The protein, encoded by the gene located downstream of pepF2, shows significant homology with methyltransferases. Analysis of the sequences flanking pepF1 and pepF2 indicates that only a part of the pepF2 operon is present on the plasmid of strain NCDO763, while the operon is intact on the chromosome of strain IL1403. Traces of several recombination events are visible on the lactose proteinase plasmid. This suggests that the duplication of pepF occurred by recombination from the chromosome of an L. lactis subsp. lactis strain followed by gene transfer. We discuss the possible functions of PepF and the role of its amplification. PMID- 9209027 TI - Aspartate transcarbamylase from the deep-sea hyperthermophilic archaeon Pyrococcus abyssi: genetic organization, structure, and expression in Escherichia coli. AB - The genes coding for aspartate transcarbamylase (ATCase) in the deep-sea hyperthermophilic archaeon Pyrococcus abyssi were cloned by complementation of a pyrB Escherichia coli mutant. The sequence revealed the existence of a pyrBI operon, coding for a catalytic chain and a regulatory chain, as in Enterobacteriaceae. Comparison of primary sequences of the polypeptides encoded by the pyrB and pyrI genes with those of homologous eubacterial and eukaryotic chains showed a high degree of conservation of the residues which in E. coli ATCase are involved in catalysis and allosteric regulation. The regulatory chain shows more-extensive divergence with respect to that of E. coli and other Enterobacteriaceae than the catalytic chain. Several substitutions suggest the existence in P. abyssi ATCase of additional hydrophobic interactions and ionic bonds which are probably involved in protein stabilization at high temperatures. The catalytic chain presents a secondary structure similar to that of the E. coli enzyme. Modeling of the tridimensional structure of this chain provides a folding close to that of the E. coli protein in spite of several significant differences. Conservation of numerous pairs of residues involved in the interfaces between different chains or subunits in E. coli ATCase suggests that the P. abyssi enzyme has a quaternary structure similar to that of the E. coli enzyme. P. abyssi ATCase expressed in transgenic E. coli cells exhibited reduced cooperativity for aspartate binding and sensitivity to allosteric effectors, as well as a decreased thermostability and barostability, suggesting that in P. abyssi cells this enzyme is further stabilized through its association with other cellular components. PMID- 9209030 TI - Divergence in nitrogenases of Azoarcus spp., Proteobacteria of the beta subclass. AB - Nitrogenase is a functionally constant protein catalyzing N2 reduction, which is found in many phylogenetic lineages of Archaea and Bacteria. A phylogenetic analysis of nif genes may provide insights into the evolution of the bacterial genomes. Moreover, it may be used to study diazotrophic communities, when classical isolation techniques may fail to detect all contributing populations. Among six species of the genus Azoarcus, diazotrophic Proteobacteria of the beta subclass, the deduced amino acid sequences of nifH genes of two species were unusually divergent from each other. Nitrogenases of the "authentic" Azoarcus branch formed a monophyletic unit with those of gamma Proteobacteria, thus being in accordance with 16S ribosomal DNA phylogeny. The nitrogenase proteins of the two aberrant strains clustered within the alpha proteobacterial clade with rhizobial nitrogenases. This relationship was supported by bootstrap values of 87 to 98% obtained by various distance and maximum parsimony methods. Phylogenetic distances of NifH proteins indicate a possible lateral gene transfer of nif genes to Azoarcus from a common donor of the alpha subclass at the time of species diversification or several more recent, independent transfers. Application of the phylogenetic analysis to DNA isolated from environmental samples demonstrated novel habitats for Azoarcus: in guts of termites and rice grown in Japan, nifH genes belonging to the authentic Azoarcus branch were detected. This is the first evidence suggesting the occurrence of Azoarcus spp. in a plant other than its originally described host, Kallar grass. Moreover, evidence for expression of nif genes inside grass roots was obtained by in situ hybridization studies with antisense nifH probes. PMID- 9209031 TI - Vacuolar protein sorting in fission yeast: cloning, biosynthesis, transport, and processing of carboxypeptidase Y from Schizosaccharomyces pombe. AB - PCR was used to isolate a carboxypeptidase Y (CPY) homolog gene from the fission yeast Schizosaccharomyces pombe. The cloned S. pombe cpy1+ gene has a single open reading frame, which encodes 950 amino acids with one potential N-glycosylation site. It appears to be synthesized as an inactive pre-pro protein that likely undergoes processing following translocation into appropriate intracellular organelles. The C-terminal mature region is highly conserved in other serine carboxypeptidases. In contrast, the N-terminal pro region containing the vacuolar sorting signal in CPY from Saccharomyces cerevisiae shows fewer identical residues. The pro region contains two unusual repeating sequences; repeating sequence I consists of seven contiguous repeating segments of 13 amino acids each, and repeating sequence II consists of seven contiguous repeating segments of 9 amino acids each. Pulse-chase radiolabeling analysis revealed that Cpy1p was initially synthesized in a 110-kDa pro-precursor form and via the 51-kDa single polypeptide-chain intermediate form which has had its pro segment removed is finally converted to a heterodimer, the mature form, which is detected as a 32 kDa protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. Like S. cerevisiae CPY, S. pombe Cpy1p does not require the N-linked oligosaccharide moiety for vacuolar delivery. To investigate the vacuolar sorting signal of S. pombe Cpy1p, we have constructed cpy1+-SUC2 gene fusions that direct the synthesis of hybrid proteins consisting of N-terminal segments of various lengths of S. pombe Cpy1p fused to the secreted enzyme S. cerevisiae invertase. The N-terminal 478 amino acids of Cpy1 are sufficient to direct delivery of a Cpy1-Inv hybrid protein to the vacuole. These results showed that the pro peptide of Cpy1 contains the putative vacuolar sorting signal. PMID- 9209032 TI - Uridylylation of the P(II) protein in the photosynthetic bacterium Rhodospirillum rubrum. AB - The regulatory protein P(II) has been studied in great detail in enteric bacteria; however, its function in photosynthetic bacteria has not been clearly established. As a number of these bacteria have been shown to regulate nitrogenase activity by a metabolic control system, it is of special interest to establish the role of P(II) in these diazotrophs. In this study, we show that P(II) in Rhodospirillum rubrum is modified in response to the N status in the cell and that addition of ammonium or glutamine leads to demodification. We also provide evidence that P(II) is uridylylated. In addition, we show that not only these compounds but also NAD+ promotes demodification of P(II), which is of particular interest as this pyridine nucleotide has been shown to act as a switch off effector of nitrogenase. Demodification of P(II) by ammonium or NAD+ did not occur in cultures treated with an inhibitor of glutamine synthetase (methionine sulfoximine), whereas treatment with the glutamate synthase inhibitor 6-diazo-5 oxo-norleucine led to total demodification of P(II) without any other addition. The results indicate that P(II) probably is not directly involved in darkness switch-off of nitrogenase but that a role in ammonium switch-off cannot be excluded. PMID- 9209033 TI - Evolutionary relationships among members of the genus Chlamydia based on 16S ribosomal DNA analysis. AB - Nucleotide sequences from strains of the four species currently in the genus Chlamydia, C. pecorum, C. pneumoniae, C. psittaci, and C. trachomatis were investigated. In vitro-amplified RNA genes of the ribosomal small subunit from 30 strains of C. pneumoniae and C. pecorum were subjected to solid-phase DNA sequencing of both strands. The human isolates of C. pneumoniae differed in only one position in the 16S rRNA gene, indicating genetic homogeneity among these strains. Interestingly, horse isolate N16 of C. pneumoniae was found to be closely related to the human isolates of this species, with a 98.9% nucleotide similarity between their 16S rRNA sequences. The type strain and koala isolates of C. pecorum were also found to be very similar to each other, possessing two different 16S rRNA sequences with only one-nucleotide difference. Furthermore, the C. pecorum strains truncated the 16S rRNA molecule by one nucleotide compared to the molecules of the other chlamydial species. This truncation was found to result in loss of a unilaterally bulged nucleotide, an attribute present in all other eubacteria. The phylogenetic structure of the genus Chlamydia was determined by analysis of 16S rRNA sequences. All phylogenetic trees revealed a distinct line of descent of the family Chlamydiaceae built of two main clusters which we denote the C. pneumoniae cluster and the C. psittaci cluster. The clusters were verified by bootstrap analysis of the trees and signature nucleotide analysis. The former cluster contained the human isolates of C. pneumoniae and equine strain N16. The latter cluster consisted of C. psittaci, C. pecorum, and C. trachomatis. The members of the C. pneumoniae cluster showed tight clustering and strain N16 is likely to be a subspecies of C. pneumoniae since these strains also share some antigenic cross-reactivity and clustering of major outer membrane protein gene sequences. C. psittaci and strain N16 branched early out of the respective cluster, and interestingly, their inclusion bodies do not stain with iodine. Furthermore, they also share less reliable features like normal elementary body morphology and plasmid content. Therefore, the branching order presented here is very likely a true reflection of evolution, with strain N16 of the species C. pneumoniae and C. psittaci forming early branches of their respective cluster and with C. trachomatis being the more recently evolved species within the genus Chlamydia. PMID- 9209034 TI - Promoters and transcripts associated with the aroP gene of Escherichia coli. AB - Analysis of in vitro transcriptional events initiating within the region immediately upstream of the aroP coding region has revealed the presence of three promoters, P1, P2, and P3. Both P1 and P2 give rise to mRNA encoding the AroP protein, whereas P3 initiates transcription in the opposite direction. Both P1 and P3 contain UP elements which contribute to promoter strength. Regulation of expression from these three promoters has been examined in vitro by using supercoiled DNA templates and in vivo by using lacZ transcriptional fusions and specific promoter mutants. Expression from P2 is partially repressed by TyrR alone both in vitro and in vivo. Addition of the aromatic amino acid tyrosine, phenylalanine, or tryptophan further increases this repression. P1 is not repressed by TyrR alone but is repressed in vivo in the presence of phenylalanine, tyrosine, or tryptophan. This also occurs in vitro but requires Ca2+ ions in the reaction mixture for its demonstration. Under these conditions, transcription from P3 is enhanced by TyrR protein with phenylalanine, tyrosine, or tryptophan. However, we were unable to demonstrate P3 expression in vivo. Under repressing conditions, there is no production of truncated RNA molecules (from P1), which would be expected if repression involved a roadblock mechanism. PMID- 9209035 TI - Repression of the aroP gene of Escherichia coli involves activation of a divergent promoter. AB - The repression of aroP expression which is mediated by the TyrR protein with phenylalanine, tyrosine, or tryptophan has been shown to be primarily a direct result of TyrR-mediated activation of a divergent promoter, P3, which directs the RNA polymerase away from promoter P1. Evidence which has been presented to support this conclusion is as follows. Repression of P1 does not occur either in vitro or in vivo if wild-type TyrR protein is substituted by the activation negative mutant RQ10 (with an R-to-Q change at position 10). Repression of P1 is greatly diminished if the P3 promoter is inactivated or if a 5-bp insertion is made between the P3 promoter and the binding sites for TyrR. Repression is also abolished if the promoter strength of P1 is increased or a putative UP element associated with P3 is altered. Repression of the second promoter, P2, still occurs if the wild-type TyrR protein is substituted with RQ10 or EQ274. The tryptophan-mediated repression of aroP does not involve the TrpR protein. PMID- 9209036 TI - Phospholipid biosynthesis and solvent tolerance in Pseudomonas putida strains. AB - The role of the cell envelope in the solvent tolerance mechanisms of Pseudomonas putida was investigated. The responses of a solvent-tolerant strain, P. putida Idaho, and a solvent-sensitive strain, P. putida MW1200, were examined in terms of phospholipid content and composition and of phospholipid biosynthetic rate following exposure to a nonmetabolizable solvent, o-xylene. Following o-xylene exposure, P. putida MW1200 exhibited a decrease in total phospholipid content. In contrast, P. putida Idaho demonstrated an increase in phospholipid content 1 to 6 h after exposure. Analysis of phospholipid biosynthesis showed P. putida Idaho to have a higher basal rate of phospholipid synthesis than MW1200. This rate increased significantly following exposure to xylene. Both strains showed little significant turnover of phospholipid in the absence of xylene. In the presence of xylene, both strains showed increased phospholipid turnover. The rate of turnover was significantly greater in P. putida Idaho than in P. putida MW1200. These results suggest that P. putida Idaho has a greater ability than the solvent sensitive strain MW1200 to repair damaged membranes through efficient turnover and increased phospholipid biosynthesis. PMID- 9209037 TI - Identification of two minor subunits in the pilus of Haemophilus influenzae. AB - Haemophilus influenzae type b (Hib) organisms produce pili, which mediate attachment to human cells and are multimeric structures composed of a 24-kDa subunit called pilin or HifA. Although pili from other organisms contain additional proteins accessory to pilin, no structural components other than pilin have been identified in Hib pili. Previous analysis of a Hib pilus gene cluster, however, suggested that two genes, hifD and hifE, may encode additional pilus subunits. To determine if hifD and hifE encode pilus components, the genes were overexpressed in Escherichia coli and the resulting proteins were purified and used to raise polyclonal antisera. Antisera raised against C-terminal HifD and HifE fragments reacted with H. influenzae HifD and HifE proteins, respectively, on Western immunoblots. Western immunoblot analysis of immunoprecipitated Hib pili demonstrated that HifD and HifE copurified with pili. In enzyme-linked immunosorbent assays, antisera raised against a recombinant HifE protein that contained most of the mature protein reacted more to piliated Hib than to nonpiliated Hib or to a mutant containing a hifE gene insertion. Immunoelectron microscopy confirmed that the HifE antiserum bound to pili and demonstrated that the antiserum bound predominantly to the pilus tips. These data indicate that HifD and HifE are pilus subunits. Adherence inhibition studies demonstrated that the HifE antiserum completely blocked pilus-mediated hemagglutination, suggesting that HifE mediates pilus adherence. PMID- 9209038 TI - Bacterial DL-2-haloacid dehalogenase from Pseudomonas sp. strain 113: gene cloning and structural comparison with D- and L-2-haloacid dehalogenases. AB - DL-2-Haloacid dehalogenase from Pseudomonas sp. strain 113 (DL-DEX) catalyzes the hydrolytic dehalogenation of both D- and L-2-haloalkanoic acids to produce the corresponding L- and D-2-hydroxyalkanoic acids, respectively, with inversion of the C2 configuration. DL-DEX is a unique enzyme: it acts on the chiral carbon of the substrate and uses both enantiomers as equivalent substrates. We have isolated and sequenced the gene encoding DL-DEX. The open reading frame consists of 921 bp corresponding to 307 amino acid residues. No sequence similarity between DL-DEX and L-2-haloacid dehalogenases was found. However, DL-DEX had significant sequence similarity with D-2-haloacid dehalogenase from Pseudomonas putida AJ1, which specifically acts on D-2-haloalkanoic acids: 23% of the total amino acid residues of DL-DEX are conserved. We mutated each of the 26 residues with charged and polar side chains, which are conserved between DL-DEX and D-2 haloacid dehalogenase. Thr65, Glu69, and Asp194 were found to be essential for dehalogenation of not only the D- but also the L-enantiomer of 2-haloalkanoic acids. Each of the mutant enzymes, whose activities were lower than that of the wild-type enzyme, acted on both enantiomers of 2-haloacids as equivalent substrates in the same manner as the wild-type enzyme. We also found that each enantiomer of 2-chloropropionate competitively inhibits the enzymatic dehalogenation of the other. These results suggest that DL-DEX has a single and common catalytic site for both enantiomers. PMID- 9209039 TI - Fis is required for illegitimate recombination during formation of lambda bio transducing phage. AB - Specialized transducing particles of phage lambda are formed by illegitimate recombination during prophage induction. We examined the effects of an Esherichia coli int, xis, himA, himD, or fis mutation on illegitimate recombination during formation of lambda Spi- phage, a class of lambda bio transducing phage. This type of phage is distinguishable from the docL and docR particles, which contain one cohesive end and are formed by cutting of the cos site, by plaque formation of lambda bio on Escherichia coli P2 lysogens. The yields of lambda Spi- phage in the int, xis, int-xis deletion, and b2 deletion mutants were about 50- to 200 fold higher than that of the wild-type prophage when bacteria were irradiated with UV light. This result indicates that Int and Xis functions, and the att site, are not required for illegitimate recombination. The yield of lambda Spi- phage in the himA, himD, or fis mutant carrying lambda delta int-xis prophage was 2.6-, 3.3-, or 17-fold lower, respectively, than that in the wild-type bacteria under UV irradiation. Analysis of the nucleotide sequences of the junctions of the transducing phages indicates that recombination at the hotspots, as well as at non-hotspots, takes place between short homologous sequences. Because the growth of infecting phages was not suppressed by the himA, himD, or fis mutation, we conclude that Fis is required, but IHF is only partially required, for short homology-dependent illegitimate recombination during the formation of lambda bio transducing phage. PMID- 9209040 TI - Sequence of xynC and properties of XynC, a major component of the Clostridium thermocellum cellulosome. AB - The nucleotide sequence of the Clostridium thermocellum F1 xynC gene, which encodes the xylanase XynC, consists of 1,857 bp and encodes a protein of 619 amino acids with a molecular weight of 69,517. XynC contains a typical N-terminal signal peptide of 32 amino acid residues, followed by a 165-amino-acid sequence which is homologous to the thermostabilizing domain. Downstream of this domain was a family 10 catalytic domain of glycosyl hydrolase. The C terminus separated from the catalytic domain by a short linker sequence contains a dockerin domain responsible for cellulosome assembly. The N-terminal amino acid sequence of XynC II, the enzyme purified from a recombinant Escherichia coli strain, was in agreement with that deduced from the nucleotide sequence although XynC-II suffered from proteolytic truncation by a host protease(s) at the C-terminal region. Immunological and N-terminal amino acid sequence analyses disclosed that the full-length XynC is one of the major components of the C. thermocellum cellulosome. XynC-II was highly active toward xylan and slightly active toward p nitrophenyl-beta-D-xylopyranoside, p-nitrophenyl-beta-D-cellobioside, p nitrophenyl-beta-D-glucopyranoside, and carboxymethyl cellulose. The Km and Vmax values for xylan were 3.9 mg/ml and 611 micromol/min/mg of protein, respectively. This enzyme was optimally active at 80 degrees C and was stable up to 70 degrees C at neutral pHs and over the pH range of 4 to 11 at 25 degrees C. PMID- 9209041 TI - Temperate Myxococcus xanthus phage Mx8 encodes a DNA adenine methylase, Mox. AB - Temperate bacteriophage Mx8 of Myxococcus xanthus encapsidates terminally repetitious DNA, packaged as circular permutations of its 49-kbp genome. During both lytic and lysogenic development, Mx8 expresses a nonessential DNA methylase, Mox, which modifies adenine residues in occurrences of XhoI and PstI recognition sites, CTCGAG and CTGCAG, respectively, on both phage DNA and the host chromosome. The mox gene is necessary for methylase activity in vivo, because an amber mutation in the mox gene abolishes activity. The mox gene is the only phage gene required for methylase activity in vivo, because ectopic expression of mox as part of the M. xanthus mglBA operon results in partial methylation of the host chromosome. The predicted amino acid sequence of Mox is related most closely to that of the methylase involved in the cell cycle control of Caulobacter crescentus. We speculate that Mox acts to protect Mx8 phage DNA against restriction upon infection of a subset of natural M. xanthus hosts. One natural isolate of M. xanthus, the lysogenic source of related phage Mx81, produces a restriction endonuclease with the cleavage specificity of endonuclease BstBI. PMID- 9209043 TI - Combining localized PCR mutagenesis and natural transformation in direct genetic analysis of a transcriptional regulator gene, pobR. AB - We present a procedure for efficient random mutagenesis of selected genes in a bacterial chromosome. The method combines PCR replication errors with the uptake of PCR-amplified DNA via natural transformation. Cloning of PCR fragments is not required, since mutations are transferred directly to the chromosome via homologous recombination. Random mutations were introduced into the Acinetobacter chromosomal pobR gene encoding the transcriptional activator of pobA, the structural gene for 4-hydroxybenzoate 3-hydroxylase. Mutant strains with strongly reduced PobR activity were selected by demanding the inability to convert 4 hydroxybenzoate to a toxic metabolite. Of spontaneous pobR mutants, 80% carry the insertion element IS1236, rendering them inappropriate for structure-function studies. Transformation with Taq-amplified pobR DNA increased the mutation frequency 240-fold and reduced the proportion of IS1236 inserts to undetectable levels. The relative fidelity of Pfu polymerase compared with Taq polymerase was illustrated by a reduced effect on the mutation frequency; a procedure for rapid assessment of relative polymerase fidelity in PCR follows from this observation. Over 150 independent mutations were localized by transformation with DNA fragments containing nested deletions of wild-type pobR. Sequence analysis of 89 of the mutant pobR alleles showed that the mutations were predominantly single nucleotide substitutions broadly distributed within pobR. Promoter mutations were recovered, as were two mutations that are likely to block pobR translation. One third of the recovered mutations conferred a leaky or temperature-sensitive phenotype, whereas the remaining null mutations completely blocked growth with 4 hydroxybenzoate. Strains containing two different nonsense mutations in pobR were transformed with PCR-amplified DNA to identify permissible codon substitutions. Independently, second-site suppressor mutations were recovered within pcaG, another member of the supraoperonic pca-qui-pob cluster on the Acinetobacter chromosome. This shows that combining PCR mutagenesis with natural transformation is of general utility. PMID- 9209042 TI - Promoter selectivity of Escherichia coli RNA polymerase sigmaF holoenzyme involved in transcription of flagellar and chemotaxis genes. AB - The rpoF gene of Escherichia coli codes for the RNA polymerase sigmaF (or sigma28) subunit, which is involved in transcription of the flagellar and chemotaxis genes. Both sigmaF and sigma70 (the major sigma subunit in growing cells) were overexpressed, purified to homogeneity, and compared with respect to activity and specificity. The affinity of sigmaF to core RNA polymerase (E) is higher than that of sigma70, as measured by gel filtration high-pressure liquid chromatography. In an in vitro transcription system, the holoenzyme (E sigmaF) containing sigmaF selectively transcribed the flagellar and chemotaxis genes, all of which could not be transcribed by E sigma70. This strict promoter recognition property of sigmaF is similar to those of other stress response minor sigma subunits but different from those of the principal sigma subunits, sigma70 and sigma38. sigma70-dependent transcription in vitro is inhibited at high concentrations of all salts tested, showing maximum activity at 50 mM. In contrast, sigmaF-dependent transcription was maximum at 50 mM KCI and then decreased to negligible level at 300 mM; in the cases of potassium acetate and potassium glutamate, maximum transcription was between 200 and 300 mM. DNase I foot printing of the fliC and fliD promoters indicated that sigmaF alone is unable to bind DNA, but E sigmaF specifically recognizes -10 and -35 regions of the sigmaF-dependent promoters with rather long upstream protection. Alteration of the promoter structure after binding of E sigmaF was suggested. PMID- 9209044 TI - Temperature shift experiments with an ftsZ84(Ts) strain reveal rapid dynamics of FtsZ localization and indicate that the Z ring is required throughout septation and cannot reoccupy division sites once constriction has initiated. AB - FtsZ is an essential division protein in bacteria that functions by forming a ring at midcell that mediates septation. To further study the function of the Z ring the effect of a temperature-sensitive mutation, ftsZ84(Ts), on ring dynamics and septal progression was examined. Shifting a strain carrying an ftsZ84(Ts) mutation to the nonpermissive temperature led to loss of Z rings within 1 min. Septal ingrowth was immediately inhibited, and sharply demarcated septa, present at the time of the shift, were gradually replaced by blunted septa. These results indicate that the Z ring is required throughout septation. Shifting filaments to permissive temperature led to a rapid localization of FtsZ84 at regular intervals. Included in these localization events were complete and partial rings as well as spots, although some of these eventually aborted. These results reveal the rapid dynamics of FtsZ localization and indicate that nucleation sites are formed in the absence of FtsZ function. Interestingly, Z rings could not reform at division sites that were constricted although they could reform at sites that had not begun constriction. PMID- 9209045 TI - Characterization of cp18, a naturally truncated member of the cp32 family of Borrelia burgdorferi plasmids. AB - We have mapped the genes encoding the antigenic lipoproteins OspE and OspF to an approximately 18-kb circular plasmid in Borrelia burgdorferi N40. Sequencing and restriction mapping have revealed that this plasmid, cp18, is homologous to an 18 kb region of the cp32 circular plasmids found in the Lyme disease spirochetes. Our data show that cp18 may have arisen from an ancestral cp32 plasmid by deletion of a 14-kb region of DNA, indicating that a significant portion of the cp32 plasmid is not essential in cis for plasmid maintenance. These findings suggest that a relatively small recombinant plasmid capable of being stably maintained in B. burgdorferi could be constructed from a cp32 plasmid. PMID- 9209046 TI - RNA polymerase beta mutations have reduced sigma70 synthesis leading to a hyper temperature-sensitive phenotype of a sigma70 mutant. AB - This work describes a mutational analysis of the interaction between the beta and sigma subunits of Escherichia coli RNA polymerase. The rpoD800 mutant has a temperature-sensitive growth phenotype because the mutant sigma70 polypeptide is not stable at a high temperature. Some rpoB mutations, including rpoB114, enhanced the temperature sensitivity of the rpoD800 mutant. We determined the mechanism by which the rpoB114 rpoD800 double mutant becomes hyper-temperature sensitive for growth. We found that the levels of the mutant sigma70 in the rpoB114 rpoD800 mutant were dramatically reduced compared to that in the rpoD800 mutant after temperature shift-up. The rate of synthesis of the sigma70 polypeptide was reduced in the rpoB114 rpoD800 double mutant compared to the rpoD800 mutant, whereas the half-life of the mutant sigma70 polypeptide after temperature shift-up was the same in both strains. We conclude that because of the reduction of expression of rpoD800 by rpoB114, in concert with the intrinsic instability of the mutant sigma70 polypeptide, the amount of holoenzyme containing sigma70 becomes limiting upon temperature shift-up. This results in the hyper-temperature sensitivity of the rpoB114 rpoD800 double mutant. Furthermore, the effect of rpoB114 on the expression of sigma70 is independent of the rpoD800 allele and is at the transcriptional level. In vitro transcription assays showed that the mutant RNA polymerase RpoB114 was defective in transcribing the two major promoters of the rpoD operon specifically. The effects of these rpoB mutations on gene expression are discussed. PMID- 9209047 TI - Aerobic regulation of isocitrate dehydrogenase gene (icd) expression in Escherichia coli by the arcA and fnr gene products. AB - Isocitrate dehydrogenase, the icd gene product, has been studied extensively regarding the regulation of enzymatic activity and its relationship to the metabolic flux between the tricarboxylic acid cycle and the glyoxylate bypass. In this study, the transcriptional regulation of icd gene expression was monitored by using an icd-lacZ gene fusion and shown to vary over a 15-fold range in response to changes in oxygen and carbon availability. Anaerobic cell growth resulted in fivefold-lower icd-lacZ expression than during aerobic growth. This negative control is mediated by the arcA and fnr gene products. When different carbon compounds were used for cell growth, icd-lacZ expression varied threefold. The results of continuous cell culture studies indicated that this control may be due to variations in cell growth rate rather than to catabolite repression. DNase I footprinting at the icd promoter revealed a 42-bp ArcA-phosphate-protected region that overlaps the start site of icd transcription. Phosphorylation of ArcA considerably enhanced its binding to DNA, while ArcA-phosphate exhibited an apparent dissociation value of approximately 0.1 microM. Based on these studies, ArcA appears to function as a classical repressor of transcription by binding at a site overlapping the icd promoter during anaerobic cell growth conditions. PMID- 9209048 TI - Identification of a monooxygenase from Streptomyces coelicolor A3(2) involved in biosynthesis of actinorhodin: purification and characterization of the recombinant enzyme. AB - The oxidation of phenols to quinones is an important reaction in the oxidative tailoring of many aromatic polyketides from bacterial and fungal systems. Sequence similarity between ActVA-Orf6 protein from the actinorhodin biosynthetic cluster and the previously characterized TcmH protein that is involved in tetracenomycin biosynthesis suggested that ActVA-Orf6 might catalyze this transformation as a step in actinorhodin biosynthesis. To investigate the role of ActVA-Orf6 in this oxidation, we have expressed the actVA-Orf6 gene in Escherichia coli and purified and characterized the recombinant protein. ActVA Orf6 was shown to catalyze the monooxygenation of the tetracenomycin intermediate TcmF1 to TcmD3, strongly suggesting that it catalyzes oxidation of a similar intermediate in actinorhodin biosynthesis. The monooxygenase obeys simple reaction kinetics and has a Km of 4.8 +/- 0.9 microM, close to the figure reported for the homologous enzyme TcmH. The enzyme contains no prosthetic groups and requires only molecular oxygen to catalyze the oxidation. Site-directed mutagenesis was used to investigate the role of histidine residues thought to be important in the reaction; mutants lacking His-52 displayed much-reduced activity, consistent with the proposed mechanistic hypothesis that this histidine acts as a general base during catalysis. PMID- 9209050 TI - Transcriptional and translational regulation of nitrogenase in light-dark- and continuous-light-grown cultures of the unicellular cyanobacterium Cyanothece sp. strain ATCC 51142. AB - Cyanothece sp. strain ATCC 51142 is a unicellular, diazotrophic cyanobacterium which demonstrated extensive metabolic periodicities of photosynthesis, respiration, and nitrogen fixation when grown under N2-fixing conditions. N2 fixation and respiration peaked at 24-h intervals early in the dark or subjective dark period, whereas photosynthesis was approximately 12 h out of phase and peaked toward the end of the light or subjective-light phase. Gene regulation studies demonstrated that nitrogenase is carefully controlled at the transcriptional and posttranslational levels. Indeed, Cyanothece sp. strain ATCC 51142 has developed an expensive mode of regulation, such that nitrogenase was synthesized and degraded each day. These patterns were seen when cells were grown under either light-dark or continuous-light conditions. Nitrogenase mRNA was synthesized from the nifHDK operon during the first 4 h of the dark period under light-dark conditions or during the first 6 h of the subjective-dark period when grown in continuous light. The nitrogenase NifH and NifDK subunits reached a maximum level at 4 to 10 h in the dark or subjective-dark periods and were shown by Western blotting and electron microscopy immunocytochemistry to be thoroughly degraded toward the end of the dark periods. An exception is the NifDK protein (MoFe-protein), which appeared not to be completely degraded under continuous light conditions. We hypothesize that cellular O2 levels were kept low by decreasing photosynthesis and by increasing respiration in the early dark or subjective-dark periods to permit nitrogenase activity. The subsequent increase in O2 levels resulted in nitrogenase damage and eventual degradation. PMID- 9209051 TI - Isolation and characterization of rcsB mutations that affect colanic acid capsule synthesis in Escherichia coli K-12. AB - Regulation of colanic acid polysaccharide capsule synthesis in Escherichia coli requires the proteins RcsC and RcsB, in addition to several other proteins. By sequence similarity, these two proteins appear to be members of the two-component sensor-effector regulatory family found in bacteria. The present study characterizes the functional domains of RcsB. We have isolated mutations in rcsB that are able to suppress an rcsC "up" mutation (i.e., leading to increase in cps transcription) that normally results in constitutive expression of the capsule. In addition, constitutive capsule mutations in rcsB have been isolated. From the characterization of the mutants and by analogy to the three-dimensional structure of CheY, we have begun to define different domains of RcsB and to assign functions to them. A few of the constitutive capsule mutations were localized in an acidic pocket that has been proposed to play a crucial role in phosphorylation of RcsB. As seen in other two-component systems, an aspartate-to-glutamate substitution at the presumed site of phosphorylation of RcsB resulted in constitutive capsule expression. Lastly, several of our rcsB mutants were found to be allele specific (rcsC137 specific) for rcsC, suggesting a physical as well as functional interaction between RcsC and RcsB proteins. PMID- 9209049 TI - epr, which encodes glycylglycine endopeptidase resistance, is homologous to femAB and affects serine content of peptidoglycan cross bridges in Staphylococcus capitis and Staphylococcus aureus. AB - Staphylococcus capitis EPK1 produces a glycylglycine endopeptidase, ALE-1 (M. Sugai, T. Fujiwara, T. Akiyama, M. Ohara, H. Komatsuzawa, S. Inoue, and H. Suginaka, J. Bacteriol. 179:1193-1202, 1997), which hydrolyzes interpeptide pentaglycine chains of cell wall peptidoglycan of S. aureus. Characterizations of the enzyme activity and cloning of ale-1 revealed that ALE-1 is very similar to prolysostaphin produced by S. simulans bv. staphylolyticus. Strain EPK1 is resistant to lysis by ALE-1 and by lysostaphin. A gene that renders the cells resistant to glycylglycine endopeptidase (epr) was found 322 bp upstream of and in the opposite orientation to ale-1. The deduced amino acid sequence of epr showed similarities to FemA and FemB, which have been characterized as factors essential for methicillin resistance of S. aureus. Inactivation of either femA or femB causes decreased resistance to methicillin, increased resistance to lysostaphin, and decreased glycine content in the interpeptide chains of peptidoglycan. Therefore, femAB is suggested to be involved in the addition of glycine to pentapeptide peptidoglycan precursor. S. aureus with epr on a multicopy plasmid had phenotypes similar to those of femAB mutants except that it did not alter resistance level to methicillin. These results suggest that epr and femAB belong to the protein family involved in adding amino acids to the pentapeptide peptidoglycan precursor and that epr is involved in the addition of serine to the pentapeptide. PMID- 9209052 TI - Cloning and analysis of the first cry gene from Bacillus popilliae. AB - An 80-kDa parasporal crystal protein was detected in protein extracts of sporangia of Bacillus popilliae isolated from a diseased larva of the common cockchafer (Melolontha melolontha L.). Amino acid analysis of tryptic peptides revealed significant homology to the Cry2Aa endotoxins of Bacillus thuringiensis. The gene cryBP1 (cry18Aa1), which codes for the parasporal crystal protein, was found in a putative cry operon on the bacterial chromosome, which contains at least one further (smaller) open reading frame, orf1. The 706-amino-acid-long CryBP1 (Cry18Aa1) protein has a predicted molecular mass of 79 kDa and shows about 40% sequence identity to the Cry2 polypeptides of B. thuringiensis. In the light of published observations which suggest that the parasporal crystal proteins of B. popilliae are slightly toxic to their grub hosts, we propose the following survival strategy of B. popilliae. As an obligate pathogen of grubs, B. popilliae germinates in the gut of a grub and the parasporal crystal proteins are released and activated. The activated protein does not cause colloid osmotic lysis but instead damages the gut wall somehow to allow the vegetative cells to enter the hemolymph more easily. By becoming a parasite, B. popilliae can continue to proliferate efficiently while the living grub provides a food supply. This process is in contrast to that of B. thuringiensis, which rapidly kills the insect and is then limited to growth on the larval carcass. PMID- 9209053 TI - Structure/function analysis of the PII signal transduction protein of Escherichia coli: genetic separation of interactions with protein receptors. AB - The PII protein, encoded by glnB, is known to interact with three bifunctional signal transducing enzymes (uridylyltransferase/uridylyl-removing enzyme, adenylyltransferase, and the kinase/phosphatase nitrogen regulator II [NRII or NtrB]) and three small-molecule effectors, glutamate, 2-ketoglutarate, and ATP. We constructed 15 conservative alterations of PII by site-specific mutagenesis of glnB and also isolated three random glnB mutants affecting nitrogen regulation. The abilities of the 18 altered PII proteins to interact with the PII receptors and the small-molecule effectors 2-ketoglutarate and ATP were examined by using purified components. Results with certain mutants suggested that the specificity for the various protein receptors was altered; other mutations affected the interaction with all three receptors and the small-molecule effectors to various extents. The apex of the large solvent-exposed T loop of the PII protein (P. D. Carr, E. Cheah, P. M. Suffolk, S. G. Vasudevan, N. E. Dixon, and D. L. Ollis, Acta Crytallogr. Sect. D 52:93-104, 1996), which includes the site of PII modification, was not required for the binding of small-molecule effectors but was necessary for the interaction with all three receptors. Mutations altering residues of this loop or affecting the nearby B loop of PII, which line a cleft between monomers in the trimeric PII, affected the interactions with protein receptors and the binding of small-molecule ligands. Thus, our results support the predictions made from structural studies that the exposed loops of PII and cleft formed at their interface are the sites of regulatory interactions. PMID- 9209054 TI - Probing interactions of the homotrimeric PII signal transduction protein with its receptors by use of PII heterotrimers formed in vitro from wild-type and mutant subunits. AB - The homotrimeric PII signal transduction protein of Escherichia coli interacts with two small-molecule effectors, 2-ketoglutarate and ATP, regulates two protein receptors, the kinase/phosphatase nitrogen regulator II (NRII) and the glutamine synthetase (GS) adenylyltransferase (ATase), and is subject to reversible uridylylation, catalyzed by the uridylyltransferase/uridylyl-removing enzyme (UTase/UR). The site of PII uridylylation, Y51, is located at the apex of the solvent-exposed T-loop (E. Cheah, P. D. Carr, P. M. Suffolk, S. G. Vasudevan, N. E. Dixon, and D. L. Ollis, Structure 2:981-990, 1994), and an internally truncated PII lacking residues 47 to 53 formed trimers that bound the small molecule effectors but were unable to be uridylylated or activate NRII and ATase (P. Jiang, P. Zucker, M. R. Atkinson, E. S. Kamberov, W. Tirasophon, P. Chandran, B. R. Schefke, and A. J. Ninfa, J. Bacteriol. 179:4342-4353, 1997). We investigated the ability of heterotrimers containing delta47-53 and wild-type subunits to become uridylylated and activate NRII and ATase. Heterotrimers were formed by denaturation and renaturation of protein mixtures; when such mixtures contained a fivefold excess of A47-53 subunits, the wild-type subunits were mostly redistributed into trimers containing one wild-type subunit and two mutant subunits. The resulting population of trimers was uridylylated and deuridylylated by UTase/UR, stimulated the phosphatase activity of NRII, and stimulated adenylylation of GS by ATase. In all except the ATase interaction, the activity of the hybrid trimers was greater than expected based on the number of wild-type subunits present. These results indicate that a single T-loop region within a trimer is sufficient for the productive interaction of PII with its protein receptors. We also formed heterotrimers containing wild-type subunits and subunits containing the G89A alteration (P. Jiang, P. Zucker, M. R. Atkinson, E. S. Kamberov, W. Tirasophon, P. Chandran, B. R. Schefke, and A. J. Ninfa, J. Bacteriol. 179: 4342-4353, 1997). The G89A mutant form of PII does not bind the small-molecule effectors, does not interact with UTase or with NRII, and interacts poorly with ATase. Heterotrimers formed with a 10/1 starting ratio of G89A to wild-type subunits interacted with UTase/UR and ATase to a lesser extent than expected based on the number of wild-type subunits present but activated NRII slightly better than expected based on the number of wild-type subunits present. Thus, intersubunit interactions within the PII trimer can adversely affect the activity of wild-type subunits and may affect the interactions with the different receptors in a variable way. Finally, we formed heterotrimers containing delta47-53 and G89A mutant subunits. These heterotrimers were not uridylylated, did not interact with NRII, and interacted with the ATase only to the extent expected based on the number of G89A subunits present. Thus, the G89A subunits, which contain an intact T-loop region, were not "repaired" by inclusion in heterotrimers along with delta47-53 subunits. PMID- 9209055 TI - The tgl gene: social motility and stimulation in Myxococcus xanthus. AB - Mutations in the tgl locus inactivate social gliding motility in Myxococcus xanthus and block production of pili. The tgl locus is distinctive among the genes for social motility because social gliding and pili can be restored transiently to tgl mutant cells by mixing them with tgl+ cells, a process known as stimulation. The tgl locus was cloned with a linked insertion of transposon Tn5 by using the kanamycin resistance encoded by that transposon. A 16-kb segment of chromosomal DNA complemented the social motility defect when introduced into tgl mutant cells to form a tandem duplication tgl+/tgl heterozygote. To delimit the autonomous tgl transcription unit, subfragments of this 16-kb piece were integrated at the ectopic Mx8 prophage attachment site. A 1.7-kb DNA fragment was identified which, when integrated at the Mx8 site, simultaneously rescued social motility and pilus production. The ability to stimulate tgl mutants was also rescued by the 1.7-kb fragment. Because rescue of stimulation from an mgl deficient donor strain which cannot swarm was observed, this demonstrates that a stimulation donor requires a tgl+ allele but does not require the capacity to swarm actively. The nucleotide sequence of the 1.7-kb fragment revealed two protein coding regions, open reading frame A and open reading frame B (ORFB). ORFB is the tgl gene, because a 613-bp DNA fragment which includes 75% of ORFB rescues tgl-1, -2, and -3 mutants and because disruption of ORFB by deletion or insertion of transposon Tn5lac constitutes a tgl mutation. PMID- 9209056 TI - Identification and localization of the Tgl protein, which is required for Myxococcus xanthus social motility. AB - Tgl protein is required for the production of the type IV pili found at a pole of the Myxococcus xanthus cell. These pili are essential for social motility. Evidence is presented that Tgl is a membrane protein, based on experiments with polyclonal antibody specific for Tgl that was raised against the fusion proteins beta-galactosidase-Tgl and TrpE-Tgl. Immunoaffiity-purified antibody reacted with a protein in M. xanthus having an apparent molecular mass of 27.5 kDa as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, while the sequence of the tgl gene translates into a polypeptide of 27 kDa. Although these numbers are close, it is likely that the primary tgl translation product is processed and modified in M. xanthus. The N terminus has a signal peptidase II recognition sequence, cleavage of which is expected to remove 19 amino acid residues. When the tgl gene is expressed in Escherichia coli, the protein product consistently migrates faster in the gel than mature Tgl expressed in M. xanthus, suggesting a second modification by addition which slows migration of the protein from M. xanthus. Tgl, as detected by its specific antibody, sediments with the membrane fraction of cells. It can be extracted with detergents but not with salt or by the addition of chelators for divalent cations. In an equilibrium gradient, Tgl bands at the buoyant density of membranes and with the NADH-oxidase activity. Intact cells failed to bind anti-Tgl antibody, and less than 2% of the total Tgl is released in soluble form from the periplasm. Yet, cells that had been osmotically shocked and treated with paraformaldehyde were able to react with the specific antibody--a reaction absent from cells with a deletion of the tgl transcription unit. Assuming that osmotic shock disrupts the outer membrane, the fractionation and localization data imply that Tgl is attached to the inner or outer membranes, from which it is exposed to the intermembranous space. Tgl is necessary for synthesis of pili in M. xanthus and is the only pilus protein that can be donated by other cells (stimulation). Tgl contains six tandem copies of the tetratrico peptide repeat structural motif. Its membrane localization, capacity for stimulation, and content of tetratrico structural repeats together suggest that Tgl may be necessary for the assembly of pilin subunits into filaments. PMID- 9209057 TI - Mutational analysis of the linker region of EnvZ, an osmosensor in Escherichia coli. AB - EnvZ, a transmembrane signal transducer, is composed of a periplasmic sensor domain, transmembrane domains, and a cytoplasmic signaling domain. Between the second transmembrane domain and the cytoplasmic signaling domain there is a linker domain consisting of approximately 50 residues. In this study, we investigated the functional role of the EnvZ linker domain with respect to signal transduction. Amino acid sequence alignment of linker regions among various bacterial signal transducer proteins does not show a high sequence identity but suggests a common helix 1-loop-helix 2 structure. Among several mutations introduced in the EnvZ linker region, it was found that hydrophobic-to-charged amino acid substitutions in helix 1 and helix 2 and deletions in helix 1, loop, and helix 2 (delta14, delta8, and delta7) resulted in constitutive OmpC expression. In the linker mutant EnvZ x delta7, both kinase and phosphatase activities were significantly reduced but the ratio of kinase to phosphatase activity increased, consistent with the constitutive OmpC expression. In contrast, the purified cytoplasmic fragment of EnvZ x delta7 possessed both kinase and phosphatase activities at levels similar to those of the cytoplasmic fragment of wild-type EnvZ. In addition, the linker mutations had no direct effect on EnvZ C-terminal dimerization. These results together with previous data suggest that the linker region is not directly involved in EnvZ enzymatic activities and that it may have a crucial role in propagating a conformational change to ensure correct positioning of two EnvZ molecules within a dimer during the transmembrane signaling. PMID- 9209058 TI - An essential function for the phosphate-dependent exoribonucleases RNase PH and polynucleotide phosphorylase. AB - Escherichia coli cells lacking both polynucleotide phosphorylase (PNPase) and RNase PH, the only known P(i)-dependent exoribonucleases, were previously shown to grow slowly at 37 degrees C and to display a dramatically reduced level of tRNA(Tyr)su3+ suppressor activity. Here we show that the RNase PH-negative, PNP negative double-mutant strain actually displays a reversible cold-sensitive phenotype and that tRNA biosynthesis is normal. In contrast, ribosome structure and function are severely affected, particularly at lower temperatures. At 31 degrees C, the amount of 50S subunit is dramatically reduced and 23S rRNA is degraded. Moreover, cells that had been incubated at 42 degrees C immediately cease growing and synthesizing protein upon a shift to 31 degrees C, suggesting that the ribosomes synthesized at the higher temperature are defective and unable to function at the lower temperature. These data indicate that RNase PH and PNPase play an essential role that affects ribosome metabolism and that this function cannot be taken over by any of the hydrolytic exoribonucleases present in the cell. PMID- 9209059 TI - Homoserine O-acetyltransferase, involved in the Leptospira meyeri methionine biosynthetic pathway, is not feedback inhibited. AB - The Leptospira meyeri serovar semaranga metX gene was identified by complementation of an Escherichia coli metA mutant, i.e., devoid of homoserine O succinyltransferase. However, the MetX protein exhibited a homoserine O acetyltransferase activity in agreement with its similarity to homoserine O acetyltransferases. Reverse transcription-PCR analysis demonstrated that metX is the second gene of an operon. PMID- 9209060 TI - Characterization of thiI, a new gene involved in thiazole biosynthesis in Salmonella typhimurium. AB - Thiamine pyrophosphate (TPP) is a required cofactor in Salmonella typhimurium that is generated de novo by the condensation of 4-amino-5-hydroxymethyl pyrimidine (HMP) pyrophosphate and 4-methyl-5-(beta-hydroxyethyl)-thiazole (THZ) monophosphate. The THZ and HMP moieties are independently synthesized, and labeling studies have demonstrated probable metabolic precursors to both. We present herein the initial characterization of thiI, a gene required for THZ synthesis. We show that thiI is a 1,449-bp open reading frame located at minute 9.65 on the S. typhimurium chromosome and that it encodes a 483-amino-acid protein with a predicted molecular mass of 55 kDa. Unlike genes in the thiamine biosynthetic operon at minute 90, thiI is not transcriptionally regulated by TPP. PMID- 9209061 TI - An Escherichia coli host strain useful for efficient overproduction of cloned gene products with NaCl as the inducer. AB - Salt-induced overexpression of genes cloned downstream of the phage T7 phi10 promoter was demonstrated in an Escherichia coli strain (GJ1158) which carries a single chromosomally integrated copy of the gene for phage T7 RNA polymerase under transcriptional control of the cis-regulatory elements of the osmoresponsive proU operon. Plasmids that have been constructed to obtain overproduction of individual target gene products in strain BL21(DE3) (by addition of isopropyl-beta-D-thiogalactopyranoside as an inducer) can directly be transformed into GJ1158. The NaCl induction regimen was also shown to be associated with a decreased propensity for sequestration of overexpressed target proteins within insoluble inclusion bodies. PMID- 9209062 TI - Cloning, sequencing, and regulation of the global nitrogen regulator gene ntcA in the unicellular diazotrophic cyanobacterium Cyanothece sp. strain BH68K. AB - In cyanobacteria, ammonium represses expression of proteins involved in nitrogen fixation and assimilation. The global nitrogen regulator gene ntcA encodes a DNA binding protein, NtcA, that is a transcriptional activator of genes subject to nitrogen control. We report the cloning and sequencing of the ntcA gene from a nitrogen-fixing unicellular cyanobacterium, Cyanothece sp. strain BH68K. The gene comprises 678 nucleotides, and the deduced NtcA protein contains 226 amino acids with a predicted molecular weight of 25,026. In addition, ntcA mRNA levels were measured in cells grown under different nitrogen regimes. Under nitrogen-fixing conditions, ntcA transcripts were weakly expressed. Furthermore, ntcA expression was diminished or inversely proportional to nifHDK expression. Conversely, ntcA expression increased in nitrate-grown cells, and a concentration-dependent increase was seen in ammonium-grown cells up to 1 mM NH4Cl. These results indicate that ntcA is involved more in nitrogen assimilation than in nitrogen fixation and also imply that the rhythmic expression of ntcA and nifHDK transcription may be under the control of a circadian clock. PMID- 9209063 TI - Identification of a fourth gene involved in dTDP-rhamnose synthesis in Streptococcus mutans. AB - We had isolated three genes (rmlA, rmlB, and rmlC) involved in dTDP-rhamnose synthesis in Streptococcus mutans and found that three genes were insufficient for dTDP-rhamnose synthesis (Y. Tsukioka, Y. Yamashita, T. Oho, Y. Nakano, and T. Koga, J. Bacteriol. 179:1126-1134, 1997). The rmlD gene of S. mutans, encoding the enzyme which catalyzes the last step of dTDP-rhamnose synthesis, has been cloned and sequenced. The cell extract of Escherichia coli expressing the rmlD gene of S. mutans exhibited enzymatic activity corresponding to its counterpart in Shigella flexneri, a gram-negative bacterium. Rhamnose was not detected in the cell wall preparation purified from the mutant in which the cloned gene was insertionally inactivated. Rabbit antiserum against S. mutans serotype c-specific antigen did not react with autoclaved extracts from the mutant. The rmlD gene product of S. mutans compensated for the incompleteness of dTDP-rhamnose synthesis by the three previously isolated genes. These results indicate that the rmlD gene product is indispensable for the dTDP-rhamnose pathway and subsequently for the synthesis of serotype-specific antigen in S. mutans. Furthermore, conservation of the rmlD gene in Streptococcus species was demonstrated by Southern blot analysis. PMID- 9209064 TI - Mck1, a member of the glycogen synthase kinase 3 family of protein kinases, is a negative regulator of pyruvate kinase in the yeast Saccharomyces cerevisiae. AB - An interaction between the Saccharomyces cerevisiae protein kinase Mck1 and pyruvate kinase (Pyk1) was detected by using the two-hybrid method. Purified Mck1 was able to phosphorylate purified Pyk1 on Ser in vitro. Pyruvate kinase activity was elevated in mck1 delta cells. Several of the phenotypes of mck1 delta mutants are similar to those observed in cells overexpressing PYK1. Co-overexpression of MCK1 suppressed all of the phenotypes associated with PYK1 overexpression. These results indicate that Mck1 negatively regulates pyruvate kinase activity, possibly by direct phosphorylation. PMID- 9209065 TI - A hot spot in plasmid F for site-specific recombination mediated by Tn21 integron integrase. AB - Integron In2 integrase (IntI1)-mediated site-specific recombination between two primary sites occurs at a high frequency, while that between a primary and a secondary site occurs at frequencies around 10,000 times lower. Secondary sites consist of a pentanucleotide with only two fully conserved residues (GWTMW). The analysis of IntI1-mediated recombinants in the plasmid pOX38 revealed the existence in this plasmid of a site used at a frequency intermediate between those of primary and secondary sites. Analysis of this site showed two potentially relevant structural features: first, a set of two consensus pentanucleotides, separated by 5 bp and in opposite orientations, forming what will be called a double site; and second, a longer sequence with some extent of sequence symmetry with the double site at its 3' end. A recombinant plasmid, pSU18P, containing a double site was constructed. Examination of R388-pSU18P recombinants showed that double sites were used preferentially over single pentanucleotides by IntI1. Comparisons of the nucleotide sequences of known 59-bp elements showed that in most cases there was a double site at each element end. Mutagenesis of the F hot spot was carried out to make it look more like the consensus 59-bp element. The improved sites showed recombination frequencies and specificities almost comparable to those observed at IntI1 primary sites. PMID- 9209066 TI - Versatile insertion plasmids for targeted genome manipulations in bacteria: isolation, deletion, and rescue of the pathogenicity island LEE of the Escherichia coli O157:H7 genome. AB - A system of versatile insertion plasmids was constructed that permits efficient delivery of the target sites of an ultra-rare-cutting endonuclease and the recombinase FLP into preselected sites of the bacterial genome. With the help of this system, the pathogenicity island LEE of the Escherichia coli O157:H7 genome was excised and isolated in vitro, deleted in vivo, rescued as a plasmid, and transferred into another strain. PMID- 9209067 TI - Evolutionary analysis of the hisCGABdFDEHI gene cluster from the archaeon Sulfolobus solfataricus P2. AB - While sequencing the genome of the archaeon Sulfolobus solfataricus P2, we found an 8,313-bp sequence containing a cluster of nine histidine biosynthesis genes in an order different from that of any known his operon. Results of phylogenetic analysis of the coding regions in the putative operon give conflicting evolutionary histories for individual his genes. PMID- 9209068 TI - Purification and sequence analysis of a novel NADP(H)-dependent type III alcohol dehydrogenase from Thermococcus strain AN1. AB - An NADP(H)-dependent alcohol dehydrogenase was isolated from the hyperthermophilic archaeon Thermococcus strain AN1. This enzyme is a homotetramer with a subunit molecular weight of 46,700. The enzyme oxidizes a series of primary linear alcohols but not methanol. The pH and temperature optima with ethanol as the substrate are 6.8 to 7.0 and 85 degrees C, respectively. The enzyme readily reduced acetaldehyde with NADPH as the cofactor. The gene encoding this enzyme has been cloned and sequenced. An open reading frame of 1,218 bp, starting with ATG and ending with TGA, was identified and corresponded to 406 amino acids. Sequence comparisons show that this Thermococcus strain AN1 enzyme has significant homologies with enzymes from the newly defined type III alcohol dehydrogenase family. Thermococcus strain AN1 alcohol dehydrogenase is the first archaeal enzyme belonging to this family. PMID- 9209069 TI - Conservation of the Escherichia coli dnaX programmed ribosomal frameshift signal in Salmonella typhimurium. AB - Escherichia coli DNA polymerase III subunits tau and gamma are produced from one gene, dnaX, by a programmed ribosomal frameshift which generates the C terminal of gamma within the tau reading frame. To help evaluate the role of the dispensable gamma, the distribution of tau and gamma homologs in several other species and the sequence of the Salmonella typhimurium dnaX were determined. All four enterobacteria tested produce tau and gamma homologs. S. typhimurium dnaX is 83% identical to E. coli dnaX, but all four components of the frameshift signal are 100% conserved. PMID- 9209070 TI - A melibiose transporter and an operon containing its gene in Enterobacter cloacae. AB - We detected inducible melibiose transport activity in cells of Enterobacter cloacae IID977. H+, but not Na+, was found to be the coupling cation for this transporter. We cloned and sequenced the gene encoding the melibiose transporter. A homology search of a protein sequence database revealed that this melibiose transporter has high sequence similarity with the lactose transporter (LacY) and the raffinose transporter (RafB) and has some similarity with the melibiose transporter (MelB) of Escherichia coli. PMID- 9209071 TI - Cloning and characterization of the Streptomyces peucetius dnmZUV genes encoding three enzymes required for biosynthesis of the daunorubicin precursor thymidine diphospho-L-daunosamine. AB - Characterization of the dnmZ, dnmU, and dnmV genes from the daunorubicin-producer Streptomyces peucetius by DNA sequence analysis indicated that these genes encode a protein of unknown function plus a putative thymidine diphospho-4-keto-6 deoxyglucose-3(5)-epimerase and thymidine diphospho-4-ketodeoxyhexulose reductase, respectively. Inactivation of each of the three genes by gene disruption and replacement in the wild-type strain demonstrated that all of them are required for daunosamine biosynthesis. PMID- 9209072 TI - Differentiation between cold shock proteins and cold acclimation proteins in a mesophilic gram-positive bacterium, Enterococcus faecalis JH2-2. AB - Transfer of Enterococcus faecalis to a cold temperature (8 degrees C for 4 to 30 h) led to increased expression of 11 cold shock proteins (CSPs). Furthermore, this mesophilic prokaryote synthesized 10 cold acclimation proteins, five of them distinct from CSPs, during continuous growth (4 days) at the same temperature (8 degrees C). PMID- 9209073 TI - Stage-specific requirement of cysteine during in vitro maturation of porcine oocytes for glutathione synthesis associated with male pronuclear formation. AB - The present study was designed to clarify the duration of maturation of porcine oocytes when cysteine promotes male pronuclear (MPN) formation through oocyte glutathione (GSH) synthesis. When cumulus-oocyte complexes (COCs) were cultured in a serum-free maturation medium supplemented or not supplemented with 0.57 mM cysteine, about 90% of oocytes reached metaphase I (M-I) to metaphase II (M-II) and M-II at 36 and 48 h of culture, respectively. When cysteine was added to medium at 0, 12, 24, and 36 h of culture and COCs were cultured for a total of 48 h, oocyte GSH concentrations at the end of culture and the incidence of MPN formation after sperm penetration in vitro were both higher than the values in oocytes cultured for 48 h without cysteine. In contrast, the GSH concentration at 48 h and the incidence of MPN formation were not increased when cysteine was present only during the first 24 h of maturation culture. When cysteine was added to medium every 3 h from 36 h of culture on, a higher incidence of MPN formation was obtained in oocytes cultured in the presence of cysteine from 36 to 42 h than from 36 to 45 h of culture, although GSH concentrations were higher in oocytes cultured with cysteine from 36 to 42 h than from 39 to 42 h of culture. These results suggest that the presence of cysteine in maturation medium is critical only between 42 and 48 h of culture when porcine oocytes are in the late M-I to M II stage of development. At that time cysteine is utilized for GSH synthesis, which is subsequently instrumental in the formation of MPN after sperm penetration in vitro. PMID- 9209074 TI - Expression of tissue inhibitor of metalloproteinase-1 protein and messenger ribonucleic acid by the oviduct of cyclic, early-pregnant, and ovariectomized steroid-treated gilts. AB - It has been suggested that tissue inhibitor of metalloproteinases (TIMP)-1 has a role in reproductive tissues, regulating tissue remodeling or enhancing embryonic development. Oviductal TIMP-1 mRNA levels and protein expression were examined in gilts during the estrous cycle and early pregnancy and in steroid-treated ovariectomized (OVX) gilts by explant culture, two-dimensional SDS-PAGE and fluorography, dot-blot hybridization, immunoblot analysis, RIA, and immunocytochemical studies. TIMP-1 mRNA levels in the oviduct during the estrous cycle were greater (p < 0.02) on Days 2, 15, and 18 than on other days examined, and analysis of oviductal functional segments indicated an effect of day (p < 0.003), an effect of segment (p < 0.007), and a day x segment effect (p < 0.03). The level of TIMP-1 mRNA was greater (p < 0.003) in the isthmus (I) on Day 2 than in the ampulla (A) or infundibulum (INF) or on other days examined (0 and 12). In steroid-treated OVX gilts, an effect of treatment with estradiol valerate (EV) + progesterone (P4) was shown with increased (p < 0.003) TIMP-1 mRNA levels. De novo synthesis of TIMP-1 protein was found throughout the estrous cycle and early pregnancy in all functional segments, but protein expression was greater in the I and greatest on Day 2. In steroid-treated OVX gilts, TIMP-1 protein synthesis was greatest in the I regardless of treatment, but with increased intensity after EV+P4 treatment. TIMP-1 protein was found in oviductal flushings during the estrous cycle and early pregnancy, and in steroid-treated OVX gilts regardless of day, status, or treatment. Differences in TIMP-1 concentrations in oviductal fluid were found by day (p < 0.001), with breed differences detected between the Meishan and standard Western breeds. TIMP-1 protein was immunolocalized primarily to luminal epithelium of the INF, A, and I on all days of the estrous cycle and early pregnancy and to some cells in the stroma and blood vessel walls. Staining intensity correlated with TIMP-1 protein levels in oviductal flushings. The role of TIMP-1 in the oviduct remains to be established. PMID- 9209075 TI - A local oxytocin system is part of the luteinization process in the preovulatory follicle of the marmoset monkey (Callithrix jacchus). AB - Luteinization is a complex differentiation process involving the interaction of extrinsic and intraovarian factors. The aim of this study was to examine the components of an intraovarian oxytocin (OT) system during the periovulatory period in the marmoset monkey, as well as the possible relationship of these components to other factors involved in the luteinization process, using immunohistochemistry and cell culture techniques. Ovaries were collected on Day 7 of the follicular phase (before the endogenous LH surge) and on Day 8 (22 h after an exogenous hCG application, but before ovulation). Before the endogenous LH increase, OT immunoreactivity was detectable at low levels in most antral follicles, where its presence was confined to antral granulosa cell (GC) layers. In contrast, immunoreactivity for the OT receptor (OTR) was localized primarily in the basal GC layer. After application of exogenous hCG, there was a marked enhancement in both the staining intensity and the number of cells positive for OT and the OTR in all GC layers of antral follicles, especially in the preovulatory follicle. Progesterone receptor and 3beta-hydroxysteroid dehydrogenase activity in GC were clearly present only when follicles were obtained after gonadotropin stimulation. Secretion of authentic OT was demonstrated from cultured GC obtained before the LH surge, with highest amounts in cells cultured from preovulatory as opposed to smaller antral follicles. OT production could be stimulated by the application of hCG to the GC cultured from preovulatory follicles, whereas the gonadotropin was without effect on GC from small follicles. FSH had no effect on OT production by GC from either follicle type. Application of OT to the cultures caused an increase in progesterone production by GC from large preovulatory follicles but was without effect on steroidogenesis by cells from small antral follicles. These results describing the presence and distribution of OT and OTR and their modulation by hCG, as well as the luteotrophic effect of OT in cultured GC from preovulatory follicles, implicate OT as a paracrine mediator in the luteinization process in the primate ovary. PMID- 9209076 TI - Time course of pronuclear deoxyribonucleic acid synthesis in parthenogenetically activated bovine oocytes. AB - The progress of pronuclear DNA synthesis was monitored by the radioactive precursor 3H-thymidine during the first cell cycle of parthenogenetically activated bovine oocytes. Bovine oocytes were exposed to Ca2+ ionophore A23187 at 24, 30, or 36 h after the onset of in vitro maturation. Young 24-h oocytes were subsequently cultured for 6 h in the protein synthesis inhibitor, cycloheximide (CHX), to ensure similar rates of activation (96-100%) and pronuclear formation (93-97%) among all groups of oocytes. Subsequent autoradiographic experiments revealed a slightly, but not significantly, accelerated start of DNA synthesis in aged (36 h) oocytes. Maximum levels of DNA labeling were reached within 4 h regardless of oocyte maturation age and persisted for 4 h in 30-h oocytes compared to 2 h in 36-h and 24-h oocytes. The period of DNA synthesis lasted for a total of 12-14 h in all groups of oocytes, and the duration of S-phase was less than 6 h. Since rates of pronuclear formation (58%) and labeling (58%) corresponded to each other, it is argued that only a fully developed pronucleus can synthesize DNA. Oocyte labeling performed in the presence of CHX revealed the capability of CHX to inhibit DNA synthesis up to 8 h postactivation. Removal of CHX by washing when the majority (94%) of oocytes had formed a fully developed pronucleus (at 8 h postactivation) led to the synchronous start of DNA synthesis within 1.5-2 h post-CHX culture. This concomitantly defined the time required for synthesis of vital proteins needed for the entry into S-phase and/or DNA replication. The prolonged exposure of activated oocytes to CHX (10-12 h) negatively affected the pattern of DNA synthesis. The start of DNA synthesis was postponed and reduced pronuclear labeling was observed. In addition, CHX-treated oocytes often exhibited a characteristic punctate pattern of pronuclear labeling in which silver grains were accumulated into clusters. In conclusion, the present results provide knowledge about timing and a possible synchronization of DNA synthesis in parthenogenetically activated bovine oocytes. PMID- 9209077 TI - Comparison of the effects of N-methyl-DL-aspartic acid on gonadotropin and prolactin secretion in anestrous mares and mares exhibiting estrous cycles during anestrus. AB - This study investigated the hypothesis that for a subpopulation of horse mares continuation of estrous cycles during the nonbreeding season may be attributed to continued stimulatory glutamatergic activity on GnRH-secreting neurons. The gonadotropin response to the glutamatergic agonist N-methyl-DL-aspartic acid (NMA) was compared in cycling and anestrous mares during the nonbreeding season. It was anticipated that the gonadotropin response to NMA in cycling mares would be attenuated, compared with that of anestrous mares. The experiment used 16 anestrous mares and 15 mares that cycled during the nonbreeding season. The effect of NMA on prolactin secretion was also evaluated. In addition, the seasonal rhythm of prolactin secretion was compared in anestrous and cycling mares during October-April. In cycling mares, the response to NMA was dependent on the stage of the cycle, and a significantly (p < 0.05) larger proportion responded during the luteal phase (6 of 8), compared with the follicular phase (1 of 7 mares). The proportion of anestrous mares that responded to NMA was similar to that of cycling mares during the luteal phase, but larger than during the follicular phase. In anestrous and cycling mares, NMA suppressed prolactin secretion, and in both groups prolactin secretion decreased during the nonbreeding season. Thus, we conclude that differences in reproductive activity in mares during the nonbreeding season are unlikely to reflect a change in glutamatergic activity. PMID- 9209078 TI - Direction of blood flow and changes in resistance of major arteries supplying the ovary of the pregnant rat. AB - In this study, we examined changes in vascular resistance contributing to increased ovarian blood flow in the pregnant rat. Ovarian blood flow was monitored in vivo using a venous outflow cannulation technique in nonpregnant rats and in pregnant rats at Day 16 and Day 22, and increased from 0.18 +/- 0.02 to 0.81 +/- 0.09 and 1.02 +/- 0.08 ml min(-1) ovary(-1) (mean +/- SEM; n = 7, 7, 6), respectively. Intrinsic vessels within the ovarian complex accounted for 81%, 73%, and 70% of total resistance to ovarian blood flow. Of the two major extrinsic supply vessels, from one-half to two-thirds of the ovarian blood was derived from the uterine artery, and the ovarian artery never contributed to uterine blood flow. These results indicate that the major supply vessels are unlikely to limit ovarian blood flow, even near term when competing demand by the gravid uterus reaches a peak. The finding that ovarian blood flow is derived predominantly from the uterine artery may relate to local mechanisms that influence ovarian function and fetal growth. PMID- 9209079 TI - Synchronization of meiosis in porcine oocytes by exposure to dibutyryl cyclic adenosine monophosphate improves developmental competence following in vitro fertilization. AB - The effect of stage of maturation of the germinal vesicle of porcine oocytes at the time of in vitro maturation on subsequent developmental competence was examined. A large variation exists in the germinal vesicle morphology of oocytes at the time of collection of cumulus-oocyte complexes (COCs) and after culture in the absence of dibutyryl cAMP (dbcAMP) for 20 h. However, the morphology of the germinal vesicle was synchronized to a specific stage after culture in the presence of 1 mM dbcAMP for 20 h. There was no difference in germinal vesicle breakdown rate (total mean, 75.0 +/- 5.4%) or in maturation rate (total mean, 82.1 +/- 2.1 %) at 28 and 44 h of culture, respectively. However, differences in meiotic progress of oocytes were observed (p < 0.05) at 36 h of culture when COCs were exposed to dbcAMP for the first 20 h of maturation, as compared to controls. The incidence of embryos that developed to the blastocyst stage after in vitro fertilization was higher (p < 0.05) when COCs were exposed to dbcAMP (21.5 +/- 2.5%) as compared to controls (9.2 +/- 1.6%). After transfer of experimental embryos to four recipient gilts, the three pregnant recipients delivered 19 live piglets. These results indicate that exposure of COCs to dbcAMP for the first 20 h of culture for maturation increases the homogeneity of oocyte nuclear maturation and improves the efficiency of in vitro production of swine embryos. PMID- 9209080 TI - Hypoxanthine regulation of oocyte maturation in the mouse: insights using hypoxanthine phosphoribosyltransferase-deficient animals. AB - In this study the effects of hypoxanthine (HX) on meiotic maturation were compared using oocytes from mice possessing a hypoxanthine phosphoribosyltransferase null mutation (HPRT-) and from the corresponding HPRT competent background strain (HPRT+). Oocyte-cumulus cell complexes and cumulus cell-enclosed oocytes (oocytes cultured while enclosed by cumulus cells) from HPRT+, but not HPRT-, mice took up HX and contained significant levels of HPRT activity. In addition, FSH increased, and HX suppressed, the de novo synthesis of purines in HPRT+ complexes, whereas de novo synthesis was elevated in HPRT complexes and was unaffected by FSH or HX. After 3 h of HX treatment, lower frequencies of germinal vesicle breakdown (GVB) were observed in cumulus cell enclosed than in denuded HPRT+ oocytes; however, identical frequencies of maturation were observed in denuded and cumulus cell-enclosed HPRT oocytes. This demonstrates a direct inhibitory action of HX on the oocyte that does not depend on salvage, plus an additional action of the cumulus cells that requires HPRT activity. Nevertheless, cumulus cells from HPRT- mice are capable of exerting an additional inhibitory action of dibutyryl cAMP (dbcAMP) on the oocyte. A kinetics analysis of FSH action on HX-arrested cumulus cell-enclosed HPRT+ and HPRT- oocytes revealed, first, that the inhibitory effect of the cumulus cells is transient and, second, that HPRT activity is not required for FSH induction of GVB in HX-arrested oocytes. When dbcAMP- or HX-arrested oocytes were treated with FSH, GVB was blocked to the same extent in HPRT- oocytes with the purine de novo synthesis inhibitor, azaserine, but this drug was less effective in HX-treated HPRT+ oocytes. These results confirm the importance of the de novo pathway in hormone-induced maturation and also support a role for purine salvage as an alternative source of nucleotide in this process. PMID- 9209081 TI - Timing of activation of primordial follicles in mature rats is only slightly affected by fetal stage at meiotic arrest. AB - The objective of this experiment was to test the production-line hypothesis by determining whether timing of activation of primordial follicles at different ages throughout the life of female rats was associated with embryonic age at meiotic arrest of their oogonia. Meiotic arrest of oogonia can occur at any time during mid to late gestation in rats. Pregnant rats received injections of 5 bromo-2'-deoxyuridine (BrdU) on either e17 (treatment 1) or on e19 (treatment 2) (e1 = embryonic day 1 [sperm-positive day 1]). Female offspring (pupping = Day 0) were killed at 30 (6/group), 100 (8/group), 180 (8/group) and 350 (7/group) days of age. Immunohistochemistry was used to determine which primordial follicles contained oocytes and pregranulosa cells that were undergoing DNA synthesis at the time of BrdU injection. The percentage of primordial and primary follicles labeled with BrdU was higher (p < 0.05) at all ages with e17 than with e19 injections. There was a significant (p < 0.001) treatment-by-age interaction in the number of BrdU-containing growing follicles. Growing follicles, however, were found at all ages from both e17 and e19 injections. It is concluded that gestational day of entry into meiotic arrest does influence age of activation of primordial follicles, but it is a minor effect. PMID- 9209082 TI - Pre- and postmeiotic expression of male germ cell-specific genes throughout 2 week cocultures of rat germinal and Sertoli cells. AB - The present study was aimed at examining, by reverse transcription polymerase chain reaction, the expression of germ cell-specific genes in cocultures of Sertoli cells with either pachytene spermatocytes (PS) or round spermatids (RS). In situ hybridization studies showed that the mRNAs encoding phosphoprotein p19 and the testis-specific histone TH2B were specifically expressed in PS whereas those encoding the transition proteins TP1 and TP2 were specific to RS. This resulted in p19:TP1 and TH2B:TP2 ratios that were much higher in PS fractions than in RS fractions prepared by elutriation. When PS or RS were seeded on Sertoli cell monolayers in bicameral chambers, both the number and the viability of the cells decreased during the coculture. However, both parameters were equal to, or higher than, 60% after 2 wk. In PS-Sertoli cell cocultures, the ratios of p19:TP1 and TH2B:TP2 decreased dramatically during the second week of culture; this was due not only to a decrease in the levels of p19 and TH2B mRNAs but also to an enhancement in the relative amounts of TP1 and TP2 as compared to the amounts present on the first day of the coculture. Conversely, both ratios remained low in RS-Sertoli cell cocultures; this was due to a decrease in the levels of the four mRNAs studied during the coculture period. DNA flow cytometry studies showed the occurrence of a haploid cell population (1C) in PS-Sertoli cell cocultures from Day 2 onward, together with a decrease in the tetraploid cell population (4C). No such changes were observed in Sertoli cell-only cultures. By contrast, the haploid population decreased 3-fold during the first week in RS-Sertoli cell cocultures. Immunocytochemical studies demonstrated further that 5-bromo-2'-deoxyuridine-labeled PS of stages V-VIII were able to differentiate into RS under the present coculture conditions. Hence, although clearly imperfect, the present coculture system should help to clarify the local regulations governing spermatogenesis and should allow easier study of spermatogenic cell gene expression. PMID- 9209083 TI - Effects of progesterone on the secondary surge of follicle-stimulating hormone in the rat. AB - In the cyclic rat, the secondary surge of FSH on estrus appears to depend on the LH surge-induced fall in serum concentrations of inhibin. To investigate the involvement of progesterone in the regulation of the secondary surge of FSH, 4 day cyclic rats were treated on proestrus with an antagonist of LHRH (LHRHant) and with an ovulatory dose of ovine (o) LH, progesterone, the antiprogestin RU486, or the combination of RU486 and oLH. Serum concentrations of gonadotropins and inhibin at 1830 h on proestrus and at 0030 h on estrus were determined, and the expression of inhibin/activin subunit mRNAs in the ovary at 0030 h on estrus was analyzed by in situ hybridization. Rats receiving saline showed low expression of alpha-, beta(A)-, and beta(B)-subunit mRNAs in the ovary and low serum levels of inhibin in conjunction with the elevated serum concentrations of FSH on estrus. Administration of LHRHant blocked the decrease in the synthesis and secretion of inhibin and abolished the FSH secondary surge, whereas the injection of oLH prevented these effects. Exogenous progesterone, compared with LHRHant injection, increased alpha-, beta(A)-, and beta(B)-subunit mRNA hybridization intensity in the ovary and serum inhibin immunoreactivity, and also restored, in part, the surge of FSH on estrus. The antiprogestin RU486 did not modify the effect of oLH on either inhibin/ activin subunit mRNAs in the ovary or serum levels of inhibin, but blocked the FSH surge. These results indicate that, in the cyclic rat, 1) the secretion of progesterone on proestrous afternoon, induced by the LH surge, is not involved in the fall of ovarian inhibin synthesis and secretion; and 2) in combination with a drop in serum inhibin, a stimulatory action of progesterone on another factor, possibly pituitary activin, could be necessary to elicit a complete secondary surge of FSH. PMID- 9209085 TI - 1,2-propanediol-induced premature centromere separation in mouse oocytes and aneuploidy in one-cell zygotes. AB - Aneuploidy in germ cells results in reproductive failure and mental and physical disorders in humans. Unfortunately, little is known about the causes and mechanisms of aneuploidy induction. The objective of this study was to test the hypothesis that propylene glycol (1,2-propanediol; PG) induces cytogenetic aberrations in mouse metaphase II (MII) oocytes that predispose zygotes to aneuploidy. Female ICR mice received 7.5 IU eCG and 5.0 IU hCG 48 h later. PG doses of 1300, 2600, and 5200 mg/kg body weight were given 3 h post-hCG; controls received the solvent deionized water. Ovulated oocytes were collected 16 h after administration of PG and processed for cytogenetic analysis. For the one-cell zygote cytogenetic study, females were given PG and paired (1:1) with ICR males for 16 h. Females that mated were given 2 x 10(-3) M colchicine 22 h post-PG, and zygotes were collected 18 h later. PG significantly (p < 0.05) increased both the proportion of MII oocytes with premature centromere separation (PCS) and the proportion of aneuploid one-cell zygotes. These results support the hypothesis that PG-induced PCS in MII oocytes predisposes zygotes to aneuploidy. PMID- 9209084 TI - Activation of porcine oocytes via an exogenously introduced rat muscarinic M1 receptor. AB - Thirty hours after the beginning of in vitro maturation, porcine oocytes were microinjected with mRNA coding for the rat muscarinic M1 receptor. They were then incubated for 15 h to allow sufficient time for completing maturation, translation of the mRNA, and insertion of the receptor into the plasma membrane. They were then treated with acetylcholine, the receptor's agonist, and its effect on inducing various activation-related changes was examined. Acetylcholine treatment triggered the release of Ca2+ from internal stores that could be blocked by atropine, the receptor's antagonist. The Ca2+ release was probably mediated via a G protein, since prior injection of guanosine 5'-O-(2 thiodiphosphate) (GDP-beta-S) totally inhibited the effect of the agonist. Pertussis toxin (PT) had no effect on the Ca2+ transients induced by acetylcholine, suggesting that the signal transduction pathway involved a PT insensitive G protein. Electron microscopy revealed that in the injected oocytes, acetylcholine induced cortical granule exocytosis. The oocytes were released from meiotic arrest as evidenced by the decrease in H1 kinase activity measured in the oocytes during the histone H1 kinase assay. After resuming meiosis they entered interphase: 58.8% of the injected oocytes formed pronuclei after incubation with the agonist. Injection without subsequent acetylcholine treatment, or acetylcholine incubation without prior injection with the receptor mRNA, did not cause these changes. The results provide further evidence that the components of a G protein-mediated signal transduction pathway exist in porcine oocytes and that the activation of this pathway via an exogenously supplied G protein-coupled receptor results in a full complement of oocyte activation events. Whether this pathway transduces the activating signal at sperm-induced oocyte activation requires further examination. PMID- 9209086 TI - Porcine fetal and maternal adrenocorticotropic hormone and corticosteroid concentrations during gestation and their relation to fetal size. AB - A study was conducted to characterize fetal plasma ACTH and corticosteroid concentrations during porcine gestation and to relate plasma corticosteroids to fetal size. Samples were taken in white crossbred pigs at 50, 75, and 100 days of gestation and in Chinese Meishan pigs at Day 75. Fetuses developed in either "crowded" or "roomy" uterine environments after maternal uterine ligation, and all fluid samples were obtained during surgery. Fetal arterial cortisol decreased by 30% between Days 50 and 75 and then increased by 101% between 75 and 100 days. Concomitantly, fetal arterial ACTH increased 4-fold between 50 and 75 days of gestation and 45% between 75 and 100 days. Fetal venous cortisol and ACTH (measured only on Days 75 and 100) concentrations were lower than arterial concentrations. Both amniotic and allantoic fluid cortisol concentrations paralleled those of arterial cortisol but were at least 4-fold less. The percentage of free cortisol on Days 75 and 100 was a constant 24%, whereas cortisol bound to corticosteroid-binding globulin was a constant 60% and albumin bound cortisol was 16%. White crossbred fetal arterial cortisone concentrations were always lower than cortisol concentrations, did not differ between arterial and venous plasma, decreased 50% between 50 and 75 days, and did not change thereafter. Plasma cortisol concentrations in Chinese Meishan fetuses were 30% greater than in white crossbred fetuses of the same age, but plasma ACTH and cortisone did not differ between breeds. Analysis of covariance indicated a negative regression of fetal weight and fetal length on arterial cortisol in white crossbred fetuses only at Day 100, and at Day 75 in Meishan fetuses. Under the specific conditions of this experimental model, these data demonstrate prenatal developmental changes in plasma ACTH and corticosteroids, indicate breed differences in such development, and suggest that a negative relationship exists between endogenous cortisol concentrations and fetal size at specific gestational ages. PMID- 9209087 TI - The estradiol-induced luteinizing hormone surge in the ewe is not associated with increased gonadotropin-releasing hormone messenger ribonucleic acid levels. AB - This experiment was undertaken to determine whether the estrogen-induced LH and GnRH surge in the ewe is associated with activation of a specific subpopulation of neurons in the mid-brain of the ewe as indicated by a change in GnRH mRNA levels. Fifteen ovariectomized ewes were assigned to treatment groups 3-4 wk after ovariectomy. One group of ewes served as controls (n = 2); 50 microg estradiol-17beta (E2) was administered to the remaining ewes. Blood samples were collected from all ewes before treatment (2-h period at 10-min intervals) and continued at 30-min intervals until tissue was collected. At 6, 12, 18, and 24 h after E2 (n = 3 for each time point), brains were collected and processed for localization and measurement of GnRH mRNA by in situ hybridization histochemistry. Serum was analyzed for LH concentrations. Serum LH was pulsatile in controls and decreased at 6 h after E2, and by 12 h the LH surge was initiated. LH levels peaked at 18 h after E2 and returned to basal levels 24 h after E2 treatment. A cRNA probe corresponding to the GnRH-associated peptide region of ovine GnRH prepropeptide mRNA was used to identify GnRH mRNA. Associated with the onset and peak of the LH surge were decreased levels (p < 0.1) of GnRH mRNA in neurons of the preoptic area (POA). Neither the number nor mRNA content of GnRH neurons in the diagonal band of Broca, septal area, or medial basal hypothalamus (MBH) changed during the LH surge. In contrast to E2 induced increases in GnRH secretion during the LH surge, our data indicate that E2 decreases steady-state amounts of GnRH mRNA and that GnRH neurons in the POA are influenced to the greatest extent during the E2-induced GnRH surge. PMID- 9209088 TI - Expression of follistatin and inhibin/activin subunit genes in porcine follicles. AB - Expression of the follistatin (FS) and inhibin/activin (I/A) alpha, beta(A), and beta(B) subunit genes in porcine ovarian follicles was evaluated by reverse transcriptase polymerase chain reaction and/or RNase protection procedures to establish changes during the final stages of follicular development. For the I/A alpha and beta(A) subunits, expression increased (p < 0.05) as follicles progressed to the mid-stage of the follicular phase. The beta(B) subunit was expressed in lower concentrations, and all three I/A subunits showed a marked reduction (p < 0.01) in expression by the late stage of follicular development. In contrast to this pattern, FS gene expression decreased (p < 0.05) as follicles developed from the early (low estradiol) to the mid stage (high estradiol) and continued to decline in advanced follicles (after estrus). The predominant mRNA encoded for FS-315, and the ratio of mRNA for FS-315 to mRNA for FS-288 did not differ significantly during the three stages. Within an animal, concentration of FS mRNAs was related more to stage of the follicular phase than to follicular size. Follicular fluid concentration of FS changed in a manner similar to that observed for expression of its gene. We conclude that expression of the FS gene and translation of its mRNA decrease as follicles approach ovulatory status. PMID- 9209089 TI - Marsupial relaxin: complementary deoxyribonucleic acid sequence and gene expression in the female and male tammar wallaby, Macropus eugenii. AB - The nucleotide and derived amino acid sequences of tammar preprorelaxin were established by combined reverse transcriptase polymerase chain reaction and 3'- and 5'-rapid amplification of cDNA ends methods, using RNA from the corpus luteum of late pregnancy as template. Relaxin gene expression was then investigated in tissues at various stages of the 26-day pregnancy and in adult males. The full length tammar relaxin preprohormone is 579 base pairs. The derived amino acid sequence contains a probable signal peptide of 26 amino acids, a B-domain of 31 amino acids, a C-domain of 111 amino acids, and an A-domain of 24 amino acids, with sequence homologies of 49%, 38%, 47%, and 47%, respectively, to dogfish, pig, and both human relaxins, for the combined A- and B-domains of the functional peptides. The conserved amino acid residues in the B-domain confirm a region shown to be essential for binding of the peptide to its receptor. A relaxin gene is expressed in several other tissues of pregnant tammars including the placenta, follicle, and hypothalamus. Northern analysis showed a 1-kilobase relaxin transcript in the corpus luteum and placenta. Using RNase protection assays, relaxin gene expression in the corpus luteum was greater in early and mid pregnancy, reduced at term, and absent postpartum. These data demonstrate relaxin biosynthesis in both the corpus luteum and placenta in this marsupial and suggest that a relaxin physiology has been conserved during mammalian evolution. PMID- 9209090 TI - Extra- and intracellular effects of divergent selection for pituitary responsiveness to gonadotropin-releasing hormone in prepubertal ram lambs. AB - Divergent selection based on the response of 10-wk-old male lambs to a GnRH challenge has produced two lines of sheep, referred to as high and low lines, that differ in their ability to release LH in response to pharmacological and physiological doses of GnRH. The aim of this study was to determine whether the between-line differences in pituitary sensitivity were related to differences in GnRH receptor number and/or the transduction of the intracellular signal following GnRH receptor activation. Pituitary glands were collected from fourteen 20-wk-old ram lambs from each line, weighed, and sampled for GnRH receptor analysis. The remaining tissue from 9 lambs from each line was dispersed. Of the resultant cell suspension, a sample was stored for measurement of GnRH receptor content and the remainder was plated and cultured for 24 h. The LH responses of cultured cells were measured after exposure to GnRH, A23187, or the phorbol ester phorbol 12,13 dibutyrate (PDB). The results indicated that the pituitary glands of the high line contained significantly higher concentrations of GnRH receptors than did those of the low line and released significantly more LH after stimulation with either GnRH or the Ca2(+)-calmodulin or protein kinase C intracellular second messenger systems. Therefore, the between-line difference in the regulation of pituitary LH secretion occurs at a step distal to the stimulatory sites of action of A23187 and PDB. PMID- 9209091 TI - Reproductive photoresponsiveness in unmanipulated male Fischer 344 laboratory rats. AB - Laboratory rats are considered to be reproductively unresponsive to photoperiod because photoperiod treatments do not induce robust reproductive responses. Groups of 15 young male Fischer 344 (F344) rats were tested for effects of long (16L:8D) and short (8L:16D) photoperiods on testicular development and body mass. Two weeks of short photoperiod inhibited testicular growth, spermatogenesis, and increases in body weight. Testis size became refractory to short photoperiod after 8 wk, but the body weight was lower in short photoperiod for the full 10 wk of the study. In young Harlan Sprague-Dawley rats, in contrast, long and short photoperiod had no effect on either body weight or testis size. Pinealectomized F344 rats had significantly higher body weights and larger testes than did sham operated controls, suggesting that the effects of photoperiod are mediated, at least in part, by the pineal gland. The F344 strain of laboratory rats is the first in which unmanipulated animals have been found to be robustly affected by photoperiod, indicating that this strain could be a valuable new model for the study of reproductive regulation by photoperiod. PMID- 9209092 TI - Pituitary gonadotrophs are strongly activated at the beginning of spermatogenesis in African catfish, Clarias gariepinus. AB - Pituitary gonadotrophs were studied in male African catfish between 1 and 37 wk of age using antisera against the LH subunits for immunohistological and radioimmunological purposes, and cRNA probes for in situ hybridization. Immunoreactive material was already detectable at the earliest age examined. In juveniles, the signal for the common glycoprotein alpha subunit (GP alpha) was stronger than that for the LH beta subunit. Accordingly, an excess of radioimmunoassayable GP alpha 100 times that of LHbeta was recorded in the pituitary. Using in situ hybridization, the mRNAs were detected 7 (GP alpha) and 13 (LHbeta) wk after hatching. Detection of LHbeta mRNA coincided with a 300-fold increase in the pituitary content of LHbeta and intact LH, whereas GP alpha increased only 15-fold. The number of gonadotrophs per pituitary and the amount of LH per gonadotroph also increased strongly. The strong, initial increase in pituitary LH levels was always associated with the presence of spermatocytes. However, in a limited number of cases (3 out of 12 fish), the pituitary LH content was low despite the presence of spermatocytes. The number of gonadotrophs, the staining intensities (reflecting protein and mRNA), and the pituitary LH content kept increasing, although at a reduced rate, until completion of the first wave of spermatogenesis. In view of the excess of GP alpha over LHbeta, we conclude that expression of the two subunits is regulated in part by different mechanisms, and that expression of LHbeta is rate-limiting for the amount of intact LH. The strong activation of the gonadotrophs shortly after meiosis opens the possibility that a signal of testicular origin stimulates LH expression, in particular its beta subunit. In the absence of a FSH-like gonadotropin in catfish, we propose that LH covers all functions requiring gonadotropic regulation in the African catfish. PMID- 9209093 TI - Changes in the forward and reverse metabolism of aromatizable androgens during the development of large ovarian cysts in the pregnant rat. AB - Twice-daily treatments with subovulatory doses of hCG result in the development of large ovarian cysts that possess the capacity to produce preovulatory amounts of estradiol (E2) in the presence of exogenous substrate (Biol Reprod 1991; 45:34 42). To determine the effects of prolonged stimulation by subovulatory doses of LH-like activity on the ability of ovarian follicles to produce aromatizable androgens and their metabolites and on the capacity of similar follicles to metabolize exogenous androstenedione (A4) and testosterone, pregnant rats were treated with either 0 (control), 1, or 3 IU hCG twice daily for 9 days, beginning on Day 13 of pregnancy. The largest follicles or cysts in the ovaries of these animals on Days 15, 17, 19, and 22 were incubated for 4 h in the presence of 1) medium alone, 2) 1 mM cAMP, 3) 800 ng/ml A4 with or without cAMP, or 4) 800 ng/ml testosterone with or without cAMP. In the presence or absence of cAMP, follicular incubates from controls displayed limited amounts of A4, testosterone, E2, estrone (E1), and 5alpha-reduced androgen accumulation compared to incubates from rats treated with hCG. By Day 22, follicular incubates from rats treated with 3 IU hCG contained more A4 than incubates from animals treated with 1 IU hCG. However, on each day tested, incubates from rats treated with 1 IU hCG displayed at least as much, and usually more, testosterone, E2, E1, and 5alpha-reduced androgens as did incubates from rats treated with 3 IU hCG. Follicles from rats treated with 3 IU hCG lost their ability to respond to cAMP with increased steroidogenesis. The capacity of follicles from controls and from rats treated with 3 IU hCG to metabolize exogenous A4 to testosterone, E2, and 5alpha-reduced androgens was maintained between 32 and 37 ng of steroid/4 h from Day 15 to Day 22, whereas the capacity of follicles from rats treated with 1 IU hCG to metabolize A4 ranged from 68 to 92 ng of steroid/4 h. Similar patterns for E2 and 5alpha-reduced steroid production were observed when exogenous testosterone was used as substrate. Follicles from all in vivo treatment groups displayed similar capacities to reverse-metabolize exogenous testosterone to A4 and E1 on Day 15 of pregnancy. This capacity increased dramatically, from 29 to 75 ng of steroid/4 h, between Days 15 and 17 for follicles from rats treated with 3 IU hCG. A similar increase was not observed for follicular incubates from rats treated with 1 IU hCG until Day 22 of pregnancy, and was never observed for follicles from controls. In summary, prolonged exposure to stimulation by subovulatory doses of LH-like activity differentially affects the forward and reverse metabolism of aromatizable androgens by ovarian cysts that are developing in the pregnant rat. The limited ability of large, hCG-induced cysts to forward-metabolize A4 and the apparent increased capacity of these follicles to reverse-metabolize testosterone to A4 indirectly support the notion that follicular 17-ketosteroid reductase and 17beta-hydroxysteroid dehydrogenase activities may be differentially regulated during the induction of ovarian cysts in response to prolonged stimulation by gonadotropins in the anovulatory environment of the pregnant rat. PMID- 9209094 TI - Homologous down-regulation of luteinizing hormone/chorionic gonadotropin receptors by increasing the degradation of receptor transcripts in human uterine endometrial stromal cells. AB - We investigated the possible homologous down-regulation of LH/hCG receptors in human uterine endometrial stromal cells. The cells contained a major 4.3-kilobase (kb) and minor 3.6-kb, 2.4-kb, 1.8-kb, and 1.0-kb transcripts of receptors and an 80-kDa receptor protein that can bind [125I]hCG. Culturing these cells with increasing concentrations of highly purified hCG resulted in a dose-dependent significant decrease in steady-state levels of all the receptor transcripts, the 80-kDa receptor protein, and [125I]hCG binding as compared to the control values. The hCG effect was hormone specific and required the conformation of native hormone. The decrease in steady-state receptor transcript levels by hCG was not due to a decrease in the transcription rate of the LH/hCG receptor gene. It was rather due to a significant decrease in the half-life of receptor transcripts from 40.1 +/- 12.4 h in the control to 13.3 +/- 3.6 h after treatment. The homologous down-regulation observed in the present study may potentially explain low endometrial receptor levels in the postmenopausal human endometrium. PMID- 9209096 TI - Age-related changes in the photoperiodic response of Siberian hamsters. AB - Previous studies indicate that as Siberian hamsters (Phodopus sungorus) age, they may lose their ability to show gonadal regression in response to short days. In one study, hamsters that regressed on short days early in life failed to regress when exposed to short days a second time later in life. Thus, Siberian hamsters may experience age-related deficits in photoresponsiveness or may be incapable of regressing twice. In the present study, we attempted to discriminate between these possibilities by examining patterns of gonadal regression in hamsters transferred back and forth from long (16L:8D) to short days (6L:18D) every 6.5, 13, or 26 wk for a 2-yr period. A control group was maintained on long days and had enlarged gonads throughout the entire study. Hamsters alternating between 26 wk of long and short days exhibited complete gonadal regression during their initial but not during their second exposure to short days. Hamsters alternated between long and short days every 13 or 6.5 wk showed regression two to three times, respectively. After about 52 wk of age, the majority of animals in both groups did not regress when exposed to short days. Taken together, the results of this experiment indicate that male Siberian hamsters 1) can exhibit at least two rounds of short-day-induced gonadal regression and 2) fail to regress on short days after about 1 yr of age. PMID- 9209095 TI - Effects of equine chorionic gonadotropin, human chorionic gonadotropin, and laparoscopic artificial insemination on embryo, endocrine, and luteal characteristics in the domestic cat. AB - The effects of gonadotropin treatment and laparoscopic artificial insemination (AI) on embryo quality, serum progesterone and estradiol concentrations, and luteal progesterone content were examined in the domestic cat. These data were compared to similar historical data reported for naturally estrual, mated queens. All queens in this study (n = 32) were treated with eCG followed by 1) natural breeding (eCG-NB), 2) NB and hCG (eCG-NB-hCG), 3) NB and a sham AI procedure (eCG NB-sham AI), or 4) hCG and actual AI (eCG-hCG-AI). Queens ovulating in response to treatment were ovariohysterectomized, and oviducts and uteri were flushed to collect embryos. Ovarian structures were recorded, corpora lutea (CL) were excised and evaluated for progesterone content, and serum was analyzed for estradiol-17beta and progesterone. Follicle and CL numbers ranged from 0 to 28 and 2 to 42 per cat, respectively, and treatment means did not differ (p > or = 0.05) among groups. Embryos were recovered from oviducts and uterine horns in all treatment groups, and recovery ranged from 60-96%. Mean embryo number per queen ranged from 8.2 +/- 2.6 to 23.2 +/- 3.8 and did not differ (p > or = 0.05) among groups. However, the proportions of unfertilized oocytes were greater (p < 0.05) for groups treated with hCG and/or artificially inseminated, and the proportion of blastocysts produced (31 of 107, 29.0%) was lower (p < 0.05) in the eCG-hCG-AI group than for any other treatment (range, 59 of 116 [50.9%] to 67 of 116 [57.8%]). Not all queens in each group produced good-quality embryos (eCG-NB, 5 of 5; eCG-NB-hCG, 5 of 8; eCG-NB-sham AI, 2 of 5; and eCG-hCG-AI, 3 of 6). Serum progesterone and estradiol-17beta, and total luteal progesterone per ovary did not differ (p > or = 0.05) among treatments. Compared to historical controls (naturally estrual, mated queens), eCG-NB queens produced > 4 times as many good quality embryos and blastocysts. Similarly, eCG-hCG-AI-treated queens produced > 4 times the number of oocytes and embryos, although a high proportion of these were poor quality and did not develop to blastocysts. Together, these results indicate that queens treated with eCG are capable of consistently producing many good-quality embryos, at least half of which develop to blastocysts in culture. These data support the use of eCG in felids and suggest that other factors are responsible for reduced pregnancy success and small litter sizes following assisted reproduction. PMID- 9209097 TI - Immunoblot detection of decreased antibodies to haptoglobin-like protein in the serum of infertile women with or without endometriosis. AB - The purpose of this study was to find out differences in the levels of antibodies to distinct antigens in the serum of fertile versus infertile patients with and without endometriosis and to identify these antigens. Blood was collected from 61 patients undergoing laparoscopy for pelvic pain, infertility, or tubal ligation. Serum antibodies against serum antigens with apparent molecular masses of 22 kDa and 18 kDa were assessed by immunoblot analysis. Gel filtration, HPLC DEAE ion exchange chromatography, NH2-terminal sequencing, and double immunodiffusion were used to characterize and identify these proteins. The relative amount of antibodies reacting with 22- and 18-kDa proteins detected in a standard preparation of antigens was significantly lower in the serum of infertile patients with endometriosis (0.20 +/- 0.05 and 0.57 +/- 0.10) and without endometriosis (0.21 +/- 0.06 and 0.53 +/- 0.08) compared to that of control fertile women without endometriosis (0.53 +/- 0.08 and 1.09 +/- 0.13). After purification by chromatography, the NH2-terminal amino acid sequence of the proteins in the 22- and 18-kDa range was identical through 20 amino acids with the alpha chain of the human haptoglobin. Double immunodiffusion implied immunochemical identity between commercial human haptoglobin and the purified proteins. We conclude that infertile patients with and without endometriosis show reduced serum levels of antibodies against a haptoglobin-like protein. These results would indicate an alteration of the immune system or changes in the levels of these antigens in infertility and/or endometriosis. PMID- 9209098 TI - Identification, expression, and regulation of the transcriptional factor polyomavirus enhancer activator 3, and its putative role in regulating the expression of gamma-glutamyl transpeptidase mRNA-IV in the rat epididymis. AB - Gamma-glutamyl transpeptidase (GGT) mRNA-IV is highly expressed in the initial segment of the rat epididymis and is regulated by testicular factors. The promoter region for GGT mRNA-IV contains five conserved polyomavirus enhancer activator 3 (PEA3)-binding motifs (5'-AGGAAG-3'). We hypothesize that PEA3 is present in the rat epididymis and is regulated by one or more testicular factors. Western blot analyses showed that a 62-kDa protein was detected in the nuclear extract from the rat initial segment at higher levels than in the distal epididymal regions. Electrophoretic mobility shift assays (EMSAs) showed that the nuclear extract specifically bound to the PEA3 motif, forming a DNA-protein complex. This complex contained the 62-kDa PEA3 protein as demonstrated by EMSAs and Southwestern analyses. Northern blot analyses and RNase protection analyses showed that PEA3 mRNA was predominantly expressed in the initial segment as compared to the distal epididymal regions and was under the regulation of testicular factors. These results suggest that PEA3 could be involved in the regulation of expression of the rat GGT mRNA-IV gene in response to testicular factors in the initial segment. PMID- 9209099 TI - Expression and function of the c-kit proto-oncogene protein in mouse sperm. AB - The presence and role of the c-kit protein were examined in mature sperm of the mouse. Monoclonal antibodies (mAbs) against the c-kit protein were used to perform immunohistochemical staining, electron microscopy studies, and Western blot analysis. The acrosomal region of both fixed and unfixed noncapacitated sperm stained with the antibodies. No acrosomal staining was noted in acrosome reacted (AR) sperm. Electron microscopy studies demonstrated immunogold label on the plasma membrane of the acrosome and confirmed the lack of binding following the acrosome reaction. Proteins corresponding to 33 kDa, 48 kDa, and 150 kDa were detected by the antibodies utilizing Western blot analysis. The 48-kDa and 150 kDa proteins were released into the media during sperm capacitation, and release from the acrosome was dependent upon the acrosome reaction. The mAbs significantly inhibited the acrosome reaction and increased sperm agglutination. Monoclonal antibody ACK1 significantly inhibited the motility of the sperm, whereas mAbs ACK2 and NCL-ckit did not. These results suggest that c-kit-related proteins are present in mature sperm and may play a role in capacitation and/or the acrosome reaction. PMID- 9209100 TI - Transcription and translation in bovine nuclear transfer embryos. AB - The development of bovine embryos reconstructed by nuclear transfer (NT) is poor compared to that of embryos produced by in vitro fertilization. One reason for this could be incomplete reprogramming of the transferred nucleus. Therefore, with a view to optimizing the conditions for NT, the reprogramming of blastomere nuclei from 16- to 32-cell-stage in vitro-fertilized (IVF) embryos was investigated following NT by fusion of individual blastomeres with cytoplasts prepared from oocytes at two different stages of maturation. Heterogeneous RNA (hnRNA) production, nucleolar ultrastructure, and protein profiles of the NT embryos up to the 8-cell stage were analyzed. In all NT embryos analyzed for their hnRNA production (n = 133), [3H]uridine incorporation was higher at the 1-, 2-, and 4-cell stages than in control IVF embryos (n = 50). Ultrastructural examination of 11 NT embryos revealed evidence of transcriptional activity; fibrillar and granular components were seen in the nucleolus at the 1-cell stage. At the 2-, 4-, and 8-cell stages, fibrillar components were still evident but granular components had become scarce. The hnRNA synthesis, however, was not reflected in the one-dimensional electrophoretic patterns of protein production in the NT embryos (n = 56); these were largely similar to those of IVF embryos (n = 34) of corresponding stages. Thus, NT embryos made in this way do not behave like equivalent IVF embryos, suggesting that reprogramming of the transferred nucleus is absent or incomplete. PMID- 9209101 TI - Inhibition of nitric oxide synthase enhances peripheral nerve regeneration in mice. AB - We tested the hypothesis that inhibition of nitric oxide synthase (NOS) following transection of the sciatic nerve in the mouse would adversely influence regeneration of myelinated fibers from the proximal stump. NOS was inhibited by N(omega)-nitro-L-arginine-methyl ester (L-NAME; 10 mg/kg i.p.), a broad spectrum NOS inhibitor given twice daily for the first 10 days following nerve transection in Swiss mice. Controls received the inactive enantiomer N(omega)-nitro-D arginine methyl ester (D-NAME). Regeneration was assessed by serial recordings of the M potential from interosseous muscles of the foot innervated by sciatic tibial motor fibers and morphometric analysis of myelinated fibers distal to the injury site. Contrary to expectation, M potentials reappeared earlier in the mice treated with L-NAME and were higher in amplitude (reflecting the number of reinnervating motor fibers) at 10 weeks after the injury. In the L-NAME treated mice, the mean axonal diameter of regenerating tibial myelinated fibers was larger and the fiber size histogram was shifted to larger fibers. Inhibition of NOS in a transected peripheral nerve is associated with enhanced regeneration of myelinated fibers. Local elaboration of NO may be toxic to regenerating axons. PMID- 9209102 TI - Human stance stability improves with the repetition of the task: effect of foot position and visual condition. AB - The effects of repetition of quiet stance trials on body sway, recorded through a stabilometric platform, were studied in 12 normal subjects. With feet together, both with eyes open (EO) and closed (EC), a progressive shift forward of the centre of foot pressure (CFP) occurred with repetition. In addition, with EC, but not with EO, a significant progressive reduction in sway area (SA) and sway path (SP) occurred. With feet 10 cm apart, initial SA and SP values were significantly smaller than with feet together, regardless of the visual condition, but repetition of trials induced no significant effects on either position of CFP or body sway under either visual condition. Results indicate the occurrence of a learning phenomenon in this simple postural task, whereby the body shifts towards a 'safer' position with a minimum energy expenditure due to reduced corrections of sway. Forward leaning and decrease in sway are two independently-occurring processes, each possibly due to a better central integration of proprioceptive input with repetition of trials. PMID- 9209103 TI - Cortical 5-HT-CCK interactions and anxiety-related behaviour of guinea-pigs: a microdialysis study. AB - Serotonin (5-HT) and cholecystokinin (CCK) are involved in the development of anxiety. There are only few data suggesting interactions between CCK and 5-HT under aversive conditions. In our study the cholecystokinin tetrapeptide (CCK-4) (10 microg/kg) induced 'anxious' behaviour and potentiated the increase of 5-HT release on the elevated plus maze (X-maze). The 'anxiolytic' 5-HT1A agonist 8 hydroxy-2-(di-n-propyl amino) tetralin (8-OH-DPAT; 0.3 mg/kg) reduced basal 5-HT and the increase in 5-HT release on the X-maze. 8-OH-DPAT given simultaneously with CCK-4, blocked the effects of CCK-4. The results demonstrate an interaction between CCK and 5-HT1A mechanisms via the influence on cortical 5-HT release. PMID- 9209104 TI - Axonal branching and termination of cervical reticulospinal neurons in the cat brachial segments. AB - Axonal branching patterns in the brachial segments of cervical reticulospinal neurons (C-RSNs) were examined in cats using intraaxonal injection of horseradish peroxidase (HRP). Axons of these neurons were electrophysiologically identified by their projection to the lower cervical but not to the lumbar segments and monosynaptic activation after tectal and pyramidal stimulation. Six axons were stained up to terminals. Their stem axons descended in the ventral funiculus near the boundary of the spinal gray. The majority of collateral axons crossed lamina VIII and distributed terminals in the whole area of lamina VIII, the middle part of lamina VII, the lateral 2/3 of lamina VI, the ventral part of lamina V, and sparsely in the limb motor nuclei. Their possible functional role in head orienting movements is discussed. PMID- 9209105 TI - Intracortical inhibition and facilitation are abnormal in Huntington's disease: a paired magnetic stimulation study. AB - Transcranial magnetic stimulation with a conditioning-test paradigm was used to assess cortico-cortical interactions in the motor cortex of 11 patients with Huntington's disease (HD) as compared to normal controls (NC). In the HD patients, threshold and amplitude of motor potentials evoked in the opponens pollicis muscle at rest were not significantly different from NC. The cortico cortical inhibition at interstimulus intervals of 2-5 ms was significantly reduced and the cortico-cortical facilitation at longer intervals (10-25 ms) was significantly enhanced. Changes of intracortical inhibition and facilitation were related to clinical rating of choreic dyskinesias. The data support the hypothesis of a functional impairment of the motor cortex-basal ganglia loop in HD patients. PMID- 9209106 TI - Neuroligand-triggered calcium signalling in cultured human glioma cells. AB - Cells from primary cultures of four glioblastomas (GB), three low-grade astrocytomas (A), and four low-grade oligodendrogliomas (O) were tested for the presence of neuroligand receptors linked to Ca2+ signalling by calcium imaging. Cells of days 3 to 21 in culture were incubated with 5 microM fluo-3 acetomethylester in a bath solution and stimulated with 0.1 mM ATP, 0.01 mM angiotensin II, bradykinin, histamine, norepinephrine, serotonin, and substance P for 15 s, with 0.01 mM glutamate and 50 mM K+ for 30 s. Changes in the Ca2+ concentration were measured with a confocal laser scanning microscope. In all glioma subtypes, the majority of cells showed Ca2+ responses after application of histamine (60% of cells tested in GB, 67% in A, 86% in O), bradykinin (66% in GB, 29% in A, 55% in O) and ATP (48% in GB, 70% in A, 47% in O). The other stimuli induced Ca2+ transients in a smaller proportion (between 33% and 2%) of the cells. Our study demonstrates that histamine, bradykinin and ATP are potent inducers of [Ca2+]i signals in gliomas. PMID- 9209108 TI - Apoptosis induced in the spinal cord and dorsal root ganglion by infection of herpes simplex virus type 2 in the mouse. AB - After inoculation of a highly neuro-invasive strain of herpes simplex virus (HSV) type 2 (186) into the mouse hind-paw planter skin, many virus-positive neurons and glial cells were detected in the dorsal root ganglia (DRGs) and lumbar spinal cord by immunohistochemistry for HSV-2 antigen. A number of apoptotic cells were also observed in the spinal cord and DRGs by the terminal dUTP nick-end-labeling (TUNEL) method. Double labeling with the immunohistochemistry for HSV-2 and the TUNEL method revealed further that some glial and neuronal cells in the spinal cord, either infected or non-infected by HSV-2, showed apoptotic signs. In DRGs, however, apoptosis was detected in no neuronal cells, although some of HSV-2 infected glial cells were apoptotic. PMID- 9209107 TI - Characteristics of mucosal nociceptors in the rat oral cavity: an in vitro study. AB - Characteristics of mucosal nociceptors were investigated by recording activities from single fibers in the lingual nerve in an in vitro jaw-nerve preparation of rats. We found four subtypes of nociceptors in the medial side gingival area of the oral mucosa in the lower jaw: 11 A delta-high threshold mechanonociceptors (A delta-HTMs), 7 A delta-mechanoheat nociceptors (A delta-MHs), 21 A delta polymodal nociceptors (A delta-POLYs) and 28 C-polymodal nociceptors (C-POLYs). Thus the majority of the nociceptors was polymodal type (ca. 73% of the recorded fibers), ca. 43% of which consisted of A delta-POLYs scarcely found in the skin but commonly found in the deep tissue, muscle or colon. In contrast to the skin, the size of the receptive field of both A delta and C polymodal types was larger than that of either A delta-HTM or A delta-MH type. The von Frey threshold of all types of mucosal nociceptor was higher than that of the skin, though their heat threshold was almost the same as that of the skin nociceptors. These results show that the mucosal nociceptors are different from the skin nociceptors in the frequency distribution of their types and certain physiological properties. PMID- 9209109 TI - L-aspartate-evoked inhibition of melatonin production in rat pineal glands. AB - Our previous studies in rat indicated that pinealocytes secrete L-glutamate through microvesicle-mediated exocytosis to regulate negatively melatonin production. Recently, we further found that pinealocytes secrete L-aspartate through microvesicle-mediated exocytosis. In the present study, we investigated the role of L-aspartate in the melatonin production in isolated rat pineal glands. It was found that L-aspartate inhibits norepinephrine-stimulated melatonin production as well as serotonin N-acetyltransferase activity reversibly and dose-dependently, the concentrations required for 50% inhibition being 150 and 175 microM, respectively. L-Asparagine and oxaloacetate, metabolites of L aspartate, had no effect on the melatonin production. These results suggest that pinealocytes use L-aspartate, as well as L-glutamate, as a negative regulator for melatonin production. PMID- 9209111 TI - Protein kinase C in the postmortem brain of teenage suicide victims. AB - Increased serotonin2A (5-HT2A) receptors have been reported in the postmortem brain of suicide victims. To examine if this increase is associated with the dysregulation of postreceptor sites in the signaling cascade, we determined [3H]phorbol dibutyrate (PDBU) binding to protein kinase C (PKC) in postmortem brain samples (Brodmann's areas 8 and 9) obtained from teenage suicide victims and control subjects. [3H]PDBU binding to PKC was determined in membranal and cytosolic fractions. We observed that Bmax of [3H]PDBU binding sites was significantly decreased in both membranal and cytosolic fractions in brain samples from Brodmann's areas 8-9 compared to matched controls. These results thus suggest that PKC may play a role in the pathophysiology of suicidal behavior. PMID- 9209110 TI - Nicotinamide inhibits inducible nitric oxide synthase mRNA in primary rat glial cells. AB - Nitric oxide (NO) exerts cytotoxic effects on various cells including neuronal cells. Glial NO production, mediated via induction of inducible NO synthase (iNOS), enhances neurotoxicity associated with the N-methyl-D-aspartate (NMDA) receptor. The present study examined whether nicotinamide, an inhibitor of poly (ADP-ribose) synthetase, inhibits NO formation in primary culture of rat glial cells. Nicotinamide (5-20 mM) suppressed iNOS mRNA expression and subsequent NO formation, which were induced by the combination of interferon-gamma and lipopolysaccharide, in a dose dependent manner. In addition, high-concentration (20 mM) nicotinamide decreased mRNA of interferon regulatory factor-1, a transcription factor which plays a major role in iNOS mRNA induction. These results suggest that nicotinamide may have protective effect on glial NO-related pathologies by preventing iNOS mRNA induction. PMID- 9209112 TI - Glutamate-mediated activation of protein kinase C in hippocampal neurons. AB - Exposure of cultured neurons to glutamate results in the activation of protein kinase C (PKC). However, the mechanisms of this activation process are incompletely understood. To investigate PKC activation in cultured rat hippocampal neurons, we have exposed these cells to 500 microM glutamate for 5 min prior to determination of PKC phosphotransferase activity and protein levels. We observe that PKC activity increases in the membrane fractions by 3.7-fold, but is unchanged in the cytosol. Protein levels are also significantly increased by 5.5-fold in the membrane fractions. These results indicate that in hippocampal neurons, activation of PKC by glutamate does not occur solely by the translocation of PKC from the cytosol to the membrane. We suggest that neurotoxic concentrations of glutamate activate PKC in a translocation-independent manner which may serve as a mediator for glutamate-induced neurotoxicity. PMID- 9209113 TI - Regionally specific effects of haloperidol and clozapine on dopamine reuptake in the striatum. AB - This study examined the extent to which local application of the typical neuroleptic haloperidol (HAL) or the atypical neuroleptic clozapine (CLOZ) influences dopamine (DA) transporter function in the dorsal and ventral striatum. Using urethane-anesthetized rats, DA was pressure ejected and monitored with in vivo electrochemistry, into the dorsal and ventral striatum to establish regional baseline DA reuptake rates. Haloperidol or CLOZ (10 microM) was then applied, followed 5 min later by DA, in order to assess drug effects on DA reuptake rates. Haloperidol caused a 62% decrease in dorsal striatal DA reuptake rates while CLOZ had no effect on reuptake rates. Neither neuroleptic significantly altered DA reuptake rates in the ventral striatum. It is possible that HAL-induced decrease in DA reuptake in the sensorimotor striatum could be related to the motor side effect profile of this neuroleptic. Additional studies with other typical and atypical neuroleptics are needed to further evaluate the relationship between slowing of DA reuptake and the side effect potential of neuroleptic agents. PMID- 9209114 TI - Activity-dependent survival of rat cerebellar granule neurons is not associated with sustained elevation of intracellular Ca2+. AB - Ca2+ plays a pivotal role for the activity-dependent survival of neurons. In primary culture of cerebellar granule neurons, we found that there is no significant difference in intracellular Ca2+ level in the survival-promoting condition (cultures in the presence of 25 mM KCl) and that in the apoptosis inducing condition (cultures in the presence of 5 mM KCl). This was not due to the inactivation of voltage-dependent L-type Ca2+ channels in the survival promoting condition, but due to the enhanced rate of the influx and the efflux of Ca2+ in the survival-promoting condition compared to that in the apoptosis inducing condition. These results suggest that the activity-dependent survival of the granule neurons is not associated with sustained rise of intracellular Ca2+ but associated with the enhanced turnover rate of Ca2+. PMID- 9209115 TI - Role of histidyl residues in the binding of ligands to the porcine N-methyl-D aspartate receptor. AB - Possible involvement of histidyl residues in the binding of ligands to ionotropic glutamate receptors and to modulatory sites on the N-methyl-D-aspartate (NMDA) receptor was assessed in porcine cortical synaptic plasma membranes after covalent modification with diethyl pyrocarbonate (DEPC). Binding of [3H]glutamate to the NMDA sites was enhanced but to the 2-amino-3-hydroxy-5-methyl-4 isoxazolepropionate (AMPA) and kainate receptors unaffected by 1 and 5 mM DEPC. Binding of 3-[(R)-carboxypiperazin-4-yl]-[1,2-(3)H]propyl-1-phosphonate ([3H]CPP) was reduced in a dose-dependent manner by DEPC and the activation of binding by 1 hydroxy-3-amino-2-pyrrolidone (HA-966) blocked by 10 mM DEPC. DEPC reduced the strychnine-insensitive binding of [3H]glycine and the glycine- and glutamate activated binding of [3H]dizocilpine. Protection experiments indicated that histidyl residues are directly involved in the binding of glycine (but not HA 966) and allosterically modulate the binding of glutamate, CPP and dizocilpine. The results corroborate the existence of agonist- and antagonist-preferring sites or conformational states of the NMDA receptors. PMID- 9209116 TI - Circadian rhythms in vitamin B12 content of the rat brain. AB - The whole brain content of vitamin B12 (VB12) in the rat was assayed by the chemiluminescent immunoassay at 4 h intervals in 12:12 h light-dark cycles (LD) and five different circadian times (CTs) under constant dim illumination (dim LL). In LD-entrained rats, the content of VB12 exhibited a day-night rhythm with a peak 2 h after the dark onset time and a trough 2 h before the light onset time. In freerunning rats under dim LL, the content of VB12 also exhibited a circadian variation with a peak ca. 2 h after the activity onset time (CT14) and a trough ca. 12 h after the activity onset time (CT 0). These findings clearly indicate that the brain VB12 content decreased during the active phase and increased during the resting phase regardless of the lighting schedule. On the other hand, the drinking behavior, as an index of the intake activity, was observed less frequently in the light phase of LD cycles and the resting phase of dim LL. Since most of the digested VB12 is known to be continuously stored in the liver, it is demonstrated that the rhythm of the brain content of VB12 may be caused by brain consumption of VB12 during the active phase and the transportation from the peripheral storage during the resting phase, independent of intake activities. PMID- 9209117 TI - Coexistence of calcitonin gene-related peptide and NADPH-diaphorase in the canine superior cervical ganglion. AB - By means of double staining technique of NADPH-diaphorase (NADPH-d) histochemistry and calcitonin gene-related peptide (CGRP) immunohistochemistry, we investigated the coexistence of NADPH-d reactivity and CGRP immunoreactivity in the canine superior cervical ganglion (SCG). Most of NADPH-d reactivity and CGRP immunoreactivity were coexisted in the principal postganglionic neurons. These neurons were distributed throughout the ganglion without specific localization. The present findings suggest the intimate role of CGRP and nitric oxide in postganglionic neurons of the canine SCG. PMID- 9209118 TI - Developmental changes of inward rectifier currents in neurons of the rat entorhinal cortex. AB - A slice preparation was used to investigate inward rectifier currents (I(H)) of entorhinal cortex (EC) neurons. Using the whole-cell configuration of the patch clamp technique, I(H) was studied in pyramidal cells from layer IV of the rat EC and in stellate cells from layer II of the EC. Inward rectifier currents were analyzed in neurons of newborn (P1-3), juvenile (P8-14) and adult (>P28) rats. Pyramidal cells of juvenile rats possessed a significantly larger current density of I(H) than pyramidal cells of newborn rats, whereas no differences in the current density of I(H) were found between pyramidal neurons of juvenile and of adult animals. In contrast, the current density of I(H) of stellate cells was significantly increased in juvenile rats compared with newborn rats as well as in adult rats compared with juvenile rats. Moreover, in adult rats the current density of I(H) was larger in stellate cells than in pyramidal cells, whereas opposite data were obtained in juvenile animals. PMID- 9209119 TI - [Study of bacterial flora in the oral cavity and stomach of elderly patients receiving nasogastric tube feeding]. AB - To investigate the significance of oropharyngeal flora and gastric flora in elderly patients receiving nasogastric tube feeding, throat secretions and gastric aspirates were cultured and the pH of the latter was measured. Of 116 bacterial isolates from throat secretions of 27 elderly patients, 30 were beta streptococci and 28 were Pseudomonas aeruginosa. Bacteria isolated from gastric aspirates numbered 86 and 24 (27.9%) of them were the same species as those found in the throat secretions. Patients with gastric pH were below 3.5 had significantly lower concentrations of gram-negative bacili in gastric aspirates. We also studied oropharyngeal flora in 33 elderly patients who were admitted to Nagoyashi Koseiin Geriatric Hospital. The major bacterial isolates from throat swabs of bedridden patients were gram-negative bacilli and beta-streptococci, especially group B streptococci (GBS). We measured the level of antibody to GBS in these patients. Those from whom GBS were isolated had high titers. These results suggest that in elderly patients receiving enteral nasogastric) tube feeding, large numbers of bacteria colonize the oral cavity and stomach. The measurement of type-specific antibody to GBS may be useful in managing such patients. PMID- 9209120 TI - Duration of chronic HCV infection and efficacy of interferon in chronic hepatitis C patients with a history of blood transfusion. The Study Group for Treatment of Hepatitis in the Kyushu Area. AB - To investigate correlations between the interval between blood transfusion and the start of IFN therapy, and IFN efficacy, we studied chronic hepatitis C patients with a history of blood transfusion. The subjects were 122 patients with chronic hepatitis C and a history of blood transfusion at 64 institutions. The patients were treated with high or low-dose IFN. High-dose therapy consisted of intramuscular injection of human lymphoblastoid interferon (HLBI), 6 x 10(6) IU daily for 2 weeks, then 3 times a week for 22 weeks, and low-dose interferon therapy of intramuscular injection of HLBI, 6 x 10(6) IU daily for 2 weeks, then 3 x 10(6) IU 3 times a week for 22 weeks. Normal serum ALT levels for 6 months or more after completing IFN (complete response) were found in 44/122 (36.2%) patients and HCV RNA was no longer detectable after completing IFN therapy in 19/68 (27.9%). Patients in whom the interval between blood transfusion and the start of IFN therapy was less than 20 years had significantly higher rates of HCV RNA-negative complete response than those in whom the interval was 20 years or more (p < 0.039). When chronic HCV infection is caused by blood transfusion, the efficacy of IFN depends on the duration of chronic HCV infection. Since the duration of HCV infection is a factor in predicting efficacy, early IFN therapy may be more effective. PMID- 9209121 TI - [Shigella dysenteriae strains possessing a new serovar isolated from imported diarrheal cases in Japan]. AB - Two Shigella strains (93-119 and 95-619) isolated from stool cultures of imported diarrheal cases in Japan, did not react to any antisera of the established SHigella serovars. These strains had the typical biochemical characteristics of Shigella dysenteriae, and were biochemically identical to each other. Both strains were positive in the Sereny test and other tests for invasiveness; these indicate that they can cause shigellosis in humans. The results of antigenic analysis showed that they did not belong to any of the recognized or provisional serovars, and were serologically indistinguishable. Strain 93-119 is designated as the test strain for this new serovar. PMID- 9209123 TI - [Prevalence of antimicrobial resistance among clinical isolates of Streptococcus pneumoniae in a children's hospital]. AB - Eleven hundreds and seventy-six strains of Streptococcus pneumoniae were isolated from pediatric clinics of Chiba Children's Hospital during 1990 through 1995. Annual penicillin-resistant rates of these strains were as follows; 24.0% (1990), 29.0% (1991), 36.2% (1992), 55.8% (1993), 58.6% (1994), and 59.3% (1995). Overall penicillin-resistance during these 6 years was 45.8%. Nine out of 11 cases of systemic pneumococcal infections were due to penicillin-resistant S. pneumoniae (PRSP) during the same period. Of PRSP strains, their PCG-MIC levels had become higher and their spectra of resistance had expanded not only to beta-lactam but also to non-beta-lactam antimicrobials. Although panipenem was the most efficacious antibiotics in this study and was recommended currently to use in the case of pneumococcal meningitis, it should be noted that a strain with high-level MIC (2 micrograms/ml) had emerged in 1995. Close surveillance of pneumococcal antimicrobial susceptibility including panipenem is necessary. PMID- 9209122 TI - [Incidence of Kanagawa phenomenon-positive and -negative Vibrio parahaemolyticus strains isolated from traveller's diarrhea and their relation to tdh and trh genes]. AB - A total of 1,319 strains of Vibrio parahaemolyticus isolated from traveller's diarrhea were analysed for Kanagawa phenomenon (KP) with the Wagatsuma blood agar test and the results were also compared with those of analyses of tdh and trh genes which encode thermostable direct hemolysin (tdh) and TDH-related hemolysin (trh). The majority of the strains (1,152 strains) counting 87.3% had positive KP, among which 1,049 and 103 strains were only tdh and both tdh and trh-positive ones, respectively. However, 167 strains counting 12.7%, which is quite high compared to the previous report, were found to have negative KP, among which 94 and 24 strains were only trh and both tdh and trh-positive ones, respectively. PMID- 9209124 TI - [The evaluation of interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF alpha) level in peripheral blood of patients with chronic lower respiratory tract infection]. AB - In the present study, we assessed the serum level of IL-6 and TNF-alpha by ELISA in patients with chronic lower respiratory tract infection. The serum levels of IL-6 and TNF-alpha of patients in acute exacerbation phase are higher than that of in stable phase. We also classified patients in acute exacerbation phase into two groups according to the microorganism of persistent infection. The serum level of IL-6 and TNF-alpha in the patients with persistent infection with Pseudomonas aeruginosa were higher than that with Haemophilus influenzae. Moreover, the serum level of IL-6 and TNF-alpha were found to be related with malnutrition which assessed by clinical indices such as the serum level of albumin and cholinesterase. The present result suggests that IL-6 and TNF-alpha may have relationship with not only inflammation in airway but also indices of nutrition in patients with chronic lower respiratory tract infection. PMID- 9209125 TI - [Suppressive effect of clarithromycin on the production of verotoxin by E. coli O157]. AB - Effects of low concentration of clarithromycin (CAM) on the production of Verotoxin (VT) by Escherichia coli O157 was investigated in vitro. The production of VT1 was suppressed up to 10 hours when bacteria was incubated with 0.64 micrograms/ml (equivalent to the 1/100th of MIC) or higher concentrations. However, the production of VT1 reached to the control level after 22 hours even with 6.4 micrograms/ml of CAM. On the other hand, production of VT2 by 22 hours was partially suppressed with 0.64 micrograms/ml of CAM and completely with 6.4 micrograms/ml of CAM. When the eight clinical isolates were incubated with 0.64 or 6.4 micrograms/ml of CAM, VT1 and VT2 were suppressed in two and eight strains, respectively. In these strains, similar but less efficient suppression of the toxin production was also observed with erythromycin. In contrast to the macrorides, ampicillin did not inhibit or rather stimulated the production of VT1 and VT2 in some strains. PMID- 9209126 TI - [Antibacterial actin of vinegar against food-borne pathogenic bacteria including Escherichia coli O157:H7 (Part 1). Examination of bacteriostatic and bactericidal activities]. AB - Bacteriostatic and bactericidal activities of vinegar products against food-borne pathogenic bacteria including enterohemorrhagic Escherichia coli (EHEC) O157:H7, and other bacteria were examined. By the presence of spirit vinegar at 0.1% acidity, the growth of 34 strains of bacteria was completely inhibited. Grain and rice vinegars also inhibited the growth of bacteria at the same acidity. These results suggest that vinegars have strong bacteriostatic activity. Bactericidal activity of vinegar products was measured. The order of their activities against E. coli O157:H7 strains was spirit vinegar > grain vinegar > rice vinegar. Susceptibility of 7 EHEC strains (6 E. coli O157:H7 isolated from 3 outbreaks and 1 E. coli O26:H11 from a sporadic case) to spirit vinegar was similar to each other, and much lower than that of an enteropathogenic E. coli O111:K58:H- strain. It indicates that these EHEC strains are rather acid-tolerant. The bactericidal activities of vinegars were independent of bacterial inoculum sizes, but were dependent of growth phase. Bacteria of logarithmic growth phase were more sensitive than those of stationary phase. Bactericidal activity of spirit vinegar profoundly depended on the reaction temperature. At higher temperatures, spirit vinegar killed bacteria much more rapidly. In 2.5% acidity vinegar, the times for the log3 reduction of cells were 4,516 min at 10 degrees C, 739 min at 20 degrees C, 137 min at 30 degrees C, 14.4 min at 40 degrees C, and 0.84 min at 50 degrees C, respectively. These results suggest that treatment with vinegar solution at handy temperatures ranged 40-50 degrees C may be one of the useful methods to prevent bacterial food poisoning. PMID- 9209127 TI - [Antibacterial actin of vinegar against food-borne pathogenic bacteria including Escherichia coli O157:H7 (Part 2). Effect of sodium chloride and temperature on bactericidal activity]. AB - Bactericidal effects of various kinds of AWASEZU (processed vinegar, 2.5% acidity) on food-borne pathogenic bacteria including Escherichia coli O157:H7 and other bacteria were examined. the order of bactericidal activities was NIHAIZU (3.5% NaCl was added) > SANBA-IZU (3.5% NaCl and 10% sucrose were added) > plain vinegar (spirit vinegar) > AMAZU (10% sucrose was added). This indicates that their activities were enhanced by the addition of sodium chloride and suppressed by the addition of sugar. On the other hand, when soy sauce was used instead of sodium chloride, the order of bactericidal activities was plain vinegar > AMAZU > NIHAIZU > SANBAIZU. This is mainly because their activities were suppressed by the increase in the pH value. The effect of sodium chloride (0.01-15%) and temperature (10-50 degrees C) on bactericidal activities against E. coli O157:H7 in spirit vinegar (0.5-2.5% acidity) was further examined. When vinegar was used in combination with sodium chloride, predominant synergism on the bactericidal activity was observed. Their activities were markedly enhanced by the addition of sodium chloride in proportion to the concentration. In addition to this, at higher temperatures spirit vinegar killed bacteria much more rapidly. It should be noted that the bactericidal activity of spirit vinegar was extremely enhanced by the combined use of the addition of sodium chloride and the rise of temperature. For example, in 2.5% acidity vinegar, the time required for 3 log decrease in viable cell numbers at 20 degrees C was shortened to 1/140-fold by the addition of 5% sodium chloride, shortened to 1/51-fold by the rise of the reaction temperature at 40 degrees C, and shortened to 1/830-fold; 0.89 minutes by both the addition of 5% sodium chloride and the rise of temperature at 40 degrees C. In order to propose the methods to prevent food poisoning by bacterial infection, bactericidal activities of vinegar solution containing sodium chloride on cooking tools and raw vegetables were examined. Vinegar solution (1-2% acidity, 3-7% NaCl) produced more than 3 log decrease in viable cell numbers of E. coli O157:H7 on the surface of cutting board, and cabbage and cucumber at 20 50 degrees C. These results suggested that the treatment with vinegar solution containing sodium chloride may be one of the useful methods to prevent food poisoning. PMID- 9209128 TI - [A case of severe odontogenic infection with allergic skin reaction for beta lactam antibiotics]. AB - A 54-aged woman consulted with right buccal phlegmon caused by lower apical periodontitis. The primary chemotherapy using flomoxef induced allergic skin eruption. A decision was made for using clindamycin, levofloxacin and fosfomycin as chemotherapeutic agents. Cultured organisms from the abscess revealed the mixed infection with Streptococcus sanguis, Veillonella sp., Prevotella loescheii, Wolinella spp. On 12th hospital day, her serum CRP turned to negative. This case carried numerous implications for the use of antibiotics as chemotherapy agents. PMID- 9209129 TI - [A case of Tsutsugamushi disease as an imported infection]. AB - Tsutsugamushi disease is widely spread throughout Japan. A case of tsutsugamushi disease was seen in October, 1996. A 64-year-old male developed typical symptoms of tsutsugamushi disease with Rickettsia tsutsugamushi, after he returned to Japan from Cheju Island, Korea. Not only in Japan but also in other Asian countries including Korea, China, Taiwan, and Thailand, tsutsugamushi disease is one of the most important rickettsial diseases carried by ticks or mites. If a traveller returning from an Asian country has symptoms such as high fever, skin eruption, and lymphadenitis, we should suspect that he is suffering from tsutsugamushi disease and should search if he has an eschar on any area of his body. We should not forget that tsutsugamushi disease is an imported disease. Patients of tsutsugamushi disease often have hematological disorders. They are sometimes referred to the hematological section of the hospital. Hematologists should be familiar with this disease. PMID- 9209130 TI - [A girl with systemic onset juvenile rheumatoid arthritis suspected to be due to human parvovirus B19 infection]. AB - We encountered a case of a girl where Human Parvovirus B19 infection was considered to have been concerned with the development of systemic type juvenile rheumatoid arthritis (JRA). While the affected child did not show any evident infectious erythema-like findings, changes in the serum antibody titer by the EIA method presented the pattern of first infection. During the clinical course the condition of the disease as JRA was serious and hemophagocytic syndrome developed concurrently. Furthermore, the resistance to the treatment was also noted. So the patient was treated with prednisolone combined with low dose weekly MTX therapy. The possibility of Human parvovirus B19 being concerned with the development of rheumatoid arthritis in one form or another has been suggested in recent years. In the disease type with systemic angititis as main pathophysiology, which is called systemic JRA we encountered this time, it is not clear how Human Parvovirus B19 was concerned with the development of this disease, but it appeared to hold a key position in studying pathophysiology of the development. PMID- 9209131 TI - [The patients without specific antibodies, were diagnosed by PCR as Tsutsugamushi disease]. PMID- 9209132 TI - [Inhibition of corneal neovascularization by a new analog of fungal product]. AB - The anti-angiogenic activity if FR 118487, a new synthetic analogue of Scolecobasidium arenarium products, was examined in corneas of Japanese white rabbits. We studied locally administered FR 118487 with hydron pellets in two models; corneal neovascularization after implantation of a CuCl2 pellet (CuCl2 induced model) and a Wistar rat's cornea (keratoplasty model) into a rabbit cornea. In the CuCl2-induced model, maximum length of neovascularization was 0.08 +/- 0.10 (mean +/- standard deviation) mm with FR 118487 (control 2.84 +/- 0.13 mm) at 1 week after the implantation. In the keratoplasty model, maximum length of neovascularization was 0.05 +/- 0.05 mm with FR 118487 (control 3.33 +/- 0.18 mm) at 2 weeks after the implantation. In both models, FR 118487 had a significant (p < 0.01) effect on inhibition of corneal neovascularization. PMID- 9209134 TI - [Choroidal lesions and surgical treatment of bullous retinal detachment]. AB - To define choroidal lesions of bullous retinal detachment, we performed indocyanine green angiography (IA) on 20 eyes of 10 patients with bullous retinal detachment. Four of these eyes were treated with sclerectomy and sclerostomy (Gass), and the benefit was evaluated. IA showed the following choroidal abnormalities: choroidal filling delay in the macular region (6 of 18 eyes, 33%) and underneath the leaking sites (9 of 18 eyes, 50%), localized (1 eye, 5%) and extended (18 eyes, 90%) choroidal venous dilatation, and intrachoroidal hyperfluorescence (all eyes, 100%). After the surgical treatment, retinal detachments resolved within 9 weeks (mean 5.3 weeks). Since choroidal filling delay, choroidal venous dilatation and intrachoroidal hyperfluorescence were observed, it is suggested that choroidal congestion plays a causative role in bullous retinal detachment. Sclerectomy and sclerostomy seemed to be beneficial in treatment of this condition. PMID- 9209133 TI - [Ornithine induced retinopathy in rat--2. Process of disappearance of choriocapillaris]. AB - We observed the process of disappearance of the choriocapillaris after loss of the retinal pigment epithelium (RPE) induced by intravitreal injection or ornithine. Three hours after administration of ornithine, the RPE cells swelled remarkably in the posterior pole, but, the endothelial cells of the choriocapillaris remained intact. At 3 days, the RPE cells became necrotic, but the choriocapillaris still preserved its in normal appearance. At 7 days, RPE disappeared completely in the posterior pole and the choriocapillaris displayed evidence of atrophy; the swollen lumen of the choriocapillaris became narrow the cytoplasm of the endothelium was swollen, and the number of fenestrae was reduced. On the other hand, these changes were not seen where the RPE remained. At 14 days, in the posterior pole, the lumen of the choriocapillaris occluded by the swollen endothelial cells. At 28 days, the choriocapillaris completely disappeared and the large choroidal vessel was directly in contact with Bruch's membrane. These results showed that the RPE is correlated with the presence of the choriocapillaris. PMID- 9209136 TI - [Geometrical distribution of newly formed blood vessels in diabetic retinopathy]. AB - Outbreak sites of newly formed blood vessels (NFVs) in proliferative diabetic retinopathy (PDR) were analyzed geometrically. A montage was made of fluorescein fundus angiography photographs of 109 eyes and applied to a two dimensional x-y plane in which the center of the optic disc was taken as the origin. Thus each NFV acquired a coordinate value (x, y). After a coordinate transformation of this x-y plane designed to correct the parafoveal distortion of vascular distribution, a polar coordinate value (r, theta) of the NFV was calculated. The frequencies of NFVs were analyzed statistically relating to both theta, which is the angle around the origin, and r, which is the distance from the origin. The average theta was 2.74 +/- 1.72 (mean +/- standard deviation) radians and the outbreak frequency of NFVs showed a homogeneous distribution which was not influenced by any difference in theta. As for r, the frequency corrected as a truncated distribution at r = 0 showed a normal distribution with an average of 1.45 +/- 0.8. The outbreak of NFVs in PDR is a phenomenon in which accidental elements are strongly concerned and it is thought to be difficult to predict the site at early stage of retinopathy. PMID- 9209135 TI - [Quantitative evaluation of aqueous flare in psoriasis using a laser flare-cell meter]. AB - We evaluated aqueous humor protein concentration in psoriasis using a laser flare cell meter, which can quantify aqueous flare precisely and objectively. Psoriatic severity was evaluated on the basis of psoriasis area and severity index (PASI) score. Aqueous flare was measured in 40 eyes of 20 psoriasis patients (sixteen psoriasis vulgaris, three guttate psoriasis, and one psoriatic arthritis) and 28 eyes of 14 normal controls. Aqueous flare value was significantly higher in psoriatic patients than in normal controls (p < 0.01). There was no difference between psoriasis vulgaris and the other types of psoriasis. Aqueous flare value was higher in patients with psoriatic history longer than 10 years than in those with less than 10 years (p < 0.05), and also higher in patients with severe psoriasis (PASI score > 10) than in those with mild psoriasis (PASI score < 10) (p < 0.05). But no statistically significant differences in aqueous flare value were found among cyclosporin, etretinate, and psoralen ultra violet A therapies. These findings strongly suggest that patients suffering from psoriasis have slight damage of the blood-aqueous barrier even if they have no ocular symptoms, and that the degree of blood-aqueous barrier damage increases with time and severity of psoriasis. PMID- 9209137 TI - [Concentration of vascular endothelial growth factor within the subretinal space and vitreous fluid in rhegmatogenous retinal detachment]. AB - To determine the increased production and release of vascular endothelial growth factor (VEGF) from the retina in the eye with non-angiogenic retinal detachment in which relative blood supply disturbance may be present, the concentration of VEGF in subretinal fluid and vitreous fluid collected from the eyes was investigated by enzyme linked immunospecific assay. The average concentration of VEGF was 0.5 +/- 1.1 ng/ml (mean +/- standard deviation) in nine samples of vitreous fluid collected from proliferative retinal detachment, and was 1.2 +/- 1.2 ng/ml in eleven samples of subretinal fluid from rhegmatogenous retinal detachment. The concentration of VEGF in six samples of vitreous fluid from angiogenic diabetic eyes (5.0 +/- 2.8 ng/ml) was significantly higher than in the samples from eyes with retinal detachment. The results suggest that the relative ischemic insult to the detached retina increases the release of VEGF into the vitreous cavity and subretinal space. The increased concentration of VEGF does not induce remarkable angiogenesis since the concentration is lower than the biological threshold, or the effect is modulated by inhibitors. PMID- 9209138 TI - [Factors affecting long-term visual outcome in retinopathy of prematurity treated with xenon photocoagulation and/or cryocautery]. AB - We evaluated long-term visual outcome in 28 patients (52 eyes) with retinopathy of prematurity after Xenon photocoagulation and/or cryocautery. The visual outcome was roughly classified into two groups: visual acuity of 0.6 or better and that of 0.2 or worse. The poor visual outcome resulted from macular degeneration, and risk factors for its development were small birth weight (Mann Whitney's U test, p = 0.03), retinopathy plus disease (chi 2 test, p = 0.041), treatment of a large area of the fundus (Mann-Whitney's U test, p = 0.035) and the inside of the vascular arcade (Mann-Whitney's U test, p = 0.0034), and treatment by cryocautery (chi 2 test, p = 0.032). Macular degeneration occurred either as an isolated small focus extending circumferentially around the fovea or as a result of extension from the temporal degeneration caused by photocoagulation. These results suggest that intensive treatment for retinopathy of prematurity, in addition to the prematurity by itself, caused the development of macular degeneration. Overtreatment should be carefully avoided in retinopathy of prematurity complicated by disease in which photocoagulation needs to be done in the area posterior to the ridge. PMID- 9209139 TI - [Immunohistochemical and immunogenetic analysis of ocular adnexal lymphoid proliferations]. AB - We examined 20 cases (21 specimens) of ocular adnexal lymphoid proliferations, using the histological, immunohistochemical and molecular genetic methods. The latter two protocols were performed to detect the light chains restriction of immunoglobulin with peroxidase-antiperoxidase (PAP) methods, and the clonality of immunoglobulin heavy chain gene with the hemi-nested polymerase chain reaction (PCR) method, respectively. Although in 8 cases it could not be morphologically determined whether they were neoplastic or not, clonality was revealed in 1 case with immunohistochemistry and in 4 cases with PCR. Two cases showed discordant results between immunohistochemistry and PCR probably due to somatic mutation of the framework region of the immunoglobulin heavy chain gene. Therefore, we, concluded that examination with these methods contribute to a better understanding of the nature of the ocular adnexal lymphoid proliferations. Furthermore, the immunoglobulin gene PCR method is very useful in practical examination, as it can be used with formalin-fixed paraffin-embedded specimens. PMID- 9209140 TI - [Surgical revision of failed filtering bleb with mitomycin C]. AB - The revision of a failed filtering bleb using mitomycin C was performed in 40 eyes (28 eyes with primary open angle glaucoma and 12 eyes with secondary open angle glaucoma) in which the previous trabeculectomy using 5-fluorouracil had failed to control intraocular pressure (IOP). Presurgical IOP ranged from 21 approximately 36 mm Hg (average: 24.7 mmHg) with the maximum tolerable antiglaucoma medication. The follow-up period was at least 2 years (24 approximately 60, mean: 39.8 months). Mitomycin C (0.04%, 5 ml, 5 min) was applied under the subconjunctival space. After rinsing, the wall of the scarred bleb was incised in cases with a localized bleb, or the margin of the scleral flap was incised in cases without a filtering bleb. The conjunctival wound was closed with 10-0 Nylon sutures. IOP control under 21 mmHg was achieved in 53.4% without medication and in 60.2% with or without medication at 60 months. After repeating the same procedure, IOP control was obtained in 82.2% with or without medication at 60 months. The success rate in secondary open angle glaucoma was less than in primary open angle glaucoma. Complications such as flattening of the anterior chamber requiring reformation (8%), choroidal detachment (8%), hyphema (5%), and hypotonic maculopathy (5%) were less than after trabeculectomy but conjunctival wound separation (5%) occurred little more frequently. PMID- 9209141 TI - [Four cases of late infection following scleral buckling procedure]. AB - We report four cases with late infection following a scleral buckling procedure. These cases were a 44-year-old male, a 52-year-old female, a 59-year-old male, and a 60-year-old male. All of these cases developed conjunctival injection, chemosis, and mucopurulent discharge 7 to 15 years after the procedure. The symptoms did not improve despite months of medical therapy, suggesting scleral buckle infection, and finally the buckling materials were removed. Solid silicone was used in 2 cases, and a sponge was used in 2 cases, with encircling elements in 1 case, and without encircling elements in 3 cases. Pus culture of the removal exoplant revealed fungaul infection in 2 cases. The others were negative, but bacterial infection was suspected from their clinical course and intraoperative findings. After removal of the buckles, the symptoms improved rapidly in all cases and there was no recurrence of retinal detachment. Scleral buckle infection should be suspected and removal of buckling materials should be considered, in cases with refractory conjunctival and scleral inflammation even years after scleral buckling procedure. PMID- 9209143 TI - [A case of sarcoidosis with proliferative retinopathy]. AB - A 19-year-old female manifested severe bilateral panuveitis with neovascularization in the iris, optic disc, and retina. Fluorescein fundus angiography showed dye leakage from the optic disc and retinal blood vessels, and a large non-perfused area was present in the peripheral retina of the right eye. Sarcoidosis was diagnosed histologically by conjunctival and skin biopsy. Although the patient was given a large dose of a corticosteroid systemically and received panretinal photocoagulation, a dense vitreous hemorrhage and cataract were apparent in the right eye. The right visual acuity decreased to hand motions. A pars plana lensectomy and vitrectomy were performed. After vitrectomy, inflammation and neovascularization regressed and the visual acuity improved to 20/100. Proliferative membrane obtained during vitrectomy was histopathologically studied by light and electron microscopy. Many new vessels containing neutrophils were observed. A direct effect of inflammation as well as ischemia in the retina may have been the stimulus for the proliferative changes. PMID- 9209142 TI - [Use of intravitreal ganciclovir for cytomegalovirus retinitis in a patient with systemic lupus erythematosus]. AB - A patient with systemic lupus erythematosus (SLE) developed bilateral cytomegalovirus (CMV) retinitis and was treated with a total of 9 intravitreal ganciclovir injections at a dose of 400 micrograms/50 microliters in each eye under topical anesthesia. She was unable to receive systemic antiviral therapy because of bone marrow suppression and renal failure. The condition of both eyes improved after the intravitreal injections. Recurrence of CMV retinitis was not seen until death. Intravitreal injections of ganciclovir are useful even for the non-acquired immunodeficiency syndrome (AIDS) immunocompromised patients with CMV retinitis when the patients can not receive intravenous medications because of systemic complications. PMID- 9209145 TI - The changing demographics of AIDS in Texas. PMID- 9209144 TI - [Human T-lymphotropic virus type 1 uveitis in children]. AB - We report here five pediatric patients with human T-lymphotropic virus type 1 (HTLV-I) uveitis. The patients were one boy and four girls aged between 3 and 14 years. The transmission route was considered to be breast feeding from their mothers. All patients had unilateral uveitis and the ocular symptoms were similar to those in HTLV-I uveitis in adults. The ocular inflammation responded to therapy with topical or systemic corticosteroids, but recurred in three patients. HTLV-I provirus DNA was detected by polymerase chain reaction (PCR) from infiltrating cells in the anterior chamber in one patient. The percentage of HTLV I-infected cells in the peripheral blood mononuclear cells was measured by quantitative PCR, and the values were high (2.9 approximately 7.3%) in three cases tested as compared with an asymptomatic carrier. These five cases show that HTLV-I uveitis can be induced in a relatively short period (3 approximately 10 years) after the viral infection, and that HTLV-I uveitis should be considered as one possible etiology of uveitis in children, particularly in a viral endemic area. PMID- 9209146 TI - Let doctors be doctors. Interview by Jean Pietrobono. PMID- 9209148 TI - Mediscare. PMID- 9209147 TI - Is health care only for the healthy?. Interview by Teri Moran. PMID- 9209149 TI - Attitude adjustment. PMID- 9209150 TI - Disease management. PMID- 9209151 TI - Minimizing the risk. PMID- 9209152 TI - Living with AIDS. PMID- 9209153 TI - Defusing the poor man's nuke. PMID- 9209154 TI - Discovering behavior. AB - The premise of this article is that animals behaving spontaneously sometimes show intriguing and unexpected behaviors. The theme of this article is that the animals producing these behaviors, when approached as psychological entities in their own right, offer unique information about how they perceive the world and carry out their lives. In addition, the findings often contribute to the modification, refinement, and generality of basic psychological concepts and principles. Many examples of such contributions are presented in recent reviews of psychological research with animals. In this article, the author illustrates a comparative approach to animal behavior with selected findings on emotional attachments in nonhuman primates. PMID- 9209155 TI - The analysis of Merino wool cuticle and cortical cells by Fourier transform Raman spectroscopy. AB - Wool fibers are comprised of proteins known as alpha-keratins and have a complex morphological structure. The major components of this structure, the cuticle and cortical cells, differ in the conformations of their chains as well as their amino acid compositions. High quality Fourier transform Raman spectra of cortical and cuticle cells isolated from fine Merino wool fibers have been obtained. Raman spectroscopy has been shown to be sensitive to the differences in both secondary structure and amino acid composition. The cortical cells were found to be higher in alpha-helical content as compared to the cuticle cells, which had an increased disordered content. Specific information, consistent with amino acid analysis results, regarding cystine, tyrosine, tryptophan, and phenylalnine residues, were obtained for both the cortical and cuticle cells. In addition, the Raman spectra provided information about free thiol groups, amino acids residues with amide group side chains, and residues with protonated carboxyl group side chains. Middle ir transmission spectra of these isolated cells were also obtained. In comparison to the Raman data, the middle ir spectra were found to be not as rich in information. PMID- 9209156 TI - Conformational study of asialo-GM1 (GA1) ganglioside. AB - In order to investigate the significance of preferred conformations of the saccharide for the steric orientation and recognition of glycosphingolipids at the membrane surface, the conformational free energy calculations were carried out on the asialo-GM1 [GA1; beta-D-Gal (1-->3) beta-D-GalNac(1-->4) beta-D-Gal(1- >4) beta-D-Glc-O-ceramide) using a new program CONCARB (CONformational study program for CARBohydrate) in the unhydrated and hydrated states. The overall backbone conformational of GA1 appears to be extended with a little bent at the glycosidic II-III linkage, in which two pyranose rings of Gal(IV)-GalNAc-(III) moiety orient approximately perpendicular to those of Gal(II)-Glc(I) moiety. This is consistent with the structures deduced from high-sensitivity differential scanning calorimetry experiments and the nmr study on GA1. The calculated glycosidic torsion angles of the lowest free energy conformation of GA1 in the hydrated state are in accord with the structures of relevant oligosaccharides deduced from nmr experiments and hard sphere exoanomeric calculations. A comparison of the values of glycosidic torsion angles phi and psi of GA1 and its constituent oligosaccharides indicates that the overall backbone conformation of each oligosaccharide is retained when the oligosaccharide chain becomes longer. This implies that the short-range interactions between the nearest-neighbored saccharides are of significant importance in stabilizing the overall backbone conformation of GA1 in both the unhydrated and hydrated states. The different orientation and hydrogen bonds of hydroxymethyl and hydroxyl groups from one oligosaccharide to another suggest that the medium- and long-range interactions are also of consequence. Hydration seems to affect significantly the confirmation of these groups, but not to perturb remarkably the overall backbone conformation of GA1. PMID- 9209158 TI - 1H-NMR analysis of CD3-epsilon reveals the presence of turn-helix structures around the ITAM motif in an otherwise random coil cytoplasmic tail. AB - The conformation adopted in solution by the cytoplasmic tail of CD3-epsilon has been analyzed by 1H-nmr. The cytoplasmic tail is mostly random coil expect for the amino acids conforming the immunoreceptor tyrosine-based activation motif (ITAM), YxxL/IxxxxxxxY xxL. Although the N-terminal Y xxL sequence of the motif is poorly folded, adopting 6-residue turn-like conformations with the Tyr side chain in two different orientations, the C-terminal Y xxL sequence is placed in a more complex structure involving a set of nonclassical alpha-helix turns and beta turns that comprises 11 amino acids. This structure is not modified by phosphorylation of the tyrosine residue. The differences in the conformation adopted around the two tyrosines of the ITAM motif suggest that they may play different roles pertaining to either binding signal transducing proteins or, alternatively, proteins involved in other processes such as endoplasmic reticulum location. PMID- 9209157 TI - Hydrophobic forces are responsible for the folding of a highly potent natriuretic peptide analogue at a membrane mimetic surface: an NMR study. AB - A conformational study by nmr spectroscopy was performed with the highly active 28 residue hybrid natriuretic peptide analogue pBNP1 [M. Mimeault, A. De Lean, M. Lafleur, D. Bonenfant, and A. Fournier (1995) Biochemistry, Vol. 34, pp. 955 964], which consists of the cyclic peptide core of pBNP32 and the N- and C terminal exocyclic segments of rANP (99-126). In purely aqueous solution pBNP1 exhibits random coil behavior as evidenced by the almost complete absence of structurally significant nmr observables. By contrast, elements of secondary structure emerged upon the addition of dodecylphosphocholine micelles to the aqueous sample. Nuclear Overhauser effect distance-restrained molecular dynamics simulations in conjunction with torsional angle determinations permitted the generation of reasonable model of the lipid-bound conformation of pBNP1. According to this model, pBNP1 adopts turn-like features in the cyclic and C terminal regions of the peptide, but remains quite flexible in the N-terminal segment. Two hydrophobic cores separated by a hydrophilic cleft were also evident in the generated structure. A mechanism is proposed whereby the hydrophobic interactions necessary to stabilize a folded structure of pBNP1 are facilitated by the presence of the membrane-like polar/apolar interface provided by the phospholipid micelles. PMID- 9209159 TI - Tapping mode Atomic Force Microscopy of scleroglucan networks. AB - Tapping mode Atomic Force Microscopy (TmAFM) has been used to study the fungal polysaccharide scleroglucan deposited from aqueous solution and dimethyl sulfoxide (DMSO) onto a mica surface. The solutions from which the microscope samples were produced were prepared by first dissolving the solid scleroglucan in 0.1M NaOH, then neutralizing the solution with HCl, followed by dilution to the required concentration in either water or DMSO. It was found that from the aqueous solution described above, scleroglucan forms networks. Based on a comparison of the denatured-renatured and aqueous solution samples, network formation is due to the imperfect registration between the chains forming the triple helices. The relatively large stiffness of the scleroglucan triple helix is also assumed to contribute to the formation of the extended networks. The triple helix diameter was measured to be 0.92 +/- 0.27 nm, which is in the same range as that obtained by other researchers using similar techniques. Denatured scleroglucan, deposited from DMSO onto mica, forms a web-like layer on top of which there are sphere-like structures. These morphologies are believed to be due to triple helix denaturation yielding highly flexible single chains in DMSO, which results in coiling and web-like dense packing of scleroglucan upon deposition onto mica. Most interestingly after additional of water to the samples deposited from DMSO, some of the chains can be renatured into short, stiff rod like structures which are similar to the structures observed by others researchers. The imaging data for aqueous solution deposition can be analyzed by plotting maximum end-to-end distance versus the perimeter of the networks deposited onto mica. This yields a Flory-like exponent of 0.67, which is almost similar in value to that obtained by other researchers for linear structures of scleroglucan but less than that expected for a polymer chain following a self avoiding walk (upsilon = 0.75) model on a two-dimensional surface. The fractal dimension that can be used to characterize the networks was determined graphically to be 1.22 +/- 0.06. PMID- 9209160 TI - The believability of hearsay testimony in a child sexual assault trial. AB - Two experiments investigated how mock jurors react to hearsay testimony in a case involving child sexual assault. Participants read fictional criminal trial summary involving the sexual assault of a 4-(Experiment 2 only), 6-, or 14-year old female. The summaries were presented in one of four conditions: (a) child condition--the alleged victim testified; (b) hearsay condition--the alleged victim did not testify, but an adult hearsay witness did testify; (c) multiple condition (Experiment 1 only)--both the alleged victim and the adult hearsay witness testified; and (d) no-witness condition--neither the alleged victim nor the hearsay witness testified. The hearsay testimony was believed to a considerable degree, and this testimony led to an increase in the perceived guilt of the defendant. Moreover, these results were comparable to those of conditions in which the alleged victim testified. The results are discussed in terms of the psychosocial factors affecting the perception of hearsay testimony in a child sexual assault trial. PMID- 9209161 TI - The impact of insanity acquittees on Missouri's public mental health system. AB - Even though state departments of mental health have primary responsibility for the care, custody, and treatment of insanity acquittees, the impact of insanity acquittees on the public mental health system is generally lacking in policy discussions and as a topic for policy research. This issue has received increased attention in Missouri, where insanity acquittees now occupy half of the long-term public psychiatric hospital beds. This article examines the presence of Missouri's insanity acquittees on the state's public mental health system and includes the impact on goals, fiscal costs, inpatient and community psychiatric services, and inpatient treatment staff. As states consider managed care and other cost containment measures, it remains to be seen if the high costs associated with extensive use of hospitalization of insanity acquittees to promote public safety will influence policy changes to more community-based insanity acquittee systems. PMID- 9209162 TI - Prediction of percentage body fat from anthropometric measurements: comparison of New Zealand European and Polynesian young women. AB - The prediction of total body fat from simple anthropometric measurements was examined in 42 white (New Zealand European and 40 Polynesian women aged 18-27 y. Percentage body fat (%BF) was determined from measurements of total body water (TBW) by 18O dilution. Mean (+/- SD) body mass index (BMI; in kg/m2) averaged 29.2 +/- 7.9 (range: 16.5-48.0) for the New Zealand European group and 31.2 +/- 7.9 (range: 19.8-51.8) for the Polynesian group, %BF calculated from TBW was similar in the two groups (40.5 +/- 9.9% for the New Zealand European compared with 39.1 +/- 7.5% for the Polynesian group). BMI was significantly correlated with height in the Polynesian group but not in the New Zealand European group. The relation between BMI and %BF was curvilinear for both groups. At a fixed %BF, BMI was higher in the Polynesian group than in the New Zealand European group. A BMI of 30 for the New Zealand European group corresponded to a BMI of 34 for the Polynesian group at an equivalent %BF (42%). Prediction equations for %BF developed from skinfold thicknesses or girth measurements were ethnicity dependent. We conclude that the BMI criterion for obesity in whites requires revision for use in Polynesians. PMID- 9209163 TI - The fatness-specific bioelectrical impedance analysis equations of Segal et al: are they generalizable and practical? AB - The fatness-specific bioelectrical impedance analysis (BIA) equations of Segal et al (Am J Clin Ntr 1988;47: 7-14; Segal equations) have been shown to be generalizable across sex, ethnicity, age, and degrees of adiposity. However, these fatness-specific equations require an a priori determination of percentage body fat (%BF) by using a skinfold equation or densitometry to categorize subjects into obese or nonobese groups. These procedures negate the use of BIA as a fast and simple method. It was hypothesized that the average of the Segal nonobese and obese fatness-specific equations (BIA average method) could be used in lieu of the skinfold method for categorizing subjects who are not obviously lean or obese. In phase 1 these three methods were compared for a subsample of 59 women who were not obviously lean or obese. The %BF of 75% of these subjects was accurately estimated within 3.5%BF by using the BIA average method whereas only 71% and 46% were accurately estimated by fusing the densitometric and skinfold methods, respectively. In phase 2, the predictive accuracy of the Segal fatness specific equations, used in combination with the BIA average method, was compared with other BIA equations published previously for 602 American Indian, Hispanic, and white women and men. The Segal fatness-specific equations yielded the smallest prediction error (SEE = 2.22 kg for women and 3.59 kg for men) and the %BF of 70% of the subjects was accurately estimated within 3.5%BF compared with 24-59% for other BIA equations. Therefore, we recommend using the Segal fatness specific and average equations to assess body composition in heterogeneous populations. PMID- 9209164 TI - Between-day and within-day variability in the relation between heart rate and oxygen consumption: effect on the estimation of energy expenditure by heart-rate monitoring. AB - Estimation of energy expenditure (EE) by heart-rate (HR) monitoring (HRM) assumes that the relation between HR and oxygen consumption (VO2) is stable between days and within a day. To evaluate this assumption, 12 healthy subjects underwent an HR-VO2 calibration session on two mornings and two afternoons, with one morning and one afternoon session on the same day. Measurements were made while subjects were supine, sitting, standing while shifting body weight side-to-side, and walking at four intensities. Subjects wore an HR monitor during waking hours on another day (15.1 +/- 1.5 h). Regression analysis was used to determine the relation between HR and VO2 in the sedentary and active HR ranges, and four EE values (HRM-EE) based on the four calibration sessions were calculated for each subject. The four group mean HRM-EE values were nearly identical (CV: 1.1%). The regression equations generated from the four calibration sessions did not differ significantly for the group as a whole, but for some subjects there were significant differences among sessions in the slope of the active regression equation (P = 0.005). Intraindividual CVs for HRM-EE were generally < 10%, but ranged from 0.1% to 24.7%. In general, within an individual, HR was more variable than was VO2, and intraindividual variability in EE was associated with intraindividual variability in the flex HR and sedentary HR range. HRM is appropriate for assessment of EE for a group; however, caution is recommended when HRM is used for individual determinations of EE. PMID- 9209165 TI - Reduced food intake after jejunoileal bypass: a possible association with prolonged gastric emptying and altered gut hormone patterns. AB - The object of this study was to examine whether eating behavior, food preference, gastric emptying, and gut hormone patterns are altered after jejunoileal bypass (JIB) in patients with severe obesity. Eight obese [mean (+/- SD) body mass index (BMI; in kg/m2) 42.9 +/- 4] subjects were studied prospectively before and 9 mo after JIB with eight age- and sex-matched normal-weight control subjects. Total energy intake, data from the universal eating monitor (VIKTOR), eating motivation measured by visual analog scales, a food-preference checklist, a forced-choice list, solid-phase gastric emptying, and postprandial concentrations of cholecystokinin, motilin, and neurotensin were studied. BMI was reduced by 29% after JIB. Compared with normal subjects, the JIB patients showed a reduced desire to eat, decreased hunger, and reduced prospective consumption before a test meal. After surgery, obese subjects selected fewer food items and showed a reduced preference for high-carbohydrate and high-fat items before a test meal. There was a trend from an accelerated toward a decelerated eating pattern in obese subjects after JIB. After JIB, gastric emptying of obese subjects was slowed and similar to that in control subjects. Obese subjects had lower postprandial cholecystokinin concentrations that were lower than those of control subjects both before and after JIB. Postprandial concentrations of neurotensin were higher after JIB. We conclude that after JIB, the desire to eat and preference for high-carbohydrate and high-fat items is reduced, resulting in decreased energy intake. That gastric emptying is prolonged and gut hormone patterns are altered with low postprandial plasma cholecystokinin and high neurotensin plasma concentrations may at least partly account for these observations. PMID- 9209166 TI - Response of serum leptin concentrations to 7 d of energy restriction in centrally obese African Americans with impaired or diabetic glucose tolerance. AB - The aim of this study was to determine whether serum leptin concentrations are reduced in response to short-term energy restriction in centrally obese individuals with impaired glucose tolerance or non-insulin-dependent diabetes mellitus. Twenty African Americans [16 females and 4 males, 44 +/- 7 y (x +/- SD), 107.2 +/- 23.8 kg, 39 +/- 7% body fat] consumed a 7-d energy-restricted diet (4.03 +/- 0.72 MJ/d) of whole foods. Oral-glucose-tolerance tests (OGTTs) were performed before and immediately after the diet to assess changes in serum leptin, glucose, and insulin concentrations. Baseline leptin concentration correlated significantly with percentage body fat (r = 0.80), body mass index (r = 0.72), fat mass (4 = 0.64), waist-height ratio (r = 0.6), body weight (r = 0.59, all P < 0.01), waist circumference (r = 0.49), and basal insulin concentration (r = 0.48, both P < 0.05). Seven days of energy restriction resulted in significant reductions (P < 0.005) in leptin (-6.1 +/- 8.4 micrograms/L), basal glucose (-0.9 +/- 0.8 mmol/L), OGTT glucose area under the curve (-158 +/- 164 mmol/L), and basal insulin concentration (-34 +/- 69 pmol/L, P < 0.05). In addition, there was a trend for a reduction in OGTT insulin area under the curve (-15,567 +/- 3,658 pmol/L, P = 0.05), and a tendency for basal insulin and leptin to change together (r = 0.41, P = 0.07). Despite the weight loss of 3.1 +/- 1.3 kg (P < 0.0001), the loss of fat mass was calculated to be only -1.0 +/- 0.1 kg. These results suggest that negative energy balance or improved insulin action was responsible for the changes in leptin, glucose, and insulin concentrations. In summary, short-term energy restriction effectively reduced serum leptin concentrations and improved glucose tolerance and insulin action in obese individuals with impaired or diabetic glucose tolerance. PMID- 9209167 TI - Correlates of hypoalbuminemia in community-dwelling older persons. AB - To identify easily ascertainable sociodemographic and health characteristics that are associated with hypoalbuminemia in community-dwelling older persons, we used data from the first National Health and Nutrition Examination Survey. This population-based stratified probability sample survey included 4728 persons aged 55-74 y. We defined hypoalbuminemia in two ways: < 35 g/L (1.2% of the sample) or < or = 38 g/L (7.9% of the sample) and used multivariate logistic models to identify independent predictors of hypoalbuminemia. Older age; receiving welfare; a condition interfering with eating; vomiting > or = 3 d/mo; previous surgery for gastrointestinal tumor; self-reported heart failure; recurring cough attacks; feeling tired or wornout; edentulous, fair, or poor condition of teeth; little or no exercise; a low-salt diet; trouble chewing meat; self-reported protein albumin, blood, or sugar in urine; and current cigarette smoking were independently associated with albuminemia (< or = 38 g/L) or progressively lower albumin concentrations < 40 g/L. Persons with 3-5 of these factors (51.5% of the sample) had an odds ratio of 2.73 (95% CI: 1.64, 4.54) and those with > or = 6 factors (9.4% of the sample) had an odds ratio of 6.44 (95% CI: 3.49, 11.86) of albuminemia < or = 38 g/L compared with those with 0-2 risk factors (39.1% of the sample). These findings suggest that several easily assessed sociodemographic, lifestyle, and disease-related factors are associated with hypoalbuminemia and might be valuable items to include on general health surveys to identify older persons who have this marker of poor health status. PMID- 9209168 TI - Urinary isoflavonoid excretion in humans is dose dependent at low to moderate levels of soy-protein consumption. AB - Soybeans contain isoflavones, which have been associated with many health benefits, including decreased cancer risk. The purpose of our study was to measure urinary isoflavonoid excretion in response to daily consumption of soy that contained 0-36 mg isoflavones--a lower range than used in previous studies- and to compare urinary isoflavonoid excretion between equol excreters and nonexcreters. Fourteen men and women aged 20-40 y participated in the study. Half of the subjects were identified previously as equol excreters and the other half as equol nonexcreters. This randomized, double-blind, crossover study consisted of four 9-d diet treatment periods. During each treatment period participants consumed a low-photoestrogen controlled diet and a beverage containing 0, 5, 10, or 20 g soy protein. Urine collected on the last 3 d of each treatment period was analyzed for isoflavonoid (equol, O-desmethylangolensin, genistein, and daidzein) and lignan (enterodiol and enterolactone) contents by using isotope-dilution gas chromatography-mass spectrometry. There was a highly linear dose response of urinary isoflavonoid excretion to soy consumption, which did not differ significantly between equol excreters and nonexcreters. There were no significant differences in lignan excretion between the two diet treatments. Our results indicate that urinary isoflavonoid excretion is dose dependent in humans at low to moderate levels of soy consumption. PMID- 9209169 TI - beta-Carotene in breast milk and serum is increased after a single beta-carotene dose. AB - Normal lactating mothers were administered a single dose of 60 or 210 mg beta carotene and changes in serum and milk retinol, alpha-tocopherol, and carotenoids were monitored for 8 d. Average serum beta-carotene concentrations increased 4.1- and 4.0-fold after the 60- and 210-mg doses, respectively. Milk beta-carotene concentrations increased 4.1- and 3.0-fold after the 60- and 210-mg doses, respectively. Maximum serum concentrations were reached 24 h after both supplements, although concentrations of milk beta-carotene continued to rise for 2-3 d. After 8 d, both serum and milk beta-carotene continued to rise for 2-3 d. After 8 d, both serum and milk beta-carotene concentrations remained about twofold higher than baseline concentrations. Increases in serum or milk beta carotene concentrations were not dose-dependent. Initial serum and milk concentrations of beta-carotene predicted increases after supplementation, and increases in serum beta-carotene concentrations predicted those in milk. Concentrations of milk carotenoids were less than one-tenth their respective concentrations in serum. Lutein, beta-cryptoxanthin, lycopene, alpha-carotene, retinol, and alpha-tocopherol concentrations in serum or milk did not change significantly after beta-carotene supplementation. Retinol esters account for most of the retinol equivalents in the milk of well-nourished mothers. Initial and maximum concentrations of beta-carotene in serum and milk were strongly correlated for individual mothers. Collectively, the data showed that a single 60 mg supplement of beta-carotene sustained elevated beta-carotene concentrations in serum and milk for > 1 wk in normal mothers but did not affect concentrations of other major carotenoids, retinol, or alpha-tocopherol. PMID- 9209170 TI - Maldigestion and colonic fermentation of wheat bread in humans and the influence of dietary fat. AB - A fraction of wheat bread is malabsorbed in healthy humans. The malabsorbed fraction is bigger than what can be accounted for by in vitro measurements of dietary fibers and resistant starch. To determine whether it is a specific fraction defined by the structure of the starch molecule or a variable amount- which depends on the individual, the amount ingested, and other components of the meal--we performed a dose-response study on wheat bread in healthy human volunteers. Malabsorption was evaluated by using the breath-hydrogen test. Test meals were as follows: 20 g wheat bran mixed in 100 mL water; bread made from 25, 75, 100, 150, and 200 g white wheat flour (WWF); bread made from 0 g WWF and 20 g wheat bran; and bread made from 100 g WWF served with 11 or 26 g butter, corresponding to 20% or 35% of energy from fat in the meals. Three of seven volunteers malabsorbed a fraction of the bread made from 25 g WWF and five of seven a fraction of the bread made from 75 g WWF. All volunteers malabsorbed a fraction of the 100-g WWF bread, Bread made from 180 g WWF and 20 g wheat bran resulted in a breath-hydrogen response of the same magnitude as that from bread made from 200 g WWF alone. The 100-g WWF bread + 11 g butter resulted in a significantly higher breath-hydrogen response than did the bread alone, whereas the 100-g WWF bread + 26 g butter resulted in an average response of the same magnitude as that from bread alone. We conclude that the malabsorbed fraction of wheat bread was dependent on the amount ingested, the composition of the meal, and individual gastrointestinal handling. Fermentation of wheat bran resulted in a very low breath-hydrogen response compared with lactulose or wheat bread. Addition of 11 g butter to the bread seemed to increase the malabsorbed fraction of the starch, an effect that was abolished when the amount of butter was increased to 26 g. PMID- 9209171 TI - Assessment of vitamin A status by the deuterated-retinol-dilution technique and comparison with hepatic vitamin A concentration in Bangladeshi surgical patients. AB - Hepatic stores of vitamin A were estimated in 31 Bangladeshi surgical patients (15 males and 16 females) by the deuterated-retinol-dilution (DRD) technique and by analysis of the vitamin A concentration of a liver biopsy specimen obtained during previously scheduled abdominal surgery. Patients ranged in age from 21 to 65 y and had an average body mass index (BMI: in kg/m2) of 17.7 +/- 3.4. They received 0.753 mumol [2H4]retinyl acetate/kg body wt orally 9-11 d before surgery. Hepatic vitamin A reserves were estimated according to Furr et al (Am J Clin Nutr 1989;49:713-6) by using a single plasma isotopic-ratio measurement (18 25 d postdose). Estimated mean hepatic vitamin A stores were similar by both techniques, 0.110 +/- 0.072 mmol (by DRD) compared with 0.100 +/- 0.067 mmol (by biopsy). Regression analysis was used to compare results of the DRD and biopsy techniques. A significant linear relation was found between the two techniques (r = 0.75, P < 0.0001), and the least-squares regression line was not significantly different from y = x (P = 0.09). The results indicate that the DRD technique provided a very good estimate of hepatic vitamin A reserves for this population. However, a wide prediction interval was observed for estimates of hepatic vitamin A reserves for individual subjects. Thus, further refinement of the prediction model is necessary to improve estimates of hepatic vitamin A reserves for individual subjects. PMID- 9209172 TI - Acute functional iron deficiency in obese subjects during a very-low-energy all protein diet. AB - We examined whether a very-low-energy all-protein diet (VLED) would produce detectable changes in iron as well as in other trace elements. Twenty-five obese patients consumed for 2 wk a VLED containing 70 g protein after a 1-wk period during which total daily energy intake was progressively reduced to 1.26 MJ. Serum iron fell sharply by approximately equal to 50% (P < 0.0001), and despite a small decrease in total-iron-binding capacity, transferrin saturation decreased from 30 +/- 11% to 18 +/- 5% (P < 0.0001). Serum ferritin did not change significantly but serum soluble transferrin receptor (sTfR), an indicator of iron deficiency, increased progressively from 4630 +/- 1110 to 6070 +/- 1390 micrograms/L (P < 0.0001). Changes in sTfR correlated inversely with prior changes in serum iron. Changes in iron metabolism did not translate into changes in erythropoiesis or red cell indexes, but the white blood cell count decreased from 7.3 +/- 1.6 to 6.2 +/- 1.9 x 10(9)/L (P < 0.002). There was no evidence of deficiency for the other trace elements and minerals tested. Daily supplementation with 200 mg Fe in 18 other subjects only partially corrected these observations despite some increase in iron stores. These results indicate that during a 2-wk VLED serum iron is significantly depressed, inducing functional tissue iron deficiency too short in duration to produce alterations in red blood cell indexes. These changes are not mediated by absolute iron deficiency, inflammation, or protein malnutrition but could be related to alterations in the iron storage and release behavior of the reticuloendothelial cell during energy deprivation alone. PMID- 9209173 TI - Zinc absorption in women during pregnancy and lactation: a longitudinal study. AB - Zinc is essential for normal fetal growth and development and for milk production during lactation. The metabolic adjustments made in zinc utilization to meet these needs have not been described. The purpose of this study was to determine whether fractional zinc absorption (FZA) is altered during pregnancy and lactation and, if so, to determine whether the change is related to maternal zinc status, specifically, concentrations of zinc in plasma, erythrocytes, urine, and breast milk and dietary zinc intake. Thirteen women were studied at five time points: once preconception; at 8-10, 24-26, and 34-36 wk gestation; and once while they were lactating 7-9 wk postpartum. Zinc intake increased by 35 mumol/d (2.3 mg/d) from preconception to 34-36 wk (P = 0.04); it tended to decrease (P > 0.05) during lactation but did not return to the preconception level. The amount of zinc in breast milk averaged 2.0 mg/d at the lactation time point. FZA measured from urinary enrichments of two stable isotopes of zinc increased from 14% preconception to 25% during lactation (P = 0.023) but the increase to 19% at 34-36 wk gestation was not significant. No increase in FZA occurred in four women who took iron supplements during lactation. FZA was negatively correlated with plasma zinc concentration at 34-36 wk gestation and with urinary zinc excretion at all time points. The nearly twofold increase in zinc absorption during lactation was presumably in response to the demand for zinc to synthesize breast milk. PMID- 9209174 TI - Does vegetable oil attenuate the beneficial effects of fish oil in reducing risk factors for cardiovascular disease? AB - Contradictory reports on the protective effect of fish consumption on cardiovascular disease (CVD) risk could be due to variations in the intake of n-3 and n-6 polyunsaturated fatty acids (PUFAs). Metabolic competition between n-3 and n-6 PUFAs suggests that n-6 PUFAs in vegetable oils could attenuate the efficacy of n-3 PUFAs in fish oil to favorably alter endpoints relevant to CVD risk. We determined the effects of varying dietary amounts of fish oil on lipid and thrombotic endpoints relevant to risk factors for CVD and whether these effects were attenuated by vegetable oils. Two randomized, double-blind, placebo controlled, parallel studies were conducted in human subjects fed varying amounts of n-3 and n-6 PUFAs; n-3 PUFA intake was varied by using fish or placebo oil capsules, and n-6 PUFA intake was modified by incorporating varying amounts of safflower oil into the diet. Endpoints included changes in membrane fatty acid composition, blood lipids, and thrombotic profile. The results indicated that absolute amounts of fish oil, and not the relative amounts of fish and vegetable oil (ratios of n-3 to n-6 PUFAs), determined the magnitude of the reduction of arachidonic acid and increase in eicosapentaenoic acid in phospholipids of plasma and platelets. The suppression of plasma triacylglycerols by fish oil was not affected by varying amounts of dietary n-6 PUFAs. Fibrinogen concentrations decreased with 15 g but not with 9 g fish oil/d fed at the same ratio of n-3 to n 6 PUFAs. The efficacy of fish oil in favorably modifying certain risk factors for CVD was not attenuated by vegetable oil. PMID- 9209175 TI - Influence of the SstI polymorphism at the apolipoprotein C-III gene locus on the plasma low-density-lipoprotein-cholesterol response to dietary monounsaturated fat. AB - The plasma lipid response to changes in dietary fat and cholesterol can vary between individuals. The SstI polymorphism, arising from a cytosine to guanosine substitution in the 3' untranslated region of the APOC3 gene distinguishes between two alleles--S1 and S2. The S2 allele has been associated with elevated plasma triacylglycerol, cholesterol, and apolipoprotein (apo) C-III concentrations. In 90 young men we examined the effect of the same mutation on the response of low-density-lipoprotein (LDL) cholesterol to dietary monounsaturated fat. The frequency for the S2 allele was 0.14. Subjects were fed a low-fat diet for 25 d, followed by a diet rich in monounsaturated fatty acid (22% MUFA, 38% total fat) for 28 d; lipoproteins were measured at the end of each diet. There were no significant differences in initial total cholesterol between subjects with the APOC3*S1/APOC3*S1 (S1/S1) and APOC3*S1/APOC3*S2 (S1/S2) genotypes. After consumption of the diet high in MUFA, significant increases in LDL cholesterol (0.13 mmol/L, P < 0.027) were noted in the S1/S1 subjects whereas a significant decrease was observed in the S1/S2 subjects (-0.18 mmol/L, P < 0.046). Significant genotypic effects were seen for diet-induced changes in LDL cholesterol (P < 0.00034), total cholesterol (P < 0.009), and apo B (P < 0.0014). A study of the effect of the interaction between this mutation with that present in position -76 of the APOA1 gene promoter region (G/A) revealed that both mutations had an additive effect on changes in total cholesterol, LDL cholesterol, and apo B induced by diets. Plasma LDL-cholesterol responsiveness to the diet may be explained, at least in part, by variation at the APOC3 gene locus. PMID- 9209176 TI - Changes in iron status during weight loss with very-low-energy diets. AB - For several decades, very-low-energy diets (VLEDs) have been used by obese individuals to achieve weight loss. During the weight loss, patients often have dramatic drops in circulating thyroid hormone concentrations and experience cold intolerance. Because poor iron status is known to alter thermogenesis, we investigated the possibility that iron intake interacts with energy intake during weight loss in obese individuals. The effects on indicators of iron and thyroid status of increasing the iron content of a VLED from 18 to 27 mg/d during 12 wk of a VLED were compared with the effects on the same indicators of increasing energy intake from 1752 kJ(420 kcal) to 3347 kJ(800 kcal)/d. Although all VLED groups initially had 30% declines in plasma transferrin saturation, increases in plasma ferritin concentrations, and decreases in plasma thyroid hormone concentrations, patients who received iron supplementation had significantly higher circulating concentrations of triiodothyronine and thyroxine at the end of the VLED than did patients who received only the recommended dietary allowance of iron. The patients who received iron supplementation also had a more rapid return of iron indicators to normal values over the course of the VLED. The transitory fall in iron delivery to bone marrow was not associated with anemia. These data suggest that higher thyroid hormone concentrations can be maintained during VLEDs that provide higher iron intakes. PMID- 9209178 TI - Lycopene is more bioavailable from tomato paste than from fresh tomatoes. AB - Lycopene bioavailability from a single dose of fresh tomatoes or tomato paste (23 mg lycopene) ingested together with 15 g corn oil was compared by analyzing carotenoid concentrations in the chylomicron fraction. The lycopene isomer pattern was the same in both fresh tomatoes and tomato paste. The triacylglycerol response in chylomicrons was not significantly different after both treatments. Ingestion of tomato paste was found to yield 2.5-fold higher total and all-trans lycopene peak concentrations (P < 0.05 and P < 0.005, respectively) and 3.8-fold higher area under the curve (AUC) responses (P < 0.001) than ingestion of fresh tomatoes. The same was calculated for lycopene cis-isomers, but only the AUC response for the cis-isomers was significantly higher after ingestion of tomato paste (P < 0.005). No difference was observed in the alpha- and beta-carotene response. Thus, in humans, the bioavailability of lycopene is greater from tomato paste than from fresh tomatoes. PMID- 9209177 TI - Effects of age on body fat distribution and cardiovascular risk factors in women. AB - We conducted a cross-sectional study of body fat distribution and metabolic variables and the interrelations among these factors in 134 women aged 18-71.9 y. Body fat distribution was measured with use of computerized tomography. A significant positive correlation was observed between age and visceral adipose tissue (VAT) and between VAT and body weight. When subjects were divided into five age groups, VAT values were significantly higher in older groups. Values for triacylglycerols, cholesterol, fasting glucose, 2-h glucose, and the sum of glucose values during an oral-glucose-tolerance test were significantly higher in older subjects. After adjustment for visceral fat, no significant differences in any metabolic variable studied, except cholesterol, were found across the five age groups. In conclusion, we found that regional body fat distribution in older women was different from that in younger subjects: older women had larger amounts o visceral fat. Values for metabolic variables were also higher in older subjects. Our data suggest that redistribution of body fat in older subjects is associated with changes in metabolic variables. PMID- 9209179 TI - Effects of milk viscosity on gastric emptying and lactose intolerance in lactose maldigesters. AB - The possibility of delaying gastric emptying and improving lactose digestion and tolerance by increasing milk viscosity was studied in 13 lactose maldigesters who ingested three test milks with different viscosities (range: 33-1892 mPa.s) in random order at intervals of 1 wk. Each test portion was 500 mL and provided approximately equal to 1900 kJ and 18 g lactose. The different viscosities were obtained by adding varying proportions of rice starch and maltodextrin to a basic milk formula. A combined [13C]glycine-hydrogen breath test was used to measure gastric emptying and lactose digestion simultaneously. Participants reported their gastrointestinal symptoms by using a four-grade scale. Mean (+/- SEM) gastric-emptying half times were 78 +/- 5.7 min for low-viscosity milk (30 mPa.s), 86 +/- 5.0 min for moderate-viscosity milk (80 mPa.s), and 78 +/- 4.5 min for high-viscosity milk (1.9.10(3) mPa.s). Mean orocecal transit times (180 +/- 24, 163 +/- 23, and 180 +/- 24 min, respectively) were not significantly different. There were no milk-dependent differences in breath-hydrogen excretion or in the severity of gastrointestinal symptoms. The milks were well tolerated; > 50% of the subjects reported nondisturbing symptoms or none. We conclude that gastric emptying, orocecal transit time, and lactose digestion and tolerance were not affected by altering milk viscosity. This may have been due to the high energy content of the test milks, which in itself led to slow gastric emptying. PMID- 9209180 TI - Relation between postprandial satiation and antral area in normal subjects. AB - The factors influencing appetite in humans are poorly understood. There is a weak relation between appetite and gastric emptying in normal subjects. Recent studies have shown that fasting and postprandial antral areas increase in patients with functional dyspepsia compared with normal subjects. We evaluated the hypothesis that antral area, and hence antral distention, is a significant determinant of postprandial fullness. Fourteen normal subjects had simultaneous measurements of gastric emptying by scintigraphy and antral area by ultrasound after ingestion of 350 mL 20% glucose. Fullness and hunger were assessed by visual analog scales. Measurements of the gastric-emptying half time (t1/2) by scintigraphy and ultrasound were not significantly different (129.6 +/- 11.8 min compared with 115.6 +/- 11.4 min). Fullness increased (P < 0.001) and hunger decreased (P < 0.001) after the drink. Both fullness and the magnitude of the increase in fullness after the drink were related to antral area (r > 0.56, P < 0.05), the increase in antral area (r > 0.59, P < 0.05), and the scintigraphic content of the distal stomach (r > 0.57, P < 0.05), but not to the ultrasound or scintigraphic t1/2 values. In contrast, hunger and the magnitude of the decrease in hunger after the drink were not related to either antral area, the increase in antral area, or the rate of gastric emptying. We conclude that postprandial fullness, but not hunger, was closely related to antral distention in normal subjects. PMID- 9209181 TI - Increase in diet-induced thermogenesis at the start of refeeding in severely malnourished anorexia nervosa patients. AB - Many reports describe the difficulty for anorexia nervosa patients to gain weight during refeeding. To assess whether an increase in diet-induced thermogenesis (DIT) participates to this resistance, we studied DIT by indirect calorimetry in 11 severely malnourished anorexia nervosa patients [body mass index (BMI; in kg/m2) = 13] to accomplish two purposes: 1) to compare DIT in a strict semistarvation state with that obtained after 1 wk refeeding, when metabolism is shifted to a dynamic trend toward regaining weight, without significant change in body composition; 2) to study the effect on DIT of two energetic loads representing each one-third of the energy intake during semistarvation and refeeding, respectively: 1.25 and 2.92 MJ. To avoid bias, the two liquid loads were infused intragastrically in a random double-blind fashion. A significant increase in DIT during refeeding was observed for the two loads (204 +/- 23 kJ for the 1.25-MJ liquid meal and 482 +/- 78 kJ for the 2.92-MJ one, P < 0.02). The higher the load, the larger the increase with refeeding (P < 0.001). This increment in DIT exceeded the increase in active lean body mass and was poorly correlated with lean body mass. These results provide clear evidence of a strong cellular "waste" mechanism in anorexia nervosa patients during the early phase of refeeding, which enhances the adaptative resistance to overfeeding that we have already shown for resting energy expenditure. PMID- 9209182 TI - Computerized tomography assessment of women with weight changes associated with adjuvant treatment for breast cancer. AB - It is common for women undergoing treatment for breast cancer to gain weight, although the characteristics of the weight change have not been described. We investigated the changes in abdominal fat accumulation that accompanied the change in weight associated with treatment for breast cancer in longitudinal and cross-sectional clinical studies in 34 women aged 39-73 y with early-stage primary breast cancer. Computerized tomography scans of abdominal subcutaneous and visceral adipose depots, bioelectrical impedance measurements of body fat mass, and measurements of body weight and girth were obtained early in the course of treatment and 6 mo later (longitudinal study; n = 8) or within 12 mo of treatment (cross-sectional study; n = 26). The longitudinal study found that, irrespective of the direction of weight change, seven of eight women gained body fat and lost lean body mass. In the five women who gained weight (median: 3.2 kg) two lost and three gained subcutaneous adipose fat (median: 19%) whereas all gained visceral fat (median: 23%). In the cross-sectional study 19 women gained weight and 7 lost weight or had stable weight since diagnosis. Change in weight was correlated with abdominal subcutaneous adipose fat (r = 0.39; P = 0.06) and hip circumference (r = 0.43; P = 0.03) but not abdominal visceral fat, the ratio of subcutaneous to visceral fat, or the ratio of waist to hip size. In the longitudinal sample, weight gain resulted in a variable response in subcutaneous adipose volumes but a consistent increase in visceral adipose depot. Although these results are preliminary, it appeared that regardless of weight gain or loss women were likely to lose lean body mass and gain fat mass during treatment for breast cancer. PMID- 9209183 TI - Alterations in lymphocyte subsets and pituitary-adrenal gland-related hormones during fasting. AB - We investigated changes in the immunoendocrine system during fasting. Ten hospitalized patients aged 14-46 y with psychosomatic disorders fasted for 7 or 10 d. Blood samples were collected before and on days 3 and 7 of the 7-d fasts. When fasting continued to 10 d, an additional sample was taken on day 10. We measured blood cellularity (white blood cells and total lymphocytes), the total number and percentage of lymphocyte subsets (CD2, CD3, CD4, CD8, and CD19), natural killer (NK) cell activity, cytokines (interleukin 1 beta, interleukin 2, interleukin 6, granulocyte-macrophage colony stimulating factor, tumor necrosis factor alpha, and interferon gamma), and soluble interleukin 2 receptors. Corticotropin, cortisol, and dehydroepiandrosterone sulfate (DHEAS) concentrations were also determined. Although the total number of lymphocytes decreased during fasting, NK cell activity increased significantly. Plasma cortisol and DHEAS concentrations also increased significantly whereas changes in corticotropin concentrations were not significant. The total number and percentage of CD4 cells decreased significantly during fasting but no other lymphocyte subsets changed significantly. The percentage of CD4 cells was negatively correlated with cortisol concentrations during fasting. No detectable changes occurred in cytokines or soluble interleukin 2 receptors during the study. All measured immunoendocrine values that changed during fasting returned to prefasting values during the refeeding period. These findings indicate that fasting affects immune variables such as T cell subsets and NK cell activity at least in part through changes in adrenal gland-related hormones. PMID- 9209184 TI - Carbohydrate supplementation affects blood granulocyte and monocyte trafficking but not function after 2.5 h or running. AB - This randomized, double-blind, placebo-controlled study was designed to determine the influence of carbohydrate supplementation on the granulocyte and monocyte response to 2.5 h of high-intensity running [76.7 +/- 0.4% of maximal oxygen consumption (VO2max)]. Thirty experienced marathon runners (VO2max 53.4 +/- 1.0 mL.kg-1.min-1, age 41.5 +/- 1.4 y) were randomly assigned to carbohydrate supplement (n = 17) and placebo (n = 13) groups. Subjects rested for 10-15 min before a blood sample was taken at 0715, and then ingested 0.75 L carbohydrate beverage or placebo. At 0730 subjects began running at 75-80% of VO2max for 2.5 h, and drank 0.25 L carbohydrate or placebo fluid every 15 min. Immediately after the 2.5-h run (1000), another blood sample was taken, followed by 1.5-h, 3-h, and 6-h recovery samples. Carbohydrate supplementation had a significant effect compared with placebo on the pattern of change in plasma glucose and cortisol, and the blood concentration of neutrophils (F[14, 112] = 5.13, P = 0.001) and monocytes (F[14, 112] = 4.78, P = 0.001), but not on blood granulocyte and monocyte phagocytosis or oxidative burst activity after 2.5 h of intensive running. PMID- 9209185 TI - Evaluation of biochemical indicators of vitamin A status in breast-feeding and non-breast-feeding Indonesian women. AB - Indicators of vitamin A status were evaluated in nonpregnant breast-feeding (n = 265) and nonpregnant non-breast-feeding (n = 49) Indonesian women. The concentration of vitamin A (not including provitamin A carotenoids) and fat in breast milk was 30% and 20% higher, respectively, for women with a breast-fed child 7-18 mo old than for women with an infant 3-6 mo old. The vitamin A content of milk fat was constant throughout lactation. Breast-milk vitamin A was most sensitive to changes in vitamin A status when expressed per volume. Sensitivity and specificity for detecting serum retinol concentrations < 0.70 mumol/L were < 75% for the concentration of breast-milk vitamin A and serum retinol binding protein (RBP). The modified-relative-dose-response (MRDR) method suffers from a relatively large intraindividual variation in the ratio of dehydroretinol to retinol because of vulnerability of the dehydroretinol concentration to laboratory errors and to variation in dosing and absorption. Within categories of dehydroretinol:retinol, serum retinol concentration was lower in breast-feeding women than in non-breast-feeding women. Thus, it may be necessary to use different cutoff values for the ratio and for serum retinol concentration. Serum retinol concentration, which was just above marginal (0.85 mumol/L), had the smallest within-person variation and was also the most sensitive indicator for detecting a difference between groups in change in vitamin A status postintervention, requiring only 19 subjects per group. Serum RBP concentration, breast-milk vitamin A expressed per volume or per gram milk fat, and the MRDR method required groups of 35, 36, 139, and 53 subjects, respectively. PMID- 9209186 TI - Effects of discontinuing coffee intake on iron status of iron-deficient Guatemalan toddlers: a randomized intervention study. AB - Coffee is one of the first liquids given to infants in Guatemala. To evaluate whether this practice has an adverse effect on iron status, 160 children 12-24 mo of age who had received coffee for > or = 2 mo and had at least one indicator of iron deficiency were stratified by initial hemoglobin concentration (anemic, or nonanemic, ie, hemoglobin > or = 105 g/L) and randomly assigned to a control (continuation of coffee; coffee) or intervention (provided with a substitute consisting of sugar and coloring; substitute) group for 5 mo. Anemic children were provided with iron supplements for 2-3 mo. Hematologic and anthropometric measurements were made before and after the intervention and dietary and morbidity data were collected every 2 wk. A total of 139 children completed the study: 45 coffee, nonanemic; 56 substitute, nonanemic; 19 coffee, anemic; and 19 substitute, anemic. Compliance with the procedures was good: median coffee intake was 891 mL/wk in the coffee group compared with 18 mL/wk in the substitute group (P = 0.0001). There was no significant effect of discontinuing coffee consumption on changes in hemoglobin, hematocrit, ratio of zinc protoporphyrin to heme or plasma iron, zinc or copper in either nonanemic or anemic children, or plasma ferritin in children who did not take iron supplements. In children who took iron supplements, change in plasma ferritin was significantly greater in the substitute group than in the coffee group (106% compared with 1%, P < 0.05). This implies that coffee interferes with the utilization of supplemental iron. It is likely that the amount and strength of coffee consumed by Guatemalan toddlers are too low to significantly affect the other indexes of iron status. PMID- 9209187 TI - Weekly micronutrient supplementation to build iron stores in female Indonesian adolescents. AB - Different supplementation schemes to build iron stores in female Indonesian adolescents were investigated. Subjects were 273 high-school girls allocated randomly to four treatment groups. During a 3-mo period one group received 60 mg Fe, 750 micrograms retinol, 250 micrograms folic acid, and 60 mg vitamin C per day; a second group received 60 mg Fe, 6000 micrograms retinol, 500 mg folic acid, and 60 mg vitamin C once a week; a third group received 120 mg Fe and the same amount of the other three micronutrients as the second group once a week; and a fourth group received only placebos. All subjects were dewormed and supplement allocation was double blind. Blood samples were collected at baseline, after 2 and 3 mo of supplementation, and 6 mo after the last supplement. After 2 mo of supplementation, groups supplemented weekly and daily showed similar significant improvements (P < 0.001) in hemoglobin and retinol concentrations, and supplementation for 3 instead of 2 mo did not significantly increase these two indicators. After 3 mo, the increase in ferritin was approximately equal to 27 micrograms/L in the daily and 14-15 micrograms/L in the weekly groups (P < 0.001), the latter having a final concentration of 42-45 micrograms/L. At 6 mo postsupplementation there were no significant differences among daily and weekly groups, but the ferritin concentration was still approximately equal to 10-12 micrograms/L higher (P < 0.001) than in the placebo group. The group supplemented weekly with 60 mg Fe complained less about side effects than the other supplemented groups (P < 0.05). Weekly supplementation with 60 mg Fe and 6000 micrograms retinol for 3 mo was optimal for improving the iron status of the adolescents for approximately equal to 9 mo. PMID- 9209188 TI - Obesity surgery as a model for understanding the regulation of food intake and body weight. PMID- 9209189 TI - Isotope-dilution techniques: a wave of the future in human nutrition. PMID- 9209190 TI - Do the n-3 fatty acids from fish prevent deaths from cardiovascular disease? PMID- 9209191 TI - What do indicators indicate? PMID- 9209192 TI - Standardization of nomenclature of body composition in weight loss. PMID- 9209194 TI - What should an accountable certifying board do about training programs that are substandard? PMID- 9209193 TI - Community, public, and global nutrition. PMID- 9209195 TI - The influence of parasympatholytics on the resolution of acute attacks of asthma. AB - PURPOSE: To evaluate the role of parasympatholytics in the resolution of acute attacks of asthma. METHODS: This study employed a prospective sequential design in which the influence of 0.5 and 1.0 mg of ipratropium bromide on peak expiratory flow rates (PEFR), hospital admissions, and length of stay (LOS) in the emergency department (ED) was evaluated. The parasympatholytic was added to a well-investigated standard therapeutic regimen that was anchored by the use of repetitive doses of albuterol, and employed pretested decision algorithms. RESULTS: One hundred and thirty-one patients received ipratropium (l) and 123 who did not (NI) served as controls. There were no significant pretreatment between group differences in gender, racial composition clinical signs and symptoms, or PEFR. The presence of ipratropium in the regimen did not influence discharge/admission patterns, LOS, the rate of improvement of the patients, or the level of PEFR achieved. CONCLUSION: Anticholinergic agents such as ipratropium are not first-line treatments for acute asthma. They do not add any therapeutic benefit to the effects of albuterol given in divided doses over 1 hour, nor do they facilitate recovery in patients whose immediate response to sympathomimetics is impaired. PMID- 9209196 TI - Inapparent polycythemia vera: an unrecognized diagnosis. AB - PURPOSE: The Polycythemia Vera Study Group (PVSG) has established useful criteria for the diagnosis of polycythemia vera. In some circumstances, an increase of plasma volume (PV) masks that of red cell mass (RCM), with hemoglobin (Hb) and hematocrit (Ht) remaining normal. This defines the concept of inapparent polycythemia. PATIENTS AND METHODS: One hundred and three patients seen in the hematology unit with the diagnosis of polycythemia vera were studied. There were 55 males and 48 females with a median age of 59 years. Ninety-five patients fulfilled the PVSG criteria. Spontaneous erythroid colonies and low serum erythropoietin level confirmed the diagnosis in the 8 other cases. Patients were classified according to Hb and Ht level. RESULTS: Group A consisted of 85 patients with increased Hb and Ht defined, respectively, by Hb > 18 g/dl, Ht > 0.52 in males and Hb > 16 g/dL, Ht>0.47 in females. Group B included 18 patients (17%) with inapparent polycythemia vera (IPV) defined by a normal Hb and Ht value at diagnosis. In this group, the reasons to perform RCM were as follows: splenomegaly associated with increased platelets and/or leucocytes counts (n = 8), portal vein thrombosis (n = 5), increased platelets or leucocytes counts without splenomegaly (n = 3), and isolated splenomegaly (n = 2). The two groups were balanced in terms of age, sex, leucocyte, serum iron, and platelet level. Hemoglobin and Ht levels were significantly different between the two groups. The difference between the PV was indeed highly significant. The mean PV increase was + 9.5% (nL < +20%) in group A versus + 36.3% in group B (P < 0.00005). Red cell mass was not different between the two groups. CONCLUSIONS: Increased Hb or Ht should constitute the sole criteria for RCM determination. In the context of portal vein thrombosis, isolated hyperleucocytosis, thrombocytosis, or splenomegaly, a RCM should be performed. The frequency of IPV remains to be specified but the diagnosis of polycythemia vera is probably underestimated. PMID- 9209197 TI - Clinical correlates of elevated serum levels of interleukin 6 in patients with untreated Hodgkin's disease. AB - BACKGROUND: Interleukin-6 (IL-6) is a potent immunomodulatory cytokine that may have pathogenetic significance in several malignancies. In addition, high IL-6 levels have been associated with a poor prognosis in multiple myeloma, nonHodgkin's lymphoma, ovarian cancer, and renal cancer, as well in advanced Hodgkin's lymphoma. In this study, we analyzed IL-6 levels in newly diagnosed Hodgkin's disease and determined clinical correlates of elevated levels. PATIENTS AND METHODS: Using a sensitive enzyme-linked immunosorbent assay (lower limit of sensitivity = 0.35 pg/mL) we measured IL-6 levels in sera from 33 healthy controls and 65 untreated patients with Hodgkin's disease. RESULTS: Interleukin-6 levels in the Hodgkin's patients (median 2.7 pg/mL; range < 0.35 to 38.4 pg/mL) were significantly higher than in the controls (median < 0.35 pg/mL; range < 0.35 to 1.87 pg/mL; P < 0.0001). Interleukin-6 levels were also higher in males (P = 0.03) and in patients with bulky disease (P = 0.026) or advanced Ann Arbor stage (P = 0.017). In addition, serum levels of IL-6 also showed direct linear correlations with the erythrocyte sedimentation rate (r = 0.64, P = 0.0007), platelet count (r = 0.53, P < 0.0001), leukocyte count (r = 0.36, P = 0.003), and beta (2)-microglobulin level (r = 0.4, P = 0.0012); and an inverse linear correlation with serum albumin level (r = -0.43, P = 0.0003). In the 10 patients tested who had elevated serum IL-6 levels pretherapy and who achieved complete remission, serum IL-6 values decreased at the time of remission to the range found in healthy controls. CONCLUSIONS: Our observations suggest that, in patients with Hodgkin's disease, serum levels of IL-6 are frequently elevated at diagnosis, normalize during remission, and are associated with specific disease characteristics including several adverse prognostic features. PMID- 9209198 TI - Hormone replacement therapy in postmenopausal women: urinary N-telopeptide of type I collagen monitors therapeutic effect and predicts response of bone mineral density. AB - PURPOSE: To assess the ability of the urinary N-telopeptide of type I collagen (NTx) to monitor and predict therapeutic effects of hormone replacement therapy (HRT) in postmenopausal women. PATIENTS AND METHODS: To assess the relationship between baseline or change in NTx (predictive variable), and change in lumbar and hip bone mineral density (BMD; outcome variable), we conducted a 2-year randomized controlled study at academic university and private practice medical centers in 236 healthy women 1 to 3 years postmenopausal; 227 women completed the study. Women received estrogen plus progesterone plus calcium (treated group) or calcium alone (control group). RESULTS: In the treated group NTx significantly (P < 0.0001) decreased, and spine and hip BMD significantly (P < 0.00001 and P < 0.005, respectively) increased; in the control group NTx did not change but BMD decreased significantly (P < 0.01). Subjects in the highest quartiles for baseline NTx (67 to 188 units) or decreasing NTx (-66% to -87%) through 6 months demonstrated the greatest gain in BMD in response to HRT (P < 0.05 and P < 0.005). For every increase of 30 units in baseline NTx the odds of gain in BMD in response to HRT increased by a factor of 5.0 (95% confidence interval [CI] 1.9 to 13.3); for every 30% decrease in NTx through 6 months, the odds of gaining BMD in response to HRT increased by a factor of 2.6 (95% CI 1.6 to 4.4). In the control group an increase of 30 units in mean NTx across the study indicated a higher odds of losing BMD by a factor of 3.2 (95% CI 1.6 to 6.5). A high baseline NTx (> 67 units) indicated a 17.3 times higher risk of BMD loss if not treated with HRT. CONCLUSION: These data support the clinical utility of NTx to monitor the antiresorptive effect of HRT in recently postmenopausal women, and to predict changes in BMD in response to HRT. PMID- 9209199 TI - Metabolic control and prevalent cardiovascular disease in non-insulin-dependent diabetes mellitus (NIDDM): The NIDDM Patient Outcome Research Team. AB - PURPOSE: Cardiovascular disease is a major cause of morbidity and death in non insulin-dependent diabetes mellitus (NIDDM). While hyperglycemia is clearly related to diabetic microvascular complications, it contribution to large-vessel atherosclerosis is controversial. PATIENTS AND METHODS: We performed an analysis of the association between glycemic control and prevalent cardiovascular disease in 1,539 participants in the NIDDM Patient Outcomes Research Team study who were under usual care in a health maintenance organization. Prevalent cardiovascular disease and its risk factors were identified by self-administered questionnaire. Cardiovascular disease was defined by the presence of coronary heart disease, peripheral vascular disease, and/or cerebrovascular disease. Glycohemoglobin and lipid levels were obtained from a computerized laboratory database. RESULTS: The mean age of participants was 63 years (range 31 to 91); 51% were women. The mean duration of NIDDM was 9 years (range < 1 to 50), 35% took insulin, and 48% took sulfonylureas. Mean glycohemoglobin was 10.6%. Sixty percent had hypertension, 16% currently smoked cigarettes, and the mean total high-density lipoprotein (HDL) cholesterol ratio was 5.7. Fifty-one percent had cardiovascular disease. Cardiovascular disease prevalence remained constant across increasing quartiles of glycohemoglobin for both men and women. In contrast, prevalent cardiovascular disease was associated with established cardiovascular disease risk factors including age (67 versus 59 years, P < 0.0001), hypertension (66% versus 54%, P < 0.0001), current cigarette smoking (17% versus 13%, P < 0.005), and total/HDL cholesterol ratio (5.9 versus 5.6, P < 0.005). Cardiovascular disease was also associated with duration of NIDDM (11 versus 8 years, P < 0.0001). In multiple logistic regression analysis controlling for established cardiovascular disease risk factors and diabetes duration and therapy, glycohemoglobin remained unassociated with cardiovascular disease. CONCLUSIONS: Glycemic control is not associated with prevalent cardiovascular disease in this large population of individuals with NIDDM. Conventional cardiovascular disease risk factors are independently associated with cardiovascular disease and be a more promising focus for clinical intervention to reduce atherosclerotic complications in NIDDM. PMID- 9209200 TI - A more valid measurement of low-density lipoprotein cholesterol in diabetic patients. AB - PURPOSE: This study was conducted to determine if the direct low-density lipoprotein (LDL) cholesterol assay would provide a more valid measure of LDL cholesterol in diabetic patients compared with the Friedewald equation. PATIENTS AND METHODS: Fasting plasma from 148 diabetic patients, 40 with insulin-dependent diabetes (IDDM) and 108 with non-insulin dependent diabetes (NIDDM) with triglyceride levels < 400 mg/dL, were analyzed for LDL cholesterol using the Friedewald equation, the direct LDL assay, and beta-quantification. Forty-six diabetic patients with triglyceride levels > or = 400 mg/dL were also studied to determine the validity of the direct LDL cholesterol assay with hypertriglyceridemia. RESULTS: The friedewald equation and the direct LDL cholesterol assay correlated well with beta-quantification (r = 0.969 and r = 0.971, respectively) for LDL cholesterol determination in diabetic patients. Although the Friedewald equation in comparison with beta-quantification underestimated (8%) LDL cholesterol values in diabetic patients, the direct LDL cholesterol assay had a mean bias of < 1%. Also, the underestimation by the Friedewald equation exceeded 10% for the triglyceride subgroup of 200 to 400 mg/dL. Furthermore, the accuracy of the direct LDL cholesterol assay was superior to the Friedewald equation since LDL cholesterol levels determined by the two methods coincided within +/- 10% of beta-quantification in 85% and 68% of diabetic patients, respectively (P = 0.0005). Similar results for both the Friedewald equation and the direct LDL cholesterol assay in comparison with beta quantification were seen when diabetic patients were subgrouped into IDDM and NIDDM. Also, the direct LDL cholesterol assay appeared to provide a reliable estimate in patients with triglycerides > or = 400 mg/dL. CONCLUSION: The results of our studies indicate that the direct LDL cholesterol assay is a more reliable and accurate method than the Friedewald formula for LDL cholesterol determination in diabetic patients and is more rapid and cost effective than the reference method. PMID- 9209201 TI - How safe are serotonin reuptake inhibitors for depression in patients with coronary heart disease? AB - Depression occurs frequently in patients with coronary heart disease (CHD), and confers significant risk for additional morbidity and mortality. The cardiac effects of the tricyclic antidepressants (TCAs) have been well characterized. In contrast, the cardiac effects of the selective serotonin reuptake inhibitors (SSRIs) have been less thoroughly investigated. The Medline database from 1986 to 1996 was searched for all reports of cardiac effects of SSRIs, and this literature is summarized. In addition, potential drug interactions, reports of side effects, and efficacy studies in the elderly are reviewed. Finally, recommendations are made considering the risk/benefit ratio. PMID- 9209202 TI - Gabapentin for parkinsonism: a double-blind, placebo-controlled, crossover trial. AB - PURPOSE: Gabapentin is a recently available anticonvulsant whose mechanism of action remains unknown. We suspected efficacy from serendipitous observations of gabapentin in patients with parkinsonism. This led us to a double-blind, placebo controlled, crossover trial. PATIENTS AND METHODS: We administered gabapentin in a placebo-controlled, double-blind, crossover trial to 19 subjects with advanced parkinsonism. We measured the effect of placebo and gabapentin on subjects' symptoms with the Unified Parkinson's Disease Rating Scale, the Webster Scale, and the Hoehn and Yahr Scale. We assessed tremor with surface-recorded electromyography. RESULTS: Total Unified Parkinson's Disease Rating Scale improved with gabapentin compared with placebo (P = 0.0005). Likewise, activities of daily living and examination subscore of the Unified Parkinson's Disease Rating Scale improved with gabapentin compared with placebo but did not achieve statistical significance. Webster Scale showed improvement but neither Hoehn and Yahr Scale nor Webster Scale changes reached statistical significance. Tremor as measured by the Unified Parkinson's Disease Rating Scale improved with gabapentin but the use of the root mean square of the rectified electromyography as a measure of tremor activity was not statistically significant. CONCLUSIONS: This study demonstrates that gabapentin improves rigidity, bradykinesia, and tremor of parkinsonism including both Parkinson's disease and Parkinson's syndrome. The rigidity and bradykinesia of parkinsonism improve on the drug even when the effects of gabapentin on tremor are discounted. PMID- 9209203 TI - Epidemiology, pathophysiology, and management of hyponatremic encephalopathy. AB - Hyponatremia is the most common electrolyte abnormality among hospitalized patients. Death or brain damage associated with hyponatremia has been described since 1935, and it is now evident that hyponatremia can lead to death in otherwise healthy individuals. In the past, it had been assumed that the likelihood of brain damage from hyponatremia was directly related to either a rapid decline in plasma sodium or a particularly low level of plasma sodium. Recent studies have demonstrated that other factors may be more important. These factors include the age and gender of the individual, with children and menstruant women the most susceptible. Although many clinical settings are associated with hyponatremia, those most often associated with brain damage are postoperative, polydipsia, pharmacological agents, and heart failure. Morbidity and mortality associated with hyponatremia are primarily a result of brain edema, hypoxemia, and associated hormonal factors. Management of hyponatremia is largely determined by symptomatology. If the patient is asymptomatic, discontinuation of drugs plus water restriction is often sufficient. If the patient is symptomatic, active therapy to increase the plasma sodium with hypertonic NaCl is usually indicated. Although inappropriate therapy of hyponatremia can lead to brain damage, such an occurrence is rare. Thus, the risk of not treating a symptomatic patient for exceeds that of improper therapy. PMID- 9209204 TI - Prevention of recurrences of erosive reflux esophagitis: a cost-effectiveness analysis of maintenance proton pump inhibition. AB - PURPOSE: To determine the cost-effectiveness of three management strategies for healed erosive reflux esophagitis: maintenance therapy with a proton pump inhibitor (PPI) from the outset; no maintenance therapy unless a patient's symptoms recur once over a year; and no maintenance therapy unless a patient's symptoms recur twice over a year. MATERIALS AND METHODS: Decision analysis using data from randomized trials of lansoprazole, case series, and expert opinion. RESULTS: For patients with grade 4 esophagitis, maintenance from the outset is the most efficient approach. For all other patients, providing maintenance PPI after a patient experiences two recurrences is the least costly but least effective approach. The other two approaches prevent more recurrences: waiting to initiate maintenance therapy until symptoms recur once requires an additional $73 for each recurrence prevented whereas maintenance PPI from the outset requires an additional $819 for each recurrence prevented. Maintenance therapy from the outset is cost effective if symptoms of esophagitis cause a 22% or greater decrement in quality of life (using $50,000 per quality-adjusted life year gained as a cost-effectiveness definition). However, withholding maintenance until the time of a first recurrence is cost effective if symptoms cause a 2% or greater decrement in quality of life. CONCLUSION: For grades 2 and 3 esophagitis, providing maintenance therapy after a patient experiences a further recurrence is a preferred option that appears cost-effective across a wide array of assumptions. Maintenance therapy from the outset, however, appears cost-effective only for those patients who report a significant decline in quality of life associated with esophagitis or for those patients with baseline grade 4 esophagitis. PMID- 9209205 TI - Computerized decision support based on a clinical practice guideline improves compliance with care standards. AB - PURPOSE: Clinical guidelines are designed to assist in the management of specific diseases; however, these guidelines are often neglected in the delivery of care. The purpose of this study was to determine whether clinician use of an clinical practice guideline would increase in response to having, at the patient visit, a decision support system based on a practice guideline that generates a customized management protocol for the individual patient using data from the patient's electronic medical record. SUBJECTS AND METHODS: In a 6-month controlled trial at a primary care clinic, 58 primary care clinicians were randomized to receive either a special encounter form with the computer-generated guideline recommendations or a standard encounter form. The effect of computer-generated advice on clinician behavior was measured as rate of compliance with guideline recommendations. Data from 30 clinicians were analyzed; data from 28 clinicians were excluded because these clinicians did not meet predefined criteria for minimum exposure to diabetic patient care. RESULTS: Availability of patient management recommendations generated by the decision support system resulted in a two-fold increase in clinician compliance with care guidelines for diabetes mellitus (P = 0.01). Median compliance for the group receiving the recommendations was 32.0% versus 15.6% for the control group. CONCLUSION: Decision support based on a clinical practice guideline is an effective tool for assisting clinicians in the management of diabetic patients. This decision support system provides a model for how a clinical practice guideline can be integrated into the care process by computer to assist clinicians in managing a specific disease through helping them comply with care standards. Use of decision support systems based on clinical practice guidelines could ultimately improve the quality of medical care. PMID- 9209206 TI - An overview of metformin in the treatment of type 2 diabetes mellitus. AB - Type 2 diabetes mellitus results from impaired insulin secretion and reduced peripheral insulin sensitivity. Treatment options include diet, oral antihyperglycemic agents, and insulin. Metformin, an oral biguanide, ameliorates hyperglycemia by improving peripheral sensitivity to insulin, and reducing gastrointestinal glucose absorption and hepatic glucose production. Unlike sulfonylureas, it does not stimulate insulin secretion, aggravate hyperinsulinemia, or cause hypoglycemia or weight gain (weight stabilizes or decreases). It also has beneficial effects on serum lipid profiles. In lean or overweight type 2 diabetic patients uncontrolled by diet, metformin monotherapy significantly improves glycemic control, compared with placebo, and to similar extents as sulfonylurea monotherapy. In secondary sulfonylurea failure, combination metformin-sulfonylurea treatment significantly improves glycemic control beyond that achieved with either agent along. Metformin-sulfonylurea also appears to be as effective as insulin or insulin plus sulfonylurea, suggesting that such combination therapy may obviate or substantially delay insulin therapy. Limited data suggest that metformin-insulin therapy may improve glycemic control, possibly reducing insulin requirements, in type 2 diabetic patients uncontrolled by insulin alone following secondary sulfonylurea failure. Gastrointestinal side effects are common, but usually tolerated. Lactic acidosis risk is minimal, provided that contraindications, particularly renal impairment, and prescribing guidelines are respected. Aside from elevated plasma metformin levels with cimetidine and synergistic hypoglycemia with sulfonylureas, few interactions occur. Thus, metformin is safe and effective both as monotherapy or in combination with other antihyperglycemic agents in type 2 diabetic patients requiring additional glycemic control and may be advantageous when weight control is desirable and/or hyperlipidemia exists. PMID- 9209208 TI - Autoimmune-associated hemophagocytic syndrome. PMID- 9209207 TI - Adjunctive treatment of streptococcal toxic shock syndrome using intravenous immunoglobulin: case report and review. PMID- 9209209 TI - The civic hospital. PMID- 9209210 TI - Chest pain, congestive heart failure, and death in a 65-year-old man. PMID- 9209211 TI - Internal medicine residencies in managed-care era. PMID- 9209212 TI - Strategic business planning for internal medicine. PMID- 9209213 TI - Isoetherine or albuterol for acute asthma. PMID- 9209214 TI - The Nazi doctors. PMID- 9209215 TI - Sarcoidosis mortality in the United States. PMID- 9209216 TI - Secondary prophylaxis for visceral leishmaniasis in HIV-infected individuals. PMID- 9209217 TI - Description and evaluation of the vallecula sign: a new radiologic sign in the diagnosis of adult epiglottitis. AB - STUDY OBJECTIVE: To describe and prospectively evaluate a new radiologic sign with the potential to increase the diagnostic accuracy of soft-tissue radiography of the neck in the identification of adult epiglottitis. METHODS: We conducted a prospective, before-and-after blinded study at two tertiary care institutions. A convenience sample of four staff emergency physicians, three otolaryngology residents, four radiology residents, and four senior medical students volunteered to participate. We assembled 26 soft-tissue radiographs of the neck from consecutive patients ED with the diagnosis of epiglottitis made on the basic of direct visualization. Twenty-six control radiographs were identified from ED patients who were being evaluated for the presence of foreign bodies or minor cervical trauma. We then randomly mixed the two sets of radiographs. Participants were asked to identify epiglottis among the 52 randomly sequenced radiographs. A standardized 5-minute tutorial on the vallecula sign was presented to all participants after the first interpretation. We then asked the participants to make a second interpretation of the 52 radiographs without knowledge of correct answers from the initial evaluation. RESULTS: The participants accurately classified 80.5% of all radiographs reviewed before the tutorial and 98.8% after the tutorial (P < .0001). Similarly, sensitivity improved from 78.5% to 98.2% (P < .0001) and specificity improved from 82.8% to 99.5% (P < .0001). We found no significant differences in performance characteristics among the different types of participants. CONCLUSION: We have described a new radiographic sign that improves the diagnostic accuracy of soft-tissue radiography of the neck. If reproduced in prospective studies, the absence of the vallecula sign on radiography might obviate the need for routine use of direct visualization as an initial screen. PMID- 9209218 TI - Effect of oral contrast administration for abdominal computed tomography in the evaluation of acute blunt trauma. AB - STUDY OBJECTIVE: To determine how frequently oral contrast medium (OC) is essential for computed tomography (CT) diagnosis of blunt intraabdominal injury and to quantify the delay associated with OC administration and the incidence of adverse effects. METHODS: This retrospective chart review, with prospective reevaluation of CT scans for diagnostic value of OC, took place in a university teaching hospital and Level l trauma center. Participants were blunt-trauma victims admitted between June 1, 1988, and November 1, 1993, who had abdominal CT as part of their initial evaluation. Trauma registry records were used to identify study patients. Available charts and CTs were reviewed for all patients with intestinal/mesenteric and pancreatic injuries. Randomly selected cases of liver injury, spleen injury, and no intraabdominal injury were also reviewed. Blinded CT scans were reevaluated for quality of bowel opacification and value of OC to diagnostic impression. RESULTS: During the study period, 2,162 blunt-trauma patients had an abdominal CT; 297 intraabdominal injuries were diagnosed in 248 patients. Full review was done on 124 charts, and 70 CT scans were reevaluated. Thirty-one (100%) of 31 liver and spleen injuries were diagnosed on CT, and OC was considered essential in none of these studies. One (4.5%) of 22 intestinal and mesenteric injuries was seen on CT, but this was the only such injury treated nonoperatively. None of 21 surgically confirmed intestinal/mesenteric injuries was seen on CT. Free air or free OC was seen in none of 7 cases of intestinal perforation. OC was judged essential in none of 20 scans in patients without intraabdominal injury. On 2 scans. OC was considered essential for the radiographic diagnosis. One of these was a normal pancreas at exploration (radiographic false-positive result). The only pancreatic injury requiring specific surgical treatment was missed on CT. Twenty-one percent of patients required placement of nasogastric tube for contrast administration after failing oral administration, and 23% vomited OC. One of 124 had documented aspiration of OC. Average additional time incurred in the ED for administration of OC was 144 minutes. CONCLUSION: OC is rarely essential for CT diagnosis of intraabdominal injury. It may improve sensitivity for pancreatic injury, but it does not help identify injuries requiring surgical treatment. Even with OC, CT is insensitive for intestinal injury. Vomiting and aspiration are significant risks. Use of OC adds a significant amount of time to ED evaluation. Adverse effects of OC administration, in this setting, may outweigh its benefits. PMID- 9209219 TI - Variation in ED use of computed tomography for patients with minor head injury. AB - STUDY OBJECTIVE: To determine the frequency of utilization, yield for brain injury, incidence of missed injury, and variation in the use of computed tomography (CT) for ED patients with minor head injury. METHODS: This retrospective health records survey was conducted over a 12-month period in the EDs at seven Canadian teaching institutions. Included in this review were adult patients who sustained acute minor head injury, defined as witnessed loss of consciousness or amnesia and a Glasgow Coma Scale score of 13 or greater. Data were collected by research assistants who were trained to select cases and abstract data in a standardized fashion according to a resource manual. Subsequently, patient eligibility was reviewed by the study coordinator and principal investigator. RESULTS: Of the 1,699 patients seen, 521 (30.7%) were referred for CT, and 418 (79.8%) of these scans were negative for any type of brain injury. Overall, 105 (6.2%) of these patients sustained acute brain injury, including 9 (.5%) with an epidural hematoma Cochran's Q test for homogeneity demonstrated significant variation between the seven centers for rate of ordering CT (P < .0001), from a low of 15.9% to a high of 70.4%. All five cases of "missed" hematoma occurred at the institutions with the highest and third highest rates of CT use. After controlling for possible differences in case severity and patient characteristics at each hospital, logistic regression analysis revealed that five of seven hospitals were significantly associated with the use of CT (respected odds ratios [OR], .4, .5, .5, 3.2, and 4.7). Three of the centers (two with the highest ordering rates) showed significant heterogeneity in the ordering of CT among their attending staff physicians, from a low of 6.5% to a high of 80.0%. CONCLUSION: There was considerable variation among institutions and individual physicians in the ordering of CT for patients with minor head injury. Although emergency physicians were selective when ordering CT, the yield of radiography was very low at all hospitals. None of the cases of "missed" intracranial hematoma came from the lowest ordering institutions, indicating that patients may be managed safely with a selective approach to CT use. These findings suggest great potential for more standardized and efficient use of CT of the head, possibly through the use of a clinical decision rule. PMID- 9209220 TI - High-dose hydrocortisone hemisuccinate in scorpion envenomation. AB - STUDY OBJECTIVE: Scorpion envenomation is a common life-threatening hazard in tropical and subtropical countries. Standard treatment is not clearly defined. Many therapies, such as steroids, are prescribed without experimental justification. We sought to assess the efficacy of systematic administration of intravenous hydrocortisone hemisuccinate (50 mg/kg) in scorpion envenomation. METHODS: Six hundred consecutive envenomated patients older than 10 years who presented to the ED of a nonteaching secondary hospital in an area of Tunisia endemic for scorpion envenomation were randomly assigned to receive hydrocortisone hemisuccinate 50 mg/kg (n = 305) or placebo (n = 295) in addition to standard medical care. Patients in the two groups had similar clinical characteristics on initial clinical evaluation. Each was categorized as grade 1 (absence of systemic symptoms) or grade 2 (systemic symptoms of scorpion envenomation). Patients were treated in the ED for up to 4 hours or in the ICU, depending on clinical severity. Steroid and placebo groups were compared according to mortality rate, change of severity grade 4 hours after presentation and treatment, and duration of hospital stay. RESULTS: Distribution of patients with respect to severity grade was similar in the two groups at the 4-hour clinical evaluation. We detected no significant difference at the time of discharge between steroid-and placebo-treated patients with respect to mortality (one patient in each group) or duration of hospital stay. Extra costs incurred through steroid administration totaled US $989,000. CONCLUSION: Our findings do not support the use of intravenous high-dose steroids in scorpion-envenomated patients. The discontinuation of this practice would reduce costs substantially. PMID- 9209221 TI - Clinical presentation and outcome of brown recluse spider bite. AB - STUDY OBJECTIVE: To examine the clinical presentation and outcome of patients treated in the ED or toxicology clinic for suspected brown recluse spider bites. METHODS: We assembled a retrospective case of patients at a southeastern US university hospital. Our study group comprised 111 patients with suspected brown recluse spider bites treated during a 30-month period. Our main outcome measures were the need for skin grafting and the development of other complications. RESULTS: The mean age of our subjects was 34 +/- 17 years. Thirteen patients (12%) brought the spider to the hospital, 22 (20%) saw a spider at the time of the bite, and an exclusively clinical diagnosis was made in the remaining 76 (68%). Most wounds (59%) involved the leg. At the time of presentation, 81% had central discoloration and 37% necrosis. Sixteen patients (14%) were systemically ill, and 6 (5%) were admitted to the hospital. Most (86%) were treated with antibiotics. Dapsone was infrequently used (9%) and had usually been prescribed before the patient's presentation to our ED. Only three patients (3%; 95% confidence interval, 1% to 8%) required grafting. Mild hemolytic anemia developed in one patient, and another had mild hemolysis and a mild coagulopathy; neither patient was taking dapsone. No deaths or serious complications occurred in our study group. CONCLUSION: In our series, long-term outcome after brown recluse spider bite was good. Serious complications were rare, as was the need for skin grafting. Because the vast majority of bites heal with supportive care alone, aggressive medical therapy does not appear warranted. PMID- 9209222 TI - Affinity-purified, mixed monospecific crotalid antivenom ovine Fab for the treatment of crotalid venom poisoning. AB - SUBJECT OBJECTIVE: To test the efficacy and safety of a new antivenom, affinity purified, mixed monospecific crotalid antivenom ovine Fab, in human subjects with minimal or moderate crotalid envenomation. METHODS: We conducted a prospective multicenter clinical trial of 11 patients 10 years or older with progressive manifestations after mild to moderate crotalid snakebite. After giving their consent, subjects received four to eight vials of study drug and were then repeatedly examined over 48 hours and at 7 and 14 days after discharge. Each patient's clinical condition was evaluated serially with the use of a validated severity score, as well as on the basis of the investigator's assessment. RESULTS: In all 11 subjects to the antivenom was judged by the investigator to have had a beneficial response. The severity score for each patient remained the same or decreased over the first 4 hours. However, two subjects demonstrated worsened condition 12 to 15 hours after antivenom administration. In no subject did an allergic reaction develop. CONCLUSION: In this patient group, affinity purified, mixed monospecific crotalid antivenom ovine Fab was associated with a halt of progressive crotalid venom poisoning. Initial safety data are promising but must be addressed further in subsequent studies. PMID- 9209223 TI - Thrombocytopenia following timber rattlesnake envenomation. AB - STUDY OBJECTIVE: To better characterize timer rattlesnake venom--induced thrombocytopenia and coagulopathy and the response to therapy with Antivenin (Crotalidae) Polyvalent. METHODS: We conducted a retrospective multicenter review of timber rattlesnake envenomation. RESULTS: We reviewed 18 cases at two institutions. Restoration of normal prothrombin time and partial thromboplastin time was achieved in all cases with antivenom therapy. In contrast, complete reversal of thrombocytopenia was not achieved, despite antivenom therapy. CONCLUSION: Antivenom (Crotalidae) Polyvalent was less effective in reversing thrombocytopenia than coagulopathy after timber rattlesnake envenomation, suggesting that a component of timber rattlesnake venom persists in the blood despite antivenom therapy. Persistent thrombocytopenia may be due to a venom factor that the antivenom does not neutralize or to inadequate dosing of antivenom. Prompt reversal of thrombocytopenia following treatment of timber rattlesnake envenomation with this antivenom appears unlikely. PMID- 9209224 TI - Canebrake rattlesnake envenomation. AB - STUDY OBJECTIVE: To document the clinical presentation and course of consecutive cases of envenomation by the canebrake rattlesnake (Crotalus horridus atricaudatus). METHODS: The medical care provided all patients envenomated by canebrake rattlesnakes in two institutions was retrospectively reviewed. Particular attention was paid to coagulation abnormalities, serum enzymes, electrocardiographic findings, and treatment with antivenom. RESULTS: Of the 15 patients, envenomation was judged to be mild in 3, moderate in 6, and severe in 5. In one case envenomation caused death. The clinical course generally predicted the degree of increase in the serum creatinine kinase (CK) level, as well as the amount of antivenom used for treatment. Despite increases in serum CK concentration and frequent increases in the serum CK-MB fraction, we found no evidence of cardiac damage. CONCLUSION: In envenomation by North American pit vipers, rhabdomyolysis appears to be characteristic of envenomation by the canebrake rattlesnake. We speculate that toxins in the canebrake venom cause skeletal muscle rhabdomyolysis. Physicians caring for victims of canebrake bite should regard marked increases in CK concentration as coming from skeletal, not cardiac, muscle. Good general medical support and antivenom for victims with moderate to severe envenomation appears effective. PMID- 9209225 TI - Relationship of venom effects to venom antigen and antivenom serum concentrations in a patient with Crotalus atrox envenomation treated with a Fab antivenom. AB - STUDY OBJECTIVE: To describe the association among venom antigenemia, serum antivenom concentrations, and venom effects in a 53-year-old woman who was bitten by a Western Diamondback rattlesnake (Crotalus atrox). METHODS: The patient was enrolled in a multicenter trial of an investigational Fab antivenom. Her clinical condition and coagulation parameters were monitored for 2 weeks after the bite. RESULTS: After antivenom administration, the progression of the venom's effects was arrested. The antivenom reversed some local venom effects, caused venom antigens to disappear from the blood, and resolved the patient's profound thrombocytopenia (before antivenom, 12,000/mm3; 1 hour after antivenom, 227,000/mm3). Local venom effects recurred twice in the 24 hours after antivenom administration but were easily managed with additional Fab antivenom. Venom antigenemia was detected on days 5 and 8 after the initial treatment and was accompanied in one instance by the new onset of hypofibrinogenemia (119 mg/L) that resolved spontaneously and in both instances by renewed profound thrombocytopenia. Repeat Fab antivenom does no days 6 and 9 were followed by increases in platelet count (from 16,000 to 40,000/mm3 and from 11,000 to 20,000/mm3, respectively) and by the reduction or disappearance of venom antigenemia. The patient sustained no significant bleeding complications, and all laboratory values had returned to normal 2 weeks after the bite. CONCLUSION: Initial control of local symptoms and coagulopathy was prompt after the administration of Fab antivenom. Repeat doses during the 24 to 36 hours after a bite may be necessary for local control. Recrudescence of coagulopathy was likely due in part to renewed venom antigenemia after clearance of Fab antivenom. The role of Fab antivenom in the treatment of recurrent coagulopathy requires further study. PMID- 9209226 TI - Successful treatment of crotalid-induced neurotoxicity with a new polyspecific crotalid Fab antivenom. AB - STUDY OBJECTIVE: To report the effectiveness of a new polyvalent crotalid antivenom on neurotoxicity associated with North American rattlesnake envenomation. Two syndromes of crotalid-induced neurotoxicity have been reported. In severe envenomation by Crotalus scutulatus scutulatus (Mojave rattlesnake), weakness and fasciculations of various muscle groups, including those innervated by cranial nerves, may develop. Occasionally respiratory insufficiency develops. The second neurotoxic effects is myokymia, a type of fasciculation most frequently reported after bites by Crotalus horridus horridus (timber rattlesnake) and Crotalus atrox (Western diamondback rattlesnake). Conventional polyvalent antivenom is often ineffective in the treatment of venom-induced neurotoxicity. METHODS: We report a case series of three patients envenomated by North American rattlesnakes, one of which was identified as C scutulatus scutulatus. All three patients experienced neurotoxicity with weakness, paresthesias, and dramatic fasciculations, along with other signs and symptoms of crotalid venom poisoning. RESULTS: The administration of new polyspecific crotalid antivenom made of ovine Fab was successful in immediately and completely reversing neurotoxicity in each of these patients. CONCLUSION: We report the use of a new antivenom for North American crotalid envenomation that seems to have efficacy in reversing neurotoxicity associated with these bites. PMID- 9209227 TI - Association of rattlesnake bite location with severity of clinical manifestations. AB - STUDY OBJECTIVE: To examine an association between bite location in cases of North American crotalid envenomation and the severity of clinical manifestations. METHODS: We conducted a retrospective review of prospectively collected data for an experimental trial of crotalid antivenom. Our subjects were otherwise healthy individuals with minimal to moderate North American crotalid envenomation. We compared the severity of envenomation for patients with digit bites distal to the proximal interphalangeal joint and bites more proximal using a previously developed and validated snakebite severity score. RESULTS: Thirteen subjects were classified as having distal bites and 24 as having proximal bites. At baseline (before antivenom administration), the distal group had a mean severity score of 2.9 +/- 1.1, whereas the proximal group had a mean severity score of 5.0 +/- 2.2 (P = .0024). Patients in the proximal group tended to demonstrate a more rapid initial decline in severity score after receiving antivenom than did the distal group. CONCLUSION: In minimal to moderate North America crotalid envenomation, patients who sustained bites on distal aspects the digits tended to experience less severe clinical manifestations of envenomation. It is possible that an isolated bite to the distal aspect of a finger is an early marker of minimal envenomation. PMID- 9209228 TI - On-line medical control versus protocol-based prehospital care. AB - STUDY OBJECTIVE: To compare on-scene time, appropriateness of therapy, and accuracy of paramedic clinical assessments when prehospital care was provided with the use of on-line medical control (OLMC) by EMS-certified nurses from a single base station or by paramedics using chief complaint-based protocols. METHODS: We assembled a prospective before-and-after series to compare OLMC (phase 1) and protocol (phase 2) care rendered by all paramedics in a single urban municipality using a single base station. The subjects were consecutively enrolled patients who met protocol inclusion criteria and presented with altered level of consciousness, nontraumatic chest pain, or shortness of breath. For both phases, EMS and corresponding ED records were compiled; all references identifying phase were removed. After establishing interrater reliability, we randomly assigned charts to one of two reviewers for scoring. Complaint-specific scoring elements included on-scene time, assessments performed, presence or absence of indications for common treatments, treatments given, paramedic diagnosis, and emergency physician diagnosis. The percentages of inappropriate treatment decisions and paramedic diagnostic accuracy (versus that of the receiving emergency physician) were calculated. RESULTS: Phase 1 comprised 287 patients, phase 2 294. Interrater reliability between the two scorers was high. Of 2,190 elements scored jointly, the raters agreed in 97%, with kappa-values ranging from .6 to 1.0. On-scene time was 1 minute shorter during phase 2 (95% confidence interval [CI] for difference in median time, 0 to 2 minutes; P < .03). From phase 1 to phase 2 (relative risk [RR], 1.5; 95% CI, 1.0 to 2.1), inappropriate treatment decisions decreased from 7.4% to 5.1%. The percentage of cases in which paramedics and physicians were in complete diagnostic agreement was high (77% to 78%) and did not change across phases. CONCLUSION: The use of protocols resulted in small improvements in both on-scene time and the appropriateness of therapeutic decisions, without a change in agreement between paramedic and physician. Protocol care for these three chief complaints is clinically safe and, by reducing training and staffing considerations, may offer a cost-effective alternative to OLMC. PMID- 9209229 TI - Unwitnessed out-of-hospital cardiac arrest: is resuscitation worthwhile? AB - STUDY OBJECTIVE: To determine the epidemiology of unwitnessed out-of-hospital cardiac arrest and the factors associated with survival after resuscitation using the Utstein style data collection. METHODS: We conducted a prospective cohort study in a 525,000-population city served by a single EMS system comprising a tiered response with physicians in the field. We studied consecutive unwitnessed out-of-hospital cardiac arrests that occurred between January 1, 1994, and December 31, 1995. We determined survival from cardiac arrest to discharge from hospital and the factors associated with survival. RESULTS: Of the 809 patients for whom resuscitation was considered, 205 (25.3%) had sustained unwitnessed arrests. Cardiac origin of arrest was verified in 52% of cases. The most common noncardiac causes of arrest were trauma, intoxication, near-drowning, and hanging. In 150 patients (73.2%) the presenting rhythm was asystole, in 28 (13.6%) it was pulseless electrical activity, and in 27 (13.2%) it was ventricular fibrillation. Resuscitation was attempted in 162 cases, 59 (36.4%) of whom demonstrated return of spontaneous circulation; 45 (27.8%) were hospitalized alive, and 8 (4.9%) were discharged. The survivors represented 6.7% of all out-of hospital cardiac arrest survivors during the study period. Survival was most likely if patients presented with pulseless electrical activity; none of the patients with asystole of cardiac origin survived. Sex (P = .032), age (inverse relationship, P = .0004), scene of collapse (P = .042), and interval from call receipt to arrival of first responders (P = .004) were associated with survival. In a logistic-regression model, near-drowning remained an independent factor of survival (odds ratio, 15.5; 95% confidence interval, 1.2 to 200). A routine priority dispatching protocol differentiated cardiac arrest patients with survival potential from those who already had irreversible signs of death. CONCLUSION: This survey shows that survival after unwitnessed out-of-hospital cardiac arrest is unlikely with an initial response of basic life support alone. Withdrawal of resuscitation should be considered if an adult victim of unwitnessed cardiac arrest is found in asystole and the arrest is of obvious cardiac origin. PMID- 9209231 TI - EMS-witnessed arrest and the evaluation of the effectiveness of care. PMID- 9209230 TI - Cardiac arrest witnessed by prehospital personnel: intersystem variation in initial rhythm as a basis for a proposed extension of the Utstein recommendations. AB - STUDY OBJECTIVE: To test the hypothesis that intersystem variation in initial rhythm among EMS-witnessed arrests is of sufficient magnitude to warrant standardization of survival by creation of an Utstein-style denominator of EMS witnessed ventricular fibrillation (VF). METHODS: We conducted a planned subset analysis of a prospective observational cohort study of consecutive EMS-witnessed adult cardiac arrests occurring in New York City and meeting Utstein entry criteria. The primary outcome measure was intersystem variation in frequency of EMS-witnessed VF in New York City compared with that in other EMS systems. Secondary outcome measures were variations in survival after EMS-witnessed VF arrests and overall survival after all EMS-witnessed arrests. RESULTS: Intersystem variation showed a threefold difference in the frequency of EMS witnessed VF (24% in New York City versus 77% in Scotland; 99% confidence interval [CI] for 53% difference, 43% to 63%; P < 10(-7), a twofold difference in survival after EMS-witnessed VF (25% in NYC versus 48% in King County, WA; 99% CI for 23% difference, 6% to 39%; P < .002), and a fourfold difference in survival after all EMS-witnessed arrests (9% in New York City versus 35% in King County; 99% CI for 26% difference, 18% to 34%; P < 10(-7). CONCLUSION: The marked variation in frequency of initial rhythm in EMS-witnessed arrests suggests that a modified Utstein denominator of EMS-witnessed VF would facilitate more uniform intersystem comparison of survival in this unique cohort. However, even after adjustment for initial rhythm, large residual intersystem survival differences remain unexplained. PMID- 9209232 TI - The role of emergency medical services in primary injury prevention. Consensus workshop. Arlington, Virginia, August 25-26, 1995. AB - Injury is a leading cause of death and disability. Preventing injuries from ever occurring is primary injury prevention (PIP). The objective of this statement is to present the consensus of a 16-member panel of leaders from the out-of-hospital emergency medical services (EMS) community on essential and desirable EMS PIP activities. Essential PIP activities for leaders and decision makers of every EMS system include: protecting individual EMS providers from injury; providing education to EMS providers in PIP fundamentals; supporting and promoting the collection and utilization of injury data; obtaining support for PIP activities; networking with other injury prevention organizations; empowering individual EMS providers to conduct PIP activities; interacting with the media to promote injury prevention; and participating in community injury prevention interventions. Essential PIP knowledge areas for EMS providers include: PIP principles; personal injury prevention and role modeling; safe emergency vehicle operation; injury risk identification; documentation of injury data; and one-on-one safety education. PMID- 9209233 TI - Panic disorder: diagnosis and treatment in emergency medicine. AB - Panic disorder is a newly recognized psychiatric illness involving unexpected, unprovoked attacks of anxiety. Patients with panic disorder commonly seek treatment in the ED. It is important for the emergency physician to properly recognize and categorize this disorder to initiate appropriate treatment. Illness description and treatment guidelines of this disorder are discussed in this article. PMID- 9209234 TI - Tylenol Extended Relief overdose. AB - In this report we describe the toxicokinetics of the Tylenol Extended Relief (TER) preparation of acetaminophen in human overdose. We collected 41 cases of TER overdose from five regional poison centers. Patients who met the following criteria were studied: a single ingestion of TER alone; confirmed time of ingestion; at least four acetaminophen determinations; and normal concentrations of liver function enzymes. With the exception of standard decontamination measures, treatment with N-acetylcysteine (NAC) if any acetaminophen level was above the treatment line of the Rumack-Matthew nomogram, and additional acetaminophen determinations, no interventions were recommended. Our study group comprised 13 patients, 12 female and 1 male, with single overdoses of 10.4 to 65 g TER. The acetaminophen elimination half-life was 3.1 +/- .8 hours (mean +/- SD; range, 1.3 to 4.0 hours; n = 12). The elimination phase for patients 2, 3, 4, 6, 8, 9, 11, 13 was delayed until 8.0 +/- 2.8 hours (range, 5 to 14 hours) after ingestion. Patients 3, 8, and 11--who had initial acetaminophen levels below the "possible toxicity" line of the Rumack-Matthew nomogram--later had acetaminophen levels above this line. No patient demonstrated a late or second acetaminophen peak. We conclude that the elimination half-life of TER acetaminophen is similar to that reported in overdose of immediate-release acetaminophen overdose. In a subgroup of patients, drug absorption continued beyond the 2 to 4 hours previously reported in immediate-release acetaminophen overdose. On the basis of our data, the use of a single 4-hour acetaminophen determination may lead to failure to recognize patients with potentially toxic TER ingestion. Until more toxicokinetic data are available, a reasonable approach would be to obtain at least one additional acetaminophen determination at least 4 to 6 hours after the first, if the first is obtained 4 to 8 hours after ingestion. NAC treatment should be initiated if either level is above the nomogram line but not if both levels fall below the nomogram line. PMID- 9209235 TI - Nontraumatic gas gangrene. AB - A 77-year-old man with a history significant only for coronary artery disease presented to the ED with left-arm pain, shortness of breath, nausea, and diaphoresis. Six hours after the patient's admission to the hospital for presumed unstable angina, fever and left arm swelling, associated with crepitus and violaceous bullae, developed. The patient was taken to the operating room, where he was found to have extensive myonecrosis requiring forequarter amputation of the left arm. Nontraumatic clostridial myonecrosis is a fulminant, often fatal infection. This rare condition is usually caused by Clostridium septicum and has a high association with underlying malignancy. The patient reported here was found to have a colonic lesion and acute leukemia, both previously undiagnosed. This case illustrates the insidious manner in which spontaneous myonecrosis may present. PMID- 9209237 TI - Gauging urgency. PMID- 9209238 TI - False-positive end-tidal CO2. PMID- 9209239 TI - Cadavers as teachers. PMID- 9209240 TI - Role of emergency physicians in the prevention of pediatric injury. American College of Emergency Physicians. PMID- 9209241 TI - Civil commitment. American College of Emergency Physicians. PMID- 9209242 TI - Guidelines concerning work stoppages and slowdowns. American College of Emergency Physicians. PMID- 9209243 TI - Olestra: assessing its potential to interact with drugs in the gastrointestinal tract. PMID- 9209244 TI - Influence of the CYP1A2 inhibitor fluvoxamine on tacrine pharmacokinetics in humans. AB - OBJECTIVE: Tacrine is extensively metabolized by cytochrome P4501A2 (CYP1A2). Fluvoxamine, a potent CYP1A2 inhibitor, may be coadministered with tacrine. The aim of this study was to examine the influence of fluvoxamine administration on the disposition kinetics of single-dose tacrine administration. METHODS: Thirteen healthy volunteers participated in this double-blind, randomized crossover study, which compared the effects of fluvoxamine (100 mg/day during 6 days) and placebo on the pharmacokinetics of a single oral dose of tacrine (40 mg). RESULTS: Fluvoxamine caused a significant increase in tacrine area under the plasma concentration versus time curve (AUC): arythmetic mean, 27 (95% confidence interval [CI], 19 to 38) ng.hr/ml versus 224 (95% CI, 166 to 302) ng. hr/ml. Fluvoxamine caused a decrease in the apparent oral clearance of tacrine from 1683 +/- 802 to 200 +/- 106 L/hr (mean +/- SD), which was explained by a decrease in its nonrenal clearance. Five subjects had gastrointestinal side effects during fluvoxamine administration. Fluvoxamine administration was associated with significant increases in the plasma AUC values of three monohydroxylated tacrine metabolites and in the total urinary recovery measurements of tacrine and its metabolites (9.1% +/- 4.6% versus 24.0% +/- 2.6% of recovery). These results may be attributable to fluvoxamine-dependent inhibition of CYP1A/, which is responsible of the biotransformation of tacrine into its monohydroxylated metabolites and further into dihydroxylated and reactive metabolites. CONCLUSION: Fluvoxamine inhibits the metabolism of tacrine. CYP1A2 may be the target of this inhibition. Fluvoxamine may modulate the hepatotoxicity of tacrine, depending on the relative contribution of tacrine and its reactive metabolites to this toxicity. PMID- 9209245 TI - Population pharmacokinetics of intravenous caffeine in neonates with apnea of prematurity. AB - OBJECTIVE: The study the population pharmacokinetics of caffeine after intravenous administration to premature infants with apnea. METHODS: A prospective, blinded parallel study in which daily caffeine citrate doses of 30, 15, and 3 mg/kg were administered over 7 days by intermittent intravenous infusion. Arterial blood samples (three to six per patient) were assayed for caffeine content by means of HPLC. Population pharmacokinetic modeling was performed with NONMEM. RESULTS: Clearance (L/hr) = (0.00000399 . current weight [grams]) + (0.000128 . postnatal age [days]). For gestational age > 28 weeks, volume of distribution (L) = (0.000764 . weight [grams] + (0.0468 . postnatal age [days]); for gestational age < or = 28 weeks, volume of distribution (L) = (0.000755 . weight [grams]) + (0.0224. postnatal age [days]). Interpatient variability (coefficient of variation, in percent) was approximately 25% for clearance and approximately 11% for volume of distribution. Intrapatient error (standard deviation) was 3.9 mg/L. There was insignificant bias between observed and model-predicated serum caffeine concentrations in a separate group of 30 infants. CONCLUSIONS: Caffeine was well tolerated at all doses. Clearance was markedly lower and volume of distribution was higher than the values reported previously for term infants and adults. Both parameters were significantly influenced by postnatal age and current body weight, whereas volume of distribution in infants > 28 weeks' gestational age was higher than that in more premature babies. The predictive performance and the clinical application of the derived population models was satisfactorily shown. PMID- 9209246 TI - Disposition of oral azithromycin in humans. AB - BACKGROUND: The oral bioavailability of azithromycin is approximately 37% in healthy subjects; little is known about the disposition of the remaining 63% of the dose. This study attempted to describe the fate of azithromycin before absorption. METHODS: Twelve subjects with ileostomies in place for > 1 month were studied in this open-label, randomized, three-center, two-period, two-treatment crossover study. Subjects randomly received single 500 mg intravenous infusion (over 1 hour) or two 250 mg oral capsules after a fast for > 12 hours. Blood and ileostomy samples were collected serially after each administration and analyzed for azithromycin and two metabolites (descladinose and 9a-N-desmethyl metabolites) by HPLC with electrochemical detection. RESULTS: Mean +/- SD peak concentration values after oral and intravenous administration were 0.21 +/- 0.08 and 3.40 +/- 1.12 microgram/ml. Mean values for area under the serum concentration versus time curve were 1.27 +/- 0.65 and 7.14 +/- 1.34 micrograms x hr/ml, respectively. The absolute bioavailability of 16.2% was approximately one half the value observed previously in healthy subjects. Recovery in ileostomy fluid (percent of dose in 24 hours) or azithromycin, descladinose, and 9a-N desmethyl metabolites were 13%, 0.5%, and 1% (total, 15%) after intravenous dosing and 47%, 13%, and 2% (total, 62%) after oral dosing. Total and ileal clearances were 776 +/- 126 and 158 +/- 63 ml/min after intravenous dosing. CONCLUSION: Because more descladinose metabolite was detected after oral dosing, acid degradation of azithromycin before absorption contributed to some loss in oral bioavailability. Further, ileal clearance (biliary plus intestinal excretion clearance) in this population represented 21% of total clearance. Taken together, these data suggest that the cause of low oral bioavailability of azithromycin is the result of incomplete absorption rather than acid degradation or extensive first-pass metabolism. PMID- 9209247 TI - Cotinine effects on nicotine metabolism. AB - BACKGROUND: Nicotine clearance and half-life are known to be significantly reduced in smokers compared to nonsmokers. Cotinine is the major primary metabolite of nicotine, and it accumulates in the body with regular smoking. Nicotine and cotinine appear to be metabolized by the same liver enzyme. Therefore we hypothesized that cotinine inhibits nicotine metabolism, resulting in slower nicotine clearance in smokers compared with nonsmokers. METHODS: This was a crossover, randomized, double-blind, and placebo-controlled study. The subjects were 12 healthy nonsmoking volunteers. They received two intravenous infusions of deuterium-labeled nicotine-d2 and cotinine-d4 (0.5 micrograms/kg/min), once with oral cotinine treatment of 0.25 mg/kg twice a day and once with placebo. Nicotine and cotinine pharmacokinetic parameters were determined for each infusion. RESULTS: During oral cotinine treatment, average plasma levels of cotinine ware 900 ng/ml, comparable to levels observed in some very heavy smokers. Cotinine had no effect on the disposition kinetics of nicotine-d2. The half-life of cotinine after low-dose cotinine-d4 infusion was comparable to that after high-dose cotinine described in previous studies. The half-life of labeled cotinine derived from nicotine was significantly longer than the half-life of cotinine administered as cotinine. CONCLUSIONS: Cotinine is not responsible for the lower nicotine clearance observed in smokers. Our data suggest that the pharmacokinetics of low-dose cotinine in nonsmokers do not differ from those of high-dose in smokers, and therefore cotinine levels can be used quantitatively in environmental tobacco exposure. The longer half-life of cotinine derived from nicotine suggests that slow release of nicotine from tissues is responsible for the apparent long half-life of cotinine in nonsmokers exposed to environmental tobacco smoke. PMID- 9209248 TI - Pharmacokinetics and ventilatory effects of oxycodone before and after liver transplantation. AB - The pharmacokinetics and ventilatory effects of oxycodone were studied in six volunteer patients with end-stage liver cirrhosis before and after orthotopic liver transplantation. Plasma samples and urine were collected for 24 hours after intravenous administration of 0.05 mg/kg oxycodone hydrochloride. Concentrations of oxycodone and its metabolites, noroxycodone and oxymorphone, were measured in plasma and urine. THe median elimination half-life of oxycodone was 13.9 hours (range, 4.6 to 24.4 hours) in patients with cirrhosis before transplantation and 3.4 hours (range, 2.6 to 5.1 hours) after transplantation (p < 0.05). Correspondingly, oxycodone clearance increased from 0.26 L/min (range, 0.15 to 0.73 L/min) before transplantation to 1.13 L/min (range, 0.71 to 3.98 L/min) after transplantation (p < 0.05). Oxycodone depressed ventilation more strongly before transplantation than after transplantation (p < 0.05). Care should be exercised when oxycodone is used in patients with end-stage disease. PMID- 9209249 TI - Coadministration of glyburide and minoxidil, drugs with opposing effects on potassium channels. AB - INTRODUCTION: Adenosine triphosphate (ATP)-sensitive potassium (K+) channels are modulated by drugs, so that they are opened by vasodilators such as minoxidil but are closed by hypoglycemic agents such as glyburide (glibenclamide). Animal studies and in vitro evidence suggests that the coadministration of drugs with opposing effects on K+ channels attenuates their pharmacodynamic effects. METHODS: To investigate whether this important pharmacodynamic interaction occurs in humans, we administered 5 mg minoxidil, 2.5 mg glyburide or both in a double blind fashion to nine healthy subjects. Glucose and insulin responses during an intravenous glucose tolerance test (0.3 gm/kg) were measured and blood pressure was recorded for 8 hours. In an additional four subjects the effect of 5 mg glyburide on the hypotensive effect of 5 mg minoxidil was examined. RESULTS: None of the parameters of glucose metabolism differed significantly when subjects received glyburide alone, minoxidil alone, or glyburide with minoxidil. Minoxidil or minoxidil in combination with 2.5 mg glyburide resulted in a similar significant decrease in blood pressure compared with the response to glyburide alone. The hypotensive effect of minoxidil was smaller in the four subjects who received the higher dose of glyburide, but significant hypoglycemia (blood glucose concentration < 60 mg/dl) occurred in three of the four subjects. CONCLUSION: We conclude that, in healthy volunteers, the coadministration of 2.5 mg glyburide and 5 mg minoxidil does not result in attenuation of the blood pressure-lowering effect of minoxidil. The smaller hypotensive response in four subjects who received 5 mg glyburide and 5 mg minoxidil suggests the possibility of a dose-related drug interaction. Studies with strict clamping of blood glucose concentrations will be required to address this possibility. PMID- 9209250 TI - Fenoterol but not dobutamine increases erythropoietin production in humans. AB - OBJECTIVE: This study assessed the role of adrenergic signal transmission in the control of renal erythropoietin (EPO) production in humans. METHODS: Forty-six healthy male volunteers underwent a hemorrhage of 750 ml. After phlebotomy, they received (intravenously for 6 hours in a parallel, randomized, placebo-controlled and single-blind design) either placebo (0.9% sodium chloride), or the beta 2 adrenergic receptor agonist fenoterol (1.5 microgram/min), or the beta 1 adrenergic receptor agonist dobutamine (5 micrograms/kg/min), or the nonselective beta-adrenergic receptor antagonist propranolol (loading dose of 0.14 mg/kg over 20 minutes, followed by 0.63 micrograms/kg/min). RESULTS: The AUCEPO(0-48 hr)fenoterol was 37% higher (p < 0.03) than AUCEPO(0-48 hr)placebo, whereas AUCEPO(0-48 hr)dobutamine and AUCEPO(0-48 hr)propranolol were comparable with placebo. Creatinine clearance was significantly increased during dobutamine treatment. Urinary cyclic adenosine monophosphate excretion was increased only by fenoterol treatment, whereas serum potassium levels were decreased. Plasma renin activity was significantly increased during dobutamine and fenoterol infusion. CONCLUSIONS: This study shows in a model of controlled, physiologic stimulation of renal erythropoietin production that the beta 2-adrenergic receptor agonist fenoterol but not the beta 1-adrenergic receptor agonist dobutamine is able to increase erythropoietin levels in humans. The result can be interpreted as a hint that signals for the control of erythropoietin production may be mediated by beta 2-adrenergic receptors rather than by beta 1-adrenergic receptors. It appears to be unlikely that an increase of renin concentrations or glomerular filtration rate is causally linked to the control of erythropoietin production in this experimental setting. PMID- 9209251 TI - Chloroquine-induced venodilation in human hand veins. AB - OBJECTIVE: Hypotension induced by parenteral administration of chloroquine is a common and serious adverse effect of this drug. Our aim was to investigate whether chloroquine produces venodilation in vivo and to explore the underlying mechanisms. METHODS: Vascular effects of chloroquine were studied in healthy volunteers with use of the dorsal hand vein technique at the Geriatric Research Education and Clinical Center, Veterans Affairs Palo Alto Health Care System. We studied 22 healthy volunteers (19 men and three women). Venous responsiveness was determined with the dorsal hand vein technique, which measures the diameter of the vein. RESULTS: Chloroquine was found to produce a dose-dependent relaxation of hand veins preconstricted with the alpha 1-receptor selective agonist phenylephrine. The venodilatory response to chloroquine ranged from 15% +/- 19% at an infusion rate of 0.75 microgram/min to 61% +/- 24% at 48 microgram/min. Venodilation was attenuated by the nitric-oxide synthase inhibitor NG-monomethyl L-arginine (L-NMMA) so that the dose of chloroquine required to produce 20% venodilation was increased from 3.7 micrograms/min to 15 micrograms/min (p < 0.01). In the presence of a combination of histamine receptor antagonists, there was also a diminution of the vasodilatory response to chloroquine from 72% +/- 5% to 44% +/- 5% at the infusion rate of 96 micrograms/min. The response was further reduced to 33% +/- 7% by the coinfusion of H1-/H2-receptor antagonists with L NMMA. CONCLUSION: Chloroquine produces venodilation at infusion rates that achieve local concentrations likely similar to those observed systemically after clinically relevant intravenous doses. The date also suggest a role for nitric oxide and histamine release in mediating this response. PMID- 9209252 TI - Platelet alpha 2-adrenergic receptors in hypercholesterolemia: relationship between binding studies and epinephrine-induced platelet aggregation. AB - BACKGROUND: Platelets isolated from patients with hypercholesterolemia are more sensitive in vitro to various aggregating agents, including epinephrine, than those isolated from normocholesterolemic subjects. Increased platelet reactivity is one mechanism that may explain the enhanced risk of thromboembolism in hypercholesterolemia. This study assessed whether platelet hyperreactivity to epinephrine in hypercholesterolemia is associated with higher alpha 2-adrenergic receptor density or affinity for epinephrine. METHODS: Platelet aggregation and binding studies, with use of [3H]yohimbine as ligand, were performed on platelets isolated from 30 patients with type IIa hypercholesterolemia and 23 control subjects. RESULTS: Platelet aggregation in response to epinephrine was significantly higher in patients with hypercholesterolemia than in control subjects. A statistically significantly higher alpha 2-adrenergic receptor density was observed in a subgroup of 13 patients with hypercholesterolemia than in 13 sex- and age-matched control subjects (280 +/- 61 and 230 +/- 49 fmol/mg protein respectively; p < 0.03), but no difference was observed in receptor affinity for the ligand. In these subgroups plasma total and levels of low density lipoprotein (LDL) cholesterol were inversely correlated with platelet aggregation but directly correlated with platelet receptor density. CONCLUSION: Platelet alpha 2-adrenergic receptor density is increased in hypercholesterolemia and directly correlates with plasma total and levels of LDL cholesterol, providing at least a partial explanation for the enhanced platelet response to epinephrine that is observed in hypercholesterolemia. PMID- 9209253 TI - Delayed sequelae after acute overdoses or poisonings: cranial neuropathy related to ethylene glycol ingestion. AB - A 31-year-old woman came to the hospital with breathlessness, confusion, and a refractory anion gap metabolic acidosis; acute renal failure subsequently developed. Her blood ethylene glycol concentration was 390 mg/L, and she was treated with an intravenous ethanol infusion and hemodialysis. During the tenth and eleventh day after admission bilateral seventh cranial nerve paralysis developed, as well as bilateral dysfunction of cranial nerves II, V, VIII, IX, X and XII. Magnetic resonance imaging of her head showed gadolinium enhancement of the fifth cranial nerve bilaterally and a communicating hydrocephalus. Over the subsequent 11 months she recovered full function of her cranial nerves V, VII, IX, X, and XII, and she had subjective clinical improvement to baseline function in cranial nerves II and VIII. This case serves to introduce a discussion of agents that cause delayed complications after their acute toxic ingestion. PMID- 9209254 TI - Race and angioedema. PMID- 9209255 TI - Alumni self-perception of competence at time of dental school graduation. AB - A survey instrument was administered to recent alumni to evaluate self-perceived competency in twenty-one selected areas at the time of graduation. The questionnaire was completed by 439 (62.8 percent) alumni who graduated between the years 1985 and 1994, inclusive. Graduates generally felt most competent in their ability to treat dental caries and its sequelae and least competent in their ability to recognize myofascial pain and temporomandibular joint disorders. Very little association was demonstrated between demographic characteristics of the respondents and their competency self-ratings. The survey identified areas of strengths and weakness within the present comprehensive care curriculum as perceived by the graduating dentists. PMID- 9209258 TI - Characteristics identified by deans as essential to success. PMID- 9209256 TI - Assessment of preclinical problem-based learning versus lecture-based learning. AB - Academic performance on a standardized oral comprehensive exam (OCE) was compared for students taught basic science in a problem-based learning (PBL) curriculum and a lecture-based learning (LBL) curriculum. The OCE was administered to the graduating classes of 1991-1994 (n approximately 20/class) six months after completion of their basic science courses. The OCE contained six components including: Organization and Thoroughness, Diagnosis, Primary Treatment Plan, Alternate Treatment Plan, Science and Medical Knowledge, and Dental Knowledge. Six to eight examiners graded each of the students by using a standardized scoring system and by subjective comments. The class of 1991 was taught by LBL, classes of 1993 and 1994 by PBL, and the class of 1992 by an incomplete PBL teaching method. Mean OCE scores were not significantly different between classes; however, the Science and Medical Knowledge component score was significantly better for the class of 1994 than for 1991 (p < 0.05). There was a non-significant 40 percent increase (p = 0.07) in honors and a 269 percent (p < 0.001) increase in cumulative positive examiner comments between 1991 and 1994. PMID- 9209259 TI - Pediatric dentistry faculty profile and plans to remain active. PMID- 9209260 TI - The application of computers to accreditation. PMID- 9209257 TI - The effect of cooperative learning strategy on preclinical performance. PMID- 9209261 TI - Comments concerning graduate medical education. PMID- 9209262 TI - Extensive peroxynitrite activity during progressive stages of central nervous system inflammation. AB - Nitric oxide (NO) production has been associated with disease activity in multiple sclerosis and experimental autoimmune encephalomyelitis (EAE). This free radical can be transformed by superoxide to peroxynitrite, an extremely toxic oxidant which causes lipid peroxidation. In addition, peroxynitrite nitrates tyrosine residues, resulting in nitrotyrosine, which can be identified immunohistochemically. The results of this study indicate that peroxynitrite is formed very early during EAE development, correlating with clinical disease activity. Nitrotyrosine-positive cells display a widespread distribution in brain and spinal cord during severe disease and are associated with both perivascular infiltrates and parenchymal sites. Double-staining procedures demonstrated that a subpopulation of CD11b-positive cells (macrophages/microglia) reacted with nitrotyrosine antibodies. Immunostaining for inducible NO synthase demonstrated a similar distribution as nitrotyrosine staining. These experiments indicate that peroxynitrite is formed during progressive stages of disease activity. PMID- 9209263 TI - Lymphocyte-specific inducible expression of potassium channel beta subunits. AB - Many studies have shown that voltage-gated potassium (Kv) channel activity is essential for T-lymphocyte proliferation. The IL-2-inducible neuroimmune gene, I2rf5 is the mouse homologue of the rat Kv beta 2 subunit. In this study we show that in addition to constitutive expression in adult murine brain, expression of Kv channel subunits beta 1.1 and beta 2.1 is inducible in a cloned T-helper cell line stimulated with IL-2 and in normal murine splenocytes stimulated with Con A or LPS. This expression pattern appears to be lymphocyte specific, because stimulated fibroblasts and vascular smooth muscle cells do not express Kv beta channel subunit mRNA. These observations suggest that Kv beta subunit expression is tissue specific and inducible in stimulated lymphocytes. Because Kv beta subunits modulate K+ channel activity, the inducible and variable expression of these subunits in lymphocytes may represent an additional regulatory mechanism for lymphocyte proliferation. PMID- 9209264 TI - Identification of cell types producing RANTES, MIP-1 alpha and MIP-1 beta in rat experimental autoimmune encephalomyelitis by in situ hybridization. AB - The chemokines RANTES, macrophage inflammatory protein (MIP)-1 alpha and MIP-1 beta are members of the beta-family of chemokines and potent chemoattractants for lymphocytes and monocytes. To investigate the factors which regulate lymphocyte traffic in experimental autoimmune encephalomyelitis (EAE), we studied, by in situ hybridization analysis, the kinetics of mRNA expression and the potent cellular sources of RANTES, MIP-1 alpha and MIP-1 beta in the central nervous system (CNS) during the course of EAE. RANTES-positive cells appeared in the subarachnoid space and infiltrated the subpial region at around day 10, increased to a peak at days 12-13 and then decreased following the resolution of the acute phase of EAE, though elevated RANTES message expressions still remained on chronic subclinical stage. Most of RANTES positive cells were identified as T lymphocytes located mainly around blood vessels, by combined studies of in situ hybridization and immunohistochemistry. The remainder of the RANTES-positive cells were astrocytes and macrophages/microglia. MIP-1 alpha and MIP-1 beta mRNA positive cells appeared around day 10, increased further on days 12-13 and then gradually decreased. Most of the MIP-1 alpha- and MIP-1 beta-positive mononuclear cells were located around blood vessels. The kinetics of RANTES, MIP-1 alpha and MIP-1 beta expression paralleled those of the recruitment of infiltrating inflammatory cells and disease severity. Our observations support the possibility that chemokine production by T-cells, macrophages and astrocytes lead to the infiltration of inflammatory cells into the CNS parenchyma during the acute phase of EAE. PMID- 9209265 TI - Analysis of a sequenced cDNA library from multiple sclerosis lesions. AB - To identify genes that are expressed in MS pathogenesis, we have analyzed a normalized cDNA library made from mRNA obtained from CNS lesions of a patient with primary progressive MS. Complementary DNA clones obtained from this library were subjected to automated DNA sequencing to generate expressed sequence tags. Analysis of this MS cDNA library revealed the presence of 54 cDNAs that were associated with immune activation and indicated the presence of an ongoing inflammatory response with evidence of both cell-mediated and humoral immune responses. The surprising finding was that 16 of the cDNAs encoded autoantigens associated with seven other autoimmune disorders, while only three of these 16 autoantigen cDNAs were present in a similarly constructed adult brain library. Such aberrant autoantigen expression could provide a source of secondary autoimmune stimulation that could contribute to the ongoing inflammatory response in MS. In addition, two cDNAs were found that mapped to a known MS susceptibility locus (5p14-p12): one encoded an excitatory amino acid transporter and the other a human homologue of the Drosophila disabled gene. This approach to the molecular biology of MS pathogenesis may help to illuminate previously unappreciated aspects of this disease. PMID- 9209266 TI - Kinin receptors on human neurones. AB - Knowledge of the distribution of kinin receptors in the human brain will aid our understanding of the role of kinins in neurophysiology. Furthermore, induction of the kinin B1 receptor may be important in the pathogenesis of neural diseases. Using polyclonal antibodies directed to specific regions of the B1 and B2 kinin receptors and standard immunolabelling techniques, we report on the localisation of these receptors on neurones in specific areas of the human brain. B2 bradykinin receptors are present in neurones of the brain stem, basal nuclei, cerebral cortex, thalamus and hypothalamus. B2 immunolabelling was also observed in the endothelial lining of the superior sagittal dural sinus and ependyma of the lateral and third ventricles. B1 kinin receptors have been localised on neurones of the thalamus, spinal cord and hypothalamus. Although binding of labelled bradykinin to neuronal membranes has been demonstrated, this is the first conclusive evidence for the existence of immunoreactive B1, and further confirmation of B2 receptors on human neurones. PMID- 9209267 TI - Astroglial overproduction of TGF-beta 1 enhances inflammatory central nervous system disease in transgenic mice . AB - Cerebral expression of the injury response cytokine transforming growth factor beta 1 (TGF-beta 1) has been found to be increased in several neurological diseases but it remains unclear whether its function is primarily beneficial or detrimental. Here we show that transgenic (tg) mice that overexpress bioactive (TGF-beta 1 in the central nervous system (CNS) and show no overt phenotype in the unmanipulated state, are more susceptible to the immune-mediated CNS disease experimental autoimmune encephalomyelitis (EAE). TGF-beta 1 tg mice with EAE showed an earlier onset of clinical symptoms, more severe disease and increased mononuclear cell infiltration in their spinal cords compared with non-tg littermate controls with EAE. Whereas previous observations indicated that increased peripheral levels of TGF-beta 1 can suppress EAE, our findings demonstrate that local expression of TGF-beta 1 within the CNS parenchyma can enhance immune cell infiltration and intensify the CNS impairment resulting from peripherally triggered autoimmune responses. PMID- 9209268 TI - Activation of nuclear factor-kappa B by beta-amyloid peptides and interferon gamma in murine microglia. AB - An increasing body of evidence suggests that amyloid-beta (A beta) peptides and microglia are crucially involved in the pathogenesis of Alzheimer's disease. In an effort to further elucidate the biological effects of A beta towards microglia, we investigated the ability of A beta peptides to activate nuclear factor (NF)-kappa B in the N9 murine microglial cell line. Co-stimulation of microglia with suboptimal concentrations of A beta(25-35) and 100 U/ml IFN gamma resulted in the detection of a specific NF-kappa B DNA-binding activity in nuclear extracts, as determined in gel mobility shift assays. This response required at least 120 min to be evident and supershift experiments revealed that the NF-kappa B complex contains both RelA and p50. Accordingly, immunoblot experiments showed that amongst NF-kappa B/Rel proteins, RelA and p50 are mobilized to the nucleus following microglial cell stimulation with A beta(25-35) plus IFN gamma. Higher concentrations of A beta(25-35) were effective by themselves in inducing NF-kappa B activation, both in the N9 microglial cell line and in rat primary microglia, as well as in human monocytes. For purposes of comparison, microglia were also stimulated with bacterial LPS, a known NF-kappa B inducer. As expected, LPS strongly induced the formation of two NF-kappa B DNA binding activities, one of which was identified as RelA/p50. The LPS response was also more rapid, as it was already evident by 40 min and remained sustained for up to 3 h. Collectively, these findings indicate that NF-kappa B activation might constitute one of the mechanisms underlying the inducible expression of kappa B dependent genes in microglia stimulated by A beta peptides and IFN gamma, or by LPS. PMID- 9209269 TI - Leukaemia inhibitory factor mRNA is expressed in the brains of patients with subacute sclerosing panencephalitis. AB - We have examined the in situ transcription of leukaemia inhibitory factor (LIF) in brain tissue from 3 cases of subacute sclerosing panencephalitis (SSPE) and in 2 non-neurological control brains. This has been compared with expression of interleukin 2 (IL-2), interleukin 6 (IL-6) and tumour necrosis factor beta (TNF beta) in the same tissues. All of the cytokines in the study were expressed in cells in the inflammatory infiltrate as well as in glial cells. LIF mRNA was also found to be expressed in neurons, in foci where these cells were also virally infected. No hybridization was found with any of the probes in areas of SSPE brain, which were negative for measles virus RNA or in the non-neurological control cases, although expression was demonstrated in the latter by use of reverse transcription-polymerase chain reaction (RT-PCR). Differentiated, cultured human neuronal cells were also positive, by RT-PCR, for LIF. This is the first demonstration of LIF expression in human brain and the results suggest that this cytokine is up-regulated, in several cell types, including neurons, following virus infection. PMID- 9209270 TI - Experimental immune-mediated damage of septal cholinergic neurons. AB - Degeneration of cholinergic neurons in the medial septum and the diagonal band of Broca is a frequent neuropathological feature of Alzheimer's disease. To determine whether an immune process can injure these basal forebrain cholinergic neurons, we serially immunized guinea pigs with septal cholinergic hybrid cells (SN-56). Following immunization, a relatively selective damage of septal cholinergic neurons, reduction in septal choline acetyltransferase (ChAT) activity and decrease in acetylcholine release in hippocampus were detected. Serum IgG from guinea pigs immunized with SN-56 cells and stereotactically injected into the medial septal region of rats produced a loss of ChAT activity in the medial septum, frontal cortex and hippocampus, together with impairment of learning and long term spatial memory. These data suggest that relatively selective damage to septal cholinergic neurons can be caused by an immune mediated process in experimental animals. PMID- 9209271 TI - The distribution and abundance of MHC and ICAM-1 on Schwann cells in vitro. AB - Schwann cells, the myelin forming glial cells of peripheral nerves, have been implicated as having an immunoregulatory role in inflammatory demyelinating neuropathies (IDNs) such as Guillain Barre syndrome (GBS) and chronic inflammatory demyelinating polyneuropathy (CIDP). We employed rat IFN-gamma, a cytokine released by macrophages and CD4+ T-cells during inflammatory demyelination of the peripheral nervous system, to examine the distribution and abundance of MHC class I, MHC class II and ICAM-1 on Lewis rat Schwann cells and fibroblasts in vitro. MHC class I, class II and ICAM-1 molecules were immunolabelled with 30 nm colloidal gold and observed by scanning electron microscopy. Incubation with IFN-gamma for 24 and 72 h, resulted in the clustering of MHC class I and ICAM-1 on Schwann cells and fibroblasts with MHC class II randomly distributed as single particles. MHC class I and ICAM-1 were upregulated after 24 h incubation in the presence of IFN-gamma, whereas MHC class II was upregulated after 72 h. The difference in the rate of upregulation may indicate differences in the recycling and/or synthesis of these molecules. Changes in distribution such as clustering, in conjunction with the upregulation of these molecules, suggest a role for Schwann cells in the restimulation of specifically primed CD4+ T-cells in IDNs. PMID- 9209272 TI - Expression of macrophage migration inhibitory factor in rat retina and its immunohistochemical localization. AB - Macrophage migration inhibitory factor (MIF) plays an important regulatory role for the T-cell activation induced by mitogenic or antigenic stimuli. We examined expression of MIF in rat retina. Reverse transcription-polymerase chain reaction analysis of a retinal tissue homogenate revealed that MIF mRNA was constitutively expressed. A single band specific for MIF protein was also observed by Western blot analysis. Immunohistochemistry using frozen sections of retinal tissues reacted with an anti-rat MIF antibody showed that MIF was localized in astrocytes. Muller cells and pigment epithelial (RPE) cells. This topological finding was confirmed by colocalization of MIF protein with glial fibrillary acidic protein and vimentin, which are putative immunohistochemical markers for astrocytes and Muller cells. It is well known that the retinal glial cells, as well as RPE cells, play an active role in inflammatory and immunological responses in the retina. Considering these facts, constitutive expression of MIF in these cells suggested the possibility that the protein contributes to regulation of retinal tissue inflammation as well as its local immunity. PMID- 9209273 TI - Anti-brain spectrin immunoreactivity in Alzheimer's disease: degradation of spectrin in an animal model of cholinergic degeneration. AB - In a previous work, we described the existence of anti-brain spectrin auto antibodies in Alzheimer's disease (AD) patients (J. Neuroimmunol. 68 (1996) 39 44). In this report, we further support our previous observations, showing that sera from 9 out of 18 AD patients, but none of 14 control subjects, immunoreacted with spectrin synthesized by PC12 cells. In addition, degradation of brain spectrin was found to be greatly enhanced in the frontal cortex of rats subjected to an animal model of cholinergic degeneration. Our data suggest that spectrin degradation and generation of anti-spectrin auto antibodies may be related to the cholinergic degeneration encountered in AD. PMID- 9209274 TI - Gender differences of inducible nitric oxide production in SJL/J mice with experimental autoimmune encephalomyelitis. AB - We identified gender related differences of inducible nitric oxide synthase (iNOS) expression and NO production in mice with experimental autoimmune encephalomyelitis (EAE). When myelin basic protein-specific T-lymphocytes derived from female mice were transferred, the female recipients developed more severe EAE and expressed higher levels of iNOS and NO than male recipients. When the T lymphocytes derived from males were transferred, severe EAE was induced in neither female or male recipients and neither iNOS nor NO were detectable. These data show an association between No production and EAE severity, suggesting a possible role of NO in the pathogenesis of EAE. PMID- 9209275 TI - Modulation of soluble and membrane-bound TNF-induced phenotypic and functional changes of human brain microvascular endothelial cells by recombinant TNF binding protein I. AB - In this study, the effects of TNF binding protein I (TBP I) on TNF-induced changes of human brain microvascular endothelial cells (MVEC) were investigated. TBP I completely abolished TNF-induced IL-6 production and E-selectin induction, while it partially inhibited TNF-induced IL-8 production and up-regulation of ICAM-1 and VCAM-1. Moreover, TBP I significantly inhibited TNF-induced cytotoxicity and leukocyte adherence on human brain MVEC. The inhibitory activity of TBP I for TNF was dose-dependent and related to the time of administration after TNF stimulation. In addition, TBP I inhibited membrane-bound TNF induced activation of human brain MVEC, but the concentration required was about 10-fold higher than that for soluble TNF. These results indicate a therapeutic potential for TBP I in diseases of the central nervous system associated with TNF overproduction. PMID- 9209276 TI - Induction of IP-10 chemokine promoter by measles virus: comparison with interferon-gamma shows the use of the same response element but with differential DNA-protein binding profiles. AB - Measles virus (MV) and interferon (IFN)-gamma induced IP-10 chemokine mRNA in U373 glioblastoma cells. The minimal response element for both MV and IFN-gamma was localized between nucleotide -231 and -153 of muIP-10 promoter, which contains an IFN-stimulated response element (ISRE) and the distal NF-kappa Bd site. Mutation of individual elements showed that ISRE and NF-kappa Bd were required to function together. DNA-protein binding profiles with the minimal response element showed that IFN-gamma induced a complex consisting of STAT1 while MV induced a complex consisting of p50 and p65 in the absence of new protein synthesis. IFN-gamma and MV also induced IRF-1 DNA binding activity which persisted for longer time periods with IFN-gamma stimulation. Despite the functional requirement of both ISRE and NF-kappa Bd elements, different combinations of DNA binding factors are used in the induction of IP-10 by MV or IFN-gamma. PMID- 9209277 TI - High levels of cerebrospinal fluid IgM binding to myelin basic protein are associated with early benign course in multiple sclerosis. AB - We assessed human myelin basic protein (MBP) binding IgM levels in CSF. MBP is the most studied putative antigen in multiple sclerosis (MS) and immune responses against it may be involved in the demyelination process. We also correlated these levels with EDSS score and other parameters of disease progression and prognosis, both at the time of CSF analysis and during follow-up. CSF IgM anti-MBP levels were assayed by measuring total IgM levels with solid-phase ELISA in CSF samples from 66 patients with relapsing-remitting MS, 11 subjects without neurological diseases, 20 patients with non-inflammatory neurological diseases and 7 patients with lymphocytic meningitis, before and after immunoabsorption with human MBP. Confirmation of IgM binding specificity was performed by immunoblotting of positive CSF samples onto MBP coated-nitrocellulose sheets. Clinical evaluation (disability score, number and time of attacks) was performed during a mean follow up of 2.7 +/- 1.1 years. 23 of the 66 relapsing-remitting MS patients (33.8%) had elevated IgM anti-MBP levels. In this patient subgroup, IgM anti-MBP levels correlated with the IgM index (r = 0.71; P = 0.0001), but not with CSF/serum albumin (r = 0.08; P = 0.72). In the first year of follow-up, patients with low IgM anti-MBP suffered from more numerous attacks than those with elevated levels (0.86 +/- 0.63 versus 0.43 +/- 0.58; P = 0.017). Patients with high IgM binding to MBP had a first attack during follow-up in a significantly higher time than those with low binding (28.87 +/- 4.7 versus 17 +/- 2.6 months, respectively; P = 0.005) and reached a decrease of 0.5 EDSS point significantly faster than those with low IgM (16.17 +/- 1.2 versus 29.7 +/- 2.6 months, respectively; P = 0.0002). A similar significant finding was observed when the time to reach low disability score (EDSS < or = 2.0) was analyzed (10.7 +/- 2.57 +/- 3.3 months, respectively; P = 0.014). These findings demonstrate that in a subgroup of MS patients, elevated CSF levels of IgM anti-MBP are associated with early favorable course and therefore suggest that IgM binding to MBP could be a possible prognostic marker in relapsing-remitting MS to select early MS patients for future trials. PMID- 9209278 TI - Howard Temin Memorial Lectureship. Molecular biology of HTLV-1: deregulation of host cell gene expression and cell cycle. PMID- 9209279 TI - Transcriptional activation and self-association in yeast: protein-protein dimerization as a pleiotropic mechanism of HTLV-I Tax function. AB - The yeast one-hybrid and two-hybrid systems for the detection of protein-DNA and protein-protein interactions were used as an in vivo approach to investigate the functional characteristics of HTLV-1 Tax expressed in yeast. Tax, when targeted to the upstream activating sequence (UAS) via the DNA-binding domain of Gal4 (Gal4BD), was found to activate a minimal promoter in yeast, indicating the presence of a functionally intact activation domain. Using the two-hybrid assay in which Tax was fused to either Gal4BD or Gal4 activation domain (Gal4AD), we demonstrate that Tax self-associates in the nucleus of yeast cells. Mutational analysis was performed to delineate the functional domain(s) necessary for Tax mediated trans-activation and self-association. Based on our results, we propose a pleiotropic mechanism in which Tax facilitates protein-protein dimerization of various cellular partners. PMID- 9209280 TI - HTLV-I Tax trans-activation and cell growth signaling. AB - We have cloned two genes for cell surface molecules, capable of delivering the intracellular signals, which are modulated for their expression by Tax. One is the gamma chain of the interleukin-2 (IL-2) receptor which is suggested to be critical for IL-2-dependent growth of human T-cell leukemia virus type I (HTLV-I) infected cells. The gamma chain is upregulated by Tax, like the IL-2 receptor alpha chain. This upregulation may compensate the gamma chain downregulation after IL-2 binding, presumably resulting in more frequent growth of HTLV-I infected T cells. The other is gp34 that was initially identified as a molecule specifically expressed on HTLV-I-infected T cells. gp34 has been demonstrated to bind OX40 which belongs to the tumor necrosis factor (TNF) receptor family. We found that HTLV-I Tax induces expression of gp34 and OX40, and that normal T cell transiently express both gp34 and OX40 upon antigenic stimulation. Collectively, it may be possible that HTLV-I-infected T cells are in a predisposition to growth due to modulated expression by HTLV-I Tax of gp34/OX40 and the gamma chain. PMID- 9209281 TI - Sp family members preferentially interact with the promoter proximal repeat within the HTLV-I enhancer. AB - Human T cell lymphotropic virus type I (HTLV-I) encodes the transactivator protein, Tax, which facilitates viral transcription from three 21 bp repeated elements within the U3 region of the long terminal repeat (LTR). Examination of the basal factors interacting with the 21 bp repeat elements through electrophoretic mobility shift (EMS) analyses has demonstrated the formation of DNA-protein complexes common to each of the 21 bp repeats (C1-C3) as well as three DNA-protein complexes specific to the promoter proximal (pp) repeat (U1 (U1A/U1B) and U2; 1-4). These studies have indicated that the individual repeats are not identical with respect to the cellular factors with which they interact. EMS analyses utilizing a series of mutated pp repeat elements demonstrate that the nucleotide sequence requirements for U1 (U1A/U1B) and U2 formation are separable from those required for C1-C3 formation. Competition EMS analyses utilizing Sp1 and CREB binding site oligonucleotides demonstrate that Sp family members are critical components of U1 (U1A/U1B) and U2 and that ATF/CREB family members are critical components of C1-C3. EMS supershift analyses have demonstrated that Sp1 is involved in U1A formation while Sp3 is involved in U1B and U2 formation. EMS analyses performed with nuclear extracts from Tax expressing Jurkat cells and HTLV-I-transformed peripheral blood mononuclear cells demonstrate that Tax prevents the formation of U1 (U1A/U1B) and U2 DNA-protein complexes. Therefore, Tax appears to inhibit the interaction of Sp family members with the pp repeat. Based on these observations, it is possible that the interaction of Sp and ATF/CREB family members with the pp repeat during basal and Tax-mediated transcription may play a critical role in viral gene expression during the initial stages of virus infection or during activation of a latent infection. PMID- 9209282 TI - HTLV-1 Tax protein interacts with cyclin-dependent kinase inhibitor p16Ink4a and counteracts its inhibitory activity to CDK4. AB - Tax, a regulatory protein of HTLV-1, is an oncoprotein which immortalizes human T cells and induces tumors in transgenic mice. Here, we found that Tax binds to a cyclin-dependent kinase inhibitor, p16Ink4a. p16Ink4a binds to cyclin-dependent kinases, CDK4 and CDK6, and inhibits their activity, resulting in suppression of G1 phase progression. The binding of Tax to p16Ink4a induced a reduction of p16Ink4a/CDK4 complex, with subsequent activation of CDK4 kinase. Tax also suppressed p16Ink4a-mediated inhibition of cell growth. The p16Ink4a gene was frequently deleted in many T-cell lines, but not in HTLV-1-infected T-cell lines. Taking these findings together, the functional inactivation of p16Ink4a by Tax through protein-protein interaction is suggested to contribute to cellular immortalization and transformation by HTLV-1. PMID- 9209283 TI - Antigenic variability of an immunodominant region (239-261) of the surface glycoprotein of HTLV-I. AB - In the 239-261 region of the surface glycoprotein of HTLV-I, we delineated five epitopes recognized by antibodies present in sera from HTLV-I infected patients. Three epitopes are located between the amino acids 252 and 261, one is of a linear type and the two others of a conformational type. One epitope is comprised between amino acids 244 and 249. The last epitope described lies between the amino acids 244 and 257 and strictly requires the presence of a serine at position 250 for its recognition. When patients are infected by viral isolates presenting a substitution of the serine at position 250 by a proline, no antibodies recognizing the 244-257 region could be found. Altogether, our data demonstrate that the immunodominant 239-261 region is complex and is subject to antigenic variability. PMID- 9209284 TI - AP-1 derived from mature monocytes and astrocytes preferentially interacts with the HTLV-I promoter central 21 bp repeat. AB - Characterization of the cellular transcription factors interacting with the human T cell lymphotropic virus type I (HTLV-I) long terminal repeat (LTR) is essential to dissecting the mechanisms involved in viral transcription that may be pertinent to the oncogenic and neuropathogenic processes associated with HTLV-I infection in both the immune and nervous systems. Electrophoretic mobility shift (EMS) analyses utilizing oligonucleotides homologous to each of the 21 bp repeat elements reacted with nuclear extracts derived from cell lines of lymphocytic, monocytic, neuronal, and glial cell origin have demonstrated differential binding of cellular factors to the three 21 bp repeats (1-4). ATF/CREB and Sp family members interacted with the 21 bp repeats to form DNA-protein complexes common to all cell types examined. However, a unique DNA-protein complex was detected when the promoter central 21 bp repeat was reacted with nuclear extracts derived from either the U-373 MG glioblastoma cell line or the THP-1 mature monocytic cell line. Based on nucleotide sequence requirements and immunoreactivity, we demonstrate that this DNA-protein complex is comprised of the AP-1 components, Fos and Jun. PMID- 9209285 TI - Expression of cell cycle-regulatory genes in HTLV-I-transformed T-cells and Tax1 immortalized T-cells. PMID- 9209286 TI - The role of topoisomerase I in HIV-1 replication. AB - The mechanisms involved in the restriction of the cellular tropism of HIV-1 to cells of primate origin remain to be clearly defined. However, a number of studies have shown that this is determined not only at the level of the cellular receptor(s) or virus entry, but at a number of additional and later stages in virus replication. We have recently reported that the reverse transcription of HIV-1 RNA is markedly enhanced by the association of the gag encoded nucleocapsid p15 protein and cellular topoisomerase 1. In the present study we have now investigated if the recruitment of cellular topoisomerase I during virus replication is important in determining the cellular tropism of HIV-1. Employing a stable murine cell line, L929, expressing both human CD4 and topoisomerase I, it could be demonstrated that effective proviral DNA synthesis occurred following infection. In contrast in cells expressing only human CD4 proviral DNA synthesis was not detected. In addition we have co-expressed fusin, a protein known to act as an accessory factor as the virus entry stage in infection of T cell tropic HIV 1, to support viral entry completely. However no progeny virus could be detected after HIV-1 infection. These results suggest that reverse transcription in vivo is critically dependent on the presence of cellular topoisomerase I, and support the view that involvement of this enzyme is in HIV-1 replication. Moreover the findings suggest that other factors which remained to be identified, are involved in restricting HIV-1 replication in non-primate cells. PMID- 9209288 TI - Chromosome changes associated with growth potential of HTLV-I infected human lymphocytes. AB - The association between chromosomal changes and cellular growth potential was investigated in HTLV-I infected human lymphocytes. Cell lines studied include Coculture-5 (HTLV-I infected immortalized cells dependent of IL-2), Coculture-15 and -18 (semi-transformed Coculture-5 cells following cocultivation with transformed Coculture-5 cells), UV-5 (Coculture-5 cells transformed following UV irradiation), and MNNG-1 (Coculture-5 cells transformed following MNNG treatment). Immortalized cells were grown in IL-2 medium (IL-2+PHA+TPA), whereas semi-transformed and transformed cells were grown in RPMI medium. By G-band karyotyping, double band formation was seen in 3-33% of spreads at the centromeric region of chromosomes 1 and 7 to which structural abnormalities were found to cluster. The double band formation was also seen by FISH using alpha satellite DNA probe in Coculture-5 and -15 thus far examined. The ploidy of immortalized, semi-transformed, and transformed cell lines, was 4n, 4n to 2n, and 2n range, respectively. These findings suggest that chromosomal rearrangements cause abnormalities in cell division kinetics resulting in numerical and structural abnormalities of chromosomes, and render host cells growth advantage. PMID- 9209289 TI - Cytokine mediated growth inhibition of HTLV-I infected transformed human lymphocytes. AB - The interaction between HTLV-I infected immortalized cells and transformed cells was studied. HTLV-I infected immortalized human lymphocytes (Coculture-5) dependent of IL-2 medium (IL-2+PHA+TPA) and transformed cells (UV-5) derived from Coculture-5 following UV irradiation were cocultured in equal numbers in IL-2 medium. UV-5 cells were suppressed and disappeared about 1 month in cocultivation. UV-5 cells were not suppressed in IL-2 medium. Coculture-5 cells and UV-5 cells were seeded in Transwell dishes that were separated by membrane with pores 0.4 micron in diameter and cultured in IL-2 medium. UV-5 cells were suppressed indicating that cell-cell contact was not required for the observed suppression. UV-5 cells were markedly suppressed in IL-2 medium harvested from Coculture-5 cells when recombinant interferon alpha (1000 U/ml) was added to the medium. These findings demonstrated that growth of HTLV-I infected transformed cells could be suppressed by cytokine(s) released by HTLV-I carrier lymphocytes. PMID- 9209287 TI - HIRF: a novel nuclear factor that binds to the human T-cell leukemia virus type I internal regulatory element (HIRE). AB - The transcription of human T-cell leukemia virus type I (HTLV-I) provirus starts from a promoter located in the 5' long terminal repeat (LTR). We have identified a second promoter at the 3' end of the pol gene. This internal promoter expresses the Tax transactivator protein, but does not require Tax for its activity. Furthermore, we have found the novel enhancer motif AGTTCTGCCC, which are located near the initiation site. We have named the sequence HIRE (HTLV-I internal regulatory element). The HIRE binding protein is a ubiquitous protein. We purified this protein from the HTLV-I producing cell line MT-2 cells by DNA affinity chromatography. SDS-PAGE analysis revealed four major bands (70, 85, 115 and more than 200 kDa) and some minor bands on the gel. We renatured each major protein and showed the 70 and 115 kDa proteins bind to DNA, although the 115 kDa protein seemed to bind nonspecifically. We have designated these components as HIRF (HTLV-I internal regulatory factor). PMID- 9209290 TI - Antibodies directed against a variable and neutralizable region of the HTLV-I envelope surface glycoprotein. AB - The majority of neutralizing antibodies of HTLV-I are directed against linear epitopes of the envelope surface glycoprotein (gp46) in the immunodominant region 175-199. Although gp46 presents a remarkable degree of conservation, the substitution of the proline at position 192 by a serine is described for 10 isolates among the 54 sequenced ones. This amino acid substitution is known to induce an important change in the orientation of the exposed residues of this region and has drastic consequences on the immunogenicity of the neutralizable epitopes located in this region. We developed monoclonal antibodies directed against epitopes located in this region containing a proline or a serine at position 192. The six monoclonal antibodies obtained recognize the gp46 at the surface of living HTLV-I producing cells, two of them are specific of a 190-197 epitope with a serine at position 192. This demonstrates that the antigenicity of this epitope differs depending on the presence of a proline or a serine at position 192. Altogether, these results demonstrate that the immunodominant neutralizable region 175-199 is antigenically variable. PMID- 9209291 TI - Inhibition of syncytium formation by antisense oligonucleotide phosphorothioates complementary to tax mRNA of human T-cell leukemia virus type 1 (HTLV-1). AB - HTLV-1 infection is known as the factor to cause adult T-cell leukemia (ATL). Antisense oligonucleotide phosphorothioates against tax gene and control oligonucleotide phosphorothioates were synthesized. Antisense oligonucleotide was complementary to the region of initiation codon of tax gene. Two control oligonucleotides were tax sense and random. HTLV-1-positive human T-cell line, C91/PL and HTLV-1 non-infected human glioma cell line, U251-MG were co-cultured in the presence of antisense or control oligonucleotides for 24 hours. Oligonucleotides used in this study were not toxic at 10 microM concentration. Antisense oligonucleotide against tax gene inhibited 59% the syncytium formation assay at 10 microM concentration. PMID- 9209292 TI - Substrates and inhibitors of human T-cell leukemia virus type 1 (HTLV-1) proteinase. AB - Human T-cell leukemia virus type 1 (HTLV-1) proteinase, a 125 residue polypeptide, was chemically synthesized using the solid phase method. The crude product was purified, renaturated and proteolytic activity was tested using oligopeptide substrates derived from processing sites of various retroviral polyproteins. Cleavage of the oligopeptide substrates together with an initial study using a series of HIV-1 and MAV (myeloblastosis associated virus) proteinase inhibitors suggest that the substrate specificity of HTLV-1 proteinase is very close to that of BLV (bovine leukemia virus) proteinase and distinct from that of both HIV-1 and MAV proteinases. PMID- 9209293 TI - Different properties of human T-cell leukemia virus type I prepared from 8C cat cells and human cells. PMID- 9209295 TI - Substitutions T-->C 4783 and T-->C 6569 in the pol and env genes of HTLV-I are characteristic for the isolates originating in the Middle East. PMID- 9209294 TI - Phylogenetic relationships of HTLV-I/STLV-I in the world. AB - In an effort to delineate the origin and evolution of HTLV-I/STLV-I, we have been conducting phylogenetic analyses on LTR sequences of this virus group. HTLV-I isolates newly analyzed in the present study were from Iran, South Africa, Cameroon, Sakhalin and Brazil where little is known concerning the genetic features of HTLV-I. In addition, STLV-I isolates were obtained from non-human primates in Africa and Asia including an isolate from orangutans in Indonesia. Proviral LTR sequences were amplified by nested PCR, and then sequenced. Phylogenetic trees were constructed by the neighbor joining method. The results obtained are: 1) African STLV-I isolates formed one large cluster together with the Central African group of HTLV-I in the tree; 2) Asian STLV-I isolates including that of an orangutan in Indonesia were highly divergent from African STLV-I and the Cosmopolitan group of HTLV-I, but not so closely related to each other and to the Melanesian group of HTLV-I; 3) An HTLV-I isolate of Cameroon Pygmy was related to African STLV-I isolates, but distinct from the Central African group of HTLV-I; 4) The majority of HTLV-I isolates belonged to subgroup A which is the most widespread subgroup of the Cosmopolitan group of HTLV-I, while some Brazilian isolates from descendants of Japanese immigrants belonged to subgroup B which mainly consists of HTLV-I isolates from Japan. 5) In the phylogenetic tree, several HTLV-I isolates of subgroup A from the same areas appear to form monophyletic clusters such as a subcluster of Brazilian and Colombian isolates and that of Iranian isolates. PMID- 9209296 TI - Kenneth MacGredie Memorial Lectureship. Adult T-cell leukemia/lymphoma. AB - Adult T-cell leukemia (ATL) was first reported in Japan, where it has a high incidence in the southwestern region. The retrovirus, human T-lymphotropic virus type I (HTLV-I), is the causative agent of ATL. In ATL-endemic areas, the rate of HTLV-I carriers is high. A definite diagnosis of ATL is based on the presence of HTLV-I proviral DNA in the tumor cell DNA. ATL cells originate from the CD4 subset of peripheral T cells. ATL shows diverse clinical features but can be divided into four subtypes: acute, chronic, smoldering, and lymphoma type. Chemotherapy is not effective; the acute and lymphoma types have a poor prognosis. Familial occurrence of ATL is common. HTLV-I infection is caused by transmission of live infected lymphocytes from mother to child, from man to female, or by blood transfusion. Infection with HTLV-I can lead to other diseases, including HTLV-I-associated myelopathy/tropical spastic paraparesis and HTLV-I uveitis. PMID- 9209297 TI - Primary prevention of HTLV-1 in Japan. AB - The Nagasaki Prefecture, Japan (population: 1.5 million), is one of the hot endemic foci of Human T-lymphotropic virus type 1 (HTLV-1). Prevalence of HTLV-1 carriers are approximately 10% in the age group over 40 years old (40,000 individuals), approximately 10 times of the national average. Annual registry of adult T-cell leukemia (ATL) in the Prefecture is approximately 60 cases (estimated incidence: 100 cases), or a half percent of total deaths. A effective measure to control the endemic cycle of HTLV-1 has been imperative, since practical ways to prevent or control ATL are not available. A prefecture wide intervention at Nagasaki by refrain from breast-feeding blocked approximately 80% of mother-to-child transmission of HTLV-1. PMID- 9209298 TI - Short-term breast-feeding may reduce the risk of vertical transmission of HTLV-I. The Tsushima ATL Study Group. AB - To establish a desirable preventive measure against mother-to-child transmission of HTLV-I through breast milk, we conducted a prospective study to investigate the seroconversion rate among children born to HTLV-I carrier mothers on two highly HTLV-I-endemic islands where 8% of pregnant women carry HTLV-I. Between 1985 to 1991, 428 pregnant women were found to be positive against anti-HTLV-I antibody and were advised not to breast-feed their newborn babies. Among them, 212 women (50%) accepted this advice and the other mothers proceeded to breast feed. Results were obtained from 277 children born to HTLV-I carrier mothers and were followed up until more than 30 months of age. When the seroconversion rate was analyzed by feeding manner, short-term breast-feeders (< or = 6 months) showed a statistically significant lower seroconversion rate than long-term breast-feeders (2/51; 3.9% vs. 13/64; 20.3%, p < 0.05). On the other hand, four out of 162 bottle-fed children (2.5%) became positive. It is hypothesized that maternal HTLV-I antibody may protect babies from HTLV-I infection through breast milk during the first 6 months. PMID- 9209299 TI - HTLV-I associated myelopathy (HAM): review and recent studies. PMID- 9209301 TI - HTLV-I provirus in the clinical subtypes of ATL. AB - Adult T cell leukemia (ATL) is an aggressive neoplasm of mature helper T cell, which is etiologically linked with human T-lymphotropic virus type 1 (HTLV-I). We studied HTLV-I provirus in 61 cases of ATL with Southern blot analyses and long PCR. These methods detected defective virus in 34 cases (56%). Furthermore, it found two types of defective virus. The first type (type 1) defective virus had both LTRs, but lacked internal sequences, such as gag and pol. Type 1 defective virus was seen in 50% of all defective virus. The second form (type 2) of defective virus had only one LTR, and 5'-LTR was preferentially deleted. This type of defective virus could be more frequently detected in aggressive types of ATL (16/44 cases) than chronic type (1/17 cases). This defective virus might be associated with clinical subtype. PMID- 9209300 TI - Pathogenesis and prevention of HTLV-1-associated diseases. AB - HTLV-1 is an important factor involved in various diseases including adult T-cell leukemia/lymphoma and HTLV-1 associated myelopathy/tropical spastic paraparesis. Amount of HTLV-1 provirus integrated in human peripheral blood mononuclear cells might be a candidate for a risk factor in the manifestation of HTLV-1 associated diseases. Experimental animal models would be useful to dissect the pathogenesis of HTLV-1 associated diseases. We present rat and mouse models of HTLV-1 infection. Using these animal models, we could clarify the intrauterine transmission of HTLV-1, and have found that both genetic background and HTLV-1 infection are important in pathogenesis of HAM/TSP-like rats. We also discuss the preventive measures of HTLV-1 transmission by use of antisense oligonucleotides. PMID- 9209302 TI - HTLV-I pX transgenic rats: development of cytokine-producing mammary carcinomas and establishment of the pX mammary carcinoma cell lines. AB - In two lines of transgenic rats (pX rats) from WKAH and F344 strains and carrying the HTLV-I pX gene under control of the mouse H-2Kd promoter, mammary carcinomas developed predominantly in females starting at about 5 months of age. The incidence of the tumor reached about 40% when the rats were 12 months old. Histology of the tumor was undifferentiated carcinoma with massive infiltration of granulocytes into the tumor tissue. Systemic granulocytosis and hepato splenomegaly due to extramedullary granulocytopoiesis were seen in pX rats and nude mice bearing pX mammary tumor. mRNAs of both pX and host genes, Gro and MIP 2, which are granulocyte chemoattractants of the IL-8 family, were highly expressed in the tumor tissue. Since expression and point mutation of several oncogenes and anti-oncogene, related with mammary carcinomas, were not demonstrated, hitherto unidentified novel oncogenic pathways may be transactivated by the pX transgene in these pX rats. pX mammary carcinoma cell lines, which have similar characteristics to the primary tumor, were established and the cells underwent apoptosis under the serum deprived conditions. The pX rats and the pX mammary carcinomas appear to be suitable models for analyses of HTLV-I pX oncogenesis and immune pathogenesis in vivo and in vitro. PMID- 9209303 TI - Immunopathogenesis of HTLV-I associated neurologic disease: massive latent HTLV-I infection in bone marrow of HAM/TSP patients. AB - The localization of mammalian retroviruses to specified immune organs has significant implications on the pathophysiology of retroviral associated diseases. Human T-cell Lymphotropic Virus Type I (HTLV-I) is considered a CD4+ lymphotropic virus although the virus has been shown to infect a large variety of cells in vitro. Similarly, the human immunodeficiency virus (HIV), once thought to be harbored only in CD4+ peripheral blood lymphocytes (PBL) has been shown to be present in latent form in lymph nodes of HIV infected patients. HTLV-I Associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP) is a chronic progressive neurologic disorder of the central nervous system and is believed to result from infection of HTLV-I in association with an immunopathogenic or autoimmune mechanism. Here we describe experiments which utilize the in situ hybridization/polymerase chain reaction technology to demonstrate extensive HTLV I infection of bone marrow in HAM/TSP patients. We discuss these results in the context of HTLV-I associated neurologic disease and extend these observations to other disorders of potential retroviral etiology and autoimmune involvement. PMID- 9209304 TI - Roles of JAK kinase in human GM-CSF receptor signals. AB - The IL-3 and GM-CSF (hGMR) receptors consist of two subunits, alpha and beta, both of which are members of the cytokine receptor superfamily. Phosphorylation of tyrosine residues of hGMR beta subunit and several cellular proteins are observed with hGM-CSF stimulation. We analyzed role of tyrosine residue of hGMR beta subunit and nature of tyrosine kinase, JAK2 in hGMR signals using several hGMR beta subunit mutants. In addition to box1 region, a membrane distal region (a.a. 544-589) of hGMR beta is required for c-fos activation. Only one tyrosine residue (Tyr577) exists within the region 544-589, and substitution of Tyr577 to phenylalanine in GMR beta 589 resulted in the loss of c-fos activation. In contrast, the same substitution in a wild type receptor did not affect GM-CSF induced activities such as c-fos mRNA induction and proliferation but abolished Shc phosphorylation. These results suggest that the activation of Shc is not essential for c-fos activation and several tyrosine residues co-ordinate to activate c-fos activation. It is well documented that IL-3 or GM-CSF activates JAK2 in BA/F3 cells. However the role of JAK2 in IL-3/GM-CSF functions is largely unknown. We examined the role of JAK2 in GM-CSF-induced signaling pathways. Dominant negative JAK2 (delta JAK2) lacking the C-terminus kinase domain, suppressed IL-3/GM-CSF induced c-fos activation, c-myc activation and proliferation suggesting that JAK2 is involved in both signaling pathways. PTP1D and Shc are phosphorylated by IL-3/GM-CSF in BA/F3 cells, however these phosphorylation events were inhibited by expression of delta JAK2. Taken together, these results indicate that JAK2 is a primary kinase regulating all the known activities of GM-CSF. JAK2 mediates GM-CSF induced c-fos activation through receptor phosphorylation and Shc/PTP1D activation. PMID- 9209305 TI - Pleiotropic expression of heterologous cytokine/receptor genes in HTLV-1 associated diseases: candidate TRS for chimeric gene therapy. AB - DNA motifs that encode for specific transcriptional regulatory sequences (TRS) when engineered adjacent to the structural protein coding domain of a suicide enzyme can provide cell-lineage specific protein expression. The disparate up regulation of several genes in adult T-cell leukemia (ATL) versus HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP), seropositive carriers (SPC) and uninfected normals may reflect events at the molecular level related to leukemogenesis or to processes maintaining the heme-oncologic phenotype. Further, the genetic transduction of cytokine and receptor genes uniquely associated with ATL may provide targets for the development of leukemia specific gene therapies aimed at exploiting differences in the production of certain growth factors and growth factor receptors. Comparisons of the transcriptional and translational levels of interleukin-2 receptor alpha chain (IL-2R alpha), transforming growth factor-beta 1 (TGF-beta 1) and intracellular adhesion molecule-1 (ICAM-1) in ATL, HAM/TSP, and SPC and in several control populations revealed selectively up-regulated expression in ATL. We evaluated the feasibility of using lymphoid-specific TRS to activate herpes simplex virus thymidine kinase (HSVtk) to achieve selective cytotoxicity in leukemias expressing terminal deoxynucleotidyl transferase (TdT). Selective and efficient leukemic cell killing was produced and suggests that similar chimeric gene constructs containing TRS elements for IL-2R alpha, TGF-beta 1, or ICAM-1 may prove useful in designing gene therapies to treat ATL. PMID- 9209306 TI - Neuropathology of HTLV-1-associated myelopathy (HAM/TSP). AB - In order to clarify pathogenesis of HAM/TSP, we performed a detailed neuropathologic analysis of seven autopsy patients with HAM/TSP. Inflammatory infiltrates of mononuclear cells and degeneration of myelin and axons were noted in the middle to lower thoracic spinal cords and were continuously extended to the entire spinal cord. Horizontal distribution of inflammatory lesions was symmetric at any spinal levels. Immunohistochemical analysis demonstrated T-cell dominance. The numbers of CD4+ T cells and CD8+ T cells were equally present in patients with shorter clinical course. Apoptosis of helper/inducer T cells were observed in the presence of TIA1+ cytotoxic T cells in these active inflammatory lesions. Inflammatory infiltrates were markedly decreased and CD8+/TIA1- T cells were predominated over CD4+ cells in patients with prolonged clinical course. HTLV-1 proviral DNA amounts in the freshly frozen spinal cord measured by quantitative PCR were well correlated with the numbers of infiltrated CD4+ cells. In situ PCR of HTLV-1 proviral DNA using multi-primary pairs demonstrated the presence of HTLV-1 infected cells exclusively in the mononuclear infiltrates of perivascular areas. From these findings, it is suggested that the target of the inflammatory process seen in HAM/TSP lesions may be HTLV-1 infected CD4+ T cells infiltrating the spinal cord. PMID- 9209308 TI - Cross-resistance analysis and molecular modeling of nonnucleoside reverse transcriptase inhibitors targeting drug-resistance mutations in the reverse transcriptase of human immunodeficiency virus. AB - Oxathlin carboxanilide analogs (UC) and alpha APA, compounds recognized as nonnucleoside reverse transcriptase (RT) inhibitors (NNRTI), were evaluated for activity against the human immunodeficiency virus (HIV-1) and drug-resistant variants. These NNRTIs are structurally diverse but potent inhibitors of HIV-1 with efficacy in the nanomolar to low micromolar concentrations. They interact at a specific site in the pain domain of the p66 subunit of RT. Treatment of HIV-1 infected cell cultures with UC compounds resulted in the selection of drug resistant viruses bearing specific amino acid changes at 100, 101, 103, 106, and/or 181. Since Y181C and L1001 are the most commonly observed resistance engendering mutations, RT enzymatic analysis was correlated with molecular modeling to glean information on the structural interactions between these NNRTIs and RT. Information derived from these studies will facilitate rational drug design and the selection of complementary anti-HIV drugs for combination therapy. PMID- 9209307 TI - Increased fidelity of drug-selected M184V mutated HIV-1 reverse transcriptase as the basis for the effectiveness of 3TC in HIV clinical trials. AB - HIV-infected individuals, who received 3TC monotherapy over one year, generally had lower plasma viral burden than at base-line. This was in spite of high-level resistance to this compound and the appearance of the M184V substitution in the HIV reverse transcriptase (RT) gene, responsible for diminished sensitivity to 3TC. This apparent contradiction is explained by an increase in the fidelity of the HIV RT, conferred by the M184V mutation, on the basis of the following observations. First, titers of viral neutralizing antibodies, as measured against sequential autologous HIV isolates, remained stable in this population in contrast to rapid declines in patients treated with other drugs. This suggests that increased fidelity of M184V RT may limit variability in the HIV env gene and result in protracted effectiveness of anti-viral immune responsiveness. Second, recombinant HIV, that contained the M184V substitution in RT, could not replicate in the presence of d4T, AZT, Nevirapine, Delavirdine or Saquinavir, using previously described protocols for the generation of drug resistance in vitro. PMID- 9209309 TI - Genetic variation within human immunodeficiency viruses generates rapid changes in tropism, virulence, and transmission. AB - The human immunodeficiency viruses (HIV-1) undergo high rates of variation. Only a few point mutations in the envelope gene are required to switch the tropism of HIV-1 from a growth preference for monocytes to lymphocytes or to acquire lytic properties for rapid killing of infected T4 lymphocytes. Since heterosexual transmission efficiency is high for HIV-1's that are most prevalent in Africa or Asia, but low for HIV-1 B, which dominates in the US and western Europe, we asked whether African and Asian viruses had a particular tropism for cells of the reproductive tract. Langerhans' cells (LC), showed only minimal susceptibility to infection with HIV-1B from the US, but substantially greater sensitivity for infections by HIV-1 E and HIV-1 C, subtypes that predominate in Asia and Africa. PMID- 9209310 TI - SIV/HIV-1 chimeric viruses having HIV-1 env gene: a new animal model and a candidate for attenuated live vaccine. AB - In order to generate an HIV-1, which is infectious to and induces AIDS-like disease in monkeys, an SIVmac/HIV-1 chimeric virus, designated NM-3rN, which was replication-competent in monkeys, was constructed by recombination between HIV-1 and SIV mac genomes. The NM-3rN enabled to evaluate the efficacy of HIV-1 Env directed vaccines using macaque monkeys instead of chimpanzees, because NM-3rN had HIV-1 derived Env. Other NM-3rN-derivative chimeric viruses, designated NM-3 and NM-3n, which had defective vpr (plus nef for NM-3) genes, induced long-term persistent infection in monkeys having long-lasting humoral and cell-mediated immune reactions without manifesting the disease. The challenge inoculation with NM-3rN to these defective chimeric virus-infected monkeys resulted in protection. Furthermore, these protected monkeys were also resistant to challenge with another chimeric virus having different antigenicity in V3 loop from that of NM-3 and NM-3n. These results indicate that SIV/HIV-1 chimeric viruses may be a potential candidate for development of anti-AIDS live attenuated vaccines. PMID- 9209311 TI - Mechanism(s) of FIV vaccine protection. AB - Infection of domestic cats with the feline immunodeficiency virus (FIV) represents an important veterinary health problem and a useful animal model for the development of vaccines against AIDS. Two experimental FIV vaccines have been developed: one consisting of fixed infected cells and the other of inactivated whole virus. Both vaccines elicited strong CTL responses to FIV and high virus neutralizing (VN) antibody titers. Over 90% of vaccinated cats were protected against intraperitoneal infection with 10 cat infectious dose 50% (10 CID50) of either homologous FIVPet or heterologous FIVDix. As a means to evaluate the mechanism of vaccine protection, cats were either passively immunized with serum antibodies or transfused with peripheral blood cells from FIV-vaccinated cats. Cats passively immunized with vaccine sera or purified vaccine antibodies were protected from homologous FIV infection at a challenge doses which infected all control cats (5 and 10 CID50). Such passive protection was not achieved against heterologous FIV challenge. More importantly, cats transfused with washed blood cells from half-matched vaccinated cat were protected from FIV challenge (20 and 50 CID50). As expected, protection was not observed in cats transfused with cells from either unmatched vaccinated or half-matched unvaccinated cats. Further, only peripheral-blood cells from vaccinated cats had FIV-specific CTL responses to both autologous and half-matched target cells. Overall, these findings suggest that both humoral and cellular immunity are required for optimal vaccine protection. PMID- 9209312 TI - Mutational analysis of the 5' noncoding region of human immunodeficiency virus type 1 genome. AB - Retrovirus particles are released by budding from the membranes of infected cells. In the course of virus production, particularly during the late stage, viral genomic RNA is incorporated specifically into virion particles. This specific incorporation of the genomic RNA requires a packaging signal sequence. A region that functions as the packaging signal was mapped to a location upstream of the gag open reading frame on the HIV-1 viral genome. In addition of this packaging signal, other cis-acting elements that are scattered throughout the genome are also required for efficient packaging. The region upstream of the splice donor site is probably important for dimer formation. Therefore, we focused on one region located between the 3' end of the primer binding site and the 5' splice donor site of HIV-1. Experiments were conducted to investigate how deletions or point mutations in this region affect both dimerization in vitro and the production of infectious virus particles. A series of RNAs of varying lengths containing the 5' noncoding region were generated, and genomic dimerization of the altered viral RNA was analyzed in vitro. One RNA construct which consisted of 112 nucleotides (nt) from nt 639 to nt 750 formed a heterodimeric complex with the RNA which consisted of 200 nucleotides from nt 551 to nt 750. We then constructed proviruses with mutations in the 639 to 750 nt region and assayed for virus production. Several mutants that lacked the complementarity necessary to form a possible stem-loop structure in this region showed decreased production of infectious virus particles. Moreover, both deletion of this region and randomization of its nucleotide sequence completely impaired infectious virus production. Thus, the way that this region affects infectious virus production may be through its RNA secondary structure. PMID- 9209313 TI - Suppression of retrovirial replication: inactivation of murine leukemia virus by compounds reacting with the zinc finger in the viral nucleocapsid protein. AB - All retroviral nucleocapsid (NC) proteins, except those of spumaretroviruses, contain one or two zinc fingers, consisting of the sequence C-X2-C-X4-H-X4-C. Rice et al. (Science 270:1194-1197, 1995) have described a series of compounds which inactivate HIV-1 particles and oxidize the sulfur atoms in the NC zinc finger. We have characterized the effects of three such compounds on Moloney murine leukemia virus (MuLV). We find that, as with HIV-1, the compounds inactivate cell-free MuLV particles and induce disulfide cross-linking of NC in these particles. In contrast, the compounds have no effect on the infectivity of human foamy virus, a spumaretrovirus lacking zinc fingers in its NC protein. The resistance of foamy virus supports the hypothesis that the zinc fingers are the targets for inactivation of MuLV and HIV-1 by the compounds. The absolute conservation of the zinc finger motif among oncoretroviruses and lentiviruses, and the lethality of all known mutations altering the zinc-binding residues, suggest that only the normal, wild-type structure can efficiently perform all of its functions. This possibility would make the zinc finger an ideal target for antiretroviral agents. PMID- 9209314 TI - High levels of viremia in hu-PBL-NOD-scid mice with HIV-1 infection. AB - We studied the compatibility of human lymphocyte engraftment and susceptibility to HIV-1 infection in 2 new immunodeficient mice. NOD/Shi-scid mice were generated by backcrossing of the scid mutation into NOD mice while C57BL/6 RAG2(0/0) were generated by knocking out the RAG-2 gene. Human T lymphocytes were reconstituted in new immunodeficient mouse strains. We found that the new immunodeficient mouse strains accepted human PBL engraftment and HIV-1 infection more efficiently than conventional C.B-17-scid mice. Especially in the hu-PBL NOD/Shi-scid strain, we reproduced the high levels of HIV-1 viremia comparable to or at significantly higher levels than after HIV-1 primary infection. These results indicate that our hu-PBL-NOD-scid animal is useful for investigations of the activation mechanism in HIV-1 replication in vivo and after primary infection. PMID- 9209315 TI - The role of topoisomerase I in HIV-1 replication. AB - The mechanisms involved in the restriction of the cellular tropism of HIV-1 to cells of primate origin remain to be clearly defined. However, a number of studies have shown that this is determined not only at the level of the cellular receptor(s) or virus entry, but at a number of additional and later stages in virus replication. We have recently reported that the reverse transcription of HIV-1 RNA is markedly enhanced by the association of the gag encoded nucleocapsid p15 protein and cellular topoisomerase I. In the present study we have investigated if the recruitment of cellular topoisomerase I during virus replication is important in determining the cellular tropism of HIV-1. Employing a stable murine cell line, L929, expressing both human CD4 and topoisomerase I, it could be demonstrated that effective proviral DNA synthesis occurred following infection. In contrast in cells expressing only human CD4, proviral DNA synthesis was not detected. In addition we have co-expressed fusin, a protein known to act as an accessory factor as the virus entry stage in infection of T cell tropic HIV 1, to support viral entry completely. However no progeny virus could be detected after HIV-1 infection. These results suggest that reverse transcription in vivo is critically dependent on the presence of cellular topoisomerase I, and support the view that involvement of this enzyme is important in HIV-1 replication. Moreover the findings suggest that other factors which remained to be identified, are involved in restricting HIV-1 replication in non-primate cells. PMID- 9209316 TI - Acquired cytotoxic activity of CD8+ HIV-1IIIB carrier immature T cell clones specific to CD4+CD8+ (parental) human T cell clones and normal thymocytes. AB - By infecting human leukemia cell lines in vitro with HIV-1IIIB' a number of HIV-1 carrier clones were generated. Among them, 5 of 13 CD8+ HIV-1 carrier T cell clones were shown to acquire a rapid cytotoxic activity (within 1 hour) specific to TdT+CD4+CD8+ immature T cells including normal thymocytes. This novel cytotoxic reaction, without requiring virus infection and indicating a rapid T cell precursor elimination during active lymphopoiesis, suggests a mechanism responsible for mature CD4+ T cell depletion in HIV-1 infected individuals. PMID- 9209317 TI - Peptide inhibitors of HIV-1 and HIV-2 proteases: a comparative study. AB - HIV-1 and HIV-2 proteases (PR) which play the key role in the formation of infectious viral particles offer a target for inhibitors that could block the maturation step. Inhibitors o HIV-1 PR exhibit mostly 1-2 orders of magnitude weaker affinity for HIV-2 PR. The subsite specificity study of the HIV-1 and HIV 2 proteases performed with inhibitors varying in the type of nonhydrolysable bonds and amino acid residues in the P1, P1'and P2'positions has led us to the design of inhibitors with 2S,4S and 2R,4S stereomeres of the hydroxyethylene isostere and Glu or Gln in the P2'positions. These compounds inhibit HIV-1 and HIV-2 proteases in vitro in subnanomolar concentrations and exhibit the activity in tissue culture. PMID- 9209318 TI - Lifetime treatment of mice with azidothymidine (AZT) produces myelodysplasia. AB - AZT has induced a macrocytic anemia in AIDS patients on long term AZT therapy. It is generally assumed that DNA elongation is stopped by the insertion of AZT into the chain in place of thymidine thus preventing the phosphate hydroxyl linkages and therefore suppresses hemopoietic progenitor cell proliferation in an early stage of differentiation. CBA/Ca male mice started on AZT 0.75 mg/ml H2O at 84 days of age and kept on it for 687 days when dosage reduced to 0.5 mg/ml H2O for a group, another group removed from AZT to see recovery, and third group remained on 0.75 mg. At 687 days mice that had been on 0.75 mg had average platelet counts of 2.5 x 10(6). Histological examination on 9 of 10 mice with such thrombocytopenia showed changes compatible with myelodysplastic syndrome (MDS). A variety of histological patterns was observed. There were two cases of hypocellular myelodysplasia, two cases of hypersegmented myelodysplastic granulocytosis, two cases of hypercellular marrow with abnormal megakaryocytes with bizarre nuclei, one case of megakaryocytic myelosis associated with a hyperplastic marrow, dysmyelopoiesis and a hypocellular marrow and two cases of myelodysplasia with dyserythropoiesis, hemosiderosis and a hypocellular marrow. Above mentioned AZT incorporation may have induced an ineffective hemopoiesis in the primitive hemopoietic progenitor cells, which is known to be seen commonly in the myelodysplastic syndrome. PMID- 9209319 TI - Anti-human immunodeficiency virus activity of oligosaccharides from rooibos tea (Aspalathus linearis) extracts in vitro. AB - The active substances, acid polysaccharides, were extracted with 1% sodium hydroxide from the leaves of rooibos tea (Aspalathus linearis), Du Zhong Cha (Eucommia ulmoides Oliv.) and Japanese tea leaves (Camellia sinensis var. sinensis). The alkaline extracts of Rooibos tea and Du-Zhong tea leaves, but not Japanese tea leaves suppressed the HIV-induced cytopathicity using HIV (HTLV-III) infected MT-4 cells, having extremely low cytotoxicity: Its 50% effective concentration (EC50) was 12-67 micrograms/mL, white 50% cytotoxic concentration (CC50) was higher than 1.0 mg/mL. The active substances were purified with ethanol precipitation. The substances were composed of 27% of reducing sugar, 46% of neutral sugars and 22% of uronic acid. A LD50 of the alkaline extracts from rooibos tea was higher than 1.2 g/kg body weight. Acid degradated substances composed of disaccharides and trisaccharides, were also suppressed the HIV induced cytopathicity. From these results, it is probable that acid polysaccharides from rooibos tea were extremely safe, and that HIV infection may be suppressed by daily intake of the alkaline extracts of rooibos tea and Du Zhong tea. PMID- 9209320 TI - In vivo sequence changes of HIV-1 envelope V2 and V3 loops. AB - Human immunodeficiency virus type 1 (HIV-1) circulates in vivo as heterogeneous mixed populations (quasispecies). We analyzed the quasispecies nature of the first, second, and third hypervariable loops (V1, V2, and V3) of the gp 120 in plasma and viral isolates obtained from 19 infected individuals. Sequence analysis of 7 SI and 10 NSI isolates showed that the increased positive charge in V3 and elongation of V1 clearly correlated with viral SI capability in PBMC. Contrary to the previous report (M. Groenink et al. Science 260: 1513-1525, 1993), there appeared to be no correlation between SI or non-SI phenotype and length of V2. However, all the 5 isolates with long V2 and basic V3 showed SI phenotype, whereas 4 out of 6 isolates with short V2 and basic V3 showed NSI phenotype, still suggesting some functional cooperation of V2 and V3 on viral syncytium inducing capability. Sequence analysis of HIV-1 in plasma showed the positive correlation between V3 charge and V1 or V2 length in 4 cases. This result suggests the associated evolution of V1/V2 and V3 in these cases. PMID- 9209321 TI - Inhibition of HIV-1 replication by targeting the Rev protein. AB - Human immunodeficiency virus type 1 (HIV-1) encodes two regulatory proteins, Tat and Rev. The Rev protein facilitates the transport of unspliced and singly spliced RNA to the cytoplasm in infected host cells by binding to target RNA (Rev response element: RRE). A variety of approaches targeting Rev function, including gene therapy, have been developed that inhibit HIV-1 replication in cells cultured in vitro. This minireview summarizes the recent developments as well as our application of the Rev-binding element-based decoy approach using RNA-DNA chimera oligonucleotide modeling. PMID- 9209323 TI - Tissue and differentiation specific expression on the endogenous retrovirus ERV3 (HERV-R) in normal human tissues and during induced monocytic differentiation in the U-937 cell line. AB - ERV3 (HERV-R) is a complete, single copy human endogenous retrovirus located on the long arm of chromosome 7. The open reading frame in its envelope gene has been conserved during evolution but the gag and pol genes contain in-frame termination codons. To find a suitable experimental model system for analysis of the functions of the ERV3 genome, an extensive screening study of different normal and neoplastic human tissues was performed. Most tissues express low levels of the ERV3 env mRNA although high expression levels are observed in placenta, sebaceous glands, adrenals, testis, bronchial, epithelium and the monocytic cell line U-937. In U-937 cells the ERV3 env expression varied in a manner related to the differentiation status of the cells; being highest in the terminally differentiated non proliferating cells. U-937 cells can be induced to differentiate from the monoblastic to the mature monocyte/macrophage stage upon stimulation by several substances such as phorbolesters (TPA), Vitamin D3, Retinoic Acid (RA) and combinations of some cytokines. We conclude that the ERV3 locus is expressed in a tissue and differentiation specific way and that the U 937 cell line is a suitable model system to further analyze the proposed functions of ERVs such as immunomodulation, cell fusion and protection against exogenous retroviral infections. PMID- 9209322 TI - Molecular analyses of HIV-1 group O and HIV-2 variants from Africa. AB - Genetic variation among HIV isolates creates challenges for their detection by serologic and genetic techniques. To characterize the sequence variation and its correlation to serologic diversity of HIV-1 Group O and HIV-2 isolates, samples were identified by differential reactivity in selected commercial and research assays. Analysis of sera from Equatorial Guinea (EG) led to identification of 4 HIV-1 Group O variants. Viral RNA, extracted from these samples was used to PCR amplify overlapping sequences of the entire envelope gene using multiple primer pairs. Sequence analysis indicated that the V3 loop nucleotide and protein sequences aligned more closely with HIVANT70 compared to other Group O sequences. The amino acid sequences at the octameric tip of the V3 loop were RIGPLAWY, RIGPMAWY, or GLGPLAVY. The tetrameric tip GPLA is represented only once in the published 1994 HIV database (Los Alamos) but was present in 2 of 4 of EG samples. The immuno-dominant region (IDR) sequences derived from EG sera were unique in that none of the sequences were completely homologous to other HIV-1 group O variants. Further, the HIV-1 group O sequence variation could be correlated with differential serologic reactivity using IDR peptides. Compared to HIV-1, the sequence information on HIV-2 isolates is relatively limited, though the HIV-2 isolates also show genetic variation similar to HIV-1. To further establish a correlation between the genetic diversity and serologic detection of HIV-2, plasma samples from Western Africa were evaluated. Eight samples were selected based on weak serologic reactivity to env proteins. PCR amplification and sequence analysis of the gag, env V3 loop, and env IDR regions indicated that the samples could be classified as subtypes A (4 samples), B (3 samples) and D (1 sample). Across the subtypes, there was conservation in the IDR region of the sequence WGCAFRQVCHT. This region is absolutely conserved among the majority of currently known HIV-2 and related SIV viruses (1994 HIV database). One subtype B sample had a unique sequence immediately adjacent to the IDR, however, this did not change the serologic detection using a HIV-2 IDR specific monoclonal antibody. PMID- 9209325 TI - Charlotte Friend Memorial Lecture: murine leukemia virus (MuLV) tumorigenesis. AB - Recent analysis of over 500 lymphomas occurring in NFS.V mice, congenic for Akv type ecotropic MuLV structural genes, has revealed that about 90% are of B cell lineage as determined by demonstration of clonal rearrangements of Ig heavy chain genes, phenotyping by immunocytochemistry or cytofluorometric analysis, and by site and morphology of tumor. At least 40% of the B cell lymphomas were found to have their origin in the splenic marginal zone, a site only once before described for mouse lymphomas. Clonal somatic integrations of ecotropic MuLV occurred in 85% of tumors. PMID- 9209324 TI - Human endogenous retroviruses: expression in various organs in vivo and its regulation in vitro. AB - To evaluate the role of human endogenous retroviruses in vivo, we examined their expressions in various organs from autopsy cases by Northern blot and RT-PCR. ERV3 (HERV-R) mRNA was expressed in many organs, and the level of expression in individuals and organs considerably differed. However, expression in the adrenal gland showed consistently high levels in every individual. lambda 4-1 (HERV-E) mRNA was expressed less compared with that of ERV3, and could not be detected in the adrenal gland by Northern blot, although the expression of lambda 4-1 generally correlated with that of ERV-3 in placentas. We also examined the effect of cytokines on the transcriptional regulation of ERV3 in vitro. Although the level of ERV3 expression in cultured synovial cells did not change after IL-1 beta treatment, the level in cultured proximal tubular epithelial cells was upregulated. The evidence suggests that distinct regulatory pathways may exist for the expression of human endogenous retroviruses in different cell types. PMID- 9209326 TI - Early (preleukemic) events in Moloney murine leukemia virus leukemogenesis. PMID- 9209327 TI - Pathogenesis of Friend leukemia virus. AB - Friend leukemia virus complex (FLV) consists of replication-defective, Friend spleen focus-forming virus (F-SFFV) and replication-competent, Friend murine leukemia virus (F-MuLV). We produced transgenic mice possessing F-SFFV gp55 gene and clarified that the gp55 glycoprotein encoded by F-SFFV env-related gene is, by itself, responsible for the initiation of erythroleukemia. The occurrence of erythroleukemia, however, is sporadic in these mice. Erythroleukemia cell lines established from these mice possessed mutations in the p53 allele. One had a temperature-sensitive mutant p53 allele, p53Val-135 and showed induction of apoptosis by expressing a wild-type p53 protein at 32 degrees C. Superinfection of the mice with Moloney murine leukemia virus (Mo-MuLV) conferred 100% induction of erythroleukemia, mutating p53 gene or activating Spfi-1 gene by insertional events. Activation of the JAK/STAT pathway, which is involved in cytokine signaling, was investigated in the gp55 signaling mediated by the erythropoietin receptor. JAK1 and STAT5 were constitutively tyrosine-phosphorylated but the DNA binding activity of STAT5 was not induced. PMID- 9209328 TI - Fre-2, a locus closely linked to Fv-2, is rearranged in some erythroleukemias induced by Friend murine leukemia virus. AB - Friend murine leukemia virus (F-MuLV) induces leukemia by integration into the cellular genome, thereby changing the structure of expression of cellular oncogenes. Here we describe a new F-MuLV integration site Fre-2 isolated from splenic DNA of an erythroleukemic animal. This site has been found rearranged in 5 out of 63 additional tumors; however, no F-MuLV proviruses could be detected in the vicinity of the rearrangement sites in these 5 cases. The rearrangements represented closely clustered chromosomal breakpoints, presumably chromosomal translocations. Exons transcribed into differentially spliced mRNAs of 1.9 and 3.7 kb have been found near the breakpoint. No sequences that are homologous to Fre-2 could be found in databases. PMID- 9209329 TI - Mutational analysis of the structure-function relationship of the leukemogenic membrane glycoprotein (GP55) of Friend spleen focus-forming virus (F-SFFV). AB - Friend spleen focus-forming virus (F-SFFV) causes acute erythroleukemia in adult mice. F-SFFV encodes an envelope protein-like membrane glycoprotein called gp55 in its defective env gene. Gp55 is responsible for the early stage of leukemogenesis by F-SFFV by specifically binding to and activating the murine erythropoietin receptor (EPO-R). Gp55 has a polytropic env sequence in its N terminal portion. This portion probably contains the binding site for the EPO-R. In order to obtain a clue for the structure of the binding site to the EPO-R, we isolated and analyzed many spontaneous revertant F-SFFVs which derived from the non-leukemogenic mutant F-SFFV having an ecotropic env sequence instead of the polytropic env sequence in its gp55 gene. PMID- 9209330 TI - Interaction of the MAIDS defective virus with its host. PMID- 9209331 TI - Possible origin of murine AIDS-inducing sequence. AB - The murine AIDS (MAIDS) virus has a unique sequence in its gag p12 region. A transcript which hybridizes with this sequence is expressed in normal C57BL/6 mice. This transcript has been proposed to be the origin of the MAIDS virus, since the virus was originally isolated from radiation-induced leukemic C57BL/6 mice. The transcript, designated Edv, was molecularly cloned and sequenced. Compared with the nucleotide sequence of the helper LP-BM5 ecotropic virus, the pathogenic defective MAIDS virus has 16-bp deletions and a 1-bp insertion in the 5' and 3' regions of the gag p12 sequence, respectively, and the Edv transcript contains only a 3-bp deletion. Therefore, the amino acid sequence of the gag p12 region of the MAIDS virus is less homologous to that of the helper virus and Edv transcript due to the frameshift mutations. This suggested that the MAIDS virus was generated by such frameshift mutations in the gag p12 region during recombination between the helper virus and the Edv or a related sequence. PMID- 9209332 TI - Immunologic havoc induced by the 60 kD Gag protein of the MAIDS defective virus. AB - The mechanisms responsible for development of profound immunodeficiency and extensive lymphoproliferation that characterize infection of different species with retroviruses are only partially understood. In mice, it has been shown the activities of an unusual Gag protein are necessary and sufficient to induce these abnormalities in a syndrome designated mouse AIDS (MAIDS). Current studies suggest that complex, antigen-driven interactions between T cells and B cells result in polyclonal activation of both types of lymphocytes, aberrant cytokine production and late lymphomas. PMID- 9209333 TI - Plasmacytoid leukemia of chinook salmon. AB - Plasmacytoid leukemia is a common disease of seawater pen-reared chinook salmon (Oncorhynchus tshawytscha) in British Columbia, Canada, but has also been detected in wild salmon, in freshwater-reared salmon in United States, and in salmon from netpens in Chile. The disease can be transmitted under laboratory conditions, and is associated with a retrovirus, the salmon leukemia virus. However, the proliferating plasmablasts are often infected with the microsporean Enterocytozoon salmonis, which may be an important co-factor in the disease. PMID- 9209334 TI - Molecular characterization of a piscine retrovirus, walleye dermal sarcoma virus. PMID- 9209335 TI - Avian leukemias and lymphomas: interplay between retroviruses and herpesviruses. AB - Avian leukemias and lymphomas are caused primarily by retroviruses and herpesviruses. The protooncogenes activated by avian retroviral insertions in B & T-cell lymphomas will be summarized, with discussion on a new common insertion site, bravo, associated with RAV-O LTR insertion. Two novel interactions between avian retroviruses and Marek's disease herpesvirus (MDV) will be described: one involves direct interactions between putative viral oncoproteins and the other integrative recombination between these two viruses. PMID- 9209336 TI - Rapid induction of B-cell lymphomas by avian leukosis virus. AB - Avian leukosis viruses (ALVs) that induce rapid B-cell lymphomas integrate into the c-myb gene and produce an ALV-myb read-through RNA, which is spliced to produce a truncated Myb protein. The genetic determinants of such recombinant ALVs have been mapped to a 42-nt deletion within the gag gene. This deletion increases splicing efficiency since it is located within a negative regulator of splicing. We propose that the deletion leads to increased production of Myb protein by increasing splicing of an ALV-myb pre-mRNA. PMID- 9209337 TI - Mouse mammary tumor virus: a virus that exploits the immune system. AB - Mouse mammary tumor virus (MMTV) causes mammary carcinomas and T-cell tumors in mice. MMTV variants that induce T-cell tumors have a large deletion within the U3 region of the long terminal repeat (LTR) compared to MMTV strains that induce mammary tumors. We provide evidence here that T-cell tropic MMTV strains lack a redundant binding site for a cellular protein called NBP (negative regulatory element binding protein). The lack of NBP-binding sites in T-cell tropic MMTV strains presumably leads to higher levels of transcription in T-cells during the MMTV life cycle and an increased incidence of mutagenic integration events. PMID- 9209338 TI - Possible relationships between canine hematopoietic neoplasia, other malignancies and immune mediated diseases. AB - In the last ten years few retroviruses have been isolated. The reasons for this lack of new discoveries are obvious, most retroviral investigations have been directed to HIV, HTLVI and HTLVII research with little research attention given to other species that may have retroviral infections, such as the dog. The domesticated dog is particularly important to investigate because they live in close proximity to human beings and have done so for at least the last 14 thousand years (1). This report gives relationships between canine hematopoietic, other neoplasia and immune mediated diseases in two families of inbred dogs that possibly were also infected with a yet to be fully described retrovirus. These findings should lead to a renewed interest in studying retroviral infections in dogs. The heterospecies relationship of retroviral infections closely related to human AIDS virus is of obvious scientific importance. PMID- 9209339 TI - Regulation of gene expression directed by the long terminal repeats of feline leukemia viruses. AB - Repetitive structure of enhancer elements in the long terminal repeat (LTR) has been identified in feline leukemia viruses (FeLVs) integrated in lymphoid tumor cells in cats. In this study, promoter activities of the FeLV LTRs were measured in lymphoid and non-lymphoid cell lines in transient expression assays using plasmids containing the viral LTRs linked to the chloramphenicol acetyltransferase (CAT) gene. Promoter activity of the LTR with 3 enhancer repeats (pFTLTR) was significantly higher than that of the LTR with 1 enhancer (Glasgow-1 LTR) in feline (FT-1) and human (Jurkat) T-lymphoblastoid cell lines. Promoter activity of the pFTLTR was also significantly higher than that of its mutant (pFTLTR1E) in which 2 of the 3 enhancers were deleted in FT-1 and Jurkat cells. Both of these differences were not observed in a feline fibroblastic cell line (CrFK). Moreover, mutations affecting the consensus motifs for LVb, SV40, NF 1, GRE and FLV-1 resulted in decreased basal activity of the FeLV LTR (pFTLTR1E) in FT-1, Jurkat and CRFK cells. The decrease of the promoter activity was especially remarkable in FT-1 cells. The present study revealed the strong promoter activity of the FeLV LTR with 3 enhancer repeats and its modular enhancer elements positively regulating the transcription in a relatively tissue specific manner. PMID- 9209340 TI - To be or not to be a T-lymphomas, that is determined by a dominant gene Tlsm-1 in mouse models. AB - A single dominant gene Tlsm-1 was found to determine the type of spontaneous lymphomas to be T in the cross between AKR/Ms and SL/Kh. Microsatellite analysis mapped Tlsm-1 at the position 61 cM from centromere of Chr. 7. Study of this segment of T-lymphoma prone AKXD Rl strains also showed association of Tlsm-1 with T-lymphomas. On the other hand, lymphoma latency was significantly shorten by a recessive gene lla of SL/Kh. By a quantitative trait analysis, lla was located in class II gene in MHC. PMID- 9209342 TI - Apoptosis in peripheral CD4+T cells and thymocytes by Marek's disease virus infection. AB - Histological study revealed that Marek's disease virus (MDV) can cause apoptosis in peripheral blood lymphocytes (PBL) in latently infected chickens. Analysis of DNA fragmentation indicated that CD4+T cells but not CD8+T cells underwent apoptosis. These apoptotic changes were also observed in the thymus during the acute phase of the infection. Flow cytometry analysis showed the drastic decrease of CD4+CD8+ thymocytes, indicating that MDV can induce apoptosis in CD4+CD8+ immature thymocytes in acutely infected chickens. These changes might be involved in the immuno-suppression induced by MDV. PMID- 9209341 TI - Immunosuppression by metastatic lymphoma derived altered retroviral gp70 molecule. AB - In this report we describe the characteristics of an immunosuppressive molecule from an Abelson Leukemia Virus transformed highly malignant and metastatic RAW117 H10 murine large cell lymphoma cells. Our studies have shown that the mitomycin-c treated or irradiated RAW117-H10 cells very significantly (p < 0.001) inhibited the nitrogen induced proliferation of normal Balb/c splenocytes. The cell surface extracts from the immunosuppressive RAW117-H10 lymphoma cells significantly inhibited the in vitro T cell or NK/LAK cell functions. Our in vivo studies demonstrated that there was a significant suppression of immune response in the Balb/c mice bearing RAW117-H10 cells when compared with mice bearing low metastatic parental RAW117-P cells or control mice. Subsequently we isolated and purified the main molecule responsible for this immunosuppression and found that the molecule is a glycoprotein with a molecular weight of 70 kD with an isoelectric point of 4.3, which cross reacted with antibodies to murine leukemia virus envelope gp70 molecules. Further analysis using immunoelectrophoresis, molecular probing techniques, and mannose specific lectin binding assay revealed that the immunosuppressive 70 kD molecule, is different from the wild type MLV envelope gp70 molecule and thus appears to be an altered gp70 molecule. These studies demonstrate that the metastatic lymphoma associated immunosuppression may facilitate the growth and metastasis of tumor cells. Our results also elucidate the possible mechanism(s) of retrovirus induced immunosuppression and the molecular basis of this retroviral envelope-mediated process in viral pathogenesis and tumor progression. PMID- 9209343 TI - The role of E-selectin for neutrophil activation and tumor metastasis in vivo. AB - The selectin class of adhesion molecules plays a critical role in facilitating leukocyte adhesion to and subsequent transmigration of endothelium. On this basis, selectins have been suggested to promote metastasis of certain types of tumors, although direct evidence is lacking. To explore this hypothesis two sets of transgenic mice were developed, one TgNES, which constitutively expresses cell surface E-selectin in all tissues, the other TgNEsol, which expresses truncated, soluble E-selectin in the liver. Both transgenic mice showed apparently normal phenotype. However in TgNES mice, but not in TgNEsol mice, the number of neutrophil decreased to 50% compared with that in their negative littermate. And also these neutrophils were markedly activated. On the other hand, B16 F10 melanoma cells were stably transfected with alpha 1-3/4 fucosyltransferase specific cDNA (B16F10 ft), allowing them to express E-selectin ligands. Normal mice injected with B16F10 ft developed lung tumors exclusively. In contrast, TgNES mice developed massive, infiltrating liver tumors. TgNEsol mice also developed liver tumors that grow more slowly. These observations suggest the important role of E-selectin for neutrophil activation and tumor metastasis in vivo. PMID- 9209344 TI - Augmentation with serum thymic factor of suppressive effects on retroviral tumor development in hosts immune to v-onc product. AB - Inbred adult female rats were immunized against syngeneic ST-FeSV induced sarcoma cells. ST-FeSV was injected subcutaneously into 57 neonates (vaccinated) born from these immunized females and into 60 non-vaccinated syngeneic neonates. Serum thymic factor (FTS) was injected subcutaneously into 10 of vaccinated and 30 of non-vaccinated rats. Sarcomas developed in 40.4% (19/47) of vaccinated (A), 20.0% (2/10) of vaccinated FTS injected (B), 63.3% (19/30) of non-vaccinated FTS injected (C), and 76.7% (23/30) of non-treated (D) rats. By AB immunostaining using antibody to v-fes product (P85), sarcomas developed in 10 of 13 rats of group C tested, and 3 of 6 rats of group D tested were positive, but those in 7 rats of group A and 2 rats of group B tested were all negative. Lung metastasis was observed in rats of all groups except those of B group. All sera of animals that developed sarcomas were positive to P85 in Western blot analysis. These results showed that FTS augmented suppressive effects on sarcoma development in hosts immune to the viral oncogene product. PMID- 9209345 TI - The role of tumor-associated antigen in bovine leukemia virus-induced lymphosarcoma. AB - Bovine leukemia virus (BLV) is associated with enzootic bovine leukosis (EBL), which is the most common neoplastic disease of cattle. To clarify the way in which BLV-infected cattle progress from the asymptomatic stage to the lymphoma stage, we produced a monoclonal antibody (MAb) c143 which recognized a tumor associated antigen (TAA) that is phosphorylated in the transformed state of BLV infected B-lymphoid cells. Since the nature of c143 TAA was likely to be that of the major histocompatibility complex (MHC) class II antigens, we isolated cDNAs for bovine MHC (BoLA) class II a-chains and b-chains, produced transfectants that expressed a single type of BoLA class II molecules and analyzed them by flow cytometry with c143 MAb. The c143 MAb recognized the transfectant expressing BoLA DR but not BoLA-DQ. However, the treatment of lymphocytes with c143 or anti-BoLA DR MAb induced different effects. Although mixed lymphocyte reaction (MLR) was inhibited by the addition of anti-BoLA-DR MAb, the c143 MAb did not inhibit a proliferative response of T cells in MLR. Increased spontaneous proliferation of lymphocytes in healthy donors was obtained in the presence of c143 MAb but not anti-BoLA-DR MAb, and was much in lymphocytes from the carrier. Moreover, the patterns of immunohistological staining for c143 MAb in BLV-infected sheep showed distinguishing differences from those of anti-BoLA-DR MAbs. PMID- 9209346 TI - Levels and role of cytokines in bovine leukemia virus (BLV) infection. AB - Bovine leukemia virus belongs to a small subfamily of exogenous retroviruses that includes the human retroviruses HTLV-1, HTLV-II and the simian virus, STLV-1. Like other retroviruses, infection with BLV results in deregulation of the host immune system at both humoral and cellular levels. An approach which might help in the elucidation of some immune impairment phenomena is the investigation of the role that cytokines play in the pathogenesis and immune response of BLV infected animals. Here we describe our findings on IL-6 and TNF. We have found that the levels of IL-6 in the sera of BLV infected cows which show persistent lymphocytosis (BLV+ PL+) were significantly higher than those of BLV infected with no lymphocytosis (BLV+ PL-) or BLV negative cows (BLV-). The same results were obtained by measuring the spontaneous production of IL-6 in peripheral blood mononuclear cells (PBMC). Furthermore, PBMC derived from BLV+PL+ cows secrete higher levels of IL-6 and TNF alpha than those derived from BLV+PL- and BLV- ones following in vitro exposure to the BLV gp51 antigen, bacterial endotoxins (LPS) and ConA. Similar results were obtained when supernatants from stimulated adherent (monocytes, macrophages) and non-adherent cells (B and T lymphocytes) were tested. When exogenous IL-6 and TNF alpha were added to BLV infected cells in vitro, the expression of viral antigens was strongly suppressed. Thus, the possibility exists that the elevated production of IL-6 and even more than that of TNF alpha play a role as contributing factors to the latency of the clinical expression in BLV infection. PMID- 9209347 TI - Tumor-associated antigen in lymph nodes during the progression of enzootic bovine leukosis. AB - A monoclonal antibody, (MAb) c143, that recognizes a tumor-associated antigen (TAA) that is upregulated on neoplastic B cells in cattle with enzootic bovine leukosis (EBL), was used as a marker to study disease progression. Immunohistochemical examination of neoplastic tissue and lymph nodes from animals with EBL, revealed three morphologically definable stages of change in the architecture of lymph nodes associated with infiltration and proliferation of neoplastic cells: 1) presence of c143 positive cells at the marginal sinus with no apparent changes in lymph node architecture at the earliest stage of neoplastic cell accumulation, 2) presence of positive cells extending into and distorting the architecture of the lymph node with clear evidence of proliferation, and 3) presence of positive cells throughout the lymph node with total disruption of lymph node architecture. The results indicated that, at the earliest stages of EBL, neoplastic cells in peripheral blood may accumulate at the marginal sinus area and subsequently proliferate and infiltrate pressing follicles leading to the development of clinical signs of lymphosarcoma. PMID- 9209348 TI - Peptide-based bovine leukemia virus (BLV) vaccine that induces BLV-Env specific Th-1 type immunity. AB - In controlling retrovirus infection and replication, cell-mediated immunity (CMI) is considered to be effective. To develop a synthetic peptide vaccine capable of inducing CMI, mannan-coated liposome encapsulating 20-mer synthetic peptide, spanning the 98-117 amino acids of bovine leukemia virus (BLV) envelope glycoprotein (Env) gp51 was constructed and inoculated to BALB/c mice. The liposome induced specific delayed-type hypersensitivity, lymphocyte proliferative responses, and a weak cytotoxic lymphocyte response. The spleen cells from the immunized mice produced a large amount of IFN-gamma and IL-2, whereas they released neither IL-4 or IL-10. Mannan-coated liposome containing two kinds of peptides (the 121-140 and 142-161 regions of BLV Env gp51) also induced peptide specific lymphocyte proliferative response and IFN-g production in C57BL/6 mice. Thus, the synthetic T-cell epitope peptide-liposome system augmented a strong Th 1 type immunity in mice. PMID- 9209349 TI - Restriction of Friend virus-induced erythroid cell proliferation by CD4+ T lymphocytes that recognize a single gp70 epitope. AB - Friend murine retrovirus complex induces acute and fatal erythroleukemia when inoculated into immunocompetent adult mice. The development of leukemia after inoculation of Friend virus complex is controlled by several host genes. Some of the host genes influence immune responses against the viral antigens. Both CD4 positive T helper cells and CD8-positive cytotoxic T-lymphocytes specific for Friend viral antigens are required for spontaneous resistance against the virally induced leukemia. We have identified two separate T helper cell epitopes in the gp70 envelope glycoprotein encoded by the helper component of Friend virus complex. Immunization of mice with a synthetic peptide that represented one of the two T helper cell epitopes by a single injection with an adjuvant induced potent protective immunity against Friend virus-induced leukemia, even in the absence of CD8-positive T lymphocytes. In the immunized mice, virus-infected erythroid progenitor cells were rapidly eliminated from the spleen within two weeks after inoculation of the Friend virus. These data indicate unexpected importance and efficacy of CD4-positive T helper cells in immunity against retrovirus infections. PMID- 9209350 TI - Cell-free transmission of Fv-4 resistance gene product controlling Friend leukemia virus-induced leukemogenesis in mice. AB - Fv-4 is a mouse gene that dominantly confers resistance to infection by ecotropic murine leukemia virus (MuLV). The Fv-4' env antigen that binds to the cell surface of Fv-4'-bearing C3H cells was found in sera from normal Fv-4'-bearing C4W mice. The serum Fv-4' env antigen binds to ecotropic MuLV receptors, shown by specific binding to transfectant cells expressing ecotropic MuLV receptors but not to parental mink cells. To determine whether the binding of Fv-4' env antigen to the putative MuLV receptors would block FLV infection, C3H thymocytes or spleen cells that had been preincubated with C4W serum were mixed with FLV and the subsequent production of MuLV specific antigens was examined. C3H thymocytes or spleen cells treated with C4W serum became refractory to binding by FLV. These results provide evidence that the Fv-4' env antigen is released from C4W-derived cells in vivo and binds to cells expressing surface receptors for ecotropic MuLV, thereby protecting them from infection with FLV. The implication of these findings for gene therapy of retrovirus-induced disease such as acquired immune deficiency syndrome (AIDS) is discussed. PMID- 9209351 TI - Molecular mechanism for retroviral neuropathogenesis: possible involvement of capillary endothelial cells. AB - A neuropathogenic variant of Friend MuLV, PVC-211, causes rapidly progressive spongiform neurodegeneration in susceptible rats and mice. Major targets of PVC 211 MuLV infection are brain capillary endothelial cells (BCEC), suggesting that virus-infected BCEC may play crucial roles in neurological disease induction. Consistent with this possibility, studies using chimeric viruses constructed between PVC-211 MuLV and non-neuropathogenic Friend MuLV have revealed that the BCEC tropism of the virus correlates with its neuropathogenicity. Possible involvement of cytokine expression by PVC-211 MuLV-infected BCEC in the induction of neuropathological changes will be discussed. PMID- 9209352 TI - Factors affecting induction of neurological disorder in mice by PVC viruses and the sequence of env-LTR region of PVC441. AB - PVC111, PVC211, PVC321 and PVC441 cause neurological disorders associated with tremor and paralysis in rats. We tested the pathogenicity of these viruses in mice. Although histopathological studies revealed spongiform degeneration in the spinal cords of NFS mice infected with each PVC virus, only PVC441 caused a high incidence of tremor and paralysis. Further studies with PVC441 revealed dose and age dependence for tremor induction. In contrast to NFS mice, BALB/c, DBA/2 and C57BL/6 mice infected with PVC441 virus showed no neurological symptoms, although the virus replicated in each strain to titers within 5-fold of the titer in NFS mice. Despite absence of neurological symptoms, high degree of neuronal degeneration in the lumbar spinal cord was found in PVC441-infected BALB/c mice. Low degree of neuronal degeneration was found in PVC441-infected DBA/2 or C57BL/6 mice. Genetic crosses of these resistant mice with susceptible NFS mice indicated that resistance to PVC441-induced tremor induction was dominant in all strains and suggested that various host genes may control the susceptibility of mice to tremor induction by PVC441 virus. Sequencing of env-LTR region of PVC441 revealed an intermediate character between PVC211 and F-MuLV. PMID- 9209353 TI - Cooperating events in lymphomagenesis mediated by feline leukemia virus. AB - Feline leukemia virus (FeLV)-mediated lymphomagenesis in the domestic cat has been examined as a model of lymphoid malignancy in a naturally outbreeding population. The pathogenesis of two distinct, naturally occurring types of FeLV induced tumors has been investigated: (1) a thymic lymphoma of T-cell origin, typical of FeLV-induced lymphoma, and (2) an extrathymic, extranodal lymphoma of non-B non-T-cell origin. The genetic features of these tumors are clearly distinguishable, and include determinants encoded both by the virus and the host. Virally encoded determinants of pathogenesis include the long terminal repeat (LTR) and the envelope SU protein. Cellular determinants include the involvement of a set of proto-oncogenes, and other factors characteristic of the specific cell type of origin of the tumor. Functional studies are aimed at evaluating the action and interaction of these genetic determinants in the pathogenesis of lymphoma in an animal model system. PMID- 9209354 TI - Models of HTLV-I-induced diseases. Infectious transmission of HTLV-I in inbred rats and HTVL-I env-pX transgenic rats. AB - To examine the pathogenic roles of HTLV-I in HTLV-I-induced diseases, we developed two models; namely HTLV-I carrier rats and HTLV-I env-pX transgenic rats. Among life long HTLV-I carriers in seven rat strains, only WKAH rats with the RT1k haplotype developed chronic progressive myeloneuropathy, resembling HAM/TSP clinically and histologically in humans, designated as HAM rat disease and after long incubation periods. Apoptosis of myelin forming cells, oligodendrocytes and Schwann cells associated with HTLV-I infection appears to be the primary cause of HAM rat disease. Local activation of the pX gene and TNF alpha gene was evident in these rats. WKAH rats transgenic for HTLV-I env-pX gene were established and at age 5 weeks, swelling of the bilateral ankle joints began to develop and histological features of the affected joints resembled findings in cases of rheumatoid arthritis (RA): high-titers of rheumatoid factors were present in these rats. A series of vascular collagen diseases such as polyarteritis nodosa-like angiitis, polymyositis, myocarditis, and Sjogren's syndrome-like sialodenitis together with RA were present, even in one individual animal. These transgenic rats as well as HAM rats appear to be suitable animal models for elucidating pathogenic mechanisms implicated in HTLV-I-induced diseases and also various demyelinating vascular collagen diseases of unknown etiology. PMID- 9209356 TI - Constitutive activation of Stat-related DNA-binding proteins in erythroid cells by the Friend spleen focus-forming virus. AB - The erythroleukemia-inducing Friend spleen focus-forming virus (SFFV) encodes a unique envelope glycoprotein which allows erythroid cells to proliferate and differentiate in the absence of the erythroid hormone erythropoietin (Epo). In an attempt to understand how the virus alters the growth of erythroid cells, studies were carried out to determine if virus infection leads to the constitutive activation of the Jak-Stat pathway, one of the signal transduction pathways activated by Epo. Our data indicates that expression of SFFV in erythroid cells leads to the constitutive activation of the same Stat proteins that are transiently activated by Epo. While constitutive activation of Stat proteins by SFFV is associated with Epo-independent proliferation of splenic erythroid progenitor cells from Fv-2-sensitive mice and Epo-dependent HCD-57 cells, it is not sufficient to induce their differentiation. Although constitutive activation of the same Stat proteins is detected in erythroid cells from SFFV-infected Fv-2 resistant mice, it does not lead to their Epo-independent growth. It is also not required for transformation of erythroid cells by SFFV. Studies are in progress to identify the mechanism by which Stat proteins are phosphorylated in SFFV infected cells in the absence of Epo. Although it has been shown that Epo activates Stat proteins through Jak2 kinase, our results suggest that the SFFV induced Stat protein activation is Jak2-independent. PMID- 9209355 TI - MuLV-insertional mutagenesis of c-myb and Mml1 in a murine model for promonocytic leukemia. AB - Analysis of retroviral integration sites in MuLV-induced promonocytic leukemias has determined that two genetic loci, c-myb and Mml1, can contribute to disease development but not in the same leukemia. Recent studies aimed at understanding the function of Myb in leukemia development have focused on the consequences of ectopic Myb expression on monocytic and granulocytic differentiation in vitro. In all instances Myb was shown to block growth arrest but not commitment to differentiation, a result which is consistent with observed effects of Myb in leukemia development. No effect of Myb protein truncation was observed in these studies although similar truncations are produced as a result of insertional mutagenesis. Common integration site, Mml1, was recently identified and mapped to mouse chromosome 10 within 1cM of c-myb. Despite its linkage to c-myb, Myb mRNA and protein expression appear to be unaffected in leukemias with Mml1 integrations. PMID- 9209358 TI - HTLV-I env-pX transgenic rats: prototype animal model for collagen vascular diseases. AB - To evaluate the function of HTLV-I env-pX gene in vivo, we developed two lines of transgenic rats (env-pX rats) that expressed env-pX gene products, under control of own LTR promotor. In various tissues of the rats, env and pX mRNAs were constitutively expressed, irrespective of age. At age 5 weeks, swelling of the bilateral ankle joints histologically showing synovial lining hyperplasia, severe chronic inflammation, erosion of the joint cartilage, and bone destruction with pannus formation began to develop in these env-pX rats. These histologic features resemble those of rheumatoid arthritis (RA) in man. High titered rheumatoid factors and low anti-dsDNA antibodies and hyper-gamma globulinemia were detected. Necrotizing arteritis resembling polyarteritis nodosa, polymyositis, myocarditis and Sjogren syndrome-like sialoadenitis developed, together with RA-like arthritis even in one individual animal. Thymic atrophy with low body weight was also observed. The evidence indicates that env-pX rats appear to be suitable animal models for elucidating pathogenetic mechanisms involved in not only HTLV-I related diseases but also various collegen vascular and autoimmune diseases of unknown etiology in man. PMID- 9209357 TI - HTLV-I induced myeloneuropathy in WKAH rats: apoptosis and local activation of the HTLV-I pX and TNF-alpha genes implicated in the pathogenesis. AB - To investigate the pathogenesis of HTLV-I associated diseases, we established a rat model for HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP) in WKAH rats. In the spinal cords of WKAH rats carrying HTLV-I, chronological histopathology revealed the occurrence of apoptotic cell death starting at 9 months after the infection, followed by demyelination, macrophage infiltration, and the activation of astrocytes starting at 12, 15 and 20 months, respectively. Apoptosis of the Schwann cells was also observed in the peripheral nerves of these rats. By RT-PCR, pX mRNA of HTLV-I was selectively expressed in the diseased spinal cords and peripheral nerves, but not in the unaffected cerebra, cerebella, even though provirus DNAs were consistently identified in these tissues. Among several cytokines examined, mRNA expression and production of TNF alpha were frequently detected in the spinal cord and the cerebrospinal fluid. The collective evidence suggests that the selective activation of HTLV-I, in particular Tax expression, and/or the production of TNF-alpha in target spinal cord and peripheral nerves are causally related to apoptotic death of the oligodendrocytes and Schwann cells, a major pathogenetic pathway of HTLV-I induced myeloneuropathy in the WKAH rat. PMID- 9209359 TI - Establishment of HTLV-1 carrier mice by injection with HTLV-1-producing T cells. AB - We injected with HTLV-1-producing T-cells, MT-2, into C3H/HeJ and Balb/c mice intraperitoneally within 24 hours after birth and at one week old. At 3 months old, HTLV-1 provirus was detected in peripheral blood mononuclear cells in both mice. It was also detected in lymph nodes, thymus, spleen and liver of those mice. The antibody response against HTLV-1 Gag antigen was detected in some of Balb/c mice. These findings suggest that the C3H/HeJ and Balb/c mice were infected with HTLV-1 persistently. The HTLV-1-infected mice should be helpful for studying the pathological state of HTLV-1 carriers and for an establishment of a small animal model of HTLV-1 related diseases including ATL. PMID- 9209360 TI - A new retroviral AIDS-like disease and/or acute leukemia induced in mice by alloimmune stimuli. AB - We have shown a new phenomenon demonstrating that BALB/c female mice mated to C57BL/6 males during a year (7-10 pregnancies) develop AIDS-like disease or acute leukemia after an additional immunization with fixed ConA activated paternal (C57BL/6) lymphocytes. The AIDS-like disease is sexually and vertically transmissible and easily transferable to intact BALB/c and C57BL/6 mice by filtered plasma of affected animals. PMID- 9209361 TI - Genetic factors predisposing to B-CLL and to autoimmune disease in spontaneous murine model. AB - The frequent occurrence of autoimmune diseases in patients with B cell chronic lymphocytic leukemia (B-CLL) or in their family members suggests the involvement of related predisposing genetic factors in the two distinct disorders. Because the majority of B-CLL is of CD5 B cell type, and because the majority of CD5 B cells produces polyreactive autoantibodies, certain regulatory abnormalities in the proliferation and differentiation of CD5 B cells appear to be involved in B CLL and autoimmune disease, respectively. In studies using MHC(H-2)-congenic New Zealand mouse strains, NZB(H-2d), NZW(H-2z) and NZB x NZW F1(H-2d/z), we found that while H-2d/z heterozygosity acts as one genetic predisposing element for autoimmune disease, by providing the element for abnormal differentiation of CD5 B cells, the H-2z/z homozygosity serves as one predisposition for B-CLL, by promoting the abnormal proliferation of CD5 B cells. Another non-MHC-linked genes were also involved in the aberrant proliferation of CD5 B cells, as determined by microsatellite DNA analysis. Among these, there was a single dominant NZW locus located on chromosome 6, closely linked to the locus for TNF receptor p55. Hence our mouse model provides an appropriate tool for studying the relationship between genetic factors predisposing to B-CLL and autoimmune diseases. PMID- 9209362 TI - Chromosomal translocations and leukaemia: a role for LMO2 in T cell acute leukaemia, in transcription and in erythropoiesis. AB - The LMO2 gene associated with T cell acute leukaemia has been used as an example of a gene activated by association with the T cell receptor genes after chromosomal translocations. The gene is shown to encode a LIM protein which is involved in protein interactions and during normal haematopoiesis is necessary for erythroid development. LMO2 has been shown to cause tumours when aberrantly expressed and to be able to heterodimerise with TAL1 to facilitate tumour development. PMID- 9209364 TI - The runt protein and its companion PEBP2: a close link between this transcription factor and AML. PMID- 9209363 TI - Rearrangements of the AML1/CBFA2 gene in myeloid leukemia with the 3;21 translocation: in vitro and in vivo studies. AB - AML1 is involved at the breakpoint of chromosome 21 band q22 in several recurring chromosomal translocations associated with myeloid and lymphoid leukemias. AML1 corresponds to CBFA2, and encodes one of the DNA-binding subunits of the enhancer core binding factor CBF. Other members of this family of DNA-binding proteins are CBFA1 and CBFA3, also known as AML3 and AML2. The three proteins are characterized by a highly conserved domain (runt domain, > 90% homology) at the amino end that is necessary for DNA-binding and protein dimerization, and by a unique domain at the carboxyl end that is necessary for transactivation. Two recurring chromosomal translocations involving AML1 associated with myeloid leukemias are the t(8;21)(q22;q22), seen in 20% of patients with acute myeloid leukemia (AML) M2, and the t(3;21)(q26;q22), that occurs in myeloid leukemias primarily following treatment with topoisomerase II inhibitors. In five patients with a t(3;21) whom we studied, AML1 is interrupted by the translocation breakpoint between the runt domain and the transactivation domain, and is fused to two genes on chromosome band 3q26: EAP, which encodes the ribosomal protein L22, and MDS1, which encodes a small polypeptide of unknown function. In one of the five patients we studied, a fusion with a third gene EVI1 also occurs. The fusion of EAP to AML1 is not in frame, and leads to a protein that is terminated shortly after the fusion junction by introduction of a stop codon. The fusion of AML1 to MDS1 is in frame, and adds 127 codons to the interrupted AML1. Thus, in the five cases that we studied, the 3;21 translocation results in expression of two coexisting chimeric mRNAs which contain the identical runt domain at the 5' region, but differ in the 3' region. In addition, the chimeric junction AML1/MDS1/EVII has been detected in cells from one of our patients with the 3;21 translocation. Several genes necessary for myeloid lineage differentiation contain the target sequence for AML1 in their regulatory regions. We have compared the normal AML1 to AML1/MDS1 and AML1/EAP as transcriptional regulators of the CSF1R promoter which contains the CBF target sequence. Our results indicate that whereas the normal AML1 can activate the promoter, the chimeric proteins compete with the normal AML1 and repress expression from the CSF1R promoter. To determine the role of the chimeric proteins in cell growth, we expressed their cDNA in rat fibroblasts. When either fusion gene is expressed, the cells lose contact inhibition and form foci over the monolayer. However, only cells expressing AML1/MDS1 grow as large tumors in nude mice. Thus, although both chimeric genes have similar effects in transactivation of the CSF1R promoter, they affect cell growth as tumor promoters differently in vivo. PMID- 9209365 TI - Detection of T-cell receptor delta gene rearrangement by clonal specific polymerase chain reaction. AB - A sensitive and specific technique to detect minimal residual disease for T-cell malignancies was explored. Southern analysis and polymerase chain reaction (PCR) were used to detect the rearranged V-D-J segment of T-cell receptor delta (TCR delta) gene from malignant cell specimens of patients with leukemia and lymphoma of T-cell lineage. The PCR product was sequenced and from the DNA sequences of the V-D-J region, a 3' antisense primer was designed and synthesized for clonal specific PCR (CS-PCR). Seven of the 22 T-ALL (32%) and 5 of 18 (28%) T-cell lymphoma showed clonal rearrangement by Southern analysis. Six of the 7 (86%) T ALL and 4 of the 5 (80%) T-cell lymphoma which were Southern positive were also positive by TCR delta PCR. The PCR products of four cases of T-ALL showing clonal pattern by TCR delta PCR amplification were successfully sequenced and CS-PCR amplification performed. CS-PCR detected with confidence specific clonal rearrangement in a mixture containing 0.003% of malignant cells. Marrow specimens obtained at diagnosis and subsequent follow-ups from the 4 T-ALL patients were studied by Southern analysis, TCR delta PCR and CS-PCR. The first patient was in continuous morphological complete remission for more than 3 years and had persistently negative Southern, TCR delta PCR and CS-PCR results on follow-up. Initial follow-up marrow samples from the second patient had persistently positive CS-PCR results while they were still morphologically and TCR delta PCR negative and the patient had a frank leukemic relapse at 18 months. The other two patients had persistent disease by conventional morphological examination, Southern analysis, TCR delta PCR and CS-PCR studies were all positive as expected. CS-PCR is a highly specific and sensitive technique in detecting minimal residual disease for T-cell malignancies. Its potential applications warrant further clinical evaluation and correlation. PMID- 9209366 TI - Nuclear proteins binding to the recombination hotspot region of the retinoic acid receptor alpha gene. AB - Acute promyelocytic leukemia (APL) has been characterized by 15;17 chromosomal translocation, which involves the retinoic acid receptor alpha (RARA) gene on chromosome 17 and the PML gene on chromosome 15. An extremely restricted region (ERR) of 50 bps within the second intron of the RARA gene was identified as the cluster region of breakpoints by sequencing analyses. ERR was tested by in vitro transfection-recombination assay, and was shown to be the recombination hot spot. In this study, presence of DNA binding proteins to the 148 bps DNA fragment which contains ERR was confirmed by gel-mobility shift analysis in the nuclear extract of NIH3T3 cells and human leukemia cell lines. Furthermore, in vitro study with the mouse sarcoma cell lines using the recombination reporter plasmid containing ERR showed that ERR might be involved in the homologous recombination in addition to the illegitimate recombination. The DNA binding proteins specific to ERR might play an important role in chromosome translocation. PMID- 9209367 TI - Molecular analysis of the t(15;17) translocation in de novo and secondary acute promyelocytic leukemia. AB - To study mechanism of chromosomal translocation, we analyzed the breakpoints (b/p) of the PML and RARA genes in 120 and 5 patients with de novo and secondary (therapy-related) acute promyelocytic leukemia (APL), respectively. In de novo APL, the b/p in the PML gene were clustered in introns 3 (bcr 3; 30%) and around intron 6 (bcr 1 and 2: 70%). The b/p of the RARA gene were widely distributed in intron 2. In studied 8 de novo APL patients, no consensus sequence-motif was found around the b/p, but there were identical stretches of one to seven nucleotides between the PML and RARA genes in the joining regions, suggesting non selective DNA double strand cleavage followed by single strand base-pairing within identical short stretches as a molecular mechanism of the translocation. In 4 secondary APL patients after chemotherapy including etoposide against Langerhans cell histiocytosis, the b/p of the PML gene were located in intron 6, and those of the RARA gene were in a restricted region within intron 2, 1 kb EcoRI-BamHI fragment, while in an APL patient after chemotherapy without etoposide against breast cancer, the b/p of the PML and RARA genes were located in intron 6 and another region within intron 2, respectively. These data suggest that a different mechanism was associated with the t(15;17) translocation in etoposide-related APL. PMID- 9209368 TI - Disruption of T-cell differentiation precedes T-cell tumor formation in LMO-2 (rhombotin-2) transgenic mice. AB - Transgenic mice expressing LMO-2 (rhombotin-2) were constructed by placing the LMO-2 gene under control of the metallothionein promoter. Thymic tumors developed in approximately 15% of the transgenic mice between 37 and 71 weeks. Only T-cell tumors were found in the transgenic mice despite high expression of LMO-2 in all tissues. The thymic tumors were aggressive and were invariably associated with metastasis to non-lymphoid organs. In approximately 50% of apparently healthy transgenic mice there was up to a 10-fold expansion of CD4-CD8- double negative (DN) cells. Expansion of the DN cells was accompanied by the compensatory decrease in CD4+CD8+ double positive (DP) cells, indicating that breach of homeostasis within the thymus had not occurred in these animals. The increase in DN cells was associated with a clonal expansion of thymocytes, and increased proliferation within the thymus. Our data indicate that the ectopic expression of LMO-2 in T-cells disrupts normal T-cell differentiation by selectively expanding the DN thymocyte population prior to breach of homeostasis and overt leukemia/lymphoma. PMID- 9209369 TI - Rapid detection of lymphoma-specific translocations in interphase nuclei of non Hodgkin's lymphoma by fluorescence in situ hybridization. AB - We have recently developed a method to detect tumor-specific rearrangement of the IgH gene in interphase nuclei by fluorescence in situ hybridization. Tumor specific IgH gene rearrangement is equivalent to 14q32.33 translocation. Using this approach, we detected 14q32.33 translocation in 29 of 70 patients with B cell non-Hodgkin's lymphoma (NHL). Chromosome t(3;14) was found in 10 of these 29 patients, and were demonstrated as a fusion signal of BCL6 and VH gene probes in interphase nuclei. Furthermore, in another series of 11 patients and a NHL cell line, we demonstrated t(14;18) and t(11;14) in interphase and metaphase cells with a combination of BCL2 (or PRAD1) with IgH gene probes. Interphase FISH with lymphoma-associated gene probes is a rapid procedure for cytogenetic diagnosis of B-cell NHL. PMID- 9209370 TI - Leukemogenesis by the chromosomal translocations. AB - The AML1 is the most commonly involved transcription factor gene in human leukemias and forms chimeric transcription factor genes, namely, AML1/MTG8 by the t(8;21), AML1/EVI-1 by the t(3;21), and TEL/AML1 by the t(12;21). The AML1a and AML1b, two isoforms of the AML1 protein, are translated from the AML1 gene by the alternative splicing. The shorter form and longer form are named as AML1a and AML1b, respectively. The AML1b contains the runt homology domain as a DNA-binding domain and the serine/proline/threonine-rich domain (PST domain) as a putative transactivation domain. The AML1a contains only the runt homology domain, but not the PST domain. The AML1b has a transactivation ability through the PEBP2 site and stimulates differentiation of myeloid cells. On the other hand, AML1a cna bind the PEBP2 site, but does not show a transactivation ability through the PEBP2 site. The AML1a dominantly suppresses transactivation induced by the AML1b and has the ability to block differentiation of myeloid cells. It is a reasonable hypothesis that the molecular ratio of AML1a to AML1b in myeloid cells could determine whether they proliferate or undergo a terminal differentiation. PMID- 9209372 TI - An acute myeloid leukemia gene, AML1, regulates transcriptional activation and hemopoietic myeloid cell differentiation antagonistically by two alternative spliced forms. AB - The AML1 gene on chromosome 21 is disrupted in the (8;21)(q22;q22) and (3;21)(q26;q22) translocations associated with myelogenous leukemias and encodes a DNA-binding protein. From AML1 gene, two representative forms of proteins, AML1a and AML1b, are produced by an alternative splicing. Both forms have DNA binding domain, but AML1a lacks a putative transcriptional activation domain which AML1b has. Here we demonstrate that AML1a, which solely has no effects as a transcriptional regulator, dominantly suppresses transcriptional activation by AML1b, and that AML1a exhibits the higher affinity for DNA-binding than AML1b. Furthermore a dominant negative form of AML1, AML1a, totally suppressed granulocytic differentiation otherwise induced by granulocyte colony-stimulating factor when AML1a was overexpressed in 32Dc13 murine myeloid cells. Such differentiation block by AML1a was canceled by the concomitant overexpression of AML1b. These data strongly suggest that a transcriptionally active form of AML1 is essential for the myeloid cell differentiation. In addition, we observed an altered expression level of AML1 along with the myeloid differentiation in several hemopoietic cell lines. In these cases, at least, the AML1 expression level is a potential regulator for myeloid cell differentiation. PMID- 9209373 TI - A 720-kb yeast artificial chromosome contig spanning human chromosome 5q31.3-32. PMID- 9209371 TI - Significance of MTG8 in leukemogenesis. AB - MTG8 is a counterpart gene of AML1 in acute myeloid leukemia with t(8:21) translocation. Most of the coding region of the MTG8 is fused with AML1 runt domain. In normal tissues, the MTG8 is highly expressed in brain, but not in hematopoietic tissues. MTG8 may be important in leukemogenesis as well as in AML1 truncation. The function of MTG8 is assumed to be as a transcription factor, because it possesses several features common to transcription factors; putative zinc finger motifs, serine/threonine/proline-rich sequences and a region similar to TAF110. In this paper, we report on the protein properties of the MTG8. PMID- 9209375 TI - A functional screen for regulators of the c-Abl protein tyrosine kinase. AB - c-Abl protein tyrosine kinase activity is tightly regulated in vertebrate cells. Several mutations can activate Abl and convert it into an oncogene. In man, chromosomal translocations result in fusion proteins associated with chronic myelogenous leukemias and some acute lymphocytic leukemias. In viral forms of abl, gag sequences are fused to Abl portions resulting in a deletion of N terminal sequences. To study c-Abl activity in a cellular environment likely to lack specific regulators, we have expressed human c-Abl in Schizosaccharomyces pombe in an inducible fashion. c-Abl causes growth arrest followed by death of the cells. Mutations in the SH2 domain or in the autophosphorylation site dramatically reduce the ability of Abl to confer the growth arrest phenotype and to phosphorylate endogenous proteins, suggesting a fundamental role of these structures in the activity of the enzyme. An SH3 domain deletion mutant of Abl is as active as c-Abl in yeast indicating that there is no intrinsic regulation of c Abl occurring via the SH3 domain and suggesting that the inhibitory effect of the SH3 domain observed in cells of vertebrate origin is mediated by a factor that is absent in fission yeast. We have used this assay to functionally screen a human cDNA library for molecules able to counteract the lethal effect of c-Abl expression. We are currently in the process of characterising the isolated clones. We hope to identify among them the molecule(s) responsible for regulating c-Abl activity in human cells. PMID- 9209376 TI - Long distance polymerase chain reaction for detection of chromosome translocations in B-cell lymphoma/leukemia. AB - To establish a rapid and sensitive method to detect neoplastic cells carrying a specific chromosomal translocation in B-cell lymphoma/leukemia, we have developed a novel strategy based on long distance polymerase chain reaction (LD-PCR) amplification. Genomic DNA were extracted from tumor cells carrying a t(14;19)(q32;q13), a t(8;14)(q24;q32), a t(3;22)(q27;q11), a t(2;3)(p12;q27), and a t(3;14)(q27;q32). Oligonucleotide primer pairs were designed to be complementary to exons or flanking sequences of the BCL3, c-MYC and BCL6 oncogenes, and to constant region genes of the IG genes. LD-PCR with a newly available Taq polymerase for longer product synthesis successfully amplified fragments representing BCL3/C alpha junctional sequences for t(14;19); c-MYC/C mu, c-MYC/C gamma and c-MYC/C alpha for t(8;14); BCL6/C lambda for t(3;22); BCL6/C kappa for t(2;3); 5'-BCL6/C mu and 5'-BCL6/C gamma for t(3;14), respectively. The sizes of the amplified fragments were varied from 1.8 kb to 12 kb, which were specific to each material. Present study provides a useful tool for diagnosis and subsequent management of B-cell lymphoma/leukemia characterized with specific chromosomal translocation. PMID- 9209374 TI - LIM-only protein Lmo2 forms a protein complex with erythroid transcription factor GATA-1. AB - The LIM-only protein Lmo2, originally identified as an oncogenic protein in human T cell leukemia, is essential for erythropoiesis. A possible role for Lmo2 in transcription during erythropoiesis has been investigated. Direct interaction of Lmo2 was observed in vitro and in vivo with the zinc finger transcription factor GATA-1, as well as with the basic helix-loop-helix (bHLH) transcription factor Tall. By using mammalian two-hybrid analysis, E47/Tall/Lmo2/GATA-1 protein complex could be demonstrated. Thus, a molecular link exists between three proteins crucial for erythropoiesis. This data suggest that variations in amounts of complexes involving Lmo2, Tall, and GATA-1 could be important for erythroid differentiation. PMID- 9209377 TI - Rearrangement of the BCL6 gene in B-cell lymphoid neoplasms. AB - We report here a large series of B-cell neoplasms with regard to rearrangement of the BCL6 gene on chromosome band 3q27. Southern blot analysis using probes from the major translocation cluster (MTC) region of the BCL6 revealed rearrangement in 32 of a total of 222 patients with various subtypes of B-cell neoplasm. In non Hodgkin's lymphoma (NHL), rearrangements of the BCL6 gene were not closely associated with a specific histopathologic subtype but distributed in subcategories in the Working Formulation. A comparative study between NHL associated either with BCL2 or BCL6 rearrangement showed that advanced disease and bone marrow involvement were more frequent in BCL2(+) NHL. In contrast, extranodal involvement was more frequently observed in the BCL6(+) NHL. The survival curve of BCL6(+) NHL was characterized by a rapid decline followed by a plateau. Of the total of 32 BCL6(+) patients, 6 carried both BCL2 and BCL6 rearrangements, and showed clinicopathological properties of follicular lymphoma. This study suggests that BCL6 rearrangement is primarily associated with large cell lymphoma, and that BCL2(-)BCL6(+) NHL could potentially be curable with modern combination chemotherapy. PMID- 9209378 TI - p53-induced p21 controls DNA replication. AB - The p21 protein, a regulator of cyclin-dependent kinases (CDKs), has been thought to be one of the key proteins to function in cell proliferation suppression upon DNA damage. In normal cells but not in many tumor cells, p21 forms a quaternary complex with a cyclin, a CDK and the proliferating cell nuclear antigen (PCNA), one of the DNA replication and repair factors, suggesting that this complex might play an important role in maintaining the integrity of the genome. Here, we have focused on the p21-PCNA interaction in the context of DNA replication or DNA repair, presenting the data from both in vitro and in vivo studies of the p21 function. PMID- 9209379 TI - p53-mediated cell cycle arrest and apoptosis. AB - We have generated a series of murine erythroleukemia clones that ectopically express a temperature sensitive mutant p53 allele. In many clones, activation of p53 at low temperature resulted in the accumulation of cells in G1 and in apoptosis. Several cytokines including erythropoietin, IL-3 and the ligand for the Kit receptor blocked p53-dependent apoptosis in p53ts-expressing cells at 32 degrees C. Cytokine-treated cells were reversibly arrested in G1 and resumed growth upon return to 37 degrees C. Certain clones exhibited only a G1 arrest in response to p53 activation at 32 degrees C. One of the these clones secreted erythropoietin and another secreted IL-3. We tested the possibility that autocrine secretion of IL-3 played a role in preventing apoptosis and showed that disruption of the autocrine loop by cell dilution or with neutralizing antibodies to IL-3 restored p53-dependent apoptosis at 32 degrees C. Thus, two properties of p53 protein, namely, its ability to arrest cells in G1 and its ability to promote apoptosis could be uncoupled by cytokines acting as survival factors. PMID- 9209380 TI - Expression of p53 in differentiation and apoptosis and its deregulation in tumor cell. AB - The observation that wild type p53 may induce cells to undergo either apoptosis or differentiation raises the question of whether these two events share similar p53-dependent pathways. To evaluate the interrelationship between these two p53 dependent processes, our study focused on the human HL-60, a pro-myelocytic p53 non-producer cell line in which p53 expression was introduced and the induction of apoptosis and differentiation was followed under controlled conditions. p53 expression was induced in the HL-60 cell line by infection with the recombinant wild type p53 (p53WT) vaccinia virus. Viral infection gave rise to cells expressing various levels of wild type p53 protein. High levels of p53 protein induced cells to undergo rapid apoptosis, whereas lower levels of p53 protein induced cells to undergo cell differentiation at a more moderate rate of kinetics. These results suggest that p53 protein levels may determine whether a given cell should prefer one pathway over the other to exit the cell cycle. PMID- 9209381 TI - Role of p53 tumor suppressor gene and Fas/Apo-1 in induction of apoptosis and differentiation of cancer cells. AB - Recent studies have suggested that wild-type p53 blocks cell cycle progression near the G1-S boundary and is involved in both differentiation and apoptosis in many types of cells including cancer cells. p53 expression is enhanced upon DNA damaging apoptotic stimuli while Fas/Apo-1, a member of the tumor necrosis factor receptor family expressed on cell surface, transduces a signal for apoptosis upon specific ligand or antibody engagement. We demonstrated that stable transfection of the wild-type p53 gene under the control of CMV promoter induced differentiation and apoptosis under restricted conditions in cancer cells, and often caused sensitization of p53-transfected cells to Fas/Apo-1 signal. To investigate the interaction between two major apoptotic pathways involving p53 and Fas/Apo-1 we have established a system that allows to induce wild-type p53 overexpression and apoptosis in cancer cells upon treatment with anti-Fas antibody. The system also allows to investigate other factors interacting with p53 and Fas/Apo-1, and should provide a clue to understanding the biological and biochemical aspects of apoptosis. PMID- 9209382 TI - Genetic instability of p53-deficient mouse cells. AB - We established fibroblast cultures from p53-deficient mouse embryos, which were originally constructed by Dr. S. Aizawa (Kumamoto Univ.). These p53-deficient fibroblasts showed the same sensitivity to UV and X-ray as wild-type fibroblasts. There was no difference between repair activity of UV-induced DNA damages in p53 deficient and wild-type cells, either. However, UV-induced sister chromatid exchanges were significantly increased and delay of entering S-phase after UV irradiation was reduced in p53-deficient cells indicating abnormality in the checkpoint function of the cell cycle in p53-deficient cells. We also used the supF gene on a shuttle vector to analyze UV-induced mutations in p53-deficient cells. Although frequencies of UV-induced mutations were not different between p53-deficient and wild-type cells, distributions of base-substitution mutations on the supF gene were different. PMID- 9209383 TI - p53 mediated apoptosis in HeLa cells: transcription dependent and independent mechanisms. AB - The most frequent target for genetic alterations in human cancers is the p53 tumor suppressor gene. Mutations in p53 abrogate its ability to inhibit cell growth and to suppress tumor progression. The anti-proliferative activity of p53 can be mediated by the induction of growth arrest and/or programmed cell death (apoptosis). Recent in vivo studies support the involvement of apoptosis in tumor suppression by p53. To gain further insight into the mechanisms by which p53 induces apoptosis, the activity of p53 was studied in HeLa cells using a transient transfection assay. To define the functional domains of p53 required for apoptosis a C-terminal deletion mutant of p53 was used. This mutant, p53d1214, lacks the oligomerization domain, the nuclear localization signal and a large part of the core DNA binding domain. This mutant was shown to be deficient in sequence specific transactivation activity. Overexpression of wt p53 induced an efficient apoptosis in transiently transfected HeLa cells. Surprisingly p53d1214, containing only the first 214 N-terminal residues induced extensive apoptosis. The induction of apoptosis by p53d1214 is slower than that induced by wt p53. Furthermore, p53d1214 suppressed the transformation of rat embryo fibroblasts by several oncogene combinations, such as myc plus ras. In view of the fact that p53d1214 lacks measurable transactivation potential, our findings suggest the existence of two p53 dependent-apoptotic pathways--one involves activation of specific target genes, and the other is independent of it. Transactivation independent apoptosis may play a central role in tumor suppression by p53. PMID- 9209384 TI - Possible involvement of MSX-2 homeoprotein in v-ras-induced transformation. AB - A truncated MSX-2 homeoprotein was found to induce flat reversion when expressed in v-Ki-ras-transformed NIH3T3 cells. Although the expression of endogenous MSX-2 gene is low in most of the normal adult tissues examined, it is frequently activated in carcinoma-derived cell lines. Likewise, the gene is inactive in untransformed cells but is transcriptionally activated after transformation by v Ki-ras oncogene, suggesting that the intact MSX-2 may play a positive, rather than suppressive, role in cell transformation. To test this possibility, we isolated a full-length human MSX-2 cDNA and tested its activities in two cell systems: fibroblast and myoblast. In NIH3T3 fibroblasts, although the gene by itself failed to confer a transformed phenotype, antisense MSX-2 cDNA as well as truncated MSX-2 cDNA interfered with the transforming activities of both v-Ki-ras and v-raf oncogene. In C2C12 myoblasts, MSX-2 was found to suppress MyoD gene expression, as do activated ras oncogenes, under certain culture conditions, and truncated MSX-2 cDNA was found to inhibit the activities of both MSX-2 and ras in this system as well. Our findings not only suggest that the truncated version MSX 2 may act as a dominant suppressor of intact MSX-2 but also raise the possibility that MSX-2 gene may be an important downstream target for the Ras signaling pathways. PMID- 9209385 TI - Point mutation of p53 tumor suppressor gene in bovine leukemia virus-induced lymphosarcoma. AB - To elucidate the mechanism of leukemogenesis induced by bovine leukemia virus (BLV), the abnormality of p53 tumor suppressor gene was examined using the sequencing method of polymerase chain reaction (PCR)-amplified DNA from peripheral blood lymphocytes (PBL) and tumors from BLV-infected cattle that showed evidence of different stages during the progression of enzootic bovine leukosis (EBL). Mutations of the p53 gene were found in tumor cells from cattle with EBL, but not in PBL from BLV-free normal cattle or BLV-infected cattle without any evidence of tumor, suggesting mutation of p53 gene occurred specifically at the lymphoma stage of the disease. Twelve of eighteen cattle with EBL had seven missense and five silent mutations. The mutations were mapped to the highly evolutionarily conserved regions of p53 gene, and were involved in the DNA binding of p53. Thus, it appeared that the p53 point mutation is one of the critical events leading from the asymptomatic stage to the lymphoma stage. PMID- 9209386 TI - Control of G1 progression by D-type cyclins: key event for cell proliferation. AB - Mammalian D-type cyclins are differentially expressed during the first gap phase (G1) of the cell cycle in various cell types, and function as regulatory subunits of cyclin-dependent kinases (cdks), cdk4 and cdk6, to form holoenzymes whose activities are both necessary and rate limiting for G1 progression. Mitogenic signals induce the expression of cyclin D and cdk4 proteins, and facilitate their assembly into holoenzymes and their post-translational modification, while anti proliferative stimuli extinguish the activity of cyclin D-dependent kinases by inducing cdk inhibitors which directly interfere with their catalytic functions and/or inhibit the post-translational activation of cyclin-bound cdks. Therefore, a variety of extracellular signals target and regulate the cyclin D/cdk4 serine/threonine kinases, which execute their critical functions during middle to late G1 phase by phosphorylating key substrates, including the retinoblastoma tumor suppressor gene products (pRb). Although overexpression of cyclin D, or inactivation of Rb or cdk inhibitor gene alone is not sufficient for cell transformation, high frequency of alterations of these genes in cancers suggests that inactivation of this particular pathway is involved in tumor development. PMID- 9209387 TI - Regulation and function of the cyclin-dependent kinase inhibitor p16CDKN2. AB - Many human tumours show perturbations of a pathway that includes the D-cyclins, their associated cyclin-dependent kinases, and specific kinase inhibitors. The focal point of this pathway is the product of the retinoblastoma tumour suppressor gene, pRb, which imposes a block on G1 phase progression. Thus, the major role of the cyclin D-dependent kinases is to overcome this block by initiating the phosphorylation of pRb. Excessive activity of this pathway is likely to lead to excessive cell proliferation. Conversely, accumulation of the inhibitors is associated with the cessation of cell division. PMID- 9209388 TI - Two different bindings of p21 Cdk inhibitor to cyclin/Cdk complex. AB - The cyclin-dependent kinase inhibitor p21(p21 CKI) has been found to inhibit the activity of several Cdks. Of interest wre reports that more than one molecule of p21 CKI appear to be necessary for Cdk kinase inhibition. In this report, we first determined the two different regions of p21CKI that were important for binding to cyclin D1/Cdk4 complex. Mutant analysis revealed that the either binding site is enough for their binding but for Cdk4 kinase inhibition both sites are necessary. Since the mutants (delta 17-22 or W49G) which lacks either binding site could complement each other for kinase inhibition, two molecules of p21 CKI with different binding mode might be a mechanism of cyclin D1/Cdk4 kinase inhibition. PMID- 9209389 TI - Recent progress in molecular mechanisms of leukemogenesis: the cyclin-dependent kinase 4-inhibitor gene in human leukemias. AB - In order to clarify the significance of p16 gene (CDKN2) inactivation and its disease specificity among hematopoietic tumors, configurations of the p16 gene as well as those of the adjacent p15 and interferon alpha (IFN alpha) genes were examined in primary hematopoietic tumors. Loss of the p16 gene is frequent in and highly specific to lymphoid tumors among hematopoietic tumors. Gene deletions but not minute mutations should be the predominant mechanism of p16 gene inactivation in these types of tumors. The p16 gene is most frequently deleted among the p16, p15 and IFN alpha genes and thus should be the target of deletions in this locus. Deletions of the p16 gene were frequently observed in tumors carrying chromosome 9p abnormalities while a significant number of cases showed loss of the p16 gene without chromosome 9p abnormalities. So far inactivation of p53 and Rb tumor suppressors have also been found in lymphoid tumors. In our study, we detected homozygous deletions of p16 gene in 20%, loss of Rb protein in 28%, and p53 gene alterations in 8% of lymphoid tumors. Notably, 44% of lymphoid tumors showed inactivation of at least one of the three tumor suppressors, suggesting these tumor suppressors are important for lymphoid tumorigenesis. Inactivations of these tumor suppressors should independently occur in development of lymphoid tumors. PMID- 9209390 TI - Contact inhibition-induced inactivation of the cyclin D-dependent kinase in rat fibroblast cell line, 3Y1. AB - Inhibitory proteins for Cdks (CKIs) are involved in cell cycle arrest induced by anti-mitotic factors, chemicals, or DNA damage in mammalian cells. High cell density also induces cell cycle arrest with unreplicated genomic DNA even in the presence of mitotic dose of the growth factors, termed contact inhibition. Although rat fibroblast cell line, 3Y1, arrested in quiescence by contact inhibition, Cdk4 bound its regulatory subunit, cyclin D1 or D3. However, these complexes were enzymatically inactive. Phosphorylation of the cyclin D1-bound Cdk4 by Cdk4-activating kinase, composed of cyclin H and MO15 (alias Cdk7), which was reconstituted in Spodoptera frugiperda cells (Sf9) could convert inactive cyclin D1-Cdk4 complex into active form in vitro, suggesting that threonine 172 in the Cdk4, whose phosphorylation is required for its activation, was in part unphosphorylated in the contact-inhibited 3Y1. Although MO15 was active in cell extracts prepared from the contact-inhibited 3Y1, activation of bacterially produced Cdk4 in the cell extracts was inhibited. Removing p27kip1 from the cell extracts allowed MO15 holoenzyme to phosphorylate the Cdk4 and to activate it, indicating that an access of MO15 to Cdk4 was inhibited by p27kip1 in the contact inhibited 3Y1. PMID- 9209391 TI - Kip1 degradation via the ubiquitin-proteasome pathway. AB - The cell cycle has been the object of extensive studies for the past years. A complex network of molecular interactions has been identified. In particular, a class of cell cycle inhibitory proteins has been identified but details of the molecular mechanism of their action have yet to be resolved. These inhibitors regulate the progression through G1 and the G1/S transition via the inhibition of the cyclin-dependent kinase (Cdk) activity. The potential function of these negative regulators as tumor suppressors provides new insights into the link between the cell cycle and oncogenesis. Kip1 is a potent inhibitor of Cdks. In quiescent cells Kip1 accumulates without an increase in mRNA or protein synthesis. We demonstrated that cell cycle regulation of Kip1 levels, both in normal and transformed human cells, occurs via the ubiquitin-proteasome pathway. In a crude in vitro system, Kip1 is ubiquitinated and degraded in an ATP dependent manner and inhibition or depletion of the proteasome blocks Kip1 degradation. Human Ubc2 and Ubc3, the homologs of yeast Rad6 and Cdc34 gene products respectively, are specifically involved in the ubiquitination of Kip1. Compared to proliferating cells, quiescent cells contain a far lower amount of Kip1 ubiquitinating activity. These results represent the first demonstration that the ubiquitin-proteasome pathway plays a role in the regulation of a cell cycle protein in human cells, namely the Cdk inhibitor Kip1. The specific proteolysis of Kip1 may be involved in the pathway of inactivation of Cdks. PMID- 9209392 TI - Involvement of the cyclin-dependent kinase inhibitor p27Kip1 in negative signaling through the antigen-receptor of B lymphocytes. AB - Several lines of evidence suggest that interaction with antigens generates a negative signal via the antigen receptor of B lymphocytes (cell surface immunoglobulin; sIg), resulting in apoptosis, growth arrest or functional inactivation, and that activation of B cells requires an additional co stimulatory signal such as a T cell-derived signal through the B cell membrane molecule CD40. In the B cell line WEHI-231, sIg crosslinking induces apoptosis and cell cycle arrest at the late G1 phase, both of which are reversed by CD40 signaling. Crosslinking of sIg reduces the activity of cyclin dependent kinase (Cdk)2 required for cell cycle progression in the late G1 phase by induction of a Cdk inhibitor (CKI) p27Kip1, but the induction of p27Kip1 is abrogated by CD40 signaling. These results strongly suggest that p27Kip1 plays some role in negative signaling via sIg, resulting in growth arrest of antigen-stimulated B cells. PMID- 9209393 TI - Cyclin-dependent kinase inhibitors in human neoplasms. PMID- 9209395 TI - Retinoid receptors in development and disease. AB - Nuclear receptors comprise a large family of ligand-dependent transcription factors that display considerable specificity in and selectivity in regulating the genetic programs they ultimately influence. The response to retinoic acid (RA) is mediated by two families of transcription factors which include the retinoic acids receptors (RARs) and the retinoid X receptors (RXRs). In human acute promyelocytic leukemia (APL), RAR alpha becomes an activated oncogene as a consequence of its fusion to the PML locus. Because patients with APL can be induced into remission with high dose RA therapy, we propose that PML-RAR is a new class of dominant negative oncogene that disrupts a structure that includes at least five other proteins. This mega-complex, referred to as a "POD", is disrupted in leukemic cells expressing the oncoprotein and is reassembled following high dose RA therapy in both cell culture an in patients. PMID- 9209394 TI - Cloning of feline p21WAF1 and p27Kip1 cDNAs and search for their aberration in leukemias and lymphomas in cats. AB - For investigation of the relation of cell cycle regulation with tumorigenesis in cats, we cloned feline p21WAF1 and p27Kip1 cDNAs and searched for their aberration in feline spontaneous leukemias and lymphomas. The feline p21WAF1 cDNA (pCFW.31) clone obtained from the PCR amplified product appeared to cover approximately 75% of the open reading frame, and showed 81.6% and 76.8% sequence similarities with those of human and mouse counterparts, respectively. The pHFK.5 clone isolated by plaque hybridization contained the whole open reading frame of cat p27Kip1 cDNA encoding 198 amino acids, showing 93.4% and 90.4% sequence similarities with those of human and mouse counterparts, respectively. Southern blot analyses using these clones as probes did not show any deletion or rearrangement of both the p21WAF1 and p27Kip1 genes in 19 feline spontaneous cases of leukemias and lymphomas examined. RT-PCR/SSCP (single strand conformation polymorphism) analysis of p27Kip1 cDNA indicated that there was no mutation resulting in amino-acid substitution in 10 feline leukemia and lymphoma cases. PMID- 9209396 TI - Unravelling of physiological functions of retinoic acid using a dominant-negative retinoic acid receptor. AB - We have constructed dominant-negative retinoic acid receptors (RARs) by substituting single amino acid which has been found in a dominant-negative thyroid hormone receptor, and have expressed the dominant-negative RAR in the epidermis, a potential target organ of retinoic acid (RA). The resultant transgenic mice exhibited dramatic suppression of epidermal development, demonstrating the absolute requirement of RA in normal skin development. This method is theoretically applicable to every organ, thus opening the way to define the physiological roles of RA during embryogenesis as well as in adults. PMID- 9209397 TI - Bcl-2 and Bcl-xL block apoptosis as well as necrosis: possible involvement of common mediators in apoptotic and necrotic signal transduction pathways. AB - The proto-oncogene bcl-2 and a bcl-2-related gene bcl-x prevent apoptotic cell death induced by various treatments. Although a mechanism has been proposed that involves Bcl-2 activity on reactive oxygen species (ROS), we find that expression of Bcl-2 or Bcl-xL prevents cell death induced by withdrawal of oxygen (hypoxia) and that the cell death does not involve ROS, suggesting that Bcl-2 or Bcl-xL exerts an anti-cell death function by a mechanism other than through regulation of ROS activity. Using electron microscopy, and confocal and non-confocal fluorescence microscopy, we show that hypoxia induces both necrosis and apoptosis. Overexpression of Bcl-2 or Bcl-xL blocks hypoxia-induced apoptosis and, although to a lesser extent, necrosis. The anti-apoptotic proteins Bcl-2 and Bcl-xL effectively inhibit KCN-induced cell death which is characterized by necrotic features including apparently intact chromatin, remarkable mitochondrial swelling with loss of crista structure and loss of plasma membrane integrity. The necrotic cell death is also inhibited by inhibitors of ICE (interleukin-1 beta converting enzyme)(-like) proteases, the common mediators of apoptosis. These results indicate that Bcl-2/Bcl-xL and ICE(-like) proteases modulate both apoptotic and at least some forms of necrotic cell death, suggesting that both cell death pathways involve some common mediators. PMID- 9209398 TI - Deregulation of cell survival in lymphomagenesis. AB - Because normal lymphoid tissue homeostasis depends on a balance between cell proliferation and cell death, lymphomas can arise from mutations that interfere with the normal cell death process. We here discuss some circumstances in which lymphoid cell overexpression of a cell death antagonist, the Bcl2 protein, in E mu-bcl2 transgenic mice can contribute to the development of lymphomas and plasmacytomas. PMID- 9209399 TI - Characterization of a mammalian cell death gene Nedd2. AB - Through subtraction cloning approach, we have identified a set of mouse genes with developmentally down-regulated expression in brain. One such gene, termed Nedd2, was found to encode protein similar to the mammalian interleukin-1b converting enzyme (ICE) and the product of the nematode (C. elegans) cell death gene ced-3. Our data suggest that Nedd2 is an important component of the mammalian programmed cell death machinery. PMID- 9209400 TI - Granulocyte-colony stimulating factor induced apoptosis in radiation-induced murine leukemia cell line. AB - Cytokines regulate proliferation and differentiation of hematopoietic progenitor cells. Recently it has been clarified that physiological cell death, apoptosis plays important role of hematopoiesis. So we evaluated the effects of granulocyte colony stimulating factor (G-CSF) on leukemic cells, especially focused on apoptosis. Intravenous inoculation of radiation-induced murine leukemia cell line, C2M-A5 into the parent C3H mice resulted in the development of myeloid leukemia. However, the leukemic death of the mice was completely suppressed by the daily subcutaneous injection of recombinant human (rh)G-CSF from the next day (Bessho M. et al., Leuk Res 1989:13:1001-1007). In the in vitro study using C2M A5 cells, we found that apoptosis appears on the cells at 48 hours after addition of G-CSF in culture. The cells in this stage lost the leukemogenicity to C3H mice (Bessho M. et al., Leukemia 8:1185-1190:1994). To clarify the mechanism of the induction of apoptosis by G-CSF we studied cell cycle and molecular changes in C2M-A5 cells cultured in medium with or without rhG-CSF by means of using the flowcytometry and Northern and Western blot analyses. After addition of rh G-CSF to culture, C2M-A5 cells removed to S phase, next arrested at G0/G1 phase on and after 24 hours, and 48 hours later, apoptosis was observed. Overexpression of mRNAs for c-myc (3-24 hours later) and for p53 (6-24 hours later), were observed in the cell cultured in rhG-CSF administered medium with a concomitant down expression of bcl-2 mRNA (from 6 hours later). Tyrosine-phosphorylated protein (17 kd) appeared at 48 hours after administration of rhG-CSF to cell culture. This protein was suggested for specific apoptosis induction by rhG-CSF. These results are summarized as follows. (1) rhG-CSF induced apoptosis to C2M-A5 and deprived its leukemogenicity to mice. (2) Induction of apoptosis was associated with cell cycle and correlated to the changes of the expression rates of c myc,p53, and bcl-2. (3) Tyrosine kinase may play an important role in apoptosis induction to C2M-A5 by rhG-CSF. PMID- 9209401 TI - Up-regulation of cell cycle-associated genes by p53 in apoptosis of an erythroleukemic cell line. AB - A murine erythroleukemic cell line (1-2-3) which expresses only the temperature sensitive mutant p53 gene (Ala-to-Val substitution at codon 135) was established. When these cells were cultured at 32 degrees C, the growth rate was reduced significantly and DNA fragmentation, a typical character of apoptosis, was observed. In this process, p53 migrated from cytoplasm to nucleus and protein complexes binding to the p53-responsive element were detected in nuclear extracts of the cells cultured at 32 degrees C by gel-shift assay and transactivation from the p53-responsive element was detected. The expression of the p21 (waf1/cip1/sdi1), cyclin G and gadd45 genes was increased (about 3 to 4 fold that at 37 degrees C), when the cells were cultured at 32 degrees C. However, the expression of the bax gene was increased slightly (about 1.5 fold that at 37 degrees C) and no significant change was detected in expression of the mdm2 gene. No change in the amount of Fas antigen was observed by flow cytometric analysis. Transcripts of the bcl-2 and fasl gene were not detected in the cells both at 37 degrees C and 32 degrees C. These results suggest that up-regulation of the genes associated with the cell cycle and/or DNA replication, such as p21, cyclinG and gadd45 rather than bax, fas, fasl and bcl-2 may be important for induction of apoptosis of this erythroleukemic cell line by p53. PMID- 9209402 TI - The specific N-ras mutation in rat 7,12-dimethylbenz[a]anthracene (DMBA)-induced leukemia. AB - Intravenous injections of 7,12-dimethylbenz[a]anthracene (DMBA) induce erythroblastic leukemia (erythroleukemia) with #2 trisomy and Long #2 in Long Evans rats. Recently, a consistent type of mutation, A to T transversion in codon 61 of N-ras gene, was found in all of 6 cultured leukemia cell lines and 13 primary leukemias induced by DMBA using polymerase chain reaction (PCR) and direct sequencing. On the contrary, no mutation was observed in Ha- and Ki-ras genes in these leukemias. The consistent occurrence of the above N-ras mutation in DMBA-induced leukemias indicates that N-ras gene plays an important role in DMBA-leukemogenesis. Mutations in ras genes generally takes place during the initiation stage of carcinogenesis because they often appear in the premalignant stage of tumors. In order to detect the N-ras mutation in an early stage of leukemogenesis, we designed the mutant-allele-specific amplification (MASA) method to detect the mutation in bone marrow (BM) cells of DMBA-treated rats. The MASA method was sensitive enough to detect one mutant cell mixed in 10(6) normal cells. Using this method, the N-ras mutation was found in BM cells 2 days after single DMBA injection and thereafter throughout the preleukemic stage. These results suggest that the N-ras mutation is an earliest event in DMBA-induced leukemogenesis. PMID- 9209403 TI - Mechanisms of constitutive activation of c-kit receptor tyrosine kinase. AB - We investigated the mechanism of constitutive activation of c-kit receptor tyrosine kinase (KIT) found in the FMA3 murine mastocytoma cell line, and compared it with the mechanisms observed in other tumor mast cell lines (the HMC 1 human mast cell leukemia cell line, the RBL-2H3 rat mast cell leukemia cell line, and the P-815 murine mastocytoma cell line). The c-kit gene obtained from FMA3 cells was found to have 21-base deletion at the juxtamembrane domain of KIT, thereby leading to the constitutive activation of KIT. The deletion at the juxtamembrane domain resulted in constitutive dimerization of c-kit proteins, whereas the point mutation that were detected at the kinase domain of KIT in HMC 1, RBL-2H3, and P-815 cells caused constitutive activation of KIT without dimerization. These constitutively activating mutations of c-kit may play a role in development of mast cell tumors. PMID- 9209404 TI - Isolation and analysis of cellular DNA fragments directly binding to c-Myc protein. AB - c-Myc protein, the product of cellular oncogene c-myc, is thought to play an important role in the control of cell cycle progression by binding to the E-box sequence (CACGTG) of cellular DNA, but only a few target genes are known. We cloned two small human DNA fragments (n16 and r37) that bound to c-Myc protein in vitro by random screening. Both clones contained the E-box sequence, to which c Myc protein bound directly in vitro. Northern blot analysis showed that a low molecular-weight RNA was transcribed from the region near the n16 c-Myc binding site. The function of this low molecular-weight RNA and the regulatory role of c Myc protein in related transcription are now under investigation. PMID- 9209405 TI - v-Rel activates the proto-oncogene c-Jun promoter: a correlation with its transforming activity. AB - v-Rel is a transforming protein of the reticuloendotheliosis virus, and is a transcription factor regulating various cellular genes. We found that v-Rel activates the promoter of the proto-oncogene c-jun in a transient transfection assay system. Moreover, the expression of endogenous c-jun was augmented in cells expressing exogenous v-Rel, but not c-Rel. The transcriptional activities of v Rel to the tested promoters containing the kB-site are lower than that of c-Rel, but that to the c-jun promoter was much higher than that of c-Rel. The N-terminal DNA binding domain of v-Rel, which is responsible for its high transforming activity of v-Rel was also responsible for the high transcriptional activity to the c-jun promoter. Thus, the activity of v-Rel upon the c-jun promoter correlates well with its transforming ability. Since c-Jun plays pivotal roles on cell proliferation in various types of cells, the activation of c-jun expression by v-Rel may be an essential step for the oncogenic transformation caused by v Rel. PMID- 9209406 TI - The signal transduction through Grb2/Ash in hematopoietic cells. AB - Grb2/Ash is composed of one SH2 and two SH3 domains and functions as an adapter linking tyrosine-kinase receptors and Ras in fibroblasts. The SH2 domain binds to tyrosine-phosphorylated proteins and the SH3 domain binds to protein containing proline-rich regions. However, the mechanisms of signal transduction through Grb2/Ash in hematopoietic cells are still unclear. By means of the binding experiments using the GST fusion protein including the full length Grb2/Ash, we have found that Shc and unidentified 130-kDa and 135-kDa proteins are associated with Grb2/Ash and that they are tyrosine-phosphorylated by treatment with granulocyte-macrophage colony-stimulating factor (GM-CSF) and erythropoietin (EPO) in a human leukemia cell line UT-7. We have purified the 130-kDa protein (pp 130) using GST-GRB2/Ash affinity column. The amino-acid sequence analysis showed that the pp130 was identical to the human c-cbl proto-oncogene product (c Cbl). c-Cbl constitutively binds to the SH3 domain of Grb2/Ash both in vitro and in vivo but not to the SH2 domain of Grb2/Ash. Moreover, c-Cbl (pp 130) becomes tyrosine-phosphorylated rapidly and transiently depending on GM-CSF and EPO stimulation. However, we could not find the homologous regions with guanine nucleotide exchange factors or GTPase-activating proteins in the c-cbl gene. These findings strongly suggest that c-Cbl is implicated in the signal transduction of GM-CSF and EPO in hematopoietic cells, and c-Cbl and Grb2/Ash might also transduce a signal that is different from the signal leading to Ras regulation. Recently, we have shown that the proto-oncogene vav product (Vav) is also tyrosine-phosphorylated by treatment with GM-CSF and EPO and is constitutively associated with the SH3 domain of Grb2/Ash in UT-7. Another guanine nucleotide exchange factor Sos is also associated with Grb2/Ash in UT-7. It has been reported that Vav has guanine nucleotide exchange activity and activates Ras in vitro and in vivo. These data suggest that tyrosine kinases, the adapter Grb2/Ash, and the guanine nucleotide exchange factor Vav and Sos are members of a signaling pathway leading to Ras activation in hematopoietic cells. PMID- 9209407 TI - Chemical modification of L-asparaginase with comb-shaped copolymer of poly(ethylene glycol) derivative and maleic anhydride. AB - L-asparaginase from Escherichia coli, an antitumor enzyme, was chemically modified with a comb-shaped copolymer of poly(ethylene glycol) derivative and maleic anhydride (activated PM). The PM-modified asparaginase lost the immunoreactivity with retaining high enzymic activity and also prolonged the clearance time in blood. Intraperitoneal administration of PM-asparaginase markedly increased the mean survival-time of lymphoma L5178Y-bearing mice in comparison with that of unmodified asparaginase. Pretreatment of mice with PM asparaginase before immunizing with unmodified asparaginase extremely suppressed the anti-asparaginase antibody production. PMID- 9209409 TI - Cytokine regulation: an overview. PMID- 9209408 TI - Radiation-induced myeloid leukemia in mice under calorie restriction. AB - The host-defense mechanisms against cancers are known to be modulated by changing the environmental factor(s). The spontaneous incidence of myeloid leukemia is about 1% in C3H/He mice, and the incidence increases up to 23.3% when a single dose of radiation, 3 Gy X-ray, is exposed to a whole-body. Since calorie restriction was known to reduce the incidence of spontaneous tumors, a question as to whether such radiation induced-increase of myeloid leukemia would be also decreased by calorie restriction, was aimed to answer to elucidate possible mechanism of radiation-induced myeloid leukemia. By the calorie restriction, the incidence of myeloid leukemia was significantly decreased. In addition, the latent period of the myeloid leukemia in the groups for calorie restriction was significantly extended at a greater extent as compared with the control diet groups. Number of hemopoietic stem cells, the possible target cells for radiation induced leukemias, in the groups for the calorie restriction demonstrated a significant decrease, especially in the spleen, as compared with that in the control, when the evaluation was made at the time of radiation exposure. PMID- 9209410 TI - Regulation of IL-2 signaling. AB - Several tyrosine kinases such as Jak1, Jak3, Lck and Syk are known to participate in IL-2-mediated intracellular signal transduction. Jak1, Lck and Syk are associated with the cytoplasmic domain of the beta chain, whereas Jak3 is associated with the cytoplasmic domain of the gamma chain, which is shared among receptors for IL-2, IL-4, IL-7 and IL-15. We first demonstrated that Jak1 is associated with the alpha chains of receptors for IL-4, IL-7 and IL-15 as well as the IL-2 receptor beta chain. Furthermore, we revealed that two proline residues in the box1 region, which is conserved in the IL-2 receptor beta chain and the alpha chains of the cytokine receptors, are essentially involved in association with Jak1. The MOLT4 transfectants with the box1 mutants of the IL-2 receptor beta chain lacking Jak1 association showed IL-2 responsiveness, in terms of activations of Jak3 and Stat5 and induction of cell growth, indicating that Jak1 is dispensable for IL-2-mediated cell growth signaling, and that Jak1 activation is not required for activation of Jak3 and Stat5 in the MOLT4 transfectants. PMID- 9209411 TI - Signal transduction by the GM-CSF, IL-3 and IL-5 receptors. PMID- 9209412 TI - The role of erythropoietin receptor tyrosine phosphorylation in erythropoietin induced proliferation. AB - Although studies with truncated erythropoietin receptors (EpoRs) have suggested the tyrosine phosphorylation (Yphos) of the EpoR may not play a significant role in Epo-induced proliferation, we found, using a full length EpoR mutant designed Null, in which all 8 of the intracellular tyrosines (Ys) were substituted with phenylalanines (Fs), that Null cells required 5-10 fold more Epo than wild type (WT) EpoR containing cells in order to proliferate as well. Moreover, a comparison of Epo-induced proliferation with Epo-induced Yphos patterns, using DA 3 cells expressing WT, Null and various Y to F EpoR point mutants revealed that Stat5 Yphos and activation correlated directly with proliferation and was mediated primarily throuhg the most membrane proximal Y, i.e., Y343, although other tyrosines (most likely Y401 and Y431) within the EpoR could activate Stat5 in its absence. We also found that EpoR Yphos was essential for the Yphos of Shc and for the Yphos and association of a 145 kDa protein with Shc. We purified and cloned this Shc-associated 145 kDa protein and found that it was a unique SH2 containing inositol polyphosphate-5-phosphatase. This novel enzyme, which we have called SHIP for SH2-containing inositol-phosphatase, may modulate both Ras and inositol signaling pathways. PMID- 9209413 TI - Thrombopoietin: basic biology and clinical promise. PMID- 9209414 TI - P210 Bcr-Abl interacts with the interleukin-3 beta c subunit and constitutively activates Jak2. AB - Chronic myelogenous leukemia is a neoplasm of pluripotent hematopoietic cells. Cytokines such as interleukin-3 and granulocyte-macrophage colony-stimulating factor regulate the growth and differentiation of hematopoietic precursors. These cytokines activate two distinct signals to the nucleus. One signal is through the Ras pathway, and the second involves activation of Jak2. We demonstrated that Bcr Abl co-immunoprecipitates with and constitutively phosphorylates the common beta c chain of the interleukin-3 (IL-3) and granulocyte-macrophage-macrophage colony stimulating factor (GM-CSF) receptors. Our data show that formation of this complex leads to the constitutive activation of Jak2. Previously, it has been demonstrated that Bcr-Abl interacts with Grb2 and Shc, which in turn activates the Ras pathway. Thus, Bcr-Abl can activate signalling through both pathways in a factor-independent fashion. PMID- 9209415 TI - Activation of the JAK1-STAT5 pathway by binding of the Friend virus gp55 glycoprotein to the erythropoietin receptor. AB - Friend spleen focus forming-virus (F-SFFV) induces acute erythroleukemia in susceptible mice. Initiation of the erythroleukemia is due to binding of the env related glycoprotein gp55 encoded by F-SFFV to the erythropoietin receptor (EPOR). The gp55/EPOR interaction induces prolonged and growth factor independent proliferation in a factor-dependent cell line. In erythropoietin (EPO) signaling, the JAK2/STAT5 pathway was shown to be activated downstream of the EPOR to transmit the signal to the cells. To determine members of the JAK family and the STAT transcription factor family involved in the gp55/EPOR signaling, we examined tyrosine phosphorylation of JAKs and STATs in F-SFFV-infected erythroid or erythroleukemic cells. JAK1 and STAT5 were constitutively tyrosine-phosphorylated but the DNA binding activity of STAT5 was not induced without EPO stimulation in erythroblastoid cells from spleens of F-SFFV-infected mice and erythroleukemia cell lines derived from gp55-transgenic mice. These results indicate that JAK1 is involved in the gp55/EPOR signaling but STAT5 is not playing an essential role in the growth of those erythroid cells. PMID- 9209416 TI - Regulation of megakaryocytopoiesis by thrombopoietin and stromal cells. AB - Thrombopoietin (Tpo) is a cytokine which stimulates megakaryocyte maturation. We found that Tpo is constitutively and ubiquitously expressed in all tissues examined, including bone marrow stromal cells, even in thrombocytopenia, thrombosis and steady-state condition in mice. Thus, platelet level in circulation is not regulated by Tpo gene expression. Furthermore, when the purified megakaryocytes were cocultured with the stromal cells, most of the megakaryocytes adhered to the stromal cells and remained unchanged, while free megakaryocytes induced proplatelet formation. Thus the stromal cells in bone marrow secrete Tpo and stimulate megakaryocytopoiesis, but the interaction of megakaryocytes with the stromal cells may suppress platelet formation. Study on signal transduction through Mp1 revealed that Tpo induces activation of JAK2 and Tyk2, which in turn activate STAT1, STAT3 and STAT5. Further, Tpo stimulates transcription factors GATA-1 and NF-E2, which induce differentiation markers, GPIIb/IIIa and Pm-1. In addition, Shc, Vav, Ras, Raf-1, MAPKK, MAPK and Pim-1 are also activated. Thus, Tpo activates a lineage-specific cascade as well as a specific JAK-STAT cascade and a common signaling cascade. PMID- 9209417 TI - DNA damage-induced apoptosis and Ice gene induction in mitogenically activated T lymphocytes require IRF-1. AB - Lymphocytes are highly sensitive to DNA damage-induced apoptosis. In thymocytes, the tumor suppressor p53 has been shown to be required for this type of apoptosis. However an as yet unknown, p53-independent pathway(s) appears to mediate the same event in mitogenically activated mature T lymphocytes. By using mice with a null mutation in the IRF-1 gene, we revealed that DNA damage-induced apoptosis in the latter cell type is dependent on the anti-oncogenic transcription factor interferon regulatory factor-1 (IRF-1). Thus two different anti-oncogenic transcription factors, p53 and IRF-1, are required for distinct apoptotic pathways in T lymphocytes. Furthermore, we found that mitogen induction of the interleukin-1 beta-converting enzyme (Ice) gene, a mammalian homolog of the Caenorhabditis elegans cell death gene ced-3, is also IRF-1-dependent. An IRF 1 binding sequence was identified in the 5' flanking region of the Ice gene. In addition, ectopic overexpression of IRF-1 results in the activation of the endogenous Ice gene and enhances the sensitivity of cells to radiation-induced apoptosis. Thus, induction of Ice gene may be involved in IRF-1 dependent DNA damage-induced apoptosis in activated mature T lymphocytes. PMID- 9209419 TI - Role of alternative splicing of the rat erythropoietin receptor gene in normal and erythroleukemia cells. AB - An alternatively splicing of the rat erythropoietin receptor (EpoR) gene was identified in normal and erythroleukemia cells by the reverse transcription PCR method. Insertion of a 105 bp fragment at the region corresponding to the extracellular domain of rat EpoR was found. The insert sequence, which encodes additional 21 amino acids, is similar to that previously found in the mouse EpoR gene, however, has an additional 27 bp direct repeat. Due to the presence of a stop codon in the insert, the alternative transcript is translated to a truncated and soluble form of EpoR which is preferentially expressed in liver, spleen, kidney, heart, and bone marrow cells as well as cultured erythroleukemia cells. These findings suggest that alternative splicing of the EpoR gene may play an important role in proliferation and differentiation of rat erythroid cells. PMID- 9209418 TI - Prognostic significance of TNF alpha and its p55 soluble receptor in malignant lymphomas. AB - In 93 newly diagnosed lymphoma patients, tumor necrosis factor alpha (TNF alpha) and its p55 soluble receptor (p55-sR), were prospectively determined in plasma by IRMA and ELISA methods respectively. These 93 patients included 31 patients with low grade lymphoma, 55 with intermediate or high grade lymphoma and 7 with Hodgkin's disease. Median TNF alpha plasma values were 20 pg/mL (range 5-380 pg/mL) in patients versus 7 pg/mL (range 4-9 pg/mL) in healthy control subjects. Presence of TNF alpha level equal or greater than 20 pg/mL was significantly associated with elevated LDH level, serum beta 2-microglobulin level > or = 3 mg/L, hemoglobin < or = 12 g/dL, Ann Arbor stage III or IV disease, and with bulky tumor. High level of TNF alpha was also associated, although less strongly, with B symptoms, poor performance status, and serum albumin < or = 35 g/L, yet it was not associated with change in acute phase protein levels. Levels of p55-sR were also markedly elevated in these lymphoma patients (median of 3.5 ng/mL, range 0.8-18.8 ng/mL) versus 1.45 ng/mL in control subjects (range 1.1-2.3 ng/mL). Level of p55-sR equal or greater than 3.5 ng/mL was significantly associated with poor performance status, B symptoms, beta 2-microglobulin levels > or = 3 mg/L, serum albumin < or = 35 g/L, C-reactive protein > 6 mg/L, hemoglobin < or = 12 g/dL, and bulky tumor. In the whole group of 93 patients, both high TNF alpha and p55-sR levels strongly predicted short freedom from progression and overall survival. This study suggests that elevated TNF alpha and p55-sR plasma levels have a high correlation with other adverse prognostic factors in lymphoma patients and predict their poor outcome. PMID- 9209420 TI - Antileukemic effect of interferon-alpha is mediated through down-modulation of E2F activity. PMID- 9209421 TI - Cloning and biological activity of murine oncostatin M. AB - Oncostatin M (OSM) is a member of the interleukin-6/leukemia inhibitory factor (LIF) family cytokines. While human OSM (hOSM) has been characterized, the murine counterpart had not been isolated. We cloned a murine OSM (mOSM) cDNA as a gene that is induced in hematopoietic cells by a subset of cytokines including IL-3, GM-CSF and Epo. Identity of mOSM was based on overall homology to hOSM and chromosomal gene localization. Human OSM is known to exhibit biological activities similar to LIF, because they share the same functional receptor composed of the LIF receptor and gp130. As compared to hOSM, however, a 1000-fold higher concentrations of mOSM was required to stimulate proliferation of LIF dependent murine DA1a cells, differentiation of M1 macrophage cells, and inhibition of ES cell differentiation. On the other hand, mOSM inhibited growth of NIH3T3 cells at a 1000-fold lower concentration than that of hOSM. These results indicate that mOSM functions through a receptor which is distinct from that of the LIF receptor. Studies on the physiological role of OSM is underway. PMID- 9209422 TI - Receptor tyrosine kinases involved in hematopoietic progenitor cells. AB - To analyze the molecular mechanisms of the proliferation and differentiation of hematopoietic cells, we have cloned PTKs from sorted stem cells. We discuss the expression and function of receptor tyrosine kinases, STK/RON, TIE, TEK and HTK which have been cloned from these cells. STK and its ligand, MSP contributed to the motility and phagocytosis of peritoneal macrophages and bone absorption of osteoclasts. Apoptosis was induced in an erythroid cell line by the binding of MSP(MacrophageStimulating Protein). TIE, TEK and HTK were interestingly expressed in the subpopulations of stem cells and related to the myeloid differentiation. These study will indicate the heterogeneity of stem cells and their diverse differentiation. PMID- 9209423 TI - Analysis of factors controlling primary germ layer formation and early hematopoiesis using embryonic stem cell in vitro differentiation. AB - Differentiation and subsequent development are intricately interwoven processes operating as an integrated whole to form the organism. As an approach to examine these interactions in early mammalian development, we used embryonic stem (ES) cell in vitro differentiation. ES cells can, depending upon the environment differentiated to neuroectoderm, mesoderm and hematopoietic cells. We developed a serum-free, chemically defined medium (CDM) in which ES cells survive and differentiate. In CDM, in the absence of exogenous factors, ES cells form neuroectoderm, upregulating the early neural marker Pax-6. This is consistent with the view that neuroectoderm development can represent a default state, where the absence or sequestration of mesoderm inducing factors permits neuroectoderm formation. In contrast, if CDM is supplemented with bone morphogenetic protein (BMP) 2 or 4 a process resembling primitive streak formation, least at the molecular level occurs, with the formation of mesoderm and subsequently endothelial and hematopoietic cells. If used with care, ES cell in vitro differentiation can act as a guide in understanding the environment which controls early differentiation events in mammals. PMID- 9209424 TI - In vitro analysis of potency restriction during epiblast differentiation. AB - During mammalian gastrulation, posterior part of primitive ectoderm differentiates into subsets of mesoderm cells, the most proximal portion of which subsequently gives rise to vascular endothelial cells and hematopoietic cells. To analyze this process in which the potency of multipotent epiblast cells is restricted progressively into hematopoietic cell lineages, we have established a culture system that allows differentiation of epiblast cells as well as embryonic stem cells into hematopoietic cells. Using this culture system, we showed that all parts of epiblast tissues at early streak and late bud stages preserved hematogenic potency, while it was lost from anterior region at early head fold stages. However, this loss of hematogenic potency in the anterior region of the head fold stage embryo was reinduced by addition of activin in the culture. Moreover, in order to detect transitory stages of mesoderm cells that are the direct progeny of multipotent epiblast cells, we developed three monoclonal antibodies that are able to define distinct subsets of mesoderm cells in the gastrulating embryo. These results are discussed from a view of cell-mass based commitment. PMID- 9209425 TI - Self-renewal of hematopoietic progenitor cells under defined condition. PMID- 9209426 TI - Stage-specific cell-cycling of hematopoietic progenitor cells. AB - We studied the growth of hematopoietic progenitors at different progressive stages of differentiation and focused especially on changes in cell-cycling. Hematopoietic progenitors from 5-fluorouracil (5-FU)-treated mice were separated into three groups on the basis of differentiation, Stages I, II, and III, and have studied their cell-cycling. Primary marrow cells collected from 5-FU-treated mice were categorized as Stage I progenitors. Stages II and III progenitors are early and late progenies of Stage I progenitors, respectively. The rate of growth of hematopoietic progenitors supported by interleukin-3 (IL-3) and steel factor (SF) was estimated by sequential analysis of colony formation and studying replating efficiency of individual colonies. The time required for hematopoietic progenitors to go through the cell-cycle shortened as their stage of differentiation progressed. Similar results were obtained with other growth factor combinations. The analysis of DNA content of cells suggests that shortening of cell-cycling is mainly due to a decrease in the time of G1 phase of the cell-cycle. Our results demonstrate that in early hematopoiesis, the cell cycling of hematopoietic progenitors accelerates as they differentiate. PMID- 9209427 TI - Cascade regulation of cytokine production in granulopoiesis. AB - Injection of IL-3 producing T cells (STIL-3) resulted in a granulocytosis in the syngeneic mice. A high IL-3 activity was detected in the culture supernatant of the spleen cells of these mice, but only a low activity was detectable in the bone marrow cell-conditioned medium. There was no significant difference in the distribution of the STIL-3 cells between the spleen and the bone marrow of the mice injected with the STIL-3 cells. Two possibilities have been envisaged from these observations; i) IL-3 induces production of granulocyte stimulating cytokines (CSFs) from hemopoietic cells hence resulting in the granulocytosis in the STIL-3 injected mice, ii) an inhibitor of IL-3 is produced in response to an excess stimuli with IL-3 in the bone marrow. We found that IL-3 induced a production of IL-6, G- and GM-CSF from bone marrow cells. In contrast, stimulation of the bone marrow cells with an excess level of IL-3 resulted in a production of a heat-labile activity (NIL-3) antagonistic to IL-3. Furthermore, stimulation of bone marrow cells with IL-6, G- or GM-CSF did not induce the production of IL-3, indicating a hierarchical regulation of the cytokine production. These observations have provided positive evidences to the above mentioned 2 possibilities, and indicate the existence of a positive feedback mechanism in the IL-3-induced granulocytosis, as well as the presence of a negative feedback mechanism for the homeostatic regulation of the granulopoiesis. PMID- 9209429 TI - Molecular mechanisms involved in long-term maintenance of erythroleukaemia cells by stromal cells. PMID- 9209428 TI - In vitro long-term culture of human primitive hematopoietic cells supported by murine stromal cell line MS-5. AB - When Lin-CD34+CD38- cells from normal human cord blood were cocultured with MS-5, colony forming cells were maintained for over 8 weeks. Prevention of contact between MS-5 and Lin-CD34+CD38- cells by using a membrane filter was negligible for this activity, indicating that the activity of MS-5 on human primitive hematopoietic cells may be due to soluble factor(s) secreted from MS-5. We tried to purify this activity by a [3H]TdR incorporation assay. The activity was found in 150 kD fraction and was neutralized with anti-mSCF (stem cell factor) antibody. Another 20-30 kD fraction synergized with mSCF to stimulate the growth of Lin-CD34+CD38- cells but failed alone. This fraction supported the growth of the G-CSF (granulocyte-colony stimulating factor)-dependent cell line FD/GR3, FDC P2 transfected with mG-CSF receptor cDNA. This synergy was canceled in the presence of soluble mG-CSF receptor. Addition of anti-mSCF antibody and soluble mG-CSF receptor to the culture completely abrogated the activity of MS-5-culture supernatant. These results indicate the activity of MS-5 on Lin-CD34+CD38- cells is due to synergistic effect of mSCF and mG-CSF. PMID- 9209430 TI - Chloroquine induces basophilic differentiation of HL-60 cells. AB - Many agents have been known to induce the differentiation of HL-60 cells. However, only a small number of reports on the basophilic differentiation of this cell line are known. In this study we show that the exposure of HL-60 cells to chloroquine induces to differentiate into basophils. This chloroquine-induced change suggests that the increase in intracellular pH and the upregulation of p21 with subsequent downregulation of cdc2 kinase are triggers for basophilic differentiation of this cell line. PMID- 9209431 TI - Induction of apoptosis by overexpression of the PU. 1/Spi-1 gene in murine erythroleukemia cells. PMID- 9209432 TI - Establishment and characterization of 7,12-dimethylbenz[a]anthracene (DMBA) induced rat erythroleukemia cell lines. AB - To investigate molecular mechanisms and biological behaviors of 7,12 dimethylbenz[a]anthracene (DMBA)-induced rat erythroleukemia, we established 8 new culture cell lines from 7 primary erythroleukemias. We designated them KYD 10, 12, 17, 32, 38, 44A, 44B, and 49. Representative clones isolated from each cell line in early passage were analyzed cytogenetically and genetically. All cell lines except KYD-12 possessed the specific N-ras mutation at the 2nd base of codon 61. Four of them showed #2 trisomy (KYD-10, 32, 38, 44B), and the rest normal diploid karyotype (2n). KYD-32 cells showed Robertson type II #2 trisomy which had never been clonally isolated in vitro although it was reported in some DMBA leukemias in vivo. We further studied the genomic imbalance related to the N ras allele using mutant-allele-specific amplification (MASA) method. Deletion of normal N-ras allele was found in 5 of 8 cell lines. KYD-32 and 38 retained the normal N-ras allele. The specific N-ras mutation and allelic loss of wild type N ras were correlated with advanced cell proliferation in culture probably independent of #2 trisomy. PMID- 9209433 TI - A new type-II membrane protein in erythropoietic organs enhances erythropoiesis. AB - Erythropoiesis is regulated by erythropoietin and microenvironment of erythropoietic organs such as fetal liver and spleen in mice. We developed in vitro erythropoietic microenvironment by established stromal cell lines from erythropoietic organs, in which proliferation and differentiation of erythroid progenitor cells are supported by direct cell-to-cell contact between erythroid progenitor cells and the stroma cells. To identify the functional molecules of the stromal cells, we established monoclonal antibodies which detected specific molecules of the erythropoietic organs. Among those monoclonal antibodies, 11 D exhibited specific immunohistochemical staining of the red pulp of spleen where erythropoiesis dominates. Using this monoclonal antibody, we cloned a new gene (SMAP-1) which codes a type-II membrane protein. Over expression of its antisense cDNA in a stroma cell line caused a decrease in the erythropoietic supporting activity. Thus, SMAP-1 may play an important role in the erythropoietic stimulatory activity of the stromal cells. PMID- 9209435 TI - The p53-deficient hemopoietic stem cells: their resistance to radiation apoptosis, but lasted transiently. AB - Hemopoietic stem cells lacking with p53 are known to be resistant to radiation apoptosis: p53-product induces, on one hand, the cells that have been injured by radiation to stop cell-cycle at G, phase, and on the other hand the cells that have not been repaired to go into apoptosis. Thus, in the p53-deficient mice, the hemopoietic stem cells after graded dose of radiation show a flatter survival curve for radiation, i.e., like a curve for radio-resistant phenotype. However, we found the radio-resistance in the stem cell was only transient, and an apoptosis in p53-deficient hemopoietic stem cells, we made concentrated analysis including the end-labeling technique to detect double strand breaks of DNA. Results clearly showed that, according to the end-labeling for spleen colonies, the p53-deficiency showed rather higher incidence of apoptosis (30-50%) as compared with the wild-control (17%). Irradiation with 200cGy for the recipient mice two months after transplantation, showed an induction of higher incidence of hemopoietic malignancies, when the heterozygous marrow was repopulated, however, none of increase was observed in the recipient mice repopulated with the homozygous bone marrow. Clear difference between the hetero- and the homozygous in inducing hemopoietic malignancies with the 200cGy-radiation may be reflected by the different degree of delayed appearance of apoptosis during the development of the spleen colonies. PMID- 9209434 TI - MafB represses erythroid genes and differentiation through direct interaction with c-Ets-1. AB - Using a yeast interaction screen with a DNA-bound Ets-1 protein we have identified MafB as a direct interaction partner. This distant AP-1 relative, which is specifically expressed in myelomonocytic cells, binds to the DNA binding domain of Ets-1 via its basic region/leucine zipper domain. MafB represses Ets-1 transactivation of synthetic promoters containing Ets binding sites in yeast as well as avian cells. Both Ets-1 and Maf family proteins have been implicated in erythroid specific gene expression. Here we show that MafB inhibits Ets-1 mediated transactivation of the transferrin receptor, which is essential for heme synthesis and erythroid differentiation. Consequently, overexpression of MafB in an erythroblast cell line down-regulates the endogenous transferrin receptor gene and inhibits the cells' potential to differentiate into erythrocytes, without affecting cellular proliferation. PMID- 9209436 TI - Role of the stem cell factor (SCF) receptor and the alternative forms of its ligand (SCF) in the induction of long-term growth by stroma cells. AB - Self renewal and differentiation of hematopoietic stem cells is regulated by the hematopoietic microenvironment. The Stem Cell Factor (SCF) has been shown to be one of the essential factors that governs stem cell maintenance. In this abstract we describe a functional analysis of the membrane bound and soluble SCF within the context of stroma cocultures with hematopoietic cells. We report that transmembrane SCF is essential for long term growth, whereas soluble SCF supports only short term proliferation of stroma dependent hematopoietic cells. We also show that the SCF/c-kit interaction can be substituted by an unknown mechanism. To determine the molecular mechanism involved in maintenance of hematopoietic cells on stroma we analyzed the c-kit expression during coculture. We demonstrate that c-kit expression was downregulated during coculture. Downregulation was induced by the coculture itself and not by external factors. PMID- 9209437 TI - Development of erythroid cells from mouse embryonic stem cells in culture: potential use for erythroid transcription factor study. AB - We developed an efficient differentiation induction system from mouse embryonic stem (ES) cells into blood cells by coculture on a novel stromal cell line named OP9, in order to analyze molecular mechanisms of hematopoietic cell development and differentiation. ES cells could give rise to adult type definitive erythrocytes, myeloid and B lineage cells via multipotential hematopoietic precursor cells, when the cells were simply cocultured with the OP9 stromal cells. The temporal pattern of the appearance of erythroid lineage cells during the differentiation induction was very similar to that detected in mouse ontogeny. This differentiation induction method should facilitate to dissect the function of erythroid transcription factors during erythroid lineage cell development. PMID- 9209438 TI - Of the GATA-binding proteins, only GATA-4 selectively regulates the human IL-5 gene promoter in IL-5 producing cells which express multiple GATA-binding proteins. AB - Interleukin-5 (IL-5) is produced by T lymphocytes and known to support B cell growth and eosinophilic differentiation of the progenitor cells. Using ATL-16T cells which express IL-5 mRNA, we have identified a region, within the human IL-5 gene promoter, that regulates IL-5 gene transcription. This cis-acting sequence contains the core binding motif, (A/T)GATA(A/G), for GATA-binding family proteins and thus suggests the involvement of these family members. In this report, we describe the cloning of human GATA-4 (hGATA-4) and show that hGATA-4 selectively interacts with the -70 GATA site within the IL-5 proximal promoter region. By promoter deletion and mutation analyses, we established this region as a positive regulatory element. Cotransfection experiments revealed that both hGATA-4 and PMA/A23187 stimulation are necessary for the IL-5 promoter activation. The requirement of another regulatory element called CLE0, which lies downstream of the -70 GATA site, was also demonstrated. ATL-16T cells express mRNA of three GATA-binding proteins, hGATA-2, hGATA-3 and hGATA-4, and each of them has a potential to bind to the consensus (A/T)GATA(G/ A) motif. However, using ATL-16T nuclear extract, we demonstrated that GATA-4 is the only GATA-binding protein that forms specific DNA-protein complex with the -70 GATA site. The electrophoretic mobility shift assay with extracts of COS cells expressing GATA binding proteins showed that GATA-4 has the highest binding affinity to the -70 GATA site among the three GATA-binding proteins. When the transactivation ability was compared among the three, GATA-4 showed the highest activity. These results demonstrate the selective role of GATA-4 in the transcriptional regulation of the IL-5 gene in a circumstance where multiple members of the GATA-binding proteins are expressed. PMID- 9209439 TI - Molecular mechanism of blastic crisis in chronic myelocytic leukemia. AB - The t(3;21)(q26;q22), which is usually found in blastic crisis of chronic myelocytic leukemia or myelodysplastic syndrome-derived leukemia, produces an AML1/EVI-1 fusion protein of 180 kD containing amino-terminal half of AML1 including a runt homology domain which is fused to the entire of zinc finger EVI 1 protein. Thus, AML1/EVI-1 fusion protein is a chimeric transcription factor including a runt homology domain from AML1 and two zinc finger domains from EVI 1, totally three DNA binding domains, and an acidic domain from EVI-1. The AML1/EVI-1 fusion protein possesses the dual functions, namely, differentiation block and stimulation of proliferation. The ability of differentiation block depends on the runt homology domain in the AML1 part and the effect to stimulate proliferation depends on the second zinc finger domain in the EVI-1 portion. The AML1/EVI-1 could play an important role in leukemic progression of chronic myelocytic leukemia by these dual functions as a transcription factor. PMID- 9209440 TI - Randomized studies with interferon in chronic myelogenous leukemia (CML) and comparative molecular aspects. German CML Study Group. AB - Four randomized prospective studies on interferon alpha (IFN) in CML report varying degrees of prolongation of the chronic phase of CML and of survival as compared to conventional therapies. There is agreement that IFN prolongs survival as compared to standard busulfan. There is disagreement, however, as to which degree IFN is superior to hydroxyurea. Whereas the randomized studies of the Italian cooperative group and of the British MRC find a statistically significant survival advantage of IFN over hydroxyurea of about 20 months, this difference is only 10 months in the German randomized study and not significant. One reason for this difference might be the more intensive treatment schedule for the hydroxyurea control group in the German study. Other reasons might be differences in risk profiles between the patient groups studied and in strategies of IFN therapy. About 1% of the human genome consists of retroviral or retroviral-like sequences. By analogy to animal models, endogenous retroviruses might also have pathogenic potential in human disease. The transposon-like structure of retroviruses that enables them to integrate at almost any position in the host genome and the capability of retroviruses to serve as efficient vehicles of cellular genes are in support of a pathogenic potential. Furthermore, particles resembling retroviruses have been observed long ago in human embryonic and malignant tissues and cell lines. Sequence information and the transcriptional activity of the endogenous sequences argue against the possibility that these sequences are only fossil relics of early evolutionary periods. Most of the sequences appear to be inactivated by stop codons or frameshifts, making the genomic localization of open reading frames with biological activity difficult. Up to now, mutagenesis by insertion of retroviral-like sequences in sporadic cases of human disease appears to be the only example of pathogenic relevance of retroviruses in man. PMID- 9209441 TI - Donor chimaerism is a strong indicator of disease free survival following bone marrow transplantation for chronic myeloid leukaemia. AB - Although Chronic Myeloid Leukaemia (CML) can be treated successfully with allogeneic bone marrow transplantation (BMT), leukaemia relapse remains a significant clinical problem. Molecular monitoring of the post transplant marrow can be useful in predicting relapse particularly in CML patients where the Philadelphia chromosome or its molecular counterpart, the BCR-ABL fusion messenger RNA can be used as a leukaemia specific marker of minimal residual disease (MRD). We have investigated chimaerism (using polymerase chain reaction of short tandem repeat sequences (STR-PCR)) and MRD status (using reverse transcriptase PCR of the BCR-ABL fusion mRNA) in a serial fashion in 18 patients who were in clinical and haematological remission post allogeneic BMT for chronic phase CML. Eleven patients exhibited complete donor chimaerism with no evidence of minimal residual disease. Five patients had transient or low level stable MC. Late MC and MRD was observed in two patients who relapsed > 6 years after T cell depleted BMT for CML. Thus STR-PCR is an appropriate screening test in the post transplant setting for CML patients, but those patients exhibiting mixed haemopoietic chimaerism should also be monitored using a leukaemia specific sensitive molecular assay. PMID- 9209442 TI - B cell malignancy and hepatitis C virus infection. AB - Italian authors report that hepatitis C virus (HCV) infection may be one of the causes of lymphoid malignancy such as non-Hodgkin's lymphoma (NHL) and Waldenstrom's macroglobulinemia (WM). To assess the relationship between HCV infection and B cell malignancy (BCM) in Japan, we analyzed HCV-RNA in 50 patients with BCM [25 cases of NHL, 4 of WM and 21 of multiple myeloma (MM)] and determined genotype of infected HCV(Okamoto's classification) using reverse transcription-polymerase chain reaction assay. Eight (16.0%) of 50 patients with BCM were HCV-RNA positive [HCV(+)], while no patients were HCV(+) in control group (18 patients of non-B cell NHL). Numbers of HCV(+) cases in each group examined were as follows; four (16.0%) in B cell NHL (genotype II/III/IV were 3/1/0, respectively), one (25.0%) in WM (genotype III) and three (14.3%) in MM (genotype II/III/IV were 1/1/1, respectively). All patients examined had no symptoms and signs suggesting vasculitis. The incidence of HCV infection in the patients with BCM was markedly higher than that (approximately 1%) of healthy blood donors in Japan. We also experienced four B cell NHL cases with splenic or hepatic origin in the course of chronic hepatitis C. These results implicate the association between persistent HCV infection and the occurrence of BCM. PMID- 9209443 TI - Molecular events in chronic myeloid leukemia progression. AB - Chronic myelogenous leukemia presents two distinct clinical phases: the chronic phase is characterised by a marked expansion of the myeloid compartment which still retains a normal differentiative capacity, whereas a differentiation block is the clinical hallmark of the acute transformation. The molecular mechanism underlying the CML progression are still poorly understood. The occurrence of additional molecular lesions, involving the p53, the RAS and the p16 genes may complement and fulfil the BCR/ABL transforming potential, finally leading to an acute leukemic phenotype. However, several lines of evidence suggest that also quantitative changes of the BCR/ABL transcript amounts could explain the progression of the leukemic phenotype in the BCR/ABL-positive hematologic malignancies. PMID- 9209444 TI - Cardiac involvement with lymphoma/leukemia: a report of three autopsy cases. AB - The clinicopathological findings in two patients with ATL and one patient with promyelocytic Leukemia affecting the heart are described. All three cases tested positive for HTLV-I while one of the ATL cases was also positive for HIV. Cardiac infiltrate in the ATL cases varied from microscopic focii in one case to macroscopic infiltration mimicking myocardial infarction in the other case. There were microabscesses in the myocardium in the case afflicted with promyelocytic Leukemia. All three cases had extensive malignant disease with multiple organ infiltration. There was no ante-mortem manifestation of cardiac involvement in any case. The cardiac dimensions were all within the normal range. The possible pathophysiology of cardiac involvement with Lymphoma/Leukemia is discussed. PMID- 9209445 TI - Role of an immunosuppressive cytokine, interleukin-10, in the development of pyothorax-associated lymphoma. AB - Malignant lymphoma frequently develops in the pleural cavity of the patients with long-standing pyothorax. Thus, the term pyothorax-associated lymphoma (PAL) has been proposed for this type of tumor. Most of PALs are diffuse lymphoma of B cell type and contain Epstein-Barr virus (EBV) DNA. We have established two lymphoma cell lines from the biopsy specimens of PAL cases, OPL-1 and OPL-2. Both cell lines contain EBV DNA, but only OPL-1 expresses Epstein-Barr virus nuclear antigen 2 (EBNA2) that works as a target molecule for cell-mediated immune response. In this study, we examined the expression of immunosuppressive factors in OPLs. Only OPL-1, not OPL-2, expressed interleukin-10 (IL-10) mRNA and secreted IL-10 into culture supernatant. Both OPL-1 and OPL-2 expressed transforming growth factor (TGF) beta 1 mRNA, however, neither expressed latent TGF beta binding protein (LTBP) mRNA at detectable level by Northern blot analysis. Because TGF beta expresses its functions in cooperation with LTBP, the biological functions of TGF beta 1 could be negligible. Neither cell lines expressed EBV BCRF1 mRNA at detectable level, a viral gene product which is partly homologous to human IL-10 and shares biological activities of IL-10. Since OPL-1 shows weaker proliferative activity than OPL-2 and expresses viral antigens, the production of an immunosuppressive cytokine, IL-10, might contribute to the development of overt lymphoma. The present study suggested that immunosuppressive cytokine plays a role in lymphomagenesis of immunocompetent patients. PMID- 9209446 TI - Matrix metalloproteinases and their inhibitors in acute myeloid leukemia. PMID- 9209447 TI - Effective prevention of thrombocytopenia using adenovirus-mediated transfer of HST-11FGF-4 gene: in vivo and in vitro studies. AB - A novel biological function of HST-1 protein (FGF-4) was investigated by constructing an adenovirus vector containing the HST-1 cDNA and applied for thrombocytopenia as a gene therapy. A single intraperitoneal injection of the replication-deficient adenovirus containing the HST-1 gene (Adex1HST-1) into mice caused a two-fold increase in peripheral platelet count for 30 days, and effectively prevented experimentally induced thrombocytopenia. Studies of Adex1HST-1-infected or HST-1 protein-treated megakaryocytic Dami cells suggested that HST-1 protein promotes megakaryocyte maturation, and increases cytokine secretion from megakaryocyte and adhesive interactions between megakaryocyte and endothelial cells. Colony assay revealed that HST-1 protein stimulated CFU-MK (colony-forming unit of megakaryocyte) not alone but synergistically with early acting cytokines such as IL-3 or Tpo (c-mpl ligand) as a megakaryocyte potentiating factor. These results have important implications for clinical application of the Adex1HST-1 for thrombocytopenia. PMID- 9209449 TI - Immunostaining of PRAD1/cyclin D1 protein as a marker for the diagnosis of mantle cell lymphoma. AB - The overexpression of PRAD1/cyclin D1 gene activated by the 11q13 translocation and its molecular counterpart BCL-1 rearrangement is frequently associated with mantle cell lymphomas (MCLs). Recently, we produced a monoclonal antibody, 5D4, against the PRAD1/cyclin D1 product, and demonstrated that the positive nuclear staining by this antibody correlates with PRAD1/cyclin D1 mRNA overexpression in MCLs. In the present study, we have immunohistochemically examined the cyclin D1 protein in a large series of 315 malignant lymphomas including 39 MCLs on paraffin sections. The nuclear positive pattern was found in 35 (90%) of 39 MCLs with an exceptional case of immunocytoma among the B-cell lymphomas examined. In the other cases, the positivity was absent or appeared to lie within the cytoplasm without nuclear staining. We therefore propose that the immunolocalization of cyclin D1 protein is an essential marker for the definite diagnosis of MCL. PMID- 9209448 TI - Genetic aberrations in the development and subsequent progression of myelodysplastic syndrome. AB - We performed longitudinal analyses of chromosomes and studied the configuration of NRAS, TP53, NF1, and cFMS genes in 70 patients with myelodysplastic syndrome(MDS). The NRAS mutations were detected in 6 patients(9%) at codons 12 or 13. The TP53 mutations were found in 10 patients(14%) in exons 4 through 8. Longitudinal studies revealed that the NRAS mutation was a late-appearing event, while the TP53 mutations were detectable at the presentation of MDS. No patients had both NRAS and TP53 mutations, simultaneously. NF1 and cFMS genes showed any mutational event among these 70 patients. Patients with a TP53 mutation had a significantly shorter survival time than those with an NRAS mutation or those without NRAS or TP53 mutation. However, patients who showed an NRAS mutation had a shorter survival time once the mutation emerged, similar to that of patients with a TP53 mutation. PMID- 9209450 TI - Anaplastic large cell lymphomas expressing the novel chimeric protein p80NPM/ALK: a distinct clinicopathologic entity. AB - Some anaplastic large cell lymphomas (ALCLs) carry a specific chromosomal translocation, t(2;5)(p23;q35). Recently, we found a novel hyperphosphorylated 80 kDa protein tyrosine kinase, p80, in ALCLs with t(2;5). Subsequent cDNA cloning revealed p80 to be a fusion protein of two genes, the novel tyrosine kinase gene and the nucleophosmin gene, in accordance with the sequence of the NPM/ALK gene (Morris et al.). Meanwhile, the clinicopathologic features of p80-carrying ALCLs have remained unclear. Paraffin sections of 105 cases of ALCL were immunostained using anti-p80 antibody, and 30 of them were shown to express p80. Clinicopathological comparison between p80-positive and -negative ALCLs revealed that p80-positive cases occurred in a far younger patient age group and the patients showed a far better 5-year survival rate. These data showed that p80 positive ALCL is a distinct entity both clinically and pathogenetically, and should be differentiated from p80-negative ALCL. PMID- 9209451 TI - Monitoring the efficiency of interferon-alpha therapy in chronic myelogenous leukemia (CML) patients by competitive polymerase chain reaction. AB - Interferon alpha (IFN-alpha) induces cytogenetic responses of variable degree in patients with CML. We sought to establish the relationship between BCR-ABL transcript numbers measured by competitive two-step reverse transcription polymerase chain reaction (RT-PCR) and cytogenetic status in CML patients treated with IFN-alpha. All 398 samples from 163 patients investigated by RT-PCR were positive for BCR-ABL transcripts. In order to standardize results for variability in RNA and cDNA quality, we quantified total ABL transcripts in each sample as internal control. The BCR-ABL/ABL ratios correlated with the cytogenetic results. Quantitative nested PCR allowed the detection of residual BCR-ABL transcripts in all complete cytogenetic responders on IFN-alpha. We conclude that competitive PCR with internal controls is a reliable method for monitoring patients on IFN alpha and reduces the need for repeated marrow investigations. PMID- 9209452 TI - Dual phasic suppression of viral replication following de novo human immunodeficiency virus type 1 (HIV-1) infection in lymphocytes of asymptomatic HIV-1 carriers. AB - Replication of human immunodeficiency virus type 1 (HIV-1) is suppressed in asymptomatic HIV-1 carriers (ACs). By using an in vitro experimental system, the mechanism of this suppression was investigated. Following in vitro infection of a laboratory HIV-1 strain, the peripheral blood mononuclear cells (PBMC) of ACs transiently supported a low level of viral replication, then the virus production rapidly decreased. PCR analysis revealed that HIV-1 proviral DNA integrated in the PBMC of ACs following infection gradually decreased. Such tapering consequences of in vitro HIV-1 infection in the PBMC of ACs were abrogated by depletion of CD8+ T cells from the culture. Furthermore, the viruses subsequently produced by the PBMC of an AC were less able to replicate than the virus produced by CD8+ cell-depleted PBMC of the same donor. These observations suggested that the CD8+ T cell-mediated suppression of HIV-1 replication in ACs may involve both cytocidal and cytostatic mechanisms: the former kills the cells producing viruses, and the latter inhibits viral spread by reducing viral infectivity. PMID- 9209454 TI - In vivo drug-selectable markers in gene therapy. AB - Two-gene vectors with positive or positive-negative drug-selectable markers enable the expansion or elimination of gentetically modified cells in vivo. We have established a bicistronic retroviral vector system which utilizes an internal ribosome entry site (IRES) to co-express two independent genes with high efficiency. As a positive-negative (suicide) marker, Herpes simplex virus thymidine kinase was co-expressed with the human multidrug resistance gene, MDR1. Using this vector, almost all the MDR1-transduced cells showed hypersensitivity to a nucleoside analog, ganciclovir. As a dominant selectable marker, the MDR1 gene was co-expressed with alpha-galactosidase A for the model of gene therapy of Fabry disease. Vincristine selection efficiently enhanced the population of transduced cells expressing the second non-selectable genes. These drug selectable retroviral vectors could be applicable to the therapy of many diseases. PMID- 9209453 TI - Intracellular pharmacodynamics of ara-C and flowcytometric analysis of cell cycle progression in leukemia chemotherapy. AB - To establish the most effective and reasonable mode of combining and administrering ara-C with other antileukemic agents in chemotherapy for acute leukemia, the action mechanisms of ara-C was investigated in terms of intracellular pharmacodynamics and the biochemical action mechanism of ara-C was investigated in leukemic cell. Rensonable methods of administering the agent was considered as follows. 1. A low level of ara-C in the incubation medium induced a higher concentration of ara-CTP in leukemic cells. Therefore, maintenance of even a low plasma ara-C level after ara-C therapy could enhance the antileukemic effect of the agent. 2. Ara-C activation was increased in the presence of 6MP by suppressing elevation of deaminase activity in the cell suspection medium. Therefore, administration of 6MP prior to ara-C therapy could enhance the antileukemic effect of the agent. 3. Ten micrograms/ml of ara-C, corresponding to intermediate dose ara-C therapy, induced rapid endonuclease activation, DNA ladder fragmentation and subsequent apoptosis in large numbers of leukemic cells, suggesting that intermediate dose ara-C therapy is effective in reducing residual leukemic cells after therapy. 4. Blood transfusion for patients with high grade anemia prior to bebenoyl ara-C therapy prolonged higher and longer plasma drug maintenance. 5. Flowcytometry of cell cycle progression of L1210 cells treated by ara-C and daunorubicin revealed that a combination of ara-C first and daunorubicin second was superior to the reverse sequential combination. These improvements in the mode of administering ara-C could provide better results following chemotherapy for leukemia. PMID- 9209455 TI - Functional relationships among ETS gene family members. PMID- 9209456 TI - Manipulation of T cell response to tumors by targeting on costimulatory pathway. AB - T cells require at least two signals to be fully activated: one is generated by interactions between antigen-specific receptor on T cell and peptide-MHC complexes on tumor cells and second signal is delivered by costimulatory molecules on antigen presenting cells to their counter-receptor on T cells. We demonstrated previously that expression of T cell costimulatory molecule B7-1, a counterreceptor for CD28, on tumors led to tumor regression in syngeneic mice. We have used retrovirus to transfer B7-1 into a variety of murine tumor lines to examine their ability to stimulate CTL in vivo and in vitro. Expression of B7 results in increased immunogenicity in immunogenic, but not poorly-immunogenic tumors, suggesting a deficiency of tumor cells on antigen presentation. We analyze tumor epitopes associated with MHC molecules by HPLC combining with specific CTL clones and the results indicate that many non-immunodominant epitopes do not normally induce a response unless B7 costimulation is provided. Furthermore, increased T cell receptor signaling, such as co-expression of CD2 ligand with B7-1, can convert some poorly-immunogenic tumours to become immunogenic. Our results indicate that deficiency on antigenic signaling in many tumors could be a quantitative phenomenon. Induction of T cell immunity by targeting on both antigen receptor and costimulatory pathway thus may be useful for cancer treatment. PMID- 9209457 TI - The anti-tumor effects of IL-12 involve enhanced IFN-gamma production by anti tumor T cells, their accumulation to tumor sites and in situ IFN-gamma production. AB - We investigated cellular and molecular mechanisms underlying the anti-tumor effects of IL-12. Intraperitoneal injections of rIL-12 into later stages of tumor bearing mice induced not only a striking reversal of suppressed IFN-gamma production by splenic T cells, but also complete regression of s.c. growing tumors. A massive infiltration of lymphoid cells was found following IL-12 treatment. Whereas fresh spleen cells obtained from IL-12-treated tumor-bearing mice failed to express IFN-gamma mRNA, significant levels of IFN-gamma mRNA were detected in the tumor mass of the same mice. The systemic administration of anti IFN-gamma antibody (Ab) prior to IL-12 injection abrogated the anti-tumor effect of IL-12 although this Ab did neither inhibit accumulation of lymphoid cells to tumor sites nor in situ IFN-gamma expression. Importantly, while high levels of inducible nitric oxide synthase (iNOS) mRNA expression was induced in tumor masses after IL-12 treatment, its expression was completely inhibited by pretreatment with anti-IFN-gamma Ab. Thus, induction of tumor regression by IL-12 is ascribed to a series of events: a striking reversal of suppressed IFN-gamma production by anti-tumor T cells, their accumulation and IFN-gamma production at tumor sites, and manifestation of IFN-gamma activity as exemplified by iNOS expression. PMID- 9209458 TI - Immunoregulation by B7 and IL-12 gene transfer. AB - Our recent studies using various costimulatory molecules have demonstrated that antitumor effect could be induced by B7- or B70-transduced mouse tumors. To augment antitumor effect in vivo, the combination therapy with a costimulatory gene and a cytokine, interkeukin 12 (IL-12), gene to treat metastatic mouse lung tumor was investigated. We transfected with mouse B7 and/or IL-12 into mouse lung carcinoma 3LL, and three transfectants (IL-12/3LL, B7/3LL and IL-12/B7/3LL) were generated. CTL activity induced by the inoculation of IL-12/B7/3LL was increased about 10-fold compared with parental 3LL inoculation. We then examined the therapeutic efficacy of combination with B7 and IL-12-transduced tumors. Four weeks after 3LL inoculation, lung metastasis was significantly reduced by IL 12/B7/3LL post-inoculation, indicating that potent therapeutic antitumor immunity can be induced by combination with costimulators B7 and IL-12. Recently, it was reported that p40 subunit of IL-12 appeared to be a specific inhibitor for IL-12 heterodimer in vitro. To clarify the biological functions of p40 in vivo, we generated the myoblast transfectants which produced IL-12 p40 alone. Local production of IL-12 p40 from transfectant could suppress allogenic CTL induction and Th1-type antibodies (IgG2a/2b/3) production in vivo. Furthermore, IL-12 p40 producing myoblast are less susceptible to rejection compared with parental myoblast, indicating that IL-12 p40 gene transfer may be useful therapeutically in Th1-mediated transplantation and autoimmune disorders. PMID- 9209459 TI - Protective and therapeutic immunity against leukemia induced by irradiated B7-1 (CD80)-transduced leukemic cells. AB - B7 molecules provide an important costimulatory signal for T cell receptor/CD3 mediated T cell activation via binding to their cognate receptors, CD28 and CTLA 4. We have introduced B7-1 (CD80) into M1 cells, spontaneously-occurred mouse myelocytic leukemic cells and assessed its potential in the induction immunity to leukemia cells. Syngeneic, immunocompetent SL mice receiving polyclonal B7-1 transduced M1 cells showed prolonged survival than control mice. Two independent B7-1-transduced monoclonal sublines, M1-B7-1+ (F20) and M1-B7-1+ (F7), were rejected in 100% an 50% of SL mice, respectively. In vivo depletion of T cell subsets showed that both CD4+ and CD8+ T cells were indispensable for the B7-1 dependent anti-leukemic immunity. Although a single exposure to irradiated monoclonal M1-B7-1+ cells were not fully effective, multiple exposures induced protective immunity against subsequent challenge with M1 cells. Furthermore, hyperimmunization with irradiated monoclonal M1-B7-1+ (F7) cells could partly cure mice previously injected with a lethal number of M1 cells. Although other groups have demonstrated that live, proliferating B7-1-transduced leukemic cells can improve antitumor immunity, this is the first report which shows that irradiated B7-1-transduced myeloid leukemic cells can induce protective and therapeutic immunity against leukemia. PMID- 9209460 TI - HTLV-1 uveitis (HU). AB - We reported HTLV-1 uveitis (HU) as the third distinct clinical entity associated with HTLV-1 infection. Uveitis has advantages for studying the pathogenetic mechanisms of inflammatory diseases caused by HTLV-1, since observation of the disease process is relatively easy and clinical samples from the lesion can be obtained. Results of our clinical, virological and epidemiological studies of HU patients will be first summarized. Then we present results on in vitro studies that showed transactivation of HTLV-1 LTR by thyroid hormone and discuss the significance in the context of pathogenesis of HTLV-1-related inflammatory diseases. PMID- 9209461 TI - Cytogenetics of primary testicular nonseminoma, residual mature teratoma, and growing teratoma lesion in individual patients. AB - Residual mature teratoma (RMT) is often left behind when metastases of primary nonseminomatous germ cell tumors (NSs) are treated with chemotherapy. RMT is composed of fully differentiated somatic tissue. A growing teratoma (GTE) lesion may occur after (incomplete) resection of RMT. To shed light on tumor progression or the mechanism(s) of therapy related differentiation we investigated the chromosomal pattern of the primary NSs and RMTs in twelve patients, of the primary NS, RMT, and GTE lesion in one patient, and of the RMT and GTE lesion in two patients. Although several chromosomal differences are observed between the RMT and NSs and between the GTE and RMTs in the same patient, we obtained no evidence that specific chromosomal alteration(s) play a role in metastasis or differentiation. PMID- 9209462 TI - A novel chromosomal abnormality involving chromosomes 2 and 18 in a patient with myelodysplastic syndrome. AB - Cytogenetic analysis of bone marrow cells from a patient with myelodysplastic syndrome associated with eosinophilia showed a complex translocation with a 46,XY,t(2;18;2)(p23;q11;q32) karyotype. The patient has refractory anemia (RA) according to the French-American-British Cooperative Group (FAB) classification, and after 90 months of follow-up he shows no evidence of leukemic transformation. This chromosomal abnormality has not been previously described in myelodysplastic syndromes and may be associated with good prognosis as the patient has been stable for a long time. PMID- 9209463 TI - Isochromosomes of both short and long arms of chromosome 12 resulting in an additional copy of chromosome 12 in a case of splenic lymphoma with villous lymphocytes. AB - We report a case of splenic lymphoma with villous lymphocytes showing a karyotype with an isochromosome for both the long arm and the short arm of chromosome 12, i(12)(p10) and i(12)(q10), effectively resulting in trisomy 12. This is, apparently, the first documented case of an additional copy of chromosome 12 resulting from isochromosome formation in a neoplastic disorder. PMID- 9209464 TI - Cytologic characterization of two distinct alpha satellite DNA domains on human chromosome 7, using double-labeling hybridizations in fluorescence and electron microscopy on a melanoma cell line. AB - The most prominent class of centromeric DNA sequences belongs to the alpha satellite family of tandemly repeated DNA. The human chromosome 7 has been shown to contain two distinct alpha satellite arrays: D7Z1 and D7Z2, separated by 1 Mb. The order of these arrays was analyzed in normal blood cells and in the melanoma cell line IPC182 with two approaches using in situ hybridization: (1) Relative mapping on high-resolution chromosomes in fluorescence and electron microscopy (EM); and (2) simultaneous visualization of the two sequences using fluorochromes of different colors or gold particles of different sizes. The location within the centromeric area of chromosome 7, on the side of the short arm for D7Z2 and near the long arm for D7Z1 is confirmed. In addition, the hybridization signal of D7Z2 is confined to two small areas of the centromeric region in external positions, whereas the D7Z1 signal covers the entire width of the primary constriction. In situ hybridization with D7Z1 and D7Z2, performed on the melanoma cell line IPC 182, allowed characterization of two isochromosomes, i(7)(q10) and idic(7)(q11), as well as the der(7)t(7;12) observed in this cell line. The three-derived chromosomes appeared to result from different breakpoints, but only D7Z1 was conserved in all cases, suggesting the importance of this sequence for the centromeric function. PMID- 9209465 TI - Characterization by FISH of a t(5;13) in a patient with bilateral retinoblastoma. AB - We have used fluorescence in situ hybridization (FISH) with a biotinylated cosmid probe (13q14) to screen 25 unrelated cases with bilateral retinoblastoma and one case with unilateral retinoblastoma. In 25 cases no deletion or chromosome rearrangements were, found. One constitutional mutation resulting from a de novo balanced chromosome translocation (5;13) in a patient with bilateral retinoblastoma was detected. PMID- 9209466 TI - Minimal residual disease in acute monocytic leukemia patient with trisomy 11 and partial tandem duplication of MLL. AB - We studied MLL rearrangements in five patients with myeloid hematologic malignancies with trisomy 11. Two had acute monocytic leukemia (AMoL), one had chronic myelomonocytic leukemia, one had refractory anemia, and the other had juvenile chronic myelogenous leukemia. Only one patient, a 15-year-old boy with AMoL and simple trisomy 11, showed rearrangement of MLL. He did not respond to chemotherapy, and successfully underwent bone marrow transplantation, but suffered a relapse 22 months later. Reverse transcription-polymerase chain reaction (RT-PCR) and sequencing analyses of bone marrow cells revealed a tandem duplication of MLL, and his relapse was predictable by sequential RT-PCR studies before it was clinically evident. Of 16 acute myeloid leukemia patients with trisomy 11 and rearrangement of MLL reported, our patient was the youngest in age and the only one with AMoL. PMID- 9209468 TI - Pigmented choroid plexus carcinoma: a cytogenetic and ultrastructural study. AB - A pigmented choroid plexus carcinoma was studied. The pigment was Fontana positive, and the neoplastic cells focally expressed melanosomal marker HMB45 and contained probable aberrant melanosomes. The tumor was composed of two pseudodiploid clones, having the karyotypes 46,XY,inv(4)(q12q35),t(6;15)(q21;q22),inv(7)(p11.2q22),t(19; 22) (q13.4;q11.2)[15]/46,XY,t(4;14)(q31.1; p11.2),t(12;13)(p11.1;q34)[6]. The available data seem to indicate that rearrangements of 7p11-12, 9q11-12, 15q22, and 19q13.4 may play a role in the development of choroid plexus carcinomas. PMID- 9209467 TI - Cytogenetic and molecular genetic characterization of a chromosome 2 rearrangement in a case of human papillary thyroid carcinoma with radiation history. AB - Karyotype analysis of a primary culture from a case of papillary thyroid cancer (PTC) showed an abnormal short arm of one homologue of chromosome 2 as sole abnormality in 4 of 16 metaphases. Based on G-banding analysis, two different aberration types on chromosome 2 could be assumed representing either a del(2)(p22-23) or a pericentric inversion. Further comparative genomic hybridization (CGH) analysis as well as fluorescence in situ hybridization (FISH) analysis were performed to confirm the assumed alterations. While CGH analysis showed no loss of chromosome 2 material, FISH with yeast artificial chromosome (YAC) probes homologous to the region 2p22-23 demonstrated two pericentric inversions of chromosome 2 involving different breakpoints on 2p in 6.8% and 4.2% of the metaphases, respectively. Polymerase chain reaction (PCR) analysis with degenerated oligonucleotide primers that bind within the conserved catalytic domain of tyrosine kinase (tk) genes resulted in amplification products with DNA of YAC 851D11 suggesting the presence of such genes at or near the translocation breakpoint. PMID- 9209469 TI - Clonal chromosome aberrations in three of five sporadic angiomyolipomas of the kidney. AB - Clonal chromosomal changes were detected in three of five sporadic angiomyolipomas of the kidney irrespective of a solitary or multifocal appearance of this benign tumor type. No specific chromosomal changes have been identified. Including the cytogenetic data of the four renal angiomyolipomas reported in the literature, trisomy 7 as the single clonal chromosomal abnormality was detected in two angiomyolipomas. Because trisomy 7 has been reported in both neoplastic and nonneoplastic kidney cells, it may be assumed that trisomy 7 is already physiologically resident in renal cells but undergoes positive selection in this tumor type. PMID- 9209470 TI - Constitutional inversion of chromosome 7 and hematologic cancers. AB - Nonrandom aberrations of chromosome 7 have been described in various hematopoietic disorders. We describe here two unrelated families with the same constitutional inversion of chromosome 7 [inv(7)(q11.2q22)]. The probands in both families had acute leukemia and cytogenetic analysis revealed that the inversion was the sole cytogenetic abnormality in the bone marrow at diagnosis. There is a history of hematologic diseases in one of these families that included a son who is a carrier of this constitutional inversion. The distal inversion breakpoint lies within the common region of chromosome loss identified in some myeloid diseases. These observations raise the possibility that this inherited chromosome rearrangement could result in a mutation of a tumor suppressor gene and possibly represent a predisposing event for the development of leukemia in these individuals. PMID- 9209471 TI - Establishment and characterization of a renal cell carcinoma line from a patient with von Hippel-Lindau syndrome. AB - It is not known how the von Hippel-Lindau (VHL) gene and the, as yet unidentified, renal cell carcinoma (RCC) gene(s) interact to result in RCC; nor is it known if mutations in both, or all genes are necessary for this progression. The availability of a RCC cell line from a VHL patient would be useful in studies comparing sporadic RCC with RCC resulting from VHL syndrome; and for determining the relationship or interaction of the RCC gene with the VHL gene to produce a common tumor type. This paper describes the isolation and characterization of a renal cell carcinoma cell line derived from a patient with von Hippel-Lindau disease. The line is epithelial in origin and the genome contains a familial mutation in the VHL gene. Tissue culture studies indicate that this cell line, although immortalized, is not fully transformed. Chromosomal analysis performed on cells derived from disseminated primary tumor cells revealed no detectable chromosomal abnormalities. However, analysis performed on cells at passages 9, 19, 41, and 79 showed both numerical and structural chromosomal changes. The cytogenetic profile of this cell line demonstrated a number of abnormalities known to be associated with RCC from patients with and without VHL syndrome. PMID- 9209472 TI - Chromosomal abnormality inv(3)(q21q26) associated with multilineage hematopoietic progenitor cells in hematopoietic malignancies. AB - We have identified ten patients with acute myeloid leukemia (AML) and one patient with chronic myeloid leukemia with megakaryocytic crisis who displayed an inv(3)(q21q26). Seven of them had an additional monosomy 7. Most of them had a myelodysplastic syndrome (MDS) preceding AML, normal or increased platelet counts, increased number of megakaryocyte, megakaryocytic dysplasia, and erythroid dysplasia. There was a high incidence of resistance to induction chemotherapy, short remission time, and early relapse. Seven patients were immunologically analyzed. The main immunophenotypes were as follow: CD7+, CD34+, HLA-DR+, CD38+, CD13+, CD33+, CDw65+, CD2-, CD3-, CD4-, CD8-, CD19+, CD20-, CD11b . Our results suggest that the leukemia with inv(3)(q21q26) represents a new cytogenetic-clinicopathologic subtype, characterized by 1) abnormal megakaryopoiesis and multiple hematopoietic lineage involvement; 2) an antecedent MDS; 3) poor response to conventional chemotherapy; and 4) expression of CD7, CD34, CD38, HLA-DR, CD13, and CD33 antigens. We propose that the malignant transformation in patients with inv(3)(q21q26) occurs in an early stem cell prior to lineage commitment. PMID- 9209473 TI - Regional cancer cytogenetics: a report on 1,143 diagnostic cases. AB - The results of studies from a regional cancer cytogenetics diagnostic service are reported. In a 10-year period, 1,143 marrow samples from patients with newly diagnosed leukemia and myelodysplastic syndrome were referred. Successful studies were completed on 992 cases (87%). Among all referred cases, the rates of detection of cytogenetically abnormal clones were 95% for chronic myelogenous leukemia (CML), 54% for acute lymphoblastic leukemia (ALL), 51% for acute myeloid leukemia (ANLL), and 43% for myelodysplastic syndrome (MDS). Of 169 cases of CML studied, 90.5% bore the standard Philadelphia chromosome (Ph), 3.55% had an unusual Ph, and 5.33% were Ph-negative. Among the 59 cases of cytogenetically abnormal MDS, common abnormalities observed were trisomy 8 and changes resulting in loss of material from the long arm of chromosomes 5 and 7, and 20q-. Of the 168 abnormal ANLL, there was a strikingly non-random pattern of aneuploidy, with monosomy 7 and trisomy 8 predominating. Common structural changes observed were changes resulting in loss of material from the long arm of chromosomes 5 and 7, trisomy 8, rearrangements of 11q23, t(15;17), t(8;21), rearrangements of 12q13 and 3q, inversion 16, trisomy 11, Ph, trisomy 21, t(6;9) and t(1;22). The differences between adult and pediatric findings were minor, with the exception of chromosome 5 abnormalities, which were common among adults with ANLL but rare in the pediatric cases. There were 273 ALLs with abnormal cytogenetic findings. There was preferential gain of chromosomes 21, X, 14, 6, 4, 18, 17, and 10 (in decreasing order of frequency) in leukemic clones. Of the 193 ALLs with structural changes, many fell into-well-defined categories with established correlations to FAB subtypes. Common changes in ALL were rearrangements of 9p, 12p, 6q, TCR loci, 11q23, Ig loci, and 8q24, and duplication of 1q, Ph, i(17q), t(1;19), i(9q) and dic(9;12). The detailed documentation of the cytogenetic findings in this relatively large, single-institution study will likely facilitate the further characterization of rare, primary cytogenetic changes associated with leukemias and MDS. From a managed health care perspective, regional cancer cytogenetic services may be cost-effective alternatives to single institution laboratories. PMID- 9209474 TI - Involvement of the long arm of chromosome 9 in medulloblastoma in an adult. AB - Medulloblastoma is the most common primitive neuroectodermal tumor (PNET) in children, but is very rare in adults. An isochromosome for the long arms of 17, i(17q), is found in about 30% of pediatric cases. Cytogenetic studies in adults are very scarce; only six cases have been described cytogenetically: three cases had normal karyotype, two were studied partially, and another presented only two clonal structural anomalies: del(9)(q12) and del(11)(q22). We studied the chromosomes from medulloblastoma in a 27-year-old woman and found one hypotetraploid stemline with clonal alterations. In the structural anomalies, chromosomes 3, 9, 12, and i(17q) were involved. Chromosome 9 presented a deletion in the long arm, del(9)(q13), with consequent loss of the 9q13-->qter region. This anomaly was similar to one found in a previous case. We suggest that the partial loss of the long arm of chromosome 9 may be a characteristic change of adult medulloblastoma. PMID- 9209475 TI - Deletion of the long arm of chromosome 7 in lipoma. PMID- 9209476 TI - Variant Philadelphia translocation t(9;17)(q34.2-3;q21.3) with colocalization of the BCR and ABL genes on chromosome 9 in chronic myeloid leukemia. PMID- 9209477 TI - Penicilloyl peptides are recognized as T cell antigenic determinants in penicillin allergy. AB - Although hapten immune responses have been intensively studied in the mouse, very little is known about hapten determinants involved in human allergic reactions. Penicillins, as chemically reactive compounds of low molecular weight, constitute typical examples of hapten allergens for humans. Penicillins become immunogenic only after covalent binding to carrier proteins and in this form frequently induced IgE-mediated allergic reactions in patients subjected to antibiotic treatment. However, our previous data strongly indicated that penicillins also form part of the epitopes contacting the antigen receptors of beta lactam specific T cells in allergic individuals. We have therefore investigated the molecular constraints involved in the T cell immune response to penicillin G (Pen G). Designer peptides containing a DRB1*0401-binding motif and covalently modified with Pen G via a lysine epsilon-amino group were found to induce proliferation of Pen G-specific T cell clones. A precise positioning of the hapten molecule on the peptide backbone was required for optimal T cell recognition. Furthermore, we extended these observations from our designer peptides to show that a peptide sequence derived from a natural DRB1*1101-binding peptide modified in vitro with Pen G, also acquired antigenic properties. Our data for the first time provide insight into the manner in which allergenic haptens are recognized by human T cells involved in allergic reactions to drugs and suggest possible mechanisms leading to the onset of these adverse immune responses. PMID- 9209478 TI - Diverse transcription factors are involved in the quantitative regulation of transcriptional activation of kappa promoters. AB - Immunoglobulin kappa promoters show sequence divergence but conserved function between different subgroups. Here we show that three separate 5' elements are required for synergistic stimulation of transcription with the decamer in a kappa promoter. These sites are a 5' E-box, a 3' AT-rich region in the pentadecamer (pd) element, and the kappa-Y element. Elf-1 is a novel kappa-Y element ligand induced upon mitogenic stimulation of resting B lymphocytes. Furthermore, the 5' E2A-like E-box in the pd element could be substituted by an upstream stimulatory factor motif with conservation of function. Thus, the synergistic activation requirements of kappa transcription is strictly dependent on the quantitative presence of transcription factor-binding motifs 5' of the decamer, but these differ qualitatively in that they may bind an array of proteins with conserved function. PMID- 9209479 TI - Induction of CD4+ T cell depletion in mice doubly transgenic for HIV gp120 and human CD4. AB - It has been suggested that loss of uninfected T cells in HIV infection occurs because of lymphocyte activation resulting in cell death by apoptosis. To address the question of whether cross-linking of CD4/HIV gp120 complexes by antibodies were sufficient to induce T cell depletion in vivo, we developed an animal model of continuous interaction between human CD4 (hCD4), gp120 and anti-gp120 antibodies in the absence of other viral factors. Double-transgenic mice have been generated in which T cells express on their membrane hCD4 and secrete HIV gp120. Although these mice have hCD4/gp120 complexes present on the surface of T cells, they do not show gross immunological abnormalities, and they are able to produce anti-gp120 antibodies following immunization with denaturated gp120. However, double-transgenic mice with antibodies to gp120, when immunized with tetanus toxoid, mount an IgG response that is significantly lower than that of double-transgenic mice without antibodies to gp120. Furthermore, the presence of anti-gp120 antibodies leads to CD4+ T cell depletion and immunodeficiency in the absence of HIV infection. Thus, the antibody response to gp120 can lead to CD4+ T cell attrition in vivo. PMID- 9209480 TI - Maturation of Peyer's patch dendritic cells in vitro upon stimulation via cytokines or CD40 triggering. AB - In mouse Peyer's patches (PP), dendritic cells (DC) are localized in T cell areas as NLDC145+ CD11c+ cells, and in the dome and corona region of the follicle as NLDC145- CD11c+ cells, respectively, suggesting the presence of two different DC populations with distinct roles in antigen uptake, processing, and presentation. However, it is not clear how this relates to DC maturation. In this report, we demonstrate that freshly-isolated CD11c+ DC have the properties of immature DC since they endocytose soluble antigens, phagocytose particulate material such as latex beads, synthetize major histocompatibility complex (MHC) class II and invariant chain, but, at the same time, display low stimulatory activity for resting T cells, as shown in mixed-lymphocyte reaction and oxidative mitogenesis assays. When cultured for 24 h in the presence of the cytokines granulocyte macrophage colony-stimulating factor and tumor necrosis factor or anti-CD40, the cells undergo dramatic phenotypic and functional changes characteristic of DC maturation. After 24 h stimulation in vitro, CD11c+ cells lose the ability to take up proteins such as ovalbumin, and in parallel with this decline, the biosynthesis of MHC class II and invariant chain is dramatically down-regulated or eliminated. On the other hand cells treated in vitro exhibit on the cell surface higher levels of MHC class II, of co-stimulatory molecules (CD80, CD86), of adhesion molecules (CD44, intercellular adhesion molecule-1), and acquire expression of the interdigitating DC surface marker NLDC145. Concomitantly, the ability to stimulate naive T cells drastically increased after in vitro treatment with both stimuli. Taken together, our results indicate that the majority of DC in the PP are immature in terms of their antigen-uptake capacity. These sentinel antigen presenting cells are strategically positioned at the dome region of PP, where antigens are transcytosed via the M cells from the gut lumen. A second population of mature interdigitating NLDC145+ CD11c+ DC stimulates naive unprimed T cells in interfollicular areas by up-regulation of surface ligands and accessory signals. PMID- 9209481 TI - HLA-B27 modulates intracellular survival of Salmonella enteritidis in human monocytic cells. AB - Human major histocompatibility complex class I allele HLA-B27 is associated with a group of diseases called spondyloarthropathies. In reactive arthritis (ReA), the disease is triggered by certain infections, e.g. gastroenteritis caused by Salmonella. The host/microbe interaction is abnormal in susceptible individuals leading to inefficient elimination of arthritis-triggering bacteria, fragments of them, or both, after the initial infection. Using transfected human monocytic U937 cell lines, we demonstrate that the expression of the HLA-B27 antigen does not influence the uptake of S. enteritidis into U937 cells in vitro. Interestingly, HLA-B27 remarkably impairs the elimination of S. enteritidis within the HLA-B27 transfected U937 cells. The impaired elimination of ReA triggering microbes by HLA-B27+ monocytes may offer an explanation for the persistence of ReA-triggering microbes in susceptible HLA-B27+ individuals. This modulation of the host/microbe interaction by HLA-B27 may have an important role in the pathogenesis of ReA. PMID- 9209482 TI - P-selectin glycoprotein ligand-1 mediates rolling of mouse bone marrow-derived mast cells on P-selectin but not efficiently on E-selectin. AB - It has been shown recently that mast cells play an essential role as a source of tumor necrosis factor-alpha production during neutrophil recruitment to sites of bacterial infection. Increased numbers of mast cells are indeed noted at sites of wound healing and inflammation. These cells are either recruited from the bone marrow or proliferate locally under cytokine stimulation. Little is known about how mast cell progenitors extravasate into tissue. Using antibody-like fusion proteins of mouse E-selectin and P-selectin, we have analyzed the ability of immature mouse bone marrow-derived mast cells (BMMC) to interact with the endothelial selectins. The P-selectin glycoprotein ligand-1 (PSGL-1) was affinity isolated from detergent extracts of surface biotinylated BMMC with both selectin IgG fusion proteins. However, only P-selectin-IgG, but not E-selectin-IgG showed significant interaction with intact BMMC as tested by flow cytometry and cell attachment assays with the immobilized fusion proteins under flow and non-flow conditions at physiological shear stress. Thus, in spite of carrying the necessary carbohydrate modifications which enable solubilized PSGL-1 to bind avidly to E-selectin, PSGL-1 on the surface of BMMC is presented in a way that prevents it from interacting efficiently with E-selectin. Affinity-purified rabbit antibodies against mouse PSGL-1 almost completely blocked the interaction of BMMC with P-selectin-IgG in flow cytometry as well as in cell adhesion assays under static and under flow conditions. Our data reveal that PSGL-1 is the major binding site for P-selectin on mouse BMMC progenitors, but does not support efficient interactions with E-selectin. PMID- 9209483 TI - The T/B cell interaction involved in induction of the mouse IgG2ab suppression is restricted by major histocompatibility complex class I, but not class II molecules. AB - To determine the major histocompatibility complex (MHC) restriction of the T/ B cell interaction involved in a negative regulation of Ig production, we used mouse model of T cell-induced IgG2ab suppression in vivo. Normal or specifically triggered T splenocytes from mice of the Igha haplotype, when neonatally transferred into histocompatible Igha/b heterozygotes, are able to induce a specific and total suppression of the IgG2ab allotype. Nevertheless, only transfer of IgG2ab-primed Igha T splenocytes induces this suppression in Ighb/b homozygous congenic mice in which the whole IgG2a isotype production is inhibited. This suppression is chronically maintained by CD8+ T cells, but can be experimentally reversed. We have established that the suppression induction required a CD4+CD8+ T cell cooperation and operated via the recognition by the involved TCR of C gamma 2ab-derived peptides presented by the target B cells in an MHC haplotype-restricted manner. Here, by using Ighb mice genetically deficient for MHC class I (beta 2-microglobulin%, or beta 2m%) or class II (I-A beta%) molecules, we demonstrate functionally that the suppression induction implicates an MHC class I-, but not class II-restricted interaction. Indeed, the anti-IgG2ab T cells transferred into Ighb H-2b I-A beta% mice carry out the suppression process normally, while in Ighb H-2b beta 2m% recipients, their suppression induction capacity is significantly inhibited. Moreover, the C gamma 2ab 103-118 peptide, identified as the sole C gamma 2ab-derived peptide able to amplify the anti-IgG2ab T cell reactivity in Igha H-2b mice, is also able to stabilize the H-2Db, but not the H-2Kb class I molecules at the surface of RMA-S (TAP2-, H-2b) cells. These results indicate that, despite the CD4+/CD8+ T cell cooperation during the induction phase of suppression only MHC class I molecule expression is required at the surface of IgG2ab+ B cells for suppression establishment. PMID- 9209485 TI - L-selectin ligands in rat high endothelium: multivalent sialyl Lewis x glycans are high-affinity inhibitors of lymphocyte adhesion. AB - Lymphocyte homing is initiated by their tethering to and rolling on the high endothelium and is followed by extravasation into the lymph nodes. We show here that glycosylated cell adhesion molecule-1 (GlyCAM-1), CD34, and sialyl Lewis x (sLex) are present on rat lymph node high endothelium analyzed by using monoclonal antibodies. alpha (1,3)fucosyltransferase VII (Fuc-TVII), the last enzyme involved in the synthesis of the sLex sequence is also expressed on the rat lymph node high endothelium. We have synthesized a family of sLex-decorated oligosaccharide structures and used them to inhibit lymphocyte binding to high endothelium in the Stamper-Woodruff assay. Monovalent sLex, branched di- and tetravalent sLex, as well as a linear tetravalent sLex significantly reduce lymphocyte binding to endothelium. The branched and linear forms of tetravalent sLex were clearly superior inhibitors of the L-selectin-dependent lymphocyte adhesion, with IC50 values in low nanomolar range. In contrast, the fucose-free analogs having the same charge and approximately the same size as the corresponding sLex glycans had no effect on lymphocyte binding and served as negative controls. Taken together, these data show the crucial importance of sLex in the endothelial ligands for L-selectin. Furthermore, we suggest that L selectin acts as an oligomer on the lymphocyte surface as it binds multivalent sLex glycans. PMID- 9209484 TI - TAP-dependent major histocompatibility complex class I presentation of soluble proteins using listeriolysin. AB - Immunization of mice with mixtures of listeriolysin, a pore-forming hemolysin secreted by the pathogenic bacterium Listeria monocytogenes, together with soluble ovalbumin, nucleoprotein of influenza virus, or beta-galactosidase of Escherichia coli, resulted in strong cytotoxic CD8 T cell responses to each of the respective passenger proteins in vivo. Also, the concomitant addition of either protein with listeriolysin to target cells elicited efficient sensitization of these cells which could be attributed to the pore-forming activity of listeriolysin. This response was dependent upon a functional TAP transporter and was inhibitable by brefeldin A, indicating the transfer of the soluble proteins into the cytosol and the classical major histocompatibility (MHC) class I presentation pathway. The treatment of target cells with listeriolysin under our experimental conditions did not affect cell viability and the pores generated by listeriolysin treatment were repaired within 60 min. Introduction of soluble proteins into the MHC class I presentation pathway by listeriolysin provides a powerful system to study the cytotoxic response towards intracellular pathogens and would allow for rapid screening of potential antigens in vaccine formulations. PMID- 9209486 TI - Analysis of the requirement for beta 2-microglobulin for expression and formation of human CD1 antigens. AB - Human CD1 form a group of nonpolymorphic leukocyte surface molecules with homology to major histocompatibility complex (MHC) proteins. Recent findings in human and in mouse demonstrate the capacity of CD1 molecules to present nonpeptide components like lipids or lipoglycans as well as peptides. We studied the involvement of beta 2-microglobulin (beta 2m) in expression of the classic human CD1 proteins CD1a, CD1b, and CD1c. The beta 2m-deficient human melanoma cell line FO-1 was transiently transfected with either CD1a, CD1b, or CD1c DNA alone, or in combination with beta 2m using the adenovirus-enhanced receptor mediated transfer infection system. Only co-transfection of FO-1 cells with CD1+ beta 2m resulted in the detection of CD1 Ag by monoclonal antibodies (mAb). This indicated that CD1 mAb recognized determinants are dependent on beta 2m and raised the question whether beta 2m-free forms of CD1 can be expressed. Therefore, to visualize CD1 molecule expression independently of beta 2m, we expressed tagged recombinant forms. A full-length CD1b construct tagged at the very C terminus with a small peptide was transported to the plasma membrane only when beta 2m was co-transfected. beta 2m involvement in the transport of CD1 was confirmed by expression of soluble forms of CD1a, CD1b, and CD1c in three different cell types. Analogous to tagged full-length CD1b, secretion of the soluble CD1 constructs was strictly dependent on beta 2m. The soluble CD1 chimeras were secreted as complexes with endogenous beta 2m. Thus, similar to its role for MHC class I expression, beta 2m is essential for processing and surface transport of the classic human CD1 molecules CD1a, CD1b, and CD1c. PMID- 9209487 TI - Interleukin-15-induced maturation of human natural killer cells from early thymic precursors: selective expression of CD94/NKG2-A as the only HLA class I-specific inhibitory receptor. AB - Immature postnatal thymocytes were shown to contain precursors which, under suitable culture conditions, give rise to phenotypically and functionally mature natural killer (NK) cells. Here, we analyzed the effect of different cytokines for their ability to induce the expression of HLA class I-specific inhibitory receptor(s) during the process of NK cell development from immature thymocytes. From thymocyte cell suspensions depleted of CD2+, CD3+, CD4+, CD8+, CD56+, and CD16+ cells, we further removed cells expressing HLA class I-specific inhibitory receptors including CD94/NKG2-A, p58.1, and p58.2 by immunomagnetic bead separation. The resulting cells did not contain any of the above NK receptors as determined by immunofluorescence and flow cytometric analysis, as well as by reverse transcriptase polymerase chain reaction (RT-PCR) amplification using appropriate sets of primers. Although different cytokines have been used, including interleukin (IL)-7, stem cell factor (SCF), IL-2, and IL-15, only IL-2 or IL-15 induced cell proliferation when used alone. Moreover, maturation towards CD3- CD56+ cells displaying cytolytic activity against the HLA class I- targets K562 or 221 was detectable in cultures containing IL-15 used alone or in combination with IL-7 or SCF. On the other hand, these CD3- CD56+ cell populations did not lyse HLA class I+ target cells, including autologous PHA blasts. Analysis of the expression of the various HLA class I-specific inhibitory NK receptors revealed the presence of high proportions of CD94/ NKG2-A+ cells, while the NK receptors belonging to the Ig superfamily were undetectable both by immunofluorescence and by RT-PCR analysis. The expression of CD94/NKG2-A appeared to be responsible for the inability of cells to lyse HLA class I+ target cells. Thus, addition of anti-CD94 monoclonal antibodies of IgM isotype resulted in lysis of autologous target cells. The use of 221 cells transfected with different HLA class I alleles as target cells confirmed the broad class I specificity of CD94/NKG2-A receptor. Our experiments indicate that IL-15 provides an appropriate stimulus to the expression of CD94/NKG2-A, but not of other class I-specific NK receptors in the process of maturation of NK cells from thymocyte precursors. PMID- 9209488 TI - Age-related impaired affinity maturation and differential D-JH gene usage in human VH6-expressing B lymphocytes from healthy individuals. AB - To elucidate the basic molecular events underlying humoral immunity during ontogeny and senescence, we analyzed a panel of 179 polymerase chain reaction derived VH6-D-JH rearrangements from cord blood, peripheral blood, and spleen. Nucleotide sequence analysis of the CDR3 region shows that there is a difference in D and JH gene usage in functional rearrangements between lymphocytes from peripheral blood and spleen. Analysis of the VH6 gene shows that the mutational frequencies rise from 0.81% in cord blood to 1.96% in peripheral blood lymphocytes derived from young adults, and decrease to 0.80% in samples from individuals older than 50 years. The number of rearrangements carrying mutations follows a similar pattern: 22% in cord blood, 73% in the age group 20-49 years, and 57% in the age group over 50 years. The mutational frequencies among the mutated genes are, however, similar for cord blood and young adults, 2.76% and 2.51%, respectively, and 1.3% in older adults. These data show an age-related impaired affinity maturation which might relate to the decrease in immunological responsiveness among the elderly. PMID- 9209489 TI - A retro-inverso analog mimics the cognate peptide epitope of a CD4+ T cell clone. AB - Synthetic analogs of peptide epitopes may activate specific T helper cells, antagonize their antigen receptors, or block recognition by competing for major histocompatibility complex (MHC) class II binding sites. Rationally designed peptides may therefore prove useful as vaccines and for treatment of autoimmune diseases and allergies mediated by CD4+ T cells. However, their susceptibility to proteolytic degradation limits the applicability of conventional peptides in vivo. By contrast, retro-inverso analogs, in which a native sequence is substituted with D-amino acids linked with a reversed backbone, resist proteolysis and still maintain the side chain topology of the corresponding natural peptide. We report here that an end group-modified retro-inverso analog of the IgG2ab heavy chain allopeptide determinant gamma 2ab 435-447 was recognized by an I-Ad-restricted, gamma 2ab 435-447-reactive T cell clone. The pseudopeptide elicited near-maximal interleukin-2 responses, although 300-fold higher concentrations were needed than the native determinant. The weaker antigenicity of the retro-inverso analog could be fully accounted for by an impaired I-Ad binding capacity, which might reflect reduced ability of the distorted main chain to form hydrogen bonds with I-Ad. Glycine substitution at the residue corresponding to the first primary anchor (P1) of the native peptide abrogated I-Ad binding and antigenicity of the retro-inverso analog. Thus, the pseudopeptide resembled the native determinant with respect to orientation in the class II binding site, configuration of the epitopic side chains, and the constraints that governed the interactions between a major anchoring side chain and I-Ad. In conclusion, proteolytically resistant compounds with predefined capacity to interact with MHC class II allelic products and T cell antigen receptors may be designed by retro-inverso modification of native determinants. PMID- 9209490 TI - Thymocytes and RelB-dependent medullary epithelial cells provide growth-promoting and organization signals, respectively, to thymic medullary stromal cells. AB - The thymic medulla is composed of distinct epithelial cell subsets, defined in this report by the reactivity of two novel antibodies, 95 and 29, raised against mouse thymic epithelial cell lines. These antibodies were used to probe the development of medulla in wild-type or mutant thymuses. In CD3 epsilon-deficient mice where thymocyte maturation is arrested at the CD4- CD8- stage, few scattered 95+ and 29+ epithelial cells are found. When few mature thymocytes develop as in CD3- zeta/eta mice, expansion and organization of 95+ but not 29+ cells, becomes detectable. In RelB-deficient mice, T cell maturation proceeds normally but negative selection is inefficient due to the lack of thymic medulla and dendritic cells. Strikingly, 29+ epithelial cells are absent and 95+ medullary epithelial cells are scattered throughout the thymus, intermingling with CDR1+ cortical epithelium. In chimeric mice lacking only dendritic cells, the corticomedullary junction persists and both 95+ and 29+ epithelial cells are localized in the medulla. These results suggest that two types of signals are required for development of thymic medulla. A growth signal depends upon the presence of maturing thymocytes, but organization of the thymic medulla requires the presence of activated 29+ medullary epithelial cells. PMID- 9209491 TI - Diffuse large cell lymphomas are derived from mature B cells carrying V region genes with a high load of somatic mutation and evidence of selection for antibody expression. AB - In the Revised European American Lymphoma (REAL) classification, several subtypes of high-grade lymphomas were combined in the entity diffuse large cell lymphoma (DLL). In the present study, a total of 19 cases of DLL (10 cases of centroblastic lymphoma, 5 cases of mediastinal B cell lymphoma, 2 cases of immunoblastic lymphoma, 1 case of T cell-rich B lymphoma and one case of large cell anaplastic lymphoma) were analyzed for somatically mutated immunoglobulin V region genes. Somatic mutations are acquired in the course of the germinal center (GC) reaction and are thus found in GC B cells and their descendants, i.e. memory B cells. The V gene sequences revealed that the tumor cells of all five subtypes of DLL harbored mutated V region genes and are thus derived from antigen experienced (post) GC B cells. This indicates that from the point of view of the stage of development of the tumor precursor, the combination of those five subtypes to one entity, i.e. DLL, seems reasonable. In some cases, an unusually high frequency of somatic mutations was detected. This may indicate that DLL are derived from GC B cells, which, due to transforming events, stayed in the GC for prolonged periods of time, thereby accumulating a high load of somatic mutation. An analysis of the mutation pattern suggests that the tumor clone or its precursor were selected for antibody expression while acquiring somatic mutations. The latter observation discriminates DLL from classical Hodgkin's disease, where we recently also observed a high load of somatic mutation within rearranged V region genes, but a frequent occurrence of crippling mutations. PMID- 9209492 TI - Differential expression of binding sites for chemokine RANTES on human T lymphocytes. AB - The C-C chemokine RANTES, a T lymphocyte chemoattractant, is considered an important mediator of inflammation, allergy, and host defense against HIV-1 infection. In this study, we investigated the modulation of binding of RANTES to T lymphocytes. Human peripheral blood CD3+ T cells, when freshly isolated from buffy-coat blood, expressed a considerable number of high-affinity binding sites for RANTES. These cells also showed significant chemotactic migration in response to RANTES in vitro. After 6-15 h incubation at 37 degrees C, the binding of RANTES, but not of macrophage inflammatory protein-1 alpha (MIP-1 alpha) or of monocyte chemotactic protein-3 (MCP-3), consistently increased. Scatchard analyses indicated that the number of binding sites for RANTES increased about threefold by 15 h without any change in the affinity. The increase in RANTES binding was no longer detected by 24 h. This increase in the specific binding was mainly attributable to CD4+ T cells and was not associated with increased chemotactic activity of these cells in response to RANTES. Incubation with anti CD3 antibody for 15 h markedly reduced the binding capability of T cells for RANTES and was associated with decreased chemotactic activity. On the other hand, when T cells were incubated with interleukin-2 (IL-2) for 1 week, the specific binding for all three C-C chemokines, RANTES, MIP-1 alpha, and MCP-3 was markedly increased in comparison to cells cultured in the absence of IL-2. These results suggest that the expression of binding sites on T cells for RANTES is differentially modulated, indicating the existence of novel receptors for RANTES that do not bind MIP-1 alpha. PMID- 9209493 TI - Differential response of CD4+ V7+ and CD4+ V7- T cells to T cell receptor dependent signals: CD4+ V7+ T cells are co-stimulation independent and anti-V7 antibody blocks the induction of anergy by bacterial superantigen. AB - V7 is a novel cell surface glycoprotein that is expressed on 25% of circulating T lymphocytes. This molecule appears to play a critical role in T cell activation based on the observation that a monoclonal anti-V7 antibody inhibits T cell receptor (TCR)-dependent interleukin-2 (IL-2) production and proliferation of T cells. In the current study, CD4+ V7+ and CD4+ V7- T cells were separated from one another and their response to various stimuli analyzed. Although there were only minor differences between the two subsets in the expression of activation/differentiation markers, including CD45RA and R0 isotypes, when exposed to immobilized anti-CD3 or anti-TCR antibodies in the absence of APC, CD4+ V7+ T cells alone produced IL-2 and proliferated vigorously. By contrast, CD4+ V7- cells responded poorly to such stimuli, but they recovered their capacity to respond if antigen-presenting cells (APC) or anti-CD28-antibody were added to the cultures. The enhancement of the V7- T cell response by APC appears to be related to augmentation of TCR signals because the effect could be blocked by antibodies against molecules on APC [major histocompatibility (MHC) class II, CD86] that are known to up-regulate such signals through their interaction with counter-receptors on T cells. To assess the role of V7 in a system independent of co-stimulation, CD4+ T cells were stimulated with the bacterial superantigens, staphylococcal enterotoxins A and B. The cells responded by proliferating and then becoming anergic. Addition of anti-V7 antibody at the initiation of culture with superantigen did not inhibit cellular proliferation but prevented T cells from becoming anergic, while addition of anti-CD28 antibody had no effect on either proliferation or anergy induction. These results indicate that V7 and CD28 mediate distinct intracellular signals and suggest that V7 functions to preserve T cell reactivity whether the stimulus is mitogenic or anergizing. PMID- 9209494 TI - Association between receptor density, cellular activation, and transformation of adhesive behavior of flowing lymphocytes binding to VCAM-1. AB - We investigated the ability of purified vascular cell adhesion molecule-1 (VCAM 1), adsorbed on plastic, to capture and immobilize flowing lymphocytes, and the dependence of adhesive behavior on activation of the counter-receptor, alpha 4 beta 1 integrin. This integrin/immunoglobulin interaction bound lymphocytes at a wall shear stress at which the beta 2-integrin family has previously been found ineffective (> 0.1 Pa), and whereas lymphocytes rolled on lower concentrations of VCAM-1 (10 micrograms/ml), they were stationary at high concentrations (100 micrograms/ml). Activation of alpha 4 beta 1 integrin by Mn2+ or by antibody 12G10 or treatment of lymphocytes with phorbol ester caused transformation to stationary adhesion, and increased binding significantly only at the lower concentrations of VCAM-1. We thus hypothesized that formation of a high density of ligand between VCAM-1 and alpha 4 beta 1 integrin actively transformed lymphocyte behavior. This concept was supported by the finding that the proportion of lymphocytes rolling on the higher concentrations of VCAM-1 increased if cells were pretreated with azide to block energy-dependent responses, or if intracellular Ca2+ was chelated. However, not all activation responses were equivalent: only phorbol ester induced marked spreading of immobilized cells, and if pretreatment was prolonged, this agent even reduced the efficiency of initial attachment of flowing lymphocytes. Azide treatment had no effect on transformation to stationary adhesion caused by Mn2+ or activating antibody. Thus, different forms of lymphocyte activation were identifiable: external modification of integrin converted rolling to stationary attachment, did not require ATP, and was reversible; high-density ligand binding induced an energy-dependent signal for conversion from rolling to stationary attachment, but not spreading; and protein kinase C activation promoted stationary attachment and spreading, but not necessarily capture. VCAM-1 is thus a versatile adhesion receptor capable of supporting all stages of leukocyte attachment, i.e. rolling, stationary, and spreading, and of ligand-induced transformation of adhesion, although an additional signal appears necessary to promote lymphocyte spreading and migration. PMID- 9209496 TI - T cell receptor alpha-chain tail is required for protein kinase C-mediated down regulation, but not for signaling. AB - Antigen stimulation through the T cell receptor (TCR) induces phosphorylation of the associated CD3 gamma delta epsilon- and zeta-chain cytoplasmic tails. These events lead to the induction of the intracellular signaling pathways with concomitant receptor down-regulation. The TCR is down-regulated from the cell surface by the activation of protein kinase, C (PKC) and subsequent serine phosphorylation of the CD3 gamma-chain. We report here that the TCR alpha-chain cytoplasmic tail is also necessary for PKC-mediated internalization of the TCR complex. The requirement for the TCR alpha-chain cytoplasmic tail is specific for internalization of the TCR complex, since down-regulation of CD4 is still intact in hybridoma cells expressing a tailless TCR alpha-chain. The absence of TCR internalization directly correlates with defective PKC-mediated phosphorylation of the CD3 gamma-chain. Despite deficient PKC-mediated TCR down-regulation, the tailless alpha beta TCR still transduces antigenic signals resulting in the production of interleukin-2. Although the TCR tails are not obviously required for signal transduction, the TCR alpha-tail may serve as a targeting domain for PKC-mediated down-regulation of the TCR complex. PMID- 9209495 TI - HTLV-I-infected T cells activate autologous CD4+ T cells susceptible to HTLV-I infection in a costimulatory molecule-dependent fashion. AB - A vigorous production of human T lymphotropic virus type I (HTLV-I)-infected CD4+ T cells is closely associated with the development of adult T cell leukemia (ATL) and neurological disease. However, the immunological mechanisms leading to generation of the HTLV-I-infected cells are not fully clarified. The modulation of CD80 and CD86 expression on the HTLV-I-infected cells and its physiological role in the interaction of infected CD4+ T cells with uninfected CD4+ T cells was examined. The HTLV-I-infected CD4+ T cell lines established from ATL patients and normal donors by infecting their CD4+ T cells with the virus expressed CD80, CD86, and HLA-DR, and induced a proliferation of autologous and allogenic CD4+ T cells. While the CD4+ T cells stimulated with the autologous HTLV-I-infected cells for 7 days expressed CD80 and CD86 but not HTLV-I gene products, they expressed HTLV-I gag antigen after 4 weeks. The interaction of HTLV-I-infected and -uninfected CD4+ T cells was profoundly suppressed by a combination of CD80 and CD86 monoclonal antibodies. These results suggest that the induction of CD80 and CD86 on HTLV-I-infected CD4+ T cells participates actively in the generation of the virus-infected progenitor cells. PMID- 9209497 TI - Activation of protein kinase C attenuates early signals in Fas-mediated apoptosis. AB - Activation of protein kinase C (PKC) has been reported to inhibit Fas (APO-1, CD95)-mediated apoptosis in different cellular systems. Human Jurkat leukemic T cells express the Fas antigen in the cell membrane and undergo apoptosis upon cross-linking by anti-Fas monoclonal antibodies (mAb). Cleavage of the apoptosis associated protease CPP32 and its substrate poly(ADP-ribose)polymerase are observed after the engagement of Fas antigen with mAb. In this report, we show that all these effects are substantially inhibited by the activation of PKC with a phorbol ester. Bisindolylmaleimide, an inhibitor of PKC, prevents phorbol ester induced down-regulation of Fas signaling. Inhibition of Fas-mediated cell death by phorbol ester is also observed in other human leukemic T cell lines. Cross linking of Fas antigen by mAb results in the rapid increase in tyrosine phosphorylation of several protein substrates which is further elevated in the presence of the protein tyrosine phosphatase inhibitor, orthovanadate. Furthermore, orthovanadate markedly enhances the cell death response to Fas mAb in different human leukemic T cell lines and human T cell blasts. These effects of orthovanadate on early tyrosine phosphorylation and cell death are clearly diminished by PKC activation. These results strongly suggest that tyrosine phosphorylation is involved in Fas signaling in apoptosis and that PKC plays a negative role in Fas-mediated apoptosis by counteracting at a very early stage the signals generated following cross-linking of this receptor. PMID- 9209498 TI - Conversion in vivo from an early dominant Th0/Th1 response to a Th2 phenotype during the development of collagen-induced arthritis. AB - Over the past decade, the central role of T cells in the process of collagen induced arthritis (CIA) has been extensively documented. The inflammatory features of CIA and its successful modulation after treatment in vivo with Th2 lymphokines, known to down-regulate proinflammatory cytokines, classify CIA as a Th1-mediated disease. However, no direct evidence for the presence of the different T helper subsets has been obtained. To identify the collagen-specific CD4+ T cell subset(s) developing during the course of CIA, lymph nodes from susceptible DBA/1 mice (H-2q) were harvested at different times after injection of bovine type II collagen in Freund's complete adjuvant and checked by enzyme linked immunospot assay for the production of interferon (IFN)-gamma and interleukin (IL)-4. The results clearly showed that type II collagen-specific T cells secreting either IFN-gamma, IL-4, or both, develop early in vivo, before the onset of arthritis: the number of IFN-gamma-secreting cells was already maximal 15 days after immunization, whereas more IL-4-secreting cells were found at day 30, just before the onset of clinical arthritis. Another strategy was to establish collagen-specific CD4+ T cell lines and sublines in vitro and to analyze their lymphokine secretion pattern. Lines generated 8 days after immunization displayed a mixed lymphokine secretion pattern characteristic of Th0 cells or of a mixture of Th1 and Th2 cells. After limiting dilution of a day 8 line, 60% of the growing sublines were Th0-like (secreting IFN-gamma, IL-4, and IL-5), and 25% were Th1 (secreting IFN-gamma). By day 25 post-immunization, 33% of the generated sublines were Th0-like, 11% Th1, and 56% Th2 (secreting IL-4 and IL-5). Moreover, all the sublines raised from the lymph nodes of arthritic mice harvested at day 55 secreted high amounts of Th2 lymphokines, and only 3 out of 14 also produced some IFN-gamma. This study demonstrates that during the course of CIA the collagen-specific CD4+ T cell response shifts in vivo from a dominant Th0/Th1 response to a clear Th2 phenotype. These results contribute to our understanding of the collagen-specific CD4+ T helper subsets which develop during the induction and clinical phases of CIA. PMID- 9209499 TI - Identification of epitopes on a mutant form of Pseudomonas exotoxin using serum from humans treated with Pseudomonas exotoxin containing immunotoxins. AB - PE38 is a 38-kDa derivative of the 66-kDa Pseudomonas exotoxin (PE) in which the cell binding domain of PE (domain Ia, amino acids 1-252) and a portion of domain Ib (amino acids 365-380) are deleted. The immunotoxins LMB-1 and LMB-7 contain PE38 and kill cancer cells by exploiting the cytotoxic action of PE38. The major human B cell epitopes of PE38 were mapped by measuring the reactivity of 45 serum samples from patients treated with the PE38-containing immunotoxins LMB-1 or LMB 7 to two panels of overlapping synthetic peptides representing the sequence of PE38. One panel of peptides is ten amino acids long and overlap by seven amino acids, and the second panel of peptides is twenty amino acids long and overlap by ten. Five major epitopes were identified: amino acids 274-283, 470-492, 531-540, 555-564, and the C-terminal amino acids 596-609. Two minor epitopes were identified as well: amino acids 501-510 and 582-589. These epitopes are predominantly located on the surface of the protein. The amino acids believed to be critical for binding are highly solvent-accessible residues. The results of the human antibody response to peptides are compared to the pattern of reactivity previously identified with serum samples obtained from monkeys administered LMB-1 and LMB-7. The epitopes between monkey and human are almost identical, demonstrating similarity in the response of antibody repertoires between the two species and providing further support that these are the immunodominant epitopes. This information is critical for genetically engineering less immunogenic immunotoxins and provides a foundation for the development of a vaccine against pseudomonal infections which plague immunocompromised individuals and individuals with cystic fibrosis. PMID- 9209500 TI - Characterization of mouse ALCAM (CD166): the CD6-binding domain is conserved in different homologs and mediates cross-species binding. AB - Activated leukocyte cell adhesion molecule (ALCAM; CD166) is a member of the immunoglobulin gene superfamily (IgSF) which is expressed by activated leukocytes and thymic epithelial cells and is a ligand for the lymphocyte antigen CD6. Herein, we report on the isolation and characterization of cDNA clones encoding mouse ALCAM (mALCAM). Comparison of the predicted amino acid sequence of mALCAM and human ALCAM (hALCAM) showed an overall identity of 93%. Binding studies with truncated forms of the extracellular region of mALCAM showed that the CD6 binding site is located in the N-terminal Ig-like domain and that mALCAM is capable of binding both human and mouse CD6. Mutagenesis studies on hALCAM suggested that residues critical for CD6 binding map to the predicted A'GFCC'C" beta-sheet of ALCAM's N-terminal binding domain. Residue differences in the N-terminal domains of mALCAM and hALCAM were analyzed with the aid of a molecular model of ALCAM. All residues critical for CD6 binding are conserved in both mALCAM and hALCAM, whereas residue differences map to the predicted BED face which is opposite the CD6 binding site on hALCAM. These findings provide a molecular rationale for the observed cross-species CD6/ALCAM interaction and the apparent inability to generate monoclonal antibodies (mAb) against the CD6 binding site. RNA blot analysis showed that mRNA transcripts encoding mALCAM are expressed in the brain, lung, liver, and the kidney, as well as by activated leukocytes and a number of cell lines. A rat mAb specific for mALCAM was produced and by two-color immunofluorescence studies was shown to bind to both activated CD4+ and CD8+ T cells. PMID- 9209501 TI - Late events in the intracellular sorting of major histocompatibility complex class II molecules are regulated by the 80-82 segment of the class II beta chain. AB - The molecular mechanisms that regulate sorting of major histocompatibility complex (MHC) class II molecules into the endocytic pathway are poorly understood. For many proteins, access to endosomal compartments is regulated by cytosolically expressed sequences. We present evidence that a sequence in the lumenal domain of the MHC class II molecule regulates a very late event in class II biogenesis. Class II molecules containing single amino acid changes in the highly conserved 80-82 region of the beta chain were introduced into invariant chain (Ii)-negative fibroblasts with wild-type alpha chain, and the derived transfectants were analyzed biochemically. Using an endosomal isolation technique, we have quantified the level of class II molecules expressed in endocytic compartments and found that in the absence of Ii, approximately 15% of total cellular class II molecules can be isolated from endosomal compartments. Mutation at position 80 enhances this localization, while changes at positions 81 and 82 ablate class II expression in endosomal compartments. In addition, we have evaluated whether the induced changes in intracellular distribution of class II molecules were due to alterations in early biosynthetic events, indicative of misfolding of the molecules, or to modulation of later trafficking events more likely to be a consequence of the modulation of a specific transport event. Despite the dramatic effects on endosomal localization induced by the mutations, early biosynthetic events and maturation of class II were unaffected by the mutations. Collectively, our data argue that late trafficking events that control the ability of the class II molecule to access antigens is regulated by the 80-82 segment of the MHC class II beta chains. PMID- 9209502 TI - Direct cell/cell communication in the lymphoid germinal center: connexin43 gap junctions functionally couple follicular dendritic cells to each other and to B lymphocytes. AB - Direct cell/cell communication occurs through gap junctions (GJ). We mapped GJ expression in secondary lymphoid organs and found, for the first time, a high density of connexin43 (Cx43) GJ in follicular dendritic cells (FDC) in close association with lymphocytes (Krenacs T. and Rosendaal M., J. Histochem. Cytochem. 1995. 43: 1125-1137). In this work, we used a combination of ultrastructural, immunocytochemical, molecular methods, and functional dye transfer experiments to study which germinal center cells are involved in direct cell/ cell communication and how GJ expression is regulated during antigen responses. One week after injecting the footpad of mice with 50 micrograms lysozyme, Cx43 GJ were detected on elongated cells in the paracortex of their popliteal lymph nodes. Repeated challenge led to the formation of secondary follicles with enlarged FDC meshwork full of Cx43 GJ. This positive correlation may reflect an importance for GJ in the pattern formation of FDC and lymphoid follicles. In human tonsil, the density of GJ and FDC was highest in the light zone of germinal centers where the fate of B cells is thought to be decided. Cx43 colocalized with CD21 and CD35 antigens in the vicinity of desmosomal junctions on FDC embracing lymphocytes. Freeze-fracture hallmarks of GJ of 200-400 nm were also found on FDC in the vicinity of desmosomal plaques. Furthermore, Northern blot analysis showed the consistent presence of Cx43 mRNA in human tonsil and spleen. Most Cx43 message was localized in situ to cells with FDC morphology and some to a few germinal center lymphocytes. To investigate functional cell coupling, we set up FDC/B cell cultures from the low density cell fractions of human tonsils. Cx43 plaques associated with lymphocytes were detected both on elongated FDC processes in early cultures (up to 4 h) and in established FDC/B cell clusters (between 4 and 24 h). In early cultures, we injected FDC with Lucifer Yellow, a fluorescent dye which passes through GJ: the dye spread into adjacent FDC and occasionally from FDC into CD19+ B cells. Based on these results, we propose that direct cell/cell communication through Cx43 GJ is involved in FDC/FDC and in FDC/B cell interactions. The functionally coupled FDC meshwork may serve as a communication channel synchronizing germinal center events. FDC may also deliver crucial direct signals through GJ involved in the rescue of high-affinity B cell clones from apoptotic cell death. PMID- 9209503 TI - The role of E- and P-selectin in neutrophil and monocyte migration in adjuvant induced arthritis in the rat. AB - The role of the endothelial adhesion molecules E- and P-selectin in leukocyte accumulation in arthritis is not known. We investigated this role in rat adjuvant arthritis by employing adhesion function-blocking monoclonal antibodies (mAb) to rat P- and E-selectin. The acute migration (2 h) of radiolabeled rat blood neutrophils and monocytes to joints and skin was determined. Anti-P-selectin mAb significantly reduced accumulation of monocytes (by 50%) and neutrophils (by 40%) in the talar joint, and of neutrophils in tail joints (by 90%). Anti-E-selectin mAb alone did not attenuate leukocyte migration, but when combined with anti-P selectin mAb, it enhanced inhibition of neutrophil accumulation in the talar and carpal joints. In the same animals, anti-P-selectin mAb significantly inhibited neutrophil and monocyte migration to dermal inflammatory reactions induced by zymosan-activated rat serum (ZAS) containing the chemotactic factor C5ades Arg, endotoxin (LPS), interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha). In contrast, anti-E-selectin mAb alone had no effect on monocyte or neutrophil accumulation in inflamed skin of arthritic animals, but again enhanced the inhibition when combined with mAb to P-selectin. The addition of anti-L selectin mAb to anti-P- and E-selectin mAb did not further suppress monocyte or neutrophil migration to inflamed skin or joints. These results demonstrate that optimal leukocyte migration to arthritic joints and inflamed skin is P-selectin dependent, and E-selectin is not essential. However, E-selectin contributes to migration when P-selectin mechanisms are not operative. L-selectin does not play a role in E- and P-selectin-independent leukocyte migration to joints or skin inflammation in arthritic rats. However, it is likely that additional selectin independent pathways also mediate neutrophil and monocyte migration to joint and skin inflammation. PMID- 9209504 TI - Excessive degradation of intracellular protein in macrophages prevents presentation in the context of major histocompatibility complex class II molecules. AB - The endogenous major histocompatibility complex (MHC) class II presentation pathway allows biosynthesized, intracellular antigens access for presentation to MHC class II-restricted T cells. This pathway has been well documented in B cells and fibroblasts, but may not be universally available in all antigen-presenting cell types. This study compares the ability of different antigen-presenting cells, expressing endogenous C5 protein (fifth component of mouse complement) as a result of transfection, to present their biosynthesized C5 to MHC class II restricted T cells. B cells and fibroblasts expressing C5 were able to present several epitopes of this protein with MHC class II molecules, whereas macrophages were unable to do so, but readily presented C5 from an extracellular source. However, macrophage presentation of endogenous C5 could be achieved when they were treated with low doses of the lysosomotropic agent ammonium chloride. In the presence of an inhibitor of autophagy, presentation of endogenous C5 was abrogated, indicating that biosynthesized C5 is shuttled into lysosomal compartments for degradation before making contact with MHC class II molecules. Taken together, this suggests that proteolytic activity in lysosomes of macrophages may be excessive, compared with fibroblasts and B cells, and destroys epitopes of the C5 protein before they can gain access to MHC class II molecules. Thus, there are inherent differences in presentation pathways between antigen presenting cell types; this could reflect their specialized functions within the immune system with macrophages focussing preferentially on internalization, degradation, and presentation of extracellular material. PMID- 9209505 TI - Combined structural and immunological refinement of HIV-1 HLA-B8-restricted cytotoxic T lymphocyte epitopes. AB - This study demonstrates that use of structural information improves the definition and optimization of cytotoxic T lymphocyte (CTL) epitopes. Epitope optimization usually requires numerous truncated peptides or a reverse immunogenetic approach, where the peptide binding motif is used to predict epitopes. These binding motifs do not reliably predict all peptides which are CTL epitopes. Comparison of 24 peptides eluted from HLA-B8 with 10 HLA-B8-restricted defined CTL epitopes demonstrated that known epitopes varied considerably at anchor positions. We used structural information based on determination of the crystal structure of the HLA-B8-GGKKKYKL complex to reassess previously described CTL epitopes, to predict new epitopes, and to predict the consequences of naturally occurring variation within epitopes. These predictions were confirmed by cytotoxicity and binding assays. Use of combined structural and immunological data more accurately defines the true peptide-binding motif of a restriction element than eluted peptide data allows. PMID- 9209506 TI - The C terminus of the human C5a receptor (CD88) is required for normal ligand dependent receptor internalization. AB - The biological effects of the potent inflammatory mediator C5a, a complement split product, on human neutrophils and monocytes are limited by the rapid internalization of its specific receptor (C5aR, CD88). The C terminus of the C5aR is phosphorylated after stimulation with C5a of phorbol ester, and this phosphorylation might lead to receptor internalization. In this context, we have studied the effects on C5aR internalization of C5a, phorbol 12-myristate 13 acetate (PMA), the protein kinase inhibitor staurosporine, and pertussis toxin on rat basophilic RBL.2H3 cells stably transfected with the human wild-type or mutant C5aR. C5aR mutants lacked either part of the cytosolic C terminus, including suggested major phosphorylation sites, or a putative phosphorylation motif for protein kinase C in the third cytosolic loop. Additionally, agonist induced internalization was analyzed on HEK293 cells co-transfected with C5aR and the pertussis toxin-resistant G protein alpha subunit, G alpha 16. Staurosporine sensitive agonist-dependent C5aR internalization could be detected, suggesting that C5aR phosphorylation, most likely of the C terminus, participates in this type of internalization. In contrast, PMA-induced C5aR internalization seems to be independent of putative phosphorylation sites in either the truncated section of the C terminus or the third cytosolic loop. The phorbol ester-induced C5aR internalization may, therefore, be caused by an indirect and less specific effect of protein kinase C on the internalization machinery. Manipulation of the pertussis toxin-sensitive or -resistant G protein-dependent signal transduction had no effect on ligand-induced internalization. PMID- 9209507 TI - Clustering the adhesion molecules VLA-4 (CD49d/CD29) in Jurkat T cells or VCAM-1 (CD106) in endothelial (ECV 304) cells activates the phosphoinositide pathway and triggers Ca2+ mobilization. AB - Ligation of very late antigen (VLA)-4 (alpha 4 beta 1 integrin) with a cross linked anti-alpha 4 subunit monoclonal antibody (mAb) triggered a biphasic Ca2+ response in Jurkat cell populations and in peripheral human lymphocytes. Cross linking vascular cell adhesion molecule (VCAM)-1 (the counter-receptor of VLA-4) in ECV 304 endothelial cells generated a biphasic Ca2+ response. Tumor necrosis factor-alpha-primed human umbilical cord vascular endothelial cells also responded to the cross-linked mAb with a biphasic Ca2+ profile. Ligated VLA-4 (Jurkat cells) or VCAM-1 (ECV 304) stimulated the production of myo-inositol 1,4,5-trisphosphate. ECV 304 cells induced a biphasic Ca2+ response in Fura2 loaded Jurkat cells, whereas a transient response was observed when Jurkat cells were added to Fura2-loaded ECV 304 cells. The Ca2+ responses in these experiments involved VLA-4/VCAM-1 interactions since they were significantly reduced (approximately 80%) by prior treatment of the target cells with the relevant noncross-linked mAb. Close contact between the cells triggered mutual Ca2+ signaling as shown by spectrofluorimetric and confocal microscopy time-dependent recordings. Fibronectin and its CS-1 fragment (V25) triggered a sustained Ca2+ response in Jurkat cells (confocal microscopy). Our results suggest that the VLA 4 and VCAM-1 adhesion molecules can transduce a signal that involves activation of the phosphoinositide pathway and the mobilization of Ca2+. PMID- 9209509 TI - Differential expression of insulin-dependent diabetes mellitus-associated HLA DQA1 alleles in vivo. AB - The strong association of HLA-DQ genes with insulin-dependent diabetes mellitus (IDDM) susceptibility is persuasive evidence of their central role in the etiology of this autoimmune disease. Among other possibilities, it has been proposed that an unbalanced expression of IDDM-associated DQA, and/or DQB alleles may lead to alterations in the composition of alpha beta heterodimers and preferential expression of a particular heterodimer on the antigen-presenting cell surface, leading to self-recognition. In this report, we demonstrate the differential expression of DQA1 alleles in vivo, in particular of the two diabetogenic alleles DQA1*0301 and DQA1*0501. Family studies suggest that unequal HLA-DQA1 allele expression in heterozygous individuals is not associated in cis with the HLA-DQA1 gene, but may be affected by trans-acting determinant(s). We also discuss the segregation of this phenotype in IDDM-affected members. Furthermore, we examined historical samples of PBL from an IDDM-affected individual and an HLA-identical unaffected sibling acting in a kidney transplant program as donor and recipient, respectively. This analysis allowed us to establish that unbalanced expression of DQA1*0301 and DQA1*0501 can be induced by microenvironmental conditions. Inducible differential expression of HLA-DQA1 alleles may account for the discordance in the outcome of autoimmune disease in monozygotic twins and HLA-identical siblings. PMID- 9209508 TI - Cytokine responses induced by Toxoplasma gondii in astrocytes and microglial cells. AB - To investigate the role of astroglia in intracerebral immune response to Toxoplasma gondii, astrocytes cultured from mouse brain were inoculated with mouse-virulent or -avirulent toxoplasma strains. In comparison to microglia/ brain macrophages, astrocytes as host cells allowed stronger proliferation of avirulent parasites. Toxoplasma infection of astroglia was accompanied by release of interleukin- (IL)1 alpha, IL-6, and granulocyte/macrophage colony-stimulating factor (GM-CSF) activity, whereas alternative challenge by lipopolysaccharide (LPS) evoked no IL-1 response and significantly higher titers of IL-6 and GM-CSF. At the mRNA level, both stimuli induced transcription of all three cytokines in astrocytes. Secretion of IL-1 and IL-6 upon infection was triggered by T. gondii brady- and tachyzoites in a time- and dose-dependent manner. Heat killing of parasites, but not an exposure to polymyxin B, abrogated their cytokine-inducing activity, thus indicating that an LPS-independent stimulus is provided by T. gondii. When administered in combination, LPS synergistically augmented the IL-1 inducing effect of toxoplasma infection. In comparison, T. gondii-induced, but not an LPS-triggered, IL-6 response of astrocytes resisted to antagonization with IL-10. The IL-6 response of parasitized astroglia was up-regulated by external tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta 1, with only TNF-alpha enhancing simultaneous release of IL-1. Substantial secretion of IL-10 and TNF-alpha was detected in T. gondii-infected microglia, but not in astrocyte cultures. A possibly autocrine stimulation of infected astroglia via IL 1 was found to be unlikely, since addition of IL-1 receptor antagonist did not affect the release of IL-6 and GM-CSF while inhibiting these responses in IL-1 treated cells. The findings substantiate a separate, T. gondii-induced pathway of astroglia activation characterized by the release of IL-1 which may drive local inflammatory reaction both at initial infection of the brain and during reactivating toxoplasmosis. PMID- 9209510 TI - The repertoire of serum IgM in normal mice is largely independent of external antigenic contact. AB - Antigen-free (AGF) and germ-free (GF) mice, although essentially free of serum IgG, maintain normal levels of circulating IgM. Using a quantitative immunoblot assay, we have now analyzed the repertoire of serum IgM from AGF, GF, and specific pathogen-free (SPF) BALB/c mice, on large panels of natural antigens from homologous tissues and bacteria. The reactivity profiles were very similar in the three groups of mice. Multiparametric statistic evaluation of the data showed that BALB/c animals, SPF, GF, and AGF mice constitute an homogeneous group with similar immunoreactivity profiles when compared to C57BL/6. Differences between immunoreactivity profiles of GF and AGF mice were observed, but were not statistically significant. These results suggest that the serum IgM repertoire of normal mice is strictly regulated and selected by endogenous ligands. PMID- 9209511 TI - Antigen specificity of anti-nuclear antibodies complexed to nucleosomes determines glomerular basement membrane binding in vivo. AB - Monoclonal anti-nuclear antibodies which are complexed to nucleosomes are able to bind to the glomerular basement membrane (GBM) in vivo, whereas purified antibodies do not bind. The positively charged histone moieties in the nucleosome are-responsible for the binding to anionic determinants in the GBM. We tested the hypothesis that the specificity of the autoantibodies complexed to the nucleosome influences the glomerular binding of the antibody-nucleosome complex. We induced the formation of these immune complexes in vivo, by intraperitoneal inoculation of hybridomas producing monoclonal anti-nuclear antibodies (four anti-histone, three anti-double stranded (ds)DNA and three anti-nucleosome antibodies) into nude BALB/c mice. In ascites and plasma from the mice inoculated with these hybridomas, nucleosome/autoantibody complexes were detected in comparable amounts. Immunofluorescence of kidney sections revealed that about 60% of the mice inoculated with anti-nucleosome or anti-dsDNA hybridomas had immunoglobulin deposits in the GBM, whereas only 15% of the mice with anti-histone hybridomas showed these deposits (p < or = 0.04). In the Matrigel-ELISA (used as a GBM surrogate) ascites from anti-nucleosome or anti-DNA hybridomas displayed significantly higher titers (p < or = 0.002) than ascites from anti-histone hybridomas. In conclusion, nucleosome/immunoglobulin complexes comprising anti nucleosome or anti-dsDNA auto-antibodies do bind more frequently to the GBM in vivo than nucleosome/immunoglobulin complexes containing anti-histone antibodies. It therefore appears that the specificity of the antibody bound to the nucleosome is a critical determinant for the nephritogenic potential of the nucleosome autoantibody complex. PMID- 9209512 TI - The defined attenuated Listeria monocytogenes delta mp12 mutant is an effective oral vaccine carrier to trigger a long-lasting immune response against a mouse fibrosarcoma. AB - Listeria monocytogenes has been proposed as a carrier to elicit major histocompatibility complex class-I restricted immune responses able to protect against tumor challenge. In this study the properties of the attenuated L. monocytogenes delta mp12 mutant has been evaluated in vivo against a highly aggressive mouse fibrosarcoma which expresses beta-galactosidase (beta-gal) as a tumor-associated antigen (TAA). Immunization with the vaccine prototypes resulted in both elicitation of specific antibodies and generation of cytotoxic lymphocytes (CTL). Oral vaccination protected 55-64% of the immunized animals from tumor take (p < 0.01) and strongly reduced the average size of the tumor in the other 34-45% (p < 0.01). Vaccinated mice developed a long-lasting response, which resulted in 100% protection from a subsequent tumor challenge. Substitution of the whole TAA by its CTL-defined immunodominant epitope resulted in 43% protection, suggesting a contribution of the humoral response to the observed antitumor effect. No statistically significant differences were observed in the antitumor response when mice were immunized with strains expressing the immunodominant TAA epitope in the context of carrier proteins which were either exported or restricted to the bacterial cytoplasm. This suggests that the topology of the recombinant antigen does not play a major role in the outcome of the protective response. PMID- 9209514 TI - Prevention of endotoxin-induced lethality in neonatal mice by interleukin-13. AB - Interleukin(IL)-13, a cytokine produced by T helper 2 (Th2) cells, is a powerful inhibitor of macrophage functions, including surface expression of CD14 and production of IL-1 and tumor necrosis factor (TNF)-alpha. We tested the effects of recombinant mouse(m)IL-13 in a neonatal mouse model of endotoxin shock; this is a macrophage-dependent condition, which is a model of neonatal sepsis in humans. mIL-13 (0.5 microgram/mouse) dramatically reduced the lethal effects of lipopolysaccharide (LPS) if administered either 24 or 4 h prior to or concomitantly with LPS challenge. This action might be mediated by multiple modulatory activities of IL-13 on LPS-induced cytokine secretion since, relative to control animals, the mice treated with mIL-13 had eight times lower peak blood levels of TNF. The IL-1 beta levels were also decreased, whereas increased levels of IL-6 and IL-10 were observed at several time points after LPS challenge. PMID- 9209513 TI - Lack of directed V alpha 14-J alpha 281 rearrangements in NK1+ T cells. AB - NK1.1+ T cells are an unusual subset of TCR alpha beta cells distinguished by their highly restricted V beta repertoire and predominant usage of an invariant V alpha 14-J alpha 281 chain. To assess whether a directed rearrangement mechanism could be responsible for this invariant alpha chain, we have analyzed V alpha 14 rearrangements by polymerase chain reaction and Southern blot in a panel of cloned T-T hybrids derived from thymic NK1.1+ T cells. As expected a high proportion (17/20) of the hybrids had rearranged V alpha 14 to J alpha 281. However, V alpha 14-J alpha 281 rearrangements always occurred on only one chromosome and were accompanied by other V alpha-J alpha rearrangements (not involving V alpha 14) on the homologous chromosome. These data argue that rigorous ligand selection rather than directed rearrangement is responsible for the high frequency of V alpha 14-J alpha 281 rearrangements in NK1.1+ T cells. PMID- 9209515 TI - Major histocompatibility complex-controlled protective influences on experimental autoimmune encephalomyelitis are peptide specific. AB - The myelin basic protein (MBP) peptide 63-88-induced experimental autoimmune encephalomyelitis (EAE) and its associated T cell cytokine profile are influenced by the rat major histocompatibility complex (MHC). There is an allele-specific protective influence of the MHC class I region, whereas the MHC class II region display either disease-protective or -promoting effects. To investigate if the MHC-associated protection is dependent on certain combinations of MBP peptide and MHC molecules, we have now used another peptide (MBP 89-101). A broader and different set of rat MHC alleles were associated with EAE induced with MBP 89-101 as compared to MBP 63-88. All EAE-susceptible strains mounted peptide-specific strong T helper (Th) 1-like immune responses in vitro. Immunization of rats with an extended peptide (MBP 87-110) induced EAE associated with the same MHC haplotypes as the 89-101 peptide, except in LEW.1N (RT1 pi) rats which were relatively resistant. Only this strain responded with additional Th2-like and transforming growth factor-beta responses to the peptide in vitro. In vivo depletion of CD8+ cells aggravated the disease in this strain. We conclude that both MHC-controlled promoting and protective influences on EAE are dependent on certain MHC/MBP peptide combinations, and that the 87-110 region of MBP contains a major MHC-associated encephalitogenic epitope in the rat. PMID- 9209516 TI - 5-year cure rate: yet another myth. PMID- 9209517 TI - Results of preoperative intraluminal brachytherapy combined with radical surgery for middle and lower rectal carcinomas. AB - BACKGROUND: Radiation therapy in the treatment of rectal carcinoma has received attention. We attempted to learn whether preoperative intraluminal brachytherapy (IBT) gives an advantage in local control and/or prolongation of survival. METHODS: One hundred and fifteen patients with middle and lower rectal carcinoma with penetration into or through the rectal wall were consecutively treated with preoperative IBT and radical operation. Patients were divided into the moderate dose group (group A: 16-40 Gy; n = 96) and the high-dose group (group B: 40-80 Gy; n = 19). A control group of 115 rectal carcinoma patients who received no radiation prior to radical surgery was compared (group C). RESULTS: The rate of sphincter-saving resection was 72% in group A, 63% in group B, and 42% in group C (group A vs. group C; P < 0.0001). The local recurrence rate at 5 years was 11% in group A, 6% in group B, and 26% in group C (group A vs. group C; P = 0.005). The 5-year survival rate was similar among the three groups. CONCLUSIONS: These results suggested that IBT contributed to the improvement of local control but not survival after radical resection of rectal carcinomas. The application of IBT might be useful in preserving the intestinal continuity for rectal carcinomas. PMID- 9209518 TI - A clinical and pathologic study on para-aortic lymph node metastasis in endometrial carcinoma. AB - BACKGROUND: Recent studies have shown that poor survival for patients with early endometrial cancer was related to the extrapelvic spread of the cancer. The purpose of this study was to evaluate the correlation between para-aortic lymph node (PAN) metastasis and histopathologic findings and to assess the clinical utility of identifying PAN metastasis of endometrial carcinoma. METHODS: The correlation of para-aortic lymph node metastasis to the clinical stages of endometrial carcinoma (FIGO, 1982), histopathologic findings, and prognosis were investigated in 200 patients with endometrial carcinoma, who were treated by radical operations, including systematic retroperitoneal lymphadenectomies, between July 1982 and February 1996. RESULTS: Of these, para-aortic lymph node (PAN) metastasis was seen in 18 (9.0%) and pelvic lymph node (PLN) metastasis in 40 (20.0%). The incidence of PAN metastasis according to clinical stages Ia, Ib, II, and III were 2.5%, 8.5%, 15.7%, and 33.3%, respectively. The incidence of metastasis was significantly higher in stage II than in stage Ia (P < 0.05), and in stage III than in stage Ia (P < 0.01). PAN metastasis occurred significantly more frequently in the first of each of the following groups: invasion of > 1/2 of the myometrium (15.7%) vs. invasion of < 1/2 of the myometrium (3.6%) (P < 0.01), the group with cervical invasion (23.5%) vs. the group without (4.0%) (P < 0.0001), the group with lymph-vascular space involvement (17.2%) vs. the group without (1.0%) (P < 0.0005), and PLN-metastasis-positive group (40.0%) vs. the negative group (1.3%) (P < 0.0001). Multivariate analysis showed a significant correlation between PAN and PLN metastases (P < 0.0005). Positive PAN metastasis is not related to multiple PLN metastasis (bilateral PLN metastasis and the number of PLN metastatic groups). However, a correlation was seen between PAN metastasis and common iliac node metastasis. The prognosis was significantly poorer (P < 0.05) for patients with both PLN and PAN metastases than for those with PLN metastasis alone. CONCLUSIONS: The results of the present study suggest that PAN metastasis may occur as a consequence of PLN metastasis or the two may occur simultaneously as PLN metastasis and also that careful examination of PAN metastasis is necessary to determine the prognosis. PMID- 9209519 TI - Complications of combined radiotherapy and isolated limb perfusion with tumor necrosis factor alpha +/- interferon gamma and melphalan in patients with irresectable soft tissue tumors. AB - BACKGROUND: Isolated limb perfusion (ILP) with tumor necrosis factor alpha (TNF alpha) +/-interferon gamma (IFN gamma) and melphalan in patients with primarily irresectable soft tissue sarcoma is promising in terms of tumor regression and limb salvage. However, the feasibility of radiotherapy in combination with this treatment modality has not been established. METHODS: Fifteen patients with irresectable soft tissue tumors of the limb underwent ILP with TNF alpha, +/-IFN gamma, and melphalan. Three groups could be distinguished with respect to the role of radiotherapy. In nine patients, the residual tumor could be resected after ILP, and this was followed by radiotherapy with a total dose of 50-70 Gy (2 Gy/day). In one patient with aggressive fibromatosis, ILP was followed by radiotherapy without tumor resection (Group I). In two patients who underwent ILP for recurrent sarcoma, the primary tumor had been treated before by resection and radiotherapy (60 Gy) (Group II). In three patients whose tumors remained irresectable after ILP, radiotherapy was applied later in the course of disease for local palliation (Group III). RESULTS: In Group I, healing of the resection wound was markedly delayed in four patients, with soft tissue necrosis and infection necessitating amputation in two of them. Following completion of radiotherapy, persistent lymphoceles were encountered in two patients. Radiotherapy-induced fibrosis was encountered in five patients, resulting in a mild limb malfunction in two. Three-patients developed mild edema during radiotherapy. Tumor-associated neuropathy was aggravated by ILP in three patients causing severely disabling motor deficits and limb contractures in two of them. In Group II, ILP did not cause any local problem in the heavily irradiated areas. In Group III, pre-existing limb edema was increased after a total palliative dose of 20 Gy in one patient. Another patient, who had been re-operated for arterial thrombosis immediately after ILP, developed occlusion of the brachial artery 4 months after completion of palliative radiotherapy (36 Gy in 6 Gy fractions). CONCLUSIONS: In patients with irresectable soft tissue tumors, multimodality treatment using ILP with TNF alpha +/- IFN gamma and melphalan, tumor resection, and postoperative high-dose radiotherapy is associated with a considerable risk of tissue necrosis and impaired healing. This risk should be weighed against a possible benefit from radiotherapy in local tumor control. PMID- 9209521 TI - Effect of preoperative 5-fluorouracil on apoptosis of advanced gastric cancer. AB - BACKGROUND: Several studies have reported on apoptosis and the effect of anticancer chemotherapy. METHODS: We studied apoptosis induced by 5-fluorouracil (5-FU) given preoperatively to 28 patients with advanced gastric cancer and compared the findings with 101 untreated patients. The expression of bcl-2 oncoprotein, cell phase fractions, and histological chemotherapeutic effects were also compared with the apoptotic changes. RESULTS: The apoptotic and S-phase fractions in 5-FU-treated patients (apoptotic fraction: 10.46 +/- 6.93%, S-phase fraction: 17.49 +/- 11.65%) were significantly greater than those in untreated controls (apoptotic fraction: 6.56 +/- 5.06%, S-phase fraction: 12.17 +/- 6.78%). A positive correlation was observed between 5-FU-induced apoptosis and accumulation of tumor cells in the S-phase fraction. There was an inverse relationship between bcl-2 oncoprotein expression and apoptosis in 5-FU-treated patients, but no significant correlation between histological effect and apoptosis. However, two patients with significant histological effects showed no bcl-2 oncoprotein expression, whereas the histological effects were mild in all the bcl-2-positive patients. CONCLUSIONS: Apoptosis may be induced by 5-FU administered preoperatively and bcl-2 oncogene expression may suppress 5-FU induced apoptosis. PMID- 9209520 TI - P-glycoprotein expression in cartilaginous tumors. AB - BACKGROUND AND OBJECTIVES: Malignant cartilage tumors demonstrate chemotherapeutic resistance through undetermined mechanisms. P-glycoprotein is the protein product of the multiple drug resistance gene 1 (MDR-1) and confers multidrug chemotherapeutic resistance in a variety of malignancies. METHODS: MDR 1 expression was examined in 55 benign and malignant cartilage tumor specimens by immunohistochemistry using C219, C494, and JSB-1 antibodies, and by in situ hybridization with an MDR-1 specific oligonucleotide cDNA probe. RESULTS: Constitutive expression of P-glycoprotein was observed in all benign and malignant cartilage tumor specimens with a similar pattern of immunohistochemical staining present with all three antibodies. In benign tumors and low grade chondrosarcomas, the staining pattern was weak to intermediate and localized to clusters of cells. However, higher grade-tumors (Grade II and III) expressed P glycoprotein in a higher percentage of cells and with more intense staining. P glycoprotein expression was absent in normal human articular cartilage, but was focally present in costal and growth plate cartilage. The immunohistochemistry results were confirmed by in situ hybridization in 10 cases. CONCLUSIONS: P glycoprotein is expressed constitutively in cartilaginous tumors, with greatest expression in high grade malignancies. The findings may account for the resistance of cartilage tumors to chemotherapeutic agents. PMID- 9209522 TI - Clinical significance of p53, nm23, and bcl-2 in T3-4N1M0 oesophageal carcinoma: an immunohistochemical approach. AB - BACKGROUND AND OBJECTIVE: The purpose of this retrospective study was to test whether the expression of p53, nm23, and bcl-2 in T3-4N1M0 oesophageal carcinoma is associated with patient survival. METHODS: Immunohistochemical localisation of p53, nm23, and bcl-2 was performed on formalin-fixed, paraffin-embedded tissue sections (N = 46). The observed range of follow-up period was 0.2-24.0 months with a median of 11.0 months. A total of 85% (39/46) of the patients died within 24.0 months, which could be due to advanced disease at presentation. The immunohistochemical signal was expressed as the proportion of positive cells. The immunostaining for p53 was nuclear, whereas that for nm23 and bcl-2 was cytoplasmic in the neoplastic cells. RESULTS: p53 was expressed in 70% (32/46) of cases; nm23 in 29% (13/45), and bcl-2 in 67% (29/43) of tumours. The univariate analysis showed that the expression of two markers, i.e., expression of p53 and absence of nm23 were independently associated with unfavourable overall survival time. Despite a small number of patients treated with adjuvant therapy, we observed that tumours positive for p53 had an unfavourable prognosis when compared with tumours negative for p53. CONCLUSIONS: Our preliminary findings suggest: expression of p53 and nm23 negativity may be related to an unfavourable prognosis in patients with advanced oesophageal carcinoma. PMID- 9209524 TI - Experimental pulmonary sarcoma metastases in athymic nude mice. AB - BACKGROUND: Pulmonary metastases remain a challenging therapeutic problem in the treatment of patients with soft tissue sarcomas. A pulmonary sarcoma metastases animal model might facilitate studying the biology of metastases, diagnosis, and treatment modalities of this disease. Intravenous injection of human tumor cells into nude mice has been reported using human melanoma and colorectal carcinoma to produce pulmonary metastases. Human fibrosarcoma cells were intravenously administered to athymic nude mice to simulate clinical pulmonary metastases. METHODS: HT-1080 human sarcoma cells derived from a poorly differentiated fibrosarcoma were used to prepare inoculant at a concentration of 5 x 10(6) cells per ml. Male athymic nude mice were injected subcutaneously with 1 x 10(6) cells in the right hind flank and sacrificed when the tumors were 1-2 cm in diameter. Age- and weight-matched athymic nude mice were intravenously injected through tail veins with 10(4), 10(5), and 10(6) cells. The mice were sacrificed at 7, 14, and 21 days after intravenous injection of the tumor cells. Tissues were histologically examined for pulmonary metastases. RESULTS: Neither gross nor microscopic spontaneous metastases were found in any of the animals that received subcutaneous xenografts, and no pulmonary metastases were identified in mice intravenously injected with < 10(5). All mice inoculated with 10(6) cells developed tumor colonies in the lungs, which were microscopically evident as early as day 7. No metastases were found in the liver, spleen, heart, or other tissues. In a second experiment, HT-1080 cells were injected at 10(6); all animals developed lung metastases and died of lung tumor involvement, with an average survival of 35 days. CONCLUSIONS: These experiments identify a sarcoma animal pulmonary metastases model that is readily available, relatively inexpensive, easily utilized, and reproducible. PMID- 9209523 TI - Prognostic factors for fibromatoses: a correlation of proliferation index, estrogen receptor, p53, retinoblastoma, and src gene products and clinical features with outcome. AB - BACKGROUND: The aggressiveness of fibromatoses is difficult to predict by morphologic analysis. Additional prognostic markers would be helpful for clinical management. MATERIALS AND METHODS: Proliferation index (MIB-1), p53, src, retinoblastoma gene protein products, estrogen receptor level, site and depth of lesion were correlated with incidence of recurrence in 52 patients. Superficial (47) and deep (5) fibromatoses were studied. Anatomic sites included the extremities, head, neck, trunk, and pelvis. RESULTS: Twenty (38%) lesions recurred locally. All five deep lesions recurred, but only 32% of superficial tumors recurred. Mean proliferation index for recurrent lesions was 0.82% and 0.73% for nonrecurrent fibromatoses; no significant differences were observed. Five recurrent lesions (25%) expressed estrogen receptor > 5 fmol/mg as did 31% (10 of 32) of the nonrecurrent lesions. None of the tested specimens expressed src gene product. Eight of the lesions which recurred (40%) contained p53, but only five nonrecurring tumors (16%) expressed p53. One of five deep lesions (20%) expressed p53 and 26% (12 of 47) of superficial tumors expressed p53. Forty-six percent (6 of 13) of recurrent lesions tested were retinoblastoma protein product negative, but only 33.3% (7 of 21) of nonrecurring tumors were retinoblastoma protein product negative. CONCLUSIONS: Only p53 and depth of lesion were of statistical value for the prediction of recurrence. PMID- 9209525 TI - Superselective intra-arterial chemotherapy with mitomycin C in hepatic metastases from colorectal cancer. AB - BACKGROUND: Mitomycin C has been found clinically useful in the treatment of colorectal cancer when administered via the hepatic artery. In a prospective therapeutic trial, we studied the effect of superselective intra-arterial chemotherapy with mitomycin C in patients with hepatic metastases from colorectal cancer. METHODS: Forty-six patients with hepatic metastases from colorectal cancer received intra-arterial chemotherapy with mitomycin C (SIAC) between 1981 and 1991. The results of a 5-year follow-up were compared with 46 control patients standardized by sex, age, and tumor distribution. RESULTS: The overall response rate to intra-arterial chemotherapy was 20%. The median survival time for responders was 26 months and that for nonresponders 12 months (P < 0.003). The median survival period after intra-arterial chemotherapy was 15 months, compared with 9 months in controls (P < 0.004). The cumulative 5-year survival rate was 6% for patients treated by SIAC and 5% for controls. Cessation of chemotherapy was necessary in 39 of the 46 patients: in 28 because of tumor progression, in 9 because of toxicity, in 1 because of catheterization difficulties, and in 1 because of patient refusal. CONCLUSIONS: Superselective intra-arterial mitomycin C therapy had a poor effect on hepatic metastases from colorectal cancer because of the low response and long-term survival rates. PMID- 9209526 TI - Reliability of frozen section diagnosis of gallbladder tumor for detecting carcinoma and depth of its invasion. AB - BACKGROUND: An accurate frozen section diagnosis is important when deciding the surgical strategy against a gallbladder tumor intraoperatively. Little has been reported on the accuracy of frozen section diagnosis of the gallbladder. PATIENTS AND METHODS: In a total of 86 consecutive patients with gallbladder tumor, the accuracy of the frozen section diagnosis was examined. There were 32 patients with polypoid lesions and 54 with nonpolypoid tumors. RESULTS: The frozen tissue diagnosis and final diagnosis were identical in 82 of the 86 cases, that is, benign in 65 and malignant in 17. The other four cases had different diagnoses, that is, conversion from benign to malignant in two and from malignant to benign in two. The overall accuracy of frozen diagnosis was 95.3% (97.0% for benign and 94.7% for malignant). In 32 polypoid lesions, the accuracy of frozen section diagnosis was 91% (93% for benign; 89% for malignant). In 54 nonpolypoid lesions, the accuracy of diagnosis was 98% (100% for benign; 93% for malignant). The diagnosis of depth of invasion was identical only in 7 (70%) of the 10 carcinoma cases examined, while it was diverse in the remaining 3, that is, conversion from adenocarcinoma invading the subserosa to that limiting to the mucosa in one, from carcinoma within the mucosa to that infiltrating the muscle coat in one, and from carcinoma affecting the muscle layer to that invading the subserosa in the other. Alterations of frozen section diagnosis about being benign or malignant and about the depth of invasion were encountered in seven patients, five of whom had a polypoid tumor. CONCLUSIONS: The intraoperative frozen tissue diagnosis is fairly reliable as to whether lesions are malignant or benign; however, accuracy is low in patients with polypoid lesions of the gallbladder. Also, frozen section diagnosis does not reliably measure the depth of invasion of gallbladder carcinoma. PMID- 9209527 TI - Thyroid hemiagenesis. AB - BACKGROUND AND OBJECTIVES: Thyroid hemiagenesis is a rare embryological condition, predominantly in females (3:1) with a left lobe being absent. The associated diseases in the remaining thyroid lobe include benign adenoma, multinodular goiter, hyperthyroidism, chronic thyroiditis, and rarely carcinoma. The most common pathology involved in thyroid hemiagenesis is hyperthyroidism. Presence of carcinoma in a patient with hemiagenesis is quite rare and very few cases are reported in the world literature. METHODS: We report a 30-year-old female who presented with left thyroid mass gradually increasing in size over a period of 3 months. The patient's pre-operative workup included a thyroid scan, which revealed a cold nodule in the left lobe with absent right lobe. A fine needle aspiration biopsy was suspicious for papillary thyroid carcinoma. The patient underwent thyroid exploration and left thyroid lobectomy. RESULTS: The operative findings confirmed hemiagenesis of the right lobe and papillary carcinoma in the left lobe. All four parathyroids were in normal position. CONCLUSIONS: The purpose of this presentation is to discuss and review the literature on thyroid hemiagenesis and present a rare case of absent right thyroid lobe with carcinoma in the remaining left thyroid lobe. PMID- 9209528 TI - Laparoscopic maneuvers in the presence of ascites. PMID- 9209529 TI - Physics and basic parameters of brachytherapy. AB - Brachytherapy (short-distance therapy) is the therapeutic process whereby radioactive sources are placed into very close proximity to target tissue. Radioactive materials were so used beginning shortly after the discovery of radium by Marie and Pierre Curie in 1898. For the purposes of brachytherapy, radioactive materials are those that emit "rays" that can cause ionization (and hence DNA damage and the destruction of target cells). The potentially useful rays include beta, gamma, and other possibilities such as neutrons. Beta rays, properly beta particles, are simply high energy electrons. Gamma rays are high energy photons (part of the electromagnetic spectrum like visible light, but with much higher energy). These particles are produced during the radioactive decay of certain isotopes. The physics of those events and the parameters that apply to the therapeutic use of the isotopes are the primary topics of this report. PMID- 9209530 TI - Influence of brachytherapy applicators geometry on dose distribution in cervical cancer. AB - AIM: Although the relationship between the dose delivered to adjacent organs (urinary bladder and rectum) and the frequency and severity of treatment complications has been reported in many series, the factors influencing pelvic dose distribution are not well defined. The aim of the study was to assess retrospectively the influence of the size of cervical cancer brachytherapy applicators (ovoids and uterine tandems) on pelvic dose distribution and the impact of various therapy-dependent factors on patient anatomy and on dose distribution in particular applications. PATIENTS AND METHOD: The subject of this study were 356 cervical cancer patients treated with Selectron LDR as a part of their radical radiotherapy. Analysed factors included preceding external beam radiotherapy (EBRT) or brachytherapy applications, use of general anaesthesia for application and the system of pellet loading. RESULTS: Significant correlation was found between the size of applicators and doses to bladder, rectum and points B: larger vaginal applicators produced lower dose in bladder and rectum and higher dose in point B (all p < 0.0001), longer uterine tandems produced lower dose in rectum and higher dose in point B (both p < 0.0001). Significant decrease in the frequency of use of large applicators (ovoids: p < 0.0001, tandems: p = 0.055) and worsening of dose distribution, i.e. higher doses to critical organs (respectively: bladder p = 0.0012, rectum p = 0.02) and lower point B dose (p = 0.0001) were observed at consecutive brachytherapy applications. Similar situation occurred in patients, who received EBRT prior to brachytherapy (ovoids: p < 0.001, tandem: p = 0.04, bladder dose: p = 0.009, rectal dose: p = 0.073, point B dose: p = 0.059). Vaginal applicators were larger (p = 0.026) and the dose distribution was better (bladder: p = 0.023, rectum: p = 0.002, point B: p = 0.0001) in patients who had their insertions performed under general anaesthesia. The comparison of 2 consecutively used systems of pellet loading revealed more favourable dose distribution: lower dose for bladder (p = 0.014) and higher dose for point B (p < 0.0001) for the system, which utilised more sources in ovoids and in the distal part of the uterine tandem, in spite of more frequent use of smaller applicators in this group of patients. In multivariate analysis ovoid size was related to preceding external beam radiotherapy (p = 0.025). Uterine tandem length was dependent on the number of preceding intracavitary applications (p < 0.001) and preceding external beam radiotherapy (p = 0.007). Bladder dose was related to preceding brachytherapy (p = 0.011) and the pattern of pellet loading (p = 0.031). Rectal dose was dependent only on the use of general anaesthesia during application (p = 0.001) and point B dose was dependent on the pattern of pellet loading (p < 0.001) and marginally-on the use of preceding external beam radiotherapy (p = 0.06). CONCLUSIONS: The results of this study allow for identification of treatment-related factors determining pelvic dose distribution in cervical cancer brachytherapy and may potentially enable optimisation of this distribution in particular clinical situation. PMID- 9209531 TI - [Current views on Milrinon--a PDE-III-inhibitor]. PMID- 9209532 TI - [Therapy of hypertensive crises. Profile of intravenous Ebrantil]. PMID- 9209534 TI - [Prevention of thromboembolism in ambulatory surgery]. PMID- 9209533 TI - [The role of rhErythropoietin in the framework of methods avoiding donor blood]. PMID- 9209535 TI - [An attenuated corticoid for the treatment of eczema]. PMID- 9209536 TI - [Navoban--the new 5HT3-antagonist]. PMID- 9209537 TI - [Hope for infarct patients. The ACE inhibitor Trandolapril is effective even in high risk patients]. PMID- 9209538 TI - [Comparative praxis study of the effectiveness of heparin in xerophthalmia]. PMID- 9209539 TI - [The AMO-Array multifocal lens. A definite advance in cataract surgery. Report to the PhakoFlex Meeting and the AMO-Array Launch Session, Zermatt Switzerland, 16 17 January 1997]. PMID- 9209540 TI - [Mild and solid. Minimally invasive surgery of the spinal column and stabilization of the lumbar vertebrae in degenerative diseases]. PMID- 9209541 TI - [Vitamin D and calcium in the basic therapy of osteoporosis. It is never too late for therapy]. PMID- 9209542 TI - [Gabapentine--advances in the management of epilepsy]. PMID- 9209543 TI - [ACE inhibitors in cardiac insufficiency and myocardial infarct. What happens after myocardial infarct?]. PMID- 9209544 TI - [Ketoprofen--a first class nonsteroidal anti-arrhythmia agent]. PMID- 9209545 TI - Effects of ethanol on the inner layers of chick retina during development. AB - Optic nerve hypoplasia is an important malformation of the fetal alcohol syndrome whose teratogenic mechanisms are unknown. In our experimental model we have quantified the concentration of ethanol and acetaldehyde in the retina and vitreous humor of the developing chick. The effect of ethanol alone during retinal development was analyzed by conventional histological techniques and by immunostaining. A single injection of ethanol in the vitelline sac at the beginning of retinal cell differentiation retarded synaptogenesis in the inner plexiform layer and produced abundant ganglion cell death and a sharp diminution of myelinic axons. Our observations could help to explain certain alterations described in children exposed to ethanol during the development of their nervous system. PMID- 9209547 TI - Spontaneous preference for ethanol in naive rats is influenced by cholecystokinin A receptor antagonism. AB - Naive adult male Wistar rats free to choose between water or 10% ethanol (v/v) spontaneously became water-preferring (WP) rats, as they drank mainly water (approximately 35 ml per day), or alcohol-drinking (ED) rats, as they also drank a significant amount of ethanol (approximately 14 ml per day). The selective CCKA receptor antagonist L-364,718 at doses selective for the CCKA receptor (5 micrograms/kg, IP) halved the consumption of alcohol of the ED rats without modifying their total liquid in-take. In contrast, the CCKB antagonists L-365,260 or GV150013 were without effect when used at doses selective for the CCKB receptor. These data indicate that the CCK system could be involved in the modulation of alcohol intake. In particular, they suggest that CCKA receptors could play a role in the ethanol preference. PMID- 9209546 TI - Repeated ethanol withdrawal experience increases the severity and duration of subsequent withdrawal seizures in mice. AB - Repeated ethanol withdrawal experience has been shown to result in an exacerbation of future withdrawal episodes. This sensitization of the withdrawal response has been hypothesized to represent a "kindling" phenomenon. The present study was designed to examine whether a systematic increase in the number of previous ethanol withdrawal experiences increases both the severity and duration of a subsequent withdrawal response. An established model of repeated ethanol intoxication/withdrawal was employed in which adult C3H mice were chronically exposed to ethanol vapor in inhalation chambers. In the first experiment, multiple withdrawal (MW) groups of mice received nine (MW x 9), six (MW x 6), or three (MW x 3) cycles of 16-h ethanol vapor separated by 8-h periods of abstinence prior to testing: a single withdrawal (SW) group was tested following a single bout of 16-h ethanol exposure; and a control (C) group did not receive any ethanol treatment throughout the experiment. In a second experiment, a group of mice (MW1-9) were repeatedly tested over nine cycles of withdrawal. A third experiment was designed to assess the effects of repeated pyrazole administration on the potentiated withdrawal seizure response. Results indicated a positive relationship between the number of previously experienced ethanol withdrawals and the severity and duration of a subsequent withdrawal episode. Blood ethanol levels were similar for all ethanol-exposed groups prior to withdrawal assessment. Further, the intensity of withdrawal seizures (handling-induced convulsions) progressively increased over nine cycles of intoxication/withdrawal and repeated testing did not significantly influence the development of this potentiated response. In addition, repeated administration of pyrazole did not appear to influence this withdrawal sensitization phenomenon. Collectively, these results provide further support for the "kindling" hypothesis of ethanol withdrawal. PMID- 9209548 TI - The development of the blood-brain barrier in alcohol-exposed rats. AB - Circulating horseradish peroxidase (HRP) was used as a tracer to determine if the blood-brain barrier to protein was altered by dietary prenatal alcohol exposure. Animals were prepared for light microscopic visualization of HRP after HRP infusion on gestational days 16, 18, 20, 22 and postnatal day 4. There was no consistent evidence of HRP leakage through the BBB in the alcohol-exposed animals compared to control animals. Capillary endothelial cells and perivascular astrocytic endfeet were morphologically characterized by electron microscopy in rat optic nerve and cerebellum following dietary prenatal and postnatal ethanol exposure. Photomontages of optic nerve capillaries from G20 and P5 animals and cerebellar capillaries from P15 animals were examined for evidences of effects of alcohol on the development of the capillaries and adjacent astroglial endfeet. There was no consistent evidence of any alcohol-induced effect that could indicate a disruption of the vessel, the endothelial tight junctions, the perivascular glial limiting membranes, or the extent of vascular ensheathment by astrocytic endfeet. PMID- 9209549 TI - Discriminative stimulus effects of ethanol: lack of antagonism with N-methyl-D aspartate and D-cycloserine. AB - Several drug discrimination studies reported that both competitive and uncompetitive NMDA receptor antagonists substituted for ethanol stimulus in rats. In the present study we examined if compounds that act as agonists at the NMDA receptor complex, D-cycloserine (a partial agonist at the glycine positive modulatory site) and N-methyl-D-aspartate (an agonist at the glutamate binding site), could antagonize the discriminative stimulus effects of ethanol. Rats were trained to discriminate between IP administered 1.0 g/kg of ethanol (10% v/v) and saline under a sweetened milk-reinforced fixed ratio 10 (FR10) schedule of reinforcement. When the animals met the discriminative criteria, antagonism tests were conducted with D-cycloserine (0.3-10.0 mg/kg, IP) and N-methyl-D-aspartate (15.0-60.0 mg/kg, IP). Neither D-cycloserine nor N-methyl-D-aspartate antagonized the ethanol-mediated discriminative stimulus effects. In addition, D-cycloserine (3.0-300.0 mg/kg, IP) did not substitute for ethanol. These results indicate that at least certain agonists at the NMDA receptor complex do not attenuate the ethanol interoceptive cue in the rat. PMID- 9209550 TI - Intracellular mechanisms of constriction of rat aorta by ethanol. AB - The intracellular mechanisms mediating vasoconstriction by ethanol are poorly understood. This investigation was designed to provide evidence on the role of protein kinase C (PKC) and calmodulin in vasoconstriction by ethanol. We studied helically cut strips of rat aorta that were exposed to ethanol before and in the presence of the PKC inhibitors calphostin C (79, 239, and 798 nM) or 1-(5 isoquinolinylsulfonyl)-2-methylpiperazine (H7, 10 microM), and the calmodulin inhibitor, trifluoperazine (TFP, 10 microM). To test for the specificity of the PKC inhibitors, we measured the responses of aortas to potassium and phorbol 12 myristate 13-acetate (PMA) in the absence and presence of calphostin C and H7. To test for the specificity of TFP, we measured the responses of aortas to serotonin, potassium, PMA, and the thromboxane A2 mimic. 9,11-dideoxy-11 alpha, 9 alpha-epoxy-methanoprostaglandin F2a (U46619), in the absence and presence of TFP. We also studied the effect of the combination of calphostin C and TFP on constriction of the aorta by ethanol. We also measured the importance of intracellular and extracellular calcium in constriction of the aorta by ethanol. Force generation was measured before, and then during exposure of the strips to calcium-free buffer with EGTA, or calcium-free buffer with EGTA plus caffeine. We found that both PKC inhibitors antagonized vasoconstriction by ethanol and PMA. However, H7 antagonized contractions by potassium, but calphostin C did not. We found that TFP caused 99 +/- 1% inhibition of maximum contraction to serotonin, 90 +/- 4% inhibition of maximum contraction to potassium, 63 +/- 6% inhibition of maximum contraction to PMA, and 8 +/- 5% inhibition of maximum contraction to U46619. TFP caused a 22 +/- 8% inhibition of contraction to ethanol. The combination of TFP and calphostin C antagonized vasoconstriction by ethanol to a degree similar to that of calphostin C alone. We also found that contractions to ethanol were only 16 +/- 7% of control values in a calcium-free plus EGTA buffer. Contractions to ethanol were 0 +/- 1% of control values in calcium-free buffer with EGTA plus caffeine. We conclude that: 1-vasoconstriction by ethanol is, at least in part, mediated by PKC; 2-constriction by ethanol is mediated to a minimal extent by calmodulin, and 3-part of the constriction by ethanol of the aorta is mediated by a caffeine-sensitive pool of intracellular calcium. PMID- 9209551 TI - The effect of ethanol drinking on opioid analgesia and receptors in mice. AB - Recent studies suggest substantial interactions between opioids and ethanol (EtOH). Both in vivo and in vitro experiments indicate that EtOH can regulate opioid systems and that opioids can modify EtOH consumption. In the present studies, we examined if EtOH consumption altered opioid receptors and the potency of opioid analgesics. Mice were given unlimited access to 6-7% EtOH alone for 7 days or were allowed to drink increasing concentrations (3-6%) of EtOH over 13-14 days. Controls had access to water. The EtOH groups drank significantly less volume than controls, although there were no significant differences in body weight or baseline nociception. The analgesic (tail flick) potency of SC morphine was decreased by approximately 1.6-2.0-fold in EtOH-treated mice. A single acute dose of EtOH (1 g/kg) that produced blood alcohol levels in excess of that for 7 day exposure to EtOH, did not change morphine's analgesic ED50, suggesting that chronic exposure to EtOH was necessary for the reduction in potency. The change in morphine potency was not due to pharmacokinetic differences because EtOH consumption did not modify the concentration of morphine in brain and spinal cord. The analgesic potency of a delta-opioid receptor agonist (ICV DSLET) was also decreased by approximately 2-fold. Saturation binding studies indicated no changes in the density or affinity of brain and spinal cord delta-opioid ([3H]DPDPE, [3H]DSLET, [3H]DeltorphinII) and mu-opioid ([3H]DAMGO) receptors. Similarly, there was no significant effect of EtOH on delta-opioid receptor mRNA in either brain or spinal cord preparations. Taken together, these data suggest that EtOH consumption decreases the analgesic potency of opioids in mice through a mechanism that is unrelated to pharmacokinetics or opioid receptor changes in brain and cord. PMID- 9209552 TI - Chronic treatment of cultured cerebral vascular smooth cells with low concentration of ethanol elevates intracellular calcium and potentiates prostanoid-induced rises in [Ca2+]i: relation to etiology of alcohol-induced stroke. AB - The influence of chronic treatment of cultured canine cerebral vascular smooth muscle cells, with low concentrations of ethanol, on the intracellular concentrations of free calcium ([Ca2+]i) was studied by use of the fluorescent indicator, fura-2, and digital imaging microscopy. The resting level of [Ca2+]i in the cerebral vascular smooth muscle cells was 89 +/- 3.2 nM. Exposure of these cells to 10 and 25 mM ethanol for 5 days resulted in significant elevation of [Ca2+]i (mean rises to 208 +/- 11.4 and 307 +/- 14.0 nM, respectively), and potentiated the transient rise in [Ca2+]i induced by 10(-7) M PGF2 alpha. However, exposure of these cerebral cells to a high-concentration ethanol (100 mM) resulted in only a slight increase of [Ca2+]i (106 +/- 6.9 nM) and lack of effects on the [Ca2+]i response to PGF2 alpha. Irrespective of the different ethanol treatments, the subcellular distribution of [Ca2+]i was heterogeneous in all the cells tested. Our data suggest that chronic exposure of cerebral vascular smooth muscle cells to ethanol, particularly at low concentrations, results in dramatic increases in [Ca2+]i and the responses of these vascular smooth muscle cells to prostanoids. These results support an hypothesis whereby ethanol induces stroke by causing spasm and rupture of cerebral blood vessels as a consequence of large rises in intracellular Ca2+. PMID- 9209553 TI - Acute alcohol tolerance in social drinkers: changes in subjective effects dependent on the alcohol dose and prior alcohol experience. AB - Using subjective ratings of the degree of alcohol intoxication, the interaction between the drinking history of the subjects, the alcohol dose, and acute alcohol tolerance were examined in light and moderate alcohol consumers (N = 10). Both groups of subjects were tested with doses of alcohol corresponding to 0.5 and 1.0 g/kg. Dose order was random and tests were carried out with an interval of 1 week. Reports of the subjects' previous experience with these doses of alcohol indicated that the moderate consumers ingested the lower (but not the higher) of the doses quite regularly, whereas light consumers were rather inexperienced with both of the doses. Comparison of blood alcohol concentrations as measured by breath and blood analysis yielded slightly different results, the concentrations being significantly higher as measured by breath analysis. This result was mainly associated with the initial phases, where this difference was greatest. Acute tolerance was assessed by comparing the ratings at equal concentrations of alcohol on the ascending and the descending limbs of the alcohol concentration curve. Due to the lag in the measurements of breath and blood alcohol concentrations, the outcome of the evaluations of acute tolerance was also influenced by whether breath or blood alcohol concentrations were used to obtain similar concentrations in both phases. Results based on the breath alcohol concentrations showed that in light alcohol consumers, acute tolerance was demonstrated for both of the doses. In moderate alcohol consumers only the higher of the doses produced evidence for acute tolerance. However, if comparisons are based on blood alcohol concentrations, moderate alcohol consumers also show an apparent acute tolerance for the lower of the doses tested. The present results clearly demonstrate the complexity of the acute tolerance phenomenon, and emphasize the fact that the results are dependent on the dose of alcohol, the subjects' prior experience with alcohol as well as the procedure used for measuring alcohol concentration. PMID- 9209554 TI - Effects of sucrose-substitution initiation on patterns of drinking by Lewis rats during continuous alcohol access. AB - Initiation of alcohol drinking using the sucrose-substitution procedure was studied in inbred Lewis rats. One group of animals was initiated to self administer alcohol prior to being placed in the continuous-access condition, whereas the second group of animals did not undergo initiation. During the continuous-access period, the animals were housed in operant chambers where they had continuous access to alcohol (10% v/v), food, and water during daily 23-h experimental sessions. After 5 weeks of baseline conditions, the response, requirement for food was increased over weeks. This was followed by weekly increases in the ethanol response requirement with the food response requirement returned to baseline conditions. In the continuous-access condition, both groups consumed similar amounts of alcohol by the end of the 4-week baseline period and showed similar numbers of dippers presented per alcohol bout and number of alcohol bouts per day. During the food response requirement manipulation, alcohol consumption increased for both groups but intake increased significantly more for the noninitiated group. The difference between groups was accounted for by a larger number of alcohol drinking bouts per day for the noninitiated group. Alcohol consumption decreased at each increase in ethanol reinforcement response requirement for both groups. Alcohol-reinforced responding per session increased for the noninitiated animals but remained unchanged for the initiated group during this condition. Responding increased substantially for both groups when the alcohol reinforcement response requirement was returned to baseline conditions. These results suggest that alcohol may serve more as a food source for noninitiated animals during the food reinforcement manipulation and that initiation may result in more resistance to change during the alcohol reinforcement manipulation. These data show that the type of initial exposure to alcohol can impact future drinking patterns. PMID- 9209555 TI - Ethanol teratogenesis in the C57BL/6J, DBA/2J, and A/J inbred mouse strains. AB - Research has shown variations in susceptibility to alcohol-related birth defects in humans. Genetic differences are one reason for this variability. This study compared three inbred mouse strains to determine whether they differ in their susceptibilities to ethanol teratogenesis because previous studies have generated conflicting data. Pregnant C57BL/6J (B6), DBA/2J (D2), and A/J (A) dams were intubated intragastrically with either an acute dose of ethanol (5.8 g/kg) or an isocaloric amount of maltose-dextrine on day 9 of pregnancy. Litters were removed on day 18 of pregnancy and examined for gross, soft-tissue, and skeletal malformations. Results showed that ethanol-exposed B6 litters had a higher percentage of digit (19%), kidney (24%), and skeletal (32%, mostly vertebral) malformations than their maltose-exposed controls (7% or below). Prenatal exposure to ethanol increased skeletal (68%, both rib and vertebral) malformations for A litters when compared to their maltose-exposed controls (4%), but did not increase digit or kidney malformations. Ethanol-exposed D2 litters did not differ from maltose-exposed controls. Maternal blood ethanol levels did not differ among the B6, D2, and A strains. These results provide additional evidence suggesting a genetic component to ethanol teratogenesis. PMID- 9209557 TI - The effects of ethanol on a measure of conditioned fear in mice. AB - The effects of ethanol on conditioned freezing, a species-specific defensive behavior used as an assay of fear, was examined in mice. In Experiment 1, ethanol, 1.2 g/kg, significantly increased freezing compared to a saline control when the mice were reexposed to a context in which they were previously shocked. Experiment 2, which administered ethanol or saline to no-shock control animals, demonstrated that the potentiated freezing produced by ethanol in Experiment 1 was specific to the interaction of ethanol and the stress response. These results suggest that both the qualitative and quantitative dimensions of the environmental stressor, as well as the dose of ethanol used, may be critical for determining ethanol's effect on a stress response. PMID- 9209556 TI - Acute treatment of paternal alcohol exposure produces malformations in offspring. AB - Male rats were intubated only once with either 6, 4, 2, or 0 g/kg alcohol. Food was removed from all animals for several hours after intubation. Males were bred with a single female until sperm was observed in the vaginal smear, for up to a maximum of 7 days after treatment. Females were sacrificed on gestation day 20. There were no significant effects on mating, fecundity, or litter size, but there were significant dose-response increases in "runts" and significant linear associations in the number of malformations in alcohol-sired offspring. A second study using the same methodology found similar results. The results indicate that a single acute treatment with alcohol just prior to breeding may have a significant effect on offspring. PMID- 9209558 TI - Toward a definition of a valid model of alcoholism: multiple animal models for multiple diseases. PMID- 9209559 TI - Progress in Alzheimer's disease. Pause and perspective. PMID- 9209560 TI - The subjective burden of depression. PMID- 9209561 TI - Index of affective suffering. Linking a classification of depressed mood to impairment in quality of life. AB - The authors report on development and validation of their Index of Affective Suffering (IAS), an explicit measure of subjective symptom severity, constructed as a typological system, rather than a multifactorial scoring system, with combinations of "intensity" (degrees of distress per given time) and "extensity" (duration and frequency of episodes, and number and variety of life events and activities that are pervaded by distress) item rankings. Seven levels of severity appeared to have both face value and empirical justification. Further conceptual and methodological advances would enhance the development and evaluation of treatments for geriatric mental disorders and chronic diseases in general. PMID- 9209562 TI - Clinical features of obsessive-compulsive disorder in elderly patients. AB - There has been no systematic study of the clinical features of obsessive compulsive disorder (OCD) in elderly patients. This study describes the symptoms and characteristics of OCD among 32 outpatients age 60 or older and 601 younger patients meeting DSM-III-R criteria and given the Yale-Brown Obsessive-Compulsive Scale (YBOCS), NIMH scale, and a 41-item symptom questionnaire. Elderly patients had a later age at onset compared with younger patients. No differences were found in severity of symptoms on the YBOCS. Elderly patients had fewer concerns about symmetry, need to know, and counting rituals. Handwashing and fear of having sinned were more common. There were few differences in clinical features of OCD among the elderly patients compared with younger OCD patients. PMID- 9209563 TI - Screening for depression in older persons with low vision. Somatic eye symptoms and the Geriatric Depression Scale. AB - The authors compared the sensitivity and specificity of a somatic eye symptoms question with the Geriatric Depression Scale (GDS) to detect depression in older patients with low vision. The sample was 70 patients (65+ years) attending a low vision clinic. A geriatric nurse-practitioner examined all patients, diagnosed major depression by DSM-III-R criteria, and completed the GDS and an eye symptoms (ES) questionnaire. Twenty-seven patients (38.6%) met criteria for major depression. Although the sensitivity of the ES and GDS were identical (63%), the GDS was more specific (77% vs. 54%) and had higher positive predictive value (63% vs. 46%). Both measures identified patients who were more functionally disabled than patients without ES or depression, but with similar vision. Depression frequently accompanies visual impairment in this population. Inquiring about ES appears to be less specific than the GDS, although neither method is highly sensitive. However these screening tools identify distressed persons whose disability is high relative to the severity of their vision loss. PMID- 9209564 TI - A descriptive study of elderly community-dwelling alcoholic patients in the rural south. AB - Alcohol-related disorders, estimated to be more prevalent in the South, are associated with serious comorbid disorders, such as depression and suicide. In a rural outreach program, the authors examined patients with a diagnosis of alcoholism and compared them with nonalcoholic patients on various demographic and descriptive variables. Of 166 patients referred to the program, 35 (21.1%) had an alcohol-related disorder. Alcoholism was significantly associated with male gender and younger age, but nearly half of the alcoholic subjects were women. Alcoholism is associated with inappropriate health care utilization; alcohol-related disorders produced significantly more emergency room visits and somewhat more hospital admission; these patients were less likely to have a primary care physician. No patient was receiving treatment for alcoholism. PMID- 9209565 TI - Generalized anxiety and depression. Assessment over 2 years after stroke. AB - The authors examined the course of anxiety up to 2 years after stroke in relation to depressive symptoms, impairment in activities of daily living (ADLs), and social functioning. One hundred forty-two patients were evaluated at 3, 6, 12, and 24 months after stroke. Anxiety was associated with greater depression severity at all follow-up visits. Depression severity was associated with impairment in ADLs at followup; association of anxiety and impairment in ADLs was present only at the intake visit, with independent effects only for women. Women reported more symptoms of both anxiety and depression during the 2-year period. Younger patients reported more anxiety symptoms, but there was no difference between age-groups in depressive symptoms. Severity of anxiety was also related to higher depression scores at initial hospitalization, but not in the remainder of the 2-year period. In summary, anxiety is associated with increased severity of depressive symptoms and greater impairment in function primarily during the acute hospitalization period. Women and younger patients also may be more vulnerable to anxiety after stroke. PMID- 9209566 TI - The first Summer Research Institute in Geriatric Psychiatry. AB - In July 1995, the American Association for Geriatric Psychiatry (AAGP) sponsored the first annual week-long Summer Research Institute (SRI) in Geriatric Psychiatry, at the University of California, San Diego. The NIMH-funded SRI was intended for promising postresidency and postdoctoral fellows, as well as junior faculty persons interested in research careers in geriatric psychiatry. The SRI focused on the tools needed to begin, maintain, and succeed on that career path and has been followed by continued communication between trainees and faculty. The SRI was highly successful, judging from the participants' evaluations, as well as the trainees' accomplishments in terms of publications and research funding during 1 year of follow-up. The SRI provides a useful model for an approach to bridging and shortening the transition period from fellowship to first research funding and of ensuring a continued flow of new investigators in geriatric psychiatry. PMID- 9209567 TI - Control-relevant intervention in the treatment of minor and major depression in a long-term care facility. AB - The authors assessed the effect of a control-relevant psychosocial intervention in 31 nursing home residents with either major depressive episode or minor depression. An initial group of 22 residents were randomized to either active treatment or waiting list. Four of 11 residents randomized to active treatment were deemed Responders, compared with 0 of 11 on the waiting list (P < 0.05). Of the total of 31 residents who participated in the intervention, 14 (45%) were deemed Responders during the intervention period. For these Responders, the Hamilton Rating Scale for Depression (Ham-D) and Geriatric Depression Scale scores improved significantly during the intervention. The improvement in the Ham D was not sustained 2 months after intervention was terminated. These findings suggest that a psychosocial intervention enhancing socialization according to each resident's choice had a positive therapeutic impact on almost half of the nursing home residents with major or minor depression. However this effect could not be sustained by the residents without the support of the structured program. PMID- 9209568 TI - Can the sensitivity of the Alzheimer's Disease Assessment Scale be increased? AB - The author analyzed and compared Mini-Mental State Exam and Alzheimer's Disease Assessment Scale (ADAS) scores for 48 patients with Alzheimer's disease to determine whether improvements could be made in the ADAS. Results suggest that cognitive ability could be more accurately evaluated by shortening the number of words on the ADAS word recall task and reducing the number of trials on the word recognition task. These modifications would increase the likelihood of identifying medication effects. PMID- 9209569 TI - Comorbidity of dementia and psychiatric disorders in older persons. AB - To further investigate the relationship between psychiatric disorders and dementia in elderly patients, the authors drew a population-based, age-stratified random sample from residents of Rochester, Minnesota, age 65 and older. A trained paramedic completed a 90-minute screening interview, including the Symptom Checklist-90, Mini-Mental State Exam, and Auditory-Verbal Learning Test. Persons failing the screens were interviewed by a psychiatrist and a neurologist. DSM-III R diagnoses were assigned for dementia and other psychiatric disorders. Of 201 participants, 37 were evaluated further by both neurologist and psychiatrist. One received a psychiatric diagnosis alone. Dementia alone was present in four people. Concurrent psychiatric diagnoses and dementia were found in 17 subjects. Much of the psychopathology found in older persons occurs in people with cognitive impairment. Current diagnostic nosology may not be able to capture the interrelatedness of psychiatric syndromes and cognitive impairment in elderly patients. PMID- 9209570 TI - Syndrome of inappropriate antidiuretic hormone (SIADH) in an 80-year-old woman given clomipramine. AB - The author describes the syndrome of inappropriate antidiuretic hormone, with serum sodium decreasing from 137 mEq/L to 122 mEq/L in an 80-year-old woman prescribed clomipramine. The patient's delirium and serum sodium were slow to normalize, prolonging her hospital stay and delaying treatment of her depression. PMID- 9209571 TI - Patient/residents in the community. PMID- 9209572 TI - Genetic variability in two Brazilian ethnic groups: a comparison of mitochondrial and protein data. AB - Sequence data from the first hypervariable segment of the mitochondrial DNA control region of 124 subjects belonging to three African-Brazilian and three Brazilian Indian populations were compared with information related to 12 protein genetic loci from 601 persons living in the same localities. There is high diversity among the mtDNA sites, and the most variable in one ethnic group are not the most variable in the other. No differences in gene diversity between populations within ethnic groups were observed, but the Indians showed a reduced variability. Much more interpopulation variation was observed in the mtDNA data than in the protein set. The relationships obtained for the six populations, however, are the same regardless whether mtDNA or protein loci are considered. African-Brazilians from Porto Alegre and Salvador, situated 3,000 km apart, are more similar to each other than both are to Paredao, despite the geographical proximity between Porto Alegre and Paredao, which are just 50 km apart. The tree topology in relation to the three Indians groups, on the other hand, is that expected when languages, culture, and geography are considered. PMID- 9209573 TI - Whole body bone, fat and lean mass in children: comparison of three ethnic groups. AB - We measured whole body bone, fat and lean mass, by dual-energy x-ray absorptiometry, of third-grade children in a suburban public school district adjacent to Detroit. Of 1,340 eligible children, 773 participated. Using U.S. Census categories, parents identified their children as black/African-American (57%), white (38%), or one of several other categories (5%). Some of the participants also identified with a relatively large Middle Eastern subgroup (Chaldeans). Of the 773 participants, 734 are included in this report (71 Chaldeans, 226 whites, and 437 black/African-Americans; other categories are omitted). We describe body size, body composition, and physical activity levels in the three groups. The Chaldean and black children have significantly higher average whole body bone mineral content (BMC) than whites (P > 0.05), but are not different from each other. Lean mass and height are significantly greater for Chaldeans and blacks than for whites. The ratio of BMC to height was also significantly greater in Chaldeans and blacks compared with whites. Chaldeans have a significantly higher weight and fat mass than either the black or white children, and report significantly less physical activity than either the white or the black children. The higher bone mass among the Chaldean children may be partially explained by their greater body mass, but there is no readily apparent explanation for the observed ethnic differences in body size. We cannot exclude genetic or environmental factors not evaluated in this observational study. Our unexpected finding that Chaldean children, when analyzed as a separate group, are more similar in body composition to black/African-American than to white children contributes to a growing body of literature indicating that the uncritical use of "race" categories may obscure rather than facilitate the identification of population differences. PMID- 9209574 TI - Affiliative relationships between adult males and immature group members in naturally occurring ringtailed lemurs (Lemur catta). AB - Affiliative relations between adult males and immature group members were studied in three naturally occurring groups of ringtailed lemurs (Lemur catta) over a 12 month period at Beza-Mahafaly Reserve, southwestern Madagascar. No statistically significant difference was found in rates of affiliative interactions between adult males and older immatures (juveniles and adolescents) over reproductive seasons. However, some of the adult males in two focal groups increased their rates of affiliative behaviors with older immatures during the lactation and subsequent 1993 postmigration periods. Female involvement with infants during lactation season, the increasing independence of juveniles and adolescents, and length of male tenure may be factors contributing to such a pattern. Neither adult males nor immatures were significantly responsible for the initiation and maintenance of proximity in male-immature dyadic interactions. Neither dominance rank nor age-class of the adult male affected their rates of affiliative behavior with immatures. The majority of focal males exhibited an interest in infants and occasionally participated in alloparental care. It is suggested that adult males can benefit from affiliative relations with immatures in terms of opportunities for greater spatial centrality in the group, which can lead to enhanced predator protection and greater opportunities to develop affiliative relationships with females. Immatures can benefit from affiliation with males in relation to enhanced predator detection and protection, alloparental care, and opportunities to develop social skills. PMID- 9209575 TI - Volumetric comparisons in the cerebellar complex of anthropoids, with special reference to locomotor types. AB - Seven measurements in the cerebellar complex were completed on 45 individuals, including 26 species of anthropoids from Stephan's collection. These included 12 species of New World monkeys, 10 species of Old World monkeys, and Hylobates, Gorilla, Pan, and humans. The measurements were the volume of medial (fastigial) (CM), interpositus (globose and emboliform) (CI), and lateral (dentate) (CL) cerebellar nuclei, ventral pons (VPo), inferior olivary principal (OLIPr), and accessory (OLIAc) nuclei and vestibular nuclear complex (VES). The relative size of each nucleus was expressed in size indices based upon the allometric line obtained by the reduced major axis analysis. The indices of three cerebellar nuclei reflect the relative size of three longitudinal zones of the cerebellum. The cerebellar hemisphere-lateralis zone is represented by the CL indices, the vermis-medialis zone by the CM indices, and the pars intermedius-interpositus zone by the CI indices. The results show that the VPo and OLIPr indices are closely related to the CL indices. This lateral zone group of nuclei is the most progressively developed in humans, whereas the CM, CI, OLIAc, and VES are independent of the developmental trend manifest by the lateral zone group of nuclei. The indices are discussed in relation to the predominant locomotor pattern exhibited by a species. The size indices of arboreal quadrupeds show a development of all nuclei in the cerebellar complex. This is interpreted as indicating that arboreal monkeys live in complicated, discontinuous, three dimensional space and need exceptional cerebellar capacity for each pattern of locomotion and positional behavior. Progressive development of the lateral zone group of nuclei only compared to other nuclei was recognizable in humans. This development is considered to be related not to bipedalism, but to versatile and coordinated finger movement, resulting after bipedalism was established. This cerebellar reorganization is also a prerequisite (Leiner et al. [1993] TINS 16: 444-447) for the evolution of human language. The differences between size indices of the nuclei of Macaca (= pronograde primate) and Ateles (= antipronograde one) are compared in relation to their vertical climbing kinesiological data. PMID- 9209576 TI - Osteopenia and stable isotope ratios in bone collagen of Nubian female mummies. AB - Stable carbon and nitrogen isotopes were analysed on bone collagen of 43 Sudanese Nubians from the X-Group period to test dietary hypotheses for the high frequency of osteopenia in this population. Stable carbon isotope ratios indicate that both normal and osteopenic individuals consumed the same mixed diet of C3 and C4 sources, which are assumed to have been constituted by the grain staples wheat/barley and sorghum/millet respectively. Females with osteopenia, however, have significantly elevated delta 15N values. The enrichment effect is greatest in the third and fifth decades of life, and is consistently patterned with microstructural and frequency differences previously reported by other researchers. It is suggested that delta 15N is reflecting differences in urea excretion and the renal processing and clearance of calcium and phosphorus. The study not only alerts us to the susceptibility of stable nitrogen isotopes to non dietary (i.e. physiological) factors, but also identifies nitrogen isotope ratios as a possible new marker for osteopenia. PMID- 9209577 TI - Cribra orbitalia and trace element content in human teeth from Neolithic and Early Bronze Age graves in southern Poland. AB - Determination of element levels in bones and teeth can complement knowledge of the diagnostics and etiology of various diseases in prehistoric populations. Calcium (Ca), cadmium (Cd), copper (Cu), iron (Fe), and lead (Pb) content were analyzed in teeth from human skeletons dated to 3,000-1,400 BC from Malopolska Upland loess. Levels of iron and calcium were determined using atomic absorption spectroscopy (AAS), and lead, cadmium, and copper levels were measured using anodic stripping voltametry (ASV). Molar teeth from specimens with cribra orbitalia were selected for analyses, and teeth from specimens with no pathological changes were used as a control. No significant correlations between the content of particular elements and the tooth class, specimen age, or depth of burial pit were observed. The Fe content in specimens with cribra orbitalia is not the best measure for this disease's etiology. Thus, interelement correlations and proportions might give a better picture of the biological condition of the specimen and of the investigated groups. PMID- 9209578 TI - Harris lines: a study of age-associated bias in counting and interpretation. AB - Harris lines are regularly used in paleopathology as indicators of episodic nonspecific stress, but the methodology for their use has not been clearly established. We studied radiographs of the distal shaft of the tibia in 82 immature and 49 mature subjects from a medieval burial site and compared the number of Harris lines and observer error according to age categories. We found statistically significant differences in both line counts and in observer error by age groups. In conclusion, studies of Harris lines must take into account age variation in order to be validated. PMID- 9209579 TI - Neandertal capitate-metacarpal articular morphology. AB - Neandertal capitate-metacarpal 2 and 3 articulations have been observed to differ in orientation and shape from those of more recent humans. To evaluate this, we tested for differences in capitate-metacarpal 2 (MC2) and MC2-capitate facet orientations and MC2 and MC3 robusticity indices, and for multivariate shape equivalence of the capitate-MC2/MC3 facets and the MC3 diaphysis and styloid process between samples of Neandertals and recent humans. Canonical discriminant functions of log size- and-shape and log shape transformed measurements were run on variables of the capitate-MC2 and MC3 facets, and these plus MC3 diaphysis and styloid process variables. The null hypothesis of shape equivalence is rejected for both variable sets. Modern human capitate-MC morphology results from nonallometric increases in distal capitate breadth and the projection of the MC3 styloid process, and reductions in MC2 facet height and MC3 facet breadth. These shape changes are associated with a significantly less parasagittal orientation of the capitate-MC2 facets in recent humans, but are only trivially correlated with MC 2 and 3 robusticity indices. The recent human capitate-MC 2 and 3 morphology may reflect a shift in habitual joint reaction forces from more axial to more oblique forces while maintaining similar pronation/supination of the MC2. However, the full behavioral implications of these contrasts remain unclear. PMID- 9209580 TI - Systematic assessment of a maxilla of Homo from Hadar, Ethiopia. AB - The Hadar site in Ethiopia is a prolific source of hominid fossils attributed to the species Australopithecus afarensis, which spans the period 3.4-3.0 million years (myr) in the Sidi Hakoma, Denen Dora and lower Kada Hadar Members of the Hadar Formation. Since 1992 a major focus of field work conducted at Hadar has centered on sediments younger than 3.0 myr, comprising the bulk of the Kada Hadar Member. Witnessing the rise of the "robust" Australopithecus clade(s), the origin of Homo, and the first record of lithic artifacts, the period between 3.0 and 2.0 myr is strategically vital for paleoanthropology. However, in eastern Africa it is a particularly poorly sampled temporal interval. This paper provides a detailed comparative description of a hominid maxilla with partial dentition found at Hadar in 1994. The specimen, A.L. 666-1, derives from a lithic artifact bearing horizon high in the Kada Hadar Member, 0.8 m below the BKT-3 tephra, dated by the 40Ar/39Ar method to 2.33 +/- 0.07 myr. Our preliminary investigation of the hominid specimen showed unambiguous affinities with early representatives of the Homo clade (Kimbel et al. [1996] J. Hum. Evol. 31:549-561). Further studies on maxillary and dental morphology lead us to attribute A.L. 666-1 to Homo aff. H. habilis. The new Hadar jaw is the first paleontological evidence for the projection of the H. habilis maxillofacial morphotype well back into the Pliocene. It may represent a male of this species, whose maxillary hypodigm consists chiefly of females. A subsidiary finding of our study is that of the three earliest recorded species of Homo (H. habilis, H. rudolfensis, H. erectus), it is H. habilis that exhibits facial morphology closest to that expected in their last common ancestor. PMID- 9209581 TI - The expanded mandibular condyle of the Megaladapidae. AB - The Megaladapidae have a posterior expansion of the articular surface of the mandibular condyle. Several other strepsirhine species exhibit a similar condylar surface. In this study, I propose two behavioral scenarios in which the posterior articular expansion might function: 1) contact with the postglenoid process and resistance to joint stress during browsing, and 2) movement against the postglenoid process during the fast closing and power strokes of mastication, as a consequence of large transverse jaw movements and associated with a strong mandibular symphysis. These models are evaluated through dissection of the TMJ in Lepilemur and from comparative anatomical observations on strepsirhines and ungulates. In Lepilemur the mandibular symphysis is unfused, but compared to the unfused symphyses of other strepsirhines is strengthened by interlocking bony projections (Beecher [1977] Am. J. Phys. Anthropol. 47:325-336). An accessory articular meniscus is found between the posterior articular expansion and the postglenoid process in Lepilemur, suggesting that significant movement occurs in this part of the TMJ. The symphysis is fused in adult specimens of Megaladapis. A posterior articular expansion is common among ungulates, and its presence is associated not with browsing but with symphyseal fusion. This supports the second model and suggests that the posterior articular expansion functions as a movement surface during mastication. Schwartz and Tattersall ([1987] J. Hum. Evol. 16:23 40) cite the posterior articular expansion as a synapomorphy uniting an Adapis Leptadapis clade with a Megaladapidae-Daubentonia-Indridae clade. The comparative evidence suggests that the posterior articular expansion has evolved convergently in adapines, notharctines, megaladapids, hapalemurids, and indrids as part of a functional complex related to herbivory. However, close morphological similarity of the posterior articular expansion among genera within these strepsirhine subfamilies and families indicates that it is probably a reliable synapomorphy at lower taxonomic levels. PMID- 9209582 TI - A new reconstruction of RUD 77, a partial cranium of Dryopithecus brancoi from Rudabanya, Hungary. AB - A newly reconstructed cranium (RUD 77) of the Miocene fossil hominoid Dryopithecus, formerly Rudapithecus (Kretzoi [1969] Symp. Biol. Hung. 9:3-11; Begun and Kordos [1993] J. Hum. Evol. 25:271-286) is presented here. This specimen, from the late Miocene locality of Rudabanya, in northeastern Hungary, consists of portions of the neurocranium, face, and postcanine dentition. Newly recovered portions of the parietal, occipital, temporal, zygomatic, and premaxillary bones, which are described here for the first time, in association with previously described portions of this specimen (Kordos [1987] Ann. Hist. Nat. Mus. Natl. Hung. 79:77-88) make RUD 77 among the most complete and well preserved neurocrania of any Miocene hominoid. Detailed anatomical descriptions and measurements are provided here, along with comparisons to other relatively complete Miocene hominoid cranial remains, and to living hominoids. While a more complete phylogenetic analysis is in preparation based on the sample as a whole, it is suggested here that RUD 77 provides some additional evidence in support of a previous hypothesis that Dryopithecus is more closely related to the African apes and humans than is Sivapithecus. PMID- 9209583 TI - Radiation and speciation of spider monkeys, genus Ateles, from the cytogenetic viewpoint. AB - The chromosomes of 22 animals of four subspecies of the genes Ateles (A. paniscus paniscus, A. p. chamek, A. belzebuth hybridus, and A. b. marginatus) were compared using G/C banding and NOR (nucleolar organizer region) staining methods. The cytogenetic data of Ateles in the literature were also used to clarify the phylogenetic relationships of the species and subspecies and to infer the routes of radiation and speciation of these taxa. Chromosomes 6 and 7 that showed more informative geographic variation and the apomorphic form 4/12, exclusively in A. p. paniscus, are the keys for understanding the evolution, radiation, and specification of the Ateles taxa. The ancestral populations of the genus originated in the southwestern Amazon Basin (the occurrence area of A. paniscus chamek) and spread in the Amazon Basin and westward, crossing the Andes and colonizing Central America and northwesternmost regions of South America. The evolutionary history of the northern South American taxa is interpreted using the model of biogeographical evolution postulated by Haffer [Science 185:131-137, 1969]. Ateles paniscus paniscus is the genetically most differentiated form and probably derives from A. belzebuth hybridus. Based on the karyotype differences, the populations of Ateles can be divided into four different groups. These findings indicate the necessity of a more coherent taxonomic arrangement for the taxa of Ateles. PMID- 9209584 TI - Determinants of female dispersal in Thomas langurs. AB - Female dispersal occurs in a number of primate species. It may be related to: avoidance of inbreeding, reduction in food competition, reduction of predation risk, or avoidance of infanticide in combination with mate choice. Female dispersal was studied for a 5-year period in a wild population of Thomas langurs (Presbytis thomasi) that lived in one-male multi-female groups. Juvenile and adult individuals of both sexes were seen to disperse. Females appeared to transfer unhindered between groups, mostly from a larger group to a recently formed smaller one. They transferred without their infants and when not pregnant, and seemed to transfer preferentially during periods when extra-group males were harassing their group. During these inter-group encounters extra-group males seemed to try to commit infanticide. Thus, the timing of female transfer was probably closely linked to infanticide avoidance. Moreover, females seemed to transfer when the resident male of their group was no longer a good protector. The observations in the present study suggest that females transferred to reduce the risk of infanticide. Female dispersal may have another ultimate advantage as well, namely inbreeding avoidance. Due to the dispersal of both females and males the social organization of Thomas langurs was rather fluid. New groups were formed when females joined a male; male takeovers were not observed. Bisexual groups had only a limited life span, because all adult females of a bisexual group could emigrate. This pattern of unhindered female dispersal affects male reproductive strategies, and in particular it might lead to infanticidal behavior during inter-group encounters. PMID- 9209585 TI - Simple sequence repeat (SSR) polymorphisms for colony management and population genetics in rhesus macaques (Macaca mulatta). AB - Cross-species amplification of 72 SSR (predominantly tetranucleotide) loci from the DNA of six rhesus macaques of diverse regional origins was conducted using human primers for the polymerase chain reaction (PCR). Thirteen of these primer pairs, which consistently and unambiguously amplified polymorphic fragments from these six samples and which exhibited Mendelian properties, were also used to amplify SSR loci for 176 male rhesus macaques that are founders of six different captive breeding colonies. These include four groups of macaques originating in India and one group of macaques each that originated in China and Thailand. Gene diversity based on the SSR loci provided a reliable estimate of average heterozygosity but was between two and four times higher than that for 9 protein coding loci. Based on the SSR loci, Chinese rhesus were more genetically diverse and unique than were rhesus from India or Thailand, a conclusion not consistent with data based on protein coding loci. The six most informative SSR loci are unlinked and provide probabilities of single parent exclusion and genetic identity exceeding 0.99 and one in one million, respectively, both of which are reasonable standards for colony management purposes. PMID- 9209586 TI - Oral 18F-fluoro-2-deoxyglucose for primate PET studies without behavioral restraint: demonstration of principle. AB - We describe a method of orally administering 18F-fluoro-2-deoxyglucose (FDG) for positron emission tomography (PET) scans to determine local cerebral metabolic rates for glucose (LCMRGlc), normalized to that of whole brain, in fully conscious, non-restrained primates. Oral FDG-PET studies were performed in both non-restrained and chaired monkeys, and in one human where results could be compared with traditional intravenous FDG administration. The oral route of FDG administration gave images and whole brain-normalized PET LCMRGlc results comparable to those obtained by the intravenous route. This oral FDG-PET method may provide a useful means by which to obtain measures of LCMRGlcs for brain structures, relative to each other, in non-restrained, non-drugged primates in field and laboratory studies. This method might also have clinical applications for PET studies of children. PMID- 9209587 TI - Birth rate and mortality rate of infants with congenital malformations of the limbs in the Awajishima free-ranging group of Japanese monkeys (Macaca fuscata). AB - The birth rate and mortality rate of infants with congenital malformations of the limbs were examined in the Awajishima free-ranging group of Japanese macaques (Macaca fuscata). Of the 606 infants born between 1978 and 1995, 86 (14.2%) were malformed. The male-female ratio did not differ between malformed and normal infants. Most kin-groups included females who gave birth to malformed infants at least once. The mortality rate within the first year after birth for malformed infants (28.2%) was significantly higher than that for normal infants (10.0%). However, this indicates that more than 70% of malformed infants were able to survive for the first year of life, even though they were unable to cling to their mother's ventrum due to their limb deformities. This finding indicates that maternal care-taking is sufficient to enable malformed infants to survive during the early stages of development and that clinging by the infant is not necessary for the display of maternal care. PMID- 9209588 TI - Meiotic study of hybrids in the genus Eulemur and taxonomic considerations. AB - A reproductive study was conducted on seven hybrids of Eulemur showing chromosomal multivalents involving at least four chromosomes at the pachytene stage. Three individuals were infertile hybrids and one presented a reduced spermatogenesis. In three out of these four hybrids, multivalents were associated with the sex bivalent in a large number of spermatocytes (23%). The relative importance of the reduction of fertility in males linked to chromosomal multivalent formation as well as the genetic background is discussed with regard to the use of cytogenetic data for systematics. Our findings argue for the classification of Eulemur fulvus collaris and E. f. albocollaris in two separate species. In regard to their repartition area, their separation along a linear north-south axis in Madagascar is discussed. PMID- 9209589 TI - Evaluation of prognostic factors in endoscopic sinus surgery. AB - A retrospective analysis of 115 consecutive patients who underwent endoscopic sinus surgery for chronic sinusitis was conducted to evaluate the effect on outcome of variables including previous sinus surgery, allergy, asthma, and computed tomography stage of disease. Outcome was assessed in each patient by a survey in which the patient rated the benefit of surgery in terms of percent improvement in different symptoms compared with symptoms before surgery. Outcome was also assessed by the need for revision surgery and the presence of endoscopic criteria for failure. The results indicate that allergy and previous sinus surgery are associated with lower individual symptom scores, but no variable was associated with overall symptomatic failure. Previous sinus surgery was strongly associated with the need for subsequent revision surgery. The computed tomography stage was strongly associated with endoscopic evidence of failure. In conclusion, both history of previous sinus surgery and computed tomography stage of disease are correlated to poor outcomes after endoscopic sinus surgery. PMID- 9209590 TI - Correlation of clinical examination with computer tomography in paranasal sinus disease. AB - The accuracy of clinical examination in predicting radiologic paranasal sinus disease was investigated. Two hundred forty-seven patients who underwent endoscopic sinus surgery over a 4-year period were clinically and radiologically staged prior to surgery. A clinical staging system was developed utilizing fiberoptic intranasal examination to adequately visualize the anterior and posterior ostiomeatal complex structures with special attention to the mucosal status. The degree to which the clinical staging was able to predict and correlate with radiologic staging and predict the degree of sinus opacification was determined. Clinical and radiological presence of middle turbinate anomalies, septal deviation, and other structural anomalies were also evaluated. Clinical examination correlated well with radiologic examination: 74% sensitivity and 84% specificity). More than 94% of the patients with frank polyp disease had pansinus opacification involving the sphenoid. In the presence of normal mucous membranes, the absence of middle turbinate anomalies correlated with a normal computed tomography. However, with the exception of middle turbinate hypertrophy, the ability to clinically predict concha bullosa or paradoxical curvature was low and did not influence the overall computed tomography result. PMID- 9209591 TI - Improved video documentation of endoscopic sinus surgery made possible with desktop digital video. AB - Endoscopic cameras in endoscopic sinus surgery (ESS) allow for easy video documentation of surgery. This often leads to the build-up of extensive videocassette libraries where the prospective viewer is subjected to a lengthy and cumbersome videotape search before accessing a sequence of interest. Footage is difficult to use for educational purposes. New techniques for storing, retrieving, and presenting ESS video footage were developed. Desktop computer systems were used to digitize video footage. Short video sequences containing highlights of interesting ESS cases were prepared using video-editing programs. The resulting images were stored in individual segments on recordable CD-ROM disks. This technique was used to archive all video footage from our ESS procedures for a 6-month period. Archiving footage on CD-ROM rather than videotape has made the storage of large quantities of information possible while now allowing for the random access of segment(s) of interest. It has simplified preparation of video projects, reduced production time 10-fold and has eliminated the need for professional video-editing services. These techniques have also been adapted for the development of an educational CD-ROM demonstrating the component steps of ESS, which can be played back on any CD-ROM-equipped computer. PMID- 9209593 TI - Facial skeletal growth after endoscopic sinus surgery in the piglet model. AB - This study examined the effects of varying degrees of endoscopic sinus surgery on the growing midface and snout in pigs. In this randomized, controlled experiment, thirty 40-60 pound pigs were placed in five experimental groups: (1) unilateral uncinectomy; (2) bilateral uncinectomy; (3) unilateral uncinectomy, anterior ethmoidectomy, maxillary antrostomy; (4) bilateral uncinectomy, anterior ethmoidectomy, maxillary antrostomy; (5) unoperated controls. Animals were killed after 3 months and growth was assessed, according to linear and spatial measurements of multiple craniofacial regions. Euclidean distance matrix analysis showed significant restrictive shape alterations in a linear fashion in groups 1 4 (p < .05). These alterations occurred in the surgical field of the snout, midsnout, and maxilla. Endoscopic sinus surgery causes significant restrictive effects in the growing porcine facial skeleton. PMID- 9209592 TI - Development of the paranasal sinuses in children. AB - The development of computed tomography and functional endoscopic sinus surgery has improved diagnosis and management of sinusitis. It has also renewed interest in the developmental anatomy of the paranasal sinuses. There are significant differences between adult and pediatric sinus anatomy, and to safely perform functional endoscopic sinus surgery in children, the surgeon must be aware of these differences. To define the developmental anatomy of the paranasal sinuses, we analyzed 145 computed tomograms from patients under 18 years of age. The study emphasized landmarks at the level of the maxillary sinus ostium. In addition, distances and angles from the nasal spine to various points in the sinuses were determined. The structures were identified and traced on a digitizing tablet. Means and standard deviations were calculated for each measure as a function of age. This study can aid a better understanding of sinus development in children and provide guidance to the endoscopic sinus surgeon. PMID- 9209594 TI - The anatomic relevance of the Haller cell in sinusitis. AB - In current theories of sinusitis, obstruction at the ostiomeatal complex leads to localized inflammation and infection. Haller cells, an extension of ethmoid pneumatization along the maxillary antrum roof, have also been suggested as a causative factor in sinusitis because of their ability to cause narrowing of the infundibulum. Coronal CT scans were reviewed in 154 patients to evaluate the role of Haller cells in sinusitis. Haller cells were present in 34% of patients. The cells were graded as small, medium, or large, and correlated with radiologic evidence of sinusitis (e.g., mucosal thickening or opacification). A statistically significant increase in maxillary sinus mucosal disease was noted in patients with medium or large Haller cells (45.8%) versus those with small cells (28.9%, p < 0.05). Thus obstructive medium and large Haller cells may be an etiologic factor in sinusitis. PMID- 9209596 TI - The effects of gelatin film stents in the middle meatus. AB - Controversy exists regarding the management of the middle meatus after pediatric functional endoscopic sinus surgery (FESS). To prevent adhesions following pediatric FESS, gelatin film stenting of the middle meatus has been recommended. The effects of stenting, however, have not been established. Fifty-one children with similar degrees of bilateral sinus disease had a gelatin film stent placed in one middle meatus on completion of FESS, while the opposite meatus was not stented. Two to three weeks later at the time of a second, staged procedure, the sides were compared for the presence of the stent, adhesions, granulaion tissue, and patency of the maxillary sinus ostia. In 11 children the postoperative findings were more severe in the side without the stent, whereas in 29 children they were more severe in the stented side. There was no difference between the sides in 11 children. Although gelatin film stenting benefits some children, it should not be used routinely following pediatric FESS but should be reserved for children who are predisposed to develop adhesions or have poor prognostic factors, such as immunodeficiency and ciliary dyskinesia. PMID- 9209595 TI - Pyogenic granuloma--an uncommon complication of nasal packing. AB - Nasal packing is a very common procedure in the otolaryngologic service for nasal bleeding and postoperative hemostasis. However, a pyogenic granuloma complicated from nasal packing has not been reported in the literature. A 50-year-old man underwent nasal packing by use of vaseline gauze due to nasal bleeding. Two weeks later, a dark brown nasal tumor was found in his nasal cavity. He underwent partial turbinectomy for removal of the tumor. The pathology demonstrated a pyogenic granuloma. It is the first case of a confirmed pyogenic granuloma complicated from nasal packing in the literature. To prevent complications and decrease the discomfort, we strongly recommend the use of inflatable balloons or nontraumatic materials for nasal packing. PMID- 9209597 TI - Goblet cell density of the inferior turbinates in patients with perennial allergic and nonallergic rhinitis. AB - Nasal mucus production is regulated by submucosal glands and epithelial goblet cells. The role and especially the number of the latter are quite debatable. The present study compares the distribution and density of goblet cells in the inferior turbinates of patients with perennial allergic and nonallergic rhinitis to normal controls. The periodic acid Schiff-alcian blue whole-mount method was used to identify and count their number. Goblet cell distribution was found nonhomogeneous, and considerable variations were observed among adjacent localities of the same specimen. The mean number of goblet cells per mm2 was 6499 in normal controls (n = 12), 6818 in patients with perennial allergic rhinitis (n = 13), and 6801 in patients with perennial nonallergic rhinitis (n = 18). Statistical analysis confirmed that the density of goblet cells did not differ significantly between patients with and without allergy, as well as between each group of patients and controls. Therefore, it could be concluded that the number of goblet cells in the inferior turbinate is not influenced by the presence of perennial allergic or nonallergic rhinitis. PMID- 9209598 TI - Effects of anti-allergic drugs on substance P (SP) and vasoactive intestinal peptide (VIP) in nasal secretions. AB - To clarify the effects of anti-allergic drugs on substance P (SP) and vasoactive intestinal peptide (VIP) levels in nasal secretions, we employed competitive enzyme-linked immunoassays to measure concentrations of those neuropeptides in nasal secretions from 40 patients with house dust nasal allergy before and after administration of azelastine and oxatomide. One mg of azelastine and 30 mg of oxatomide were administrated twice a day for 4 weeks. Mean values of SP concentrations and ratios of SP to total protein of the nasal allergy group were significantly higher than those of the control group (p < 0.002). The VIP/total protein ratio of the allergy group was also significantly higher than that of the control group, although the VIP concentration alone was not. Mean levels of SP and VIP from patients with severe symptoms were significantly higher than those of the control group (p < 0.05), although those values were not significantly different between patients with moderate symptoms and control subjects. Azelastine and oxatomide effectively reduced SP levels in nasal secretions (p < 0.005), but they did not significantly decrease VIP levels. The reduction of SP levels was significant in patients with excellent responses to those drugs (p < 0.005), but not in patients with poor responses. These findings suggest that SP and VIP levels in nasal secretions may reflect the clinical state of nasal allergy and be one of the better parameters available for evaluating the clinical efficacy of anti-allergic drugs against nasal allergy. PMID- 9209599 TI - Ether inhaler, 1847. PMID- 9209600 TI - The history of anaesthesia in Australia--150 years. PMID- 9209601 TI - Severe community-acquired pneumonia. PMID- 9209602 TI - Barriers to hepatitis C transmission within breathing systems: efficacy of a pleated hydrophobic filter. AB - It has been suggested that breathing circuits contaminated with body fluids may provide a route of nosocomial patient-to-patient transmission of the hepatitis C virus. Thus, a number of authorities have recommended the use of breathing circuit filters to minimize such risks. The present study sought to simulate a humidified breathing circuit and evaluate two different designs of breathing circuit filters to determine their efficacy in preventing passage of the hepatitis C virus. A hydrophobic pleated-membrane filter consistently prevented the passage of hepatitis C virus while a large-pore "electret" filter design was ineffective. We conclude that not all filter types are equally suited to preventing the passage of viruses and we therefore consider it essential that, if filters are intended to prevent the passage of named pathogens in a humidified breathing circuit, they should be evaluated in a similar experimental system to that described in order to prove their efficacy. PMID- 9209603 TI - Bacteraemia during direct laryngoscopy and endotracheal intubation: a study using a multiple culture, large volume technique. AB - Bacteraemia secondary to orotracheal intubation has been reported to occur in 0 5.3% of patients. Bacteraemia detection is dependent upon several factors including the volume of blood per culture and the number of cultures. Prior studies used small volumes of blood and one or two cultures, and may therefore have underestimated the incidence of bacteraemia. Sixty-two adult patients who underwent direct laryngoscopy and endotracheal intubation were studied. Baseline blood cultures were sterile in all patients. After intubation, four blood cultures were obtained in ten minutes, with 10 ml being evenly divided between aerobic and anaerobic media. Two patients (3.2%) became bacteraemic. This is a lower incidence than occurs in association with other procedures for which The American Heart Association does not recommend administration of prophylactic antibiotics. Therefore, prophylactic antibiotics are not recommended prior to direct laryngoscopy. However, when a prophylactic antibiotic is administered prior to surgery, it would be best to administer the antibiotic prior to direct laryngoscopy and intubation. PMID- 9209604 TI - Amrinone versus dobutamine in cardiac surgical patients with severe pulmonary hypertension after cardiopulmonary bypass: a prospective, randomized double blinded trial. AB - We compared the relative effects of dobutamine (5 micrograms/kg/min) and amrinone (1.0 mg/kg bolus followed by 10 micrograms/kg/min) on right and left ventricular function and pulmonary arterial pressures during weaning from cardiopulmonary bypass in patients with a mean preoperative pulmonary pressure > 30 mmHg. Twenty patients scheduled for mitral valve replacement were studied in a prospective, randomized, double-blind trial. Patients receiving amrinone had a greater increase in cardiac index (CI) of 1.38 (+/-0.95) litre/min/m2 at separation vs 0.69 (+/-0.63) litre/min/m2 in the dobutamine group (P < 0.05). The amrinone group also had a greater increase in right ventricular ejection fraction (0.15 +/ 0.08 at separation from cardiopulmonary bypass versus an increase of 0.04 +/- 0.11 in those receiving dobutamine; P < 0.005). Amrinone produced a larger decrease in pulmonary artery wedge pressure 8.0 (+/-4.4) mmHg vs 0.75 (+/-6.6) mmHg at separation; pulmonary artery systolic and diastolic pressures also were reduced more in the amrinone group. There were no differences in heart rate, mean arterial pressure, central venous pressure and right ventricular stroke work index between patient groups. In the doses chosen, the use of amrinone compared to dobutamine was associated with a reduction in pulmonary arterial pressures and an increase in cardiac index and right ventricular ejection fraction after separation from bypass in patients with severe preoperative pulmonary hypertension. PMID- 9209605 TI - Estimation of cardiac output by noninvasive echocardiographic techniques in the critically ill subject. AB - We evaluated the accuracy of cardiac output estimations by three transthoracic echocardiographic techniques in critically ill subjects. This study was a prospective comparison study carried out in a general intensive care unit of a teaching hospital. The subjects had a broad range of diagnoses including pulmonary embolus, cardiogenic shock, septic shock, Legionnaire's disease and perioperative myocardial infarction. All patients requiring pulmonary artery catheterization underwent echocardiographic cardiac assessment with comparison of findings to those obtained by thermodilution techniques. Nineteen studies on eighteen patients were performed, with cardiac output calculated by the two chamber Simpson's, four-chamber Simpson's, and left ventricular outflow tract (LVOT) Doppler methods. Acceptable data was obtained in those patients without mitral regurgitation. There was good correlation between the thermodilution technique and Simpson's two-chamber method (r = 0.91), but less so with the Simpson's four-chamber method (r = 0.77). All studies were included in the LVOT Doppler method with a good correlation (r = 0.94). A plot of differences between methods using the Bland and Altman statistical method indicated that only the LVOT Doppler method demonstrated acceptable agreement with a mean of 0.2 litres/minute, standard deviation of 0.82 litres/minute and 95% limits of agreement of -1.5 to +1.9 litres/minute. We concluded that the LVOT Doppler method was the only one which demonstrated acceptable agreement between the thermodilution method and echocardiographic techniques in all critically ill patients studied. PMID- 9209606 TI - Adverse cardiovascular effects of ketamine infusion in patients with catecholamine-dependent heart failure. AB - The longterm effects of ketamine on haemodynamic parameters and exogenous catecholamine requirements were studied in twenty-five critically ill patients with catecholamine-dependent heart failure. Following sedation with midazolam (0.15 +/- 0.07, mg.kg-1.h-1) and sufentanil (0.88 +/- 0.33 microgram.kg-1.h-1), patients with impaired left ventricular function (left ventricular ejection fraction area 30 +/- 7%) were randomly assigned to receive ketamine (2.5 +/- 0.9 mg.kg-1.h-1) and midazolam (Group A) or remained on sufentanil/midazolam (Group B). Haemodynamic measurements were performed throughout the first 24 hours after randomization. In group A cardiac index decreased by 21% (P = 0.01), mean arterial pressure increased by 13% (P = 0.01), mean pulmonary artery pressure by 14% (P = 0.04), pulmonary capillary wedge pressure by 20% (P = 0.03), and systemic vascular resistance index by 38% (P < 0.001). No significant cardiovascular effects were observed in Group B. Neither group had significant changes of exogenous catecholamine requirement. In conclusion, ketamine exhibits potential negative cardiovascular effects in patients with catecholamine dependent heart failure. Therefore, ketamine should not be considered a first line drug for longterm sedation of patients with impaired left ventricular function. PMID- 9209607 TI - Evaluation of the onset time and intubation conditions of rocuronium bromide in children. AB - We have assessed, in children aged three to eight years, the intubating conditions after administration of rocuronium 0.6 mg/kg at 50 or 60 seconds, in groups of 15 patients. Intubating conditions were excellent in 11, good in 3 and fair in 1 patient at 50 seconds and excellent in 12 and good in 3 patients at 60 seconds. The mean onset time, for all patients, to when the first twitch of the train of four (T1), measured at the adductor pollicis, was depressed to less than 30% and 5% of control was 50 (SD 11.4) seconds and 94 (SD 31.7) seconds respectively. Depression of T1 to less than 30% of control, measured at the adductor pollicis in children, appears to indicate that intubating conditions will be clinically acceptable when using rocuronium. PMID- 9209609 TI - Plasma somatostatin correlates with blunted thyrotropin secretion after stimulation by thyrotropin-releasing hormone in critical illness. AB - To clarify whether plasma somatostatin affects thyrotropin secretion in critical illness, plasma somatostatin and thyrotropin responses to thyrotropin-releasing hormone were studied in forty-three critically ill patients. High somatostatin levels were associated with blunted thyrotropin secretion in critically ill patients. There was an inverse correlation between plasma somatostatin levels and the maximum increment of thyrotropin after stimulation by thyrotropin-releasing hormone. Decreased somatostatin and increased thyrotropin secretion before discharge from the intensive care unit were demonstrated in survivors. On the other hand, non-survivors maintained high somatostatin levels and had blunted thyrotropin secretion during their intensive care admission. These results suggest that high plasma somatostatin levels may play a role in the blunted thyrotropin secretion observed in critical illness. PMID- 9209608 TI - Comparison of 0.5% ropivacaine and 0.5% bupivacaine in lumbar epidural anaesthesia for lower limb orthopaedic surgery. AB - The purpose of this study was to compare the epidural use of 0.5% ropivacaine and 0.5% bupivacaine in patients undergoing lower limb orthopaedic surgery. In a double-blind, randomized, multi-centre study involving 67 patients, thirty-two patients received 20 ml of 0.5% ropivacaine and 35 patients received 20 ml of 0.5% bupivacaine at the L2,3 or L3,4 interspace. Parameters measured were the onset time, duration and spread of sensory block, the onset time, duration and degree of motor block, the quality of anaesthesia and the heart rate and blood pressure profile during block onset. Four patients (3 ropivacaine, 1 bupivacaine) were excluded from the study due to technical failure of the block. The onset and duration of analgesia at the T10 dermatome (median, interquartile range) was 10 (5-15) minutes and 3.5 (2.7-4.3) hours respectively for ropivacaine, and was 10 (6-15) minutes and 3.4 (2.5-3.8) hours respectively for bupivacaine. Maximum block height (median, range) was T6 (T2-T12) for ropivacaine and T6 (C7-T10) for bupivacaine. Nine patients receiving ropivacaine and eight patients receiving bupivacaine developed no apparent motor block. The incidence of complete motor block (Bromage grade 3) was low in both groups, being 4/27 for ropivacaine and 6/34 for bupivacaine. In the ropivacaine group, motor and sensory block were judged to be satisfactory in 78% of patients. In the bupivacaine group, motor and sensory block were judged to be satisfactory in 71% and 62% of patients respectively. Cardiovascular changes were similar in both groups. No statistical differences were found between the two groups regarding any of the study parameters. PMID- 9209610 TI - Pharmacokinetics and pharmacodynamics of suxamethonium. AB - A study was undertaken to determine the time constants of elimination and effect compartment equilibration of suxamethonium and for the slope exponent of the Hill equation. Twelve patients were anaesthetized with thiopentone, fentanyl, and isoflurane in nitrous oxide and oxygen. After allowing conditions to become stable, they were administered three small doses of suxamethonium by rapid intravenous injection. The responses to supramaximal stimulation of the ulnar nerve were recorded by EMG in one and by accelerometry in eleven subjects. Because of failure to recover to control conditions, one subject was deleted from analysis. The recorded drug effect was used in a non-linear curve fitting technique to derive estimates of the pharmacokinetic and pharmacodynamic parameters. The plasma concentration of suxamethonium was adequately represented by a single compartment model. The mean half-life of elimination was 47 s with a 95% confidence interval of 24 to 70 s; that of effect compartment equilibration, 211 s with a 95% confidence range of 139 to 282 s. The average slope exponent was 6.4 and its 95% confidence range was 4.6 to 8.2. The data from the first two doses were used to predict the time taken for the third dose to recover 50%. The predictions showed a mean bias of < 1% (NS) and a mean error of 21%, with a confidence interval of 5 to 37%. In only two out of eleven subjects was the prediction error greater than 30%. PMID- 9209611 TI - A normal distribution model for the pharmacodynamics of nondepolarizing neuromuscular blockers. PMID- 9209612 TI - National audit of anaesthesia for major abdominal surgery. PMID- 9209613 TI - An early anaesthetic in Papua New Guinea. AB - A search for information about early anaesthetics administered in Papua New Guinea has revealed that an ether or chloroform anaesthetic was given, probably for a retained placenta, at Port Hunter on December 9, 1880. The anaesthetist or anaesthetic assistant was the Reverend George Brown, a Wesleyan Methodist missionary. PMID- 9209614 TI - Nathan P. Rice and Trials of a Public Benefactor, 1859--historical notes on the facsimile of 1995. PMID- 9209615 TI - Peripartum cardiomyopathy: four case histories and a commentary on anaesthetic management. PMID- 9209616 TI - Back pain following postoperative epidural analgesia: an indicator of possible spinal infection. PMID- 9209617 TI - Latex allergy during anaesthesia: cautionary tales. PMID- 9209618 TI - The Geoffrey Kaye Oration. ASA/ANZCA combined scientific meeting, Perth, W.A. October 30, 1996. PMID- 9209619 TI - Narkomed 4E power failure. PMID- 9209620 TI - Biting on ET tubes. PMID- 9209621 TI - Modification of the Benjamin Jet Tube. PMID- 9209623 TI - Failed spinal anaesthesia in combined spinal-epidural anaesthesia. PMID- 9209622 TI - Intraoperative bronchoscopy during thoracotomy through an LMA in an infant. PMID- 9209624 TI - Transjugular intrahepatic portosystemic shunt (TIPS) complicated by complete heart block. PMID- 9209625 TI - Neuromuscular block. PMID- 9209626 TI - Wet lungs and impedance cardiography. PMID- 9209627 TI - Awake blind oral intubation using a malleable stylet. PMID- 9209628 TI - Awake oral breath-guided intubation. PMID- 9209629 TI - Recovery pain protocols. PMID- 9209630 TI - Fractured 27 gauge Whitacre spinal needle. PMID- 9209631 TI - Erythromycin--a pro-kinetic and pro-arrhythmogenic agent. PMID- 9209632 TI - The problem of classifying lymphomas: an orderly prescription for progress. AB - In the late 1960s and early 1970s, the most widely recognized 'new' classifications of the non-Hodgkin's lymphomas were those proposed by Rappaport (the 'Rappaport' classification) and by Lennert (the 'Kiel' classification). With the advent of immunologic and histochemical markers in the early 1970s, however, new concepts arose to supplement the traditional purely morphologic approach to diagnosis and classification of these tumors. Lymphomas were increasingly recognized to be neoplasms of the immune system, composed of malignant proliferations which retained many of the morphologic and functional characteristics of their normal counterparts. These advances led to a flurry of new classifications proposed in 1974-1976, leading to confusion for both clinicians and pathologists, perhaps most evident at the International Cancer Congress in Florence in 1974. To address this problem, the National Cancer Institute (USA) sponsored an international workshop of expert pathologists and clinicians on 4-5 September 1975. It became apparent at that meeting that only a well-planned retrospective study would provide data for meaningful progress and resolution of differences. From 1976 to 1980, such a massive collaborative project was accomplished and served as the basis for the Working Formulation for Clinical Usage, proposed as a vehicle for translation among the six tested schema. Since the Working Formulation was published in 1982 there have been momentous strides in scientific and clinical understanding of these cancers, fueled by contributions from immunology, cytogenetics, and molecular biology. To recognize and disseminate understanding of these newer observations, the International Lymphoma Study Group promulgated in 1994 a new proposal entitled 'A Revised European-American Classification of Lymphoid Neoplasms'. As a sequel to another international assembly of pathologists and clinicians, held at the National Cancer Institute (USA) on 21-23 March 1994, a second large-scale retrospective study has been accomplished, the results of which were presented at the Sixth International Conference on Malignant Lymphoma, 5-8 June 1996, along with data from other institutions and cooperative groups. Concurrent with these events, the World Health Organization has enlisted a committee of expert pathologists to prepare a new edition of 'Neoplastic Diseases of Hematopoietic and Lymphoid Tissues'. Composed of 10 pathology subcommittees and a clinical advisory committee, with broad international representation, this body should generate in the near future a consensus proposal with broad scientific and geographic support. These historical and ongoing efforts in lymphoma pathology are a paradigm for progress in clinicopathologic understanding of all cancers. PMID- 9209633 TI - Principles of the revised European-American Lymphoma Classification (from the International Lymphoma Study Group). AB - The International Lymphoma Study Group has proposed a consensus classification for lymphoid neoplasms. Lymphoid neoplasms are defined as distinct biological entities, based on a combination of morphologic, immunophenotypic, genetic, and clinical features. Each distinct disease may have a range of histologic grade and clinical aggressiveness. Although many distinct diseases can now be recognized, three of them (follicular lymphoma, diffuse large B-cell lymphoma, and Hodgkin's disease) account for the majority of the cases seen in Europe and the USA. Recognition of distinct disease entities is essential in order to develop and test effective therapies. PMID- 9209635 TI - CB/CC diffuse lymphoma: a distinct subtype of non-Hodgkin's lymphoma? A study of 1593 patients from a Danish population-based registry. Danish LYFO Study Group. AB - Between 1983 and 1993, 3165 cases of non-Hodgkin's lymphoma (NHL) were reported to the West Danish Lymphoma Registry (LYFO). Out of these, 148 (4.7%) were of the CB/CC diffuse subtype according to the Kiel classification. However, in the new European-American NHL consensus classification (REAL, 1994), CB/CC diffuse lymphoma was categorized as a provisional subtype only. In the LYFO material, death-probability curves show a significantly shorter survival in CB/CC diffuse than in CB/CC follicular. In order to detect further possible differences between CB/CC diffuse and other NHL subtypes, a number of clinical parameters at presentation were analyzed in a subset of five types of lymphoma. This subset included 148 cases of CB/CC diffuse, 435 cases of CB/CC follicular, 667 cases of CB diffuse, 202 cases of CC diffuse, and 131 cases of peripheral T-cell lymphoma. Using logistic regression analysis, significant differences could be demonstrated between CB/CC diffuse and the four other subtypes as regards sex ratio, age distribution, and sites of both nodal and extranodal involvement. These findings indicate that CB/CC diffuse has a distinct clinical phenotype and imply the existence of real biological differences between CB/CC diffuse and other subtypes of lymphoma. PMID- 9209634 TI - Classification of T-cell and NK-cell neoplasms based on the REAL classification. AB - Mature or peripheral T-cell lymphomas are uncommon, accounting for only 10%-15% of all non-Hodgkin's lymphomas. The classification of these neoplasms has been controversial. In contrast to B-cell lymphomas, cytologic grade has not been very useful in predicting the clinical course. This finding may result from the generally aggressive clinical course associated with T-cell lymphomas. Prior studies have suggested that stage of disease may be more important than cytologic subtype. Clinical presentation is very important in the classification of T-cell malignancies. For T-cell lymphomas, cytologic features alone are not sufficient to distinguish among disease entities. For example, adult T-cell leukemia/lymphoma (ATLL) often cannot be distinguished morphologically from HTLV 1-negative T-cell lymphomas. Most extranodal T-cell lymphomas appear to be derived from cytotoxic T cells, which express perforin, TIA-1, and granzyme B. Three broad groups of T-cell malignancies can be identified: (1) leukemic or systemic disease; (2) nodal disease; (3) extranodal disease. Anaplastic large cell lymphoma (ALCL) is probably the single most common subtype of T-cell lymphoma. Classical ALCL should be distinguished from primary cutaneous ALCL (CD30+ lymphoproliferative disease of the skin), which is a distinct disease entity. PMID- 9209636 TI - Primary B-cell lymphomas of the skin. AB - Primary B-cell lymphomas of the skin are not as rare as is generally believed. They are defined as malignant B-cell proliferations presenting with cutaneous involvement alone and no evidence of extracutaneous manifestations over a period of at least six months when complete staging has been performed. The major subtypes are follicle center-cell lymphoma of the head and trunk, immunocytoma and large B-cell lymphoma of the leg (EORTC classification 1996). Also of interest is the recently recognized marginal-zone B-cell lymphoma. Primary B-cell lymphomas of the skin differ significantly from nodal lymphomas. Awareness of their special clinical behavior should prevent unnecessarily aggressive treatment. PMID- 9209637 TI - Specific detection of Helicobacter pylori and non-Helicobacter pylori flora in small- and large-cell primary gastric B-cell non-Hodgkin's lymphoma. AB - BACKGROUND: Primary gastric non-Hodgkin's lymphomas possibly develop in response to local infection by Helicobacter pylori (H. pylori). We investigated the presence of H. pylori and non-H. pylori flora histologically in small- and large cell primary gastric lymphoma using a specific staining method. MATERIALS AND METHODS: Specimens of 52 cases of primary gastric lymphoma (17 small cell, 35 large cell) were stained with modified Giemsa (MG) and immunohistochemically using a polyclonal antibody against H. pylori (IHC). RESULTS: Thirty-two cases (61.5%) (small cell 76% versus large cell 53%, P > 0.05) showed immunoreactivity for H. pylori in the mucosa surrounding the tumor. Remarkably, there was localization of H. pylori in the neck of the gastric glands in 3 cases. Non-H. pylori flora was seen in 35 cases (76.3%) (small cell 53% versus large cell 74%, P > 0.05). In 20 cases, this non-H. pylori flora was mixed with H. pylori. Five cases showed no bacterial flora at all. CONCLUSIONS: (1) Using immunohistochemistry, the prevalence of gastric lymphoma cases with H. pylori (61.5%) approximates that of H. pylori in the normal population. (2) No statistical difference was found between the occurrence of H. pylori and non-H. pylori bacterial flora in small- versus large-cell lymphoma. (3) Our results suggest that H. pylori may not be the only etiologic factor in primary gastric lymphoma. PMID- 9209639 TI - Pathogenesis of gastric lymphoma: the enigma in Hong Kong. AB - BACKGROUND: Helicobacter pylori (H. pylori) infection has been postulated to be a pathogenetic factor in gastric lymphoma. However, the etiological factors for gastric lymphoma could vary in different populations. MATERIALS AND METHODS: We looked for histological evidence of H. pylori infection in 53 gastrectomy specimens from Hong Kong Chinese patients with primary gastric B-lymphoma. We also screened for Epstein-Barr virus (EBV) in these cases using in situ hybridization with oligonucleotide probes for EBV-encoded small RNA1 and 2. RESULTS: H. pylori was found in 29 of 53 (55%), including 8 of 13 (62%) cases of low-grade lymphoma of mucosa-associated lymphoid tissue (MALT) type. These infection rates in gastric lymphoma are lower than those reported in Western populations (80%-100%) and comparable to that found in healthy Chinese blood donors (55%) or in non-ulcer dyspeptic patients (52%-57%). EBV was found in tumor cells only in one case of high-grade gastric lymphoma with low-grade MALT component which was H. pylori-negative, and in occasional nontumor-lymphoid cells in 7 other cases. CONCLUSIONS: These results suggest that (1) the role of H. pylori in pathogenesis of gastric lymphoma may vary in different populations; (2) very few gastric lymphomas are associated with EBV; (3) not all low-grade gastric MALT lymphomas are H. pylori-dependent. PMID- 9209638 TI - Helicobacter pylori eradication for the treatment of low-grade gastric MALT lymphoma: follow-up together with sequential molecular studies. AB - BACKGROUND: Helicobacter pylori infection is associated with low-grade gastric MALT lymphoma, and available data support that the eradication of the H. pylori can cause histological regression of the lymphoma. PATIENTS AND METHODS: Nine patients with low-grade gastric MALT lymphoma were treated with amoxicillin, metronidazole, and omeprazole for 14 days in a prospective study. Patients were followed up with sequential endoscopy, mapping gastric biopsies, and molecular studies with PCR amplification of the IgH gene in order to assess the response to H. pylori eradication and the evolution of the histological molecular responses. RESULTS: H. pylori was eradicated in all patients and reinfections were not demonstrated. After H. pylori eradication treatment, the lymphoma regressed both endoscopically and histologically in eight of the nine patients (88.8%). In four of the eight histologically cured patients, no clonal band was detected by PCR; in the remaining four patients; PCR identified a clonal band, which disappeared in all patients after a mean of 12 +/- 4 months. No clonal band was detected by the PCR analysis in any of the eight patients with histological regression after a median of 7 +/- 6 months (range 1-20). The seven followed-up patients have a persistent clinical and histological remission after a median of 14 +/- 5 months. CONCLUSIONS: (1) Low-grade gastric MALT lymphoma can be histologically cured with eradication therapy for H. pylori. (2) After histological regression, PCR amplification of the IgH gene can identify an eventually persisting clonal population. (3) Sequential histological and molecular studies are essential for the assessment of the evolution of the lymphoma. (4) The clonal population tends to disappear, but its disappearance may be delayed for months. (5) Patients with histological regression but with a persistent clonal band should not be treated unless the lymphoma can be histologically demonstrated. All these observations suggest that gastric lymphoma can be effectively cured, but the ultimate fate of these patients is unknown until long-term follow-up studies are available. PMID- 9209640 TI - Trans-rearrangements and the risk of lymphoid malignancy. AB - BACKGROUND: Antigen receptor 'trans-rearrangements' occur in all individuals and represent a particular type of genetic instability whose mechanism, V(D)J recombination, is the same as that required for the development of a normal immune response. DESIGN: We have measured the level of trans-rearrangements in a variety of populations characterized by increased risk for the development of lymphoid malignancy. The human populations studied include those with an inherited predisposition to lymphomagenesis (ataxia-telangiectasia patients), as well as populations at increased risk because of an occupational (agriculture workers) or iatrogenic (Hodgkin's disease patients) exposure. In addition, we have developed a mouse model for the more controlled analysis of these events. RESULTS: There is a correlation between the absolute number of trans rearrangements (as a population mean or median) and risk of lymphoma, whether that risk is based on an inherited predisposition or acquired exposure. CONCLUSION: This assay may serve as an easily measurable biomarker of lymphoma risk. If so, it is more than a fortuitous biomarker since the same mechanism responsible for the formation of trans-rearrangement is, at least in part, responsible for the majority of presumably 'malevolent' translocations associated with the transformation of lymphocytes. PMID- 9209641 TI - Changing trends in the incidence of non-Hodgkin's lymphoma in Europe. Biomed Study Group. AB - Non-Hodgkin's lymphoma (NHL) is not a uniform disease entity, and in order to investigate the reported changes in incidence we have set up a study in seven population-based cancer registries in Europe. The study is designed to look at changes in the incidence of total NHL and disease subgroups using standard definitions and methodology. The registries are based in Leeds, Dijon, Kuopio, Odense, Florence, Eindhoven, and Ragussa. The classification system we have used is based on the REAL classification and has utility for epidemiological studies. We have used it to convert data sets which have utilized both local cases and the ICD-O classification. In order to improve data reproducibility, CLL/LL, myeloma/MGUS, lymphoblastic disease, and Hodgkin's disease have been excluded because of the difficulty in defining incident cases accurately. The preliminary results of this study show that there is still an upward trend in incidence rate and that in Yorkshire this is 3% per annum in total NHL. The subgroups which are increasing are extranodal and nodal peripheral T-cell lymphoma. Similar increases in incidence have been reported for the other registries. We conclude that there is a continued upward trend in incidence of NHL, the causes of which are uncertain. PMID- 9209642 TI - Detection of trisomy 12 in CD34+ progenitor cells in a patient with B-cell chronic lymphocytic leukemia by fluorescence in situ hybridization. AB - B-cell chronic lymphocytic leukemia (B-CLL) is a slowly progressive disease resulting from the clonal expansion of mature B lymphocytes. The most frequent chromosomal abnormality is trisomy 12. Recently more aggressive therapeutic approaches using myeloablative therapy and autologous stem-cell support have been developed. Phase I/II studies have resulted in molecular remission and prolonged survival. One cause of relapse may be tumor-cell contamination of the transplant. We asked whether immunophenotypically identified hematopoietic progenitor cells are part of the malignant cell population in B-CLL. In a patient with trisomy 12, two subpopulations of hematopoietic progenitor cells, CD34+/CD38+ and CD34+/CD38- cells, were isolated by fluorescence-activated-cell-sorting; the sort purity was 98%. Trisomy 12 was detected in 13% of CD34+/38+ cells and in 34% of CD34+/38- cells. These data suggest that CD34+ cells are involved in the malignant clone of patients with B-CLL. The results are of significance for future strategies using autologous stem-cell transfusion. PMID- 9209643 TI - Long-range amplification of genomic DNA detects the t(2;5)(p23;q35) in anaplastic large-cell lymphoma, but not in other non-Hodgkin's lymphomas, Hodgkin's disease, or lymphomatoid papulosis. AB - DESIGN: Determine the frequency of t(2;5)(p23;q35) in anaplastic large-cell lymphoma (ALCL), non-Hodgkin's lymphoma (NHL), Hodgkin's disease (HD), and lymphomatoid papulosis (LP). PATIENTS AND METHODS: The t(2;5) was detected with a long-range nested polymerase chain reaction (PCR) using 0.5 microgram of DNA (60,000-80,000 cells), 5'-primers from the NPM gene, 3'-primers from the ALK gene, agarose electrophoresis, hybridization, and autoradiography. Patients were evaluable if a 3016 base pair amplicon could be generated from tumor DNA with beta-globin primers. RESULTS: Amplicons were detected by PCR of genomic DNA from three ALCL cell lines and four primary ALCLs known to t(2;5) positive. DNA from t(2;5)-positive cell lines diluted 10(4)-fold or 10(5)-fold generated amplicons in 100% or 20% of reactions, respectively. Archival tumor DNA from 144 patients was amplifiable by beta-globin amplicons in 126 (88%) who are considered evaluable for this study. Twenty-two had ALCL, 69 other NHLs, 30 HD, and five LP. Genomic DNA PCR detected the t(2;5) in 5 of 22 with ALCL (23%, 95% confidence intervals [95% CI] 8%-45%) but not in those with NHLs, HD, or LP. Among ALCLs the t(2;5) was confined to 5 of 20 with nodal presentations (25%, 95% CI 9%-49%), among whom it was seen in 5 of 15 with T-cell or null-cell phenotype (33%, 95% CI 12%-62%), in 4 of 11 with age < 40 years (36%, 95% CI 11%-69%), and in 4 of 9 with nodal presentations, T-cell or null-cell phenotype, and age < 40 years (44%, 95% CI 14%-79%). Amplicon sizes were different between cell lines and patients, reflecting unique genomic DNA breakpoints, as confirmed by DNA sequencing, and served as an internal control against specimen cross-contamination in the laboratory. CONCLUSIONS: Long-range PCR of genomic DNA detects t(2;5) only in ALCL, but not in other NHLs, HD, or LP. Long-range PCR may be useful in establishing diagnosis, determining prognosis, and monitoring minimal residual disease in ALCL. PMID- 9209644 TI - The use of fluorescent in situ hybridization for detection of the t(2;5)(p23;q35) translocation in anaplastic large-cell lymphoma. AB - BACKGROUND: Anaplastic large-cell lymphoma (ALCL) is a recently recognized, distinctive type of non-Hodgkin's lymphoma characterized by anaplastic large-cell cytology and expression of a member of the TNF-receptor family CD30. A characteristic chromosomal translocation has been identified in ALCL of T-or null cell lineage which juxtaposes a novel tyrosine kinase (anaplastic lymphoma kinase, ALK) located at 2p23 with the nucleophosmin gene (NPM) at 5q35. A chimeric mRNA transcript is produced, and the translocation results in constitutive expression of a truncated form of the ALK protein, p80. There is controversy concerning whether or not the translocation occurs in Hodgkin's disease. The aim of this study was to develop a methodology for fluorescent in situ hybridization (FISH) to detect the t(2;5)(p23;q35), and to compare the results with conventional cytogenetics, reverse-transcriptase PCR and immunostaining for the p80 protein. PATIENTS AND METHODS: Twenty-five cases of malignant lymphoma (11 ALCL and 14 HD) were studied. Immunohistochemistry was performed to confirm the diagnosis and for analysis of p80 expression. Conventional cytogenetics were analyzed on G-banded metaphase spreads. FISH was performed using whole chromosome paints for chromosomes 2 and 5 on metaphase spreads and YAC probes for interphase nuclei. Reverse-transcriptase PCR using primers for ALK and NPM was used to amplify the translocation breakpoint in extracted mRNA. RESULTS: Among 11 cases of ALCL examined by FISH, the translocation was detected in 4. Two of these cases also had RT-PCR and p80 staining performed, with positive results. Among 7 cases where the t(2;5) was not detected by FISH, 3 cases were examined by RT-PCR with negative results and 4 cases by p80 staining, also negative. The RT-PCR was negative in all 14 cases of Hodgkin's disease, 4 of which were also examined by FISH and found to be negative. CONCLUSION: Fluorescent in situ hybridization is a useful method for detection of the t(2;5)(p23;q35) in anaplastic large-cell lymphoma. The results concur with those of RT-PCR for the chimeric transcript and immunostaining for the p80 protein. The frequency with which the translocation was found was 36% in this small series, and no evidence of the translocation was found in cases of Hodgkin's disease. PMID- 9209645 TI - Analysis of the cyclin-dependent kinase inhibitors p18 and p19 in mantle-cell lymphoma and chronic lymphocytic leukemia. AB - BACKGROUND: Mantle-cell lymphoma (MCL) is characterized by overexpression of the G1 cyclin, cyclin D1, strongly implicating this cell-cycle regulatory element in MCL pathogenesis. Recently, loss-of-function mutations in cell-cycle negative regulatory elements, including p53 point mutations and deletions of the cyclin dependent kinase inhibitors (CDKI) p15 and p16 have been described in a subset of MCLs and have been associated with aggressive clinical course, blastic morphology, and extranodal dissemination. The objective of the present study was to analyze two newly identified members of the p16 (INK4A; MTS1) CDKI family, p18 and p19, in MCL. Such analyses have not been previously reported. PATIENTS AND METHODS: DNA was isolated from tissue biopsies, peripheral blood cells, or bone marrow cells of 45 patients with MCL and 15 with chronic lymphocytic leukemia (CLL). Southern blot analysis was performed with p18 and p19 probes and compared to placental control DNA and to control probe hybridizations for evidence of p18 or p19 gene deletion or rearrangement. RESULTS: P18 deletion was identified in one MCL but in no case of CLL. One MCL sample had rearrangement of the p18 gene; this case also had coexisting homozygous p15 and p16 deletion. Both cases with p18 abnormalities had blastic morphology, and one had extranodal disease with renal parenchymal invasion. CONCLUSIONS: P18 rearrangement or deletion as detected by Southern blot is a rare event in MCL, but may be associated with blastic morphology. P53 mutations and deletions of the CDKI p15 and p16 appear to be more frequent in MCL, although further studies are necessary to assess the presence of inactivating point mutations or altered expression of p16 family proteins. PMID- 9209646 TI - New knowledge about T-cell cytotoxicity. PMID- 9209647 TI - Normal and malignant B-cell development with special reference to Hodgkin's disease. AB - During their development, B lymphocytes are repeatedly selected for the expression of an appropriate surface receptor: the pre-B-cell receptor at the pre B-cell stage and surface immunoglobulin (Ig) at the transition from a pre-B cell to a mature B cell. Furthermore, stringent selection for B cells expressing high affinity antibodies operates when antigen-activated B cells proliferate within germinal centers (GC). Here, somatic point mutations are introduced into rearranged V region genes at a high rate, and B cells acquiring favorable mutations are selected to differentiate into memory B cells or plasma cells. In the frame of this developmental scheme, extending a recent analysis, we investigated 10 primary cases of Hodgkin's disease (HD) for B-lineage origin and clonality [1]. Single Hodgkin and Reed-Sternberg (H-RS) cells were micromanipulated from frozen tissue sections and analyzed by PCR for rearranged V genes. Clonal VH and/or V kappa/ V delta gene rearrangements were obtained from 9 of the cases. This shows that H-RS cells represent a clonal, B-lineage-derived population of tumor cells. Somatic mutations were found in all clonal VH gene rearrangements. Interestingly, mutations leading to stop codons in in-frame V gene rearrangements were detected in four cases. Since GC B cells acquiring such crippling mutations are usually efficiently eliminated within the GC, the finding of those mutations indicates that H-RS cells are derived from precursors within the GC that escaped apoptosis by a transforming event. PMID- 9209648 TI - NPM/ALK fusion mRNA expression in Hodgkin and Reed-Sternberg cells is rare but does occur: results from single-cell cDNA analysis. AB - BACKGROUND: The translocation t(2;5)(p23;q35) leads to the fusion of the nucleophosmin gene (NPM) on chromosome 5q35 to the recently described receptor kinase ALK on 2p23. It is characteristic of a subgroup of CD30+ large-cell anaplastic non-Hodgkin's lymphoma (ALCL). Since some cases of Hodgkin's disease (HD) and ALCL share common features, a common pathogenesis has been proposed in a report of the expression of NPM/ALK fusion mRNA in 11/13 Hodgkin's lymphomas. PATIENTS AND METHODS: We approached this question by micro-manipulatory isolation of single Hodgkin and Reed-Sternberg (H-RS) cells and subsequent RT-PCR amplification of NPM/ALK fusion cDNA from these single cells. RESULTS: Specificity of cell selection was shown by the HD-specific pattern of EBV-gene expression in single H-RS cells. In 4 out of 7 cases, NPM/ALK fusion cDNA was detected in the RNA from whole lymph node tissue. In 2 out of 9 cases, NPM/ALK fusion sequences were amplified from single H-RS cells, albeit in a very low frequency (< 5%). CONCLUSIONS: These data indicate that NPM/ALK fusion transcripts do not play an early role in the pathogenesis of HD. Whether the rare expression of NPM/ALK is the result of clonal heterogeneity or an indication for clonal evolution and progression toward ALCL can only be answered by the repeated analysis of indicator cases during the course of the disease. PMID- 9209649 TI - The role of eosinophils in the pathobiology of Hodgkin's disease. AB - BACKGROUND: Even though the presence of a prominent tissue eosinophilia represents a common histopathologic feature of Hodgkin's disease (HD), eosinophils have been mainly regarded as 'innocent' bystanders recruited and activated during the cellular reaction typical of HD. To evaluate the putative role of eosinophils or eosinophil-derived cytokines on tumor-cell regulation in HD, we have analyzed these cells for the functional expression of surface ligands (L) of the tumor necrosis factor (TNF) superfamily, whose specific receptors are known to transduce proliferation signals at the surface of Hodgkin (H) and Reed Sternberg (RS) cells. MATERIALS AND METHODS: Eosinophils from peripheral blood of healthy donors and patients with HD, primary hypereosinophilic syndrome (HES), or secondary hypereosinophilia (HE), were purified by density gradient centrifugation and immunomagnetic depletion of residual granulocytes. RESULTS: By immunostaining and mRNA analysis, we were able to show that eosinophils from normal donors and patients with HD, HES, and HE express a number of receptors and ligands of the TNF superfamily, including CD40, CD40L, CD30L, CD95/Fas, CD95/FasL and 4-1BB. In addition, we provide evidence that cytokines regulating eosinophil proliferation and activation, i.e., interleukin (IL)-5, IL-3, and granulocyte macrophage colony-stimulating factor, are able to enhance the cellular density of several TNF superfamily ligands and/or receptors at the surface of cultured eosinophils. Finally, we have shown that native CD40L and CD30L at the surface of purified eosinophils are functionally active and able to transduce proliferative signals on CD40+ and CD30+ target cells, including cultured H-RS cells. CONCLUSIONS: Our data suggest that eosinophils may act as important elements in the pathology of HD by providing cellular ligands for TNF-superfamily receptors (CD40, CD30, CD95/Fas) able to transduce proliferation and antiapoptotic signals at the surface of H-RS cells. The presence on eosinophils of receptors for TNF ligands expressed by activated T cells (i.e., OX40L, FasL, CD40L, 4-1BBL), also suggest that eosinophils may contribute to the deregulated network of interactive signals between H-RS cells, T cells, and other surrounding reactive cells. PMID- 9209650 TI - CD30-ligand and CD40-ligand expression in lymph nodes involved with Hodgkin's disease. AB - BACKGROUND: Reed-Sternberg cells of Hodgkin's disease express CD30 and CD40 receptors. The ligands for these receptors have been reported to have pleiotropic biologic activities in vitro, including induction of cell death and enhancing cell survival. Co-expression of the ligands for these receptors in lymph nodes involved with Hodgkin's disease is not known. PURPOSE: The purpose of this study was to examine CD30 ligand (L) and CD40L expression in lymph nodes of patients with Hodgkin's disease, and to study CD30L expression on nodal lymphocyte subsets. MATERIALS AND METHODS: CD30L expression on subsets of lymphocytes of five lymph nodes involved with Hodgkin's disease was determined by two-color FACScan. Messenger RNA expression of CD30L and CD40L was determined by the reverse-transcriptase polymerase chain reaction (RT-PCR) method performed on seven specimens involved with Hodgkin's disease (five lymph nodes and two spleens). RESULTS: Four of seven specimens (57%) contained cells that expressed CD30L mRNA and three specimens (43%) contained CD40L-expressing cells. The mean percentage of nodal lymphocytes expressing CD30L surface protein was < or = 20%. CONCLUSION: Hodgkin's disease lymph nodes and spleens frequently lack CD30L- and CD40L-expressing cells, and when CD30L is expressed, it is usually detected on few numbers of lymphocytes. The balance in the level of expression of these ligands in Hodgkin's disease lymph nodes may be related to the disease's clinical behavior. PMID- 9209651 TI - Bcl-6 protein expression in normal and neoplastic lymphoid tissues. AB - The human bcl-6 gene, which is rearranged in about 30% of diffuse large B-cell lymphomas (DLCL-B), encodes for a Kruppel-type zinc finger protein of 706 amino acids. In order to investigate the expression of the bcl-6 gene at the protein level, two monoclonal antibodies (PG-B6a and PG-B6p) directed against the human bcl-6 protein were generated by immunizing Balb/c mice with a recombinant protein corresponding to the amino-terminal region (amino acids 3-484) of bcl-6. PG-B6a (a = avian) recognized the most conserved bcl-6 epitope (expressed in many animal species, including avian). PG-B6p (p = paraffin) reacted with an epitope of bcl-6 partially resistant to fixatives and detectable on microwave-heated paraffin sections. At immunocytochemistry, bcl-6 localized in the nucleus with a microgranular or diffuse pattern. Strong nuclear expression of bcl-6 was mainly detected in normal germinal-center B-cells, whereas mantle- and marginal-zone B cells, as well as plasma cells and marrow B-cell precursors, did not express bcl 6. These immunohistological findings strongly suggest that bcl-6 may play a role as a regulator of germinal-center related functions. All MoAbs stained neoplastic cells of follicular lymphomas, DLCL-B, and Burkitt's lymphomas. In DLCL-B, bcl-6 expression was independent of bcl-6 gene rearrangements and did not correlate with expression of other markers or the proliferation index. Among low-grade B cell lymphomas, immunostaining for bcl-6 proved useful for differentiating proliferation centers in B-CLL (bcl-2+/bcl-6-) from trapped germinal centers in mantle-cell lymphomas (bcl-2-/bcl-6+). Strong nuclear positivity for bcl-6 was consistently detected in tumor (L&H) cells of nodular, lymphocyte-predominant Hodgkin's disease (NLPHD). These results further support the concept that NLPHD is a histogenetically distinct (germinal-center derived) subtype of HD. Notably, the nuclei of reactive CD3+/ CD4+ T cells near to and rosetting around L&H cells in NLPHD were also strongly bcl-6+, but lacked CD40 ligand (CD40L) expression. This staining pattern clearly differed from that of classic HD, whose cellular background was made up of CD3+/CD4+ T cells showing the bcl-6-/CD40L+ phenotype. The above immunohistological findings suggest that (a) bcl-6 may play a role in regulating B-cell differentiation step(s) occurring within germinal centers; (b) deregulated bcl-6 expression caused by rearrangements may contribute to B lymphomagenesis; (c) bcl-6 is possibly involved in the pathogenesis of NLPHD. PMID- 9209652 TI - Involvement of the bcl-6 gene in AIDS-related lymphomas. AB - BACKGROUND: Non-Hodgkin's lymphoma (NHL) represents a major complication of AIDS. Systemic AIDS-related NHLs (AIDS-NHLs) derive from B cells and are classified into four distinct groups, including small noncleaved-cell lymphoma (SNCCL), diffuse large-cell lymphoma (DLCL), anaplastic large-cell lymphoma (ALCL), and body-cavity-based lymphoma (BCBL). The molecular pathogenesis of AIDS-NHL is characterized by the association of specific genetic lesions with distinct AIDS NHL categories. Genetic lesions of AIDS-NHL involve proto-oncogenes (c-myc, Ras), tumor suppressor loci (p53, 6q), and viral infection (Epstein-Barr virus, human herpesvirus type 8). DESIGN: The aim of this work was to define the involvement of the bcl-6 gene in AIDS-related lymphomagenesis by investigating the distribution of bcl-6 structural alterations throughout the pathologic spectrum of AIDS-NHL. Both gross rearrangements and mutations in the 5' noncoding regions of the gene were investigated. RESULTS: Gross rearrangements of bcl-6 are confined to a fraction of AIDS-DLCL cases among AIDS-NHLs. Conversely, mutations of the 5' noncoding regions of bcl-6 are detected in a large proportion of AIDS SNCCLs, AIDS-DLCLs and AIDS-ALCLs independent of the concomitant presence of bcl 6 rearrangements. CONCLUSIONS: Mutations of the 5' noncoding regions of bcl-6 represent the most frequent genetic lesion presently detectable among systemic AIDS-NHLs. The frequency of these mutations and their location in the proximity of bcl-6 regulatory regions suggest that they may play a role in AIDS-related lymphomagenesis. PMID- 9209653 TI - Bcl-1/cyclin D1 in malignant lymphoma. AB - Mantle-cell lymphoma comprises 2%-10% of all non-Hodgkin's lymphomas (NHLs). Patients present with generalized disease, and have a poor prognosis. Three different histologic patterns (mantle zone, nodular, and diffuse) and three different cytological variants (classical, blastic, and pleomorphic) have been described. The phenotype (strong surface IgM, CD5+, CD10-, CD23-, cyclin D1+ and B-cell markers+) is remarkably constant. Dependent on the methods used (PCR, Southern blot analysis, and cytogenetics) a t(11;14) can be detected in approximately 35%-66% of cases. Using FISH analysis, possibly almost all cyclin D1-expressing MCLs carry this translocation, indicating that a substantial part of these translocations are missed by conventional methods. This has been confirmed by DNA fiber FISH analysis by which the breakpoints could be accurately mapped over a 220 kb region centromeric of the cyclin D1 gene. Additional genetic abnormalities involve breakpoints and deletion at the 3' end of the cyclin D1 gene, numerical chromosomal aberrations, mutations in p53, and deletions of p16. These may be associated with tumor progression. Owing to the translocation t(11;14), the cyclin D1 gene is activated. At the RNA level, approximately 90% of MCLs show overexpression. This corroborates immunohistochemistry on paraffin tissue sections. Since expression of cyclin D1 in normal lymphoid cells is very low to undetectable, and only hairy-cell leukemia and very few other B-cell lymphomas show expression, immunohistochemistry for cyclin D1 provides an excellent marker for MCL. In hairy-cell leukemia, expression is moderate and cannot be explained by chromosomal translocation. PMID- 9209654 TI - Somatic mutations of the translocated bcl-2 gene are associated with morphologic transformation of follicular lymphoma to diffuse large-cell lymphoma. AB - BACKGROUND: Ninety percent of low-grade follicular lymphomas (FLs) carry the t(14;18) translocation. This event juxtaposes the bcl-2 oncogene to the immunoglobulin (Ig) heavy-chain gene and leads to bcl-2 gene overexpression. Morphologic transformation of FL to high-grade lymphoma is associated with multiple secondary chromosomal abnormalities of the neoplastic cells. DESIGN: To analyze whether additional structural alterations of the translocated bcl-2 gene are associated with morphologic transformation of FL, we PCR-amplified, cloned, and sequenced the major breakpoint region (MBR) and the open reading frames (ORF) of the translocated bcl-2 oncogene in six paired samples of FL and subsequent diffuse large-cell lymphoma (DLL). RESULTS: In five cases, FL and DLL cells were clonally related, as suggested by the identical MBR sequences, but in one case they were different. PCR single-strand conformation polymorphism (SSCP) and sequence analyses were performed for identification of structural alterations of the bcl-2 gene in the OFR region corresponding to the 239 amino acid p26-bcl-2a protein. In three of the six patients, a total of 11 point mutations of the ORF were detected in the DLL cells. Four of them, at positions 29, 46, 59, and 106, yielded amino acid replacements. CONCLUSIONS: These findings demonstrate that FL and DLL cells may be clonally related or unrelated. They also show that transformation of FL cells can be associated with somatic point mutations of the bcl-2 oncogene ORF sequence resulting in alteration of the p26-bcl-2a gene product. PMID- 9209655 TI - The Kaposi's sarcoma-associated herpesvirus (human herpesvirus-8) in Kaposi's sarcoma, malignant lymphoma, and other diseases. AB - BACKGROUND: Two novel nonhuman DNA fragments were discovered in an AIDS-related Kaposi's sarcoma (KS) lesion using representational difference analysis. DESIGN: These sequences belong to a previously unidentified gamma-2-herpesvirus exhibiting homology with Herpesvirus saimiri and Epstein-Barr virus. This virus was named Kaposi's sarcoma-associated herpesvirus (KSHV) and provisionally designated human herpesvirus-8 (HHV-8). RESULTS: KSHV is detectable in more than 90% of classical-Mediterranean, iatrogenic, endemic-African, and AIDS-epidemic KS lesions. In situ PCR studies have demonstrated KSHV in the spindle cells and endothelial cells of KS lesions. KSHV appears to be a transmissible B lymphotropic herpesvirus. It is detectable in circulating B cells in some HIV infected patients, and this finding appears to predict the future development of KS among these individuals. KSHV has been identified in a rare and distinct subset of malignant lymphoma referred to as body cavity-based lymphoma but not in other lymphoid neoplasms. KSHV is absent from most other HIV- and non-HIV associated lymphadenopathies. CONCLUSIONS: Further studies should lead to a better understanding of the role of KSHV in the pathogenesis of these disorders and may eventually show that KSHV represents the long sought-after etiologic agent of Kaposi's sarcoma. PMID- 9209656 TI - Integration of Epstein-Barr virus in Burkitt's lymphoma cells leads to a region of enhanced chromosome instability. AB - BACKGROUND: Several lymphomas are associated with Epstein-Barr virus (EBV) infection. However EBV is not detectable in 100% of cases using standard staining techniques. It still remains an open question whether in these EBV-negative cases EBV has never infected the cell, whether it has infected the cell and escapes conventional screening methods, or whether it has been lost again after initial infection. MATERIALS AND METHODS: The physical status of EBV in the Burkitt's lymphoma cell line BL60-P7 as well as in three somatic cell hybrids between BL60 P7 and its autologous EBV-immortalized lymphoblastoid cell line IARC 277 was analyzed using conventional cytogenetics, Southern blotting, and fluorescence in situ hybridization (FISH). RESULTS: Integration of EBV into the host genome of the lymphoma cell line BL60-P7 leads to an achromatic gap which causes a 'vulnerable site'. In hybrid cells, loss of integrated EBV, together with an adjacent chromosomal fragment, occurs during long-term cultivation. The integrated EBV genome, including genes encoding for LMP and EBER, is partly deleted. CONCLUSION: We assume that integration of EBV into the host cell genome could be a more common event in lymphoma cells. Partially deleted EBV might escape standard detection assays. The integration might constitute a chromosomal region prone to break events akin to the phenomenon of fragile sites, leading to the loss of viral DNA as well as chromosomal DNA. This observation makes it tempting to speculate that under certain conditions EBV can act in lymphomagenesis by a so-called hit-and-run mechanism. PMID- 9209657 TI - Characteristics of human EBV-specific cytotoxic T lymphocytes utilized for adoptive immunotherapy of EBV-induced lymphoproliferations in xenografted SCID mice. AB - Human Epstein-Barr virus-specific cytotoxic T lymphocytes (EBV-CTLs) prolong the survival of mice with severe combined immune deficiency bearing the autologous, but not HLA-mismatched, human EBV-induced lymphoproliferative disorders (EBV LPDs). In the present study, we demonstrate that the HLA-restricted activity displayed by EBV-CTLs both in vitro and in vivo correlates with their in vivo homing pattern, and further characterize these effectors. EBV-CTLs were CD3+, CD16/56-, TCR alpha/beta+, predominantly CD8+ and CD4-, and had a high expression of T-cell activation antigens. EBV-CTLs were positive for CD11a/CD18, CD54, CD58, CD44, CD49d, CD28, and CD45RO, and negative for CD45RA, CD11b, CD11c. After 26 days in culture, EBV-CTLs displayed strong cytotoxicity against the autologous EBV-transformed B-cell line (EBV-LCL), which was inhibited by the addition of anti-CD3 MoAb and mostly HLA class I-restricted. Unirradiated and irradiated EBV CTLs in the absence of IL-2 failed to proliferate after more than 2 days in culture with the autologous EBV-LCLs, while unirradiated EBV-CTLs with IL-2 formed large colonies and had a high thymidine incorporation both on days 5 and 8. The cytotoxicity of irradiated EBV-CTLs against the autologous EBV-LCLs was conserved. It remains to be determined whether irradiated EBV-CTLs are capable of homing to EBV-LPDs in vivo and to mediate a therapeutic response comparable to that observed with unirradiated EBV-CTLs. PMID- 9209658 TI - How many more nails to seal the coffin of dose intensity? PMID- 9209659 TI - Drug resistance markers: are they bad or good? PMID- 9209660 TI - Elective surgery for gastrointestinal tumours in the elderly. AB - The geriatric population is expanding and clinical decision-making is often complicated by the effects of ageing. Age should not be the only parameter considered when addressing medical problems. Elderly subjects have been denied surgery because of their presumed higher mortality and morbidity. The present review summarises the physiology of the aged and discusses operative risks, mortality and morbidity rates as well as therapeutic results for the different gastrointestinal sites when affected by cancer. Reports on surgical treatments are revisited and compared to the same procedures delivered to younger patients in the context of the ethical issue of offering the best care to every patient. Elective operations by surgical oncologists are found to be safe with the exception of major liver resections. Complication rates and mean hospital stay do not differ between the two age groups provided the procedure is conducted with the best-known technique in expert hands. A drop in operative morbidity has occurred in the past three decades. Several investigators have emphasised the marked increase in morbidity and mortality experienced by elderly patients when undergoing emergency procedures. Associated diseases have to be properly assessed, as the elderly have a frail physiological balance with a reduced capacity for recovery from traumatic events including major surgical procedures. Careful preoperative evaluation, intraoperative conduct and postoperative care are presently achieved in almost every major hospital. Good clinical practice is based on the balance between probability of cure and toxic effects. Treatment of the elderly should no longer be based on untested beliefs and personal opinions. The elderly should be accrued for prospective clinical evaluation and should not be denied optimal surgical treatment. PMID- 9209661 TI - A randomized trial of five versus eight courses of cisplatin or carboplatin in advanced epithelial ovarian carcinoma. A North Thames Ovary Group Study. AB - BACKGROUND: There is no clear data on the optimum number of courses of platinum chemotherapy required for treating advanced ovarian cancer. A prospective randomised trial was performed to compare five with eight courses of either carboplatin 400 mg/m2 or cisplatin 75 mg/m2 given four-weekly to patients with stage IC-IV ovarian cancer. PATIENTS AND METHODS: 225 patients were entered into the study and 233 were eligible for randomisation: 118 to five courses and 115 to eight courses. In each randomisation, half the patients received carboplatin and half received cisplatin. RESULTS: The mean number of courses received on the five arm was 4.74 and on the eight arm, 6.42, an increase of 35%. The median survival for all patients was 24 months with the median survival for the 156 patients with stage 3 disease being 21 months. No difference was detected in survival (P = 0.53) or time to progression from initial surgery (P = 0.29) between the two arms of the trial. False-negative calculations based on a multivariate analysis show that the trial currently has 95% power for excluding a difference of 10% in favour of the eight course arm at three years. CONCLUSION: There is insufficient evidence at the present time to justify more than five courses of first-line single agent platinum chemotherapy in the management of advanced ovarian cancer. PMID- 9209662 TI - Breast cancer: pretreatment drug resistance parameters (GSH-system, ATase, P glycoprotein) in tumor tissue and their correlation with clinical and prognostic characteristics. AB - BACKGROUND: The identification of new factors predicting relapse, outcome and response to systemic therapy in breast cancer is warranted. The measurement of biological markers such as drug resistance parameters (DRPs), which are part of the phenotype of malignant cells and contribute to resistance to anti-cancer drugs may be a possibility, which may ultimately lead to improvement of therapeutic results. PATIENTS AND METHODS: The level of glutathione (GSH), activities of glutathione-S-transferase (GST), glutathione-peroxidase (GPx), 06 alkylguanine-DNA-alkyltransferase (ATase), and P-glycoprotein (PGP) were measured in tumor and adjacent tumor free tissue samples from 89 consecutive, untreated females with breast cancer and correlated with clinical and prognostic factors. Early breast cancer (EBC) was diagnosed in 56 patients, 22 patients had locally advanced (LABC) and 11 patients metastatic breast cancer. RESULTS: All DRPs showed significantly higher expression in tumor than in tumor free tissues. GPx was positively correlated with GST (r = 0.3, P = 0.0048) and with GSH (r = 0.5, P = 0.0001) in tumor as well as in normal tissue. GST activity was significantly higher in EBC than in LABC or metastatic breast cancer (P = 0.02). GSH level was significantly higher in grade 1 than in grade 2 or grade 3 tumors (P = 0.01). When clinical characteristics were related to the level of DRP, 'high' GSH was associated with age > 60 years (P = 0.01) in EBC, and with grade 1-2 tumors (P = 0.05) in LABC. No differences in OS were apparent between groups of 'high' and 'low' DRP-expression. However, the four-year estimated disease-free survival of EBC tended to be higher in patients with 'high' GST (P = 0.10) and of LABC in patients with 'high' GPx levels (P = 0.06). CONCLUSION: We conclude that 'high' levels of DRP in tumor tissue of breast cancer patients are part of the initial phenotype of the malignant cells. Due to its high prevalence (83% in EBC, 100% in primarily metastatic breast cancer), PGP did not add to prognostic information. High levels of GSH, GST and GPx were associated with favorable clinical characteristics and good prognosis, whereas low levels of GSH and GST activity were associated with more aggressive or more advanced disease. PMID- 9209664 TI - Phase II study of the combination carboplatin and paclitaxel in patients with ovarian cancer. AB - BACKGROUND: Recently the feasibility of combining carboplatin with paclitaxel has been demonstrated in dose-finding studies. Maximum tolerated doses were 550 mg/m2 and 200 mg/m2 (three hours), respectively. We report now a phase II study in ovarian cancer patients. PATIENTS AND METHODS: Twenty-one chemo-naive patients with optimally (n = 6) or suboptimally (n = 15) debulked stage III or IV ovarian cancer were treated every three weeks for six courses with paclitaxel (200 mg/m2) as a three-hour infusion, immediately followed by carboplatin (550 mg/m2) as a 30 minute infusion. RESULTS: Uncomplicated neutropenia was the principal toxicity, with mild anemia occurring regularly. As observed in the preceding phase I study, a relative lack of thrombocytopenia, generally grade III was found. Other toxicities consisted of mild neurotoxicity, nausea and vomiting, alopecia, myalgia, and bone pain. All suboptimally debulked patients responded to therapy. Overall, 12 patients underwent second-look laparoscopy, which revealed a pathologically confirmed complete remission in six. The median follow-up interval at the time of analysis was 14 months. Twelve patients are currently free of progression, at 8+ to 19 +/- months after the start of therapy. CONCLUSION: The carboplatin/paclitaxel combination appears to be a well-tolerated regimen, yielding high response rates. This combination has now gone forward to be evaluated in prospective randomized trials versus the cisplatin/paclitaxel combination. PMID- 9209665 TI - Phase I/II study of the combination of carboplatin and paclitaxel as first-line chemotherapy in patients with advanced epithelial ovarian cancer. AB - PURPOSE: We performed a phase I/II study evaluating the combination of paclitaxel and carboplatin as first-line chemotherapy in patients with advanced ovarian cancer. The aim of this study was to define a feasible and safe combination regimen that could be recommended for future phase III studies. DESIGN: This study was a parallel two-arm, non-randomized, open trial. In a first step, carboplatin was administered at a fixed dose of AUC 5 and paclitaxel was escalated in 25 mg/ m2 steps starting at 135 mg/m2. Paclitaxel was given as a three-hour infusion. Carboplatin was administered on day 1 following paclitaxel in one study arm and 24 hours after paclitaxel infusion on day 2 in the other study arm. Carboplatin was escalated to AUC 6 and AUC 7.5 after the MTD for paclitaxel had been defined. Treatment was repeated every three weeks. PATIENTS: Sixty-one patients with untreated histologically confirmed epithelial ovarian cancer were recruited of whom 59 were found eligible and evaluable for toxicity. Thirty-three patients with bidimensionally measurable disease were evaluable for tumor response. RESULTS: We could not detect any advantage of the two-day schedule compared with the more convenient one-day schedule. Dose limiting toxicities were neutropenia, thrombocytopenia, and neurotoxicity. Except for two patients, toxicity was acceptable and clinically managable. One patient died of neutropenic sepsis and one further patient developed grade III peripheral neurotoxicity that did not resolve within two months after chemotherapy had been terminated. Overall objective response rate was 70%. The MTD for paclitaxel was 185 mg/m2 and AUC 6 for carboplatin, respectively. Secondary prophylaxis with G CSF did not allow further dose escalation and therefore is not generally recommended. CONCLUSIONS: Paclitaxel 185 mg/m2 given as three-hour infusion followed by carboplatin AUC 6 is a feasible and safe regimen and can be recommended for phase III trials. Observed response rates justify further evaluation of this combination. A randomized phase III trial comparing a three hour infusion of paclitaxel 185 mg/m2 combined with either carboplatin AUC 6 or cisplatin 75 mg/m2 as first-line chemotherapy of advanced ovarian cancer has recently been initiated by our group. PMID- 9209663 TI - Glutathione S-transferase activity in epithelial ovarian cancer: association with response to chemotherapy and disease outcome. AB - BACKGROUND: Conflicting data have been reported about the association between glutathione S-transferase (GST), a family of proteins implicated in detoxification of cytotoxic drugs in human ovarian in vitro models, and response to chemotherapy and prognosis in ovarian cancer patients. The aim of this study was to analyze the possible clinical role of GST activity in a large series of primary ovarian cancer patients. PATIENTS AND METHODS: The study included a large series of primary untreated ovarian cancer patients who underwent cytoreductive surgery and chemotherapy and who were followed up in a single institution. GST activity levels were assessed in tumor extracts by using a biochemical assay. A cut-off of 250 units of enzymatic activity was chosen according to the receiver operating characteristics (ROC) curve. RESULTS: GST activity levels were distributed in an asymmetrical manner (median: 266 units; range: 4-918 units) and did not seem to be associated with stage, histopathological grading, ascites, or residual tumor after surgery. Higher GST activity levels were found in patients who responded to chemotherapy (median: 298 units, range: 50-691) than in those who responded only partially (median: 227 units, range: 19-747) or not at all to chemotherapy (median: 246 units, range: 4-811) (H = 7.02, P = 0.029). Moreover, the percentage of cases with > 250 units was significantly higher among complete responders (66%) than partial responders (37%) or non-responders (48%) (chi 2 = 7.32; P = 0.025). When multivariate analysis, including clinico-pathological parameters and GST activity status as predictors of response to chemotherapy, was carried out, residual tumor, stage and GST status retained independent predictive value. Patients with high GST activity had more favourable prognosis than those with low GST activity. The median PFS was 42 months for patients with high GST activity compared to 17 months for those with low GST activity (P = 0.037). The median overall survival was 72 months for high-GST-activity and 42 months for low GST-activity patients (P = 0.043). Substantially similar results were obtained in the subgroup of stage II-III-IV ovarian cancer patients. Multivariate analysis including the clinico-pathological parameters and GST activity status was performed in stage III-IV ovarian cancer patients: Stage IV disease, residual tumor > 2 cm, the presence of ascites and low GST activity status retained independent negative prognostic roles. CONCLUSION: A direct association between high GST activity and a better clinical outcome in terms of response to chemotherapy and survival has been observed in a large series of primary untreated ovarian cancer patients. These results, which are contrary to the expectations raised by in vitro studies, emphasize the need for caution when translating in vitro-generated hypotheses to the clinical setting. PMID- 9209666 TI - Phase II trial of tirapazamine combined with cisplatin in chemotherapy of advanced malignant melanoma. AB - PURPOSE: A phase II study was undertaken to determine the efficacy of tirapazamine (TPZ) combined with cisplatin (cDDP) in patients with metastatic melanoma. PATIENTS AND METHODS: Between June 1994 and November 1995, 48 patients with metastatic melanoma were treated with TPZ (260 mg/m2, administered intravenously over two hours) followed in one-hour by cDDP (75 mg/m2 over one hour) every 21 days. Sixteen patients had received prior chemotherapy, and 13 of these had failed to respond to prior cDDP. None of the patients had symptomatic brain metastasis. RESULTS: Nine patients had partial responses, with an overall response rate of 19% (95% confidence interval (95% CI) of 9%-33%). The median duration of response was six months. None of the responders had received prior chemotherapy. Responses were seen in 8 (33%, confidence interval of 16%-55%) of 24 patients with primary cutaneous melanoma who had received no prior chemotherapy and in the only patient with previously untreated conjunctival melanoma. There were no responders among the seven patients with choroidal melanoma and 16 patients with previously treated cutaneous melanoma. Two patients with partial responses were rendered free of gross disease surgically three months after completing eight courses of TPZ-cDDP; they remain free of tumor recurrence. Responses were seen in lymph nodes (27%), lung (26%), skin (20%), adrenal gland (20%), soft tissues (17%) and liver (17%). Common toxicities included muscle cramps, fatigue, gastrointestinal effects and peripheral neuropathy. Fatigue, nausea, vomiting, anorexia, and muscle cramps were grade 3 or 4 in less than 10% of the courses. Neutropenia and thrombocytopenia were rare. CONCLUSION: The TPZ-cDDP combination has definite activity against chemotherapy naive patients with cutaneous melanoma and warrant further studies in combination with other cytotoxic agents. PMID- 9209667 TI - Expression of MAGE and BAGE genes in Japanese breast cancers. AB - BACKGROUND: The MAGE and BAGE genes code for distinct antigens, which are recognized on melanoma cells as well as on other various tumor cells by autologous cytolytic T lymphocytes. These antigens may thus constitute useful targets for specific immunotherapy, since no expression of MAGE or BAGE genes has been recognized in normal tissue except for the testis. PATIENTS AND METHODS: We studied the MAGE-1, MAGE-3, and BAGE gene expression observed in 49 Japanese breast cancers. Gene expression was evaluated by reverse transcription polymerase chain reaction. RESULTS: Out of 49 tumor tissue specimens of primary breast cancers, the expression of MAGE-1, -3 and BAGE was recognized in 15 (31%), 12 (24%), and 4 (8%) tumors, respectively. The expression of MAGE and BAGE genes is not recognized in normal breast tissue. The expression of the MAGE-3 gene was frequently recognized in tumors with lymphatic and/or vascular vessel permeations. Either MAGE-1 or -3 gene expressions were induced in 1 of 3 MAGE-1 negative breast cell lines or 1 of 3 MAGE-3 negative breast cell lines by the treatment with 5-aza-2'-deoxycytidine. CONCLUSIONS: These findings suggest that: 1) the identification of such antigens coded by MAGE or BAGE genes may thus offer the possibility of using specific immunotherapy, and 2) the use of a demethylating agent may increase the number of patients who might be candidates for MAGE specific immunotherapy. PMID- 9209668 TI - 5-Fluorouracil, interferon-alpha-2b and cisplatin (FAP) for advanced urothelial cancer. A phase II study. Hellenic Co-operative Oncology Group. AB - PURPOSE: To evaluate the efficacy and toxicity of the FAP combination chemotherapy as first-line treatment in advanced urothelial cancer. PATIENTS AND METHODS: Thirty-four patients with histologically confirmed advanced urothelial cancer, with measurable disease and without previous chemotherapy entered the study; all 34 are evaluable. The 28 males and 6 females had a median age of 65 (19-75) and a median ECOG performance status of 1 (0-2). Twenty-eight patients had bladder cancer, four had renal pelvic cancer and two ureteral cancer. Thirty patients had transitional cell carcinoma and four mixed, mostly of grade 3. Sites of disease included lymph nodes (18), bladder (9), liver (9), pelvic mass (9), lung (7), etc. The treatment plan was as follows: 5-fluorouracil 500 mg/m2 continuous infusion D1-D5 and D22-D26; interferon-alpha-2b 5 million i.u./m2 D1 D5 followed by 3x/week and then D22-D26; cisplatin 25 mg/m2 D1, D8, D15, D22. Cycles were repeated every 36 days. RESULTS: The median number of cycles administered was 3 (1-6). The relative dose intensities for 5-fluorouracil, interferon and cisplatin were 76%, 71% and 75%, respectively. Twenty-two of 34 patients (65%, 95% confidence interval [95% CI], 46% to 80%) had objective responses, including six complete clinical responses (CR) (18%, 95% CI, 7% to 35%) and 16 partial responses (PR) (47%, 95% CI, 30% to 65%). Three patients had stable disease and seven progressed. Two patients discontinued treatment after the first cycle because of toxicity. The median survival is 15.30 months (1.40 37.60), the median time to progression 11.60 months (4.13-37.60), and the median survival of complete responders 20.75+ months (8+ to 38+). The only significant hematologic toxicity was the grade 3-4 neutropenia in 44%. Non-hematologic toxic effects were unremarkable. CONCLUSION: The FAP combination as first-line chemotherapy is highly active in the treatment of advanced urothelial cancer, and has limited toxicity. Further phase III studies are in progress to compare FAP and M-VAC. PMID- 9209669 TI - Fludarabine-mitoxantrone combination-containing regimen in recurrent low-grade non-Hodgkin's lymphoma. AB - BACKGROUND: The promising results of fludarabine (FLU) in chronic lymphocytic leukemia have prompted its extensive evaluation in low-grade non-Hodgkin's lymphoma (LG-NHL). Its different mechanisms of action make FLU an attractive partner for combination with other cytostatic agents. PATIENTS AND METHODS: We used a three-drug combination of FLU (25 mg/m2 i.v. on days one to three), mitoxantrone (10 mg/m2 i.v. on day one) and prednisone (40 mg given orally on days one to five) (FMP) to treat 48 patients with recurrent LG-NHL. RESULTS: Of the 48 patients, 17 (35%) achieved complete responses (CR), 23 (48%) partial responses, while the remaining 8 (17%) showed no benefit from the treatment. The risk of lower CR rate was significantly correlated with the presence of advanced stage (IV) (P = 0.01), the number of previous regimens (> or = 3) (P = 0.006), and the follicular histologic subtype (P = 0.02). The major toxic effects observed were neutropenia and infections; there was only one fatality, due to drug-related side effects. CONCLUSIONS: These data confirm the significant efficacy of the FMP fludarabine-mitoxantrone combination regimen in obtaining a good remission rate with moderate toxicity in a particular subset of recurrent LG NHL. PMID- 9209670 TI - Carboplatin and cisplatin pharmacokinetics after intrapleural combination treatment in patients with malignant pleural effusion. AB - BACKGROUND: Cisplatin (DDP) and carboplatin (CBDCA) are two of the most effective drugs in a locoregional approach. Since simultaneous combined treatment with intrapleural DDP and CBDCA has not been reported in humans, we investigated its use in patients with malignant effusions, focusing on pharmacokinetics. PATIENTS AND METHODS: The pharmacokinetics of DDP and CBDCA were studied in 10 patients with malignant pleural effusion treated intrapleurally with a combination of DDP (60 mg/m2) and CBDCA (270 mg/m2) and in additional patients who received the same doses of drugs administered intravenously as single agents or in combination. Platinum (Pt) species originating from DDP (metabolites plus unchanged DDP) and intact CBDCA in plasma and pleural fluid ultrafiltrates were measured by means of high performance liquid chromatography and atomic absorption spectrometry. RESULTS: Both in the plasma and pleural fluid, the total levels of free Pt represented the additive result of the individual concentrations of CBDCA and Pt species derived from DDP. After intrapleural combination, high pleural-plasma ratios of the peak concentrations and AUCs were observed both for CBDCA and DDP derived Pt species, highlighting a distinct local pharmacological advantage. However, the Pt species originating from DDP were absorbed more rapidly from the pleural cavity than CBDCA (Ka = 86 x 10(-3) vs. 37 x 10(-3) min-1, P < 0.05). Intrapleural combination of CBDCA and DDP produced therapeutic plasma levels of reactive (free) DDP species and increased the extent of their residence time (MRT) compared with single intravenous DDP treatment [peak concentration: 1.1 +/- 0.1 (SD) vs. 1.6 +/- 0.2 microgram/ml; MRT: 5.2 +/- 1.9 vs. 0.5 +/- 0.06 h]. Furthermore, the plasma AUC of free CBDCA after intrapleural combined treatment (2.1 +/- 0.5 mg/ ml x min) was similar to that after intravenous administration of CBDCA alone (2.1 +/- 0.2 mg/ml x min). The intrapleural treatment was well tolerated by all patients. Toxicity consisted of mild nausea and vomiting (grade 1-2 according to the WHO scale) in four patients. Myelosuppression (grade 1-2) was remarkable only in two heavily pretreated patients. No evidence of recurrence of the pleural effusion was observed in six patients (complete response), while an asymptomatic minimal fluid reaccumulation not requiring drainage (partial response) was observed in four patients. CONCLUSIONS: The pharmacologic results seem to exclude a pharmacokinetic interaction between CBDCA and DDP and suggest that a dose of CBDCA 2-fold higher than that used in this study associated intrapleurally with 60 mg/m2 DDP could induce an acceptable and predictable myelosuppression. PMID- 9209671 TI - Liver metastases from colorectal cancer. PMID- 9209672 TI - Sequential high-dose therapy and autologous stem cell transplantation for treatment of mantle cell lymphoma. AB - BACKGROUND: Mantle cell lymphoma (MC) is not curable with conventional chemotherapy. To improve the prognosis of patients with this disease, we prospectively studied an intensive sequential therapy consisting of the Dexa-BEAM regimen (dexamethasone, BCNU, etoposide, ara-C, melphalan) followed by myeloablative therapy with autologous stem cell reinfusion. PATIENTS AND METHODS: Nine consecutive patients with stage III/IV MC were included. Two had untreated disease, four were in first remission, whereas three had more advanced disease. All patients underwent one to two cycles of Dexa-BEAM chemotherapy to reduce the tumor load and to mobilize peripheral blood progenitor cells (PBPC). Subsequently, patients were treated with high-dose radiochemotherapy followed by PBPC reinfusion and were prospectively analyzed for residual disease by clinical methods as well as by PCR amplification clonal CDRIII rearrangements. RESULTS: With an overall response rate of 100%, the initial Dexa-BEAM cycles effectively reduced the tumor load. All patients proceeded to high-dose therapy and subsequent stem cell rescue. Engraftment was prompt, and procedure-related deaths did not occur. With a median follow-up of 12 (3-33) months post transplant, all patients are alive in continuing clinical and molecular remission. CONCLUSIONS: Sequential intensive therapy consisting of Dexa-BEAM and high-dose radiochemotherapy appears to be a highly effective treatment for patients with MC. However, the data are still preliminary, and larger patient numbers and a longer follow-up are required. PMID- 9209673 TI - Citrobacter freundii and fatal neutropenic enterocolitis following adjuvant chemotherapy for breast cancer. PMID- 9209674 TI - Interferon-alpha-2b in the management of patients with relapsed and/or refractory Hodgkin's disease. PMID- 9209675 TI - Identification of a novel keratinocyte mitogen derived from bovine pituitary glands. AB - Bovine pituitary glands contain one or more factors that are important for keratinocyte proliferation in serum-free culture medium. We used a tissue culture system in which the growth of keratinocytes in basal medium (KBM) containing insulin was dependent upon exogenous growth factors. Using this experimental system, we began to purify and characterize the pituitary factor(s) necessary for clonal growth of human keratinocytes in serum-free medium. Proteins of approximately 150 kDa and 95 kDa bound specifically to living keratinocytes, and we suggest that the 95 kDa protein is a likely novel mitogen. Although prolactin has been previously identified as a pituitary hormone that may act as an in vitro mitogen for keratinocytes, imunoblots indicated that the 95 kDa protein was unrelated to prolactin. Furthermore, the 95 kDa protein showed high homology with a bovine 90 kDa heat shock protein in the limited sequencing of an internal peptide. PMID- 9209676 TI - Epidermal growth factor and temperature regulate keratinocyte differentiation. AB - The limited life-span and irregularities in epidermal differentiation and barrier function that have restricted the utility of presently available skin culture models for pharmacological and toxicological studies indicate that further modifications of culture conditions are required for optimization of these models. In the present study epidermis reconstructed on de-epidermized dermis was used to investigate the effects of temperature and epidermal growth factor (EGF) on epidermal differentiation and lipogenesis. When cultured at 37 degrees C, keratinocytes formed a well-differentiated epidermis whether EGF was present or not. However, the thickness of the epidermis, particularly of the stratum corneum, was higher in the presence of EGF. Both the differentiation-specific protein markers (keratins 1 and 10, involucrin and transglutaminase) and lipid markers (ceramides) were synthesized. EGF-induced increases in triglyceride content caused accumulation of lipid droplets within the stratum corneum which is indicative of a hyperproliferative effect of EGF. In the absence of EGF, a well differentiated epidermis was generated at 33 degrees C with a morphology showing a higher resemblance to native epidermis than cultures grown at 37 degrees C. The stratum corneum was less compact and with practically no lipid droplets, irregularly shaped keratohyalin granules were abundant in the stratum granulosum, lamellar body extrusion was improved and the number of stratum corneum layers was reduced to normal levels. However, EGF supplementation had a deleterious effect on epidermal morphogenesis and differentiation of cultures grown at 33 degrees C. The epidermis lacked a stratum granulosum and the stratum corneum contained a high number of nuclear remnants. The synthesis of the early specific protein differentiation markers (keratins 1 and 10) was suppressed on both the protein and mRNA levels without significant interference with the synthesis of late differentiation lipid markers, such as ceramides. From this observation it can be concluded that the synthesis of keratins associated with terminal differentiation is profoundly affected by the presence of EGF and is sensitive to temperature and that of ceramides is not. The finding that TGF alpha did not modulate the morphogenesis and synthesis of keratins 1 and 10 in cultures grown at 33 degrees C indicates possible differences between the postreceptor binding processes of these EGF receptor ligands. PMID- 9209677 TI - T-cell population of primary and secondary cutaneous B-cell lymphomas does not express the cutaneous lymphocyte-associated antigen (CLA). AB - Primary cutaneous B-cell lymphomas (CBCL) are a group of malignant lymphomas with apparently distinct clinicopathological and immunophenotypical features. As in other B-cell lymphomas, the accompanying benign cell population in CBCL includes a variable number of T lymphocytes whose role is not well understood. In the present study we characterized the immunophenotype of these T cells and compared it with that of the reactive T-cell population in specific skin involvement by noncutaneous B-cell malignancies. Our results indicated that most T cells in both primary and secondary B-cell lymphomas were CLA+ memory/effector helper T cells which differed from the currently known CLA+ memory/effector helper T lymphocytes of the skin-associated lymphoid tissue (SALT) system. However, the endothelial CLA ligand, E-selectin, was expressed on dermal vessels. These results suggest that a B cell environment and/or a lack of epidermal involvement promote(s) the recruitment into the skin of a different, apparently less specific, subset of memory helper T cells from those seen in T-cell-mediated dermatoses. PMID- 9209678 TI - Epicutaneous application of leukotriene B4 induces patterns of tenascin and a heparan sulfate proteoglycan epitope that are typical for psoriatic lesions. AB - Application of leukotriene B4 (LTB4) to normal human skin induces changes similar to those found in psoriatic skin, and it has proved to be a useful model for studying the pathogenesis and treatment of psoriasis. We studied the expression patterns of molecules that have recently been shown to be altered in lesional psoriatic skin, including the extracellular matrix protein tenascin (TN) and the basement membrane and cell surface-associated heparan sulfate proteoglycans (HSPGs). During 72-h the expression of these markers was studied immunohistochemically and the expression of TN was correlated with epidermal proliferation and influx of inflammatory cells. Following the peak influx of polymorphonuclear leukocytes, a marked increase in TN expression was noted in the papillary dermis 72 h after LTB4 application. The expression patterns of basal membrane-associated epitopes of HSPG remained unaltered, whereas the expression of cell surface-associated HSPG disappeared 72 h after LTB4 application. A significant correlation was found between dermal TN expression and epidermal hyperproliferation, and between TN expression and the presence of dermal T cells. These findings indicate that the model of LTB4-induced acute cutaneous inflammation displays many characteristics of early psoriatic lesions and could serve as a model to study some of the cell biological changes in this disease. PMID- 9209679 TI - Clinical features and age distribution of patients with HPV 2/27/57-induced common warts. AB - The morphology of common warts depends on the inducing human papillomavirus (HPV) type. In order to assess the impact of the virus type on wart epidemiology we determined the virus type by PCR and recorded anamnestic data of 238 patients with common warts. Warts induced by the related HPV types 2, 27 and 57 predominated in the study population (n = 202). These warts mostly occurred as multiple verrucae vulgares, mosaic warts or endophytic warts. Patients aged between 10 and 30 years were most affected and they typically displayed a long disease history (mean duration of warts at the time of first clinical examination, 22 months). A different age distribution was observed in HPV 1 induced warts, most of which occurred in children 6-10 years of age. HPV 2 related warts responded only modestly to treatment, as they persisted in approximately 50% of all patients for more than 6 additional months. No sex preference was detected, but an association with atopic diseases was noted as 39.8% of patients with warts containing HPV 2-related viruses showed a history of atopic eczema, pollinosis or asthma as compared with 20.6% of the control population without a history of warts or with short-duration wart disease. Thus, our results indicate that the epidemiology, as well as morphology, of common warts is closely linked to the virus type. PMID- 9209680 TI - Opposite regulation of tyrosinase and glutathione peroxidase by intracellular thiols in human melanoma cells. AB - Conditions of oxidative stress lead to down-regulation of glutathione (GSH) and glutathione peroxidase (GPO), which could be responsible for tyrosinase induction in pigment cells. To address this question, the effects of selective modulation of GSH metabolism on melanogenic parameters of slightly and highly melanized melanoma cells were examined. Under standard culture conditions (100 microM cystine, 100 microM tyrosine), the levels of GSH and the activities of glutathione reductase (GR) and GPO were found to be directly related to the pigmentation of melanoma cells. Exposure to 50 microM buthionine sulfoximine for 72 h decreased tyrosinase activity by 30-50% and GSH levels by more than 95%. In contrast, inhibition of GR activity with bis(chloroethyl)nitrosourea or stimulation of GPO activity with sodium selenite did not affect tyrosinase activity nor pigment formation in the melanoma cells tested. Since cysteine (CysH) is a precursor of the GSH tripeptide, the modulation of tyrosinase and GPO activity by the extracellular cystine concentration was also examined. When the cystine concentration was increased from 0 to 200 microM, a dose-dependent decrease in tyrosinase activity was associated with dose-dependent increases in GPO activity and in cell levels of CysH and GSH. The results indicate that cellular thiols coregulate the activities of tyrosinase and GPO in opposite directions. These interdependent processes could provide melanoma cells with protection against oxidative stress at low as well as at high thiol concentration. PMID- 9209681 TI - Human melanoma cells generate leukotrienes B4 and C4 from leukotriene A4. AB - We examined the synthesis of leukotrienes (LTs) in human melanoma cells in order to assess the function of LTs in human melanocytes. LTA4 hydrolase, which catalyzes the conversion of LTA4 to LTB4, was detected in the supernatant of cultured human melanoma (MeWo) cells and melanoma cells obtained from patients. Immunoblotting analysis using an antihuman LTA4 hydrolase antibody showed LTA4 hydrolase to be a 70-kDa protein in both MeWo and melanoma cells. Considerable activity of LTC4 synthase, which catalyzes the conversion of LTA4 to LTC4, was detected in the microsomal fraction of both MeWo and melanoma cells. The HPLC profile of the LTC4 synthase reaction products revealed that LTC4 was the main product. LTD4 was not detected under these conditions, indicating that the microsomal fraction of human melanoma cells lacks the membrane-bound gamma glutamyl transferase that converts LTC4 to LTD4. LTC4 synthase activity was inhibited by the addition of MK-886, and was not altered by treatment with N ethylmaleimide or 1-chloro-2,4-dinitrobenzene. These results indicate that the enzyme responsible for the conversion of LTA4 to LTC4 in human melanoma cells is LTC4 synthase rather than a nonspecific or microsomal glutathione-S-transferase. These results also suggest that human melanoma cells can generate LTB4 and LTC4 from LTA4, and that this process is catalyzed by two enzymes: LTA4 hydrolase and LTC4 synthase. PMID- 9209682 TI - Stable expression of CD1a molecule in human epithelial cell lines shows temperature-dependent expression and affects cell morphology and growth. AB - The human CD1a molecule is a transmembrane protein which shares structural similarities with HLA class I molecules. It has restricted tissue distribution in normal individuals, and is a useful diagnostic marker for certain disease states such as Langerhans cell histiocytosis. In order to investigate the function of this molecule, a cDNA fragment encoding the CD1a molecule was cloned into several EUKARYOTIC expression vectors which were then used to establish human epithelial cell lines stably expressing the membrane-bound CD1a molecule. Human keratinocytes (HaCaT) and epithelial cells (HeLa) stably expressing CD1a were established by retroviral-mediated gene transfer and DNA transfection, respectively. Expression and localization of the CD1A molecule were then confirmed by Northern blot analysis and immunofluorescence methods. CD1a expression appears to have profound effects on cellular growth and morphology. Both stably CD1a-expressing HeLa and HaCaT cells showed increased doubling times, and up to 20% of CD1a-expressing cells showed altered cell morphology. Clonogenicity experiments demonstrated a reduction in colony size and plating efficiency was augmented in CD1a-positive cells when compared with vector transfected/infected controls. Our findings suggest that CD1A expression may act as a negative growth regulator in these cells in vitro. Furthermore, lower temperatures greatly enhanced the expression of CD1a at both the protein and mRNA levels in a time-dependent fashion. Since the physiological skin temperatures lie well below the core temperature, this observation may have important implications in the study of Langerhans cells in vitro. PMID- 9209683 TI - Subcellular distribution of 220 kDa antigen in the intercellular spaces of normal human epidermis. AB - We generated a monoclonal antibody, 7C1, to an extract of pig snout skin used as antigen. The antigen for 7C1 was identified by immunoblotting as a 220 kDa epidermal protein. It was found immunologically to be distributed mainly in the intercellular spaces throughout the living cell layers of normal human epidermis, but its distribution changed in some keratinizing disorders. Study of the association of this protein with subcellular structures by immunoelectron microscopy of normal human skin revealed that it was localized in (1) lamellar granules, (2) intercellular spaces, especially in the lamellar structures derived from the lamellar granules, (3) small vesicles near the cell boundaries and (4) small vesicles in the Golgi apparatus. These results suggest that in humans the 220 kDa protein originates in the Golgi apparatus and is secreted via either the lamellar granules or the small secretory vesicles into the intercellular spaces of the epidermis, where it may be involved in the epidermal permeability barrier function and possibly in keratinizing differentiation. PMID- 9209684 TI - Upregulation of vitamin D receptor levels by 1,25(OH)2 vitamin D3 in cultured human keratinocytes. AB - The natural biologically active form of vitamin D3, 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), possesses antiproliferative, prodifferentiating and immunomodulatory properties. The actions of 1,25(OH)2D3 are mediated through the intracellular vitamin D receptor (VDR), and the level of VDR is believed to determine the cellular responsiveness to vitamin D3. In the present study we examined the effects of 1,25(OH)2D3 on the expression of VDR and its message in cultured human keratinocytes. Western analysis showed the mean VDR content to be higher in undifferentiated cultures (175 pg/microgram protein) than in differentiated cultures (90 pg/microgram protein). Incubation with 1,25(OH)2D3 induced an increase in the VDR in both undifferentiated and differentiated keratinocytes. The VDR increase was detectable after 2 h and maximal (approximately two-fold stimulation) after 8 h. The 1,25(OH)2D3-induced stimulation of VDR levels was dose dependent with a maximum at 10(-7) M. The VDR mRNA levels as determined by the ribonuclease protection assay showed a peak (50% stimulation) after approximately 2 h. Although this increase in VDR mRNA was not statistically significant, the results indicate that the ligand-induced upregulation of VDR involves increased transcription. The upregulation of VDR levels may increase the responsiveness to 1,25(OH)2D3 and may, therefore, be an important mechanism for regulating the effects of 1,25(OH)2D3 on keratinocyte proliferation and differentiation. PMID- 9209685 TI - Changes in protein and mRNA levels of growth factor/growth factor receptors in rat livers after administration of phenobarbitone or methylclofenapate. AB - The effects of phenobarbitone and methylclofenapate were studied on the expression of growth factor and growth factor receptors in livers of male Wistar rats. The major findings were: (1) a significant reduction in epidermal growth factor receptor (EGFR) protein observed with both treatments, and (2) levels of EGFR transcripts were only slightly decreased with both compounds. The reduction in the receptor level therefore does not occur via regulation of transcription. Mannose-6-phosphate receptors (M6PR, also called insulin-like growth factor II receptor) and M6PR transcripts remained unchanged in both experimental groups. Hepatocyte growth factor receptor (HGFR) transcripts were also unchanged in both experimental groups. Transcript levels of transforming growth factor-beta 1 (TGF beta 1) were lower in both treatment groups compared with the control; the reduction was significant in the methylclofenapate group. This may have relevance to the finding by others that nafenopin, another peroxisome proliferator, suppresses rat hepatocyte apoptosis. Another finding of general interest was that the three "housekeeping genes", namely albumin, actin and glyceraldehyde-3 phosphate dehydrogenase, were influenced by both treatments thus limiting their use as controls for gel loading. The adaptation of a growth regulatory mechanism via EGFR and its ligands may provide conditions such that cells with aberrant growth control have a selective growth advantage over normal cells thus promoting tumorigenesis. PMID- 9209686 TI - Congener specific effects by polychlorinated biphenyls on catecholamine content and release in chromaffin cells. AB - The effects of the non-planar polychlorinated biphenyl (PCB) congener 2,2',4,4' tetrachlorobiphenyl (2,4-TCB) and of the coplanar PCB congener 3,3',4,4' tetrachlorobiphenyl (3,4-TCB) were investigated on the catecholamine content and release from bovine adrenal chromaffin cells in culture. Each congener was tested at three concentrations (20, 50 and 100 microM) and two exposure periods (24 h and 5 days). Catecholamine release induced by K(+)-stimulation as well as catecholamine content of Triton X-100 treated cell cultures were examined using high-performance liquid chromatography (HPLC). 2,4-TCB showed dose- and time dependent effects. 2,4-TCB at 100 microM reduced the K(+)-stimulated catecholamine release after 24 h of exposure. After 5 days of exposure, 2,4 TCB at 50 and 100 microM drastically reduced the K(+)-stimulated catecholamine release. 3,4-TCB even at a concentration of 100 microM over exposure of either 24 h or 5 days had no effects on the K(+)-stimulated secretion. When chromaffin cells, exposed to 2,4-TCB, were lysed with 0.5% Triton X-100, a dose- and time dependent reduction of the catecholamine content appeared. The 3,4-TCB did not reduce the catecholamine content. Conversely there seemed to be a trend towards an increase in catecholamine content. Spontaneous release of catecholamines was strongly increased by the non-planar 2,4 TCB, while the coplanar 3,4 TCB showed no effects on this parameter. Furthermore, the effects of 2,4 TCB appeared to be reversible after replacing the highest concentration (100 microM) of the TCB solution with culture-medium at the end of the 24-h exposure. Thus, K(+) stimulated catecholamine release and the catecholamine content of bovine adrenal chromaffin cells was effectively reduced by the non-planar PCB congener whereas spontaneous catecholamine release was strongly increased. The coplanar PCB congener was ineffective at the same conditions. PMID- 9209687 TI - The response of hepatocytes isolated from phenobarbitone treated mice to mitogenic growth factors. AB - The ability was investigated of epidermal growth factor (EGF) and hepatocyte growth factor (HGF) to stimulate DNA synthesis in hepatocytes isolated from C57B1/6J mice following 1, 3, 7, 30 and 90 days pre-treatment with the hepatomegalic drug, phenobarbitone (PB). A 3-fold increase in S-phase labelled hepatocytes was observed in the absence of growth factors after 3 days treatment with PB, which was not seen at other investigated time points. This suggests that the proliferative influence present in vivo at this time interval is maintained in the ex vivo model. Maximum labelling indices of > 5-fold the unstimulated control value were observed in hepatocytes isolated from control and 1 day PB pre treated mice when cultured in the presence of 5 or 10 ng/ml EGF or HGF. Hepatocytes isolated from 3, 7, 30 or 90 day treated mice showed a considerably reduced responsiveness to growth factors; maximum labelling indices did not exceed by a factor of 2 the value obtained in the absence of growth factors. However, the apparent decrease in responsiveness to growth factors in hepatocytes isolated from 3 day pre-treated mice was due to an increased background level of proliferation and the attainment of a 'ceiling level' of DNA synthesis at approx. 35%. DNA synthesis was not further enhanced by addition of both EGF and HGF. This maximal level of stimulation may indicate that only a specific hepatocyte sub population is capable of responding to growth factors under the conditions employed. The loss in sensitivity to mitogenic stimuli after 7 days PB pre treatment correlates with a reported decrease in receptor protein and mRNA levels in rats and coincides with the in vivo shift from hyperplasia to hypertrophy. PMID- 9209688 TI - d-Amphetamine-induced hepatotoxicity: possible contribution of catecholamines and hyperthermia to the effect studied in isolated rat hepatocytes. AB - Amphetamines are indirect-acting sympathomimetic drugs widely abused due to their physical and psychostimulating effects. However, the use of these drugs has been associated with numerous reports of hepatotoxicity. While glutathione depletion induced by amphetamines contributes to the exposure of hepatocytes to oxidative damage, other indirect effects attributed to amphetamines may have a role in cell injury. To examine this possibility, Wistar rats were used for plasma measurements of d-amphetamine and catecholamines (noradrenaline, adrenaline and dopamine) (15 min) after i.p. injection of d-amphetamine (5, 20 and 80 mg/kg). Freshly isolated rat hepatocytes were put into contact for 2 h with concentrations of d-amphetamine and catecholamines similar to those found in vivo. Since hyperthermia is a common consequence of acute amphetamine intake, the study using isolated hepatocytes was conducted at 37 degrees C and also at 41 degrees C in order to simulate high temperature levels. We found that hyperthermia was an important cause of cell toxicity: in vitro, a rise in incubation temperature from 37 to 41 degrees C causes oxidative stress in freshly isolated rat hepatocytes, as shown by a depletion of reduced glutathione (GSH; 23%), an increase of oxidized glutathione (GSSG; 157%), the induction of lipid peroxidation with 77% increase of thiobarbituric acid substances TBARS) and the consequent loss of cell viability (< or = 44%). Single treatment of isolated hepatocytes with catecholamines at 37 degrees C induced lipid peroxidation (29% increase of TBARS) but had no effect on glutathione or cell viability. Conversely, a single treatment with d-amphetamine induced glutathione depletion (< or = 24% depletion of GSH) with no effect on lipid peroxidation or cell viability. Also, d-amphetamine potentiated the induction by catecholamines of lipid peroxidation at 37 degrees C (< or = 48% increase of TBARS), while concomitant treatment of d-amphetamine and catecholamines potentiated cell death at 41 degrees C (< or = 56% of cell death) although no effect on viability was seen at 37 degrees C. It is concluded that the aforementioned modifications induced by d-amphetamine in vivo are cytotoxic to freshly isolated rat hepatocytes. PMID- 9209689 TI - Implication of CYP 3A in the toxicity of cyclosporin G (CsG), cyclosporin A (CsA) and FK506 on rat hepatocytes in primary culture. AB - FK506, cyclosporin A (CsA), and its structural analog cyclosporin G (CsG) are immunosuppressant drugs mainly metabolized by hepatic cytochrome P-450 3A (CYP 3A) oxygenase. FK506 metabolites exhibit greater toxicity than the parent drug, while CsA metabolites are far less toxic than CsA itself. The aim of our study was to compare the toxicity of CsG with CsA and FK506 as a function of CYP 3A induction. Hepatocytes from Wistar rats with or without dexamethasone (DEX) induction (200 mg/kg per day, p.o for 4 days) were used in primary culture. The DEX-inductive effect on CYP 3A was assessed by SDS-PAGE. After 6 h incubation with CsG, CsA or FK506 (5 to 200 microM), cell viability (expressed as IC50), intracellular calcium content and apoptosis were evaluated. Concerning cytotoxicity, IC50 values for CsG, CsA and FK506 were 75, 50 and 180 microM respectively in non-induced cultures, and 150, 120 and 25 microM in induced cultures. For intracellular calcium content, a dose-dependent increase was observed in all cultures. However this increase is more important for CsG and CsA in non-induced cultures (150%) compared to induced cultures (110%) at 150 microM. Conversely for FK506, this increase is greater in induced cultures (150%) than in non-induced cultures (127%). Estimation of the percentage of apoptotic cells shows similar variations. Our results show that the toxicity of the three drugs in rat hepatocytes is dependent on CYP 3A induction: increased for FK506, decreased for CsA and CsG. Moreover, with regard to the three tests used, the toxic effects of CsG are close to those of CsA, indicating that CsG metabolites are also less toxic than the parent drug. PMID- 9209690 TI - Influence of subchronic administration of oestradiol, ethinyloestradiol and oestradiol sulphamate on bile flow, bile acid excretion, and liver and biliary glutathione status in rats. AB - The cholestatic effect of the newly developed oestradiol sulphamate (J995) was compared with that of the natural oestrogen oestradiol (E) and of the cholestatic standard oestrogen ethinyloestradiol (EE). Female ovariectomized rats were orally treated for 7 days with oestrogen doses molar equivalent to 0.01, 0.1, 1, and 10 mg E/kg body wt. Bile flow, biliary bile acid and glutathione excretion as well as liver glutathione status were determined after bile duct cannulation under the influence of ketamine anaesthesia. For systemic oestrogen activity, the increase of uterine weight was measured. J995 showed the highest oestrogenic activity followed by EE and E. Impairment of bile flow and biliary glutathione excretion (GSH, GSSG) were found to be in the order E < J995 < EE. As all three oestrogens did not influence bile acid excretion, we postulate that mainly the bile acid independent fraction of bile flow was inhibited, caused at least partly by impairment of canalicular glutathione transport. Based on the results of these studies, we conclude that a tenfold higher dose of J995 exhibited the same cholestatic effect as EE. Together with higher systemic oestrogenic activity, J995 may be used as a prodrug with reduced hepatic side-effects to replace EE in contraception strategies. PMID- 9209691 TI - DNA binding activity of the mammalian translation elongation complex: recognition of chromium- and transplatin-damaged DNA. AB - The elongation factor complex, EF-1H, serves an essential function in protein biosynthesis in eukaryotic cells, although the role of EF-1H in other physiological processes is unknown. In this report, we demonstrate that three components of EF-1H (EF-1 beta, EF-1 delta, EF-1 gamma) bind to DNA modified with chromium (Cr), a potent DNA-damaging agent and an established human carcinogen. The EF-1H complex also binds to transplatin modified DNA but not to cisplatin modified DNA. These results demonstrate that the EF-1H complex has functional DNA binding activity and is capable of recognizing the distortions in DNA structure resulting from the covalent binding of Cr and transplatin to DNA. PMID- 9209692 TI - Low levels of cadmium chloride damage the corneal endothelium. AB - The effect of cadmium chloride on the integrity of the endothelium of isolated bullfrog (Rana catesbeiana) corneas was examined by spectrophotometric analysis of corneal uptake of the vital stain Janus green and by scanning electron microscopy (SEM). The uptake of Janus green by the endothelium was dose related between 1.0 and 100.0 microM CdCl2. The effect of cadmium was significantly attenuated by the calcium channel blocker SKF 96365 and was augmented by the calcium ionophore A23187, indicating that cadmium influx through calcium channels is an important determinant of its cellular effect. The effect of cadmium was not altered by changes in the external calcium concentration, indicating that the mechanism does not involve competitive inhibition by calcium. SEM demonstrated significant structural damage to the corneal endothelium exposed to cadmium chloride, including focal disruption and denuding of the apical endothelial membrane. PMID- 9209693 TI - Effects of dose on the methylation of selenium to monomethylselenol and trimethylselenonium ion in rats. AB - Mechanisms and metabolic significance in rats of methylation to the reduced form of selenium (Se), i.e., selenide (Se2-), were studied by dose- and time-related experiments with injection of selenite. Urinary Se-metabolites were determined by HPLC using an inductively coupled argon plasma-mass spectrometer as an in-line detector (HPLC/ICP-MS method). Although only monomethylselenon (MMSe) has been detected in urine of normal rats even in those fed a Se-excess diet, the three types of Se-metabolites - MMSe, trimethylselenonium ion (TMSe), and inorganic Se, were detected in urine of Wistar rats injected with selenite (0, 0.1, 0.3, 0.5 and 1.0 mg Se/kg body weight) into the tail vein. The amount of the three Se metabolites was plotted against the total urinary Se concentration and shown to change dose- and time-dependently. The monomethylated metabolite, i.e., MMSe, increased in urine rapidly at first and was slowly followed by linear dose dependent excretion of the trimethylated metabolite, TMSe. The new methylation pathway of MMSe leading to TMSe was assumed to be induced or activated when the dose of Se exceeds the limit of the normal capacity for monomethylation. Progressive methylation reactions were suggested to be regulated enzymatically. PMID- 9209694 TI - Antidotal efficacy of quinuclidinium oximes against soman poisoning. AB - The efficiency of newly synthesized oxime derivatives of quinuclidinium were tested in vitro on soman inhibited acetylcholinesterase (AChE) of human erythrocytes and in vivo using soman poisoned mice. For this purpose, the inhibitory power of oximes (IC50), acute toxicity (LD50) as well as reactivating and protective capacities with respect to soman-inhibited AChE were determined for each of the oximes. All oximes tested were ineffective in vitro but protected mice very efficiently (BM-1 protects against 4LD50 of soman). The results indicate that the in vivo effectiveness of quinuclidinium oximes against soman poisoning may not be related to reactivation or protection of AChE but rather to some other mechanism of the cholinergic system. PMID- 9209695 TI - In vitro evidence for a Donnan distribution of Mg2+ and Ca2+ by chondroitin sulphate in cartilage. AB - Fluoroquinolones are known for their ability to form chelate complexes with magnesium. Cartilage lesions observed in juvenile animals after quinolone treatment very probably are a consequence of the lack of functionally available magnesium. In cartilage, which contains high amounts of negatively charged proteoglycans, a Donnan distribution can be expected leading to an inhomogeneous distribution of ions (such as magnesium), which may support the toxic effects of magnesium deficiency or quinolone treatment of cartilage. We performed in vitro experiments using dialysis tubes to simulate the unequal distribution of proteoglycans in cartilage and measured the distribution of magnesium, calcium and ofloxacin. We found that the concentration of free magnesium is significantly reduced with the chondroitin sulphate-free solution due to a Donnan effect. For example, using a 3% chondroitin sulphate solution (outside the tubing) dialysed against a chondroitin sulphate-free solution (inside the tubing) the magnesium concentration decreased by 24% from 0.55 +/- 0.02 to 0.42 +/- 0.04 mmol/l inside the tubing during 48 h observation (P < 0.01). Under physiological conditions this unequal distribution of magnesium probably will be much more pronounced because chondroitin sulphate concentrations in cartilage are higher; nevertheless, magnesium concentration is sufficient for regular function of the tissue. During the sensitive phase of quinolone toxicity, magnesium in juvenile cartilage is lower than at other time points during postnatal development. Moreover, additional complexation by quinolones may further reduce the concentration of functionally available magnesium below the critical level. PMID- 9209696 TI - Hypolipidemic effect of Curcuma comosa in mice. AB - The hypolipidemic effect of an ethyl acetate extract of the rhizome of Curcuma comosa Roxb was investigated in mice. Intragastric administration of the extract significantly decreased plasma lipid levels of both triglyceride and cholesterol but increased liver triglyceride content. Liver weight and plasma activities of glutamic oxaloacetic transaminase and glutamic pyruvic transaminase were not affected by a single administration. Prolonged treatment did not further decrease plasma lipid level but caused further increases in liver triglyceride content and weight. The lower plasma cholesterol activity of C. comose extract was found to be essentially associated with elevation of plasma HDL cholesterol level, increased excretion of fecal cholesterol and bile salt. The increase persisted throughout the period of treatment. These results suggest that C. comosa has hypolipidemic action. It exerts hypocholesterolemic activity by accelerating mobilization of cholesterol from peripheral tissues into liver and enhancing excretion of cholesterol and bile salt into feces. PMID- 9209697 TI - Effects of highly purified eicosapentaenoic acid on vascular reactivity to angiotensin II and norepinephrine in pregnant rabbits. AB - OBJECTIVE: We sought to determine whether the pressor response to an infusion of angiotensin II during pregnancy would be reduced by the oral administration of highly purified eicosapentaenoic acid ethyl ester (EPA-E). METHODS: We administered EPA-E orally to nine pregnant rabbits (200 mg/kg/day) from day 5 of gestation to day 5 postpartum. Pressor responses to graded doses of intravenously infused angiotensin II and norepinephrine were examined serially during pregnancy and the postpartum period. RESULTS: The EPA-E-treated, as well as the control pregnant rabbits (n = 6), were significantly less responsive to the vasoconstrictor effects of angiotensin II throughout pregnancy than the nonpregnant rabbits (n = 8). The vascular reactivity to infused angiotensin II in EPA-E-treated pregnant rabbits, as compared with that of control pregnant rabbits was unchanged, but reactivity to angiotensin II was lower only on the fifth postpartum day. The pressor responses to infused norepinephrine were unchanged during pregnancy and in the postpartum period, and EPA-E did not alter the response. The litters of four of the nine rabbits were partially or completely macerated. CONCLUSION: EPA-E did not reduce the already blunted pressor responsiveness to angiotensin II during pregnancy in rabbits. The effect of EPA-E in a state of increased pressor responsiveness during pregnancy due to a deficiency of vasodilatory prostaglandins needs to be determined. PMID- 9209698 TI - The distribution of centrosomes in endothelial cells of the rat aorta and inferior vena cava. AB - Centrosomes are preferentially oriented toward the heart in endothelial cells (ECs) of the pig aorta and pig and rabbit inferior vena cava (IVC). In the rabbit aorta this preferential orientation of the centrosome toward the heart decreases with age. To determine if this is also true in the rat, a species which is more amenable to experimental manipulation than the pig or the rabbit, we determined the position of centrosomes relative to the nucleus in ECs lining the aorta and IVC using whole mounts of vessels that were immunofluorescently stained with sera specific for centrosomes. In both the thoracic and abdominal aorta of the rat the majority of the ECs (60%) had centrosomes on the heart side of the nucleus, 25% had centrosomes on the side of the nucleus away from the heart and 15% had centrosomes in a central position in the cell. Similar results were obtained in the IVC of the rat where these values were, 58%, 31% and 11% respectively. A comparable preferential orientation of centrosomes toward the heart was also seen in the ECs of thoracic and abdominal aortas and IVCs of weanling and young adult rats and this did not decrease with age as it does in the rabbit aorta. When segments of the rat aorta were placed in organ culture, the percentage of ECs with preferentially oriented centrosomes decreased by 48 hrs, even though the cells remained elongated in shape. We have recently demonstrated that ECs in the rat aorta are normally migrating in the direction of the heart and thus in the direction in which the centrosomes in rat aortic ECs are preferentially oriented. This correlation is consistent with the general hypothesis that the centrosome position defines the direction of migration in monolayers of cells. PMID- 9209699 TI - Dietary conjugated linoleic acid reduces plasma lipoproteins and early aortic atherosclerosis in hypercholesterolemic hamsters. AB - Conjugated linoleic acid is a collective term used to designate a mixture of positional and geometric isomers of linoleic acid in which the double bonds are conjugated. Unlike linoleic acid, there is a paucity of information regarding the effect of dietary conjugated linoleic acid on plasma lipoproteins and aortic atherosclerosis. Therefore, fifty hamsters were divided into five groups of ten and fed 0 (Control), 0.06 (LOW), 0.11 (MEDIUM), and 1.1 (HIGH) en% conjugated linoleic acid or 1.1 en% linoleic acid. Blood samples were taken at 4, 8 and 11 weeks for plasma lipid analyses and for plasma tocopherol assay at sacrifice. Animals fed the conjugated linoleic acid-containing diets collectively had significantly reduced levels of plasma total cholesterol, non-high density lipoprotein cholesterol, (combined very low and low density lipoprotein) and triglycerides with no effect on high density lipoprotein cholesterol, as compared to CONTROLs. Linoleic acid-fed animals relative to CONTROLs also had reduced plasma total cholesterol, non-high density lipoprotein cholesterol and triglycerides, but only the latter was statistically significant. Compared to the CONTROL group, plasma tocopherol/total cholesterol ratios determined from plasma pools for the LOW, MEDIUM and HIGH conjugated linoleic acid and linoleic acid groups were increased by 48%, 48%, 86% and 29%, respectively, suggesting a tocopherol-sparing effect, at least for the conjugated linoleic acid treatment. Morphometric analysis of aortas revealed less early atherosclerosis in the conjugated linoleic acid and linoleic acid-fed hamsters compared to the CONTROL group. PMID- 9209700 TI - Potential advantages of treatment of transplanted saphenous vein aorto-coronary artery bypass grafts with beta irradiation to prevent graft occlusion. AB - Intimal proliferation or Neointimal hyperplasia (NIH) is a vascular lesion that often arises in arteries after balloon angioplasty or other vessel wall injuries. FIH is a vascular lesion that develops in autologous saphenous vein grafts (SVG) after transplantation into the aorto-coronary circulation or the peripheral vascular circulation. FIH shares elements of smooth muscle migration, proliferation and fibrous tissue deposition in common with nibrointimal proliferation (NIH). Either NIH of a coronary artery or FIH of a SVG obstruct the vascular lumen and result in myocardial dysfunction. Local radiotherapy has been used for several decades to reduce the post-operative recurrence of the fibrovascular proliferations of pterygia and keloids. Similarly, in animal and human experiments, endovascular radiotherapy has been shown to reduce arterial smooth muscle proliferation. Consideration of the similarities of vascular smooth muscle cell proliferation in NIH and FIH leads one to suggest that endovascular beta irradiation can reduce FIH as well as it reduces NIH. The goal of such treatment is to achieve a clinically significant decrease in the morbidity and mortality resulting from SVG occlusions. The potential for large reduction of the consequences of SVG occlusion, the very large number of patients at risk, and the simplicity of the proposed intervention encourages prompt scientific evaluation of this technique. PMID- 9209701 TI - Peroxisome proliferators and peroxisome proliferator activated receptors (PPARs) as regulators of lipid metabolism. AB - Peroxisome proliferation (PP) in mammalian cells, first described 30 years ago, represents a fascinating field of modern research. Major improvements made in its understanding were obtained through basic advances that have opened up new areas in cell biology, biochemistry and genetics. A decade after the first report on PP, a new metabolic pathway (peroxisomal beta-oxidation) and its inducibility by peroxisome proliferators were discovered. More recently, a new type of nuclear receptor, the peroxisome proliferator-activated receptor (PPAR), has been described. The first PPAR was discovered in 1990. Since then, many other PPARs have been characterized. This original class of nuclear receptors belongs to the superfamily of steroid receptors. With activation of cell signal transduction pathways, the occurrence of PPARs provides, for the first time, a coherent explanation of mechanisms by which PP is triggered. Nevertheless, although many compounds or metabolites are capable of activating PPARs, the natural direct ligands of these receptors have not been, up to now, clearly identified, with, however, the exception of 15-deoxy-12,14-prostaglandin J2 which is the ligand of PPAR gamma 2 while leukotrien LTB4 binds PPAR alpha. At this stage, the hypothesis of some orphan PPARs (ie receptors without known ligand) can not be ruled out. Despite these relatively restrictive aspects, the mechanisms by which activation of PPARs leads to PP become clear; also, coherent hypotheses among which a scenario involving receptor phosphorylation or a heat shock protein (ie HSP 72) can be proposed to explain how PPARs would be activated. The aim of this note is to review recent developments on PPARs, to present members up to now recognized to belong to the PPAR family, their characterization, functions, regulation and mechanisms of activation as well as their involvement in lipid metabolism regulation such as control of beta-oxidation, ketogenesis, fatty acid synthesis and lipoprotein metabolism. As an introducing section, a brief review of the major events between the first report of PP in mammals and the discovery of the first PPAR is given. Another section is devoted to current hypotheses on mechanisms responsible for PPAR activation and PP induction. Rather than an exhaustive presentation of cellular alterations accompanying PP induction, a dynamic overview of the lipid metabolism is provided. By assessing the biological significance of this organellar proliferative process, the reader will be led to conclude that the discovery of PPARs and related gene activation through peroxisome proliferator responsive element (PPRE) makes PP induction one of the most illustrative examples of control that occurs in lipid metabolism. PMID- 9209703 TI - Peroxisome proliferator-activated receptor (PPAR)-dependent and -independent transcriptional modulation of several non-peroxisomal genes by peroxisome proliferators. AB - To better characterize peroxisome proliferator-induced non-peroxisomal responses, we searched the mRNAs of which the levels were modulated by proliferators. We used the PCR-based methods including differential display. The mRNAs were divided into at least four groups by their time-courses of induction and repression: group 1 very rapidly increased then decreased; group 2 increased after a time lag (well-characterized peroxisomal mRNAs belonged to this group); group 3 decreased reciprocally compared with group 2 mRNAs; group 4 increased after group 2 mRNAs, with a much longer lag period. All of these modulations cannot be explained by peroxisome proliferator action through PPAR and RXR dimerization on the target genes to activate transcription. Another unidentified transcription factor may be involved in some of these modulations. It will also be important to consider PPAR independent pathways when studying the diverse effects of peroxisome proliferators. PMID- 9209702 TI - The effects of fibrates and thiazolidinediones on plasma triglyceride metabolism are mediated by distinct peroxisome proliferator activated receptors (PPARs). AB - The hypolipidemic fibrates and antidiabetic thiazolidinediones display potent triglyceride-lowering activities. Studies on the molecular action mechanisms of these compounds indicate that thiazolidinediones and fibrates exert their action by activating distinct transcription factors of the peroxisome proliferator activated receptor (PPAR) family, resulting in increased expression of lipoprotein lipase (LPL) and decreased expression of apolipoprotein (apo) C-III, both key-players in plasma triglyceride metabolism. Fibrates, on the one hand, are PPAR alpha activators, which selectively induce LPL mRNA levels and activity in the liver. Furthermore, hepatic apo C-III mRNA levels and protein production strongly decrease after fibrate treatment. On the other hand, thiazolidinediones, which are high affinity ligands for PPAR gamma, have no effect in the liver, but act primarily on adipose tissue, where they induce LPL mRNA levels and activity. The modulation of the expression of the LPL and apo C-III genes in liver and adipose tissue is correlated with the tissue-specific distribution of the respective PPARs (PPAR gamma expression being restricted to adipose tissue, whereas PPAR alpha is expressed predominantly in liver) confirming that fibrates and thiazolidinediones exert their effects primarily through PPAR alpha and PPAR gamma respectively. This distinct tissue-specific transcriptional regulation of genes involved in lipid metabolism by fibrates and thiazolidinediones indicates that research of compounds displaying combined PPAR alpha and PPAR gamma activation potential should lead to the discovery of more potent triglyceride lowering drugs, which may be of use in the treatment of hypertriglyceridemia. PMID- 9209705 TI - PPAR gamma and the control of adipogenesis. AB - We recently cloned PPAR gamma as a factor that binds to an enhancer which has specificity for adipose cells. When expressed ectopically, PPAR gamma converts fibroblasts into bona fide preadipose cells. Upon application of activators or PPAR gamma ligands, these cells differentiate into fat cells. Most recently, we have been trying to understand the nature of natural ligands that activate PPAR gamma and the protein domains that control adipogenesis. With regards to ligands, we have shown that an unusual prostanoid, 15-deoxy delta 12,14PG J2, can bind to PPAR gamma and activate it. A second transcription factor that is induced early in differentiation, ADD1/SREBP1, appears to promote the formation of PPAR gamma ligands. Transfection of this molecule, a member of the bHLH family, causes the secretion of molecules that can serve as ligands for PPAR gamma. This ligand-like activity is specific for the gamma isoform of PPAR. Current studies are attempting to identify these potentially novel ligands. With regard to structure function of PPAR gamma, we first analyzed the adipogenic activity of the three isoforms of PPAR: alpha, gamma and delta. Using appropriate activators of each it is clear that PPAR gamma has the most adipogenic action. PPAR alpha can be adipogenic with high levels of the strongest activators and PPAR delta does not stimulate fat cell differentiation. To identify the domain(s) of PPAR gamma responsible for differentiation, chimeras between PPAR gamma and PPAR delta were created and transfected into fibroblasts. This has allowed the isolation of relatively small regions of this molecule that are responsible for differentiation. PMID- 9209706 TI - Transcriptional regulation by glucose in the liver. AB - Numerous hepatic and adipocytic genes are transcriptionally controlled by glucose and insulin. It is the case, for example, of the pyruvate kinase L (L-PK) gene in the liver and of the spot 14 gene in adipocytes, coding for proteic factors of glycolysis and lipogenesis, respectively. At the hepatic level, the role of insulin is mainly to stimulate the synthesis of glucokinase, needed for phosphorylation of glucose to glucose 6-phosphate. An efficient regulation of the L-PK gene by glucose also needs the synthesis of the glucose transporter (Glut2): in its absence, transcription of the gene is independent of the presence of glucose in the medium. The role of Glut2 can be to enhance the depletion of gluconeogenic cells into glucose-6-phosphate (G6-P) when cultivated without glucose. G6-P seems to act by one of its metabolites in the pentose phosphate pathway, probably a pentose phosphate, maybe xylulose 5-phosphate. The active metabolites of this pathway could control the activity of protein kinase and protein phosphatase cascades, leading to a modification of the phosphorylation state of the glucose response complex. This complex is assembled by so-called glucose/carbohydrate response elements (GIRE, ChoRE) that are composed of E boxes of the CACGTG type, more or less modified, forming a palindrome whose both parts are separated by five bases. These sequences are able to bind USF1 and USF2 proteins, which seem to be necessary to the glucose response. However, the binding of USF proteins to the GIRE of the L-PK gene, appreciated by in vivo footprints, is not modulated by nutritional conditions. Therefore, these USF proteins could interact with different partners which are targets of regulating cues: transcription factors bound in the immediate vicinity of the glucose response complex, notably the HNF4 factor, and, maybe, other proteins interacting with the USF factors assembled to the GIRE. The actually ongoing experiments try to appreciate the nature and the role of these partners, and to evaluate the metabolic response of mice whose USF genes were disabled by homologous recombination. PMID- 9209704 TI - Retinoid X receptor (RXR alpha) gene expression is regulated by fatty acids and dexamethasone in hepatic cells. AB - This work describes the molecular mechanism of fatty acid and hormonal modulation of retinoid X receptor (RXR alpha) in rat liver. We examined the effects of different fatty acids (myristic-, stearic-, linolenic-, oleic-, arachidonic- and tetradecylthioacetic acid (TTA)) and the synthetic glucocorticoid dexamethasone on RXR alpha mRNA and protein steady-state levels in hepatoma cells and cultured hepatocytes. Fatty acids induced the RXR alpha gene expression where TTA showed the most inductive effect (three-fold induction). Dexamethasone alone resulted in a stronger induction (up to seven-fold in hepatocytes), and in combination with fatty acids, an additive or synergistic effect was observed. The RXR alpha protein level in cultured hepatocytes showed a similar pattern of regulation, with a slight inductive effect of fatty acids and an additive or synergistic effect was observed in combination with dexamethasone. Our results indicate that the RXR alpha gene expression is under distinct regulation by fatty acids and dexamethasone acid which strongly suggests a coupling with the lipid metabolizing system and the retinoid signaling pathway. PMID- 9209707 TI - Glucose-dependent transcriptional regulation of the human sucrase-isomaltase (SI) gene. AB - We have previously shown that the transcription of the human sucrase-isomaltase (SI) gene was negatively regulated by glucose. Using two clonal metabolic variants of the human colon adenocarcinoma cell line Caco-2 we demonstrate here that: 1) although similar growth-related variations of phosphoenolpyruvate carboxykinase (PEPCK), frutose 1,6-diphosphatase (F1, 6-dPase), pyruvate kinase (PK) and SI mRNA levels are observed, only F1,6-dPase, PK and SI mRNA levels vary in the same way in response to modifications of glucose utilization; and 2) regulatory elements responsible for the glucose-dependent transcription of the SI gene are located within the -370/+30 region of the promoter. PMID- 9209708 TI - Glutamine and regulation of gene expression in mammalian cells. Special reference to phosphoenolpyruvate carboxykinase (PEPCK). AB - The repertoire of the actions of specific amino acids on gene expression is relatively limited in mammalian cells. Glutamine constitutes the most studied amino acid and recent works intended to demonstrate its mechanism of action on two genes: the beta-actin and the phosphoenolpyruvate carboxykinase genes. From these studies, it appears that glutamine may regulate gene expression by, at least, two different mechanisms: one through the glutamine-induced cell swelling, and another through its intracellular metabolism. The involvement of phosphatidylinositol 3-kinase in the signaling pathway triggered by cell swelling is discussed. PMID- 9209709 TI - Regulation of gene expression by fatty acids: special reference to fatty acid binding protein (FABP). AB - During the last years, the direct involvement of lipidic nutrients in the regulation of genes has been established. Fatty acids may induce or repress the transcription rate of several genes involved in both lipid and carbohydrate metabolisms. Gene up-regulation has been found in various tissues including liver, adipose tissue and small intestine. It is only triggered by saturated and unsaturated long-chain fatty acids or their CoA-derivatives. In contrast, gene down-regulation appears to be restricted to the liver. This negative effect is exerted only by polyunsaturated fatty acids. Long-chain fatty acids are able to regulate the expression of two different genes oppositely in the same cell type. The molecular mechanism of these fatty acid-mediated effects remains unclear. The involvement of members of the peroxisome proliferator-activated receptor is discussed. PMID- 9209710 TI - Fatty acid metabolism, pharmacological nutrients and hypertension. AB - The purpose of the present study was to investigate the effect of a concentrated preparation (EPA 30) containing eicosapentaenoic acid (EPA, 20:5 n-3) and docosahexaenoic acid (DHA, 22:6 n-3) on the limiting desaturation steps of the polyunsaturated fatty acid biosynthesis in spontaneously hypertensive rats (SHR). Adult SHR were divided into two groups: one group received a standard diet, and the experimental group the standard diet including 0.8% of EPA30 for 9 weeks. Blood pressure was measured at the end of the diets. The desaturase activities and fatty acid composition were determined in isolated hepatocytes. The blood pressure did not decrease in the experimental group. The desaturated products of the n-6 family (gamma-linolenic acid, 18:3 n-6 and arachidonic acid, 20:4 n-6) were lowered in the EPA30 group, when their respective substrates (18:2 n-6 and 20:3 n-6) were increased. EPA and DHA were higher in the experimental group delta 6 n-3, delta 6 n-6 and delta 5 n-6 desaturase activities were depressed approximately 20% in the EPA30 group. EPA30 being an active nutrient on the EFAs cascade, increasing the level of PG3 precursors and decreasing the level of PG2 precursors, favourable conditions have been established to reduce hypertension. The underlying mechanism related to the regulation of desaturase activities by these fatty nutrients remains to be elucidated. PMID- 9209711 TI - Recent update on the PPAR alpha-null mouse. AB - Short-term treatment of rats and mice with peroxisome proliferators (PP) results in an increase in liver peroxisome number, marked hepatomegaly and induction of several genes encoding peroxisomal and other microsomal and mitochondrial enzymes involved in fatty acid metabolism. Chronic treatment of rodents with PP results in hepatocellular carcinoma. Species differences in PP responses have been found. For example, PP such as clofibrate and gemfibrozil, are highly effective lipid and cholesterol lowering drugs in humans but do not cause peroxisome proliferation and there is no evidence for increased liver cancers in patients receiving these drugs. A receptor, designated PP-activated receptor alpha (PPAR alpha) is capable of trans-activating reporter genes containing a PP response (PPRE), but requires the presence of both PP, 9-cis retinoic acid and another receptor called RXR alpha. However, PP may not directly bind to PPAR alpha but probably indirectly disturb cellular metabolism to liberate an endogenous ligand. Subsequent to the first identification of a PPAR alpha, other members of this receptor family were found and designated PPAR alpha, PPAR beta (also called NUC1 and PPAR delta) and PPAR gamma. The alpha form is most abundant in liver and kidney, sites of peroxisome proliferation while the other two receptors are not significantly expressed in these tissues. On the basis of tissue-specific localization and spectrum of target gene activation, the physiological function of PPAR alpha and PPAR gamma appear to be related to fatty acid metabolism and regulation of adipogenesis, respectively. To gain insight into the function of PPAR alpha and its role in the peroxisome proliferator response and hepatocellular carcinogenesis, gene targeting was used to develop a PPAR alpha deficient mouse. These animals are resistant to the pleiotropic effects of PP and no induction of any known target gene has been found. Recent studies on the phenotypes of these mice have led to an understanding of the mechanism of action of PP. They have also provided a useful model to establish the physiological role of PPAR alpha in fatty acid homeostasis and inflammation. PMID- 9209712 TI - Preparation of a dansylated fibrate, a new fluorescent tool to study peroxisome proliferation. Effect on hepatic-derived cell lines. AB - The synthesis of a dansylated fibrate (DNS-X) has been performed in order to identify the cellular affinity sites of peroxisome proliferators and to establish the subcellular localization of such molecules. DNS-X has been obtained by coupling the dansy1 chloride with the amine resulting from the bezafibrate alkaline hydrolysis. The purified DNS-X has been further characterized by spectrum analysis (UV-Vis, fluorescence, [1H]/[13C]-NMR and mass). At 250 microM and incubated for 48 h with the rat hepatic derived cells (Fao cells), DNS-X stimulates 12-fold the palmitoyl-CoA oxidase, a peroxisome proliferation marker enzyme. This increase is comparable to the one obtained with well known peroxisome proliferators such as bezafibrate or ciprofibrate. The stimulation by DNS-X is specific for the overall molecule since neither the dansyl chloride, the amine, nor the precursors of DNS-X are active. The increase of palmitoyl-CoA oxidase activity is correlated with the increase of the enzyme amount as shown by immunoblotting. In agreement with the species-specificity of the fibrate neither DNS-X, bezafibrate nor ciprofibrate significantly increase palmitoyl-CoA oxidase activity and the enzyme amount in human hepatic-derived cells, HepG2. This work shows that the dansylated fibrate is a new fluorescent tool to study the subcellular localization and identification of high affinity binding sites, then further on, to elucidate the peroxisome proliferation mechanism and the action of hypolipidaemic agents of the fibrate family. PMID- 9209714 TI - Muscle-specific overexpression of human lipoprotein lipase in mice causes increased intracellular free fatty acids and induction of peroxisomal enzymes. AB - A transgenic mouse model for peroxisomal and mitochondrial induction caused by increased uptake of fatty acids in muscle was established. Transgenic mouse lines were generated using a human lipoprotein lipase (LPL) mini gene (3-20 copies) driven by the promoter of the muscle creatine kinase gene. Expression of human LPL was only observed in skeletal and cardiac muscle. In proportion to the level of LPL overexpression increased LPL activity in skeletal (up to 24-fold) and cardiac (up to three-fold) muscle, decreased plasma triglyceride levels, elevated free fatty acid (FFA) uptake by muscle tissue, weight loss (due to a reduction in muscle mass as well as adipose tissue mass) and premature death were observed. A remarkable increase in the number of mitochondria and peroxisomes was detected using oxide-electron microscopy. Proliferation of mitochondria and peroxisomes was confirmed by a dose-dependent increase of marker enzyme activity (succinate dehydrogenase and catalase). In addition, peroxisomal acyl-CoAse enzyme protein was markedly elevated whereas mRNA was increased only up to two-fold. No changes in peroxisomal proliferator activated receptor alpha mRNA was found. This degree of proliferation and enzyme activity of mitochondria and peroxisomes suggests that FFA play an important role in the induction of these organelles. In addition, myopathy characterized by excessive glycogen storage, muscle fiber degeneration, and fiber atrophy with centralization of nuclei, mimicking several forms of human myopathies was noted. Our results imply that improper regulation of muscle LPL leading to increased fatty acid levels in muscle can cause severe pathological changes. This effect may be important in the pathogenesis of human myopathies. We conclude that these transgenic mouse lines could serve as a useful animal model for the investigation of myopathies and the effects of fatty acids on the induction of mitochondria and peroxisomes. PMID- 9209713 TI - Peroxisome proliferators alter the expression of estrogen-metabolizing enzymes. AB - Exposure to some peroxisome proliferator chemicals (PPC) leads to toxic effects on sex organ function possibly by alterations of steroid hormone metabolism. A systematic search for genes whose mRNA levels are modulated by the PPC WY-14643 (WY) was carried out in rat liver, a site of steroid hormone metabolism. The sequence of one up-regulated cDNA (2480 bp) was predicted to encode a protein of 735 amino acids with 82% identity to the porcine 17 beta-hydroxysteroid dehydrogenase type IV (HSD IV) originally isolated as a 17 beta-estradiol dehydrogenase. The rat HSD IV was localized to peroxisomes and was regulated by diverse PPC by two distinct mechanisms. Induction of HSD IV and acyl-CoA oxidase (ACO) proteins in rat liver at different treatment times and concentrations of gemfibrozil (GEM) and di-n-butyl phthalate (DBP) were almost identical, suggesting that HSD IV mRNA induction involves the peroxisome proliferator activated receptor alpha, a regulator of ACO. In contrast, HSD IV protein levels were only weakly induced by WY, a strong inducer of ACO protein, even though the levels of both HSD IV and ACO mRNA were strongly stimulated by WY. Thus HSD IV protein levels were uniquely regulated pretranslationally by WY. In addition to HSD IV we also identified the male-specific alpha 2u-globulin as a PPC down regulated gene. This prompted us to examine the expression of another male specific gene, CYP2C11, that catalyzes the hydroxylations of estradiol at the 2 and 16 alpha positions. Cyp2C11 protein expression in rat liver was either decreased or completely abolished after a 3-week treatment by GEM or WY, respectively. Decreased expression of enzymes which inactivate estradiol including Cyp2C11, and the reported increased expression of aromatase may explain why male rats exposed to diverse PPC have higher serum estradiol levels. These higher estradiol levels in male rats have been thought to be mechanistically linked to Leydig cell hyperplasia and adenomas. Increased conversion of estradiol to the less active estrone by HSD IV induction may explain how exposure to the phthalate di-(2-ethylhexyl) phthalate leads to decreases in serum estradiol levels and suppression of ovulation in female rats. PMID- 9209715 TI - Bioelectric potential gradients may initiate cell cycling: ELF and zeta potential gradients may mimic this effect. AB - When a number of experimental studies in bioelectromagnetics were reviewed, those in which weak, exogenous extremely low frequency (ELF) fields were applied in fixed juxtaposition to their target tissues, were found to initiate mitogenesis or mitogenesis-related signals more successfully than when the target tissue moved freely during the irradiation. It is suggested that ELF fields in fixed juxtaposition to their target tissue and implanted foreign bodies or endogenous tissues with a significant zeta potential, mimic bioelectric fields generated at wounds. When the potential is high enough, they assist healing by moving cells into the wound and stimulating quiescent cells at the wound margin to cycle. Electrophoresis may help the initial migration of cells into the wound to form a clot, and migration of fibroblasts and epithelial cells from the wound margin. When exposed for a long time in a fixed juxtaposition to a potential gradient too weak to show in situ microelectrophoresis along the cell membrane surface, surface particles may coalesce to form microclusters, where like-charged surface particles are in close proximity, and growth factor receptor oligomerization and other cycle-initiating reactions are facilitated. PMID- 9209716 TI - Interactions of radiofrequency radiation-induced hyperthermia and 2 methoxyethanol teratogenicity in rats. AB - Radiofrequency (RF) radiation is used in a variety of workplaces. In addition to RF radiation, many workers are concurrently exposed to numerous chemicals; exposed workers include those involved with the microelectronics industry, plastic sealers, and electrosurgical units. The developmental toxicity of RF radiation is associated with the degree and duration of hyperthermia induced by the exposure. Previous animal research indicates that hyperthermia induced by an elevation in ambient temperature can potentiate the toxicity and teratogenicity of some chemical agents. We previously demonstrated that combined exposure to RF radiation (10 MHz) and the industrial solvent, 2-methoxyethanol (2ME), produces enhanced teratogenicity in rats. The purpose of the present research is to determine the effects of varying the degree and duration of hyperthermia induced by RF radiation (sufficient to maintain colonic temperatures at control [38.5], 39.0, 40.0, or 41.0 degrees C for up to 6 h) and 2ME (100 mg/kg) administered on gestation day 13 of rats. Focusing on characterizing the dose-response pattern of interactions, this research seeks to determine the lowest interactive effect level. Day 20 fetuses were examined for external and skeletal malformations. The results are consistent with previous observations. Significant interactions were observed between 2ME and RF radiation sufficient to maintain colonic temperatures at 41 degrees C for 1 h, but no consistent interactions were seen at lower temperatures even with longer durations. These data indicate that combined exposure effects should be considered when developing both RF radiation and chemical exposure guidelines and intervention strategies. PMID- 9209717 TI - The effects of 50 Hz magnetic field exposure on dimethylbenz(alpha)anthracene induced thymic lymphoma/leukemia in mice. AB - Several epidemiological investigations have suggested an increased incidence of lymphoma, leukemia, and brain tumor in residents living near power transmission lines. However, some observers failed to confirm such a positive correlation. To evaluate the effects of extremely low frequency (ELF) magnetic fields on leukemogenesis, an experimental animal model was used, in which thymic lymphoma/leukemia was induced by dimethylbenz(alpha)anthracene (DMBA) injected subcutaneously into the interscapular region of newborn mice within 24 h after birth. Beginning at the second week of age, 165 mice were exposed to 50 Hz magnetic field at 1 mT, 3 h/day, 6 days/week for 16 weeks, and 155 animals exposed to sham conditions. All surviving animals were killed by cervical dislocation at the age of 32 weeks and were examined pathologically. The results showed that the incidences of advanced thymic lymphoma, complicated with lymphomatous leukemia, were 21.8 and 23.9% in the two groups, respectively, without statistically significant differences. But dense metastatic infiltration by lymphoma cells into liver in the field exposure group greater (50%) than that in the sham-exposure group (16.2%) was observed (chi 2 = 9.847, P < 0.01). To determine whether ELF acts as a tumor promoter, further experiments are required. PMID- 9209718 TI - Effects function analysis of ELF magnetic field exposure in the electric utility work environment. AB - The incomplete understanding of the relation between power-frequency fields and biological responses raises problems in defining an appropriate metric for exposure assessment and epidemiological studies. Based on evidence from biological experiments, one can define alternative metrics or effects functions that embody the relationship between field exposure patterns and hypothetical health effects. In this paper, we explore the application of the "effects function" approach to occupational exposure data. Our analysis provides examples of exposure assessments based on a range of plausible effects functions. An EMDEX time series data set of ELF frequency (40-800 Hz) magnetic field exposure measurements for electric utility workers was analyzed with several statistical measures and effects functions: average field strength, combination of threshold and exposure duration, and field strength changes. Results were compared for eight job categories: electrician, substation operator, machinist, welder, plant operator, lineman/splicer, meter reader, and clerical. Average field strength yields a different ranking for these job categories than the ranks obtained using other biologically plausible effects functions. Whereas the group of electricians has the highest exposure by average field strength, the group of substation operators has the highest ranking for most of the other effects functions. Plant operators rank highest in the total number of field strength changes greater than 1 microT per hour. The clerical group remains at the lowest end for all of these effects functions. Our analysis suggests that, although average field strength could be used as a surrogate of field exposure for simply classifying exposure into "low" and "high," this summary measure may be misleading in the relative ranking of job categories in which workers are in "high" fields. These results indicate the relevance of metrics other than average field strength in occupational exposure assessment and in the design and analysis of epidemiological studies. PMID- 9209719 TI - No short-term effects of high-frequency electromagnetic fields on the mammalian pineal gland. AB - There is ample experimental evidence that changes of earth-strength static magnetic fields, pulsed magnetic fields, or alternating electric fields (60 Hz) depress the nocturnally enhanced melatonin synthesis of the pineal gland of certain mammals. No data on the effects of high-frequency electromagnetic fields on melatonin synthesis is available. In the present study, exposure to 900 MHz electromagnetic fields [0.1 to 0.6 mW/cm2, approximately 0.06 to 0.36 W/kg specific absorption rate (SAR) in rats and 0.04 W/kg in Djungarian hamsters; both continuous and/or pulsed at 217 Hz, for 15 min to 6 h] at day or night had no notable short-term effect on pineal melatonin synthesis in male and female Sprague-Dawley rats and Djungarian hamsters. Pineal synaptic ribbon profile numbers (studied in rats only) were likewise not affected. The 900 MHz electromagnetic fields, unpulsed or pulsed at 217 Hz, as applied in the present study, have no short-term effect on the mammalian pineal gland. PMID- 9209720 TI - The role of temporal sensing in bioelectromagnetic effects. AB - Experiments were conducted to see whether the cellular response to electromagnetic (EM) fields occurs through a detection process involving temporal sensing. L929 cells were exposed to 60 Hz magnetic fields and the enhancement of ornithine decarboxylase (ODC) activity was measured to determine cellular response to the field. In one set of experiments, the field was turned alternately off and on at intervals of 0.1 to 50 s. For these experiments, field coherence was maintained by eliminating the insertion of random time intervals upon switching. Intervals < or = 1 s produced no enhancement of ODC activity, but fields switched at intervals > or 10 s showed ODC activities that were enhanced by a factor of approximately 1.7. These data indicate that it is the interval over which field parameters (e.g., amplitude or frequency) remain constant, rather than the interval over which the field is coherent, that is critical to cellular response to an EMF. In a second set of experiments, designed to determine how long it would take for cells to detect a change in field parameters, the field was interrupted for brief intervals (25-200 ms) once each second throughout exposure. In this situation, the extent of EMF-induced ODC activity depended upon the duration of the interruption. Interruptions > or = 100 ms were detected by the cell as shown by elimination of field-induced enhancement of ODC. That two time constants (0.1 and 10 s) are involved in cellular EMF detection is consistent with the temporal sensing process associated with bacterial chemotaxis. By analogy with bacterial temporal sensing, cells would continuously sample and average an EM field over intervals of about 0.1 s (the "averaging" time), storing the averaged value in memory. The cell would compare the stored value with the current average, and respond to the EM field only when field parameters remain constant over intervals of approximately 10 s (the "memory" time). PMID- 9209721 TI - Detection of magnetism in the red imported fire ant (Solenopsis invicta) using magnetic resonance imaging. AB - Red imported fire ant (Solenopsis invicta) workers, queens, and alates were analyzed by magnetic resonance imaging (MRI) for the presence of natural magnetism. Images of ants showed distortion patterns similar to those of honey bees and monarch butterflies, both of which possess ferromagnetic material. The bipolar ring patterns of MRI indicated the presence in fire ants of small amounts of internal magnetic material, which may be used in orientation behaviors, as in the honey bees. PMID- 9209722 TI - Comment on "Effects of 60 Hz electromagnetic fields on early growth in three plant species and a replication of previous results" by Mark S. Davies. PMID- 9209723 TI - A quantitative magnetic resonance imaging study of cerebral and cerebellar gray matter volume in primary unipolar major depression: relationship to treatment response and clinical severity. AB - The authors investigated whether there were differences in cerebral and cerebellar gray and white matter volumes in depressed patients compared to controls, and whether this was associated with treatment response to fluoxetine. Brain magnetic resonance images were obtained from 38 unipolar depressed patients and 20 age, gender, and educationally matched comparison subjects. Patients were divided into groups of "responders" and "nonresponders" based on change in 17 item Hamilton Depression Rating Scale (HDRS) after an 8-week standardized trial of fluoxetine, 20 mg/day. There were no group mean differences in cerebral or cerebellar tissue volumes between patients and controls, or responders and nonresponders. For nonresponders to fluoxetine treatment, cerebral and cerebellar gray matter volume, and total cerebellar tissue volume decreased as baseline HDRS increased. The results suggest an association between gray matter volume and severity of illness in nonresponders to fluoxetine treatment. PMID- 9209724 TI - Pituitary-adrenal cortical axis measures as predictors of sustained remission in major depression. AB - To determine the relationship of pretreatment hypothalamic-pituitary-adrenal cortical (HPA) axis measures in major depressives to the occurrence of relapse following discontinuation of successful treatment, we compared pretreatment demographic, clinical, and HPA axis measures in 35 patients with DSM-III-R primary major depression divided into two groups. One group (n = 26) required continuing treatment to hold their symptoms in abeyance, and the other group (n = 9) had been successfully tapered off medication, remained in remission, and had been medication-free for at least 1 month. The major features that differentiated the 26 patients who required continuing medication to abate their symptoms from the 9 patients who were successfully discontinued from treatment were trends toward a longer duration of episode prior to initial study and increased baseline corticotropin (ACTH) 1-39, and significantly higher baseline cortisol and cortisol response to ACTH 1-24, in the former group. These results suggest that measures of HPA axis hyperactivity, along with longer duration of the index depressive episode, may predict the need for continuing medication for patients to remain in remission. PMID- 9209725 TI - Panic disorder with familial bipolar disorder. AB - If bipolar disorder is genetically heterogeneous, it may be possible to discern clinically heterogeneous familial subtypes based on differential risk for psychiatric comorbidity, for example panic disorder. We evaluated 528 members of 57 families ascertained for a genetic linkage study of bipolar disorder. Families were assorted according to the panic disorder diagnosis of the bipolar proband; the rates of panic and other disorders in relatives were compared. Eighty-eight percent of the 41 subjects with panic disorder had bipolar disorder. Panic disorder was diagnosed in 18% of family members with bipolar disorder. Ten of 57 bipolar probands had panic disorder. Their bipolar first-degree relatives had a significantly higher prevalence of panic disorder, bipolar II, cyclothymia, and dysthymia, but had lower prevalence of substance abuse than the relatives of the bipolar probands without panic disorder. These findings suggest the testable hypothesis that comorbid panic disorder is a marker of genetic heterogeneity in bipolar disorder. PMID- 9209726 TI - A brain imaging (single photon emission computerized tomography) study of semantic and affective processing in psychopaths. AB - Psychopaths have been described as human predators who use charm, intimidation, and violence to control others and to satisfy their own needs. Underlying their propensity to violate social norms and expectations is a profound lack of empathy, guilt, or remorse, affective processes that have long resisted scientific investigation. Using brain imaging technology we found that psychopaths differed from nonpsychopaths in the pattern of relative cerebral blood flow during processing of emotional words. The results were consistent with the hypothesis that there are anomalies in the way psychopaths process semantic and affective information. PMID- 9209727 TI - Sex differences in olfactory identification and Wisconsin Card Sorting performance in schizophrenia: relationship to attention and verbal ability. AB - We investigated the hypothesis that different prefrontal brain systems (i.e., dorsal vs. ventral) and sex contribute differentially to cognitive deficit in schizophrenia. Performance was assessed among clinically stable, chronic schizophrenic outpatients and matched normal control subjects on olfactory identification [on the University of Pennsylvania Smell Identification Test (UPSIT)] and on executive functions [using the Wisconsin Card Sorting Test (WCST)]. Patients were impaired on both tests compared to controls, and male schizophrenics were impaired on the WCST compared to female schizophrenics. The pattern of results suggests that gender differences on the UPSIT are mildly accentuated in schizophrenia. The data support our previous study indicating that UPSIT performance is largely independent of the executive or attentional deficits typically associated with schizophrenia, with the exception of verbal ability. Further research with larger samples is required to test the hypothesis that there is a severely impaired subgroup of male patients with diffuse prefrontal dysfunctions. PMID- 9209729 TI - Seasonal variation in core temperature regulation during sleep in patients with winter seasonal affective disorder. AB - Nocturnal core temperature during sleep is elevated during depression compared with remission in nonseasonally depressed patients. Similarly, nocturnal core temperature is higher during winter depression compared with remission induced by light treatment in seasonal affective disorder (SAD) patients. We investigated whether nocturnal core temperature in SAD patients naturally becomes lower in summer (during remission) compared with winter (during depression). Twenty-four hour core temperature profiles were obtained in winter and summer in 22 SAD patients and 22 controls. The nocturnal core temperature minima were lower in summer compared with winter in SAD patients (p < .005), but not controls (p > .4). The seasonal changes in nocturnal core temperatures in SAD patients may reflect a unique physiological responsiveness of SAD patients to the change of seasons, and may be intimately related to the seasonal disturbances of mood and energy that are characteristic of SAD. PMID- 9209728 TI - Evaluation of cranial electrostimulation therapy on short-term smoking cessation. AB - The effects of cranial electrical stimulation (CES) on short-term smoking cessation were evaluated in a double-blind study of cigarette smokers who wished to stop smoking. Subjects were randomly assigned to a CES- (n = 51) or a sham treated group (n = 50). On 5 consecutive days subjects received CES treatments (30-microA, 2-msec, 10-Hz pulsed signal) or no electrical current (sham). There were no significant differences between groups on daily cigarettes smoked, exhaled carbon monoxide, urinary cotinine levels, treatment retention, smoking urges, or total tobacco withdrawal scores, although subjects in the CES group had less cigarette craving and anxiety during the first 2 experimental days. The ineffectiveness of CES to reduce withdrawal symptoms and facilitate smoking cessation are similar to results of other clinical studies of CES in drug dependence, although positive effects of CES in animal studies have been reported. PMID- 9209730 TI - Clozapine-induced electroencephalogram changes as a function of clozapine serum levels. AB - Specific electroencephalogram (EEG) changes during clozapine therapy were prospectively studied in a cohort of 50 chronic state hospital patients with schizophrenia who were randomly assigned to one of three nonoverlapping clozapine serum level ranges (50-150 ng/mL, 200-300 ng/mL, and 350-450 ng/mL). EEGs were obtained before clozapine was instituted, and after 10 weeks of treatment. Fifty three percent of patients showed EEG changes during the 10-week study period. We observed three seizures (6%), one in a patient on 900 mg (serum level 320 ng/mL) clozapine, and two in patients with lower clozapine serum levels (200-300 ng/mL) who had prior histories of seizures and inadequate valproate coverage. Thirteen percent of patients developed spikes with no relationship to dose or serum level of clozapine. Fifty-three percent of patients developed slowing on EEG. Compared to plasma levels below 300 ng/mL, a clozapine serum level between 350 and 450 ng/mL led to more frequent and more severe slowing. The EEG slowing correlated with observed sleepiness, although this factor was not sufficient to explain the severity of high-dose effects. PMID- 9209731 TI - Limitations of controlled augmentation trials in schizophrenia. AB - To assess the empirical basis for add-on augmentation treatments in schizophrenia, this study examined the experimental design components of pharmacologic augmentation trials in schizophrenia and compared them to conventional requirements. A search covering a 5-year period (1988-1992) identified 13 double-blind, placebo-controlled, parallel, add-on neuroleptic augmentation drug trials. The mean number of subjects per trial was 34.5, and the mean number of outcome measures examined was 25.0. The probability for a significant finding by chance was 63%. Mean effect size required to achieve conventional statistical power was 1.6. Mean statistical power (for effect sizes of .5-1.0) was .1-.4. The mean number of subjects actually required for power of .80 was 58-216. The majority of the 13 trials included too few patients and employed too many outcome measures to conclusively prove or disprove therapeutic efficacy. Conclusions drawn from such trials with less than 40-100 subjects or more than one hypothesis must remain tentative at best. PMID- 9209732 TI - Clozapine treatment of persistent paroxysmal dyskinesia associated with concomitant paroxetine and sumatriptan use. PMID- 9209733 TI - In vivo proton magnetic resonance spectroscopy of Alzheimer's disease in the parietal and temporal lobes. PMID- 9209734 TI - Cholecystokinin tetrapeptide-induced calcium mobilization in T cells of patients with panic disorder, major depression, or schizophrenia. PMID- 9209735 TI - Graft-versus-host disease: the viewpoint from the donor T cell. PMID- 9209736 TI - CD6+ T cell depleted allogeneic bone marrow transplantation from genotypically HLA nonidentical related donors. AB - The widespread use of allogeneic bone marrow transplantation (BMT) is limited by the availability of suitable donors. Recent attempts to expand the donor pool by employing HLA matched unrelated marrow have been partially successful. However, severe graft-versus-host disease (GVHD) and graft failure remain obstacles and contribute to the substantial morbidity and mortality associated with matched unrelated BMT. The use of genotypically nonidentical related or unrelated donor marrow could have wider application if problems associated with GVHD could be overcome. Based upon the low incidence of GVHD in recipients of HLA-matched related donor marrow depleted of T cells with T12, an anti-CD6 monoclonal antibody, we applied this approach to 27 adult recipients of HLA mismatched related bone marrow. Ten patients received marrow mismatched at 2 HLA loci, 13 received 1 antigen mismatched marrow, and 4 received phenotypically identical marrow from a non-sibling. Immediately prior to admission, patients were treated with total lymphoid irradiation (750-1050 cGy) to suppress host derived. T lymphocytes capable of mediating graft rejection. The ablative regimen consisted of cyclophosphamide (60 mg/kg x 2 days) followed by total body irradiation (1400 cGy in 7 fractions over 4 days). Patients then received marrow depleted of T cells with T12 (CD6) plus complement. No immune suppressive medications were administered to prevent GVHD. Twenty-four of 27 patients displayed stable hematologic engraftment, achieving an absolute neutrophil count of 0.5 x 10(9)/L at a median of 19 days post-BMT. Degree of HLA disparity did not influence engraftment. Among engrafting patients, grades 2-4 acute GVHD occurred in 40% and grade 3-4 GVHD in 8%. Chronic GVHD developed in 5 patients. Patients mismatched at 2 loci were more likely to develop GVHD than those mismatched at 0-1 loci (logrank, p = .04). Disease relapse has occurred in only 3 patients receiving mismatched marrow. Estimated overall survival for mismatched patients is 56% at 2 years and is independent of HLA disparity. Among the patients transplanted for chronic myelogenous in stable phase or acute leukemia in first remission, estimated event free survival is 69% at 2 years compared to 20% for patients with more advanced disease. Our results suggest that transplantation of mismatched related marrow using modalities designed to reduce GVHD without immune suppressive medication (CD6 depletion) is feasible and should prompt wider investigation into the extended families of patients in the search for potential marrow donors. This approach also merits investigation in recipients of matched unrelated marrow as a potential means of reducing transplant-related toxicity. PMID- 9209737 TI - Comparison of long-term outcome of children with severe aplastic anemia treated with immunosuppression versus bone marrow transplantation. AB - Children with severe aplastic anemia (SAA) are treated with bone marrow transplantation (BMT) if a human leukocyte antigen (HLA) compatible sibling donor is available, or alternatively with immunosuppressive therapy (IST). Three retrospective trials examining BMT vs IST in pediatric patients treated from 1970 1988 found BMT resulted in a superior survival rate. Advances have been made in general supportive care and in the approach to each of these treatment modalities in the last decade. To compare survival following BMT and IST in a more recent era, we retrospectively analyzed the results of 48 consecutively treated children with SAA presenting to Memorial Sloan-Kettering Cancer Center (MSKCC) between 1983 and 1992. In contrast to the previous studies, the estimated survival of the BMT and IST groups at 120 months are equivalent, 75.6% and 73.8%, respectively. The IST results in our series are superior to the 42-48% (2-10 year) survival previously published, but similar to survival data observed in more recent IST trials employing more intensive immunosuppression (antithymocyte globulin and cyclosporine). The overall BMT survival rates are similar to those previously published, although BMT results improved dramatically during the latter five years of this analysis, with all 11 patients transplanted surviving with a minimum follow-up of 3 years. No surviving BMT patient has extensive chronic graft-versus-host disease (GvHD), a major cause of long-term mortality post-BMT. Therefore, it is likely the BMT survival curve will remain stable. In contrast, the survival curve of the IST patients is likely unstable, since patients are still at risk for relapse or development of clonal disease. Thus, despite overall similar survival rates, we continue to recommend BMT as first-line therapy in pediatric SAA patients with matched sibling donors. PMID- 9209738 TI - The outcome of unrelated donor bone marrow transplantation in patients with hematologic malignancies using tacrolimus (FK506) and low dose methotrexate for graft-versus-host disease prophylaxis. AB - Initial studies of FK506 combined with methotrexate (MTX) in patients receiving unrelated donor BMT have demonstrated a possible-decrease in the incidence of severe GVHD but high rates of severe stomatitis and nephrotoxicity. With this background, we undertook a pilot study evaluating FK506 in combination with a lower than usual dose of MTX in an attempt to improve the tolerability of this immunoprophylaxis regimen. Between July 1993 and October 1994, 26 consecutive adults receiving unrelated donor BMT at Emory University Hospital were enrolled on this study. All patients received FK506 intravenously at an initial dose of 0.03 mg/kg/day beginning day -1 and continuing until oral FK506 was tolerated. Patients also received MTX intravenously at 5 mg/m2 on days 1, 3, 6, and 11. The preparative regimen administered to all but one patient included cyclophosphamide at 200 mg/kg over 4 days followed by total body irradiation (TBI) at 1400 cGy in twice daily fractions over 4 days. The median age of patients was 31 years (range: 19 to 52). Sixteen donor/recipient pairs were matched for HLA-A, -B, and DR by serology and molecular typing. Ten paris were minor mismatches at either class I or class II. Twenty-two of 26 patients (85%) completed four doses of MTX on schedule. Nephrotoxicity was the most common adverse event associated with the administration of FK506: 88% of patients experienced a doubling of their serum creatinine. One patient died of central nervous system hemorrhage prior to engraftment. Twenty-four of the remaining 25 patients (96%) engrafted. Fourteen of 24 patients (50%) evaluable developed grades 2-4 acute GVHD. The rate of severe (grades 3-4) acute GVHD was 25%. Chronic GVHD developed in 11 of 20 (55%) evaluable patients. At a median follow-up of 461 days, 14 patients (54%) are alive. All are relapse-free with a median Karnofsky performance status of 90% (range: 70-100%). The cumulative probability of 2-year disease-free survival is 50% (95% confidence interval [CI]: 0.33 to 0.77); for low risk patients 67% (95% CI: 0.47 to 0.95) and for high risk patients 23% (95% CI: 0.049 to 1.00). These preliminary data indicate that FK506-based immunosuppression following unrelated donor BMT is effective in preventing severe acute GVHD and warrants comparison to CSA-based regimens. PMID- 9209739 TI - Low dose subcutaneous interleukin-2 after autologous transplantation generates sustained in vivo natural killer cell activity. AB - Autologous transplantation can induce extended remission in some patients with advanced breast cancer and lymphoma yet nearly 80% and 50%, respectively, will ultimately relapse. In vitro studies suggest that activated natural killer cells (NK) mediate lytic activity against breast cancer and lymphoma cell lines. Therefore, immunotherapy with interleukin-2 (IL-2, Amgen) to activate NK may improve long-term disease-free survival when administered in a post-transplant minimal residual disease setting. To determine the feasibility of administering IL-2 and activation of NK post-transplant, twelve patients (6 breast cancer, 6 lymphoma) were enrolled on a phase I dose escalation study after autologous transplantation (median day + 94, range 50-166). IL-2 was self administered at 0.25 x 10(6) (n = 6) or 0.5 x 10(6) (n = 6) U/m2/day subcutaneously for 84 consecutive days. The best tolerated dose was 0.25 x 10(6) U/m2/day (75% of planned doses given vs. 48% at the higher dose). Dose limiting toxicity occurred in 6 patients (n = 2 at 0.25 x 10(6) U/m2/day, n = 4 at 0.5 x 10(6) U/m2/day) consisting of decreased performance status (n = 2), thrombocytopenia (n = 3, 1 at the lower dose), and mild neutropenia (n = 1 at the lower dose). However, all symptoms resolved within a week following discontinuation of IL-2 and no patient required hospitalization. Circulating soluble IL-2 receptor levels were significantly increased in all patients receiving IL-2. Patients receiving at least 28 days of IL-2 exhibited a greater than 10-fold increment in circulating CD56+bright/CD3- NK. Furthermore, lytic function was increased against NK resistant targets, MCF-7 (breast cancer), and Raji (lymphoma). In vivo IL-2 primed NK cells obtained by lymphapheresis were activated in large-scale ex vivo incubation in high dose IL-2 (1,000 U/mL) at high cell density (10 x 10(6)/mL), in gas permeable bags, and using serum-free media. NK lytic function against MCF 7 and Raji targets was further enhanced. We conclude that low dose subcutaneous IL-2 based immunotherapy is feasible, relatively safe, can be administered in an outpatient setting and hypothesize that additional ex vivo incubation in IL-2 may be used to generate NK cells with potent antitumor effects in vivo. PMID- 9209741 TI - Surfactant therapy in the USA: trials and current routines. PMID- 9209740 TI - Spontaneous splenic rupture following administration of granulocyte colony stimulating factor (G-CSF): occurrence in an allogeneic donor of peripheral blood stem cells. AB - Granulocyte colony-stimulating factor (G-CSF) has been used to improve granulocyte count in chronic neutropenia and myelodysplasia, to minimize the incidence and duration of neutropenia during conventional chemotherapy, and to mobilize peripheral blood stem cells prior to leukapheresis for use in autologous and allogeneic marrow transplantation. The most common toxicity is bone pain, and other reactions such as inflammation at the site of injection have also occurred. In patients with chronic neutropenia, splenomegaly has been described with long term use, and extramedullary hematopoiesis has also been reported. However, thus far, no life-threatening sequelae of these effects are found in the literature. We now describe a case of spontaneous splenic rupture four days following a six day course of G-CSF therapy in an allogeneic donor of peripheral blood stem cells. PMID- 9209742 TI - Clinical trials of surfactant replacement in Europe. PMID- 9209743 TI - Ventilation patterns, surfactant and lung injury. PMID- 9209744 TI - Circulatory effects of surfactant therapy. PMID- 9209746 TI - From bubbles to babies: the evolution of surfactant replacement therapy. PMID- 9209745 TI - Free oxygen radicals and surfactant. PMID- 9209747 TI - Resuscitation strategy and surfactant therapy. PMID- 9209749 TI - Clinical aspects of nitric oxide and surfactant replacement. PMID- 9209748 TI - Experimental models of bronchopulmonary dysplasia. PMID- 9209750 TI - Molecular interactions between nitric oxide and lung surfactant. PMID- 9209751 TI - Protein analogues in artificial surfactants. PMID- 9209752 TI - Artificial life: a bridge toward a new artificial intelligence. PMID- 9209753 TI - Emergent models of supple dynamics in life and mind. AB - The dynamical patterns in mental phenomena have a characteristic suppleness--a looseness or softness that persistently resists precise formulation--which apparently underlies the frame problem of artificial intelligence. This suppleness also undermines contemporary philosophical functionalist attempts to define mental capacities. Living systems display an analogous from of supple dynamics. However, the supple dynamics of living systems have been captured in recent artificial life models, due to the emergent architecture of those models. This suggests that analogous emergent models might be able to explain supple dynamics of mental phenomena. These emergent models of the supple mind, if successful, would refashion the nature of contemporary functionalism in the philosophy of mind. PMID- 9209754 TI - Computing machinery and mentality. AB - I reconsider the status of computationalism (or, in a weak sense, functionalism): the claim that being a realization of some (as yet unspecified) class of abstract machine is both necessary and sufficient for having genuine, full-blooded, mentality. This doctrine is now quite widely (though by no means universally) seen as discredited. My position is that, though it is undoubtedly an unsatisfactory (perhaps even repugnant) thesis, the arguments against it are still rather weak. In particular, I critically reassess John Searle's infamous Chinese Room Argument and also some relevant aspects of Karl Popper's theory of the Open Universe. I conclude that the status of computationalism must still be regarded as undecided, and that it may still provide a satisfactory framework for research. PMID- 9209755 TI - Symbol grounding: a bridge from artificial life, to artificial intelligence. AB - This paper develops a bridge from AL issues about the symbol-matter relation to AI issues about symbol-grounding by focusing on the concepts of formality and syntactic interpretability. Using the DNA triplet-amino acid specification relation as a paradigm, it is argued that syntactic properties can be grounded as high-level features of the non-syntactic interactions in a physical dynamical system. This argument provides the basis for a rebuttal of John Searle's recent assertion that syntax is observer-relative (1990, 1992). But the argument as developed also challenges the classic symbol-processing theory of mind against which Searle is arguing, as well as the strong AL thesis that life is realizable in a purely computational medium. Finally, it provides a new line of support for the autonomous systems approach in AL and AI (Varela & Bourgine 1992a, 1992b). PMID- 9209756 TI - Patterns of life: intertwining identity and cognition. PMID- 9209757 TI - Autonomy in robots and other agents. AB - The word "autonomous" has become widely used in artificial intelligence, robotics, and, more recently, artificial life and is typically used to qualify types of systems, agents, or robots: we see terms like "autonomous systems," "autonomous agents," and "autonomous robots." Its use in these fields is, however, both weak, with no distinctions being made that are not better and more precisely made with other existing terms, and varied, with no single underlying concept being involved. This ill-disciplined usage contrasts strongly with the use of the same term in other fields such as biology, philosophy, ethics, law, and human rights, for example. In all these quite different areas the concept of autonomy is essentially the same, though the language used and the aspects and issues of concern, of course, differ. In all these cases the underlying notion is one of self-law making and the closely related concept of self-identity. In this paper I argue that the loose and varied use of the term autonomous in artificial intelligence, robotics, and artificial life has effectively robbed these fields of an important concept. A concept essentially the same as we find it in biology, philosophy, ethics, and law, and one that is needed to distinguish a particular kind of agent or robot from those developed and built so far. I suggest that robots and other agents will have to be autonomous, i.e., self-law making, not just self-regulating, if they are to be able effectively to deal with the kinds of environments in which we live and work: environments which have significant large scale spatial and temporal invariant structure, but which also have large amounts of local spatial and temporal dynamic variation and unpredictability, and which lead to the frequent occurrence of previously unexperienced situations for the agents that interact with them. PMID- 9209758 TI - Cognition and life: the autonomy of cognition. AB - In this paper we propose a philosophical distinction between biological and cognitive domains based on two conditions that are postulated to obtain a useful characterization of cognition: biological grounding and explanatory sufficiency. According to this, we argue that the origin of cognition in natural systems (cognition as we know it) is the result of the appearance of an autonomous system embedded into another more generic one: the whole organism. This basic idea is complemented by another one: the formation and development of this system, in the course of evolution, cannot be understood but as the outcome of a continuous process of interaction between organisms and environment, between different organisms, and, specially, between the very cognitive organisms. Finally, we address the problem of the generalization of a theory of cognition (cognition as it could be) and conclude that this work would imply a grounding work on the problem of the origins developed in the frame of a confluence between both Artificial Life and an embodied Artificial Intelligence. PMID- 9209760 TI - Artificial life and higher level cognition. PMID- 9209759 TI - Artificial evolution: a new path for artificial intelligence? AB - Recently there have been a number of proposals for the use of artificial evolution as a radically new approach to the development of control systems for autonomous robots. This paper explains the artificial evolution approach, using work at Sussex to illustrate it. The paper revolves around a case study on the concurrent evolution of control networks and visual sensor morphologies for a mobile robot. Wider intellectual issues surrounding the work are discussed, as is the use of more abstract evolutionary simulations as a new potentially useful tool in theoretical biology. PMID- 9209761 TI - Evolution of neural ontogenies: the ontogeny and phylogeny of invertebrate and vertebrate nervous systems. Introduction. PMID- 9209762 TI - The evolution of insect flight: implications for the evolution of the nervous system. AB - The Insecta encompasses a prodigiously diverse group as measured at the species, family and ordinal levels, but the nervous system bears evidence of conservatism. The early acquisition of flight must have been a major factor in the diversification of body form. Arguments are presented that predator evasion was a primary factor in the origin of flight and that a conserved set of giant interneurons played a key element in the transition. PMID- 9209763 TI - The evolution of insect wings and their sensory apparatus. AB - The development of wings has undoubtedly played a major role in the enormous diversification of insects. New insights into the evolutionary history of insect wings are available from paleontological, physiological and biomechanical studies. A recent hypothesis, derived primarily from paleontological evidence, is that wings arose from leg exites, small flaps associated with proximal leg segments. We present data from studies on physical models that are consistent with this hypothesis. The exites would have been moveable, and measurements on scaled models show that they would have generated aerodynamic lift by unsteady mechanisms associated with vortex shedding. An examination of the sensory structures found on insect wings is also consistent with the interpretation of protowings as leg exites. In addition to mechanosensory bristles, such as are found all over the body, the wings of modern insects carry campaniform sensilla sensitive to cuticular deformation and contact chemoreceptors whose stimulation elicits a feeding response. Both classes of receptors are also found on the legs of modern insects but not on the thorax, favoring the leg exite theory. PMID- 9209764 TI - Evolution of gnathostome lateral line ontogenies. AB - An outgroup analysis of multiple ontogenies provides the most robust approach to understanding phylogeny. Such an analysis of the lateral line system among extinct and extant gnathostomes reveals that lateral line placodes constitute the basic ontogenetic unit responsible for the development of this system. Six pairs of lateral line placodes appear to have existed in the earliest gnathostomes, and eight stages (stages A-H) can be recognized in their differentiation. Terminal truncation (heterochronic changes) in the primitive sequence of placodal development has occurred in one or more placodes in each gnathostome radiation, with the most extensive truncations occurring in arthrodire placoderms, lepidosirenid lungfishes and extant amphibians. The most extensive nonterminal changes in the primitive sequence of placodal development involve the failure of electroreceptors to form within the lateral zones of the elongatiang sensory ridges of the placodes. This nonterminal change appears to have occurred independently in ancestral neopterygian bony fishes, in many amphibians and, possibly, in the extinct acanthodians. At least two teleost radiations, osteoglossomorphs and ostariophysines, have re-evolved electroreceptors which may represent additional nonterminal changes in placodal patterning or, possibly, a change in the embryonic source of these receptors. PMID- 9209765 TI - On the role played by ontogenetic remodeling and functional transformation in the evolution of terrestrial hearing. AB - Using examples from the octavolateral system, evidence is reviewed suggesting a relationship between regressive events, such as loss of one function, or loss of one sensory subsystem, and progressive evolutionary changes in topologically associated systems. While none of the neuronal examples in the evolutionary reorganization of the otic region are as clear-cut as the initial example of non neuronal reorganization on which the correlation of regressive with progressive changes is based (the functional transformation of the hyomandibular bone into the stapes), the general principle that a chance correlation of two insignificant events may lead to a novel function may be valid for more aspects of the evolution of the ear, in particular the auditory system, than is currently appreciated. It is suggested that regressive events may not only be an evolutionary dead end but that they may provide, through the relaxation of constraints imposed on the respective structure, a source for innovations. However, transformations of functionally uncoupled structures into a novel adaptive function will occur only when topologically adjacent structures require these transformations to improve their own function. PMID- 9209766 TI - Genome size, secondary simplification, and the evolution of the brain in salamanders. AB - Compared to other vertebrates, even including lampreys and hagfishes in some respects, salamanders exhibit a relatively simple organization of brain and sense organs which is illustrated here using the visual system as an example. The greatest simplicity is found in the bolitoglossine salamanders, yet all bolitoglossines possess highly projectile tongues and rely on vision for survival; furthermore, some species are agile and acrobatic. The unusual features of the visual system of salamanders include small numbers of large neurons, a low degree of morphological differentiation among neurons, a small proportion of myelinated axons in the optic nerve, and an optic tectum consisting essentially of a periventricular cellular layer and a superficial fiber layer. Similar features are found throughout the central nervous system of salamanders and in the lateral line, auditory and olfactory systems as well. Phylogenetic analysis shows that the most parsimonious interpretation of these data is that the simple organization of the brain and sense organs of salamanders was derived secondarily from a more complex ancestral state. We hypothesize that increased genome size has led to simplification of the nervous system in salamanders. Increased genome size appears to have had profound effects on neural development in salamanders, leading to paedomorphosis, the retention of juvenile or even embryonic characteristics into adulthood. In particular, large genome size is associated with large cell size and reduced rates of cell proliferation, migration and differentiation. Secondary simplification has constrained the function of the salamanders' visual system, primarily by increasing cell size and decreasing cell numbers. However, it also has provided an opportunity for the evolution of compensating mechanisms, which have helped to restore or even enhance visual function. Most apparent among the compensatory mechanisms of bolitoglossine salamanders is the presence of well developed ipsilateral retinotectal projections, which apparently enhance depth perception. It is difficult to explain the unusual history of the nervous system in salamanders solely in terms of natural selection and adaptation. Increasing genome size through selfish replication appears to have played a major role in the evolution of salamander brains by imposing functional constraints as well as creating opportunities for overcoming them. PMID- 9209767 TI - A 40-year follow-up of school children with migraine. AB - A prevalence study of 9000 Swedish school children conducted in 1955 showed that nearly 4% had migraine. The prevalence of migraine was 1.4% at 7 years of age and 5.3% at 15 years of age. From the age of 11 there was a gradual increase of migraine headache and a predominance among girls. A subgroup of 73 children with pronounced migraine and an average onset of 6 years was followed during a period of 40 years. The results showed that 23% of the children were migraine-free before the age of 25, boys significantly more often than girls. However, around the age of 50, more than half of the migraine group still had migraine attacks. A recall bias was found in that a number of the subjects in their middle-life (41%) could not remember that they had had aura symptoms previously. Of those who had become parents, 52% have in their present or previous families had one child or more who had developed recurrent headache, probably of the migraine-type. PMID- 9209768 TI - Prevalence of sleep disorders in childhood and adolescence with headache: a case control study. AB - Although a relationship between headache and sleep disturbances has been reported in adults, only few data have been available in children. Accordingly, we performed a survey to determine the prevalence of sleep disturbances in children with migraine and tension-type headache. A questionnaire of history and clinical data and of sleep disturbances was given to parents of 283 headache subjects (164 with migraine and 119 with tension-type headache). Results were compared to a normative group comparable for age and sex of 893 normal healthy subjects. Migraine subjects showed a higher prevalence of sleep disturbances during infancy as well as 3-month colic. In both headache groups, more parents had sleep disturbances and there was a higher occurrence of co-sleeping and napping. A high frequency of sleep disturbances involving sleep quality, night awakenings, nocturnal symptoms and daytime sleepiness was reported in headache children. No statistical differences were found in the prevalence of sleep disturbances between migraine and tension-type headache. However, the migraine group tended to have "disturbed sleep" more often with increased prevalence of nocturnal symptoms such as sleep breathing disorders and parasomnias. Our results give further support to an association between sleep and migraine that may have a common intrinsic origin. PMID- 9209769 TI - Symptoms and diseases and smoking habits in female episodic cluster headache and migraine patients. AB - Twenty-seven episodic female cluster headache patients were compared to 27 age matched female migraine patients with regard to occurrence of symptoms and diseases other than headache, and also with regard to tobacco consumption. Some symptoms and diseases were found to occur significantly or almost significantly more often in the cluster headache patients than in the migraine patients; Chronic fatigue (p < 0.01), vertigo (p < 0.05), arthralgia (p < 0.05), back pain (p = 0.05), spontaneous ecchymoses (p = 0.05) and constipation and/or periodic diarrhea (p = 0.09). There were significantly fewer persons who had never smoked in the cluster headache group than in the migraine group (p < 0.01). The extent of smoking was significantly greater in the cluster headache group than in the migraine group, both as to the number of cigarettes smoked per day (p < 0.001) and as to smoking years (p < 0.001). PMID- 9209770 TI - Phantom eye: features and prevalence. The predisposing role of headache. AB - We prospectively evaluated the frequency, time-course and predisposing factors of phantom eye syndrome in 53 patients who underwent surgical eye amputation to cure ocular cancer. Before surgery, patients were classified as Group I (n = 25) if they had no history of headache or Group II (n = 28) if they were headache sufferers. Three clinical patterns were distinguished: phantom pain, non-painful phantom phenomena and photopsias. Their symptoms developed 7 days to 6 months after surgery, with peak incidence after 6 months (photopsia 43%; phantom pain 28%; non-painful phantom phenomena 62%). Phantom eye syndrome was more common in headache sufferers than in non-headache subjects. Headache sufferers were more prone to phantom pain, but more so to non-painful phenomena and photopsias. These findings are in accord with our previous results indicating that primary headache sufferers are prone to phantom tooth pain. PMID- 9209771 TI - Migraine-associated vertigo. AB - A retrospective analysis was performed on a consecutive series of 363 patients presenting with vertigo; 32% had migraine. Of the 224 patients with no pathology other than migraine or vestibular dysfunction, migraineurs had a significantly higher prevalence of normal, central, and combined central and peripheral vestibular dysfunction compared to non-migraineurs. The combination of central and peripheral vestibular signs was a feature of migraine with aura. The results support the hypothesis that migraine-associated vertigo is a diagnostic entity. PMID- 9209772 TI - Nocturnal plasma melatonin profile and melatonin kinetics during infusion in status migrainosus. AB - The plasma melatonin profile was significantly disturbed (phase-shift of the maximum melatonin level) in four out of six female sufferers from status migrainosus, compared with nine healthy controls. The number of secretion peaks was similar in both groups. A nocturnal 20 micrograms melatonin infusion (from 21.00 to 01.00 h) evoked plasma melatonin levels slightly higher than a physiological secretion peak. During infusion, the episodes of secretion were reinforced and the endogenous plasma profile was phase-advanced in two patients displaying a phase-delay. These data suggest impaired pineal function in migraine. In the absence of side effects of melatonin infusion, the relief of certain migraine symptoms described by our patients might support a controlled trial of melatonin in migraine. PMID- 9209773 TI - Intravital microscope studies on the effects of neurokinin agonists and calcitonin gene-related peptide on dural vessel diameter in the anaesthetized rat. AB - This study describes a novel intravital microscope technique for direct measurement of dural blood vessel diameter through a closed cranial window in anaesthetized rats. This technique avoids removal of the skull, which can lead to problems of altered vessel reactivity and brain swelling that are encountered with open cranial window techniques. Substance P and calcitonin gene-related (CGRP) evoked increases in dural vessel diameter, which were abolished by the NK1 receptor antagonist, RP67580 and the CGRP receptor antagonist, human-alpha CGRP(8 37) respectively. Neurokinin A produced increases in dural vessel diameter which were unaffected by the NK2 receptor antagonist SR 48968 but were blocked by RP67580, suggesting that neurokinin A can act through NK1 receptors to produce dural vasodilation in rats. The NK3 receptor agonist, senktide, had no effects on dural vessel diameter. All drugs were administered intravenously. In humans, vasodilation within the meningeal vasculature has been implicated in the pathogenesis of migraine, the present experiments indicate that substance P or neurokinin A (both acting through NK1 receptors) or CGRP may be responsible. PMID- 9209774 TI - Sumatriptan inhibits neurogenic vasodilation of dural blood vessels in the anaesthetized rat--intravital microscope studies. AB - This study used intravital microscopy to measure the diameter of dural arteries in anaesthetized rats. Electrical stimulation of the surface of a closed cranial window produced increases in dural vessel diameter which were blocked by the CGRP receptor antagonist human-alpha CGRP(8-37) but unaffected by the NK1 receptor antagonist RP67580. Sumatriptan (3 and 10 mg kg-1, i.v.) significantly reduced the response to electrical stimulation. In contrast, sumatriptan (3 mg kg-1) had no effects on the response to exogenously administered CGRP. These results indicate that neurokinins play no role in neurogenic vasodilation in this preparation and that neurogenic vasodilation in rat dural vessels is mediated predominantly by CGRP. Furthermore, the data indicate that sumatriptan attenuates neurogenic vasodilation, probably by inhibiting the release of CGRP from perivascular trigeminal nerve endings innervating the dura. These experimental data parallel the clinical findings that CGRP levels are elevated in migraine and normalized, concomitantly with headache relief, by sumatriptan. PMID- 9209775 TI - Safety, tolerability, and pharmacokinetics of sumatriptan suppositories following single and multiple doses in healthy volunteers. AB - A suppository formulation of the 5HT1 agonist sumatriptan could prove an important therapeutic option in migraine patients who dislike or poorly tolerate injectable therapy and where oral tablet administration is unsuitable because of severe migraine-related vomiting. Two independent double-blind, randomized clinical studies were conducted to evaluate the safety, tolerability and pharmacokinetics of sumatriptan suppositories following ascending single doses (four different dose levels) and multiple doses. In the four-period, crossover, single-dose study, 24 healthy male subjects were randomized to receive a suppository containing 12.5, 25, 50, or 100 mg on separate occasions 3-14 days apart. The suppositories were generally well tolerated; transient asthenia, drowsiness, and headache were the most frequently reported adverse events, and these were not dose-related. Peak plasma concentrations (Cmax) of sumatriptan were proportional to dose from 25 to 100 mg; area under the plasma concentration time curve (AUC infinity) values were proportional to dose except at the highest doses, when they were greater than those predicted from lower doses. For all doses, the tmax of sumatriptan occurred within 2.5 h, and the t1/2 was approximately 2 h. In the two-period, placebo-controlled, crossover, repeat-dose study, 12 healthy adult male subjects were randomized to receive either a 50-mg sumatriptan suppository or placebo suppository, administered rectally twice a day, for 11 doses (5 1/2 days). Adverse events were no more frequent with sumatriptan than with placebo, and stool guaiac, rectal examinations, and physical examinations remained normal. No significant differences were noted between Day 1 and Day 6 values in the AUC, Cmax, time of peak serum concentration (tmax), elimination half-life (t 1/2), fraction of the dose excreted in the urine (fe), or renal clearance (Clr) of sumatriptan or its pharmacologically inactive indole acetic acid metabolite. Serum metabolite concentrations were two to three fold higher than corresponding sumatriptan concentrations. No clinically significant accumulation of sumatriptan or its metabolite occurred. Overall, these studies show that sumatriptan administration via a suppository formulation is well tolerated, allows rapid absorption of sumatriptan, results in sumatriptan Cmax values that are proportional to dose from 25 to 100 mg, and is not associated with accumulation of sumatriptan or its metabolite. PMID- 9209776 TI - Safety, tolerability, and pharmacokinetics of sumatriptan in healthy subjects following ascending single intranasal doses and multiple intranasal doses. AB - The delivery of sumatriptan doses intranasally could add greater flexibility in the treatment of migraine than is possible with the currently available subcutaneous and oral sumatriptan preparations. Two independent double-blind, randomized, placebo-controlled clinical studies were conducted to evaluate the safety, tolerability and pharmacokinetics of intranasally administered sumatriptan following ascending single doses (three different dose levels) and multiple doses. In the four-way, crossover, ascending-dose study, 20 healthy female subjects were randomized to receive on separate occasions single intranasal spray doses of 5, 10, or 20 mg sumatriptan (as the hemisulphate salt) or placebo into one nostril. Adverse events were mild and consisted mainly of bitter taste at the back of the throat and events typical of sumatriptan administered by other routes (headache, lightheadedness and tingling). Area under the plasma sumatriptan concentration versus time curve (AUC infinity) and peak plasma concentration (Cmax) increased with the dose. Dose proportionality was demonstrated between 5 and 10 mg but not across the dose range 5-20 mg. Time to maximum plasma concentration (tmax) was variable due to multiple peaking. The elimination half-life (t1/2), approximately 2 h, was unaffected by the magnitude of dose. In the two-period, multiple-dose, crossover study, 12 healthy adult male and female subjects were randomized to receive either sumatriptan hemisulphate 20 mg or placebo, administered intranasally as a spray three times a day for 4 days. The two dosing periods were separated by 3 to 14 days. Multiple doses of sumatriptan were well tolerated, with no serious adverse events occurring or withdrawals due to adverse events. All patients reported a mild to moderate drug related disturbance of taste. Nasal examinations remained normal, and olfactory function was unaffected. The AUC over the first 8 h following dosing (AUC8) and fraction of the dose excreted in the urine (fe; 6.2% vs 3.6%) were similar on Days 1 and 4. Day 4 values were significantly higher (p < or = 0.05) for Cmax (16.9 ng/ml vs 13.1 ng/ml), renal clearance (Clr; 19.0 l/h vs 14.2 l/h), and t1/2 (2.18 h vs 1.93 h), and shorter for tmax (0.88 h vs 1.75 h). Some accumulation (22%) occurred over the 4 days of dosing. Serum concentrations of the pharmacologically inactive indole acetic acid metabolite of sumatriptan were fourfold to fivefold higher than corresponding sumatriptan concentrations. Overall, these studies show that the sumatriptan intranasal spray formulation is well tolerated, allows rapid absorption of sumatriptan, and results in only a clinically insignificant degree of sumatriptan accumulation upon repeated dosing. PMID- 9209777 TI - Transdermal clonidine in the prophylaxis of episodic cluster headache. PMID- 9209778 TI - Serum concentrations and ex vivo inhibitory/bactericidal activity of clindamycin after administration of two oral dosages. AB - Two dosages of clindamycin, 300 and 600 mg, were given orally to 10 patients each. The patients were admitted for minor elective surgery. Their mean age was 39.3 years; mean weight was 67.5 kg. None of them had taken antibiotics for at least 1 month. After administration of a single dose, blood samples were obtained at 0.5, 1, 1.5, 2, 3, 4, 6, 8, 10 and 12 h after dosing. Drug levels were determined by the bioassay method using Micrococcus luteus as test organism. Serum inhibitory and bactericidal activities against five isolates each of Staphylococcus aureus and Streptococcus pyogenes were determined by the microdilution method according to the National Committee for Clinical Laboratory Standards guidelines. The mean peak serum level was 3.4 mg/l for the 300-mg dose and 4.8 mg/l for the 600-mg dose. The mean reciprocal peak inhibitory titers for the 300-/600-mg doses were 13.7/23.8 and 15.2/34.7 against S. aureus and S. pyogenes, respectively. Most serum samples did not show bactericidal activity against S. aureus. PMID- 9209779 TI - Trends of genetic relationship of serotype 23F penicillin-resistant Streptococcus pneumoniae in Japan. AB - No data on the genetic analysis of penicillin-resistant Streptococcus pneumoniae (PRP) in Japan has been reported. The SmaI restriction endonuclease digested patterns of chromosomal DNAs from 15 PRP serotyped 23F isolated in Japan were analyzed by pulsed-field gel electrophoresis (PFGE). The isolates were genetically heterogeneous and seven different PFGE patterns were identified. Nine strains were also resistant to erythromycin and tetracycline. Four strains revealed resistance to ceftriaxone. The PFGE patterns of some strains isolated in Nagasaki University Hospital were identical to each other and closely resembled those of isolates from three different areas of Japan. These results indicate a need for additional studies by PFGE to determine the possibility of clonal spread in Japan. PMID- 9209780 TI - Comparison of in vitro antimicrobial activity of AM-1155 with those of tosufloxacin and fleroxacin against clinical isolates of Neisseria gonorrhoeae harboring quinolone resistance alterations in GyrA and ParC. AB - The in vitro antimicrobial activities of AM-1155, a new fluoroquinolone, tosufloxacin and fleroxacin were tested against 55 clinical isolates of Neisseria gonorrhoeae using the agar dilution method. In our previous study, all the strains had been examined for mutations in the region corresponding to the quinolone-resistance determining region of the Escherichia coli gyrA gene and the analogous region of the parC gene, and tested for susceptibility to ciprofloxacin. In this study, the 55 isolates of N. gonorrhoeae were assigned to one of three categories based on the presence or absence of alterations in GyrA and ParC. In each category, the antimicrobial activity of AM-1155 against the isolates was compared with those of tosufloxacin and fleroxacin. The MICs of AM 1155 for 11 highly fluoroquinolone-resistant isolates with alterations in both GyrA and ParC ranged from 0.06 to 1.0 microgram/ml. The MICs inhibiting 50% (MIC50) and 90% (MIC90) of these isolates were 0.125 and 1.0 microgram/ml, respectively. The MICs of AM-1155 for 20 moderately fluoroquinolone-resistant isolates with alterations only in GyrA ranged from 0.03 to 0.25 microgram/ml (MIC50, 0.06 microgram/ml; MIC90m, 0.125 microgram/ml). The MICs of AM-1155 for 24 of the quinolone-susceptible isolates without alterations in either GyrA or ParC ranged from 0.004 to 0.03 microgram/ml (MIC50, 0.008 microgram/ml. MIC90, 0.015 microgram/ml). There were significant differences between the MIC distribution of AM-1155 and each corresponding MIC distribution of tosufloxacin and fleroxacin in these three categories to which the 55 isolates were assigned (p < 0.05). Based on the MIC90S of the tested fluoroquinolones, AM-1155 was two- and eightfold more active against the highly fluoroquinolone-resistant isolates than tosufloxacin and fleroxacin, respectively. Against the moderately fluoroquinolone-resistant isolates, AM-1155 was four- and sixteenfold more active than tosufloxacin and fleroxacin, respectively. Against the quinolone-susceptible strains, AM-1155 was also two- to fourfold more active than the other fluoroquinolones. Overall, AM-1155 exhibited more potent in vitro activity against both quinolone-resistant and quinolone-susceptible isolates of N. gonorrhoeae than tosufloxacin and fleroxacin. In ciprofloxacin treatment failures of gonorrhea at single doses of 500 mg. MICs for the causative organisms have ranged from 1.0 to 16.0 micrograms/ml. The MICs of AM-1155 for the isolates harboring quinolone resistance-associated genetic alterations, including strains exhibiting ciprofloxacin MICs of 2.0 and 8.0 micrograms/ml, still ranged from 0.03 to 1.0 microgram/mL A single-dose study in humans has demonstrated higher peak serum concentrations and longer half-lives of AM-1155, resulting in the AUC0 00 values of AM-1155, which are threefold greater than those of ciprofloxacin at the single doses of 400 and 600 mg. Because of its potent in vitro antimicrobial activity and advantageous pharmacokinetic behavior, AM-1155 may be a clinically useful agent for treating gonorrhea including that caused by quinolone-resistant strains. PMID- 9209782 TI - In vitro antimicrobial activity of the thiazolyl peptide antibiotic MDL 62,879 (GE2270 A). AB - Compound MDL 62,879 (GE2270 A) is a thiazolyl peptide antibiotic that appears to inhibit aminoacyl-tRNA binding to elongation factor Tu. In the present study, it was shown that MDL 62,879 broth microdilution MIC values were generally 2-4 doubling dilutions lower in the presence of 0.02% bovine serum albumin. Using US clinical isolates and BSA-supplemented media, MDL 62,879 was more active than teicoplanin and vancomycin against the staphylococci and glycopeptide-resistant and glycopeptide-susceptible enterococci and equally active against the streptococci. Broth microdilutions MIC values were not appreciably affected by inoculum concentrations of 5 x 10(4) to 5 x 10(8) cfu/ml or in the presence of 3.5% human serum albumin. PMID- 9209781 TI - In vitro time-kill curves of cefepime and cefpirome combined with amikacin, gentamicin or ciprofloxacin against Klebsiella pneumoniae producing extended spectrum beta-lactamase. AB - Extended-spectrum beta-lactamases (ESBLs) are found in numerous Enterobacteriaceae, mainly in Klebsiella pneumoniae. We investigated the pharmacodynamics of two new extended-spectrum cephalosporins, cefepime and cefpirome, alone and combined with either amikacin or gentamicin or ciprofloxacin by means of time-kill curves against ESBL-producing, aminoglycoside-resistant K. pneumoniae. When used alone, cefepime (8 and 16 mg/l) resulted in a 2 and 3 log decrease at 6 h, respectively, but at 24 h regrowth occurred. The combination of cefepime (8 mg/l) with amikacin (4 mg/l) resulted in a 4 log decrease at 6 h, but there were no surviving bacteria at 6 h when combined with amikacin (8 mg/l). The combination of cefepime (16 mg/l) with gentamicin (4 mg/l) resulted in a 4 log decrease in 24 h. The antimicrobial combination of cefepime (32 mg/l) with ciprofloxacin (2 mg/l) resulted in a 4 log decrease in 24 h. Cefpirome (8 mg/l) induced a 2 log decrease at 4 h; 32 mg/l cefpirome resulted in a 3 log decrease followed by regrowth at 24 h. The regrowth observed in the late phase with cefpirome alone disappeared when combined with aminoglycoside. When cefpirome (32 mg/l) was used in combination with ciprofloxacin (1 mg/l), it resulted in a 4 log decrease in 24 h. PMID- 9209783 TI - Rifabutin for experimental pneumococcal meningitis. AB - Rifabutin is a lipophilic antibacterial with high in vitro activity against many pathogens involved in bacterial meningitis including pneumococci. Resistance to beta-lactam antibiotics in pneumococci is not associated with a decreased sensitivity to rifabutin (30 strains from Germany with intermediate penicillin resistance; MIC range of penicillin: 0.125-1 mg/l, MIC of rifabutin: < 0.008 0.015 mg/l). Rifabutin at doses of 0.625, 1.25, 2.5, 5 and 10 mg/kg/h i.v. was investigated in a rabbit model of meningitis using a Streptococcus pneumoniae type 3 (MIC/MBC of rifabutin: 0.015/0.06 mg/l). The bacterial density in CSF at the onset of treatment was 7.3 +/- 0.6 log CFU/ml (mean +/- SD). Rifabutin decreased bacterial CSF titers in a dose-dependent manner [delta log CFU/ml/h (slope of the regression line log CFU/ml vs. time) at a dose of 0.625 mg/kg/h: 0.16 +/- 0.06 (n = 3), at 1.25 mg/kg/h: -0.20 +/- 0.12 (n = 4), at 2.5 mg/kg/h: 0.24 +/- 0.04 (n = 4), at 5 mg/kg/h: -0.31 +/- 0.10 (n = 8), and at 10 mg/kg/h: 0.29 +/- 0.10 (n = 5)]. At high doses rifabutin was as active as ceftriaxone at 10 mg/kg/h (delta log CFU/ml/h: -0.29 +/- 0.10, n = 10). Two and 5 h after initiation of therapy, CSF TNF-alpha activities were lower with rifabutin 5 mg/kg/h than with ceftriaxone (medians 2 vs. 141 U/ml, p = 0.005 at 2 h; median 51 vs. 120 U/ml 5 h after initiation of therapy, p = 0.04). This did not result, however, in a decrease of indicators of neuronal damage. In conclusion, intravenous rifabutin was bactericidal in experimental pneumococcal meningitis. Provided that a well-tolerated i.v. formulation will be available it may qualify as a reserve antibiotic for pneumococcal meningitis, in particular when strains with a reduced sensitivity to beta-lactam antibiotics are the causative pathogens. PMID- 9209784 TI - Effects of midazolam on the differentiation of murine myeloid leukemia cells. AB - The effects of midazolam (MID) on the in vitro growth and differentiation of two murine myeloid leukemia WEHI 3B (JCS) and M1 cells were studied. MID inhibits the proliferation of both M1 and JCS cells in a dose-dependent manner. At the concentration of 10 micrograms/ml, MID was found to induce both monocytic and granulocytic differentiation of the JCS but not M1 cells. Induction of morphological differentiation of the JCS cells was also associated with the enhanced expression of the differentiation antigens Mac-1, F4/80, and Gr-1 for the cells. Results from mRNA phenotyping experiments also indicated that the expression of tumor necrosis factor (TNF-alpha) and neutrophil-specific J11d differentiation marker was significantly upregulated in MID-treated JCS cells. In addition, the phagocytic activity of MID-treated JCS cells was increased towards opsonized yeast cells. Results from this investigation suggested that MID may be used as an inducer for further study on the mechanisms of differentiation in these myeloid leukemia cells. PMID- 9209785 TI - Dose response and time course of carboplatin-induced micronucleated polychromatic erythrocytes in the cat: implications for combination carboplatin chemotherapy. AB - The dose response and time course of micronucleated polychromatic erythrocytes (mPCE) in cat peripheral blood induced by various doses (150-250 mg/m2) of carboplatin in vivo was determined. The data indicate that carboplatin produced a significant (p < 0.05) dose-dependent increase in the number of mPCE over baseline values; however, the times following carboplatin administration when mPCE were first observed differed significantly (p < 0.05) between the three carboplatin dose groups. mPCE were present in significantly greater numbers (p < 0.05) on smears at an earlier time interval following a single carboplatin dose of 150 mg/m2 than for a dose of either 200 or 250 mg/m2. The peak number of mPCE occurred on days 7, 14 and 17.5 following administration of a carboplatin dose of 150, 200 and 250 mg/m2, respectively. The pattern of time course delay following carboplatin administration suggests that the block of erythropoietic stem cells in G2 is dose dependent. Indeed, the administration of carboplatin arrested the cell cycle in the G2 phase and, at higher doses, diminished the number of cycling erythroid precursor cells. mPCE were apparent in blood smears only after recovery from this arrest and resumption of replication. This observation has implications for the scheduling of carboplatin administration when used in combination with other anticancer drugs. PMID- 9209786 TI - Significant survival benefit to patients with advanced non-small-cell lung cancer from treatment with clarithromycin. AB - We carried out a randomized study of 49 consecutive patients with unresectable primary lung cancer to determine whether clarithromycin (CAM), a 14-membered ring macrolide, can improve outcome. A total of 49 patients (42 patients with non small-cell lung cancer and 7 patients with small-cell lung cancer) had received prior chemotherapy, radiotherapy or both during their hospital stay. They were randomly allocated into two study groups on the first visit after discharge: 25 patients (22 patients with non-small-cell lung cancer, 3 patients with small-cell lung cancer) were assigned to receive CAM (400 mg/day, orally), and 24 patients (20 patients with non-small-cell lung cancer, 4 patients with small-cell lung cancer) did not receive CAM. CAM treatment after randomization was open and the treatment was to be continued as long as the patients could tolerate CAM. There was no significant difference in the median survival time for small-cell lung cancer between the CAM group and the non-CAM group. However, CAM treatment significantly increased the median survival time for non-small-cell lung cancer patients, the median survival for the CAM group was 535 days and that for the non CAM group was 277 days. Analyses of prognostic factors showed that only treatment with CAM was predictive of longer survival for non-small-cell lung cancer, and other tested covariates had no effects on the prognosis. There were no remarkable side effects observed in the CAM group throughout treatment. We conclude that long-term treatment using CAM is beneficial for unresectable non-small-cell lung cancer patients and that it can increase the median survival of patients with advanced disease. PMID- 9209787 TI - Comparison of ceftibuten versus amoxicillin/clavulanate in the treatment of acute exacerbations of chronic bronchitis. AB - The efficacy and tolerability of once- or twice-daily ceftibuten (400 mg daily) were compared with three-times daily amoxicillin/clavulanate (AMX/CA, 500 mg/125 mg) in the treatment of acute exacerbations of chronic bronchitis (AECB) in an open, parallel-group 10- to 14-day study in 443 patients. Patients were assessed at baseline and on days 5, 10-14 and after 4-6 weeks of treatment, and the clinical response defined as cured, improved, stabilized or failed. Clinical efficacy between the 3 groups was equivalent (p = 0.002) with 90% of patients in each group responding to treatment (cured or improved) and the incidence of complete cures (with no clinical signs of relapse) was also equivalent. In conclusion, this study showed that ceftibuten is clinically equivalent to a standard regimen of amoxicillin/clavulanate in the treatment of AECB, including those patients infected with Streptococcus pneumonia. Ceftibuten was better tolerated than AMX/CA and was associated with significantly fewer gastrointestinal side effects. Furthermore, once-daily was a well tolerated and effective as twice-daily ceftibuten. PMID- 9209789 TI - Assessment of a biuret method without concentration step for total protein determination in cerebrospinal fluid. PMID- 9209788 TI - The acylation stimulating protein pathway: clinical implications. AB - OBJECTIVES: The present review will focus particularly on acylation stimulating protein (ASP) and its role in adipose tissue. Two issues will be addressed (1) in vitro biochemical characterization of ASP in cell culture studies, and (2) in vivo clinical relevance for normal physiology and in pathological conditions. CONCLUSIONS: Fat is In! There can be no question that in recent years fat tissue has become recognized as more than just a passive storage site. It is a metabolically active tissue that, under normal conditions, allows the efficient clearance of triglyceride and glucose for storage as energy. Under abnormal conditions, adipose tissue dysfunction is associated with obesity, diabetes and coronary heart disease. Adipose tissue function may be controlled by many factors. PMID- 9209790 TI - Development of a direct DNA sequencing method for detecting heterozygous mutations of the human lipoprotein lipase gene. AB - OBJECTIVE: The purpose of this study was to develop an improved method of direct DNA sequencing, which makes it possible to identify heterozygous mutations of the lipoprotein lipase (LPL) gene in order to understand the underlying genetic disorder of type IV hyperlipoproteinemia. METHODS AND RESULTS: The direct sequencing method was improved by devising primers for amplifying the LPL gene and for sequencing DNA amplified by the polymerase chain reaction (PCR)T since the reported base sequences of the introns flanking exons of the LPL gene were limited to 40 bases. Improvement was achieved by attaching nine additional bases to both the PCR amplification primer and sequencing primer, and by optimizing the Tm value of the sequencing primers by adjusting the sequence of the nine extra bases. Use of the sequencing primers having suitable Tm values (48 degrees C-58 degrees C) made it possible to reduce nonspecific bands on the sequence ladder pattern and to identify heterozygous mutation sites in LPL gene exons 5 and 6 as model cases. CONCLUSION: Our improved direct sequencing method is useful for identifying heterozygous mutation sites in human LPL gene exons and splicing consensus regions. PMID- 9209791 TI - Elevated sulfatide excretion in compound heterozygotes of metachromatic leukodystrophy and ASA-pseudodeficiency allele. AB - OBJECTIVE: Use of sulfatide excretion in differentiating MLD/PD-heterozygotes from MLD-patients and PD/PD-homozygotes. DESIGN AND METHODS: Sulfatide was extracted from urine sediment with chlorotom/methanol (2:1, v/v). The quantity of sulfatide was measured densitometrically (lambda = 580 nm) after thin-layer chromatography. ASA and beta-galactosidase activities were assayed enzymatically. RESULTS: MLD/PD-heterozygotes excreted sulfatide in the range of 4.8-36.3 nmol/mg lipid (mean +/- SD = 17.8 +/- 10.7), whereas sulfatide in MLD-patients ranged from 74.3-411.6 nmol/mg lipid (mean +/- SD = 184.5 +/- 130.8) and in PD/PD hormozygotes sulfatide excretion remained in normal range of 0.0-5.9 nmol/mg lipid (mean +/- SD = 1.64 +/- 2.12). ASA activities in these groups were very low or lowered. CONCLUSIONS: The quantitative measurement of sulfatide in urine allows differentiation between MLD/PD-heterozygotes and MLD-heterozygotes, as well as between MLD/PD-heterozygotes with very low ASA activity and MLD-patients or PD/PD-hormozygotes. The quantitative measurement of sulfatide in urine differs between MLD-carriers and controls. PMID- 9209792 TI - Screening for Down syndrome during first trimester: a prospective study using free beta-human chorionic gonadotropin and pregnancy-associated plasma protein A. AB - OBJECTIVES: Early screening for Down syndrome is desirable so that more time is left for intervention in the event of a positive test. In retrospective first trimester studies, maternal serum free beta-human chorionic gonadotropin and pregnancy-associated plasma protein A have been reported as useful markers. Our objective was to confirm these results in a prospective study carried on an unselected population. DESIGN AND METHODS: In a cohort of pregnant women recruited prospectively between 9 and 13 weeks' gestation, we measured maternal free beta-human chorionic gonadotropin and pregnancy-associated plasma protein A in all affected pregnancies and 500 representative uneffected pregnancies. Serum concentrations were transformed to multiples of the median value in normal pregnancies of the same length of gestation, and rates of detection of various combinations of the markers were estimated by multivariate analysis. RESULTS: Down syndrome was observed in 18 fetuses from the 10, 160 women recruited. Levels of free beta-human chorionic gonadotropin were elevated in affected pregnancies with an overall median value 1.8 times the median of women with normal pregnancies while pregnancy-associated plasma protein A was significantly diminished (0.51 multiples of the median). At a fixed false-positive risk of 10%, 33% (11-55), 50% (27-73), 44% (11-67), and 67% (45-89) of the affected pregnancies would have been detected (95% CI) with maternal age alone or combined with with free beta-human chorionic gonadotropin, pregnancy-associated plasma protein A or both, respectively. CONCLUSIONS: We confirm in a prospective noninterventional study that maternal serum free beta-human chorionic gonadotropin and pregnancy-associated plasma protein A can be used in the first trimester of pregnancy to screen for Down syndrome with a performance similar to second trimester screening programs. PMID- 9209793 TI - Use of cardiac markers as assessed by outcomes analysis. AB - OBJECTIVES: This article will describe the outcomes studies that have been performed or are needed in relation to biochemical markers in coronary artery diseases (CAD). METHODS AND RESULTS: Studies in five major areas are reviewed: the need for emergency department (ED) chest pain centers and the role of cardiac markers; impact of cardiac marker testing frequency on length of stay (LOS); interpretation of cardiac troponins T and I for risk stratification of cardiac patients with unstable angina (UA); serum markers for determining the success of intravenous thrombolytic therapy following acute myocardial infarction (AMI), and its role in rescue percutaneous transluminal coronary angioplasty (PTCA); and need and criteria for implementation of new cardiac tests. CONCLUSIONS: Chest pain centers reduce unnecessary admissions and costs for AMI rule outs. Laboratories must perform testing on a stat basis for rapid rule out of AMI. Stat testing will also result in a reduction in hospital LOS for patients who rule in for AMI. For UA patients, studies are needed to determine how results of cardiac markers can be used to improve cardiac outcomes. Serial measurements of myoglobin offer the earliest discrimination for successful reperfusion, and should be used if rescue PTCA becomes important therapeutically. New markers for early diagnosis are needed to complement tests such as myoglobin and CK-MB isoforms. Markers that assess early pathophysiologic events of AMI such as inflammation, thrombosis, and pre-necrosis ischemia have the most promise. PMID- 9209794 TI - Correlation between blood antioxidant levels and lipid peroxidation in rheumatoid arthritis. AB - OBJECTIVES: To investigate the relationship between lipid peroxidation and certain antioxidant parameters in the blood of rheumatoid arthritis patients. METHODS AND RESULTS: In the present study, significantly increased lipid peroxidation, measured as malondialdehyde (MDA), was demonstrated in the plasma of rheumatoid arthritis patients (p < 0.01). The activities of erythrocyte antioxidant enzymes, superoxide dismutase and catalase remained unaltered. However, erythrocyte glutathione and plasma ceruloplasmin levels were significantly higher in patients (p < 0.001). Moreover, a positive correlation was also observed between these two parameters and MDA levels in the patient group but not in controls. Interestingly, a significant positive correlation also existed between red cell glutathione and plasma ceruloplasmin levels. CONCLUSION: These results suggest that increased oxidant stress present in rheumatoid arthritis may lead to compensatory changes in the levels of some antioxidants, viz. glutathione and ceruloplasmin. These changes, in turn, may provide additional protection against lipid peroxidation in rheumatoid arthritis. PMID- 9209796 TI - Strategies to promote rational clinical chemistry test utilization. PMID- 9209795 TI - Prostatic cancer: hospital-based prostate specific antigen screening. PMID- 9209797 TI - Autoimmune vasculitis: a good clinical and basic exercise. PMID- 9209798 TI - Cutaneous manifestations in systemic vasculitis. PMID- 9209800 TI - Antiendothelial cell antibodies (AECA) in patients with uveoretinitis. AB - AECAs have been found in 26% of patients with uveoretinitis in studies arising from three different laboratories, and their presence cannot simply be explained by coexisting extraocular disease. There is little correlation with ocular disease activity or other markers of systemic inflammation and vascular damage that can be found in this group of patients, but this lack of correlation has also been found in studies of more widespread inflammatory diseases. The changes found in the peripheral blood of patients with uveoretinitis are the result of a mixture of acute and chronic inflammation, reactions to coexisting tissue damage, as well as predisposing abnormalities of inflammation and hemostasis. Even patients with similar clinical appearances are unlikely to be pathologically homogeneous, and the reasons for the presence of AECA are likely to be various. Some patients may demonstrate a heightened antibody response to endothelium damaged by unknown mechanisms, whereas others may develop cytotoxic AECA as an integral part of the inflammatory process. The majority of serum samples with AECA demonstrated antibody-dependent cell-mediated cytotoxicity, but this potentially pathogenetic mechanism was only demonstrable in a minority of patients. It is unlikely that IgM AECA or complement-mediated cytotoxicity is a relevant mechanism of vascular damage in this group of patients. A subgroup of patients may be genetically predisposed to produce excess autoantibodies in response to tissue damage caused by a wide variety of insults. Sawyerr et al.(36) has suggested that increased serum levels of agalactosyl IgG may account for some of the AECA binding found in chronic inflammatory diseases: we have also found changes in agalactosyl IgG in patients with active isolated uveoretinitis (40), but levels did not correlate with levels of IgG AECA (unpublished results). Further longitudinal studies will be necessary on each subgroup of patients in order to determine the true clinical significance of these findings. PMID- 9209799 TI - ANCA-associated diseases and silica exposure. PMID- 9209801 TI - Antiendothelial cell antibodies (AECA) in systemic lupus erythematosus (SLE). PMID- 9209802 TI - Renal involvement in primary vasculitides. PMID- 9209805 TI - Autoimmune disease in pregnancy. Systemic lupus erythematosus and antiphospholipid syndrome. AB - Autoimmune diseases occur most commonly in women of childbearing age. Over 70% of individuals with autoimmune diseases are women, though some conditions (e.g., ankylosing spondylitis) are more common in men. Many authorities have focused on sex hormones as an explanation for the relatively high proportion of females affected by autoimmune diseases. This article covers systemic lupus erythematosus, the prototypical autoimmune disease, and antiphospholipid syndrome, which is associated particularly with pregnancy loss. PMID- 9209806 TI - Infertility and subsequent pregnancy. AB - Although the incidence of fertility has not increased over the last several decades, the number of patients seeking treatment has increased, especially among older women who have delayed childbearing for various social reasons. During this postponement they are more likely to develop pelvic pathology including endometriosis and uterine myomas and to have been exposed to more encounters with sexually transmitted diseases, all of which contribute to infertility. In addition, it is now well recognized that natural fecundity declines rather dramatically after age 35 years, as a function of the natural depletion of oocytes and decreased ovarian follicular function. Modern endoscopic surgical procedures may be performed to remove myomas, uterine septa, and intrauterine adhesions, but obstetric complications following these operations can occur. Treatment of ovulatory dysfunction can result in multiple pregnancies despite careful monitoring. Patients who fail conventional infertility treatment may now successfully conceive using artificial reproductive techniques, including the use of donor oocytes. This may also result in multifetal gestations and their attendant clinical risks. These aggressive methods of treatment have also led to a marked increase in the occurrence of heterotopic pregnancies, which can be a diagnostic challenge. Infertility patients who conceive should be considered at higher risk for pregnancy complications. A careful history of their fertility treatment and the underlying factors should be reviewed by the obstetric team. Anticipation of known complications will usually result in timely intervention and the successful conclusion of the pregnancy. PMID- 9209807 TI - Pregnancy and intercurrent diseases of the urogenital tract. AB - Pregnancy, labor, delivery, and the puerperium cause many changes in the urinary and genital tracts. The management of lower urinary tract symptoms, disease, and genital prolapse during and after pregnancy is controversial. Patients treated surgically for incontinence, genital prolapse, and lower urinary tract reconstruction present a challenge to the obstetrician and other doctors caring for them during pregnancy. This article reviews the literature on the effect of pregnancy, labor, delivery, and the puerperium on lower urinary and genital tract disease. Preventive methods to reduce subsequent pelvic floor muscle damage and urinary and fecal incontinence are reviewed. The management of pregnant women with antecedent urinary and genital tract abnormalities also are summarized. PMID- 9209808 TI - Preinvasive and invasive breast and cervical cancer prior to or during pregnancy. AB - In conclusion, cervical and breast cancer are the two most frequently encountered malignancies during pregnancy. Although cervical cancer generally is diagnosed in its early stages, breast cancer tends to be discovered in more advanced stages. Therefore, the physician must have a high index of suspicion and must aggressively pursue the diagnosis of breast masses in pregnant women to try to detect the disease as early as possible. Premalignant lesions of the cervix also must be evaluated thoroughly. Therapy for these two cancers is similar to the treatment of nonpregnant women, with some modifications made due to fetal considerations and informed maternal desires. Pregnancy does not affect the progression or prognosis of either disease significantly. Pregnancy following the treatment for cervical cancer is unlikely, if not impossible, because current standard therapies will render these women infertile. There have, however, been some recent reports describing trachelectomy followed by laparoscopic lymph-node dissection for the treatment of early-stage cervical cancer in an attempt to preserve future fertility. Successful pregnancies are possible following the treatment of breast cancer and do not influence recurrence. PMID- 9209803 TI - Pulmonary vasculitis: classification, clinical features, and management. PMID- 9209809 TI - Pregnancy in the chronically hypertensive patient. AB - The majority of patients with mild chronic hypertension have successful pregnancy outcomes. Most perinatal morbidity is secondary to superimposed preeclampsia. Antihypertensive therapy does not appear to significantly affect pregnancy outcome, nor the incidence of superimposed preeclampsia in mild chronic hypertensives. The maternal and fetal risks are considerably higher for severe chronic hypertension and for those patients with target organ disease. These patients ideally should be counseled regarding their risks prior to pregnancy. Antihypertensive therapy should be instituted at diastolic pressures greater than or equal to 100 mm Hg. PMID- 9209810 TI - Asthma and allergy in pregnancy. AB - Rhinitis is extremely common during pregnancy, and asthma is one of the most common potentially serious medical problems to complicate pregnancy. Cutaneous allergy (urticaria/angioedema and eczema) also may occur during pregnancy. All of these entities may worsen with pregnancy in some patients and appear to improve in others. Uncontrolled asthma may directly threaten the fetus, and morbidity from the other illnesses may indirectly affect pregnancy through an effect on eating, sleeping, or emotional well-being. Appropriate diagnosis, avoiding triggering factors when possible; appropriate use of pharmacotherapy; and, when indicated, allergen immunotherapy usually allow these chronic conditions to be controlled during pregnancy so as to optimize both the health of the mother and that of her baby. PMID- 9209804 TI - Mixed cryoglobulinemia as a model of systemic vasculitis. AB - Leukocytoclastic vasculitis is the dominant lesion of mixed cryoglobulinemia (MC). The high prevalence of antibodies to hepatitis C virus (HCV) in association with the higher concentration of HCV RNA genomic sequences in the cryoglobulins suggests a close relationship between MC and HCV infection and strongly supports the view that this virus plays a key role in causing vascular damage. Analysis of the composition of immune complexes (ICs) provides evidence that cryoglobulins include virions mostly bound to IgG that is specifically reactive with HCV related proteins, which in turn are crosslinked by monoclonal IgM with rheumatoid factor (RF) activity, frequently bearing the WA crossidiotype (XId). This structure is similar (if not identical) to that of circulating ICs from HCV infected patients without cryoglobulins, suggesting that the virus may be directly responsible for the production of WA RF. Evidence for the role of circulating cryoproteins in the pathogenesis of cutaneous and renal vasculitis stems from the demonstration of HCV-related proteins and/or HCV RNA genomic sequences in the vessel wall of patients with MC. Our data indicate that endothelial cells are fully susceptible to infection by and replication of HCV, and support the contention that they serve as sufficient targets for the binding of HCV proteins expressed on the cell surface to serum immunoglobulins. The in situ demonstration of IgM RF WA XId adds further evidence that RF of the WA group participates in the development of vasculitis and probably stabilizes the binding of IgG antibodies. Lymphocytes may be crucial in the infection of endothelial cells by acting as a circulating viral reservoir. After encouraging initial results, controlled trials have defined the substantive efficacy of IFN-alpha in the treatment of MC. A response of IFN can be achieved in more than 50% of patients and includes improvement of cutaneous vasculitis and renal function. This clinical response is accompanied by a reduction in hepatitis C viremia, serum cryoglobulin concentration, and IgM RF synthesis. However, almost 80% of responders eventually have a clinical and biochemical relapse. Additional studies are required to improve the outcome and extension of this therapy, define the best candidates, and indicate the situations in which it is needed. PMID- 9209811 TI - Maternal nutrition and the outcome of pregnancy. A renaissance in research. AB - Research over the past decade indicates that aspects of maternal nutrition may play a greater role in immediate and long-term health of offspring than was thought previously. The effects of nutritional exposures on outcomes likely depend primarily on the timing of insults during pregnancy and may not have to be severe to have a long-term impact on health. If the rate of change in knowledge of the effects of maternal nutritional exposures on outcomes continues at its current pace, work initiated in the past decade may well be looked back upon as representing the renaissance of maternal nutrition research. PMID- 9209813 TI - Congenital cardiac disease and pregnancy. AB - Advances in the detection and treatment of congenital heart disease has made it possible for children born with congenital heart disease to reach adulthood. Pregnancy often is feasible in the woman born with congenital heart disease; however, the type of lesion, surgical repair, and residual hemodynamic state will determine the success of the pregnancy. Knowledge of the specific congenital lesion and type of surgical repair helps the physicians caring for these women to assess them prior to pregnancy to be able to anticipate and treat complications that may occur in the settling of their chronic disease. PMID- 9209812 TI - Maternal genetic disease and pregnancy. AB - This article reviews classic genetic diseases and illustrates the effect of an existing maternal condition on maternal and fetal well-being during pregnancy. The management issues discussed may serve as a framework for treatment when reproductive issues are encountered with other maternal genetic diseases. PMID- 9209814 TI - Pregnancy complicated by chronic renal disease. AB - Regardless of the cause or degree of renal insufficiency, some common themes have run through the conclusions of many investigators studying chronic renal disease and pregnancy: fetal survival, once thought rare, now appears to be commonplace; maternal renal function, previously thought unlikely to survive a pregnancy, is not adversely affected in the majority of well-managed pregnancies, though the risk of deterioration is still considerable; of all the parameters that can be followed and influenced, maintenance of normotension is the most important. The improvements in maternal and fetal outcome clearly are due to improvements in postnatal care for the neonate, as well as an increased understanding of important risk factors leading to better antepartum maternal management. These women should all receive preconceptual counseling and optimization of their disease condition. Those choosing to become or continue pregnancy should receive their prenatal care from a team including perinatologists, nephrologists, and neonatologists. Only through a tightly coordinated, team approach can the meticulous care offering the opportunity for successful fetal outcome and minimization of maternal risk be provided. PMID- 9209816 TI - The athletic trainer's perspective. AB - This article provides a comprehensive understanding of the qualifications and responsibilities of an athletic trainer. It describes the athletic trainer's role in conditioning, injury prevention, and treatment of athletic injuries. The advantages of different types of flexibility, running, and strength training are also included. In addition, the importance of diet and fluid replacement are explained with the guidelines to follow, and treatment and rehabilitation programs for the injured athlete are described. PMID- 9209815 TI - The great balancing acts. The pregnant woman, placenta, fetus, and infectious agents. AB - This article conceptualizes the various balancing acts between the pregnant woman, the placenta, the fetus, and the infectious agents. Despite the very large number of infectious insults during pregnancy, the outcome of most interactions usually is a normal newborn. We have identified only one general immune defect of the fetus and neonate: The inability to respond to polysaccharide antigens; yet, a similar defect also is found with certain polysaccharides in older children and adults. The capacity of the neonate to control severe life-threatening diseases with most infectious agents and the ability of the fetus, when infected in utero, to mount sophisticated, immune responses make it conceptually advantageous to consider the older fetus and the newborn infant as "immunodelayed" rather than as immunodeficient or immature. PMID- 9209819 TI - Soft-tissue injuries and muscle tears. AB - Soft-tissue injuries and muscle tears occur frequently in athletes. The mainstay of treatment in most cases is nonoperative management and aggressive rehabilitation. Most injuries result from direct trauma or contusion or indirect stretch injury. It is important to keep in mind the possibility of other potentially more serious conditions, such as compartment syndrome. More research is needed to define optimal treatment patterns and potential strategies for injury prevention. PMID- 9209818 TI - Contemporary imaging of athletic injuries. AB - The team physician is faced with an array of imaging studies to evaluate the injured athlete, including CT scans, conventional and cross-sectional arthrography, bone scanning, sonography, and MR imaging. An understanding of the clinical usefulness of these examinations requires some knowledge of the physical principles on which they are based. This article presents the fundamental physical principles and summarizes the indications for these tests to the evaluation of common athletic injuries. PMID- 9209817 TI - Ergogenic drugs in sports. AB - This article provides the practicing physician with an account of the commonly used ergogenic substances. Specific agents discussed include the following: stimulants, narcotic analgesics, anabolic-androgenic steroids, beta-blockers, diuretics, growth hormone, other peptide hormones, blood doping, and erythropoietin. PMID- 9209820 TI - Primary care of foot and ankle injuries in the athlete. AB - A thorough knowledge of foot and ankle anatomy is required to allow an accurate and focused examination of the injured athlete. This short review has attempted to educate the treating physician on our approach to foot and ankle injuries commonly seen in athlete. We have tried to elucidate less common injuries that present in similar manner to the more common foot and ankle sprains and strains. PMID- 9209821 TI - Leg injuries. AB - Injuries in the leg span a broad spectrum of patient age and athletic level. Overuse injuries, such as medial tibial stress syndromes and stress fractures, tend to occur in the young athlete, whereas tennis leg usually occurs in the older population. With a few exceptions, most of these injuries can be successfully treated nonoperatively. Particularly with the young athlete, it is important to stress the necessity to rest and avoid activities that would compound the injury. PMID- 9209822 TI - Common athletic knee injuries. AB - Knee injuries continue to be an increasingly common and highly visible problem presenting to the sports medicine physician. The physicians who handle knee injuries will be challenged by patients, coaches, trainers, business agents, the press, and family members with an ever-increasing sophistication of medical knowledge. An understanding of the underlying structure and function of the commonly injured ligaments, menisci, and the patellofemoral joint is discussed. Diagnosis by physical examination is encouraged. Conservative and surgical treatment options are reviewed as are the considerations involved in deciding the time to return to sports. PMID- 9209823 TI - Management guidelines for participation in collision activities with congenital, developmental, or post-injury lesions involving the cervical spine. AB - We believe that the aforementioned management guidelines for participation in collision activities for individuals with congenital, developmental, or postinjury lesions involving the cervical spine have been formulated on the basis of the best information available to date. It is recognized that modifications may occur as more data are collected. We emphasize that these proposed guidelines should be used in the decision-making process in conjunction with such other factors as age, experience, ability of the individual, level of participation, and position played. A most important consideration is the attitude and desire of the individual and his parents following an informed discussion of the problem with particular regard to potential risks. PMID- 9209824 TI - Athletic head injuries. AB - Recent studies have shown a decrease in mortality from head and neck injuries, especially in American football. This has resulted from rule changes and their enforcement, equipment modification, improved coaching and training techniques, and educational programs for neck injuries. Morbidity data, however, are not as complete, particularly as they apply to concussion, the most frequent type of head injury in contact sports. Questions on this condition that still need to be answered before a sound medical disposition can be made are the possible cumulative damage from repeated concussions, and whether one concussion renders a player more susceptible to a second. Currently, decisions on when to allow a football player to return to a game or participate in future contests are arbitrary and based primarily on the experience of the team physician. Certainly, further studies are essential before these decisions can be based on sound scientific data. Thus, the pioneer work of Richard Schneider needs to be continued. PMID- 9209825 TI - Sports-related facial injuries. AB - Facial injuries suffered by the athlete will range from very minor to extremely complex. Regardless, all injuries can be accurately diagnosed if one is familiar with normal anatomy and conducts a thorough physical examination. Radiographs are then obtained as directed by the physical examination. Specific and timely treatment directed at correcting functional and cosmetic deformities will help to avoid lifelong disability and deformity. With adherence to these guidelines, many of the injuries suffered by athletes can be safely treated by a variety of physicians. More complex soft-tissue and bone injuries should be referred to a surgeon familiar and comfortable with facial plastic and reconstructive surgery. PMID- 9209826 TI - The neuropsychology of spatial cognition in the rat. AB - This article provides a review of the neural mechanisms of spatial cognition in the rat. A survey of the literature shows that the rat has spatial capabilities that can be explained only if one assumes that it possesses a representation of some features of the environment. The scope of such a representation may, however, be more limited than what is implied by the hypothesis of a bird's-eye view of the environment. The best documented spatial ability of the rat is illustrated by its efficiency in performing the water maze navigation task. A review of recent neurobiological data collected while a rat was performing this task suggests that several brain structures make unique contributions to spatial navigation. In particular, the hippocampal formation and the associative (posterior) parietal cortex seem to handle different aspects of navigation and to be differentially involved in the various stages of spatial memory formation. Electrophysiological data support the hypothesis that the hippocampal formation is concerned with rapidly building associative memories of spatial relationships within the environment. In contrast, the associative parietal cortex might be involved in more abstract spatial processing, resulting in a metric representation of spatial information collected during movements. PMID- 9209827 TI - Dopamine D2 receptors in signal transduction and behavior. AB - The dopamine D2 receptor belongs to the family of seven transmembrane domain G protein-coupled receptors and is highly expressed in the central nervous system and the pituitary gland. The binding of dopamine to the D2 receptor is crucial for the regulation of diverse physiological functions, such as the control of locomotor activity and the synthesis of peptide hormones. Two alternatively spliced transcripts are generated from the D2 receptor gene and code for the D2L and D2S isoforms, which are 444 and 415 amino acids in length, respectively. These isoforms exhibit similar pharmacological characteristics and are expressed in the same cell types, with a ratio that normally favors expression of the longer isoform. The D2L isoform differs from D2S by the insertion of 29 amino acids in the putative third intracellular loop of the receptor. This loop is involved in the coupling of the receptor to different G proteins. Experiments have shown that the D2 isoforms have different G-protein-coupling affinities, suggesting that these receptors might serve different functions in vivo. Additionally, this difference in coupling affinity could be a mechanism to amplify the signal transduced by the binding of dopamine to D2 receptors. Important insights into D2 receptor function in vivo have been obtained by knocking out the D2 gene in mice. The Parkinsonian-like phenotype of D2-null mice demonstrates the importance of the D2 receptor for locomotor function. PMID- 9209828 TI - Molecular aspects of cannabinoid receptors. AB - Two cannabinoid receptors are reviewed with regard to their primary structure, ligand-binding properties, and signal transduction systems. Both receptors have been cloned; therefore, the expression of their genes and the functional domains within the proteins can be examined. Binding of tritiated agonists has localized these receptors to the central nervous and immune systems. The CBI receptor is predominantly expressed in brain tissues and is found in both glial elements and neurons; subcellular localization to axons and terminals is evident. This receptor is found in motor, limbic, associative, cognitive, sensory, and autonomic brain structures. CBI receptors modulate the activities of calcium and potassium channels. The CB2 receptor is predominantly expressed in the immune system and is found in spleen, tonsils, thymus, mast cells, and blood cells. Although receptors appear to be involved in cannabimimetic-induced modulation of immune cell function, the receptor subtype that is principally involved in specific effects is difficult to determine because both receptors are often coexpressed in the same cells. Cannabimimetic-induced effects on mast cells and B cells appear, however, to be mediated by CB2 receptors. PMID- 9209830 TI - Use of stem cell factor to mobilize hematopoietic progenitors. AB - Stem cell factor (SCF) is a multipotent growth factor that plays a role in the growth and development of hematopoietic cells, spermatocytes, melanocytes, and mast cells. SCF alone has little direct stimulatory activity on hematopoietic progenitors but acts synergistically with other colony-stimulating factors to potentiate their effects on colony growth. SCF also potentiates the mobilization of hematopoietic progenitor cells into the peripheral blood in response to granulocyte colony-stimulating factor (G-CSF), with or without chemotherapy. The combination of SCF plus G-CSF appears to be a particularly effective mobilization regimen for patients who have been heavily pretreated. Whether engraftment of peripheral blood progenitor cells mobilized by SCF plus G-CSF is superior to that of cells mobilized by G-CSF alone remains unclear. SCF appears to be a necessary requirement for ex vivo expansion of hematopoietic progenitors in both liquid and bioreactor culture systems and will be an important component of future cellular therapies, such as expansion of placental cord blood progenitors and retroviral mediated gene transfer into hematopoietic target cells. PMID- 9209829 TI - Amylin, calcitonin gene-related peptide, calcitonin, and adrenomedullin: a peptide superfamily. AB - The calcitonin gene peptide superfamily consists of calcitonin (CT), calcitonin gene-related peptide (CGRP), and amylin. CT and CGRP derive from the CT/CGRP gene, which is encoded on chromosome 11. Alternative splicing of the primary RNA transcript leads to the translation of CGRP and CT peptides in a tissue-specific manner. CGRP (a 37-amino-acid neuropeptide) and its receptors are widely distributed in the body, and it is the most potent endogenous vasodilatory peptide discovered so far. CT (a 32-amino-acid peptide) is, however, a hormone primarily involved in protecting the skeleton during periods of "calcium stress" such as growth, pregnancy, and lactation. CT derives from the C cells of the thyroid gland and is the most potent peptide inhibitor of osteoclast-mediated bone resorption. Therefore, treatment with CT is highly effective for conditions associated with increased bone turnover such as Paget's disease, osteoporosis, Sudeck's atrophy, and hypercalcemia. Amylin (a 37-amino-acid peptide) is generated from a gene located on chromosome 12 (thought to be an evolutionary duplication of chromosome 11) and shares 46% amino acid sequence homology with CGRP and 20% with human CT. Amylin is predominantly located in the beta cells of the islets of the pancreas and may be involved in the pathogenesis of type II diabetes by deposition as amyloid within the pancreas, leading to beta cell destruction. Adrenomedullin, a recently discovered 52-amino-acid vasoactive peptide from adrenal tissue, shares 24% homology with CGRP and is also a member of this superfamily of peptides. A portion of the B-chain of insulin is strongly homologous to these four peptides. Not only does adrenomedullin (13-52) show 24% amino acid homology with CGRP, it also has a biological activity profile similar to that of CGRP.CGRP, CT, and amylin are related to the insulin gene superfamily of peptides, which may all have diverged from a common ancestral gene during evolution. When the crystallographic- and nuclear magnetic resonance-based molecular modeling of the three-dimensional structure of CGRP, CT, amylin, and adrenomedullin peptides and their receptors is available, it will lead to a greater understanding of the involvement of this family of peptides in pathophysiology. Together, CGRP, CT, amylin and adrenomedullin have overlapping biological effects owing to their structures and cross-reactivity between receptors. I propose that CT, CGRP, adrenomedullin, and amylin belong to a family of G-protein-coupled receptors (an "insulin superfamily" of peptides) and therefore share some of the characteristics of insulin, such as growth factor like effects, and possible interaction at insulin receptor sites as an antagonist. PMID- 9209831 TI - In vivo effects of Mpl ligand administration and emerging clinical applications for the Mpl ligands. AB - The Mpl ligands are a family of closely related hematopoietic growth factors that bind to the thrombopoietin receptor, c-Mpl. In addition to the endogenous Mpl ligand, thrombopoietin, two recombinant Mpl ligands, recombinant thrombopoietin and pegylated megakaryocyte growth and development factor (PEG-MGDF) are under investigation. Endogenous thrombopoietin regulates most of the normal production of platelets but also is essential for the normal development of other lineages. When recombinant thrombopoietin or PEG-MGDF is administered to normal animals or humans, there is a dose-dependent increase in the platelet count but no effect on leukocytes or erythrocytes. When administered following chemotherapy in animal models or humans, Mpl ligands reduce the duration and sometimes the degree of thrombocytopenia. The Mpl ligands also may be effective in reducing the thrombocytopenia of patients with HIV infection, liver disease, myelodysplasia, or after plateletpheresis. PMID- 9209833 TI - Phenotype and engraftment potential of cytokine-mobilized peripheral blood mononuclear cells. AB - Cytokine-mobilized peripheral blood stem cell products are increasingly used for hematopoietic reconstitution after myeloablative therapy. Favorable engraftment kinetics, the ease of harvest, and the large number of CD34+ cells obtained that allow for graft manipulations (ie, tumor cell or T-cell depletion) have made this stem cell source an attractive alternative to marrow. More recent data suggest that in addition to the increased number of CD34 cells, there may be also qualitative differences between leukapheresis products and marrow. In the allogeneic transplantation setting, the one log more T cells contained in granulocyte colony-stimulating factor-mobilized peripheral blood mononuclear cells compared with marrow has not translated into more severe graft-versus-host disease, indicating possible differences in T-cell or accessory-cell function. Whether such differences will compromise graft-versus-leukemia effects and disease-free survival remains to be seen. Nevertheless, it is reasonable to speculate that cytokine-mobilized peripheral blood products may eventually replace marrow as a source for hematopoietic stem cells. However, each new mobilization strategy needs to be evaluated carefully, as comparable increases in CD34 cell numbers may not necessarily affect the same, as yet underlined, qualitative changes that make this product so attractive. PMID- 9209832 TI - Advances in understanding postreceptor signaling in response to granulocyte colony-stimulating factor. AB - Granulocyte colony-stimulating factor (G-CSF) exerts its biologic effects through binding to its receptor expressed on myeloid cells. Like other cytokines, G-CSF induces intracellular protein tyrosine phosphorylation and activates various signaling cascades. Activation of JAK tyrosine kinases and signal transducers and activators of transcription (STAT) proteins as well as activation of the ras-MAP kinase route results in induction of gene transcription. Distinct regions or defined tyrosine residues of the G-CSF receptor cytoplasmic domain are required for complex formation with specific signaling molecules and ultimately regulate proliferation and maturation of myeloid cells. In vivo, administration of G-CSF results in increased numbers of neutrophils in normal individuals, in patients with chemotherapy-induced neutropenia, and in patients with chronic neutropenia. A subgroup of patients with severe congenital neutropenia displayed point mutations in the cytoplasmic region of the G-CSF receptor: These G-CSF receptor mutations might be involved in leukemogenesis in congenital neutropenia. PMID- 9209834 TI - Effects of granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor on the bactericidal functions of neutrophils. AB - The hematopoietic growth factors granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) not only regulate the numbers of circulating neutrophils but also modulate the function of mature cells. Additionally, newly developed neutrophils subsequently released from the bone marrow in response to colony-stimulating factors (CSFs) also have enhanced function. A variety of different functions are affected, including changes in adherence, movement, phagocytosis, priming and stimulation of the respiratory burst, secretion, and degranulation. These effects also can cause increased microbicidal capacity in vitro, ex vivo, and in vivo. Both G-CSF and GM-CSF have such effects on neutrophil function, but there are differences that may result in precise modulation of the immune responses and may have implications for choice of agents for immune-based therapy for different conditions. PMID- 9209835 TI - Use of hematopoietic growth factors in the treatment of acute myelogenous leukemia. AB - Several randomized trials evaluating the effect of hematopoietic growth factors, especially granulocyte colony-stimulating factor (G-CSF) and granulocyte macrophage colony-stimulating factor (GM-CSF), in the treatment of patients with acute myeloid leukemia, recently have been completed. The results of these trials generally show a reduction in the duration of neutropenia with variable results as far as severe infections, antibiotic use, and duration of hospitalization are concerned. Combining the data from the studies and especially from those with higher patient numbers, complete remission rates, event-free survival, and overall survival do not appear to be affected by the use of either G-CSF or GM CSF after induction and consolidation therapy. The use of G-CSF or GM-CSF either before or during induction chemotherapy in an attempt to increase the leukemic cell kill has not resulted in improved response rates or survival. Only a few case reports indicate that G-CSF and GM-CSF might induce terminal differentiation of leukemic blast cells. Recently started trials evaluating the effect of megakaryocyte growth and development factor on platelet recovery indicate that it might not be easy to show a clinical benefit, possibly because of the rather low threshold counts needed for the prevention of overt bleeding. It still remains unclear whether the use of hematopoietic growth factors in these patients is cost effective. PMID- 9209836 TI - Use of colony-stimulating factors in the treatment of neutropenia associated with collagen vascular disease. AB - Chronic neutropenia associated with collagen vascular disease is seen principally with Felty's syndrome complicating rheumatoid arthritis. Multiple recent reports document the efficacy of both granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) in reversing the neutropenia and decreasing the risk of infections in Felty's syndrome. Long-term use of G-CSF appears well tolerated and effective in Felty's syndrome. Of concern, however, have been flares of arthritis and development of leukocytoclastic vasculitis in several patients following the use of colony stimulating factors (CSFs) in Felty's syndrome. The incidence of these complications of CSF therapy appears to be greater in Felty's syndrome than in other disorders. Future studies will need to address the incidence of these side effects, evaluate strategies to reduce risks, and clarify the optimum use of CFSs in Felty's syndrome. PMID- 9209837 TI - Role of granulocyte colony-stimulating factor and granulocyte-macrophage colony stimulating factor in the treatment of patients with HIV infection. AB - The recombinant human colony-stimulating factors, granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are hematopoietic cytokines that increase neutrophil number and enhance their function. In patients with HIV infection, G-CSF and GM-CSF have reversed or prevented neutropenia even during periods of full-dose myelotoxic therapy. Both colony-stimulating factors (CSFs) also have improved defects in neutrophil function in the setting of HIV infection. In non-neutropenic animal models of opportunistic bacterial or fungal infections, use of CSFs has increased survival. Future clinical applications of CSFs may include the adjunctive treatment of specific HIV-related opportunistic infections in addition to an expanding role in the treatment of HIV-associated neutropenia and defects in neutrophil function. PMID- 9209838 TI - Use of granulocyte colony-stimulating factor in the treatment of acute infectious diseases. AB - Infectious diseases continue to be a major cause of morbidity and mortality in the United States. Novel strategies directed at amplifying critical components of the host's immune system may be one potential adjuvant therapy. To date, studies with granulocyte colony-stimulating factor (G-CSF) treatment have demonstrated favorable results, decreasing both morbidity and mortality in a variety of infectious disease models in non-neutropenic hosts. The recent completion of two clinical trials has also provided supportive data that G-CSF is effective in patients with serious infections. Most important, G-CSF does not produce any adverse consequences, which were of concern to some because of the capacity of G CSF to enhance neutrophil production and function. This review focuses on current data suggesting that augmentation of the immune system with G-CSF is a promising new approach to adjuvant therapy in infectious diseases. PMID- 9209839 TI - Safety of concomitant use of granulocyte colony-stimulating factor or granulocyte macrophage colony-stimulating factor with cytotoxic chemotherapy agents. AB - Most studies that use recombinant granulocytopoietic cytokines, such as granulocyte colony-stimulating factor (G-CSF) and granulocyte-macrophage colony stimulating factor (GM-CSF), with the intent of attenuating neutropenia generally have delayed the administration of the cytokine until 24 to 72 hours following completion of chemotherapy. This practice was initiated out of theoretic concern that colony-stimulating factor administration may cycle and differentiate a population of normal cells, thus increasing their susceptibility to cycle specific antineoplastic agents. The theory, in fact, has been substantiated by evidence from several clinical trials of concurrent administration. Thus, simultaneous administration of chemotherapy and G-CSF or GM-CSF should be limited to investigational protocols with scientific objectives, such as cycle compression or malignant cell sensitization. PMID- 9209840 TI - Safety of long-term administration of granulocyte colony-stimulating factor for severe chronic neutropenia. AB - When a new product with huge clinical potential explodes on the scene, the hope is that the benefits far outweigh the risks in long-term administration. After 10 years of clinical use, granulocyte colony-stimulating factor (G-CSF) has lived up to that promise so far. In the context of severe chronic neutropenia, more than 90% of patients have reaped big benefits in terms of improved quality of life and less infection, inflammation, antibiotic use, and hospitalization as well as oropharyngeal ulcers. With long-term use, toxic and adverse events have been catalogued but in general are not clinically troublesome and, aside from occasional adjustment of scheduling and dosing, seldom necessitate stopping therapy. Currently, the topic of intense focus is the phenomenon of malignant myeloid transformation in patients with congenital neutropenia who are receiving G-CSF. Data from the Severe Chronic Neutropenia International Registry have identified 23 of 249 patients with congenital neutropenia who have developed myelodysplasia or acute myelogenous leukemia (MDS/AML) giving a crude rate of about 9% with an average follow-up of 4.5 years. No cases of MDS/AML have occurred in 257 patients with cyclic or idiopathic neutropenia. A critical analysis of the incidence of transformation year by year showed a fairly uniform hazard rate of less than 2% per year, and the risk of MDS/AML after 5 to 6 years of therapy did not appear to be greater than during the first year of therapy. The transformation risk in the congenital cohort must also be viewed in the context of published reports of spontaneous myelodysplasia or acute myelogenous leukemia occurring in these patients in the pre-G-CSF era. Thus, the role of G CSF in malignant conversion is still not clear and requires long-term vigilance and research. G-CSF is still deemed specific therapy for severe chronic neutropenia with a high margin of safety and should be the initial treatment for this family of disorders. PMID- 9209841 TI - Hematopoietic growth factors. PMID- 9209842 TI - Musings on the edge of epidemiology. PMID- 9209843 TI - Collaboration produces a whole that is greater than the sum of its parts. PMID- 9209844 TI - Replication. PMID- 9209845 TI - Features of tree-structured survival analysis. PMID- 9209846 TI - Maternal residential proximity to hazardous waste sites and risk for selected congenital malformations. AB - Using data from two population-based case-control studies, we investigated whether maternal residential proximity to hazardous waste sites increased the risk for neural tube defects, conotruncal heart defects, and oral cleft defects in California. We obtained a residential history by interview for mothers of 507 neural tube defect cases (82.7% of eligible) and their 517 controls (84.6%); and 201 heart cases (84.4%), 439 cleft cases (82.2%), and their 455 controls (72.1%). We identified the locations of 764 inactive hazardous waste sites and systematically collected information on site-related contamination for the subset of 105 National Priority List sites. After controlling for several potential confounders, we found little or no increased risk for maternal residence in a census tract containing a site [odds ratio (OR) = 0.9, 95% confidence interval (CI) = 0.7-1.3 for neural tube defects; OR = 1.3, 95% CI = 0.8-2.1 for heart cases; OR = 1.2, 95% CI = 0.8-1.8 for clefts], but elevated risks for neural tube defects (OR = 2.1, 95% CI = 0.6-7.6) and heart defects (OR = 4.2, 95% CI = 0.7 26.5) for maternal residence within 1/4 mile of a National Priority List site. Furthermore, we observed elevated ORs (> or = 2.0) for neural tube defects and heart defects in association with maternal residence within 1 mile of National Priority List sites containing selected chemical contaminants. Among controls, only 0.6% and 4.4% lived within 1/4 mile and 1 mile of a National Priority List site, respectively, resulting in imprecision in risk estimation. PMID- 9209847 TI - Congenital malformation and maternal occupational exposure to glycol ethers. Occupational Exposure and Congenital Malformations Working Group. AB - Glycol ethers are found in a wide range of domestic and industrial products, many of which are used in women's work environments. Motivated by concern about their potential reproductive toxicity, we have evaluated the risk of congenital malformations related to glycol ether exposure during pregnancy as part of a multicenter case-control study, conducted in six regions in Europe. The study comprised 984 cases of major congenital malformations and 1,134 controls matched for place and date of birth. Interviews of the mothers provided information about occupation during pregnancy, sociodemographic variables, and other potential risk factors (medical history, tobacco, alcohol, drugs). A chemist specializing in glycol ethers evaluated exposure during pregnancy, using the job description given by the mother, without knowledge of case or control status. We classified malformations into 22 subgroups. The overall odds ratio (OR) of congenital malformation associated with glycol ether exposure was 1.44 [95% confidence interval (CI) = 1.10-1.90], after adjustment for several potential confounders. The association with exposure to glycol ethers appeared particularly strong in three subgroups: neural tube defects (OR = 1.94; 95% CI = 1.16-3.24), multiple anomalies (OR = 2.00; 95% CI = 1.24-3.23), and cleft lip (OR = 2.03; 95% CI = 1.11-3.73). In this last subgroup, risk, especially of an isolated defect, tended to increase with level of exposure. PMID- 9209848 TI - Air pollution and hospital admissions for respiratory causes in Minneapolis-St. Paul and Birmingham. AB - We investigated the association between air pollution and hospital admissions for chronic obstructive pulmonary disease and pneumonia among the elderly in Minneapolis-St. Paul, MN, and Birmingham, AL, over the period January 1, 1986, to December 31, 1991. Pollutants included in our analyses were PM10 (particulate matter less than 10 microns in aerodynamic diameter), SO2, NO2, O3, and CO in Minneapolis-St. Paul, and PM10, O3, and CO in Birmingham. After adjusting for temperature, day of week, season, and temporal trends, we found little evidence of association between air pollution and hospital admissions for respiratory causes in Birmingham. In contrast, we found that air pollution was associated with hospital admissions for respiratory causes in Minneapolis-St. Paul. Among the individual pollutants, O3 was most strongly associated with admissions (estimated increase in hospital admissions associated with a 15-parts-per-billion increase in O3 on the previous day = 5.15%; 95% confidence interval = 2.36 7.94%), and this association was robust in the sense that it was little affected by the simultaneous consideration of other pollutants. PM10, SO2, and NO2 were also associated with hospital admissions, although none could be singled out as being more important than the others. PMID- 9209849 TI - Air pollution and hospital admissions for cardiovascular disease in Tucson. AB - Several recent studies have reported associations between short-term changes in both inhalable particles (PM10) and carbon monoxide and cardiovascular hospital admissions. Here, I seek to replicate those findings in a location where sulfur dioxide concentrations are low and poorly correlated with PM10, and where PM10 concentrations peak in the winter when ozone is lowest. This setting allows the opportunity to separate the effects of different air pollutants. I constructed daily counts of admissions to all hospitals in Tucson, AZ, for cardiovascular disease (International Classification of Diseases, 9th revision, codes 390-429) for persons age 65 years and older. I analyzed these admission counts in a Poisson regression, on temperature, humidity, day of the week indicators, and air pollution. I removed long wavelength patterns using a nonparametric smooth function of day of study. I used regression splines to model possible nonlinearities in the dependence of hospital admissions on weather. I then examined sensitivity analyses to control for weather. Both PM10 and carbon monoxide were associated with increased risk of cardiovascular hospital admissions. Admissions increased by 2.75% [95% confidence limits (CL) = 0.52%, 5.04%] for an interquartile range increase (23 micrograms per m3) in PM10 and by 2.79% (95% CL = 0.51%, 5.41%) for an interquartile range increase (1.66 parts per million) in carbon monoxide. These associations were independent and additive. In contrast, I saw little association with sulfur dioxide [increase of 0.14% (95% CL = -1.3%, 1.6%) for an interquartile range increase in exposure], ozone [increase of 0.54% (95% CL = -2.3%, 3.45%)], or nitrogen dioxide [increase of 0.69% (95% CL = -2.3%, 3.8%)]. The air pollution associations were insensitive to control for a potential interaction between temperature and humidity and to control for temperature and humidity on more than 1 day. PMID- 9209850 TI - Obesity and 33-year follow-up for coronary heart disease and cancer mortality. AB - We used tree-structured survival analysis (TSSA), a computer-intensive method of classification, to determine prospectively the relation of the body mass index and the waist-to-calf circumference ratio to coronary heart disease and cancer mortality in 3,155 middle-aged men initially free of these diseases. Applied to coronary heart disease mortality, TSSA identified seven subgroups that differed in profile of risk factors and associated survival. Among the seven subgroups, a small subgroup of older, obese, normotensive men (N = 71) experienced an exceptionally high risk of coronary heart disease deaths over the 33 years of follow-up (34%), similar to the risk of 36% experienced by a larger subgroup (N = 387) of men of similar ages who were less obese and had higher blood pressure levels. We also observed a higher overall risk of coronary heart disease deaths during follow-up (10.3% vs 5.3%) in younger centrally obese men who had low blood pressure levels compared with their counterparts of similar age who were less obese. When applied to mortality from cancer of all sites, TSSA identified five subgroups that differed in survival distributions and profile of risk factors. A subgroup of younger, centrally obese, and ever-smoker men experienced a higher risk of cancer deaths than their counterparts who were less obese (14% vs 8%). Results from these analyses demonstrate the usefulness of a tree-structured analysis for classification of subjects into high- and low-risk survival subgroups. PMID- 9209851 TI - Occupational and residential magnetic field exposure and leukemia and central nervous system tumors. AB - Studies of magnetic field exposure and cancer have focused on either residential or occupational exposure. We conducted a case-control study taking into account both exposure sources. We identified leukemia and central nervous system tumor cases and controls from a population living within 300 m of transmission lines in Sweden. We have previously reported results considering residential exposure alone. Here, we evaluate the effect of occupational exposure and of the combined exposures. We estimated residential exposure through calculations of the magnetic fields generated by power lines. We obtained information about occupation from censuses and linked the occupations to a job-exposure matrix based on magnetic field measurements. For occupational exposure of > or = 0.2 microT, we estimated the relative risk for leukemia to be 1.7 [95% confidence interval (CI) = 1.1 2.7]. The increased risk was confined to acute myeloid and chronic lymphocytic leukemia. For residential exposure of > or = 0.2 microT, the relative risk for leukemia was estimated at 1.3 (95% CI = 0.8-2.2), with higher risk estimates for acute and chronic myeloid leukemia. We estimated the relative risk for leukemia among subjects highly exposed both at home and at work to be 3.7 (95% CI = 1.5 9.4). These results provide support for an association between magnetic field exposure and leukemia. Relative risks for nervous system tumors were close to unity. PMID- 9209852 TI - A method for timely assessment of influenza-associated mortality in the United States. AB - Influenza-associated mortality has traditionally been estimated as the excess mortality above a baseline of deaths during influenza epidemic periods. Excess mortality estimates are not timely, because national vital statistics data become available after a period of 2-3 years. To develop a method for timely reporting, we used the 121 Cities Surveillance System (121 Cities), maintained at the Centers for Disease Control and Prevention, as an alternative data source. We fit a cyclical regression model to time series of weekly 121 Cities pneumonia and influenza deaths for 1972-1996 to estimate the excess pneumonia and influenza mortality and to compare these figures with national vital statistics estimates for 20 influenza seasons during 1972-1992. Seasonal excess mortality based on 121 Cities correlated well with the national data: for 18 (90%) of 20 seasons, our influenza epidemic severity index category approximated the result based on national vital statistics. We generated preliminary severity categories for the four recent seasons during 1992-1996. We conclude that the 121 Cities Surveillance System can be used for the timely assessment of the severity of future influenza epidemics and pandemics. Timely pneumonia and influenza mortality reporting systems established in sentinel countries worldwide would help alert public health officials and allow prompt prevention and intervention strategies during future influenza epidemics and pandemics. PMID- 9209853 TI - Reproductive disorders among hairdressers. AB - To evaluate whether hairdressers have an increased risk of reproductive disorders, we conducted a historical cohort study in the Netherlands. Because exposure to reproduction toxic agents in hair salons may have changed over time, we studied two specific periods: conceptions in 1986-1988 and in 1991-1993. We ascertained 9,000 hairdressers and, as a comparison group, 9,000 clothing salesclerks from their respective trade associations. All were of reproductive age in the defined study periods. Frequency matching on 5-year age groups ensured comparability with regard to age. All women were approached by mail to complete a short, self-administered questionnaire on reproductive history, including questions on time-to-pregnancy, spontaneous abortion, livebirths, and congenital malformations. In the analyses, we used random effect models to account for correlated outcomes (multiple pregnancies per woman). The results show that hairdressers who conceived in 1986-1988 had an increased risk of prolonged time to-pregnancy of more than 12 months [odds ratio (OR) = 1.5; 95% confidence interval (CI) = 0.8-1.6], spontaneous abortion (OR = 1.6; 95% CI = 1.0-2.4), and a low-birthweight infant (OR = 1.5; 95% CI = 0.7-3.1). In both periods, more major malformations occurred among children of hairdressers, but numbers were small. These results indicate an increase in reproductive risks for hairdressers in earlier years that now seems to be disappearing. PMID- 9209854 TI - Dietary vitamin A intake in relation to child growth. AB - Severe deficits in ponderal and linear growth are problems of major public health significance among children in developing countries. We prospectively examined the association of dietary vitamin A intake with child growth among 28,740 Sudanese children ages 6-72 months. At baseline and at each 6-month visit, all subjects were weighed and measured. Dietary vitamin A intake during the prior 24 hours was assessed using recall of vitamin A-containing foods. Dietary vitamin A intake was associated with attained height and weight after controlling for age, sex, morbidity, and socioeconomic variables. Compared with children in the bottom quintile of intake, those in the top quintile were 11 mm taller [95% confidence interval (CI) = 8-13] and 237 gm heavier (95% CI = 153-320). Higher dietary vitamin A intake was also associated with reduced risk of stunting [relative risk (RR) for 5th vs 1st quintile = 0.7; 95% CI = 0.5-0.9] and wasting (RR = 0.7; 95% CI = 0.5-0.9). Adequate intake of foods containing vitamin A may improve child growth where vitamin A deficiency prevails, but this relation may not be due to vitamin A per se. PMID- 9209855 TI - Risk factors for work-related violent victimization. AB - This case-control study used the National Crime Victimization Survey database (a national sample of housing addresses) to examine sociodemographic risk factors for becoming a victim of work-related robbery and assault. Cases (N = 267) reported having been violently victimized in the previous 6 months. Controls (N = 1,783) were chosen from all nonvictims of violent crime at the end of the 6-month period. Risk factors varied by type of victimization, and differences were evident between men and women. Men less than 45 years of age had an increased risk for assault [odds ratio (OR) = 2.0-2.7], compared with those 55 years of age and older; and those with a family income of less than $40,000 had an increased risk for assault (OR = 1.7-1.9), compared with those having a family income of $50,000 or more. We found a decreased risk for those with a high school education (OR = 0.6), compared with those with some college education. For women, an increased risk was seen for ages 16-18 years (OR = 3.3) and 25-34 years (OR = 2.3), compared with those 55 years of age or older. Women who were divorced or separated (OR = 4.4) and never-married (OR = 2.1) were at higher risk than women who were married. We found a decreased risk for nonwhites (OR = 0.5), compared with whites. PMID- 9209856 TI - Environmental factors and the risk of salivary gland cancer. AB - Cancer of the major salivary glands is rare, and little is known about its etiology. We conducted a population-based case-control study to elucidate the risk factors for these tumors. Of 199 cases diagnosed with salivary gland tumors between 1989 and 1993, 150 (75%) were interviewed. We subsequently excluded nine cases based on review of pathology specimens. We identified 271 controls through random-digit dialing and the Health Care Finance Administration files; 191 (70%) were interviewed. Therapeutic medical radiation treatment to the head or neck [odds ratio (OR) = 2.6; 95% confidence interval (CI) = 0.84-8.1], full mouth dental x-rays (OR = 1.6; 95% CI = 1.0-2.7), and ultraviolet light treatment to the head or neck (OR = 1.9; 95% CI = 0.89-4.3) were associated with increased risk. These elevations in risk were largely limited to those exposed before 1955, when the exposure dose was substantially higher. Occupational exposure to radiation/radioactive materials (OR = 2.4; 95% CI = 1.0-5.4) and nickel compounds/alloys (OR = 6.0; 95% CI = 1.6-22.0), as well as employment in the rubber industry (OR = 7.0; 95% CI = 0.80-60.3), increased risk. In men, current smoking (OR = 2.1; 95% CI = 0.98-4.7) and heavy alcohol consumption (OR = 2.5; 95% CI = 1.1-5.7) were associated with risk, but these factors were not strongly related to salivary gland cancer in women. PMID- 9209857 TI - A prospective study of alcohol, smoking, caffeine, and the risk of duodenal ulcer in men. AB - The associations between smoking, caffeine, and alcohol intake and the risk of duodenal ulcer have rarely been investigated prospectively. We examined these associations in a prospective cohort of 47,806 men, 40-75 years of age, using a mailed baseline questionnaire in 1986, with follow-up every 2 years through 1992. During 6 years of follow-up, we documented 138 newly diagnosed cases of duodenal ulcer. After adjustment for age, energy-adjusted dietary fiber, body mass index, and use of aspirin or other nonsteroidal antiinflammatory drugs, current smoking was not associated with a substantial risk of duodenal ulcer [relative risk (RR) = 1.07; 95% confidence interval (CI) = 0.61-1.89]. Overall, past smokers were not at increased risk compared with never-smokers (RR = 0.99; 95% CI = 0.69-1.42). Adjusting for other risk factors, alcohol intake (comparing those who drink > 30 gm of alcohol per day to nondrinkers) was not associated with higher risk of duodenal ulcer (RR = 0.74; 95% CI = 0.42-1.29). We observed little association between caffeine, caffeine-containing beverages, and decaffeinated coffee and the risk of duodenal ulcer. These results indicate that smoking is not associated with a substantial increase in risk of duodenal ulcer, nor is high intake of alcohol and caffeine. PMID- 9209858 TI - Macronutrients, energy intake, and breast cancer risk: implications from different models. AB - We used data from a case-control study, conducted in Italy on 2,569 incident cases of breast cancer and 2,588 controls, to fit various energy adjustment models in the estimate of the effect of selected nutrients and alcohol on breast cancer risk. When we fit the standard multivariate and residual models, including total energy intake and the different macronutrients one at a time, the odds ratios (OR) related to 100 kcal per day were 0.90 for protein, 0.96 for saturated fat, 0.88 for unsaturated fat, 0.95 for sugar, 1.07 for starch, and 1.06 for alcohol. When we included all of the different sources of calorie intake in a single model with energy sources partitioned, the OR of adding 100 kcal per day was 0.91 for protein, 1.22 for saturated fat, 0.89 for unsaturated fat, 0.98 for sugar, 1.08 for starch, and 1.07 for alcohol. The estimates from the residual models that included various macronutrients separately were similar to those of the extended energy partition model. PMID- 9209859 TI - Controlling for continuous confounders in epidemiologic research. AB - Multiple regression models are commonly used to control for confounding in epidemiologic research. Parametric regression models, such as multiple logistic regression, are powerful tools to control for multiple covariates provided that the covariate-risk associations are correctly specified. Residual confounding may result, however, from inappropriate specification of the confounder-risk association. In this paper, we illustrate the order of magnitude of residual confounding that may occur with traditional approaches to control for continuous confounders in multiple logistic regression, such as inclusion of a single linear term or categorization of the confounder, under a variety of assumptions on the confounder-risk association. We show that inclusion of the confounder as a single linear term often provides satisfactory control for confounding even in situations in which the model assumptions are clearly violated. In contrast, categorization of the confounder may often lead to serious residual confounding if the number of categories is small. Alternative strategies to control for confounding, such as polynomial regression or linear spline regression, are a useful supplement to the more traditional approaches. PMID- 9209860 TI - Life-style factors and female infertility. AB - We summarize the epidemiologic literature on the effect of life-style factors such as cigarette smoking, alcohol and caffeine consumption, physical exercise, body mass index, and drug use on female infertility. We identified relevant papers through MEDLINE, Index Medicus, and a manual review of reference lists. Risk factors that affect the risk of primary tubal infertility and that were corroborated in two or more studies include use of intrauterine devices (especially the Dalkon Shield) and cigarette smoking. We identified extremes in body size as a risk factor for primary ovulatory infertility. Cocaine, marijuana and alcohol use, exercise, caffeine consumption, and ever-use of thyroid medications were possible risk factors for various subtypes of primary infertility. Few risk factors have been assessed or identified for secondary infertility or other less common subtypes, such as cervical or endometriosis related infertility. PMID- 9209861 TI - Sensitivity of the relation between cumulative magnetic field exposure and brain cancer mortality to choice of monitoring data grouping scheme. AB - We examined the effectiveness of alternative grouping strategies with respect to cumulative exposure to magnetic fields and brain cancer mortality among electric utility workers. We applied a statistically optimal job-exposure matrix to calculate cumulative exposure over full work histories. We studied the sensitivity of the exposure-disease relation by assigning an array of different quantitative exposure estimates based on six schemes for grouping exposure measurements. The quantitative relation between cumulative magnetic field exposure and brain cancer mortality appeared to be sensitive to the choice of grouping scheme, with the optimized grouping scheme indicating stronger relations than standard schemes. PMID- 9209862 TI - Antihypertensive drugs and fatal myocardial infarction in persons with uncomplicated hypertension. AB - We conducted a case-control study to evaluate the risk of fatal myocardial infarction in otherwise healthy treated hypertensive subjects according to the type of the antihypertensive drug used. The study encompassed 207 cases and 409 controls matched to cases on age, sex, and general practice. Compared with beta blocker users, the matched relative risk estimates for fatal myocardial infarction, adjusted for recent blood pressure, body mass index, smoking, duration of hypertension, and prior use of other antihypertensive drugs, were 0.7 [95% confidence interval (CI) = 0.4-1.2] for angiotensin-converting enzyme inhibitor users, 0.9 (95% CI = 0.5-1.5) for calcium channel blocker users, and 0.7 (95% CI = 0.4-1.2) for diuretic users. PMID- 9209863 TI - Routinely reported sexually transmitted diseases presage the evolution of the AIDS epidemic. AB - Because AIDS is largely transmitted through sexual intercourse, the descriptive epidemiology of sexually transmitted diseases of short incubation period could presage the incidence trend of AIDS after a time interval that approximates the incubation period of the disease. We have evaluated this hypothesis using data from the 50 states in the United States routinely reported to the Centers of Disease Control and Prevention from 1979 to 1994. We have estimated the dependence of the slope of the annual incidence rate of AIDS over the period 1987 1994 on the intercept and the slope of one or more of three common sexually transmitted diseases: gonorrhea, chancroid, and syphilis. When the parameters of two sexually transmitted diseases were used as predictor variables, the adjusted multiple correlation coefficient (Radj) ranged from 0.70 to 0.77. When the parameters of all three sexually transmitted diseases studied were used in the regression model, the Radj reached a high value of 0.79. We conclude that incidence data of three common sexually transmitted diseases during an 8-year period presage the evolution of the AIDS epidemic during the subsequent 8-year period. PMID- 9209864 TI - The importance of critically interpreting simulation studies. PMID- 9209866 TI - Public health professionals and interpersonal violence. PMID- 9209867 TI - Epidemiology, clinical science, and beyond. PMID- 9209868 TI - Epidemiology as a toolkit for clinical scientists. PMID- 9209869 TI - Epidemiology and the Internet. PMID- 9209870 TI - Epidemiology and the Internet. PMID- 9209871 TI - Ethylene oxide exposure and risk of spontaneous abortion, preterm birth, and postterm birth. PMID- 9209872 TI - Risk of cellular phone use still unaddressed empirically. PMID- 9209873 TI - Rib-vertebral angle asymmetry in idiopathic, neuromuscular and experimentally induced scoliosis. AB - The concave and convex rib-vertebral angle (RVA) at levels T2-T12 was measured on AP radiographs of 19 patients with right convex idiopathic thoracic scoliosis and 10 patients with major thoracic right convex neuromuscular scoliosis. The difference between the angles on the concave and the convex sides, the RVAD, was calculated. The RVAs were also measured on radiographs from three animal groups in which spinal curves had been induced experimentally in a variety of ways. Group 1 comprised 16 rabbits that had been subjected to selective electrostimulation of the latissimus dorsi, the erector spinae and the intercostal muscles. Group 2 comprised four dead rabbits whose spines had been subjected to manual bending. Group 3 comprised eight rabbits that had undergone mechanical elongation of one rib. In both the idiopathic and the neuromuscular group, the convex RVA was smaller than the concave RVA between levels T2 and T8, with a maximal difference between T4 to T5. From T9 to T12 the concave RVA was smaller than the convex. The RVA in relation to the scoliotic segment, i.e. the apex level of the curve and the two neighbouring vertebrae above and below this level, showed similar results. With increasing Cobb angle the RVADs increased linearly with the greatest difference at the second vertebra above the apex. In the three experimental groups the pattern of the RVADs between T6 to T12 was basically similar to the findings of the clinical study. From the results of these clinical and experimental studies, it is concluded that the typical pattern of the RVAs on the concave and convex sides seems to be independent of the underlying cause of the spinal curvature. It is likely that the RVADs result from a passive mechanical adaptation of the ribs to the lateral curvature of the spine. PMID- 9209875 TI - Neurophysiological mechanism of the unloading reflex as a prognostic factor in the early stages of idiopathic adolescent scoliosis. AB - This paper presents a new neurophysiological method of evaluating the risk of progression in idiopathic scoliosis in the early stages of the disease, by investigating the unloading reflex in paraspinal muscles. The study included 394 patients with scoliosis and 70 healthy children. Latency of the unloading reflex and repeatability of the rebound and silent period were analysed for each reflex evoked. Prolonged latency and minimal number of cycles are the most important factors in the evaluation of progressive idiopathic scoliosis. The method can be used to detect progressive idiopathic scoliosis in the early stages of the disease. The high sensitivity of the method means it can be used as a basis on which to take an early decision to perform surgery. PMID- 9209874 TI - Experimental determination of the effect of progressive sharp-angle spinal deformity on the spinal cord. AB - Neurological deficit is a serious though not well-known complication associated with spinal deformity. Sharp-angle kyphosis may be congenital, traumatic, degenerative, infectious, or iatrogenic in origin. Many kyphotic deformities are underestimated, thus leading to severe neurological deficit. In order to determine exactly what procedures of angulation the patients should undergo to stabilize the spine, which are major operations, the authors analyzed in an experimental model the effects of progressive sharp angulation on the anatomy of spinal canal and cord. We found that sharp anterior angulation of 50 degrees causes anterior-posterior stenosis and the dura will touch the spinal cord. At 90 degrees of angulation, the spinal cord will be squeezed and the pressure in the canal will be double what it was initially, probably leading to ischemia. The experimental confirmation (determination) of these angulations allows the physician in charge to define early in the treatment program when a surgical stabilization procedure should be included, before the angulation causes any neurological damage. PMID- 9209876 TI - Percutaneous lumbar discectomy in the treatment of lumbar discitis. AB - Lumbar disc infection, either after surgical discectomy or caused by haematogenous spread from other infection sources, is a severe complication. Specific antibiotic treatment has to be started as soon as possible to obtain satisfactory results in conservative treatment or operative fusion. The aim of this study was to analyse 16 cases of lumbar disc infection, treated with percutaneous lumbar discectomy (PLD) to obtain adequate amounts of tissue for histological examination and microbial culture. Between 1990 and 1994, 26 patients with vertebral osteomyelitis were treated. Sixteen patients, with an average age of 41.4 years (range 14-59 years), underwent a diagnostic PLD. Eight of them showed only moderate changes on computed tomograms (CT scans) and magnetic resonance (MR) images in the initial stages of the disease. The other eight showed more or less extensive osteolytic lesions of one or both vertebral bodies adjacent to the involved disc. The histology results showed non-specific discitis in nine patients and tuberculosis in one. In two patients an open biopsy had been performed, which showed non-specific discitis. Microbiological analysis revealed specific infection in 45% of the patients. These patients received a specific antibiotic treatment after antibiogram for an average of 33 days. Only three patients were treated surgically, with evacuation of the disc space and interbody fusion; the whole group received a spondylitis brace. All patients obtained satisfactory clinical results at the last follow-up regarding pain, mobility and spontaneous fusion of the involved disc space. In conclusion, PLD is a very helpful minimally invasive procedure in conservative treatment of lumbar discitis. PMID- 9209877 TI - A novel paraspinal surgical approach for lumbar lateral extraforaminal root entrapment. AB - We describe a new surgical route that we call the "crest approach" for treating extraforaminal disc herniation in the lumbar spine. This approach is useful only for the levels above L5-S1. It permits perfect root decompression without any bony resection that would contribute to instability. Muscle retraction and devascularization are reduced. Risk of nerve root lesions is minimal since the herniation is removed before root mobilization. Fifteen patients have been treated using this procedure. In all 15, pain and/or neurologic deficits remitted rapidly with no postoperative complications. In conclusion, the crest approach provides highly satisfactory operating conditions by simplifying exposure and greatly limiting the risk of complications. In our relatively limited experience using this procedure, only satisfactory results have been observed. PMID- 9209878 TI - The relationship between the magnetic resonance imaging appearance of the lumbar spine and low back pain, age and occupation in males. AB - The purpose of this study was to undertake a critical review of the potential role of magnetic resonance imaging (MRI) in the evaluation of low back pain (LBP) and to determine if there were differences in the MRI appearances between various occupational groups. The study group, 149 working men (78 aged 20-30 years and 71 aged 31-58 years) from five different occupations (car production workers, ambulance men, office staff, hospital porters and brewery draymen), underwent MRI of the lumbar spine. Thirty-four percent of the subjects had never experienced LBP. Twelve months later, the examination was repeated on 89 men. Age-related differences were seen in the MRI appearances of the lumbar spine. Disc degeneration was most common at L5/S1 and was significantly more prevalent (P < 0.01) in the older age group (52%) than in the younger age group (27%). Although LBP was more prevalent in the older subjects there was no relationship between LBP and disc degeneration. No differences in the MRI appearance of the lumbar spine were observed between the five occupational groups. Overall, 45% had 'abnormal' lumbar spines (evidence of disc degeneration, disc bulging or protrusion, facet hypertrophy, or nerve root compression). There was not a clear relationship between the MRI appearance of the lumbar spine and LBP. Thirty-two percent of asymptomatic subjects had 'abnormal' lumbar spines and 47% of all the subjects who had experienced LBP had 'normal' lumbar spines. During the 12-month follow-up period, 13 subjects experienced LBP for the first time. However, there was no change in the MRI appearances of their lumbar spines that could account for the onset of LBP. Although MRI is an excellent technique for evaluating the lumbar spine, this study shows that it does not provide a suitable pre-employment screening technique capable of identifying those at risk of LBP. PMID- 9209879 TI - Vertebral body MRI related to lumbar fusion results. AB - The evaluation of continued pain after a technically successful posterolateral lumbar spine fusion is often challenging. Although the intervertebral disc is often a source of low back pain, abnormal endplates may also be a focus of pain, and possibly a source of continued pain after a posterolateral fusion. MRI allows noninvasive evaluation for disc degeneration, as well as for abnormal endplates and adjacent vertebral body marrow. Previous studies have found inflammatory marrow changes, adjacent to abnormal endplates, associated with disc degeneration in low back pain patients. In this study, preoperative MRI scans in 89 posterolateral lumbar fusion patients were reviewed, by an independent radiologist, to determine whether vertebral body marrow changes adjacent to the endplates were related to continued pain. Independent chart review and follow-up telephone interview of all patients at a 4-year follow-up (mean) formed the basis for the clinical results. Vertebral body MRI signals consistent with inflammatory or fatty changes were found in 38% of patients, and always occurred adjacent to a degenerated disc. Inflammatory MRI vertebral body changes were significantly related to continued low back pain at P = 0.03. We conclude that posterolateral lumber fusion has a less predictable result for the subset of degenerative disc patients with abnormal endplates and associated marrow inflammation. More research is needed to determine the biological and biomechanical effects of posterolateral fusion upon the endplate within the fused segments. If indeed further study supports the hypothesis that abnormal endplates associated with inflammation are a source of pain, then treating the endplates directly by anterior fusion may be a preferred treatment for this subset of degenerative patients. PMID- 9209880 TI - Facet joint hypertrophy: the cross-sectional area of the superior articular process of L4 and L5. AB - With CT imaging, the lumbar facet joints are well visualised and enlargement secondary to degeneration may be noted. We measured the cross-sectional area of the superior articular process of the L5 facet joint in 100 consecutive CT scans and in 71 patients, the L4 process was also measured. We found that the mean cross-sectional area was significantly larger at L5 than at L4. Patient age and sex had no significant effect on the size at either L4 or L5. A review of the radiological reports revealed that the 13 patients with degenerative facet joints and radiologically normal discs did not have significantly larger facet joints than the 35 patients with disc disease and radiologically normal facet joints. In conclusion, the term "facet joint hypertrophy" should not be used when osteoarthritic changes are noted on CT scan, because these joints are not significantly larger than normal facet joints. PMID- 9209881 TI - Verification of the position of pedicle screws in lumbar spinal fusion. AB - Medial or lateral pedicle screw penetration with the potential to affect neural structures in a wellknown and frequent problem associated with posterior spinal fusion. We evaluated the placement of pedicle screws (n = 141) in 36 patients following posterior lumbar spinal fusion with Socon or Kluger instrumentation via a lateral transpedicular approach. The examination was based on CT and MR images performed after removal of the instrumentation, on average 1 year after implantation. We found seven pedicle screws with lateral cortical penetration of the pedicle and five screws with medial cortical penetration of the pedicle (8.5% pedicle penetration overall). No severe radicular complications accompanied these pedicle penetrations. The mean insertion angles of the pedicle screws at the L4 level were 22.6 degrees and 23.1 degrees for the left and the right side, respectively. At the L5 level the mean insertion angle was 20.5 degrees on the left side and 21.5 degrees on the right, and at the S1 level the mean angle was 16.2 degrees on the left and 15.2 degrees on the right. The results of this study indicate that the lateral transpedicular approach is a safe procedure for pedicle screw insertion. PMID- 9209882 TI - Load-displacement properties of the thoracolumbar calf spine: experimental results and comparison to known human data. AB - The availability of human cadaveric spine specimens for in vitro tests is limited and the risk of infection is now of vital concern. As an alternative or supplement, calf spines have been used as models for human spines, in particular to evaluate spinal implants. However, neither qualitative nor quantitative biomechanical data on calf spines are available for comparison with data on human specimens. The purpose of this study was to determine the fundamental biomechanical properties of calf spines and to compare them with existing data from human specimens. Range of motion, neutral zone, and stiffness properties of thoracolumbar calf spines (T6-L6) were determined under pure moment loading in flexion and extension, axial left/right rotation and right/left lateral bending. Biomechanical similarities were observed between the calf and reported human data, most notably in axial rotation and lateral bending. Range of motion in the lumbar spine in flexion and extension was somewhat less in the calf than that typically reported for the human, though still within the range. These results suggest that the calf spine can be considered on a limited basis as a model for the human spine in certain in vitro tests. PMID- 9209883 TI - Anterior cervical allograft arthrodesis and instrumentation: multilevel interbody grafting or strut graft reconstruction. AB - This retrospective study evaluated a single surgeon's series of patients treated by multilevel cervical disc excision (two or three levels), allograft tricortical iliac crest arthrodesis, and anterior instrumentation. The objective of this retrospective study was to compare fusion success and clinical outcome between multilevel Smith-Robinson interbody grafting and tricortical iliac strut graft reconstruction, both supplemented with anterior instrumentation in the cervical spine. The incidence of nonunion for cervical discectomy and fusion varies widely depending on the number of disc levels involved, type of bone graft used, and whether the anterior grafting is supplemented with instrumentation. An alternative to multilevel interbody fusion is corpectomy and strut grafting, in which the incidence of nonunion has been reported to be 27% with autograft and 41% with allograft. Sixty-four consecutive patients who underwent allograft tricortical iliac crest reconstruction and anterior cervical plating were studied. The average follow-up was 39 months. There were 38 patients in the discectomy and interbody grafting group and 26 patients in the corpectomy and strut graft reconstruction group. Pseudoarthrosis occurred in 42% of the anterior cervical interbody fusion patients and 31% of the corpectomy patients. Nonunion in two-level interbody fusions occurred in 36% of the patients as compared to 10% for patients with one-level corpectomies; while 54% of patients with three-level interbody fusions and 44% of patients with two-level corpectomies were noted to have pseudoarthrosis. Higher percentages of nonunion were noted in multilevel interbody grafting than in corpectomy with strut grafting and when more vertebral levels were involved. These radiographic and clinical findings underscore the shortcomings of multilevel anterior cervical allograft reconstruction with plating. Corpectomy may be the preferred method when multiple disc levels are fused. In addition, anterior corpectomy affords decompression of significant osteophytes in a safer and quicker manner. In retrospective studies, there is a need for long-term follow-up before accurate statements can be made about the study population. PMID- 9209885 TI - Spectrofluorimetric determination of 5-methoxypsoralen pharmacokinetic in patients' serum. AB - In medicine, psoriasis and vitiligo are most often treated with PUVA therapy (psoralen plus ultraviolet A). The determination of psoralen in patients' blood is necessary, as it is admitted that the therapeutic efficiency depends on drug concentration in patients' serum. The amount of UVA to administer is inversely proportional to serum peak concentration. High-performance liquid chromatography (HPLC) and gas chromatography are the most employed methods for determining psoralens in patients' serum. The 2 techniques are precise and very sensitive, but time consuming. The aim of this paper is to propose a suitable method which is rapid and simple. It is a spectrofluorimetric technique for 5-methoxypsoralen (5-MOP) determination in the serum of patients treated with PUVA therapy. 5-MOP extraction was carried out with an heptane/dichloromethane mixture (4/1; v/v), according to the Stolk method (1980). A calibration curve (CC) was plotted from 5 MOP concentrations (range 50, 100, 200, 300, 400, 500 ng/ml). The CC was linear with a good coefficient of correlation: r = 0.9971, and a suitable coefficient of variation (CV) of 7.0%. The recovery of the method ranged from 85.3 +/- 4.2 to 108 +/- 4.1%. The assay precision gave a CV ranging from 0.10 to 6.90%, with an error inferior to +/-10%. The method did not reveal any interference from serum components on the 5-MOP emission wavelength. The limit of detection of 5-MOP was 15 ng/ml. The proposed procedure was proved to be appropriate for a rapid determination of 5-MOP in patients' serum. This technique could also be employed for other psoralens used in PUVA therapy (e.g., 8-methoxypsoralen). PMID- 9209884 TI - Posterolateral lumbar fusion using facet joint fixation with biodegradable rods: a pilot study. AB - Roentgen stereophotogrammetric analysis (RSA) was used to assess whether there is a potential for biodegradable rods crossing the denuded facet joints to increase the stability and healing rate of lumbar posterolateral fusions. Eleven consecutive patients with lumbosacral disc/facet joint degeneration had a posterolateral fusion augmented with 2- or 3.2-mm biodegradable rods passing perpendicularly through the center of the denuded facet joints. The patients were followed-up with RSA in supine and erect positions monthly from the 2nd to the 6th postoperative month, and again 1 year postoperatively. All seven L5-S1 fusions healed. Four cases were stable as defined by RSA within 3 months, two within 6 months, and one within 1 year. One L4-S1 fusion could not be evaluated by RSA. None of the remaining three L4-S1 fusions fully healed. In all three cases 1- to 3-mm intervertebral translations remained at 1 year. None of the 11 fusions showed any radiographic signs of osteolysis around the biodegradable rods. The promising results of this pilot study indicate that posterolateral L5 S1 fusion augmented with transarticular biodegradable rods crossing the denuded facet joints may yield rapid intervertebral stabilization and a high healing rate without any adverse rod effects. This may be due to enhanced initial fusion stabilization and/or increased ossification induced by the rods. PMID- 9209886 TI - Expression of GD3 disialoganglioside antigen on peripheral T-lymphocytes in patients with disseminated malignant melanoma. AB - Disialoganglioside antigens GD2 and GD3 are expressed on most melanoma cells. On melanoma surrounding T-cells in immunohistological sections, disialogangliosides can also be found, as well as in a small % of T-lymphocytes in peripheral blood from healthy persons. In order to find out if there is a difference in ganglioside expression on peripheral T-lymphocytes between melanoma patients and healthy persons, we examined the expression of CD3 as T-lymphocytic antigen and GD2 or GD3 antigens, respectively, by flow cytometry. We used peripheral mononuclear blood cells of 12 patients with advanced disseminated malignant melanoma and of 12 healthy control donors. For immunostaining, murine monoclonal antibodies Leu-4, 14G2a and MB3.6 were used, recognizing CD3, GD2 and GD3. GD2 expression was found on only a low proportion of T-lymphocytes in patients and healthy persons (pat.: mean = 1.2% +/- 0.7%, co.: mean = 0.4% +/- 0.4%). Disialoganglioside antigen GD3, however, could be demonstrated on an average of 8.4% +/- 4.6% of patients' and on 4.0% +/- 2.1% of healthy persons' T-cells. There is a statistically significant difference (P < 0.01) between the data of patients' and control group. We conclude that there is a correlation between advanced malignant melanoma and expression of GD3 antigen on patients' peripheral T-lymphocytes. The immunological relevance of our findings is discussed. PMID- 9209887 TI - Identification of the LAMB3 hotspot mutation R635X in a Hungarian case of Herlitz junctional epidermolysis bullosa. AB - The Herlitz type of junctional epidermolysis bullosa (H-JEB) is a severe blistering disease affecting the skin and mucous membranes, which is usually lethal within the first year of life. The laminin 5 genes have been implicated as candidate genes for most patients with H-JEB. Recently, two hotspot mutations were delineated in the LAMB3 gene, known as R42X and R635X, and have been noted in over 50% of mutant LAMB3 alleles. Here, we present a case of H-JEB of Hungarian origin with a neonatal lethal outcome. Monoclonal antibody staining showed a lack of expression of the laminin 5 beta 3 chain, as a possible result of a mutation in one of the laminin 5 genes. Screening of the family identified the previously described mutation R635X in exon 14 of LAMB3 in each of the parents and one healthy sibling in the heterozygous form, while proband was homozygous for R635X, and the other sibling proved to be genotypically normal. These results underscore the widespread prevalence of R635X in H-JEB cases from around the world. PMID- 9209888 TI - Parallel intraindividual evaluation of the vasoconstrictory action and the anti allergic activity of topical corticosteroids. AB - The human skin blanching assay is a well established method for ranking the efficacy of corticosteroids after epicutaneous application. Vasoconstriction is a pharmacological activity, which correlates well with the clinical efficacy, the intensity of skin blanching after a single application under occlusion corresponding, generally, to the clinical efficacy after repeated application without occlusion. However, in studies dealing with the comparison between the vasoconstriction assay and the evaluation of the clinical effects on inflammatory skin diseases, some exceptions to this correlation have been reported. Therefore, in a pre-clinical phase, it would be useful to combine the blanching assay with at least one anti-inflammatory assay. In the present study the blanching assay and the allergic contact dermatitis inhibition test were performed in parallel, in order to compare the two testing procedures in the same group of subjects, utilizing standardized study designs supported by objective means of evaluation. Three commercial preparations of corticosteroids containing clobetasol propionate (CP), clobetasone butyrate (CB) and hydrocortisone acetate (HA), respectively, were employed both to treat nickel-induced positive patch test responses on the volar forearms, and to perform a vasoconstrictor assay on normal forearm skin in 16 nickel-sensitized healthy volunteers. For evaluating skin blanching, we employed colorimetric measurements, whereas for the quantitative determination of the inhibition of the intensity of allergic patch test reactions, 20 MHz B scanning supported by image analysis was used. Both colorimetric and echogenicity values enabled us to distinguish between the three corticosteroids (at the 17 h evaluation and the 64 h assessment, respectively). A fair correlation was noted between colorimetric and echogenicity values. Both testing procedures ranked the three corticosteroids in the expected order. Corticosteroid preparations should be compared using methods which allow different effects to be simultaneously monitored, without involving a high number of patients. We are proposing this double procedure for the parallel intraindividual evaluation of the vasoconstrictory action and the anti-allergic activity of topical steroids. PMID- 9209889 TI - Significant reduction of human monocyte chemotactic response to monocyte chemotactic protein 1 in patients with primary and metastatic malignant melanoma. AB - The recruitment of leukocytes from the peripheral blood is a key event for the development and composition of the inflammatory infiltrate in solid tumors and tumor metastases like malignant melanoma. Tumor-infiltrating lymphocytes (TIL) and tumor-associated macrophages (TAM) are thought to play a crucial role in tumor immunosurveillance. In malignant melanoma expression and secretion of monocyte-chemotactic protein 1 (MCP-1) have been demonstrated. MCP-1 serves as an attractant for monocytes and activated T-cells. In this study we addressed the question whether circulating monocytes show altered chemotaxis to MCP-1. Therefore the chemotactic responsiveness of monocytes towards MCP-1 was investigated in patients with primary and metastatic melanoma and compared to patients with basal cell carcinoma and healthy persons. The results show that monocytes from melanoma patients showed a significantly decreased chemotactic migration towards MCP-1 while chemotaxis to the stimulus N-formyl-methionyl leucyl-phenylalanine (FMLP) remained normal. Patients with basal cell carcinoma showed normal monocyte chemotaxis to all stimuli tested. In primary melanoma, there was no relation of the number of TAM or TIL to the decreased chemotaxis of circulating monocytes to MCP-1. From these data it can be concluded that circulating monocytes from patients with primary and metastatic melanoma show a MCP-1-specific decrease in chemotactic migration. This may be due to deactivation or modulation of the MCP-1-receptor expression on these cells. PMID- 9209890 TI - Expression pattern of human hair keratin basic 1 (hHb1) in hair follicle and pilomatricoma. AB - Using in situ hybridization, human hair keratin basic 1 (hHb1) gene expression was investigated in human normal scalp. hHb1 transcripts were specifically detected in the cortical cells of hair shaft but neither in the outer and inner root sheaths, nor in the hair cuticle or the medulla. hHb1 expression was detected strongly in cortical cells located from the beginning of the keratogenous zone up to the isthmus. These data specify the localization of hHb1 expression. Furthermore, neoplasms with follicular differentiation, including trichoblastoma, trichoepithelioma, pilomatricoma, pilar carcinoma and basal-cell carcinoma, were analysed for hHb1 gene expression. One of the 4 pilomatricoma specimens examined exhibited a very high level of hHb1 transcripts. Interestingly, this labeling was specifically associated to a transitional cell layer en route to trichocytic differentiation, providing evidence that in pilomatricoma, epithelial germ cells can differentiate towards hair shaft keratinocytes before evolving in ghost cells. PMID- 9209891 TI - Profile of cytokine mRNA expression in spontaneous and UV-induced skin lesions from actinic prurigo patients. AB - BACKGROUND: Actinic prurigo (AP) appears to be an immune-mediated disease triggered by exposure to ultraviolet light (UV). OBJECTIVE: To assess the profile of cytokine production in skin lesions from AP patients. METHODS: The cytokine production (IL-1 beta, IL-2, IL-4, IL-6, IL-10, IL-13, TNF-alpha, IFN-tau, and TGF-beta) in skin biopsies from 12 AP lesions was determined by a semiquantitative coupled reverse transcription-polymerase chain reaction. RESULTS: We found expression of TGF-beta and IL-13 genes in most AP skin lesions; IL-1 beta, IL-6, TNF-alpha, IFN-tau, and IL-10 were detected in some of these specimens. However, the levels of expression of all cytokines studied were not significantly different in AP skin lesions compared to nonlesional skin. CONCLUSIONS: TGF-beta and IL-13 might have a key role in both the inflammatory phenomenon and absence of significant expression of most cytokines in AP skin. The cytokine production in AP skin resembles that observed in rheumatoid synovium, a paucity in cytokine expression despite the presence of infiltrating activated mononuclear cells. PMID- 9209892 TI - Barrier disruption increases gene expression of cytokines and the 55 kD TNF receptor in murine skin. AB - The signalling mechanisms that regulate epidermal permeability barrier homeostasis are not known. Previous Northern blot analysis showed that both acute and chronic barrier disruption increase mRNA levels of several cytokines in murine epidermis. To further characterize the epidermal response to barrier abrogation, we used more sensitive, multi-probe RNase protection assays to measure the mRNA levels of additional cytokines, as well as cytokine receptors in acute and chronic models of barrier disruption. Normal mouse epidermis expressed interleukin (IL)-1 alpha, interferon-gamma (IFN-gamma), tumor necrosis factor alpha (TNF-alpha) and IL-6 mRNAs. Following tape-stripping, only the mRNA levels for TNF-alpha, IL-1 alpha, IL-1 beta and IL-6 increased at 2.5 and 7 h, and returned toward normal levels by 18 h. No mRNAs encoding TNF-beta, IL-2, IL-3, IL 4 or IL-5, were detected in the epidermis either under basal conditions or after tape-stripping. Similarly, in a chronic model, essential fatty acid deficiency, epidermal levels of TNF-alpha, IL-1 alpha, IL-1 beta and IL-6 mRNAs, but not IFN gamma mRNA, were elevated over controls; and again, mRNAs for the remaining probed cytokines were not detected. In contrast, in the dermis, only IL-1 beta mRNA levels increased 2.5 h after tape-stripping, and remained elevated at 18 h. mRNAs encoding the IL-1 (p60), IFN-gamma and IL-6 receptors were present in epidermis, but their levels remained unchanged following either acute or chronic barrier disruption. In contrast, epidermal TNF (p55) receptor mRNA levels were increased by 87% (P < 0.01) at 2.5 h, returned to control levels at 7 h and were increased by 68% (P < 0.03) at 18 h after tape-stripping. The increase at 2 h was confirmed by Northern blot analysis and was not prevented by latex occlusion performed immediately after tape-stripping mRNAs for the IL-1 (p80) receptor and TNF (p75) receptor were not detected in epidermis. Low levels of TNF (p55) receptor mRNA were present in the dermis, and they remained unchanged after tape stripping. The presence of specific receptor mRNAs in the epidermis and dermis suggests that these tissues are capable of responding in an autocrine and/or paracrine fashion to the cognate cytokines. These results suggest that epidermal cytokines produced after barrier disruption may initiate a cytokine cascade which could regulate cytokine and cytokine receptor production and/or inflammatory responses. PMID- 9209893 TI - Modulation of TGF-beta 1 production from human keratinocytes by UVB. AB - Transforming growth factor-beta (TGF-beta) plays an important role not only in cell growth control but also in inflammation and immunoregulation. There are at least five different isoforms of TGF-beta. TGF-beta 1 has a large variety of biological functions including the modulation of inflammation and the immune system and has most extensively been studied in skin. Since ultraviolet B (UVB) is known to induce skin erythema and immunosuppression, we sought to examine whether UVB would alter the expression and production of TGF-beta 1 in normal human keratinocytes. Using reverse transcription-polymerase chain reaction (RT PCR), constitutive expression of TGF-beta 1 mRNA was detected in keratinocytes and the level of TGF-beta 1 mRNA was increased 4 and 8 h after 300 J/m2 UVB irradiation. Production of TGF-beta 1 protein in culture supernatants assayed by ELISA was also increased at 24 h after irradiation. Cycloheximide treatment blocked this TGF-beta 1 protein induction indicating de novo protein synthesis of TGF-beta 1 from keratinocytes induced by UVB. These results suggest a possible role for TGF-beta 1 in UVB-induced skin inflammation and immunosuppression. PMID- 9209894 TI - Inhibitory effect of leukotrienes on luteinizing hormone release. AB - The aim of this work was to study the effect of high concentrations of leukotrienes on luteinizing hormone (LH) secretion in rat anterior pituitary cells. We also investigated the effect of leukotrienes in parallel with gonadotropin-releasing hormone (GnRH) action. Experiments were on cells gained from trypsinized pituitaries of female rats. Tests were performed by superfusion of the cells attached to cytodex-1 carrier beads. The LH content in samples of perfusate was estimated by radioimmunoassay. This work reports 48% inhibition of basic LH release by action of leukotriene C4 in superfused cells when applied continuously at a concentration of 100 nmol/l. Moreover, we have shown that leukotrienes suppressed GnRH-induced LH secretion in rat pituitary cells when applied in parallel to GnRH (1 nmol/l) as a 4-min pulse at a concentration of 0.1 nmol/l. GnRH-induced LH release was reduced to 66% of its value by leukotriene (LT) B4 (0.1 nmol/l) action; also to 54% by LTC4, 66% by LTD4 and 74% by LTE4 action. In contrast, arachidonic acid (50 pmol/l) and its other 5-lipoxygenase metabolites: 5-hydroperoxyeicosatetraenoic acid (5-HPETE) (50 pmol/l), or 5 hydroxyeicosatetraenoic acid (5-HETE) (50 pmol/l), had no inhibitory effect on GnRH-induced LH release. Arachidonic acid and 5-HETE potentiated GnRH-induced LH release up to 249% and 429%, respectively, when applied in parallel with GnRH (1 nmol/l) as a 4-min pulse at a concentration of 10 pmol/l. In our earlier work we have shown that several leukotrienes are potent stimulants of LH release. The present report documents the finding that the 5-lipoxygenase pathway is also involved in the inhibitory regulation of hormone release in anterior pituitary cells. PMID- 9209895 TI - Dopamine infusion enhances the adrenocorticotropic hormone and cortisol response to metoclopramide in hyperprolactinemic patients but not in normal subjects. AB - Based on the facts that prolactin and adrenocorticotropic hormone (ACTH) each seem to influence the secretion of the other, that dopamine is the established inhibitory factor for prolactin secretion and negatively modulates ACTH release, and finally that alterations of the central dopaminergic tone have been postulated in tumorous hyperprolactinemia, we studied the effects of pharmacological manipulations of the dopaminergic system on ACTH and cortisol secretion in patients bearing a prolactinoma and in normal subjects. Twenty-seven patients with a prolactin-secreting pituitary tumor and 12 healthy controls were submitted to three tests: (a) 4-h saline infusion; (b) 10 mg metoclopramide (MTC) as an intravenous bolus after a 2-h saline infusion; and (c) 4-h dopamine infusion at the dose of 0.01 microgram/kg/min with a 10-mg intravenous bolus of MTC given at the second hour of dopamine infusion. Administration of MTC, compared to saline, caused a moderate (not significant) plasma ACTH increase, and a significant cortisol increase (p < 0.05), both in hyperprolactinemic and normal subjects, without statistically significant differences between the two group. When MTC was administered during dopamine infusion, the ACTH and cortisol elevation was significantly potentiated in prolactinoma patients while it was similar in magnitude to that recorded after MTC alone in control subjects. These findings support the concept of an inhibitory role exerted by dopamine and are compatible with a stimulatory influence exerted by prolactin on corticotropin releasing hormone and ACTH secretion, and also favor the view of a reduced central dopaminergic tone in patients with tumorous hyperprolactinemia. PMID- 9209896 TI - Comparison between steroid hormones and cortisol in serum and follicular fluid in stimulated and unstimulated cycles of in vitro fertilization patients. AB - The present study was undertaken to evaluate the effects of the concentrations of serum and follicular fluid steroids and cortisol levels on the establishment of pregnancies in in vitro fertilization (IVF). Our study group consisted of 42 women (group A) who received gonadotropins for induction of ovulation for IVF. The control group included 23 women (group B) who underwent in vitro fertilization without stimulation. Serum estradiol and progesterone levels were significantly higher in group A than in group B. Serum and follicular fluid cortisol levels were similar in both groups A and B. There was no significant difference in the fertilization rates of the stimulated or unstimulated cycles. However, there were no pregnancies in group B whereas there was a 28.5% pregnancy rate in group A. There were no correlations between the estradiol, progesterone and cortisol levels when compared to the oocyte maturity and the fertilization rates in both groups of patients. PMID- 9209897 TI - Uterine leiomyomata and sterility: therapy with gonadotropin-releasing hormone agonists and leiomyomectomy. AB - The aim of this study was to obtain data about the pregnancy rate in patients with uterine leiomyomata after treatment with gonadotropin-releasing hormone (GnRH) agonists followed by myomectomy. Between 1987 and 1993, 61 patients with uterine leiomyomata and sterility underwent 6 months' GnRH agonist treatment, in part with a surgical intervention. Sixty-two per cent of the patients suffered from concomitant endometriosis. After hormonal therapy 41 patients underwent a myomectomy. According to sonographic and clinical criteria, there was no indication for the enucleation of the leiomyomata for the remaining 20 patients. Owing to the combined therapy, consisting of primary treatment of uterine leiomyomata with GnRH agonists, followed by surgical intervention, 25 patients (41%) suffering from long-term sterility (average 4 years) became pregnant. An early abortion occurred in only three cases (12%). No patient who underwent a myomectomy developed new myomata during the following pregnancy. Four patients suffering from a single leiomyoma became pregnant within the first 3 months after myomectomy, all of them conceiving spontaneously. Considering the high rate of spontaneous conceptions and the low abortion and complication rates during pregnancy, the combined therapy of GnRH agonists followed by myomectomy represents a major step forwards in the effective treatment of sterility in patients with uterine leiomyomata. PMID- 9209899 TI - A double-blind randomized study of the treatment of endometriosis with nafarelin or nafarelin plus norethisterone. AB - The objective of this study was to compare the efficacy of nafarelin 200 micrograms (Group A), nafarelin 400 micrograms (Group B) and the combination of nafarelin 200 micrograms and norethisterone 1.2 mg (Group C) daily, in treating symptoms of endometriosis, American Fertility Society score and adverse events during 6 months of treatment. A prospective, randomized, double-blind parallel group study was performed in two centers and 49 women with endometriosis diagnosed laparoscopically were included. The patients were seen monthly for physical examination and records were taken for bleeding pattern, symptom score and adverse events. A control laparoscopy was performed at the end of 6 months of treatment. All patients were followed 6 months after treatment. At 3 and 6 months the pelvic examination total score had decreased significantly in all three groups. The total endometriosis score was significantly reduced in Groups B and C. After 2 months the total symptom score showed a significant decrease in Groups B and C. The frequency of hot flushes during the first month of treatment was lowest in Group C, but during the rest of treatment there were no differences between the groups. Best bleeding control was obtained in Group C. We conclude that nafarelin 200 micrograms daily has as good an effect on endometriosis symptoms as nafarelin 400 micrograms daily, and the addition of norethisterone 1.2 mg results in fewer hot flushes and better bleeding control. PMID- 9209898 TI - Biochemical and histological effects of vaginal estriol and estradiol applications on the endometrium, myometrium and vagina of postmenopausal women. AB - Estrogen and progesterone receptors in the cytosol (ERc, PRc) and estrogen receptors in the nuclear compartment (ERn) were measured in the endometrium, myometrium and vagina of 29 postmenopausal women who underwent hysterectomy. The effects of vaginal estriol (0.5 mg daily) compared to 17 beta-estradiol (0.05 mg daily) therapy on these receptor levels were studied. In addition, the endometrium was examined by light microscopy for estrogenic stimulation. We found biochemical and histological signs of estrogenic stimulation in all three tissues after estradiol as well as estriol therapy. In the vagina the effect of both estrogens on the ERc concentration was different from that in the endometrium and myometrium. The effects of estradiol and estriol on the ERn were comparable in all three tissues. The PRc levels increased significantly in all tissues after estrogen therapy; in the myometrium it was significantly higher after estriol than after estradiol applications. In conclusion, there were no clear differences between vaginal estradiol and estriol medication with regard to the effects on receptor levels in vaginal and uterine tissues. In the histological studies at the light microscopy level similar signs of estrogen stimulation of the endometrium were found following estradiol and estriol medication. PMID- 9209900 TI - Does human chorionic gonadotropin have human thyrotropic activity in vivo? AB - Current evidence indicates that thyroid cells are sensitive to human chorionic gonadotropin (hCG) stimulation. In turn, thyroid hormones appear to influence ovarian and endometrial physiology and reproductive function. Our studies addressed the possible effect of endogenous and exogenous hCG on in vivo thyroid function in normal pregnancy and controlled ovarian hyperstimulation, respectively. Circulating concentrations of hCG in pregnant women during gestation were positively correlated with serum free thyroxine (r = 0.43, p = 0.02) and negatively correlated with thyrotropin levels in the same patients (r = 0.42, p = 0.02). By contrast, exogenous administration of hCG to effect follicular maturation in non-pregnant patients undergoing ovarian hyperstimulation resulted in lower circulating hCG concentrations than seen in pregnancy and failed to alter free thyroxine or thyrotropin levels (p > 0.22). Endogenous isoforms of hCG in early pregnancy appear to have thyrotropic activity in vivo. However, the results indicate that, under clinical conditions of controlled ovarian hyperstimulation for assisted reproduction, exogenous hCG does not affect the hypothalamic-pituitary-thyroid axis. PMID- 9209901 TI - An overview on the effectiveness of natural family planning. AB - Recent years have witnessed important developments in natural family planning (NFP), which is based on the observation of fertile and infertile periods of the menstrual cycle, so that the couple is able to know when sexual intercourse may lead to a pregnancy. A review of the main studies regarding the effectiveness of NFP showed a decrease in the Pearl Index and life table values from the early 1980s to date, indicating that progress both in the teaching and in the application of these contraception, methods has been achieved. The main cause of lack of success seems to be the misapplication of NFP rules, whereas the errors due to the method itself are few. Furthermore, it seems that the symptothermal method might give better results than the ovulation method, even though no comparative study has been carried out, and that the first studies on the lactational amenorrhea method show encouraging results. Finally, it seems that NFP is best suited for 'spacers' of pregnancies, rather than for 'limiters'. Indeed, the former are more likely to show good compliance, since the sexual abstinence periods are limited and an unwanted pregnancy is not regarded as a completely negative event. PMID- 9209902 TI - Chemoprevention of respiratory tract cancer. AB - Lung cancer is the leading cause of cancer death in the United States, and advances in therapy have accounted for an improvement in five-year survival in this disease from 9% to 13% over the last three decades. Molecular genetic evidence has confirmed the epidemiologic link between tobacco and lung cancer causation, and has clarified the etiology of the persistent risk of lung cancer development in former smokers. Retinoids have shown promise in aerodigestive cancer chemoprevention, both in the reversal of preneoplastic lesions and in the prevention of second primary cancers. After initial epidemiologic and dietary studies had linked beta-Carotene with cancer risk reduction, large randomized phase III studies of this compound have shown no evidence of benefit and some evidence of heightened lung cancer risk in active smokers on high dose supplemental beta-Carotene. Therefore, careful clinical, epidemiologic, and basic studies of retinoids using intermediate end point markers are necessary to determine the definitive role of these compounds in the chemoprevention of respiratory tract cancer, with a particular focus on former smokers. PMID- 9209903 TI - Prognostic markers in resectable non-small cell lung cancer. AB - Numerous prognostic factors have been identified in patients with resectable non small cell lung cancer (NSCLC) which may enable stratification of patients into subsets indicating risk of recurrence following complete resection. Such prognostic markers include a variety of clinico-pathologic factors such as tumor size, modal status, and histopathologic variables. Several serum tumor markers have also proven useful. Moreover, a wide variety of molecular markers have been described over the last decade, which can be classified as molecular genetic markers, metastatic propensity markers, differentiation markers, and proliferation markers. This article reviews those prognostic markers most likely to prove clinically useful from the perspective of guiding postresection treatment strategies in early stage NSCLC. PMID- 9209904 TI - Mediastinoscopy, thoracoscopy, and video-assisted thoracic surgery in the diagnosis and staging of lung cancer. AB - The surgical approach to the diagnosis and staging of lung cancer requires the assessment of the lung parenchyma, hilum, pleura, chest wall, and intrathoracic lymph nodes. Chest computerized tomography is sensitive in defining the location of the primary tumor, but is relatively insensitive to invasion. Similarly, radiographic imaging can identify lymph node enlargement, but lymph node enlargement alone is insufficient for accurate staging. To facilitate the tissue biopsies of both the primary tumor and potential sites of metastatic disease, video thoracoscopy has provided a useful complement to traditional bronchoscopy and mediastinoscopy. These instruments provide minimally invasive access to the lung, pleura, and ipsilateral lymph nodes. The combined application of thoracoscopy, bronchoscopy, and mediastinoscopy can provide intrathoracic staging information while minimizing surgical morbidity. PMID- 9209905 TI - Preoperative and postoperative care of standard and high risk surgical patients. AB - Much of the clinical teaching concerning preoperative evaluation is based upon clinical observations made several decades ago in patients undergoing thoracotomy for both benign and malignant diseases. More recent experience suggests that many "high risk" patients will tolerate pulmonary parenchymal resection. All patients being considered for surgery should have a complete history and physical examination, chest roentgenogram, and screening spirometry. If this initial evaluation reveals normal or mildly obstructed spirometry and absence of comorbid conditions, then the patient is at low risk and postoperative function may be accurately estimated by simple calculation. For patients with moderate or severe obstruction on spirometry (FEV1 less than 50% to 65% predicted), hilar disease, pleural disease, or prior surgery, quantitative radionuclide lung scanning is indicated to allow accurate calculation of postoperative function. For patients with a PPO FEV1 less than 0.8-1.0 L, additional risk stratification should be done after any preoperative interventions. Typically, this includes a formal or informal assessment of exercise capacity. Patients with severely impaired exercise capabilities are at very high risk for postoperative morbidity and mortality, and nonsurgical therapy should be considered. All active smokers at the time of evaluation should quit 3 to 4 weeks prior to planned surgery. Patients with purulent sputum should be treated with appropriate antibiotics. All patients with obstruction demonstrated on spirometry should be started on inhaled beta agonists, with or without inhaled corticosteroids. Postoperative management should focus on early mobilization. This requires adequate analgesia without excessive sedation. This is most easily achieved with local or regional analgesia techniques. The use of this approach, as well as patient-controlled analgesia, allows early mobilization and results in a short length of hospital stay. It should be recognized that if patients have an uncomplicated recovery and leave the hospital quickly (less than 6 days), postoperative pain will be a significant issue at home. Patients should be discharged with an adequate analgesia plan and an adequate supply of analgesic medications. PMID- 9209906 TI - Optimizing chemotherapy and radiotherapy in locally advanced non-small cell lung cancer. AB - In recent years, three treatment methods have been shown to improve overall survival of patients with locally advanced non-small cell lung cancer. These are: sequential combined radiotherapy and cisplatin-based chemotherapy, concurrent administration of cisplatin and thoracic radiotherapy, and accelerated hyper fractionated radiotherapy. Optimization of results can be attained by integrating these three treatment approaches. PMID- 9209907 TI - Chemotherapy for advanced non-small cell lung cancer. AB - Lung cancer is the most lethal cancer in both men and women. Given that chemotherapy for advanced disease is marginally beneficial and noncurative, its use must be governed judiciously, with each decision being evaluated individually for each patient. Chemotherapy for lung cancer has progressed over the past decade, and with the advent of new agents, its future looks promising. PMID- 9209908 TI - Future directions in lung cancer research and therapeutics. AB - Although immune and genetic approaches do not yet have substantial clinical exposure in NSCLC, it is hoped that the future will bring less toxic, more specific, and more effective methods to prevent the development of end-stage NSCLC. The principles of multitargeted and multimodality therapy to overcome tumor heterogeneity and resistance, and drug delivery issues have been developed over years of effort using chemotherapy. The lessons learned should not be forgotten, because they will remain applicable to newer biologic and genetic modalities. PMID- 9209909 TI - Lung cancer and chronic obstructive pulmonary disease. AB - The application of current knowledge and technology could dramatically improve the survival rate in both lung cancer and COPD, even before physicians and other health workers are finally able to convince the population that both personal and environmental tobacco smoke must be eliminated to begin to reduce the premature morbidity and mortality from lung cancer, airflow obstruction, and other smoking related diseases such as heart attack and stroke. PMID- 9209910 TI - Assessing "quality" in health care: issues in measurement and management. PMID- 9209911 TI - Quality of care initiatives in the French context. AB - The emergence of the issue of quality of health care systems of developed countries can be interpreted as the result of a growing pressure from payors and regulators on providers for increased accountability. As such, the form that this emergence takes is specific to each country's cultural, economic, and institutional context. A brief description of the French health care system and its political and functional dynamics is provided to set the stage and to explain how providers of care have responded to this pressure. The main emphasis has been put in a first period on the rationalization of medical practices using persuasive and coercive regulations to diffuse practice guidelines. As a consequence, medical data systems are developing in both the hospital and ambulatory care sectors. Quality assurance programmes are in their early years, and total quality management is still to come. PMID- 9209912 TI - Quality management in an academic integrated delivery system: the case of the University of Pennsylvania Health System. PMID- 9209913 TI - Toward quality improvement in a French hospital: structures and culture. AB - This paper describes the evolution of quality improvement efforts over the past decade in a 600-bed French teaching hospital, after reviewing some characteristics of the French hospital system. The sequential attempts to improve drug dispensation illustrate the evolution of quality improvement efforts. Initial studies conducted in voluntary medical wards led to the modification of traditional prescription forms. Resulting improvements were presented in Evaluation Committee meetings, inducing other wards, to undertake similar changes on their own. A more systemic program currently is underway. The success of initial efforts can be credited to the willingness of physicians and nurses to engage in changes when the need for change was evident in their daily work experience, in spite of the questionable validity of some of the original findings. Focusing on systemic processes rather than on individual faults allowed this involvement. Reassessing the validity of so-called medication errors also highlighted the importance of careful process analysis, which should be stressed during the development of quality assurance procedures. The variability inherent in clinical conditions calls for a flexible, outcome-oriented approach, as used in quality improvement methods. Quality improvement efforts are spreading throughout the hospital. However, diffusion of change from motivated work groups to the larger community remains a challenge. To obtain a consistent performance level across all parts of the organization, and to avoid losing impetus due to uncoordinated efforts, innovation must become a consistent feature of the hospital structure. PMID- 9209914 TI - Quality improvement in the US Veterans Health Administration. AB - The Veterans Health Administration, the largest government-operated health-care system in the United States, has been actively engaged in quality improvement activities since 1990. These activities have been implemented on both a system wide and facility-specific basis. Some quality improvement efforts have been targeted to specific clinical services; others relate to the overall process of providing patient care. This paper provides an overview of three quality improvement activities in the Veterans Health Administration and considers the research and managerial issues they raise. PMID- 9209915 TI - Beyond quality management methods: meeting the challenges of health care reform. AB - Over the last 20 years, the increasing complexity and technical intensiveness of health care in French hospitals have increased the level of uncertainty in the process of care. This paper argues that beyond quality management approaches, the most important issue in health care management is the need to implement new organizational methods in response to the dynamic changes that are transforming the care process. Uncertainty, complexity and speed can all be managed by standardizing operating procedures, but when quality management is applied to a complex system such as health care, a different approach is needed. One alternative to standardization can be found in new theories of organization that emphasize the flexibility of an organization, i.e. its capacity to adapt to uncertainty. Building on empirical work, this paper integrates these different theoretical perspectives and tries to provide insight into the kind of quality management methods that will allow hospitals to deal with the new constraints being placed on the process of care. PMID- 9209916 TI - The relationship between choice of outcome measure and hospital rank in general surgical procedures: implications for quality assessment. AB - OBJECTIVE: Institutional complication rates are often used to assess hospital quality of care, particularly for conditions and procedures where mortality rates are not useful because deaths are rare. The objective of this study was to assess the correlation among hospital quality assessment rankings based on adjusted mortality, complication and failure-to-rescue rates. DESIGN: This study used a clinically detailed administrative data set to compare severity and case-mix adjusted hospital outcome rankings for three different measures of quality of care: in-hospital death, complication and failure-to-rescue (in-hospital death following a complication). SETTING AND PATIENTS: Analysis of 74,647 patients who underwent general surgical procedures included in the 1991 and 1992 MedisGroups National Comparative Data Base. MEASUREMENTS: Adjusted outcomes of death, complication and failure to rescue based on multivariable logistic regression models. RESULTS: For 142 hospitals, the correlation between hospital rankings based on the death rate and those ranked by the complication rate was only 0.208 (P = 0.013). A similarly low correlation was present between the complication and failure rate rankings, r = -0.090 (P = 0.287). A higher correlation was observed between the death and failure rate rankings, r = 0.90 (P < 0.001). CONCLUSIONS: For general surgical procedures, hospital rank using the complication rate is poorly correlated with rankings using the death or failure rate. Complication rates should be used with great caution and should not be used in isolation when assessing hospital quality of care. PMID- 9209917 TI - Management tools and organization as key factors towards quality care: reflections from experience. AB - Health care organization in French hospitals has become an increasingly important issue, as efforts to ensure better cost control have increased financial constraints, as patients have demanded ever better results and quality, and as nurses' expectations for better working conditions have grown. Organizing a health care unit requires an articulation between individual efforts--necessary both for gathering accurate information on each patient and for providing patients with personalized care--as well as an integrated system of various logistics, specialized services such as nursing care, custodial care, technical examinations, and administrative procedures. Coordination between these different components continues to be a significant challenge to hospitals, as each category has developed independently its own specialty and sense of autonomy, resulting in different professional rationalities, cultures, approaches, and sometimes conflicting behaviour. Pointing out these difficulties is not sufficient to solve the problem, however, and since the management tools currently in use (activity measurements, procedures used to develop choices and judgements) are often kept in place for external reasons, they may actually perpetuate these behaviours. This paper is a set of reflections derived from a long period of experience and research in the management of French hospitals and health management institutions. It reports that the financial reforms implemented in budgeting procedures for French hospitals, and the efforts to control costs better in the national health insurance system, have resulted in new types of behaviour in physicians, nurses and health care managers, as well as the need for information that deals not only with health care activities but also with quality. PMID- 9209918 TI - The use of clinical guidelines to improve medical practice: main issues in the United States. AB - The use of clinical guidelines has become a key issue in the US health care system. In contrast to European systems, where such initiatives usually are controlled by one administrative agency, in the US there is a pluralistic approach and many kinds of guidelines coexist, initiated by health professions, managed care organizations, state or federal agencies, hospitals, and insurers. This paper reviews the main trends, indicating that guidelines will play an increasingly prominent role: use of institution-based guidelines vs national, professional, or state-based guidelines; use of more decision-support systems made possible by computerization and changes in cost containment strategies. Combining quality of care objectives with the business objectives of institutions increases the likelihood of a wider adoption by physicians. Several issues, such as the legal implications or the conflict of objectives, illustrate limits in the use of such standards to judge individual cases; however, most recent developments tend to reconcile individual decisions and what is known from probabilities on representative samples. By bringing such information into the decision process between physician and patient, the use of guidelines challenges the traditional asymmetry of information between professionals and patients. In a context of increasing health care costs, clinical guidelines represent a very useful tool for debating rationing issues and standard benefit packages, in order to make the system more equitable. Evaluations of the effectiveness of clinical guidelines on performance are contradictory, but when rigorous evaluations exist, clinical guidelines are found to be effective. The amount of improvement, however, may vary considerably. PMID- 9209919 TI - From clinical guidelines to quality assurance: the experience of Assistance Publique-Hopitaux de Paris. AB - OBJECTIVES: The objectives of this study are: to present the history of the development of clinical guidelines in France. to present the implementation of a program of clinical guidelines in Assistance Publique-Hopitaux de Paris (AP-HP), the regional public hospital system of the Paris metropolitan area, and to discuss the lessons learned from this program, which provide a basis for a better policy of guidelines at a national level. CONTENT: The objective of the AP-HP program is to promote the appropriate use of clinical guidelines and to evaluate their impact on quality of care and/or costs. Experiments such as controlled trials are used to test the effectiveness of some implementation strategies, to verify that guidelines do improve the process of care and to improve physician compliance with clinical guidelines. Guidelines are developed in a context of budget constraint, and the general aim of the program is to promote the more efficient use of available health resources. RESULTS: In a context of health care inflation, the desire of public authorities is to use guidelines as a tool for cost control strategy, either for ambulatory care or for hospitals. AP-HP experience shows that it is very difficult to develop a national program of clinical guidelines and to implement it in every hospital, as was done in France (but not clearly evaluated) in ambulatory care (under the name 'references medicales'). Guidelines need to be locally adapted, and the best implementation strategies depend on local resources, for example the existence of a pertinent information system. The role of financial incentives is present in French policies, and the role of the drug industry needs to be evaluated. PMID- 9209920 TI - Regional experience of evaluation of professional practice and quality assurance implementation in Aquitaine. AB - We report the establishment of a voluntary programme for developing the assessment of professional practice and quality assurance conducted since 1989 in the Aquitaine region of south-west France. This experience, in a region where there are 14,000 hospital beds in about a hundred institutions, has aimed at two objectives: (1) the adoption by the region's health professionals of medical guidelines for good practice and methods for measuring quality and outcomes obtained, by means of university training in evaluation and the conducting of several surveys of professional practice and clinical audits; (2) similar adoption by the regional political, administrative and medical directors of quality assurance procedures defined by an organizational audit concerning the prescription of preoperative tests in the university hospital and in the main public hospitals of the region. The critical impetus that this experience has created, together with other factors related to the political decision-makers and health managers of the region, has led to an operational regional structure for developing clinical evaluation and quality in Aquitaine. PMID- 9209921 TI - Another look at emergency room overcrowding: accessibility of the health services and quality of care. AB - PURPOSE: To describe both the social characteristics and the health needs of the medical users of a pediatric hospital Emergency Room with special emphasis on frequent use. STUDY SELECTION: Observational study on health services utilization and health care needs of young children consulting at a teaching hospital's Emergency Room. DATA SOURCES: Mother interview and medical record review. RESULTS OF DATA SYNTHESIS: Children from underprivileged strata are more often high Emergency Room users. Their preventive needs are satisfied but adequate follow-up of their medical problems is more often lacking. CONCLUSION: To understand why some achievable benefits are not achieved it is necessary to evaluate the varying performance of health services according to the social origin of the users. PMID- 9209922 TI - Renal replacement therapy for hemophiliacs. PMID- 9209923 TI - The pathogenesis of the diabetic kidney and the role of insulin-like growth factor. PMID- 9209924 TI - Serum albumin in peritoneal dialysis: clinical significance and important influences on its levels. PMID- 9209925 TI - Effects of pyruvate-based or lactate-based peritoneal dialysis solutions on neutrophil intracellular pH. AB - Acidic (pH 5.2) incubation mixtures containing pyruvate-based or lactate-based peritoneal dialysis solutions (PDSN's) induced comparable degrees of intracellular acidosis in neutrophils. However, addition of an acidic (pH 5.2), pyruvate-based PDSN to a pH-7.4, neutrophil/phosphate-buffered saline mixture brought about higher extracellular and intracellular (neutrophil) pH values when compared to the introduction of an equally acidic, lactate-based PDSN. This poor ability of acidic (pH 5.0-5.5), pyruvate-based PDSN's to resist alkalinizing influences is the cause for the above higher pH values. The higher intracellular pH levels so obtained may be a reason behind why acidic, pyruvate-based PDSN's appear to be more biocompatible than their equally acidic, lactate-based counterparts. PMID- 9209926 TI - Impact of spacing filaments external to hollow fibers on dialysate flow distribution and dialyzer performance. AB - A new type of dialyzer (PAN 650 SF Asahi) is analyzed in terms of hydraulic properties, solute clearances and dialysate flow distribution. The new type of dialyzer is a polyacrylonitrile hollow fiber filter, equipped with spacing filaments placed externally to the fibers to facilitate dialysate distribution and avoid channeling. The new filter is compared with a similar filter without spacing filaments. For this purpose, blood and dialysate side clearances have been measured in sequential dialysis session carried out randomly in the same patients. Furthermore, a last generation helical scanner (X-Press/HS1, Toshiba) has been utilized to analyze in vitro the flow distribution of dialysate inside the dialyzer. A contrast medium was injected and a sequence of images has been achieved on a longitudinal section of the dialyzer. This new method permits to avoid any bias due to the cylindrical shape of the dialyzer, since a 10 mm thick rectangular section is analyzed and not the entire body of the filter. The dialyzers equipped with spacing filaments displayed a significant improvement of the dialysate distribution as demonstrated by the radiological pattern. In detail, despite a channeling phenomenon in the peripherical region of the bundle is still present, this is remarkably reduced in comparison with the channelling phenomenon observed in the standard dialyzers. This improved distribution is confirmed by a significant improvement of the solute clearances. PMID- 9209927 TI - A new in vitro test method for calcification of bioprosthetic heart valves. AB - To investigate the calcification behavior of different bioprosthetic heart valves and verify possible hypotheses of the etiology of valve calcification, an accelerated pulse tester for bioprostheses was developed, whereby up to ten valves can be tested under identical test conditions. Each valve was mounted in a separate compartment on a piston and cyclically moved through a calcifying solution at frequencies of up to 800/min at 37 degrees C: An appropriate calcifying solution was evaluated by incubation tests of bovine and porcine tissue. Calcification was confirmed by measuring Ca and phosphate depletion by atomic absorption spectroscopy, von Kossa staining, EDAX, and microradiography. The first tests were successfully carried out on porcine valves that had been nondestructively assessed for tissue/stress anomalies by holographic interferometry prior to the calcification test. The tests showed that 75% of irregular fringe pattern areas corresponded to the calcification areas. PMID- 9209928 TI - Erythrothrombocytapheresis and plasmathrombocytapheresis with storage in T-sol of platelets collected by the new Amicus cell separator. AB - The Amicus cell separator is the latest apparatus introduced into the international market for high-yield, low WBC contamination and short-procedure time thrombocytapheresis. In its original configuration the apparatus collects platelets for subsequent resuspension in plasma and no collection of PRBC can be carried out along with thrombocytapheresis. In this paper we present the results of plasma-thrombocytapheresis and erythro-thrombocytapheresis after the adaptation of the Amicus to the collection and storage of platelets in a non plasma medium and the concurrent collection of PRBC. From our study it is concluded that PRBC collection doesn't modify the quality of the platelet product obtained from random donors (platelets pre-count 263x10e3/microliter) in terms of yield which is 4.6x10e11 platelets, WBC contamination (1.3x10e5) or procedure time (63 +/- 26 min.). The quality of the platelet products is satisfactory too, as measured by aggregation induced by collagen and ristocetin or by the substantial stability of membrane glycoproteins (CD62-63-51-36-42B). The concentrate had an average content of 58.8 g of hemoglobin per bag, a final volume of 376 +/- 13 ml, (after the addition of 100 ml of SAG-M) and a normal mechanic and osmotic fragility. PMID- 9209929 TI - Numerical modelling of blood flow behaviour in the valved catheter of the PUCA pump, a LVAD. AB - Mechanical heart assistance, performed by the PUlsatile CAtheter (PUCA) pump, chronologically takes place by sucking blood from the left ventricle and ejecting it into the ascending aorta. Within the pump activity the problem of hemolysis and clotting is encountered. In this study the influence of valve geometry on blood cell damage and stagnant zones has been investigated. A variable valve length coupled to a catheter ejection gap and a variable valve angle have been studied. In case of the studied valve, optimal parameter values have been determined. Compared to small catheter ejection gaps with a corresponding valve length, blood damage is found to be less for large ejection gaps with corresponding valve dimensions. In systole a valve positioned in a 0 degree angle proves to be best, whereas in diastole a +20 degree angle is preferable. Because the system is operating in both systole and diastole, a 0 degree angle valve is applied. PMID- 9209930 TI - Non-invasive blood glucose monitoring by means of near infrared spectroscopy: methods for improving the reliability of the calibration models. AB - The feasibility of using near infrared reflection spectroscopy for non-invasive blood glucose monitoring is discussed. Spectra were obtained using a diode-array spectrometer with a fiberoptic measuring head with a wavelength ranging from 800 nm to 1350 nm. Calibration was performed using partial least-squares regression and radial basis function networks. The results of different methods used to evaluate the quality of the recorded spectra in order to improve the reliability of the calibration models, are presented. PMID- 9209931 TI - Biomaterials for orthopedic surgery in osteoporotic bone: a comparative study in osteopenic rats. AB - To evaluate orthopedic devices in pathological bone, an experimental study was performed by implanting Titanium (Ti) and Hydroxyapatite (HA) rods in normal and osteopenic bone. Twenty-four rats were used: 12 were left intact ( CONTROL: C) while the other 12 were ovariectomized (OVX). After 4 months all the animals were submitted to the implant of Ti or HA in the left femoral condyle (Ti-C, HA-C, Ti OVX, HA-OVX). Two months later the animals were sacrificed for histomorphometric, ultrastructural and microanalytic studies. Our results show a significant difference between the Affinity Index (A.I.) of HA-C and Ti-C (77.0 +/- 7.4 vs 61.2 +/- 9.7) (p < 0.05). No significant differences were observed between the osteointegration of Ti-C and Ti-OVX (61.2 +/- 9.7 vs 48.2 +/- 6.7). Significant differences also exist between the osteointegration of HA-C and HA-OVX (77.0 +/- 7.4 vs 57.6 +/- 11.5) (p < 0.01). Microanalysis shows some modifications in Sulphur (S) concentration at the bone/biomaterial interface of the Ti-OVX group. Therefore our results confirmed the importance of biomaterials characteristics and of bone quality in osteointegration processes. PMID- 9209932 TI - Magnum wire for angioplasty of total and non-total coronary lesions. AB - The magnum wire is a stiff-shaft, blunt-tip wire constructed for recanalisation during angioplasty. Smaller series have demonstrated superior qualities compared to conventional wires. The purpose of this study was to analyze its feasibility in a larger number of procedures. A single centre database analysis identified the use of Magnum wire during 443 procedures accounting for 26% of all angioplasties from October 1992 to February 1995. There were 347 total occlusions and 222 non-total lesions. Technical success was 69% in total and 90% in non total lesions, whereas angiographic success was 58% and 75%, respectively. Stents were successfully implanted in 48 lesions; in 29 (8%) total occlusions and 19 (9%) non-total lesions. The total occlusion-related complication rate was 1.7%. Acute closure occurred in 4.1% of non-total lesions. In conclusion, Magnum wire is a feasible tool for angioplasty of total occlusions and severe stenoses. It is a cheap, simple and safe method for recanalisation. PMID- 9209933 TI - Troponin T, creatine kinase MB mass, and creatine kinase MB isoform ratio in the detection of myocardial damage during non-surgical coronary revascularization. AB - The presence of myocardial injury during non-surgical coronary revascularization has been evaluated by means of highly specific and sensitive biochemical markers. Troponin T, creatine kinase-MB isoenzyme mass concentration, and creatine kinase MB2/MB1 isoform ratio have been determined in 80 patients who underwent coronary revascularization with percutaneous transluminal coronary angioplasty (PTCA). Forty-five patients underwent balloon angioplasty, 15 rotational atherectomy, 10 directional atherectomy, and 10 elective coronary stenting. Serum concentration of the evaluated markers did not increase significantly after 57 uncomplicated revascularization procedures, including 15 rotablation procedures, nor after 8 PTCAs complicated by localized coronary type B and C dissections. Significant elevation of all markers above the upper limits of the reference interval (P < 0.05) was detected after occlusion of small side branches (< 0.5 mm diameter) in 5 patients. Creatine kinase MB2/MB1 isoform ratio was the earliest marker to increase. After recanalization of occluded vessels in 8/10 patients with 6-60 days old myocardial infarction only troponin T concentrations increased from a baseline of 0.28 microgram/l to a median peak of 0.80 microgram/l. This increase was statistically not significant (P = 0.12). In conclusion, myocardial damage was not detected following uncomplicated non-surgical revascularization obtained with different techniques. Markers of myocardial injury provide high sensitivity after small side branch occlusion. PMID- 9209934 TI - Guide wire outer coat shearing and embolisation: an unusual complication of pericardiocentesis. AB - We report a case of shearing of the outer coat of the guide wire and its embolization into the pulmonary artery during pericardiocentesis. This unusual foreign body was successfully removed by pervenous method. PMID- 9209935 TI - Incidence of congenital heart disease in Qatari children. AB - We surveyed the incidence of congenital heart disease in 49887 native live born children in the period between 1984 to 1994 in Qatar. Each child with clinically suspected congenital heart disease underwent echocardiographic examination. Magnetic resonance imaging, cardiac catheterization and surgical intervention were done at the discretion of the patient's pediatric cardiologist. Virtually no postmortem examinations were performed. Children with congenital heart disease were entered into a computerized database and were then followed for 1-11 years. Congenital heart disease was diagnosed in 610 of 49,887 children for an incidence of 12.23/1000 live births. The reasons for the high incidence were high proportion of small muscular ventricular septal defects discovered before the time of their spontaneous closure, referral to and follow up by a single group of pediatric cardiologists, location of the pediatric cardiology service in the same setting where nearly all of the deliveries took place, freely available health care service, and echocardiographic examination of every child with a clinical diagnosis of congenital heart disease. PMID- 9209936 TI - Delayed attainment of peak oxygen consumption after the end of exercise in patients with chronic heart failure. AB - Peak oxygen uptake (VO2) is attained at peak exercise in normal subjects. Recently, it was shown that the kinetics of the VO2 increase during exercise is slowed in chronic heart failure (CHF). We hypothesized that this may delay maximal VO2 after the end of exercise. We studied 21 patients who attained their peak VO2 15 s or more after cessation of a graded bicycle exercise test with breath-by-breath gas analysis (group 1). They were compared with 21 age- and sex matched CHF patients who did not do so (group 2) and 21 normal subjects (group 3). Peak VO2 occurred 30 +/- 10 s (15-45) after exercise and was 10 +/- 7% (3-31) higher than end-exercise VO2 (P < 0.001) in group 1. Group 1 patients had poorer functional status (NYHA class 3.0 +/- 0.2 vs. 2.4 +/- 0.5), a smaller ejection fraction (21 +/- 6 vs. 26 +/- 8%), a lower end-exercise VO2 (1156 +/- 251 vs. 1535 +/- 508 ml/min), a lower anaerobic threshold (762 +/- 183 vs. 970 +/- 265 ml/min), and an identical respiratory exchange ratio (1.09 +/- 0.13 vs. 1.06 +/- 0.12) relative to group 2 patients. The delta VO2/delta workrate ratio was lower (9.5 +/- 2.0 vs. 11.2 +/- 1.1 ml/W) and the half-time of VO2 recovery was longer (156 +/- 27 vs. 95 +/- 27 s) in group 1 than in group 2 (P < 0.05, P < 0.01 group 1 vs. group 2). Slow kinetics of the VO2 increase with exercise may delay achievement of peak VO2 beyond the maximal workrate achieved. Gas exchanges should thus be measured also during recovery so as not to underestimate the true peak VO2, especially in severe CHF patients referred for heart transplantation. PMID- 9209937 TI - Endothelin may be pathogenic in systemic sclerosis of the heart. AB - We evaluated 30 consecutive patients and 48 age- and sex-matched controls to explore the possibility of a pathogenic contribution by plasma endothelin-1 in the cardiac expression of systemic sclerosis. Venous plasma endothelin-1 was measured by radio-immunoassay and left ventricular function by echocardiography. The patient group had elevated plasma endothelin-1 (2.6 +/- 0.2 vs. 1.8 +/- 0.1 pmol/1, P < 0.001), but endothelin-1 was not related to age, heart rate, blood pressure, total peripheral resistance, disease duration or systemic sclerosis score. Endothelin-1 was related to left ventricular hypertrophy in terms of septal thickness (r = 0.33, P < 0.01) and left ventricular mass index (r = 0.32, P < 0.01). Plasma endothelin-1 was further related to measures indicating reduced left ventricular filling; left atrial emptying index (r = -0.50, P < 0.0005), the first third filling fraction (r = -0.31, P < 0.05) and the time velocity integral of Doppler early/late filling velocity (r = -0.40, P < 0.001). Furthermore, circulating endothelin-1 was related to impaired left ventricular contractility as estimated by pre-ejection period/left ventricular ejection time (r = 0.32, P < 0.01) and end-systolic wall stress/volume index (r = -0.30, P < 0.05). We conclude that plasma endothelin-1 is elevated in relation to the degree of left ventricular hypertrophy, diastolic dysfunction and impaired contractility in systemic sclerosis. It may be of pathogenic importance to the cardiac involvement in systemic sclerosis which is not mediated via an increase in systemic blood pressure. It is not yet clear whether our findings are exclusive to systemic sclerosis patients or represent a generalized phenomenon in patients with impaired left ventricular function. PMID- 9209938 TI - Clinical and echocardiographical study of the aortic homograft implantations in patients with Marfan syndrome. AB - The aim of the study was to assess the long-term results of surgical treatment with homogenic aortic grafts (HAGs) implantation in patients with Marfan syndrome. There were 31 patients with Marfan syndrome and aortic aneurysm who were operated on between 1980 and 1996. Aortic dissection was diagnosed in 14 patients, DeBakey Type I in six patients and Type II in eight patients. Four patients had to be operated urgently in cardiogenic shock with cardiac tamponade. Sealing up and reinforcement with strip of felt or Gore-Tex has been applied in 22 patients. The surgical modifications mentioned above have been applied since 1987 in all patients with the diameter of the aortic ring exceeding 30 mm or with active infective endocarditis or during reoperation. In 16 patients the space between the aortic homograft and patients own aortic wall was joined to the right atrial auricle. Patients were followed up for 12-179 months (average: 94.6 +/- 499). Three patients died in the early postoperative period and four patients died in the late postoperative period. Rethoracotomy because of bleeding complications was necessary in five patients. HAG damage was responsible for six other reoperations-new HAGs have been implanted in three patients and artificial prostheses were implanted in the other three patients. In the late follow-up period significant improvement in cardiac performance was observed in 24 patients (NYHA I or II). Survival probability of 15 years for the whole group was 80%. The lowest survival probability has been shown in the group of patients with DeBakey Type I aortic dissection (35% survived 15 years after operation). Echocardiographic follow-up has shown that the pressure gradient in HAG was low (7.4 +/- 6.2 mmHg). Only in two patients did the HAG gradient exceeded 20 mmHg. There were no significant differences concerning aortic ring diameters, dimensions of HAG and echocardiographic parameters between the group with surgical modifications, i.e. sealing up and reinforcement with strip of felt or Gore-Tex applied and the group in which these modifications were not applied. Homogenic aortic graft implantation as a method of surgical treatment of aortic aneurysm in patients with Marfan syndrome avoids postoperative anticoagulation, results in substantial improvement of cardiac performance and prolongs life. Surgical treatment should be considered in asymptomatic patients with large aneurysms (exceeding 55-65 mm) in patients with Marfan syndrome because there is a high risk of death in this group of patients in the case of dissection. PMID- 9209939 TI - Cardiac involvement is a constant finding in acute Chagas' disease: a clinical, parasitological and histopathological study. AB - During the last 8 years 58 acute cases of Chagas' disease were studied. Patients from an endemic area of the state of Barinas, Venezuela, showed fever (98%) and circulating forms of T. cruzi (100%), and were treated with oral benznidazole. The recorded mortality was 8.6%. Acute myocarditis was constantly found either in myocardial biopsies or at necropsy, even in patients without any other sign of cardiac compromise (36%), which was detected by chest X-ray in 58%, by 2D echocardiography in 52%, by resting ECG in 41% and by clinical findings in 27.5% of the patients. Cardiomegaly was due to pericardial effusion rather than ventricular dilatation in most instances. Treatment eliminated parasitemia but negativized serology in only 20% of patients. It also appeared to have little influence on the ongoing myocarditic process, emphasizing the need for better therapeutic schedules, able to avoid or control the early appearance of immunologic mechanisms and microcirculatory damage involved in the future development of chronic chagasic myocarditis. PMID- 9209940 TI - The specificity of exercise electrocardiography in women grouped by estrogen status. AB - We compared the specificity of exercise electrocardiography in 1880 men and 1818 women with women grouped by menopausal and estrogen replacement status. Specificity for > or = 1 mm horizontal or downsloping ST-segment depression was determined using angiography in 781 patients and using two other nonangiography based methods (a pretest probability-based method and a predictive accuracy-based method) in all patients. Using angiography, the specificities+/-SE were 84 +/- 2 for men, 79 +/- 3 for women, 81 +/- 5 for premenopausal women, 81 +/- 4 for postmenopausal women without estrogen replacement, and 77 +/- 5 for women on estrogen replacement. None of these were significantly different. For all patients, the respective specificities using the probability and predictive accuracy-based methods were 97 +/- 1 and 94 +/- 1 for men, 90 +/- 1 and 88 +/- 1 for women, 97 +/- 1 and 92 +/- 2 for premenopausal women, 92 +/- 4 and 88 +/- 3 for postmenopausal women without estrogen replacement, and 85 +/- 4 and 81 +/- 3 for women on estrogen replacement. (Men vs. all women groups except premenopausal women-P < 0.05). Therefore, the premenopausal women had significantly greater specificity than women on estrogen replacement (P < 0.001) and no difference in specificity with men. Women on estrogen replacement had a significantly lower specificity than postmenopausal women not on estrogen replacement (P < 0.05). These results suggest that estrogen replacement therapy and not naturally occurring estrogen has a role in producing false positive exercise electrocardiograms in women. PMID- 9209941 TI - Triggers of acute myocardial infarction regarding its site. AB - We have studied the incidence of possible triggers of the myocardial infarction regarding its site in 750 patients with anterior and 731 patients with inferior infarction. Infarctions occurred most frequently without recalling any triggering activity, especially in patients with anterior infarction (67 vs. 44%). Physical effort as the possible precipitator was also more frequent in anterior infarctions (22 vs. 16%). However, the onset of inferior infarction was more frequent during meteorological stress (9 vs. 2%), emotional stress (10 vs. 3%), after overeating (13 vs. 3%) and nicotine abuse (6 vs. 1.5%). These triggers were independent and highly significant (P < 0.02 in each case) discriminators of the site of myocardial infarction. Bimodal circadian rhythm, with primary peak between 6 and 9 h a.m. and the secondary peak between 3 and 6 p.m. was observed in patients which did not recall any triggering activity, and this was more pronounced in patients with inferior infarction. These results support the hypothesis that the influence of the vegetative tone is most pronounced in the onset of myocardial infarction of inferior wall. PMID- 9209942 TI - Serum calcium, phosphorus and albumin levels in relation to the angiographic severity of coronary artery disease. AB - Though calcium plays an important role in a number of biologic processes related to the pathogenesis of atherosclerosis, the relationship of serum calcium and phosphorus levels with the angiographic severity of coronary artery disease (CAD) is not known. We retrospectively studied 376 stable patients (age range 31-86 years, mean 59.2 +/- 10.5 years; 68% males) undergoing routine coronary angiography and related the angiographic severity of CAD with the serum levels of total and corrected calcium, phosphorus, albumin, total protein and bicarbonate. The primary variable studied was the number of vessels with haemodynamically significant disease. On univariate analysis, total serum calcium and serum albumin levels had a negative association with the number of vessels diseased (P = 0.046 and 0.057, respectively). Multiple regression analysis using age, sex, smoking, diabetes, hypertension, hyperlipidaemia, ethnicity and family history, in addition to serum calcium, phosphorus and albumin levels as the predictor variables, showed that serum albumin has an independent negative and serum phosphorus has an independent positive association with the angiographic severity of CAD (P = 0.04 and 0.003, respectively; n = 294). Serum phosphorus level also showed highly significant positive associations with the presence of total or subtotal occlusion and with most severe stenosis observed on angiography. A moderate change in the serum level of albumin or phosphorus confers a risk similar to that associated with smoking, as estimated by the odds ratios. PMID- 9209943 TI - Opposite effects of the remodeling of infarcted and non-infarcted myocardium on left ventricular function early after infarction in humans. An echocardiographic study in patients examined before and after myocardial infarction. AB - OBJECTIVE: The purpose of this study was to evaluate infarction-related changes in the infarcted and the non-infarcted myocardium using a baseline assessment of ventricular function obtained prior to the infarction. BACKGROUND: Experimental studies have shown that both infarcted and non-infarcted myocardium contribute to the process of left ventricular dilatation soon after the infarction, but no data exist on the effect that the infarct has on the pre-infarct ventricular morphology in humans. METHODS AND RESULTS: 10 patients, out of 721 admitted to our coronary care unit with a first acute myocardial infarction over a 3-year period, had had an echocardiographic examination performed before (354 +/- 407 days) and after (10 +/- 9 days) the infarction which were adequate for quantitative evaluation. Ventricular volume (Simpson) and regional wall motion (Centerline method) were evaluated by biplane apical sections and the endocardial length of the infarct and the non-infarct segments, imaged in a cross-sectional view at the papillary muscle level, were measured. After the infarction end diastolic and end-systolic ventricular volume increased (P = 0.0003 and P < 0.0001, respectively); diastolic and systolic infarct segment length increased (P = 0.011 and P = 0.0008, respectively), while non-infarct segment had only diastolic lengthening (P = 0.019), without systolic changes. The ejection fraction decreased after the infarction (P < 0.0001), in inverse relation to infarct size and in direct relation to diastolic non-infarct segment lengthening. In the five patients in whom there was a significant diastolic lengthening of non infarct segment (larger than mean +/- 2 S.D. of the interobserver variability) the decrease in ejection fraction was less than in the patients without significant lengthening of this segment (P = 0.017), despite a similar echocardiographic infarct size index. CONCLUSION: Ventricular enlargement early after myocardial infarction is due to both infarct expansion and lengthening of non-infarct segment. However, while systolic stretching of the infarct segment is a deleterious process that accounts for the increase in end-systolic volume, diastolic non-infarct segment lengthening is the expression of a functional compensatory mechanism that counteracts the reduction of the ventricular pump function secondary to the infarction. PMID- 9209945 TI - Acute Staphylococcal myocarditis masquerading as an acute myocardial infarction. AB - It is well known that viral myocarditis can mimic acute myocardial infarction and the differentiation of both diseases presents a diagnostic challenge. This report discusses the case presentation of an 18-year-old man who developed acute Staphylococcal myocarditis in the setting of acute tonsillitis in whom clinical, enzymatic, and electrocardiographic findings have led to tentative erroneous diagnosis of acute myocardial infarction. The outcome was favorable with cephalotin and captopril therapy. This is the first reported case of bacterial myocarditis masquerading as an acute myocardial infarction. PMID- 9209944 TI - Primary cardiac leiomyosarcoma: seven-year survival with combined surgical and adjuvant therapy. AB - Primary cardiac sarcomas constitute a rare entity that have been uniformly associated with poor long-term survival. A case of left atrial leiomyosarcoma involving the interatrial septum and the right atrial free wall and presenting with syncope and atrial fibrillation, is described. Two extensive surgical excisions followed by adjuvant radiation and chemotherapy improved survival with a good quality of life. This approach of combined surgical, medical and radiation therapy may offer better longterm outcome, since our patient is the longest survivor thus far reported. PMID- 9209946 TI - Congenital mitral stenosis: challenge of percutaneous transvenous mitral commissurotomy. AB - A 26-year-old woman with congenital mitral stenosis and embolic stroke was referred to our hospital. The echocardiogram showed a hypoplastic posterior mitral valve leaflet with short, unbalanced chordal attachments to the posteromedial papillary muscle. The mitral valve area was 0.9 cm2 by the pressure half-time method. There was no left atrial thrombus and spontaneous echo contrast. Percutaneous transvenous mitral commissurotomy was performed since the suggestion of surgical management was refused by her family members. A rupture at the chordae tendinae of the hypoplastic posterior papillary muscle developed during the procedure and needed mitral replacement. We advise that percutaneous transvenous mitral commissurotomy be avoided in adult patients with congenital mitral stenosis having an asymmetric and hypoplastic mitral valve. PMID- 9209947 TI - Congenital stenosis of isolated pulmonary vein: role of retrograde pulmonary vein catheterization. AB - We describe the technique of retrograde nontransseptal pulmonary vein catheterization in congenital isolated pulmonary vein stenosis, using a steerable left atrial cathelet. PMID- 9209949 TI - Aetiopathogenesis of peripartum cardiomyopathy: prolactin-selenium interaction? AB - The aetiology of peripartum cardiomyopathy is unknown. Fragmentary evidence from the published literature are synthesised to suggest a hypothesis that prolactin selenium interactions resulting in selenium deficiency and/or autoimmunity are responsible for peripartum cardiomyopathy. This hypothesis best explains the various known facts about the disease. The possible link between prolactin and selenium should be explored. PMID- 9209948 TI - Comments on 'A comparison of nifedipine once daily (Adalat LA), isosorbide mononitrate once daily and isosorbide dinitrate twice daily in patients with stable angina'. PMID- 9209950 TI - Progress report. A randomized multicenter European study comparing adjuvant radiotherapy, 6-mo chemotherapy, and combination therapy vs no-adjuvant treatment in resectable pancreatic cancer (ESPAC-1). AB - CONCLUSION: The ESPAC-1 trial is the largest study of its kind in pancreatic cancer and should definitively address the question of the role of conventional methods of adjuvant treatment in pancreatic cancer. BACKGROUND: At the joint International Association of Pancreatology and the European Pancreatic Club meeting in Mannheim, Germany (June 12-15, 1996) a satellite meeting of the European Study Group for Pancreatic Cancer (ESPAC) met to discuss the progress of the ESPAC-1 trial. METHODS: A randomized multicenter study to address which, if any, of the following adjuvant treatments are of benefit in patients with resectable pancreatic cancer: radiotherapy (40 Gy with 5-FU as a sensitizing agent), 6 mo of chemotherapy (5-FU and folinic acid), or a combination of these treatments. RESULTS: From February 1994 to June 1996 (the time of the Mannheim meeting) 221 patients so far have been recruited into the three treatment arms and one control arm. PMID- 9209951 TI - Urinary trypsinogen activation peptide (TAP) predicts severity in patients with acute pancreatitis. AB - CONCLUSIONS: Urinary TAP obtained within the first 48 h of the onset of symptoms can distinguish patients with severe acute pancreatitis. BACKGROUND: Urinary trypsinogen activation peptide (TAP) has recently been described as an early marker of severity in acute pancreatitis. METHODS: In a multicenter study, urine samples were collected for TAP concentration at 6-12, 24, and 48 h after admission from 139 patients with acute pancreatitis (99 with mild disease, 40 with severe disease) and from 50 control patients. Severity of acute pancreatitis was defined by the presence of organ failure and/ or pancreatic necrosis on dynamic contrast-enhanced computed tomography. RESULTS: Median urinary TAP in the 139 patients with acute pancreatitis compared to the 50 control patients was significantly higher at admission, 4.6 vs 0.8 ng/mL (p < 0.001), and 6-12 h, 1.9 vs 0.55 ng/mL (p = 0.04). Among patients who presented within 48 h of the onset of symptoms, the median urinary TAP for severe pancreatitis (9 patients) compared to mild pancreatitis (40 patients) was significantly higher at admission, 29.6 vs. 3.6 ng/mL (p = 0.001). Also, when obtained within 48 h of the onset of symptoms, all patients with severe pancreatitis had an admission urinary TAP level > 10 ng/mL. The sensitivity and specificity of an admission urinary TAP > or = 10 for severe pancreatitis was 100 and 85%, respectively. Given a cutoff of 10 ng/mL for an admission urinary TAP obtained within 48 h of the onset of symptoms, the negative predictive value was 100% for mild pancreatitis. PMID- 9209952 TI - Evaluation of UICC TNM classification for pancreatic cancer. A study of 228 patients. AB - CONCLUSION: A different stage grouping of TNM factors can improve the predictivity of the UICC TNM classification for pancreatic cancer. Nevertheless, the Japanese Pancreas Society (JPS) classification maintains a higher prognostic value. BACKGROUND: The use of a reliable staging classification facilitates the evaluation of anticancer treatments and the correct management of patients. The aim of the present study was to evaluate the prognostic value of three modified UICC TNM classifications, obtained by different stage grouping of the UICC TNM factors, comparing their predictivity to the standard UICC and the JPS classifications. METHODS: Clinical material consisted of 228 patients who underwent resection for pancreatic cancer. The reliability of the classifications was analyzed by the following methods: univariate analysis of stage survival curves; multivariate analysis of each classification after adjusting for grading and radicality; and correlation between the patients' distribution in the stages of each classification and in survival classes. RESULTS: The following modified UICC classification allowed a better differentiation of stage II and III survival (P = 0.08) than standard UICC (P = 0.74): stage I: T1N0M0; stage II: T1N1M0/T2N0M0: stage III: T2N1M0/T3 any NM0; stage IV: M1. The JPS classification better discriminated between the different stages (P < 0.001). All classifications had an independent prognostic value by multivariate analysis. The correlation between stages and survival classes was higher for the JPS classification than either UICC TNM classification or the modified UICC classifications. PMID- 9209953 TI - Pathological changes in pancreatic ducts from patients with chronic pancreatitis. AB - CONCLUSION: Chronic pancreatitis and restricted pancreatic outflow are accompanied by pathological changes in the ducts, including inflammation and alterations in the microvasculature. These changes and loss of epithelium provide a likely explanation for increased release of serum proteins, immunoglobulins, and lactoferrin into the juice, and the possibility of luminal contents entering the extracellular space and bloodstream. BACKGROUND: Enlargement of pancreatic ducts is a well-known phenomenon accompanying chronic pancreatitis and conditions restricting outflow of pancreatic juice. However, the relationship between ductal pathology and concomitant changes in the pancreatic juice is incompletely understood. METHODS: Segments of pancreatic ducts removed at surgery from patients with chronic pancreatitis and conditions restricting outflow were studied by light and electron microscopy to assess the pathological changes. RESULTS: Pathological changes in ducts from patients with chronic pancreatitis include chronic inflammation in the wall, enlarged and numerous capillaries packed with erythrocytes and leukocytes close to the lumen, and loss of epithelium and sometimes basement membrane. Plasma cells provide a source for increased immunoglobulins. Ducts from patients with diseases restricting outflow show significant pathology. PMID- 9209954 TI - Prevalence and clinical significance of combined K-ras mutation and p53 aberration in pancreatic adenocarcinoma. AB - CONCLUSION: This study could not attribute survival differences to the coincident acquisition of two common genetic alterations, K-ras mutation and p53 overexpression in pancreatic adenocarcinoma patients. Additionally, our data indicate the converse to be true: Those patients lacking both K-ras mutation and aberrant p53 expression showed the shortest survival when compared with cases showing either alteration or both. This study also showed the negative effect of K-ras mutation and p53 expression on pancreas cancer patients' survival after treatment with either radiation therapy or chemotherapy. BACKGROUND: Mutations of the oncogene K-ras at codon 12 are reported to be the most common genetic alteration in pancreatic carcinoma, whereas either overexpression or mutation of the tumor suppressor p53 gene is considered the most common genetic alteration in neoplasia of all types. p53 overexpression has been attributed to survival differences in pancreatic carcinoma, but such association is still controversial. No studies have fully documented the combined incidence of K-ras and p53 alterations in pancreatic adenocarcinoma, or their combined effect on patient survival in a large case series. The influence of radiation or chemotherapy in groups showing both, either, or neither mutation is also undocumented. METHODS: Paraffin-embedded tissue sections from 76 cases of pancreatic adenocarcinoma were cut for DNA extraction for K-ras analysis and immunohistochemical staining for aberrant p53 expression. K-ras mutation was determined by single-strand conformation polymorphism (SSCP) and slot-blot allele-specific oligonucleotide (ASO) hybridization of PCR-amplified DNA product p53 expression was scored on the basis of percent nuclear staining with the MAb DO7. RESULTS: Sixty-four of 76 cases (84%) showed K-ras mutation, p53 expression, or both, K-ras was mutated in 55 of 76 cases (72%). p53 was expressed in 33 of 76 cases (43%). Twenty-four of 76 cases (31%) showed both K-ras mutation and p53 expression. The presence of both alterations was not related to significant differences in tumor grade, stage, or survival compared to either alteration alone. A sizable subset (16% of cases) lacked either alteration, and surprisingly, this group showed the shortest median survival compared to those with K-ras mutation, p53 expression, or both (p = 0.024). Patients whose tumors were K-ras-negative showed the greatest difference in median survival with radiation therapy (median survival 30.8 mo vs 7.8 mo with no radiation, p = 0.005). PMID- 9209955 TI - The presence of the gallbladder is associated with the severity of acute biliary pancreatitis. AB - CONCLUSION: The presence of the gallbladder at the onset of acute biliary pancreatitis is associated with increased severity of the disease. One possible explanation is that gallbladder contraction might induce bile reflux into the pancreatic duct during the transfer of a gallstone through the ampulla. BACKGROUND: In clinical practice there is an impression that the presence of the gallbladder in patients with biliary pancreatitis may be associated with increased severity of the disease, compared to patients who have undergone cholecystectomy. METHODS: To test this hypothesis, we studied 266 cases with biliary pancreatitis. Patients were divided into two groups: (A) those who had a gallbladder in situ at the onset of biliary pancreatitis (n = 234, 88%) and (B) those who had undergone previous cholecystectomy (n = 32, 12%). RESULTS: Pancreatitis was more severe in group A than in group B, according to Glasgow criteria (> or = 3 positive, 66/210 = 31% vs 4/29 = 14%, p = 0.04); development of complications (77/234 = 33% vs 4/32 = 13%, p = 0.01); and mortality (40/234 = 17% vs 1/32 = 3%, p = 0.03). Furthermore, serum C-reactive protein levels on admission were over 150 mg/L twice as often in group A as in group B. PMID- 9209957 TI - Dissociated secretion of islet amyloid polypeptide and insulin in serum-free culture media conditioned by human pancreatic adenocarcinoma cell lines. AB - CONCLUSION: The cosecretion of insulin and islet amyloid polypeptide (IAPP) is altered when isolated rat pancreatic islets are incubated in culture media conditioned by human pancreatic cancer cells. BACKGROUND: Pancreatic cancer is usually associated with impaired glucose tolerance. This study investigates the tumor-derived influence on beta-cell secretion of pancreatic islets. METHODS: Four conditioned media were prepared from two human pancreatic cancer cell lines (Panc-1 and HPAF), a hamster pancreatic cancer cell line (PC-1), and a fibroblast cell line (Ag1523). Isolated rat pancreatic islets were incubated first in the conditioned media or nonconditioned control medium for 24 h, then in the same kind of media containing 100 microM carbamylcholine for 90 min. Insulin and IAPP secretion were measured by radioimmunoassay. RESULTS: Islets in media conditioned by Panc-1 and HPAF cells demonstrated dissociation of insulin and IAPP secretion. During 24-h incubation, the dissociation was expressed as selectively decreased insulin secretion. With addition of 100 microM carbamylcholine, the dissociation was expressed as normal secretion of insulin and hypersecretion of IAPP. As a result, the IAPP/insulin molar ratios were increased in both groups during both time periods. The islets in PC-1 and Ag1523 media did not show any significant changes in insulin and IAPP secretion. PMID- 9209956 TI - Acute pancreatitis in Sowetan Africans. A disease with high mortality and morbidity. AB - CONCLUSION: In African blacks, acute pancreatitis requiring hospital admission is a severe disease associated with a high mortality and significant long-term morbidity in surviving patients. BACKGROUND: It has been suggested that acute pancreatitis has a benign course in Africans in contrast to Western populations. The aim of the present study was to ascertain the incidence of acute pancreatitis at Baragwanath Hospital for a 1-yr period and to test the validity of the above hypothesis. METHODS: One hundred thirty-six patients with acute pancreatitis were retrospectively assessed. Fifty patients were available for a prospective follow up examination and underwent sonographic and biochemical investigations. Acute pancreatitis was diagnosed if the patient presented with the typical clinical picture and a raised serum amylase level > 800 U/L. RESULTS: The study consisted of 108 male and 28 female patients. Alcohol was identified as the predominant etiologic factor in 83.1%, biliary disease in 7.4%, and idiopathic causes in 6.6%. Substantial morbidity was encountered in 32.3% and was caused mainly by pancreatic complications, metabolic derangements, alcohol-related symptoms, and respiratory impairment. A portion (10.3%) of the patients developed further pancreatic pathology, such as pseudocysts, necroses, or an abscess. The overall mortality rate was 8.1%. Patients who died had a higher mean serum amylase, and most deaths occurred within 2 d of admission. Prospective follow-up after an average of 9.3 mo revealed serious morbidity in two-thirds of patients. Fifty-two percent suffered from severe abdominal pain, 36% complained of weight loss, and 18% were shown to have a sonographically abnormal pancreas. Fecal chymotrypsin levels indicated exocrine pancreatic impairment in 30.6%. PMID- 9209958 TI - Degeneration of intrapancreatic nerve fibers after chronic alcohol administration in mice. AB - CONCLUSION: These results provide morphological evidence for an alcohol-induced selective intrapancreatic nerve degeneration. This affected mainly the nerve fibers that are inhibitory of the exocrine pancreas, and might represent the morphological background of hypersecretory state of the pancreas in chronic alcoholism. METHODS: Intrapancreatic intrinsic nerves were studied by immunohistochemistry and electron microscopy after 4 mo of alcohol consumption and compared with control mice. RESULTS: A dense network of nerve fibers was observed in the normal mouse pancreas around the blood vessels and ending on the exocrine cells. The presence of VIP, NPY, PP, SP, and serotonin in these nerves was demonstrated by immunohistochemistry. Four months of alcohol consumption did not result in apparent morphological changes of the pancreas. However, the majority of periacinar nerve terminals showed degenerative changes. Synaptic vesicles were diminished in number in some other nerve processes, whereas the perivascular nerve fibers were relatively well preserved. A slight decrease was found in the intensity of VIP and SP immunoreactivity, and the PP fibers almost disappeared. PMID- 9209959 TI - Collection of pancreatic juice from growing pigs. A comparative study of the pouch method and the catheter method. AB - CONCLUSION: The results of this study demonstrate that there are large differences in the amount of pancreatic juice secreted and in the chemical and enzymatic composition of pancreatic juice when the pouch and the catheter methods were used, and these differences must be taken into consideration in future studies with either method. METHODS: A study was performed to compare the two most commonly used methods to collect pancreatic juice from growing pigs; namely, the pouch method (PM) and the catheter method (CM). In the first part of the study, three barrows (initial weight 37 kg) were fitted with a pancreatic pouch re-entrant cannula. An isolated pouch was prepared in which the pancreatic duct enters the duodenum. In the second part of the study, also with three barrows (initial weight 32 kg), a catheter was inserted into the pancreatic duct. RESULTS: At several points during the 24-h collection, the hourly rate of pancreatic juice secretion in CM pigs was larger (p < 0.05) than for PM pigs. CM pigs also had a higher (p < 0.05) daily volume of secretion, 4.09 vs 2.63 L/24 h for PM pigs. The pH of pancreatic juice collected from CM pigs was consistently higher (p < 0.01) throughout the 24-h collection. In contrast, the concentration and daily output of bicarbonate did not differ between CM and PM pigs. The concentration of protein in pancreatic juice from PM pigs (7.21 g/L) was higher (p < 0.001) than for CM pigs (4.08 g/L). Specific amylase and lipase and total amylase activities were greater (p < 0.01) in pancreatic juice collected from PM pigs. Specific and total carboxyl ester hydrolase and colipase activities were substantially (p < 0.01) larger in pancreatic juice collected from CM pigs. A major difference between the methods was that trypsin and chymotrypsin were fully active in pancreatic juice from PM pigs, whereas virtually no trypsin or chymotrypsin activity was detected in pancreatic juice from CM pigs. Specific and total chymotrypsin activities did not differ between PM and CM pigs. Both specific and total trypsin activities were substantially higher in pancreatic juice from CM pigs: 3682 U/L and 12,752 U/24 h, respectively, vs 1031 U/L and 2639 U/24 h, respectively, in pancreatic juice from PM pigs. PMID- 9209960 TI - Groove pancreatitis with recurrent duodenal obstruction. Report of a case successfully treated with pylorus-preserving pancreaticoduodenectomy. AB - Groove pancreatitis is a rare subtype of chronic pancreatitis that is difficult to distinguish from pancreatic carcinoma. Most reported patients have undergone a Whipple procedure because pancreatic cancer was not ruled out. We report a case of groove pancreatitis in a patient who presented with recurrent duodenal obstruction without biliary stricture. The diagnosis of groove pancreatitis was based on characteristic episodes of repeated duodenal obstruction and the absence of radiographic evidence of cancer. Subsequently, our patient underwent a successful pylorus-preserving pancreaticoduodenectomy (PPPD). PPPD is a favorable alternative to the Whipple operation for duodenal obstruction resulting from this disease. PMID- 9209961 TI - Low-energy transvenous cardioversion of atrial fibrillation using a single atrial lead system. AB - INTRODUCTION: Clinical studies have shown that electrical conversion of atrial fibrillation (AF) is feasible with transvenous catheter electrodes at low energies. We developed a single atrial lead system that allows atrial pacing, sensing, and defibrillation to improve and facilitate this new therapeutic option. METHODS AND RESULTS: The lead consists of a tripolar sensing, pacing, and defibrillation system. Two defibrillation coil electrodes are positioned on a stylet-guided lead. A ring electrode located between the two coils serves as the cathode for atrial sensing and pacing. We used this lead to cardiovert patients with acute or chronic AF. The distal coil was positioned in the coronary sinus, and the proximal coil and the ring electrode in the right atrium. R wave synchronized biphasic shocks were delivered between the two coils. Atrial signal detection and pacing were performed using the proximal coil and the ring electrode. Eight patients with acute AF (38 +/- 9 min) and eight patients with chronic AF (6.6 +/- 5 months) were included. The fluoroscopy time for lead placement was 3.5 +/- 4.3 minutes. The atrial defibrillation threshold was 2.0 +/ 1.4 J for patients with acute AF and 9.2 +/- 5.9 J for patients with chronic AF (P < 0.01). The signal amplitude detected was 1.7 +/- 1.1 mV during AF and 4.0 +/ 2.9 mV after restoration of sinus rhythm (P < 0.001). Atrial pacing was feasible at a threshold of 4.4 +/- 3.3 V (0.5-msec pulse width). CONCLUSIONS: Atrial signal detection, atrial pacing, and low-energy atrial defibrillation using this single atrial lead system is feasible in various clinical settings. This system might lead to a simpler, less invasive approach for internal atrial cardioversion. PMID- 9209962 TI - Magnetocardiography and 32-lead potential mapping: repolarization in normal subjects during pharmacologically induced stress. AB - Signals from 37 magnetocardiographic sensors and simultaneously recorded 32 ECG leads were obtained in three healthy male subjects (including two reinvestigations). After recordings at rest, the heart rate was increased by pharmacologic stress (117 to 142 beats/min). Comparison of the repolarization of rest and stress showed substantial changes in the magnetocardiogram (MCG) up to T wave inversions during stress. In the ECG only junctional ST-T segment shifts were present. For quantification, correlation coefficients between pairs of rest and stress MCG and rest and stress ECG distributions were calculated for the same time instant at the beginning of T wave under rest and stress conditions. In addition, equivalent electrical current dipole moment and magnetic dipole moment vectors were calculated from the MCG, and their change from rest to stress evaluated. Correlation coefficients for MCG comparison ranged from 0.3 to 0.5; ECG comparison suggested much less change from stress, ranging from 0.7 to 1.0. Current dipole moment changes at T wave onset were marginal; in contrast, the magnetic dipole moment changed substantially. Since the magnetic dipole reflects vortex currents, changes in its intensity and/or orientation during repolarization suggest this as the biophysical basis of the striking difference in the response of the MCG and ECG to pharmacologic stress. Normal ECG findings at rest and under stress in healthy subjects support the conclusion that the repolarization changes in the MCG were of nonpathologic origin. PMID- 9209963 TI - Effects of slow pathway ablation on fast pathway function in patients with atrioventricular nodal reentrant tachycardia. AB - INTRODUCTION: This study investigated whether fast pathway conduction properties are altered by slow pathway ablation in patients with AV nodal reentrant tachycardia. METHODS AND RESULTS: Forty consecutive patients who underwent successful ablation of the slow pathway were prospective subjects for the study. Isoproterenol was used to enhance conduction and to differentiate interactive mechanisms. Potential electrotonic interactions were assessed by comparing patients with and those without residual dual AV node physiology after slow pathway ablation. Paired and unpaired t-tests were used when appropriate P < 0.05 was considered statistically significant. In the entire study population, heart rates were not significantly different before and after slow pathway ablation (RR = 770 +/- 114 msec before and 745 +/- 99 msec after, P = 0.07). Anterograde fast pathway conduction properties were unchanged after slow pathway ablation (effective refractory period, 348 +/- 84 msec before and 336 +/- 86 msec after, P = 0.13; shortest 1:1 conduction, 410 +/- 93 msec before and 400 +/- 82 msec after, P = 0.39). Retrograde fast pathway characteristics also were similar before and after ablation. Neither anterograde nor retrograde fast pathway conduction properties during isoproterenol infusion were changed by slow pathway ablation. When the study population was further divided into patients with (n = 13) or without (n = 27) residual dual AV node physiology, no significant change was detected in fast pathway function in either group after slow pathway ablation. CONCLUSIONS: Fast pathway conduction characteristics were not affected by slow pathway ablation. In patients with AV nodal reentrant tachycardia, observations suggest that fast and slow pathways are functionally distinct. PMID- 9209964 TI - Evaluation of fast pathway function: the importance of autonomic tone. PMID- 9209965 TI - QT and JT dispersion in children with long QT syndrome. AB - INTRODUCTION: Abnormalities of ventricular repolarization leading to ventricular arrhythmias place children with long QT syndrome at high risk for sudden death. Dispersion of the QT (QTd) and JT (JTd) intervals, as markers of cardiac electrical heterogeneity, may be helpful in evaluating children with long QT syndrome and identifying a subset of patients at high risk for development of critical ventricular arrhythmias (ventricular tachycardia, torsades de pointes, and/or cardiac arrest). METHODS AND RESULTS: The QTd and JTd intervals in 39 children with long QT syndrome were compared to those of 50 normal age-matched children. In the long QT syndrome group, QTd measured 81 +/- 70 msec compared to 28 +/- 14 msec in the control group (P < 0.05), and JTd in the long QT syndrome group was 80 +/- 69 msec compared to 25 +/- 15 msec in the control group (P < 0.05). CONCLUSION: Children with long QT syndrome have an increased QTd and JTd when compared to normal controls. A QTd or JTd > or = 55 msec correlates with the presence of critical ventricular arrhythmias. These ECG measures of dispersion can be useful in stratifying children with the long QT syndrome who are at higher risk for developing critical ventricular arrhythmias. PMID- 9209966 TI - Optimized first phase tilt in "parallel-series" biphasic waveform. AB - INTRODUCTION: A biphasic defibrillation waveform can achieve a large second phase leading-edge voltage by a "parallel-series" switching system. Recently, such a system using two 30-microF capacitances demonstrated better defibrillation threshold than standard waveforms available in current implantable devices. However, the optimized tilt of such a "parallel-series" system had not been defined. METHODS AND RESULTS: Defibrillation thresholds were evaluated for five different biphasic "parallel-series" waveforms (60/15 microF) and a biphasic "parallel-parallel" waveform (60/60 microF) in 12 anesthetized pigs. The five "parallel-series" waveforms had first phase tilts of 40%, 50%, 60%, 70%, and 80% with second phase pulse width of 3 msec. The "parallel-parallel" waveform had first phase tilt of 50% with second phase pulse width of 3 msec. The defibrillation lead system comprised a left pectoral "hot can" electrode (cathode) and a right ventricular lead (anode). The stored energy at defibrillation threshold of the "parallel-series" waveform with first phase tilts of 40%, 50%, 60%, 70%, and 80% was 7.0 +/- 2.1, 6.1 +/- 2.8, 6.8 +/- 2.8, 7.2 +/- 2.9, and 8.4 +/- 3.1 J, respectively. The stored energy of the "parallel-series" waveform with a 50% first phase tilt was 16% less than the nonswitching "parallel parallel" waveform (7.3 +/- 2.8 J, P = 0.006). CONCLUSIONS: A first phase tilt of 50% maximized defibrillation efficacy of biphasic waveforms implemented with a "parallel-series" switching system. This optimized "parallel-series" waveform was more efficient than the comparable "parallel-parallel" biphasic waveform having the same first phase capacitance and tilt. PMID- 9209967 TI - Responses to norepinephrine of normal and "ischemic" canine Purkinje fibers are consistent with activation of different alpha 1-receptor subtypes. AB - INTRODUCTION: Previously we found that WB4101 (WB) 10(-7) M competitively blocks three alpha 1-adrenergic receptor-effector responses: the increase in normal automaticity occurring in Purkinje fibers (PF) at high membrane potentials; the increase in abnormal automaticity occurring in PF at depolarized membrane potentials; and the prolongation of PF action potential duration. These observations are consistent with two different hypotheses: (1) WB blocks a single alpha 1-receptor subtype, which subserves different effector pathways; and (2) WB blocks different receptor subtypes, each of which subserves an independent pathway. The aim of this study was to test both hypotheses. METHODS AND RESULTS: We used standard microelectrode techniques to study the concentration-dependent actions of three alpha 1-adrenoreceptor blockers (WB [alpha 1A > or = alpha 1D], 5-methylurapidil [5-MU] [alpha 1A > > alpha 1D], and UK52,046 [nonselective]) or norepinephrine (NE) effects in normal PF and in PF depolarized with a simulated ischemic solution ([K+]o = 10 mM; pO2 < 20 mmHg; pH 6.8; maximum diastolic potential -60 +/- 1 mV). In normally polarized PF, concentration-dependent actions of all blockers on both the positive chronotropic response and the prolongation of action potential duration completely coincide. In contrast, the response to NE of abnormal automaticity in "ischemic" PF differs from normals: there is a high sensitivity to WB and 5-MU and no response to UK52,046. CONCLUSIONS: (1) A single receptor subtype appears responsible for both the alpha 1-induced prolongation of repolarization and the positive chronotropic effect in normal PF. (2) Two different receptor subtypes may be responsible for the alpha 1 induced effects on automaticity in normal and ischemic fibers. It is likely that the latter one is alpha 1A, and that consideration of antiarrhythmic therapy with alpha 1-adrenergic blockers should focus on this subtype as a potential target. PMID- 9209969 TI - Electrophysiologic effects of interactions between activated canine neutrophils and cardiac myocytes. AB - INTRODUCTION: Myocardial ischemia causes neutrophils to bind to activated myocytes and liberate platelet-activating factor (PAF). PAF causes delayed repolarization, early afterdepolarizations (EADs), and arrest of repolarization. We studied the effect of activation of neutrophils bound to canine cardiac myocytes to determine if such activation causes PAF generation and similar changes in transmembrane potentials. METHODS AND RESULTS: Myocytes from canine left ventricle and neutrophils from the same dog were superfused with Tyrode's solution and transmembrane potentials recorded from the former. Neutrophils (100 microL, 10(6)/mL) were added and allowed to bind to the myocytes. Neutrophils were activated with 1% zymosan-activated serum (ZAS). CV-6209 (100 nM) was used to block receptors for PAF. Liberation of PAF by activated neutrophils was quantified with a commercial radioimmunoassay kit. Neutrophils activated with ZAS caused changes in myocyte transmembrane potentials like those induced by PAF: action potential prolongation, runs of EAD, and periods of plateau arrest. PAF receptor blockade prevented neutrophil activation from altering transmembrane potentials. Neutrophils activated with 1% ZAS liberated significant amounts of PAF. CONCLUSIONS: When neutrophils bound to cardiac myocytes are activated by exposure to 1% ZAS, they cause prompt and consistent changes in myocyte electrical activity that could be arrhythmogenic for the in situ heart. These changes are similar to those caused by PAF in pharmacologic studies. Neutrophils activated in this manner generate PAF, and the effects of their activation are prevented by blockade of PAF receptors. We conclude that, during reperfusion of ischemic myocardium, PAF generated by activated neutrophils most likely is a cause of some arrhythmias. PMID- 9209970 TI - Bundle branch reentry ventricular tachycardia with two distinct left bundle branch block morphologies. AB - INTRODUCTION: Bundle branch reentry ventricular tachycardia (VT) is usually amenable to treatment with radiofrequency ablation. Different QRS morphologies during VT are possible when anterograde ventricular activation is over the left bundle branch. Manifestations of this reentrant tachycardia with more than one QRS morphology with anterograde activation via the right bundle have not been reported and would be unusual due to the more discrete anatomy of the right bundle branch. METHODS AND RESULTS: An electrophysiologic study was conducted in a patient with dilated ventricle and diminished ventricular function with VT. Typical characteristics of bundle branch reentry were noted when VT was induced. The study was notable for the presence of a right bundle recording only during macroreentrant beats or VT and the distal location of the recording. Radiofrequency ablation was performed. Postablation stimulation again induced VT, proven to be of the same bundle branch reentry mechanism but of a different QRS morphology. A second ablation was required for complete ablation of this patient's bundle branch reentry VT. CONCLUSION: In bundle branch reentry utilizing the left bundle as the retrograde limb and the right bundle branch as the anterograde limb of the circuit, VT of more than one distinct morphology can be seen. Careful evaluation to assess for the persistence of VT of the same mechanism is necessary to ensure complete ablation of the reentrant circuit. Preexisting right bundle disease and a dilated heart with more dispersed distal right bundle branches may predispose to such a phenomenon. PMID- 9209968 TI - Early afterdepolarizations produced by d,l-sotalol and clofilium. AB - INTRODUCTION: The roles for L-type calcium current and Na-Ca exchange in early afterdepolarizations (EADs) attending d,l-sotalol and clofilium were examined in canine Purkinje fibers and in enzymatically dispersed myocytes from canine subepicardium. METHODS AND RESULTS: Spontaneous EADs were compared to EAD formation potentiated by stimulation of Na-Ca exchange and facilitation of ICa-L (Bay K8644). Bay K8644 (10(-8) M) and stimulation of Na-Ca exchange potentiated bradycardia-dependent EADs. Stimulation of Na-Ca exchange in Purkinje fibers pretreated with d,l-sotalol (10(-5) M) and clofilium (10(-7) M) induced EADs at takeoff potentials negative (-63 +/- 4 and -62 +/- 4 mV, respectively) to EADs potentiated by Bay K8644 (10(-8) M) (-33 +/- 2 and -34 +/- 2 mV, respectively, P < 0.05), or EADs induced by Bay K8644 alone (10(-6) M) (-31 +/- 5 mV). In myocytes, Bay K8644 (10(-8) M) potentiated EADs in d,l-sotalol- (10(-6) to 10(-4) M) or clofilium-treated (10(-9) to 10(-7) M) cells at reduced potentials (-10 +/- 3 and -10 +/- 4 mV, respectively) compared to EADs elicited by clofilium or d,l sotalol alone (-25 +/- 3 and -24 +/- 3 mV, respectively), or stimulation of Na-Ca exchange in the presence of d,l-sotalol or clofilium (-26 +/- 4 and -26 +/- 4 mV, respectively). Spontaneous EADs or EADs elicited by stimulation of Na-Ca exchange coincident with drug treatment were suppressed by increasing Ca02+ but were not suppressed by nifedipine (10(-7) M). CONCLUSION: EADs elicited by d,l-sotalol and clofilium in canine Purkinje tissue and epicardial myocytes are dependent upon Na Ca exchange rather than ICa-L "window current." PMID- 9209971 TI - Radiofrequency ablation-induced asystole during transaortic approach for a left anterolateral accessory pathway: a Bezold-Jarisch-like phenomenon. AB - We present a case of cardiac asystole induced by radiofrequency catheter ablation of a left anterolateral accessory pathway in a 28-year-old woman with Wolff Parkinson-White syndrome who was experiencing recurrent palpitation. Radiofrequency current applied on the ventricular aspect of the mitral annulus corresponding to the aforementioned site provoked profound slowing of the sinus rate preceded by disappearance of the preexcitation, and then asystole ensued. The proposed causal mechanism was a reflexogenically mediated hypotension bradycardia syndrome (Bezold-Jarisch-like phenomenon) through stimulation of either nearby vagal afferent pathways or sensory terminal receptors at the ablation site. PMID- 9209972 TI - Role of intracellular sodium overload in the genesis of cardiac arrhythmias. AB - A number of clinical cardiac disorders may be associated with a rise of the intracellular Na concentration (Na(i)) in heart muscle. A clear example is digitalis toxicity, in which excessive inhibition of the Na/K pump causes the Na(i) concentration to become raised above the normal level. Especially in digitalis toxicity, but also in many other situations, the rise of Na(i) may be an important (or contributory) cause of increased cardiac arrhythmias. In this review, we consider the mechanisms by which a raised Na(i) may cause cardiac arrhythmias. First, we describe the factors that regulate Na(i), and we demonstrate that the equilibrium level of Na(i) is determined by a balance between Na entry into the cell, and Na extrusion from the cell. A number of mechanisms are responsible for Na entry into the cell, whereas the Na/K pump appears to be the main mechanism for Na extrusion. We then consider the processes by which an increased level of Nai might contribute to cardiac arrhythmias. A rise of Na(i) is well known to result in an increase of intracellular Ca, via the important and influential Na/Ca exchange mechanism in the cell membrane of cardiac muscle cells. A rise of intracellular Ca modulates the activity of a number of sarcolemmal ion channels and affects release of intracellular Ca from the sarcoplasmic reticulum, all of which might be involved in causing arrhythmia. It is possible that the increase in contractile force that results from the rise of intracellular Ca may initiate or exacerbate arrhythmia, since this will increase wall stress and energy demands in the ventricle, and an increase in wall stress may be arrhythmogenic. In addition, the rise of Na(i) is anticipated to modulate directly a number of ion channels and to affect the regulation of intracellular pH, which also may be involved in causing arrhythmia. We also present experiments in this review, carried out on the working rat heart preparation, which suggest that a rise of Na(i) causes an increase of wall stress induced arrhythmia in this model. In addition, we have investigated the effect on wall stress-induced arrhythmia of maneuvers that might be anticipated to change intracellular Ca, and this has allowed identification of some of the factors involved in causing arrhythmia in the working rat heart. PMID- 9209973 TI - A preexcited left bundle branch block tachycardia: what is the tachycardia mechanism? PMID- 9209974 TI - Foldable intraocular lenses and vitreoretinal surgery. PMID- 9209975 TI - Accommodative intraocular lens: a challenge for future development. PMID- 9209976 TI - Treatment of suture track leak. PMID- 9209977 TI - Consultation section. Cataract surgical problem. PMID- 9209978 TI - Consultation section. Refractive surgical problem. PMID- 9209979 TI - Forceps-puncture continuous curvilinear capsulorhexis. AB - Forceps-puncture continuous curvilinear capsulorhexis uses a single instrument. Using less instrumentation during the procedure decreases the chance of inadvertent damage to ocular structures and reduces surgical time. Complications related to this technique are rare. PMID- 9209980 TI - Preventing expulsive hemorrhage using an anterior chamber maintainer to eliminate hypotony. AB - We describe a method in which constant infusion inflow through an anterior chamber maintainer (ACM) is used to maintain positive intraocular pressure (IOP) during cataract extraction through a self-sealing tunnel incision. A retrospective review of patient records showed that the difference in the incidence of intraoperative suprachoroidal hemorrhage was significantly greater in eyes in which IOP was not constant throughout surgery than in eyes in which IOP was maintained using the ACM system. PMID- 9209981 TI - Excimer laser photorefractive keratectomy for low hyperopia: safety and efficacy. AB - PURPOSE: To assess the safety and efficacy of photorefractive keratectomy (PRK) to correct low hyperopia. SETTING: University of Ottawa Eye Institute, Ottawa General Hospital, Ontario, Canada. METHODS: Twenty-five eyes with refractions of +1.00 to +4.00 diopters (D) and cylinder of 1.00 D or less were treated for hyperopia with the VISX Star excimer laser system using a refined ablation architecture. Thorough visual assessments were performed preoperatively (baseline) and 1, 3, and 6 months postoperatively. Complications were recorded and the level of patient satisfaction was noted. RESULTS: Mean spherical equivalent at 6 months was +0.27 D +/- 0.55 (SD), which was an 89% reduction over baseline. Eighty-four percent of patients gained two to seven lines of near uncorrected visual acuity (UCVA) and 1 patient (4%) lost more than one line. Eight percent achieved 20/25 or better UCVA. Approximately half realized their preoperative distance best corrected visual acuity (BCVA) by 1 month. By the end of the study, all patients had improved, achieved, or were within one line of their baseline distance BCVA. There were some slight reductions in lower contrast acuity at 6 months, although dim lighting conditions did not further reduce these acuities. Most patients had no clinically meaningful change in cylinder. The most common complications included early, transient corneal surface irregularities and visual symptoms and trace haze (grade < or = 0.5) in 14 of 23 patients at 6 months. All but 1 patient expressed a high degree of satisfaction. CONCLUSIONS: These results support the hypothesis that PRK shows great promise as a safe and effective treatment for low hyperopia. There were no significant complications and no decentered ablations. The slight regression occurred with or without the presence of trace haze. Overall, refractive stability was encouraging, although longer follow-up is needed. PMID- 9209982 TI - Topographical analysis of ablation centration after excimer laser photorefractive keratectomy and laser in situ keratomileusis for high myopia. AB - PURPOSE: To evaluate the ablation centration after photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) for high myopia and to assess the association between decentration and best corrected visual acuity (BCVA), glare, monocular diplopia, and halo phenomenon. SETTING: Mater Private Hospital, Dublin, Ireland. METHODS: Corneal topography was used to analyze centration in two groups of patients with myopia of more than 6.0 diopters: 18 had PRK and 18, LASIK. A standardized questionnaire assessed the preoperative and postoperative prevalence of glare, monocular diplopia, and halo phenomenon. RESULTS: "Significant" ablation decentration (0.5 mm) in the LASIK group (1.33 mm) was almost twice that in the PRK group (0.75 mm). Glare increased from 27% preoperatively to 42% in the PRK group; monocular diplopia increased in the LASIK group. Halo phenomenon decreased after both procedures. CONCLUSION: Laser in situ keratomileusis represents a step forward in the surgical correction of high myopia, but the accuracy of the corneal ablation location must be improved. Suction ring fixation of the globe or real time tracking systems may help improve centration. PMID- 9209983 TI - Topography after repair of full-thickness corneal laceration. AB - PURPOSE: To characterize corneal topography after repair of full-thickness corneal laceration. SETTING: Ophthalmic emergency room serving as a trauma referral center. METHODS: Twenty-two eyes with full-thickness corneal lacerations were prospectively studied after standardized surgical repair. Computerized videokeratography was done 2 and 14 weeks after surgery, with the latter measurement corresponding to 6 to 8 weeks after all sutures were removed. Fellow uninjured eyes served as the control group. RESULTS: Twenty eyes (91%) had a significant reduction in topographic distortion after suture removal. Mean corneal astigmatism, measured by simulated keratometry, was 10.70 diopters (D) +/ 5.90 D (SD) with sutures in place and 2.25 +/- 4.90 D after their removal (P < .005). Eighteen patients (82%) had 2.00 D or less of corneal astigmatism 6 to 8 weeks after all sutures were removed. The final distribution of topographic patterns was bow tie (50%), spherical/oval (36%), and irregular (14%). There was no significant correlation between laceration configuration (curvilinear, jagged, branched wound margins) and final topography. Lacerations that passed within 2.0 mm of the line of sight, however, were significantly more likely to have more than 2.00 D of final astigmatism. Mean central corneal power was 42.40 +/- 3.20 D in the injured eyes and 42.40 +/- 2.40 D in the uninjured fellow eyes. CONCLUSION: Although high astigmatism is frequently produced by corneal sutures used to repair full-thickness lacerations, the cornea has a substantial topographic memory that results in a marked normalization of contour after suture removal. PMID- 9209984 TI - Two-incision radial keratotomy for low myopia with astigmatism. AB - PURPOSE: To evaluate the effectiveness of two-incision radial keratotomy (RK) in correcting low-magnitude refractive myopic astigmatism. SETTING: Two clinical study sites, one in St. Louis, Missouri, USA, the other in Caracas, Venezuela. METHODS: Fifty-seven eyes of 43 patients with low-magnitude myopic astigmatism had two-incision RK at one of two clinical study sites. In the initial phase of this series, 10 eyes with amblyopia at the 20/30 level had surgery at one center. Refractive keratotomy was performed with the radial incision placed in the plus cylinder axis of refraction. This axis was verified as the meridian of greatest corneal curvature by standard keratometry and computer-assisted corneal topographic analysis. Two eyes received a second operation (enhancement). RESULTS: Mean follow-up was 11.1 months (range 6 to 12 months). Mean preoperative and postoperative myopic spherical equivalent measured -1.42 diopters (D) +/- 0.51 (SD) and -0.14 +/- 0.39 D, respectively; the mean reduction was 1.28 +/- 0.59 D (P = .0001). Mean preoperative and postoperative refractive astigmatism was 1.41 +/- 0.45 D and 0.48 +/- 0.33 D, respectively (P = .0001). Mean preoperative and postoperative keratometric astigmatism was 1.26 +/- 0.54 D and 0.31 +/- 0.35 D, respectively, a mean reduction of 0.95 D (P = .0001). The surgical meridian was flattened by an average of 2.06 D by keratometry and the orthogonal meridian, by an average of 1.10 D. Preoperative uncorrected visual acuity (UCVA) was 20/40 or better in five (9%) eyes (range counting fingers to 20/40). Postoperative UCVA acuity was 20/40 or better in all eyes (mean acuity 20/25). In the nonamblyopic subgroup mean postoperative UCVA was 20/24. CONCLUSIONS: A limited number of radial incisions placed in the topographically confirmed axis of greatest curvature are effective in the treatment of low magnitude myopic astigmatism. PMID- 9209985 TI - Corneal thickness variation during eight-incision radial keratotomy. AB - PURPOSE: To determine the degree of corneal thickness variability that may be encountered during routine radial keratotomy (RK) surgery. SETTING: Office refractive surgical suite. Laurel Eye Clinic, Brookville, Pennsylvania. METHODS: This study statistically analyzed variability in corneal thickness measured at the optical zone in 140 consecutive eight-incision RK cases. Pachymetry measurements were assessed at each of the eight circumferential optical zone locations and then evaluated in regard to intra-patient variation by location and inter-patient variation in location thickness patterns. Covariants such as central corneal thickness and differing optical zone size were also analyzed. RESULTS: Mean corneal thickness (adjusted for optical zone and central thickness) between the three superior locations and the three inferior locations varied by 10 microns. In more than 10% of cases, this adjusted difference was greater than 30 microns. Central corneal thickness and differing optical zone sizes had a significant effect on the results. CONCLUSION: Corneal thickness measured at the optical zone during eight-incision RK varied significantly by incision location, bringing into question the notion that one blade depth setting is adequate for all incisions. PMID- 9209986 TI - Histologic analysis of thermal effects of laser thermokeratoplasty and corneal ablation using Sirius-red polarization microscopy. AB - PURPOSE: To evaluate how well several histologic techniques differentiate degrees of thermally induced changes in corneal tissue after laser thermokeratoplasty (LTK) or corneal ablation. SETTING: Medical Laser Center Lubeck, Germany. METHODS: Corneas of freshly enucleated porcine eyes were treated with a continuous wave laser diode (1.86 microns) and a pulsed chromium-thulium-holmium: YAG laser (2.1 microns) to produce LTK lesions or ablated with a Q-switched and a free-running chromium-erbium: YSGG laser (2.70 microns), a free-running erbium: YAG laser (2.94 microns), and an argon-fluoride excimer laser (193 nm). The lesions were evaluated by light microscopy (LM) (hematoxylin and eosin, Azan, van Gieson's, and Masson-Goldner's trichrome stains), transmission electron microscopy (TEM), and polarization microscopy after Sirius-red staining. Sirius red, a strongly elongated, birefringent molecule binding parallel to collagen molecules, was used to enhance corneal birefringence. RESULTS: With routine LM, it was difficult to discriminate the degrees of thermal alterations in LTK lesions. Combined Sirius-red staining and polarization microscopy distinguished between a strongly coagulated central zone and the transition zone to normal tissue. Sirius-red uptake was increased in both zones, reflecting the availability of new binding sites. The central zone appeared darker under polarization than normal collagen because of a loss of birefringence. Intrinsic birefringence was greatly reduced; however, form birefringence partly remained as long as some collagen fibrils were intact. In the center of very strong lesions, where the collagen was hyalinized, birefringence was completely lost because of the complete disintegration of the fibrillar structure, which was visible under TEM. The transition zone toward normal cornea showed increased birefringence because the natural birefringence was largely preserved and enhanced by the increased Sirius-red uptake. Mechanical stretching between neighboring LTK lesions was manifested by increased birefringence. CONCLUSION: Sirius red offered an improved and simple histologic method for analyzing thermal collagen changes. It may contribute to a better understanding of the working mechanisms of LTK and improve analysis of thermal effects in corneal ablation. PMID- 9209987 TI - Practice styles and preferences of ASCRS members--1996 survey. AB - A survey of the practice styles and preferences of members of the American Society of Cataract and Refractive Surgery with a United States ZIP code was performed in September 1996. Approximately 26% (1440) of the 5520 questionnaires mailed were returned by the November cutoff date. Three main profile questions were used to cross-tabulate data: age of the ophthalmologist, geographic location, and volume of cataract surgery per month. Current data were compared with data in previous annual surveys. PMID- 9209988 TI - Silicone oil adhesion to intraocular lenses: an experimental study comparing various biomaterials. AB - PURPOSE: To perform an in vitro experimental study comparing the degree of adherence of silicone oil to various rigid and foldable intraocular lens (IOL) designs and to the human lens capsule. SETTING: Center for Research on Ocular Therapeutics and Biodevices, Department of Ophthalmology, Storm Eye Institute, Medical University of South Carolina, Charleston, South Carolina, USA. METHODS: Seven IOL styles comprising various biomaterials were studied: fluorine-treated (Fluorlens), heparin-surface-modified (HSM), hydrogel, Memory-Lens, Poly(methyl methacrylate) (PMMA), soft acrylic, and silicone lenses; the human crystalline lens was also studied. Each lens was immersed in silicone oil for 12 hours, than photographed, studied by scanning electron microscopy (except the crystalline lens), and subjected to computer-generated image analysis to determine the silicone oil coverage. RESULTS: Silicone oil coverage of dry silicone lenses was 100% and of lenses immersed in normal saline, 82.5%. The least coverage was on the heparin-surface-modified lens (mean score 9.4%). Coverage of the other four lenses ranged from approximately 15.1% to 33.7%. Mean coverage of the human lens capsule was 10.9%. CONCLUSION: Although a silicone IOL shows maximal adherence to silicone oil, other lens biomaterials are not immune to this complication. Silicone oil coverage was related to the dispersive energy component of the surface charge of the IOL biomaterial. Low dispersive energy materials had less silicone oil coverage, while those with higher dispersive energy had more oil coverage. PMID- 9209989 TI - Unpreserved lidocaine to control discomfort during cataract surgery using topical anesthesia. AB - PURPOSE: To determine whether intraoperative unpreserved lidocaine further decrease discomfort or pain during sutureless small incision cataract surgery and intraocular lens (IOL) implantation under topical anesthesia. SETTING: Outpatient ambulatory surgical center. METHODS: In this prospective controlled study, comparable eligible patients were randomized to receive 0.1 cc unpreserved lidocaine 1% or 0.1 cc balanced salt solution (BSS) (control group) in double masked fashion. Study drugs were injected intracamerally 1 minute before phacoemulsification. A predefined uniform pain/discomfort scale was used for assessment during phacoemulsification and IOL insertion. A secondary study using a 0.5 cc dose was also performed. RESULTS: Twenty-six percent in the control group and 9% in the lidocaine group had discomfort pain scores of 2 or more; 10% in the BSS group felt increased pressure or pain during phacoemulsification. In the lidocaine group, discomfort was felt mainly during IOL insertion, possibly as a result of wound manipulation. During phacoemulsification, no patient in the lidocaine group reported pain; 2% felt increased pressure during phacoemulsification. A dose increase to 0.5 cc reduced any intraocular sensation to 3% in the lidocaine group. No patient in either group had significant cell loss or adverse events. CONCLUSION: Intraoperative lidocaine is safe and effective in controlling intraoperative discomfort. PMID- 9209990 TI - Anterior chamber irrigation with unpreserved lidocaine 1% for anesthesia during cataract surgery. AB - PURPOSE: To assess the effectiveness of using anterior chamber irrigation of unpreserved lidocaine 1% as anesthesia during cataract surgery. SETTING: Private group ophthalmology practice. METHODS: This study prospectively evaluated 1000 of 1012 consecutive eyes having temporal corneal incision cataract surgery to determine whether anterior chamber lidocaine provides adequate anesthesia for cataract surgery. Twelve eyes were excluded because the patients had preoperative sedation. Surgery on the remaining 1000 eyes was performed by one surgeon without patients receiving preoperative or intraoperative sedation or other medications other than the local anesthetic and dilating agents. Each received one drop of topical proparacaine before entering the operating room. After an initial corneal stab incision was made, 0.25 to 0.50 cc of unpreserved lidocaine 1% was irrigated into the anterior chamber. RESULTS: One patient was so uncomfortable from the microscope that he required supplemental retrobulbar anesthesia. A second patient was extremely uncomfortable during the case but did not require supplemental anesthesia. Two patients received additional dosages of anesthetic because of discomfort late in the operation. The remaining 996 patients were comfortable and pain-free during the operation with a single dose of the anesthetic. CONCLUSION: Anterior chamber irrigation with unpreserved lidocaine 1% was an effective method for anesthetizing an eye for temporal corneal incision cataract surgery. PMID- 9209991 TI - Computer-assisted videokeratography to measure changes in astigmatism induced by sutureless cataract surgery. AB - PURPOSE: To compare the visual outcome, change in surgically induced astigmatism, and corneal thickness after sutureless cataract surgery through either a scleral tunnel or clear corneal incision. SETTING: Department of Ophthalmology, Kangnam St. Mary's Hospital, Catholic University Medical College, Seoul, Korea. METHODS: This retrospective study evaluated 79 eyes of 64 patients who had cataract surgery using a 6.0 mm scleral tunnel incision (Group 1, n = 20 eyes), 3.1 mm clear corneal incision at the 10 o'clock position (Group 2, n = 35 eyes), or 3.1 mm clear corneal incision at the 10 o'clock position (Group 3, n = 24 eyes). Changes in surgically induced astigmatism were analyzed using computer-assisted videokeratography (CVK) 1 day, 8 weeks, and 6 months postoperatively. RESULTS: Eleven eyes (55.0%) in Group 1, 15 (42.8%) in Group 2, and 11 (48.5%) in Group 3 had an uncorrected visual acuity of 20/40 or better 1 day postoperatively. The CVK measurements showed significantly more flattening at radial distances of 0.75, 1.50, and 2.50 mm along the 90 degree semimeridian in Group 2 than in Group 1 (P < .05) 1 day after surgery; in Group 3, flattening along the incisional semimeridian fell between the amounts in Groups 1 and 2. At 8 weeks, the amount of flattening decreased in all groups, and the differences between groups were not statistically significantly different (P > .05). In the CVK pattern, corneal flattening along the incisional meridian was narrower and longer in Groups 2 and 3 than in Group 1. CONCLUSION: Localized flattening along the incisional meridian was prominent temporally after clear corneal incision surgery. However, there was no difference in early visual rehabilitation between the clear corneal and scleral tunnel incision groups. PMID- 9209992 TI - Cataract formation after penetrating keratoplasty. AB - PURPOSE: To assess the incidence and risk factors for developing cataract after penetrating keratoplasty (PKP). SETTING: L.V. Prasad Eye Institute, Hyderabad, India. METHODS: This retrospective analysis of 251 phakic patients who had PKP between 1987 and 1994 assessed the incidence of and risk factors for cataract formation. Patients were assigned to one of two groups: Group 1 (n = 96), patients with keratoconus and corneal dystrophy; Group 2 (n = 88), patients with corneal scar and adherent leucoma. Preoperative, intraoperative, and postoperative lens details were noted. Data on intraoperative iris procedures (excess manipulation, pupilloplasty, synechiolysis, peripheral iridectomy) and postoperative topical steroid usage were assessed. RESULTS: Sixty-seven patients were excluded because of incomplete lens status data. Of the remaining 184 patients, 45 (24.45%) developed cataract. Most cataracts (n = 31) developed within the first year of surgery. The incidence of cataract was significantly higher in Group 2 (n = 29) than in Group 1 (n = 16) (P = .0102). There was no significant between-group difference in mean steroid dose (P = .7064); however, the mean dose was significantly higher in eyes with cataracts (563 +/- 234 units) than in those without (479 +/- 127 units) (P = .0352). In Group 2, 9 of 20 patients who had synechiolysis, 1 of 3 who had pupilloplasty, and 2 of 5 who had peripheral iridectomy developed cataract. In Group 1, no patient had iris-related procedures. CONCLUSION: Excessive steroid use and intraoperative iris manipulations are major risk factors for cataract formation after PKP. PMID- 9209993 TI - Mitomycin-C supplemented trabeculectomy, phacoemulsification, and foldable lens implantation. AB - PURPOSE: To evaluate the outcome of combined mitomycin-C filtering surgery, phacoemulsification, and foldable intraocular lens (IOL) implantation. SETTING: Wills Eye Hospital, Thomas Jefferson University, Philadelphia, Pennsylvania, USA. METHODS: This retrospective study evaluated 182 eyes of 174 patients who had combined mitomycin-C trabeculectomy, phacoemulsification, and insertion of a foldable IOL through a 3.5 mm incision. Success of the combined procedure was defined as intraocular pressure (IOP) below 21 mm Hg, with or without medications, and no serious complication. Success rates were calculated using the Kaplan-Meier actuarial method. RESULTS: Mean follow-up was 16.7 months +/- 5.4 (SD). The probability of success at 6, 12, 18, and 24 months was 98.3, 95.6, 90.6, and 88.0%, respectively. When compared with preoperatively, visual acuity improved one or more lines in 148 eyes (81.3%) and worsened one or more lines in 15 (8.2%); 111 eyes (61.0%) achieved visual acuity of 20/40 or better. The most frequent complication was posterior capsule opacification requiring capsulotomy, which occurred in 22 cases (12.0%). CONCLUSION: The 1 year and 2 year IOP control rate of combined mitomycin-C filtering procedures and phacoemulsification in glaucoma patients was high. PMID- 9209994 TI - Risk factors, complications, and results in extracapsular cataract extraction. AB - PURPOSE: To examine a large series of extracapsular cataract extractions (ECCEs) to determine the risk factors for posterior capsule rupture with vitreous loss (PCR + VL) and the complications and results of this cataract surgery technique. SETTING: North Riding infirmary, Middlesbrough, United Kingdom. METHODS: In this retrospective study, the records of 1552 patients who had ECCE performed by two surgeons were examined. Follow-up was 4 months to 4 years. The main outcome measures were the incidence of risk factors, PCR + VL, major and minor postoperative complications, and the visual outcome. RESULTS: Three hundred twenty-two cases were considered high risk for PCR + VL. In this group, the PCR + VL rate was 5.3%; it was 3.7% in the low-risk group and 4.0% overall. Visual acuity of 6/9 or better was achieved by 76% of all eyes. Postoperative complications were 3 cases of endophthalmitis, 4 of retinal detachment, and 1 of pseudophakic bullous keratopathy. In 323 eyes, loose or broken sutures were removed and in 175, a neodymium: YAG capsulotomy was performed because of posterior capsule opacification. CONCLUSIONS: The incidence of perioperative and postoperative complications was comparable with those reported in other series. Factors that increased the risk of PCR + VL included pseudoexfoliation, diabetes mellitus, and a traumatic etiology. Previous glaucoma surgery and axial myopia of greater than 26.0 mm did not increase the PCR + VL risk. Loose or broken corneal sutures was a common finding that could be reduced substantially by planned suture removal. PMID- 9209995 TI - Postoperative cataract surgery satisfaction in a rural Kenyan clinic. AB - PURPOSE: To assess prospectively the factors influencing patient satisfaction following intracapsular cataract extraction (ICCE) surgery in a rural eye unit in Kenya. SETTING: Nakuru Eye Unit, Rift Valley Provincial General Hospital, Nakuru, Kenya, and Johns Hopkins School of Public Health, Department of International Health, Baltimore, Maryland, USA. METHODS: Starting in November 1992, 232 consecutive blind or visually impaired rural patients, over age 40, with simple senile cataract were offered free standard ICCE. Only 70% agreed to surgery. An interviewer-administered questionnaire and a brief interview were performed postoperatively on day 2, completing an extensive preoperative analysis that was part of the Kenya Rural Cataract Project. A satisfaction level indicator composed from the most important factors, applying a logistic regression model, is suggested as a predictive index for a patient to become a motivator in his or her community. RESULTS: Most patients were happy with their decision to have cataract surgery, even though 92% of the operations were done by clinical officers. Patients were overwhelmingly willing to have their fellow eye operated on or to recommend the operation to another "blind" friend (83.4%). The proposed model correctly classified 87.1% of operated patients, with high sensitivity (88.2%) and specificity (81.5%). CONCLUSIONS: Cultural differences are paramount in determining health behavior priorities and satisfaction. The post ICCE satisfaction in developing countries must be better evaluated to achieve higher self-referral of cataract-blind patients for surgery in Africa. Hospital conditions, although appreciated, did not play a major role in patients' satisfaction. The immediate surgical outcome was the key factor. PMID- 9209996 TI - Day-case cataract surgery in rural Spain. AB - OBJECTIVE: To ascertain how many patients in a rural area of Spain would qualify for and choose to have day-case cataract surgery. SETTING: Departmento de Oftalmologia, Hospital Comarcal, Hellin, Albacete, Spain. METHODS: All patients intending to have cataract surgery in 1993 responded to a five-question survey. Only patients answering yes to all five questions were considered candidates for day-case surgery. RESULTS: Thirty-three of 374 patients (9.0%) answered yes to all five questions. Only 7 of the 33 (1.9%) subsequently had day-case surgery. CONCLUSION: Establishing a day-case cataract surgery, unit in rural areas requires consideration of factors not present in urban areas and may require more time for patient acceptance. PMID- 9209997 TI - Cataract surgery and lens implantation in eyes with exfoliation syndrome. AB - OBJECTIVE: To evaluate the surgical technique, intraoperative complications, and outcomes of extracapsular cataract extraction (ECCE) and intraocular lens (IOL) implantation in patients with exfoliation syndrome. SETTING: Department of Ophthalmology, Hippokration Thessaloniki General Hospital, Thessaloniki, Greece. METHODS: This prospective study comprised 84 patients with exfoliation syndrome who had ECCE and posterior chamber IOL implantation between December 1992 and November 1993. Main outcome measures were poor mydriasis, zonular rupture, vitreous loss, and best corrected visual acuity. Mean follow-up was 9.32 months (range 4 to 16 months). RESULTS: Pupil diameter was smaller than 5.0 mm in 61.90% of the eyes. Zonular rupture, capsular tear, and vitreous loss occurred in 13.09, 10.71, and 7.14%, respectively. Best corrected visual acuity ranged from 7/10 to 10/10 in 72 eyes (85.71%). CONCLUSION: Patients with exfoliation who had ECCE and posterior chamber IOL implantation had an increased risk of intraoperative complications. In these patients, surgery must be done by experienced surgeons able to handle all possible complications. PMID- 9209998 TI - Influence of intraocular lens haptic material on bacterial isolates from anterior chamber aspirate. AB - PURPOSE: To determine whether intraocular lens (IOL) type affects the bacterial count in anterior chamber aspirates obtained immediately after cataract surgery. SETTING: L.V. Prasad Eye Institute, Hyderabad, India. METHODS: This in vivo study evaluated two groups of eyes that had uneventful cataract extraction and implantation of one of two types of IOLs: all poly(methyl methacrylate) (PMMA) (n = 73) or polypropylene haptic (n = 83). Anterior chamber fluid aspirates (0.1 mL) were obtained with a 30 gauge cannula at the end of surgery and inoculated onto chocolate agar and in thioglycolate broth. Microbiology evaluation was performed using standard methods. RESULTS: Seven eyes (9.5%) with all-PMMA IOLs and 21 (25.3%) with polypropylene haptic IOLs were positive for bacterial isolates (P = .0322; chi-square test). Mean colony count (+/-SD) was 11.43 +/- 3.78 and 13.16 +/- 4.78 colony-forming units/ milliliter in the PMMA and polypropylene haptic IOL groups, respectively. No eye developed endophthalmitis. CONCLUSION: Polypropylene haptic IOLs were associated with a significant increase in bacteria in the anterior chamber. PMID- 9209999 TI - Refractory intraocular pressure increase after photorefractive keratectomy. AB - A patient with developmental angle anomaly developed a corticosteroid-induced refractory increase in intraocular pressure (IOP) after photorefractive keratectomy (PRK). Trabeculectomy was required to reduce the pressure. Although rare, corticosteroid-induced refractory IOP increase is a serious complication of PRK and may necessitate trabeculectomy. More frequent monitoring of IOP in post PRK patients and a re-evaluation of postoperative treatment are indicated. PMID- 9210000 TI - Transscleral ciliary sulcus fixation of a posterior chamber lens in an eye with congenital aniridia. AB - A 30-year-old man with bilateral congenital aniridia presented with a subluxated cataract in the right eye. He wore a soft contact lens for aphakic correction in the fellow eye after lensectomy for a subluxated cataract. We performed pars plana lensectomy and vitrectomy with implantation of a transscleral ciliary sulcus fixated posterior chamber intraocular lens in the right eye. Good postoperative visual acuity was obtained. PMID- 9210001 TI - Amoxycillin clavulanate: an assessment after 15 years of clinical application. PMID- 9210002 TI - Potential benefit of acyclovir for chickenpox acquired from household contacts. The Italian Acyclovir-Chickenpox Study Group. AB - As a part of a trial of acyclovir treatment of chickenpox in otherwise healthy children, the assessment of disease features and evolution showed a significantly more severe clinical picture at onset in the 215 patients with intrafamiliar exposure to varicella, compared with the remaining 486 with community-acquired infection, although the disease course proved similar by the third day of antiviral therapy. Children with household exposure to varicella are likely to suffer from a more severe disease, and might especially benefit from acyclovir treatment. PMID- 9210003 TI - Effect of norfloxacin, trimethoprim-sulfamethoxazole and nitrofurantoin on fecal flora of women with recurrent urinary tract infections. AB - The effects of antibiotics used for prophylaxis in women with recurrent urinary tract infections (UTIs) on the aerobic intestinal flora were investigated. Twenty one patients with recurrent UTIs were randomly divided into three groups. The patients of each group received monotherapy with oral norfloxacin, trimethoprim sulfamethoxazole or nitrofurantoin for one month. Urine and stool quantitative aerobic cultures were performed before prophylaxis, 2 and 4 weeks after the initiation of therapy, and 2 weeks after antibiotics were discontinued. The gram negative aerobic flora was strongly suppressed during the administration of norfloxacin and trimethoprim-sulfamethoxazole, while Enterococcus spp. were not affected. Resistant strains of Escherichia coli were detected in two patients, one in the norfloxacin and one in the trimethoprim-sulfamethoxazole group. The aerobic intestinal flora was not affected by nitrofurantoin. These findings help in the selection of the most appropriate antimicrobial agent for prophylaxis in recurrent UTIs, so as to reduce the possibility of emergence of resistant bacterial strains. PMID- 9210004 TI - Comparison of colloidal bismuth subcitrate and metronidazole, both in combination with an H2-antagonist as therapy for Helicobacter pylori. AB - In this random study, the efficacy of either colloidal bismuth subcitrate (CBS) or metronidazole in combination with an H2-antagonist in the treatment of various gastric pathologies was evaluated, along with the trends in antibody levels. Among the 40 Helicobacter pylori-positive patients with various gastroduodenal pathologies who underwent chemotherapy, 27 were treated with CBS and 13 with metronidazole. H. pylori was eradicated in 48.14% of the patients treated with CBS and 53.8% of those treated with metronidazole. After therapy, no statistically significant or slight decrease in the serum levels of antibodies was found. PMID- 9210005 TI - Cefepime versus ceftazidime in the treatment of lower respiratory tract infections. AB - The objective of the study was to compare the safety and efficacy of cefepime and ceftazidime in the treatment of community acquired lower respiratory tract infections of moderate intensity. Eighty-six patients were randomized at a 2:1 ratio to receive respectively cefepime 1 g b.i.d. or ceftazidime 1 g t.i.d. The drugs were well tolerated and the occurrence of adverse events in each group was comparable. The rates of satisfactory clinical response were 96% (49/51) for cefepime and 89% (24/27) for ceftazidime. A total of 73 pathogens were isolated and pathogen eradication rates were 98% and 96% respectively for the cefepime and ceftazidime treatment groups. In conclusion, the data confirmed that cefepime could be a good alternative to ceftazidime. PMID- 9210006 TI - In vitro effects of lithium chloride on TNF alpha and IL-6 production by monocytes from breast cancer patients. AB - It is well known that lithium chloride (LiCl) is able to trigger human monocytes to release tumor necrosis factor alpha (TNF alpha). In this study we have evaluated the in vitro effect of LiCl on TNF alpha and interleukin-6 (IL-6) release by monocytes from patients affected by non-metastatic (BCa/M0) and metastatic breast cancer (BCa/M1), preincubated with autologous serum (sPt). Our data demonstrate that monocytes from cancer patients (BCa) treated with LiCl released lower amounts of TNF alpha compared to those from healthy donors (HD). Preincubation in autologous serum (sPt) impaired TNF alpha production by monocytes from BCa with LiCl. On the contrary, our data indicate that IL-6 production by monocytes treated was not impaired. Moreover, the results obtained from the same cells, preincubated in sPt and treated with LiCl, indicate that serum factors may synergize with LiCl treatment in releasing IL-6. PMID- 9210008 TI - Bacteremia in cancer patients with solid tumors undergoing chemotherapy versus surgery: risk factors, etiology and outcome in 276 patients. AB - Etiology, risk factors, outcome and complications of bacteremia in 276 patients with solid tumors were analyzed. A group of 78 patients with solid tumors and surgical therapy only was compared with 172 patients with solid tumors who were treated with chemotherapy only. The most frequently observed risk factors of bacteremia in patients after surgery was urinary catheter insertion, wound as source of bacteremia, age > 60, staphylococci, enterococci and Enterobacteriaceae as etiologic agents. In comparison, viridans streptococci and Pseudomonas aeruginosa as etiologic agents as well as vascular catheters were significantly more frequently found in those treated with chemotherapy only. Patients with bacteremia after surgery only had a lower incidence of septic shock (6.4 vs. 16.9%, P < 0.03) and also lower mortality (5.6 vs. 14.9%, P < 0.04) attributable to shock than patients being treated for solid tumors with chemotherapy only. PMID- 9210007 TI - Ceftriaxone as a single agent in empirical therapy of unexplained fever in granulocytopenic children with solid tumors. AB - The optimal management of fever in granulocytopenic cancer patients remains controversial. Antibiotic monotherapy is increasingly an option for the initial empiric treatment of febrile granulocytopenic patients with solid tumors. Available data show that response to empiric therapy is often more related to disease classification (solid tumors vs. acute leukemia) than to the regimen used. In this study we based empiric monotherapy on the underlying disease (solid tumors) in treating 33 episodes of fever in 26 granulocytopenic children with cancer. We investigated the potential effectiveness of single daily doses of ceftriaxone administered empirically in febrile granulocytopenic children with solid tumors. Fever was treated successfully with ceftriaxone monotherapy in 91% (30/33) of febrile episodes. None of the patients died as a result of primary infection. These results suggest that empirical monotherapy with once-daily ceftriaxone is safe and effective. In addition, when compared with other extended spectrum cephalosporins such as ceftazidime, once-daily administration of ceftriaxone reduces cost and patient inconvenience, allowing convenient parenteral therapy even on an outpatient basis. PMID- 9210010 TI - Lesbian coming out as a multidimensional process. AB - Coming out for lesbians is discussed as a multidimensional process. Four dimensions are presented and explored. First is sexual identity formation which encompasses development of lesbian sexuality and awareness of being a lesbian. Second is disclosure of sexual orientation to others. Third is sexual expression and behavior. Fourth is lesbian consciousness which refers to how lesbians see themselves in relation to the social environment, including lesbian and gay communities. These dimensions are examined in relation to existing theories and research about lesbian coming out processes. Further, there is a discussion of coming out and demographic differences among lesbians. The possible impact of age, race, ethnicity, religion, geographic location, income, employment, and education on lesbian coming out are considered. PMID- 9210011 TI - The relationship of homophobia to intimacy in heterosexual men. AB - Interpersonal intimacy is more difficult to achieve for American men than women. Research has shown that men disclose less, have fewer close friendships, and are viewed by their wives as low in intimacy. Among the barriers to intimacy among men, Tognoli (1980) suggested that homophobia is the most powerful. The present study tested this idea by asking men to disclose a personal secret to either a male confidant, a female confidante, or by writing the secret. It was expected that the presence of a male confidant would stimulate homophobic feelings and inhibit disclosure, while the other two conditions would result in more personal disclosure. A correlation between intimacy of disclosure and level of homophobia was also expected. Subjects were 75 heterosexual men, 31 to 50 years of age, randomly assigned to one of the three conditions. They also completed the Miller Social Intimacy Scale, the Jourard Self-Disclosure Questionnaire, the Index of Homophobia, and the Marlowe-Crowne Social Desirability Scale. Results showed that homophobia was inversely related to level of social intimacy. There was some indication that homophobia does relate to less intimate disclosure to other males. Thus this study found some support for the idea that homophobia is an obstacle to intimacy for men. PMID- 9210009 TI - Aztreonam/clavulanic acid in the treatment of serious infections caused by Stenotrophomonas maltophilia in neutropenic patients: case reports. AB - Two seriously neutropenic patients (a 23-year-old man with a promyelocytic acute myeloid leukemia [AML-M3] and a 77-year old male with an immature acute myeloid leukemia [AML-M1] diagnosis) with severe infections caused by Stenotrophomonas maltophilia were treated with aztreonam/clavulanic acid (2:1) combination. In the first patient the infection was caused by a multiresistant strain and in the second, by a strain with poor response to trimethoprim-sulfamethoxazole and other antimicrobial agents. After treatment with aztreonam/clavulanic acid both patients evolved favorably. PMID- 9210012 TI - The impact of social support on gay male couples. AB - The impact of social support on the relationships of gay male couples is examined. Social support for couples is defined as the availability of people outside the couple perceived as willing to offer resources in a manner that provides assistance and validates the couple's relationship. The sample consisted of 156 cohabiting gay male couples. It was found that social support for the couple is positively correlated with relationship quality. There are promising but not definitive indications that social support for the couple better predicts relationship quality than does social support for the individual. The categories of social support providers having a unique impact on relationship quality were family members and friends. PMID- 9210013 TI - Changes in attitudes toward homosexuality in college students: implementation of a gay men and lesbian peer panel. AB - The purpose of this study was to evaluate the implementation of a gay men and lesbian peer panel as an educational strategy in changing homophobic attitudes among 190 college students. The students were administered a modified version of the Attitudes Toward Homosexuality Scale before and after the intervention. Results indicated a significant difference between pre- and posttest scores. Men had more negative attitudes toward homosexuals than did women both before and after intervention. Suggestions for counselors, student development professionals, and other human service professionals pertaining to the delivery and format of the intervention are discussed. PMID- 9210014 TI - Assessing homosexual stress. AB - The present study was designed to replicate and extend an existing technique for assessing homosexual stress using Personal Construct Theory. In addition to using difference scores between present and homosexual self-ratings on subjects' personal construct rating dimensions to determine homosexual stress, difference scores between ideal and homosexual selves were also studied to determine if this method would represent a more accurate assessment of homosexual stress. Levels of homosexual stress were obtained for each subject from each technique and were correlated with attitudes toward homosexuality. The influence of construct meaningfulness and level of homosexual stress on attitudes toward homosexuality was also explored to determine if higher stress on more meaningful constructs yields more negative attitudes. The results of this study revealed that each stress technique was significantly and equivalently positively correlated with negative attitudes toward homosexuality. Homosexual stress was found to be independent of the meaningfulness measures used but positively correlated with authoritarianism. PMID- 9210015 TI - Sydney Anglicans and homosexuality. AB - The city of Sydney in Australia has one of the largest gay and lesbian communities in the English-speaking world, while the Anglican Church in Sydney is proud of its reputation as a strong-hold and guardian of conservative evangelicalism. Since the early 1970s the Anglican diocese, in its official statements and pastoral policies, has been strongly opposed to homosexuality and the organized gay movement. In 1973 a report on homosexuality by its Ethics and Social Questions Committee was unusual at the time because it recommended the continuance of legal sanctions against male homosexual behavior. There have been many confrontations between Anglican institutions and Sydney's increasingly confident gay community. This paper examines these tensions, the reasons for the stance of the diocese, and the responses of gay Christian groups such as Cross+Section and AngGays. PMID- 9210016 TI - Using distance matrices to choose between competing theories and an application to the origin of modern humans. AB - This paper examines competing theories for cases in which both the data and the hypotheses can be represented as distance matrices. A test due to Dow & Cheverud has been used for such comparisons in anthropology, but when data are spatially, temporally, or phylogenetically autocorrelated, this test may be far too liberal. We examine a classification procedure based on ratios of probabilities obtained from Mantel tests of the competing hypotheses and find that design matrices describing only lag-one connections and those eliminating common connections of competing hypotheses are the most informative. We apply this method to simulated gene-frequency data in a 7 x 7 chessboard representing a stepping-stone model and discriminate between alternative theories with a 7% misclassification rate. We also apply these techniques to the current controversy concerning the origin of anatomically modern humans by testing design matrices representing regional continuity and single African origins. The outcome for lag-one matrices and those showing only unique lag-one differences indicate that the single African origin of anatomically modern humans fits the distance matrix based on 165 characters of 83 fossil crania better than the competing theory. However, we also tested a design matrix describing single origin out of southwest Asia. This design matrix was clearly most similar to the data in all tested cases. These results make the regional-continuity theory a less likely explanation for the observed cranial differences than the two single-origin theories. Of these, single southwest Asian origins seems the more likely interpretation of the data. PMID- 9210017 TI - Body mass in comparative primatology. AB - Data are presented on adult body mass for 230 of 249 primate species, based on a review of the literature and previously unpublished data. The issues involved in collecting data on adult body mass are discussed, including the definition of adults, the effects of habitat and pregnancy, the strategy for pooling data on single species from multiple studies, and use of an appropriate number of significant figures. An analysis of variability in body mass indicates that the coefficient of variation for body mass increases with increasing species mean mass. Evaluation of several previous body mass reviews reveals a number of shortcomings with data that have been used often in comparative studies. PMID- 9210018 TI - Chronological changes in stone tool assemblages from Krapina (Croatia). AB - This study presents the results of the first recent analysis of stone tool assemblages from Krapina (Croatia). All assemblages are Pleistocene in age and many are associated with human remains, the Krapina Neandertals. The assemblages are described typologically and technologically, and subtle chronological changes in raw material selection and technology of tool blank production are observed. These changes involve increasingly sophisticated and selective use of lithic materials. Changing artefact assemblages are considered in light of variability in the hominids from Krapina, and are interpreted as reflecting behavioral change among Neandertals rather than between Neandertal and modern human populations. PMID- 9210019 TI - Re-analysis of the hominid radii from Cave of Hearths and Klasies River Mouth, South Africa. AB - Two of the few postcranial fragments from the late Early Stone Age and/or the Middle Stone Age of southern Africa are the proximal radii from the Cave of Hearths and Klasies River Mouth. The Cave of Hearths fossil is metrically indistinguishable from both archaic (e.g., Neandertals) and recent humans, and presents a mosaic of primitive and modern features. The primitive include a relatively slender neck and thick cortical bone (the latter of which distinguishes recent humans from archaic, Early Modern, and Upper Paleolithic hominids); the modern includes an anteromedially (rather than medially) facing radial tuberosity. Its extreme collo-diaphyseal angle is unusual, although it can be matched by modern homologues. The neck-shaft angle of some Neandetral and Early Modern radii also appears to match that of the Cave of Hearths specimen. The Klasies River Mouth radius also has thick cortical bone of the neck. It is morphologically indistinguishable from Early Modern and Neandertal homologues. These, and other fossils, suggest a mosaic pattern of evolution in the postcranial skeleton of the late Early Stone Age and/or Middle Stone Age inhabitants of sub-Saharan Africa. PMID- 9210020 TI - Social pressures have selected for an extended juvenile period in primates. AB - Primates are highly social animals. As such, they utilize a large repertoire of social skills to manage their complex and dynamic social environments. In order to acquire complex social skills, primates require an extended learning period. Here 1 perform a comparative analysis using independent contrasts to show that social pressures have favored an extension in the proportion of time primates spend as juveniles. PMID- 9210021 TI - Pertussis toxin induces lymphocytosis in rhesus macaques. AB - Lymph nodes and other solid tissues of the immune system are the principal sites for antigen presentation and lymphocyte activation. Lymphocytes in peripheral blood recognize the high endothelial venules within lymphoid tissues and cross from blood to tissue by the process of extravasation. Pertussis toxin is known to block extravasation and cause lymphocytosis in murine models but has not been studied extensively in nonhuman primates. We used intravenous injection of soluble pertussis toxin to induce a transient lymphocytosis in rhesus monkeys. The increase in total white blood cells was proportionally greater for lymphocytes than for polymorphonuclear cells and the CD4+ lymphocyte subpopulation increased more than the CD8+ cell population. The presence of immature polymorphonuclear cells suggested some activation of bone marrow. Clinical chemistry studies revealed an effect of pertussis toxin on liver function. Pertussis toxin is a powerful immunomodulatory agent that can disrupt and reorganize solid lymphoid tissues. PMID- 9210022 TI - The glomerular filtration rate, effective renal plasma flow, and renal blood flow in the adult male chacma baboon (Papio ursinus). AB - The radionuclide determination of glomerular filtration rate (GFR) and effective renal plasma flow (ERPF) has been validated in man, but not in the primate. GFR, ERPF, and renal blood flow (RBF) were measured in a group of 12 adult male chacma baboons using radiopharmaceuticals. GFR was determined using 99mtechnetium labelled diethylenetriamine-pentacetic acid. ERPF was measured with 131iodine labelled hippuran. RBF, body surface area, and kidney weights were calculated using standard formulae. GFR was 49 +/- 11 ml/min and ERPF was 237.9 +/- 54.2 ml/min. Calculated RBF was 430.7 +/- 111.9 ml/min and 507.4 +/- 138.4 ml/min/100 g of renal tissue. The results are in agreement with those obtained using more laborious nonradioisotopic techniques such as para-aminohippurate (PAH) and creatinine clearance and could serve as baseline normal values in the adult male chacma baboon. PMID- 9210024 TI - Abnormal morphology of vervet monkey sperm. AB - Ejaculates of 14 colony-bred and 14 wild-caught vervet monkeys were examined for morphologically abnormal sperm. Sperm head abnormalities were rare in both groups, occurring at rates of 0.01-0.29%. Tail abnormalities predominated, particularly bent midpieces and coiled and folded tails, which all occurred at rates of 3.79-17.18%. Except for the nipple acrosome, there was no difference in the rate at which sperm abnormalities were found in both groups. Three abnormalities were found only in colony-bred and three only in wild-caught individuals. Some common abnormalities, all affecting the tail, were highly variable in consecutive ejaculates from the same individuals. PMID- 9210023 TI - Hormonal response to restraint in rhesus monkeys. AB - The purpose of this study was to characterize the hormonal responses to a restraining system in four adult male rhesus monkeys (Macaca mulatta) in preparation for a spaceflight project. After the monkeys were accustomed to food and water (Phase I), blood-volume-regulating hormones were measured during three phases: 10 days in a metabolic cage (Phase II), 16 days sitting in a restrained position in a specially designed metabolism chair (Phase III) and 10 days in metabolic cage (Phase IV). An increase of active renin (30%) and vasopressin (25%) was observed at the end of Phase III. A decrease of atrial natriuretic peptide (ANP), urodilatin, and sodium excretion occurred during the first days of Phase III. Catecholamines were unchanged. A dramatic increase (tenfold) in urinary excretion of growth hormone occurred during all of Phase III and at the beginning of Phase IV. These findings are similar to those found in man during isolation inactivity and during confinement stress. PMID- 9210026 TI - Inflammatory arthritis in Pongo. AB - As the arboreal ape, Pongo, has an unusual ground ambulation adaptation, it was of interest to assess the impact of Pongo gait on patterns of arthritis. While osteoarthritis was not identified in Pongo, 11% of individual skeletons were afflicted with an inflammatory, erosive arthritis. The presence of sacroiliac involvement and the nature and distribution of erosive lesions allowed definitive diagnosis of spondyloarthropathy. Character, distribution, and radiologic appearance revealed a picture distinguishable from spondyloarthropathy in other primates. PMID- 9210025 TI - Characterization of dermatologic changes in geriatric rhesus macaques. AB - Nonhuman primates are frequently used for aging studies. We observed a high prevalence of skin disease among a group of geriatric rhesus monkeys (mean age = 25 years; n = 9) used in aging behavioral studies. Gross and histopathologic changes in the skin of these geriatric rhesus were compared with skin from control adult monkeys (mean age = 10; n = 4) and sun-exposed monkeys (mean age = 11; n = 4) to characterize age-related skin changes. Biopsy specimens were taken from four specified skin locations (lateral to bridge of nose, ventral midline, dorsal midline, perineal area) and from additional areas where skin lesions were present. Samples were routinely processed and evaluated by light microscopy. Blood samples were collected and tested for estrogen, thyroid-stimulating hormone, triiodothyronine thyroxine, and cortisol levels. The axilla was swabbed and samples were obtained for bacterial culturing. All nine of the geriatric monkeys had notable dermal lesions, while one of the control monkeys and one of the sun-exposed monkeys had abnormal findings. Major gross findings included increased areas of erythematous skin, wrinkling, focal skin scaling, thinning of hair, foot calluses, and exudative lesions. Histologic skin changes included subacute dermatitis, acanthotic dermatitis, and a lesion resembling an early solar lentigo in the sun-exposed animal. These changes were not associated with hormonal abnormalities or bacterial pathogens. Histologic changes are compatible with nonspecific skin changes observed in elderly humans. This study establishes a baseline of dermatologic changes of the aging rhesus macaque. PMID- 9210027 TI - Intestinal parasitism--protozoa and helminths--in primates at the Barcelona Zoo. AB - The faunistic results regarding intestinal parasitism by protozoa and helminths in 21 primate species (three Cebidae, thirteen Cercopithecidae, one Hylobatidae, one Lemuridae, three Pongidae) are reported. The primate species were housed in four separate galleries. Six faecal samples of each host species were subjected to coprological analysis. Fifteen parasite species were detected: 11 protozoa (Entamoeba coli, E. chattoni, E. hartmanni, Iodamoeba butschlii, Endolimax nana, Giardia intestinalis, Chilomastix mesnilii, Enteromonas hominis, Trichomonas intestinalis, Balantidium coli, and Blastocystis hominis) and 4 helminths (Ancylostoma sp., Strongyloides fuelleborni, Strongyloides sp., and Trichuris trichiura). The results reveal certain parasitic similarities between host species housed in the same gallery; however, these primate species do not always carry identical parasite species. PMID- 9210028 TI - Tumors of the respiratory tract observed at the German Primate Center, 1978-1994. AB - Eight spontaneous pulmonary tumors (four bronchiolar tubular adenomas, two bronchiolar adenocarcinomas, two squamous-cell carcinomas) occurred in a total of 54 adult tree shrews (Tupaia belangeri) of the GPC colonies between 1978 and 1994. The adenomas and adenocarcinomas consisted of tubularly or trabecularly arranged cuboidal to cylindrical cells interspersed with some PAS-positive goblet cells, thus resembling the epithelial lining of respiratory bronchioles of tree shrews. The two squamous-cell carcinomas probably originated from the pulmonary alveoles. Three more pulmonary tumors (one small-cell carcinoma, one bronchial adenoma, one squamous-cell carcinoma) developed in 409 adult callitrichids of the GPC colonies during the same period, and one more bronchial adenoma was observed in a common marmoset (Callithrix jacchus) of another colony located in Gottingen. With regard to the adenomas and squamous-cell carcinomas, a similar cellular origin with the three shrews is assumed. The small-cell carcinoma possibly developed from the bronchial epithelium, provided a pathogenesis parallel to that of human small-cell carcinoma is suggested. Four of the tree shrew pulmonary adenomas/adenocarcinomas and the small-cell Ca were macroscopically visible as yellowish-grey nodules of 1 mm x 1 mm to 15 mm x 15 mm diameter, predominantly involving the main lobes (2 x right main lobes, 2 x left main lobes, 1 x all lobes). The pulmonary tumors of the other animals were below macroscopical detectability. PMID- 9210029 TI - Colonic adenocarcinoma in a rhesus macaque (Macaca mulatta). AB - A spontaneous colonic adenocarcinoma and endometriosis was diagnosed in a 34-year old female rhesus macaque (Macaca mulatta). The tumor caused partial obstruction of the ascending colon and histologically resembled the commonly described napkin ring tumors of the descending and sigmoid colon found in humans. Serum levels of CA 125, a high-molecular-weight glycoprotein antigen that has been reported elevated in a variety of pathological conditions of the pelvic cavity in humans, was severely elevated. Both the adenocarcinoma and the endometriosis may have contributed to this finding. PMID- 9210030 TI - Comprehensive investigation of the presence of JC virus in AIDS patients with and without progressive multifocal leukoencephalopathy. AB - Progressive multifocal leukoencephalopathy (PML), a viral-induced demyelinating disease, is becoming relatively common, while many diagnostic and pathogenetic aspects remain to be clarified. A study was therefore undertaken in 64 AIDS patients suffering from various neurological disorders, including PML (12 subjects), with the specific objective of searching for JC virus (JCV) DNA by nested PCR (n-PCR) in cerebrospinal fluid (CSF), peripheral blood mononuclear cells (PBMCs), and urine collected from all patients. CSF examination, CD4 and CD8 counts, neurological examinations, and neuroradiological investigations were undertaken. JCV DNA was detected in 92% of CSF specimens in 75% of the PBMCs and urine samples from the PML patients, whereas among the non-PML patients JCV DNA was not detected in any CSF samples, but was found in 10% of PBMCs and in 39% of the urine specimens. BKV and JCV DNA viruria was observed simultaneously in 6% of the AIDS patients without PML. The routine CSF tests including IgG oligoclonal bands, the Link, and Tourtellotte IgG indexes, did not show a typical pattern in PML cases. The data obtained clearly indicate that the detection of JCV DNA in CSF constitutes an efficient marker for PML diagnosis. The simultaneous presence of JCV DNA in the CSF, PBMCs, and urine samples from the PML patients, who did not differ from controls with regard to their immunosuppressive status, suggests that JCV could be carried into the central nervous system (CNS) by infected PBMCs. PMID- 9210031 TI - Detection of archetype and rearranged variants of JC virus in multiple tissues from a pediatric PML patient. AB - JC virus (JCV) establishes persistent infections in its human host, and in some immunocompromised individuals, the virus causes the fatal brain disease progressive multifocal leukoencephalopathy (PML). Two forms of the virus, archetype and rearranged, have been isolated, with the latter being derived from the archetype form by deletion and duplication of sequences within the viral transcriptional control region (TCR). We have used the PCR technique to amplify JCV TCR sequences present within multiple tissues of a pediatric PML patient and have cloned and sequenced the PCR products. Archetype JCV was readily detected in kidney tissue; this form of JCV was also identified for the first time in brain and lymph node tissue by employing archetype-specific PCR primers. In addition, several archetype-like variants containing small deletions within their regulatory regions were isolated from cardiac muscle and lung. Different, but related rearranged forms were detected in most of the tissue examined. Each of the rearranged TCRs lacked portions of a 66 base pair (bp) region found within the archetype promoter-enhancer but retained a 23 bp region that is deleted in the prototype (Mad 1) rearranged form of JCV. Although several rearranged forms of JCV were identified in this patient, the TCRs could be assigned to one of two groups based upon the deletion boundaries generated during the adaptation from archetype to rearranged JCV. This study is the first to characterize multiple JCV variants present in different tissues from a patient likely to have succumbed to PML during a primary infection. PMID- 9210033 TI - Sites other than nucleotide 234 determine cardiovirulence in natural isolates of coxsackievirus B3. AB - The genetic site(s) that naturally determine the cardiovirulence phenotype of coxsackievirus B3 (CVB3) have yet to be mapped. Using two closely related CVB3 strains that differed in terms of cardiovirulence phenotype in mice, we previously reported the difference in phenotype mapped to a single site, nucleotide 234 (nt234) in the 5' non-translated region (NTR) of the CVB3 genome. When nt234 was C, the virus was attenuated and when U, the virus was cardiovirulent. To determine whether this finding was applicable to other strains of CVB3, we examined 13 different naturally occurring CVB3 strains isolated in different years in the United States. We determined that only two isolates induced severe inflammatory heart muscle disease in C3H/HeJ male mice. Using PCR products as sequencing templates, we determined the 5' NTR sequence from each viral genome. Alignment of these sequences and other published CVB3 5' NTR sequences suggests as many as four separate lineages, with commonly used laboratory strains clustering closely in one branch. An examination of the sequences showed that regardless of cardiovirulence phenotype, nt234 was invariably uridine. Thus, the previously reported cytidine at nt234 is most likely the result of a rare mutation and is not a naturally occurring variation and other sites must account for the variance in virulence seen in natural isolates of CVB3. PMID- 9210032 TI - Incidence of JC viruria is higher than that of BK viruria in Taiwan. AB - To investigate the prevalence of human polyomaviruses in Taiwan, urine samples from immunocompetent (healthy), transient immunocompromised (pregnant), and prolonged immunosuppressed (autoimmune disease) individuals were collected throughout the island. The viral DNA in the urine was detected by the polymerase chain reaction (PCR) and Southern blot. The viral genotypes were determined by DNA sequencing within the regulatory region. The overall results, including cases reported previously, show that 13.3% (10/75) of immunocompetent individuals, 26.0% (20/77) of pregnant women, and 37.5% (18/48) of autoimmune disease patients are JCV positive. All of the immunocompetent individuals are BKV negative, but 3.9% (3/77) of the pregnant women and 6.2% (3/48) of autoimmune disease patients are BKV positive. Twenty-four percent (48/200) of the examined urine samples were JCV positive, but only 3% (6/200) were BKV positive. JCV positive individuals were mainly infected with CY (42%) and TW-1 (52%) subtypes. These results suggest that the incidence of urinary excretion of human polyomaviruses in immunosuppressed individuals is higher than that of immunocompetent individuals. The prevalence of JCV appears to be higher than that of BKV in Taiwan. In addition, CY and TW-1 are the predominant subtypes of JCV prevalent in the Taiwanese population. PMID- 9210034 TI - Evaluation of multiple antibodies to Epstein-Barr virus as markers for detecting patients with nasopharyngeal carcinoma. AB - Five serological tests were assessed for their sensitivity for screening and early detection of nasopharyngeal carcinoma (NPC). The tests included the detection of antibodies to various gene products of EBV: viral capsid antigen (VCA) using an indirect immunofluorescence assay (FA), DNase using an activity neutralisation test (NT), Dnase using an enzyme-linked immunosorbent assay (ELISA), DNA polymerase (DP) using NT, and major DNA binding protein (MDBP) by ELISA. Sera from 100 NPC outpatients and 20 NPC patients, who were detected in a prospective study, were examined. The results showed that levels of antibody to DNase detected by ELISA and to DP detected by NT and the positivity rate for VCA by FA increased with NPC stage. More species of EBV antibody became detectable as NPC progressed. The detection of anti-MDBP antibody by ELISA was suitable for screening for NPC. Anti-DP antibody detected by NT was a valuable marker both for early detection and prognosis of NPC. Detection of anti-DNase antibody by ELISA was the most sensitive method for detection of NPC. No single test was sufficient to detect all the NPC patients and a combination of anti-DNase by ELISA with other tests are recommended to identify NPC patients. PMID- 9210035 TI - Positive and negative strand of hepatitis C virus RNA sequences in peripheral blood mononuclear cells in patients with chronic hepatitis C: no correlation with viral genotypes 1b, 2a, and 2b. AB - Hepatitis C virus (HCV) has many genotypes which are closely associated with the severity of chronic hepatitis and the response to antiviral therapy. Although HCV is essentially hepatotropic, several lines of evidence suggest that this virus can infect peripheral blood mononuclear cells (PBMC) in most patients with chronic HCV infection. However, the methods used previously to detect negative strand HCV RNA have been questioned, and the PBMC tropism of different HCV genotypes remains unknown. A stringent method was used to investigate the prevalence of positive- and negative-strand HCV RNA in the PBMC of 106 patients with chronic hepatitis C and to analyze the influence of HCV genotype on the tropism of PBMC. HCV type 1b was the predominant strain in the patients. Positive strand RNA in PBMC was detected in 83 (78%) and 40% had negative-strand RNA. The demographic and clinical features were comparable among different patients grouped by the replication status of HCV in the plasma and PBMC samples. In addition, there was no significant difference of PBMC tropism between type 1b and non-1b HCV. In summary, HCV does indeed infect actively the PBMC of chronic hepatitis C patients and such infection is not correlated to the pathogenesis of liver cell damage. Moreover, the genotype is not associated specifically with PBMC tropism of HCV. PMID- 9210036 TI - Enzyme-linked immunosorbent assay-IgG antibody avidity test for single sample serologic evaluation of measles vaccines. AB - A measles-specific enzyme-linked immunosorbent assay (ELISA)-IgG avidity test for serologic evaluation of the efficacy of measles vaccines with only one blood sample was evaluated after vaccination with three measles vaccine strains. Avidity indices were determined by the urea elution technique in samples presenting antibody titers > or = 100 mIU/ml. All 127 sera collected 2-8 weeks after primary vaccination with Biken-CAM70 measles vaccine had low avidity indices (LAI, when < or = 29%) with a time-dependent increase in avidity. In samples collected 6-10 weeks after vaccination with Edmonston-Zagreb, LAI were also observed in all 31 sera tested (mean = 15%) and in 233/242 (96.3%) filter paper samples from primary vaccination with Schwarz vaccine (mean = 14%). There was no difference in the mean avidity among the three groups of primary vaccinees, although the Schwarz group had higher antibody titers. In contrast, only 1/36 (2.8%) serum samples from children who were seropositive at the time of measles vaccination had LAI (mean = 56%), despite the fact that they were collected early (2-5 weeks after vaccination). Of 90 serum samples from children vaccinated in the past with two doses and of 42 cord blood serum samples, none had LAI. It is concluded that this test is a good tool for evaluating serologically the efficacy of a single dose schedule of measles vaccine. With only one postvaccination sample, the test can discriminate nonresponders (antibody titers below 100 mIU/ml), primary responders (antibody titers > or = 100 mIu/ml with LAI), and those previously immunized (antibody titers > or = 100 mIU/ml with high avidity indices). The seroconversion rate can be calculated after excluding the latter. PMID- 9210037 TI - A novel assay for hepatitis B e markers. AB - The evaluation of a novel assay that allows simultaneous testing for hepatitis B e antigen and its antibody in a single well is described. The results of routine application and sequential studies on patients with acute hepatitis B and chronic hepatitis B treated with interferon are presented. The specificity and sensitivity of the assay and its ability to be used to follow the response of a patient during the whole seroconversion episode has been evaluated. The assay proved to give useful information about the reactivity of the sample, especially in those patients who were changing their "e" status. PMID- 9210038 TI - Lack of susceptibility of the cottontop tamarin to hepatitis C infection. AB - The only species apart from man that is known to be susceptible to HCV infection is the chimpanzee but the availability of this primate for research is strictly limited. In an attempt to find an alternative and more practical model for HCV studies three cottontop tamarins were inoculated intravenously with HCV containing serum from patients with chronic HCV infection. The tamarins were monitored regularly for biochemical indications of hepatic inflammation and serum samples were assayed at weekly intervals for the presence of HCV-RNA and HCV antibodies. HCV-RNA was detectable at 10 minutes postinoculation in all three animals but not at any later time point over a 6 month period. No evidence of an active humoral immune response to the inoculated HCV was obtained although passively transferred anti-HCV was detectable in one animal until 1 week postinoculation. Biochemical findings did not indicate hepatic inflammation and liver histology remained normal. It is concluded on the basis of these negative findings that the cottontop tamarin is not susceptible to HCV infection. PMID- 9210039 TI - Amino-terminal epitopes are exposed when full-length open reading frame 2 of hepatitis E virus is expressed in Escherichia coli, but carboxy-terminal epitopes are masked. AB - We constructed a panel of overlapping and non-overlapping fragments of cDNA derived from open reading frame 2 (ORF2) of hepatitis E virus (HEV) and fused to the gene encoding glutathione S-transferase (GST), from which proteins were expressed in Escherichia coli. IgG-specific immunoreactivity against each protein was measured by Western immunoblotting using sera from experimentally infected Rhesus macaques (Macaca mulatta) or from HEV-infected patients. Under these conditions, full-length ORF2 protein (GST-ORF2) was strongly reactive with acute phase sera from either macaques or patients, but was poorly reactive with convalescent sera. Recombinant protein GST-ORF2.3, representing amino acids 1-110 of the 660 encoded by ORF2, demonstrated a pattern of reactivity largely indistinguishable from the full-length protein. Conversely, GST-ORF2.1, representing amino acids 394-660 of the ORF2 protein was strongly reactive with both acute- and convalescent-phase sera. Extension of GST-ORF2.1 towards the N terminus led to a progressive loss of convalescent-phase reactivity, apparent with as few as 20 additional HEV-specific amino acids. Deletion of 40 or more amino acids from the N-terminus of ORF2.1 also led to reduced convalescent-phase reactivity, however a protein representing this "reactive" region, containing amino acids 394-473, was poorly reactive, suggesting that the convalescent reactive epitopes are conformational. Expression of full-length ORF2 protein in E. coli therefore masks the convalescent-reactive epitopes within the C-terminal part of the protein, without affecting N-terminal, acute-reactive epitopes. PMID- 9210040 TI - Fulminant demyelinating encephalomyelitis associated with productive HHV-6 infection in an immunocompetent adult. AB - Human herpesvirus 6 (HHV-6), the etiologic agent of roseola in young children, has been reported to be detectable in the brain of many neurologically normal adults, although regional localization to plaques of multiple sclerosis has also been demonstrated. Large amounts of this virus were present in multifocal demyelinating white matter lesions of fulminant encephalomyelitis with seizures in a 21-year-old woman with normal immune parameters. Brain biopsy after 3 weeks of neurologic deterioration revealed a viral etiology by light and electron microscopy; the virus was identified as HHV-6 by immunohistochemistry and by polymerase chain reaction (PCR) amplification in biopsy and autopsy specimens. PMID- 9210041 TI - Temporal development and prognostic value of antibody response to the major neutralizing epitopes of gp120 during HIV-1 infection. AB - Our objective was to analyse the humoral response to the major neutralizing epitopes of gp120. The kinetics of the appearance of antibodies directed to the V3 region (V3 Abs) and antibodies directed to the CD4 binding site (CD4BS Abs) were compared in sequential sera from 20 seroconverters. V3 Abs were titrated using 2 different indirect EIAs with synthetic oligopeptides coated on the solid phase. The sequences of the oligopeptides used were those of the MN isolate or a mixture of the consensus sequences of the 5 major HIV-1 subtypes (A-E). CD4BS Abs titers were determined using an EIA in which serum antibodies compete with a labeled human monoclonal antibody, F105, whose corresponding epitope overlaps the conformation-dependent CD4BS, for binding to purified recombinant gp120 coated on a solid phase. The prognostic value of both antibodies was analyzed in a longitudinal study of 60 HIV-1 infected patients (17 nonprogressors and 43 progressors). Eighty-five percent and 70% of HIV sero-converters were positive for V3 Abs and CD4BS Abs, respectively, during the observation period. V3 Abs were detected first in the majority of the patients (mean delay of appearance, 1.22 +/- 0.96 months vs. 4.81 +/- 2.05 months for CD4BS Abs). Both categories of antibodies appeared simultaneously in 4 patients (20%). No prognostic value could be attributed to these antibodies. Our data confirm that V3 Abs and CD4BS Abs appear with some delay after primary infection, suggesting that they do not play a large or early role in the rapid clearance of viremia in primary HIV-1 infection. These antibodies were not associated with progression to symptomatic infection and are thus of no value for surveillance in HIV-1 infected patients. PMID- 9210042 TI - Detection of varicella-zoster virus DNA in patients with acute peripheral facial palsy by the polymerase chain reaction, and its use for early diagnosis of zoster sine herpete. AB - Varicella-zoster virus (VZV) reactivation without cutaneous vesicles (zoster sine herpete) has been demonstrated in 8 to 25% of patients with acute peripheral facial palsy (APFP) by serological methods. To make an early diagnosis of zoster sine herpete, VZV DNA in oropharyngeal swabs from patients with APFP was examined by the polymerase chain reaction (PCR). VZV DNA was detected in oropharyngeal swabs from 6 of 36 (17%) patients with APFP by PCR. VZV DNA was detected in the oropharyngeal swabs from the six patients at their initial visit (2 to 4 days after the onset of APFP), while the anti-VZV IgM and IgG antibody titers were not increased significantly. In contrast, VZV DNA was undetectable in the oropharyngeal swabs at the time when the VZV specific antibody response appeared. These results indicate that detection of VZV DNA in oropharyngeal swabs by PCR is more useful than currently available serological assays for the early diagnosis of zoster sine herpete in patients with APFP. PMID- 9210043 TI - Low rate shedding of HSV-1 DNA, but not of infectious virus from human donor corneae into culture media. AB - Fluid samples derived from 451 organ cultured corneae were tested for the presence of HSV-1 DNA after electroseparation and amplification for fragments of the glycoprotein D- and thymidine kinase-encoding genes. Of the culture media, 134 were processed immediately after withdrawal (Group 1); 100 were stored at ambient temperature for 6 to 60 weeks (Group 2); 90 were stored at -8 degrees C for 4 to 9 weeks (Group 3); and 127 were stored at -20 degrees C for 2 to 30 weeks (Group 4). The degradation of human DNA (marker gene, betaglobin) under these different storage conditions and of human and HSV-1 DNA as a sequential function of time at ambient temperature was gauged by the loss of a detectable signal for the respective component. Endothelial cell density within each of the corneal discs was determined before and after organ culture. In 7/451 culture fluid samples, HSV-1 DNA corresponding to either the glycoprotein D- or thymidine kinase-encoding genes was detected. In culture fluid samples derived from Group 2 at ambient temperature, for 6 to 60 weeks) and 3 (at -8 degrees C, for 4 to 9 weeks), complete degradation precluded the detection of human DNA, and hence probably also of HSV-1 DNA; only at -20 degrees C did DNA remain stable for protracted periods of time. Even so, HSV-1 DNA was detected in only 2% of those media in which no degradation was to be expected; additionally, there existed no correlation between its presence in culture fluid samples and the loss of endothelial cells or cytopathic changes. DNA can be extracted successfully and concentrated twenty-fold from high-volume samples by electroseparation. When shed into culture fluid, it is remarkably prone to a time and temperature dependent degradation, which may lead to false negative results. It is concluded that there is no infectious virus to be expected in the specimens; the occurrence of HSV-1 DNA in donor corneae would not appear to be an important factor influencing their biological quality during the period of organ culture. PMID- 9210044 TI - Prevalence of antibodies to HTLV in antenatal clinic attenders in south east London. AB - The prevalence of antibodies to HTLV in women attending a south east London antenatal clinic between October 1990 and July 1992 was determined using sera referred for routine rubella antibody testing. Samples were screened for HTLV antibody using a modified Fujirebio gel particle agglutination test and reactive sera confirmed by ELISA (Abbott Laboratories, North Chicago, IL) and two commercial Western blots (Cambridge Biotech Inc., Rockville, MD, and Diagnostic Biotechnology, Genelab Diagnostics, Louvaine, Belgium). This strategy confirmed the presence of HTLV-1 antibodies in 12 out of 6,289 sera (0.19%, 95% confidence limits 0.083% to 0.30%) and HTLV-2 antibodies in 2 (0.03%) sera. Specimens from 8 of 821 (0.97%, 95% confidence limits 0.42% to 1.9%) Afro-Caribbean women, three of 1,136 (0.26%, 95% confidence limits 0.055% to 0.78%) African women, and one of 3,049 (0.033%, 95% confidence limits 0.006% to 0.18%) Caucasian women were positive for HTLV-1 antibodies. Sera from Afro-Caribbean women born in the Caribbean were 7.6 times more likely to be HTLV-1 antibody positive than sera from Afro-Caribbean women born in the UK (P = 0.012). Selective testing of Afro Caribbean and African antenatal clinic attenders, in this setting, would have identified 11 of the 12 HTLV-1 infections at an estimated cost of prevention of HTLV-1 associated disease of 100,000 pounds per case which is considerably less than the 1.3 million pounds which has been estimated to prevent a case by universal screening of UK blood donors. PMID- 9210045 TI - Higher prevalence of Borna disease virus infection in blood donors living near thoroughbred horse farms. AB - It is believed that Borna disease virus (BDV), an etiological agent of progressive polioencephalomyelitis in horses and sheep, is closely associated with psychiatric disorders in humans since the prevalence of BDV is higher in psychiatric patients than in blood donors. We investigated whether or not BDVs in humans are derived from infected domestic animals, by characterizing the BDVs in blood donors and horses derived from the same region of Hokkaido island, Japan. The seroprevalences (2.6 to 14.8%) of BDV were significantly higher in the blood donors from four regions where most horse farms are concentrated, compared with only 1% in the blood donors from Sapporo, the largest city in Hokkaido.BDV RNA was also detected in peripheral blood mononuclear cells from most of the seropositive horses and blood donors by nested reverse transcriptase-polymerase chain reaction. These findings support that BDV may be horizontally transmitted, at least in part, from infected horses to humans. PMID- 9210046 TI - Are known pyrogenic cytokines responsible for fever in influenza? AB - The levels of interleukin (IL)-1 beta, IL-6, tumour necrosis factor (TNF)-alpha, and macrophage inflammatory protein (MIP)-1 alpha released from human peripheral blood leucocytes (PBL) following interaction with influenza virus clone 7a (virulent, produces high fever in ferrets) and A/Fiji (attenuated, produces relatively low fever in ferrets) were low and similar for the two viruses. Neither strain induced interferon (IFN)-gamma and release of IL-8 (which occurs on incubation of PBLs alone) was reduced after interaction with the two viruses. The levels of IL-1 and IL-6 detected in the plasma of infected ferrets were low and did not correlate with the onset, duration or magnitude of the fevers produced by clone 7a and A/Fiji. Relatively large amounts (100,000 pg/kg) of IL-1 and TNF-alpha were needed to produce appreciable fever in rabbits, and such quantities of IL-6 were not pyrogenic. Hence, as for previous observations, no evidence could be obtained that induction of known pyrogenic cytokines is responsible for the febrile response in influenza. The possibility that some other mediator(s) may be involved cannot be ruled out. PMID- 9210047 TI - Genomic regions of coxsackievirus B3 associated with cardiovirulence. AB - The molecular basis for cardiovirulence in the coxsackievirus B3 (CVB3) genome was examined in a murine model of acute myocarditis. Infectious cDNAs representing a highly cardiovirulent coxsackievirus B3 (CVB3m) and a noncardiovirulent (CVB30) virus were used to construct infectious chimeric cDNAs. Assays of the resulting recombinant viruses for cardiovirulence in adolescent male CD-1 mice showed that the 5' nontranslated region (5' NTR) of the CVB3m genome plays the major role in determining cardiovirulence and that the genomic region encoding the capsid proteins has a minor additive effect in increasing cardiovirulence. Nucleotide sequences in the 5' NTR of CVB3m and CVB30 differ at 23 positions; 14 are located in four stemloop motifs of the secondary structure and may influence the cardiovirulent phenotype by regulating RNA or protein synthesis. A comparison of predicted amino acid sequences of capsid proteins in CVB3m and CVB30 identified two amino acids as potential candidate contributors to cardiovirulence, i.e., amino acids at positions A207 (Asn-Asp) in the puff structure of the E-F loop of VP2 and A566 (Gln-Glu) in the C terminal of VP3 at the external surface. The data from this study and published literature support the conclusion that cardiovirulence of a CVB3 can depend on several regions of the genome. PMID- 9210048 TI - Low-grade gliomas: introduction and overview. AB - This issue of the Journal of Neuro-Oncology is devoted to recent investigations of low-grade gliomas. The purpose of this issue is not to debate the relative merits and liabilities of different management strategies for low-grade gliomas, but to present new data concerning novel and innovative approaches to evaluating these lesions. The common theme of many of these reports represents a departure from grading systems that primarily depend on a morphology-based analysis from light microscopy to classify these tumors. The purpose of this review is to present the reasoning behind the selection of authors for this issue of the Journal of Neuro-Oncology and to provide a format for presentation of new ideas concerning these interesting tumors. It is clear that standard classification systems that address only the morphological characteristics of tumor cells can not adequately represent the wide variation in biological activity that is found with these lesions. It is hoped that these articles will stimulate further interest and research into low-grade gliomas that will one day lead to more effective therapy. PMID- 9210050 TI - Early vs. delayed radiotherapy in a small cohort of patients with supratentorial low grade glioma. AB - In a cohort of 25 patients with supratentorial low grade glioma, the timing of radiotherapy made no significance difference for 10 year survival. Patients who received early radiotherapy had a 55% 10 year survival and those receiving delayed radiotherapy had a 53% 10 year survival. Radiotherapy was delayed a median of 4.8 years in those receiving delayed radiotherapy. PMID- 9210049 TI - Glial ontogeny and glial neoplasia: the search for closure. AB - The last ten years have seen rapid progress in both our understanding of the normal progression and control of gliogenesis and in the laboratory techniques necessary to sustain and study most glial cell types, including progenitor cells of both type-1 astrocyte (T1A) and oligodendrocyte-type-2 astrocyte (T2A) lineage. These studies have direct relevance for the lineage analysis of human gliomas, optimizing in vitro glioma models, and suggesting potentially fertile new grounds for glioma biology research. We do not yet known whether malignant transformation occurs only in mature glia that then 'de-differentiate' into cells with glial precursor phenotypes and behavior characteristics, whether neoplastic transformation occurs in O-2A progenitor cells, or whether both mechanisms may occur in different patients. However, preliminary results suggest that astrocytomas can arise from two different glial lineages, that oligodendrogliomas and mixed oligo-astrocytomas arise exclusively from the O-2A lineage, and that medulloblastomas may also have a connection with the O-2A lineage. An ontogeny based glioma classification system may lead to better prognostic patient data and better predict patient response to treatment than currently available classification systems. Available data from the study of developmental glial biology raises serious doubts about the fidelity and relevance of in vitro glioma models that rely on culture media supplemented with animal serum and suggest that relying on chemically-defined media conditioned by astrocytes may be the better research strategy. Findings from the study of normal gliogenesis also suggest that growth factors are likely to act as much more than simple mitogens in glioma biology. Potentially fertile areas of research for glioma biology include studying the cooperative effect of multiple growth factors, potential growth factor effects as survival factors, inhibitors of differentiation, and differentiation inducers, and studying potential positive humoral feedback loops between glioma cells and normal glial cells, as well as normal non-glial cells, within and surrounding each glioma. PMID- 9210051 TI - Proliferative activity and prognosis of low-grade astrocytomas. AB - Well-differentiated astrocytomas may transform into malignant astrocytomas in time. In surgical specimens, when the histological picture strictly corresponds to that of grade II glioma, the transformation is unpredictable. Clinically, the bad outcome of a quota of astrocytomas is a well known phenomenon. The use of proliferation markers, and recently of MIB-1 LI, for detecting the proliferation potential comes out to be a useful tool for prognosis. A survival analysis of fifty astrocytomas grade II according to the WHO classification was performed with univariate and multivariate analysis of a series of clinical and histological parameters. MIB-1 LI was calculated and compared with all the other parameters. A cut-off of 8% of MIB-1 LI divided the astrocytomas in two groups with significantly different survival (p = 0.0066): median survival time of 1062 versus 1686 days. According to multivariate analysis MIB-1 LI resulted to be an independent factor (p = 0.002) along with extension of surgical removal (partial versus total), postoperative Karnofsky status (> or = 70 versus < 70) and age (< or = 30 versus > 30). The interpretation of well-differentiated astrocytomas with high MIB-1 LI is that the increasing number of cycling cells precedes phenotypic transformation. MIB-1 LI can be used as a prognostic factor. PMID- 9210052 TI - Oligodendrogliomas. Part I: Patterns of growth, histological diagnosis, clinical and imaging correlations: a study of 153 cases. AB - The present study has attempted to demonstrate that the morphological spectrum of oligodendrogliomas includes tumors which are traditionally misinterpreted as 'diffuse fibrillary astrocytoma'. We have shown that these tumors are in fact made of isolated neoplastic oligodendrocytes which are entrapped in a fibrillary background composed of axons and fibrillary reactive gliosis. Analysis in a series of 153 'pure' supratentorial oligodendrogliomas composed of 'classical' or pseudo 'diffuse fibrillary oligodendrogliomas' diagnosed by imaging-based serial stereotactic biopsies showed that 2/3 of the tumors were exclusively made of isolated tumor cells (ITCs) (structure type III) and that only 1/3 of them exhibited both ITCs and solid tumor tissue components (structure type II). The tumor tissue destroys the brain parenchyma and contains new formed microblood vessels whereas ITCs do not destroy the parenchyma and are not associated with microangiogenesis. These fundamentally opposite morphological characteristics were reflected by the following findings: 1) contrast enhancement was observed in 64% of structure type II but was never seen in structure type III oligodendrogliomas. 2) a neurological deficit occurred in 57% of structure type II but in only 8% of structure type III oligodendrogliomas. 3) using the new grading system described in the companion paper to this study, we found that the biological behavior of oligodendrogliomas was also closely related to the patterns of tumor growth. From a synthesis of data gathered in this study it is suggested that emergence of microangiogenesis within a tumor which at first grows slowly with a structure type III pattern is a crucial event toward more aggressive behavior. PMID- 9210053 TI - Oligodendrogliomas. Part II: A new grading system based on morphological and imaging criteria. AB - This second part of our study of 'pure' oligodendrogliomas focuses on survival data analysis. In order to identify potentially useful prognostic factors and to assess the effectiveness of a new grading system, the 79 patients in the previously analyzed series for whom adequate follow-up could be obtained (52%) were entered in the present analysis. Statistical analysis demonstrated that contrast enhancement and endothelial hyperplasia had powerful and similar influence on survival. Median survival with and without contrast enhancement were: 3 versus 11 years, and with or without endothelial hyperplasia were: 3.5 versus 11 years. Conversely, the degree of nuclear atypia and presence or absence of mitosis or necrosis were not correlated with survival. These findings allowed us to devise a simple grading system which discriminates two malignancy grades as follows: absence of endothelial hyperplasia and of contrast enhancement = Grade A, presence of endothelial hyperplasia and/or of contrast enhancement = Grade B. Of the 79 oligodendrogliomas in this study, 59 tumors were categorized as grade A and 20 as grade B. Median survival were: 11 years in grade A and 3.5 years in grade B. Five-year and 8-year survival rates were: 89% and 60% in grade A and: 33% and 15% in grade B. Double blind grading between two independent observers was concordant in 96% of the cases. Application of this simple efficient and reproducible grading scheme should permit reliable comparison of retrospective or prospective therapeutic data emanating from various institutions. PMID- 9210054 TI - Low-grade gliomas of chronic epilepsy: a distinct clinical and pathological entity. AB - The authors present a summary of their recent experience regarding the management of patients with a variety of low-grade gliomas found during the evaluation for chronic epilepsy. These tumors are notable because the long-term patient outcome in this population is significantly better than the anticipated results of patients with the same tumors who do not have chronic epilepsy. Based on the long history of preoperative seizures (median 14 years), the frequent cortical location, and the absence of tumor recurrence or anaplastic transformation and the lack of mortality in this population, low-grade gliomas of chronic epilepsy appear to define a specific pathological entity that separates them from other histologically similar low-grade gliomas. Low-grade gliomas of chronic epilepsy also are notable for the absence of morphological features that characterize with dysembryoplastic neuroepithelial tumors (DNTs). Our evidence suggests that low grade gliomas of chronic epilepsy should be recognized as a distinct pathological entity. PMID- 9210055 TI - Low grade gliomas: functional mapping resection strategies, extent of resection, and outcome. AB - The impact of surgery on outcome of adult patients with low-grade gliomas is controversial. Without prospective randomized treatment trials, one is primarily dependent on retrospective studies to address this issue. This paper reviews the recent clinical series of low-grade gliomas in which the association between extent of resection (EOR) and outcome could be analyzed. Functional stimulation mapping methods will be described to point out their critical role in maximizing the extent of resection while minimizing the risk associated with radical resection of low-grade gliomas. Studies from the CT-era analyzed with multivariate statistical methods were emphasized. The analysis of these studies points out that, for astrocytomas, there is no clear consensus that a greater EOR improves survival, but in most series under review, greater EOR significantly extended the survival of patients with oligodendroglioma. Unfortunately, there is little data which specifically analyzes and stratifies the outcome for other end points such as time to progression, malignant degeneration, mortality and morbidity, and duration of high quality survival by EOR. PMID- 9210056 TI - Expression of epithelial cadherin and cavernous sinus invasion in human pituitary adenomas. AB - Pituitary adenomas generally are regarded as benign tumors, although some invade the cavernous sinus and recur. We examined the epithelial cadherin (E-CD) expression in 30 pituitary adenomas (6 with cavernous sinus invasion and 24 without). Immunoreactivity of E-CD were found in all pituitary adenomas but they were very various. The presence of an association between E-CD expression and cavernous sinus invasion was assessed. There were no significant differences in E CD expression between invasive and noninvasive adenomas. These results suggest that E-CD expression is not associated with cavernous sinus invasion in pituitary adenomas. PMID- 9210057 TI - Lectin histochemistry of astrocytic tumors and in vitro characterization of lectin-induced modifications on the proliferation of the SW1088, U373 and U87 human astrocytic cell lines. AB - The role of lectins as biosignalling molecules or as markers of human astrocytic tumors remains relatively unexplored. The aim of the present work is to investigate (1) whether or not human astrocytic tumors express specific glycans, evidenced experimentally by means of lectin histochemistry, and (2) whether, in turn, these lectins can significantly modulate astrocytic tumor cell proliferation. Using a cell image processor, we therefore began by quantitatively measuring the histochemical binding pattern of 5 lectins (WGA, PNA, PHA-L, GSA IA4 and Con A) in 5 astrocytomas, 5 anaplastic astrocytomas and 5 glioblastomas. Secondly, we measured the influence of these 5 lectins on the proliferation of 3 astrocytic tumor cell lines (SW1088, U373 and U87) growing in vitro as monolayers. Cell proliferation was assessed by means of the colorimetric MTT assay. The histochemical lectin staining markedly varied intra- and inter-group. However, some constant results were obtained. Indeed, the staining increased markedly from GSA-IA4 and PHA-L through WGA and PNA to ConA in the three histopathological groups. The assessment of cell proliferation demonstrated that WGA, Con A and PHA-L very significantly decreased proliferation in the 3 astrocytic cell lines in a dose-dependent manner. Astrocytic tumor cells in the confluent growth phase were less sensitive to the WGA, Con A and PHA-L lectin induced effects than cells in the log growth phase. The GSA-IA4 and PNA lectins had globally very weak effects on the proliferation of the astrocytic tumor cell lines. Increasing the fetal calf serum from 1% to 10% in the culture media significantly antagonized the WGA-, Con A- and PHA-L-induced cell proliferation decrease in the 3 astrocytic cell lines. In conclusion, the present data strongly suggest that some lectins (including WGA, Con A and PHA-L) significantly influence the proliferation of astrocytic tumor cells. PMID- 9210058 TI - Intracarotid recombinant human tumor necrosis factor-alpha reduces cerebral blood flow and methionine uptake in rat brain tumors. AB - The aim of the present study is to understand the therapeutic effects of recombinant human tumor necrosis factor-alpha (rH-TNF) on hemocirculation and metabolism of brain tumors. Using double-label autoradiographic technique, we have monitored changes in regional cerebral blood flow (rCBF) and protein-bound fraction of (3H-methyl)-L-methionine, expressed as acid-insoluble fraction (AIF), in rat brain tumors following treatment with intracarotid rH-TNF. The central portion of tumors showed a significant decrease in rCBF and AIF at 4 hours after the injection (p < 0.01, p < 0.05, respectively, as compared with non-treated control rats), turned microscopically necrotic at 24 hours, and became more extensively necrotic at 72 hours. Tumor cells remained viable only in the peripheral portion of the tumors after the treatment. The peripheral portion also showed a moderate decrease in rCBF, but less change in AIF to 4 to 72 hours after the treatment. Neither ipsilateral nor contralateral non-involved cortex demonstrated appreciable changes in rCBF and AIF during the observed period. Intracarotid rH-TNF selectively reduces tumor rCBF and AIF, resulting in histological modification. PMID- 9210059 TI - Steroids decrease uptake of carboplatin in rat gliomas--uptake improved by intracarotid infusion of bradykinin analog, RMP-7. AB - A blood-tumor barrier (BTB) limits delivery of antitumor agents to brain tumors. This study sought to determine whether dexamethasone (DXN) treatment of rats with intracranial gliomas would 1) further impair delivery of carboplatin to brain tumors, and 2) whether intracarotid infusion of the bradykinin analog, RMP-7, would improve delivery during concurrent DXN treatment. Wistar rats with RG2 gliomas were utilized and a unidirectional transport, Ki, of radiolabeled [14C] carboplatin was determined using quantitative autoradiography. In DXN pretreatment animals, 3 mg/kg/day of DXN was administered intraperitoneally for 3 days prior to Ki determinations. At 10 days after tumor implantation, Ki of [14C] carboplatin into DXN-treated tumors and brain surrounding tumor (BST) was significantly lower compared to non-DXN treated tumors and BST (3.30 +/- 0.91 vs. 4.47 +/- 1.80, p < 0.05, and 0.94 +/- 0.84 vs. 2.18 +/- 0.79, p < 0.05, respectively). Intracarotid infusion of RMP-7 (0.1 mg/kg/min) significantly increased the Ki for carboplatin in DXN-treated tumors (6.35 +/- 3.10 vs. 3.30 +/ 0.91, p < 0.01), however, RMP-7 increased Ki to a greater extent in tumors not pretreated with DXN (12.07 +/- 3.60 vs. 4.47 +/- 1.80, p < 0.0001). Our studies show that dexamethasone decreases transport of carboplatin into brain tumors. Intracarotid infusion of RMP-7 selectively increases carboplatin transport to tumors. PMID- 9210060 TI - Adeno-associated viral vector gene transfer into leptomeningeal xenografts. AB - Leptomeningeal carcinomatosis is a painful and debilitating complication of cancer. Indwelling reservoirs provide continuous assess to the subarachnoid space, making leptomeningeal cancer potentially amenable to gene therapy. Adeno associated virus (AAV) is a defective virus not associated with any human disease. We used an AAV vector to transduce medulloblastoma (DAOY) cells in a nude rat model of leptomeningeal disease. After intraventricular injection of vector carrying the bacterial lacZ gene, beta-galactosidase positive cells were found in the implanted tumor and in ependymal and subependymal cells but not in underlying normal brain parenchyma. No evidence of virally-mediated toxicity was noted in the animals. The results of this pilot study demonstrate that AAV vectors may be used to transfer and express foreign genes in established leptomeningeal tumors. PMID- 9210062 TI - Recurrence from filum terminale ependymoma 42 years after 'total' removal and radiotherapy. AB - The authors describe the unusual case of a patient requiring reoperation for ependymoma of the filum terminale after a symptom-free period of 42 years: treatment of the primary tumor consisted of macroscopically complete surgical excision and postoperative radiotherapy. This case brings to light some interesting observations: a late recurrence, as in the case reported here, appears to be rare especially if the tumor presents in adult age; the long preoperative clinical history (26 months) and the initial aspect of the tumor that infiltrated 2 caudal roots, removed together with the tumor, may have been factors correlated with the risk of recurrence: postoperative radiotherapy may have helped to delay clinical manifestation of the recurrence. Ependymomas of the filum terminale may clinically recur even after complete removal. The latest clinical recurrence described in the literature occurred after 26 years. PMID- 9210061 TI - The treatment of recurrent cerebral gliomas with all-trans-retinoic acid (tretinoin). AB - Malignant gliomas continue to be a significant source of mortality in young and middle aged adults. The introduction of new treatment strategies and multidisciplinary approaches has improved the outcome of patients with these tumors only slightly. Because retinoic acid has growth inhibitory activity against glioma and neuroblastoma cells in cultures, we assessed the efficacy of all-trans-retinoic acid in the treatment of recurrent cerebral gliomas. Thirty six patients with recurrent cerebral gliomas were entered in the study and treated with 120 or 150 mg/ m2/day of all-trans-retinoic acid as a single agent. The drug was given for 3 weeks followed with one week of rest. Two blocks of 4 weeks constituted one course of treatment. One (3%) of 34 evaluable patients had a minor response and 14 (41%) had stable disease. In the rest of the patients (56%), tumors continued to progress despite treatment. The median time to progression of all evaluable patients was 8 weeks, and for the responders was 17 weeks. The higher dose level (150 mg/m2) was associated with high incidence of headache, which responded to dose reduction. The lower dose level was very well tolerated, with mild, mainly dermatological toxicity. All-trans-retinoic acid as a single agent has no significant activity against recurrent cerebral gliomas. PMID- 9210063 TI - Primary central nervous system lymphoma of T-cell origin: description of two cases and review of the literature. AB - Primary lymphomas of the central nervous system (PLCNS) of T-cell lineage are unusual. It has been suggested that T-cell PLCNS, compared to those of B-cell origin, present some differences in relation to age of presentation, gender, location of the tumor and survival. We describe two cases with T-cell PLCNS and review 22 parenchymatous T-cell PLCNS reported in the English literature. Age, gender and survival of the whole series of 24 T-cell PLCNS did not differ from that reported in large series of PLCNS where the great majority were of B-cell origin. In contrast, a location in the posterior fossa was found in 54% of T-cell PLCNS, whereas this location ranged from 12 go 29% in series of, mostly B-cell, PLCNS. T-cell PLCNS had a higher frequency (33%) of the histologic low grade small lymphocytic lymphoma than B-cell PLCNS (5%). Analysis of six T-cell PLCNS long-term survivors showed that half of them had low grade lymphomas. We conclude that T-cell PLCNS do not differ from those of B-cell origin in age of presentation or gender, but they have a preference to develop in the posterior fossa and a higher frequency of low grade histology which would probably explain the longer survival in some patients. PMID- 9210064 TI - Focal dystonia after chemotherapy: a case series. AB - Dystonia is a rare neurologic disorder of the basal ganglia presenting with involuntary twisting or turning spasm of muscles. Movements localized to the face, eyes, or neck generally present during late adulthood. Cranial dystonia is usually idiopathic but may be caused by trauma or medications. Of 148 patients with focal dystonia referred to Indiana University over four years, four women had the onset of face and neck symptoms eight days to 34 months after completing treatment with chemotherapy alone or combined with radiation therapy. Two patients were treated with 5-FU, one received doxorubicin and one was treated with both. Both drugs have been associated with transient parkinsonism, but no chemotherapeutic medications have been reported to cause dystonia. Three patients remain free of demonstrable malignancy. A possible association of chemotherapy and focal dystonia has not been previously described. PMID- 9210067 TI - Verbally administered Barthel Index as functional assessment in brain tumour patients. AB - OBJECTIVE: To investigate verbally administered Barthel Index as a measure of functional status in patients with high grade gliomas. BACKGROUND: Barthel Index (BI) is a performance score of activities of daily living which has been validated in patients with neurological disability. While any assessment of quality of life in brain tumour patients should include all the aspects of CNS function we concentrated on measurement of physical performance status and evaluated the role of BI as a measure of palliative effect of treatment in patients with high grade glioma undergoing radiotherapy. METHODS: BI was verbally administered on 504 occasions in 107 patients with high grade glioma. The BI scores were correlated with Karnofsky performance score (KPS), and neurological performance score (NPS) as a measure of inter-index reliability. The BI's prognostic value was assessed using actuarial survival data. RESULTS: BI was sensitive to change and reflected the degree of functional impairment. In patients with high grade glioma BI correlated with KPS, and NPS (R2 = 0.872 and 0.658 respectively). BI score was also of prognostic value in terms of survival. The median survival of patients with functional independence was 9 months with moderate disability 5 months and with severe disability 4 months. CONCLUSION: Verbally administered Barthel Index is easy to use, reliable and sensitive to change and is of prognostic value. It is a useful tool in the management of patients with gliomas, as an objective evaluation of palliative effectiveness of treatment in patients with functional disability. PMID- 9210065 TI - All-trans retinoic acid in relapsing malignant gliomas: clinical and radiological stabilization associated with the appearance of intratumoral calcifications. AB - PURPOSE: To evaluate the therapeutic effect of all-trans retinoic acid (ATRA) with and without cytosine arabinoside in relapsing malignant gliomas. PATIENTS AND METHODS: 9 patients (8 male, 1 female, age 53.9 +/- 11.2) with relapsing malignant gliomas (grade IV:6; grade III:3) were treated by ATRA 1 to 21 months after the end of their initial treatment. ATRA was given unceasingly during 2 to 17 months at 90 mg/d. In 6 patients it was associated to cytosine arabinoside (4 g/course, 1 to 9 courses every 4 weeks). RESULTS: 4 non-responder patients died 2.5 to 4 months after starting therapy. One patient who had been reoperated before receiving ATRA and cytosine arabinoside (5 course) had no sign of tumor recurrence after 17 months of treatment. In 4 responder patients (2 glioblastoma and 2 anaplastic astrocytoma) a clinical and radiological stabilization (time to progression) during 9 +/- 2.5 months was observed. This stabilization was associated in 3 of them with the appearance of intra tumoral calcifications visualized on repeated CT scans and confirmed in one patient by post-mortem examination. All of them had received cytosine arabinoside (1 to 9 courses) with ATRA; however small calcifications were also observed in one non-responder patient who did not receive aracytine. CONCLUSION: These results suggest: a) a therapeutic effect of ATRA in combination with cytosine arabinoside in patients with relapsing malignant gliomas b) that intratumoral calcifications are related to the effects of ATRA on differentiation and/or on endothelial t-PA production and that these effects explain the tumor progression arrest in responder patients. The transient efficiency is probably related to the pharmacokinetics of ATRA or to changes of cellular mechanisms that modulate the cell response to the drug and is a critical issue for this therapy. PMID- 9210066 TI - Advantage of treating anaplastic gliomas with aggressive protocol combining chemotherapy and radiotherapy. AB - We devised a treatment protocol for anaplastic gliomas consisting of:(a) chemotherapy prior to radiotherapy (b) a second chemotherapy regimen at tumor recurrence (c) repeated surgery whenever possible. 41 Anaplastic Astrocytoma (AA), 16 Anaplastic oligoastrocytoma (AOA) and 14 anaplastic oligodendroglioma (AOD) patients were treated. After surgery all patients received 5-6 cycles of carmustine+Cisplatinum chemotherapy. Radiotherapy was started during the last 2-3 cycles of chemotherapy. 17 patients (30.5%) were reoperated on at recurrence. All recurring patients underwent PVC chemotherapy. At this moment disease recurred in 56 patients. Median TTP was 24.5, 38.7 and 58.2 months for AA, AOA and AOD respectively. Median ST was 38.8, 71.8 and 73 months. In conclusion our sandwich protocol of prior chemotherapy, overlapping irradiation with second chemotherapy and, in favourable cases, a second surgical intervention, is of benefit in patients with anaplastic gliomas. PMID- 9210068 TI - All-trans-retinoic acid: a phase II Radiation Therapy Oncology Group study (RTOG 91-13) in patients with recurrent malignant astrocytoma. AB - The Radiation Therapy Oncology Group enrolled 30 patients with recurrent malignant astrocytomas onto a phase II study (RTOG 91-13). Patients were treated with all-trans-retinoic acid at a starting dose of 120 mg/m2 per day orally continuously until disease progression. Fourteen patients had glioblastoma, 14 had anaplastic astrocytoma, and 2 had other histologies; 53% were under 50 years of age. All patients had failed radiation therapy and/or at least one chemotherapy regimen. All patients had a Karnofsky performance status score of at least 70, but only 37% had a KPS of 90-100. Forty percent had a neurologic function status of grade 1 (able to work). A minimum of 4 weeks of all-trans retinoic acid defined adequate treatment. Twenty-five patients received adequate therapy. Most common toxicities were dry skin, cheilitis, anemia, and headache; 3 patients had grade 3 headache requiring suspension of all-trans-retinoic acid. No grade 3 hematologic toxicity was observed. Of 25 adequately treated patients, 3 showed objective regression of tumor on magnetic resonance imaging and computed tomography scans, 3 patients remained stable, and 19 patients had disease progression. The median time to tumor progression was 3.8 months and the median survival time was 5.7 months. This study suggests that this dose of single agent all-trans-retinoic acid has modest clinical activity against recurrent malignant gliomas with tolerable side effects. A response rate of 12% and a stabilization rate of 12% are lower than expected. Future studies with higher dosage or in combination with biological response modifiers or chemotherapy may be warranted. PMID- 9210069 TI - The doctor glut: I'm in a rut. PMID- 9210070 TI - Intrauterine cocaine exposure and the risk for sudden infant death syndrome: a meta-analysis. AB - OBJECTIVE: To determine whether an association exists between intrauterine cocaine exposure and sudden infant death syndrome (SIDS). STUDY DESIGN: A meta analysis of 10 published studies that reported the incidence of SIDS in infants born to mothers who used cocaine during pregnancy. We computed the rates of SIDS in cocaine-exposed infants for each of the 10 studies and in comparison groups for each of the eight studies that reported the incidence of sudden infant death syndrome in comparison groups. We also computed odds ratios and 95% confidence intervals individually for the eight studies with comparison groups. Then we computed adjusted overall odds ratios (Mantel-Haenszel) by combining the eight studies with comparison groups; combining three of these studies in which the comparison group infants were exposed to illicit drugs other than cocaine (polydrug controls); and combining the other five studies in which the comparison group infants were not exposed to illicit drugs (drug-free controls). RESULTS: For all 10 studies combined, SIDS occurred in 84 of 12,163 infants with intrauterine cocaine exposure; the variance weighted estimate for incidence was 5.2 per 1000 with a 95% confidence interval of 4 and 7 per 1000. The combined odds ratio for SIDS in cocaine-exposed versus all comparison group infants was 3.9 (95% confidence interval 3 and 5), a significantly increased risk. However, in a subset analysis we found that the odds ratio for SIDS in cocaine-exposed versus the polydrug comparison group infants was 2.7 (95% confidence interval 0.9 and 8.2), not a significantly increased risk. Whereas in a subset analysis comparing risk for SIDS in cocaine-exposed versus the drug-free comparison group infants, the odds ratio was 4.1 (95% confidence interval 3.2 and 5.3), indicating a highly significant effect of cocaine exposure on SIDS risk. CONCLUSION: After we controlled for the confounding variable of concurrent use of other drugs, the increased risk for SIDS could not be attributed to intrauterine cocaine exposure alone. The increase in risk for SIDS was found not to be specific to cocaine but to intrauterine exposure to illicit drugs in general. PMID- 9210071 TI - The single ventricle heart in the fetus: accuracy of prenatal diagnosis and outcome. AB - OBJECTIVES: The purpose of this study was to determine the diagnostic accuracy of fetal echocardiography in evaluating anatomic details of the single ventricle heart and the outcome of fetuses diagnosed with this anomaly. STUDY DESIGN: This is a retrospective study of 57 fetuses in which the results of fetal echocardiography were compared with the diagnoses at postnatal echocardiography, and postnatal surgical outcome was reviewed. RESULTS: Diagnostic accuracy was present in predicting morphology of the predominant ventricle, visceral situs, presence of pulmonary or aortic outflow tract obstruction, and presence of obstructed pulmonary venous outflow (sensitivity 100%). However, the ability to predict for a ductal dependent pulmonary circulation was poor (sensitivity 63%). Errors were made in the fetal assessment of ventricular size and viability such that in three cases, postnatal plans were altered toward a two-ventricular intervention. Of the 57 fetuses, intervention was elected in 37 (75%). Termination or nonintervention was elected in 14, and and 6 died before intervention. Of those operated on, 71% are presently alive after various stages of intervention. CONCLUSIONS: Accurate diagnosis of the fetal single ventricle heart is possible, and outcome is improving. Caution must be used in judging ventricular size and in predicting ductal dependent pulmonary circulation. PMID- 9210072 TI - Hemopericardium from coronary artery laceration complicating extracorporeal membrane oxygenation. AB - OBJECTIVE: We report the clinical course and successful surgical treatment of hemopericardium resulting from coronary artery (CA) laceration in two patients with congenital diaphragmatic hernia (CDH) undergoing extracorporeal membrane oxygenation (ECMO) bypass. STUDY DESIGN: Retrospective case review. RESULTS: Two neonates with CDH had needle aspiration for either pneumothorax or pericardial effusion before initiation of ECMO. While on bypass, progressive hemopericardium led to narrow pulse pressure and decreased venous return that limited bypass flow. Widened cardiac silhouette on chest radiographs suggested hemopericardium; echocardiography was confirmatory in one case. The underlying diagnosis of CA laceration was made during pericardiotomy and treated with surgical patching. CONCLUSIONS: Pre-ECMO history of cardiothoracic needle aspiration is important because complications such as hemothorax or hemopericardium may arise once ECMO bypass is initiated. Inadvertent CA laceration may lead to acute hemopericardium, compromising venous drainage. However, CA laceration can be successfully repaired while the patient is on bypass. PMID- 9210073 TI - Infant physiologic and behavioral organization during swaddled versus unswaddled weighing. AB - OBJECTIVE: The objective of this study was to evaluate the use of blanket swaddling during weighing on the physiologic and behavioral responses of preterm infants. STUDY DESIGN: A repeated-measures crossover design was used in a children's hospital intensive care nursery setting. Fourteen preterm infants with a mean gestational age of 32 weeks and mean weight of 1427 gm were weighed on two consecutive nights. A random start approach was used to determine initial order of swaddled or unswaddled weights. Effects of swaddled and unswaddled weighing were examined with use of the Assessment of Behavioral Systems Organization scales. Repeated-measures analysis of variance was used for analysis. RESULTS: Contrasts done on significant treatment-period interactions indicated that when swaddled, infants showed less physiologic distress (p < 0.002), better motor organization (p < 0.001), and more effective self-regulatory ability (p < 0.037) than when unswaddled during weighing. CONCLUSIONS: Swaddled weighing may decrease physiologic and behavioral distress in preterm infants. PMID- 9210074 TI - Tuberculosis skin testing in pregnancy: trends in a population. AB - BACKGROUND: A change has recently been noted in the epidemiology of tuberculosis in the United States. Multiple factors, including human immunodeficiency virus (HIV) infection, increases in the homeless population, and immigration, are cited as causes for an increased prevalence. The population of pregnant women in New Orleans exhibits several of these risk factors and may be compared with a previous description of this group reported in 1983. METHODS: All patients requesting obstetric care at the Medical Center of Louisiana at New Orleans from January 1994 to April 1995, were offered tuberculosis skin testing during their initial outpatient clinic visit. A skin test was considered positive if there was 10 mm of induration (5 mm in HIV-positive patients) at 48 hours. Other information collected included HIV status and ethnic group. This group was compared with a group of patients tested in 1981 and 1982. RESULTS: The study included 1621 patients, who underwent testing and had available results. The only significant risk factor for a positive skin test in 1994 was Hispanic ethnicity. Hispanics are the most recent immigrants to the New Orleans area. In 1983 Asians were at highest risk and were the newest immigrants to the city. HIV status was insignificant as a predictor of skin test conversion. CONCLUSIONS: Pregnant women at highest risk for tuberculosis in this urban center are recent immigrants to the United States. This is consistent with data reported by the Centers for Disease Control and Prevention from other locales. Efforts should be made to ensure that recently immigrated pregnant women receive skin testing as part of their obstetric care. PMID- 9210075 TI - Molecular zygosity studies aid in the management of discordant multiple gestations. AB - OBJECTIVE: Management of multiple gestations often requires identification of chorionicity. Sonographic evidence of dichorionicity can be present or inconclusive. DNA determination of zygosity can aid in the subsequent management of twins who are discordant for growth or anomalies. STUDY DESIGN: We present four cases in which monochorionicity could not be excluded sonographically. DNA zygosity studies were performed on amniocytes to guide management of the pregnancies. RESULTS: In two cases, one twin was affected with a significant anomaly. DNA zygosity studies provided a greater than 99% likelihood of monozygous twins in both cases. This altered the counseling regarding selective termination options. The other two cases involved severe intrauterine growth restriction and oligohydramnios in one twin, with a normal co-twin. The small twin appeared nonviable, and DNA studies were used to assess risk to the normal twin in the event of the co-twin's demise. We recommended delivery in the event of monozygous twinning and expectant management if the twins were dizygous. CONCLUSION: Diagnosis of chorionicity is important in multiple gestations. When chorionicity is unclear and management decisions would be altered by determination of zygosity, DNA studies should be considered. PMID- 9210076 TI - Intraventricular hemorrhage and fetal heart rate in very low birth weight infants. AB - OBJECTIVE: This study was designed to investigate the relationship between fetal heart rate patterns before delivery and periventricular-intraventricular hemorrhage in the very low birth weight infant. STUDY DESIGN: The last 30 minutes of electronic fetal heart rate data preceding delivery were analyzed for 84 singleton infants weighing between 700 and 1500 gm. All these infants received serial cranial ultrasonographic examinations commencing within 24 to 48 hours of birth. RESULTS: Thirty-three fetuses had normal heart rate patterns, and 51 had fetal heart rate abnormalities. Periventricular-intraventricular hemorrhage was not associated with fetal heart rate abnormalities. Univariate and multivariate regression analysis demonstrated that only gestational age < 28 weeks was a significant contributing factor to periventricular-intraventricular hemorrhage (odds ratio 2.2, 95% confidence interval [CI], 1.0 to 4.8). CONCLUSION: Fetal heart rate patterns immediately preceding delivery are not predictive of periventricular-intraventricular hemorrhage in the very low birth weight infant. PMID- 9210077 TI - Skin-to-skin holding in the neonatal intensive care unit influences maternal milk volume. AB - OBJECTIVES: To evaluate the effect of early initiation of skin-to-skin (STS) holding on lactation, we compared 24-hour milk volumes of mothers of ventilated low birth weight infants in an STS group to mothers in a non-STS control group. STUDY DESIGN: Mean 24-hour milk volumes at 2, 3, and 4 weeks after delivery of mothers participating in STS holding were compared with those of a retrospective control group from the 12-month period immediately preceding the introduction of STS holding in the neonatal intensive care unit. A repeated-measures analysis of variance adjusting for baseline volumes (1 week after delivery) was used to evaluate the difference in milk volumes between STS and control groups. RESULTS: Sixteen mothers initiated STS holding during the 2-month study period. Eight mothers met study criteria by initiating STS holding during the first 4 weeks after delivery. During a 2-week period the study group had a strong linear increase in milk volume in contrast to no indicative change of the control group's milk volume. CONCLUSION: STS holding of low birth weight infants initiated in the early intensive care phase can result in a significant increase in maternal milk volume, thereby overcoming the frequently seen insufficient lactation experienced by these mothers. PMID- 9210078 TI - Advantages of dual-lumen umbilical vessel catheters versus single-lumen umbilical vessel catheters and additional peripheral intravenous catheters. AB - Once the decision to place an umbilical venous catheter has been made, the opportunity to provide some relief to the patient clinically as well as fiscally is possible with the use of a dual-lumen umbilical vessel catheter (UVC). Although the necessity of catheterization of the umbilical vein remains controversial, successful completion of this procedure provides the practitioner immediate access to the venous circulation of a newborn and allows administration of various solutions and blood products. The cost of a dual-lumen UVC was compared with a single-lumen UVC used in conjunction with peripheral intravenous catheters. By limiting the number of peripheral intravenous catheterization attempts, these patients are spared the pain and possible sequelae associated with wide fluctuations in blood pressure, heart rate, and oxygenation. Once a patient requires more than two peripheral intravenous catheterization attempts, the cost savings of a dual-lumen UVC becomes evident and is amplified with each successive peripheral catheterization attempt. PMID- 9210080 TI - Renal function in full-term neonates with hyperbilirubinemia. AB - The purpose of this study was to investigate the effect of unconjugated hyperbilirubinemia on endogenous creatinine clearance and urinary excretion of sodium, phosphorus, lysozyme, and amino acids in full-term infants. Thirty-seven healthy, breast-fed newborns who were not exposed to phototherapy were studied on their third to fifth day of life. Twenty had neonatal hyperbilirubinemia with a mean indirect bilirubin value of 16.4 mg/dl, whereas 17 who were used as controls had a mean indirect bilirubin value of 7.8 mg/dl. Urine was collected, and samples were taken for examination of creatinine, lysozyme, sodium, and phosphorus concentration. Urinary sediment, glucose, and amino acid levels were also measured. Serum total and direct bilirubin, creatinine, sodium, and phosphorus measurements were taken at the beginning of urine collection. Calculations were made for creatinine clearance, fractional excretion of sodium (FENa), and tubular reabsorption of renal phosphate per deciliter glomerular filtrate (TP/GFR). The means (+/-1 SD) of creatinine clearance, FENa, and TP/GFR were 27.0 +/- 14.2 ml/min/1.73 m2, 0.53% +/- 0.49%, and 5.72 +/- 1.16 mg/dl GF, respectively, in the hyperbilirubinemic group compared with 21.1 +/- 9.4 ml/min/1.73 m2, 0.4% +/- 0.47%, and 6.01 +/- 0.51 mg/dl GF, respectively, in the controls. No statistically significant differences were found between the groups for any of the examined parameters of either glomerular or tubular function. Neonatal hyperbilirubinemia < 20.8 mg/dl has no detrimental effect on renal function of healthy, breast-fed, full-term newborns, and no modification in the approach regarding renal function is necessary in these babies. PMID- 9210079 TI - Intrapartum amniotic fluid index and two-diameter pocket are poor predictors of adverse neonatal outcome. AB - OBJECTIVE: The objective of this study was to determine whether amniotic fluid index < or = 5.0 cm or two-diameter pocket volume < or = 15 cm2 is a predictor of abdominal delivery because of fetal distress or of Apgar score < 7 at 1 or 5 minutes. STUDY DESIGN: The study was prospective and involved 209 parturients in early labor who had ultrasonographic assessment of amniotic fluid volume by both methods. RESULTS: The incidences of cesarean delivery because of fetal distress and of Apgar scores < 7 at 1 and 5 minutes were 8.1%, 9.0%, and 1.9%, respectively. Only Apgar scores < 7 at 1 minute were significantly higher among patients with a two-diameter pocket < 15 cm2 (16/114) as compared with > 15(2) (3/95, p = 0.007). Oligohydramnios by either method was a poor predictor of adverse outcomes (p values ranging from 0.06 to 0.21). Receiver-operating characteristic curves generated for the amniotic fluid index or two-diameter pocket to predict abdominal delivery because of fetal distress or Apgar scores < 7 at 1 minute indicate that both methods are poor diagnostic tests to predict these complications. CONCLUSION: Intrapartum assessment of amniotic fluid volume, by amniotic fluid index or the two-diameter pocket technique, is a poor predictor of adverse neonatal outcome. PMID- 9210081 TI - National standardization of fetal monitoring terminology and competency validation: a call for interdisciplinary collaboration. AB - Although the literature supports standardization of fetal monitoring terminology and practice, national published guidelines have not recommended or outlined a mechanism for accomplishing the goal of standardization. Topics discussed in this article include the effect of fetal monitoring on perinatal morbidity, terminology and significance of fetal monitoring, and the need for, and process of, competency validation. Some individual hospitals have developed competency validation programs for nurses, but less often provide staff physicians with similar opportunities. Recommendations are outlined that include multidisciplinary development of standardized guidelines that consist of mechanisms for competency validation for all obstetric care providers. PMID- 9210082 TI - Perinatal care in Lithuania. AB - The Preventive Program of Perinatal, Neonatal and Congenital Abnormalities in Lithuania was launched in 1992. That was the beginning of the reorganization of the Soviet maternal and child health care system. The first stage of the Program provided for the years 1992 through 1996 and aimed to create a system of maternal and neonatal care; to create a system of diagnosis and prevention of congenital abnormalities; to collect, process, and analyze maternal and neonatal data (to establish a new database); to evaluate, distribute, and use available resources efficiently; to plan financial and human resources for a perinatal care infrastructure; and to train medical personnel and control the level and quality of their knowledge. The reorganization was based on a three-tiered maternal and neonatal care system. By the end of 1996 the major goal of the Program was achieved successfully (i.e., perinatal, neonatal, and infant mortality rates decreased significantly. During the next 5 years the Program will focus mainly on qualitative aspects of perinatal care. PMID- 9210083 TI - Bilateral choanal narrowing as a presentation of craniometaphyseal dysplasia. PMID- 9210084 TI - Successful interferon therapy in a neonate with life-threatening Kasabach-Merritt syndrome. AB - We report a neonatal case of severe, life-threatening Kasabach-Merritt syndrome that was successfully treated with interferon-alpha: This patient had a huge hemangioma of the right leg and a general bleeding tendency. Although the condition initially responded to steroid and radiation therapy, after a relapse, no therapy including steroids, radiation, aspirin, and dipyridamole was effective. Because of severe thrombocytopenia and extension of the hemangioma to the pelvic region, surgical intervention was not indicated. Interferon-alpha therapy was started on day 61 of life. During the therapy the platelet counts increased by more than tenfold and reached the normal level in a month. The size of the hemangioma dramatically decreased. The administration of interferon-alpha might be indicated as a therapy for severe, life-threatening Kasabach-Merritt syndrome, especially when there is resistance to steroid or radiation therapy. PMID- 9210085 TI - Infectious diseases casebook. Pasteurella multocida early onset septicemia in newborns. PMID- 9210086 TI - Special imaging casebook. Walker-Warburg syndrome. PMID- 9210087 TI - Intra-familial transmission and distribution of Prevotella intermedia and Prevotella nigrescens. AB - The periodontal bacteria Prevotella intermedia and Prevotella nigrescens have been recently separated from each other. The purpose of this study was to investigate the distribution and routes of transmission of these bacteria among family members. Seven patients with moderate to severe periodontitis were selected. These probands, their spouses and 14 of their children were investigated. The presence of Pr. intermedia and Pr. nigrescens was determined by culture techniques in pooled subgingival plaque samples, in the saliva, on the tongue, tonsils and buccal mucosa. Differentiation of Pr. intermedia and Pr. nigrescens was performed by enzyme electrophoretic mobility. From all 7 patients, as well as 4 spouses and 3 of the children, Pr. intermedia could be isolated. Pr. nigrescens was found in 2 of the 7 patients, in 5 of the spouses and in 5 of the 6 children aged 5-10 yr. In the 8 children aged 0-4 yr both species were seldom isolated. These data are in accordance with earlier findings that Pr. intermedia is associated with periodontitis and Pr. nigrescens with a relatively healthy periodontal condition. Ribotyping of bacteria was performed by hybridization of HindIII restriction endonuclease digests of chromosomal DNA with ribosomal DNA. Isolates from unrelated individuals always had distinct ribotypes. Indistinguishable ribotypes of Pr. intermedia and Pr. nigrescens were found both among married couples and among parents and children. This indicates that intrafamilial transmission of Pr. intermedia and Pr. nigrescens is possible both between adults and between parents and children. PMID- 9210088 TI - Statistical methods for the estimation of sensitivity and specificity of site specific diagnostic tests. AB - The performance of periodontal diagnostic tests is often evaluated by estimating their sensitivity and specificity with respect to a traditionally accepted standard test regarded as a "gold standard" in making the diagnosis. Correlated samples of binary data arise in dental research. The fundamental unit for analysis is occasionally the site rather than the patient in site-specific dental studies. Statistical methods that take into account the within-patient correlation should be employed to estimate the sensitivity and the specificity of diagnostic tests since site-specific results within a patient can be highly correlated. Several statistical methods are introduced for the estimation of the sensitivity and the specificity of site-specific diagnostic tests; these techniques are applied to the data from a study involving an enzymatic diagnostic test to motivate and illustrate the estimation of the sensitivity and the specificity of periodontal diagnostic tests. PMID- 9210089 TI - Identification of calprotectin, a calcium binding leukocyte protein, in human dental calculus matrix. AB - Calprotectin is a calcium binding protein produced by leukocytes, macrophages and epithelial cells, and its levels in several tissues increase during infections and in many inflamed areas, suggesting that it may be an indicator of inflammatory activity. Osteopontin is a prominent phosphorylated glycoprotein in bone matrix, having calcium binding capacity. Recently, it has been reported that calprotectin and osteopontin are present in urinary stones (pathological mineralized masses in the body), and that these proteins may be involved in their formation. Dental calculus formed by mineralization of dental plaque is an inflammatory factor which may contribute to periodontal disease. It contains many organic components involved in mineralization. We recently found osteopontin molecules in human dental calculus and suggested that the components of its matrix may be similar to those of urinary stones. In this study, we investigated the presence of calprotectin in human dental calculus by immunohistochemical and immunoblotting analyses using a specific antibody for calprotectin. After fixation and demineralization of dental calculi adhered to tooth roots, sections embedded in paraffin were immunoreacted with the antibody for calprotectin and positive immunostaining for calprotectin was observed. Dental calculus proteins were then extracted with EDTA and separated by electrophoresis on 15% polyacrylamide gels. By immunoblotting analysis, 3 or 4 bands were observed at 11, 14.5, 22-25, 28 or 36.5 kDa and these patterns corresponded to those of calprotectin subunits. When non-immune rabbit serum was used instead of calprotectin-specific antibody as a negative control, no immunoreactivity was observed. These findings indicate that calprotectin is associated not only with antibacterial action but also with calcium binding capacity during dental calculus formation. PMID- 9210090 TI - The modulatory role of cementum matrix on osteoblastic cells in vitro. AB - The formation of new cementum is an important issue in clinical periodontology, as cementum is required to provide attachment for newly formed periodontal tissues to the root surface. In this study a model of cementogenesis in vitro was used in order to test the effects of root surface demineralization on the migration, attachment and formation of a cementum-like tissue by osteoblastic cells cultured on cementum and to test the specificity of cementum matrix in modulating those effects by comparison of root co-cultures with bone co-cultures. It was demonstrated that root surface demineralization did not significantly alter the orientation, number and attachment of cells to the root co-cultures. The results also demonstrated that cementum and bone matrix appear to behave differently in culture, as seen by their distinct action on the morphological profile of the attached cells and the extracellular matrix deposited by these cells. These results demonstrate that although cementum matrix appears to stimulate the production of cementum-like tissue, this action is not confined to cementum matrix alone, since a similar material was also deposited on dentine and bone surfaces. Thus, these results do not support a specific action of cementum matrix on the modulation of the cementoblast phenotype. The use of co-cultures of neonatal rat calvaria cells with root slices represents a promising model of cementogenesis in vitro; however, studies should be undertaken towards the identification of markers to distinguish between cementoblast and osteoblast phenotypes in order to further validate this model. PMID- 9210091 TI - Absence of adult dental anomalies in familial hypophosphatasia. AB - This paper is a supplemental report on 3 previous publications about a family in which 3 male children manifested gingival recession, alveolar bone resorption and premature exfoliation of their deciduous teeth without apical root resorption and without clinical signs of inflammation. Laboratory blood and urine studies in conjunction with an analysis of periodontal microflora and family pedigrees established a diagnosis of hypophosphatasia in these 3 children, as well as their father, the paternal grandmother and paternal great-uncle. Clinical data also revealed that a son of the paternal great-uncle and his daughter were similarly affected. The family pedigree is consistent with an autosomal dominant mode of transmission. The 3 brothers are now between the ages of 18 and 22 yr and all have complete permanent dentitions. Aside from some periodontal manifestations of prior dentoalveolar trauma, most of the findings of the periodontal assessment are within normal limits. All 3 exhibit moderate to severe caries and some degree of gingival inflammation, but minimal periodontal pathosis. PMID- 9210092 TI - The effect of race, smoking and immunoglobulin allotypes on IgG subclass concentrations. AB - In previous studies we have demonstrated that serum IgG subclass concentrations are influenced by both race and periodontal disease diagnosis. Furthermore, we have shown that smoking habits modify the concentrations of some IgG subclasses in specific racial and diagnostic groups. In view of a large amount of data showing strong associations between immunoglobulin allotypes and IgG subclass concentrations we have investigated the effects of race, smoking and IgG allotype on IgG subclass concentration in a population of subjects with or without various forms of periodontitis. The results indicated that there are complex relationships between these factors in their effects on individual IgG subclass levels, and that effects unique to black or white subject groups, or to specific periodontal diagnostic groups and racial subgroups, were evident. In blacks with chronic adult periodontitis IgG1 was lower in smokers, while in generalized early onset periodontitis patients IgG2 was lower in smokers. IgG4 was independently affected by gender (males higher), smoking (smokers lower) and GM23 (GM23 positive subjects higher), in black subjects only. In white subjects, complex relationships between smoking and allotypic markers were noted but no influence of periodontal diagnosis was found. White GM23 negative subjects who smoked had lower levels of IgG1 than GM23 positive subjects. White GM2 negative subjects who smoked had lower levels of IgG2, than did those who did not smoke. In contrast, smoking had no effect on IgG2 levels in GM2 positive subjects. Thus, in addition to immunoglobulin allotype, smoking is associated with IgG subclass concentrations; furthermore, in black subjects, periodontal diagnosis, gender and smoking all influence IgG subclass concentrations. These results demonstrate that genetic and environmental factors can interact to influence levels of individual subclasses. PMID- 9210093 TI - Histomorphological and biochemical differentiation capacity in organotypic co cultures of primary gingival cells. AB - To establish a three-dimensional in vitro test system mimicking the physiological situation of the oral cavity, organotypic co-cultures consisting of primary gingival cells on a collagen matrix with fibroblasts were generated. The histomorphological development after 7 and 14 d revealed close similarity with the non-keratinized gingiva epithelium. Furthermore, as epithelial specific markers synthesis and localization of keratins as well as the deposition of basement membrane components were assessed on frozen sections by immunofluorescence and keratin expression by in situ hybridization. Primary keratinocytes in conventional culture strained positive for keratin K14 and the mucosal differentiation-specific keratins K4 and K13, while primary fibroblasts, isolated from the same tissue source, and also some keratinocytes, were positive for vimentin. In organotypic co-cultures the keratinocytes formed a multilayered epithelium within 14 d containing basal cells and flattened cells in the uppermost layers. Comparable to native non-keratinized gingiva keratin 14 gene expression was clearly detectable in the basal cell compartment but showed extending immunolocalization. In addition, particularly at the early stage (7 d), basally located keratinocytes were also vimentin positive. According to morphological differentiation K4 and K13 were detectable in suprabasal position a the RNA and protein level. The major basement membrane constituents collagen type IV and laminin increased with time revealing first an interrupted and later a fully extended staining underneath the basal cells. Maintenance of basal cell function was further demonstrated by cell proliferation (BrdU incorporation) which was initially high (7 d) but declined towards the later stages (14-21 d). The results demonstrate i) that this co-culture system leads to a stratified surface epithelium with morphological and biochemical characteristics of the non keratinized gingiva epithelium and ii) that a state of physiological tissue balance was reached, thus rendering a suitable model for tissue compatibility studies. PMID- 9210094 TI - Evidence of an association between functional abnormalities and defective diacylglycerol kinase activity in peripheral blood neutrophils from patients with localized juvenile periodontitis. AB - Peripheral neutrophils from patients with localized juvenile periodontitis (LJP) show functional abnormalities, such as impaired locomotion and enhanced respiratory burst activity. A defect in intracellular signalling mechanism has been proposed to be responsible for some changes, but direct evidence is lacking. In this study we have determined the activity of diacylglycerol (DAG) kinase, an enzyme controlling the DAG/protein kinase C (PKC) pathway, in crude cytosolic and membrane fractions of neutrophils from 5L JP patients and age and gender-matched normal individuals. No difference was observed in the DAG kinase activity in subcellular fractions from unstimulated cells between the 2 groups. When normal neutrophils were stimulated with N formyl-methionyl-leucyl-phenylalanine (FMLP), the enzyme activity was markedly increased in both subcellular fractions. In contrast, neutrophils from 3 of the 5 LJP patients tested completely failed to rise the DAG kinase activity upon chemoattractant stimulation. These data indicate that in some LJP patients the neutrophil DAG kinase may be defective. To examine whether a decrease in DAG kinase activity could account for some neutrophil abnormalities seen in LJP, normal neutrophils were treated with R59022, a DAG kinase inhibitor, that has been shown to reduce DAG kinase activity in human neutrophils. Upon stimulation with FMLP, R59022-treated normal neutrophils showed significantly reduced chemotactic response and enhanced respiratory burst activity, two typical functional abnormalities featured by LJP cells. It is concluded that a defect in DAG kinase may cause, through an abnormal accumulation of the endogenous PKC activator DAG some of the functional changes observed in neutrophils from LJP patients. PMID- 9210095 TI - Odor discrimination of 'cAMP-' and 'IP3-increasing' odorants at high temperature and at high NaCl concentration in turtle olfactory system. AB - The ability of the turtle olfactory system to discriminate between various cAMP- and IP3-increasing odorants at high temperature and at high NaCl concentration in the olfactory bulb was examined by the cross-adaptation technique. The degrees of discrimination in high [Na+] solution were similar to those in normal Ringer's solution, suggesting that selectivities of receptors coupled with cAMP- and IP3 dependent pathways are similar to those coupled with both cAMP- and IP3 independent pathways. The mean values of the degree of discrimination among the IP3-increasing odorants were higher than those among the cAMP-increasing odorants at high temperature and at high [Na+] concentration. The degrees of discrimination among the IP3-increasing odorants at 40 degrees C were greater than those at 25 degrees C, while those among the cAMP-increasing odorants at 40 degrees C were similar to those at 25 degrees C, suggesting that the features of the receptors of cAMP-increasing odorants are different from those which respond to IP3-increasing odorants. PMID- 9210096 TI - The indomethacin-induced gastric mucosal damage in rats. Effect of gastric acid, acid inhibition, capsaicin-type agents and prostacyclin. AB - In pylorus-ligated rats subcutaneous (s.c.) pentagastrin (325.5 nmol/kg) or histamine (54.3 mumol/kg), but not the cholinergic agent bethanechol (7.6 or 15.2 mumol/kg), increased gastric mucosal injury by s.c. indomethacin (55.8 mumol/kg). Intragastric (i.g.) administration of 0.15 or 0.3 N HCl greatly potentiated injury by s.c. indomethacin with widespread ulceration, intragastric bleeding and even perforation. The gastric mucosal damage produced by indomethacin plus 0.3 N HCl was reduced by i.g. capsaicin (3.1-25.1 microM), i.g. resiniferatoxin (0.38 6.1 microM), by s.c. atropine (0.15-1.2 mumol/kg) and to a lesser extent by i.g. prostacyclin (40-267 microM) or s.c. cimetidine (198.2 mumol/kg). The protective effect of capsaicin or resiniferatoxin was not prevented by atropine or cimetidine treatment. Capsaicin (6.5 mM) enhanced gastric injury by s.c. or i.g. indomethacin. Results indicate the importance of early vascular events in the pathogenesis of mucosal injury induced by indomethacin in the stomach and suggest a role for gastric acid in potentiation of such injury. Results further strengthen the idea of a protective role for capsaicin-sensitive sensory nerves in the stomach. PMID- 9210099 TI - Comparative study of .OH generation in brain, heart and liver using microdialysis. PMID- 9210098 TI - Ibotenate lesion of the medial hypothalamus alters the salt intake and pressor responses to activation of the median preoptic nucleus in rats. AB - We investigated the influence of ibotenic acid lesions of the medial hypothalamus (MH) on salt appetite and arterial blood pressure responses induced by angiotensinergic and adrenergic stimulation of the median preoptic nucleus (MnPO) of rats. Previous injection of the adrenergic agonists norepinephrine, clonidine, phenylephrine, and isoproterenol into the MnPO of sham MH-lesioned rats caused no change in the sodium intake induced by ANG II. ANG II injected into the MnPO of MH-lesioned rats increased sodium intake compared with sham-lesioned rats. Previous injection of clonidine and isoproterenol increased, whereas phenylephrine abolished the salt intake induced by ANG II into the MnPO of MH lesioned rats. Previous injection of norepinephrine and clonidine into the MnPO of sham MH-lesioned rats caused no change in the mean arterial pressure (MAP) induced by ANG II. Under the same conditions, previous injection of phenylephrine increased, whereas isoproterenol reversed the increase in MAP induced by angiotensin II (ANG II). ANG II injected into the MnPO of MH-lesioned rats induce a decrease in MAP compared with sham-lesioned rats. Previous injection of phenylephrine or norepinephrine into the MnPO of MH-lesioned rats induced a negative MAP, whereas pretreatment with clonidine or isoproterenol increased the MAP produced by ANG II injected into the MnPO of sham- or MH-lesioned rats. These data show that ibotenic acid lesion of the MH increases the sodium intake and pressor responses induced by the concomitant angiotensinergic, alpha 2 and beta adrenergic activation of the MnPO, whereas alpha 1 activation may have opposite effects. MH involvement in excitatory and inhibitory mechanisms related to sodium intake and MAP control is suggested. PMID- 9210097 TI - Convulsive effects of a benzodiazepine receptor inverse agonist: are they related to anxiogenic processes? AB - The linkage-testing strain of ABP/Le mice carries six mutations which express in easily identifiable phenotypes. By crossing this strain with a traditional inbred strain (C57BL/6ByJ) which is the 'wild type' for the mutated ABP/Le loci, we produced Mendelian populations, intercrosses and backcrosses so as to estimate whether the sensitivity to methyl beta-carboline-3-carboxylate (beta-CCM), a benzodiazepine receptor inverse agonist, and anxiety-related behaviour could be related to a common genetically determined substrate. We have shown that one locus on chromosome 9 is associated with beta-CCM-induced seizures and three loci on chromosomes 4, 7 and 9 are associated with anxiogenic processes. Analysis of [3H]flumazenil binding suggested a possible involvement of a Bmax decrease in both beta-CCM-induced seizures and anxiogenic processes. The putative common genetic regulation of both mechanisms is discussed. PMID- 9210100 TI - Considering a decision-making approach to youth violence prevention programs. AB - The unfortunate exception to a general downturn in violent crime involves an upsurge in violence among youth. Violence often results when minor confrontations escalate. As school violence increasingly has become widespread, schools have become the location of many violence prevention efforts, few of which have been evaluated adequately. This paper focuses on enhancing decision-making skills as one approach to increase adolescents' ability to manage interpersonal violence. Adolescents can be considered fairly skilled decision-makers and their unique perspective must be considered in development of effective intervention programs. Data from a pilot study were examined for insights about adolescents' ability to make decisions in situations of interpersonal conflict. PMID- 9210101 TI - Nurses' logs as an evaluation tool for school-based violence prevention programs. AB - Programs for preventing violence among youth should be evaluated to determine if they are effective. Nurses' logs appear to be a useful tool for evaluating school based violence prevention programs. The logs provide a record of students' visits to the school nurse that can be used to determine if a violence prevention program is associated with a reduction in fighting- and other injury-related nurse visits. This method has many strengths: it is simple and inexpensive, it does not interrupt the school routine, it permits school-level rather than student-level data collection, it provides a ready "baseline," and it allows continuous data collection. However, potential limitations do exist. For example, the method may provide insufficient information and may be affected by factors unrelated to the intervention. School officials can increase the usefulness of the logs by encouraging standardization and providing training in their use. PMID- 9210102 TI - AIDS among children--United States, 1996. Division of HIV/AIDS Prevention, CDC. PMID- 9210103 TI - Perceived risk of fighting and actual fighting behavior among middle school students. AB - This study evaluated the association between perceived risk of fighting and actual fighting behavior among middle school students and determined if that relationship was modified by race, gender, or grade level. Survey data were obtained from a stratified random sample of 517 Black and White students in a county school district in Maryland. Most students (72%) perceived fighting to be high-risk, but 20% reported fighting on a regular basis. The effect of risk perception (RP) on fighting behavior varied by race. Independent of grade and gender effects, students who believed fighting to be low-risk were more likely to fight on a regular basis than those with high RP (odds ratio for Blacks = 3.1; odds ratio for Whites = 5.4). School violence prevention education must include an emphasis on the health risks of fighting and attention to cultural differences in risk perception. PMID- 9210104 TI - Latino families: partners for success in school settings. PMID- 9210105 TI - Working with education reporters to advocate for comprehensive school health education. PMID- 9210106 TI - The needs of children and the role of school nurses. PMID- 9210107 TI - Symbolic play of children with language impairment: a critical review. AB - There have been a number of studies that have reported on the symbolic play abilities of children assessed as demonstrating developmental language disorders or specific language impairment. In general, this research has reported significant differences in the symbolic play abilities of children with language impairment and those developing language normally. In most, though interestingly, not all cases, the differences reflected less developed symbolic play of the children with language impairments. It will be argued here that these reported differences should not be interpreted as demonstrative of marked deficits in the general representational or specific symbolic play competence of children with language impairments. It will be argued further that part of the research conducted to date on the symbolic play abilities of children with language impairment has been confounded by the encroachment of language into the research procedures, that the level of play often investigated has not been unquestionably symbolic in nature, and that the actual differences in symbolic play have not been substantial. PMID- 9210108 TI - An examination of the morpheme BE in children with specific language impairment: the role of contractibility and grammatical form class. AB - This study examined the production of the morpheme BE, focusing on the influence of contractibility, the relationship between copula and auxiliary forms, and the occurrence of non-omission errors. Language samples collected from children with SU and from normal language learners at equivalent MLU levels were analyzed. Three levels of contractibility were examined: contractible, syntactically uncontractible, and phonetically uncontractible. Contractible contexts were produced significantly more accurately than uncontractible contexts by both groups. There was no difference between the two forms of uncontractibility. Furthermore, there were no significant interactions between language status and contractibility, suggesting that contractibility influenced both groups equally. Copula forms were produced more consistently than auxiliary. There was no interaction between BE type and language status. The groups did not differ in proportion or type of non-omission error. The results are discussed in relation to accounts of morphological deficits in SU. PMID- 9210109 TI - The efficacy of computer-provided reading treatment for chronic aphasic adults. AB - We examined the effects of computer-provided reading activities on language performance in chronic aphasic patients. Fifty-five aphasic adults were assigned randomly to one of three conditions: computer reading treatment, computer stimulation, or no treatment. Subjects in the computer groups used computer 3 hours each week for 26 weeks. Computer reading treatment software consisted of visual matching and reading comprehension tasks. Computer stimulation software consisted of nonverbal games and cognitive rehabilitation tasks. Language measures were administered to all subjects at entry and after 3 and 6 months. Significant improvement over the 26 weeks occurred on five language measures for the computer reading treatment group, on one language measure for the computer stimulation group, and on none of the language measures for the no-treatment group. The computer reading treatment group displayed significantly more improvement on the Porch Index of Communicative Ability "Overall" and "Verbal" modality percentiles and on the Western Aphasia Battery Aphasia "Quotient" and "Repetition" subtest than the other two groups. The results suggest that (a) computerized reading treatment can be administered with minimal assistance from a clinician, (b) improvement on the computerized reading treatment tasks generalized to non-computer language performance, (c) improvement resulted from the language content of the software and not stimulation provided by a computer, and (d) the computerized reading treatment we provided to chronic aphasic patients was efficacious. PMID- 9210110 TI - A preliminary account of the effect of otitis media on 15-month-olds' categorization and some implications for early language learning. AB - In the present study, 24 infants 14 to 16 months old (M = 15.3 months) with a history of otitis media (OM) and tube placement were tested for categorical responding within a visual familiarization-discrimination paradigm (cf. Roberts & Jacob, 1991; Roberts, 1995a). Sixteen infants were, on average, 4.6 months post tubes. At the time of testing, hearing was reported to be normal. During both the familiarization and testing phases, reiterant speech ulterances (e.g., "ti ti the sasasa," "bo bo the sasasa," "gu gu the sasasa," "sasasa") were randomly presented contingent on infants' looking at line drawings of selected animals. The inclusion of the article the in these utterances provides an important grammatical cue signaling an upcoming noun label. Using these same procedures, utterances, and animal stimuli, normal-hearing 15-month-olds without a reported history of OM had categorized successfully in two previous experiments. In contrast, the OM infants did not respond categorically under these conditions. This suggests that even mild hearing loss may adversely affect categorical responding under specific input conditions and that this effect may persist after normal hearing is restored. A plausible account is that fluctuating hearing loss may cause the low-phonetic-substance article to vacillate above and below some attentional threshold. The resulting inconsistency in the article's availability would disrupt the co-occurrence relation between the article and cues already connected to joint attentional/referential interactions and categorical treatment of objects (cf. Roberts, 1995a). Such a disruption has implications for the acquisition of words and other aspects of language. PMID- 9210111 TI - Reducing bias in language assessment: processing-dependent measures. AB - One potential solution to the problem of eliminating bias in language assessment is to identify valid measures that are not affected by subjects' prior knowledge or experience. In this study, 156 randomly selected school-age boys (31% majority; 69% minority) participated in three "processing-dependent" language measures, designed to minimize the contributions of prior knowledge on performance; and one traditional "knowledge-dependent" language test. As expected, minority subjects obtained significantly lower scores than majority participants on the knowledge-dependent test, but the groups did not differ on any of the processing- dependent measures. These results suggest that processing dependent measures hold considerable promise for distinguishing between children with language disorders, whose poor language performance reflects fundamental psycholinguistic deficits, and children with language differences attributable to differing experiential backgrounds. PMID- 9210112 TI - Truncation patterns in English-speaking children's word productions. AB - This study examines English-speaking children's truncation patterns (i.e., syllable deletion patterns) in multisyllabic words to determine if they are consistent with metrical constraints or perceptual biases. It also examines segmental influences on children's truncations. Children, age 22-34 months, produced three-syllable novel and real words and four-syllable real words, which varied across stress and segmental pattern. Results revealed a significant stress pattern effect on truncation rate, but findings were not consistent with metrical or perceptual salience predictions. The clearest account of the findings came from an analysis of truncation rate across individual words: Children truncated WSW (weak-strong-weak) words and words that contained intervocalic sonorants more frequently than other words. Analysis of truncation patterns in SWW and SWSW words revealed that final unstressed syllables were more frequently preserved than nonfinal unstressed syllables. Findings support the interaction between metrical, syllabic, and acoustic salience factors in children's multisyllabic word productions. PMID- 9210113 TI - Determinants of sentence comprehension in aphasic patients in sentence-picture matching tasks. AB - The results of two studies of sentence comprehension in aphasic patients using sentence-picture matching tests are presented. In the first study, 52 aphasic patients were tested on 10 sentence types. Analysis of the number of correct responses per sentence type showed effects of syntactic complexity and number of propositions. Factor analysis yielded first factors that accounted for two-thirds of the variance in performance to which all sentence types contributed. Clustering analysis yielded groups of patients whose performances progressively deteriorated and in which performance was more affected by sentence types that were harder for the group overall. These results were very similar to those previously obtained using an enactment task. In the second study, 17 aphasic patients were tested on the same 10 sentence types using both sentence-picture matching and enactment tasks. Correlational analyses showed that performance on the two tests was significantly correlated across both subjects and sentences. The results provide data relevant to the determinants of the complexity of a sentence in auditory comprehension. PMID- 9210114 TI - Late talkers at 2: outcome at age 3. AB - Age 3 follow-up data are presented for a sample of 34 toddlers diagnosed between the ages of 24 and 31 months with expressive type specific language impairment (SU-E). At age 3, the late talkers scored significantly lower on all language measures than 21 comparison peers matched at intake on age, SES, and nonverbal ability. When seen at follow-up, the former late talkers scored in the average range on the Expressive One-Word Picture Vocabulary Test (EOWPVT) and on the Reynell Expressive Language Scale, but more than 1.5 SDs below age expectations in MLU and on Scarborough's (1990a) IPSyn. The proportion of late talkers performing in the average range at follow-up varied markedly as a function of measure used (EOWPVT: 79%, Reynell: 58%, MLU: 35%, and IPSyn: 24%), indicating that the late talkers made more rapid progress in lexical development and in the use of language to define, explain, and describe than they did in the areas of syntactic and morphological development. The only significant predictor of age 3 outcome was intake expressive language level, with toddlers who had been more severely delayed in expressive language at intake relative to age level having the worst outcomes at age 3. PMID- 9210115 TI - The genetic basis of persistence and recovery in stuttering. AB - Although past research has provided evidence of a genetic component to the transmission of susceptibility to stuttering, the relationship between the genetic component to stuttering and persistence and recovery in the disorder has remained unclear. In an attempt to characterize this relationship, the immediate and extended families of 66 stuttering children were investigated to determine frequencies of cases of persistent and recovered stuttering. Pedigree analysis and segregation analysis were used to examine patterns of transmission. The following questions were investigated: 1. Is there a sex effect in recovery from stuttering? Here, we sought to test the hypothesis that females are more likely to recover than males, leading to the change in sex ratio from approximately 2:1 males to females close to onset of the disorder, to 4 or 5:1 in adulthood. 2. Is persistence/recovery in stuttering transmitted in families? If recovery/ persistence appears to be transmitted, (a) are recovered and persistent stuttering independent disorders?; (b) is recovery a genetically milder form of persistent stuttering?; or (c) is persistence/recovery transmitted independent of the primary susceptibility to stuttering? Results indicated sharply different sex ratios of persistent versus recovered stutterers in that recovery among females is more frequent than among males. It was found that recovery or persistence is indeed transmitted, and further, that recovery does not appear to be a genetically milder form of stuttering, nor do the two types of stuttering appear to be genetically independent disorders. Data are most consistent with the hypothesis that persistent and recovered stuttering possess a common genetic etiology, and that persistence is, in part, due to additional genetic factors. Segregation analyses supported these conclusions and provided statistical evidence for both a single major locus and polygenic component for persistent and recovered stuttering. PMID- 9210116 TI - Identifying the authoritative judgments of stuttering: comparisons of self judgments and observer judgments. AB - Reliable and accurate stuttering measurement depends on the existence of unambiguous descriptions or exemplars of stuttered and nonstuttered speech. The development of clinically meaningful and useful exemplars, in turn, requires determining whether persons who stutter judge the same speech to be stuttered that other observers judge to be stuttered. The purpose of these experiments, therefore, was to compare stuttering judgements from several sources: 15 adults who stutter, judging their own spontaneous speech; the same adults who stutter, judging each other's speech; and a panel of 10 authorities on stuttering research and treatment. Judgments were mode under several conditions, including self judgments made while the speaker was talking and self- and other-judgements made from recordings in continuous and interval formats. Results showed substantial differences in stuttering judgments across speakers, judges, and judgment conditions, but across-task comparisons were complicated by low self-agreement for many judges. Some intervals were judged consistently by all judges to be Stuttered or Nonstuttered, across multiple conditions, but many other intervals were either not assigned replicable judgments or were consistently judged to be Nonstuttered by the speaker who had produced them but were not assigned consistent judgments by other judges. The implications of these findings for stuttering measurement are considered. PMID- 9210117 TI - Developmental changes in laryngeal and respiratory function with variations in sound pressure level. AB - The development of the speech production system was investigated using a cross sectional design that included children aged 4-14 years and adults. Given that the size and internal structure of the laryngeal and respiratory systems differ between children and adults, it was predicted that children would show functional distinctions from adults during speech. Aerodynamic, acoustic, and respiratory kinematic techniques were used to assess laryngeal and respiratory function while participants varied their sound pressure level. In general, the aerodynamic and acoustic results show that men and 14-year-old boys function differently than women and all other groups of children. For the respiratory function data, children's values are similar to adults' by the time they are 12-14 years of age. These changes correspond closely to developmental laryngeal and respiratory anatomic data. All participants used a combination of laryngeal and respiratory mechanisms to increase sound pressure level, but the combination of mechanisms differed across age groups. These data emphasize that the laryngeal and respiratory behavior of children is not easily predicted from an adult model. PMID- 9210118 TI - Preliminary voice and speech analysis following fetal dopamine transplants in 5 individuals with Parkinson disease. AB - A surgical procedure involving transplantation of fetal dopamine cells into the striatum of persons with advanced Parkinson disease (PD) has recently been performed in an attempt to alleviate Parkinsonian and drug-dose related symptoms (e.g., the "on-off" phenomena). Improvements in limb motor and neurological function, as well as less severe and shorter on-off episodes have been reported following fetal cell transplant (FCT) surgery. Acoustic, electroglottographic, and perceptual measures were analyzed pre- and post-surgery to determine if phonotory and articulatory function were affected by this relatively new form of treatment. In addition, speech and motor exam measures were compared to determine if similar directional changes across motor systems were apparent. Findings suggest that FCT surgery did not systematically influence voice and speech production. Also, it appears that FCT surgery may differentially affect phonatory, articulatory, and limb motor systems. Findings are discussed relative to these differential effects. PMID- 9210119 TI - Disfluency in spasmodic dysphonia: a multivariate analysis. AB - This study examined visual analog scaling (VAS) judgments of disfluency by normal listeners in response to oral reading by speakers with spasmodic dysphonia (SD) and by nondysphonic controls, as well as the variables of frequency of occurrence of disfluencies, speaking rate, number of reading errors, and temporal acoustic measures of interword interval duration and articulation time. MANOVA yielded statistically significant differences between SD and control speakers for all variables except reading errors. Although no significant fluency-related differences were observed in terms of type of vocal spasm or voice tremor, significant differences in disfluency measures were obtained for clinical ratings of severity of dysphonia. Greater dysphonia severity ratings were associated with decreased fluency, but milder ratings were not necessarily associated with disfluency. Stepwise multiple regression analysis demonstrated that frequency of disfluency occurrence, speaking rate, and reading errors accounted for more than three fourths of the variability in VAS judgments of disfluency. Findings suggest that although disfluency is not a defining feature of SD, it does contribute significantly to the overall clinical impression of severity of the disorder. PMID- 9210120 TI - A preliminary investigation of the effects of gender and race on Voice Onset Time. AB - Twenty individuals participated in a study of Voice Onset Time (VOT) production. Participants included equal numbers of males and females and equal numbers of African Americans and Caucasian Americans. Each individual read a set of stimuli formed from the six stop consonants (/p/,/t/,/k/;/b/,/d/,/g/) combined with the three vowels /i/,/a/, and /u/. Their productions were measured for VOT. Considerably more prevoicing (i.e., negative VOT) for voiced stops was found in the present study in comparison with past studies. Statistically significant differences were found for both gender and race. These results suggest that the normative data presently available is probably inadequate because it does not accurately reflect the normal distribution of either gender or race within the American population. PMID- 9210121 TI - Acoustic cues to place of articulation and the McGurk effect: the role of release bursts, aspiration, and formant transitions. AB - The McGurk effect demonstrates that the perceived place of articulation of an auditory consonant (such as/bi/) can be influenced by the simultaneous presentation of a videotape of a talker saying a conflicting consonant such as /gi/. Usually, such a presentation is perceived by observers as "di" or "delta i" (known as fusion responses). The reverse pairing (auditory /gi/ paired with a visual /bi/) results in "bgi." percepts. These are known as combination responses. In the current study, three experiments examined how acoustic information about place of articulation contained within the release bursts, aspiration, and voiced formants and transitions of a consonant contribute to the McGurk effect. In the first experiment, the release bursts and aspiration were deleted from the acoustic signal. This manipulation resulted in a smaller impact on McGurk "fusion" tokens relative to the McGurk "combination" tokens. This asymmetry may be related to the perceptual salience of the release bursts and aspiration for velar compared to the bilabial tokens used in this experiment and their importance for obtaining the combination percept. In Experiment 2, the release bursts and aspiration were increased in amplitude. Results revealed either no effect or a stronger McGurk effect for the manipulated tokens than for the intact tokens. This findings suggests that the McGurk effect for fusion tokens does not occur simply because the release bursts and aspiration are weak. In Experiment 3, low-pass filtering the second and higher formants and transitions was associated with the largest overall impact on the McGurk effect. This suggests that dynamic information contained within these formants is of primary importance in obtaining the McGurk effect. These cues are, however, context-dependent and vary as a function of talker and vowel context. PMID- 9210122 TI - A comparison of the benefit provided by well-fit linear hearing aids and instruments with automatic reductions of low-frequency gain. AB - In this clinical study, 110 patients seen at three different clinical facilities were fit binaurally with linear, in-the-canal (ITC) hearing aids. All patients were new hearing aid users. Each of the hearing aids was equipped with an adjustable control that could be set by one of the audiologists (Audiologist A) at each site to convert it from a linear instrument to an experimental nonlinear one with automatic reduction of low-frquency gain at high input levels (or base increase at low levels, BILL). Both the patient and the audiologist performing the outcome testing at each site (Audiologist B) were blind as to the present setting of the hearing aid. Each participant was enrolled in the study for a total of 12 weeks, with the hearing aid set to either the linear or BILL processing mode of operation for the first 8 weeks and the opposite setting for a subsequent 4-week period. In summary, this was a prospective, doubleblind, crossover study of 110 new hearing aid users. Outcome measures focused on hearing aid benefit and included both objective and subjective measures. Objective measures were derived from scores on the Northwestern University Auditory Test NO. 6 (NU-6) and the Connected Speech Test (CST) obtained for all possible combinations of two speech presentation levels (60 and 75 dB SPL) two types of background noise (cafeteria noise and multitalker babble), and two signal-to noise ratios (+5 and +10 dB). Subjective outcome measures included magnitude estimation of listening effort (MELE), the abbreviated form of the Hearing Aid Performance Inventory (HAPI), and estimations of hearing-aid usage based on daily use logs kept by the participants. All of these measures were used to evaluate the benefit provided by linear amplification and the benefit resulting from the experimental BILL processing. Participant preferences for the experimental BILL processing scheme or linear processing were also examined by using a paired comparison task at the end of the study. Results were analyzed separately for three subgroups of patients (mild, moderate, severe) formed on the basis of their average hearing loss at 500, 1000, 2000, and 4000 Hz. In all three subgroups, significant improvement in performance was observed for linear amplification and for BILL processing when compared to unaided performance. There were no significant differences in aided performance, however, between linear processing and the experimental BILL processing. PMID- 9210123 TI - Age of second-language acquisition and perception of speech in noise. AB - To determine how age of acquisition influences perception of second-language speech, the Speech Perception in Noise (SPIN) test was administered to native Mexican-Spanish-speaking listeners who learned fluent English before age 6 (early bilinguals) or after age 14 (late bilinguals) and monolingual American-English speakers (monolinguals). Results show that the levels of noise at which the speech was intelligible were significantly higher and the benefit from context was significantly greater for monolinguals and early bilinguals than for late bilinguals. These findings indicate that learning a second language at an early age is important for the acquisition of efficient high-level processing of it, at least in the presence of noise. PMID- 9210124 TI - Vocal reaction times of children with CAPD, age-matched peers, and young adults to printed words. AB - The purpose of this study was to determine if there were differences between children identified in a clinical setting as having Central Auditory Processing Disorder (CAPD), an age-matched peer group, and young adults when tested using a vocal reaction time (VRT) format. The children with CAPD were matched by gender and age to peers between the ages of 8 and 10 years. All speakers were presented visually with printed third-grade-level one- and two-syllable words (e.g., boy, mother) as well as the syllable "uh". Participants spoke each word according to the criteria of seven separate conditions, which included immediate naming tasks (0 s delay), a short delay before speaking (M = 1.5 s), and a longer delay before speaking (M = 4.0 s). Speakers VRTs were measured, and production errors were recorded. All speakers took longer to respond in the immediate-response conditions than the delayed-response conditions. Statistically significant differences were found for the immediate-response conditions, with means for the children with CAPD reflecting slower performance than that of their peers. The peer group was slower than the adults. For the delayed conditions, both groups of children responded with significantly longer VRTs than the adults. The two groups of children did not differ for these tasks. The children with CAPD produced a significantly greater number of errors than their peers, specifically for the long-delay conditions. The adults showed no performance differences across the immediate response conditions nor across the delayed conditions. These results suggested that children with CAPD may have processing difficulties with visual stimuli. PMID- 9210126 TI - Early operative reconstruction of severe ligamentous knee injuries in patients with multiple trauma. AB - Although operative reconstruction of severe knee ligament injuries has been the preferred method of treatment, the timing of that reconstruction relative to the injury has not been previously addressed. In a retrospective review of multitrauma cases, we identified 32 patients with 34 severe knee ligament injuries. Of these 34 knees, 19 (56%) were treated with early reconstruction (within 2 weeks of injury). Of the remaining 15 knees, 8 were managed by delayed reconstruction at an average of 4.8 months after injury, and 7 knees were not surgically treated. All 19 knees treated by early operation were clinically stable, and 89% were pain free at an average of 25 months' follow-up. In contrast, only 13% of the knees treated by delayed or no reconstruction were clinically stable, 33% were persistently painful, and 40% required bracing intermittently to allow activities of daily living. In this study, patients treated by delayed reconstruction or non-operatively had poor functional results. We believe early operative reconstruction of severe knee injuries in multitrauma patients is crucial to maximize functional outcome and minimize long-term sequelae. PMID- 9210125 TI - Paravertebral inflammation mistaken for neoplasm or abscess by MRI. AB - Magnetic resonance imaging (MRI) is a useful procedure for the evaluation of spinal pathology in children. The high sensitivity of soft tissue changes may lead to misinterpretation of paravertebral swelling caused by inflammation. We present a report of three patients with paravertebral soft tissue swelling caused by disk space infection or intervertebral disk space calcification. The findings shown by MRI were initially interpreted as neoplasm or abscess, which led to surgical procedures either planned or done. Surgeons need to exercise extreme care when interpreting soft tissue changes shown by MRI. PMID- 9210127 TI - Bilateral pseudarthrosis of the olecranon. PMID- 9210128 TI - Avascular necrosis of the femoral head after intramedullary nailing of a femoral shaft fracture in a male adolescent. PMID- 9210129 TI - Tumoral calcinosis in an infant. PMID- 9210130 TI - Dysplasia epiphysealis hemimelica. AB - Dysplasia epiphysealis hemimelica is a rare developmental disorder affecting one or more epiphyses in a limb or a tarsal or carpal bone. It is due to abnormal cartilage proliferation with subsequent endochondral ossification and generally involves the medial or lateral half of the epiphysis. Our case involved the lateral aspect of the distal femoral epiphysis, which was managed with local resection. PMID- 9210131 TI - Acute compartment syndrome in ruptured Baker's cyst. AB - We present a case report of a previously healthy 43-year-old man with a Baker's cyst and spontaneous venous bleeding in a leg compartment, which caused a compartment syndrome. A thorough evaluation of this case and the treatment are explored. PMID- 9210132 TI - Intracapsular chondroma. PMID- 9210133 TI - Radioligand-binding methods for membrane preparations and intact cells. PMID- 9210134 TI - Site-directed mutagenesis and chimeric receptors in the study of receptor-ligand binding. PMID- 9210135 TI - Approaches to the stable transfection of G protein-coupled receptors. PMID- 9210136 TI - Methods for transient expression of hetero-oligomeric ligand-gated ion channels. PMID- 9210137 TI - The generation of receptor-selective antibodies. PMID- 9210138 TI - In situ hybridization. PMID- 9210139 TI - Immunocytochemical methods for investigating receptor localization. PMID- 9210140 TI - Measurement of agonist-stimulated [35S]GTP gamma S binding to cell membranes. PMID- 9210141 TI - Autoradiographic visualization in brain of receptor-G protein coupling using [35S]GTP gamma S binding. PMID- 9210142 TI - Agonist-induced high-affinity GTP hydrolysis as an index of receptor-mediated G protein activation in mammalian brain membranes. PMID- 9210143 TI - Use of random-saturation mutagenesis to study receptor-G protein coupling. PMID- 9210144 TI - Identification and quantitation of G protein alpha-subunits. PMID- 9210145 TI - Covalent modification of G proteins by affinity labeling. PMID- 9210146 TI - Heterologous expression of receptors and signaling proteins in adult mammalian sympathetic neurons by microinjection. PMID- 9210148 TI - Use of antisense-generating plasmids to probe the function of signal transduction proteins in primary neurons. PMID- 9210147 TI - Nuclear application of antisense oligonucleotides by microinjection and ballistomagnetic transfer to identify G protein heterotrimers activating phospholipase C. PMID- 9210149 TI - Protocols employed in the investigation of G protein-coupled receptor phosphorylation. PMID- 9210151 TI - Radioligand binding measurement of receptor sequestration in intact and permeabilized cells. PMID- 9210150 TI - Assay of G protein-coupled receptor kinase activity by rhodopsin phosphorylation. PMID- 9210152 TI - Regulation of receptor expression. Analysis of receptor mRNA and gene transcription. PMID- 9210153 TI - DNA typing in forensic medicine and in criminal investigations: a current survey. AB - Since 1985 DNA typing of biological material has become one of the most powerful tools for personal identification in forensic medicine and in criminal investigations [1-6]. Classical DNA "fingerprinting" is increasingly being replaced by polymerase chain reaction (PCR) based technology which detects very short polymorphic stretches of DNA [7-15]. DNA loci which forensic scientists study do not code for proteins, and they are spread over the whole genome [16, 17]. These loci are neutral, and few provide any information about individuals except for their identity. Minute amounts of biological material are sufficient for DNA typing. Many European countries are beginning to establish databases to store DNA profiles of crime scenes and known offenders. A brief overview is given of past and present DNA typing and the establishment of forensic DNA databases in Europe. PMID- 9210154 TI - Functional differences of myosin heavy-chain isoforms in skeletal muscle. PMID- 9210155 TI - Vertical migration of Chernobyl-derived radiocesium in Bavarian grassland soils. PMID- 9210156 TI - The economics of health promotion. PMID- 9210157 TI - The preferable future for nursing. PMID- 9210158 TI - American Board of Nursing Specialties: past, present, and future. PMID- 9210159 TI - Equitable access to cancer services in the 21st century. PMID- 9210160 TI - "To be of use": enhancing the utility of qualitative research. PMID- 9210161 TI - The Teaching Nursing Home Program: enduring educational outcomes. PMID- 9210162 TI - Interaction of alpha s2- and beta-casein signal peptides with DMPC and DMPG liposomes. AB - Interactions of casein signal peptides (CSP) and derivatives were detected with dimyristoylphosphatidyl-glycerol and -choline liposomes. Fluorescence anisotrophy indicated that the peptides interact better with DMPG than DMPC, inserting at a limited depth in the bilayer. Stronger interaction was detected for derivatives of beta-CSP than of alpha s2-CSP. Tryptophan fluorescence (intrinsic, energy transfer, quenching) showed that the central hydrophobic core of CSP was buried in the bilayer whereas both ends remained outside, adopting a hairpin-like conformation. The secondary structure of the CSP was not affected by their interactions with phospholipids. beta-CSP derivatives show both lytic and fusogenic activities. PMID- 9210163 TI - Isolation and structural elucidation of two pyrokinins from the retrocerebral complex of the American cockroach. AB - By monitoring the contractile activity of the hyperneural muscle of the American cockroach in vitro two peptides were isolated from the retrocerebral complex of the American cockroach. Three purification steps using reversed-phase high performance liquid chromatography on C-18 columns containing trifluoroacetic acid or heptafluorobutyric acid as organic modifiers were sufficient to achieve homogeneous peptide preparations. The structures of both peptides were elucidated by a combination of Edman degradation and mass spectrometry which yielded the following structures: His-Thr-Ala-Gly Phe-Ile-Pro-Arg-Leu-NH2 (Pea-PK-1) and Ser Pro-Pro-Phe-Ala-Pro-Arg-Leu-NH2 (Pea-PK-2). The C-terminal sequence Phe-X-Pro-Arg Leu-NH2 characterized the peptides as members of the insect pyrokinin family. The synthetic peptides were shown to have the same retention times as the natural peptides. The occurrence of both peptides in the retrocerebral complex suggests a physiological role as neurohormones. The effects of the synthetic pyrokinis were clearly distinguishable in their actions on the hyperneural muscle. Regarding the threshold concentrations, Pea-PK-2 was only 0.3% as active as Pea-PK-1. PMID- 9210164 TI - Amino acid sequences and activities of multiple hyperglycemic hormones from the Kuruma prawn, Penaeus japonicus. AB - By means of a single-step reversed-phase HPLC, six CHH family peptides were isolated from sinus gland extracts of the kuruma prawn, Penaeus japonicus. Five of these peptides (Pej-SGP-I, -II, -III, -V, and -VI) expressed hyperglycemic activity and were considered to be the hyperglycemic hormones of this prawn. The amino acid sequences of the five peptides were determined (the amino acid sequence of Pej-SGP-III had been previously determined), revealing that all consisted of 72 amino acid residues with a free amino-terminus and an amidated carboxyl-terminus. They showed considerable sequence similarity to one another, but much less similarity to Pej-SGP-IV with molt-inhibiting activity, the sequence of which had also been determined previously. Examination of the dose response relationship of these peptides showed that they were equally potent but had different efficacies, which were in the order of Pej-SGP-V, -VI > -III, -I > II, corresponding well with their sequence characteristics. PMID- 9210165 TI - Neuropeptide K and neurokinin A stimulate CRH and ACTH release by rat adrenal medulla in vitro. AB - Tachykinins are a family of peptides that are able to modulate the activity of the hypothalamo-pituitary CRH-ACTH system. Mammalian tachykinins include neurokinin A (NKA), neurokinin B (NKB), neuropeptide K (NPK), and substance P (SP). We investigated by RIA the effects of tachykinins on the release of CRH and ACTH by rat adrenal medulla in vitro. NKA and NPK concentration-dependently enhanced the release of both CRH and ACTH, NPK being more active than NKA. NKB exerted only a minor stimulatory action exclusively on CRH release, and SP was ineffective. The stimulatory effect of both NKA and NPK on ACTH release was blocked by the CRH receptor antagonist alpha-helical-CRH, thereby suggesting that the increase in ACTH secretion is consequent to the stimulation of CRH release. These findings indicate that NKA and NPK are stimulators not only of the central (hypothalamo-pituitary), but also of the peripheral (intramedullary) branch of the CRH-ACTH system. PMID- 9210166 TI - Effects of captopril on responses to bradykinin in the hindquarters vascular bed of the rat. AB - The effects of captopril, and angiotensin converting enzyme (ACE) inhibitor, on responses to bradykinin (BK), to angiotensin (Ang) I and II, and to other agonists were investigated in the hindquarters vascular bed of the rat. Under conditions of controlled flow, intra-arterial (i.a.) injections of BK in doses of 0.1-1.0 microgram, produced dose related decreases in hindquarters perfusion pressure and evoked decreases in systemic arterial pressure. Intra-arterial injections of Ang I and II produced dose-related increases in hindquarters perfusion pressure. Following administration of captopril in a dose of 2 mg/kg i.v., vasodilator responses to i.a. injections of BK were only slightly enhanced in the hindquarters vascular bed, whereas the evoked systemic vasodepressor responses to i.a. injections of BK were markedly enhanced by the ACE inhibitor. Captopril significantly reduced vasoconstrictor responses to i.a. injections of Ang I, whereas vasoconstrictor responses to i.a. injections of Ang II were significantly enhanced. The ACE inhibitor did not significantly alter vasodilator responses to i.a. injections of acetylcholine, nitroglycerin, nitric oxide, albuterol, or pinacidil. The present data show that BK has potent vasodilator activity in the hindquarters vascular bed of the rat and suggest that the site of action is most likely upstream from the site of inactivation, whereas the site of action of Ang I is at or near the site of conversion to Ang II in the hindquarters vascular bed. The observation that the evoked systemic vasodepressor responses to i.a. injections of BK were greatly enhanced, suggest that the lung is the major site of inactivation of BK. PMID- 9210168 TI - Solid-phase synthesis of hydroxyethylamine angiotensin analogues. AB - Three hydroxyethylamine analogues of angiotensins II, III, and IV were prepared by solid-phase methods. The resin-bound peptide was alkylated with the iodomethylketone derivative of the N-terminal amino acid, followed by reduction to the alcohol using sodium borohydride. The iodomethylketones can be made in good yields from commercially available N-protected amino acids. The compounds were evaluated for their ability to displace labeled angiotensins from bovine adrenal membranes, and their metabolic stability tested in kidney homogenates and aminopeptidase M preparations. The hydroxyethylamine amide bond replacement reduced the affinity of the analogues; however, they were substantially more stable to enzymatic degradation. PMID- 9210167 TI - Angiotensinogen and natriuretic peptide mRNAs in rat brain: localization and differential regulation by adrenal steroids in hypothalamus. AB - Adrenal steroids have been shown to modulate angiotensin II and natriuretic peptide systems--peptide synthesis and metabolism--in vitro. In the present study the effects of adrenal steroids on mRNA encoding the angiotensin II precursor, angiotensinogen (AOGEN), and the natriuretic peptides, atrial natriuretic peptide (ANP) and C-type natriuretic peptide (CNP) in the rat hypothalamus were investigated using quantitative in situ hybridization histochemistry of [35S]- and [33P]-labeled oligonucleotide probes. Adrenalectomy produced an apparent overall decrease in preproAOGEN (ppAOGEN) mRNA in presumed astrocytes in the anterior hypothalamus with significant decreases (ANOVA) measured in the medial preoptic area, the ventral region of the medical preoptic area, the paraventricular, suprachiasmatic, supraoptic, and periventricular nuclei. ppAOGEN mRNA levels were restored by both glucocorticoid (dexamethasone; 2 micrograms/ml in drinking water) and mineralocorticoid (aldosterone; 50 micrograms/kg, SC) replacement. Treatment of intact animals with dexamethasone (2 micrograms/ml in drinking water for 5 days) and aldosterone (100 micrograms/kg, SC, daily for 10 days) produced a significant increase in ppAOGEN mRNA in those hypothalamic regions affected by adrenalectomy. ppANP and ppCNP mRNA-positive neurons were successfully detected using [35S]- and [33P]-labeled probes, respectively, and were abundant in the anterior hypothalamus, particularly in the anteromedial preoptic nucleus of the medial preoptic area. In contrast to the effects on ppAOGEN mRNA, however, alterations in adrenal steroid levels did not significantly change ppANP or ppCNP mRNA levels in neurons of the anteromedial preoptic nucleus or in the arcuate nucleus. These results indicate that adrenal steroids modulate AOGEN gene transcription in vivo, consistent with previous reports of increased levels of ppAOGEN mRNA in a number of brain regions in response to acute dexamethasone treatment and reports of decreased AOGEN immunoreactivity in brain regions of adrenalectomized rats. In contrast, despite reports of modulation of hypothalamic ANP immunoreactivity following adrenalectomy and dexamethasone treatment, it would appear that adrenal steroids do not alter the transcription or stability of hypothalamic natriuretic peptides mRNA in vivo. PMID- 9210169 TI - Tone-dependent vasodilator responses to proadrenomedullin NH2-terminal 20 peptide in the hindquarters vascular bed of the rat. AB - Responses to proadrenomedullin NH2-terminal 20 peptide (PAMP) were investigated in the systemic and hindquarters vascular bed of the rat. Intravenous injections of PAMP and adrenomedullin (ADM) produced dose-related decreases in systemic arterial and hindquarters perfusion pressure, which were not altered by alpha receptor or adrenergic nerve terminal blocking agents. PAMP was 100-fold less potent than ADM, and hindquarters vasodilator responses to both peptides were similar in innervated and denervated preparations. When baseline tone was increased with phenylephrine and U46619 or decreased with sodium nitroprusside, vasodilator responses to PAMP and ADM were correlated with the basal level of tone, suggesting that responses to both peptides are dependent on the baseline level of vasoconstrictor tone in the hindquarters vascular bed of the rat. PMID- 9210170 TI - High affinity binding of 125I-neurotensin to dispersed cells from chicken liver and brain. AB - Dispersed cells from chicken brain and liver were found to possess cell surface binding sites for 125I-neurotensin (125I-NT). Scatchard analyses indicated the presence of high affinity (K4, 25-80 pM) and low affinity (Kd, 250-450 pM) components in adult tissues. Binding capacity was reduced 25-40% by incubation with pertussis toxin. Ontogenetic studies indicated that NT receptor capacity increased approximately 20-fold from the embryonic stage to adult. Cross-linking of 125I-NT to intact cells labeled one major band (52 kDa, > or = 90%) and two minor bands (40 and 90 kDa, < or = 10%) which could represent distinct NT receptors or one receptor partly degraded or cross-linked to G-protein(s). The binding of 125I-NT to dispersed cells was enhanced by reduction with dithoithreitol and suppressed by alkylation with N-ethyl-maleimide (NEM), maleimidocaproic acid (MCA) and p-chloromercuribenzenesulfonate (PCMBS). Since MCA and PCMBS do not permeate cells, this suggests that the sulfhydryl group(s) critical to binding are located within the NT receptor itself. Preincubation of cells with NT prior to treatment with NEM diminished its inhibitory effect, suggesting that the critical SH-group(s) were within the NT binding pocket or were protected by an allosteric effect. These results suggest that one or more of the nine cysteine residues in the NT receptor is involved in the NT binding reaction. PMID- 9210171 TI - EEDQ reduces the striatal neurotensin mRNA response to haloperidol. AB - Haloperidol is believed to induce neurotensin/neuromedin N (NT/ N) gene expression in the dorsolateral striatum (DLST) of the rat brain via dopamine D2 receptor blockade, but is also known to interact with other receptors as well. To further characterize haloperidol's effects, rats were treated with the irreversible monoaminergic receptor antagonist N-ethoxycarbonyl-2-ethoxy-1,2 hydroxyquinolone (EEDQ). In situ hybridization was performed for NT/N mRNA. D2 like and sigma receptor autoradiography was performed using 125I-sulpride and 3H 1,3-di-o-tolylguanidine (DTG), respectively. Despite antagonism of D2 receptors, pretreatment with EEDQ failed to significantly reduce the NT/N mRNA response when given 3 days prior to the haloperidol challenge. These data suggest that the acute effects of haloperidol on NT/N mRNA expression in large part result from D2 receptor antagonism. Nonetheless, a contribution of other receptors can not be excluded. PMID- 9210172 TI - Effects of amylin on human osteoblast-like cells. AB - Amylin has been reported to have bone-conserving effects. In the present study we evaluated the possible activity of the peptide on human osteoblast-like (hOB) cells in primary culture. Amylin between 10(-9) and 10(-6) M, dose-dependently stimulated cell proliferation with a maximal effect (200%) at 10(-6) M. In addition, amylin increased osteocalcin production when hOB cells were exposed to 1,25(OH)2D3 (10(-8)M) but there was a nonsignificant upward trend on alkaline phosphatase activity. The present results suggest that amylin could be included among the group of peptides endowed with osteogenic activity. PMID- 9210173 TI - Effects of cholecystokinin agonists on striatal neurons are reduced by acetylcholine. AB - The present study investigated whether acetylcholine, a transmitter of striatal interneurons, modulates responses of neostriatal neurons to agonists of the neuropeptide cholecystokinin (CCK). Single unit activity was recorded in rats anesthetized with urethane. Acetylcholine and CCK agonists (the CCKA receptor agonists A-71378 and A-71623; the CCKB receptor agonist Suc-CCK-4) were iontophoretically administered alone and in combination. The CCK agonists excited about one third of the neurons. The excitatory effects of both the CCKB and the CCKA receptor agonists were mainly reduced or changed to suppression of activity by acetylcholine (Wilcoxon test p < 0.001). Atropine did not significantly change the neuronal responses to the CCK agonists. The suppressive action of acetylcholine could be diminished by additional administration of atropine. The results suggest that the modulatory action of cholecystokinin does not only depend on the actual state of excitability in striatal neurons, but could be changed by acetylcholine released from interneurons. PMID- 9210174 TI - Endogenous CCK in the control of gastric emptying of glucose and maltose. AB - A role for endogenous cholecystokinin (CCK) in the control of gastric emptying of liquid glucose and maltose test meals in rhesus monkeys was assessed. Intragastric administration of a dose range (10-100 micrograms/kg) of the CCKA receptor antagonist devazepide produced a dose dependent acceleration of the emptying of 100 ml 300 mOsm test meals of glucose and maltose but had no effect on the emptying of a hyperosmotic (750 mOsm) NaCl solution. At the 100 micrograms/kg dose, the emptying of glucose and maltose meals were as rapid as the emptying of physiological NaCl. These data expand the demonstrated role of endogenous CCK in the slowing of gastric emptying of nutrients in rhesus monkeys to carbohydrates and suggest that previous negative results were due to the hyperosmotic nature of the glucose solutions. PMID- 9210175 TI - Several receptors mediate the antisecretory effect of peptide YY, neuropeptide Y, and pancreatic polypeptide on VIP-induced fluid secretion in the rat jejunum in vivo. AB - Several Y receptor subtypes have been cloned and/or pharmacologically characterized that mediate the effects of the regulatory peptides peptide YY (PYY), neuropeptide Y (NPY), and pancreatic polypeptide (PP). These peptides possess antisecretory properties on the intestine. This effect can be blocked in vivo by neural antagonists, suggesting the intervention of neural receptors, although epithelial PYY-preferring receptors have been evidenced on jejunal crypt cells. The purpose of the present experiments was to compare the antisecretory properties in vivo of a series of PYY and NPY derivatives with various affinities for different Y receptor subtypes, in order to determine which subtypes were involved. A model of VIP-stimulated secretion by rat jejunal loops was used. The results were compared with the binding affinities for PYY-preferring receptors determined on rat jejunal crypt cell membranes. Full-length PYY(1-36) was about three times more potent than NPY(1-36), and 10 times more potent than PP in the low dose range. PP, however, had a low efficacy limited to about 50% inhibition of VIP effect. Both Y1 agonists ([Leu31, Pro34]PYY and [Leu31,Pro34]NPY), and Y2 agonists [C-terminal fragments ranging from PYY (3-36) and NPY(3-36) to PYY(22 36) to NPY(22-36)] displayed potent antisecretory properties. PYY derivatives and fragments were always more potent than their respective NPY counterparts. In contrast, Y1 derivatives and PP had very low affinity for the epithelial PYY receptor as measured in vitro by radioreceptor assay. These data suggest that the antisecretory effect of PYY/NPY/PP peptides in vivo involves the effects of several receptors: a Y2-like, PYY-preferring receptor identical to the epithelial receptor, a Y1-like receptor, and a third receptor with high affinity for PP. PMID- 9210176 TI - Differential responsiveness of proximal and distal colonic mucosa to gastrin. AB - In vivo and in vitro experiments were performed to examine the responsiveness of the proximal and distal colonic mucosa to the growth-promoting action of gastrin. Infusion (osmotic minipump) of gastrin G-17-I (250 ng/kg/h) for 5 days to 4-month old male Fischer-344 rats resulted in a significant (90-150%) increase in proliferative activity (as assessed by BrdU or PCNA immunoreactivity) in the distal colonic mucosa. In contrast, gastrin caused no apparent change in proliferative activity in the proximal colon. Because tyrosine kinases (Tyr-ks) are thought to be critically involved in regulating the trophic action of gastrin, responsiveness of isolated colonocytes from both segments of the colon to gastrin (1 x 10(-9) M) was also examined. Exposure of isolated colonocytes from the distal, but not from the proximal, colon to gastrin for 2 min resulted in a significant (73%) stimulation in Tyr-k activity. This was also accompanied by a marked rise in phosphorylation of at least six membrane proteins with M, of 55, 60, 70, 94, and 170 kDa. Tyr-k activity induced by gastrin in colonocytes from the distal colon was inhibited by tyrphostin (3.2 microM) but not by staurosporine (20 nM). In colonocytes from the distal colon, gastrin also stimulated phospholipase C (PLC) activity, which could also be inhibited by tyrphostin, but not by staurosporine. We conclude that mucosa of the distal, but not the proximal, colon responds to the trophic action of gastrin. Tyr-ks are thought to be involved in the regulation of this process. PMID- 9210177 TI - Endoproteolysis at tetrabasic amino acid sites in procalcitonin gene-related peptide by pituitary cell lines. AB - The specificity of neuroendocrine prohormone convertases for tetrabasic amino acid sites was investigated. Mutations were introduced into the tetrabasic cleavage site of the procalcitonin gene-related peptide (proCGRP) cDNA and these mutated cDNA's were expressed in AtT-20 cells which predominantly express the endoprotease prohormone convertase-1 (PC1/3), and in GH3 cells which predominantly express prohormone convertase-2 (PC2). Mutations were introduced into the proCGRP cDNA which converted the wild-type ArgArgArgArg site to LysLysArgArg and ArgArgLysLys, and the proCGRP variants were stably transfected into AtT-20 and GH3 cells. ProCGRP containing each of the LysLysArgArg permutations were efficiently cleaved in both AtT-20 and GH3 cells. Cleavage of LysLysArgArg in exogenous proCGRP, but not in endogenous POMC, suggests that the specificity of cleavage at tetrabasic sites is not defined solely by the endoproteases expressed by the cell or by the amino acid sequence at the cleavage site, but is also dependent on the structure of the propeptide. PMID- 9210178 TI - Neuropeptide FF receptors of mouse olfactory bulb: binding properties and stimulation of adenylate cyclase activity. AB - Neuropeptide FF (NPFF) receptors have been characterized in mouse olfactory bulb membranes by using [125I][1DMe]Y8Fa. The specific binding of this NPFF analogue was time and concentration dependent, reversible, saturable, and of high affinity (Kd = 0.022 nM, Bmax = 56.4 fmol/mg protein). In olfactory bulb membranes, NaCl increased the affinity of [125I][1DMe]Y8Fa by decreasing the dissociation rate constant (k-1). In contrast, the nonhydrolyzable analogue of GTP, Gpp[NH]p, decreased the maximal number of binding sites suggesting a coupling of NPFF receptors to a G-protein. In mouse olfactory bulb and spinal cord membranes, NPFF analogues stimulated adenylate cyclase activity in a time- and dose-dependent manner, whereas in the cerebellum, which does not possess NPFF receptors, low cAMP production was stimulated by NPFF. Our data are consistent with guanine nucleotide binding protein regulation of NPFF receptors positively coupled to adenylate cyclase. PMID- 9210179 TI - Effects of the neuropeptide thyrotropin-releasing hormone on GABAergic synaptic transmission of CA1 neurons of the rat hippocampal slice during hypoxia. AB - Because thyrotropin-releasing hormone (TRH) has been suggested to improve recovery of brain neurons from hypoxia, which strongly impairs GABAergic synaptic transmission, the present electrophysiological study used intracellular recording from CA1 neurons of the rat hippocampal slice to examine the cellular mechanisms underlying this phenomenon. Hypoxia induced by superfusion with a medium devoid of oxygen evoked typical membrane hyperpolarization, fall in input resistance, and strong depression of monosynaptic, GABAA receptor-mediated fast inhibitory postsynaptic potentials (IPSPs). The depression of fast IPSPs during hypoxia was found to be due to a combination of factors such as shift in the IPSP reversal potential and membrane hyperpolarization. GABAB receptor-mediated slow IPSPs were comparatively less sensitive to hypoxia. TRH (10 microM), applied 1 min prior to hypoxia, selectively accelerated recovery of membrane potential and delayed return of fast IPSPs to control amplitude without changing the mechanisms responsible for depression of GABAergic transmission. In conclusion, despite a slower recovery of IPSPs, TRH facilitated earlier return of neuronal excitability after the hypoxic period. PMID- 9210180 TI - Motilin and clinical application. AB - Motilin is a regulatory polypeptide of 22 amino acid residues and orginates in motilin cells scattered in the duodenal epithelium of most mammals and chickens. Motilin is released into the general circulation at about 100-min intervals during the interdigestive state and is the most important factor in controlling the interdigestive migrating contractions. Recent studies have revealed that motilin stimulates endogenous release of the endocrine pancreas. Clinical application of motilin as a prokinetic has become possible since erythromycin and its derivatives were proved to be nonpeptide motilin agonists. PMID- 9210181 TI - A structure-activity analysis of the cloned rat and human Y4 receptors for pancreatic polypeptide. AB - We cloned and expressed the rat Y4 receptor for pancreatic polypeptide (PP). Structure-activity profiles derived from 125I-PP binding assays and [cAMP] radioimmunoassays reveal a selective receptor interaction with rat PP vs. neuropeptide Y (NPY) or peptide YY (PYY). Rat and human Y4 receptor clones share 75% amino acid identity. Based on [cAMP] radioimmunoassay, the human Y4 receptor exhibits a less selective interaction with rat PP vs. NPY or PYY and a greater dependence on N-terminal PP residues, relative to rat Y4. Differences in sequence and structure-activity profiles suggest the rat be used with caution to model human Y4 receptor function. PMID- 9210182 TI - A study of calcitonin effect on synaptosomal membrane enzymes. AB - Calcitonin is a hormone peptide produced by the thyroid gland, whose best described role is to prevent bone reabsorption. It also participates in other biological functions, even at central nervous system level. We studied the effect of added calcitonin on ATPase and acetylcholinesterase activities in synaptosomal membranes isolated from rat cerebral cortex. Calcitonin at 10(-7) - 10(-5)M concentration decreased 20-40% Na+, K(+)-ATPase and 15-25% K(+)-p nitrophenylphosphatase activities, and at 10(-6)-10(-5)M reduced 20-30% Mg(2+)-p nitrophenylphosphatase activity. However, this peptide failed to modify Mg(2+) - and Ca(2+)-ATPase or acetylcholinesterase activities. Results suggest that the sodium pump may be a target for calcitonin effects at neuronal level. Thus, calcitonin inhibition of sodium/potassium transport through synaptic membranes supports a regulatory role of this peptide on neurotransmission. PMID- 9210184 TI - Aerosol electrostatics. I: Properties of fine powders before and after aerosolization by dry powder inhalers. AB - PURPOSE: To evaluate the dependence of fine particle dose charge (FPD charge) generated from powder inhalers on physico-chemical properties of the inhalation powder, inhaler type, deaggregation mechanism, dose number and/or retained powder. METHODS: Electrostatic charges were determined on micronized powders and aerosolized fine particle doses withdrawn from two, high efficiency, multidose powder inhalers, Turbohaler and prototype Dryhaler. The behavior of terbutaline sulfate, budesonide, albuterol (sulfate and base), beclomethasone dipropionate and lactose was assessed before and after aerosolization. RESULTS: Both inhalers conferred triboelectric FPD charges during aerosolization in the range -400 pC through +200 pC. Specific charges (charge/unit mass) on the fine particle doses of budesonide from Dryhaler were significantly less than those from Turbohaler (p < 0.01). Electrostatic charges on the potentially respirable cloud of terbutaline sulfate generated by Bricanyl Turbohaler were positive and/or negative and unpredictable. With Pulmicort Turbohaler, FPD charges on budesonide were always positive. Dryhaler was used to determine the chemical dependence of fine particle triboelectrification during the aerosolization of pure materials. A triboelectric series was constructed from the Dryhaler results ranking the powders from positive to negative as budesonide > lactose > albuterol sulfate > terbutaline sulfate > or = albuterol > or = beclomethasone dipropionate. CONCLUSIONS: While there was no evidence of FPD charge dependence upon dose number with either inhaler, FPD charges were dependent upon the powder under investigation, as well as the construction and deaggregation mechanism of the inhaler. The specific charge on the fine particle dose of budesonide from Turbohaler corresponded to approximately 200 electronic charges per particle, a value which is known to affect both total and regional aerosol deposition in the human lung. Electrostatic charge effects may be important determinants of aerosol behavior and should not be neglected. PMID- 9210185 TI - Cellular delivery of nucleoside diphosphates: a prodrug approach. AB - PURPOSE: This study is concerned with cellular delivery/generation of 2'-azido-2' deoxyuridine and -deoxycytidine diphosphate (N3UDP or N3CDP), potent inhibitors of ribonucleotide reductase. It characterizes the phosphorylation steps involved in the conversion of 2'-azido-2'-deoxyuridine (N3Urd) and 2'-azido-2' deoxycytidine (N3Cyd) to the corresponding diphosphates and explores a prodrug approach in cellular delivery of the inhibitor which circumvents the requirement of deoxynucleoside kinases. METHODS: Cell growth of CHO and 3T6 cells of known deoxycytidine kinase level was determined in the presence of N3Urd and N3Cyd. Activity of ribonucleotide reductase was determined in the presence of the azidonucleosides as well as their mono- or di-phosphates in a Tween 80-containing permeabilizing buffer. A prodrug of 5'-monophosphate of N3Urd was prepared and its biological activity was evaluated with CHO cells as well as with cells transfected with deoxycytidine kinase. RESULTS: N3Urd failed to inhibit the growth of both cell lines, while N3Cyd was active against 3T6 cells and moderately active against CHO cells. These results correlate with the deoxycytidine kinase levels found in the cells. Importance of the kinase was further established with the finding that the nucleoside analogs were inactive as reductase inhibitors in a permeabilized cell assay system while their mono- and di-phosphates were equally active. The prodrug was active in cell growth inhibition regardless of the deoxycytidine kinase level. CONCLUSIONS: The azidonucleosides become potent inhibitors of the reductase by two sequential phosphorylation steps. The present study indicates that the first step to monophosphate is rate-limiting, justifying a prodrug approach with the monophosphate. PMID- 9210186 TI - Dynorphin peptides: antagonists of melanocortin receptors. AB - PURPOSE: To identify possible targets that mediate the non-opioid effects of dynorphin-A (DynA), effects that include inflammation and aggravation of traumatic nerve injury. METHOD: We examined dynorphin peptides for functional interaction with the closely related melanocortin (MC) system. RESULTS: DynA-(1 13)NH2 and other related opioid dynorphin peptides antagonize the human MC1, MC3 and MC4 receptors, and an amphibian MC receptor, with dissociation constants (Kd's) of 40 to 150 nM. The affinity of dynorphin's interaction with MC receptors is therefore greater than with other previously proposed non-opioid targets of dynorphin, which require micromolar concentrations. Dynorphin also antagonizes the adrenocorticotropic hormone (ACTH; MC2) receptor and an MC-like receptor endogenous to COS-7 cells, but with lower efficacy. In contrast DynA had no effect on seven control receptors and was only weakly effective at two others. Metabolites of dynorphin derived from cleavage of the amino terminal Tyr residue, such as DynA(2-17), lack opioid activity yet still produce a number of well established non-opioid effects. These des-Tyr derivatives also antagonized each of the five MC receptors examined. CONCLUSIONS: DynA peptides were found to antagonize MC receptors in vitro with potencies that parallel those reported for pharmacological non-opioid effects of dynorphins in vivo. The combination of DynA and its active metabolites may reach levels sufficient to inhibit MC receptors physiologically. Dynorphin inhibition of MC receptors could prove to be an example of crosstalk between two distinct yet phylogenetically related neurotransmitter systems. PMID- 9210183 TI - Electrically-assisted transdermal drug delivery. AB - Electrically-assisted transdermal delivery (EATDD) is the facilitated transport of compounds across the skin using an electromotive force. It has been extensively explored as a potential means for delivering peptides and other hydrophilic, acid-labile or orally unstable products of biotechnology. The predominant mechanism for delivery is iontophoresis, although electroosmosis and electroporation have also been investigated. The focus of this review is to put these different mechanisms in perspective and relate them to the drug and skin model system being investigated. PMID- 9210187 TI - Basic studies for the practical use of bitterness inhibitors: selective inhibition of bitterness by phospholipids. AB - PURPOSE: We examined the effects of phospholipids such as phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol (PI), and phosphatidic acid (PA) on human taste sensation to various substances. METHODS: The effects were evaluated psychophysically using paid volunteers. RESULTS: PA inhibited the bitterness of various substances dissolved in water without affecting sweetness, saltiness, and sourness, although its inhibitory activity was less than that of PA-LG. PI also showed inhibitory activity on bitterness, although its activity was less than PA. A soybean lecithin fraction containing high contents of PA and PI also demonstrated inhibitory activity on the bitterness of various substances. Both the incorporation of either PA or the lecithin fraction into granules containing quinine and the coating of the granules with PA or the fraction effectively inhibited the bitterness of quinine. CONCLUSIONS: The lecithin fraction is permitted for use as an additive to drugs and food and can be produced on an industrial scale. It is expected that the lecithin fraction will be used safely as a bitterness inhibitor for practical applications. PMID- 9210188 TI - The effect of benzyl alcohol on recombinant human interferon-gamma. AB - PURPOSE: The goal of this study was to investigate the conformational change and aggregation of recombinant human interferon-gamma (rhIFN-gamma) as a result of interaction between benzyl alcohol and the protein. The effects of buffer concentration, buffer species, ionic strength, rhIFN-gamma and benzyl alcohol concentrations on the dynamics of the interaction in liquid formulations were also examined. METHODS: The effect of benzyl alcohol on the secondary and tertiary structure of rhIFN-gamma in succinate and acetate buffers was studied using far-UV and near-UV circular dichroism spectrophotometry, respectively. Dynamic light scattering was employed to detect aggregate formation due to the interaction of benzyl alcohol with rhIFN-gamma. RESULTS: The addition of benzyl alcohol at 0.9% (w/v) in various liquid rhIFN-gamma formulations induced changes in circular dichroism (CD) spectra of the protein in the near-UV region, while the CD spectra in the far-UV region remained unaltered. There were gradual decreases in ellipticity with time throughout the near-UV CD spectra. The decreases in near-UV ellipticity induced by benzyl alcohol were accompanied by the formation of high molecular weight aggregates as measured by dynamic light scattering. Loss in near-UV ellipticity was accelerated at lower protein concentration and by increasing buffer or benzyl alcohol concentration. It was also faster in succinate than in acetate buffer. Formulation ionic strength did not affect the CD spectral changes in both the near- and far-UV regions. CONCLUSIONS: Interaction between benzyl alcohol and rhIFN-gamma is formulation dependent. Protein concentration, buffer species, buffer concentration, and preservative concentration play a significant role in determining the extent of the interaction and consequently the stability of the product. PMID- 9210190 TI - Dependence of the molecular mobility and protein stability of freeze-dried gamma globulin formulations on the molecular weight of dextran. AB - PURPOSE: The effect of the molecular weight of dextran on the molecular mobility and protein stability of freeze-dried serum gamma-globulin (BGG) formulations was studied. The stabilizing effect of higher molecular weight dextran is discussed in relation to the molecular mobility of the formulations. METHODS: The molecular mobility of freeze-dried BGG formulations containing dextrans of various molecular weights was determined based on the free induction decay of dextran and water protons measured by proton NMR. The protein stability of the formulations was determined at temperatures ranging from 20 to 70 degrees C by size exclusion chromatography. RESULTS: Changes in the molecular mobility of freeze-dried formulations that occurred at temperatures below the glass transition temperature could be detected as the molecular mobility-changing temperature (Tmc), at which dextran protons started to exhibit a Lorentzian relaxation decay due to higher mobility in addition to a Gaussian relaxation decay. Tmc increased as the molecular weight of dextran increased. The proportion of dextran protons which exhibited the higher mobility relaxation process (Phm) at temperatures above Tmc decreased as the molecular weight of dextran increased. Protein stability was closely related to molecular mobility. The temperature dependence of the denaturation rate changed at around Tmc, and denaturation in the microscopically liquidized state decreased as Phm decreased with increasing molecular weight of dextran. CONCLUSIONS: The effect of the molecular weight of dextran on the protein stability of freeze-dried BGG formulations could be explained in terms of the parameters obtained by 1H-NMR such as Tmc and Phm. These parameters appear to be useful in preformulation and stability prediction of freeze-dried formulations. PMID- 9210189 TI - The stabilization and encapsulation of human growth hormone into biodegradable microspheres. AB - PURPOSE: To produce and evaluate sustained-acting formulations of recombinant human growth hormone (rhGH) made by a novel microencapsulation process. METHODS: The protein was stabilized by forming an insoluble complex with zinc and encapsulated into microspheres of poly (D,L-lactide co-glycolide) (PLGA) which differed in polymer molecular weight (8-31 kD), polymer end group, and zinc content. The encapsulation procedure was cryogenic, non-aqueous, and did not utilize surfactants or emulsification. The rhGH extracted from each of these microsphere formulations was analyzed by size-exclusion, ion-exchange and reversed-phase chromatography, SDS-polyacrylamide gel electrophoresis, peptide mapping, and cell proliferation of a cell line expressing the hGH receptor. In addition, the in vivo release profile was determined after subcutaneous administration of the microspheres to rats and juvenile rhesus monkeys. RESULTS: Protein and bioactivity analyses of the rhGH extracted from three different microsphere formulations showed that the encapsulated protein was unaltered relative to the protein before encapsulation. In vivo, microsphere administration to rats or monkeys induced elevated levels of serum rhGH for up to one month, more than 20-fold longer than was induced by the same amount of protein injected subcutaneously as a solution. The rate of protein release differed between the three microsphere formulations and was determined by the molecular weight and hydrophobicity of the PLGA. The serum rhGH profile, after three sequential monthly doses of the one formulation examined, was reproducible and showed no dose accumulation. CONCLUSIONS: Using a novel process, rhGH can be stabilized and encapsulated in a solid state into PLGA microspheres and released with unaltered properties at different rates. PMID- 9210191 TI - In vivo gene transfer by intravenous administration of stable cationic lipid/DNA complex. AB - PURPOSE: A stable cationic lipid/DNA complex has been developed for in vivo gene transfer. The formulation capitalizes on a previously described procedure to obtain stable lipid/DNA complexes for in vitro gene transfer (1). METHODS: Conditions for DNA/lipid complex formation were modified to yield a DNA concentration of 1 mg/ml. Heat stable alkaline phosphatase (AP) under a CMV promoter was used as a reporter gene. RESULTS: The resulting complex was completely insensitive to serum inactivation. Tail vein injection of a 80 micrograms DNA into Balb C mice yielded significant levels of reporter enzyme activity in the lung, heart, spleen, muscle, and liver. Less AP activity was observed in the kidney. No AP activity was observed in blood, bone marrow or brain. A titration of the lipid (DOSPA) to DNA-nucleotide ratio showed the optimal molar ratio for in vivo gene transfer to be 1/1. Using this ratio in a dose response study showed approximately 80 micrograms of DNA/mouse yielded the highest level of gene expression. Using this dose at a 1/1 lipid to DNA nucleotide ratio, the time course for alkaline phosphatase activity was determined. Maximal AP activity was observed 24 hours after injection for all tissues. By day 5, the activity dropped approximately 10 fold for all tissues. By day 7, residual activity was detected in the lung, heart, and muscle. Histology of the lung showed both interstitial and endothelial cells to be transfected. In all other tissues, however, endothelial cells were the only transfected cell type. CONCLUSIONS: These results demonstrate that reformulation of an existing cationic lipid can result in the formation of a stable lipid/DNA complex, which is able to reproducibly transfect lung, heart, spleen, and liver upon intravenous administration. PMID- 9210192 TI - The use of sonication for the efficient delivery of plasmid DNA into cells. AB - PURPOSE: Ultrasonic methods have considerable potential for the introduction of macromolecules into cells. In this paper we demonstrate that, under controlled conditions, application of 20 kHz ultrasound to a suspension of yeast cells facilitates the delivery of plasmid DNA into these cells. METHODS: Aliquots of growing yeast cells (Saccharomyces cerevisae, strain AH22) were suspended in buffer and exposed to 20 kHz ultrasound from a laboratory (probe-type) sonicator in the presence of microgram quantities of plasmid DNA. Efficiency of DNA delivery was scored as the number of cells transformed. RESULTS: Cell transformation was optimal at 30 seconds sonication using an output of 2.0 watts and resulted in a 20 fold enhancement over control values. At extended sonication times, fewer cells showed evidence of transformation because of reduced cell viability. The increased DNA uptake and the decreased cell viability were both attributable to acoustic cavitation events during sonication. The extent of acoustic cavitation was measured and it was found that there was an increase in cavitation events with increased sonication time. Cell viability was shown to be directly related to the number of cavitation events. The effects of sonication on plasmid DNA were investigated and indicated that the structural integrity of plasmid DNA was unaffected by the sonication conditions employed. CONCLUSIONS: Under controlled conditions, ultrasound is an effective means of delivering plasmid DNA into cells. The subsequent expression of DNA molecules in cells depends upon a balance between transient cell damage and cell death. PMID- 9210193 TI - Evidence for diminished functional expression of intestinal transporters in Caco 2 cell monolayers at high passages. AB - PURPOSE: To investigate the effects of passaging on the intrinsic membrane transport parameters of compounds absorbed by means of passive and carrier mediated processes in the Caco-2 cell line. METHODS: Caco-2 cells at low (28-36) and high (93-108) passage numbers were used to evaluate the transport characteristics of model compounds for paracellular diffusion (mannitol), transcellular diffusion (progesterone) and carrier-mediated transport (cephalexin, cephradine, phenylalanine, proline, and taurocholic acid) using side by-side diffusion chambers. Intrinsic intestinal transport parameters were determined by correcting the effective permeability for potential biases introduced by the microporous filter and aqueous boundary layer. Intrinsic maximal flux (Jmax), Michaelis constant (K(m)) and carrier permeability (Pc) were determined as a function of passage number. RESULTS: Compared to the low passaged cells, the high passaged Caco-2 cells were characterized by less morphological heterogeneity, higher transepithelial electrical resistance, higher transcellular diffusion, lower paracellular diffusion, lower carrier-mediated transport and lower alkaline phosphatase activity. The use of effective transport parameters overestimated the K(m) and underestimated Pc but had no effect on Jmax. CONCLUSIONS: The current results provide experimental evidence that the passaging process significantly affects the biological characteristics and transport properties of Caco-2 cell monolayers. The effects are consistent with a reduction in the functional expression of a brush border enzyme and several transport proteins as passage number is increased. The underlying basis for this appears to be a selection of fast-growing subpopulations from the original heterogeneous Caco-2 cell line during passaging. PMID- 9210194 TI - In vitro permeability across Caco-2 cells (colonic) can predict in vivo (small intestinal) absorption in man--fact or myth. AB - PURPOSE: To evaluate and optimize the use of Caco-2 cell monolayers to predict the in vivo absorption of a broad range of compounds in man. METHODS: Caco-2 cells are derived from human adenocarcinoma colon cells and spontaneously differentiate when grown on porous polyethylene terephthalate membranes (PETP) in a 12 well format to form monolayers of polarized cells possessing function similar to intestinal enterocytes. Transport experiments were conducted using 21 day cultured cells in a shaking water bath at 37 degrees C. Radiolabeled mannitol was used to determine monolayer integrity. Apparent permeability coefficient (Papp) was calculated from the appearance of drug in the receiver side. RESULTS: A strong correlation was observed between in vivo human absorption and in vitro Papp for a variety of compounds (R = 0.95, N = 35). For compounds that are substrates of p-glycoprotein (Pgp), use of a Pgp inhibitor resulted in a better estimate of absorption in humans. The results of this study suggest that the overall ranking of compounds with Papp < 1 x 10(-6) cm/sec, between 1-10 x 10(-5) cm/ sec, and > 10 x 10(-6) cm/sec can be classified as poorly (0-20%), moderately (20-70%) and well (70-100%) absorbed compounds, respectively. CONCLUSIONS: These data suggest that Caco-2 cells developed under the culturing and transport conditions defined herein can be used to predict in vivo human absorption of compounds regardless of transport mechanism, viz., transcellular, paracellular and carrier-mediated. PMID- 9210196 TI - Membrane transport in hepatic clearance of drugs. I: Extended hepatic clearance models incorporating concentration-dependent transport and elimination processes. AB - PURPOSE: The objective of the present study was to develop hepatic clearance models which incorporate a unidirectional carrier-mediated uptake and bidirectional diffusional transport processes for drug transport in the sinusoidal membrane of hepatocytes as well as nonlinear intrinsic elimination. METHODS: Two models were derived which view the liver as two separate compartments, i.e., sinusoid and hepatocyte. Model I assumes the instantaneous complete mixing of drugs within each compartment (similar to that of the "well stirred" model), while model II assumes that the drug concentrations in both compartments decrease progressively in the direction of the hepatic blood flow path (similar to that of the "parallel-tube" model). Computer simulations were performed using a range of steady-state infusion rates for a substrate, while varying the Vmax (capacity) and Km (Michaelis-Menten constant) for the carrier mediated uptake process, the diffusional clearance, the Vmax and Km for the intrinsic elimination process, blood flow and protein binding. RESULTS: Simulations in which Vmax and Km for the sinusoidal membrane transporter and the diffusional clearance were varied, demonstrated that these membrane transport processes could affect the clearance of drugs to a significant extent in both models. The estimates for clearance of substrates with the same pharmacokinetic parameters are always lower in model I than in model II, although the quantitative differences in parameter estimates between models varied, depending on the steady state infusion rates. CONCLUSIONS: These more general hepatic clearance models will be most useful for describing the hepatic clearance of hydrophilic compounds, such as organic anions or cations, which exhibit facilitated uptake and limited membrane diffusion in hepatocytes. PMID- 9210195 TI - Influence of morphometric factors on quantitation of paracellular permeability of intestinal epithelia in vitro. AB - PURPOSE: The relative contribution of the small and large intestine to paracellular absorption is a subject of some controversy. Direct comparison of paracellular permeability in different epithelia is complicated by variations in junctional density and/or the absorptive surface area. METHODS: This study used a combination of morphometric analyses and in vitro absorption studies to define permeability characteristics in relation to the amount of paracellular pathway present in rat ileum, colon and the model epithelium, Caco-2. RESULTS: Mucosal to serosal amplification was higher in ileum (3.9) than colon (1.9) or Caco-2 (1). Tight junctional density (lp) of ileal crypts was approximately 3 fold greater (91 m/cm2) than that measured in ileal villi, colonic surface and crypt cells or Caco-2 monolayers (34-37 m/cm2). However, when the relative contributions of the crypts and villi was taken into account there was no significant difference in the mean lp per mucosal area for the three epithelia studied. Using these data to correct for morphometric differences the permeabilities of a range of small hydrophilic molecules (atenolol, D-PheAsp and PEG oligomers MW 282-634) was measured. Permeability of rat ileum and colon were virtually identical for all compounds studied. In contrast, Caco-2 monolayers showed a significantly lower permeability than intestinal tissues with the difference increasing markedly with molecular size. CONCLUSIONS: These studies suggest the importance of accounting for morphological variation when comparing the permeability characteristics of different epithelial systems. PMID- 9210197 TI - Membrane transport in hepatic clearance of drugs. II: Zonal distribution patterns of concentration-dependent transport and elimination processes. AB - PURPOSE: The objective of the present simulation study was to investigate the effects of hepatic zonal heterogeneity of membrane transporter proteins and intrinsic elimination activities on hepatic clearance (CL) and drug concentration gradient profiles in the sinusoidal blood and hepatocytes. METHODS: The model used in the simulations assumes an apparent unidirectional carrier-mediated transport and a bidirectional diffusion of substrates in the hepatic sinusoidal membrane as well as a nonlinear intrinsic elimination. Three different distribution patterns of the transporter and the metabolizing enzyme along the sinusoidal flow path were used for the simulations. The effects of changes in the Michaelis-Menten parameters for those nonlinear processes, and in the unbound fractions of the drug in blood and tissue components were investigated. RESULTS: Significant differences in CL occurred when the distribution patterns of the transporter and/or the metabolizing enzyme activities were altered under nonlinear conditions. The highest CL values were observed when the transporter and the metabolizing enzyme had similar distribution patterns within the liver acinus, while opposite distribution patterns produced the lowest CL values. Tissue concentration profiles were significantly affected by the distribution patterns of the transporter, but the changes in blood concentration profiles were relatively small. Altering protein binding in blood produced significant changes in CL, and blood and tissue concentration gradients, while altering protein binding in tissue affected only drug accumulation patterns within hepatocytes, regardless of the distribution patterns of the transporter or the metabolizing enzyme. CONCLUSIONS: The present simulations demonstrate that hepatic zonal heterogeneities in the transporter and the metabolizing enzyme activities can significantly influence hepatic clearance and/or drug concentration gradient profiles in the sinusoidal blood and hepatocytes. PMID- 9210198 TI - Role of brain tissue localized purine metabolizing enzymes in the central nervous system delivery of anti-HIV agents 2'-beta-fluoro-2',3'-dideoxyinosine and 2' beta-fluoro-2',3'-dideoxyadenosine in rats. AB - PURPOSE: This study examines the central nervous system (CNS) delivery of 2'-beta fluoro-2',3'-dideoxyadenosine (F-ddA) and 2'-beta-fluoro-2',3'-dideoxyinosine (F ddI), acid stable analogues of dideoxyadenosine (ddA) and dideoxyinosine (ddI) having reduced susceptibility to purine salvage pathway enzymes important in the metabolism of ddA and ddI, adenosine deaminase (ADA) and purine nucleoside phosphorylase (PNP), respectively. Their CNS delivery compared to that for ddI provides insight into the role of brain tissue ADA and PNP in these processes. METHODS: Brain and cerebrospinal fluid (CSF) concentration-time profiles were obtained for F-ddI during and after intravenous infusions of F-ddI, and for both F-ddA and F-ddI after F-ddA infusions in normal rats or rats pre-treated with the ADA inhibitor 2'-deoxycoformycin (DCF). Rate constants for CNS entry, efflux and metabolism were estimated by computer fits using plasma concentration-time profiles as the driving force functions. RESULTS: The CNS delivery of F-ddI did not differ significantly from that for ddI. F-ddA, which is more lipophilic than F-ddI, provided higher brain (approximately 8x) and CSF (approximately 11x) concentrations of total dideoxynucleoside (F-ddA and F-ddI) compared to F-ddI. Deamination by brain tissue ADA to form F-ddI reduced CNS levels of intact F-ddA but provided higher brain parenchyma (5x) and CSF/plasma (3x) ratios of F-ddI relative to F-ddI controls. Thus, F-ddA functions in part as a CNS-activated prodrug of F-ddI. DCF pre-treatment inhibited brain tissue ADA, abolishing the prodrug effect, and enhancing F-ddA concentrations in both brain parenchyma (5x) and CSF (6x). CONCLUSIONS: PNP metabolism does not appear to play a role in the low CNS delivery of ddI. On the other hand, deamination of F-ddA by brain tissue ADA is an important process, such that F-ddA functions in part as a CNS-activated prodrug of F-ddI. Enhanced CNS uptake of intact F-ddA can be achieved with ADA inhibition. PMID- 9210199 TI - Prednicarbate biotransformation in human foreskin keratinocytes and fibroblasts. AB - PURPOSE: Evaluation of skin layer-specific prednicarbate (PC) biotransformation, possibly explaining the improved benefit/risk ratio of this topical corticosteroid in atopic dermatitis (1,2). METHODS: Metabolism of PC in keratinocyte and fibroblast monolayers derived from human juvenile foreskin was evaluated. Drug concentration was determined by HPLC/UV-absorption. Accompanying cell viability tests (MTT-tests) were performed to exclude toxic drug effects. RESULTS: Keratinocytes hydrolyzed the double ester PC (2.5 x 10(-5) M) at position 21 to the monoester prednisolone 17-ethylcarbonate (P17EC) which nonenzymatically transformed to prednisolone 21-ethylcarbonate (P21EC). This metabolite was enzymatically cleaved to prednisolone (PD), the main biotransformation product at 24 hours. Fibroblasts, however, showed a distinctively lower enzyme activity. Both, PC and P17EC (or rather P21EC) were hydrolyzed to a minor extent only. The biotransformation pathway, however, was the same. When P17EC was added separately, it transformed to P21EC and again was cleaved by keratinocytes to a much higher extent. Despite of the rather high glucocorticoid concentration MTT-tests proved a non-disturbed cell viability and proliferation rate. CONCLUSIONS: Extrapolating our results to the in-vivo situation, topically applied PC may be metabolized by epidermal cells during skin penetration. A complex mixture of compounds reaches the dermis, whose fibroblasts are barely able to metabolize the steroids. Since skin atrophy is less pronounced with PC as compared to conventional halogenated glucocorticoids, less potent PC metabolites appear to be the dominant species in the dermis. PMID- 9210200 TI - Evaluation of amorphous ursodeoxycholic acid by thermal methods. AB - PURPOSE: The purpose of this study was to characterize the amorphous state of ursodeoxycholic acid (UDCA) samples by using isothermal microcalorimetry, X-ray diffraction, infrared (IR) spectroscopy and solid state carbon 13 nuclear magnetic resonance (13C-NMR) spectroscopy, and to demonstrate the application of the thermal methods (microcalorimetry and differential scanning calorimetry (DSC) for studying the amorphous state and clarifying the dissolution mechanism of UDCA. METHODS: Amorphous UDCA was prepared by grinding and rapid cooling of the melts. The heat of solution of UDCA was measured by an isothermal heat-conduction twin microcalorimeter at 25.0 degrees C. Some physicochemical properties of amorphous UDCA were also studied. RESULTS: The intensities of X-ray diffraction peaks of crystalline UDCA decreased with an increase in grinding time. The heat levels of solution of crystalline UDCA and UDCA ground for 1 min were endothermic, and became exothermic with an increase in grinding time. A good correlation was obtained between the heat of solution and the heat of crystallization determined from the peak area in DSC. Although no significant difference was observed in X-ray diffraction patterns of amorphous UDCA prepared by the two methods, significant differences were recognized in DSC, IR and 13C NMR, and the heat of solution indicated different values among the two samples. The stability of amorphous UDCA samples stored under 74.5% relative humidity at 40 degrees C was found to depend upon the preparation methods. CONCLUSIONS: Different states of amorphous UDCA were obtained depending on the preparation method. The application of thermal methods to evaluate the amorphous state was demonstrated. The mechanism of dissolution of UDCA was discussed from the results of the heat of solution examination. PMID- 9210201 TI - The crystallite-gel-model for microcrystalline cellulose in wet-granulation, extrusion, and spheronization. AB - PURPOSE: A new model for the wet-extrusion/spheronization process with microcrystalline cellulose (MCC) is proposed. The crystallite-gel-model is able to elucidate the unique role of MCC in this process. Many other experimental results, which cannot be explained by the standard model of granulation, the liquid saturation model, give evidence for the crystallite-gel-model. METHODS: Pellets were prepared from different types of MCC. Water content during extrusion, power consumption and aspect ratio were correlated. X-ray diffractograms of MCC powders, extrudates and pellets were taken in order to provide information on changes at the single crystallite level. SEM-photographs and leaching studies gave additional information on changes at the particulate level of MCC. RESULTS: At the level of MCC powder particles, dramatic changes occurred during extrusion/spheronization. In contrast to this no changes could be observed at the level of individual crystallites. CONCLUSIONS: During granulation and extrusion MCC-particles are thought to be broken down into smaller particles and possibly ultimate single crystallites in the presence of water. The crystallite-gel-model serves as the framework for a new interpretation of the wet extrusion/spheronization process. Apart from the ability to explain experimental data published previously in the literature it can be used to develop new experimental plans for further research. Consequently, the crystallite-gel-model exhibits explanatory as well as predictive power. PMID- 9210202 TI - Radiation sterilization of formoterol. AB - PURPOSE: Radiation sterilization is becoming increasingly popular for the sterilization of many pharmaceutical products. We have investigated the gamma radiation induced effects on formoterol fumarate by HPLC and ESR spectroscopy. RESULTS AND DISCUSSION: Numerical simulation of the evolution of the ESR signal versus dose was performed using linear regression, quadratic fit and power function. The shape of the dosimetric curve is linear in the range 5-30 kGy. Owing to the weak number of free radicals generated during the irradiation, the accuracy of measurements is low. For a dose of 25 kGy, discriminating irradiated from unirradiated samples is possible if the storage period is less than 250 days. The comparison between chromatographic profiles of irradiated and unirradiated samples showed minor differences. CONCLUSIONS: From our preliminary results, radiosterilization of formoterol fumarate may be technically feasible. Estimation of the irradiation dose by ESR may be possible but, due to the weak number of free radicals generated during the irradiation, the accuracy of measurements appeared low. PMID- 9210203 TI - Development of a novel controlled-release system for gastric retention. AB - PURPOSE: We report on the development of a novel controlled-release gastric retention system, which consists of a matrix tablet, coated with a permeable membrane. When immersed in simulated gastric fluid, the tablet expands. The tablet remains expanded for eighteen to twenty hours, during which time the drug is released. The tablet then either disintegrates into fragments or loses its integrity. METHODS: Tablets containing a soluble drug (chlorpheniramine maleate, i.e., CPM) and a poorly soluble drug (riboflavin 5' phosphate, i.e., R5'P) were compressed. They were coated with a permeable and elastic polymer (Eudragit). Dissolution profiles of these tablets were studied. The changes in the pH, viscosity, and deformation characteristics as a function of time were measured. RESULTS: Carbopol provided a firm structure to the swollen tablet. Polyvinyl pyrrolidone XL (PVP XL) contributed to the swelling of the tablet. Carbonates provided the initial alkaline micro-environment for Carbopol to gel and conferred buoyancy to the tablet. Coating provided the support needed for the core to remain intact during drug release and, at the same time, it allowed drug release due to its permeable nature. During release, the gelling properties of Carbopol lessened, resulting in a decrease in the firmness of the core. This was evident from the decrease in the viscosity of the core. The energy required at 50% strain also decreased as the drug release progressed. CONCLUSIONS: When this tablet is ingested, the chances of its elimination through the pylorus should be greatly reduced due to tablet's expansion, and due to its disintegration or loss in integrity it should then be expelled out of the stomach at the end of the drug release. PMID- 9210204 TI - In vitro/in vivo comparison of drug release and polymer erosion from biodegradable P(FAD-SA) polyanhydrides--a noninvasive approach by the combined use of electron paramagnetic resonance spectroscopy and nuclear magnetic resonance imaging. AB - PURPOSE: The purpose of this study was to compare drug release and polymer erosion from biodegradable P(FAD-SA) polyanhydrides in vitro and in vivo in real time and with minimal disturbance of the investigated system. METHODS: P(FAD-SA) 20:80 and P(FAD-SA) 50:50 polymer tablets were loaded with the spin probe 3 carboxy-2,2,5,5-tetramethyl-pyrrollidine-1-oxyl (PCA) and implanted subcutaneously in the neck of rats or placed in 0.1 M phosphate buffer. 1.1 GHz EPR spectroscopy experiments and 7T MRI studies (T1 and T2 weighted) were performed. RESULTS: A front of water penetration was visible by MRI in vitro in the case of P(FAD-SA) 20:80, but not for P(FAD-SA) 50:50. For both polymers, the thickness of the tablets decreased with time and a insoluble, easy deformable residue remained. Important processes such as edema, deformation of the implant, encapsulation and bioresorption were observable by MRI in vivo. P(FAD-SA) 50:50 was almost entirely absorbed by day 44, whereas an encapsulated residue was found for P(FAD-SA) 20:80 after 65 days. The EPR studies gave direct evidence of a water penetration induced changes of the microenvironment inside the tablet. EPR signals were still detectable in P(FAD-SA) 20:80 implants after 65 days, while the nitroxide was released in vitro within 16 days. CONCLUSIONS: Important parameters and processes such as edema, deformation of the tablet, microviscosity inside the tablet and encapsulation can be monitored in real time by the combined use of the noninvasive techniques MRI and EPR leading to better understanding of the differences between the in vitro and in vivo situation. PMID- 9210206 TI - Isolation and characterization of an acetylated impurity in Escherichia coli derived recombinant human interleukin-10 (IL-10) drug substance. PMID- 9210207 TI - Neuroendocrine responses to fenfluramine challenge are influenced by exposure to chronic social stress in adult male cynomolgus macaques. AB - This study was designed to investigate the effect of chronic social stress on central serotonergic responsivity in adult male cynomolgus monkeys (Macaca fascicularis). The influences of social stress and dominance status (social rank) on adrenocorticotropin hormone (ACTH) and cortisol responses to acute administration of an indirect serotonergic agonist (fenfluramine) were evaluated in 75 cynomolgus macaques that were housed in five-member social groups for 28 mo. These groups either remained stable in composition (No-Stress) or had their composition periodically reorganized in the first (Early-Stress) or second (Late Stress) halves of the study. At the end of the 23rd month, a fenfluramine challenge was done. Animals in the Late-Stress condition had significantly higher ACTH responses compared to those in the No-Stress condition (p < .05) and significantly higher cortisol responses compared to those in the Early-Stress condition (p < .05). No differences between dominant and subordinate animals in ACTH or cortisol responses to challenge were identified. These data suggest that social stress produces a "state"-related augmentation of hypothalamic-pituitary adrenal responsivity to fenfluramine (serotonergic) challenge in cynomolgus macaques. PMID- 9210205 TI - Free fatty acids cause pH-dependent changes in drug-lipid membrane interactions around physiological pH. AB - PURPOSE: The influence of oleic acid (OA) and phosphatidylethanolamine (PhE) as membrane constituents on the partition behavior of (RS)-[3H]propranolol between unilamellar liposomes and buffer was studied as a function of pH. METHODS: Partition studies were performed by means of equilibrium dialysis at 37 degrees C over a broad pH range at a molar propranolol to lipid ratio in the membrane of 10(-6). RESULTS: As compared to the standard phosphatidylcholine (PhC) liposome/buffer partition system PhE and OA have an enhancing effect on the apparent partition coefficient (D) of (RS)-[3H]propranolol between pH 6 and 11. Data analysis with Henderson-Hasselbalch equations revealed that the neutral propranolol has a higher affinity to membranes containing net neutral charged PhE than to pure PhC-liposomes. Net negatively charged PhE seems to have no significant influence on the partitioning. Deprotonated OA caused an increase in the true partition coefficient (P) of the protonated propranolol. Neutral OA showed no influence on the partitioning. From the fit D vs pH curves and from zeta potential measurements of the liposomes the intrinsic pKa values of the membranous lipids were calculated as 7.5 to 7.8 for OA and 9.7 to 9.8 for PhE. CONCLUSIONS: Since the pKa of membranous OA is close to the physiological pH and D depends on the ionisation state of OA, small pH changes around the physiological pH may cause large differences in drug-lipid membrane interactions. PMID- 9210208 TI - The stability of plasma growth hormone and MHPG responses to repeated clonidine challenge in normal males. AB - Clonidine is a centrally acting alpha 2-adrenergic agonist used in many psychiatric studies to assess adrenergic functioning. The short- and long-term stability of plasma growth hormone (GH) and plasma 3-methoxy-4-hydroxy phenylglycol (MHPG) responses to clonidine (2 micrograms/kg IV) over a 60-min period were assessed in subsets of 13 male normal controls on 2 consecutive days (Study A; n = 11) and on 2 days separated by several months (Study B; n = 11). In Study A, no significant differences between consecutive days were found in either baseline plasma GH or MHPG or their responses to clonidine. The 60 minute plasma GH responses between consecutive days were highly correlated (r = 0.75, n = II, p < .001), while the 60 min plasma MHPG responses were not. In Study B, no significant differences in baseline plasma GH or MHPG, or their responses to clonidine challenge, were found between the 2 test days. However, neither the plasma GH responses nor the plasma MHPG responses to clonidine at 60 min correlated significantly between the 2 study days separated by several months. Both in Study-A and in Study B, 8 of 11 subjects had a stable GH response to clonidine across both study days when defined dichotomously (blunted < 4 ng/ml; otherwise, not blunted). These results suggest that the plasma GH response and plasma MHPG response to clonidine are unaffected by repeat clonidine challenge separated by 24 h, and that the plasma GH response to clonidine may be more stable over time than the plasma MHPG response to clonidine. PMID- 9210209 TI - Effects of menstrual cycle on creativity. AB - Seventeen healthy women (21-31 years) not taking oral contraceptives were tested at three phases of the menstrual cycle distinctly differing in hormonal patterns (menses, preovulatory phase, and midluteal phase). Phase detection was assured by determination of blood hormone concentrations. Another 17 women taking oral contraceptives (combined preparation of estrogen and progestin) served as age matched controls, and were tested during menses and during phases corresponding to preovulatory and midluteal phase. On each test occasion, aspects of creativity were assessed by a battery of six tests measuring "semantic" and "figural" abilities of divergent thinking. Additionally, a test of motor perseveration (Mittenecker-Zeigeversuch) was presented. During the preovulatory phase, creativity was in general improved when serum concentrations of estrogen (E2) and luteinizing hormone (LH) were highest whereas motor perseveration decreased. In control women, there was no preovulatory improvement of divergent thinking and no preovulatory decrease in motor perseveration. PMID- 9210210 TI - The influence of aging on the plasma concentration and renal clearance of homovanillic acid. AB - Using percutaneously placed arterial and venous catheters, we examined the influence of aging on the plasma concentration, whole body production rate, and renal clearance of homovanillic acid (HVA) in 60 healthy adult volunteers. The arterio-renal fractional extraction of HVA combined with the renal plasma flow (Fick Principle) were used to estimate the whole body HVA production rate and the renal plasma HVA clearance. The arterial HVA plasma concentration, whole body rate of HVA production, and HVA plasma clearance were determined to be 54 +/- 3 nmol/l, 27 +/- 2 nmol/min and 502 +/- 32 ml/min, respectively. The resting arterial HVA plasma concentration was positively correlated with aging, with the increased HVA concentrations in the older subjects being due to a diminished renal HVA plasma clearance. The diminished clearance of HVA occurred in response to a decreased renal plasma flow; the fractional extraction of HVA across the kidney remained unchanged with aging. This study emphasises the need for using age-matched control groups in neurochemical and neuropsychiatric studies, and demonstrates that increases in the arterial level of HVA do not necessarily reflect an increased rate of HVA production but may arise due to a diminished excretion rate of HVA from the body. PMID- 9210211 TI - Stress-induced regulation of the renal peripheral benzodiazepine receptor: possible role of the renin-angiotensin system. AB - The etiology of the decrease in renal peripheral benzodiazepine receptor (PBR) binding caused by stress was studied in rats. Prior investigations suggest that the response of the renal PBR to stress occurs independently of the hypothalamo pituitary-adrenal (HPA) axis and sympathetic nervous system. The present experiments tested the hypothesis that the renin-angiotensin system is involved in regulating the PBR. Eighty min of brief, intermittent tailshocks caused increases in plasma renin activity and decreases in renal PBR binding. The stress induced decrease in renal PBR binding was reversed by pretreatment with captopril. Acute administration of angiotensin II (ANG II) alone caused reductions in PBR binding in kidney, heart, and cerebral cortex. These data suggest that ANG II may be an endogenous factor responsible for regulating the PBR in several tissues during stress. PMID- 9210213 TI - Peripheral-type benzodiazepine receptors on human blood mononuclear cells are not regulated by ovarian steroids. AB - Peripheral-type benzodiazepine receptors (pBZr) have been shown to be sensitive to hormonal perturbations, including changes in ovarian steroids. This study examines whether estradiol and progesterone modulate pBZr binding in membranes of human blood mononuclear cells, as measured by binding of both 3H-PK 11195 and 3H Ro 5-4864. Our findings were negative. There was no steroidal modulation of pBZr binding to these membrane preparations in vivo in normal women studied at different sex-steroid phases of the menstrual cycle, or during 8-30 weeks of pregnancy. There was also no effect of hormones on the binding sites in cultures of mononuclear cells treated with estradiol or progesterone (up to 10(-5) M) over a period between 2 and 72 h. Further, we performed in vitro competition experiments, which showed that both steroids are not active at the pBZr. Our data suggest that pBZr located in human blood mononuclear cells are insensitive to the physiological variations of circulating female hormones. PMID- 9210212 TI - Cognitive and neuroradiological findings in congenital adrenal hyperplasia. AB - Nineteen patients with congenital adrenal hyperplasia (CAH) aged over 16 years were given a neuropsychological evaluation; no significant differences with individually matched normal controls were detected. CAH subjects, however, revealed slightly higher IQs with respect to the expected distribution. No significant learning disabilities could be detected. Fifteen patients underwent nuclear magnetic resonance (NMR); 4 subjects showed small areas of increased signal intensity in the white matter, without prevalence of side; this finding did not correlate with clinical and cognitive characteristics. The results are discussed in the light of possible hormonal influences. PMID- 9210214 TI - Effects of age on spontaneous cortisolaemia of normal volunteers and depressed patients. AB - The aim of this study was to characterize the relationships between age and endogenous cortisol secretion in healthy controls and in major depressed patients between 18 and 58 years of age. Toward this end, the authors measured morning basal plasma cortisol secretion every 30 min from 0900 h until 1100 h and computed the integrated morning cortisol secretion in 80 normal controls and 118 major depressed patients. A significant negative correlation between age and plasma morning cortisol was found in normal volunteers but not in major depressives. The observed decrease in cortisol secretion with age in normal controls older than 35 years does not occur in major depressives. The middle age (+/- 35 years) appears to be an important turning point in hypothalamo-pituitary adrenal (HPA)-axis function of normal persons vs. major depressives. PMID- 9210215 TI - Role of oxytocin in the neuroadaptation to drugs of abuse. AB - Oxytocin (OXT), a neurohypophyseal hormone, has a wide range of behavioral effects outside its classic peripheral endocrine functions. OXT involvement in adaptive central nervous system processes has been demonstrated as an inhibitory, amnestic action on learning and memory in different paradigms. Because adaptation and learning are likely to be involved in the neural events leading to drug tolerance and dependence, the question logically arose whether OXT is able to influence the development of tolerance of and dependence on abused drugs. In this review, we summarize our results on the effects of OXT on opiate (including morphine, heroin, and the endogenous opiates beta-endorphin and enkephalin) tolerance and dependence, heroin self-administration, psychostimulant-induced behavioral changes, and behavioral tolerance and sensitization. The sites and mechanisms of action and the possible physiological role of OXT are also discussed. In the first part of this review the effects of exogenously administered OXT on both the acute and chronic behavioral effects of opiates and psychostimulants have been summarized. OXT inhibited the development of tolerance to morphine, heroin, beta-endorphin, and enkephalin, OXT also inhibited the development of cross-tolerance between the predominantly mu-agonist heroin and the predominantly delta-agonist enkephalin in mice. Naloxone-precipitated morphine withdrawal syndrome was also attenuated by OXT. Heroin self administration was decreased by OXT administration in heroin-tolerant rats. OXT inhibited cocaine-induced exploratory activity, locomotor hyperactivity, and stereotyped behavior in rats and in mice. Behavioral tolerance to cocaine was also attenuated by OXT. On the contrary, OXT stimulated the development of behavioral sensitization to cocaine. OXT did not alter the stereotyped behavior induced by amphetamine. In the second series of experiments, the sites of action of OXT on drug-related behavior were investigated. Intracerebro-ventricular (ICV) and intracerebral (IC) administration of an OXT-receptor antagonist inhibited the effects of peripherally administered OXT on morphine tolerance, heroin self administration, and cocaine-induced sniffing behavior. This suggests the central, intracerebral location of OXT target sites. Local IC microinjection of OXT in physiological doses into the posterior olfactory nucleus, tuberculum olfactorium, nucleus accumbens, central amygdaloid nucleus, and the hippocampus inhibited the development of tolerance to and dependence on morphine as well as cocaine-induced sniffing behavior and tolerance to cocaine. The physiological role of endogenous OXT in acute morphine tolerance has also been demonstrated, since OXT antiserum (ICV) and OXT-receptor antagonist (injected into the basal forebrain structures) potentiated the development of morphine tolerance. Finally, we investigated the possible mechanisms of action of OXT on drug related behavior. Both morphine tolerance and dependence, and cocaine administration, increased dopamine utilization in the mesencephalon and in the nucleus accumbens, respectively. OXT treatment decreased the alpha-methylparatyrosine-induced dopamine utilization in the mesencephalon and in the nucleus accumbens-septal complex. Chronic OXT treatment decreased the number of apparent binding sites of dopamine in the basal forebrain area. It also inhibited a cocaine-induced increase in dopamine utilization in the nucleus accumbens, but not in the striatum. In light of this information, it appears that OXT inhibits the development of opiate tolerance, dependence, and self-administration as well as the acute behavioral actions of and chronic tolerance to cocaine. This suggests the possible role of this neuropeptide in the regulation of drug abuse. Therefore, OXT may act as a neuromodulator on dopaminergic neurotransmission in limbic-basal forebrain structures to regulate adaptive CNS processes leading to drug addiction. PMID- 9210216 TI - Actin-binding membrane proteins identified by F-actin blot overlays. AB - Actin and associated proteins at the cytoskeleton-plasma membrane interface stabilize the membrane bilayer, control cell shape, and delimit specialized membrane domains. To identify membrane proteins that bind directly to F-actin, we have developed a blot overlay assay with 125I-labeled F-actin. In the soil amoebae, Dictyostelium discoideum, the major proteins reactive in this assay are p30a, a 34-kD peripheral membrane protein that is concentrated in filopodia and at sites of cell-cell adhesion, and ponticulin, a 17-kD transmembrane glycoprotein required for efficient chemotaxis and for control of pseudopod dynamics. Proteins with apparent molecular masses of approximately 34- and approximately 17-kD also are observed on F-actin blot overlays of many mammalian cell lines. However, in mammalian cells, the most prominent F-actin binding proteins in this assay exhibit apparent molecular masses of 78-, 80-, 81-, approximately 120-, and 205-kD. Bovine neutrophils contain the 78-, 81-, and 205 kD proteins, all of which co-isolate with a plasma membrane-enriched fraction. We have previously identified the 78-, 80-, and 81-kD proteins as moesin, radixin, and ezrin, respectively. These proteins, which are members of the protein 4.1 superfamily, colocalize with actin in cell surface extensions and have been implicated in the protrusion of microvilli, filopodia, and membrane ruffles. The 205-kD protein (p205) appears to be absent from current databases, and its characteristics are still under investigation. We here report that the 120-kD protein is drebrin, a submembranous actin-binding protein originally identified as a developmentally regulated brain protein. Thus, it appears that F-actin blot overlays provide an efficient assay for simultaneous monitoring of a subset of F actin binding proteins, including p30a, ponticulin, moesin, radixin, ezrin, p205, and drebrin. PMID- 9210217 TI - Interactions between dystrophin and the sarcolemma membrane. AB - Dystrophin serves as a link between the subsarcolemmal cytoskeleton and the extracellular matrix. The NH2 terminus attaches to the cytoskeleton, while the COOH terminus attaches to the dystrophin associated protein (DAP) complex, which can be separated into the dystroglycan, sarcoglycan, and syntrophin subcomplexes. While the function of each DAP is not known, the dystroglycan complex binds laminin in the extracellular matrix, and binds the dystrophin COOH terminus in vitro. The syntrophins also bind the dystrophin COOH terminus in vitro, but no evidence has been reported for an interaction between dystrophin and the sarcoglycans. Human mutations have been found in dystrophin, the sarcoglycans and laminin, all of which lead to various types of muscular dystrophy. We have been studying the dystrophin domains necessary for formation of a functional complex by generating transgenic mdx (dystrophin minus) mice expressing internally truncated dystrophins. These mice provide in vivo models to study the localization of truncated dystrophin isoforms, the association of the truncated proteins with the DAP complex, and the functional capacity of the assembled DAP complexes. Expression of a dystrophin deleted for most of the NH2-terminal domain in mdx mice leads to only a mild dystrophy, indicating that dystrophin can attach to the cytoskeleton by multiple mechanisms. Truncation of the central rod domain leads to normal DAP complex formation and almost fully prevents development of dystrophy. Deletion analysis of the COOH-terminal regions indicates that a broad cysteine-rich domain is indispensable for dystrophin function. This region coincides with the in vitro identified beta-dystroglycan binding domain. Mice lacking this latter domain express very low levels of the sarcoglycans, indicating that the sarcoglycan complex binds dystrophin via dystroglycan. All deletion constructs tested lead to normal expression of the syntrophins, indicating that syntrophin associates with the DAP complex via multiple binding partners. PMID- 9210218 TI - A multiple site, side binding model for the interaction of dystrophin with F actin. PMID- 9210219 TI - Roles of the membrane cytoskeleton in protein sorting. PMID- 9210220 TI - Role of cytoplasmic motors in post-Golgi vesicular traffic. PMID- 9210221 TI - Molecular architecture of tight junctions: occludin and ZO-1. PMID- 9210222 TI - Capping actin filament growth: tropomodulin in muscle and nonmuscle cells. AB - Actin filament lengths are precisely regulated and very stable in the sarcomeres of striated muscle, in the erythrocyte membrane skeleton, and in cell protrusions such as microvilli in intestinal epithelial cells and stereocilia in hair cells of the inner ear. In contrast, in motile cells, actin filament lengths are dynamically regulated when cells extend lamellipodia. Control of actin filament lengths and dynamics in cells is expected to be achieved in part by capping proteins that prevent filament growth or shrinkage by blocking subunit exchange at both the fast growing (barbed) and slow growing (pointed) filament ends. Much is known about how barbed end capping proteins control actin filament assembly and length in many cells, but little is known about the significance of regulating actin filament assembly at the pointed end. Tropomodulin is the only known capping protein for the pointed ends of actin filaments and is a approximately 40-kD protein that is expressed in erythrocytes, striated muscle, lens fiber cells, some regions of the adult brain, as well as sensory neurons and epithelial cells of the inner ear. A related isoform (59% identical at the protein level) is expressed principally in neurons of both embryonic and adult brain. In striated muscle and in erythrocytes, tropomodulin is tightly associated with the actin filament pointed ends where it functions to maintain actin filament length in vivo (for recent reviews, see Fowler, 1996; Gregorio and Fowler, 1996). Unlike proteins that cap actin filament barbed ends, tropomodulin also binds tropomyosin and requires tropomyosin for tight capping of actin filament pointed ends. Mapping of functional domains on tropomodulin shows that the COOH-terminal end of tropomodulin is important for actin filament pointed end capping activity while the NH2-terminal portion of tropomodulin contains the tropomyosin binding domain. Searches of protein and EST databases for tropomodulin-like sequences reveal a number of proteins with homologies to both the tropomyosin binding and the actin filament capping portions of tropomodulin. In particular, we have identified a tropomodulin-like 64-kD protein that is principally expressed in smooth muscle cells. We anticipate that tropomodulin and this 64-kD protein are members of a larger family of tropomyosin and actin binding proteins that are responsible for capping actin filament pointed ends and regulating actin filament lengths in muscle and nonmuscle cells. PMID- 9210223 TI - Functional studies of the membrane skeleton in Drosophila: identification of a positional cue that targets polarized membrane skeleton assembly. PMID- 9210224 TI - Molecular architecture of the specialized axonal membrane at the node of Ranvier. PMID- 9210225 TI - Specificity of desmosomal plaque protein interactions with intermediate filaments: keeping adhesive junctions segregated. PMID- 9210227 TI - Functional studies of the protein 4.1 family of junctional proteins in Drosophila. PMID- 9210226 TI - Intermediate filament linker proteins. PMID- 9210228 TI - Cytoskeletal interactions with glutamate receptors at central synapses. AB - The data presented here are clearly just the beginning of any comprehensive understanding of the set of regulatory and cytoskeletal proteins that interact with membrane receptors in the postsynaptic density. They do, however, indicate that both glutamate channels at central excitatory synapses are involved in complex protein-protein interactions. For example, while NR2A is important for Ca dependent inactivation of NMDA receptors, studies in several systems suggest that the other major NR2 subunit in hippocampal neurons, NR2B, predominates at critical times during synapse formation. In addition, the COOH terminus of NR2B binds to several novel cytoskeletal proteins. These results provide circumstantial evidence that NR2B may play specific roles in function and localization of receptors at excitatory synapses. The possible role of NR2B in early synaptic function gains additional support from functional data suggesting that NMDA receptors have specific roles during development (Komuro and Rakic, 1993; Rabacchi et al., 1992; Yen et al., 1993). The essential role of NR1 and NR2B in development is graphically demonstrated by the neonatal death of transgenic mice lacking either of these two subunits (Forrest et al., 1994; Kutsuwada et al., 1996) whereas NR2A and NR2C-deficient mice are less severely affected (Sakimura et al., 1995; Ebralidze et al., 1996). PMID- 9210229 TI - A membrane skeleton that clusters nicotinic acetylcholine receptors in muscle. AB - We have presented ultrastructural and semiquantitative immunofluorescence evidence to support the idea that AChR are clustered in rat myotubes by virtue of their ability to associate with a spectrin-based membrane skeleton. Many of the interactions postulated to be involved in the formation of this skeleton, and in the anchoring of AChR to it, must still be examined at the biochemical level, but the overall similarity of this structure to that of the human erythrocyte is already clear. The ability of different members of the spectrin superfamily to associate in various combinations to form distinct plasmalemmal domains provides some exciting hints as to how the surface membrane can be organized efficiently to subserve multiple purposes. One of the challenges of future research will be to learn how innervation regulates the assembly of the membrane skeleton at the developing NMJ, and how this structure is altered as the junction matures. Another will be to learn if the principles of neuromuscular synaptogenesis are relevant to interactions between neurons in the brain, where cells must distinguish between multiple synaptic inputs and assemble synaptic structures at thousands of distinct sites on the neurolemma. Members of the spectrin superfamily have been identified in synaptic structures in the central nervous system (e.g., Carlin et al., 1983; LeVine and Sahyoun, 1986; Malchiodi-Albedi et al., 1993), so much of what we have learned at the neuromuscular junction may be applicable to central synapses. PMID- 9210231 TI - Mechanical transduction by ion channels: how forces reach the channel. PMID- 9210230 TI - Syntrophins: modular adapter proteins at the neuromuscular junction and the sarcolemma. PMID- 9210232 TI - Cell signalling and CAM-mediated neurite outgrowth. AB - A wide range of molecules promote nerve growth, and these include cell adhesion molecules (CAMs), NCAM, N-cadherin, and the L1 glycoprotein are CAMs that are normally found on both the advancing growth cone and also on cellular substrates, and in general operate via a homophilic binding mechanism. In recent years it has become clear that nerve growth stimulated by these CAMs does not rely on the adhesion function of these molecules, but instead requires that the CAMs activate second messenger cascades in neurons. A large body of evidence supports the hypothesis that homophilic binding of the CAM in the substrate to the CAM in the neuron leads to activation of the neuronal FGF receptor, possibly via a direct interaction in cis between the CAM and the FGF receptor. The consequential activation of PLC gamma is both necessary and sufficient to account for the neurite outgrowth response stimulated by the above three CAMs. Based on the above model, we reasoned that soluble CAMs might also be able to stimulate neurite outgrowth and that such agents might be developed as potential therapeutic agents for stimulating nerve regeneration. To this end we have made soluble chimeric molecules consisting of the extracellular domain of NCAM or L1 fused to the Fc region of human IgG 1. We have found that these molecules can stimulate neurite outgrowth from rat and mouse cerebellar granule cells cultured on a variety of tissue culture substrates and that they do so by activating the FGF receptor signal transduction cascade in the neurons. Consistent with this model, we find that neurons that have their FGF receptor function ablated as a consequence of the expression of dominant negative FGF receptors, no longer respond to the soluble CAM. Downstream targets of CAM function have also been studied. Addition of soluble CAMs to isolated growth cone preparations from mouse or rat brain leads to enhanced phosphorylation of the GAP-43 protein providing a link between the cell surface and the cytoskeleton. PMID- 9210233 TI - Dissection of protein kinase and phosphatase targeting interactions. AB - Protein phosphorylation is a primary means of mediating signal transduction events that control cellular processes. Accordingly, the activities of protein kinases and phosphatases are highly regulated. One level of regulation is that the subcellular distribution of several kinases and phosphatases is restricted by association with targeting proteins or subunits. This mechanism promotes rapid and preferential modulation of specific targets within a defined microenvironment in response to diffusible second messengers. The type II cAMP-dependent protein kinase (PKA) is targeted by association of its regulatory subunit (RII) with A kinase anchoring proteins (AKAPs). To date, 36 unique AKAPs have been identified. Each of these proteins contains a conserved amphipathic helix responsible for AKAP association with cellular structures. Disruption of PKA/AKAP interaction with peptides patterned after the amphipathic helix region blocks certain cAMP responses, including the modulation of glutamate receptor ion-channel activity in neurons and transcription of cAMP-responsive genes. Yeast two-hybrid screening methods have identified neuronal specific AKAP79-binding proteins including the beta isoform of the phosphatase 2B, calcineurin. Biochemical and immunological studies have confirmed the two-hybrid results and identified additional members of this multienzyme signaling complex, including certain protein kinase C isoforms. These findings are consistent with colocalization of CaN, PKC, and type II PKA by AKAP79 and suggest a novel model for reversible phosphorylation in which the opposing kinase and phosphatase actions are colocalized in a signal transduction complex by association with a common anchor protein. PMID- 9210234 TI - Signaling through the focal adhesion kinase. PMID- 9210236 TI - Philosophical anatomy revisited. PMID- 9210237 TI - Anatomic basis of the transgluteal approach to the hip-joint by anterior hemimyotomy of the gluteus medius. AB - The authors present a study of the intrinsic anatomy of the gluteus medius m, and of its innervation through the caudal branch of the superior gluteal n. The existence of an intramuscular tendon in the thickness of the gluteus medius was constantly prooved in 40 muscles. The relations of the intrinsic fibrous structure of the muscle and its innervation were studied. The authors deduce from that the topography of a gluteus medius incision, with respect to a safety area towards its innervation, which leads to an exposure of the acetabulum that is satisfying and gives opportunities of a sound repair after the surgery of the hip joint through the transgluteal approach. They propose the "anterior hemimyotomy of the gluteus medius m" designation. PMID- 9210235 TI - Involvement of the actin network in insulin signalling. AB - The purpose of the studies included in this chapter was to examine the role of the actin network in the propagation of insulin action leading to stimulation of glucose transport and activation of the mitogen-activated protein kinase cascade. The active insulin receptor phosphorylates tyrosine residues of intracellular proteins such as the insulin receptor substrate-1 (IRS-1) which acts as docking sites for molecules containing Src homology 2 (SH2) domains. One such molecule is phosphatidylinositol 3-kinase (PI 3-kinase) which becomes activated by binding to IRS-1. PI 3-kinase activity is required for the insulin-stimulation of glucose transport and glycogen synthesis. Grb2, a small adaptor molecule, can bind IRS-1 and, through the guanine nucleotide exchange factor Sos, leads to the activation of the small GTP binding protein Ras. Through a cascade of protein kinases, activation of Ras results in activation of the Erk 1 and 2 mitogen-activated protein kinases (MAPKs) which appear to control important nuclear and metabolic events. To investigate the role of the actin network in the propagation of insulin action leading to stimulation of glucose transport and the activation of the Erk MAPKs, we used the fungal metabolite cytochalasin D which disassembles the actin network. Actin disassembly abolished almost completely the ability of insulin to increase the rate of glucose transport into L6 muscle cells (myotubes) through prevention of the insulin-induced recruitment of glucose transporters to the plasma membrane which is the event that mediates the increase in the rate of transport. Actin disassembly did not affect either the insulin-mediated phosphorylation of IRS-1, the association of PI 3-kinase with this molecule, or the activation of IRS-1-associated PI 3-kinase. These results were also verified in another insulin responsive cell line, the 3T3-L1 adipocytes. In these cells, actin disassembly inhibited the insulin-induced recruitment of PI 3-kinase to intracellular membranes containing glucose transporters. Moreover, actin disassembly abolished the insulin-mediated phosphorylation of the Erk MAPKs. We conclude that the cellular actin network of insulin responsive cells is not required for the activation of PI 3-kinase but prevents its cellular redistribution. In contrast, intact actin filaments are essential for the propagation of insulin signals leading to the the activation of the MAPKs. PMID- 9210238 TI - The mandibular marginal ramus of the facial nerve: an anatomic and clinical study. AB - Injury to the mandibular marginal ramus of the facial n. constitutes a risk in cervicofacial surgery. The aims of this study were to define the origin of this nerve branch and its course and relations, especially with the lower border of the mandible and the facial vessels. Our observations revealed differences from the classical description of a single nerve branch traveling on the outer aspect of the body of the mandible above its lower border. We found several marginal branches, which become closely related to the facial pedicle, particularly the intermediate ramus, which can form a neural plexus around the facial a. They may follow a submandibular course, before but also after crossing the facial vessels. They are difficult to classify because of their great variability. PMID- 9210239 TI - Facial vein terminating in the external jugular vein. An embryologic interpretation. AB - The external jugular v. (EJV) is increasingly being used for therapeutic procedures and monitoring by clinicians. In view of this clinical relevance, dissection was done on the head and neck regions in 40 adult cadavers of Indian origin to detect variations of the EJV. Though several patterns of tributaries were found, a facial v. (FV) of considerable size was observed coursing obliquely to join the EJV in the neck in four cases (5%). The distance of the junction of the FV and the EJV from the angle of the mandible ranged between 55 and 104 mm. This may represent a persistent communication of the primitive linguofacial v. with the secondarily developing EJV. This anastomotic channel is present for some time in the fetus but later undergoes retrogression. Its persistence in some individuals results in this variation. PMID- 9210240 TI - Venous drainage of the dorsal sector of the liver: differences between segments I and IX. A study on corrosion casts of the human liver. AB - The aim of this study was to determine the venous drainage of the dorsal sector of the liver in order to define the differences between segments I and IX and their implications for sectorially and segmentally oriented hepatic surgery. The study was based on corrosion casts of 61 macroscopically healthy livers. The drainage pathways of veins at least 10 mm long and 1 mm wide were evaluated and statistically analysed. On average, 9 veins drained the two segments and three veins from both segments entered the inferior vena cava. In 95% of cases the veins from segment I drained predominantly into the inferior vena cava, whereas in segment IX this pathway was dominant in only 30% of cases. In 64% of cases a vein originating in segment IX entered the right hepatic v. The difference in the venous drainage of the two segments suggests that segment IX partly belongs to the neighbouring segments and may thus be only a paracaval region of the right liver. PMID- 9210241 TI - Acetabular anatomy and the relationship with pelvic vascular structures. Implications in hip surgery. AB - Most direct vascular trauma occurring during hip surgery results from injury to pelvic vascular structures which are not visible during the procedures of reaming, drilling holes or the fixation of screws. In this study, 5 pelves of fresh cadavers were injected with a radiopaque mixture and were visualised with a scanner according to 5 predetermined sections. Bone depth of the acetabulum was measured in each section. A calculation was made describing the minimal distance separating the inner cortex from the principal pelvic vessels. After an anatomic dissection of each pelvis, the relationship between the vessels and screws of the fixation cup, implanted identically on the quadranted acetabulum, was observed. The screws placed in the anterior and inferior quadrants and the center of the acetabulum endangered the external iliac v. and a. and the obturator pedicle. The depth of the periacetabular bone was greater in the superior and posterior quadrants. The inferior gluteal, pudendal and superior gluteal aa. were more than ten mm from the posterior wall. Conversely, the external iliac and obturator pedicles came in contact with the osseous surface on which they lay. A projection of the vessels on the acetabulum was made, and the reproducible character of the acetabular-quadrant system was verified. The superior quadrant offers all the characteristics of a vascular safe zone. A knowledge of these anatomic relationships explain vascular trauma in pelvic fractures and helps to prevent vascular injury in hip surgery. PMID- 9210242 TI - Evolution of the angle of obliquity of the femoral diaphysis during growth- correlations. AB - Numerous studies of the bicondylar angle of the adult femur have been carried out in human anatomy, paleoanthropology and primatology. The aim of this paper is to study the evolution of this angle in relation to age and acquisition of walking in young children. Seventy-seven radiographs of children, ranging from 5 months to 17 years postnatally, and of four dead newborn were analysed. The measurements concern the bicondylar angle (A.O.F.), the collo-diaphyseal angle (A.C.D.), the length of the femoral neck (L.N.) and of the femur (L.F.) and the interacetabular distance (D.I.A.). Some children were x-rayed at different ages, which permits a longitudinal as well cross-sectional study. The results show that there is no sexual dimorphism and that the increase in the angle is closely related to the age of the child. The bicondylar angle starts at 0 degree at birth and then increases progressively with growth to reach adult values of at least 6 degrees-8 degrees between 4 and 8 years postnatally. In adults, the mean values are between 8 degrees and 11 degrees and the maximum range is between 6 degrees and 14 degrees. The obliquity angle corresponds to an angular remodeling of the femoral diaphysis, which is independent of the growth and shape of the distal femoral epiphysis. The tibio-femoral angle measures the evolution of a physiologic phenomenon, from the load "in varus" to the load "in valgus" of the lower limb. It is linked with the bicondylar angle but is different from it. PMID- 9210243 TI - Anatomical variants of the profunda femoris artery: an angiographic study. AB - The bifurcation of the common femoral artery (CFA) into superficial and profunda femoris arteries (PFA) and the branching pattern of the PFA are subject to considerable normal anatomical variation. These variation patterns were assessed on normal lower limb angiograms of 94 adult patients. The main pattern (in 81% of patients) consisted of both medial and lateral circumflex arteries arising directly from a PFA situated lateral or posterolateral to the femoral artery. Previous studies of arterial variations in this region and the relevant embryology are reviewed. Relevance to angiographic procedures of the lower limb is discussed. PMID- 9210244 TI - The fornix of the human brain: evidence of left/right asymmetry on axial MRI scans. AB - This article reports the observation that there is a left/right asymmetry of the anterior columns of the fornix in the human brain. This asymmetry is present in the position of the two columns of the fornix in relation to the septum pellucidum. The left columna fornicis was found to be located caudal to the right, and this can be readily visualized on axial MRI scans. This difference was seen in most of the subjects, but in some subjects there was no left/right difference and in a few the asymmetry was inverse. The asymmetry of the fornix with respect to the anterior-posterior axis was independent of the well-known dissimilar lateral ventricular volumes. However, the left/right difference in the position of the fornix was evident in subjects with or without differences in ventricular volumes. This suggests that the mechanism underlying the development of asymmetry of the fornix is independent of the mechanism leading to ventricular asymmetry. So far, no functional relevance has been ascribed to such differences in location. The finding is gaining interest in connection with recent reports of asymmetries in hippocampal subfields. Studies of fornical lesions should therefore give attention to possible side-to-side differences. PMID- 9210245 TI - Anatomic and MRI study of the subtalar ligamentous support. AB - The diagnosis of subtalar instability remains difficult both clinically and radiographically. The authors present an anatomic and MRI study of the subtalar ligamentous support. The anatomic study has consisted in dissections and sections of cryoconserved hindfeet (15 cases) which precises the organisation of ligamentous bundles in the lateral (sinus tarsi) and central (canalis tarsi) subtalar compartments, mainly represented by the trilayered inferior extensor retinaculum, the cervical talo-calcaneal ligament and the interosseous talo calcaneal ligament. MRI study (1.5 tesla) of anatomic specimens was performed according to defined types of sections: sagittal, coronal, coronal oblique, axial transverse. The correlations of anatomic and MRI sections allowed a precise interpretation of the subtalar ligamentous support as anatomically described. A complementary clinical MRI study was performed which allowed the validation of "the inversion test": this test optimizes the visualization of the different ligamentous structures. Relative to the difficulties of conventional imaging procedures, MRI appears of clinical relevance in the diagnosis of subtalar instabilities. This technique allows direct visualization of ligaments (or their rupture) and therefore a better evaluation of subtalar involvement in ankle sprain. This paper present a functional concept in MRI articular ligamentous restraints concern. PMID- 9210246 TI - An in vitro anatomic model of the human cerebral arteries with saccular arterial aneurysms. AB - An in vitro model of the main human cerebral arteries with or without saccular arterial aneurysms is presented. A cast of the cerebral arteries was obtained in a human specimen. Three aneurysms were simulated and added to the cast. Wax copies of the cast were produced, and embedded with liquid resin solidifying into solid blocks. After evacuation of the wax, a model consisting of a hollow reproduction of the cast within the resin block was obtained. The model is reproducible and anatomically accurate. Since it is transparent to visible light, and compatible with x-ray, magnetic resonance and transcranial doppler techniques, it should prove useful for a wide range of haemodynamic and radiologic investigations. The reported technique may be adapted to any structure with a hollow configuration, allowing for the preparation of arterial and venous models from other vascular areas, as well as models from other anatomic systems, such as the biliary or urinary tracts. PMID- 9210247 TI - Complex variation of the superficial palmar arch--case report. AB - This article describes a complex variation in the pattern of blood supply to the palm of the hand. In the present case; a) the superficial palmar branch of the radial a. coursed superficial to the thenar mm.; b) The princeps pollicis and radialis indicis aa. arose from the superficial palmar branch of the radial a.; c) The first and second common palmar digital aa. arose as a common trunk from the superficial palmar arch. PMID- 9210248 TI - Clinical and molecular analysis of neurodegenerative diseases. AB - Clinical and molecular analyses of neurodegenerative diseases such as Alzheimer's disease (AD), amyotrophic lateral sclerosis (ALS), and spinocerebellar ataxia type 1 (SCA1) were performed. In the present study, a Japanese family of AD with an Ala285Val substitution in exon 8 of the presenilin-1 (PS-1) gene was found. This family was characterized by relatively late onset (mean age at 50 years) in familial AD with PS-1 gene mutation and by absence of myoclonus, seizure or paratonia. Magnetic resonance image (MRI) study showed marked linear signal abnormalities in white matter of parietoocctipital lobes, suggesting a presence of cortical amyloid angiopathy of the patient with PS-1 gene mutation. Clinical characteristics of familial amyotrophic lateral sclerosis (FALS) with four different missense point mutations in exons 2, 4, and 5 of the Cu/Zn superoxide dismutase (SOD) gene were reported. Although features of progressive neurogenic muscular atrophy was common in patients of these families, patients of each family showed characteristic clinical features. Although lower motor sign was evident in all cases, hyperreflexia varied from 0 to 100% among patients with the different mutations, and Babinski sign was not observed in any cases. Bulbar palsy was frequent with a mutation, but not present with another mutation. SOD activity of red blood cells was generally reduced with minor variations. CAG trinucleotide repeat expansion was analyzed in 25 families with hereditary ataxia of Menzel type in the northeast of Japan. Twenty of 38 patients in 12 families had expanded allele for spinocerebellar ataxia type 1 (SCA1). Study of the number of CAG repeats in various tissues showed no differences in the repeat length in lymphocytes, muscle or brain; sperm, however, showed an obvious expansion. This may be a clue to a possible mechanism for the molecular basis of paternal anticipation of the disease. These results suggest that clinical features of some familial cases of neurodegenerative diseases such as AD, ALS, and SCA1 are well correlated with their genetic mutations. PMID- 9210249 TI - Kidney and hypertension: role of the juxtaglomerular apparatus. AB - The kidney plays an important role in the pathophysiology of hypertension. Recent studies suggest that glomerular hemodynamics may be critically involved not only in the pathogenesis of hypertension but also in the mode of progression of renal dysfunction. The juxtaglomerular apparatus (JGA), consisting of the glomerular afferent and efferent arterioles and the specialized tubular epithelial cells called the macula densa, plays a central role in the regulation of glomerular hemodynamics and renin release. This article reviews the mechanism by which the JGA controls renin release and glomerular hemodynamics as well as its relevance in the pathogenesis, pathophysiology and treatment of hypertension. PMID- 9210250 TI - Effect of interferon-gamma on lymphocyte cell subsets in human large bowel: a study using organ culture method. AB - This study was conducted to investigate the effects of interferon (IFN)-gamma on normal colonic lamina propria lymphocyte subsets in humans using organ culture method. Lamina propria lymphocyte subsets in normal colonic biopsy tissues receiving 1 x 10(5) u/ml of IFN-gamma (IFN-gamma-treated group) were investigated in comparison with those cultured in medium only (IFN-gamma-non-treated group) for 24 hr. CD8-positive cells and IgG, IgA1 and IgM-containing cells were elevated in the IFN-gamma-treated group compared with those in the IFN-gamma-non treated group, which was similar to immunological changes in mucosal lesions of inflammatory bowel disease. PMID- 9210251 TI - 4 Pro-R hydrogen of NADPH was abstracted for enzymatic hydride transfer by N ethylmaleimide-reductase of Yarrowia lipolytica. AB - We studied the steric course of the reaction catalyzed by the N-ethylmaleimide (NEM) reductase of Yarrowia (Candida) lipolytica (Y. lipolytica), using 4R-[4 2H1]NADPH and 4S-[4-2H1]NADPH as cofactors and N-ethylcitraconimide as substrate. Active substrates and inhibitors of NEM reductase and its subcellular distribution were also investigated to clarify the biochemical properties of this enzyme. NEM reductase catalyzes the reduction of N-ethylmaleimide to N ethylsuccinimide with NAD(P)H as the cofactor. Several maleimide and cyclopentenone derivatives tested were also active substrates for NEM reductase of Y. lipolytica. Some pyrazolone derivatives, particularly 1-phenyl-5 pyrazolone, were found to be effective inhibitors of NEM reductase. Subcellular localization of NEM reductase was carried out using protoplast formation and differential centrifugation. Ninety-eight percent of the NEM reductase activity was recovered in the cytosolic fraction, indicating that NEM reductase in Y. lipolytica was the cytosolic enzyme. We also determined the stereochemical specificity of the reduction of N-ethylcitraconimide by NEM reductase in Y. lipolytica, showing that 4 Pro-R hydrogen of NADPH was abstracted for enzymatic hydride transfer by NEM reductase, and two hydrogen atoms from NADPH and H2O added to opposite faces of the double bond of N-ethylcitraconimide. PMID- 9210252 TI - Management of functional pulmonary atresia with isoproterenol in a neonate with Ebstein's anomaly. AB - Ebstein's anomaly is a rare congenital cardiac anomaly showing significant clinical manifestations with a high mortality rate in the neonatal period. The prognosis of the patient is essentially determined by the severity in morphological changes, however, high pulmonary vascular resistance in the neonatal period may aggravate tricuspid regurgitation and lead to functional pulmonary atresia. We describe a critically ill neonate with morphologically mild Ebstein's anomaly who was successfully managed with intensive care including isoproterenol administration for functional pulmonary atresia. Isoproterenol is a potent pulmonary vasodilator with inotropic and chronotropic effects, and seemed to decrease the pulmonary vascular resistance allowing increased antegrade blood flow to the pulmonary artery and improved cardiac output. If tachycardia is not present, isoproterenol administration is recommended in critically ill neonates with anatomically mild Ebstein's anomaly and no associated cardiac defects. PMID- 9210253 TI - An evaluation of neuromuscular reversal with edrophonium in a patient with malathion intoxication. AB - We evaluated the neuromuscular reversal with edrophonium using peripheral nerve stimulator and recorder in a patient with malathion intoxication. Edrophonium 10 mg i.v. caused an increase in single twitch tension by 76% of the control during the recovery phase from an acute cholinergic crisis 16 days after ingestion of malathion solution. The present study indicated that edrophonium test seems to be a reliable monitoring in evaluating neuromuscular reversal in the patient with acute malathion insecticide poisoning. PMID- 9210254 TI - Neuromuscular blocking actions of the aminoglycoside antibiotics sisomicin and micronomicin in the rabbit. AB - The neuromuscular blocking actions of sisomicin sulfate (SISO), micronomicin sulfate (MCR) and d-tubocurarine (dTc) were studied in 20 rabbits anesthetized with halothane. The i.v. administration of SISO 20-40 mg/kg, MCR 40-80 mg/kg or dTc 0.1-0.3 mg/kg resulted in dose-dependent decreases in twitch tension. The ED50s for SISO, MCR and dTc were 23.5, 58.2 and 0.2 mg/kg, respectively. SISO- and MCR-induced neuromuscular blockade was partially antagonized by neostigmine or by calcium. PMID- 9210255 TI - Gadolinium chloride toxicity in the rat. AB - Groups of 10 male and 10 female Sprague-Dawley rats were given a single intravenous injection of gadolinium chloride solution at dosages of 0 (saline vehicle), 0.07, 0.14, and 0.35 mmol/kg. Apart from 1 top-dose female, which died during dosing, 5 rats/sex/ group were necropsied 48 hr postdose, and the remaining 5 rats/sex/group were necropsied 14 days postdose. Macroscopic, hematological, and clinical chemistry analyses were undertaken on all animals that were necropsied. Histopathological examination was undertaken on all organs from high-dose and control animals necropsied 48 hr postdose and on tissues that showed treatment-related changes from all other rats necropsied either 48 hr or 14 days postdose. Major lesions related to gadolinium chloride administration consisted of mineral deposition in capillary beds (particularly lung and kidney), phagocytosis of mineral by the mononuclear phagocytic system, hepatocellular and splenic necrosis followed by dystrophic mineralization, mineralization of the fundic glandular mucosa in the absence of necrosis followed by mucous cell hyperplasia, decreased platelet numbers and increased prothrombin time, and activated partial thromboplastin time. Electron microscopy and x-ray microanalysis of the spleen and liver revealed electron-dense deposits in splenic macrophages, Kupffer cells, and hepatocytes composed of gadolinium, calcium, and phosphate. PMID- 9210257 TI - The effect of erythritol on the stability of gamma-glutamyl transpeptidase and N acetyl glucosaminidase in human urine. AB - In toxicology studies and clinical trials of erythritol, treated animals and human subjects had higher urine gamma-glutamyl transpeptidase [gamma-glutamyl transferase (gamma-GT)] than untreated controls. It has previously been reported that gamma-GT activity in frozen urine decreases with time; therefore, a study was undertaken to examine the effects of storage temperature, time, and the presence of erythritol on the stability of gamma-GT and N-acetyl glucosaminidase in human urine. In this study, it was found that the rate of decrease of the activity of gamma-GT is much greater at -20 degrees C than at -70 degrees C. Variation in the storage temperature of the frozen urine is particularly deleterious to gamma-GT. The addition of erythritol in a concentration of 5% reduces this decrease. Approximately 15% of N-acetyl glucosaminidase activity is lost in the initial process of freezing the urine. Thereafter, conditions of temperature, time, and the presence or absence of erythritol account for little additional loss of activity. PMID- 9210256 TI - Body weight-tumor incidence correlations in long-term rodent carcinogenicity studies. AB - Associations between body weight and tumor incidence and among the incidences of selected site-specific tumors were examined for more than 4,000 male and female Fischer-344 rats and B6C3F1 mice in the National Toxicology Program historical control database. Incidences of certain site-specific tumors, most notably mammary gland and pituitary gland tumors in rats and liver tumors in mice, were shown to have a strong positive correlation with 52-wk body weight. Using individual animal data, logistic regression models were derived for predicting site-specific tumor incidence as a function of 52-wk body weight, age, and other factors. This association between body weight and tumor incidence can explain many of the decreased tumor incidences observed in National Toxicology Program carcinogenicity studies. Body weight differences between dosed and control groups can also mask carcinogenic effects for those sites sensitive to body weight changes. Thus, when designing long-term rodent carcinogenicity studies, measures should be taken to minimize potential body weight differences between dosed and control groups. There were a number of small but significant negative correlations among tumor incidences, reflecting primarily the lethality of the tumors in question. None of these correlations (nor the 2 small positive correlations found) are likely to have any impact on the interpretation of experimental results. However, the high negative correlation between pituitary gland and testis tumors in male Fischer-344 rats cannot be dismissed so easily, does not reflect tumor lethality, and is currently being studied further. PMID- 9210258 TI - Mesenchymal tumors of the mouse urinary bladder with vascular and smooth muscle differentiation. AB - Bifenthrin, a synthetic pyrethroid insecticide/miticide, has been fed to male and female Swiss Webster mice at levels of 0, 50, 200, 500, and 600 ppm in the diet for between 604 and 644 days. Tumors of the urinary bladder were observed and initially reported as leiomyosarcomas. Subsequently, the bladders were reviewed and the tumors showed a pattern of both epithelioid cells and spindle cells forming irregular vascular channels. The tumors appeared to arise from the trigone of the bladder and, in some cases, invaded the bladder wall. No metastases were recorded. The tumor is usually considered rare; however, in this study, it was commonly observed in all groups but predominantly in males. The histogenesis of the tumor is uncertain, but from its pleomorphic histological features, including smooth muscle and vascularity, it is probably derived from vascular mesenchyme. PMID- 9210260 TI - Dose-dependent amplification by L-ascorbic acid of NaHCO3 promotion of rat urinary bladder carcinogenesis. AB - The dose dependence of L-ascorbic acid (AsA) copromotion of urinary bladder carcinogenesis with continuous concomitant administration of NaHCO3 was investigated. In the first experiment, 83 male F344 rats were all given 0.05% N butyl-N-(4-hydroxybutyl)nitrosamine (BBN) for 4 wk and then divided into 5 groups, which received basal diet (Oriental MF) containing AsA at 0, 1, 2, 3.5, or 5% plus 1.5% NaHCO3 for 32 wk. Relative urinary bladder weights in the 5% AsA group were significantly increased as compared to the 0 or 1% group values due to the development of tumors. Both the incidence and number of microscopic urinary bladder lesions (tumors and preneoplastic lesions) showed dose-dependent increases. Furthermore, the sizes of the urinary bladder tumors (carcinomas and papillomas) were significantly increased with the highest dose, 5-bromo-2' deoxyuridine labeling indices showed slightly increased proliferation in preneoplastic lesions of the urinary bladder epithelium with 5% AsA treatment. In a separate experiment, scanning electron microscopic observation revealed that administration of 5% AsA plus 1.5% NaHCO3 for 8 wk, without BBN, altered the urinary bladder surface. Elevation of urinary bladder epithelium AsA content, as well as urinary AsA, was also noted. Ornithine decarboxylase (ODC) activity and ODC messenger RNA levels in urinary bladder epithelium of rats treated with 1.5% NaHCO3 plus 5% AsA for 8 wk showed no statistically significant differences as compared to the control group. The results indicate that AsA amplifies the rat urinary bladder carcinogenesis promotion activity of NaHCO3 and that its intensity of action depends on the dose, particularly at high dose. PMID- 9210259 TI - Quantitative relationship between transforming growth factor-alpha and hepatic focal phenotype and progression in female mouse liver. AB - Modulations in the positive hepatocyte growth factor, transforming growth factor alpha (TGF-alpha) and its receptor epidermal growth factor receptor (EGFR), occur in rat and human liver tumors. The purpose of this study was to determine if TGF alpha and EGFR are altered in basophilic and acidophilic preneoplastic and neoplastic liver lesions generated in DEN-initiated mice exposed to a variety of hepatocarcinogens. Female B6C3F1 mice were initiated with N-nitrosodiethylamine (DEN) and treated with hepatocarcinogenic concentrations of unleaded gasoline vapor (2,000 ppm), methyl tertiary butyl ether vapor (7,814 ppm), phenobarbital (500 ppm, diet), or chlordane (25 ppm, diet). Hepatic foci and tumors were identified and evaluated immunohistochemically with antibodies for TGF-alpha and EGFR. In all treatment groups, basophilic hepatic foci were negative for TGF alpha immunoreactivity (554/564, 98%). In contrast, regardless of treatment, acidophilic hepatic foci were immunoreactive for TGF-alpha (107/108, 99%). There was no significant difference in mean hepatic labeling index as measured by the incorporation of 5-bromo-2'-deoxyuridine between foci immunoreactive and nonimmunoreactive for TGF-alpha. The incidence of immunoreactivity for TGF-alpha increased in hepatocellular tumors that were predominantly of the basophilic phenotype. Of basophilic hepatocellular adenomas, 16/81 (20%) were immunoreactive for TGF-alpha, while 17/29 (59%) of hepatocellular carcinomas stained positive for TGF-alpha. A similar increased incidence of EGFR immunoreactivity was found in basophilic hepatocellular adenomas (17/67, 25%) and carcinomas (19/28, 68%) relative to basophilic foci (11/367, 3%), suggesting an autocrine mechanism for the development of mouse liver tumors. The increased incidence of TGF-alpha immunoreactivity in basophilic liver tumors suggests that TGF-alpha is a marker of tumor progression in mouse liver. Furthermore, TGF-alpha modulations were dependent on phenotype rather than treatment, indicating inherent differences in the expression of TGF-alpha in basophilic and acidophilic hepatic lesions. PMID- 9210261 TI - A brief review of formaldehyde carcinogenesis in relation to rat nasal pathology and human health risk assessment. AB - The 1980 report that inhaled formaldehyde induced nasal squamous cell carcinomas in rats had a significant societal impact and resulted in extensive research in the fields of rodent nasal pathology and human cancer risk assessment. This article presents an overview of the evolution of these events. It is concluded that the nasal passages of humans and rats are fundamentally identical biological target organs. Nevertheless, in the case of human health risk assessment, minor differences between these species may be critically important. Special attention should be paid to interspecies differences in nasal dosimetry and local metabolism; thus, chemical toxicity data derived from rats require careful interpretation when used for human risk assessments. In the case of formaldehyde, it is recommended that low-concentration (< or = 2 ppm airborne exposure) extrapolation, where no tissue damage is observed, be uncoupled from the responses at high concentrations (> or = 6 ppm), where epithelial degeneration, regenerative cell replication, and inflammation appear to be essential driving forces in formaldehyde carcinogenesis. The presence of treatment-related nasal lesions in rats following exposure to chemicals should always be treated as an indication of a potential human health risk, whether exposure is by the inhalation, oral, or dermal route. PMID- 9210262 TI - Spontaneous iron overload in Sprague-Dawley rats. AB - In a review of the toxicological studies performed in our laboratory during the period 1986-1995, we occasionally observed significant iron overloading in the liver. Liver tissue was examined by light and electron microscopy, and the results were analyzed by sex and age (7, 9, 11, 19, 31, 59, and 111 wk). The intensity of iron overload increased with age: the accumulation began in pericanalicular siderosomes of periportal hepatocytes and extended progressively to the entire lobule and also to nonhepatocytic cells (Kupffer cells in sinusoids and macrophages around bile ducts in portal tracts) and occasionally with distortion of sinusoids by sideroblastic nodules and moderate enlargement of portal tracts in the oldest animals. No significant inflammatory infiltrates, degeneration, necrosis or fibrosis were noted. Hepatocyte pigmentation alone was prominent at 9 and 11 wk. The frequency of pigmentation of parenchymal and nonhepatocytic cells increased from 9 wk for females; in males, this was seen only at 111 wk. The intensity of pigmentation of nonhepatocytic cells versus parenchymal cells increased with aging. The frequency of those different types of iron overloading was higher for females up to 111 wk. The pathology of spontaneous iron overloading in the Sprague-Dawley rat, described here in spite of differences, has some similarities to that of human hereditary hemochromatosis. PMID- 9210263 TI - A spontaneous testicular teratoma in an ICR mouse. AB - A spontaneous teratoma was detected in the left testis of a 7-wk-old male ICR (Crj:CD-1) mouse. The tumor was composed of a variety of well-differentiated tissue elements from all 3 germ layers including neuronal tissue and stratified epithelium as ectoderm-derived tissues, bone, cartilage, striated muscle, and adipose tissue as mesodermal tissues, and respiratory columnar epithelium as endoderm-originated tissue. No tumor metastasis was detected. It may be of congenital origin as in strain 129 mice. To the best of our knowledge, this is the first report concerning spontaneous testicular teratoma in a strain ICR mouse. PMID- 9210264 TI - Appearance of eosinophilic granule cells as pathological changes in the uteri of two aged virgin Donryu rats. AB - The morphological features of cells containing eosinophilic granules (EG cells) present as pathological changes in the uteri of 2 aged virgin Donryu rats were described and compared with granulated metrial gland (GMG) cells of a pregnant rat. Microscopically, EG cells distributed diffusely from the mesometrial triangle to the subendometrium with partly focal accumulation in Case 1 and infiltrated diffusely from the endometrium to the serosa without any clumping in Case 2. They were large cells with eosinophilic and hyaline intracytoplasmic granules, positive for the periodic acid-Schiff reaction. Both EG and GMG cells reacted positively to concanavalin A, wheat germ agglutinin, and phytohemagglutinin on lectin histochemical investigation, although common results were not always obtained for immunohistochemistry. These results show that EG cells are morphologically similar to GMG cells, suggesting a possibility that EG cells are GMG cells. EG cells were without neoplastic properties or pronounced cell-proliferative activity. In Case 1, histological changes of the ovaries and vagina were indicative of a pseudopregnant status. In Case 2, the rat had been maintained under abnormal hormonal conditions, and no evidence of any pseudopregnant-like status was found. These results suggest, thus, that the appearance of EG cells is evidence of a reactive pathological condition in the uteri of aged virgin rats, although the precise pathogenesis and biological significance of EG cells remain to be determined. PMID- 9210265 TI - Mixed mesenchymal tumor in the kidney of a young beagle dog. AB - A rare spontaneous tumor was detected in the kidney of an 11-mo-old beagle dog. The tumor was a well-circumscribed cortical mass and was composed of a heterogeneous population of connective tissue cell types. While the primary cell type was a spindle cell associated with prominent collagen deposition, other areas contained bands of smooth muscle cells and poorly cellular myxomatous tissue. The presence of smooth muscle cells was confirmed by transmission electron microscopy and immunoperoxidase techniques. Based on the results, the tumor was diagnosed as a benign mixed mesenchymal tumor. PMID- 9210266 TI - The Min mouse: a genetic model for intestinal carcinogenesis. PMID- 9210267 TI - "Have you seen this?" Pituitary cysts in a mouse. PMID- 9210268 TI - Cell transformation assays as predictors of carcinogenic potential. PMID- 9210269 TI - Pathology peer review. PMID- 9210270 TI - Pathology peer review. PMID- 9210271 TI - Pathology peer review. PMID- 9210272 TI - Men who batter: recent history and research. PMID- 9210273 TI - A large sample empirical typology of male spouse abusers and its relationship to dimensions of abuse. AB - A number of studies have described typologies of domestically violent men. Holtzworth-Munroe and Stuart (1994) recently proposed a theoretical model for predicting violence severity and generality from personality "type." The present study, using data from 833 identified abusive men, tested the model. Personality types were determined from cluster analysis of data from the Millon Clinical Multiaxial Inventory, and resulted in a three-cluster solution consistent with the Holtzworth-Munroe and Stuart model. The three main clusters included nonpathological, antisocial, and passive aggressive-dependent groups. Three other, smaller types were also identified. Multivariate and chi-square analyses comparing the main clusters on other variables generally supported the Holtzworth Munroe and Stuart model. Nonpathological men had the lowest maximum violence and frequency. They restricted their violence primarily to intimate relationships and had the fewest police contacts. Antisocial and passive aggressive-dependent men did not differ in maximum violence, but antisocial men were the most generally violent and had the most police contacts. Passive aggressive-dependent men had the highest frequency of violence. Clinical, theoretical and methodological implications are discussed. PMID- 9210274 TI - Generalized versus spouse-specific anger/hostility and men's violence against intimates. AB - The present study examined the extent to which generalized versus spouse-specific anger/hostility was associated with partner violence in 263 men seeking conjoint marital therapy. Clinic men were classified as nonviolent (NV), moderately violent (MV), and severely violent (SV). A community comparison group of relationship-satisfied, non-violent men (CO) was also included. All clinic men reported higher levels of generalized and spouse-specific anger, spouse-specific aggression/hostility, depressive symptomatology and lower spouse-specific assertiveness than community men. SV men reported higher levels of spouse specific anger/hostility, relationship discord, depressive symptomatology, and lower general problem-solving ability than NV men. Regression analyses confirmed that spouse-specific anger/hostility, low problem-solving ability, and relationship discord were significant predictors of men's violence. Overall, generalized anger and hostility were not unique predictors of men's violence against intimates. PMID- 9210275 TI - Comparing the emotional reactions and behavioral intentions of violent and nonviolent husbands to aggressive, distressed, and other wife behaviors. AB - The present study was designed to compare the self-reported emotional reactions and behavioral intentions of violent and nonviolent husbands to a variety of wife behaviors depicted with standardized stimuli. We recruited four subject groups, including 25 men beginning domestic violence treatment programs and three groups from the community-21 maritally violent and maritally distressed men, 23 nonviolent/distressed men, and 28 nonviolent/nondistressed men. Using stimuli and measures derived from Biglan, Rothlind, Hops, and Sherman (1989), along with stimuli designed for this study, subjects read written descriptions and examples of various wife statements (e.g., aggressive, distressed, and facilitative) and viewed videotaped depictions of wife behaviors varying in verbal content and nonverbal affect (i.e., aggressive/irritated, aggressive/dysphoric, distressed/irritated, and distressed/dysphoric). In response to each wife behavior, subjects rated what their emotional reactions (e.g., sympathetic, caring, supportive, sad, anxious, irritated, angry) and their behavioral responses (e.g., try to comfort, say something supportive, discuss the subject, not say anything, say something hostile, argue) would be. As predicted, in response to a wide range of wife behaviors, and relative to nonviolent men, violent men were less likely to report sympathetic/positive emotions and more likely to experience anger and irritation, but not other negative (i.e., sad or anxious) emotions. They were also more likely to report negative behavioral intentions and less likely to report positive behavioral intentions. The theoretical and clinical implications of these findings are discussed. PMID- 9210276 TI - The applicability of the theory of planned behavior to abusive men's cessation of violent behavior. AB - This study examined the ability of Ajzen's (1988; 1991) Theory of Planned Behavior (TPB) to explain men's cessation of violent behavior. TPB suggests that a man's intention to abuse his female partner, and therefore his subsequent abusive behavior will be determined by: (1) his evaluation of possible outcomes of abusive behavior (attitudes toward behavior); (2) his perception of the expectations of others around him concerning violence and (3) the degree to which he believes he can control his abusive behavior. Pretest self-report measures from men and follow-up recidivism data based on partner report were available for 176 cases drawn from a previous study conducted by Harrell (1991). Reliable proxy measures for TPB variables (intentions/expectations to use violence, attitudes toward behavior, social norms, perceived behavioral control) were created. Regression analyses testing the TPB model provided modest support for prediction of intention to reabuse and subsequent abusive behavior. Of the TPB variables, perceived control appeared to be most important in understanding batterers' intentions to abuse and their subsequent abusive behavior. Refinement in measurements and the need for additional modifications to the model are discussed. PMID- 9210277 TI - Desistance of husband aggression in the early years of marriage. AB - Desistance of husband marital aggression was investigated over a 3-year time span. One hundred and eighty-eight couples who had experienced husband aggression in the first year of marriage were followed to the third year of marriage. Overall 23.9% of these husbands had no violence in years 2 and 3. However, desistance rates differed as a function of the type of violence that occurred in year 1. Husbands who had only one incident of minor physical aggression and no severe violence in year 1 were most likely to desist in years 2 and 3 while those who used severe violence in year 1 were least likely to desist. Subsequent analyses showed that wife's depression and dissatisfaction with the partner increased from years 1 to 3 when desistance did not occur. PMID- 9210278 TI - Psychological factors in the longitudinal course of battering: when do the couples split up? When does the abuse decrease? AB - The longitudinal course of battering was investigated over a 2-year time span. Forty-five batterers and their spouses were assessed with self-report, psychophysiological, and marital interaction measures. Both the stability of the relationship and of the battering were assessed. At the two-year follow-up, 62% of the couples were still married and living together, while 38% had separated or divorced. A combination of six variables, reflecting severity of husband emotional abuse, wife dissatisfaction, husband physiological arousal, and wife defending herself assertively, was 90.2% accurate in predicting separation or divorce 2 years later. Of the couples still living together at follow-up, 46% of the batterers did not reduce their levels of severe violence, while 54% did significantly decrease levels of violence. Husbands who continued to be severely violent at 2-year follow-up were more domineering, globally negative and emotionally abusive toward their wives at Time 1 than husbands who reduced their levels of violence. Even though 54% of the batterers decreased the frequency of violent acts over the 2-year period, only 7% achieved complete desistance. Moreover, husband emotional abuse did not decrease over the 2-year period, even when physical abuse did. PMID- 9210279 TI - Feminist-cognitive-behavioral and process-psychodynamic treatments for men who batter: interaction of abuser traits and treatment models. AB - At a community-based domestic violence program, 218 men with a history of partner abuse were randomly assigned to either feminist-cognitive-behavioral or process psychodynamic group treatments. The treatments were not hypothesized to differ in outcome. However, men with particular characteristics were expected to have lower recidivism rates depending on the type of treatment received. Treatment integrity was verified through audio-taped codings of each session. The partners of 79% of the 136 treatment completers gave reports of the men's behavior an average of 2 years post-treatment. These reports were supplemented with arrest records and self-reports. Rates of violence did not differ significantly between the two types of treatment nor did reports from the women of their fear level, general changes perceived in the men, and conflict resolution methods. However, interaction effects were found between some offender traits and the two treatments. As predicted, men with dependent personalities had better outcomes in the process-psychodynamic groups and those with antisocial traits had better outcomes in the cognitive-behavioral groups. The results suggest that more effective treatment may occur if it is tailored to specific characteristics of offenders. PMID- 9210280 TI - Impaired mitogen-driven proliferation and cytokine transcription of lymphocytes from macaques early after simian immunodeficiency virus (SIV) infection. AB - Altered cytokine transcription might play an important role in the pathogenesis of human immunodeficiency virus (HIV) infection in humans. The infection of rhesus macaques with simian immunodeficiency virus (SIV) provides a relevant animal model for HIV infection. Therefore, we evaluated the cyokine transcription of phytohemagglutinin (PHA)-stimulated lymphocytes in the early phase after infection of four rhesus macaques with pathogenic SIV-mac239. To determine transcription of interleukin (IL)-2, interferon (IFN)-gamma, IL-4, IL-6, and IL 10 we established a semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). After inoculation with SIV, all monkeys became productively infected and developed an acquired immunodeficiency syndrome (AIDS) like disease. Infection was associated with a proliferation dysfunction of monkey lymphocytes in response to PHA. In addition, a decreasing overall cytokine transcription could be observed during the course of SIV infection. These findings demonstrate that an impairment of the lymphocyte function is associated with a reduced cytokine transcription in the early phase of an immunodeficiency virus infection. The observed differences of cytokine expression might contribute to the impaired immune response of SIV-infected monkeys and HIV-infected humans. PMID- 9210281 TI - Simultaneous detection of hepatitis C virus and human immunodeficiency virus RNA in serum using amplicor PCR tests. AB - Several tests are currently available to assist in the diagnosis of the hepatitis C virus (HCV) and human immunodeficiency virus (HIV). Tests that actually detect or quantify these viruses are based on the polymerase chain reaction (PCR) technique. However, the application of PCR is limited by the cost, labor, time consumption, and potential for contamination. In this article we describe some procedures developed to reduce these limitations. We have developed and validated simultaneous detection methods for HIV RNA and HCV RNA in single serum samples using Amplicor PCR tests. The sensitivity and specificity of this method are comparable with the results obtained with commercial reverse transcription polymerase chain reaction (RT-PCR) techniques for HIV and HCV RNA detection. In addition we have modified the HIV Amplicor test for the RT-PCR procedure and the Chomczynski's method of RNA isolation. We hope that our method can find same applications in HIV and HCV coinfection research, blood screening, and medical diagnosis. PMID- 9210282 TI - Cell-mediated immunity to porcine reproductive and respiratory syndrome virus in swine. AB - Cell-mediated immunity has been demonstrated to be a necessary component of immunity against viral infection. Methods to detect T-cell mediated immune responses to porcine reproductive and respiratory syndrome virus (PRRSV) infection were established both in vitro as lymphocyte proliferation and in vivo as delayed-type hypersensitivity response (DTH). Optimal conditions for detection of lymphocyte proliferation were determined by testing different antigen concentrations and various stimulation periods. The proliferation response to PRRSV was antigen-specific and dose-dependent. The kinetics of the T-cell proliferation response to PRRSV were analyzed after primary and secondary exposure to virus. Lymphocyte proliferation was first detected at four weeks post infection (PI), peaked at 7 weeks PI, and declined after 11 weeks PI. The secondary response increased in magnitude. Experiments with blocking antibodies to porcine leukocyte antigens demonstrated that CD4+ T-cells were the major effector cells in the proliferation response. The in vivo response to PRRSV was shown by detection of a dose-dependent DTH reaction in infected pigs after intradermal challenge with UV-inactivated virus. These results demonstrate that pigs generate specific T-cell responses on PRRSV infection and provide a foundation for studying their role in protection. PMID- 9210283 TI - Apoptosis induction by human immunodeficiency virus type 1 (HIV-1) gp120 peptides. AB - We investigated the role of some gp120 peptides on the apoptosis induction in malignant T cell lines. We took advantage of recent findings reporting that three major regions of gp120 are important for CD4 binding. They consist of residues 256-262 in the C2 domain, residues 368-389 in the C3 domain, and residues 421-457 in C4 domain. We used a peptide from C2 domain (aa 250-263) the homologous major histocompatibility complex (MHC) class II peptide (aa 135-155) and three peptides from domain C4 (aa 414-434; 419-430; 428-445). We selected for this study the following human cell lines: CEM and Jurkat, two lymphoblastoid CD4-positive T cell line and U937, a myelomonocytic CD4 positive cell line. We demonstrated that the CD4-positive T cell lines, in the presence of gp120 250-263 peptide and DR 135-155 peptide, can be induced to accelerate apoptosis, while no effect in apoptosis induction was observed in the presence of 414-424 gp120 peptide. Interestingly, we have shown by fluorescence study, that the small sequence 414 419 must be responsible for the inhibition of binding of gp120 to the CD4 molecule. Indeed while 414-424 gp120 peptide is very efficient in CD4-gp120 binding inhibition, no effect is observed in the presence of either 419-430 or 428-445 peptide. PMID- 9210284 TI - Different affinity of monoclonal antibodies for conserved neutralizing epitopes on two strains of rubella virus. AB - There is, apparently, only one serological type of rubella virus (RV) in the population, although several isolates exist with different characteristics. Some authors failed to detect significant differences among RV strains by neutralization, hemagglutination inhibition, and enzyme immunoassay using polyclonal and monoclonal antibodies, but differences in growth, plaque morphology, and temperature sensitivity between vaccine and wild-type strains were shown by Chantler et al. (3) With the purpose of analyzing the possible differences among several strains of RV, we studied the affinity constant of two monoclonal antibodies (MAbs) for two conserved neutralizing epitopes. Wild-type Cordoba (regional isolation of a post-natal infection) and RA 27/3 (vaccine) strains of RV were tested. H3 and H14 MAbs were generated against wild-type Cordoba strain. They defined two epitopes with conserved neutralizing and hemagglutinating activity on both strains. The affinity of the MAbs (expressed as the affinity constant), was greater for Cordoba strain than for RA 27/3. Analyzing the results obtained, we conclude that the neutralizing epitopes defined by our MAbs on E1 glycoprotein are conserved in the two strains, but react with significative different affinities. This could be a way to characterize antigenically different viral strains of the same serotype. PMID- 9210285 TI - Immune responses in goats to caprine arthritis-encephalitis virus. AB - In this study, goats were experimentally infected with caprine arthritis encephalitis virus (CAEV). Anti-CAEV antibodies were detected in vivo using indirect enzyme-linked immunosorbent assay (ELISA) and Western blot assays. The results showed that the sensitivity of detection of anti-CAEV antibodies using indirect ELISA was relatively high, whereas Western blotting was very sensitive for detection of P28 anti-CAEV antibody as 4 days postinfection. Apparently, because of high immunogenicity of P28 antigen, the detection of antibodies specific for this viral protein may represent a valuable assay to detect early infection. CAEV related to other lentiviruses may be useful in studies of human immunodeficiency virus (HIV) and related viruses in animal models. PMID- 9210286 TI - Succinate: quinone oxidoreductases. Variations on a conserved theme. PMID- 9210287 TI - Phosphorous metabolites in boar spermatozoa. Identification of AMP by multinuclear magnetic resonance. AB - Boar spermatozoa revealed three prominent resonances in the 31P-NMR spectrum of intact cells. Two of these are known to be GPC and Pi, the third is a phosphomononoester (PME), the identification of which was carried out by proton detected 2D 1H,31P and 1H,13C chemical shift correlation experiments with gradient selection. The PME was unambiguously assigned to adenosine 5' monophosphate (AMP). The identification was confirmed by an AMP consuming enzymatic assay. Other physiologically relevant PME's, in particular inosine 5 monophosphate (IMP) and sugar phosphates, were excluded. The intensity of the 31P signal of AMP in boar sperm extract was much higher than those of ADP and ATP, and in intact cells only AMP but no ATP was visible. PMID- 9210288 TI - Anti-HIV active naphthyl analogues of HEPT and DABO. AB - 5-Ethyl-6-(1-naphthylmethyl)uracil and 5-ethyl-6-(1-naphthylmethyl)-2-thiouracil were alkylated to give, respectively. N-1 and S2, ethoxymethyl and methylthiomethyl uracil derivatives. 5-Ethyl-6-(1-naphthylmethyl)-2-thiouracil was also S2 alkylated with methyl bromoacetate. The products showed activity against HIV-1, and the N-1 alkylated derivatives were indeed highly active. PMID- 9210289 TI - Juvenile hormone in adult eusocial Hymenoptera: gonadotropin and behavioral pacemaker. AB - Studies on the role of juvenile hormone (JH) in adult social Hymenoptera have focused on the regulation of two fundamental aspects of colony organization: reproductive division of labor between queens and workers and age-related division of labor among workers. JH acts as a gonadotropin in the primitively eusocial wasp and bumble bee species studied, and may also play this role in the advanced eusocial fire ants. However, there is no evidence that JH acts as a traditional gonadotropin in the advanced eusocial honey bee or in the few other ant species that have recently begun to be studied. The role of JH in age-related division of labor has been most thoroughly examined in honey bees. Results of these studies demonstrate that JH acts as a "behavioral pacemaker," influencing how fast a worker grows up and makes the transition from nest activities to foraging. Hypotheses concerning the evolutionary relationship between the two functions of JH in adult eusocial Hymenoptera are discussed. PMID- 9210290 TI - Iron supplementation moderates but does not cure the Belgrade anemia. AB - Belgrade rats inherit microcytic, hypochromic anemia as an autosomal recessive trait (gene symbol b). Erythrocytes and tissue are iron deficient in the face of elevated TIBC (total iron binding capacity) and percent iron saturation; iron injections increased the number of erythrocytes but their appearance remained abnormal. We have investigated iron supplements to improve husbandry of b/b rats and to learn more about the underlying defect and its tissue distribution. Weekly i.m. (intramuscular) injections of iron-dextran (Imferon at 30 mg kg-1) improved the anemia but did not alter the red cell morphology. Certain diets also improved the health of b/b rats when compared to standard rat chows by the criteria of weight, survival to adulthood, hematology and reproduction. The critical nutritional factor turned out to be iron bioavailability, with ferrous iron added to the diet improving the health of Belgrade rats without affecting the underlying erythroid defect. Tissue iron measurements after dietary or parenteral supplementation confirmed the iron deficient status of untreated b/b rats and established that dietary ferrous iron partially relieved this deficiency, with injections leading to greater amounts of tissue iron. Serum iron and TIBC were also found to be elevated in untreated b/b rats, with dietary supplementation decreasing but not eliminating the elevation in TIBC. These studies indicate that iron supplements can improve the health of b/b rats without altering the underlying defect and also suggest that the mutation could alter iron uptake in the GI (gastrointestinal) tract. PMID- 9210291 TI - Tolerance to mercury chloride in Scenedesmus strains. AB - Mercury chloride toxicity was investigated in two strains of Chlorella and in a strain of Scenedesmus isolated from polluted areas in Tuscany (Italy). No Hg resistance was found in the autotrophic microorganisms isolated, but Scenedesmus sp. strain AR-2489, isolated from the Arno river, was able to grow at concentrations of up to 5 micrograms ml-1 of Hg. This concentration was twice that which inhibited growth of the two Chlorella strains and Scenedesmus acutus 8M, the reference strain from a culture collection. Photosynthesizing cells of Scenedesmus sp. AR-2489 showed reduced Hg uptake, with the highest percentage of Hg removal from the medium. Loss of Hg was not due to Hg(0) volatilization, as shown by a comparison test with the broad-spectrum Hg-resistant Pseudomonas putida FB1. The metabolic differences between Scenedesmus sp. strain AR-2489 and Siacatus strain 8M were: (1) higher growth rate (doubling time of 6.0 h versus 10.6 h); (2) higher O2 production rate (maximum 2 mumol h-1 mg-1 dry weight); and (3) higher intracellular pH during growth. The latter was imaged with a green fluorescence molecular probe (BCEFC-AM) and observed by scanning confocal laser microscopy (SCLM). The distribution of red-autofluorescence chlorophyll-a showed that strain AR-2489 had a rougher and hence more extended specific chloroplast surface than strain 8M. Hg tolerance in strain AR-2489 was related to the rapid increase in dissolved O2 in the medium and in intracellular pH; this caused a loss of soluble mercury transformed to insoluble mercury hydroxide, which is thermodynamically more stable at alkaline pH in highly oxygenated systems. PMID- 9210292 TI - Siderophore activity of chemically synthesized dihydroxybenzoyl derivatives of spermidines and cystamide. AB - Chemically synthesized dihydroxybenzoyl derivatives of spermidine and cystamide containing two-, three- and four-bidentates with the hydroxyl groups in 2,3 or 3,4 position were examined in cross-feeding tests using Gram-negative siderophore indicator strains carrying different iron-related markers, and two Mycobacterium spp. The catecholates were unable to feed tonB mutants of E. coli and S. typhimurium as well as the fepA, fiu, cir mutant of E. coli, pointing to a tonB- and fepA, cir, fiu-dependent transport. Bis(2,3-dihydroxybenzoyl)derivatives promoted Salmonella spp, E. coli, K. pneumoniae and P. aeruginosa strains significantly better than did 3,4-dihydroxybenzoyl derivatives. N4-substituted spermidines acted more effectively than non-substituted derivatives. Bis(2,3 dihydroxybenzoyl) cystamide was superior to the other catecholates tested in growth promotion of Gram-negative bacteria. The two four-bidentates and the tri bidentate reacted to K. pneumoniae in an inhibitory mode. The position of the hydroxyl groups did not significantly influence the growth promotion of M. smegmatis and M. fortiutum in the cases of substituted spermidines and of cystamides. PMID- 9210293 TI - Hydroxyl free radicals generated by vanadyl[IV] induce cell blebbing in mitotic human Chang liver cells. AB - Vanadium has recently been reported to induce interphase and M-phase (mitotic) programmed cell death via the generation of hydroxyl free radicals (OH*). In this paper, the effects of antioxidants on: (a) vanadyl[IV]-generated OH* free radical levels; and (b) cellular glutathione in vanadyl [IV]-treated Chang liver cells were evaluated. The surface morphology of vanadyl-treated mitotic cells was studied by confocal and scanning microscopy. The free radical scavengers zinc chloride, glucose and thiourea reduced the levels of vanadyl-induced OH* free radicals and partially prevented the depletion of cellular glutathione. Concurrent with OH* free radical production, vanadyl-treated telophase cells exhibited excessive cell blebbing and cell shrinkage. The morphological features demonstrated in vanadyl-induced mitotic programmed cell death as a consequence of oxidative stress is novel. PMID- 9210294 TI - A study on ascorbate inhibition of ceruloplasmin ferroxidase activity. AB - The ferroxidase activity of ceruloplasmin is often determined according to the method of Johnson et al. (1967), using apotransferrin for trapping ferric ions generated by the enzyme; spectrophotometrically monitoring the Fe(3+)-transferrin formation at pH 6.0. Reports have shown that ascorbate inhibits this reaction, and it is hypothesized that the effect could be of physiological significance in individuals with a high ascorbate to ceruloplasmin ratio in plasma (e.g. premature babies). The present study shows that the inhibitory effect of ascorbate rapidly decreases with increasing pH. At pH 7.4 no significant effect was observed, the result suggesting that ascorbate is not a physiological inhibitor of ceruloplasmin. Furthermore, experiments demonstrate that at acidic pH the inhibitory effect of ascorbate on the rate of Fe(3+)-transferrin formation is not primarily due to an interaction with ceruloplasmin, but to a reduction of enzymically generated ferric ions before they are bound to apotransferrin. PMID- 9210295 TI - Proliferative and morphological changes induced by vanadium compounds on Swiss 3T3 fibroblasts. AB - Vanadium compounds are shown to have a mitogenic effect on fibroblast cells. The effects of vanadate, vanadyl and pervanadate on the proliferation and morphological changes of Swiss 3T3 cells in culture are compared. Vanadium derivatives induced cell proliferation in a biphasic manner, with a toxic-like effect at doses over 50 microM, after 24 h of incubation. Vanadyl and vanadate were equally potent at 2.5-10 microM. At 50 microM vanadate inhibited cell proliferation, whereas slight inhibition was observed at 100 microM of vanadyl. At 10 microM pervanadate was as potent as vanadate and vanadyl in stimulating fibroblast proliferation, but no effect was observed at lower concentrations. A pronounced cytotoxic-like effect was induced by pervanadate at 50 microM. All of these effects were accompanied by morphological changes: transformation of fibroblast shape from polygonal to fusiform; retraction with cytoplasm condensation; and loss of lamellar processes. The magnitude of these transformations correlates with the potency of vanadium derivatives to induce a cytotoxic-like effect: pervanadate > vanadate > vanadyl. These data suggest that the oxidation state and coordination geometry of vanadium determine the degree of the cytotoxicity. PMID- 9210296 TI - Hypothesis: iron chelation plays a vital role in neutrophilic inflammation. AB - Neutrophil influx into tissues occurs in many diverse diseases and can be associated with both beneficial and injurious effects. We hypothesize that the stimulus for certain neutrophilic inflammatory responses can be reduced to a series of competing reactions for iron, with either a labile or reactive coordination site available, between host chelators and chelators not indigenous to that specific living system. The iron focuses the transport of host phagocytic cells through a metal catalyzed generation of oxidant sensitive mediators including cytokines and eicosanoids. Many of these products are chemotactic for neutrophils. We also postulate that the iron increases the activity of the phagocyte associated NADPH oxidoreductase in the neutrophil. The function of this enzyme is likely to be the generation of superoxide in the host's attempt to chemically reduce and dislodge the iron from its chelate complex. After the reoxidation of Fe2+ in an aerobic environment, Fe3+ will be coordinated by host lactoferrin released by the neutrophil. When complexed by this glycoprotein, the metal does not readily undergo oxidation/reduction and is safely transported to the macrophages of the reticuloendothelial system where it is stored in ferritin. Finally, we propose that the neutrophil will attempt to destroy the chelator not indigenous to the host by releasing granular contents other than lactoferrin. Inability to eliminate the chelator allows this sequence to repeat itself, which can lead to tissue injury. Such persistence of a metal chelate in the host may be associated with biomineralization, fibrosis, and cancer. PMID- 9210297 TI - 1H NMR analysis of novel sialylated and fucosylated lactose-based oligosaccharides having linear GlcNAc(beta 1-6) Gal and Neu5Ac(alpha 2-6) GlcNAc sequences. AB - Three novel oligosaccharides of human infant faeces have been fully characterised by methylation analysis and 500/600 MHz 1H NMR spectroscopy including DQF-COSY, TQF-COSY, TOCSY and ROESY experiments. The oligosaccharides were shown to be lactose-based structures two of which were substituted at C-6 of Gal with either the Le(x) trisaccharide, Gal(beta 1-4)[Fuc(alpha 1-3)]GlcNAc(beta 1-, or Neu5Ac(alpha 2-6)Gal(beta 1-4)GlcNAc(beta 1-. They differ from other free oligosaccharides previously isolated from the human by having the (1-->6) linkage to Gal in the absence of a (1-->3) branch. The third oligosaccharide has Neu5Ac(alpha 2-6) linked to GlcNAc of the trisaccharide GlcNAc(beta 1-3)Gal(beta 1-4)Glc. This is a linear fragment of the disialylated tetrasaccharide sequence Neu5Ac(alpha 2-3)Gal(beta 1-3)[Neu5Ac(alpha 2-6)]GlcNAc(beta 1-found in the milk oligosaccharide disialyl LNT (the GlcNAc residue of the tetrasaccharide linked to lactose) and also of N-linked chains (GlcNAc linked to Man). PMID- 9210298 TI - Chondroitin ABC lyase digestion of an ascidian dermatan sulfate. Occurrence of unusual 6-O-sulfo-2-acetamido-2-deoxy-3-O-(2-O-sulfo-alpha-L-idopyranosyluronic acid)-beta-D-galactose units. AB - A dermatan sulfate-like glycosaminoglycan was isolated from the body of the ascidian Ascidia nigra (J. Biol. Chem. 270: 31027-31036, 1995). 1H NMR and fast atom bombardment mass spectrometry (FAB-MS) spectra of the tetra and disaccharides formed by chondroitin ABC lyase digestion support the proposed repeating disaccharide structure for this glycosaminoglycan, [-->4)-alpha-L IdoA(2SO4)-(1-->3)-beta-D-GalNAc(6SO4)-(1-->] , which differ from mammalian dermatan sulfate in its sulfation at both 2-position of the alpha-L-iduronic acid and the 6-position of the N-acetyl-beta-D-galactosamine residue. PMID- 9210299 TI - The structure of the exopolysaccharide of Pseudomonas fluorescens strain H13. AB - An acidic exopolysaccharide was isolated from P. fluorescens strain H13. The structure of the polysaccharide repeating unit was determined using chemical methods and 1D and 2D NMR techniques. The repeating unit was characterized as a trisaccharide composed of D-glucose, 2-acetamido-2-deoxy-D-glucose and 4-O-acetyl 2-acetamido-2-deoxy-D-mannuronic acid. PMID- 9210300 TI - Synthesis of methyl alpha-glycosides of some higher oligosaccharide fragments of the O-antigen of Vibrio cholerae O1, serotype Inaba and Ogawa. AB - The title oligosaccharides, the tri- through the hexasaccharide in the Inaba series and the penta- and the hexasaccharide in the Ogawa series, have been synthesized using 1-thioglycosides of precursors to 3-O-benzyl-perosamine (4 amino-4,6-dideoxy-D-mannose) as building blocks and N-iodosuccinimide/silver triflate as a promoter. The azido groups in the assembled oligosaccharides were reduced to amino groups, which were then acylated using 2,4-O-benzylidene-3-deoxy L-glycero-tetronic acid as the derivatizing reagent. Catalytic hydrogenolysis, simultaneously of the benzyl and benzylidene groups, gave the desired products that were characterized by 1H and 13C NMR spectroscopy. PMID- 9210301 TI - Synthesis and glycosidic coupling reaction of substituted 2,6 dioxabicyclo[3.1.0]hexanes: 1,2-anhydro-3,5-di-O-benzyl-alpha-D-ribofuranose. PMID- 9210302 TI - Effects of intermittent punishment on self-injurious behavior: an evaluation of schedule thinning. AB - Although the use of punishment often raises ethical issues, such procedures may be needed when the reinforcers that maintain behavior cannot be identified or controlled, or when competing reinforcers cannot be found. Results of several studies on the effects of intermittent schedules of punishment suggest that therapists must use fairly rich schedules of punishment to suppress problem behavior. However, residential caretakers, teachers, and parents often have difficulty implementing programs that require constant monitoring of the client's behavior. In this study, we examined the feasibility of gradually thinning the delivery of punishment from a continuous schedule to an intermittent schedule during the course of treatment for self-injurious behavior (SIB). Results of functional analyses for 5 individuals who had been diagnosed with profound mental retardation indicated that their SIB was not maintained by social consequences. Treatment with continuous schedules of time-out (for 1 participant) or contingent restraint (for the other 4 participants) produced substantial reductions in SIB. When they were exposed to intermittent schedules of punishment (fixed-interval [FI] 120 s or FI 300 s), SIB for all but 1 of the participants increased to levels similar to those observed during baseline. For these 4 participants, the schedule of punishment was gradually thinned from continuous to FI 120 s or FI 300 s. For 2 participants, SIB remained low across the schedule changes, demonstrating the utility of thinning from continuous to intermittent schedules of punishment. Results for the other 2 participants showed that intermittent punishment was ineffective, despite repeated attempts to thin the schedule. PMID- 9210303 TI - Noncontingent presentation of attention and alternative stimuli in the treatment of attention-maintained destructive behavior. AB - Previous research has demonstrated that destructive behavior may be reduced through noncontingent presentation of attention when attention is identified as the stimulus responsible for behavioral maintenance. Because it may not always be possible to deliver attention in all situations, we examined the extent to which alternative stimuli that have been identified through a choice assessment would substitute for attention (the functional analysis-based reinforcer) in a noncontingent reinforcement procedure. Prior to treatment, functional analyses demonstrated that the destructive behavior of 2 clients with mental retardation was maintained by adult attention. Next, a stimulus choice assessment identified highly preferred tangible items for the 2 clients. Finally, we compared the effectiveness of two noncontingent reinforcement procedures: continuous noncontingent access to attention and continuous noncontingent access to the tangible item identified in the choice assessment. For both clients, these noncontingent reinforcement procedures reduced destructive behavior. The results are discussed in terms of the clinical implications for the treatment of destructive behavior using functional and alternative stimuli. PMID- 9210304 TI - Noncontingent delivery of arbitrary reinforcers as treatment for self-injurious behavior. AB - Results of recent research have shown that noncontingent reinforcement (NCR) can be effective in reducing the frequency of behavior problems. In typical NCR applications, the reinforcer that is responsible for behavioral maintenance (as demonstrated through a functional analysis) no longer follows occurrences of the target behavior but instead is delivered according to a time-based schedule. Thus, it is unclear if NCR would be effective if the target behavior continued to be reinforced or if arbitrary reinforcers (i.e., those irrelevant to behavioral maintenance) were substituted for the maintaining reinforcers in the NCR procedure. In this study, 2 individuals whose self-injurious behavior (SIB) was maintained by positive reinforcement were exposed to conditions in which arbitrary and maintaining reinforcers were withheld and were delivered either contingently or noncontingently. Results indicated that noncontingent delivery of arbitrary reinforcers was effective in reducing SIB even though occurrences of SIB produced access to the maintaining reinforcer. These results suggest that (a) arbitrary reinforcers may sometimes be substituted for maintaining reinforcers, (b) an important component of NCR procedures is alteration of a behavior's establishing operation, and (c) NCR with arbitrary reinforcers might therefore be effective when maintaining reinforcers cannot be identified or withheld during the course of treatment. PMID- 9210305 TI - On the relation of mands and the function of destructive behavior. AB - When standard analogue functional analysis procedures produce inconclusive results in children with conversational speech, the child's mands may help to identify the function of destructive behavior. In the current investigation, functional analyses conducted with 2 children who exhibited self-injury, aggression, and property destruction were undifferentiated across conditions. Based on informal observations and school and parental report, an analysis was conducted using mands to help determine the function of the destructive behavior. Using a multielement design, the therapist's compliance with the child's mands occurred either on a fixed-ratio (FR) 1 schedule or contingent on destructive behavior. Destructive behavior occurred at high and consistent levels when reinforcement of mands was contingent on destructive behavior and at near-zero levels when reinforcement of mands occurred on the FR 1 schedule. Based on these results, a second analysis was conducted in which compliance to mands occurred only when the child appropriately requested it (i.e., functional communication training plus extinction) and, for 1 child, compliance with mands was terminated contingent upon destructive behavior (i.e., functional communication training plus response cost). For both children, the rates of destructive behavior decreased markedly. The results suggest that assessing the child's mands may be useful in decreasing destructive behavior when a functional analysis is inconclusive. PMID- 9210306 TI - Evaluation of the "control over reinforcement" component in functional communication training. AB - The effectiveness of functional communication training (FCT) as a treatment for behavior disorders has been attributed to a number of variables, one of which is the individual's ability to exert control over the delivery of reinforcement. We evaluated this component of FCT by exposing individuals to conditions in which their behavior either did or did not affect the delivery of reinforcement. Three adults with mental retardation who engaged in self-injurious behavior (SIB) participated. Following a functional analysis of their SIB, the effects of FCT were compared to those of noncontingent reinforcement (NCR) in a multielement design. The amount of reinforcement during both conditions was equated by yoking the schedule of reinforcement during NCR sessions to that in effect during FCT sessions. Results indicated that FCT and NCR were equally effective in reducing the SIB of all participants and suggest that control over reinforcement delivery may not affect the degree to which FCT produces behavioral suppression. However, a different benefit of FCT was evident in the results: More consistent increases in the alternative response were observed during the FCT condition than during the NCR condition. PMID- 9210307 TI - The use of positive and negative reinforcement in the treatment of escape maintained destructive behavior. AB - We identified 3 clients whose destructive behavior was sensitive to negative reinforcement (break from tasks) and positive reinforcement (access to tangible items, attention, or both). In an instructional context, we then evaluated the effects of reinforcing compliance with one, two, or all of these consequences (a break, tangible items, attention) when destructive behavior produced a break and when it did not (escape extinction). For 2 clients, destructive behavior decreased and compliance increased when compliance produced access to tangible items, even though destructive behavior resulted in a break. For 1 client, extinction was necessary to reduce destructive behavior and to increase compliance. Subsequently, when the schedule of reinforcement for compliance was faded for all clients, destructive behavior was lower and fading proceeded more rapidly when compliance produced multiple functional reinforcers (i.e., a break plus tangible items or attention) and destructive behavior was on extinction. The results are discussed in terms of the effects of relative reinforcement value and extinction on concurrent operants. PMID- 9210308 TI - Identifying instructional tasks that occasion problem behaviors and assessing the effects of student versus teacher choice among these tasks. AB - Levels of problem behavior were assessed when 4 students with severe disabilities received instruction on preferred versus nonpreferred tasks and when tasks of each type were chosen by the teacher rather than by the student. In Phase 1, interview and direct observation assessments were conducted to identify relative preferences for academic tasks. In Phase 2, the effects of these lower preference and higher preference tasks on the rate of problem behavior were evaluated using a multielement design. The results showed that lower preference tasks were associated with higher rates of problem behaviors and that students, when given a choice, consistently selected the tasks that had been identified through interview and direct observation as higher preference. In Phase 3, we assessed whether allowing the students to choose between pairs of lower preference tasks or between pairs of higher preference tasks reduced problem behavior relative to a condition in which the teacher selected the same tasks. For 2 of 4 students, the rates of problem behavior were lower when students (rather than the teacher) selected the lower preference activity. Higher preference tasks for 3 students were associated with relatively low rates of problem behavior regardless of whether the student or the teacher selected the task. PMID- 9210309 TI - Toward the development of structured criteria for interpretation of functional analysis data. AB - Using functional analysis results to prescribe treatments is the preferred method for developing behavioral interventions. Little is known, however, about the reliability and validity of visual inspection for the interpretation of functional analysis data. The purpose of this investigation was to develop a set of structured criteria for visual inspection of multielement functional analyses that, when applied correctly, would increase interrater agreement and agreement with interpretations reached by expert consensus. In Study 1, 3 predoctoral interns interpreted functional analysis graphs, and interrater agreement was low (M = .46). In Study 2, 64 functional analysis graphs were interpreted by a panel of experts, and then a set of structured criteria were developed that yielded interpretive results similar to those of the panel (exact agreement = .94). In Study 3, the 3 predoctoral interns from Study 1 were trained to use the structured criteria, and the mean interrater agreement coefficient increased to .81. The results suggest that (a) the interpretation of functional analysis data may be less reliable than is generally assumed, (b) decision-making rules used by experts in the interpretation of functional analysis data can be operationalized, and (c) individuals can be trained to apply these rules accurately to increase interrater agreement. Potential uses of the criteria are discussed. PMID- 9210310 TI - Effects of high-preference single-digit mathematics problem completion on multiple-digit mathematics problem performance. AB - The purpose of this study was to examine the effects of a sequence of three single-digit (1 digit x 1 digit) multiplication problems on the latency to initiate multiple-digit (3 digit x 3 digit) multiplication problems for 2 students in an alternative education school. Data showed that (a) during the preference assessment, both students selected the single-digit problems in a majority of the sessions, and (b) intervention resulted in a decrease in latency between problems for both students. Results are discussed in relation to using high-preference sequences to promote behavioral momentum in academic content areas. PMID- 9210311 TI - Picture naming, matching to sample, and head injury: a stimulus control analysis. AB - Computer-based procedures were used to examine oral naming and matching-to-sample performances in an adult with a head injury. Relatively few errors occurred when pictures were (a) named, (b) matched to dictated names presented simultaneously, (c) matched to dictation after a delay, and (d) matched to identical pictures presented simultaneously. More errors occurred on delayed than on simultaneous identity matching. On delayed matching trials, fewer errors occurred when instructions to name the samples were given. PMID- 9210313 TI - Purification of p-nitrobenzyl C-functionalized diethylenetriamine pentaacetic acids for clinical applications using anion-exchange chromatography. AB - A general process for the purification of large quantities (5-10 g) of p nitrobenzyl C-functionalized diethylenetriamine pentaacetic acids is reported. The method of choice to achieve purification for clinical applications is anion exchange chromatography. PMID- 9210314 TI - Peptide separation by pH-zone-refining countercurrent chromatography. AB - Peptides without protecting groups have been successfully separated by pH-zone refining countercurrent chromatography (CCC) using an ion-pair reagent, di-(2 ethylhexyl)phosphoric acid (DEHPA), as a modifier in the stationary phase. Preliminary studies indicated that two parameters, i.e., the DEHPA concentration in the stationary phase and hydrophobicity of the solvent system should be adjusted according to the hydrophobicity of the analytes. Hydrophobic and hydrophilic groups of dipeptides were each separated under optimized conditions. The method was successfully applied to gram-quantity separations of bacitracin complex and bovine insulin. PMID- 9210312 TI - Antecedent influences on behavior disorders. AB - The influence of antecedent events on behavior disorders has been relatively understudied by applied behavior analysts. This lack of research may be due to a focus on consequences as determinants of behavior and a historical disagreement on a conceptual framework for describing and interpreting antecedent variables. We suggest that antecedent influences can be described using terms derived from basic behavioral principles and that their functional properties can be adequately interpreted as discriminative and establishing operations. A set of studies on assessment and treatment of behavior disorders was selected for review based on their relevance to the topic of antecedent events. These studies were categorized as focusing on assessment of antecedent events, antecedent treatments for behavior disorders maintained by either positive or negative reinforcement, and special cases of antecedent events in behavior disorders. Some directions for future research on antecedent influences in the analysis and treatment of behavior disorders are discussed. PMID- 9210315 TI - Drug quantitation on a benchtop liquid chromatography-tandem mass spectrometry system. AB - The specificity and selectivity of LC-MS-MS is illustrated to explain why LC-MS MS has become the method of choice for quantitation within the pharmaceutical industry. Two assays are described that demonstrate the facility with which new ion trap technology can utilize the selectivity and sensitivity of LC-MS-MS to quantitate trace level components within complex matrices, in particular human plasma. One assay undergoes a validation procedure and demonstrates the utility of this new technology for drug quantitation within a regulated environment. PMID- 9210316 TI - Separation of extracts from biological tissues into polycyclic aromatic hydrocarbon, polychlorinated biphenyl and polychlorinated dibenzo-p dioxin/polychlorinated dibenzofuran fractions prior to analysis. AB - A low-pressure liquid chromatography method is presented for separating polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs) and polychlorinated dibenzo-p-dioxins/polychlorinated dibenzofurans (PCDDs/PCDFs) from biological tissue extracts. After removing lipid from extracts, the PAHs are separated from PCBs and PCDDs/PCDFs on a deactivated 13-24 microns silica gel column. The PCBs are subsequently separated from PCDDs/PCDFs by collecting the first fraction from an automated three column cleanup procedure for PCDDs/PCDFs. The complete method has been used to obtain high recoveries of the three compound classes for analysis by GC-electron capture detection (PCBs) or GC-MS (PAHs and PCDDs/PCDFs). PMID- 9210317 TI - Analysis of DNA fragmentation using a dynamic size-sieving polymer solution in capillary electrophoresis. AB - Various natural and induced processes cause DNA fragmentation. Examples of these processes include apoptosis, enzymatic digestion, free radical production from ionizing radiation, photoscission by laser radiation and thermal degradation. Slab gel electrophoresis has been used most often to monitor such DNA damage. We have investigated with capillary electrophoresis the use of a new size-sieving polymer solution, TreviSol-CE (TS-CE), to monitor the DNA fragments produced from a variety of degradation processes. This polymer solution provides high run-to run migration time and peak width reproducibilities and high separation efficiency of double-stranded DNA fragments in the 500 to 7000 base pair size range. Analysis of apoptotic DNA fragments suggested the presence of multiple nucleosomes within each cell type investigated. For irradiated DNA standards, peak-width-at-half-height and peak area were used to monitor the progress of DNA fragmentation. For both apoptotic DNA and irradiated DNA standards, fine structural features of fragmentation were revealed. PMID- 9210318 TI - Photophysical and photobiological processes in the photodynamic therapy of tumours. AB - Photodynamic therapy (PDT) is an innovative and attractive modality for the treatment of small and superficial tumours. PDT, as a multimodality treatment procedure, requires both a selective photosensitizer and a powerful light source which matches the absorption spectrum of the photosensitizer. Quadra Logic's Photofrin, a purified haematoporphyrin derivative, is so far the only sensitizer approved for phase III and IV clinical trials. The major drawbacks of this product are the lack of chemical homogeneity and stability, skin phototoxicity, unfavourable physicochemical properties and low selectivity with regard to uptake and retention by tumour vs. normal cells. Second-generation photosensitizers, including the phthalocyanines, show an increased photodynamic efficiency in the treatment of animal tumours and reduced phototoxic side effects. At the time of writing of this article, there were more than half a dozen new sensitizers in or about to start clinical trials. Most available data suggest a common mechanism of action. Following excitation of photosensitizers to long-lived excited singlet and/ or triplet states, the tumour is destroyed either by reactive singlet oxygen species (type II mechanism) and/or radical products (type I mechanism) generated in an energy transfer reaction. The major biological targets of the radicals produced and of singlet oxygen are well known today. Nucleic acids, enzymes and cellular membranes are rapidly attacked and cause the release of a wide variety of pathophysiologically highly reactive products, such as prostaglandins, thromboxanes and leukotrienes. Activation of the complement system and infiltration of immunologically active blood cells into the tumorous region enhance the damaging effect of these aggressive intermediates and ultimately initiate tumour necrosis. The purpose of this review article is to summarize the up-to-date knowledge on the mechanisms responsible for the induction of tumour necrotic reactions. PMID- 9210319 TI - N-conjugates of 2,5-disubstituted pyrrole and glutathione. Evaluation of their potency as antioxidants against photosensitization of NCTC 2544 keratinocytes by excess endogenous protoporphyrin IX. AB - A novel glutathione compound in which the amino group has been derivatized by a 2,5-dimethyl pyrrole is shown to be very effective against cell photosensitization in vitro. Protoporphyrin IX either added to the medium or produced endogenously by incubation of NCTC 2544 keratinocytes with 5 aminolevulinic acid has been chosen as the photosensitizer. The antioxidant effectiveness of glutathione-pyrrole derivatives against protoporphyrin photosensitization depends critically on the type of 2,5 substitution on the pyrrole ring. This structure-function relationship may be attributed to the difference in compartmentation and/or uptake of the various glutathione-pyrrole derivatives under study. The 2,5-dimethyl pyrrole derivative is much more effective than glutathione as a protective agent against phototoxic reactions induced by protoporphyrin IX. PMID- 9210320 TI - In vivo photodynamic therapy with the new near-IR absorbing water soluble photosensitizer lutetium texaphyrin and a high intensity pulsed light delivery system. AB - An in vivo fluorescence monitoring and photodynamic therapy (PDT) study was performed using the new photosensitizer lutetium texaphyrin (Lu-Tex). This photosensitizer is water soluble and has the additional advantage of strong absorption near 730 nm. C26 colon carcinoma was transplanted in the foot of BALB/c mice. In vivo fluorescence spectroscopy was applied to study Lu-Tex tissue distribution kinetics. For this purpose, fluorescence intensity both in the foot with the tumor and in the normal foot was measured in vivo by the laser-induced fluorescence (LIF) system. For PDT, both feet of the mice were irradiated simultaneously with the use of a new high intensity pulsed light delivery system, the Photodyne. The results of the LIF measurements showed that the maximal fluorescence intensity ratio between the normal and tumor bearing foot (FIR) was observed 24-48 h after the agent injection. Photoirradiation with doses from 90 to 240 J cm-2 (0.6 J cm-2 per 2 ms pulse, 1 Hz) 24 h after injection of Lu-Tex at a dose of 10 mg kg-1 caused significant tumor necrosis and delay in the tumor growth rate. The antitumor effect was enhanced with increasing light doses. Normal tissue response to PDT with Lu-Tex was determined as the damage index of the normal foot, which was irradiated simultaneously with the tumor bearing foot. The normal tissue response after PDT with Lu-Tex was compared with 5 aminolevulinic acid (ALA) induced protoporphyrin IX (PP), chlorin e6 (Chl) and Photofrin (PII) at the same values of antitumor effect. The results showed that at 50, 80 and 100% inhibition of tumor growth the orders of the values of normal foot damage indexes were as follows: ALA > Lu-Tex > or = PII > Chl, PII > ALA > Lu-Tex > Chl and PII > Lu-Tex > ALA > Chl respectively. PMID- 9210321 TI - A study of the binding of merocyanine 540 to myeloid leukemia M1 cells using an intensified charge-coupled device for fluorescence imaging microscopy. AB - The binding of merocyanine 540 (MC540) to murine myeloid leukemia (M1) cells and normal erythrocytes was measured by fluorescence digital imaging microscopy using an intensified charge-coupled device. It was found that, on average, about three times more MC540 were bound to a unit membrane area of M1 cells than erythrocytes, a result consistent with previous studies. However, it was shown for the first time that MC540 binding varied significantly from one M1 cell to the next, and about 15% of the sensitized M1 cells were as MC540-negative as normal erythrocytes. Using the leukemic inhibitory factor as a differentiation inducer, M1 cells were induced to differentiate into mature macrophage-like cells in vitro. Such treatment lowered the average MC540 binding by about one-third but did not affect the cell-to-cell variation significantly. PMID- 9210322 TI - Sites of preferred interaction between double-stranded pBR322 DNA and 7 methylpyrido[3,4-c]psoralen. AB - 7-Methylpyrido[3,4-c]psoralen has recently been found to photosensitize the crosslinking and cleavage of double-stranded pBR322 DNA (4361 bp) with an unusual degree of site specificity. Following cleavage of the photosensitized pBR322 with the restriction enzymes Pvu II and BamH I, the main fragments were analyzed electrophoretically. Only three possible cleavage areas on the plasmid were suggested by this analysis. They were within the ranges bounded by the positions 801-926, 3172-3303 and 4226-4310. The sensitizer also induced selective cleavage in the related plasmid pUC19 (2686 bp), but the pattern was somewhat more complicated and was not analyzed in detail. PMID- 9210323 TI - Photosensitizing activity of the anti-bacterial drugs sulfamethoxazole and trimethoprim. AB - The photochemical reactions in vitro of sulfamethoxazole alone and in combination with trimethoprim were studied to obtain information on the photosensitization mechanism. Sulfamethoxazole in aqueous solution, on exposure to UVB radiation, generates free radicals and singlet oxygen, with the neutral molecule being at least twice as active as the sulfamethoxazole anion. Photoexcited sulfamethoxazole can participate in electron transfer to cytochrome-c and nitro blue tetrazolium, and sensitizes the peroxidation of linoleic acid and the hemolysis of human erythrocytes, predominantly by a free radical mechanism. Trimethoprim is relatively inactive in the same photochemical systems. PMID- 9210325 TI - Pharmacokinetics of N-aspartyl chlorin e6 in cancer patients. AB - We examined the pharmacokinetics of the photosensitizer N-aspartyl chlorin e6 in a group of cancer patients. While the drug persisted in plasma for as long as six weeks, there was no evidence of fluorescent NPe6 metabolites during this interval. Kinetics of drug elimination from plasma were consistent with a 2 compartment model with half-lives of approximately 9 hr (57%) and 134 hr (43%). The drug was bound to plasma albumin+other heavy proteins (65%) > HDL (35%) > > LDL (1-2%). These relative values did not change for as long as 21 days after drug administration. The long persistence of NPe6 in plasma was not associated with extended skin photosensitization. PMID- 9210324 TI - The influence of the presence of lipid on the aggregation of 8,12-diethyl farnesyl bacteriochlorophyll c located in adsorbed layers and monolayers. AB - The photoacoustic spectra and time-resolved delayed luminescence spectra in the microsecond time range were measured for layers of 8,12-diethyl farnesyl bacteriochlorophyll c adsorbed on quartz supports by solvent evaporation and as Langmuir-Blodgett monolayers. Both types of model system were also investigated with the addition of lipid. The data showed a very strong influence of lipid addition on pigment aggregation. In samples with synthetic and natural lipid addition, the pigments were found to be predominantly in the monomeric and dimeric states, whereas in the same type of sample without lipid, the pigments were aggregated to a higher degree. The influence of the presence of lipid on the aggregation of bacteriochlorophyll c in monolayers and adsorbed layers may also suggest that the contact of various pigment molecules with the lipids surrounding the chlorosome may influence the formation of various pigment aggregates in vivo. The synthetic lipid L-alpha-phosphatidylcholine dipalmitoyl and the natural lipid L-alpha-phosphatidylcholine type IVS from soy beans were used. In the latter case, only adsorbed layers were investigated. Our interpretation is preliminary as only one 8,12-diethyl farnesyl bacteriochlorophyll c homologue was present in our systems. PMID- 9210326 TI - Overexpression of the small heat shock protein, hsp27, confers resistance to hyperthermia, but not to oxidative stress and UV-induced cell death, in a stably transfected squamous cell carcinoma cell line. AB - The 27 kD heat shock protein (hsp27) is expressed in human keratinocytes in association with differentiation in vitro and in situ. This study was conducted to investigate whether the expression of hsp27 in keratinocytes is associated with increased resistance to the deleterious effects of heat and UV radiation. A transfection vector carrying the human gene for hsp27, under the control of hsp27 as well as the SV40 promoter (pSG2711, M. Jaattela et al., EMBO J. 11 (1992) 3507 3512), was introduced together with a neomycin-resistance gene into the squamous cell carcinoma cell line A431. Cells were exposed to either UVA, UVB, head (45 degrees C, 4 h) or hydrogen peroxide (0.025-0.5 mM) and the percentage of surviving cells was determined. Overexpression of hsp27 induced increased resistance to hyperthermia, but not to hydrogen peroxide-mediated oxidative injury. When cells were exposed to increasing amounts of UVA (5-80 J cm-2) and UVB (4-64 mJ cm-2), the percentage of surviving cells was identical for clones overexpressing hsp27 and control clones. From these data, we conclude that hsp27 is a mediator of thermotolerance, but does not protect keratinocytes from UV induced cell death. PMID- 9210329 TI - Reduced gravitropism in hypocotyls of starch-deficient mutants of Arabidopsis. AB - Gravitropism was examined in dark- and light-grown hypocotyls of wild-type (WT), two reduced starch mutants (ACG 20 and ACG 27), and a starchless mutant (ACG 21) of Arabidopsis. In addition, the starch content of these four strains was studied with light and electron microscopy. Based on time course of curvature and orientation studies, the graviresponse in hypocotyls is proportional to the amount of starch in a genotype. Furthermore, starch mutations seem to primarily affect gravitropism rather than differential growth since both phototropic curvature and growth rates among the four genotypes are approximately equal. Our results suggest that gravity perception may require a greater plastid mass in hypocotyls compared to roots. The kinetics of gravitropic curvature also was compared following reorientation at 45 degrees, 90 degrees, and 135 degrees. As has been reported for other plant species, the optimal angle of reorientation is 135 degrees for WT Arabidopsis and the two reduced starch mutants, but the magnitude of curvature of the starchless mutant appears to be independent of the initial angle of displacement. Taken together, the results of the present study and our previous experiments with roots of the same four genotypes [Kiss et al. (1996) Physiol. Plant. 97: 237] support a plastid-based hypothesis for gravity perception in plants. PMID- 9210327 TI - Effects of abscisic acid and cytoplasmic pH on potassium and chloride efflux in Arabidopsis thaliana seedlings. AB - The effects of ABA, isobutyric acid (IBA) and nicotine on K+ and Cl- efflux were studied in Arabidopsis thaliana seedlings, and the role of pHcyt and Em in the regulation of the efflux of these ions was discussed. The data show that treatments with IBA and nicotine influenced in opposite directions the efflux of either K+ or Cl-: K+ efflux was increased by nicotine and reduced in the presence of IBA, whereas Cl- efflux was stimulated by IBA and decreased by nicotine treatment. Under all the conditions tested ABA induced cytoplasmic acidification and inhibition of K+ and Cl- net efflux. Experiments aimed to estimate the individual contribution of pHcyt and Em in modulating K+ efflux indicated that, within the range of acidic pHcyt values, a regulation of K+ efflux was imposed by pHcyt on the control exerted by Em, the efflux being inhibited by lower pHcyt values. Conversely, in the alkaline side of pHcyt K+ efflux seemed linked only to the Em values. These results are consistent with the hypothesis that the decrease in K+ efflux observed in non-stomatal tissues in the presence of ABA may be mediated by the cytoplasmic acidification induced by the hormone. PMID- 9210330 TI - Mutations in the SGR4, SGR5 and SGR6 loci of Arabidopsis thaliana alter the shoot gravitropism. AB - Shoots of higher plants grow upward in response to gravity. To elucidate the molecular mechanism of this response, we have isolated shoot gravitropism (sgr) mutants in Arabidopsis thaliana. In this report, we describe three novel mutants, sgr4-1, sgr5-1 and sgr6-1 whose inflorescence stems showed abnormal gravitropic responses as previously reported for sgr1, sgr2 and sgr3. These new sgr mutations were recessive and occurred at three independent genetic loci. The sgr4-1 mutant showed severe defect in gravitropism of both inflorescence stem and hypocotyl but were normal in root gravitropism as were sgr1 and sgr2. The sgr5-1 and sgr6-1 mutants showed reduced gravitropism only in inflorescence stems but normal in both hypocotyls and roots as sgr3. These results support the hypothesis that some mechanisms of gravitropism are genetically different in these three organs in A. thaliana. In addition, these mutants showed normal phototropic responses, suggesting that SGR4, SGR5 and SGR6 genes are specifically involved in gravity perception and/or gravity signal transduction for the shoot gravitropic response. PMID- 9210332 TI - Nucleotide sequence and transcriptional analysis of the flanking region of the gene (spb) for the trans-acting factor that controls light-mediated expression of the puf operon in Rhodobacter sphaeroides. AB - We recently reported the existence of a trans-acting factor (SPB) in Rhodobacter sphaeroides that repressed the expression of the puf operon during illumination. SPB was somewhat homologous to HvrA of Rhodobacter capsulatus, but these proteins appear to have functionally different properties. We now report an analysis of the flanking region of spb in the genome of R.sphaeroides, and we show that spb is a positional counterpart of hvrA of R. capsulatus. The region directly downstream of spb was found to contain three genes, two of which were highly homologous to orf5 and ahcY in R. capsulatus. However, a gene corresponding to hvrB, which controls the expression of orf5 and ahcY in R. capsulatus, was absent in R. sphaeroides. The level of the transcript of ahcY did not change in cells grown under photosynthetic and by respiratory conditions. By contrast, orf5 was transcribed at a higher rate in photosynthetically grown cells under high intensity light than under low-intensity light, indicating features of transcription different from those in R. capsulatus. A third gene, orf318, which was absent in the corresponding region of R. capsulatus, encoded an amino acid sequence that was significantly homologous to the consensus sequence of RfaI and RfaJ of E.coli, which are glycosyl transferases involved in the synthesis of lipopolysaccharide. orf318 was transcribed in the opposite direction to ahcY, and at only a low level, under all conditions tested. PMID- 9210333 TI - Herbicide safener-inducible gene expression in Arabidopsis thaliana. AB - The potential use of a new chemical-inducible gene expression system in Arabidopsis thaliana has been examined. The system is based on the maize In2-2 promoter which is activated by benzenesulfonamide herbicide safeners. Plants transformed with the beta-glucuronidase (gus) reporter gene under the control of the In2-2 promoter were grown in the presence of different safeners and the induced GUS activity pattern was studied histochemically. In the absence of safeners, the In2-2 promoter was not active. Application of different safeners induced distinct gus expression patterns, including expression in the root, hydathodes, and the shoot apical meristem. Plants maintained continuously on inducing concentrations of the safeners were retarded in growth. The growth inhibition effects of the Sa5 safener could be overcome in a sulfonylurea resistant background. In2-2 promoter activity could also be induced by the sulfonylurea herbicide chlorsulfuron. In the sulfonylurea-resistant background, which derives from herbicide-resistant acetolactate synthase activity, induction of the In2-2 promoter by chlorsulfuron was lower. Furthermore, branched-chain amino acids, known to inhibit acetolactate synthase activity, also induced In2-2 promoter activity. Our data suggest a strong correlation between In2-2 expression and inhibition of the acetolactate synthase activity. PMID- 9210335 TI - Cloning of the pumpkin ascorbate oxidase gene and analysis of a cis-acting region involved in induction by auxin. AB - A genomic clone encoding ascorbate oxidase was isolated from pumpkin (Cucurbita sp.). This gene is consisted of four exons and three introns. Analyses of the promoter fusion to beta-glucuronidase reporter gene by transient expression assay in pumpkin fruit tissues suggested the existence of a cis-acting region responsible for auxin regulation. PMID- 9210337 TI - Continuous sucrose hydrolysis by yeast cells immobilized to wool. AB - A novel immobilized biocatalyst with invertase activity was prepared by adhesion of yeast cells to wool using-glutaraldehyde. Yeast cells could be immobilized onto wool by treating either the yeast cells or wool or both with glutaraldehyde. Immobilized cells were not desorbed by washing with 1 M KCl or 0.1 M buffers. pH 3.5-7.5. The biocatalyst shows a maximum enzyme activity when immobilized at pH 4.2-4.6 and 7.5-8.0. The immobilized biocatalyst was tested in a tubular fixed bed reactor to investigate its possible application for continuous full-scale sucrose hydrolysis. The influence of temperature, sugar concentration and flow rate on the productivity of the reactor and on the specific productivity of the biocatalyst was studied. The system demonstrates a very good productivity at a temperature of 70 degrees C and a sugar concentration of 2.0 M. The increase of the volume of the biocatalyst layer exponentially increases the productivity. The productivity of the immobilized biocatalyst decreases no more than 50% during 60 days of continuous work at 70 degrees C and 2.0 M sucrose, but during the first 30 days it remains constant. The cumulative biocatalyst productivity for 60 days was 4.8 x 10(3) kg inverted sucrose/kg biocatalyst. The biocatalyst was proved to be fully capable of continuous sucrose hydrolysis in fixed-bed reactors. PMID- 9210336 TI - Isolation and characterization of cDNA clones with enhanced expression in tobacco plants expressing the rice homeobox gene, OSH1. AB - We have used subtractive hybridization to isolate cDNA clones whose expression were up-regulated in transgenic tobacco ectopically expressing the rice homeobox gene, OSH1. Thirty-nine distinct cDNA clones, which we term HRGs (Homeobox Regulated Genes), were identified. Some of them were specifically expressed in transformants, indicating that their expression was possibly regulated by transgene. PMID- 9210338 TI - Cell-dislodging methods under serum-free conditions. AB - In this work, a BHK21 clone producing a fusion protein consisting of a recombinant human IgG molecule with a cytokine tail, growing in a protein-free medium, was used to test several alternatives to avoid the use of serum for trypsin inactivation, currently used in cell dislodging. These included (1) trypsin inactivated with soybean trypsin inhibitor (STI); (2) cell dissociation solution instead of trypsin; (3) dispase instead of trypsin; (4) trypsin inactivated with fetal calf serum (positive control); (5) non-inactivated trypsin (negative control). Use of a centrifugation step was also tested for each alternative. Results indicate that the best method regarding cell growth, viability and adherent fraction is to use trypsin inactivated with STI followed by a centrifugation step. For all methods tested, the utilization of a centrifugation step always led to improved results. The optimal proportion for total trypsin inactivation is 1:1 trypsin (0.2% w/v) to STI (1 mg ml-1). equivalent to 2 mg trypsin to 1 mg STI. No toxic effect was observed for STI at the concentrations used. Long-term subculturing with this new, alternative dislodging method did not affect cell growth, viability and productivity. PMID- 9210339 TI - Structural and biochemical properties of glycosylated and deglycosylated glucose oxidase from Penicillium amagasakiense. AB - Glucose oxidase from Penicillium amagasakiense was purified to homogeneity by ion exchange chromatography and deglycosylated with endoglycosidase H. On the basis of gas chromatography and sodium dodecyl sulphate/polyacrylamide gel electrophoretic (SDS-PAGE) analyses, the protein-bound high-mannose-type carbohydrate moiety corresponded to 13% of the molecular mass of glycosylated glucose oxidase. A total of six N-glycosylation sites per dimer were determined from the N-acetylglucosamine content. The enzymatically deglycosylated enzyme contained less than 5% of the original carbohydrate moiety. A molecular mass of 130 kDa (gel filtration) and 133 kDa (native PAGE) was determined for the dimer and 67 kDa (SDS-PAGE) for the monomer of the deglycosylated enzyme. The N terminal sequence, which has not-been published for glucose oxidase from P. amagasakiense to date and which showed less than 50% homology to the N terminus of glucose oxidase from Aspergillus niger, and the amino acid composition were not altered by the deglycosylation. Deglycosylation also did not affect the kinetics of glucose oxidation or the pH and temperature optima. It also did not increase the susceptibility of the enzyme to proteolytic degradation. However, deglycosylated glucose oxidase exhibited decreased pH and thermal stability. The thermal stability of both enzymes was shown to be dependent on the buffer concentration and was enhanced by certain additives, particularly 1 M (NH4)2SO4, which stabilised glucose oxidase 100- to 300-fold at 50 degrees C and pH 7-8, and 2 M KF, which stabilised the enzyme up to 36-fold at 60 degrees C and pH 6. In sodium acetate buffer, changes in pH (4-6) affected the affinity for glucose but had no effect on the Vmax of the reaction. In contrast, in TRIS buffer, pH 8, a 10-fold decrease in Vmax and a 2-fold decrease in K(m) were observed. PMID- 9210340 TI - Pyranose 2-oxidase from Phanerochaete chrysosporium--further biochemical characterisation. AB - Pyranose 2-oxidase (P2O) was purified 43-fold to apparent homogeneity from the basidiomycete Phanerochaete chrysosporium using liquid chromatography on phenyl Sepharose, Mono Q (twice) and phenyl Superose. The native enzyme has a molecular mass of about 250 kDa (based on native PAGE) and is composed of four identical subunits of 65 kDa. It contains three isoforms of isoelectric point (pI) 5.0, 5.05 and 5.15 and does not appear to be a glycoprotein. P2O is optimally stable at pH 8.0 and up to 60 degrees C. It is active over a broad pH range (5.0-9.0) with maximum activity at pH 8.0-8.5 and at 55 degrees C, and a broad substrate specificity. D-Glucose is the preferred substrate, but 1-beta-aurothioglucose, 6 deoxy-D-glucose, L-sorbose, D-xylose, 5-thioglucose, D-glucono-1,5-lactone, maltose and 2-deoxy-D-glucose are also oxidised at relatively high rates. A Ping Pong Bi Bi mechanism was demonstrated for the P2O reaction at pH 8.0, with a catalytic constant (kcat) of 111.0 s-1 and an affinity constant (Km) of 1.43 mM for D-glucose and 83.2 microM for oxygen. Whereas the steady-state kinetics for glucose oxidation were unaffected by the medium at pH > or = 7.0, at low pH both pH and buffer composition affected the P2O kinetics with the kcat/K(m) value decreasing with decreasing pH. The greatest effect was observed in acetate buffer (0.1 M, pH 4.5), where the kcat decreased to 60.9 s-1 and the K(m) increased to 240 mM. The activity of P2O was completely inhibited by 10 mM HgCl2, AgNO3 and ZnCl2, and 50% by lead acetate, CuCl2 and MnCl2. PMID- 9210341 TI - Purification of alpha-amylase by specific elution from anti-peptide antibodies. AB - Chimeric alpha-amylase, produced by recombinant yeast cells, was purified by immunoaffinity chromatography by use of an anti-peptide antibody and an eluent containing an antigen peptide. Chimeric alpha-amylase was adsorbed by the antibody against the peptide corresponding to the C-terminal region of target alpha-amylase, and specifically eluted by the eluent containing the antigen peptide used for immunization. A low concentration of the peptide could competitively elute adsorbed alpha-amylase, and the rate-limiting step of the elution was mass transfer of desorbed alpha-amylase. With this specific method, target proteins can be effectively eluted, and highly purified under mild conditions, from the antibody ligand showing a high-affinity for the adsorption step PMID- 9210342 TI - Controlled secretion into the culture medium of a hybrid beta-glucanase by Acetobacter methanolicus mediated by the kil gene of Escherichia coli located on a Tn5-derived transposon. AB - A Tn5-based transposon bearing the kil gene (killing protein), mediating controlled export of periplasmic proteins into the culture medium, was constructed (Tn5-KIL3). This transposon contained the kil gene of the ColEl plasmid under the growth-phase-dependent promoter of the fic gene (filamentation induced by cAMP) of Escherichia coli, an interposon located upstream of kil, a kanamycin/neomycin-resistance gene, a multiple cloning site and the mob site. The transposition of Tn5-KIL3 to Acetobacter methanolicus showed a moderate transposition frequency (10(-5) -10(-6). By insertion of a Bacillus hybrid beta glucanase (bgl) as a model protein into the transposon (Tn5-LF3) it was shown that the secretion function as well as the gene of the target protein had been transferred to and stably integrated into the chromosome of A. methanolicus, and that the transposition of Tn5-LF3 was non-specific. beta-Glucanase was highly overexpressed and secreted into the medium during stationary phase. Total and extra-cellular production of beta-glucanase varied depending on the integration site of the transposon. The viability of the bacterial cells was not affected, and cell lysis did not occur. PMID- 9210343 TI - Protein A as a fusion partner for the expression of heterologous proteins in Lactobacillus. AB - An expression system based on the Staphylococcus aureus protein A gene (spa) was developed to allow the production and export of proteins in Lactobacillus. Plasmid shuttle vectors were constructed that carried the eZZ gene, a synthetic gene based on the Protein A gene (spa) but lacking the carboxy-terminal membrane anchoring region. A gene fusion was created between the eZZ gene and the VD4 region of a chlamydial major outer-membrane protein gene. Expression studies demonstrated the recognition of the spa regulatory signals by several Lactobacillus, with the recombinant protein being expressed (from 0.1 microgram of EZZVD4 fusion protein per ml in L. plantarum up to 10 micrograms of EZZ protein per ml in L. fermentum) and exported (levels up to 20% in L. fermentum) in several Lactobacillus strains. PMID- 9210344 TI - Plasmids for efficient single-copy gene cloning into gdh2 and trpC of Bacillus megaterium DSM319 and QM B1551. AB - We constructed integrative plasmids to place xylA-lacZ indicator gene fusions into two different loci of the Bacillus megaterium chromosome, gdh2 and trpC, in lac mutants of strains DSM 319 and QM B1551, which differ markedly. Single crossover integration was achieved in all cases while double crossovers occurred only in gdh2 of DSM 319 and QM B1551 and in trpC of QM B1551. Neither of the loci affected regulation of the xylA-lacZ fusions. These results confirm the suitability of the two gene loci for single-copy cloning. PMID- 9210345 TI - Influence of aeration and carbon source on production of microcin B17 by Escherichia coli ZK650. AB - Previous studies [Connell et al. (1987) Mol Microbiol 1: 195-201] have shown that expression of the microcin B17 (MccB17) promoter is inversely related to the growth rate of the culture, when slower growth was brought about by limitation of sources of carbon, nitrogen or phosphorus. When we used oxygen limitation to decrease growth in a glucose-based chemically defined medium, we found specific MccB17 production to be positively related to growth rate and extent. On the other hand, when we examined various nutritional variations of media, specific production of MccB17 showed a negative relationship to growth rate and extent, as would be predicted by the findings of Connell et al. (1987). Glucose, glycerol and acetate were found to repress MccB17 production; succinate was not repressive. Succinate is an excellent carbon source for production of MccB17 since high levels can be used with no or little interference in product synthesis. PMID- 9210346 TI - 3-Ketoglycoside-mediated metabolism of sucrose in E. coli as conferred by genes from Agrobacterium tumefaciens. AB - Escherichia coli strains that did not have the ability to use sucrose as a sole carbon source gained this ability after receiving a cloned fragment of DNA from Agrobacterium tumefaciens. No invertase was detected in the sucrose-metabolizing E. coli, but evidence for the activity of certain enzymes, known to be produced by biotype 1 strains of Agrobacterium, were found. Evidence was found for the presence of D-glucoside 3-dehydrogenase (G3DH) and alpha-3-ketoglucosidase. The activity of enzyme extracts on 3-ketosucrose also indicated that 3-ketoglucose reductase, or some enzyme that acts on 3-ketoglucose, was present in the Suc+ E. coli as well. The fragment was found to complement a G3DH mutant of A. tumefaciens and was also found to confer chemotaxis towards sucrose in E. coli. PMID- 9210347 TI - Cell-linked and extracellular cholesterol oxidase activities from Rhodococcus erythropolis. Isolation and physiological characterization. AB - Rhodococcus erythropolis cells growing in a cholesterol-free glycerol-containing mineral medium displayed very low levels of a cell-wall-bound cholesterol oxidase activity. Addition of cholesterol induced a marked increase in the synthesis of this enzyme, which reached a maximum within 6 days and was subsequently followed by the appearance of extracellular cholesterol oxidase in the culture broth. Significant levels of induction were only achieved when cholesterol emulsified with Tween 80. The presence of chloramphenicol at the time of induction completely prevented the emergence of both enzymatic forms, suggesting the requirement of de novo protein synthesis. Upon transfer of cholesterol-growing cultures to fresh medium lacking cholesterol, the extracellular cholesterol oxidase was quickly erased, while the activity of the particulate enzyme decreased sharply. The electrophoretic pattern on native Western blotting as well as on sodium dodecyl sulphate/polyacrylamide gels, together with kinetic data, strongly support the idea that the particulate and extracellular cholesterol oxidases are two different forms of the same enzyme with an estimated molecular mass of 55 kDa. PMID- 9210348 TI - Expanded bed adsorption for recovery of renatured human recombinant interleukin 8 from Escherichia coli inclusion bodies. AB - Expanded bed adsorption is a chromatography technique based on stable fluidization, designed for large scale recovery of proteins directly from particulate-containing feedstocks. Described in this article is the use of expanded bed adsorption for recovery of renatured recombinant human interleukin 8 from Escherichia coli inclusion bodies, using a cation exchange adsorbent. The yield of interleukin 8 from the expanded bed was approximately 100% and the purification factor was approximately 4. Following each interleukin 8 purification a cleaning in place procedure was performed with a commercial cleaner. The results show that the adsorbent can be used at least 50 times without any significant loss of function. PMID- 9210350 TI - Partitioning and purification of monoclonal antibodies in aqueous two-phase systems. AB - The partition behaviour of pure IgG in aqueous two-phase systems was examined in order to investigate the effects of changes in phase properties on the partition coefficient, K. Factors such as molecular weight (M.W.) of PEG, pH, and concentration of NaCl and other salts were all found to influence K. Due to the high level of interaction between the phase components, optimal conditions were found using a factorial design. Using this methodology it was possible to find conditions which gave extremely high values of K for the IgGs (> 100). Partition behaviour of some potential contaminants, BSA and transferrin, were also studied. All the components were characterized for hydrophobicity, pI and M.W. Conditions were chosen under which the partition coefficients of the contaminants was low (pH 6, 15% PEG 1450, 14% Phosphate and 12% NaCl). Precipitation of IgG was also minimized. These conditions were chosen for the extraction of IgG from the contaminants in a crude concentrated supernatant. The presence of culture media contaminants had a strong effect in the systems and hence on the partition of IgG which was lower than for pure IgG. The ratio of KIgG/Kcont was > 25. The conditions chosen for the first forward extraction into the PEG rich phase were 15% PEG 1450, 14% phosphate 12% NaCl and pH 5.5. Virtually all IgG partitioned to the top phase and the contaminants to the bottom phase. Optimal conditions for the back extraction of the IgGs into a bottom salt phase were also found (addition of 14% phosphate with no NaCl). An industrial serum free, crude, concentrated culture supernatant of hybridoma produced murine IgG, with a relatively low level of protein contaminants (14% IgG purity) was processed in this system. After the back extraction the contaminants were reduced 18 fold giving IgG with 80% purity. A 5.9 fold purification was obtained out of a 7.3 maximum possible (at 100% pure IgG). All of the IgG was recovered. PMID- 9210349 TI - Evaluation of affinity and pseudo-affinity adsorption processes for penicillin acylase purification. AB - Affinity ligand (6-Aminopenicillanic acid, Amoxycillin, Ampicillin, Benzylpenicillin and 4-Phenylbutylanzine) of penicillin acylase (EC 3.5.1.11) were attached to hydrophilic gels like Sepharose 4B-CNBr and Minileak 'medium'. Ampicillin and 4-Phenylbutylamine were the affinity ligands that presented the higher concentrations attached to both gels. Penicillin acylase adsorption on these affinity gels was mainly dependent on the activated group of the gel, the affinity ligand attached and the experimental conditions of enzyme adsorption. Under affinity conditions only the ligands Amoxycillin, Ampicillin and 4 Phenylbutylamine, immobilized on Minileak, adsorbed the enzyme from osmotic shock extracts at different pH values. These affinity ligand systems were characterized by low adsorption capacities of penicillin acylase activity (1.2-2.1 IU mL-1 gel) and specific activity (1.5-2.9 IU mg-1 prot). Under pseudo-affinity conditions all the ligands attached both activated to gels (Sepharose 4B-CNBr and Minileak) adsorbed the enzyme. The affinity gels were characterized by higher values of adsorption capacity (3.7 and 55.6 IU mL-1 gel) and adsorbed specific activity (2.0 and 6.1 IU mg-1 prot) than those observed under affinity conditions. The space arm of Minileak gel, shown to be fundamental to enzyme adsorption under affinity conditions, preferentially adsorbed proteins in relation to the enzyme under pseudo-affinity conditions. However, this effect was partially minimized when the gel was derivatized by the affinity ligands at concentrations higher than 6 mumol mL-1 gel. Ampicillin was the affinity ligand that presented the best results for specific adsorption of penicillin acylase under affinity and pseudo affinity adsorption processes. The Sepharose 4B-CNBr derivatized gel also presented a good adsorption capacity of enzyme activity (26.8 IU mL-1 gel) under pseudo-affinity adsorption processes. PMID- 9210352 TI - Prognostic factors in colorectal carcinomas. AB - Colorectal carcinoma remains a leading cause of cancer morbidity and mortality. Various clinical signs and pathologic factors have been shown to have a bearing on a patient's prognosis. Some of these factors, such as extent of disease (stage) and histologic grade, are generally accepted, while others, primarily biologic and molecular markers, have been proposed recently and remain controversial. The authors describe both more established and newly proposed variables, reviewing multivariate analyses to examine their relative importance. The recommendations of the Association of Directors of Anatomic and Surgical Pathology for the reporting of colorectal carcinomas are presented. PMID- 9210351 TI - Pathologic prognostic factors for patients with breast carcinoma. Which factors are important. AB - Current technologies can identify subsets of patients who are at greater risk for developing recurrent carcinoma. This article presents conventional and generally accepted pathologic features of breast carcinoma that allow breast carcinoma patients to be placed into low-risk or high-risk categories for recurrence-free or overall survival. Also, the role of flow cytometry, estrogen-progesterone receptor measurement, tumor angiogenesis, and selection oncoprotein expression, such as c-erbB2, are reviewed. PMID- 9210353 TI - Prognostic indicators for cancer. Gastric cancer. AB - The recent shift in the population from patients presenting with gastric cancer to patients presenting with early-stage lesions has led to renewed interest in identifying prognostic factors for this type of tumor. Conveniently for surgeons, prognostic factors can be divided into groups that are assessed preoperatively, intraoperatively, and postoperatively. Despite the explosion of interest in genetic and molecular markers for gastric cancer, the feature best correlated with patient survival continues to be tumor stage at the time of diagnosis. PMID- 9210354 TI - Prognostic factors in esophageal cancer. AB - Clinical and pathologic factors relevant to the prognosis of esophageal cancer are reviewed in this article. Clinical factors discussed include endoscopic, radiologic, and surgical findings. The importance of Barrett's esophagus and the role of endoscopic screening in the diagnosis of dysplasia and the prevention of adenocarcinoma are evaluated. Pathologic factors include the traditional ones of tumor type, stage, and grade as well as newer tumor markers related to DNA content, rate of cell proliferation, oncogenes, and tumor suppressor genes. PMID- 9210355 TI - Prognostic factors in pancreatic carcinoma. AB - Pancreatic cancer is a relatively common malignancy. Its gravity is underscored by the low overall cure rates. A number of clinical, pathologic, and molecular factors have been identified that predict survival of patients with this neoplasm. These factors are reviewed and analyzed. PMID- 9210356 TI - Prognostic factors in thyroid carcinoma. AB - Thyroid carcinomas range from the most indolent to the most aggressive human malignancies. A variety of clinical and pathologic factors can be used to help predict behavior and determine appropriate therapy. New molecular biologic techniques show promise in further refining the assessment of prognosis in thyroid cancer patients. PMID- 9210357 TI - Prognostic factors in malignant melanoma. AB - The clinical spectrum and biologic behavior of melanoma are heterogeneous. Several clinical factors may include sex, location, and age. Histologic prognostic factors may include tumor type, tumor thickness, and mitotic index in relationship to the primary tumor. With respect to metastatic melanoma, the prognostic factors may include lymph node status, number of lymph nodes, and extracapsular extension. Cytogenetic and molecular determinants of progression of melanoma may aid the current prognostic factors when they are established more firmly. Current surgical treatment is determined by the clinical and histologic factors. PMID- 9210358 TI - Prognostic factors for cutaneous squamous cell and basal cell carcinoma. Determinants of risk of recurrence, metastasis, and development of subsequent skin cancers. AB - Squamous cell carcinoma and basal cell carcinoma are the most common cancers in humans. This article discusses general prognostic factors for these nonmelanocytic skin cancers. Anatomic and histologic considerations for both carcinomas are also presented. PMID- 9210359 TI - Prognostic factors and management of carcinomas of the gallbladder and extrahepatic bile ducts. AB - Cancers of the biliary tract are uncommon but aggressive malignancies that pose difficult problems in diagnosis and management. Long-term survival with these cancers is limited by their propensity for local invasion, so that pathologic stage becomes a major prognostic factor, and by their ability to cause biliary obstruction and sepsis and interfere with hepatic function. In selected patients, surgical resection offers the possibility of cure, but effective palliation is often the principal goal of treatment. Radiologic and endoscopic modalities thus often play a major role in patient management. PMID- 9210360 TI - Plant evolution: the dominance of maize. AB - The gene teosinte branched 1 controls major differences in architecture between cultivated maize and its wild ancestor. The differences correlate with different amounts of gene product. PMID- 9210361 TI - Ageing: levelling of the grim reaper. AB - Recent observations of a levelling of the death rate in extreme old age, in both experimental species and humans, are posing difficult problems for evolutionary biologists, in particular about the evolution of the post-reproductive period. PMID- 9210362 TI - Tropical rainforests: diversity begets diversity. AB - Tropical rainforests exhibit an extraordinarily high level of biological diversity. A new study shows that the patterns of seedling survival surrounding parent trees are responsible in large part for this amazing diversity. PMID- 9210363 TI - Axon-glia interactions: building a smart nerve fiber. AB - Myelinated nerve fibers are designed in an optimal manner which requires tuning of conduction time with millisecond precision. This involves the highly coordinated differentiation of axons and myelin-forming glial cells; the nature of the signals involved in this axon-glial cell dance are beginning to be elucidated. PMID- 9210364 TI - Actin cytoskeleton: are FH proteins local organizers? AB - The FH proteins, defined by the presence of 'formin homology' regions, are important for a number of actin-dependent processes, including polarized cell growth and cytokinesis. They are large, probably multi-domain, proteins and their function may be in part mediated by an interaction with profilin. PMID- 9210365 TI - Retinal physiology: adapting to the changing scene. AB - The retina needs to process visual information under a wide range of conditions, a feat facilitated by gain controls. Recent results have provided new insights into one such gain control, which enables the retina to adapt to wide variations in the level of contrast in the visual scene. PMID- 9210366 TI - Retinal development: communication helps you see the light. AB - Recent studies suggest that interactions between neurons, glial cells and endothelial cells are critical in determining the structure of the retina and the optic nerve. Dysregulation of these interactions can lead to disruption of retinal architecture and impairment of vision. PMID- 9210367 TI - Binocular rivalry: ambiguities resolved. AB - For over 100 years, binocular rivalry was seen as the result of competition between the two eyes, involving reciprocal suppression of retinal inputs. Now it emerges that rivalry reflects alternating perceptual interpretations that are represented in the firing patterns of cells in the temporal visual cortex. PMID- 9210368 TI - Carcinogenesis: a balance between beta-catenin and APC. AB - A protein first identified by its association with cadherin cell adhesion molecules, beta-catenin, has been implicated in carcinogenesis. In a number of different types of cancer, signalling through beta-catenin is upregulated either by direct mutation of beta-catenin or loss of negative regulation by the APC tumor suppressor protein. PMID- 9210369 TI - RNA splicing: out of the loop. AB - Recent functional analysis of catalytic and exon-binding domains from group II autocatalytic introns has revealed haunting similarities with small nuclear RNA sequences in the spliceosome. PMID- 9210370 TI - RNA editing: rewriting receptors. AB - The type of RNA editing that converts adenosine to inosine in double-stranded RNA generates different isoforms of subunits of the ionotropic glutamate-gated ion channel receptors. Recently, it has been reported that the pre-mRNA of the serotonin 2C receptor can be edited by the same mechanism. PMID- 9210371 TI - Plant meristems: cell signalling keeps the balance. AB - Genetic and molecular analysis in Arabidopsis has identified components of a putative cell signalling pathway that appears to regulate the balance between stem cell proliferation and fate specification in meristems. PMID- 9210372 TI - Cytokines: shared receptors, distinct functions. AB - That the signal transduction pathways used by the cytokines IL-2 and IL-15 are identical would suggest that these cytokines have redundant roles in lymphoid development; instead, IL-2 is the guardian of thymus-derived T-cell homeostasis, while interleukin-15 promotes extrathymic development of T and NK cells. PMID- 9210373 TI - Abrupt learning and retinal size specificity in illusory-contour perception. AB - BACKGROUND: In behavioral studies of learning, a distinction is commonly made between gradual and abrupt improvements in performance. The learning of perceptual and motor skills is often characterized by gradual, incremental improvement, and is found not to generalize over stimulus manipulations such as changes in the size or location of the retinal image. In contrast, marked improvement in performance can occur suddenly - a phenomenon which has been termed 'insight'. Consequently, the brain mechanisms subserving the two types of learning are commonly thought of as distinct. Here, we examine learning of a perceptual task in which improvement appears to exhibit characteristics of both gradual and abrupt learning. RESULTS: We describe experiments on illusory-contour perception in which the observers underwent an abrupt, dramatic improvement in performance, resembling an incident of insight. At the same time, however, the phenomenon showed a degree of stimulus-specificity that was previously thought to characterize incremental, gradual learning. The improvement was triggered only by specific visual stimuli, whereas other, quite similar, stimuli were found to be ineffective for training; the learning did not generalize to a new retinal image size, and re-training was necessary for different-sized images. CONCLUSIONS: The juxtaposition of abrupt and stimulus-specific learning that we observed suggests that the distinction between the two forms of learning needs to be revised. Rather than postulating two distinct mechanisms, incremental and insightful learning need to be addressed within a single framework. In particular, the findings suggest that learning may involve interactions between multiple levels of representations of the stimulus. PMID- 9210374 TI - Caenorhabditis elegans CED-4 stimulates CED-3 processing and CED-3-induced apoptosis. AB - BACKGROUND: Programmed cell death or apoptosis is a key feature of normal development, tissue homeostasis and disease progression in metazoans. Genetic studies in the nematode C. elegans have identified three key genes involved in apoptosis, ced-3, ced-4 and ced-9. Expression of ced-3 and ced-4 is required for the induction of cell death, whereas expression of ced-9 is necessary to inhibit cell death. The precise mechanism by which these genes influence the life or death decision of a cell is not known. In this study, we have expressed the genes in an insect cell line to explore their role in the apoptotic pathway. RESULTS: Co-expression of ced-4 with ced-3 in insect cells stimulated both the induction and the level of CED-3-mediated apoptosis. Stimulation of CED-3-dependent apoptosis by CED-4 was accompanied by accelerated processing of CED-3, which was dependent on the presence of a wild-type CED-3 prodomain and a conserved lysine residue within a putative ATP/GTP-binding motif of CED-4. Co-expression of ced-9 with ced-4 and ced-3 inhibited the ability of CED-4 to stimulate CED-3 processing and CED-3-dependent apoptosis. Although a temperature-sensitive CED-9 mutant was unable to block CED-4 activity and failed to associate with CED-4, a deletion mutant of CED-9 lacking the carboxy-terminal hydrophobic domain could associate with CED-4 and block CED-4 activity. CONCLUSIONS: Our results establish a role for CED-4 in the processing of CED-3 and the stimulation of CED-3-induced apoptosis. Furthermore, we show that CED-9 achieves its anti-apoptotic effect by associating with CED-4 and blocking the ability of CED-4 to process CED-3. PMID- 9210375 TI - A Cdc42 target protein with homology to the non-kinase domain of FER has a potential role in regulating the actin cytoskeleton. AB - BACKGROUND: Members of the Rho family of small GTPases have been shown to have a diverse role in cell signalling events. They were originally identified as proteins that, by regulating the assembly of the actin cytoskeleton, are important determinants of cell morphology, and have recently been shown to be involved in transcriptional activation by the JNK/SAPK signalling pathway. In order to understand the mechanisms underlying the effects of Rho GTPases on these processes, the yeast two-hybrid system has been used to identify proteins that bind to an activated mutant of Cdc42, a Rho-family member. RESULTS: A cDNA encoding a previously unidentified Cdc42 target protein, CIP4, which is 545 amino acids long and contains an SH3 domain at its carboxyl terminus, was cloned from a human B-cell library. The amino terminus of CIP4 bears resemblance to the non kinase domain of the FER and Fes/Fps family of tyrosine kinases. In addition, similarities to a number of proteins with roles in regulating the actin cytoskeleton were noticed. CIP4 binds to activated Cdc42 in vitro and in vivo and overexpression of CIP4 in Swiss 3T3 fibroblasts reduces the amount of stress fibres in these cells. Moreover, coexpression of activated Cdc42 and CIP4 leads to clustering of CIP4 to a large number of foci at the dorsal side of the cells. CONCLUSIONS: CIP4 is a downstream target of activated GTP-bound Cdc42, and is similar in sequence to proteins involved in signalling and cytoskeletal control. Together, these findings suggest that CIP4 may act as a link between Cdc42 signalling and regulation of the actin cytoskeleton. PMID- 9210376 TI - The complex containing actin-related proteins Arp2 and Arp3 is required for the motility and integrity of yeast actin patches. AB - BACKGROUND: Structural modeling and biochemical experiments in vitro have implicated a multi-protein complex containing two actin-related proteins, Arp2 and Arp3, as a potential actin-filament nucleation factor. This 'Arp2/3 complex' has been identified in Acanthamoeba and human cells and has been shown to localize to regions involved in actin-based motility, such as the leading edge of moving cells and the 'tail' of actin that forms behind the intracellular pathogen Listeria. The function of this complex in vivo has not been characterized, however, and the sequences of the non-actin-related subunits remain to be determined. RESULTS: An Arp3 homologue from the budding yeast Saccharomyces cerevisiae was found to localize to cortical actin patches, highly motile structures that concentrate at sites of polarized growth during the yeast cell cycle. A conditional arp3 mutant allele inhibited cortical actin motility at the restrictive temperature and eventually disrupted actin patches. Most Arp3 protein is found in a multi-protein complex; we purified this complex and determined the sequences of each of the protein subunits using a high-accuracy mass peptide mapping technique. The proteins found in the complex are similar to those in the Acanthamoeba and human Arp2/3 complexes except that the yeast complex lacks a 40 kDa subunit, which is therefore not required for the structural integrity of the complex. CONCLUSIONS: The Arp2/3 protein complex is conserved from yeast to man, and in yeast the complex is required in vivo for the motility and integrity of cortical actin patches. We hypothesize that these patches may move by a Listeria like mechanism driven by actin polymerization. PMID- 9210377 TI - The mago nashi gene is required for the polarisation of the oocyte and the formation of perpendicular axes in Drosophila. AB - BACKGROUND: Drosophila axis formation requires a series of inductive interactions between the oocyte and the somatic follicle cells. Early in oogenesis, Gurken protein, a member of the transforming growth factor alpha family, is produced by the oocyte to induce the adiacent follicle cells to adopt a posterior cell fate. These cells subsequently send an unidentified signal back to the oocyte to induce the formation of a polarised microtubule array that defines the anterior posterior axis. The polarised microtubules also direct the movement of the nucleus and gurken mRNA from the posterior to the anterior of the oocyte, where Gurken signals a second time to induce the dorsal follicle cells, thereby polarising the dorsal-ventral axis. RESULTS: In addition to its previously described role in the localisation of oskar mRNA, the mago nashi gene is required in the germ line for the transduction of the polarising signal from the posterior follicle cells. Using a new in vivo marker for microtubules, we show that mago nashi mutant oocytes develop a symmetric microtubule cytoskeleton that leads to the transient localisation of bicoid mRNA to both poles. Furthermore, the oocyte nucleus often fails to migrate to the anterior, causing the second Gurken signal to be sent in the same direction as the first. This results in a novel phenotype in which the anterior of the egg is ventralised and the posterior dorsalised, demonstrating that the migration of the oocyte nucleus determines the relative orientation of the two principal axes of Drosophila. The mago nashi gene is highly conserved from plants to animals, and encodes a protein that is predominantly localised to nuclei. CONCLUSIONS: The mago nashi gene plays two essential roles in Drosophila axis formation: it is required downstream of the signal from the posterior follicle cells for the polarisation of the oocyte microtubule cytoskeleton, and has a second, independent role in the localisation of oskar mRNA to the posterior of the oocyte. PMID- 9210378 TI - Cooperative activation of IP3 receptors by sequential binding of IP3 and Ca2+ safeguards against spontaneous activity. AB - BACKGROUND: Ca2+ waves allow effective delivery of intracellular Ca2+ signals to cytosolic targets. Propagation of these regenerative Ca2+ signals probably results from the activation of intracellular Ca2+ channels by the increase in cytosolic [Ca2+] that follows the opening of these channels. Such positive feedback is potentially explosive. Mechanisms that limit the spontaneous opening of intracellular Ca2+ channels are therefore likely to have evolved in parallel with the mechanism of Ca2+-induced Ca2+ release. RESULTS: Maximal rates of 45Ca2+ efflux from permeabilised hepatocytes superfused with medium in which the [Ca2+] was clamped were cooperatively stimulated by inositol 1,4,5-trisphosphate (IP3). A minimal interval of approximately 400 msec between IP3 addition and the peak rate of Ca2+ mobilisation indicate that channel opening does not immediately follow binding of IP3. Although the absolute latency of Ca2+ release was unaffected by further increasing the IP3 concentration, it was reduced by increased [Ca2+]. CONCLUSIONS: We propose that the closed conformation of the IP3 receptor is very stable and therefore minimally susceptible to spontaneous activation; at least three (probably four) IP3 molecules may be required to provide enough binding energy to drive the receptor into a stable open conformation. We suggest that a further defence from noise is provided by an extreme form of coincidence detection. Binding of IP3 to each of its four receptor subunits unmasks a site to which Ca2+ must bind before the channel can open. As IP3 binding may also initiate receptor inactivation, there may be only a narrow temporal window during which each receptor subunit must bind both of its agonists if the channel is to open rather than inactivate. PMID- 9210379 TI - Transposon-generated 'knock-out' and 'knock-in' gene-targeting constructs for use in mice. AB - The conventional technique for targeted mutation of mouse genes entails placing a genomic DNA fragment containing the gene of interest into a vector for fine mapping, followed by cloning of two genomic arms around a selectable neomycin resistance cassette in a vector containing thymidine kinase [1]; this generally requires 1-2 months of work for each construct. The single 'knock-out' construct is then transfected into mouse embryonic stem (ES) cells, which are subsequently subjected to positive selection (using G418 to select for neomycin-resistance) and negative selection (using FIAU to exclude cells lacking thymidine kinase), allowing the selection of cells which have undergone homologous recombination with the knockout vector. This approach leads to inactivation of the gene of interest [2]. Recently, an in vitro reaction was developed, on the basis of the yeast Ty transposon, as a useful technique in shotgun sequencing [3]. An artificial transposable element, integrase enzyme and the target plasmid are incubated together to engender transposition. The DNA is then purified, and subsequently electroporated into bacteria. The transposon and the target plasmid bear distinct antibiotic resistance markers (trimethoprim and ampicillin, respectively), allowing double selection for transposition events. In the present study, we have modified this system to allow the rapid, simultaneous generation of a palette of potential gene targeting constructs. Our approach led from genomic clone to completed construct ready for transfection in a matter of days. The results presented here indicate that this technique should also be applicable to the generation of gene fusion constructs [4-8], simplifying this technically demanding method. PMID- 9210380 TI - A role for cell migration in the sexual transmission of HIV-1? AB - The issue of how human immunodeficiency virus-1 (HIV-1) enters the body following sexual contact has been the subject of considerable controversy. Several possible routes for the initial infection have been suggested [1-6], including the possibility that the transmission is mediated by HIV-1-infected lymphocytes or macrophages in serum and female genital tract secretions, rather than by free virus. We recently reported that HIV-1-infected, activated primary monocytes can migrate between epithelial cells grown in confluent monolayer cultures in vitro [7]. We report here on experiments carried out in mice to test the hypothesis that mononuclear blood cells are capable of migrating through intact epithelia, and thus of carrying a virus into an animal. We placed double-stained, activated mononuclear blood cells into the vaginas of mice; four hours later, numerous double-stained cells were observed in the connective tissue beneath the vaginal epithelium and the iliac lymph nodes of the experimental mice. We speculate that such migration may be involved in the sexual transmission of HIV-1. PMID- 9210383 TI - Colour vision. PMID- 9210384 TI - Immune surveillance and AIDS progression. PMID- 9210381 TI - S-phase function of Drosophila cyclin A and its downregulation in G1 phase. AB - BACKGROUND: Cyclin E is the normal inducer of S phase in G1 cells of Drosophila embryos. Stable G1 quiescence requires the downregulation both of cyclin E and of other factors that can bypass the normal regulation of cell cycle progression. RESULTS: High-level expression of cyclin A triggered the G1/S transition in wild type embryos and in mutant embryos lacking cyclin E. Three types of control downregulated this activity of cyclin A. First, cyclin destruction limited the accumulation of cyclin A protein in G1. Second, inhibitory phosphorylation of cdc2, the kinase partner of cyclin A, reduced the S-phase promoting activity of cyclin A in G1. Third, rux, a protein with unknown biochemical function, limited cyclin A function in G1. Overexpression of rux blocked S phase induction by coexpressed cyclin A and promoted the degradation of cyclin A. Rux also prevented a stable cyclin A mutant from inducing S phase, indicating that inhibition does not require cyclin destruction, and drove the nuclear localization of cyclin A. CONCLUSIONS: Cyclin A can drive the G1/S transition, but this function is suppressed by three types of control: cyclin A destruction, inhibitory phosphorylation of cdc2, and inhibition by rux. The partly redundant contributions of these three inhibitory mechanisms safeguard the stability of G1 quiescence until the induction of cyclin E. The action of rux during G1 resembles the action of inhibitors of mitotic kinases present during G1 in yeast, although no obvious sequence similarity exists. PMID- 9210390 TI - Environmental biotechnology. PMID- 9210391 TI - Web alert. Environmental biotechnology. Regulatory affairs. PMID- 9210393 TI - Mechanism-based inactivation of lacrimal-gland peroxidase by phenylhydrazine: a suicidal substrate to probe the active site. AB - Humans are exposed to various hydrazine derivatives for therapeutic control of several diseases, and mammalian peroxidases are implicated in the oxidative metabolism of many drugs. The results presented here indicate that lacrimal-gland peroxidase is irreversibly inactivated in a mechanism-based way by phenylhydrazine, which acts as a suicidal substrate in the presence of H2O2. The pseudo-first-order kinetic constants for inactivation at pH 5.5 are Ki=18 microM, kinact=0.25 min-1 and tau50=2.75 min, with a second-order rate constant of 0.75x10(4) M-1.min-1. Approx. 27 mol of phenylhydrazine and 54 mol of H2O2 are required per mol of enzyme for complete inactivation. The pH-dependent inactivation kinetics indicate the involvement of an ionizable group on the enzyme with a pKa value of 5.4, protonation of which favours inactivation. SCN-, the plausible physiological electron donor of the enzyme, protects it from inactivation. Binding studies by optical difference spectroscopy indicate that phenylhydrazine interacts with the enzyme with a KD value of 60 microM, and its binding is prevented by the presence of SCN-. The enzyme is also protected by 5, 5-dimethyl-1-pyrroline N-oxide, a free-radical trap, suggesting the involvement of a radical species in the inactivation. ESR studies indicate the formation of a spin-trapped phenyl radical (aN=15.9G and abetaH=24.8G) generated on incubation of phenylhydrazine with the enzyme and H2O2. A 75% loss of the Soret spectrum is observed when the enzyme is completely inactivated. However, in the presence of the spin trap, spectral loss is prevented and the enzyme compound II is readily reduced to the native state by phenylhydrazine. The phenylhydrazine-inactivated enzyme reacts with H2O2 or CN- to form compound II or the cyanide complex with a characteristic spectrum, indicating that haem iron is protected from attack by the radical species. The inactivated enzyme binds SCN- with a KD value similar to that of the native enzyme (15+/-3 mM), suggesting that the donor-binding site remains unaffected. CD studies of the inactive enzyme show complete disappearance of the Soret band at 409 nm with the appearance of a new band at 275 nm. This indicates that the haem environment of the enzyme is perturbed in the inactive form. As benzene, the end product of phenylhydrazine oxidation, has no effect on the enzyme, we suggest that the phenyl radical formed by one-electron oxidation by catalytically active enzyme inactivates it by incorporation in the vicinity of its haem moiety. The data support the use of phenylhydrazine as a probe for structural and mechanistic analysis of the active site of the lacrimal-gland peroxidase. PMID- 9210394 TI - Mode of inhibition of HIV reverse transcriptase by 2-hexaprenylhydroquinone, a novel general inhibitor of RNA-and DNA-directed DNA polymerases. AB - A natural compound from the Red Sea sponge Ircinia sp., 2-hexaprenylhydroquinone (HPH), has been shown to be a general inhibitor of retroviral reverse transcriptases (from HIV-1, HIV-2 and murine leukaemia virus) as well as of cellular DNA polymerases (Escherichia coli DNA polymerase I, and DNA polymerases alpha and beta). The pattern of inhibition was found to be similar for all DNA polymerases tested. Thus the mode of inhibition was studied in detail for HIV-1 reverse transcriptase. HPH is a non-competitive inhibitor and binds the enzyme irreversibly with high affinity (Ki=0. 62 microM). The polar hydroxy groups have been shown to be of key importance. A methylated derivative, mHPH, which is devoid of these polar moieties, showed a significantly decreased capacity to inhibit all DNA polymerases tested. Like the natural product, mHPH binds the enzyme independently at an allosteric site, but with reduced affinity (Ki=7.4 microM). We show that HPH does not interfere with the first step of the polymerization process, i.e. the physical formation of the reverse-transcriptase DNA complex. Consequently, we suggest that the natural inhibitor interferes with the subsequent steps of the overall reaction. Since HPH seems not to affect the affinity of dNTP for the enzyme (the Km is unchanged under conditions where the HPH concentration is increased), we speculate that its inhibitory capacity is derived from its effect on the nucleotidyl-transfer catalytic reaction. We suggest that such a mechanism of inhibition is typical of an inhibitor whose mode of inhibition should be common to all RNA- and DNA-directed polymerases. PMID- 9210392 TI - The vacuolar H+-ATPase: a universal proton pump of eukaryotes. AB - The vacuolar H+-ATPase (V-ATPase) is a universal component of eukaryotic organisms. It is present in the membranes of many organelles, where its proton pumping action creates the low intra-vacuolar pH found, for example, in lysosomes. In addition, there are a number of differentiated cell types that have V-ATPases on their surface that contribute to the physiological functions of these cells. The V-ATPase is a multi-subunit enzyme composed of a membrane sector and a cytosolic catalytic sector. It is related to the familiar FoF1 ATP synthase (F-ATPase), having the same basic architectural construction, and many of the subunits from the two display identity with one another. All the core subunits of the V-ATPase have now been identified and much is known about the assembly, regulation and pharmacology of the enzyme. Recent genetic analysis has shown the V-ATPase to be a vital component of higher eukaryotes. At least one of the subunits, i.e. subunit c (ductin), may have multifunctional roles in membrane transport, providing a possible pathway of communication between cells. The structure of the membrane sector is known in some detail, and it is possible to begin to suggest how proton pumping is coupled to ATP hydrolysis. PMID- 9210395 TI - Regulatory domains of the A-Myb transcription factor and its interaction with the CBP/p300 adaptor molecules. AB - The A-Myb transcription factor belongs to the Myb family of oncoproteins and is likely to be involved in the regulation of proliferation and/or differentiation of normal B cells and Burkitt's lymphoma cells. To characterize in detail the domains of A-Myb that regulate its function, we have generated a series of deletion mutants and have investigated their trans-activation potential as well as their DNA-binding activity. Our results have allowed us to delineate the trans activation domain as well as two separate regulatory regions. The boundaries of the trans-activation domain (amino acid residues 218-319) are centred on a sequence rich in charged amino acids (residues 259-281). A region (residues 320 482) localized immediately downstream of the trans-activation domain and containing a newly identified conserved stretch of 48 residues markedly inhibits specific DNA binding. Finally the last 110 residues of A-Myb (residues 643-752), which include a sequence conserved in all mammalian myb genes (region III), negatively regulate the maximal trans-activation potential of A-Myb. We have also investigated the functional interaction between A-Myb and the nuclear adaptor molecule CBP [cAMP response element-binding protein (CREB)-binding protein]. We demonstrate that CBP synergizes with A-Myb in a dose-dependent fashion, and that this co-operative effect can be inhibited by E1A and can also be observed with the CBP homologue p300. We show that this functional synergism requires the presence of the A-Myb charged sequence and that it involves physical interaction between A-Myb and the CREB-binding domain of CBP. PMID- 9210396 TI - Interferon alpha2 recombinant and epidermal growth factor modulate proliferation and hypusine synthesis in human epidermoid cancer KB cells. AB - We previously found that interferon alpha2 recombinant (IFNalpha) increases the expression of epidermal growth factor receptor (EGF-R) in the human epidermoid cancer KB cell line. Here we report the effects of IFNalpha and epidermal growth factor (EGF) on KB cell cycle kinetics. IFNalpha (1000 i.u./ml) for 48 h decreased the S-phase fraction and diminished the expression of Ki67 and proliferating cell nuclear antigen on KB cells. Incubation of IFNalpha-treated KB cells with 10 nM EGF for 12 h reversed these effects. We then studied several biochemical markers of cell proliferation. Ornithine decarboxylase activity was decreased to about one-tenth by IFNalpha and partly restored by EGF. Hypusine is contained only in eukaryotic initiation factor 5A and its levels are correlated with cell proliferation. IFNalpha decreased hypusine synthesis by 75%; exposure of cells to EGF for 12 h restored hypusine synthesis almost completely. We also studied the effects of IFNalpha on the cytotoxicity of the recombinant toxin TP40, which inhibits elongation factor 2 through EGF-R binding and internalization. IFNalpha greatly enhanced the TP40-induced inhibition of protein synthesis in KB cells. In conclusion, IFNalpha, which affects protein synthesis machinery and increases EGF-R expression, enhances the tumoricidal activity of TP40 and hence could be useful in the setting of anti-cancer therapy. PMID- 9210397 TI - Comparison of the ligand-binding properties of native and copper-less cytochromes bo from Escherichia coli. AB - The binding of four anionic ligands, cyanide, fluoride, azide and formate, to cytochrome bo purified from Escherichia coli cells grown with a copper supplement (+Cu cyt.bo) is described. Membrane-bound cytochrome bo that lacks the copper component, CuB, of its active site can be prepared from cells grown under conditions where the availability of copper is limited by the presence of a CuI chelator, 2,2'-bicinchinonic acid. The ligand-binding properties of this copper less enzyme (-Cu cyt.bo) are compared with those of +Cu cyt. bo. As judged from near-UV/visible spectroscopic changes, cyanide forms a low-spin complex with +Cu cyt.bo, whereas azide, fluoride and formate form high-spin complexes. The pH dependences of binding suggest that for all four of these anionic ligands, both the rates of binding and the binding affinities are primarily dependent on the concentration of their protonated forms. -Cu cyt.bo, which shows less than 15% of the duroquinol oxidase activity of +Cu cyt.bo, binds cyanide, azide and fluoride, but with greatly decreased affinity (<1/30, 1/2000 and 1/2500 respectively at pH5.5 compared with +Cu cyt.bo). The complex of azide with -Cu cyt.bo still seems to be high-spin and azide binding to -Cu cyt.bo is still pH-dependent, although less so than azide binding to +Cu cyt.bo. PMID- 9210398 TI - Age-related changes in effects of insulin-like growth factor I on human osteoblast-like cells. AB - The role of insulin-like growth factor I (IGF-I) in extracellular matrix metabolism was studied in both proliferating and confluent human osteoblast-like cultures derived from donors of different ages. In proliferating cultures, recombinant human (rh)IGF-I was found to increase the incorporation of [3H]thymidine in a dose- and age-dependent manner. To study cell proliferation dynamically, continuous growth curves with and without rhIGF-I were modelled by a modified logistic function. Increasing doses of rhIGF-I decreased the lag time and maximal growth rates, whereas plateau values decreased only at the highest dose (100 ng/ml). In post-proliferative cell strains, rhIGF-I (0.1-100 ng/ml) increased levels of type I collagen, biglycan and decorin, and to a smaller extent fibronectin and thrombospondin, whereas it decreased the levels of hyaluronan and a versican-like proteoglycan when protein and proteoglycan metabolism were followed by steady-state radiolabelling with [3H]proline, [3H]glucosamine or [35S]sulphate. These responses to rhIGF-I were found to be age dependent, with osteoblast-like cells derived from younger patients being more responsive to rhIGF-I. When extracellular matrix turnover was analysed by pulse chase experiments, rhIGF-I had no effect. The steady-state levels of collagen, decorin, hyaluronan and a versican-like proteoglycan for bone cells treated with rhIGF-I on day 7 in culture were equivalent to levels of these matrix components in untreated osteoblasts grown for 14 days. These results are consistent with rhIGF-I's altering cellular proliferative capacity and matrix synthesis, causing a change in the osteoblast differentiated state. PMID- 9210400 TI - Mechanism of apoptotic cell death of human gastric carcinoma cells mediated by transforming growth factor beta. AB - Human gastric carcinoma cell line HSC-39 has been shown to undergo apoptotic cell death in response to treatment with transforming growth factor beta1 (TGF-beta1). To understand better the cell death mechanism in this TGF-beta1-mediated apoptosis, we investigated the effect of the expression of TGF-beta-stimulated clone 22 (TSC-22) on cell death events. TGF-beta1 induced TSC-22 gene expression in HSC-39 cells only when the cells had previously been adapted to the serum-free culture conditions required to undergo TGF-beta1-mediated apoptosis. HSC-39 cells transfected with a TSC-22 expression vector showed a significant decrease in cell viability compared with those transfected with a control vector. The cellular events characteristic of apoptosis, chromatin condensation and DNA fragmentation were observed only in cells transfected with a TSC-22 expression vector. On immunostaining of the transfected cells, almost every cell that expressed TSC-22 tagged with influenza virus haemagglutinin exhibited the morphology of an apoptotic cell. Partial protection from the cell death effect of TGF-beta1 on HSC 39 cells was observed when cells were treated with acetyl-L-aspartyl-L-glutamyl-L valyl-L-aspart-1-al (Ac-DEVD-CHO, an inhibitor specific for CPP32-type protease). Protection against cell death by the transfection of a TSC-22 expression vector was also offered by Ac-DEVD-CHO addition. These results suggest that TSC-22 elicits the apoptotic cell death of human gastric carcinoma cells through the activation of CPP32-like protease and mediates the TGF-beta1 signalling pathway to apoptosis. PMID- 9210399 TI - Validation and steady-state analysis of a power-law model of purine metabolism in man. AB - The paper introduces a model of human purine metabolism in situ. Chosen from among several alternative system descriptions, the model is formulated as a Generalized Mass Action system within Biochemical Systems Theory and validated with analyses of steady-state and dynamic characteristics. Eigenvalue and sensitivity analyses indicate that the model has a stable and robust steady state. The model quite accurately reproduces numerous biochemical and clinical observations in healthy subjects as well as in patients with disorders of purine metabolism. These results suggest that the model can be used to assess biochemical and clinical aspects of human purine metabolism. It provides a means of exploring effects of enzyme deficiencies and is a potential tool for identifying steps of the pathway that could be the target of therapeutical intervention. Numerous quantitative comparisons with data are given. The model can be used for biomathematical exploration of relationships between enzymic deficiencies and clinically manifested diseases. PMID- 9210401 TI - Inhibition of the expression of ornithine decarboxylase and c-Myc by cell permeant ceramide in difluoromethylornithine-resistant leukaemia cells. AB - Ceramide has emerged as a novel lipid mediator in cell growth and apoptosis. In difluoromethylornithine-resistant L1210 cells stimulated to growth from quiescence, the cell-permeant analogues of ceramide N-acetylsphingosine (C2 ceramide) and N-hexanoylsphingosine (C6-ceramide) inhibited the induction of ornithine decarboxylase (ODC) activity with IC50 of 8.3 and 1.5 microM respectively. This effect was strictly related to the ability to inhibit cell growth and [3H]thymidine incorporation. The suppression of cell growth was also associated with apoptosis. The addition of bacterial sphingomyelinase resulted in a significant, but limited, reduction of ODC induction and [3H]thymidine incorporation. Bacterial lipopolysaccharide, which may act as a ceramide analogue, also inhibited the induction of the enzyme. Moreover, C6-ceramide largely prevented the accumulation of ODC mRNA and its precursor, ODC heterogeneous nuclear RNA, that accompanied the induction of ODC activity. A slight increase in ODC turnover was also observed. The DNA-binding activity of some transcription factors known to bind and transactivate the ODC gene was investigated by gel mobility-shift assay under the same experimental conditions. However, only the binding of Myc/Max was negatively affected by the treatment with C6-ceramide. Furthermore, the amount of immunoreactive c-Myc, which increased after stimulation of the cells to growth, was strongly reduced by C6 ceramide. These results suggest that the inhibition of c-Myc and ODC expression may be early events in the response of leukaemia cells to ceramide. PMID- 9210402 TI - Evidence for glutathione involvement in platelet-derived growth-factor-mediated signal transduction. AB - Recent studies show that glutathione, while being involved in the well-known physiological processes of amino acid transport and detoxification, can also play a part in cell proliferation events. Cell treatment with l-buthionine sulphoximine, which causes glutathione depletion, is accompanied by a decrease in cell proliferation. At present no precise relationship between this thiol and any critical intermediate of the mitogenic cascade has been proved. In this study, conducted on NIH/3T3 murine fibroblasts, we demonstrate a strict correlation between glutathione levels and platelet-derived growth-factor-receptor activation in response to stimulation and cell proliferation. The receptor autophosphorylation is severely impaired at low glutathione cellular levels. The interaction of glutathione with this growth-factor receptor in vivo, while being rather specific, is complex and may involve both cytosolic and extracellular receptor domains. PMID- 9210404 TI - Structure, organization and expression of the mouse ornithine decarboxylase antizyme gene. AB - Ornithine decarboxylase antizyme is a protein that participates in the regulation of cellular polyamine levels. In this study we have isolated and sequenced the mouse gene encoding antizyme protein. Transfection of various cell lines with a 5.5 kb genomic fragment containing the antizyme locus resulted in the production of a 29 kDa antizyme protein, confirming that this locus contained a functional gene. Comparison of the mouse gene with the corresponding rat gene [Miyazaki, Matsufuji and Hayashi, (1992) Gene 113, 191-197] revealed an identical exon/intron organization and high level of nucleotide sequence conservation that was 89% for the entire transcription unit. Protein-coding regions of the two genes exhibited 97% nucleotide sequence identity and there were only four amino acid differences between the 227-residue antizyme protein sequences of the mouse and rat. The promoter of the antizyme gene was functional in mouse (N2A and NIH/3T3) and hamster (CHO) cell lines. The presence of 0.1 mM spermidine in culture medium increased the amount of immunoreactive antizyme protein in cells transfected with the antizyme gene or antizyme cDNA, possibly owing to facilitated frameshifting in the translation of antizyme mRNA. Recombinant antizyme protein was also produced in Escherichia coli and used to raise specific polyclonal antibodies in rabbits and to devise immunological methods for the measurement of antizyme concentration. PMID- 9210403 TI - Tridegin, a new peptidic inhibitor of factor XIIIa, from the blood-sucking leech Haementeria ghilianii. AB - 1. Crude salivary gland extract of the giant Amazon leech, Haementeria ghilianii, contains an inhibitor of plasma factor XIIIa. 2. The inhibitory agent was purified to homogeneity by anion-exchange, cation-exchange, gel-filtration and reverse-phase chromatography to yield a single band on SDS/PAGE with an apparent molecular mass of 7.3 kDa. It has been named tridegin. 3. Micro-sequencing of proteolytic fragments showed tridegin to be a peptide of 66 amino acids. The sequence is unique with little similarity to other leech-derived proteins. 4. Inhibition of plasma factor XIIIa activity was confirmed by four independent methods: tridegin increased the solubility of fibrin clots in urea, inhibited ammonia produced from the incorporation of ethylamine into casein, inhibited the incorporation of 5'-(biotinamido)pentylamine into casein and prevented gamma dimer formation in clotting fibrinogen. 5. The IC50 of tridegin (approx. 9.2 nM) is very close to the concentration of factor XIIIa used in the assay and in fact depends on its concentration. This is the most potent inhibitor of factor XIIIa yet described. 6. Tridegin also inhibits platelet factor XIIIa (factor XIIIAa) with a similar potency to that of the plasma enzyme. 7. Tridegin also inhibits tissue transglutaminase but with lower potency and independently of the enzyme concentration. 8. Tridegin appears to be specific for transglutaminases, since it has no effect on the coagulation times of human plasma, on thrombin or factor Xa. Moreover it has no effect on other thiol-containing enzymes and has no ability to digest fibrinogen or cleave the isopeptide substrate, L-gamma-glutamyl-4 nitroanilide. PMID- 9210406 TI - Stimulation of transcription in vitro from a liver-specific promoter by human glucocorticoid receptor (hGRalpha). AB - The rat tyrosine aminotransferase (TAT) gene is a liver-specific and glucocorticoid-inducible gene. Previous studies have shown that the TAT promoter (TAT0.35; nt -350 to +1) is able to sustain liver-specific gene expression both in transient transfection and in a transcription assay in vitro [Schweizer Groyer, Groyer, Cadepond, Grange, Baulieu and Pictet (1994) Nucleic Acids Res. 22, 1583-1592]. Here we report that the basal transcriptional activity generated from TAT0.35 in the presence of crude liver nuclear extracts is enhanced by added human glucocorticoid receptor (hGRalpha), provided that TAT0.35 sequences were flanked (5') with a glucocorticoid responsive unit (GREII of the TAT gene, including its 5'-CCAAT flanking sequence). Two sources of hGRalpha were used: nuclear extracts prepared from Sf9 insect (Sf9-NEs) cells over-expressing hGRalpha, and hGRalpha from pRShGRalpha-transfected COS-7 cells, enriched by high performance ion-exchange chromatography. The enhancement of transcription in vitro (1.5-4.5-fold) was dependent on the amount of added hGRalpha and independent of the nature (agonist or antagonist) of the ligand. Moreover, the hGRalpha-mediated stimulation of transcription was (i) dependent on GRE/progesterone response element (PRE) (it was inhibited by a 25-fold excess of GRE/PRE but not by a 100-fold excess of oestrogen response element) and (ii) receptor-dependent (Sf9-NEs prepared from uninfected Sf9 cells or from Sf9 cells infected with wild-type baculoviral DNA did not enhance transcription). Taken together, these experiments support the conclusions that in vitro the glucocorticoid receptor is able to enhance transcription from genomic, liver specific, promoter sequences (those of the TAT gene), and that this enhancement of transcription from the liver-specific TAT0.35 promoter is dependent both on the glucocorticoid receptor and on the latter's interaction with its cognate response elements. PMID- 9210405 TI - Construction, expression and characterization of chimaeric toxins containing the ribonucleolytic toxin restrictocin: intracellular mechanism of action. AB - Restrictocin is a ribonucleolytic toxin produced by the fungus Aspergillus restrictus. Two chimaeric toxins containing restrictocin directed at the human transferrin receptor have been constructed. Anti-TFR(scFv)-restrictocin is encoded by a gene produced by fusing the DNA encoding a single-chain antigen combining region (scFv) of a monoclonal antibody, directed at the human transferrin receptor, at the 5' end of that encoding restrictocin. The other chimaeric toxin, restrictocin-anti-TFR(scFv), is encoded by a gene fusion containing the DNA encoding the single-chain antigen-combining region of antibody to human transferrin receptor at the 3' end of the DNA encoding restrictocin. These gene fusions were expressed in Escherichia coli, and fusion proteins purified from the inclusion bodies by simple chromatography techniques to near homogeneity. The two chimaeric toxins were found to be equally active in inhibiting protein synthesis in a cell-free in vitro translation assay system. The chimaeric toxins were selectively toxic to the target cells in culture with potent cytotoxic activities. However, restrictocin-anti-TFR(scFv) was more active than anti-TFR(scFv)-restrictocin on all cell lines studied. By using protease and metabolic inhibitors, it can be shown that, to manifest their cytotoxic activity, the restrictocin-containing chimaeric toxins need to be proteolytically processed intracellularly and the free toxin or a fragment thereof thus generated is translocated to the target via a route involving the Golgi apparatus. PMID- 9210407 TI - Studies of cellulose binding by cellobiose dehydrogenase and a comparison with cellobiohydrolase 1. AB - The binding isotherm to cellulose of cellobiose dehydrogenase (CDH) from Phanerochaete chrysosporium has been compared with that of cellobiohydrolase 1 (CBH 1) from Trichoderma reesei. CDH binds more strongly but more sparsely to cellulose than does CBH 1. In a classical Scatchard analysis, a better fit to a one-site binding model was obtained for CDH than for CBH 1. The binding of both enzymes decreased in the presence of ethylene glycol, increased in the presence of ammonium sulphate and was unaffected by sodium chloride. Attempts to localize the cellulose-binding site on CDH have also been made by exposing enzymically digested CDH to cellulose and isolating the cellulose-bound peptides. The results suggest that the cellulose-binding site is located internally in the amino acid sequence of CDH. PMID- 9210408 TI - Insulin stimulates tyrosine phosphorylation of the proto-oncogene product of c Cbl in 3T3-L1 adipocytes. AB - We report here that the product of the c-Cbl proto-oncogene is prominently tyrosine phosphorylated in response to insulin in 3T3-L1 adipocytes. The tyrosine phosphorylation of c-Cbl reaches a maximum within 1-2 min after stimulation by insulin and gradually declines thereafter. The tyrosine phosphorylation of c-Cbl was also observed after treatment of 3T3-L1 adipocytes with epidermal growth factor, whereas platelet-derived growth factor had no effect. After insulin dependent tyrosine phosphorylation, c-Cbl specifically associates with fusion proteins containing the Src homology 2 (SH2) domains of Crk and the Fyn tyrosine kinase, but not with fusion proteins containing the SH2 domains of either the p85 subunit of phosphatidylinositol 3'-kinase or the tyrosine phosphatase SHPTP2/Syp. Furthermore insulin stimulates the association of c-Cbl with endogenous c-Crk and Fyn in intact 3T3-L1 adipocytes. The tyrosine phosphorylation of c-Cbl is regulated during adipocyte differentiation. Neither insulin-like growth factor 1 nor insulin stimulated the tyrosine phosphorylation of c-Cbl in 3T3-L1 fibroblasts. Moreover, c-Cbl is not tyrosine phosphorylated in response to insulin in cells expressing high levels of the human insulin receptor, or in hepatocytes, despite comparable levels of c-Cbl expression. These results suggest that c-Cbl might have a novel function in the regulation of insulin receptor intracellular signalling in 3T3-L1 adipocytes. PMID- 9210409 TI - Alpha-difluoromethylornithine-resistant cell lines obtained after one-step selection of Leishmania mexicana promastigote cultures. AB - Proliferation of Leishmania mexicana promastigotes in synthetic medium can be blocked by the depletion of intracellular polyamine pools induced by the presence of D,L-alpha-difluoromethylornithine (DFMO), a specific and irreversible inhibitor of ornithine decarboxylase (ODC). Here we report that DFMO-resistant cell lines growing normally at DFMO levels of 10 mM have been obtained from non proliferating cultures after a single-step selection in the presence of high concentrations of the drug. The DFMO-resistant promastigotes underwent a morphological transformation into an 'amastigote-like' form after incubation for several hours at gradually increasing temperatures up to 35 degrees C. The uptake of DFMO was not significantly altered in the drug-resistant cell lines but in both cases (promastigote and 'amastigote-like' forms) the ODC specific activity was increased approx. 15-fold over the normal enzymic levels found in the wild type Leishmania. The enzyme affinities for its substrate and for DFMO gave very similar values in the drug-resistant promastigotes and the wild-type parasites. In contrast, ODC from the 'amastigote-like' Leishmania showed a higher affinity for ornithine and a decreased capacity for the binding of DFMO. An 80-fold amplification of the ODC gene and a corresponding increase in its transcripts have been detected in both DFMO-resistant Leishmania cell lines. The drug resistant phenotypes with their characteristic morphologies, the increased levels of ODC activity and the amplification of the ODC gene have been stable for at least 6 months in the absence of selective pressure. PMID- 9210410 TI - Receptor dimerization is not a factor in the signalling activity of a transforming variant epidermal growth factor receptor (EGFRvIII). AB - The type-III deletion variant of the epidermal growth factor receptor (EGFRvIII) is frequently found in glioblastomas and other malignant human tumours. Although EGFRvIII confers ligand-independent oncogenic transformation of cell lines, the mechanism by which it promotes aberrant cellular proliferation is unknown. Using cell lines expressing comparable numbers of either wild-type receptor (EGFRwt) or EGFRvIII, we compared several parameters of receptor activation: dimerization, tyrosine phosphorylation and activation of intracellular signalling proteins. Like activated EGFRwt, EGFRvIII was phosphorylated and bound constitutively to the Shc adapter protein. Indeed, EGFRvIII-associated Shc had a higher phosphotyrosine content than Shc associated with stimulated EGFRwt. EGFRwt dimerized in response to either EGF or transforming growth factor alpha. Higher cross-linker concentrations and incubation at higher temperatures (37 degrees C) allowed detection of EGFRwt dimers even in the absence of exogenous ligand. In contrast, EGFRvIII failed to dimerize under any conditions studied. Moreover, neither mitogen-activated protein kinase nor phospholipase Cgamma were phosphorylated in EGFRvIII-expressing cells. We conclude that the deletion of 267 amino acids from the 621-amino-acid N-terminal domain of EGFR does not result simply in a constitutively activated receptor, but alters the spectrum of signalling cascades utilized. Furthermore the ligand-independent transforming activity of EGFRvIII is independent of receptor dimerization. PMID- 9210411 TI - Identification of a minimal promoter element of the mouse epidermal growth factor gene. AB - We have previously generated a transgenic mouse line (EGF/Tag) in which simian virus 40 (SV40) T-antigen expression is directed by the mouse epidermal growth factor (EGF) gene promoter. In these mice, cellular hyperproliferation is observed in the submaxillary gland associated with SV40 T-antigen expression. In addition, SV40 T-antigen-expressing tumours of prostatic origin are seen. We have now derived immortalized cell lines from these tissues and have used the cells to perform a functional analysis of the EGF gene promoter. Cells were transfected with EGF promoter/reporter constructs, and an element located between 51 and 35 bases upstream of the EGF mRNA start site required for basal activity of the promoter was identified. Electrophoretic mobility-shift analysis suggests that three proteins bind to this region, one of which is either Sp1 or a closely related protein. PMID- 9210412 TI - Specificity and kinetic effects of nitrophenol analogues that activate myosin subfragment 1. AB - 2,4-Dinitrophenol (DNP) activates the myosin ATPase of mammalian skeletal muscle in the presence of Ca2+ or Mg2+, and inhibits it when the bivalent cations are replaced by K+ and EDTA. Activation of Mg2+ATPase is abolished by the presence of unregulated actin. 3-Nitrophenol (3-NP) is also an activator, whereas other analogues (2-nitrophenol, 2-NP, and 4-nitrophenol, 4-NP) are much less effective. Concentrations required for their half-maximal effects (K0.5) range from 2 to 15 mM for 3-NP and DNP in the presence of different cations, and the sequence for the analogues is 3-NP<=DNP<<2-NP approximately 4-NP, which is apparently unrelated to either hydrophobicity or pK. DNP and 3-NP have almost identical effects on the ATPase activity of chymotryptic subfragment 1 as they do on myosin, which is an indication that their target is the globular head region rather than the tail, or the 18 kDa (regulatory) light chain. Analysis of the ATP concentration dependence for subfragment- 1 ATPase in the presence of Ca2+ or Mg2+ shows that DNP activates only at high substrate concentrations, becoming increasingly effective with ATP concentrations in the physiological range. At low substrate concentrations, DNP inhibits hydrolysis by increasing the apparent Km for ATP at the catalytic site. In the presence of Mg2+, it mimics the effect of actin, which increases the Km and accelerates the release of products following hydrolysis. At high substrate concentrations, activation by DNP appears to involve a kinetic component with low affinity for ATP that can increase the overall reaction rate by a factor of 2- to 9-fold, depending on the bivalent cation. This low-affinity component is either induced by the drug (in the presence of Mg2+) or shifted by the drug to a lower ATP concentration range (in the presence of Ca2+). PMID- 9210414 TI - 1-O-Octadecyl-2-O-methylglycerophosphocholine inhibits protein kinase C-dependent phosphorylation of endogenous proteins in MCF-7 cells. AB - Studies with leukaemic cells, based primarily on in vitro assays, have suggested that antitumour ether lipids have only a moderate effect on protein kinase C (PKC) activity, and, furthermore, inhibition of PKC is unlikely to be involved in the mechanism of inhibition of cell proliferation by these compounds. To determine if this is also the case for epithelial cancer cells, we examined the effect of 1-O-octadecyl-2-O-methylglycerophosphocholine (ET18-OCH3) on PKC induced phosphorylation of endogenous proteins in MCF-7 cells under incubation conditions where the drug inhibited cell proliferation. As expected, stimulation of quiescent 32P-labelled MCF-7 cells with 1 microM PMA resulted in the phosphorylation of a number of proteins. The PMA-induced phosphorylation of the proteins was abolished by preincubation of the cells with Ro 31-8220 (5 microM) for 20 min, or 10 microg/ml ET18-OCH3 for 3 h before stimulation with PMA. Thus under incubation conditions where ET18-OCH3 inhibited the proliferation of MCF-7 cells, the ether lipid potently inhibited the activity of PKC in intact cells. This inhibition was unlikely to be due to the effect of the compound on PKC translocation since there was little effect of ET18-OCH3 on the translocation of the alpha, gamma and epsilon species of PKC. These results suggest that a role for the inhibition of PKC activity by ET18-OCH3 in the mechanism of inhibition of cell proliferation by ET18-OCH3 cannot yet be discounted in epithelial cancer cells. In addition, we also observed that ET18-OCH3 enhanced the phosphorylation of selected proteins under basal unstimulated conditions. Although some of these proteins were also observed to be phosphorylated in response to PMA stimulation, the phosphorylation induced by ET18-OCH3 was not inhibited by Ro 31-8220, indicating that this was not mediated by PKC. PMID- 9210413 TI - Expression of a variant surface glycoprotein of Trypanosoma gambiense in procyclic forms of Trypanosoma brucei shows that the cell type dictates the nature of the glycosylphosphatidylinositol membrane anchor attached to the glycoprotein. AB - Procyclic forms of Trypanosoma brucei have been genetically modified to express the major metacyclic variant surface glycoprotein (VSG variant AnTat 11.17) of Trypanosoma gambiense. The VSG is expressed in an intact membrane-bound form that can be detected over the entire plasma membrane, together with procyclin, and as a series of lower-molecular-mass fragments that are mostly soluble degradation products. The presence of degraded VSG in the cells and the culture medium suggests that VSG is not efficiently processed and/or efficiently folded when expressed in procyclic cells. The level of procyclin expressed on the surface of these cells is slightly reduced, although there is no difference in procyclin mRNA levels. The intact membrane-bound form of the VSG is N-glycosylated with oligomannose structures and contains a glycosylphosphatidylinositol (GPI) membrane anchor that can be biosynthetically labelled with [3H]ethanolamine. The anchor is sensitive to mammalian GPI-specific phospholipase D but, like the anchor of procyclin, it is resistant to the action of bacterial phosphatidylinositol-specific phospholipase C. This pattern of phospholipase sensitivity suggests that the GPI anchor acquired by VSG when expressed in procyclics is acylated on the inositol ring and therefore resembles a procyclic procyclin-type anchor rather than a trypomastigote VSG-type anchor with respect to the lipid structure. The VSG expressed in procyclics was sensitive to the action of a mixture of sialidase, beta-galactosidase and beta-hexosaminidase, suggesting that the VSG GPI anchor also contains a sialylated polylactosamine side-chain modification similar to that described for procyclin. These results indicate that the nature of the protein expressed has little influence on the post-translational modifications performed in the secretory pathway of procyclic trypanosomes. PMID- 9210415 TI - Plasmodium falciparum CTP:phosphocholine cytidylyltransferase expressed in Escherichia coli: purification, characterization and lipid regulation. AB - The Plasmodium falciparum CTP:phosphocholine cytidylyltransferase (PfCCT) has been isolated from an overexpressing strain of Escherichia coli. The plasmid pETPfCCT mediated the overexpression of the full-length polypeptide directly. The recombinant protein corresponded to 6-9% of the total cellular proteins and was found essentially in the insoluble membrane fraction. Urea at 6 M was used to solubilize the recombinant protein from the insoluble fraction. The CCT activity was restored upon the removal of urea, and the protein was subsequently purified to homogeneity on a Q-Sepharose column. Approx. 1.4 mg of pure enzyme was obtained from a 250 ml culture of E. coli. Biochemical properties, including in vitro substrate specificity and enzymic characterization, were assessed. The lipid regulation of the recombinant plasmodial CCT activity was characterized for the first time. The Km values were 0.49+/-0.03 mM (mean+/-S.E.M.) for phosphocholine and 10.9+/-0.5 mM for CTP in the presence of lipid activators (oleic acid/egg phosphatidylcholine vesicles), whereas the Km values were 0.66+/ 0.07 mM for phosphocholine and 28.9+/-0.8 mM for CTP in the absence of lipid activators. The PfCCT activity was stimulated to the same extent in response to egg phosphatidylcholine vesicles containing anionic lipids, such as oleic acid, cardiolipin and phosphatidylglycerol, and was insensitive or slightly sensitive to PC vesicles containing neutral lipids, such as diacylglycerol and monoacylglycerol. Furthermore, the stimulated enzyme activity by oleic acid was antagonized by the cationic aminolipid sphingosine. These lipid-dependence properties place the parasite enzyme intermediately between the mammalian enzymes and the yeast enzyme. PMID- 9210416 TI - Selective release of human adipocyte fatty acids according to molecular structure. AB - The objective of the present study was to investigate the mobilization of individual fatty acids from human white fat cells. Mammary adipose tissue from eight healthy non-obese women in their normal dietary state was collected, and isolated adipocytes were incubated with lipolytic agents. The mobilization of 34 individual fatty acids was measured by comparing the composition of non esterified fatty acids (NEFA) with that of the triacylglycerols (TAG) from which they originated through lipolysis. Compared with TAG, NEFA were enriched in some polyunsaturated fatty acids with 18-20 carbon atoms. Conversely, the percentage of very-long-chain (20-22 carbon atoms) saturated and monounsaturated fatty acids was approx. 2 times lower in NEFA than in TAG. The relative mobilization (% in NEFA/% in TAG) of the most readily mobilized fatty acid (C20:5, n-3; 2.25) was more than 6-fold higher than that of the least readily mobilized (C22:1,n-11; 0.37). Relationships were found between the molecular structure of fatty acids and their mobilization rate. For a given chain length, the relative mobilization rate increased with increasing unsaturation, whereas for a given unsaturation, it decreased with increasing chain length. The relative mobilization rate for essential fatty acids decreased in the following order: C20:5,n-3>C20:4,n 6>C18:3,n-3>C18:2, n-6>C22:6,n-3. Interestingly, C20:5,n-3 and C20:4,n-6, which are respectively precursors of the 3- and 2-series of prostaglandins, were preferentially mobilized. It is concluded that fatty acids are selectively mobilized from human fat cells according to molecular structure, in full agreement with animal studies. By modulating the qualitative fatty acid supply to organs and by remodelling the fatty acid composition of adipose tissue, this selectivity would be relevant for consideration in physiology, health and epidemiology. PMID- 9210417 TI - Members of the nuclear factor 1 family and hepatocyte nuclear factor 4 bind to overlapping sequences of the L-II element on the rat pyruvate kinase L gene promoter and regulate its expression. AB - The L-II element (-149 to -126 bp) in the enhancer unit of the rat pyruvate kinase L (PKL) gene is required for cell-type-specific transcription and induction by carbohydrates. This element was found to bind multiple nuclear proteins with different heat stabilities. A heat-labile factor was shown to be hepatocyte nuclear factor (HNF) 4 by the electrophoretic mobility-shift assay (EMSA) using various competitor DNAs and anti-HNF4 serum. A heat-stable factor was purified from rat liver nuclear extract and was resolved as two protein bands migrating at about 33 kDa on SDS/polyacrylamide gels. Peptide sequence analysis revealed that these proteins were nuclear factor (NF) 1-L and NF1/Red1. The heat stable factor was also identified as a member of the NF1 family by using various competitor DNAs and anti-NF1 serum in an EMSA. In addition, we found that a factor bound to the accessory site of the rat S14 gene, which is necessary for carbohydrate responsiveness of this gene, was also a member of the NF1 family, raising the possibility that the NF1 family is involved in the carbohydrate regulation of gene transcription by interactions with other proteins. The NF1 family members and HNF4 interacted with overlapping sequences of the L-II element, wherein the 5' half-site was more critical for NF1 binding, and the 3' site was more important for HNF4 binding. Co-transfection of a vector expressing either NF1-L or NF1/Red1 repressed the transcription of the PKL enhancer unit chloramphenicol acetyltransferase (CAT) fusion gene in HepG2 cells, whereas co transfection of a vector expressing HNF4 activated the transcription of the same reporter gene. Furthermore NF1 family members antagonized the effect of HNF4 on PKL enhancer unit-CAT fusion gene expression when both expression plasmids were co-transfected. We conclude that NF1 family members and HNF4 regulate transcription of the PKL gene in an opposing manner by binding overlapping sequences of the L-II element. PMID- 9210418 TI - Gene structure and functional analysis of the human Na+/phosphate co-transporter. AB - Three lambda phage clones encompassing the Na+/phosphate co-transporter (NaPi-3) gene and its 5' flanking region were isolated from a human genomic DNA library. The gene comprises 13 exons and 12 introns and spans approx. 14 kb. All exon intron junctions conform to the GT/AG rule. The major transcription-initiation site was determined by primer-extension analysis and is an adenosine residue 57 bp upstream of the 3' end of the first exon. There is a typical TATA box 28 bp upstream of the major transcription-initiation site and various cis-acting elements, including a cAMP-responsive element, AP-1, AP-2 and SP-1 sites in the 5' flanking region. This region also contains three direct-repeat-like sequences that resemble the consensus binding sequence for members of the steroid-thyroid hormone receptor superfamily, including vitamin D. Deletion analysis suggests that the region from nt-2409 to nt-1259 in the 5' flanking region may be involved in kidney-specific gene expression. Vitamin D responsiveness of the NaPi-3 promoter was also detected in COS-7 cells co-transfected with a human vitamin D receptor expression vector. The presence of the three vitamin D receptor- responsive elements in the NaPi-3 promoter may be important in mediating the enhanced expression of the gene by 1,25-dihydroxyvitamin D3. PMID- 9210419 TI - Modulation of cathepsin D activity in retinal pigment epithelial cells. AB - This project used retinal pigment epithelial (RPE) cells to investigate the effects of up- and down-regulation of cathepsin D expression on the processing of cathepsin D and on the normal phagocytic and digestive function of these cells. RPE cells were transfected with a pHbetaApr-1-neo vector construct carrying the full-length sequence of the translated region of human cathepsin D in sense and antisense directions. Transfected cells were characterized for the presence and expression of the transgene by PCR amplification using transgene-specific primers. Total aspartic proteinase activity present in transformed RPE cells was measured by an enzyme assay using haemoglobin as substrate. Flow cytometry was used to quantify phagocytosis of fluorescein isothiocyanate-labelled rod outer segments (ROS), and lysosomal digestion of ROS was monitored by immunofluorescence. A 435 bp fragment was present in RPE cells carrying the cathepsin D transgene in sense and antisense orientations after PCR amplification. Expression of both 52 kDa procathepsin D and 34 kDa active cathepsin D was significantly up-regulated in sense cathepsin D-transfected RPE cells and down-regulated in RPE cells transfected with antisense cathepsin D. No other forms of cathepsin D were detected in the transfected cells, suggesting that, if pseudo-cathepsin D exists in RPE cells in vivo, it requires the presence of unknown specific regulatory elements. The up- and down-regulation of cathepsin D expression was further confirmed by enzyme assay. Transfected cells retained their phagocytosing ability after ROS challenge and maintained their ability to process ROS. The processing of ROS was significantly slower in RPE cells transfected with antisense than control vector or in sense-cathepsin D transfected cells. These results demonstrate that cathepsin D is a major proteolytic enzyme participating in the lysosomal digestion of photoreceptor outer segments. PMID- 9210420 TI - Characterization and purification of neutrophil ecto-phosphatidic acid phosphohydrolase. AB - Phosphatidic acid and its derivatives play potentially important roles as extracellular messengers in biological systems. An ecto-phosphatidic acid phosphohydrolase (ecto-PAPase) has been identified which effectively regulates neutrophil responses to exogenous phosphatidic acid by converting the substrate to diacylglycerol. The present study was undertaken to characterize this ecto enzyme on intact cells and to isolate the enzyme from solubilized neutrophil extracts. In the absence of detergent, short chain phosphatidic acids were hydrolysed most effectively by neutrophil plasma membrane ecto-PAPase; both saturated and unsaturated long chain phosphatidic acids were relatively resistant to hydrolysis. Both long (C18:1) and short (C8) chain lyso-phosphatidic acids were hydrolysed at rates comparable with those observed for short chain (diC8) phosphatidic acid. Activity of the ecto-enzyme accounted for essentially all of the N-ethylmaleimide-insensitive, Mg2+-independent PAPase activity recovered from disrupted neutrophils. At 37 degrees C and pH7.2, the apparent Km for dioctanoyl phosphatidic acid (diC8PA) was 1. 4x10(-3) M. Other phosphatidic acids and lysophosphatidic acids inhibited hydrolysis of [32P]diC8PA in a rank order that correlated with competitor solubility, lysophosphatidic acids and unsaturated phosphatidic acids being much more effective inhibitors than long chain saturated phosphatidic acids. Dioleoyl (C18:1) phosphatidic acid was an unexpectedly strong inhibitor of activity, in comparison with its ability to act as a direct substrate in the absence of detergent. Other inhibitors of neutrophil ecto-PAPase included sphingosine, dimethyl- and dihydro-sphingosine, propranolol, NaF and MgCl2. Of several leucocyte populations isolated from human blood by FACS, including T cells, B cells, NK lymphocytes and monocytes, ecto-PAPase was most prevalent on neutrophils; erythrocytes were essentially devoid of activity. A non hydrolysable, phosphonate analogue of phosphatidic acid, phosphonate 1, efficiently solubilized catalytic activity from intact neutrophils without causing cell disruption or increasing permeability. Enzyme activity in solubilized extracts was purified in the absence of detergent by successive heparin-Sepharose, gel filtration and anion exchange chromatography. By assaying activity in renatured SDS/polyacrylamide gel slices, the molecular mass of neutrophil ecto-PAPase was estimated to be between 45 and 52 kDa, similar to the molecular mass of previously purified plasma membrane PAPases. Since a large portion of neutrophil plasma membrane PAPase is available for hydrolysis of exogenous substrates, ecto-PAPase may play an important role in regulating inflammatory cell responses to extracellular phosphatidic acid in biological systems. PMID- 9210421 TI - Purification and properties of alpha-mannosidase II from Golgi-like membranes of baculovirus-infected Spodoptera frugiperda (IPLB-SF-21AE) cells. AB - An alpha-mannosidase II-like activity was identified in baculovirus-infected Spodoptera frugiperda (IPLB-SF21-AE) cells. The enzyme responsible was purified from Golgi-type membranes to apparent homogeneity by using a combination of steps including DEAE-cellulose, hydroxyapatite, concanavalin A-Sepharose and gel filtration chromatography. The molecular mass of this purified protein was approx. 120 kDa by SDS/PAGE under reducing conditions and approx. 240 kDa under non-reducing conditions, indicating that the enzyme is a disulphide-linked dimer. Substrates demonstrated to undergo hydrolysis with this enzyme were GlcNAc-Man5 GlcNAc-GlcNAc (non-reduced and reduced) and p-nitrophenyl alpha-d mannopyranoside. The oligosaccharide substrate was converted into GlcNAc-Man3 GlcNAc-GlcNAc through an intermediate GlcNAc-Man4-GlcNAc-GlcNAc. Treatment of the isolated intermediate oligosaccharide with endoglycosidase H resulted in its conversion into GlcNAc-Man4-GlcNAc. This indicated that it contained the alpha 1,3-linked mannose residue on the alpha-1,6-linked mannose arm and showed that the alpha-1,6-linked mannose residue on the alpha-1,6-linked mannose arm had been preferentially hydrolysed by the mannosidase. The oligosaccharide lacking the beta-1,2-linked GlcNAc residue on the alpha-1,3-linked mannose arm (Man5-GlcNAc GlcNAc) was not hydrolysed in the presence of the enzyme. Metal ions were not required for enzymic activity on any of the substrates, but Cu2+ was strongly inhibitory. The activity of the enzyme was inhibited at low concentrations of swainsonine, but much higher concentrations of 1-deoxymannojirimycin were required to achieve inhibition. All of these properties are characteristic of mannosidase II enzymes from other eukaryotic tissues. The presence of mannosidase II in lepidopteran insect cells would allow entry of N-linked glycoproteins into the complex processing reaction pathway or into the terminal Man3-GlcNAc-GlcNAc pathway. PMID- 9210422 TI - The binding-site sizes of Escherichia coli single-stranded-DNA-binding protein and mammalian replication protein A are 65 and >/= 54 nucleotides respectively. AB - The electrophoretic mobilities of complexes formed with single-stranded (ss) DNA and tetrameric Escherichia coli ssDNA-binding protein (EcoSSB) or mammalian replication protein A (RPA) were analysed. The electrophoretic mobilities of the complexes in a native polyacrylamide gel increased as the lengths of the DNA increased from 28 to 70 nt, thus revealing paradoxical 'descending-staircase' patterns. Increases in the electrophoretic mobilities of EcoSSB.ssDNA complexes were observed when the lengths of the bound DNA were increased by 1 nt. Quantitative analyses of the complexes suggested that the binding-sites sizes of EcoSSB and RPA were 65 and >=54 nt respectively. The binding-site size for RPA is at least 24 nt larger than previously reported. PMID- 9210423 TI - Type 1 ribosome-inactivating proteins are the most abundant proteins in iris (Iris hollandica var. Professor Blaauw) bulbs: characterization and molecular cloning. AB - The most abundant protein of Iris bulbs has been identified as a type 1 ribosome inactivating protein (RIP). Analysis of the purified proteins and molecular cloning of the corresponding cDNAs demonstrated that this type 1 RIP is a mixture of three isoforms that exhibit a high degree of sequence identity and have similar, though not identical, ribosome-inactivating and polynucleotide:adenosine glycosidase activities. The accumulation of large quantities of type 1 RIP in a vegetative storage organ suggests that this presumed defence-related protein also plays a role in the nitrogen-storage metabolism of the bulb. PMID- 9210425 TI - Increased flux through the hexosamine biosynthesis pathway inhibits glucose transport acutely by activation of protein kinase C. AB - The hexosamine biosynthesis pathway and protein kinase C (PKC) activation mediate hyperglycaemia-induced impaired glucose transport, but the relative role of each pathway is unknown. Following a 2 h preincubation of rat adipocytes in the presence of either high glucose (30 mM) plus insulin (0.7 nM) or glucosamine (3 mM), both high glucose and glucosamine inhibited subsequent basal and insulin stimulated glucose transport, measured at 5.0 mM glucose. Azaserine, an inhibitor of the enzyme glutamine:fructose-6-phosphate aminotransferase, abolished the effect of high glucose, but not that of glucosamine. Ro-31-8220, an inhibitor of PKC, reversed the effects of both high glucose and glucosamine, suggesting that flux through the hexosamine biosynthesis pathway impaired glucose transport acutely by activating PKC. Both high glucose and glucosamine caused a 3-fold increase in PKC activity; this effect of high glucose, but not that of glucosamine, was partially decreased by azaserine. Neither high glucose nor glucosamine altered basal or insulin-stimulated plasma membrane GLUT1 levels, whereas both treatments decreased basal, but not insulin-stimulated, GLUT4 levels. Azaserine abolished the effect of high glucose, but not that of glucosamine, on basal plasma membrane GLUT4 levels. Ro-31-8220, which returned glucose transport to control values, caused a further decrease in plasma membrane GLUT4 levels. It is concluded that, in rat adipocytes, an acute increase in flux through the hexosamine biosynthesis pathway inhibits glucose transport by activation of PKC. PMID- 9210424 TI - Oligomycin inhibits store-operated channels by a mechanism independent of its effects on mitochondrial ATP. AB - Inhibitors of mitochondrial oxidative metabolism have been proposed to interfere with Ca2+ influx mediated by store-operated channels (SOC), secondary to their effects on ATP production. We assessed SOC activity by 45Ca2+ influx and fluorimetric measurements of free Ca2+ or Mn2+ quench in thapsigargin-treated Chinese hamster ovary cells and Jurkat T-cells, and additionally by electrophysiological measurements of the Ca2+-release-activated Ca2+ current (Icrac) in Jurkat T-cells. Various mitochondrial antagonists were confirmed to inhibit SOC. However, the following evidence supported the proposal that oligomycin, in particular, exerts an inhibitory effect on SOC in addition to its known actions on mitochondria and Na+-pump activity: (i) the concentrations of oligomycin required to inhibit SOC-mediated Ca2+ influx or Icrac (half-inhibitory concentration approximately 2 microM) were nearly 50-fold higher than the concentrations that blocked mitochondrial ATP production; (ii) the rank order of potency of oligomycins A, B and C for decreasing SOC-mediated Ca2+ influx or Icrac differed from that known for inhibition of mitochondrial function; (iii) oligomycin blocked Icrac under voltage clamp and with intracellular Na+ and K+ concentrations fixed by dialysis from the patch pipette, arguing that the effect was not secondary to membrane polarization or pump activity; and (iv) fixing the cytosolic ATP concentration by dialysis from the patch pipette attenuated rotenone- but not oligomycin-mediated inhibition of Icrac. Oligomycin also blocked volume-activated Cl- currents, a profile common to some other known blockers of SOC that are not known mitochondrial inhibitors. These findings raise the possibility that oligomycin interacts directly with SOC, and thus may extend the known pharmacological profile for this type of Ca2+-influx pathway. PMID- 9210426 TI - A surface-plasmon-resonance analysis of polylysine interactions with a peptide substrate of protein kinase CK2 and with the enzyme. AB - The mechanism of protein kinase CK2 (CK2) activity stimulation by polylysine has been studied by surface plasmon resonance (SPR). The kinetics of the polylysine interaction with a peptide substrate of the enzyme, and with the enzyme itself, have been investigated. A peptide containing a threonine (T) residue surrounded by a cluster of negatively charged acidic [arginine (R) and glutamic acid (E)] residues, RRREEETEEE, and specifically phosphorylated by CK2, was selected. Polylysine interacts with both the enzyme and the peptide substrate. The rate constant, the stoichiometry of the polylysine-peptide substrate interaction and the kinetic parameters of the stimulated enzyme were used to calculate the polylysine-dependent stimulation of CK2. The results are in agreement with experimentally determined polylysine-dependent stimulation. The polylysine-enzyme interaction is too slow to account for enzyme stimulation. The behaviour of polylysine is not reproduced by the polyamine spermine. The results are consistent with a substrate-mediated mechanism of CK2 stimulation by polylysine, and they suggest that the CK2 stimulation by polyamines occurs by a different mechanism. PMID- 9210427 TI - Evaluation of extracellular lipid peroxidation in brain cortex of anaesthetized rats by microdialysis perfusion and high-performance liquid chromatography with fluorimetric detection. AB - A method for in vivo evaluation of lipid peroxidation in the extracellular space of anaesthetized rat brain cortex was developed. This method involved the use of microdialysis perfusion and high-performance liquid chromatography. The microdialysates, eluted from implanted probes, were reacted with thiobarbituric acid (TBA) prior to analysis by an HPLC system equipped with a fluorescence detector (excitation and emission wavelengths were 515 and 550 nm, respectively). Lipid peroxidation in the extracellular space was evaluated as the concentration of malondialdehyde, a lipid peroxidation end product which reacts with TBA to form a fluorescent conjugate. Significantly increased production of malondialdehyde following hydrogen peroxide perfusion (0.03%, 0.3% at a flow-rate of 1 microl/min) was observed in the brain cortex of anaesthetized rats. PMID- 9210428 TI - Determination of unchanged [18F]dopamine in human and non-human primate plasma during positron emission tomography studies: a new solid-phase extraction method comparable to radio-thin-layer chromatography analysis. AB - Routine determination of [18F]DOPA and its metabolites in plasma is essential for assessment and quantification of presynaptic dopamine function in vivo using a modeling approach with positron emission tomography (PET). The determination of unchanged [18F]DOPA from human and non-human primate plasma using solid-phase extraction (SPE) with Sep-Pak cartridges during PET dopaminergic studies is described here. The results from the studies showed that this new approach in comparsion to a method such as thin-layer chromatography (TLC) possessed a simplicity, rapidity and accuracy as well as good correlation between the two techniques (p<0.0001). A proposed procedure involving radioanalysis on alumina plates (Al2O3) was also developed with an excellent correlation compared to the conventional C18 plates (r=0.96). Thus it could be concluded that the SPE on either C18 or alumina cartridges (Waters) compared to radio-TLC analysis on C18 and alumina systems, appears to be a useful analytical method suitable for correcting the input arterial function in routine clinical PET neurotransmission studies. PMID- 9210429 TI - New sorbent for bilirubin removal from human plasma: Cibacron Blue F3GA immobilized poly(EGDMA-HEMA) microbeads. AB - Cibacron Blue F3GA-immobilized poly(EGDMA-HEMA) microbeads were investigated as a specific sorbent for bilirubin removal from human plasma. The poly(EGDMA-HEMA) microbeads were prepared by a modified suspension copolymerization technique. Cibacron Blue F3GA was covalently coupled to the poly(EGDMA-HEMA) microbeads via the nucleophilic reaction between the chloride of its triazine ring and the hydroxyl groups of the HEMA molecule, under alkaline conditions. Bilirubin adsorption was investigated from hyperbilirubinemic human plasma on the poly(EGDMA-HEMA) microbeads containing different amounts of immobilized Cibacron Blue F3GA, (between 5.0-16.5 micromol/g). The non-specific bilirubin adsorption on the unmodified poly(EGDMA-HEMA) microbeads were 0.32 mg/g from human plasma. Higher bilirubin adsorption values, up to 14.8 mg/g, were obtained with the Cibacron Blue F3GA-immobilized microbeads. Bilirubin molecules interacted with these sorbents directly. Contribution of albumin adsorption on the bilirubin adsorption was pronounced. Bilirubin adsorption increased with increasing temperature. PMID- 9210430 TI - Salt-independent adsorption of human serum proteins on cyanocarbon gels. AB - Electron donor acceptor gels based on cyanocarbons have been tested for human serum protein adsorption in the absence of salt-promotion by water-structuring salt. This phenomenon was compared with a normal adsorption process in the presence of salt. The tricyanoaminopropene-divinyl sulfone-agarose displayed unusual protein adsorption properties as binding could occur both independently or dependently of the salt-promotion. The absence of hydrophobic or ionic character of the salt-independent interaction suggests an electron donor acceptor adsorption mechanism which is shown, for the first time, to occur independently of salt-promotion in aqueous solution. Study of the protein adsorption specificity showed similar protein selectivity for the fractions adsorbed in both conditions. PMID- 9210431 TI - Determination of CTP synthetase activity in crude cell homogenates by a fast and sensitive non-radiochemical assay using anion-exchange high-performance liquid chromatography. AB - A non-radiochemical assay procedure for CTP synthetase was developed in which CTP is detected at 280 nm after separation with anion-exchange HPLC. A complete separation of all nucleoside triphosphates was achieved within 11 min and the minimum amount of CTP which could be accurately determined proved to be 5 pmol. Therefore, our assay procedure is ten-fold more sensitive compared to the frequently used radiochemical assays. The assay was linear with time and protein concentration, although at low protein concentration a lag phase was observed. An amount of 2 x 10(6) cells was already sufficient to determine the specific activity of CTP synthetase in HL-60 cells, lymphocytes and in lymphoblasts obtained from pediatric patients suffering from acute lymphoblastic leukemia. PMID- 9210432 TI - Determination of acetaldehyde in biological samples by gas chromatography with electron-capture detection. AB - A simple specific assay was developed for the determination of acetaldehyde in biological samples. Acetaldehyde was derivatized to 2,4-dinitrophenylhydrazone, which was determined by gas chromatography with electron-capture detection. The use of this detection method is an important device to which no one drew notice. This procedure was very simple and so sensitive that as little as 500 fmol of acetaldehyde could be measured in aqueous solution. The calibration curve of acetaldehyde was linear at least up to 40 microM. Its recoveries from human plasma and rat liver homogenate were 96.5 and 95.7%, respectively. PMID- 9210433 TI - Determination of cocaine and its metabolites in serum microsamples by high performance liquid chromatography and its application to pharmacokinetics in rats. AB - A single-solvent extraction step high-performance liquid chromatographic method is described for quantitating cocaine and its three metabolites in rat serum microsamples (50 microl). The separation used a 2.1-mm I.D. reversed-phase Brownlee C18 column with an isocratic mobile phase consisting of methanol acetonitrile-25.8 mM sodium acetate buffer, pH 2.2, containing 1.29-10(-4) M tetrabutylammonium phosphate (12.5:10:77.5, v/v/v). The detection limit was 2.5 ng/ml for all the compounds using an ultraviolet detector operated at 235 nm. The method was used to study the pharmacokinetics of cocaine after an intravenous (i.v.) bolus dose (4 mg/kg). PMID- 9210434 TI - Determination of LSD in blood by capillary electrophoresis with laser-induced fluorescence detection. AB - Capillary electrophoresis (CE) with HeCd laser-induced fluorescence (LIF) detection and its application in forensic toxicology is demonstrated by the determination of D-lysergic acid diethylamide (LSD) in blood. Following precipitation of proteins, washing of the evaporated supernatant and extraction, the residue was reconstituted in methanol and injected electrokinetically (10 s, 10 kV). The total analysis time for quantification of LSD was 8 min using a citrate-methanol buffer, pH 4.0. With this buffer system it is possible to separate LSD, nor-LSD, iso-LSD and iso-nor-LSD. Using a specific sample preparation, electrokinetic injection, extended light path (bubble cell) capillaries and especially LIF detection (lambda(ex) 325 nm, lambda(em) 435 nm), a limit of detection of 0.1-0.2 ng LSD per ml blood could be obtained. The limit of quantitation was about 0.4-0.5 ng/ml. The quantitative evaluation for LSD was carried out using methylergometrine as internal standard. The precision expressed as coefficient of variation (C.V.) and accuracy of the method were <20% and 86 110%, respectively. The application of the method to human blood samples from two forensic cases and a comparison with radioimmunoassay demonstrated that the results were consistent. PMID- 9210435 TI - Determination of residues of the beta-agonist clenbuterol in liver of medicated farm animals by gas chromatography-mass spectrometry using diphasic dialysis as an extraction procedure. AB - A method has been developed for the rapid confirmation of clenbuterol in cow liver using gas chromatography coupled with detection by mass spectrometry of the trimethylsilyl derivatives of clenbuterol. The technique used for the extraction was diphasic dialysis. It was observed that the best suitable solution to homogenize the liver the barium hydroxide-barium chloride buffer, the optimal extraction solvent was tert.-butylmethyl ether at an extraction temperature of 37 degrees C, and stirring should be applied at 150 rpm for 4 h. This extraction method improves clenbuterol recovery up to values of 99.3%. With the use of the barium buffer, derivatization is performed more efficiently and the detection and quantification limits can be decreased to values close to 250 ppt and 500 ppt, respectively. PMID- 9210436 TI - Measurement of dexfenfluramine metabolism in rat liver microsomes by gas chromatography-mass spectrometry. AB - A specific and useful method was developed for the determination of dexfenfluramine metabolism by microsomal systems utilising GC-MS. The synthesis of two metabolites 1-(3-trifluoromethylphenyl)propan-2-ol ('alcohol') and 1-(3 trifluoromethylphenyl)-1,2-propanediol ('diol') via straightforward routes, were confirmed by MS and NMR spectra. The conditions for extraction from alkalinised microsomal mixtures of the metabolites nordexfenfluramine, 1-(3 trifluoromethylphenyl)propan-2-one ('ketone'), alcohol and diol, their conversion to trifluoroacetate derivatives and analysis by GC-MS-SIM are described. Calibration curves were constructed between 48 and 9662 nM and fitted to quadratic equations (r2>0.999). The method precision was good over low (121 nM) medium (2415 nM) and above medium (9662 nM) concentrations for all metabolites; the within- and day-to-day coefficients of variation ranged between 2.5-12.4% and 6.7-17.5%, respectively. The accuracy, measured as bias, was very good both within- and day-to-day (range: -0.4-12.6%, 0.8-18.9%). For most metabolites, the C.V. for the assay and bias increased at 121 nM. Dexfenfluramine metabolism by rat liver microsomes was investigated using the assay method and showed a concentration dependent increase in nordexfenfluramine and ketone metabolites over the substrate range of 5-200 microM. PMID- 9210437 TI - Improvement of chemical analysis of antibiotics. XXIII. Identification of residual tetracyclines in bovine tissues by electrospray high-performance liquid chromatography-tandem mass spectrometry. AB - To reliably identify the residual tetracycline antibiotics (TCs), oxytetracycline (OTC), tetracycline, chlortetracycline (CTC) and doxycycline (DC), in bovine tissues, we have established a confirmation method using electrospray ionization liquid chromatography-tandem mass spectrometry (ESI LC-MS-MS) with daughter ion scan. All TCs gave [M+H-NH3]+ and [M+H-NH3-H2O]+ as the product ions, except for DC when [M+H]+ was selected as the precursor ion. The combination of C18 cartridge clean-up and the present ESI LC-MS-MS method can reliably identify TCs fortified at a concentration of 0.1 ppm in bovine tissues, including liver, kidney and muscle, and has been successfully applied to the identification of residual OTC in bovine liver and residual CTC in bovine muscle samples previously found at concentrations of 0.58 ppm and 0.38 ppm by LC, respectively. PMID- 9210438 TI - Analysis of terfenadine in human plasma using microbore high-performace liquid chromatography-electrospray ionisation mass spectrometry. AB - An assay based on combined microbore high-performance liquid chromatography positive ion electrospray ionisation mass spectrometry with selected ion recording has been developed for the measurement of the antihistamine drug terfenadine in human plasma. A deuterated analogue of terfenadine was synthesised for use as an internal standard and extraction of terfenadine was carried out on C18 solid phase extraction columns. The limit of detection of terfenadine in plasma is 0.1 ng/ml and the intra-assay coefficient of variation at 1 ng/ml is 10.1%. Plasma concentrations of terfenadine measured in six normal subjects following a 120 mg oral dose are reported. PMID- 9210439 TI - Direct determination of polyglucose metabolites in plasma using anion-exchange chromatography with pulsed amperometric detection. AB - High-performance anion-exchange chromatography with pulsed amperometric detection (HPAE-PAD) was evaluated for the quantitation of polyglucose metabolites (DP2 DP7) in human plasma. The method was investigated for accuracy, precision, specificity, linearity, range and analyte stability. Samples were prepared by dilution into the standard range (0.1-10 microg/ml) followed by deproteinization using a 30,000 molecular mass cut-off filtration device. The limit of detection was 0.05 microg/ml for all metabolites. Method precision for DP2-DP7 varied from approximately 2% R.S.D. in the upper range to approximately 15% R.S.D. at the limit of quantitation. Samples were stable following one or two freeze-thaw cycles and, after preparation, they could be refrigerated for up to 72 h. Application of this method to clinical plasma samples from continuous ambulatory peritoneal dialysis (CAPD) patients administered one daily night-time intraperitoneal exchange of 2 l of 7.5% polyglucose solution for four weeks indicated that plasma levels of DP2, DP3 and DP4 increased from baseline levels of <0.01 g/l to steady-state levels of 1.2+/-0.3, 1.2+/-0.3 and 0.4+/-0.1 g/l (mean+/-S.D.), respectively. These steady state plasma levels for DP2 and DP3 are comparable to previously reported levels in patients administered daily overnight 7.5% polyglucose dialysis solution. PMID- 9210440 TI - Isocratic high-performance liquid chromatographic method for the separation of isradipine and its main metabolites. Application to in vitro metabolization by h3A4/OR cells. AB - A reversed-phase HPLC method was developed for the study of isradipine oxidation in vitro. The drug and its main metabolites were determined after extraction from culture media and from h3A4/OR cells having the cytochrome P450 isoenzyme involved in the xenobiotic metabolization. The HPLC assay was fully validated and was used to follow the biotransformation kinetics of isradipine. PMID- 9210441 TI - Semi-preparative chromatographic purification of the enantiomers S-(-)-amlodipine and R-(+)-amlodipine. AB - Pharmacokinetic studies of optically pure compounds after single enantiomer administration are becoming increasingly important. The process of racemization in vivo can diminish all expected advantages of single enantiomer treatment. Amlodipine, one of the calcium channel blockers, currently used in therapy as a racemate, is one of such drugs under study. In order to administer single enantiomers of amlodipine to healthy volunteers both were chromatographically purified and characterised. The two optical isomers of amlodipine, active S-(-)- and non-active R-(+)-amlodipine, were purified using chromatographic procedure adopted from the analytical separation. Enantiomers were successfully converted to benzenesulphonic salt without any racemization. All semi-preparative purifications were monitored with complementary analytical methods, HPLC and CE, along with the determination of optical activity so that the final product was sufficiently defined for further in vivo studies. The analytical method developed for the determination of plasma concentrations of each enantiomer of amlodipine in these studies is also briefly described. PMID- 9210442 TI - Simultaneous determination of verapamil and norverapamil in biological samples by high-performance liquid chromatography using ultraviolet detection. AB - In this paper we develop an high-performance liquid chromatographic method with ultraviolet detection for the determination of verapamil and its primary metabolite norverapamil in biological samples. Both compounds, as well as the internal standard, imipramine, were extracted from alkalinised blood, with n hexane-isobutyl alcohol, back-extracted into 0.01 M phosphoric acid and determined using a reversed-phase column and ultraviolet monitoring at 210 nm. The average coefficient of variation obtained over the concentration range of 1 1000 ng/ml is about 3%. The detection limit is below 5 ng/ml for both compounds, and extraction recoveries close to 80%. The method was applied to a pharmacokinetic study of the drug and its active metabolite and used to analyse blood samples from verapamil treated rabbits. PMID- 9210443 TI - Determination and pharmacokinetics of a furosemide-amiloride drug combination. AB - The study presents an accurate and precise HPLC assay for the determination of furosemide and amiloride in human specimens. Both drugs were extracted from human plasma with ethyl acetate; furosemide was extracted at pH 1 and amiloride at pH 12. While chromatographic separation conditions, i.e., column, mobile phase and flow-rate were the same for both investigated drugs, furosemide was detected using a UV absorbance detector, whereas amiloride, because of its very low therapeutic range, was detected with a spectrofluorimetric detector. The linearity of the furosemide and amiloride assays were confirmed over the range of 30-3000 ng/ml and 0.5-30 ng/ml, respectively. These concentrations correspond well with the therapeutic ranges of both drugs. The extraction recoveries, depending on concentration, exceed 80% for furosemide and 74% for amiloride. The reported methods were applied to pharmacokinetic investigations of the two compounds taken in form of a drug combination. PMID- 9210444 TI - Liquid chromatographic method for the determination of ticlopidine in human plasma. AB - A simple high-performance liquid chromatographic method for determination of ticlopidine in human plasma using ultra violet detection was developed. The separation of the investigated compound and internal standard was achieved on a C18 BD column with a 0.01 M potassium dihydrogen phosphate buffer (pH 4) acetonitrile-methanol (20:40:40, v/v) mobile phase. The detection was performed at 215 nm. The compounds were isolated from plasma by Bond Elut C18 solid-phase extraction, the mean absolute recovery was 84.9%. The limit of quantitation was 10 ng ml(-1), the limit of detection was 5 ng ml(-1). The bioanalytical method was validated with respect to linearity, within- and between-day accuracy and precision. system suitability and stability. All validated parameters were found to be within the internationally required limits. The developed analytical method for ticlopidine was found to be suitable for application in pharmacokinetic studies and human drug monitoring. PMID- 9210445 TI - High-resolution liquid chromatographic method using ultraviolet detection for determination of ondansetron in human plasma. AB - This paper describes a simple technique for extraction and a sensitive high performance liquid chromatographic method for separation and quantitation of ondansetron in human plasma. The procedure involved liquid-liquid extraction of ondansetron from plasma, reversed-phase HPLC separation and ultraviolet detection at 305 nm. The internal standard method was applied for quantitation. The recovery of ondansetron was >85%. Linearity was good throughout the concentration range anticipated in human plasma from investigations in panic disorder (0.5-15 ng/ml, r2 ranging from 0.9953 to 0.9988). This method was applied to the determination of plasma concentrations of ondansetron in humans. PMID- 9210446 TI - Simultaneous rapid high-performance liquid chromatographic determination of phenytoin and its prodrug, fosphenytoin in human plasma and ultrafiltrate. AB - A reversed-phase high-performance liquid chromatographic assay for the simultaneous determination of phenytoin and fosphenytoin, a prodrug for phenytoin, in human plasma and plasma ultrafiltrate is described. For plasma, the method involves simple extraction of drugs with diethyl ether and evaporation of solvent, followed by injection of the reconstituted sample onto a reversed-phase C18 column. Plasma ultrafiltrate is injected directly into the HPLC column. Compounds are eluted using an ion-pair mobile phase containing 20% acetonitrile. The eluent is monitored by UV absorbance at 210 nm. The fosphenytoin standard curves are linear in the concentration range 0.4 to 400 microg/ml for plasma and 0.03 to 80 microg/ml for ultrafiltrate. Phenytoin standard curves are linear from 0.08 to 40 microg/ml for plasma and from 0.02 to 5.0 microg/ml for ultrafiltrate. No interferences with the assay procedure were found in drug-free blank plasma or plasma ultrafiltrate. Relative standard deviation for replicate plasma or ultrafiltrate samples was less than 5% at concentrations above the limit of quantitation for both within- and between-run calculations. PMID- 9210447 TI - Sensitive high-performance liquid chromatographic assay for aminoglycosides in biological matrices enables the direct estimation of bacterial drug uptake. AB - Following the development of a sensitive high-performance liquid chromatographic (HPLC) assay for gentamicin in biological matrices, the utility of this assay for the determination of other clinically important aminoglycosides (neomycin, netilmicin and sisomicin) in bacterial culture media or plasma is demonstrated. The high sensitivity of the assay enables direct measurement of the aminoglycoside content of bacterial cells cultured in the presence of unlabelled drug. PMID- 9210448 TI - High-performance liquid chromatographic determination of quinine in rat biological fluids. AB - A high-performance liquid chromatographic (HPLC) method with ultraviolet detection for the determination of quinine in rat biological fluids is described. Due to its selectivity and sensitivity, the proposed method can be used in the case of such rat biological fluids as cerebrospinal fluid (CSF) and perilymph for which the accessible volumes are limited to 100 microl and 10 microl, respectively. Consequently, the assay method has been applied to the measurements of quinine concentration in rat plasma, CSF and perilymph samples. PMID- 9210449 TI - Sensitive reversed-phase high-performance liquid chromatographic method for the determination of atevirdine and its N-desethyl metabolite in human saliva or cerebrospinal fluid using solid-phase extraction. AB - A sensitive reversed-phase high-performance liquid chromatographic method for the determination of atevirdine and its primary metabolite in human saliva or cerebrospinal fluid using solid-phase extraction is described. Samples mixed with internal standard and sodium phosphate buffer were applied to an activated C18 solid-phase extraction column. The reconstituted eluate was injected onto a Zorbax RX C8 column utilizing a mobile phase of 100 mM ammonium acetate (pH 4.0) isopropyl alcohol-acetonitrile (55:20:25, v/v/v). Fluorescence detection was employed with excitation at 295 nm and emission at 456 nm. Quantitation was achieved using peak-height ratios. The detection response curve was linear from 2 to 850 nM for atevirdine in both human saliva and cerebrospinal fluid and from 2 to 250 nM for the metabolite in human saliva. The method was utilized to analyze cerebrospinal fluid and saliva samples from clinical studies. PMID- 9210450 TI - Sensitive determination of docetaxel in human plasma by liquid-liquid extraction and reversed-phase high-performance liquid chromatography. AB - A sensitive reversed-phase high-performance liquid chromatographic method has been developed and validated for the quantitative determination of docetaxel (I) in human plasma. The concentrations in plasma, for validation procedures spiked with known amounts of I, are read from calibration curves in the range of 10 20,000 ng/ml. The sample preparation involved a liquid-liquid extraction of 1000 microl of sample with a mixture of acetonitrile-n-butylchloride (1:4, v/v). The related compound paclitaxel (II) was used as internal standard. Chromatographic separations were performed an Inertsil ODS-80A column, with UV detection performed at 230 nm. The overall extraction recoveries were 84.3 and 90.0% for I and II, respectively. The lower limit of quantitation was 10 ng/ml, and the accuracy, within-run and between-run precisions at three tested concentrations fell within the generally accepted criteria for bioanalytical assays. PMID- 9210451 TI - Use of post-column fluorescence derivatization to develop a liquid chromatographic assay for ranitidine and its metabolites in biological fluids. AB - Ranitidine and its main metabolites, ranitidine N-oxide and ranitidine S-oxide, were determined in plasma and urine after separation using reversed-phase liquid chromatography. The mobile phase consisted of an initial isocratic step with 7:93 (v/v) acetonitrile-7.5 mM phosphate buffer (pH 6) for 8 min, followed by a linear gradient up to a 25:75 (v/v) mixture over 1 min. Detection was carried out by a post-column fluorimetric derivatization based on the reaction of the drugs with sodium hypochlorite, giving rise to primary amines that reacted with o phthalaldehyde and 2-mercaptoethanol to form highly fluorescent products. The calibration graphs, based on peak area, were linear in the range 0.1-4 microg/ml for all drugs. The detection limits were 30, 41 and 32 ng/ml (8.6, 12.5 and 9.1 pmol) for ranitidine S-oxide, ranitidine N-oxide and ranitidine, respectively. Chromatographic profiles obtained for plasma and urine samples showed no interference from endogenous compounds. PMID- 9210452 TI - High-performance capillary electrophoresis measurement of dolastatin-10. AB - A high-performance capillary electrophoresis (HPCE) assay was used to determine the concentration of a potent cytotoxic agent, dolastatin-10, in human plasma. Following extraction from plasma, using a solid-phase C18 cartridge, capillary zone electrophoresis was used to separate, detect and quantitate dolastatin-10 using the structurally related compound dolastatin-15 as the internal standard. Migration times for both dolastatins are less than 20 min. The recovery of the drug was approximately 90% and was quantified over the assay range of 39 to 5000 ng/ml with good precision and accuracy. The method is linear up to 5000 ng/ml with a lower limit of detection of 25 ng/ml. Data resulting from the use of the assay for the in vitro metabolism of the drug are presented. This is the first report of a validated HPCE assay for determining dolastatin-10 levels in human plasma. PMID- 9210453 TI - Analysis of creatinine, vanilmandelic acid, homovanillic acid and uric acid in urine by micellar electrokinetic chromatography. AB - A simultaneous determination of vanilmandelic acid, homovanillic acid, creatinine and uric acid using capillary electrophoresis was investigated. The optimum conditions of buffer concentration, pH and surfactant concentration were studied, and high resolution was obtained using a 30 mM phosphate buffer (pH 7.0) containing 150 mM sodium dodecyl sulfate. The detection was by UV absorbance at 245 nm and the column was a fused-silica capillary of 67 cm x 75 microm I.D.. The determination of these metabolites in human urine was completed within 15 min without any interferences. PMID- 9210454 TI - Application of a sequential analytical procedure for the detection of the beta agonist brombuterol in bovine urine samples. AB - The multistep analytical procedure routinely applied in our laboratory for the detection of the aryl amine beta-agonists clenbuterol, mabuterol and mapenterol in bovine matrices has been extended to the analysis in urine samples of brombuterol, a new clenbuterol-like compound. In the screening steps, the urine samples were first enzyme-linked immunosorbent assay (ELISA)-tested, then the positive samples were analyzed by thin-layer chromatography (TLC) and the beta agonists detected with the Bratton-Marshall color reaction. The TLC spots corresponding to the suspected compounds were scraped off the plates, collected and extracted separately with methanol. The beta-agonists in the extracts were detected by HPLC-Vis (limits of detection: 0.5 ng/g). In the confirmatory step the presence and the concentration of the compounds of interest in the samples were established by GC-MS with two different ionization techniques, EI and CI (limits of detection: 1.0 ng/g). The use of this procedure has made possible the detection of brombuterol in officially sampled bovine urine. PMID- 9210455 TI - Rapid and sensitive high-performance liquid chromatographic assay for 6 hydroxychlorzoxazone and chlorzoxazone in liver microsomes. AB - A high-performance liquid chromatographic assay was developed for the quantitation of chlorzoxazone and its major metabolite 6-hydroxychlorzoxazone. These compounds along with phenacetin, the internal standard, were extracted from incubation mixtures using ether extraction. The extracts were analyzed on a Brownlee Spheri-5 C8 column with a mobile-phase of acetonitrile-0.5% phosphoric acid (30:70, v/v). The assay utilized UV detection at 287 nm which provided sensitivity and specificity to simultaneously quantify chlorzoxazone and 6 hydroxychlorzoxazone from liver microsomal samples at amounts of 10 ng and greater. The mean correlation coefficient of the standard curves for 6 hydroxychlorzoxazone and chlorzoxazone was 0.998 and 0.993, respectively, over the range of 25-400 ng, and the regression curves were found to be linear at least through 1600 ng. All components eluted within 7 min, resulting in a total analysis time of 8 min. The inter-day and intra-day coefficients of variation were <7 and <3%, respectively. This method provides a rapid, sensitive and cost effective assay for 6-hydroxychlorzoxazone and chlorzoxazone in liver microsomal incubations. PMID- 9210457 TI - Rapid and sensitive determination of amprolium in chicken plasma by high performance liquid chromatography with post-column reaction. AB - A rapid, sensitive and reproducible reversed-phase HPLC assay was developed for the determination of amprolium (APL) in chicken plasma. Protein in plasma sample was precipitated with 0.33 M perchloric acid and supernatant solution was injected into the HPLC system. Following the chromatographic separation of APL and the beclotiamine (I.S.) on a C18 column, the derivatives of APL and I.S. were formed by post-column reaction and detected by fluorescence detection (excitation at 400 nm, emission at 460 nm). The method showed excellent precision, accuracy and speed with a detection limit of 2 ng/ml. The intra- and inter-assay variance of this method were less than 11.2%. This method has been successfully applied to plasma determinations after oral administration of APL to chicken. PMID- 9210456 TI - Determination of dopaminergic prodrugs by high-performance liquid chromatography followed by post-column ion-pair extraction. AB - One possibility to optimize the therapeutic application of dopaminergic compounds with a catechol function is the reversible protection of this moiety using a prodrug approach. Important features in this respect are a proper chemical stability in the gastrointestinal tract, an adequate release rate after arrival in the blood stream or the possibility to cross the blood-brain barrier. A HPLC method was developed to measure the hydrolysis of prodrugs of dopamine and epinine directly. The method is based on reversed-phase separation followed by post-column ion-pair extraction with a fluorescent counter-ion. The separation of di-isobutyryl esters of dopamine and epinine is obtained within 10 min while the more hydrophobic dopaminergic esters, di-benzoyl and di-pivaloyl dopamine, are retained for 30 min. The precision of the assay measuring 160 ng dibudop and 100 ng ibopamine was 1.2 and 1.0%, respectively. The detection limit of all prodrugs tested was approximately 10 ng. PMID- 9210458 TI - Determination of the anticancer agent CI-980 in plasma by achiral liquid chromatography on a Pirkle-type stationary phase. AB - A high-performance liquid chromatographic assay has been developed and validated for the determination of the anticancer agent CI-980 in plasma samples from pediatric patients. After ether extraction from alkaline plasma, the CI-980 was chromatographed on a (R)-N-(3,5-dinitrobenzoyl)phenylglycine stationary phase using a mobile phase composed of hexane-isopropanol (70:30, v/v) modified with 1% acetonitrile. The method was linear over a 0.25 to 25.00 ng/ml range and the intra- and inter-day coefficients of variation (C.V.s) were less than 15%. The method was applied to the determination of the plasma concentration-time profile for a pediatric patient receiving 3.5 mg/m2 of CI-980 per day as a continuous 72 h infusion. PMID- 9210459 TI - Simple approach for analysis of plasma oxo-, hydroxy- and dicarboxylic acids. AB - Following deproteinization of plasma with organic solvents the supernatant was shaken with hexane and cation-exchange resin in an Eppendorf tube to remove fatty and amino acids and the medium was subjected to direct treatment with ethyl chloroformate under catalytic influence of pyridine. A subsequent extraction of the immediately formed ethyl esters with a drop of chloroform enabled us to subject the sample to gas chromatographic (GC) analysis. Since ketocarboxylic acids do not require a preliminary oximation the total time of sample workup and analysis takes only several minutes. PMID- 9210460 TI - Recognition and cleavage of DNA by type-II restriction endonucleases. AB - Restriction endonucleases are enzymes which recognize short DNA sequences and cleave the DNA in both strands. Depending on the enzymological properties different types are distinguished. Type II restriction endonucleases are homodimers which recognize short palindromic sequences 4-8 bp in length and, in the presence of Mg2+, cleave the DNA within or next to the recognition site. They are capable of non-specific binding to DNA and make use of linear diffusion to locate their target site. Binding and recognition of the specific site involves contacts to the bases of the recognition sequence and the phosphodiester backbone over approximately 10-12 bp. In general, recognition is highly redundant which explains the extreme specificity of these enzymes. Specific binding is accompanied by conformational changes over both the protein and the DNA. This mutual induced fit leads to the activation of the catalytic centers. The precise mechanism of cleavage has not yet been established for any restriction endonuclease. Currently two models are discussed: the substrate-assisted catalysis mechanism and the two-metal-ion mechanism. Structural similarities identified between EcoRI, EcoRV, BamHI, PvuII and Cfr10I suggest that many type II restriction endonucleases are not only functionally but also evolutionarily related. PMID- 9210461 TI - Cellular processing limits the heterologous expression of secretory component in mammalian cells. AB - Recombinant vaccinia-virus-based expression systems are very popular for the overproduction of proteins in mammalian cell lines. Both the double virus T7/vaccinia hybrid system and the single recombinant strategy based on the p11 K late promoter were evaluated for their ability to govern expression and secretion of recombinant human secretory component (SC), a glycoprotein associated with IgA in mucosal secretions. We report here that, while the T7 promoter is transcriptionally 3.4-fold more active than the p11 K promoter, no difference in levels of secreted recombinant human SC is observed using either vaccinia system to infect CV-1 cells. High transcription, and thus translation levels, lead to saturation of early processing steps involved in protein export. Both systems exhibit transient accumulation of comparable amount of recombinant human SC in the endoplasmic reticulum and/or the cis Golgi network, as demonstrated by immunofluorescence and endoglycosidase H (EndoH) sensitivities. Exposure of infected cells to tunicamycin results in similar inhibition of recombinant human SC export, further arguing that N-linked glycosylation is necessary for proper folding and subsequent secretion. Moreover, pulse-chase experiments indicate that newly synthesized recombinant human SC is not completely processed in a mature glycoprotein and that a portion of overexpressed SC might be degraded before it can be secreted. Recombinant human SC behaves identically to native SC in terms of kinetics of secretion and IgA-binding capacity. Our results indicate that optimization of expression systems should not only rely on the design of effective vectors, but also on the identification and clearance of the cellular bottlenecks associated with maturation of the secreted proteins. PMID- 9210462 TI - Molecular localisation of ferrochelatase in higher plant chloroplasts. AB - Within the chloroplast of higher plants, a crucial branchpoint of the tetrapyrrole synthesis pathway is the chelation of either Fe2+ to make haem, or Mg2+ for chlorophyll, catalysed by ferrochelatase or magnesium chelatase, respectively. One model that has been proposed for the control of this branchpoint, based on biochemical studies, is that the two enzymes are spatially separated within the chloroplast, ferrochelatase being exclusively in the thylakoids, while magnesium chelatase is associated with the envelope [Matringe, M., Camadro, J.-M., Joyard, J. & Douce, R. (1994) J. Biol. Chem. 269, 15010 15015]. We have used a sensitive molecular method to investigate this possibility. Radiolabelled precursor proteins for ferrochelatase from Arabidopsis have been imported into isolated chloroplasts. Their distribution in the different subchloroplastic fractions have then been determined, and compared with that for light-harvesting chlorophyll protein, which is exclusively thylakoidal, and the envelope-located phosphate translocator. Clear evidence for the specific association of ferrochelatase protein with both thylakoid and envelope membranes has been obtained, thus suggesting strongly that the control of the branchpoint cannot be by spatial separation of the two chelatases. PMID- 9210463 TI - The N-terminal Arg-rich region of human immunodeficiency virus types 1 and 2 and simian immunodeficiency virus Nef is involved in RNA binding. AB - Comparison of the amino acid sequences of human immunodeficiency virus (HIV) Nef protein and several RNA-binding proteins shows similarities in some regions of these proteins. Thus, poliovirus protein 2C, an RNA-binding protein, shares with Nef the sequence YXQQ...MDD...DXXD. In addition, both proteins contain an Arg rich motif that, in the case of poliovirus 2C, is involved in RNA-binding activity. Moreover, the RNA-binding, anti-terminator N proteins of lambda, phi21 and P22 phages show sequence similarities with HIV Nef at the Arg-rich motif. To assess the significance of this motif, native and deletion variants of Nef protein were assayed for RNA-binding activity. The N-terminal 35 amino acids of HIV-1 Nef that comprise the Arg-rich motif are sufficient for RNA binding. Point mutations engineered at the Arg-rich motif of HIV-1 Nef revealed that basic amino acid residues are essential for RNA-binding activity. The Nef proteins from HIV-2 and SIV can also interact with RNA, while the same proteins with the N-terminal Arg-rich domain truncated fail to interact with RNA. These findings indicate that all three Nef proteins from HIV-1, HIV-2 and simian immunodeficiency virus belong to the RNA-binding family of proteins. The three proteins contain an Arg-rich region at the N-terminus which is necessary to interact with RNA. PMID- 9210464 TI - Influence of the Gloeophyllum metabolite oosponol and some synthetic analogues on protein and RNA synthesis in target cells. AB - Oosponol [4-(hydroxyacetyl)-8-hydroxy-1H-2-benzopyran-1-one], a toxic metabolite of the basidiomycete Gloeophyllum abietinum, and some synthetic analogues were studied to clarify the molecular basis of their fungicidal, bactericidal, and phytotoxic effects. The antibiotic activity was due to a strong inhibition of RNA and protein synthesis in target cells. PMID- 9210465 TI - Cloning of an annelid fibrillar-collagen gene and phylogenetic analysis of vertebrate and invertebrate collagens. AB - Arenicola marina possesses cuticular and interstitial collagens, which are mostly synthesised by its epidermis. A cDNA library was constructed from the body wall. This annelid cDNA library was screened with a sea-urchin-collagen cDNA probe, and several overlapping clones were isolated. Nucleotide sequencing of these clones revealed an open reading frame of 2052 nucleotides. The translation product exhibits a triple helical domain of 138 Gly-Xaa-Yaa repeats followed by a 269 residue-long C-terminal non-collagenous domain (C-propeptide). The triple helical domain exhibits an imperfection that has been previously described in a peptide produced by cyanogen bromide digestion (CNBr peptide) of A. marina interstitial collagen. This imperfection occurs at the same place in the interstitial collagen of the vestimentiferan Riftia pachyptila. This identifies the clone as coding for the C-terminal part of a fibrillar collagen chain. It was called FAm1alpha, for fibrillar collagen 1alpha chain of A. marina. The non-collagenous domain possesses a structure similar to carboxy-terminal propeptides of fibrillar pro alpha chains. Only six conserved cysteine residues are observed in A. marina compared with seven or eight in all other known C-propeptides. This provides information on the importance of disulfide bonds in C-propeptide interactions and in the collagen-assembly process. Phylogenetic studies indicate that the fibrillar collagen 1alpha chain of A. marina is homologous to the R. pachyptila interstitial collagen and that the FAm1alpha gene evolved independently from the other alpha-chain genes. Complementary analyses indicate that the vertebrate fibrillar collagen family is composed of two monophyletic subgroups with a specific position of the collagen type-V chains. PMID- 9210466 TI - A conformational study of the human and rat encephalitogenic myelin oligodendrocyte glycoprotein peptides 35-55. AB - Myelin oligodendrocyte glycoprotein (MOG), is considered an important central nervous system-specific target autoantigen for primary demyelination in autoimmune diseases like multiple sclerosis. We have recently demonstrated that MOG or its derived peptide, MOG-(35-55)-peptide, are able to produce in animals, clinicopathologic signs that mimic multiple sclerosis. The rat MOG sequence spanning amino acids 35-55 [rMOG-(35-55)-peptide] differs from the human sequence [hMOG-(35-55)-peptide] by a single amino acid substitution, i.e. Pro42-->Ser. Mice injected with rMOG-(35-55)-peptide showed severe inflammation and demyelination throughout the central nervous system but, interestingly, mice injected with hMOG-(35 -55)-peptide showed only a few foci of mild inflammation with no demyelination. Circular dichroism and nuclear magnetic resonance spectroscopy have been used to structurally characterise the bioactive peptides hMOG-(35-55)-peptide and rMOG-(35-55)-peptide. In 0.1 M K2HPO4/KOH, 90% H2O/D2O solutions, these derived peptides have been shown, by NMR spectroscopy, to adopt detectable levels of short-range structure in equilibrium with unfolded conformers. On addition of 2,2,2-trifluoro-(2H3)ethanol, rMOG-(35-55)-peptide and hMOG-(35-55)-peptide adopt folded structures which have nuclear Overhauser enhancements characteristic of a poorly defined alpha-helix over residues 44-51. There are some indications of secondary structure also evident in the N-terminal region of rMOG-(35-55)-peptide. CD spectroscopy has revealed that in aqueous solution both peptides are unfolded but in 2.2.2-trifluoroethanol and, at micellar concentrations of sodium dodecyl sulfate, rMOG-(35-55)-peptide and, to a lesser extent, hMOG-(35-55)-peptide adopt helical conformations. In contrast, at non-micellar concentrations of SDS rMOG-(35-55)-peptide and hMOG-(35-55)-peptide adopt, according to CD spectroscopy, a beta-structure indicating that the peptides change conformation depending on the microenvironment of the amino acids. PMID- 9210467 TI - Characterization and purification from bovine neutrophils of a soluble guanine nucleotide-binding protein that mediates isozyme-specific stimulation of phospholipase C beta2. AB - Members of the beta isozyme subfamily of phosphatidylinositol-specific phospholipase C (PLC) are stimulated by alpha subunits and betagamma dimers of heterotrimeric guanine-nucleotide-binding proteins (G proteins). Myeloid differentiated human HL-60 granulocytes and bovine neutrophils contain a soluble phospholipase C, which is stimulated by the metabolically stable GTP analogue guanosine (5'-->O)-3-thiotriphosphate (GTP[S]). To identify the component(s) involved in mediating this stimulation, the relevant polypeptide(s) was resolved from endogenous phospholipase C and purified from bovine neutrophil cytosol by measuring its ability to confer GTP[S] stimulation to exogenous recombinant PLCbeta2. The resolved factor, which behaved as 48-kDa protein upon gel filtration, stimulated PLCbeta2 but not PLCbeta1 or PLCdelta1. Activation of phosphatidylinositol 4-phosphate 5-kinase was not involved in this stimulation. The purified stimulatory factor consisted of two polypeptides of molecular masses of approximately 23 kDa and 26 kDa. The protein stimulated a deletion mutant of PLCbeta2 that lacked a carboxyl-terminal region necessary for stimulation by members of the alpha(q) subfamily of the G-protein alpha subunits. The results of this study suggest that a GTP-binding protein distinct from alpha(q) subunits, probably a low-molecular-mass GTP-binding protein associated with a regulatory protein, is involved in isozyme-specific activation of PLCbeta2. PMID- 9210468 TI - Memory and imprinting effects in multienzyme complexes--I. Isolation, dissociation, and reassociation of a phosphoribulokinase-glyceraldehyde-3 phosphate dehydrogenase complex from Chlamydomonas reinhardtii chloroplasts. AB - A bienzyme complex made up of phosphoribulokinase and glyceraldehyde-3-phosphate dehydrogenase has been isolated and purified from chloroplasts of Chlamydomonas reinhardtii. The complex contains four phosphoribulokinase and eight glyceraldehyde-3-phosphate dehydrogenase polypeptide chains. As phosphoribulokinase is dimeric and glyceraldehyde-3-phosphate dehydrogenase tetrameric, it is concluded that the complex comprises two phosphoribulokinase and two glyceraldehyde-3-phosphate dehydrogenase molecules. Its overall molecular mass is 460 kDa, which is in excellent agreement with its stoichiometry. Moreover, owing to the nature of the two enzymes, this complex must catalyse two nonconsecutive reactions. The bienzyme complex tended to spontaneously dissociate into the free enzymes upon dilution. This dissociation process was considerably promoted by reducing agents such as dithiothreitol or reduced thioredoxin. The kinetics of the dissociation process induced by dithiothreitol or reduced thioredoxin were paralleled by an increase of activity of phosphoribulokinase. The dissociation of the complex was reversible. If oxidized phosphoribulokinase and glyceraldehyde-3-phosphate dehydrogenase were mixed, a certain amount of the complex was formed. The reconstituted complex displayed properties that were indistinguishable from those of the native complex extracted from chloroplasts of Chlamydomonas reinhardtii. These results suggest that the concentration of the complex in vivo must vary depending on the light intensity. PMID- 9210469 TI - Memory and imprinting effects in multienzyme complexes--II. Kinetics of the bienzyme complex from Chlamydomonas reinhardtii and hysteretic activation of chloroplast oxidized phosphoribulokinase. AB - Oxidized, free, stable phosphoribulokinase from Chlamydomonas reinhardtii was almost completely devoid of catalytic activity (0.06 s(-1)/site). However, when it was bound to glyceraldehyde-3-phosphate dehydrogenase from the same organism, it displayed significant activity (3.25 s(-1)/site). Moreover, this complex tended to spontaneously dissociate upon dilution; the isolated phosphoribulokinase activity increased up to 56 s(-1)/site, subsequently decreased, and finally became almost completely inactive. Its intrinsic kinetic properties (Km and k(cat)) changed with the variation of the overall activity. These effects were paralleled by changes of conformation of the enzyme as revealed by fluorescence analysis. A model is proposed that allows quantitative expression of the dynamics of the dissociation of the oxidized bienzyme complex and the effects of either of the two substrates, ATP and ribulose 5-phosphate, on this dissociation process. Whereas ATP destabilized the complex and promoted its dissociation, ribulose 5-phosphate tended to stabilize this complex. Inactive, stable, oxidized phosphoribulokinase may form a complex with glyceraldehyde-3 phosphate dehydrogenase regaining its catalytic activity. In this case, glyceraldehyde-3-phosphate dehydrogenase acts in a manner similar, but not identical to a chaperonin. The information content of the phosphoribulokinase gene, as defined by the sequence of its base pairs, was therefore not sufficient to specify full enzyme activity. It needed the presence of glyceraldehyde-3 phosphate dehydrogenase to give the oxidized phosphoribulokinase a conformation competent for its activity. The potential biological significance of these effects remains to be discovered. PMID- 9210470 TI - Comparison of the uptake and processing of cholesterol from chylomicrons of different fatty acid composition in rats fed high-fat and low-fat diets. AB - The fate of [3H]cholesterol carried in chylomicrons prepared from rats given a meal of palm oil (rich in long-chain saturated fatty acids), olive oil (rich in monounsaturated fatty acids) or corn oil (rich in n-6 polyunsaturated fatty acids) was investigated in vivo in rats fed a low-fat diet or a diet supplemented with the corresponding oil (to provide 40% of the calories) for 21 days. In the low-fat-fed groups, radioactivity was removed from the blood and secreted into bile over 180 min more rapidly when the chylomicrons were derived from corn oil as compared to palm or olive oil. After feeding the corresponding high-fat diets, however, both parameters were decreased in rats fed palm and corn oil, but not olive oil. As a result of these changes, the rates of removal of radioactivity from the blood and secretion into bile were similar in animals given the olive oil and corn oil diets, and higher than those in rats fed the palm oil diet. All the high-fat diets tended to increase the proportion of the radioactivity in the plasma found in the 1.006-1.050-g/ml fraction (low-density lipoprotein) and decrease that in the 1.050-1.25-g/ml (high-density lipoprotein) fraction in comparison to the respective low-fat diet groups, but the transfer of radioactivity to the plasma high-density lipoprotein fraction was particularly slow in palm-oil-fed rats. These findings indicate that diets high in saturated or n-6 polyunsaturated fat retard the metabolism of chylomicron cholesterol in comparison to diets low in fat, while those high in monounsaturated fat do not have this effect. As a consequence of this, the rate of removal of cholesterol of dietary origin from the body is slower in animals fed saturated as compared to monounsaturated or n-6 polyunsaturated fat. Thus, differential metabolism of chylomicron cholesterol clearly plays an important role in the hyper- and hypo cholesterolaemic effects of these dietary fats. PMID- 9210471 TI - The nuclear ABC1 gene is essential for the correct conformation and functioning of the cytochrome bc1 complex and the neighbouring complexes II and IV in the mitochondrial respiratory chain. AB - The nuclear ABC1 gene was isolated as a multicopy suppressor of a cytochrome b mRNA translation defect. Its inactivation leads to a respiratory deficiency suggesting a block in the bc1 segment of the respiratory chain [Bousquet, I., Dujardin, G. & Slonimski, P. P. (1991) EMBO J. 10, 2023-2031]. In the present study, we established that deleting the ABC1 chromosomal gene from Saccharomyces cerevisiae does not prevent the assembly of the bc1 complex (complex III) but markedly impairs the kinetics of its high-potential electron transfer pathway occurring on the positive, outer, side of the membrane, which results in reduced activity of the bc1 complex. In addition, the activity of complex II and its cytochrome b560 decrease drastically and complex IV activity is halved. It is also observed that the binding of the quinol to the bc1 complex ubiquinol oxidation site is affected and that adding exogenous quinones partially compensates for the respiratory deficiency in vitro, although the quinone content of mutant and wild-type mitochondria are similar. Lastly, complexes II, III and IV are found to be thermosensitive and the bc1 complex exhibits greater sensitivity than the wild-type strain to center N and P inhibitors, suggesting that the three multisubunit complexes have undergone structural modifications. The data suggest that the ABC1 gene product acts as a chaperone-like protein essential for the proper conformation and efficient functioning of the bc1 complex and the effects of the Abc1 protein on the complexes II and IV might result from interactions with the modified bc1 complex. PMID- 9210473 TI - Characterisation of the molybdenum-responsive ModE regulatory protein and its binding to the promoter region of the modABCD (molybdenum transport) operon of Escherichia coli. AB - Molybdenum-dependent repression of transcription of the Escherichia coli modABCD operon, which encodes the high-affinity molybdate transporter, is mediated by the ModE protein. This regulatory protein was purified as an N-terminal His6-tagged derivative and characterised both with and without the N-terminal oligohistidine extension. Equilibrium centrifugation showed that ModE is at least a 57-kDa homodimer. Circular dichroism spectroscopy indicated that when molybdate or tungstate bind to ModE there is little change in its alpha-helical content, but a major change in the environment of tryptophan and tyrosine residues occurs. Addition of molybdate or tungstate to the protein results in almost 50% quenching of the fluorescence attributed to tryptophan. Titration of fluorescence quenching showed that two molecules of molybdenum bind to each dimer of ModE with a Kd of 0.8 microM. DNA mobility-shift assays showed that ModE requires molybdenum, or tungstate, to bind with high affinity (approximate Kd of 30 nM ModE) to the modABCD promoter region. In accord with ModE's role as a molybdenum-dependent transcriptional repressor, DNase I footprinting experiments showed that the ModE molybdenum complex binds to a single 31-bp region around the transcription start of the modABCD promoter. This region contains a 6-base palindromic sequence CGTTAT-N12-ATAACG. PMID- 9210472 TI - Dexamethasone increases expression of 5-lipoxygenase and its activating protein in human monocytes and THP-1 cells. AB - The aim of this study was to assess the effect of dexamethasone on 5-lipoxygenase pathway expression in human peripheral blood monocytes and the acute monocytic leukemia cell line, THP-1. Cells were conditioned over a period of days with dexamethasone, at concentrations relevant in vivo, to study the effect of the glucocorticoid on calcium-ionophore-stimulated 5-lipoxygenase product and arachidonic acid release. The effect of dexamethasone on levels of immunoreactive protein and steady-state messenger RNA encoding for 5-lipoxygenase and its activating protein (5-LAP) was also assessed. Dexamethasone increased the stimulated release of 5-lipoxygenase products from both monocytes and THP-1 cells in a dose-dependent fashion. The increase in product generation was not due to changes in the availability of arachidonic acid. However, immunoreactive protein and steady-state messenger RNA encoding for 5-lipoxygenase and 5-LAP were increased by conditioning with dexamethasone. There was no apparent effect of the glucocorticoid on LTA4-hydrolase-immunoreactive protein levels or specific activity. We conclude that dexamethasone increases 5-lipoxygenase pathway expression in both monocytes and in THP-1 cells. This effect is due, at least in part, to increases in immunoreactive protein and steady-state messenger RNA encoding for 5-lipoxygenase and 5-LAP. These results suggest a role for glucocorticoids in the regulation of 5-lipoxygenase pathway expression in mononuclear phagocytes. PMID- 9210474 TI - Activation of partially folded mitochondrial malate dehydrogenase by thioredoxin. AB - CD spectroscopy reveals that mitochondrial malate dehydrogenase in 3M guanidinium chloride shows little residual secondary structure. Refolding of the denatured protein by dilution with buffer of pH 7.5 does not restore the CD spectrum of the native enzyme. A partially folded intermediate, possessing 25% of the alpha-helix content of the native enzyme, is formed upon dilution. The partially folded intermediate binds the extrinsic probe 1-anilinonaphtalene-8-sulfonate, and the increase in fluorescence (tenfold) is accompanied by a blue shift in the band position of the emission spectrum. Partially folded malate dehydrogenase is devoid of catalytic activity. In vitro refolding of the denatured protein takes place in the presence of dithiotreitol and thioredoxin. In the presence of micromolar concentrations of thioredoxin, a recovery of approximately 70% of the catalytic activity was observed. Emission-anisotropy titrations of oxidized thioredoxin, tagged with a fluorescent probe, revealed that the oxidoreductase recognizes partially folded intermediates of malate dehydrogenase with a dissociation constant of 6 microM. Moreover, a covalently linked complex formed by thioredoxin and monomeric malate dehydrogenase was detected by SDS/PAGE. A general mechanism is postulated for the reactivation of denatured proteins by thioredoxin. PMID- 9210475 TI - An aspartic endopeptidase is involved in the breakdown of propeptides of storage proteins in protein-storage vacuoles of plants. AB - To understand the mechanism of the maturation of various proteins in protein storage vacuoles, we purified a 48-kDa aspartic endopeptidase composed of 32-kDa and 16-kDa subunits from castor bean. Immunocytochemical and cell fractionation analyses of the endosperm of maturing castor bean seed showed that the aspartic endopeptidase was localized in the matrix of the protein-storage vacuoles, where a variety of seed storage proteins were also present. The amount of the aspartic endopeptidase increased at the mid-maturation stage of the seeds before accumulation of the storage proteins. To determine how the aspartic endopeptidase is responsible for maturation of seed proteins in concert with the vacuolar processing enzyme, we prepared 35S-labeled proproteins of seed proteins from the endoplasmic reticulum fraction of pulse-labeled maturing endosperm and used the authentic proproteins as substrates for in vitro processing experiments. The purified aspartic endopeptidase was unable to convert any of three endosperm proproteins, pro2S albumin, proglobulin, and proricin, into their mature sizes, while the purified vacuolar processing enzyme could convert all three proproteins. We further examined the activity of aspartic endopeptidase on the cleavage of an internal propeptide of Arabidopsis pro2S albumin, which is known to be removed post-translationally. The aspartic endopeptidase cleaved the propeptide at three sites under acidic conditions. These results suggest that aspartic endopeptidase cannot directly convert pro2S albumin into the mature form, but it may play a role in trimming the C-terminal propeptides from the subunits that are produced by the action of the vacuolar processing enzyme. PMID- 9210476 TI - Nuclear factor Y controls the basal transcription activity of the mouse platelet derived-growth-factor beta-receptor gene. AB - To determine the regulatory mechanism of the expression of the mouse platelet derived growth factor (PDGF) beta-receptor gene, a 1.9-kb 5' flanking genomic fragment was cloned and analyzed. Site-directed mutagenesis of a CCAAT motif, located 60 bp upstream of the transcriptional-start site, completely abolished the promoter activity [Ballagi, A. E., Ishisaki, A., Nelin, J.-O. & Funa, K. (1995) Biochem. Biophys. Res. Commun. 210, 165-1751. The sequence around the intact CCAAT motif was protected by in vitro DNase-I-footprinting analysis. Electrophoresis-mobility-shift assays with anti-[nuclear factor Y(NF-Y)]Ig revealed binding of the NF-Y complex to the CCAAT box. Furthermore, the double stranded oligonucleotides corresponding to the sequence around the CCAAT motif were conjugated with DNA-affinity magnetic beads. The binding proteins were affinity purified and identified as the NF-Y transcription factor by western blotting. Our results indicate that NF-Y controls the basal transcription activity of the mouse PDGF beta-receptor gene. PMID- 9210477 TI - Stimulation of Sky tyrosine phosphorylation by bovine protein S--domains involved in the receptor-ligand interaction. AB - Protein S is an anticoagulant vitamin-K-dependent plasma glycoprotein, which acts as a cofactor to activated protein C in the degradation of coagulation factors Va and VIIIa. It has been proposed that protein S has an additional function as a growth factor. Protein S and a structurally similar protein, Gas6, have been found to stimulate members of the Axl/Sky family of receptor tyrosine kinases. Human Gas6 is able to activate Axl and Sky. In contrast, while bovine protein S activates human Sky and its murine homologue, human protein S activates murine Sky but not the human receptor. In the present investigation, we studied the structural background of this species difference. Using protein S chimeras with domains from human and bovine origin, we found that only those chimeras with the steroid-hormone-binding globulin-like (SHBG) region from bovine protein S activate human Sky, indicating that the SHBG region is essential for the interaction. This observation was confirmed by inhibition of Sky phosphorylation by C4b-binding protein, a plasma protein that interacts tightly with the SHBG region of protein S. Another chimeric molecule, composed of the N-terminal 4 carboxyglutamic-acid-containing domain (Gla domain) and the two epidermal-growth factor-like domains of human factor IX, and the SHBG region of bovine protein S, stimulated the receptor less efficiently. Antibodies directed against the Gla domain of protein S, inhibited the activation of human Sky by bovine protein S. These results indicate that the N-terminal domains of protein S are not essential for activation of the receptor, but contribute to the affinity of the interaction. Our data suggest that protein S might be a ligand of Sky in some species despite the lack of activity of human protein S on human Sky. The bovine/human protein S species difference will be a useful model to establish the structural requirements for the interaction between Sky and its ligands. PMID- 9210478 TI - The mouse gene for hypoxia-inducible factor-1alpha--genomic organization, expression and characterization of an alternative first exon and 5' flanking sequence. AB - The ubiquitously expressed hypoxia-inducible factor-1 (HIF-1) is involved in expression of a large number of oxygen-regulated genes. HIF-1 is a heterodimer consisting of an alpha and a beta subunit, both belonging to the basic-helix-loop helix Per-aryl hydrocarbon receptor nuclear translocator-Sim (PAS) family of transcription factors. Whereas HIF-1alpha is a novel member of this family, HIF 1beta is identical to the aryl hydrocarbon receptor nuclear translocator, previously recognized to be involved in xenobiotic metabolism. cDNA cloning revealed that mouse HIF-1alpha can be expressed as two mRNA isoforms containing alternative 5' untranslated regions and two different predicted translational start sites. We cloned and characterized 20.5 kb of the mouse HIF-1alpha gene (Hif1a) containing exon II-XV. The two alternative first exons, I.1 and I.2, are separated from exon II by approximately 24 kb and 17 kb, respectively. We also sequenced Hif1a exon I.1 and flanking regions, and mapped a single exon I.1 transcription initiation site. Reverse transcription PCR analysis of total RNA derived from normoxic and hypoxic mouse hepatoma and fibroblast cell lines suggested that the two alternative mRNA isoforms are constitutively coexpressed in these cells, and that two different promoters drive transcription of HIF 1alpha. A minimal exon I.1 promoter was identified which moderately activated heterologous gene expression, indicating that additional cis-elements are required for efficient HIF-1alpha transcription in vivo. PMID- 9210479 TI - The N-terminal region of alpha-dystroglycan is an autonomous globular domain. AB - The structure of the N-terminal region of mouse alpha-dystroglycan (DGN) was investigated by expression of two protein fragments (residues 30-180 and 30-438) in Escherichia coli cells. Trypsin susceptibility experiments show the presence of a stable alpha-dystroglycan N-terminal region (approximately from residue 30 to 315). In addition, guanidinium hydrochloride (Gdn/HCl) denaturation of DGN-(30 438)-peptide, monitored by means of tryptophan fluorescence, produces a cooperative transition typical of folded protein structures. These results strongly suggest that the alpha-dystroglycan N-terminal is an autonomous folding unit preluding a flexible mucin-like region and that its folding is not influenced by the absence of glycosylation. In order to obtain more information on the structural features of the N-terminal domain we have also used circular dichroism, analytical sedimentation and electron microscopy analysis. Circular dichroic spectra show the absence of typical secondary structure (e.g. alpha helix or beta-sheet) and closely resemble those recorded for loop-containing proteins. This is consistent with a sequence similarity of the alpha-dystroglycan domain with the loop-containing protein elastase. Analytical ultracentrifugation and electron microscopy analysis reveal that the N-terminal domain has a globular structure. DGN-(30-438)-peptide does not bind in the nanomolar range to an iodinated agrin fragment which binds with high affinity to tissue purified alpha dystroglycan. No binding was detected also to laminin. This result suggests that the alpha-dystroglycan N-terminal domain does not contain the binding site to its extracellular matrix binding partners. It is less likely than the lack of glycosylation reduces its binding affinity, because the N-terminal globular domain only contains two glycosylation sites. PMID- 9210480 TI - Mutational analysis of extracellular cysteine residues of rat secretin receptor shows that disulfide bridges are essential for receptor function. AB - We attempted to express point-mutant secretin receptors where each of the 10 extracellular Cys residues was replaced by a Ser residue, in Chinese hamster ovary (CHO) cells. Six of the point-mutant receptors (C24-->S, C44-->S, C53-->S, C67-->S, C85-->S and C101-->S) could not be detected by binding or functional studies: the mutations resulted in functional inactivation of the receptor. In contrast, the four other point-mutant receptors (C11-->S, C186-->S, C193-->S and C263-->S) were able to bind poorly 125I-secretin, and to activate adenylate cyclase with high secretin EC50 values. These results suggest that cysteine residues 24, 44, 53, 67, 85 and 101 are necessary for receptor function, and that the two putative disulfide bridges formed by cysteine residues 11, 186, 193 and 263 are functionally relevant, but not essential for receptor expression. Secretin activated the adenylate cyclase through the quadruple mutant (C11,186,193,263-->S), the four triple mutants, and through double mutants C186,193-->S and C186,263-->S with a very high (microM) EC50 value, suggesting that, in the wild-type receptor, disulfide bridges are formed between C11-C186, and between C193-C263. Prior treatment with dithiothreitol resulted in a marked EC50 increase of the wild-type receptor and of those receptors with at least the two cysteine residues in positions 11 and 186, suggesting that the C11-C186 (but not the C193-C263) disulfide bridge was accessible to this reducing agent. Several results nevertheless indicated that, in mutant receptors, alternative disulfide bridges can be formed between cysteine 186 and cysteine 193 or 263, suggesting that these three residues are in close spatial proximity in the wild type receptor. PMID- 9210481 TI - The complete cDNA sequence and expression of the first major allergenic protein of Malassezia furfur, Mal f 1. AB - For the first time the complete cDNA encoding a major allergen and novel protein of the yeast Malassezia furfur, Mal f 1, has been sequenced and expressed. The amino acid sequences of nine tryptic peptides of the protein were determined. Oligonucleotides were designed from these amino acid sequences. The cDNA sequence was obtained by hybridizing these primers to mRNA and enhancement by reverse transcriptase PCR techniques. The cDNA is 1176 bp in length. It shows an open reading frame of 1050 bp coding for a protein of 38178 Da and a deduced amino acid sequence containing 350 residues. The hydropathy plot and the tryptic digest indicate that the first 22 amino acids represent a leader sequence determining a mature protein of 35 988 Da. The complete encoding cDNA was expressed as a maltose-binding protein fusion protein in Escherichia coli. The recombinant fusion protein reacted with our specific monoclonal antibody and with IgE from patients with atopic dermatitis. PMID- 9210482 TI - Up-regulation of P-glycoprotein expression in rat liver cells by acute doxorubicin treatment. AB - Expression of P-glycoprotein, a plasma-membrane glycoprotein involved in multidrug resistance and encoded by mdr genes, was investigated in cultured rat liver cells acutely exposed to doxorubicin. This anticancer drug was shown to increase mdr mRNA levels in a dose-dependent manner in both rat liver epithelial (RLE) cells and primary rat hepatocytes. This induction of mdr transcripts was detected as early as a 4-h exposure to doxorubicin used at 0.5 microg/ml. It occurred through increased expression of the mdr1 gene as assessed by northern blot analysis using rat mdr-gene-specific probes. In addition, RLE cells exposed to doxorubicin displayed an overexpression of a 140-kDa P-glycoprotein as demonstrated by western blotting. Moreover, doxorubicin-treated RLE cells displayed enhanced cellular efflux of the P-glycoprotein substrate rhodamine 123 that was inhibited by the P-glycoprotein blocker verapamil, thus providing evidence that doxorubicin-induced P-glycoprotein was functional in liver cells. Doxorubicin-mediated mdr mRNA induction was found to be fully inhibited by actinomycin D, thus indicating its dependence on RNA synthesis; it was demonstrated to be not associated with alteration of protein synthesis, suggesting it differed from the known mdr mRNA overexpression occurring in response to cycloheximide. In contrast to P-glycoprotein, other liver detoxification pathways such as cytochromes P-450 1A were not induced by doxorubicin treatment. These data indicate that doxorubicin can act as a potent acute inducer of functional P-glycoprotein in rat liver cells and therefore may modulate both chemosensitivity of hepatic cells and P-glycoprotein-mediated biliary secretion of xenobiotics. PMID- 9210483 TI - The lantibiotic mersacidin inhibits peptidoglycan biosynthesis at the level of transglycosylation. AB - The lantibiotic mersacidin has been previously reported to interfere with bacterial peptidoglycan biosynthesis, [Brotz, H., Bierbaum, G., Markus, A., Molitor, E. & Sahl, H.-G. (1995) Antimicrob. Agents Chemother. 39, 714-719]. Here, we focus on the target reaction and describe a mersacidin-induced accumulation of UDP-N-acetylmuramoyl-pentapeptide, indicating that inhibition of peptidoglycan synthesis occurs after the formation of cytoplasmic precursors. In vitro studies involving a wall-membrane particulate fraction of Bacillus megaterium KM demonstrated that mersacidin did not prevent the synthesis of lipid II [undecaprenyl-diphosphoryl-N-acetylmuramoyl-(pentapeptide)-N-ac ety lglucosamine] but specifically the subsequent conversion of this intermediate into polymeric nascent glycan strands by transglycosylation. Comparison with other inhibitors of transglycosylation shows that the effective concentration of mersacidin in vitro is in the range of that of the glycopeptide antibiotic vancomycin but 2-3 orders of magnitude higher than that of the competitive enzyme inhibitor moenomycin. The analogy to the glycopeptides may hint at an interaction of mersacidin with the peptidoglycan precursor rather than with the enzyme. Unlike vancomycin however, mersacidin inhibits peptidoglycan formation from UDP-N acetylmuramoyl-tripeptide and is active against Enterococcus faecium expressing the vanA resistance gene cluster. This indicates that the molecular target site of mersacidin differs from that of vancomycin and that no cross-resistance exists between the two antibiotics. PMID- 9210485 TI - Influence of Zn2+ on the kinetic events that contribute to the 500-kDa dextran sulfate-dependent activation of factor XII (Hageman factor). AB - The effect of zinc ions on kinetic and equilibrium steps that may contribute to the activation and subsequent autoactivation of human blood coagulation factor XII in the presence of 500-kDa dextran sulfate was studied. To analyze the results, the expression of the overall autoactivation constant that had been established from the model presented in a previous study was used. Comparison of the kinetics obtained at different levels of zinc, which included amounts lower than the residual concentration introduced by NaCl in the incubation mixture, suggested that the addition of Zn2+ up to 5 microM lowered the mean number of sites available for the binding of factor XII to the surface from 220 to 172 and increased the rate of the first-order activation of factor XII by one order of magnitude, from (1.6 +/- 0.4) x 10(-4) s(-1) to (8.0 +/- 0.4) x 10(-4) s(-1) in the presence of 550 nM dextran sulfate. Neither the factor XII/surface dissociation constant (1 microM), the apparent catalytic constant, nor the apparent Michaelis-Menten constant associated with the postulated multi-stage kinetic sequence were affected by the presence of zinc. Most experimental trends induced by the presence of zinc could be successfully interpreted by using the model, thus reinforcing its reliability under different conditions. PMID- 9210484 TI - Characterisation of the pterin molybdenum cofactor in dimethylsulfoxide reductase of Rhodobacter capsulatus. AB - Analysis of dimethylsulfoxide reductase from Rhodobacter capsulatus showed that it contained 1 mol Mo and 2 mol GMP. This indicates that the molybdenum cofactor in dimethylsulfoxide reductase is bis(molybdopterin guanine dinucleotide) molybdenum. The absorption spectrum of the molybdopterin guanine dinucleotide released from dimethylsulfoxide reductase after denaturation of the holoenzyme was compared with those of pterin standards of known redox state. The spectra were most similar to pterin standards in the dihydro state and oxidised state. The reduction of 2,6-dichloroindophenol by molybdopterin guanine dinucleotide released from dimethylsulfoxide reductase and by pterin standards was also measured and approximately 2 mol electrons/2 mol molybdopterin guanine dinucleotide were found to reduce 2,6-dichloroindophenol. These results are consistent with the presence of one molybdopterin guanine dinucleotide moiety with a pyrazine ring at the oxidation level of a dihydropteridine and one molybdopterin guanine dinucleotide moiety with a pyrazine ring at the oxidation level of a fully aromatic pteridine. It is suggested that the pyrazine ring of Q molybdopterin guanine dinucleotide is fully aromatic and contains a 5,6 double bond. PMID- 9210486 TI - Detection of metal binding to bovine inositol monophosphatase by changes in the near and far ultraviolet regions of the CD spectrum. AB - Mg2+ ions, essential for the catalytic activity of mammalian inositol monophosphatase, increase the ellipticity in the near-ultraviolet region of the CD spectrum of the enzyme. These spectral changes are not affected by the additional presence of substrate and are reversed if EDTA is added to the solution of enzyme and metal ions. Titration of the spectral perturbation at 275 nm shows that this binding occurs with a dissociation constant (Kd) around 275 microM, 292 microM and 302 microM for the wild-type, [Gln217]inositol monophosphatase and [Phe219]inositol monophosphatase enzymes respectively. The source of the spectroscopic change at 275 nm is not Trp219. The addition of Mg2+ also causes a decrease in ellipticity over most of the far-ultraviolet region of the spectrum (between 205-240 nm). The Kd values describing the binding of Mg2+ ions are 3.9 mM, 6.8 mM and 29.1 mM for the wild-type, [Gln217]inositol monophosphatase and [Phe219]inositol monophosphatase enzymes, respectively, each showing an approximate 12% change in ellipticity. In the additional presence of 10 mM Pi, there is a fourfold increase in the affinity of wild-type enzyme for Mg2+. It is concluded that CD spectral changes at wavelengths around 275 nm are indicative of metal ions interacting with a high-affinity metal-binding site (site 1). The spectral changes around 225 nm are associated with interactions at a lower-affinity site normally occupied by the Mg2+ ion which is reflected by the Km value for this metal ion. Other metal ions such as Ca2+ and Tb3+ (but not Mn2+ or Zn2+) also perturb the CD spectrum of the enzyme in both regions of the spectrum. The amplitudes of these signal changes are greater for Mg2+ or Tb3+ (25%) ions than for Ca2+ (8.5%), although two Ca2+-binding sites with Kd values of 20 microM and 100 microM have been identified. The uncompetitive inhibitor Li+ causes little change in the near-ultraviolet spectrum in the absence or presence of either substrate or Pi. However, in contrast to other metal ions, Li+ ions elicit a 10% increase in ellipticity at 220 nm with a Kd of 0.8 mM. PMID- 9210487 TI - Solution structure of Lqh-8/6, a toxin-like peptide from a scorpion venom- structural heterogeneity induced by proline cis/trans isomerization. AB - Lqh-8/6 is a minor fraction isolated from the venom of the scorpion Leiurus quinquestriatus hebraeus. Here we describe the purification, amino acid sequencing and solution structure determination by NMR and molecular modeling of this peptide. Lqh-8/6 is a small polypeptide (38 residues) which contains 8 half cystines and is highly similar to another venom component, chlorotoxin. Standard homonuclear methods were used to sequentially assign the proton NMR spectra and to collect spatial restraints for structure determination. Two populations, identified early in the assignment step, are in slow interconversion on the NMR timescale. The two conformers were shown to originate from a cis/trans peptidyl prolyl isomerization. Using a distance geometry program and simulated annealing protocol under the NMR restraints we obtained 10 final structures for the major conformation (trans isomer). None of the structures showed NOE violations larger than 0.05 nm, and the rmsd value relative to the mean structure (considering the main chain atoms in well-defined secondary structure) is 0.07 nm. The three dimensional structure contains a short alpha-helix strapped on a small antiparallel beta-strand and an N-terminal extended fragment. The sequence/structure and structure/function relationships of the new scorpion toxin like peptide are discussed in the context of the present structure determination. This toxin shows a stable, highly populated cis conformer of a peptidyl-prolyl peptide bond. PMID- 9210488 TI - Complex ATP-activation kinetics of plant H+-transporting ATPase may or may not require two substrate sites. AB - The complex ATP-activation kinetics of plant H+-ATPase requires two ATP effects on the enzyme. They may result from the simultaneous existence of two ATP sites or a single site that consecutively changes its properties. We describe here three main models for ATP binding to the plant H+-ATPase. Considering the experimental data there are some restrictions in their application. A system with two simultaneous catalytic sites with cooperative binding is possible provided the substrate exerts regulatory properties, and when ES (or SE) leads to a slower velocity path than SES (v1 < v2). In other words, simple cooperativity does not work. A system with two substrate sites, one of which is catalytic and the other is always and only regulatory (i.e. it affects the overall reaction rate), offers two alternatives: one with potential cooperative binding (the system does not discriminate between binding to the catalytic and regulatory sites; and the other with intrinsically different affinities of catalytic and regulatory sites (i.e. the system discriminates between binding to the two binding sites). Here it is also obligatory that ES (or SE) leads to a slower-velocity path than SES (v1 < v2). Thirdly, a system is possible with single ATP domain that is consecutively catalytic and regulatory as the cycle proceeds. These three mechanisms give rise to equivalent rate equations. Therefore, there is no way to distinguish between them on the basis of kinetic studies. Another conclusion drawn from modeling these schemes is that the form of the plots might resemble but not correspond to certain cooperativity type. For instance, for two substrate sites, a true negative cooperativity for substrate binding can mimic positive cooperativity if the v1 velocity pathway is much slower than the v2 one. PMID- 9210489 TI - HNK-1 carbohydrate-mediated cell adhesion to laminin-1 is different from heparin mediated and sulfatide-mediated cell adhesion. AB - The sulfated HNK-1 carbohydrate present on glycolipids and on several neural recognition molecules has been shown to mediate the adhesion of murine small cerebellar neurons and astrocytes to the extracellular matrix molecule laminin-1. In this study, we characterized the binding of the HNK-1 carbohydrate to laminin 1 extracted from the Engelbreth-Holm-Swarm (EHS) sarcoma and distinguished it unequivocally from binding sites for other sulfated carbohydrates. Electron microscopic analysis of rotary shadowed complexes of laminin-1 and a HNK-1 neoglycoprotein revealed a major binding site on the G domain that comprises the C-terminal globule of the laminin alpha1 chain. The HNK-1 carbohydrate also interacted with placental laminin isoforms containing an alpha chain variant. It bound to the proteolytic laminin-1 fragment E8 comprising the domains G1-G3, but not to fragment E3 that carries the major heparin-binding site on domains G4-G5. No binding was observed to the short arm containing fragments E1XNd or P1. Binding studies with native or denatured laminin E8 fragments and proteolytic or recombinant fragments of the G domain localized the HNK-1 carbohydrate binding site to domain G2. The binding could be clearly distinguished from binding sites for other sulfated carbohydrates such as heparin and sulfatides. Further, the binding could not be abolished by reduction and alkylation or by urea treatment of laminin-1 and was independent of the native conformation of laminin-1 and of Ca2+. The G2 domain is also involved in the adhesion of HNK-1 carbohydrate expressing early postnatal cerebellar neurons and is different from heparin- and sulfatide-mediated cell adhesion to laminin-1. HNK-1 carbohydrate-mediated cell adhesion appears, however, to be dependent on the native conformation of laminin 1 indicating a more complex cellular recognition process. PMID- 9210490 TI - Domain-specific N-glycosylation of the membrane glycoprotein dipeptidylpeptidase IV (CD26) influences its subcellular trafficking, biological stability, enzyme activity and protein folding. AB - Dipeptidyl peptidase IV (DPPIV, CD26) is an N-glycosylated type II plasma membrane protein. The primary structure of rat wild-type DPPIV contains eight potential N-glycosylation sites. To investigate the role of N-glycosylation in the function of DPPIV, three of its asparagine residues were separately converted to glutamine by site-directed mutagenesis. The resulting N-glycosylation mutants of rat DPPIV were studied in stable transfected Chinese hamster ovary cells. All three N-glycosylation mutants of DPPIV showed a reduced half-life, as well as differing degrees of inhibition of the processing of their N-glycans. Mutation of the first (Asn83-->Gln) or eighth (Asn686-->Gln) N-glycosylation site had only a small effect on its enzymatic activity, cell-surface expression and dimer formation, whereas the mutation of the sixth N-glycosylation site (Asn319-->Gln) abolished the enzymatic activity, eliminated cell-surface expression and prevented the dimerization of the DPPIV protein. The mutant [Gln319]DPPIV is retained in the cytoplasm and its degradation was drastically increased. Our data suggest that the N-glycosylation at Asn319 is involved in protein trafficking and correct protein folding. PMID- 9210492 TI - Factors involved in rejection of concordant xenografts in complement-deficient rats. AB - BACKGROUND: Factors that contribute to xenograft (Xg) rejection were investigated in complement C6-deficient (C-) PVG rats. METHODS: First and second hamster hearts were transplanted in C6-deficient and C6-sufficient PVG rats. Xenoantibody (XAb) formation, hemolytic C (CH50) activity and immunohistochemistry were studied. RESULTS: PVG C6-deficient rats rejected Xgs 3 days later than PVG C6 sufficient rats. Surprisingly, C activation participated in the rejection in PVG C- rats, as shown by partially recovered serum CH50 levels and deposition of C factors in the Xgs. As we found that cultured endothelial cells produced C6 in vitro, we hypothesized that Xg endothelial cells corrected the C6 defect in PVG C rats. This was probably induced by IgM XAbs as: (1) it did not occur in immunosuppressed PVG C- rats in which XAb formation was prevented, and (2) transfer of IgM XAbs to naive, xenotransplanted PVG C- rats accelerated the recovery of CH50 and concomitantly Xg rejection. Thirty days after rejection of a first Xg, when no IgM XAbs or CH50 activity but high levels of IgG XAbs were detected in PVG C- rats, second Xgs underwent a hyperacute rejection. This time, complement was not involved, as no serum CH50 nor C deposition was found in the Xg. Instead, IgG antibody-dependent cellular cytotoxicity was involved as: (1) IgG XAbs were deposited in the Xg and (2) hyperacute rejection was induced in naive PVG C- rats by transfer of IgG XAbs, and (3) this rejection was delayed to 5+/-3 days if the adoptive hosts were first irradiated. CONCLUSIONS: In the face of a defect of host C factors, IgM XAb may induce cells of the Xg to secrete C factors which may correct the C defect of the host. Even if activation of lytic C can be prevented, IgG XAb may still provoke an acute Xg rejection by antibody dependent cellular cytotoxicity. PMID- 9210491 TI - The glycosylated matrix protein of Borna disease virus is a tetrameric membrane bound viral component essential for infection. AB - Borna disease virus (BDV) is representative of the family of Bornaviridae in the order Mononegavirales (negative-stranded, non-segmented, enveloped RNA viruses). It is the causal agent for Borna disease, characterized as an encephalomyelitis (typical form) in a wide variety of domestic animals (from rodents to birds). Recent information shows the involvement of BDV in the pathogenesis of some human psychiatric disorders. The 8.9-kb viral antigenome codes for five major ORF. The third ORF codes for a 16-kDa protein (matrix protein) that is posttranslationally modified, yielding an N-linked glycoprotein. Our data show that the glycosylated matrix protein exists as a stable tetrameric structure detectable either by electrospray ionization or matrix-assisted laser-desorption ionization mass spectrometry. Under native conditions, the tetramer, with a relative molecular mass of 68 kDa, was isolated from a sediment-free brain suspension of a BDV infected horse. The 68-kDa entity is stable in the presence of ionic and nonionic detergents but dissociates into subunits when heated. We found that the tetrameric matrix protein inhibits in vitro BDV infection in a dose-dependent manner. In contrast to inhibition of BDV infection with hydrophobic carbohydrate derivatives and protein-bound glycoconjugates, the glycosylated matrix protein is a very potent inhibitor of BDV infection, indicating that this protein represents an essential virus-specific membrane component for viral attachment. PMID- 9210494 TI - Evidence for survival and metabolic activity of encapsulated xenogeneic hepatocytes transplanted without immunosuppression in Gunn rats. AB - BACKGROUND: Hepatocyte transplantation could be an alternative to whole organ transplantation to correct enzymatic disorders. To this end, it would be of major importance to use xenogeneic cells without immunosuppression. The aim of this study was to investigate the survival and metabolic activity of encapsulated xenogeneic hepatocytes in the absence of immunosuppression. For this purpose, we used Gunn rats genetically incapable of bilirubin conjugation. METHODS: Xenogeneic (from guinea pigs) and allogeneic (from Lewis rats) hepatocytes (2x10(7)) were isolated, macroencapsulated in hydrogel hollow fibers made with an acrylonitrile-sodium methallyl-sulfonate copolymer, and transplanted into the peritoneum of Gunn rats without any immunosuppression. Plasma bilirubin levels were evaluated weekly. Bilirubin conjugates in bile and cell morphology were studied after 5 and 12 weeks, respectively. RESULTS: In Gunn rats transplanted with xenogeneic hepatocytes, a significant decrease in the serum bilirubin level was observed between 3 and 9 weeks after transplantation when compared with controls transplanted with empty hollow fibers: it fell to 62% of the initial level at weeks 5-7 (P < 0.01). A comparable result was observed in Gunn rats transplanted with encapsulated allogeneic cells. Bilirubin conjugates were observed in bile samples of rats transplanted with encapsulated hepatocytes. After explantation, hollow fibers appeared intact with minimal fibrosis. Cell viability and hepatocyte morphology were preserved. CONCLUSIONS: These results indicate that macroencapsulated xenogeneic hepatocytes can survive and remain functional for more than 2 months when transplanted in vivo in the absence of any immunosuppression. PMID- 9210493 TI - Assessment of insulin secretion in vitro from microencapsulated fetal porcine islet-like cell clusters and rat, mouse, and human pancreatic islets. AB - BACKGROUND: The possibility of transplanting microencapsulated pancreatic islets into patients with insulin-dependent diabetes mellitus, either as allografts or xenografts, has attracted great interest. A critical evaluation of the results obtained reveals that the success has been very limited. The aim of the present study was to compare the in vitro function of microencapsulated islets obtained from adult humans, adult mice, adult rats, and fetal pigs. METHODS: Human pancreatic islets were isolated at beta-Cell Transplant in Brussels, Belgium, and sent to the Department of Medical Cell Biology, Uppsala University in Uppsala, Sweden. Rat and mouse pancreatic islets and fetal porcine islet-like cell clusters (ICC) were prepared in Uppsala. All groups of islets were subsequently sent to the Department of Biotechnology, Norwegian Institute of Biotechnology, University of Trondheim, Trondheim, Norway. After 1 day in tissue culture, the islets were microencapsulated in alginate then cultured and sent back to Uppsala the next day. After either overnight culture (day 1) or 6 days of culture (day 6), the microencapsulated islets were examined for their insulin content and insulin release. Nonencapsulated islets from the same isolations were used as controls. RESULTS: The insulin content of rodent and human islets was not affected by microencapsulation, whereas porcine ICC showed a diminished insulin content. Microencapsulated porcine ICC also had a marked reduction in their insulin secretion in response to stimulation with glucose or glucose + theophylline both on days 1 and 6 in tissue culture. Mouse islets showed a reduced insulin response at both time points. Rat islets exhibited an inhibition of insulin secretion on day 1, but this had been restored by day 6. Human islets had well-preserved insulin secretion after both days 1 and 6. Microencapsulated human islets showed a normal morphology 3-4 weeks after intraperitoneal transplantation to nude mice. CONCLUSIONS: Pancreatic islets isolated from human, rat, and mouse donors show a glucose-stimulated insulin release in vitro after microencapsulation and repeated transports between laboratories. The insulin secretory capacity of microencapsulated human and rat islets was preserved best, whereas mouse islets and particularly fetal porcine ICC were impaired by microencapsulation. PMID- 9210495 TI - Functional and morphological outcome of knee joint transplantation in dogs depends on control of rejection. AB - BACKGROUND: The reconstruction of massive osteochondral defects extending to weight-bearing joints remains a surgical challenge. Total knee joint transplantation has been performed experimentally, but these studies lacked prospective evaluation of functional outcome, graft vascularization, and graft viability. METHODS: Replantation and transplantation of vascularized knee joints was performed in dogs (n=4 per group), comparing functional and morphological results during a 6-month follow-up. RESULTS: All replant recipients and three transplant recipients survived the 6-month follow-up period. At this time, duplex sonography and angiography revealed patent anastomoses in all animals. Increases in volumetric flow rates and vascular collateralization were observed in allografts, as compared with replanted joints (100+/-16 ml/min vs. 31+/-15 ml/min at 6 months after transplantation). Bone fusion at the graft-host interface was verified by fluorography in all animals at 3 months after transplantation. Six months after transplantation, microradiographies and computerized tomographies revealed spongialization of the cortical bone and filling of the medullary space by trabecular bone in transplanted joints. Such alterations were not detectable in replanted joints. Chondrocyte viability exceeded 80% in all but one transplanted joint. Lymphocyte infiltration of synovia and arterial walls was detected in all transplanted joints, suggesting the presence of chronic rejection. Weight-bearing capacity recovered in all replanted animals (weight bearing index before transplantation: 0.499+/-0.080; 6 months after transplantation: 0.38+/-0.16) but only in two of four transplanted animals (weight-bearing index 6 months after transplantation: 0.37, 0.28, and 0.00). CONCLUSIONS: These data demonstrate the potential of joint grafting and the critical dependence of allotransplantation on the control of rejection. PMID- 9210496 TI - Use of the methylxanthine derivative A802715 in transplantation immunology: II. In vivo experiments. AB - BACKGROUND: We have previously demonstrated in vitro that the methylxanthine derivative A802715 suppresses the cyclosporine (CsA)-resistant "signal two" dependent pathway of T cell activation and hence acts synergistically with CsA. Here, this synergism was further investigated in vivo in rats. METHODS: Primary cardiac allografts were placed in the neck, and secondary grafts were transplanted intra-abdominally. A802715 was given orally for 30 days or by continuous intravenous infusion via a mini-osmotic pump for 2 weeks. CsA was given orally for up to 30 days. T cell responses were examined in vitro using mixed lymphocyte reaction, concanavalin A whole blood, and cell-mediated lympholysis assays. RESULTS: In a major histocompatibility complex incompatible WKAH-->PVG combination, neither oral CsA (7.5 mg/kg/day) nor oral A802715 (100 mg/kg/day) was able to prolong graft survival. However, a combination of both drugs, given at the same dose, sustained graft survival during treatment. A similar synergism was not obtained with pentoxifylline, another methylxanthine derivative. The synergism between A802715 and CsA could be further increased by using a continuous intravenous infusion of A802715, since (1) lower doses of A802715 (20 mg/kg/day) and CsA (5 mg/kg/day) could be used, and (2) six of seven grafts survived permanently. In a major histocompatibility complex compatible Wag/Rij-->R/A combination, similar synergistic effects and permanent graft survival could also be obtained by oral A802715 (100 mg/kg/day) in combination with a low dose of CsA (2.5 mg/kg/day). In both strain combinations, long-term survivors accepted donor-type but rejected third-party second grafts in the absence of immunosuppression. This specific tolerance was not related to clonal deletion nor anergy, as recipient lymphocytes proliferated normally in the anti donor mixed lymphocyte reaction. Instead, a defect in generating specific cytotoxic T lymphocytes was involved. CONCLUSIONS: A802715 synergizes with CsA in vivo to induce specific transplantation tolerance and hence should be considered as a promising new immunosuppressant. PMID- 9210497 TI - Absence of deleterious effect on long-term kidney graft survival of rejection episodes with complete functional recovery. AB - BACKGROUND: Rejection episodes (RE) exert a detrimental influence on long-term kidney graft outcome. However, the impact of the severity of those RE on graft survival and the factors that could predict this impact are ill defined. The present retrospective study was undertaken on adult patients who received 582 cadaver kidney transplants at our center during the last 12 years, to assess the impact on graft survival of RE occurring during the first year after transplantation and to uncover the factors associated with the severity of those RE. METHODS: Three grades of rejection were defined: (1) rejection without loss of graft function (benign rejection); (2) rejection followed by partial loss of graft function (severe rejection); and (3) rejection with return to dialysis (irreversible rejection). The grafts were distributed among four groups: (1) grafts free of rejection; (2) grafts with benign RE (only grade 1 RE); (3) grafts with severe RE (one or more grade 2 RE); and (4) grafts with irreversible (grade 3) RE. RESULTS: Multivariate analyses revealed that (1) the occurrence of RE during the first posttransplant year (group 1 versus groups 2, 3, and 4) was significantly associated with primary immunosuppression with CsA rather than with OKT3 monoclonal antibody, the number of HLA-B + DR mismatches, and the younger recipient's age; (2) in patients with rejection, OKT3 monoclonal antibody prophylaxis was less often used in patients with irreversible RE (group 4) than in those with reversible RE (group 2, benign, and group 3, severe); and (3) no single factor was able to differentiate patients with benign RE (group 2) from those with severe RE (group 3). For grafts still functioning 1 year after transplantation, long-term graft survival was similar in grafts with either no RE or benign RE, but it was significantly lower (P<0.0001) in grafts with severe RE: 8-year survival rates were 89% and 60%, respectively. The decline in graft survival after 1 year was significantly correlated with the serum creatinine value but not with the dose of cyclosporine at 1 year. CONCLUSIONS: Benign RE occurring during the first year after transplantation and resulting in no loss of graft function do not exert a detrimental influence on long-term kidney graft outcome. In contrast, the prognosis of grafts with severe RE during the same period of time is much poorer. PMID- 9210498 TI - Determination of HLA-A,B residue mismatch acceptability for kidneys transplanted into highly sensitized patients: a report of a collaborative study conducted during the 12th International Histocompatibility Workshop. AB - BACKGROUND: During the 12th International Histocompatibility Workshop, a collaborative study between 35 laboratories was conducted on a group of highly allosensitized patients who had received a kidney transplant from 1981 to 1995. The major goal of the study was to assess how serum screening against a large cell panel could determine donor HLA mismatch acceptability in relation to graft outcome. METHODS: Twenty laboratories participated in an extensive screening of 92 high panel-reactive antibody (PRA) sera from patients at 29 transplant centers worldwide; each patient had received a kidney allograft from an HLA-A,B mismatched unrelated donor. Screening was done by complement-dependent lymphocytotoxicity and antihuman globulin augmentation techniques using a common protocol and shared standardized reagents. After an extensive quality-control assessment, we selected data from 14 participants who had screened the sera against a combined panel of 535 HLA-typed cells. RESULTS: With the 2x2 table based Multiscreen computer program, we could readily determine for virtually every patient the significant correlations between serum reactivity and the presence of panel cell markers, including private and public HLA-A,B epitopes and amino acid residues assigned from published sequencing data. Donor mismatch acceptability was assessed at the amino acid residue level. In the complement dependent lymphocytotoxicity (n=49; PRA=84.1+/-12.1%) and antihuman globulin (n=60; PRA=92.5+/-5.8%) groups, the 3-month graft survivals were 28% and 30% lower for unacceptable residue mismatches. CONCLUSIONS: These studies underscore the importance of a comprehensive serum screening analysis in the selection of appropriately mismatched donors for highly sensitized transplant patients. PMID- 9210499 TI - Monocytes-macrophages and cytokines/chemokines in fine-needle aspiration biopsy cultures: enhanced interleukin-1 receptor antagonist synthesis in rejection-free kidney transplant patients. AB - BACKGROUND: Monocytes-macrophages are found within kidney allografts during the first days after surgery, where they perform "housekeeping" tasks, participate in postreperfusion injury, and act as antigen-presenting cells, as well as become involved in the effector phase of acute rejection. They also seem to play a prominent role in chronic rejection. We quantified their presence in fine-needle aspiration biopsies and studied the growth factors that, we hypothesized, would mark the different implications of the presence of monocytes-macrophages. METHODS: Fine-needle aspiration biopsies were obtained from 56 adult renal transplants and analyzed for CD14+ using the alkaline phosphatase-anti-alkaline phosphatase procedure. Thirty-three patients were studied on the production of interleukin (IL)-1 receptor antagonist (IL-1ra), IL-6, IL-8, macrophage colony stimulating factor, monocyte chemotactic protein 1, and macrophage inflammatory protein 1alpha by aspiration biopsies cultures using enzyme-linked immunosorbent assay techniques. RESULTS: CD14+ cells were present at significantly higher numbers in steroid-resistant acute rejections but also during the first days after surgery, especially if acute tubular necrosis was present. We found a significantly higher production of IL-1ra by rejection-free patients compared with acutely rejecting patients, and this difference was already established on day 7 after surgery (10+/-10.5 days before rejection). CONCLUSIONS: Monocytes macrophages are present at higher numbers in aspiration biopsies of kidney transplant patients suffering either acute tubular necrosis or steroid-resistant rejections, but they are present during the first days after transplant in stable patients, too. The production of IL-1ra is significantly up-regulated in stable patients, which suggests that monocytes-macrophages may constitute an early key factor in the down-regulation of the anti-allograft immune response. PMID- 9210500 TI - Cell-mediated cytotoxicity: a predictor of chronic rejection in pediatric HLA haploidentical renal transplants. AB - BACKGROUND: Recipient antidonor cytotoxic T-cell activity has been associated with graft loss and acute rejection in renal allograft recipients. The role of immunologic mechanisms in the development of chronic graft rejection is controversial. We analyzed all living related renal transplants performed at Children's Hospital (Boston, MA) from 1983 to 1995 to assess whether cell mediated cytotoxicity, determined in vitro and measured before transplantation, was predictive of chronic rejection. METHODS: Eighty-three patients were studied retrospectively. Fifty-seven patients with one haplotype-matched renal transplants from living related donors were studied to determine the association between cell-mediated lympholysis (CML) level, acute rejection, chronic rejection, and graft failure. Acute rejection was defined by the decision to treat. Chronic rejection was defined by histology and/or the absolute serum creatinine value using an increasing serum creatinine level >1.0 mg/dl for children less than 3, a creatinine level >1.5 mg/dl for children between 3 and 10 years of age, and a creatinine level >2.0 mg/dl for children above 10 years of age. Return to dialysis or retransplantation was considered graft failure. RESULTS: Of the 57 haploidentical patients, there were 33 males and 24 females. The mean age at transplant was 11.1 years (SD=6.7). Twelve patients developed chronic rejection, 24 patients developed acute rejection, and 7 patients had graft failure. Pretransplant cytotoxic T lymphocyte activity was associated with chronic rejection (P=0.001) and graft failure (P=0.013) but only marginally with acute rejection (P=0.058). Controlling for age and sex, Cox's proportional hazards model revealed that CML level was predictive of time to chronic rejection (P<0.01) but not acute rejection (P=0.11). It was estimated that every 1-unit increase in CML level raises the monthly risk of chronic rejection by 7%. Ten children received HLA-identical kidneys from their siblings. There were no episodes of chronic rejection after 5 years. Two patients with high CML levels had episodes of acute rejection; both patients responded to treatment. CONCLUSION: Our data demonstrate an association between pretransplant cell mediated cytotoxicity and the occurrence of chronic rejection in living related one-haploidentical renal transplants in pediatric patients. PMID- 9210501 TI - Pharmacokinetics of an oral solution of the microemulsion formulation of cyclosporine in maintenance pediatric liver transplant recipients. AB - BACKGROUND: A comparison of the oral bioavailability of cyclosporine from the original formulation (CsA) and from the new formulation, cyclosporine for microemulsion (CsA-ME), was made in pediatric maintenance liver transplant patients within two age groups (group 1, ages 1-5 years; group 2, ages 6-17 years) in an open-label, multicenter, randomized crossover trial. All patients were at least 6 months past transplantation and were receiving CsA maintenance therapy. METHODS: In study period 1 (days 1 through 14), patients were administered either CsA or CsA-ME at the same b.i.d. dosage as their maintenance therapy. Upon entry into period 2 (days 15 through 28), patients were converted to the alternate formulation at a 1:1 mg dose ratio. On day 29, all patients returned to the CsA treatment administered at study entry, with follow-up on day 35. Dosage adjustments were not allowed with either CsA or CsA-ME. Twelve-hour pharmacokinetic profiling was performed at the end of periods 1 and 2. RESULTS: Both the mean area under the concentration-versus-time curve and the mean maximum blood concentration of cyclosporine-both normalized for dose-were significantly increased: by 66% and 109%, respectively, in patients receiving CsA-ME compared with those receiving CsA in group 1 and by 39% and 75%, respectively, in group 2. During this study, liver function remained stable, and serum creatinine and blood pressure did not differ significantly between treatment groups. CONCLUSIONS: This study shows increased bioavailability in all patients converted to CsA-ME, with the greatest increase seen in patients with the lowest initial cyclosporine bioavailability. The tolerability was similar between the two formulations during this study. PMID- 9210502 TI - Kidney function after 1:1 conversion to the cyclosporine microemulsion formulation in children with liver allografts. AB - BACKGROUND: One-to-one (mg:mg) conversion from the conventional to the microemulsion formulation of cyclosporine (CsA) is advocated as a simple way to use the new therapeutic regimen. However, the potentially harmful effects of the conversion on kidney function in nonrenal transplant recipients are poorly known. METHODS: Renal effects of the conversion were prospectively investigated in 22 pediatric liver transplant recipients (mean age, 8.4 years; mean time from transplantation, 3.2 years). Patients were followed for 12 months. Pharmacokinetic studies were performed at baseline and 5 days and 6 and 12 months after conversion. RESULTS: Peak concentration, minimum concentration, average steady state concentration, and area under the concentration-versus-time curve increased by 60-130% after conversion. Graft losses, progressive deterioration of graft function, and acute rejection episodes did not occur. The mean glomerular filtration rate (GFR) was 103 ml/min/1.73 m2 at baseline and 100 ml/min/1.73 m2 after 12 months. However, 6 of the 22 patients showed at least a 15% (range, 16 38%) decrease in GFR between baseline and 6 months (P<0.01). They had a significantly higher increase in average steady state concentration between baseline and 6 months than the six patients with the best outcome in GFR during the same time period (164 ng/ml vs. 53 ng/ml, P<0.05). At this point (6 months), target CsA trough levels were reduced by 20-30%, while the mean area under the concentration-versus-time curve remained above that obtained at baseline. The GFR of three of the six patients subsequently improved. CONCLUSIONS: One-to-one conversion can be performed safely in liver transplant recipients if strict follow-up is feasible. PMID- 9210503 TI - A prospective randomized trial comparing interleukin-2 receptor antibody versus antithymocyte globulin as part of a quadruple immunosuppressive induction therapy following orthotopic liver transplantation. AB - BACKGROUND: Quadruple immunosuppressive induction therapy has been shown to markedly reduce the incidence of acute rejection episodes without increasing the incidence of infectious complications after liver transplantation. However, the use of polyclonal antibody preparations (e.g. antithymocyte globulin [ATG]) is associated with side effects such as fever and tachycardia. To evaluate the efficacy and the safety of a monoclonal antibody directed against the interleukin 2 receptor (BT563) in comparison with ATG as part of a quadruple induction regimen, a prospective, randomized study was conducted. METHODS: Eighty consecutive adult recipients of primary orthotopic liver transplants were randomized to receive either BT563 (10 mg/day; days 0-12; n=39) or ATG (5 mg/kg/day; days 0-6; n=41) in addition to the standard immunosuppressive protocol consisting of cyclosporine, and prednisolone, and azathioprine. RESULTS: Patients treated with BT563 had a significantly lower incidence of steroid-sensitive rejection episodes (3 vs. 11; P<0.025) and also significantly fewer drug-related side effects (4 vs. 18, P<0.038) when compared with patients treated with ATG. The incidence of infectious complications was not different between the two groups. Patient survival did not differ significantly between the two groups (84.6% at 1, 2, and 3 years in the BT563 group and 90.2% at 1 year and 87.8% at 2 and 3 years for the ATG group). Analysis of graft function showed an advantage for the BT563 group in terms of postoperative bilirubin levels. However, no differences were observed in long-term follow-up between the two groups. CONCLUSIONS: Our results indicate that treatment with anti-interleukin-2 receptor antibody as part of quadruple induction therapy after orthotopic liver transplantation is safe and effective and shows fewer steroid-sensitive rejection episodes as well as fewer side effects when compared with quadruple induction therapy including ATG. PMID- 9210504 TI - Interstitial pneumonitis in bone marrow transplant recipients is associated with local production of TH2-type cytokines and lack of T cell-mediated cytotoxicity. AB - BACKGROUND: Interstitial pneumonitis, especially associated with cytomegalovirus (CMV) infection, is a serious complication after bone marrow transplantation (BMT), with a high fatality rate despite adequate antiviral treatment. The aim of this study was to elucidate the local immunopathogenesis of interstitial pneumonitis caused by CMV or other agents in BMT recipients. METHODS: Cryopreserved lung tissue obtained from 12 patients with interstitial pneumonitis following BMT was analyzed for cytokine production at the single-cell level using a cytokine-specific monoclonal antibody and immunohistochemical technique. Cytokine production in individual cells was analyzed using monoclonal antibodies to 23 different human cytokines: interleukin (IL)-1 to IL-13, tumor necrosis factor (TNF)-alpha, TNF-beta, interferon-gamma (IFNgamma), granulocyte colony stimulating factor, granulocyte-macrophage colony-stimulating factor, and transforming growth factor (TGF)-beta1 to 3. RESULTS: Marrow transplant patients with interstitial pneumonia had increased numbers of infiltrating alveolar macrophages, CD3+, CD4+ T cells, and CD40+ B cells and significantly increased numbers of IL-4-, IL-10-, IL-1-, TGF-beta1-, TGF-beta2-, and TGF-beta3-producing cells than controls. IL-2-, IFN-gamma-, and TNF-beta-producing cells were undetectable in most patients with CMV pneumonitis (n=7). Neither perforin positive CD8+ T lymphocytes nor up-regulation of the apoptotic pathway was detected in lung tissue from patients with interstitial pneumonia. In contrast, extensive local production of IgA, IgG, and IgM was demonstrated in all patients. Intracellular and extensive extracellular deposition of CD68, the L-1 antigen synthesized in CD14+ macrophages, was found. CONCLUSIONS: The cytokine profile suggested that Th1-type cytokine production was absent, whereas production of Th2 type cytokines was significantly up-regulated. Interstitial pneumonitis in BMT recipients with fatal outcome (11/12 patients) was associated with dysregulation in the local cytokine network notable for a predominant Th2 immune response with minimal or absent T cell-mediated cytotoxicity. PMID- 9210505 TI - Results of a survey of infectious disease testing practices by organ procurement organizations in the United States. AB - BACKGROUND: Information related to infectious disease testing policies and practices of organ procurement organizations in the United States does not currently exist. METHODS: A total of 63 organ procurement organizations in the United States were surveyed during May 1996. Participants responded to a detailed questionnaire concerning infectious disease tests performed for tissue and solid organ donors and policies related to the reporting and notification of positive test results. RESULTS: The response rate was 77.8%. The majority of testing is performed by hospital laboratories with an expected turnaround time of 5 hr or less by 71% of organ procurement organizations. Almost all routinely perform screening tests for human immunodeficiency virus, hepatitis C virus, cytomegalovirus, syphilis, human T lymphocyte virus I, and hepatitis B surface antigen. Other tests are performed with greater variability. Although the majority of organ procurement organizations perform confirmatory tests when screening tests are positive, 35% do not perform confirmatory testing or do so only sporadically. There are a wide range of policies concerning the subsequent reporting of positive infectious disease tests and to whom results should be reported. CONCLUSIONS: Infectious disease testing policies of organ procurement organizations, particularly for solid organs, demonstrate variability in interpretation and perceived significance of positive test results, the initiation or need for reflex and confirmatory testing, the reporting of positive results, and to whom positive test results should be reported. There is a need for a consistent national policy for appropriate infectious disease testing and reporting of results. PMID- 9210506 TI - A dual-marker system for quantitative studies of myoblast transplantation in the mouse. AB - BACKGROUND: Myoblast transplantation (MT) is a potential approach for gene transfer into skeletal muscle, the efficiency of which depends upon the number of copies of donor genome incorporated into the host tissue. We have developed a system for quantitative studies of MT that measures amounts of donor-derived genome in host muscles and estimates the contributions of donor cell survival and proliferation in vivo. METHODS: [14C]thymidine-labeled, male myoblasts were transplanted into female muscles, providing two donor cell markers, Y chromosome and [14C]. The markers were measured in muscle extracts by slot blotting and scintillation counting, respectively. RESULTS: In each extract, the amount of Y chromosome was used to quantify donor-derived genome, whereas the radiolabel provided an estimate of cell survival. Furthermore, the different modes of inheritance of the markers meant that proliferation of surviving donor cells was detected as a change in marker ratio. CONCLUSIONS: This system provides a method for assessing potential improvements of MT. PMID- 9210508 TI - Comparison of a new quantitative cytomegalovirus DNA assay with other detection methods. AB - OBJECTIVE: We assessed a new cytomegalovirus (CMV) DNA hybridization assay. We also compared the assay with other currently used assays to determine its use in the early detection of active CMV infection. PATIENTS AND METHODS: Sequential whole blood samples collected from 109 patients who had undergone orthotopic liver transplantation were tested using the Murex hybrid capture system, cell culture, antigen detection, and serology. Liver biopsies performed during the study period for graft dysfunction in 84 patients were examined for histological features of CMV hepatitis. The biopsies were also immunostained for the presence of CMV antigens. RESULTS: Fifteen patients developed clinically significant CMV disease (CMV syndrome in six patients and CMV hepatitis in nine patients, including two patients with disseminated CMV disease). In all 15, CMV DNA was detected by the hybrid capture assay between 1 and 20 days before other CMV assays. Fourteen of the 15 patients had CMV DNA levels greater than 50 pg/ml; the other patient had a value of 48 pg/ml. Of the remaining 94 patients with no evidence of CMV disease, 86 were negative by the hybrid capture assay and 8 were positive; all but one patient had values less than 50 pg/ml. DNA levels fell rapidly in all patients during antiviral therapy. CONCLUSION: Unlike conventional CMV detection methods, this hybridization assay is an early predictor of clinically significant CMV infection after liver transplantation and also provides quantitation of viral load, allowing monitoring of antiviral therapy. PMID- 9210507 TI - Standardization of procedure for efficient ex vivo gene transfer into porcine pancreatic islets with cationic liposomes. AB - BACKGROUND: New strategies to improve the outcome of encapsulated porcine islet transplantation may involve the transfer of gene sequences affecting islet viability. While adenoviral vectors appear as the most efficient gene transfer system so far established for islets, non-viral-based vectors are most likely to fulfill microbiological safety criteria and be retained in the clinical setting. Our aim was to standardize the procedures of gene transfer into adult porcine islets using cationic liposome DOTAP. METHODS: Porcine islets obtained by collagenase digestion and density gradient purification were lipofected with plasmids coding for luciferase or beta-galactosidase under the control of simian virus 40 or cytomegalovirus promoter. The following parameters were explored: exposure time to vector (1-48 hr), DNA amount (1-15 microg/500 islets), and DOTAP to DNA ratio (2-16). Reporter gene expression was determined 48-72 hr after lipofection. RESULTS: Efficiency and reproducibility of transfection were maximal with the following procedure: 3-hr exposure time followed by islet washing, 12 microg of DNA per 500 islets (150 microm equivalent), and DOTAP to DNA ratio of 12 microl/microg. Freshly isolated islets in large aliquots (n=4000 in 50-ml tubes) were efficiently transduced with this procedure, and distribution of gene expression was homogenous when islets were subsequently plated in 500-islet aliquots. Luciferase gene expression was detected for a minimum of 7 days after lipofection. Gene expression was also evident up to 4 weeks after islet transplantation beneath the kidney capsule of athymic mice. Transfection of islets using the beta-galactosidase vector yielded 25% positive islets. Islet viability was not adversely affected. CONCLUSIONS: This islet lipofection procedure may help achieve the local release of a bioactive peptide in the graft environment and have therapeutic applications in islet transplantation. PMID- 9210509 TI - Expression of chemokine genes during rejection and long-term acceptance of cardiac allografts. AB - Chemokines are cytokines with chemoattractant properties for leukocytes. They may play a critical role in directing leukocytes to graft sites and in amplifying intragraft inflammation during rejection. Previous studies have tested the intragraft expression of chemokine genes during the rejection of allogeneic skin grafts in mice. In the current study, we used a heterotopic heart transplant model in mice to test the intragraft expression of these genes in nonrejecting cardiac isografts, rejecting cardiac allografts, and cardiac allografts that were accepted due to immunosuppression with gallium nitrate. With the exception of low levels of interleukin-1beta and JE, intragraft expression of the the proinflammatory cytokine genes was not observed in either isografts or native heart. Two distinct patterns of chemokine mRNA were observed in the rejecting cardiac allografts. Intra-allograft expression of interleukin-1beta, interferon gamma-inducible protein, JE, and KC was prominent by day 3 after transplantation. The expression of macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and regulated upon activation, normal T cell expressed and secreted (RANTES) was at low or undetectable levels at day 3 after transplantation but at high levels by day 8 after transplantation. Sixty days after transplantation, intra-allograft expression of chemokines in hearts from gallium nitrate-treated recipients indicated low levels of MIP-1alpha, MIP-1beta, and KC but high levels of interferon-gamma-inducible protein and RANTES. PMID- 9210510 TI - Use of the methylxanthine derivative A802715 in transplantation immunology: I. Strong in vitro inhibitory effects on CD28-costimulated T cell activities. AB - BACKGROUND: Recently, methylxanthines such as pentoxifylline (PTX) were shown to be immunosuppressive in vitro. Unfortunately, when used in transplant patients, PTX was poorly active as an immunosuppressant. Here we report that the new methylxanthine derivative A802715 not only is more active than PTX, it also suppresses the cyclosporine (CsA)-resistant "signal two"-dependent pathway of T cell proliferation, making it an interesting drug to associate with CsA. METHODS: "Signal one"- and "signal two"-dependent T cell activation was investigated with purified human T cells stimulated with immobilized anti-CD3 or anti-CD28 monoclonal antibody (mAb) plus phorbol myristate acetate (PMA) or with a 3T6 mouse fibroblast cell line presenting anti-CD3 mAb on transfected human Fcgamma receptors II (FcgammaRII) in the presence or absence of transfected B7-1 (CD80) molecules. RESULTS: A802715 was more immunosuppressive in the mixed lymphocyte reaction (MLR) than PTX. A802715 dose-dependently suppressed polyclonal signal one-dependent T cell activation induced by anti-CD3 mAb/PMA. In addition, A802715 also suppressed signal two-dependent T cell proliferation induced by anti-CD28 mAb/PMA. The expression of the interleukin-2 receptor on T cells stimulated by anti-CD3 mAb presented on 3T6/FcgammaRII cells was equally well suppressed by A802715 and PTX. In contrast, interleukin-2 receptor or CD40L (gp39) expression by T cells after stimulation with the same anti-CD3 mAb- 3T6/FcgammaRII cells, but coexpressing transfected B7-1, was only suppressed by A802715. The anticipated synergism between A802715 and CsA was confirmed in MLR assays. Moreover, generation of cytotoxic T lymphocytes during MLR with Epstein-Barr virus-transformed B cells, which strongly express B7-1 and B7-2, was also inhibited by A802715. CONCLUSIONS: These in vitro data indicate that the A802715 (1) is a stronger immunosuppressant for T cells than PTX, (2) suppresses T cell activation pathways that are resistant to PTX or CsA, and (3) acts synergistically with CsA. PMID- 9210511 TI - Abnormal differentiation of thymocytes induced by free cyclosporine is avoided when cyclosporine bound to N-(2-hydroxypropyl)methacrylamide copolymer carrier is used. AB - BACKGROUND: The side effects of cyclosporine (CsA)-including nephrotoxicity and abnormal differentiation of thymocytes developing in the thymus-can be decreased or even avoided using targeted conjugates of CsA, where both targeting moiety and drug are bound to water-soluble polymeric carrier based on N-(2-hydroxypropyl) methacrylamide (HPMA). METHODS: Irradiated, syngeneic bone marrow transplanted mice (BALB/c and A/Ph) were treated intraperitoneally for 4 weeks with 20 mg/kg of free CsA, HPMA-conjugated CsA, or antibody-targeted HPMA-bound CsA. Immunohistology of the thymus was performed together with two-color flow cytometry to detect the effect of different forms of CsA on individual thymocyte subpopulations. RESULTS: . We have shown that free CsA strongly abrogated T-cell development. The appearance of mature thymocytes expressing CD3(high) is almost completely inhibited (1.8%) after free CsA treatment, whereas these cells are well detectable in controls (22%) and HPMA polymer-bound CsA-treated animals (19%). Immunohistological studies have shown acellular rests of the medulla after free CsA treatment, whereas well-stained medullary thymocytes were detected in controls and after exposure to antibody-targeted HPMA. conjugated CsA. CONCLUSIONS: HPMA-conjugates of CsA are generally more specific in their targeting to T lymphocytes. It was found that nonspecific binding of CsA to erythrocytes and plasma lipoproteins is significantly reduced using anti-CD3 targeted, HPMA polymer-bound CsA In addition, the entry of these macromolecules into the thymus is limited-probably due to the blood-thymus barrier-and HPMA conjugates of CsA, unlike free drug, do not abrogate T-cell development in bone marrow transplanted mice. PMID- 9210512 TI - Detection of HLA class I- and class II-specific antibodies by flow cytometry and PRA-STAT screening in renal transplant recipients. AB - BACKGROUND: Screening for HLA-specific antibodies has been performed by complement-dependent lymphocytotoxicity for many years. In recent years, methods involving the use of flow cytometry or ELISA have been developed. METHODS: This study has compared a flow cytometric screening technique for the detection of HLA class I- and class II-specific antibodies with a commercially available ELISA technique, PRA-STAT. RESULTS: A significant correlation was found between the two methods for the detection of antibodies in patients after transplantation (P<0.001). Specificity analysis confirmed that the PRA-STAT technique detected both HLA class I- and class II-specific antibodies. Screening of serum samples from patients who experienced graft loss by cytotoxic, flow cytometric, and PRA STAT techniques showed that there was a significant correlation between all three methods for the detection of antibody, but that the best correlation for the panel-reactive antibody level was that between the flow cytometric and PRA-STAT techniques (r=0.86). This was principally due to the detection of both HLA class I- and class II-specific antibodies by these methods, whereas cytotoxic screening detected only class I-specific antibodies. CONCLUSIONS: These results suggest that PRA-STAT is a useful technique for the detection of both HLA class I- and class II-specific antibodies, rather than only class I-specific antibodies as previously described. PMID- 9210513 TI - Recognition of major histoincompatibilities after transplantation with marrow from HLA closely matched donors. AB - BACKGROUND: To determine the extent to which major histoincompatibilities are recognized after bone marrow transplantation, we characterized the specificity of the cytotoxic T lymphocytes isolated during graft-versus-host disease. We studied three patients transplanted with marrow from donors who were histoincompatible for different types of HLA antigens. METHODS: Patient 1 was mismatched for one "ABDR-antigen" (HLA-A2 versus A3). Two patients were mismatched for antigens that would usually not be taken into account by standard selection procedures: patient 2 was mismatched for an "HLA-A subtype" (A*0213 versus A*0201), whereas patient 3 was mismatched for HLA-C (HLA-C*0501 versus HLA-C*0701). All three HLA class I mismatches were detected by a pretransplant cytotoxic precursor test. RESULTS: Analysis of the specificity of the cytotoxic T lymphocyte clones isolated after transplantation showed that the incompatibilities detected by the pretransplant cytotoxic precursor assay were the targets recognized during graft-versus-host disease. CONCLUSIONS: Independent of whether the incompatibility consisted of a "full" mismatch, a "subtype" mismatch, or an HLA-C mismatch, all clones recognized the incompatible HLA molecule. In addition, some of these clones had undergone antigen selection and were clearly of higher specificity than the ones established before transplantation, indicating that they had been participating directly in the antihost immune response. PMID- 9210514 TI - Atrioventricular block after administration of atropine in patients following cardiac transplantation. AB - BACKGROUND: Atropine is widely used as a parasympatholytic agent during diagnostic and therapeutic procedures. We observed an unexpected paradoxical response to atropine after cardiac transplantation. METHODS: In a study investigating the occurrence of autonomic reinnervation after cardiac transplantation, atropine, at 0.015 mg/kg body weight, was given intravenously to 23 patients (mean age, 56+/-8 years) 98 days to 6.4 years after transplantation. RESULTS: Two patients experienced a witnessed syncope 40 and 150 min after administration of atropine. Second-degree atrioventricular (AV) block was documented in the first patient immediately afterward, and third-degree AV block was seen on 24-hr electrocardiogram monitoring in the second patient. A third patient developed documented AV block 15 min after atropine but experienced no sequelae because of a previously implanted pacemaker. CONCLUSIONS: Although the underlying mechanism is not clear, these findings suggest that atropine may paradoxically cause high-degree AV block in patients after transplantation. Accordingly, it should be used with caution and appropriate monitoring in these patients. PMID- 9210515 TI - Effect in supralethally irradiated rats of granulocyte colony-stimulating factor and lisofylline on hematopoietic reconstitution by syngeneic bone marrow or whole organ passenger leukocytes. AB - We have previously shown the existence of migratory hematopoietic stem cells in adult solid organs. This study demonstrates that granulocyte colony-stimulating factor (G-CSF) and lisofylline, a phosphatidic acid inhibitor that suppresses hematopoiesis-inhibiting cytokines, can enhance the engraftment of organ-based hematopoietic stem cells. When syngeneic heart grafts or liver nonparenchymal cells were transplanted into lethally irradiated (9.5 Gy) Lewis rats, complete hematopoietic reconstitution and animal survival were significantly improved by treating the recipient with G-CSF or, to a lesser extent, with lisofylline. Pretreatment of hepatic nonparenchymal cell donors with G-CSF, but not lisofylline, also resulted in striking improvement of recipient survival which was associated with an augmented subpopulation of donor stem cells. The results suggest that these drugs can be used to enhance the chimerism that we postulate to be the basis of organ allograft acceptance. PMID- 9210516 TI - Biophysical aspects of liver aeration by vascular persufflation with gaseous oxygen. AB - BACKGROUND: Venous systemic oxygen persufflation of the liver (i.e., gaseous insufflation of oxygen via the venous vascular system) has proven to be an effective tool for preventing anoxic tissue injury during extended time periods of ischemic preservation. It also allows for an improved recovery of the persufflated organ after orthotopic transplantation. METHODS: Biophysical aspects of the persufflation technique with regard to persufflation pressure (9 mmHg versus 18 mmHg) and oxygen concentration (pure oxygen versus air) in the persufflation gas were investigated in rat livers, using epi-illumination microscopic detection of autofluorescence of NADH, which accumulates in anoxic tissue. RESULTS: We demonstrated that a low-pressure persufflation (9 mmHg) is as sufficient as a higher pressure persufflation (18 mmHg) in oxygenating the ischemic organ. Moreover, oxygenation of the liver was found to be complete and rather homogeneous upon the pure oxygen persufflation, irrespective of the insufflation pressure used. In contrast, insufflation of air instead of pure oxygen resulted in insufficient aeration of the liver, even at the higher persufflation pressure of 18 mmHg. CONCLUSIONS: Our results indicate that the oxygen concentration of the persufflation gas rather than the persufflation pressure is a determinant of successful tissue oxygenation during cold storage. PMID- 9210517 TI - Low HLA-DR expression on monocytes as a prognostic marker for bacterial sepsis after liver transplantation. AB - BACKGROUND: Low HLA-DR expression on monocytes is associated with an increased risk of infection after surgery or trauma. We determined the value of this parameter as a marker for sepsis after liver transplantation. METHODS: The percentage of monocytes expressing HLA-DR was determined by flow cytometry before and after liver transplantation in nine patients. Five lung and 20 kidney transplant recipients served as controls. RESULTS: Bacterial sepsis occurred in 5 of 9 liver transplant patients and 0 of 24 control patients. Monocyte HLA-DR expression decreased <50% in all five patients with sepsis. HLA-DR expression dropped before (n=4) or at the time of sepsis (n=1), and remained low for 13 weeks. HLA-DR expression remained >50% in the four liver transplant patients without sepsis. Only 1 of 25 control patients had persistently low monocyte HLA DR expression. CONCLUSIONS: Monitoring of monocyte HLA-DR expression may be helpful in identifying liver transplant patients who have an increased risk of imminent bacterial sepsis. PMID- 9210518 TI - AP-1 mediates trans-synaptic induction of tyrosine hydroxylase gene expression in adrenal medulla but not in superior cervical ganglia. AB - Reserpine treatment leads to a rapid trans-synaptic increase of the tyrosine hydroxylase (TH) gene transcription rate and mRNA levels in catecholaminergic tissues including the adrenal medulla (AM) and the superior cervical ganglia (SCG). In the AM, the formation of a specific protein complex with the TPA responsive element located in the proximal region of the TH gene was enhanced between 30 min and 8 hr following the injection. This complex appears to contain a member of the Fos family and an antigenically related Jun protein. Moreover, the prolonged and enhanced expression of the c-Fos protein in the AM and its phosphorylation are likely to contribute to the increased TH transcription following reserpine treatment. Most strikingly, in the SCG, the trans-synaptic induction of TH transcription is transduced by totally different mechanisms, since no AP-1 complex and only minute amounts of c-Fos immunoreactivity were detected. Our study provides the first demonstration that, following the same stimulus, the induced expression of a single gene is mediated by different cis- and trans-acting factors in two distinct tissues sharing the same embryonic origin. PMID- 9210519 TI - Effects of excitatory amino acid antagonists on dendrotoxin-induced increases in neurotransmitter release and epileptiform bursting in rat hippocampus in vitro. AB - Alpha-dendrotoxin (alpha-DTx), a snake venom toxin which blocks several types of fast-activating voltage-dependent potassium channels, induces limbic seizures and neuronal damage when injected into the brain. The mechanisms underlying these convulsant and neuropathological actions are not fully understood. We have studied the effects of alpha-DTx on neurotransmitter release and electrical activity in rat hippocampal brain slices and the role of excitatory amino acid receptors in mediating these actions of the toxin. alpha-DTx increased the basal release of acetylcholine, glutamate, aspartate, and GABA in a concentration dependent manner and induced epileptiform bursting in the CA1 and CA3 regions of the slice. The increase in neurotransmitter release was evident during the first 4 min after toxin addition, whereas the bursting appeared after a concentration dependent delay (20-40 min with 250 nM toxin). The N-methyl-D-aspartate (NMDA) receptor antagonists AP5 and MK-801 had no effect on the frequency or amplitude of dendrotoxin-induced epileptiform bursts, but the non-NMDA antagonists CNQX and DNQX abolished bursting in both CA1 and CA3 within 4-6 min. In contrast, the toxin-induced increases in neurotransmitter release were not blocked by DNQX. This study has demonstrated that, following exposure to alpha-DTx, there is a rapid increase in the release of neurotransmitters which precedes the onset of epileptiform bursting in the hippocampus. Since DNQX abolished the bursting but had no effect on the increase in neurotransmitter release, these results suggest that DNQX blocks alpha-DTx-induced epileptiform activity by antagonism of postsynaptic non-NMDA receptors. PMID- 9210520 TI - Purification and cDNA cloning of mouse BM89 antigen shows that it is identical with the synaptic vesicle protein synaptophysin. AB - The BM89 antigen, first identified in porcine brain by means of a monoclonal antibody, is a neuron-specific molecule widely distributed in the mammalian central and peripheral nervous system (Merkouri and Matsas: Neuroscience 50:53 68, 1992). Here we describe the purification of BM89 antigen from porcine and mouse brain by immunoaffinity chromatography using, respectively, the previously described BM89 monoclonal antibody which belongs to the IgM class and a specific polyclonal antibody generated in the present study. This antibody was also used for the cDNA cloning of the BM89 antigen from mouse brain. cDNA sequencing revealed that the mouse BM89 antigen is identical with the synaptic vesicle protein synaptophysin which is implicated in the control of regulated exocytosis and neurotransmitter release. Mouse BM89 antigen/synaptophysin exhibits, except for one extra amino acid, 100% identity with rat synaptophysin and substantial sequence identity with bovine (92.5% identity) and human (94.8% identity) synaptophysin, but only 59.8% identity with Torpedo synaptophysin. Northern and Western blot analyses confirmed that the mouse BM89 antigen/synaptophysin is expressed only in neural tissues. PMID- 9210521 TI - Triton-soluble phosphovariants of the heavy neurofilament subunit in developing and mature mouse central nervous system. AB - The low abundance of soluble neurofilament (NF) subunits in mature axons has suggested that newly synthesized NF proteins rapidly assemble into highly stable polymers and associate with the Triton X-100-insoluble cytoskeleton. The dynamic nature of these subunit associations in vivo remains unresolved, and the applicability of this assembly model to NFs in other neuronal compartments or to developing neurons is unknown. Here, we report that a unique pool of Triton X-100 soluble, extensively phosphorylated, high molecular weight NF subunits (NF-H, or H-200) are abundantly expressed in the mouse CNS during early postnatal development and persist in the perikaryal compartment of some mature neurons. Triton-soluble H-200 subunits appeared at postnatal day 14 (P14) and remained high through P60, beyond which the percentage declined to marginal levels by P120. Medium and low molecular weight NF (NF-M and NF-L, respectively) were at all times only detectable within the cytoskeleton. Comparison of soluble and cytoskeleton-associated H-200 immunoreactivity indicated that certain phosphorylation-dependent epitopes were confined to the cytoskeleton. Pulse-chase radiolabeling analyses in optic pathway demonstrated that some Triton-soluble NF H subunits are extensively phosphorylated within retinal perikarya before they are incorporated into Triton-insoluble structures. These findings indicate that the assembly behaviors of NF-H differ substantially from those of NF-M and NF-L, and that the interaction of NF-H with NFs may be more dynamic than is generally recognized, especially during brain development and within specific compartments of mature neurons. PMID- 9210522 TI - Functional recovery from sciatic nerve crush lesion in the rat correlates with individual differences in responses to chronic intermittent stress. AB - The aim of the present study was to monitor the influence of chronic stress on functional recovery from a sciatic nerve crush lesion in the rat. Male Wistar rats underwent standard unilateral sciatic nerve crush. Subsequently, chronic stress was induced during the recovery phase using a daily 30 min shock box session where rats received three electric footshocks each session (0.5 sec, 1 mA). Reduced body weight gain, adrenal gland hypertrophy, and thymus involution indicated that the stress rats were chronically stressed. Evaluation of sensorimotor function revealed significant differences in recovery between control and stress groups. Correlational analysis of individual stress rats indicated that recovery of the walking pattern was negatively correlated with adrenal gland and medulla enlargement, thymus involution, and plasma levels of adrenocorticotrophic hormone (ACTH) and corticosterone 45 min following the final stress session. In control rats, the index of sciatic nerve function (SF index, expressed as the difference between the injured paw and the intact contralateral paw as a percentage) was significantly correlated with adrenal medulla weight only. The present study reveals that chronic intermittent footshock stress impedes sensorimotor recovery following a sciatic nerve crush lesion and that the consequences of chronic intermittent stress are individually determined. We suggest that the quality of functional locomotor recovery after nerve crush lesion is related to the adaptive capacity or coping style of the individual rat. PMID- 9210523 TI - Association of c-fos mRNA expression and excitotoxicity in primary cultures of mouse neocortical and cerebellar neurons. AB - The effect of excitatory amino acids (EAAs) on c-fos mRNA expression was studied in primary cultures of mouse cerebellar granule cells and in neocortical neurons after 2 and 7 days in vitro (div). In cultured granule cells at 2 and 7 div, and in cortical neurons at 2 div, exposure to low levels (< or = 10 microM) of a variety of EAAs (viz. glutamate [Glu], S-sulpho-L-cysteine [SC], N-methyl-D aspartate [NMDA], alpha-amino-3-hydroxy-5-methyl-4-isoxazole [AMPA], and kainate [KA]) resulted in a transient increase in the level of c-fos mRNA which peaked at 30 min but returned to a basal level by 120 min. However, exposure of granule cells (7 div) to high levels (250 microM) of Glu, NMDA, KA, SC and of cortical neurons (7 div) to high levels (250 microM) of Glu, NMDA, KA, SC, or AMPA and to low levels (< or = 10 microM) of Glu and AMPA resulted in a delay in c-fos mRNA induction but a subsequent, progressive increase that was sustained for at least 240 min. Furthermore, this effect was accompanied by a dose-related increase in the release of the cytosolic enzyme, lactate dehydrogenase, used as an indicator of excitotoxicity. A ratio (Q240/30) for the steady-state levels of c-fos mRNA after 30 min and 240 min of exposure to EAAs was determined which showed that Q240/30 >2 correlated reproducibly with excitotoxic cell death, whereas a ratio of < or = 1 correlated with a nonexcitotoxic event. In both cell types at 7 div, coadministration of the selective NMDA receptor antagonist, DL(+/-)-2-amino-5 phosphonopentanoic acid (APV) with cytotoxic levels of Glu 1) protected against EAA-induced neurotoxicity and 2) exhibited a transient c-fos mRNA expression (Q240/30 values approximately 1). In contrast, the AMPA/KA receptor antagonist, 6 cyano-7-nitroquinoxaline-2,3-dione (CNQX), provided no protection against excitotoxicity and had no significant effect on the Glu-induced delay in c-fos mRNA expression. These results suggest that the Q240/30 c-fos mRNA ratio may 1) be used as a predictive index for excitotoxic neuronal death, 2) provide information on the identity of the receptor subtype mediating excitotoxicity in different brain cell types, and 3) aid in establishing the role of excitotoxicity during the development of neurons in vitro. PMID- 9210524 TI - Restriction of microM-calcium-requiring calpain activation to the plasma membrane in human neuroblastoma cells: evidence for regionalized influence of a calpain activator protein. AB - Regulation of the microM-calcium-requiring form of calpain (mu calpain) was studied in SH-SY-5Y human neuroblastoma cells. Immunoblot analysis demonstrated that the vast majority of mu calpain is localized within cytosolic pools. Calpain activation was monitored as a function of autolysis within intact cells following calcium influx from the culture medium by calcium ionophores A23187 or ionomycin, or following release of calcium from intracellular stores by thapsigargin. Within intact neuronal cells, following an influx of calcium into the cytosolic from either extracellular or intracellular sources, mu calpain is preferentially activated at the plasma membrane as evidenced by autolytic generation of faster migrating isoforms. By contrast, similar autolytic profiles for mu calpain in membrane or cytosolic fractions following addition of calcium were observed under cell-free conditions and within cells following death due to extended ionophore mediated calcium influx. These differential activation profiles for cytosolic mu calpain within living cells and following cellular fractionation/cell death indicate the presence of a regulatory system within neuronal cells. As in previous studies in other systems, we demonstrate the presence of a calpain activator protein. Cycloheximide treatment depleted the autolytic capacity of membrane-associated mu calpain within 4-6 hr without a corresponding decline in total mu calpain protein levels, indicating that the activator protein undergoes rapid turnover in comparison to calpain; pulse-chase radiolabeling confirmed the half-life of mu calpain to exceed 24 hr. Our data suggest that this labile protein represents a major rate-limiting step for in situ calpain activation within neuronal cells, and that, given the tremendous latent mu calpain activity within the cytosol, the interplay of the activator protein and the endogenous inhibitor calpastatin are crucial for maintaining neuronal homeostasis. PMID- 9210525 TI - Expression of DA11, a neuronal-injury-induced fatty acid binding protein, coincides with axon growth and neuronal differentiation during central nervous system development. AB - DA11 is the first fatty acid binding protein (FABP) for which gene expression has been shown to be upregulated following neuronal injury in the adult peripheral nervous system. To understand better the potential regulatory role(s) of this unique FABP in axonal growth and neuronal differentiation, we undertook a temporal and spatial study of DA11 gene expression in the developing rat central nervous system (CNS). Transient upregulation of DA11 mRNA and protein levels in CNS tissues were quantified by Northern blot hybridization and Western immunoblot analyses at different developmental ages. Homogenates of embryonic and neonatal cerebral cortex, cerebellum, brain stem, and hippocampal tissues contained 100 fold more DA11 mRNA and protein than corresponding adult tissues. Significant increase in DA11 mRNA was observed as early as embryonic day (E) 14 in cerebral cortex and cerebellum and E19 in brain stem and hippocampus. Postnatal levels of DA11 remained elevated through postnatal day (P) 10 in cerebral cortex, P14 in brain stem and hippocampus, and P20 in cerebellum. Localization of DA11-like immunoreactivity to specific CNS tissues, cell types, and intracellular compartments at P9 revealed a spatial pattern of neuronal expression different than that reported for other FABPs. DA11 protein was detected in the nucleus, cytoplasm, axons, and dendrites of differentiating neurons in cerebral cortex, hippocampus, cerebellum, brain stem, spinal cord, and olfactory bulb. The strong association of DA11 gene expression with development throughout the CNS suggests that this unique FABP plays an important role in axonal growth and neuronal differentiation in many different neuronal populations. PMID- 9210526 TI - Increased surface phosphatidylserine is an early marker of neuronal apoptosis. AB - Neuronal apoptosis is the subject of intense investigation and is beginning to be understood in some molecular detail. In the present study, we show that PC12 cells, like certain other cell types, redistribute phosphatidylserine (PS) from the inner leaflet to the outer leaflet of the plasma membrane early in the process of apoptosis. The externalised PS can be readily visualised by incubating intact cells with a fluorescent derivative of the protein annexin V. When apoptosis is blocked with an inhibitor of interleukin-1beta-converting-enzyme like proteases, the increased annexin binding is also blocked. Fluorescent annexin V binding provides a rapid and convenient way to identify apoptotic neurones. PMID- 9210527 TI - BAPTA-AM and ethanol protect cerebellar granule neurons from the destructive effect of the weaver gene. AB - The mechanisms by which the weaver gene (Reeves et al., 1989; Patil et al., 1995) inhibits neurite extension and/or induces death of the granule neurons in homozygous weaver mouse cerebellum are not presently understood. Here we show that BAPTA-AM and ethanol, which either reduce cytosolic levels of free calcium or prevent calcium entry, promote neurite outgrowth of the weaver neurons similar to the L-type calcium channel blocker verapamil (Liesi and Wright, 1996). Importantly, BAPTA-AM, ethanol, and verapamil not only restore neurite outgrowth of the weaver neurons but adjust their depolarized resting membrane potentials to the levels of normal neurons. These results indicate that calcium-dependent mechanisms mediate the action of the weaver gene and that the weaver neurons can be normalized by blocking this calcium effect. We further report that BAPTA-AM and verapamil also have a neuroprotective effect on normal neurons exposed to high concentrations of ethanol. We suggest that verapamil should be evaluated as a drug for treatment of alcohol-induced brain damage and neurodegenerative disorders. PMID- 9210528 TI - Age-associated alterations in hippocampal and basal forebrain nuclear factor kappa B activity. AB - Age-related cognitive deficits are often associated with loss of cholinergic activity within the neurotrophin-dependent cholinergic neurons that project from the basal forebrain to the hippocampus. The cause of reduced cholinergic function is unknown, but alterations in transcription factor-signaling pathways causing altered gene expression may cause decreased specific tissue function, resulting in loss of cholinergic activity. We measured transcription factor Nuclear Factor kappa B by electrophoretic mobility shift assay and Western analysis in young and aged rat brain tissues and report that basal levels of Nuclear Factor kappa B DNA binding activity increase in the hippocampus and basal forebrain with age to significantly higher levels at 30 months of age. This age-associated increase in binding activity is associated with increased translocation of p65 to the nucleus. These data show an age-associated alteration in Nuclear Factor kappa B signal transduction pathways that may contribute to age-associated decreases in specific tissue function. PMID- 9210529 TI - Platelet-derived growth factor delays oligodendrocyte differentiation and axonal myelination in vivo in the anterior medullary velum of the developing rat. AB - The AA dimeric form of platelet-derived growth factor (PDGF-AA) is implicated in the differentiation of cells of the oligodendrocyte lineage, which express PDGF receptors of the alpha subunit type (PDGF-alphaR). In the present study, we show that a single injection of PDGF-AA into the cerebrospinal fluid of neonatal rats delays oligodendrocyte differentiation and interrupts the progress of myelination in the anterior medullary velum (AMV), a white matter tract roofing the IVth ventricle of the brain. PDGF-AA or saline was injected intrathecally in postnatal day (P) 7 rats, and the AMV was subsequently removed and immunolabelled with the oligodendrocyte-specific antibody Rip, at P9, P12, and P21, corresponding to postinjection days (PID) 2, 5, and 14. At P9 (PID2), myelination was retarded in PDGF-AA-treated rats as opposed to saline-treated controls but progressed rapidly after P12 (PID5). Quantification supported the qualitative observations that PDGF AA mediated an acute decrease in the number of Rip+ oligodendrocytes at P9-12, which largely recovered by P21, suggesting that PDGF-AA may have delayed recruitment of myelinating oligodendrocytes. However, the definitive number of Rip+ oligodendrocytes in the AMV was not increased, suggesting that its action as a promoter of early oligodendrocyte survival may not ultimately affect the definitive number of myelinating oliogdendrocytes in vivo. We discuss the possibilities that excess PDGF-AA may have acted on early oligodendrocytes (precursors or preoligodendrocytes) to either (1) delay their differentiation by maintaining them in the cell cycle or (2) accelerate their differentiation, which may result in premature cell death in the absence of synchronised survival signals. This study supports a role for PDGF-AA in the timing of oligodendrocyte differentiation in vivo, as has been shown in vitro. PMID- 9210531 TI - Elevated intra-abdominal pressure increases plasma renin activity and aldosterone levels. AB - OBJECTIVE: To study the effects of elevated intra-abdominal pressure upon renal function and the renin-angiotensin-aldosterone system. MATERIALS AND METHODS: Two groups of anesthetized, ventilated swine were studied. Intra-abdominal pressure was increased in experimental animals (n = 6) by incrementally instilling an isosmotic ethylene glycol solution into the peritoneal cavity until intra abdominal pressure was 25 mm Hg above baseline. The intravascular volume was then expanded until cardiac index returned to baseline. Lastly, the solution was drained to decompress the abdomen. Control animals underwent surgical preparation but did not have their intra-abdominal pressure raised. Changes in systemic and pulmonary hemodynamic parameters, renal venous pressure, and urine output were recorded. Venous samples for plasma renin activity, aldosterone, and atrial natriuretic factor were drawn after each change in either intra-abdominal pressure or intravascular volume in experimental animals, and at the same time points in control animals. MEASUREMENTS AND MAIN RESULTS: Elevated intra abdominal pressure significantly (p < 0.05, analysis of variance) increased renal venous pressure, pleural pressure, wedge pressure, and pulmonary artery pressure compared to both baseline and control animals; whereas cardiac index and urine output decreased significantly. Both plasma renin and aldosterone levels increased significantly compared with baseline and controls. Intravascular volume expansion significantly increased urine output and decreased significantly both plasma renin activity and aldosterone levels. Abdominal decompression further significantly decreased both plasma renin activity and aldosterone levels. There were no significant changes in atrial natriuretic factor at any time point. CONCLUSIONS: Elevated intra-abdominal pressure decreases urine output and significantly up-regulates the hormonal output of the renin-angiotensin aldosterone system. Intravascular volume expansion in combination with abdominal decompression reverses the effects of acutely elevated intra-abdominal pressure upon renal function and the renin-angiotensin-aldosterone system. PMID- 9210530 TI - Reprioritization of liver protein synthesis resulting from recombinant human growth hormone supplementation in parenterally fed trauma patients: the effect of growth hormone on the acute-phase response. AB - BACKGROUND: One of the major components of the metabolic response to severe trauma is the alteration in concentrations of a large number of plasma proteins referred to as acute-phase proteins (APP). The principle mediators of these liver synthesized APP are mainly the cytokines interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha). METHODS: We have measured the plasma levels of IL-6, TNF alpha, and 20 APP in 24 adult, severely injured, hypermetabolic and highly catabolic patients with multiple injuries within 48-60 hours after injury, when they were receiving maintenance fluids without calories or nitrogen, and subsequently during 7 days of total parenteral nutrition with (n = 12) or without (n = 12) recombinant human growth hormone supplementation (rhGH, 0.15 mg/kg/d). RESULTS: Baseline positive APP due to severe trauma include C-reactive protein (CRP), alpha-1 antichymotrypsin, alpha-1 acid glycoprotein, alpha-1 antitrypsin, fibronectin, and factor B. Negative APP include IgG, IgM, complement-3, prealbumin, transferrin, ceruloplasmin, and albumin. Except for CRP, alpha-1 antichymotrypsin, and albumin, all the APP levels increase during 7 days of nutritional support. Plasma levels of cytokines IL-6 and TNF-alpha, although initially markedly increased after injury, decrease with parenteral refeeding. There is a linear correlation between CRP and IL-6 levels and also between the transport proteins prealbumin and transferrin. Trauma-induced increases in CRP and IL-6 levels decreased with nutrition alone, but did not change with rhGH supplementation. An immunosuppressed state of injury is evident from the decreased immunoglobulin levels (IgG, IgM, IgA) in the trauma patients. Total parenteral nutrition alone increases the immunoglobulin levels to normal. However, with adjuvant rhGH, only IgA levels are normalized. CONCLUSIONS: Adjuvant rhGH therapy does not attenuate the reprioritization of acute liver protein synthesis and results in only limited restoration of host defenses. The clinical implications of these findings await further study. PMID- 9210532 TI - Incidence of bacteremia after burn wound manipulation in the early postburn period. AB - BACKGROUND: Transient bacteremia associated with burn wound manipulation is considered a frequent occurrence and is commonly cited as an indication for perioperative antibiotic prophylaxis in burn patients. METHODS: In a prospective clinical setting, blood cultures (BC) were obtained from 19 burn patients at the following intervals: 30 minutes before wound cleansing (WC) or wound excision (WE), 30 minutes after the start of WC or WE, hourly until procedure completion, and 1 hour after completion. Burn wound biopsy for histologic grading and microbial culture was performed after the first BC. RESULTS: Twenty-two WC and 20 WE episodes were evaluated by 67 and 76 BC sets, respectively. Patients had a mean age of 42.8 years and mean burn size of 50% of the body surface area. Three WC episodes (13.6%) and four WE procedures (20.0%) were associated with postprocedure bacteremia. Two patients had both preprocedure and postprocedure bacteremia later attributed to nonburn wound infections. Excluding these cases, the bacteremia rate was 12.5% (9.5% from WC and 15% from WE). Wound biopsy culture and histologic analysis did not predict the occurrence of bacteremia. CONCLUSION: Current therapy is associated with a lesser incidence of burn wound manipulation-induced bacteremia than reported in prior series. The discordance between wound biopsy and BC results, the absence of positive histology, and the similarity of bacteremia occurrence rates with WC and WE confirm the effectiveness of current techniques of microbial control in burn wounds and question the need for perioperative antibiotic therapy in patients with burns involving less than 40% of the total body surface during the first 10 postburn days. PMID- 9210533 TI - Hospital readmission after trauma: an analysis of outpatient complications. AB - BACKGROUND: Outpatient complications leading to hospital readmission after hospitalization for trauma have not been examined. METHODS: A retrospective chart review of all trauma victims admitted to a Level 1 trauma center from January of 1990 to January of 1995 was performed to characterize patients who required readmission after hospitalization for trauma. Risk factors for readmission were determined by stepwise regression analysis. RESULTS: Of 15,463 trauma admissions, 209 patients (1.4%) required readmission, 84% within 30 days, 71% within 14 days. Reasons for readmission included wound (29%), abdominal (29%), pulmonary (18%), and thromboembolic (19%) complications. Fifty of the patients (24%) readmitted with a complication required an operation. Risk factors for readmission included: operation during first hospitalization (p < 0.0001), penetrating injury (p = 0.0001), and advanced age (p = 0.0001). Injury Severity Score, length of hospitalization, and gender were not independent predictors of readmission. CONCLUSIONS: Outpatient complications leading to readmission after hospitalization for trauma are not common; however, many are serious and require operative intervention. Because most complications were identified by the second week after discharge, outpatient follow-up visits should be scheduled within 7 to 14 days. Based on our findings, we recommend protocols be established to ensure follow-up for trauma patients, especially those who have had an operation, were victims of penetrating injury, or those > 65 years of age. PMID- 9210534 TI - Trauma patient outcome after the Prehospital Trauma Life Support program. AB - BACKGROUND: We have previously demonstrated a significant improvement in trauma patient outcome after the Advanced Trauma Life Support (ATLS) program in Trinidad and Tobago. In January of 1992, a Prehospital Trauma Life Support (PHTLS) program was also instituted. This study assessed trauma patient outcome after the PHTLS program. METHODS: Morbidity (length of stay and degree of disability), mortality, injury severity score, mechanism of injury, age, and sex among all adult trauma patients transported by ambulance to the major trauma hospital were assessed between July of 1990 to December of 1991 (pre-PHTLS, n = 332) and January of 1994 to June of 1995 (post-PHTLS, n = 350). RESULTS: Age, sex distribution, percentage blunt injury, and injury severity score were similar for both groups. Mortality pre-PHTLS (15.7%) was greater than post-PHTLS (10.6%). Length of stay and disability were statistically significantly decreased post-PHTLS. Age, injury severity score, and mechanism of injury were positively correlated with mortality in both periods. The previously reported post-ATLS mortality was similar to the pre-PHTLS mortality. CONCLUSIONS: Post-PHTLS mortality and morbidity were significantly decreased, suggesting a positive impact of the PHTLS program on trauma patient outcome. PMID- 9210535 TI - Hemodynamic effects of aortic occlusion during hemorrhagic shock and cardiac arrest. AB - OBJECTIVE: To determine the hemodynamic consequences of aortic occlusion during controlled hemorrhagic arrest. METHODS: Ten anesthetized, hemodynamically monitored swine were subjected to a 40 mL/kg hemorrhage over 10 minutes, followed by a 5-minute period of apnea. At this time (T15), they were randomized into an UP group (n = 5) in which the thoracic aorta was occluded or a DOWN group (n = 5) in which the aorta was not occluded. Simultaneously, volume resuscitation with shed blood plus 20 mL/kg of normal saline was performed over a 10-minute period. Cardiac massage was performed until return of spontaneous circulation (ROSC), which was defined as a sustained systolic blood pressure > 60 mm Hg. After 30 minutes of occlusion (T45), the aortic occlusion was released. Parameters measured include mixed venous and arterial blood gases, serum lactic acid levels, cardiac index, mean arterial pressure (MAP), mean pulmonary artery pressure (MPAP), coronary perfusion pressure (CoPP), and left ventricular stroke work index (LVSWI). Oxygen delivery index (DO2I) was measured using a pulmonary artery catheter, and oxygen consumption index (VO2I) was measured by direct calorimetry (Delta Trac metabolic monitor). RESULTS: Four animals in each group achieved ROSC after 3.0 +/- 1.8 and 2.2 +/- 1.8 minutes in the occluded and nonoccluded groups, respectively. During cardiac compressions and volume resuscitation, the CoPP, MAP, and MPAP were greater in the UP group, although the differences did not achieve statistical significance. After volume resuscitation was complete and during the period of aortic occlusion (T25-T45), the UP group had significantly greater MAP (mm Hg), with a difference of 42.5 +/- 20.75 mm Hg at T25 and 44.7 +/ 19 mm Hg at T35 (p < 0.03). Despite no difference in DO2I, VO2I (mL/min/kg) was significantly lower in the UP group than in the DOWN group, 4.28 +/- 0.48 versus 8.33 +/- 0.85 at T25 (p = 0.0002) and 4.62 +/- 0.9 versus 7.09 +/- 0.72 at T35 (p = 0.0005). After release of aortic occlusion at T45, the UP group had significantly lower CoPP (mm Hg) than the DOWN group (20.5 +/- 17.3 versus 66.5 +/- 28.2 at T45, p = 0.03). LVSWI (g/kg) was also lower in the UP than in the DOWN group (18.6 +/- 8.28 versus 36.5 +/- 10.2 at T60 [p = 0.031 and 23.6 +/- 6.48 versus 48.8 +/- 15.3 at T240 [p = 0.021). After release of the occlusion, there were trends toward increased acidosis and lactic acid levels in the UP group. CONCLUSIONS: Aortic occlusion in this controlled hemorrhagic arrest model does not result in improved salvage but is associated with impaired left ventricular function, systemic oxygen utilization, and coronary perfusion pressure in the postresuscitation period. PMID- 9210536 TI - Risk of hemorrhage and appropriate use of blood transfusions in pediatric blunt splenic injuries. AB - OBJECTIVE: To define changes in hematocrit (Hct) and the indications for blood transfusion in pediatric blunt splenic injury. DESIGN: Retrospective case series MATERIALS AND METHODS: All children with blunt splenic injuries from 1990 to 1995 were studied (n = 74). Transfusion practices were reviewed for the whole group. Thirty children with isolated splenic injuries who were not transfused were analyzed to document Hct changes (mean +/- 95% confidence intervals). MEASUREMENTS AND RESULTS: The Hct at presentation was 37 +/- 2%, which rapidly dropped to 31 +/- 2% (p < 0.05) within 24 hours. After remaining stable at that level for the next 5 days, the Hct rose to 33 +/- 4% on day 6 (p = not significant), 35 +/- 4% on day 7 (p = not significant), and 38 +/- 2% (p = not significant) on day 13 +/- 3. Fifteen children received transfusions, all but one of whom had suffered multiple injuries. The transfusion rate declined from 38% of children in 1990 to 10% in 1995. CONCLUSIONS: After the initial drop within the first 24 hours, the Hct remains stable and rises with time to reach the baseline by day 6. Transfusion rates have declined over time, and transfusions are now used almost exclusively in severely injured children with multiple injuries. PMID- 9210537 TI - Iliac vessel injury: operative physiology related to outcome. AB - BACKGROUND: Fifty-three patients treated at a level I trauma center with iliac vessel injury were studied to determine if body temperature and acid-base status in the operating room predicts outcome. METHODS: Records were reviewed for demographics, mechanism of injury, body temperature, acid-base status, operative management, and outcome. Statistical methods included Student's t test, odds ratio determination, and chi-square analysis to determine statistical significance. RESULTS: Fifty-three patients (47 male, 6 female) sustained 92 iliac vascular injuries (36 arterial, 56 venous). Mortality was 34%, with 72% of deaths due to shock within 24 hours. Physiologic parameters differed significantly between survivors and nonsurvivors. Odds ratio identified six conditions; the number present predicted outcome. CONCLUSIONS: (1) There are significant differences between initial and final operating room temperature and acid-base status in survivors versus nonsurvivors with iliac vessel injury. Conditions for odds ratio can be calculated and correlated with outcome. (2) A patient with two or more conditions should be considered for an abbreviated laparotomy to allow for reversal of "physiologic failure." PMID- 9210538 TI - The effect of a femoral fracture on concomitant closed head injury in patients with multiple injuries. AB - OBJECTIVE: We sought to determine the effect of a femoral shaft fracture, and its treatment by early intramedullary nailing, on the neurologic outcome of patients with multiple injuries with a concomitant head injury. DESIGN: Retrospective, case-control design using a prospectively gathered trauma data base. MATERIALS AND METHODS: We identified 46 patients with multiple injuries (mean Injury Severity Score [ISS] = 33.2) with closed head injuries (mean Glasgow Coma Scale [GCS] score = 7.8) and femur fractures, and matched as controls 99 patients with multiple injuries with head injuries but without femur fractures for age, sex, mechanism of injury, ISS (mean ISS = 34.0), and GCS (mean GCS score = 8.0). Follow-up parameters examined included early mortality, length of hospital or intensive-care unit stay, neuropsychological testing, and level of neurologic disability. RESULTS: There were no significant differences in the demographics or injury parameters between the study and control groups. There were no significant differences between the two groups in terms of early mortality (study group, 28%; control group, 27%; p = not significant), length of hospital/intensive-care unit stay (study group, 17.5/6.9 days; control group, 18.0/6.3 days; p = not significant), level of neurologic disability, or results of cognitive testing. CONCLUSION: Our study suggests that a femoral fracture in a patient with a concomitant head injury does not increase mortality or neurologic disability, and supports the continued early intramedullary nailing of femoral fractures for these patients. PMID- 9210539 TI - Diagnosis and management of closed internal degloving injuries associated with pelvic and acetabular fractures: the Morel-Lavallee lesion. AB - Closed internal degloving is a significant soft-tissue injury associated with a pelvic trauma in which the subcutaneous tissue is torn away from the underlying fascia, creating a cavity filled with hematoma and liquefied fat. It commonly occurs over the greater trochanter but may also occur in the flank and lumbodorsal region. When this closed internal degloving occurs over the greater trochanter, it is known as a Morel-Lavallee lesion. We reviewed 24 patients who sustained a closed internal degloving injury. Cultures from the closed internal degloving injury were positive in 46% (11 of 24 cases). The incidence of positive cultures was not dependent on the time from injury to debridement. All wounds were treated by thorough debridement before or during pelvic or acetabular surgery. Three patients subsequently developed deep-bone infections, only one of whom had a positive culture at the initial debridement. One patient whose wound was primarily closed over suction drains developed a chronic deep soft-tissue infection requiring multiple debridements. The development of hematoma in the zone of operation reduces the safety of early operative intervention by increasing the risk of infection. An expanding hematoma in a closed internal degloving injury may further compromise the skin vascularity if not promptly drained. The injured soft tissues should be debrided early, either before or at the time of fracture fixation. The wound should be left open, and repeated surgical debridement of the injured tissue is recommended. PMID- 9210540 TI - Open reduction and internal fixation of combined fourth and fifth carpometacarpal (fracture) dislocations. AB - BACKGROUND: Traditionally, combined fourth and fifth carpometacarpal fracture dislocations are treated conservatively or by means of Kirschner wire after closed reduction. Since 1983, unstable dislocations have been treated with open reduction and screw fixation or with a temporary plate that bridges the fourth carpometacarpal (CMC) joint to maintain anatomical reduction. METHODS: In a retrospective study, we evaluated the results of this surgical approach in a group of 11 patients and another group of 4 conservatively treated patients. RESULTS: Eleven patients were treated by means of open reduction and rigid screw fixation (n = 6) or plate bridging of the fourth CMC joint (n = 5). Reduction and fixation of the fourth CMC joint always led to spontaneous anatomical reduction of the fifth CMC joint. At long-term follow-up, nine of these patients had full recovery of their hand function without any complaints. CONCLUSION: Open reduction and internal fixation of unstable ulnar CMC dislocations produced excellent results. PMID- 9210541 TI - Prediction of posttraumatic adult respiratory distress syndrome by albumin excretion rate eight hours after admission. AB - BACKGROUND: Adult respiratory distress syndrome (ARDS) in trauma victims carries a mortality on the order of 50%. An early feature is an increased capillary permeability causing an extravasation of plasma proteins and water, leading to interstitial edema. In the kidney, the increase in microvascular permeability is manifested as increased albumin excretion detectable by sensitive immunoassay. METHODS: Forty seven trauma victims were studied for 5 days; 32 of them had Injury Severity Scores > 18. A diagnosis of ARDS was made on the recommendations of the American-European Consensus Conference on ARDS (1994). Eight patients developed ARDS, five developed pulmonary dysfunction, and the remainder showed no significant pulmonary abnormality. RESULTS: Using the near patient urine albumin immunoassay, albumin excretion rate (AER) was measured after admission. For patients with Injury Severity Score > 18, the median (95% confidence interval) AER 8 hours after admission was 63 (range, 40-99) microg per minute for those without impaired lung function and 339 (range, 162-454) microg per minute for those in the combined ARDS and pulmonary dysfunction group (Mann-Whitney test, p = 0.0004). The median AER was 51 (range, 27-98) microg per minute for patients with Injury Severity Score < 18. The positive predictive value for the development of ARDS or pulmonary dysfunction of AER > 130 microg per minute was 85%, with a negative predictive value of 95%. CONCLUSIONS: These data indicate that the capillary leak associated with the subsequent development of pulmonary dysfunction and ARDS can be detected within 8 hours of admission at the patient's bedside, thus providing a means of early identification of patients at greatest risk and allowing for early intervention. PMID- 9210542 TI - Measurement of right ventricular performance during apnea in patients with acute lung injury. AB - BACKGROUND: Mechanical ventilation resulted in increased, decreased or unchanged end-diastolic volumes together with either profoundly decreased or unchanged right ventricular ejection fraction (RVEF). The goal of our study was, therefore, to evaluate the effects of positive end-expiratory pressure on measurements of RVEF performed during apneic periods with different levels of positive end expiratory pressure. METHODS: Fifteen consecutive patients suffering from acute lung injury after major surgery or trauma were included. Measurements were performed during 15 seconds of apnea at airway pressure levels of 0 (baseline), 10, 20, and 30 cm H2O. Cardiac output and RVEF were determined using the thermodilution technique. RESULTS: Lung inflation to an airway pressure of 30 cm H2O caused a 22 +/- 14% decrease of cardiac output resulting from a 20 +/- 14% decrease of stroke volume index. The decrease of stroke volume index was induced by a 17 +/- 11% decrease of right ventricular end-diastolic volume index, while RVEF remained virtually unchanged (0.49 +/- 0.10 vs. 0.47 +/- 0.12 at 0 and 30 cm H2O, respectively). Relative changes of cardiac output were closely correlated with changes of right ventricular end-diastolic volume index (p < 0.05, r2 = 0.78). CONCLUSIONS: Right ventricular systolic function was well maintained despite substantially decreased end-diastolic volumes. In our study, during apneic conditions, higher levels of positive end-expiratory pressure did not worsen RVEF in patients with acute lung injury. The proposed technique of apneic lung inflation may serve as an alternative approach to obtain comparable measurements of RV function in patients with acute lung injury. PMID- 9210543 TI - Increased neutrophil elastase, persistent intravascular coagulation, and decreased fibrinolytic activity in patients with posttraumatic acute respiratory distress syndrome. AB - BACKGROUND: To investigate the role of plasma neutrophil elastase (elastase alpha1-proteinase inhibitor complex), plasminogen activator inhibitor-1 (PAI-1), and disseminated intravascular coagulation (DIC) in patients with posttraumatic acute respiratory distress syndrome (ARDS) and to explore the time course of the changes of these factors after trauma, we performed a prospective case-control study. METHODS: The study subjects consisted of 41 trauma patients, 5 with ARDS, 7 at risk for but not developing the syndrome, and 29 control patients without or with no risk for ARDS. Plasma neutrophil elastase, PAI-1 activity, and PAI-1 antigen concentration were measured on the day of the injury and on days 1, 3, and 5 after admission. DIC was measured on the basis of the DIC score. The results of these measurements and demographic data were compared among the three groups. RESULTS: Neutrophil elastase, PAI-1 activity, and PAI-1 antigen concentration for the ARDS patients continued to be markedly high until the fifth day of admission, and the values on the fifth day were significantly higher than those of the other two groups. All patients with ARDS developed DIC. A decrease in the DIC score was found for the control patients and also for the patients at risk for ARDS; however, for the patients with ARDS, the DIC score did not improve during the study period (p = 0.5809). CONCLUSION: We provide precise information on the time course of neutrophil elastase, PAI-1, and DIC in trauma patients with ARDS and those at risk of developing this syndrome. Neutrophil activation and persistent intravascular coagulation as well as impaired fibrinolysis may play a role in the pathogenesis of posttraumatic ARDS. PMID- 9210544 TI - Influence of selective decontamination of the digestive tract on cell-mediated immune function and bacteria/endotoxin translocation in thermally injured rats. AB - OBJECTIVE: To determine the influence of pretreatment with selective decontamination of the digestive tract (SDD) on systemic immunosuppression, and the relationship between bacteria/endotoxin translocation and abnormalities of immune function in thermally injured rats. DESIGN, MATERIALS, AND METHODS: Animals were subjected to a 40% full-thickness scald injury, and divided into SDD treated and control groups. The treatment group received SDD (polymyxin E, tobramycin, and 5-flucytosine) by gavage twice daily for 3 days before the experiment and continued for 5 days after thermal injury. The control group was given the same amount of water. The parameters reflecting cell-mediated immunity, including splenocyte proliferation in response to mitogens, interleukin 2 (IL-2) production, and lymphocyte subpopulation, were measured before injury and 1 and 5 days after burn, respectively. MEASUREMENTS AND MAIN RESULTS: Thermal injury resulted in marked reduction in splenocyte proliferative response to T-cell mitogens, IL-2 production, and T-helper/suppressor cells (CD4/CD8) ratio. Prophylactic treatment with SDD significantly decreased the incidences of bacterial translocation and endotoxemia, prevented suppressive mitogenic response and inadequate IL-2 production (p < 0.05-0.01) but did not affect the abnormal ratio of CD4 to CD8 T lymphocytes in blood (p > 0.05). CONCLUSIONS: These results suggest that bacteria/endotoxin translocation from the gut appears to be involved in cell-mediated immune dysfunction as a consequence of thermal injury. Pretreatment with SDD might attenuate postburn immunosuppression by preventing translocation events. PMID- 9210545 TI - Basic fibroblast growth factor reduces the gut and liver morphologic and functional injuries after ischemia and reperfusion. AB - OBJECTIVE: To explore the possible effects of basic fibroblast growth factor (bFGF) on ischemic gut and liver injuries after trauma. METHODS: Animal models of superior mesenteric artery occlusion (45 minutes) and reperfusion (3 days) were used in this study. Seventy-two Wistar rats were divided into three groups of 24 rats each. The animals in bFGF-treated group were injected with 4 microg bFGF/rat in 0.15 mL normal saline solution containing heparin 0.1% (w/v) through the jugular vein at the onset of reperfusion. In the normal saline control group, all rats received the same vehicle, but without bFGF. Group 3 (sham-operated) underwent the same laparotomy procedure, but without superior mesenteric artery occlusion. Liver function parameters, the levels of serum tumor necrosis factor alpha, nitric oxide, superoxide dismutase, malondialdehyde (MDA), tissue bacterial examination, and pathologic study were used to evaluate the results. RESULTS: In bFGF-treated rats, the amounts of serum alanine transaminase and aspartate aminotransferase and serum tumor necrosis factor-alpha were reduced significantly at 6, 24, and 48 hours when compared with normal saline-treated rats. However, the changes in nitric oxide, superoxide dismutase, and MDA varied from each other as a function of time after injury. The amounts of nitric oxide were increased significantly at 6 hours in intestine in normal saline-treated rats and in liver in bFGF-treated rats (p < 0.05). At 6 hours after reperfusion, the activity of superoxide dismutase in normal saline-treated rats were much lower in liver than those in bFGF-treated and sham-operated rats (p < 0.05), but the levels of MDA were increased in intestine in bFGF-treated rats and in liver in normal saline-treated rats when compared with sham-operated rats (p < 0.05). At 24 hours, the levels of MDA in normal saline-treated rats were much higher than those in both bFGF and sham-operated rats (p < 0.05). Bacterial examination revealed that the ratio and the amounts of bacterial translocation from gut to liver, spleen, and mesenteric lymph nodes in bFGF-treated rats were much lower than those in normal saline-treated rats. The results of pathologic study support the assumption that bFGF provided protective effects against reperfusion injury. CONCLUSIONS: Intravenous administration of bFGF may benefit in reducing gut and liver injuries after ischemia and reperfusion. The mechanisms of those effects may involve mitogenic and nonmitogenic effects of bFGF. PMID- 9210546 TI - Ultrasound based key clinical pathway reduces the use of hospital resources for the evaluation of blunt abdominal trauma. AB - BACKGROUND: Evaluating blunt abdominal trauma remains a resource intensive aspect of trauma care. Recently, emergency department ultrasound has been promulgated as a noninvasive diagnostic alternative. Consequently, we hypothesized that an ultrasound based key clinical pathway (KCP) would reduce the number of diagnostic peritoneal lavage (DPL) and computed tomographic (CT) scans required to evaluate blunt abdominal trauma without increased risk to the patient. METHODS: This study was a prospective analysis of patients evaluated for blunt abdominal trauma during a 3-month period using this KCP compared with a 3-month historical cohort. RESULTS: Data were collected for 486 KCP patients and were compared with 516 patients in the study cohort. No differences were noted regarding demographics, number of laparotomies, or type of injuries. Using the KCP, DPL was reduced from 17 to 4%, and computed tomography from 56 to 26%. Furthermore, the injury severity score increased from 11.6 to 21.5 for DPL patients and from 4.6 to 8.3 for computed tomography patients. Ultrasound exams were used exclusively in 65% of patients. CONCLUSIONS: An ultrasound based KCP resulted in significant reductions in the use of invasive DPL and costly CT scanning in the evaluation of blunt abdominal trauma without risk to the patient. PMID- 9210547 TI - Treatment of liver injuries at level I and level II centers in a multi institutional metropolitan trauma system. The Midwest Trauma Society Liver Trauma Study Group. AB - OBJECTIVE: The development of trauma systems and trauma centers has had a major impact on the fate of the critically injured patient. However, some have suggested that care may be compromised if too many trauma centers are designated for a given area. As of 1987, the state of Missouri had designated six adult trauma centers, two Level I and four Level II, for the metropolitan Kansas City, Mo, area, serving a population of approximately 1 million people. To determine whether care was comparable between the Level I and II centers, we conducted a concurrent evaluation of the fate of patients with a sentinel injury, hepatic trauma, over a 6-year period (1987-1992) who were treated at these six trauma centers. METHODS: All patients during the 6-year study period who suffered liver trauma and who survived long enough to be evaluated by computerized tomography or celiotomy were entered into the study. Patients with central nervous system trauma were excluded from analysis. Information concerning mechanism of injury, RTS, Injury Severity Score (ISS), presence of shock, liver injury scoring, mode of treatment, mortality, and length of stay were recorded on abstract forms for analysis. Care was evaluated by mortality, time to the operating room (OR), and intensive care unit (ICU) and hospital length of stay. RESULTS: Over the 6-year period 300 patients with non-central nervous system liver trauma were seen. Level I centers cared for 195 patients and Level II centers cared for 105. There was no difference in mean ISS or ISS > 25 between Level I and II centers. Fifty-five (28%) patients arrived in shock at Level I centers and 24 (23%) at Level II centers. Forty-eight patients (16%) died. Thirty-two (16%) died at Level I centers, and 16 (15%) died at Level II centers. Twenty of 55 patients (36%) in shock died at Level I centers, and 11 of 24 (46%) died at Level II centers (p = 0.428). Forty-three patients (22%) had liver scaling scores of IV-VI at Level I centers, and 10 (10%) had similar scores at Level II centers (p < 0.01). With liver scores IV-VI, 22 of 43 (51%) died at Level I centers and 10 of 14 (71%) died at Level II centers (p = 0.184). There was no difference in mean time or in delays beyond 1 hour to the OR for those patients in shock between Level I and II centers. There was a longer ICU stay at Level II centers (5.0 +/- 8.3 vs. 2.8 +/- 8.4 days, p = 0.04). This difference was confined to penetrating injuries. There was no difference in hospital length of stay. CONCLUSIONS: In a metropolitan trauma system, when Level I and II centers were compared for their ability to care for victims of hepatic trauma, there was no discernible difference in care rendered with respect to severity of injury, mortality, delays to the OR, or hospital length of stay. It was observed that more severe liver injuries were seen at Level I centers, but this did not seem to significantly affect care at Level II centers. There was a longer ICU stay observed at Level II centers owing to penetrating injuries, possibly because there were fewer penetrating injuries treated at these facilities. Although the bulk of patients were seen at Level I centers, care throughout the system was equivalent. PMID- 9210548 TI - Conservative management of pancreatic trauma in children. AB - Many adults and most children with a solid-organ abdominal injury can be managed nonoperatively. To date, however, little is known about the outcome of nonoperative management of pancreatic injury. To analyze current treatment patterns of pancreatic injury in children, all children (age < 19 years) identified in the National Pediatric Trauma Registry (49,540 patients) and admitted to two level I pediatric trauma centers with a diagnosis of injury to the pancreas (International Classification of Disease-9 codes 863.81-863.84 and 863.91-863.94) were reviewed. Over a 7-year period, 154 children were identified with pancreatic injury. Thirty-one (20%) sustained severe injuries (grades III, IV, or V) and 123 (80%) sustained lower-grade injuries (grades I and II). Sixteen (52%) of the children sustaining grades III, IV, or V injury required pancreatic procedures (9 distal resections, 3 simple repairs, 2 enteric anastomoses, 2 others). Only 26 (21%) of the grades I and II injuries required surgical intervention specific to the pancreas (11 resections, 9 catheter drainage of pseudocysts, 2 enteric anastomoses, 4 others). Ninety-seven (79%) grades I and II injuries were successfully managed conservatively. Overall, 15 (10%) children required drainage procedures for pseudocyst. The frequency of operative intervention decreased during the last 4 years of the study (18 vs. 26%, p > 0.05), coinciding with a decrease in the frequency of drainage procedures for pseudocysts. The need for surgical intervention was not influenced by age, Injury Severity Score, or Pediatric Trauma Score (p > 0.05). Associated abdominal injuries were common but did not influence operations on the pancreas (p > 0.05). No deaths were attributed to the pancreatic injury. These data indicate that early intervention for pancreatic injury, in the absence of clinical deterioration or major ductal injury (grades III, IV, or V), is unwarranted, and careful observation may supplant the conventional surgical therapy recommended for adults. PMID- 9210549 TI - Production of tumor necrosis factor-alpha and interleukin-1beta by human cerebral microvascular endothelium after percussive trauma. AB - Intracerebral cytokine production is thought to be partially responsible for the brain edema and increased leukocyte adhesion seen after head injury by both a direct effect on vascular permeability and by causing leukocyte activation. Cerebrospinal fluid concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, and IL-6 are elevated after traumatic brain injury. The cerebral endothelium has not been investigated as a de novo source of cytokines after injury. We have found that conditioned media from cultured human cerebral microvascular endothelium (HCME) subjected to percussion trauma increases neutrophil chemotaxis. To test the hypothesis that percussive trauma increases the production of TNF-alpha and IL-1beta by HCME, serial supernatant samples from passage 2 HCME were collected for 24 hours and analyzed for TNF-alpha and IL 1beta concentration by enzyme-linked immunosorbent assay after trauma. HCME subjected to percussion injury secreted significantly more TNF-alpha at 8 and 24 hours and significantly more IL-1beta at 4 and 24 hours compared with uninjured controls (p < 0.05, Student's t test). These data suggest that HCME production of inflammatory cytokines occurs after traumatic brain injury independent of systemic influences. In situ cytokine production by HCME after percussion trauma may mediate the increased cerebral leukocyte accumulation and cerebrovascular dysfunction observed after focal brain injury. PMID- 9210550 TI - Outcome analysis of patients with severe head injuries and prolonged intracranial hypertension. AB - OBJECTIVE: To describe the functional outcome of a select group of patients with severe head injuries who would a priori be assumed to have a dismal outcome and to determine prognostic factors that can be used for effective family counseling and rational utilization of scarce resources. METHODS: Thirty-seven patients with severe head injuries (admission Glasgow Coma Scale (GCS) score < 8) with prolonged (> 96 hours) intracranial hypertension were studied. Parameters recorded included admission age, GCS, evidence of prehospital hypotension, initial computed tomography findings, intracranial pressure (ICP), cerebral perfusion pressure (CPP), and therapeutic intensity level. RESULTS: Thirty-eight percent of patients in this study achieved a Glasgow Outcome Scale score (GOS) of 4 (moderate disability) or better when assessed 1 year after injury. Patients who achieved these good outcomes were significantly younger (mean 23.6 +/- 8.8 years) than patients who were severely disabled or worse (GOS 1-3) (34.3 +/- 15.0 years) (p = 0.0098). The mean admission GCS in the good-outcome group tended to be higher than that of the poor-outcome group (5.8 +/- 1.5 vs 4.8 +/- 1.6, p = 0.065). When patients with good outcomes (GOS 4 or 5) were compared with those with poor outcomes (GOS 1-3), no significant differences in mean or peak ICP, percentage of time intervals with elevated ICP, lowest recorded CPP, or length of ICP monitoring were detected. CONCLUSION: Younger patients, particularly those with GCS > 5, have the potential for excellent recovery despite prolonged (> 96 hours) intracranial hypertension. These patients will benefit from continued aggressive ICP and CPP management. PMID- 9210551 TI - Infusion of hot crystalloid during operative burn wound debridement. AB - BACKGROUND: Hypothermia exacerbates coagulopathy and is thus a potentially devastating morbidity during operative debridement of burn wounds. Current techniques for maintaining body temperature include warming intravenous fluids at 38 degrees C. The purpose of this study was to assess the safety of infusing saline heated to 55-60 degrees C. METHODS: Using a modified fluid warmer, saline heated to 60 degrees C was infused through central venous access in eight adult patients undergoing debridement of burn wounds. The temperature of the saline actually entering the patient was measured by a thermocouple attached at the connection to the central line catheter. RESULTS: The actual infusate temperature was 54.0 +/- 1.2 degrees C. Over the first hour, 1,100 mL of hot saline was given, thus delivering 17.6 kcal more heat than fluid warmed to the traditional 38 degrees C. Core temperature measured via esophageal and Foley catheters had an insignificant trend toward increase during the operative procedure. There was no evidence of intravascular hemolysis or coagulopathy. CONCLUSION: This pilot study suggests that infusion of hot crystalloids given via central venous access is safe and may be an acceptable adjuvant in attenuating hypothermia during operative procedures. PMID- 9210552 TI - Medical costs and economic production losses due to injuries in the Netherlands. AB - BACKGROUND: To support injury control, we assessed the direct medical costs and indirect costs of injuries in the Netherlands, making use of recent advances in health economics. METHODS: We estimated the direct medical costs with the help of available data on health care utilization as a consequence of injuries. In our calculations of indirect costs, we used two alternative approaches. We used the traditional human-capital approach, which estimates the potential economic production losses caused by diseases or injuries. In addition, we applied the friction-costs method, which was recently developed as an attempt to measure the actual economic production losses to society. RESULTS: Injuries are an important source of medical costs and economic production losses. Almost two-thirds of the medical costs are the result of injuries among females (mainly domestic injuries of elderly women). On the contrary, independent of the method used, more than 80% of the indirect costs are the result of injuries among males (mainly caused by a high frequency of traffic injuries, occupational injuries, and sports injuries among young males). The application of the friction-costs method confirms the importance of injuries as a source of production losses in comparison with other diseases, showing that they belong to the main three causes of indirect costs to society. CONCLUSIONS: Estimates of the medical costs and both the potential and actual economic production losses to society clearly demonstrate that injuries should be a major concern for health policy makers and the medical profession. PMID- 9210554 TI - Modeling injury outcomes using time-to-event methods. AB - BACKGROUND: Mortality is an important measurement of injury outcomes, but measurements reflecting disability or cost are also important. Hospital length of stay (LOS) has been used as an outcome variable, but reduced LOS could be achieved either by improved care or by increased mortality. A solution to this statistical problem of "competing risks" would enable injury outcomes based on LOS to be modeled using time-to-event methods. METHODS: Time-to-event methodology was applied to 2,106 cases with complete data from the 1991-1994 registry of a regional trauma center. LOS was used as the outcome variable, modified by assigning an arbitrarily long LOS to any fatal case. A combination of proportional hazards and logistic regression models was used to explore the effects of potential predictive variables, including Trauma Score (TS), Injury Severity Score (ISS), components of TS or ISS, age, sex, alcohol use, and whether a patient was transferred. RESULTS: The "TRISS" combination of TS, ISS, and age previously shown to predict mortality also predicted "modified LOS" (Wald p value less than 0.001 for each variable). Models using only age and certain components of ISS or TS fit our data even better, with fewer parameters. Other variables were not predictive. Modified Kaplan-Meier plots provided easily interpreted graphical results, combining both mortality and LOS information. CONCLUSIONS: With a simple modification to allow for competing risks, time-to-event methods enable more informative modeling of injury outcomes than binary (lived/died) methods alone. Such models may be useful for describing and comparing groups of hospitalized trauma patients. PMID- 9210553 TI - The Colorado motorcycle safety survey: public attitudes and beliefs. AB - BACKGROUND: Motorcycle riders have a high risk of traumatic brain injury, disability, and death. Epidemiologic studies have proven that helmets reduce the severity of brain injuries and the cost of care. Yet, Colorado remains one of three states with no helmet law for riders. OBJECTIVES: This study measured public support for (1) a mandatory motorcycle helmet use law and (2) mandatory motorcycle operator safety training. We also sought to ascertain citizens' attitudes toward traffic safety mandates from the federal government. METHODS: Structured telephone interviews were conducted with 407 Colorado adults selected by random-digit dialing. RESULTS: Sixty-five percent of respondents believed that motorcycle riders of all ages should be required to wear helmets. An additional 18% believed that only riders under age 21 should be required to wear helmets. Only 17% of respondents opposed all helmet laws. Even among motorcyclists, most supported helmet laws for all riders (47%) or for those <21 years of age (26%). In a multiple logistic regression, there were three significant independent predictors of a pro-helmet law stance: older age, female gender, and not possessing a motorcycle operator's license. Most respondents also supported mandatory motorcycle operator safety training. Despite supporting state helmet use regulations, a large proportion (41%) opposed mandatory Federal mandates to enact them. CONCLUSION: Even in Colorado, a state with no helmet use requirements, there is strong public support for a regulatory strategy of motorcycle helmet use laws. PMID- 9210555 TI - Posterior circulation cerebral infarcts associated with repair of thoracic aortic disruption using partial left heart bypass. AB - BACKGROUND: Partial left heart bypass is widely used in the repair of traumatic aortic disruptions. We recently encountered two patients with posterior circulation infarctions after repair of traumatic aortic disruptions using heparin-less partial left heart bypass. METHODS/RESULTS: Both patients underwent interposition graft repair of thoracic aortic transections at the level of the isthmus. The first patient developed a left posterior inferior cerebellar artery infarct after a clamp time of 44 minutes. Swelling of this infarct necessitated ventriculostomy placement. The second patient developed a pontine infarct postoperatively after a cross-clamp time of 56 minutes and suffered a persistent left upper extremity paresis. CONCLUSIONS: Partial left heart bypass may have predisposed these two patients to clamp-related embolic events via the left vertebral artery. This experience warrants further surveillance to detect these infarcts which can require neurosurgical intervention. Additionally, the events suggest reconsideration of systemic anticoagulation during aortic cross-clamp times exceeding 30 minutes. PMID- 9210556 TI - Direct repair of a giant extracranial vertebral artery pseudoaneurysm through the aneurysmal cavity. PMID- 9210557 TI - Liver injuries and two-point shoulder restraints: case report and discussion. PMID- 9210558 TI - Airbag-induced lethal cervical trauma. AB - In a frontal collision of a car (taxicab) perpendicular into a streetcar with an impact speed of approximately 30 kph (20 mph), the driver survived with minor injuries. The front-seat passenger was extremely "out-of-position," with her seat positioned nearly fully forward. This in combination with her short stature led to fatal injuries resulting from the inflating airbag (U.S.-type) striking against her face and chin. At the scene, she was found essentially clinically dead, but was resuscitated and died finally 13 days later. Postmortem examination showed a complete disruption of all ventral ligaments between the base of the skull and the first and second vertebrae, a nearly complete ventral rupture of the medulla, and diffuse axonal injury of the brain. PMID- 9210560 TI - Mennen plate fixation for fractures of the femoral shaft after ipsilateral hip arthroplasty. AB - We performed osteosynthesis with the use of a Mennen plate for six patients with femoral fractures in the vicinity of the stem, which occurred after ipsilateral hip arthroplasty. The fixation was so favorable that postoperative deformity was slight. At 4 months after surgery, bone fusion was obtained in all patients, and their hip joint functions recovered to the preinjury level. For femoral fractures in the vicinity of the stem after ipsilateral arthroplasty, there are no suitable fixation methods other than treatment with the Mennen plate. Therefore, this method is recommended. PMID- 9210559 TI - Overdistraction of cervical spine injuries with the use of skull traction: a report of two cases. AB - Two cases in which cervical spine overdistraction occurred with the use of skull traction are described. A summary of the clinical presentations and definitive treatment together with some bibliographic references are discussed. Finally, suggestions regarding how to avoid overdistraction when using skull traction are given. PMID- 9210561 TI - Intracerebral venous thrombosis and hematoma secondary to high-voltage brain injury. AB - We report the case of a 19-year-old male who sustained an electrodynamic (16.67 Hz) high-voltage (15,000 V) railway overhead cable injury. He lost consciousness 30 minutes after contact and died secondary to brainstem herniation as a result of intracerebral swelling within 8 days. Repeated cranial computed tomography revealed a huge hemispheric mass bleeding accompanied by subarachnoidal hemorrhage. Additionally, necropsy showed an extensive thrombosis of the adjacent cerebral veins. The pathophysiological mechanism of this unusual injury is discussed. PMID- 9210562 TI - A simple stenting method for management of hepatic ductal injury secondary to blunt abdominal trauma: two case reports. AB - We report two cases of liver injury with hepatic ductal disruption after blunt abdominal trauma. The first case involves a 23-year-old male. Because the bifurcation of the hepatic duct was longitudinally torn, two stenting catheters were inserted toward the right and left hepatic ducts without suture closure of the tear. The patient is well 10 years after the injury. The second case involves a 22-year-old male who suffered an infarction of the inferior portion of the medial segment of the left hepatic lobe as well as a laceration of the left hepatic duct, a 50% circumferential tear. A stenting catheter was introduced into the left hepatic duct, but the defect was not sutured. The patient is well 1.5 years after the injury. The catheter stenting method without suture repair or defect plasty is a simple and effective way to manage hepatic ductal injury. PMID- 9210564 TI - Posterior dislocation of the shoulder associated with fracture of the humeral anatomic neck. PMID- 9210563 TI - Massive pelvis injuries treated with amputations: case reports and literature review. PMID- 9210565 TI - Munchausen syndrome presenting as trauma. AB - Rarely, a patient with Munchausen syndrome will present with apparent trauma. A computerized literature search from 1966 until the present discovered only three such case reports, none of which appeared in a surgical journal. We report a fourth case. The characteristics of Munchausen syndrome are illustrated. The possibility that such a patient may have a true injury is also discussed. PMID- 9210566 TI - Irreversible shock revisited: mechanical support of the cardiovascular system: a case report and review. PMID- 9210567 TI - Delayed presentation and rupture of a posttraumatic innominate artery aneurysm: case report and review of the literature. PMID- 9210568 TI - Impact of traumatic subarachnoid hemorrhage on outcome in nonpenetrating head injury. Part II: relationship to clinical course and outcome variables during acute hospitalization. PMID- 9210569 TI - Use of peroxidase substrate Vector VIP for multiple staining in light microscopy. AB - The study of the distribution of a fiber input to a particular brain area and the visualization of the anatomical relationships of that input with both projection- and interneurons, requires a triple-staining that allows the unequivocal distinction of each of the three components in one and the same histological section. In this regard, we investigated the properties of a recently introduced peroxidase chromogen, VIP (V-VIP; Vector Labs) in combination with two traditional substrates, standard diaminobenzidine (DAB, brown precipitate) and nickel-enhanced DAB (DAB-Ni, black). In rats, the anterograde tracer biotinylated dextran amine (BDA) and the retrograde tracer fluorogold (FG) were injected in the perirhinal cortex and hippocampus, respectively. Transported BDA was detected with an avidin-biotin-peroxidase complex, whereas the transported FG was detected via a PAP method. Tracing with BDA and FG was combined with parvalbumin- or calbindin-immunocytochemistry. We compared various combinations and staining sequences. The best results were obtained with a staining sequence comprising first the BDA stain with DAB-Ni as chromogen, second the FG protocol with the chromogen DAB and finally, parvalbumin- or calbinding-immunocytochemistry using the chromogen V-VIP. The order with which the chromogens were applied appeared to be critical. Partial or even total loss of V-VIP reaction product has been observed after standard dehydration in ethanol. As an alternative, a quick dehydration procedure in toluene yields much better staining. Colour separation is excellent and the sensitivity is high. This procedure may also be used for detection of any other combination of three different labels, taking the usual care to avoid cross-reactivity between antibodies. PMID- 9210570 TI - Persistent neuronal labeling by retrograde fluorescent tracers: a comparison between Fast Blue, Fluoro-Gold and various dextran conjugates. AB - The permanence of retrograde neuronal labeling by the fluorescent tracers Fast Blue, Fluoro-Gold, Mini-Ruby, Fluoro-Ruby and Fluoro-Emerald was investigated in adult rat spinal motorneurons at 1, 4, 12 and 24 weeks after tracer application to a transected muscle nerve. After 1 week, the largest number of retrogradely labeled motoneurons was found with Mini-Ruby, Fluoro-Gold and Fluoro-Ruby, while Fluoro-Emerald yielded a smaller number of labeled cells. With increasing survival time, all of these tracers exhibited a marked decrease in the number of labeled neurons. Fast Blue also produced very efficient staining after 1 week and, in addition, the number of Fast Blue-labeled cells remained constant over the entire time period studied. Also in embryonic spinal cord tissue exposed to Fast Blue. the label persisted for at least 6 months after transplantation into adult spinal cord. Double-labeling experiments combining Fast Blue with Fluoro Gold, Mini-Ruby, Fluoro-Ruby or Fluoro-Emerald showed that all these substances were non-toxic and that the time-related decrease in the number of neurons labeled by the latter tracers was due to degradation or leakage of the dyes. Thus, Fast Blue would be the tracer of choice for motoneuronal labeling in long term experiments, whereas the usage of the other tracers should be restricted to experiments of limited duration. PMID- 9210571 TI - Effective spread and timecourse of neural inactivation caused by lidocaine injection in monkey cerebral cortex. AB - We studied the effective spread of lidocaine to inactivate neural tissue in the frontal cortex of the rhesus monkey. Injections of 2% lidocaine at 4 microl/min were made while units were recorded 1 or 2 mm away. To inactivate units 1 mm away from the injection site 100% of the time, 7 microl of lidocaine had to be injected. To inactivate units 2 mm away from the injection site 100% of the time, 30 microl of lidocaine were required. Units were maximally inactivated around 8 min after the start of a lidocaine injection, and they gradually recovered, regaining most of their initial activity by around 30 min after the start of an injection. The volume of lidocaine required to inactivate neurons > 90% of the time could be estimated by the spherical volume equation, V = 4/3 pi (r)3. To prolong the inactivation, a slower infusion of lidocaine subsequent to an initial bolus was effective. Saline control injections had no effect. These results allow both a prediction of the timecourse of neural inactivation and an estimate of the spread of neural inactivation following injection of lidocaine into the monkey cerebral cortex. PMID- 9210572 TI - A simple program for simulating the responses of neurons with arbitrarily structured and active dendritic trees. AB - We describe a simple program to simulate neural responses. This program is based on the compartmental approach, in which all compartments of a neuron (i.e. axon, soma or dendrite) are represented by the same basic electrical structure. A parameter file is used to store the model parameters, including the nonlinear channel characteristics of each compartment. The model is then automatically configured according to the values specified in the parameter file. The computation of the conductance of each active channel over time is handled by a unique subroutine optimized according to the kinetics of each channel. The equations for arbitrarily structured trees are solved implicitly using a simple algorithm similar to that of Hines (Hines, M. (1984) Int. J. Biomed. Comput., 15:69-76). The output of the model uses PostScript format. The advantage of this program is that it is small in size, simple to use, efficient, and is platform independent. PMID- 9210573 TI - Rapid purification of glial cells using immunomagnetic separation. AB - By purifying glial cells from brain tissue containing a heterogeneous cell population, a number of interactions that define glial cell diversification and function within the central nervous system have been determined. The current methods for purifying glial cells, however, can be time consuming and costly. In the following study we have adapted the technique of immunomagnetic separation to separately enrich 0-2A progenitor cells and astrocytes from the rat central nervous system (CNS). In this procedure, tissue from the CNS was enzymatically dissociated and incubated in a primary antibody specific to a surface antigen found on the target cell type (e.g. A2B5 or RAN-2). The target cells were then immunologically coupled to magnetic beads, which were precoated with a secondary antibody specific to the primary, and then separated out from the heterogeneous cell population using a magnetic field. We found that the immunomagnetic separation procedure, which was completed within 2 h, produced a near pure population of glial cells (> 99%). This was confirmed by the absence of unbound cells in the bead-bound fraction. The identification and viability of bead-bound cells were established by culturing these cells and subsequently examining their morphology and antigenic expression. This study shows that glial cell types can be separated out of brain tissue to near purity using immunomagnetic separation. This simple procedure is reliable, inexpensive, and achieves levels of purity and viability comparable with currently available techniques of immunopanning and fluorescence-activated cell sorting, within a fraction of the time. PMID- 9210574 TI - Use of zonal centrifugation method for the preparation of mu-opioid receptor enriched membranes from bovine corpus striatum. AB - Earlier attempts to purify the opioid receptors met with limited success because of the use of brain crude membranes. Subcellular fractionation procedures adopted now to get enriched membranes resulted in 2-3 fold enrichment. However, a procedure has been developed in our laboratory in which a crude membrane fraction obtained at 17,500 x g was lysed with 1 mM sodium bicarbonate and later subjected to zonal fractionation, employing a density gradient of 0.6-1.2 M sucrose. This could yield a membrane fraction highly enriched with mu-opioid receptors which is 9.3 fold higher than the crude membranes. PMID- 9210576 TI - Microdialysis monitoring of extracellular glutamate combined with the simultaneous recording of evoked field potentials in hippocampal organotypic slice cultures. AB - These experiments combined extracellular electrophysiological multirecordings from hippocampal organotypic slice cultures with application of drugs to and sampling of extracellular fluid from a restricted region of the slice using a microdialysis probe. Glutamate (Glu) concentrations were monitored in 0.5 or 2 min microdialysis samples, while evoked field potentials responses (EvFPR) in the CA1 region of the hippocampus (stimulation in the CA3 area) were simultaneously recorded using a multi-electrodes array (Physiocard). Glu was assayed by capillary electrophoresis with laser-induced fluorescence detection combined with a continuous flow derivatization of dialysates. The performance of this combined approach was demonstrated by monitoring extracellular Glu concentrations and EvFPR after K+ induced depolarisation, Glu uptake blockade by trans-pyrrolidine 2,4-dicarboxylic acid (PDC), and electrical stimulation. Such an approach allows a global monitoring of the neuronal functioning with a fine time resolution (up to 30 s) on a simple in vitro brain slice model, to be used as a complement to conventional in vivo microdialysis studies. PMID- 9210575 TI - Texture analysis of cerebral white matter in SIV-infected macaque monkeys. AB - Image texture analysis is used in a wide variety of applications in medical research. Neurovirulent simian immunodeficiency virus (SIV) infection in monkeys is considered a good model for HIV-1 infection in humans and causes neuropathological changes in white matter which can include diffuse myelin pallor, subtle white matter astrocytosis, perivascular macrophage infiltrates, and microglial nodules with multinucleated giant cells. The ability of image texture analysis to quantify these changes was evaluated. Sections of thionin stained brain tissue from eight male rhesus macaques ranging in age from 42-59 months were used. Four animals served as controls and four animals were infected with neurovirulent SIVmac239/17E-R71 by bone marrow inoculation. Images of cerebral white matter were captured and analyzed by calculating 13 textural features based on statistical analysis of spatial co-occurrence matrices. Statistical analysis of the results included multiple comparisons using the Newman-Keuls multiple range test. The effect of variation in background illumination used at image acquisition was also evaluated. Ten of the 13 textural features used in this study successfully discriminated between tissue from control and SIV-infected animals and were consistent with independent neuropathological assessment. Three textural features were highly sensitive to variation in background illumination and found not useful in this application. PMID- 9210577 TI - Adaptive Fourier modeling for quantification of tremor. AB - A new computational method for quantification of tremor, the weighted frequency Fourier linear combiner (WFLC), is presented. This technique rapidly determines the frequency and amplitude of tremor by adjusting its filter weights according to a gradient search method. It provides continual tracking of frequency and amplitude modulations over the course of a test. By quantifying time-varying characteristics, the WFLC assists in correctly interpreting the results of spectral analysis, particularly for recordings exhibiting multiple spectral peaks. It therefore supplements spectral analysis, providing a more accurate picture of tremor than spectral analysis alone. The method has been incorporated into a desktop tremor measurement system to provide clinically useful analysis of tremor recorded during handwriting and drawing using a digitizing tablet. Simulated data clearly demonstrate tracking of variations in frequency and amplitude. Clinical recordings then show specific examples of quantification of time-varying aspects of tremor. PMID- 9210578 TI - Light microscopic image analysis system to quantify immunoreactive terminal area apposed to nerve cells. AB - The present report describes a desktop computer-based method for the quantitative assessment of the area occupied by immunoreactive terminals in close apposition to nerve cells in relation to the perimeter of the cell soma. This method is based on Fast Fourier Transform (FFT) routines incorporated in NIH-Image public domain software. Pyramidal cells of layer V of the somatosensory cortex outlined by GABA immunolabeled terminals were chosen for our analysis. A Leitz Diaplan light microscope was employed for the visualization of the sections. A Sierra Scientific Model 4030 CCD camera was used to capture the images into a Macintosh Centris 650 computer. After preprocessing, filtering was performed on the power spectrum in the frequency domain produced by the FFT operation. An inverse FFT with filter procedure was employed to restore the images to the spatial domain. Pasting of the original image to the transformed one using a Boolean logic operation called 'AND'ing produced an image with the terminals enhanced. This procedure allowed the creation of a binary image using a well-defined threshold of 128. Thus, the terminal area appears in black against a white background. This methodology provides an objective means of measurement of area by counting the total number of pixels occupied by immunoreactive terminals in light microscopic sections in which the difficulties of labeling intensity, size, shape and numerical density of terminals are avoided. PMID- 9210579 TI - Technique of selective spinal cord cooling in rat: methodology and application. AB - In a number of interventions, it is desirable to be able to produce a rapid but readily reversible change in spinal cord temperature (SCT) without altering general body temperature and to maintain this selective spinal cord hypothermia stable for an extended interval. To accomplish this, we developed a technique of subcutaneous perfusion cooling in rat. This was accomplished by constructing a copper heat exchanger which was readily implanted into subcutaneous space overlying the upper thoracic to upper sacral spinal segments. The heat exchanger was then perfused with fluid from an external temperature bath maintained at (8 degrees C) at a perfusion rate of 100 ml/min. The temperature of the heat exchanger was controlled by regulating the pump with a feed back controller driven by a thermocouple placed percutaneously into the paraspinal musculature. A series of studies were performed to demonstrate the characteristics and utility of this cooling technique. Lowering the pump set point to 24 degrees C resulted in a fall in the intrathecal temperature (ITT) to 27 +/- 0.3 degrees C within 15 min with no significant changes observed in rectal temperature (37.5- > 37.2 degrees C). Change in intrathecal temperature showed a highly significant correlation with changes in paravertebral muscle temperature (r = 0.977). The hypothermic state could be readily maintained for extended intervals up to 5 h and an underbody heating pad was used to maintain rectal temperature between 35 36.5 degrees C. Lowering the ITT from 37 degrees C-27 degrees C evoked a temperature-dependent increase in the latency of precooling spinal somatosensory evoked potentials (SSEPs) with the highest sensitivity observed in postsynaptic components. Returning the set point temperature back to 37 degrees C produced a rapid recovery of the SSEPs latencies. Consistent with previously published data, selective spinal cord hypothermia (27 degrees C) provided complete protection against otherwise injurious interval of normothermic ischemia produced by balloon occlusion of the descending aorta. This technique provides a simple, relatively non-invasive and reliable experimental tool for studying the effect of selective, acute and/or prolonged spinal cord hypothermia. PMID- 9210580 TI - Quantitative assessment of bradykinesia in patients with Parkinson's disease. AB - In order to optimize a method for quantitative assessment of bradykinesia, we evaluated the three-dimensional sources of a movement signal of the wrist and influence of tremor on the reliability of bradykinesia measurements. A total of 33 patients with Parkinson's disease, three patients with Multiple System Atrophy and 29 healthy controls performed a test procedure to measure slowness of movement, consisting of a tap rate (TR) test and a movement time (MT) test. Simultaneously, accelerometers were mounted on the wrist and mean bi- and tri axial vectors were calculated. Thus the acquired means of acceleration were correlated with the commonly used measures of bradykinesia. i.e. tap rate and movement time. Our results show that bradykinesia is reliably measured by the evaluation of the mean acceleration of movements, and support the use of any of the three bi-axial vectors. Compared to the bi-axial vectors, the tri-axial vector provided no relevant additional information. Additionally, the presence of a moderate to severe resting tremor did not influence the assessment of bradykinesia. Because of the possibility of continuous assessment of bradykinesia this new monitor may prove to be of great value in pharmacodynamic studies and the longitudinal follow-up of patients in drug trials. PMID- 9210581 TI - Molecular evolution of the rhodopsin gene of marine lamprey, Petromyzon marinus. AB - Visual pigment genes have been isolated from a marine lamprey, Petromyzon marinus. We report here the rhodopsin gene, spanning 21.2 kb from start to stop codons, making it the longest opsin gene known in vertebrates. Southern analysis suggests that the lamprey genome contains a single rhodopsin gene. The amino acid (aa) sequence deduced from this gene has 92% sequence similarity with that of the river lamprey rhodopsin. The data reveal that aa substitutions occurred more often in the transmembrane region than in the non-transmembrane region, possibly reflecting functional adaptation of the rhodopsin during the last 500 million years of the jawless fish evolution. PMID- 9210582 TI - Isolation and characterization of the promoter region of the chicken N-cadherin gene. AB - N-cadherin (CDH2) is a member of the cadherin family of Ca2(+)-dependent cell cell adhesion molecules. To investigate mechanisms controlling CDH2 transcription, we isolated and analyzed a genomic DNA sequence containing 2.8 kb of 5' flanking region and the first two exons of chicken CDH2. Sequence analysis of the promoter region of CDH2 revealed no CCATT or TATA boxes, but showed a high overall GC content, high CpG dinucleotide content, and several consensus Sp1 and Ap2 binding sequences. When fused to the cat reporter gene in transient transfection experiments, the sequence from positions -3231 to -118 (relative to the translation start site) directed high-level expression in CDH2-expressing chicken primary retinal cells and mouse N2A cells, but was much less active in chicken embryonic fibroblast cells and mouse 3T3 cells which do not express CDH2. Similarly, this promoter fragment directed variable, but neuronal-specific, expression of reporter genes in adult transgenic mice, but failed to produce the correct pattern of expression in other tissues, implying that additional sequences further upstream and/or within introns of CDH2 may play important roles in the transcriptional control. PMID- 9210583 TI - tap, a Drosophila bHLH gene expressed in chemosensory organs. AB - We have isolated a Drosophila bHLH gene, tap, that is expressed in a small subset of neurons when they undergo differentiation. In the peripheral nervous system, tap is expressed exclusively in one of the neurons that innervate each larval chemosensory organ, possibly controlling the specific properties of that neuron. Sequence comparisons suggest that tap is most closely related to two bHLH genes identified in several vertebrate species, neurogenin and neuroD, which are involved respectively in neural determination and in neuronal differentiation. PMID- 9210584 TI - The nuclear autoantigen La/SS-B: mapping and sequencing of the gene and the three retropseudogenes. AB - One target of autoantibodies in sera of patients with systemic lupus erythematosus or primary Sjogren's syndrome is the nuclear autoantigen La/SS-B. Lambda clones and cosmids were isolated, which contained the sequences of the La gene and the three La pseudogenes. They were used for preparation of a physical map. Finally, the La gene and pseudogenes were sequenced. The pseudogenes were characterized as retropseudogenes. Their evolutionary ages were estimated to be approx. 4, 4.5 and 5 million years. Inserts of 4, 16 and 24 nucleotides, which were mostly A-residues, were found in exon 7 of the respective pseudogene. The oldest pseudogene contained the longest insert, the youngest pseudogene contained the smallest insert. The oligonucleotides seem to be the result of repeated inserts of A-residues in a hot spot region of the La genes. Two La cDNAs were isolated which contained either a deletion or an insert of an A-residue at the same position. PMID- 9210585 TI - Structures of genes encoding phospholipase A2 inhibitors from the serum of Trimeresurus flavoviridis snake. AB - Inhibitors (PLIs) against snake venom gland phospholipases A2 (PLA2s) have been found in their sera. A cDNA encoding a PLI from Trimeresurus flavoviridis (Tf, habu snake, Crotalinae) serum, cPLI-A, was isolated from the Tf liver cDNA library and sequenced. Northern blot analysis with cPLI-A showed that PLIs are expressed only in liver. Genes for PLIs, gPLI-A and gPLI-B, were isolated from the Tf genomic DNA library and their nucleotide (nt) sequences were determined. The genes consisted of four exons and three introns, and exon 4 encoded the carbohydrate recognition domain (CRD)-like motif. Comparison of the nt sequences between gPLI-A and gPLI-B showed that these genes are highly homologous, including introns, except that exon 3 is rich in nonsynonymous nt substitutions which are almost four times as frequent as synonymous nt substitutions. This evolutionary feature of PLI genes is different from that of venom gland PLA2 isozyme genes in which nonsynonymous nt substitutions are spread over the entire mature protein-coding region. PMID- 9210586 TI - Mapping of repetitive and non-repetitive DNA probes to chromosomes of the microsporidian Encephalitozoon cuniculi. AB - The molecular karyotype of a murine isolate of Encephalitozoon cuniculi, a microsporidian with a wide range of mammalian hosts, comprises eleven chromosomes ranging in size between 217 and 315 kb. To determine specific chromosomal markers, a partial genomic library was constructed and cloned DNA fragments were hybridized to chromosomal bands separated by pulsed-field gel electrophoresis. Most probes were assigned to single chromosomes, indicating prevalence of low copy number nucleotide sequences within the very small genome of E. cuniculi (2.9 Mb). A few probes were shown to hybridize to all chromosomes. These repetitive DNA fragments corresponded to either rRNA genes or some non-coding regions whose sequences were characterized by short micro- and minisatellites. The chromosomal locations of beta-tubulin genes and six newly identified protein-encoding genes were determined. Genes encoding dihydrofolate reductase, thymidylate synthase, serine hydroxymethyl transferase, a cdc2 kinase-like protein and helicase ERCC6 like protein were each located on a single chromosome whereas genes for both beta tubulin and aminopeptidase were on two different chromosomes. The mapping will serve as a reference for further analysis of intraspecific karyotype polymorphism in different isolates from different host species. PMID- 9210587 TI - Cloning and nucleotide sequence of Bacillus stearothermophilus pyruvate carboxylase. AB - A gene for prokaryotic pyruvate carboxylase (PC) was cloned from Bacillus stearothermophilus. It has an open reading frame of 3441 base pairs which can code for a protein of 128,353 Da. Not only the molecular size and domain organization but also the deduced amino acid sequence of B. stearothermophilus PC are similar to those of eukaryotic PCs. PMID- 9210588 TI - Isolation and characterization of a cDNA clone encoding an osmotin-like protein from Arabidopsis thaliana. AB - A phage library of cDNA from Arabidopsis thaliana has been screened with oligodeoxyribonucleotides designed from regions of high homology found in tobacco osmotin and other osmotin-like proteins. One of the selected clones, Atosm34, presents a 734 bp open reading frame encoding a polypeptide of 244 amino acids, including the putative N-terminal signal and C-terminal propeptide sequences. Comparative alignment reveals extensive homologies to osmotin and the osmotin like proteins found in Solanaceae, and also to a related polypeptide found in soybean. Genomic hybridization suggests that the cDNA obtained here corresponds to a single copy gene, and RNA blot analysis showed that the level of expression is highest in old leaves. PMID- 9210589 TI - Sequence variation in the hpd gene of nonencapsulated Haemophilus influenzae isolated from patients with chronic bronchitis. AB - The molecular diversity of protein D of nonencapsulated Haemophilus influenzae strains isolated from persistently infected patients with chronic bronchitis was studied by sequencing the hpd gene of four independently obtained isolates. The nucleotide (nt) sequences of the hpd genes of two strains were identical. The other two hpd sequences showed nt substitutions which were mostly synonymous. As a consequence the deduced amino acid (aa) sequences differed from the consensus sequence only by a few aa. No changes in the hpd genes were observed among the four variants of the four strains persisting in chronic bronchitis patients for 9, 11, 8 and 3 months, respectively, although variation in their major outer membrane proteins P2 and P5 occurred. We conclude that the hpd gene is conserved during chronic infections of nonencapsulated H. influenzae. PMID- 9210590 TI - Pyruvate decarboxylase and anaerobic survival in Aspergillus nidulans. AB - The presence of pyruvate decarboxylase activity has been demonstrated in Aspergillus nidulans, and a gene encoding a pyruvate decarboxylase has been isolated from this organism and physically characterized. The isolation of the pdcA gene in A. nidulans confirms the existence of the alcoholic fermentation pathway in this fungus, despite it being an obligate aerobic organism. Southern analysis showed that it is most probably a single copy gene. Several potential binding sites for a GATAR-binding protein were identified in the sequence just prior to the start point of transcription, and mutant alleles of the GATAR binding protein-encoding gene, areA, affected pdcA mRNA levels in a manner that suggested that it influences pdcA expression in nitrogen repressing conditions. Other previously reported cases of AREA action are in nitrogen-limiting conditions. Interestingly, the production of ethanol was affected in a similar way by the same areA alleles, suggesting that changes in pdcA mRNA level are reflected in the changes in the level of ethanol production. The experiments presented here confirm that PDC levels are a major determinant of ethanol production under these conditions. PMID- 9210591 TI - Human BAC library: construction and rapid screening. AB - We have constructed a human genomic bacterial artificial chromosome (BAC) library using high molecular weight DNA from a pre-pro-B cell line, FLEB14-14, with a normal male diploid karyotype. This BAC library consists of 96,000 clones with an average DNA insert size of 110 kb, covering the human genome approximately 3 times. The library can be screened by three different methods. (1) Probe hybridization to 31 high-density replica (HDR) filters: each filter contains 3072 BAC clones which were gridded in a 6 x 6 pattern. (2) Probe hybridization to two Southern blot filters to which 31 HindIII digests of the pooled 3072 BAC clones were loaded. This identifies a particular HDR filter for which further probe hybridization is performed to identify a particular clone(s). (3) Two-step polymerase chain reaction (PCR). First, PCR is applied to DNA samples prepared from ten superpools of 9600 BAC clones each to identify a particular superpool and the second PCR is applied to 40 unique DNA samples prepared from the four dimensionally assigned BAC clones of the particular superpool. We present typical examples of the library screening using these three methods. The two-step PCR screening is particularly powerful since it allows us to isolate a desired BAC clone(s) within a day or so. The theoretical consideration of the advantage of this method is presented. Furthermore, we have adapted Vectorette method to our BAC library for the isolation of terminal sequences of the BAC DNA insert to facilitate contig formation by BAC walking. PMID- 9210592 TI - Molecular characterization of coding and untranslated regions of rat cortex lithium-sensitive myo-inositol monophosphatase cDNA. AB - Lithium sensitive myo-inositol monophosphatase (IMPase) is a pivotal enzyme which controls the levels of brain inositol within the inositol-based signaling system. Its capacity to release free myo-inositol from inositol monophosphates generated from receptor-linked and de novo pathways is crucial to the maintenance of appropriate amounts of intracellular myo-inositol, which is essential for both inositol-based cell signaling and cell volume control. We present here the full length cDNA encompassing the coding and untranslated regions (5'- and 3'-UTRs) of rat brain IMPase. This cDNA was derived from rat cortex mRNA by the RT-PCR technique. Analysis of this cDNA revealed several interesting features which include a short 5'-untranslated region (5'-UTR) of 68 nucleotides followed by coding region of approximately 0.8 kb and a long 3'-untranslated region (3'-UTR) of 1.2 kb. Both 5'-rapid amplification of cDNA ends (5'-RACE) and 3'-RACE techniques were carried out to isolate both UTRs and double stranded sequencing was carried out to its entirety (approximately 2.1 kb) by 'gene walking' using several oligonucleotide primers. All nucleotides were sequenced unambiguously using the sense and antisense strands of DNA. PCR analysis for the coding region and the deduced amino acid sequence demonstrated a DNA fragment of 831 bp and 277 amino acids, respectively, which are strikingly similar to human hippocampal IMPase. The 5'-UTR demonstrated distinct CpG doublets, characteristic of 'housekeeping' genes. The sequence around the initiator methionine, AAGATGG, conforms well to the Kozak consensus sequence for mammalian protein biosynthesis and the 3'-UTR demonstrated three canonical (AATAAT, AATTAA, AATACA) and one unusual polyadenylation signals (ATTAAA) followed by a 31 base poly(A) tail. The presence of a CCTGTG in the 3'-UTR (putative carbohydrate response element) links IMPase mRNA to brain carbohydrate metabolic pathways. Computer analyses demonstrated several unique features of this mRNA, including the potential formation of hairpin loops which might be important in its intracellular regulation and turn-over. In summary, this lithium-sensitive brain IMPase mRNA has the following characteristics: a 5'-CpG-rich short untranslated segment, a highly conserved coding region, and a long 3'-untranslated region with several polyadenylation signals. PMID- 9210593 TI - Isolation and characterization of human cDNAs encoding a cGMP-stimulated 3',5' cyclic nucleotide phosphodiesterase. AB - Human cyclic GMP-stimulated 3',5'-cyclic nucleotide phosphodiesterase (PDE2A3) cDNAs were cloned from hippocampus and fetal brain cDNA libraries. A 4.2-kb composite DNA sequence constructed from overlapping cDNA clones encodes a 941 amino acid protein with a predicted molecular mass of 105,715 Da. Extracts prepared from yeast expressing the human PDE2A3 hydrolyzed both cyclic AMP (cAMP) and cyclic GMP (cGMP). This activity was inhibited by EHNA, a selective PDE2 inhibitor, and was stimulated three-fold by cGMP. Human PDE2A is expressed in brain and to a lesser extent in heart, placenta, lung, skeletal muscle, kidney and pancreas. The human PDE2A3 differs from the bovine PDE2A1 and rat PDE2A2 proteins at the amino terminus but its amino-terminal sequence is identical to the bovine PDE2A3 sequence. The different amino termini probably arise from alternative exon splicing of the PDE2A mRNA. PMID- 9210595 TI - Isolation of transforming growth factor-beta2 cDNA from a fish, Cyprinus carpio by RT-PCR. AB - Transforming Growth Factors-beta (TGF-betas) have been described in many vertebrate species of amphibians, aves and mammals. In this report we demonstrate the presence of TGF-beta2 in pisces. TGF-beta2 has been cloned from a fish, Cyprinus carpio, by RT-PCR using degenerate oligonucleotide primers. Sequence analysis of the amplified product and alignment of the deduced amino acid sequence with the human TGF-beta2 amino acid sequence revealed 81% and 93% identity in the precursor and the mature regions, respectively. The northern blot analysis of fish heart RNA shows a major messenger RNA species of about 8.0 kb and two messages of very low abundance of about 5.0 kb and 4.0 kb. The identification of TGF-beta2 isoform in Pisces and it's high degree of homology with the mammalian isoform suggests that among all TGF-beta isoforms, TGF-beta2 is the most conserved during evolution. PMID- 9210594 TI - Molecular cloning of cDNA for lysenin, a novel protein in the earthworm Eisenia foetida that causes contraction of rat vascular smooth muscle. AB - Lysenin, which causes contraction of rat vascular smooth muscle, is a protein that was isolated from the earthworm Eisenia foetida. A cDNA encoding lysenin was isolated by use of a partial cDNA probe that had been generated by the PCR with a primer designed by reference to an internal peptide sequence of lysenin. This clone had an ORF encoding 297 amino acid residues. The amino acid sequence deduced from the cDNA revealed the absence of any significant homology to those of previously characterized vasoactive substances. The recombinant lysenin was produced in Escherichia coli. This protein and native lysenin isolated from the earthworm had similar contractive activities when tested on rat aorta. Northern blot analysis of the RNA from various tissues of the earthworm indicated that lysenin is produced by the coelomocytes. PMID- 9210596 TI - Cloning and characterization of the mouse tob gene. AB - The human Tob protein was identified as a molecule that interacted with the ErbB 2 protein, a receptor-type protein tyrosine kinase (RPTK). The interaction suggests that Tob is involved in RPTK-mediated signaling. In the present paper, we report molecular cloning and characterization of the mouse tob gene. The mouse tob gene contains an open reading frame of 1089 bp with 87% identity to its human counterpart in the nucleotide sequence. The coding region of mouse tob as well as human tob is not interrupted by introns. The mouse tob transcript is 2.3 kb long, the size being similar to that of the human tob transcript, and is detected ubiquitously in various tissues. Like human Tob, mouse Tob is characterized by the presence of a sequence rich in proline and glutamine which is often present in the sequence of transcription factors. In addition, the ATTTA motif characteristic of the immediate early gene is present in the 3'-untranslated region of the mouse tob gene. This, together with the conserved sequence and expression pattern of tob between mouse and human, suggests that Tob plays an important role in the response to extracellular signals. PMID- 9210597 TI - A site-specific DNA endonuclease specified by one of two ORFs encoded by a group I intron in Dictyostelium discoideum mitochondrial DNA. AB - The second intron (DdOX1/2.2) of Dictyostelium discoideum cytochrome oxidase subunit 1/2 fused gene has two free-standing ORF genes (Dd ai2a and Dd ai2b) in a loop, which have similar amino acid sequences and are homologous to aI4 DNA endonuclease (I-SceII) of Saccharomyces cerevisiae. To elucidate the functions of these ORFs, we cloned the ORFs into an expression vector and introduced the composite vectors into E. coli. The expression of Dd ai2a in E. coli caused growth inhibition and degradation of the E. coli genomic DNA. To determine whether Dd ai2a protein is a homing type DNA endonuclease, the ability to cleave the homing site of its intron in vivo was examined. Dd ai2a cleaved only one strand of intronless DNA sequence at the site which coincides with the I-SceII cleavage recognition site. We suppose that Dd ai2a functions actually as a homing type DNA endonuclease in D. discoideum mitochondria by virtue of other factors. To obtain further information about the relationship between the existence of introns and the mating system, we carried out in vitro self-splicing assay and polymerase chain reaction analysis using 13 strains of the cellular slime mold. PMID- 9210598 TI - Highly conserved 5'-flanking regions of two self-incompatibility genes, SLG9 and SRK9. AB - The nucleotide (nt) sequences of the 5'-flanking regions of two Brassica self incompatibility genes, SLG9 and SRK9, were determined. Their sequences were highly conserved: a region spanning 1.9 kb in the 5'-flanking region was completely identical except for a 1319-bp segment in SLG9. These observations strongly suggest that SLG9 and SRK9 together with their promoter regions were involved in a gene duplication or conversion event which occurred before the 1319 bp SLG9-specific sequence was inserted in SLG9 or deleted in SRK9. PMID- 9210599 TI - Representation of function: the next step. PMID- 9210600 TI - Clinical implications of GBV-C/HGV infection in patients with "HCV-related" chronic hepatitis. AB - BACKGROUND/AIMS: To evaluate the clinical, biochemical and histological implications of a concomitant HGV infection in "HCV-related" chronic liver disease. METHODS: Eighty-three HCV-RNA positive patients with chronic liver disease were tested for GBV-C/HGV coinfection by heminested PCR. RESULTS: Twenty two (26.5%) patients were found to be positive for GBV-C/HGV RNA. GBV-C/HGV+ patients differed significantly from GBV-C/HGV- ones for younger age, higher frequency of history of drug addiction, which in turn might favor coinfection with interferon-sensitive HCV genotypes (3a), and increased probability of long term response to interferon. GBV-C/HGV infection appears to have no responsibility for specific aspects of HCV infection such as biochemical or histological cholestatic features, lymphoid follicles, symptomatic cryoglobulinemia or presence of serum autoantibodies, including LKM1. It does not worsen the HCV-related disease (ALT levels and histological activity) and does not significantly interfere with HCV infection, as explored by the number of hepatocytes positive for HCV antigens. The amount of steatosis (mean score) was shown to be higher in GBV-C/HGV+ patients. A virological follow up was performed in 17 interferon-treated GBV-C/HGV+ patients On the whole, GBV-C/HGV seems to be as sensitive to IFN treatment as HCV, but recurrence after withdrawal is more frequent. In spite of this, ALT levels often remain normal after treatment withdrawal. CONCLUSIONS: The present data suggest that GBV-C/HGV infection, apart from more marked liver steatosis, does not modify the overall picture of chronic hepatitis due to HCV infection. PMID- 9210601 TI - HCV genotypes in patients with liver disease of different stages and severity. AB - AIMS/MATERIAL: Hepatitis C virus (HCV) genotyping was performed in 213 anti-HCV positive patients with chronic liver disease ranging from minimal histological changes to hepatocellular carcinoma. One hundred and twenty-two patients had non cirrhotic chronic active or persistent hepatitis (including 29 who were asymptomatic with persistently normal ALT levels) (chronic liver disease group). The other 91 had hepatocellular carcinoma and, in all but three cases, cirrhosis (hepatocellular carcinoma group). RESULTS: The overall prevalence of HCV variants was: 54.9% type 1b, 37.8% type 2, 2.5% type 1a, 2.0% type 3a, 2.0% type 4a. The genotype distribution showed no relation to the stage (chronic liver disease vs. hepatocellular carcinoma) or severity (chronic active vs. chronic persistent hepatitis) of the liver disease, or to the duration of the disease (<10 years vs. >10 years). Within the hepatocellular carcinoma group, the duration of type-1b disease was similar to that of type-2 infections. Ages at the time of infection and genotype were both independently associated with progression to cirrhosis and hepatocellular carcinoma, but multivariate analysis revealed that the effect of age was much stronger than that of genotype 1b. CONCLUSIONS: The predominance of HCV type 1b in this study reflects the higher frequency of this variant in our area. Our findings indicate that infections caused by each HCV genotype are capable of progressing to hepatocellular carcinoma. PMID- 9210603 TI - The effect of interferon on the liver in chronic hepatitis C: a quantitative evaluation of histology by meta-analysis. AB - BACKGROUND/AIMS: Several randomized clinical trials of interferon in chronic hepatitis C have examined the histological changes in paired biopsy specimens. We have attempted a quantitative evaluation by meta-analysis. METHODS: Randomized Clinical Trials found by MEDLINE search were included if: a) they compared different IFN regimens with non-active treatment or with each other, b) they obtained biopsies before starting and at the time of stopping IFN in a sizable proportion of the treated and control patients, and c) they assessed the biopsy specimens semi-quantitatively according to Scheuer's numerical scoring system or Knodell's Histological Activity Index, with quantitation of fibrosis and of lobular, portal and periportal necroinflammation. RESULTS: Seventeen trials were identified, in which 1223 adult patients had been studied. All trials homogeneously pointed towards a favorable interferon effect. The pooled data show a statistically significant histological improvement in treated patients as compared with controls for each of the four Histological Activity Index components and for the total Histological Activity Index score (overall improvement was -0.82 in favor of interferon, p<0.0001, 95% Confidence Interval 1.25 to -0.40). In the ten trials reporting histological changes separately in biochemical responders (primary and sustained responders) and non-responders, histological improvement was confined to the subset of biochemical responders. No change or very little change occurred in non-responders. CONCLUSIONS: Interferon treatment in chronic hepatitis C significantly improves liver histology. The effect of interferon is closely related to biochemical response. Studies assessing histological outcome 1 year or more after interferon treatment in long term responders and comparatively in non-responders or relapsers would be important to confirm the regression of the necroinflammatory process in the former, as suggested by the normal serum alanine aminotransferase levels. PMID- 9210602 TI - Evaluation of hepatitis C virus envelope proteins expressed in E. coli and insect cells for use as tools for antibody screening. AB - BACKGROUND/METHODS: The two envelope proteins of hepatitis C virus, E1 and E2, were expressed in E. coli and, as secretory proteins, in Sf9 insect cells using recombinant baculoviruses. Co-infection of insect cells with E1 and E2 recombinant baculoviruses was performed, which has been shown to result in formation of E1-E2 dimers. All envelope proteins were purified by Ni2+-NTA chromatography and used for screening of serum samples in a HCV EIA assay. Serum samples of normal blood donors, chronically HCV-infected patients, a mixed titer panel and several seroconversion panels were screened and compared to test results with Cobas Core Anti-HCV EIA. RESULTS: Screening of the sera of chronically HCV-infected patients (100% positive in Cobas Core Anti-HCV EIA) revealed 10-40% anti-E1 positive sera using different Sf9-expressed, glycosylated proteins and 93% using E. coli-expressed, non-glycosylated E1 protein. When the same sera were tested with different E2 proteins expressed in Sf9 cells and in E. coli, about 70-73% showed anti-E2 reactivity. When the proteins from Sf9 cells co infected with E1- and E2-recombinant baculoviruses were tested, 70-80% of the same sera showed anti-envelope reactivity. CONCLUSIONS: Testing of these patient antisera, and those from the well-characterized mixed titer panel BBI-PHV203, showed that recombinant E1 expressed in E. coli and co-expressed E1 and E2 proteins from Sf9 cells could be used as additional tools for anti-HCV antibody screening. PMID- 9210604 TI - Hepatitis C virus infection of salivary gland epithelial cells. Lack of evidence. AB - BACKGROUND: Hepatitis C virus genome (HCV-RNA) has been detected in whole salivary gland tissue of chronically infected patients. However, contamination of the tissue by plasma or blood cells was not excluded by the previous reports. AIMS: To assess whether HCV infects the salivary gland epithelial cells in patients with chronic HCV liver disease. METHODS: Twenty unselected patients with chronic active hepatitis (11 cases) or active cirrhosis (nine cases) were examined. Serum and saliva samples were obtained from all patients, 12 of whom (seven, chronic active hepatitis; five, active cirrhosis) underwent salivary gland biopsy. PCR for HCV-RNA was performed on RNA extracted from serum, saliva and salivary gland epithelial cells collected by isokinetic gradient separation after trypsin digestion of whole salivary gland tissue. Saliva samples were also examined for the presence of secretory IgA anti-HCV by gel chromatography and ELISA testing. RESULTS: HCV-RNA was detected in all sera with titers ranging from 5.42 x 10(5) genome equivalents/ml to 123.2 x 10(5) genome equivalents/ml. Thirteen patients were infected with genotype 1b, four patients had genotype 1a, two patients had genotype 2a and one patient was unclassifiable. Low titer HCV RNA (<2 x 10(5) genome equivalents/ml) was detected in 3/20 saliva samples (15%) from highly viremic patients infected with 1b genotype. RNA extracted from salivary gland epithelial cells consistently tested negative for HCV-RNA. In addition, all saliva specimens tested negative for secretory-IgA (S-IgA) anti HCV, even after a 10-fold concentration of the samples. CONCLUSIONS: There was no evidence that HCV infects the salivary gland epithelial cells in our viremic patients with HCV chronic liver disease. Low level HCV-RNA in saliva is most probably due to virus spillover from blood. PMID- 9210605 TI - Present status of autoimmune hepatitis in Japan--correlating the characteristics with international criteria in an area with a high rate of HCV infection. Japanese National Study Group of Autoimmune Hepatitis. AB - BACKGROUND/AIMS: A nationwide survey of autoimmune hepatitis (AIH) was carried out in Japan. METHODS: Four hundred and ninety-six patients were enrolled by questionnaires sent to 101 hospitals with hepatology specialists. RESULTS: The clinical features of Japanese AIH were as follows: most patients were middle-aged women; serum autoantibodies, especially antinuclear antibody, were frequently positive, serum IgG level was high, and HLA-DR4 was the major HLA allotype. Liver kidney microsomal type 1 antibody was positive in nine of 79 patients tested. Eight of these antibody positive patients were also positive for antinuclear antibody and five for anti-smooth muscle antibody. Ninety-two percent of the patients showed piecemeal necrosis and 60% bridging necrosis; plasma cell infiltration in the portal areas was observed in 50% of the patients. Only 12.3% were diagnosed as having liver cirrhosis. A favorable effect of corticosteroid, normalization of serum transaminases, was observed in 89% of 317 patients, who were treated with an initial dose of over 30 mg/day. Sixty-two patients were positive for hepatitis C virus (HCV) markers. In these patients, however, only one patient was liver-kidney microsomal type 1 antibody positive. Corticosteroid was effective in 30 (81%) of 37 HCV-marker-positive patients treated with this agent. Thus the efficacy of corticosteroid did not differ from that in AIH patients without HCV infection (90%). Similarly, interferon treatment was used in 20 patients, all of whom were positive for HCV-RNA, and resulted in 50% efficacy as determined by normalization of the serum transaminase level 6 months after treatment. The International Diagnostic Scoring System for the diagnosis of AIH worked well in these patients, except for HCV-infected individuals, that is, approximately 10% of the total of AIH patients. PMID- 9210606 TI - Tissue inhibitor of metalloproteinase-1 in the liver of patients with chronic liver disease. AB - BACKGROUND/AIMS: Tissue inhibitor of metalloproteinase (TIMP)-1 is an important regulator of matrix metalloproteinase activity. To clarify the changes in TIMP-1 in diseased livers, we measured TIMP-1 concentrations in liver tissue samples from patients with chronic liver disease. The relationship between serum and liver levels of TIMP-1 was also examined in some patients. METHODS: The subjects were 68 patients who underwent liver biopsy. The liver TIMP-1 concentration was measured using an enzyme immunoassay after the extraction of TIMP-1 with 2 M guanidine. RESULTS: As compared with the controls (n=10), the liver TIMP-1 level was increased 2.2-fold in the 24 chronic active hepatitis 2A patients, 2.9-fold in the 10 chronic active hepatitis 2B patients and 4.1-fold in the six liver cirrhosis patients, but no significant increase was observed among the 18 chronic persistent hepatitis patients. The liver TIMP-1 levels were closely correlated with the histological degrees of periportal necrosis, portal inflammation, and liver fibrosis. When the localization of TIMP-1 was examined immunohistochemically, TIMP-1 was stained mainly in hepatocytes, and the intensity was stronger in the livers of chronic active hepatitis and liver cirrhosis patients than in those of the chronic persistent hepatitis patients. The serum TIMP-1 and liver TIMP-1 levels were significantly correlated, indicating that serum TIMP-1 could reflect the change of liver TIMP-1 in patients with chronic liver disease. CONCLUSION: Liver TIMP-1 concentration increases with progression of the liver disease, when the degradation of extracellular matrix proteins is decreased, resulting in the development of liver fibrosis. PMID- 9210607 TI - Transforming growth factor-beta-induced collagen synthesis by human liver myofibroblasts is inhibited by alpha2-macroglobulin. AB - BACKGROUND: Transforming growth factor-beta (TGFbeta) plays a central role in the stimulation of matrix production during liver fibrosis. The action of TGFbeta in different systems has been shown to be influenced by alpha2-macroglobulin (alpha2M), a serum protein with strong protease-scavenging and cytokine-binding properties. AIMS: In the present study, alpha2M derived from normal human plasma has been tested for its ability to modulate the TGFbeta-induced collagen production by human liver fat-storing cells (FSC), which had transformed into alpha-smooth muscle actin-expressing myofibroblasts in culture. METHODS: Alpha2M has been tested after activation with methylamine (alpha2M-Me), an in vitro equivalent of protease activated alpha2M. The binding of 125I-TGFbeta1 to activated forms of alpha2M was demonstrated by rate electrophoresis. Collagen synthesis was examined in human liver myofibroblast cultures obtained from three different human livers by incorporation of 3H-proline into TCA-precipitable, specific collagenase degradable proteins. Uptake of alpha2M was studied by means of immunofluorescence. RESULTS: TGFbeta (1 ng/ml) significantly stimulated collagen synthesis of controls in the absence of TGFbeta. Alpha2M-Me reduced this TGFbeta-induced collagen synthesis dose-dependently, reaching significant inhibition from 10 microg/ml alpha2M-Me onward. Upon addition of 100 microg/ml alpha2M-Me the effect of TGFbeta was reduced by 60% to 128+/-31% (mean+/-SD) of control values in the absence of TGFbeta. Human liver myofibroblasts endocytosed alpha2M-Me added to the cultures as detected by immunofluorescence. Accordingly, reduction of TGFbeta-activity by alpha2M-Me may be explained by receptor-mediated clearance of alpha2M-TGFbeta complexes by the cells. CONCLUSIONS: TGFbeta-induced collagen formation by human liver myofibroblasts obtained from three different livers is reduced in vitro by activated alpha2M. From these results, we hypothesize that alpha2M may have an antifibrogenic effect in vivo by interference with TGFbeta-induced matrix synthesis during liver fibrosis. PMID- 9210608 TI - Natriuretic response to the combination of atrial natriuretic peptide and terlipressin in patients with cirrhosis and refractory ascites. AB - BACKGROUND/AIMS: Refractory ascites, which occurs in certain patients with cirrhosis, is associated with a blunted natriuretic response to exogenous atrial natriuretic peptide (ANP). Since this blunting seems to be related to ANP-induced arterial hypotension, a vasoconstrictor, such as terlipressin (a vasopressin analogue), may restore natriuresis to exogenous ANP. Moreover, since cirrhosis elicited vasodilation is thought to play a role in sodium retention, a vasoconstriction caused by terlipressin alone may lead to an increase in sodium excretion. This study aimed to evaluate the natriuretic response to either a combination of ANP with terlipressin or terlipressin alone in patients with cirrhosis and refractory ascites. METHODS: Sixteen consecutive patients with cirrhosis and refractory ascites were randomly assigned to receive either a combination of terlipressin (1-2 mg, i.v. bolus) with ANP (35 ng/kg, i.v. bolus followed by 15 ng x kg(-1) x min(-1) for 60 min) (n=8) or terlipressin alone (1-2 mg, i.v. bolus) (n=8). Sodium excretion and urine output, systemic, splanchnic and renal hemodynamics and renal oxygen consumption were measured before and during treatments. RESULTS: Combined therapy did not change arterial pressure but significantly increased urinary sodium excretion and urine output. These effects were associated with a significant increase in glomerular filtration rate and a decrease in renal oxygen consumption. Terlipressin alone significantly increased arterial pressure but did not change urinary sodium excretion or urine output. Moreover, terlipressin did not change either glomerular filtration rate or renal oxygen consumption. CONCLUSIONS: The combination of exogenous ANP with terlipressin, but not terlipressin alone, increases sodium excretion in patients with cirrhosis and refractory ascites. PMID- 9210609 TI - Portal-hypertensive gastropathy develops less in patients with cirrhosis and fundal varices. AB - BACKGROUND/AIMS: The aim of this prospective study was to examine the association of portal-hypertensive gastropathy and fundal varices in patients with cirrhosis. METHODS: We carried out an endoscopic observation in 476 patients with cirrhosis (study 1), including 62 patients undergoing endoscopic obliteration of esophageal varices (study 2). In study 1, patients were classified into five subgroups: no esophagofundal varices (n=119), small esophagofundal varices (n=127), dominant esophageal varices (n=177), dominant fundal varices (n=27), and large esophagofundal varices (n=26). The severity of liver dysfunction was assessed by Pugh-Child classification: class A (n=222), class B (n=200), and class C (n=54). In study 2, two groups, poorly developed fundal varices (n=50) and well developed fundal (n=12), were distinguished and the follow-up endoscopic examinations were performed on the basis of 3-month intervals for 2 years. In each study, the severity of portal-hypertensive gastropathy was scored: 0 (absent), 1 (mild), 2 (severe), and 3 (bleeding). RESULTS: Study 1: One-way ANOVA showed that both variceal pattern and Pugh-Child class significantly influenced portal hypertensive gastropathy score. However, two-way ANOVA indicated that variceal pattern was the only significant variable. Portal-hypertensive gastropathy score was significantly higher in patients with dominant esophageal varices than in either patients with no esophagofundal varices or patients with small esophagofundal varices. In contrast, portal-hypertensive gastropathy score in patients with dominant fundal varices was similar to that in patients with no esophagofundal varices and was significantly lower compared with that in patients with dominant esophageal varices. Furthermore, portal-hypertensive gastropathy score was significantly lower in patients with large esophagofundal varices than in patients with dominant esophageal varices. Study 2: After the obliteration of esophageal varices, portal-hypertensive gastropathy score in patients with poorly developed fundal varices became significantly higher at 3-, 6-, 9-months while it was not modified in patients with well developed fundal varices during the follow up period. Furthermore, the integrated incremental change in portal-hypertensive gastropathy score during the first 1-year follow-up period was significantly lower in patients with well developed fundal varices than in patients with poorly developed fundal varices. CONCLUSIONS: These results indicate that both spontaneous and obliteration-induced portal-hypertensive gastropathy lesions develop less in patients with cirrhosis and fundal varices. PMID- 9210610 TI - Autonomic dysfunction in patients with non-alcoholic chronic liver disease. AB - AIMS: To assess the presence of autonomic neuropathy in patients with non alcoholic chronic liver disease and its relationships with the severity of liver damage. METHODS: Thirty non-alcoholic patients with chronic liver disease and 26 healthy control subjects were studied. The silicone imprint technique was used to quantify the number of functioning sweat glands in order to assess peripheral sympathetic dysfunction. Heart rate variations in response to deep breathing at 6 per minute (deltaR6), to a Valsalva maneuver, and with orthostatism (RRmax/RRmin) were determined to assess parasympathetic vagal function. RESULTS: Mean values for autonomic tests were significantly lower in the group of patients with non alcoholic chronic liver disease than in the control subjects. The number of activated sweat glands in the foot was abnormal in 19 (63%) patients. Among vagal tests, Valsalva ratio was abnormal in 14 (46%) and deltaR6 in 11 (36%) patients with liver disease. Vagal neuropathy (two or more abnormal heart rate tests) was definite in nine patients (30%). CONCLUSIONS: A high prevalence of abnormalities in both sympathetic and parasympathetic function tests, with a poor relationship with liver function parameters, has been found in patients with non-alcoholic chronic liver disease. PMID- 9210611 TI - Seroprevalence and epidemiology of Helicobacter pylori infection in patients with cirrhosis. AB - BACKGROUND: Helicobacter pylori infection is the major pathogenic factor for peptic ulcer disease. Its epidemiology is not fully known; few data are available in patients with chronic liver disease. AIMS: To investigate the seroprevalence and factors associated with Helicobacter pylori infection in a series of liver cirrhosis patients. METHODS: Two hundred and twenty consecutive patients were prospectively included in a study aimed to evaluate the effect of dietary intervention on cirrhosis complications and survival. At inclusion, an epidemiological and clinical questionnaire was completed. Sera were obtained and stored at -70 degrees C until analyzed. They were tested for Helicobacter pylori antibodies using a commercial ELISA kit. RESULTS: Eleven out of 220 patients had borderline anti-Helicobacter pylori IgG titers. Of the remaining 209 patients, 105 (50.2%) showed positive titers of Helicobacter pylori IgG. Univariate analysis showed that Helicobacter pylori infection was more frequent in older patients, those born outside Catalonia, and in patients with a low educational level. Past ethanol consumption and current smoking correlated negatively with Helicobacter pylori infection. Multivariate analysis selected age (OR 3.1. 95% CI 1.46-6.45), educational level (OR 2.2. 95% CI 1.18-4.2) and alcohol consumption (OR 0.7. 95% CI 0.45-0.99) as the variables independently related to Helicobacter pylori infection. CONCLUSIONS: Helicobacter pylori infection in cirrhosis has the same epidemiological pattern as in the general population. Suggestions that the etiology or the severity of the liver disease could be related to Helicobacter pylori infection were not confirmed by our study. PMID- 9210612 TI - Diversity of antinuclear antibody responses in hepatocellular carcinoma. AB - BACKGROUND/AIMS: The development of antinuclear antibodies (ANA) in malignancies has been described but its mechanism is still not understood. The aim of this study was to examine ANA specificities in hepatocellular carcinoma to further understand autoimmunity in cancer. METHODS: Two hundred and four hepatocellular carcinoma patients were compared with 68 chronic hepatitis C, with 126 chronic hepatitis B and with 30 alcoholic liver cirrhosis patients, as well as with 87 healthy donors. Indirect immunofluorescence, immunoblotting, and immunoprecipitation were used to study ANA reactivities. RESULTS: Hepatocellular carcinoma had a significantly higher frequency of ANA using HEp-2 cells as substrate (31%) than chronic hepatitis C (10%), chronic hepatitis B (9.5%), alcoholic liver cirrhosis (10%) or healthy donors (4.5%). A great diversity of ANA specificities was found in hepatocellular carcinoma. Three hepatoma sera had antibodies that co-localized with non-snRNP splicing factor SC35, suggesting that the antigenic targets might be involved in mRNA splicing. We identified antibodies to two known nuclear autoantigens: fibrillarin and p330d/CENP-F. These autoantigens are involved in the 5' processing of precursor ribosomal RNA transcripts and in mitotic functions, respectively. CONCLUSIONS: Diversity was found in the autoantibody specificity, in contrast to the specific subsets of autoantibodies seen in several systemic rheumatic autoimmune diseases. Our data suggest that immune response in hepatocellular carcinoma targets important proteins involved in cellular biosynthetic or proliferative functions. PMID- 9210613 TI - Increased ubiquitin immunoreactivity in hepatocellular carcinomas and precancerous lesions of the liver. AB - BACKGROUND/AIMS: Ubiquitin covalently attaches to abnormal and short-lived proteins, thus marking them for ATP-dependent proteolysis in eukaryotic cells. Increased ubiquitin immunoreactivity was recently observed immunohistochemically in human malignant tumors. To clarify the change in protein metabolism during hepatocarcinogenesis, we studied ubiquitin immunoreactivity in hepatocellular carcinomas (HCCs) and precancerous lesions using immunohistochemistry and immunoblot analysis. METHODS: A total of 72 HCCs (37 advanced, 19 early, 16 early advanced (advanced HCC component in early HCC nodule) type HCCs) and 18 precancerous lesions (8 atypical adenomatous hyperplasias (AAHs), 10 adenomatous hyperplasias (AHs)) were studied immunohistochemically. Immunoblot analysis was also performed in advanced HCC and early HCC cases. RESULTS: Non-tumorous hepatocytes were either immunonegative or weakly stained in their nuclei. Advanced HCCs showed strong immunoreactivity in most cases, while early HCCs showed relatively weaker immunoreactivity. In 14 of 16 early-advanced type tumors, the inner portion of the nodules, which corresponds to advanced HCC, showed stronger immunoreactivity than the outer low-grade portion. In 8 of 8 AAHs and 7 of 10 AHs, positive but weak staining was found. Immunoblot analysis showed an increase in 42 kDa ubiquitinated protein(s) in 8 of 16 advanced HCC cases (50%) and in 1 of 6 early HCC cases (16.7%), as well as an increase in several other bands in tumor tissues. CONCLUSIONS: The intensity of ubiquitin staining appeared to increase in a stepwise manner from AH to advanced HCC, and the results suggest a possible correlation between changes in the ubiquitinated proteins and multistep hepatocarcinogenesis. PMID- 9210614 TI - Liver resection or transplantation for hepatocellular carcinoma? Retrospective analysis of 215 patients with cirrhosis. AB - BACKGROUND/AIMS: Currently, surgical treatment of hepatocellular carcinoma in patients with cirrhosis is not clearly defined. The objective of this study was, in patients with cirrhosis with hepatocellular carcinoma, to compare liver resection to transplantation assessed by patient survival and to determine whether the tumor recurrence might be influenced by prognostic factors. METHODS: We have gathered all the available data from six French Medical Universities, for 215 patients with cirrhosis with hepatocellular carcinoma surgically treated either by liver resection (102) or by transplantation (113). RESULTS: The overall 5-year survival rate was similar in the transplantation group and in the resection group (32% vs. 31%, p=0.7). However, the 5-year survival rate without recurrence was higher in the transplantation group than in the resection group (60% vs. 14%, p<0.001). Three independent prognostic factors influenced significantly the survival without recurrence: the surgical treatment by transplantation (p<0.001), the number of tumors (p<0.01) and the tumor size (p<0.001). With these factors we defined a prognostic index (Ip) which allowed assessment of the probability of survival without recurrence: Ip= (Xie. x 1.41)+(Nbr T. x 0.19)+(Size TV. x 0.16); Xie=surgical treatment (Xie=0 if transplantation, Xie=1 if resection), Nbr.T. and Size TV.=number of tumors and size of the most voluminous tumor, respectively, according to the histologic study. CONCLUSIONS: These results and this prognostic index are encouraging for liver transplantation as treatment of hepatocellular carcinoma in selected patients with cirrhosis. PMID- 9210615 TI - NHE-3 isoform of the Na+/H+ exchanger in human gallbladder. Localization of specific mRNA by in situ hybridization. AB - BACKGROUND/AIMS: Electroneutral absorption of NaCl by the gallbladder mucosa is likely to depend at least in part on a Na+/H+ exchanger. In intestine and colon, absorption due to Na+/H+ exchanger is explained by the presence of specific isoforms of the exchanger, the NHE-3 isoform and possibly the NHE-2 isoform. The aim of the present work was to determine whether the mRNAs coding for NHE-2 and NHE-3 are expressed in epithelial cells of human gallbladder. METHODS: Total RNAs from human gallbladder were subjected to reverse transcription-polymerase chain reaction using specific primers. No message was observed with NHE-2 specific primers, showing that NHE-2 isoform plays no role in gallbladder absorption. With NHE-3 specific primers, a 239 bp cDNA fragment was obtained and showed a high homology with the NHE-3 isoform, confirming the presence of NHE-3 in the gallbladder wall. This fragment was cloned in a pLitmus vector in order to produce cRNA probes by in vitro transcription. Cellular localization of the NHE-3 mRNA was studied on cryostat sections using the cRNA probes labeled with Digoxigenin-11-UTP, controls included assays with sense probe, antibodies without probe and RNaseA treated tissue. A specific staining of the NHE-3 mRNAs was found to be strictly localized to the gallbladder epithelial cells. RESULTS/CONCLUSIONS: Expression of NHE-3 in the gallbladder was found only in the absorptive epithelial cells. The NHE-3 isoform of the Na+/H+ exchanger is likely to be involved in water and electrolyte absorption from bile. PMID- 9210616 TI - Patterns of intermediate filaments, VLA integrins and HLA antigens in a new human biliary epithelial cell line sensitive to interferon-gamma. AB - BACKGROUND/AIMS: Intra-hepatic bile ducts are the primary site of damage in several immunologically mediated liver diseases. However, immunological processes underlying biliary epithelial cell recognition by T lymphocytes are poorly understood. Therefore, a convenient in vitro model that could mimic these immunologic disorders would be of great interest. METHODS: A human cell line (HuGB) was established from a metastasis of gallbladder adenocarcinoma in the liver. Intermediate filament expression was analysed by immunostaining, and gamma glutamyl transpeptidase and albumin secretion were measured. VLA integrin expression pattern, expression of HLA class I and II antigens and ICAM-1 protein were analysed by flow cytometry and their modulation by interferon-gamma was quantitated using a QIFIKIT commercial kit. RESULTS: Histological analysis showed high similarity between the initial gallbladder adenocarcinoma and the established cell line. Cytokeratins 8 and 19 and vimentin showed strong positive staining in the established cell line. Gamma-glutamyl transpeptidase was secreted by these cells while albumin expression was negative. HuGB cells also expressed VLA-alpha2, VLA-alpha3, VLA-alpha6, VLA-beta1, but not VLA-alpha1, VLA-alpha4 and NCAM, a pattern of adhesion molecule expression compatible with the biliary epithelium. Also, similar to the biliary epithelium found in normal liver, HuGB cells expressed abundant HLA class I but few HLA class II antigens. We found that the expression of HLA antigens and ICAM-1 protein were increased during interferon-gamma treatment of HuGB cell line. CONCLUSIONS: Both phenotypic and morphological characteristics of HuGB cells suggested their biliary origin. Sensitivity of HuGB cells to interferon-gamma suggests that this new cell line could represent a suitable model to investigate the up-regulation of membrane antigens occurring in immune diseases involving biliary epithelial cells. PMID- 9210617 TI - Gallbladder motility and lithogenicity of bile in patients with choledocholithiasis after endoscopic sphincterotomy. AB - BACKGROUND/AIMS: Ablation of the sphincter of Oddi has been shown to inhibit gallstone formation in the prairie dog model, probably by alleviating gallbladder bile stasis. The effect of endoscopic sphincterotomy (ES) on gallbladder emptying and lithogenicity of bile has not been studied adequately in humans. We, therefore, studied the changes in gallbladder emptying and lithogenicity of bile following ES in patients with choledocholithiasis and gallbladder in situ. METHODS: Thirteen patients with choledocholithiasis with intact gallbladder underwent ES and common bile duct clearance. Eight patients had concomitant gallstones. Gallbladder emptying was studied by real time ultrasonography after stimulation by ceruletid infusion. Fasting gallbladder bile was collected during endoscopic retrograde cholangiography by placing a 7F or 8F catheter in the common bile duct and after ceruletid stimulation of gallbladder for bile microscopy and cholesterol nucleation time determination. Gallbladder emptying, nucleation time and bile microscopy were performed before ES and again between 4 and 8 weeks after ES after cholangiographic confirmation of clearance of common bile duct stones. RESULTS: Fasting and residual gallbladder volumes decreased and ejection fraction increased significantly following ES, suggesting decreased stasis and improved emptying of gallbladder. Nucleation time was prolonged and cholesterol crystal index in bile decreased after ES, suggesting decreased lithogenicity. The decrease in gallbladder volumes and increase in ejection fraction after ES were observed in both groups of patients, with or without concomitant gallstones. CONCLUSIONS: ES decreases the stasis of gallbladder bile, improves gallbladder emptying and decreases the lithogenicity of bile in patients with gallstone disease as reflected by prolongation in nucleation time. ES may find a role as an adjunct to oral bile acid therapy and extracorporeal shock wave lithotripsy in addition to a prophylactic role of preventing gallstone formation in high risk groups. PMID- 9210618 TI - Cholesterol metabolism and non-cholesterol sterols in patients with ileal pouch anastomosis. AB - BACKGROUND: Previous studies suggest only minor changes in bile acid metabolism after panproctocolectomy with ileal pouch construction. AIMS/METHODS: To investigate these changes further, we studied cholesterol absorption and serum, biliary and fecal non-cholesterol sterols and lipids in 12 ileal pouch patients and 10 controls. RESULTS: In patients, cholesterol absorption was markedly reduced and was associated with low serum total and LDL cholesterol and LDL triglyceride levels, but surprisingly, cholesterol synthesis, as indicated by sterol-balance data or serum cholesterol precursor levels, was within low normal limits. The high proportions of serum plant sterol to cholesterol, particularly that of campesterol, were not related to cholesterol absorption, but were attributable to a markedly reduced biliary cholesterol secretion. Interestingly, in these patients the fractional absorption of campesterol was normal, whereas that of sitosterol, like cholesterol, was reduced and was positively related to the intestinal influx of cholesterol. The patients' serum cholestanol proportion was normal, but the proportion of the cholestanol formed during intestinal passage was significantly reduced (17.9% vs 65.2% in controls). CONCLUSIONS: Thus ileal pouch patients are characterized by sterol malabsorption, lowered serum total and LDL-cholesterol levels, but unexpectedly without any increase in cholesterol synthesis. The lack of high serum cholestanol, shown earlier frequently in unoperated patients with ulcerative colitis, may indicate reversible cholestasis, a finding deserving further exploration. PMID- 9210619 TI - Heterogeneity of the "oval-cell" response in the hamster liver during cholangiocarcinogenesis following Clonorchis sinensis infection and dimethylnitrosamine treatment. AB - BACKGROUND: Small intraportal "oval" cells which appear in the livers of humans and experimental animals after liver injury, are suspected to be early progenitor cells for both hepatocytes and bile duct cells, as well as cells of origin of hepatocellular and cholangiocellular carcinomas. METHODS: The origin and fate of small "oval" cells expressing different immunohistologic phenotypes and ultrastructural appearance were examined in livers of Syrian hamsters during cholangiocarcinogenesis induced by dimethylnitrosamine and promoted by Clonorchis sinensis infection. RESULTS: Three different "oval" cell types are identified in portal and/or periportal areas: 1) Small periductal cells with abundant heterochromatin and scant cytoplasm that are negative for AFP, CK19, OV-6 and GST p (primitive oval cells); 2) Glycogen-rich cells, positive for AFP, but negative for CK19, OV-6 and GST-p mainly adjacent to ductal plates (hepatocyte-like oval cells); and 3) small cells with desmosomes and basement membrane, containing GST p CK19 and OV-6 but negative for AFP, present in ducts (ductular-like oval cells). It appears that C. sinensis infection stimulates proliferation and differentiation of small ductular or periductal cells (primitive oval cells) into either hepatocyte-like oval cells, which mature into hepatocytes without malignant transformation, or into ductular-like oval cells. CONCLUSIONS: We propose that the ductular-like oval cells are precursors of dysplastic ductular cells that give rise to cholangiocarcinomas after dimethylnitrosamine treatment and conclude that primitive oval cells are bipolar progenitor cells for hepatocytes and biliary cells, and that activation (initiation) of these cells by carcinogen (dimethylnitrosamine), followed by stimulation of proliferation of biliary cells by C. sinensis, promotes primitive oval cells or their progeny (ductular-like oval cells) to transform into cholangiocarcinomas. PMID- 9210620 TI - Woodchuck hepatitis virus-induced carcinoma as a relevant natural model for therapy of human hepatoma. AB - BACKGROUND: Eastern American woodchuck (Marmota monax), naturally infected with woodchuck hepatitis virus, a virus similar to human hepatitis B virus, develops liver cancer with a high prevalence. AIMS: The aim of this work was to assess Marmota monax as a model of human hepatocellular carcinoma, especially to assess new potential adjuvant therapies after surgical resection. METHODS: Forty-four woodchuck hepatitis virus-infected animals were regularly screened by ultrasound examination from the age of 18 months and for a 30-month period. One or more liver tumors were diagnosed in 31 animals (70%). Five of them with multifocal tumor or poor general status were considered unsuitable for surgery. The other 26 were operated on. At laparotomy no tumor was found in three. RESULTS: The 18 liver tumors studied were hepatocellular carcinomas, grossly and microscopically similar to human hepatocellular carcinoma. Peritumoral parenchyma studied in 13 specimens was always non-cirrhotic but adequate staining demonstrated patterns of fibrosis in four cases. Clear evidence of chronic active hepatitis, periportal hepatitis and steatosis were demonstrated in five, seven and one of the 13 specimens, respectively. Tumors were treated by tumorectomy in eight animals, by alcoholization in seven and by laser photocoagulation in one. A simple tumor biopsy was performed in the other seven. Ten animals died postoperatively. All the survivors in the tumorectomy group died from tumor recurrence within 10-18 months after surgery. CONCLUSIONS: It is concluded that woodchuck hepatitis virus induced liver carcinoma is a natural model of human hepatocellular carcinoma with similar pathology and natural history, including early ultrasonic detection and tumor recurrence after resection. Tumor excision is feasible in this animal model, which now provides the basis for assessment of new potential adjuvant therapies for human hepatocellular carcinoma in an attempt to reduce the high recurrence rate after surgical resection in humans. PMID- 9210621 TI - Overexpression of mdr2 gene by peroxisome proliferators in the mouse liver. AB - BACKGROUND: In mice, fibrates induce mdr2 gene expression, and its encoded P glycoprotein in the canalicular domain of hepatocytes, as well as increasing biliary phospholipid output. It is not known whether this effect is restricted to fibrates or is a common property of peroxisome proliferators. AIMS: To test the effect of structurally unrelated peroxisome proliferators on mdr2 gene expression and biliary phospholipid output, and to explore the molecular mechanism(s) of mdr2 gene induction. METHODS: Male CFI mice were fed on a diet supplemented with several peroxisome proliferators: phenoxyacetic acid herbicides, plasticizers, acetylsalicylic acid and partially hydrogenated fish oil. RESULTS: Increased levels of mdr2 mRNAs, assessed by Northern blot analysis, were observed in the liver of mice treated with phenoxyacetic acid herbicides: 2,4,5 trichlorophenoxyacetic acid 570+/-133%, 2,4-dichlorophenoxyacetic acid 233+/-54% (p<0.005); plasticizers: di-(2-ethylhexyl)phthalate 282+/-78%, di (isoheptyl)phthalate 163+/-40%, phthalic acid dinonyl ester 225+/-48% (p<0.01); and partially hydrogenated fish oil 372+/-138% (p<0.005). P-glycoprotein traffic ATPase content increased in the canalicular domain of hepatocyte of mice treated with the herbicide 2,4,5-trichlorophenoxyacetic acid and with partially hydrogenated fish oil (108% and 87%, respectively, p<0.05) as well as biliary phospholipid output (106% and 74%, respectively, p<0.05). In 2,4,5 trichlorophenoxyacetic acid-fed mice we found five-fold increase on mdr2 transcription rate, assessed by nuclear run-off assay. CONCLUSIONS: Peroxisome proliferators induce mdr2 gene, its encoded P-gp in the canalicular domain of hepatocytes and increase biliary phospholipid output. The modulation of mdr2 gene might be part of the pleiotrophic response of peroxisome proliferation in mice liver and seems to be regulated mainly at a transcriptional level. PMID- 9210622 TI - Taurine is an osmolyte in rat liver macrophages (Kupffer cells). AB - BACKGROUND/AIMS: The availability of betaine as an osmolyte was recently shown to interfere strongly with important cell functions of liver macrophages (Kupffer cells), such as eicosanoid and tumor necrosis factor-alpha production or phagocytosis. We therefore investigated whether taurine is also used as an osmolyte by Kupffer cells and whether it is involved in the control of Kupffer cell functions. METHODS/RESULTS: Hyperosmotic (hypoosmotic) exposure of cultured rat liver macrophages (Kupffer cells) for 6-12 h led to an increase (decrease) in the mRNA levels of the taurine transporter (TAUT) and an increase (decrease) in taurine transport into the cells. The hyperosmolarity-induced increase in TAUT mRNA levels was diminished by 37+/-10% upon addition of taurine, but not upon addition of betaine. When Kupffer cells were preloaded with taurine, hypoosmotic exposure led to a rapid efflux of taurine from the cells, which was significantly delayed in the presence of the anion exchanger inhibitor 4,4' diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). Taurine efflux was also stimulated during phagocytosis of Latex particles; however, Latex was without effect on the hyperosmolarity-induced increase of TAUT mRNA levels. Lipopolysaccharide (LPS) led to an induction of cyclooxygenase-2, which was markedly enhanced during hyperosmotic conditions. Taurine diminished the induction of cyclooxygenase-2 and inhibited the LPS/hyperosmolarity-induced stimulation of prostaglandin E2 formation. CONCLUSIONS: The data suggest that, in addition to betaine, taurine also acts as an osmolyte in Kupffer cells, and that taurine availability may be an important modulater of Kupffer cell functions such as eicosanoid synthesis. PMID- 9210623 TI - NO-mediated vasodilation in the rat liver. Role of hepatocytes and liver endothelial cells. AB - BACKGROUND/AIMS: Nitric oxide (NO) is a potent vasodilator. We investigated the mechanisms responsible for this effect in the liver. METHODS: Isolated perfused rat liver and cultures of endothelial sinusoidal cells and hepatocytes were used. RESULTS: L-arginine (10(-3) M) and NO donor Sin-1 (10(-5) M) respectively increased the liver flow by 52% (p<0.01) and 93% (p<0.01) vs controls. The NO synthase inhibitor Nw-nitro-L-arginine (10(-3) M) and the guanylate cyclase inhibitor methylene blue (10(-5) M) respectively decreased the basal liver flow by 26% and 16% (p<0.05) and inhibited the vasodilating effects of L-arginine. L arginine (10(-3) M) increased nitrite concentration in hepatocyte culture (77.25+/-7.40 micromol x l(-1) vs 14.70+/-3.55 micromol x l(-1) in controls; p<0.01) and in liver endothelial cell culture (0.36+/-0.09 micromol x l(-1) vs 0.12+/-0.05 micromol x l(-1) in controls; p<0.05). Nw-nitro-L-arginine inhibited the basal production and abolished the L-arginine-induced production of nitrites both in hepatocyte and in liver endothelial cell cultures. The concentration of nitrites in the hepatocyte supernatant rose from 14.70+/-3.55 micromol x l(-1) to 150.50+/-45.55 micromol x l(-1) in the presence of a combination of interleukin 1beta, TNF alpha and interferon gamma. CONCLUSIONS: Under basal conditions, NO regulates the vascular tone of liver circulation. Both liver endothelial cells and hepatocytes can be implicated. NO production by hepatocytes may increase during inflammation. PMID- 9210624 TI - Urinary excretion of the collagen cross-link pyridinoline increases during liver fibrogenesis. AB - BACKGROUND/AIMS: Pyridinoline, a specific cross-link of mature collagen, increases in liver during fibrogenesis and its hepatic level is related to the degree of reversibility of the fibrotic process. Since pyridinoline is excreted in urine, we have investigated the relationship between its urinary level and liver fibrogenesis in a model of mild and reversible liver fibrosis, murine schistosomiasis. METHODS: Pyridinoline was measured by HPLC in urine and in liver of Schistosoma mansoni-infected mice during the acute and the chronic phases of the infection. Collagen deposition was measured colorimetrically. Both the isolated granulomas and the surrounding liver parenchyma were analyzed. RESULTS: In infected mice, pyridinoline increased mainly in the isolated granulomas, corresponding to the fibrotic lesions, and slightly in the surrounding parenchyma. The urinary excretion of pyridinoline increased during liver fibrogenesis and was correlated to the duration of infection (r=0.81) and to the collagen content of granulomas (r=0.81). The treatment of infected mice by praziquantel, an antiparasitic drug, did not lead to significant changes in liver collagen cross-linking by pyridinoline either in granulomas or in parenchyma. The major effect of the drug was targeted at the collagen content of parenchyma, which decreased by 50%, 18 weeks after treatment. The urinary level of pyridinoline of treated mice was negatively correlated to the length of the treatment follow-up (r=-0.76). CONCLUSIONS: The measurement of the urinary excretion of pyridinoline could be helpful to monitor the remodeling of liver extracellular matrix occurring in fibrogenesis and the effect of chemotherapy. PMID- 9210625 TI - Effects of simvastatin, pentoxifylline and spironolactone on hepatic fibrosis and portal hypertension in rats with bile duct ligation. AB - AIMS/METHODS: Our aim was to study the antifibrotic and hemodynamic effects of simvastatin (SMV), pentoxifylline (PTX) and spironolactone (SPN), three drugs which may have antifibrotic and/or portal hypotensive properties, in a model of hepatic fibrosis and portal hypertension induced in rats by bile duct ligation. A blind study was performed in five groups of 53 Sprague-Dawley rats: sham, placebo (PL), SMV (2.5 mg x kg(-1) x J(-1)), PTX (50 mg x kg(-1) x J(-1)) and SPN (100 mg x kg(-1) x J(-1)). Drugs were administered by daily gavage over a 4-week period as soon as bile duct ligation was performed. At day 28, the following parameters were evaluated: area of hepatic fibrosis by image analysis after staining collagen with picrosirius and plasma concentrations of hyaluronate, splanchnic and systemic hemodynamics (radiolabeled microspheres). RESULTS: Portal venous pressure (PL: 15.5+/-1.5, SMV: 15.8+/-2.5, PTX: 15.9+/-1.8, SPN: 13.5+/-2.1 mmHg, p<0.05) and porto-systemic shunts (PL: 30+/-31, SMV: 18+/-27, PTX: 25+/-24, SPN: 5+/-4%, p<0.05) were significantly reduced in the SPN group; other hemodynamic parameters were not significantly altered. There was a significant correlation between portosystemic shunts and portal pressure (r(s)=0.47, p<0.01). The area of fibrosis was not significantly different among the four groups of bile duct ligated rats (PL: 8.7+/-3.9, SMV: 7.1+/-3.6, PTX: 7.8+/-2.7, SPN: 6.6+/-3.3%) but was higher than in sham rats (1.5+/-0.5%, p<0.001). Hyaluronate was significantly higher in bile duct ligated rats (from 374+/-162 to 420+/-131 microg/l, among the four groups) than in sham rats (52+/-16 microg/l, p<0.0001). CONCLUSIONS: In this model, none of the drugs prevented hepatic fibrosis. On the other hand, spironolactone decreased portal pressure and prevented porto-systemic shunts. Therefore, this drug may have beneficial effects in patients with early portal hypertension. PMID- 9210626 TI - Intestinal bacterial overgrowth and bacterial translocation in cirrhotic rats with ascites. AB - BACKGROUND/AIMS: Translocation of indigenous bacteria from the gut lumen of cirrhotic animals to mesenteric lymph nodes appears to be an important step in the pathogenesis of spontaneous bacterial peritonitis. However, the sequence of events leading to translocation remains unclear. One of the most predictable risk factors for translocation is overgrowth of gut bacterial flora. The present study was designed to compare the intestinal aerobic bacterial flora of cecal stools at the time of sacrifice between cirrhotic and normal rats and to evaluate the role of intestinal aerobic bacterial overgrowth in bacterial translocation in cirrhotic rats. METHODS: Thirty-five male Sprague-Dawley rats with carbon tetrachloride-induced cirrhosis and ascites and 10 normal rats were included in this study. Cirrhotic rats were sacrificed when ill and samples of ascitic fluid, mesenteric lymph nodes and cecal stool were taken for detecting quantitatively aerobic bacteria. RESULTS: Total intestinal aerobic bacterial count in cecal stool at the time of sacrifice was significantly increased in cirrhotic rats with bacterial translocation with or without spontaneous bacterial peritonitis compared to cirrhotic rats without bacterial translocation (p<0.001 and p<0.001, respectively) and to normal rats (p<0.001 and p<0.001, respectively). Of the 42 species of bacteria translocating to the mesenteric lymph nodes, 41 (97.6%) were found in supranormal numbers in the stool at the time of sacrifice. CONCLUSIONS: Carbon tetrachloride-induced cirrhotic rats with bacterial translocation have increased total intestinal aerobic bacteria count, and intestinal bacterial overgrowth appears to play an important role in bacterial translocation in this experimental model of cirrhosis in rats. PMID- 9210627 TI - In vitro evaluation of a novel bioreactor based on an integral oxygenator and a spirally wound nonwoven polyester matrix for hepatocyte culture as small aggregates. AB - BACKGROUND/AIMS: The development of custom-made bioreactors for use as a bioartificial liver (BAL) is considered to be one of the last challenges on the road to successful temporary extracorporeal liver support therapy. We devised a novel bioreactor (patent pending) which allows individual perfusion of high density cultured hepatocytes with low diffusional gradients, thereby more closely resembling the conditions in the intact liver lobuli. METHODS: The bioreactor consists of a spirally wound nonwoven polyester matrix, i.e. a sheet-shaped, three-dimensional framework for hepatocyte immobilization and aggregation, and of integrated hydrophobic hollow-fiber membranes for decentralized oxygen supply and CO2 removal. Medium (plasma in vivo) was perfused through the extrafiber space and therefore in direct hepatocyte contact. Various parameters were assessed over a period of 4 days including galactose elimination, urea synthesis, lidocaine elimination, lactate/pyruvate ratios, amino acid metabolism, pH, the last day being reserved exclusively for determination of protein secretion. RESULTS: Microscopic examination of the hepatocytes revealed cytoarchitectural characteristics as found in vivo. The biochemical performance of the bioreactor remained stable over the investigated period. The urea synthesizing capacity of hepatocytes in the bioreactor was twice that of hepatocytes in monolayer cultures. Flow sensitive magnetic resonance imaging (MRI) revealed that the bioreactor construction ensured medium flow through all parts of the device irrespective of its size. CONCLUSIONS: The novel bioreactor showed encouraging efficiency. The device is easy to manufacture with scale-up to the liver mass required for possible short-term support of patients in hepatic failure. PMID- 9210628 TI - Lamivudine resistance in immunocompetent chronic hepatitis B. Incidence and patterns. AB - BACKGROUND: Lamivudine is a non-toxic, potent inhibitor of hepatitis B virus replication. Recently, hepatitis B virus resistance to lamivudine has been described in patients using immunosuppressive drugs after liver transplantation. METHODS: From our cohort of 81 consecutive patients treated with lamivudine, we selected all immunocompetent patients who received lamivudine monotherapy for a period over 26 weeks (n=14). RESULTS: Lamivudine resistance with the characteristic mutation in the YMDD motif was observed in four patients (actuarial cumulative incidence: 39%). Two patterns of viral resistance were observed: incomplete response (n=2) and viral breakthrough (n=2). CONCLUSIONS: The observed high frequency of lamivudine resistance may have implications for the concept of long-term virus-suppressive therapy of chronic hepatitis B by lamivudine monotherapy. PMID- 9210629 TI - Establishment of a novel radioligand assay using eukaryotically expressed cytochrome P4502D6 for the measurement of liver kidney microsomal type 1 antibody in patients with autoimmune hepatitis and hepatitis C virus infection. AB - BACKGROUND/AIMS: Liver kidney microsomal type 1 antibody (LKM1) is the diagnostic marker of autoimmune hepatitis (AIH) type 2 and is also found in patients with hepatitis C virus (HCV) infection. Cytochrome P4502D6 (CYP2D6) is the documented target antigen of LKM1 in AIH, but not in HCV infection. To compare the reactivity in the two conditions, we established a radioligand assay using eukaryotically expressed CYP2D6 as target. METHODS: A 1.2-kb human CYP2D6 cDNA was isolated from a human liver cDNA library and subcloned into an in vitro transcription vector pSP64 Poly(A). Recombinant CYP2D6 was then produced by in vitro transcription/translation, metabolically labelled with 35S methionine and used in the immunoprecipitation assay. Antibodies that bound radiolabelled CYP2D6 were immunoprecipitated and their levels assessed as cpm. Sera from 50 LKM1 positive patients (26 with AIH; 24 with HCV infection), 128 LKM1-negative patients and 57 normal controls were tested. RESULTS: Reactivity to 35S labelled CYP2D6 was observed in all LKM1-positive sera from patients with AIH and HCV infection, but in none of the controls. The cpm in both conditions were significantly higher than in normal controls (p<0.0001), and were correlated with the immunofluorescence titres of LKM1 (r 0.87, p<0.001 and r=0.64, p<0.001 for AIH and HCV infection, respectively). Reactivity to 35S labelled CYP2D6 was inhibited by addition of an excess of eukaryotically expressed CYP2D6. CONCLUSIONS: CYP2D6 is a major target antigen of both AIH and HCV infection. The novel radioligand assay is highly sensitive and specific. PMID- 9210630 TI - Severe alpha1-antitrypsin deficiency (PiZ homozygosity) with membranoproliferative glomerulonephritis and nephrotic syndrome, reversible after orthotopic liver transplantation. AB - BACKGROUND/AIMS: Nephropathy associated with alpha1-antitrypsin deficiency is assumed to be an unusual entity. We describe the case of a 23-year-old woman with severe alpha1-antitrypsin (PiZ homozygosity) deficiency who developed hepatic cirrhosis in childhood, and glomerulonephritis and nephrotic syndrome in adult life. METHODS/RESULTS: A renal biopsy was consistent with membranoproliferative glomerulonephritis. An immunofluorescence study revealed the presence of alpha1 antitrypsin (PiZ) in the subendothelial region of the glomerular basement membrane. The renal disease was reversible after orthotopic liver transplantation. CONCLUSIONS: The presence of abnormal PiZ protein in the subendothelial region of the glomerular basement membrane may suggest a possible role for this protein in the pathogenesis of glomerulonephritis. The case should add impetus to the search for alpha1-antitrypsin deficiency in any patient presenting with combined liver and renal disease, in the absence of evidence of hepato-renal syndrome, and illustrates that liver transplantation alone may reverse the nephropathy associated with alpha1-antitrypsin deficiency. PMID- 9210631 TI - Resolution of hydatid liver cyst by spontaneous rupture into the biliary tract. AB - Among the complications of hydatid liver disease, spontaneous cyst rupture into the biliary tract is unusual, occurring in 3.2-17% of cases. Its endoscopic management has been reported rarely, and corresponding complete photodocumentation is unique. Such a case is described and comprehensively illustrated in a 48-year-old immunocompromised man, presenting with upper abdominal pain, obstructive jaundice, and fever. Impaction of hydatid material into the common bile duct and the papilla of Vater was relieved endoscopically, and the patient was consecutively treated with two courses of mebendazole. This management resulted in complete clinical resolution of hepatic hydatosis after 8 months of follow-up. Complications of overt cyst perforation may be allergic, obstructive, secondary infectious, or metastatic. Ultrasound and computed tomography are complementary tools for diagnosis of hepatic echinococcosis, with endoscopic retrograde cholangiography being the "gold standard" in confirming rupture into the biliary system. Laboratory results are usually non-specific. While surgical excision is the treatment of choice, selected patients may primarily be managed endoscopically, followed by anthelminthic therapy. PMID- 9210632 TI - Images in hepatology. Cruveilhier-Baumgarten syndrome. PMID- 9210633 TI - Prognostic models in chronic liver disease: validity, usefulness and future role. PMID- 9210634 TI - Evidence in favour of high infection rate with hepatitis E virus among young children in India. PMID- 9210635 TI - Decreased serum alfa2-HS-glycoprotein concentration in patients with primary biliary cirrhosis. PMID- 9210636 TI - Biliary obstruction caused by portal cavernoma in a patient with laterality sequence. PMID- 9210637 TI - Successful liver transplantation using a polycystic donor liver. PMID- 9210638 TI - Gaseous pathway in venous oxygen persufflation of the liver. PMID- 9210639 TI - Identification of a reactive cysteine in the flavin-binding domain of Rhodotorula gracilis D-amino acid oxidase. AB - The holoenzyme form of Rhodotorula gracilis D-amino acid oxidase, an 80-kDa homodimer, reacted only to a limited extent with general thiol reagents (2,2' dithiodipyridine, 5,5'-dithiobis(2-nitrobenzoic acid), and N-[7-(dimethylamino)-4 methylcoumarinyl]maleimide) (60% residual activity), whereas the monomeric apoprotein was completely inactivated and denatured by these reagents. To investigate the presence of thiol residue(s) in the active site of the enzyme, the apoprotein was reconstituted with the 8-(methylsulfonyl)-FAD chemical affinity probe. Competitive inhibition between this analogue and FAD for apoprotein binding was observed. The covalent attachment of the flavin analogue to the apoprotein was complete after approximately 20 h of incubation and the flavinylated enzyme, containing 8-(cysteinyl)-FAD, was monomeric and inactive. After HPLC isolation of the flavin-labeled tryptic peptides, Cys208 was identified as the only cysteine to react with the FAD analogue. These results show that a single cysteine of R. gracilis D-amino acid oxidase reacts with the flavin analogue and that this is located near or at the FAD-binding domain. PMID- 9210640 TI - Fusion of sperm with prostasomes: effects on membrane fluidity. AB - Prostasomes are membranous vesicles (150-200 nm diameter) present in human semen. They are secreted by the prostate gland and contain large amounts of cholesterol, sphingomyelin, and Ca2+. In addition, some of their proteins are enzymes. Prostasomes enhance the motility of ejaculated sperm and are involved in a number of biological functions. In this work, we study the fusion of prostasomes to sperm by determining the relief of octadecylrhodamine self-quenching and the fluidity of membranes by measuring the fluorescence anisotropy of diphenylhexatriene. We present the following findings: (a) the contact of sperm cells with prostasomes at slightly acidic pH causes the fusion of the membranes; (b) the amount of transferred lipid depends on the prostasome/sperm ratio; (c) the fluidity of sperm is much higher than that of prostasomes; (d) the fusion changes some properties of sperm cells, such as fluidity, which decreases greatly; and (e) the extent of fluidity variations depends on the prostasome to sperm ratio. We propose that the H(+)-dependent fusion of prostasomes to sperm may have physiological consequences. In fact, this process can modify the lipid and protein pattern of sperm plasma membranes. PMID- 9210641 TI - Antioxidant reactions of all-trans retinol in phospholipid bilayers: effect of oxygen partial pressure, radical fluxes, and retinol concentration. AB - Lipoperoxyl radical-scavenging activity of retinol in unilamellar soybean phosphatidylcholine liposomes was studied under a variety of conditions to appreciate to what extend retinol may be considered an effective antioxidant. Peroxidation, initiated by 2 mM 2,2'-azobis(amidino-propane)hydrochloride (AAPH), was carried out at 160 torr O2 or at 15 torr O2, in the absence or in the presence of 10 to 40 mM retinol. As evaluated by the length of the inhibition periods, t(inh), and by the ratio between the inhibition and propagation rate, R(inh)/R(p), the antioxidant activity of retinol was higher at 15 torr O2 than at 160 torr O2. The consumption rate of retinol was markedly faster at 160 torr O2 than at 15 torr O2 and increased with the increase of retinol concentration under both oxygen tensions. When liposome peroxidation was carried out under N2, retinol consumption was independent of retinol concentration. Peroxyl radicals oxidize retinol to 5,6-retinol epoxide. The ratio between 5,6-epoxide formed and the retinol consumed was markedly higher at 15 torr O2 than under air and decreased with the increased retinol concentrations. When butylated hydroxytoluene was included into the liposomal suspension, most of the consumed retinol was converted into 5,6-epoxide. Liposomes were incubated at 15 torr O2, in the presence of 0.5 to 10 mM AAPH. The antioxidant effectiveness of 40 mM retinol, as measured by the R(inh)/R(p) ratio, increased with the increase of the radical fluxes. The results suggest, besides radical trapping, that a major consumption of retinol during lipid oxidation occurs through self-oxidation reactions, which are concentration- and oxygen-dependent. A decreased self oxidation makes retinol a better lipoperoxyl radical scavenger at low, rather than at high partial pressure of oxygen. However, when self-oxidation of retinol is prevented, only a minor fraction of the antioxidant is allowed to effectively act as a radical scavenger, suggesting that the radical-trapping reactions are rate-limiting for the antioxidant process. Peroxyl radical concentration, by shifting the route of the retinol activity toward radical scavenging, brings about an increasingly more efficient radical trapping. It is concluded that all trans retinol behaves as a more effective antioxidant at low oxygen partial pressure, low retinol concentrations, and high radical flux. PMID- 9210642 TI - Synthesis of dolichol in a polyprenol reductase mutant is restored by elevation of cis-prenyl transferase activity. AB - CHB11-1-3 is a glycosylation mutant of Chinese hamster ovary (CHO) cells, isolated by screening mutagenized cells for those with decreased intracellular lysosomal enzyme activity [C. W. Hall et al. (1986) Mol. Cell. Biochem. 72, 35 45]. CHB11-1-3 synthesizes the lipid polyprenol, the metabolic precursor of dolichol, rather than dolichol, indicating a defect in polyprenol reductase. This defect was demonstrated previously in Lec9 CHO mutants, and cell fusion experiments confirmed that CHB11-1-3 is a member of this complementation group. A revertant of CHB11-1-3, CHBREV, isolated for its ability to grow at 39 degrees C, synthesizes dolichol at near-normal levels. CHBREV is probably a second-site revertant, because it synthesizes three to four times as much polyprenol as CHB11 1-3 and exhibits a similar elevation in the specific activity of cis-prenyl transferase. This higher activity appears to reflect an increase in enzyme molecules rather than the presence of an activator or absence of an inhibitor. These results suggest that CHB11-1-3 is a "K(m)" mutant, because synthesis of higher amounts of the substrate of polyprenol reductase obviates the defect. PMID- 9210643 TI - Identification of rat liver glucose-3-phosphatase as an inositol monophosphatase inhibited by lithium. AB - Glucose-3-phosphatase (Glc3Pase) from rat liver has been purified 780-fold with a 4% recovery. The substrate specificity of the purified enzyme agreed with that of inositol monophosphatase (EC 3.1.3.25). D-Glucose 3-phosphate (D-Glc(3)P; K(m) = 200 microM) was hydrolyzed with an efficiency similar to DL-myo-inositol 1 monophosphate (DL-Ins(1)P; K(m) = 80 microM), since the ratio V(max)/K(m) was similar for both substrates. Purification data, coelution of activities, thermal inactivation curves, optimal pH, bivalent cation requirements, inhibition by Li+, molecular weight, and isoelectric pH comparisons supported that the hydrolysis of D-Glc(3)P and DL-Ins(1)P was catalyzed by a unique phosphohydrolase identified as a hepatic form of the lithium-sensitive inositol monophosphatase. That the hydrolysis of D-Glc(3)P is a genuine feature of inositol monophosphatases was confirmed because the enzyme purified from bovine brain showed also Glc3Pase activity, and inspection of published 3D models of inositol monophosphatase complexes with D(L)-Ins(1)P or D(L)-Ins(4)P indicated that beta(alpha)-D-Glc(3)P in a pyranose conformation with all (but one) the hydroxy groups in equatorial orientation would fit in the active site as other good substrates do. The results of this work are suggestive of possible relationships between inositol and sugar 3-phosphate metabolism. PMID- 9210644 TI - Sequence and biological activity of catrocollastatin-C: a disintegrin like/cysteine-rich two-domain protein from Crotalus atrox venom. AB - In this study we report on the isolation and biological characterization of a 23.6-kDa protein from the venom of the western diamondback rattlesnake, Crotalus atrox. Primary structural analysis shows the protein to be composed of a spacer/disintegrin-like domain and a cysteine-rich domain. The sequence is identical to the same carboxy-terminal domains found in the C. atrox metalloproteinase, catrocollastatin, and hence we termed the protein catrocollastatin-C. We estimate that catrocollastatin-C represents at least 0.5% of the total protein in C. atrox venom. The protein is an inhibitor of collagen stimulated but not ADP-stimulated platelet aggregation. Reduction and alkylation of catrocollastatin-C causes a loss of platelet aggregation inhibitor activity. A synthetic, cyclic peptide designed from the catrocollastatin-C disintegrin-like domain has potent platelet aggregation inhibitory activity. This suggests that the corresponding region in the disintegrin-like domain of the protein is at least partially responsible for the inhibition of platelet aggregation previously reported for the protein. These studies underscore the biochemical and functional complexity of crotalid snake venoms due to differential proteolytic processing of precursor proteins and how the processed precursor fragments may contribute to the observed pathological effects of the venom. PMID- 9210646 TI - Identification of MAPKAPK homolog (MAPKAPK-4) as a myosin II regulatory light chain kinase in sea urchin egg extracts. AB - We identified and cloned a homolog of mammalian mitogen-activated protein kinase activated protein kinase (MAPKAPK)-2 and -3 from sea urchin, Hemicentrotus pulcherrimus. The obtained cDNA clone was composed of 350 amino acid residues which contain MAPK phosphorylation sites and the bipartite nuclear localization signal sites in its C-terminal domain. The clone showed 65.4 and 66.7% amino acid residue identity to human MAPKAPK-2 and -3, respectively. Phylogenetic analysis revealed that the homolog can be classified into a distinct group of MAPKAPK and, therefore, the identified homolog was designated as MAPKAPK-4. Biochemical characterization was performed using recombinant glutathione S-transferase (GST) MAPKAPK-4 fusion protein. The protein kinase activity of GST-MAPKAPK-4 was activated by MAPK and this enabled the kinase to phosphorylate both glycogen synthase N-terminal peptide and the regulatory light chain of myosin II in vitro. Northern blot analysis showed that MAPKAPK-4 was expressed throughout the development of sea urchin embryos. These observations suggest that MAPKAPK-4 may play an important role in the regulation of myosin II activity during the development of sea urchin. PMID- 9210645 TI - Phospholipid peroxidation induces cytosolic phospholipase A2 activity: membrane effects versus enzyme phosphorylation. AB - Cytosolic phospholipase A2 (cPLA2) is a signal-responsive enzyme that is highly selective to the nature of phospholipid substrates. A mechanism for cPLA2 activity regulation through a signal transduction pathway has been proposed and this signaling appears to be influenced by oxidants. Oxidant-mediated signaling of PLA2 may serve as an alternative mechanism for enzyme regulation; however, the manner of regulation has yet to be delineated. In this report we demonstrate that there is a direct effect of membrane oxidation on cPLA2 phosphorylation and activity. A simple in vitro system consisting of purified cPLA2 and phospholipid vesicles was used to facilitate protein kinase C (PKC) activity and provide substrates for cPLA2. Using these vesicles we found that the activity of cPLA2 was enhanced twofold when the vesicles contained as little as 5 mol% phosphatidylcholine hydroperoxides (PLPCOOH). The order of hydrolytic preference for fatty acyl species was 20:4 > 18:2 > 18:1 > 16:0, and the presence of PLPCOOH stimulated hydrolysis largely of phosphatidylcholine containing 20:4. The Ca2+ concentrations required for stimulated hydrolytic activity were also twofold lower for oxidized compared to unoxidized vesicles. Using phospholipid micelles as substrates, PKC-mediated phosphorylation of cPLA2 increased hydrolytic activity 71% compared to preparations lacking PKC. Using phospholipid vesicles as substrates, PKC-mediated phosphorylation resulted in an 85% increase in cPLA2 activity compared to preparations without PKC. PKC-mediated phosphorylation of cPLA2, therefore, stimulates catalytic activity toward membrane phospholipids and the extent of activation is enhanced directly by peroxidation of membrane phospholipids and involves a peroxide-induced stimulation of cPLA2 phosphorylation. PMID- 9210647 TI - Chemical modification of deoxycytidine at different sites yields adducts of different stabilities: characterization of N3- and O2-deoxycytidine and N3 deoxyuridine adducts of butadiene monoxide. AB - Eight adducts were characterized from the reaction of deoxycytidine with the chemical carcinogen, butadiene monoxide (BM). They were identified as diastereomeric pairs of N3-(2-hydroxy-3-buten-1-yl)deoxycytidine, N3-(2-hydroxy-3 buten-1-yl)deoxyuridine, N3-(1-hydroxy-3-buten-2-yl)deoxyuridine, and O2-(2 hydroxy-3-buten-1-yl)deoxycytidine based on UV spectra, 1H NMR, FAB/MS, and stability studies. The N3-(2-hydroxy-3-buten-1-yl)deoxycytidine adducts were unstable at pH 7.4, 37 degrees C, and deaminated to the corresponding N3 deoxyuridine adducts with half-lives of 2.3 and 2.5 h. The N3-(1-hydroxy-3-buten 2-yl)deoxycytidine diastereomers were not detected, apparently because of faster rates of deamination compared to the N3-(2-hydroxy-3-buten-1-yl)deoxycytidine adducts. The corresponding four N3-deoxyuridine adducts were stable for up to 168 h. The O2-deoxycytidine adducts were unstable and decomposed with a half-life of 11 h. The N3-(2-hydroxy-3-buten-1-yl)deoxycytidine adducts were initially the major adducts formed upon reaction of deoxycytidine with BM at 37 degrees C in phosphate buffer (pH 7.4), but the corresponding N3-deoxyuridine adducts showed a lag in formation due to the time needed for deamination. The N3-(1-hydroxy-3 buten-2-yl)deoxyuridine and O2-deoxycytidine adducts had linear formation rates, but were formed to a lesser extent. Heating the reaction mixture at 80 degrees C for 1 h converted all N3-deoxycytidine adducts to the stable N3-deoxyuridine adducts. Incubation of deoxycytidine with an excess of BM at pH 7.4, 37 degrees C, followed by the extraction and heating steps allowed calculation of the pseudo first-order kinetic rate constants for the four uridine adducts. If the heating step was eliminated, then the pseudo-first-order kinetic rate constants could be calculated for the N3-(2-hydroxy-3-buten-1-yl)deoxycytidine and O2-(2-hydroxy-3 buten-1-yl)deoxycytidine adducts. The rate constants for N3-(2-hydroxy-3-buten1 yl)deoxycytidine and the corresponding N3-(2-hydroxy-3-buten-1-yl)deoxyuridine were five- to sixfold the rate constants for the N3-(1-hydroxy-3-buten-2 yl)deoxyuridine and O2-(2-hydroxy-3-buten-1-yl)deoxycytidine adducts. Thus, the results show that the reaction of deoxycytidine with BM yields adducts at different sites with different rates of formation and stabilities. Understanding the chemical interactions of deoxycytidine with BM and the stability of the various adducts may contribute to a better understanding of the molecular mechanisms of mutagenesis and carcinogenesis of BM and the development of useful biomarkers of exposure. PMID- 9210648 TI - Structure of the O-linked oligosaccharides from a major thyroid cell surface glycoprotein. AB - A major glycoprotein at the surface of calf thyroid cells, GP-3 (M(r) 20,000), contains the I-antigenic activity of calf thyroid which has been attributed to its poly-N-acetyllactosamine N-linked saccharide chains (Edge, A. S. B., and Spiro, R. G., J. Biol. Chem. 260, 15332-15338, 1985). The present study demonstrated that alkaline borohydride treatment of GP-3 results in the release of five neutral and five acidic saccharides that were found to represent over 30% of the saccharides of this carbohydrate-rich glycoprotein. Three of the oligosaccharides contained terminal alpha1 --> 3-linked galactose residues which accounted for their affinity toward Bandeiraea simplicifolia I lectin. The saccharides could be grouped into several distinct categories on the basis of their internal sequence. A novel tetrasaccharide in GP-3 was shown to have the structure: Gal alpha1 --> 3Gal beta1 --> 6(Gal beta1 --> 3)GalNAcH2. An unsubstituted N-acetylgalactosamine unit and a Gal beta1 --> 3GalNAc disaccharide were prominent O-linked constituents, with the disaccharide serving as a core unit for the attachment of sialic acid residues to form tetra- and trisaccharides. A branched core structure, Gal beta1 --> 3(GlcNAcbeta1 --> 6)GalNAcH2 was shared by 4 of the 10 saccharides, the most complete of which was assigned the sequence NeuAc alpha2 --> 3Gal beta1 --> 3(Gal alpha1 --> 3Gal beta1 --> 4GlcNAc beta1 --> 6)GalNAcH2. PMID- 9210650 TI - Pattern and spacing of basic amino acids in heparin binding sites. AB - Glycosaminoglycan (GAG)-protein interactions regulate a myriad of physiologic and pathologic processes, yet an understanding of how these molecules interact is lacking. The role of the pattern and spacing of basic amino acids (arginine (R) and lysine (K)) in heparin binding sites was investigated using peptide analogs as well as by examining known heparin binding sites. Peptides having the general structure R(n)W (n = 3-9, where tyrosine (W) was added for peptide detection) were synthesized and their interaction with heparin was determined by isothermal titration calorimetry. Binding affinity increased with increasing number of R residues. A 9-mer of R (R9W) bound as tightly to heparin as acidic fibroblast growth factor under physiologic conditions. Despite their high affinity for heparin, long stretches of basic amino acids are uncommon in heparin binding proteins. Known heparin binding sites most commonly contain single isolated basic amino acids separated by one nonbasic amino acid. Peptides having the structure, H3CCONH-GRRG(m)RRG(5-m)-CONH2 (denoted as the RRG(m)RR peptide series) and H3CCONH-GRRRG(m)RG(5-m)-CONH2 (denoted as the RRRG(m)R peptide series), where m = 0-5, were synthesized to test the hypothesis that the spacing of basic amino acids in heparin binding sites is optimally arranged to interact with different GAGs. The peptides, in both the -RRG(m)RR- and -RRRG(m)R- peptide series, when m = 0, bound most tightly with heparin, as measured by affinity chromatography. In contrast, the -RRG(m)RR-peptide series interacted most tightly with heparan sulfate when m = 0 or 1, whereas the -RRRG(m)R- peptide series bound tightest when m = 3. These results are consistent with our understanding of heparin and heparan sulfate structure. A highly sulfated GAG, such as heparin, interacts most tightly with peptides (or peptide sequences within proteins) containing a complementary binding site of high positive charge density. Heparan sulfate, having fewer and more highly spaced negatively charged groups, interacts most tightly with a complementary site on a peptide (or peptide sequences with proteins) that has more widely spaced cationic residues. PMID- 9210649 TI - Isolation, characterization, and functional studies of rat liver iron regulatory protein 1. AB - Ferritin mRNAs are translationally regulated by the binding of either of two cytosolic proteins, iron regulatory protein 1 (IRP1) or IRP2, to the iron responsive element (IRE) located in their 5' untranslated region (UTR). Rat liver IRP1 was purified by anion exchange, gel filtration, and affinity chromatography using a concatemerized version of the IRE. Two bands with M(r) of 95,000 and 100,000 were observed by reducing SDS-PAGE. A single protein was responsible for both bands since: (1) [32P]IRE RNA specifically cross-linked to both components; (2) alkylation with iodoacetamide resulted in formation of a single species with M(r) of 95,000; and (3) they possessed identical peptide patterns after digestion with cyanogen bromide. The N-terminal sequence of rat liver IRP1 was MKNPFAHLAEPLDPAQPGKKFNLNKLEDSRYGRLPFXIRVLLEAAV which is identical to the sequence deduced from the cDNA. Rat liver IRP1 has an amino acid composition similar to that of bovine liver caconitase. Several species of IRP1 were observed by two dimensional gel electrophoresis with pIs ranging from 7.5 to 8.0. Rat liver IRP1 bound the IRE with high affinity (K(D) = 0.04 nM) and repressed translation of ferritin mRNA in vitro. IRP1 bound 100-fold less well to an IRE variant and failed to significantly repress translation of a ferritin mRNA containing the mutated IRE. We conclude that decreases in the affinity of interaction between IRP1 and the IRE, of a magnitude similar to that observed when the binding protein in converted to c-aconitase, are sufficient to significantly enhance translation of ferritin mRNA in vitro. PMID- 9210651 TI - Alternative splicing of CYP2D mRNA in human breast tissue. AB - The human cytochrome P450 (CYP) 2D subfamily comprises the CYP2D6 gene and four pseudogenes, CYP2D7P1 and 2 and CYP2D8P1 and 2. The CYP2D6 gene product is a prominent drug-metabolizing enzyme, which is probably constitutive and has no known inducing agents. Alternative splicing of the pre-mRNAs of these genes has been detected in human liver and breast tissue. RNA-PCR, competitive RNA-PCR, Southern blotting, cDNA sequencing, and gene-specific PCR have been used to fully characterize the alternatively spliced forms of CYP2D mRNA in human breast tissue in the region of exon 5 to 8. Such alternative splicing could regulate the expression of CYP2D6 protein. A full-length mRNA (exons 5 to 8), and variants c (exon 6 deleted), b' (3' portion of exon 6 deleted), e (3' portion of exon 6 deleted, 3' 57-bp portion of intron 6 included), d (3' 57-bp portion of intron 6 included), and b (intron 6 included) were characterized and quantitated. Variant c was derived from CYP2D6, variants d, e, and b were from CYP2D7P, and variant b' and full-length mRNA were derived from both CYP2D6 and 2D7P. Full-length mRNA was a minor form in human breast tissue where variants b' and c predominated. Human breast tumor MCF-7 cells had CYP2D mRNA splice variant patterns similar to those of human breast tissue, while human liver tumor HepG2 cells had wild-type mRNA predominating. These results suggest that CYP2D6 could be regulated tissue specifically using tissue-specific alternative mRNA splicing. PMID- 9210652 TI - The mechanism of inactivation of human placental S-adenosylhomocysteine hydrolase by (E)-4',5'-didehydro-5'-methoxyadenosine and adenosine 5'-carboxaldehyde oxime. AB - The mechanisms by which (E)-4',5'-didehydro-5'-methoxyadenosine (DMOA) and adenosine 5'-carboxaldehyde oxime (ACAO) inactivate S-adenosylhomocysteine (AdoHcy) hydrolase were elucidated in this study. Their inhibitory activities toward AdoHcy hydrolase were found to be time- and concentration-dependent, and DMOA and ACAO had K(i) and k2 values of 3.0 microM and 0.10 min(-1) and 0.67 microM and 0.16 min(-1), respectively. The inactivation of AdoHcy hydrolase by DMOA (and ACAO) occurs concomitantly with the reduction of the enzyme-bound NAD+ to NADH. The rates of enzyme inactivation correspond to the rates of NADH formation. Incubation of both DMOA and ACAO with the NAD+ form of AdoHcy hydrolase resulted in formation of 3'-ketoadenosine (3'-keto-Ado) 5' carboxaldehyde and its 4'-epimer. Incubation of DMOA and ACAO with the apo form of the enzyme afforded adenosine (Ado) 5'-carboxaldehyde and its 4'-epimer. These results show that DMOA and ACAO are "proinhibitors" of the enzyme. They are first converted to the inhibitors (Ado 5'-carboxaldehyde and its 4'-epimer) in the active site of the enzyme; these inhibitors then inactivate the enzyme by a type I mechanism. The results from this study demonstrated that this is a common mechanism by which 4',5'-didehydroadenosine analogs, serving as substrates of both the 5'-hydrolytic activity and the 3'-oxidative activity of the enzyme, inactivate AdoHcy hydrolase. The results also provide further evidence supporting the hypothesis that AdoHcy hydrolase possesses a 5'-hydrolytic activity independent of the 3'-oxidation activity. PMID- 9210653 TI - 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors unmask cryptic regulatory mechanisms. AB - The possibility that potent inhibitors of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase may alter the mechanisms by which dietary cholesterol and farnesol regulate this gene was investigated by comparing the regulatory responses of rats maintained on diets with or without 0.04% Lovastatin supplementation to dietary cholesterol. It was found that the rate of hepatic HMG CoA reductase transcription was significantly decreased by dietary cholesterol in animals fed Lovastatin-supplemented diets, whereas animals maintained on a normal chow diet showed no decrease in the rate of transcription. The levels of reductase mRNA were decreased to about 10% of controls in Lovastatin-supplemented animals in response to dietary cholesterol but not affected in nonsupplemented animals. Administration of farnesol, reputed to be the nonsterol regulator of reductase, to rats maintained on a diet containing Lovastatin decreased hepatic HMG-CoA reductase protein by 30% and the half-life of reductase immunoreactive protein to 4.0 h, which is close to that observed in chow-fed animals. In contrast, farnesol treatment does not affect the turnover rate of reductase protein in rats fed a normal chow diet. These results suggest that potent inhibitors of HMG-CoA reductase may unmask transcriptional regulation by dietary cholesterol and accelerated degradation of the reductase by the putative nonsterol regulator farnesol. PMID- 9210654 TI - Expression, purification, and enzymatic properties of recombinant human cytochrome P450c27 (CYP27). AB - A large number of microsomal P450s have been expressed in Escherichia coli in quantities sufficient for structure/function analysis. However, only one mitochondrial P450 has been successfully overexpressed, that being cholesterol side chain cleavage cytochrome P450 (P450scc). We report here overexpression, purification, and characterization of a second mitochondrial P450, human sterol C 27 hydroxylase (P450c27). The conditions used for expression are very similar to those applied for P450scc, although a quite different purification protocol was necessary to achieve highly purified P450c27. The catalytic properties of purified recombinant human P450c27 resemble those of purified, endogenous rat and rabbit P450c27, regarding both specificity and turnover numbers. Like endogenous P450c27 from rat and rabbit liver, human recombinant P450c27 is only functional in the presence of adrenodoxin and adrenodoxin reductase and shows no activity in the presence of the microsomal P450 reductase. We conclude that P450c27 is most likely not the 1alpha-hydroxylase of 25-hydroxyvitamin D3, contrary to a previous suggestion (Axen, E., Postlind, H., Sjoberg, H., and Wikvall, K. (1994) Proc. Natl. Acad. Sci. USA 91, 10014-10018) because this activity of P450c27 (28 pmol/min/nmol P450) seems far too low to be physiologically relevant. This activity is 10(3) times lower than the 27-hydroxylase activity toward 5beta cholestane-3alpha,7alpha,12alpha-triol and 40 times lower than the 27 hydroxylation of cholesterol by this enzyme. The development of this expression system and purification procedure creates the potential for structure/function analysis of P450c27, including possible crystallization of this important enzyme. PMID- 9210655 TI - Reactivity of nitrogen monoxide species with NADH: implications for nitric oxide dependent posttranslational protein modification. AB - Nitric oxide (NO.) and NO. donors incite NAD- [i.e., mono(ADP-ribosylation)] and NADH-dependent posttranslational protein modifications by an as yet unknown mechanism. A route of pyridine nucleotide-dependent, NO.-stimulated protein modification has recently been hypothesized [S. Dimmeler, and B. Brune, (1992) Eur. J. Biochem. 210, 305-310; J. S. Stamler (1994) Cell 78, 931-936]. An essential feature of this proposed mechanism is NADH nitrosation, for a nitroso NADH adduct is considered to be a key reactant in the generation of pyridine nucleotide-modified protein. To evaluate at the molecular level the ability of NADH to act as a nitrosation substrate, the potential effects of NO., the nitrosothiols S-nitrosoglutathione and S-nitrosocysteine, the nitrosating agent tert-butyl-nitrite, and the NO. metabolite peroxynitrite on the molecular and functional (i.e., hydride-transfer) properties of NADH have been directly assessed at physiological pH. Exposure of NADH to NO. or nitrosothiol altered neither the hydride-transfer capability of the pyridine nucleotide nor its ultraviolet spectrum in ways suggestive of NADH nitrosation. As determined by NMR spectroscopy, NADH was refractory to the well-recognized nitrosating agent tert butyl nitrite. Consequently, it appears that NADH is an unfavorable substrate for nitrosation under physiological conditions. These data are inconsistent with the proposal that NO. or a NO.-derived nitrosating agent interacts with NADH to generate the nitroso-NADH hypothesized to be essential to NO.-stimulated, pyridine nucleotide-dependent protein modification. Peroxynitrite, a possible source of nitrosating compounds, readily oxidized NADH to NAD, but demonstrated no potential to form a nitroso-NADH adduct. The facility with which NADH is oxidized to NAD has implications for peroxynitrite-mediated tissue damage. PMID- 9210656 TI - Modulation of platelet responses by 2-[3-(bromo-2-oxopropylthio)]adenosine-5' diphosphate involves its binding to as well as covalent modification of an ADP receptor, aggregin. AB - The 2-substituted ADP derivatives are known to activate human blood platelets with varying degrees of potency. For example, 2-(4-bromo-2,3 dioxobutylthio)adenosine-5'-diphosphate [2-BDB-TADP], an ADP-affinity analog, was previously shown by us to be 50% as potent as ADP in inducing human blood platelet responses [Puri, R. N., Colman, R. F., and Colman, R. W. (1996) Eur. J. Biochem. 236, 862-870]. 2-Methylthio-ADP (2-MeS-ADP) has been known to be a far more potent agonist than ADP. However, the molecular basis for defining the rank order of potency of the 2-substituted ADP derivatives as agonists of platelet responses have been incompletely understood. We now report that 2-BOP-TADP (a one carbon atom lower homolog of 2-BDB-TADP) at equimolar concentration is as potent as ADP in inducing platelet responses. Prolonged incubation of platelets with 2 BOP-TADP abolished its ability to elicit cellular responses. An autoradiogram of the gel obtained by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of solubilized platelets labeled by incubating the platelets with 2-BOP-TADP for 1 h followed by reduction by NaB[3H]4 showed the presence of a single covalently radiolabeled protein band at 100 kDa. Preincubation of platelets with either ADP or ATP reduced the intensity of the band corresponding to the 100-kDa protein radiolabeled by 2-BOP-TADP and NaB[3H]4. The results show that (i) 2-BOP-TADP modulates ADP-induced platelet responses by interacting with aggregin and (ii) 2 BOP-TADP was twice as potent as 2-BDB-TADP, and (iii) the chain length of the substituent in a homologous series has an important bearing on the potency of a 2 substituted ADP analog. PMID- 9210657 TI - Transient kinetics of polyamine oxidase from Zea mays L. PMID- 9210659 TI - Synergistic anti-influenza virus A (H1N1) activities of PM-523 (polyoxometalate) and ribavirin in vitro and in vivo. AB - A Kegin-type polyoxometalate, PM-523, in combination with ribavirin, was tested for its therapeutic effectiveness against influenza virus (FluV) A (H1N1) infection in tissue culture and in mice. PM-523 [(PriNH3)6H [PTi2W10O38(O2)2] x H2O, where Pri is isopropanol] and ribavirin individually inhibited FluV A induced cytopathic effects in Madin-Darby canine kidney (MDCK) cells at median effective concentrations (EC50s) of 30 and 34 microM, respectively, and at 70% effective concentrations (EC70s) of 48 and 72 microM, respectively. On the other hand, a combination of PM-523 and ribavirin at a ratio of 1:16 exhibited lower EC50s and EC70s than each compound used singly, and combination indices were less than 1. A wide range of combinations of PM-523 and ribavirin at ratios of from 1:128 to 1:1 exhibited additive or synergistic anti-FluV effects in MDCK cells. When these compounds were tested for their anti-FluV A activities in vivo by aerosol exposure of mice which had been infected with a lethal dose of FluV A by an intranasal route, a 1:16 combination of PM-523 and ribavirin was found to have a significantly better therapeutic effect than a single dose of either compound used singly with respect to both the survival rate of the mice and the virus titer in the lungs of the infected mice. PM-523 was effective for the treatment of experimental FluV infection, and in combination with ribavirin, PM-523 exhibited enhanced anti-FluV effects in vitro and in vivo compared with the effect of PM-523 alone. PMID- 9210658 TI - Assessment of the pharmacodynamic properties of antimalarial drugs in vivo. PMID- 9210660 TI - Inhibitors of delta24(25) sterol methyltransferase block sterol synthesis and cell proliferation in Pneumocystis carinii. AB - Detailed analysis of the endogenous sterol content of purified Pneumocystis carinii preparations by gas-liquid chromatography coupled to mass spectrometry suggested that this parasite can both synthesize de novo steroid skeletons (to produce delta7 sterols) and take them from the infected host (leading to delta5 sterols). In both cases the final products are 24-alkyl sterols, resulting from the action of delta24(25) and delta24(24') sterol methyltransferases, enzymes not present in vertebrates. To investigate the physiological significance of these sterols, cultures of P. carinii in embryonic lung cells were exposed to 22,26 azasterol (20-piperidin-2-yl-5alpha-pregnan-3beta-20(R)-diol), a compound previously shown to inhibit both enzymes and to halt cell proliferation in fungi and protozoa. This compound produced a dose-dependent reduction in the parasite proliferation, with a 50% inhibitory concentration of 0.3 microM and 80% reduction of growth after 96 h at 10 microM. Correspondingly, parasites treated with the azasterol at 10 microM for 48 h accumulated 24-desalkyl sterols such as zymosterol (cholesta-8,24-dien-3beta-ol) and cholesta-8,14,24-trien-3beta-ol to ca. 40% of the total mass of endogenous sterols. This is the first report on the antiproliferative effects of a sterol biosynthesis inhibitor on P. carinii and indicate that sterol methyltransferase inhibitors could be the basis of a novel and specific chemotherapeutic approach to the treatment of P. carinii infections. PMID- 9210661 TI - Antimicrobial activities of squalamine mimics. AB - We investigated the antimicrobial properties of compounds with structural features that were designed to mimic those of squalamine, an antibiotic isolated from the stomach of the dogfish shark. The mimics, like squalamine, are sterol polyamine conjugates. Unlike squalamine, the mimics were simple to prepare, at high yield, from readily available starting materials. Several squalamine mimics showed activity against gram-negative rods, gram-positive cocci including methicillin-resistant Staphylococcus aureus, vancomycin-resistant Enterococcus faecium, and fungi. Some had little or no hemolytic activity. The hydrophobicity of the sterol backbone and the length and the cationic charge of the side chains appeared to be critical determinants of activity. One of the squalamine mimics, SM-7, was bactericidal against Escherichia coli, Pseudomonas aeruginosa, and S. aureus; its activity was decreased by divalent or monovalent cations and by bovine serum albumin. Subinhibitory concentrations of SM-7 markedly enhanced the antimicrobial activity of rifampin against gram-negative rods. These results suggest that the compounds may disrupt an outer membrane of gram-negative rods. Squalamine mimics are a new class of broad-spectrum antimicrobial agents. The antagonism of their activity by serum and albumin and their hemolytic properties may limit their use as systemic agents. The squalamine mimics, because of their potencies, broad spectra of antimicrobial activity, and potential for systemic toxicity, appear to be good candidates for development as topical antimicrobial agents. PMID- 9210662 TI - Differential induction of pro- and anti-inflammatory cytokines in whole blood by bacteria: effects of antibiotic treatment. AB - The in vitro production of interleukin-1beta (IL-1beta), IL-6, and the IL-1 receptor antagonist (IL-1ra) in whole blood upon stimulation with different bacterial strains was measured to study the possible relationship between disease severity and the cytokine-inducing capacities of these strains. Escherichia coli, Neisseria meningitidis, Neisseria gonorrhoeae, Bacteroides fragilis, Capnocytophaga canimorsus, Staphylococcus aureus, Enterococcus faecalis, Streptococcus pneumoniae, and Streptococcus pyogenes induced the cytokines IL 1beta, IL-6, and IL-1ra. Gram-negative bacteria induced significantly higher levels of proinflammatory cytokine production than gram-positive bacteria. These differences were less pronounced for the anti-inflammatory cytokine IL-1ra. In addition, blood was stimulated with E. coli killed by different antibiotics to study the effect of the antibiotics on the cytokine-inducing capacity of the bacterial culture. E. coli treated with cefuroxime and gentamicin induced higher levels of IL-1beta and IL-6 production but levels of IL-1ra production similar to that of heat-killed E. coli. In contrast, ciprofloxacin- and imipenem-cilastatin mediated killing showed a decreased or similar level of induction of cytokine production as compared to that by heat-killed E. coli; polymyxin B decreased the level of production of the cytokines. PMID- 9210663 TI - Identification of BMS-200475 as a potent and selective inhibitor of hepatitis B virus. AB - BMS-200475 is a novel carbocyclic 2'-deoxyguanosine analog found to possess potent and selective anti-hepatitis B virus (anti-HBV) activity. BMS-200475 is distinguished from guanosine by replacement of the natural furanose oxygen on the sugar moiety with an exo carbon-carbon double bond. In the HepG2 stably transfected cell line 2.2.15, BMS-200475 had a 50% effective concentration (EC50) of 3.75 nM against HBV, as determined by analysis of secreted HBV DNA. Structurally related compounds with adenine, iodouracil, or thymine base substitutions were significantly less potent or were inactive. Direct comparison of the antiviral activities of BMS-200475 with those of a variety of other nucleoside analogs, including lamivudine (EC50 = 116.26 nM), demonstrated the clearly superior in vitro potency of BMS-200475 in 2.2.15 cells. Intracellular HBV replicative intermediates were uniformly reduced when cells were treated with BMS-200475, but rebounded after treatment was terminated. The concentration of BMS-200475 causing 50% cytotoxicity in 2.2.15 cell cultures was 30 microM, approximately 8,000-fold greater than the concentration required to inhibit HBV replication in the same cell line. Treatment with BMS-200475 resulted in no apparent inhibitory effects on mitochondrial DNA content. PMID- 9210664 TI - Potentiation of an antimalarial oxidant drug. AB - In a previous report we described the synergistic antimalarial interaction between two structurally similar compounds, rufigallol and exifone. To explain this phenomenon, we proposed that exifone is transformed inside the parasitized erythrocyte into a xanthone with potent antimalarial properties. We speculated that the transformation process was induced by the prooxidant activity of rufigallol. On the basis of this model we hypothesized that exifone would act synergistically with other oxidant drugs. In the present study we have found a similar synergistic interaction between exifone and ascorbic acid (vitamin C) against both chloroquine-susceptible and multidrug-resistant strains of Plasmodium falciparum. The prooxidant activity of ascorbic acid against Plasmodium-infected erythrocytes is believed to result from an intraerythrocytic Fenton reaction occurring in the acidic food vacuole of the parasite. The hydroxyl radicals produced during this process are believed to attack exifone, which undergoes cyclodehydration to become 2,3,4,5,6-pentahydroxyxanthone (X5). Evidence presented to support this "xanthone hypothesis" includes the demonstration that the exifone ==> X5 transformation occurs readily in vitro under mildly acidic conditions in the presence of iron, ascorbic acid, and oxygen. PMID- 9210665 TI - Efficacy of D0870 treatment of experimental Candida vaginitis. AB - In this study, oral administration of the triazole D0870 was compared to oral administration of fluconazole in the treatment of experimental vaginal candidiasis. With an estrogen-dependent murine model of Candida albicans vaginal infection, the effects of D0870 on several isolates, including fluconazole susceptible and -resistant isolates, were tested. D0870, at doses of 0.5 and 2.5 mg/kg of body weight given once over the course of a 10-day infection, was effective in eradicating vaginitis caused by fluconazole-susceptible laboratory and clinical isolates, respectively. In contrast, a stricter treatment regimen (every 24 to 48 h) with 10 and 25 mg of fluconazole per kg was required to achieve similar reductions in vaginal fungal titers induced by the same isolates. Whereas fluconazole was consistently ineffective in infections induced by fluconazole-resistant isolates, as predicted by in vitro susceptibility tests, D0870 was effective, although a daily regimen of 25 mg/kg was required. Additional studies showed that despite the in vitro activity of D0870 against two clinical Candida glabrata isolates, neither D0870 nor fluconazole was effective at daily doses as high as 100 and 125 mg/kg, respectively. Taken together, although D0870 failed to show efficacy against experimental C. glabrata vaginitis, D0870 was superior to fluconazole in the treatment of experimental C. albicans vaginitis caused by isolates that were either susceptible or resistant to fluconazole. PMID- 9210666 TI - Sequence analysis and enzyme kinetics of the L2 serine beta-lactamase from Stenotrophomonas maltophilia. AB - The L2 serine active-site beta-lactamase from Stenotrophomonas maltophilia has been classified as a clavulanic acid-sensitive cephalosporinase. The gene encoding this enzyme from S. maltophilia 1275 IID has been cloned on a 3.3-kb fragment into pK18 under the control of a Ptac promoter to generate recombinant plasmid pUB5840; when expressed in Escherichia coli, this gene confers resistance to cephalosporins and penicillins. Sequence analysis has revealed an open reading frame (ORF) of 909 bp with a GC content of 71.6%, comparable to that of the L1 metallo-beta-lactamase gene (68.4%) from the same bacterium. The ORF encodes an unmodified protein of 303 amino acids with a predicted molecular mass of 31.5 kDa, accommodating a putative leader peptide of 27 amino acids. Comparison of the amino acid sequence with those of other beta-lactamases showed it to be most closely related (54% identity) to the BLA-A beta-lactamase from Yersinia enterocolitica. Sequence identity is most obvious near the STXK active-site motif and the SDN loop motif common to all serine active-site penicillinases. Sequences outside the conserved regions display low homology with comparable regions of other class A penicillinases. Kinetics of the enzyme from the cloned gene demonstrated an increase in activity with cefotaxime but markedly less activity with imipenem than previously reported. Hence, the S. maltophilia L2 beta lactamase is an inducible Ambler class A beta-lactamase which would account for the sensitivity to clavulanic acid. PMID- 9210667 TI - Stereoselective interaction of the azole antifungal agent SCH39304 with the cytochrome P-450 monooxygenase system isolated from Cryptococcus neoformans. AB - We investigated the stereoselective inhibition of growth and ergosterol biosynthesis by SCH39304 in the pathogenic fungus Cryptococcus neoformans obtained from four AIDS patients who failed fluconazole therapy and compared the results to those obtained with a wild-type strain. For all strains, the MICs of the RR isomer were approximately half those of the racemate, with the SS enantiomer showing no inhibitory activity. The 50% inhibitory concentrations for in vitro ergosterol biosynthesis correlated with the MIC data, indicating stereoselective inhibition of their target P-450 enzyme, sterol 14alpha demethylase, as the cause of this difference. The RR enantiomer produced classical type II spectra on addition to microsomal extracts of the strains, whereas the SS enantiomer showed an absence of binding. Stereo- and regio specific localization of N-1 substituent groups of SCH39304 within the active site of the enzyme determined the unique discrimination between its two enantiomers, and the inability to bind to sterol 14alpha-demethylase is also true of other P-450 enzymes contained in the microsomal fraction. As previously observed for other antifungal azoles, isolates obtained following failure of fluconazole therapy showed resistance to SCH39304 and its RR enantiomer. This resistance could be associated with an alteration in the sensitivity of ergosterol biosynthesis in vitro. These alterations did not cause any changes allowing the SS enantiomer to bind to the P-450 mediating sterol 14alpha demethylation. PMID- 9210668 TI - Time-restricted feeding schedules modify temporal variation of gentamicin experimental nephrotoxicity. AB - The effect of timing of gentamicin dosing relative to food access periods was evaluated in experimental animals. Female Sprague-Dawley rats were treated for 4 and 10 days with gentamicin (40 mg/kg of body weight/day) intraperitoneally at either 0700, 1300, 1900, or 0100 h according to three food presentation schedules: food was available from 0800 to 1600 h in the first group, from 1600 to 0000 h in the second group, and from 0000 to 0800 h in the last group. Animals were thus subjected to a restricted feeding period. Results indicate that time restricted feeding schedules displace the peak and the trough of gentamicin induced renal toxicity, as evaluated by changes in the inhibition of sphingomyelinase activity, cellular regeneration (incorporation of [3H]thymidine into DNA of renal cortex), and blood urea nitrogen and serum creatinine levels, as well as histopathological lesions observed after 10 days of treatment. In fact, the toxicity was minimal when gentamicin was injected during the feeding period, while the maximal toxicity was found when gentamicin was administered during the fasting period. It is concluded that the feeding period can modulate aminoglycoside nephrotoxicity. The time of dosing of gentamicin relative to the time of feeding seems to be a more important modulator of gentamicin nephrotoxicity than the light-dark cycle. PMID- 9210669 TI - Comparative activity of trovafloxacin, alone and in combination with other agents, against gram-negative nonfermentative rods. AB - In the first part of this study, agar dilution MICs were used to test the activities of trovafloxacin, ciprofloxacin, ofloxacin, levofloxacin, sparfloxacin, clinafloxacin, ceftazidime, and imipenem against 458 gram-negative nonfermenters. The overall respective MICs at which 50% of isolates are inhibited (MIC50s) and MIC90s were as follows: trovafloxacin, 1.0 and 16.0 microg/ml; ciprofloxacin, 2.0 and 16.0 microg/ml; ofloxacin, 2.0 and 32.0 microg/ml; levofloxacin, 1.0 and 16.0 microg/ml; sparfloxacin, 1.0 and 16.0 microg/ml; clinafloxacin, 0.5 and 4.0 microg/ml; ceftazidime, 8.0 and 128.0 microg/ml; imipenem, 2.0 and 256.0 microg/ml. Clinafloxacin was the most active of all the quinolones tested. The MIC90s of trovafloxacin were < or = 4.0 microg/ml for Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Flavobacterium odoratum, and Chryseobacterium meningosepticum; trovafloxacin MIC90s were < or = 2.0 microg/ml for Moraxella spp., Pseudomonas stutzeri, and Chryseobacterium indologenes-C. gleum. Of the other quinolones tested, the MICs of sparfloxacin and levofloxacin were lower than those of ciprofloxacin and ofloxacin. High ceftazidime MICs (> or = 32.0 microg/ml) were observed for all nonfermentative species tested. Although for the majority of strains tested imipenem MICs were < or = 8.0 microg/ml, high imipenem MICs were observed for many species, especially S. maltophilia, Burkholderia cepacia, F. odoratum, and Chryseobacterium meningosepticum. For Alcaligenes xylosoxidans strains, the MICs of all compounds were generally a few dilutions lower than those for Alcaligenes faecalis-A. odorans. Time-kill studies with five strains revealed that trovafloxacin and all quinolones yielded more rapid time-kill kinetics than ceftazidime and imipenem. Synergy testing by checkerboard titrations of 286 strains with trovafloxacin combined with ceftazidime, amikacin, and imipenem revealed fractional inhibitory concentration (FIC) indices in the range indicating synergism (< or = 0.5) for 81, 41, and 40 strains, respectively, and FIC indices indicating additivity or indifference (> 0.5 to 4.0) for 205, 245, and 246 strains, respectively. No FIC indices indicating antagonism (> 4.0) were observed. Synergy between trovafloxacin and ceftazidime was found for 32 of 36 S. maltophilia strains. Time kill studies with 20 strains showed that for most strains for which FIC indices were in the range indicating additivity or indifference, FIC indices indicated synergy by the time-kill method. Synergy was particularly noticeable for S. maltophilia strains with combinations of ceftazidime and trovafloxacin. PMID- 9210670 TI - Increased mRNA levels of ERG16, CDR, and MDR1 correlate with increases in azole resistance in Candida albicans isolates from a patient infected with human immunodeficiency virus. AB - Resistance to antifungal drugs, specifically azoles such as fluconazole, in the opportunistic yeast Candida albicans has become an increasing problem in human immunodeficiency virus (HIV)-infected individuals. The molecular mechanisms responsible for this resistance have only recently become apparent and can include alterations in the target enzyme of the azole drugs (lanosterol 14alpha demethylase [14DM]), or in various efflux pumps from both the ABC transporter and major facilitator gene families. To determine which of these possible mechanisms was associated with the development of drug resistance in a particular case, mRNA levels have been studied in a series of 17 clinical isolates taken from a single HIV-infected patient over 2 years, during which time the levels of fluconazole resistance of the strain increased over 200-fold. Using Northern blot analysis of steady-state levels of total RNA from these isolates, we observed increased mRNA levels of ERG16 (the 14DM-encoding gene), CDR1 (an ABC transporter), and MDR1 (a major facilitator) in this series. The timing of the increase in mRNA levels of each of these genes correlated with increases in fluconazole resistance of the isolates. Increased mRNA levels were not observed for three other ABC transporters, two other genes in the ergosterol biosynthetic pathway, or the NADPH-cytochrome P-450 oxidoreductase gene that transfers electrons from NADPH to 14DM. Increases in mRNA levels of ERG16 and CDR1 correlated with increased cross resistance to ketoconazole and itraconazole but not to amphotericin B. A compilation of the genetic alterations identified in this series suggests that resistance develops gradually and is the sum of several different changes, all of which contribute to the final resistant phenotype. PMID- 9210671 TI - The presence of an R467K amino acid substitution and loss of allelic variation correlate with an azole-resistant lanosterol 14alpha demethylase in Candida albicans. AB - Azole resistance in the pathogenic yeast Candida albicans is an emerging problem in the human immunodeficiency virus (HIV)-infected population. The target enzyme of the azole drugs is lanosterol 14alpha demethylase (Erg16p), a cytochrome P-450 enzyme in the biosynthetic pathway of ergosterol. Biochemical analysis demonstrates that Erg16p became less susceptible to fluconazole in isolate 13 in a series of isolates from an HIV-infected patient. PCR-single-strand conformation polymorphism (PCR-SSCP) analysis was used to scan for genomic alterations of ERG16 in the isolates that would cause this change in the enzyme in isolate 13. Alterations near the 3' end of the gene that were identified by PCR-SSCP were confirmed by DNA sequencing. A single amino acid substitution (R467K) that occurred in isolate 13 was identified in both alleles of ERG16. Allelic differences within the ERG16 gene, in the ERG16 promoter, and in the downstream THR1 gene were eliminated in isolate 13. The loss of allelic variation in this region of the genome is most likely the result of mitotic recombination or gene conversion. The R467K mutation and loss of allelic variation that occur in isolate 13 are likely responsible for the azole-resistant enzyme activity seen in this and subsequent isolates. The description of R467K represents the first point mutation to be identified within ERG16 of a clinical isolate of C. albicans that alters the fluconazole sensitivity of the enzyme. PMID- 9210672 TI - Metabolism of a 5-nitroimidazole in susceptible and resistant isogenic strains of Bacteroides fragilis. AB - We investigated the metabolism of dimetridazole (1,2-dimethyl-5-nitroimidazole) (DMZ) by the resting cell method in a susceptible strain of Bacteroides fragilis and in the same strain containing the nimA gene, which conferred resistance to 5 nitroimidazole drugs. In both cases, under strict anaerobic conditions DMZ was metabolized without major ring cleavage or nitrate formation. However, one of two distinct metabolic pathways is involved, depending on the susceptibility of the strain. In the susceptible strain, the classical reduction pathway of nitroaromatic compounds is followed at least as far as the nitroso-radical anion, with further formation of the azo-dimer: 5,5'-azobis-(1,2-dimethylimidazole). In the resistant strain, DMZ is reduced to the amine derivative, namely, 5-amino-1,2 dimethylimidazole, preventing the formation of the toxic form of the drug. The specificity of the six-electron reduction of the nitro group, which is restricted to 4- and 5-nitroimidazole, suggests an enzymatic reaction. We thus conclude that nimA and related genes may encode a 5-nitroimidazole reductase. PMID- 9210673 TI - In vivo and in vitro antiplasmodial activities of some plants traditionally used in Guatemala against malaria. AB - We present an evaluation of the antiplasmodial and cytotoxic effects of four plants commonly used in Guatemalan folk medicine against malaria. Methanol extracts of Simarouba glauca D. C., Sansevieria guineensis Willd, Croton guatemalensis Lotsy, and Neurolaena lobata (L.)R.Br. significantly reduced parasitemias in Plasmodium berghei-infected mice. Dichloromethane fractions were screened for their cytotoxicities on Artemia salina (brine shrimp) larvae, and 50% inhibitory concentrations were determined for Plasmodium falciparum in in vitro cultures. Both chloroquine-susceptible and -resistant strains of P. falciparum were significantly inhibited by these extracts. Of all dichloromethane extracts, only the S. glauca cortex extract was considered to be toxic to nauplii of A. salina in the brine shrimp test. PMID- 9210674 TI - Efficacy of SCH-56592 in a temporarily neutropenic murine model of invasive aspergillosis with an itraconazole-susceptible and an itraconazole-resistant isolate of Aspergillus fumigatus. AB - SCH-56592 (SCH) is a novel triazole antifungal agent with excellent in vitro activity against Aspergillus. We compared three doses (5, 10, and 25 mg/kg of body weight) of SCH with itraconazole (ITZ; 25 mg/kg) and amphotericin B (AB; 5 mg/kg) in a temporarily neutropenic murine model of disseminated aspergillosis (lungs and kidneys) against one ITZ-susceptible (AF71) and one ITZ-resistant (AF90) isolate of Aspergillus fumigatus. Treatment started 24 h after infection and lasted for 10 days. Dosing regimens for SCH were once daily for 10 days, those for ITZ were three times daily for 2 days and then twice daily for 3 to 10 days, and those for AB were once daily on days 1, 2, 4, and 7. Both isolates killed 90% of control mice. Kidneys and lungs from survivors were cultured on day 11. Against AF71, all three doses of SCH and ITZ yielded a 90 to 100% survival rate and AB yielded 40% survival (P < or = 0.01 to 0.0001 for all treatment groups compared with the controls). All three doses of SCH were superior to AB in cultures of lung and kidney tissue samples (P < or = 0.01 to 0.0002) and SCH at 25 mg/kg was superior to ITZ in cultures of kidneys (P = 0.01). Against AF90, the highest dose of SCH (25 mg/kg) resulted in a 100% survival rate, compared with 60 and 20% survival rates for the groups treated with SCH at 10 and 5 mg/kg, respectively. Treatment with ITZ yielded no survivors. AB therapy achieved a 50% survival rate. SCH at 25 mg/kg (P < 0.001), SCH at 10 mg/kg (P < or = 0.005), and AB (P < 0.05) were superior to ITZ in cultures of lungs and kidneys. There was a correlation between the MICs of SCH and quantitative organ culture results and between the minimum fungicidal concentration of AB with quantitative organ culture results. SCH appears to be a highly effective anti-Aspergillus compound in this model. There appears to be a degree of cross-resistance between itraconazole and SCH. PMID- 9210675 TI - Oral bioavailability and pharmacokinetics of ciprofloxacin in patients with AIDS. AB - Few reports on the effects of AIDS on the absorption of orally (p.o.) administered agents exist. To help fill this informational gap, we administered ciprofloxacin to 12 patients with AIDS by two dosing regimens (400 mg given intravenously [i.v.] and 500 mg given p.o. every 12 h) in a randomized, crossover fashion. Pharmacokinetic parameters were determined by noncompartmental methods. Mean values (+/- standard deviations [SD]) for p.o. ciprofloxacin were as follows: peak concentration of drug in serum (Cmax), 2.94 +/- 0.51 microg/ml; time to Cmax, 1.38 +/- 0.43 h; area under the concentration-time curve from 0 to 12 h (AUC(0-12)), 12.13 +/- 3.21 microg x h/ml; and half-life (t(1/2)), 3.86 +/- 0.48 h. Mean values (+/- SD) for i.v. ciprofloxacin were as follows: Cmax, 3.61 +/- 0.82 microg/ml; time to Cmax, 1.0 h; AUC(0-12), 11.92 +/- 2.92 microg x h/ml; and t(1/2), 3.98 +/- 0.94 h. The mean percent absolute bioavailability for ciprofloxacin was calculated to be 82% +/- 13%, similar to the value for healthy volunteers. We conclude that ciprofloxacin when administered p.o. to patients with AIDS is well absorbed, as evidenced by excellent bioavailability and is not affected by gastrointestinal changes in the absence of infectious gastroenteritis and severe diarrhea. PMID- 9210676 TI - In vitro reduction of endotoxin concentrations with the 5S fragment of immunoglobulin G. AB - Endotoxin has long been implicated as an inducer for the development and progression of gram-negative sepsis. Accordingly, antiendotoxin therapy has been considered one of the major targets for the treatment of sepsis. To investigate the influence of a human immunoglobulin G (IgG) derivative, the 5S fragment of IgG (5S-IgG; Gamma-Venin, Centeon Pharma GmbH, Frankfurt-Niederrad, Germany), on endotoxin release during bacterial proliferation and under antibiotic bactericidal action, time-kill studies were performed by using Escherichia coli ATCC 25922 starting inocula of 10(3), 10(5), and 10(7) CFU/ml with cefotaxime (120 microg/ml) alone and in combination with 5S-IgG (2,100 microg/ml). Samples were collected for bacterial colony count and endotoxin concentration determinations; the area under the free endotoxin concentration curve (AUFEC) was calculated by using the trapezoidal rule. Colony counts showed that cefotaxime had a rapid bactericidal effect because it achieved greater than a 4-log decrease in the numbers of E. coli CFU per milliliter over the first 2 h; the addition of 5S-IgG did not appear to alter the kinetics of killing. Comparison of the AUFEC revealed that the addition of 5S-IgG resulted in a mean reduction of 50, 66, and 27% in the free endotoxin concentration at starting inocula of 10(3), 10(5), and 10(7) CFU/ml, respectively. Moreover, experiments were conducted with a starting inoculum of 10(5) CFU/ml and various amounts of 5S-IgG (2 to 20 mg/ml) to further investigate the dose-effect relation of 5S-IgG on endotoxin release. Decreased AUFECs were observed with increasing concentrations of 5S-IgG, suggesting the dose-dependent antiendotoxin activity of 5S-IgG. Further study is required to investigate the mechanism(s) responsible for this observation, the biological significance of this antiendotoxin activity, and the potential utility of 5S-IgG as an adjuvant therapy in the treatment of gram-negative sepsis. PMID- 9210677 TI - Comparative effectiveness and safety of cefdinir and amoxicillin-clavulanate in treatment of acute community-acquired bacterial sinusitis. Cefdinir Sinusitis Study Group. AB - Cefdinir is an extended-spectrum oral cephalosporin that is active against pathogens commonly seen in acute community-acquired bacterial sinusitis (ACABS), including Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. Two randomized, investigator-blind, multicenter trials (one in the United States and one in Europe) compared two dosage regimens of cefdinir (600 mg once a day for 10 days and 300 mg twice a day for 10 days) to amoxicillin clavulanate (A-C) (500 mg three times a day for 10 days) for adult and adolescent patients with ACABS. Twelve hundred twenty-nine patients entered the U.S. study, 698 with antral puncture; 569 patients entered the European study, all with antral puncture. Clinical response (cure or improvement) was determined 7 to 14 days and 3 to 5 weeks posttherapy. Microbiologic eradication rates were determined 10 to 30 days posttherapy in a subset of patients who underwent pre- and posttherapy sinus aspirate culture. Rates of adverse events and treatment discontinuations due to adverse events were examined. Cefdinir, given once or twice daily, was as effective clinically (approximately 90% cure rate) as amoxicillin-clavulanate given three times daily in the treatment of ACABS. Microbiologic eradication rates were also similar in the three groups. The major side effect was mild diarrhea, occurring in approximately 20% of each group. Cefdinir caused fewer adverse events requiring treatment discontinuation. PMID- 9210679 TI - Effects of 2',3'-dideoxyinosine on Toxoplasma gondii cysts in mice. AB - The activity against Toxoplasma gondii of 2',3' dideoxyinosine (ddI), an anti human immunodeficiency virus drug, was examined in an in vitro and in vivo study. Cell cultures infected with a strain known to cause chronic infections were used to show the dose-dependent effect of this drug compared with spiramycin and sulfadiazine. When a dose of 4 microg/ml was used, no infected THP-1 cells or parasites were found after 60 h of incubation. An electron-microscopic study confirmed that after 12 h at 1 microg/ml, the few parasites observed were severely altered. The treatment of chronically infected mice 3 months postinfection showed that a 30-day treatment with 2 mg of ddI/ml induced a significant reduction in the number of T. gondii cysts in the cerebral tissue. These cysts were not viable, as confirmed by immunofluorescence and reinfection experiments. These experiments suggest a possible role for ddI in the treatment of toxoplasmosis, and this possibility deserves further investigation. PMID- 9210678 TI - Discovery of cyanovirin-N, a novel human immunodeficiency virus-inactivating protein that binds viral surface envelope glycoprotein gp120: potential applications to microbicide development. AB - We have isolated and sequenced a novel 11-kDa virucidal protein, named cyanovirin N (CV-N), from cultures of the cyanobacterium (blue-green alga) Nostoc ellipsosporum. We also have produced CV-N recombinantly by expression of a corresponding DNA sequence in Escherichia coli. Low nanomolar concentrations of either natural or recombinant CV-N irreversibly inactivate diverse laboratory strains and primary isolates of human immunodeficiency virus (HIV) type 1 as well as strains of HIV type 2 and simian immunodeficiency virus. In addition, CV-N aborts cell-to-cell fusion and transmission of HIV-1 infection. Continuous, 2-day exposures of uninfected CEM-SS cells or peripheral blood lymphocytes to high concentrations (e.g., 9,000 nM) of CV-N were not lethal to these representative host cell types. The antiviral activity of CV-N is due, at least in part, to unique, high-affinity interactions of CV-N with the viral surface envelope glycoprotein gp120. The biological activity of CV-N is highly resistant to physicochemical denaturation, further enhancing its potential as an anti-HIV microbicide. PMID- 9210680 TI - Evaluation of a flow cytofluorometric method for rapid determination of amphotericin B susceptibility of yeast isolates. AB - A rapid-flow cytofluorometric susceptibility test for in vitro amphotericin B testing of yeasts was evaluated and compared to the National Committee for Clinical Laboratory Standards (NCCLS) M27-T reference broth macrodilution method. The flow cytofluorometric method is based on the detection of decreased green fluorescence intensity of cells stained with DiOC5(3), a membrane potential sensitive cationic dye, after drug treatment. Testing was performed on 134 clinical isolates (Candida spp. and Torulopsis glabrata). From the dose-response curve obtained for each isolate, three endpoints were calculated by computer analysis (the concentrations at which the fluorescence intensity was reduced by 50, 80, and 90%, i.e., 50% inhibitory concentration [IC50], IC80, and IC90, respectively). A regression analysis correlating these endpoints with the M27-T MICs showed that the best agreement was obtained with IC80. The flow cytofluorometric method showed good reproducibility with control strains. These initial results suggest that the flow cytofluorometric method is a valid alternative to the NCCLS reference method. PMID- 9210682 TI - Differential intracellular efficacies of ciprofloxacin and cefixime against Neisseria gonorrhoeae in human fallopian tube organ culture. AB - This study compared the abilities of ciprofloxacin and cefixime to kill intracellular Neisseria gonorrhoeae in a human fallopian tube organ culture assay. When invasion was inhibited by cytochalasin D, 0.996% of the tissue associated gonococci survived ciprofloxacin exposure compared to 1.70% of gonococci exposed to cefixime (95% confidence interval for the ratio of the means, 0.267 to 1.30), indicating that the two antibiotics did not significantly differ in the ability to kill extracellular attached organisms. In the absence of cytochalasin D, 1.63% survived ciprofloxacin exposure while 9.76% survived cefixime treatment (95% confidence interval for the ratio of the means, 0.067 to 0.418). These results suggest that ciprofloxacin penetrated epithelial cells and killed intracellular gonococci better than did cefixime. Thus, at concentrations achievable in serum, ciprofloxacin was more effective in total gonococcal killing than cefixime in this human fallopian tube organ culture model. PMID- 9210681 TI - Effects of albendazole, fumagillin, and TNP-470 on microsporidial replication in vitro. AB - Presently, the two most commonly used drugs for treating microsporidiosis in persons with AIDS are albendazole and fumagillin. Albendazole is effective for treating disseminated infections due to Encephalitozoon spp. but is variably effective against Enterocytozoon bieneusi infections. Fumagillin is highly effective when used topically to treat ocular infections with Encephalitozoon hellem or Encephalitozoon intestinalis but is too toxic for systemic use. In this study, the fumagillin analog TNP-470 was assayed for antimicrosporidial activity in vitro. The MICs of TNP-470 at which 50% of isolates were killed (MIC50s) were 0.35 +/- 0.21 and 0.38 +/- 0.11 ng/ml for E. intestinalis and Vittaforma corneae, respectively, and were similar to the MIC50s of fumagillin for these organisms, which were 0.515 +/- 0.002 and 0.81 +/- 0.014 ng/ml, respectively. The MIC50 of albendazole for E. intestinalis was 8.0 +/- 4.23 ng/ml, significantly less (P < 0.01) than its MIC50 for V. corneae, which was 55.0 +/- 7.07 ng/ml. TNP-470 inhibited replication of E. intestinalis in RK-13 cells if it was given at the same time as infection or if treatment was initiated 7 days later. In addition, treatment of the infected cultures with TNP-470 at a dose of 10 ng/ml for 2 weeks, followed by discontinuation of the drug treatment, resulted in no significant increase in E. intestinalis shedding during the following 3 weeks in culture. Because TNP-470 acts against both E. intestinalis and V. corneae, and because TNP-470 was found by others to be less toxic in vivo, TNP-470 may be a promising new drug for the treatment of microsporidiosis. PMID- 9210683 TI - In vitro activity of Bay 12-8039, a new 8-methoxyquinolone, compared to the activities of 11 other oral antimicrobial agents against 390 aerobic and anaerobic bacteria isolated from human and animal bite wound skin and soft tissue infections in humans. AB - The in vitro activity of Bay 12-8039, a new oral 8-methoxyquinolone, was compared to the activities of 11 other oral antimicrobial agents (ciprofloxacin, levofloxacin, ofloxacin, sparfloxacin, azithromycin, clarithromycin, amoxicillin clavulanate, penicillin, cefuroxime, cefpodoxime, and doxycycline) against 250 aerobic and 140 anaerobic bacteria recently isolated from animal and human bite wound infections. Bay 12-8039 was active against all aerobic isolates, both gram positive and gram-negative isolates, at < or = 1.0 microg/ml (MICs at which 90% of isolates are inhibited [MIC90s < or = 0.25 microg/ml) and was active against most anaerobes at < or = 0.5 microg/ml; the exceptions were Fusobacterium nucleatum and other Fusobacterium species (MIC90s, > or = 4.0 microg/ml) and one strain of Prevotella loeschii (MICs, 2.0 microg/ml). In comparison, the other quinolones tested had similar in vitro activities against the aerobic strains but were less active against the anaerobes, including peptostreptococci, Porphyromonas species, and Prevotella species. The fusobacteria were relatively resistant to all the antimicrobial agents tested except penicillin G (one penicillinase-producing strain of F. nucleatum was found) and amoxicillin clavulanate. PMID- 9210684 TI - Activity of the triazole SCH 56592 against disseminated murine coccidioidomycosis. AB - SCH 56592 (SCH) is a new triazole antifungal with a broad spectrum of activity. In vitro susceptibility testing against five strains of Coccidioides immitis revealed MICs from 0.39 to 3.13 microg/ml and minimal fungicidal concentrations from 1.56 to 3.13 microg/ml. A murine model of systemic coccidioidomycosis was established in female CD-1 mice. Groups received either no treatment or oral therapy with fluconazole at 10 or 100 mg/kg of body weight; itraconazole at 10 or 100 mg/kg; SCH at 0.5, 2, 10, or 25 mg/kg; or its methylcellulose diluent alone. Therapy began 2 days postinfection and continued once daily for 19 days. Surviving mice were euthanized 49 days postinfection, and infectious burdens were determined by culture. All drugs were superior to no-treatment or diluent treatment controls (P < 0.001) in prolonging survival but were not significantly different from one another. Itraconazole at 100 mg/kg was superior to fluconazole in reduction of CFU in the spleen, liver, and lung (P < 0.01 to 0.001). SCH at 0.5 mg/kg was superior to either fluconazole or itraconazole at 10 mg/kg in reduction of CFU in all three organs (P < 0.05 to 0.001). SCH at 2 mg/kg was not significantly different from itraconazole at 100 mg/kg in all three organs. SCH at 10 and 25 mg/kg was superior to either dose of fluconazole or itraconazole in all three organs (P < 0.05 to 0.001). In terms of reduction of CFU, SCH was > or = 200-fold as potent as fluconazole and > or = 50-fold as potent as itraconazole. There was a clear dose-responsive relationship for SCH in each of the organs. It is noteworthy that SCH effected cures (no detectable C. immitis in any organ) in 1 of 9, 6 of 10, or 9 of 9 surviving mice in animals given 2, 10, or 25 mg/kg, respectively. Neither fluconazole nor itraconazole cured any survivor. SCH has potent, fungicidal activity in vivo against C. immitis. It should be considered for clinical trials in patients with coccidioidomycosis. PMID- 9210685 TI - Absence of age and gender effects on the pharmacokinetics of a single 500 milligram oral dose of levofloxacin in healthy subjects. AB - The influence of age and gender on the pharmacokinetics of levofloxacin in healthy subjects receiving a single oral 500-mg dose of levofloxacin was investigated in this parallel design study. Six young males (aged 18 to 40 years), six elderly males (aged > or = 65 years), six young females (aged 18 to 40 years), and six elderly females (aged > or = 65 years) were enrolled and completed the study. The study reveals that the bioavailability (rate and extent) of levofloxacin was not affected by either age or gender. In both age (young and elderly) and gender (male and female) groups of subjects, peak concentrations in plasma were reached at approximately 1.5 h after dosing; renal clearance of levofloxacin accounted for approximately 77% of total body clearance, and approximately 76% of the administered dose was recovered unchanged in urine over the 36 h of collection. The apparent differences in the calculated pharmacokinetic parameters for levofloxacin between the age groups (young versus elderly) and between the gender groups (males versus females) could be explained by differences in renal function among the subjects. A single dose of 500 mg of levofloxacin administered orally to both young and old, male and female healthy subjects was found to be safe and well tolerated. As the differences in levofloxacin kinetics between the young and the elderly or the males and the females are limited and are mainly related to the renal function of the subjects, dose adjustment based on age or gender alone is not necessary. PMID- 9210686 TI - Rifabutin absorption in the gut unaltered by concomitant administration of didanosine in AIDS patients. AB - Didanosine (ddI) is currently used in the management of patients infected by the human immunodeficiency virus. Rifabutin (RBT) is being extensively used for prophylaxis against Mycobacterium avium complex (MAC) infections. Due to its acid labile characteristics, ddI must be administered with a buffer. Recent reports have indicated that absorption of ketoconazole, ciprofloxacin, and dapsone, etc., in the gut is altered by concomitant ddI dosing. We have assessed whether concomitant dosing of ddI as antiretroviral therapy modifies RBT absorption in the gut, its steady-state pharmacokinetics, and/or safety in 15 patients with AIDS. Of the 15 patients enrolled, 12 completed the study and 3 receiving 600 mg of RBT with concomitant ddI administration withdrew prematurely from the study. Steady-state RBT pharmacokinetics were assessed on day 13 (ddI plus RBT) and day 16 (RBT alone). The ddI doses (adjusted for body weight) were 167 to 375 mg twice daily, while RBT was administered as a single 300- or 600-mg daily dose. No statistically significant (P > 0.05) differences were seen in RBT absorption parameter estimates between days 13 and 16: maximum concentration in plasma (Cmax; 511 +/- 341 ng/ml versus 525 +/- 254 ng/ml) and the time at which Cmax was observed (3.0 versus 2.5 h). The mean RBT estimates for area under the concentration-time curve from 0 to 24 h (AUC(0-tau)) (5,650 versus 5,023 ng x h/ml) and for oral clearance (1.28 versus 1.18 liter/h/kg) on both study days were also similar. Assessment based on urinary recovery of RBT (3.1 versus 3.7 mg) and its predominant deacetyl metabolite, LM565 (1.6 versus 1.4 mg), showed no apparent effect of ddI. The fraction of the RBT dose converted to LM565, as suggested by the ratio of AUC of the metabolite to AUC of the parent drug, was also unaltered (0.15 versus 0.12). A ratio analysis (day 13/day 16) of the RBT pharmacokinetic estimates showed that the 95% confidence intervals for all parameters were inclusive of one. Furthermore, the brief interruption of ddI therapy over this short study period at steady state produced no clinically significant changes in body weight, hematology, and renal and pancreatic functions. Therefore, concomitant administration of ddI appears not to affect RBT absorption in the gut and its disposition or safety in patients with AIDS. PMID- 9210687 TI - Pharmacokinetics of hyperimmune anti-human immunodeficiency virus immunoglobulin in persons with AIDS. AB - Hyperimmune anti-human immunodeficiency virus immunoglobulin (HIVIG) is an intravenous immunoglobulin prepared from HIV-infected asymptomatic donors with a CD4 cell count greater than 400 cells/microl and a high titer of antibody to HIV 1 p24 protein. Twelve persons with AIDS received four doses of HMG (two at 50 mg/kg of body weight and then two at 200 mg/kg) every 28 days. Pharmacokinetics were evaluated by measurement of anti-p24 antibody. HIVIG was well tolerated, and all participants completed the study. Three subjects who were not receiving Pneumocystis carinii pneumonia (PCP) prophylaxis developed PCP. The mean value for HIVIG clearance was 3.02 ml/kg/day at 50 mg/kg and 3.65 ml/kg/day at 200 mg/kg (P = 0.027); the mean trough antibody titers (reciprocal units) were 1,442 and 4,428, respectively. This study indicates that high titers of anti-p24 antibody can be maintained with a monthly administration schedule of HIVIG and that short-term safety is acceptable. Comparisons to evaluate the therapeutic potential of HIVIG are justified. PMID- 9210688 TI - Effect of granulocyte colony-stimulating factor on the candidacidal activity of polymorphonuclear neutrophils and their collaboration with fluconazole. AB - The effect of granulocyte colony-stimulating factor (GCSF) treatment of polymorphonuclear neutrophils (PMN) in vitro was studied with respect to their candidacidal activity. The candidacidal activity of PMN was found to be significantly increased when they were pretreated with GCSF. Fluconazole (1 microg/ml) was found to be highly fungistatic (90%) for Candida albicans Sh27 and collaborated with PMN for significantly increased killing. Collaborative killing by PMN significantly increased when they were treated with GCSF before and after fungal exposure. The enhancing activities of GCSF required optimization of the GCSF dose and were thus inoculum and strain dependent. PMID- 9210689 TI - Comparison of cefdinir and cefaclor in treatment of community-acquired pneumonia. AB - Six hundred ninety patients were enrolled in a multicenter, randomized, double blind trial comparing the efficacy and safety of cefdinir with those of cefaclor in the treatment of community-acquired pneumonia. Patients received either 10 days of treatment with cefdinir (n = 347) at 300 mg twice daily or 10 days of treatment with cefaclor (n = 343) at 500 mg three times daily. Microbiological assessments were performed on sputum specimens obtained at admission and at the two posttherapy visits, if available. Respiratory tract pathogens were isolated from 538 (78%) of 690 patient admission sputum specimens, with the predominant pathogens being Haemophilus parainfluenzae, Haemophilus influenzae, Streptococcus pneumoniae, and Staphylococcus aureus. The microbiological eradication rates at the test-of-cure visit were 92% (238 of 260 pathogens) and 93% (245 of 264 pathogens) for the evaluable patients treated with cefdinir and cefaclor, respectively. A satisfactory clinical response (cure plus improvement) was achieved in 89% (166 of 187) and 86% (160 of 186) of the evaluable patients treated with cefdinir and cefaclor, respectively. Except for the incidence of diarrhea, adverse event rates while on treatment were equivalent between the two treatment groups. Diarrhea incidence during therapy was higher for patients treated with cefdinir (13.7%) than for patients treated with cefaclor (5.3%). These results indicate that cefdinir is effective and safe in the treatment of patients with pneumonia. PMID- 9210690 TI - Prospective randomized comparison of cefodizime versus cefuroxime for perioperative prophylaxis in patients undergoing coronary artery bypass grafting. AB - The effects of cefodizime and cefuroxime on neutrophil phagocytosis and reactive oxygen production in 54 patients undergoing elective coronary artery bypass grafting were studied. Both drugs were administered twice at a dosage of 40 mg/kg of body weight (pre- and intraoperative). Phagocytic capacity was assessed by measuring the uptake of fluorescein isothiocyanate-labeled Escherichia coli and Staphylococcus aureus by flow cytometry. Reactive oxygen generation after phagocytosis was estimated by determining the amount of dihydrorhodamine 123 converted to rhodamine 123 intracellularly. In both groups the mean phagocytic ability for E. coli and S. aureus decreased during surgery (-21 and -8%, respectively, for the cefodizime group and -39 and -38%, respectively, for the cefuroxime group; P < 0.05 for all). In the cefodizime group a normalization of mean E. coli and S. aureus neutrophil phagocytosis was seen on day 5 (+9 and -4% compared to preoperative values; P > 0.35 for both), whereas in cefuroxime treated patients phagocytic ability remained depressed (-37 and -31%; P < 0.04 for both). In both groups mean neutrophil reactive oxygen intermediate (ROI) production after E. coli and S. aureus phagocytosis increased during cardiopulmonary bypass (+44 and +83%, respectively, in the cefodizime group and +58 and +73%, respectively, in the cefuroxime group; P < 0.05 for all). One day after surgery E. coli- and S. aureus-driven neutrophil ROI production was not different from the preoperative values (-2 and +12%, respectively, for the cefodizime group and +7 and +15%, respectively, for the cefuroxime group; P > 0.15 for all). Postoperative serum levels of the C-reactive protein on days 2 and 7 were lower in cefodizime-treated patients (19 +/- 6 and 4 +/- 2 mg/liter versus 23 +/- 6 and 11 +/- 5 mg/liter; P < 0.05 for both). In addition to cefodizime's antimicrobial activity during perioperative prophylaxis, its use in coronary artery bypass grafting can prevent procedure-related prolonged postoperative neutrophil phagocytosis impairment. PMID- 9210691 TI - Effect of severity of meningitis on fungicidal activity of flucytosine combined with fluconazole in a murine model of cryptococcal meningitis. AB - We studied the effect of the severity of meningitis on the response to therapy with fluconazole and flucytosine in a murine model of cryptococcal meningitis. Meningitis was established by intracerebral injection of Cryptococcus neoformans. The severity of meningitis was varied by delaying the onset of treatment from 3 to 7 days. Animals were sacrificed after 14 days of treatment, and the numbers of C. neoformans per gram of brain tissue were quantified. The range of effective dose combinations of fluconazole and flucytosine became progressively reduced as the severity of meningitis increased. The magnitude of treatment effect, as measured by the numbers of CFU/gram of brain tissue, was also reduced with increasing severity of meningitis. In this model, as the severity of meningitis increases, higher doses of fluconazole are required to achieve equivalent levels of activity. The combination of fluconazole and flucytosine appears to have the most-potent antifungal effects. This is most readily observed in animals with more-severe meningitis. PMID- 9210693 TI - Antibiotic resistance among enterococci isolated from clinical specimens between 1953 and 1954. AB - Two hundred twenty group D streptococci isolated from 1953 to 1954 from patients in the Washington, D.C., area were characterized. All were susceptible to ampicillin, vancomycin, and gentamicin; none produced beta-lactamase activity. High-level resistance to streptomycin was expressed by 117 strains, and 2 strains were resistant to >8 microg of chloramphenicol per ml. Three isolates were resistant to both erythromycin and lincomycin, and DNA from these hybridized to an ermAM probe. Of 118 strains resistant to tetracycline and minocycline, genomic DNA from 63 was examined for homology to tet(M), tet(O), and tet(S). DNA from 20 strains hybridized to tet(M), DNA from 37 strains hybridized to tet(S), and DNA from none of the strains hybridized to tet(O). PMID- 9210692 TI - In vitro activity of BAY 12-8039, a novel 8-methoxyquinolone, compared to activities of six fluoroquinolones against Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis. AB - The in vitro activity of a novel 8-methoxyquinolone, BAY 12-8039, against recent clinical isolates of Streptococcus pneumoniae (n = 404), Haemophilus influenzae (n = 330), and Moraxella catarrhalis (n = 250) was evaluated. Activity was compared to those of six other fluoroquinolones: ciprofloxacin, clinafloxacin, levofloxacin, ofloxacin, sparfloxacin and trovafloxacin. BAY 12-8039 and clinafloxacin had the highest levels of activity against S. pneumoniae, both with a MIC at which 90% of the isolates were inhibited (MIC90) of 0.06 microg/ml. Trovafloxacin and sparfloxacin were the next most active agents versus S. pneumoniae (MIC90s = 0.12 microg/ml). No differences in activity against penicillin-susceptible, -intermediate, or -resistant strains of S. pneumoniae were noted for any of the fluoroquinolones tested. MIC90s for the seven fluoroquinolones ranged from 0.008 to 0.06 microg/ml versus H. influenzae and from 0.008 to 0.12 microg/ml for M. catarrhalis. The MICs for two strains of S. pneumoniae and one strain of H. influenzae were noted to be higher than those for the general population of organisms for all of the fluoroquinolones tested. Finally, the activity of BAY 12-8039 versus S. pneumoniae was found to be diminished when MIC determinations were performed with incubation of agar dilution plates or broth microdilution trays in 5 to 7% CO2 versus ambient air. PMID- 9210694 TI - Molecular analysis of katG gene mutations in strains of Mycobacterium tuberculosis complex from Africa. AB - A sample of 124 isoniazid (INH)-resistant and 88 susceptible strains of Mycobacterium tuberculosis complex from south, central, and west Africa was analyzed by direct sequence analysis and PCR-restriction fragment length polymorphism analysis of their catalase-peroxidase (katG) genes. Point mutations at codon 315 were found in the genomes of 64% of INH-resistant strains, but no complete deletions were identified. Mutations at codon 463 were independent of INH resistance and were linked to the geographic origins of the strains. PMID- 9210695 TI - Cross-resistance associated with development of resistance to isometamidium in a clone of Trypanosoma congolense. AB - Resistance to isometamidium was increased 94-fold in a clone of Trypanosoma congolense (clone IL 1180) by repeated subcurative treatment of infected mice for 11 months. This was associated with 3.4-, 33-, and 4.2-fold increases in resistance to diminazene, homidium, and quinapyramine, respectively. Both T. congolense IL 1180 and the resistant derivative were able to undergo cyclical development in Glossina morsitans centralis tsetse flies, producing hypopharyngeal infection rates of 40.0 and 39.8%, respectively. PMID- 9210696 TI - In vitro synergy testing of clarithromycin and 14-hydroxyclarithromycin with amoxicillin or bismuth subsalicylate against Helicobacter pylori. AB - The activity of clarithromycin-14-hydroxyclarithromycin (2:1 ratio) and bismuth subsalicylate or amoxicillin against Helicobacter pylori was determined by the checkerboard technique in vitro. Clarithromycin-14-hydroxyclarithromycin and amoxicillin resulted in additive effects in 7 of 22 isolates, compared to 14 of 22 isolates when bismuth subsalicylate was substituted for amoxicillin. Synergy was not demonstrated and is probably not responsible for the clinical success of treatment combinations containing clarithromycin. PMID- 9210697 TI - In vivo selection of Klebsiella pneumoniae strains with enhanced quinolone resistance during fluoroquinolone treatment of urinary tract infections. AB - We report two cases of failure of fluoroquinolone treatment of urinary tract infections with Klebsiella pneumoniae strains harboring quinolone resistance associated alterations in GyrA and ParC and in vivo selection of posttreatment isolates with enhanced fluoroquinolone resistance. Active efflux leading to decreased accumulation of a drug enhanced fluoroquinolone resistance in one posttreatment isolate, and an additional mutation in parC resulting in an additional amino acid change in ParC was associated with increased resistance in the other. PMID- 9210698 TI - In vitro activity of a new pneumocandin antifungal, L-743,872, against azole susceptible and -resistant Candida species. AB - The in vitro activity of a new pneumocandin, L-743,872, was evaluated with 108 strains of Candida and compared with the activities of various antifungals. L 743,872 demonstrated the best activity against azole-susceptible and -resistant strains of C. albicans, C. glabrata, C. tropicalis, C. parapsilosis, and C. kefyr and less activity against C. krusei, C. lusitaniae, and C. guilliermondii. PMID- 9210699 TI - Small, anionic, and charge-neutralizing propeptide fragments of zymogens are antimicrobial. AB - Some inactive precursor proteins, or zymogens, contain small, amino terminus, homopolymeric regions of Asp that neutralize the cationic charge of the active protein during synthesis. After posttranslational cleavage, the anionic propeptide fragment may exhibit antimicrobial activity. To demonstrate this, ovine trypsinogen activation peptide, and frog (Xenopus laevis) PYL activation peptide, both containing homopolymeric regions of Asp, were synthesized and tested against previously described surfactant-associated anionic peptide. Peptides inhibited the growth of both gram-negative (MIC, 0.08 to 3.00 mM) and gram-positive (MIC, 0.94 to 2.67 mM) bacteria. Small, anionic, and charge neutralizing propeptide fragments of zymogens form a new class of host-derived antimicrobial peptides important in innate defense. PMID- 9210700 TI - Regimens to treat lepromatous leprosy. PMID- 9210701 TI - Failure of short antimicrobial treatments for human brucellosis. PMID- 9210702 TI - Continuous infusion of 5-fluorouracil and low dose cisplatin infusion for the treatment of advanced and recurrent gastric adenocarcinoma. AB - BACKGROUND: Several chemotherapy studies have suggested that continuous infusion of 5-fluorouracil (5-FU) is more effective than bolus 5-FU. In addition, 5-FU and cis-Diamminedichloroplatinum-II (cisplatin) in combination have been shown to have synergistic cytotoxicity against several human neoplasms. In this study, the authors evaluated the efficacy and toxicity of continuous infusion of 5-FU and low dose cisplatin infusion (FP therapy) in the treatment of advanced and recurrent gastric adenocarcinoma. The relationship between the response to FP therapy and several factors was also examined. METHODS: A total of 26 patients fulfilling standard eligibility criteria were enrolled in the trial. FP therapy consisted of 5-FU (350 mg/m2/day every day by continuous infusion) and cisplatin (7.5 mg/m2/day in 100 mL of normal saline infused over 1 hour on Days 1-5 every week) for 4 weeks. RESULTS: A complete response was observed in 2 cases and a partial response in 11 cases, for an overall response rate of 50%. Patients with good performance status (PS) (0-1) and differentiated histologic type showed higher response rates (50.0% and 63.6%, respectively) than patients with poor PS (2 or 3) and undifferentiated histologic type (28.6% and 35.3%, respectively), although there were no significant differences. Patients with low serum levels of immunosuppressive acidic protein (IAP) showed a significantly higher response rate (71.4%) than those with high IAP levels (0%). Toxic effects included leukopenia, thrombocytopenia, nausea, and vomiting; these were not life threatening and did not require treatment interruption. CONCLUSIONS: FP therapy is a promising regimen for patients with advanced and recurrent gastric adenocarcinoma. Serum levels of IAP may predict chemosensitivity. PMID- 9210704 TI - Pilot study--cimetidine enhances lymphocyte infiltration of human colorectal carcinoma: results of a small randomized control trial. AB - BACKGROUND: Cimetidine preserves postoperative immune function and inhibits the growth of some cancers. In this study, the effect of cimetidine on the local immune response to colorectal carcinoma was investigated. METHODS: Forty-two patients scheduled for elective resection of colorectal carcinoma were randomized either to receive cimetidine for 1 week perioperatively or to act as controls. A lymphocyte density of 50 cells per high-power field (approximately 50% of the tumor/tissue interface) was considered a positive response. Patient survival was determined by Kaplan-Meier life table analysis. The effects of histamine and cimetidine on normal subject lymphocyte function was determined in a mitogen stimulated proliferation assay. RESULTS: A positive lymphocyte response was observed in 5 of 24 control carcinoma patients (21%) and 10 of 18 cimetidine treated carcinoma patients (56%) (P = 0.03). The presence of a lymphocyte response correlated with a better survival (P = 0.02). Histamine had an inhibitory effect on lymphocyte proliferation with a median effective dose of 5 x 10(-7) M. Cimetidine antagonized this effect with a negative logarithm of the cimetidine molar concentration required to reduce the effect of histamine in half of 6.55. CONCLUSIONS: Histamine inhibits normal lymphocyte function, antagonized by cimetidine at a histamine type 2 receptor. Cimetidine increases lymphocyte infiltration of primary colorectal carcinoma, possibly by overcoming the immunosuppressive effects of high local histamine concentrations. The presence of a local lymphocyte response correlates with an improved 3-year survival. PMID- 9210703 TI - Five-year results of the treatment of 23 patients with immunoproliferative small intestinal disease: a Turkish experience. AB - BACKGROUND: Currently, there is no agreement regarding optimal treatment strategies for immunoproliferative small intestinal disease (IPSID). In this article, the authors report the treatment outcomes of a group of 23 Turkish patients with IPSID. METHODS: Between December 1988 and July 1993, 23 consecutive patients with IPSID, including 5 with secretory type, were included in the study. Seven patients with Stage A disease (according to the criteria of Galien et al.) received tetracycline (1 g/day, orally) for a median duration of 7 months (range, 6-11 months) initially, whereas the remaining patients (9 Stage B patients and 7 Stage C patients) received combination chemotherapy (cyclophosphamide, vincristine, procarbazine, and prednisolone [COPP regimen]) followed by tetracycline at a dose of 1 g/day for 6 more months in patients with complete response (CR) after the COPP regimen. RESULTS: The median follow-up was 68 months (range, 38-89 months). As first-line therapy in Stage A patients, tetracycline yielded a 71% CR and 43% disease free survival (DFS) rate. Eleven of 16 patients (69%) with Stage B or C disease who received the COPP regimen achieved CR and only 2 patients had a recurrence (DFS rate of 56%). The 5-year overall survival (OAS) rate for the entire group was 70%, and the 5-year DFS rate for patients with CR was 75%. However, the median OAS for 3 patients with immunoblastic lymphoma was only 7 months. CONCLUSIONS: The COPP regimen, with its acceptable toxicity, appears to be a good alternative as a first-line treatment for patients with Stage B or C IPSID with low grade lymphoma whereas tetracycline appears to be the initial treatment of choice for patients with Stage A disease. PMID- 9210705 TI - In situ simultaneous detection of hepatitis C virus RNA and hepatitis C virus related antigens in hepatocellular carcinoma. AB - BACKGROUND: The overwhelming evidence that chronic infection with the hepatitis C virus (HCV) is an important cause of hepatocellular carcinoma (HCC) is based on epidemiologic, case-control, and cohort studies as well as laboratory investigations. To address better the pathogenesis of HCV infection at the single cell level, the authors developed a specific reproducible method for the simultaneous detection of HCV specific sequences and antigens in liver tissue, using a combination of nonradioactive in situ hybridization and immunohistochemistry. METHODS: After immunohistochemical staining of the liver sections for E2/NS-1, C22-3, C33c, C100-3, and NS-5 antigens with immunogold silver technique, in situ hybridization was performed on the same sections using digoxigenin-labeled HCV 5' NonCoding specific probes. The hybridization signal was detected by an antidigoxigenin, Fab fragment-alkaline phosphatase conjugate. This simultaneous detection permitted the subcellular localization of HCV RNA and antigens with excellent preservation of tissue morphology and absence of background staining. In addition, the types and percentages of cells harboring HCV in tissue could be determined. RESULTS: The in situ detection of HCV showed positive signals in both cancerous and noncancerous areas of liver tissue in six of six HCV-infected patients with HCC and in none of four controls, including three HCV negative HCC patients and one patient with epithelioid hemangioendothelioma. Two classes of infected cells were distinguished throughout the liver: (1) cells containing large amounts of negative-stranded HCV RNA, which were probably undergoing active viral replication; and (2) cells displaying positive-stranded HCV RNA only, with unpredictable levels of viral replication. Both types expressed core, envelope, and NS-3, -4, and -5 proteins. HCV RNA and antigens were exclusively cytoplasmic. Detection of viral proteins was highly predictive of the presence of large amounts of HCV RNA in the same cell. Fewer HCV positive cells were consistently demonstrated in the cancerous area. CONCLUSIONS: These findings support the contention that HCV infects hepatocytes and replicates in them, even after their malignant transformation. PMID- 9210706 TI - CD44 and its v6 spliced variant in lung tumors: a role in histogenesis? AB - BACKGROUND: CD44 is a polymorphic family of cell surface glycoproteins with a variety of functions including participation in cell adhesion and migration as well as modulation of cell-matrix interactions. Expression of the standard form of CD44 (CD44s) and its variant isoforms has been shown in both normal and neoplastic tissue and holds promise as a prognostic indicator. METHODS: The authors investigated the expression of CD44s and its v6 isoform (CD44v6) immunohistochemically in 7 fetal lungs (gestational age between 11-36 weeks) and in 80 lung tumors of various histologic types, degrees of differentiation, and clinical stages. RESULTS: In the fetal lung, CD44v6 was expressed as membranous and luminal staining of epithelial cells throughout gestation and basal staining of bronchial epithelium late in gestation. Nonneoplastic adult lung showed CD44v6 expression that was restricted to epithelial cells with membranous staining of basal bronchial cells and squamous metaplasia as well as basolateral membranous staining of type 2 pneumocytes. CD44s showed similar but less intense staining and was in addition present on lymphocytes and macrophages. Tumorlets and neuroepithelial bodies were CD44v6 negative. Nearly all squamous cell carcinomas (97%) were positive for CD44v6 with patterns similar to squamous metaplasia and with more intense staining at the periphery of tumor nests. Most adenocarcinomas (90%) were CD44v6 negative whereas most bronchioloalveolar cell carcinomas (71%) were CD44v6 positive with patterns similar to that in type 2 pneumocytes. Most large cell carcinomas (71%), carcinoid tumors (67%), and all small cell carcinomas were CD44v6 negative. CD44v6 expression did not correlate with clinical stage. CD44v6 expression in lymph node metastases was identical to that of the primary tumor. CONCLUSIONS: The results of the current study show that CD44v6 is localized differently in fetal and adult lung, suggesting a difference in function. In the fetal lung, it may modulate growth factors important in morphogenesis and maturation. In the adult nonneoplastic lung, CD44v6 is associated with stem cells, namely basal cells and type 2 pneumocytes, and may act to anchor these cells to the matrix and be important in migration during repair or neoplasia. In addition, CD44v6 expression is maintained throughout tumorigenesis in squamous cell carcinoma and bronchioloalveolar cell carcinoma, suggesting a histogenetic relationship between the stem cells and the respective tumors. Conversely, most neuroendocrine tumors and the cells of the dispersive neuroendocrine system do not express CD44v6, implying a separate histogenetic lineage in these tumors. PMID- 9210707 TI - A phase III randomized study of interleukin-2 lymphokine-activated killer cell immunotherapy combined with chemotherapy or radiotherapy after curative or noncurative resection of primary lung carcinoma. AB - BACKGROUND: Adoptive immunotherapy with interleukin-2 (IL-2) and lymphokine activated killer (LAK) cells has resulted in response among some patients with advanced malignant disease. However, the relative therapeutic benefit of adoptive immunotherapy as an adjuvant to surgery has not been determined. METHODS: A Phase III prospective randomized controlled study of adjuvant immunotherapy combined with chemotherapy or radiotherapy was conducted for 174 primary lung carcinoma patients postsurgically. After a pathologic examination of resected tissues, patients were divided into curative and noncurative cases and randomized to receive either combined immunotherapy (Group A) or control standard therapy (Group B). Patients who had undergone curative resection of lung carcinoma were stratified according to the stages and histologic types of their disease, and those who had undergone noncurative resection were stratified according to the causes of noncurative resection. The patients of Group A received IL-2 and LAK cells after either chemotherapy or radiotherapy, and those in Group B received either no adjuvant therapy (curative cases) or radiotherapy or chemotherapy alone (noncurative cases), according to the causes of noncurative resection. RESULTS: The 5- and 9-year survival rates of the Group A patients were 54.4% and 52.0%, respectively, and those of the Group B patients were 33.4% and 24.2%, respectively. The difference was statistically significant (P < 0.001, according to the generalized Wilcoxon's test and the Cox-Mantel test). The difference was also statistically significant in the curative cases (65.5% for Group A vs. 40.6% for Group B; 5-year survival rate, P < 0.01), noncurative cases (43.0% for A vs. 20.8% for B; P < 0.01), adenocarcinoma cases (47.5% for A vs. 23.0% for B; P < 0.05), and squamous cell carcinoma cases (62.1% for A vs. 34.8% for B; P < 0.01). CONCLUSIONS: Adoptive immunotherapy with IL-2 and LAK cells combined with chemotherapy or radiotherapy improved the survival of patients after surgical resection of primary lung carcinoma. A multi-institutional group study should be carried out to verify the significance of this study. PMID- 9210708 TI - Chondrosarcoma of small bones of the hands and feet. AB - BACKGROUND: Cartilaginous tumors of the hands and feet are not uncommon. Most are enchondromas, but they tend to show high cellularity, enlargement of nuclei, and many double-nucleated cells. Hence, differentiation between a benign lesion and chondrosarcoma may be difficult. METHODS: The files of patients treated at the Mayo Clinic and the consultation files were reviewed for examples of chondrosarcoma of small bones of the hands and feet. Histologic features and clinical charts were reviewed in all cases, and radiographs were reviewed in 111 cases. RESULTS: Seventy-five lesions involved the feet and 88 involved the hands. Bones of the fifth finger and the calcaneus were the most common sites of involvement. Nineteen tumors were secondary. Of the 104 intramedullary lesions studied radiologically, 96 showed cortical destruction, 83 a soft tissue mass, and 52 a permeative lytic pattern. Histologically, soft tissue extension and permeation of preexisting bone indicated malignant disease. One hundred sixteen tumors were Grade 1, 44 were Grade 2, and 3 were Grade 3. Of the 12 patients with distant metastasis from chondrosarcoma, 7 died of disease. Chondrosarcomas of the calcaneus and the talus were more likely to metastasize. CONCLUSIONS: Chondrosarcoma of small bones of the hands and feet has the potential to be fatal. PMID- 9210709 TI - Metastatic melanoma of unknown primary origin shows prognostic similarities to regional metastatic melanoma: recommendations for initial staging examinations. AB - BACKGROUND: Metastatic melanoma of unknown primary origin accounts for approximately 2-6% of all melanoma cases. The prognostic significance of this diagnosis is still controversial. METHODS: Of 3258 patients with malignant melanoma recorded during the period 1976-1996, 2.3% had metastases of unknown primary origin. Anatomic distribution, clinical stage, and survival probabilities were evaluated. RESULTS: Thirty patients were classified as having cutaneous or subcutaneous in-transit metastases, and they showed a 5-year survival rate of 83%. Thirty-seven patients were classified as having lymph node metastasis, and their 5-year survival rate was 50%. Disseminated disease was diagnosed in only 8 patients, who had a median survival of 6 months. Comparison of survival probabilities for patients with in-transit metastases and unknown primary tumors with the probabilities for those with cutaneous primary tumors revealed a significant advantage for the former group. No significant differences were found for patients with lymph node metastasis when those with unknown primary tumors were compared with those who had cutaneous melanomas with regional lymph node metastasis. CONCLUSIONS: The clinical disease course of patients with metastatic melanoma of unknown primary origin is similar to that of patients with primary cutaneous melanoma when the same clinical stages of the disease are compared. Based on the assumption that the majority of regional metastases develop from completely regressed primary cutaneous melanoma, recommendations for initial staging examinations in patients with unknown primary tumors are given in this article. PMID- 9210710 TI - Automated analysis of mammographic densities and breast carcinoma risk. AB - BACKGROUND: There is considerable evidence that one of the strongest risk factors for breast carcinoma can be assessed from the mammographic appearance of the breast. However, the magnitude of the risk factor and the reliability of the prediction depend on the method of classification. Subjective classification requires specialized observer training and suffers from inter- and intraobserver variability. Furthermore, the categoric scales make it difficult to distinguish small differences in mammographic appearance. To address these limitations, automated analysis techniques that characterize mammographic density on a continuous scale have been considered, but as yet, these have been evaluated only for their ability to reproduce subjective classifications of mammographic parenchyma. METHODS: In this study, using a nested case-control design, the authors evaluated the direct association between breast carcinoma risk and quantitative image features derived from automated analysis of digitized film mammograms. Two parameters, one describing the distribution of breast tissue density as reflected by brightness of the mammogram (regional skewness) and the other characterizing texture (fractal dimension), were calculated for images from 708 subjects identified from the Canadian National Breast Screening Study. RESULTS: These parameters were evaluated for their ability to distinguish cases (those women who developed breast carcinoma) from controls. It was found that both the skewness and fractal parameters were significantly related to risk of developing breast carcinoma. CONCLUSIONS: Although the relative risk estimates were moderate (typically > 2.0) and less than those from subjective classification or for an interactive computer method the authors have previously described, they are comparable to other risk factors for the disease. The observer independence and reproducibility of the automated methods may facilitate their more widespread use. PMID- 9210711 TI - The results of intraoperative consultations in 181 ductal carcinomas in situ of the breast. AB - BACKGROUND: The utility of frozen section (FS) examination in the intraoperative management of breast lesions is well established. The accuracy of FS in the diagnosis of borderline noninvasive or preinvasive breast lesions is uncertain. METHODS: The authors retrospectively reviewed the results of intraoperative consultations/frozen section examinations of 181 ductal carcinomas in situ (DCIS) of the breast. Various clinical and pathologic factors were analyzed and correlated with FS diagnosis. RESULTS: FS examination was performed on 153 cases (85%) and only macroscopic examination on 28 cases (15%). FS diagnoses were as follows: DCIS in 76 cases (50%), atypical ductal hyperplasia/suspicious for DCIS in 8 cases (5%), benign in 55 cases (36%), deferred in 13 cases (8%), and invasive carcinoma in 1 case. FS accuracy, false-negative rate, and false positive rate were 55%, 36%, and 0.6%, respectively. Sampling error was the main reason for the low detection rate, and technical inadequacy was a major factor contributing to interpretive problems. In multivariate regression analysis, FS accuracy was significantly associated with the clinical presentation of a palpable mass (odds ratio [OR] = 4.16, 95% confidence interval [CI]: 2.04-8.45), the macroscopic finding of a mass (OR = 3.03, 95% CI: 1.45-6.67), and necrosis (OR = 3.13, 95% CI: 1.4-6.67). CONCLUSIONS: The authors concluded that the accuracy of FS diagnosis of DCIS was low, mainly due to sampling error. In general, FS examination should not be performed when no lesion/mass is identified by macroscopic examination. PMID- 9210712 TI - Prognosis among African-American women and white women with lymph node negative breast carcinoma: findings from two randomized clinical trials of the National Surgical Adjuvant Breast and Bowel Project (NSABP). AB - BACKGROUND: A disparity in breast carcinoma survival between African-American and white women has been noted over the past several decades. A major factor implicated in this disparity is stage of disease at diagnosis. In this study, survival and related endpoints were examined among African-American women and white women with lymph node negative breast carcinoma who participated in two randomized clinical trials of the National Surgical Adjuvant Breast and Bowel Project (NSABP). METHODS: Patients from two studies, one conducted among patients with estrogen receptor (ER) negative tumors and the other among patients with ER positive tumors, were included. Study goals were to determine whether African Americans and whites had comparable outcomes, accounting for ER status and differences in patient characteristics at diagnosis, and to determine whether treatment response was similar for African-Americans and whites. RESULTS: Five year survival rates were 83% for African-Americans and 85% for whites among ER negative patients, and 93% for African-Americans and 92% for whites among ER positive patients. Rates of disease free survival (DFS) (i.e., time to disease recurrence, second primary cancer, or death) were 71% for African-Americans and 74% for whites at 5 years among ER negative patients, and 81% for African Americans and 80% for whites among ER positive patients. African-Americans tended to have less favorable baseline prognostic characteristics. Adjusted relative risk (RR) estimates indicated similar prognosis for African-Americans compared with whites for mortality (African-American/white RR = 1.02 with 95% confidence interval [CI], 0.66-1.56 among ER negative patients; RR = 1.14 with 95% CI, 0.84 1.54 among ER positive patients) and DFS (RR = 0.98 with 95% CI, 0.70-1.37 for ER negative patients; RR = 0.96 with 95% CI, 0.75-1.22 for ER positive patients). Estimated percent reductions in DFS events for patients receiving adjuvant therapy were 32% for ER negative African-Americans, 36% for ER negative whites, 20% for ER positive African-Americans, and 39% for ER positive whites. CONCLUSIONS: African-American and white patients with localized breast carcinoma had similar outcomes and benefited equally from systemic therapy. These results suggest that early detection and appropriate therapy among African-American patients could result in a reduction in the current disparity in breast carcinoma mortality between African-Americans and whites. PMID- 9210713 TI - Lymphoepithelioma-like carcinoma of the uterine cervix: association with Epstein Barr virus and human papillomavirus. AB - BACKGROUND: The presence of Epstein-Barr virus (EBV) has not been documented in previous reports of lymphoepithelioma-like carcinoma (LELC) of the uterine cervix by either polymerase chain reaction or in situ hybridization, and the histogenesis of the tumor remains unknown. Additionally, a relationship between human papillomavirus (HPV) and cervical LELC also has not been reported. METHODS: In this article, the authors describe the clinical and histopathologic findings for 15 patients with cervical carcinoma that had a histologic pattern of LELC. The polymerase chain reaction detected the presence of EBV and HPV DNA sequences in cervical LELC. RESULTS: All 15 tumors showed a typical syncytial growth pattern of undifferentiated cells with prominent lymphocytic infiltration. The detection rate of the EBV gene sequence in tissue samples from patients with LELC was more frequent than that in control patients with squamous cell carcinoma of the cervix (11 of 15 patients, 73.3%, vs. 4 of 15 patients, 26.7%; P = 0.01). However, the detection rate of HPV-16 and HPV-18 DNA was significantly lower in patients with LELC tumors than in patients with cervical squamous cell carcinoma (3 of 15 patients, 20.0%, vs. 12 of 15 patients, 80.0%; P = 0.001). After a median follow-up of 3.9 years (range, 1.8-5.3 years), the 15 patients showed no evidence of disease or metastasis after radical hysterectomy or radiotherapy. CONCLUSIONS: The finding of EBV associations in cervical LELC supports the hypothesis that EBV may be involved in the pathogenesis of tumors that arise in the cervix. It is possible that cervical LELC may follow a different pathway in the pathogenesis of LELC in Asian women as compared with the more common forms of squamous cell carcinoma. PMID- 9210714 TI - Vascular endothelial growth factor expression in early stage ovarian carcinoma. AB - BACKGROUND: Tumor angiogenesis is essential for solid tumor growth. Yet, the importance of any particular factor in neoplastic proliferation is poorly defined. This study examines the clinical significance of increased expression of one of the angiogenic factors, vascular endothelial growth factor (VEGF), in early stage ovarian carcinoma. METHODS: Tumor specimens from 68 patients with International Federation of Gynecology and Obstetrics Stage I and II ovarian carcinoma were evaluated for VEGF expression. Antisense and corresponding sense (control) RNA probes were transcribed from the pCRII construct (Invitrogen, San Diego, CA), which contained human VEGF cDNA. The antisense probe was designed to include a highly conserved region of the VEGF coding sequence and thus detect all known variants. After in situ hybridization, sections were assessed for overexpression of VEGF. RESULTS: Twenty-nine of the tumor samples overexpressed VEGF, whereas 39 specimens did not. In patients whose tumors demonstrated elevated VEGF expression, 25% were without evidence of disease recurrence at last follow-up. In contrast, 75% of the patients whose tumors did not overexpress VEGF were without evidence of disease at last follow-up (P < 0.001). Median disease free survival for the VEGF positive group was 22 months, compared with > 108 months for the VEGF negative group (P < 0.001). When borderline tumors were excluded from the survival analysis, median disease free survival for the VEGF positive group was 18 months, compared with >120 months for the VEGF negative group (P < 0.001). Other possible prognostic variables had minimal impact on survival; these included age, stage, grade, cytology, and tumor size (P > 0.05). Assignment to a high risk group, as defined by the Gynecologic Oncology Group of the National Cancer Institute, was somewhat predictive of a shorter relapse free interval (P = 0.056). In a multivariate analysis, however, only elevated VEGF expression was associated with poorer survival. CONCLUSIONS: In this analysis, patients with early stage ovarian carcinoma with increased VEGF expression had a poorer prognosis. Further study of VEGF may ultimately lead to identification of patients with high risk lesions whose tumor biology portends a worse prognosis and who therefore may benefit from aggressive adjuvant therapy. PMID- 9210715 TI - Comparison of serum prostate specific membrane antigen, prostate specific antigen, and free prostate specific antigen levels in radical prostatectomy patients. AB - BACKGROUND: Higher preoperative prostate specific antigen (PSA) levels are associated with higher pathologic stage and grade in patients undergoing radical prostatectomy (RP). In earlier studies, serum prostate specific membrane antigen (PSMA) elevations were associated with clinical progression and hormone refractory carcinoma. The goal of this study was to evaluate the serum markers PSMA, free PSA (FPSA), free:total PSA ratio (F:TPSA), and total PSA (PSA) in men undergoing RP. METHODS: Serum was obtained from 63 patients undergoing RP for clinically localized (T1c, T2) prostate carcinoma. Serum PSA and FPSA were determined by Hybritech Tandem-E(R) and Tandem-R(R), respectively, and PSMA was determined by Western blot analysis. Serum values for these markers were compared with the pathologic stage, surgical margin status, Gleason sum, prostate size (as calculated via reconstruction and transrectal ultrasound), tumor size based on pathologic assessment of the whole mount, and World Health Organization (WHO) grade of the prostatectomy specimen. Markers were also compared against demographic information and the patients' age and race. RESULTS: There was a weak correlation between serum PSA and positive surgical margins, higher Gleason sum, and WHO grade (P < 0.05). Receiver operating characteristic curve (ROC) analysis comparing sensitivity and specificity of the markers to positive and negative margins as well as seminal vesicle invasion demonstrated PSA and FPSA predictive ability for seminal vesicle invasion. The area under the curve for PSA and FPSA in this case was 0.7318 and 0.7432, respectively. There was also a weak correlation between the FPSA level and margins, with a low ROC area under the curve of 0.6789. The FPSA cannot distinguish the more advanced stage of disease. There was no significant correlation between F:TPSA and PSMA with regard to the study variables in predicting organ confinement. High PSMA levels only correlated with higher stage and were maximal in pT4a classified disease. CONCLUSIONS: Higher PSA and FPSA levels are likely to be associated with more locally advanced disease. Total PSA was the best marker. However, the cutoff values necessary for significant accuracy between PSA and FPSA are not of clinical usefulness due to the lack of specificity and sensitivity of the markers at those cutoffs. F:TPSA and PSMA levels as currently measured are of limited value in discriminating more aggressive disease in patients with clinically localized CaP. PMID- 9210716 TI - Radical chemoradiotherapy for elderly patients with bladder carcinoma invading muscle. AB - BACKGROUND: Chemoradiotherapy is becoming an alternative to radical cystectomy among patients with bladder carcinoma invading muscle. In 1988, the authors began a protocol with methotrexate, vinblastine, doxorubicin, and cisplatin (M-VAC regimen) and radiotherapy for these patients. Traditionally, age has been considered a determinant factor thereby excluding the older patients from the oncologic protocols that are considered to be more aggressive. The authors analyzed 20 patients (age > 70 years) who were treated during this period with the same protocol as the authors' other patients. METHODS: The study included 20 patients (age range, 70-78 years; median age, 74 years) including 4 patients with T2 disease, 9 with T3 disease, and 7 with T4 disease. All patients had a Karnofsky performance status of > 60. Treatment protocol included cytoreductive transurethral resection, 2 cycles of M-VAC chemotherapy, and radiotherapy (45 grays [Gy] on pelvic volume) with concurrent cisplatin (20 mg/m2 on Days 1-5. Response was determined by cystoscopic evaluation. If there was a complete response, radiotherapy continued until a total dose of 65 Gy; if there was not a complete response, cystectomy was performed. RESULTS: Tumor response after a dose of 45 Gy included 11 complete responses (55%), 5 partial responses (25%), and 4 nonresponses (20%). Overall survival was 75%, 34%, and 27% in the 2nd, 3rd, and 5th years of follow-up, respectively. Cause specific survival was 79%, 54%, and 38%, respectively. Survival for patients with complete response was 100%, 60%, and 48%, respectively. Severe toxicity was uncommon, with the most frequent toxicities being leukopenia and cystitis. No treatment-related death occurred with either treatment protocol. CONCLUSIONS: The age of the individual must not become a strict exclusion criterion for the radical treatment of old patients with invasive bladder carcinoma. PMID- 9210718 TI - Influence of nativity on cancer mortality among black New Yorkers. AB - BACKGROUND: Cancer is the second leading cause of death in U. S., and blacks have higher cancer death rates than whites. The authors conducted an analysis to determine the influence of birthplace on cancer mortality among blacks in New York City. METHODS: Death records for New York City from 1988 through 1992 were linked to the 1990 U. S. Census data. Age-adjusted cancer death rates by race and birthplace were computed. The experience of black residents born in the South and Northeast of the U. S. and in Caribbean countries were compared with that of New York City whites. RESULTS: The cancer mortality rate of blacks exceeded that of whites for males (512.6 vs. 385.6 per 100,000 per year), but was similar for females (270.8 vs. 270.6). However, cancer death rates of Southern-born black males (615.7) were substantially higher than those of black males born in the Northeast (419.1) or the Caribbean (352.4). Carcinomas of the lung, prostate, breast, and colon/rectum accounted for >50% of all cancer deaths. Lung carcinoma mortality varied greatly by birthplace, with Caribbean-born blacks (63.5 and 19.2 for males and females, respectively) having approximately one-third the death rates of Southern-born blacks (187.8 and 52.5 for males and females, respectively), and <50% that of New York City whites (108.7 and 53.2 for males and females, respectively). These differences were present in each age category, but were most pronounced among those age 45-64 years. In striking contrast, death rates from prostate carcinoma were highest in Caribbean-born black men, and this was especially apparent in persons age > or = 65 years. CONCLUSIONS: The generally higher cancer mortality of blacks compared with whites masks even greater intraracial heterogeneity revealed through stratification by birthplace. In general, Caribbean-born blacks are at lower risk of cancer mortality than other blacks, and whites, but their advantage does not hold for prostate carcinoma, for which Caribbean-born men had the highest mortality rate. PMID- 9210717 TI - Association between Epstein-Barr virus and Burkitt's lymphoma in Taiwan. AB - BACKGROUND: The association between Epstein-Barr virus (EBV) and Burkitt's lymphoma (BL) in Taiwan is not clear. In this study, the authors attempted to determine the frequency of the occurrence of EBV infection in patients with BL in Taiwan. METHODS: A retrospective study was performed using a nonisotopic in situ hybridization technique to detect the presence of EBV-encoded small RNAs (EBERs) in paraffin embedded BL tissues. Tissues of other types of B-cell non-Hodgkin's lymphoma (NHL) were used as controls. Immunohistochemical staining was performed to examine the presence of an EBV-encoded protein, latent membrane protein (LMP), and p53 in specimens. RESULTS: EBERs were detectable in 10 of 18 BL specimens. It was present in the cervical lymph nodes (LNs) in six of the seven cases of cervical tumors, in the maxillary region in one case, in one of two cases of axillary LNs, and in the abdominal tumors in two of the seven cases of intraabdominal disease. EBER positive cells were diffusely present in all tumors except in one abdominal BL, in which only a few EBER positive cells were scattered in a small part of the tumor. EBER positive cells were not detected in the case with BL in an inguinal LN and in the seven cases with intraabdominal tumors. Immunohistochemical studies showed that LMP and p53 were expressed in 3 and 4 of the 18 cases, respectively. In another 20 NHLs in peripheral LNs, EBERs were detectable in only 1 case of diffuse large cell histology with numerous reactive T cells in which only large tumor cells expressed EBERs and LMP. EBERs were not detected in any of the ten cases of extranodal NHL. CONCLUSIONS: In Taiwan, EBV is frequently associated with BL occurring outside the abdomen but rarely with intraabdominal BL. The overall association between EBV and BL in Taiwan is intermediate compared with other regions of the world. These results support the theory that the frequency of EBV associated with BL is influenced by the endemicity of EBV and/or the socioeconomic status of a country. PMID- 9210719 TI - Estimating the prevalence of cancer in the United States. AB - BACKGROUND: Few reports have estimated the prevalence of persons in the U.S. ever diagnosed with invasive cancer. METHODS: The Connecticut Tumor Registry was used to identify all Connecticut residents ever diagnosed (1935-1994) with invasive cancer who were known to be alive in 1994. Estimated prevalence rates for Connecticut were compared with those for 1982, and were applied to the total U.S. population for selected years. RESULTS: Some 95,361 persons ever diagnosed with invasive cancer(s) were confirmed as being alive at the end of 1994. The age standardized prevalence rate had increased by 40% in males and 13% in females since 1982, due in part to large increases for breast, prostate, and (in females) lung carcinoma. Using the data for Connecticut, an estimated 7.1 million Americans in 1995 had ever been diagnosed with invasive cancer; projected numbers were 7.7 million for 2000 and 13.2 million for 2030. CONCLUSIONS: The prevalence of persons ever diagnosed with invasive cancer could increase considerably in the coming decades, and numbers for elderly males could surpass those for elderly females by 2020. Although projections must be interpreted with caution, these data emphasize the need for primary prevention of cancer and for studies of cancer survivors. PMID- 9210720 TI - Diencephalic syndrome and disseminated juvenile pilocytic astrocytomas of the hypothalamic-optic chiasm region. AB - BACKGROUND: Diencephalic syndrome (DS) is a complex of signs and symptoms related to hypothalamic dysfunction; its main features are emaciation, despite a normal or slightly diminished caloric intake, and an alert appearance. DS has been almost exclusively described in association with space-occupying lesions of the hypothalamic-optic chiasm region, mainly juvenile pilocytic astrocytoma (JPA). A systematic diagnostic approach, including contrast-enhanced magnetic resonance imaging (MRI) of the child's head, is rapidly expanding our knowledge of this syndrome. METHODS: The MRI findings for three children affected by DS associated with biopsy-proven JPA, consecutively referred to the Pediatric Neuro-Oncology Program of the Department of Pediatrics at the University of Padua between September 1991 and January 1996, are presented in this article. The children were boys, ages 6, 7, and 18 months, respectively. RESULTS: In all three patients, the initial contrast-enhancing MRIs of the head showed evidence of tumor dissemination. This finding prompted a study of the spine, which in turn showed tumor deposits in all three subjects. Among the 43 patients younger than 16 years with low grade astroctyoma who consecutively entered the Neuro-Oncology Program during the study period, these 3 patients were the only ones who had disseminated tumors. CONCLUSIONS: In this study, the hypothesis was formulated that DS and disseminated hypothalamic-optic chiasm JPA tend to be more commonly associated than previously stated. This study suggests that the initial contrast-enhanced MRI of the head of a child affected by DS and hypothalamic JPA must be looked at carefully for evidence of tumor dissemination, and that the spine must also be examined if the findings are positive. PMID- 9210721 TI - Pleuropulmonary blastoma: a clinicopathologic study of 50 cases. AB - BACKGROUND: Pleuropulmonary blastoma (PPB) is a unique dysontogenetic neoplasm of childhood that appears as a pulmonary and/or pleural-based mass and is characterized histologically by a primitive, variably mixed blastematous and sarcomatous appearance. METHODS: Histologic material from all cases was reviewed and the tumors subclassified as type I (purely cystic), type II (cystic and solid), or type III (purely solid). Data regarding presenting symptoms, family history, operative findings, pathologic subtypes, therapeutic interventions, and outcome were correlated with survival by standard statistical methods. RESULTS: The series was comprised of 24 males and 26 females. Respiratory difficulty with or without fever was the most common clinical symptom reported. Cyst formation in the affected lung was identified radiographically in 19 children (38%) at or before the definitive pathologic diagnosis. The ages at presentation of the 7 type I, 24 type II, and 19 type III PPBs were significantly different: 10, 34, and 44 months, respectively (P < 0.001). Local recurrence developed in 1 of 7 type I PPBs (14%) and in 18 of 43 type II and III PPBs (46%); distant metastasis occurred in 13 patients, chiefly to the brain/spinal cord or bone, and was observed only in those with type II or type III PPB. Patients with pleural or mediastinal involvement fared significantly worse than those without such involvement. Five-year survival was 83% for type I and 42% for types II and III. Survival differences on the basis of pathologic subtype did not reach statistical significance. CONCLUSIONS: PPB is an aggressive, intrathoracic neoplasm of early childhood with an unfavorable outcome. Although survival differences among patients with different histologic subtypes of disease did not reach statistical significance, the apparently better outcome for patients with purely cystic type I tumors may be borne out in a large series. These observations support the premise that type I and III PPB are bridged morphologically by type II PPB with its combined cystic and solid features. The PPB should be regarded as the pulmonary dysontogenetic analogue to Wilms' tumor in the kidney, neuroblastoma in the adrenal gland, and hepatoblastoma in the liver. Molecular genetic investigations, especially in constitutional PPB, should be revealing. In view of the poor outcomes for patients with types II and III, new and aggressive therapies must be developed. PMID- 9210722 TI - The National Cancer Data Base report on the results of a large nonrandomized comparison of breast preservation and modified radical mastectomy. AB - BACKGROUND: Although the conclusions reached in the National Surgical Adjuvant Bowel and Breast Protocol B-06 trial and other clinical trials appear to remain intact, questions persist regarding the equivalency of breast preservation compared with modified radical mastectomy for patients with invasive carcinoma. Documentation and assessment of comparative survival rates in a large cohort of nonrandomized breast carcinoma patients was undertaken to understand better these outcome patterns. METHODS: Information gathered from the medical records of 96,030 women diagnosed with early stage carcinoma of the breast between 1985 and 1988 was reviewed to determine the age at diagnosis; tumor stage, grade, dimension; treatment; and disease status. RESULTS: Of these 96,030 Stage I and II (based on the American Joint Committee on Cancer staging system) patients, 8583 (8.9%) were treated with segmental mastectomy, axillary lymph node dissection, and radiotherapy without systemic treatment. Three thousand seven hundred and ninety-seven patients (4.0%) were treated with segmental mastectomy, axillary lymph node dissection, radiotherapy, and systemic therapy. Forty-four thousand two hundred and forty-nine patients (46.0%) were treated with modified radical mastectomy without systemic therapy, and 18,322 patients (19.1%) were treated with modified radical mastectomy with systemic therapy. Within each stage, reported survival was equal to or more favorable for patients managed with breast preservation compared with those treated with modified radical mastectomy. This comparability was observed in all subsets analyzed including those defined by age at diagnosis, histologic grade, and tumor dimension. CONCLUSIONS: These findings are consistent with the hypothesis that AJCC Stage I and II patients treated with breast preservation appear to have survival rates equivalent to those treated with modified radical mastectomy. PMID- 9210723 TI - Symptom survey: initial, critical step in a comprehensive community health study plan. PMID- 9210724 TI - Middle-ear disease in children exposed prenatally to polychlorinated biphenyls and polychlorinated dibenzofurans. AB - During 1978 and 1979, an episode of poisoning from ingestion of rice oil contaminated with polychlorinated biphenyls and polychlorinated dibenzofurans occurred in central Taiwan. We followed-up children who had been born between June 1978 and March 1985, as well as matched unexposed children. The mothers of exposed children had consumed contaminated oils before the children were born. In 1993, otolaryngologists examined the middle ear of each child with a pneumatic otoscope, and they measured the middle-ear pressure by tympanometry with a Rion RS20 impedance audiometer. The exposed children had a significantly higher prevalence of middle-ear diseases than their matched controls. The exposed children who had ear diseases had higher serum levels of 2,3,4,7,8-pentachloro- and 1,2,3,4,7,8-hexachloro-dibenzofurans than the children who did not have similar diseases. Therefore, in this study, children exposed to polychlorinated biphenyls and polychlorinated dibenzofurans had a higher incidence of middle-ear diseases than their controls. PMID- 9210725 TI - Fiber analysis in lungs of residents of a Japanese town with endemic pleural plaques. AB - The authors analyzed various types of fibers in lung-tissue samples, which were obtained from 50 cases (46 surgical resections and 4 autopsies) at Kumamoto Minami Hospital in Matsubase, where the occurrence of pleural plaques is endemic. Lung cancer necessitated surgical resection in 44 cases. Eleven of the 50 cases were residents of Matsubase; 15 resided in the region around the town, where the frequency of pleural plaques was slightly higher; and 24 cases lived in a region with normal plaque frequency. The number of anthophyllite fibers in the lungs of town residents was significantly higher than in residents of the region with normal plaque frequency. In 6 cases, the authors found accompanying pleural plaques, and the anthophyllite fiber count in the lungs in these cases was significantly higher than in cases without plaques. In addition, the anthophyllite fiber counts in 2 cases with severe plaques were significantly higher in 4 cases with only mild plaques. These results suggested that anthophyllite fiber might be responsible for the increased prevalence of pleural plaques in Matsubase. Even though the anthophyllite fibers were quite long (mean length = 25.1 microm), the width of most anthophyllite fibers were thick (mean diameter = 0.84 microm). Therefore, the aspect ratio of anthophyllite (mean = 38.7) was lower than that of amosite (mean = 81.8), which, in a previous report, was found predominantly in cases of pleural mesothelioma. Perhaps these differences in fiber size are related to the strength of the carcinogenicity to the pleura. PMID- 9210726 TI - Effects of occupational and nonoccupational factors on liver function tests in workers exposed to solvent mixtures. AB - A total of 368 workers from six paint-manufacturing factories participated in this study. The workers were classified according to type of exposure: direct, intermittent, and no exposure. The workers' liver-function tests were influenced greatly by gender, hepatitis B, alcohol consumption, and body mass index. Both the serum concentration and the odds of abnormality of total serum bile acids were elevated among the directly exposed group. The authors concluded that analysis of covariance should take into account occupational and nonoccupational factors on liver-function tests to avoid any errors. Total serum bile acids also indicated liver dysfunction from solvent exposure. PMID- 9210727 TI - A cluster of childhood leukemia near a nuclear reactor in northern Germany. AB - Between February 1990 and December 1995, professionals diagnosed six cases of childhood leukemia among residents of the small rural community of Elbmarsch in northern Germany. Five of these cases were diagnosed in only a 16-mo period between February 1990 and May 1991. All cases lived in close proximity (i.e., 500 4,500 m) to Germany's largest capacity nuclear boiling-water reactor. We calculated standardized incidence ratios and exact 95% confidence intervals for a 5-km-radius circular area around the plant. The standardized incidence ratio for the time period 1990-1995 was 460 (95% confidence interval: 210, 1,030). The analysis was restricted further to the years 1990 and 1991, and the standardized incidence ratio increased to 1,180 (95% confidence interval: 490, 2,830). Presently, this cluster of childhood leukemia cases cannot be explained in terms of established and putative risk factors--including radiation from medical sources--for childhood leukemia. PMID- 9210728 TI - Airborne (1-->3)-beta-D-glucan and airway disease in a day-care center before and after renovation. AB - Changes in symptoms and airway responsiveness among persons who worked in a day care center that had microbial growth problems were assessed before and after renovation. Before and after the building renovation, the investigators used the Limulus assay with (1-->3)-beta-D-glucan-specific lysate to measure airborne levels of (1-->3)-beta-D-glucan, a cell-wall component of molds. Airway responsiveness and subjective symptoms were measured among 14 female employees with a methacholine test and a standardized questionnaire. After the renovation, (1-->3)-beta-D-glucan-glucan levels decreased from 11.4 to 1.4 ng/m3. The number of persons who had increased airway responsiveness decreased after the renovation. Two employees developed a classical allergy to cat and pollen during the observation period. Although the study included only a few subjects and was based on only one day-care center, the data suggest that (1-->3)-beta-D-glucan may be related to airways inflammation caused by indoor air pollution. PMID- 9210729 TI - Blood lead level, by year and season, among poor pregnant women. AB - The authors conducted a longitudinal study of poor pregnant women and their infants to examine the determinants of maternal and infant lead levels. To accurately depict these determinants, one must account for secular and seasonal variations in these levels. The women's lead levels declined over the 5-y period of study by approximately 20%/y, depending on when in the course of pregnancy measurements were made. After correction for secular trend, we found a periodic effect that differed from that typically seen in children (i.e., peak occurs in summer). In this study, lead levels in these women peaked during December-March. If the effects of lead are greatest in the youngest conceptus, early pregnancies that occur in the December-March period pose the largest prenatal risk. PMID- 9210730 TI - Industrial noise exposure, noise annoyance, and serum lipid levels in blue-collar workers--the CORDIS Study. AB - Chronic noise exposure may constitute a risk factor for cardiovascular disease, but the exact mechanism is unclear. The authors studied the association between industrial noise exposure, noise annoyance, and serum lipid/lipoprotein levels in male (n = 1,455) and female (n = 624) blue-collar workers. The authors found that young men (i.e., < or = 44 y of age) exposed to high noise levels (> or = 80 dB[A]) had higher total levels of cholesterol (p = .023) and triglycerides (p = .001), as well as a higher cholesterol ratio (p = .038), than men exposed to low noise levels, even after controlling for confounding variables. In women or in older (> 45 y) men, noise did not affect serum lipid/lipoprotein levels. The authors found no interaction between noise exposure level and noise annoyance (except for high-density lipoprotein in women). However, noise annoyance covaried independently with total cholesterol (p = .022) and high-density lipoprotein (p = .0039) levels in young men and with total cholesterol (p = .035), triglyceride (p = .035), and high-density lipoprotein levels in women (under high noise exposure conditions)(p = .048) levels in women. Noise annoyance and noise exposure levels had an additive effect on cholesterol levels. Young men who scored high on both variables had a 15-mg/dl higher mean cholesterol level (95 % confidence interval [CI] = 7.2, 22.8; p = .0003) than those who scored low on both variables; in women, the corresponding difference was 23 mg/dl (95% CI = 1.5, 42.9; p = .019). The authors concluded that the examination of serum lipid/lipoprotein levels may be useful in studies of the health effects of noise, and particular attention should be paid to noise-annoyed individuals. PMID- 9210731 TI - Proportional mortality of dichloro-diphenyl-trichloroethane (DDT) workers: a preliminary report. AB - The authors conducted a proportional mortality study of 1,043 deaths that occurred between 1956 and 1992 among men who used mainly dichloro-diphenyl trichloroethane (DDT) in an anti-malarial campaign in Sardinia, Italy, during the late 1940s. For each cause of interest, investigators compared observed deaths with expected deaths. The estimated DDT exposure ranged from 170 to 600 mg/m3 in indoor operations and from 24 to 86 mg/m3 in outdoor operations. Workers directly exposed to DDT had a significant increase in risk for liver and biliary tract cancers (PMR = 228; 95% confidence interval = 143, 345) and multiple myeloma (PMR = 341; 95% confidence interval = 110, 795). However, the PMR for liver and biliary tract cancers was also elevated among workers who did not have direct occupational contact with DDT, and the authors observed no increase in either PMR, by number of days in exposed jobs. Perhaps DDT did not increase the risk or perhaps occupational exposure, although quite high, did not further increase the risk, compared with the heavy baseline exposure of the entire Sardinian population, (i.e., mainly through diet and drinking water). Expansion of the cohort to include all exposed workers, and collection of information to improve exposure assessment are needed to clarify these findings. PMID- 9210732 TI - Mouse lung immune response after acute exposure to flour dust. AB - To approach the physiopathology of the mucosal immune response in flour-mill and bakery workers, the authors developed a mouse model, which allowed them to study immune responses induced in the deep lung by acute inhalation of flour dust. In situ quantification of T lymphocytes (Thy1-2) and T-cell subsets (CD4 and CD8), macrophages, B lymphocytes, and immunoglobulin A plasma cells in the lungs of all animals revealed significant modifications of these cell compartments as early as 3 d after exposure; within 10 d of exposure, levels returned to baseline. The data suggest that the lung could be a sensitive target for inhaled xenobiotics, which might generate rapid immune local modifications that result in the maintenance of homeostasis. PMID- 9210733 TI - Dioxin-like compounds in fishing people from the Lower North Shore of the St. Lawrence River, Quebec, Canada. AB - In this study, investigators assessed exposure to dioxin-like compounds in a fishing population that inhabits small coastal communities along the Lower North Shore of the St. Lawrence River, Quebec. This population relies heavily on wildlife foods for sustenance. Investigators analyzed chemically the most popular marine foods (i.e., fish, crustaceans, sea mammals, and sea-bird eggs), and they also obtained 25 human plasma samples from individuals in two villages along the river. The mean level of total polychlorinated biphenyls in this population was approximately twice that found in the entire fishing cohort. Plasma levels of dioxin-like compounds, expressed as tetrachlorodibenzodioxin toxic equivalents, were approximately eight times higher than levels in urban residents. Most of the increase in tetrachlorodibenzodioxin toxic equivalents in the selected fish eaters resulted primarily from an elevation in polychlorinated biphenyls. Concentrations of dioxin-like compounds from the Lower North Shore were low in fish and seals, but concentrations were elevated in the eggs of sea birds. Given that there was also a significant statistical correlation in the entire population between human plasma levels and consumption of birds' eggs-and not other traditional foods-much of the increased human dose appeared to originate from this one food source. Because there appear to be increased, but uncertain, health risks from this elevated body burden, investigators advised the residents of the area to avoid consumption of wild birds' eggs (i.e., a food source of minor nutritional importance). PMID- 9210734 TI - Increase in child behavior problems resulting from maternal smoking during pregnancy. AB - In this article, the authors investigated the effect of maternal smoking during pregnancy on behavioral problems, which were not mediated by lower birth weight, in offspring at 3 y of age. The authors used the Child Behavior Checklist for ages 2-3 y (CBCL/2-3; Achenbach, Edelbrock and Howell) to assess behavioral problems in 1,377 2- to 3-y-old healthy twin pairs. Soon after the birth of twins, the authors collected pre- and perinatal information, including smoking habits of the mother during pregnancy. The question "Did you smoke during pregnancy?" could be answered by choosing one of three possible options: (1) never, (2) sometimes, or (3) regularly. The authors analyzed the effect of maternal smoking on the Child Behavioral Checklist total score and on several subscale scores for first- and second-born twins separately, and they adjusted for the possible confounding effects of birth weight, socioeconomic status, maternal age, and having been breast- or bottle-fed. There was a significant effect of maternal smoking on so-called externalizing behavior problems (oppositional, aggressive, overactive), but not on internalizing behavior problems (withdrawn, depressed, anxious), in both first- and second-born twins. The authors primarily attributed the enhanced externalizing problems to increased aggression. Although boys had higher externalizing (and aggression) scores than girls, the effect of maternal smoking was the same for boys and girls. The authors also discuss whether maternal smoking causes externalizing behavior problems. PMID- 9210735 TI - Brief communication: childhood leukemia in a residential small town near Barcelona. AB - Between March 1991 and May 1995, physicians diagnosed four cases of acute lymphoblastic leukemia, one case of Hodgkin's disease, and one case of aplastic anemia among children who resided in a small town near Barcelona that contained 4,237 inhabitants. The four cases of acute lymphoblastic leukemia represented a significant excess of observed cases (26.4/100,000 for children age 0-14 y [p < .005]). The authors conducted an epidemiological study of the population to explore the possible "local" role of agents hypothesized or known to be potentially associated with acute lymphoblastic leukemia, as well as with other hematopoietic diseases. The small town in which the cases lived is a residential area without known or suspected industrial exposures associated with leukemia. However, it is located in a county ("Maresme") that boasts having the most flower growing and agricultural undercover producing area in Catalonia; consequently, copious amounts of herbicides and pesticides are used. The small number of cases limited the testing of the hypothesis of a causal relationship between environmental pesticide exposure or a viral infection (the only factors common to the cases) and the excess of leukemia cases. Despite the weaknesses inherent in our study, the information gleaned from our research may be useful to researchers who define local health-related problems. PMID- 9210736 TI - Finding the "person" in personality disorders. PMID- 9210737 TI - Images in psychiatry. Rauwolfia serpentina: the first herbal antipsychotic. PMID- 9210738 TI - Divergences between clinical and research methods for assessing personality disorders: implications for research and the evolution of axis II. AB - OBJECTIVE: The purpose of this study was to examine the extent to which instruments for assessing axis II diverge from clinical diagnostic processes. METHOD: Subjects in the first study were 52 clinicians with experience in assessment and treatment of patients with personality disorders, who were surveyed about the methods they use in clinical practice to make diagnoses and other aspects of the diagnostic process. A second study replicated the major findings with a random national sample of 1,901 experienced psychiatrists and psychologists. RESULTS: Whereas current instruments rely primarily on direct questions derived from DSM-IV, clinicians of every theoretical persuasion found direct questions useful for assessing axis I disorders but only marginally so for axis II. They made axis II diagnoses, instead, by listening to patients describe interpersonal interactions and observing their behavior with the interviewer. In contrast to findings with current research instruments, most patients with personality disorders in clinical practice receive only one axis II diagnosis, and if they receive more than one, one is considered primary. Clinicians reported treating a substantial number of patients for enduring personality patterns that current axis II instruments do not assess, many of which meet neither axis I nor axis II criteria, notably problems with relatedness, work, self-esteem, and chronic subclinical depressive traits. CONCLUSIONS: Measurements of axis II were constructed by using a model derived from axis I instruments that diverges from clinical diagnostic procedures in a way that may be problematic for the assessment of personality disorders and the development of a more clinically and empirically sound taxonomy. PMID- 9210739 TI - Diagnostic criteria for complicated grief disorder. AB - OBJECTIVE: Some prolonged and turbulent grief reactions include symptoms that differ from the DSM-IV criteria for major depressive disorder. The authors investigated a new diagnosis that would include these symptoms. METHOD: They developed observer-based definitions of 30 symptoms noted clinically in previous longitudinal interviews of bereaved persons and then designed a plan to investigate whether any combination of these would serve as criteria for a possible new diagnosis of complicated grief disorder. Using a structured diagnostic interview, they assessed 70 subjects whose spouses had died. Latent class model analyses and signal detection procedures were used to calibrate the data against global clinical ratings and self-report measures of grief-specific distress. RESULTS: Complicated grief disorder was found to be characterized by a smaller set of the assessed symptoms. Subjects elected by an algorithm for these symptoms patterns did not significantly overlap with subjects who received a diagnosis of major depressive disorder. CONCLUSIONS: A new diagnosis of complicated grief disorder may be indicated. Its criteria would include the current experience (more than a year after a loss) of intense intrusive thoughts, pangs of severe emotion, distressing yearnings, feeling excessively alone and empty, excessively avoiding tasks reminiscent of the deceased, unusual sleep disturbances, and maladaptive levels of loss of interest in personal activities. PMID- 9210740 TI - Symptoms of obsessive-compulsive disorder. AB - OBJECTIVE: Obsessive-compulsive disorder encompasses a broad range of symptoms that represent multiple psychological domains, including perception, cognition, emotion, social relatedness, and diverse motor behaviors. The purpose of these analyses was to evaluate the correlational relationships of the symptoms of obsessive-compulsive disorder. METHOD: This study examined the 13 a priori categories used to group types of obsessions and compulsions in the Yale-Brown Obsessive Compulsive Scale symptom checklist in two independent groups of patients with obsessive-compulsive disorder (N = 208 and N = 98). A principal components factor analysis with varimax rotation was performed, followed by a series of other exploratory analyses. RESULTS: The two data sets yielded nearly identical results. Four factors--obsessions and checking, symmetry and ordering, cleanliness and washing, and boarding--emerged in each data set, in total accounting for more than 60% of the variance. CONCLUSIONS: Obsessive-compulsive disorder is a multidimensional and etiologically heterogeneous condition. The four symptom dimensions identified in this study are largely congruent with those identified in earlier reports. These factors may be of value in future genetic, neurobiological, and treatment response studies. PMID- 9210741 TI - Neuroanatomical correlates of externally and internally generated human emotion. AB - OBJECTIVE: Positron emission tomography was used to investigate the neural substrates of normal human emotional and their dependence on the types of emotional stimulus. METHOD: Twelve healthy female subjects underwent 12 measurements of regional brain activity following the intravenous bolus administration of [15O]H2O as they alternated between emotion-generating and control film and recall tasks. Automated image analysis techniques were used to characterize and compare the increases in regional brain activity associated with the emotional response to complex visual (film) and cognitive (recall) stimuli. RESULTS: Film- and recall-generated emotion were each associated with significantly increased activity in the vicinity of the medial prefrontal cortex and thalamus, suggesting that these regions participate in aspects of emotion that do not depend on the nature of the emotional stimulus. Film-generated emotion was associated with significantly greater increases in activity bilaterally in the occipitotemporparietal cortex, lateral cerebellum, hypothalamus, and a region that includes the anterior temporal cortex, amygdala, and hippocampal formation, suggesting that these regions participate in the emotional response to certain exteroceptive sensory stimuli. Recall-generated sadness was associated with significantly greater increases in activity in the vicinity of the anterior insular cortex, suggesting that this region participates in the emotional response to potentially distressing cognitive or interoceptive sensory stimuli. CONCLUSIONS: While this study should be considered preliminary, it identified brain regions that participate in externally and internally generated human emotion. PMID- 9210742 TI - Neuroanatomical correlates of happiness, sadness, and disgust. AB - OBJECTIVE: Happiness, sadness, and disgust are three emotions that differ in their valence (positive or negative) and associated action tendencies (approach or withdrawal). This study was designed to investigate the neuroanatomical correlates of these discrete emotions. METHOD: Twelve healthy female subjects were studied. Positron emission tomography and [15O]H2O were used to measure regional brain activity. There were 12 conditions per subject: happiness, sadness, and disgust and three control conditions, each induced by film and recall. Emotion and control tasks were alternated throughout. Condition order was pseudo-randomized and counterbalanced across subjects. Analyses focused on brain activity patterns for each emotion when combining film and recall data. RESULTS: Happiness, sadness, and disgust were each associated with increases in activity in the thalamus and medial prefrontal cortex (Brodmann's area 9). These three emotions were also associated with activation of anterior and posterior temporal structures, primarily when induced by film. Recalled sadness was associated with increased activation in the anterior insula. Happiness was distinguished from sadness by greater activity in the vicinity of ventral mesial frontal cortex. CONCLUSIONS: While this study should be considered preliminary, it identifies regions of the brain that participate in happiness, sadness, and disgust, regions that distinguish between positive and negative emotions, and regions that depend on both the elicitor and valence of emotion or their interaction. PMID- 9210743 TI - Reversed neurovegetative symptoms of depression: a community study of Ontario. AB - OBJECTIVE: Most research on depression with reversed neurovegetative features (hypersomnia, hyperphagia, and weight gain) has been based on site-specific clinic-based samples. The goal of this study was to delineate the epidemiology of reversed symptoms in a large community sample and to use other symptom patterns for comparison. METHOD: Interviewers assessed 8,116 subjects across Ontario, aged 15-64 years, by using the World Health Organization Composite International Diagnostic Interview. Individuals who met the DSM-III-R criteria for major depression, current or lifetime, were classified into four groups on the basis of lifetime neurovegetative symptoms: episodes of typical symptoms only, episodes of reversed symptoms only, neither type, or both types (fluctuating-symptom group). The groups were compared on demographic characteristics, comorbidity, disability, and health care utilization. RESULTS: Of the 653 individuals with lifetime major depression, 11.3% had episodes of reversed symptoms only, and another 5.8% were classified as fluctuating. Most of the differences among the four groups were due to the unique characteristics of the groups with neither type of episode or a fluctuating pattern; individuals who had experienced only reversed symptoms were remarkably similar to those who had had only typical symptoms. The fluctuating symptom group had high rates of comorbidity, substance abuse, and health care utilization. CONCLUSIONS: Several popular beliefs about depression with reversed features did not hold true for this community sample. Identifying individuals who fluctuate between reversed and typical episodes may be important in studies of major depression, in particular when reversed neurovegetative symptoms are a consideration. PMID- 9210744 TI - Vulnerability of Jews to affective disorders. AB - OBJECTIVE: Psychiatric literature over the past 100 years suggests that Jews are at higher risk for affective disorders than numbers of other religious groups. To examine these claims, the authors analyzed data from the National Institute of Mental Health Epidemiologic Catchment Area (ECA) study. In addition, the relationships among gender, alcoholism, and major depression were investigated. METHOD: The period prevalence and lifetime rates of DSM-III major depression among Jews, Catholics, Protestants, individuals in other religious groups, and individuals with no religious affiliation were examined in the Los Angeles and New Haven, Conn., ECA data. Logistic regression with covariates for site, gender, marital status, and socioeconomic status was used to estimate odds ratios and 95% confidence intervals. The calculated rates, based on the combined data from ECA study waves 1 and 2 for the white population, were weighted according to the 1980 U.S. population census. Female-to-male rate ratios and rates of alcohol abuse/dependence were also obtained. RESULTS: While no differences were found among females, Jewish males had significantly higher rates of major depression than Catholics, Protestants, and all non-Jews combined. Jews had a 1:1 female-to male ratio for major depression, in contrast to the other religious groups, which approached the universal 2:1 ratio. Rates of alcohol abuse/dependence were inversely related to rates of major depression. CONCLUSIONS: The results support only in part the earlier reports that Jews have higher rates of depression. The equal gender distribution of major depression among Jews may be associated with the lower rate of alcoholism among Jewish males. PMID- 9210745 TI - Comparison of induced and independent major depressive disorders in 2,945 alcoholics. AB - OBJECTIVE: Depressive episodes among alcohol-dependent men and women are heterogeneous in causation and clinical course. This study tested three hypotheses regarding the rates and clinical characteristics of two potential subtypes of these affective states: those that appear to be substance-induced mood disorders and those that are independent major depressive episodes. METHOD: Semistructured, detailed interviews were administered to 2,945 alcohol-dependent subjects as part of the Collaborative Study on the Genetics of Alcoholism. With the use of a time line method for determining the type of mood disorder among probands, relatives, and comparison subjects, individuals with histories of the two types of mood disorders were compared. RESULTS: Major depressive episodes with an onset before the development of alcohol dependence or during a subsequent long abstinence period (i.e., independent depressions) were observed in 15.2% of the alcoholics, while 26.4% reported at least one substance-induced depressive episode. According to a logistic regression analysis, the subjects with independent (as compared to substance-induced) major depressive episodes were more likely to be married, Caucasian, and female, to have had experience with fewer drugs and less treatment for alcoholism, to have attempted suicide, and, on the basis of personal interviews with family members, to have a close relative with a major mood disorder. CONCLUSIONS: These results support the contention that it is possible to differentiate between what appear to be substance-induced and independent depressive episodes in alcoholics. Such differentiation might be important for establishing prognosis and optimal treatment. PMID- 9210747 TI - Fluoxetine and norfluoxetine plasma concentrations in major depression: a multicenter study. AB - OBJECTIVE: Prior studies examining the relationship between fluoxetine plasma concentrations and response in major depression have either found no relationship between plasma concentration and response or suggested a curvilinear relationship with a therapeutic window. To elucidate this relationship, plasma concentrations of fluoxetine, norfluoxetine, fluoxetine plus norfluoxetine, and fluoxetine/norfluoxetine ratio were compared to therapeutic response. METHOD: A total of 839 patients (577 women, 262 men; mean age = 40 [SD = 11] with a DSM-III R diagnosis of major affective disorder who were in the course of either depression or bipolar disorder not otherwise specified and had a minimum baseline score of 16 on the 17-item Hamilton Depression Rating Scale were initially treated. Response was defined as follows: 1) nonresponders had less that a 50% or more reduction from baseline Hamilton depression score, 2) nonremitting responders had a 50% or more reduction from baseline Hamilton depression score but a final score higher than 7, and 3) remitters had a final Hamilton Depression score of 7 or lower. Plasma fluoxetine and norfluoxetine concentrations were measured after 8 weeks of fixed-dose treatment of 20 mg/day. RESULTS: Plasma concentration data were available from 615 patients. Plasma concentration were similar in responders, both remitting and nonremitting (N = 411), and nonresponders (N = 204) for fluoxetine concentrations, for norfluoxetine concentrations, as well as for the sum of fluoxetine and norfluoxetine and for the ratio of fluoxetine to norfluoxetine. No apparent relationship was observed between plasma drug concentration and clinical response. CONCLUSION: Plasma concentrations of fluoxetine and norfluoxetine do not appear to be related to clinical outcome and should not be used to make treatment decisions. PMID- 9210746 TI - Which elderly patients with remitted depression remain well with continued interpersonal psychotherapy after discontinuation of antidepressant medication? AB - OBJECTIVE: This study was conducted to identify which elderly patients with remitted recurrent major depression remain well with maintenance interpersonal psychotherapy after discontinuation of active antidepressant medication (nortriptyline). METHOD: The authors examined outcomes of maintenance therapy over 1 year for 47 elderly patients who were randomly assigned to monthly maintenance interpersonal psychotherapy with placebo (N = 19) or to placebo and a supportive medication clinic without interpersonal psychotherapy (N = 28). A Kaplan-Meier survival analysis was performed on the basis of treatment assignment and subjective sleep quality assessed by the Pittsburgh Sleep Quality Index, on which good subjective sleep quality is indicated by a score of 5 or lower. RESULTS: Nine (90%) of 10 patients reporting good subjective sleep quality (by 1 month into continuation treatment) remained well for at least 1 year when treated with monthly maintenance interpersonal psychotherapy, versus five (31%) of 16 patients with good sleep quality assigned to a medication clinic, three (33%) of nine patients with impaired sleep quality treated with maintenance interpersonal psychotherapy, and two (17%) of 12 patients with impaired sleep quality assigned to a medication clinic. CONCLUSIONS: Recovery of good subjective sleep quality by early continuation treatment is useful in identifying which remitted elderly depressed patients will remain well with monthly maintenance interpersonal psychotherapy, following discontinuation of antidepressant medication, and which patients may be more vulnerable to recurrence of major depressive episodes in the absence of antidepressant medication. PMID- 9210748 TI - Brainstem evoked response abnormalities in late-life depression with vascular disease. AB - OBJECTIVE: The purpose of this study was to examine whether the brainstem evoked responses of geriatric depressed patients with vascular disease show greater changes in wave V latency after increased stimulation than do responses of geriatric depressed patients without vascular disease and elderly comparison subjects with and without vascular disease. METHOD: Geriatric patients with unipolar depression (N = 53) were recruited from a university psychiatric hospital. Elderly comparison subjects (N = 23) were recruited through advertisements. All subjects were assessed for depressive symptoms, cognitive performance, overall medical burden, vascular disease, and disability. Brainstem evoked response was elicited at stimulation rates of 11.4 and 80.0 clicks/sec. RESULTS: The interaction between depression and vascular disease had a significant effect on change in wave V latency. This effect was synergistic, more than an additive effect. Post hoc comparisons showed that the depressed patients with vascular disease had greater changes in wave V latency that did the depressed patients without vascular disease, comparison subjects with vascular disease, and comparison subjects without vascular disease. Linear discriminant function analysis showed that 82% of the subjects with abnormal changes in wave V latency (sensitivity: 75%, specificity: 81%) could be identified on the basis of ratings for depression and vascular disease. CONCLUSIONS: Demyelination afflicting the pons and mesencephalon may explain the greater change in wave V latency for the brainstem evoked response in depressed patients with vascular disease. Further studies combining brainstem evoked response with brain imaging may determine whether depression develops only after vascular disease leads to demyelination. PMID- 9210749 TI - High intracellular calcium concentrations in transformed lymphoblasts from subjects with bipolar I disorder. AB - OBJECTIVE: Higher basal concentrations of intracellular calcium Ca2+ in platelets and lymphocytes from subjects with bipolar affective disorder than in unipolar depressed and healthy subjects implicate abnormal intracellular Ca2+ signaling in bipolar disorder. The purpose of this study was to clarify whether these intracellular Ca2+ abnormalities are trait related. METHOD: Basal Ca2+ concentration was measured by using ratiometric fluorescence assay with fura-2 for T lymphocytes and Epstein-Barr-virus-immortalized B lymphoblasts from physically healthy subjects with DSM-IV diagnoses of bipolar mood disorder (bipolar I, N = 28; bipolar II, N = 11) or major depressive disorder (N = 14), mixed psychiatric patients with non-mood disorders (N = 14), and health subjects (N = 20). Phytohemagglutinin-stimulated (10 micrograms/ml) intracellular Ca2+ levels were also determined in T lymphocytes. RESULTS: The basal Ca2+ concentration was significantly higher in the B lymphoblasts from patients with bipolar I disorder, but not bipolar II disorder or major depression, than in healthy subjects or psychiatric patients with nonmood disorders. There was a significant interaction between gender and diagnosis in the effect on basal Ca2+ levels in T lymphocytes. Contrasts of diagnoses within gender revealed significantly higher basal Ca2+ concentrations in T lymphocytes in male bipolar I patients, but not mean with bipolar II disorder or major depression, than in healthy male comparison subjects. Phytohemagglutinin-stimulated Ca2+ concentrations did not differ among groups, but the percent differences from basal Ca2+ levels were lower in bipolar I and depressed patients than in healthy subjects. CONCLUSIONS: These findings support the occurrence of abnormal calcium homeostasis in bipolar disorder and suggest that trait-dependent factors account, at least partly, for the higher basal lymphocyte Ca2+ concentration in bipolar I subjects. PMID- 9210750 TI - The firewater myth and response to alcohol in Mission Indians. AB - OBJECTIVE: This study aimed to assess empirically the intensity of reaction to alcohol in a group of Native Americans. METHOD: Forty healthy, nonalcoholic Mission Indian men between the ages of 18 and 25 years were tested before and after ingestion of placebo and 0.75 ml/kg of alcohol. Subjective (self-report of feelings) and objective (blood pressure, pulse rate, and plasma cortisol level) measures of intoxication were taken before ingestion of alcohol and placebo and at 15, 30, 60, 90, and 120 minutes after ingestion. Overall effects of alcohol were evaluated, and the responses of subjects with less than 50% Native American heritage (N = 19) were compared with the responses of subjects with at least 50% Native American heritage (N = 21). RESULTS: Alcohol did not produce any significant effects on any of the objective measures of intoxication; however, the subjects reported significant subjective effects of alcohol. Subjects with at least 50% Native American heritage reported less intense effects of alcohol than did those with less than 50% Native American heritage, despite equivalent blood alcohol concentrations. CONCLUSIONS: These results contradict the "firewater myth"--the theory that Native Americans are more sensitive to the effects of alcohol. Rather, the data indicate that Mission Indian men generally may be less sensitive to alcohol's effects, a physiological characteristic that has been shown to be associated with a greater risk for alcoholism in Caucasian populations. In addition, individuals with a greater percentage of Native American heritage may be less sensitive to the subjective effects of alcohol than individuals with a smaller percentage of Native American heritage. PMID- 9210751 TI - Somatization in cross-cultural perspective: a World Health Organization study in primary care. AB - OBJECTIVE: The purpose of this study was to examine the phenomenon of somatization in different cultures by determining its frequency and correlates in primary care settings in 14 countries. METHOD: Consecutive primary care patients (N = 25,916) were screened with the 12-item General Health questionnaire, and a stratified sample (N = 5,438) was interviewed with the Composite International Diagnostic Interview. Interviewed patients were also assessed for physical disease burden, self-rated overall health, physician-rated physical health status, number of disability days, and interviewer-rated occupational role functioning. The authors determined center-specific associations with the use of logistic regression analyses in which confounding variables were controlled. RESULTS: ICD-10 defined somatization disorder was relatively uncommon in most primary care settings. A less restrictively defined form was more common. Symptom rates were much higher in South American sites. There was a modest association with low education. Otherwise, frequency of unexplained somatic symptoms did not clearly vary according to geography or level of economic development. Somatizing patients were at elevated risk for self-reported disease burden, negative perception of their health, and comorbid depression and generalized anxiety disorder. Somatization was also commonly associated with disability. Cultures did not differ markedly in the pattern of these associated features. CONCLUSION: Somatization is a common problem in primary care across cultures and is associated with significant problems and disability. PMID- 9210752 TI - Refugees' time perspective and mental health. AB - OBJECTIVE: The authors' goal was to investigate factors protective of the mental health of refugees, with a particular focus on time splitting and suppression of the past. METHOD: Structured interviews covering premigration and postmigration stresses, personal and social resources, and mental health were given to 1,348 Southeast Asian refugees resettled in Vancouver, British Columbia, and to a comparison sample of 319 residents of Vancouver. Both groups of subjects also performed a task designed to measure orientation toward past, present, and future. RESULTS: Compared with resident Canadians, refugees were more likely to exhibit an atomistic time perspective in which past, present, and future are split. Temporal atomism and avoidance of nostalgia were associated with a lower risk of depression than were other time perspectives. CONCLUSIONS: Under conditions of extreme adversity, time splitting and suppression of the past may be adaptive strategies, mitigating the risk of depression. PMID- 9210753 TI - Traumatic grief: a case of loss-induced trauma. PMID- 9210754 TI - Cytoarchitecture of the entorhinal cortex in schizophrenia. AB - OBJECTIVE: The purpose of this study was to determine whether schizophrenia is associated with abnormalities in neuronal migration in the entorhinal cortex. METHOD: Nissl-stained sections through three cytoarchitectonic subdivisions of the entorhinal cortex were examined in postmortem brain specimens from 10 schizophrenic subjects and 10 matched normal comparison subjects. RESULTS: No qualitative differences in cytoarchitecture were observed between the schizophrenic and comparison subjects. CONCLUSIONS: These findings do not replicate previous reports of cytoarchitectural disturbances in the entorhinal cortex of schizophrenic subjects and thus fail to support the hypothesis of abnormal neuronal migration in schizophrenia. PMID- 9210755 TI - Schizophrenia and the parvalbumin-containing class of cortical local circuit neurons. AB - OBJECTIVE: The purpose of this study was to test the hypothesis that abnormalities in the parvalbumin-containing subclass of local circuit neurons contribute to altered gamma-aminobutyric acid (GABA) neurotransmission in the prefrontal cortex of schizophrenic subjects. METHOD: Profile counts and somal size measures were made of parvalbumin-immunoreactive neurons in areas 9, 46, and 17 from 15 matched pairs of schizophrenic and normal comparison subjects. RESULTS: No differences in relative density, laminar distribution, or somal size of labeled neurons were found in any region. CONCLUSIONS: These findings suggest that altered GABA neurotransmission in schizophrenia is due to either abnormalities in other sub-populations of prefrontal cortical GABA neurons or abnormalities in the parvalbumin-containing subclass that could not be detected in the present study. PMID- 9210756 TI - Olfactory identification deficits in schizophrenia: correlation with duration of illness. AB - OBJECTIVE: The authors examined the relationship between deficits in olfactory identification and duration of illness in young and elderly patients with schizophrenia. METHOD: Olfactory identification performance of 38 patients with schizophrenia and 40 normal subjects was compared by using the University of Pennsylvania Smell Identification Test. RESULTS: The schizophrenic patients demonstrated olfactory deficits relative to the comparison group, and the elderly schizophrenic patients displayed a greater magnitude of olfactory deficit than the younger patients. Independent of normal aging effects and cognitive deficit, patients with schizophrenia showed a strong relationship between olfactory identification scores and duration of illness, which suggests that olfactory abilities decline progressively over the course of the disorder. CONCLUSIONS: In contrast to other neuropsychological measures that have been reported to be stable over the course of illness, olfactory identification abilities deteriorate steadily in patients with schizophrenia, even for those with relatively recent onset. PMID- 9210757 TI - CSF neurotensin concentrations and antipsychotic treatment in schizophrenia and schizoaffective disorder. AB - OBJECTIVE: The relationship between CSF neurotensin concentrations and measures of psychopathology in patients with schizophrenia or schizoaffective disorder was examined before and after treatment with antipsychotic drugs. METHOD: CSF neurotensin concentrations were measured in 42 drug-free patients with schizophrenia or schizoaffective disorder. For 18 of these patients, CSF neurotensin was measure again after 4 weeks of antipsychotic treatment. RESULTS: Significantly higher levels of pretreatment psychopathology were observed in the patients with the lowest CSF neurotensin concentrations. Furthermore, improvements in overall psychopathology and, particularly, negative symptoms were correlated with increases in CSF neurotensin concentrations during treatment. CONCLUSIONS: These findings provide further evidence for a role of neurotensin the pathophysiology of psychosis and in the mechanism of action of antipsychotic drugs. PMID- 9210758 TI - Direct assessment of functional status in older patients with schizophrenia. AB - OBJECTIVE: There has been a growing trend in medicine to evaluate the impact of illness on functional abilities. Such studies typically rely on the patient's or caregiver's report. The goal of this study was to assess directly the functional capacity of psychiatric patients, especially older ones. METHOD: The subjects were 55 outpatients with schizophrenia and 72 normal persons ranging in age from 45 to 86 years. The subjects were administered the Direct Assessment of Functional Status Scale, which assess behaviour during simulated daily activity tasks in the areas of time orientation, communication, transportation, finance, shopping, grooming, and eating. RESULTS: The patients with schizophrenia had significantly greater disability than the normal subjects according to total scale scores as well as the communication, transportation, finance, and shopping subscale scores. Global cognitive status was the best predictor of total scale score. CONCLUSIONS: The Direct Assessment of Functional Status Scale is a promising instrument for functional assessment in outpatients with schizophrenia. PMID- 9210759 TI - Increased health care utilization as a function of participation in trauma research. AB - OBJECTIVE: This study was designed to compare, in a primary care setting, the health care utilization of women who participated in a trauma research study with the health care utilization of women who did not. METHOD: Health care utilization in the 12 months before and the 12 months after participation in trauma research was determined for both participants (N = 116) and a group of control subjects (N = 100) matched for day of service. RESULTS: Pairwise t test results indicated that for the women who participated in the research, all measures of health care utilization significantly increased in the 12 months after the trauma study; for the control subjects, only the number of ongoing prescriptions significantly increased. Sign tests confirmed that a significantly greater number of research participants demonstrated a positive difference (increase in utilization) for all health care variables, whereas only ongoing prescriptions demonstrated a significant systematic increase among control subjects. CONCLUSIONS: The findings suggest that participation in trauma research may increase subsequent health care utilization. PMID- 9210760 TI - Interpersonal psychotherapy for depressed antepartum women: a pilot study. AB - OBJECTIVE: Antenatal depression, a substantial risk factor for postpartum depression, occurs in 10% of pregnant women, but no clinical treatment trials of antenatal depression exist. In an effort to establish treatment guidelines for depression during pregnancy, the author reports on a treatment program using interpersonal psychotherapy for antepartum depression. METHOD: A 16-week open pilot trial conducted with 13 pregnant women who met DSM-III-R criteria for major depression. RESULTS: The women's mean depression ratings decreased significantly from week 0 to week 16 of the treatment program. CONCLUSIONS: Interpersonal psychotherapy for antepartum depression appears to be an effective alternative to pharmacotherapy in pregnancy. This study served as a pilot for an ongoing controlled clinical treatment trial. PMID- 9210761 TI - Paroxetine for treatment of obsessive-compulsive disorder and comorbid stuttering. PMID- 9210762 TI - Hallucinogen-induced relief of obsessions and compulsions. PMID- 9210763 TI - Nefazodone and visual side effects. PMID- 9210764 TI - Inpatient treatment of combat veterans. PMID- 9210765 TI - Comorbidity between personality and dysthymic disorders: historical and conceptual issues. PMID- 9210766 TI - Academic versus clinical views on lithium use. PMID- 9210767 TI - Androgens and alcohol. PMID- 9210768 TI - Anorexia nervosa in eighteenth century. PMID- 9210769 TI - The healing power of pets: a look at animal-assisted therapy. PMID- 9210770 TI - The power to do harm. PMID- 9210771 TI - Comments on the nature of prayer research. PMID- 9210772 TI - Who will care for the caregivers? PMID- 9210773 TI - Humanizing medicine: the humanities program at Marin General Hospital. PMID- 9210774 TI - A communication process: a new paradigm applied to high-dilution effects on the living body. AB - Living beings communicate with their world nonverbally, whether on a somatic or a psychological level. This paradigm of signifiers or sense takes place in the framework of the logic of analogy. The signifier is the semantic object that materially designates information to be transmitted and dealt with; a homeopathic remedy is the mimetic representation of the disease. Differential levels of information organize the spread of signifiers; each level is the result of regulation and integration of the previous level. The living self is the never ending process whereby levels of information are synthesized in the face of the informing environment. Such representations meet one another in the communication between the patient and the physician-remedy system. The medical device must reinform the patient and make the patient's signs and symptoms move toward a higher level of integration. The dilution of the remedy permits us to receive and treat it as information about disease. Signs and symptoms can be recognized as an erroneous adaptation; the organism is engaged in a process of paradoxical negation. The action of the remedies consists of a dynamic analogy between pieces of information. The paradigm of signifiers offers a new possibility for the exploration of informative therapeutics. PMID- 9210775 TI - Utilization of Chinese medicine in Taiwan. AB - This article examines the use of Chinese medicine in Taiwan. Based on a national sample survey, the authors investigated how Chinese medicine is being used and factors that are associated with its use among the Taiwanese. Results of the study suggest that Chinese medicine use among this population depends on health conditions, and that having a regular source of care for Chinese medicine as well as a preference for Chinese medicine are two predictors for its use. Policies on the integration of Chinese medicine or other traditional medicine into the modern healthcare system are recommended. PMID- 9210776 TI - Job stress reduction therapies. AB - BACKGROUND: Job stress among healthcare workers has received more attention in recent years, perhaps because these professionals are prime candidates for high stress levels. METHOD: The immediate effects of brief massage therapy, music relaxation with visual imagery, muscle relaxation, and social support group sessions were assessed in 100 hospital employees at a major public hospital. DESIGN: The effects of the therapies were assessed using a within-subjects pre post test design and by comparisons across groups. RESULTS: Groups reported decreases in anxiety, depression, fatigue, and confusion, as well as increased vigor following the sessions. CONCLUSION: That the groups did not differ on these variables suggests that these particular therapies, when applied for short periods of time, are equally effective for reducing stress among hospital employees. PMID- 9210777 TI - Comparing Hatha yoga with dynamic group psychotherapy for enhancing methadone maintenance treatment: a randomized clinical trial. AB - BACKGROUND: As more methadone treatment programs are funded in an attempt to curb substance abuse and HIV infection among i.v. drug users, more cost effective treatment approaches are being sought. OBJECTIVES: To investigate whether clients in outpatient methadone maintenance treatment who practice weekly Hatha yoga in a group setting experience more favorable treatment outcomes than those who receive conventional group psychodynamic therapy. METHODS: After a 5-day assessment period, 61 patients were randomly assigned to methadone maintenance enhanced by traditional group psychotherapy (ie, conventional methadone treatment) or an alternative Hatha yoga therapy (ie, alternative methadone treatment). Patients were followed for 6 months and evaluated on a variety of psychological, sociological, and biological measures. The revised Symptom Check List provided the primary psychological measures; the Addiction Severity Index provided various indices of addictive behaviors. RESULTS: The evidence revealed that there were no meaningful differences between traditional psychodynamic group therapy and Hatha yoga presented in a group setting. Both treatments contributed to a treatment regimen that significantly reduced drug use and criminal activities. Psychopathology at admission was significantly related to program participation regardless of treatment group. DISCUSSION: In addition to examining the characteristics of patients who present for treatment, this study identifies unexpected staff issues that complicate the integration of alternative and traditional treatment strategies. CONCLUSION: Alternative methadone treatment is not more effective than conventional methadone treatment, as originally hypothesized. However, some patients may benefit more from alternative methadone treatment than conventional methadone treatment. Additional research is necessary to determine characteristics that identify patients who might benefit from alternative methadone treatment. PMID- 9210778 TI - Joan Halifax, PhD. Being with death and dying. Interview by Bonnie Horrigan. PMID- 9210779 TI - Cultural diversity, folk medicine, and alternative medicine. PMID- 9210780 TI - Metaphors in the teaching of holistic medicine. PMID- 9210781 TI - Ultrastructure of rat Purkinje neurons after chronic ethanol consumption and prolonged abstinence. AB - In the present study of nutrition control of Wistar white rats, the ultrastructure of cerebellar Purkinje cells was studied after chronic ethanol exposure and a subsequent period of prolonged abstinence: a qualitative investigation of the perikarya of Purkinje cells was performed in age-matched controls (group A) and rats alcohol-fed for 5 months and withdrawn from this diet for 3 months (group B). The results showed massive accumulation of small dense bodies as well as obvious deposition of lipofuscin in the Purkinje cells of group B. Furthermore, ring-shaped Golgi apparatus units, lamellar bodies and degenerative foci dispersed throughout the cytoplasm of the alcohol-treated animals referred to degeneration processes and neuronal cell death, the morphological characteristics and the aetiology of which are discussed. PMID- 9210782 TI - Anatomo-radiographic study of prenatal development of bovine fetal teeth. AB - The aim of this study was to improve the knowledge of the time of appearance of dental germs and their morphological development until birth in bovine fetuses. Skulls and isolated mandibles of 35 Simmenthal bovine fetuses, of both sexes and ages from 97 to 280 days old were examined. The radiographic examination was performed with high definition and mamofilms. The exposure values ranged from 36 kV-6 mAs to 55 kV-12 mAs according to skull dimensions. In this study, the first dental germ was observed at 97 days, identified as the third maxillar premolar tooth. Through the morphological study and accurate description of the lingual and vestibular aspect of the occlusive surface of the teeth, three roots for the third and fourth maxillar premolar teeth and two for the second maxillar premolar tooth were observed. Two roots for the second and third mandibular premolar teeth and three for the fourth mandibular premolar tooth were also observed. The germ of the first mandibular molar tooth was seen at 140 days and the first of the maxillar arch at 280 days. PMID- 9210783 TI - Light and electron microscopic examination of various doses of given vitamin A on the development of cornea in mice. Partly presented in ICEM 13-Paris. AB - Vitamin A (0.2 micrograms, 0.6 micrograms, 1.2 micrograms) was administered orally to the mice on days 8-11 of gestation. Fetuses were removed on day 17 of gestation. No external malformation of the fetuses was seen on the stereomicroscope investigation. Corneal degeneration was seen on the light and electron microscopic examination. As a result it was accepted that vitamin A taken during the critical periods of gestation affected the development of cornea. PMID- 9210784 TI - The effect of formalin on rumen surface area in red deer. AB - Previously published studies have related the surface area of rumen wall to diet. The validity of studies that utilize preserved material depends upon the predictability of any change in rumen dimension brought about by preservation. Changes in the surface are of different sections of the rumen wall of red deer (Cervus elaphus) were monitored after immersion in 10% formal saline solution at room temperature for 2, 7 and 14 days. There was a high degree of variability (0 39.5%) in wall-area reduction. Ignoring such changes when calculating the factor of increase in surface area (FISA), a composite of papillary surface area and density, can result in errors of up to 39.5%. This study questions the validity of using FISA calculations when formalin-preserved specimens are used in studies of rumen response to diet. Rumen papillary dimensions were not significantly changed by preservation in formalin. PMID- 9210785 TI - Application of computer-assisted morphometry to the analysis of prenatal development of human lung. AB - Morphometry is well-established in tumour pathology. To evaluate is potential usefulness for description of developmental processes, histological slides from paraffin-embedded specimens of 67 human fetal lungs were Feulgen-stained, and morphometric characteristics of nuclei of epithelial pulmonary cells were analysed with an automated image analysis system. The measured cytometric features comprised of integrated optical density (IOD), S-phase-related IOD fraction, IOD entropy and nuclear area. Histometric features of the specimens were based upon the minimum spanning tree (MST) and included distances between neighboring epithelial cells, between epithelial cells and neighboring lymphocytes, and assessment of MST entropy. Notably, certain parameters revealed a non-uniform level during prenatal development S-phase-related IOD fraction increased from 5% to 8% between 14 and 16 weeks of gestation, then declined to 6% until birth. The IOD entropy steadily increased during development, whereas the extent of nuclear area remained constant. In accordance with an increase of the S phase-related fraction the MST entropy displayed a singular peak between 14 and 16 weeks of gestation, which is probably associated with development of glandular structures in the lung. Correlation of expression of binding sites for markers, presumably involved in functional aspects of development, with such alterations, is shown for binding capacities of biotinylated fucoidan and the S-phase-related fraction. This may be helpful to infer immuno- or ligando histochemically defined tissue sites with potential physiological significance in morphometrically distinguished periods of development. PMID- 9210786 TI - Morphometric comparison between contralateral sciatic nerves in the male and female rabbit. AB - Some morphometrical parameters of the axons making up the contralateral sciatic nerves, both in the male and female rabbits were calculated and compared by means of a Zeiss Vidas image analyser (Ober Kochen, Germany). The results show that the fibres constituting the left nerve have a greater mean diameter but a lower mean density that those constitution the right nerve. This suggests that the diameter of the myelinated fibres and the density of both the myelinated and unmyelinated fibres do not vary from male to female. On the other hand, the G ratio and the diameter of the unmyelinated axons do, since the nerves on the right side (in both sexes) have higher morphometric values, on average, than the contralateral ones. PMID- 9210787 TI - Sertoli cell-spermatid tubulobulbar complexes in the ram. AB - The fine structure of the tubulobulbar complex (TBC) of the ram seminiferous epithelium was studied in immersion-fixed testicular samples obtained in autumn (sheep's mating period). The TBC was visible during the last two stages of the seminiferous epithelium cycle. It originated as a tubular invagination of the Sertoli cytoplasm harbouring a complementary evagination for the spermatid head. Later, it expanded into a bulbous structure, finally becoming detached from the spermatid and apparently phagocytosed by the Sertoli cell. The significance of this transient structure is discussed and compared with previous reports for other eutherian mammals. PMID- 9210788 TI - The influence of 2-propanol and acetone on oviposition rate and oviposition site preference for acetic acid and ethanol of Drosophila melanogaster. AB - Drosophila melanogaster strains, homozygous for the alcohol dehydrogenase alleles AdhF, AdhS, and Adhn4 respectively, were tested for oviposition site preference with a Multiple Choice Device consisting of 18 patches per choice disk. Equal numbers of patches with ethanol-, acetic acid-, and water-supplemented medium were offered simultaneously. Patches with acetic acid-supplemented medium were chosen predominantly as oviposition sites. Pretreatment of flies with increasing concentrations of 2-propanol to inhibit alcohol dehydrogenase (ADH) activity resulted not only in a decreasing choice of acetic acid patches, but also in the laying of a decreasing number of eggs. Adh-null mutant flies showed a similar change in behavior pattern after 2-propanol treatment. Therefore it was concluded that ADH activity is not involved primarily in oviposition site preference behavior. A complicating factor is acetone, the oxidation product of 2-propanol, which had an even larger impact on egg production. However, differences in ADH allozymes with respect to biochemical oxidation capacity of secondary alcohols will not necessarily lead to differences between the Adh genotypes in oviposition rates or apparent changes in preferences, due to additional biochemical differences in inhibition rates by acetone of the various allozymes and other enzyme systems. PMID- 9210789 TI - Inheritance of hypoxic exercise tolerance in mice. AB - All mammals tested, when exposed acutely to a degree of hypoxia above some threshold, exhibit a reduced capacity to perform work. Chronic hypoxic exposure is usually associated with some degree of acclimation resulting in partial recovery of the preexposure work capacity. The present study reports that, among mice, interindividual variability in recovery of ability to tolerate a standardized hypoxic exercise [t(et); time elapsed in treadmill exercise in hypoxia until 4-s failure to avoid a grid configured to deliver a mild aversive current (0.15 mA)], after 8 weeks' exposure to half-atmospheric pressure, is influenced predominantly by two unlinked genes of major effect. Two approaches were taken toward genetic characterization. In one, a maximum-likelihood procedure was applied to 11 models of genetic determinacy in the t(et) distributions of BALB/cBy (C) and C57BL/6By (B6) parental inbred strains, their F1 hybrid, and the backcross (BC) generations. Breeding tests of the resulting candidate "best-fit" major locus inheritance models involved repeated cycles of selecting, as the progenitor of a new BC generation, the male with the highest value of the test variable in the previous BC generation, and breeding him to C females. Mice from each of four distinct phenotypes appearing in BC3 were bred to C mice, producing distributions expected from two-locus segregation. The second approach was based upon CXB/By RI strain distribution pattern and derivative breeding tests to reveal phenotypic distributions consistent with two-locus inheritance of tet. Melding these results with a positional cloning strategy may permit relating a behavioral difference to specific heritable elements and identifying their products as the (partial) physiological substrata of the behavior. PMID- 9210790 TI - Antipredator behavior in paradise fish (Macropodus opercularis) larvae: the role of genetic factors and paternal influence. AB - The paradise fish, a small insectivore, coinhabits marshes of Southeast Asia with several predator fish species. Its ability to recognize and avoid harmful fish may depend upon both genetic factors and experience. Here we demonstrate genetic variability between the 20-day-old larvae of two inbred strains of paradise fish (P and S) in predator exploration and avoidance, using predator models. We show that, in comparison to S larvae, P larvae exhibited an elevated frequency of leaping and backing and an increased approach latency when faced with a predator model with eyespots. Analysis of a classical cross system between the two strains revealed significant departure from an additive-dominance genetic model and suggested the involvement of both epistatic effects of several genes and paternal effects. The effect of the paternal influence during the 5-day postspawning period was found to be strain dependent: later predator avoidance behaviors were influenced by the presence of the father in P larvae but not in S larvae. On the basis of these and previous results, we speculate that the 5 postspawning days may represent a developmentally sensitive period during which specific environmental stimulation, e.g., stimuli associated with the father, is critical for later development of appropriate antipredatory responses. We conclude that developmental aspects of antipredatory behavior in paradise fish are influenced by a complex interplay between genetic and environmental factors. PMID- 9210791 TI - Mapping quantitative trait loci for open-field behavior in mice. AB - By performing a whole genome screen in an F2 intercross of two strains of mice (A/J and C57BL/6J), which differ markedly in their behavioral response to a brightly lit open field (O-F), we have mapped several quantitative trait loci (QTL) for this complex behavioral phenotype. QTL on chromosomes 1 and 10 were identified that affect both initial ambulation in the O-F (initial "response to novelty" ambulation) (lod of 7.1 and 8.8, respectively) and vertical rearings (lod of 4.5 and 8.5, respectively). For habituated O-F behavior, QTL were identified on chromosomes 3 and 10 for ambulation (lod of 4.1 and 14.7, respectively) and on chromosomes 1, 10, and 19 for vertical rearings (lod of 5.8, 6.0, and 4.7, respectively). The QTL on chromosome 1 (near D1Mit116; 101 cM) was specific for initial O-F ambulation behavior, whereas the QTL on chromosome 10 (near D10Mit237; 74 cM) affected both initial and habituated rearing behavior. Additional suggestive QTL (lod, > 2.8) were mapped to chromosomes 1, 8, 11, 15, and 19. The QTL on chromosomes 1, 10, and 19 individually explain from 3.2 to 12.7%. Collectively, the multiple independent QTL explain from 16.3 to 24.1% of the F2 population's phenotypic variance, depending on the trait. These identified QTL should prove useful for dissecting the genetic and behavioral dimensions of O F behavior, fostering an understanding of individual differences. PMID- 9210792 TI - Genes controlling ion permeability in both motorneurons and muscle. AB - Genetic data suggest that unc-8 is a member of the epithelial sodium channel (ENaC) gene family (shreffler et al., 1995). Consistent with this idea, cosmid R13A1, containing an ENaC homolog, can restore normal locomotion to unc-8 mutants after germline transformation. To identify other genes encoding proteins that regulate ENaC function, extragenic unc-8 suppressor and enhancer mutations were sought. This report describes two new unc-8 suppressor mutations, sup-42(lb88) X and sup-43(lb141) II, and an enhancer mutation, enu-2(lb140) III. sup-43(lb141) and enu-2(lb140), cause vacuoles within body wall muscle, similar in appearance to those of unc-105(n490) II mutants, consistent with their proposed role in ENaC function. Single and double mutant phenotypes observed in this and previous work suggest that sodium channels in different tissues utilize an overlapping set of gene products: at least six in motorneurons, unc-8, mec-6, and sup-40-43, and at least five in muscle, sup-43, unc-8, enu-2, unc-105, and mec-6. PMID- 9210793 TI - The effect of early experience on MHC-based mate preferences in two B10.W strains of mice (Mus domesticus). AB - Previous experiments have demonstrated that mice of some strains show mate preferences that are based on genes of the major histocompatibility complex (MHC), and rearing environment appears to influence these preferences. This experiment investigated if fostering affected MHC-based mate preferences in two additional strains of mice for which it was known that females exhibited MHC dissimilar preferences. Pups were exchanged between families of B10.GAA37 and B10.CHR51 mouse strains, which differed genetically from one another only at MHC loci. At sexual maturity foster mice were given a choice of two opposite-sex mice of either the foster-family or the foster-mouse MHC type. Preference was based on time spent with each stimulus mouse, the first ejaculation, or the first mount with a stimulus mouse. Although the results were not significant in general, females of the B10.GAA37 strain were mounted first significantly more often by non-foster-family males; first mounts predicted ejaculation preference. The results suggest that rearing environment did affect MHC-based preferences in females of this strain, although learning of self-MHC cues and use of non-MHC cues for mate choice may also occur. These results are compared to those of previous experiments. PMID- 9210794 TI - Nesting and fitness: lifetime reproductive success in house mice bidirectionally selected for thermoregulatory nest-building behavior. AB - To test the hypothesis that large, well-built, nests are an important component of fitness, we kept 12 mating pairs of two high-selected, two control, and two low-selected lines, selected for thermoregulatory nest-building behavior, at 22 and 4 degrees C with access to 10 g of cotton to build a nest, for a period of 180 days. Measurements included number of litters born per family, number of young per litter born and surviving up to 40 days of age, nest type built by the parents, and weight gain of the young from weaning (20 days of age) to 40 days of age. In all lines the production and survival of offspring was substantially decreased at 4 degrees C compared to 22 degrees C, but the high-selected lines produced more and better-quality offspring, surviving up to 40 days of age at both temperatures compared to the control and low-selected lines. This indicates that thermoregulatory nest-building behavior and evolutionary fitness are closely associated. PMID- 9210795 TI - Genetic and environmental contributions to the correlation between alcohol consumption and symptoms of anxiety and depression. Results from a bivariate analysis of Norwegian twin data. AB - Two thousand five hundred seventy pairs of Norwegian MZ and like-sexed and unlike sexed DZ twins aged 18-25 years completed questionnaires with information about symptoms of anxiety and depression and alcohol consumption. The aim of the study was to estimate sex-specific genetic and environmental effects unique to symptoms of anxiety/depression and to alcohol consumption and effects common to the two phenotypes. Five models fitted the data almost equally well. The heritability estimate from these models ranged from .23 to .57 for male alcohol consumption, from .39 to .59 for female alcohol consumption, from .25 to .48 for male anxiety/depression, and from .45 to .56 for female anxiety/depression. The phenotypic correlation between alcohol and anxiety/depression in males (r = .23) could be fully explained by common genetic effects. The correlation in females (r = .18) was caused by individual environmental factors together with either genetic effects or family environment. PMID- 9210797 TI - Diphtheria acquired during a cruise in the Baltic Sea: update. PMID- 9210798 TI - The incidence of gonorrhoea in England and Wales is rising. PMID- 9210796 TI - DNA by mail: an inexpensive and noninvasive method for collecting DNA samples from widely dispersed populations. AB - As specific genes are identified that are associated with behavior, it becomes increasingly important for behavioral geneticists to be able to incorporate these genes in their research. Rather than using blood, DNA can be extracted from cheek swabs, which makes it possible to obtain DNA inexpensively by mail from large, widely dispersed individuals. The purpose of this paper is to recommend this technique to the behavioral genetics community and to present results of our use of this technique to obtain DNA by mail for 114 2-year-olds and 116 adults. PMID- 9210799 TI - New horizons in facial nerve therapy. PMID- 9210800 TI - Attic cholesteatoma and tympanosclerosis. PMID- 9210801 TI - Laryngeal trauma and laryngeal pain. PMID- 9210802 TI - Inferior meatal antrostomy: is it still indicated? PMID- 9210803 TI - Etidronate for the the neurotologic symptoms of otosclerosis: preliminary study. AB - The efficacy of etidronate, a bisphosphonate, was assessed as a treatment for the inner ear symptoms of otosclerosis in a retrospective case review of 896 patients diagnosed with otosclerosis, with primary complaints of dizziness, hearing loss, tinnitus or Meniere's syndrome. The diagnosis of otosclerosis was based on small pixel computed tomography of the temporal bones. Of the 896 patients placed on an etidronate protocol, 545 were followed for more than six months and were analyzed. The symptomatic response to etidronate, as well as audiologic and computerized rotary chair results were used in the assessment. Patients who were previously on sodium fluoride were separately analyzed. In this preliminary study etidronate appeared to be an effective treatment for the neurotologic symptoms of otosclerosis. Prospective blinded efficacy studies of the bisphosphonates in the treatment of otosclerosis should be undertaken. PMID- 9210804 TI - Vocal process granuloma. AB - Vocal process granuloma or contact ulcer is uncommon disease in which there is chronic irritation and granulation tissue formation at the posterior third of the vocal folds. Thirteen patients (11 men and two women) with vocal process granuloma were enrolled in this study; cases of intubation granuloma were excluded. The most frequent complaints were throat irritation, frequent throat clearing and voice change. Forty-seven percent of patients had a recurrence two to four months after surgery. Computed tomography (CT) of the larynx in four patients showed arytenoid sclerosis on the involved side and disclosed moderate enhancement of the vocal fold granuloma after contrast injection in one. Three patients had hyperacidity and four had hyperfunctioning granulomas: two used their voices excessively and the other two had bilateral sulcus vocalis. To our knowledge this is the first report of sulcus vocalis with vocal process granuloma, and of enhanced vocal process granuloma. PMID- 9210805 TI - Compatibility of the CLARION cochlear implant with the connevans CRM-220. PMID- 9210806 TI - Bone conduction implants: Xomed Audiant bone conductor vs. BAHA. PMID- 9210807 TI - External unit for a semi-implantable middle ear hearing device. AB - A miniaturized, low-power external unit has been developed for the clinical trials of a semi-implantable middle ear electromagnetic hearing device (SIMEHD) which uses radio-frequency telemetry to couple sound signals to the internal unit. The external unit is based on a commercial hearing aid which provides proven audio amplification and compression. Its receiver is replaced by an application-specific integrated circuit (ASIC) which: 1) adjusts the direct current bias of the audio input according to its peak value; 2) converts the audio signal to a one-bit digital form using sigma-delta modulation; 3) modulates the sigma-delta output with a radio-frequency (RF) oscillator; and 4) drives the external RF coil and tuning capacitor using a field-effect transistor operated in class D. The external unit functions as expected and has been used to operate bench-top tests to the SIMEHD. Measured current consumption is 1.65-2.15 mA, which projects to a battery lifetime of about 15 days. Bandwidth is 6 kHz and harmonic distortion is about 2%. PMID- 9210808 TI - Rehabilitation in Franceschetti syndrome: an interdisciplinary approach using bone-anchored hearing aids. AB - The purpose of the study was to determine the effectiveness of a concept of combined interdisciplinary rehabilitation for children with mandibulofacial dysostosis, developed at the Center for Facial Malformations. It consists of binaural implantation of bone-anchored hearing aids and gradual distraction of the mandible. After audiological testing and mandibular distraction on a phantom head designed with data from a spiral CT, the surgery was done in three steps: implanting the fixtures for BAHA and the bone-lengthening device, removing the device after six weeks and completing the BAHA implantation two months later. The distraction procedure and orthodontic treatment were performed on an outpatient basis. The results (six patients, ages 6-19 years) were excellent: after implantation of the BAHA system speech perception increased from approximately 85% with the conventional BCHA to 95-100% with the BAHA. Quality of life was reported to be much better because of the general cosmetic improvement as well as the good acoustic orientation and sound quality with the new hearing devices. We conclude that the interdisciplinary approach provides favorable conditions for rehabilitation in cases of complex malformations of the head and neck. PMID- 9210809 TI - Late-onset life-threatening angioedema and upper airway obstruction caused by angiotensin-converting enzyme inhibitor: report of a case. AB - Angioedema is a rare but potentially lethal adverse effect when associated with upper airway obstruction. Sporadic cases of angioedema secondary to angiotensin converting enzyme inhibitors (ACEI) have been reported in the literature. The overall incidence is around 0.1% to 0.2%, and the time of onset is usually during the first week of ACEI therapy. Late-onset angioedema secondary to treatment with ACEIs is much more frequent than appreciated, and is largely unrecognized because of the absence of temporal correlation between ACEI therapy and the development of angioedema. Since angioedema may progress to upper airway obstruction, otolaryngologists must be aware of this association. Most importantly, late-onset angioedema should alert the clinician to discontinue the ACEI immediately to prevent further morbidity. This report presents an example of late-onset angioedema which was precipitated by taking a double dose of captopril incidentally. The case is discussed, and the literature, pathophysiology and treatment of angioedema are reviewed. PMID- 9210810 TI - The administration of balance testing is within the audiologists scope of practice. PMID- 9210811 TI - 1988 Audiology Practice Act. PMID- 9210812 TI - A bioelectric inverse imaging technique based on surface Laplacians. AB - A new approach is proposed to solve bioelectric inverse problems by employing the surface Laplacian of the bioelectrical potential. A theoretical investigation was conducted to test the feasibility of epicardial inverse imaging of cardiac electrical activity. A two-sphere homogeneous volume conductor model, where the inner sphere represents the epicardium and the outer sphere the body surface, was used. Radial and tangential current dipoles were used to approximate localized wavefronts propagating from the endocardium to the epicardium, and ectopic myocardial activities. The epicardial potential distribution was reconstructed from the body surface Laplacians with the aid of the Tikhonov zero-order regularization technique, which then was compared with the results obtained from the body surface potentials using the same regularization scheme. The two inverse solutions were compared qualitatively via visual inspection of the reconstructed epicardial potential maps, and quantitatively by examining relative errors and correlation coefficients between the "true" and the reconstructed epicardial potentials. Both qualitative and quantitative results indicate that the surface Laplacians play a positive role in improving the ill-posed nature of the bioelectric inverse problem, which would enhance our capability of reconstructing important epicardial events such as extrema in the epicardial potential distribution. The present theoretical study suggests that the Laplacian-based inverse imaging technique may have important applications to epicardial inverse imaging and other bioelectric inverse imaging. PMID- 9210813 TI - The Laplacian inverse problem of electrocardiography: an eccentric spheres study. AB - The eccentric spheres model of the heart-torso system is used to study the inverse problem of electrocardiography using measurements of the Laplacian of the body surface potential distribution. Electrical activity is simulated on the six main regions of the inner surface by considering a limited number of current dipoles placed within the inner sphere. The resulting outer surface potential and Laplacian distributions are then calculated in a forward sense. Varying amounts of random noise are added to these distributions before a zero-order Tikhonov regularization scheme is used to recover the inner surface potential distribution. Comparing the calculated and original inner surface distributions indicates that measurements of the outer surface Laplacian can more accurately reconstruct epicardial potentials than measurements of the outer surface potentials. These distributions are more accurate in that the extrema are placed very close to their original positions and are of nearly the same magnitude. Also, multiple extrema and high potential gradients are recovered. PMID- 9210814 TI - An approach for measuring ultrasonic backscattering from biological tissues with focused transducers. AB - When the standard substitution method is used with a focused transducer to measure the backscattering coefficient from biological tissues including blood, it yields erroneous results. Extending the backscattering measurements to frequencies beyond 15 MHz necessitates the use of focused transducers because of the worsened signal-to-noise ratio--caused by the increased attenuation and the smaller transducer aperture size--in order to make the measurements close to the transducer. An approach which allows the use of focused transducers in backscattering measurements has been developed. It has been used to measure the backscattering coefficient of red cell suspensions of hematocrit ranging from a few percent to 30% in the frequency range from 5 MHz to 30 MHz. The results at hematocrits below 20% agree well with those obtained with the standard substitution method, although they differ as the hematocrit is increased beyond 20%. The experimental results also show that the fourth-power dependence of backscatter on frequency is in general approximately valid for suspended erythrocytes of hematocrit between 6% and 30%. PMID- 9210815 TI - Comparison of impedance and inductance ventilation sensors on adults during breathing, motion, and simulated airway obstruction. AB - The goal of this study was to compare the relative performance of two noninvasive ventilation sensing technologies on adults during artifacts. We recorded changes in transthoracic impedance and cross-sectional area of the abdomen (abd) and rib cage (rc) using impedance pneumography (IP) and respiratory inductance plethysmography (RIP) on ten adult subjects during natural breathing, motion artifact, simulated airway obstruction, yawning, snoring, apnea, and coughing. We used a pneumotachometer to measure air flow and tidal volume as the standard. We calibrated all sensors during natural breathing, and performed measurements during all maneuvers without changing the calibration parameters. No sensor provided the most-accurate measure of tidal volume for all maneuvers. Overall, the combination of inductance sensors [RIP(sum)] calibrated during an isovolume maneuver had a bias (weighted mean difference) as low or lower than all individual sensors and all combinations of sensors. The IP(rc) sensor had a bias as low or lower than any individual sensor. The cross-correlation coefficient between sensors was high during natural breathing, but decreased during artifacts. The cross correlation between sensor pairs was lower during artifacts without breathing than it was during maneuvers with breathing for four different sensor combinations. We tested a simple breath-detection algorithm on all sensors and found that RIP(sum) resulted in the fewest number of false breath detections, with sensitivity of 90.8% and positive predictivity of 93.6%. PMID- 9210816 TI - Improvement of spatial resolution in surface-EMG: a theoretical and experimental comparison of different spatial filters. AB - The conventional bipolar surface electromyography (EMG) technique detects, due to its low spatial resolution, the superimposed electromyographic activity of a large number of motor units (MU's). In superficial muscles the isolated action potentials of the most superficial MU's can be recorded noninvasively by means of surface electrodes, if the method of spatial filtering, in connection with electrode arrays, is used. Up to now, only filters with an anisotropic transfer function have been used. As the surface potential distribution generated by the excitation of the MU's contains spatial frequencies in the anisotropic range of those filters, it can be assumed that isotropic spatial filters detect the single MU activity more effectively. In the present study, different isotropic and anisotropic filters have been compared by means of theoretical field simulations and experiments in volunteers. A tripole model for an excited MU was used as the basis for simulating the spatial extension of the filter response for each of the investigated filters. The spatial extension is an indicative of the spatial resolution. For the experimental validation, the total number of single motor units was not directly investigated, but the signal-to-noise ratio (SNR) has been determined. Therefore, the potential distribution generated on the skin surface during maximum voluntary contraction has been simultaneous spatially filtered with each of the investigated filters. The simulations show that an isotropic spatial filtering procedure reduces the spatial extension of the filter response and improves the spatial resolution of the EMG-recording arrangement in comparison to anisotropic spatial filters up to 30%. In other words, the spatial selectivity of the arrangement is increased. This improvement in the filter performance is more pronounced for MU's located close to the skin surface than for MU's more distantly located. Additionally, this theoretical improvement in selectivity depends on the direction of the excitation spread relative to the filter alignment. However, the investigations also show that isotropic filters offer an advantage, compared to anisotropic filters, only when the investigated MU is located extremely close to the filter input. The results of the simulations can be confirmed by the experimental investigations. An improvement of 11% in the SNR, relative to anisotropic spatial filters, can be established when using an isotropic spatial filter. This experimental improvement in selectivity is less than the theoretical improvement because the experimentally investigated MU's have less portion in the anisotropic range of the filters than the simulated one at best. PMID- 9210817 TI - Anatomical data fusion for quantitative reconstruction in myocardial tomoscintigraphy using a spline model of the thorax organs. AB - We present the fusion of anatomical data as a method for improving the reconstruction in single photon emission computed tomography (SPECT). Anatomical data is used to deduce a parameterized model of organs in a reconstructed slice using spline curves. This model allows us to define the imaging process, i.e., the direct problem, more adequately, and furthermore to restrict the reconstruction to the emitting zones. Instead of the usual square pixels, we use a new kind of discretization pixel, which fits to the contour in the region of interest. In the reconstruction phase, we estimate the activity in the emitting zones and also the optimum parameters of our model. Concentrating on the left ventricular (LV) wall activity, the simulation and phantom results show an accurate estimation of both the myocardial shape and the radioactive emission. PMID- 9210818 TI - Characterization of red blood cell aggregate formation using an analytical model of the ultrasonic backscattering coefficient. AB - Ultrasound backscattering is well adapted to study the red blood cell (RBC) aggregation phenomenon and growth of RBC aggregates since the backscattered ultrasonic intensity depends on the sixth power of the mean radius of the scattering centers when considered as spherical. Thus, small variations of aggregate size induce large variations of the backscattered intensity. From measurements of the ultrasonic backscattering coefficient (ultrasonic backscattering cross section per unit volume of suspension), an analytical model describing its variation versus time, for human aggregated red blood cells in sedimentation, is proposed. Results given by the model allow to define three phases in the phenomenon: 1) a starting phase characterized by a duration ts; 2) a stationary final phase beginning at time tf; 3) a growing intermediate phase characterized by its duration tf - ts. The analytical model has been applied to describe RBC aggregation in dextran 70,000 dalton of different concentrations, and at various hematocrits. Knowledge of the durations ts, tf and the maximum slope s of the curve during the intermediate phase, determined with the model, allows a means to study RBC aggregate growth. PMID- 9210819 TI - Light-guiding effect in a two-fluid model of laser angioplasty. AB - Mixing and optical characteristics of blood and optical fluid, utilized in laser angioplasty, are investigated with a two-fluid model. Transport equations are solved for the zone-averaged variables of each fluid with allowance for momentum transport at the interface. The predicted volume fractions of the fluids are used as weight functions to calculate the mixture refractive index. A set of light rays are traced through the fluids to the plaque, utilizing the mixture refractive index. The results indicate significant effect of flow characteristics on the focusing of the rays. PMID- 9210820 TI - Adaptive control of arterial blood pressure with a learning controller based on multilayer neural networks. AB - We discuss a two-model multilayer neural network controller for adaptive control of mean arterial blood pressure (MABP) using sodium nitroprusside. A model with an autoregressive moving average (ARMA), representing the dynamics of the system, and a modified back-propagation training algorithm are used to design the control system to meet specified objectives of design (settling time and undershoot/overshoot) and clinical constraints. The controller is associated with a weighting-determinant unit (WDU) to determine and update the output weighting factor of the parallel two-model neural network for adequate control action and a control-signal modification unit (CMU) to comply with clinical constraints and to suppress the effect of adverse noise and to improve the WDU performance. Extensive computer simulations indicate satisfactory performance and robustness of the proposed controller in the presence of much noise, over the full range of plant parameters, uncertainties, and large variations of parameters. PMID- 9210822 TI - Evaluation of antagonist coactivation strategies elicited from electrically stimulated muscles under load-moving conditions. AB - Muscle coactivation strategies that produce ankle dorsiflexion and plantar flexion were elicited by electrical stimulation of the tibialis anterior (TA) and soleus (SOL) muscles of the cat, and examined under several loading conditions. Four different load types were used: free-limb motion (no load), fly-wheel, and two pendulums, each with a different lever arm. Three types of coactivation strategies were considered. The first coactivation strategy consisted of antagonist activity that decreased as the agonist activity increased. The second strategy consisted of increasing antagonist activity with increasing agonist activity. And, in the third strategy, antagonist coactivation decreased at low force levels, then increased at high force levels. The three strategies were evaluated based on the joint angle's peak-to-peak movement and its ability to track a linear input command given by the correlation coefficient of the output signal versus linear input. Results showed that increasing antagonist activity resulted in decreasing peak-to-peak angle and a decreased signal tracking capability for each load condition. The latter, however, was not as obvious in the flywheel load (as compared with free-moving and pendulum conditions). A decreasing peak-to-peak torque for pendulum loads was also observed with increasing antagonist activity. In all loading conditions, maximal peak-to-peak angle and torque were present when a moderate degree of antagonist activity was engaged, and signal tracking capability improved with earlier engagement of the antagonist muscles. It is suggested that strategies using a combination of low level coactivation, as described in the physiological literature and previous functional electrical stimulation (FES) studies, could satisfactorily address the issues of controllability and efficiency while maintaining long-term joint integrity. PMID- 9210821 TI - The use of a PID controller to model vecuronium pharmacokinetics and pharmacodynamics during liver transplantation. Proportional-integral-derivative. AB - A four-phase proportional-integral-derivative (PID) controller was evaluated under the extremely unstable conditions of liver transplantation. Vecuronium was delivered to achieve 80%-90% neuromuscular blockade as measured by electromyogram (EMG). The first two controller phases delivered boluses and a constant infusion calculated to rapidly achieve setpoint, followed by a proportional-derivative (PD) phase at 35% from setpoint, and PID within 10% of the setpoint. During liver transplantation, the sources of system instability included large blood losses, temperature changes, and loss of hepatic drug metabolism during removal and replacement. During prolonged surgery, and when blood losses were not severe, the EMG remained within 10% of setpoint. Controller performance was more variable during system instability. Plasma sampling and two-compartment modelling of the infusion and response with a weighting factor for blood loss allowed estimation of the sources and degree of instability for improved design of future controllers. PMID- 9210823 TI - The influence of antagonist muscle control strategies on the isometric frequency response of the cat's ankle joint. AB - This study investigated the effect of various strategies to control the interaction between agonist and antagonist muscles on the frequency response of the isometric cat ankle joint actuated by the tibialis anterior (TA) and soleus (SOL) muscles. Some strategies were based on the physiologic need for increasing joint stability during forceful contractions; with these strategies, the proportional rate of physiologic antagonist activity was termed antagonist gain. Other strategies were based on the electrical stimulation literature, which advocates co-contraction at low force levels. The range of crossover of antagonist activity to the agonist's domain was termed overlap. Strategies consisting of 0%, 10%, and 20% antagonist gain were combined with 0%, 50%, and 100% overlap for a total of nine strategies. These were applied to the TA and SOL using sinusoidal input signals varying in frequency from 0.4 to 6 Hz. Gain and phase Bode plots were constructed through the use of the fast Fourier transforms (FFT's); and analysis of variance determined the significance of differences in gain and phase across frequencies. Best-fit models consisting of four poles and two zeroes were used to fit the experimental data and compared against an analytical model of muscles acting independently across the joint. Harmonic distortion was calculated to evaluate signal quality. It was found that changing the overlap and the antagonist gain produces significant changes in the dynamic response of the two-muscle joint system. The analytical approach to modeling such a system tends to consistently overestimate gain. It is suggested that signal quality is optimal when a moderate amount of antagonist gain (10%) is engaged, with overlap of 50% to smooth transitions between opposing movements. It is expected that this type of strategy will achieve optimum signal quality while preserving the long-term integrity of the joint. PMID- 9210824 TI - Using principal-components regression to stabilize EMG-muscle force parameter estimates of torso muscles. AB - Models for estimating muscle force from surface electromyographic (EMG) recordings require parameter estimates with low intertrial variability. The inclusion of multiple muscles in multivariate statistical models can lead to multicollinearity, especially when there are significant correlations between synergist muscles. One result of multicollinearity is that parameter estimates are very sensitive to changes in the independent variables. This study compared the parameter variability of multiple regression and principal-components regression techniques when applied to a six muscle EMG analysis of the lumbar region of the torso. Nine subjects participated. Twenty-three percent of the traditional multiple-regression parameters had incorrect signs, but none of the principal-components regression parameters did. The principal-components regression technique also produced parameter estimates having an order of magnitude smaller parameter variability. It was concluded that principal components regression is an effective method of mitigating the effect of multicollinearity in torso EMG models. PMID- 9210826 TI - What's up, doc? PMID- 9210825 TI - Adaptive reduction of heart sounds from lung sounds using fourth-order statistics. AB - When recording lung sounds, an incessant noise source occurs due to heart sounds. This noise source severely contaminates the breath sound signal and interferes in the analysis of lung sounds. In this paper, an adaptive heart-noise reduction method, based on fourth-order statistics (FOS) of the recorded signal, without requiring recorded "noise-only" reference signal, is presented. This algorithm uses adaptive filtering to preserve the entire spectrum. Furthermore, the proposed filter is independent of Gaussian uncorrelated noise and insensitive to the step-size parameter. It converges fast with small excess errors and, due to the narrow-band nature of heart noise (HN), it requires a very small number of taps. Results from experiments with healthy subjects indicate a local HN reduction equal to or greater than 90%. PMID- 9210827 TI - Biodegradable implants in hand and wrist fracture management. PMID- 9210828 TI - Vitamin D deficiency in two breast-fed infants. PMID- 9210830 TI - Heredity and systemic lupus erythematosus: dissecting a complex genetic disease. AB - The etiology of SLE appears to be exceedingly complex and possibly heterogeneous, with genetics and environment both making substantial contributions. A schematic representation of potential mechanisms is depicted in Figure 2. We may not fully understand the pathogenesis of this disease until we unravel the relative contributions of each component to the development of SLE. While genetic mechanisms involved in SLE remain obscure, we now have available elegant laboratory techniques for analysis of genetic loci as well as computer technology which permits simulation and analysis of the transmission of complex genetic traits among multiple families and demographic groups. What remains is the painstaking task of collecting families multiplex for SLE and analyzing multiple sets of clinical, serologic, and genetic data within and between these pedigrees. Such studies are currently underway and will hopefully increase understanding of this enigmatic and complex autoimmune disorder. PMID- 9210831 TI - Doctor Fabrique. PMID- 9210829 TI - Lipid screening and treatment by cardiologists have not improved. AB - Much effort by the national cholesterol education program (NCEP) and others have been made to induce physicians to screen for and treat lipid abnormalities in patients with coronary heart disease. We measured the effect of these efforts in a single group of cardiovascular specialists. We reviewed 20 percent of applicable patient records from 1987, 1989, and 1994 was performed to identify documented screening (cholesterol levels or lipid profiles) and treatment over 12 months after an index admission for coronary heart disease, along with a survey of physician acquaintance with NCEP guidelines, among the eight cardiovascular physicians. In the 160 patients with angina pectoris or myocardial infarction, total cholesterol levels were determined in 77-95 percent and lipid profiles determined in 2-11 percent. Treatment for cholesterol, greater than 150 mg/dl was initiated in 14-32 percent. These rates did not significantly improve over the study period. Yet, all physicians were acquainted with the NCEP and five of the eight perceived their screening and treatment to be more aggressive in 1994 than in 1987. Lipid screening and treatment by cardiovascular specialists have not improved despite copious supportive literature. Barriers other than lack of knowledge may impede implementation of this effective therapy. PMID- 9210832 TI - Managed care: it's been around a long time and it's not going away. PMID- 9210833 TI - Escherichia coli O157:H7 infections in Kansas, 1994-1995. PMID- 9210834 TI - Some thoughts on managed care. PMID- 9210835 TI - Unintentional injury death rates for Texas children drop, while intentional injury death rates rise. PMID- 9210837 TI - Review of Texas medical school finances. PMID- 9210836 TI - Identifying the cost of undergraduate medical education. PMID- 9210838 TI - Do not resuscitate. PMID- 9210839 TI - Payment hassles. PMID- 9210840 TI - Electronic filing. PMID- 9210841 TI - Medicine's mission. PMID- 9210842 TI - Medical history. PMID- 9210843 TI - Malpractice in the lower Rio Grande Valley. AB - Data from the National Practitioner Data Bank were examined to assess malpractice payments in the Lower Rio Grande Valley in comparison with those for Texas and the United States as a whole. The Valley was found to have a high rate of malpractice payments when considered on a per physician basis and on a per unit of utilization basis, but an average rate when considered on a population basis. A number of possible explanations (such as the "bad apple" theory, the "settlement for convenience" theory, the case distribution theory, the physician qualifications theory, and the foreign medical graduate theory) were examined to explain the Valley's observed rate of malpractice payments per physician. No explanations are particularly satisfactory. More research is needed, but indications suggest that the observed rate may be driven more by the activities of some attorneys than by the actions of Valley physicians. PMID- 9210844 TI - Medical malpractice arbitration: a primer for Texas physicians. AB - The medical malpractice crises and ensuing tort reform efforts, including methods of alternative dispute resolution (ADR), are generally reviewed. Arbitration in the context of medical malpractice is examined from the perspective of other states' experiences. Michigan has one of the nation's oldest medical malpractice arbitration programs, but it suffers from underutilization. California's experience derives from the use of arbitration in the managed care setting. While Texas has statutory provisions for medical malpractice arbitration, in light of public policy favoring ADR, the statute could be perceived as antipublic policy, resulting in underuse. The National Practitioner Data Bank also serves to discourage physician participation. Policy options are offered to address these concerns. PMID- 9210845 TI - Searching for preventable causes of infant mortality in Texas. AB - Texas' health objectives include reducing the infant mortality rate from 8.1 per 1000 live births in 1990 to no more than 7 per 1000 by the year 2000. To help target potentially preventable causes of infant mortality, we examined the underlying cause of each of the 2545 infant deaths that occurred in Texas during 1990. Potential existed for primary prevention of the underlying cause of at least some of 778 infant deaths and for secondary prevention by treating the underlying causes of 1127. The following appear to be the most promising targets and strategies for prevention: causes of neonatal mortality by improving the clinical care of high-risk newborns; sudden infant death syndrome by having infants sleep in the supine position; respiratory complications of preterm birth by preventive and therapeutic surfactant; death from bacterial infections by optimal antibiotic therapy; injuries and accidents by optimal parental supervision; preterm birth and adverse pregnancy outcomes by optimal prenatal, obstetric, and neonatal care; and neural tube defects by adequate paraconceptual folic acid. PMID- 9210847 TI - [Growth hormone after burns and multiple trauma. Immunologic effects of growth hormone]. PMID- 9210846 TI - Multicystic ovarian tumor weighing 156 lbs: the second largest tumor in Texas. AB - A woman's lifetime risk of developing an ovarian neoplasm is approximately 7%. A small subset of these neoplasms comprises the gigantic ovarian tumors (> 25 lbs) that often present challenging and difficult problems. We present a case of a 156 lb ovarian tumor with emphasis on the diagnostic and therapeutic approaches to successful management. PMID- 9210848 TI - [Evolution of the surgical treatment of chronic pancreatitis over the last 25 years]. AB - The author briefly reminds us of the physiopatholopy of chronic pancreatitis (CP) and of its two principal surgical therapies: the excision (mainly cephalic or caudal) and the derivations (essentially towards an excluded jejunal ring). In order to avoid such a mutilation, either pancreatic or jejunal for the treatment of obstructive "pancreatic lithiasis", the author proposes to classify the lesional repercussions in cavitary CP (in which the existence of a pseudocyst is dominant) and in parenchymatous CP (ensheating the ducts, that are more or less dilated). He infers from this the possibility of a treatment that is as physiological as possible, essentially by cystoduodenostomy (CD) with a tripod forceps, for CP with a dominant cavitary type (with pseudocysts showing a cephalic and/or a corporeocaudal localization) and by wirsungosphincteroclasia (W SC) for the CP with a parenchymatous prevalence. This therapeutic evolution, aiming at abandoning the classical operations of excision or derivation is based upon the author's experience acquired since 1970. During these 25 years, the author operated on 549 patients showing a CP with several severe evolutive complications. Beside 75 exopancreatic operations, the author performed 474 operations selectively concerning the pancreas: 245 excision operations and 228 derivation operations. Since the introduction, in October 1986, of the W-SC operation, among the 169 recent pancreatic operations for severe CP, only 10 exeresis operations were performed: 66 CD (41.5%) and W-SC 92 (57.9%), coupled in two thirds of the cases with a biliodigestive cholecystoplasty. The very encouraging results of this more physiological and non-mutilating treatment of severe CP justify, according to the author, forsaking the classical techniques of parenchym-exeresis or of derivation to an excluded ring an favour of a direct drainage into the duodenum both for a cavitary CP by CD as for a parenchymatous CP by W-SC. PMID- 9210849 TI - [Current tasks and structure of the Council on Public Health]. AB - The Council on Public Health (CPH) is a scientific council established in 1848 to assist the authorities in public and environmental health. The CPH has recently been restructurized mainly on the basis of changes in tasks related to obligations at the European level and to the integration of the National Council on Nutrition in the CPH. The CPH consists of 80 members; the CPH is subdivided in seven sections and one logistic unit. Several sections are further subdivided in subsections each of which deals with specific problems. The CPH is assisted by a scientific and an administrative secretariat. The activities of the CPH are illustrated with several examples of important subjects that were treated recently. PMID- 9210850 TI - [Treatment with the implantable cardioverter defibrillator (ICD)]. AB - Interventional techniques in clinical cardiology became important in rhythmology, because it is evident that antiarrhythmic drug therapy has several limitations, and side effects. The implantable cardioverter defibrillator is a very reliable tool in the therapy of ventricular fibrillation and ventricular tachycardia. Conversion of these arrhythmias is associated with the prevention of subsequent sudden cardiac death in patients who received implantation after cardiac arrest, not associated with myocardial infarction or metabolic disturbances. The exact place of the defibrillator in the treatment of ventricular tachycardia remains unsettled, as amiodarone, sotalol and catheter ablation are acceptable treatment modalities for some patients. Furthermore, associated antiarrhythmic drug therapy remains necessary in a large group of patients after implantation. The morbidity and mortality of the recently developed non-thoracotomy devices is low. The potential value of implantable defibrillators in the prevention of cardiac arrest in high-risk patients remains to be studied. PMID- 9210851 TI - Timing variability in children with early-treated congenital hypothyroidism. AB - This study reports on central and peripheral determinants of timing variability in self-paced tapping by children with early-treated congenital hypothyroidism (CH). A theoretical model of the timing of repetitive movements developed by Wing and Kristofferson was applied to estimate the central timekeeper (clock) and peripheral implementation (motor delay) variances from the variability in the response intervals. Before it is diagnosed and treated, CH is known to affect proper development of the cerebellum. If this would affect the time-keeper function of the cerebellum, it should be reflected by an increased central clock variability rather than by an increased peripheral motor-delay variability in terms of the Wing and Kristofferson model. Results of 46 children with early treated CH, differing in severity (21 severe, 25 mild), and 34 normal controls are reported. A refinement of the Wing and Kristofferson model is applied to estimate central clock and peripheral motor delay timing variability more precisely than has been done before. Results show that for children with early treated CH the estimate of the motor delay variance is four times higher than for the controls, while the estimate of the clock variance does not differ between the groups. It is concluded that motor problems in early-treated CH are associated with peripheral rather than with central timing deficiencies. PMID- 9210852 TI - General lexical slowing and the semantic priming effect: the roles of age and ability. AB - Older adults performed three lexical information-processing tasks approximately 1.3 times slower than young adults. Consistent with general lexical slowing, slopes of regressions based on individual subjects' RTs on two of the tasks (single lexical decision and category judgment) did not differ from slopes based on the third (double lexical decision) task. Moreover, slopes based on the single lexical decision and category judgment tasks accurately predicted the size of semantic priming effects on the third (double lexical decision) task. This was true for the older group as a whole, and also for subgroups of fast, medium and slow older adults, as well as for young adult subgroups. The size of the semantic priming effects for the fast old and slow young subgroups (who differed in age but not in processing speed) were approximately equal, consistent with the idea that the effect of age on priming is entirely attributable to slowing. Across all tasks, each old subgroup (fast, medium, or slow) showed the same degree of slowing relative to the corresponding young subgroup, so that the differences in RTs observed between subgroups in the young sample were magnified in the old sample. Taken together, the present findings suggest that ability-related differences in lexical processing speed may be functionally equivalent to age related differences and that both factors interact to determine performance on speeded lexical tasks. PMID- 9210853 TI - Adding new word associations to semantic memory: evidence for two interactive learning components. AB - The addition of newly learned word associations to semantic memory was investigated in three experiments. In these experiments word pairs were repeatedly presented as prime-target pairs in a lexical decision task. Performance on repeated pairs (both pre-experimentally associated and initially unrelated pairs) was compared to that on neutral pairs. In Experiments 1 and 2, effects of prior study (episodic priming) were observed but since this episodic priming effect was equal for both conditions it could not be concluded that the new associations has been added to semantic memory. In Experiment 3 some evidence was found that the newly learned word associations had been added to semantic memory. This occurred only after presenting the word pairs for several trials in paired-associate learning. The results are interpreted as supporting a model that distinguishes two memory components that mediate the effects of new learning, an episodic and a semantic one. PMID- 9210854 TI - Planning macroscopic aspects of manual control: end-state comfort and point-of change effects. AB - A recent emphasis in motor control research is the planning of macroscopic features and how variables such as efficiency and comfort influence the planning process. This paper extends the work by Rosenbaum and Jorgensen (1992) by further studying the end-state comfort effect. In the first experiment, participants picked up a dowel using an underhand or overhand grip and touched one end to a numbered target on the wall. The height of the #9 target was set at the height of participants' right shoulder. The second experiment involved awkwardness ratings. Participants touched the 14 targets with the dowel as well as with a small dumbbell and the comfort of the end position was rated on a seven-point scale. In the third experiment, participants moved a dumbbell to the targets in the same procedure as the first experiment. Overall, the probability analyses indicated that as the end-state comfort effect was magnified, the sequential effect vanished and a distinct point-of-change effect appeared. Optimization theory and the knowledge model readily explained the phenomena of the end-state comfort effect, the sequential effect, and the point-of-change effect. The present findings indicate that comfort has a powerful influence on the planning of motor performance. PMID- 9210855 TI - Defense Mechanism Test (DMT) and Kernberg's theory of personality organization related to adolescents in psychiatric care. AB - 75 adolescent psychiatric patients were diagnosed with the perceptual projective test the Defense Mechanism Test (DMT) and also according to Kernberg's theory of personality organization (PO). The test protocols were scored in respect of 130 DMT variables and analyzed by means of partial least squares (PLS) discriminant analysis. The objective was to try to separate the three types of PO, psychotic (PPO), borderline (BPO) and neurotic (NPO) by means of the DMT and also to compare the results with a similar study in adult psychiatric patients. The results showed that it is possible to separate significantly the three groups of PO. The BPO group seemed to be heterogeneous. The results were fairly similar to those obtained with adult psychiatric patients. The overall results supported the concurrent validity of Kernberg's theory of PO and for the DMT as well. The DMT seems to be a useful diagnostic method in respect of adolescent psychiatric patients. PMID- 9210856 TI - Social support after the loss on an infant child: a long-term perspective. AB - The article presents findings from a survey among 251 parents whose infant child had died. For most of the parents, the loss occurred several years ago. The survey assessed the amounts of instrumental, emotional and informational support received by these parents from various sources in connection with the death. The findings revealed that different sources provided different kinds of support. There was only one significant difference between bereaved males and females with regard to amount of support: females received more emotional support from their friends than males did. Furthermore, large amounts of support received by one spouse was associated with a similar level of support received by the other spouse. Social support in connection with the death was to some extent related to long-term psychological adaptation. Particularly support from neighbours and professionals was consistently associated with psychological adaptation. In general, however, the findings with regard to long-term effects of social support were ambiguous. PMID- 9210857 TI - Neuropsychological course after surgery for intracranial aneurysms. A prospective study and a critical review. AB - Previously, only three studies with representative samples of patients with ruptured intracranial aneurysms have provided detailed results of prospective, repeated, neuropsychological assessments after surgery. These studies apparently disagree with regard to occurrence of cognitive deficits and to degree of improvement between early and delayed follow-ups. The present paper attempts to analyze the conditions underlying these differences in results. As a first step in this analysis we present a comprehensive, prospective, neuropsychological investigation of a consecutive sample of 41 patients with rupture of a supratentorial aneurysm, assessed 4 and 12 months after surgery. It is concluded that a prorated course of improvement of a wide specter of psychological functions may be revealed, but that sensitive tests and large samples are needed to establish the range of deficits and improvements with time. Differences in patient selection with respect to severity of the acute clinical state and delayed deterioration apparently contribute importantly to the discrepance in previously reported outcome. PMID- 9210858 TI - Unemployment, coping and psychological distress. AB - This study addressed the role of coping style in anxiety and depression of unemployed people. Two-hundred thirty-three people checking in at unemployment services participated. They filled in Carver, Scheier and Weintraub's (1989) coping measure (COPE), the Hospital Anxiety and Depression scale (HAD), gave information as to age, duration of unemployment and their appraisal of their situation. Four secondary dimensions of COPE were used in data analyses. Multiple regression analyses were undertaken with anxiety/depression as dependent and the coping variables as independent variables, with background/appraisal variables entered first. Coping variables added to the prediction of anxiety and depression over and above background/appraisal variables. For women Focus on Emotion as well as Avoidance was related to higher anxiety/depression scores (p < 0.01), whereas Reappraisal was related to lower anxiety/depression (p < 0.05). For men only Avoidance was related to anxiety/depression (p < 0.01). More Avoidance co occurred with higher levels of anxiety as well as depression. The results are discussed with respect to possible intervention. PMID- 9210859 TI - Primary care for persons with disabilities. An overview of the problem. AB - This article outlines the ongoing health care needs of people with disabilities and how organized health care, particularly primary care, often fails to address these needs in a timely fashion. The article's central argument is that managed care and the ferment present in health care today present enormous opportunities for rehabilitation providers and others to develop creative solutions to address the shortcomings of the present health care system. PMID- 9210860 TI - Primary care for persons with disabilities. Framing the issues. PMID- 9210861 TI - Primary care for persons with disabilities. The public policy perspective. PMID- 9210862 TI - Primary care for persons with disabilities. The internal medicine perspective. PMID- 9210863 TI - Primary care for persons with disabilities. The family practice perspective. PMID- 9210864 TI - Primary care for persons with disabilities. The physiatrist's perspective. AB - It is difficult for many persons with physical disabilities to find and access primary health care. This article discusses this problem from the perspective of a physiatrist. It discusses the components of primary health care and then compares and contrasts the current status of the training, skills, and interest levels of generalist physicians and physiatrists in providing primary medical care for disabled persons. It includes a discussion of the artificiality of the specialist/generalist dichotomy and the concept of specialists providing true primary care to certain patient populations. General and personal strategies are suggested to influence and to change the health care system so disabled persons can find and have improved access to good primary health care. PMID- 9210865 TI - Primary care for persons with disabilities. The organized specialty medicine perspective. PMID- 9210866 TI - Primary care for persons with disabilities. The rehabilitation hospital perspective. PMID- 9210867 TI - Primary care for persons with disabilities. The Boston, Massachusetts model program. AB - Boston's Community Medical Group (BCMG) was among the first primary care group practices to provide community-based continuous primary care to people with major disabling conditions, the first to rely on nurse-practitioners as primary care givers, and one of the first to provide care on a prepaid capitated basis. With more than one thousand person-years' experience, BCMG has demonstrated that it is ethically and operationally feasible (1) to provide prepaid managed care to people with major disabling conditions; (2) to share financial risk for that care with providers; (3) to reinforce principles of independent living and consumer autonomy; (4) to assure high-quality clinical outcomes at reasonable costs. PMID- 9210868 TI - Primary care for persons with disabilities. A fragmented model of care for persons with spinal cord injuries. PMID- 9210869 TI - Primary care for persons with disabilities. The Rehabilitation Institute of Michigan Model Program. PMID- 9210870 TI - Nutritional support of the hospitalized patient. AB - Comprehensive care of patients in hospitals includes assessment of nutritional status and provision of appropriate support. This approach is facilitated by knowledge of the essential differences in metabolism between starved and stressed states. Nutritional assessment and care of patients in a hospital are based on answers to the following questions: Who gets it? When do they get it? How much do they get? What route is used to administer it? What kind do they get? What are common complications of enteral and parenteral support? What nutritional aspects are pertinent to common diseases? PMID- 9210871 TI - Differences between the pathogenesis of senile plaques and congophilic angiopathy in Alzheimer disease. PMID- 9210872 TI - In situ hybridization analysis of Girk2 expression in the developing central nervous system in normal and weaver mice. AB - A mutation in the gene Girk2 that encodes an inwardly rectifying potassium channel is the genetic defect causing the behavioral and pathologic abnormalities of the weaver mutant mouse. Of the pathologic abnormalities, the best studied is the neuronal degeneration that occurs in the cerebellar cortex and in the midbrain dopaminergic neurons. A detailed characterization of the topographic and temporal expression of Girk2 is fundamental to elucidate the mechanisms underlying neurodegeneration in these mutant mice. In this study we utilized in situ hybridization to determine the expression of Girk2 mRNA during prenatal and postnatal development in the murine central nervous system (CNS). Girk2 expression was seen in multiple regions of embryonic CNS including the cerebellum and midbrain. During postnatal development, the highest expression was seen in the cerebellum, midbrain and hippocampus. However, since the developing cerebellum undergoes significant neuronal loss due to the degeneration of granule cell precursors, Girk2 mRNA expression in this area decreases progressively. PMID- 9210873 TI - High-grade human brain tumors exhibit increased expression of myelin transcription factor 1 (MYT1), a zinc finger DNA-binding protein. AB - Detection and characterization of distinct central nervous system (CNS) tumor cell types is clinically important since distinct tumor types are associated with different prognoses and treatments. However, there is currently a lack of markers to identify certain glioma types and insufficient understanding as to which cells give rise to different glioma cell types. In the present study, biopsy specimens from human brain tumors were analyzed for expression of Myelin Transcription Factor 1 (MYT1) to explore the extent to which glioma cells reflect characteristic expression of MYT1 in developing glial progenitor cells. Immunostaining with an antibody against MYT1 revealed widespread immunoreactivity that was most prominent in high-grade oligodendrogliomas, astrocytomas, and mixed oligoastrocytomas as well as in a dysembryoplastic neuroepithelial tumor. MYT1 immunoreactivity in tumor regions generally correlated with the prevalence of cells exhibiting nuclear immunolabeling with an antibody against Ki-67, suggesting an association of MYT1 with cell proliferation that was also observed in normal adult human and rat brain in the germinal subependymal zone. The MYT1 immunoreactivity was frequently nuclear, appearing as dotted or punctate, but in some cases it was localized to the cytoplasm. In combination with histopathological studies and analysis of Ki-67 immunoreactivity, examination of MYT1 immunolabeling may provide additional information to aid in the detection and diagnosis of CNS tumors. PMID- 9210874 TI - Determination of p53 mutations, EGFR overexpression, and loss of p16 expression in pediatric glioblastomas. AB - Glioblastoma multiforme is a rare neoplasm in children and is often located infratentorially, particularly in the brainstem: Pediatric glioblastomas arise frequently (here 60%) outside the cerebral hemispheres. We investigated 20 pediatric glioblastomas for mutational inactivation of the p53 tumor suppressor gene, loss of p16 protein expression and overexpression of the epidermal growth factor receptor (EGFR). Mutations in the p53 gene were identified in 5/20 (25%) glioblastomas, 4 of which occurred in primary glioblastomas with a clinical history of less than 4 months and neither clinical nor histologic evidence of a less malignant precursor lesion. Loss of p16 expression was detected in 11/18 (61%) glioblastomas. Overexpression of the EGFR was infrequent (2/19, 11%) and included 1 tumor with a p53 mutation. Of 4 secondary glioblastomas that progressed from histologically diagnosed lower grade tumors, one contained a p53 mutation. Our results are at variance with similar studies in adult patients in which primary and secondary glioblastomas are characterized by EGFR overexpression and p53 mutations, respectively, suggesting that the evolution of pediatric glioblastomas follows different genetic pathways. PMID- 9210875 TI - BDNF and TrkB co-localize in CA1 neurons resistant to transient forebrain ischemia in the adult gerbil. AB - Delayed cell death of projection cells in the CA1 area of the hippocampus is produced in the adult gerbil following 5 minutes (min) of transient forebrain ischemia. Parvalbumin-immunoreactive local-circuit neurons are resistant to the ischemic insult. Brain-Derived Neurotrophic Factor (BDNF) immunoreactivity is localized in all neurons of the CA1 area in control gerbils. However, TrkB immunoreactivity is observed in a minority of BDNF-immunoreactive neurons in the CA1 area. The number of BDNF-immunoreactive cells in CA1 is dramatically reduced in ischemic gerbils as early as 24 h after ischemia, but the number of TrkB immunoreactive cells in the CA1 area is maintained following ischemia. Moreover, about 90% of BDNF-immunoreactive cells and about 85% of TrkB-immunoreactive cells in ischemic gerbils co-localize the calcium-binding protein parvalbumin. Finally, BDNF and TrkB are coexpressed in about 95% of CA1 neurons surviving the ischemic insult. These results indicate that a subpopulation of CA1 hippocampal neurons coexpressing TrkB, parvalbumin and BDNF is resistant to transient forebrain ischemia in the gerbil. These results also suggest that a subpopulation of CA1 hippocampal neurons in the gerbil hippocampus is endowed with a putative BDNF/TrkB autocrine regulatory loop that may be involved in both cell survival and synaptic remodeling of the damaged gerbil hippocampus following transient forebrain ischemia. PMID- 9210877 TI - A novel splice site associated polymorphism in the tuberous sclerosis 2 (TSC2) gene may predispose to the development of sporadic gangliogliomas. AB - The tuberous sclerosis 2 (TSC2) gene is thought to function as a growth suppressor in sporadic and TSC-associated hamartomas and tumors. Clusters of dysplastic glial cells are a common feature of cortical tubers and subependymal nodules in tuberous sclerosis patients. In an effort to identify TSC2 gene alterations in sporadic gliomas, we detected a novel polymorphism adjacent to the 3'splice site of intron 4. We evaluated the distribution of this variant allele in a series of 244 patients with glial tumors, including 55 gangliogliomas, 31 pilocytic astrocytomas (WHO grade I), 50 astrocytomas (WHO grades II and III), and 108 glioblastomas (WHO grade IV). The allelic distribution in the general population was estimated by examining 381 healthy blood donors. This rare allele appeared in the control population and in the patients with astrocytic gliomas with a virtually identical frequency (8.14%, and 8.20%, respectively). The frequency of the rare allele in gangliogliomas, however, was significantly higher (15.5%; p = 0.024). The fact that both gangliogliomas and cortical tubers in tuberous sclerosis contain neuronal and astrocytic elements and may resemble each other histologically suggests that the TSC2 gene may be involved in the development of these tumors. The rare allele of the TSC2 gene emerges as a candidate for a predisposing factor for the formation of sporadic gangliogliomas. PMID- 9210876 TI - A comparison of the predictive power for survival in gliomas provided by MIB-1, bromodeoxyuridine and proliferating cell nuclear antigen with histopathologic and clinical parameters. AB - The purpose of this prospective study of 65 patients was to compare side-by-side the predictive power for survival of (a) MIB-1, (b) bromodeoxyuridine (BUDR), and (c) proliferating cell nuclear antigen (PCNA). They were compared (a) with each other, (b) with several clinical predictors, and (c) with histopathologic grade under actual clinical biopsy conditions in a study of 1993 World Health Organization (WHO) grade II to IV adult supratentorial gliomas. There was a strong positive relationship between MIB-1 and BUDR by Spearman Rank correlation. In univariate analysis, MIB-1 (logrank p = 0.06) was more predictive of survival than BUDR or PCNA. Longer survivors were distinguished from others by the lowest MIB-1 labeling indices (LI < or = 2.5%) better than by the lowest histopathologic grade. However, histopathologic grades were highly predictive among the entire group (logrank p < 0.0001). Young age (p < 0.0001) and high Karnofsky performance status (p < 0.0001) were the clinical factors most predictive of longer survival. Female gender correlated with longer survival (logrank p = 0.02). In multivariate Cox proportional hazards models, age, Karnofsky performance status, and histopathologic grading remained statistically significant after full reduction of the model. We conclude that Ki-67 measured by MIB-1 monoclonal antibody was superior to other markers of proliferation. When all factors are considered simultaneously over all 3 grades of malignancy, greatest predictive power resides in histopathologic grade and clinical variables. MIB-1 is expected to be most important in cases where clinical or histopathologic factors are ambiguous or where they cannot be fully assessed. PMID- 9210878 TI - Heterozygous P0 knockout mice develop a peripheral neuropathy that resembles chronic inflammatory demyelinating polyneuropathy (CIDP). AB - Demyelinating peripheral neuropathies are clinically divided into inherited and acquired types. Inherited demyelinating neuropathies are caused by mutations in genes expressed by myelinating Schwann cells, whereas acquired ones, including chronic inflammatory demyelinating polyneuropathy (CIDP), are probably caused by autoimmune mechanisms. We find that heterozygous P0 knockout (P0+/-) mice develop a neuropathy that resembles CIDP. By one year of age, P0+/- mice develop severe, asymmetric slowing of motor nerves, with temporal dispersion or conduction block, which are features of acquired demyelinating neuropathies including CIDP. Moreover, morphological analysis of affected nerves reveals severe and selective demyelination of motor fibers, focal regions of demyelination, and inflammatory cells. These data suggest that immune-mediated mechanisms may contribute to the pathogenesis of the neuropathy in P0+/- mice. PMID- 9210879 TI - Characterization of diffuse axonal pathology and selective hippocampal damage following inertial brain trauma in the pig. AB - Dynamic deformation applied to white matter tracts is a common feature of human brain trauma, and may result in diffuse axonal injury (DAI). To produce DAI in an experimental model, we have utilized nonimpact inertial loading to induce brain trauma in miniature swine. This species was chosen due to its large gyrencephalic brain with substantial white matter domains. Twenty anesthetized (2% isoflurane) miniature swine were subjected to pure impulsive centroidal rotation 110 degrees in the coronal plane in 4 to 6 ms; peak accelerations ranged from 0.6 to 1.7 x 10(5) rad/s2. Seven days following injury, the brains were fixed (4% paraformaldehyde). Histopathologic examination was performed on 40 microns sections stained with cresyl violet (Nissl), antibodies targeting neurofilament (axonal damage), GFAP (astrocytes), and pig IgG (protein extravasation). Widespread multifocal axonal injury was observed in combination with gliosis throughout the brain, most commonly in the root of gyri and at the interface of the gray and white matter. Very little vascular disruption was noted in regions of axonal injury. Neuronal damage was primarily found in the CA1 and CA3 subfields of the hippocampus. These results suggest that this model is clinically relevant and useful for evaluating mechanisms of inertial brain trauma. PMID- 9210880 TI - Reduced expression of the P2 form of the gap junction protein connexin43 in malignant meningiomas. AB - Neoplastic transformation is often associated with aberrant gap junctional intercellular communication. We assessed mutations and expression of the connexin43 (Cx43) gene in 49 intracranial meningiomas. SSCP analyses followed by direct DNA sequencing showed GCG-->GTG (Ala-->Val) transition mutation in codon 253 of the cytoplasmic carboxyl terminal of the Cx43 gene in 1 of 31 (3%) benign meningiomas and 1 of 14 (7%) anaplastic meningiomas. The same base change was present in normal tissue from these patients and also in 4 of 80 (5%) DNA samples extracted from lymphocytes of healthy Europeans, suggesting that this constitutes a newly identified Cx43 polymorphism. Western blot analyses showed expression of phosphorylated P1 (45 kD) and P2 (47 kD) Cx43 as well as the unphosphorylated form (42 kD) in 11 of 14 (79%) benign meningiomas. In contrast, the P2 form was not detectable in the majority (7 of 9; 78%) of atypical and anaplastic meningiomas. Since the presence of the P2 form is often associated with optimal function of the Cx43, these results suggest that loss or impaired gap junctional cell to cell communication may be associated with meningiomas displaying more rapid growth and a less favorable prognosis. PMID- 9210881 TI - Diabetic neuropathy. PMID- 9210882 TI - Clinical evaluation of anti-tumor effects of Lentinan combined with chemotherapy in the treatment of various malignancies. PMID- 9210883 TI - Management of hepatocellular carcinoma: long-term outcome in 2639 cases. AB - This paper reports the progress of management in 2639 patients with pathologically proven primary liver cancer (PLC) over the past three decades, and the factors improving long-term outcome. The 5-, and 10-year survival after resection of PLC was 45.9% and 34.8%, respectively, for the whole series (n = 1826), and 61.3% and 45.7%, respectively, for patients with small PLC (< = 5 cm, n = 645). The 5-year survival after cryosurgery was 37.9% for the whole series (n = 191), and 53.1% for patients with small PLC (n = 56). The 5-year survival of 73 patients receiving sequential resection after cytoreduction therapy was 67.8%. The 5-year survival after re-resection for recurrence tumor (n = 148) was 34.5%; 239 patients survived more than 5 years; 124 of these patients (51.9%) were small PLC, and 63 patients survived more than 10 years. Encouraging changes in the prognostic pattern were observed when the PLC data of 1958-1970 (n = 178), 1971 1982 (n = 582) and 1983-1994 (n = 1879) were compared; the 5-year survival being 4.8%, 11.2% and 45.4%, respectively, and the 10-year survival being 4.2%, 7.5% and 34.6%, respectively. Some aspects to prolong survival further were discussed. PMID- 9210884 TI - Therapeutic strategies for early gastric cancer. PMID- 9210885 TI - Free ileocolon transfer after hypopharyngo-laryngo-cervical esophagectomy for speech rehabilitation. AB - Patients with carcinoma of the hypopharynx or the cervical esophagus usually undergo total laryngectomy with hypopharyngo-esophagectomy and, consequently, lose the power of speech. Therefore, a reconstruction method which would enable speech rehabilitation is desirable following this type of procedure. Free ileocolon transfer consists of colo-esophagostomy, pharyngo-colostomy, ileo tracheotomy, vascular anastomosis under a microscope, and permanent tracheostomy. In this method, the colon functions as the alimentary tract, while the ileum and ileocecal valve can produce sounds. We evaluated the results in live patients with free ileocolon transfer following hypopharyngo-laryngo-cervical esophagectomy. Although there was no case of direct operative death, one patient died from cancer progression. Four patients are still alive at from 18 to 36 months after operation. Postoperatively, stenosis of the colo-esophaogostomy appeared in two patients, but there was no case of anastomotic breakdown or any other major complication. Speech rehabilitation was good, and there was no misswallowing into the airway. We think that free ileocolon graft is one of the more preferable procedures following hypopharyngo-laryngo-cervical esophagectomy. PMID- 9210886 TI - Treatment results of radiotherapy for cervical cancer--experience of Tokyo Women's Medical College over a 25-year period. PMID- 9210887 TI - Arterial infusion chemotherapy for peritoneal and liver metastasis in gastric cancer. PMID- 9210888 TI - The role of chemotherapy in the treatment of head and neck cancer. PMID- 9210889 TI - Individualized chemotherapeutic regimen for each histological subtype of ovarian carcinoma. AB - Ovarian carcinoma can be classified into the following distinct two groups in terms of chemosensitivity. The chemosensitive group includes serous, endometrioid, transitional cell carcinoma (TCC), and undifferentiated carcinoma, with a response rate to CDDP-based chemotherapy (CTX) being approximately 80 to 90%. Even within this chemosensitive group, the mode of the response to chemotherapy may vary by the histologic subtype or the grade of differentiation. The chemo-resistant group consists of mucinous and clear-cell adenocarcinoma (CCA), with a response rate to CDDP-based regimen being less than 5% or none. Especially, for pure CCA of the ovary, no one case has ever been reported to have shown an appreciable response to a CDDP-based regimen. As a result, advanced patients with mucinous carcinoma or CCA of the ovary not amenable to a complete surgery have a poor prognosis. The author developed a promising new regimen consisting of CPT-11 with mitomycin-C based both on in vitro and in vivo chemosensitivity tests. Thus, an individualized regimen may be required for both CDDP-sensitive and resistant diseases. PMID- 9210891 TI - Development of new anticancer drugs in Japan--tubulin-interacting agents. PMID- 9210892 TI - Cancer therapy and apoptosis. PMID- 9210890 TI - New anticancer drugs in Europe. PMID- 9210893 TI - New drugs for the treatment of lung cancer. The Tokyo Cooperative Oncology Group. PMID- 9210894 TI - Chemotherapy for gastric cancer in Japan. PMID- 9210895 TI - Three decades' experience in surgery of hepatocellular carcinoma. AB - In the author's institution, 2254 patients with hepatocellular carcinoma (HCC) have been treated during 1958-1994. The overall 5-year survival increased from 5.4% (1958-1970), to 11.9% (1971-1982), to 46.2% (1983-1984), which correlated well with the increasing proportion of small HCC in the series (2.6%, 12.1%, and 33.4%, respectively); with the increasing percentage of limited resection (3.1%, 32.2%, and 58.3%); with the increasing number of re-resections for recurrence (0, 27, and 114 patients); and with the increasing number of second stage resections (0, 5, and 67 patients). In our institution, surgical approaches that resulted in significantly prolonging survival included: small HCC resection, re-resection, and cytoreduction followed by sequential resection for initially unresectable HCC. Experience in these 3 aspects suggests: (a) Small HCCs are mainly found by screening using AFP and ultrasonography (US) in a high risk population, and limited resection is the best treatment in patients with compensated liver cirrhosis, the 5-year survival after resection being 62.9% (n = 549). (b) Postoperative monitoring using AFP/US every 2-3 months for 5-10 years after curative resection is needed to detect subclinical recurrence. Limited re resection is indicated for liver recurrence less than 3 nodules, and lung lobectomy is of proven merit to prolong survival for solitary lung metastasis. Re resection of subclinical recurrence has resulted in a 10-20% further increase in 5-year survival after curative resection. (c) Palliative surgery other than resection such as hepatic artery ligation (HAL) and cannulation with arterial infusion (HAI), cryosurgery, etc. are superior to palliative resection with residual cancer. (d) Cytoreduction and sequential resection have provided hope for localized unresectable HCC, particularly in the right cirrhotic liver. Multimodality combination treatments such as HAL+HAI+radioimmunotherapy/regional radiotherapy are acceptable cytoreductive therapies. Repeated transcatheter hepatic arterial chemoembolization (TACE) is an alternative nonsurgical approach. Sequential resection is important to eradicate residual cancer after cytoreduction. The 5-year survival of 72 patients with cytoreduction and sequential resection for initially unresectable HCC was 62.1% and resulted in improving 5-year survival in the entire series of unresectable HCC over the 3 periods from 0% to 7.4% to 25.7%, respectively. However, multicentric origin and tumor invasiveness are two major targets to be studied in the control of recurrence and metastasis. PMID- 9210896 TI - Surgical treatment for hepatocellular carcinoma. PMID- 9210897 TI - Chemoembolization for small hepatocellular carcinoma. PMID- 9210898 TI - Dose intensification and breast cancer. PMID- 9210899 TI - ACOS presentations for publication in the Japanese Journal of Cancer and Chemotherapy. PMID- 9210900 TI - Cytokine therapy for hematological malignancies. AB - Recently various cytokines have been introduced into the clinic and have played important therapeutic roles in the treatment of hematological malignancies. Among these cytokines, we have focused on interferon (IFN) and granulocyte (G) or granulocyte-macrophage (GM) colony stimulating factor (CSF), which are currently the most useful cytokines in this review. IFN-a is one of most useful and wide ranging antitumor agents in hematological malignancies. The most striking effects have been studies in chronic phase CML. Cytogenetic responses are seen in 30-40% of the treated patients, and a complete cytogenetic response can be seen in about 10%. Long-term survival can be expected in these patients. Considering the risk of graft-versus-host disease-associated mortality in allogeneic bone marrow transplantation, the most appropriate category of treatment is difficult to determine in IFN-responsive patients. Elucidation of the antitumor mechanism of IFN, as a prototype for other biological response modifiers, may revolutionize cancer treatment. G- and GM-CSF (CSFs) have reduced the duration of neutropenia, incidence of infectious episodes and days of hospitalization following cancer chemotherapy or stem cell transplantation. CSFs have also been used to mobilize peripheral blood stem cells and to increase the dose intensity of chemotherapeutic agents. Leukemic cells from many patients with acute myelogenous leukemia (AML) have surface receptors for CSFs and may proliferate in response to CSFs. However, several randomized studies showed that CSFs can be used safely and effectively in augmenting neutrophil recovery in patients with AML when given after induction chemotherapy. Various trials have been conducted to sensitize leukemic cells by CSFs, making them more susceptible to chemotherapy; but no convincing evidence has been obtained. PMID- 9210901 TI - Modulation of 5-FU and its related compounds. PMID- 9210902 TI - Taking chemotherapy from random to rational with the histoculture drug response assay. PMID- 9210903 TI - Selection of adjuvant chemotherapy for gastric cancer using objective criteria. AB - Precise prediction of recurrence risks is of importance in the selection of adjuvant chemotherapy. A proportional hazards regression analysis was performed to seek objective criteria for 234 gastric cancer patients. The analysis showed that a high level of carcinoembryonic antigen (CEA) in peritoneal washing (100 ng/g protein) was an independent risk factor for peritoneal recurrence, and a high MMP9 (92 KD type IV collagenase) level in serum was a powerful indicator of hematogenous recurrence. In addition, the PHREG analysis in 1453 gastric cancer patients showed that MF therapy (mitomycin C i.v. + fluoropyrimidine derivatives) and PF therapy (Cisplatin i.p. + fluoropyrimidine derivatives) might be effective in the prevention of hematogenous and peritoneal recurrence after a curative operation, respectively. Based on these findings, a recurrence type-oriented adjuvant chemotherapy, i.e., PF therapy for high CEA group and MF therapy for high MMP9 group, was designed. The retrospective analysis showed significant survival benefit in stage III gastric cancer patients who underwent a curative operation. The 5-year survival rate was 67%, while that of the historical control was 46%. Prediction of recurrence type using CEA levels in peritoneal washings and MMP9 levels in sera may be of value in the selection of a proper population of patients for PE or MF therapy after gastrectomy. The selection system of adjuvant chemotherapy using such objective criteria may improve the prognosis of gastric cancer patients and keep their good quality of life through the selection of proper candidates and by eliminating patients in no need of therapy. PMID- 9210904 TI - Pre- and/or post-operative immunochemotherapy for advanced digestive cancer. PMID- 9210905 TI - New strategy of bio-chemoprevention on recurrence of superficial bladder cancer based on a hypothesis of the mechanism of recurrence. AB - There are theoretical limits to the efficacy of intravesical chemotherapy for prevention of tumor recurrence after transurethral resection of a superficial bladder cancer. Our multi-institutional studies revealed that the direct efficacy of BCG, intravesical instillation for treatment of an existing tumor is very promising. This efficacy persisted over a long period of time, and the subsequent recurrence rate was markedly reduced. Bladder cancer, sometimes earlier known as an occupational disease, might be related to unknown chemical carcinogens. Since enterobacterias are thought to produce carcinogens and mutagens, including nitroso-compounds in the intestinal tract, BLP (lactobacillus casei preparation), treatment may suppress the production of such compounds by altering the intestinal flora. Preclinical studies have demonstrated that BLP suppresses the development of bladder cancer induced by N-butyl-N-(4-hydroxy-butly)-nitrosamine in mice and rats. A double-blind clinical trial recently revealed that BLP was effective for preventing the recurrence of superficial bladder cancer. Bropirimine, a interferon inducer, is now an internationally developing agent for superficial bladder cancer, which is discussed on the basis of Japanese phase II trial data. PMID- 9210906 TI - mRNA expression, measured by quantitative reverse transcriptase polymerase chain reaction, of five putative drug resistance parameters, in normal and leukaemic peripheral blood and bone marrow. AB - Using a quantitative reverse transcriptase PCR assay, the mRNA expression of five putative drug resistance-related genes were assessed in normal peripheral (n = 14) and bone marrow (n = 4) mononuclear cells from healthy donors and patients with acute myeloid leukaemia (n = 11). The mRNA levels of MDR1, the multidrug resistance-associated protein and glutathione-S-transferase pi were equally expressed in both compartments. Bcl-2 mRNA was slightly higher in the leukaemic marrow samples. However, topoisomerase II alpha mRNA levels were found to be much higher in normal and leukaemic marrow cells compared to peripheral blood (p < 0.01), which may, in part, reflect the different proliferation pattern of the mononuclear cells in the two compartments. Such findings could be important for researchers using bulk assays in a mix of samples from peripheral blood or bone marrow to investigate prognostic factors in patients with leukaemia. PMID- 9210907 TI - Apoptosis and secondary necrosis of lymphocytes in culture. AB - It has been reported that cultured peripheral B lymphocytes of chronic lymphocytic leukemia (B-CLL) patients show a high degree of apoptosis (programmed cell death). Till now, no data exist about the occurrence of in vitro apoptosis of normal B and T cells. We measured the amount of apoptosis and secondary necrosis (type 2 necrosis) in B-CLL lymphocytes and in normal peripheral B and T lymphocytes in culture. Observations were made on B-CLL lymphocytes and on normal B and T cells purified by immunomagnetic cell sorting. Apoptosis and secondary necrosis were measured using a recently described sensitive flow-cytometric assay, probing simultaneously for cell surface exposure of phosphatidylserine with the use of FITC-labeled annexin-V and for cell membrane integrity as demonstrated by the exclusion of propidium iodide. The degree of in vitro apoptosis and secondary necrosis of normal B cells appears to be higher than that of normal T cells, and even higher than that of B-CLL cells. The results indicate that cultured mature circulating normal B lymphocytes exhibit a higher cell death rate than normal T cells and B-CLL lymphocytes. PMID- 9210908 TI - Complex effects of interleukin 6 on clonogenic blast cell growth in acute myeloblastic leukemia. AB - The present in vitro study shows how interleukin (IL)-6 modulates clonogenic blast cell growth in complex ways in acute myeloblastic leukemia when used either as a single factor or in different hematopoietic growth factor combinations. In the presence of IL-6, the colony numbers in culture assay decreased to 50 +/- 29% from the basal values (p < 0.001) in 10 cases and increased to 384 +/- 278% of the basal values (p < 0.01) in 5. The inhibitory effect of IL-6 on blast cell colony formation was retained when IL-6 was combined with granulocyte colony stimulating factor, but was lost if IL-6 was used in combination with mast cell growth factor, IL-3, granulocyte-macrophage colony stimulating factor, or IL-4. The stimulatory effect of IL-6 was diminished in the presence of granulocyte colony stimulating factor, but preserved in the presence of other growth factor combinations. IL-6 had a neutral effect on colony growth in 7 cases with acute myeloblastic leukemia. In these cases, however, IL-6 stimulated significantly clonogenic cell growth if combined with mast cell growth factor or granulocyte macrophage colony stimulating factor. PMID- 9210909 TI - Resistance to activated protein C and Arg 506 Gln factor V mutation are uncommon in eastern Asian populations. AB - We investigated the prevalence of the factor V (FV) Arg 506 Gln mutation in healthy subjects from three eastern Asian countries (Japan, n = 270; China, n = 113; and Korea, n = 93) and in 26 Japanese patients showing venous thromboembolic events. The patients were also examined for activated protein C (APC) resistance by using the Coatest APC resistance kit. The FV mutation was investigated by polymerase chain reaction and restricted enzyme digestion with MnlI RFLP assay of the FV gene. None of the patients showed APC resistance, while all subjects examined were homozygous for Arg at position 506 of the FV gene. Our results imply that FV mutation and APC resistance contribute little to venous thrombotic diseases in eastern Asia. PMID- 9210910 TI - Characteristics of a cell line established from a case of acute megakaryoblastic leukemia. AB - Acute megakaryoblastic leukemia is uncommon and comprises about 5% of acute nonlymphoid leukemias in the French-American-British classification. Cell lines from such leukemias are relatively rare with only about 8 reported in the literature. We established a cell line from a case of acute megakaryoblastic leukemia arising in a 2-year-old child. Surface marker studies of the cells confirmed their megakaryoblastic nature, with 54% of the cells being CD61 positive and peroxidase and esterase negative. The cells had a doubling time of 72 h. Emperipolesis (a phenomenon in which a cell, usually a lymphocyte or neutrophil, enters another cell, moves about and leaves without undergoing phagocytosis) of one blast into another, larger one, was occasionally seen, and a review of the original bone marrow specimen also showed emperipolesis of neutrophils into the megakaryoblasts. The cells responded to interleukin 3 and were inhibited with all-trans-retinoic acid. The karyotype of the cells was the 46,XX,-16 with a marker chromosome. The marker chromosome is possibly chromosome 16 with a small segment of a chromosome translocated to the terminal portion of chromosome 16. PMID- 9210911 TI - Presence of Epstein-Barr virus genome in the bone marrow of patients with hematopoietic malignancies. AB - The Epstein-Barr virus (EBV) genome was detected by polymerase chain reaction (PCR) in mononuclear cells from bone marrows with diverse types of hematopoietic malignancies. Viral repeated sequences (BamHI-W region) were detected in 42 of 82 (51%) hematopoietic malignancies, including polycythemia vera, but not in nonneoplastic cases. EBV-positive cases were found to consist of various histological types. We did not detect any EBV PCR product in the peripheral blood. The EBV BamHI-Y, -H region, encoding EBV nuclear antigen 2 DNA, which is a single-copy gene in the viral genome, was detected in only 13 of 42 BamHI-W positive cases, suggesting that the copy number of the EBV genome differed in each case. In all cases, the PCR band was verified by Southern blot hybridization using specific EBV probes. Whether the infected virus is an etiologic agent of the malignancy or merely a latent infection cannot be determined by the PCR assay performed under these conditions. These results, however, suggest that a novel form of EBV latent infection is present in the bone marrow of patients with hematopoietic malignancies. PMID- 9210912 TI - A patient with basophilic-eosinophilic myeloproliferative disorder showing monosomy 7 and hyperhistaminemia. AB - We encountered a male patient with marked basophilia and eosinophilia complicated by anemia, thrombocytopenia, myelofibrosis, and hyperhistaminemia. Since morphological abnormalities were unclear and since chromosome analysis showed 45,XY,-7, a diagnosis of basophilic-eosinophilic myeloproliferative disorder was made. After administration of prednisolone and cytarabine ocfosfate, basophil and eosinophil levels decreased, but blasts transiently appeared in the peripheral blood. Chromosome analysis performed at the time of appearance of blasts showed a clone with 45,XY,-7,del(16)(q22). Subsequently, pancytopenia developed, after which white blood cell count and its classification were normal, as were chromosome findings. In this patient, monosomy 7 seemed to have induced myeloproliferative disorder with basophilia and eosinophilia, and del(16)(q22) may have enhanced the eosinophilia. PMID- 9210913 TI - Acute cardiac tamponade associated with pericardial extramedullary hematopoiesis in agnogenic myeloid metaplasia. AB - A 67-year-old man was diagnosed with agnogenic myeloid metaplasia. Several months later, he presented with dyspnea, malaise and chest pain. A pericardial effusion was evident and a pericardial biopsy was consistent with extramedullary hematopoiesis. PMID- 9210914 TI - Flare-up of squamous cell carcinoma of the skin following fludarabine therapy for chronic lymphocytic leukemia. AB - We present a 72-year-old patient with chronic lymphocytic leukemia (CLL). About a year following therapy with chlorambucil and prednisone, he suffered from anemia, thrombocytopenia and organomegaly. The patient received fludarabine with a favorable response. Concomitantly with the clinical improvement of the CLL there was a remarkable flare-up of scalp squamous cell carcinoma (SCC) lesions, initially noted 4 years previously. The lesions were multiple and grew rapidly. Fludarabine depresses the T lymphocyte population, cells that play a pivotal role in the regression of the SCC. We suggest, that the flare-up and exacerbation of the SCC lesions of the patient were triggered by the fludarabine therapy. PMID- 9210915 TI - Pulmonary hypertension in two patients with type I Gaucher disease while on alglucerase therapy. PMID- 9210916 TI - SfaNI polymorphism distinguishes the alleles of the glycophorin A locus that determine the MN blood group. PMID- 9210917 TI - Identification of hematopoietic stem cells by the SE-9000 automated hematology analyzer in peripheral blood stem cell harvest samples. PMID- 9210918 TI - A simple method for evaluation of latex phagocytosis by rat peritoneal macrophages. PMID- 9210919 TI - The association of myeloproliferative and lymphoproliferative diseases. PMID- 9210920 TI - Pestiviruses--taxonomic perspectives. AB - The history of pestivirus taxonomy is surprisingly consistent: almost 30 years ago it was recognized that pestiviruses are structurally akin to flaviviruses, and recent nucleotide sequence data have confirmed this resemblance at the level of genome organization. For other enveloped positive stranded RNA viruses with ikosahedral nucleocapsids e.g. equine arteritis and lactate dehydrogenase virus of mice a taxonomic dilemma is encountered; while virions resemble ("non arthropod-borne") togaviruses, the replication via a nested set of subgenomic RNAs is corona and torovirus-like. Pestiviruses, flaviviruses and the hepatitis C virus group have been assigned generic status in the Flaviviridae family. PMID- 9210921 TI - Molecular characterization of hog cholera virus. AB - An efficient tissue culture system was established which allowed to obtain substantial quantities of hog cholera virus (HCV) from the cell free tissue culture supernatant. After preparation of viral RNA and cDNA synthesis, the complete HCV genome was cloned and sequenced. Comparison with published BVDV sequences revealed a surprisingly high homology between HCV and BVDV at both the nucleotide and the amino acid level. In addition host cellular sequences were identified in BVDV genomes. The genomic localization of HCV glycoproteins was determined by the use of sequence specific antisera directed against bacterial fusion proteins. The order on the HCV genome was determined as follows: N-gp44/48 gp33-gp55-C. HCV gp33 and HCV gp55 were shown to be intracellularly linked by disulfide bridges. A cDNA fragment covering the genomic region that encodes the structural proteins of HCV was inserted into a vaccinia recombination vector. Expression studies with vaccinia/HCV recombinants led to identification of HCV specific glycoproteins which migrated on sodium dodecyl sulfate-gels identically to glycoproteins precipitated from HCV-infected cells. The vaccinia virus/HCV recombinant that expressed all four structural proteins induced virus neutralizing antibodies in mice and swine. After immunization of pigs with this recombinant virus, full protection against a lethal challenge with HCV was achieved. A construct that lacked most of the HCV gp55 gene failed to induce neutralizing antibodies but induced protective immunity. PMID- 9210922 TI - Bovine viral diarrhea virus genomic organization. AB - In previous work, we developed a preliminary description of the genetic organization of the prototypic pestivirus bovine viral diarrhea virus (BVDV). In order to refine this genetic map and to further elucidate the gene products and expression strategy of this virus, we have generated a broad panel of sequence specific antibody reagents. Use of these reagents not only allowed the identification of several previously undescribed viral polypeptides, but when used in in vivo pulse-chase experiments, they identified precursor polyproteins and processing intermediates. Data generated from these studies provide a more accurate and complete view of viral gene organization, as well as insight into several aspects of protein processing and the gene expression strategy employed by this pestivirus. These experiments also revealed varying stability and turnover rates for the mature BVDV proteins. These latter results have implications for the functional roles of certain gene products. PMID- 9210923 TI - Bovine viral diarrhea virus proteins and their antigenic analyses. AB - Bovine Viral Diarrhea Virus (BVDV) polypeptides present in infected cells are the result of the processing of the polyprotein translated from the large single open reading frame of the BVDV genomic RNA. The presence of these proteins in infected cells was studied by radiolabeling under hypertonic conditions and with the aid of radioimmunoprecipitation. The genomic mapping of these polypeptides suggests a complex pattern of processing which involves cellular and viral proteases. The consistent absence of 80k in noncytopathic isolates of BVDV suggests that the processing of the viral polyprotein is different in cytopathic and noncytopathic biotypes of BVDV. The antigenic structure of BVDV was studied with a panel of monoclonal antibodies (MABs) prepared against the Singer isolate of BVDV. Neutralizing MABs were found to bind the 56-58k polypeptide, providing evidence that this glycoprotein is present on the surface of the virion and carries neutralization epitopes. Antigenic analyses with the panel of MABs reveals extensive antigenic heterogeneity among BVDV field isolates. MABs were used to determine the frequency of neutralization escape mutants in stocks of BVDV. Plaque-purified BVDV stocks contain neutralization escape mutants with a frequency of 10(-2.47). PMID- 9210925 TI - BVD monoclonal antibodies: relationship between viral protein specificity and viral strain specificity. AB - Seventeen monoclonal antibodies raised against bovine viral diarrhoea virus were divided into three groups on the basis of radioimmunoprecipitation results. Seven monoclonal antibodies precipitated a polypeptide of 80kD and defined four domains, all of which showed considerable conservation amongst the 180 pestivirus strains and isolates examined. Nine monoclonal antibodies, including six with virus neutralizing activity, precipitated a 53kD polypeptide and all appeared to be directed towards a single domain of clustered epitopes. Several of these epitopes were present in many ruminant virus strains and isolates, but not in hog cholera viruses. A single monoclonal antibody precipitated a 48kD polypeptide, defining an epitope that was also present on many ruminant viruses, but not hog cholera viruses. Most pestiviruses from cattle and some from sheep shared a number of epitopes located on three different proteins. PMID- 9210926 TI - Correlation of bovine viral diarrhoea virus induced cytopathic effects with expression of a biotype-specific marker. AB - The purpose of this study was the identification antigenic differences between cytopathic (cp) and noncytopathic (ncp) bovine viral diarrhoea viruses (BVDV). Cells infected with 19 strains of each viral biotype were analyzed for reactivity with the monoclonal antibody (mab) BVD/C38. Reactivity was examined using an enzyme immunoassay on fixed infected monolayers of fetal calf kidney cells. In the majority of cases, the mab discriminated between cells infected with each of the two viral biotypes. Three reactivity patterns could be distinguished. Most cpBVDV strains yielded monolayers where 80-100% of infected cells reacted with the mab. Most of the ncpBVDV infected cells showed either no reaction, or only single cells of foci were stained. However, about one third of either cp- or ncpBVDV strains tested yielded infected monolayers where 30-50% of the cells reacted with the antibody. Cell damage other than the typical cytopathic effect might be responsible for the BVD/C38 reactivity of cells infected with BVDV. In addition, it was analyzed whether the antigenic marker associated with cpBVDV was expressed in cells infected with viral isolates from 21 animals with clinical mucosal disease. In 14 cases cpBVDV was isolated and the antigenic marker was found throughout. In seven cases ncpBVDV was cultivated and the antigenic marker was detected in four isolates. PMID- 9210927 TI - Cytopathogenicity of pestiviruses isolated post mortem from cattle. AB - Cytopathic pestivirus was isolated from different tissues of only eight of 23 cattle with mucosal disease. Three persistently infected cows were healthy until slaughter after death of all their seven offspring, out of which one of four examined demonstrated cytopathic pestivirus. PMID- 9210928 TI - Diaplacental infections with ruminant pestiviruses. AB - Pestiviruses are capable of causing diaplacental infections. Maternal viremias are important for localizing virus in the ruminant placentome. Placental lesions occur with cytopathic BVDV and noncytopathic BDV. The ruminant fetus is very susceptible to pestivirus infections once the virus crosses the placenta because the fetus is 1) agammaglobulinemic, 2) immunologically immature, and 3) it has many immature organ systems with undifferentiated cells. Cytopathic BVDV (NADL) in calves and noncytopathic BDV (BD-31) in lambs cause a variety of clinical syndromes including early embryonic death, abortion, stillbirth, malformed fetuses, and/or low birth weight with viral persistence and immunological tolerance. The cytopathic BVDV (NADL) reviewed herein caused pulmonary, placental and dermal lesions when infection occurred at 80-90 days gestation. In contrast, infection at 140-150 days resulted in retinal dysplasia and cerebellar hypoplasia. The lesions were attributed to direct viral cytopathology. Noncytopathic BDV (BD-31) in lambs caused weak lambs, with hairy fleece and tonic clonic tremors. The lambs were of low birth weight, persistently viremic and immunologically tolerant. The lambs are hypothyroid and had severe hypomyelination. It is hypothesized that the central lesion leading to many of the neural, skeletal and dermal lesions was the endocrine dysfunction leading to hypothyroidism. PMID- 9210930 TI - Border disease of sheep--aspects for diagnostic and epidemiologic consideration. AB - Border Disease (BD) is a condition of newborn sheep that results from congenital infection by a non-cytopathic pestivirus, occurring during the first half of gestation. The variations in expression of the virus directly relate to the age of the fetus at the time of infection. There are four distinct disease syndromes: (1) early embryonic death, (2) abortion and stillbirth, (3) birth of lambs with malformations, and (4) birth of small, weak lambs, lacking characteristic clinical signs, but bearing features of immunosuppression. In the newborn, the BD virus may be recovered from all tissues and teratogenic lesions are found in the endocrine, nervous, skeletal, integumentary and immune systems. These effects of virus infection are manifest in the clinical signs characteristic of the disease, such as tremors, ataxia, hairy birthcoat, low birth weight, facial bone malformations, short-boxy stature, and eye abnormalities. The consequences of the BD compromised immune system is an increased susceptibility to infection, a failure to produce specific antibody to BD virus, and an inability to clear the virus; features characteristic of the immuno-tolerant state. The lifelong shedding and persistence of virus is of epidemiologic importance. The persistently infected BD ewe remains a source of infection for the flock both through horizontal transmission (virus shedding) and congenital transmission (a persistently infected ewe will always bear a BD lamb). Detection of persistently infected individuals within a flock is difficult: clinical signs abate with time and most frequently no antibody to BD is produced. PMID- 9210929 TI - The pathways for bovine virus diarrhoea virus biotypes in the pathogenesis of disease. AB - BVDV infections of cattle ranges from the transient acute infections, which may be inapparent or mild, to mucosal disease which is inevitably fatal. On occasions the acute infections can lead to clinical episodes of diarrhoea an agalactia but as these syndromes cannot be reproduced experimentally, the pathogenesis remains unclear. The immunosuppressive effect of acute BVDV infections can enhance the clinical disease of other pathogens and this may be an important part of the calf respiratory disease complex. Although BVDV antigen has been demonstrated within the lymphoid tissues, for prolonged periods, the evidence for viral latency remains to be proven. Venereal infection is shown to be important in the transfer of virus to the foetus and congenital infections can cause abortions, malformations and the development of persistently viraemic calves. The two biotypes of BVDV, non-cytopathogenic and cytopathogenic, are described. Their sequential role in the pathogenesis of mucosal disease arises from the initial foetal infection with the non-cytopathogenic virus and the subsequent production of persistently viraemic calves. These calves may later develop mucosal disease as a result of superinfection with a "homologous" cytopathogenic virus and the possible origin of this biotype by mutation is discussed. Chronic disease is defined as a progressive wasting and usually diarrhoeic condition; it is suggested that this may develop following superinfection of persistently viraemic cattle with a "heterologous" cytopathogenic biotype. PMID- 9210931 TI - A study of some pathogenetic aspects of bovine viral diarrhea virus infection. AB - The cytopathic (CP) strain TVM-2 of bovine virus diarrhea virus (BVDV) induced in calves a severe disease, whereas the calves inoculated with the non-cytopathic (NCP) New York-1 strain, remained clinically normal. When calves were immunosuppressed with dexamethasone (DMS) they underwent an overt, generally fatal disease. This result was obtained with either the CP and the NCP strain of BVDV. It was speculated that the immunosuppressive activity of BVDV could be a property peculiar to certain isolates of the virus. PMID- 9210932 TI - Distribution of antigen of noncytopathogenic and cytopathogenic bovine virus diarrhea virus biotypes in the intestinal tract of calves following experimental production of mucosal disease. AB - Mucosal disease can be experimentally induced by inoculating calves persistently viremic with noncytopathogenic (ncp) Bovine Virus Diarrhea Virus (BVDV) with an antigenetically closely related cytopathogenic (cp) BVDV strain. Calves suffering from mucosal disease develop severe intestinal lesions causing breakdown of the gastrointestinal barrier and death. Knowledge about tissue distribution of ncp/cp biotypes of BVDV may contribute to the understanding of the pathogenesis of these lesions. Distribution of cpBVDV versus ncpBVDV was demonstrated in the intestinal tract of nine calves with experimentally induced mucosal disease and in five persistently viremic calves. Biotypes were distinguished immunohistochemically in organ tissues using monoclonal antibodies against marker epitopes on the viral surface glycoprotein gp53. In persistently viremic calves ncpBVDV was present in a few epithelial cells, mononuclear cells and intramural ganglia. A multifocal reaction was observed in vascular walls. In calves with mucosal disease a striking increase of antigen containing cells occurred. Viral antigen in these cells reacted with marker antibodies for cpBVDV. A distinct tissue distribution of biotypes was observed in intramural ganglia and duodenal glands. Severe tissue damage was correlated to the presence of cpBVDV antigen. This indicates the importance of cpBVDV for the development of lesions. Interactions of cpBVDV and immunemediated mechanisms will need further investigation. PMID- 9210933 TI - Clinical and virological observations of a mucosal disease outbreak with persistently-infected seropositive survivors. AB - A group of 14 four to nine month old calves, clinically healthy but persistently infected with bovine virus diarrhoea virus (BVDV), was obtained from a single farm, and reared as a group. Ten of them were male and were castrated soon after arrival. Signs of mucosal disease (MD) developed within a month and eight of the males had died or been killed on humane grounds by 2 months after purchase. The other two males and one of the females developed more chronic but progressive signs of MD and were killed during the next four months. The remaining three females showed only transient signs of MD followed by clinical recovery. They subsequently remained healthy up to slaughter at 2, 2.5 and 5 years respectively. These three survivors were persistently infected with BVDV, and shed virus in their mucous secretions, although two of them were also seropositive to the virus with fluctuating neutralizing antibody titres (at times as high as 1/960) to a range of BVDV strains including their own persisting virus. PMID- 9210934 TI - Insertion of cellular sequences in the genome of bovine viral diarrhea virus. AB - The genomic sequences of four pestiviruses, two BVDV strains (Osloss and NADL, both of which are cytopathogenic) and two HCV strains, were analyzed. Comparative studies revealed the presence of small insertions of cellular sequences in the genomes of both BVDV strains; the insertions are located in a region coding for a nonstructural protein. Such insertions are not present in the HCV sequences. The insertion identified in BVDV Osloss encodes a complete ubiquitin-like element. The sequence inserted in the BVDV NADL genome shows no homology to a ubiquitin gene but is almost identical with another bovine mRNA sequence. Molecular characterization of a BVDV "pair", isolated from an animal with mucosal disease, led to the detection of a ubiquitin-like sequence in the genome of the cytopathogenic strain, but not of the noncytopathogenic strain. It is proposed that recombination between viral and cellular RNA leads to formation of cpBVDV genomes. This hypothesis has direct implications for the pathogenesis of mucosal disease. PMID- 9210935 TI - Congenital curly haircoat as a symptom of persistent infection with bovine virus diarrhoea virus in calves. AB - Ten calves were born small and with a curly haircoat in a dairy herd which comprised approximately 185 milking animals. These calves commonly developed diarrhoea and/or signs of respiratory disease at the age of 2 to 4 weeks. Two of the calves died and 5 were chronically ill and poor doers and were therefore euthanized. This susceptibility to disease of the curly haired calves was quite different from what was observed among other calves in the herd. Sera from seven of the curly haired calves were examined and were all found to be free from detectable antibodies to bovine virus diarrhoea virus (BVDV) and to harbour a non cytopathic strain of BVDV. One of the calves was retested after 7 weeks and was still seronegative and viraemic. Of 49 non-curly haired calves examined in the herd 44 were BVDV seropositive. The other 5 were seronegative to BVDV but attempts to isolate BVDV from their sera failed. PMID- 9210936 TI - Identification and production of pestivirus proteins for diagnostic and vaccination purposes. AB - Using a panel of monoclonal antibodies (MAbs) previously characterized by seroneutralization, immunofluorescence and radioimmunoprecipitation, we have identified Pestivirus proteins useful for diagnostic purposes from the cytopathic Osloss isolate of bovine viral diarrhea virus (BVDV). Proteins that should be useful for vaccination have also been analysed. Cell-free translation of RNA from glycoprotein-coding cDNA fragments produced, when synthesized in the presence of canine pancreatic microsomes, two glycosylated proteins that were independently recognized and immunoprecipitated by two distinct classes of neutralizing MAbs. A similar in vitro procedure was carried out on nonstructural protein-coding sequences and allowed to identify a viral translation product that specifically reacted with MAbs directed against the 80 kDA protein of a number of Pestivirus strains. Its positioning within the polyprotein encoded by the viral genome was refined by epitope scanning using synthetic hexameric peptides. This viral antigen was further expressed in E. coli, produced as inclusion bodies and used successfully as an ELISA antigen in both competitive and indirect assays for the detection of BVD antibodies in cattle sera. PMID- 9210937 TI - Surveillance of cattle herds for bovine virus diarrhoea virus (BVDV)-infection using data on reproduction and calf mortality. AB - The effect of bovine virus diarrhoea virus (BVDV)-infection on pregnancy rate, on stillbirths and mortality of neonatal calves and the size of newborn calves was evaluated in 8 herds in which persistently infected (PI)-animals had been identified. Data from 9 herds without PI-animals were used as controls. At the time of conception of the oldest PI-animal a significant drop in pregnancy rate to about half the herd average was found. About 6 months later a 3-fold rise in calf mortality was seen. This pattern was found in 4 of the herds. In the remaining 4 herds the pattern was less clear, probably reflecting different immune states of the herds. Rough estimation of the size of newborn calves showed that PI-animals were significantly smaller than normal animals. Monitoring of herds for the above-mentioned parameters may be a means of pointing out herds with PI-animals. Most of the data necessary for such surveillance schemes are already available and may readily be used. PMID- 9210938 TI - Identification of cattle infected with bovine virus diarrhoea virus using a monoclonal antibody capture ELISA. AB - A monoclonal antibody capture enzyme linked immunosorbent assay (ELISA) has been developed to detect pestivirus-specific antigen in the leucocytes of cattle infected with bovine virus diarrhoea virus (BVDV). A blind trial was conducted to compare the specificity of the ELISA with conventional tissue culture virus isolation on 215 blood samples submitted for BVDV diagnosis from cattle throughout Scotland. One hundred and sixty seven samples were negative by both ELISA and virus isolation and 47 samples were positive by both tests. One blood was negative by ELISA and positive by virus isolation. PMID- 9210939 TI - Detection of border disease virus in sheep efferent lymphocytes by immunocytochemical and in situ hybridisation techniques. AB - The prefemoral efferent lymphatics of four sheep persistently infected with a non cytopathic (NCP) isolate of border disease virus (BDV) were cannulated. Recovered lymphocytes were examined for the presence of virus by an immunocytochemical technique employing a pool of monoclonal antibodies which recognise the 120K non structural polypeptide of NCP BDV. The results revealed that 9.5% of the lymphocytes carried virus antigen. Lymphocytes from two of the sheep were studied by in situ hybridisation using a viral antisense RNA probe complementary to the region of the BDV genome coding for the 120K polypeptide. This showed that 70-80% of the cells were infected, confirming the greater sensitivity of the in situ hybridisation technique. PMID- 9210940 TI - Bovine viral diarrhea virus infection: rapid diagnosis by the polymerase chain reaction. AB - The polymerase chain reaction (PCR) was applied to detect bovine viral diarrhea virus (BVDV) by amplification of its nucleic acid sequences in cell cultures, in serum samples of persistently infected cattle, and in organ specimens of acutely diseased calves. The primers and the probes were selected from the gp48 region of the cytopathic NADL strain. The products of single PCR or double PCR were identified by electrophoresis as well as by hybridization with biotinylated probes. The results thus obtained correlated with those of conventional diagnostic procedures, i.e., virus isolation and serology. The detection assay of the BVDV genome by the PCR amplification proved to be both specific and sensitive. PMID- 9210941 TI - cDNA probes for the detection of pestiviruses. AB - Probes were prepared from genomic RNA of Hog Cholera Virus (HCV) after synthesis of cDNA and cloning. Six probes were selected according to their place on the viral genome determined by sequencing and comparison with BVDV sequence. These probes were hybridized with two strains of HCV (Alfort and Nord), two strains of Bovine Viral Diarrhea (BVDV) (NADL, New York) and four strains of Border Disease (BD) (Lyon 1, Lyon 2, Aveyron, IEMVT). This panel of six probes seem to be able to differentiate pestiviruses but some differences rely only on slight intensity of the hybridization. PMID- 9210942 TI - Detection of persistent bovine viral diarrhea virus infections by DNA hybridization and polymerase chain reaction assay. AB - The detection of persistent bovine viral diarrhea virus (BVDV) infections in cattle by DNA dot blot hybridization was done with a cloned cDNA probe prepared from noncytopathic BVDV (strain NY-1). Due to the variability of specific hybridization results, detection of BVDV by primer-directed polymerase chain amplification was done. Primers were chosen within a reported area of sequence conservation and amino acid homology of Pestiviruses. The amplification region extended from nucleotide 6322 to 7475 and was based on published BVDV sequence data (NADL strain). BVDV RNA was extracted by two methods (proteinase K and guanidinium isothiocyanate) from serum and white blood cell preparations collected from 3 persistently-infected heifers. cDNA was synthesized from extracted BVDV-genomic RNA using reverse transcriptase. Reaction conditions were optimized to amplify the 1153 base pair fragment from the cDNA preparation. Detection of BVDV in the samples by DNA dot blot hybridization using a nucleic acid probe corresponding to the amplified region (6322 to 7475) was compared with polymerase chain reaction assay. The increased sensitivity of the polymerase chain reaction assay provided clearer identification of persistently-infected animals than DNA hybridization under similar conditions. PMID- 9210943 TI - Differentiation of pestiviruses by a hog cholera virus-specific genetic probe. AB - A hog cholera virus (HCV)-specific genetic probe has been generated after cloning of the genomic viral RNA. This probe distinguished between HCV and the closely related bovine viral diarrhoea virus (BVDV). Furthermore, it detected a broad spectrum of HCV strains and isolates which differ in their phenotype such as virulence. PMID- 9210944 TI - Lesions in aborted bovine fetuses and placenta associated with bovine viral diarrhoea virus infection. AB - Abortions in dairy cattle were investigated on 55 dairy farms sited in North West England, using a multi-level diagnostic technique. After pathological examination of fetal and placental tissues collected at the time of abortion, possible causes for these abortions could be identified, supported by bacteriological and serological laboratory findings. Of 150 abortions investigated, Bovine Viral Diarrhoea (BVD) virus infection was related to 40 episodes (27% of the total), often accompanied by evidence of concurrent infections. Lesions associated with BVD abortions were found in fetal eyelid, lung, and occasionally myocardium. Lesions in the lung were most consistent, characterized by mononuclear inflammatory cell infiltration of peribronchiolar and inter-alveolar tissues. Placental lesions were non-specific. It is concluded that the lesions observed are insufficient to be the primary cause of abortion. However, the pathological changes associated with BVD infection in the placenta may allow secondary opportunist pathogens to cross the feto-maternal barrier, thereby threatening the health of the fetus and the physiological and endocrinological functions of the placenta which maintain pregnancy. PMID- 9210945 TI - Immunological reactivity of bovine viral diarrhea virus proteins after proteolytic treatment. AB - The immunological reactivity of bovine viral diarrhea virus proteins after proteolytic treatment is described. The results indicate that the epitopes detected are very dependent on conformation of the protein. A partially protease resistant 22 kD fragment of the biotype-specific p80 is identified. PMID- 9210946 TI - Polymerase chain reaction amplification of segments of pestivirus genomes. AB - Reverse transcription followed by polymerase chain reaction (PCR) amplification of a region of the viral genome at the 3' end of the glycoprotein(s) gene was employed with the aim of determining its applicability as a diagnostic tool for pestiviruses. Candidate primers were designed, from homologous segments detected by comparison between the sequences of strains NADL, Osloss and Alfort. A segment of 634 base pairs on the glycoprotein gene was targeted for amplification. Segments of five pestivirus strains of bovine viral diarrhoea virus, two of border disease virus and the Alfort 187 strain of hog cholera virus were amplified successfully. PMID- 9210947 TI - Production of monoclonal antibodies to study the molecular biology of bovine viral diarrhea virus. AB - Five monoclonal antibodies produced against bovine viral diarrhea virus were characterized for some of their biological activities. All of them bound to varying degrees to pestivirus strains but failed to neutralize the virus. One of the antibodies immunoprecipitated four polypeptides presumably involved in viral envelope organization. PMID- 9210948 TI - Determination of level of antibodies to bovine virus diarrhoea virus (BVDV) in bulk tank milk as a tool in the diagnosis and prophylaxis of BVDV infections in dairy herds. AB - An indirect ELISA has been evaluated for determination of the level of antibodies to BVDV in individual milk samples and recently in bulk tank milk from dairy herds. As part of an epidemiological study, bulk milk and individual milk samples from all cows in 15 dairy herds were analysed for antibodies to BVDV two times one year apart. There was an excellent correlation between the level of antibodies in the bulk tank milk and the prevalence of BVDV antibody positive cows. The mean prevalence of BVDV antibody positive cows in the 15 dairy herds was 45.5% (188/413) at the first sampling and 46.2% (191/413) one year later. Seven of the herds had no, or only a low number of antibody positive cows. In contrast, between 52 to 100% of the cows in seven other herds were antibody positive to BVDV. In the 15th herd all cows without antibodies at the first sampling were antibody positive to BVDV one year later, indicating a recently introduced BVDV infection in this herd. Analysis of bulk milk samples for BVDV antibodies is now routinely used in Sweden as a tool in diagnosis and prophylaxis of BVDV infections in dairy herds. The importance and advantages of this diagnostic technique, that has made it possible to establish BVDV-free dairy herds, is discussed. PMID- 9210949 TI - BVD-virus infection in goats-experimental studies on transplacental transmissibility of the virus and its effect on reproduction. AB - Eight groups of altogether 25 goats without neutralizing antibodies against BVD virus, were inoculated either intranasally or intranasally and subcutaneously with two different BVD virus isolates during different stages of gestation. In all 18 goats inoculated within the first 78 days of gestation an abortion and foetal death rate of approximately 100% occurred. Only one goat gave birth to a clinically healthy kid. The other seven goats which were inoculated after the 78th day of gestation showed also a high foetal death rate. Only two of them gave birth to clinically healthy kids. Neutralizing antibodies against BVD virus could be detected in blood samples drawn from 14 kids born at normal term including stillborn and non-viable offsprings. BVD virus was reisolated from different organs taken from seven foetuses. It was not possible to isolate BVD virus from any of the normal offsprings. PMID- 9210950 TI - BVD virus isolation techniques for routine use in cattle herds with or without previous BVD history. AB - Buffy coats of 1074 cattle were tested for BVD virus using the usual longterm cultivation (LTC) in bovine kidney monolayer cell cultures (7 days) whereby 268 BVD virus carriers could be detected. Serum samples collected simultaneously from the same animals were examined by means of a shortterm-cultivation (STC) procedure of only two days in stationary macroplate cell cultures. Using this method only 172 amongst the former 268 BVD virus carriers were found. Of the remaining 96 serum samples from animals positive in buffy coats leucocytes by LTC and negative in sera by STC, further 19 cattle were found to be viraemic when the sera were additionally tested by LTC. These results are discussed with regard to the antibody level and the age of the animals. The reduced sensitivity of STC of sera is considered in relation to the favourable time and cost factor. STC of serum samples in connection with the serological results on a herd basis proved to be valuable for the examination of cattle of more than 6 months of age but not for calves below 6 months. This was particularly true in cattle herds with no previous BVD history. PMID- 9210951 TI - Molecular characterisation of the coding region for the p125 from homologous BVDV biotypes. AB - We amplified and sequenced the p125 coding regions of a 'homologous' pair of BVDV biotypes, Pe515 cytopathogenic and non-cytopathogenic. The sequences were aligned with the published sequences of Osloss, NADL and the HCV Alfort strains, but no insertions of host sequence were observed in that region. PMID- 9210952 TI - Progeny of sheep persistently infected with border disease virus. AB - Most lambs affected with border disease die early in life but those which survive gradually loose their body tremors and their fleece abnormalities become less clear. Seven female lambs persistently infected with border disease virus were reared to maturity and bred from when they were 2 to 3 years old. Two failed to conceive but five gave birth to 6 live lambs with clinical signs of border disease characterized by hairy and pigmented fleece with or without body tremors. The epidemiological significance of persistently infected sheep is discussed. PMID- 9210953 TI - A deletion polymorphism due to Alu-Alu recombination in intron 2 of the retinoblastoma gene: association with human gliomas. AB - The retinoblastoma gene (RB) encodes a tumor suppressor that is inactivated in a number of different types of cancer. We searched for gross alterations of this gene in tumors of the central nervous system by using Southern blot hybridization. A common alteration was found in several tumors and was mapped to the region around exon 2. Nucleotide sequencing showed that the alteration was caused by a 799-bp deletion in intron 2 of the RB gene and was probably due to homologous recombination between two Alu repeats. Deletions of this type have not been found previously in the RB gene. The deletion turned out to be a polymorphism with an allele frequency estimated at 2.2% in 185 patients without cancer. The deletion was found in five of 48 patients with brain tumors (allele frequency of 5.2%). This difference is not statistically significant (P = 0.149, Fisher's exact test). Confining the analysis only to glioma brain tumors revealed a statistically significant difference compared with the cancer-free patient controls (P = 0.027, Fisher's exact test). Further study is needed to determine if the deletion is a weak brain cancer-predisposing mutation or a harmless polymorphism. Finding this mutation in a tumor and the germline DNA of a retinoblastoma patient could lead to incorrect estimation of the heritability of a tumor. PMID- 9210954 TI - Induction of p21/WAF1 and G1 cell-cycle arrest by the chemopreventive agent apigenin. AB - Apigenin is a plant flavonoid that has been shown to significantly inhibit ultraviolet-induced mouse skin tumorigenesis when applied topically and may be an alternative sunscreen agent for humans. A long-term goal of our laboratory is to elucidate the molecular mechanism or mechanism by which apigenin inhibits skin tumorigenesis. In a previous publication, we characterized the mechanism by which apigenin induced G2/M arrest in keratinocytes. More recent studies in our laboratory have provided evidence that apigenin can induce G1 arrest in addition to arresting cells at G2/M. Here we describe the mechanism of the apigenin induced G1 arrest in human diploid fibroblasts (HDF). Treatment of asynchronous HDF for 24 h with 10-50 microM apigenin resulted in dose-dependent cell-cycle arrest at both the G0/G1 and G2/M phases as measured by flow cytometry. The G0/G1 arrest was more clearly defined by using HDF that were synchronized in G0 and then released from quiescence by replating at subconfluent densities in medium containing 10-70 microM apigenin. The cells were analyzed for cell-cycle progression or cyclin D1 expression 24 h later. A dose of apigenin as low as 10 microM reduced the percentage of cells in S phase by 20% compared with control cultures treated with solvent alone. Western blot analysis of apigenin-treated HDF indicated that cyclin D1 was expressed at higher levels than in untreated cells, which signifies that they were arrested in G1 phase rather than in a G0 quiescent state. The G1 arrest was further studied by cyclin-dependent kinase 2 (cdk2) immune complex-kinase assays of apigenin-treated asynchronous HDF, which demonstrated a dose-dependent inhibition of cdk2 by apigenin. Inhibition of cdk2 kinase activity in apigenin-treated cells was associated with the accumulation of the hypophosphorylated form of the retinoblastoma (Rb) protein as measured by western blot analysis. The cdk inhibitor p21/WAF1 was also induced in a dose dependent manner, with a 22-fold induction of p21/WAF1 in 70 microM apigenin treated cells. In conclusion, apigenin treatment produced a G1 cell-cycle arrest by inhibiting cdk2 kinase activity and the phosphorylation of Rb and inducing the cdk inhibitor p21/WAF1, all of which may mediate its chemopreventive activities in vivo. To our knowledge this is the first report of a chemopreventive agent inducing p21/WAF1, a known downstream effector of the p53 tumor suppressor protein. PMID- 9210955 TI - Ha-ras oncogene-induced transcription of human papillomavirus type 18 E6 and E7 oncogenes. AB - Human papillomavirus (HPV) DNA sequences are found in most carcinomas originating from the uterine cervix. HPV E6 and E7 oncogenes have been shown to cooperate with ras oncogenes to fully transform human epithelial cells. We investigated the effect of the Ha-ras oncogene on the transcriptional activity of HPV-18 and found that it induced the transcriptional activity of the viral promoter, whereas the normal gene had only a minimal effect. However, transfection of the normal Ha-ras gene and simultaneous inhibition of protein phosphatase sensitive to okadaic acid (OA) resulted in a cooperative transactivation of the viral promoter. When cloned upstream of a minimal promoter, the AP-1 binding sites present in the viral promoter conferred transcriptional responsiveness to Ha-ras and OA. Furthermore, HeLa cell clones permanently expressing the Ha-ras oncogene or high levels of the normal gene exhibited a twofold to threefold increase in E6*E7/E1 and E6*E7 transcripts. We propose that both Ha-ras and a protein phosphatase sensitive to OA regulate HPV oncogene expression through modulation of AP-1 activity and suggest that increased levels of E6 and E7, resulting from activated viral transcription in the presence of ras oncogenes, may in part explain the observed cooperation between these viral and cellular oncogenes in the transformation of human cells. PMID- 9210956 TI - Downregulation of aryl hydrocarbon receptor function and cytochrome P450 1A1 induction by expression of Ha-ras oncogenes. AB - The immortalized human epithelial cell line MCF10A has the phenotypic characteristics of normal breast cells. Exposure of MCF10A cultures to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) stimulated the transcriptional activation of cytochrome P450 1A1 (CYP1A1), and CYP1B1, and NAD(P)H:quinone oxidoreductase. Northern blot hybridization and nuclear run-on assays demonstrated that transcriptional activation of these genes was suppressed in stably transfected cultures expressing an Ha-ras oncogene (the MCF10A-NeoT line). Similar suppression did not occur in stably transfected lines carrying the expression vector or a normal c-Ha-ras protooncogene. Western blot analyses and immunofluorescence microscopy demonstrated that the lack of inducibility in MDF10A-NeoT cells reflected neither reductions in aryl hydrocarbon receptor (AHR) and aryl hydrocarbon nuclear translocator protein nor prevention of TCDD-induced AHR translocation to the nucleus. Suppression did correlate with reductions in DNA-AHR complex formation, as analyzed by gel retardation assays of soluble cell extracts treated in vitro with TCDD. The induction of Cyp1a-1 by TCDD was also analyzed in transgenic mice that expressed a v-Ha-ras oncogene exclusively in their keratinocytes. Relative to littermates lacking the transgene, the induction of Cyp1a-1 by TCDD was partially suppressed (about 50%) in the epidermises of v Ha-ras-positive transgenic mice. However, normal levels of Cyp1a-1 induction occurred in the livers of the same mice. induction of Cyp1a-1 by TCDD was also suppressed (more than 98%) in chemically induced skin papillomas having Ha-ras mutations, relative to uninvolved surrounding skin. These studies suggest that the p21-ras protein controls signal transduction pathways capable of modulating AHR function. PMID- 9210957 TI - Suppression of nitric oxide-induced apoptosis by N-acetyl-L-cysteine through modulation of glutathione, bcl-2, and bax protein levels. AB - It has been demonstrated that nitric oxide (NO) can promote apoptosis in human cancer cells. To test the protective effects of antioxidants (N-acetyl-L-cysteine (LNAC) and free-radical spin traps (5,5-dimethyl-1-pyrroline N-oxide and 2,2,6,6, tetramethyl-1-piperidinyloxy) against NO-induced apoptosis, a human colon cancer cell line (COLO 205) was treated with NO, and its survival rate was evaluated both with and without antioxidant therapy. LNAC arrested the development of progression of apoptosis in COLO 205 cells in a dose-dependent manner, promoted long-term survival, and prevented the internucleosomal DNA cleavage induced by NO. The intracellular level of glutathione (GSH) was found to be elevated in cells after exposure to LNAC. The bax protein levels were elevated by NO treatment, and this effect was blocked by LNAC. On the other hand, the bcl-2 oncoprotein level in the LNAC-pretreated cells was significantly elevated in a time-dependent manner compared to cells that received NO pretreatment. In summary, our results suggest that the protective effect of LNAC may be linked to its inducement of increases in cellular GSH and bcl-2 protein levels and to its suppression of cellular bax protein in treated cells. PMID- 9210958 TI - Complex genomic rearrangement within the 12q15 multiple aberration region induced by integrated human papillomavirus 18 in a cervical carcinoma cell line. AB - Human papillomavirus (HPV) DNA is integrated into the host genome in cervical cancer. The cervical carcinoma cell line SW756 has integrated HPV-18 DNA in chromosome region 12q15, in the papillomavirus-associated locus-2 (PAL2). By polymerase chain reaction and hybridization of an arrayed cosmid library with oligonucleotides from the rearranged allele, we determined the pre-integration germline structure of the region. PAL2 was located approximately 10 kb from sequence-tagged site marker U27131, which was the marker most proximal to the 3' flank of the integrated viral DNA. HPV-18 DNA integration induced a complex genomic rearrangement resulting in inversion and deletion of cellular sequences. PAL2 is within the multiple aberration region, which has been shown to be affected in several types of benign tumors of mesenchymal origin. The integrated viral DNA was located 50 kb from a CpG island and 150 kb upstream of the high mobility group I-C (HMGI-C) gene. The HMGI-C gene and the integrated HPV-18 DNA had opposite transcriptional orientations. No overexpression or altered message of the HMGI-C gene was detected in three cervical carcinoma cell lines. The integrated viral DNA did not affect any other known gene in the region and may be a marker for an unknown gene associated with malignant tumor phenotypes. PMID- 9210959 TI - Opposite effect of stable transfection of bioactive transforming growth factor beta 1 (TGF beta 1) versus exogenous TGF beta 1 treatment on expression of 92-kDa type IV collagenase in mouse skin squamous cell carcinoma CH72 cells. AB - We have previously shown that transforming growth factor-beta 1 (TGF beta 1) mRNA is consistently overexpressed in squamous cell carcinomas relative to normal mouse skin. Here we show that 92-kDa type IV collagenase (matrix metalloproteinase) (MMP-9) mRNA was likewise progressively overexpressed during mouse skin carcinogenesis. To determine if overexpression of MMP-9 and TGF beta 1 are linked, we stably transfected a bioactive TGF beta 1 into a mouse skin squamous cell carcinoma cell line (CH72), which resulted in about twofold to three-fold higher levels of secreted active TGF beta 1. Active TGF beta 1 transfected cells grew only slightly, but not significantly, more slowly in vitro and in vivo than vector-only transfectants. Two clones overexpressing active TGF beta 1 secreted much reduced levels of MMP-9 activity, as determined by zymogram analyses. However, treatment of these clones with 40 pM exogenous TGF beta 1 for 48 h enhanced secretion of MMP-9 activity. Constitutive mRNA expression of MMP-9 was reduced twofold to 70-fold in five untreated active TGF beta 1-transfected clones relative to the other transfectants. In contrast, treatment with 40 pM exogenous TGF beta 1 induced MMP-9 mRNA expression in a time-dependent fashion, from twofold to fourfold after 4 h to a maximum of 12- to 19-fold after 24-48 h. Induction of MMP-9 mRNA was dose dependent at TGF beta 1 concentrations of 4-400 pM. Thus, stable transfection of bioactive TGF beta 1 downregulated whereas exogenous TGF beta 1 treatment upregulated MMP-9 activity and expression. Treatment of transfectants with a neutralizing TGF beta 1 antibody slightly downregulated constitutive MMP-9 mRNA (20-30%) but completely blocked induction by exogenous TGF beta 1. Thus, the effect of TGF beta 1 transfection was not due to secreted TGF beta 1 but may have been a secondary effect. PMID- 9210961 TI - Cataract patients in a defined Swedish population 1986-1990. PMID- 9210960 TI - Assessment of mutations in Ki-ras and p53 in colon cancers from azoxymethane- and dimethylhydrazine-treated rats. AB - Mutations in the Ki-ras oncogene and the p53 tumor suppressor gene are known to occur at high frequencies in human colon cancers. We measured the frequency of mutations in these two genes in colon adenocarcinomas obtained from a widely used experimental model of human colon carcinogenesis: F344 rats treated with the carcinogens azoxymethane (AOM) or dimethylhydrazine (DMH). We detected codon 12 mutations in Ki-ras in approximately 60% of colon adenocarcinomas induced by either carcinogen. We characterized the rat p53 intron-exon junctions to construct primers for polymerase chain reaction amplification of this gene. We discovered that the rat p53 gene was structurally different from the human p53 gene, as the rat gene was missing one intron between exons 6 and 7. Both single stranded DNA conformational polymorphism analysis and direct DNA sequencing of the highly conserved regions of rat exons 5-7 were conducted because the corresponding human regions (exons 5-8) have been reported as being mutated most frequently in human colon cancers. Using these methods, we were unable to identify any p53 mutations in the highly conserved regions of exons 5-7 in either AOM- or DMH-induced colon adenocarcinomas. These data confirm that Ki-ras was mutated in most colon cancers in AOM- or DMH-treated rats but indicate that molecular alterations in the p53 gene, if they occur in this animal model, are different from most p53 mutations in human colon cancers. PMID- 9210962 TI - Prevalence of intestinal parasites in the human population of Leon, Nicaragua. AB - Intestinal parasites appear to be prevalent in Nicaragua, which motivated a more extensive prevalence study in which socioeconomic conditions such as degree of crowding, quality of water supply, type of floor and disposal of excretion, were considered. The study was performed on 1267 stool samples from about 8% of the citizens of the city of Leon. The overall prevalence of intestinal pathogenic parasites among the 1267 individuals was found to be 47.2%. The prevalence of Entamoeba histolytica/dispar was 18.6% followed by Giardia (15.9%) and Ascaris (13.4%). Other helminths such as hookworms and Strongyloides sp. were found at very low rates. Giardia, in contrast to worm infections, was prevalent already in children under 5 years of age. E. histolytica/dispar increased with age and remained high. Of 595 individuals with intestinal parasites 81% were living in 'poor' conditions and in 13 clusters of households, a lower prevalence of parasites was seen in households characterised as having good socioeconomic conditions. However, several variables appear to be important in determining the prevalence of the individual intestinal protozoa and helminths encountered. PMID- 9210963 TI - Ex vivo antimalarial activity of proguanil combined with dapsone against cycloguanil-resistant Plasmodium falciparum isolates. AB - The ex vivo antimalarial activity of plasma samples obtained from 20 healthy Caucasian volunteers following daily proguanil (200 mg) plus dapsone (8 mg) for malaria chemoprophylaxis inhibited five cycloguanil-resistant Thai isolates of Plasmodium falciparum. All volunteers were phenotyped as extensive metabolisers (EMs) of proguanil. Three of the five isolates were obtained from Thai soldiers who had failed malaria prophylaxis on daily proguanil (200 mg) plus dapsone (4.0 or 12.5 mg). The Thai soldiers were also classified as EMs, but had relatively lower plasma cycloguanil concentrations compared to values reported in the literature for Caucasians and black Kenyans. Although the high level of parasite resistance to cycloguanil was the most likely explanation for the Thai soldiers failing prophylaxis on proguanil plus dapsone, their low cycloguanil concentrations may have also contributed to their lack of protection. However, in areas where parasites are more susceptible to cycloguanil, such as in certain regions of Africa, proguanil plus dapsone may still be an effective chemoprophylactic drug combination. PMID- 9210964 TI - In vivo sensitivity of Plasmodium falciparum to chloroquine and sulfadoxine pyrimethamine in school children in Hoima district, western Uganda. AB - In vivo testing of Plasmodium falciparum sensitivity to chloroquine and sulfadoxine-pyrimethamine was carried out among asymptomatic school children from seven schools in four sub-counties in Hoima District in western Uganda. Seven hundred and twenty five children were screened, 487 (67%) had parasitaemia and 307 met the inclusion criteria. Full sensitivity to chloroquine in urban areas was found in 42%. Chloroquine resistance at RII level was observed in 46% (33 58%) and RIII level in 13% (0-24%) In rural areas, however, full sensitivity to chloroquine was found in 76% (70-84%). Resistance at RII and RIII level was found in 18 and 6% respectively. Full sensitivity to sulfadoxine-pyrimethamine was found in 98% and only 2% resistant at RII level. The relatively high proportion of chloroquine resistance at RII and RIII levels in urban areas could be attributed to easy access to chloroquine as well as frequent and probably inadequate use of chloroquine. Chloroquine can still be used as first line antimalarial drug in the area. Sulfadoxine-pyrimethamine should be the second line drug for cases not responding to chloroquine. PMID- 9210965 TI - The use of morphometrics in entomological surveillance of sylvatic foci of Triatoma infestans in Bolivia. AB - Jamach'uma (Cochabamba, Bolivia) is a small village surrounded by sylvatic foci of Triatoma infestans. Houses in the village were also infested with T infestans, and were sprayed in December 1992 as part of a Chagas disease vector control trial. Ten months later the houses were found to be again infested with a few fifth instar nymphs of T. infestans. These nymphs were compared by seven head measurements with 36 fifth instar nymphs collected from houses in Jamach'uma before treatment, and with two sets of nymphs originating from the surrounding sylvatic foci: eight specimens collected in 1992 and nine specimens collected in 1995. The results are discussed in relation to the possible mechanisms of the apparent reinfestation: recrudescence of a residual domestic population or reinvasion of the houses from surrounding sylvatic foci. Quantitative comparisons support the former hypothesis. PMID- 9210966 TI - Plasma chloroquine concentrations in young and older malaria patients treated with chloroquine. AB - Plasma chloroquine (CQ) concentrations were measured by bioassay in young (0-4 years, n = 9) and older (5-60 years, n = 21) patients from Vanuatu infected with malaria following treatment with 25 mg/kg CQ over 3 days. CQ concentrations in young children tended to be lower than in older patients at days 2, 3, 4 and 7 after onset of treatment, with no drug present in two young children on day 3 and in one child on day 7. The greater difficulty experienced by young children to ingest all of their prescribed medication could have contributed to the lower CQ concentrations observed in the younger age group. The possibility that sub therapeutic CQ concentrations are responsible for treatment failures in young children should be considered in areas where a high degree of CQ resistance has not yet been established. In such areas, the presence or prevalence of CQ resistant infections should not be based on treatment failures observed in young children unless it can be confirmed that adequate blood CQ concentrations were achieved after treatment. PMID- 9210967 TI - Cure by ivermectin of a chronic, persistent, intestinal strongyloidosis. AB - A report is given of a cure by ivermectin of a 19-year-old patient with chronic, persistent, 3-year-old intestinal strongyloidosis resistant to several dosage regimens of conventional anthelminthics, including the current drug of choice for strongyloidosis thiabendazole and its therapeutic alternative, albendazole. Ivermectin was administered, first as a single, oral dose of 200 micrograms/kg with consequent reduction in larval output, but no complete parasitological cure. A second course of ivermectin, 200 micrograms/kg administered for two consecutive days, resulted in complete parasitological cure, as evidenced by the absence of Strongyloides stercoralis larvae in stool samples examined through the kato thick smears, Baermanns concentrations as well as in the 'enterotest' performed on jejunal fluid. The patient has remained parasitologically cured, after 7-months follow-up. Ivermectin was well tolerated with mild clinical and biochemical reactions which did not last for long. PMID- 9210968 TI - Countercurrent immunoelectrophoresis test for detection of hydatid antigen in the fluid from hydatid cysts: a preliminary report. AB - The exact aetiology of a suspected hydatid cyst may sometimes be a diagnostic dilemma. One of the recent methods to confirm that the cysts are echinococcal in origin is the demonstration of hydatid antigens in the aspirated cystic fluid. In the present study, we evaluated the use of countercurrent immunoelectrophoresis (CIEP) to detect hydatid antigen in cyst fluids. Antibody used for detecting the antigen consisted of hyper immune sera raised in rabbits after inoculation of crude human hydatid cystic fluid antigen. Fluids were collected post-operatively from a total of 14 hydatid cysts confirmed by surgery and by histopathology. The results of the study show that the test is moderately sensitive detecting antigen in 11 (78.5%) of 14 cyst fluids. The test did not detect antigen in three other cyst fluids. The main advantage is its specificity as it was 100% specific with no reactions in control samples. Even though, sensitivity is not very high, the test is sample, inexpensive, and can rapidly diagnose hydatid aetiology of cysts and may be of help in the diagnosis of hydatid cysts in peripheral parasitology laboratories. PMID- 9210969 TI - Effect of storage on serum vitamin B12 and folate stability. AB - To facilitate transport from remote locations, the stability of vitamin B12 and folate was investigated in serum specimens. Serum vitamin B12 proved to be highly unstable, emphasizing that specimens should be frozen if not analyzed immediately. Light protection is necessary if the sample cannot be analyzed within 4 hours. In contrast, folate is a more robust analyte. In refrigerated serum specimens, folate was stable up to 7 days of storage. In situations where specimen stability is important, vitamin B12 status is better assessed with serum or urine methylmalonic acid measurements. Although folate status can be assessed in a similar fashion with homocysteine, specimen stability indicates that direct measurement of folate is a better strategy. PMID- 9210970 TI - Pathogenic analysis of Aeromonas hydrophila septicemia. AB - Aeromonas hydrophila has emerged as a potential pathogen in the immunocompromised host. Various aeromonal infections, including septicemia, have also been reported in apparently healthy individuals. For years, researchers have disagreed over the epidemiologic roles of aeromonads in gastroenteritis. Isolation rates of aeromonads by stool culture among patients with gastroenteritis are not consistently high. Carriers of this bacterium also exist. The septicemic course is, however, often fulminant and fatal, and may lack an obvious focus. Pathogenic mechanisms are complex and largely unresolved. The objective of this study is to report the necropsy findings from a uremic patient who presented with typical aeromonal septicemia of obscure origin asking if such investigation could give insight into some of the questions mentioned previously. Western blot immunostaining for aerolysin (beta-hemolysin of aeromonads) was used to evaluate whether or not such a virulence factor is involved in the process of septic dissemination. The autopsy showed that the skin and liver contained microabscesses. The upper gastrointestinal mucosae and spleen contain patchy putrefactive lesions with adjacent focal hemorrhage. Perimortem blood cultures grew Aeromonas hydrophila. A conventional Western blot analysis of the culture supernatant failed to show aerolysin. A control Aeromonas sobia American Type Culture Collection (ATCC) strain produces readily detectable aerolysin. It is concluded that this isolate may be aerolysin-deficient or one secreting low levels of aerolysin; these would require more sensitive methods of detection. The primary focus of infection might be the upper gastrointestinal tract. Other virulence factors including the bacterial proteases and/or phospholipases might be responsible for the pathogenesis of septic dissemination. PMID- 9210971 TI - Hoechst 33342-induced apoptosis in BC3H-1 myocytes. AB - Bisbenzimidazoles (Hoechst 33342 and Hoechst 33258) are cell permeable, adenine thymine binding fluorescent dyes used to stain deoxyribonucleic acid (DNA) during the evaluation of cell cycle, induction of apoptosis by various ligands and cell viability by flow cytometry. These dyes inhibit topoisomerase I activity in vitro, like camptothecin. In this study, Hoechst 33342 is shown to induce apoptosis at concentration of 10 micrograms/mL or greater after 3 hours incubation in Dulbecco's Modified Eagle Medium characterized by rounded cell morphology, half-moon nuclei with condensed chromatin and a DNA fragmentation ladder of 180 base pair multiples. Hoechst 33258 at the same molarity or seven times greater molarity did not induce apoptosis. If the BC3H-1 myocytes were incubated in RPMI-1640 media, two times the concentration of Hoechst 33342 (20 micrograms/mL) was required to initiate apoptosis. Staining of unfixed cells with Hoechst 33342 may induce apoptosis in the absence of ligands. Therefore, Hoechst 33342 concentration and staining interval should be tested before ligands which may induce apoptosis are evaluated. PMID- 9210972 TI - Further characterization of a monoclonal antibody recognizing apolipoprotein E peptides in amyloid deposits. AB - A monoclonal antibody (YK-2), which was previously established to react with apolipoprotein E (apoE) peptides in systemic amyloid deposits, was further characterized. Epitope of this antibody was determined to be the residue 221 to 230 of apoE. In comparison with polyclonal anti-apoE antibodies, this antibody showed strong reactivity with apoE peptides in amyloid fibril preparation but poor reactivity with native apoE protein or apoE in serum, indicating its usefulness for probing degraded apoE in amyloid deposits. Immunohistochemical studies resulted in strong reactivity for amyloid A and immunoglobulin light chain deposits but weak for beta 2-microglobulin and beta amyloid (senile plaque) deposits. Although the association of apoE with amyloid is non-specific to the component peptide of amyloid fibrils, present findings suggest that the amount or degradation manner of apoE or the environment around the antibody epitope vary among types of amyloidosis. PMID- 9210973 TI - Inhibitory effects of seven organosulphur compounds on clinical isolates of Candida species in vitro. AB - Thirty clinical isolates of Candida albicans and 10 other Candida species were tested for susceptibility to 6 substituted dithiocarbamates and one dimercaptosuccinate. Dimethyldithiocarbamate, sodium pyrrolidine dithiocarbamate, and sodium diethyldithiocarbamate showed dose-dependent antifungal activity which was partially reversed by the addition of zinc, copper, or iron sulfate with greatest reversal at 2:1 metal to dithiocarbamate molar ratio. Anaerobiosis also interfered with dithiocarbamate antifungal activity. PMID- 9210974 TI - Release of amino acids from fatty livers during organ harvest for transplantation. AB - Shortage of organ donors presents a perplexing problem in liver transplantation, and improved methods for evaluating the viability of organs prior to implantation are urgently needed. In the present study, the hypothesis was evaluated that grafts from fatty livers release more amino acids than non-fatty controls during organ harvest. Amino acids in graft rinse effluents at the time of harvest and after cold storage were measured by reverse-phase high performance liquid chromatography and compared with plasma aspartate aminotransferase (AST) levels and recipient survival. Twenty-four hours after transplantation of fatty livers, AST levels in recipient rats were increased more than two-fold compared to non fatty controls (p < 0.01). Survival in the control group was 83 percent, whereas animals receiving fatty livers from ethanol-treated rats survived no longer than 7 days after transplantation (p < 0.05). The rate of release of amino acids from the liver explant was two-fold higher during the harvest procedure (0.5 h) than during the subsequent 23.5 hour cold storage period (435 +/- 70 vs. 186 +/- 14 nmol/ml/hr/g liver, p < 0.001). Further, in the early rinse effluent, amino acids were released about two-fold faster from fatty livers than from controls (p < 0.05). This study demonstrates that the release of amino acids from liver explants increases during the harvesting procedure and is about two-fold higher in fatty livers which fail after transplantation than in surviving controls. It is proposed that amino acid release from explants after organ harvest might serve as a useful marker to evaluate graft function prior to transplantation. PMID- 9210975 TI - Disappearing trisomy 8 mosaicism. AB - A case is presented of a patient with disappearing trisomy 8 mosaicism initially thought to have stigmata of the fragile X syndrome. This case is interesting for two reasons. First, it demonstrates the occurrence of "disappearing mosaicism," a phenomenon first described by LaMarche et al, in 1967. Our patient, initially studied in 1991 by two laboratories and found to be mosaic for chromosome 8 trisomy, was apparently normal by both GTG-banding and fluorescent in situ hybridization (FISH) when studied in 1996. Second, this case further underscores the fact that except under special circumstances, it is important that GTG banding analysis be performed so that the entire human genome be examined in addition to scoring for the fragile X mutation on Xq27.3. In a recent review of the existing database at Rhode Island Hospital on chromosomal abnormalities found in patients referred because of a question of the fragile X syndrome during the period from January 1, 1990 to June 30, 1995, it was found that the frequency of other chromosomal abnormalities in patients referred because of a question of fragile X syndrome equaled or exceeded that of patients found to be positive for fragile X. Our figures, consistent with those reported in the literature, underscore the value of routine karyotyping in this population of patients. PMID- 9210976 TI - Effect of phenobarbital, 3-methylcholanthrene, and chloramphenicol pretreatment on the pharmacokinetics and pharmacodynamics of azosemide in rats. AB - The effects of pretreatment with the enzyme inducers phenobarbital (PB) and 3 methylcholanthrene (3-MC) and the enzyme inhibitor chloramphenicol (CM) on the pharmacokinetic and pharmacodynamic parameters of azosemide were examined after intravenous (i.v.) administration of azosemide, 10 mg kg-1, to rats. The nonrenal clearance (1.63 versus 3.30 mL min-1 kg-1) of azosemide increased significantly in 3-MC pretreated rats. This suggested that the nonrenal metabolism of azosemide increased by pretreatment with 3-MC. This relationship was supported by the significant decrease in 24 h urinary excretion of unchanged azosemide in 3-MC pretreated rats (54.1 versus 41.1% of i.v. dose). This relationship was also supported at least in part by the results of a liver homogenate study; the amount of azosemide remaining per gram of liver decreased significantly (48.2 versus 43.0 micrograms) and the amount of M1 formed increased significantly (4.88 versus 6.66 micrograms when expressed in terms of azosemide) in 3-MC pretreated rats after 30 min incubation of 50 micrograms azosemide in 9000 g supernatant fractions of liver homogenates. The content of hepatic cytochrome P-450 (0.751 versus 1.57 nmol/mg protein) and the weight of liver (3.53 versus 4.20% of body weight) increased significantly in 3-MC pretreated rats, suggesting that the metabolizing enzyme(s) for azosemide seemed to be induced by pretreatment with 3 MC. The 8 h urine output (29.2 versus 18.1 mL) and 8 h urinary excretion of sodium (4.02 versus 2.39 mmol) and chloride (4.01 versus 2.73 mmol) per 100 g body weight decreased significantly in 3-MC pretreated rats. However, the diuretic, natriuretic, kaluretic, and chloruretic efficiencies were not significantly different between the control and 3-MC pretreated rats. The pharmacokinetic and pharmacodynamic parameters of azosemide were not significantly different between the control and PB pretreated rats, and similar results were also obtained from the control and CM pretreated rats. The above data indicate that the metabolizing enzyme(s) for azosemide seem(s) to be neither induced by PB pretreatment nor inhibited by CM pretreatment. However, the content of hepatic cytochrome P-450 and the weight of liver increased significantly in PB pretreated rats, while the values were not significantly different between the control and CM pretreated rats. PMID- 9210977 TI - Pharmacokinetics of the enantiomers of verapamil after intravenous and oral administration of racemic verapamil in a rat model. AB - Verapamil is a chiral calcium channel blocking drug which is useful clinically as the racemate in treating hypertension and arrhythmia. The published pharmacokinetic data for verapamil enantiomers in the rat model are limited. Utilizing a stereospecific high-performance liquid chromatographic (HPLC) assay, the enantiomeric disposition of verapamil is reported after intravenous (1.0 mg kg-1) and oral (10 mg kg-1) administration of racemic verapamil to the rat model. After intravenous administration the systemic clearance of R-verapamil was significantly greater than that of S-verapamil; 34.9 +/- 7 against 23.7 +/- 3.7 mL min-1 kg-1 (mean +/- SD), respectively. After oral administration, the clearance of R-verapamil was significantly greater than that of S-verapamil, 889 +/- 294 against 351 +/- 109 mL min-1 kg-1, respectively. The apparent oral bioavailability of S-verapamil was greater than that of R-verapamil, 0.074 +/- 0.031 against 0.041 +/- 0.011, respectively. These data suggest that the disposition of verapamil in the rat is stereoselective; verapamil undergoes extensive stereoselective first-pass clearance after oral administration and the direction of stereoselectivity in plasma is opposite to that observed in the human. PMID- 9210978 TI - Pharmacokinetics of acebutolol enantiomers after intravenous administration of racemate in a rat model: a dosing range comparison. AB - Acebutolol (AC) is a chiral beta-adrenergic blocking drug, possessing intrinsic sympathomimetic activity (ISA), and is useful clinically as the racemate in treating hypertension. Utilizing a stereospecific high-performance liquid chromatographic (HPLC) assay, the enantiomeric disposition of AC and its major metabolite diacetolol (DC) are reported after intravenous administration of single 5, 15, 30, and 50 mg kg-1 doses of racemate to male Sprague-Dawley rats. The mean area under the plasma concentration versus time curve (AUC) values display a linear relationship with respect to the administered dose. No statistical differences are observed in apparent volume of distribution (Vd), terminal elimination half-life (t1/2), total body clearance (Clt), or renal clearance (Clr) with respect to dose administered. Generally, R-S ratios for AUC following AC administration are statistically different from unity (p < 0.05). However, for the 50 mg kg-1 doses the R-S ratio for AUC is not statistically different from one. For DC, the plasma disposition is nonstereoselective in plasma. The amount of R-DC recovered in urine, however, was greater than that of the antipode (R:S = 1.92 +/- 0.29). This study suggests that the enantiomeric disposition of intravenous AC is linear within the investigated range of 5-50 mg kg-1 racemate in rats. PMID- 9210979 TI - Percutaneous absorption and disposition studies of methotrexate in rabbits and rats. AB - The absorption and disposition of methotrexate (MTX) in the plasma, synovial fluid (SF), skin, and muscle tissue were studied following administration of a topical MTX gel in rabbits and rats. In rabbits, MTX concentrations in the plasma increased steadily toward the peak (5.9 +/- 2.8 ng mL-1) which appeared at approximately 2 h postdose and declined with the elimination half-life of 4.48 +/ 1.74 h. At 1 h after the topical dose, the MTX concentrations in the skin (49.0 +/- 19.8 micrograms g-1), muscle (12.7 +/- 3.3 ng g-1), and SF (19.2 +/- 10.1 ng g-1) underneath the dosed stifle joint were significantly higher (p < 0.05) than those of the untreated stifle joint, indicating the potential therapeutic value of topical delivery of MTX for rheumatoid arthritis. A large fraction (approximately 59%) of MTX which was found in the skin at 1 h postdose was present in the stratum corneum, indicating its extensive binding capacity for MTX. The MTX concentrations in the muscle and SF of the dosed stifle joint at 1 h postdose were 1.8 and 2.6 times higher than those in the dosed elbow joint, respectively, reflecting the effect of dose site on the permeation of MTX. Using a new filter paper method, the amounts of SF obtained from the elbow and stifle joints of four rabbits were 26.3 +/- 8.3 and 48.8 +/- 5.2 mg, respectively. A significant enhancer effect of N,N-diethyl-n-toluamide (DEET) on the disposition of MTX in the stratum corneum of rabbit ear was observed (p < 0.05) by the tape stripping method. In rats, the gel containing 4% DEET resulted in a twofold increase in the permeation of MTX into the muscle over the 4 h period postdose. A modified HPLC method with a linear calibration curve (r > 0.999) over the range of 2-50 ng mL-1. quantitation limit of 0.5 ng mL-1, and mean recovery of approximately 87% was used for the quantitation of MTX in the tissue and fluid samples. PMID- 9210980 TI - p-Hydroxymethamphetamine enantiomer pharmacokinetics and metabolism in rats: absence of N-demethylation. AB - p-hydroxymethamphetamine (OHMAP) is one of the major metabolites of the widely abused drug methamphetamine (MAP). The demethylation of OHMAP to p hydroxyamphetamine (OHAP) has been shown in vitro but has never been reported in vivo. The disposition kinetics as well as the metabolism of OHMAP was investigated employing a sensitive HPLC method which can separate the enantiomers of OHMAP and OHAP. Both conjugated and unconjugated forms of these compounds can be quantitated. Male Sprague-Dawley rats were given an iv bolus of racemic OHMAP (20 mg kg-1) and serum and urine samples were collected at selected times. The serum concentration-time data for OHMAP enantiomers could be described by a biexponential equation. The clearance of D-OHMAP (93.5 mL min-1 kg-1) was slightly, but statistically significantly, greater than that of the L-enantiomer (83.9 mL min-1 kg-1). The steady-state volumes of distribution of L- and D-OHMAP were (mean +/- SD) 3.15 +/- 0.84 and 4.23 +/- 1.76 L kg-1, respectively. No significant concentrations or amounts of OHAP enantiomers could be detected in any serum or urine sample. Rats excreted more unchanged L-OHMAP (34%) than D OHMAP (29%). In contrast, more conjugated D-OHMAP (57%) was recovered compared to the conjugated L-OHMAP (52%). The results suggest that there is slight stereoselectivity in the disposition of OHMAP enantiomers. The N-demethylation product (OHAP) was not produced in vivo. PMID- 9210981 TI - The pharmacokinetics of 1954U89, 1,3-diamino-7-(1-ethylpropyl)-8-methyl-7H pyrrolo-(3,2-f)quinazoline, in dogs and rats after intravenous and oral administration. AB - 1954U89, 1,3-diamino-7-(1-ethylpropyl)-8-methyl-7H-pyrrolo-(3, 2-f)quinazoline, is a potent, lipid-soluble inhibitor of dihydrofolate reductase. The pharmacokinetics and bioavailability of 1954U89 were examined in male beagle dogs and male CD rats. Dogs received single intravenous (2.5 mg kg-1) and oral (5.0 mg kg-1) doses of 1954U89 with and without successive administration of calcium leucovorin. Single intravenous (5.0 mg kg-1) and oral (10 mg kg-1) doses of [1,3 14C2]1954U89 were administered to rats. Plasma concentrations of total radiocarbon were determined by scintillation counting, and intact 1954U89 was measured by HPLC. The mean plasma half-life was 3.2 +/- 0.62 and 4.2 +/- 0.68 h after intravenous and oral administration, respectively, to dogs. The pooled plasma half-life after intravenous administration to rats averaged 1.2 h; a reliable plasma half-life value after oral administration could not be determined. Mean total-body clearance was 2.4 +/- 0.39 and 4.5 +/- 1.1 L h-1 kg-1 after intravenous and oral administration, respectively, to dogs, and averaged 12 and 77 L h-1 kg-1 after intravenous and oral administration, respectively, to rats. Neither clearance nor bioavailability of 1954U89 in dogs was affected significantly by administration of calcium leucovorin. Absolute bioavailability was 54 +/- 12% in dogs and 16% in rats. PMID- 9210982 TI - off absorption, pharmacodynamics, metabolism and excretion of 14C-sumatriptan following intranasal administration to the beagle dog. AB - The pharmacodynamics, pharmacokinetics, metabolism, and excretion of 14C sumatriptan have been studied in the beagle dog following administration by the intranasal and other routes. The pharmacological response which was monitored, an increase in carotid arterial vascular resistance, correlated with the plasma levels of unchanged sumatriptan following intranasal, intravenous, or intraduodenal administration to the anaesthetised dog. The pharmacokinetics and metabolism of sumatriptan were then confirmed in conscious male and female dogs. Intranasal administration of 14C-sumatriptan resulted in rapid absorption of part of the dose. The overall bioavailability of sumatriptan was 40-50%. Sumatriptan was eliminated from plasma with a half-life of 1.5 or 1.9 h after intravenous or intranasal dosage respectively. Radioactivity was largely excreted in urine (up to 75% of the dose) with small amounts in the bile and faeces after intravenous and intranasal dosing, as sumatriptan and a major metabolite. The results from these studies suggest that intranasal administration provides a viable method for delivering sumatriptan to the systemic circulation. PMID- 9210983 TI - The influence of coffee with milk and tea with milk on the bioavailability of tetracycline. AB - The effect of milk added to coffee or black tea on the bioavailability of tetracycline was evaluated in 12 healthy volunteers according to a crossover design. Results showed that even a small volume of milk containing extremely small amounts of calcium severely impair the absorption of the drug, so that the presence of this metal ion should be carefully controlled in order to avoid decreasing the available tetracycline. PMID- 9210984 TI - Factor analysis of the Mayo-Portland Adaptability Inventory: structure and validity. AB - Principal-components (PC) factor analysis of the Mayo-Portland Adaptability Inventory (MPAI) was conducted using a sample of outpatients (n = 189) with acquired brain injury (ABI) to evaluate whether outcome after ABI is multifactorial or unifactorial in nature. An eight-factor model was derived which explained 64-4% of the total variance. The eight factors were interpreted as representing Activities of Daily Living, Social Initiation, Cognition, Impaired Self-awareness/Distress, Social Skills/ Support, Independence, Visuoperceptual, and Psychiatric, respectively. Validation of the Cognition factor was supported when factor scores were correlated with various neuropsychological measures. In addition, 117 patient self-rating total scores were used to evaluate the Impaired Self-awareness/Distress factor. An inverse relationship was observed, supporting this factor's ability to capture the two-dimensional phenomena of diminished self awareness or enhanced emotional distress. A new subscale structure is suggested, that may allow greater clinical utility in understanding how ABI manifests in patients, and may provide clinicians with a better structure for implementing treatment strategies to address specific areas of impairment and disability for specific patients. Additionally, more precise measurement of treatment outcomes may be afforded by this reorganization. PMID- 9210985 TI - Neuropsychological assessment after extremely severe head injury in a case of life or death. AB - A systematic neuropsychological assessment technique is described for use with severely physically disabled people who may be severely brain-damaged, in an incomplete locked-in state or potentially in vegetative state. The technique allows opinions regarding cognitive state to be statistically based. In the case described, the weight of expert opinion had been that involuntary feeding by gastrostomy tube should be terminated because the patient was functioning at a level little beyond the vegetative state, her quality of life was poor and she was unable to form a view about her present or future circumstances. An assessment approach is described which uses binomial statistics and allows for some variability in responding. Methods of minimizing sources of extraneous bias are also discussed. By use of this technique it was demonstrated that the patient was sentient though impaired, and that her own wish at the time of the assessment was to continue living. It is recommended that neuropsycholgical assessment of this kind should take place in all cases in which withdrawal of treatment is being considered and cognitive ability is not certain. PMID- 9210986 TI - Change in relationship status following traumatic brain injury. AB - Previous studies have highlighted the burden placed on family members and close partners of individuals who have sustained traumatic brain injury. This burden of stress has been attributed to the neurobehavioural sequelae of such injuries. However, the extent to which brain injury affects marriages and close relationships has never been statistically evaluated. This study looked at 131 adults with traumatic brain injury in order to determine the incidence of divorce/separation; 49 per cent of our sample reported that they had divorced or separated from their partners during a 5-8-year period following brain injury. Factors which may predict the outcome of relationships include severity of injury (as determined by length of post-traumatic amnesia), length of relationship, and time since injury. The influence of these factors in determining the risk of relationship breakdown is discussed. PMID- 9210987 TI - Use of a modified version of the Overt Aggression Scale in the measurement and assessment of aggressive behaviours following brain injury. AB - Aggressive behaviour creates a significant challenge in neurorehabilitation. Despite the success in using behaviour modification principles in the treatment of post-acute behavioural problems, psychopharmacological approaches to the management of aggression are more frequently reported. However, inconsistencies apparent in the literature hinder inter-study comparisons of treatment methods. These include severity of brain injury, neuropsychological status and rigour of experimental methodology used. Data about aggression is also inconsistently reported, especially with regard to classification and severity. Descriptions of how aggressive behaviour responded to pre-treatment is also generally absent. In this paper an observational rating scale is described in an attempt to address these inconsistencies. The Overt Aggression Scale has been modified by increasing the range of interventions to reflect current practice in neurorehabilitation, and by changing the language to make it suitable for UK users. A range of antecedents has also been added to make the scale useful in behavioural analysis. Preliminary results indicate inter-rater reliability is good, and it is a valid indicator of type and severity of aggression. Antecedents and interventions used in the management of aggressive behaviours in neurorehabilitation are also illustrated. Clinical use of the scale is also discussed. PMID- 9210988 TI - Awareness and perceptions of outcomes after traumatic brain injury. AB - Awareness or insight has been identified as a major factor in successful rehabilitation after traumatic brain injury (TBI). Anecdotal evidence suggests people with TBI are more likely to be aware of residual physical disabilities, perhaps focusing on these to the exclusion of other issues in the psychosocial and cognitive domains, for example. To investigate this more accurately, recovery and outcome questionnaires were administered to people with TBI and their nominated significant others, and, as appropriate, an assessment of their level of functioning was also recorded. Two- and three-way analyses (t-tests, Kendall's and Wilcoxon's) comparing these perceptions were then conducted. The results indicated a high level of agreement for basic demographic data and broad outcomes. It was found the subjects reported a lower rate of physical impairment and disability than the significant other or the author, suggesting that, as a group, they do not fixate on physical issues. Other areas of difference were found, such as a tendency for the significant other to perceive the subject as being more dependent in mobility and self-care tasks, possibly because of their close involvement. Also the author reported more impairments, using clinical language and assessment that did not necessarily have meaning or significance for the other groups. There was also evidence to support the notion that there is an inherent hierarchy of needs ranging from the lower-order, physiological or survival skills through to higher-order, self-actualizing areas. Because of the differing awareness and perceptions, care must be taken in service provision to identify the personal needs and values of each individual involved. PMID- 9210989 TI - Psychomotor agitation following gabapentin use in brain injury. AB - Gabapentin, an anticonvulsant structurally related to gamma-aminobutyric acid (GABA) was recently reported to be effective in pain associated with reflex sympathetic dystrophy (RSD) and in pain associated with neuropathy. Yet, to our knowledge, the use of gabapentin for neuropathic pain in the presence of cognitive impairment has not been reported. In this report, we describe two patients (one with a traumatic brain injury, one with a putative acquired brain injury) who presented to a neurorehabilitation unit complaining of pain that was diagnosed as neurologically mediated. Within one week of receiving a daily 900 mg dose of gabapentin, both patients complained of heightened anxiety and restlessness. Correspondingly, each reported a diminution of psychological symptoms within 48 hours of gabapentin cessation. These two cases suggest that gabapentin may cause agitation in cognitive impaired patients. Physicians treating brain-injured patients and prescribing gabapentin for neuropathic pain may wish to closely monitor patients for similar signs of restlessness or anxiety. PMID- 9210990 TI - The evolution of social attractiveness and its role in shame, humiliation, guilt and therapy. AB - This paper suggests that humans have innate needs to be seen as attractive to others. These needs form the basis for shame and mediate evaluations of social standing (status), social acceptance and social bonds. Shame and humiliation are associated with attacks on, and losses of, social attractiveness. The internal experiences of shame are derived from submissive strategies where one seeks to signal to others awareness of loss of social standing and limit possible damage. However, it is suggested that shame and humiliation differ from each other in a number of ways. For example, in shame the focus is on the self, while in humiliation the focus is on the harm done by others. Variations in the defensive strategies of shame and humiliation (e.g. avoidance, escape versus aggression and revenge) can pose particularly difficult problems in therapy. A focus on the role of social attractiveness in shame also allows for important distinctions to be drawn between shame and guilt. PMID- 9210991 TI - Interpersonal problems among patients suffering from panic disorder with agoraphobia before and after treatment. AB - The aim of this study was to examine interpersonal problems among panic disorder with agoraphobia patients before and after treatment. Patients (N = 46) suffering from panic disorder with moderate or severe agoraphobia and considering agoraphobia as their main problem were randomly assigned to receive either cognitive therapy or guided mastery therapy in a six-week in-patient group programme. The Inventory of Interpersonal Problems (IIP) and various symptom measures were administered at pretreatment and at one-year follow-up. Two IIP subscales were derived from factor analysis of the present data: affiliation problems and power problems. The overall pattern of results supported a state model of interpersonal problems. At pre-treatment, the scores on the affiliation problems subscale were clearly related to non-specific state characteristics, that is, to depression and general anxiety. From pre-treatment to follow-up, levels of interpersonal problems decreased significantly. Pre-treatment depression was a powerful predictor of change in interpersonal problems from pre treatment to one-year follow-up. On the other hand, interpersonal problems at pre treatment failed to predict the change in levels on various symptom scales. PMID- 9210992 TI - Natural cognitive coping strategies in schizophrenia. AB - Recent attention has focused on cognitive-behavioural treatment (CBT) for psychotic thinking. These interventions may be especially appropriate for drug resistant patients or as an adjunct to pharmacotherapy. However, CBT for schizophrenia was developed in the absence of any systematic investigation of personal self-statements that psychotic individuals develop on their own. The current investigation explored the natural cognitive strategies of 10 community based persons with schizophrenia. An exploratory interview was employed as the method of inquiry. A data bank of 344 statements was obtained from which 55 pertained to coping strategies. The results were interpreted using the grounded theory method (GTM) of qualitative analysis. Systematic analysis of the meaning units yielded a major category called coping self-talk which pertained to cognitive strategies (the focus of this research report). This category was composed of nine lower-level categories; that is, nine types of self-talk that persons with schizophrenia use actively in their own efforts toward managing psychotic symptoms were identified. These naturalistic coping strategies could provide useful guides for directing cognitive interventions. PMID- 9210993 TI - Signs and voices: joining a conversation in progress. AB - This commentary joins a conversation between Ryle and Leiman, which represents a larger conversation among theories of psychotherapy, semiotics and linguistics. The emerging collaborative understanding involves viewing people not as separate, unitary individuals but as mosaics or communities of different voices. The voices share their experiences by sign-mediated communication and engage in joint action. Projective identification may be considered as joint action mediated by non-verbal signs exchanged without awareness, in which the dyad becomes an effector for voices within both participants. The concepts of active voices and meaning-accumulating signs overcome the cognitive fallacy-the misleading notion that information in people is passive. PMID- 9210994 TI - Dementia: an intimate death. AB - This paper focuses on the loss of intimacy and mutuality in the relationship when one partner has Alzheimer's disease and suggests that the paradigm of anticipatory grief described in terminal care work may need to be reconsidered in relation to organic brain disease. An attempt is made to link some of the psychoanalytic literature of grief and the experience of clinicians in old age psychiatry working with patients and their relatives. Mention is made of implications for management. The paper includes illustrative clinical material. PMID- 9210995 TI - Features associated with speaking in tongues (glossolalia). AB - Reports of the frequency, context, associated behaviours, feelings and meaning associated with glossolalia were collected from three groups of informants: speakers (N = 14, who practised glossolalia), witnesses (N = 15, who had witnessed but had never practised glossolalia), and controls (N = 16, who had neither witnessed nor practised glossolalia). All informants were practising Christians. Speakers reported glossolalia as a regular, daily, private activity, usually accompanying mundane activities, as a special form of prayer associated with calm, pleasant emotions. By contrast, witnesses and controls were more likely to describe glossolalia as an exceptional activity, usually occurring in the religious group, and associated with excitement. The views of witnesses were closer to those of speakers than were the views of controls. It is suggested that there may be two types of glossolalia, of which one is more likely to be associated with psychopathology. PMID- 9210996 TI - Procedures as dialogical sequences: a revised version of the fundamental concept in cognitive analytic therapy. AB - Problematic action patterns that patients are unable to abandon or modify are important treatment targets in cognitive analytic therapy. They are called procedural sequences in the conceptual model underlying the approach. Interpersonal events in the patients' lives, and in the consulting room as well, frequently display such patterns. They are called reciprocal role procedures. In the present paper the sequential description of problem procedures will be examined by using Bakhtin's dialogical understanding of mental phenomena. A restatement of the original concept will be presented with the aim of integrating the sequential and reciprocal aspects of problematic action patterns. The revised concept of dialogical sequences will be illustrated by a number of case vignettes. The analysis of dialogical sequences provides a clear conceptual basis for early description of problem procedures in therapy. It also provides an effective tool for a detailed supervision of therapy sessions. Finally, dialogical sequence analysis may be used as a psychotherapy process research method to examine the interpersonal patterns in patient narratives. PMID- 9210997 TI - Losing ground: NIH funding to New York State researchers. PMID- 9210998 TI - Reinvigorating New York's academic biomedical research enterprise. PMID- 9210999 TI - Preparing physicians for careers in primary care internal medicine: 17 years of residency experience. AB - The objective of this survey was to demonstrate whether a primary care track internal medicine residency program emphasizing community-based health care of the urban sick poor trains physicians who will continue to practice in general internal medicine or similar fields. Thirty-five primary care residents (100% of graduates) who trained from 1976 through 1993 in the Adult Primary Care Track of the Internal Medicine Residency Program at St. Vincent's Hospital, New York were used as participants. PMID- 9211000 TI - The epidemiology and causes of injuries resulting in hospitalization in New York City: 1990-1992. AB - The purpose of this study is to present data on the distribution and etiology of nonfatal injuries resulting in hospital discharges in New York City (NYC). Records of all NYC residents discharged for injuries from acute stay hospitals 1990-1992 were tabulated. Injuries from surgical and medical procedures, adverse effects of drugs in therapeutic use, and late effects of injury were excluded. The results indicate that there was a marked geographic variation in rates: higher rates of assaults, self-inflicted injuries, burns, unintentional injuries from firearms, and injuries to bicyclists in disadvantaged neighborhoods. The data show that injuries in NYC have distinctive features that should form the basis for targeted prevention activities. PMID- 9211002 TI - Children's exposure to traffic and risk of pedestrian injury in an urban setting. AB - Pedestrian injuries to children represent a major urban health problem in the United States. Thousands of children each year are struck by moving motor vehicles; such collisions result in numerous hospitalizations and deaths. At particular risk are young school-age children between the ages of 5 and 9 years. Using a survey methodology, we collected data regarding the method by which children in an urban setting travel to and from school, in addition to the number of streets they cross in a typical school day. This information was compared with data from police records on street intersection locations of pedestrian collisions. There is a wide variation in the number of streets children cross in 1 day, calculated as the number of streets crossed in the entire day, not only those crossed to and from school. Children whose parents own a car and home cross an average of 3.7 streets per day, whereas children whose parents do not own both a car and home cross an average of 5.4 streets per day; this difference is highly significant (P < 0.0001). The largest differences in traffic exposure are between families reporting car- and-home ownership (x = 3.70 streets) versus those who do not own both a car and home (x = 5.39 streets) (Mann-Whitney = -5.5, P < 0.0001). There is a significant correlation between the proportion of children driven home from school and the rate of pedestrian injury in different regions of Baltimore. In areas where children are driven home, rates of pedestrian injury are significantly lower, whereas in areas where children walk home, rates of pedestrian injury are high (r = -0.79, P < 0.01). This study underscores the importance of adapting the child's environment to prevent injury. Interventions that alter the nature of the hazard are indicated. Changing the environment may ultimately prove more useful than attempting to change children's behavior. PMID- 9211001 TI - Prenatal care utilization in New York City: comparison of measures and assessment of their significance for urban health. AB - This paper considers policy and programmatic consequences of shifting measurement of prenatal care utilization from the Kessner Index (KI) to the Adequacy of Prenatal Care Utilization Index (APNCUI). In gauging the adequacy of prenatal care utilization, the KI considers the timing of prenatal care initiation and the number of prenatal visits. The APNCUI also considers both timing of initiation and number of visits, but the approach taken to conceptualizing and measuring these two aspects of prenatal care utilization is more refined. We used birth certificates to calculate the KI and the APNGUI for 217,183 New York City (NYC) births in 1991-1992. We used cross-tabulations and bivariate odds ratios to compare the classifications resulting from the respective indexes. The APNCUI detected some important dimensions of the problem of inadequate prenatal care use that are not evident when using the KI. The proportion of births with inadequate use increases from 18% with the KI to 35% with the APNGUI. Groups of women at elevated risk for inadequate use are the same, but the KI understates significantly the risk for Hispanic women, teens, women who are less well educated, and those on WIC and Medicaid. The APNGUI yields a fuller picture of the degree to which some urban women are at risk for inadequate prenatal care use. Use of the APNGUI in quality assurance, monitoring, and research is recommended. PMID- 9211003 TI - The working child and the street child: effect on future child development. PMID- 9211004 TI - Methods for successful follow-up of elusive urban populations: an ethnographic approach with homeless men. AB - Public health is paying increasing attention to elusive urban populations such as the homeless, street drug users, and illegal immigrants. Yet, valid data on the health of these populations remain scarce; longitudinal research, in particular, has been hampered by poor follow-up rates. This paper reports on the follow-up methods used in two randomized clinical trials among one such population, namely, homeless men with mental illness. Each of the two trials achieved virtually complete follow-up over 18 months. The authors describe the ethnographic approach to follow-up used in these trials and elaborate its application to four components of the follow-up: training interviewers, tracking participants, administering the research office, and conducting assessments. The ethnographic follow-up method is adaptable to other studies and other settings, and may provide a replicable model for achieving high follow-up rates in urban epidemiologic studies. PMID- 9211005 TI - The position of the New York Academy of Medicine on physician-assisted suicide. AB - In January 1997, after a lengthy, careful, and difficult process, an ad hoc group, chaired by Dr. Alan R. Fleischman, a Senior Vice President of the New York Academy of Medicine (NYAM), with representation from clinical medicine, biomedical ethics, law, and the clergy, developed a position on the difficult and contentious issue of physician-assisted suicide. After substantial debate, the Board of Trustees of NYAM authorized a letter from the President of the Academy to the Justices of the United States Supreme Court and to the attorneys on both sides of the cases about to be argued before the Court. The text of that letter, which summarizes the views of the New York Academy of Medicine, is reproduced here. PMID- 9211006 TI - A physician's position on physician-assisted suicide. AB - On April 29, 1996, Dr. Quill offered testimony at an Oversight Hearing on "Assisted Suicide in the United States," before the Subcommittee on the Constitution of the House of Representatives Committee on the Judiciary. That testimony is reproduced here, with permission of the author. PMID- 9211007 TI - The epidemiology and prevention of rheumatic fever. 1952. PMID- 9211008 TI - The origins of the College of Physicians and Surgeons of Columbia University. PMID- 9211009 TI - Historical note and notice: The New York Society of Forensic Sciences at Lehman College, City University of New York, 1985 to 1996. PMID- 9211010 TI - Declining AIDS mortality in New York City. New York City Department of Health. PMID- 9211011 TI - Cardiovascular pharmacotherapy--from prevention to treatment. Proceedings of a symposium. Santiago, Chile. PMID- 9211012 TI - Hazards, risks, and threats of heart disease from the early stages to symptomatic coronary heart disease and cardiac failure. AB - The epidemiologic approach to investigation of atherosclerotic cardiovascular disease has provided many insights into the preclinical and clinical spectrum of the disease. The hazard of developing atherosclerotic cardiovascular disease is substantial with coronary heart disease (CHD), the most common and most lethal feature. The outlook in those who manage to survive the initial episode is also serious, with a 10-year mortality rate of 37% for persons with angina and a 55% rate for those sustaining a myocardial infarction. Fifteen percent of persons developing CHD present with a fatal event, and 38% of infarctions go unrecognized. The presence of atherosclerosis in one vascular territory imposes an increased risk of its appearing in another area at two to six times the general population rate. The major cardiovascular risk factors adversely affect all arterial vascular territories so that correction of risk factors targeted at one particular atherosclerotic outcome may also favorably influence the other risk factors. Coronary disease is the most prevalent lethal hazard of hypertension, dyslipidemia, glucose intolerance, and cigarette smoking. These risk factors cluster and optimal therapy must improve the whole risk profile. Women share the same risk factors for CHD as men. Although women have a lower absolute risk for most risk factors, a high total/HDL cholesterol ratio, left ventricular hypertrophy, and diabetes each tend to eliminate the female advantage. Menopause also promptly escalates risk threefold. Although women tend to have a lower incidence than men, the initial attack is just as highly lethal in women, and their subsequent outlook as survivors is at least as serious as for men. Sudden death is a pre-eminent feature of coronary disease and cardiac failure. Coronary disease increases sudden death risk 3.3-fold and cardiac failure 4.8-fold. Sudden death incidence varies in relation to the same cardiovascular risk factors as coronary heart disease, with no unique risk factors identified. However, multivariate combinations of these in a profile can identify high-risk candidates for sudden death as well as coronary attacks in general. The key to prevention of sudden death is to prevent coronary attacks and cardiac failure. Despite aggressive cardiac revascularization and treatment of hypertension, congestive heart failure (CHF) has not decreased in prevalence, and innovations in the treatments of overt failure have not substantially improved survival. Median survival is only 1.7 years for men and 3.2 years for women. The conditional probability of developing CHF can be estimated using a logistic function comprised of age, systolic pressure, vital capacity, heart rate, ECG left ventricular hypertrophy (LVH), glucose intolerance, x-ray enlargement, and presence of CHD and heart murmurs. Eighty percent of CHF events occur in persons in the upper quintile of multivariate risk. Continued clinical, metabolic, and epidemiologic research have expanded and refined atherosclerosis risk factors. The lipid connection is now concerned with the apoprotein makeup of the lipids, subfractions of lipids, and Lp(a). The diabetic influence is now focused on insulin resistance. Ambulatory monitoring is being used to evaluate blood pressure and silent ischemia. Fibrinogen and leukocyte counts have emerged as possible indicators of unstable lesions. Prospects for primary and secondary prevention are good if public health measures, health education, and preventive medicine are implemented based on existing knowledge of correctable or avoidable risk factors. The potential for more effective prevention continues to expand, and great advances have already been made in countries where aggressive preventive measures have been implemented to correct the major established risk factors. PMID- 9211013 TI - Oral nitrates: more than symptomatic therapy in coronary artery disease? AB - The predominant venodilator properties of the nitrates and their augmentation of collateral coronary blood flow to the ischemic myocardium endows them with some ideal characteristics for treating myocardial ischemic syndromes. Additional efficacy stems from the ability of the nitrates to replenish the deficient endothelium-derived relaxing factor (EDRF), nitric oxide (NO), in patients with coronary heart disease and also to inhibit platelet aggregation. In stable angina pectoris, the antianginal and antiischemic effects of oral nitrates are well established. Continuous administration of nitrates may lead to tolerance of their clinical efficacy. Recent studies, however, have demonstrated that when used in recommended doses, tolerance can be avoided during long-term treatment with oral nitrates without provocation of anginal attacks during periods of low nitrate levels at night and early hours of the morning. Thus, prolonged treatment with an asymmetric twice-daily regimen of immediate-release isosorbide-5-mononitrate in patients with stable angina pectoris does not give rise to clinical tolerance, prolongs exercise duration, and delays the onset of myocardial ischemia. In unstable angina pectoris, nitrates rapidly relieve chest pain and ameliorate the electrocardiographic signs of myocardial ischemia. In patients with acute myocardial infarction, early treatment with nitrates prevents left ventricular dilatation, improves pumping function, and reduces the risk of ventricular arrhythmias. In patients with chronic heart failure, oral nitrates improve exercise tolerance and, when given in combination with the systemic arterial dilator hydralazine, extend survival. Meta-analysis of published studies has demonstrated that both intravenous and oral nitrates reduced infarct size and morbidity and mortality in patients with acute myocardial infarction. In the ISIS 4 post-infarction study, isosorbide-5-mononitrate 60 mg once daily was not superior to placebo in reducing mortality risk. However, in the GISSI 3 study, the combination of nitrates with an angiotensin-converting enzyme (ACE) inhibitor reduced mortality risks by 17% in patients with acute myocardial infarction. In both the ISIS 4 and GISSI 3 studies, 62% and 57% of the patients in the placebo and control groups, respectively, were treated with nitrates for control of rest angina, myocardial ischemia, and or left ventricular failure symptoms, and this widespread use of open-label nitrates in the control groups may have diluted the true beneficial effects of nitrates in both studies. Taken together, these many studies with oral nitrate treatment in coronary heart disease and heart failure clearly emphasize that these drugs are safe and play more than a symptomatic role in the management of patients with acute and chronic ischemic syndromes due to coronary artery disease. PMID- 9211014 TI - Beta-blocking drugs and coronary heart disease. AB - Since the development of beta-adrenergic blocking agents over 30 years ago, they have been established as an important therapeutic modality in the treatment of coronary heart disease. This article reviews the role of beta-blockers in the treatment of hypertension, angina, acute myocardial infarction, and heart failure. A number of multicenter studies indicate that beta-blockers have an important effect in decreasing morbidity and mortality in patients with hypertension and appear to have a relatively increased importance in elderly patients with hypertension. Although their long-term effects on the mortality of angina pectoris have not been fully investigated, investigations indicate that, compared with other drugs, beta-adrenergic blocking agents significantly decrease the frequency and duration of angina pectoris. The largest use of these drugs has been examined in the treatment of acute myocardial infarction. Two major trials, the Norwegian Timolol Trial and the Beta Blocker Heart Attack Trial, confirm the long-term benefit in patients following acute myocardial infarction. Their use in heart failure is under close investigation following a series of preliminary studies suggesting they may decrease mortality risk. Although the tolerability of these drugs has been questioned, careful examination of clinical trials indicate that they are relatively well tolerated. These observations emphasize the importance of beta-adrenergic blocking agents in hypertension, angina, and acute myocardial infarction and speak to a wider clinical use of these drugs. PMID- 9211015 TI - Detection of myocardial damage by serial measurements of cardiac troponin T, CK MBmass, and TROPT rapid test. AB - Detection of cardiac damage is greatly facilitated by serial blood measurements of myocardial cell markers. In many hospitals creatine kinase MBmass concentration (CK MBmass) constitutes the biochemical criterion (WHO) for acute myocardial infarction (AMI). Cardiac troponin T (TnT) is an even more sensitive and specific marker for myocardial damage. With discriminator levels of 10.0 and 0.10 micrograms/l, respectively, serial measurements of both markers provide a useful diagnostic strategy for ischemic heart disease. This survey reviews representative cumulated time curves in individual patients covering the spectrum of myocardial damage, including unstable angina pectoris (UAP), non-Q-wave and Q wave infarctions with and without early reperfusion, re-infarction, and subacute infarction. Increased TnT detects minor myocardial damage (MMD) in over 30% of patients with UAP, although CK MBmass remains below its discriminator. Subacute infarction is detected by the wide diagnostic time window of the serum TnT at a time when CK MBmass has already returned to normal. In a substudy of 502 suspected cases of AMI, the distributions of maximum serum TnT concentrations within each patient series demonstrated that TnT had a diagnostic sensitivity of 100% and a specificity of 99%. Median, 5th and 95th percentiles of maximum TnT values within the diagnostic subgroups showed that serum TnT was increased five fold more than CK MBmass. Median values of Q-wave AMI were higher than in non-Q wave AMI. A diagnostic strategy using TROPT, a rapid test specific for the cardiac isoform of TnT with a detection limit 0.10 microgram/l, is presented. PMID- 9211016 TI - Thrombolytic therapy in acute myocardial infarction. AB - Thrombolytic trials in acute myocardial infarction have progressively increased in size, with the GUSTO study including over 40,000 patients. New thrombolytic drugs are unlikely to lead to further major reductions in fatality, and demonstrating small differences from existing compounds would require even larger trials to establish mortality benefits. New agents may, however, have advantages, such as a greater ease of administration or a reduced cost, and the problem is how to assess such new agents in the most efficient way. The concept of equivalence as a clinical trial endpoint is discussed, and the INJECT study, comparing reteplase and streptokinase in acute myocardial infarction, is described as a prototype equivalence trial. PMID- 9211018 TI - Retardation of coronary atherosclerosis: the Bezafibrate Coronary Atherosclerosis Intervention Trial (BECAIT) and other angiographic trials. AB - A large number of both primary and secondary preventive trials suggest that treatment of elevated plasma lipids may reduce the frequency of coronary heart disease (CHD) events. Meta-analyses indicate that for every 10% reduction of cholesterol, CHD mortality is lowered by 13% and all-cause mortality by 10%. Experience from several angiographic trials also suggests that coronary atherosclerosis can be retarded, and in some instances limited regression induced, by low-density lipoprotein (LDL)-cholesterol reduction and/or high density lipoprotein (HDL)-cholesterol elevation. Coronary angiographic studies have shown that although the effects on retardation/regression of altherogenic lesions have been small, with luminal diameter changes of around 0.10 mm, the effects on clinical events were more substantial, with reductions of the order of 50%. There is also evidence that it is the mild and moderate lesions that are of particular concern with respect to the occurrence of clinical coronary events. The progression of atherosclerosis and the occurrence of coronary events are probably not exclusively dependent on a lowering of LDL-cholesterol concentrations. Analyses from the Monitored Atherosclerosis Regression Study (MARS), the Cholesterol Lowering Atherosclerotic Study (CLAS), and the Programs on the Surgical Control of the Hyperlipidaemias (POSCH) indicate that triglyceride-rich lipoproteins may also be of importance for the progression of mild/moderate lesions in subjects treated with cholesterol-lowering regimens. Recently, the results of the 5-year Bezafibrate Coronary Atherosclerosis Intervention Trial (BECAIT) demonstrated that bezafibrate significantly retarded the progression of coronary atheroma and coronary events in young male survivors of myocardial infarction, as assessed by changes in minimum lumen diameter. This positive outcome is most likely due to the beneficial treatment effects on the levels of serum triglycerides (-31%), plasma fibrinogen (-12%), and HDL cholesterol (+9%). The results of the BECAIT on progression of coronary atherosclerosis are comparable with those of two recent angiographic trials with HMG-CoA reductase inhibitors, the Multicenter Anti-Atheroma Study (MAAS), and the Regression Growth Evaluation Statin Study (REGRESS), despite different lipid and metabolic effects. BECAIT is the first controlled angiographic study to show that a fibrate can significantly retard the progression of coronary atherosclerosis. The exact mechanisms by which this occurs remain to be elucidated. However, the results of BECAIT illustrate the importance of factors other than LDL cholesterol in the progression of coronary atherosclerosis. PMID- 9211017 TI - Pharmacology of carvedilol: rationale for use in hypertension, coronary artery disease, and congestive heart failure. AB - Carvedilol is a novel, multiple-action cardiovascular drug that is currently approved in many countries for the treatment of hypertension. The reduction in blood pressure produced by carvedilol results primarily from beta-adrenoceptor blockade and vasodilation, the latter resulting from alpha 1-adrenoceptor blockade. These actions, as well as several of the other activities of carvedilol, are associated with cardioprotection in animal models that occurs to a degree that is greater than that observed with other drugs. The multiple actions of carvedilol may also provide the underlying rationale for the use of the drug in the treatment of coronary artery disease and congestive heart failure. By virtue of being both a beta-blocker and a vasodilator, carvedilol significantly decreases myocardial work by reducing all three components of myocardial oxygen demand, namely, heart rate, contractility, and wall tension. The vasodilatory effects of carvedilol reduce afterload, and the resulting decrease in impedance to left ventricular ejection offsets the negative inotropic effect that would normally result from beta-blockade. As a consequence, stroke volume and cardiac output are maintained or even increased in animals and in patients with congestive heart failure who are treated with carvedilol. Carvedilol and several of its metabolites are potent antioxidants, and this activity may account, in part, for the cardioprotective effects of the drug observed in animal models of acute myocardial ischemia and, in theory, could also serve to protect the myocardium of patients with hypertension, coronary artery disease, and congestive heart failure, in which oxidative stress is now recognized to occur. The antioxidant effects of carvedilol may both inhibit the direct cytotoxic actions of reactive oxygen radicals and prevent oxygen-radical induced activation of transcription factors and genes associated with inflammatory and remodeling processes. Accordingly, carvedilol inhibits the gene expression of the intracellular adhesion molecule-1 (ICAM-1), an adhesion molecule for polymorphonuclear leukocytes, which typically infiltrate the myocardium under conditions of ischemia and may exacerbate ischemic injury. The antioxidant activity of carvedilol has been shown to inhibit the oxidation of low density lipoprotein (LDL) in vitro, thereby preventing the formation of this cytotoxic and atherogenic form of LDL. It follows, therefore, that in animal models of hyperlipidemia, carvedilol attenuates aortic lipid accumulation and decreases the aortic content of monocytes and foam cells, and at the same time it has been shown to preserve endothelial integrity and function. These actions of carvedilol are not shared by other beta-blockers or by other drugs currently used in the management of hypertension, coronary artery disease, or congestive heart failure. The multiple actions of carvedilol may provide the underlying pharmacologic rationale for the use of this drug in the treatment of patients with coronary artery disease or congestive heart failure, and these actions may account, at least in part, for the reduction in mortality produced by carvedilol in clinical trials involving patients with congestive heart failure. Likewise, these actions of carvedilol may also provide protection, beyond that afforded from reduction in blood pressure, against secondary organ damage in hypertensive patients treated with the drug. PMID- 9211019 TI - Origins of symptoms in heart failure. PMID- 9211020 TI - Diuretics in the prevention and treatment of congestive heart failure. AB - Diuretics, either as monotherapy or in combination with other drugs in the management of hypertension, have retarded the development of left ventricular hypertrophy and congestive heart failure in many patients. Diuretics also remain one of the most effective therapies in the treatment of congestive heart failure, when given with an ACE inhibitor and digoxin. PMID- 9211021 TI - Digoxin therapy in chronic heart failure. AB - Digitalis has been used for more than 250 years, but its role in the treatment of chronic heart failure has been intensively investigated only during the past two decades. Digoxin increases cardiac output both at rest and during exercise, alone or in combination with ACE inhibitors, and these hemodynamic effects are sustained during chronic therapy. A daily dose of digoxin that achieves a serum concentration of approximately 1.2 ng/ml is associated with a significant improvement in central hemodynamics, particularly in patients with impaired cardiac function despite pretreatment with diuretics and ACE inhibitors. Acute administration of digoxin in patients with chronic heart failure has an immediate sympathoinhibitory effect, and chronic therapy is associated with a sustained decrease in serum norepinephrine concentration. Discontinuation of digoxin in patients with chronic heart failure resulted in hemodynamic deterioration, which was reversed when the drug was readministered. Randomized withdrawal of digoxin in patients receiving only diuretics (PROVED study), or its withdrawal in patients receiving diuretics and ACE inhibitors (RADIANCE study), was associated with worsening of the clinical evidence of heart failure and a decrease in left ventricular systolic function in both studies. In the only large-scale, placebo controlled mortality study reported thus for (DIG Trial), 7788 patients received standard drug treatment for chronic heart failure in addition to either digoxin or placebo. Digoxin had no impact on survival over the 37 months of follow-up, but the incidence of hospitalizations due to worsening heart failure was significantly reduced in patients receiving the drug compared with those receiving placebo. PMID- 9211022 TI - ACE inhibitors in heart failure: prospects and limitations. AB - ACE inhibitors have been shown to be effective in reducing the morbidity and mortality of patients with left ventricular systolic dysfunction, but their application to clinical practice in this situation is still limited. In part, the failure to prescribe an ACE inhibitor to a patient with left ventricular systolic dysfunction is due to perceptions regarding their side effects, such as cough and renal dysfunction. Relatively few patients with left ventricular systolic dysfunction and a serum creatinine > or = 2 mg/dl receive an ACE inhibitor in clinical practice. In this situation one should consider an agent such as fosinopril, which is metabolized by the liver as well as secreted by the kidney. In patients with moderate renal dysfunction, fosinopril has been well tolerated without an increase in serum creatinine. In patients who develop cough due to an ACE inhibitor, consideration should be given to an angiotensin II type 1 receptor blocking agent, such as losartan. The relative safety and efficacy of an ACE inhibitor compared with an angiotensin II type 1 receptor blocking agent is being explored in a prospective randomized trial (Evaluation of Losartan In The Elderly [ELITE]), as well as the safety and pharmacological effectiveness of adding an angiotensin II receptor antagonist to an ACE inhibitor (Randomized Angiotensin receptor antagonists-ACE-inhibitor Study [RAAS]). There may also be a role for the combination of an aldosterone receptor antagonists and an ACE inhibitor in patients with left ventricular systolic dysfunction. Once an ACE inhibitor is administered to a patient with left ventricular systolic dysfunction it should be continued indefinitely. ACE inhibitors may be of value not only in preventing the progression of heart failure but also in reversing endothelial dysfunction and preventing the development of atherosclerosis and its consequences, such as myocardial infarction. PMID- 9211023 TI - Second- and third-generation beta-blocking drugs in chronic heart failure. AB - The left-ventricular (LV) functional, hemodynamic, and antiadrenergic effects of metoprolol, bucindolol, and carvedilol have been compared in three concurrent placebo-controlled clinical trials in patients with symptomatic idiopathic dilated cardiomyopathy. All three drugs were well tolerated, all produced at least moderate degrees of beta-blockade as assessed by reduction in exercise heart rate, and all increased the left-ventricular ejection fraction. Compared with the beta 1-selective, second-generation compound metoprolol, the third generation compounds bucindolol and carvedilol lowered indices of adrenergic activity and tended to improve LV function to a greater extent. In patients with chronic heart failure there may be important therapeutic response differences between second- and third-generation beta-blocking agents. PMID- 9211025 TI - Digital reconstruction with island flaps. AB - Better understanding of the vascular anatomy of the hand and of flap perfusion allows the hand surgeon to perform single-stage reconstruction of digital defects through a multitude of island flap transfers. The usefulness of more than 20 separate island flaps is discussed, and the technique of flap transfer is presented for each. PMID- 9211024 TI - Combination drug therapy in chronic heart failure: is treatment part of the problem in heart failure? AB - Despite advances in medical treatment, the annual mortality associated with severe heart failure remains over 40%, and even in mild heart failure the associated mortality is 40% over 4 years. Once it has been demonstrated that the morbidity and mortality to heart failure can be adequately addressed by combinations of drug therapy, then it is logical to attempt to strip out redundant components of these therapeutic regimes. In the meantime, however, combination therapy is required to counter many of the pathophysiological facets of the heart failure syndrome, including fluid retention, neuroendocrine activation, progressive ventricular dysfunction, and sudden cardiac death. Diuretics and ACE inhibitors are well-established drug treatments. Digoxin appears to lessen the rate of progression of heart failure without altering survival. New evidence suggests that beta-blockers may be useful additions to the heart failure therapeutic armamentarium, although whether all beta-blockers are equally effective remains to be established. PMID- 9211027 TI - Microvascular reconstruction of the distal digits by partial toe transfer. AB - Advances in microsurgery have provided multiple options for reconstruction of distal digital deficit using various parts of the finger. These relatively minor microsurgical procedures always lead to a great patient satisfaction both in function and appearance. PMID- 9211026 TI - Reconstruction of the hand with forearm island flaps. AB - From all of the flaps reviewed, it is important to know how to select the most suitable choice in each case. Aside from the technical expertise of the surgeon, the indication depends on the size and the location of the substance loss. For large defects in any location, the radial forearm flap remains the most reliable and safest choice. For children and women, the authors prefer distant pedicled transfers or free flaps to minimize cosmetic donor site morbidity. For small or medium defects that cannot be managed by a local transposition flap, the indication is based on the location of the wound. Palmar defects, if proximal and ulnar, may be covered using the dorsal ulnar flap, with little morbidity in the donor area. The anterior interosseous flap seems a better choice whenever vascularized tendon, nerve, or bone are needed also. For the first web space and neighboring radial defects, the posterior interosseous flap provides a reasonable alternative. Dorsal defects of the hand can be reconstructed with a posterior interosseous flap, provided there is no suspicion of injury to the anastomotic dorsal system of the wrist. The anterior interosseous flap is a good choice for composite osteocutaneous transfers. For complex composite defects, the ulnar artery forearm flap distally based may be indicated for reconstructive problems requiring vascularized flexor tendons. The anterior interosseous flap is able to provide excellent quality vascularized bone. Indications depend above all on the surgeon's experience and on the different schools. As always, the better flap is that which is performed by the surgeon who has mastered the particular surgical technique. In conclusion, this article is devoted to an update on forearm flaps and illustrates the innovative strength of this specialty. It also points out that, through in depth knowledge of the anatomy, flaps may be raised from many anatomic regions of a limb without disturbing the main vascular axis of that extremity. Minimizing the donor site morbidity while maximizing the quality of the reconstruction is the primary concern when indications are established for reconstructive hand surgery, which is where one of the authors' main research efforts resides. PMID- 9211028 TI - Free flap coverage of the hand. AB - Microvascular free tissue transfer has been a major advancement in the treatment of soft-tissue defects of the hand. Free tissue transfers have expanded our options and have altered our approach to hand defects. It is no longer satisfactory to cover hand wounds with unsightly, bulky flaps of tissue. Microsurgical free tissue transfers have given us the tools for more refinement in hand soft-tissue reconstruction and have changed the standards for a successful outcome. PMID- 9211029 TI - Advances in peripheral nerve repair. AB - The basic tenets of peripheral nerve repair as outlined in the introduction remain valid. Advances in the field of peripheral nerve repair have become focused on enhancement of the rate, completeness, and accuracy of neural regeneration on a cellular and molecular level. It is hoped that, within the next decade, techniques that have shown promise experimentally will become mainstays of treatment. PMID- 9211031 TI - Advances in digital replantation. AB - Technical advances in revascularization and in skin cover have resulted in increases in the possibilities of digital replantation. In cases of multiple digital lesions and isolated avulsions of the thumb, all attempts to ensure recovery of basic pinch capacity are justified. On the other hand, in cases of proximal amputations with extensive avulsion or crush injury such attempts should be abandoned if results on function are important. Primary and secondary surgery must confront two difficult problems namely, repair of the avulsed flexor tendons and resensibilization. But in the field of replantation one should not be too dogmatic because for cosmetic reasons, some patients may find even a stiff or insensitive finger satisfactory. Distal and very distal replantations yield the best results in all fields. This information should be brought to the attention of all trauma teams that tend to consider such replantations useless. These replantations are both rapid and simple, requiring only mastery of suturing vessels 0.5 mm in diameter. The surgeon responsible for replantation should weigh the indications after assessing the patient's needs and explaining the advantages and drawbacks of this type of surgery and its results on function. PMID- 9211030 TI - New reconstructive procedure for brachial plexus injury. AB - The double free-muscle transfer technique has restored prehension in patients following complete avulsion of the brachial plexus. This achievement was almost inconceivable as recently as several years ago and has now given new hope for these patients to be able to use their otherwise useless limbs. PMID- 9211032 TI - Management of severe ischemia of the upper extremity. AB - Ischemia of the upper extremity is unusual, particularly in patients without trauma or iatrogenic injury to the vessels of the arm. Some authors have suggested that ischemia of the upper extremity is one sixth as common as that in the leg. Management of patients with vascular trauma or iatrogenic injury is usually straightforward, with direct repair or vein grafting of the injured vessel. Patients with ischemia from vascular disease, however, present a different set of management problems. These individuals often suffer from systemic medical problems that lead to their vascular disease, which is often very severe and accelerated because of the underlying cause. This article discusses the approach to these patients and options for management. PMID- 9211033 TI - Management of vasospastic disorders of the hand. AB - Vasospastic disorders of the upper extremity are common and often difficult to treat. Using the proposed classification system (Table 2) allows management based upon pathologic condition, physiologic staging, and response to treatment. Identifying patients in this way also helps in determining which treatments are most appropriate. The basic approach to management includes environmental and behavioral modifications including cessation of tobacco use, protection of hands, and avoidance of situations that trigger the vasospastic response. Pharmacologic therapy may provide good results in a majority of patients. Surgical intervention is reserved for patients with vaso-occlusion, ischemia, and refractory symptoms in spite of attempts at medical management. Surgical options include vascular reconstruction, peripheral sympathectomy, or a combination of techniques. The goal of medical and surgical management is to increase total or nutritional blood flow in the digits. PMID- 9211034 TI - Comprehensive management of Raynaud's syndrome. AB - Raynaud's syndrome stands as a landmark to our ignorance of the mechanisms involved in the interrelationship of the central nervous system and peripheral circulatory physiology. Proper management for these unfortunate patients demands an integrated comprehensive effort of the pharmacologic, physical medical, and surgical disciplines. PMID- 9211035 TI - Update on tendon repair. AB - The past decade has seen some remarkable advances in the management of acute tendon repair, both flexors and extensors. New surgical techniques, such as the epitenon-first technique for flexor tendons, combined with early motion rehabilitation, pharmacologic intervention to prevent adhesions, and noninvasive imaging techniques such as MR imaging to assess repair integrity lead to one inescapable conclusion. Continued evolution in attempts to improve the overall results of tendon injury can only mean that hand surgeons have not yet reached the perfect solution in tendon repair strategy. Much progress has been made since the era of "no man's land" when primary repair in zone II was in disfavor, but many challenges remain to be met if the "perfect" tendon repair is to be realized. PMID- 9211036 TI - Advances in reconstruction of digital joints. AB - The recent development of dynamic traction provides several advantages for the treatment of intra-articular fractures of the hand: Ligamentotaxis reduces fracture fragments and realigns joint surfaces, Contracture of joint ligament and periarticular structures is prevented, Collapse of fracture fragments is prevented, Cartilage healing and regeneration are enhanced, Joint mobility is retained, Extensive surgery may be avoided, As Leonardo da Vinci stated, "To understand motion is to understand nature." PMID- 9211037 TI - Outcomes assessment in hand surgery. What's new? AB - Outcomes of medical care include many dimensions: physical, social, and emotional function; symptoms; and satisfaction. In many cases, these dimensions can be assessed by patient-completed questionnaires with high reliability, sensitivity, and responsiveness. One such questionnaire-the upper extremity disabilities of arm, shoulder, and hand (DASH)-is reviewed. PMID- 9211038 TI - Organizational development strategies for integrating mental health services. AB - The recent debates about health care reform have focused attention on the need to develop organized systems of care capable of delivering comprehensive services which are coordinated or integrated. Achieving service integration has emerged as a central and pressing objective in most mental health systems in response to existing difficulties with fragmentation of care. However, attempts at service integration often fail at the implementation stage as provider agencies zealously guard their organizational boundaries and struggle with each other for power and control. In this article, the authors formulate an organizational development approach to service integration that focuses on reducing the rigid maintenance of agency boundaries by developing informal networks among staff of local provider agencies. Eight strategies, drawn from the research literature on services integration and recently implemented by a local mental health authority, are described as potential tools for use by systems managers in accomplishing these goals. PMID- 9211039 TI - A four-year comparison of physical health benefit expenditures of mental and physical health claimants. AB - The pattern of covered physical health benefit expenditures before and after initiation of mental health treatment for a cohort of individuals with mild to moderate mental health problems was examined in this study. The results indicated the mental health treatment group (MHTG) began to have higher total covered benefits expenditures about one year prior to starting mental health treatment. Statistically significant differences between the MHTG and comparison groups (groups with no mental health treatments during the study period and matched on age and gender) began at the quarter just before treatment and continued through the seventh quarter after treatment initiation. The higher pattern of total health benefit expenditures of the MHTG declined gradually for two years before it returned to pretreatment levels. The study suggests the need for a better understanding of the types of physical health benefits expenditures preceding and following mental health treatment. It further indicates that the integration of physical and mental health benefits case management may have potential for reducing overall health benefit costs. PMID- 9211040 TI - Bridging the gap between service need and service utilization: a school-based mental health program. AB - In an effort to bridge the gap between service need and service utilization, an urban based, university affiliated children's psychiatric outpatient clinic has implemented a program which provides mental health services in inner city schools. When compared with the central clinic populations (N = 304), the school sample (N = 44) was markedly socioeconomically disadvantaged, minority, and as psychiatrically impaired as the central clinic population. School based mental health services have the potential for bridging the gap between need and utilization by reaching disadvantaged children who would otherwise not have access to these services. Implications for such services are discussed. PMID- 9211041 TI - Is the Addiction Severity Index a reliable and valid assessment instrument among clients with severe and persistent mental illness and substance abuse disorders? AB - OBJECTIVE: This study examined aspects of reliability, validity and utility of Addiction Severity Index (ASI) data as administered to clients with severe and persistent mental illness (SMI) and concurrent substance abuse disorders enrolled in a publicly-funded community mental health center. METHODS: A total of 62 clients with SMI volunteered to participate in an interobserver and test-retest reliability study of the ASI. Spearman-Brown and Pearson correlation coefficients were calculated to examine the extent of agreement among client responses. RESULTS: Overall 16% of the composite scores could not be calculated due to missing data and 31% of the clients misunderstood or confused items in at least one of the seven ASI domains. As a whole, the interobserver reliability of the ASI composite scores for those subjects where sufficient data were available was satisfactory. However, there was more variance in the stability of client responses, with four composite scores producing test-retest reliability coefficients below .65. CONCLUSION: Evidence from this study suggests that the ASI has a number of limitations in assessing the problems of clients with severe and persistent mental illness, and it is likely that other similar instruments based on the self-reports of persons with severe and persistent mental illness would also encounter these limitations. PMID- 9211042 TI - School violence reduction: a model Jamaican secondary school program. AB - Violence in United States' schools is epidemic. Solutions are rare. Community mental health centers are now being challenged to become part of the solution. The Montego Bay Secondary School project presents an example of how violence reduction can be achieved using almost no physical resources and the special effect, called the "Bruno Effect", created by one Jamaican police officer with the consultation of a psychodynamically-led training and intervention team. The "Bruno Effect" resulted in a dramatic reduction in the number of physical attacks from an observed 5 fights per day (3 out of the 5 involved knives and cutting) to 1 per week. The violence rate returned immediately to its former level as soon as "Bruno" left the school. The dramatic violence reduction appears related to establishing an adult protective shield. Results stem from the unique personality of the adult protector, as well as a combination of the special role of the police and the outside intervention team. PMID- 9211043 TI - The violence of despair: consultation to a HeadStart program following the Los Angeles uprising of 1992. AB - This paper describes an acute trauma consultation to one Los Angeles HeadStart program following the severe civil disturbance in the spring of 1992. By self report, 7% (3/42) of the teachers had severe post-traumatic stress symptoms, and 29% (21/42) had moderate symptoms one month after the event. Teachers, comparing behavior following the disturbance to behavior two weeks before, reported statistically significant increases in the students' aggression and noise level, and decreases in "getting along" with peers and academic progress. Difficulties encountered in this type of community psychiatric consultation are discussed. PMID- 9211044 TI - Diversity in case management modalities: the Summit model. AB - Though ubiquitous in community mental health agencies, case management suffers from a lack of consensus regarding its definition, essential components, and appropriate application. Meaningful comparisons of various case management models await such a consensus. Global assessments of case management must be replaced by empirical studies of specific interventions with respect to the needs of specific populations. The authors describe a highly differentiated and prescriptive system of case management involving the application of more than one model of service delivery. Such a diversified and targeted system offers an opportunity to study the technology of case management in a more meaningful manner. PMID- 9211045 TI - Central role of vascular smooth muscle hyperreactivity in coronary hyperconstriction after balloon injury in miniature pigs. AB - BACKGROUND: Coronary constrictive responses to autacoids become augmented 1 week after balloon injury in our swine model. The present study aimed to elucidate the mechanisms of this effect. METHODS: In 12 hypercholesterolaemic miniature pigs, the coronary constrictive response to serotonin was examined angiographically 1 week after injury. After the angiographic study, organ chamber experiments using excised coronary artery were performed to clarify whether functional changes in endothelial cells or in vascular smooth muscle cells contributed to the hyperconstriction. RESULTS: The coronary constrictive response to serotonin in vivo was significantly greater at the previously injured site than at the non injured site. The degree of the hyperconstriction at the previously injured site exceeded that predicted from a geometric theory. Histological examination demonstrated that the previously injured site was almost covered with regenerated endothelial cells. In vitro studies demonstrated that serotonin caused significantly greater contraction in coronary artery strips, whether with or without endothelium, from the previously injured site than in those from the non injured site. Endothelium-dependent relaxation in response to serotonin was comparable at the injured and non-injured sites. CONCLUSIONS: These results suggest that the coronary hyperconstriction response to serotonin 1 week after injury results primarily from hyperreactivity of vascular smooth muscle. Whereas any contribution of endothelial dysfunction or the geometric effect may be minimal. PMID- 9211046 TI - Contrast echocardiography in coronary artery diseased patients: effect of systemic and pulmonary artery pressures on left heart opacification after intravenous injection of Albunex. AB - BACKGROUND: Contrast echocardiography is a useful tool for assessing repeatedly patients with coronary artery disease. Nevertheless, elevated pulmonary artery and systemic blood pressures likely to be associated with cardiac ischemia may limit the left ventricular opacification (LVO) because of the microspheres' sensitivity to pressure. OBJECTIVE: To determine the effects of systemic and pulmonary artery blood pressures on LVO. METHODS: We performed 55 intravenous injections (0.08 and 0.22 ml/kg) of a new transpulmonary contrast agent (Albunex), during two separated exposures, into 20 cardiac ischemic patients while monitoring invasively their cardiac indexes, and intracardiac, systemic, and pulmonary artery blood pressures. LVO was graded qualitatively from faint to full. RESULTS: A logistic model with the grade of LVO as the dependent variable and a selection from among the dose, exposure, right and left atrial blood pressures, systolic systemic and pulmonary artery blood pressures (ranges 94-208 and 14-45 mmHg, respectively), cardiac index, stroke index, and pulmonary and systemic vascular resistances as the explanatory variables demonstrated that increasing the dose gives an increasing probability of LVO (P = 0.02) and that increasing the pulmonary artery pressure reduces that probability (P = 0.006). A decreased cardiac index tended also to be associated with decreased LVO. The systemic blood pressure and the pulmonary and systemic vascular resistances had no statistically significant effect on the grade of LVO. CONCLUSIONS: LVO after intravenous administration of Albunex is dose-dependent and limited by an elevated pulmonary artery pressure. These data suggest that one should use higher doses for cardiac ischemic patients with elevated pulmonary artery pressures and that use of Albunex has the potential to detect pulmonary hypertension in patients. PMID- 9211047 TI - Comparative in-vitro validation of eight first- and second-generation quantitative coronary angiography systems. AB - BACKGROUND: It is known that first-generation quantitative coronary angiography (QCA) systems overestimate small vessel sizes owing to the point-spread function of the respective X-ray imaging chain. With second-generation systems new algorithms were introduced to correct for this source of error. OBJECTIVE: To evaluate the efficiency of the modified contour detection algorithms. METHODS: Six second-generation QCA systems (CMS, QANSAD, AWOS, CAAS II, Cardio 500, and Angioimage) were validated and compared with first-generation systems (CAAS and ARTREK). By using an arterial phantom consisting of stenotic and nonstenotic glass tubes (of diameters 0.5-5.0 mm) the accuracy and precision of each analysis system, as well as their additional accuracy and precision values for phantom diameters < or = 1.0 mm were determined. RESULTS: All systems had high accuracy and precision values, but first-generation systems overestimated small vessel diameters. With second-generation systems a significantly improved accuracy in the submillimeter range (an accuracy within +/-0.028 mm) was obtained. This improvement was accompanied by a moderate reduction in precision in the submillimeter range. CONCLUSION: The new algorithms of the second-generation QCA systems allow accurate and reliable measurements of small coronary dimensions and, therefore, precise analysis of coronary stenoses of moderate-to-high grade seems feasible with the improved accuracy of the new systems. PMID- 9211048 TI - The influence of age on the results of reoperative coronary artery bypass grafting. AB - BACKGROUND: With a steady increase in the number of elderly patients requiring coronary artery bypass grafting (CABG), a larger portion of elderly patients will also become candidates for reoperative CABG. Scepticism still exists as to whether this operation is justified in older patients. The purpose of this study was to examine the effect of increasing age on the outcome after reoperative CABG. METHODS: Between January 1, 1990 and June 30, 1996 563 patients underwent isolated reoperative CABG, and were included in this retrospective analysis. Patients who had combined procedures were excluded. The patients were divided by age into two groups: those aged 69 years or less (n = 507), and those older than 70 years (n = 56). Hospital mortality and morbidity for each group was compared. Medium-term survival for each group was compared with that of their age-matched population derived from Swiss life tables. RESULTS: The patients aged 70 years and older had a higher New York Health Association functional class, and more patients had unstable angina requiring urgent surgery than did the younger patients. The elderly also showed an over-representation of diabetes and multifocal vascular disease (generalized arteriosclerotic disease), and there was a higher number of patients with triple-vessel disease and left stenosis (> or = 70%) in this group. Patients aged 70 years and older received fewer distal anastomoses (3.0 versus 3.6; P < 0.01), and had a longer cardiopulmonary bypass time compared with the younger patients, but the ischemia time was similar in both groups. Hospital mortality was higher in patients older than 70 years (7.1 versus 17.9%). There was an increased frequency of postoperative low cardiac output and a higher incidence of gastrointestinal complications and transient renal failure amongst the elderly patients (> or = 70 years). Despite a higher hospital mortality rate and slightly increased morbidity the 5-year survival was excellent, and comparable with the age-matched population in both groups [89.6% (< 70 years) and 76.2% (> or = 70 years)]. The cardiac event-free survival was 79.8% (< 70 years), and 69.9% (> or = 70 years) after 5 years. CONCLUSION: An acceptable early mortality and long-term survival together with good functional long-term results support the justification of reoperative CABG in older patients, at least up to the age of 80 years. PMID- 9211049 TI - Beneficial effect of nisoldipine in repeated coronary reperfusion. AB - BACKGROUND: We have demonstrated that when repeated reperfusion is performed after reocclusion, there is a decrease in the amount of myocardial salvage, despite early reperfusion. It is unknown whether this deleterious effect of repeated reperfusion can be antagonized. Therefore, we studied the effect of nisoldipine (a dihydropyridine calcium antagonist) on infarct size, using a repeated-reperfusion model. METHODS: The left anterior descending coronary artery was occluded in anaesthetized dogs. Thirty minutes after occlusion, dogs were allocated randomly to either the treatment group (n = 6; 6 micrograms/kg per min nisoldipine infused intravenously throughout the experiment) or the control group (n = 8; saline). Occlusion was maintained for 2 h, followed by 1 h of reperfusion, then 1 h of reocclusion and 2 h of second reperfusion. An in vivo area at risk was determined by gentian-violet staining, and infarct size was defined and quantitated by triphenyl-tetrazolium-chloride staining. RESULTS: Haemodynamic measurements were similar in both groups. Mass of necrosis/mass at risk was significantly smaller in the nisoldipine group (33.3 +/- 5.8%, mean +/- SEM) compared with controls (46.8 +/- 4.8%; P < 0.05). CONCLUSION: Treatment with nisoldipine induces a beneficial effect by reduction of infarct size in repeated coronary reperfusion. PMID- 9211050 TI - Single-dose intramuscular administration of sustained-release Angiopeptin reduces neointimal hyperplasia in a porcine coronary in-stent restenosis model. AB - BACKGROUND: In-stent restenosis results primarily from neointimal hyperplasia. In a previous study we showed that continuous subcutaneous Angiopeptin infusion for 1 week significantly reduces neointimal hyperplasia in a porcine coronary overstretch in-stent restenosis model. The present study evaluated the relative efficacy of immediate-release and sustained-release Angiopeptin in the same model. METHODS: Thirty pigs (n = 10 in each group) were randomly assigned to three groups: controls receiving no Angiopeptin (Group 1); a sustained-release treatment group receiving one time intramuscular administration of 20 mg of Angiopeptin (Group 2); and a systemic treatment group receiving continuous Angiopeptin over a 1-week period via a subcutaneous osmotic pump (200 micrograms/kg total dose) (Group 3). One oversized Palmaz-Schatz stent (mean stent/artery = 1.25) was subsequently implanted in the left anterior descending coronary artery. The degree of neointimal reaction was evaluated 4 weeks later by angiography (maximal per cent diameter stenosis) and histology (maximal neointimal area corrected for injury score). RESULTS: A trend towards a reduction in diameter stenosis was observed by angiography, despite a similar degree of injury (25 +/- 17% in Group 1, 13 +/- 8% in Group 2, and 14 +/- 9% in Group 3; P = 0.072 by ANOVA). Histology demonstrated that both Angiopeptin treatment strategies significantly reduced in-stent neointimal area compared with the control group (1.65 +/- 0.97 mm2 in Group 1 versus 0.93 +/- 0.41 mm2 in Group 2 versus 0.85 +/- 0.28 mm2 in Group 3; P = 0.016 by ANOVA). Measurement of plasma Angiopeptin levels revealed comparable levels in both treatment groups, which persisted for up to 2 weeks. CONCLUSIONS: This study shows that single-dose intramuscular administration of sustained-release Angiopeptin reduces in-stent restenosis as effectively as the prolonged systemic treatment requiring a subcutaneous pump. Thus, a practical, effective, pharmacologic therapy for preventing in-stent restenosis may be available and should be evaluated in patients. PMID- 9211052 TI - How does the coagulation system really work and why should we care? PMID- 9211051 TI - Direct thrombin inhibitors in cardiovascular disease. AB - Heparin, the most widely used antithrombin, suffers several limitations, including high inter-individual variability of anticoagulant response, a nonlinear dose-response curve, inability to inactivate clot-bound thrombin, a requirement for endogenous cofactors and inactivation by platelet factor 4 and heparinase. These shortcomings may explain its suboptimal efficacy and safety in the prevention of arterial vessel occlusion. Heparin's drawbacks may be overcome by direct thrombin inhibitors. The development of these specific antithrombins has been a major therapeutic goal of the past decade. The high expectations generated by the use of these compounds in experimental models of arterial thrombosis appeared to be confirmed by the initial phase I and II clinical studies. However, large phase III trials have been highly discouraging: three trials with hirudin have been interrupted as a result of a high incidence of serious adverse events. Two of these trials were subsequently restarted at lower doses and did not support an incremental efficacy of hirudin over heparin. Two trials in the setting of angioplasty (one with hirudin and one with hirulog) have also failed to demonstrate the superiority of these compounds over heparin. Is this the result of a very narrow therapeutic range of these agents or the consequence of poor design of the phase II studies leading to the selection of inappropriate doses for the comparative efficacy trials? This review focuses on the clinical development of two specific antithrombins: hirudin and hirulog. The experimental pharmacology and human studies of Argatroban are discussed in a review by Fitzgerald and Murphy. PMID- 9211053 TI - Bibliography current world literature. PMID- 9211054 TI - Construction and practice of medical responsibility: dilemmas and narratives from geriatrics. AB - A narrative approach is employed in this article about dilemmas and physician reasoning in geriatric medicine in order to explore the moral-medical worlds of urban American physicians. Reconstructed dilemmas, in the form of stories told by 51 doctors, are analyzed as cultural documents of both clinical-moral knowledge and practice and the physician as moral actor. Discussion focuses on ways in which responsibility is constituted and enacted through a particular language of clinical action. This analysis opens the subject of bioethics to a range of infrequently discussed issues that physicians cite as deeply troubling and contributes to a broadening of anthropological approaches useful in the study of bioethics. PMID- 9211055 TI - Egg phobia in retirement homes: health risk perceptions among elderly Chinese. AB - Studies pertaining to health promoting behavior in daily life have received scant attention among medical anthropologists. The present study addresses this issue by means of an empirical analysis of perceptions and behaviors concerning a daily food item -eggs. Data were collected via in-depth interviews as well as participant observation in four retirement homes-two in Los Angeles and two in Taipei, Taiwan. The results reveal that practices of egg-restriction are pervasive throughout the four homes. Cholesterol has become a commonly-discussed issue in the daily lives of the 203 residents interviewed, and many of them were found to be preoccupied with the risk involved in excess consumption (especially of egg yolks) and increased serum cholesterol levels. Four forces: health professionals, family members, peer groups and mass media play important roles in constructing egg-consumption behaviors among the elderly subjects. The cognitive, psychological and behavioral impact of health information on elderly subjects has been discussed herein. It may be argued that the 'egg issue' reflects a shift in previous health paradigm thinking due to the biomedicalization of health promotion among Chinese elderly. PMID- 9211056 TI - Alternative psychotherapeutic practice among middle class Americans: I: Case studies and follow-up. AB - Historically, alternative psychotherapeutic procedures have florished worldwide in various inspirational, spiritualistic and shamanistic versions. However, few investigations have described these interventions in detail, or followed the clinical outcomes longitudinally. This present report is an ethnographic and clinical description of one particular American practitioner's alternative method as it is used with middle class clientele in several regions of the USA. Two case studies are presented in detail while nine individual cases are qualitatively reviewed with one month and one year treatment outcomes reported. A condensed ethnobiographical study of the practitioner's personal and professional life is included. PMID- 9211057 TI - Illness and morality in the Mombasa Swahili community: a metaphorical model in an Islamic culture. AB - The Swahili of Mombasa are Muslims, part of an ethnic group whose forebears founded East Africa's pre-colonial cities. Their cosmopolitan culture, in common with the cultures of some other, mainly Muslim, groups elsewhere includes a system of beliefs concerning the body's functioning and illness as the result of relations among the body's four humours. The Swahili version of this system and its use in curing is described and briefly compared to several others. Despite the availability of a variety of other systems, including the biomedical employed in the much patronized cost free treatment at government hospitals, the humoural system of beliefs is almost universally held. An hypothesis seeking to explain this suggests that because the Swahili schema is conceptualized by what Lakoff calls a "metaphorical model" emphasizing balance which is also used to conceptualize the values applying to key social relationships, the two schemata support one another, presumably by making the knowledge and emotions applying to one available in the other. PMID- 9211058 TI - Treatment of an Indian woman with major depression by a Latina therapist: cultural formulation. PMID- 9211059 TI - New insights into the regulation of lipoprotein metabolism: studies in procaryocytes, eukaryocytes, rodents, pigs, and people. PMID- 9211060 TI - Recent advances in elucidating the role of the microsomal triglyceride transfer protein in apolipoprotein B lipoprotein assembly. AB - The microsomal triglyceride transfer protein is necessary for the assembly and secretion of lipoproteins containing apolipoprotein B. During the past year, significant progress has been made towards understanding the role of microsomal triglyceride transfer protein in lipoprotein assembly at both a cellular and molecular level. Studies carried out in a variety of heterologous expression systems, as well as the use of microsomal triglyceride transfer protein inhibitors in hepatoma cell lines, have been critical to this progress. It has been shown that microsomal triglyceride transfer protein plays a key role in the early stages of lipoprotein assembly, most likely by transferring lipid to nascent apolipoprotein B as it enters the lumen of the endoplasmic reticulum. The evidence indicates that microsomal triglyceride transfer protein does not play a major role in addition of bulk core lipid in the late stages of apolipoprotein B48 lipoprotein assembly. Thus, microsomal triglyceride transfer protein appears to control the number of apolipoprotein B lipoprotein particles secreted but not the lipid composition. PMID- 9211061 TI - 3-Hydroxy-3-methylglutaryl coenzyme A reductase inhibitors and hepatic apolipoprotein B secretion. AB - Important advances in our understanding of the regulation of hepatic apolipoprotein B secretion have been made in the past year. A diverse group of studies have provided evidence that the inhibition of cholesterol synthesis by 3 hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors decreases the hepatic assembly and secretion of apolipoprotein B-containing lipoproteins. Apolipoprotein B kinetic studies performed in animals and human individuals indicate that inhibition of VLDL-apolipoprotein B secretion is an important mechanism whereby reductase inhibitors decrease plasma concentrations of these lipoproteins. Studies in cultured hepatocytes and in-vivo animal models have provided insights into how reduction of cholesterol synthesis decreases apolipoprotein B secretion. A decrease in hepatic acyl-coenzyme A: cholesterol acyltransferase activity, secondary to reduced microsomal cholesterol concentrations, has been implicated. PMID- 9211062 TI - Fatty acid regulation of very low density lipoprotein production. AB - The topic covered in this review is the regulation of hepatic VLDL production by fatty acids, with emphasis on the role of plasma free fatty acids. Hepatic VLDL production is primarily substrate driven, the most important regulatory substrate being fatty acids. Fatty acids may be derived from at least four sources: (1) de novo lipogenesis, (2) cytoplasmic triglyceride stores, (3) fatty acids derived from lipoproteins taken up directly by the liver, or (4) exogenous fatty acids (plasma free fatty acids). Although the total flux of fatty acids reaching hepatocytes plays an important regulatory role in VLDL synthesis, it is the nutritional and hormonal state of the organism that ultimately determines the rate of VLDL secretion. Nutritional and hormonal factors will determine whether intracellular fatty acids are channelled into oxidative, storage or secretory pathways. Conditions associated with both elevated free fatty acid flux to the liver and elevated de-novo lipogenesis, such as occurs in hyperinsulinemic insulin-resistant states, have hepatocytes primed to channel fatty acids into secretory pathways, with consequent high rates of VLDL secretion. Insulin resistant states are associated not only with the release of larger quantities of free fatty acids from the increased mass of circulating lipoproteins, particularly in the postprandial state, but also with reduced free fatty acid uptake and esterification by peripheral tissues. Thus a vicious cycle is set up in insulin-resistant states involving free fatty acids and hypertriglyceridemia. PMID- 9211063 TI - Apolipoprotein C-III gene variation and dyslipidaemia. AB - Within the apolipoprotein A-I-C-III-A-IV gene cluster, the Ssti polymorphism in the 3' untranslated region of the apolipoprotein C-III gene is the variant site most consistently and strongly associated with raised plasma triglyceride levels and coronary artery disease. To date the molecular mechanisms that underlie this effect have not been fully identified and variation within the insulin-responsive element of the apolipoprotein C-III proximal promoter cannot entirely explain the Ssti effect. PMID- 9211064 TI - Peroxisome proliferator-activated receptors, orphans with ligands and functions. AB - The three peroxisome proliferator-activated receptors (PPARs), PPAR alpha, delta and gamma, form a subfamily of the nuclear hormone receptor gene family. PPAR alpha has been shown to bind and be activated by leukotriene B4 and fibrates, whereas prostaglandin J2 derivatives and the antidiabetic thiazolidinediones, respectively, are natural and synthetic ligands for PPAR gamma. The availability of ligands and activators for PPAR alpha and PPAR gamma allowed an initial assessment of their respective functions. PPAR alpha and PPAR gamma are shown to function as important regulators in lipid and glucose metabolism, adipocyte differentiation, inflammatory response and energy homeostasis. PPAR alpha seems to mediate its pleiotropic effects mainly through the stimulation of oxidation of lipids, whereas PPAR gamma is a key mediator of lipid storage. The next few years will be very exciting as additional studies will refine our current knowledge about PPAR alpha and PPAR gamma and may reveal a ligand and role for the lonesome orphan among the PPARs, PPAR delta. PMID- 9211065 TI - Molecular aspects of intracellular sterol esterification: the acyl coenzyme A: cholesterol acyltransferase reaction. AB - The conversion of cholesterol to an esterified storage form by the enzyme acyl coenzyme A: cholesterol acyltransferase, is a critical component of sterol and membrane homeostasis and represents a significant step in atherogenesis. The isolation in 1993 of the first molecular probe of acyl-coenzyme A: cholesterol acyltransferase provided a key to understanding this reaction and its regulation. The sequence is apparently ubiquitous in eukaryotes, often with multiple sequence homologs within an organism. Presented here is a review of the known molecular events that lead to intracellular sterol esterification. PMID- 9211066 TI - Lipoprotein glycation and its metabolic consequences. AB - Glycation of lipoproteins is implicated in the development of the macro- and microvascular complications of diabetes, atherosclerosis in general, and other disease processes including aging. Enhanced glycation may have direct effects, and may also amplify the effects of oxidative stress on lipoproteins. Most studies have examined the effects of glycation of LDL, particularly with respect to its atherogenicity. Other lipoproteins are more difficult to study because their several apolipoproteins, being of varying age, are not uniformly exposed to glucose. Inhibition of the combined stresses of glycation and oxidation towards lipoproteins may have beneficial effects on health. PMID- 9211067 TI - Scavenger receptor BI--a cell surface receptor for high density lipoprotein. AB - The receptor-mediated transfer of lipids between cells and lipoproteins plays an important role in lipoprotein metabolism and cardiovascular disease. Although there have been many valuable studies of HDL binding to tissues, cells and membranes, and of the potential role of such binding in the transport of lipids between HDL and cells, much less is known about HDL receptors than about receptors for other lipoproteins (e.g. LDL, chylomicrons, vitellogenin). Here we review recent studies of the class B, type I scavenger receptor, which appears to be a physiologically relevant, cell surface HDL receptor that mediates the selective uptake of lipids by cells. PMID- 9211069 TI - Lipid metabolism. PMID- 9211068 TI - Genetic determinants of high density lipoprotein levels. AB - Low serum concentrations of HDL-cholesterol are well established markers for a high risk to develop coronary artery disease. The absence of coronary artery disease symptoms from some individuals with familial HDL deficiency demonstrates the presence of heterogeneity in this relation and points to an important role of etiologic and modifying factors. PMID- 9211070 TI - Nutrition. PMID- 9211071 TI - Genetics and molecular biology. PMID- 9211072 TI - Lipid metabolism. PMID- 9211073 TI - Hyperlipidaemia and cardiovascular disease. PMID- 9211074 TI - Atherosclerosis. PMID- 9211075 TI - Therapy and clinical trials. PMID- 9211076 TI - Primary prevention of coronary heart disease. What has WOSCOPS told us and what questions remain? West Of Scotland Coronary Prevention Study. AB - Coronary heart disease remains the major cause of death and morbidity in developed countries. As a consequence, its prevention constitutes a significant public health challenge. In recent times, our understanding of this disease process has expanded and many of the factors that influence its expression have been elucidated. In addition, a number of trials of diet and lipid-lowering drugs have been performed in an effort to tackle this problem. These studies demonstrate that when lipid levels are favourably altered, cardiovascular events are reduced without adverse effect. The rate at which event outcomes diverge between treated and untreated patients depends on the degree of atherosclerosis manifestation prior to treatment and the aggressiveness of the lipid altering strategy. Nonetheless, to date, the residual risk of cardiovascular events is still unacceptably high despite even the most potent lipid-lowering treatments used in these trials. In order to minimise the risk of future events, earlier intervention and a greater change in LDL and HDL cholesterol levels are needed in conjunction with other risk factor modifications. PMID- 9211078 TI - Recognition and management of myositis. AB - Polymyositis and dermatomyositis are severe inflammatory muscle disorders of unknown cause, with possible life-threatening complications. Prognosis and response to therapy may be predicted not only from the clinical and pathological diagnostic group to which a patient belongs, but also from the patient's myositis specific antibody status, extraskeletal muscle involvement, and the interval between onset of muscle weakness and the start of the treatment. Corticosteroids are the mainstay of treatment, providing recovery of normal muscular function in about 60% of patients. However, adequate dose, duration of therapy and gradual tapering of corticosteroids are required to produce favourable biochemical and clinical outcomes. In patients refractory to or intolerant of corticosteroids, additional therapy, often involving an immunosuppressive agent or intravenous immunoglobulin, is required. The potential roles of plasmapheresis and total body irradiation must be balanced by their inconclusive efficacies and adverse effects. PMID- 9211079 TI - Recognition and management of bacterial arthritis. AB - Bacterial arthritis is a bacterial infection of the joint. Apart from the classical gonococcal arthritis, nongonococcal arthritides include specific forms such as mycobacterial or Borrelia burgdorferi arthritis. Almost any bacterium can cause arthritis, provided that the route of penetration and the host response are suitable. Weakening of the host's immune competence, pre-existing joint damage and invasive diagnostic or therapeutic procedures are the main risk factors for bacterial arthritis. Gram-positive cocci are the species most frequently involved. The pathogenesis of bacterial damage includes release of toxins, cell production of cytokines and autoimmune reactions to specific antigens. The diagnosis can be suspected clinically put must be confirmed by culture of the synovial fluid, a test which can be complemented by scintigraphy. Amplification of bacterial DNA by polymerase chain reaction is a new procedure that could become an important tool for quick diagnosis. Treatment is based on joint drainage and antibiotics, which should be started as soon as the diagnosis is suspected. Corollary strategies under investigation include corticosteroids to prevent joint damage, monoclonal antibodies to arthritogenic peptides of bacteria or to surface markets of host lymphocytes, and modulators of synovial fluid cytokines. PMID- 9211080 TI - Chemotherapeutic induction of labour. A rational approach. AB - Induction of labour, defined as stimulation of uterine contractions before the spontaneous onset of labour, is indicated when the condition of the mother or fetus precludes awaiting the onset of spontaneous labour. In current medical practice, induction of labour comprises 2 phases: cervical priming and induction of contractions. Although numerous agents have been used for cervical priming, the current standard of care is the use of intracervical or intravaginal prostaglandin E2. The only drug currently used for induction of contractions is intravenous oxytocin. While many protocols are deemed acceptable, when required, the use of cervical priming, amniotomy and intravenous oxytocin are advocated. Utilising this approach, rapid delivery can be obtained in the majority of women. PMID- 9211081 TI - The role of advanced generation macrolides in the prophylaxis and treatment of Mycobacterium avium complex (MAC) infections. AB - Since the start of the acquired immunodeficiency syndrome (AIDS) epidemic, the role of Mycobacterium avium complex (MAC) as an opportunistic pathogen in advanced AIDS patients has become more and more clear. Once identified in an advanced AIDS patient it is possible to find evidence that the MAC organism and infection is not only present in the pulmonary tree, but has also disseminated to a wide variety of body organs. Treatment of MAC or disseminated MAC (DMAC) infections has historically been very difficult due to the inherent resistance of the MAC pathogen to most standard antimycobacterial agents. This has resulted in the development of new agents for the prevention of DMAC infection as well as combinations of both new and standard agents for its treatment. Three drugs are currently approved for single-agent DMAC prophylaxis, including rifabutin, azithromycin and clarithromycin. Combinations of agents for DMAC treatment are highly variable in content but most experts agree that all combinations should contain one of the advanced generation macrolides (azithromycin or clarithromycin). Both of these agents have favourable intracellular pharmacokinetics and pharmacodynamics which maximise their effects against this mostly intracellular pathogen. Due to the paucity of comparative data, no one macrolide can be recommended over the other. However, the expected increase in compliance, lower weekly and annual costs, and lack of any drug interactions may make azithromycin a preferable choice, but this should be decided on a case-by case basis. PMID- 9211082 TI - Nelfinavir. AB - Nelfinavir is a protease inhibitor which shows good inhibitory activity against HIV-1. The pattern of HIV-1 resistance to nelfinavir is different from that seen with other protease inhibitors. In healthy male volunteers, administration of single 400 and 800mg doses of nelfinavir with food resulted in area under the plasma concentration-time curve values that were 27 to 50% higher than those achieved in fasted volunteers who received the same doses of the drug. Reductions in plasma HIV RNA to below detectable levels (detection limit 500 copies/ml) were achieved in some patients (number not reported) after 28 days' treatment with nelfinavir 500, 600 or 750 mg twice daily, or 500, 750 or 1000 mg 3 times daily. Combination therapy with nelfinavir and stavudine produced greater reductions in plasma HIV RNA levels that stavudine monotherapy in patients who had not previously received stavudine treatment; mean increases in CD4+ cell counts were also greater in the combination treatment group than in monotherapy recipients. Plasma HIV RNA decreased to below detectable levels in 11 of 12 patients with early onset HIV infection who received a triple regimen of nelfinavir, zidovudine and lamivudine for 16 weeks. PMID- 9211083 TI - Troglitazone. AB - Troglitazone decreases insulin resistance (improves insulin sensitivity), which results in reduced plasma glucose and insulin levels in patients with non-insulin dependent diabetes mellitus (NIDDM). Risk factors for cardiovascular disease such as elevated proinsulin and triglyceride levels are also reduced by troglitazone. In clinical trials, troglitazone 200 to 800 mg daily (alone or in combination with other oral antidiabetic agents or insulin) reduced plasma or serum glucose levels and glycosylated haemoglobin compared with both baseline and placebo in patients with NIDDM refractory to other oral antidiabetic agents (usually sulphonylureas). Troglitazone was generally well tolerated in clinical trials. In patients in the US, the incidence of adverse events in troglitazone recipients was similar to that in placebo recipients. PMID- 9211084 TI - Fosinopril. A review of its pharmacology and clinical efficacy in the management of heart failure. AB - Fosinopril is the prodrug of the active diacid ACE inhibitor fosinoprilat. In patients with heart failure, fosinopril reduces pulmonary capillary wedge pressure, mean arterial blood pressure, mean right atrial pressure and heart rate, and increases stroke volume index and cardiac index. The drug has compensatory dual elimination routes via renal and hepatic systems and accumulates to a lesser extent than enalapril and lisinopril in patients with chronic renal insufficiency with or without heart failure. Comparative studies of 3 or 6 months' duration with fosinopril 10 to 40 mg/day have demonstrated clinical efficacy significantly superior to that of placebo in patients with heart failure [mostly New York Heart Association (NYHA) functional class II or III]. Fosinopril treatment consistently increased exercise duration and improved heart failure symptoms in these patients. Significantly fewer fosinopril than placebo recipients withdrew or were hospitalised because of worsening heart failure. Additionally, significantly more fosinopril than placebo recipients showed improvement, and fewer patients had deteriorated, in terms of NYHA functional class. Fosinopril and enalapril showed similar clinical efficacy over 6 and 12 months' treatment in patients with NYHA functional class II to IV heart failure. As yet, there are no data showing a mortality benefit with fosinopril. Fosinopril was well tolerated in clinical trials in patients with heart failure. Dizziness (11.9 vs 5.4% for placebo), cough (9.7 vs 5.1%) and hypotension (4.4 vs 0.8%) were the most commonly reported adverse events. In 6- or 12-month comparative studies, fosinopril therapy was associated with a lower incidence of dizziness and hypotension, but a higher incidence of vertigo, than enalapril therapy. 0.8% of patients discontinued the drug because of cough, which occurred to a similar extent with fosinopril and enalapril. Thus, based on available clinical evidence, fosinopril is an effective and well tolerated option for the management of patients with heart failure. Although clinical data are limited, fosinopril may be especially useful in patients with renal or hepatic impairment. PMID- 9211077 TI - Trends and future developments in the pharmacological treatment of acute ischaemic stroke. AB - Stroke stands as the third leading cause of death. It makes great demands on patients, who must not only survive the complications of the acute stages, but must cope then with the great physical and economic costs of long-term disabilities. Therefore, there is urgent need to establish generally useful regimens for the acute treatment of ischaemic stroke. Three treatment approaches are based upon pathophysiologic concepts derived from experimental work with focal cerebral ischaemia. These include pharmacologic strategies for arterial recanalisation, inhibition of inflammatory processes and neural protection. Focal cerebral ischaemia secondary to occlusion of a brain-supplying artery initiates neuronal and microvascular events, and the simultaneous processes of inflammation which further injure tissue. The use of plasminogen activators to mediate thrombus and lysis in the acute setting has been shown to be clinically beneficial. Further work with arterial reperfusion strategies is under way. Early clinical studies with polymorphonuclear leukocyte-dependent endothelial adhesion receptor antagonists are being completed, but a strategy has yet to emerge. A large effort examining the potential efficacy of agents which may stabilise or protect neurons from ischaemic injury has shown promise in experimental models, and has been translated into clinical trials. Experimental work, and limited clinical experience, have indicated that: (a) the time window for intervention is important in limiting ischaemic and inflammatory injury, and for reducing the risk of haemorrhagic transformation; (b) putative neuroprotective strategies may potentially elongate the time interval for treatment; and (c) limitations from the adverse effects of plasminogen activators and of agents which beneficially affect neuronal dysfunction during ischaemia must yet be overcome. This review surveys pharmacological approaches currently undergoing evaluation which provide the goal of establishing effective strategies for the treatment of patients with acute cerebral ischaemia. PMID- 9211085 TI - Cefpirome. A review of its antibacterial activity, pharmacokinetic properties and clinical efficacy in the treatment of severe nosocomial infections and febrile neutropenia. AB - Cefpirome is an injectable extended-spectrum or 'fourth generation' cephalosporin. Its antibacterial activity encompasses many of the pathogens involved in hospital-acquired infections such as Enterobacteriaceae, methicillin susceptible Staphylococcus aureus, coagulase-negative staphylococci and viridans group streptococci. Cefpirome also has in vitro activity against Streptococcus pneumoniae regardless of penicillin susceptibility. It is stable against most plasmid- and chromosome-mediated beta-lactamases, with the exception of the extended-spectrum plasmid-mediated SHV enzymes. Intravenous cefpirome 2g twice daily has shown clinical efficacy comparable to that of ceftazidime 2g 3 times daily in the treatment of hospitalised patients with moderate to severe infections. Clinical response and bacteriological eradication rates were similar in patients with severe pneumonia or septicaemia treated with either cefpirome or ceftazidime. Cefpirome appeared more effective than ceftazidime in the eradication of bacteria in patients with febrile neutropenia in 1 study; however, clinical response rates were similar in the 2 treatment groups. The tolerability of cefpirome appears similar to that of ceftazidime and other third generation cephalosporins, diarrhoea being the most frequently observed event. Thus, cefpirome is likely to be a valuable extended-spectrum agent for the treatment of severe infections. Cefpirome offers improved coverage against some Gram-positive pathogens and Enterobacteriaceae producing class I beta-lactamases compared with the third generation cephalosporins, although this has yet to be demonstrated in clinical trials. PMID- 9211088 TI - Nefazodone. Psychomotor and cognitive effects. PMID- 9211086 TI - Toremifene. A review of its pharmacological properties and clinical efficacy in the management of advanced breast cancer. AB - The triphenylethylene antiestrogen toremifene is a chlorinated derivative of the antiestrogen tamoxifen, an agent which has been widely and successfully used in the treatment of breast cancer. Clinical trials investigating the efficacy of toremifene as first-line endocrine therapy in postmenopausal women with advanced breast cancer (estrogen receptor status positive or unknown) have shown this drug to have similar antitumour activity to that of tamoxifen. In multicentre comparative trials, objective responses (complete and partial) occurred in 20 to 29% of patients treated with toremifene (60 to 240 mg/day) and in 19 to 37.5% of tamoxifen (20 or 40 mg/day) recipients. The duration of response, time to disease progression and median overall survival time were generally similar in both treatment groups. Toremifene is well tolerated. Most drug-related adverse effects are mild or moderate in severity and rarely necessitate discontinuation of therapy. The tolerability profile of toremifene is similar to that reported for tamoxifen, the most common adverse effects being hot flushes, sweating, nausea and/or vomiting, dizziness, oedema, and vaginal discharge and/or bleeding. Thus, toremifene provides an equally effective and well tolerated alternative to tamoxifen for the first-line endocrine therapy of postmenopausal advanced breast cancer. Preclinical studies showing toremifene to have a lower carcinogenic potential than tamoxifen indicate that toremifene may be a preferable agent for long term treatment regimens; however, these findings require confirmation in the clinical setting. PMID- 9211090 TI - [Prognosis for medically treated elderly patients with coronary artery disease: analysis by the Cox model]. AB - The prognostic importance of age among well-known prognostic factors such as extent of coronary artery lesions, cardiac function, and myocardial ischemia was evaluated in 147 elderly patients with coronary artery disease aged 65 years or older who underwent dipyridamole perfusion scintigraphy and coronary angiography. After excluding 32 patients who initially underwent percutaneous transluminal coronary angioplasty (PTCA) or coronary artery bypass grafting (CABG), 115 patients who were initially treated medically were analysed by the Cox model for cardiac events during a mean follow-up period of 29 +/- 22 months. Among the 114 patients who were available for follow-up, nine patients (7.9%) had cardiac events, including five cardiac deaths and four non-fatal cardiac events (requiring PTCA or CABG). When the 114 patients were divided into three age groups; 53 patients aged 65-69 years, 42 aged 70-74 years and 19 aged 75 years or older, the incidence of cardiac death was highest in those aged 75 years or older. Univariate analysis showed that age of 70 years or older (hazards ratio 15.15, p = 0.004), scintigraphic diffuse slow washout (hazards ratio 8.77, p = 0.002), and triple-vessel or left main trunk disease (hazards ratio 6.36, p = 0.05) were important prognostic factors. Multivariate analysis showed that scintigraphic diffuse slow washout (hazards ratio 6.33, p = 0.05), and triple vessel or left main trunk disease (hazards ratio 11.94, p = 0.05) were statistically significant as independent prognostic factors. However, when age of 70 years or older was included in the analysis, it showed higher hazards ratio (21.21, p = 0.03) than that of scintigraphic diffuse slow washout (7.36) or triple-vessel or left main trunk disease (5.30). Age of 70 years or older may be a significant prognostic factor in elderly patients with coronary artery disease which has an equivalent importance to the extent of coronary lesions. PMID- 9211089 TI - [Gene Polymorphism of 5, 10-methylenetetrahydrofolate reductase as a coronary risk factor]. AB - Hyperhomocysteinemia has been identified as a possible risk factor for coronary artery disease. The association of the alanine/valine (A/V) polymorphism of 5, 10 methylenetetrahydrofolate reductase (MTHFR), one of the key enzymes catalyzing re methylation of homocysteine, with coronary artery disease was examined in 362 Japanese males with a diagnosis of coronary artery disease confirmed with coronary angiography. The A/V polymorphism was analyzed with PCR followed by Hinf I digestion. The screening of 778 male volunteer controls revealed that the frequency of V allele in Japanese was 0.33, comparable to that in the French Canadian population. The VV genotype, which correlates with increased plasma homocysteine levels due to reduced activity and increased thermolability of this enzyme, was significantly more frequent in patients with coronary artery disease (15.7%, n = 362) than in controls (10.2%, n = 778; p = 0.0067). The association of the VV genotype with coronary artery disease was further increased in patients with > or = 99% stenotic lesion (p = 0.0010). In these patients, the frequency of the VV genotype was significantly higher in patients with triple-vessel disease (26%) than in patients with single- or double-vessel disease (15% and 14%, respectively). The fasting plasma homocysteine levels in VV subjects were higher than those in AV or AA subjects. The VV genotype of MTHFR associated with increased plasma homocysteine levels may represent an important genetic risk factor for coronary artery disease, especially with the occurrence of myocardial infarction. PMID- 9211087 TI - Carvedilol. A reappraisal of its pharmacological properties and therapeutic use in cardiovascular disorders. AB - Carvedilol competitively blocks beta 1, beta 2 and alpha 1 receptors. The drug lacks sympathomimetic activity and has vasodilating properties that are exerted primarily through alpha 1-blockade. Animal models indicate that carvedilol confers protection against myocardial necrosis, arrhythmia and cell damage caused by oxidising free radicals, and the drug has no adverse effects on plasma lipid profiles. Recent data have confirmed the antihypertensive efficacy of carvedilol in patients with mild to moderate essential hypertension. Carvedilol has similar efficacy to other beta-blocking agents, calcium antagonists, ACE inhibitors and hydrochlorothiazide. Carvedilol also improves exercise tolerance and ischaemic symptoms in patients with stable angina pectoris. Significant reductions in serious cardiac events after acute myocardial infarction and in frequency and severity of ischaemic events in patients with unstable angina have also been demonstrated. Interest in the use of carvedilol in patients with congestive heart failure (CHF) has culminated in the publication of a cumulative analysis of data from 1094 patients with mild to severe CHF who participated in the US Carvedilol Heart Failure Study Program (4 trials). After a median follow-up of 6.5 months, a significant overall reduction in mortality relative to placebo (3.2 vs 7.8%) was revealed in patients who had received carvedilol 6.25 to 50 mg twice daily (plus diuretics and ACE inhibitors). All-cause mortality, risk of hospitalisation for cardiovascular reasons and hospitalisation costs were also reduced significantly (by 65, 28% and 62%, respectively) in these trials. In addition, the Australia and New Zealand Heart Failure Research Collaborative Group showed a 26% reduction in the combined risk of death or hospitalisation with carvedilol 12.5 to 50 mg/day relative to placebo after a mean 19-month follow-up period in 415 patients with CHF (relative risk 0.74). Adverse events with carvedilol appear to be less frequent than with other beta-blocking agents, are dosage-related and are usually seen early in therapy. Events most commonly reported are related to the vasodilating (postural hypotension, dizziness and headaches) and the beta blocking (dyspnoea, bronchospasm, bradycardia, malaise and asthenia) properties of the drug. Carvedilol appears to date to have little effect on the incidence of worsening heart failure. Concomitant administration of carvedilol with some medications requires monitoring. Carvedilol is therefore likely to have a beneficial role in the management of controlled CHF, but further clinical studies are required to show the place of beta-adrenoceptor blocking therapy in general in this indication, and the position of carvedilol relative to other similar agents. Carvedilol is also confirmed as effective in the management of mild to moderate hypertension and ischaemic heart disease. PMID- 9211091 TI - [Usefulness of directional coronary atherectomy as a bail-out device for acute closure after coronary angioplasty]. AB - The usefulness of directional coronary atherectomy (DCA) as a bail-out device for acute closure (reclosure) after percutaneous transluminal coronary angioplasty (PTCA) was evaluated. PTCA was performed in 1,023 patients (182 with acute myocardial infarction) between January 1993 and January 1994 in our hospital. Thirty-one patients (11 with acute myocardial infarction) suffered acute closure (reclosure) after PTCA. In six patients (five with acute myocardial infarction), DCA was performed as a rescue treatment for acute closure (reclosure). In three of these patients, angioscopy was performed before DCA, which demonstrated intimal tear and some thrombi although coronary angiography showed no evidence of thrombus. Bail-out DCA was successful in all six patients without complications. DCA is useful as a bail-out device for acute closure (reclosure) after PTCA when the thrombus is not massive. PMID- 9211092 TI - [Assessment of auscultatory blood pressure measurements versus intra-arterial pressure in patients with atrial fibrillation]. AB - The accuracy of auscultatory blood pressure (BP) determination was assessed in patients with chronic atrial fibrillation by performing simultaneous auscultatory BP determination on the upper arm and a direct BP determination on the contralateral arm. The subjects were three hospitalized patients, aged from 52 to 75 years. A Teflon catheter was introduced into the radial artery which was connected to a pressure transducer, and a cuff was twisted around the contralateral upper arm in the supine position. Simultaneous recording of directly determined BP and cuff pressure enabled the comparison of direct BP with auscultatory BP. The appearance of the Korotkoff I sound (systolic BP) and V sound (diastolic BP) was marked on the cuff pressure curve. This maneuver was repeated five times in each patient. The method of Bland and Altman was employed to assess the agreement between auscultatory and direct determinations. The auscultatory method estimated BP with differences of -14.3 to +27.3 mmHg in systolic BP and -12.1 to +11.9 mmHg (+/-2SD) in diastolic BP compared with the direct method. The difference in systolic BP between the auscultatory and the direct methods was greater than that in diastolic BP. Thus, there are unacceptable differences in systolic BP between auscultatory and direct methods that can be attributed to BP fluctuations. The auscultatory method in diastolic BP is more accurate than that in systolic BP and may be more useful in the clinical setting. PMID- 9211093 TI - Myocardial contractility of the canine left ventricle during unsynchronized dual chamber pacing. AB - The influence of changes of left ventricular (LV) myocardial contractility on the decrease of cardiac output during atrial fibrillation was investigated in dogs using the slope (Ec) and the length intercept (Lo) of the LV end-systolic force length relationship. The hearts of nine healthy adult mongrel dogs were instrumented with ultrasonic crystals and a micromanometer, after which pharmacologic autonomic blockade was instituted. The LV diameter and pressure data were recorded during inferior vena caval occlusion. Hemodynamic parameters were measured during pacing from the right atrial appendage at a pacing rate 30 beat/min greater than the natural heart rate using a cardiac stimulator (atrial pacing), and during simultaneous pacing from the right atrial appendage and right ventricular apex at the same rate (unsynchronized dual chamber pacing). Cardiac hemodynamics in the absence of synchronized left atrial contraction were simulated by unsynchronized pacing. During atrial pacing, the cardiac output (1.68 +/- 0.25 vs 1.57 +/- 0.21 l/min, p < 0.005) and Ec (110.1 +/- 58.5 vs 81.8 +/- 30.8 g/cm, p < 0.05) were significantly greater than during normal sinus rhythm, whereas the stroke volume (12.4 +/- 2.4 vs 15.1 +/- 3.1 ml, p < 0.005) and LV end-diastolic volume (16.6 +/- 2.7 vs 19.5 +/- 3.4 ml, p < 0.005) were significantly smaller. Lo did not change during pacing. During unsynchronized dual chamber pacing, cardiac output (1.46 +/- 0.17 vs 1.68 +/- 0.25 l/min, p < 0.005), stroke volume (10.8 +/- 1.7 vs 12.4 +/- 2.4 ml, p < 0.005), and LV end diastolic volume (15.0 +/- 2.0 vs 16.6 +/- 2.7 ml, p < 0.05) were significantly smaller than during atrial pacing. However, Ec and Lo were similar during both types of pacing. These findings suggest that the decrease of cardiac output and stroke volume during atrial fibrillation is chiefly due to the decrease of LV end diastolic volume through loss of left atrial contraction, and is not due to a change of LV myocardial contractility. PMID- 9211094 TI - [Long-term beneficial effect of dual chamber pacing in a patient with hypertrophic obstructive cardiomyopathy]. AB - Dual chamber (DDD) pacing therapy was effective to reduce the left ventricular outflow tract pressure gradient for a long time in a patient with pharmacotherapy resistant hypertrophic obstructive cardiomyopathy. A 52-year-old man with pharmacotherapy-resistant pressure gradient was treated by a DDD pacemaker implantation, because the pressure gradient was proved to be reduced (94-->16 mmHg) by transient DDD pacing with an atrioventricular delay of 50 msec. Hemodynamics and ventricular wall thickness were serially observed after the implantation for 2 years. The pressure gradient progressively decreased during the pacing period, at 4 months and 2 years follow-up, (10-->2 mmHg) and during the sinus rhythm period (60-->25 mmHg), and left ventricular ejection fraction and end-diastolic volume index were increased. Although the ventricular wall thickness remained constant, the systolic anterior motion of the mitral valve and A/E were reduced during the pacing period in the echocardiography. During the acute effect of DDD pacing, the pressure gradient reduction seemed to be related to dilatation of the left ventricular outflow tract induced by a change of contraction modality of the intraventricular septum. Improved left ventricular diastolic function may contribute to the pressure gradient reduction during extended periods of pacing therapy. PMID- 9211095 TI - [Cardiovascular imaging in a month. A 64-year-old man complaining of orthopnea]. PMID- 9211096 TI - Diagnosis of paradoxical embolism. PMID- 9211097 TI - [Adherence to valve repair surgery: from the mitral to the aortic valve]. AB - Mitral valve repair for regurgitation has recently become more predictable based on its high reproducibility and excellent long-term durability. Since October 1992, mitral valve repair has been attempted in 123 patients with dominant regurgitation and achieved in 121 patients (98%). The hospital mortality was 1.7% and reoperation was carried out in five patients (4.1%). Follow-up study of mitral regurgitation by echocardiography showed that the regurgitation was trivial or none in 100 patients (88%), mild in 9 (8%) and moderate or over in 5 (4%). The indication of mitral valve repair is now being expanded to more complex lesions, ischemic papillary muscle rupture, active infective endocarditis and recurrent lesion. Furthermore, repair for insufficient aortic valve was begun in January 1994 and has been performed in 34 patients (87%) of 39 with aortic regurgitation. The hospital mortality was 2.9% and reoperation was needed in two patients (6%). The degree of aortic regurgitation after discharge was estimated as trivial or none in 23 patients (70%), mild in 7 (21%), and moderate in 1 (3%). The results of mitral valve repair support an aggressive approach toward mitral regurgitation with valve repair. Although aortic valve repair seems to be less satisfactory, a definite group of patients will benefit by preventing the need for a prosthetic valve. PMID- 9211098 TI - [Experimental study on the pathogenesis of mitral annular calcification: calcium deposits in mitral complex lesions induced by vagal stimulation in rabbits]. AB - Cervical vagal stimulation in rabbits frequently causes peculiar mitral complex lesions which are detected by deposition of colloidal carbon. The present study examined the effects of vagal stimulation on the calcium (Ca) contents in the mitral complex lesions. Anesthetized rabbits were forced into the supine position with electrocardiographic monitoring. The animals were divided into those with vagal stimulation (n = 37), and those without manipulation as controls (n = 25). Colloidal carbon (1 ml) was intravenously injected into the animals on the next day. All animals were sacrificed after 1 week. After perfusion of the heart by heparinized saline, Ca content per g myocardium in the mitral annulus, papillary muscle, free wall or apex in the left ventricle was measured by the atomic absorption method. Immediately after vagal stimulation, bigeminy due to premature ventricular contractions was observed in 76%, and systolic murmur was heard in 30%. Mitral complex lesions detected by carbon deposits were found in 73% of the rabbits with vagal stimulation. Ca content in the mitral annulus or papillary muscle was significantly greater than that in the free wall or apex (p < 0.001). Ca content in the mitral annulus was significantly greater in the rabbits with vagal stimulation than in the control group (p < 0.001). In the former group, Ca content in the mitral annulus in animals with mitral complex lesions was significantly greater than that in those without mitral complex lesions (p < 0.01). These results suggest that these mitral complex lesions might provide an experimental model of mitral annular calcification, and that the autonomic nervous system and arrhythmia might be involved in the mechanism of this calcification. PMID- 9211099 TI - [Myocardial adrenergic nerve activity in valvular diseases assessed by iodine-123 metaiodobenzylguanidine myocardial scintigraphy]. AB - Iodine-123-metaiodobenzylguanidine (MIBG) imaging was used to assess myocardial adrenergic nerve activity in patients with heart failure MIBG planar images were obtained in 94 patients. The uptake of MIBG, calculated as the heart-to mediastinum activity ratio in the immediate image (15 min), showed a significant decrease only in patients with severe heart failure due to cardiomyopathy, but was not changed in those with valvular diseases. Storage and release of MIBG, calculated as the percentage myocardial MIBG washout from 15 min to 4 hours after isotope injection, was substantially accelerated in both patients with cardiomyopathy and valvular diseases in proportion to the severity of heart failure. These data suggest that, in severe heart failure associated with cardiomyopathy, norepinephrine uptake is reduced. Also, myocardial adrenergic nerve activity is accelerated in proportion to the severity of heart failure independent of the underlying cause. MIBG images were analyzed in 20 patients with mitral stenosis with the same methods to clarify whether myocardial adrenergic nerve activity is different in patients with heart failure without left ventricular volume or pressure overload. Myocardial uptake of MIBG did not show any significant difference. The percentage myocardial MIBG washout was increased in patients with severe heart failure. The closest correlation was between myocardial washout and cardiac output. In heart failure due to mitral stenosis, myocardial adrenergic nerve activity is intensified. Decrease in cardiac output associated with mitral stenosis acts as a potent stimulus for this intensification. PMID- 9211100 TI - [Variations in mesenchymal cell activity between rheumatic and non-rheumatic valve disease]. AB - Deoxyribonucleic acid (DNA) synthesis of mesenchymal cells in the diseased valve tissue of patients with rheumatic and non-rheumatic valve diseases were compared. Surgically resected mitral valves from eight rheumatic and eight non-rheumatic patients in their 40s were examined immunohistochemically to estimate the activity of the various mesenchymal cells by cell cycle analysis, using monoclonal antibodies to cyclin E, A, B1, p53, and b cl-2 after routine tissue fixation, paraffin embedding and sectioning. Cyclin B1-positive fibrocytes and fibroblasts of the spongiosa and fibrosa were observed in all non-rheumatic cases, whereas some lymphocytes in the perivascular area were cyclin B1-, p53- and b cl-2-positive in rheumatic cases. The distinct qualitative difference in the mesenchymal cells of valve tissue between rheumatic and non-rheumatic etiologies suggests a different mode of valve pathology. PMID- 9211101 TI - [Hemorheological effects of autologous blood storage before surgery for cardiac valvular diseases]. AB - The hemorheological effects of autologous blood storage with or without the use of erythropoietin were examined before surgery for valvular disease. There was no rheological difference between patients with aortic (16 cases) or mitral (10 cases) valve disease. Before storage, the levels of hematocrit, whole blood viscosity, and especially coefficient of rheology, were lower (p < 0.05) in the blood stored with erythropoietin, but this difference disappeared after storage. The plasma viscosity of both groups did not change before and after storage. The viscosity of blood was equalized after the storage of blood, irrespective of the use of erythropoietin. PMID- 9211102 TI - [Transesophageal echocardiographic findings of cortical and perforating infarctions in patients with atrial fibrillation]. AB - The predictive value of transesophageal echocardiography was investigated for the risk stratification of atherothrombotic or embolic cerebral infarction in patients with atrial fibrillation. Left atrial spontaneous echo contrast, peak flow velocity in the left atrial appendage and generalized atherosclerosis as estimated by the intima-media wall thickness of the thoracic aorta were assessed by transesophageal echocardiography in consecutive patients with paroxysmal (n = 25) or chronic (n = 60) atrial fibrillation (mean [+/-SD] age; 63 +/- 11 years). All patients underwent brain computed tomography or magnetic resonance imaging to evaluate the presence or absence of cerebral infarction. The location of cerebral infarction was divided into two territories, the cortical branch (cortical infarction) and deep perforators (perforating infarction), to evaluate embolic and atherothrombotic cerebral infarction, respectively. Cortical and perforating infarctions were found in 42% and 16% of all patients, respectively. The grade of spontaneous echo contrast was higher in patients with cortical infarction than in those without cortical infarction. Patients with perforating infarction showed thicker aortic wall compared with patients without perforating infarction. Other parameters had no predictive value to differentiate perforating from cortical infarctions. Multiple regression analysis revealed that spontaneous echo contrast and age were independent predictors of embolic cortical infarction, whereas intima-media wall thickness of the aorta and hypertension were useful in predicting the risk of atherothrombotic perforating infarction. Transesophageal echocardiography is useful for predicting embolic cortical infarction and atherothrombotic perforating infarction. PMID- 9211103 TI - [Mitral valve remodeling using valvuloplasty, chordoplasty and ring annuloplasty]. AB - Degenerative mitral valve disease is a major cause of mitral regurgitation and mitral valve repair has acquired greater importance as a surgical treatment of mitral regurgitation. Since 1991 we have used mitral valve repair to remodel the mitral valve leaflet, chordae tendineae and annulus. The final aim of our mitral valve remodeling technique is to correct the coaptation line of both leaflets. Forty-eight patients who underwent mitral valve repair were analyzed to evaluate the effect of the mitral valve reconstructive technique. We compared the left ventricular function and mitral valvular function by echocardiography and also the left ventricular pressure volume area, which reflects the oxygen consumption of the myocardium, before and after operation. The actuarial survival from all deaths, including one hospital death and one noncardiac death, was 95.8% at 60 months. Three patients have required reoperation because of recurrent regurgitation. Freedom from reoperation at 5 years was 93.8%. Left ventricular end-diastolic (122.3 +/- 45.8-->66.4 +/- 33.8 ml/ m2) and end-systolic volume indexes (39.6 +/- 19.5-->30.1 +/- 18.9 ml/m2) significantly decreased after mitral valve repair, and left ventricular volume overload was markedly reduced. Left ventricular ejection fraction and fractional shortening, which were apparently good before operation, were within the normal range after operation. Left ventricular inflow velocity and mitral pressure gradient were unchanged before and after repair. Mitral valve orifice area was 3.20 +/- 1.08 cm2 and did not show signs of mitral stenosis after repair. The reduction of left ventricular pressure volume area after repair was 53.6 +/- 12.4%, showing marked reduction of the oxygen consumption of the myocardium after successful mitral repair. Left ventricular and mitral valvular function is well preserved after mitral valve remodeling. Mitral valvular function after mitral repair exceeds that of the mitral valve replacement. Mitral valve remodeling procedures are very useful for patients with mitral regurgitation due to mitral valve prolapse. PMID- 9211104 TI - [Right atrial thrombus recognized 18 years after tricuspid valve replacement: a case report]. AB - A 53-year-old man, who had undergone tricuspid valve replacement with Hancock valve and direct closure of a ventricular septal defect when aged 34 years, was admitted with signs of right heart failure. Two-dimensional echocardiography showed bioprosthetic tricuspid valve malfunction with right atrial thrombus. He was treated by tricuspid valve replacement using a Hancock II valve and removal of the right atrial thrombus with remarkable improvement. Transesophageal echocardiography was the most useful method for recognizing the presence of right atrial thrombus and assessing its actual or potential hemodynamic effects. PMID- 9211105 TI - [Surgically treated unruptured sinus of Valsalva aneurysm: a case report]. AB - A 73-year-old man was admitted because of progressive dyspnea. Echocardiography revealed a markedly dilated right sinus of Valsalva and severe aortic regurgitation. Coaptation loss of the aortic valve leaflets was thought to be the cause of regurgitation and congestive heart failure, a rare complication of the unruptured sinus of Valsalva aneurysm. Surgical treatment should be considered to prevent life-threatening heart failure and rupture of the aneurysm even in patients without overt heart failure. PMID- 9211106 TI - [Aortic valve replacement with stentless porcine aortic valve]. AB - The Toronto stentless porcine valve (TSPV) is a stentless porcine aortic valve, preserved with glutaraldehyde solution, which has been designed to maintain natural laminar blood flow and improve hemodynamic performance and enhance durability. Aortic valve replacement with TSPV was performed in seven patients. The reason for this indication was advanced age (65 years or more) in six patients and connective tissue disease in another patient. One patient required reoperation because of prosthetic valve endocarditis 4 months after implantation. There were no other valve-related complications. Echocardiographic examination of prosthetic valve function one month after surgery showed no aortic regurgitation in any patient. Transprosthetic gradient was 15.1 +/- 5.0 mmHg. Technical know how is required for the implantation. The hemodynamic characteristics of the TSPV were excellent. Further follow-up is required to determine durability, but further extensive indication of this valve is expected as a bioprosthesis. PMID- 9211107 TI - [Severe aortic regurgitation with marked thickening of the aortic annulus: a case report]. AB - A 74-year-old woman with severe aortic regurgitation and marked thickening of the aortic annulus received aortic valve replacement. Transesophageal echocardiography demonstrated marked thickening of the aortic annulus. The maximum thickness of the aortic annulus was about 11 mm at the side of the right coronary cusp. Histological examination revealed non-specific inflammation of the aortic annulus and aortic valve chiefly composed of lymphocytes and plasma cells. Thickening of the aortic annulus associated with inflammatory aortitis was the cause of aortic regurgitation in this patient. PMID- 9211108 TI - [Successful surgical treatment of aortic regurgitation due to annuloaortic ectasia and mitral regurgitation caused by tendon rupture in a case of osteogenesis imperfecta]. AB - A 54-year-old man presented with osteogenesis imperfecta complicated with both aortic regurgitation due to annuloaortic ectasia and mitral regurgitation secondary to tendon rupture. He had spinal and carpal deformities in his childhood, and heart murmurs were identified at the age of 25. He was admitted complaining of dyspnea on effort. His height was 142 cm and his weight was 46 kg. He had kyphosis, scoliosis and carpal deformity. Blue sclera was not observed. Chest radiography showed cardiomegaly and lung congestion. Echocardiography showed annuloaortic ectasia, mild aortic regurgitation, and serious mitral regurgitation due to postero-apical tendon rupture. Bone deformity and his statues were indicative of osteogenesis imperfecta. He received modified Bentall and mitral valve replacements. PMID- 9211109 TI - [Preventive concomitant aortic root replacement for annuloaortic ectasia in a patient with Marfan syndrome undergoing mitral valve replacement for mitral regurgitation]. AB - A 28-year-old woman presented with Marfan syndrome combined with severe mitral regurgitation and annuloaortic ectasia. The ascending aorta was dilated to 48 mm in diameter without aortic regurgitation. Considering the increased operative risk due to complication with aortic dissection, simultaneous replacement of the mitral valve and aortic root were performed. Her postoperative course was uneventful. Several options of the surgical treatment for Marfan syndrome are discussed. PMID- 9211110 TI - [Intraoperative and follow-up color Doppler echocardiography for evaluating the surgical results of aortic valvuloplasty in a patient with bicuspid aortic valve]. AB - A 21-year-old man underwent aortic valvuloplasty for aortic regurgitation due to bicuspid aortic valve. After the initial procedure for valvuloplasty of the anterior cusp, transesophageal color Doppler flow imaging revealed a significant regurgitant jet directed posteriorly as observed preoperatively. After an additional valvuloplastic procedure for the anterior cusp, the size of the regurgitant signal was reduced and directed to the center of left ventricular outflow tract. The surgery was then completed. On the 5th postoperative day, an aortic regurgitant murmur reappeared and color Doppler echocardiography showed a mild but significant regurgitant jet directed toward the interventricular septum, which may have been due to contraction of the anterior cusp treated by valvuloplasty. Aggravation of the aortic regurgitation in the early stage after plastic surgery is important and requires consideration in the intraoperative assessment. PMID- 9211111 TI - [Concealed infective endocarditis]. AB - The occurrence of concealed infection or "natural remission" in infective endocarditis was investigated microscopically in 35 consecutive patients (36 valves) with infective endocarditis (between April 1987 and May 1995). Four patients were considered as having concealed or silent infective endocarditis. Preoperative diagnosis of these patients was mitral valve prolapse, rheumatic aortic valve stenosis with insufficiency, aortic valve prolapse and hypertrophic cardiomyopathy, respectively. These patients did not present with any clinical signs of infective endocarditis such as cardiac murmur and ventricular dysfunction. Histological examination of the excised valves revealed valvular perforation, small round cell infiltration, neovascularization, remnants of vascular smooth muscle cells, and organizing vegetations. These findings are consistent with the histological findings of infective endocarditis. Latent infective endocarditis may be present without inflammatory manifestation. PMID- 9211112 TI - [Echocardiographic prediction of risk for embolism in patients with infective endocarditis]. AB - The relationship between two-dimensional echocardiographic findings of vegetation and embolic events was investigated in 26 patients with infective endocarditis (17 males and 9 females, mean [+/-SD] age 51 +/- 17 years). The size and the other morphologic characteristics of vegetation (mobility, extent and consistency) were analyzed retrospectively according to the criteria by Sanfilippo, et al., and parameters were assigned scores from 1 to 4 to provide a total score. Patients with a maximum vegetation diameter > 10 min had a significantly higher incidence of embolic events than those with < or = 10 mm (p < 0.05). Each parameter of vegetation showed no significant difference between patients with and without embolic events; but the total score was significantly higher in patients with embolic events (p < 0.05). Particularly, all patients with a total score > or = 10 had embolic events, whereas those without embolic events had a total score < or = 9. There were no significant differences in the frequency of emergent valve replacement between patients with aortic value and mitral valve endocarditis. However, the incidence of heart failure was higher, but not significantly, in patients with aortic valve (67%) and combined valve endocarditis (67%) than in those with mitral valve endocarditis (36%). The maximum size and total score reflecting mobility, extent and consistency of vegetation using two-dimensional echocardiography provide useful information to predict the occurrence of embolic events in patients with infective endocarditis. PMID- 9211113 TI - [A patient with mitral stenosis due to infective endocarditis]. AB - A 51-year-old woman presented with mild stenosis of the mitral valve which had become thickened and rigid due to infective endocarditis, manifesting as persistent fever of up to 40 degrees C and general fatigue of a few days' duration. A harsh systolic murmur was heard. Multiple blood cultures revealed alpha-streptococcus. Echocardiography disclosed asymmetric septal hypertrophy (interventricular septal thickness/posterior wall thickness, 19/14 mm) and systolic anterior wall motion of the mitral valve. Continuous wave Doppler ultrasonography showed a peak left ventricular outflow tract pressure gradient of 170 mmHg. Transesophageal echocardiography revealed vegetations on the anterior mitral leaflet, aortic valve and interventricular septum along the left ventricular outflow tract. In particular, the anterior mitral leaflet was thickened and moved poorly. The calculated mitral valve areas was 1.5 cm2 and peak diastolic left atrium-left ventricle pressure gradient was 7 mmHg. A specimen of the mitral valve did not reveal commissural adhesion, but the anterior mitral leaflet showed marked fibrous thickening caused by scarred vegetation. Based on these findings, the diagnosis was hypertrophic obstructive cardiomyopathy complicated by infective endocarditis and "mitral stenosis". Valvular regurgitation is a common complication of active and healed infective endocarditis. In contrast, infective endocarditis rarely causes valvular stenosis except for stenosis caused by large fungus vegetation. PMID- 9211114 TI - [An elderly patient with infectious endocarditis complicated with congestive heart failure due to mitral and tricuspid regurgitation]. AB - A 77-year-old man was referred to our hospital on October 2, 1995 because of fever and left mandibular pain beginning three months before admission. His blood pressure was 90/60 mmHg. A grade III/VI pansystolic murmur was heard over the cardiac apex. The liver was palpable 4 cm below the right costal margin. Lower extremity edema was present bilaterally. White blood cell count was 7,030/mm3 and C-reactive protein was 2.54. Enterococcus faecalis was identified by the blood culture. The diagnosis was infective endocarditis associated with congestive heart failure. He was treated by administration of antibiotics and diuretics. Mitral valve replacement and tricuspid annuloplasty were performed on October 19 because of progressive congestive heart failure with oliguria. The surgical intervention was successful despite the presence of multiple risk factors: high age, emergency, congestive heart failure and active infection. His condition improved dramatically after the operation and he was discharged two months later. Surgical intervention for infective endocarditis was a significant high-risk procedure in this uncontrollable and elderly case. This successful result suggests the indication for the timing of surgery. PMID- 9211115 TI - [Mitral prosthetic valve replaced twice due to repeated prosthetic valve endocarditis: a case report]. AB - A 38-year-old man was admitted to our hospital for detailed examination of fever, cough and yellow sputum. At the age of 32, be had mitral prosthesis for the first time, because of mitral regurgitation due to mitral valve prolapse. Four years previously, he had again undergone mitral prosthetic valve replacement due to prosthetic valve endocarditis due to staphylococcus epidemidis. This occasion, staphylococcus aureus was isolated by arterial blood culture. Transesophageal echocardiography detected vegetation attached to the mitral prosthetic valve and paravalvular leakage. The diagnosis was prosthetic valve endocarditis. He underwent a third mitral prosthetic valve replacement. Detection of the source of infection was difficult only by transthoracic echocardiography, and immediate transesophageal echocardiography seemed mandatory to diagnose bacterial endocarditis. PMID- 9211116 TI - [Aortic valve insufficiency caused by nonpenetrating chest trauma difficult to distinguish from infective endocarditis with transesophageal echocardiography: a case report]. AB - A 58-year-old man was involved in an automobile accident and suffered remittent fever, leukocytosis and high C-reactive protein level. He developed a diastolic murmur 2 months after the accident. Transesophageal echocardiography showed severe aortic regurgitation with a vegetation-like echo image attached to the right coronary cusp leaflet, suggesting infective endocarditis. Intensive medical treatment for 11 months did not improve the vegetation-like echo-image, so aortic valve replacement was performed. Disruption of the right coronary cusp leaflet was confirmed surgically. Prolapse had occurred as a result of disruption during diastole. The vegetation-like echo-image was considered to be the tip of this leaflet. PMID- 9211117 TI - Retinoic acid exposure of the mouse on embryonic day 9 selectively spares derivatives of the frontonasal neural crest. AB - Retinoic acid is known to perturb craniofacial development and can be used to understand processes controlling early embryonic development of the face. The effects of retinoic acid on mouse craniofacial development were studied by administration of a single dose (25-200 mg/ kg) of all-trans retinoic acid (RA) to timed pregnant C57BL6/J mice at gestational days (gd) 8.25, 9, or 10. RA exposure on gd 8.25 or gd 10 resulted in craniofacial defects in fetuses but gd 9 exposure revealed a differential effect of RA depending upon whether tissues were derived from branchial arch or frontonasal neural crest. Embryos exposed to RA at gd 9 showed a dose-dependent effect of RA on branchial arch derived tissues; first arch derivatives were most severely affected with the mandible and zygoma becoming severely dysplastic at the highest dose of RA (200 mg/kg). However, RA exposure on gd 9 completely spared frontonasal neural crest-derived tissues. Paired premaxillae nasal and frontal bones as well as the cartilaginous nasoethmoid region and nasal capsule containing the osseous vomer showed no statistical difference from those of control animals. These studies showed a temporal and differential sensitivity to RA and may suggest a developmental heterogeneity of the cephalic neural crest cells destined to participate in formation of craniofacial structures. PMID- 9211118 TI - 3D-computed tomography: a new method for the evaluation of fetal cranial morphology. AB - This study is the first presentation of three-dimensional computed tomography (3D CT) for the in vitro evaluation of the prenatal human cranium. The study was based on CT examinations from 26 aborted normal fetuses between 10 and 25 weeks gestational age. Incremental coronal and transverse CT slices of 1 mm thickness and a threshold segmentation algorithm were used to generate 3D-CT reconstructions (surface-shaded display, SSD) of the cranial bones similar to their anatomical appearance. The threshold of the segmentation algorithm was selected after comparison of the 3D-CT images generated with varying thresholds and graphically reconstructed histological serial sections of particular sutures in five specimens. The variation of the segmentation threshold resulted in alterations of the bone sizes and suture widths. However, 3D-CT images allowed sensitive identification of the cranial ossification centers and accurate evaluation of the bone topography. Cutting and rotating procedures made it possible to evaluate all imaged bones in arbitrary views without disturbing superpositions, thus making isolated examinations of particular macroscopic sections of the specimens unnecessary. In conclusion, 3D-CT of the fetal cranium promises to be of considerable help in the evaluation of prenatal cranial development. PMID- 9211119 TI - A new craniofacial disorder involving hypertelorism and malformations of external nose, palate and pituitary gland. AB - The aim of the present study was to describe and pathologically evaluate an apparently unreported craniofacial malformation, based on comparison of the cranial midsagittal components with similar components under normal developmental conditions. A severely malformed fetus with a gestational age of about 17 weeks underwent whole body and special craniofacial radiography. Following autopsy dissection, the midsagittal segment of the cranial base, including the eyes, was radiographed in different projections. Midsagittal tissue blocks were serially sectioned for microscopy. Routine stains and immunohistochemical stains were applied. The face was characterized by hypertelorism, absence of external nose but with open shell-like cavities medio-cranially to the eyes, and by a palate fused in the midline and with extensive bony ridges laterally. There was absence of normal nasal cavities, presence of nasal septum and vomer, normal eyes, and nasal ducts covered with nasal mucosa ending blindly in the cartilage. No olfactory bulbs were found. The palatal ridges consisted of bony tissue. The pituitary gland was severely malformed and consisted solely of adenopituitary gland tissue, located in its full extent in the pharyngeal mucosa. There was no sella turcica. From a pathogenetic point of view, it is suggested that the neural crest cells in the frontonasal region of the crest were reduced in amount or late in migration to the midfacial region compared to the neural crest cells to the maxillary region. Therefore, we believe that the malformations observed in the nasal placodes and in the pituitary placode, combined with abnormal migration or abnormal timing of neural crest cells during the craniofacial development, are important factors behind this disorder. PMID- 9211121 TI - The feet in Crouzon syndrome. AB - Eighteen patients with Crouzon syndrome were evaluated for anomalies of the feet. Clinical examination was unremarkable in all cases. Radiographs were evaluated by a radiologist with an interest in skeletal dysplasia, along with the craniofacial team. A range of radiographic anomalies was seen, with the phalanges, metacarpals, and tarsals all displaying anomalies. Only three cases had radiographically normal feet. These findings suggest that the effects on the feet, which, although subtle and not well described in the literature, are notable. Feet anomalies also occur with the other complex craniosynostosis syndromes resulting from mutations of fibroblastic growth factor receptor 2 molecule, such as those of Apert, Pfeiffer, and Jackson-Weiss syndromes. PMID- 9211120 TI - Oculo-auriculo-vertebral spectrum: cranial and vertebral malformations due to focal disturbed chondrogenesis. AB - Microradiographic and histological analyses point out a focal disturbed chondrogenesis of both the skull base and the axial skeleton in a case of oculo auriculo-vertebral spectrum. Cartilage showed disturbed endochondral ossification with defects in calcification, deficient resorption, and abnormal crumpled areas of mineralized cartilage. PMID- 9211123 TI - Subclinical Graves' disease as a cause of subnormal TSH levels in euthyroid subjects. AB - In order to elucidate causes of subclinical thyrotoxicosis, we reviewed records of thyroid function tests obtained in our hospital between 1990 and 1992 showing normal thyroid hormones and subnormal TSH levels, and analyzed underlying clinical conditions of the patients. Of 186 patients with normal T4 and/or free T4 and normal T3 and/or free T3 but subnormal TSH (< 0.1 mU/l) levels in serum, 150 were under treatment with antithyroid drugs for hyperthyroid Graves' disease or with thyroid hormones for hypothyroidism. Twelve were in remission after treatment for Graves' disease, and 4 had destructive thyroiditis. Of the remaining 20 patients, 4 had autonomously functioning thyroid nodule (AFTN), 9 had euthyroid ophthalmic Graves' disease (EOG), and 7 had diffuse goiter without apparent ophthalmopathy (DG). When thyroid stimulating antibodies (TSAb) were measured in the last 3 groups of the patients, they were detected in none with AFTN but in all patients with EOG and DG. These 7 DG patients without ophthalmopathy had a clinical feature showing unstable thyroid functions, changeable to euthyroidism, overt hyperthyroidism and even hypothyroidism during follow-up. In conclusion, TSAb measurement is useful for detection of subclinical Graves' disease in euthyroid subjects with subnormal TSH levels in serum. PMID- 9211122 TI - Bilateral adrenal enucleation-induced changes in adenohypophyseal pro opiomelanocortin (POMC)-related peptides synthesis and secretion: a comparative study with adrenalectomized rats. AB - The aim of the present study was to elucidate the modulatory effect of transient changes in endogenous glucocorticoids, occurring after bilateral adrenal enucleation (ENUC), on anterior pituitary (AP) proopiomelanocortin (POMC)-derived peptides synthesis and output in rats. For this purpose, adult female rats were either bilaterally ENUC, adrenalectomized (ADX), or sham-operated (SHAM) and killed by decapitation 2, 7, 14, and 21 days after surgery. Trunk blood was collected for measurements of ACTH, beta-endorphin (beta-END) and corticosterone (B) concentrations; APs were quickly dissected for the determination of ACTH, beta-endorphin (beta-END)-like (beta-END-LI) and gamma 3-MSH contents and adrenal glands were removed and submitted to histological study. The results indicate that ENUC and ADX increased AP POMC-related peptides synthesis and release in association with changes in the AP processing of peptides belonging to the N terminal (gamma 3-MSH), mid (ACTH) and C-terminal (beta-LPH/ENDs) portions of POMC. While ADX abolished plasma B levels, ENUC induced a transient (day 2) decrease in plasma B concentrations which returned to SHAM levels at 7 days after surgery. These data tallied with the histological observations carried out, indicating a time-dependent regenerative process of the adrenal which was completed by three weeks after ENUC. There was a different pattern in plasma ACTH and beta-END levels between ENUC and ADX; maximal plasma peptide levels were found 7-14 days after ENUC, then falling down to SHAM values at 21 days post ENUC. Conversely, there was a constant increment in plasma peptide levels up to 21 days after ADX. At 2 days after both ENUC and ADX all peptides measured in the AP were lower than SHAM values, thus reflecting a rapid corticotrope secretion. Thereafter, 7 or more days after surgery, AP peptide content in ADX rats increased, in a time-related fashion, up to 21 days after surgery. Only beta-END LI showed a similar AP content to that of the SHAM group, thereafter indicating a preferential cleavage of POMC to beta-END long after ADX (21 days). ENUC rats showed increased AP POMC peptides content throughout the whole time, and it was significantly different from SHAM and ADX values 14 days post-surgery. Interestingly, we found an increment in AP gamma 3-MSH, a peptide which is preferentially synthesized in the intermediate lobe of the rat pituitary, in both ENUC and ADX situations. Our results further indicate that: 1) glucocorticoids, from regenerating adrenal origin, induce a fast negative feedback mechanism on AP secretion, and 2) there might be a delayed inhibitory action of newly synthesized corticosteroids on higher levels of the central nervous system. The lack of glucocorticoids (ADX) clearly corroborates a persistent enhancement of AP POMC related peptides synthesis and secretion. The differences in AP processing of POMC between ENUC and ADX might be due to qualitative/quantitative changes in hypothalamic ACTH secretagogues output. PMID- 9211124 TI - CYP17 gene analysis in hyperandrogenised women with and without exaggerated 17 hydroxyprogesterone response to ovarian stimulation. AB - Nine patients with polycystic ovary syndrome (PCOS) and exaggerated serum level of 17-hydroxyprogesterone (17-OHP) response to gonadotropin (GnRH) agonist stimulation, and seven patients with PCOS, but normal 17-OHP response have been analysed for possible linkage of PCOS with genetic defects on the 17 alpha hydroxylase/17,20-lyase gene (CYP17). A portion of the regulatory and the entire coding domain for this enzyme have been analysed by PCR-SSCP analysis. Samples have been also screened for the previously reported -34 bp polymorphism, which creates a new SP1-type promoter site, but was excluded as the primary genetic defect. We have varied gel concentrations, reduced running temperatures, added glycerol to polyacrylamide gels and performed electrophoresis on longer gels in order to improve the resolving power of SSCP. Screening of the CYP17 gene revealed no mutations associated with the disease in the examined group of patients. Also, the -34 bp polymorphism proved to be equally distributed among patient and control samples, which in our case were non-related. The results indicate that, when germline mutations in question, CYP17 may be excluded as a candidate gene for these subtypes of PCOS. PMID- 9211125 TI - LH isoform profiles during short-term pulsatile LHRH administration in elderly men. AB - LH isoform profiles were analyzed in sera resolved with isoelectrofocusing from 5 elderly men (age 70.6 +/- 2.95) and 5 young adult men (age 28.2 +/- 1.24), by using polyclonal antibodies (RIA), monoclonal antibodies directed against the beta-subunits (IRMA) and in vitro LH bioassay. Despite the fact that the elderly had lower testosterone levels than the young (293 +/- 38 vs 512 +/- 77 ng/dl, p < 0.05), no differences were noted in the isoforms detected by any of the assays, although each assay yielded a characteristic profile. Indeed, RIA showed most LH in the acidic range, while IRMA revealed LH profiles with a major peak in the basic range, thus resembling the profiles determined by means of the bioassay. In the elderly, the profiles were also analyzed on day 7 and day 14 of short-term pulsatile sc LHRH administration (150 ng/bw/120 min). Only the LH bioassay detected an LHRH-induced shift to more basic and bioactive forms; these changes accompanied an increased in testosterone levels on day 7 (396 +/- 83 ng/dl, p < 0.05 vs day 0) and on day 14 (320 +/- 58 ng/dl NS vs day 0). Our data suggest that: 1) the profiles obtained in young and elderly subjects are similar, irrespective of the antisera used; 2) as a result of treatment with LHRH in the elderly an increase in T levels occurs, possibly due to the observed changes in LH bioactivity; 3) the in vitro LH bioassay appears to be the most sensitive assay in detecting such changes, which consisted of an enrichment in more basic and bioactive glycoforms. PMID- 9211126 TI - Effects of estradiol benzoate and progesterone on superoxide dismutase activity in the rat liver. AB - Activities of superoxide dismutases MnSOD and CuZnSOD were measured in appropriate subcellular fractions prepared from livers of intact and long-term gonadectomized (GX) rats of both sexes, and of GX female and male rats injected sc with a single dose of 5 micrograms estradiol benzoate (EB) or 2 mg progesterone (P). In female livers, MnSOD activity did not vary significantly during the estrous cycle, declined after gonadectomy in comparison to proestrus, and was steady in GX females treated with EB or P. The activity of CuZnSOD was lowered at proestrus and elevated after removal of the ovaries in comparison to proestrus value. EB suppressed, and P elevated CuZnSOD activity in GX females. In the liver of male rats, MnSOD was not affected by gonadectomy nor by EB and P treatments. CuZnSOD activity was reduced following orchiectomy and enhanced in GX males following treatment with P, while EB had no effect. These results suggest that P and EB modulate the activity of CuZnSOD and do not affect MnSOD in the rat liver. The modulatory effects are elicited by P in the males and by P and EB in the females. PMID- 9211127 TI - Evidence for an interaction between alpha-MSH and opioids in the regulation of gonadotropin secretion in man. AB - Gonadotropin secretion is inhibited by the endogenous opioids and stimulated by their antagonist naloxone. LH secretion is stimulated by alpha-MSH, a tridecapeptide derived from the post-translational processing of POMC. The possibility that alpha-MSH interacts with the opioids, as suggested by the experimental evidence, was investigated in 7 normal males aged 24-29 through the performance of seven tests: naloxone (0.8 mg i.v. bolus, followed by infusion of 1.6 mg/h for 120'); alpha-MSH (2.5 mg i.v. bolus); naloxone + alpha-MSH (2.5 mg i.v. 15' after commencement of the naloxone infusion); naloxone + GnRH (100 micrograms i.v. 15' after commencement of the naloxone infusion); alpha-MSH + GnRH (respectively 2.5 mg and 100 micrograms at time 0), GnRH alone (100 micrograms at time 0), placebo (150 nmol/l NaCl solution). The LH AUCs during both naloxone (30.3 +/- 2.7 mIU/ml.min-1) and alpha-MSH test (32.9 +/- 4.6 mIU/ml.min-1) were significantly greater (p < 0.005) than that observed during placebo (16.9 +/- 3.6 mIU/ml.min-1). The LH AUC during alpha-MSH + naloxone (37.6 +/- 2.6 mIU/ml.min-1) was not significantly different from that recorded during their separate administration. GnRH injected alone, during the naloxone infusion and with alpha-MSH produced similar increases in LH, that were significantly higher than that observed during the other tests (AUCs: GnRH 89.4 +/- 10.6, GnRH + naloxone 100.5 +/- 9.1, GnRH + alpha-MSH 94.6 +/- 7.9 mIU/ml.min-1, p < 0.001). Significant increase in FSH (p < 0.001) was only observed during GnRH, GnRH + naloxone and GnRH + aMSH tests (AUCs: placebo 13.3 +/- 1.7; naloxone 14.7 +/- 2.5; alpha-MSH 15.5 +/- 2.3; alpha-MSH + naloxone 16.9 +/- 1.9; GnRH 19.1 +/- 1.1; GnRH + alpha-MSH 20.7 +/- 1.3; GnRH + naloxone 21.2 +/- 1.8 mIU/ml.min-1). These results are in line with the possibility of an interaction between alpha MSH and the opioids in the regulation of gonadotropin secretion, perhaps with opposing effects on a final common pathway. PMID- 9211128 TI - Recurrent thromboembolism as a hallmark of Cushing's syndrome. AB - The present report describes a 54-year-old woman with a history of recurrent thromboembolic events. The clinical and physical examination led to suspect Cushing's syndrome. Screening tests (urinary free cortisol excretion and 1 mg dexamethasone) were inconclusive, but a detailed endocrine work up confirmed the presence of ACTH-dependent hypercortisolism. The patient was cured by the removal of a ACTH-secreting microadenoma by transsphenoidal route. The present case provides a clinical demonstration of a previous experimental evidence that a hypercoagulable state is present in Cushing's syndrome. PMID- 9211129 TI - Thyroid function in children treated for acute lymphoblastic leukemia. AB - To determine the late effects of treatment on thyroid function in children who survive acute lymphoblastic leukemia, we assessed plasma levels of thyroid hormones in 24 children (15 girls and 9 boys) who had received combination of chemotherapy along with 18-24 Gy of irradiation to the cranium. The children were aged between 1 and 10.5 years (mean 3.1) at diagnosis and thyroid status was investigated between 1.3 and 13 years (mean 6.8) after completion of therapy. Six children showed a low peak of plasma thyroid stimulating hormone (TSH), after stimulation with thyrotrophin-releasing hormone (TRH). Three children showed a low basal plasma TSH concentration. Serum levels of thyroxine (T4, fT4) and triiodothyronine (T3, fT3) were normal in all patients. The frequency of thyroid hypofunction (low peak response of TSH to TRH) was more common in children receiving 24 compared to 18 Gy cranial irradiation (50% vs 14%; odds ratio = 7) and those who had completed therapy more than 5 years ago (31.3% vs 12.5%, odds ratio 3.18) although no significant association could be found (95% IC: 0.27-65.8 and 0.24-90 respectively). Because of the low mean age at diagnosis of our population no significant association could be found between children younger than 3 years of age at diagnosis and thyroid hypofunction (odds ratio = 0.14; 95% IC: 0.01-1.48). We conclude that treatment for acute lymphoblastic leukemia including cranial irradiation may lead to TRH/TSH dysfunction and therefore long term survivors should be followed for a long period after completion of therapy. PMID- 9211130 TI - Effects of ovary suppression by a long-acting GnRH-agonist on circulating GH, insulin-like growth factor I and insulin levels in women with polycystic ovary syndrome. AB - Our aim was to investigate the effect of GnRH-agonist (GnRH-a) induced suppression of plasma sex steroids on serum GH, insulin like growth factor-I (IGF I) and insulin levels after an oral glucose load (OGTT) in women with polycystic ovary syndrome (PCOS). Serum insulin, GH and IGF-I levels during a 75-g 4-h OGTT were measured in 3 nonobese and 7 obese hyperandrogenic women with PCOS and normal glucose tolerance before and after 10 weeks of treatment with the GnRH-a triptorelin (3,75 mg im every 28 days). Basal estrogen and androgen levels were also measured at time 0 of the first and the second OGTT. After the therapy serum estrogens and androgens were significantly suppressed. Body weight remained unchanged. Basal GH significantly increased after the treatment while fasting IGF I and insulin levels decreased from (mean +/- SE) 349.3 +/- 31.8 to 278.7 +/- 33.2 ng/mL and from 22.4 +/- 4.1 to 18.8 +/- 4.4 microU/mL, respectively. The insulin response to OGTT (area under curve) was also reduced (from 16,017 +/- 2598 to 11,736 +/- 2317 microU/mL/240 min). Our results suggest that the GnRH-a induced suppression of ovary secretion may modify the serum GH and IGF-I levels and the insulin response to an OGTT in women with PCOS. PMID- 9211131 TI - In vivo antiestrogenic activity of mifepristone in the rat. AB - The aim of this study was to find out whether mifepristone, known mainly as a substance with an antiprogesterone and antiglucocorticoid effect, also has an in vivo antiestrogenic activity on the adenohypophysis of the rat. Male Wistar rats were given chronically either estradiol-benzoate (EB, 1 mg s.c. twice a week) for a period of 12 days, or the non steroidal antiestrogen tamoxifen (1 mg/day/rat), or mifepristone (1 mg/day/rat), or EB together with mifepristone or tamoxifen. The hypertrophic effect of the EB on the weight of the adenohypophysis (AP) was significantly suppressed both by tamoxifen and by mifepristone. Mifepristone and tamoxifen reduced the increased content of PRL in the estrogenized adenohypophysis. Mifepristone but not tamoxifen significantly increased the content of the LH in the adenohypophysis of estrogen treated rats. Mifepristone and tamoxifen suppressed the increased concentration of cyclic nucleotides cAMP and cGMP in the estrogenized adenohypophysis. Mifepristone given alone increased serum levels of corticosterone, but when given together with EB deepened inhibiting effect EB on them. The results of our preliminary study show that mifepristone exerts a weak antiestrogenic activity on the level of hypophysis, however the pharmacology is not identical to tamoxifen. PMID- 9211133 TI - Hyperosmolar nonketotic coma at the onset of type I diabetes in a child. AB - A 12 yr-old child without any past medical history of diseases was admitted to hospital for sopor and polyuria. At admission he was markedly dehydrated. Blood glucose was 72 mmol/l, sodium 154 mmol/l, osmolarity 381 mOsm/Kg, urinary ketons were negative. He was rehydrated with hypotonic saline and treated with insulin. The osmolality and sodium initially increased to 176 mmol/l and 408 mOsm/Kg respectively and progressively decreased to normal levels. Serum transaminases increased to GOT 336 and GPT 209 U/l in the first days of treatment and normalized after 15 days. The anti-islet antibodies were positive. The non ketotic hyperosmolar coma and Type I diabetes is rare in children but this possibility must be kept in mind especially when some familial or psychological problems are present as in our case. PMID- 9211132 TI - Adrenocorticotropin-producing pituitary carcinoma with liver metastasis. AB - We report here the extremely rare case of a twenty-eight year-old woman with a metastatic ACTH-secreting pituitary carcinoma. This is the thirteenth case to be described in the literature. Ten years ago Cushing's disease was diagnosed. After pituitary surgery, then bilateral adrenalectomy, a Nelson's syndrome appeared. The particularly extensive pituitary secondary development led to several pituitary surgical procedures, radiotherapy, and octreotide treatment. Eight years after Cushing's disease was diagnosed, liver tumors were discovered. Pathological examination and ACTH immunostaining demonstrated the secretory nature of these metastases. The lack of ectopic tumor, the LPH/ACTH equimolar ratio and a study of the plasma proopiomelanocortin derivatives by HPLC showed that the ACTH secretion originated in pituitary tissues (in situ and liver metastases). The processing of POMC seems thus to be normal in this kind of tumor and metastases. Intact POMC levels were very high, indicating an aggressive tumor, and ACTH/LPH production was paradoxically stimulated by octreotide. This case is also exceptional because of the slow development of the disease, which may be due to the complementary hepatic chemoembolization treatment. PMID- 9211135 TI - Beta-blockers and ocular blood flow: a perspective. PMID- 9211134 TI - Two-step development of a pituitary adenoma: from hyperprolactinemic syndrome to Cushing's disease. AB - In this report we describe the case of a young female patient with amenorrhea galactorrhea syndrome apparently due to pituitary PRL-secreting adenoma who, after three years of dopaminergic therapy without any shrinkage of the tumor, developed true Cushing's disease. Progression from hyperprolactinemia to hypersecretion of ACTH has been rarely described and it may be due to different possibilities. However, histopathological and immunohistochemical studies of the adenoma showed a pattern of PRL negative and ACTH positive cells, excluding mixed pituitary tumor. In order to explain the progression from hyperprolactinemia with amenorrhea-galactorrhea to an ACTH hypersecretion syndrome, it must be hypothesized either pituitary stalk compression or the influence of paracrine regulation factor(s) (such as Galanine) due to an "initially silent" corticotropinoma. This case confirms that the presence of hyperprolactinemia in a patient with pituitary tumor and amenorrhea-galactorrhea syndrome is insufficient to confidently conclude for prolactinoma. Furthermore, it underlines the importance both of clinically monitoring the patient with prolactin pituitary adenoma if dopaminergic therapy does not reduce tumor volume, and of accurately and repeatedly measuring the other pituitary hormonal secretions. PMID- 9211136 TI - The effect of a brief education program on glaucoma patients. AB - PURPOSE: To assess our patients' knowledge of glaucoma and to measure the effect of a brief education program on their understanding of glaucoma. METHODS: Patients attending glaucoma clinics at a university and a Veterans' Affairs hospital were randomized into two groups: "exposed" and "unexposed" to a simple education program of a video and brochures. Glaucoma knowledge was assessed twice by an oral questionnaire, at 2 weeks and 6 months after randomization plus or minus education. RESULTS: Younger patients and those with more years of formal schooling knew more about glaucoma. Two weeks after the education program, the exposed group performed significantly better than did the unexposed group. Analysis of the results showed benefit from both brochures and video. This effect of education was not seen at retesting 6 months later. CONCLUSION: Older patients and those with less formal education know less about glaucoma. A brief, simple education program can significantly improve levels of knowledge about glaucoma, even in a relatively well-informed population. However, patient education must be repeated to maintain a useful effect. PMID- 9211137 TI - Primary valve malfunction of the Krupin eye valve with disk. AB - PURPOSE: To examine the management and possible causes of primary valve malfunction of the Krupin eye valve with disk. METHODS: The authors reviewed the results of 113 patients undergoing implantation of the Krupin eye valve with disk and identified eight patients with primary valve malfunction requiring surgical revision. RESULTS: Valve revision involved manipulation (n = 1 case), explantation of the malfunctioning valve and implantation of a new valve (n = 2), and amputation of the valve (n = 5). Six of eight patients had final intraocular pressures of < 21 mmHg on one or no medications at a mean interval of 15.9 months (range 5-36) after surgical revision. Transient postoperative hypotony was noted in three patients and chronic hypotony with loss of light perception in one patient. One explanted valve was examined and found to have partially fused leaflets. CONCLUSIONS: Surgical revision in cases of primary valve malfunction of the Krupin eye valve with disk may be accomplished relatively safely with an acceptable level of postoperative complications. The etiology of primary valve malfunction may be related to the sterilization process and prolonged storage before implantation. PMID- 9211138 TI - Reproducibility of retinal and optic nerve head blood flow measurements with scanning laser Doppler flowmetry. AB - PURPOSE: To evaluate the interobserver variability and the reproducibility of retinal and optic nerve head capillary blood flow measurements performed with a new noninvasive equipment, the scanning laser Doppler flowmeter (Heidelberg Engineering, Heidelberg, Germany). METHODS: Blood flow measurements were performed during three independent sessions in six patients with glaucoma and five normal subjects using the scanning laser Doppler flowmeter (SLDF), which allows the visualization of perfused capillaries and vessels of the retina and optic nerve head and enables the quantification of capillary blood volume, flow, and velocity in any selected area of the perfusion map. To evaluate the interobserver variability in selecting the areas in the perfusion map to be measured, three observers tried to locate the same areas in the perfusion map of images obtained during the first session. To evaluate the reproducibility of the measurements, the observers measured correspondent areas in the peripapillary retina and in the optic nerve head of images from the three sessions. Areas of different sizes (10 x 10 pixels and 4 x 4 pixels) were measured. RESULTS: The agreement between readings performed by the three observers was very good, with the reliability coefficient for the various parameters varying from 0.90 to 0.98. The reproducibility of retinal and lamina cribrosa measurements with the 10 x 10 pixel square target was good (reliability coefficient for the different parameters ranging from 0.70 to 0.85) and much better than the reproducibility of the 4 x 4 pixel target. The measurements performed in the neuroretinal rim area also had poor reproducibility. The measurements from the patients with glaucoma tended to be more reproducible than those from normal subjects. CONCLUSIONS: The SLDF allows reproducible blood perfusion measurements of retinal and lamina cribrosa areas when a target square of 10 x 10 pixels is used. PMID- 9211139 TI - Appositional angle closure in eyes with narrow angles: an ultrasound biomicroscopic study. AB - PURPOSE: To demonstrate appositional angle closure using ultrasound biomicroscopy under dark room conditions, to investigate patterns of appositional closure, and to correlate those patterns with the angle topology in eyes with narrow angles. METHODS: Forty-six eyes that met the study criteria, from 46 individuals, were examined by ultrasound biomicroscopy under dark room and illuminated conditions. RESULTS: In 40 of the 46 eyes (87.0%), appositional closure was identified by ultrasound biomicroscopy. Two distinct patterns of angle closure were seen: type B, in which closure starts from the bottom of the angle, and type S, in which closure starts in the vicinity of Schwalbe's line. In type B, the iris root was located closure to the chamber angle than in type S. CONCLUSIONS: Ultrasound biomicroscopy performed under dark room conditions is a useful technique for identifying appositional angle closure in eyes with narrow angles. The topology of the iris root is related to the pattern of the appositional angle closure. PMID- 9211140 TI - Nocturnal ophthalmic arterial hemodynamics in primary open-angle glaucoma. AB - PURPOSE: Recent studies have found nocturnal reductions in systemic arterial blood pressure associated with progressive visual field loss in glaucoma. Although ocular ischemia has been hypothesized to link these two phenomena, it remains unknown if perfusion of the eye is reduced during the night in patients with glaucoma. PATIENTS AND METHODS: Nine patients with primary open-angle glaucoma (POAG) and stable visual fields who were free from systemic hypertension, as well as nine age- and gender-matched controls, were studied at 9:00 P.M., and then during sleep at 12:00, 3:00, and 6:00 A.M. Systemic blood pressure, intraocular pressure (IOP), and color Doppler imaging (CDI) of the ophthalmic artery were measured at each time. RESULTS: Arterial blood pressure and ophthalmic artery peak systolic and end-diastolic velocities were similar and were unchanged over time, in both groups. In contrast, the ophthalmic arterial resistance index decreased as the night progressed (p < 0.05), identically in controls and patients. In patients with glaucoma, CDI indices were independent of changes in arterial pressure, IOP, or calculated ocular perfusion pressure. CONCLUSION: Patients with POAG characterized by stable visual fields who were free from systemic hypertension exhibited normal ophthalmic arterial hemodynamics at night; there was no evidence of ocular ischemia or vasoconstriction. PMID- 9211141 TI - Pulsatile ocular blood flow measurements in healthy eyes: reproducibility and reference values. AB - PURPOSE: To determine the reproducibility and the normal reference range of pulsatile ocular blood flow (POBF) values in healthy subjects using the Ocular Blood Flow Tonograph (OBF Laboratories, UK Ltd., Wiltshire, England). METHOD: Pulsatile ocular blood flow was measured in one eye of each of 83 patients. Coefficient of reliability was determined by calculation of intraclass correlation coefficient via one-way analysis of variance. Mean difference between measurements was calculated for bias and first exposure effects. Pulsatile ocular blood flow from 163 healthy individuals were analyzed to determine the distribution, mean, standard deviation (SD), range, and the 5th and 95th percentile values. The influence of age, blood pressure, pulse rate, and intraocular pressure on pulsatile ocular blood flow was determined by regression analysis. RESULTS: Reliability coefficient for pulsatile ocular blood flow values ranging from 290 microliters/min to 2,196 microliters/min was 0.92. Variation in bias and first exposure effect were not significant. Pulsatile ocular blood flow values were normally distributed. Mean values were 669.90 +/- 233.0 microliters/min in men and 841.90 +/- 254.6 microliters/min in women. Fifth and ninety-fifth percentile values were 364.75 microliters/min and 1,266.10 microliters/min in men and 397.18 microliters/min and 1,346.10 microliters/min in women. Pulsatile ocular blood flow was significantly influenced by pulse rate. CONCLUSION: This study confirms the reliability of the Ocular Blood Flow Tonograph in repeated measurements of POBF within individuals over short time intervals. The high interindividual variation in POBF may invalidate comparison of POBF between individuals, and the wide range of normal values may limit the value of using a low POBF as a possible indicator of disease. PMID- 9211142 TI - Topical prostaglandins for glaucoma therapy. PMID- 9211143 TI - Current use of ophthalmic beta blockers. PMID- 9211144 TI - Use of antifibrosis agents and glaucoma drainage devices in the American and Japanese Glaucoma Societies. AB - PURPOSE: To investigate practice patterns among glaucoma subspecialists in the American Glaucoma Society (AGS) and the Japanese Glaucoma Society (JGS), regarding use of antifibrosis agents and glaucoma drainage devices. METHODS: An anonymous survey incorporating 10 clinical situations was mailed to all AGS and JGS members in December 1995. RESULTS: Half of the AGS (105 of 210), and JGS (25 of 50) members returned surveys. Most respondents (51-87%) preferred trabeculectomy with adjunctive mitomycin for all 10 clinical situations. Mitomycin concentrations varied from 0.1 to 0.8 mg/ml (range of means for 10 situations 0.31-0.39 mg/ml) and intraoperative application times ranged from 5 s to 7 min (range of means for 10 situations 2.5-4.6 min). Preferences for either no antifibrosis agent (up to 39%) or 5-fluorouracil (up to 29%) were highest in primary trabeculectomy. Thirty-seven percent to 64% of AGS members used glaucoma drainage devices, especially after complicated postsurgical glaucomas (after penetrating keratoplasty, scleral buckling, or pars plana vitrectomy) and in neovascular glaucoma, but few JGS members used them. Large differences between university- and private practice-based AGS members were found only in mitomycin use for primary trabeculectomy (33% vs. 52%, respectively; p = 0.07) and for complicated postsurgical glaucomas (46% vs. 70%, respectively; p = 0.03). CONCLUSIONS: Trabeculectomy with mitomycin was the preferred surgical procedure among AGS and JGS members in the clinical situations surveyed. Mitomycin concentration and time of application varied widely. Many respondents used 5 fluorouracil or no antimetabolite in primary trabeculectomy. Glaucoma drainage devices were widely used for complicated glaucomas in the United States. PMID- 9211145 TI - Glaucomatologists should be certified. PMID- 9211146 TI - Low tension glaucoma. PMID- 9211147 TI - Eye on patients: excerpts from a report on patients' concerns and experiences about the health care system. American Hospital Association and the Picker Institute. AB - The American Hospital Association and The Picker Institute gathered information about patients' perceptions and experiences with health care. The first of a series of reports, Eye on Patients, is excerpted in this article. Preliminary findings show that despite reported rates of "satisfaction" consumers are worried about the future of health care, customer service programs have improved patients' interactions with care givers but these do not address core issues, and patients' experiences reveal problems with the way the system works and how decisions are made about patients' care. PMID- 9211148 TI - How Americans perceive the health care system: a report on a national survey. National Coalition on Health Care. AB - The National Coalition on Health Care telephone poll of a cross section of Americans reveals that most people have little confidence in the health care system. Eight in 10 Americans believe the quality of medical care is being compromised in the interest of profit. Consumers want to be better informed about how to evaluate quality of medical care from physicians and hospitals. The vast majority feels that the federal government can play an important role in improving the health care system. Middle-class respondents were greatly worried about cost, coverage, and treatment. PMID- 9211149 TI - Consumer satisfaction: a key to financial success in the managed care environment. AB - To be successful a company must have the ability to change and be focused on meeting customer needs. The author proposes that health care providers must expand their attention beyond traditional purchasers of services, employers, and insurers and focus on redesigning consumer satisfaction programs and tools to fit the new market environment. Changing from a fee-for-service system to a managed care system means patients are paying an increasing portion of the costs for coverage and care. PMID- 9211150 TI - Preparing for the coming consumer revolution in health care. AB - As consumers shoulder more direct financial responsibility for health care, they will expect health care providers to function like any other consumer service. Health care organizations can prepare for this revolution by (1) benchmarking against world-class companies outside health care; (2) championing benefits, not features; and (3) integrating mortality and morbidity with a product management approach. PMID- 9211151 TI - New HEDIS means more information about HMOs. AB - The latest version of the Health Plan Employer Data and Information Set (HEDIS) is a standardized set of measures to provide health care purchasers and consumers with the information needed to compare the performance of different managed care plans. HEDIS 3.0 combines the last two versions and extends the use to an HMO's Medicare members. The 71 measures are spread over eight topics of interest to purchasers and consumers. PMID- 9211152 TI - Fast, flexible, and fluid: the continuing success of the Charlotte-Mecklenburg Hospital Authority in an era of public hospital crisis. AB - The changing face of health care delivery continues to challenge public hospitals, and many of these hospitals are in danger of closing. Increasing numbers of uninsured patients, coupled with state and federal cuts in Medicaid spending, threaten to worsen the situation. These facilities' survival may well depend upon their ability to create integrated delivery systems (IDSs). However, public hospitals are likely to face significant barriers in forming and participating in IDSs. This article present some of the barriers facing public hospitals as they attempt to form an IDS. Additionally, the authors present a brief case study of a public hospital whose successful efforts to form an IDS began before the IDS concept became popular. In forming an IDS this public hospital has strengthened its commitment to research, education, and the delivery of quality public health care. PMID- 9211153 TI - Hospital and ambulatory surgery center syndications: selling interests to physicians. AB - Physician ownership in hospitals and ambulatory surgery centers remains a relatively surefire method of protecting a portion of a facility's revenues. Implementation of a plan to broaden physician ownership requires compliance with legal and regulatory schemes. This article discusses the prototypical business terms of such transactions, outlines the process for completing such syndications, and analyzes the legal statutes that must be complied with in implementing the effort. PMID- 9211154 TI - A framework for designing and implementing community benefit standards. AB - Increasingly, health care professionals and the public are asking questions about the role of the hospital in meeting community need including its not-for-profit tax status. This article reviews the community benefit literature, provides a framework for understanding how a hospital community benefit program was developed, and delineates through a structured case study the lessons learned from this experience. It provides the practitioner with a context in which other hospitals may replicate the program and gives researchers a substantive case study that may be used as the basis for the empirical testing of community benefit models. The authors also outline the many difficult issues faced by a typical community hospital as it attempted to examine and develop additional responses to community need. PMID- 9211155 TI - Do antibiotic policies have an effect? PMID- 9211156 TI - Serratia marcescens infections in neonatal departments: description of an outbreak and review of the literature. AB - An outbreak of colonization and infection with Serratia marcescens occurred in a neonatal intensive care unit (NICU). S. marcescens was isolated from five preterm infants (gestational age 25-30 weeks). Two infants developed septicaemia, which were both fatal, and one infant (the presumed index case) had conjunctivitis due to S. marcescens. Two infants were colonized without clinical signs of infection. All infants were treated with antibiotic regimens including ciprofloxacin and gentamicin. The DNA fingerprints of isolates were determined by enterobacterial repetitive intergenic consensus primers by the polymerase chain reaction. This showed that a single strain had spread in the NICU. An extensive investigation pointed to an infant born from a mother with an intra-uterine infection after prolonged rupture of foetal membranes as a presumed source of the outbreak. A reservoir, other than the infected or colonized infants and their immediate vicinity, was not found, with the sole exception of the waste jar of a Na+/ K(+) analysis apparatus. Containment of the outbreak was achieved by closure of the NICU for new admissions, strict hygienic measures and cohort nursing of the infected and colonized infants. It was considered especially important to handle the infants with gloves, since frequent hand carriage of staff with S. marcescens was found when gloves were not used. PMID- 9211157 TI - Epidemiologically related and unrelated strains of Pseudomonas aeruginosa serotype O12 cannot be distinguished by phenotypic and genotypic typing. AB - A clonal origin for European isolates of antibiotic multi-resistant Pseudomonas aeruginosa serotype O12 has been suggested. This study was designed to assess the value and limitations of several typing methods for the investigation of outbreaks due to this serotype. In Hopital de Rodez, France, this organism is endemic, and a prospective clinical epidemiological study was undertaken over a 15 month period, encompassing all patients at the hospital from whom P. aeruginosa O12 was isolated. All isolates were examined by auxanogram, antibiogram, phage-typing, electrophoresis of esterases and pulsed-field gel electrophoresis of DNA. The results suggest that (1) the methods used did not clearly differentiate between clinically-related and epidemiologically-unrelated European isolates, (2) in Hopital de Rodez, while some isolates were likely to have been transmitted from patient-to-patient, most infections or colonizations with this organism were sporadic and their origin is unknown. The limits of typing methods for the investigation of outbreaks of nosocomial infection with multi-resistant P. aeruginosa O12 are emphasized. PMID- 9211158 TI - Epidemiology of Pseudomonas aeruginosa in cystic fibrosis and the possible role of contamination by dental equipment. AB - Cystic fibrosis (CF) patients often suffer from Pseudomonas aeruginosa lung infection yet the source of this organism is not known. In order to determine whether CF patients might be contaminated with P. aeruginosa from dental equipment, a total of 103 water samples from 25 dental sessions in Frederiksberg Municipal Oral Health Care Service were examined. Three samples (2.9%) were positive for P. aeruginosa. Three hundred and twenty-seven water samples from 82 dental sessions from various other Municipal Oral Health Services in Denmark, attended by CF patients, were also examined. Eighteen of 327 samples (5.5%) from nine sessions (11%) were positive for P. aeruginosa. In one case, genotypically identical (RFLP, pulsed-field gel electrophoresis) P. aeruginosa strains were found both in water from the dental equipment and in the CF patients sputum. This indicates a small risk for acquiring P. aeruginosa from dental sessions, which is however equal to the yearly 'natural background' incidence (1-2%) of acquisition of P. aeruginosa in our CF centre. PMID- 9211160 TI - Control of varicella-zoster infection on renal and other specialist units. AB - The introduction of chickenpox onto our renal unit recently raised several issues surrounding the management of patient and staff contracts. This paper describes the action taken and makes various recommendations for future management of similar cases. Guidelines are proposed for the management of patients and staff as well as the role of the infection control team in handling a chickenpox problem. Future developments, including the use of VZ vaccine for patient and staff, are also discussed. PMID- 9211159 TI - Application of molecular methods to a nosocomial outbreak of Salmonella enteritidis phage type 4. AB - A nosocomial outbreak of Salmonella enteritidis phage type 4 occurred in July 1995. Seven definite cases were identified over 13 days affecting four wards in a London hospital. The outbreak strain was characterized by plasmid profile typing and pulsed-field gel electrophoresis (PFGE), and was unusual in that it did not possess a 38 MDa plasmid common to most isolates of S. enteritidis PT 4 made from humans and food animals in England and Wales. Seven asymptomatic excreters were identified on screening. No additional cases occurred on wards after standard isolation procedures were implemented. No common or continuing food or dietary source was identified. Results of epidemiological, microbiological and environmental investigations suggested that the outbreak was due to person-to person transmission within the hospital. The source of the outbreak was not established but was probably due to admission of a patient with an unrecognized infection of S. enteritidis PT 4. The study highlights the importance of close collaboration between hospital staff, epidemiologists and microbiologists, and demonstrates the value of molecular techniques for strain subdivision in outbreak investigations. PMID- 9211161 TI - Effect of a commercial disinfectant ('Virkon') on mouse experimental infection by Cryptosporidium parvum. AB - Cryptosporidium parvum oocysts obtained from naturally-infected calves were exposed to 1-10% 'Virkon' for 10-360 min, then inoculated intragastrically into coccidium-free neonatal mice. Prevalence and intensity of infection were determined seven days later by examination of intestinal homogenates. Although we were unable to abolish infectivity for the mice, the intensity of infection was considerably reduced after long periods of exposure (up to > 90%, depending on disinfectant concentration), indicating that this product may have some value for disinfection when extended exposure is possible (e.g., soaking laboratory glassware). PMID- 9211162 TI - Transmission of urinary bacterial strains between patients with indwelling catheters--nursing in the same room and in separate rooms compared. AB - Despite lack of supporting scientific data it has been suggested that patients with an indwelling urinary catheter (IUC) should be nursed in separate rooms to reduce the risk of cross-infection. We conducted a one-month case-control study of nursing home patients with an IUC and bacteriuria, 20 nursed together pairwise and 20 in separate rooms, by weekly urine cultures and typing of the bacterial isolates. The transmission rate of urinary strains between patients was three times higher within rooms (5/9 possible transmissions) than between rooms (9/53 possible transmissions, P = 0.02). The study thus supported nursing IUC patients in separate rooms. PMID- 9211163 TI - Management of an outbreak of methicillin-resistant Staphylococcus aureus in a risk area with empirical intranasal mupirocin. PMID- 9211164 TI - Do patients with Clostridium difficile need to be isolated? PMID- 9211165 TI - A bacteriological study of a contamination control tacky mat. PMID- 9211166 TI - Nosocomial infections in emergency units of Brazilian hospitals. PMID- 9211167 TI - Debate: vascular remodelling. PMID- 9211168 TI - Long-term inhibition of the renin-angiotensin system in genetic hypertension: analysis of the impact on blood pressure and cardiovascular structural changes. AB - OBJECTIVE: To compare, using data from published studies, the efficacy of chronic inhibition of the renin-angiotensin system in inducing persistent downregulation of hemodynamic and cardiovascular structural changes in an adult rat with established genetic hypertension with the widely accepted known downregulation in young genetically hypertensive rats. STUDY SELECTION: We report on 36 studies that satisfied our inclusion criteria (angiotensin converting enzyme inhibitor or angiotensin II receptor antagonist treatment that lowered arterial pressure levels for at least 3 weeks). Of the 24 studies concerning developing hypertensive rats, a significant number (n = 17) also examined the persistence of any hemodynamic or cardiovascular effects after withdrawal of treatment. Conversely, of 15 studies using adult rats only seven and three reported on post treatment hemodynamic and cardiovascular structural indices respectively. RESULTS: During treatment the hemodynamic and cardiovascular structural changes produced were qualitatively and quantitatively similar in the young and adult treated rats. Critical assessment of the persistence of these effects after withdrawal of treatment again found qualitatively similar responses. However, the strength of this finding is limited by the paucity of studies concerning adult rats in which equivalent treatment durations and equipressor doses of treatments were compared between these two age groups. CONCLUSIONS: Blockade of the renin angiotensin system appears to have an efficacy in reversing established hypertension and hypertrophy similar to that with which it prevents the development of hypertension and hypertrophy. This partial 'cure' of hypertension after withdrawal of treatment is clearly evident when treatment is initiated during the development of hypertension and appears to be similar even when treatment is initiated in established hypertension. PMID- 9211169 TI - Increased levels of tissue plasminogen activator antigen in essential hypertension. A population-based study in Sweden. AB - OBJECTIVE: To investigate components of the haemostatic and fibrinolytic system in borderline hypertensives and hypertensives, drug-treated or not, from a defined population. DESIGN AND METHODS: A randomly selected sample of the population of northern Sweden, 1558 subjects aged 25-64 years, was studied. Eight per cent of them were being treated with antihypertensive drugs (trHT). Remaining subjects were classified according to their mean diastolic blood pressure (DBP). Normotension, DBP < 85 mmHg, was found in 63%, borderline hypertension (bHT), DBP 85-94 mmHg, in 21% and untreated hypertension (uHT), DBP > or = 95 mmHg, in 8% of the subjects. RESULTS: Mean age increased from the normotensive group through the bHT and uHT groups to the trHT group, members of which were the oldest. Age adjusted values for the body mass index, waist: hip ratio, serum triglyceride and Phadeseph plasma insulin levels increased with each level of hypertension. Plasma fibrinogen levels and plasminogen activator inhibitor type 1 activity (in men) increased stepwise from normotensives through bHT and uHT to the highest values found in the trHT group. The tissue plasminogen activator (tPA) activity in men declined strongly across the groups, trHT having the lowest fibrinolytic activity (P < 0.001). tPA antigen levels increased strongly from normotensives through bHT to uHT, but then were lower in the trHT group. Even after adjustment for possible confounders, men in the uHT group had 21% higher (P = 0.027) tPA antigen levels than did the normotensives. In bHT men, the tPA antigen and plasminogen activator inhibitor type 1 activities were 14 and 24% respectively, higher (P < 0.01) than those in the normotensives. CONCLUSION: Hypertension is associated with multiple metabolic and fibrinolytic disturbances that are accentuated in drug-treated hypertensives and already discernible in subjects with borderline hypertension. Decreased fibrinolysis is associated with, and possibly secondary to, metabolic disturbances linked to the insulin-resistance syndrome. The independent increase in tPA antigen in hypertensive men might indicate an endothelial dysfunction. PMID- 9211170 TI - Prediction of mortality by ambulatory blood pressure monitoring versus screening blood pressure measurements: a pilot study in Ohasama. AB - OBJECTIVE: To compare the prediction of mortality by ambulatory blood pressure monitoring and screening blood pressure measurements in a general population. DESIGN: A prospective cohort study. PATIENTS AND METHODS: We obtained blood pressure data for 1542 subjects (565 men and 977 women) aged > or = 40 years who were followed up for up to 8.1 years (mean 5.1 years). Subjects were subdivided into five groups according to their ambulatory and screening blood pressure levels. The prognostic significance of blood pressure for mortality was examined by the Cox proportional hazards regression model. RESULTS: The association between blood pressure level and mortality was more distinctive for the ambulatory blood pressure than it was for the screening blood pressure. The risk of cardiovascular mortality increased significantly for the highest quintiles of 24 h ambulatory blood pressure, whereas there was no significant association between the screening blood pressure and the cardiovascular mortality. When both 24 h and screening blood pressure values were included in the Cox model, only the systolic ambulatory blood pressure was related significantly to the increased risk of cardiovascular mortality. CONCLUSIONS: The ambulatory blood pressure had a stronger predictive power for mortality than did the screening blood pressure. This appears to have been the first study of the prognostic significance of ambulatory blood pressure monitoring versus screening blood pressure measurements in a general population. PMID- 9211171 TI - Analysis of phenotypic consequences of renin gene polymorphism in Lyon rats. AB - OBJECTIVE: To investigate phenotypic consequences of renin gene polymorphism between Lyon hypertensive (LH) and normotensive (LN) rats because previously we demonstrated cosegregation of the LH allele with increased blood pressure in a cross of LH with LN rats. DESIGN: Two studies were conducted. Study 1 used a cohort of male F2 rats from a LH x LN cross. Eighty-two rats homozygous for the hypertensive (HH) renin gene allele were compared with 82 rats homozygous for the normotensive (NN) allele. Urinary steroid excretion was measured in 24 h urine samples collected from rats aged 6 weeks. The direct aortic blood pressure was recorded in 30-week-old rats and, after they had been killed, their kidney renin concentration (KRC) was measured. In study 2, renin, angiotensinogen and angiotensin converting enzyme plasma concentrations and renin messenger RNA (mRNA) levels were measured in renal and extra-renal tissues from 6- and 25-week old LH and LN parental and HH and NN F2 male rats. METHODS: Urinary steroids and plasma components of the renin-angiotensin system (RAS) were measured using specific radioimmunoassays. mRNA levels were quantified by northern blotting. RESULTS: In study 1, HH F2 rats had a higher blood pressure (151.5 +/- 8.2 versus 146.0 +/- 7.4 mmHg, P < 0.001) and a lower KRC (514 +/- 203 versus 666 +/- 304 micrograms A1/h per g cortex, P < 0.01) than did NN rats aged 30 weeks. In covariate analysis the decrease in KRC in HH rats was attributable to their increased blood pressure rather than to the renin genotype. The renin genotype of rats aged 6 weeks was not associated with a change in the urinary excretion of aldosterone, desoxycorticosterone, corticosterone or 18-hydroxy desoxycorticosterone. In study 2, we found no difference either in plasma levels of RAS components or in renal or extrarenal renin mRNA levels either between parental LH and LN rats or between HH and NN F2 rats apart from a higher plasma renin concentration in LH rats aged 6 weeks. Renal, but not extra-renal, renin mRNA levels declined with age. CONCLUSIONS: We found no evidence of a renin genotype-dependent phenotypic difference in the RAS that could account for the effect of the renin locus on blood pressure in Lyon rats. Our findings suggest that the effect of the locus on blood pressure might be due to an as yet unidentified gene linked to renin. PMID- 9211172 TI - Chronic blockade of nitric oxide synthesis increases urinary endothelin-1 excretion. AB - OBJECTIVES: Our objective was to determine the effect of nitric oxide (NO) inhibition on renal synthesis of endothelin-1 (ET-1) in vivo. DESIGN AND METHODS: Rats were administered 500 mg/l NG-nitro-L-arginine methyl ester (L-NAME) in their drinking water or its vehicle for 2 weeks (2W-L-NAME, n = 10; 2W-CONT, n = 10) or for 6 weeks (6W-L-NAME, n = 13; 6W-CONT, n = 11). We measured the levels of albumin, NO metabolites and ET-1 both in their blood and in 24 h urine samples, and determined the expression of preproET-1 messenger RNA in the renal cortex and the inner medulla. We also examined renal histology. RESULTS: L-NAME administration for 6 weeks reduced NO metabolites both in serum (21.5 versus 3.66 nmol/ml in 6W-CONT) and in urine (5.72 versus 22.53 nmol/24 h in 6W-CONT), raised the systolic blood pressure (228 versus 162 mmHg in 6W-CONT), and the increased urinary excretion of albumin (24.29 +/- 11.66 versus 0.60 +/- 0.08 mg/day in 6W CONT) and of ET-1 (112.0 +/- 38.3 versus 35.8 +/- 4.4 pg/day in 6W-CONT). There were no significant differences between the plasma levels of ET-1 in the control and L-NAME groups. Expression of preproET-1 messenger RNA increased in the renal cortex but not in the inner medulla in the 6W-L-NAME group. Bleeding and marked arteriolar narrowing were observed in the renal cortex of the 6W-L-NAME group. CONCLUSIONS: Prolonged inhibition of NO synthesis increases urinary excretion of ET-1 and albumin without having any effect on plasma ET-1 levels. These results do not support the hypothesis that NO plays an inhibitory role in the regulation of ET-1 in the systemic circulation, although it is possible that such a role could exist in renal tissue. However, in view of the albuminuria, a more likely explanation is that increased urinary ET-1 is secondary to L-NAME-induced renal hyperfiltration injury. PMID- 9211173 TI - Cold-induced stress stimulates the sympathetic nervous system, causing hypertension and proteinuria in rats. AB - OBJECTIVE: To determine whether cold-stress stimulation of the soles of the paws would produce a preeclampsia-like syndrome in rats. METHODS: Pregnant or nonpregnant rats were kept in 0 degree C floor and 23 degrees C room temperature cages (the cold-stressed group) or in 23 degrees C floor and 23 degrees C room temperature cages (the control group) for 2 weeks. Their blood pressure, proteinuria, and plasma catecholamines were measured, and histologic studies were performed on all groups. RESULTS: There were no significant differences in systolic blood pressure between the two groups during the first week of the experimental period; however, during the last week of gestation the blood pressure of the cold-stressed group did not fall and was significantly higher than that of the control group. A significant increase in urinary protein excretion was observed in the cold-stimulated pregnant rats, in contrast to the control rats. The concentrations of norepinephrine and epinephrine in the cold stressed pregnant rats were markedly higher than those in the control rats. A decrease in trophoblast invasion, congestion, and fibrinoid deposits of the labyrinth were observed in the cold-stressed rats. A marked increase in subendothelial fibrinoid deposits in the glomerular capillary was found only in the cold-stressed pregnant rats. The blood pressure, biochemical parameters, and histologic findings in the nonpregnant rats were almost the same as those in the pregnant rats. CONCLUSION: Chronic local cold stimulation of the soles of the paws induces preeclampsia-like phenomena in pregnant and nonpregnant rats, and this model suggests that the cause of preeclampsia is involved in chronic stimulation of the sympathetic nerve. PMID- 9211174 TI - Validation of a continuous baroreceptor reflex sensitivity index calculated from spontaneous fluctuations of blood pressure and pulse interval in rats. AB - OBJECTIVE: To develop and validate a technique for the continuous computerized calculation of the baroreceptor reflex sensitivity (BRS) of the heart rate in rats. DESIGN: The BRS was calculated from spontaneous changes in blood pressure and pulse interval using spectral analysis as well as time-series techniques. The BRS values obtained with these techniques were compared with those obtained by standard pharmacological methods. METHODS: The blood pressure and pulse interval in adult Wistar-Kyoto (WKY) rats were recorded on a beat-to-beat basis for two consecutive 30 min periods. During one of these periods the BRS was determined pharmacologically by injections of nitroprusside and phenylephrine. Measurements were performed after administration of saline as vehicle or during manipulation of the autonomic nervous system by infusion of metoprolol, methyl-atropine and hexamethonium. Sequential time-series methods for continuous BRS calculation were tested for 24 h periods in intact WKY rats as well as in WKY rats that had been subjected to sino-aortic denervation or to electrical lesioning of the nucleus tractus solitarius. RESULTS: The correlation coefficient between BRS values in intact WKY rats derived from the pharmacological method and those from spectral analysis techniques was low (R2 = 0.16). The correlation coefficient between BRS values from the pharmacological method and those from the developed time-series method was higher (R2 = 0.64). The BRS measured using the latter method was found to vary over 24 h with the highest values during the sleeping period. After surgical elimination of the baroreflex, the algorithm returned BRS values close to zero throughout the 24 h period. The BRS estimate was found to be a measure of the parasympathetic rather than of the sympathetic component of the baroreceptor reflex. CONCLUSION: The developed time-series method calculates an index of the gain of the cardiac baroreflex in rats faithfully. This method can be implemented in data acquisition software, allowing continuous on-line monitoring of the cardiac baroreflex gain. PMID- 9211175 TI - Autonomic control of ultradian and circadian rhythms of blood pressure, heart rate, and baroreflex sensitivity in spontaneously hypertensive rats. AB - OBJECTIVE: To examine the influence of the autonomic nervous system on ultradian and circadian rhythms of blood pressure, heart rate and baroreflex sensitivity of heart rate (BRS) in spontaneously hypertensive rats (SHR). METHODS: Spontaneous fluctuations in blood pressure, heart rate and BRS in SHR were recorded continuously for 24 h using a computerized system and compared with those in Wistar-Kyoto (WKY) rats. Furthermore, 24 h recordings were performed in SHR during cardiac autonomic blockade by metoprolol and methyl-atropine, vascular autonomic blockade by prazosin, ganglionic blockade by hexamethonium and vagal stimulation by a low dose of scopolamine. The magnitudes of the ultradian fluctuations in blood pressure, heart rate and BRS were assessed by wide-band spectral analysis techniques. RESULTS: The BRS was lower in SHR than it was in WKY rats throughout the 24 h cycle. In both strains high values were found during the light, resting period, whereas low values were found during the first hours of the dark, active period. The circadian rhythmicity of the blood pressure in SHR was abolished completely during the infusions of prazosin and hexamethonium. In contrast, the circadian rhythmicities of the blood pressure and heart rate were not altered by infusions of metoprolol, methyl-atropine and the low dose of scopolamine. Power spectra of the blood pressure and heart rate lacked predominant peaks at ultradian frequencies and showed 1/f characteristics. In the absence of autonomic tone, the ultradian fluctuations in heart rate, but not in blood pressure, were decreased. The ultradian BRS spectra had no 1/f shape, but showed a major peak at approximately equal to 20 min for 71% of the WKY rats and 42% of the SHR. CONCLUSIONS: The influence of the autonomic nervous system on the blood pressure and heart rats in SHR is frequency-dependent. The circadian, but not ultradian, blood pressure rhythmicity is controlled by vascular autonomic activity. Conversely, the circadian, but not ultradian, heart rate rhythmicity is independent of autonomic tone. In rats, just as in humans, the trough in baroreflex sensitivity occurred after the sleeping period, when locomotor activity is resumed. PMID- 9211176 TI - Arteriolar constriction in mild-to-moderate essential hypertension: an old concept requiring reconsideration? AB - OBJECTIVE: To investigate differences between in-vivo properties of a vascular bed in hypertensive patients and normotensive controls. DESIGN: Despite the controversy about the origin of essential hypertension and its accompanying vascular changes, it is generally assumed that the characteristic increase in peripheral resistance when hypertension progresses is caused by arteriolar constriction. Yet, there is little experimental evidence that this assumption generally holds in vivo. METHODS: A non-invasive technique was used for studying properties of the complete vascular bed of an upper arm segment under an occluding cuff in 23 previously untreated hypertensive patients and their matched normotensive controls. The method used the segment's electrical impedance to assess the volumes of extravascular fluid and of arterial and venous blood under varying arterial transmural pressures. RESULTS: Compared with that of matched normotensive controls, the compliance of the large arteries of the vascular bed was on average 50.9% lower (P < 0.001) in the hypertensive patients. The compliance of the complete arterial bed at the operating blood pressure level was also lower (40.0%, P < 0.01), but appeared to be significantly higher (45.9%, P < 0.05) at the normotensive blood pressure level. On the venous side, the patients had a higher blood volume (60.0%, P < 0.01) and an increased myogenic response (68.5%, P < 0.05). CONCLUSIONS: The increase in vascular resistance in the hypertensive patients is due primarily to changes in the large and small vessels of the arterial bed. We found no evidence for a generally increased arteriolar constriction. PMID- 9211177 TI - Kinins and the events influenced by an angiotensin-converting enzyme inhibitor during neointima formation in the rat carotid artery. AB - OBJECTIVE: In balloon-injured rat carotid arteries, angiotensin-converting enzyme inhibitors (ACEI) decrease neointima formation, and a kinin receptor antagonist partially reverses this inhibitory effect. We studied which of the events leading to neointima formation are involved in the effects of ACEI and kinins. METHODS: We administered 5 mg/kg per day ramipril, either alone or combined with the kinin receptor antagonist icatibant (Hoe 140), on the days each wave occurred and studied the effects on neointima formation 14 days after balloon injury. Ramipril alone or combined with icatibant had no effect on neointima formation when administered from 2 days before to 3 or 5 days after balloon injury. In contrast, ramipril inhibited neointima formation when administered from day 7 to day 14. Treatment with icatibant had a small effect, which was sufficient to abolish the effects of ramipril (control 0.11 +/- 0.01 mm2, ramipril 0.08 +/- 0.01 mm2; P < 0.05; ramipril plus icatibant 0.09 +/- 0.01 mm2; NS, ramipril plus icatibant versus control). Thus ramipril was not effective when treatment was stopped after 3 or 5 days, but was mildly effective when treatment was administered during the second week. The effect on migration was studied by counting the number of neointimal cells in rats treated from 2 days before to 4 days after injury. Ramipril decreased the number of cells by 93% compared with controls (control 65.0 +/- 13.5 cells/slice, ramipril 4.7 +/- 2.0 cells/slice; P < 0.001), and this effect was blunted significantly by icatibant (19.5 +/- 5.7 cells/slice; P < 0.009, versus ramipril; P < 0.007, versus controls). The influence of treatment on the rate of proliferation (the 5'-bromo-2'-deoxyuridine index) was studied in the media 3 days, and in the neointima 7 and 10 days after balloon injury. Although proliferation peaked in the neointima after 7 days, there were no differences among the groups at any time. Thus neither ramipril nor icatibant affected the rate of proliferation at the times sampled. Ramipril increased cell density (cells/mm2) in the neointima, and this effect was abolished by cotreatment with icatibant (P < 0.05). CONCLUSION: The ACEI needs to be present throughout the experimental period to be most effective. ACEI act on neointima formation in part by inhibiting migration; thus, because ramipril was mildly effective when administered from 7 to 10 days after injury, it is likely that vascular smooth muscle cell migration also occurs continuously. Kinins help mediate roughly 30% of the effect of ACEI on migration. In addition, ACEI, through kinins, affect a process that increases the density of the cells in the neointima, perhaps by decreasing extracellular matrix deposition. PMID- 9211178 TI - Activation of angiotensin II-forming chymase in the cardiomyopathic hamster heart. AB - BACKGROUND: Angiotensin (ANG) II plays crucial roles in promoting cardiovascular tissue remodeling. Human chymase catalyzes ANG II formation, whereas rat chymase (rat mast cell protease 1) degrades ANG I to inactive fragments. Such species differences should be considered when the functions of chymase in human cardiovascular diseases are investigated assuming an analogy with animal models. OBJECTIVE: To further characterize the recently identified ANG II-forming hamster chymase, and to analyze pathophysiologic roles played by chymase in the cardiomyopathy of the hamster. METHODS: The gene organization and the primary structure of hamster chymase were determined through molecular cloning. Chymase and angiotensin converting enzyme messenger RNA levels, and chymase-like and angiotensin converting enzyme activities were measured in the heart of BIO 14.6 cardiomyopathic hamsters aged 4, 12, and 25 weeks. RESULTS: The hamster chymase gene is 3 kb long. It has five exons and four introns, and the deduced amino-acid sequence was homologous to other mammalian chymases. The chymase messenger RNA levels and chymase-like activities in the BIO 14.6 hamster hearts were increased significantly at the ages of 12 weeks (the fibrotic stage) and 25 weeks (the hypertrophic stage), but not at age 4 weeks (the premyolytic stage). CONCLUSIONS: These results indicate that heart chymase is activated concurrently with the development of cardiomyopathy. Thus, we conclude that heart chymase could play the primary role in accelerating ANG II formation, thereby causing deleterious changes in the cardiomyopathic heart. PMID- 9211179 TI - Progression of renal failure after subtotal nephrectomy in transgenic rats carrying an additional renin gene [TGR(mREN2)27]. AB - OBJECTIVE: To study the evolution of glomerulosclerosis after renal ablation in a model with abnormal regulation of the renin gene. METHODS: Four-month-old female ovariectomized hypertensive heterozygous transgenic rats (TGR) harbouring the murine REN-2 gene were compared with pressure-matched, pair-fed, stroke-prone, spontaneously hypertensive rats (SHRsp). Both groups were followed for 6 weeks after 70% subtotal nephrectomy (SNX) or sham operation. RESULTS: Blood pressures in the SNX group at the end of the experiment were 193 +/- 3 mmHg in TGR and 199 +/- 5 mmHg in SHRsp. The final C(in) was 306 +/- 68 microliters/min per 100 g body weight in TGR that had undergone SNX and 550 +/- 93 microliters/min per 100 g body weight in SHR that had undergone SNX (P < 0.02), whereas inulin clearance (C(in)) in sham-operated pair-fed TGR and SHRsp controls did not differ from each other. The glomerulosclerosis index was 1.75 +/- 0.08 in perfusion-fixed TGR that had undergone SNX versus 1.21 +/- 0.03 in SHR that had undergone SNX (P < 0.005). In addition, the media thickness of preglomerular vessels was significantly greater in TGR that had undergone SNX (7.48 +/- 0.79 microns) than it was in SHRsp that had undergone SNX (5.27 +/- 1.38 microns, P < 0.02). Rat renal renin messenger RNA (mRNA) expression and, in parallel, mouse REN-2 gene expression were lower in TGR after SNX. Plasma renin and angiotensin II (ANG II) concentrations were reduced to a similar extent in both SNX groups, but plasma prorenin was higher in TGR that had undergone SNX than it was in SHRsp that had undergone SNX. The angiotensin II:I ratio in the kidney was significantly higher in TGR (P < 0.01). There was no significant difference between sham-operated or subtotally nephrectomized TGR and SHRsp with respect to angiotensin type 1 mRNA and angiotensinogen mRNA. The renal angiotensin converting enzyme activity, however, was significantly higher in sham operated and subtotally nephrectomized TGR than it was in sham operated SHRsp and in SHRsp that had undergone SNX. CONCLUSION: Deterioration of renal function is accelerated in subtotally nephrectomized transgenic rats [TGR(mREN2)27] compared with that in comparably hypertensive SHRsp despite suppressed circulating active mRNA and decreased renal renin mRNA. Although alternative explanations are possible, this observation is consistent with a role for local ANG II in the genesis of glomerulosclerosis. PMID- 9211180 TI - History of hypertension in patients treated for end-stage renal disease. AB - OBJECTIVES: To describe patterns of hypertension history in patients with various types of end-stage renal disease (ESRD) and in persons with normal kidney function; and to identify risk factors for the diagnosis 'hypertensive ESRD'. DESIGN: A case-control study. SETTING: Population-based. PARTICIPANTS: Patients with ESRD due to hypertension (n = 214), diabetes (n = 239), other specified causes (n = 181), unknown causes (n = 82) and control subjects drawn from the general population (n = 361). MAIN OUTCOME MEASURES: Participants' history of hypertension. RESULTS: The prevalence of hypertension was 90% in ESRD patients and 27% in controls. Only 6% of patients with hypertensive ESRD had a history of malignant hypertension. Patients with hypertensive ESRD were more likely to have been hospitalized because of hypertension (36%) than were other ESRD patients (18%) or controls (5%). ESRD of any cause was more strongly associated with hypertension of > or = 25 years duration (odds ratio 51.0, compared with normal blood pressure) than it was with hypertension of shorter duration (15-25 years: odds ratio 31.8, 5-15 years: odds ratio 16.0, < 5 years: odds ratio 21.2). Among patients who had both hypertension and ESRD, the diagnosis of 'hypertensive ESRD' was associated independently with a long duration of hypertension, greater severity of hypertension, the absence of diabetes, black race, and limited education. CONCLUSIONS: Hypertension is common among patients with ESRD. The risk of ESRD from any cause increases progressively with the duration of hypertension, and with indicators of severe hypertension. This result supports the hypothesis that nonmalignant hypertension of long duration may cause renal insufficiency. The criteria used to diagnose hypertensive ESRD are consistent with pathophysiologic and epidemiologic evidence. PMID- 9211181 TI - Clinical versus experimental utility of salt-sensitivity testing. PMID- 9211182 TI - Salt, resting arterial pressure and 24 h monitoring. PMID- 9211183 TI - Inborn errors of signal transduction: mutations in G proteins and G protein coupled receptors as a cause of disease. AB - A vast array of neurotransmitters, polypeptide hormones and other extracellular signalling molecules utilize G protein-coupled pathways for transmembrane signalling. In recent years, mutations in G protein-coupled receptors and in G protein alpha subunits have been identified as the cause of a variety of human diseases. Both loss and gain of function mutations have been described in disorders such as Albright hereditary osteodystrophy, nephrogenic diabetes insipidus, McCune-Albright syndrome, and familial male precocious puberty. Identification of mutations in G protein-coupled receptors and in G proteins in human diseases has provided unique insights into G protein-coupled signal transduction, has important implications for diagnosis and potentially for treatment, and should stimulate the search for additional defects in G protein coupled signal transduction in other diseases. PMID- 9211184 TI - Trinucleotide repeat disorders. PMID- 9211187 TI - Kennedy disease. AB - Kennedy disease is a disorder with progressive motor neuron degeneration that is caused by trinucleotide repeat expansion in the androgen receptor gene. The disease mechanism likely involves toxicity of an expanded polyglutamine tract in the androgen receptor protein. This mechanism is probably shared by other neurodegenerative disorders with polyglutamine expansion, including Huntington disease. Attempts at reproducing the Kennedy disease phenotype by introducing the expanded androgen receptor into cultured neuronal cells and transgenic animals have thus far been unsuccessful, but recently developed model systems with other expanded polyglutamine constructs should allow the pathogenesis of these diseases to be elucidated. PMID- 9211185 TI - Huntington disease: advances in molecular and cell biology. AB - Huntington disease is an inherited neurodegeneration, for which the associated mutation was isolated in 1993. The mutation is an expansion of a CAG trinucleotide repeat, which translates to give a polyglutamine tract at the N terminus of a large protein, huntingtin. Neither the normal nor the pathogenic functions of this protein have been identified, but it is clear that pathogenesis is mediated through the expanded polyglutamine tract within the protein, and that polyglutamine is toxic to cells. A number of proteins which interact with the N terminal region of huntingtin have been isolated, but this has not, so far, yielded a rationale for pathogenesis. Huntingtin is found in areas of the brain that degenerate in this disease but is also associated with pathogenic inclusions in Alzheimer disease and Pick disease. It is possible that Huntington disease has pathogenic mechanisms in common with these other neurodegenerative diseases, and that the mechanism may relate to the formation of abnormal, cytoskeletal associated, inclusions within cells. PMID- 9211186 TI - The fragile X syndrome. AB - The fragile X syndrome is caused by the amplification of a polymorphic CGG repeat in the 5' untranslated region of the FMR1 gene and is the most common form of inherited mental retardation. When the repeat is amplified beyond 200 repeat units, the repeat and the FMR1 promoter region are methylated. As a result of this methylation the gene is silenced and no FMR1 gene product (FMRP) is translated. The lack of expression of FMRP in the fragile X syndrome causes the fragile X phenotype. A mouse model for the fragile X syndrome (knockout for FMRP) has been generated to study the pathological mechanisms leading to the symptoms seen in fragile X patients. FMRP is widely expressed in different tissues and localized predominantly in the cytoplasm associated with the 60S ribosomal subunit. The protein has RNA binding properties and possibly shuttles between cytoplasm and nucleus. The target signals necessary for this intracellular transport, like a nuclear location signal and a nuclear export signal, are present in FMRP. FMRP is also able to bind to other proteins by using specific sequence domains present in the protein. The coiled-coil structures formed by these domains are known to be involved in protein-protein interaction. In this review we postulate that FMRP is involved in the transport of RNA and/or proteins from the nucleus to the cytoplasm. PMID- 9211188 TI - Myotonic dystrophy: molecular and cellular consequences of expanded DNA repeats are elusive. AB - The mutation in the myotonic dystrophy (DM) gene is an expansion in a triplet (CTG) repeat in the 3' untranslated region of a novel gene that partially encodes a serine-threonine protein kinase (DMPK), with closest sequence homology to a small subgroup of protein kinases involved in the control of proliferation and cell shape. Expansion of the repeat correlates reasonably well with disease severity and offers a plausible molecular explanation for the previously contentious issue of anticipation. There is considerable heterogeneity in CTG expansion size in different tissues of affected individuals. The consensus of data from many laboratories indicates that DMPK mRNA is most probably downregulated as a consequence of the repeat expansion. Two polypeptides (68/78 kDa) have been shown to be absent in mouse knockout mutants and therefore can be considered as bona fide gene products. Previous data suggesting that 52-55 kDa polypeptides were likely candidates, have been firmly ruled out at the same time. Further results from studies of knockout and overexpressing transgenic mice indicate that neither simple loss nor gain of DMPK expression is sufficient to account for the DM clinical phenotype. One of the most pressing questions now being addressed is how expansion of the CTG repeat within the DMPK gene affects gene expression, not only of DMPK, but of all genes at the 19q13.3 locus: is DMPK actually responsible for the clinical phenotype seen in DM? The identification of both immediate upstream and downstream human genes (59 and DMRHP, respectively) has been an important first step to answering these questions. Only when these matters have been dealt with can one reasonably expect to start to delineate the different metabolic and signalling pathways responsible for the diverse phenotypes that make up the complex clinical picture of DM. PMID- 9211189 TI - The recognition of Lesch-Nyhan syndrome as an inborn error of purine metabolism. AB - Lesch-Nyhan syndrome was first described over thirty years ago. The original patient was a 4-year-old boy with neurological abnormalities as well as haematuria. Crystals in his urine were identified and confirmed to be uric acid. The massive excretion of this purine led to metabolic studies using isotopically labelled uric acid to study turnover rates. Clues to the site of the enzyme defect resulted from studies with the immunosuppressive agent azathioprine, which normally causes uric acid concentrations to fall in blood and urine but was without effect in a Lesch-Nyhan patient. A deficiency of hypoxanthine phosphoribosyltransferase (HPRT) activity explained this observation in Lesch Nyhan patients. Subsequent studies have indicated that the degree of HPRT deficiency appears to determine the severity of the disease. Molecular studies have shown that most families carry a unique mutation. Attempts are being made to correlate the type and site of a specific mutation with a particular phenotype. PMID- 9211191 TI - Clinical heterogeneity and molecular mechanisms in inborn muscle AMP deaminase deficiency. AB - Lack of the muscle-specific isoform of AMP deaminase (myoadenylate deaminase deficiency) can cause a metabolic myopathy, with exercise-induced muscle symptoms such as early fatigue, cramps and/or myalgia. It is the most common muscle enzyme defect in man, found in about 2-3% of all muscle biopsies. The genetic basis of the inherited defect is the nonsense mutation C34-T in the AMPD1 gene encoding myoadenylate deaminase. The mutation results in a premature stop of the enzyme synthesis. In a healthy German population, the frequency of the mutant allele was 0.1, and 1% of this population is expected to be homozygous for the mutation. In people with muscle symptoms, the allele frequency was significantly higher (0.145). The correlation between allele frequency and muscle symptoms underscores the clinical significance of this defect. However, the vast majority of homozygous subjects do not develop a metabolic myopathy. This clinical heterogeneity may be due to molecular genetic factors such as alternative splicing of the exon harbouring the mutation, or due to metabolic conditions such as pathways compensating for the defect. The real basis for the high percentage of asymptomatic homozygous subjects remains to be revealed. PMID- 9211190 TI - Clinical expression, genetics and therapy of adenosine deaminase (ADA) deficiency. AB - Adenosine deaminase (ADA) deficiency was the first known cause of primary immunodeficiency. Over the past 25 years the basis for immune deficiency has largely been established. Now it appears that ADA deficiency may also cause hepatic toxicity, raising new questions about its pathogenesis. The ADA gene has been sequenced and the ADA three-dimensional structure solved. The relationship between genotype and phenotype is being analysed, and ADA deficiency has become a focus for novel approaches to enzyme replacement and gene therapy. PMID- 9211192 TI - Inborn errors of the purine nucleotide cycle: adenylosuccinase deficiency. AB - Adenylosuccinase catalyses two reactions in purine metabolism: the conversion of succinylaminoimidazole carboxamide ribotide (SAICAR) into aminoimidazole carboxamide ribotide (AICAR) along the de novo synthesis of purine nucleotides, and the conversion of adenylosuccinate (S-AMP) into AMP in the conversion of IMP into AMP. The hallmarks of adenylosuccinase deficiency are the presence of succinylaminoimidazole carboxamide riboside (SAICAriboside) and succinyladenosine (S-Ado) in body fluids. These normally undetectable succinylpurines are the products of the dephosphorylation, by cytosolic 5'-nucleotidase, of the two substrates of adenylosuccinase. The clinical picture of the enzyme deficiency is markedly heterogeneous with, as a rule, a profound, but nevertheless variable degree of psychomotor delay, often convulsions and/or autistic features, sometimes growth retardation and muscular dystrophy. The diagnostic tests that can be used for diagnosis, the enzyme and gene defects that have been identified, and the hypotheses that have been put forward to explain the pathophysiology of the disorder are reviewed. PMID- 9211193 TI - Inborn errors of pyrimidine degradation: clinical, biochemical and molecular aspects. AB - The pyrimidines, uracil and thymine, are degraded in four steps. The first three steps of pyrimidine catabolism, controlled by enzyme shared by both pathways, result in the production of the neurotransmitter amino acid beta-alanine from uracil and the nonfunctional (R)-(-)-beta-aminoisobutyrate from thymine. The fourth step is controlled by several aminotransferases, which have different affinities for beta-alanine, beta-aminoisobutyrate and GABA. Defects concerning the first three steps all lead to a reduced production of beta-alanine; defects of the transaminases involving the metabolism of beta-alanine and GABA lead to accumulation of these neurotransmitter substances. In addition, other metabolites will accumulate or be reduced depending on the specific enzyme defect. Analysis of the abnormal concentrations of these metabolites in the body fluids is essential for the detection of patients with pyrimidine degradation defects. Clinically these disorders are often overlooked because symptomatology is highly aspecific. The growth in our knowledge concerning inborn errors of pyrimidine degradation has emphasized the importance of the clinical awareness of these defects as a possible cause of neurological disease and a contraindication for treatment of cancer patients with certain pyrimidine analogues. The various defects are discussed and attention is paid to clinical genetic and diagnostic aspects. PMID- 9211194 TI - When to investigate for purine and pyrimidine disorders. Introduction and review of clinical and laboratory indications. AB - When to suspect and thus investigate for inborn errors of purine and pyrimidine metabolism is a dilemma for even the most observant investigator. Often parents of affected children, or a history involving siblings, can provide valuable clues. The recognition of new purine and pyrimidine disorders requires skill and serendipity. But even identifying known disorders can prove difficult, since they cover a broad spectrum of illnesses, can have more than one symptom, or lead to early death. This problem is compounded by the fact that they are relatively recently described and therefore often little known, either in the clinic or laboratory. The considerable heterogeneity in clinical expression within families as well as between families means that asymptomatic homozygotes may not be recognized or can present at any time from early childhood through adolescence up to their eighth decade. Consequently, all siblings should be screened. These disorders should be suspected in any case of unexplained anaemia, failure to thrive, susceptibility to recurrent infection, or neurological deficits with no current diagnosis, including autism, cerebral palsy, delayed development, deafness, epilepsy, self-mutilation, muscle weakness, the inability to walk or talk, and-unusual in children and adolescents-gout, sometimes with renal disease. Some disorders present with radiolucent kidney stones, in acute or chronic renal failure, alone or with any of the above, or as an intolerance/sensitivity to therapy (e.g. 5-fluorouracil in malignancies or azathioprine immunosuppression in organ transplantation), often with life-threatening consequences. Several parameters need to be evaluated to ensure correct diagnosis. Pitfalls which can mask diagnosis using only a single test are renal failure, blood transfusion, diet or drugs. PMID- 9211195 TI - Inherited defects of purine and pyrimidine metabolism: laboratory methods for diagnosis. AB - The diagnosis of the majority of the known inherited defects of purine and pyrimidine metabolism can be achieved by analysing urinary excretion profiles. A quantitative measurement of the urinary uric acid/creatinine ratio should be the first approach for purine defects. The general screening system involves separation of the bases and nucleosides by reversed-phase high-performance liquid chromatography and multiwavelength UV detection. The catabolic defects of pyrimidine degradation can be diagnosed by gas chromatography-mass spectrometry as used for organic acids. For the detection of adenylosuccinase deficiency, several simple but effective thin-layer chromatographic methods are available. Techniques such as liquid chromatography-mass spectrometry, direct nega-tiveion fast-atom bombardment mass spectrometry, and proton nuclear magnetic resonance spectroscopy give promising results, but are not yet being used on a large scale. Patients should keep to a simple diet and preferably be free of medication in order to allow a reliable interpretation of the analytical data. PMID- 9211196 TI - Hepatic porphyrias in children. AB - Clinically overt hepatic porphyria is uncommon in children. The autosomal dominant acute hepatic porphyrias, acute intermittent porphyria (AIP), variegate porphyria (VP) and hereditary coproporphyria (HCP), are rarely present before puberty. Identification of asymptomatic children who have inherited these disorders is an important aspect of the management of the disease in their families and requires either enzymatic or DNA methods. Homozygous variants of AIP, VP and HCP usually present in early childhood and have phenotypes of variable severity. Mutational analysis is currently elucidating the relationship between these disorders and their autosomal dominant counterparts. 5 Aminolaevulinate dehydratase deficiency porphyria is a rare, autosomal recessive acute porphyria that may present at any age. Two cutaneous hepatic porphyrias are seen in children. Porphyria cutanea tarda (PCT), although mainly an adult disease, has been reported in young children with the autosomal dominant (type II) form of the disorder. Hepatoerythropoietic porphyria usually develops before the age of 2 years; patients are homo- or heteroallelic for uroporphyrinogen decarboxylase mutations, at least one of which is known to cause type II PCT. PMID- 9211197 TI - Gene transfer of the uroporphyrinogen III synthase cDNA into haematopoietic progenitor cells in view of a future gene therapy in congenital erythropoietic porphyria. AB - Congenital erythropoietic porphyria (CEP) is an inherited metabolic disorder characterized by an overproduction and accumulation of porphyrins in bone marrow. This autosomal recessive disease results from a deficiency of uroporphyrinogen III synthase (UROIIIS), the fourth enzyme of the haem biosynthetic pathway. It is phenotypically heterogeneous: patients with mild disease have cutaneous involvement, while more severely affected patients are transfusion dependent. The cloning of UROIIIS cDNA and genomic DNA has allowed the molecular characterization of the genetic defect in a number of families. To date, 22 different mutations have been characterized. Allogeneic bone marrow transplantation is the only curative treatment available for the severe, transfusion-dependent, cases. When bone marrow transplantation cannot be performed owing to the absence of a suitable donor, the autografting of genetically modified cells is an appealing alternative. The best approach to somatic gene therapy in this disease involves the use of recombinant retroviral vectors to transduce cells ex vivo, followed by autologous transplantation of the genetically modified cells. We investigated retroviral transfer in deficient human fibroblasts, immortalized lymphoblasts as well as bone marrow cells, and obtained a complete restoration of the enzymatic activity and full metabolic correction. Using K562 cells, an erythroleukaemic cell line, the expression of the transgene remained stable during 3 months and during erythroid differentiation of the cells. Finally, a 1.6- to 1.9-fold increase in enzyme activity compared to the endogenous level was found in normal CD34+ cells, a population of heterogeneous cells known to contain the progenitor/stem cells for long-term expression. The future availability of a mouse model of the disease will permit ex vivo gene therapy experiments on the entire animal. PMID- 9211198 TI - Erythropoietic protoporphyria. AB - Partial deficiency of the last enzyme of haem biosynthesis, ferrochelatase, leads to a distinct syndrome of photosensitivity caused by overproduction of protoporphyrin by erythropoietic tissue. Erythropoietic protoporphyria has an indeterminate pattern of inheritance and may be complicated by fulminating liver disease. The recent development of simple assays for ferrochelatase activity and cloning of the human ferrochelatase gene promises to shed light on the transmission of this disorder and may allow clinical expression of disease to be predicted. This review surveys the pathological features, genetics and treatment of porphyria. PMID- 9211199 TI - Disorders of homocysteine metabolism. AB - This review of recent advances covers (1) the metabolism of methionine and its regulation, emphasizing interactions with the three important vitamins folate, cobalamin and pyridoxine; (2) present knowledge of enzymological and moleculargenetic aspects of homozygous deficiencies of the three enzymes which cause elevated homocyst(e)ine; (3) recent clinical findings, post-methionine loading results related to enzyme and mutation studies in obligate heterozygotes for cystathionine beta-synthase deficiency; (4) important new evidence for disturbed homocysteine metabolism in neural tube defects, particularly based on studies of the thermolabile methylene-tetrahydrofolate reductase mutation which is also of importance in vascular disease; (5) the suitability and limitations of animal models that have so far been described. PMID- 9211200 TI - Assessment of homocysteine status. AB - Plasma total homocysteine (tHcy) determination is used in the diagnosis of homocystinuria, in cobalamin and folate deficiency and in cardiovascular risk assessment. However, determination of tHcy includes many pitfalls which complicate the assessment of homocysteine status. In the present article, we review basic knowledge for a rational use of plasma tHcy in diagnostic as well as scientific work. The subjects dealt with are procedures for sample handling and processing, the principles of tHcy analyses, and genetic and acquired determinants of the plasma tHcy concentration. PMID- 9211201 TI - The natural history of vascular disease in homocystinuria and the effects of treatment. AB - Among 40 patients with homocystinuria due to cystathionine beta-synthase deficiency diagnosed in the state of New South Wales, Australia (population 6 million) and followed long-term, there were 10 deaths at ages 2-30 years. Of these 8 were definite vascular deaths, one was a presumed vascular death, and the other was due to an accident and unrelated to homocystinuria. The vascular deaths were all early cases and only one patient, a pyridoxine-responsive 30-year-old woman, had been prescribed adequate treatment although it was uncertain that she was taking it. In 32 patients of mean age 30 years (range 9-66 years) there were 539 patient-years of treatment with pyridoxine, folic acid and hydroxocobalamin. There were 17 pyridoxine-responsive patients and all maintained plasma total free homocyst(e)ine levels < 20 mumol/L over an average treatment period of 16.6 years. The 15 nonresponsive patients received additionally 6-9 g of betaine daily. This resulted in a further 74% mean decline (+/-14% SD) in plasma total free homocyst(e)ine, persisting during an average (post-betaine) treatment period of 11 years; current mean +/- SD levels are 33 +/- 17 mumol/L (n = 15). There were two vascular events during treatment, one fatal pulmonary embolus (see above) and one myocardial infarction, whereas without treatment, 21 would have been expected, chi2 = 14.22, p = 0.0001, relative risk 0.09 (95% CI 0.02-0.38). There were no events during 258 patient-years of treatment in the 15 pyridoxine nonresponsive patients (p < 0.005 versus expected untreated). Nineteen patients had a total of 19 major and 15 minor operations requiring anaesthetic, and three had successful pregnancies, one whilst receiving betaine. There were no thromboembolic complications. We conclude that treatment which effectively lowers circulating homocyst(e)ine, even to suboptimal levels, markedly reduces cardiovascular risk in patients with cystathionine beta-synthase deficiency, and that betaine therapy contributes importantly to this in pyridoxine-nonresponsive patients. Betaine as additional therapy is safe and effective for at least 16 years. PMID- 9211202 TI - The case for mild hyperhomocysteinaemia as a risk factor. AB - The high incidence of vascular complications in severe hyperhomocysteinaemia in homozygotes for cystathionine beta-synthase deficiency has focused attention upon homocysteine as an atherogenic and thrombophilic agent. For two decades there has been accumulating evidence of mild hyperhomocysteinaemia as risk factor of vascular disease. Pooled data on hundreds of coronary, cerebrovascular and peripheral arterial disease patients show that mild hyperhomocysteinaemia was detectable in about 20-30%. In a recent meta-analysis of 27 studies up to 1994, including about 4000 patients and as many controls, it is calculated that the summary odds ratio of elevated homocysteine levels was 1.7, with 95% confidence interval (CI) 1.5-1.9, for coronary heart disease; it was 2.5, with 95% CI 2.0 3.0, for cerebro-vascular disease; and it was 6.8, with 95% CI 2.9-15.8, for peripheral vascular disease. The relevance of this newly recognized risk factor will be demonstrated by the outcome of the European Comac study on 'Hyperhomocysteinaemia and Vascular Disease', a multicentre case-control study on 800 vascular patients and 750 controls. Despite the selection for epidemiological reasons of a relatively low cut-off level as the criterion for mild hyperhomocysteinaemia in this study-the upper 20% of the distribution of control levels-the relative risk of thus-defined hyperhomocysteinaemia for arterial disease is about 2. This equals the relative risk of hypercholesterolaemia and of smoking; hypertension leads to a higher excess risk. The observed synergistic interaction between hyperhomocysteinaemia and hypertension and smoking may warrant a change in the now generally followed procedure of screening for hyperhomocysteinaemia only if conventional risk factors have not been detected in the patient. Those vascular patients with combined risk factors leading to synergism in their joint effect may profit most from homocysteinelowering intervention. PMID- 9211203 TI - Putative mechanisms for vascular damage by homocysteine. AB - In homocystinuria homocysteine appears to be directly toxic to the vasculature, but in mild hyperhomocysteinaemia a cause and effect relationship remains unproven. Evidence for a causal role is derived from recent primate and human studies in which endothelial dysfunction was produced by modestly elevated blood homocysteine concentrations. Endothelial dysfunction would account for an increased risk of both arterial and venous disease. A key abnormality may be impaired release and/or action of nitric oxide in response to flow. Other possible mechanisms include smooth muscle cell proliferation, extracellular matrix modification and lipoprotein oxidation. Although demonstrated in vitro, a role for lipoprotein oxidation in man has not been substantiated. However an effect of homocysteine on cellular redox status remains a possible mechanism. Homocysteine does not appear to alter circulating coagulation factors consistently, but may promote enhanced thrombin production indirectly by its effects on endothelium. Further studies are required to elucidate the pathological actions of homocysteine, concentrating on the effects of mild hyper homocysteinaemia on endothelial function in man. PMID- 9211205 TI - Quality in occupational health services. PMID- 9211204 TI - The role of vitamins in the pathogenesis and treatment of hyperhomocyst(e)inaemia. AB - The relation between vitamin nutritional status and circulating plasma homocyst(e)ine concentrations is reviewed. Several studies have shown that plasma concentrations of folate, vitamin B12 and pyridoxal 5'-phosphate are inversely associated with plasma total homocyst(e)ine concentrations. Of the three vitamins mentioned above, folate is the most powerful homocyst(e)ine lowering agent and a daily supplement of 0.65 mg/day is sufficient to normalize moderate hyperhomocyst(e)inaemia in most individuals with normal renal function. In patients with severe renal failure, high doses of folate are required to treat hyperhomocyst(e)inaemia. Folic acid is ineffective in reducing plasma total homocyst(e)ine concentrations in patients with a vitamin B12 deficiency. Vitamin B6 supplementation has no effect on fasting plasma total homocyst(e)ine concentrations, but attenuates the post-methionine load plasma homocyst(e)ine peak. At least one report has shown that some individuals appear to be unable to maintain plasma total homocyst(e)ine concentrations in the normal reference range by a dietary intake of folic acid only. Long-term vitamin supplementation may be indicated in these individuals. However, the clinical benefit of vitamin supplementation has not yet been demonstrated and controlled trials are urgently required. PMID- 9211206 TI - Quality in occupational health services. PMID- 9211207 TI - The American Board of Independent Medical Examiners. PMID- 9211208 TI - Assessment of exposure to polycyclic aromatic hydrocarbons during firefighting by measurement of urinary 1-hydroxypyrene. AB - Firefighters may be exposed to carcinogenic agents in the smoke from fires, and there has been some concern regarding firefighters' risk of developing occupational-related cancer. Polycyclic aromatic hydrocarbons (PAHs) are present in most fires, posing a cancer risk. The objective of this study was to evaluate the PAH exposure among firefighters. Students (n = 9) and teachers (n = 4) at a firefighter training school delivered urine samples before and 6 to 7 hours after extinguishing burning diesel fuel. The urine samples were analyzed by high performance liquid chromatography for 1-hydroxypyrene. A small but significant increase in 1-hydroxypyrene levels in the urine was found after the firefighting. This means that firefighting may cause exposure to PAHs. Although the exposure levels were low in this study, they may be different during other types of fires. PMID- 9211209 TI - Worksite characteristics and changes in worksite tobacco-control initiatives. Results from the COMMIT study. AB - Few studies have prospectively examined the characteristics associated with worksite adoption of tobacco-control initiatives. Data were collected as part of the Community Intervention Trial (COMMIT) for Smoking Cessation, which conducted interventions in 11 communities. This smoking cessation intervention was based on community organization principles and delivered through multiple community channels, including worksites, health care providers, the media, and cessation resources. This article reports results from telephone interviews of intervention community worksites having 50 or more employees, conducted at baseline and the end of the intervention period. Among worksites that responded to both baseline and final surveys, 83% had not adopted a smoke-free policy at baseline, and 61% did not offer any cessation aid or quitting resources at baseline. By the final survey, 34% of those with no smoking ban at baseline had become smoke-free, and 36% of those offering no cessation assistance at baseline were offering cessation resources at the follow-up. The prevalence of policy adoption was higher among worksites employing more female employees and offering other health-promotion activities; manufacturing businesses were significantly less likely than businesses other than service and wholesale/retail businesses to adopt policies. Adoption of cessation programs was significantly more likely among worksites employing 100 to 249 workers, compared with those employing 50 to 99 workers; those predominantly employing men; those offering other types of health-promotion activities; and those with a higher rate of turnover. These results provide important information about the characteristics of worksites likely to engage in tobacco-control efforts. Health educators and others may choose to target those worksites most ready for adoption of tobacco control policies and programs, as indicated by these findings. PMID- 9211210 TI - Elevated serum liver enzymes in coke oven and by-product workers. AB - Coke oven and by-product workers are potentially exposed to coke oven emissions (COE), which contain hundreds of chemicals and are primarily composed of polycyclic aromatic hydrocarbons (PAH) and volatile organic compounds. Some of these compounds are hepatotoxins. The objective of this study was to examine the relationship between work in coke oven and by-product plants and serum activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), the most commonly performed liver-function tests. The exposed group was composed of current workers who had been employed at least 3 months in the two coke-operation work areas, including one coke oven plant and one by-product plant (Area I: n = 117; Area II: n = 96) of a large steel company in Taiwan. Control subjects (Area III: n = 131), not visiting either coke-operation area in the last 3 months, were collected from the administrative and nonproduction areas in the same company. PAH exposure, as a surrogate of COE, was measured monthly by PM-10 size-selective high-volume-area air samplers in or around these three areas between June and December 1990, as well as between November 1992 and June 1993. The mean total respiratory particulate PAH exposure levels (< 10 microns) between November 1992 and June 1993 in Area I, II, and III were 6.8 x 10(3), 2.1 x 10(3), and 6.5 x 10(1) ng/m3, respectively. AST, ALT, and hepatitis B surface antigen tests were performed in 1994. Workers who showed either AST or ALT levels greater than reference levels (abnormal > 25 IU/L) were regarded as showing "elevated liver enzyme levels." Workers in Area I had AST levels that were 17% higher (95% confidence interval [CI], 3% to 32%]) and ALT levels that were 35% higher (95% CI, 10% to 65%)] than those in Area III after controlling for appropriate confounders. The adjusted odds ratio (Area I vs Area III) for elevated liver enzymes was 4.4 (95% CI, 1.5 to 13.4). In addition, coke oven (n = 91) and by product workers (n = 26) from Area I had ALT levels 37% and 45% higher, respectively, compared with control subjects from Area III, after adjusting for appropriate confounders. Similar effects are also seen for AST. Workers in Area II had slightly, but not significantly, elevated AST and ALT levels. These results indicate that workers most heavily exposed to COE exhibit elevated aminotransferase levels. PMID- 9211211 TI - Occurrence of asthma and chronic bronchitis among female hairdressers. A questionnaire study. AB - We carried out a retrospective cohort study using a self-administered questionnaire to assess the risk of hairdressers to develop asthma and chronic bronchitis. A representative sample of 4433 female hairdressers and an equal number of shop personnel in employment in 1980 was drawn from the Longitudinal Census Data File of Statistics Finland. Physician-diagnosed asthma, chronic bronchitis, and other respiratory diseases in 1980 and 1995 were inquired about in the respiratory part of the questionnaire. The response rate to the questionnaire was 82% for the hairdressers (n = 3484) and 79% for the referents (n = 3357). The prevalence of asthma among the hairdressers was 5.6% in 1980 and 10.1% in 1995, and the prevalence of chronic bronchitis was 3.9% in 1980 and 5.6% in 1995. The relative risk for asthma (odds ratio [OR]: 1.7, 95% confidence interval [CI]: 1.3 to 2.3 in 1980; and OR: 1.7, 95% CI: 1.4 to 2.2 in 1995) and for chronic bronchitis (OR: 2.2, 95% CI: 1.5 to 3.2 in 1980; and OR: 1.9, 95% CI: 1.4 to 2.6 in 1995) among hairdressers was almost twice that in the reference group. The incidence rate of asthma in 1980 through 1995 was 2.2 and of chronic bronchitis was 1.1 cases per 1000 person-years among hairdressers, whereas the rate in the reference group was 1.3 asthma cases and 0.9 chronic bronchitis cases per 1000 person-years. The relative risk for developing asthma during the 15 years observation time was 1.7 (95% CI: 1.1 to 2.5) and for chronic bronchitis was 1.2 (95% CI: 0.7 to 1.9) among hairdressers, compared with referents. Our results indicate that hairdressers are at higher risk for developing asthma. PMID- 9211212 TI - Comparison of in vivo and in vitro measures of beryllium sensitization. AB - Chronic beryllium disease (CBD) diagnosis hinges on demonstrating a cell-mediated immune response to beryllium salts in vitro with the beryllium lymphocyte proliferation test (BeLPT). The BeLPT has found widespread application in screening for CBD and beryllium sensitization in populations of exposed workers. We hypothesized that the in vivo beryllium salt patch test may be of value as an adjunct to the BeLPT, rectifying false negative or ambiguous blood test results. We studied subjects with CBD (n = 11), beryllium sensitization without disease (n = 3), and control subjects with dermatitis (n = 20). Evaluation included completion of a demographic questionnaire, blood BeLPT (if CBD or beryllium sensitized), and beryllium patch testing with 0.1% and 1% beryllium sulfate (BeSO4) in petrolatum and in aqueous vehicles. Biopsies were performed at abnormal patch test sites in five subjects. The 1% aqueous BeSO4 proved superior either to 1% petrolatum or 0.1% solutions, producing positive reactions in all CBD and beryllium-sensitized subjects. We observed no long-term adverse reactions. Biopsies demonstrated spongiotic changes early, followed by noncaseating granulomas within 18 days. We conclude that the beryllium patch test can be used safely to clarify the sensitization state and diagnosis of CBD. PMID- 9211213 TI - Historical cohort mortality study of a continuous filament fiberglass manufacturing plant. II. Women and minorities. AB - An historical cohort mortality study was undertaken at Owens Corning's continuous filament fiberglass manufacturing plant in Anderson, South Carolina. The cohort included 1074 white women, 130 black women, and 494 black men who worked for a minimum of one year from the opening of the plant in 1951 through December 31, 1991. This represents the largest single cohort of white women assembled to date in either a wool or continuous filament fiberglass manufacturing facility and represents the first study of a cohort of black men and women in the man-made vitreous fiber industry. Over 95% of the women and minorities included in this report held production positions in the plant. There were no significant excesses or deficits in mortality by cause, including cancer causes, among white women, with the exception of motor-vehicle accidents, when compared with national mortality. Among black men, standardized mortality ratios (SMRs) for heart disease are significantly below one, and SMRs for all cancers combined are below unity on both national and local standards. Lung cancer SMRs are below unity for both white women and black men. PMID- 9211214 TI - Self-reported stress and reproductive health of female lawyers. AB - We studied the prevalence and relationship of stress and working conditions with adverse reproductive outcomes in a cohort of female US law-school alumnae. A total of 584 female lawyers (74% response), aged 25 to 63, responded to a mailed questionnaire. Job hours per week was a strong predictor of job stress. In a logistic regression analysis, women working > 45 hours/week were five times as likely to report high stress as those working < 35 hours/week. Marriage and length of time on the job showed a small inverse association with stress. Women who worked more than 45 hours/week during their first trimester of pregnancy were more likely to report high stress at work during pregnancy. After being adjusted for confounding factors, weekly job hours during the first trimester of pregnancy showed a strong independent association with spontaneous abortion risk (odds ratio [OR], 3.0; 95% confidence interval [CI], 1.4 to 6.6). Seven or more alcohol drinks/week was also independently associated with spontaneous abortion risk (OR, 4.8; 95% CI, 1.5 to 18.1). Self-reported stress during pregnancy was positively but not statistically significantly associated with spontaneous abortion (OR, 1.4; 95% CI 0.8 to 2.3). PMID- 9211215 TI - A topography of psychiatric disorders among correction officers. AB - This study assessed the nature, scope, and work-related impact of psychiatric disorders among correction officers. It also examined the effect of psychiatric diagnoses on occupational functioning. Medical charts of 1029 correction officers with complaints of psychological distress were reviewed for DSM-III-R diagnoses and duration of disability. Subjects evinced a wide range of psychiatric conditions, accounting for a substantial loss of full-duty workdays. The most frequently observed diagnoses were V Codes, mood and adjustment disorders. The most disabling conditions in terms of duration of disability were mood, anxiety, and personality disorders. V Codes accounted for the highest cumulative duration of disability among all officers. Results suggest that psychiatric disability among correction officers is most often a function of V-Code conditions, as opposed to major Axis I or II psychopathology. PMID- 9211216 TI - Occupational exposure to antineoplastic agents and self-reported infertility among nurses and pharmacists. AB - Although infertility has been identified as an effect of chemotherapy for some cancer patients, the association of infertility with occupational exposure has not been investigated. This case-control study investigated the relationship of infertility with occupational handling of chemotherapy drugs by nurses and pharmacy personnel. Data were gathered by questionnaire from 4659 staff at facilities participating in the National Surgical Adjuvant Breast and Bowel Project collaborative clinical trials network of the National Cancer Institute. The 405 subjects reporting infertility were each matched by sex and age with three control subjects and compared for history of chemotherapeutic drug handling. Results for the total sample and for women showed a significantly elevated odds ratio (OR = 1.5; CI = 1.1 to 2.0) for self-reported infertility associated with occupational handling of chemotherapeutic drugs prior to onset of infertility. For men, the odds ratio was similar but not statistically significant. This worker population, with a mean age of 37, is in the prime of reproductive life. Prevention of chemotherapy side effects by use of available protection is preferable to risking infertility. PMID- 9211217 TI - Crystal structure of a dipeptide Boc-Aib-Phe-OMe. AB - In order to understand the effect of the restrictions posed by the Aib residue on peptide conformation we studied the crystal structure of a dipeptide tBoc-Aib-Phe OMe. Crystals of this compound are triclinic, space group P1 with a = 9.600(1) A, b = 10.262(1) A, c = 10.799(1) A, alpha = 98.43 degrees (1), beta = 99.18 degrees (1), gamma = 98.87 degrees (1), V = 1021.69(18) A3 and Z = 2. The structure was solved by direct methods and refined to an R-factor of 4.98%. The backbone conformational angles for the Aib residue in molecule A are in the left-handed helical region, while in molecule B they are in the right-handed helical region. The Phe residue in molecule A is in the right-handed helical conformation, while in molecule B it is in the beta-region. The peptide units are trans and show significant deviation from planarity [(omega 1 = 166.67(5) degrees and omega 2 = 177.9(5)]. PMID- 9211218 TI - Comparison of the photochemical behavior of four different photoactivatable probes. AB - The most commonly used photoaffinity labeling probes are compared, which are aryl azides, aryl diazirines, alpha-diazocarbonyls and benzophenone-derivatives. The compounds were used under identical conditions and crosslinking efficiency, influence of water, irradiation requirements, and by-products were investigated. Using the pentapeptide thymopentin (TP5) as a model system, we synthesized four analogues by solid-phase peptide synthesis and partially N-terminal modification to obtain [p-(3-trifluoromethyl)diazirinophenylalanine5]TP5, [p benzoylphenylalanine5]TP5, 4-azidobenzoyl-TP5 and 2-diazo-3,3,3 trifluoropropionyl-TP5. The peptides were characterized by HPLC and ion-spray mass spectroscopy. Irradiation of the peptides with two different ultraviolet sources was carried out in water, n-propanol and water/n-propanol to imitate both hydrophobic and hydrophilic peptide/protein-interactions as well as the influence of the aqueous environment. Analysis of the products with HPLC, ion-spray MS, HPLC-MS and HPLC-CID-MS revealed that (Tmd)Phe is a highly potent carbene precursor, which can be transformed easily into uniform crosslinking products by smooth photolysis. However, the electrophilic nature of the intermediate causes a high tendency to react with water molecules. The 4-azidobenzoyl group showed comparable crosslinking efficiency, but the probability to create non-uniform irradiation products (e.g. through rearrangement) is higher, whereas the reaction with water is less dominant. In contrast, Bpa was found to have an extremely low affinity to react with water, whereas prolonged UV irradiation is needed to get complete rearrangement into a variety of products. As the absorption band of alpha-diazocarbonyls at around 350 nm possesses a low extinction coefficient, 2 diazo-3,3,3-trifluoropropionyl-TP5 could not be activated at all with the optimized irradiation conditions that we have chosen for our comparative studies. PMID- 9211219 TI - X-ray structure of Tyr-D-Tic-Phe-Phe-NH2 (D-TIPP-NH2), a highly potent mu receptor selective opioid agonist. Comparison with proposed model structures. AB - Tyr-D-Tic-Phe-Phe-NH2 (D-TIPP), a linear tetrapeptide containing the conformationally restricted Tic residue (tetrahydroisoquinoline-3-carboxylic acid), is an opioid agonist which exhibits high affinity and selectivity for the mu-receptor. Its conformational features have been studied using a combination, a solid-state (X-ray) and modeling (molecular mechanics and Monte Carlo simulations) methods. The results of the X-ray study showed two distinct conformers for D-TIPP, with the main differences lying in the orientation of the Tyr side-chain and the presence of both D-Tic(+) and D-Tic(-) conformations for the D-Tic residue. The peptide backbone is folded and stabilized by the formation of one intramolecular hydrogen bond. The modeling results also indicated a folded backbone for the peptide and both cis and trans conformers for the D-Tic residue are found in the lowest-energy structures. Comparison of the X-ray and modeling results shows many similarities especially around the D-Tic residue. PMID- 9211220 TI - Conformational stability of a type II' beta-turn motif in human growth hormone [6 13] peptide analogues at hydrophobic surfaces. AB - The interactive properties of several peptides related to human growth hormone (hGH) [6-13] containing a type II' beta-turn motif have been investigated using reversed-phase high-performance liquid chromatography (RP-HPLC). Various chromatographic parameters related to the hydrophobic interactive surface area and binding affinity were measured over the range of temperatures between 5 and 85 degrees C. Variations in these parameters were consistent with significant differences in the relative stability of the type II' beta-turn structures of these peptidomimetics. The effect of changes in peptide conformation were also investigated through the analysis of band-broadening behaviour during the chromatographic process. Significant variations in bandwidth observed at discrete temperatures were related to the rate of interconversion between the type II' beta-turn and more extended conformers. These investigations further document the potential of RP-HPLC for monitoring subtle changes in peptide secondary structure at hydrophobic interfaces. PMID- 9211221 TI - Conformational consequences of i, i + 3 cystine linkages: nucleation for alpha helicity? AB - Methods to introduce specific secondary structural elements into peptides and proteins are vital for the rational design of peptide and non-peptide drug candidates as well as in the de novo design of proteins. Here the incorporation of a disulfide linkage between cysteine residues spaced three amino acids apart (i, i + 3) as a method to induce helicity is examined. Two dodecamer peptides, A V-S-E-C-Q-L-C-H-D-K-G-NH2, differing in the chirality of the cysteine at the fifth position (the i position), have been synthesized and conformationally studied both in the linear and cyclized form. This peptide sequence, derived from the N-terminal sequence of parathyroid hormone related protein, does not form helices, even as part of the 1-34 fully active domain of the protein. The four analogs (two cyclic and two linear) were analyzed both in aqueous solution and in the presence of sodium dodecyl sulfate micelles. In aqueous solution the linear peptides display no evidence for secondary structure, while the cyclization induces a turn centered about the cysteine residues. In the presence of micelles the linear form of the peptides adopts bent conformations, containing turns, but results from both NMR and CD provide no evidence of helices. The oxidized L,L peptide in the micellar solution does not present a well defined conformation, although the presence of one helical turn is evident. The cyclic D,L analog adopts a helical structure (not an alpha-helix) extending from residue 2 to 9, with non-standard phi, psi values caused by the presence of the D-amino acid. These results clearly illustrate that the ability of D-Cys(i), Cys(i + 3) cyclization to initiate helix formation depends greatly on the solvent used. Therefore, any drug-design principle utilizing this modification for helix nucleation must keep the environment in which the peptide is biologically active in mind. PMID- 9211222 TI - Sulfonated analogues of cyclolinopeptide A. Synthesis, immunosuppressive activity and CD studies. AB - Linear and cyclic analogues of cyclolinopeptide A (CLA) in which one or both phenylalanine residues in fragment Pro6-Pro7-Phe8-Phe9 were substituted by their sulfonated derivatives have been synthesized by SPPS method and cyclization with the BOP reagent. The peptides were examined for their immunosuppressive activity in the humoral and cellular immune response by PFC and DTH tests. All of the analogues retain some immunosuppressive activity of native CLA. Their CD spectra confirm that the optical activity of aromatic residues in CLA depends on their position in the peptide chain. Only the residue in position 8 seems to be optically active. CD spectrum of the cyclic analogue modified in position 9 is very similar to that of native CLA which correlates with its high biological activity. The chiroptical properties of the p-sulfonated Phe-residue are established. PMID- 9211223 TI - Synthesis and biological activity of novel backbone-bicyclic substance-P analogs containing lactam and disulfide bridges. AB - A biased library of 60 novel backbone-bicyclic Substance P analogs was prepared by the simultaneous multiple peptide synthesis method. The peptides, containing both a lactam and a disulfide ring, were synthesized by combined Boc and Fmoc chemistries, and were cyclized on the resin. Cleavage of the S-benzyl group and oxidation of the sulfhydryl groups was enabled by adaptation of the diphenylsulfoxidetrichloromethylsilane method to solid-phase synthesis. The peptides were screened for NK-1 and NK-3 activity, and were found to be weak agonists. PMID- 9211224 TI - Role of tryptophan-14 in the interaction of dynorphin A(1-17) with micelles. AB - Fluorescence spectroscopy has been used to examine the interaction between the opioid peptide dynorphin A(1-17) (dynorphin) and dodecylphosphocholine (DPC) micelles. Fluorescence emission spectra as a function of added lipid indicate insertion of the Trp14 side chain into the hydrophobic portion of the micelle, supporting NMR results from this laboratory. A model of interaction with micelles consistent with the fluorescence results and earlier NMR results is proposed. The critical micelle concentration in the presence of peptide was also determined, and is discussed in the context of relevance to both NMR spectroscopy and peptide lipid interactions. PMID- 9211225 TI - NMR and circular dichroism studies on the conformation of a 44-mer peptide from a CD4-binding domain of human immunodeficiency virus envelope glycoprotein. AB - Two-dimensional NMR, circular dichroism (CD) experiments and molecular modeling were performed to study the secondary structure of a 44-mer peptide fragment derived from the C4 region of gp120 of human immunodeficiency virus in aqueous solution. It was found a nascent helical structure exists following a type I turn near the N-terminus of the peptide. The proline residue in the turn appears to serve as a helix initiator. The helical structure was in fast dynamic equilibrium with beta- or random coil form on the NMR scale. A reverse turn was identified at a section containing two consecutive proline residues. A nascent helical structure has been detected for the region near the C-terminus of the 44-mer peptide. Higher helical content for the peptide is also indicated by CD studies on TFE titration. Thus it is proposed that, in more apolar medium, the Pro-Pro turn and the segment amino-terminal to it, spanning about 20 amino acids, may be converted into helix structure. Moreover, the region near the C-terminus of the peptide may also be induced into helix, so that a helix-turn-helix structure may be formed in the C4 domain of gp120. A helical wheel representation of this stretch shows amphipathicity of the helix. The biological implication of the conformational adaptibility of the peptide was discussed. PMID- 9211226 TI - Conformational dynamics and kinetics of peptide antagonist interactions with interleukin-1 receptor. Fluorescence studies using the NBD-labelled peptide AF12415. AB - Interleukin-1 plays a key role in the inflammatory response provoked by various disease states and inhibition of its action can bring therapeutic benefits. Steady-state and time-resolved studies of the intrinsic tryptophan fluorescence of the free soluble Type I form of interleukin-1 receptor (IL-1R) reveal that the rotational motions of the three major domains are strongly associated. Bound peptide antagonists are buried in hydrophobic regions, but a flexible association permits access to species from the aqueous phase. Ligand binding does not lead to rigidification of the receptor structure. The kinetics and mechanism of complex formation and dissociation, involving IL-1R with receptor antagonist protein (IL 1ra) and with peptides AF11733 (15 aa) and AF10961 (21 aa) were determined with the aid of peptide AF12415 (15 aa) labeled at its N-terminus by the NBD fluorophore, which exhibits a five-fold increase in emission intensity at 540 nm on binding of the peptide to IL-1R. The magnitude of the ON rate constant, typically 1 x 10(6) M-1 s-1, implies the existence of an intermediate 'encounter complex' involving interactions of low specificity. Readjustments of the initial encounter complex leads to a final complex where very specific interactions dominate. The first-order rate constant for this latter process is the most sensitive indicator of the true peptide affinity for the receptor binding site, and thus provides a better criterion than the apparent OFF rate (typically 2 x 10(-3) s-1) for discrimination of peptide antagonists. PMID- 9211227 TI - Emerging diseases--what veterinarians need to know. AB - The recent increase in emerging diseases can be attributed to a number of factors, all of which relate to some form of alteration in the way etiologic agents move around. Some of these factors responsible for altered agent trafficking include actual transport to a susceptible population or new species, environmental disruption that facilitates exchange of microbes, and a husbandry change that promotes new ways for microbes to move around. Given the exponential growth of the human population and all the attendant implications, including the mobility of this population, the ecological disruption that is accompanying the overall increase, and the necessity of exploring new agricultural technologies to feed a burgeoning population, it is a certainty that altered agent trafficking will not only continue but will undoubtedly increase. Veterinarians should be aware of the role they will be expected to play in this field. PMID- 9211228 TI - Bluetongue virus detection: a safer reverse-transcriptase polymerase chain reaction for prediction of viremia in sheep. AB - A reversible target capture viral RNA extraction procedure was combined with a reverse-transcriptase nested polymerase chain reaction (PCR) to develop a capture PCR assay providing a rapid and safe prediction method for circulating bluetongue virus in infected ruminants. This new assay was compared with virus isolation and a recently developed antigen-capture enzyme-linked immunosorbent assay (ELISA) for the detection of bluetongue virus. Eight Warhill crossbred sheep were inoculated subcutaneously with bluetongue virus serotype 10, and blood samples were taken sequentially over a period of 28 days. The capture PCR detected the peak of viremia, as determined by virus isolation and antigen-capture ELISA, from day 5 to day 14 after challenge. The results indicate that the rapid-capture bluetongue virus PCR provides a rapid indicator of samples in which virus can be isolated. In addition, this capture bluetongue virus PCR procedure does not require a lengthy phenol extraction or the use of the highly toxic methyl mercury hydroxide denaturant. PMID- 9211229 TI - Serologic reactivity using conserved envelope epitopes in feline lentivirus infected felids. AB - An enzyme-linked immunosorbent assay (ELISA) based on synthetic peptides identical to lentivirus envelope protein amino acid sequences was used to study serologic reactivity of lentivirus-infected domestic cats and nondomestic felids. One feline immunodeficiency virus (FIV) peptide, P237, was consistently recognized by antibodies from FIV-infected cats, but 2 other FIV peptide antigens were not. The molecular basis for this serologic reactivity was examined. Lentivirus-infected nondomestic Felis species reacted intensely with a puma lentivirus (PLV) peptide corresponding to the conserved FIV peptide. However, lentivirus-infected Panthera species, from which a different lentivirus has been isolated, did not react with the PLV. FIV-infected domestic felids also did not have significant reactivity with the PLV peptide. The peptide ELISA is comparable in sensitivity and specificity to western blot analysis and a commercial enzyme immunoassay. Unlike the other assays, however, the peptide ELISA is inexpensive, requires a small amount of serum, enables the study of specific isotype reactivity, and discriminates between antibodies to FIV and those to PLV. Antibody tests based upon the FIV and the PLV peptides should be useful for detecting the possible introduction of FIV into exotic felids or of lentiviruses from nondomestic felids into the domestic cat population. PMID- 9211230 TI - Development and evaluation of a recombinant antigen, monoclonal antibody-based competitive ELISA for heartwater serodiagnosis. AB - Cowdria ruminantium is the etiologic agent of heartwater, a tick-transmitted foreign animal disease with considerable potential for entrance into the USA. A competitive enzyme-linked immunosorbent assay (cELISA) was developed to detect serologic responses to C. ruminantium infection. The cELISA utilized a recombinant form of the C. ruminantium major antigenic protein (MAP-1) as the antigen and an anti-MAP-1 monoclonal antibody as the competing indicator reagent. Experimental antisera to C. ruminantium and a wide variety of related ehrlichial organisms were used to evaluate cELISA reactivity. Only sera against C. ruminantium, Ehrlichia canis, E. chaffeensis, and a recently discovered cervine ehrlichia-like organism reacted positively in the cELISA. Specificity of the cELISA was > or = 99.5% in a survey of 1,774 southeastern US and Puerto Rican slaughter cattle sera but was only 85% in a group of 79 hunter-killed white tailed deer (Odocoileus virginianus) from the southeastern USA. Reference true positive and cELISA false-positive sera were further analyzed by end point titrations using the cELISA and by indirect fluorescent antibody (IFA) tests for reactivity with C. ruminantium, E. canis, and E. chaffeensis antigens. True heartwater-positive sera were significantly more reactive using the cELISA and C. ruminantium IFA procedures (P < 0.05), whereas false-positive sera were significantly more reactive with the antigens used in the E. chaffeensis IFA procedure (P < 0.05). A group of sera from 210 field-origin ruminants residing on known or potentially heartwater-endemic Caribbean islands revealed a substantial (12.4%) prevalence of cELISA-positive specimens. The cELISA is a relatively specific serodiagnostic test for heartwater in cattle and could be used to monitor for possible introduction of the disease into the USA. The cELISA may also be an excellent tool for monitoring the success of an ongoing Caribbean Amblyomma tick eradication program designed to eliminate the biological vector responsible for the perpetuation and spread of this dangerous foreign animal disease. PMID- 9211231 TI - Experimental vesicular stomatitis in swine: effects of route of inoculation and steroid treatment. AB - An enzootic focus of vesicular stomatitis virus New Jersey serotype (VSV-NJ) exists on Ossabaw Island, Georgia. Many questions regarding the epizootiology of this virus at this focus still exist, but evidence suggests that the vector for this virus is a phlebotomine sand fly (Lutzomyia shannoni), with feral swine serving as a potential source of virus for the sand fly and for other swine via contact transmission. We conducted 2 experimental trials in domestic swine using VSV-NJ isolated from a sand fly from Ossabaw Island to determine if route of inoculation or immunosuppression via steroid administration affected the development of disease, viremia, viral shedding, or the neutralizing antibody response. In a third trial, we studied the potential for contact transmission among swine using this isolate. Virus isolations were made from nasal cavity or palatine tonsil of the soft palate, and VSV-NJ neutralizing antibodies developed when pigs were inoculated intradermally in the apex of the snout, ear, or coronary band, intravenously, intranasally, or via scarification of the apex of the snout or coronary band. Vesicles developed only in pigs inoculated in the apex of the snout or coronary band, and these vesicles were at the site of inoculation. Steroid treatment did not potentiate the development of secondary vesicles and did not prolong the period of virus shedding from VSV-NJ-infected swine. Contact transmission, as determined by shedding of virus from the tonsil of the soft palate and the development of VSV-NJ neutralizing antibodies, occurred in pigs in contact with animals inoculated in the apex of the snout but not in contact animals exposed to pigs inoculated intradermally in the coronary band or intranasally. These trials show that contact transmission can occur and VSV-NJ can be shed without the development of clinical disease (i.e., vesicle formation). Viremia was never detected in any of the experimental pigs, suggesting that swine may not be a good amplifying host for VSV-NJ. PMID- 9211232 TI - Restriction endonuclease analysis of equine herpesvirus-1 isolates recovered in Ontario, 1986-1992, from aborted, stillborn, and neonatal foals. AB - Ninety-two equine herpesvirus type 1 isolates were recovered from aborted, stillborn, or neonatal foals from Ontario, Canada, from 1986 to 1992. From this total, 32 strains were randomly chosen for further study. Four or 5 isolates from each winter were selected, each from a different premises, and characterized by restriction enzyme analysis using BamHI, KpnI, BglII, HindIII, and EcoRI. Additional isolates from 2 premises and from a zebra foal were also assessed. For the strains isolated in 1986 and 1989-1992, the DNA pattern of 18 strains was similar to that of type 1P (Kentucky D) for BamHI and KpnI. None of the 32 strains studied could be differentiated by HindIII or EcoRI. Using BglII, an inconsistent fragment pattern and distribution were observed. Of the 8 strains isolated in 1987 and 1988, 7 were assigned into the 1B prototype group. The geographic distribution of 17 type 1P and 12 1B isolates was random across southern Ontario. These findings suggest that both electropherotypes can be recovered from horses in Ontario. The patterns of the additional equine isolates from the same premises were identical. The zebra isolate was different from the prototype equine herpesvirus type 1 and type 4 patterns and from all other equine isolates. PMID- 9211233 TI - A novel protein polymorphism differentiates the California serotype of infectious bronchitis from other serotypes common to California. AB - The California (Cal) serotype of infectious bronchitis virus (IBV) was isolated from layer flocks in southern California in the early 1980s. Since then, it has spread to the broiler-producing regions of central California, where it has been implicated in respiratory disease outbreaks in vaccinated flocks. Lack of a procedure for quickly identifying IBV serotypes in commercial chicken flocks has prevented the causal association of the IBV Cal serotype with respiratory disease outbreaks. A protein polymorphism has been identified in the matrix protein of the Cal serotype; it appears to be unique among other common serotypes of infectious bronchitis virus found in California. This polymorphism can be identified on western blots using raw or concentrated infectious allantoic fluid as the source material. Identification of the Cal serotype and of serotypes in the Mass and Conn groups can be performed rapidly using field samples from suspect flocks. The identification of this polymorphism provides an alternative method for the rapid identification of the Cal serotype of IBV. PMID- 9211234 TI - An improved method for the recovery of mycoplasmas from the external ear canal of goats. AB - The efficacy of ear canal flushing and ear canal and mouth swabbing methods for the isolation of mycoplasmas was investigated in 39 goats. Of the 19 goats positive for Mycoplasma spp., 14 (73.7%) were positive with the ear canal flushing method, 4 (21.0%) were positive with both ear canal flushing and mouth swabbing methods, and 1 (5.3%) was positive by the mouth swabbing method. Mycoplasma arginini, M. mycoides subsp. mycoides, and M. mycoides subsp. capri were identified by direct immunofluorescence and growth inhibition tests. Previous reports on the isolation of M. arginini from the ear canal of goats were not found in the literature. PMID- 9211235 TI - Two new serovars of Chlamydia psittaci from North American birds. AB - Five Chlamydia psittaci isolates (1 turkey, 1 psittacine, 1 human, and 2 pigeon isolates) failed to react with serovar-specific monoclonal antibodies to known avian and mammalian C. psittaci serovars and were presumed to represent 1 or more new serovars. The isolates were characterized using restriction endonuclease analysis of the whole genome, polymerase chain reaction-restriction fragment length polymorphism of the major outer membrane protein genome, monoclonal antibody comparisons, and growth in tissue culture. Monoclonal antibodies were produced to the human isolate (MP) and to the psittacine isolate (VS225). The monoclonal antibody results show that the isolates represent 2 new avian serovars (serovars E and F). The restriction fragment length polymorphism analysis of the major outer membrane protein genome demonstrated that the isolates are distinct. The whole genome restriction endonuclease analysis data and the growth patterns in tissue culture indicate that the new serovars are similar to avian serovars recognized previously. A subspecies monoclonal antibody that reacted with serovars A and B also reacted with serovar E, indicating that these serovars are closely related. The results show that these isolates represent 2 new avian serovars, making them the fifth and sixth avian serovars identified in North American birds. PMID- 9211236 TI - Laboratory identification and enteropathogenicity testing of Serpulina pilosicoli associated with porcine colonic spirochetosis. AB - Pathogenic intestinal spirochetes of swine include Serpulina hyodysenteriae, a strongly beta-hemolytic spirochete that causes swine dysentery, and S. pilosicoli, a weakly beta-hemolytic intestinal spirochete (WBHIS) that causes porcine colonic spirochetosis. Because of the existence of nonpathogenic WBHIS in the normal swine colon, it is important to develop laboratory procedures for accurate identification of S. pilosicoli. The purpose of the present study was to assess hippurate hydrolysis and polymerase chain reaction (PCR) amplification of specific 16S ribosomal RNA (rRNA) sequences for identification of porcine S. pilosicoli. Additionally, the enteropathogenicity of 8 field isolates of porcine S. pilosicoli was determined by challenge exposure of 1-day-old chicks and sequential histologic examination of the cecal mucosa. The field isolates of porcine S. pilosicoli hydrolyzed hippurate and yielded S. pilosicoli-specific products by PCR amplification of 16S rRNA sequences. Although all of the field isolates of porcine S. pilosicoli attached to the cecal epithelium of challenge exposed chicks by day 21 postinoculation, some isolates had locally invasive phenotypes. We concluded that identification of porcine S. pilosicoli could be made on the basis of results of hippurate hydrolysis and 16S rRNA PCR amplification. Challenge inoculation of 1-day-old chicks followed by histologic examination of the cecal mucosa demonstrated the enteropathogenicity of porcine S. pilosicoli. PMID- 9211237 TI - Investigation of the selenium status of aborted calves with cardiac failure and myocardial necrosis. AB - Lesions of heart failure, specifically cardiac dilation or hypertrophy along with a nodular liver (chronic passive congestion) and ascites, have been found in 4-5% of aborted bovine fetuses. In this study, a group of 22 such fetuses was compared with groups of aborted fetuses without lesions of heart failure and with nonaborted fetuses obtained from a slaughterhouse. The fetuses were necropsied, tissues were taken for histopathology, and samples were collected for routine bacteriologic and virologic examinations. Liver and kidney tissue was saved for selenium analysis. Histopathologic examinations of myocardium of fetuses with cardiac failure revealed myocardial necrosis and mineralization in 7 fetuses, lymphocytic myocarditis in 5 fetuses, myocardial fibrosis in 5 fetuses, or no microscopic lesions in 5 fetuses. Mean liver selenium levels were 5.5 mumol/kg in the fetuses with heart lesions, 6.5 mumol/kg in the fetuses without heart lesions and 7.5 mumol/kg in fetuses from the slaughterhouse; these differences were statistically significant. The results suggest that selenium deficiency in bovine fetuses may cause myocardial necrosis and heart failure. This study also provides data on normal liver and kidney selenium levels in bovine fetuses from the analyses of 19 nonaborted fetuses. PMID- 9211238 TI - Bovine fetal neosporosis: a comparison of epizootic and sporadic abortion cases and different age classes with regard to lesion severity and immunohistochemical identification of organisms in brain, heart, and liver. AB - Eighty bovine fetuses with confirmed neosporosis were used to score lesion severity and presence of parasites in brain, heart, and liver. A comparison was made between epizootic and sporadic abortion cases. The possible influence of fetal age was also investigated. Histologic lesions of multifocal encephalitis, myocarditis, and periportal hepatitis with or without focal hepatocellular necrosis were almost always observed. Neospora caninum tachyzoites were identified immunohistochemically in 85% of the brains, 14% of the hearts, and 26% of the livers. Tissue cysts were observed in 21% of the brains. Significant differences between epizootic and sporadic abortion cases were found only in the liver. Hepatic lesions were more prominent and N. caninum tachyzoites were observed more frequently and in higher numbers in epizootic cases. Examination by immunohistochemistry of the liver in addition to the brain can be highly contributive diagnostically, particularly in epizootic cases. There were no significant age-related differences except for a higher presence of tachyzoites in the hearts of younger fetuses (3-4 months gestational age). PMID- 9211239 TI - Polymerase chain reaction and restriction endonuclease digestion for selected members of the "Mycoplasma mycoides cluster" and Mycoplasma putrefaciens. AB - A specific diagnostic method using the polymerase chain reaction, together with restriction endonuclease digestion patterns, was developed for members of the "Mycoplasma mycoides cluster" that normally occur in the United States (i.e., Mycoplasma mycoides subsp. mycoides Large Colony and Mycoplasma capricolum subsp. capricolum in addition to "cluster" mycoplasma, bovine serogroup 7, and Mycoplasma putrefaciens. The digestion of "cluster" polymerase chain reaction DNA (1,225 bp) amplification products with restriction enzymes AseI and SspI gave mycoplasma species-specific patterns for all strains of M. mycoides subsp. mycoides Large Colony, M. capricolum subsp. capricolum, and bovine group 7 tested. Moreover, we found a nonspecific amplification product for M. putrefaciens that occurred with the oligonucleotide primers used for the "M. mycoides cluster" reaction. However, the restriction endonuclease digestion patterns observed with the restriction enzymes AluI, AseI, and SspI for M. putrefaciens were different than the digestion patterns obtained for the other "cluster" mycoplasmas. This report confirms the usefulness of polymerase chain reaction DNA amplification allied with restriction enzyme digestion profile analysis for the rapid and specific identification of mycoplasmas belonging to the "M. mycoides cluster" and M. putrefaciens. PMID- 9211240 TI - A monoclonal-antibody-based immunohistochemical method for the detection of swine influenza virus in formalin-fixed, paraffin-embedded tissues. PMID- 9211241 TI - Calicivirus outbreak with high mortality in a Missouri feline colony. PMID- 9211242 TI - Renal lesions associated with experimental porcine reproductive and respiratory syndrome virus (PRRSV) infection. PMID- 9211243 TI - Hepatitis and increased copper levels in a dalmatian. PMID- 9211244 TI - Alaria arisaemoides in a black Labrador retriever pup. PMID- 9211245 TI - A winter outbreak of anaplasmosis in a nonendemic area of Oklahoma: a possible role for Dermacentor albipictus. PMID- 9211246 TI - Acute clostridial hepatitis in an ostrich. PMID- 9211247 TI - Intraosseous hemangiosarcoma with metastasis in a three-month-old llama. PMID- 9211248 TI - Mineralization of the bulbus arteriosus in adult rainbow trout (Oncorhynchus mykiss). PMID- 9211249 TI - Chronic consumption of fumonisins derived from Fusarium moniliforme culture material: clinical and pathologic effects in swine. PMID- 9211250 TI - Effect of feeding deoxynivalenol (vomitoxin)-contaminated barley to horses. PMID- 9211251 TI - Analysis of ascorbic acid, dehydroascorbic acid, and transformation products by ion-pairing high-performance liquid chromatography with multiwavelength ultraviolet and electrochemical detection. PMID- 9211252 TI - Gas chromatographic/mass spectrometric measurement of ascorbic acid and analysis of ascorbic acid degradation in solution. AB - L-Ascorbic acid, DHA, and the oxidized products derived from AA can be accurately measured using GC/MS. Owing to the complex nature of the reactions through which AA proceeds, we believe that GC/MS is currently the procedure of choice in making AA-related measurements. The methods described are useful in defining reactions involving AA. The methods may indicate in vivo oxidative injury and may allow the use of AA-derived products to determine if antioxidant modulations are effective. PMID- 9211253 TI - Expression of recombinant L-gulono-gamma-lactone oxidase. PMID- 9211254 TI - Purification and characterization of glutathione-dependent dehydroascorbate reductase from rat liver. PMID- 9211255 TI - Peptidylglycine alpha-amidating monooxygenase: an ascorbate-requiring enzyme. PMID- 9211256 TI - Principles involved in formulating recommendations for vitamin C intake: a paradigm for water-soluble vitamins. PMID- 9211257 TI - High-performance liquid chromatography determination of total thiamin in biological and food products. PMID- 9211258 TI - Determination of thiamin and its phosphate esters in human blood, plasma, and urine. PMID- 9211259 TI - Determination of thiamin and thiamin phosphates in whole blood by reversed-phase liquid chromatography with precolumn derivatization. PMID- 9211260 TI - High-performance liquid chromatography determination of total thiamin in human plasma. PMID- 9211261 TI - Cyanogen bromide-based assay of thiamin. PMID- 9211262 TI - Isotopically labeled precursors and mass spectrometry in elucidating biosynthesis of pyrimidine moiety of thiamin in Saccharomyces cerevisiae. PMID- 9211263 TI - Thiamin transporters in yeast. PMID- 9211264 TI - In vitro systems for studying thiamin transport in mammals. PMID- 9211265 TI - Cofactor designing in functional analysis of thiamin diphosphate enzymes. PMID- 9211266 TI - Enzymatic preparation of derivatives of thiamin: O-beta-galactosylthiamin and O alpha-glucosylthiamin. PMID- 9211267 TI - High-performance liquid chromatography methods for determination of lipoic and dihydrolipoic acid in human plasma. AB - This assay method was applied to determine plasma levels of lipoic acid in humans. The method consists of enzymatic hydrolysis to release the protein-bound lipoic acid, solid-phase extraction, and electrochemical detection at a potential of +1.1 V. Previous methods did not provide adequate sensitivity for these studies or required procedural modifications for detection of low levels of plasma lipoic acid. The chromatographic system is capable of separating lipoic acid from dihydrolipoic acid. Both reduced and oxidized lipoic acid can be detected. Therefore, oxidation of dihydrolipoic acid must be prevented. In the described procedure, we do not prevent oxidation and the whole content is measured as lipoic acid. The method does not detect lipoic acid covalently bound to lysine. The detection limit for this method is 1 ng of lipoic acid per milliliter of plasma. PMID- 9211268 TI - Analysis of lipoic acid by gas chromatography with flame photometric detection. PMID- 9211269 TI - Biosynthesis of lipoic acid and posttranslational modification with lipoic acid in Escherichia coli. PMID- 9211270 TI - Lipoate addition to acyltransferases of alpha-keto acid dehydrogenase complexes and H-protein of glycine cleavage system. PMID- 9211271 TI - Lipoylation of acyltransferase components of 2-oxo acid dehydrogenase complexes. PMID- 9211272 TI - Purification and properties of brain lipoamidase. PMID- 9211273 TI - Measurement of pantothenic acid and hopantenic acid by gas chromatography-mass spectroscopy. PMID- 9211274 TI - Large-scale synthesis of coenzyme A esters. PMID- 9211275 TI - Synthesis of nonhydrolyzable acyl-coenzyme A analogs. PMID- 9211276 TI - Synthesis, purification, and characterization of dicarboxylylmono-coenzyme A esters. PMID- 9211277 TI - Holo-[acyl-carrier-protein] synthase of Escherichia coli. PMID- 9211278 TI - Determinations of biotin in biological fluids. PMID- 9211279 TI - Fluorophore-linked assays for high-performance liquid chromatography postcolumn reaction detection of biotin and biocytin. PMID- 9211280 TI - High-performance liquid chromatographic determination of biotin in biological materials after crown ether-catalyzed fluorescence derivatization with panacyl bromide. PMID- 9211281 TI - Bioluminescence competitive binding assays for biotin based on photoprotein aequorin. PMID- 9211282 TI - Competitive enzymatic assay of biotin. PMID- 9211283 TI - Competitive agglutination assay of biotin. PMID- 9211284 TI - Competitive enzyme-linked immunosorbent assay for biotin. PMID- 9211285 TI - Biotin synthesis in higher plants. PMID- 9211286 TI - Analysis of biotin biosynthesis pathway in coryneform bacteria: Brevibacterium flavum. PMID- 9211287 TI - Purification and characterization of biotin synthases. PMID- 9211288 TI - Biotin synthase of Bacillus sphaericus. PMID- 9211289 TI - Escherichia coli repressor of biotin biosynthesis. PMID- 9211290 TI - Structure of ATP-dependent carboxylase, dethiobiotin synthase. PMID- 9211291 TI - Purification and properties of bovine and human holocarboxylase synthetases. PMID- 9211292 TI - Biotin uptake in cultured cell lines. PMID- 9211294 TI - Biotinidase in serum and tissues. PMID- 9211293 TI - Biotinyl-5'-adenylate synthesis catalyzed by Escherichia coli repressor of biotin biosynthesis. PMID- 9211295 TI - Determination of biotinidase activity with biotinyl-6-aminoquinoline as substrate. PMID- 9211296 TI - Determination of serum biotinidase activity with radioiodinated biotinylamide analogs. PMID- 9211298 TI - Competitive binding assays for biotin-binding proteins. PMID- 9211297 TI - Preparation and properties of anti-biotin antibodies. PMID- 9211299 TI - Determination of vitamin B6 vitamers and metabolites in a biological sample. PMID- 9211300 TI - High-performance liquid chromatography determination of total pyridoxal in human plasma. PMID- 9211301 TI - Determination of 5-pyridoxic acid, 5-pyridoxic acid lactone, and other vitamin B6 compounds by cation-exchange high-performance liquid chromatography. PMID- 9211302 TI - Nuclear magnetic resonance in study of active sites of pyridoxal-dependent enzymes. PMID- 9211303 TI - Synthesis and application of pyridoxal polyphosphoryl derivatives as active-site probes for nucleotide-binding enzymes. PMID- 9211304 TI - Preparation of vitamin B6-peptide and vitamin B6-peptide-oligonucleotide conjugates. PMID- 9211305 TI - Preparation of nonlabeled, tritiated, and deuterated pyridoxine 5'-beta-D glucoside and assay of pyridoxine-5'-beta-D-glucoside hydrolase. PMID- 9211306 TI - Enzymatic preparation of pyridoxine 4'- and 5-alpha-D-glucosides. PMID- 9211307 TI - Formation of beta-galactosides of pyridoxine using Sporobolomyces singularis. PMID- 9211308 TI - Phosphatidylserine decarboxylases: pyruvoyl-dependent enzymes from bacteria to mammals. PMID- 9211309 TI - Production, assay, and occurrence of pyrroloquinoline quinone. PMID- 9211310 TI - Amine-oxidizing quinoproteins. PMID- 9211311 TI - Gas chromatographic-mass spectrometric analysis of pyrroloquinoline quinone. PMID- 9211312 TI - Monoclonal antibodies specific to pyrroloquinoline quinone. PMID- 9211313 TI - Determination, purification, and characterization of alpha-NADH and alpha-NADPH. PMID- 9211314 TI - Affinity labels for NAD(P)-specific sites. PMID- 9211315 TI - Photoaffinity labeling of NAD(+)-linked enzymes. PMID- 9211317 TI - Tissue NAD as a biochemical measure of niacin status in humans. PMID- 9211316 TI - Determining NAD synthesis in erythrocytes. PMID- 9211318 TI - Radioimmunoassay for measuring endogenous levels of cyclic ADP-ribose in tissues. PMID- 9211319 TI - Purification of human nicotinamide-mononucleotide adenylyltransferase. PMID- 9211320 TI - Nicotinamide-mononucleotide adenylyltransferases from yeast and other microorganisms. PMID- 9211321 TI - Use of biotinylated NAD to label and purify ADP-ribosylated proteins. AB - Biotin- or digoxigenin-conjugated NAD has been used successfully to label EF-2 by diphtheria toxin, an alpha subunit of G protein by pertussis toxin, and poly(ADP ribose) synthase through auto-poly(ADP-ribosyl)ation (J. Zhang, unpublished result, 1996). It is likely that many other ADP-ribosyl-transferases are capable of using modified NAD as substrates. Compared to radioactive labeling, biotinylation has several advantages. Commercially available precursors make synthesis of biotinylated NAD simple and economic. No extensive purification of the product is required. Because biotinylated NAD can be separated from NAD readily, there is no dilution, in contrast to [32P]NAD, in which only a small proportion of the NAD molecules are radioactive. Once purified, biotinylated NAD can be stored for a long time without decay (unlike radioactive NAD, which does decay). Most importantly, the system described here may afford an efficient means for purifying and identifying ADP-ribosylated proteins. Biotinylated NAD can be used for in situ labeling to study the cellular localization and tissue distribution of the ADP-ribosylated proteins. PMID- 9211322 TI - Preparation of cyclic ADP-ribose, 2'-phospho-cyclic ADP-ribose, and nicotinate adenine dinucleotide phosphate: possible second messengers of calcium signaling. PMID- 9211323 TI - Preparation of low molecular weight model conjugates for ADP-ribose linkages to protein. PMID- 9211324 TI - Bioassay for determining endogenous levels of cyclic ADP-ribose. PMID- 9211325 TI - Preparation of cyclic ADP-ribose antagonists and caged cyclic ADP-ribose. PMID- 9211326 TI - Synthesis and hydrolysis of cyclic ADP-ribose by human leukocyte antigen CD38: inhibition of hydrolysis by ATP and physiological significance. PMID- 9211328 TI - Large-scale purification of Aplysia ADP-ribosylcyclase and measurement of its activity by fluorimetric assay. PMID- 9211327 TI - Large-scale production of human CD38 in yeast by fermentation. PMID- 9211330 TI - Chemical synthesis and properties of 7 alpha-hydroxyriboflavin. PMID- 9211331 TI - Synthesis of N6-(2-aminoethyl)-FAD, N6-(6-carboxyhexyl)-FAD, and related compounds. PMID- 9211329 TI - Urinary riboflavin determination by C18 reversed-phase open-column chromatography. PMID- 9211332 TI - Biosynthesis of riboflavin: 3,4-dihydroxy-2-butanone-4-phosphate synthase. PMID- 9211333 TI - Biosynthesis of riboflavin: GTP cyclohydrolase II, deaminase, and reductase. PMID- 9211334 TI - Biosynthesis of riboflavin: lumazine synthase and riboflavin synthase. PMID- 9211335 TI - Measurements and characteristics of intestinal riboflavin transport. PMID- 9211336 TI - Purification and properties of FAD synthetase from liver. PMID- 9211338 TI - Purification and characterization of 5'-nucleotidase/FAD pyrophosphatase from human placenta. PMID- 9211337 TI - Determining covalent flavinylation. PMID- 9211339 TI - Syntheses and applications of flavin analogs as active site probes for flavoproteins. PMID- 9211340 TI - The hyperparathyroidism of chronic renal failure: a disorder of growth. PMID- 9211341 TI - Hyperhomocysteinemia in end-stage renal disease: prevalence, etiology, and potential relationship to arteriosclerotic outcomes. PMID- 9211343 TI - Autosomal dominant polycystic kidney disease with anticipation and Caroli's disease associated with a PKD1 mutation. Rapid communication. AB - Autosomal dominant polycystic kidney disease (ADPKD) is the most common renal hereditary disorder. Clinical expression of ADPKD shows interfamilial and intrafamilial variability. We screened for mutations the 3' region of the PKD1 gene, from exon 43 to exon 46, in a family showing anticipation and Caroli's disease and have found a 28 base pairs deletion in exon 46 (12801del28) and a new DNA variant in exon 43 (12184 C to G conserving Ala 3991) segregating with the disease. The mutation should result in a protein 44 amino acids longer then the wild-type PKD1. This PKD1 mutation manifests as typical adult-onset disease in the father, but in the proband, a 26-year-old man, ADPKD was diagnosed as a newborn and was associated with Caroli's disease at the age of 18 years. A renal biopsy performed in childhood disclosed a predominance of glomerular cysts. Mutation 12801del28 is the first molecular defect associated with Caroli's disease and the PKD1 phenotype. The finding of the same mutation in two different members of the same family with different expression of the disease indicates that the phenotypic variation in ADPKD must be due to modifying factors that may radically affect the course of the disease. PMID- 9211342 TI - Development of human fetal kidney in obstructive uropathy: correlations with ultrasonography and urine biochemistry. AB - In utero urethral obstruction results in bilateral hydronephrosis and severe fetal and post-natal morbidity and mortality. Obstetrical management depends on the indirect evaluation of fetal renal function by ultrasonography and biochemical analysis. No direct evaluation of the severity and possible reversibility of renal lesions is available. In this paper we analyzed kidneys from 34 fetuses (14 to 37 gestational weeks) in which (1) isolated bilateral urinary tract obstruction had been detected in utero by sonography, and (2) the severity of sonographic and biochemical prognostic indicators led to the indication of termination of pregnancy or to perinatal death. Pure hydronephrosis was observed in two young fetuses [14 and 20 gestational weeks (GW)] and was associated with regressive changes in two others. In contrast, a wide spectrum of dysplastic renal lesions was present in 30 fetuses and was classified into four subgroups according to the association of dysplasia, hypoplasia and cysts. They had the following characteristics in common: (1) premature cessation of nephrogenesis assessed by the medullary ray counting method; (2) early disappearance or myofibroblastic differentiation of metanephric blastema; (3) early increase in interstitial mesenchyme with widespread expression of alpha smooth muscle actin by mesenchymal cells; (4) frequent absence of classical criteria of dysplasia (nests of cartilage were observed in only 5 fetuses); (5) an identification, based upon the detection of alpha-smooth muscle actin expression, of the muscular phenotype of mesenchymal cells encircling primitive ducts. In conclusion, (1) the value of prognostic markers in fetuses less than 20 GW should be reconsidered; (2) after 20 GW there is a good correlation between markers predicting poor prognosis and the severity of renal lesions; (3) hypoplasia with disappearance of blastema cells, dysplasia and early interstitial fibrosis are evidence of the irreversibility of renal lesions and preclude any possibility of new nephron formation; (4) these findings suggest that most surgical in utero procedures are performed when irreversible renal lesions have developed. PMID- 9211344 TI - Effect of glyoxylate on the function of the calcitriol receptor and vitamin D metabolism. AB - The biological action of calcitriol is mostly mediated through the interaction of the calcitriol receptor (VDR) with vitamin D response elements (VDREs) of target genes. These interactions produce special proteins that carry out the biological activities of calcitriol. Recently, we showed that the interaction of VDRs with VDREs is inhibited by uremic toxins. We hypothesize that uremic toxins that contain aldehyde or ketone groups potentially could form Schiff bases with lysine residues of the VDR DNA binding domain and inhibit VDR interaction with VDREs. We therefore chose glyoxylate, a compound which has an aldehyde group, to test this hypothesis. In vitro glyoxylate inhibited VDR binding to the osteocalcin and osteopontin VDREs as assessed by electrophoretic mobility shift assay and the inhibition was reversed when glyoxylate was preincubated with lysine. Further, this chemical compound also blocked the induction of chloramphenicol acetyltransferase (CAT) enzyme induced by calcitriol in cells transfected with a calcitriol responsive CAT reporter gene. Since induction of 24-hydroxylase synthesis is a VDR regulated process, we also studied the effect of glyoxylate on the activity of intestinal 24-hydroxylase in rats. This enzyme activity was suppressed in rats infused with glyoxylate. Taken together, our study suggests that glyoxylate could inhibit the interaction of VDR with VDREs and alter the biological action of calcitriol. PMID- 9211345 TI - Parathyroid hormone prevents 1,25 (OH)2D3 induced down-regulation of the vitamin D receptor in growth plate chondrocytes in vitro. AB - 1,25(OH)2D3 has an antiproliferative effect on growth plate chondrocytes when given in high doses, whereas low doses stimulate chondrocyte proliferation. In the present in vitro study we investigated the effects of parathyroid hormone (PTH) when given concomitantly with 1,25(OH)2D3 on cell proliferation and vitamin D receptor (VDR) regulation. Freshly isolated rat tibial chondrocytes were grown in monolayer cultures or in agarose stabilized suspension cultures (10% charcoal treated FCS). VDR expression was determined by RT-PCR generating a 297 bp fragment and by binding assays (Scatchard analysis) with [3H]-1,25(OH)2D3. Cell proliferation was measured by [3H]-thymidine incorporation, growth curves in monolayer cultures and by colony formation in agarose-stabilized suspension cultures. Optimal concentration of 1,25(OH)2D3 (10(-12) M) and of PTH fragments [bPTH(1-34) or hPTH(28-48), 10(-10)M] showed additive effects on DNA synthesis of and colony formation by growth plate chondrocytes. This may be explained in part by an up-regulation of VDR by PTH: PTH increased both mRNA expression of VDR and binding capacity. 1,25(OH)2D3 (10(-12) M) induced an up-regulation of the VDR within 24 hours followed by a down-regulation after incubation for more than 24 hours. PTH fragments added concomitantly prevented the down-regulation seen with 1,25(OH)2D3. These findings provide evidence that PTH is a growth promoting hormone that also modulates the effects of 1,25(OH)2D3 by regulating the VDR status of 1,25(OH)2D3 target cells. PMID- 9211346 TI - Interleukin-4 and interleukin-10 attenuate established crescentic glomerulonephritis in mice. AB - Crescentic glomerulonephritis (GN) has immunopathological features of delayed type hypersensitivity (DTH) and results from a T helper cell 1 (Th1) dependent immune response. The current study examined the capacity of Th2 cytokines, interleukin (IL)-4 and IL-10, to alter the outcome of crescentic GN, after injury is established. Sensitized, control treated mice developed crescentic GN with functional renal injury (117 +/- 20 microliters/min, normal mouse 182 +/- 8 microliters/min, P < 0.05) 10 days after an i.v. dose of sheep anti-mouse glomerular basement membrane globulin. Combined treatment with IL-4 and IL-10 starting three days after initiation of disease significantly reduced glomerular crescent formation (5.3 +/- 3.2%, control treatment 23.3 +/- 6.4%, P < 0.02) and preserved renal function (165 +/- 15 microliters/min, P = 0.57 compared to normal mice). Treatment with IL-4 alone did not reduce crescent formation or protect renal function. Mice treated with IL-10 showed trends to decreased crescent formation and preservation of renal function. In all cytokine treated groups, the accumulation of effectors of glomerular injury (CD4+ positive T cells, macrophages and fibrin) was reduced, with the combination treatment having the greatest effect. Administration of Th2 cytokines, IL-4 and IL-10 to mice with established GN attenuates the development of glomerular crescent formation and protects renal function. PMID- 9211347 TI - Altered ceramide and sphingosine expression during the induction phase of ischemic acute renal failure. AB - Recent evidence indicates that a "sphingomyelin signaling pathway" exists: in response to heterogeneous influences, sphingomyelin is hydrolyzed, liberating ceramide, and subsequently its sphingoid base, sphingosine. Ceramide and sphingosine can influence diverse cellular processes, including cell differentiation, proliferation, protein trafficking, and apoptosis. Each of these processes have important implications for post-ischemic acute renal failure (ARF). However, sphingosine and ceramide expression during the induction of ischemic/reperfusion injury have not been previously assessed. To this end, CD-1 mice were subjected to 45 minutes of unilateral renal ischemia +/- reperfusion, followed by cortical sphingosine, ceramide, and sphingomyelin assessments. Contralateral kidneys served as controls. Ischemia caused approximately 50% sphingosine and ceramide decrements. During reperfusion, sphingosine rebounded to normal values. Conversely, ceramide rose to, and was maintained at, supranormal levels (approximately 175% of controls). Subsequent studies performed with hypoxic or oxygenated isolated proximal tubules suggested that these changes: (1) had a multifactorial basis; (2) were partially simulated by enhanced PLA2 activity; (3) and were dissociated from alterations in net sphingomyelin content. To assess the potential pathogenic relevance of the documented ceramide increments, cultured human proximal tubule (HK-2) cells were subjected to ATP depletion/Ca2+ ionophore- or PLA2-induced attack with or without exogenous C2 ceramide loading. Ceramide worsened both forms of injury without exerting an independent lethal effect. Conversely, ceramide markedly attenuated arachidonic acid cytotoxicity. This occurred without any decrease in arachidonate uptake, suggesting a direct cytoprotective effect. IN CONCLUSION: (1) sphingosine and ceramide fluxes are hallmarks of early ischemic/reperfusion injury; (2) these changes occur via divergent metabolic pathways; and (3) that ceramide increments can affect divergent injury pathways, and that sphingosine and ceramide have potent cell signaling effects, suggest that the currently documented sphingosine/ ceramide fluxes could have important implications for the induction phase and evolution of post-ischemic ARF. PMID- 9211348 TI - Maturational changes in rabbit renal cortical phospholipase A2 activity. AB - Several studies have demonstrated that the neonatal kidney has a markedly attenuated response to parathyroid hormone (PTH); however, the cause for this blunted response is unknown. PTH stimulated cAMP production by 215 +/- 18% in neonatal proximal tubule suspensions compared to a 35 +/- 7% increase in adult proximal tubules. Thus, neonatal proximal tubules have functioning PTH receptors and a greater adenylate cyclase response than the adult segment. In adult proximal tubules, PTH stimulates phospholipase A2 (PLA2) activity and the inhibition of Na,K-ATPase activity by PTH is blocked by inhibitors of PLA2. We examined whether maturational changes in renal cortical activity could play a role in the attenuated response to PTH in the neonatal proximal tubule. Compared to adults, neonates had a lower renal cortical cytosolic PLA2 (cPLA2) activity, assessed as the release of 14C-arachidonic acid (AA) from labeled phosphatidyl choline (0.44 +/- 0.10 vs. 0.74 +/- 0.06% 14C-AA released/min/mg protein, P < 0.05) and microsomal PLA2 activity (0.32 +/- 0.03 vs. 1.20 +/- 0.13% 14C-AA released/min/mg protein, P < 0.001). The protein abundance of cPLA2 was not different between the neonatal and adult renal cortex as assessed by immunoblot assay. Thus, the difference in activities must be due to a difference in regulation of cPLA2. Annexin 1 (lipocortin 1) has been shown to inhibit PLA2 activity by binding to phospholipid substrate. Annexin 1 protein abundance was higher in neonatal than in adult renal cortex (P < 0.001). Thus, the lower activity of PLA2 in the neonatal tubules may be due in part to higher expression of annexin 1. PLA2 activation by PTH, -8-bromo-cAMP and PMA was assessed as 3H-AA release from prelabeled suspensions of neonatal and adult proximal tubules. PTH (10(-7) M), 8-bromo-cAMP (10(-4) M) and PMA (5 x 10(-8) M) significantly increased 3H-AA release from adult tubules (P < 0.05) but had no effect on neonatal tubules (P = NS). Thus, PTH, 8-bromo-cAMP and PMA stimulated PLA2 in adult but not neonatal proximal tubules. In conclusion, the maturational changes in renal cortical PLA2 activity may be a factor in the blunted response of neonatal proximal tubules to PTH. PMID- 9211349 TI - Effect of glucose on the function of the calcitriol receptor and vitamin D metabolism. AB - The genomic action of calcitriol is mediated through the interaction of the calcitriol receptor (VDR) with vitamin D response elements (VDREs) of the target genes. It has been proposed that chemicals capable of Schiff base formation with the VDR potentially could alter the physiological function of VDR and calcitriol metabolism. Since glucose has been shown to form Schiff bases with proteins, we tested the hypothesis that glucose could influence the function of VDR and thereby alter calcitriol metabolism. Glucose 6-phosphate inhibited VDR binding to the osteocalcin VDRE and chemically modified the DNA binding domain or the dimerization domain of the VDR in vitro. Further, glucose also blocked the production of chloramphenicol acetyltransferase (CAT) enzyme induced by calcitriol in cells transfected with a constructed VDRE attached to a CAT reporter gene. Hyperglycemia induced by glucose infusion or by streptozotocin in normal rats significantly reduced intestinal 1 alpha, 25-dihydroxyvitamin D-24 hydroxylase activity. Taken together, these findings are consistent with the hypothesis that glucose could interact with the VDR to impair its DNA binding and function within cells. PMID- 9211350 TI - Effect of glucose, pyruvate, and insulin on type 1 angiotensin II receptor expression in SV40-immortalized rabbit proximal tubule epithelial cells. AB - Ambient glucose concentrations alter type 1 angiotensin II receptor (AT1R) expression in renal tissues. The direction of change in AT1R density may depend on the specific cell type and the capacity for that cell type to use glucose as an energy substrate. Given the effects of angiotensin II (Ang II) in proximal tubule epithelia, glucose-mediated fluctuations in AT1R expression could significantly alter tubular Na(+)-H+ exchange and volume reabsorption. To determine if glucose influenced AT1R expression in cultured proximal tubule epithelial cells, SV40-immortalized rabbit proximal tubule epithelial cells (RPTEC) were exposed to 25 mmol (hi-glc) or 5 mmol glucose-containing serum-free medium (lo-glc) for seven to nine days, with or without an alternative energy substrate, pyruvate. AT1R expression, assessed by quantitative reverse transcription polymerase chain reaction and specific 125I-Ang II binding, decreased in lo-glc medium (% reduction AT1R mRNA expression: 52 +/- 8%; N = 6; P < 0.005 vs. hi-glc; % reduction specific 125I-Ang II binding: 48 +/- 12%; N = 12; P < 0.03 vs. hi-glc). AT1R mRNA expression and specific 125I-Ang II binding recovered to hi-glc levels following the addition of pyruvate [60 mmol] to lo-glc cells. To ascertain if a growth factor that increases glucose uptake in vivo also altered AT1R expression, RPTEC were cultured in hi-glc medium with or without exogenous insulin [100 nM]. Insulin addition increased AT1R mRNA expression and specific 125I-Ang II binding in a concentration-dependent manner. However, insulin (100 nM) addition to lo-glc cells did not significantly increase specific 125I-Ang II binding. These results suggest that AT1R expression in SV40 immortalized rabbit proximal tubule cells is significantly affected by the availability of energy substrate. Ultimately, changes in proximal tubule AT1R expression, mediated by elevated glucose concentrations and insulin, could contribute to sodium-dependent hypertension in the setting of hyperinsulinemia and hyperglycemia. PMID- 9211351 TI - Two novel probes reveal tubular and vascular Arg-Gly-Asp (RGD) binding sites in the ischemic rat kidney. AB - We have previously demonstrated that RGD peptides prevent tubular obstruction in ischemic acute renal failure (ARF) suggested that exposed unoccupied integrin receptors represent the target for such therapy. The present study investigated the topography of RGD binding sites and integrin receptors in ischemic rat kidneys. Two RGD peptides were synthesized: a cyclic biotinylated (Bt) RGD peptide and a linear RGD peptide (GRGDSP) labeled with rhodamine green (RhoG). Rats were subjected to 45 minutes of renal artery occlusion kidneys were harvested at different times post-ischemia, and stained with RGD peptides and a panel of antibodies to integrins. In control, Bt-RGD staining was undetectable in alkaline phosphatase histochemistry, whereas immunofluorescence detection with Rho-streptavidin conjugate as well as RhoG-GRGDSP staining faintly decorated the basolateral aspect of the proximal tubular cells in a punctate fashion. In contrast, ischemic kidneys showed binding to the basolateral and apical aspects of proximal tubules, peritubular capillaries, and desquamated cells within tubular lumen. The most conspicuous staining of ischemic kidneys was obtained with antibodies to the beta 1 (labeling of the apical aspect of proximal and distal tubules, as well as desquamated cells obstructing tubular lumen) and the alpha V (glomeruli, tubular epithelia, intima of blood vessels stained faintly, while the obstructing cellular conglomerates showed intense staining) subunits. Double staining with Bt-RGD and antibodies against the beta 1 and alpha V beta 3 integrins showed co-localization of staining within the tubules and vasculature, respectively. In vitro attachment of HL-60 leukocytes to the endothelial cells was inhibited by the cyclic RGD peptide. In conclusion, expression of RGD binding sites and beta 1 integrin subunits along the apical aspect of tubular epithelia and on the surface of desquamated cells is in concert with the hypothesis on the pathogenetic role of RGD-recognizing integrins in tubular obstruction. The expression of RGD binding sites along the intimal surface of blood vessels in ischemic kidneys suggests an additional target for RGD peptides in vascular endothelial cells. PMID- 9211352 TI - Importance of the tubulointerstitium in human glomerulonephritis. II. Distribution of integrin chains beta 1, alpha 1 to 6 and alpha V. AB - Accumulation of extracellular matrix is important in the progression of glomerulonephritis. Since adherent cell types utilize integrins to bind and organize extracellular matrix proteins, we have assessed expression of the beta 1 integrins in sequential sections from 85 human renal biopsies and 4 normal kidneys by immunohistochemical staining. Our results demonstrate strong correlations between expression of the alpha 5 chain within the interstitium, the alpha V chain on proximal and distal tubular epithelium and the presence of chronic histological damage. Moreover, staining for interstitial alpha 5 and proximal and distal tubular alpha V were also strongly associated with expression of certain adhesion molecules (ICAM-1, VCAM-1, E-selectin and L-selectin) and the presence of macrophages within the interstitium, which have been linked, in an earlier study, with the degree of chronic histological damage and disease progression. However, in contrast to our earlier study of adhesion molecules, there were also associations between expression of integrin chains within the glomerulus and tubulointerstitium. For example, there were strong positive associations between staining for alpha 5 on glomerular endothelium and its expression on extraglomerular vascular endothelium and between both mesangial alpha 1 and podocyte alpha 3 and tubular staining for the common beta 1 subunit. While the functional significance of these associations is obscure, they suggest some kind of communication between cells in different sites in the kidney. There were also positive associations between staining for different integrins within the glomerulus, notably mesangial cell staining for alpha 2, glomerular endothelial cells staining for alpha 5 and glomerular epithelial cell alpha 3. These results suggest that there is a coordinated upregulation of integrin expression both within the tubulointerstitium and the glomerulus and that at least some of these integrins (interstitial alpha 5 and distal tubular alpha V) are associated with the expression of other adhesion molecules, macrophage infiltration and the presence of markers of disease progression (interstitial fibrosis and tubular atrophy). PMID- 9211353 TI - In situ hybridization studies of matrix metalloproteinase-3, tissue inhibitor of metalloproteinase-1 and type IV collagen in diabetic nephropathy. AB - Progressive expansion of the mesangial matrix is one of the most characteristic histological features of diabetic nephropathy (DN). To determine the balance between the turnover and degradation of extracellular matrix (ECM) in renal tissue of patients with DN, we examined the expression of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1) and type IV collagen (IV-C) mRNAs using a high-resolution in situ hybridization. Patients were divided into three grades: mild (grade I), moderate (grade II) and severe (grade III) mesangial expansion and tubulointerstitial injury. The relationship between the expression of these mRNAs and degree of glomerular mesangial expansion and interstitial injury was also examined. Cells positive for each mRNA were observed in glomerular resident cells, including glomerular mesangial, epithelial and endothelial cells and cells of Bowman's capsule. A number of tubular epithelial cells and some infiltrating cells in the interstitium also expressed these mRNAs. The expression of MMP-3 mRNA and TIMP-1 mRNA was strongest in glomeruli of grade I and inversely correlated with mesangial expansion. In contrast, the expression of all three types of mRNA was correlated with the degree of interstitial injury. Our results indicate that IV C, MMP-3 and TIMP-1 mRNAs are expressed in glomerular resident cells, tubular epithelial cells and infiltrating cells in renal tissue of DN, and suggest that their expression changes with the degree of mesangial expansion and interstitial injury. Altered expression of MMP-3 and TIMP-1 may be associated with the progression of DN. PMID- 9211354 TI - Identification of promoter activity and differential expression of transcripts encoding the murine stromelysin-1 gene in renal cells. AB - Stromelysin-1, matrix metalloproteinase-3 (MMP-3), is an important endopeptidase selectively expressed by somatic cells in organ tissues. The renal tubulointerstitium, for example, comprises tubular epithelium and interstitial fibroblasts forming the principal mass of the kidney. We observed that mRNA encoding stromelysin-1 is detectable in murine renal fibroblasts, but not in proximal tubular epithelium. Transcripts measured by RNase protection assay in renal fibroblasts increase following exposure to phorbol ester, and thereafter, activated stromelysin-1 protein can be detected in culture media by Western blotting. A 6.4 Kb genomic clone containing the putative stromelysin-1 promoter was isolated and a relevant 2.1 Kb PstI restriction fragment including 2.1 Kb of the immediate 5'-flanking region was sequenced on both strands. Two transcriptional start sites were identified by primer extension; the major start site corresponded to a previously established position in the rat promoter, and a second undescribed minor transcriptional start site was located 16 bp upstream of the primary site. A HiNF-A chromatin-activating element at -106 bp was found in the early promoter region of pR336 and an active AP-1 site at -72 bp with an Ets/PEA-3 motif at -203 bp was suggested by transient transfection of luciferase minigenes into renal fibroblasts responsive to phorbol ester. This Ets element was identical to a site in the early promoter of the fibroblast-specific gene FSP1. A baseline enhancement in activity of pR336 in fibroblasts was further observed with the addition of 5' flanking sequence out to -1980 bp. This additional region of flanking sequence contains two modular regions: one of multiple PEA-3 elements between -684 bp and -1955 bp and a second region between 1929 bp and -1980 bps containing a second AP-1 site at -1929 bp, a MBF-1/ MEP-1 metal binding site, and a PPAR peroxisome proliferator element at -1950 bp. Our findings implicate a gene structure with expected activity in a mesenchymal phenotype. The PKC-dependent regulation of the stromelysin-1 gene supports the notion that it may be modulated during inflammation or tissue remodeling. PMID- 9211355 TI - Modulation of c-fos and egr-1 expression in the isolated perfused kidney by agents that alter tubular work. AB - The isolated perfused rat kidney provides a model of selective hypoxia to the medullary thick ascending limb. To investigate the relationship between immediate early gene expression and the extent of hypoxic damage, we determined expression of the immediate early genes (IEG) c-fos and egr-1 in isolated perfused kidneys during standard perfusion and after various measures shown previously to be protective. mRNA levels of c-fos and egr-1 were markedly increased in kidneys after 90 minutes of standard perfusion with Krebs-Henseleit buffer containing albumin. Gene expression was most prominent in the outer medulla followed by papilla and cortex, a pattern reflected by the immunohistochemical demonstration of a prominent accumulation of both egr-1 and c-fos polypetides mainly in the medullary thick ascending limb (mTAL). Protective measures known to minimize morphological damage to the mTAL, including hyperoncotic perfusion, perfusion with glycine, or perfusion with a mixture of amino acids, decreased mRNA levels of c-fos and egr-1 in the outer medulla (by 50% and 35%, respectively) and the papilla (by 60 and 30%, respectively). Renal cortex showed only minor changes. In contrast, prevention of tubular transport by perfusion with 1 mM ouabain increased mRNA levels of c-fos and egr-1 in the outer medulla by 100% and 60%, respectively. Ouabain also dramatically increased mRNA levels of both IEGs in two lines of cultured renal epithelial cells. Changes in the level and distribution of the protein products of these IEGs were not detectable in perfused kidneys by immunohistochemistry. Hypoxic injury of the kidney stimulates IEG expression even in the absence of reperfusion. Protection against hypoxic injury in the mTAL correlates with suppression of IEG mRNA levels when protection is provided by amino acids or hyperoncotic perfusion, but not when provided by inhibition of Na,K-ATPase, which stimulates IEG expression. We conclude that diminished IEG expression is not a necessary concomitant of protection against hypoxic injury. PMID- 9211356 TI - Glycine-protected, hypoxic, proximal tubules develop severely compromised energetic function. AB - Glycine-treated, hypoxic, proximal tubules developed a progressive energetic defect that resulted in failure to restore ATP levels to greater than 10 to 20% of control values during reoxygenation after 60 minutes of hypoxia despite continued cytoprotection by glycine. The defect was not corrected by supplementation with exogenous purines and was not modified by lowering the pH during hypoxia or reoxygenation. In the continued presence of glycine, the failure to restore ATP was associated with impaired recovery of structural changes that developed during hypoxia and, if glycine was withdrawn, lethal membrane damage occurred. The lesion was significantly ameliorated by the presence during hypoxia of two agents known to suppress development of the mitochondrial permeability transition, cyclosporine A and butacaine, which were most effective when used in combination. The data suggest that development of the mitochondrial permeability transition in glycine-protected tubules during hypoxia contributes to continued metabolic and structural impairment and cell death that occur despite glycine replete conditions such as exist frequently during in vivo insults and may be a target for therapeutic maneuvers. PMID- 9211357 TI - Opposite paracrine effects of 5-HT and dopamine on Na(+)-Pi cotransport in opossum kidney cells. AB - Serotonin (5-HT) was recently reported to inhibit cAMP generation in oppossum (OK) cells. We thus investigated the effects of 5-HT upon the Na(+)-Pi cotransport in cultured OK cells and its interactions with dopamine. Incubation of OK cells with 1 nM-10 microM 5-HT resulted in dose-dependent stimulation of Na(+)-Pi contransport (ED50 approximately equal to 8 nM) and also counteracted inhibition of Na(+)-Pi cotransport elicited by dopamine. Pre-incubation with 5-HT decreased cAMP accumulation elicited by forskolin or dopamine and pre-treatment with pertussis toxin abolished both the inhibitory effect of 5-HT upon cAMP levels and stimulation of Na(+)-Pi cotransport. Incubation of OK cells with the 5 HT precursor 5-hydroxytryptophan resulted in time- and dose-dependent accumulation of 5-HT in the medium that also elicited an increase in Na(+)-Pi cotransport. Both the effects of 5-HT and dopamine on Na(+)-Pi cotransport were prevented by carbidopa. The stimulatory effect of 5-HT was specific for the Na(+) Pi cotransport system since no effects were observed on Na(+)-alanine cotransport. The results indicate that 5-HT stimulates Na(+)-Pi cotransport at least in part via inhibition of cAMP accumulation. We propose that 5-HT and dopamine have opposite actions as paracrine/autocrine regulators of Na(+)-Pi cotransport via opposite effects upon cAMP formation. PMID- 9211359 TI - Modulation of renal hemodynamics by IGF-1 is absent in spontaneously hypertensive rats. AB - We recently reported that attenuation of vasoactive agent-induced calcium signal and cell contraction of mesangial cell by insulin-like growth factor 1 (IGF-1), observed in normal mesangial cells, is totally abolished in spontaneously hypertensive rat (SHR) mesangial cells. This phenomenon might be related to the well-known aberrant regulation of SHR glomerular hemodynamics. Since it has been reported that in vivo IGF-1 infusion increases renal plasma flow (RPF) and glomerular filtration rate (GFR), we examined whether the modulation of renal function by IGF-1 is altered in SHR. We performed in vivo renal clearance studies using eight-week-old SHR and control Wistar Kyoto rats (WKY) before and after IGF 1 (5 micrograms/kg) infusion into the left renal artery for 20 minutes. Mean arterial pressure was not affected by IGF-1 in both WKY and SHR. In WKY, IGF-1 increased GFR and RPF, and decreased renal vascular resistance (RVR). However, GFR, RPF, and RVR were not altered by IGF-1 in SHR, while systemic infusion of angiotensin II antagonist, CV-11974, increased GFR and RPF. The present data show that the modulation of renal hemodynamics by IGF-1 is absent in SHR. This might be related the pathophysiology of the development of hypertension. PMID- 9211358 TI - Agonist-induced activation of a non-selective ion current in glomerular endothelial cells. AB - The control of intracellular calcium activity ([Ca2+]i) and membrane voltage (Vm) play an important role in regulating functions of glomerular endothelial cells (GEC). We investigated the effect of extracellular ATP on the intracellular [Ca2+]i, Vm and ion conductances in GEC. ATP (100 mumol/liter) induced a rapid increase of [Ca2+]i in GEC from 20 +/- 6 to 442 +/- 84 nmol/liter, which was followed by a sustained Ca2+ plateau of 112 +/- 29 nmol/liter. In a bath solution with a low extracellular Ca2+ concentration the ATP-induced [Ca2+]i peak was still present, but the [Ca2+]i plateau was completely prevented. In 186 experiments with the patch clamp technique the addition of ATP (1 to 100 mumol/liter) to GEC induced a transient small hyperpolarization, which was followed by a depolarization. During the ATP-induced depolarization an increase of the whole cell conductance was found. The Ca2+ ionophore A23187 (10 mumol/liter) mimicked the effect of ATP on Vm. Reduction of the extracellular Ca2+ to 1 mumol/liter itself depolarized GEC reversibly from -88 +/- 2 to -60 +/- 12 mV and increased the ATP-induced depolarization to -18 +/- 3 mV. In the absence of Na+ in the bathing solution (replacement by NMDG+) ATP induced only an attenuated depolarization and no inward current was activated. Flufenamate (100 mumol/liter), a blocker of non-selective ion channels inhibited the ATP-induced depolarization of Vm significantly by 58 +/- 13%, whereas nicardipine (10 mumol/liter) or amiloride (10 mumol/liter) had no effect. Our data indicate that the resting Vm of GEC cells is almost completely dominated by K+ conductances and that ATP activates a Ca2+ dependent non-selective ion conductance in GEC. PMID- 9211360 TI - Renal and systemic nitric oxide synthesis in rats with renal mass reduction. AB - In rats undergoing renal mass reduction (RMR) oral supplementation with the nitric oxide (NO) precursor L-arginine increases glomerular filtration rate and ameliorates signs of glomerular injury, suggesting that chronic renal failure in the rats is a condition of low NO formation in the kidney. On the contrary, data are available that in the systemic circulation of uremics, both rats and human beings, NO is formed in excessive amounts and may contribute to platelet dysfunction and bleeding tendency, well-known complications of uremia. The present study was designed to clarify the pathophysiology of renal and systemic NO synthesis in uremia. We showed that renal ex vivo NO generation, measured as the conversion of [3H] L-arginine to [3H] L-citrulline, was lower than normal in RMR rats, seven days after surgery, and progressively worsened with time in close correlation with signs of renal injury. Consistent with these results, urinary excretion of the stable NO metabolites, NO2-/NO3-, significantly decreased in rats with RMR. To go deeper into the cellular origin and biochemical nature of this abnormality we used two histochemical approaches that could locate either NO synthase (NOS) catalytic activity (NADPH-diaphorase) or NOS isoenzyme expression (immunoperoxidase). NADPH-diaphorase documented a progressive loss of renal NOS activity in RMR rats that co-localized with a strong progressive decrease of inducible NOS isoenzyme (iNOS) immunostaining. At variance with iNOS, endothelial cell NOS (ecNOS) staining was rather comparable in RMR and control kidneys. At variance to the kidney, in the systemic circulation of RMR rats the synthesis of NO increased as reflected by higher than normal plasma NO2-/NO3- concentrations. High systemic NO likely derives from vessels as documented by the increased NOS activity and higher expression of both iNOS and ecNOS in the aorta of RMR rats. Up-regulation of systemic NO synthesis might be an early defense mechanism against hypertension of uremia. On the other hand, more NO available to circulating cells may sustain the bleeding tendency, a well-known complication of uremia. PMID- 9211361 TI - A new model of renal microvascular endothelial injury. AB - Although the importance of injury with consequent activation of endothelium is well-recognized in diseases affecting the glomerular endothelial cell (GEN), research on GEN injury in vivo has been hampered by the lack of adequate animal models. Here we report the establishment and characterization of a new GEN injury model in rats. This model was induced by selective renal artery perfusion with anti-GEN IgG and resulted in the severe acute renal failure with marked platelet deposition and development of a thrombotic microangiopathy involving glomeruli. Peritubular capillary endothelial cells were also damaged that was associated with severe tubular necrosis. Although the glomerular changes were severe, half of the glomeruli recovered by day 10, while interstitial changes remained throughout our observation time course. Proliferation of GEN was observed during the recovery phase. An increased expression of endothelial nitric oxide synthase in GEN was also observed, and may be an adaptive mechanism to counteract the thrombosis and ischemia. This model should be useful to investigate the pathophysiology of renal microvascular diseases and the mechanisms of GEN injury, activation and recovery in vivo. PMID- 9211362 TI - Up-regulation of renal and vascular nitric oxide synthase in iron-deficiency anemia. AB - Anemia is frequently associated with increased cardiac output and reduced vascular resistance. The latter has been attributed to reduced inactivation of nitric oxide (NO) by hemoglobin. We hypothesized that in addition to reducing NO inactivation, anemia may up-regulate NO production. To test this hypothesis, male Sprague-Dawley rats with chronic iron-deficiency anemia (produced by multiple phlebotomies and an iron-free diet) were studied. The results were compared with those obtained in a group of normal control animals. The anemic group showed marked increases in urinary excretion, plasma concentration, and renal and aorta tissue contents of NO metabolites (total nitrates and nitrites, NOx). This was accompanied by a significant rise in urinary excretion of cGMP, the second messenger for NO. In addition, NO synthase (NOS) activity and endothelial constitutive (ecNOS) and inducible NOS (iNOS) proteins of the thoracic aorta were markedly increased in the anemic group. Likewise, renal tissue ecNOS and iNOS proteins were greatly increased in the anemic animals. NOS activity and protein values were inversely related to hematocrit and directly related to plasma, tissue and urinary NOx. The constellation of these findings points to an increased NOS expression and NO production as opposed to the mere reduction of NO inactivation in iron-deficiency anemia. Further studies are planned to determine the mechanism of NOS up-regulation in iron-deficiency anemia. PMID- 9211363 TI - Natriuretic peptide receptors mediate different responses in rat renal microvessels. AB - Atrial natriuretic peptide (ANP) has unique effects on the renal vasculature, in that it dilates preglomerular vessels and constricts efferent arterioles. In the present study we aimed to characterize the natriuretic peptide receptor (NPR) subtypes, which mediate the renovascular effects of ANP, using in vivo microscopy in the split hydronephrotic kidney model of rats. ANP (10(-9) and 3.10(-9)), which binds to NPR-A and NPR-C, dilated preglomerular vessels and constricted efferent arterioles similarly to that found in previous studies. C-type natriuretic peptide (10(-9) to 10(-7)), which binds to NPR-B and NPR-C, dilated pre- and postglomerular vessels and profoundly increased glomerular blood flow. A specific ligand of NPR-C, C-ANP (des-[Gln18,Ser19,Gly20,Leu21,Gly22]ANP 4-23-NH2, 10(-9) to 10(-7)) was devoid of vascular effects. The ANP antagonist A71915 (10( 9) to 10(-6)) induced moderate dilation in renal vessels possibly due to some agonistic activity on NPR-B, ANP-induced preglomerular vasodilation was attenuated by A71915 (10(-6)) to 36 +/- 6% of the initial response, whereas efferent vasoconstriction was completely abolished (-4 +/- 4% of initial response). Our results indicate that ANP dilates preglomerular vessels and constricts efferent arterioles through NPR-A, both responses being antagonized by A71915 with different potencies. Furthermore, our data show that in the rat renal microcirculation stimulation of NPR-B results in vasodilation only, whereas NPR-C does not mediate vascular responses. PMID- 9211364 TI - Recurrent corneal erosion associated with Alport's syndrome. Rapid communication. AB - Ocular defects associated with Alport syndrome (AS) include anterior lenticonus and retinal flecks. We report on recurrent corneal erosion (RCE) as another ocular manifestation of the disease. Three brothers with AS reported a history of spontaneous attacks of RCE (2 episodes over 1 to 3 years in 2 of them and about 60 episodes in one brother over the last 10 years) characterized by acute ocular pain, lacrimation and photophobia lasting two to five days. The absence of RCE in the two other non-affected brothers from the same kindred suggested an association between AS and RCE, and prompted us to assess its prevalence. Forty one patients with AS and renal failure and 67 control transplanted patients (with another original nephropathy) were evaluated. Seven AS patients had a history of RCE (first manifested between the ages of 12 and 21) versus only one control patient (P = 0.003). In conclusion, a history of RCE is found in about 20% of patients with AS and renal failure. RCE is likely to result from an inherent structural weakness of the corneal epithelial basement membrane (containing type IV collagen). A history of RCE should be sought when evaluating a patient for AS. Ophthalmologists should also be aware of this association, when confronted with a patient suffering from non-traumatic RCE. PMID- 9211365 TI - Low efficiency of active immunization against diphtheria in chronic hemodialysis patients. AB - Recent epidemiological studies indicate a low immunity to diphtheria in adults in industrialized countries. In the light of the epidemic increase of diphtheria in countries such as Russia and the Ukraine, systematic vaccination against this disease is recommended. We analyzed the immunity to diphtheria of 228 hemodialysis patients and the efficiency of single versus triple vaccination against diphtheria. Antibodies against diphtheria toxoid were determined by enzyme immunoassay in sera of 228 adult hemodialysis patients. Fifty-four patients were triple vaccinated against diphtheria and were followed for six months; 17 patients were single immunized and antitoxoid titers were determined 1 and 12 months later. The overall protection rate against diphtheria was 22% and equal in male and female patients. After triple immunization, only 35% of the patients developed protective antibody concentration (> 0.1 i.e./ml) six months after the third vaccination. A single vaccination caused protective titres twelve months later in 41% of the patients. There was no difference between responders and non-responders in the duration, intensity or modality of hemodialysis treatment or the response to previous vaccinations against hepatitis-B. We suggest to monitor antibodies against diphtheria toxoid in vaccinated hemodialysis patients at risk for diphtheria since protective titers are often not attained by the standard vaccination protocol. PMID- 9211366 TI - Reticulocyte hemoglobin content in the evaluation of iron status of hemodialysis patients. AB - The assessment of iron status for hemodialysis patients has been hindered by the inaccuracy of commonly used diagnostic tests. A novel assay, the reticulocyte hemoglobin content (CHr), has recently been found to sensitively detect functional iron deficiency among nonuremic patients treated with recombinant erythropoietin (rHuEPO). The purpose of this study was to evaluate the CHr for the assessment of iron status in hemodialysis patients. One hundred sixty-four stable hemodialysis patients had a mean CHr of 27.5 +/- 2.8 pg with a normal distribution of values. The mean CH (mature red cell hemoglobin content) was 26.4 +/- 2.4 pg. There was a close correlation between CHr and CH (r = 0.86, P < 0.0001). A significant subgroup of patients (12.2%) had CHr values < CH. These patients had recent increases in rHuEPO dose, and a lower mean transferrin saturation and hematocrit, suggesting the recent onset of functional iron deficiency due to the increase in rHuEPO dose. In the second phase of the study, 32 patients were randomly selected to receive treatment with a single dose infusion of 1,000 mg of intravenous iron dextran (IVFe). Patients were classified as iron deficient (N = 7) if they responded with a significant reticulocytosis (sustained 1 basis point increase in corrected reticulocyte index within 2 weeks). All other patients were classified as iron replete (N = 25). A CHr < 26 pg at baseline predicted iron deficiency with a sensitivity of 100%, specificity of 80%. The serum ferritin, transferrin saturation and percentage of hypochromic red blood cells all were less accurate. The time to correction of iron deficiency at the level of the reticulocyte was found to be within 48 hours as measured by correction of the mean CHr to > 26 pg, and by the shift of the vast majority of the reticulocyte population to CHr > 26 pg within this time span. We conclude that CHr < 26 pg is an accurate measure of iron status in hemodialysis patients, that a CHr value < CH indicates the acute onset of iron deficiency, and that a single dose infusion of intravenous iron results in correction of iron deficiency at the level of the reticulocyte within 48 hours. PMID- 9211367 TI - Quantitative analysis of ammonia on the breath of patients in end-stage renal failure. PMID- 9211369 TI - Use of urine specific gravity to improve screening for albuminuria. AB - The albumin to creatinine ratio (ACR) can be used to measure urine albumin excretion rates, but is inconvenient and expensive. More rapid and less expensive screening methods estimate only albumin concentration and are subject to errors caused by variation in urine volume. We examined whether urine specific gravity could be used in place of urine creatinine to correct albumin concentration for differences in urine volume in 50 patients. Urine specific gravity accurately estimated urine creatinine concentration (r = 0.79, P < 0.001). The albumin estimated-creatinine ratio (ACestR) in random spot urine sample correlated with urine albumin excretion measured in a 24-hour urine collection (r = 0.98, P < 0.001), as did the ACR (r = 0.95, P < 0.001). For determining microalbuminuria, the sensitivity (0.88) and specificity (0.93) of the ACestR were similar to those of ACR (0.89 and 0.93, respectively). Unfortunately, the sensitivity (0.63) of the Micral-Test was relatively poor, and was only slightly improved by correcting for urine specific gravity (0.69) in this small sample of patients. Nevertheless, these results suggest that as rapid methods for measuring urine albumin concentration improve, combining them with urine specific gravity might produce a less expensive and more convenient alternative to the ACR. PMID- 9211368 TI - A conditionally immortalized cell line from murine proximal tubule. AB - We have developed a conditionally immortalized murine cell line with proximal tubule characteristics (tsMPT) and a background suitable for genetic manipulations. tsMPT was derived from the F1 progeny of crosses between: [1] a transgenic mouse harboring a gamma-interferon (IFN-gamma)-inducible, temperature sensitive SV40 large T antigen gene (tsA58) and [2] mice of the 129/SvEv strain, the background from which most embryonic stem (ES) cells are derived. Under permissive conditions (33 degrees C and in the presence of IFN-gamma), tsMPT cells grow rapidly as monolayers with a doubling time of 23 hours; the large T antigen can be detected by immunocytochemistry and by Western blotting. When transferred to non-permissive conditions (39 degrees C, without IFN-gamma), the cells undergo differentiation coinciding with the disappearance of the large T antigen. By electron microscopy, tsMPT cells are polarized and show microvilli at their apical surface. tsMPT cells express brush border enzymes gamma-glutamyl transpeptidase and carbonic anhydrase IV. They possess Na(+)-dependent transport systems for Pi, D-glucose and L-proline as well as an amiloride-insensitive Na(+) H+ exchanger. Intracellular cAMP generation is stimulated by parathyroid hormone but not by arginine vasopressin. Angiotensinogen mRNA and protein are present in tsMPT with markedly higher levels at non-permissive conditions. tsMPT cells should be a useful model for investigation of the functional features of the proximal tubule epithelium in relation to cellular differentiation. PMID- 9211371 TI - Cyclosporine and the renin-angiotensin axis. PMID- 9211370 TI - A method for accurate measurement of GFR in conscious, spontaneously voiding rats. AB - Renal function measurement by clearance methods relies on accurately timed urine collection. In small experimental animals, renal function measurement is usually performed under anesthesia and/or with the application of bladder catheters to ensure accurate urine collection. To avoid both anesthesia and the need for bladder catheters we developed a method to measure glomerular filtration rate (GFR) in spontaneously voiding conscious rats. GFR was measured as the urinary clearance of constantly infused 125I-iothalamate. To correct for incomplete bladder emptying urinary clearance of 125I-iothalamate was multiplied by the ratio of plasma and urinary clearance of simultaneously infused 131I-hippuran, a correction method that has been previously validated in humans. Reproducibility of the technique was evaluated by analysis of the results of four consecutive clearance periods during the day (intra-assay variation) in a group of 17 rats and of two consecutive clearance periods on two or three separate days in a group of 20 rats (inter-assay variation), all with normal renal function. Application of the correction method reduced the intra-assay coefficient of variation (mean +/- SD) from 37.4 +/- 14.3 to 5.4 +/- 2.3% (P < 0.05). The mean inter-assay coefficient of variation fell slightly from 23.4 +/- 10.3 to 11.0 +/- 7.2% (P < 0.10). In rats with moderately impaired renal function (N = 8) the intra-assay variation fell from 27.9 +/- 20.7 to 2.7 +/- 1.6% (P < 0.05). Our data show that this correction method is a useful technique to assess renal function in conscious, spontaneously voiding rats. PMID- 9211372 TI - Mechanism and echo time dependence of the fast response in fMR. AB - The fast response (FR) is an early reduction in the amplitude of the fMR signal occurring shortly after the onset of stimulation. Owing to its potential advantages in terms of temporal and spatial resolution, it may be of considerable significance in functional experiments. A model for the mechanism of the FR was developed which accounts for the fMRS finding that the amplitude of the dip decreases with increasing echo time. Two computer simulations that confirmed the predictions of the model and showed that it is possible to select optimal echo times for the detection of the FR were performed. PMID- 9211373 TI - VET imaging: magnetic resonance imaging with variable encoding time. AB - A methodology of MRI data acquisition is introduced which involves lengthening the duration of signal readout period with complementary shortening of the phase encode pulses, and vice versa, in sequences where gradient encoding time is limited by RF pulse spacing. This variable encoding time (VET) methodology can be used to increase spatial resolution or reduce data acquisition time in 2D and 3D FT MRI based on CPMG and gradient echo sequences. Advantages of higher image SNR and different spatial frequency distributions in k-space are discussed and evaluated in preliminary experiments. PMID- 9211374 TI - Integrated RF coil with stabilization for fMRI human cortex. AB - Functional brain imaging of the human cortex is limited by poor contrast to noise ratio (CNR) and image degradation due to subject motion during the acquisition period. The work described here combines the use of closely coupled phased array receiver coils with a stabilization system to address these needs. Several phased array designs are evaluated and compared with the conventional "birdcage" design. Coil performance is reported in terms of relative SNR and fMRI results. Relative improvements of up to 360% are obtained for the occipital region and 180% in the temporal region. More modest gains of 10-30% were obtained for a "dome"-shaped birdcage volume coil covering the entire cortex. PMID- 9211375 TI - MRI and NMR spectroscopy of the lipids of atherosclerotic plaque in rabbits and humans. AB - The early stages of atherosclerosis are characterized by the deposition of cholesteryl esters and triglycerides into the arterial wall. In the excised human atherosclerotic plaque these lipids are in a liquid-like state at body temperature and observable via MRI and NMR spectroscopy. To assess the ability of MRI to quantitatively image the lipids of atherosclerotic plaque in vivo, we have investigated eight New Zealand White rabbits fed atherogenic diets (2 weight (wt)% cholesterol, 1 wt% cholesterol + 6 wt% peanut oil, and 1 wt% cholesterol + 6 wt% com oil). Postmortem examination indicated that all rabbits developed atherosclerosis in the aorta. Except for one animal, magnetic resonance angiography showed no noticeable obstruction in the aorta. MRI was carried out in an attempt to image atherosclerotic plaque lipids directly, but no signal was detected in vivo. However, a plaque lipid signal was observed from excised tissue using a small diameter RF coil. 1H NMR spectroscopy of the atherosclerotic plaque from excised aortas indicated that the major fraction of plaque lipids in rabbits is not in a liquid state at physiological temperature and are only marginally MRI visible compared to human plaque lipid. The differences in the MRI characteristics of rabbit and human plaque are due to differences in the fatty acid profile of the cholesteryl esters, chiefly a decrease of linoleic acid in rabbit lesions. PMID- 9211376 TI - Spectroscopic imaging of the water resonance with short repetition time to study tumor response to hyperoxia. AB - A variety of treatments that modulate tumor oxygen tension are used clinically to improve the outcome of radiotherapy. High resolution, noninvasive measurements of the effects of these treatments would greatly facilitate the development of improved therapies and could guide treatment of cancer patients. Previous work demonstrated that magnetic resonance (MR) gradient echo imaging of the water proton resonance detects changes in T2* and T1 in tumors during hyperoxia that may reflect increased tumor oxygenation. This report describes the use of high resolution MR spectroscopic imaging with short repetition time (TR = 0.2 s) to improve the accuracy with which changes in T2* and T1 are measured. Mammary adenocarcinomas grown in the hind limbs of rats were studied. Carbogen inhalation was used to induce hyperoxia. A single 2-mm slice through the center of tumors and underlying muscle was imaged at 4.7 Tesla with in-plane resolution of approximately 1.2 mm and frequency resolution of 5.8 Hz. The peak integral increased by an average of 6% in tumors during carbogen inhalation suggesting a decrease in T1 (n = 8, P < 0.001). Peak height increased by an average of 15% in tumors during carbogen inhalation (n = 8, P < 0.001). The large difference between increases in peak height and peak integral demonstrates that the width of the water resonance decreased. Assuming a Lorentzian lineshape, an average increase of 12% in T2* was observed in tumors. In muscle, peak integral and peak height increased slightly (about 1.2% and 3%, respectively; P < 0.02) during carbogen inhalation but no significant change in T2* was observed. Spectroscopic imaging detects changes in the water proton resonance in tumors during hyperoxia accurately and reproducibly with high signal-to-noise ratio and allows clear separation of T1 and T2* effects. Increases in T2* may be due to decreased deoxyhemoglobin in tumor blood vessels (i.e., the BOLD effect) and may provide a clinically useful index of increases in tumor oxygenation. PMID- 9211377 TI - Dipolar resonance frequency shifts in 1H MR spectra of skeletal muscle: confirmation in rats at 4.7 T in vivo and observation of changes postmortem. AB - Non-isotropic contributions to 1H MR spectra from human skeletal muscle in vivo have recently been observed in the 0- to 5-ppm region. One pair of peaks has been identified to be subject to dipolar couplings. The corresponding changes in resonance frequency are related to the orientation of muscle fibers with respect to the external magnetic field and are analogous to the behavior of small molecules dissolved in liquid crystals. Image-guided localized spectroscopy based on the STEAM method has been applied to verify these phenomena in rat skeletal muscle in vivo and to investigate the effect postmortem. Residual dipolar couplings and anisotropic contributions to 1H MR spectra of skeletal muscle have been confirmed in animals and at a higher field strength--albeit with a slightly different spectral pattern compared to the human study. The most prominent dipolar doublet due to creatine and/or phosphocreatine vanishes postmortem with a rate similar to the disappearance of phosphocreatine, and is no longer observable 2 h postmortem. PMID- 9211378 TI - Skeletal muscle perfusion measurements using adiabatic inversion of arterial water. AB - Quantitative NMR measurements of perfusion using magnetic labeling of arterial water have been demonstrated previously in several different highly perfused organs. The success of these previous experiments suggested that arterial labeling may be of use in measuring perfusion in skeletal muscle, where resting perfusion is very low and where increased perfusion after exercise is transient. In the experiments described in this paper, adiabatic inversion of arterial water has been used to make single-voxel measurements of perfusion in the lower hind limb of rats. At rest, the NMR results were quantified to yield a perfusion rate of about 13.8 ml/100g/min. After perturbation due to ischemic exercise, large relative changes in the NMR signal were observed. The peak change of about 2.5% of the NMR signal occurred shortly after perturbation and was followed by a return to resting levels over a period of about 4 min. PMID- 9211379 TI - Gloxy: an oxygen-sensitive coal for accurate measurement of low oxygen tensions in biological systems. AB - This paper describes the characteristics of a new oxygen sensitive, paramagnetic material that has some significant advantages for measurements of tissue pO2 by in vivo EPR. This paramagnetic component of Welsh coal, termed "gloxy" was found to have valuable EPR features that allow accurate measurement of low oxygen tensions in vivo; these include large oxygen-dependent changes in linewidth, a high number of paramagnetic spin centers (resulting in high signal amplitude), and stability in tissue allowing repeated pO2 measurements to be made in vivo with high precision. Renal pO2 was measured deep in the medulla region of isolated perfused kidneys and found to be lower than that in the cortex (1.7 +/- 0.05 and 7.1 +/- 0.3 mm Hg, respectively). The quality of the EPR signal obtained from the renal outer medulla and also from tumors in mice was such that the pO2 measurements were obtained with a precision of +/-3% of the measured pO2 (Kidney: 1.7 +/- 0.05 mmHg; Tumor: 1.37 +/- 0.04 mmHg). In vitro tests on the viability of cells and in vivo studies using Gloxy demonstrate the stability and inertness of this oxygen-sensitive material. PMID- 9211380 TI - Comparison of blood oxygenation and cerebral blood flow effects in fMRI: estimation of relative oxygen consumption change. AB - The most widely-used functional magnetic resonance imaging (fMRI) technique is based on the blood oxygenation level dependent (BOLD) effect, which requires at least partial uncoupling between cerebral blood flow (CBF) and oxygen consumption changes during increased mental activity. To compare BOLD and CBF effects during tasking, BOLD and flow-sensitive alternating inversion recovery (FAIR) images were acquired during visual stimulation with red goggles at a frequency of 8 Hz in an interleaved fashion. With the FAIR technique, absolute and relative CBF changes were determined. Relative oxygen consumption changes can be estimated using the BOLD and relative CBF changes. In gray matter areas in the visual cortex, absolute and relative CBF changes in humans during photic stimulation were 31 +/- 11 SD ml/100 g tissue/min and 43 +/- 16 SD % (n = 12), respectively, while the relative oxygen consumption change was close to zero. These findings agree extremely well with previous results using positron emission tomography. The BOLD signal change is not linearly correlated with the relative CBF increase across subjects and negatively correlates with the oxygen consumption change. Caution should be exercised when interpreting the BOLD percent change as a quantitative index of the CBF change, especially in inter-subject comparisons. PMID- 9211382 TI - Short TE MR microscopy: accurate measurement and zonal differentiation of normal hyaline cartilage. AB - The purpose of this study was to use MR imaging to accurately measure the thickness of hyaline cartilage and determine the MR contrast parameters for differentiation of cartilage zones in normal human cartilage samples. Cartilage samples were examined using three dimensional spin-echo MR microscopy at 9.4 T with a voxel size of 31 x 31 x 300 microns. Effects of T2 signal loss, susceptibility, and partial volume on measured thickness of cartilage were investigated. Thickness measurements were obtained on corresponding histological sections for comparison. Optimal contrast parameters for delineation of cartilage zones were evaluated using magnetization transfer, inversion recover, T1, and T2 contrast. T2 relaxation losses were identified as the primary source of discrepancy between the measured thickness of cortical bone and hyaline cartilage. Good contrast for zonal differentiation was obtained using T1 weighting. We conclude that images obtained using short TE MR microscopy can be used to accurately measure cartilage and bone thickness in human specimens, and can demonstrate zones within normal cartilage. PMID- 9211381 TI - Dynamics of magnetization in hyperpolarized gas MRI of the lung. AB - The magnetization in hyperpolarized gas (HP) MRI is generated by laser polarization that is independent of the magnet and imaging process. As a consequence, there is no equilibrium magnetization during the image acquisition. The competing processes of gas inflow and depolarization of the spins lead to large changes in signal as one samples k-space. A model is developed of dynamic changes in polarization of hyperpolarized 3He during infusion and in vivo imaging of the lung and verified experimentally in a live guinea pig. Projection encoding is used to measure the view-to-view variation with temporal resolution < 4 ms. Large excitation angles effectively sample the magnetization in the early stages of inflow, highlighting larger airways, while smaller excitation angles produce images of the more distal spaces. The work provides a basis for pulse sequences designed to effectively exploit HP MRI in the lung. PMID- 9211383 TI - Diffusion-weighted multiple shot echo planar imaging of humans without navigation. AB - A new method is proposed for reducing the artifacts produced in diffusion weighted imaging. When data are acquired using multiple shot echo planar acquisitions, conventional reconstruction methods produce artifactual images as a consequence of diffusion weighting and small amounts of bulk motion of the subject. If the amount of motion can be determined, it is possible to correct the data before reconstruction, which removes the artifact. A method for estimating the motion from the acquired data has been developed and evaluated. This method assumes that ghost image effects will be minimized when motion has been correctly compensated. By considering the amount of signal in the background of the image, appropriate corrections to the data can be made, and the accuracy of the motion compensation may be estimated. This technique has been evaluated by computer simulation, and its performance has been demonstrated in a phantom and humans with both two- and four-shot echo planar acquisitions and using both "mosaic" and "interleaved" sampling schemes. PMID- 9211384 TI - Ghost artifact reduction for echo planar imaging using image phase correction. AB - An algorithm is described for reducing ghost artifacts in echo planar imaging (EPI) using phase corrections derived from images reconstructed using only even or odd k-space lines. The N/2 ghost, that arises principally from time-reversal of alternate k-space lines, was significantly reduced by this algorithm without the need for a calibration scan. In images obtained in eight subjects undergoing EPI for auditory functional MRI (fMRI) experiments, N/2 ghost intensity was reduced from 10.3% +/- 2.1% (range: 7.9-14.1%) to 4.5% +/- 0.2% (range: 4.1-4.9%) of parent image intensity, corresponding to a percent reduction in ghost intensity of 54% +/- 9% (range: 43-65%), and the algorithm restored this intensity to the parent image. It provided a significant improvement in image appearance, and increased the correlation coefficients related to neural activation in functional MRI studies. The algorithm provided reduction of artifacts from all polynomial orders of spatial phase errors in both spatial directions. The algorithm did not eliminate N/2 ghost intensity contributed by field inhomogeneities, susceptibility, or chemical shift. PMID- 9211385 TI - Double line scan diffusion imaging. AB - A new double line scan diffusion imaging sequence (DLSDI) is presented. In DLSDI, two lines from two separate slices are acquired in each shot. As its predecessor, LSDI, DLSDI is insensitive to motion artifacts and it can be used on conventional MR scanners. In addition, DLSDI is almost twice as fast as LSDI. Preliminary results from phantom and patient studies show excellent agreement between ADC trace maps obtained with DLSDI and LSDI. The technical and the theoretical aspects of DLSDI are studied, and it is shown how the conditional random walk model can be used as an analytical tool to derive the diffusion sensitivity in the DLSDI sequence. PMID- 9211386 TI - Acceleration mapping by Fourier acceleration-encoding: in vitro study and initial results in the great thoracic vessels. AB - Acceleration mapping can be conducted by replacing the bipolar gradient pulse of a velocity mapping sequence by a tripolar pulse. However, since the acceleration encoding pulse is longer, the image quality is altered by the requirement of a long echo time. Since Fourier encoding velocity imaging has been shown to be robust, this velocity mapping method was transformed into an acceleration mapping method. Four steps of the tripolar acceleration encoding gradient pulse were applied successively; acceleration was then obtained by Fourier transform after zero-filling. The accuracy of the method was assessed with a phantom giving a pulsatile flow. Acceleration maps of the ascending aorta and pulmonary artery were obtained in 10 healthy volunteers. The acceleration values measured were in the range of known physiologic values. The feasibility of Fourier encoding acceleration imaging was also demonstrated in four patients. PMID- 9211387 TI - Density compensation functions for spiral MRI. AB - In interleaved spiral MRI, an object's Fourier transform is sampled along a set of curved trajectories in the spatial frequency domain (k-space). An image of the object is then reconstructed, usually by interpolating the sampled Fourier data onto a Cartesian grid and applying the fast Fourier transform (FFT) algorithm. To obtain accurate results, it is necessary to account for the nonuniform density with which k-space is sampled. An analytic density compensation function (DCF) for spiral MRI, based on the Jacobian determinant for the transformation between Cartesian coordinates and the spiral sampling parameters of time and interleaf rotation angle, is derived in this paper, and the reconstruction accuracy achieved using this function is compared with that obtained using several previously published expressions. Various nonideal conditions, including intersecting trajectories, are considered. The new DCF eliminated intensity cupping that was encountered in images reconstructed with other functions, and significantly reduced the level of artifact observed when unevenly spaced sampling trajectories, such as those achieved with trapezoidal gradient waveforms, were employed. Modified forms of this function were found to provide similar improvements when intersecting trajectories made the spiral-Cartesian transformation noninvertible, and when the shape of the spiral trajectory varied between interleaves. PMID- 9211388 TI - Amplitude demodulation of the electrocardiogram signal (ECG) for respiration monitoring and compensation during MR examinations. AB - A method for respiration monitoring during MR-sequences using the electrocardiogram signal (ECG) is presented. The basic principle is known in conventional patient monitoring and is based on the fact that the amplitude of the ECG-signal varies with respiratory motion. A demodulation of this "cardio respiratory signal" yields a respiration curve that can be used for patient monitoring, triggering, or respiration compensation of MRI sequences. Since no drift is introduced by digital demodulation, this method is superior to analog signal processing. The proposed method was applied during MR examinations using an optically coupled ECG sensor that has the advantage of almost negligible interference due to the MR sequences (< < 10% of the main ECG signal). The respiration signal obtained is strongly correlated (r = 0.79 to 0.98) with the position of the diaphragm in inferior-superior direction measured during breath holding MRI. PMID- 9211389 TI - Efficient design of pulses with trapezoidal magnitude and linear phase response profiles. AB - A variation of the Shinnar-Le Roux (SLR) method of pulse envelope design that allows for control of the phase of the frequency response profile has been developed. The method makes use of the fact that a knowledge of one of the SLR polynomials in combination with a root inversion pattern for the other polynomial is sufficient to fully define the second polynomial. Optimization of the first polynomial, when cast in this form, remains nonlinear. However, it was demonstrated that the relationship between the SLR polynomials and the frequency response profile may be used to generate an initial guess for the SLR polynomials that is sufficiently accurate to allow for the application of linear optimization techniques in most cases. In practice several pulse envelopes having different root inversion patterns are investigated for each target profile. The resulting collection of pulses allows the user to trade off pulse power for profile accuracy. The proposed technique was used to design a large number of amplitude modulated excitation pulses having trapezoidal magnitude and linear phase frequency response profiles. A few examples of the resulting pulses and their response profiles are presented. PMID- 9211390 TI - Stability constants and 1H relaxation effects of ternary complexes formed between Gd-DTPA, Gd-DTPA-BMA, Gd-DOTA, and Gd-EDTA and citrate, phosphate, and carbonate ions. AB - Formation of ternary complexes between Gd-DTPA, Gd-DTPA-BMA, and Gd-DOTA, used as contrast enhancement agents in MRI and the endogenously available carbonate and phosphate ions, has been demonstrated. The extent of ternary complex formation and its effect on the proton relaxation, measured at 9 MHz, rates is negligible at around pH < 8. The complex Gd-EDTA forms more stable ternary complexes with carbonate and phosphate and it also strongly coordinates the terdentate citrate ligand. The formation of ternary complexes Gd-EDTA(X) (X = CO3(2-), Cit3-) results in a significant decrease in the proton relaxation rates under physiological conditions. PMID- 9211391 TI - A spectral approach to analyzing slice selection in planar imaging: optimization for through-plane interpolation. AB - Interpolation between slices is necessary whenever reslicing a volume of data into a different coordinate frame. This may be done to view the data from different perspectives, to align data from different sessions, or to remove the effects of head movement in functional imaging studies. In this paper, issues surrounding slice-selection in two-dimensional imaging are examined in the context of through-plane interpolation and a spectral framework is introduced to describe the sources of error when interpolating between slices. This framework suggests that there is a trade-off between precision in localization, which requires high spatial frequencies, and interpolation, which requires a narrow spectrum of spatial frequencies. An analysis of the sources of error has lead to several approaches to reducing interpolation error including elimination of interslice gaps or making slices overlap, use of a slice profile with a narrower spatial frequency bandwidth such as a Gaussian profile, and use of high-order interpolation. The simulation and experimental data demonstrate significant reductions in interpolation error for these approaches. PMID- 9211392 TI - A comparison of RIGR and SVD dynamic imaging methods. AB - Several constrained imaging methods have recently been proposed for dynamic imaging applications. This paper compares two of these methods: the Reduced encoding Imaging by Generalized-series Reconstruction (RIGR) and Singular Value Decomposition (SVD) methods. RIGR utilizes a priori data for optimal image reconstruction whereas the SVD method seeks to optimize data acquisition. However, this study shows that the existing SVD encoding method tends to bias the data acquisition scheme toward reproducing the known features in the reference image. This characteristic of the SVD encoding method reduces its capability to capture new image features and makes it less suitable than RIGR for dynamic imaging applications. PMID- 9211393 TI - Easy fabrication of a tunable high-pass birdcage resonator. AB - A practical design for a high-pass birdcage resonator is presented. Precision seamless telescoping tubes were used for easy tuning of resonant frequency by adjusting the length of the coils. Three probes, of 4.4, 5.0, and 25.0 cm in diameter, respectively, were constructed and tested. An empirical formula is given that can be used to calculate the capacitance needed for a given frequency when the desired physical dimension and the number of elements of the coll are specified. A simple three-step procedure is suggested for easy fabrication of resonators that are routinely tunable over tens of megahertz. PMID- 9211394 TI - Alternate k-space sampling in EPI: possible sources of ringing artifacts. PMID- 9211395 TI - A systems approach to molecular structure, intermolecular recognition, and emergence-dissolvence in medicinal research. PMID- 9211396 TI - Chemoprotection: a review of the potential therapeutic antioxidant properties of green tea (Camellia sinensis) and certain of its constituents. PMID- 9211397 TI - Recent advances in the discovery and development of topoisomerase inhibitors as antitumor agents. PMID- 9211398 TI - Blood flow structuring and its alterations in capillaries of the cerebral cortex. AB - Various manifestations of blood flow structuring were investigated in rabbit cerebral cortex capillaries, which possess the most narrow lumina of all parts of the body. Blood flow structuring in the capillaries was characterized by the presence of a stable and comparatively large parietal plasma layer, which changed insignificantly under control and ischemic conditions, but disappeared when blood stasis developed inside the capillaries. The axial core of the blood flow in the capillaries, which occupied almost two-thirds of the intracapillary volume under normal conditions, consisted of significantly deformed (stretched along the microvessels' axes) and nonaggregated erythrocytes. During ischemia the shape of the erythrocytes did not change appreciably; only the blood plasma intervals between them increased significantly, demonstrating reduction of the local hematocrit. During primary blood stasis caused by enhanced intravascular erythrocyte aggregation, typical blood flow structuring became significantly disturbed: red cells filled the whole, or almost the whole, capillary lumina and did not leave visible space for plasma inside the microvessel lumina. We concluded that normal blood flow structuring is a deciding factor in the blood rheological properties of microvessels. Its disturbance, caused by fast accumulation of erythrocytes in the capillary lumina, results in blood rheological disorders and in a slow down to a full stop of the blood flow, despite a preserved arteriolovenular pressure difference. PMID- 9211399 TI - Retinal artery and vein pressures in the dog and their relationship to aortic, intraocular, and cerebrospinal fluid pressures. AB - The relationship between retinal arterial (Pra) and aortic (Pa) pressures is unknown, and the relationship between retinal vein (Prv) pressure and intraocular pressure (IOP) is not clear. Also unclear is the effect of cerebrospinal fluid pressure (CSFp) upon retinal venous pressure. We aimed to measure the relationships among Pra, Prv, Pa, IOP, and CSFp. Dogs were anesthetized while IOP, CSFp, and Pa were monitored. Pipettes with 2.5-micron diameter tips, connected to a servonulling pressure transducer, were used to record pressures from the retinal arteries and veins. Across a range of IOP (16-22 mmHg), CSFp (0 21 mmHg), and Pa (23-195 mmHg) the Pra = 0.72 Pa + 4.3 (r = 0.99, n = 61, P < 0.01), which suggests that the relationship between Pra and Pa is linear over a broad range of systemic blood pressures. The correlation coefficient between Prv and IOP was greater than 0.96 (P < 0.01) at all venous sites and whether IOP was greater than or less than CSFp. The transmural pressure varied along the retinal vein from 1.3 +/- 0.3 mmHg (+/-95% CI, n = 30) at 1 disk diameter from the optic disk rim to 0.3 +/- 0.2 mmHg (n = 66) at the optic disk, with a 0.9-mmHg/mm pressure gradient. These are the first measurements demonstrating a retinal vein transmural pressure close to zero. PMID- 9211400 TI - The skin blood flow response in wound healing. AB - Although vasodilation is conventionally held to be the predominant microvascular response to a wound, there has been no previous attempt to actually quantitate skin blood flow within and in the neighborhood of wounds. In particular, there has been no differentiation between sites with primarily nutritive (NUTR) blood flow and those with considerable arteriovenous (AV) perfusion. We used our previously described model of cutaneous blood flow in the rat to study the blood flow response to wounding. We measured skin blood flow at the centers and at the undisturbed perimeters of wounds placed at the back, a NUTR site, and at the paw, an AV site, in 11 Wistar Kyoto rats. Measurements were performed at baseline, and then at 3 hr, 24 hr, 72 hr, and 7 days postwounding. At 3 hr, flow at the center of the back wound had increased to 11.3 +/- 1.4 ml/min/100 g from a baseline of 2.1 +/- 0.1 ml/min/100 g and remained elevated at 7 days (8.3 ml/min/100 g). Flow at the perimeter of the back wound rose as well, but not as high as at wound center, to twice the baseline level (4.1 ml/min/ 100 g at Day 7). Flow values at control sites on the back did not increase from baseline. Flow at the center of the paw wound rose from 7.2 +/- 0.5 ml/min/100 g at baseline to 15.6 +/- 4.3 ml/min/100 g at Day 3 but then fell back to 6.9 +/- 0.9 ml/min/100 g at Day 7. There was only a very small increase in the basal temperature wound response at the paw perimeter. Blood flow at all wound sites showed a response to heat. At the back, heating to 44 degrees stimulated an 80% increase in blood flow at baseline. This degree of increase was maintained at both the center and the perimeter of the back wound. In contrast, although there was also a thermal response at the paw wound center, it was of much lower magnitude than the nonwounded baseline response. As a result, the heat-stimulated flow value actually fell over the 7 days to approximately half of the baseline level. At the paw wound periphery, there was an initial fall in the heat stimulated response, but it then recovered to the baseline level and remained stable over the 7 days. Thus, the skin blood flow response seen at the paw wound challenges the conventional concept of vasodilation as the expected wound blood flow response. The mechanisms of blood flow response in the healing wound may be more complex than the simple inflammatory vasodilation conventionally postulated. PMID- 9211401 TI - Capillary adrenoceptors in rat skeletal muscle. AB - The purpose of this study was to examine whether functional alpha- and beta adrenoceptors exist on capillaries of rat skeletal muscle, and further to determine which subtype of these receptors predominates on these capillaries. Using intravital video microscopy, we measured red blood cell velocity (VRBC) responses in capillaries of rat extensor digitorum longus muscle (EDL) following a local application of these agonists: norepinephrine (NE; alpha 1, alpha 2; 10( 7) to 3 x 10(-3) M), phenylephrine (PE; alpha 1; 3 x 10(-4) to 10(-2) M), clonidine (CLO; alpha 2; 3 x 10(-3) to 10(-2) M), UK14304 (alpha 2; 3 x 10(-4) to 10(-2) M), and isoproterenol (IPR; beta 1, beta 2; 10(-7) to 3 x 10(-3) M). Responses to NE (10(-5) M) were also measured after a local pretreatment with prazosin (alpha 1 antagonist; 10(-5) to 10(-3) M) and rauwolscine (alpha 2 antagonist; 3 x 10(-4) to 3 x 10(-2) M), while responses to IPR (10(-5) M) were measured after local atenolol (ATE; beta 1 antagonist; 10(-3) to 10(-2) M) and butoxamine (BUT; beta 2 antagonist; 10(-3) to 10(-2) M) pretreatment. The overall control VRBC was 226 microns/sec. NE, PE, CLO, and UK14304 resulted in concentration-dependent decreases of VRBC (from -12 to -89%) from the control level, while IPR caused concentration-dependent increases (17 to 174%). PE reduced VRBC to a larger degree than CLO and UK14304. NE-induced VRBC responses tended to be attenuated more by prazosin than by rauwolscine. Both ATE (10(-2) M) and BUT (10(-3) and 10(-2) M) alone decreased VRBC. However, only ATE significantly attenuated the IPR-induced VRBC responses. These results suggest that the capillary of rat EDL muscle has alpha- and beta-adrenoceptors. From the two alpha-adrenoceptor subtypes, the capillary may be predominated by the alpha 1 adrenoceptors. PMID- 9211402 TI - Evidence for K+ channels involvement in capillary sensing and for bidirectionality in capillary communication. AB - Although the capillary sensing and communication phenomenon has been characterized, its mechanism is not clear. It has been hypothesized that capillary sensing involves a membrane potential change in the capillary endothelium and/or pericyte and that communication represents an electrotonic spread of this change along the capillary. The goal of the present study was to address this hypothesis by examining the presence of K+ channels on the capillary and by determining bidirectionality of communication. Using intravital microscopy, we locally applied K+ (100 mM), acetylcholine (ACh; 3 mM), and norepinephrine (NE; 0.3 mM) on capillaries, 400-500 microns downstream from the arteriole, at the surface of the sartorius muscle in anesthetized frogs. Responses were measured in terms of red blood cell velocity (VRBC) changes in the stimulated capillary (control prestimulation VRBC ranged from 110 to 770 microns/sec). K+ and ACh caused significant 19 and 38% increases in VRBC, while NE caused a -46% decrease, respectively. The K+ response was blocked by local pretreatment with K+ channel blocker BaCl2 (1 microM) and by pretreatment with tetraethyl ammonium chloride (TEA; 5 mM). Responses to ACh and NE were attenuated by pretreatment with 1 microM BaCl2 (to 1%) and with 50 mM TEA (to -25%), respectively. In a separate experiment, NE (3 mM) application on the capillary 500 microns away from the draining venule (capillary occluded) caused a 19% venular constriction (i.e., similar to a reported 21% arteriolar constriction caused by the NE stimulus). We concluded that (i) K+ channels were present on the capillary and (ii) capillary communication was bidirectional. We interpreted these results to be consistent with the above hypothesis of membrane potential change and electrotonic spread. PMID- 9211403 TI - Urokinase-type plasminogen activator overexpression enhances the invasive capacity of endothelial cells. AB - Fetal bovine aortic endothelial GM 7373 cells were transfected with a viral expression vector harboring the human urokinase-type plasminogen activator (uPA) gene. The stable transfectant clone uPA-R5 overexpressed and secreted human uPA as shown by Northern blot analysis, immunoprecipitation of metabolically labeled proteins, plasmin chromogenic assays, and SDS-PAGE zymography of cell extracts and conditioned media. The uPA-R5 cells were analyzed for their invasive capacity in vitro in the Matrigel chemoinvasion assay in the presence of serine- or metalloprotease inhibitors. uPA overexpression enhanced the invasive capacity of GM 7373 cells through a mechanism which differs from that mediated by metalloproteases. Endothelial cell uPA transfectants may represent an useful experimental model to investigate the role of uPA in angiogenesis and angioproliferative diseases. PMID- 9211404 TI - Isometric tension of cultured endothelial cells: new technical aspects. AB - In this paper new technical aspects are discussed in the measurement of the low amount of force typically expressed in cultured endothelial cells. We illustrate how potential background noises interfere with signal acquisition. We present a new generation prototype that measures isometric tension in vitro in multiple samples and in more than on isometric vector. We report that thrombin increases isometric tension in at least two separate vectors that are directed in opposite directions. We also report that phorbol ester dibutyrate can randomly mediate a false relaxation (anisotropic contraction) in cultured PPAEC, when the force vector is directed opposite to the referenced isometric vector of the transducer. In contrast, stimulation of cultured HUVEC with the cAMP agonists, theophylline and forskolin, decreased isometric force in both vectors. Thus direction of the force vector needs to be considered when interpreting isometric tension in cultured endothelial cells. PMID- 9211405 TI - A novel instrument for studying the flow behaviour of erythrocytes through microchannels simulating human blood capillaries. AB - A novel instrument has been developed to study the microrheology of erythrocytes as they flow through channels of dimensions similar to human blood capillaries. The channels are produced in silicon substrates using microengineering technology. Accurately defined, physiological driving pressures and temperatures are employed whilst precise, real-time image processing allows individual cells to be monitored continuously during their transit. The instrument characterises each cell in a sample of ca. 1000 in terms of its volume and flow velocity profile during its transit through a channel. The unique representation of the data in volume/velocity space provides new insight into the microrheological behaviour of blood. The image processing and subsequent data analysis enable the system to reject anomalous events such as multiple cell transits, thereby ensuring integrity of the resulting data. By employing an array of microfluidic flow channels we can integrate a number of different but precise and highly reproducible channel sizes and geometries within one array, thereby allowing multiple, concurrent isobaric measurements on one sample. As an illustration of the performance of the system, volume/velocity data sets recorded in a microfluidic device incorporating multiple channels of 100 microns length and individual widths ranging between 3.0 and 4.0 microns are presented. PMID- 9211406 TI - Immunolocalization of endothelin and nitric oxide-synthase in lymphatic vessels and cultured lymphatic endothelial cells. PMID- 9211407 TI - Vaccine for atherosclerosis. PMID- 9211408 TI - Ferrous iron chelator to treat subarachnoid haemorrhage. PMID- 9211409 TI - Alpha-particle-emitting radioisotopes coupled to antibody for acute myeloid leukaemia treatment. PMID- 9211410 TI - Hepatocyte growth factor, tissue repair and cancer. PMID- 9211411 TI - Molecular medicine's applications to the developing world: enlightened self interest. PMID- 9211413 TI - Cancer genetics. PMID- 9211412 TI - Papillomavirus: biology and clinical implications for immunotherapy. PMID- 9211414 TI - Prevention of mother-to-infant transmission of HIV-1. AB - The number of children with AIDS continues to increase worldwide. Children who become infected with HIV-1 acquire the infection almost exclusively from their mothers during pregnancy or delivery, or via breast feeding. Mother-to-infant transmission has been, and continues to be, an area of active research with the goal being complete prevention. Treatment with zidovudine, an antiviral agent, has been found to decrease transmission from 25% to 8%. However, multiple obstacles impede the worldwide application of this advance. PMID- 9211415 TI - The SCID-hu mouse: an in vivo model for HIV-1 infection in humans. AB - The lack of suitable animal models for the in vivo study of HIV-1 infection has prompted investigators to take advantage of the graft-rejection deficit in severe combined immunodeficient (SCID) mice. Two separate approaches have been used to transplant human lymphoid and/or hemolymphoid tissues into SCID mice to generate chimeric animals in which distinct elements of the human immune system could be maintained and studied in vivo. The two models that arose from this work were the SCID-hu mouse and hu-PBL-SCID mouse. The goal of producing these distinct models is to provide an easily manipulable model for the in vivo analysis of HIV-1 infection and its ensuing pathophysiology. Both models support HIV-1 replication and display potential as models for studying antiviral strategies and mechanisms of viral pathogenesis. This review focuses on the SCID-hu mouse. PMID- 9211416 TI - Biological properties of dendritic cells: implications to their use in the treatment of cancer. AB - Dendritic cells are the principal initiators of antigen-specific immune responses. The mechanisms by which they activate resting, naive T cells are increasingly well understood. Dendritic cells have several molecules on their surface that are critical for T-cell activation; they secrete multiple soluble factors that are important for T-cell differentiation and growth; and they home to areas in lymphoid organs that are rich in effector T cells. This knowledge has led to the belief that delivery of antigens with dendritic cells that have been manipulated ex vivo could stimulate powerful cellular immune responses against tumors. Pilot clinical trials indicate that dendritic cell vaccines can induce efficacious tumor-specific immune responses. PMID- 9211417 TI - Human tumour antigens recognized by T cells: new perspectives for anti-cancer vaccines? AB - Cytolytic T-lymphocytes (CTLs) mediate tumour rejection in several animal models. In humans, presuming that T cells might be able to eradicate cancer cells as effectively as they eliminate virus-infected cells, an exciting challenge for tumour immunologists is (1) to identify specific CTL-targeted antigens on these cancer cells and (2) to manipulate these antigens so that they can initiate or amplify the patient's native immune response, which would otherwise be insufficient. This review focuses on the identification of several tumour antigens, their molecular nature, and how they can be used to develop anti-cancer vaccines. PMID- 9211418 TI - Central nervous system actions of oxytocin and modulation of behavior in humans. AB - The posterior pituitary hormone oxytocin has modulatory effects on neural functioning that are significant to the regulation of behavior. Basic research in animals has established the importance of oxytocin in affiliation, including mating, pair bonding and parenting behaviors. It is also an important regulator of feeding, grooming and responses to stress. The actions of oxytocin in the brain are regulated by gonadal steroid hormones, particularly estrogen. Oxytocin might also influence normal behavior in humans, and dysfunctions in the oxytocin system might be involved in the etiology and expression of neuropsychiatric disorders. PMID- 9211420 TI - Equine inhibin/activin beta A-subunit mRNA is expressed in the endometrial gland, but not in the trophoblast, during pregnancy. AB - The expression of both inhibin alpha- and inhibin/activin beta A-subunit mRNA was examined in equine uteroplacental tissues collected during pregnancy (days 90 to 300). Northern blot analysis revealed that 5 transcripts (7.0, 4.1, 3.4, 2.6, 1.5 kb) of beta A-subunit were present, and the most abundantly expressed transcript was the 1.5 kb one. Relatively high levels of the 1.5 kb transcript were seen in the second trimester of pregnancy compared to what was found in the third trimester. To identify the tissue localization of beta A-subunit mRNA, in situ hybridization was performed, and the positive signal was observed exclusively in the endometrial glands, but not in the fetal placental tissue (trophoblast) at days 150, 210, and 300 of pregnancy. On the other hand, inhibin alpha-subunit transcript could not be detected at any stage of pregnancy examined either by Northern blot analysis or in situ hybridization. Although the factor(s) regulating the gene expression of beta A-subunit in this equine tissue is currently unknown, these results suggest that activin, but not inhibin, is predominantly produced in the endometrial glands of the pregnant mare, and thus produced activin may play a paracrine or endocrine role during pregnancy in this species. PMID- 9211419 TI - Identification of maternal transcripts that progressively disappear during the cleavage period of rabbit embryos. AB - In order to characterize the changes that occur in the population of maternally inherited transcripts before the transition from maternal to zygotic control of embryonic development (MZT) in mammals, we used rabbit embryos where zygotic transcription becomes necessary only after the fourth cleavage division, during the second day that follows fertilization. In the present work we have associated mRNA differential display and an RT-PCR based-method that allows amplification of the whole population of messengers to identify and characterize maternal transcripts which are degraded throughout this early period of development. While there is no major degradation of the polyA RNA population before MZT we identify 4 transcripts which progressively disappear up until the 8-16 cell stage. We also show that the degradation of one of these maternal messengers is controlled by zygotic transcription, which is not the case for the three others. This messenger shows homology with the human FGF9 gene and is potentially a good candidate to address the question of the molecular control of maternal to zygotic transition in early mammalian embryogenesis. PMID- 9211421 TI - Molecular cloning and tissue-specific expression of the mouse homologue of the rat brain 14-3-3 theta protein: characterization of its cellular and developmental pattern of expression in the male germ line. AB - The highly conserved 14-3-3 family of proteins, originally reported as brain specific and then found in various somatic cells and oocytes, interacts with several important signal transduction kinases so that actually the 14-3-3 protein are considered as modulators of multiple signal transduction pathways. Here we show that a 14-3-3 protein is also expressed in the male germ cells, thus extending the protein cellular distribution to a cell line never reported to express 14-3-3 proteins. Screening of a mouse spermatogenic cells lambda gt11 cDNA library with affinity-purified polyclonal antibodies to the tyrosine kinase SP42 allowed the isolation of several positive clones. Sequencing of a positive cDNA clone revealed a 735-nucleotide open reading frame encoding a protein of 245 amino acids (27,778 Da). The predicted protein was found to be identical to the most recently discovered 14-3-3 isoform, the theta subtype from a rat brain. Here we demonstrate that 14-3-3 theta mRNA is highly expressed in testis and brain only. Western immunoblot analyses confirm the Northern blot data. Developmental Northern and Western blot analyses are consistent with an expression and translation of the 14-3-3 theta gene throughout spermatogenesis. However, analysis of RNA from purified populations of spermatogenic cells at different developmental stages and immunohistochemistry on adult testis sections reveal that within the testis the 14-3-3 theta gene products are most abundant in meiotic prophase spermatocytes, and, above all, in differentiating spermatids. Both testicular and epididymal spermatozoa are negative. The present study is the first report on the presence and molecular characterization of the 14-3-3 theta gene product in the male germ line. Our observations suggest that this specific member of the 14-3-3 protein family could play distinct modulatory roles in the complex development of the mammalian male germ cell lineage. PMID- 9211422 TI - Up-regulation and down-regulation of genes expressed in cocultures of rat Sertoli cells and germ cells. AB - To better understand the molecular interactions between somatic and germ cells in the mammalian testis, we have begun to analyze with mRNA differential display changes in gene expression induced by coculturing rat Sertoli cells and germ cells. We have identified 10 cDNAs that are either down-regulated or up-regulated in cocultures of germ cells and Sertoli cells. Three genes expressed in Sertoli cells and three genes expressed in germ cells were down-regulated in Sertoli cell germ cell cocultures, whereas four genes were up-regulated in the cocultures. Northern blot analysis was used to establish the expression pattern of the mRNAs encoded by the cDNAs and to define the sizes of the differentially expressed mRNAs. Sequence analysis of the cDNAs and computer searches against the GenBank and EMBL DNA databases were used to relate the ten cDNAs to known genes. Of the three Sertoli cell cDNAs, one appeared identical to transferin, while the other two shared regions of similarity to an endoplasmic reticulum stress protein and to a pro-alpha 2 XI collagen, respectively. The three germ cell cDNAs shared sequences with fibronectin, with a basic fibroblast growth factor receptor and with an IgG gamma 2b, respectively. The four cDNAs that were up-regulated in the Sertoli-germ cell cocultures showed similarity to an isoform of casein kinase 1 delta, to an epidermal growth factor, to a statin-related protein, and to an integral membrane glycoprotein. These data demonstrate that a number of specific genes are up- and down-regulated when germ cells and Sertoli cells are cocultured, and suggest these genes are important in cell to cell communication during spermatogenesis. PMID- 9211424 TI - Activator effect of coinjected enhancers on the muscle-specific expression of promoters in zebrafish embryos. AB - The transient expression of reporter gene constructs in embryos provides a powerful tool to characterise cis-acting transcriptional elements of the genes involved in development. In the present study, we have analysed the expression pattern of several muscle-specific and ubiquitous regulatory sequences in microinjected zebrafish embryos. By using a fast and reproducible coinjection strategy, the mosaic expression of lacZ reporter gene was monitored in wholemount embryos injected with sequences containing putative enhancer elements and a carp myosin heavy chain promoter/lacZ reporter construct. We have found that a 0.9-kb myosin heavy chain (MyHC) proximal promoter containing several putative myogenic regulatory factors (MRF) binding sites is sufficient to restrict lacZ expression to the skeletal muscle fibres of prim-6 stage zebrafish embryos. Expression of a rat-derived foetal myosin light chain enhancer (MyLC) and different fragments of a carp beta-actin regulatory region together with the MyHC promoter were compared by accumulating the type, number and spatial distribution of beta-galactosidase expressing cells on an expression map. beta-galactosidase activity increased similarly whether the MyLC enhancer was ligated to the promoter/ reporter construct directly or when coinjected as a separate fragment whilst skeletal muscle specificity was retained. The coinjection of two different forms of the beta-actin regulatory elements also showed a marked effect on the MyHC promoter activity. The coinjection of putative enhancers with minimal promoter constructs and subsequent analysis of the transient expression pattern in the developing embryos provides a rapid and simple technique to identify cis acting activator elements of genes expressed in the vertebrate embryo. PMID- 9211425 TI - Changes in the relative abundance of various housekeeping gene transcripts in in vitro-produced early bovine embryos. AB - The relative abundances of transcripts of different origins and housekeeping functions were measured by Northern blot analysis of RNA samples derived from in vitro-matured oocytes and in vitro-produced bovine embryos at selected stages of early development. The gene products studied included: two mitochondrial transcripts, 12S rRNA and cytochrome b mRNA; two RNAs involved in the processing of other RNAs, U2 and U3 snRNA; and two nuclear-derived transcripts, beta-actin mRNA and histone H3 mRNA. Overall, the RNA levels for the various genes studied remained constant or decreased slightly from the mature oocyte to the 6- to 8 cell or morula stage and were greatly increased in blastocysts. Differences were observed in the degree to which the RNA levels increased and in the timing of the increase. For 12S rRNA, a major increase was not observed until the blastocyst stage where levels increased 7.1 times the amount detected in morulae. Cytochrome b mRNA levels started to increase at the 6- to 8-cell stage and reached levels in blastocysts that were 20 times more than the cytochrome b mRNA level in 2- to 4 cell embryos. U2 snRNA levels did not increase until the blastocyst stage where levels were 6.4 times the amount found in morulae. U3 snRNA and beta-actin mRNA levels started to increase at the morula stage and blastocysts contained 118 and 110 times more U3 snRNA and beta-actin mRNA, respectively, than 6- to 8-cell embryos. However, blastocysts contained only two times the amount of histone H3 mRNA present in 6- to 8-cell embryos. PMID- 9211423 TI - Repression of MHC class II gene transcription in trophoblast cells by novel single-stranded DNA binding proteins. AB - The maintenance of the fetus during pregnancy has been attributed to the absence of major histocompatibility complex (MHC) class II antigens on fetal trophoblastic cells that make contact with the maternal immune system. However, the mechanism(s) by which class II genes are regulated in trophoblast cells is unclear. We have identified a negative regulatory element (IA alpha NRE) in the promoter of the mouse class II gene IA alpha that represses IA alpha transcription in trophoblast cells. IA alpha NRE, located from-839 to -828, binds transacting factors from rat, mouse and human trophoblast cells, but not from 18 other cell lines tested. These results indicate that IA alpha NRE binding proteins (IA alpha NRE BPs) are conserved in species with hemochordial placentas, and suggest that IA alpha NRE binding activity is restricted primarily to trophoblast cells. Interestingly, the IA alpha NRE BPs bind to the IA alpha NRE antisense strand in a sequence-specific manner. IA alpha NRE represses transcription from the IA alpha promoter in a position-dependent manner, and has a minor down-regulatory effect on the activity of the SV40 promoter/enhancer. Our results demonstrate that MHC class II gene transcription is repressed in fetal trophoblast cells by sequence-specific, single-stranded DNA binding proteins, and suggest a possible mechanism by which the conceptus is protected from immune rejection during pregnancy. PMID- 9211426 TI - Stage-dependent redistributions of acetylated histones in nuclei of the early preimplantation mouse embryo. AB - In the preimplantation mouse embryo, activation of the embryonic genome is accompanied by a transient enrichment of histone H4 acetylated at lysines 5, 8, and 12 at the nuclear periphery (Worrad et al., 1995: Development 121:2949-2959). In the present report, we use laser-scanning confocal microscopy and a new panel of antibodies to define the distribution of specific acetylated isoforms of the other three core histones in mouse embryos at the 1- to 4-cell stage. We find that histone H3 acetylated at lysine 9 and/or 18 (H3.Ac9/18) and the single acetylated form of H2A (H2A.Ac5) become transiently enriched at the nuclear periphery in the 2-cell embryo. In contrast, H3.Ac14, H3.Ac23, and acetylated H2B, like H4.Ac16, remain distributed throughout the nucleoplasm. The staining intensity with antisera to H3.Ac9/18, even at the periphery was weak compared to that obtained with antisera to acetylated H4. A brief period of culture, however, in the presence of the inhibitor of histone deacetylases trichostatin A (TSA) or trapoxin increased labeling. Thus, the steady-state level of H3.Ac9/18 at the nuclear periphery and H3.Ac14 and H3.Ac23 in the nuclear interior is relatively low, but turnover remains high. The localization of selected acetylated isoforms of H3 and H2A at the nuclear periphery was independent of ongoing transcription or of cytokinesis, but did require DNA replication. We propose a model in which the selective, replication-dependent acetylation and deacetylation of zygotic chromatin at the nuclear periphery mediates the programming of zygotic transcription. PMID- 9211427 TI - Role of the first round of DNA replication in reprogramming gene expression in the preimplantation mouse embryo. AB - The first round of DNA replication has been proposed to provide a window of opportunity for maternally-derived transcription factors to gain access to their cognate cis-binding DNA sequences and thereby reprogram the pattern of gene expression that occurs during the 2-cell stage. Using high-resolution, two dimensional electrophoresis of metabolically radio-labeled polypeptides, we report the expression of a group of several polypeptides whose synthesis in the 2 cell embryo is due to transcription (i.e., alpha-amanitin-sensitive) and depends on the first round of DNA replication (i.e., aphidicolin-sensitive). We also describe the synthesis of another subset of alpha-amanitin-sensitive polypeptides whose expression does not require the first round of DNA replication (i.e., aphidicolin-insensitive). These results are consistent with the proposed role of the first round of DNA replication in reprogramming the pattern of gene expression. PMID- 9211428 TI - Effects of low temperatures on in vitro produced bovine zygotes. AB - The objective of this research was to investigate the effects of cooling on the development of bovine zygotes. One-cell bovine embryos were maintained at 39 degrees C (control), 20 degrees C, 10 degrees C, or 0 degree C for 5, 10, or 20 minutes, then cultured in vitro for 7 days and the proportion of embryos developing to the compact morula or blastocyst stage compared between different treatments. Duration of exposure time had no effect on development. Development rates to the compact morula or blastocyst stage were 3.9%, 11.4%, 17.4%, and 24.4% for zygotes maintained at 0 degree C, 10 degrees C, 20 degrees C, and 39 degrees C, respectively, with differences in embryo yield between every treatment (P < 0.05). In a second experiment, bovine pronuclei (karyoplasts) and cytoplasts were cooled at 0 degree C or maintained at 39 degrees C for 5 minutes. Pronuclear transplantation was then utilized to create 4 types of reconstructed embryos, those with: 1) non-cooled pronuclei and non-cooled cytoplasm, 2) non-cooled pronuclei and cooled cytoplasm, 3) cooled pronuclei and non-cooled cytoplasm, and 4) cooled pronuclei and cooled cytoplasm. The proportion of embryos developing to the blastocyst stage was highest when non-cooled pronuclei were transferred into non-cooled cytoplasm (18.9%), and similar to that of non-cooled, non-manipulated control zygotes (13.2%, P > 0.05). No embryos developed to the blastocyst stage when pronuclei (cooled or non-cooled) were transferred into cooled cytoplasm. However, zygotes with cooled pronuclei transferred into non-cooled cytoplasm yielded 4.5% blastocysts (P < 0.05). More embryos developed to the compact morula or blastocyst stage when non-cooled vs. cooled cytoplasm was utilized, regardless of whether the pronuclei were cooled (P < 0.05). These data demonstrate that pronuclei are more tolerant to low temperature exposure than is ovum cytoplasm. PMID- 9211429 TI - Succinate and malate improve development of hamster eight-cell embryos in vitro: confirmation of viability by embryo transfer. AB - The in vitro development of hamster preimplantation embryos is supported by non glucose energy substrates. To investigate the importance of embryonic metabolism, influence of succinate and malate on the development of hamster 8-cell embryos to blastocysts was examined using a chemically defined protein-free modified hamster embryo culture medium-2 (HECM-2m). There was a dose-dependent influence of succinate on blastocyst development; 0.5 mM succinate was optimal (85.1% +/- 3.9 vs. 54.5% +/- 3.5). In succinate-supplemented HECM-2m, blastocyst development was reduced by omission of lactate (68.5% +/- 7.2), but not pyruvate (85.8% +/- 6.2) or glutamine (84.1% +/- 2.1). Succinate along with either glutamine or lactate or pyruvate poorly supported blastocyst development (28%-58%). Malate also stimulated blastocyst development; 0.01 mM malate was optimal (86.3% +/- 2.8). Supplementation of both succinate and malate to HECM-2m supported maximal (100%) blastocyst development, which was inhibited 4-fold by the addition of glucose/phosphate. The mean cell numbers (MCN) of blastocysts cultured in succinate-supplemented HECM-2m was higher (28.3 +/- 1.1) than it was for those cultured in the absence of glutamine or pyruvate (range 20-24). The MCN was the highest (33.4 +/- 1.6) for blastocysts cultured in succinate-malate-supplemented HECM-2m followed by those in succinate (28.3 +/- 1.1) or malate (24.7 +/- 0.5) supplemented HECM-2m. Embryo transfer experiments showed that 29.8% (+/- 4.5) of transferred blastocysts cultured in succinate-malate-supplemented HECM-2m produced live births, similar (P > 0.1) to the control transfers of freshly recovered 8-cells (33.5% +/- 2.0) or blastocysts (28.9% +/- 3.0). These data show that supplementation of succinate and malate to HECM-2m supports 100% development of hamster 8-cell embryos to high quality viable blastocysts and that non-glucose oxidizable energy substrates are the most preferred components in hamster embryo culture medium. PMID- 9211430 TI - Estrogen and progesterone receptors and estrogen receptor-related antigen (ER-D5) in human epididymis. AB - Estrogen receptor mRNA expression was detected in the head and tail parts of the human epididymis by means of reverse transcription polymerase chain reaction (RT PCR). Immunocytochemical labelling showed that ciliate and nonciliate cells from the epithelium of the efferent ductules possessed strong to moderate nuclear staining for estrogen and progesterone receptors. The epididymal duct was negative for both antigens. Vascular endothelial cells also showed estrogen receptor, but no progesterone receptor immunolabelling in all regions of the organ. Immunoreactivity for the estrogen receptor-related antigen (ER-D5) was seen in the cytoplasm of ciliated and nonciliated epithelial cells from the efferent ductules, in the basal cells of some epididymal duct profiles as well as in myofibroblasts of the lamina propria, endothelial cells and smooth muscle cells of the vessel walls. The results obtained suggest that the epithelial cells of the efferent ductules of the human epididymis may be main target structures of estrogens and progestins. PMID- 9211431 TI - Timing of meiotic progression in bovine oocytes and its effect on early embryo development. AB - This study was designed to investigate the effect of the kinetics of nuclear maturation in bovine oocytes on early embryo development and to examine whether the time of insemination of mature oocytes affects the oocytes' ability to support events of early embryo development. The time required for completion of nuclear maturation was influenced by gonadotropins used to supplement the maturation medium. Luteinizing hormone (LH) enhanced the speed of nuclear maturation when compared to follicle-stimulating hormone (FSH). Oocytes completing their nuclear maturation early (by 16 hours after the initiation of culture) were more likely to complete the first embryonic cell cycle (78% in LH vs. 43% in FSH) and develop to the blastocyst stage (47% in LH vs. 34% in FSH). As the age of the oocytes at the time of MII arrest increased (extrusion of the polar body by 20 or 24 hours), a decrease in their ability to cleave and develop to the blastocyst stage was observed. Differences in the oocyte's ability to decondense chromatin and form pronuclei were also observed. Early maturing oocytes started forming pronuclei earlier than their later maturing counterparts. The time of insemination of mature oocytes played an equally important role. Generally, when insemination of mature oocytes was delayed for 8 hours, higher proportions of fertilized oocytes developed to advanced preimplantation stages than did the oocytes inseminated immediately after metaphase II arrest. PMID- 9211432 TI - Reactive oxygen species generation by human spermatozoa is induced by exogenous NADPH and inhibited by the flavoprotein inhibitors diphenylene iodonium and quinacrine. AB - Human spermatozoa possess a specialized capacity to generate reactive oxygen species (ROS) that is thought to be of significance in the redox regulation of sperm capacitation (De Lamirande and Gagnon, 1993; Aitken et al., 1995). However, the mechanisms by which ROS are generated by these cells are not understood. In this study we have examined the possible significance of NADPH as a substrate for ROS production by human spermatozoa. Addition of NADPH to viable populations of motile spermatozoa induced a sudden dose-dependent increase in the rate of superoxide generation via mechanisms that could not be disrupted by inhibitors of the mitochondrial electron transport chain (antimycin A, rotenone, carbonyl cyanide m-chlorophenylhydrazone [CCCP], and sodium azide), diaphorase (dicoumarol) xanthine oxidase (allopurinol), or lactic acid dehydrogenase (sodium oxamate). However, NADPH-induced ROS generation could be stimulated by permeabilization and was negatively correlated with sperm function. Both NADH and NADPH were active electron donors in this system, while NAD+ and NADP+ exhibited little activity. Stereo-specificity was evident in the response in that only the beta-isomer of NADPH supported superoxide production. The involvement of a flavoprotein in the electron transfer process was indicated by the high sensitivity of the oxidase to inhibition by diphenylene iodonium and quinacrine. These results indicate that NAD(P)H can serve as an electron donor for superoxide generation by human spermatozoa and present a simple strategy for the production of motile populations of free radical generating cells with which to study the significance of these molecules in the control of normal and pathological sperm function. PMID- 9211433 TI - Binding of rat and ovine epididymis-specific prealbumins (PES) to rat spermatozoa without effect of heterologous immunization on rat fertility. AB - The epididymis, under control of testosterone, secretes proteins which bind to the membrane of the spermatozoa during their passage through the lumen. One such class is termed PES (prealbumin epididymal specific). Injection of heterologous oPES (ovine PES) into male rats caused antibody production but failed to induce sterility, unlike results previously obtained when rat PES was injected into male rats. This suggests that only very restricted species-specific epitopes of PES might be useful for causing immunocontraception. Despite this, the sperm binding properties of PES purified from the rat (rat PES) and from the ram (oPES) were shown to be similar. When either rat PES or oPES, conjugated with a fluorescent probe (dimethylamino-fluorescein), was incubated with washed rat spermatozoa originating from the caput, corpus or cauda epididymis, results of flow cytometric analysis showed: (1) the number of spermatozoa bound to isologous or heterologous fluorescent PES, and (2) the binding-affinity of spermatozoa for PES was greater for sperm collected from more distal sites in the epididymis. PMID- 9211434 TI - Direct evidence for the secretion of lactoferrin and its binding to sperm in the porcine epididymis. AB - Lactoferrin has been for the first time purified from the porcine cauda epididymal fluid as a 70 kDa protein. Both Western and Northern blot analyses show that lactoferrin is synthesized in the regions from the distal caput to the cauda epididymis and secreted into the luminal fluid. Lactoferrin is first secreted as a 75 kDa glycoprotein and its carbohydrate moieties are gradually digested to form 70 kDa protein in the cauda epididymis. Lactoferrin has already bound to the surface of the epididymal sperm because the anti-lactoferrin antiserum induces the mature sperm tail-to-tail agglutination. These results strongly suggest new physiological functions of lactoferrin on the sperm maturation in the epididymis. PMID- 9211435 TI - Alteration in dopaminergic function in abstinent alcoholics. AB - In order to evaluate the tuberoinfundibular endogenous dopaminergic tone in alcoholic subjects, the inhibitory effect of an infusion of dopamine (3 micrograms/kg/min for 2 h) on PRL secretion was tested in 11 alcoholics after 4 weeks of abstinence. On different days alcoholics were tested with TRH to evaluate possible alterations in the PRL pituitary reserve. Age-matched normal men participated as controls. In addition, the status of cerebral structures, such as the frontal-subcortical area, where dopamine plays an important role as neurotransmitter, was evaluated in all subjects by radiological (CT scan) and functional (neuropsychological tests) studies. The PRL response to TRH was similar in the two groups. In contrast, dopamine-induced PRL decrement was significantly lighter in alcoholics than in controls. Neuroradiological and neuropsychological parameters were abnormal in alcoholics. These data suggest an alteration in dopaminergic activity involving the tuberoinfundibular and probably the fronto-subcortical system in 4 weeks abstinent alcoholics. PMID- 9211436 TI - Human leukocyte antigen system in clozapine-induced agranulocytosis. AB - Forty-three schizophrenic patients participating in this study were serotyped for human leukocyte antigens (HLA-A, -B, -C, -DR, -DQ antigens). Thirty-six of them were hospitalised in two state mental hospitals and 7 in our general hospital, psychiatric unit. The patients from our unit were typed for HLA before commencing clozapine treatment whereas the patients from state hospitals were typed after commencing treatment. Three out of 43 patients developed agranulocytosis. One had a combination of both 'high-risk' haplotypes (HLA-B16(38,39), DR4, DQ3 and HLA DR2, DQ1), another had HLA-DR2, DQ1, whereas the last had a totally different haplotype. Between non-agranulocytic patients 1 was found to carry the HLA B16(38,39), DR4, DQ3 haplotype and 14 (out of 40) had the HLA-DR2, DQ1. Taking into account other factors supposed to be involved (a noxious metabolite, and the presence of a humoral cytotoxic factor) we must admit that despite the finding of a high-risk haplotype in Jewish populations there are other aspects of this question awaiting clarification. PMID- 9211437 TI - Cortisol response to d-fenfluramine in patients with obsessive-compulsive disorder and in healthy subjects: evidence for a gender-related effect. AB - In order to evaluate serotonergic function in obsessive-compulsive disorder (OCD), plasma cortisol response to d-fenfluramine (30 mg p.o.) was examined in 20 drug-free obsessive-compulsive patients (10 males and 10 females) and in 20 age- and sex-matched healthy subjects, under double-blind, placebo-controlled conditions. We found that: (a) baseline plasma cortisol secretion was significantly increased in patients with OCD; (b) in healthy subjects, the cortisol response to d-fenfluramine was evident in women, but no in men; (c) plasma cortisol response to the serotonergic challenge did not differ between patients and controls, but it was significantly reduced in female patients as compared to healthy women. These results demonstrate a hyperactivity of the hypothalamo-pituitary-adrenal axis in obsessive-compulsive patients and suggest a dysfunction of 5-HT transmission in female patients. PMID- 9211438 TI - A preliminary report of a strong genetic component for thought disorder in normals. A twin study. AB - Several authors have investigated the presence of thought disorder in psychiatric patients using different reliable methods. Under the hypothesis of a genetic predisposition to thought disorder, the degree and quality of thought disorder have also been studied in populations at a high risk for psychosis, in particular for schizophrenia. As a result, an increasing incidence of thought disorder was detected in relatives of schizophrenics. To account for the thought disorder also found in normal subjects, researchers propose that thought disorder exists in normal subjects on a continuum with schizophrenic patients. In the following report, we evaluated the inherited component of thought disorder in normal subjects, using a sample of 25 normal twin pairs, 16 monozygotic and 9 dizygotic twin pairs. We applied the Thought Disorder Index (TDI) to assess disordered thinking, genetic estimates were made with classical methods, controlling for environmental sources of variability where possible. Our findings suggest a strong additive genetic component for the global TDI rating variable, with a heritability estimate approaching 80-90%. New approaches in neuropsychology and neuropsychiatry-based on genetic methodologies should further define the cerebral physiology responsible for disordered thinking. PMID- 9211440 TI - Medicated chronic schizophrenic patients do not demonstrate left turning asymmetry. AB - Twenty-seven medicated chronic schizophrenic female patients were tested for right or left turning behavior. No substantial right or left asymmetry was found, nor did the addition of the indirect dopaminergic agonist amantadine influence these results. Previous studies demonstrated left circling preference in chronic unmedicated schizophrenics and it seems that this previous finding is abolished by neuroleptic treatment. However, our patients were all females and further research with a heterogenous group of schizophrenic patients is needed. PMID- 9211439 TI - Platelet 5-HT levels and hypothalamic-pituitary-adrenal axis activity in schizophrenic patients with positive and negative symptoms. AB - Hypothalamic-pituitary-adrenal (HPA) axis activity and platelet 5-HT concentrations were determined before dexamethasone suppression test (DST) in 80 male schizophrenic patients with predominantly positive or negative symptoms. Significant differences in platelet 5-HT and no differences in baseline plasma cortisol concentrations among schizophrenic suppressors and nonsuppressors were found. A similar rate of nonsuppression (56% positive and 53% negative schizophrenic patients) was detected. Platelet 5-HT, but not plasma cortisol concentrations, could be used to differentiate positive and negative symptoms of schizophrenia. PMID- 9211441 TI - The treatment of panic disorder in a clinical setting: a 12-month naturalistic study. AB - In the treatment of panic disorder double-blind controlled studies have demonstrated that imipramine (IMI) was effective at doses higher than 125 mg/day. However a high rate of dropouts because of side effects has been reported in these subjects. In clinical settings, the administration of benzodiazepines (BDZ) in combination with IMI has been proposed to reduce the frequency and severity of side effects. In this naturalistic study, 49 patients affected by panic disorder with agoraphobia (n = 36) or without agoraphobia (n = 13) were treated with IMI plus lorazepam (LRZ) and followed for 12 months. The mean effective doses were 63.5 +/- 35.5 mg/day for IMI and 2.4 +/- 1.3 mg/day for LRZ. During the follow-up period, panic attacks disappeared in 75.5% of patients and 69.5% of agoraphobics were free of phobic avoidance. The patients with comorbid mood disorders and longer duration of illness required higher doses of both drugs. The combined treatment of IMI and LRZ allowed the use of low doses of the drugs, reduced the frequency and severity of the side effects and improved patient compliance. In fact, only 1 patient (2%) dropped out because of the severity of side effects. Furthermore, the patients who tapered LRZ treatment did not show BDZ withdrawal syndrome. PMID- 9211442 TI - Plasma alpha-one acid glycoprotein and haloperidol concentrations in schizophrenic patients. AB - Thirty six schizophrenic patients were randomly assigned to placebo or haloperidol treatment for 6 weeks. Blood samples to measure plasma alpha-one acid glycoprotein (AAG), haloperidol and reduced haloperidol concentrations were obtained at baseline and weeks 2, 4, and 6. Blood samples were obtained 10-12 h after the evening dose and prior to the morning dose. Haloperidol and reduced haloperidol was assayed by HPLC with electrochemical detection. Plasma AAG levels were assayed by radial immunodiffusion. Patients were clinically assessed by the Brief Psychiatric Rating Scale (BPRS) and Abnormal Involuntary Movement Scale at baseline and weeks 2, 4, and 6. BPRS scores did not significantly decrease during placebo treatment, although a slight drop in plasma AAG levels was found. Haloperidol produced a significant decrease in BPRS scores and plasma AAG levels. Mean plasma haloperidol levels were 12.9 +/- 14.7 ng/ml at week 6. Significant correlations between decreasing BPRS scores and plasma AAG levels were not found with only a strong trend at week 2 (r = 0.445, p = 0.073). The role of AAG and psychotropic drug disposition in psychiatric patients requires further evaluation. PMID- 9211443 TI - Value of thyroid echography in the long-term follow-up of lithium-treated patients. AB - Psychiatric patients on long-term lithium (Li) therapy frequently develop goiter and/or hypothyroidism. It has also been suggested that Li may trigger/exacerbate thyroid autoimmunity. Previous studies provided evidence that underlying thyroid diseases represent important predisposing factors for the development of Li induced thyroid dysfunction. The aim of the present paper was to assess the value of thyroid ultrasound-a simple and reliable tool to detect subtle thyroid abnormalities-in the longitudinal evaluation of 23 Li-treated psychiatric patients without evidence of biochemical thyroid abnormalities before therapy. For this purpose, thyroid ultrasound was associated with a clinical and laboratory (serum thyroxine, serum triiodothyronine, serum TSH, antithyroglobulin (AbTg), antithyroid microsomal (AbM) and antithyroid peroxidase autoantibodies) evaluation prior to and at 6- to 12-month intervals during Li treatment. On the basis of thyroid ultrasound before Li, patients were subdivided into two groups: group A (n = 15, 7 males, 8 females) with a normal echography and group B (n = 8, 5 males, 3 females) with mild ultrasound abnormalities. In group A the development of a small diffuse goiter was confirmed by physical examination during Li therapy; 2 patients displayed a transient increase of serum TSH concentration and none developed detectable serum antithyroid autoantibodies. Beside the small volumetric increase, no other ultrasound abnormalities were observed during the entire follow-up. In all group B patients a mild diffuse goiter was clinically detected before and on Li administration and no significant volumetric changes were observed during follow-up. Two patients developed high titers of AbM and AbTg 12 and 18 months after the beginning of Li, respectively; in 1 a persistent increase of serum TSH concentration was also observed. Thyroid echography before Li displayed different degrees of scattered or diffuse hypoechogenicity and a further decrease in echogenicity was detected during Li therapy in 2 patients. In conclusion, we provided further evidence that long-term Li administration is not associated with de novo appearance of thyroid autoimmune phenomena in humans, but rather with an exacerbation of underlying thyroid autoimmunity. In addition to thyroid autoantibody and TSH measurements, thyroid echography appears to be a sensitive tool in the identification of patients at risk of developing autoimmune hypothyroidism during long-term Li therapy. PMID- 9211444 TI - Effects of the tranquillizer diazepam and the stimulant methylphenidate on alertness and memory. AB - Effects of alertness and memory of a single dose of diazepam (10 mg) and the central stimulant methylphenidate (20 mg) were studied in healthy volunteers. It was questioned whether opposite effects of diazepam and methylphenidate are not only observed with respect to alertness but also with respect to memory. It was also questioned whether the two drugs equally affect the first (primacy) and last (recency) items in both the immediate and delayed recall of newly learned words. The experiment was performed in a double-blind, placebo-controlled way. 12 subjects were exposed to a subjective alertness scale and a verbal memory test: a 15-word test. Subjective alertness was found to be decreased after diazepam and increased after methylphenidate. Anterograde amnesia was found after diazepam in the memory test. More specifically, the primacy but not the recency effect was reduced during the immediate recall and both were reduced in the delayed recall. methylphenidate had no effect on memory, however a ceiling effect might have obscured a putative drug effect. The results of a second experiment excluded this possibility. In all, the data demonstrate opposite effects of the two drugs on subjective alertness, suggesting opposite effects on vigilance. Opposite effects on memory were not established. This demonstrates that changes in alertness do not run in parallel with changes in memory. A scatter diagram, however, suggests a small effect of alertness on immediate recall. The effects of diazepam were also discussed in terms of the Atkinson and Shiffrin memory theory and it seems that diminished rehearsal processes are one of the key factors in explaining diazepam-induced amnesia. PMID- 9211445 TI - Baseline electroencephalographic tracings in rabbits differ significantly according to 'acute' or 'chronic' preparation. A computerized study. AB - Baseline electroencephalographic (EEG) tracings recorded from 'acute' and 'chronic' rabbits were compared by computerized spectral analysis. The results showed that baseline EEG activity of rabbits differ significantly and markedly according to acute or chronic preparation. It is suggested that this finding should be taken into account when studying the EEG effects of centrally acting drugs. PMID- 9211446 TI - Event-related potential changes in cocaine withdrawal: evidence for long-term cognitive effects. AB - The effects of cocaine withdrawal on the latency of the P300 cognitive event related potential were studied in 36 subjects. All subjects had used cocaine within 48 h of admission to an inpatient unit. P300 was recorded in a traditional auditory oddball paradigm 2 and 6 days following admission. P300 latency decreased from day 2 to day 6 in subjects with a history of intravenous use, but was unchanged in subjects who had only used cocaine orally. The neurotransmitter systems mediating this effect, and the differential cognitive effects of route of administration are discussed. PMID- 9211447 TI - [Ca2+ channels in the central nervous system]. AB - Roles of Ca2+ channels in physiological functions of mammalian central synapses were discussed from a system-oriented point of view. In the presynaptic terminals of the mammalian CNS so far studied, synaptic transmission is mediated by the subclass of Ca2+ channels designated as the N-type (alpha 1B channels) and/or by that designated as the P/Q-type (alpha 1A channels). In some central synapses such as those between neocortical pyramidal neurons, synaptic transmission is presynaptically suppressed by various transmitter-modulators. Our electrophysiological data indicate that the receptors for amines, glutamate, GABA and adenosine co-exist on individual terminals, and they exert a common modulatory effect on synaptic transmission. Details of the intracellular cascade, i.e., G-protein and Ca2+ channel subtypes that are linked in this modulation, remain to be elucidated. Although the direct 'membrane delimited' action of G proteins on Ca2+ channels is strongly suggested as a modulatory mechanism by the resemblance to the modulation observed in other neurons, the indirect second messenger pathways, however, may also be involved in the control of Ca2+ channels. Postsynaptically located Ca2+ channels are considered to play important roles in the regulation of neuronal excitability and synaptic plasticity. Individual dendritic spines apparently serve as a primary unit in an increase in Ca2+ level. This compartmentalized increase of Ca2+ seems essential for determining plastic changes of the synaptic efficacy in those particular spines. There is ample evidence indicating that the postsynaptic Ca2+ channels are involved in this Ca2+ transient. In order to understand the physiological significance of Ca2+ channels in CNS functions, further elucidation of channel subtypes, intracellular cascades of the modulator actions and characterization of the channel modifications will be essential. PMID- 9211448 TI - [Role of the renin-angiotensin system in cardiovascular and renal diseases]. AB - Accumulating evidence indicate that various growth-related genes, growth factors and extracellular matrix components play a central role in the pathogenesis of cardiovascular and renal diseases by regulating cellular phenotype, growth and migration or promoting tissue fibrosis. Treatment of hypertensive rats with an angiotensin II type 1-receptor (AT1-receptor) antagonist normalizes cardiac phenotypic modulation and the increased fibrosis-related gene expressions in hypertrophied heart, leading to the improvement of cardiac dysfunction. The AT1 receptor antagonist can inhibit protooncogenes (c-fos, c-jun and Egr-1) and fibronectin gene expressions in rat balloon-injured artery, which is associated with the inhibition of neointima formation. Furthermore, the AT1-receptor antagonist prevents either the phenotypic modulation of glomerular mesangial cells or the increase in transforming growth factor-beta 1 expression in nephrosclerosis. Thus, the AT1-receptor antagonist in vivo potently inhibits the expression of growth-related gene and extracellular matrix and inhibits cellular phenotypic modulation. The AT1-receptor is responsible for the pathogenesis and development of cardiovascular and renal diseases. PMID- 9211449 TI - [A sensitive two-site (sandwich) enzyme immunoassay system for measuring nerve growth factor (NGF)]. AB - Nerve growth factor (NGF) is a prototype of neurotrophin family neurotrophic factors that have nearly 50% amino acid sequence homology with each other. NGF is known to support survival and stimulate differentiation of sympathetic and neural crest-derived some sensory neurons in the peripheral nervous system and the basal forebrain cholinergic neurons in the central nervous system. Recent investigations have expanded NGF action to much wider cell populations than previously known. Namely, NGF stimulates differentiation and/or proliferation of immune cells including lymphocytes, leucocytes and mast cells. These findings suggest important roles of NGF in the crosstalk between the nervous system and immune system. Therefore, a practical and sensitive method to quantify physiological NGF is in great demand. We developed a sensitive two-site enzyme immunoassay (EIA) system for mouse NGF, based on the sandwiching of the antigen between anti-mouse NGF antibody IgG coated to a plastic plate and biotinylated anti-NGF antibody. Avidin-beta-D-galactosidase was then bound to biotines, and galactosidase activity was determined fluorometrically. This method is simple, rapid and sensitive. Purified NGF is detectable at a concentration as low as a few pg/ml, which is enough to detect physiological NGF in various tissues and NGF secreted from cultured non-neuronal cells such as fibroblasts, astrocytes and Schwann cells. PMID- 9211450 TI - Readability of breast self-exam literature. PMID- 9211451 TI - Identification and education of persons with a hereditary predisposition to malignancy. PMID- 9211452 TI - Pediatric dermatoses: three common skin disruptions in infancy. AB - Skin disruptions account for 20% to 30% of pediatric primary care visits [1]. These disruptions may result from skin infections, inflammatory responses, insect bites, and infestations. This article focuses on the identification and management of skin disruptions related to inflammatory dermatoses. The most common dermatoses in infancy are seborrheic dermatitis, (also known as cradle cap); diaper or primary contact dermatitis; and atopic dermatitis, more commonly referred to as eczema, an entity that has yet to be clearly defined. Recognition and appropriate treatment of these common pediatric dermatoses must not just focus on the skin disruptions; it is important that the infant be assessed within the context of the family. The primary care provider must be aware that these conditions have the potential to affect the developing relationship between the infant, parent(s), and family. The practitioner within the provider-family relationship, through education and support, can empower the parent(s) to provide the necessary care for their infant. PMID- 9211453 TI - A comprehensive and efficient process for counseling patients desiring sterilization. AB - To optimize the time spent counseling a sterilization patient, this article presents a 10-step process that includes all steps necessary to ensure a comprehensive counseling session: (1) Discuss current contraception use and all available methods; (2) assess the client's interest in/readiness for sterilization; (3) emphasize that the procedure is meant to be permanent, but there is a possibility of failure; (4) explain the surgical procedure using visuals, and include a discussion of benefits and risks; (5) explain privately to the client the need to use condoms if engaging in risky sexual activity; (6) have the client read and sign an informed consent form; (7) schedule an appointment for the procedure and provide the patient with a copy of all necessary paperwork; (8) discuss cost and payment method; (9) provide written preoperative and postoperative instructions; and (10) schedule a postoperation visit, or a postoperation semen analysis. PMID- 9211454 TI - Hyperthyroidism: diagnosis and management of Graves' disease. AB - Hyperthyroidism, or thyrotoxicosis, results when the body's tissues are exposed to excessive levels of thyroid hormone. Hyperthyroidism affects 2% of women but only one-tenth as many men. Graves' disease is the most common form of hyperthyroidism, often occurring in young adults. It is an autoimmune disorder with an important genetic component. Hyperthyroidism's hallmarks include goiter and myriad signs and symptoms related to increased metabolic activity in virtually all body tissues. Increased sensitivity to circulating catecholamines adds to the clinical picture. Diagnosed by patient history, physical examination, and laboratory tests, Graves' disease is treated with antithyroid drugs, radioactive iodine, and/or surgery, plus supportive therapy. A good treatment outcome can be expected; long-term follow-up is indicated. PMID- 9211455 TI - Drug-nutrient interactions in enteral feeding: a primary care focus. AB - Drug and nutrient interactions are complex and can take many forms, including malabsorption of either the drug or the nutrient component. Some drugs can stimulate or suppress appetite, whereas others can cause nausea and vomiting resulting in inadequate nutritional intake. Absorption of drugs is a complex process that can be affected by the physical characteristics of the gastrointestinal tract (GIT) as well. Depending on the physical properties of a drug, it may be absorbed in a limited area of the GIT or more diffusely along much of the entire length. Many diseases and conditions are also known to affect the GIT either directly or indirectly. Dietary factors also need to be considered when the "food" is an enteral formula. The widespread use of enteral tubes requires that consideration be given to patients receiving both enteral feedings and medication concurrently. The location of a tube in the gastrointestinal tract, as well as the problems involved in crushing and administering solid dosage forms, creates a unique set of problems. PMID- 9211457 TI - Post polio syndrome: an update for the primary health care provider. AB - Post Polio Syndrome, or PPS, is defined as a clinical syndrome of new weakness, fatigue, and pain in people who have previously recovered from acute paralytic poliomyelitis. Other common symptoms include cold intolerance, dysphagia, dyspnea, and overuse syndromes. PPS afflicts an estimated 50% of polio survivors, a population estimated at 1.6 million people, and begins roughly 30 years after the acute disease. The main impact of PPS is on mobility related activities affecting one's daily routine. With an insidious onset, and several differential diagnoses for each symptom, PPS can be difficult to diagnose and to validate. However, once identified, there are treatment plans and many avenues of support for this disabling syndrome. The purpose of this article is to provide an overview of the pathophysiology of both acute paralytic poliomyelitis as well as PPS. This article also reviews the current literature concerning the etiology and pathophysiology of both poliomyelitis and PPS, symptom evaluation and differential diagnoses, and treatment recommendations. The psychosocial impact and care of the client are also identified, and several resources for support and education of both the client and provider are provided. PMID- 9211456 TI - Detecting, diagnosing, and preventing oral cancer. AB - Unlike many other malignancies, cancers of the mouth and surrounding tissues continue to cause considerable mortality and morbidity in this country. Therefore, early detection, diagnosis, and treatment of patients with oral cancer must be a high priority for all health care providers. This review is aimed at heightening the awareness of clinicians to the early signs and symptoms of oral cancer. Recognition of early lesions is crucial for improved long-term patient survival. Factors such as advanced age, tobacco and/or alcohol use, chronic sum exposure, and a previous diagnosis of cancer can alert clinicians to patients who may be at risk for developing oral cancer. Because most oral malignancies are asymptomatic and may mimic benign conditions, any suspicious lesion should be carefully examined and, if appropriate, referred immediately for histological examination. Measures such as annual oral cancer screening examinations and patient education that stress early signs and symptoms of oral cancer can also help to reduce the risk in high-risk individuals. PMID- 9211459 TI - Regional or multistate licensure: is it coming soon? PMID- 9211458 TI - The emergence of acyclovir resistance in mucocutaneous herpes simplex viral infections: implications for clinical practice. PMID- 9211460 TI - Professional behaviors in nurse practitioners. PMID- 9211461 TI - Gender bias in health care. PMID- 9211462 TI - Using nurse practitioner certification for state nursing regulation: an update. PMID- 9211463 TI - A new chromatographic method for measurement of rubidium transport activities in cultured bovine retinal pigment epithelial cells. AB - A new method for measuring cellular rubidium (Rb+) uptake activities based on cation chromatography was developed and compared with the standard technique, uptake of the radioisotope 86Rb+, using cultured bovine retinal pigment epithelial (RPE) cells. The Rb+ response was strictly linear from 0.25 nmol (detection limit) to 25 nmol. The Na+/K(+)-ATPase inhibitor ouabain inhibited Rb+ uptake with IC50 values of 128.7 +/- 23.5 nM (n = 8; radioactive method) and 56.6 +/- 9.3 nM (n = 9; non-radioactive method, p < 0.01). The latter value is identical to the IC50 value of 54.4 +/- 16.2 nM (n = 3) for ouabain binding to the intact RPE cells. Ouabain and bumetanide, an inhibitor of the Na+/K+/Cl(-) cotransporter, each inhibited Rb+ uptake maximally by 66 and 30%, respectively. This new technique allows sensitive measurement of intracellular Rb+, as well as K+ and Na+, and, thus, should prove useful for studying the effects of pharmacologic agents and simulated disease conditions on cation transport and cation balance in RPE and other cell types. PMID- 9211464 TI - Monocyte-macrophage differentiation induced by coculture of retinal pigment epithelium cells with monocytes. AB - Macrophages play an important role in proliferative vitreoretinopathy (PVR). Since macrophage maturation may be modulated by the local microenvironment, we determined the effect of retinal pigment epithelium (RPE) cells interacting with monocytes on macrophage maturation. The enriched monocyte fraction of peripheral blood mononuclear cells was cocultured with RPE cells. Cell-free supernatants conditioned during the culture periods 0-4 days (growing RPE cells) and 4-8 days (confluent RPE cells) were tested for their capacity to induce monocyte/macrophage differentiation of the promyelocytic cell line HL-60, which was measured by the expression of CD11c and CD14 and flow cytometry. RPE cells released factors that increased the CD14 expression on HL-60 cells in terms of percentage of positive cells (22.7% vs. control 10.4%). RPE-cell-conditioned supernatants had no effect on the CD11c expression. Monocytes secreted substances that increased the expression of CD11c (20.5% vs. control 9.1%; p = 0.003) and CD14 (31.6% vs. control 10.4%; p < 0.0001). Supernatants from cocultures increased the CD11c (19.8%) and CD14 expression (40.8%) to values that were similar to the sum of those of cell monocultures. Supernatants conditioned during the later culture had no effect on CD14 and CD11c expression. We conclude that invading monocytes and RPE cells could create an intraocular microenvironment that supports macrophage maturation during the initial stage of PVR. PMID- 9211465 TI - Vascular expression of endothelial antigen PAL-E indicates absence of blood ocular barriers in the normal eye. AB - The endothelium-specific antigen PAL-E is expressed in capillaries and veins throughout the body with the exception of the brain, where the antigen is absent from anatomical sites with a patent blood-brain barrier. In this study we determined vascular endothelial staining for PAL-E in the normal eye in relation to the ocular blood-tissue barriers. Immunohistochemical staining of frozen tissue sections of eyes from 22 cornea donors and a number of normal animal autopsy eyes was performed for the PAL-E antigen and the blood-brain barrier marker glucose transporter 1. In normal human and animal eyes, endothelial PAL-E staining was absent from the microvasculature in iris, ciliary muscle, optic nerve and retina. In a few normal human eyes, some weakly stained capillaries were observed in the retina and nerve fiber layer, mostly in the peripapillary area. Marked staining of capillaries and venules with PAL-E was observed in the conjunctiva, episclera, sclera, ciliary processes, choriocapillaris and optic nerve head. In general, the endothelial antigen PAL-E is absent from microvessels involved in the blood-ocular and the blood-retinal barriers. PAL-E may therefore be a useful marker to identify pathological breakdown of blood-ocular barriers. PMID- 9211466 TI - Effect of timolol and UF-021 (a prostaglandin-related compound) on pulsatile ocular blood flow in normal volunteers. AB - The effects of topically applied timolol (a nonselective beta-blocker) and UF-021 (a prostaglandin-related compound) on pulsatile ocular blood flow (POBF) were investigated in 9 healthy volunteers. One drop of either UF-021 or timolol was instilled in one randomly selected eye and the fellow eye received physiologic saline. Before and 90 min after drop instillation, we measured intraocular pressure (IOP), POBF, heart rate (HR) and blood pressure (BP). After a washout interval of at least 1 week, the other drug was instilled in the previously treated eye. IOP, POBF, HR, and PB were measured following the same protocol. After timolol administration, IOP and POBF decreased significantly in the treated eye (26%, p = 0.01, and 13%, p = 0.02, respectively) and in the fellow eye (11%, p = 0.01, and 11%, p = 0.03, respectively). HR also decreased (10%, p = 0.03), but BP did not change significantly. After UF-021 administration, no significant changes were found in IOP, POBF, HR, or BP. Our results suggest that in normal subjects timolol decreases POBF in both the treated and the untreated eyes, whereas UF-021 has no action on either IOP or POBF. PMID- 9211468 TI - Histochemical study of leukocyte elastase activity in alkali-burned rabbit cornea. AB - In order to contribute to a better understanding of the role of leukocyte elastase in corneal melting after a severe alkali burn, the in situ appearance and activity of this enzyme in the rabbit cornea was examined. For this reason, a histochemical approach using cryostat sections on membranes and a gel incubation medium with the very sensitive substrate Mu-Ala-Ala-Pro-Val-7-amino-4 trifluoromethylcoumarin (Enzyme Systems Products, Livermore, Calif., USA) was employed. In contrast to the normal rabbit cornea where leukocyte elastase activity was absent, in alkali-burned cornea a high enzyme activity in inflammatory cells (particularly polymorphonuclear leukocytes) was present. The extracellular release of leukocyte elastase into the substantia propria of the corneal stroma was associated with disintegration of the corneal stroma and the appearance of corneal ulcers. These results show that leukocyte elastase is associated with corneal destruction after a severe alkali burn. PMID- 9211467 TI - Combined non-steroidal therapy in experimental corneal injury. AB - PURPOSE: The effects of anti-inflammatory non-steroidal therapy combined with free-radical scavengers were studied and compared to corticosteroid use in the treatment of experimental corneal injury. METHOD: Eighty New Zealand albino rabbits were used in this study. A corneal alkali burn was induced by applying 1 N NaOH filter paper on the central axis of the right cornea for 30 s. Animals were distributed into five treatment groups: group 1 (control group) was only given gentamicin; group 2 was treated with 0.5% dimethylthiourea (DMU); group 3 received 1% dexamethasone; group 4 was given combined 0.5% DMU and 1% indomethacin; group 5 was treated with 0.5% DMU and 0.1% diclofenac sodium. One 50-microliter drop of gentamicin was instilled every 12 h, whereas the other drugs were instilled every 6 h (50 microliters). All groups received the same antibiotic treatment as the control group. The animals were killed on the 5th day. Inflammatory index, area and perimeter of the wounded corneal zone, and corneal transparency were evaluated. RESULTS: No significant differences in the inflammatory index were found between the treatment groups and the control group after 72 h. Significant differences (p < 0.001) were observed at 24 h in groups 3 5 when compared with the control group. Planimetry showed significant differences in group 4 when compared with the other groups (p < 0.05). Corneal transparency study showed statistically significantly better values in groups 4 and 5, when compared with the other groups, including group 3 (p < 0.05). CONCLUSIONS: The use of 0.5% DMU combined with 1% indomethacin can be considered an alternative to corticosteroid treatment in our experimental chemical corneal injury. PMID- 9211469 TI - The Cryner element in the murine gamma-crystallin promoters interacts with lens proteins. AB - Based upon DNA sequence analysis of the promoters from six gamma-crystallin genes (cryga-->crygf) a 36-bp DNA fragment was defined as 'Cryner' (cryg nested repeat). The presence of these repeats made this structure a candidate for DNA protein interaction. The present experiments demonstrate interactions of lens proteins with the Cryner element from murine cryga, crygb, crygd and cryge. Additionally, DNA covering the sequence of about 30 nt between Cryner and the TATA-box of the murine crygb exhibits sequence-specific interactions with the bovine alpha-crystallin-containing fraction. The results confirm the hypothesis that the Cryner element is able to interact with lens proteins. It is noteworthy that this interaction is specific for the template strand of the DNA. The present model includes the possibility of sequence-dependent conformational changes leading to various DNA-protein complexes. PMID- 9211470 TI - Polycyclic aromatic hydrocarbons in bovine lens. AB - A study was performed to detect the presence of polycyclic aromatic hydrocarbons (PAHs) in bovine eyes. Twenty fresh bovine eyes were used to detect the presence of PAHs in lens, vitreous and aqueous by HPLC and spectrofluorometer. In lenticular tissue the mean amount of PAHs was 0.0271 microgram/g and the mean level of PAH in each lens was 0.059 microgram. Five types of PAHs (pyrene, fluoranthene, triphenylene, 1.2-benzanthracene and chrysene) were found in the lenses but none in vitreous and aqueous. These data indicate that PAHs are present only in the lens of the bovine eye. The source of these substances in mammalian clear lens is unclear. PMID- 9211471 TI - Peripheral morphine analgesia. PMID- 9211472 TI - Life-threatening adverse reactions after acupuncture? A systematic review. PMID- 9211473 TI - Pain relief from intra-articular morphine after knee surgery: a qualitative systematic review. AB - Reduction of postoperative pain by injecting opioid into the knee joint is believed to support the hypothesis of peripheral opioid receptor activation in inflammation. The study design consisted of a systematic review of randomised controlled trials (RCTs). Main outcomes were pain intensity and the use of supplementary analgesics. Efficacy of intra-articular bupivacaine against placebo was used as an index of internal sensitivity. Evidence of efficacy was sought in both early (0-6 h after intra-articular injection) and late (6-24 h) periods. Thirty-six RCTs in knee surgery were found. Six had both a local anaesthetic control and placebo; four showed internal sensitivity. All four sensitive studies had at least one outcome showing efficacy of intra-articular morphine against placebo. Six studies compared intra-articular morphine with intravenous or intramuscular morphine or with intra-articular saline without a bupivacaine control. Four of the six studies showed greater efficacy for intra-articular morphine. There was no dose-response evident. No quantitative analysis of pooled data was done. We conclude that intra-articular morphine may have some effect in reducing postoperative pain intensity and consumption of analgesics. These studies had significant problems in design, data collection, statistical analysis and reporting. Trials of better methodological quality are needed for a conclusive answer that intra-articular morphine is analgesic, and that any analgesia produced is clinically useful. PMID- 9211474 TI - Blockade of nocebo hyperalgesia by the cholecystokinin antagonist proglumide. AB - In patients who reported mild postoperative pain, we evoked a nocebo response, a phenomenon equal but opposite to placebo. Patients who gave informed consent to increase their pain for 30 min received a substance known to be non-hyperalgesic (saline solution) and were told that it produced a pain increase. A nocebo effect was observed when saline was administered. However, if a dose of 0.5 or 5 mg of the cholecystokinin antagonist proglumide was added to the saline solution, the nocebo effect was abolished. A dose of 0.05 mg of proglumide was ineffective. The blockade of the nocebo hyperalgesic response was not reversed by 10 mg of naloxone. These results suggest that cholecystokinin mediates pain increase in the nocebo response and that proglumide blocks nocebo through mechanisms not involving opioids. Since the nocebo procedure represents an anxiogenic stimulus and previous studies showed a role for cholecystokinin in anxiety, we suggest that nocebo hyperalgesia may be due to a cholecystokinin-dependent increase of anxiety. PMID- 9211475 TI - The Chronic Pain Grade questionnaire: validation and reliability in postal research. AB - The Chronic Pain Grade questionnaire has been proposed as an interview administered, multi-dimensional measure of chronic pain severity in selected populations with chronic pain in the United States of America. It has not previously been tested in the United Kingdom, in self-completion form or in an unselected general population. We undertook a postal survey to assess its reliability, validity and acceptability in these circumstances, using a general practice population in Scotland, with a practice population of 11202 patients. A random sample of 400 patients aged over 18 was drawn, stratified for age, gender and receipt or non-receipt of regular prescriptions for pain-relieving medication. The dimensions and sub-scales of the Chronic Pain Grade were compared with the SF-36 general health questionnaire and questions relating to duration of any pain and attempts to seek treatment for this. The methodological approach proposed by Streiner and Norman (1989) was used to assess validity and reliability. A response rate of 76% was achieved. Cronbach's alpha was > 0.9 and item-total correlations were all high, indicating good internal consistency and reliability. Validity was confirmed by psychometric testing, including confirmatory factor analysis. Good correlations with comparable dimensions of the SF-36 general health questionnaire confirmed convergent validity. Construct validity was confirmed by testing scores against duration of pain and treatment sought for pain. We concluded that the Chronic Pain Grade questionnaire is a useful, reliable and valid measure of severity of chronic pain. It translates well into UK English and is acceptable in general population postal research. PMID- 9211476 TI - The relationship of nausea and dyspnea to pain in seriously ill patients. AB - OBJECTIVE: To describe the hospital symptom experience of seriously ill patients with common illnesses. To assess the independent association of nausea and dyspnea to level of pain. DESIGN: Cross-sectional study. SETTING: Five tertiary care academic centers in the US. PARTICIPANTS: 1556 patients admitted between June 1989 and June 1991 in SUPPORT (Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments) who answered questions concerning frequency and severity of pain, nausea and dyspnea, and the depression and anxiety subscales of the Profile of Mood States. METHODS: Seriously ill patients were interviewed a median of 8 days after hospitalization concerning the frequency and severity of their pain, nausea and dyspnea. Frequencies of symptoms and symptom combinations were tabulated. Ordinal logistic regression was used to test the independent association of level of pain with nausea and dyspnea. The analysis was adjusted for 22 additional demographic, psychological, chronic and acute illness measures. RESULTS: At least some level of anxious mood was reported by 85.2% of patients, depressed mood by 72.3% of patients, pain by 51.2% of patients, dyspnea by 48.6% of patients and nausea by 23.9% of patients. At least some degree of one of the five symptoms was reported by 94.2% of patients. Multivariable analysis controlling for demographic, psychological, and chronic and acute illness variables revealed that nausea and dyspnea were independently related to level of pain. Compared to patients without these symptoms, patients who reported mild (level 2), moderate (level 3), more severe (level 4) and very severe (level 5) nausea had odds ratios (OR) for higher levels of pain of 1.62 (1.24, 2.12) (95% confidence interval), 2.36 (1.39, 4.00), 2.57 (1.29, 5.12) and 2.22 (1.08, 4.53), respectively. Compared to patients without these symptoms, patients who reported mild (level 2), moderate (level 3), more severe (level 4) and very severe (level 5) dyspnea had odds ratios (OR) for higher levels of pain of 1.49 (1.17, 1.90), 1.92 (1.21, 3.04), 2.73 (1.83, 4.07) and 1.95 (1.39, 2.73), respectively. CONCLUSIONS: Seriously ill patients have a high symptom burden. Patients who have nausea and dyspnea experience more pain than patients without these symptoms, even after adjustment for depression, anxiety, disease type, disease severity and demographic and psychological measures. The causal association between these symptoms and pain remains to be determined. Though pain may cause dyspnea and nausea, the intriguing possibility remains that, in addition to relieving suffering, treating dyspnea and nausea may relieve pain. PMID- 9211477 TI - Blockade of calcium channels can prevent the onset of secondary hyperalgesia and allodynia induced by intradermal injection of capsaicin in rats. AB - Intradermal capsaicin injection in humans results in primary hyperalgesia to heat and mechanical stimuli applied near the injection site, as well as secondary mechanical hyperalgesia (increased pain from noxious stimuli) and mechanical allodynia (pain from innocuous stimuli) in an area surrounding the site of primary hyperalgesia. This study in rats tested the hypothesis that the secondary hyperalgesia and allodynia observed following intradermal injection of capsaicin was dependent upon activation of voltage sensitive calcium channels in the spinal cord. Responses to application of von Frey filaments of 10 mN and 90 mN bending forces were tested in all rats before and after injection of capsaicin into the plantar surface of a hindpaw. Animals were pretreated with L-type (nifedipine), N type (omega-conotoxin GVIA) or P-type (omega-agatoxin IVA) calcium channels blockers through a microdialysis fiber implanted in the spinal dorsal horn prior to the injection of capsaicin. None of the calcium channel blockers had any affect on normal sensory or motor responses. However, all three blockers dose dependently prevented the development of secondary mechanical hyperalgesia and allodynia. The threshold to mechanical stimulation with von Frey filaments was also increased significantly in animals treated with these calcium channel blockers when compared to articial cerebrospinal fluid control animals. These data suggest that calcium channels are important for the development of mechanical hyperalgesia and allodynia that occurs following capsaicin injection. PMID- 9211478 TI - The effects of G-protein and protein kinase inhibitors on the behavioral responses of rats to intradermal injection of capsaicin. AB - This study was designed to assess the role of G-proteins and protein kinases in the spinal cord in the behavioral manifestations induced by intradermal injection of capsaicin in rats. A microdialysis fiber was implanted in the spinal cord dorsal horn for administration of G-protein and protein kinase inhibitors to decipher the role of signal transduction cascades in mechanical allodynia induced by intradermal injection of capsaicin. Animals were tested for responses to graded mechanical stimuli using von Frey filaments and for responses to radiant heat stimuli outside the area of injection. The present study demonstrated that intradermal injection of capsaicin results in changes consistent with secondary mechanical allodynia without secondary heat hyperalgesia. Infusion of a G-protein inhibitor (GDP-beta-S), a general protein kinase inhibitor (H7), or selective inhibitors of protein kinase C (NPC15437), protein kinase A (H89), or protein kinase G (KT5823) into the spinal cord dorsal horn reversed the mechanical allodynia induced by intradermal injection of capsaicin in a dose-dependent manner by increasing the threshold to mechanical stimulation towards baseline. This suggests that multiple signal transduction pathways in the spinal cord are involved in the secondary allodynia that occurs following activation of C-fiber afferents by capsaicin. PMID- 9211479 TI - Characterisation of capsaicin-induced mechanical hyperalgesia as a marker for altered nociceptive processing in patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is characterised by pain and tenderness not only over inflamed or damaged joints, but also over apparently normal tissues. Experimental models suggest that these features results from changes of sensitivity within both peripheral and central neurones, but direct evidence from human disease is lacking. At present, most clinical studies have evaluated overall pain experience rather than activity within components of the nociceptive pathway. Therefore, the aim of this study was to assess the use of a capsaicin-based technique to quantify changes of neuronal sensitivity in patients with RA. First 20 microliters of capsaicin in solution (0.03 mg/ml) was applied topically for 30 min to apparently normal skin on the forearm of control subjects and patients with RA. The subsequent development of mechanical hyperalgesia to pinprick stimuli was then measured at various time points using a 74.4-mN von Frey hair. The relationship between the area of hyperalgesia and a number of clinical measures was determined. Capsaicin-induced mechanical hyperalgesia was found to decline with age in normal subjects (r = 0.47, P < 0.01). The development of hypearlgesia had a similar time course in normal subjects and patients with RA. The maximum area of hyperalgesia, however, was substantially larger in 35 RA patients; 254.3 +/- 20.7 cm2, compared with 35 normal controls; 109 +/- 7.5 cm2 (P < 0.001). An association was apparent between hyperalgesic area and a composite score of joint tenderness (r = 0.47, P < 0.01), but not with overall pain score or a systemic marker of inflammation. These results provide evidence for enhanced sensitisation of a population of sensory fibres in RA. Peripheral sensory activity over the forearms of rheumatoid patients has previously been shown to be normal and the results suggest the presence of enhanced central mechanisms in this disorder. The correlation between capsaicin-induced hyperalgesia and joint tenderness in the RA patients implies that joint symptoms arise partially as a result of central, and not exclusively peripheral, factors. The study supports the use of capsaicin-based techniques to explore nociceptive mechanisms in clinical disorders characterised by chronic pain. PMID- 9211480 TI - Pain threshold variations in somatic wall tissues as a function of menstrual cycle, segmental site and tissue depth in non-dysmenorrheic women, dysmenorrheic women and men. AB - Pain symptoms of many disorders are reported to vary with menstrual stage. This study investigated how pain thresholds to electrical stimulation of the skin, subcutis and muscle tissue varied with menstrual stage in normal women and compared these variations with those in women with dysmenorrhea and in healthy men at matched intervals. Thresholds of the three tissues were measured four times during the course of one menstrual cycle at four sites. Two of the sites were on the abdomen within the uterine viscerotome (abdomen-rectus abdominis, left and right) and two were outside it on the limbs (leg-quadriceps, arm deltoid). Calculated from the beginning of menstruation (day 0), the menstrual phases studied were menstrual (days 2-6), periovulatory (days 12-16), luteal (days 17-22) and premenstrual (days 25-28). Spontaneous pain associated with menstruation was measured from diary estimates on a VAS scale. Whereas the highest thresholds always occurred in the luteal phase regardless of segmental site or stimulus depth, the lowest thresholds occurred in the periovulatory stage for skin, whereas those for muscle/subcutis occurred perimenstrually. Dysmenorrhea accentuated the impact of menstrual phase. For non-dysmenorrheic women menstrual trends were significant only in abdominal muscle and subcutis, but for dysmenorrheic women the trends were also significant in abdominal skin and in limb muscle and subcutis. Dysmenorrhea also lowered thresholds mainly in muscle and sometimes in subcutis, but never in skin, with the greatest hyperalgesic effects in left abdominis muscle. Abdominal sites were more vulnerable to menstrual influences than limb sites. Muscle thresholds, but not skin or subcutis thresholds, were significantly lower in abdomen than in limbs, particularly in dysmenorrheic women. The amount of abdominal muscle hyperalgesia correlated significantly with the amount of spontaneous menstrual pain. Only minor sex differences were observed for pain thresholds of the arm and leg, but there was a unanimous refusal by men, but not by women, to be tested at abdominal sites. These results indicate that menstrual phase, dysmenorrhea status, segmental site, tissue depth and sex all have unique interacting effects on pain thresholds, thus adding more items to the lengthy and still-growing list of biological factors that enter into an individual's judgment of whether or not a stimulus is painful. PMID- 9211481 TI - The effect of old age on the disposition and antinociceptive response of morphine and morphine-6 beta-glucuronide in the rat. AB - The aims of this study were to examine the effect of old age on the pharmacokinetics of morphine and morphine-6 beta-glucuronide (M6G) and their relationships to antinociceptive activity. Morphine (21.0 mumol/kg) or M6G (21.7 mumol/kg) were administered s.c. to young adult and aged male Hooded-Wistar rats. Antinociceptive effect was measured by the tail-flick method at various times up to 2.5 h or 6.5 h after morphine or M6G administration, respectively, and concentrations of morphine, morphine-3 beta-glucuronide (M3G) and M6G in plasma and brain were determined by HPLC. Creatinine clearance was significantly lower by 33% or 21% in aged compared to young adult rats receiving morphine or M6G, respectively. After morphine administration, the areas under the (i) antinociceptive effect-time curve, (ii) plasma morphine concentration-time curve, and (iii) brain morphine concentration-time curve were not different between young adult and aged rats. However, the AUC for plasma M3G was five-fold higher in the aged relative to young adult rats, which could not be accounted for by only a 33% lower creatinine clearance. M6G was not detected in any plasma or brain sample from rats administered morphine and no M3G was detected in brain. For M6G administration, the areas under the (i) antinociceptive effect-time curve, and (ii) plasma M6G concentration-time curve were 1.8- and 1.6-fold higher in aged compared to young adult rats, respectively. Concentrations of M6G in brain were below the limit of quantification. No morphine or M3G was detected in any of the plasma or brain samples of rats administered M6G. The results demonstrate no change in morphine antinociception and pharmacokinetics with age, and suggest that blood-brain barrier permeability and reception sensitivity to morphine are not altered in aged rats. Accumulation of M3G in plasma of aged rats is probably due to diminished renal clearance of M3G in addition to a reduction in the biliary excretion of M3G. The heightened sensitivity of the aged rats to M6G is probably due to the observed altered kinetics of M6G rather than a pharmacodynamic change. PMID- 9211482 TI - Response to Moffett, Richardson, Frost and Osborn, PAIN, 67 (1996) 121-127. PMID- 9211483 TI - Reply to Veldman and Goris, PAIN, 64 (1996) 463-466. PMID- 9211484 TI - Response to: Back pain in the work place, PAIN, 65 (1996) 111-115. PMID- 9211485 TI - Comments on Craig et al., PAIN, 67 (1996) 285-289. PMID- 9211486 TI - Comment on Bruehl et al., PAIN, 67 (1996) 107-114. PMID- 9211487 TI - Diagnostic criteria for chronic pancreatitis by the Japan Pancreas Society. PMID- 9211488 TI - A two-step enriched-nested PCR technique enhances sensitivity for detection of codon 12 K-ras mutations in pancreatic adenocarcinoma. AB - Mutations at codon 12 of the K-ras gene have been detected in pancreatic adenocarcinomas by a variety of techniques. A few of these techniques are very sensitive, identifying the mutations in 96-100% of cases. However, these sensitive techniques are labor intensive, utilizing multistep processing and radioactive material. Much simpler techniques, involving nonradioactive single step polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) have been employed to detect K-ras mutations at codon 12 in pancreatic adenocarcinomas. However, the low sensitivity of these single-step PCR/ RFLP techniques is unacceptable. A simple and nonradio-active PCR/RFLP-based method for detection of K-ras codon 12 mutations in formalin-fixed, paraffin-embedded tissue sections of pancreatic adenocarcinoma is described and compared to the traditional PCR technique. K-ras gene mutations at codon 12 were detected by a modified two-step enrich-nested PCR (EN-PCR)/RFLP method, and their existence was confirmed by direct DNA sequencing analysis of the product. When the two-step EN PCR/RFLP technique was compared to the single-step PCR/RFLP method, K-ras codon 12 mutations were detected in 100% of pancreatic adenocarcinomas (15/15) with the EN-PCR/RFLP method, while half as many (9/15) were detected with the single-step PCR/RFLP method. This study demonstrates that the sensitivity of the simple two step EN-PCR/RFLP technique is comparable to that of the more complex methods for detecting K-ras mutations at codon 12 in formalin-fixed, paraffin-embedded tissue sections of pancreatic adenocarcinoma and its sensitivity is superior to that of the single-step technique. PMID- 9211490 TI - c-met expression in pancreatic cancer and effects of hepatocyte growth factor on pancreatic cancer cell growth. AB - Hepatocyte growth factor (HGF) is a widely expressed growth factor secreted by cells of mesenchymal origin, which has been shown to be involved in growth processes of multiple cell types. The HGF receptor, the product of the c-met protooncogene, is expressed mainly by epithelial cells. Increased expression of the HGF receptor has been observed in various tumors. To investigate the expression of the HGF receptor in the pancreas, we analyzed rat and human normal tissue and pancreatic carcinoma by Western blot analysis. We observed weak expression of c-met reactivity in normal pancreas but markedly enhanced expression in both rat and human pancreatic cancer. To test the possibility that HGF could act as a growth factor on pancreatic carcinoma, the effects of HGF on DNA synthesis in a rat and two human pancreatic carcinoma cell lines were analyzed. HGF induced dose-dependent [3H]thymidine incorporation, reaching 320, 210, and 180% above unstimulated controls in AR4-2J, PancTu-1, and 818/4 cells, respectively. The activation of signal transduction pathways by HGF was further analyzed in AR4-2J cells. After stimulation, a rapid and intense increase in receptor tyrosine phosphorylation was detected. Furthermore, HGF induced a time- and dose-dependent induction of c-fos expression. The addition of tyrphostin, a specific tyrosine kinase inhibitor, prevented c-fos induction and inhibited HGF induced [3H]thymidine incorporation. In summary, our results demonstrate strongly increased HGF receptor expression in pancreatic carcinoma and support the assumption that HGF could act as a growth promoting factor on this cancer via stimulation of tyrosine kinases. PMID- 9211489 TI - Adenoviral-mediated herpes simplex virus thymidine kinase gene transfer: regression of hepatic metastasis of pancreatic tumors. AB - Pancreatic cancer is the fifth leading cause of cancer death in the United States. Most patients have obvious metastases or locally advanced disease at the time of presentation. Surgical resection does not significantly change the clinical outcome. Combination chemotherapy induces a partial response but overall survival remains low. The aim of this study was to evaluate the feasibility of adenovirus-mediated suicide gene transduction as a therapeutic approach for pancreatic cancer. A cell line was established from a murine pancreatic ductal adenocarcinoma and intrahepatic tumors were generated by inoculation of pancreatic cancer cells into the left lateral liver lobe. Transduction efficiency was characterized in vitro and in vivo. Intrahepatic tumors were treated by intratumoral adenovirus injection in combination with intraperitoneal administration of ganciclovir. Adenovirus-mediated herpes simplex virus (HSV) thymidine kinase (tk) gene expression followed by ganciclovir treatment was highly efficient in inhibiting pancreatic cancer cell proliferation in vitro. The proliferation of nontransduced cells was significantly reduced in the presence of HSV-tk expressing cells. Intrahepatic inoculation of pancreatic cancer cells leads to successful formation of solid adenocarcinomas in syngeneic recipients. Ad.RSV-tk injection of the tumor followed by intraperitoneal ganciclovir application caused highly significant tumor volume reduction and necrosis. These results indicate that transduction of the HSV-tk gene followed by ganciclovir is highly efficient for growth inhibition of hepatic metastases of pancreatic carcinoma. PMID- 9211491 TI - Different expression of transforming growth factor beta 1 in pancreatic ductal adenocarcinoma and cystic neoplasms. AB - Pancreatic neoplasms harbor different prognoses according to their histological type: a benign course for serous cystadenoma, a low malignant potential for intraductal papillary mucinous neoplasms (IPMN), and high aggressiveness for ductal adenocarcinoma (ADC). Transforming growth factor beta 1 (TGF beta 1) may regulate tumor growth. The present study analyzes and compares the expression of its precursor beta 1-latency-associated peptide (beta 1-LAP), its latent binding protein (LTBP), and its mRNA in ductal adenocarcinoma (n = 10), in IPMN (n = 8), in serous cystadenoma (n = 2), and in normal tissues (n = 5). LTBP is thought to play a strategic role in the processing and active secretion of latent TGF beta 1 and its stockage in the extracellular matrix. Localization of beta 1-LAP and LTBP was assessed by immunohistochemistry using specific antibodies and expression of TGF beta 1 mRNA by reverse-transcriptase polymerase chain reaction analysis. beta 1-LAP was only slightly expressed in normal specimens, while LTBP was not detected. beta 1-LAP was detected in the cytoplasm of neoplastic cells in 9 of 10 patients with ADC. An intense staining was present in stromal cells surrounding the neoplastic glands in all cases except in one carcinoma in situ. LTBP was detected only in stromal cells and in the surrounding extracellular matrix. In IPMN with mild-grade dysplasia and in cystadenoma, beta 1-LAP was strongly expressed in the epithelial cells, while it was poorly detected in invasive IPMN; stromal cells were poorly or not all stained by beta 1-LAP, except in invasive IPMN (n = 2). LTBP was detected in neoplastic cells of three cases with benign IPMN and two of two cases with cystadenoma, while stroma was not immunostained. TGF beta 1 mRNA was strongly expressed in most of the tumors and no difference in expression was observed between the different types of neoplasms. There is no quantitative difference in expression of TGF beta 1 in ADC and in IPMN or cystadenoma. However, the latter are able to secrete TGF beta 1 efficiently, in contrast to ductal ADC as shown by the ability of the neoplastic cells to express both beta 1-LAP and LTBP. Invasive stroma reaction was associated with enhanced beta 1-LAP and LTBP expression in stromal cells and could be mediated by TGF beta 1 via LTBP PMID- 9211492 TI - The lobular architecture of the normal human pancreas: a computer-assisted three dimensional reconstruction study. AB - Tissue specimens of the normal pancreata from three adult patients were subjected to serial sectioning and computer-assisted three-dimensional reconstruction of the lobules, blood vessels, pancreatic ducts, and islets, to establish the anatomical relation among these structures. Most lobules, although more or less imperfectly demarcated by septa, were found to receive a single duct (lobular duct), justifying their definition as "glandular lobules." However, there were also lobules receiving not only a lobular duct of their own but a small accessory duct from a neighboring lobule. On the other hand, these lobules were shown not to correspond to the territories of arterial supply, leaving no ground for defining them as "vascular lobules." PMID- 9211493 TI - Effect of aging on B-cell function and replication in rat pancreas after 90% pancreatectomy. AB - We studied the effect of aging on B-cell function and replication in depancreatized rats. Male Wistar rats, at the ages of 1, 5, and 15 months, underwent 90% pancreatectomy (Px) or a sham operation, and islet function and regeneration were examined 3 weeks later. Plasma glucose levels in 1-month-old rats reached a peak 2 weeks after Px and then declined, while those in 5- and 15 month-old rats reached significant levels as early as the day after Px or 2 days after surgery and continued to increase over the following 3 weeks. Consequently, in contrast to young Px rats, weight loss due to severe diabetes was observed in 5- and 15-month-old Px rats. Plasma glucose responses to intravenous glucose loading (0.5 g/kg body weight) were much greater in older Px rats than in younger rats. There was a marginal insulin response to glucose in 1-month-old Px rats, whereas no insulin response to glucose was observed in older Px animals. The insulin content of the residual pancreas was increased threefold in 1-month-old Px rats, but there was no increase in 5- and 15-month-old Px rats. These data demonstrate that the effect of reducing islet mass is much greater in aged rats than in young rats and that the replicatory capacity of B cells tends to diminish after adulthood has been reached. PMID- 9211495 TI - Vitamin A stimulation of insulin secretion: effects on transglutaminase mRNA and activity using rat islets and insulin-secreting cells. AB - Retinol or retinoic acid is required for insulin release. Retinoids increase transglutaminase activity, and transglutaminase has been implicated in islet insulin release. To examine whether transglutaminase could mediate effects of retinoids on insulin secretion, we measured (i) transglutaminase activity in islets from rats deficient in vitamin A or repleted with retinol or retinoic acid, (ii) transglutaminase activity in RINm5F and INS-1 insulin-secreting cells cultured in retinol or retinoic acid, (iii) mRNA for transglutaminase in RINm5F and INS-1 cells, and (iv) insulin secretion from INS-1 cells in response to retinoic acid. Islets from rats repleted with retinol or retinoic acid showed more than twice the transglutaminase activity of islets from vitamin A deficient rats. Retinoic acid increased RINm5F cells and INS-1 cell transglutaminase activity. Retinol did not increase transglutaminase activity. Transglutaminase mRNA was detected in INS-1 cells but not in RINm5F cells. Retinoic acid increased insulin secretion from INS-1 cells as observed previously in RINm5F cells. In conclusion, retinoic acid increases transglutaminase activity in both rat islets and two insulin-secreting from INS-1 cells. Transglutaminase is a candidate for mediating retinoid-induced changes in insulin secretion. PMID- 9211494 TI - Islet hormone secretion in pancreatic cancer patients with diabetes. AB - The diabetes or impaired glucose tolerance that occurs in most patients with pancreatic cancer is characterized by profound insulin resistance. Recent evidence suggests that the diabetes may result from the presence of the tumor rather than being a predisposing factor to development of the malignancy. Some islet hormones have been shown to exhibit diabetogenic effects. To investigate the potential role of these hormones in the diabetic state associated with pancreatic cancer, we measured islet hormones during fasting in pancreatic cancer patients (n = 30), patients with other malignancies (n = 43), and healthy controls (n = 25). Preoperative pancreatic cancer patients were classified as normal glucose tolerance (NGTT), impaired glucose tolerance (IGTT), non-insulin requiring diabetes (NIRD), and insulin-requiring diabetes (IRD). Nine pancreatic cancer patients were studied after tumor removal by subtotal pancreatectomy. Some preoperative pancreatic cancer patients (n = 19), postoperative patients (n = 9), and controls (n = 8) were also studied during hyperglycemia and following glucagon injection. Fasting plasma C-peptide was elevated in NIRD pancreatic cancer patients compared to controls. Fasting levels of islet amyloid polypeptide (IAPP), glucagon, and somatostatin were elevated in NIRD and IRD patients. IAPP and glucagon, but not somatostatin, normalized following subtotal pancreatectomy. During hyperglycemia, increases in C-peptide and IAPP were seen only in controls and in NGTT and postoperative pancreatic cancer patients. After glucagon infusion, IAPP levels increased in controls and nondiabetic cancer patients; C peptide levels increased in controls, nondiabetic patients, and NIRD. Responses of C-peptide and IAPP to glucagon normalized after pancreatectomy. During hyperglycemia, glucagon levels fell in all groups except IGTT patients and a decrease in somatostatin concentrations was seen in controls. PMID- 9211496 TI - Manganese superoxide dismutase: a marker of ischemia-reperfusion injury in acute pancreatitis? AB - Recent evidence has suggested that ischemia-reperfusion injury is fundamental to the pathogenesis of acute pancreatitis. This study was designed to determine whether acute pancreatitis is associated with elevated serum manganese superoxide dismutase (MnSOD), a key antioxidant enzyme, considered a marker of ischemia reperfusion injury in myocardial infarction. Thirty-four patients with acute pancreatitis had measurements of MnSOD on days 0, 2, and 5 after recruitment. The patients were recruited within 12 h of admission to hospital and had measurements of MnSOD on days 0, 2, and 5. Patients with severe acute pancreatitis had significantly elevated serum MnSOD concentrations on days 2 and 5 compared with patients with mild acute pancreatitis, but not on the day of recruitment. Elevated serum MnSOD correlated with peripheral plasma markers of lipid peroxidation (malondialdehyde) and neutrophil activation (myeloperoxidase) and was associated with decreased plasma ascorbic acid concentrations. The serial measurement of serum MnSOD may prove useful as a marker of the effectiveness of treatment designed to limit ischemia-reperfusion injury in patients with severe acute pancreatitis. PMID- 9211497 TI - Exocrine pancreatic function in rats after acute pancreatitis. AB - The present studies were performed to evaluate pancreatic exocrine function in rats during the early stage of acute pancreatitis in two models: one is edematous pancreatitis induced by four subcutaneous injections of 20 micrograms/kg body weight of cerulein at hourly intervals and the other is hemorrhagic pancreatitis induced by retrograde infusion of 0.4 ml/kg body weight of 3% sodium taurocholate (NaTc) into the pancreatic duct. Secretory studies were performed in vivo under urethane anesthesia at various times after induction of acute pancreatitis. Basal pancreatic fluid secretion was significantly elevated after induction of acute pancreatitis in both the cerulein and the NaTc models, reaching the peak level on postpancreatitic days 1 and 3, respectively. In both models of rats, a stepwise increasing dose of cerulein was unable to cause a further increase in fluid secretion above the basal level, whereas it caused a dose-dependent increase in protein output in both models, although the responsiveness and the sensitivity were markedly reduced compared with the controls. In contrast to cerulein, secretin caused a dose-dependent increase in fluid secretion in both models of pancreatitis. In cerulein-induced postpancreatitic rats, secretin also caused a dose-dependent increase in protein output and bicarbonate concentration, but it had only a small effect at certain doses in NaTc-induced postpancreatitic rats. These results indicate that basal pancreatic fluid secretion was greatly increased but the secretory response to cerulein stimulation was reduced in acute pancreatitis early after the onset but was not reduced to secretin stimulation and that protein output and bicarbonate concentration were reduced depending on the severity of pancreatitis (NaTc-pancreatitis > cerulein-pancreatitis. PMID- 9211498 TI - The effect of beta-thia-iminoprostacyclin in taurocholate acute pancreatitis in rats: the role of antecedent acute ethanol abuse. AB - The promoting effect of acute ethanol (E) abuse and protective effect of prostaglandin derivatives in acute pancreatitis (AP) remain obscure. The aim of this study was to assess the effect of previous intake of high-dose E on trypsinogen (Tn) activation and labilization of pancreatic lysosomal membranes (PLM), in taurocholate AP in rats, considering treatment with stable beta-thia iminoprostacyclin (T). In 60 male Wistar rats taurocholate AP was induced or a sham operation was performed. Half of them received 40% E (5 g/kg body weight), 6 h earlier. T (0.3 mg/kg body weight i.g.) was applied before E or before the induction of AP. Free active (FAT) and total potential (TPT) trypsin, free (F) and total (T) cathepsin B, phospholipase A2 (PLA2), and lipase (L) activities were assayed. Percentage FAT/TPT was an index of Tn activation and fractional free (% F/T) activity of cathepsin B was an index of PLM fragility. FAT increased after 12 h of AP, and in E rats this increase was even more evident. Pretreatment and treatment with T partly prevented this increase, however, this effect was abolished or limited in rats previously given E-the changes were not effected by T. PLA2 and L activities in AP were not diminished after T. The promoting effect of acute E abuse prior to AP could be dependent on augmented activation of Tn and labilization of PLM. The protective effect of T seems to be dependent on the decrease in Tn activation in pancreatitic tissue. The potential therapeutic effect of this drug in AP could be limited by previous acute E intake, as evidenced by differences in histopathological changes. PMID- 9211500 TI - Antineutrophil cytoplasmic antibodies and acute pancreatitis. PMID- 9211499 TI - Pancreatic schwannoma: report of two cases and review of the literature. AB - Solitary intrapancreatic schwannoma is a rare tumor. We present two patients with this tumor and review 13 previously reported cases from the English-language literature. While the final diagnosis was made based on pathological examination of the tumors, both computed tomography scan and magnetic resonance imaging helped establish the benign nature of the lesion, narrow the differential diagnosis, and define the anatomical locations of the small tumors. Both tumors were treated by enucleation from the surrounding pancreatic parenchyma, and both patients, after 2 years of follow-up, are alive and well. It is concluded that multimodality radiologic investigations are useful in the workup of unusual pancreatic masses. In addition, based on the known biologic behavior of schwannomas occurring elsewhere in the body, simple enucleation, rather than more radical resection, is likely to be adequate therapy for these tumors. PMID- 9211501 TI - Sac-like pouches in Blastocystis from the house lizard Cosymbotus platyurus. PMID- 9211502 TI - Theileria-mediated constitutive expression of the casein kinase II-alpha subunit in bovine lymphoblastoid cells. AB - Theileria-infected cells are induced to undergo a transformation that is reversible, since their proliferation is inhibited after elimination of the schizonts by the theilericidal drug buparvaquone. The molecular mechanisms of the transformation remain unknown. The experiments described in the present report deal with the role of casein kinase (CK) II, a serine/threonine protein kinase, in the permanent proliferation of the parasitized cells and show that the CK II alpha subunit is expressed in both T. annulata- and T. parva-infected cells and that its expression is closely related to the presence of the parasites in the host-cell cytoplasm. Thus, elimination of the schizonts by buparvaquone leads to the inhibition of CK II-alpha subunit mRNA expression without affecting the expression of actin. Cells treated with 5,6-dichloro-1-beta-D ribofuranosylbenzimidazole (DRB) are inhibited in a dose-dependent manner from under-going DNA synthesis as measured by [3H]-thymidine incorporation and from expressing CK II. Furthermore, a host-cell-specific CK II-alpha antisense inhibits DNA synthesis in a dose-dependent manner. In the present study, 6 microM antisense reduced [3H]-thymidine incorporation by Theileria-infected bovine cells to about 50%. Using a primer derived from T. parva CK II, we detected a parasite specific CK II mRNA in T. parva-infected cell lines. Interestingly. DRB also inhibited the expression of the parasite-specific CK II. However, to date we have not detected a target sequence for this primer in T. annulata schizonts. PMID- 9211503 TI - Detection of sperm within Gyrodactylus (Platyhelminthes, Monogenea) tissues using fluorescence and transmission electron microscopy. AB - A rapid fluorescent staining method demonstrating spermatozoa within gyrodactylid monogeneans is described. Gyrodactylids fixed and stained in 2% acetic acid containing 1 microgram ml-1 bisBenzimide (Hoechst 33258) were viewed using epifluorescence microscopy. In addition to staining sperm in the testis, seminal vesicle and seminal receptacle, the technique highlighted sperm in the gut and cytoplasmic lining of the uterus, locations from which they had not previously been recorded. The technique was more rapid than transmission electron microscopy (TEM), which was used to confirm the observations. Fluorescence microscopy provided an overview of sperm orientation and distribution that could otherwise be obtained only from serial ultrathin sections. Using the fluorescence technique and/or TEM we have located migrating sperm in the tissues of Gyrodactylus turnbulli, G. bullatraudis and G. gasterostei, although these sperm do not appear to enter the tissues of embryos in utero. The technique can be used to study insemination patterns in gyrodactylid populations and in experimental studies of gyrodactylid reproduction. PMID- 9211504 TI - Ceratomyxa sparusaurati (Protozoa:Myxosporea) infections in cultured gilthead sea bream Sparus aurata (Pisces:Teleostei) from Spain: aspects of the host-parasite relationship. AB - Ceratomyxa sparusaurati infection was studied in gilthead sea bream from different mariculture systems of Spain. Culture conditions were found to influence the infection dynamics. Total prevalences ranged from 0 in a closed system to 59.9% in a Mediterranean semi-intensive farm. Prevalence was significantly lower in summer in a restricted group from the latter system, but no clear seasonal pattern was observed in the remaining fish groups in any system. A statistically significant dependence between infection prevalence and host weight was observed in some fish groups. Different degrees of histopathological damage were noted in the infected gallbladders, mainly involving swelling, vacuolization, and sloughing of the epithelial cells. Transmission electron microscope observations of the infected tissues demonstrated a characteristic protrusion of the epithelial cell surface and mitochondrial alterations. In fish from stocks showing external disease signs and trickling mortalities, parasite stages invaded other organs and were responsible for cell-mediated host reaction and important tissue injuries. Therefore, C. sparusaurati can be considered a potential threat for some Sparus aurata-growing systems. PMID- 9211505 TI - The anterior central nervous system of male Onchocerca volvulus (Nematoda:Filarioidea) as a target for chemotherapy: results of an electron microscopy study. AB - An electron microscopy study was performed to evaluate the feasibility of the use of the anterior nerve ring of male Onchocerca volvulus for the assessment of early drug effects. Worms were exposed to new and known compounds at reasonable concentrations of 1 microM and less for 6, 12, 18, and 36 h in an established in vitro system. The anterior end of the filariae up to a length of 1 mm was examined and the morphological findings were compared with motility and reduction of a tetrazolium sat to formazan by live but not dead worms. The nerve fibers were more susceptible to the chemotherapeutic intervention then the other tissues in the anteriormost part of the worms. The alterations depended on the duration of exposure and the chemical nature of the compounds used. Morphological changes in the nervous tissue and the inhibition of motility and formazan production corresponded well for the arsenical mel w, used as an active standard, two pyrimidinyl-guanidines (PD 105482 and PD 105666), and an imidazolinylhydrazone (WR 251993). PMID- 9211506 TI - Supporting experimental evidence of host specificity among southern African polystomes (Polystomatidae:Monogenea). AB - Although monogeneans of anurans are generally regarded as host-specific, there is a lack of conclusive experimental evidence. Infection and cross-infection experiments were conducted with oncomiracidia of Polystoma australis and P. marmorati. In a series of experiments, oncomiracidia were given a choice between natural and substitute host tadpoles. Oncomiracidia of P. australis became established in substitute hosts but showed a preference for the natural host, whereas the oncomiracidia of P. marmorati showed a strong and statistically significant preference for the natural host. The results indicated that although the oncomiracidia of southern African polystomes showed a strong preference for their natural hosts, not all parasites exercised the same degree of host specificity. PMID- 9211507 TI - Prepatent periods of different Oesophagostomum spp. isolates in experimentally infected pigs. AB - To define prepatent periods of different Oesophagostomum spp. isolates we carried out two separate experiments, one using two monospecific laboratory isolates and another using laboratory isolates as well as isolates obtained from pig herds having different management systems and with different anthelmintic treatment histories. Pigs were inoculated with 1,000-2,000 infective larvae. Fecal samples were collected daily beginning on days 15 and 16 postinoculation (p.i.). Fecal cultures were set up at different times to yield larvae that could be identified by DNA analyses. All pigs started to excrete eggs on days 18-24 p.i. The mean prepatent period was 20.2 +/- 1.4 days, with no significant difference being observed between species and isolates. Prepatent periods of 17-19 days were found for the monospecific laboratory isolates of O. dentatum and O. quadrispinulatum. These findings conflict with parasitology textbooks; therefore, suggestions as to the possible reasons for the observed short prepatent periods are given. PMID- 9211508 TI - Permissiveness of two African wild rodents, Mastomys huberti and Arvicanthis niloticus, to Schistosoma intercalatum: epidemiological consequences. AB - The compatibility between Schistosoma intercalatum (Cameroon) and two wild rodents commonly found in Africa. Mastomys huberti (the multimammate mouse) and Arvicanthis niloticus (the Nile rat) was studied to determine their biological capacities to act as hosts for S. intercalatum. In both rodent species the general mean worm recovery was high (33 +/- 0.1% in M. huberti and 33.8 +/- 0.1% in A. niloticus) and worm mortality was very low from 6 to 20 weeks postinfection; parasite maturity was reached. The high number of released eggs as well as the viability and the infectivity of the miracidia to the snail vector showed that M. huberti and A. niloticus are very permissive to S. intercalatum and may act as hosts for the human disease. PMID- 9211510 TI - Influence of PF1022A on the motility of Angiostrongylus cantonensis in vitro. AB - Our previous in vivo studies on angiostrongyliasis showed that PF1022A had stronger killing effects against female adults than against males. No killing effects were observed against young adult worms in the central nervous system. To characterize the former in vivo action of PF1022A, in vitro effects of PF1022A on the motility of Angiostrongylus cantonensis were studied directly. Few differences in the efficacy were observed between male and female worms, but dose and time-dependent inhibition was observed in adults treated with PF1022A at 10( 7)-10(-11) g/ml. PF1022A was slightly less effect we against young-adult worms than against adult worms. Minor effects of PF1022A were observed on the third stage larvae. These results suggest that selectivity against adult females in vivo could be attributable to non-neuropharmacological mechanisms and that PF1022A does not pass through the blood-brain barrier in host animals. PMID- 9211509 TI - Isolation of coccidian enteroepithelial stages of Toxoplasma gondii from the intestinal mucosa of cats by Percoll density-gradient centrifugation. AB - A method for isolation of enteroepithelial stages of Toxoplasma gondii from the intestinal mucosa of experimentally infected cats was developed using Percoll density-gradient centrifugation. Gamonts and merozoites were obtained essentially free of host-cell debris. A recovery rate of nearly 30% of the parasites in the original preparations was obtained by this method. Merozoites were separated from gamonts by filtration through a 3-micron polycarbonate filter. PMID- 9211511 TI - Eosinophilia and intracranial worm recovery in interleukin-5 transgenic and interleukin-5 receptor alpha chain-knockout mice infected with Angiostrongylus cantonensis. AB - We infected interleukin-5 (IL-5)-transgenic (IL-5-Tg) and IL-5 receptor alpha knockout (IL-5R alpha -/-) mice with Angiostrongylus cantonensis to determine the possible roles of IL-5 and eosinophils in A. cantonensis infection in mice. IL-5 Tg mice demonstrated significantly higher eosinophilia in bone marrow, blood and cerebrospinal fluid (CSF), lower intracranial worm recovery and smaller female worms than naive C3H/HeN mice. Both IL-5-Tg and C3H/HeN mice evoked antigen specific serum and CSF IgA antibody responses as early as days 5 and 7 postinfection, respectively. Prominent eosinophil infiltration was noted around intracranial worms in the subarachnoid spaces of the mouse brains; eosinophils adhering to the worm surface were degranulated. In contrast, IL-5R alpha -/- mice yielded a higher worm recovery than wild-type or heterozygous mice at day 20 postinfection and failed to provoke CSF eosinophilia. These findings indicate that A. cantonensis infection in the mouse causes IL-5 production and subsequent CSF eosinophilia, the latter probably being involved in the killing of intracranial worms. PMID- 9211512 TI - Electron microscope observations on Onchocerca ochengi and O. fasciata (Nematoda:Filarioidea). AB - The fine structure of the females and males of Onchocerca ochengi (parasitizing zebu and cattle) and of the females of O. fasciata from camels were described and compared to other filariae of the genus Onchocerca. It was shown that O. ochengi resembles O. volvulus of humans in its degree of development, while being more primitive than O. gibsoni. Besides other similarities O. ochengi attracts inflammatory cells in the way of O. volvulus and these could be a model for chemotherapeutic trials. PMID- 9211513 TI - Biochemical and immunological characterization of soluble antigens of Giardia lamblia. AB - The crude soluble antigens (CSA) of Giardia lamblia trophozoites and their analytically purified fractions were characterized biochemically and immunologically to determine the most immunogenic fraction and its localization on the parasite. Both Sephacryl S-300 column chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed the highly complex and heterogeneous nature of CSA. Gel filtration of CSA showed four fractions (FI FIV) with molecular masses of 250, 150, 110, and 10 kDa for fractions I-IV, respectively. Protein profiles of CSA demonstrated 28 Coomassie-blue bands in the range of 200-14 kDa. Similar banding patterns with fewer polypeptides were observed in the FI fraction. However, fractions II and III showed polypeptide bands in the region of 97-14 kDa. The glycoprotein nature of CSA and its fractions were demonstrated in physicochemical analysis. In antigenic activity analysis the high-molecular-weight FI antigen was found to be 8 times more immunogenic than CSA as well as the other fractions. Major differences in the immunoreactivity of CSA and FI antigens were noted at 220, 30, and 22 kDa for the FI antigen and at 205, 84, 55, 43, and 20 kDa for CSA. Some of these FI polypeptides were found to be surface-associated as revealed by immunofluorescence and immunoblot assay. These results suggest the future use of the FI antigen in the serodiagnosis of and immunoprophylaxis against giardiasis. PMID- 9211514 TI - Diffusion of microinjected markers across the parasitophorous vacuole membrane in cells infected with Eimeria nieschulzi (Coccidia, Apicomplexa). AB - Cells infected with the intracellular parasite Eimeria nieschulzi were microinjected with lucifer yellow (457 Da), biocytin lucifer yellow (850 Da) and dextranrhodamine (10,000 Da). Immediately after injection of a mixture of the markers into the host cell cytoplasm, a differential diffusion pattern was observed in trophozoites and schizonts. Lucifer yellow and biotin lucifer yellow were seen to enter the parasitophorous vacuole, whereas the dextran was excluded. Since these markers cannot permeate cell membranes, this suggests that the Eimerian parasitophorous vacuole acts as a molecular sieve. This method allows the visualization and further characterization of the parasitophorous vacuole in living cells. PMID- 9211515 TI - Ascaridia galli populations in chickens following single infections with different dose levels. AB - In all, 3 groups of 20 Lohman Brown chickens aged 1 day were orally infected with doses of 100, 500, or 2,500 embryonated Ascaridia galli eggs, respectively. After 8 weeks, egg counts (eggs per gram of feces, EPG) were determined for all animals prior to slaughter. The gastrointestinal tracts were examined for the presence of adult and immature stages of A. galli. All groups had roughly similar worm burdens and, hence, significantly different establishment rates of 14.2%, 2.9%, and 0.5%, respectively. A significantly lower mean female worm burden was seen in the high-dose group (P = 0.02), which also showed a significantly lower level of egg excretion (P = 0.01). However, fecundity (EPG per female) did not significantly differ between the groups (P = 0.55). The mean lengths of adult worms as well as the weight of the mean worm burdens were significantly smaller in the high-dose group. This study demonstrated that single infections with varying doses of A. galli eggs influenced the establishment rate, sex ratio, egg excretion, and worm size and weight but not the worm fecundity. PMID- 9211516 TI - Lead and cadmium content of two cestodes, Monobothrium wageneri and Bothriocephalus scorpii, and their fish hosts. AB - The adult cestodes Monobothrium wageneri and Bothriocephalus scorpii from the intestines of their respective final hosts, tench (Tinca tinca) caught in the river Ruhr, Germany, and turbot (Scophthalmus maximus) collected from two sampling sites on the coast of Gdansk, Poland, were analyzed for lead and cadmium by atomic absorption spectrometry. Both cestode species contained significantly higher cadmium contents than did the muscle, liver, and intestine of their fish hosts. Whereas M. wageneri also contained several times more lead than did the organs of tench, B. scorpii showed nearly the same lead burden as did the liver and intestine of turbot. Posterior sections of B. scorpii comprising gravid proglottids contained significantly higher concentrations of lead and cadmium than did the anterior proglottids. PMID- 9211518 TI - Thelazia lacrymalis (Nematoda, Spirurida, Thelaziidae): report in a horse in Germany and contribution to the morphology of adult worms. PMID- 9211517 TI - The in vitro effects of extracellular adenosine triphosphate on the ultrastructure of Trypanosoma cruzi epimastigotes. AB - Incubation for 24 h in culture medium containing 50 mM adenosine triphosphate (ATP) produces distinct alterations in the ultrastructure of Trypanosoma cruzi epimastigotes, most obvious of which is the formation of large membrane-bound vacuoles in the cytosol. These vacuoles become positive following exposure to the macromolecule horseradish peroxidase (HRP). After a 20-min chase in phosphate buffered saline (PBS) the HRP-positive vacuoles begin to separate into discrete structures such that after a 60-min chase, obvious reservosomes are identifiable. It is hypothesized that extracellular ATP causes increased permeability of the epimastigote's plasma membrane, resulting in ionic fluxes that, in turn, interfere with the normal formation of reservosomes. PMID- 9211519 TI - Cathepsin B-like activity predominates over cathepsin L-like activity in adult Schistosoma mansoni and S. japonicum. AB - Both cathepsin B-like and cathepsin L-like endopeptidase activities have been described in schistosomes, but their relative contribution to proteinolysis remains controversial. In an attempt to clarify which type of activity predominates, the selective mammalian cathepsin B inhibitor CA-074 was tested under standardized assay conditions with different preparations from Schistosoma mansoni and S. japonicum. CA-074 (0.94 microM) inhibited at least 92% and 80% of proteinolytic activity, respectively, for these species: completely inhibited bovine-spleen cathepsin B activity; but showed only marginal inhibition (4%) of rat-liver cathepsin L activity. We discuss the results with respect to previous studies and conclude that schistosome cathepsin B-like, not L-like, activity predominates. PMID- 9211520 TI - Modulation of S-100 genes response to growth conditions in human epithelial tumor cells. AB - Many new members of the S-100 genes are known to be associated with cell differentiation, malignant transformation, and cell cycle. Of the S-100 genes examined in the present study, calcyclin, calpactin I light chain and calvasculin were expressed in most human epithelial tumor cells, and their expression levels differed according to various growth conditions. Their transcribed levels differed depending on each cell line, but their expression patterns were similarly changed under growth-modulatory conditions. Their messenger RNA levels increased parallel to the S phase population of cells, and decreased at G1/G2 phases. In contrast, this expression diminished in tumor cells under growth inhibitory conditions, such as treatment with topoisomerase II inhibitor VP-16 or phorbol 12-myristate 13-acetate. PMID- 9211521 TI - Hemosiderin deposition in portal endothelial cells is a histologic marker predicting poor response to interferon-alpha therapy in chronic hepatitis C. AB - Interferon (IFN)-alpha is regarded as an efficient therapy for chronic hepatitis C, despite the fact that less than 50% of patients receiving IFN-alpha are known to show an initial biochemical response, and several adverse reactions related to this therapy are becoming a serious clinical problem. For a more efficient and safer treatment of IFN-alpha, several pretreatment factors to predict a favorable or unfavorable response to IFN-alpha therapy are now being evaluated, such as hepatitis C virus (HCV)-RNA levels in serum and the genotypes of HCV. Recently, the hepatic iron concentration has been reported to influence the outcome of IFN alpha therapy for chronic viral hepatitis. In the present study, whether hemosiderin deposition in liver is a histologic predictor of response to IFN alpha therapy was evaluated, as well as which anatomical location showing the hemosiderin deposition was more closely related to the response to this therapy. Two factors, high titer of HCV-RNA in serum and hemosiderin deposition in portal endothelial cells, were found to be predictable factors of poor response to IFN alpha therapy, and these two factors were found to be related to each other. Results showed that the hemosiderin deposition in portal endothelial vessels is an easily evaluable histologic finding, and clinicians and histopathologists are encouraged to use this finding when selecting patients with chronic hepatitis C suitable for IFN-alpha therapy. PMID- 9211522 TI - Late onset type I familial amyloidotic polyneuropathy: presentation of three autopsy cases in comparison with 19 autopsy cases of the ordinary type. AB - Clinicopathological features of three autopsy cases of extremely rare late onset type I familial amyloidotic polyneuropathy were presented and compared with 19 autopsy cases of the ordinary type. In the late onset cases, the ages at onset and at death were 27.5 and 24.5 years older, respectively, compared with the ordinary type. Also, duration of the total clinical course from onset to death was 3.7 years less than in the late onset cases. The degree of amyloid deposition was more marked in the heart of the late onset cases, causing prominent cardiac hypertrophy. It was also marked in the kidneys or thyroid of two cases, but slight to moderate in the peripheral or autonomic nervous tissues in all cases. Immunohistochemical investigation demonstrated the presence of transthyretin (TTR) as an amyloid precursor protein and of serum amyloid P-component in amyloid deposits in various organs and tissues of the late onset type. These findings, as well as serum levels of variant TTR, were similar to those of the ordinary type. These results suggest that there are some factors other than the amyloid precursor protein that effect the degree of amyloid deposition. PMID- 9211523 TI - Morphological characteristics of Epstein-Barr virus-related early gastric carcinoma: a case-control study. AB - A strong association between Epstein-Barr virus (EBV) and gastric carcinoma has been demonstrated by the uniform presence of EBV in all carcinoma cells, episomal monoclonality, elevated antibodies, and a unique 'lace pattern' in the mucosa. The present study is concerned with morphological changes of intramucosal carcinoma and submucosal invasion, reactive lymphocytes, and with atrophy and intestinal metaplasia of the surrounding mucosa. Fifty-two EBV-positive early gastric carcinomas were matched by age, sex, and site of tumor with 103 EBV negative carcinomas, all of which had previously been examined by serial cut sections of the tumors and surrounding mucosa. Epstein-Barr virus involvement was strongly associated with lace pattern morphology as demonstrated previously, and with lymphocytic infiltration in and around the tumor nests in the mucosa. The infiltrating lymphocytes in the tumor nests were mainly composed of CD8+ T lymphocytes. Lymphoepithelioma (LE)-like carcinoma, was observed in the submucosal portions of 13 of 31 EBV-positive cases with such invasions, including 12 of 29 with lace pattern morphology in the mucosa. The majority of the surrounding gastric mucosa showed moderate to severe atrophy with marked depletion of parietal cells, complete-type intestinal metaplasia in EBV-positive cases, and Helicobacter pylori (H. pylori) infection for both EBV-positive and EBV-negative cases. It is suggested that EBV infection may occur in the atrophic gastric epithelial cells associated with intestinal metaplasia and H. pylori infection, leading to the development of carcinoma. Such cancers show lace pattern and marked lymphocytic reaction in the mucosa, with some tendency for histological change and lymphocytic reaction during the invasive process without lymph node metastasis. PMID- 9211524 TI - A human gall-bladder signet ring cell carcinoma cell line. AB - To date, very few reports of the establishment of gall-bladder cancer cell lines have appeared, although many cancer cell lines of various kinds have been established. On the other hand, no reports could be found on signet ring cell carcinoma cell lines derived from the gall-bladder and only five cell lines from the stomach. A human gall-bladder cancer cell line (FU-GBC-2) was established in tissue culture from the ascitic fluid of a 69-year-old Japanese female patient. The tumor cells growing in tissue culture exhibited the morphological characteristics of signet ring cells in phase contrast and electron microscopy. The population doubling time was 43 hours. Heterotransplantation was succeeded by inoculation into the dermis of BALB/c nude mice. An immunocytochemical study showed that most of the cultured cells were positive for carcinoembryonic antigen, CA19-9 and epithelial membrane antigen, but negative for vimentin. The modal chromosome number was 120 with a range of 100-124. Flow cytometry showed an aneuploidy pattern in the cultured cells at passage 30. Markedly amplified c-myc oncogene was observed by Southern blot analysis. This cell line may be useful in the study of the morphological and biological characteristics of signet ring cell carcinoma and gall-bladder adenocarcinoma. PMID- 9211525 TI - Morphologic characteristics of N-methyl-N-nitrosourea-induced retinal degeneration in C57BL mice. AB - Morphologic characteristics of retinal degeneration induced by a single systemic administration of N-methyl-N-nitrosourea (MNU) in mice was investigated. The aim was to characterize the MNU-induced retinal lesions in mice and compare them with human retinitis pigmentosa. A dose of 60 mg/kg body weight MNU, injected intraperitoneally into male and female C57BL mice, evoked progressive retinal degeneration in all treated mice, while control mice remained normal. An early change was photoreceptor apoptosis followed by infiltration of macrophages and swelling of the pigment epithelial cells with phagosomal inclusions for apoptotic photoreceptor cell removal. Loss of the majority of photoreceptor cells occurred within a week. Then, Feulgen-positive corpuscles, indicative of an aggregation of degenerative photoreceptor elements, vitread the outer limiting membrane were surrounded by Muller cell processes, and the duplication of the pigment epithelial cells sclerad the outer limiting membrane were seen 2 and 3 weeks after the treatment. Finally, the Feulgen-positive corpuscles disappeared and Muller cell processes were in direct contact with the continuous lining of the single layer of pigment epithelial cells. As in retinitis pigmentosa in humans, the primary event was loss of photoreceptor cells by apoptosis, but the migration of the pigment epithelial cells within the retina was not seen in the present model. PMID- 9211526 TI - Epstein-Barr virus (EBV)-induced CD30+ natural killer cell-type malignancy resembling malignant histiocytosis: malignant transformation in chronic active EBV infection associating hypogammaglobulinemia. AB - A 27-year-old male suffered from Epstein-Barr virus (EBV)-related liver dysfunction with persistent hypogammaglobulinemia. IgG titers to EBV antigens were significantly high, while other hepatitis markers were negative. Liver biopsy disclosed active intralobular inflammation. Two years later, he manifested persistent fever, leukopenia, effusions and hypoproteinemia, and his general condition worsened progressively. The peripheral blood small lymphocytes predominantly expressed natural killer (NK)-like phenotypes (CD2+, CD7+, CD16+, CD56+). Hepatosplenomegaly and marked elevation of serum lactic dehydrogenase were observed. He died of respiratory failure at the age of 29. At autopsy, the liver (2190 g), spleen (860 g), small bowel and mesenteric lymph nodes showed massive infiltration of large atypical lymphoid cells in close association with hemophagocytic histiocytes. Involvement was mildly noted also in the bone marrow, lungs, gall-bladder and kidneys. The atypical cells belonged to CD30+ activated NK-type cells expressing CD2, cytoplasmic CD3 epsilon, CD7, CD45RO, CD56, HLA-DR and HLA-DQ. T cell receptors (TCR), surface CD3, CD4, CD5 and CD8 were not expressed. Epstein-Barr virus-related small nuclear RNA (EBER1) and Epstein-Barr virus-associated nuclear antigen 1 were detected in the nuclei of a significant number of atypical cells, while EBV-related latent membrane protein-1 was negative. EBER1 was also identified in the nuclei of non-neoplastic small lymphocytes at both biopsy and autopsy. Monoclonal integration of the EBV genome into the lymphoma cells was shown by Southern blot analysis. Clonal rearrangement of TCR was undetectable. Roles of chronic active EBV infection in the development of NK cell-type malignancy resembling malignant histiocytosis are discussed. PMID- 9211527 TI - Juxtaglomerular cell tumor of the kidney: report of a non-functioning variant. AB - A case of a juxtaglomerular cell tumor (JCT) in a 46-year-old man is reported. The tumor, 2.4 cm at its greatest dimension, was incidentally detected by ultrasonography. Although histological, immunohistochemical, and electron microscopic examinations revealed typical features of a JCT, the patient had no history of hypertension or hypokalemia. This is the first report of a non functioning JCT in the literature. PMID- 9211528 TI - Dedifferentiated chondrosarcoma of the rib with a malignant mesenchymomatous component: an autopsy case report. AB - A rare variant of dedifferentiated chondrosarcoma with malignant mesenchymomatous component in a 57-year-old male is reported. The patient presented with a posterior mediastinal mass arising from the left eighth and ninth ribs showing well differentiated, low-grade chondrosarcoma. Five years later, local recurrence occurred and an excised specimen also showed the same histological features as the primary tumor. Another 6 years later, the tumor recurred and metastasized to the multiple organs, the patient dying 4 months later. Autopsy revealed that the recurrent and metastatic tumors showed malignant mesenchymomatous 'dedifferentiation' of chondrosarcoma composed of rhabdomyosarcoma, angiosarcoma, chondrosarcoma, osteosarcoma, and leiomyosarcoma, in addition to fibrosarcomatous areas. Although the less differentiated component of dedifferentiated chondrosarcoma usually shows the histological features of malignant fibrous histiocytoma and fibrosarcoma, multilineage differentiation can occur in that component. The phenomenon of 'dedifferentiation' in chondrosarcoma and the relationship to and distinction from malignant mesenchymoma of soft tissue and bone are discussed. PMID- 9211529 TI - Spindle cell carcinoma of the breast: a case report and an immunohistochemical study including p53 and Ki-67 expression. AB - A rare case of spindle cell carcinoma (SpCC) of the breast occurring in a 51-year old Japanese woman is reported. A firm and well-circumscribed tumor, measuring 9 x 8.5 x 8.5 cm, was located on the upper lateral region of the right breast. Microscopically, the tumor consisted of sheets of both malignant spindle cells and poorly differentiated ductal carcinoma containing squamoid islands with gradual transition to the spindle cell component. The immunocytochemical expression of epithelial markers was recognized in the spindle cells, as well as in the carcinomatous cells. Moreover, the spindle cell component expressed vimentin, alpha-smooth muscle actin and S-100 protein. Ultrastructurally, in addition to the features of adenocarcinoma, squamous or myoepithelial differentiation was confirmed in the spindle cell component. These findings thus suggest an epithelial origin with squamous differentiation and myoepithelial participation in the genesis of SpCC. In a comparative study, the expression of p53 protein and Ki-67 as a proliferation marker in each component of this tumor was also investigated. The mean p53 labeling index (LI) in both the carcinomatous and spindle cell area was similar, however the mean MIB-1 LI in the spindle cell area was significantly higher than that in the carcinomatous area. The results indicate that p53 overexpression is involved in the tumorigenesis of both components in the SpCC, and the spindle cell component shows a higher degree of proliferative activity than the carcinomatous component. PMID- 9211530 TI - Endometrial low-grade stromal sarcoma with ovarian sex cord-like differentiation: report of two cases with an immunohistochemical and flow cytometric study. AB - Two cases of endometrial low-grade stromal sarcoma with ovarian sex cord-like differentiation occurring in a 39-year-old woman and a 42-year-old woman are presented. Both tumors, which were intramyometrial and measured 7.5 cm and 7.0 cm in greatest diameter, respectively, showed a multinodular, ill-demarcated, and yellowish white cut-surface. Histologically, most parts of the tumors were composed of trabecular, cord-like, or plexiform arrangements that were reminiscent of the growth pattern seen in ovarian sex cord tumors. Features of conventional endometrial low-grade stromal sarcoma were only focally observed. The tumors showed infiltrative margins and lymphatic invasion. The tumor cells were positive for vimentin, desmin, alpha-smooth muscle actin, and muscle actin (HHF35). The tumors were also positive for both estrogen and progesterone receptors. Both tumors were DNA diploid as determined by flow cytometry. One patient had recurrences, including osteolytic lesions in the pelvic bones, but had no evidence of recurrence or metastasis 11 months after the last surgery. The other patient had no evidence of tumor in a limited follow-up. Familiarity with the neoplasm and other uterine mesenchymal tumors with ovarian sex cord-like differentiation by gynecologists and pathologists is essential in avoiding misdiagnosis because adjuvant hormonal therapy may be effective in treating low grade stromal sarcomas. PMID- 9211531 TI - A new aspect of gastric metaplasia in Crohn's disease: bidirectional (foveolar and pyloric) differentiation in so-called 'pyloric metaplasia' in the ileum. AB - Mucus-secreting cells found at the site of ileac ulceration in Crohn's disease have been described as 'pyloric metaplasia'. Using mucin-histochemical methods and immunohistochemical stainings for Ki-67 antigen and foveolar-type mucin (M1) of the stomach, the characteristics of this type of metaplasia were studied in surgically resected ileac specimens from two Japanese patients with Crohn's disease. Not only pyloric-type cells but also foveolar-type cells were demonstrated; often displaying an organoid growth of the normal stomach mucosa. Stem cells of the ileac crypt may differentiate potentially to intestinal-, pyloric- and also to foveolar-type cells. The term 'pyloric metaplasia' is not appropriate and 'gastric metaplasia' should be used when describing this type of metaplasia. PMID- 9211532 TI - Measurement of urinary vanillylmandelic acid (VMA) and homovanillic acid (HVA) for diagnosis of neural crest tumors. PMID- 9211533 TI - Oxygen parameters in the presence of hemoglobin H. PMID- 9211534 TI - Pediatric oncology in Argentina: a historical overview. PMID- 9211535 TI - History of pediatric hematology-oncology in the Czech Republic. AB - Specialized treatment for children with hematologic and oncologic diseases in the Czech Republic has a relatively short history. Dr. Josef Koutecky in 1964 was the first to organize the discipline of oncologic treatments and started a department that has since grown to include six other senior members. They treat over 250 new patients a year (approximately 150 malignant tumors) with an incidence similar to that in other European nations. Most are treated on cooperative group protocols with other centers in Europe and North America. Over 30 children also receive bone marrow transplants yearly from this group. Hematologic diseases including leukemia were first treated by D.r. O. Hrodek since the late 1950s in the Second Department of Pediatrics in Prague. Eight other centers in the Czech Republic besides Motol treat children with leukemia. Since 1985 they have followed the Berlin-Frankfurt-Munster (BFM) protocols for patients with acute lymphoblastic leukemia (ALL) and acute myeloblastic leukemia (AML). The Motol Hematology Section does about 20 transplants a year. New molecular biology laboratories in both sections aid the diagnosis and follow-up of the patients. PMID- 9211536 TI - University of Istanbul, Institute of Oncology. PMID- 9211537 TI - Tissue oxygenation in patients with hemoglobinopathy H. AB - To evaluate the degree of tissue hypoxia in patients with hemoglobinopathy H disease, whole blood oxygen affinity was estimated and analyzed in 33 patients. Twenty patients with iron deficiency anemia, matched for degree of anemia, served as controls. The results were as follows: Whole blood oxygen equilibrium curves of patients with HbH disease are biphasic because of a combination of the rectangular hyperbolic curve of HbH and the normal sigmoid curve of HbA and are shifted toward the left (P50 3.66 +/- 0.33 kPa). Patients with iron deficiency anemia have right-shifted oxygen equilibrium curves (P50 4.02 +/- 0.13 kPa) compared with normal. Oxygen release to the tissues in HbH disease is decreased (1.4 +/- 0.3 mmol/L) as compared with iron-deficient patients (1.6 +/- 0.2 mmol/L) with a similar degree of anemia. Red cell indices vary between the two groups. In patients with HbH disease the mean corpuscular hemoglobin concentration was 268 +/- 17 g/L as compared with 294 +/- 18 g/L in iron deficiency anemia. These findings indicate that whole blood oxygen affinity is a reliable index of tissue oxygenation in patients with hemoglobinopathy H. PMID- 9211538 TI - Febrile complications in the first 100 days after bone marrow transplantation in children: a single center's experience. AB - One hundred fifty-six episodes of fever occurred in 102 children during the first 100 days after bone marrow transplantation (BMT) performed at a single institution: fever of undetermined origin (FUO), 40.3%; septicemia, 7.1%; pneumonia, 19.2%; other infections, 33.4% of cases. The overall incidence of mortality was 22.6% and of mortality due to infections 17.4%. All FUO episodes resolved. Pneumonia was the major cause of death; 60% of recipients who developed pneumonia died, accounting for 90% of deaths attributable to febrile complications. Interstitial pneumonia, occurred rarely, in 3.9% of all febrile episodes. The Cox model showed that the presence of graft-versus-host disease (GVHD) was related to an approximately ninefold increase in the risk of a first episode of FUO (P value .03). The risk of developing pneumonia was fourfold greater in children who received a transplant from a matched unrelated donor or a mismatched family donor (P value .01). Developments in diagnostic tools are needed to diagnose febrile episodes earlier and more precisely with the aim of reducing early mortality after BMT. PMID- 9211539 TI - Transcription of tal-1, a putative oncogene playing an important role in childhood T-ALL, can be shown in normal peripheral blood cells by a highly sensitive RT-PCR assay. AB - Rearrangement of the tal-1 gene is the most frequent clonal marker in childhood T cell acute leukemia. Previously, tal-1 mRNA expression has been observed only in cells of the erythroid, mast cell, and megakaryocytic lineages and in blastic lymphoid cells of normal bone marrow, not in normal lymphocytes or monocytes of the peripheral blood (PB). In this study we addressed the question of tal-1 expression during normal hematopoietic development by performing reverse transcription-polymerase chain reaction (RT-PCR) on RNA from PB cells of 12 healthy donors. Ten of 10 unsorted samples were RT-PCR positive for tal-1 expression. Sorted T cells and monocytes from three donors showed tal-1 RT-PCR products. This is the first direct experimental evidence of tal-1 transcripts in these two normal PB cell types. PMID- 9211541 TI - Thrombocytopenia in neonates born to women with autoimmune thrombocytopenic purpura. AB - We conducted a survey by questionnaire to clarify the actual conditions of neonates born to mothers with autoimmune thrombocytopenic purpura (ATP) in Japan. We investigated 93 pregnancies (1 resulting in twins) in 31 hospitals between 1985 and 1994. Forty-nine of the neonates (52%) had thrombocytopenia (below 150 x 10(9)/L). Nineteen neonates (20%) showed a bleeding tendency, but this was generally mild. In only one neonate (1%) (a case of asymptomatic intracranial hemorrhage, ICH), deep bleeding occurred due to thrombocytopenia. The lowest platelet count of neonates after birth occurred on day 4, not on day 0. There was no correlation between maternal and neonatal platelet counts. However, there was an apparent correlation between the neonatal platelet count on day 0 and the lowest platelet count after birth. Treatment of the mothers with intravenous high dose gamma-globulin and prednisolone did not prevent risk of neonatal thrombocytopenia significantly. PMID- 9211540 TI - Hodgkin's disease in 82 Turkish children diagnosed over a 10-year period: epidemiological, clinical, and histopathologic features and prognosis with prolonged chemotherapy. AB - In this study, 82 Turkish children with Hodgkin's disease (HD) between 1 and 14 years of age and diagnosed over a 10-year period were evaluated retrospectively. More than half of the patients (54%) presented with advanced stages of HD. Mixed cellularity (MC) was the most frequent (56.1%) histopathologic type, which was followed by nodular sclerosing (NS, 18.3%) in frequency. None of the patients received radiotherapy as initial treatment. In 67 children the COPP regimen alone and in 15 the ABVD regimen alternating with COPP were started, to be given as a total of 12 courses. In the patients who presented with stage I-II HD the overall survival (OAS) rate and 5-year event free survival (EFS) rate were 92.3% and 77.8%, respectively. In the patients with advanced disease (stage III-IV) OAS and 5-year EFS were estimated to be 89.5% and 67.4%, respectively. No serious toxicity of chemotherapy was detected during the follow-up. In this group, clinical, epidemiological and histopathologic features of the disease showed a special pattern close to the type I pattern of HD. Regarding the survival rules and occurrence of low toxicity in our patients, results of prolonged chemotherapy alone seem to be encouraging in most of the children with HD. However, the follow up duration is not yet sufficient to declare a clear conclusion related to the late complications. PMID- 9211542 TI - Reversible occlusion shunt for intraventricular chemotherapy in shunt-dependent brain tumor patients. AB - Intraventricular chemotherapy is increasingly used in the treatment of pediatric brain tumors with leptomeningeal seeding. However, some patients are shunt dependent after surgery, probably due to adhesions in the area of surgery. To avoid drug diversion in these patients we connected the reservoir to a reversible occlusion device. Over a 2-year period a shunt value with an on-off device was inserted into the shunt assembly of eight children with various brain tumors with a poor prognosis undergoing intraventricular chemotherapy. All eight patients had tumor cells in the ventricular cerebrospinal fluid (CSF) and/or metastases by magnetic resonance imaging. The number of intraventricular drug applications ranged from 10 to 51. No shunt malfunctions or shunt-related infections occurred. The temporary closure of the shunt after drug delivery was well tolerated. In all six children with tumor cells in the ventricular CSF a negative cytology was achieved over a 3- to 8-week period. PMID- 9211543 TI - Bleomycin and cyclophosphamide toxicity simulating metastatic nodules to the lungs in childhood cancer. AB - Two pediatric oncology patients with Ewing's sarcoma and one with mixed germ cell tumor were treated with drug regimens that included bleomycin or cyclophosphamide. Despite progress to apparently complete remission, all manifested pulmonary nodules on computed tomography during or at the end of treatment. Thoracoscopic biopsy to confirm metastasis revealed instead fibrotic lesions apparently attributable to bleomycin or cyclophosphamide. After cessation of chemotherapy, the pulmonary lesions resolved and all three patients sustained their remissions. The case histories and comments on the diagnosis and management of pulmonary nodules are reviewed. PMID- 9211545 TI - Point-of-care testing--the cost of convenience. PMID- 9211544 TI - Congenital hemangiopericytoma/infantile myofibromatosis: radical surgery versus a conservative "wait and see" approach. AB - Infantile/congenital hemangiopericytoma, although sharing many similar histological features with adult hemangiopericytoma, has a much better prognosis. Nevertheless, most cases described in the literature were pursued by radical surgery with or without adjuvant chemotherapy. We describe a neonate who presented with a huge mass in the right gluteus, 6 x 5 x 4 cm, and a small ventral abdominal mass. The masses were confirmed on biopsy according to light microscopy, immunohistochemistry, and electron microscopy as congenital hemangiopericytoma. They shrank spontaneously within 2 weeks and vanished within 2 months. We present a hypothesis that masses appearing in the neonatal period with this histology and with no life-endangering pressure on vital organs should routinely be dealt with conservatively. PMID- 9211546 TI - Reduced inferior olivary neuron number in early Down syndrome. AB - Counts of total neuron number per section and of neurons per microscopic field of inferior olivary principal nuclei were made on sections from 10 patients with Down syndrome (DS) aged 0.36 to 28 months and seven control (C) patients aged 1 to 29 months. After stereologic and appropriate shrinkage corrections of the count data, the ratios of values for DS/C were calculated. For mean principal olivary nucleus neuron number, DS/C = 0.64; for mean number of neurons per field, DS/C = 0.84; for mean volume of olivary neuronal band per section, DS/C = 0.79; and for mean volume of neuronal band per neuron, DS/C = 1.27. The data are in accord with other data suggesting that (1) numbers of cells in various cell populations, including various areas of the cerebrum, in DS approximate two thirds normal (DS/C approximately 0.67); (2) for the volumes of such cell populations, DS/C = 0.82 normal; and (3) for volumes of individual cells, DS/C = 1.22 normal. The data of the present study suggest that the inferior olivary nuclei in DS are affected in the same way and to a similar degree as other brain areas, with the age distribution and histologic features of the specimens studied suggesting that the reduced olivary principal nucleus number in early Down syndrome results from reduced initial neuron production rather than postnatal neuron loss. PMID- 9211547 TI - Inflammatory cytokine mRNAs in surgical specimens of necrotizing enterocolitis and normal newborn intestine. AB - Coagulation necrosis, inflammation, and hemorrhage are pathologic hallmarks of necrotizing enterocolitis (NEC). Because cytokines are peptides that mediate inflammatory cell recruitment and amplify the immune response, several of the inflammatory cytokines have been implicated in NEC. We hypothesized that mRNA levels for the interrelated cytokines interleukin-1 beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), IL-6, and the neutrophil chemotactic factor IL 8 would be increased in NEC and would be associated with the presence of inflammation. In this study, we determined the relative levels and localization of mRNA for these cytokines in surgical pathology archival intestinal tissue from 29 premature infants with acute NEC and 15 control infants with congenital intestinal malformations using a novel quantitative in situ hybridization technique. Compared with controls, there were higher IL-1 beta mRNA levels in full-thickness sections and higher TNF-alpha mRNA levels in full-thickness and mucosa sections of acute NEC samples, suggesting a potential role for these cytokines in the pathogenesis of local inflammation in NEC. IL-6 and IL-8 mRNA levels were similar in samples of control and acute NEC cases. Analysis of covariance including all subjects showed that the presence of acute inflammation was associated with increased IL-1 beta mRNA levels in mucosa (P = .035) and increased IL-8 in full-thickness sections (P = .005) and mucosa (P = .01). In four of five NEC cases in which intestinal specimens were available from reanastomosis surgery, cytokine mRNA levels decreased to low or undetectable levels. These data suggest that the inflammatory cytokines are involved in neutrophil recruitment and augmentation of the inflammatory response in neonatal intestine. PMID- 9211548 TI - Cell populations in the bone marrow of guinea pig fetuses with intrauterine growth retardation. AB - Bone marrow smears and blood samples were examined in guinea pig fetuses in which intrauterine growth retardation (IUGR) had been induced by uterine artery ligation and compared with those of control (well-grown) fetuses from uterine horns with intact circulation. Differential bone marrow cell counts were obtained from a count of 300 cells per smear and blood samples were assayed for hemoglobin concentration and 2,3-diphosphoglycerate (DPG). Results of blood assays showed no difference in hemoglobin concentration. DPG levels were reduced in the IUGR guinea pigs (P < .05), which could be a consequence of decreased glucose availability or represent an adaptation to reduced oxygen availability. Comparisons of bone marrow counts revealed an increase in total erythrocyte precursors (P < .05) and a decrease in total granulocytic precursors (P < .05) in IUGR fetuses. Within the erythroid lineage there was a significant increase in late (orthochromatic) erythroblasts (P < .005) in the IUGR animals compared with control animals. The granulocytic lineage of the IUGR fetuses showed a significant decrease in mature neutrophils (P < .05) and eosinophilic precursors (P < .05) compared with controls. These data suggest that the hypoxic stress of uterine artery ligation leads to an increase in medullary erythropoiesis. In concert with a previous study that showed a reduction in hepatic erythropoiesis, these data suggest a precocious shift of the anatomic site of erythropoiesis from the liver to the bone marrow under conditions of hypoxia. PMID- 9211549 TI - Opportunistic infections in pediatric HIV infection: a study of 74 autopsy cases from Latin America. The Latin American AIDS Pathology Study Group. AB - The present report describes opportunistic infections found at 74 autopsies of pediatric HIV/AIDS patients performed at several hospitals in Latin American countries. Fungal infections were the most common (53 cases), Candida sp. (39.18%) and Pneumocystis carinii (20.27%) being the most frequently recognized. Other fungal diseases included histoplasmosis, aspergillosis, and cryptococcosis. Viral infections were present in 31 cases, 38.7% being due to cytomegalovirus. Other viruses recognized included herpes simplex and adenovirus. Additional opportunistic infections were due to Mycobacterium avium-intracellulare, toxoplasmosis, and tuberculosis. Nonspecific bacterial bronchopneumonia was present in 11 cases. Cytomegalovirus and P. carinii coinfection was the most common association found. In this series patients died at a younger age (72% at or younger than 1 year old) and there was a slightly higher number of cases of histoplasmosis and brain toxoplasmosis than in other previously published series of infants and children. PMID- 9211550 TI - Malignancy metastatic to the products of conception: a case report with literature review. AB - Breast cancer is rare in adolescent females. Breast cancer metastatic to the products of conception is equally uncommon. We describe a 15-year-old girl who at 30 weeks of gestation was diagnosed with metastatic adenocarcinoma of probable breast origin. The placenta showed extensive intervillous disease. Metastatic disease within the intervillous space indicates hematogenous dissemination of cancer and a poor prognosis for the mother. The infant is almost always free of maternal disease unless there is villous invasion. Hormonal changes or immunotolerance by the mother may be involved in the pathogenesis. All placentas in which maternal malignancy is known or suspected should be examined grossly and microscopically. PMID- 9211551 TI - Hyaline cartilage at porta hepatis in extrahepatic biliary atresia. AB - Hyaline cartilage was found on microscopy of sections of the extrahepatic biliary tree in two infants with extrahepatic biliary atresia (EHBA). Respiratory epithelium was not present, and the cartilage did not seem to block the bile duct lumen. Hyperbilirubinemia was manifest in one infant on the second postnatal day, but clinical courses were otherwise unremarkable. In neither infant was the ductal plate malformation found on light microscopy of liver biopsy specimens, and in neither infant was visceral topography abnormal. Hyaline cartilage at the porta hepatis appears to be a novel finding in EHBA. Its significance remains to be defined. PMID- 9211552 TI - Disseminated calcification with predominant muscle and cerebral involvement in a child with acquired immunodeficiency syndrome: a case report. AB - Calcification, as a feature of the vasculopathy, is a common pathological finding in children with acquired immunodeficiency syndrome. Large and small blood vessels of many organs including brain, heart, lungs, kidneys, liver, and spleen are affected. We report a case with calcification in multiple organs that was most prominent in the cardiac and skeletal muscle cells and the blood vessels of the cerebrum. To our knowledge, calcification of this degree has not been documented previously in a human immunodeficiency virus-infected child. PMID- 9211553 TI - Hepatocellular carcinoma following neonatal hepatitis. AB - Hepatocellular carcinoma is an uncommon malignancy in young children associated with a variety of congenital and acquired conditions. It has been generally held that idiopathic neonatal hepatitis is not an antecedent of hepatocellular neoplasia in childhood. We report a 28-month-old girl in whom a diagnosis of neonatal giant cell hepatitis was confirmed by liver biopsy at 4 months of age who was followed up with serial liver biopsies. Hepatitis B and C virus infection and metabolic abnormalities had been excluded by appropriate testing. There was no history of parenteral nutrition. The morphologic criteria for a diagnosis of cirrhosis were satisfied in a liver biopsy undertaken at 23 months of age. At 28 months a laparotomy was performed because of continuing jaundice and the development of an abdominal mass. Biopsy of the mass revealed a hepatocellular carcinoma. Ploidy studies showed an aneuploid tumor and a hyperdiploid karyotype was confirmed by chromosomal analysis. This case demonstrates by sequential biopsy the progression from neonatal hepatitis to cirrhosis and hepatocellular carcinoma in a young child. PMID- 9211554 TI - Mucosal calcified nodule of the hard palate in an infant: case report and review of the literature. AB - Pathological calcifications of skin manifest as small or large deposits of calcium in the dermis and subcutaneous tissues. One form of these conditions is described as subepidermal calcified nodule seen on the facial skin of young children without any underlying connective tissue disease or any abnormality in calcium or phosphorus metabolism. The oral cavity is rarely affected. Recently, two cases were reported in the oral mucosa and the term "mucosal calcified nodule" was coined for such an entity. We report another case of such a process involving the oral mucosa of a 5-month-old infant who presented with an enlarging lesion at the junction of the hard and soft palate. PMID- 9211555 TI - Lymphangiomatosis of the body wall: a report of two cases associated with chylothorax and fatal outcome. AB - We report on two cases of an unusual but distinctive variant of lymphangiomatosis, presenting at birth with predominantly cutaneous involvement of the body wall and complicated by chylothorax. The lesion manifested clinically as a slowly progressive, diffuse, and fluctuant skin swelling. Eventually, almost the entire trunk became affected. There were no bone lesions. Histologically, the soft tissues were diffusely infiltrated by interconnecting mazelike lymphatic vessels. The two infants died from infection at the age of 23 days and 10 months, respectively. Our experience confirms that premortem histologic diagnosis of lymphangiomatosis is difficult to establish. Awareness of the condition and knowledge of its various clinical presentation forms are essential for proper recognition, assessment of the outcome, and evaluation of new therapeutic measures such as interferon. PMID- 9211556 TI - Skeletal abnormalities in Meckel syndrome. AB - Meckel syndrome is an autosomal recessive condition with a wide phenotypic variation. The most consistent features are cystic kidneys and intrahepatic bile duct anomalies, frequently accompanied by central nervous system (CNS) malformations and polydactyly. Approximately one sixth of all cases also show skeletal anomalies. We present two cases, siblings born to a consanguineous couple, in whom there was a striking curvature and shortening of the long bones in addition to cystic kidneys, CNS abnormalities, and polydactyly. Histological examination of the long bones in the second affected sibling showed mid diaphysial ectopic cartilaginous growth plates differentiating the long bone changes from other skeletal dysplasias with similar radiological features. PMID- 9211557 TI - Lymphangioma of the right kidney in an infant boy. AB - A 9-month-old boy with a primary cystic lymphangioma of the right kidney is reported. The clinical and radiological features favored a malignant tumor. Histology revealed the lesion to be a lymphangioma; the diagnosis was confirmed by immunohistochemistry and chemical analysis of the intracystic fluid. Lymphangiomas of the kidney are rare in adults; they are even rarer in infants and children. Nevertheless, renal lymphangiomas should be considered in the differential diagnosis of multicystic, unilateral renal masses, independent of the age of the patient. PMID- 9211558 TI - Monoamniotic twins delivered liveborn with a forked umbilical cord. AB - Monoamniotic twins are rare and are associated with high intrauterine mortality rates. This case appears to represent the first report of liveborn monoamniotic monochorionic twins delivered with a bifurcated umbilical cord. Pathological and angiographic studies of the placenta demonstrated a marginally inserted two vessel umbilical cord that bifurcated at 8.4 cm from the disk into three-vessel umbilical cords supplying each twin. This probably represents the last opportunity for cleavage of the embryo prior to the formation of conjoined twins. A review of eight prior reports of monoamniotic twins with a single, bifurcating umbilical cord is provided. PMID- 9211559 TI - Major histocompatibility complex class II deficiency needs an early diagnosis: report of a case. AB - Major histocompatibility complex (MHC) class II deficiency is a rare primary immunodeficiency disorder characterized by defects in human leukocyte antigen class II expression, inconsistent expression of human leukocyte class I molecules, and a lack of cellular and humoral immune responses to foreign antigens. Clinical onset occurs early in life with recurrent infections and chronic diarrhea. The prognosis is poor, and mean age at the time of death is 4 years. The only curative treatment is bone marrow transplantation (BMT), which allows the immune system's reconstitution. BMT should be done early in life, because long-term survival seems to depend on the number of previous viral infections. We report the case of an MHC class II deficiency discovered late in a 4-year-old girl by means of immunohistochemistry of small bowel biopsy revealing the absence of MHC class II expression. The child received a BMT twice but died because of a overwhelming viral infection. This case underlines the necessity to explore children presenting with infections and chronic diarrhea in order to find MHC class II deficiency. Usually, diagnosis is performed on cytospins, but when it has been missed clinically, it can be performed by using immunohistochemistry on small bowel biopsies. PMID- 9211560 TI - Right-sided high origin of diaphragm associated with accessory lobe of liver, lobulated right atrial appendage, and ipsilateral phrenic nerve hamartoma: a case report. AB - A case of a rare condition of congenital right anterior high origin of the diaphragm in a stillborn fetus is reported. Associated findings at autopsy were a hornlike subdiaphragmatic intrathoracic accessory lobe of the liver and a lobulated right atrial appendage of the heart. At the superiormost aspect of the malpositioned right anterior diaphragmatic leaf a small phrenic nerve hamartoma was found. The phrenic nerve itself appeared small and not well developed. The phrenic nerve lesion may have been a concomitant or secondary hamartomatous change. Careful clinical and pathological search for concomitant anomalies in diaphragmatic lesions is emphasized. PMID- 9211561 TI - Hepatic heterotopias in the jejunum: a case study over time showing progressive degenerative changes. AB - Multiple foci of heterotopic liver in the jejunum were sequentially discovered in an infant boy at the ages of 1 day, 2 months, and 4 months. This is the second reported case of jejunal heterotopic liver, a rare entity in any site. Progressive histological changes indicative of biliary duct obstruction were observed in the hepatic heterotopias, which demonstrated no connections to the main body of the liver or biliary tree. PMID- 9211562 TI - Pioglitazone: in vitro effects on rat hepatoma cells and in vivo liver hypertrophy in KKAy mice. AB - Pioglitazone increases insulin sensitivity in vivo and in vitro. The effects of this agent on insulin-induced DNA synthesis and hepatic cell growth have not been determined. We examined the ability of pioglitazone to enhance basal and insulin stimulated DNA synthesis in rat H4IIE (H4) hepatoma cells, and to alter liver weight and histology in diabetic KKAy mice. Treatment of H4 cells with increasing concentrations of pioglitazone for 30 h increased basal DNA synthesis 1.6- to 1.8 fold. With pioglitazone pretreatment and submaximal insulin concentrations, DNA synthesis was significantly increased from 2.1-fold (insulin 10(-12) mol/l alone) to 3.9-fold (insulin 10(-12) mol/l + pioglitazone 10(-6) mol/l). At maximal concentrations of insulin, the enhancement of DNA synthesis increased from 7.4 fold (insulin 10(-8) mol/l alone) to 16.2-fold (insulin 10(-8) mol/l + pioglitazone 10(-6) mol/l). Glyburide did not increase basal or insulin stimulated DNA synthesis. In diabetic KKAy mice, serum glucose levels decreased and body weight, liver weight and liver weight as a percentage of body weight increased following pioglitazone treatment. Histological studies demonstrated marked hepatocyte distension. Our findings suggest that pioglitazone acts as an insulin sensitizer in rat hepatoma cells, increasing basal and insulin-stimulated DNA synthesis, and stimulating fat synthesis and liver hypertrophy in diabetic KKAy mice. PMID- 9211563 TI - Dose-response studies of interferon-alpha 2b on liver fibrosis and cholestasis induced by biliary obstruction in rats. AB - Interferons have been utilized widely in chronic liver diseases for their antiviral properties. In addition, there is evidence for their antifibrogenic actions. In this work we studied effects of various doses of interferon-alpha 2b on experimental liver fibrosis and cholestasis induced in the rat by biliary obstruction. Collagen was measured as hepatic hydroxyproline content. Cholestasis was determined by serum alkaline phosphatase and gamma-glutamyltranspeptidase activities and by bilirubin content. Glycogen was measured in the liver. Interestingly, the best effects (antifibrotic and anticholestatic) were observed in the group receiving the lowest dose of interferon. These results suggest that interferon-alpha 2b may be used at low doses, thereby decreasing side effects and costs. PMID- 9211564 TI - Serotonergic receptors modify the voluntary intake of alcohol and morphine but not of cocaine and nicotine by rats. AB - Effects of fluvoxamine, a relatively selective 5-HT uptake inhibitor, and ipsapirone, a relatively selective 5-HT1A agonist, were studied on the initiation and/or maintenance of the voluntary intake of alcohol, morphine, cocaine, and/or nicotine in rats using the two-bottle free-choice method. Fluvoxamine (30 mg/kg/day in the drinking fluid) when given during existing morphine consumption increased the intake of this drug (1 +/- 1 vs. 3 +/- 1 mg/kg/day) but had no effect on alcohol (2 +/- 2 vs. 2 +/- 2 g/kg/day) or cocaine (10 +/- 10 vs. 13 +/- 10 mg/kg/day) intake. Ipsapirone (10 mg/kg/day in the drinking fluid) when given during existing alcohol or morphine consumption decreased the intake of the first (2 +/- 2 vs. 1 +/- 1 g/kg/day) and increased the intake of the second drug (2 +/- 1 vs. 4 +/- 1 mg/kg/day), but had no effect on nicotine (1 +/- 1 vs. 1 +/- 1 mg/kg/day) or cocaine (7 +/- 8 vs. 7 +/- 6 mg/kg/day) intake. Ipsapirone when given before exposure to the above drugs reduced subsequent alcohol (2 +/- 1 vs. 1 +/- 1 g/kg/day) and increased subsequent morphine intake (2 +/- 2 vs. 4 +/- 1 mg/kg/day), but had no effect on the voluntary consumption of cocaine (8 +/- 7 vs. 10 +/- 6 mg/kg/day) and nicotine (1 +/- 1 vs. 1 +/- 1 mg/kg/day). These results suggest: (1) selective stimulation of 5-HT1A receptors reduces alcohol preference, (2) stimulation of all 5-HT receptors has no effect on alcohol intake, indicating the presence of inhibitory receptors, (3) stimulation of the serotonergic system in general stimulates morphine preference, (4) the serotonin system does not affect nicotine or cocaine preference and (5) the serotonergic system is not involved in the voluntary consumption of all, but-only of some drugs/chemicals of abuse. Recognition of these drug/chemical-specific sites in the brain might lead to a better understanding of differences in drug abuse patterns among humans and help in the development of specific drugs for the treatment of selective drug addictions. PMID- 9211566 TI - Protective effect of acetaminophen against acute gastric mucosal lesions induced by ischemia-reperfusion in the rat. AB - We examined the effect of acetaminophen, an analgesic and antipyretic drug, on acute gastric mucosal injury induced by ischemia-reperfusion in rats. Ischemia reperfusion was induced by clamping the celiac artery for 30 min with a small clip. Sixty minutes after reperfusion, the total area of erosions was measured. Acetaminophen (300 and 500 mg/kg) administered intraperitoneally 90 min before the ischemia significantly reduced the total area of erosions. The drug also inhibited the increase in lipid peroxide levels induced by ischemia-reperfusion in the gastric tissue and the increase in lipid peroxidation caused by the hydroxyl radical, OH., in vitro. Gastric prostaglandin E2 (PGE2) contents tended to increase after ischemia-reperfusion in both control and acetaminophen-treated groups. A significant difference in gastric PGE2 contents between control and acetaminophen-treated groups was not observed. The results indicate that acetaminophen may protect the gastric mucosa against ischemia-reperfusion injury, probably by blocking hydroxyl-radical-induced membrane damage. PMID- 9211565 TI - Cinitapride protects against ethanol-induced gastric mucosal injury in rats: role of 5-hydroxytryptamine, prostaglandins and sulfhydryl compounds. AB - This study was designed to determine the gastroprotective properties of cinitapride (CNT), a novel prokinetic benzamide derivative agonist of 5-HT4 and 5 HT1 receptors and 5-HT2 antagonist, on mucosal injury produced by 50% (v/v) ethanol. Results were compared with those for 5-hydroxytryptamine (5-HT: 10 mg kg 1). The possible involvements of gastric mucus secretion, endogenous prostaglandins (PGs) and sulfhydryl compounds (SH) in the protection mediated by CNT were also examined. Intraperitoneal administration of CNT (0.50 and 1 mg kg 1), 30 min before ethanol, significantly prevented gastric ulceration and increased the hexosamine content of gastric mucus. CNT (1 mg kg-1) also produced a significant increase in gastric mucosal levels of PGE2, but did not induce any significant changes in SH values. On the contrary, pretreatment with 5-HT worsened ethanol-induced erosions, however, did not affect gastric mucus secretion, glycoprotein content or PGE2 levels, although the non-protein SH fraction was significantly decreased. The present results demonstrate that the gastroprotective effects of CNT could be partly explained by a complex PG dependent mechanism. We suggest that 5-HT dependent mechanisms through 5-HT2 receptor blockade and 5-HT1 receptor activation could be also involved. PMID- 9211567 TI - Vanidilol: a vanilloid-type vasorelaxant and ocular hypotensive beta-adrenoceptor blocker with partial beta-2-agonist activity. AB - Vanidilol, [4'-(2-hydroxy-3-(tert-butylamino)propoxy)-3'-methoxyphenyl] benzaldehyde, newly synthesized from vanillin, is a vanilloid-type beta adrenoceptor blocker. The beta-adrenoceptor-blocking properties of vanidilol were studied both in vivo and in vitro. Intravenous injection of vanidilol (1.0, 3.0, 5.0 mg/kg) in anesthetized Wistar rats produced a decrease in blood pressure and a dose-dependent bradycardia response. Vanidilol inhibited the tachycardia effects induced by (-)isoproterenol, but had no blocking effect on the arterial pressor responses induced by phenylephrine. In isolated guinea-pig tissues, vanidilol attenuated the (-)isoproterenol-induced positive chronotropic and inotropic effects of the atria and trachea relaxation responses in a concentration-dependent manner. The parallel shift to the right of the concentration-response curve of (-)isoproterenol suggested that the agent was a beta-adrenoceptor competitive antagonist. The apparent pA2 values for vanidilol on the right atria, left atria and trachea were 7.67 +/- 0.03, 7.89 +/- 1.02 and 7.66 +/- 0.15, respectively, denoting that vanidilol was a nonselective beta blocker. The intrinsic sympathomimetic activity of vanidilol and propranolol was determined on isolated atria and trachea from reserpinized guinea pigs. Propranolol caused significantly negative inotropic and chronotropic effects at 10(-6) mol/l or above, whereas vanidilol possessed less cardiodepressant activities than propranolol. In reserpinized tracheal strips, vanidilol produced dose-dependent relaxant responses, but propranolol was ineffective. Preincubating the preparations with ICI 118,551 (0.1-10 nmol/l), a beta 2-adrenoceptor antagonist, significantly shifted the concentration-relaxation curves of vanidilol to a region of higher concentrations. In isolated guinea-pig thoracic aorta, vanidilol (0.1-10 mumol/l) inhibited the phenylephrine (10(-5) mol/l) induced tonic contraction in vascular smooth muscle which was related to the block of calcium influx. In 20% saline-perfused rabbits, vanidilol showed a marked delay in intraocular pressure recovery, demonstrating an ocular hypotensive action. Binding characteristics of vanidilol and propranolol were evaluated in [3H]dihydroalprenolol binding to porcine ventricular membranes. Vanidilol was less potent than propranolol in competing for the beta-adrenoceptor binding sites. On the other hand, vanidilol had a high hydrophilicity in comparison with propranolol. In conclusion, vanidilol exhibited nonselective beta adrenoceptor blocking, vasorelaxant and ocular hypotensive activities, but was devoid of alpha-adrenoceptor blocking and beta 1-agonist activity. Partial beta 2 adrenoceptor agonist activity and inhibitory activity on calcium influx may share in the vasorelaxant activity. PMID- 9211568 TI - Molecular genetic aspects of alcohol metabolism and alcoholism. AB - Recent human genetic studies suggest that a predisposition to alcohol abuse and/or to develop alcoholism may be inherited. Pedigree analysis, linkage, and association studies have helped to detect marker loci and candidate genes that may prove useful in identifying individuals at risk. In particular, molecular genetic research into the causes of alcoholism has drawn attention to the potentially important role of alcohol- and acetaldehyde-metabolizing enzymes, alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH). Functional polymorphisms have been observed at various genes encoding these enzyme proteins, all of which act to alter the rate of synthesis of the toxic metabolite acetaldehyde, or decrease its further oxidation. The occurrence of functional polymorphisms in alcohol-metabolizing enzymes makes them favored candidate genes suitable for further molecular genetic research. A positive selection of such genetic polymorphisms in some populations might act as a protective factor against alcohol abuse and alcohol-related disease outcomes. For example, individuals who show initial sensitivity to alcohol by virtue of their genetically controlled abnormality of ALDH2*2 allele are discouraged from excessive alcohol consumption. On the other hand, persons with the heterozygous ALDH2*2 genotype (ALDH2*1/2*2) are at higher risk for developing alcohol abuse related end-organ damage than those with a homozygous ALDH2*1/2*1 genotype. Moreover, the frequency of C2 allele of cytochrome P45 02E1 was found to be higher in patients with nonfibrotic alcoholic liver disease than in patients with severe hepatic fibrosis or liver cirrhosis. Identification of putative alcoholism vulnerability genes by direct analysis of candidate genes and genetic linkage may therefore help improve approaches to prevention and treatment. PMID- 9211569 TI - Effects of fluvoxamine on depression, anxiety, and other areas of general psychopathology in bulimia nervosa. AB - The efficacy of fluvoxamine in maintaining improvement of general psychopathology (depression, obsessive-compulsive symptoms, anxieties, interpersonal trust, and body perception) was tested in a double-blind placebo-controlled study of 72 patients with bulimia nervosa who were being treated successfully with inpatient behavioral psychotherapy. Over a period of about 15 weeks (2-3 weeks inpatient titration phase, 12 weeks outpatient relapse-prevention phase), fluvoxamine or placebo were given. The relapse-prevention design was used to avoid potential confounding effects of other concomitant treatments. Assessments concerning general psychopathology were made on the basis of expert ratings (CGI, HDRS) and self ratings (HSCL, Eating Disorders Inventory (EDI)-subscales "ineffectiveness," "perfectionism," "maturity fears," "interpersonal distrust," and "interoceptive awareness"). Fluvoxamine had significant effects in preventing relapse as measured on the basis of the Clinical Global Impression (CGI) scale "severity of illness", and a positive trend for relapse preventing effects was observed for the HSCL "general symptomatic index". Further, a relapse preventing effect was observed for the HSCL subscale "obsessive-compulsive symptoms", but not for the EDI subscale "perfectionism". Various dependent variables measuring depression showed no significant relapse-preventing effects of fluvoxamine, but only positive trends. Fluvoxamine had no relapse preventing effects according to our results for dependent variables assessing anxieties, interpersonal trust, and body perception. During a final short (4-week) off-medication phase, no statistically significant effects of discontinuation of medication, but some trends in the expected directions, were observed. PMID- 9211570 TI - Combination therapy using moclobemide with tricyclic and tetracyclic antidepressants to treat therapy-resistant depression. AB - In an open trial, a combination therapy with 300 mg moclobemide was instituted after adjustment to a classical tricyclic or tetracyclic antidepressant in a group of 23 previously therapy-resistant depressive inpatients (pretreatment with two biochemically different antidepressants over a period of at least 5 weeks with sufficient doses). In 13 patients (53.9%) a significant improvement on the Hamilton Depression Scale (24-item version) of at least 50% was achieved. The improvement in the BPRS was also significant. On the basis of the present findings, a combination therapy with the reversible MAO-inhibitor moclobemide represents an efficacious regimen for therapy-resistant depressions, with low side-effects. After this pilot study, further controlled studies are necessary. PMID- 9211571 TI - A double-blind study comparing paroxetine and maprotiline in depressed outpatients. AB - A double-blind multicenter randomized parallel group study comparing paroxetine and maprotiline was carried out in a total of 544 outpatients. Included were patients with varying degrees of severity of depressive symptoms who fulfilled modified RDC criteria for either Minor or Major Depression and showed a HAMD-17 score of > or = 13. No concomitant benzodiazepine treatment was allowed. Duration of treatment was 6 weeks, after an initial wash-out period. Doses were fixed during the first 3 weeks of treatment, patients receiving either 20 mg paroxetine or 100 mg maprotiline daily. An option for dose escalation was provided for insufficient responders after 3 weeks. The weekly assessments comprised rating of the HAMD-17, MADRS, BRMS, RDS, HAMA, CAS, and CGI scales and registration of adverse events by non-leading questions. An intention-to-treat and a completer analysis were performed. Response was defined as a HAMD-17 reduction of > or = 50% or a HAMD-17 score of < or = 9 at the end of the study or at dropout. The treatment groups were comparable according to demographic data. Overall evaluation indicated equieffective and good antidepressant and anxiety-reducing properties for paroxetine and maprotiline. No persistent significant differences between treatment groups were observed on any assessment instrument. There was no difference in the frequency of observed side-effects, but side-effect profiles were markedly different, as maprotiline patients had more anticholinergic and paroxetine patients more SSRI-typical side-effects. PMID- 9211572 TI - Serotonin syndrome after lithium add-on medication to paroxetine. AB - A 59-year-old female patient was hospitalized on account of a depressive episode in the course of a long-standing bipolar disorder. On a combination of lithium (400 mg/day) and paroxetine (30 mg/day) she developed symptoms of shivering, high frequency tremor of the upper and lower limbs, skin flush in the face, agitation, and slight impairment of mental focusing, suggestive of a serotonin syndrome. At this stage serum lithium and paroxetine levels were 0.63 mmol/l, and 693 ng/ml, respectively; the latter was six times higher than the upper concentrations seen in patients on this dosage of the drug. Consequently, the dosage of paroxetine was reduced to 10 mg/day, and lithium was continued. This regimen resulted in a steady-state paroxetine serum level of 390 ng/ml. The patient became symptom-free and the depressive episode attenuated, thus enabling us to discharge the patient. PMID- 9211573 TI - Zolpidem dependence in a patient with former polysubstance abuse. AB - Zolpidem is a non-benzodiazepine hypnotic whose actions are mediated at the central GABA-A receptor complex. It has been assumed that zolpidem has a lower potential for abuse than benzodiazepines. However, there is growing evidence form clinical and animal pharmacology that the potential for dependence on zolpidem should not be underestimated (1). In human laboratory studies, abuse liability seemed to be similar to triazolam (2). Animal models suggest that the drug even had higher reinforcing properties than any benzodiazepine tested (3). In the following, a case of zolpidem abuse and dependence is described. The patient had a history of substance abuse. The observed tolerance-related events and withdrawal symptoms were comparable to those maintained by benzodiazepines. PMID- 9211574 TI - Two cases of deep vein thrombosis associated with a combined paroxetine and zotepine therapy. AB - Thromboembolic events arising as a side-effect of neuroleptic and thymoleptic therapy, although rare, represent serious complications. We report on two patients suffering from an acute deep vein thrombosis after administration of a combined therapy with paroxetine and zotepine. The absence of common risk factors for the development of venous thrombosis in both patients led us to reconsider the potential impact that the drugs administered may have on thrombogenesis. Possible influences of the neuroleptic and thymoleptic therapy on the coagulation system are discussed. Although the clinical observations do not necessarily imply a strict causal relationship between drug administration and thrombosis, there are features of these two cases that should enhance our awareness of the possibility of thrombotic events occurring as a consequence of neuroleptic and thymoleptic therapy. PMID- 9211576 TI - Gender differences in the heritability of seasonal mood change. AB - The study estimated gender differences in the magnitude of genetic and environmental influence in seasonal mood change. The self-report Seasonal Pattern Assessment Questionnaire (SPAQ) was completed by 339 volunteer reared-together twinpairs (187 monozygotic pairs, 152 dizygotic pairs) and analysed using biometric genetic models. The SPAQ yields a global seasonality score (GSS) which is an index of change in sleep patterns, social activities, mood, weight, appetite, and energy level. The GSS was significantly heritable among males and females, estimated to account for 69% and 45% of the total variance, respectively. For the individual symptoms, changes in sleep patterns, social activities, mood, appetite, and energy levels were accounted for primarily by additive genetic effects in both males (median, 45.5%) and females (median, 30.5%). For both sexes, weight changes were not heritable. Sex-by-genotype analyses suggested that the genetic factors influencing female seasonality may not be the same as those influencing male seasonality. PMID- 9211575 TI - A linkage study of schizophrenia to markers within Xp11 near the MAOB gene. AB - A sex chromosome locus for psychosis has been considered on the basis of some sex differences in genetic risk and expression of illness, and an association with X chromosome anomalies. Previous molecular genetic studies produced weak evidence for linkage of schizophrenia to the proximal short arm of the X-chromosome, while some other regions were not ruled out. Here we report an attempt to expand the Xp findings in: (i) a multicenter collaboration focusing on 92 families with a maternal pattern of inheritance (Study I), and (ii) an independent sample of 34 families unselected for parental mode of transmission (Study II). In the multicenter study, a parametric analysis resulted in positive lod scores (highest of 1.97 for dominant and 1.19 for recessive inheritance at a theta of 0.20) for locus DXS7, with scores below 0.50 for other markers in this region (MAOB, DXS228, and ARAF1). Significant allele sharing among affected sibling pairs was present at DXS7. In the second study, positive lod scores were observed at MAOB (highest of 2.16 at a theta of 0.05 for dominant and 1.64 at a theta of 0.00 for recessive models) and ALAS2 (the highest of 1.36 at a theta of 0.05 for a recessive model), with significant allele sharing (P = 0.003 and 0.01, respectively) at these two loci. These five markers are mapped within a small region of Xp11. Thus, although substantial regions of the X-chromosome have been investigated without evidence for linkage being found, a locus predisposing to schizophrenia in the proximal short arm of the X-chromosome is not excluded. PMID- 9211577 TI - Twin closeness and co-twin risk for substance use disorders: assessing the impact of the equal environment assumption. AB - Various environmental variables are hypothesized to operate differentially within identical and fraternal twin pairs. To the extent that these factors are correlated with behavioral outcomes, such as alcohol or drug abuse, traditional twin studies of concordance may be biased. Self-ratings of within-pair emotional closeness, assessed in 169 same-sex twin pairs ascertained through alcohol and drug treatment centers, were used to determine the impact of the twin relationship on concordance for alcohol dependence (N = 130 twin pairs) and other drug abuse and/or dependence (N = 85 twin pairs). In general, identical twin pairs reported significantly closer relationships than fraternal twin pairs, and female twin pairs reported significantly closer relationships than male twin pairs. The data did not indicate an overall effect of closeness on co-twin risk for alcohol dependence. In contrast, closeness was significantly related to co twin risk for other drug abuse and/or dependence. However, the MZ/DZ concordance difference for other drug abuse and/or dependence remained significant when the effects of within-pair closeness were controlled. Thus, the initial zygosity and sex differences in concordance for substance use disorders cannot be explained solely by differences in twin relationship due to closeness as assessed in this study. PMID- 9211578 TI - Beneficial effects of thiamine on recognition memory and P300 in abstinent cocaine-dependent patients. AB - The present study evaluated the effects of thiamine vs. placebo on memory task performance and event-related electroencephalographic potentials in eight abstinent cocaine-dependent patients. Patients orally ingested 5 g of thiamine and 5 g of a lactose placebo on two separate days scheduled approximately 1 week apart. The order of administration was randomized. Double-blind procedures were followed. Approximately 3 h after ingesting the capsules, patients completed Sternberg's (1975) memory scanning task during which performance and event related potentials (P300) were recorded simultaneously. Thiamine was found to significantly improve recognition accuracy and P300 amplitude, at the midline parietal (Pz) electrode. The improvement was most reliable under conditions of increased memory load. These preliminary findings justify a further examination of the relation between thiamine's hypothesized effects on central nervous system cholinergic function, and the direct and indirect effects of cocaine abuse. PMID- 9211579 TI - Pain assessment in self-injurious patients with borderline personality disorder using signal detection theory. AB - Signal detection theory measures of thermal responsivity were examined to determine whether differences in reported pain experienced during self-injurious behavior in female patients with borderline personality disorder (BPD) are explained by neurosensory factors and/or attitudinal factors (response bias). Female patients with BPD who do not experience pain during self-injury (BPD-NP group) were found to discriminate more poorly between noxious thermal stimuli of similar intensity, low P(A), than female patients with BPD who experience pain during self-injury (BPD-P group), female patients with BPD who do not have a history of self-injury (BPD-C group), and age-matched normal women. The BPD-NP group also had a higher response criterion, B (more stoical) than the BPD-C group. These findings suggest that 'analgesia' during self-injury in patients with BPD is related to both neurosensory and attitudinal/psychological abnormalities. PMID- 9211580 TI - The Family Attitude Scale: reliability and validity of a new scale for measuring the emotional climate of families. AB - Research on outcomes from psychiatric disorders has highlighted the importance of expressed emotion (EE), but its cost-effective measurement remains a challenge. This article describes development of the Family Attitude Scale (FAS), a 30-item instrument that can be completed by any informant. Its psychometric characteristics are reported in parents of undergraduate students and in 70 families with a schizophrenic member. The total FAS had high internal consistency in all samples, and reports of angry behaviour in FAS items showed acceptable inter-rater agreement. The FAS was associated with the reported anger, anger expression and anxiety of respondents. Substantial associations between the parents' FAS and the anger and anger expression of students was also observed. Parents of schizophrenic patients had higher FAS scores than parents of students, and the FAS was higher if disorder duration was longer or patient functioning was poorer. Hostility, high criticism and low warmth on the Camberwell Family Interview (CFI) were associated with a more negative FAS. The highest FAS in the family was a good predictor of a highly critical environment on the CFI. The FAS is a reliable and valid indicator of relationship stress and expressed anger that has wide applicability. PMID- 9211581 TI - Cultural imagination and individual creativity. New directions in psychoanalytic anthropology. Introduction. PMID- 9211582 TI - Discussion: the heritage of imagination. PMID- 9211583 TI - Dramatization: how dream work shapes culture. PMID- 9211584 TI - Symbols, dreams, and self-reflection: some cases from African suriname in Europe. PMID- 9211585 TI - Male birth-giving in the cultural imagination of the Sambia. PMID- 9211586 TI - The imaginative use of religious symbols in subjective experiences of anorexia nervosa. AB - Clinicians working with contemporary women with anorexia nervosa have commented on the ascetic component in anorexia, meaning their self-denial, heightened morality, opposition between body and spirit, asexuality, and denial of bodily death (Mogul, 1980; Palazzoli, 1978; Rampling, 1985; Sabom, 1985; Turner, 1984). While these clinicians have commented on the asceticism in contemporary anorexia nervosa, they have little to say about the role of culture in subjective experiences of this asceticism. As we have seen, Jane and Margaret used notions of asceticism about food and the body that are a part of their religious beliefs to create a personal meaning system through which they expressed their self starvation. These cases, while supporting clinical studies that point to an ascetic component in modern anorexia, go further to suggest that in some cases, this asceticism may be encoded in religion. Religious anorectics like Jane and Margaret challenge models of anorexia nervosa that understand the condition exclusively in terms of cultural foci on "dieting" and secular ideals of beauty and bodily thinness for women (Bemporad, Hoffman, & Herzog, 1989; Chernin, 1985; Garner et al., 1980; Orbach, 1986; Rost, Newhaus, & Florian, 1982). They also suggest a continuing persistence into the twentieth century of an association between religiosity and self-starvation noted by historians during the early Christian, medieval, and late-Victorian periods in the West (Bell, 1985; Brown, 1988; Brumberg, 1985, 1988; Bynum, 1987). The above discussion points to the new directions in psychological anthropology which challenge a strict and opposing dichotomy between the conscious and unconscious, between culture (seen as "public") and the individual mind (seen as "private" and idiosyncratic). Obeyesekere's concept of "the work of culture," (Obeyesekere, 1990) and Stephen's concept of the "autonomous imagination" are especially useful in understanding how persons like Jane and Margaret use in imaginative ways cultural symbols, such as notions of asceticism about food and the body that are a part of religion, to give meaning to their personal concerns with growth, separation, and sexuality. We saw how Jane and Margaret transform cultural symbols and language to express their starvation and deep anxieties. These cases lend support to views that culture and religion, as symbolic systems, have underpinnings in deep motivation (Obeyesekere, 1981, 1990; Spiro, 1965, 1987). They also suggest that the relations between culture and the individual mind (and between culture and "illness," between "normal" and "abnormal") must be viewed as a moving continuum, with culture constantly worked and reworked by the individual imagination in innovative and creative ways. PMID- 9211587 TI - Spiritual healing and human development in the Native American church: toward a cultural psychiatry of peyote. PMID- 9211588 TI - Emotion of Hazukashii in reunion situations for the Japanese. PMID- 9211589 TI - Dreams, ghosts, tales: parintintin imagination. PMID- 9211590 TI - From the sway of the pleasure principle: ghost of a tiger. PMID- 9211592 TI - [Choice of method for statistical analysis of quantitative data obtained from toxicological studies--toxicological data]. AB - We compared the usefulness of t-test and parametric and rank-sum tests in the statistical analysis of significant differences in the so-called "decision tree" for the quantitative data obtained from toxicity studies. The Dunnett's multiple comparison test had lower analytic power than the t-test when one of the groups showed a marked difference in variance. The Dunnett's test was less efficient with the increase in the number of groups. If one group showed a decrease in the number of animals, this test was less efficient than parametric tests, because the rank-sum tests should be chosen. The rank-sum test is required occasionally to attach the asterisks of significant difference to the mean +/- SD even in showing the same mean values. The nonparametric Dunnett's test could not be used for analysis of significant differences when the mean value for the control and treated groups showed big differences. The nonparametric Dunnett's and parametric Scheffe tests were not as efficient as the other parametric tests probably because of the vague evaluation or overlooking the effect of the test substance. PMID- 9211593 TI - Effects on complement, granulocytes and platelets of a leukocyte-depletion filter during in vitro extracorporeal circulation. AB - In an in vitro study, extracorporeal circuits equipped with either a leukocyte depleting filter (n = 5) or a standard arterial-line filter (n = 5) were perfused for 120 minutes with fresh human whole blood. Leukocyte activation, leukocyte and platelet counts and complement activation were studied. Significant reduction of leukocyte and platelet counts and significant activation of leukocytes and of platelets were found in both groups, but without significant intergroup difference for any parameter after 120 minutes of perfusion. The leukocyte depleting filters, however, were somewhat more effective in removing leukocytes during the initial 30 minutes of circulation. PMID- 9211591 TI - [Changes in porphyrin metabolism of mice given beryllium and/or zinc]. AB - Beryllium chloride and/or zinc chloride were intraperitoneally injected into mice. The amount of beryllium (Be) injected corresponded to 1/10th of the LD50 dose intravenously administered. The amount of zinc (Zn) injected was the same as Be. The changes in porphyrin metabolism of the mice were studied. Delta aminolevulinic acid dehydratase (ALA-D) activities in the blood were found to increase significantly in Zn and BeZn groups when compared to the control level. The blood porphobilinogen deaminase (PBG-D) activity in the Zn group was slightly less than that in the controls. The ALA-D and PBG-D activities in liver were higher in the Be and BeZn groups than in the controls. The splenic ALA-D activities were significantly higher in the Zn and BeZn groups than in the control and Be groups. The splenic PBG-D activities were markedly higher in the Be and/or Zn groups than in the controls. An increase in ALA-D activities in the blood and spleen was observed in the BeZn group, together with an increase in ALA D activities caused by Zn administration. Furthermore, the increase in PBG-D activities in liver and spleen was observed in the Be and/or Zn groups. The results suggested that chemical similarity between Be and Zn brought about these phenomena. PMID- 9211594 TI - Surgical treatment of endobronchial tuberculosis. AB - The incidence of tuberculosis remains fairly high in some developing countries. Endobronchial tuberculosis may cause bronchostenosis, with potentially severe respiratory symptoms, atelectasis and secondary pneumonitis. Thirty-two surgically treated cases of tuberculous bronchostenosis (33 operations) are presented. In 13 cases-segmental resection or lobectomy was performed with bronchoplastic procedures. Anastomotic stenosis necessitated pneumonectomy 5 years later in one of the 13 and one patient had wound infection. Nineteen patients underwent pulmonary resection without bronchoplasty. Apart from the patient with anastomotic stenosis, all 32 were symptom-free in the follow-up period. Forced expiratory volume was significantly improved in the ten tested patients with bronchoplasty. The results suggest that surgical treatment is safe for endobronchial tuberculosis with poor response to specific chemotherapy. In addition to checking progression of the disease, bronchoplasty helps to preserve lung function. Appropriate chemotherapy should be given for 9-12 months perioperatively to prevent recurrence and restenosis. PMID- 9211595 TI - Haemodynamic and metabolic effects of gallopamil as additive to calcium containing and calcium-free cardioplegic solutions in mature pig hearts. AB - Myocardial haemodynamic and metabolic effects of the calcium-channel blocker gallopamil as additive to calcium-containing (St Thomas Hospital, STH) and calcium-free (Bretschneider procaine-containing, BRT) crystalloid cardioplegic solutions were evaluated. Adult pig hearts (weight 0.033 kg) were randomized to four groups and perfused with 1 litre of cold (4 degrees C) cardioplegic solution; group A: BRT without gallopamil, n = 9, group B: BRT with gallopamil (0.4 microM), n = 8, group C: gallopamil-free STH, n = 8, and group D: STH with gallopamil (0.4 microM), n = 8. After storage at 4 degrees C for 6 hours the hearts were reperfused with blood/Ringer solution in a modified Langendorff model for 60 min. Developed left ventricular pressure, rate-pressure product and +dP/dt were lower in gallopamil-treated hearts during reperfusion (p < 0.05), as were oxygen extraction and oxygen uptake (p < 0.05) and lactate release (p < 0.05). Myocardial blood flow was greater in gallopamil-treated hearts (p < 0.05). In hearts comparable in size and anatomy to the human heart, gallopamil added to both cardioplegic solutions reduced cardiac function and oxygen uptake despite increased myocardial blood flow. The findings suggest reduced myocardial protection after addition of gallopamil to cardioplegic solutions. PMID- 9211596 TI - Expression of adhesion and activation molecules on lymphocytes during open-heart surgery with cardiopulmonary bypass. AB - Open-heart surgery with cardiopulmonary bypass (CPB) and abdominal surgery are associated with lymphocytopenia. We measured a panel of adhesion and activation molecules on lymphocytes to clarify possible association of CPB with increased expression of these molecules. Eight patients undergoing open-heart surgery and eight with abdominal surgery were studied. The adhesion molecules CD11a/CD18 (LFA 1_, CD11c/CD18 and CD44 and the activation molecules CD25, CD69, CD71 and MHCII were measured, using monoclonal antibodies and flow cytometry. Lymphocytopenia was observed during CPB and for some hours after both open-heart and abdominal surgery. The proportion of CD11a/CD18-positive lymphocytes rose from 67.6 +/- 8% to 86.4 +/- 3% after aortic declamping (p < 0.05). The expression of activation molecules CD25, CD69 and CD71 was unchanged during and after open-heart as well as abdominal operations. Thus CPB was associated with increased expression of the adhesion molecule CD11a/CD18 on lymphocytes, while the expression of activation molecules on lymphocytes was unchanged. PMID- 9211597 TI - Traumatic oesophageal perforation. AB - Sixteen patients were treated for traumatic oesophageal perforation (13 cervical, 3 thoracic) over a 16-year period. In 14 cases the trauma was penetrating. The median delay from injury to treatment was 32 hours and the mean period of hospitalization was 26 days. The treatment procedures were two-layer primary closure with or without drainage, drainage alone and near-total oesophageal exclusion with cervical T-tube oesophagostomy. Postoperative complications were cervical oesophageal leak in two patients and tracheo-oesophageal fistula and oesophageal stenosis, each in one case. Of the eight patients treated within 24 hours of perforation, two died, and of the eight treated later, four died (overall mortality 37.5%). The heightened mortality after delayed diagnosis illustrates the prognostic importance of a high index of suspicion. To prevent leakage, buttressing with viable tissue following primary closure can be useful, especially after delayed diagnosis. Because of the continuing controversy concerning management of late-diagnosed oesophageal perforation, individualized treatment is widely advocated. PMID- 9211598 TI - Retrograde internal mammary artery as coronary bypass. A prospective study with postoperative angiographic evaluation. AB - Although the internal mammary artery (IMA) as a coronary graft offers better long term patency than the saphenous vein, a factor limiting its use has been the length of the artery's pedicle. In an attempt to overcome this limitation, we evaluated the use of retrograde right IMA in a prospective study. In ten patients scheduled for routine coronary artery bypass surgery, bilateral IMA grafting was used, the left IMA in routine fashion, but the right IMA dissected from the level of the first rib, cut there and placed as an inverted graft. Three months postoperatively the patients were clinically evaluated with stress exercise test (n = 10) and coronary angiography (n = 9). No patient had recurrence of angina. Angiography revealed patency of the retrograde right IMA graft in six of nine patients. On the basis of these data we do not recommend routine use of retrograde IMA. PMID- 9211599 TI - Bland-White-Garland syndrome in an adult. Case report and review of diagnostic and predictive strategies. AB - In a 27-year-old man with Bland-White-Garland syndrome (anomalous origin of the left coronary artery from the pulmonary artery), comparison was made between conventional diagnostic techniques and radionuclide imaging for selection of surgical procedure and evaluating the outcome. Dynamic 99mTc imaging exactly located the left coronary artery orifice, which was not seen on angiography, thereby determining the surgical approach, and 123I study revealed that, despite absence of symptoms, the adrenergic activity of the heart was globally diminished, with limited response to revascularization. PMID- 9211601 TI - False left ventricular aneurysm with ventriculo-bronchial fistula and massive haemoptysis. AB - An intravenous drug user presented with bacteraemia and massive hemoptysis 10 years after a penetrating cardiac injury. He was found to have false left ventricular aneurysm with ventriculo-bronchial fistula. The clinical course suggests that the aneurysm became infected and that the inflammatory process weakened the aneurysmal sac and led to the development of fistula. Prompt recognition of the aneurysm with appropriate surgical repair resulted in a successful outcome. PMID- 9211600 TI - One-stage operation for treatment after delayed diagnosis of thoracic esophageal perforation. AB - Perforation of the thoracic esophagus can be fatal unless diagnosed promptly and treated effectively. The high mortality with delayed treatment is principally due to the inability of effectively closing the perforation and preventing the leakage. We operated one patient with a delayed diagnosis of thoracic esophageal perforation developed after a rigid esophagoscopic procedure. The perforation was closed with primary sutures and reinforced with a intercostal muscle flap wrap. Radical decortication and wide mediastinal and pleural toilet were also done. Total parenteral nutrition was begun and antibiotics were administered according to the results of cultures. Esophagography and esophagoscopy performed 10 days after the operation showed a well healed esophagus without stenosis or leakage. We conclude that primary closure of the perforation and muscle flap wrap can provide a one-stage operation with good results for repair of thoracic esophageal perforations which are not diagnosed on time. PMID- 9211603 TI - Tendons--a source of major concern in competitive and recreational athletes. PMID- 9211602 TI - Solitary amyloid nodule in the lung. AB - A rare case of solitary nodular amyloidosis of the lung is presented. As the nodule was clinically observed to enlarge, neoplastic disease was strongly suspected. Histologic study of the resected specimen, however, revealed an amyloid nodule. Three years postoperatively there has been no recurrence. PMID- 9211604 TI - Structure and metabolism of tendons. AB - A tendon forms an integral part of musculotendinous unit. It transmits the tensions generated in muscles to bone. Tendons are stronger than muscles and are able to withstand larger forces. Tendons are subject to both tensile and high compressive forces. PMID- 9211605 TI - Function and biomechanics of tendons. AB - Tendon is a highly organized connective tissue joining muscle to bone, capable of resisting high tensile forces while transmitting forces from muscle to bone. The dense, regularly arranged collagenous tissue is made up of fibers, cells of various shapes and ground substance. The mechanical and physiological characteristics of collagen (nearly 85% of the dry weight of tendon) dictate the qualities of tendon. In addition, tendon is flexible so that it can bend at joints, as well as acting as a damping tissue to absorb shock and limit potential damage to muscle (1). Tendon also shows a degree of extensibility. If the strain used to stretch a tendon could be recovered, a beneficial elastic effect would be achieved. Muscles lengthen and shorten in a cyclical manner. During the lengthening period, elastic energy can be stored and used as elastic recoil. For example, the Achilles tendon is stretched late in the stance phase as the triceps surae muscles contract and the ankle dorsiflexes. Prior to plantarflexion, muscle activation ceases and stored energy helps to initiate planter flexion. PMID- 9211606 TI - Effects of training, immobilization and remobilization on tendons. AB - Since a tendon is a living tissue, it is not a surprise that tendon shows the capacity to adapt its structure and mechanical properties to the functional demands of the entire muscle-tendon unit. However, compared with muscle, the experimental knowledge of the effects of strength or endurance-type training on tendon tissue is scarce and clinical human experiments are completely lacking (1). Research should, however, be able to improve the true understanding of the biomechanical, functional, morphological and biochemical changes that occur in tendons due to training and physical activity, since understanding of the basic physiology of a tissue is the key to understanding its pathological processes (1, 2). Compared with muscle tissue, the metabolic turnover of tendon tissue is many times slower due to poorer vascularity and circulation (1, 3). The adaptive responses of tendons to training are therefore also slower than those in muscles, but they may finally be considerable if the time frame is long enough (3, 4). PMID- 9211607 TI - The aging tendon. AB - After maturation tendons undergo many biochemical, cellular, mechanical and pathological changes that bring about a general decline in the structure and function of the tendon. This decline in the aging tendon is characterized by a reduced ability to adapt to environmental stress and loss of tissue homeostasis. The tendon's adaptability to these changes will decide the rate and the success of treatment of a tendon injury. This review examines these changes and also looks at how we can curtail their progression through exercise and lifestyle modification. PMID- 9211608 TI - Etiology and pathophysiology of chronic tendon disorders in sports. AB - In sports medicine, a chronic overuse injury is defined as a long-standing or recurring orthopedic problem and pain in the musculoskeletal system, which started during exertion due to repetitive tissue microtrauma (1). Repetitive microtrauma, which is basically repeated exposure of the musculoskeletal tissue to low-magnitude forces, results in injury at the microscopic level, and no single acute trauma is normally involved in the pathogenesis of an overuse injury. In chronic tendon disorders, 'overuse' implies that the tendon has been strained repeatedly to 4-8% strain until unable to endure further tension, whereupon injury occurs (2). The structure of the tendon is disrupted micro- or macroscopically by this repetitive strain, i.e. collagen fibrers begin to slide past one another, causing break-age of their cross-linked structure, and denaturate; inflammation, edema and pain result. Thus, tendinitis, peritendinitis, tenosynovitis, insertion tendinitis, tendinous bursitis or apophysitis is the earliest clinically recognizable manifestation of overuse tendon injury (3). PMID- 9211609 TI - Histopathological findings in chronic tendon disorders. AB - Tendon injuries and other tendon disorders represent a common diagnostic and therapeutic challenge in sports medicine, resulting in chronic and long-lasting problems. Tissue degeneration is a common finding in many sports-related tendon complaints. In the great majority of spontaneous tendon ruptures, chronic degenerative changes are seen at the rupture site of the tendon (1). Systemic diseases and diseases specifically deteriorating the normal structure of the tendon (i.e. foreign bodies, and metabolic, inherited and infectious tendon diseases) are only rarely the cause of tendon pathology. Inherited diseases, such as various hereditary diseases with disturbed collagen metabolism and characteristic pathological structural alterations (Ehlers-Danlos syndrome, Marfani syndrome, homocystinuria (ochronosis)), represent approximately 1% of the causes of chronic tendon complaints (2), whereas foreign bodies are somewhat more common and are found in less than 10% of all chronic tendon problems (1). Rheumatoid arthritis and sarcoidosis are typical systemic diseases that cause chronic inflammation in tendon and peritendinous tissues. Altogether, these 'specific' disorders represented less than 2% of the pathological alterations found in the histological analysis of more than 1000 spontaneously ruptured tendons (1, 3, 4). In this material, degenerative changes were seen in a great majority of the tendons, indicating that a spontaneous tendon rupture is a typical clinical end-state manifestation of a degenerative process in the tendon tissue. The role of overuse in the pathogenesis of chronic tendon injuries and disorders is not completely understood. It has been speculated that when tendon is overused it becomes fatigued and loses its basal reparative ability, the repetitive microtraumatic processes thus overwhelming the ability of the tendon cells to repair the fiber damage. The intensive repetitive activity, which often is eccentric by nature, may lead to cumulative microtrauma which further weakens the collagen cross-linking, non-collagenous matrix, and vascular elements of the tendon. Overuse has also been speculated to cause chronic tendon problems, by disturbing the micro- and macrovasculature of the tendon and resulting in insufficiency in the local blood circulation. Decreased blood flow simultaneous with an increased activity may result in local tissue hypoxia, impaired nutrition and energy metabolism, and together these factors are likely to play an important role in the sequence of events leading to tendon degeneration (4). A sedentary lifestyle has been proposed as a main reason for poor basal circulation of the tendon, and presumably is at least partly responsible for the high number of tendon problems in people with a sedentary lifestyle who occasionally take part in high physical activity sports events. PMID- 9211610 TI - Diagnosis and treatment of chronic tendon disorders in sports. AB - Sports and physical activity are becoming more important and more emphasized in the lives of the average person as the health benefits of maintaining an active lifestyle are recognized. In the past most people were primarily active in sports during their time in school. The trend is for more people to continue vigorous activity through middle age and beyond. In addition, as high level athletes continue to reach higher levels of performance more amateur athletes attempt to reach similar levels of intensity, which they may not be able to handle with their level or method of training. This has led to an increase in overuse injuries and chronic tendon injuries. It has been estimated that overuse type injuries account for 30-50% of sports injuries (1). PMID- 9211611 TI - Etiology and pathophysiology of tendon ruptures in sports. AB - Of all spontaneous tendon ruptures, complete Achilles tendon tears are most closely associated with sports activities (1-3). Schonbauer (3) reported that 75% of all ruptures of the Achilles tendon are related to sports. In Plecko & Passl (2) the number was 60%. In our material of 430 cases, the number of sports related Achilles ruptures was very similar (62%), while only 2% of ruptures of other tendons were sports-related (P < 0.001) (1). Also, the majority of Achilles reruptures occurred in sports. The ruptures occurred most often in soccer (34%), track and field (16%) and basketball (14%). The distribution of Achilles ruptures according to different sports varies considerably from country to country, according to the national sport traditions. For example, in northern and middle Europe, soccer, tennis, track and field, indoor ball games, downhill skiing, and gymnastics are the most common; and in North America, football, basketball, baseball, tennis and downhill skiing dominate the statistics (1, 2, 4). In sports, some Achilles ruptures are not spontaneous or degeneration-induced but may occur as a consequence of the remarkably high forces that are involved in the performance (2). Ruptures in the high jump or triple jump are good examples. In such cases, failure in the neuromuscular protective mechanisms due to fatigue or disturbed co-ordination can frequently be found. The spontaneous complete rupture of the supraspinatus tendon of the rotator cuff does not occur very frequently in sports. Those sports that include high-energy throwing movements, such as American and Finnish baseball, American football, rugby and discuss and javelin throwing, may, however, produce this injury. Partial tears and inflammations of the rotator cuff complex are much more frequent in throwing sports. The complete rupture of the proximal long head of the biceps brachii tendon is rare among competitive and recreational athletes. In our material, under 2% of these ruptures were associated with sports activities (5). The rupture (avulsion) of the distal tendon of the biceps muscle is rare. In sports, gymnastics, body building and weight lifting have been said to be able to produce this injury (6). In general, complete ruptures of the quadriceps tendon and the patellar tendon occur most often in older individuals. In our study, the mean age of these patients was 65 years (5). However, these injuries do also occur in younger age groups, especially in athletes. In athletes, the rupture most frequently occurs in high-power sports events, such as high jump, basketball and weight lifting, at the age of 15-30 years. A chronic-patellar apicitis (jumper's knee) may predispose rupture of the tendon (7). As is the case with the rotator cuff complex, overuse inflammation and partial tears of the quadriceps and patellar tendons are one of the most characteristic athletic injuries. Complete spontaneous ruptures of other tendons in sports are rare, although the literature does provide case studies from almost every tendon the human body possesses (8 18). PMID- 9211612 TI - Histopathological findings in spontaneous tendon ruptures. AB - A spontaneous rupture of a tendon may be defined as a rupture that occurs during movement and activity, that should not and usually does not damage the involved musculotendinous units (1). Spontaneous tendon ruptures were uncommon before the 1950s. Bohler found only 25 Achilles tendon ruptures in Wien between 1925 and 1948 (2). Mosender & Klatnek treated 20 Achilles tendon ruptures between 1953 and 1956, but 105 ruptures between 1964 and 1967 (3). Lawrence et al. found only 31 Achilles tendon ruptures in Boston during a period of 55 years (1900-1954) (4). During the recent decades tendon ruptures have, however, become relatively common in developed countries, especially in Europe and North America. A high incidence of tendon ruptures has been reported in Austria, Denmark, Finland, Germany. Hungary, Sweden, Switzerland and the USA; somewhat lower incidences have been reported in Canada, France, Great Britain and Spain. On the other hand, Greece, Japan, the Netherlands and Portugal have reported a clearly lower incidence. Interestingly, Achilles tendon ruptures are a rarity in developing countries, especially in Africa and East-Asia (5). In many developed countries, the increases in the rupture incidence have been dramatic. In the National Institute of Traumatology in Budapest, Hungary, the number of patients with an Achilles tendon rupture increased 285% in men and 500% in women between two successive 7 year periods, 1972-1978 and 1979-1985 (5). PMID- 9211613 TI - Diagnosis, treatment and rehabilitation principles in complete tendon ruptures in sports. AB - Complete tendon ruptures may occur in any tendon subjected to athletic stress. This article discusses diagnostic, treatment and rehabilitation principles that are common to all tendon ruptures. The most common tendon ruptures are then discussed more specifically, presenting pertinent diagnostic tests, treatment considerations and principles of rehabilitation. PMID- 9211614 TI - Work while receiving disability insurance benefits: additional findings from the New Beneficiary Followup Survey. AB - From the foregoing analyses, the following picture emerges about persons who work after award of DI benefits: Almost one-quarter of the sample population attempted to reenter the labor force in the 10-year NBS-NBF period. The higher the level of education, the greater the proportion of persons who worked. Younger beneficiaries were more likely to work than older beneficiaries. About half of the beneficiaries who worked did so on a full-time (40-hour-or-more per week) basis. Most beneficiaries worked because of financial need. The profile of reasons for working did not vary across demographic groups and aspects of the first job held. Most beneficiaries began working without attributing this decision to an improvement in their health. Individuals pursued different methods of job search. No single approach emerged as the most successful. Job search modes did not vary for different groups and different jobs. Four activities were most likely to lead to job offers: persons checking where they had worked before, asking a friend, answering an ad, and following up a vocational rehabilitation lead. These findings were not conclusive because small numbers of persons engaged in these activities. Thirty percent of DI workers returned to their preentitlement employer. The beneficiaries' first postentitlement jobs had less exertion, fewer hours, and lower pay than did their job held prior to award. The likelihood of working was the same across a broad range of disabling health conditions. In terms of work return policy, formal work return programs aimed at young beneficiaries and those with higher levels of educational attainment would produce the greatest number of job placements. It appears that no targeting of programs is necessary along gender lines. The anomalous finding of an absence of the relationship between improvement in health and labor-force reentry requires further investigation. Any followup in this area of inquiry should plan to have the data collected close to the time of postentitlement job entry. PMID- 9211615 TI - Living arrangements of SSI recipients. AB - This article updates one that appeared in the Bulletin in July 1990. It describes living arrangements of persons receiving payments under the Supplemental Security Income (SSI) program from October 1994 through September 1995. The data were taken from the Quality Assurance review conducted by the Social Security Administration (SSA). This procedure is used by SSA to determine the frequency and causes of incorrect determinations of eligibility and payment amounts. It is difficult to describe the living arrangement for the "typical" recipient. Nevertheless, some interesting patterns emerge in an analysis of the data. About 59 percent (owners and renters combined) of the 6.3 million SSI recipients lived in their own households. Approximately 32 percent of them shared a living arrangement with someone else and about 5 percent of the recipients lived in an institution. Of those SSI recipients living in households, about 36 percent lived alone. Less than 13 percent lived with only their spouses or with only their spouses and minor children. Approximately 11 percent of those in households were child recipients living with parents. An additional 15 percent of the SSI recipients lived in households with only other related adults (other than a spouse or parents). PMID- 9211616 TI - Case management at work for SSA disability beneficiaries: process results of the Project NetWork return-to-work demonstration. AB - This article presents the results of the process analysis of the evaluation of the Project NetWork demonstration, a Federal demonstration undertaken by the Social Security Administration (SSA) in 1991 to test alternative methods of providing rehabilitation and employment services to SSA's Disability Insurance beneficiaries and Supplemental Security Income disabled and blind applicants and recipients. The major findings are: (1) from an operational standpoint, it is feasible to expand access to vocational rehabilitation (VR) services to a broad spectrum of SSA beneficiaries, and (2) roughly similar results are achieved, in terms of client intake and provision of services, when case management services are provided by SSA staff, contracted out to State VR agencies, or contracted with private VR providers. Later evaluation reports will trace demonstration impacts on earnings and disability benefits and report the overall benefits and costs of return-to-work services for this population. PMID- 9211617 TI - Ab initio calculation of 1H and 13C NMR shielding constants in solid acetylene. AB - The gauge-independent atomic orbital (GIAO) method has been used within the coupled Hartree-Fock (CHF) approximation to compute 1H and 13C NMR shielding constants for solid acetylene. As the amount of surrounding crystal lattice is increased, the shielding anisotropy of the carbon nuclei decreases and that of the protons increases. The influence of intermolecular interactions on the 13C shielding constant is non-additive. The GIAO approach at the HF level is sufficiently sensitive to differentiate between the two polymorphic forms of acetylene. PMID- 9211618 TI - Effect of 1H-decoupling in two-dimensional multiple-quantum MAS NMR spectroscopy of 23Na in a hydrous layered silicate. AB - In order to check the efficiency of high-power 1H decoupling when used with the two-dimensional multiple-quantum MAS method recently proposed to obtain isotropic spectra from quadrupolar nuclei, we applied this new technique to 23Na NMR for a hydrous layered silicate compound, makatite, with and without 1H decoupling. A remarkable improvement of the spectral resolution in the isotropic dimension was observed by the decoupling, showing that it is effective in the 2D MQ-MAS method and suggesting that proton decoupling should be generally applied in 23Na experiments concerning these kinds of compound. PMID- 9211619 TI - Frequency- and phase-modulated heteronuclear decoupling in rotating solids. AB - The mechanism of heteronuclear dipolar decoupling by the TPPM sequences, proposed by Bennett et al. (J. Chem. Phys. 103 (1995) 6951) is investigated by comparison with a modified pulse sequence, called TPFM, that uses frequency instead of phase modulation. By combining frequency and phase modulations, circularly modulated sequences are designed. The fact that only the left-handed modulation sequence FMPML leads to improved proton decoupling, while the right-handed modulation sequence FMPMR is ineffective, proves that the efficient decoupling is caused by a secondary resonance effect. PMID- 9211620 TI - Solid-state NMR characterization of copolymers of nylon 11 and nylon 12. AB - Solid-state 13C and 15N NMR spectroscopy, in conjunction with differential scanning calorimetry, wide-angle X-ray diffraction and infrared spectroscopy, were used to characterize a series of nylon 11 and 12 copolymers with mole percentages of nylon 12 monomer of 0, 15, 35, 50, 65, 85, and 100%. Monotonic melting point (Tm) and heat of fusion depressions were observed for the copolymer series with the 65 mol% nylon 12 copolymer having the lowest apparent crystallinity and Tm at 148 degrees C. Solid-state 15N NMR spectra showed a smooth shift of the main peak position for the as-prepared copolymers from 84 ppm for the alpha-form of pure nylon 11 to 89 ppm for the gamma-form of pure nylon 12. Similar behavior was seen for FTIR amide V and VI modes which are also sensitive to the alpha- and gamma-crystal forms. 13C NMR T1 measurements showed that the overall most mobile sample was the 65:35 copolymer. The amide group of the 1:1 copolymer was labelled using 15N-labelled amino acids available through the Gabriel synthesis; an annealed, solution-cast film of this sample showed a T1N value of 349 s, similar to values seen for annealed nylon 11 and nylon 12 homopolymers. The WAXS pattern for the 65 mol% nylon 12 sample showed a sharp peak at 2 theta = 21.3, overlapping a broad peak centered at 2 theta = 21.0. These are consistent with the values seen for gamma-form nylon 12. The 1:1 copolymer (15N labelled) was shown to be polymorphic, like the homopolymers after specific treatments, with a gamma-like phase formed upon solvent casting, and an alpha-like phase dominating for as-polymerized material and precipitated flakes. PMID- 9211621 TI - 1H and 19F magnetic resonance imaging of solid paramagnetic compounds using large magnetic field gradients and Hahn echoes. AB - 1H and 19F spatially resolved echo trains of a number of paramagnetic solids with apparent magnetic moments as high as 10 Bohr magnetons have been successfully obtained in the 58 T m-1 fringe-field gradient of a 9.4 T superconducting magnet by the use of Hahn echoes. Complexes studied include hydrates of Ni(II), Fe(III), Mn(II), and the anhydrous fluorides of Ti(III), Co(II) and Mn(II). Tris 6,6-7,7 8,8,8 heptafluoro-2,2-dimethyl-3,5octane dionato complexes of several lanthanides, Eu, Dy, Ho and Yb, have also been imaged. Additionally, 1H relaxation time measurements have been obtained at 28 MHz with conventional techniques, and several additional T2 values have been obtained in the stray field for both 1H and 19F. PMID- 9211622 TI - A convenient method for calibration of the pulse-length in high field-gradients using Hahn echo-trains. AB - A new method is presented for calibration of the pulse-length in the case that the sample, which can be either a liquid or a solid, is placed in a high field gradient such as it experiences in the stray-, fringe-field of a solenoid. The method employs a pulse-train, with a constant phase, of the form alpha X-tau (alpha X-tau-echo-tau-)n. This produces Hahn echoes which have variable phase in the form of phase-alternations along the echo-train. For alpha = 90 degrees the relative phase-sequence in degrees is (0, 0, 180, 180), and we may characterise this by m = 2, since there are two signals of one phase in each group. For other cases with these phase angles (0 and 180) but different numbers, m, of signals with opposite phase, we have alpha = 180/m, where m is an integer. PMID- 9211623 TI - A solid-state NMR study of the molecular sieve VPI-5 synthesized in the presence of a CTABr surfactant. AB - Silicon-free and silicon-rich large-pore aluminophosphate VPI-5, synthesized with various contents of AIPO-H3 impurity, was studied by Bloch decay (BD) and cross polarization (CP) NMR under magic angle spinning (MAS). The 1H-->31P CP peaks were considerably stronger from AIPO-H3 than from VPI-5. A detailed examination of the CP kinetics and a careful comparison of the CP and BD spectra are prerequisites for the unequivocal interpretation of 1H-->31P CP in porous aluminophosphates. PMID- 9211624 TI - Solid-state NMR studies of interstitial phosphorus atoms in rhodium carbonyl clusters. AB - Pure samples and as-prepared mixtures of Rh9 and Rh10 carbonyl clusters with interstitial P atoms have been studied quantitatively by 31P MAS and 1H-31P CP/MAS NMR. Information on the 31P chemical shift tensor of the Rh9 and Rh10 clusters has been derived from spinning sideband simulations. The chemical shift anisotropy is slightly larger in the Rh10 clusters (340-400 ppm) than in the Rh9 clusters (230-300 ppm), while the asymmetry parameters are similar (eta = 0.1 0.4). The results contribute to the understanding of the relationship between the shielding anisotropy and the structure of the cluster cavity. PMID- 9211625 TI - Enoxaparin, a low-molecular-weight heparin suppresses prothrombin activation more effectively than unfractionated heparin in patients treated for venous thromboembolism. AB - Although unfractionated heparin (UFH) is standard therapy for venous thromboembolism, there is a clinical failure rate of up to 10%. Newer treatment strategies include low-molecular-weight heparins (LMWH). As part of an international study comparing the efficacy and safety of enoxaparin, a LMWH versus UFH in patients with venous thromboembolism, we studied the effects of enoxaparin and UFH on the plasma concentrations of two activation peptides, fragment 1 + 2 (F1 + 2), and thrombin-antithrombin III (TAT) complexes. We hypothesized that enoxaparin would be more effective in suppressing activation of coagulation. Intravenous heparin was given by bolus injection followed by infusion. There were 2 enoxaparin treatment groups (1 mg/kg s.c., bid and 1.5 mg/kg s.c. daily). Plasma samples were obtained from 11 patients in the enoxaparin group and 6 patients in the heparin group prior to and at 6 hour intervals after initiating therapy. Clinical characteristics of the enoxaparin and UFH patient groups were similar. TAT concentrations were not statistically different between groups at any treatment interval. However, plasma F1 + 2 concentrations differed significantly (p < 0.05); concentrations in the enoxaparin group were consistently lower over time than in the UFH group. These results suggest that this LMWH is more effective in suppressing ongoing thrombosis in vivo than UFH in patients with venous thrombosis. PMID- 9211626 TI - Topical application of low molecular weight heparin in a rabbit traumatic anastomosis model. AB - Low molecular weight heparins (LMWHs) are antithrombotic drugs composed of lower molecular weight components of heparin with an apparent molecular weight in the range of 4.0-8.0 KDA. These agents have been used clinically for several years. They have different mechanisms of action compared to heparin, a longer half-life and much higher bioavailability. Anticoagulant drugs such as heparin have been used topically in our previous studies to avoid bleeding complications observed with systemic administration. In this study, low molecular weight heparin (Certoparin, Sandoz) was topically administered in a rabbit ear arterial crush avulsion thrombosis model and compared with heparin. The animals were divided into three groups: LMWH, heparin and saline control groups. In the LMWH group, the patency rate was 71% (10 of 14) at both 1 and 7 days. The patency rate in the heparin group was 95% (19 of 20) at 24 hrs and 80% (16 of 20) at 7 days. In the saline control group, the vessel patency rate was 17% at 24 hrs and 13% at 7 days. Clotting times such as ACT, PT and APTT performed on samples drawn one hour after drug administration were within the normal ranges for both the control and the treatment groups. The results suggest that topical administration of LMWH prevents the occurrence of thrombosis at the traumatic anastomosis site to a similar degree as heparin. PMID- 9211627 TI - Effect of isradipine on in-vivo platelet function. AB - In animal studies calcium channel blockers (CCB's) and especially isradipine, a second generation dihydropyridine, interrupt the sequence of events culminating in the formation of atherosclerotic lesions. The effect of 4 weeks isradipine treatment (5mg daily) on blood pressure and in-vivo platelet function (measured with 111Indium-oxine labeled autologous platelets) were investigated in a randomized, double-blind and placebo controlled trial in 40 patients with mild to moderate hypertension and scintigraphically diagnosed active atherosclerotic lesions of the carotid arteries. The average supine systolic/diastolic blood pressure was significantly reduced at the end of the treatment period in the isradipine group (group 1; p < 0.0001) but remained unchanged in the placebo group (group P). The heart rate was not significantly altered in either group. There were no serious side effects. The platelet uptake ratio (PUR) measured over the atherosclerotic region of the carotid artery on 4 consecutive days before and after treatment decreased significantly in group I from 1.20 to 1.15 (within groups: p < 0.0001) but remained unchanged in group P. Platelet survival increased significantly in group I (mean 5.70 hours, lower quartile 4.50, upper quartile 4.50 hours, within groups: p < 0.0001) and remained unchanged in group P. Isradipine has a beneficial effect on in-vivo platelet function as evidenced by a decreased platelet deposition on vascular lesion sites and an associated prolonged platelet survival in patients with hypertension and active atherosclerotic lesions. PMID- 9211628 TI - Monitoring of recombinant hirudin: assessment of a plasma-based ecarin clotting time assay. AB - Assay conditions of a plasma-based ecarin clotting time (ECT) were evaluated and the precision of the ECT in monitoring plasma levels of r-hirudin assessed. The snake venom enzyme ecarin converts prothrombin to meizothrombin possessing only weak coagulant but strong esterase activity. In r-hirudin containing plasma samples, meizo thrombin is rapidly neutralized by r-hirudin resulting in a dose dependent prolongation of the clotting times. Among the different assay conditions tested, addition of 50 microliters of ecarin (4 U/ml) to 100 microliters of undiluted citrate-anticoagulated plasma gave optimal results with respect to precision and reproducibility. The measuring range of the ECT performed in this way is about 0.02-5.0 micrograms/ml r-hirudin. In vitro studies performed on r-hirudin-spiked plasma samples of 50 healthy individuals demonstrate remarkably low interindividual differences in r-hirudin responsiveness as indicated by CV-values below 5% and 7% at r-hirudin concentrations between 0-3 micrograms/ml and 4-5 micrograms/ml, respectively. The specificity for r-hirudin of the ECT is further demonstrated by the strong correlation (r = 0.94) between the results of a chromogenic assay and the ECT measured r-hirudin concentrations obtained on 67 ex vivo blood samples. Depending on the concentration of r-hirudin the ECT is sensitive to plasma levels of prothrombin. In the absence of r-hirudin the critical prothrombin concentration was found to be 20% but increasing to 60% in the presence of 2.0 micrograms/ml r hirudin. A fibrinogen concentration of 50 mg/dl was found to be the minimal concentration independent of the r-hirudin concentration. The precision of the ECT in measuring plasma levels of r-hirudin is not influenced by treatment with heparin, aprotinin or oral anticoagulants. The data demonstrate that the ECT is a rapid and easily perfor mable clotting assay which allows accurate measurement of r-hirudin plasma levels. ECT-monitored hirudin treatment will help to establish optimal dose regimens that are more efficacious but still as safe as heparin. PMID- 9211629 TI - A simple procedure that increases the specificity of the activated protein C resistance test in samples containing antiphospholipid antibodies. AB - It is well known that plasmas with lupus anticoagulants (LA) may give false low activated protein C (APC) ratios, and these false positive tests are not necessarily corrected by mixing with factor V deficient plasma (FVdef). In the present study, we show that repeating the test after mixing 1 + 1 with pooled normal plasma (NP) instead of mixing with FVdef confirm the presence of the Leiden mutation (FV-Leiden) in patients with antiphospholipid antibodies (APA). Samples from sixteen patients with a low APC-ratio were examined. Eight samples contained APAs, including five samples with LA. RESULTS: The APC-ratios of eleven samples, including four with APAs, became normal when retested after mixing the plasma 1 + 1 with NP. All of them were heterozygous for FV-Leiden. One additional patient, who had partial correction of the APC-ratio, proved to be homozygous for the Leiden mutation. The remaining four samples, all of which had high positive tests for APAs, remained unchanged. One of these four was heterozygous for FV Leiden, while the other three had normal FV. CONCLUSION: Normalisation of a low APC-ratio after dilution 1 + 1 with NP confirms the diagnosis of FV-Leiden. PCR analysis could be reserved for cases not affected by this procedure. PMID- 9211630 TI - Cathepsin A-like activity in thrombin-activated human platelets. Substrate specificity, pH dependence, and inhibitory profile. AB - Cathepsin A-like enzyme released from human platelets by thrombin hydrolyzed at the highest rate Cbz-Phe-Ala, Cbz-Phe-Met and Cbz-Phe-Leu, did not require activators and was inhibited by DFP, DCI and mercurial compounds (mersalyl acid, PCMS, PCMB and HgCl2). The optimum activity of secreted enzyme was at pH 5.0-6.0. Cbz-Glu-Tyr was also hydrolyzed at lower pH with optimum at pH 3.5. These enzymatic properties are the same as those of cathepsin A solubilized from whole platelets and purified from other mammalian cells and tissues. High specific activity of secreted cathepsin A, and a broad pH range of activity may have a significance in extracellular proteolysis in local sites of ischemia. Large portion of cathepsin A-like activity was not secretable by high concentration of thrombin and was sedimented with platelet aggregates. No activity of lysosomal carboxypeptidases B and prolylcarboxypeptidase was detectable in supernatants and pellets of thrombin-stimulated platelets. PMID- 9211631 TI - Mean platelet volume, platelet count and platelet dimensional width during hemodialysis. PMID- 9211632 TI - The prevalence of two genetic traits related to venous thrombosis in whites and African-Americans. PMID- 9211633 TI - Glycocalicin derived peptides inhibit von Willebrand factor binding to platelets. PMID- 9211634 TI - Factor VIIA determination compared to D-Dimer in diagnosis of deep venous thrombosis. PMID- 9211636 TI - Absence of the factor V Leiden mutation in Ethiopians. PMID- 9211635 TI - Platelets inhibit the activity of platelet-activating factor acetylhydrolase in monocyte-derived macrophages. AB - Blood platelets are capable of interacting with monocytes and macrophages and of enhancing various functions of these cells, which are believed to play a role in thrombosis and inflammation. An increase in the uptake of oxidised low density lipoprotein (LDL), in the synthesis of procoagulant tissue factor, thrombospondin and leukotrienes, as well as stimulation of oxygen radical production by platelets has been described (1-5). In circulating blood, a substantial proportion of monocytes was found to be associated with platelets, but the pathophysiological significance of such platelet-monocyte conjugates is not yet clear (6,7). Immigration of monocytes into the arterial intima and their differentiation into macrophages are initial steps in the development of an atherosclerotic lesion (8). During differentiation, there is a tremendous increase in the activity and secretion of the enzyme PAF acetylhydrolase (PAF = platelet-activating factor = 1-O-alkyl-2-acetyl-sn-glycero-3-phosphocholine) (9,10), and there is some evidence that this enzyme may contribute to the development of atherosclerosis. It cleaves PAF, and the remaining lyso-PAF is chemotactic for monocytes (11). Furthermore it also acts on oxidised low density lipoproteins and enhances their uptake into macrophages (12,13). We were therefore interested in investigating whether platelets may modulate the differentiation of monocytes into macrophages and the activity of PAF acetylhydrolase. PMID- 9211637 TI - How viruses hide from T cells. PMID- 9211638 TI - Host resistance to malaria runs into swampy water. PMID- 9211639 TI - Quantification of the reservoir of HIV-1. PMID- 9211640 TI - Adhesion and invasion by the pathogenic neisseria. PMID- 9211641 TI - Miles of isles. PMID- 9211642 TI - The role of Actinobacillus actinomycetemcomitans in the pathogenesis of periodontal disease. AB - Periodontal disease consists of a constellation of complex bacterium-host cell interactions. One example of these oral pathogens, Actinobacillus actinomycetemcomitans, has an arsenal of putative virulence determinants that account for its potent periodontopathogenicity. Of these determinants, invasion of host cells and leukocytotoxicity have been studied extensively. PMID- 9211643 TI - Recent structural solutions for antibody neutralization of viruses. AB - Structural studies of virus-antibody interactions can offer insights into mechanisms of virus neutralization and immune escape. Crystallography and cryoelectron microscopy can reveal the structural relationship between receptor binding sites and neutralization epitopes, as well as whether molecular rearrangements occur upon antibody binding. PMID- 9211644 TI - Bacterial resistance to aminoglycoside antibiotics. AB - The aminoglycoside antibiotics are broad-spectrum antibacterial compounds that are used extensively for the treatment of many bacterial infections. In view of the current concerns over the global rise in antibiotic-resistant microorganisms, there has been renewed interest in the mechanisms of resistance to the aminoglycosides, including the superfamily of aminoglycoside-modifying enzymes. PMID- 9211645 TI - Beyond vancomycin: new therapies to meet the challenge of glycopeptide resistance. AB - The incidence of infections caused by resistant Gram-positive pathogens is increasing, while emergence of vancomycin resistance is reducing the number of therapeutic options. New agents are being rapidly evaluated as candidates to replace vancomycin; some of the most promising include semisynthetic glycopeptides, quinupristin-dalfopristin, oxazolidinones and everninomycins. Alternative strategies, including immunization and therapeutic vaccines, may also have a role. PMID- 9211646 TI - Feline heartworm (Dirofilaria immitis) infection: detection of specific IgG for the diagnosis of occult infections. AB - Sera from 54 cats, 53 asymptomatic and one symptomatic (chronic dyspnoea and coughing), living in a hyper-endemic area for canine heartworm (Dirofilaria immitis) infection were studied to evaluate the reliability of two ELISA-based antibody tests coupled with both somatic (ELISASA) and excretory/secretory (ELISAE/S) antigens. All cats were examined by echocardiography and radiography. In addition, an ELISA-based test to detect adult female heartworm circulating antigens and a modified Knott test for microfilariae in the blood were carried out on all cats. No cat was positive for microfilariae in the blood. Heartworms were visualized in 12 of 54 cats by echocardiography. Of these, three asymptomatic cats and the symptomatic one had radiographic signs of infection and were the only ones positive for heartworm circulating antigens. All sera except two were positive when analyzed in ELISA(SA). In ELISA(E/S), nine sera were positive but three were negative. No sera from the 42 echocardiography-negative cats was positive in ELISA(E/S), but 11 were positive in ELISA(SA). Western blot analyses with somatic antigens of sera from echocardiography-positive cats showed at least four bands of recognition between 19 and 30 kDa and one of about 40 kDa. With E/S antigen, a large band of about 22 kDa and one of about 25 kDa were recognized; these appear to be most specific. PMID- 9211647 TI - Comparison of the persistent activity of ivermectin, abamectin, doramectin and moxidectin in cattle in Zambia. AB - The persistent efficacy of four commercially available macrocyclic lactones (ML) in maintaining reduced faecal egg counts in cattle grazing naturally infested pastures was evaluated in 44 zebu animals aged 1-2 years in Zambia. The study started in February (rainy season) when the strongyle egg output was increasing. Four days before the start of the trial, all animals were treated with a double dose of oxfendazole. They were then divided into five groups which were again treated on day 0. Groups A, D, I and M received 0.2 mg kg-1 of abamectin, doramectin, ivermectin and moxidectin, respectively. Animals of group C received albendazole (7.5 mg kg-1). Faecal samples were collected twice a week for egg counts and larval differentiation. Faecal egg counts in the C group increased from day 21 onwards and plateaued from day 42 between 180 and 380 eggs per gram. The main genera found in cultures were Cooperia (90%) and Haemonchus (7%). Faecal egg excretion in groups M, A, D and I started on day 35, 42, 42 and 45, respectively. Subsequently and until day 84, average counts in these four groups were always significantly lower than in group C. Compared with albendazole, all four ML gave over 95% reduction in cumulative faecal egg counts for 42 days after treatment. The percentage efficacy was still over 84% by day 84 when an average cumulative egg count of 11320 eggs per gram faeces was calculated in group C. In addition, there was no significant difference in efficacy between the four ML groups at any of the sampling dates. During the trial no significant difference in weight gain between any of the groups was observed. PMID- 9211648 TI - Transmission of some species of internal parasites in horses born in 1993, 1994, and 1995 on the same pasture on a farm in central Kentucky. AB - Data are presented on the last 3 years of a 7-year study (1989-1995) on transmission of natural infections of internal parasites in horse foals (n = 27) born in 1993, 1994, and 1995 on the same pasture on a farm in central Kentucky. The foals were in a closed breeding herd of horses. Research on the first 4 years (1989-1992) of the study was published earlier (Lyons et al., 1991, 1994). Thirty five species of endoparasites were identified, including 24 species of small strongyles. Monthly, seasonal, and host-age transmission patterns were elucidated for the parasites. Comparison of data between the first 4 years and last 3 years of the study indicates similarities, but also differences, including an increase in prevalence and numbers of Thelazia lacrymalis and Anoplocephala perfoliata. PMID- 9211649 TI - The repeatability of faecal egg counts in Polish Wrzosowka sheep. AB - The number of nematode eggs in the faeces was estimated in Polish Wrzosowka sheep, in the spring and autumn of 1993 and again in 1994. The sheep had been naturally infected. The dominant species were Haemonchus contortus and Teladorsagia circumcincta, but Trichostrongylus spp., Cooperia curticei, Nematodirus spp. and Chabertia ovina were also present. Anthelmintics were not used. Egg counts were skewed, with a range of 0-4100 eggs g-1 (EPG); most sheep had egg counts below 100 EPG. Egg counts were approximately four times higher in spring than in autumn. Repeatability values within a season were all significant and positive. The repeatability of egg counts between seasons was estimated from the correlation between the mean transformed value in spring and in autumn, and was 0.52 in 1993 (P < 0.001) and 0.41 in 1994 (P < 0.05). The results show that animals with higher than average values in spring are likely to have higher than average values in autumn, and suggest that similar mechanisms regulated egg counts in both seasons even though egg counts were much lower in autumn. PMID- 9211650 TI - Infestation of sheep dung by nematophagous fungi and implications for the control of free-living stages of gastro-intestinal nematodes. AB - A field trial was conducted to assess the rate at which dung becomes infested by fungi which parasitise nematodes (nematophagous fungi) after deposition. Sheep dung was placed on field plots of bare ground, ryegrass (Lolium perenne), browntop (Agrostis capillaris) and white clover (Trifolium repens) in summer (February) and autumn (April), and subsamples were examined at intervals for the presence of nematophagous fungi. Nematophagous fungi occurred in 71% of 129 samples recovered in February and 57% of 58 samples recovered in April. Arthrobotrys oligospora, Monacrosporium candidum and Nematoctonus spp. were the most frequently isolated nematode-trapping fungi in both seasons. The endoparasitic nematophagous fungus Harposporium leptospira also occurred frequently in dung deposited in February, but not April. Fungi entered dung quickly, with 83% and 58% of dung samples containing nematophagous fungi at 3 days after deposition in February and April, respectively. The percentage of dung infested by nematophagous fungi on plots of bare ground, ryegrass, white clover and browntop was 76%, 75%, 61% and 55%, respectively. Results suggest that a number of species of nematophagous fungi are able to enter dung soon after deposition on a variety of types of ground cover. PMID- 9211651 TI - Epidemiology of nematodosis in Romney lambs selectively bred for resistance or susceptibility to nematode infection. AB - Field trials were undertaken to compare nematode population dynamics, lamb productivity and levels of breech soiling in experimental flocks of Romney lambs selectively bred for increased resistance or susceptibility to nematode infection. In each year of the 2 year study, spring-born ewe lambs derived from Wallaceville Animal Research Centre's divergent nematode-resistant and nematode susceptible breeding lines were grazed as separate flocks on matched farmlets from weaning (at 3 months old) until they were approximately 10-11 months old. Allocation of farmlets was reversed between Years 1 and 2 of the study to account for any possible paddock-related effects. Within each year both flocks were subjected to identical management conditions, including anthelmintic treatment (which was administered only when the overall mean faecal worm egg count measured across both genotypes reached 1500 eggs g-1). In both years, by mid-autumn (April) nematode larval infestation levels on pasture were approximately 5-6-fold greater on the farmlet grazed by susceptible (S) genotype lambs than on that grazed by their resistant (R) counterparts (Year 1: 2506 cf. 544 larvae kg-1 herbage; Year 2: 431 cf. 74 larvae kg-1 herbage). This led to 51-fold and 56-fold differences in faecal egg count between R and S lambs by late autumn (May) and winter (July) in Years 1 and 2, respectively. Although mean growth rates were similar in the R and S lambs over summer (while pasture infestation levels on the farmlets were still in the process of diverging), significantly higher growth rates occurred in the R than in the S lambs over autumn-winter in both years of the study (P < 0.01). In contrast, no significant differences in growth rate occurred in either year between male lambs derived from the nematode-resistant and nematode-susceptible breeding lines which were grazed together on another area of the Wallaceville farm from weaning until late autumn. Despite the substantially lower pasture infestation levels encountered by the R ewe lambs, they nevertheless temporarily suffered more breech soiling (dags) than their S counterparts (P < 0.01) in both years. Yearling fleece-weights of the R and S genotypes did not differ significantly in either year. Although the results of our study confirmed that there are potentially significant epidemiological benefits to be derived from breeding sheep for resistance to nematode infection, these benefits did not appear to be associated with large advantages in animal performance. Further work is needed to establish how these results should be interpreted with respect to anthelmintic drench requirements of genetically resistant animals. PMID- 9211652 TI - Ultrastructure of Anaplasma marginale with an inclusion appendage, isolated in Minas Gerais State, Brazil. AB - This is the first report on the occurrence and isolation of a strain of Anaplasma marginale with an inclusion appendage in Brazil. The inclusion appendage presented longitudinal electron-dense striations and did not originate directly from the body of the rickettsia but from an electron-dense complex located at the junction of the inclusion membrane and inclusion appendage. The inclusion appendage remained in the host cell after the Anaplasma inclusion appeared to be leaving the red blood cell. Other ultrastructures of this rickettsia are described and its epidemiological importance is discussed. PMID- 9211653 TI - Hepatozoon canis infection of wild carnivores in Brazil. AB - Hepatozoon canis was diagnosed in a crab-eating fox (Cerdocyon thous) found on a highway in the region of Botucatu, Sao Paulo, Brazil, after being hit by a car. The fox had bilateral fractures of the olecranon, which was corrected by osteosynthesis. Hematologic findings included a neutrophilia, eosinophilia, monocytosis and mild anemia. In the Leishman-stained blood film, gametocytes of Hepatozoon canis in neutrophils were identified measuring 9.1 +/- 0.54 x 5.3 +/- 0.46 microns. PMID- 9211654 TI - Comments on the paper 'World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) Second Edition of Guidelines for Evaluating the Efficacy of Anthelmintics in Ruminants (Bovine, Ovine, Caprine)'. PMID- 9211655 TI - Metabolism of the calcium antagonist, mibefradil (POSICOR, Ro 40-5967). Part II. Metabolism in hepatic microsomes from rat, marmoset, cynomolgus monkey, rabbit and man. AB - 1. The calcium antagonist, mibefradil, is converted to some 30 metabolites after incubation with hepatic microsomes from the rat, marmoset, cynomolgus monkey, rabbit and man. 2. The wide inter-species differences in metabolic profile stem mainly from variations in the activity of the microsomal esterase, which hydrolyses the ester side-chain of mibefradil to give the alcohol metabolite, Ro 40-5966. Hydrolysis is especially marked in the cynomolgus monkey and rabbit, less in man and least in the rat and marmoset. 3. The biotransformation of this alcohol metabolite by cytochromes P450 is more facile than that of the parent compound, leads to fewer metabolites and the metabolic profiles in all species are similar. 4. The most important cytochrome P450-mediated metabolic process in microsomes in all species is hydroxylation at the benzylic carbon atom of the tetrahydronaphthyl group; further oxidation of the resultant secondary alcohol to a ketone also occurs. These reactions indicate the route of the biosynthetic pathway which leads to the major, naphthyl-glucuronide metabolites previously isolated from rat bile. 5. Dealkylation of the tertiary amino group is also important and leads to compounds lacking either the N-methyl group or the propylbenzimidazole moiety. 6. Hydroxylation of the benzimidazole ring at both the 4- and 5-positions is largely restricted to mibefradil and does not occur to a significant extent with Ro 40-5966. PMID- 9211656 TI - Metabolism of the calcium antagonist, mibefradil (POSICOR, Ro 40-5967). Part III. Comparative pharmacokinetics of mibefradil and its major metabolites in rat, marmoset, cynomolgus monkey and man. AB - 1. The metabolism of mibefradil has been examined in rat, marmoset, cynomolgus monkey and man after single and multiple oral administration. 2. Metabolites typically represent between 50 and 80% of the circulating drug-related material after single oral doses of mibefradil to man, rat and marmoset. They arise by a combination of enzymatic processes: cytochrome P450-mediated oxidation at saturated and unsaturated carbon atoms, cytochrome P450-catalysed dealkylation and hydrolysis of the ester side-chain. 3. Plasma levels of mibefradil in the cynomolgus monkey are extremely low as a result of very efficient first-pass hydrolysis of its side-chain to give the corresponding alcohol. Steady-state concentrations of this metabolite are comparable with those of the parent drug in man and marmoset, but are relatively low in rat plasma. 4. Hydroxylation at the benzylic carbon of the tetrahydronaphthyl ring leads to further important metabolites in primates, whereas the products of O- and N-demethylation are found in small amounts in all four species. 5. Estimates of the exposure of the various species to the principal metabolites indicate that the choice of the rat, marmoset and cynomolgus monkey for the toxicological assessment of mibefradil was appropriate. PMID- 9211657 TI - Purification and characterization of hepatic glutathione transferases from an insectivorous marsupial, the brown antechinus (Antechinus stuartii). AB - 1. Five unique glutathione transferase isoenzymes were purified from the hepatic cytosol of an insectivorous marsupial, the brown antechinus. The purified GSTs were characterized by structural and catalytic properties including apparent molecular weight and isoelectric point, specificity towards model substrates, kinetic parameters, sensitivity to inhibitors and cross-reactivity with antisera raised against human GSTs. 2. An alpha class GST, Antechinus GST 1-1, predominated in the hepatic cytosol, representing 71% of the total GST purified. The substrate specificity of Antechinus GST 1-1 was similar to that of other alpha class GSTs, particularly with respect to its high activity with cumene hydroperoxide. The mu class was represented by three GST isoenzymes, Antechinus GST 3-3, GST 3-4 and GST 4-4. These isoenzymes represented 8, 2 and 10% of the total GST purified respectively. A single GST, Antechinus GST 22, belonged to the pi class of GSTs and represented 12% of the total GST purified. The hepatic GST isoenzyme ratio (by class) observed in the brown antechinus was more similar to that observed in the human than in rat. 3. A previous study investigating a herbivorous marsupial, the brushtail possum (Trichosurus vulpecula) also identified a predominant hepatic GST belonging to the alpha class and displaying peroxidase activity. The evolutionary conservation of a similar predominant GST isoenzyme in these marsupials suggests that they play an important role in the detoxication metabolism of these unique mammals. PMID- 9211658 TI - The isoflavones equol and genistein do not induce xenobiotic-metabolizing enzymes in mouse and in human cells. AB - 1. In view of the anticarcinogenic effects of the isoflavonoid genistein and the known ability of various flavonoids to induce carcinogen-metabolizing enzymes, the ability of the isoflavonoids genistein and equol to induce these enzymes was studied in mouse. 2. In comparison with induction by the positive control beta naphthoflavone (40 mg/kg i.p. 4 days) neither genistein or equol (0.4-40 mg/kg i.p. 4 days) gave a significant induction of ethoxyresorufin O-deethylase, p nitrophenol oxidase or immunoreactive CYP1A2 or CYP2E1. There was also no induction of erythromycin-N-demethylase or elevation of immunoreactive CYP3A1. 3. In contrast with the induction by beta-naphthoflavone of glutathione S transferase protein and activity towards 1-chloro-2,4-dinitrobenzene, neither isoflavone exhibited such induction. 4. Response elements for human CYP1A1, glutathione S-transferase lambda a and the xenobiotic response element (XRE) in HepG2 cells were activated by beta-naphthoflavone but not by genistein or equol. 5. The isoflavones have been found not to induce a range of enzymes involved in carcinogen metabolism. The lack of enzyme induction differs from the characteristics of many other flavonoids. The results do not support the monofunctional induction of GST as a basis of anticarcinogenicity. PMID- 9211659 TI - Absorption, metabolism and excretion after oral administration of a new Ca antagonist, 14C-benidipine hydrochloride to man. AB - 1. In healthy male volunteers, the absorption, metabolite profiles and excretion of 14C-benidipine hydrochloride, a new Ca antagonist, were investigated after oral administration at a dose of 8 mg. 2. 14C-benidipine hydrochloride was rapidly absorbed, and the plasma concentration of radioactivity and unchanged drug reached a maximum of 71.2 ng eq./ml at 1.1 h and 2.56 ng/ml at 0.6 h respectively, and then declined bi-exponentially. The half-life in the elimination phase was 14.7 and 5.3 h respectively, AUC of unchanged drug was low, about 1% of that of radioactivity. 3. Five days after administration, 36.4% of the administered radioactivity was excreted in urine and 58.9% in faeces. 4. The metabolite profiles in plasma, urine and faeces were analysed by hplc. At 1 h after administration the predominant metabolites in plasma were M9 and M2, which accounted for 13.8 and 8.2% of the radioactivity respectively, whereas unchanged drug represented 1.2%. Predominant metabolites in urine 12 h after administration were M3 and M8, which accounted for 2.22 and 2.21% of the administered radioactivity respectively. Metabolites excreted in faeces 120 h after administration were very complex and poorly separated by hplc and could not be characterized: unchanged drug was not detected in the faeces. PMID- 9211660 TI - Pharmacokinetics of 2-naphthol following intrapericardial administration, and formation of 2-naphthyl-beta-D-glucoside and 2-naphthyl sulphate in the American lobster, Homarus americanus. AB - 1. Following a 0.25-mg/kg intrapericardial dose of the phenolic compound, 2 naphthol, to the American lobster, Homarus americanus, a two-compartment model best described the disposition of parent [14C]-2-naphthol in the haemolymph. Male and female lobsters had similar alpha-phase half lives of 26 +/- 19 min (mean +/- SD, n = 4) and 29 +/- 15 min respectively. The beta-phase half lives were significantly longer in males, 63.9 +/- 30.9 h, than in females, 30.6 +/- 6.8 h (p < 0.05). The total body clearance for females was 26.4 +/- 6.5 ml x h-1 x kg-1 and was higher than that of males, 11.1 +/- 5.9 ml x h-1 x kg-1 (p < 0.05). 2. 2 Naphthol was converted to 2-naphthyl-beta-D-glucoside (major metabolite) and 2 naphthyl sulphate (minor metabolite) such that at 24 h 39-60.6% of the radioactivity in haemolymph was 2-naphthyl-beta-D-glucoside, 38.6-58.9% 2 naphthol and 0.5-4% 2-naphthyl sulphate. 3. The 2-naphthol-derived radioactivity was > 99% bound to haemolymph proteins at 1 min and > 90% bound at 1 day after the dose, indicating that both 2-naphthol and 2-naphthyl-beta-D-glucoside were highly protein bound. 4. 2-Naphthyl-beta-D-glucoside was slowly eliminated from haemolymph in both males and females, with elimination half lives of 34-78 h. 2 Naphthyl-beta-D-glucoside was the major metabolite in urine samples collected at 5 days after the dose. Hepatopancreas and antennal gland contained glucosidase activities, and the long half life of 2-naphthyl-beta-D-glucoside could be explained by conjugation deconjugation cycling. 5. 2-Naphthyl sulphate was eliminated from haemolymph with a half-life < 10 h and was excreted in urine. PMID- 9211661 TI - Intraperitoneal and intraportal administration of droloxifene to the Sprague Dawley rat: assessing the first-pass effect. AB - 1. Employing droloxifene as a probe substrate, we have compared the use of intraperitoneal injection and intraportal infusion, where the rate and duration of intraportal drug administration were designed to approximate those observed after oral drug delivery, as methods of discriminating between high first-pass hepatic extraction and poor oral absorption. 2. Intraperitoneal injection of droloxifene (1 mg/kg) yielded an AUC0-omega approximately twice that observed following intraportal infusion or oral delivery of equal doses. 3. Our findings suggest that hepatic first-pass metabolism may have been saturated following intraperitoneal drug administration due to the rapid rate of absorption and the corresponding high drug concentrations achieved. 4. Application of a model in which intraportal drug infusion rates are designed to mimic the oral absorption rate appears warranted under such circumstances. PMID- 9211662 TI - [Systematic therapy of vaginal mycoses. Three quarters of women are affected]. PMID- 9211664 TI - [Crataegus in cardiac insufficiency--taking a current position]. PMID- 9211663 TI - [Hearing loss--current data on diagnosis and therapy]. PMID- 9211665 TI - [Immunoactive peptides in chronic polyarthritis]. PMID- 9211666 TI - [Advance through research]. PMID- 9211667 TI - [New proton pump inhibitors. Advances in the therapy of acid-induced diseases]. PMID- 9211668 TI - [Depression. New perspectives in diagnosis and therapy. Erlangen, 21 November 1992]. PMID- 9211669 TI - [Gabapentin--a new anti-epileptic agent]. PMID- 9211670 TI - [Current therapeutic possibilities with tricyclic antidepressive agents]. PMID- 9211671 TI - [Herbal remedies against depression]. PMID- 9211672 TI - [Ambulatory surgery--pre- and postoperative care]. PMID- 9211673 TI - [Osteoporosis: calcium and vitamin D3 are the basis for prevention and therapy. A national disease that should not be]. PMID- 9211674 TI - [BCG therapy of superficial bladder cancer. Current status of clinical aspects and research]. PMID- 9211675 TI - Trial of calcium to prevent preeclampsia. AB - BACKGROUND: Previous trials have suggested that calcium supplementation during pregnancy may reduce the risk of preeclampsia. However, differences in study design and a low dietary calcium intake in the populations studied limit acceptance of the data. METHODS: We randomly assigned 4589 healthy nulliparous women who were 13 to 21 weeks pregnant to receive daily treatment with either 2 g of elemental calcium or placebo for the remainder of their pregnancies. Surveillance for preeclampsia was conducted by personnel unaware of treatment group assignments, using standardized measurements of blood pressure and urinary protein excretion at uniformly scheduled prenatal visits, protocols for monitoring these measurements during the hospitalization for delivery, and reviews of medical records of unscheduled outpatient visits and all hospitalizations. RESULTS: Calcium supplementation did not significantly reduce the incidence or severity of preeclampsia or delay its onset. Preeclampsia occurred in 158 of the 2295 women in the calcium group (6.9 percent) and 168 of the 2294 women in the placebo group (7.3 percent) (relative risk, 0.94; 95 percent confidence interval, 0.76 to 1.16). There were no significant differences between the two groups in the prevalence of pregnancy-associated hypertension without preeclampsia (15.3 percent vs. 17.3 percent) or of all hypertensive disorders (22.2 percent vs. 24.6 percent). The mean systolic and diastolic blood pressures during pregnancy were similar in both groups. Calcium did not reduce the numbers of preterm deliveries, small-for-gestational-age births, or fetal and neonatal deaths; nor did it increase urolithiasis during pregnancy. CONCLUSIONS: Calcium supplementation during pregnancy did not prevent preeclampsia, pregnancy associated hypertension, or adverse perinatal outcomes in healthy nulliparous women. PMID- 9211676 TI - Vitamin D-receptor gene polymorphisms and bone density in prepubertal American girls of Mexican descent. AB - BACKGROUND: Bone mass is under strong genetic control, and recent studies in adults have suggested that allelic differences in the gene for the vitamin D receptor may account for inherited variability in bone mass. We studied the relations of the vitamin D-receptor genotype to skeletal development and variation in the size, volume, and density of bone in children. METHODS: We identified three allelic variants of the vitamin D-receptor gene using the polymerase chain reaction and three restriction enzymes (ApaI, BsmI, and TaqI) in 100 normal prepubertal American girls of Mexican descent. We then determined the relations of the different vitamin D-receptor genotypes (AA, Aa, aa, BB, Bb, bb, TT, Tt, and tt) to the cross-sectional area, cortical area, and cortical bone density of the femoral shaft and the cross-sectional area and density of the lumbar vertebrae. RESULTS: The vitamin D-receptor genotype was associated with femoral and vertebral bone density. Girls with aa and bb genotypes had 2 to 3 percent higher femoral bone density (P=0.008 and P=0.04, respectively) and 8 to 10 percent higher vertebral bone density (P=0.01 and P=0.03, respectively) than girls with AA and BB genotypes. There was no association between the cross sectional area of the vertebrae or the cross-sectional or cortical area of the femur and the vitamin D-receptor genotype. The chronologic age, bone age, height, weight, body-surface area, and body-mass index did not differ significantly among girls with different vitamin D-receptor genotypes. CONCLUSIONS: Vitamin D receptor gene alleles predict the density of femoral and vertebral bone in prepubertal American girls of Mexican descent. PMID- 9211677 TI - Treatment of cytomegalovirus retinitis with a sustained-release ganciclovir implant. The Ganciclovir Implant Study Group. AB - BACKGROUND: Sustained-release, intraocular implants that deliver ganciclovir are an alternative method for the treatment of cytomegalovirus retinitis in patients with the acquired immunodeficiency syndrome (AIDS). METHODS: We conducted a randomized study of 188 patients with AIDS and newly diagnosed cytomegalovirus retinitis. The patients were randomly assigned to treatment with an implant delivering 1 microg of ganciclovir per hour, an implant delivering 2 microg of ganciclovir per hour, or intravenous ganciclovir. The primary outcome we studied was progression of cytomegalovirus retinitis. RESULTS: The median time to progression of retinitis was 221 days with the 1-microg-per-hour implant (75 eyes), 191 days with the 2-microg-per-hour implant (71 eyes), and 71 days with ganciclovir administered intravenously (76 eyes; P<0.001). The risk of progression of retinitis was almost three times as great among patients treated with intravenous ganciclovir as among those treated with a ganciclovir implant (risk ratio, 2.8; P<0.001). However, the risk of disease in the initially uninvolved eye was lower with intravenous ganciclovir than with a ganciclovir implant (risk ratio, 0.5; P=0.19). Patients treated with intravenous ganciclovir were also less likely to have extraocular cytomegalovirus infections (0, vs. 10.3 percent in the two implant groups; P=0.04). CONCLUSIONS: For the treatment of cytomegalovirus retinitis, the sustained-release ganciclovir implant is more effective than intravenous ganciclovir, but patients treated with a ganciclovir implant alone remain at greater risk for the development of cytomegalovirus disease outside of the treated eye. PMID- 9211678 TI - Effect of testosterone and estradiol in a man with aromatase deficiency. PMID- 9211679 TI - Images in clinical medicine. Sand as a foreign body. PMID- 9211680 TI - Autism. PMID- 9211681 TI - Treatment of cytomegalovirus retinitis in patients with the acquired immunodeficiency syndrome. PMID- 9211682 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 21-1997. A 67-year-old woman with a progressive movement disorder and a left-upper-quadrant mass. PMID- 9211683 TI - Prevention or early treatment of preeclampsia. PMID- 9211684 TI - Vitamin D-receptor genotypes and bone density. PMID- 9211685 TI - New method for analysis of pyrethroid insecticides: esfenvalerate, cis permethrin, and trans-permethrin, in surface waters using solid-phase extraction and gas chromatography. PMID- 9211686 TI - Dermal and respiratory exposure of mixers/sprayers to acephate, methamidophos, and endosulfan during tobacco production. PMID- 9211687 TI - Granular terbufos exposure and cleanup of glove materials. PMID- 9211688 TI - Fish contamination and human exposure to mercury in the Tapajos River Basin, Para State, Amazon, Brazil: a screening approach. PMID- 9211689 TI - Occurrence of pesticides in the Arno River and in potable water--a survey of the period 1992-1995. PMID- 9211690 TI - Lead levels in Argentine market wines. PMID- 9211691 TI - Residues of metolachlor herbicide in soil and potato tubers under Indian tropical conditions. PMID- 9211692 TI - Influence of pesticides on methane oxidation in a flooded tropical rice soil. PMID- 9211694 TI - Photodechlorination pathways of non-ortho substituted PCBs by ultraviolet irradiation in alkaline 2-propanol. PMID- 9211693 TI - Comparison of three bioremediation agents for mineralization and transformation of pentachlorophenol in soil. PMID- 9211695 TI - Cardiorespiratory and neuromuscular effects of O-ethyl S--2-(diisopropylamino) ethyl- methylphosphonothioate (VX) in rats. PMID- 9211696 TI - Effect of N-nitrosodiethylamine on lipid peroxidation and antioxidant enzymes in rat liver mitochondria: protective role of antioxidants. PMID- 9211697 TI - White blood cell count as an indicator of formaldehyde exposure. PMID- 9211698 TI - Changes in adenosine triphosphate (ATP) concentration and its activity in murine tissue after thallium administration. PMID- 9211699 TI - Cytotoxicity of methacrylonitrile. PMID- 9211700 TI - Assessment of metal uptake and genetic damage in small mammals inhabiting a fly ash basin. PMID- 9211701 TI - Recovery of brain cholinesterases of brown-headed cowbirds from organophosphorus intoxication: effect of environmental temperature. PMID- 9211702 TI - Ammonia tolerance of the bivalve Mulinia lateralis sublethal sediment toxicity test. PMID- 9211703 TI - Growth response of freshwater algae to continuous flow of terbutryn. PMID- 9211704 TI - Siderophore production in two species of nostoc as influenced by the toxicity of nitrophenolics. PMID- 9211705 TI - Acute toxicity of organophosphorus insecticides to marine invertebrates. PMID- 9211706 TI - Comparative tolerance of three populations of the freshwater shrimp (Paratya australiensis) to the organophosphate pesticide, chlorpyrifos. PMID- 9211707 TI - Pressure and temperature effects on growth and viability of the hyperthermophilic archaeon Thermococcus peptonophilus. AB - We studied the effects of high temperatures and elevated hydrostatic pressures on the physiological behavior and viability of the extremely thermophilic deep-sea archaeon Thermococcus peptonophilus. Maximal growth rates were observed at 30 and 45 MPa although no significant increases in cell yields were detected. Growth at 60 MPa was slower. The optimal growth temperature shifted from 85 degrees C at 30 MPa to 90-95 degrees C at 45 MPa. Cell viability during the stationary phase was also enhanced under high pressure. A trend towards barophily at pressures greater than those encountered in situ at the sea floor was demonstrated at increasing growth temperatures. The viability of cells during starvation, at high temperature (90, 95 degrees C), and at low temperature (10 degrees C) was enhanced at 30 and 45 MPa as compared to atmospheric pressure. These results show that the extremely thermophilic archaeon T. peptonophilus is a barophile. PMID- 9211708 TI - Characterization of the intron-containing citrate synthase gene from the alkanotrophic yeast Candida tropicalis: cloning and expression in Saccharomyces cerevisiae. AB - Citrate synthase, an essential enzyme of the tricarboxylic acid cycle in mitochondria, was purified from acetate-grown Candida tropicalis. Results from SDS-PAGE and gel filtration showed that this enzyme was a dimer composed of 45 kDa subunits. A citrate synthase cDNA fragment was amplified by the 5'-RACE method. Nucleotide sequence analysis of this cDNA fragment revealed that the deduced amino acid sequence contained an extended leader sequence which is suggested to be a mitochondrial targeting signal, as judged from helical wheel analysis. Using this cDNA probe, one genomic citrate synthase clone was isolated from a yeast lambdaEMBL3 library. The nucleotide sequence of the gene encoding C. tropicalis citrate synthase, CtCIT, revealed the presence of a 79-bp intron in the N-terminal region. Sequences essential as yeast splicing motifs were present in this intron. When the CtCIT gene including its intron was introduced into Saccharomyces cerevisiae using the promoter UPR-ICL, citrate synthase activity was highly induced, which strongly indicated that this intron was correctly spliced in S. cerevisiae. PMID- 9211709 TI - Thiorhodococcus minus, gen. nov., sp. nov., A new purple sulfur bacterium isolated from coastal lagoon sediments. AB - A new marine phototrophic purple sulfur bacterium (strain CE2203) was isolated in pure culture from a man-made coastal lagoon located on the Atlantic coast (Arcachon Bay, France). Single cells were coccus-shaped, did not contain gas vesicles, and were highly motile. Intracellular photosynthetic membranes were of the vesicular type. Bacteriochlorophyll a and carotenoids of the normal spirilloxanthin series were present as photosynthetic pigments. Hydrogen sulfide, thiosulfate, elemental sulfur, and molecular hydrogen were used as electron donors during photolithotrophic growth under anoxic conditions, while carbon dioxide was utilized as carbon source. Acetate, propionate, lactate, glycolate, pyruvate, fumarate, succinate, fructose, sucrose, ethanol, and propanol were photoassimilated in the presence of hydrogen sulfide. During growth on sulfide, elemental sulfur globules were stored inside the cells. Chemotrophic growth under microoxic conditions in the dark was possible. The DNA base composition was 66.9 mol% G+C. Comparative sequence analysis of the 16S rRNA gene confirmed the membership of strain CE2203 in the family Chromatiaceae. Morphological characteristics of strain CE2203 indicated a close affiliation to the genera Thiocystis and Thiocapsa. However, the phylogenetic treeing revealed no closer relationship to Thiocystis spp. than to Thiocapsa roseopersicina or other known members of the Chromatiaceae. Consequently, strain CE2203 is proposed as the type strain of a new genus and species, Thiorhodococcus minus gen. nov., sp. nov. PMID- 9211711 TI - Evidence for an isomeric muconolactone isomerase involved in the metabolism of 4 methylmuconolactone by Alcaligenes eutrophus JMP134. AB - An enzyme specifically induced during 4-methylmuconolactone metabolism by Alcaligenes eutrophus JMP 134 and that exhibited muconolactone isomerizing activity was purified to homogeneity. The enzyme, involved in the isomerization of 3-methylmuconolactone had a high degree of sequence similarity with muconolactone isomerase of Alcaligenes eutrophus JMP 134 and other previously described muconolactone isomerases of the 3-oxoadipate pathway. Kinetic analysis showed that the enzyme has a substrate spectrum and a reaction mechanism similar to those of the muconolactone isomerase, but that it has distinct kinetic properties. PMID- 9211710 TI - The characterization of the nv-gvpACNOFGH gene cluster involved in gas vesicle formation in Natronobacterium vacuolatum. AB - The haloalkaliphilic archaeon Natronobacterium vacuolatum forms cylinder-shaped gas vesicles throughout the growth cycle when grown in media containing 15-25% NaCl. Cells cultivated in media containing 13% NaCl are, however, gas-vesicle free. The major gas vesicle structural protein, nv-GvpA, was detected by an antiserum raised against the gas vesicles of Haloferax mediterranei; the antiserum reacted with an 8.3-kDa protein in samples containing cell extracts or purified gas vesicles of N. vacuolatum. The gene encoding nv-GvpA was isolated together with six additional gvp genes; these genes are arranged consecutively in a cluster as nv-gvpACNOFGH and are cotranscribed. Transcript analysis by primer extension revealed only one start site three nucleotides upstream of the nv-gvpA reading frame. This arrangement of gvp genes differs from that of the gas-vesicle encoding genes in Halobacterium salinarium and Hf. mediterranei. The comparison of the deduced Gvp protein sequences indicated similarities with the respective halobacterial Gvp proteins, with GvpA exhibiting the highest degree of conservation (97-100%). The second gas vesicle structural protein, nv-GvpC, was 150-250 amino acids longer than all other halobacterial GvpC proteins and was much less conserved (48-73%). The expression of the nv-gvp genes was monitored in N. vacuolatum cells cultivated in 20 or 13% salt media. Northern and Western analyses showed that despite the lack of gas vesicles in cells grown in 13% salt medium, the gvpACNOFGH gene cluster was transcribed and GvpA protein was synthesized, suggesting that the absence of gas vesicles is not due to a lack of transcription. PMID- 9211712 TI - Rhodospira trueperi gen. nov., spec. nov., a new phototrophic Proteobacterium of the alpha group. AB - A new phototrophic purple bacterium was isolated from a flat, laminated microbial mat in a salt marsh near Woods Hole, Mass., USA. The spiral-shaped bacterium was highly motile and had bipolar tufts of flagella and intracytoplasmic membranes of the vesicular type. The major photosynthetic pigments were identified as the carotenoid tetrahydrospirilloxanthin and bacteriochlorophyll b. The long wavelength in vivo absorption maximum of the bacteriochlorophyll was at 986 nm. The marine bacterium showed optimal growth in the presence of 2% NaCl. It utilized a number of organic substrates as carbon and energy sources and required vitamins and sulfide as a reduced sulfur source for growth. In the presence of sulfide, elemental sulfur globules were formed outside the cells. Elemental sulfur was not further oxidized to sulfate. The new isolate had a unique lipid and fatty acid composition, and according to the 16S rRNA gene sequence, it is most similar to Rhodospirillum rubrum. It is described as a new species and assigned to a new genus with the proposed name Rhodospira trueperi. PMID- 9211713 TI - Changes in the growth and enzyme level of Zymomonas mobilis under oxygen-limited conditions at low glucose concentration. AB - Zymomonas mobilis growing aerobically with 20 g glucose-1 (carbon-limited) in a chemostat exhibited an increase in both the molar growth yield (Yx/s) and the maximum molar growth yield (Yx/smax) and a decrease in both the specific substrate consumption rate (qs) and the maintenance energy consumption rate (me). Stepwise increase in the input oxygen partial pressure showed that anaerobic-to aerobic transitional adaptation occurred in four stages: anaerobic (0 mm HgO2), oxygen-limited (7.6- 230 mm HgO2), intermediate (273 mm HgO2), and oxygen excess (290 mm HgO2). The steady-state biomass concentration, Yx/s, and intracellular ATP content increased between oxygen partial pressures of 7.6 and 120 mm HgO2, accompanied by a decrease in the qs and the specific acid production rate. The membrane ATPase activity decreased with increasing oxygen partial pressure and reached its lowest levels at 273 mm HgO2, which was the highest input oxygen partial pressure where steady-state conditions were possible. Glucokinase, glucose-6-phosphate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, and alcohol dehydrogenase activities also decreased when the oxygen partial pressure was increased above 15 mm Hg, whereas pyruvate decarboxylase was unaffected by aeration. Growth inhibition at 290 mm HgO2 was characterised by a drastic reduction in the pyruvate kinase activity and a collapse in the intracellular ATP pool. The growth and enzyme data suggest that at low glucose concentrations and oxygen-limited conditions, the increase in biomass yields is a reflection of a redirection of ATP usage rather than a net increase in energy production. PMID- 9211714 TI - The sae locus of Staphylococcus aureus controls exoprotein synthesis at the transcriptional level. AB - Agr and sar are known regulatory loci of Staphylococcus aureus that control the production of several extracellular and cell-wall-associated proteins. A pleiotropic insertional mutation in S. aureus, designated sae, that leads to the production of drastically diminished levels of alpha- and beta-hemolysins and coagulase and slightly reduced levels of protein A has been described. The study of the expression of the genes coding for these exoproteins in the sae::Tn551 mutant (carried out in this work by Northern blot analyses) revealed that the genes for alpha- and beta-hemolysins (hla and hlb) and coagulase (coa) are not transcribed and that the gene for protein A (spa) is transcribed at a somewhat reduced level. These results indicate that the sae locus regulates these exoprotein genes at the transcriptional level. Northern blot analyses also show that the sae mutation does not affect the expression of agr or sar regulatory loci. An sae::Tn551 agr::tetM double mutant has been phenotypically characterized as producing reduced or null levels of alpha-, beta-, and delta-hemolysins, coagulase, and high levels of protein A. Northern blot analyses carried out in this work with the double mutant revealed that hla, hlb, hld, and coa genes are not transcribed, while spa is transcribed at high levels. The fact that coa is not expressed in the sae agr mutant, as in the sae parental strain, while spa is expressed at the high levels characteristic of the agr parental strain, suggests that sae and agr interact in a complex way in the control of the expression of the genes of several exoproteins. PMID- 9211715 TI - Properties and primary structure of a thermostable L-malate dehydrogenase from Archaeoglobus fulgidus. AB - A thermostable l-malate dehydrogenase from the hyperthermophilic sulfate-reducing archaeon Archaeoglobus fulgidus was isolated and characterized, and its gene was cloned and sequenced. The enzyme is a homodimer with a molecular mass of 70 kDa and catalyzes preferentially the reduction of oxaloacetic acid with NADH. A. fulgidus L-malate dehydrogenase was stable for 5 h at 90 degrees C, and the half life at 101 degrees C was 80 min. Thus, A. fulgidus L-malate dehydrogenase is the most thermostable L-malate dehydrogenase characterized to date. Addition of K2HPO4 (1 M) increased the thermal stability by 40%. The primary structure shows a high similarity to L-lactate dehydrogenase from Thermotoga maritima and gram positive bacteria, and to L-malate dehydrogenase from the archaeon Haloarcula marismortui and other L-lactate-dehydrogenase-like L-malate dehydrogenases. PMID- 9211716 TI - Adaptive response of Haloferax mediterranei to low concentrations of NaCl (< 20%) in the growth medium. AB - Halobacteria require 20-25% NaCl for optimal growth and lyse when the salt concentration falls below 10%. The response of Haloferax mediterranei cells to low concentrations of NaCl (< 20%) in the medium was studied. The cells adapted to and grew in concentrations of NaCl as low as 10% and survived in concentrations lower than 5%. The cells synthesised a red pigment, bacterioruberin, in response to stress caused by a low concentration of NaCl (< 20%). PMID- 9211717 TI - Swelling-activated organic osmolyte channels. PMID- 9211718 TI - Mutations in the M4 domain of the Torpedo californica nicotinic acetylcholine receptor alter channel opening and closing. AB - We studied the functional effects of single amino acid substitutions in the postulated M4 transmembrane domains of Torpedo californica nicotinic acetylcholine receptors (nAChRs) expressed in Xenopus oocytes at the single channel level. At low ACh concentrations and cold temperatures, the replacement of wild-type alpha418Cys residues with the large, hydrophobic amino acids tryptophan or phenylalanine increased mean open times 26-fold and 3-fold, respectively. The mutation of a homologous cysteine in the beta subunit (beta447Trp) had similar but smaller effects on mean open time. Coexpression of alpha418Trp and beta447Trp had the largest effect on channel open time, increasing mean open time 58-fold. No changes in conductance or ion selectivity were detected for any of the single subunit amino acid substitutions tested. However, the coexpression of the alpha418Trp and beta447Trp mutated subunits also produced channels with at least two additional conductance levels. Block by acetylcholine was apparent in the current records from alpha418Trp mutants. Burst analysis of the alpha418Trp mutations showed an increase in the channel open probability, due to a decrease in the apparent channel closing rate and a probable increase in the effective opening rate. Our results show that modifications in the primary structure of the alpha- and beta subunit M4 domain, which are postulated to be at the lipid-protein interface, can significantly alter channel gating, and that mutations in multiple subunits act additively to increase channel open time. PMID- 9211719 TI - Caco-2 cell line used as an in vitro model to study cadmium accumulation in intestinal epithelial cells. AB - 109Cd uptake was studied using the highly differentiated TC7 clone of Caco-2 cells as a model of human enterocyte function. Intracellular accumulation of 0.3 microM 109Cd involved a rapid and a slow uptake phase, which resulted in complete equilibration (t(1/2) = 17.3 +/- 1.3 min) with an apparent in-to-out distribution ratio (alphae) of 11.6 +/- 0.8. The amplitude of the rapid phase (U0) and the rate of the slow phase (V) were similarly reduced in the less differentiated PF11 clone, but comparable alphae values were observed at equilibrium. In both clones, the t(1/2) and alphae values increased and decreased, respectively, upon addition of unlabeled Cd to the uptake media. In TC7 cells, 109Cd uptake at 1 min (U1) was unaffected by Ca concentrations four order of magnitude in excess, but both U0 and V demonstrated similar sensitivities to unlabeled Cd, Zn and sulfhydryl reactive agents. Only U0 disappeared when EDTA was present in the wash solutions. U1 showed saturation kinetics and the data were found compatible with a model assuming rapid initial Cd binding and transport through a unique transport protein (Km = 3.8 +/- 0.7 microM). Cd efflux kinetics demonstrated partial reversibility in EDTA-containing solutions, suggesting that the taken up Cd might be both tightly and loosely bound to intracellular binding sites. However, the displacement of 109Cd measured at 65 min failed to reveal this heterogeneity: the data were found compatible with a model equation assuming the presence of one class of high-capacity high-affinity binding sites. We conclude that a slow transport fast-intracellular binding mechanism of Cd uptake best accounts for these results and that Cd transport most likely involves a carrier-type of protein unrelated to Ca absorption. PMID- 9211720 TI - The renal cortical Na+/HCO3- cotransporter VI: the effect of chemical modification in cotransporter activity. AB - The Na+/HCO3- cotransporter is the main system that mediates bicarbonate removal out of the proximal tubule cell into the blood. We have previously partially purified this protein and showed that chemical modification of the alpha-amino groups by fluorescein isothiocyanate (FITC) inhibited the activity of the Na+/HCO3- cotransporter. The inhibition was prevented by the presence of Na and bicarbonate suggesting that this compound binds at or near the substrate transport sites of the cotransporter. We examined the effect of agents that modify the sulfhydryl group (dithiothreitol), carboxyl groups (n-n'dicyclohexyl carbodiimide) and tyrosine residues (p-nitrobenzene sulfonyl fluoride, n-acetyl imidazole and tetranitromethane) on the activity of the cotransporter to gain insight into the chemical residues which may be important for transport function. The sulfhydryl residues modifier, carboxyl group modifier, and tyrosine modifier significantly inhibited bicarbonate dependent 22Na uptake in basolateral membranes by 50-70% without altering the 22Na uptake in the presence of gluconate indicating that these agents directly affected the cotransporter without affecting diffusive sodium uptake. The effect of the tyrosine modifier n acetylimidazole was not prevented by the presence of Na and bicarbonate suggesting that the tyrosine residues are not at the substrate binding sites. To determine the presence and role of glycosylation on the Na+/HCO3- cotransporter protein, we examined the effects of different glycosidases (endoglycosidase F and H, N-glycosidase F, O-glycanase) on the cotransporter activity. All glycosidases caused a significant 50-80% inhibition of cotransporter activity. These data demonstrate that N-glycosylation as well as O-glycosylation are important for the function of the Na+/HCO3- cotransporter protein. Taken together, these results suggest that chemical modifiers of tyrosine, carboxyl and sulfhydryl groups as well as glycosylation are important for expression of full functional activity of the cotransporter. PMID- 9211721 TI - Differential effect of high extracellular Ca2+ on K+ and Cl- conductances in murine osteoclasts. AB - Effects of the extracellular Ca2+ concentration ([Ca2+]o) on whole cell membrane currents were examined in mouse osteoclastic cells generated from bone marrow/stromal cell coculture. The major resting conductance in the presence of 1 mm Ca2+ was mediated by a Ba2+-sensitive, inwardly rectifying K+ (IRK) current. A rise in -Ca2+-o (5-40 mM) inhibited the IRK current and activated an 4'4' diisothiocyano-2,2'-stilbenedisulfonate (DIDS)-sensitive, outwardly rectifying Cl (ORCl) current. The activation of the ORCl current developed slowly and needed higher [Ca2+]o than that required to inhibit the IRK current. The inhibition of the IRK current consisted of two components, initial and subsequent late phases. The initial inhibition was not affected by intracellular application of guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) or guanosine 5'-O-(2-thiodiphosphate) (GDPbetaS). The late inhibition, however, was enhanced by GTPgammaS and attenuated by GDPbetaS, suggesting that GTP-binding proteins mediate this inhibition. The activation of the ORCl current was suppressed by pretreatment with pertussis toxin, but not potentiated by GTPgammaS. An increase in intracellular Ca2+ level neither reduced the IRK current nor activated the ORCl current. Staurosporine, an inhibitor for protein kinase C, did not modulate the [Ca2+]o-induced changes in the IRK and ORCl conductances. These results suggest that high [Ca2+]o had a dual action on the membrane conductance of osteoclasts, an inhibition of an IRK conductance and an activation of an ORCl conductance. The two conductances modulated by [Ca2+]o may be involved in different phases of bone resorption because they differed in Ca2+ sensitivity, temporal patterns of changes and regulatory mechanisms. PMID- 9211722 TI - Ca2+-dependent K+ channels in bovine adrenal chromaffin cells are modulated by lipoxygenase metabolites of arachidonic acid. AB - Fatty acids play an important role in a variety of physiological processes including ion channel modulation and catecholamine release. Using patch-clamp techniques we show that arachidonic acid (AA) is converted to lipoxygenase metabolites (LOMs) to potentiate activity of the Ca2+ and voltage-dependent, large-conductance K+ channel (BK) in bovine adrenal medullary chromaffin cells (BAMCCs). AA and LOM potentiation of BK current and recovery from potentiation were unaffected by the nonhydrolyzable ATP analogue AMP-PNP, or by exclusion of nucleotides in excised patch recordings. Also, AA and LOM potentiation of BK channel activity in outside-out patches exposed to strong Ca2+ buffering ruled out cytoplasmic messengers or changes in intracellular Ca2+ levels as causative factors. Lipoxygenase inhibitor attenuated AA, but not LOM potentiation of BK activity in outside-out patches, indicating that lipoxygenase processing of AA is possible in excised membrane patches, possibly via a membrane associated lipoxygenase. AA and LOM release have been implicated in the mechanics of catecholamine secretion from BAMCCs. By limiting action potential duration and thus voltage-gated Ca2+ influx, fatty acid potentiation of BK current may serve an inhibitory feedback function in regulating secretion from BAMCCs. PMID- 9211723 TI - 1/f-Noise of open bacterial porin channels. AB - General diffusion pores and specific porin channels from outer membranes of gram negative bacteria were reconstituted into lipid bilayer membranes. The current noise of the channels was investigated for the different porins in the open state and in the ligand-induced closed state using fast Fourier transformation. The open channel noise exhibited 1/f-noise for frequencies up to 200 Hz. The 1/f noise was investigated using the Hooge formula (Hooge, Phys. Lett. 29A: 139-140 (1969)), and the Hooge parameter alpha was calculated for all bacterial porins used in this study. The 1/f-noise was in part caused by slow inactivation and activation of porin channels. However, when care was taken that during the noise measurement no opening or closing of porin channels occurred, the Hooge Parameter alpha was a meaningful number for a given channel. A linear relationship was observed between alpha and the single-channel conductance, g, of the different porins. This linear relation between single-channel conductance and the Hooge parameter alpha could be qualitatively explained by assuming that the passing of an ion through a bacterial porin channel is-to a certain extent-influenced by nonlinear effects between channel wall and passing ion. PMID- 9211724 TI - Interconverting gating modes of a nonselective cation channel from the tapeworm Echinococcus granulosus reconstituted on planar lipid bilayers. AB - A 107-pS (symmetrical 150 mM KCl), nonselective cation channel was reconstituted from a microsomal membrane fraction of the larval stage of the tapeworm Echinococcus granulosus. Most of the time, it displayed a high open probability (>>0.95) irrespective of either the applied voltage, Ca2+, Ba2+, or tetraethylammonium concentration. Nevertheless, in contrast with this "leaklike" behavior, less frequently this "all-the-time-open" channel reversibly entered two different kinetic modes. One of them was characterized by lower Po values and some voltage sensitivity (V(1/2) congruent with 129 mV, and an equilibrium constant for channel closing changing e-fold per 63-mV change) the kinetic analysis revealing that it resulted from the appearance of voltage-sensitivity in the mean closed times and a sixfold increase in the equilibrium constant for channel closing at 0 mV. The other mode was characterized by a very fast open close activity leading to poorly resolved current levels and a Po around 0.6-0.7 which, occasionally and in a voltage-sensitive manner, entered a long-lived nonconducting state. However, the rare nature of these mode-shifting transitions precluded a more detailed analysis of their kinetics. The conductive properties of the channel were not affected by these switches. Model gating alone does not seem to ensure any physiological role of this channel and, instead, some other channel changes must occur if this phenomenon were to be of regulatory importance in vivo. Thus, mode-shifting might constitute an alternative target for channel activity modulation also in tapeworms. PMID- 9211725 TI - Further comments on codon reassignment. PMID- 9211726 TI - Further comments on codon reassignment. Response. PMID- 9211727 TI - Alu repeats and human evolution. PMID- 9211728 TI - Alu repeats and human evolution. Response. PMID- 9211729 TI - Evidence for the early divergence of tryptophanyl- and tyrosyl-tRNA synthetases. AB - Each amino acid is attached to its cognate tRNA by a distinct aminoacyl-tRNA synthetase (aaRS). The conventional evolutionary view is that the modern complement of synthetases existed prior to the divergence of eubacteria and eukaryotes. Thus comparisons of prokaryotic and eukaryotic aminoacyl-tRNA synthetases of the same type (charging specificity) should show greater sequence similarities than comparisons between synthetases of different types-and this is almost always so. However, a recent study [Ribas de Pouplana L, Furgier M, Quinn CL, Schimmel P (1996) Proc Natl Acad Sci USA 93:166-170] suggested that tryptophanyl- (TrpRS) and tyrosyl-tRNA (TyrRS) synthetases of the Eucarya (eukaryotes) are more similar to each other than either is to counterparts in the Bacteria (eubacteria). Here, we reexamine the evolutionary relationships of TyrRS and TrpRS using a broader range of taxa, including new sequence data from the Archaea (archaebacteria) as well as species of Eucarya and Bacteria. Our results differ from those of Ribas de Pouplana et al.: All phylogenetic methods support the separate monophyly of TrpRS and TyrRS. We attribute this result to the inclusion of the archaeal data which might serve to reduce long branch effects possibly associated with eukaryotic TrpRS and TyrRS sequences. Furthermore, reciprocally rooted phylogenies of TrpRS and TyrRS sequences confirm the closer evolutionary relationship of Archaea to eukaryotes by placing the root of the universal tree in the Bacteria. PMID- 9211730 TI - The major histocompatability complex (MHC) contains conserved polymorphic genomic sequences that are shuffled by recombination to form ethnic-specific haplotypes. AB - The major histocompatibility complex (MHC) consists of polymorphic frozen blocks (PFBs) that are linked to form megabase haplotypes. These blocks consist of polymorphic sequences and define regions where recombination appears to be inhibited. We have been able to show, using a highly polymorphic sequence centromeric of HLA-B (within the beta block), that PFBs are conserved and contain specific insertions/deletions and substitutions that are the same for individuals with the same MHC haplotype but that differ between at least most different haplotypes. A sequence comparison between ethnic-specific haplotypes shows that these sequences have remained stable and predate the formation of these haplotypes. To determine whether the same conserved block has been involved in the generation of multiple haplotypes, we compared the block typing profiles of different ethnic specific haplotypes. Block typing profiles have previously been shown to be identical in individuals with the same MHC haplotype but, generally, to differ between different haplotypes. It was found that some PFBs are common to more than one haplotype, implying a common ancestry. Subsequently, haplotypes have been generated by the shuffling and exchange of these PFBs. The regions between these PFBs appear to permit the recombination sites and therefore could be expected to exhibit either low polymorphism or a localized "hotspot." PMID- 9211731 TI - Genetic characterization of populations of a de novo arisen sugar beet pest, Aubeonymus mariaefranciscae (Coleoptera, Curculionidae), by RAPD analysis. AB - The weevil Aubeonymus mariaefranciscae is an important pest of sugar beet crops in southern Spain that was first described as a new species in 1979. We have studied the genetic variability of this insect by RAPD analysis of 122 individuals using eight primers. The high resolution provided by random amplified polymorphic DNA (RAPDs), in combination with efficient genetic distance estimators, allowed a very detailed description of the ecology and evolution of the populations of this insect in the south of Spain. The results are compatible with a unique event of colonization, followed by the spreading of the weevil across the surrounding areas. A comparison of the results obtained with different genetic distance estimators is presented. RAPD is shown to be a powerful technique for reconstructing the phylogenetic history of insect populations, even if they have diverged recently, which seems to be the case for A. mariaefranciscae. PMID- 9211732 TI - Molecular analysis of the intergenic region of the duplicated Amy genes of Drosophila melanogaster and Drosophila teissieri. AB - The intergenic regions between the duplicated amylase coding regions (Amy) of D. melanogaster and D. teissieri were sequenced. Their lengths in D. melanogaster and D. teissieri were 4,536 bp and 4,621 bp, respectively. Since homology between the upstream regions of the two duplicated genes was found up to 450 bp from the initiation codon of the Amy genes, the ancestral Amy coding region duplicated together with at least 450 bp of the 5'-flanking region as one unit. Comparison of the regions between the two species revealed that the level of divergence was very heterogeneous. Although the mean level of the nucleotide difference in this region was 0.107, no nucleotide substitution was found in four subregions whose sizes were more than 100 bp. Since the probability of these four subregions being completely conserved between D. melanogaster and D. teissieri was very low, these subregions were considered to have relatively important roles in evolution. Large insertions and deletions were not observed in this region but small ones were observed all over the region except for an about 1-kb subregion. This 1-kb region corresponded to an open reading frame encoding a protein which had some sequence identity with the proteins of the serine protease inhibitor superfamily (serpin). Since we could find a transcript of this gene and the synonymous substitution rate was higher than the replacement substitution rate, we suggest that this gene encodes an active serpin in Drosophila. PMID- 9211733 TI - Genomic organization of the extracellular coding region of the human FGFR4 and FLT4 genes: evolution of the genes encoding receptor tyrosine kinases with immunoglobulin-like domains. AB - Receptor tyrosine kinases with five, seven, and three Ig-like domains in their extracellular region are grouped in subclasses IIIa, IIIb, and IIIc, respectively. Here, we describe the genomic organization of the extracellular coding region of the human FGFR4 (IIIc) and FLT4 (IIIb) genes and compare it to that of the human FGFR1(IIIc), KIT, and FMS (IIIa). The results show that while genes belonging to the same subclass have an identical exon/intron structure in their extracellular coding region-as they do in their intracellular coding region genes of related subclasses only have a similar exon/intron structure. These results strongly support the hypothesis that the genes of the three subclasses evolved from a common ancestor by duplications involving entire genes, already in pieces. Hypotheses on the origin of introns and on the difference in the number of extracellular Ig-like domains in the three gene subclasses are discussed. PMID- 9211734 TI - Analysis of donor splice sites in different eukaryotic organisms. AB - We present here a new algorithm for functional site analysis. It is based on four main assumptions: each variation of nucleotide composition makes a different contribution to the overall binding free energy of interaction between a functional site and another molecule; nonfunctioning site-like regions (pseudosites) are absent or rare in genomes; there may be errors in the sample of sites; and nucleotides of different site positions are considered to be mutually dependent. In this algorithm, the site set is divided into subsets, each described by a certain consensus. Donor splice sites of the human protein-coding genes were analyzed. Comparing the results with other methods of donor splice site prediction has demonstrated a more accurate prediction of consensus sequences AG/GU(A,G), G/GUnAG, /GU(A,G)AG, /GU(A,G)nGU, and G/GUA than is achieved by weight matrix and consensus (A,C)AG/GU(A,G)AGU with mismatches. The probability of the first type error, E1, for the obtained consensus set was about 0.05, and the probability of the second type error, E2, was 0.15. The analysis demonstrated that accuracy of the functional site prediction could be improved if one takes into account correlations between the site positions. The accuracy of prediction by using human consensus sequences was tested on sequences from different organisms. Some differences in consensus sequences for the plant Arabidopsis sp., the invertebrate Caenorhabditis sp., and the fungus Aspergillus sp. were revealed. For the yeast Saccharomyces sp. only one conservative consensus, /GUA(U,A,C)G(U,A,C), was revealed (E1 = 0.03, E2 = 0.03). Yeast is a very interesting model to use for analysis of molecular mechanisms of splicing. PMID- 9211735 TI - Evidence for multiple functional copies of the male sex-determining locus, Sry, in African murine rodents. AB - Southern hybridization data suggest that the male sex-determining locus, Sry, is often duplicated in rodents. Here we explore DNA sequence evolution of orthologous and paralogous copies of Sry isolated from six species of African murines. PCR amplification followed by direct sequencing revealed from two to four copies of Sry per species. All copies include a long open reading frame, with a stop codon that coincides closely with the stop codon of the house mouse, Mus musculus, a species known to have a single copy of Sry. A phylogenetic analysis suggests that there are at least seven paralogous copies of Sry in this group of rodents. Putative orthologues are identical; sequence divergence among putative paralogues ranges from 1 to 8% (excluding the CAG repeat), with much lower levels of divergence in the high-mobility group (HMG-box) region than in the C-terminal region. A high proportion of nucleotide substitutions in both regions result in amino-acid replacement. The long open reading frame, conserved HMG-box, and pattern of evolution of the putative paralogues suggest that they are functional. PMID- 9211736 TI - Artiodactyl interspersed DNA repeats in cetacean genomes. AB - Interspersed repeats that emerged at different evolutionary times are informative in mammalian phylogeny. Here we show that the ancient short interspersed elements (SINEs) ARE1 and ARE2 are abundantly present in the genomes of artiodactyls and cetaceans but not in other mammalian genomes. This supports the classification of the cetaceans with the artiodactyls by a shared character that is unlikely to be the result of convergence. PMID- 9211738 TI - Complete large subunit ribosomal RNA sequences from the heterokont algae Ochromonas danica, Nannochloropsis salina, and Tribonema aequale, and phylogenetic analysis. AB - The large subunit ribosomal RNA sequences from the heterokont algae Ochromonas danica, Nannochloropsis salina, and Tribonema aequale were determined. These sequences were combined with small subunit ribosomal RNA sequences in order to carry out a phylogenetic analysis based on neighbor-joining, maximum parsimony, and maximum likelihood methods. Our results indicate that heterokont fungi and heterokont algae each are monophyletic, and confirm that they together form a monophyletic group called "stramenopiles." Within the heterokont algae, the eustigmatophyte Nannochloropsis salina either clusters with the chrysophyte Ochromonas danica or forms a sister group to a cluster comprising the phaeophyte Scytosiphon lomentaria and the xanthophyte Tribonema aequale. The alveolates were identified as the closest relatives of the stramenopiles, but the exact order of divergence between the eukaryotic crown taxa could not be established with confidence. PMID- 9211737 TI - Evolution of the mitochondrial DNA control region in the mbuna (Cichlidae) species flock of Lake Malawi, East Africa. AB - Considerable controversy has surrounded the application of mitochondrial DNA data to reconstruction of evolutionary relationships among the endemic cichlids of Lake Malawi. Central to this debate has been the issue of whether lineage sorting is complete, and thus whether these data actually reflect species phylogeny, or simply gene genealogy. Review of all mtDNA control region sequences available for members of one monophyletic subset of this species flock, the Malawi rockfishes, or mbuna, strongly indicates that lineage sorting is incomplete: Character-based analyses of these sequences reconstruct gene, not species, interrelationships. Analysis of the pattern of nucleotide substitutions differentiating these mtDNA alleles suggests that pyrimidine residues undergo transition substitutions more often than do purines. Estimation of the magnitude of derived sequence differentiation in light of the reconstructed gene genealogy suggests that the mbuna may be of considerably more recent vintage than previous molecular characterizations have indicated. PMID- 9211739 TI - The sperm nuclear basic proteins (SNBPs) of the sponge Neofibularia nolitangere: implications for the molecular evolution of SNBPs. AB - We have isolated and characterized for the first time, the SNBPs from an organism (Neofibularia nolitangere) of the phylum Porifera (Sponges). We have shown that these proteins consist of histones which, as expected, exhibit an amino acid composition very similar to that of other eukaryotic histones. The finding of histones in the sperm of these primitive organisms provides support to the notion that histones (SNBPs of the histone, H, type) were the proteins present at the onset of SNBP evolution. In contrast, a discrete number of alternative SNBP types (protamine-like, PL; protamine, P, types) seem to have appeared later on in the course of evolution and are found in both protostomes and deuterostomes, most likely as a result of processes of parallel evolution. PMID- 9211740 TI - An estimate of divergence time of Parazoa and Eumetazoa and that of Cephalochordata and Vertebrata by aldolase and triose phosphate isomerase clocks. AB - Previously we suggested that four proteins including aldolase and triose phosphate isomerase (TPI) evolved with approximately constant rates over long periods covering the whole animal phyla. The constant rates of aldolase and TPI evolution were reexamined based on three different models for estimating evolutionary distances. It was shown that the evolutionary rates remain essentially unchanged in comparisons not only between different classes of vertebrates but also between vertebrates and arthropods and even between animals and plants, irrespective of the models used. Thus these enzymes might be useful molecular clocks for inferring divergence times of animal phyla. To know the divergence time of Parazoa and Eumetazoa and that of Cephalochordata and Vertebrata, the aldolase cDNAs from Ephydatia fluviatilis, a freshwater sponge, and the TPI cDNAs from Ephydatia fluviatilis and Branchiostoma belcheri, an amphioxus, have been cloned and sequenced. Comparisons of the deduced amino acid sequences of aldolase and TPI from the freshwater sponge with known sequences revealed that the Parazoa-Eumetazoa split occurred about 940 million years ago (Ma) as determined by the average of two proteins and three models. Similarly, the aldolase and TPI clocks suggest that vertebrates and amphioxus last shared a common ancestor around 700 Ma and they possibly diverged shortly after the divergence of deuterostomes and protostomes. PMID- 9211741 TI - A protein related to eucaryal and bacterial DNA-motor proteins in the hyperthermophilic archaeon Sulfolobus acidocaldarius. AB - We have isolated a new gene encoding a putative 103-kDa protein from the hyperthermophilic archaeon Sulfolobus acidocaldarius. Analysis of the deduced amino-acid sequence shows an extended central domain, predicted to form coiled coil structures, and two terminal domains that display purine NTPase motifs. These features are reminiscent of mechanochemical motor proteins which use the energy of ATP hydrolysis to move specific cellular components. Comparative analysis of the amino-acid sequence of the terminal domains and predicted structural organization of this putative purine NTPase show that it is related both to eucaryal proteins from the "SMC family" involved in the condensation of chromosomes and to several bacterial and eucaryal proteins involved in DNA recombination/repair. Further analyses revealed that these proteins are all members of the so called "UvrA-related NTP-binding proteins superfamily" and form a large subgroup of motor-like NTPases involved in different DNA processing mechanisms. The presence of such protein in Archaea, Bacteria, and Eucarya suggests an early origin of DNA-motor proteins that could have emerged and diversified by domain shuffling. PMID- 9211742 TI - Cw*1701 defines a divergent african HLA-C allelic lineage. AB - The complete sequence of a new HLA-C allele, Cw*1701, was determined from a South African Zulu individual. Unique features that distinguish Cw*1701 from other HLA C alleles include multiple point substitutions and an 18 nucleotide insertion in exon 5, which encodes the transmembrane domain. In a phylogenetic analysis of HLA C sequences, Cw*1701 forms a third, distinct allelic lineage. A comparison of the transmembrane domain of Cw*1701 with other HLA-B and -C alleles reveals that duplications and deletions have been common in the evolution of these loci. A polymerase chain reaction based typing method was used to determine the distribution of this unusual allele in human populations. In contrast to the other two lineages of HLA-C alleles, the Cw*17 lineage is found at high frequencies only in populations of African descent. In addition, the HLA-B/Cw*17 haplotype diversity is higher in Africa. PMID- 9211744 TI - Functional motifs in the IgH enhancer of the channel catfish. AB - The transcriptional enhancer (Emu3') within the Ig heavy chain (IgH) locus of the channel catfish differs from those found in mammalian IgH loci in both its location and structure. However, upon transfection into fish or mouse lymphocytes, it activates transcription to an extent equivalent to that of the mouse IgH intronic enhancer (Emu). Potential transcription factor binding motifs in Emu3' are more numerous than in mammalian IgH enhancers, and are dispersed over 1.6 kilobases. We transfected catfish and mouse lymphoid cells with reporters under the control of artificial promoters containing motifs from the catfish enhancer. We demonstrate that 9 of 11 octamer motifs identified in the catfish enhancer, representing five variations of the consensus octamer (ATGCAAAT), are functional in both a catfish B-cell line (1B10) and the mouse plasmacytoma J558L. Only those octamer variants in which one of the first four bases is altered are active. Clear species differences in the strengths of the variant octamer motifs were evident, and in catfish B cells the ATGtAAAT motif was over threefold more active than the consensus octamer. The one muA and two muB motifs in Emu3' do not contribute to transcriptional activation. These results suggest that the relative functional contributions of IgH enhancer motifs has changed significantly during vertebrate evolution. PMID- 9211743 TI - Characterization of cDNA clones encoding mouse proteinase 3 (myeloblastine) and cathepsin G. AB - Serine proteases are the most abundant granule constituents of several major hematopoietic cell lineages. Due to their high abundance and their strict tissue specificity they have become important phenotypic cell markers used for studies of various aspects of hematopietic cell development. Using a polymerase chain reaction (PCR)-based strategy for the isolation of trypsin-related serine proteases, we were able to isolate cDNAs for two of the major neutrophil and monocyte serine proteases in the mouse, cathepsin G and mouse protease 3 (myeloblastin). The internal PCR fragments were used as probes to screen a mouse mast cell cDNA library and a cDNA library originating from a mouse monocytic cell line (WEHI-274.1). Full-length cDNAs for mouse cathepsin G and proteinase 3 were isolated and their complete sequences were determined. Northern blot analysis revealed expression of cathepsin G in immature cells of the monocyte macrophage lineage but also in the connective tissue mast cell line MTC. Proteinase 3 was expressed in several cell lines of myelo-monocytic origin and in one B-cell line, but not in any of the other cell lines tested. The isolation of cDNAs for mouse cathepsin G and mouse proteinase 3, together with the previous characterization of the gene for mouse N-elastase, and the entire or partial amino acid sequences for porcine azurocidine, equine N-elastase and proteinase 3, rat, dog, and rabbit cathepsin Gs in evolutionary relatively distantly related mammalian species, indicates that these four members of the serine protease family have been maintained for more than 100 million years of mammalian evolution. This latter finding indicates a strong evolutionary pressure to maintain specific immune functions associated with these neutrophil and monocyte proteases. All amino acid positions of major importance for the cleavage site selection have also been fully conserved between mouse and human proteinase 3 and a few minor changes have occurred between mouse and human cathepsin G. PMID- 9211745 TI - Sequence-based association analysis of HLA class I and II alleles in Japanese supports conservation of common haplotypes. AB - Alleles of HLA-A, B, C, DRB1, DQB1, and DPB1 loci were fully determined in 117 healthy Japanese. A*2402, A*3303, A*1101, A*0201, B*4403, B*5201, Cw*0102, Cw*1403, Cw*0304, Cw*0702, Cw*0801, and Cw*1202 showed frequencies of over 10%. Multi-locus haplotype frequencies were estimated by the maximum likelihood method. Strength of association between C and B loci was comparable with that between DRB1 and DQB1 loci. Alleles unidentified by a serological method and having very similar nucleotide sequences (A2: A*0201, A*0206, A*0207, B61: B*4002, B*4006) were carried by different haplotypes. Several frequent five-locus haplotypes were identified including A*3303-Cw*1403-B*4403-DRB1(*)1302 DQB1(*)0604, and A*2402-Cw*1202-B*5201-DRB1(*)1502-DQB1(*)0601. These sequence based haplotypes corresponded to serology-based common haplotypes which have already been described in Japanese. These findings indicate that common HLA haplotypes consist of particular sets of HLA alleles and that these haplotypes have been conserved through recent human evolution. PMID- 9211746 TI - Identification of new HLA class I region genes by sample sequencing. AB - Although many human major histocompatibility genes have been identified, relatively few have been localized to the class I region. We searched for new class I region genes by sample sequencing, a process in which short stretches of random genomic sequence are generated from cosmids and then compared with sequences deposited in nucleotide databases. Four class I region cosmids were isolated for sample sequencing by screening a chromosome 6 specific cosmid library with probes derived from specific class I region genes or with overlapping class I region yeast artificial chromosomes. Cosmids were sonnicated to produce fragments of 0.5 - 1 kilobases, subcloned, and sequenced using an automated sequencer. Sequences were then compared with nucleotide sequences deposited in the GenBank databases using the BLASTN algorithm. A number of potential new class I region genes were identified, including a cDNA with similarity to the tre oncogene, the trans-activating factor SC1 (TCF19), and a member of the interferon inducible 1 - 8 gene family. These observations suggest that sample sequencing is an efficient method for identifying new class I region genes, which can be applied to other regions of the genome and to other species, and support previous observations that the class I region contains a variety of genes other than those encoding HLA antigens. PMID- 9211747 TI - Transport-associated proteins in Atlantic salmon (Salmo salar). AB - The major histocompatibility complex (Mhc) regions of mice, rats, and humans all contain a pair of related genes, TAP1 and TAP2, which encode members of a large superfamily of proteins of similar structure and function. A functional TAP1/TAP2 heterodimer is probably required for efficient presentation of antigens to CD8(+) T cells. This heterodimer resides in the membrane of the endoplasmic reticulum, and transports peptides from the cytoplasm into the endoplasmic reticulum lumen for binding to Mhc class I molecules. The TAP transporter demonstrates specificity for both peptide sequence and length, and in rats, allelic variation in the sequence of the transporter molecules results in differential ability to transport particular peptides. Here we report two expressed Sasa-TAP2 loci, both of which are polymorphic, as well as an expressed Sasa-TAP1 locus from Atlantic salmon. The Sasa-TAP2A locus has a genomic organization similar to the human TAP2 equivalent. PMID- 9211748 TI - Absence of the hemochromatosis gene Cys282Tyr mutation in three ethnic groups from Algeria (Mzab), Ethiopia, and Senegal. AB - A Celtic origin for hemochromatosis, a common genetic iron metabolism disorder, has been postulated for a long time. To check whether the two mutations recently identified in the HLA-class I candidate gene for this disease were found only in Caucasians, we examined their frequencies in individuals originating from Algeria, Ethiopia, and Senegal. The presumably disease-causing mutation, responsible for the Cys282Tyr substitution, was not found in any member of these ethnic groups, although it was shown to be highly prevalent in populations of European ancestry. This geographic distribution supports the previously suggested Celtic origin for the disease. In contrast, the mutation responsible for the His63Asp substitution is not restricted to European populations. Although absent in the Senegalese, it was found on about 9% of the chromosomes of the Central Ethiopians and Algerians (Mzab) genotyped for this study. This second mutation, which probably represents a common variant unrelated to hemochromatosis, thus appears to have occurred earlier than that responsible for the Cys282Tyr substitution. More detailed population studies are needed to provide information on the age of these two mutations and eventually show how the hemochromatosis causing mutation chronologically spread throughout Europe. PMID- 9211749 TI - Germline TCR-A restriction of immunoglobulin E responses to allergen. AB - Immunoglobulin E responses to known environmental antigens (allergens) may serve as a general model to investigate germline genetic restriction of the immune response. We have previously shown genetic linkage between IgE responses to major allergens and the T-cell receptor (TCR) A/D locus, but not to TCR-B, implying that elements in TCR A/D restrict the ability to react to specific antigens. We now show, in two sets of subjects from the same population, a strong allelic association between a VA8.1 polymorphism (VA8.1(*)2) and reactivity to Der p II, a major antigenic component of the house dust mite Dermatophagoides pteronyssinus. Association was also seen between Der p II IgE titres and HLA DRB1(*)1501 alleles. Reactivity to Der p II was confined to subjects who were positive for VA8.1(*)2 and HLA-DRB1(*)1501, demonstrating germline HLA-DR and TCR A interaction in restricting the response to exogenous antigen. PMID- 9211750 TI - Association of human NK cell surface receptors NKR-P1 and CD94 with Src-family protein kinases. AB - Human natural killer (NK) cells express on their surface several members of the C type lectin family such as NKR-P1, CD94, and NKG2 that are probably involved in recognition of target cells and delivery of signals modulating NK cell cytotoxicity. To elucidate the mechanisms involved in signaling via these receptors, we solubilized in vitro cultured human NK cells by a mild detergent, Brij-58, immunoprecipitated molecular complexes containing the NKR-P1 or CD94 molecules, respectively, by specific monoclonal antibodies, and performed in vitro kinase assays on the immunoprecipitates. Sodium dodecyl sulfate polyacrylamide gel electrophoresis, autoradiography, and phospho-amino acid analysis revealed the presence of in vitro tyrosine phosphorylated proteins that were subsequently identified by re-precipitation (and/or by western blotting) as the respective C-type lectin molecules and Src family kinases Lck, Lyn, and Fyn. The NKR-P1 and the CD94-containing complexes were independent of each other and both very large, as judged by Sepharose 4B gel chromatography. Crosslinking of NKR-P1 on the cell surface induced transient in vivo tyrosine phosphorylation of cellular protein substrates. These results indicate involvement of the associated Src-family kinases in signaling via the NKR-P1 and CD94 receptors. PMID- 9211752 TI - Predominant role of N-terminal residue of nonamer peptides in their binding to HLA-B* 5101 molecules. PMID- 9211751 TI - Analysis of genetic diversity at the DQA loci in African cattle: evidence for a BoLA-DQA3 locus. AB - We describe the development of a polymerase chain reaction (PCR)-based approach for analysis of genetic diversity at the DQA loci in African Bos indicus and Bos taurus cattle. This approach, equally effective in European and Asian cattle breeds, detects the presence or absence of DQA1 and most duplicated DQA2 genes. Nucleotide and predicted amino acid sequence analysis of the highly polymorphic second exons, in addition to analysis of the locus-specific and relatively non polymorphic transmembrane, cytoplasmic, and 3-prime untranslated regions, has provided evidence for considerable diversity between each of the duplicated DQA2 genes. Therefore, we propose the designation BoLA-DQA3 for the previously unpublished alleles at the second DQA2 locus. Fourteen distinct PCR restriction fragment length polymorphism (RFLP) patterns, each identifying families of alleles at three DQA loci, can be distinguished. Nucleotide sequence analysis of new PCR-RFLP patterns from 193 Kenyan Boran, Ethiopian Arsi (B. indicus), and Guinean N'Dama (B. taurus) cattle identified 13 DQA1 alleles within eight major allelic families, five DQA2 alleles within a single allelic family, and seven DQA3 alleles within three major allelic families. PMID- 9211754 TI - Cardiac single-photon emission tomography: is attenuation correction enough? PMID- 9211753 TI - RT-PCR cloning and characterization of mouse immunoglobulin variable domains with high affinity for HLA-DR antigens. PMID- 9211755 TI - Evaluation of thyroid nodules with technetium-99m tetrofosmin dual-phase scintigraphy. AB - Technetium-99m tetrofosmin, a lipophilic cationic complex molecule, was introduced for myocardial imaging. In some biodistribution studies it has also been reported to accumulate in the thyroid gland. Our objectives were to determine which thyroid nodules retain tetrofosmin and whether preoperative evaluation of malignancy is possible. Tetrofosmin scintigraphy was performed in 57 patients with a cold thyroid nodule on previously performed pertechnetate scintigraphy. All patients had undergone ultrasonography and sonographically guided fine-needle aspiration biopsy. The tetrofosmin scintigrams were obtained 5 min (early image) and 1 h (late image) after intravenous injection of 370 MBq. Only nodules that showed clear tracer retention after 1 h in comparison with retention at 5 min were classified as TETRO positive. Nodules without late retention were classified as TETRO negative. All patients underwent surgery and the histological results were compared with the results of tetrofosmin scintigraphy. Ten out of 11 patients with thyroid carcinoma (two pT1, three pT2, five pT4) were TETRO negative. One patient with papillary carcinoma (pT2) was TETRO positive. The mean nodular to thyroid tissue (N/T) ratio for the late scan was 1.0+/-0.20. There were 21 patients with thyroid adenomas (seven follicular, seven microfollicular and seven oxyphilic); 15 of these patients were TETRO positive and six TETRO negative. The mean N/T ratio for the late images was 1.34+/-0.41. All patients with degenerative goitre (24 cases) and the one patient with Hashimoto's disease were TETRO negative after 1 h and the N/T ratio was 0.92+/-0.12 on the late scan. Our results indicate that 99mTc-tetrofosmin scanning is of little value preoperatively in distinguishing thyroid carcinoma from other thyroid nodules. Tetrofosmin tends to demonstrate thyroid adenomas but does not have a routine role in the assessment of thyroid nodules. PMID- 9211756 TI - The predictive value of serum thyroglobulin in the follow-up of differentiated thyroid cancer. AB - A strict and careful strategy has to be adopted to cure thyroid cancer. Diagnostic iodine-131 whole-body scan (WBS) and serum thyroglobulin (Tg) are important tools to detect thyroid remnants after thyroidectomy and radioiodine therapy. The aim of this retrospective study was to compare the relative sensitivity of WBS and Tg in the detection of thyroid remnants or metastases and to evaluate the predictive value of Tg in the clinical and scintigraphic course of the disease. Ninety-three patients were followed up after total thyroidectomy and the administration 4-6 weeks later of an ablative dose of 100 or 150 mCi 131I. Eighty-five percent of the patients were free of regional or distant metastases. The follow-up scheme included clinical examination of the patient followed by WBS, Tg, thyroid-stimulating hormone and free thyroxine measurements performed 4 weeks after thyroxine withdrawal and the observance of a low-iodine diet for at least 1 week. WBS (+) patients received a 100- or 150-mCi therapeutic dose of 131I. All patients were further followed up in the same way every 6 months until both WBS and Tg became negative, and thereafter at 1-, 2- and 4-year intervals. Six months after the postoperative radioiodine treatment (first visit), the sensitivity of WBS and Tg was 87% and 26% respectively. Among patients who were WBS(+) at the first visit, 95% of those who were Tg(-) and 47% of those who were Tg(+) had become disease-free at a median of 4 years after surgery (chi2=13.6; P<0.05). Patients whose tests were both positive required more radioiodine to be cured (335+/-90 vs 250+/-95 mCi; P<0.05). Our data indicate that in early diagnosed thyroid cancer, serum Tg measured 6 months after the postoperative 131I ablative dose is less sensitive than WBS for the demonstration of persistence of residual thyroid tissue but provides predictive information on the disease course. WBS(+) and Tg(-) patients are cured earlier and with less radioiodine than those who remain Tg(+). PMID- 9211757 TI - Indium-111 pentetreotide single-photon emission tomography in patients with TSH secreting pituitary adenomas: correlation with the effect of a single administration of octreotide on serum TSH levels. AB - Few data are available on the visualization of somatostatin receptors in vivo in patients with thyrotropin (TSH)-secreting adenoma. We studied five patients with TSH-secreting adenomas using single-photon emission tomography (SPET) after administration of indium-111 pentetreotide. The intensity of 111In-pentetreotide uptake by the tumours was correlated with the degree of TSH suppression after a single administration of 100 microg octreotide s. c. Five patients (three women and two men) aged 27-46 years were investigated. Except for one patient with acromegaly, all had pure TSH-secreting tumours. One patient was previously untreated, while two had received octreotide, one antithyroid drugs, and one radioiodine. In all patients SPET demonstrated increased uptake of 111In pentetreotide by the pituitary adenoma. The target to non-target ratio (T/nT) of 111In-pentetreotide uptake was higher than 10 in three patients. Administration of 100 microg octreotide s. c. caused a significant reduction in TSH levels from 4.8+/-1.4 mU/l to a nadir of 3.1+/-1.1 mU/l after 6 h (P<<0.001 by ANOVA). Suppression of TSH secretion ranged from 30% to 60% of the baseline value. The T/nT ratio showed a trend toward a direct relationship with the degree of TSH inhibition after acute octreotide administration (r=0.67; P=NS). Our study showed that 111In-pentetreotide scan visualized somatostatin receptors in all five of the patients with TSH-secreting pituitary adenomas, confirming the frequent presence of somatostatin receptors in these rare tumours, even though the correlation with the TSH inhibition after a single administration of octreotide did not reach significance. PMID- 9211758 TI - Chronic complicated osteomyelitis of the appendicular skeleton: diagnosis with technetium-99m labelled monoclonal antigranulocyte antibody-immunoscintigraphy. AB - Chronic post-traumatic osteomyelitis (OM) represents a particular challenge for nuclear medicine and radiology since clinical and biochemical parameters are frequently unreliable. The aim of this study was to investigate the value of combined bone scan (BS) and immunoscintigraphy (IS) with technetium-99m labelled monoclonal antigranulocyte antibody (MAB) in patients with suspected chronic OM of the appendicular skeleton. Twenty-four patients (17 females and 7 males) with suspected chronic post-traumatic OM were evaluated with three-phase BS/99mTc-MAB IS. The final diagnosis was established by means of bone culture and histology in 19 cases and clinical follow-up in five cases. The studies were reviewed by two independent and experienced observers; the interobserver agreement was calculated by kappa statistics. The sensitivity, specificity and accuracy of BS alone were 92%, 18% and 58%, respectively. Combined BS/99mTc-MAB-IS had a sensitivity, specificity and accuracy of 84%, 72% and 79%, respectively. Of 24 studies, 11 were true-positive, two false-negative, eight true-negative and three false positive. Two patients presented with unexpected ectopic haematopoietic bone marrow in the appendicular skeleton that caused false-positive results. A high degree of interobserver agreement was found (kappa=0. 85). It is concluded that combined BS/99mTc-MAB-IS represents a very sensitive and reproducible method with an acceptable specificity for the investigation of chronic OM. Problems may occur in the differentiation of low-grade OM from aseptic inflammation. Another problem is ectopic marrow that may occur in the appendicular skeleton due to a chronic inflammatory stimulus. A former intramedullary intervention in the femur with displacement of haematopoietic marrow may also lead to an ectopic location. PMID- 9211759 TI - Steady-state captopril renography: continuous monitoring of the captopril-induced increase in 99mTc-MAG3 mean parenchymal transit time in renovascular hypertension. AB - Steady-state captopril renography (SSCR) is an original technique for assessing the captopril-induced increase in technetium-99m mercaptoacetyltriglycine (99mTc MAG3) mean parenchymal transit time (MPTT) in kidneys affected with functional renal artery stenosis (RAS). The steady-state parenchymal activity achieved by constant infusion of 99mTc-MAG3 is directly linked to the MPTT of the radiopharmaceutical. This steady-state parenchymal activity was continuously monitored from 15 min before to 60 min after a single dose of captopril in order to detect possible disruption of the steady state. SSCR was performed in 11 hypertensive patients with unilateral RAS and in two with RAS of a solitary kidney before renal revascularization (RR). In four of these patients, an additional SSCR was performed after RR. Of the ten patients whose hypertension was cured or improved by RR, one presented an uninterpretable SSCR and six presented a positive SSCR on the affected side. Control SSCR performed in four of these six patients was bilaterally negative. SSCR was also bilaterally negative in the three patients who showed no blood pressure response to RR. These preliminary results tend to indicate that, in spite of the stability of pre- and post-captopril hydration and data acquisition conditions allowed by this steady state technique, the sensitivity is lower than expected. However, the reason for the false-negative results does not seem to be inherent to SSCR. PMID- 9211760 TI - Non-invasive assessment of technetium-99m albumin transit time distribution in the pulmonary circulation by first-pass angiocardiography. AB - This study describes a non-invasive method for assessment of the lung transit time distribution of a tracer, using first-pass technetium-99m albumin angiocardiography and a model-free method of deconvolution. Ten patients received a first injection of 1 MBq kg-1 in the external jugular vein to position a gamma camera in the left anterior oblique position and two additional injections (5 MBq kg-1) to record first-pass angiocardiographic data. Right and left ventricular time-activity curves were derived from regions of interest every 0.5 s over a 1 min period. The left ventricular curve was deconvoluted by the right ventricular curve to obtain the lung transport function. The deconvolution procedure was based on a modified version of the Kalman filtering technique. The procedure was repeated at an interval of 30 min in eight patients. Two patients were re examined up to 2 years later. Skewness, kurtosis and relative dispersion of the distributions did not change over time. We also found that the distribution, once normalized by its first moment, was independent of isolated changes in heart rate or cardiac output. Comparison of curve shapes at an interval of 30 min by point by point analysis demonstrated the reproducibility of the technique. We conclude that computation of the pulmonary transit time distribution of 99mTc-albumin from a standard angiocardiography procedure by model-free deconvolution is reliable and reproducible over time. We suggest that it may be a valuable tool for the non invasive follow-up of the pulmonary circulation. PMID- 9211762 TI - Technetium-99m sestamibi leg scintigraphy for non-invasive assessment of propionyl-L-carnitine induced changes in skeletal muscle metabolism. AB - Carnitine derivatives, such as propionyl-l-carnitine (PLC), have been shown to improve walking distance in patients with obstructive peripheral artery disease (PAOD). The aim of this study was to ascertain whether technetium-99m sestamibi leg scintigraphy may be a useful tool in the evaluation of changes in skeletal muscle metabolism induced by chronic therapy with PLC. Twenty patients with clinical and instrumental evidence of PAOD were randomly assigned to a 3-month period of therapy with either PLC or placebo. Rest 99mTc-sestamibi leg scintigraphy and echo-Doppler sonography were performed on all subjects immediately before and upon completion of the treatment period. At the end of the protocol the following results were observed in patients who underwent PLC administration: (a) a significant increase in both thigh and calf 99mTc-sestamibi uptake, in comparison with baseline values (P<0.001); (b) the absence of statistically significant modifications of Doppler blood flow indices of the lower limbs. In conclusion, after chronic administration of PLC, a significant increment in skeletal muscle uptake of 99mTc-sestamibi was demonstrated without any apparent change in regional blood flow. This fact, if proven in further studies, may suggest a role for this tracer as a non-invasive probe of tissue bioenergetics. PMID- 9211761 TI - Effects of aging on the cerebral distribution of technetium-99m hexamethylpropylene amine oxime in healthy humans. AB - Some brain functions decline at a linear rate throughout adulthood. Others remain relatively stable until very late in the life cycle. This study characterized the effects of aging on the regional cerebral distribution of hexamethylpropylene amine oxime (HMPAO) in healthy human volunteers. The sample consisted of 26 men and 18 women with a mean age of 41.6+/-14.9 years (range: 19-73). Their past medical histories, physical examinations, and laboratory screening tests were normal. Single-photon emission tomography (SPET) scans of the brain were performed with a standardized acquisition and processing protocol on a triple headed camera equipped with fan beam collimators. A 3-D restorative filter and a correction for uniform attenuation were applied before the images were reinterpolated in planes parallel to the line connecting the frontal and occipital poles. Mean counts per pixel were measured in multiple regions of interest (ROIs) within each hemisphere by custom fitting a set of templates to the images. The mean activity in each ROI was compared with the mean activity per pixel in the whole brain. Regression analyses were used to relate the activity ratios to age with both linear and nonlinear models. The relative concentration of radioactivity decreased significantly with age in most, but not all, gray matter structures. It increased in the white matter regions. The nonlinear model of aging fit the data significantly better than a straight line did. Most of the changes with age occurred during young adulthood. No further changes were detectable after the onset of middle age. The median breakpoint age at which the rate of change became negligible was 36.6 years. Aging significantly affects the relative uptake of HMPAO in healthy humans. It decreases in many gray matter regions and increases in most white matter regions. However, the changes do not appear to be linear. Most seem to occur during young adulthood before people reach their late thirties. The distribution then appears to remain relatively stable throughout middle age. PMID- 9211763 TI - Nitrate administration to enhance the detection of myocardial viability by technetium-99m tetrofosmin single-photon emission tomography. AB - A comparison was performed between technetium-99m tetrofosmin myocardial perfusion tomography at baseline and after nitrate administration, using a 2-day protocol, and rest-reinjection thallium-201 single-photon emission tomography (SPET) studies in order to assess whether nitrates enhance the detection of viable myocardium with 99mTc-tetrofosmin. Fifteen patients with coronary artery disease, previous myocardial infarction and a left ventricular ejection fraction <40% underwent 201Tl rest-injection and 99mTc-tetrofosmin baseline postnitroglycerin (0.4 mg sublingually) SPET studies, within 48 h. Tomograms based on the three spatial planes were divided into 15 segments and regional tracer uptake was quantitatively analysed. Viability was defined as presence of tracer uptake >/=50% of peak activity on baseline studies or after reversibility. The percentage of peak activity of 99mTc-tetrofosmin at baseline correlated with that of 201Tl (r=0.82, P <0.001). On baseline 99mTc-tetrofosmin studies, 73 of the 225 segments that were analysed had <50% of peak activity. Fifteen percent of these segments showed reversibility after nitrate administration, with an increase in 99mTc-tetrofosmin uptake from 40%+/-9% to 57%+/-9% of peak activity (P=0.003). All reversible segments after nitrate administration had viability criteria on 201Tl studies, but 20 segments that were non-viable on 99mTc tetrofosmin studies were viable on 201Tl studies. Using a threshold value of >/=40% of peak activity, only seven segments remained non-viable on 99mTc tetrofosmin studies. Overall agreement between 99mTc-tetrofosmin with nitrates and 201Tl-reinjection regarding the presence of myocardial viability was 90%. Detection of myocardial viability with 99mTc-tetrofosmin was enhanced after nitrate administration, correlating with viability criteria observed on thallium studies. PMID- 9211764 TI - Technetium-99m sestamibi myocardial tomography based on dipyridamole echocardiography testing in hypertensive patients with chest pain. AB - The non-invasive diagnosis of coronary artery disease in hypertensives with chest pain is an important clinical concern because all exercise-dependent tests display limited feasibility and diagnostic accuracy; by contrast, dipyridamole echocardiography testing has been shown to have a similar feasibility and accuracy in hypertensive and normotensive subjects. The aim of this study was to evaluate the diagnostic capability of technetium-99m sestamibi tomography based on dipyridamole echocardiography testing in hypertensives with chest pain, and to compare the scintigraphic results with those of coronary angiography, exercise electrocardiography and dipyridamole echocardiography. Forty subjects with mild to moderate hypertension, chest pain and no previous myocardial infarction were submitted to 99mTc-sestamibi tomography (at rest and after high-dose dipyridamole echocardiography) and to exercise electrocardiography testing. At coronary angiography 22 patients (group A) had significant epicardial coronary artery disease (>/=70% stenosis of at least one major vessel) and 18 normal main coronary vessels (group B). Dipyridamole 99mTc-sestamibi imaging was positive in 21/22 patients of group A and in 5/18 of group B. Dipyridamole echocardiography was positive in 18/22 patients of group A and in 5/18 of group B. Exercise electrocardiography was positive in 15/22 patients of group A and in 11/18 of group B. Four out of five subjects in group B with positive results in all the tests showed a slow run-off of angiographic contrast medium, probably due to small-vessel disease. Significant epicardial coronary artery disease in hypertensives with chest pain is unlikely when dipyridamole 99mTc-sestamibi tomography is negative. When scintigraphy is positive, either epicardial coronary artery disease or a small-vessel disease condition is possible. The association of scintigraphy with dipyridamole echocardiography testing allows the assessment of contractile function and myocardial perfusion by a single pharmacological stress. PMID- 9211765 TI - Use of scanner characteristics in iterative image reconstruction for high resolution positron emission tomography studies of small animals. AB - The purpose of this work was to improve of the spatial resolution of a whole-body positron emission tomography (PET) system for experimental studies of small animals by incorporation of scanner characteristics into the process of iterative image reconstruction. The image-forming characteristics of the PET camera were characterized by a spatially variant line-spread function (LSF), which was determined from 49 activated copper-64 line sources positioned over a field of view (FOV) of 21.0 cm. This information was used to model the image degradation process. During the course of iterative image reconstruction, the forward projection of the estimated image was blurred with the LSF at each iteration step before the estimated projections were compared with the measured projections. The imaging characteristics of the high-resolution algorithm were investigated in phantom experiments. Moreover, imaging studies of a rat and two nude mice were performed to evaluate the imaging properties of our approach in vivo. The spatial resolution of the scanner perpendicular to the direction of projection could be approximated by a one-dimensional Gaussian-shaped LSF with a full-width at half maximum increasing from 6.5 mm at the centre to 6.7 mm at a radial distance of 10.5 cm. The incorporation of this blurring kernel into the iteration formula resulted in a significantly improved spatial resolution of about 3.9 mm over the examined FOV. As demonstrated by the phantom and the animal experiments, the high resolution algorithm not only led to a better contrast resolution in the reconstructed emission scans but also improved the accuracy for quantitating activity concentrations in small tissue structures without leading to an amplification of image noise or image mottle. The presented data-handling strategy incorporates the image restoration step directly into the process of algebraic image reconstruction and obviates the need for ill-conditioned "deconvolution" procedures to be performed on the projections or on the reconstructed image. In our experience, the proposed algorithm is of special interest in experimental studies of small animals. PMID- 9211766 TI - Brain glucose utilization in systemic lupus erythematosus with neuropsychiatric symptoms: a controlled positron emission tomography study. AB - In contrast to morphological imaging [such as magnetic resonance imaging (MRI) or computed tomography], functional imaging may be of advantage in the detection of brain abnormalities in cases of neuropsychiatric systemic lupus erythematosus (SLE). Therefore, we studied 13 patients (aged 40+/-14 years, 11 female, 2 male) with neuropsychiatric SLE who met four of the American Rheumatism Association criteria for the classification of SLE. Ten clinically and neurologically healthy volunteers served as controls (aged 40+/-12 years, 5 female, 5 male). Both groups were investigated using fluorine-18-labelled fluorodeoxyglucose brain positron emission tomography (PET) and cranial MRI. The normal controls and 11 of the 13 patients showed normal MRI scans. However, PET scan was abnormal in all 13 SLE patients. Significant group-to-group differences in the glucose metabolic index (GMI=region of interest uptake/global uptake at the level of the basal ganglia and thalamus) were found in the parieto-occipital region on both sides: the GMI of the parieto-occipital region on the right side was 0.922+/-0.045 in patients and 1.066+/-0.081 in controls (P<<0.0001, Mann Whitney U test), while on the left side it was 0.892+/-0.060 in patients and 1. 034+/-0.051 in controls (P=0.0002). Parieto-occipital hypometabolism is a conspicuous finding in mainly MRI-negative neuropsychiatric SLE. As the parieto-occipital region is located at the boundary of blood supply of all three major arteries, it could be the most vulnerable zone of the cerebrum and may be affected at an early stage of the cerebrovascular disease. PMID- 9211767 TI - DOTATOC: a powerful new tool for receptor-mediated radionuclide therapy. AB - This study presents the first successful use of a peptidic vector, DOTATOC, labelled with the beta-emitting radioisotope yttrium-90, for the treatment of a patient with somatostatin receptor-positive abdominal metastases of a neuroendocrine carcinoma of unknown localization. Tumour response and symptomatic relief were achieved. In addition, the new substance DOTATOC was labelled with the diagnostic chemical analogue indium-111 and studied in three patients with histopathologically verified neuroendocrine abdominal tumours for its diagnostic sensitivity and compared with the commercially available OctreoScan. In all patients the kidney-to-tumour uptake ratio (in counts per pixel) was on average 1. 9-fold lower with 111In-DOTATOC than with OctreoScan. DOTATOC could be a potential new diagnostic and therapeutic agent in the management of neuroendocrine tumours. PMID- 9211768 TI - A clinical perspective of accelerated statistical reconstruction. AB - Although the potential benefits of maximum likelihood reconstruction have been recognised for many years, the technique has only recently found widespread popularity in clinical practice. Factors which have contributed to the wider acceptance include improved models for the emission process, better understanding of the properties of the algorithm and, not least, the practicality of application with the development of acceleration schemes and the improved speed of computers. The objective in this article is to present a framework for applying maximum likelihood reconstruction for a wide range of clinically based problems. The article draws particularly on the experience of the three authors in applying an acceleration scheme involving use of ordered subsets to a range of applications. The potential advantages of statistical reconstruction techniques include: (a) the ability to better model the emission and detection process, in order to make the reconstruction converge to a quantitative image, (b) the inclusion of a statistical noise model which results in better noise characteristics, and (c) the possibility to incorporate prior knowledge about the distribution being imaged. The great flexibility in adapting the reconstruction for a specific model results in these techniques having wide applicability to problems in clinical nuclear medicine. PMID- 9211769 TI - Nuclear medicine techniques for the study of breast cancer. PMID- 9211770 TI - Injection-associated pain in femoral arteriography: a European multicenter study comparing safety, tolerability, and efficacy of iodixanol and iopromide. AB - PURPOSE: To evaluate injection-associated pain, safety, and efficacy with the isotonic contrast medium iodixanol (Visipaque 270 mg I/ml) compared with iopromide (Ultravist 300 mg I/ml) in femoral arteriography. METHODS: A multicenter, double-blind, randomized, parallel-group clinical investigation was carried out in 54 hospitals in Europe. Of the patients evaluated, 1225 received iodixanol and 1227 iopromide in conventional and/or digital subtraction angiography. RESULTS: The iodixanol group reported statistically significantly less injection-associated pain (0.9%) than the iopromide group (9.5%) (p << 0.001). Further, 4.1% in the iodixanol group experienced pain and/or severe heat sensation vs 19. 8% in the iopromide group (p << 0.001). In the iodixanol group, 1.8% of the patients experienced contrast-related adverse events vs 2.4% in the iopromide group (p = NS). Overall diagnostic information was optimal for 94.1% in the iodixanol group and 95.3% in the iopromide group (p = NS). CONCLUSIONS: Iodixanol 270 mg I/ml causes significantly less injection-associated pain during femoral arteriography and is as safe and efficacious as iopromide 300 mg I/ml. PMID- 9211771 TI - Patterns of recurrent disease after recanalization of femoropopliteal artery occlusions. AB - PURPOSE: In this prospective study we investigated the site, occurrence, and development of stenoses and occlusions following recanalization of superficial femoral artery occlusions. METHODS: Recanalization of an occluded femoropopliteal artery was attempted in 62 patients. Follow-up examinations included clinical examination and color-flow duplex scanning at regular intervals. Arteriography was used to determine the localization of the recurrent disease relative to the initially occluded segment. RESULTS: During a mean follow-up of 23 months (range 0-69 months) 14 high-grade restenoses, indicated by a peak systolic velocity ratio >> 3.0, were detected by color-flow duplex scanning. Occlusion of the treated segment occurred in 11 patients. The cumulative 3-year primary patency rate for high-grade restenoses and occlusions combined was 44% (SE 9%). By arteriographic examination the site of restenosis was localized in the distal half of the treated vessel segment in 16 of 21 cases. CONCLUSION: Most restenoses and occlusions occurred during the first year and most disease developed at the previous intervention site. The site of restenosis is more frequently in the distal part of the initially treated segment, a finding that may have therapeutic implications. PMID- 9211772 TI - Percutaneous transluminal rotational atherectomy in the treatment of peripheral vascular disease using a transluminal endatherectomy catheter (TEC): initial results and angiographic follow-Up. AB - PURPOSE: To evaluate the clinical results of percutaneous transluminal rotational atherectomy in the treatment of peripheral vascular disease. METHODS: Rotational atherectomy was performed in 39 patients aged 39-87 years (mean 66.6 years). A total of 71 lesions (43 stenoses and 28 occlusions) were treated in 40 limbs. Additional balloon angioplasty was required in 54% of lesions. Fifteen patients (37.5%) presented in Fontaine stage II, 10 patients (25%) in Fontaine stage III and 15 patients (37.5%) in Fontaine stage IV. Rotational atherectomy at 750 rpm was carried out over a 0.014-inch guidewire with continuous aspiration into a vacuum bottle. Follow-up angiography and color flow Doppler examinations were performed in 22 patients (23 limbs) after a mean period of 6 months (range 2-14 months). RESULTS: There was one primary technical failure. In 36 of 40 lesions there was a good angiographic result with residual stenoses in less than 30%. In 70 lesions treated by rotational atherectomy, however, 54% showed residual stenoses of 30%-50% and these cases required additional balloon angioplasty. The mean ankle-brachial index improved significantly (p << 0.001) from 0.49 before the procedure to 1.01 after the procedure. A single distal embolus, related to primary recanalization, occurred and there were two large inguinal hematomas. Cumulative clinical patency after 6 months was 83.8% and cumulative angiographic patency after 6 months was 79.1%. CONCLUSION: Percutaneous rotational atherectomy is a promising approach for the treatment of chronic peripheral vascular disease. Further prospective, randomized studies are necessary to compare percutaneous transluminal angioplasty with this new technical approach. PMID- 9211774 TI - The role of interventional radiology in the management of intra- and extra peritoneal leakage in patients who have undergone continent urinary diversion. AB - PURPOSE: To assess how radiologic intervention altered the hospital course of patients undergoing continent urinary diversion. METHODS: Thirty-seven consecutive patients with bladder cancer invading the muscular layer were treated with total cystectomy and construction of a continent urinary reservoir. Eleven of 37 patients suffered early and late anastomotic leakage; six had prolonged extraperitoneal leakage at the urethroenteric anastomosis, three had prolonged intraperitoneal pouch leaks, and two had delayed ureteroenteric leaks. Seven of these patients required radiologic intervention. RESULTS: Intervention in the form of drainage catheter manipulation (n = 4), percutaneous nephrostomy (n = 4), or ureteral stent placement (n = 2) resulted in cessation of leakage without surgical intervention in all seven patients. Intraperitoneal pouch leaks were more difficult to control than extraperitoneal leakage and required longer drainage intervals. CONCLUSION: Interventional radiologic procedures played a key role in the management of continent urinary diversion complications, obviating the need for repeat surgical intervention in all instances. PMID- 9211773 TI - Venous malformations: sclerotherapy with a mixture of ethanol and lipiodol. AB - PURPOSE: To evaluate the usefulness of a mixture of absolute ethanol and lipiodol in the management of venous malformations. METHODS: Percutaneous sclerotherapy was performed with a mixture of absolute ethanol and lipiodol (9:1) in 17 patients with venous malformations, once in 12 patients, twice in 5. The therapeutic efficacy was evaluated by pain reduction. Conventional radiographs (n = 15) and posttreatment magnetic resonance imaging (n = 5) were obtained for the follow-up evaluation. RESULTS: Sclerotherapy was successful in all but two patients. The therapeutic effect was excellent in two patients, good in seven, fair in five, and poor in one. Radiopacity of lipiodol was beneficial for monitoring the procedure rather than for follow-up evaluations. Areas with low signal-intensity strands were increased on T2-weighted images obtained after the sclerotherapy. CONCLUSION: Sclerotherapy with a mixture of ethanol and lipiodol is effective in treating venous malformations. PMID- 9211775 TI - Reduced systemic toxicity from superselective chemoembolization compared with systemic chemotherapy in patients with high-risk metastatic gestational trophoblastic disease. AB - PURPOSE: The efficacy of chemoembolization of primary and metastatic gestational trophoblastic neoplasms was studied. METHODS: Six female patients, 19-33 years old, with high-risk trophoblastic disease were subjected to one to five chemoembolizations in 3-week intervals. Three of the patients had metastases to the liver, 2 had local tumor extension to the pelvic wall, and all 5 had failed initial systemic chemotherapy. The sixth patient was treated for a trophoblastic remnant following surgical expression of a tubal pregnancy. For follow-up, beta hCG levels in urine and serum and dynamic or angio-computed tomograms were obtained in biweekly to 6-month intervals. RESULTS: Two of 3 patients with liver metastases are alive and free of disease 6 and 7 years after initial chemoembolization. The third is alive at 3 years but with evidence of recurrent disease. Two patients treated for locally invasive trophoblastic disease died 3 months and 4 years, respectively, after initial chemoembolization. One had a 2 1/2 -year remission. The patient treated for a trophoblastic remnant in the tube is alive and free of disease at 6-year follow-up. Hematologic toxicity occurred in only one. CONCLUSION: Selective chemoembolization in our small series of patients with high-risk trophoblastic disease was equally effective as results reported for multi-drug systemic chemotherapy but had markedly lower renal, liver, and hematologic toxicity. PMID- 9211776 TI - Posterior pelvic ring fractures: closed reduction and percutaneous CT-guided sacroiliac screw fixation. AB - PURPOSE: To assess the midterm results of closed reduction and percutaneous fixation (CRPF) with computed tomography (CT)-guided sacroiliac screw fixation in longitudinal posterior pelvic ring fractures. To document radiographic and CT follow-up patterns. METHODS: Thirteen patients with 15 fractures were treated. Eleven patients received a unilateral, two a bilateral, screw fixation. Twenty seven screws were implanted. Continuous on-table traction was used in six cases. Mean radiological follow-up was 13 months. RESULTS: Twenty-five (93%) screws were placed correctly. There was no impingement of screws on neurovascular structures. Union occurred in 12 (80%), delayed union in 2 (13%), and nonunion in 1 of 15 (7%) fractures. There was one screw breakage and two axial dislocations. CONCLUSION: Sacroiliac CRPF of longitudinal fractures of the posterior pelvic ring is technically simple, minimally invasive, well localized, and stable. It should be done by an interventional/surgical team. CT is an excellent guiding modality. Closed reduction may be a problem and succeeds best when performed as early as possible. PMID- 9211777 TI - MR-guided cholecystostomy: assessment of biplanar, real-time needle tracking in three pigs. AB - PURPOSE: To demonstrate the feasibility of magnetic resonance (MR)-guided cholecystostomy using active, real-time, biplanar MR tracking in animal experiments. METHODS: Experiments were performed on three fully anesthetized pigs in an interventional MR system (GE open). The gallbladder was displayed in two orthogonal planes using a heavily T2-weighted fast spin-echo sequence. These "cholangio roadmaps" were displayed on LCD monitors positioned in front of the interventionalist. A special coaxial MR-tracking needle, equipped with a small receive-only coil at its tip, was inserted percutaneously into the gallbladder under continuous, biplanar MR guidance. The MR-tracking sequence allowed sampling of the coil (needle tip) position every 120 msec. The position of the coil was projected onto the two orthogonal "cholangio roadmap" images. RESULTS: Successful insertion of the needle was confirmed by aspiration of bile from the gallbladder. The process of aspiration and subsequent instillation of Gd-DTPA into the gallbladder was documented with fast gradient-recalled echo imaging. CONCLUSION: Biplanar, active, real-time MR tracking in combination with "cholangio roadmaps" allows for cholecystostomies in an interventional MRI environment. PMID- 9211778 TI - Three major coronary artery-to-left ventricular shunts: report of three cases and review of literature. AB - Among the congenital coronary artery fistulas, diffuse fistulation into the left ventricular chamber, usually expressed in terms of a coronary artery-left ventricular shunt, is not as rare today as was previously thought. However, the origin of such a shunt from all three major coronary arteries is rare. This paper reports three cases of such an occurrence and presents the clinical features and management of this rare anomaly by analyzing 31 cases, including 28 from the literature. PMID- 9211779 TI - Inadvertent rupture of a composite vein graft by angioplasty. AB - The superiority of vein over polytetrafluoroethylene (PTFE) as a bypass conduit for grafts ending below the knee makes it the material of choice for this purpose. When insufficient long saphenous vein is available, lengths of arm vein may be used as a satisfactory alternative to make a composite graft. This may cause confusion in subsequent graft surveillance programs as the arm vein segment may show different characteristics from the remainder of the graft. We report a case where a stenosis developed in the arm vein segment of a bypass graft which subsequently ruptured during balloon angioplasty with formation of a false aneurysm. This was due to the balloon size being selected on the basis of the size of the long saphenous vein section of the graft instead of the arm vein segment. Full communication between surgeon and radiologist must include complete details of all materials used in bypass grafts in order to avoid potentially disastrous results from angioplasty. PMID- 9211780 TI - Small cystic insulinoma: value of arterial stimulation venous sampling. AB - Cystic insulinomas are rare, with only three cases having been reported in the literature. It is not difficult to determine the site of such neoplasms, as cystic insulinomas are usually 4-10 cm in diameter. We report a patient with a histologically confirmed cystic insulinoma. This case is unique because of the small size (1.3 cm) of the tumor. Arterial stimulation venous sampling was useful for localizing and distinguishing this tumor from other pancreatic lesions. PMID- 9211781 TI - Acute Budd-Chiari syndrome: treatment with transjugular intrahepatic portosystemic shunt. AB - The case of a 28-year-old man with acute Budd-Chiari syndrome due to veno occlusive disease is reported. Transjugular intrahepatic portosystemic shunt (TIPS) was performed after upper gastrointestinal endoscopy, duplex sonographic and abdominal computed tomographic examination, inferior cavogram with hepatic venous catheterization, and transvenous biopsy. A 10-mm parenchymal tract was created. The patient did well after the procedure; ascites resolved and liver function improved markedly. The shunt has remained patent up to now for 6 months. PMID- 9211782 TI - Successful repeat transcatheter ablation of a mediastinal parathyroid adenoma 6 years after alcohol embolization. AB - Recurrent hyperparathyroidism is rare following transcatheter ablation of mediastinal parathyroid adenomas. When it occurs it is usually early and resistant to further attempts at ablation. We present a patient with primary hyperparathyroidism in whom two surgical attempts at cure had been unsuccessful. Subsequently, a mediastinal adenoma was demonstrated angiographically and embolized with absolute alcohol. Hyperparathyroidism recurred 6 years later and the mediastinal adenoma was subsequently successfully ablated a second time by angiographic embolization with ionic contrast medium. PMID- 9211783 TI - Button self-retaining drainage catheter. AB - To help improve patient acceptance of long-term internal/external catheter access to the biliary tract in those with benign biliary obstruction, a simple design allows the catheter end to remain flush with the skin. It consists of a clothes button affixed to the drainage catheter with a wood screw after the catheter has been cut off at the skin exit. This button/screw device has been used successfully in 22 patients over the last 10 years; catheter exchanges were easily accomplished. PMID- 9211784 TI - Percutaneous removal of a nonopaque silastic catheter from the pulmonary artery in two premature infants. AB - A modified snare was made from a 0.016' guidewire and a 0.1-mm fishing string to remove a nonopaque Silastic catheter via a femoral vein approach in 2 premature infants at the 44th and 120th day of life, respectively. A foldover guidewire loop snare had failed in 1 infant before this technique was successfully applied. PMID- 9211785 TI - Primary sarcoma of the aortic wall. PMID- 9211786 TI - Hepatocellular carcinoma with intraatrial extension. PMID- 9211787 TI - Incidence of SUC-RTM telomeric repeated genes in brewing and wild wine strains of Saccharomyces. AB - When over-expressed, RTM yeast genes confer resistance to the toxicity of molasses. They are found in distiller's and baker's industrial yeasts in multiple copies, scattered on the telomeres and physically linked to the telomeric SUC genes. Because these genes are absent from some laboratory strains, we explored the genomes of other industrial yeasts (brewing strains) and wine wild strains. A collection of 47 wine yeast strains (S. cerevisiae and S. bayanus) and 15 brewing strains, lager, ale and possible ancestors (S. monacensis, S. paradoxus and S. carlsbergensis) were screened for the presence of RTM genes. Only three wine strains and all brewing strains proved to contain RTM sequences in different copy numbers. PCR and chromosome blotting confirm the presence of SUC sequences in tandem with RTM. Moreover, analysis of the entire S. cerevisiae genome sequence shows that three other, non-telomeric, genes related to RTM are scattered on different chromosomes. PMID- 9211788 TI - Influence of gene dosage and autoregulation of the regulatory genes INO2 and INO4 on inositol/choline-repressible gene transcription in the yeast Saccharomyces cerevisiae. AB - Expression of structural genes of phospholipid biosynthesis in yeast is mediated by the inositol/choline-responsive element (ICRE). ICRE-dependent gene activation, requiring the regulatory genes INO2 and INO4, is repressed in the presence of the phospholipid precursors inositol and choline. INO2 and, to a less extent, INO4 are positively autoregulated by functional ICRE sequences in the respective upstream regions. However, an INO2 allele devoid of its ICRE functionally complemented an ino2 mutation and completely restored inositol/choline regulation of Ino2p-dependent reporter genes. Low-level expression of INO2 and INO4 genes, each under control of the heterologous MET25 promoter, did not alter the regulatory pattern of target genes. Thus, upstream regions of INO2 and INO4 are not crucial for transcriptional control of ICRE dependent genes by inositol and choline. Interestingly, over-expression of INO2, but not of INO4, counteracted repression by phospholipid precursors. Possibly, a functional antagonism between INO2 and a negative regulator is the key event responsible for repression or de-repression. PMID- 9211789 TI - The VPS4 gene is involved in protein transport out of a yeast pre-vacuolar endosome-like compartment. AB - Four yeast mutants were isolated in a screen for dominant-negative vacuolar protein-sorting mutants, secreting a carboxypeptidase Y-invertase hybrid protein. In addition to defects in the sorting/transport of soluble vacuolar hydrolases, the mutants accumulated a pre-vacuolar endosome-like compartment. The mutant alleles causing the defects were identified as the members of the VPS4 gene locus, each harbouring single-point mutations leading to amino-acid exchanges at positions 233 (E233Q), 211 (E211 K), and 178 (G178D). These mutations all reside within a 200 amino-acid-long ATPase module, common to members of the AAA-protein family. The VPS4 gene product shows homology to the yeast Sec18p (50% similarity and 25% identity), which is involved in several vesicle-mediated protein transport steps and homotypic membrane fusion events. Disruption of the VPS4 gene leads to a recessive vacuolar protein-sorting phenotype. About 40% of newly synthesized CPY is secreted as the Golgi-modified p2CPY precursor form. Transport of secretory proteins to the plasma membrane is normal as demonstrated by the secretion of invertase and alpha-factor. The alpha-factor, however, is secreted as a partially processed precursor, caused by defects in late Golgi function. The vps4 mutants also exhibit defects in fluid-phase endocytosis, as demonstrated by the accumulation of Lucifer Yellow in a pre-vacuolar endosome-like compartment. Based on the pleiotropic phenotype of the vps4 mutants and on the sequence homology to NSF/Sec18p, we propose that the VPS4 gene product is required for efficient transport out of the pre-vacuolar endosome-like compartment. PMID- 9211790 TI - Caffeine-resistance in S. pombe: mutations in three novel caf genes increase caffeine tolerance and affect radiation sensitivity, fertility, and cell cycle. AB - Caffeine is a well known base analogue and is cytotoxic to both animal and yeast cells. There are two possible mechanisms by which yeast cells tolerate caffeine concentrations higher than normal, by mutation or by physiological adaptation. We have isolated novel caffeine-resistant mutants of S. pombe which define three distinct genes caf2, caf3 and caf4. These mutants achieved a level of caffeine resistance which is presumed to represent the upper limit attainable by mutation. The caf2-caf4 mutations, as well as the previously identified caf1 mutation, confer UV-sensitivity, caffeine-resistant UV repair, impaired fertility and sporulation, as well as a lengthened cell cycle. They are partially dominant for caffeine resistance and recessive for UV sensitivity. Some auxotrophic caf3-89 double mutants show drastically decreased caffeine resistance. The caf4 mutant is more resistant to gamma-radiation than wild-type cells and shows pH-sensitive growth. As each caf mutation can, individually, confer maximum caffeine resistance to the cells, all four genes are expected to operate in the same pathway. This pathway might also be responsible for the physiological adaptation since adaptation is lost in caf1-caf4 mutants. PMID- 9211791 TI - Mitochondrial ATP synthase subunit 9 is not required for viability of the petite negative yeast Kluyveromyces lactis. AB - Specific mutations in nuclear MGI genes encoding the alpha, beta and gamma subunits of the mitochondrial inner membrane F1-ATPase complex allow mitochondrial DNA (mtDNA) to be lost from K. lactis. In the absence of a mutation in any of these three nuclear genes, loss of mtDNA is lethal. These results imply that mtDNA encodes a gene that is essential. Likely candidates for such an essential role are the ATP6, 8 and 9 genes coding for proteins of the ATP synthase-F0 component. The present study removes ATP9 from contention as a vital mitochondrial gene because in a respiratory deficient mutant, Gly- 3. 9, lacking a nuclear mgi mutation, we have found that a rearrangement in mtDNA has deleted 22 amino acids from the carboxy terminus of the 75 amino-acid subunit-9 protein. Rearrangement in mtDNA has occurred by recombination at a 23-bp repeated sequence in the introns of the ATP9 and large ribosomal RNA (LSU) subunit genes. These two introns, of 394 (ATP9) and 410 (LSU) nucleotides, both belong to group 1. PMID- 9211793 TI - The wheat mitochondrial rps13 gene: RNA editing and co-transcription with the atp6 gene. AB - Northern analyses and reverse transcription-polymerase chain-reaction (RT-PCR) experiments, followed by PCR amplification product sequencing, were performed on total mitochondrial (mt) RNAs from wheat seedlings and tissue cultures. It was shown that the rps13 gene, which encodes ribosomal protein S13, and the atp6 gene, which encodes subunit 6 of the ATP synthase complex, were co-transcribed. However, rps13 transcripts were virtually undetectable in seedlings under conditions where atp6 transcripts appeared abundant. In addition, markedly higher steady state transcript levels were observed in tissue culture. Expression of the mitochondrial rps13 gene was confirmed by showing that its transcripts were edited. Slight differences between editing patterns of tissue-culture and whole plant transcripts were found. Taken together, these results suggest that in vitro culture could disturb the post-transcriptional regulation of gene expression. PMID- 9211792 TI - Additional copies of the mitochondrial Ef-Tu and aspartyl-tRNA synthetase genes can compensate for a mutation affecting the maturation of the mitochondrial tRNAAsp. AB - In an attempt to identify new nuclear genes involved in the synthesis and processing of mitochondrial tRNAs, we utilized a multicopy nuclear library to suppress the heat-sensitive phenotype of a Saccharomyces cerevisiae mitochondrial mutant strain. This strain (Ts 932) is defective in the 3'-end processing of the mitochondrial tRNAAsp transcript. The nuclear genes coding for the mitochondrial elongation factor Tuf M and for the mitochondrial aspartyl-tRNA synthetase have been found to restore the temperature-resistant phenotype and to correct the RNA processing defect. Suppression was effective even when the genes were present on a centromeric plasmid. PMID- 9211794 TI - Organization of chloroplast ribosomal RNA genes and in vitro self-splicing activity of the large subunit rRNA intron from the green alga Chlorella vulgaris C-27. AB - Sequencing of the rRNA gene (rrn) cluster of Chlorella vulgaris C-27 chloroplasts has revealed a striking organizational difference in comparison to another species of the same genus, Chlorella ellipsoidea C-87. The rrn23 gene in C. vulgaris is also split. However, the 815-bp intervening sequence in this gene has been identified as a group-I intron. An in vitro rrn23 transcript containing the entire intron and parts of flanking exon sequences is able to self-splice in vitro in the presence of GTP and Mg++. Accurate ligation of the exons has been confirmed by sequencing the cDNA of the spliced products. GTP labelling of total Chlorella RNA in vitro has revealed that the number of self-splicing RNAs present in Chlorella chloroplasts is limited compared to that found in other green algal species. PMID- 9211795 TI - Carbon regulation of the cuticle-degrading enzyme PR1 from Metarhizium anisopliae may involve a trans-acting DNA-binding protein CRR1, a functional equivalent of the Aspergillus nidulans CREA protein. AB - The pr1 gene of the entomopathogenic fungus Metarhizium anisopliae encodes a serine protease that is highly active towards the insect cuticle and whose synthesis is subject to both carbon and nitrogen repression. The pr1 promoter region was sequenced revealing the presence of putative CREA- and AREA-binding sites. In vitro bandshift experiments demonstrated that an Aspergillus nidulans GST-CREA fusion protein was capable of binding to two of the three putative CREA sites. Using a PCR-based strategy the M. anisopliae crr1 gene was identified; it encodes a putative C2H2-type DNA-binding protein with significant sequence similarity to A. nidulans CREA. Complementation experiments with an A. nidulans strain carrying creA204 demonstrated that CRR1 can partially substitute for CREA function. PMID- 9211796 TI - Truncated-gene reporter system for studying the regulation of manganese peroxidase expression. AB - The expression of manganese peroxidase (MnP) in nitrogen-limited cultures of Phanerochaete chrysosporium is regulated by Mn, heat shock (HS), and H2O2 at the level of gene transcription. We have constructed a homologous gene reporter system to further examine the regulation of two mnp genes, mnp1 and mnp2, encoding individual MnP isozymes. Internal deletions of 234 and 359 bp were made within the coding regions of the mnp1 and mnp2 genes, respectively. The truncated mnp genes were subcloned into the shuttle vector pOGI18, which includes the Schizophylum commune ade5 gene as a selectable marker, and transformed into a P. chrysosporium Ade1 auxotrophic mutant. Northern-blot analysis of purified Ade+ transformants demonstrated that both of the truncated mnp genes were regulated in a manner similar to the endogenous mnp genes with respect to nitrogen limitation and induction by Mn, HS, and H2O2. PMID- 9211797 TI - Disparate sequence characteristics of the Erysiphe graminis f.sp. hordei glyceraldehyde-3-phosphate dehydrogenase gene. AB - The Erysiphe graminis f.sp. hordei (Egh) glyceraldehyde-3-phosphate dehydrogenase (gpd) gene was isolated and characterized. It contains typical promoter elements and has three introns, one of which is positioned in the 5' untranslated region of the gene. The deduced amino-acid sequence has 87% similarity to gpd genes from other Ascomycete fungi. This is at the same level as previously estimated among these fungi. Comparison at the DNA level reveal similarities of only around 70%, which is 10% lower than previously reported. In an evolutionary tree based on the sequences from 18 fungal gpd genes, Egh falls into the group of Ascomycetes located at a basal position. The regulatory region of the Egh gpd gene has no homology to corresponding sequences in other filamentous Ascomycetes. Codon usage was determined for the four characterized Egh genes (tub2, Egh7, Egh16 and gpd) and found to be similar for all four genes. The results of the codon-usage analysis suggest that Egh is more flexible than other fungi in the choice of nucleotides at the wobble position. Codon-usage preferences in Egh and barley genes indicate a level of difference which may be exploited to discriminate between fungal and plant genes in sequence mixtures. The Egh gpd promoter appears to be superior to that of the Egh beta-tubulin gene (tub2) for driving the E. coli beta-glucuronidase (GUS) gene in transformation experiments. PMID- 9211798 TI - Profilin gene expression and regulation in a temperature-sensitive breast cancer cell line: tsFT101. AB - The temperature-sensitive mutant cells (tsFT101) derived from a mouse mammary carcinoma cell line, FM3A, become multinucleated at a non-permissive temperature of 39 degrees C. To further understand the molecular mechanism of such cytokinetic disturbance, we examined the expression of profilin, the main regulator of the transition of globular actin (G-actin) to filamentous actin (F actin). RT-PCR analysis of mouse profilin cDNA from tsFT101 showed a point mutation (177 A two head right arrow G) which was a wobble mutation causing no change in the encoded amino acid. The expression level of profilin mRNA was, however, diminished in cultured tsFT101 cells under non-permissive temperatures compared with wild-type FM3A cells in association with multinucleation. A stable transfection of profilin cDNA expression vector to tsFT101 cells prevented multinuclear cell formation when cultured at 39 degrees C. In contrast, antisense profilin cDNA expression vector did not alter multinuclear cell formation. The primary cause of the cytokinetic disturbance of tsFT101 cells may be due to the diminished level of profilin gene expression. PMID- 9211799 TI - Effects of myosin light chain kinase inhibitors on carbachol-activated nonselective cationic current in guinea-pig gastric myocytes. AB - The effects of myosin light chain kinase inhibitors on muscarinic stimulation activated nonselective cationic current (ICCh) in guinea-pig gastric antral myocytes were studied using the whole-cell patch-clamp technique. ICCh was induced by carbachol (CCh, 50 microM) at a holding potential of -30 mV or -60 mV. ML-7, a chemical inhibitor of myosin light chain kinase (MLCK), inhibited ICCh concentration dependently in a reversible manner (53 +/- 8.6% at 1 microM, mean +/- SE, n = 11). In addition, amplitudes of ICCh were only 37 +/- 2.7% of the daily control values following the addition of a peptide inhibitor of MLCK to the pipette solution. On the other hand, ML-7 had an inhibitory effect on voltage operated Ca2+ channel current. The peak value of Ba2+ current at 0 mV was reduced to 35 +/- 7.4% (n = 9) by 3 microM of ML-7. As ICCh is known to have an intracellular Ca2+ dependence, we tried to exclude the possibility that ML-7 inhibited ICCh indirectly via suppression of Ca2+ current and the similar inhibitory effects of ML-7 on ICCh were confirmed under the following conditions: (1) clamp of membrane potential at -60 mV; (2) clamp of intracellular [Ca2+] to 1 microM by 10 mM BAPTA; (3) pre-inhibition of Ca2+ channel by verapamil. Different from the effects on ICCh, ML-7 barely inhibited the same cationic current induced by guanosine 5'-O-(3-thiotriphosphate) (GTP[gammaS], 0.2 mM) in the pipette solution. These results suggest that a Ca2+/calmodulin-MLCK-dependent pathway can modulate the activation of ICCh in guinea-pig gastric antral myocytes. PMID- 9211800 TI - Elevation of plasma viscosity induces sustained NO-mediated dilation in the hamster cremaster microcirculation in vivo. AB - We studied whether a flow-independent increase of luminal wall shear stress (WSS) could dilate hamster arterioles in vivo and which endothelial mediators are potentially involved. To this end the plasma viscosity was elevated by exchanging blood for dextran-erythrocyte solution thereby augmenting WSS. Diameters of small and large arterioles as well as red blood cell velocities were measured before and after exchange of blood for solutions of identical haematocrit containing either high- (HMWD) or low-molecular weight dextran (LMWD). The potential role of endothelial autacoids was investigated by local application of the NO-synthase inhibitor NG-nitro-L-arginine (L-NNA), the inhibitor of cyclooxygenase, indomethacin (3 microM), or the K+-channel blocker, tetrabutylammonium (TBA, 0.1 mM) to assess the potential effects of EDHF. HMWD (n = 11 animals) increased plasma viscosity by 64 +/- 3% and dilated arterioles of all branching orders (A1 A4) significantly [by 24 +/- 3% (A1-A2) and 32 +/- 3% (A3-A4)]. This dilation compensated fully for the calculated initial increase of WSS. LMWD (n = 6) did not affect plasma viscosity or arteriolar diameters. Tissue treatment with L-NNA (30-300 microM, n = 12) substantially diminished the HMWD-induced dilation in small arterioles (A3-A4; to 13 +/- 3%; P<<0.05) and virtually abolished it in large ones (A1-A2). Consequently, the calculated WSS increased significantly in these arterioles (by 31 +/- 5%). TBA combined with L-NNA (n = 4) did not reduce further the remaining dilation. Indomethacin (n = 6) had no effect on HMWD induced dilation. We conclude that an increase of WSS induces a mainly NO mediated arteriolar dilation. This dilation occurs in all arteriolar branching orders and is of sufficient magnitude to compensate for the initial WSS-increase. Thus, any elevations of WSS fulfil the requirement for a signal to change diameter along the arteriolar tree in a coordinated manner. The fully compensating dilation which we observed indicates that WSS is a controlled variable. It does, however, raise questions as to its role as a continuous endothelial stimulus. PMID- 9211801 TI - Glibenclamide suppresses stretch-activated ANP secretion: involvements of K+ATP channels and L-type Ca2+ channel modulation. AB - The mechanism by which glibenclamide regulates mechanically activated atrial natriuretic peptide (ANP) secretion was investigated using isolated perfused rat atria. A reduction in atrial pressure from an experimentally imposed distending pressure stimulated the secretion of ANP and caused concomitant translocation of extracellular fluid (ECF) into the atrial lumen. The activation of ANP secretion and ECF translocation were closely correlated with atrial volume changes and the increase in ANP secretion was a function of the ECF translocation. Glibenclamide (1, 10, 100 microM), an ATP-sensitive K+ (K+ATP) channel blocker, had no effect on the basal secretion of ANP, suppressed the stimulation of stretch-activated ANP secretion in a dose-dependent manner, but not the translocation of the ECF. Glipizide (100 microM) and tolbutamide (100 microM), other K+ATP channel blockers, had similar effects to those of glibenclamide. Suppression by glibenclamide (100 microM) of the stretch-induced ANP secretion was not observed in atria that had been pretreated with pinacidil (200 microM), an ATP-sensitive K+ channel opener: pinacidil alone had no effect on ECF translocation and ANP secretion. Furthermore, blocking Ca2+ influx by using the Ca2+ channel blocker diltiazem (10 nM), or a Ca2+-depleted medium prevented the suppression of stretch induced ANP secretion by glibenclamide. In other experiments, atrial distension produced a slight membrane depolarization of cardiomyocytes; this was accentuated in the presence of glibenclamide. Furthermore, in single cardiomyocytes, glibenclamide increased the intracellular Ca2+ concentration ([Ca2+]i) in a dose dependent manner. From these results, we suggest that glibenclamide suppresses atrial release of ANP by blocking K+ATP channels and increasing Ca2+ influx and that the K+ATP channels are associated with the regulation of the mechanically activated ANP secretion from the atria. PMID- 9211802 TI - Partial recovery of in vivo function by improved incubation conditions of isolated renal proximal tubule. I. Change of amiloride-inhibitable K+ conductance. AB - Isolated microperfused rabbit renal proximal tubule S2 segments, if incubated in conventional substrate containing HCO3- Ringer solution, exhibit lower cell membrane potentials (Vb) and elevated intracellular Na+ concentrations ([Na]i) compared to rat tubules in vivo. Assuming that these and other differences reflect insufficient metabolic and/or hormonal stimulation of the cells, we have used microelectrode techniques to test whether improving substrate supply and applying norepinephrine (NE, to compensate for the missing nerve supply) reverts Vb and [Na]i to values observed in vivo. Application of D-glucose (5.5 mmol/l) and additional application of pyruvate, lactate, or L-alanine (each 10 mmol/l), or bathing the tubules in Dulbecco's modified Eagle's tissue culture medium (DMEM) significantly increased Vb and, whenever tested, reduced [Na]i as compared to substrate-free or D-glucose-containing control solution and these effects could be prevented - as tested in the case of pyruvate - by inhibition of the Na/K pump with ouabain. However, high concentrations of acetate, beta hydroxybutyrate, or L-glutamine had no significant effect. The largest effect was obtained with joint application of DMEM and NE (10 micromol/l) which increased Vb from -42.8 +/- 1.3 mV (SEM) to -55.3 +/- 2.5 mV (n = 11). Interestingly we noticed that under the latter conditions the Vb response to bath application of 1 mmol/l amiloride virtually disappeared, i.e. it changed from a depolarization of +14.6 +/- 1.4 mV (in D-glucose Ringer solution) to +0.6 +/- 0.7 mV (in DMEM plus NE) (n = 8), with some tubules showing even a small hyperpolarization. The latter implies partial restoration of the in vivo behaviour, since in experiments on rat proximal tubules in vivo amiloride regularly hyperpolarized the cells (by -3.4 +/ 0.76 mV, n = 5). Obviously under conventional in vitro conditions an amiloride inhibitable K+ conductance is activated which is inactive in vivo and also inactivates under improved conditions in vitro. In agreement with observations reported in the subsequent publication our results demonstrate that isolated proximal tubules undergo functional alterations which may be largely prevented by improved metabolic and stimulatory incubation conditions. PMID- 9211803 TI - Partial recovery of in vivo function by improved incubation conditions of isolated renal proximal tubule. II. Change of Na-HCO3 cotransport stoichiometry and of response to acetazolamide. AB - In the preceding publication we reported that some transport properties of proximal tubules perfused in vitro differ from those of tubules perfused in vivo, and that the in vivo function can be largely recuperated by improved metabolic substrate supply and stimulation with norepinephrine (NE). Since we have previously observed that the basolateral Na-HCO3-cotransporter operates with an overall stoichiometric ratio of q approximately 3 HCO3- :1 Na+ in vivo, but with q approximately 2 HCO3- :1 Na+ in vitro and that it responds differently in both cases to acetazolamide (ACZ), we have now tested whether the cotransporter can regain its in vivo function in vitro if the incubation conditions are improved. Cell membrane potentials (Vb) and cell pH (pHi) were measured with microelectrodes and microfluorimetric techniques on isolated S2 segments of rabbit proximal tubule and the instantaneous Vb response to a 2:1 reduction of bath HCO3- or Na+ concentration was determined. (DeltaVb)HCO3 and (deltaVb)Na averaged 13.1 +/- 0.9 mV (SEM) and 6.9 +/- 0.5 mV in D-glucose-containing control HCO3-Ringer solution and decreased respectively to 10.1 +/- 0.5 mV and 3.8 +/- 0.2 mV (n = 8) after incubation in tissue culture medium and NE (10(-5) mmol/l). These data imply that q increased from 1.9 to 2.7. Concomitantly the tubules became susceptible to ACZ (1 mmol/l), which reduced (deltaVb)HCO3 in control conditions only to 94.6 +/- 1.2% but under improved incubation conditions to 64.5 +/- 2.4%. As verified in voltage divider measurements the latter reduction was not caused by activation of a basolateral K+ conductance. The results indicate that improved incubation conditions can at least partially revert cotransport function towards that of the in vivo state. The effect of ACZ may be explained if in the improved state 1 CO32- + 1 HCO3- + 1 Na+ are cotransported, in which case inhibition of carbonic anhydrase (CA) may cause a CO32-/pH disequilibrium to develop in the basal labyrinth which impedes the cotransport. Under conventional incubation conditions, however, when only 2 HCO3- + 1 Na+ are cotransported no such disequilibrium should develop irrespective of whether CA is active or inhibited. PMID- 9211804 TI - Ochratoxin A disturbs pH homeostasis in the kidney: increases in pH and HCO3- in the tubules and vasa recta. AB - This study was designed to elucidate the effects of ochratoxin A (OTA) on pH homeostasis in the kidney. We measured pH in the proximal (PT) and the distal (DT) tubular fluid, the collecting duct urine (CD), the descending and the ascending vasa recta blood (VR), and the renal arterial blood (RA). OTA increased pH significantly in PT, DT, CD as well as in the descending and ascending VR, whereas pH in RA remained unchanged. We further determined CO2 tension (pCO2) and HCO3- in PT, CD as well as in the descending and ascending VR. OTA significantly increased HCO3- in PT, CD and the descending and ascending VR, with no changes in pCO2. Therefore, the increases in pH in PT, CD and the descending and ascending VR result from the increase in HCO3-. Our results suggest that OTA inhibits HCO3- reabsorption in the tubules, leading to the impairment of urinary acidification, and that OTA further leads to the disturbance of the acid-base state (alkalinization) in the interstitium in renal papilla. The impairment of urinary acidification may contribute to the disturbance of pH homeostasis in the renal papilla. The disturbance of pH homeostasis by OTA could be related to its nephrotoxicity. PMID- 9211805 TI - Early effects of denervation on sarcoplasmic reticulum properties of slow-twitch rat muscle fibres. AB - The Ca2+ release activity of the sarcoplasmic reticulum (SR) in chemically skinned single slow-twitch fibres from control, 2-day and 7-day denervated rat soleus muscle was studied. Histochemical fibre type composition of the whole muscle, electrophysiological properties and the Ca2+ sensitivity of tension development by single muscle fibres were also studied. All the data were correlated with contractile properties of the in vitro muscle. In the 2-day denervated muscle the SR Ca2+ capacity and the rate of Ca2+ uptake decreased from the control values of 0.384 +/- 0.030 micromol (mg fibre protein)-1 and 19.8 +/- 1.9 nmol min-1 (mg fibre protein)-1, respectively, to 0.210 +/- 0.016 micromol (mg fibre protein)-1 and 13.5 +/- 0.9 nmol min-1 (mg fibre protein)-1; the calculated amount of Ca2+ released upon stimulation by caffeine decreased from the control value of 0.148 to 0.078 micromol (mg fibre protein)-1. In the 7-day denervated muscle, the SR Ca2+ capacity and the rate of Ca2+ uptake increased to 0.517 +/- 0.06 micromol (mg fibre protein)-1 and 21.6 +/- 2.3 nmol min-1 (mg fibre protein)-1, respectively; the calculated amount of Ca2+ released increased to 0.217 micromol (mg fibre protein)-1. Both contraction time and tension of the isometric twitch decreased in 2-day denervated and increased in 7-day denervated muscles. Electrophysiological and histochemical changes, as well as changes in the Ca2+ sensitivity of the muscle fibres did not show any apparent correlation with mechanical changes. It is therefore concluded that the SR plays a prominent role in the early changes of contraction time and tension following denervation. PMID- 9211806 TI - P/Q-type calcium channels activate neighboring calcium-dependent potassium channels in mouse motor nerve terminals. AB - The identity of the voltage-dependent calcium channels (VDCC), which trigger the Ca2+-gated K+ currents (IK(Ca)) in mammalian motor nerve terminals, was investigated by means of perineurial recordings. The effects of Ca2+ chelators with different binding kinetics on the activation of IK(Ca) were also examined. The calcium channel blockers of the P/Q family, omega-agatoxin IVA (omega-Aga IVA) and funnel-web spider toxin (FTX), have been shown to exert a strong blocking effect on IK(Ca). In contrast, nitrendipine and omega-conotoxin GVIA (omega-CgTx) did not affect the Ca2+-activated K+ currents. The intracellular action of the fast Ca2+ buffers BAPTA and DM-BAPTA prevented the activation of the IK(Ca), while the slow Ca2+ buffer EGTA was ineffective at blocking it. These data indicate that P/Q-type VDCC mediate the Ca2+ influx which activates IK(Ca). The spatial association between Ca2+ and Ca2+-gated K+ channels is discussed, on the basis of the differential effects of the fast and slow Ca2+ chelators. PMID- 9211807 TI - Membrane currents in immature oocytes of the Rana perezi frog. AB - Immature oocytes of the Rana perezi frog were studied electrophysiologically to see if some of the unusual ionic channels found in Xenopus oocytes were also expressed in these cells. Growing oocytes showed a fairly linear current/voltage relationship (from -200 to +60 mV), whereas fully grown cells had several voltage dependent conductances. Depolarizing pulses elicited a potassium current blocked by tetraethylammonium (TEA) and two kinetically different Ca2+-dependent Cl- currents (ICl(Ca)), both sensitive to niflumic acid. ICl(Ca), which have not been previously observed in Rana immature oocytes, were also found in response to acetylcholine or rabbit serum superfusion or intracellular injection of Ca2+. In addition, three different Cl- currents were activated in these cells by hyperpolarization: (1) a transient inward current dependent on a critical intracellular Ca2+ concentration; (2) an inward rectifier Cl- current, which was Ca2+ independent; and (3) a high threshold (over -140 mV), slow Cl- current, blocked by several divalent cations, 4,4'-diisothiocyanatostilbene-2,2' disulphonic acid (DIDS) and 4-acetamido-4-isothiocyanatostilbene-2, 2' disulphonic acid (SITS). The presence of most of these infrequent currents in immature oocytes of several frogs and toads suggests that they are not merely the result of random genomic expression but a programmed decision, probably related to a definite functional role. PMID- 9211808 TI - Membrane responses to extracellular ATP in rat isolated white adipocytes. AB - We used whole-cell and perforated-patch voltage-clamp methods to study the membrane electrical properties of isolated rat epididymal and inguinal white adipocytes. We examined cells from both Sprague-Dawley and Zucker lean and Zucker obese (fa/fa) rats. A delayed-rectifier potassium current was present and similar in unstimulated white fat cells from all these sources. The potassium current activated rapidly with depolarization positive to about -30 mV and showed slow inactivation. Stimulation with extracellular ATP activated both hyperpolarizing and depolarizing conductances. ATP exposure also increased cell membrane capacitance by an average of 16%, suggesting that ATP activates exocytosis. Exposure to norepinephrine had little electrophysiological effect. We conclude that white adipocytes are very similar to brown adipocytes in their resting electrophysiological profile and in their responses to extracellular ATP. PMID- 9211809 TI - Assessment of evidence for K+-H+ exchange in isolated type-1 cells of neonatal rat carotid body. AB - Intracellular pH (pHi) was measured in enzymically isolated, neonatal rat carotid body type-1 cells, using the fluorophore carboxy-SNARF-1 (AM-loaded), and using the nigericin technique for in situ fluorescence calibration (nigericin is a membrane-soluble K+-H+ exchanger). In CO2/HCO3--free media, inhibiting Na+-H+ exchange produced a prompt fall of pHi (background acid-loading), the rate of which was reduced by raising the extracellular K+ concentration, [K+]o. pHi recovery from an intracellular acid or alkali load was also sensitive to changes of [K+]o. These results are similar to those of Wilding et al. (J Gen Physiol 100:593-608, 1992), who proposed the existence of an acid-loading, K+-H+ exchanger (KHE) in the type-1 cell. However, when nigericin was not used for post experimental calibration, and the superfusion system was flushed exhaustively with strong detergent, alcohol and distilled water, then background acid-loading was attenuated, and the K+o sensitivity of pHi insignificant. Background loading was increased again, and K+o sensitivity restored, when cells were monitored in a superfusion system which had previously been exposed to a single nigericin calibration protocol (followed by a short system wash with strong detergent and distilled water). We conclude that the previously reported expression of KHE in carotid body type-1 cells is an artefact caused by nigericin contamination. We have therefore quantified the pHi dependence of background loading in uncontaminated type-1 cells. We consider the possible implications of our work for reports of KHE in other cell types. PMID- 9211810 TI - Changes in baroreceptor vagal reflex performance in the developing rat. AB - Ontogenesis of both vagal control of heart rate and the baroreceptor vagal reflex were evaluated in rats at postnatal ages (P) of 5/6, 10, 15, 20, 25 and >>42 days anaesthetised with urethane (1.5 g/kg). Between P5/6 and P25 heart rate rose from 372 +/- 12 to 448 +/- 20 beats per minute and mean arterial pressure increased from 33.9 +/- 3.1 to 74.59 +/- 3.25 mm Hg (mean +/- SEM, n = 7 and 11 respectively). Cardiac vagal tone was absent at P10 but significant at P20 (P < 0.05) as revealed with atropine (0.5-1 mg/kg i.v.). Baroreceptor cardiac reflex sensitivity, tested with phenylephrine (10-50 microg/kg i.v.), was attenuated significantly in P10-20 rats compared with P5/6, P25 and mature animals. In P14 17 rats stimulation of neurones in either the solitary tract or ambiguual nuclei, by microinjection of L-glutamate (100-200 pmol), evoked an atropine-sensitive bradycardia indicating a functional integrity of central and peripheral efferent pathways mediating the baroreceptor reflex. Thus, the baroreceptor vagal reflex is functional in P5/6 rats but becomes attenuated between P10-P20, which is coincident with the maturational rise in arterial pressure. PMID- 9211811 TI - Blockade of nitric oxide decreases the renal vasodilatory effect of neuropeptide Y in the insulin-treated diabetic rat. AB - The effect of 50 days of streptozotocine-induced diabetes mellitus (blood glucose 20 mmol/l) on contraction and relaxation of isolated renal and intrarenal arteries in rats were examined. Strong and similar contractions were induced by potassium (60 mM), 5-hydroxytryptamine (5-HT) and endothelin-1 (ET-1) in renal and intrarenal arteries in diabetic and control rats. The vasodilatory reactivity, after precontraction with 5-HT, of neuropeptide Y (NPY) was similar to that of acetylcholine (ACh), calcitonin gene-related peptide (CGRP) and vasoactive intestinal polypeptide (VIP) and was similar in diabetic and control rats. The relaxing effect of NPY was decreased (40%) only in the diabetic group by blockade of nitric oxide synthase with NG-nitro-L-arginine methyl ester (10( 4) M) and by blockade (50%) of NPY with alpha-trinositol (10(-6) M). In conclusion, the present study showed that diabetes mellitus in the rat is associated with normal vasoconstrictive and vasodilatory capacities. However, the vasodilatory response to NPY was largely eliminated by blockade of nitric oxide synthesis only in the diabetic animals. This indicates that the vasodilatory effect of NPY in diabetes mellitus may be dependent on nitric oxide synthesis. PMID- 9211813 TI - Effects of reduced electrochemical Na+ gradient on contractility in skeletal muscle: role of the Na+-K+ pump. AB - Continued excitation of skeletal muscle may induce a combination of a low extracellular Na+ concentration ([Na+]o) and a high extracellular K+ concentration ([K+]o) in the T-tubular lumen, which may contribute to fatigue. Here, we examine the role of the Na+-K+ pump in the maintenance of contractility in isolated rat soleus muscles when the Na+, K+ gradients have been altered. When [Na+]o is lowered to 25 mM by substituting Na+ with choline, tetanic force is decreased to 30% of the control level after 60 min. Subsequent stimulation of the Na+-K+ pump with insulin or catecholamines induces a decrease in [Na+]i and hyperpolarization. This is associated with a force recovery to 80-90% of the control level which can be abolished by ouabain. This force recovery depends on hyperpolarization and is correlated to the decrease in -Na+-i (r = 0. 93; P<0.001). The inhibitory effect of a low -Na+-o on force development is considerably potentiated by increasing [K+]o. Again, stimulation of the Na+-K+ pump leads to rapid force recovery. The Na+-K+ pump has a large potential for rapid compensation of the excitation-induced rundown of Na+, K+ gradients and contributes, via its electrogenic effect, to the membrane potential. We conclude that these actions of the Na+-K+ pump are essential for the maintenance of excitability and contractile force. PMID- 9211812 TI - Calcium and magnesium transport in the cortical thick ascending limb of Henle's loop: influence of age and gender. AB - Previous studies from our laboratory have shown that Ca2+ and Mg2+ absorption in the mouse cortical thick ascending limb of Henle's loop (cTAL) is a passive, paracellular process driven by the transepithelial voltage. The passive permeability of the epithelium is enhanced by peptide hormones. The present study investigated whether divalent cation absorption in the cTAL is influenced by cell maturation and/or gender. For this purpose, mouse cTAL segments were microdissected from kidneys of female and male animals aged 4 and 8 weeks. The microdissected tubules were perfused in vitro at a luminal flow rate of 1.5 to 2.5 nl/min. Transepithelial Na+, Cl-, Ca2+ and Mg2+ net fluxes (JX, pmol.min-1.mm 1) were measured using electron microprobe analysis, and the transepithelial potential difference (PDte) was measured continuously. No differences were found in the PDte, JNa and JCl of the various animal groups but the transepithelial Ca2+ and Mg2+ transport capacity of the cTAL was higher in adults (8 weeks) than in young animals (4 weeks). Furthermore, irrespective of age, transepithelial Ca2+ net absorption was greater in male than in female animals. In contrast, the NaCl transport was maximal at 4 weeks in both genders. We conclude therefore that transepithelial divalent cation absorption in the mouse cTAL is an inductive process influenced by cell maturation and gender. The molecular basis of these inductions remains to be elucidated. PMID- 9211814 TI - Effect of intracellular pH on agonist-induced [Ca2+]i transients in HT29 cells. AB - In this study we examined the influence of intracellular pH (pHi) on agonist induced changes of intracellular Ca2+ activity ([Ca2+]i) in HT29 cells. pHi and [Ca2+]i were measured microspectrofluorimetrically using BCECF and fura-2, respectively. Buffers containing trimethylamine (TriMA), NH3/NH4+ and acetate were used to clamp pHi to defined values. The magnitudes of the peak and plateau of [Ca2+]i transients induced by carbachol (CCH, 10(-6) mol/l) were greatly enhanced by an acidic pHi and nearly abolished by an alkaline pHi. The relationship between pHi and the [Ca2+]i peak was nearly linear from pHi 7.0 to 7.8. This effect of pHi was also observed at higher CCH concentrations (10(-4 )and 10(-5) mol/l), at which the inhibitory effect of an alkaline pHi was more pronounced than the stimulatory effect of an acidic pHi. An acidic pHi shifted the CCH concentration/response curve to the left, whereas an alkaline pHi led to a rightward shift. The influence of pHi on [Ca2+]i transients induced by neurotensin (10(-8) mol/l) or ATP (5 x 10(-7) mol/l) was similar to its influence on those induced by CCH, but generally not as pronounced. Measurements of cellular inositol 1,4,5-trisphosphate (InsP3) showed no changes in response to acidification with acetate (20 mmol/l) or alkalinization with TriMA (20 mmol/l). The InsP3 increase induced by CCH was unaltered at an acidic pHi, but was augmented at an alkaline pHi. Confocal measurements of cell volume showed no significant changes induced by TriMA or acetate. Slow-whole-cell patch-clamp experiments showed no additional effect of CCH on the membrane voltage (Vm) measured after TriMA or acetate application. We conclude that pHi is a physiological modulator of hormonal effects in HT29 cells, as the [Ca2+]i responses to agonists were significantly changed at already slightly altered pHi. The measurements of InsP3, cell volume and Vm show that pHi must act distally to the InsP3 production, and not via changes of cell volume or Vm. PMID- 9211815 TI - Acidosis alters the phosphorylation of Ser16 and Thr17 of phospholamban in rat cardiac muscle. AB - The effect of acidosis on the phosphorylation of Ser16 and Thr17 of phospholamban in rat cardiac muscle has been investigated using phosphorylation-site-specific antibodies to this protein. Ventricular myocytes were stimulated at 0.5 Hz for 5 min, in either control (pH 7.4) or acid (pH 6.5) physiological salt solution, in the absence or presence of isoprenaline. Site-specific phosphorylation of phospholamban was determined by Western blotting. Acidosis reduced phosphorylation of Ser16 in the absence of isoprenaline, but did not alter the isoprenaline-induced phosphorylation of Ser16. In contrast, acidosis increased Thr17 phosphorylation in the absence and presence of isoprenaline. Buffering intracellular Ca2+ ([Ca2+]i) with BAPTA inhibited the increase in Thr17 phosphorylation during acidosis but had no effect on Ser16 phosphorylation. We conclude that acidosis can alter the phosphorylation of Ser16 and Thr17 by inhibition of protein kinase A, and by an acidosis-induced increase in [Ca2+]i and the subsequent activation of a Ca2+/calmodulin-dependent protein kinase, respectively. The possible effect of these changes in phosphorylation on the activity of the Ca2+-ATPase of the cardiac sarcoplasmic reticulum during acidosis is discussed. PMID- 9211816 TI - Direct action of genistein on CFTR. AB - Human cystic fibrosis transmembrane conductance regulator (CFTR) chloride channels were expressed in oocytes from Xenopus laevis after injection of CFTR cRNA and studied with the two-electrode voltage-clamp and the giant patch techniques. The tyrosine kinase inhibitor genistein alone activated a small chloride current in whole oocytes expressing CFTR and substantially increased the chloride current obtained upon stimulation with forskolin and isobutyl methylxanthine (IBMX). In giant excised patches, genistein was unable to open protein-kinase-A-phosphorylated CFTR channels in the absence of ATP, but increased the ATP-induced CFTR channel currents by a factor of 3.8 +/- 1.7. This genistein-mediated potentiation in excised patches is independent of protein phosphatase activity, as it is readily reversible, even after complete inhibition of protein kinase A activity. Involvement of protein tyrosine kinases also seems unlikely, because this effect of genistein is not antagonized by high concentrations of the tyrosine phosphatase inhibitor ortho-vanadate. We, therefore, propose a direct interaction of genistein with CFTR, probably at a nucleotide binding site, which leads to a higher open probability. PMID- 9211817 TI - The effect of brefeldin A on terbutaline-induced sodium absorption in fetal rat distal lung epithelium. AB - We studied the effect of brefeldin A, which inhibits the intracellular trafficking of membrane proteins from the cytosolic pool to the cell surface, on terbutaline (a beta2-specific adrenergic agonist)-induced alterations in ion transport by primary monolayer cultures of fetal rat distal lung epithelium. The amiloride-sensitive short circuit current (Isc) increased 2.5-fold 50 min after application of terbutaline (10 microM) from basolateral side; this response was abolished by pretreatment with brefeldin A (1 microg/ml). Brefeldin A did not suppress the Na+/K+ pump capacity. Single channel patch clamp experiments demonstrated that terbutaline increased the density of amiloride-sensitive Na+ permeable nonselective cation channels on the apical cell membrane and this action was blocked by brefeldin A. These observations suggest that beta2-specific adrenergic agonists promote the trafficking of amiloride sensitive Na+-permeable nonselective cation channels to the apical cell surface. PMID- 9211818 TI - Absence of vacuolar H+-ATPases from rat small intestine brush-border membranes. AB - Rat small intestinal brush-border membrane (BBM) vesicles were solubilized to test for the presence of an intravesicular H+-ATPase. Several detergents that in rat renal BBM vesicles revealed a bafilomycin-sensitive H+-ATPase failed to expose a similar ATPase in small intestinal vesicles. Antibodies against the 31 kDa and 70-kDa H+-ATPase subunits labelled respective antigens in the BBM of renal proximal tubules, but not in the BBM of enterocytes. The results demonstrate that a bafilomycin-sensitive, vacuolar-type H+-ATPase is absent from the BBM of enterocytes and, hence, cannot contribute to H+ secretion. PMID- 9211819 TI - KVLQT channels are inhibited by the K+ channel blocker 293B. AB - Previous data have indicated that the chromanol 293B blocks a cAMP activated K+ conductance in the colonic crypt, a K+ conductance in pig cardiac myocytes and the K+ conductance induced by IsK protein expression in Xenopus oocytes. We have also shown that cAMP-activated cystic fibrosis transmembrane conductance regulator (CFTR) up-regulates, apart from the typical Cl- current, a 293B- inhibitable K+ current. Very recently it has been shown that the IsK protein interacts with KVLQT subunits to produce a K+ channel. These data have prompted us to ask the following questions: Is the 293B-inhibitable current in oocytes expressing CFTR and activated by cAMP caused by an endogenous Xenopus KVLQT (XKVLQT), and is mouse KVLQT (mKVLQT) expressed in oocytes inhibited by 293B? Antisense and sense probes for XKVLQT were coinjected with CFTR cRNA into oocytes. After 3-4 days the oocytes were examined by two electrode voltage clamp. It was found that in control oocytes expressing CFTR and stimulated by isobutylmethylxanthine (IBMX, 1 mmol/l) 293B (10 micromol/l) reduced the conductance (Gm). In oocytes coinjected with the sense probe for XKVLQT and pretreated with IBMX 293B still reduced Gm, whilst the 293B-inhibitable Gm was almost completely absent in oocytes coinjected with XKVLQT antisense. In another series a full length clone for mKVLQT was generated by PCR techniques and the cRNA was injected into oocytes. After several days these oocytes, unlike water injected ones, were found to be strongly hyperpolarized and their Gm was increased significantly. The oocytes were depolarized significantly and their Gm was reduced reversibly by 10 micromol/l 293B. These data indicate that CFTR activation by IBMX indeed co-activates an endogenous oocyte XKVLQT channel and that this channel is inhibited by a new class of channel blockers, of which 293B is the prototype. PMID- 9211820 TI - Role of actin microfilament in osmotic stretch-induced increase of voltage operated calcium channel current in guinea-pig gastric myocytes. AB - Using the whole-cell patch clamp technique, the role of actin microfilament in hyposmotic increase of voltage-operated calcium channel current (IBa) was studied in guinea-pig gastric myocytes. Hyposmotic superfusate (212 mOsm) increased peak IBa amplitude by 32.7 +/- 6.5%; when cytochalasin-D (Cyt-D, 20 microM), an actin cytoskeleton disruptor, was used, an increase of only 9.7 +/- 3.1% was seen. IBa response to osmotic stress was potentiated (45.1 +/- 4.1% increase) by 20 microM phalloidin, an actin microfilament stabilizer. However, colchicine (100 microM), an microtubule cytoskeleton disruptor, had no effect on either IBa or its response to hyposmotic solution. Phalloidin also induced a rightward shift of the I/V relationship of IBa, while Cyt-D itself had no effect. These results suggest that actin cytoskeleton may mediate hyposmotic stretch-induced IBa increase in gastric smooth muscle. PMID- 9211821 TI - Inhibitory effect of phorbol 12,13 dibutyrate on carbachol-activated nonselective cationic current in guinea-pig gastric myocytes. AB - The effect of protein kinase C (PKC) on carbachol (CCh)-activated nonselective cationic current (ICCh) was investigated in guinea-pig gastric myocytes using a PKC activator, phorbol 12, 13 dibutyrate (PDBu). Pretreatment with 1 micro M PDBu suppressed ICCh by 96.5 +/- 2.9% (n = 14) in a reversible manner in nystatin perforated mode. In the presence of 1 microM chelerythrine, a PKC inhibitor, inhibition of ICChby PDBu was not seen. In whole-cell mode, the inhibition of ICCh by PDBu was dependent on intracellular MgATP. In the presence of MgATP in the pipette, PDBu decreased ICCh by 98.8 +/- 1.2% (n = 5) as was observed in nystatin-perforated mode. However, PDBu had little effect on ICCh in the absence of MgATP in the pipette; the extent of inhibition was 12.7 +/- 4.3% (n = 8). PDBu also suppressed the generation of cationic current induced by intracellularly perfused GTP[gammaS]. In the PDBu-pretreated group (n = 9) and PDBu-untreated control group (n = 6), GTP[gammaS]-induced currents were 6.7 +/- 2.4 pA and 236 +/- 23 pA, respectively. These results suggest that PKC modulates ICCh at postreceptor sites via protein phosphorylation. PMID- 9211822 TI - Functional analysis in vivo of the double mutant mouse deficient in both proteolipid protein (PLP) and myelin basic protein (MBP) in the central nervous system. AB - Myelination is an important developmental process of the central (CNS) and peripheral nervous system (PNS). To unravel the functions of the two dominant myelin proteins in the CNS, proteolipid protein (PLP) and myelin basic protein (MBP), we generated and characterized the homozygous double mutant mouse line (plp-/-, mbp-/-), which is viable and fertile. Plasma membrane processes of oligodendrocytes deficient in PLP and MBP, but not in myelin-associated glycoprotein (MAG), spirally wrap large diameter axons, tightly adhering at their extracytosolic surfaces and forming a pseudo-compacted myelin. Neuromotor activity and coordination are considerably improved compared to the shiverer trait. PMID- 9211823 TI - GDNF improves survival and reduces apoptosis in human embryonic dopaminergic neurons in vitro. AB - Dopamine cell death is the primary problem limiting the value of neurotransplantation in human patients with Parkinson's disease. To address this problem, we added glial cell line-derived neurotrophic factor (GDNF) to cultures of embryonic dopaminergic neurons obtained from human and from Bonnet monkey (Macaca radiata) in an effort to reduce apoptotic cell death and improve overall cell survival. Tissue from three human embryos, 7-8 weeks post-conception, and one 9-week post-conception monkey embryo were dissociated and cultured in F-12 media with 5% human placental serum. GDNF (10 ng/ml) in human cultures nearly doubled dopamine neuron survival and reduced the rate of apoptosis from 6% to 3%. In monkey cultures, GDNF also enhanced dopamine neuron survival and reduced the apoptotic rate. We conclude that GDNF improves the survival of primate embryonic dopamine neurons in culture by reducing apoptosis. PMID- 9211824 TI - Prenatal development of the serotonin transporter in mouse brain. AB - The prenatal development of the serotonin transporter was analyzed in mouse brain and spinal cord by autoradiographic localization of [3H]citalopram binding. Transporter expression started at embryonic day (E) 12 in two discontinuous bands in the anterior and posterior brainstem. Labeling extended cranially and caudally, reaching the basal diencephalon at E 13, the septal complex at E 15, and the cerebral cortex at E 16. The caudal extension of the labeling descended at the ventrolateral margin of the spinal cord and reached lumbar levels at E 14. At E 17-E 18, [3H]citalopram binding emerged in the striatum, amygdaloid area, ventrobasal thalamus, paraventricular and periventricular hypothalamic nuclei, and substantia nigra. The overall spatiotemporal expression pattern of the serotonin transporter in the mouse agrees with data on the immunohistochemical localization of serotonin in the rat embryo. These results suggest that serotonergic fibers have the equipment to engage in transmitter reuptake long before synapse formation, and that transporter expression might represent a prerequesite for the developmental functions exerted by serotonin. PMID- 9211825 TI - The columnar region of the ventral nucleus of the lateral lemniscus in the big brown bat (Eptesicus fuscus): synaptic arrangements and structural correlates of feedforward inhibitory function. AB - Neurons of the columnar region of the ventral nucleus of the lateral lemniscus of Eptesicus fuscus respond with high-precision constant-latency responses to sound onsets and possess remarkably broad tuning. To study the synaptic basis for this specialized monaural auditory processing and to elucidate the excitatory or inhibitory nature of the input and output circuitry, we have used classical transmission electron microscopy, and postembedding immunocytochemistry for gamma aminobutyric acid (GABA) and glycine on serial semithin sections. The dominant putatively excitatory perisomatic input is provided by large calyx-like terminals that possess round synaptic vesicles and asymmetric synaptic contacts. Additionally, calyces contact the dendrites of neighboring neurons. Putatively inhibitory small boutons possess pleomorphic or flattened synaptic vesicles and symmetrical contacts and are sparsely distributed on somata and dendrites. Almost all neurons are glycine-immunoreactive. There is a moderate amount of glycine immunoreactive puncta; GABA-immunoreactive puncta are rare. This suggests that (1) there is a fast robust excitatory synaptic input via calyx-like perisomatic endings, (2) calyx-like endings distribute frequency-specific excitatory input across isofrequency sheets by virtue of parallel synapses to somata and adjacent dendrites, and thus, dendritic integration may contribute to the broadening of frequency tuning, (3) the columnar region forms an inhibitory glycinergic feedforward relay in the ascending auditory pathway, a relay that is probably involved in creating filters for time-varying signals. PMID- 9211826 TI - Immunoelectron-microscopic investigation of the subcellular localization of pinopsin in the pineal organ of the chicken. AB - Pinopsin is a photoreceptive molecule cloned from the chicken pineal organ. An antibody highly specific for pinopsin was applied in light- and electron microscopic immunocytochemical studies of the pineal organ of 1 to 2-month-old chickens. Intense immunoreactivity was found in the follicular lumen at the light microscopic level. In addition, small immunoreactive spherical or fibrous structures were diffusely distributed at the parafollicular aspect of the pineal organ. To identify immunoreactive elements precisely, we used pre-embedding immunoelectron microscopy. These studies revealed immunoreactive outer segments of pinealocytes arranged closely side by side in the follicular lumina. The thin initial portion of the outer segment arose from a basal body located in the inner segment. Immunoreactive pear-shaped outer segments occupied small lumina. Follicular lumina displayed immunonegative arrays of whorl-like lamellar membranes. Occasionally, these immunonegative structures were surrounded by immunoreactive concentric lamellar complexes. In the parafollicular pineal parenchyma, long slender cilium-like structures or enlarged cilia and concentric lamellar arrays showed intense immunoreactivity. All immunoreactive structures observed in this study were considered to represent outer segments of pinealocytes of the chicken pineal organ. PMID- 9211827 TI - Laminin immunoreactivity in enteric ganglia of the chick embryo. AB - The localization and time of appearance of laminin in the duodenum of the chick embryo were studied with an anti-laminin polyclonal antibody and immunofluorescence. Laminin immunoreactivity was observed in the basement membranes of the mesothelium, mucosal epithelium, muscle cells and in the adventitia and basal surface of the endothelium in blood vessels. In addition, laminin immunostaining was detected over the contour of myenteric ganglia from embryonic day 7 and inside these ganglia from embryonic day 13. In colocalization experiments, laminin immunoreactivity occurred outside tubulin immunoreactive neuronal cell bodies, thus indicating that it resides in glial cells or in extracellular spaces. In addition connecting strands of the myenteric plexus and intramuscular nerves expressed laminin immunoreactivity. Similar observations were made in the proventriculus, gizzard, ileum and rectum of chick embryos, and in the duodenum and rectum of quail embryos. In the ganglion of Remak, laminin immunofluorescence was detected in the collagenous sheath that surrounds the ganglion and inside the ganglion, where it outlines neuronal cell bodies. Laminin immunoreactivity within the myenteric ganglia during the 3rd week in ovo, appears to be characteristic of the avian species examined, since it was not observed in the rat and mouse intestine at equivalent developmental stages. Immunocytochemical experiments at the electron-microscope level confirmed that structures with laminin or laminin-like immunoreactivity occur both around and inside myenteric ganglia. It is suggested that laminin, or an immunologically similar molecule, may play a role in the development and maturation of avian enteric ganglia. PMID- 9211828 TI - Multiple mechanisms contribute to myenteric plexus ablation induced by benzalkonium chloride in the guinea-pig ileum. AB - Ablation of rat myenteric plexus with benzalkonium chloride has provided a model of intestinal aganglionosis, but the degenerative responses are not well understood. We examined the effects of this detergent on neurons and glia, including expression of c-Myc, c-Jun, JunB, and c-Fos, and on immunocytes in the guinea-pig ileum. Benzalkonium chloride (0.1%) or saline was applied to the serosal surface of distal ileum. Tissues were analyzed 2, 3, or 7 days later and compared with cyclosporine-treated and untreated animals. More than 90% of myenteric neurons were destroyed in ileal segments 3-7 days after benzalkonium chloride treatment. Glia withdrew processes from around neurons after 2 days and were mostly gone after 3 days. Neuronal c-Myc began to disappear while c-Fos, c Jun, and JunB were evident in some neuronal nuclei after 2 or 3 days. After 3 days, widespread apoptosis was evident in the myenteric plexus. Populations of T cells, B cells, and macrophage-like cells in untreated and saline-treated myenteric plexuses were substantially increased 3 and 7 days after benzalkonium chloride treatment. Cyclosporine delayed significant neuronal loss. We conclude that a variety of degenerative mechanisms may be active in this model, including an immune response which may actively contribute to tissue destruction. PMID- 9211829 TI - Development and organization of the retinal dopaminergic system of Ichthyophis kohtaoensis (Amphibia; Gymnophiona). AB - Ichthyophis kohtaoensis, a member of the limbless Gymnophiona, has a specialized subterranean burrowing mode of life and a predominantly olfactory-guided orientation. The only visually guided behavior seems to be negative phototaxis. As these animals possess extremely small eyes (only 540 microm in diameter in adults), functional investigations of single retinal cells by electrophysiological methods have so far failed. Therefore, the content and distribution of retinal transmitters have been investigated as indications for a functioning sense organ in an animal that is supposed to be blind. In this study, the organization and development of the dopaminergic system have been examined in the retinae of embryonic, larval, and adult I. kohtaoensis, by using an antiserum against tyrosine hydroxylase, the rate-limiting enzyme in the catecholamine synthetic pathway, and an antiserum against dopamine itself. Labeled somata are situated in the inner nuclear layer and in the ganglion cell layer. Dopamine positive fibers form a dense diffuse plexus, that covers the whole inner plexiform layer, whereas tyrosine hydroxylase-immunoreactive processes show a tendency to arborize in a stratified manner. Tyrosine-hydroxylase-immunolabeled fibers can occasionally be observed in the optic nerve head of larval stages. During ontogenesis and larval development, the distribution of transmitter expressing cells changes and their number decreases, but no general degeneration of the visual system is detectable. Adult Ichthyophis still have retinal transmitters, indicating that the eyes, although obviously playing a minor role in a subterranean ecological niche, retain all the elements of functioning sense organs. PMID- 9211830 TI - Distribution of neuropeptide Y-like immunoreactivity in the brain of the bichir, Polypterus senegalus, with special regard to the terminal nerve. AB - The distribution of neuropeptide Y (NPY)-like immunoreactivity in the brain of the bichir, Polypterus senegalus, was examined immunohistochemically. NPY-like immunoreactivity was distributed widely in the brain, with the highest density in the diencephalon. NPY-positive perikarya were found in various areas, including the terminal nerve, the pallial zones of the telencephalon, the periventricular preoptic nucleus, the thalamic nucleus, the ventral hypothalamus of the diencephalon, the tegmentum of the mesencephalon, and the area intermedioventralis of the rhombencephalon. In the hypothalamus, NPY-positive liquor-contacting neurons were frequently observed. Immunoreactive neuron-like cells also appeared in the distal lobe of the hypophysis. NPY fibers were densely distributed in the ventral telencephalon, the hypothalamus, and the ventrolateral area of the rhombencephalon. They were also demonstrated in the terminal nerve. In the hypophysis, NPY fibers were dense in the median eminence, but sparse in the neural lobe. Electron-microscopic double immunostaining of the terminal nerve revealed the coexistence of NPY-like antigen with gonadotropin-releasing hormone like and molluscan cardioexcitatory tetrapeptide-like antigens in the same cytoplasmic granules. These results suggest that NPY or a related substance is involved in neuroregulation of various areas of the bichir brain, by mainly acting as a neurotransmitter or neuromodulator. PMID- 9211831 TI - Recruitment of maternal material during assembly of the zygote centrosome in fertilized sea urchin eggs. AB - Spindle poles of sea urchin embryos contain centrosomal material derived from maternal as well as paternal sources. To examine how maternal centrosomal material becomes recruited into spindle poles during the first cell cycle, fertilized sea urchin eggs were fixed and labeled with an anti-centrosomal antibody at sequential timepoints after insemination. Immunolabeling patterns demonstrate that the unfertilized egg contains small foci of immunoreactive material dispersed throughout the cytoplasm. Shortly after insemination, the diffuse foci coalesce to form a dense aggregate close to the sperm nucleus. Subsequently, centrosomal material spreads over the surface of the zygote nucleus and becomes partitioned into two masses during spindle pole formation. The involvement of the cytoskeleton in the translocation and targeting of maternal centrosomal material through the first cell cycle was examined by treating eggs with cytoskeletal disrupting agents, a general kinase inhibitor, and by re inseminating fertilized eggs. These experiments indicate that the initially diffuse centrosomal material is transported centripetally to the sperm nucleus by the sperm aster and the centrosomal material is subsequently sequestered around the zygote nucleus by a microtubule-mediated mechanism. Remarkably, 6 dimethylaminopurine treatment shifted the targeting of maternal centrosomal material from the sperm nucleus to the female pronucleus; upon recovery, some of these zygotes formed spindle poles that flanked only the maternal chromosomes. PMID- 9211832 TI - Coordinated change between complement C1s production and chondrocyte differentiation in vitro. AB - In vitro synthesis of the first component of complement C1s was examined by using hamster epiphyseal chondrocytes (HAC) and human chondrosarcoma cell line HCS-2/8. Hamster and human C1s produced by the cells were quantified by immunoblotting and sandwich enzyme-linked immunosorbent assay (ELISA), respectively. It was possible to measure active and inactive C1s by sandwich ELISA, when we used anti-human C1s monoclonal antibodies, M241 recognizing only active C1s, and M365 and M81 recognizing both active and inactive C1s. Approximately 40% of C1s secreted from HCS-2/8 was found to be activated in the culture medium, whereas C1s from HAC was not. C1s production increased in accordance with chondrocyte differentiation induced by ascorbic acid. In contrast, transforming growth factor-beta1 and basic fibroblast growth factor, which inhibited differentiation, suppressed C1s production. These results confirmed our previous observation showing that C1s synthesis increased with differentiation into hypertrophic chondrocytes in vivo. PMID- 9211833 TI - Immunolocalisation of thrombospondin 1 in human, bovine and rabbit cornea. AB - Light- and electron-microscopic immunohistochemical techniques were used to investigate the distribution of the matricellular protein thrombospondin 1 in normal human, bovine and rabbit cornea. Light-microscopic immunoreactivity for thrombospondin 1 was observed in the epithelial basement membrane, posterior Descemet's membrane and endothelium of human and bovine cornea. The bulk of the stroma, the stromal cells (keratocytes) and the anterior part of Descemet's membrane in human and bovine cornea were devoid of detectable thrombospondin 1 and the protein could not be demonstrated in any of the layers of the rabbit cornea. Electron-microscopic immunogold studies of human and bovine cornea revealed that thrombospondin 1 labelling of corneal endothelial (and basal epithelial) cells included focal deposits at cell membranes. It is postulated that thrombospondin 1 regulates interactions between cells and their basement membrane, and perhaps cell-to-cell interactions, in the normal human and bovine corneal endothelium and basal epithelium. PMID- 9211834 TI - Regeneration of skeletal muscle in two teleost fish: Sparus aurata and Brachydanio rerio. AB - Regeneration of skeletal muscle was studied in the sea bream Sparus aurata, in which extensive post-larval muscle hyperplasia contributes to its large adult size, and in the zebrafish Brachydanio rerio, which shows little post-larval hyperplasia and reaches only a small adult size. Small mechanical lesions of body wall muscle were made under general anaesthesia, and the progress of subsequent regeneration was assessed at various intervals by histology and electron microscopy (for general morphology), by immunostaining for desmin and myosin isoforms (to identify the phenotype of new fibres), and by 5'-bromo-2' deoxyuridine (BrdU) incorporation (to identify proliferating cells). Despite the difference in normal growth-related hyperplasia in these fish, a vigorous regeneration occurred in both species, giving rise to new fibres with an initial myosin composition that differed from that in mature fast-white fibres. However, species differences in myosin expression in these fibres suggest that they may have derived from different myoblast populations. In sea bream, myosin expression in regenerating fibres resembled that seen in new fibres produced in post-larval white muscle, whereas in the zebrafish it resembled that of the primitive monolayer fibres formed during embryonic development. Subsequently, most regenerating fibres gradually transformed into the mature fast-white phenotype in both species. PMID- 9211835 TI - Apoptosis in phagocytotic cells of lymphoid tissues in rainbow trout (Oncorhynchus mykiss) following administration of clodronate liposomes. AB - Macrophage function has been studied in vivo by using liposomes that contain dichloromethylene-bisphosphonate (clodronate liposomes) to deplete macrophage subpopulations. In the present study, the effects of intravenously injected clodronate liposomes on the head kidney and spleen of the rainbow trout (Oncorhynchus mykiss) were studied from 1 h to 7 days after injection. The rapid trapping of liposomes in the splenic ellipsoids was followed by depletion of ellipsoidal sheath macrophages and accumulation of particulate material and IgM in the ellipsoidal wall, findings illustrating the importance of ellipsoidal macrophages in the clearance of filtered substances trapped in the reticular matrix of the ellipsoidal wall. A reduced reactivity for acid phosphatase in the spleen and ultrastructural evidence of cell death in phagocytotic cells of the head kidney and spleen supported the selective effect of clodronate liposomes on macrophages in rainbow trout. Apoptotic bodies were prominent ultrastructural features in tissues collected from clodronate-liposome-treated rainbow trout. The increased presence of apoptotic cells in clodronate-liposome-treated trout compared with trout given liposomes containing phosphate-buffered saline was confirmed by using terminal deoxynucleotidyl-transferase-mediated deoxyuridine triphosphate nick-end-labelling of cells with extensive DNA fragmentation. The characterization of liposome-mediated macrophage depletion in fish provides a useful model for further investigation of piscine macrophage function. PMID- 9211836 TI - Direct evidence for the nature of the binding of mitochondria to microtubules in ovarian nutritive tubes of an hemipteran insect. AB - Interactions between mitochondria and microtubules have been investigated in the nutritive tubes that link the nurse cells to the oocytes in ovarioles of the hemipteran insect, Notonecta. The nutritive tubes comprise large numbers of microtubules, which can be dissected manually from ovarioles. This approach, which retains the intrinsic components of the system, has allowed the in vivo interactions between the microtubules and mitochondria, which are also present in the nutritive tubes, to be studied directly. Static binding occurred between mitochondria and microtubules, and investigations of its nucleotide and salt sensitivities have indicated its microtubule-associated protein (MAP)-dependency. PMID- 9211837 TI - FMRF amide-like-immunoreactive primary sensory neurons in the olfactory system of the terrestrial mollusc, Limax marginatus. AB - The distribution of FMRF amide-like immunoreactivity was investigated in the olfactory organs in the tentacle tip of the terrestrial slug, Limax marginatus. Approximately 0.7% of the neurons in the lobules of the tentacle ganglia demonstrated FMRF amide-like immunoreactivity. Most of the FMRF amide-like immunoreactive somata lay at superficial positions within the lobules, and dendritic processes extended to the outer surface of the sensory epithelium, whereas the axons traveled toward the cerebral ganglion through the ventral part of the tentacle nerve. From their morphological features, FMRF amide-like immunoreactive cells were considered to be primary sensory neurons. PMID- 9211838 TI - Mandible muscle fibers in ants: fast or powerful? AB - Ants use their mandibles for catching prey, cracking seeds, cutting leaves, or for the construction of nests and the tender care of brood. The functional morphology of the mandibles reflect the species' adaptations to particular foraging habits and social life. The versatility and specialization of the mandibles depend directly on the design and physiology of the mandible closer muscles and their component fibers. A comparative video analysis of the closing movements of ant mandibles revealed that the maximal velocity varies considerably among species. The speed is correlated with the morphology of the mandible closer muscle, the largest muscle in ants. It is composed of two morphologically very distinct fiber types: long fibers with short sarcomeres (sarcomere length approximately 2 microM) showing all the structural attributes of fast muscle fibers, and shorter fibers with longer sarcomeres (sarcomere length approximately 5 microM) exhibiting the characteristics of slow and powerful fibers. Ants with fast-moving mandibles have a very high proportion of fast closer fibers, whereas the muscles of ants that cannot perform fast mandible movements have only a few or no fast fibers at all. Fast fibers always attach directly to the solid apodeme, while slow fibers often attach to thin apodeme threads. We suppose that the latter kind of fiber attachment is disadvantageous for fast contracting fibers but helps the ants to make better use of the space in the head capsule. PMID- 9211839 TI - Pattern of bromodeoxyuridine incorporation in the advanced stages of arm regeneration in the feather star Antedon mediterranea. AB - The overall process of arm regeneration in the crinoid Antedon mediterranea is a typical epimorphic process (blastemal regeneration). This can be subdivided into three main phases: a repair phase, an early regenerative phase, and an advanced regenerative phase. The crucial problem of the identification of cell lineages responsible for both repair and regenerative processes has been approached by monitoring cell proliferation during the advanced regenerative phase using light microscopic and ultrastructural immunocytochemical methods to detect the incorporation of the thymidine analogue bromodeoxyuridine (BrdU) into regenerating tissues. Various treatment protocols and BrdU incubation times have been employed and provided information not only on the sources, sites of proliferation, and recruitment times of the new cells, but also on the cell lineage involved and subsequent fate (differentiation and/or migration) of the labelled cells. Our results are consistent with the following conclusions: (1) The arm regeneration process is due to a massive intervention of active proliferating cells identifiable as migratory, morphologically undifferentiated cells (amoebocytes and coelomocytes). (2) The preferential proliferation sites of these cells are the terminal blastema, the coelomic epithelium, and the brachial nerve of both the regenerating arm and the stump, even far from the amputation. (3) The two main cell components contributing to the regenerate have different origins: the blastemal cells and all the cell lineages derived from the amoebocytes; the coelomic cells from the migratory coelomocytes, in their turn derived from proliferation of the coelomic epithelium. (4) The blastemal regeneration of Antedon is due to a combined recruitment of pluripotent elements, implying the intervention of presumptive stem cells (amoebocytes) and the transdifferentiation/dedifferentiation of differentiated elements of the coelomic epithelium. PMID- 9211840 TI - Primary culture of antennal cells of Mamestra brassicae: morphology of cell types and evidence for biosynthesis of pheromone-binding proteins in vitro. AB - A culture technique for the in vitro growth and differentiation of antennal cells of Mamestra brassicae is described. The morphology of the growing cells is described. At least five morphological cell types can be distinguished in the cultures. Neuronal cells (type A) were characterized morphologically and immunocytochemically with anti-HRP staining and could be separated into two subtypes, A1 and A2, on the basis of the size of the cell body. Non-neuronal cells could be divided into four types. Type-B cells consisted of large and flat cells with a veil-like cytoplasm. They usually adhered to each other and formed groups of 20 to 50 cells. Type C were large and flat cells, highly variable in size and shape. They closely resembled insect glial cells in culture. Type-D cells grew thick processes, up to 100 microm long. These elongated processes resembled trichoid hairs, and D cells are most probably trichogen cells. Type E were spindle-shaped cells present in low number in the cultures. A high proportion of similar cells in the hemolymph of pupae suggests that E cells are hemocytes. Pheromone-binding proteins were detected using specific antisera in the medium of 3- to 4-week-old cultures. PMID- 9211842 TI - Orbital steering in the catalytic power of enzymes: small structural changes with large catalytic consequences. AB - Small structural perturbations in the enzyme isocitrate dehydrogenase (IDH) were made in order to evaluate the contribution of precise substrate alignment to the catalytic power of an enzyme. The reaction trajectory of IDH was modified (i) after the adenine moiety of nicotinamide adenine dinucleotide phosphate was changed to hypoxanthine (the 6-amino was changed to 6-hydroxyl), and (ii) by replacing Mg2+, which has six coordinating ligands, with Ca2+, which has eight coordinating ligands. Both changes make large (10(-3) to 10(-5)) changes in the reaction velocity but only small changes in the orientation of the substrates (both distance and angle) as revealed by cryocrystallographic trapping of active IDH complexes. The results provide evidence that orbital overlap produced by optimal orientation of reacting orbitals plays a major quantitative role in the catalytic power of enzymes. PMID- 9211847 TI - Inducible expression and phosphorylation of coactivator BOB.1/OBF.1 in T cells. AB - BOB.1/OBF.1 is a transcriptional coactivator that is constitutively expressed in B cells and interacts with the Oct1 and Oct2 transcription factors. Upon activation of Jurkat T cells and primary murine thymocytes with phorbol esters and ionomycin, BOB.1/OBF.1 expression and transactivation function were induced. BOB.1/OBF.1 was phosphorylated at Ser184 both in vivo and in vitro, and this modification was required for inducible activation. Mutation of Ser184 also diminished transactivation function in B cells, suggesting that the activating phosphorylation that is inducible in T cells is constitutively present in B cells. Thus, BOB.1/OBF.1 is a transcriptional coactivator that is critically regulated by posttranslational modifications to mediate cell type-specific gene expression. PMID- 9211848 TI - Induction of cell migration by matrix metalloprotease-2 cleavage of laminin-5. AB - Structural changes in the extracellular matrix are necessary for cell migration during tissue remodeling and tumor invasion. Specific cleavage of laminin-5 (Ln 5) by matrix metalloprotease-2 (MMP2) was shown to induce migration of breast epithelial cells. MMP2 cleaved the Ln-5 gamma2 subunit at residue 587, exposing a putative cryptic promigratory site on Ln-5 that triggers cell motility. This altered form of Ln-5 is found in tumors and in tissues undergoing remodeling, but not in quiescent tissues. Cleavage of Ln-5 by MMP2 and the resulting activation of the Ln-5 cryptic site may provide new targets for modulation of tumor cell invasion and tissue remodeling. PMID- 9211849 TI - Niemann-Pick C1 disease gene: homology to mediators of cholesterol homeostasis. AB - Niemann-Pick type C (NP-C) disease, a fatal neurovisceral disorder, is characterized by lysosomal accumulation of low density lipoprotein (LDL)-derived cholesterol. By positional cloning methods, a gene (NPC1) with insertion, deletion, and missense mutations has been identified in NP-C patients. Transfection of NP-C fibroblasts with wild-type NPC1 cDNA resulted in correction of their excessive lysosomal storage of LDL cholesterol, thereby defining the critical role of NPC1 in regulation of intracellular cholesterol trafficking. The 1278-amino acid NPC1 protein has sequence similarity to the morphogen receptor PATCHED and the putative sterol-sensing regions of SREBP cleavage-activating protein (SCAP) and 3-hydroxy-3-methyl-glutaryl coenzyme A (HMG-CoA) reductase. PMID- 9211850 TI - Murine model of Niemann-Pick C disease: mutation in a cholesterol homeostasis gene. AB - An integrated human-mouse positional candidate approach was used to identify the gene responsible for the phenotypes observed in a mouse model of Niemann-Pick type C (NP-C) disease. The predicted murine NPC1 protein has sequence homology to the putative transmembrane domains of the Hedgehog signaling molecule Patched, to the cholesterol-sensing regions of 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase and SREBP cleavage-activating protein (SCAP), and to the NPC1 orthologs identified in human, the nematode Caenorhabditis elegans, and the yeast Saccharomyces cerevisiae. The mouse model may provide an important resource for studying the role of NPC1 in cholesterol homeostasis and neurodegeneration and for assessing the efficacy of new drugs for NP-C disease. PMID- 9211851 TI - Abnormal lignin in a loblolly pine mutant. AB - Novel lignin is formed in a mutant loblolly pine (Pinus taeda L.) severely depleted in cinnamyl alcohol dehydrogenase (E.C. 1.1.1.195), which converts coniferaldehyde to coniferyl alcohol, the primary lignin precursor in pines. Dihydroconiferyl alcohol, a monomer not normally associated with the lignin biosynthetic pathway, is the major component of the mutant's lignin, accounting for approximately 30 percent (versus approximately 3 percent in normal pine) of the units. The level of aldehydes, including new 2-methoxybenzaldehydes, is also increased. The mutant pines grew normally indicating that, even within a species, extensive variations in lignin composition need not disrupt the essential functions of lignin. PMID- 9211852 TI - Coding the locations of objects in the dark. AB - The ventral premotor cortex in primates is thought to be involved in sensory motor integration. Many of its neurons respond to visual stimuli in the space near the arms or face. In this study on the ventral premotor cortex of monkeys, an object was presented within the visual receptive fields of individual neurons, then the lights were turned off and the object was silently removed. A subset of the neurons continued to respond in the dark as if the object were still present and visible. Such cells exhibit "object permanence," encoding the presence of an object that is no longer visible. These cells may underlie the ability to reach toward or avoid objects that are no longer directly visible. PMID- 9211853 TI - Pericyte loss and microaneurysm formation in PDGF-B-deficient mice. AB - Platelet-derived growth factor (PDGF)-B-deficient mouse embryos were found to lack microvascular pericytes, which normally form part of the capillary wall, and they developed numerous capillary microaneurysms that ruptured at late gestation. Endothelial cells of the sprouting capillaries in the mutant mice appeared to be unable to attract PDGF-Rbeta-positive pericyte progenitor cells. Pericytes may contribute to the mechanical stability of the capillary wall. Comparisons made between PDGF null mouse phenotypes suggest a general role for PDGFs in the development of myofibroblasts. PMID- 9211854 TI - Activation of heat shock transcription factor 3 by c-Myb in the absence of cellular stress. AB - In vertebrates, the presence of multiple heat shock transcription factors (HSFs) indicates that these factors may be regulated by distinct stress signals. HSF3 was specifically activated in unstressed proliferating cells by direct binding to the c-myb proto-oncogene product (c-Myb). These factors formed a complex through their DNA binding domains that stimulated the nuclear entry and formation of the transcriptionally active trimer of HSF3. Because c-Myb participates in cellular proliferation, this regulatory pathway may provide a link between cellular proliferation and the stress response. PMID- 9211856 TI - Dictyostelium development in the absence of cAMP. AB - Adenosine 3',5'-monophosphate (cAMP) and cAMP-dependent protein kinase (PKA) are regulators of development in many organisms. Dictyostelium uses cAMP as an extracellular chemoattractant and as an intracellular signal for differentiation. Cells that are mutant in adenylyl cyclase do not develop. Moderate expression of the catalytic subunit of PKA in adenylyl cyclase-null cells led to near-normal development without detectable accumulation of cAMP. These results suggest that all intracellular cAMP signaling is effected through PKA and that signals other than extracellular cAMP coordinate morphogenesis in Dictyostelium. PMID- 9211857 TI - Specification of the zebrafish nervous system by nonaxial signals. AB - The organizer of the amphibian gastrula provides the neurectoderm with both neuralizing and posteriorizing (transforming) signals. In zebrafish, transplantations show that a spatially distinct transformer signal emanates from tissues other than the organizer. Cells of the germring (nonaxial mesendoderm) posteriorized forebrain progenitors when grafted nearby, resulting in an ectopic hindbrain-like structure; in contrast, cells of the organizer (axial mesendoderm) caused no posterior transformation. Local application of basic fibroblast growth factor, a candidate transformer in Xenopus, caused malformation but not hindbrain transformation in the forebrain. Thus, the zebrafish gastrula may integrate spatially distinct signals from the organizer and the germring to pattern the neural axis. PMID- 9211858 TI - A reassessment of the low molecular weight phospholipase A2 gene family in mammals. PMID- 9211859 TI - Identification of CCR8, the receptor for the human CC chemokine I-309. AB - The nucleotide sequence for a putative chemokine receptor, termed TER1, ChemR1, or CKR-L1, was recently obtained by a polymerase chain reaction-based cloning technique. It encodes a protein of 355 amino acids that shows 32-45% sequence identity with human chemokine receptors. The gene was localized on human chromosome 3p21-24, the site for the genes for the five known CC chemokine receptors, suggesting that the natural ligand may be a CC chemokine. We have stably expressed this receptor in murine pre-B cells 300-19 and have tested their responsiveness to 20 human chemokines and some other potential agonists. The CC chemokine I-309 was the only agonist that selectively induced intracellular Ca2+ mobilization and chemotaxis in receptor-transfected 300-19 cells. Stromal cell derived factor 1, which binds to murine CXCR4 expressed in parental as well as transfected 300-19 cells, served as positive control in the functional screening. The interaction of I-309 with TER1 was of high affinity as shown by 125I-I-309 binding (Kd of 1.2 nM) and transient [Ca2+]i changes at subnanomolar concentrations of agonist. Migration responses in receptor-transfected 300-19 cells was typically bimodal with maximal activity at 10 nM of I-309. These data demonstrate that TER1 (ChemR1 or CKR-L1) is the receptor for I-309, and we propose to call this receptor CCR8 in agreement with the current nomenclature for chemokine receptors. The expression of CCR8 in blood leukocytes and lymphocytes was analyzed by Northern blot. No transcripts were found in RNA from freshly isolated blood neutrophils, monocytes, cultured macrophages, and phytohemagglutinin-stimulated T lymphocytes, and a faint hybridization signal corresponding to the RNA species of 4 kb was obtained only with RNA from interleukin-2-treated T lymphocytes. CCR8 is unusual for its selectivity for a single chemokine, previously shown only for CXCR1 and CXCR4, which bind interleukin-8 and stromal cell-derived factor 1, respectively. Identification of the receptor for I-309 represents a significant progress in determining the function of I-309 in inflammation and disease. PMID- 9211860 TI - I-FLICE, a novel inhibitor of tumor necrosis factor receptor-1- and CD-95-induced apoptosis. AB - The pivotal discovery that the death proteases caspase 8 (FLICE) and caspase 10 (Mch4/FLICE2) are recruited to the CD-95 and tumor necrosis factor receptor-1 signaling complexes suggested a mechanism used by these cytotoxic receptors to initiate apoptosis. In this report, we describe the cloning and characterization of I-FLICE, a novel inhibitor of tumor necrosis factor receptor-1- and CD-95 induced apoptosis. The overall architecture of I-FLICE is strikingly similar to that of FLICE and Mch4/FLICE2. However, I-FLICE lacks both a catalytic active site and residues that form the substrate binding pocket, in keeping with its dominant negative inhibitory function. I-FLICE is the first example of a catalytically inert caspase that can inhibit apoptosis. PMID- 9211861 TI - The cytoplasmic loop between putative transmembrane segments 6 and 7 in sarcoplasmic reticulum Ca2+-ATPase binds Ca2+ and is functionally important. AB - Limited proteolysis by proteinase K of rabbit SERCA1 Ca2+-ATPase generates a number of fragments which have been identified recently. Here, we have focused on two proteolytic C-terminal fragments, p20C and p19C, starting at Gly-808 and Asp 818, respectively. The longer peptide p20C binds Ca2+, as deduced from changes in migration rate by SDS-polyacrylamide gel electrophoresis performed in the presence of Ca2+ as well as from labeling with 45Ca2+ in overlay experiments. In contrast, the shorter peptide p19C, a proteolysis fragment identical to p20C but for 10 amino acids missing at the N-terminal side, did not bind Ca2+ when submitted to the same experiments. Two cluster mutants of Ca2+-ATPase, D813A/D818A and D813A/D815A/D818A, expressed in the yeast Saccharomyces cerevisiae, were found to have a very low Ca2+-ATPase activity. Region 808-818 is thus essential for both Ca2+ binding and enzyme activity, in agreement with similar results recently reported for the homologous gastric H+, K+-ATPase (Swarts, H. G. P., Klaassen, C. H. W., de Boer, M., Fransen, J. A. M. , and De Pont, J. J. H. H. M. (1996) J. Biol. Chem. 271, 29764-29772). However, the accessibility of proteinase K to the peptidyl link between Leu-807 and Gly-808 clearly shows that the transmembrane segment M6 ends before region 808-818. It is remarkable that critical residues for enzyme activity are located in a cytoplasmic loop starting at Gly-808. PMID- 9211862 TI - The localization of the phosphorylation site of BglG, the response regulator of the Escherichia coli bgl sensory system. AB - BglG, the response regulator of the bgl sensory system, was recently shown to be phosphorylated on a histidine residue. We report here the localization of the phosphorylation site to histidine 208. Localization of the phosphorylated histidine was carried out in two steps. We first engineered BglG derivatives with a specific protease (factor Xa) cleavage site that allowed asymmetric splitting of each prephosphorylated protein to well defined peptides, of which only one was labeled by radioactive phosphate. This allowed the localization of the phosphorylation site to the last 111 residues. Subsequently, we identified the phosphorylated histidine by mutating each of the three histidines located in this region to an arginine and following the ability of the resulting mutants to be in vivo regulated and in vitro phosphorylated by BglF, the bgl system sensor. Histidine 208 was the only histidine which failed both tests. The use of simple techniques to map the phosphorylation site should make this protocol applicable for the localization of phosphorylation sites in other proteins. The bgl system represents a new family of sensory systems. Thus, the mapping reported here is an important step toward the definition of the functional domains involved in the transduction of a signal by the components that constitute systems of this novel family. PMID- 9211863 TI - Phosphorylation and activation of heart 6-phosphofructo-2-kinase by protein kinase B and other protein kinases of the insulin signaling cascades. AB - To understand the insulin-induced activation of 6-phosphofructo-2-kinase (PFK-2) of the bifunctional enzyme PFK-2/fructose-2,6-bisphosphatase in heart, the effect of phosphorylation by protein kinases of the insulin signaling pathways on PFK-2 activity was studied. Purified PFK-2/fructose-2, 6-bisphosphatase from bovine heart is a mixture of two isoforms (Mr 58,000 and 54,000 on SDS-polyacrylamide gels). The Mr 54,000 protein is an alternatively spliced form, lacking phosphorylation sites for protein kinases. Recombinant enzymes corresponding to the Mr 58,000 (BH1) and Mr 54,000 (BH3) forms were expressed and used as substrates for phosphorylation. The recombinant BH1 isoform was phosphorylated by p70 ribosomal S6 kinase (p70(s6k)), mitogen-activated protein kinase-activated protein kinase-1, and protein kinase B (PKB), whereas the recombinant BH3 isoform was a poor substrate for these protein kinases. Treatment with all protein kinases activated PFK-2 in the recombinant BH1 preparation. Phosphorylation of the recombinant BH1 isoform correlated with PFK-2 activation and was reversed by treatment with protein phosphatase 2A. All the protein kinases phosphorylated Ser 466 and Ser-483 in the BH1 isoform, but to different extents: p70(s6k) preferentially phosphorylated Ser-466, whereas mitogen-activated protein kinase activated protein kinase-1 and PKB phosphorylated Ser-466 and Ser-483 to a similar extent. We propose that PKB is part of the insulin signaling cascade for PFK-2 activation in heart. PMID- 9211864 TI - The hemopoietic growth factor, interleukin-3, promotes glucose transport by increasing the specific activity and maintaining the affinity for glucose of plasma membrane glucose transporters. AB - Most mammalian cells rely on an external supply of glucose for survival, proliferation, and function. Glucose enters cells through specific transporter molecules at the plasma membrane by a facilitative process that does not expend energy. Regulation of glucose transport into cells is thought to occur largely through transporter expression at the cell surface, but the extent to which the intrinsic properties of glucose transporters are regulated is at present controversial. Using a bone marrow-derived cell line that responds to the hemopoietic growth factor, interleukin-3 (IL-3), we investigated IL-3 regulation of glucose transport. IL-3 significantly increased 2-deoxyglucose (2-DOG) uptake within 1 h (26 +/- 8.0%, n = 11) with a maximum 73% increase after 6 h. Withdrawal of IL-3 resulted in decreased uptake within 1 h and this continued to decline to 43% of initial uptake by 16 h. To determine whether these changes in 2 DOG uptake were associated with corresponding changes in glucose transporter expression, subtype-specific antisera against Glut-1 and Glut-3 were used. Little change in membrane expression of these transporters was observed prior to 16 h. Fractionation of cell membranes on Nycodenz gradients showed that the majority of each transporter subtype was associated with the plasma membrane (63-93%) and that transporter distribution did not change markedly in response to addition or withdrawal of IL-3. These results demonstrate that IL-3 regulates glucose uptake by modulating the intrinsic transporting ability of glucose transporters. Decreased transporter affinity for 2-DOG and 3-O-methylglucose was observed following IL-3 withdrawal. Similar affinity changes were observed with 2-DOG following exposure of IL-3-stimulated cells to the protein kinase inhibitors, genistein and staurosporine. In contrast, the tyrosine phosphatase inhibitor, vanadate, acted like IL-3 to increase transporter affinity for glucose. Together these results demonstrate that IL-3 acts to maintain the intrinsic transport properties of glucose transporters without markedly affecting their expression or translocation. PMID- 9211865 TI - Defining the topology of integrin alpha5beta1-fibronectin interactions using inhibitory anti-alpha5 and anti-beta1 monoclonal antibodies. Evidence that the synergy sequence of fibronectin is recognized by the amino-terminal repeats of the alpha5 subunit. AB - The high affinity interaction of integrin alpha5beta1 with the central cell binding domain (CCBD) of fibronectin requires both the Arg-Gly-Asp (RGD) sequence (in the 10th type III repeat) and a second site (in the adjacent 9th type III repeat) which synergizes with RGD. We have attempted to map the fibronectin binding interface on alpha5beta1 using monoclonal antibodies (mAbs) that inhibit ligand recognition. The binding of two anti-alpha5 mAbs (P1D6 and JBS5) to alpha5beta1 was strongly inhibited by a tryptic CCBD fragment of fibronectin (containing both synergy sequence and RGD) but not by GRGDS peptide. Using recombinant wild type and mutated fragments of the CCBD, we show that the synergy region of the 9th type III repeat is involved in blocking the binding of P1D6 and JBS5 to alpha5beta1. In contrast, binding of the anti-beta1 mAb P4C10 to alpha5beta1 was inhibited to a similar extent by GRGDS peptide, the tryptic CCBD fragment, or recombinant proteins lacking the synergy region, indicating that the RGD sequence is involved in blocking P4C10 binding. P1D6 inhibited the interaction of a wild type CCBD fragment with alpha5beta1 but had no effect on the binding of a mutant fragment that lacked the synergy region. The epitopes of P1D6 and JBS5 mapped to the NH2-terminal repeats of the alpha5 subunit. Our results indicate that the synergy region is recognized primarily by the alpha5 subunit (in particular by its NH2-terminal repeats) but that the beta1 subunit plays the major role in binding of the RGD sequence. These findings provide new insights into the mechanisms, specificity, and topology of integrin-ligand interactions. PMID- 9211866 TI - Refinement and comparisons of the crystal structures of pig cytosolic aspartate aminotransferase and its complex with 2-methylaspartate. AB - Two high resolution crystal structures of cytosolic aspartate aminotransferase from pig heart provide additional insights into the stereochemical mechanism for ligand-induced conformational changes in this enzyme. Structures of the homodimeric native structure and its complex with the substrate analog 2 methylaspartate have been refined, respectively, with 1.74-A x-ray diffraction data to an R value of 0.170, and with 1.6-A data to an R value of 0.173. In the presence of 2-methylaspartate, one of the subunits (subunit 1) shows a ligand induced conformational change that involves a large movement of the small domain (residues 12-49 and 327-412) to produce a "closed" conformation. No such transition is observed in the other subunit (subunit 2), because crystal lattice contacts lock it in an "open" conformation like that adopted by subunit 1 in the absence of substrate. By comparing the open and closed forms of cAspAT, we propose a stereochemical mechanism for the open-to-closed transition that involves the electrostatic neutralization of two active site arginine residues by the negative charges of the incoming substrate, a large change in the backbone (phi,psi) conformational angles of two key glycine residues, and the entropy driven burial of a stretch of hydrophobic residues on the N-terminal helix. The calculated free energy for the burial of this "hydrophobic plug" appears to be sufficient to serve as the driving force for domain closure. PMID- 9211867 TI - Induction of tyrosine phosphorylation and Na+/H+ exchanger activation during shrinkage of human neutrophils. AB - The ubiquitous isoform of the Na+/H+ exchanger (NHE1) is essential for the regulation of cellular volume. The underlying molecular mechanism, which is poorly understood, was studied in human polymorphonuclear leukocytes (PMN). Suspension of PMN in hypertonic media induced rapid cellular shrinkage and activation of NHE1, which is measurable as a cytosolic alkalinization. Concomitantly, hypertonic stress also induced extensive tyrosine phosphorylation of several proteins. Pretreatment of PMN with genistein, a tyrosine kinase inhibitor, prevented not only the tyrosine phosphorylation in response to a hypertonic shock but also the activation of NHE1. The signal elicited by hyperosmolarity that induces activation of tyrosine kinases and NHE1 was investigated. Methods were devised to change medium osmolarity without altering cell volume and vice versa. Increasing medium and intracellular osmolarity in normovolemic cells failed to activate tyrosine kinases or NHE1. However, shrinkage of cells under iso-osmotic conditions stimulated both tyrosine phosphorylation and NHE1 activity. These findings imply that cells detect alterations in cell size but not changes in osmolarity or ionic strength. The identity of the proteins that were tyrosine-phosphorylated in response to cell shrinkage was also investigated. Unexpectedly, the mitogen-activated protein kinases SAPK, p38, erk1, and erk2 were not detectably phosphorylated or activated. In contrast, the tyrosine kinases p59(fgr) and p56/59(hck) were phosphorylated and activated upon hypertonic challenge. We propose that cells respond to alterations in cell size, but not to changes in osmolarity, with increased tyrosine phosphorylation, which in turn leads to the activation of NHE1. The resulting changes in ion content and cytosolic pH contribute to the restoration of cell volume in shrunken cells. PMID- 9211869 TI - Ablation of Goalpha overrides G1 restriction point control through Ras/ERK/cyclin D1-CDK activities. AB - We have generated stable IIC9 cell lines, Goa1 and Goa2, that overexpress full length antisense Goalpha RNA. As shown previously, expression of antisense Goalpha RNA ablated the alpha subunit of the heterotrimeric G protein, Go, resulting in growth in the absence of mitogen. To better understand this change in IIC9 phenotype, we have characterized the signaling pathway and cell cycle events previously shown to be important in control of IIC9 G1/S phase progression. In this paper we clearly demonstrate that ablation of Goalpha results in growth, constitutively active Ras/ERK, elevated expression of cyclin D1, and constitutively active cyclin D1-CDK complexes, all in the absence of mitogen. Furthermore, these characteristics were abolished by the transient overexpression of the transducin heterotrimeric G protein alpha subunit strongly suggesting the transformation of Goalpha-ablated cells involves Gobetagamma subunits. This is the first study to implicate a heterotrimeric G protein in tumor suppression. PMID- 9211868 TI - Ablation of Go alpha-subunit results in a transformed phenotype and constitutively active phosphatidylcholine-specific phospholipase C. AB - Modulation of the components involved in mitogenic signaling cascades is critical to the regulation of cell growth. GTP-binding proteins and the stimulation of phosphatidylcholine (PC) hydrolysis have been shown to play major roles in these cascades. One of the enzymes involved in PC hydrolysis, a PC-specific phospholipase C (PC-PLC) has received relatively little attention. In this paper we examined the role of a particular heterotrimeric GTP-binding protein, Go, in the regulation of cell growth and PC-PLC-mediated hydrolysis of PC in IIC9 fibroblasts. The Go alpha-subunit was ablated in IIC9 cells by stable expression of antisense RNA. These stably transfected cells acquired a transformed phenotype as indicated by: (a) the formation of multiple foci in monolayer cultures, (b) the acquisition of anchorage-independent growth in soft agar; and (c) an increased level of thymidine incorporation in the absence of added mitogens. These data implicate Goalpha as a novel tumor suppressor. Interestingly, PC-PLC activity was constitutively active in the Goalpha-ablated cells as evidenced by the chronically elevated levels of diacylglycerol and phosphorylcholine in the absence of growth factors. In contrast, basal activities of PC-phospholipase D, phospholipase A2, or phosphoinositol-PLC were not affected. These data demonstrate, for the first time, a role for Go in regulating cell growth and provide definitive evidence for the existence of a PC-PLC in eukaryotic cells. The data further indicate that a subunit of Go, is involved in regulating this enzyme. PMID- 9211870 TI - Protein kinase A activation of the surfactant protein B gene is mediated by phosphorylation of thyroid transcription factor 1. AB - Thyroid transcription factor 1 (TTF-1) is a homeodomain-containing nuclear transcription factor expressed in epithelial cells of the lung and thyroid. TTF-1 binds to and activates the transcription of genes expressed selectively in the respiratory epithelium including pulmonary surfactant A, B, C and Clara cell secretory protein. Transfection with a plasmid encoding the cyclic AMP-dependent protein kinase (protein kinase A; PKA) catalytic subunit, Cat-beta, stimulated the phosphorylation of a TTF-1-flag fusion protein 6-7-fold in H441 pulmonary adenocarcinoma cells. Recombinant TTF-1 was phosphorylated by purified PKA catalytic subunit in the presence of [gamma-32P]ATP. PKA catalytic subunit family members, Cat-alpha and Cat-beta, markedly enhanced the transcriptional activation of surfactant B gene promoters by TTF-1 in vitro. Peptide mapping was used to identify a PKA phosphorylation site at the NH2 terminus of TTF-1. A 17-amino acid synthetic peptide comprising this site completely inhibited the PKA-dependent phosphorylation of TTF-1 in vitro. A substitution mutation of TTF-1 (Thr9 two head right arrow Ala) abolished phosphorylation by PKA and reduced transactivation of the surfactant B gene promoter. Transfection with a plasmid encoding the cAMP regulatory element binding factor inhibited transcriptional activity of the surfactant protein B gene promoter. Phosphorylation of TTF-1 mediates PKA-dependent activation of surfactant protein B gene transcription. PMID- 9211871 TI - Dynamics of the U1 small nuclear ribonucleoprotein during yeast spliceosome assembly. AB - U1 small nuclear ribonucleoprotein (snRNP) may function during several steps of spliceosome assembly. Most spliceosome assembly assays, however, fail to detect the U1 snRNP. Here, I used a new native gel electrophoretic assay to find the yeast U1 snRNP in three pre-splicing complexes (delta, beta1, alpha2) formed in vitro. The order of complex formation is deduced to be delta --> beta1 --> alpha2 --> alpha1 --> beta2, the active spliceosome. The delta complex is formed when U1 snRNP binds to pre-mRNA in the absence of ATP. There are two forms of delta: a major one, deltaun, unstable to competitor RNA; and a minor one, deltacommit, committed to the splicing pathway. The other complexes are formed in the presence of ATP and contain the following snRNPs: beta1, the pre-spliceosome, has both U1 and U2; alpha2 has all five, however, U1 is reduced compared with the others; and alpha1 and beta2 have U2, U5, and U6. Prior work by others suggests that U1 is "handing off" the 5' splice site region to the U5 and U6 snRNPs before splicing begins. The reduced levels of U1 snRNP in the alpha2 complex suggests that the handoff occurs during formation of this complex. PMID- 9211872 TI - Cloning of the human prolyl 4-hydroxylase alpha subunit isoform alpha(II) and characterization of the type II enzyme tetramer. The alpha(I) and alpha(II) subunits do not form a mixed alpha(I)alpha(II)beta2 tetramer. AB - Prolyl 4-hydroxylase (proline hydroxylase, EC 1.14.11.2) catalyzes the formation of 4-hydroxyproline in collagens. The vertebrate enzyme is an alpha2beta2 tetramer, the beta subunit of which is identical to protein disulfide-isomerase (PDI, EC 5.3.4.1). We report here on cloning of the recently discovered alpha(II) subunit from human sources. The mRNA for the alpha(II) subunit was found to be expressed in a variety of human tissues, and the presence of the corresponding polypeptide and the (alpha(II))2beta2 tetramer was demonstrated in cultured human WI-38 and HT-1080 cells. The type II tetramer was found to represent about 30% of the total prolyl 4-hydroxylase in these cells and about 5-15% in various chick embryo tissues. The results of coexpression in insect cells argued strongly against the formation of a mixed alpha(I)alpha(II)beta2 tetramer. PDI/beta polypeptide containing a histidine tag in its N terminus was found to form prolyl 4-hydroxylase tetramers as readily as the wild-type PDI/beta polypeptide, and histidine-tagged forms of prolyl 4-hydroxylase appear to offer an excellent source for a simple large scale purification of the recombinant enzyme. The properties of the purified human type II enzyme were very similar to those of the type I enzyme, but the Ki of the former for poly(L-proline) was about 200-1000 times that of the latter. In agreement with this, a minor difference, about 3-6 fold, was found between the two enzymes in the Km values for three peptide substrates. The existence of two forms of prolyl 4-hydroxylase in human cells raises the possibility that mutations in one enzyme form may not be lethal despite the central role of this enzyme in the synthesis of all collagens. PMID- 9211873 TI - The ferrous-dioxy complex of neuronal nitric oxide synthase. Divergent effects of L-arginine and tetrahydrobiopterin on its stability. AB - Nitric oxide synthases (NOS) are hemeproteins that catalyze oxidation of L arginine to nitric oxide (NO) and citrulline. The NOS heme iron is expected to participate in oxygen activation during catalysis, but its interactions with O2 are not characterized. We utilized the heme-containing oxygenase domain of neuronal NOS (nNOSoxy) and stopped-flow methods to study formation and autooxidative decomposition of the nNOSoxy oxygenated complex at 10 degrees C. Mixing ferrous nNOSoxy with air-saturated buffer generated a transient species with absorption maxima at 427 and approximately 560 nm. This species decayed within 1 s to form ferric nNOSoxy. Its formation was first order with respect to O2, monophasic, and gave rate constants for kon = 9 x 10(5) M-1 s-1 and koff = 108 s-1 for an L-arginine- and tetrahydrobiopterin (H4B)-saturated nNOSoxy. Omission of L-arginine and/or H4B did not greatly effect O2 binding and dissociation rates. Decomposition of the oxygenated intermediate was independent of O2 concentration and was either biphasic or monophasic depending on sample conditions. L-Arginine stabilized the oxygenated intermediate (decay rate = 0.14 s-1), while H4B accelerated its decay by a factor of 70 irrespective of L arginine. The spectral and kinetic properties of the intermediate identify it as the FeIIO2 complex of nNOSoxy. Destabilization of a metallo-oxy species by H4B is unprecedented and may be important regarding the role of this cofactor in NO synthesis. PMID- 9211874 TI - Diacylglycerol and phosphatidate generated by phospholipases C and D, respectively, have distinct fatty acid compositions and functions. Phospholipase D-derived diacylglycerol does not activate protein kinase C in porcine aortic endothelial cells. AB - Stimulation of cells with certain agonists often activates both phospholipases C and D. These generate diacylglycerol and phosphatidate, respectively, although the two lipids are also apparently interconvertable through the actions of phosphatidate phosphohydrolase and diacylglycerol kinase. Diacylglycerol activates protein kinase C while one role for phosphatidate is the activation of actin stress fiber formation. Therefore, if the two lipids are interconvertable, it is theoretically possible that an uncontrolled signaling loop could arise. To address this issue structural analysis of diacylglycerol, phosphatidate, and phosphatidylbutanol (formed in the presence of butan-1-ol) from both Swiss 3T3 and porcine aortic endothelial cells was performed. This demonstrated that phospholipase C activation generates primarily polyunsaturated species while phospholipase D activation generates saturated/monounsaturated species. In the endothelial cells, where phospholipase D was activated by lysophosphatidic acid independently of phospholipase C, there was no activation of protein kinase C. Thus we propose that only polyunsaturated diacylglycerols and saturated/monounsaturated phosphatidates function as intracellular messengers and that their interconversion products are inactive. PMID- 9211875 TI - Glycoprotein 46 mRNA abundance is post-transcriptionally regulated during development of Leishmania chagasi promastigotes to an infectious form. AB - GP46 is an abundant glycoprotein of 46 kDa on the surface of the promastigote form of most Leishmania species. We show that the steady state level of GP46 mRNA increases >>30-fold as Leishmania chagasi promastigotes develop in vitro from a less infectious form during logarithmic growth to a highly infectious form in the stationary phase of cultivation. Nuclear run-on experiments demonstrate that this increase in GP46 mRNA abundance is regulated post-transcriptionally. Plasmids containing the 3'-untranslated regions (UTRs) and downstream intergenic regions (IRs) of two different GP46 genes fused immediately downstream of the beta galactosidase coding region were transfected into L. chagasi, and beta galactosidase activity and mRNA levels were examined. The presence of the 3'-UTR + IR of one GP46 gene (gp46A) resulted in a steady increase in beta-galactosidase activity and mRNA level as the transfected promastigotes developed from logarithmic to stationary phase. This differential effect parallels that of the 3'-UTRs + IRs of a family of genes for an unrelated Leishmania surface glycoprotein, GP63. Thus, post-transcriptional regulation of the genes for two different surface glycoproteins of Leishmania occurs via a similar mechanism. PMID- 9211876 TI - Isoprenylated human brain type I inositol 1,4,5-trisphosphate 5-phosphatase controls Ca2+ oscillations induced by ATP in Chinese hamster ovary cells. AB - D-myo-Inositol 1,4,5-trisphosphate (InsP3) 5-phosphatase and 3-kinase are thought to be critical regulatory enzymes in the control of InsP3 and Ca2+ signaling. In brain and many other cells, type I InsP3 5-phosphatase is the major phosphatase that dephosphorylates InsP3 and D-myo-inositol 1,3,4,5-tetrakisphosphate. The type I 5-phosphatase appears to be associated with the particulate fraction of cell homogenates. Molecular cloning of the human brain enzyme identifies a C terminal farnesylation site CVVQ. Post-translational modification of this enzyme promotes membrane interactions and changes in specific activity. We have now compared the cytosolic Ca2+ ([Ca2+]i) responses induced by ATP, thapsigargin, and ionomycin in Chinese hamster ovary (CHO-K1) cells transfected with the intact InsP3 5-phosphatase and with a mutant in which the C-terminal cysteine cannot be farnesylated. [Ca2+]i was also measured in cells transfected with an InsP3 3 kinase construct encoding the A isoform. The Ca2+ oscillations detected in the presence of 1 microM ATP in control cells were totally lost in 87.5% of intact (farnesylated) InsP3 5-phosphatase-transfected cells, while such a loss occurred in only 1.1% of the mutant InsP3 5-phosphatase-transfected cells. All cells overexpressing the InsP3 3-kinase also responded with an oscillatory pattern. However, in contrast to control cells, the [Ca2+]i returned to base-line levels in between a couple of oscillations. The [Ca2+]i responses to thapsigargin and ionomycin were identical for all cells. The four cell clones compared in this study also behaved similarly with respect to capacitative Ca2+ entry. In permeabilized cells, no differences in extent of InsP3-induced Ca2+ release nor in the threshold for InsP3 action were observed among the four clones and no differences in the expression levels of the various InsP3 receptor isoforms could be shown between the clones. Our data support the contention that the ATP-induced increase in InsP3 concentration in transfected CHO-K1 cells is essentially restricted to the site of its production near the plasma membrane, where it can be metabolized by the type I InsP3 5-phosphatase. This enzyme directly controls the [Ca2+]i response and the Ca2+ oscillations in intact cells. PMID- 9211877 TI - ELO2 and ELO3, homologues of the Saccharomyces cerevisiae ELO1 gene, function in fatty acid elongation and are required for sphingolipid formation. AB - ELO2 and ELO3 were identified from the Saccharomyces cerevisiae genome data base as homologues of ELO1, a gene involved in the elongation of the fatty acid 14:0 to 16:0. Mutations in these genes have previously been shown to produce pleiotropic effects involving a number of membrane functions. The simultaneous disruption of ELO2 and ELO3 has also been shown to produce synthetic lethality, indicating that they have related and/or overlapping functions. Gas chromatography and gas chromatography/mass spectroscopy analyses reveal that null mutations of ELO2 and ELO3 produce defects in the formation of very long chain fatty acids. Analysis of the null mutants indicates that these genes encode components of the membrane-bound fatty acid elongation systems that produce the 26-carbon very long chain fatty acids that are precursors for ceramide and sphingolipids. Elo2p appears to be involved in the elongation of fatty acids up to 24 carbons. It appears to have the highest affinity for substrates with chain lengths less than 22 carbons. Elo3p apparently has a broader substrate specificity and is essential for the conversion of 24-carbon acids to 26-carbon species. Disruption of either gene reduces cellular sphingolipid levels and results in the accumulation of the long chain base, phytosphingosine. Null mutations in ELO3 result in accumulation of labeled precursors into inositol phosphoceramide, with little labeling in the more complex mannosylated sphingolipids, whereas disruption of ELO2 results in reduced levels of all sphingolipids. PMID- 9211878 TI - Quaternary structure regulates hemin dissociation from human hemoglobin. AB - Rate constants for hemin dissociation from the alpha and beta subunits of native and recombinant human hemoglobins were measured as a function of protein concentration at pH 7.0, 37 degrees C, using H64Y/V68F apomyoglobin as a hemin acceptor reagent. Hemin dissociation rates were also measured for native isolated alpha and beta chains and for recombinant hemoglobin tetramers stabilized by alpha subunit fusion. The rate constant for hemin dissociation from beta subunits in native hemoglobin increases from 1.5 h-1 in tetramers at high protein concentration to 15 h-1 in dimers at low concentrations. The rate of hemin dissociation from alpha subunits in native hemoglobin is significantly smaller (0.3-0.6 h-1) and shows little dependence on protein concentration. Recombinant hemoglobins containing a fused di-alpha subunit remain tetrameric under all concentrations and show rates of hemin loss similar to those observed for wild type and native hemoglobin at high protein concentration. Rates of hemin dissociation from monomeric alpha and beta chains are much greater, 12 and 40 h 1, respectively, at pH 7, 37 degrees C. Aggregation of monomers to form alpha1beta1 dimers greatly stabilizes bound hemin in alpha chains, decreasing its rate of hemin loss approximately 20-fold. In contrast, dimer formation has little stabilizing effect on hemin binding to beta subunits. A significant reduction in the rate of hemin loss from beta subunits does occur after formation of the alpha1beta2 interface in tetrameric hemoglobin. These results suggest that native human hemoglobin may have evolved to lose heme rapidly after red cell lysis, allowing the prosthetic group to be removed by serum albumin and apohemopexin. PMID- 9211879 TI - Evidence for a unique long chain acyl-CoA ester binding site on the ATP-regulated potassium channel in mouse pancreatic beta cells. AB - The mechanism by which long chain acyl-CoA (LC-CoA) esters affect the ATP regulated potassium channel (KATP channel) was studied in inside-out patches isolated from mouse pancreatic beta cells. Addition of LC-CoA esters dramatically increased KATP channel activity. The stimulatory effect of the esters could be explained by the induction of a prolonged open state of the channel and did not involve alterations in single channel unitary conductance. Under control conditions, absence of adenine nucleotides, the distribution of KATP channel open time could be described by a single exponential, with a time constant of about 25 ms. Exposing the same patch to LC-CoA esters resulted in the appearance of an additional component with a time constant of >>150 ms, indicating a conformational change of the channel protein. LC-CoA esters were also able to potently activate channel activity at different ratios of ATP/ADP. Simultaneous additions of MgADP and LC-CoA esters resulted in a supra-additive effect on channel mean open time, characterized by openings of very long duration. Following modification of the KATP channel by a short exposure of the patch to the protease trypsin, the stimulatory effect of ADP on channel activity was lost while activation by LC-CoA esters still persisted. This indicates that LC-CoA esters and MgADP do not bind to the same site. We conclude that LC-CoA esters may play an important role in the physiological regulation of the KATP channel in the pancreatic beta cell by binding to a unique site and thereby inducing repolarization of the beta cell-membrane potential. PMID- 9211881 TI - An antibody reactive with domain 4 of the platelet-derived growth factor beta receptor allows BB binding while inhibiting proliferation by impairing receptor dimerization. AB - A panel of murine monoclonal antibodies was generated against the extracellular domain of the human platelet-derived growth factor (PDGF) beta receptor (PDGFRbeta). These antibodies were assayed for both the ability to inhibit binding of PDGF BB to PDGFRbeta+ cells as well as the capacity to inhibit PDGF BB mediated mitogenesis. As expected, all antibodies that could prevent PDGF BB binding also inhibited mitogenesis. However one antibody (M4TS.11), with no detectable ability to inhibit PDGF BB binding, was a potent inhibitor of proliferation induced by PDGF BB. Further characterization indicated that M4TS.11 impaired PDGFRbeta dimerization, revealing the mechanism by which it prevented PDGF BB-mediated mitogenesis. Using domain deletion mutants of the extracellular portion of PDGFRbeta, the determinant recognized by this antibody was localized to the fourth extracellular domain of PDGFRbeta, indicating that this domain, which is not involved in ligand binding, actively participates in receptor dimerization and signal transduction. The M4TS.11 antibody could also inhibit PDGF BB-mediated proliferation of responsive cells from both the baboon and the rabbit, indicating the determinant recognized by the antibody is not limited to humans and making it possible to use this antibody to evaluate the therapeutic benefit of interfering with PDGF in animal models of human disease. PMID- 9211880 TI - Thiol activation of endopeptidase EC 3.4.24.15. A novel mechanism for the regulation of catalytic activity. AB - Endopeptidase EC 3.4.24.15 (EP24.15) is a thermolysin-like metalloendopeptidase involved in the regulated metabolism of a number of neuropeptides. Unlike other thermolysin-like peptidases EP24.15 displays a unique thiol activation, a mechanism that is not clearly understood. In this study we show that both recombinant and tissue-derived EP24.15 are activated up to 8-fold by low concentrations (0.1 mM) of dithiothreitol. Additionally, under non-reducing conditions, recombinant and native EP24.15 forms multimers that can be returned to the monomeric form by reduction. We have also shown that competitive inhibitor binding occurs only to the monomeric form, which indicates that catalytic site access is restricted in the multimeric forms. Through systematic site-directed mutagenesis we have identified that cysteine residues 246, 253, and possibly 248 are involved in the formation of these multimers. Furthermore, both a double mutant (C246S/C253S) and a triple mutant (C246S/C248S/C253S) are fully active in the absence of reducing agents, as measured by both inhibitor binding and hydrolysis. The formation and disruption of disulfide bonds involving these cysteine residues may be a mechanism by which EP24.15 activity is regulated through changes in intra- and extracellular redox potential. PMID- 9211882 TI - Identification of four amino acids in the gastrin-releasing peptide receptor that are required for high affinity agonist binding. AB - The bombesin family of G-protein-coupled receptors includes the gastrin-releasing peptide receptor (GRP-R), the neuromedin B receptor (NMB-R), bombesin receptor subtype 3 (BRS-3), and bombesin receptor subtype 4 (bb4). All species homologues of GRP-R, NMB-R, and bb4 bind bombesin with dissociation constants in the nanomolar range; by comparison, human BRS-3 binds bombesin at much lower affinity (Kd >> 1 microM). We used this difference to help identify candidate residues that were potentially critical for forming the bombesin binding pocket. We reasoned that amino acids essential for bombesin binding would be conserved among all homologues of bb4, NMB-R, and GRP-R; conversely, at least one of these amino acids would not be conserved among homologues of BRS-3. Amino acid sequence alignment revealed nine residues that fit this model. We replaced each of these amino acids in mouse GRP-R with the homologous amino acid in human BRS-3. Four substitutions resulted in a significant decrease in bombesin affinity (R288H, Q121R, P199S, and A308S). The analogous mutations in BRS-3 (R127Q, H294R, S205P, and S315A) together resulted in a receptor with a 100-fold increase in bombesin and GRP affinities relative to wild-type BRS-3. From this, we propose a preliminary map of some of the amino acids comprising the agonist binding pocket. PMID- 9211883 TI - N-terminal hydrophobic sorting signals of preproteins confer mitochondrial hsp70 independence for import into mitochondria. AB - The requirement of mitochondrial hsp70 (mt-hsp70) for the import of a series of preproteins containing hydrophobic sorting signals into isolated yeast mitochondria was investigated. Here we demonstrate that the presence of such a sorting signal in proximity to the N-terminal matrix-targeting sequence of a preprotein can secure a translocating polypeptide chain in the import channel in a manner that does not require mt-hsp70 activity. Trapping the translocating chain in this fashion leads to efficient processing by the mitochondrial processing peptidase and to complete translocation across the outer mitochondrial membrane into the intermembrane space. These mt-hsp70-independent effects appear to be exerted at the level of the inner membrane through an interaction of the hydrophobic core of the sorting signal with component(s) of the translocase of the inner membrane. Hydrophobic sorting signals of inner membrane proteins inserted into the membrane from the matrix, as well as those of intermembrane space proteins, are capable of causing this mt-hsp70-independent stabilization, demonstrating that this phenomenon is not unique to those preproteins normally sorted to the intermembrane space. PMID- 9211884 TI - Distinct roles for MAX protein isoforms in proliferation and apoptosis. AB - MAX is a basic helix-loop-helix-leucine zipper protein that plays a central role in the transcriptional control of Myc oncoproteins. MYC-MAX heterodimers stimulate transcription, whereas MAX homodimers, or heterodimers between MAX and members of the MAD family of basic helix-loop-helix-leucine zipper proteins, repress transcription. Max exists in two major isomeric forms, MAX(L) and MAX(S), which differ from one another only by a 9-amino acid insertion/deletion. We show here that MAX(L) is much more effective at homodimeric DNA binding than MAX(S). In NIH3T3 cells, MAX(L) was able to repress a c-Myc-responsive reporter gene whereas MAX(S) either stimulated the reporter gene or had little effect on its expression. In comparison to control cell lines or those stably over-expressing MAX(S), MAX(L)-over-expressing cell lines showed reduced expression of transiently expressed or endogenous c-Myc responsive genes, grew more slowly, possessed a higher growth factor requirement, and showed accelerated apoptosis following growth factor deprivation. Differential effects on growth and apoptosis represent two previously unrecognized properties of MAX proteins. These can at least partly be explained by the differences in their DNA binding abilities and their effects on target gene expression. PMID- 9211885 TI - Erythromycin resistance peptides selected from random peptide libraries. AB - Translation of a 5-codon mini-gene encoded in Escherichia coli 23 S rRNA was previously shown to render cells resistant to erythromycin (Tenson, T., DeBlasio, A., and Mankin, A. S. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 5641-5646). Erythromycin resistance was mediated by a specific interaction of the 23 S rRNA encoded pentapeptide with the ribosome. In the present study, peptides conferring erythromycin resistance were selected from in vivo expressed random peptide libraries to study structural features important for peptide activity. Screening of a 21-codon mini-gene library (the general structure ATG (NNN)20 TAA) demonstrated that only short peptides (3-6 amino acids long) conferred erythromycin resistance. Sequence comparison of erythromycin resistance peptides isolated from the 5-codon library (ATG (NNN)4 TAA) revealed a strong preference for leucine or isoleucine as a third amino acid and a hydrophobic amino acid at the C terminus of the peptide. When tested against other antibiotics, erythromycin resistance peptides rendered cells resistant to other macrolides, oleandomycin and spiramycin, but not to chloramphenicol or clindamycin. Defining the consensus amino acid sequence of erythromycin resistance peptides provided insights into a possible mode of peptide action and the nature of the peptide binding site on the ribosome. PMID- 9211887 TI - Cell-specific expression of the glucose-dependent insulinotropic polypeptide gene in a mouse neuroendocrine tumor cell line. AB - Glucose-dependent insulinotropic polypeptide (GIP) is a 42-amino acid gastrointestinal regulatory peptide that, in the presence of glucose, stimulates insulin secretion. GIP is expressed in K cells of the small intestine and in cells of the submandibular salivary gland. Using a rat GIP cDNA as a specific probe, we screened a number of established cell lines for the expression of GIP mRNA. STC-1 cells, a cell line derived from a mouse neuroendocrine tumor, were found to express high levels of GIP mRNA. GIP-specific transcripts were not detected in other cell lines tested, which included cells of intestinal, salivary, and endocrine origin. Analysis of GIP-luciferase fusions identified two promoters, a distal and a proximal promoter, upstream of the translation initiation codon for GIP. The distal promoter, located upstream of position +1, corresponds to the principal promoter of the GIP gene and can promote cell specific transcription. Sequential deletion and site-directed mutational analysis of the distal promoter demonstrated that the sequence between -193 and -182 determines cell-specific expression of GIP. Contained in this region is a consensus GATA motif, suggesting that a member of the GATA family of DNA-binding proteins is involved in the cell-specific regulation of the GIP gene. PMID- 9211886 TI - The role of platelet-activating factor-dependent transacetylase in the biosynthesis of 1-acyl-2-acetyl-sn-glycero-3-phosphocholine by stimulated endothelial cells. AB - Acyl analogs of platelet-activating factor (PAF) (1-acyl-2-acetyl-sn-glycero-3 phosphocholine, acylacetyl -GPC) are the predominant products synthesized during thrombin or ionophore A23187-mediated activation of endothelial cells. However, the biosynthetic pathway responsible for the production of acylacetyl-GPC is not well understood. In the present investigation, we have demonstrated that the acyl analogs of PAF are also the major products from calf pulmonary artery endothelial cells in response to a time-dependent stimulation of ATP (10(-3) M), bradykinin (10(-8) M), or ionophore A23187 (2 microM). In addition, we have found that the CoA-independent PAF:acyllyso-GPC transacetylase recently identified by us is concurrently and transiently induced with maximal 4-fold enhancement at 5 min and returned to near basal level by 10 min treatment of endothelial cells with ATP. Acid phosphatase reduces the increased PAF:acyllyso-GPC transacetylase activity from the homogenates of ATP-activated endothelial cells. Reduced PAF:acyllyso-GPC transacetylase activity can be restored by incubating the acid phosphatase treated homogenates with ATP (5 mM) and Mg2+ (10 mM). Furthermore, okadaic acid, a protein phosphatase 1 and 2A inhibitor, incubated with endothelial cells in a dose-dependent manner (1-100 nM) for 10-min potentiates and sustained the stimulation of PAF:acyllyso-GPC transacetylase activity by ATP. On the other hand, genistein, tyrphostin-25 (inhibitors of tyrosine-specific protein kinase), and calphostin C (an inhibitor of protein kinase C) block the activation of PAF:acyllyso-GPC transacetylase by ATP. These results are consistent with the notion that ATP regulates the transacetylase activity by reversible activation and inactivation via the phosphorylation and dephosphorylation cycle. ATP also augments the activities of alkyllyso-GPC/acyllyso-GPC:acetyl-CoA acetyltransferase. However, the activation of the acetyltransferases precedes that of the transacetylase with peak activation occurring at 1-2 min of the ATP treatment. In addition, sodium vanadate, also an inhibitor of protein phosphatase, stimulates the increase in the incorporation of [3H]acetate into acyl[3H]acetyl-GPC of the ATP-treated endothelial cells. Collectively, our data show that both acetyltransferases and transacetylase participate in and contribute to the biosynthesis of acyl analogs of PAF in a coordinate fashion in endothelial cells. PMID- 9211888 TI - Regulation of transducin GTPase activity by human retinal RGS. AB - The intrinsic GTPase activity of transducin controls inactivation of the effector enzyme, cGMP phosphodiesterase (PDE), during turnoff of the visual signal. The inhibitory gamma-subunit of PDE (Pgamma), an unidentified membrane factor and a retinal specific member of the RGS family of proteins have been shown to accelerate GTP hydrolysis by transducin. We have expressed a human homologue of murine retinal specific RGS (hRGSr) in Escherichia coli and investigated its role in the regulation of transducin GTPase activity. As other RGS proteins, hRGSr interacted preferentially with a transitional conformation of the transducin alpha-subunit, GtalphaGDPAlF4-, while its binding to GtalphaGTPgammaS or GtalphaGDP was weak. hRGSr and Pgamma did not compete for the interaction with GtalphaGDPAlF4-. Affinity of the Pgamma-GtalphaGDPAlF4- interaction was modestly enhanced by addition of hRGSr, as measured by a fluorescence assay of GtalphaGDPAlF4- binding to Pgamma labeled with 3-(bromoacetyl)-7 diethylaminocoumarin (PgammaBC). Binding of hRGSr to GtalphaGDPAlF4- complexed with PgammaBC resulted in a maximal approximately 40% reduction of BC fluorescence allowing estimation of the hRGSr affinity for GtalphaGDPAlF4- (Kd 35 nM). In a single turnover assay, hRGSr accelerated GTPase activity of transducin reconstituted with the urea-stripped rod outer segment (ROS) membranes by more than 10-fold to a rate of 0.23 s-1. Addition of Pgamma to the reconstituted system reduced the GTPase level accelerated by hRGSr (kcat 0.085 s-1). The GTPase activity of transducin and the PDE inactivation rates in native ROS membranes in the presence of hRGSr were elevated 3-fold or more regardless of the membrane concentrations. In ROS suspensions containing 30 microM rhodopsin these rates exceeded 0.7 s-1. Our data suggest that effects of hRGSr on transducin's GTPase activity are attenuated by Pgamma but independent of a putative membrane GTPase activating protein factor. The rate of transducin GTPase activity in the presence of hRGSr is sufficient to correlate it with in vivo turnoff kinetics of the visual cascade. PMID- 9211890 TI - Loss of folic acid exporter function with markedly augmented folate accumulation in lipophilic antifolate-resistant mammalian cells. AB - We previously described a 1,000-fold pyrimethamine-resistant Chinese hamster ovary cell line (PyrR100) which retains parental dihydrofolate reductase activity and methotrexate (MTX) sensitivity. This study characterizes the basis for the 14 fold decrease in folic acid and leucovorin concentrations required for clonogenic growth of PyrR100 cells relative to parental AA8 cells. Under conditions in which folic acid reduction was blocked by trimetrexate, PyrR100 cells displayed relative to parental AA8 cells a: 1) 17- and 5-fold increase in the net transport of folic acid and MTX, respectively; 2) 23- and 5-fold decrease in the efflux rate constant for folic acid and MTX, respectively; and 3) 2-fold increase in folic acid influx with no significant change in MTX influx. The markedly increased net folic acid transport in PyrR100 cells could not be explained by cellular folic acid binding, mitochondrial sequestration, polyglutamylation, nor by a decreased membrane potential. The effect of energy deprivation on folic acid and MTX transport in both cell lines was quite different. Glucose and pyruvate deprivation nearly abolished the increase in net folic acid transport in PyrR100 cells. In contrast, energy deprivation increased net MTX transport in AA8 cells, whereas no change was seen with PyrR100 cells. Furthermore, while folic acid influx in PyrR100 and AA8 cells was markedly reduced with energy deprivation, MTX influx was not affected. Provision of glucose and pyruvate to energy-deprived cells resulted in a rapid onset of MTX efflux from parental AA8 cells but not from PyrR100 cells. Taken together these results indicate that the markedly enhanced net transport of folic acid and MTX in PyrR100 cells is largely due to the complete loss of exit pump activity. Furthermore, the energy source that sustains the augmented levels of folic acid appears linked to the influx process and is distinct from the energy source that sustains MTX gradients under these conditions. We conclude that the loss of folic acid efflux is an efficient means of augmenting cellular uptake of folate cofactors and subsequent survival on picomolar folate concentrations. This constitutes the first demonstration of the loss of folic acid exporter activity in mammalian cells as a response to lipophilic antifolate selective pressure. PMID- 9211889 TI - The cytoplasmic domain of granulocyte-macrophage colony-stimulating factor (GM CSF) receptor alpha subunit is essential for both GM-CSF-mediated growth and differentiation. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) regulates differentiation, survival, and proliferation of colony-forming unit-granulocyte macrophage progenitor cells. The biologic actions of GM-CSF are mediated by binding to a specific receptor consisting of two chains designated as alpha and beta subunits. We have demonstrated that the murine FDC-P1-derived cell line WT 19 transfected with the human GM-CSF receptor alpha and beta subunits (GM CSFRalpha and beta) can be induced to differentiate by the addition of human GM CSF (hGM-CSF). By expressing a series of GM-CSFRalpha mutants in WT19 cells, we have determined the amino acid domains of the GM-CSFRalpha cytoplasmic domain that regulate cell differentiation, proliferation, and survival. We found that the membrane proximal proline-rich domain and adjacent 16 residues are essential for both hGM-CSF-dependent cell proliferation and differentiation. In contrast, the C-terminal region of the GM-CSFRalpha cytoplasmic domain was not necessary for cell differentiation mediated by hGM-CSF, but the removal of this region severely impaired the ability of hGM-CSF to support cell survival. While the activation of JAK2, Shc, Erk, and STAT5 proteins correlated with hGM-CSF-mediated cell growth, cellular differentiation occurred in the absence of activation of these signal transduction pathways. PMID- 9211891 TI - Ca2+ binding to the first epidermal growth factor module of coagulation factor VIIa is important for cofactor interaction and proteolytic function. AB - Epidermal growth factor-like (EGF) domain Ca2+ binding sites in the homologous coagulation factors VII, IX, and X stabilize the structural orientation of the gamma-carboxyglutamic acid-rich (Gla) domain relative to EGF-1. Site-directed mutagenesis was employed here to analyze the functional importance of Ca2+ binding to EGF-1 in factor VIIa (VIIa), which initiates coagulation in complex with its cofactor, tissue factor (TF). Ala replacements for Asp63 or Gln49 resulted in reduced TF affinity concordant with the number of eliminated Ca2+ coordinating oxygen atoms in the respective side chains. Ca2+ binding to EGF-1 had no major direct effect on contacts with TF residue Gln110 or on interactions of VIIa residues Arg79 and Phe40, suggesting that the stabilized Gla-EGF-1 orientation affects overall docking. Gly, Ala, and Glu replacements at Asp46, which is a Ca2+-coordinating residue at the Gla aromatic stack carboxyl terminus, are consistent with the notion that an increased flexibility of the Gla domain relative to EGF-1 contributes significantly to loss of function. Certain mutants in the EGF-1 Ca2+ site had reduced proteolytic function, suggesting the importance of the high affinity Ca2+ binding site for macromolecular substrate interaction. PMID- 9211892 TI - Carboxymethylethanolamine, a biomarker of phospholipid modification during the maillard reaction in vivo. AB - Nepsilon-(Carboxymethyl)lysine (CML) is a stable chemical modification of proteins formed from both carbohydrates and lipids during autoxidation reactions. We hypothesized that carboxymethyl lipids such as (carboxymethyl)phosphatidylethanolamine (carboxymethyl-PE) would also be formed in these reactions, and we therefore developed a gas chromatography-mass spectrometry assay for quantification of carboxymethylethanolamine (CME) following hydrolysis of phospholipids. In vitro, CME was formed during glycation of dioleoyl-PE under air and from linoleoylpalmitoyl-PE, but not from dioleoyl PE, in the absence of glucose. In vivo, CME was detected in lipid extracts of red blood cell membranes, approximately 0.14 mmol of CME/mol of ethanolamine, from control and diabetic subjects, (n = 22, p >> 0.5). Levels of CML in erythrocyte membrane proteins were approximately 0.2 mmol/mol of lysine for both control and diabetic subjects (p >> 0.5). For this group of diabetic subjects there was no indication of increased oxidative modification of either lipid or protein components of red cell membranes. CME was also detected in fasting urine at 2-3 nmol/mg of creatinine in control and diabetic subjects (p = 0.085). CME inhibited detection of advanced glycation end product (AGE)-modified protein in a competitive enzyme-linked immunosorbent assay using an anti-AGE antibody previously shown to recognize CML, suggesting that carboxymethyl-PE may be a component of AGE lipids detected in AGE low density lipoprotein. Measurement of levels of CME in blood, tissues, and urine should be useful for assessing oxidative damage to membrane lipids during aging and in disease. PMID- 9211893 TI - The Drosophila ribosomal protein S3 contains a DNA deoxyribophosphodiesterase (dRpase) activity. AB - The Drosophila ribosomal protein S3 has been previously demonstrated to cleave DNA containing 8-oxoguanine residues and has also been found to contain an associated apurinic/apyrimidinic (AP) lyase activity that cleaves phosphodiester bonds via a beta, delta-elimination reaction. The activity of this protein on DNA substrates containing incised AP sites was examined. A glutathione S-transferase fusion protein of S3 was found to efficiently remove sugar-phosphate residues from DNA substrates containing 5'-incised AP sites as well as from DNA substrates containing 3'-incised sites. Removal of 2-deoxyribose-5-phosphate as 4-hydroxy-2 pentenal-5-phosphate from a substrate containing 5'-incised AP sites occurred via a beta-elimination reaction, as indicated by reaction of the released sugar phosphate products with sodium thioglycolate. The reaction for the removal of 4 hydroxy-2-pentenal-5-phosphate from the substrate containing 3'-incised AP sites was dependent on the presence of the Mg2+ cation. These findings suggest that the S3 ribosomal protein may function in several steps of the DNA base excision repair pathway in eukaryotes and may represent an important DNA repair function for the repair of oxidative and ionizing radiation-induced DNA damage. PMID- 9211894 TI - Androgenic induction of prostate-specific antigen gene is repressed by protein protein interaction between the androgen receptor and AP-1/c-Jun in the human prostate cancer cell line LNCaP. AB - In exploring the possible mechanisms of androgen independence of prostate specific antigen (PSA) gene expression, we investigated the effect of elevating AP-1 by both 12-O-tetradecanoylphorbol 13-acetate (TPA) treatment and transfection of the c-Jun expression vector in LNCaP cells. Transcription of PSA is initiated when ligand-activated androgen receptor (AR) binds to a region in the PSA promoter that contains an androgen-responsive element (ARE). It was found that TPA inhibited androgen-induced PSA gene expression by a mechanism that did not alter nuclear levels of AR protein. Overexpression of AP-1 (jun and fos proteins) also inhibited androgen-induced PSA promoter activity. These observations were apparently related to the disruption of AR.ARE complexes as demonstrated by the results of electrophoretic mobility shift assays. Specifically, c-Jun inhibited the formation of AR.ARE complexes and conversely that AR-glutathione S-transferase proteins inhibited the formation of c-Jun.TPA responsive element (TRE) complexes. Consistent with the inhibitory effect of both proteins, anti-c-Jun antibody blocked the inhibition of AR.ARE complex formation by c-Jun. A similar, but less marked, effect was obtained when anti-AR antibody was used to prevent AR inhibition of c-Jun.TRE complex formation. These findings together with results obtained from co-immunoprecipitation experiments strongly suggest that mutual repression of DNA binding activity is due to direct interaction between the two proteins and that the degree of repression may be determined by the ratio of AR to c-Jun. The mechanism of repression studied in mutant analysis experiments yielded evidence of an interaction between the DNA- and ligand-binding domains of AR and the leucine zipper region of c-Jun. Thus, the AR is similar to other nuclear receptors in its ability to interact with AP 1. This association provides a link between AP-1 and AR signal transduction pathways and may play a role in the regulation of the androgen-responsive PSA gene. PMID- 9211895 TI - The N terminus of the Qcr7 protein of the cytochrome bc1 complex is not essential for import into mitochondria in Saccharomyces cerevisiae but is essential for assembly of the complex. AB - Subunit 7 of the yeast cytochrome bc1 complex is encoded by the nuclear QCR7 gene and is essential for respiration. This protein does not contain a cleavable N terminal mitochondrial targeting sequence, and it is not understood how the Qcr7 protein is imported into mitochondria and assembled into the complex. To test the role of the N terminus of the Qcr7 protein in mitochondrial import, assembly of the complex, and proton translocation, we inactivated the endogenous QCR7 gene and expressed mutated qcr7 genes capable of synthesizing proteins truncated by 7, 10, 14, and 20 residues (Qcr7p-delta7, Qcr7p-delta10, Qcr7p-delta14, and Qcr7p delta20, respectively) from the N terminus. In addition, we studied two mutants containing Qcr7 proteins with point mutations in addition to a delta7 truncation, Qcr7p-delta7(D13V) and Qcr7p-delta7(R10K). All the mutant proteins with the exception of Qcr7p-delta10 were present in the mitochondria at 30 degrees C, although most at lower steady-state levels than the Qcr7p from the strain overexpressing wild type QCR7. The absence of the Qcr7p-delta10 may be the result of an unstable protein or a decrease in the efficiency of mitochondrial import due to its compromised amphipathic alpha-helix and the presence of a negative charge exposed at the N terminus. Cytochrome c reductase activities and the amounts of ATP synthesized were comparable with the wild type in the strain expressing Qcr7p-delta7. The strain expressing Qcr7p-delta7(R10K) had an identical phenotype to the one containing the Qcr7p-delta7, whereas strains expressing the Qcr7p-delta10, Qcr7p-delta14, Qcr7p-delta20, and Qcr7p delta7(D13V) were all respiration-deficient. Examination of the steady-state levels of complex III subunits showed that core protein 2, cytochrome c1, the iron-sulfur protein, and the 11-kDa subunit are reduced in respiration-deficient mutant strains. Results from deletion analyses indicate that the N-terminal 20 residues (after Met-1) of the Qcr7 protein are not essential for import into mitochondria and that the N-terminal seven residues (after Met-1) are not involved in proton translocation. The results of this work show, however, that the N terminus of the Qcr7 protein is essential for the biosynthesis of ubiquinol cytochrome c reductase. PMID- 9211896 TI - The RNA molecule CsrB binds to the global regulatory protein CsrA and antagonizes its activity in Escherichia coli. AB - The RNA-binding protein CsrA (carbon storage regulator) is a new kind of global regulator, which facilitates specific mRNA decay. A recombinant CsrA protein containing a metal-binding affinity tag (CsrA-H6) was purified to homogeneity and authenticated by N-terminal sequencing, matrix-assisted laser desorption/ionization time of flight mass spectrometry, and other studies. This protein was entirely contained within a globular complex of approximately 18 CsrA H6 subunits and a single approximately 350-nucleotide RNA, CsrB. cDNA cloning and nucleotide sequencing revealed that the csrB gene is located downstream from syd in the 64-min region of the Escherichia coli K-12 genome and contains no open reading frames. The purified CsrA-CsrB ribonucleoprotein complex was active in regulating glg (glycogen biosynthesis) gene expression in vitro, as was the RNA free form of the CsrA protein. Overexpression of csrB enhanced glycogen accumulation in E. coli, a stationary phase process that is repressed by CsrA. Thus, CsrB RNA is a second component of the Csr system, which binds to CsrA and antagonizes its effects on gene expression. A model for regulatory interactions in Csr is presented, which also explains previous observations on the homologous system in Erwinia carotovora. A highly repeated nucleotide sequence located within predicted stem-loops and other single-stranded regions of CsrB, CAGGA(U/A/C)G, is a plausible CsrA-binding element. PMID- 9211897 TI - The carboxyl-terminal hydrophobic residues of apolipoprotein A-I affect its rate of phospholipid binding and its association with high density lipoprotein. AB - We performed a series of mutations in the human apolipoprotein A-I (apoA-I) gene designed to alter specific amino acid residues and domains implicated in lecithin:cholesterol acyltransferase (LCAT) activation or lipid binding. We used the mutant apoA-I forms to establish nine stable cell lines, and developed strategies for the large scale production and purification of the mutated apoA-I proteins from conditioned media. HDL and dimyristoyl phosphatidylcholine binding assays using the variant apoA-I forms have shown that replacement of specific carboxyl-terminal hydrophobic residues Leu222, Phe225, and Phe229 with lysines, as well as replacement of Leu211, Leu214, Leu218, and Leu219 with valines, diminished the ability of apoA-I to bind to HDL and to lyse dimyristoyl phosphatidylcholine liposomes. The findings indicate that Leu222, and Phe225, Phe229 located in the putative random coil region, and Leu211, Leu214, Leu218, and Leu219 located in the putative helix 8, are important for lipid binding. In contrast, substitutions of alanines for specific charged residues in putative helices 7, 8, or 9 as well as various point mutations in other regions of apoA-I, did not affect the ability of the variant apoA-I forms to bind to HDL or to lyse dimyristoyl phosphatidylcholine liposomes. Cross-linking experiments confirmed that the carboxyl-terminal domain of apoA-I participates in the self-association of the protein, as demonstrated by the inability of the carboxyl-terminal deletion mutants delta185-243 and delta209-243 to form higher order aggregates in solution. Lecithin:cholesterol acyltransferase analysis, using reconstituted HDL particles prepared by the sodium cholate dialysis method, has shown that mutants (Pro165-->Ala,Gln172-->Glu) (Leu211-->Val,Leu214-->Val, Leu218-->Val,Leu219- >Val), Leu222-->Lys,Phe225-->Lys, Phe229-->Lys) and delta209-243 reduced LCAT activation (38-68%). Mutant (Glu191-->Ala,His193-->Ala,Lys195-->Ala) enhanced LCAT activation (131%), and mutant (Ala152-->Leu, Leu159-->Trp) exhibited normal LCAT activation as compared with the wild type proapoA-I and plasma apoA-I forms [corrected]. The apparent catalytic efficiency (Vmax(app)/Km(app)) of the apoA-I mutants ranged from 17.8 to 107.2% of the control and was the result of variations in both the Km and the Vmax in the different mutants. These findings indicate that putative helices 6 and 7, and the carboxyl-terminal helices 8 and 9 contribute to the optimum activation of lecithin:cholesterol acyltransferase. In addition to their use in the present study, the variant apoA-I forms generated will serve as valuable reagents for the identification of the domains and residues of apoA-I involved in binding the scavenger receptor BI, and facilitating cholesterol efflux from cells as well as aid in the structural analysis of apoA-I. PMID- 9211898 TI - Calcium protects a mesophilic xylanase from proteinase inactivation and thermal unfolding. AB - Crystal structure analysis of Pseudomonas fluorescens subsp. cellulosa xylanase A (XYLA) indicated that the enzyme contained a single calcium binding site that did not exhibit structural features typical of the EF-hand motif. Isothermal titration calorimetry revealed that XYLA binds calcium with a Ka of 4.9 x 10(4) M 1 and a stoichiometry consistent with one calcium binding site per molecule of enzyme. Occupancy of the calcium binding domain with its ligand protected XYLA from proteinase and thermal inactivation and increased the melting temperature of the enzyme from 60.8 to 66.5 degrees C. However, the addition of calcium or EDTA did not influence the catalytic activity of the xylanase. Replacement of the calcium binding domain, which is located within loop 7 of XYLA, with the corresponding short loop from Cex (a Cellulomonas fimi xylanase/exoglucanase), did not significantly alter the biochemical properties of the enzyme. These data suggest that the primary function of the calcium binding domain is to increase the stability of the enzyme against thermal unfolding and proteolytic attack. To understand further the nature of the calcium binding domain of XYLA, four variants of the xylanase, D256A, N261A, D262A, and XYLA"', in which Asp-256, Asn 261, and Asp-262 had all been changed to alanine, were constructed. These mutated enzymes did not show any significant binding to Ca2+, indicating that Asp-256, Asn-261, and Asp-262 play a pivotal role in the affinity of XYLA for the divalent cation. In the presence or absence of calcium, XYLA"' exhibited thermal stability similar to that of the native enzyme bound to Ca2+ ions, although the variant was sensitive to proteinase inactivation. The role of the calcium binding domain in vivo and the possible mechanism by which the domain evolved are discussed. PMID- 9211899 TI - Twist-mediated activation of the NK-4 homeobox gene in the visceral mesoderm of Drosophila requires two distinct clusters of E-box regulatory elements. AB - NK-4, also called msh2 and tinman, encodes a homeodomain transcription factor that is required for the development of the dorsal mesoderm and its derivatives in the Drosophila embryo. Genetic analyses indicate that NK-4 resides downstream of the mesodermal determinant twist, which encodes a basic helix-loop-helix-type transcription factor. However, the regulation of NK-4 by twist remains poorly understood. Using expression assays in cultured cells and transgenic flies, we show that two distinct clusters of E-box regulatory sequences, present upstream of the NK-4 gene, mediate NK-4 expression in the visceral mesoderm. These elements are conserved between the Drosophila melanogaster and Drosophila virilis NK-4 genes and serve as binding sites for Twist (E1 cluster) and NK-4 (E2 cluster) proteins. In cultured cells, Twist and NK-4 binding results in activation of NK-4 gene expression. In transgenic animals, the E1 and E2 clusters are functionally connected, and both elements are required for NK-4 activation in cells of the visceral mesoderm and also for NK-4 repression in cells of the somatic musculature. These results demonstrate that NK-4 is a direct transcriptional target for Twist and its own gene product in visceral mesodermal cells, supporting the idea that twist and NK-4 function in the subdivision of the mesoderm during Drosophila embryogenesis. PMID- 9211900 TI - ArgBP2, a multiple Src homology 3 domain-containing, Arg/Abl-interacting protein, is phosphorylated in v-Abl-transformed cells and localized in stress fibers and cardiocyte Z-disks. AB - Arg and c-Abl represent the mammalian members of the Abelson family of protein tyrosine kinases. A novel Arg/Abl-binding protein, ArgBP2, was isolated using a segment of the Arg COOH-terminal domain as bait in the yeast two-hybrid system. ArgBP2 contains three COOH-terminal Src homology 3 domains, a serine/threonine rich domain, and several potential Abl phosphorylation sites. ArgBP2 associates with and is a substrate of Arg and v-Abl, and is phosphorylated on tyrosine in v Abl-transformed cells. ArgBP2 is widely expressed in human tissues and extremely abundant in heart. In epithelial cells ArgBP2 is located in stress fibers and the nucleus, similar to the reported localization of c-Abl. In cardiac muscle cells ArgBP2 is located in the Z-disks of sarcomeres. These observations suggest that ArgBP2 functions as an adapter protein to assemble signaling complexes in stress fibers, and that ArgBP2 is a potential link between Abl family kinases and the actin cytoskeleton. In addition, the localization of ArgBP2 to Z-disks suggests that ArgBP2 may influence the contractile or elastic properties of cardiac sarcomeres and that the Z-disk is a target of signal transduction cascades. PMID- 9211901 TI - Characterization of CLA-1, a human homologue of rodent scavenger receptor BI, as a receptor for high density lipoprotein and apoptotic thymocytes. AB - Recently, a murine scavenger receptor type B class I (SR-BI) was identified that binds high density lipoprotein (HDL) and mediates the selective uptake of cholesterol esters. The human CD36 and LIMPII analogous-1 (CLA-1) receptor shows high sequence homology with SR-BI, but their functional relationship has not been determined. Transfected cells expressing CLA-1 bound HDL with a Kd of about 35 microg/ml, similar to the Kd for HDL binding to rodent SR-BI. This binding resulted in an intracellular accumulation of HDL-derived [3H]cholesterol esters without internalization or degradation of 125I-apolipoprotein. CLA-1 was strongly expressed in the adrenal gland and was also abundant in liver and testis, suggesting that CLA-1, like SR-BI, could play a role in the metabolism of HDL. However, CLA-1 was also expressed in monocytes and, like SR-BI, recognized modified forms of low density lipoproteins as well as native LDL and anionic phospholipids. These findings suggest that CLA-1 might have additional physiological functions. We found that CLA-1 recognizes apoptotic thymocytes. Our results demonstrate that CLA-1, a close structural homologue of SR-BI, is also functionally related to SR-BI and may play an important role as a "docking receptor" for HDL in connection with selective uptake of cholesterol esters. An additional role in recognition of damaged cells is suggested by these studies. PMID- 9211902 TI - Identification and reconstruction of the binding site within alphaMbeta2 for a specific and high affinity ligand, NIF. AB - Engagement of the alphaMbeta2 (CD11b/CD18, Mac-1) integrin on neutrophils supports adhesion and induces various cellular responses. These responses can be blocked by a specific ligand of alphaMbeta2, neutrophil inhibitory factor (NIF). The molecular basis of alphaMbeta2-NIF interactions was studied. The single chain alphaM subunit, expressed on the surface of human 293 cells, bound NIF with an affinity equivalent to that of alphaMbeta2 heterodimer. This observation, coupled with previous data showing that the alphaMI domain alone supported high affinity NIF binding, indicated that the binding site for NIF is restricted to the I domain. Guided by the crystal structure of the alphaMI domain, 16 segments corresponding to the entire outer hydrated surface of alphaMI domain were switched to their counterparts sequences in alphaL, which does not bind NIF. Surface expression and heterodimer formation were achieved for all mutants, and correct folding was confirmed. Of the 16 switches, only 5 affected NIF binding substantially, reducing affinity by 8-300-fold. These data confined the NIF binding site to a narrow region composed of Pro147-Arg152, Pro201-Lys217, and Asp248-Arg261 of alphaM. Verifying this localization, when these segments were introduced into the alphaXI-domain, the resulting chimeric receptor was converted into a high affinity NIF-binding protein. PMID- 9211903 TI - Stress, apoptosis, and mitosis induce phosphorylation of human keratin 8 at Ser 73 in tissues and cultured cells. AB - Simple epithelia express keratins 8 (K8) and 18 (K18) as their major intermediate filament proteins. We previously showed that several types of cell stress such as heat and virus infection result in a distinct hyperphosphorylated form of K8 (termed HK8). To better characterize K8/18 phosphorylation, we generated monoclonal antibodies by immunizing mice with hyperphosphorylated keratins that were purified from colonic cultured human HT29 cells pretreated with okadaic acid. One antibody specifically recognized HK8, and the epitope was identified as 71LLpSPL which corresponds to K8 phosphorylation at Ser-73. Generation of HK8 occurs in mitotic HT29 cells, basal crypt mitotic cells in normal mouse intestine, and in regenerating mouse hepatocytes after partial hepatectomy. Prominent levels of HK8 were also generated in HT29 cells that were induced to undergo apoptosis using anisomycin or etoposide. In addition, mouse hepatotoxicity that is induced by chronic feeding with griseofulvin resulted in HK8 formation in the liver. Our results demonstrate that a "reverse immunological" approach, coupled with enhancing in vivo phosphorylation using phosphatase inhibitors, can result in the identification of physiologic phosphorylation states. As such, K8 Ser-73 phosphorylation generates a distinct HK8 species under a variety of in vivo conditions including mitosis, apoptosis, and cell stress. The low steady state levels of HK8 during mitosis, in contrast to stress and apoptosis, suggest that accumulation of HK8 may represent a physiologic stress marker for simple epithelia. PMID- 9211904 TI - Crystal structures of substrate binding site mutants of manganese peroxidase. AB - Manganese peroxidase (MnP), an extracellular heme enzyme from the lignin degrading basidiomycetous fungus, Phanerochaete chrysosporium, catalyzes the oxidation of MnII to MnIII. The latter, acting as a diffusible redox mediator, is capable of oxidizing a variety of lignin model compounds. The proposed MnII binding site of MnP consists of a heme propionate, three acidic ligands (Glu-35, Glu-39, and Asp-179), and two water molecules. Using crystallographic methods, this binding site was probed by altering the amount of MnII bound to the protein. Crystals grown in the absence of MnII, or in the presence of EDTA, exhibited diminished electron density at this site. Crystals grown in excess MnII exhibited increased electron density at the proposed binding site but nowhere else in the protein. This suggests that there is only one major MnII binding site in MnP. Crystal structures of a single mutant (D179N) and a double mutant (E35Q,D179N) at this site were determined. The mutant structures lack a cation at the MnII binding site. The structure of the MnII binding site is altered significantly in both mutants, resulting in increased access to the solvent and substrate. PMID- 9211905 TI - Kinetic analysis of four HIV-1 reverse transcriptase enzymes mutated in the primer grip region of p66. Implications for DNA synthesis and dimerization. AB - The highly conserved primer grip region in the p66 subunit of HIV-1 reverse transcriptase (RT) is formed by the beta12-beta13 hairpin (residues 227-235). It has been proposed to play a role in aligning the 3'-OH end of the primer in a position for nucleophilic attack on an incoming dNTP. To analyze the importance of the primer grip for RT function, mutant RTs were used that contain single alanine substitutions of residues Trp229, Met230, Gly231, and Tyr232 in the p66 subunit of the heterodimeric p66/51 enzyme. Steady-state and pre-steady-state kinetic analyses of the enzymes were performed. All mutant enzymes revealed reduced polymerase activity. Mutation of Y232A showed the smallest effect on polymerase function. Equilibrium fluorescence titrations demonstrated that the affinity of the mutants for tRNA was only slightly affected. However, the affinity for primer-template DNA was reduced 27-fold for mutant p66(W229A)/51 and 23-fold for mutant p66(G231A)/51, and the maximal pre-steady-state rate of nucleotide incorporation, kpol, was reduced 27-fold for p66(W229A)/51 and 70-fold for p66(G231A)/51, respectively. Mutant p66(M230A)/51 revealed no reduced affinity for primer-template but showed a 71-fold reduced affinity for dTTP. Additionally, the mutations Trp229 and Gly231 affected the stability of the RT heterodimer. PMID- 9211907 TI - Oxidation process of bovine heart ubiquinol-cytochrome c reductase as studied by stopped-flow rapid-scan spectrophotometry and simulations based on the mechanistic Q cycle model. AB - Stopped-flow rapid-scan spectrophotometry was employed to study complicated oxidation processes of ubiquinol-cytochrome c reductase (QCR) that was purified from bovine heart mitochondria and maximally contained 0.36 mol of ubiquinone 10/mol of heme c1. When fully reduced QCR was allowed to react with dioxygen in the presence of cytochrome c plus cytochrome c oxidase, the oxidation of b-type hemes accompanied an initial lag, apparently low potential heme bL was oxidized first, followed by high potential heme bH. Antimycin A inhibited the oxidation of both b-type hemes. The oxidation of heme c1 was triphasic and became biphasic in the presence of antimycin A. On the other hand, starting from partially reduced QCR that was poised at a higher redox potential with succinate and succinate cytochrome c reductase, the b-type hemes were oxidized immediately without a lag. When the ubiquinone content in QCR was as low as 0.1 mol/mol heme c1 the oxidation of the b-type hemes was almost suppressed. As the Q-deficient QCR was supplemented with ubiquinol-2, the rapid oxidation of b-type hemes was restored to some extent. These results indicate that a limited amount of ubiquinone-10 found in purified preparations of QCR is obligatory for electron transfer from the b-type hemes to iron-sulfur protein (ISP) and heme c1. The characteristic oxidation profiles of heme bL, heme bH, and heme c1 were simulated successfully based on a mechanistic Q cycle model. According to the simulations the two electron oxidation of ubiquinol-10 via the ISP and heme c1 pathway, which is more favorable thermodynamically than the bifurcation of electron flow into both ISP and heme bL, does really occur as long as heme bL is in the reduced state and provides ubiquinone-10 at center i. Mechanistically this process takes time, thus explaining the initial lag in the oxidation of the b-type hemes. With the partially reduced QCR, inherent ubisemiquinone at center i immediately oxidizes reduced heme bH thus eliminating the lag. The mechanistic Q cycle model consists of 56 reaction species, which are interconnected by the reaction paths specified with microscopic rate constants. The simulations further indicate that the rate constants for electron transfer between the redox centers can be from 10(5) to 10(3) s-1 and are rarely rate-limiting. On the other hand, a shuttle of ubiquinone or ubiquinol between center o and center i and the oxidation of heme c1 can be rate-limiting. The interplay of the microscopic rate constants determines the actual reaction pathway that is shown schematically by the "reaction map." Most significantly, the simulations support the consecutive oxidation of ubiquinol in center o as long as both heme bL and heme bH are in the reduced state. Only when heme bL is oxidized and ISP is reduced can SQo donate an electron to heme bL. Thus, we propose that a kinetic control mechanism, or "a kinetic switch," is significant for the bifurcation of electron flow. PMID- 9211906 TI - Amino acid limitation induces expression of CHOP, a CCAAT/enhancer binding protein-related gene, at both transcriptional and post-transcriptional levels. AB - In mammals, plasma concentrations of amino acids are affected by nutritional or pathological conditions. Here we examined the role of amino acid limitation in regulating the expression of CHOP, a CCAAT/enhancer binding protein (C/EBP) related gene. CHOP protein is capable of interacting with other C/EBPs to modify their DNA binding activities and may function as a negative regulator of these transcription factors. Our data show that leucine limitation in human cell lines leads to induction of CHOP mRNA and protein in a dose-dependent manner. CHOP mRNA induction is rapidly reversed by leucine replenishment. Elevated mRNA levels result from both an increase in the rate of CHOP transcription and an increase in the CHOP mRNA stability. Using a transient expression assay, we show that a promoter fragment, when linked to a reporter gene, is sufficient to mediate the regulation of CHOP expression by leucine starvation in HeLa cells. In addition, we found that decreasing amino acid concentration by itself can induce CHOP expression independently of a cellular stress due to protein synthesis inhibition. Moreover, CHOP expression is induced at leucine concentrations in the range of those observed in blood of protein-restricted animals suggesting that amino acids can participate, in concert with hormones, in the regulation of gene expression. PMID- 9211908 TI - Two modes of ligand binding in maltose-binding protein of Escherichia coli. Correlation with the structure of ligands and the structure of binding protein. AB - Ligands that are transported by the maltose transport system of Escherichia coli must first bind to the periplasmic maltose-binding protein (MBP). However, binding of a ligand does not always lead to its transport. As reported earlier, reduced or oxidized maltodextrins bind tightly to MBP but are not transported; some mutant MBPs, such as MalE254, bind maltodextrins tightly but cannot produce their transport. In this study, UV differential spectroscopy and fluorescence emission spectroscopy were used to study the modes by which various ligands bind to MBP. Maltose binding produced a red shift in the fluorescence emission spectrum of wild type MBP and a sharp hypochromatic trend below 265 nm in its UV spectrum (R mode (for red)). On the other hand, binding of reduced, oxidized, or cyclic maltodextrins produced a pronounced blue shift in the fluorescence emission spectrum of wild type MBP and a peak at about 250 nm in its UV difference spectrum (B mode (for blue). Binding of reducing maltodextrins to wild type MBP produced spectral changes that seemed to be a mixture of predominantly R mode binding and some B mode binding, whereas their binding to mutant MBP MalE254 produced changes indicative of pure B mode binding. Thus, the ligands that are bound exclusively via the B mode to either the wild type or MalE254 MBP are not transported. PMID- 9211909 TI - Two modes of ligand binding in maltose-binding protein of Escherichia coli. Electron paramagnetic resonance study of ligand-induced global conformational changes by site-directed spin labeling. AB - Binding of ligands to the maltose-binding protein (MBP) of Escherichia coli often causes a global conformational change involving the closure of its two lobes. We have introduced a cysteine residue onto each of these lobes by site-directed mutagenesis and modified these residues with spin labels. Using EPR spectroscopy, we examined the changes, caused by the ligand binding, in distance between the two spin labels, hence between the two lobes. The binding of both maltose and maltotetraose induced a considerable closure of the N- and C-terminal lobes of MBP. Little closure occurred upon the binding of maltotetraitol or beta cyclodextrin. Previous study by fluorescence and UV differential absorbance spectroscopy (Hall, J. A., Gehring, K., and Nikaido, H. (1997) J. Biol. Chem. 272, 17605-17609) showed that maltose and a large portion of maltotetraose bound to MBP via one mode (R mode or "end-on" mode), which is physiologically active and leads to the subsequent transport of the ligands across the cytoplasmic membrane. In contrast, maltotetraitol and beta-cyclodextrin bound to MBP via a different mode (B mode or "middle" mode), which is physiologically inactive. The present work suggests that the B mode is nonproductive because ligands binding in this manner prevent the closure of the two domains of MBP, and, as a result, the resulting ligand-MBP complex is incapable of interacting properly with the inner membrane-associated transporter complex. PMID- 9211910 TI - Two modes of ligand binding in maltose-binding protein of Escherichia coli. Functional significance in active transport. AB - In the preceding two papers (Hall, J. A., Gehring, K., and Nikaido, H. (1997) J. Biol. Chem. 272, 17605-17609; Hall, J. A., Thorgeirson, T. E., Liu, J., Shin, Y. E., and Nikaido, H. (1997) J. Biol. Chem. 272, 17610-17614), we showed that ligands that bind to the Escherichia coli maltose-binding protein (MBP) without producing the closure of its two lobes are not transported into the cytoplasm. Here, we examine various combinations of ligands, MBPs, and membrane-associated transporters, by utilizing reconstituted proteoliposomes, right side-out membrane vesicles, and intact cells. Closed forms of wild type MBP, complexed with maltose or maltodextrins, interacted with wild type transporter complex to stimulate the hydrolysis of ATP by MalK ATPase located on the other side of the membrane, as shown earlier for the maltose-MBP complex (Davidson, A. L., Shuman, H. A., and Nikaido, H. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 2360-2364). In contrast, open forms of liganded MBPs, such as the complex containing wild type MBP and reduced, oxidized, or cyclic maltodextrins or the complex containing the mutant MBP MalE254 and unmodified maltodextrins, did not stimulate ATP hydrolysis, suggesting that the proper interaction between the ligand-MBP complex and the external surface of the transporter requires the former to be in the closed conformation. However, when a mutant transporter containing MalG511 was used, the already significant basal level of ATP hydrolysis was further stimulated not only by ligand MBPs in the closed form but also by those in the open form (except that containing beta-cyclodextrin), data suggesting that the mutant transporter does not always require the closed MBP complex presumably because of its exceptionally strong affinity to MBP, described earlier (Dean, D. A., Hor, L.-I., Shuman, H. A., and Nikaido, H. (1992) Mol. Microbiol. 6, 2033-2040). Furthermore, this mutant transporter was able to transport reduced maltodextrin, and cells expressing the transporter were able to grow by using reduced maltodextrin, if the periplasmic concentrations of MBP were kept low so as not to inhibit the transport process. PMID- 9211911 TI - Androgen-dependent protein interactions within an intron 1 regulatory region of the 20-kDa protein gene. AB - The 20-kDa protein gene is androgen regulated in rat ventral prostate. Intron 1 contains a 130-base pair complex response element (D2) that binds androgen (AR) and glucocorticoid receptor (GR) but transactivates only with AR in transient cotransfection assays in CV1 cells using the reporter vector D2-tkCAT. To better understand the function of this androgen-responsive unit, nuclear protein interactions with D2 were analyzed by DNase I footprinting in ventral prostate nuclei of intact or castrated rats and in vitro with ventral prostate nuclear protein extracts from intact, castrated, and testosterone-treated castrated rats. Multiple androgen-dependent protected regions and hypersensitive sites were identified in the D2 region with both methods. Mobility shift assays with 32P labeled oligonucleotides spanning D2 revealed specific interactions with ventral prostate nuclear proteins. Four of the D2-protein complexes decreased in intensity within 24 h of castration. UV cross-linking of the androgen-dependent DNA binding proteins identified protein complexes of approximately 140 and 55 kDa. The results demonstrate androgen-dependent nuclear protein-DNA interactions within the complex androgen response element D2. PMID- 9211912 TI - Mmot1, a new helix-loop-helix transcription factor gene displaying a sharp expression boundary in the embryonic mouse brain. AB - Several genetic factors have been proven to contribute to the specification of the metencephalic-mesencephalic territory, a process that sets the developmental foundation for prospective morphogenesis of the cerebellum and mesencephalon. However, evidence stemming from genetic and developmental studies performed in man and various model organisms suggests the contribution of many additional factors in determining the fine subdivision and differentiation of these central nervous system regions. In man, the cerebellar ataxias/aplasias represent a large and heterogeneous family of genetic disorders. Here, we describe the identification by differential screening and the characterization of Mmot1, a new gene encoding a DNA-binding protein strikingly similar to the helix-loop-helix factor Ebf/Olf1. Throughout midgestation embryogenesis, Mmot1 is expressed at high levels in the metencephalon, mesencephalon, and sensory neurons of the nasal cavity. In vitro DNA binding data suggest some functional equivalence of Mmot1 and Ebf/Olf1, possibly accounting for the reported lack of olfactory or neural defects in Ebf-/- knockout mutants. The isolation of Mmot1 and of an additional homolog in the mouse genome defines a novel, phylogenetically conserved mammalian family of transcription factor genes of potential relevance in studies of neural development and its aberrations. PMID- 9211913 TI - A nuclear localization signal of human aryl hydrocarbon receptor nuclear translocator/hypoxia-inducible factor 1beta is a novel bipartite type recognized by the two components of nuclear pore-targeting complex. AB - Aryl hydrocarbon receptor nuclear translocator (ARNT) is a component of the transcription factors, aryl hydrocarbon receptor (AhR) and hypoxia-inducible factor 1, which transactivate their target genes, such as CYP1A1 and erythropoietin, in response to xenobiotic aromatic hydrocarbons and to low O2 concentration, respectively. Since ARNT was isolated as a factor required for the nuclear translocation of AhR from the cytoplasm in response to xenobiotics, the subcellular localization of ARNT has been of great interest. In this investigation, we analyzed the subcellular distribution of ARNT using transient expression of a fusion gene with beta-galactosidase and microinjection of recombinant proteins containing various fragments of ARNT in the linker region of glutathione S-transferase/green fluorescent protein. We found a clear nuclear localization of ARNT in the absence of exogenous ligands to AhR, and identified the nuclear localization signal (NLS) of amino acid residues 39-61. The characterized NLS consists of 23 amino acids, and can be classified as a novel variant of the bipartite type on the basis of having two separate regions responsible for efficient nuclear translocation activity, but considerable deviation of the sequence from the consensus of the classical bipartite type NLSs. Like the well characterized NLS of the SV40 T-antigen, this variant bipartite type of ARNT NLS was also mediated by the two components of nuclear pore targeting complex, PTAC58 and PTAC97, to target to the nuclear rim in an in vitro nuclear transport assay. PMID- 9211914 TI - The mechanism of preferential degradation of polyadenylated RNA in the chloroplast. The exoribonuclease 100RNP/polynucleotide phosphorylase displays high binding affinity for poly(A) sequence. AB - Polyadenylation of mRNA in the chloroplast has recently been shown to target the RNA molecule for rapid exonucleolytic degradation. A model has been suggested in which the degradation of chloroplast mRNA is initiated by endonucleolytic cleavage(s) followed by the addition of poly(A)-rich sequences and rapid exonucleolytic degradation. When in vitro transcribed RNAs were incubated with chloroplast protein extract, competition between polyadenylated and non polyadenylated RNAs for degradation resulted in the rapid degradation of the polyadenylated molecules and stabilization of their non-polyadenylated counterparts. To elucidate the molecular mechanism governing this effect, we determined whether the chloroplast exoribonuclease 100RNP/polynucleotide phosphorylase (PNPase) preferably degrades polyadenylated RNA. When separately incubated with each molecule, isolated 100RNP/PNPase degraded polyadenylated and non-polyadenylated RNAs at the same rate. However, when both molecules were mixed together, the polyadenylated RNA was degraded, whereas the non-polyadenylated RNA was stabilized. In RNA binding experiments, 100RNP/PNPase bound the poly(A) sequence with much higher affinity than other RNA molecules, thereby defining the poly(A)-rich RNA as a preferential substrate for the enzyme. 100RNP/PNPase may therefore be involved in a mechanism in which post-transcriptional addition of poly(A)-rich sequence targets the chloroplast RNA for rapid exonucleolytic degradation. PMID- 9211915 TI - In vitro chromatin assembly of the HIV-1 promoter. ATP-dependent polar repositioning of nucleosomes by Sp1 and NFkappaB. AB - Nuclease hypersensitive sites exist in vivo in the chromatin of the integrated human immunodeficiency virus (HIV)-1 proviral genome, in the 5'-long terminal repeat (LTR) within the promoter/enhancer region near Sp1 and NFkappaB binding sites. Previous studies from the Kadonaga and Jones laboratories have shown that Sp1 and NFkappaB can establish hypersensitive sites in a truncated form of this LTR when added before in vitro chromatin assembly with Drosophila extracts, thus facilitating subsequent transcriptional activation of a linked reporter gene upon the association of additional factors (Pazin, M. J., Sheridan, P. L., Cannon, K., Cao, Z., Keck, J. G., Kadanaga, J. T., and Jones, K. A. (1996) Genes & Dev. 10, 37-49). Here we assess the role of a full-length LTR and 1 kilobase pair of downstream flanking HIV sequences in chromatin remodeling when these transcription factors are added after chromatin assembly. Using Xenopus laevis oocyte extracts to assemble chromatin in vitro, we have confirmed that Sp1 and NFkappaB can indeed induce sites hypersensitive to DNase I, micrococcal nuclease, or restriction enzymes on either side of factor binding sites in chromatin but not naked DNA. We extend these earlier studies by demonstrating that the process is ATP-dependent when the factors are added after chromatin assembly and that histone H1, AP1, TBP, or Tat had no effect on hypersensitive site formation. Furthermore, we have found that nucleosomes upstream of NFkappaB sites are rotationally positioned prior to factor binding and that their translational frame is registered after binding NFkappaB. On the other hand, binding of Sp1 positions adjacent downstream nucleosome(s). We term this polar repositioning because each factor aligns nucleosomes only on one side of its binding sites. Mutational analysis and oligonucleotide competition each demonstrated that this remodeling required Sp1 and NFkappaB binding sites. PMID- 9211916 TI - Purification and characterization of CobT, the nicotinate-mononucleotide:5,6 dimethylbenzimidazole phosphoribosyltransferase enzyme from Salmonella typhimurium LT2. AB - We report the purification and biochemical characterization of the cobalamin biosynthetic enzyme nicotinate-mononucleotide:5, 6-dimethylbenzimidazole phosphoribosyltransferase (CobT) from Salmonella typhimurium. cobT was overexpressed and the protein purified to approximately 97% homogeneity. NH2 terminal sequence analysis confirmed that the protein encoded by cobT was purified. Homogeneous CobT catalyzed the synthesis of N1-(5-phospho-alpha-D ribosyl)-5,6-dimethylbenzimidazole. The identity of high performance liquid chromatography-purified product was confirmed by fast atom bombardment mass spectrometry. CobT activity was optimal at 45 degrees C and pH 10.0. The apparent Km for nicotinate mononucleotide was 680 microM; the apparent Km for 5, 6 dimethylbenzimidazole was less than 10 microM. CobT used nicotinamide mononucleotide as a ribose phosphate donor. CobT phosphoribosylated alternative base substrates including benzimidazole, 4,5-dimethyl-1,2-phenylenediamine, imidazole, histidine, adenine, and guanine in vitro. The resulting ribotides were incorporated into cobamides that were differentially utilized by methionine synthase (EC 2.1.1.13), ethanolamine ammonia-lyase (EC 4.3.1.7), and 1,2 propanediol dehydratase (EC 4.2.1.28) in vivo. The lack of base substrate specificity by CobT may explain the inability to isolate mutants blocked in the synthesis of 5, 6-dimethylbenzimidazole in this bacterium. PMID- 9211917 TI - Mutational analysis of putative SCH 28080 binding sites of the gastric H+,K+ ATPase. AB - A compound, SCH 28080 (2-methyl-8-(phenylmethoxy)imidazo [1,2-a]pyridine-3 acetonitrile), reversibly inhibits gastric and renal ouabain-insensitive H+,K+ ATPase, but not colonic ouabain-sensitive H+,K+-ATPase. By using the functional expression system and site-directed mutagenesis, we analyzed the putative binding sites of SCH 28080 in gastric H+,K+-ATPase alpha-subunit. It was previously reported that the binding site of SCH 28080, which is a K+-site inhibitor specific for gastric H+,K+-ATPase, was in the first extracellular loop between the first and second transmembrane segments of the alpha-subunit; Phe-126 and Asp 138 were putative binding sites. However, we found that all the mutants in the first extracellular loop including Phe-126 and Asp-138 retained H+, K+-ATPase activity and sensitivity to SCH 28080. Therefore, amino acid residues in the first extracellular loop are not directly involved in the SCH 28080 binding nor indispensable for the H+, K+-ATPase activity. Here we propose a candidate residue that is important for the binding with SCH 28080, Glu-822 in the sixth transmembrane segment. Mutations of Glu-822 to Asp and Ala (mutants termed E822D and E822A, respectively) decreased the ATPase activity to about 45% and 35% of the wild-type enzyme, respectively, while the mutations to Gln and Leu abolished the activity. Mutant E822A showed a significantly lower affinity for K+ than the wild-type enzyme, indicating that Glu-822 is involved in determining the affinity for K+. The sensitivity of mutant E822D to SCH 28080 was 8 times lower than that of the wild-type enzyme. The counterpart of Glu-822 in gastric H+,K+-ATPase is Asp in Na+,K+-ATPase and other colonic ouabain-sensitive H+,K+-ATPase, which are insensitive to SCH 28080. These results suggest that Glu-822 is one of important sites that bind with SCH 28080. PMID- 9211918 TI - RecA as a motor protein. Testing models for the role of ATP hydrolysis in DNA strand exchange. AB - ATP hydrolysis (by RecA protein) fundamentally alters the properties of RecA protein-mediated DNA strand exchange reactions. ATP hydrolysis renders DNA strand exchange unidirectional, greatly increases the lengths of hybrid DNA created, permits the bypass of heterologous DNA insertions in one or both DNA substrates, and is absolutely required for exchange reactions involving four DNA strands. There are at least two viable models to explain how ATP hydrolysis is coupled to DNA strand exchange so as to bring about these effects. The first couples ATP hydrolysis to a redistribution of RecA monomers within a RecA filament. The second couples ATP hydrolysis to a facilitated rotation of the DNA substrates. The RecA monomer redistribution model makes the prediction that heterology bypass should not occur if the single-stranded DNA substrate is linear. The facilitated DNA rotation model predicts that RecA protein should promote the separation of paired DNA strands within a RecA filament if one of them is contiguous with a length of DNA being rotated about the filament exterior. Here, a facile bypass of heterologous insertions with linear DNA substrates is demonstrated, providing evidence against a role for RecA monomer redistribution in heterology bypass. In addition, we demonstrate that following a four-strand DNA exchange reaction, a distal segment of DNA hundreds of base pairs in length can be unwound in a nonreciprocal phase of the reaction, consistent with the direct coupling of an ATP hydrolytic motor to the proposed DNA rotation. PMID- 9211919 TI - Beta3-adrenergic receptors on white and brown adipocytes mediate beta3-selective agonist-induced effects on energy expenditure, insulin secretion, and food intake. A study using transgenic and gene knockout mice. AB - beta3-Adrenergic receptors (beta3-ARs) are expressed predominantly on white and brown adipocytes, and acute treatment of mice with CL 316,243, a potent and highly selective beta3-AR agonist, produces a 2-fold increase in energy expenditure, a 50-100-fold increase in insulin levels, and a 40-50% reduction in food intake. Recently, we generated gene knockout mice lacking functional beta3 ARs and demonstrated that each of these responses were mediated exclusively by beta3-ARs. However, the tissue site responsible for producing these actions is unknown. In the present study, genetically engineered mice were created in which beta3-ARs are expressed exclusively in white and brown adipocytes (WAT+BAT-mice), or in brown adipocytes only (BAT-mice). This was accomplished by injecting tissue specific beta3-AR transgenic constructs into mouse zygotes homozygous for the beta3-AR knockout allele. Control, knockout, WAT+BAT, and BAT-mice were then treated acutely with CL, and the effects on various parameters were assessed. As previously observed, all effects of CL were completely absent in gene knockout mice lacking beta3-ARs. The effects on O2 consumption, insulin secretion, and food intake were completely rescued with transgenic re-expression of beta3-ARs in white and brown adipocytes (WAT+BAT-mice), demonstrating that each of these responses is mediated exclusively by beta3-ARs in white and/or brown adipocytes, and that beta3-ARs in other tissue sites were not required. Importantly, transgenic re-expression of beta3-ARs in brown adipocytes only (BAT-mice) failed to rescue, in any way, CL-mediated effects on insulin levels and food intake and only minimally restored effects on oxygen consumption, indicating that any effect on insulin secretion and food intake, and a full stimulation of oxygen consumption required the presence of beta3-ARs in white adipocytes. The mechanisms by which beta3-AR agonist stimulation of white adipocytes produces these responses are unknown but may involve novel mediators not previously known to effect these processes. PMID- 9211920 TI - Interaction of growth hormone-activated STATs with SH2-containing phosphotyrosine phosphatase SHP-1 and nuclear JAK2 tyrosine kinase. AB - Growth hormone (GH) rapidly stimulates tyrosine phosphorylation followed by serine/threonine phosphorylation of multiple cytoplasmic STAT transcription factors, including one, STAT5b, that is uniquely responsive to the temporal pattern of plasma GH stimulation in rat liver and is proposed to play a central role in the activation of male-expressed liver genes by GH pulses in vivo (Waxman, D. J., Ram, P. A., Park, S. H., and Choi, H. K. (1995) J. Biol. Chem. 270, 13262-13270). We now show that JAK2, the GH receptor-associated tyrosine kinase, is present both in the cytosol and in the nucleus in cultured liver cells and in rat liver in vivo and that GH-activated STAT3 but not STAT5b becomes associated with nuclear JAK2. GH is also shown to activate by 3-4-fold SHP-1, a phosphotyrosine phosphatase that contains two src homology 2 (SH2) domains. GH also induces nuclear translocation and binding of SHP-1 to tyrosine phosphorylated STAT5b, suggesting that this GH-activated phosphatase may play a role in dephosphorylation leading to deactivation of nuclear STAT5b following the termination of a plasma GH pulse in male rat liver in vivo. No such association of SHP-1 with GH-activated STAT3 was detected, a finding that could help explain the marked desensitization of STAT3, but not STAT5b, to subsequent GH pulses following an initial GH activation event. PMID- 9211921 TI - Evolution of RNA-binding specificity in T4 DNA polymerase. AB - DNA polymerase of phage T4 (T4 gp43), an essential component of the T4 DNA replicase, is a multifunctional single-chained (898-amino acid) protein that catalyzes the highly accurate synthesis of DNA in phage replication. The enzyme functions both as a DNA-binding replication protein and as a sequence-specific RNA-binding autogenous translational repressor. We have utilized a phylogenetic approach to study the relationships between the two nucleic acid-binding functions of the protein. We found that autogenous translational control of gp43 biosynthesis has been conserved in phage RB69, a distant relative of T4, although we also found that the RB69 system differs from its T4 counterpart in two regards: (a) nucleotide sequence and predicted secondary structure of the RNA target (translational operator), and (b) RNA specificity of the protein. T4 gp43 is specific to the RNA operator sequence of the T4 genome whereas RB69 gp43 can bind and repress operator RNA from both phages equally well. In studies with T4 RB69 gp43 chimeras, we mapped T4 gp43 RNA-binding specificity to a protein segment that also harbors important determinants for DNA binding and the polymerase catalytic function. Our results suggest that RNA functions as a regulator of both the dosage and activity of this DNA replication enzyme. PMID- 9211922 TI - Homeostatic regulation of copper uptake in yeast via direct binding of MAC1 protein to upstream regulatory sequences of FRE1 and CTR1. AB - Copper deprivation of Saccharomyces cerevisiae induces transcription of the FRE1 and CTR1 genes. FRE1 encodes a surface reductase capable of reducing and mobilizing copper chelates outside the cell, and CTR1 encodes a protein mediating copper uptake at the plasma membrane. In this paper, the protein encoded by MAC1 is identified as the factor mediating this homeostatic control. A novel dominant allele of MAC1, MAC1(up2), is mutated in a Cys-rich domain that may function in copper sensing (a G to A change of nucleotide 812 resulting in a Cys-271 to Tyr substitution). This mutant is functionally similar to the MAC1(up1) allele in which His-279 in the same domain has been replaced by Gln. Both mutations confer constitutive copper-independent expression of FRE1 and CTR1. A sequence including the palindrome TTTGCTCA ... TGAGCAAA, appearing within the 5'-flanking region of the CTR1 promoter, is necessary and sufficient for the copper- and MAC1-dependent CTR1 transcriptional regulation. An identical sequence appears as a direct repeat in the FRE1 promoter. The data indicate that the signal resulting from copper deprivation is transduced via the Cys-rich motif of MAC1 encompassing residues 264-279. MAC1 then binds directly and specifically to the CTR1 and FRE1 promoter elements, inducing transcription of those target genes. This model defines the homeostatic mechanism by which yeast regulates the cell acquisition of copper in response to copper scarcity or excess. PMID- 9211923 TI - Amidophosphoribosyltransferase limits the rate of cell growth-linked de novo purine biosynthesis in the presence of constant capacity of salvage purine biosynthesis. AB - Factors controlling relative flux rates of the de novo and salvage pathways of purine nucleotide biosynthesis during animal cell growth are not fully understood. To examine the relative role of each pathway for cell growth, three cell lines including CHO K1 (a wild-type Chinese hamster ovary fibroblast cell line), CHO ade -A (an auxotrophic cell line deficient of amidophosphoribosyltransferase (ATase), a presumed rate-limiting enzyme of the de novo pathway), and CHO ade -A transfected with human ATase cDNA (-A+hATase) resulting in 30-350% of the ATase activity of CHO K1, were cultured in purine rich or purine-free media. Based on the enzyme activities of ATase and hypoxanthine phosphoribosyltransferase, the metabolic rate of the de novo and salvage pathways, the rate of cell growth (growth rate) in three cell lines under various culture conditions, and the effect of hypoxanthine infusion on the metabolic rate of the de novo pathway in rat liver, we concluded the following. 1) In -A+hATase transfectants, ATase activity limits the rate of the de novo pathway, which is closely linked with the growth rate. 2) Purine nucleotides are synthesized preferentially by the salvage pathway as long as hypoxanthine, the most essential source of purine salvage, can be utilized, which was confirmed in rat liver in vivo by hypoxanthine infusion. The preferential usage of the salvage pathway results in sparing the energy expenditure required for de novo synthesis. 3) The regulatory capacity of the de novo pathway (about 200%) was larger than that of the salvage pathway (about 20%) with constant hypoxanthine phosphoribosyltransferase activity. PMID- 9211924 TI - Phosphorylation of Na,K-ATPase by protein kinase C at Ser18 occurs in intact cells but does not result in direct inhibition of ATP hydrolysis. AB - Na,K-ATPase activity has been demonstrated to be regulated by a variety of hormones in different tissues. It is known to be directly phosphorylated on its alpha-subunit, but the functional effects of protein kinases remain controversial. We have developed a sensitive, antibody-based assay for detection of the level of phosphorylation of the alpha1-isoform of rat Na,K-ATPase at the serine residue that is most readily phosphorylated by protein kinase C (PKC) in vitro, Ser18. By stimulation of endogenous PKC and inhibition of phosphatase activity, it was possible to consistently obtain a very high stoichiometry of phosphorylation (close to 0.9) in several types of intact cells. This demonstrates the accessibility and competency of the site for endogenous phosphorylation. The cells used were derived from rat (NRK 52E, C6, L6, and primary cultures of cerebellar granule cells, representing epithelial cells, glia, muscle cells, and neurons). In the presence of the phosphatase inhibitor calyculin A, full phosphorylation was preserved during subsequent assays of enzyme activity in vitro. Assay of the hydrolysis of ATP in NRK and C6 cells, however, indicated that there was no significant effect of phosphorylation on the Vmax of the Na, K-ATPase or on the apparent affinity for Na+. Any regulatory effect of PKC on sodium pump activity thus must be lost upon disruption or permeabilization of the cells and is not a direct consequence of enzyme alteration by covalent phosphorylation of Ser18. PMID- 9211925 TI - Phosphorylation and desensitization of human endothelin A and B receptors. Evidence for G protein-coupled receptor kinase specificity. AB - Although endothelin-1 can elicit prolonged physiologic responses, accumulating evidence suggests that rapid desensitization affects the primary G protein coupled receptors mediating these responses, the endothelin A and B receptors (ETA-R and ETB-R). The mechanisms by which this desensitization proceeds remain obscure, however. Because some intracellular domain sequences of the ETA-R and ETB-R differ substantially, we tested the possibility that these receptor subtypes might be differentially regulated by G protein-coupled receptor kinases (GRKs). Homologous, or receptor-specific, desensitization occurred within 4 min both in the ETA-R-expressing A10 cells and in 293 cells transfected with either the human ETA-R or ETB-R. In 293 cells, this desensitization corresponded temporally with agonist-induced phosphorylation of each receptor, assessed by receptor immunoprecipitation from 32Pi-labeled cells. Agonist-induced receptor phosphorylation was not substantially affected by PKC inhibition but was reduced 40% (p << 0.03) by GRK inhibition, effected by a dominant negative GRK2 mutant. Inhibition of agonist-induced phosphorylation abrogated agonist-induced ETA-R desensitization. Overexpression of GRK2, -5, or -6 in 293 cells augmented agonist induced ET-R phosphorylation approximately 2-fold (p << 0.02), but each kinase reduced receptor-promoted phosphoinositide hydrolysis differently. While GRK5 inhibited ET-R signaling by only approximately 25%, GRK2 inhibited ET-R signaling by 80% (p << 0.01). Congruent with its superior efficacy in suppressing ET-R signaling, GRK2, but not GRK5, co-immunoprecipitated with the ET-Rs in an agonist dependent manner. We conclude that both the ETA-R and ETB-R can be regulated indistinguishably by GRK-initiated desensitization. We propose that because of its affinity for ET-Rs demonstrated by co-immunoprecipitation, GRK2 is the most likely of the GRKs to initiate ET-R desensitization. PMID- 9211926 TI - Dynamic targeting of the agonist-stimulated m2 muscarinic acetylcholine receptor to caveolae in cardiac myocytes. AB - In cardiac myocytes, as well as specialized conduction and pacemaker cells, agonist binding to muscarinic acetylcholine receptors (mAchRs) results in the activation of several signal transduction cascades including the endothelial isoform of nitric-oxide synthase (eNOS) expressed in these cells. Recent evidence indicates that, as in endothelial cells, eNOS in cardiac myocytes is localized to plasmalemma caveolae, specialized lipid microdomains that contain caveolin-3, a muscle-specific isoform of the scaffolding protein caveolin. In this report, using a detergent-free method for isolation of sarcolemmal caveolae from primary cultures of adult rat ventricular myocytes, we demonstrated that the muscarinic cholinergic agonist carbachol promotes the translocation of mAchR into low density gradient fractions containing most myocyte caveolin-3 and eNOS. Following isopycnic centrifugation, the different gradient fractions were exposed to the muscarinic radioligand [3H]quinuclidinyl benzilate (QNB), and binding was determined after membrane filtration or immunoprecipitation. In a direct radioligand binding assay, we found that [3H]QNB binding can be detected in caveolin-enriched fractions only when cardiac myocytes have been previously exposed to carbachol. Furthermore, most of this [3H]QNB binding can be specifically immunoprecipitated by an antibody to the m2 mAchR, indicating that the translocation of this receptor subtype is responsible for the [3H]QNB binding detected in the low density fractions. Moreover, the [3H]QNB binding could be quantitatively immunoprecipitated from the light membrane fractions with a caveolin-3 antibody (but not a control IgG1 antibody), confirming that the m2 mAchR is targeted to caveolae after carbachol treatment. Importantly, atropine, a muscarinic cholinergic antagonist, did not induce translocation of m2 mAchR to caveolae and prevented receptor translocation in response to the agonist carbachol. Thus, dynamic targeting of sarcolemmal m2 mAchR to caveolae following agonist binding may be essential to initiate specific downstream signaling cascades in these cells. PMID- 9211927 TI - Two protein-tyrosine phosphatases inactivate the osmotic stress response pathway in yeast by targeting the mitogen-activated protein kinase, Hog1. AB - Protein phosphatases inactivate mitogen-activated protein kinase (MAPK) signaling pathways by dephosphorylating components of the MAPK cascade. Two genes encoding protein-tyrosine phosphatases, PTP2, and a new phosphatase, PTP3, have been isolated in a genetic selection for negative regulators of an osmotic stress response pathway called HOG, for high osmolarity glycerol, in budding yeast. PTP2 and PTP3 were isolated as multicopy suppressors of a severe growth defect due to hyperactivation of the HOG pathway. Phosphatase activity is required for suppression since mutation of the catalytic Cys residue in Ptp2 and Ptp3, destroys suppressor function and biochemical activity. The substrate of these phosphatases is likely to be the MAPK, Hog1. Catalytically inactive Ptp2 and Ptp3 coprecipitate with Hog1 from yeast extracts. In addition, strains lacking PTP2 and PTP3 do not dephosphorylate Hog1-phosphotyrosine as well as wild type. The latter suggests that PTP2 and PTP3 play a role in adaptation. Consistent with this role, osmotic stress induces expression of PTP2 and PTP3 transcripts in a Hog1-dependent manner. Thus Ptp2 and Ptp3 likely act in a negative feedback loop to inactivate Hog1. PMID- 9211928 TI - Phosphatidylinositol-4-phosphate 5-kinase isozymes catalyze the synthesis of 3 phosphate-containing phosphatidylinositol signaling molecules. AB - Phosphatidylinositol-4-phosphate 5-kinases (PIP5Ks) utilize phosphatidylinositols containing D-3-position phosphates as substrates to form phosphatidylinositol 3,4 bisphosphate. In addition, type I PIP5Ks phosphorylate phosphatidylinositol 3, 4 bisphosphate to phosphatidylinositol 3,4,5-trisphosphate, while type II kinases have less activity toward this substrate. Remarkably, these kinases can convert phosphatidylinositol 3-phosphate to phosphatidylinositol 3,4,5-trisphosphate in a concerted reaction. Kinase activities toward the 3-position phosphoinositides are comparable with those seen with phosphatidylinositol 4-phosphate as the substrate. Therefore, the PIP5Ks can synthesize phosphatidylinositol 4,5 bisphosphate and two 3-phosphate-containing polyphosphoinositides. These unexpected activities position the PIP5Ks as potential participants in the generation of all polyphosphoinositide signaling molecules. PMID- 9211929 TI - Architecture of the yeast cell wall. Beta(1-->6)-glucan interconnects mannoprotein, beta(1-->)3-glucan, and chitin. AB - In a previous study (Kollar, R., Petrakova, E., Ashwell, G., Robbins, P. W., and Cabib, E. (1995) J. Biol. Chem. 270, 1170-1178), the linkage region between chitin and beta(1-->3)-glucan was solubilized and isolated in the form of oligosaccharides, after digestion of yeast cell walls with beta(1-->3)-glucanase, reduction with borotritide, and subsequent incubation with chitinase. In addition to the oligosaccharides, the solubilized fraction contained tritium-labeled high molecular weight material. We have now investigated the nature of this material and found that it represents areas in which all four structural components of the cell wall, beta(1-->3)-glucan, beta(1-->6)-glucan, chitin, and mannoprotein are linked together. Mannoprotein, with a protein moiety about 100 kDa in apparent size, is attached to beta(1-->6)-glucan through a remnant of a glycosylphosphatidylinositol anchor containing five alpha-linked mannosyl residues. The beta(1-->6)-glucan has some beta(1-->3)-linked branches, and it is to these branches that the reducing terminus of chitin chains appears to be attached in a beta(1-->4) or beta(1-->2) linkage. Finally, the reducing end of beta(1-->6)-glucan is connected to the nonreducing terminal glucose of beta(1- >3)-glucan through a linkage that remains to be established. A fraction of the isolated material has three of the main components but lacks mannoprotein. From these results and previous findings on the linkage between mannoproteins and beta(1-->6)-glucan, it is concluded that the latter polysaccharide has a central role in the organization of the yeast cell wall. The possible mechanism of synthesis and physiological significance of the cross-links is discussed. PMID- 9211930 TI - Ykt6p, a prenylated SNARE essential for endoplasmic reticulum-Golgi transport. AB - Vesicular transport between secretory compartments requires specific recognition molecules called SNAREs. Here we report the identification of three putative SNAREs, p14 (Sft1p), p28 (Gos1p), and a detailed characterization of p26 (Ykt6p). All three were originally isolated as interacting partners of the cis Golgi target membrane-associated SNARE Sed5p, when Sec18p (yeast NSF) was inactivated. YKT6 is an essential gene that codes for a novel vesicle-associated SNARE functioning at the endoplasmic reticulum-Golgi transport step in the yeast secretory pathway. Depletion of Ykt6p results in the accumulation of the p1 precursor (endoplasmic reticulum form) of the vacuolar enzyme carboxypeptidase Y and morphological abnormalities consistent with a defect in secretion. Membrane localization of Ykt6p is essential for protein function and is normally mediated by isoprenylation. However, replacement of the isoprenylation motif with a bona fide transmembrane anchor results in a functional protein confirming that membrane localization, but not isoprenylation per se, is required for function. Ykt6p and its homologues are highly conserved from yeast to human as demonstrated by the functional complementation of the loss of Ykt6p by its human counterpart. This is the first example of a human SNARE protein functionally replacing a yeast SNARE. This observation implies that the specific details of the vesicle targeting code, like the genetic code, are conserved in evolution. PMID- 9211932 TI - Functional equivalence of creatine kinase isoforms in mouse skeletal muscle. AB - Creatine kinase (CK) is a highly conserved enzyme abundant in skeletal muscle that has a key role in high energy phosphate metabolism. The localization of the muscle isoenzyme of CK (MM-CK) to the M line and the sarcoplasmic reticulum of myofibrils has been suggested to be important for proper force development in skeletal muscle. The importance of this subcellular compartmentation has not been directly tested in vivo. To test the role of myofibrilar localization of CK, the consequences of a complete CK isoform switch from MM-CK to the brain (BB-CK) isoform, which does not localize to the M line, was studied in transgenic mouse skeletal muscle. In MM-CK knockout mice there are large contractile defects. When MM-CK was replaced by BB-CK, the aberrant contractile phenotypes seen in MM-CK knockout mice were returned to normal despite the lack of myofibrillar localization. These results indicate that CK compartmentation to the myofibril of skeletal muscle is not essential for contractile function and that there is functional equivalence of creatine kinase isoforms in supporting cellular energy metabolism. PMID- 9211931 TI - Interactions of the human mitochondrial protein import receptor, hTom20, with precursor proteins in vitro reveal pleiotropic specificities and different receptor domain requirements. AB - Tom20 is part of a multiple component, dynamic complex that functions to import specific cytosolic proteins into or through the outer membrane of the mitochondrion. To analyze the contribution of Tom20 to precursor protein recognition, the cytosolic domain of the human mitochondrial import receptor, hTom20, has been expressed as a fusion protein with glutathione S-transferase and conditions established to measure specific interactions of the receptor component with precursor proteins in vitro. Reconstitution of receptor binding from purified components revealed that a prototypic matrix-destined precursor protein, pODHFR, interacts with Tom20 by a mechanism that is dependent on an active matrix targeting signal but does not require cytosolic components or ATP. Binding was influenced by both salt concentration and detergent. The effect of salt or detergent, however, varied for different precursor proteins. In particular, detergent selectively enhanced binding of pODHFR to receptor, possibly because of induced changes in the structure of the signal sequence. Finally, mutations were introduced into hTom20 which had a dramatic effect on binding of some precursor proteins but not on others. Taken together, the results suggest that hTom20 recognizes and physically interacts with precursor proteins bearing a diverse array of topogenic sequences and that such pleiotropic specificity for these precursor proteins may involve different domains within the receptor molecule. PMID- 9211933 TI - Lipopolysaccharide induction of the tumor necrosis factor-alpha promoter in human monocytic cells. Regulation by Egr-1, c-Jun, and NF-kappaB transcription factors. AB - Biosynthesis of tumor necrosis factor-alpha (TNF-alpha) is predominantly by cells of the monocytic lineage. This study examined the role of various cis-acting regulatory elements in the lipopolysaccharide (LPS) induction of the human TNF alpha promoter in cells of monocytic lineage. Functional analysis of monocytic THP-1 cells transfected with plasmids containing various lengths of TNF-alpha promoter localized enhancer elements in a region (-182 to -37 base pairs (bp)) that were required for optimal transcription of the TNF-alpha gene in response to LPS. Two regions were identified: region I (-182 to -162 bp) contained an overlapping Sp1/Egr-1 site, and region II (-119 to -88) contained CRE and NF kappaB (designated kappaB3) sites. In unstimulated THP-1, CRE-binding protein and, to a lesser extent, c-Jun complexes were found to bind to the CRE site. LPS stimulation increased the binding of c-Jun-containing complexes. In addition, LPS stimulation induced the binding of cognate nuclear factors to the Egr-1 and kappaB3 sites, which were identified as Egr-1 and p50/p65, respectively. The CRE and kappaB3 sites in region II together conferred strong LPS responsiveness to a heterologous promoter, whereas individually they failed to provide transcriptional activation. Furthermore, increasing the spacing between the CRE and the kappaB3 sites completely abolished LPS induction, suggesting a cooperative interaction between c-Jun complexes and p50/p65. These studies indicate that maximal LPS induction of the TNF-alpha promoter is mediated by concerted participation of at least two separate cis-acting regulatory elements. PMID- 9211934 TI - PU.1 is essential for p47(phox) promoter activity in myeloid cells. AB - Expression of the phagocyte cytosolic protein p47(phox), a component of NADPH oxidase, is restricted mainly to myeloid cells. To study the cis-elements and trans-acting factors responsible for its gene expression, we have cloned and characterized the p47(phox) promoter. A predominant transcriptional start site was identified 21 nucleotides upstream of the translation initiation codon. To identify the gene promoter sequences, transient transfections of HL-60 human myeloid cells were performed with a series of 5'-deletion p47(phox)-luciferase reporter constructs that extended as far upstream as -3050 bp relative to the transcriptional start site. The -224 and -86 constructs had the strongest p47(phox) promoter activity, whereas the -46 construct showed a major reduction in activity and the -36 construct a complete loss of activity. DNase I footprint analysis identified a protected region from -37 to -53. This region containing a consensus PU.1 site bound specifically both PU.1 present in nuclear extracts from myeloid cells and PU.1 synthesized in vitro. Mutations of this site eliminated PU.1 binding and abolished the ability of the p47(phox) promoter to direct expression of the reporter gene. The p47(phox) promoter was active in all myeloid cell lines tested (HL-60, THP-1, U937, PLB-985), but not in non-myeloid cells (HeLa, HEK293). Finally, PU.1 trans-activated the p47(phox)-luciferase constructs in HeLa cells. We conclude that, similar to certain other myeloid-specific genes, p47(phox) promoter activity in myeloid cells requires PU.1. PMID- 9211935 TI - Activation of the receptor for advanced glycation end products triggers a p21(ras)-dependent mitogen-activated protein kinase pathway regulated by oxidant stress. AB - Advanced glycation end products (AGEs) exert their cellular effects on cells by interacting with specific cellular receptors, the best characterized of which is the receptor for AGE (RAGE). The transductional processes by which RAGE ligation transmits signals to the nuclei of cells is unknown and was investigated. AGE albumin, a prototypic ligand, activated p21(ras) in rat pulmonary artery smooth muscle cells that express RAGE, whereas nonglycated albumin was without effect. MAP kinase activity was enhanced at concentrations of AGE-albumin, which activated p21(ras) and NF-kappaB. Depletion of intracellular glutathione rendered cells more sensitive to AGE-mediated activation of this signaling pathway. In contrast, signaling was blocked by preventing p21(ras) from associating with the plasma membrane or mutating Cys118 on p21(ras) to Ser. Signaling was receptor dependent, because it was prevented by blocking access to RAGE with either anti RAGE IgG or by excess soluble RAGE. These data suggest that RAGE-mediated induction of cellular oxidant stress triggers a cascade of intracellular signals involving p21(ras) and MAP kinase, culminating in transcription factor activation. The molecular mechanism that triggers this pathway likely involves oxidant modification and activation of p21(ras). PMID- 9211936 TI - Complex formation of the elongation factor Tu from Pseudomonas aeruginosa with nucleoside diphosphate kinase modulates ribosomal GTP synthesis and peptide chain elongation. AB - The elongation factor Tu (EF-Tu) from Pseudomonas aeruginosa was purified as a 45 kDa polypeptide that forms a complex with both the 12- and 16-kDa forms of nucleoside-diphosphate kinase (Ndk) and predominantly synthesizes GTP. 70 S ribosomes of P. aeruginosa predominantly synthesize GTP, which is inhibited in presence of anti-Ndk antibodies. Anti-EF-Tu antibodies change the specificity of ribosomal GTP synthesis to all nucleoside triphosphate synthesis. Ndk has been shown to be a part of 30 S ribosomes, whereas EF-Tu is found to be associated with the 50 S ribosomal subunit. These data indicate that GTP synthesis in the ribosome is modulated both by Ndk and by EF-Tu. Peptide chain elongation as measured by polymerization of Phe-tRNA on a poly(U) template in presence of GDP can be inhibited by anti-Ndk antibodies and restored by the addition of GTP. Anti EF-Tu antibodies similarly inhibit peptide chain elongation by P. aeruginosa ribosomes in the in vitro translation assay; however, this inhibition cannot be overcome by adding back GTP. Because the purified EF-Tu.16-kDa Ndk complex predominantly synthesizes GTP, it seems likely that this complex is a significant source of GTP for translational elongation in protein biosynthesis. PMID- 9211937 TI - Analysis of the psbU gene encoding the 12-kDa extrinsic protein of photosystem II and studies on its role by deletion mutagenesis in Synechocystis sp. PCC 6803. AB - The gene encoding the 12-kDa extrinsic protein of photosystem II from Synechocystis sp. PCC 6803 was cloned based on N-terminal sequence of the mature protein. This gene, named psbU, encodes a polypeptide of 131 residues, the first 36 residues of which were absent in the mature protein and thus served as a transit peptide required for its transport into the thylakoid lumen. A psbU gene deletion mutant grew photoautotrophically in normal BG11 medium at almost the same rate as that of the wild type strain. This mutant, however, grew apparently slower than the wild type did upon depletion of Ca2+ or Cl- from the growth medium. Photosystem II oxygen evolution decreased to 81% in the mutant as compared with that in the wild type, and the thermoluminescence B- and Q-bands shifted to higher temperatures accompanied by an increase in the Q-band intensity. These results indicate that the 12-kDa protein is not essential for oxygen evolution but may play a role in optimizing the ion (Ca2+ and Cl-) environment and maintaining a functional structure of the cyanobacterial oxygen evolving complex. In addition, a double deletion mutant lacking cytochrome c-550 and the 12-kDa protein grew photoautotrophically with a phenotype identical to that of the single deletion mutant of cytochrome c-550. This supports our previous biochemical results that the 12-kDa protein cannot bind to photosystem II in the absence of cytochrome c-550 (Shen, J.-R., and Inoue, Y. (1993) Biochemistry 32, 1825-1832). PMID- 9211938 TI - Interleukin-1 reduces the glycolytic utilization of glucose by pancreatic islets and reduces glucokinase mRNA content and protein synthesis by a nitric oxide dependent mechanism. AB - Culture of rat pancreatic islets with interleukin-1 (IL-1) results in up regulation of the inducible isoform of nitric oxide synthase and overproduction of nitric oxide (NO). This is associated with reversible inhibition of both glucose-induced insulin secretion and islet glucose oxidation, and these effects are prevented by the inducible nitric oxide synthase inhibitor NG monomethylarginine. IL-1 also induces accumulation of nonesterified arachidonic acid in islets by an NO-dependent mechanism, and one potential explanation for that effect would involve an IL-1-induced enhancement of islet glycolytic flux. We have therefore examined effects of IL-1 on islet glycolytic utilization of glucose and find that culture of islets with IL-1 in medium containing 5.5 mM glucose results in suppression of islet glucose utilization subsequently measured at glucose concentrations between 6 and 18 mM. The IL-1-induced suppression of islet glucose utilization is associated with a decline in islet glucokinase mRNA content, as determined by competitive reverse transcriptase-polymerase chain reaction, and in glucokinase protein synthesis, as determined by immuoprecipitation experiments, and all of these effects are prevented by NG monomethylarginine. These findings suggest that IL-1 can down-regulate islet glucokinase, which is the primary component of the islet glucose-sensor apparatus, by an NO-dependent mechanism. Because reductions in islet glucokinase levels are known to cause a form of type II diabetes mellitus, these observations raise the possibility that factors which increase islet NO levels might contribute to development of glucose intolerance. PMID- 9211939 TI - Interaction of arrestins with intracellular domains of muscarinic and alpha2 adrenergic receptors. AB - The intracellular domains of G-protein-coupled receptors provide sites for interaction with key proteins involved in signal initiation and termination. As an initial approach to identify proteins interacting with these receptors and the receptor motifs required for such interactions, we used intracellular subdomains of G-protein-coupled receptors as probes to screen brain cytosol proteins. Peptides from the third intracellular loop (i3) of the M2-muscarinic receptor (MR) (His208-Arg387), M3-MR (Gly308-Leu497), or alpha2A/D-adrenergic receptor (AR) (Lys224-Phe374) were generated in bacteria as glutathione S-transferase (GST) fusion proteins, bound to glutathione-Sepharose and used as affinity matrices to detect interacting proteins in fractionated bovine brain cytosol. Bound proteins were identified by immunoblotting following SDS-polyacrylamide gel electrophoresis. Brain arrestins bound to the GST-M3 fusion protein, but not to the control GST peptide or i3 peptides derived from the alpha2A/D-AR and M2-MR. However, each of the receptor subdomains bound purified beta-arrestin and arrestin-3. The interaction of the M3-MR and M2-MR i3 peptides with arrestins was further investigated. The M3-MR i3 peptide bound in vitro translated [3H]beta arrestin and [3H]arrestin-3, but did not interact with in vitro translated or purified visual arrestin. The properties and specificity of the interaction of in vitro translated [3H]beta-arrestin, [3H]visual arrestin, and [3H]beta arrestin/visual arrestin chimeras with the M2-MR i3 peptide were similar to those observed with the intact purified M2-MR that was phosphorylated and/or activated by agonist. Subsequent binding site localization studies indicated that the interaction of beta-arrestin with the M3-MR peptide required both the amino (Gly308-Leu368) and carboxyl portions (Lys425-Leu497) of the receptor subdomain. In contrast, the carboxyl region of the M3-MR i3 peptide was sufficient for its interaction with arrestin-3. PMID- 9211940 TI - Regulation of expression of the human MTH1 gene encoding 8-oxo-dGTPase. Alternative splicing of transcription products. AB - The enzyme 8-oxo-7,8-dihydrodeoxyguanosine triphosphatase (8-oxo-dGTPase) hydrolyzes 8-oxo-dGTP to 8-oxo-dGMP, thereby preventing misincorporation of 8-oxo dGTP into DNA. We investigated expression of MTH1 gene encoding 8-oxo-dGTPase. Large amounts of MTH1 mRNA were present in thymus and testis, embryonic tissues, and certain cell lines. In peripheral blood lymphocytes, the level of MTH1 mRNA was significantly increased after concomitant treatment with phytohemagglutinin and interleukin-2. Analyses of the 5' regions of the MTH1 transcripts revealed that 7 types of MTH1 mRNAs, which may be produced by transcription initiation at different sites and/or alternative splicing. The MTH1 gene consists of 5 major exons, some of which are composed of differentially processed segments. All types of MTH1 mRNAs carry the entire coding region, and may be functional. Three ATG initiation codons in-frame were found in the 5' regions of some of the MTH1 mRNAs. There is a polymorphic alteration at the 5' splicing site (GT to GC) located in exon 2, an event which affects splicing patterns of the MTH1 transcript. Allele frequency of this polymorphism is about 20% among healthy volunteers. PMID- 9211941 TI - Peptide mapping of the murine DNA methyltransferase reveals a major phosphorylation site and the start of translation. AB - The murine DNA methyltransferase catalyzes the transfer of methyl groups from S adenosylmethionine to cytosines within d(CpG) dinucleotides. The enzyme is necessary for normal embryonic development and is implicated in a number of important processes, including the control of gene expression and cancer. Metabolic labeling and high pressure liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) were performed on DNA methyltransferase purified from murine erythroleukemia cells. Serine 514 was identified as a major phosphorylation site that lies in a domain required for targeting of the enzyme to the replication foci. These results present a potential mechanism for the regulation of DNA methylation. HPLC-ESI-MS peptide mapping data demonstrated that the purified murine DNA methyltransferase protein contains the N-terminal regions predicted by the recently revised 5' gene sequences (Yoder, J. A., Yen, R.-W. C., Vertino, P. M., Bestor, T. H. , and Baylin, S. B. (1996) J. Biol. Chem. 271, 31092-31097). The evidence suggests a start of translation at the first predicted methionine, with no alternate translational start sites. Our peptide mapping results provide a more detailed structural characterization of the DNA methyltransferase that will facilitate future structure/function studies. PMID- 9211942 TI - Bradykinin sequesters B2 bradykinin receptors and the receptor-coupled Galpha subunits Galphaq and Galphai in caveolae in DDT1 MF-2 smooth muscle cells. AB - In this report, we show that the vasoactive peptide agonist bradykinin (BK) when bound to B2 BK receptors on DDT1 MF-2 smooth muscle cells promotes the recruitment and sequestration of the occupied receptors and the receptor-coupled G-protein alpha subunits Galphaq and Galphai in caveolae. Association of ligand receptor complexes and Galpha subunits with caveolae was indicated by their co enrichment on density gradients with caveolin, a marker protein for caveolae. Caveolin and Galpha subunits were monitored by immunoblotting, whereas receptors were monitored as ligand receptor complexes formed by labeling receptors with the agonist BK or the antagonist NPC17731 prior to cell disruption and caveolae enrichment. These complexes were detected with radioligand and by immunoblotting with BK antibodies. A direct interaction of Galpha subunits with caveolin was also indicated by their co-immunoprecipitation. Immunoelectron microscopy revealed that the enriched caveolin, Galpha subunits, and BK receptor complexes were present in structures of 0.1-0.2 microm. At 4 degrees C, BK and NPC17731 receptor complexes were detected in caveolae, and both complexes were sensitive to acid washing prior to cell disruption and caveolae enrichment. Elevation of the temperature to 37 degrees C increased the amount of BK receptor complexes in caveolae with a maximal response at 10 min (continuous labeling) or 20 min (single-round labeling), and the complexes became acid-resistant. These conditions also increased the amount of Galphaq and Galphai in caveolae with a maximal response at 5-10 min. In contrast, the NPC17731 receptor complexes remained acid-sensitive and dissociated at this temperature, and antagonists did not increase the amount of Galpha subunits in caveolae. These results show that some agonists that act through G-protein-coupled receptors promote the association of their receptors and receptor-coupled Galpha subunits with caveolae. PMID- 9211943 TI - The smallest membrane anchoring subunit (QPs3) of bovine heart mitochondrial succinate-ubiquinone reductase. Cloning, sequencing, topology, and Q-binding domain. AB - The cDNA encoding the smallest membrane-anchoring subunit (QPs3) of bovine heart mitochondrial succinate-ubiquinone reductase was cloned and sequenced. This cDNA is 1330 base pairs long with an open reading frame of 474 base pairs that encodes the 103 amino acid residues of mature QPs3 and a 55-amino acid residue presequence. The cDNA insert has an 820-base pair long 3'-untranslated region, including a poly(A) tail. The molecular mass of QPs3, deduced from the nucleotide sequence, is 10,989 Da. QPs3 is a very hydrophobic protein; the hydropathy plot of the amino acid sequence reveals three transmembrane helices. Previous photoaffinity labeling studies of succinate-ubiquinone reductase, using 3-azido-2 methyl-5-methoxy[3H]-6-decyl-1,4-benzoquinone ([3H]azido-Q), identified QPs3 as one of the putative Q-binding proteins in this reductase. An azido-Q-linked peptide with a retention time of 66 min is obtained by high performance liquid chromatography of the chymotrypsin digest of carboxymethylated and succinylated [3H]azido-Q-labeled QPs3 purified from labeled succinate-ubiquinone reductase by a procedure involving phenyl-Sepharose 4B column chromatography, preparative SDS polyacrylamide gel electrophoresis, and acetone precipitation. The amino acid sequence of this peptide is NH2-L-N-P-C-S-A-M-D-Y-COOH, corresponding to residues 29-37. The structure of QPs3 in the inner mitochondrial membrane is proposed based on the hydropathy profile of the amino acid sequence, on the predicted tendencies to form alpha-helices and beta-sheets, and on immunobinding of Fab' fragmenthorseradish peroxidase conjugates prepared from antibodies against two synthetic peptides, corresponding to the NH2 terminus region and the loop connecting helices 2 and 3 of QPs3, in mitoplasts and submitochondrial particles. The ubiquinone-binding domain in the proposed model of QPs3 is probably located at the end of transmembrane helix 1 toward the C-side of the mitochondrial inner membrane. PMID- 9211944 TI - Protein phosphatase 2C acts independently of stress-activated kinase cascade to regulate the stress response in fission yeast. AB - Stress-activated signal transduction pathways, which are largely conserved among a broad spectrum of eukaryotic species, have a crucial role in the survival of many forms of stress. It is therefore important to discover how these pathways are both positively and negatively regulated. Recent genetic studies have implicated protein phosphatase 2C (PP2C) as a novel negative regulator of stress response pathways in both budding and fission yeasts. Moreover, it was hypothesized that PP2C dephosphorylates one or more components of protein kinase cascades that are at the core of stress-activated signal transduction pathways. Herein we present genetic and biochemical studies of the fission yeast Schizosaccharomyces pombe that disprove this hypothesis and indicate that PP2C instead negatively regulates a downstream element of the pathway. First, high expression of PP2C produces phenotypes that are inconsistent with negative regulation of the Wik1-Wis1-Spc1 stress-activated kinase cascade. Second, high expression of PP2C leads to sustained activating tyrosine phosphorylation of Spc1. Third, Spc1-dependent phosphorylation of Atf1, a transcription factor substrate of Spc1, is unaffected by high expression of PP2C. Fourth, high expression of PP2C suppresses Atf1-dependent transcription of a stress-response gene. These studies strongly suggest that PP2C acts downstream of Spc1 kinase in the stress-activated signal transduction pathway. PMID- 9211945 TI - Diversity of the Escherichia coli type 1 fimbrial lectin. Differential binding to mannosides and uroepithelial cells. AB - Type 1 fimbriae are the most common adhesive organelles of Escherichia coli. Because of their virtual ubiquity, previous epidemiological studies have not found a correlation between the presence of type 1 fimbriae and urinary tract infections (UTIs). Recently it has become clear that type 1 fimbriae exhibit several different phenotypes, due to allelic variation of the gene for the lectin subunit, FimH, and that these phenotypes are differentially distributed among fecal and UTI isolates. In this study, we have analyzed in more detail the ability of isogenic, recombinant strains of E. coli expressing fimH genes of the predominant fecal and UTI phenotypes to adhere to glycoproteins and to uroepithelial cells. Evidence was obtained to indicate that type 1 fimbriae differ in their ability to recognize various mannosides, utilizing at least two different mechanisms. All FimH subunits studied to date are capable of mediating adhesion via trimannosyl residues, but only certain variants are capable of mediating high levels of adhesion via monomannosyl residues. The ability of the FimH lectins to interact with monomannosyl residues strongly correlates with their ability to mediate E. coli adhesion to uroepithelial cells. In this way, it would be possible for certain phenotypic variants of type 1 fimbriae to contribute more than others to virulence of E. coli in the urinary tract. PMID- 9211946 TI - Involvement of stress-activated protein kinase and p38/RK mitogen-activated protein kinase signaling pathways in the enhanced phosphorylation of initiation factor 4E in NIH 3T3 cells. AB - The initiation factor (eIF) 4E is regulated by modulating both the phosphorylation and the availability of the protein to participate in the initiation process. Here we show that either serum treatment or activation of the stress-activated protein kinase (JNK/SAPK) led to enhanced phosphorylation of eIF4E in quiescent NIH 3T3 cells. Although the immunosuppressant, rapamycin, was found to stabilize the association of eIF4E with its negative regulator, 4E-BP1, this drug did not prevent the early effects of serum stimulation on the overall rate of translation, polysome formation, the phosphorylation status of eIF4E, or the recruitment of eIF4E into the eIF4F complex. However, the rapid enhancement of eIF4E phosphorylation in response to serum was largely prevented by the inhibitor of mitogen-activated protein (MAP) kinase activation, PD98059. Activation of the JNK/SAPK signaling pathway with anisomycin resulted in enhanced phosphorylation of eIF4E, which was prevented by either rapamycin or the highly specific p38 MAP kinase inhibitor, SB203580. These data illustrate that multiple signaling pathways, including those of distinct members of the MAP kinase family, mediate the phosphorylation of eIF4E and that the association of eIF4E with 4E BP1 does not necessarily prevent phosphorylation of eIF4E in vivo. PMID- 9211947 TI - Pediatric musculoskeletal computed radiography. AB - BACKGROUND: In conventional radiography, a film-screen system serves as the X-ray detector and the film also functions as an archival and display medium. Unlike film-screen radiography, these functions are uncoupled in computed radiography (CR). CR uses conventional radiographic equipment to expose an image on a storage phosphor plate instead of a film-screen combination. OBJECTIVE: To review the basic concepts of CR and to provide a background for discussion of specific musculoskeletal applications of CR in children. MATERIALS AND METHODS: Various aspects of musculoskeletal CR in children are presented based on our 4 years' experience and a review of the literature. RESULTS: A greater amount of scatter capture occurs with storage phosphor CR than with a film-screen system in the 70- to 120-kVp range. This is attributed to a lower K-absorption edge of barium in the barium fluorohalide (BaFBr) compound used in the imaging plate. A significant reduction of scatter to primary radiation, improvement in bony trabecular sharpness, and improvement in line pair resolution can be achieved in pediatric musculoskeletal imaging using an air gap without an increase in the skin entrance dose as compared to the non-grid table top technique. With CR, in addition to proper radiographic exposure technique, one needs to preprogram and select the optimal processing technique for each anatomic region, projection and age group of the child. CONCLUSION: The main advantages of CR in pediatric musculoskeletal imaging consist of a reduction in radiation dose for many applications, improved contrast resolution, near elimination of repeat radiographs related to exposure errors, and digital processing capabilities for image enhancement, storage, retrieval, display and transmission. The current limitations of CR include the moderately high start-up cost, the long learning curve to produce optimal films, and the reduced spatial resolution. PMID- 9211948 TI - Detection of pulmonary metastases with pathological correlation: effect of breathing on the accuracy of spiral CT. AB - BACKGROUND: CT of the chest for suspected pulmonary metastases in adults is generally performed using a breath-hold technique. The results may not be applicable to young children in whom breath-holding may be impossible. OBJECTIVE: Determine the effect of breathing on the accuracy of pulmonary metastasis detection by spiral CT (SCT). MATERIALS AND METHODS: Prior to euthanasia four anesthetized dogs with metastatic osteosarcoma underwent SCT with a collimation of 5 mm and a pitch of 2, during both induced breath-hold and normal quiet breathing. Images were reconstructed as contiguous 5-mm slices. Macroscopically evident metastases were noted at postmortem. Hard-copy SCT images were reviewed by ten radiologists, each of whom circled all suspected metastases. SCT images were compared with postmortem results to determine true and false positives. RESULTS: The pathologist identified 132 macroscopically evident pulmonary metastases. For metastasis detection, there was no significant difference between breath-hold SCT and breathing SCT. CONCLUSION: In our animal model, SCT can be performed during normal resting breathing without significant loss of accuracy in the detection of pulmonary metastases. PMID- 9211949 TI - Alterations in spinal fluid drainage in infants with hydrocephalus. AB - We describe two cases of hydrocephalus in which spinal sonography revealed underlying causes responsible for the failure of therapeutic lumbar punctures. PMID- 9211950 TI - Cystic periventricular leukomalacia of the corpus callosum. AB - Periventricular leukomalacia (PVL) is a common finding during neurosonography of preterm infants. Secondary thinning of the corpus callosum is seen following PVL, typically from loss of hemispheric white matter tracts. We report a case of direct involvement of the corpus callosum with PVL, its pathogenesis, and its potential as a cause of corpus callosal thinning. PMID- 9211951 TI - An unusual foreign body: catfish spine. PMID- 9211952 TI - Isolated H-type recto-vaginal fistula associated with a vulval abscess. AB - Recto-vaginal fistula is well known to occur in association with imperforate anus. We describe the case of an isolated "H-type" recto-vaginal fistula with no other anorectal abnormalities. The patient presented at 2 months of age with a vulval abscess and passing faeces per vaginum. Unilateral renal agenesis was also seen in this patient. We are unaware of any previous reports in the English language literature of this isolated abnormality. PMID- 9211953 TI - Transjugular intrahepatic portosystemic shunt in an infant. AB - A 15-month-old girl, who presented with biliary cirrhosis secondary to cystic fibrosis with refractory ascites and recurrent intestinal bleeding, underwent percutaneous transjugular intrahepatic portosystemic shunting. Immediately following the procedure the ascites disappeared and no further bleeding occurred. The stent shunt was patent on Doppler ultrasound until the 22nd day. The patient died on day 22 because of liver failure due to a low-flow syndrome with severe hepatic ischaemia, but with no recurrence of bleeding or ascites. PMID- 9211954 TI - A horseshoe adrenal gland in an infant with asplenia. AB - Adrenal anomalies are rare, and when present are usually associated with renal malformations. In this article we present a case of "horseshoe" adrenal glands in a patient with asplenia, various cardiac anomalies and normal kidneys and bladder. PMID- 9211955 TI - Sonography of congenital adrenal hyperplasia due to partial deficiency of 3beta hydroxysteroid dehydrogenase: a case report. AB - We report the case of a patient with congenital adrenal hyperplasia (CAH) secondary to a partial deficiency of 3beta-hydroxysteroid dehydrogenase. This form occurs in less than 1 % of all patients with CAH. Sonographic evaluation of the adrenal glands demonstrated width and length measurements significantly above normal values. The sonographic findings are not diagnostic of the particular enzyme deficiency in CAH, and the exact etiology should be pursued with laboratory investigation. PMID- 9211956 TI - Idiopathic bilateral anterior mediastinal abscesses. AB - Mediastinal suppuration and abscess formation are uncommon in the era of antibiotics. The case of a child with seemingly idiopathic bilateral and separate anterior mediastinal abscesses is presented. The chest radiography, sonography and computed tomography findings are described. PMID- 9211957 TI - Evaluation of contrast media for bronchography. AB - BACKGROUND: Bronchography is occasionally needed for the evaluation and management of some congenital pulmonary anomalies as well as some acquired diseases, usually of the tracheo- bronchial tree. There is currently no effective, approved contrast agent for this imaging techniq ue. OBJECTIVE: We evaluated five agents (barium sulfate, iohexol, propyliodone oily, propyliodone aqueous, and perflubron) in terms of image quality, histologic changes, and effects on hemodynamics, blood gases, and standard laboratory tests in New Zealand White rabbits. MATERIALS AND METHODS: Animals were anesthetized and intubated. Each contrast agent (0.25 ml/kg) was administered intratracheally. Three animals in each group had intravenous lines placed for blood sampling and blood pressure monitoring and were sacrificed at 1 h. An additional three animals for each agent were sacrificed at 24 h and 1 week after imaging. Blood samples were taken immediately before contrast instillation and at 1 h postbronchography. Fluoroscopic images were recorded on standard VHS video tape and evaluated in blind fashion. Segments of lung tissue and bronchi were obtained for histologic examination. RESULTS: Necrosis and/or inflammatory infiltrates were noted in 78 % of the bronchograms performed with propyliodone aqueous, 67 % with propyliodone oily, 55 % with perflubron, and 33 % with iohexol 120, 240 and 350. No histologic damage was observed with barium. The propyliodones gave the best-quality imaging results and the most histologic changes. Iohexol, in any concentration, gave the least acceptable images and a moderate number of histologic changes. Barium sulfate demonstrated acceptable images with virtually no histologic changes. CONCLUSION: From the histologic and imaging results, barium is the best available contrast material for bronchography. PMID- 9211959 TI - Tandem balloon dilatation for childhood achalasia. AB - BACKGROUND: There are no previous reports of tandem balloon dilatation in childhood achalasia. OBJECTIVE: To report the treatment of four cases of paediatric achalasia using tandem balloon dilatation of the lower oesophageal sphincter. A review of the literature since 1986 was undertaken to compare outcomes of balloon dilatation and surgery. MATERIALS AND METHODS: A retrospective review of the patients diagnosed with this condition and treated at our institution over the past 6 years: all four patients were treated by balloon dilatation of the lower oesophageal sphincter using two or three balloons in tandem. The definition of technical success was demonstration of a waist at 1-1.5 atmospheres of inflation pressure followed by abolition of the waist at higher pressures. Where this was unable to be achieved using a single balloon, two or three balloons in tandem were used. RESULTS: No patient required oesophagomyotomy, and symptomatic control has been good to excellent in three of four patients. No significant side effects were encountered. CONCLUSIONS: Balloon dilatation and surgery have similar success rates in paediatric achalasia. Because of the low morbidity associated with balloon dilatation, the procedure should be considered as first line treatment of this condition. If the lower oesophageal sphincter is stretched insufficiently using a single balloon, tandem balloon dilatation should be utilised. PMID- 9211958 TI - The diagnosis of malrotation during air enema procedure. AB - This paper describes the correct diagnosis of cecal malposition, suggesting midgut malrotation, during air enema examination in seven patients. It is possible to diagnose cecal malposition by air enema, even in the presence of a reducible intussusception. PMID- 9211960 TI - Radiologic-Pathologic Conference of Children's Hospital Boston: bilateral asymptomatic renal enlargement. AB - An otherwise healthy 11-month-old boy presented with bilateral abdominal masses. Imaging findings, differential diagnosis, histological findings, and pertinent discussion are presented. PMID- 9211961 TI - Anastomosis of the umbilical arteries: physiologic variant or complication of umbilical catheterization? PMID- 9211962 TI - Enlarging giant liver cyst in Beckwith-Wiedemann syndrome. PMID- 9211963 TI - Peripheral primitive neuroectodermal tumor: report of a case arising in the kidney. PMID- 9211965 TI - Asen Hadjiolov, remembered. PMID- 9211966 TI - Eukaryotic ribosomal RNA: the recent excitement in the nucleotide modification problem. AB - Eukaryotic ribosomal RNA (rRNA) contains numerous modified nucleotides: about 115 methyl groups and some 95 pseudouridines in vertebrates; about 65 methyl groups and some 45 pseudouridines in Saccharomyces cerevisiae. All but about ten of the methyl groups are ribose methylations. The remaining ten are on heterocyclic bases. The ribose methylations occur very rapidly upon the primary rRNA transcript in the nucleolus, probably on nascent chains, and they appear to play an important role in ribosome maturation, at least in vertebrates. All of the methyl groups occur in the conserved core of rRNA. However, there is no consensus feature of sequence or secondary structure for the methylation sites; thus the nature of the signal(s) for site-specific methylations had until recently remained a mystery. The situation changed dramatically with the discovery that many of the ribose methylation sites are in regions that are precisely complementary to small nucleolar RNA (snoRNA) species. Experimental evidence indicates that structural motifs within the snoRNA species do indeed pinpoint the precise nucleotides to be methylated by the putative 2'-O-methyl transferase(s). Regarding base methylations, the gene DIM1, responsible for modification of the conserved dimethyladenosines near the 3' end of 18S rRNA, has been shown to be essential for viability in S. cerevisiae and is suggested to play a role in the nucleocytoplasmic transport of the small ribosomal subunit. Recently nearly all of the pseudouridines have also been mapped in the rRNA of several eukaryotic species. As is the case for ribose methylations, most pseudouridine modifications occur rapidly upon precursor rRNA, within core sequences, and in a variety of local primary and secondary structure environments. In contrast to ribose methylation, no potentially unifying process has yet been identified for the enzymic recognition of the many pseudouridine modification sites. However, the new data afford the basis for a search for any potential involvement of snoRNAs in the recognition process. PMID- 9211967 TI - U3 snoRNA may recycle through different compartments of the nucleolus. AB - A model is proposed in which U3 small nucleolar RNA (snoRNA) is recruited from an inactive, stored form in the dense fibrillar component (DFC) of the nucleolus to an active form that is associated with the initial ribosomal RNA (rRNA) precursor. The initial steps of rRNA processing occur in the DFC, and then it is proposed that the U3 snoRNA moves with intermediates in rRNA processing from the DFC to the granular component (GC) of the nucleolus. The nucleolar protein fibrillarin is located primarily in the DFC, and it is suggested that the complex of fibrillarin and U3 snoRNA dissociates when U3 snoRNA transits to the GC. Finally, when U3 snoRNA is released from the processed rRNA, the tether holding the rRNA in the nucleolus is broken and rRNA can then be exported from the nucleolus to the cytoplasm. U3 snoRNA is hypothesized to recycle back from the GC to the DFC where it is stored until future association with another initial rRNA precursor. Data supporting this model are summarized. U3 snoRNA is also stored in the coiled body of interphase cells and in the nucleolar remnants and prenucleolar bodies of mitotic cells, and there may be some similarity in the binding sites for stored U3 snoRNA in the DFC and in these structures. PMID- 9211968 TI - A class of nonribosomal nucleolar components is located in chromosome periphery and in nucleolus-derived foci during anaphase and telophase. AB - . The subcellular location of several nonribosomal nucleolar proteins was examined at various stages of mitosis in synchronized mammalian cell lines including HeLa, 3T3, COS-7 and HIV-1 Rev-expressing CMT3 cells. Nucleolar proteins B23, fibrillarin, nucleolin and p52 as well as U3 snoRNA were located partially in the peripheral regions of chromosomes from prometaphase to early telophase. However, these proteins were also found in large cytoplasmic particles, 1-2 microm in diameter, termed nucleolus-derived foci (NDF). The NDF reached maximum numbers (as many as 100 per cell) during mid- to late anaphase, after which their number declined to a few or none during late telophase. The decline in the number of NDF approximately coincided with the appearance of prenucleolar bodies and reforming nucleoli. The HIV-1 Rev protein and a mutant Rev protein defective in its nuclear export signal were also found in the NDF. The mutant Rev protein precisely followed the pattern of localization of the above nucleolar proteins, whereas the wild-type Rev did not enter nuclei until G1 phase. The nucleolar shuttling phosphoprotein Nopp140 did not follow the above pattern of localization during mitosis: it dispersed in the cytoplasm from prometaphase through early telophase and was not found in the NDF. Although the NDF and mitotic coiled bodies disappeared from the cytoplasm at approximately the same time during mitosis, protein B23 was not found in mitotic coiled bodies, nor was p80 coilin present in the NDF. These results suggest that a class of proteins involved in preribosomal RNA processing associate with chromosome periphery and with NDF as part of a system to conserve and deliver preexisting components to reforming nucleoli during mitosis. PMID- 9211970 TI - Beyond ribosomal DNA: on towards the telomere. AB - We have sequenced and analyzed 8.3 kb of sequence adjacent and distal to the human ribosomal DNA (rDNA); this distal sequence connects to the rDNA cluster just 4 kb upstream of the first promoter and is shared among the acrocentric chromosomes and, at least in part, it is also present in other primates. The sequence differs in character from that of the rDNA intergenic spacer (IGS) in that it does not contain long stretches of either polypyrimidine or polypurine. However, just like the IGS, it contains numerous repetitive elements, including retroposed fragments of 28S rRNA and large pieces of the IGS. In addition, we show that the rDNA clusters are not interrupted by other sequences and do not recombine with this distal segment. PMID- 9211969 TI - Experimental induction of prenucleolar bodies (PNBs) in interphase cells: interphase PNBs show similar characteristics as those typically observed at telophase of mitosis in untreated cells. AB - Recently, it was shown that a short exposure of living mammalian cells to low ionic strength buffers (hypotonic shock) caused partial or almost complete unraveling of interphase nucleoli. However, when the cells were released from the hypotonic shock and transferred to normal isotonic medium, functionally active and structurally integral nucleoli were reassembled at their initial positions within interphase nuclei. Here, we show further that this process is accompanied by the appearance of numerous discrete extranucleolar bodies, which have striking similarities to the prenucleolar bodies (PNBs) observed in untreated cells at telophase of mitosis. (1) Like PNBs at mitosis, hypotonically induced interphase PNBs are composed of RNA-positive granules and fibrils, contain the major nucleolar protein B23 and silver-binding proteins, but lack DNA and RNA polymerase I transcription factor UBF. (2) As for mitotic PNBs, disappearance of the interphase PNB counterparts coincides with the increase in size of reconstructed nucleoli. (3) Addition of actinomycin D does not prevent assembly of interphase PNBs, but does arrest their coalescence with the chromosomal nucleolus-organizing regions and blocks the complete reformation of nucleoli. It is concluded that the assembly of PNBs generally observed at telophase of mitosis can be induced experimentally in nuclei of interphase mammalian cells in vivo. At interphase, this process is probably initiated by changes in the intracellular ionic environment. PMID- 9211971 TI - "Micronucleoli" in the Xenopus germinal vesicle. AB - The germinal vesicle of the Xenopus oocyte contains 1500 or more extrachromosomal nucleoli that are assembled on amplified copies of the rRNA genes. Many of these nucleoli have diameters of 10-15 micron, but some are much smaller, ranging down to 1 micron or less. Morphologically the smaller nucleoli or "micronucleoli" resemble the similarly sized B snurposomes, but they can be recognized with appropriate antibody probes (e.g., anti-nucleolin and anti-fibrillarin). We describe here a sensitive fluorescent staining technique that uses avidin and propidium iodide to visualize the rDNA in the amplified nucleoli. Many large nucleoli stain about as brightly as haploid yeast nuclei on the same slides. They presumably contain about 12 Mb of DNA, equivalent to 900 rDNA repeats. The smallest micronucleoli display only a tiny dot of stain, which must correspond to relatively few rDNA repeats. PMID- 9211972 TI - Does Saccharomyces need an organized nucleolus? AB - The ribosomal RNA (rRNA) genes of most eukaryotic organisms are arranged in one or more tandem arrays, often termed nucleolar organizer regions. The biological implications of this tandem organization are not known. We have tested the requirement for such a chromosomal organization by directly comparing the transcription and processing of rRNA in isogenic strains of Saccharomyces cerevisiae that differ only in the organization of their rRNA genes. Strain L 1489 carries the RDN locus, consisting of 100-150 copies of the rRNA genes in a tandem array on chromosome XII. Strain L-1521 has a complete deletion of the RDN array, but carries many copies of a plasmid that includes a single rRNA gene. While this strain grows reasonably well, the average transcriptional activity of the plasmid genes is substantially less than that of the chromosomal copies. However, there is little difference in the processing of the 35S pre-rRNA to the mature 25S:5.8S and 18S products. Thus, neither a chromosomal location nor a tandem repeat of the rRNA genes is important for the assembly and function of the many protein and RNA molecules necessary for the processing of the rRNA transcripts. We investigated the consequence of a dispersed gene arrangement on nucleolar structure. Immunofluorescence microscopy revealed that in strain L-1521 the yeast fibrillarin, Nop1p, rather than being confined to a defined nucleolus at the edge of the nucleus as it is in cells with the normal arrangement of rRNA genes, is spread throughout the nucleus. This observation implies that each plasmid rRNA gene can serve as a nucleolar organizer. Finally, data from pulse labeling experiments show that the repression of rRNA transcription due to failure of the secretory pathway is independent of whether the rRNA genes are at the RDN locus on chromosome XII or on plasmids. This result suggests that the regulation of rRNA transcription occurs at the level of soluble factors rather than chromatin structure. PMID- 9211973 TI - Small nucleolar RNAs and nucleolar proteins in Xenopus anucleolate embryos. AB - We investigated the presence and localization, in the cells of anucleolate mutant embryos of Xenopus laevis, of three representative small nucleolar RNAs (snoRNAs), U3, U15 and U17, and of two nucleolar proteins, nucleolin and fibrillarin. The levels of the three snoRNAs in the anucleolate mutant are the same as in normal embryos, in contrast to 5S RNA and ribosomal proteins. In situ hybridization showed that, in the absence of fully organized nucleoli, the three RNAs are diffusely distributed in the nucleus and partly associated with a number of small structures. Nucleolin and fibrillarin are also present in the anucleolate embryos as in normal embryos, although there is less nucleolin mRNA in the former. The two nucleolar proteins were localized by immunofluorescence microscopy. Fibrillarin, similar to its associated U3 and U15 snoRNAs, is diffusely distributed in the anucleolate nucleus and is partly associated with small structures, probably prenucleolar bodies and pseudonucleoli. Nucleolin also appears diffusely distributed in the nucleus with some spots of higher concentration, but with a different pattern with respect to fibrillarin. PMID- 9211974 TI - Relative distribution of rDNA and proteins of the RNA polymerase I transcription machinery at chromosomal NORs. AB - Using confocal and immunofluorescence microscopy the relative distribution of the ribosomal chromatin and some proteins of the RNA polymerase I transcription machinery such as upstream binding factor (UBF), RNA polymerase I and DNA topoisomerase I was analyzed on chromosomal nucleolus organizer regions (NORs) of PtK1 cells. Staining with various DNA fluorochromes revealed that the ribosomal chromatin may be found at the axial region of the NOR and also at lateral expansions around the axis that can also be detected by in situ hybridization. It was observed that the transcription machinery shows a crescent-shaped distribution around the axial ribosomal chromatin at the NOR of metaphase and anaphase chromatids. An ultrastructural analysis of serially sectioned NORs supports this crescent-shape organization. Taking into account previous and present results and the loop/scaffold model of chromosome structure, we propose a model of NOR organization. The model proposes that ribosomal genes that were inactive in the preceding interphase would be present as condensed short Q-loops occupying the axial region of the NOR. Ribosomal genes previously active during interphase would be undercondensed as large R-loops associated with the transcription machinery, which is distributed in a crescent-shaped fashion around the previously active ribosomal DNA. PMID- 9211975 TI - Looking at Christmas trees in the nucleolus. AB - We describe novel nucleolar structures, observed by thin section electron microscopy in oocyte nuclei of the grasshopper Locusta migratoria, which we interpret, based on morphological and compositional criteria, as rDNA transcription units. Morphologically they resemble the condensed and foreshortened "Christmas trees" seen in Miller spreads of nucleolar chromatin prepared from the same biological material. They contain DNA and rRNA as shown by immunocytochemistry and in situ hybridization and are concentrated in several intranucleolar cavities. The presumptive rDNA transcription units extend throughout the interior of these nucleolar pockets or are selectively enriched at their outermost zones in close contact with the surrounding fibrillarin-positive dense component. We suggest that the nucleolar pockets of Locusta oocytes are equivalent to the fibrillar centers of somatic nucleoli and discuss possible implications for the current understanding of the functional organization of nucleoli. PMID- 9211976 TI - Analysis of nucleolar transcription and processing domains and pre-rRNA movements by in situ hybridization. AB - We have examined the cytological localization of rRNA synthesis, transport, and processing events within the mammalian cell nucleolus by double-label fluorescent in situ hybridization analysis using probes for small selected segments of pre rRNA, which have known half-lives. In particular, a probe for an extremely short lived 5' region that is not found separate of the pre-rRNA identifies nascent transcripts within the nucleolus of an intact active cell, while other characterized probes identify molecules at different stages in the rRNA processing pathway. Through these studies, visualized by confocal and normal light microscopy, we (1) confirm that rDNA transcription occurs in small foci within nucleoli, (2) show that the nascent pre-rRNA transcripts and most likely also the rDNA templates are surprisingly extended in the nucleolus, (3) provide evidence that the 5' end of the nascent rRNA transcript moves more rapidly away from the template DNA than does the 3' end of the newly released transcript, and (4) demonstrate that the various subsequent rRNA processing steps occur sequentially further from the transcription site, with each early processing event taking place in a distinct nucleolar subdomain. These last three points are contrary to the generally accepted paradigms of nucleolar organization and function. Our findings also imply that the nucleolus is considerably more complex than the conventional view, inferred from electron micrographs, of only three kinds of regions - fibrillar centers, dense fibrillar components, and granular components - for the dense fibrillar component evidently consists of several functionally distinct sub-domains that correlate with different steps of ribosome biogenesis. PMID- 9211977 TI - Early stages of pre-rRNA formation within the nucleolar ultrastructure of mouse cells studied by in situ hybridization with a 5'ETS leader probe. AB - The first cleavage in the processing of the rRNA primary transcript in mammals occurs within the 5'-terminal region of the 5' external transcribed spacer (5'ETS), which makes the upstream portion of this spacer a selective marker of nascent transcripts. Moreover, short treatments with low doses of actinomycin D (AMD), which selectively suppress pre-rRNA synthesis and allow processing of preformed pre-rRNAs, result in the production of prematurely terminated transcripts essentially spanning the 5'ETS leader region. To gain further insight into the intranucleolar localization of early stages of preribosome formation we analyzed the distribution of this specific pre-rRNA segment by in situ hybridization at the ultrastructural level in AMD-treated or in control 3T3 mouse cells. In control cells, 5'ETS leader rRNA was detected at the border of the fibrillar centers and over the dense fibrillar component, in agreement with previous data suggesting that rRNA gene transcription takes place at the border of the fibrillar centers before a rapid transfer of the nascent trancript to the dense fibrillar component. Observation of cells subjected to a short treatment with low doses of AMD fully supports this conclusion, with the prematurely terminated 5'ETS leader-containing transcripts detected at the border of enlarged fibrillar centers. With prolonged periods of AMD treatment even the partial transcription of rRNA genes is blocked and fibrillar centers of typically segregated nucleoli show no positive signals with the 5'ETS leader probe. We also analyzed in parallel the intranucleolar distribution of U3 small nucleolar RNA, which is involved in 5'ETS processing, by hybridization with biotinylated antisense oligonucleotides. Distribution of U3 roughly paralleled that of 5'ETS leader rRNA in untreated cells. However, U3 RNA persisted in the dense fibrillar component of segregated nucleoli whatever the conditions of drug treatment, i.e., even after a thorough chase of the rRNA precursors from this nucleolar compartment. PMID- 9211979 TI - U14 small nucleolar RNA makes multiple contacts with the pre-ribosomal RNA. AB - The small nucleolar RNA (snoRNA) U14 has two regions of extended primary sequence complementarity to the 18S rRNA. The 3' region (domain B) shows the consensus structure for the methylation guide class of snoRNAs, whereas base-pairing between the 5' region (domain A) and the 18S rRNA sequence is required for the formation of functional ribosomes. Between domains A and B lies another essential region (domain Y). Here we report that yeast U14 can be cross-linked in vivo to the pre-rRNA; cross-linking is detected exclusively with the 35S primary transcript. Many nucleotides in U14 that lie outside of domains A and B are cross linked to the pre-rRNA; in particular the essential domain Y region is cross linked at several sites. U14 is, therefore, in far more extensive contact with the pre-rRNA than predicted from simple base-pairing models. Moreover, U14 can be cross-linked to other small RNA species. The functional interactions made by U14 during ribosome synthesis are likely to be very complex. PMID- 9211978 TI - Delocalization of some small nucleolar RNPs after actinomycin D treatment to deplete early pre-rRNAs. AB - Retention of some components within the nucleolus correlates with the presence of rRNA precursors found early in the rRNA processing pathway. Specifically, after most 40S, 38S and 36S pre-rRNAs have been depleted by incubation of Xenopus kidney cells in 0.05 microg/ml actinomycin D for 4 h, only 69% U3 small nucleolar RNA (snoRNA), 68% U14 snoRNA and 72% fibrillarin are retained in the nucleolus as compared with control cells. These nucleolar components are important for processing steps in the pathway that gives rise to 18S rRNA. In contrast, U8 snoRNA, which is used for 5.8S and 28S rRNA production, is fully retained in the nucleolus after actinomycin D treatment. Therefore, U8 snoRNA is in a different category than U3 and U14 snoRNA and fibrillarin. It is proposed that U3 and U14 snoRNA and fibrillarin, but not U8 snoRNA, bind to the external transcribed spacer or internal transcribed spacer 1, and when these binding sites are lost after actinomycin D treatment some of these components cannot be retained in the nucleolus. Other binding sites may also exist, which would explain why only some and not all of these components are lost from the nucleolus. PMID- 9211980 TI - Variable region V1 of Saccharomyces cerevisiae 18S rRNA participates in biogenesis and function of the small ribosomal subunit. AB - The role of helix 6, which forms the major portion of the most 5'-located expansion segment of Saccharomyces cerevisiae 18S rRNA, was studied by in vivo mutational analysis. Mutations that increased the size of the helical part and/or the loop, even to a relatively small extent, abolished 18S rRNA formation almost completely. Concomitantly, 35S pre-rRNA and an abnormal 23S precursor species accumulated. rDNA units containing these mutations did not support cell growth. A deletion removing helix 6 almost completely, on the other hand, had a much less severe effect on the formation of 18S rRNA, and cells expressing only the mutant rRNA remained able to grow, albeit at a much reduced rate. Disruption of the apical A.U base pair by a single point mutation did not cause a noticeable reduction in the level of 18S rRNA but did result in a twofold lower growth rate of the cells. This effect could not be reversed by introduction of a second point mutation that restores base pairing. We conclude that both the primary and the secondary structure of helix 6 play an important role in the formation and the biological function of the 40S subunit. PMID- 9211981 TI - Mitosis-specific phosphorylation of gar2, a fission yeast nucleolar protein structurally related to nucleolin. AB - The nucleolar protein gar2 of fission yeast is structurally related to the multifunctional nucleolar protein nucleolin from vertebrates and has been shown to be implicated in production of 18S rRNA. gar2 contains several potential casein kinase 2 (CK2) phosphorylation sites and a single putative p34(cdc2 )phosphorylation site in the consensus S50PKK. Here, we show that, like nucleolin, gar2 is phosphorylated in vitro by both highly purified CK2 from CHO cells and p34(cdc2 )from starfish oocytes. Moreover, the substitution of alanine for the N-terminal serine 50 abolishes phosphorylation by p34(cdc2 )in vitro. We also provide evidence that gar2 is phosphorylated in vitro by a p13(suc1) Sepharose-bound kinase from Schizosaccharomyces pombe extracts that displays cell cycle-regulated activity similar to that of the p34(cdc2(kinase. In vivo 32P labeling of cells indicates that gar2 is a phosphoprotein and that incorporation of phosphate on residue 50 occurs specifically at mitosis. Taken together, these results lead us to propose that gar2 is likely to be an in vivo substrate for the mitotic p34(cdc2 )kinase. However, this posttranslational modification of the gar2 protein does not appear to be essential for normal production of 18S rRNA. PMID- 9211983 TI - Human proliferation-related protein p120 interacts with HSRP1. AB - Protein p120 is a proliferation-related nucleolar protein, which is detectable early in the G1-phase of the cell cycle and peaks early in the S-phase. Most human malignant tumor cells contain much higher levels of protein p120 than do normal resting cells. To learn more about p120-associated proteins, a yeast two hybrid screen was carried out using protein p120 as the bait. Five positive clones were identified from 2 million clones for further analysis. Three of them encoded portions of the same protein, which had identity to human SRP1. The recombinant p120 and HSRP1 proteins produced in Sf9 cells are associated with each other, confirming the results of the yeast genetic assay. Protein SRP1 was originally characterized as a suppressor of RNA polymerase I mutations, and recently human SRP1 was identified as a receptor for nuclear localization sequences. In the present study, use of deletion mutations revealed that the binding of human SRP1 to p120 required the p120 nuclear localization signal (amino acids 96-119) and the C-terminus of human SRP1 (amino acids 453-491). PMID- 9211982 TI - Ultrastructural changes in the Schizosaccharomyces pombe nucleolus following the disruption of the gar2+ gene, which encodes a nucleolar protein structurally related to nucleolin. AB - The nucleolar protein gar2, from the fission yeast Schizosaccharomyces pombe, is the functional homolog of NSR1 from Saccharomyces cerevisiae, and is structurally related to nucleolin from vertebrates. By immunocytochemistry at the electron microscope level, we show that gar2 co-localizes with RNA polymerase I and the gar1 protein along the dense fibrillar component of the nucleolus in a wild-type strain of S. pombe, suggesting that gar2 is involved in the transcription and/or in the early steps of maturation of the ribosomal RNAs. Since the effects of disruption of the gar2+ gene might also shed light on the role of the gar2 protein, we analyzed the ultrastructure of the nucleolus of a gar2-disruption mutant. The nucleolus of the gar2- mutant is dramatically reorganized when compared with that of the wild-type gar2+ strain: a truncated protein containing the NH2-terminus of the gar2 protein is accumulated in an unusual nucleolar "dense body". Our results also suggest that the NH2-terminus might be sufficient for nucleolar localization via interaction with specific nucleolar components and support the hypothesis that gar2 in wild-type S. pombe interacts with nascent pre rRNA via its two RNA-binding domains in combination with the glycine/arginine rich domain. We also report that disruption of the gar2+ gene results in a mutant that is defective in cytokinesis and nuclear division. PMID- 9211984 TI - Neural cell adhesion molecule L1: signaling pathways and growth cone motility. AB - The neural cell adhesion molecule L1 plays a key role in nervous system development including neuronal migration, neurite growth, and axonal fasciculation. L1 is expressed on most developing axons, and homophilic binding of L1 molecules on adjacent axons is likely to play a key role in axon extension. It is now well documented that a number of second-messenger systems are involved in L1-stimulated neurite growth in vitro. However, it is unclear how L1 homophilic or heterophilic binding trigger signals that regulate the mechanical forces that produce axon extension. In this report, we will review recent advances in understanding L1-associated signals, L1 interactions with the cytoskeleton, and the molecular mechanisms underlying growth cone motility. PMID- 9211985 TI - Association of human, rat, and rabbit apolipoprotein E with beta-amyloid. AB - In humans, apolipoprotein E (apoE) has three major isoforms, E2 (Cys112, Cys158), E3 (Cys112, Arg158), and E4 (Arg112, Arg158). While epsilon4 is a genetic risk factor for Alzheimer's disease (AD), epsilon2 may protect against late-onset AD. Using native preparations of apoE from conditioned tissue culture media or plasma lipoproteins, we have previously shown that when equivalent amounts of apoE3 or E4 were incubated with beta-amyloid (A beta), apoE3 formed 20 times as much SDS stable complex with the peptide as apoE4. This preferential binding of A beta to apoE3 was abolished when apoE was purified by a process which includes delipidation and denaturation. Here we expand these observations to include A beta binding to lipoprotein-associated and purified apoE2. Lipoproteins isolated from the plasma of individuals homozygous for either epsilon2 or epsilon3 were incubated with A beta(1-40). SDS-stable complex formation was analyzed by a non reducing gel shift assay, followed by immunoblotting with either A beta or apoE antibodies. ApoE2:A beta complex formation was comparable to apoE3:A beta in both native and purified preparations of apoE. In addition, lipoprotein-associated rat apoE (Arg112, Arg158), like human apoE4, did not form complex with A beta, while lipoprotein-associated rabbit apoE (Cys112, Arg158) did bind the peptide. These binding studies provide one possible explanation for protective effects of both apoE2 and E3 against the development of Alzheimer's disease. PMID- 9211986 TI - Reduction of connexin43 expression and dye-coupling during neuronal differentiation of human NTera2/clone D1 cells. AB - Gap junctions are plasma membrane specializations that allow direct communication among adjoining cells. We used a human pluripotential teratocarcinoma cell line, NTera-2/clone D1 (NT2/D1), as a model to study gap junctions in CNS neurons and their neuronal precursors. These cells were differentiated following retinoic acid (RA) treatment for 4 weeks and antiproliferative agents for 3 weeks, respectively, to yield post-mitotic CNS neuronal (NT2-N) cells. The cytoplasmic RNA was isolated from NT2/D1 cells both before and during RA treatment and from differentiated neurons (NT2-N cells). These RNA samples were examined using Northern blot analysis with cDNA probes specific for connexin26, -32, and -43. Connexin26 and -32 mRNAs were absent in NT2/D1 and NT2-N cells. Connexin43 mRNA was expressed at high levels in NT2/D1 cells before RA treatment, but it decreased significantly during RA induction. There was no detectable connexin43 mRNA in NT2-N cells. Western blot analysis confirmed the expression of connexin43 protein in NT2/D1 cells before and during RA treatment. The protein profile detected in Western blot analysis indicated two bands representing different phosphorylation states of connexin43. Our immunocytochemistry results did not show connexin26 and -32 immunoreactivity in NT2/D1 and NT2-N cells. However, we detected connexin43 immunoreactivity in NT2/D1 cells with a decreasing pattern upon RA induction. Both Western blotting and immunocytochemistry confirmed the absence of connexin43 protein in NT2-N cells. NT2/D1 cells passed calcein readily to an average of 18 cells, confirming the functionality of gap junctions in these cells. The extent of dye-coupling decreased about 78% when NT2/D1 cells were RA treated for 4 weeks. NT2-N differentiated neurons did not pass dye to the adjacent cells. We conclude that both connexin43 expression and dye coupling capacity decrease during neuronal differentiation of NT2/D1 cells. PMID- 9211987 TI - Identification of transcriptionally regulated mRNAs from mouse Schwann cell precursors using modified RNA fingerprinting methods. AB - We have adopted RNA fingerprinting methods to screen for genes that are rapidly up- or down-regulated during normal mammalian development, comparing mRNA from early (embryo day 12) to late (embryo day 13) mouse Schwann cell precursors. The use of total RNA, a reduction of cDNA template for amplification, the detection of RT-PCR products with a sensitive DNA stain and polyacrylamide gel electrophoresis and rigid selection criteria involving three screening steps are significant improvements on previous methods. Of 19 differentially displayed bands, 15 represented novel genes. The four known cDNA fragments (interleukin enhancer binding factor 1, beta3 subunit of phospholipase C, brain beta-spectrin, and P21 polypeptide) consisted of coding sequences, indicating a high chance of obtaining coding regions. A semiquantitative RT-PCR analysis of three of the four known genes and a cDNA fragment randomly selected from the pool of 15 novel sequences, confirmed that they were regulated between embryo days 12 and 13, as predicted by the display gels. Our results suggest that the combination of methods described here will have wide applicability in studies of other developmental systems where precisely timed changes occur and where only small amounts of RNA can be obtained for analysis. PMID- 9211988 TI - Identification and characterization of P2Y2 nucleotide receptors in human retinal pigment epithelial cells. AB - P2 nucleotide receptor expression in cultured human retinal pigment epithelial (RPE) cells was investigated using the photoaffinity ATP analog BzATP, polymerase chain reaction of reverse-transcribed RNA (RT-PCR) and fura-2 fluorescence measurement of changes in intracellular free calcium concentration ([Ca2+]i). In experiments carried out in RPE cells at passage 10-15, addition of micromolar concentrations of ATP, UTP, and ATPgammaS to RPE cells resulted in a rapid, transient 3.5-fold increase in [Ca2+]i followed by a prolonged elevation that was twofold above the original baseline. Similar results were obtained from cells at passage 2. Characteristics of nucleotide-stimulated calcium mobilization in RPE cells, including partial inhibition by pertussis toxin, suggest that a G protein coupled receptor mediates this response. Consistent with the expression of a P2Y2 nucleotide receptor subtype in RPE cells, [alpha-32P]BzATP labeled a 53-kDa protein in plasma membranes, and RT-PCR revealed the presence of P2Y2 receptor RNA. Adenosine had no effect on [Ca2+]i in RPE cells, indicating that the A2 subtype of P1 receptor described previously in human RPE is not involved in the response to nucleotides. Together the results indicate that human RPE cells express functional P2Y2 nucleotide receptors. PMID- 9211990 TI - Attempts to produce astrocyte cultures devoid of oligodendrocyte generating potential by the use of antimitotic treatment reveal the presence of quiescent oligodendrocyte precursors. AB - The presence of oligodendrocyte precursor cells which cannot be removed from primary cultures by antibody-dependent techniques complicates the interpretation of transplantation experiments designed to examine the potential of astrocytes to influence remyelination (Blakemore et al.; Glia 13:79-91, 1995). In the present series of experiments we have investigated the use of the antimitotic cytosine arabinoside to eliminate oligodendrocyte precursors from mixed glial cell cultures following immunolytic removal of both oligodendrocytes and oligodendrocyte progenitors using A2B5 and O4 monoclonal antibodies. Our results indicate that not all oligodendrocyte precursors are involved during the subsequent regeneration of oligodendrocytes since a population of precursors survive 3-day and 12-day exposure to cytosine arabinoside. Maintaining immunolysed cultures in serum-free medium containing cytosine arabinoside for 23 days, removed the potential to generate large clones of oligodendrocytes both in vitro and following transplantation. However, a small number of oligodendrocyte precursors survived this treatment and generated single oligodendrocytes in vitro and isolated clusters of oligodendrocyte remyelination following transplantation. Overall, these results indicate that oligodendrocyte precursors have considerable potential to generate oligodendrocytes, but, since they can also survive for long periods in a quiescent state, their complete elimination from immunolysed astrocyte cultures by the use of an antimitotic is unreliable, if not impossible. PMID- 9211989 TI - Neuronal differentiation in SH-SY5Y human neuroblastoma cells induces synthesis and secretion of tenascin and upregulation of alpha(v) integrin receptors. AB - Human SH-SY5Y neuroblastoma cells were induced to neuronal differentiation by using 12-0-tetradecanoylphorbol-13-acetate (TPA) and retinoic acid (RA). Both treatments rapidly induced long neurites and increased the content of neurofilaments as shown by immunocytochemistry and immunoblotting. Immunoprecipitation and immunoblotting of the culture medium with monoclonal antibodies demonstrated a rapid onset of synthesis and secretion of Mr 280,000 tenascin (Tn) polypeptide with TPA and both Mr 280,000 and 190,000 Tn polypeptides with RA and an increased secretion of extradomain A cellular fibronectin (EDA-Fn) upon both treatments. Upon RA treatment both Tn polypeptides were also found in extracellular matrix preparations of the differentiated cells. A diffuse extracellular Tn immunoreactivity and a distinct cytoplasmic reaction were seen in differentiated cells especially after exposure to monensin to inhibit cellular secretion. Instead, immunoprecipitation experiments suggested that laminin was synthesized by the cells but was not upregulated upon differentiation. Experiments with purified Tn, used to coat the culture substratum, demonstrated that the undifferentiated cells were unable to adhere or spread on Tn but rapidly acquired the spreading capacity upon differentiation with the inducing agents. In immunofluorescence and immunoblotting the undifferentiated cells presented only a faint heterogenous reaction for beta1 integrin (Int) subunit, whereas cells exposed to RA presented a strong reaction for the Int alpha1 and beta1 subunits, hence suggestive of Int alpha1beta1, and for Int alpha(v) subunit. Cells exposed to TPA showed an enhanced immunoreaction for Int alpha2 and beta1 subunits, suggestive of Int alpha2beta1, and for Int alpha(v) subunit. Immunoreactivity for Int alpha(v) located to distinct punctate plaques in the differentiated cells after both inducing agents. The results suggest that Tn is produced by cultured neuronally differentiating cells, and it is accompanied by the acquitance of an adhesion receptor for Tn. PMID- 9211992 TI - Role of P-170 glycoprotein in colchicine brain uptake. AB - To study the role of P-glycoprotein (P-gp) in the delivery of colchicine from blood to brain, the pharmacokinetics of colchicine in plasma and brain was studied in the rat by an in vivo method and by the in situ brain perfusion technique. Colchicine was administered intravenously at three doses (1, 2.5, and 5 mg/kg) with or without an inhibitor of P-gp, verapamil (0.5 mg/kg i.v.); blood and brain samples were taken at t = 1, 2, and 3 hr. Areas under the colchicine curve at doses from 2.5 to 5 mg/kg were proportional to dose for plasma but not for brain. At a colchicine dose of 5 mg/kg, verapamil co-treated rats showed a 1.65-fold enhancement of the colchicine concentration in plasma but a 4.5-fold enhancement in brain. During short experimental times (in situ brain perfusion technique), a comparable enhancement was found (4.26-fold): mean distribution volumes of colchicine were enhanced from 0.23 +/- 0.17 to 0.98 +/- 0.19 microl/g for the eight gray areas, and no effect was observed in the choroid plexus, which do not express P-gp. These results clearly show that P-gp, present at the luminal surface of the capillary endothelial cells, is responsible for the weak penetration of colchicine into the brain. PMID- 9211991 TI - Ruthenium red neurotoxicity and interaction with gangliosides in primary cortical cultures. AB - Ruthenium red (RR) is an inorganic polycationic dye able to exert several effects on the nervous system, including neurodegeneration, both in vivo and in cell cultures. Gangliosides have been shown to protect cultured neurons against several damaging conditions, and it has been postulated that RR can interact with the negative charges of the sialic acid residues of these molecules. In the present work we have tested the effect of the trisialoganglioside GT1b and the monosialoganglioside GM1 on the RR-induced neuronal damage in primary cortical cultures, as well as on the binding of RR to synaptosomes. GT1b at 100-200 microM concentrations partially protected against RR-induced neurodegeneration, as judged by light microscopy and by measurement of the reduction of a tetrazolium salt, while GM1 was ineffective. GT1b, but not GM1, also partly blocked both RR binding and its diminution in the culture medium occurring during incubation. These results suggest that the three negative charges of GT1b enable it to interact with RR and as a consequence the entrance of the dye into the cells is blocked and neurotoxicity is diminished, although other mechanisms of protection cannot be excluded. Endogenous polysialic acid-containing molecules do not seem to be involved in RR effects, since the removal of sialic acid residues by treatment with neuraminidase did not prevent the cell damage. PMID- 9211993 TI - Regulation of cerebellar nitric oxide production in response to prolonged in vivo hypoxia. AB - The aim of this study was to assess the influence of prolonged in vivo hypoxia on cerebellar nitric oxide (NO) production. Conscious rats were exposed to 10% O2 (balanced N2) for 12 or 48 hr (arterial PO2 between 35 and 39 mmHg). The animals were then killed, and the cerebella were quickly frozen. NO production was measured in vitro by monitoring the conversion of L-[3H]arginine to L [3H]citrulline. Protein and mRNA expressions of the neuronal (nNOS) and endothelial (ecNOS) isoforms of nitric oxide synthases were assessed by using immunoblotting and semiquantitative reverse transcription-polymerase chain reaction, respectively. We also measured mRNA expression of GTP cyclohydrolase I, the rate-limiting enzyme in the synthesis of NOS cofactor, tetrahydrobiopterin, and mRNA and protein expressions of argininosuccinate synthase and argininosuccinate lyase, essential enzymes for the recycling of L-citrulline to L arginine. Prolonged in vivo hypoxia resulted in a time-dependent increase in cerebellar nitric oxide synthase activity and a significant rise in mRNA and protein expressions of nNOS isoform; however, ecNOS protein expression declined significantly. There was also a rise in mRNA expression of GT cyclohydrolase I; however, neither mRNA nor protein expression of argininosuccinate synthase and argininosuccinate lyase changed significantly in hypoxic animals. These results suggest that prolonged hypoxia increases cerebellar NO formation as a result of upregulation of cerebellar nNOS expression, whereas ecNOS expression declines. We propose that cofactor availability for NO production may also increase during hypoxia because of upregulation of GTP cyclohydrolase I expression. Recycling of L-citrulline to L-arginine, however, remains unchanged. PMID- 9211994 TI - Changes in glial K+ currents with decreased extracellular volume in developing rat white matter. AB - Whole cell patch-clamp recordings of K+ currents from oligodendrocyte precursors in 10-day-old rats (P10) and, following myelination, in mature oligodendrocytes from 20-day-old rats (P20) were correlated with extracellular space (ECS) diffusion parameters measured by the local diffusion of iontophoretically injected tetramethylammonium ions (TMA+). The aim of this study was to find an explanation for the changes in glial currents that occur with myelination. Oligodendrocyte precursors (P10) in slices from corpus callosum were characterized by the presence of A-type K+ currents, delayed and inward rectifier currents, and lack of tail currents after the offset of a voltage jump. Mature oligodendrocytes in corpus callosum slices from P20 rats were characterized by passive, decaying currents and large tail currents after the offset of a voltage jump. Measurements of the reversal potential for the tail currents indicate that they result from increases in [K+]e by an average of 32 mM during a 20 msec 100 mV voltage step. Concomitant with the change in oligodendrocyte electrophysiological behavior after myelination there is a decrease in the ECS of the corpus callosum. ECS volume decreases from 36% (P9-10) to 25% (P20-21) of total tissue volume. ECS tortuosity lambda = (D/ADC)0.5, where D is the free diffusion coefficient and ADC is the apparent diffusion coefficient of TMA+ in the brain, increases as measured perpendicular to the axons from 1.53 +/- 0.02 (n = 6, mean +/- SEM) to 1.70 +/- 0.02 (n = 6). TMA+ non-specific uptake (k') was significantly larger at P20 (5.2 +/- 0.6 x 10(-3) s(-1), n = 6) than at P10 (3.5 +/- 0.4 x 10(-3) s(-1), n = 6). It can be concluded that membrane potential changes in mature oligodendrocytes are accompanied by rapid changes in the K+ gradient resulting from K+ fluxes across the glial membrane. As a result of the reduced extracellular volume and increased tortuosity, the membrane fluxes produce larger changes in [K+]e in the more mature myelinated corpus callosum than before myelination. These conclusions also account for differences between membrane currents in cells in slices compared to those in tissue culture where the ECS is essentially infinite. The size and geometry of the ECS influence the membrane current patterns of glial cells and may have consequences for the role of glial cells in spatial buffering. PMID- 9211995 TI - Multiple class I motifs revealed by sequencing naturally processed peptides eluted from rat T cell MHC molecules. AB - Class I major histocompatibility complex (MHC) molecules interact with a diverse array of self and foreign peptides. Displayed on the cell surface, the class I/peptide complex provides an extracellular indication of the intracellular milieu. We have characterized the Lewis rat Vbeta8.2+ T cell hybridoma C14/BW12 12A1 by FACS analysis and have used immunoaffinity chromatography to purify class I molecules from these cells. Peptides eluted from the class I molecules have been fractionated by HPLC and sequenced. Self-peptide mixtures indicate two distinct peptide motifs, suggesting the possibility of multiple class I loci. The majority of the naturally processed peptide ligands were nonamers. Naturally processed peptide ligands fitting the first motif contained a hydrophobic leucine anchor residue at position three and a carboxyl-terminal serine anchor residue. A second motif was characterized by a tyrosine or phenylalanine residue at position three and a phenylalanine or isoleucine carboxyl-terminal residue. Four peptides derived from the Vbeta8.2 T cell receptor have sequences that fit these motifs, providing a mechanistic explanation for their immunoregulatory role. Identification of these class I peptide binding motifs will be useful for predicting potential CTL epitopes in studies on autoimmunity, immunoregulation and transplantation in the Lewis rat. PMID- 9211996 TI - Cure and follicular lymphoma. PMID- 9211997 TI - Is comprehensive lymphatic irradiation for low-grade non-Hodgkin's lymphoma curative therapy? Long-term experience at a single institution. AB - PURPOSE: This study reports 21 patients with Stage I-III low-grade non-Hodgkin's lymphoma who were treated with comprehensive lymphatic irradiation (CLI) at the University of Florida between 1966 and 1992. METHODS AND MATERIALS: Sites clinically involved with disease were treated with 30 Gy, whereas clinically uninvolved sites were treated with 25 Gy. Median follow-up for the group was 14 years (24.5 years for Stage III patients). RESULTS: Overall absolute survival rates at 5, 10, and 15 years were 84%, 68%, and 34%. Cause-specific survival rates at 5, 10, and 15 years were 84%, 68%, and 56%. Freedom-from-relapse rates at 5, 10, and 15 years were 75%, 58%, and 58%, with no relapses noted after 10 years. Bulky disease (>6 cm) was a significant indicator of poor prognosis for cause-specific survival (p = .01). CONCLUSION: These data support findings from other institutions suggesting a role for CLI as potentially curative therapy with acceptable toxicity and a short treatment time for patients with Stages I and II and limited Stage III disease. PMID- 9211998 TI - Quality of life and neuropsychological evaluation for patients with malignant astrocytomas: RTOG 91-14. Radiation Therapy Oncology Group. AB - With increasingly aggressive neurosurgical and radiation therapy modalities (gamma knife, external beam stereotactic radiation and interstitial brachytherapy with or without hyperthermia) offered to patients with malignant astrocytomas (MA), increasing national demand for medical outcome studies and rising health care costs amidst public, business, and governmental debate to cut spending, we as physicians are obligated to continue our research to find effective treatments for malignant astrocytoma (MA) and a cost-effective means to study their impact upon the patient's quality of life (QOL). PURPOSE: We report data that was collected within the Radiation Therapy Oncology Group (RTOG) on 126 patients with MA who were enrolled in RTOG 91-14. This study was undertaken to prospectively test the feasibility of performing quality of life (QOL) and neuropsychological evaluation (NPE) and collecting this data within the RTOG. RESULTS: The NPE and QOL parameters that were used in this study are cost effective. They are not only much cheaper than formal cognitive and memory testing, but also provide additional information regarding the patients' day to day functional abilities that are not provided by the current routinely used means, such as KPS. The Mini Mental Status Exam (MMSE) provides greater sensitivity to patients' differences in neurological status and may be preferable to NFS as an eligibility criteria. PMID- 9211999 TI - Neuropsychological sequelae of medulloblastoma in adults. AB - PURPOSE: To investigate the neuropsychological consequences of medulloblastoma in adults. METHODS: Patients 18 years of age or older who had medulloblastoma and at least 3 years of disease-free survival were eligible. A battery of tests was conducted to assess global intellectual functioning, verbal ability, visuospatial ability, memory, reasoning, and academic proficiency. For the verbal memory performance, each patient was matched with two normal controls selected on the basis of age, sex, and level of education. RESULTS: Review of the Neuro-Oncology database revealed 24 patients eligible for the study. Of these, 10 patients (6 good-risk and 4 poor-risk) agreed to participate; 7 patients were lost to follow up; 5 lived too far away to come to the testing site, and 2 refused testing. There were four men and six women; their mean age was 36.5 years at testing and 29.9 years at surgical diagnosis. Mean dose of whole brain radiation was 34.5 Gy. Mean interval between diagnosis and testing was 79.1 months. Test results demonstrated below average intelligence quotients (mean intelligence quotient 90.2; range 67-103) and specific deficits in memory, reasoning, visuospatial ability, and arithmetic. CONCLUSION: Adults with medulloblastoma in a prolonged disease-free status may suffer significant cognitive deficits. We recommend further controlled, prospective studies to evaluate cognitive outcomes in this patient population in the hope that interventional strategies could be developed, or treatment modified to minimize such toxicities. PMID- 9212000 TI - Corpus callosum involvement as a prognostic factor for patients with high-grade astrocytoma. AB - PURPOSE: To evaluate corpus callosum involvement as a prognostic factor for patients with high-grade astrocytoma. METHODS: From 1986 through 1994, 141 adult patients with Karnofsky performance status (KPS) > or = 40 underwent primary treatment for anaplastic astrocytoma (AA) or glioblastoma multiforme (GBM) at the University of Washington Medical Center. Preoperative magnetic resonance imaging and/or computed tomography to assess corpus callosum involvement was available for 105 of these patients. Corpus callosum involvement was evaluated as a prognostic factor for survival using recursive partitioning analysis and multivariate analysis with a Cox proportional hazards model. RESULTS: For the 105 patients evaluable for corpus callosum involvement, the median and 2-year survival were 59 weeks and 28%, respectively. On multivariate analysis, the only independent prognostic factors were KPS (p = 0.0001) and histology (p = 0.042). On recursive partitioning analysis, the first significant split occurred at KPS < 70 vs. KPS > or = 70. Patients with KPS > or = 70 were split by age (< 50 years vs. > or = 50 years), with those younger than 50 years further split by absence or presence of corpus callosum involvement. Among patients with KPS > or = 70 and age < 50 years, median survival was 57 weeks if the corpus callosum was involved (35% 2-year survival) and 105 weeks if the corpus callosum was not involved (56% 2-year survival). CONCLUSION: Corpus callosum involvement based on preoperative imaging is an unfavorable prognostic factor for survival among the subgroup of young, good-performance-status patients with high-grade astrocytoma. PMID- 9212001 TI - Short course radiotherapy is an appropriate option for most malignant glioma patients. AB - PURPOSE: To determine whether a shortened course of radiotherapy (RT) is an appropriate treatment option for malignant glioma patients. METHODS AND MATERIALS: Prognostic groups published by the Radiation Therapy Oncology Group (RTOG) are used to compare results for a short radiotherapy regimen with results of aggressive protocol treatment. The study group includes 219 patients treated during 1975-1993 with 51 Gy in 17 fractions. Patients were retrospectively assigned to six prognostic groups previously identified in a recursive partitioning analysis of the RTOG. The prognostic groups are based on age, histology, performance status, mental status, neurologic function, resection extent, length of symptoms, and RT dose. RESULTS: The six RTOG prognostic groupings were significantly predictive of outcome for patients treated with this shortened regimen (log-rank, p < 0.001). The median survival for our patients by RTOG groups 1-6 were 68, 57, 22, 13, 8, and 5 months, respectively. Two-year survival results were 64, 67, 45, 8, 3, and 3%. The median and two-year survival results for each prognostic grouping were similar to the results achieved by aggressive treatment on RTOG malignant glioma trials for selected patients. Treatment toxicity was uncommon. CONCLUSION: This shortened regimen is an appropriate treatment option for most malignant glioma patients (RTOG groups 4 6), resulting in similar survival as standard regimens with reduced patient effort and cost. Although acute side effects are acceptable and the risk of brain necrosis is low, we do not recommend this treatment to the minority of patients who have a substantial long term survival probability (RTOG groups 1-3) because long term neurocognitive assessment is lacking. PMID- 9212002 TI - Definitive radiotherapy for early glottic carcinoma: prognostic factors and implications for treatment. AB - PURPOSE: Treatment and disease-related factors were analyzed for their influence on the outcome of patients treated definitively with irradiation (RT) for early glottic carcinoma. METHODS AND MATERIALS: One hundred two patients with stage T1 or T2 glottic carcinomas were treated definitively with RT from December 1983 through September 1993. Median follow-up time was 63 months. Factors analyzed for each patient included age, sex, stage, anterior commissure involvement, surgical alternative, histologic differentiation, field size, total dose, fraction size, and total treatment time. Survival analysis methods were employed to assess the effects of these factors on local control and complication rates. RESULTS: The 5 year local control rates by stage were as follows: T1a, 92%; T1b, 80%; T2a, 94%; and T2b, 23%. By univariate analysis, factors found to have a significant impact on local control were stage, surgical alternative, fraction size, anterior commissure involvement, and overall treatment time. By multivariate analysis, stage, field size, and fraction size were the only significant factors that independently influenced local control. CONCLUSIONS: The inferior control rate for stage T2b lesions has implications for treatment. Our study supports the conclusions of reports in the literature showing that low fraction size negatively influences outcome in patients with early glottic cancer. PMID- 9212003 TI - Reirradiation for recurrent nasopharyngeal carcinoma: factors affecting the therapeutic ratio and ways for improvement. AB - PURPOSE: To identify factors for maximizing local salvage and minimizing damages by reirradiation for recurrent nasopharyngeal carcinoma. METHODS AND MATERIALS: 654 patients with recurrent nasopharyngeal carcinoma treated by reirradiation during 1976-1992 were retrospectively analyzed. Various fractionation schedules had been used during primary treatment with the total dose ranging from 45.6-70 Gy, fractional dose (at different phases) 1.5-4.2 Gy, and overall time 36-101 days. The gap between the two courses ranged from 0.5-10.6 years. Eighty-two percent of patients were reirradiated with teletherapy, 6% brachytherapy, and 12% with both. For those treated with teletherapy alone, the total dose ranged from 7.5-70 Gy, fractional dose 1.8-5 Gy, and overall time 3-89 days. RESULTS: The 5 year actuarial local salvage and complication-free rates were 23% and 52%, respectively. Multivariate analyses showed that the extensiveness of local recurrence was the most significant factor affecting local salvage, while T-stage of primary tumor also influenced prognosis. Choice of method for reirradiation and fractional effect during both courses affected the risk of late complications. For patients treated by teletherapy alone, the hazard of local failure decreased by 1.7% per Biological Effective Dose (assuming alpha/beta ratio = 10) of the second course, while radiation factors during primary radiotherapy had no significant effect. On the other hand, the risk of late complications was predominantly affected by the primary treatment: the hazard increased by 4.2% per Biological Effective Dose (assuming alpha/beta ratio = 3) of the first course, while the corresponding impact of reirradiation failed to reach statistical significance. Length of the gap between the two courses did not affect the outcome. CONCLUSION: Early detection of local recurrence and adequate total dose by reirradiation are crucial for improving the chance of local salvage. Combination of teletherapy and brachytherapy should be considered whenever feasible and large fractional dose avoided to minimize late complications. Optimization of biological dose during primary treatment is important. PMID- 9212004 TI - Salvage brachytherapy of posterior pharyngeal wall squamous cell carcinoma in a previously irradiated area. AB - PURPOSE: Brachytherapy performed in patients with posterior pharyngeal wall carcinoma in a previously irradiated area is evaluated in terms of local control, survival, and complications. METHODS AND MATERIALS: Between January 1982 and July 1993, 14 patients were treated with interstitial low dose rate brachytherapy alone for posterior pharyngeal wall squamous cell carcinoma in a previously irradiated area (local recurrences in five cases and second tumors in nine cases). Tumor size ranged from 1 to 4 cm. No patient had a macroscopic nodal involvement or metastase at the time of diagnosis. Median dose delivered was 55 Gy (39 to 60 Gy). RESULTS: Thirteen patients were assessed for local control. Twelve of them achieved complete macroscopic response within 2 months after brachytherapy. Local relapse occurred in five patients, from 5 to 29 months after brachytherapy. One patient developed distant metastatis without loco-regional relapse. Disease free survival was 69, 59, and 37% at 1, 2, and 5 years, respectively; overall survival was 78, 50, and 21% at 1, 2, and 5 years, respectively. Three patients were still alive without recurrence (8, 8, and 10 years after treatment). We did not observe any severe acute or delayed toxicity. CONCLUSION: Based on these results, interstitial brachytherapy should be considered as a potentially curative treatment for selected patients with posterior pharyngeal wall squamous cell carcinoma in a previously irradiated area. There are no reports in the literature on this subject. PMID- 9212005 TI - Chronic rectal bleeding after high-dose conformal treatment of prostate cancer warrants modification of existing morbidity scales. AB - PURPOSE: Serious late morbidity (Grade 3/4) from the conformal treatment of prostate cancer has been reported in <1% to 6% of patients based on existing late gastrointestinal (GI) morbidity scales. None of the existing morbidity scales include our most frequently observed late GI complication, which is chronic rectal bleeding requiring multiple fulgerations. This communication documents the frequency of rectal bleeding requiring multiple fulgerations and illustrates the variation in reported late serious GI complication rates by the selection of morbidity scale. METHODS AND MATERIALS: Between May 1989 and December 1993, 352 patients with T1-T3 nonmetastatic prostate cancers were treated with our four field conformal technique without special rectal blocking. This technique includes a 1-cm margin from the clinical target volume (CTV) to the planning target volume (PTV) in all directions. The median follow-up for these patients was 36 months (range 2-76), and the median center of prostate dose was 74 Gy (range 63-81). Three morbidity scales are assessed: the Radiation Therapy Oncology Group (RTOG), the Late Effects Normal Tissue Task Force (LENT), and our modification of the LENT (FC-LENT). This modification registers chronic rectal bleeding requiring at least one blood transfusion and/or more than two coagulations as a Grade 3 event. Estimates for Grade 3/4 late GI complication rates were determined using Kaplan-Meier methodology. The duration of severe symptoms with chronic rectal bleeding is measured from the first to the last transrectal coagulation. Latency is measured from the end of radiotherapy to surgery, first blood transfusion, or third coagulation procedure. RESULTS: Sixteen patients developed Grade 3/4 complications by one of the three morbidity scales. Two patients required surgery (colostomy or sigmoid resection), three required multiple blood transfusions, two required one or two blood transfusions, and nine required at least three coagulations. The median duration of bleeding for those patients requiring multiple procedures was 7 months (range 3-33) and the median latency was 22 months (range 9-40). The 5-year actuarial rate of Grade 3/4 complications by each scale are: RTOG 0.7%, LENT 2%, and FC-LENT 6%. The rate of chronic rectal bleeding increases with increasing dose and is low in patients treated with conventional techniques owing to lower doses. CONCLUSION: Chronic rectal bleeding requiring any blood transfusion(s) or multiple coagulation procedures is our most frequently observed complication. This complication appears late in follow-up and is present for a long duration. We believe this justifies the inclusion of chronic rectal bleeding requiring multiple coagulation procedures as a Grade 3 event in future morbidity scales. Our data illustrate that published Grade 3/4 morbidity rates are highly dependent on the morbidity scale selected, as our data show 0.7% RTOG, 2% LENT, and 6% FC-LENT. Obviously, a uniform scale is required that includes the newly recognized serious late effects associated with the conformal treatment of prostate cancer. PMID- 9212006 TI - Low acute gastrointestinal and genitourinary toxicities in whole pelvic irradiation of prostate cancer. AB - PURPOSE: This retrospective study was done to determine the frequency and severity of acute gastrointestinal (GI) and genitourinary (GU) toxicity associated with whole pelvic radiotherapy of localized prostate cancer. METHODS AND MATERIALS: Between 1989 and 1994, we treated 156 patients with localized prostate cancer, ranging in age from 54 to 86 (median 71), of which 86 were older than 70 years of age. No attempt at selection was made, and many were from the Veteran's Administration Hospital where they had been precluded from their surgical program because of comorbidities and/or advanced age. Of 156 patients, 45 (28.8%) underwent pretreatment laparoscopic lymphadenectomy (LAP); 40 had negative findings. Four-field box technique was used for all patients. We treated the whole pelvis to 45 Gy, followed by a cone-down and a final boost to a total dose of 72 Gy. The cone-down to the lower pelvis and boost to the prostate were based on computed tomography and/or magnetic resonance imaging findings for volume reconstruction with field size of approximately 8 x 8 and 6 x 6 cm, respectively. Diet instructions were given before treatment and emphasized weekly. Toxicities were evaluated weekly by physicians and nurses independently using Cancer and Leukemia Group B (CALGB) grading criteria. RESULTS: The acute GI and GU toxicities gradually increased from Week 2, peaked at Week 5, and then declined after that. During Week 5, acute Grade 1-3 GI morbidities were observed in 19 (12.2%), 2 (1.3%), and 1 (0.6%) patients, respectively. Genitourinary toxicity was similar, accounting for 17 (10.9%), 6 (3.8%), and 1 (0.6%), respectively. Overall Grade 2 toxicities occurred in 30 of 156 patients (19%). Comorbidity was associated with more GI toxicity. Patients over 70 years of age tended to reach the maximal GI and GU toxicity 1-2 weeks earlier than did patients under the age of 70. Of the patients who did not follow the diet instruction, all experienced side effects. CONCLUSIONS: Whole pelvic irradiation was very well tolerated in this group of patients. The frequency of acute Grade 2 GI and GU toxicity compared favorably with the reported results of conformal treatment. Diet restriction and psychosocial input may have had a positive impact. Late sequelae will be evaluated in long-term follow-up. PMID- 9212007 TI - Variation in prostate position quantitation and implications for three dimensional conformal treatment planning. AB - PURPOSE: This study describes and quantitates the motion, i.e., variation in position, of the prostate within the pelvis and its effect on target and normal organ dose. METHODS AND MATERIALS: The motion of the planning target volume (PTV) borders and center of mass was studied in 13 patients with carcinoma of the prostate through the use of superimposed serial computerized tomography (CT) scans. Changes in bladder and rectal volumes were measured and their relationship to displacements of the PTV position were noted. The effects of this motion on target and normal organ doses were measured. RESULTS: A variability in the position of the PTV is seen over time, which is related to changes in bladder and rectal volumes. The one standard deviation displacements of the PTV center of mass with respect to the planning scan center of mass position were 0.12, 0.40, and 0.31 cm in the lateral, anterior-posterior, and superior-inferior directions, respectively. Movement was significantly larger in the superior part of the PTV above the base of the bladder than in the inferior part. Movement of the borders of the PTV outward from the patient axis; hence, toward the edges of the treatment field, was also examined. Outward displacements of the anterior target border below the base of the bladder were less than 0.3 cm in 90% of the cases, and 1.4 cm above the bladder base. For the posterior wall these displacements were less than 0.7 cm and 1.1 cm, respectively, whereas the lateral border displacements were less than 0.3 cm throughout (90% confidence limits). These displacements would cause a median of 6% of the PTV to receive less than 95% of the planned dose for any given treatment day in these patients; the effect on rectal and bladder wall doses was greater and true doses may not be measurable through the use of only one treatment planning CT scan. CONCLUSIONS: The prostate is not a static organ, but rather has some limited motion in the pelvis secondary to bladder and rectal volume changes. This motion has been quantified for a group of patients, and may provide a guide to further studies on the placement of field borders. PMID- 9212009 TI - External beam abdominal radiotherapy in patients with seminoma stage I: field type, testicular dose, and spermatogenesis. AB - PURPOSE: To establish a predictive model for the estimation of the gonadal dose during adjuvant para-aortic (PA) or dog leg (DL: PA plus ipsilateral iliac) field radiotherapy in patients with testicular seminoma. METHODS AND MATERIALS: The surface gonadal dose was measured in patients with seminoma Stage I receiving PA or DL radiotherapy. Sperm cell analysis was performed before and 1 year after irradiation. PA and DL radiotherapy were simulated in the Alderson phantom while we measured the dose to the surface and middle of an artificial testicle, varying its position within realistic anatomical constraints. The symphysis-to-testicle distance (STD), field length, and thickness of the patient were experimental variables. The developed mathematical model was validated in subsequent patients. RESULTS: The mean gonadal dose in patients was 0.09 and 0.32 Gy after PA and DL irradiation, respectively (p < 0.001). DL radiotherapy, but not PA irradiation led to significant reduction of the sperm count 1 year after irradiation. The gonadal dose-reducing effect of PA irradiation was confirmed in the Alderson phantom. A significant correlation was found between the STD and the gonadal dose during DL irradiation. A mathematical model was established for calculation of the gonadal dose and confirmed by measurements in patients. CONCLUSIONS: During radiotherapy of seminoma, the gonadal dose decreases with increasing STD. It is possible to predict the individual gonadal dose based on delivered midplane dose and STD. PMID- 9212008 TI - A study of reproductive function in patients with seminoma treated with radiotherapy and orchidectomy: (SWOG-8711). Southwest Oncology Group. AB - PURPOSE: The results of Southwest Oncology Group Study 8711 (Group 2B) are presented. The objective was to evaluate the natural history of sperm concentration and selected hormonal parameters in patients with testicular cancer treated with orchiectomy and radiotherapy. METHODS AND MATERIALS: Of a total of 207 patients enrolled on SWOG 8711, 53 pure seminoma patients were identified who were treated with orchiectomy and radiotherapy only. Sperm concentration, follicle-stimulating hormone (FSH) levels, and sexual satisfaction scores were the main parameters followed. RESULTS: A fraction of the patients were infertile prior to receiving radiotherapy. Our analysis indicates that incidental radiation dose to the remaining testicle affects time to recovery of fertility, and at an aggregate level, changes in FSH mirror changes in sperm concentration over time. This phenomenon is the same as that described in patients free from testicular cancer. These men evaluated their sexual activity as good after orchidectomy. CONCLUSION: Our data support the use of clamshell-type testicular shields as a means of providing maximum protection to the remaining testicle. PMID- 9212011 TI - Impact of setup variability on incidental lung irradiation during tangential breast treatment. AB - PURPOSE: This study aimed to determine the variability in treatment setup during a 5-week course of tangential breast treatment for patients immobilized in a customized hemibody cradle, to assess the relationship between the height of the lung shadow on the tangential port film and the percentage of lung volume irradiated, and to estimate the impact of setup variabilities on irradiated lung volume. METHODS: One hundred seventy-two port films were reviewed from 20 patients who received tangential beam treatment for breast cancer. The height of the lung shadow at the central axis (CLD) on each port film was compared to the corresponding simulator film as an assessment of setup variability. A three dimensional dose calculation was performed, and the percentage of total lung volume within the field was correlated with the CLD. The three-dimensional dose calculation was repeated for selected patients with the location of the treatment beams modified to reflect typical setup variations. RESULTS: The CLD measured on the port films was within 3 mm of that prescribed on the simulator film in 43% (74 of 172) of the port films. The variation was 3-5 mm in 26%, 5-10 mm in 25%, and >10 mm in 6%. The height of the lung shadow correlated with the percentage of lung volume included in the radiation field (r2 = 0.6). Typical variations in treatment setup resulted in < or = 5% fluctuation in the absolute volume of ipsilateral lung irradiated. CONCLUSION: The current immobilization system used in our clinic provides a clinically acceptable reproducibility of patient setup. The height of the lung shadow is reasonably well correlated with the percentage of irradiated lung volume. During a typical 5-week course of radiotherapy, the ipsilateral irradiated lung volume fluctuates <5%. PMID- 9212010 TI - Trends in primary surgical and radiation therapy for localized breast cancer in the Detroit Metropolitan area 1973-1992. AB - PURPOSE: The purpose of this report is to describe trends in primary surgical and radiation therapy for localized breast cancer from 1973 through 1992 among residents of the Detroit Metropolitan area. METHODS AND MATERIALS: Data on surgical and radiation therapy procedures for women with local stage breast cancer were obtained from the population-based Metropolitan Detroit Cancer Surveillance System (MDCSS). RESULTS: Women age 75 years and older were treated less aggressively than younger women (< age 75) as evidenced by higher rates of simple mastectomy or no treatment among older women. Younger women (< age 75) were more likely to have had optimal breast conservation therapy which consisted of partial mastectomy, axillary lymph node dissection (ALND), and radiation therapy, than were women who were older than 75. Partial mastectomy has increased proportionally from 4% of all breast cancer surgeries in the time period 1973 to 1977, to 39% of all surgeries from 1988 through 1992. CONCLUSION: A marked difference in surgical treatment of breast cancer exists for younger vs. older women. Despite changes in surgical treatment trends for breast cancer, a large proportion of women who are candidates for conservative therapy continue to undergo mastectomy. PMID- 9212012 TI - Influence of modifications in breast irradiation technique on dose outside the treatment volume. AB - PURPOSE: There is increasing interest in potential long-term effects of radiotherapy (RT) in patients treated for breast cancer, particularly those in whom long-term survival can be expected. The purpose of the present study was to determine the effects of treatment techniques, including patient positioning (supine vs. prone) on the absorbed dose in organs at a distance from the treatment volume in breast RT. METHODS AND MATERIALS: Dose distribution was studied in a Rando-Alderson phantom, modified with a simulated left breast of tissue-equivalent material. RT delivery was studied using 60Co and 6 MV x-ray beams, as well as electrons and a 192Ir source for tumor bed boost RT. Doses were measured in several organs and tissues of interest using LiF thermoluminescent dosimeters. Tangential breast RT was simulated using both supine and prone positioning. RESULTS: Peripheral doses generally decreased approximately exponentially with distance from the edge of the treatment field. Peripheral doses in various target organs were significantly higher for supine than for prone tangential breast RT (for 50 Gy prescribed dose): 0.50 Gy vs. 0.25 Gy for the upper abdomen, 0.05 Gy vs. 0.02 Gy for pelvic organs, 0.17 Gy vs. 0.08 Gy for active bone marrow, and 0.47 Gy vs. 0.12 Gy for ipsilateral lung (discounting lung in primary beam). CONCLUSIONS: The present study suggests that peripheral doses in several organs and tissues of interest can be reduced by 40 to 75% by prone tangential breast RT. These results may have implications for future strategies in the treatment of screen-detected breast cancer. PMID- 9212013 TI - American Brachytherapy Society (ABS) consensus guidelines for brachytherapy of esophageal cancer. Clinical Research Committee, American Brachytherapy Society, Philadelphia, PA. AB - INTRODUCTION: There is wide variation in the indications, treatment regimens, and dosimetry for brachytherapy in the treatment of cancer of the esophagus. No guidelines for optimal therapy currently exist. METHODS AND MATERIALS: Utilizing published reports and clinical experience, representatives of the Clinical Research Committee of the American Brachytherapy Society (ABS) formulated guidelines for brachytherapy in esophageal cancer. RESULTS: Recommendations were made for brachytherapy in the definitive and palliative treatment of esophageal cancer. (A) Definitive treatment: Good candidates for brachytherapy include patients with unifocal thoracic adeno- or squamous cancers < or = 10 cm in length, with no evidence of intra-abdominal or metastatic disease. Contraindications include tracheal or bronchial involvement, cervical esophagus location, or stenosis that cannot be bypassed. The esophageal brachytherapy applicator should have an external diameter of 6-10 mm. If 5FU-based chemotherapy and 45-50-Gy external beam are used, recommended brachytherapy is either: (i) HDR 10 Gy in two weekly fractions of 5 Gy each; or (ii) LDR 20 Gy in a single course at 0.4-1 Gy/hr. All doses are specified 1 cm from the midsource or mid-dwell position. Brachytherapy should follow external beam radiation therapy and should not be given concurrently with chemotherapy. (B) Palliative treatment: Patients with adeno- or squamous cancers of the thoracic esophagus with distant metastases or unresectable local disease progression/recurrence after definitive radiation treatment should be considered for brachytherapy with palliative intent. After limited dose (30 Gy) EBRT, the recommended brachytherapy is either: (i) HDR 10-14 Gy in one or two fractions; or (ii) LDR 20-25 Gy in a single course at 0.4-1 Gy/hr. The need for external beam radiation in newly diagnosed patients with a life expectancy of less than 3 months is controversial. In these cases, HDR of 15 20 Gy in two to four fractions or LDR of 25-40 Gy at 0.4-1 Gy/hr may be of benefit. CONCLUSION: ABS guidelines for esophageal brachytherapy now exist and will be updated by the ABS in the future, as clinical data using more uniform treatment techniques becomes available. PMID- 9212015 TI - Hyperfractionated external radiation therapy in stage IIIB carcinoma of uterine cervix: a prospective pilot study. AB - PURPOSE: Brazil has one of the highest incidence of carcinoma of the cervix in the world. Half of the patients have advanced stages at the diagnosis. Due to this large number of patients we decided to conduct a prospective pilot study to investigate the tolerance to and survival rate with hyperfractionated external radiotherapy only in patients with Stage IIIB carcinoma of the uterine cervix. METHODS AND MATERIALS: Between January 1991 and December 1993, 23 patients underwent hyperfractionated external beam radiotherapy without brachytherapy. All cases were biopsy proven squamous cell carcinoma of cervix clinically Staged as IIIB (FIGO). Hyperfractionation (HFX) was given with 1.2 Gy doses, twice daily at 6-h interval, 5 days/week, to the whole pelvis up to 72 Gy within 30 working days. Complications were evaluated by an adaptation ot the RTOG Radiation Morbidity Scoring Table graded as 1 = none/mild; 2 = moderate, and 3 = severe. RESULTS: Follow-up ranged from 27 to 50 months (median 40 months) on the 9 to 23 living patients at the time of the analysis in December 1995. There was no severe acute toxicity, but moderate acute reaction was high: 74%. The commonest site of complication was the intestine where severe late toxicity occurred in 2 of 23 (9%). Overall survival rate at 27 months was 48% and at 40 months was 43%. DISCUSSION: There is little information in literature about HFX in carcinoma of the cervix. This is the third published study about it and the one that gave the highest total dose with external HFX of 60 x 1.2 Gy = 72 Gy. Theoretically, through the linear quadratic formula this schedule of HFX would be equivalent to 30 x 2 Gy = 60 Gy of standard fractionation, both treatments given in 30 working days. HFX schedules must be tested to establish their safety. Present results suggest being possible to further increase the total dose in the pelvis with hyperfractionated irradiation. PMID- 9212014 TI - Continuous hyperfractionated accelerated radiotherapy (CHART) in localized cancer of the esophagus. AB - PURPOSE: To assess the efficacy and toxicity of continuous hyperfractionated accelerated radiotherapy (CHART) in locoregional control compared with a historical group of patients treated with conventionally fractionated radical radiotherapy. METHODS AND MATERIALS: Between 1985 and 1994, 54 patients with localized esophageal cancer were treated with CHART. Twenty-eight patients received CHART alone (54 Gy in 36 fractions over 12 consecutive days) and 15 were given intravenous mitomycin C and cisplatin on days 10 and 13, respectively. Eleven patients received 40.5 Gy in 27 fractions over 9 days, followed by a single high-dose-rate intraluminal brachytherapy insertion of 15 Gy at 1 cm. RESULTS: Acute toxicity was well tolerated and dysphagia was improved in 35 patients (65%), with 28 (52%) eating a normal diet by week 12. This compares with an improvement in dysphagia score in 72% of the conventionally treated group. The median duration of relief of dysphagia was 7.8 months (range 0-41.4) in the CHART group compared with 5.5 months (range 0-48) in the controls. Strictures developed in 29 patients (61%) and 18 were confirmed on biopsy to be due to recurrent disease. Median survival was 12 months (range 0.5-112) in the CHART group and 15 months (range 3.6-56) in the control patients. CONCLUSION: CHART is well tolerated and achieves a high rate of local control. Palliation in the short overall treatment time of esophageal cancer is an advantage in these patients whose median survival is only 12 months. PMID- 9212016 TI - Pelvic exenteration for cervix cancer: would additional intraoperative interstitial brachytherapy improve survival? AB - OBJECTIVE: Improved local control with the addition of brachytherapy to pelvic exenteration for recurrent cervical cancer has been reported to improve survival. We examined the sites of recurrence after pelvic exenteration to determine if these patients might have been salvaged by the improved local control promised by interstitial brachytherapy. We sought to identify risk factors available intraoperatively or perioperatively which might predict decreased local control. METHODS: A retrospective review of 26 patients with recurrent cervical cancer who underwent total pelvic exenteration since 1988 at our institution was performed. RESULTS: Overall, the mean follow-up was 29.5 months (range 6.1-81.6). Of the 26 patients, 14 had no evidence of disease (NED), 1 was alive with disease (AWD), 9 were dead of disease (DOD), and 2 died of unrelated causes (DOC). Seven of 26 patients (27%) had margins < or = 5 mm, of whom 2 were NED, 4 DOD, and 1 AWD. Seven of 26 (27%) patients had lymphovascular involvement (LVI) or perineural invasion (PNI) with clear margins. Three of the seven with LVI or PNI and clear margins were NED, and four DOD. Of the 10 failures, 9 (90%) had close margins, PNI, or LVI. CONCLUSION: Our data reveal that 9 of 14 (64%) patients with close margins, LVI, or PNI were DOD or AWD, and 6 of 9 of those patients suffered local regional failure alone. Brachytherapy has the potential to cure 6 of 14 (43%) patients with these risk factors. Further study of brachytherapy at the time of pelvic extenteration is warranted. PMID- 9212017 TI - Randomized study of chemotherapy/radiation therapy combinations for favorable patients with locally advanced inoperable nonsmall cell lung cancer: Radiation Therapy Oncology Group (RTOG) 92-04. AB - PURPOSE: The purpose of this study was to compare the severity and distribution of the toxicities associated with the two different combinations of chemotherapy and radiotherapy. METHODS AND MATERIALS: This prospective randomized trial studied toxicities associated with induction chemotherapy followed by concurrent treatment (Arm 1) vs. immediate concurrent chemotherapy/radiotherapy (CT/RT) (Arm 2). Arm 1 consisted of vinblastine (VB), 5 mg/M2 I.V. bolus weekly, weeks 1-5 and cisplatin (DDP), 100 mg/M2 days 1 and 29, DDP 75 mg/M2, days 50, 71, and 92. Daily RT started on day 50; a total dose of 63 Gy was given in 34 fractions in 7 weeks. In Arm 2 RT started day 1; a total dose of 69.6 Gy was given in 58 fractions of 1.2 Gy bid, 5 days per week for 6 weeks with DDP 50 mg/M2 i.v. days 1 and 8, and oral VP-16 50 mg b.i.d. during the first 10 days of RT. DDP/VP-16 were repeated beginning day 29. Survival was used as the Phase II endpoint. RESULTS: Between July 1992 and February 1994, 168 patients were randomized; 162 evaluable patients had minimum follow-up of 20 months. Eighty patients were registered to Arm 1 and 82 to Arm 2. Pretreatment characteristics were distributed evenly. Arm 1 had significantly more Grade 4 hematologic toxicity (62%) than Arm 2 (33%) (p = 0.021). Acute nonhematologic Grade 3+ toxicity was also greater (p = 0.018) in Arm 2 than Arm 1 due mainly to esophagitis (38 vs. 6%; p < 0.0001). Grade 3+ late esophageal toxicity was 12% on Arm 2 compared to 3% on Arm 1 (p = 0.006). There were no differences between the two arms in compliance with protocol specifications for either RT or CT. At 1 year, 31.7% of patients had in-field progression on Arm 1 compared to 19.8% on Arm 2 (p = 0.042), but overall progression-free survival rates were nearly identical; 50 and 49% for Arms I and II, respectively, at 12 months. One-year and median survivals were 65% and 15.5 months on Arm 1 compared to 58% and 14.4 months on Arm 2. CONCLUSION: Whereas hematologic toxicity was greater in Arm 1, esophageal toxicity, both acute and late, was greater in Arm 2. Infield progression was lower in Arm 2, but overall progression rates were similar and there were no significant differences in survival between the two arms. A 3-arm randomized Phase III study is underway in the RTOG to compare sequential and concurrent CT/RT. PMID- 9212018 TI - Concurrent carboplatin, etoposide and thoracic radiation for poor-risk stage III non-small-cell lung carcinoma: a pilot study. AB - PURPOSE: A pilot study was conducted to assess the tolerance and efficacy of concurrent carboplatin, etoposide, and thoracic radiation in poor-risk patients with Stage III non-small-cell lung carcinoma (NSCLC). METHODS AND MATERIALS: Patients had Stages IIIA/IIIB NSCLC and were ineligible for available clinical trials employing cisplatin-based chemoradiation due to one or more protocol defined poor-risk factors or concomitant medical conditions. Treatment consisted of thoracic radiation, 1.8 to 2 Gy daily, to the primary tumor and regional lymph nodes to a total dose of 61 Gy. Concurrently, patients received carboplatin 200 mg/m2/day intravenously on days 1, 3, 29, and 31, and etoposide 50 mg/m2/day intravenously on days 1-4 and 29-32. Response was assessed by chest computed tomography (CT) 4 weeks after treatment was completed. RESULTS: A total of 26 patients were enrolled and 23 of these patients, including 11 with Stage IIIA and 12 Stage IIIB NSCLC, were eligible and assessable. Ninety-six percent (96%) of the patients completed the two planned courses of chemotherapy, and 87% completed the planned chest radiation. Grade III/IV toxicities included neutropenia in nine patients (39%), thrombocytopenia in five (22%), esophagitis in seven (30%), and nausea in two (9%). One patient died of a pulmonary embolism during treatment, and another died of complications due to a tracheoesophageal fistula. Four patients (17%) achieved a complete response and 16 (70%) a partial response, yielding an overall response rate of 87%. The median survival was 12 months, and the 2-year actuarial survival was 40%. CONCLUSION: This treatment regimen was well tolerated, with promising response and survival in poor-risk patients with Stage III NSCLC. These results are being validated in a Phase II trial conducted by the Southwest Oncology Group. PMID- 9212020 TI - Difluoromethylornithine enhanced uptake of tritiated putrescine in 9L rat brain tumors. AB - Difluoromethylornithine (DFMO) depletes endogenous putrescine and enhances the uptake of and retention of [3H] putrescine in vitro. To determine if DFMO also enhances uptake of [3H] putrescine in vivo, DFMO and trace doses of [3H] putrescine, dissolved in artificial CSF, were infused into growing (6-9 day) 9L brain tumors by means of osmotic pumps. When 7-day osmotic pumps were loaded with 1 microCi [3H] putrescine, with or without 10 or 100 mM DFMO, pumped at 1 microl/h, the mean uptake after 3 days was 168 +/- 62 cpm/mg tumor (17 rats) without DFMO, 300 +/- 197 cpm/mg tumor (11 rats) with 10 mM DFMO and 1088 +/- 421 cpm/mg tumor (11 rats) with 100 mM DFMO (p < or = 0.05 vs. control). Significantly less radioactivity was detected in the contralateral brain and in nonbrain tissues (0.5 +/- 0.1 to 14 +/- 5 cpm/mg). To measure the extent of [3H] putrescine distribution in the tumor, the same dose of drugs was delivered for a longer period of time, using 14-day pumps to allow tumors to become large enough to be divided into 1.4 mm thick transections. The mean radioactivity in the sections from eight control rats receiving [3H] putrescine without DFMO were not significantly different between the sections (174 +/- 61 cpm/mg tumor for sections containing the cannulas, 273 +/- 61 and 259 +/- 91 cpm/mg for adjacent sections). In the six rats given 100 mM DFMO there was a significant increase in mean radioactivity in the cannula containing section (2251 +/- 919 cpm/mg tumor). Mean counts from adjacent sections in these rats were 97 +/- 44 and 33 +/- 13 cpm/mg. Values for contralateral corpus striatum and nonbrain tissues ranged from 0.7 +/- 0.3 to 4.3 +/- 1.5 cpm/mg tissue. When DFMO was delivered directly to the tumors while [3H] putrescine was infused intraperitoneally, the uptake in the tumor slices was low (5-10 cpm/mg in different slices). These results demonstrate that infusion of DFMO directly into growing 9L brain tumors can selectively enhance the uptake of exogenous [3H] putrescine by rapidly dividing cells which are within a 1.4 mm diameter area at the cannula tip. Although these studies used [3H] putrescine at trace doses, it is estimated that infusion of higher doses of [3H] putrescine plus DFMO will selectively kill tumor cells. PMID- 9212019 TI - A phase II trial of radiochemotherapy with daily carboplatin, after induction chemotherapy (carboplatin and etoposide), in locally advanced nonsmall-cell lung cancer: final analysis. AB - PURPOSE: To evaluate feasibility and efficacy of concomitant radiochemotherapy (CRCT) in Stage IIIB nonsmall-cell lung cancer (NSCLC), two induction chemotherapy cycles combining etoposide and carboplatin were first delivered, followed by CRCT with daily radiation fraction in association with carboplatin. METHODS AND MATERIALS: Forty patients with biopsy-proven, locally advanced unresectable nonmetastatic NSCLC were enrolled. Induction chemotherapy consisted of two cycles (day 1 and day 28) of etoposide (VP16:100 mg/m2, days 1 to 3) and carboplatin (CBDCA:350 mg/m2, day 1). Irradiation starting at day 56, delivered 66 Gy in 2 Gy daily fraction, 5 days a week, along with a daily dose of CBDCA (15 mg/m2) given intravenously 2 to 4 h before radiation. In nonprogressive patients under induction chemotherapy, two additional cycles of VP16-CBDCA were administered 4 weeks after the completion of CRCT. RESULTS: Out of the 40 patients enrolled (38 males, 2 females), 37 (93%) received induction chemotherapy as scheduled, with 38% Grade 3-4 hematological toxicity. Response rate to induction chemotherapy was 11% (4/37). No tumor became resectable. CRCT was delivered to 32 of these 37 patients, with full doses given to 91% of them. Clinical and hematological Grade 3-4 toxicity rates were 21 and 13%, respectively. Additional chemotherapy was delivered in 12 of 26 nonprogressive patients. At final evaluation, performed 3 months after the end of CRCT, 38% of 26 evaluable patients were responders (4 complete and 6 partial), leading to a 25% (10 of 40) overall objective response rate. Of these 10 responders, 8 became responders after CRCT only. Overall, the 1-year local control rate was 28% (11 of 40). The median survival time was 9 months and the 1-year and 2-year overall survival rates were 38 and 15%, respectively. Thirty-six patients died from local progression (25 patients), distant metastasis (9 patients), or pulmonary fibrosis (2 patients). CONCLUSION: Concomitant CRCT with CBDCA is feasible with acceptable induction chemotherapy-related toxicity and a 1-year local control rate of 28%. Response rate to induction chemotherapy was low and better chemotherapy combination should be used to reduce distant failure probability and to improve local response rate before CRCT. PMID- 9212021 TI - Cell proliferation and death in the irradiated pituitary gland and its modification by growth stimulants. AB - PURPOSE: This study was undertaken to show whether the rate of expression of radiation injury in the rat pituitary gland could be accelerated by the use of growth stimulants. METHODS AND MATERIALS: Rat pituitary glands were irradiated in situ with a range of single doses up to 20 Gy. The rats were then given subcutaneous slow-release implants containing 17beta-estradiol (E2) and sulpiride (S) to stimulate lactotroph proliferation. Two sequential cycles were used, each consisting of stimulation (3 weeks) and withdrawal (2 weeks). Measurements were made of gland weight; BrdU-labeled, giant, and apoptotic cells; lactotrophs; as well as pituitary prolactin content, in response to exogenous thyroid-releasing hormone (TRH). RESULTS: The two cycles of stimulation/withdrawal resulted in marked changes in gland weight, BrdU-labeling index, and serum prolactin (PRL) levels in unirradiated rats. The proportion of immunopositive growth-hormone producing (GH) cells increased after irradiation. Radiation inhibited the hypertrophic response to E2 + S and also inhibited increases in BrdU-labeling index and serum PRL levels. Also, giant lactotrophs were observed in the irradiated pituitaries. However, they were not seen in the unirradiated rats or in the irradiated rats treated with E2 + S. TRH promoted PRL secretion in the unirradiated rat. In contrast, TRH inhibited PRL secretion in the irradiated rat and in all treatment groups receiving E2 + S. Apoptosis was induced by irradiation and was substantially increased in lactotrophs and in other cell types by withdrawal of the E2 and S stimulus, although the highest observed incidence was only 7 per 10,000 cells. CONCLUSION: Both irradiation and E2 + S treatment removed the hypothalamic control of PRL secretion, which reveals this important inhibitory action of TRH upon PRL secretion. This suggests that it is not suitable as a dynamic test of pituitary PRL reserves in such abnormal situations, where there may also be damage to the hypothalamic-pituitary vasculature. The increasing proportion of GH cells after irradiation indicates that lactotrophs respond more rapidly to irradiation. The stimulation by E2 + S somehow prevented the radiation-damaged lactotrophs from becoming giant cells. Also, the ratio of apoptotic cells to BrdU-labeled cells was increased by the E2 + S treatment, indicating that the E2 + S did enhance radiation-induced cell death relative to cell renewal. However, overall, the E2 + S stimulus protocol did not promote a dramatic increase in cell death (apoptosis) nor a marked decrease in residual gland weight after irradiation. Hence, its use would probably not be beneficial in the treatment of slow-responding prolactinomas, if malignant lactotrophs respond similarly to the normal pituitary lactotrophs. However, the observation of induced apoptosis after hormone and drug withdrawal suggests that agents which promote tumor shrinkage may be effective by causing rapid apoptosis of tumor cells in vivo. PMID- 9212023 TI - Cellular radiosensitivity of small-cell lung cancer cell lines. AB - PURPOSE: The objective of this study was to determine the radiobiological characteristics of a panel of small-cell lung cancer (SCLC) cell lines by use of a clonogenic assay. In addition, we tested whether comparable results could be obtained by employing a growth extrapolation method based on the construction of continuous exponential growth curves. METHODS AND MATERIALS: Fifteen SCLC cell lines were studied, applying a slightly modified clonogenic assay and a growth extrapolation method. A dose-survival curve was obtained for each experiment and used for calculating several survival parameters. The multitarget single hit model was applied to calculate the cellular radiosensitivity (D0), the capacity for sublethal damage repair (Dq), and the extrapolation number (n). Values for alpha and beta were determined from best-fit curves according to the linear quadratic model and these values were applied to calculate the surviving fraction after 2-Gy irradiation (SF2). RESULTS: In our investigation, the extrapolation method proved to be inappropriate for the study of in vitro cellular radiosensitivity due to lack of reproducibility. The results obtained by the clonogenic assay showed that the cell lines studied were radiobiologically heterogeneous with no discrete features of the examined parameters including the repair capacity. CONCLUSION: The results indicate that SCLC tumors per se are not generally candidates for hyperfractionated radiotherapy. PMID- 9212022 TI - Differential influence of TGFbeta1 and TGFbeta3 isoforms on cell cycle kinetics and postirradiation recovery of normal and malignant colorectal epithelial cells. AB - PURPOSE: A clonogenic assay was used to determine the effects of the growth factors TGFbeta1 and TGFbeta3 on the radiation responses of a normal rat epithelial cell line (IEC6) and a human colonic carcinoma epithelial cell line (Widr). METHODS AND MATERIALS: The radiation sensitivity and ability to recover from potentially lethal damage (PLD), of preconfluent monolayer cultures, was assessed in the presence of the growth factors for 24 h prior to, during, and after irradiation. RESULTS: The surviving fractions of both cell lines assessed immediately following irradiation were unaffected by TGFbeta1 or TGFbeta3. However, TGFbeta3 (but not TGFbeta1) significantly reduced the amount of PLD recovery in the Widr cells (but not in the IEC6 cells). This was associated with a reduction in the shoulder region of the survival curve, rather than a change in slope. A comparative analysis of the effects of TGFbetas 1 and 3 on cell cycle events in the two cell lines demonstrated significantly more Widr cells in the S phase, in the presence of TGFbeta3 only, compared to the controls. This remained constant both before and immediately following irradiation. In the IEC6 cell line TGFbeta3 produced an increase in the numbers of G1 phase cells, characteristic of a G1 arrest. CONCLUSION: It seems likely that TGFbeta3-induced radiosensitisation in Widr cells, 6 h after a single dose of irradiation, is related to its effects on cell cycle events such that the failure of these cells to arrest in G1, either before or after irradiation, results in significantly reduced recovery from DNA damage. This, however, may not be the only mechanism by which this growth factor produces this effect. Indeed, it will also be necessary to investigate these effects in in vivo models and to determine the response to fractionated irradiation before the potential therapeutic benefit of both the differential effects observed between the two TGFbeta isoforms and also between the malignant and normal cell lines can be fully assessed. PMID- 9212024 TI - Adaptive modification of treatment planning to minimize the deleterious effects of treatment setup errors. AB - PURPOSE: Using daily setup variation measured from an electronic portal imaging device (EPID), radiation treatment of the individual patient can be adaptively reoptimized during the course of therapy. In this study, daily portal images were retrospectively examined to: (a) determine the number of initial days of portal imaging required to give adequate prediction of the systematic and random setup errors; and (b) explore the potential of using the prediction as feedback to reoptimize the individual treatment part-way through the treatment course. METHODS AND MATERIALS: Daily portal images of 64 cancer patients, whose treatment position was not adjusted during the course of treatment, were obtained from two independent clinics with similar setup procedures. Systematic and random setup errors for each patient were predicted using different numbers of initial portal measurements. The statistical confidence of the predictions was tested to determine the number of daily portal measurements needed to give reasonable predictions. Two treatment processes were simulated to examine the potential opportunity for setup margin reduction and dose escalation. The first process mimicked a conventional treatment. A constant margin was assigned to each treatment field to compensate for the average setup error of the patient population. A treatment dose was then prescribed with reference to a fixed normal tissue tolerance, and then fixed in the entire course of treatment. In the second process, the same treatment fields and prescribed dose were used only for the initial plan and treatment. After several initial days of treatments, the treatment field shape and position were assumed to be adaptively modified using a computer-controlled multileaf collimator (MLC) in light of the predicted systematic and random setup errors. The prescribed dose was then escalated until the same normal tissue tolerance, as determined in the first treatment process, was reached. RESULTS: The systematic setup error and the random setup error were predicted to be within +/-1 mm for the former and +/-0.5 mm for the latter at a > or = 95% confidence level using < or = 9 initial daily portal measurements. In the study, a large number of patients could be treated using a smaller field margin if the adaptive modification process were used. Simulation of the adaptive modification process for prostate treatment demonstrates that additional treatment dose could be safely applied to 64% of patients. CONCLUSION: The adaptive modification process represents a different approach for use of on-line portal images. The portal imaging information from the initial treatments is used as feedback for reoptimization of the treatment plan, rather than adjustment of the treatment setup. Results from the retrospective study show that the treatment of individual patient can be improved with the adaptive modification process. PMID- 9212025 TI - HDR source calibration methods and discrepancies. AB - PURPOSE: To report the calibration discrepancies observed in HDR sources and our methods of calibration of a new source. METHODS AND MATERIALS: 21 sources have been calibrated since March 1991 to February 1996 and discrepancies are reported in comparison to the certificate value. The calibration methodology used for two different systems: the NEL Farmer system and reentrant well chamber (HDR-1000) are described. A lead shield insert developed for use with the well chamber for timer linearity measurement is described. RESULTS: Eleven sources had discrepancy from 0 to 1%, 7 sources had discrepancy from 1 to 5%, and 3 sources had discrepancy from 5 to 10%. The maximum discrepancy observed was -8.04 % for one source. Our procedure of dealing with situations when discrepancy exceeds the range of +/-5% value is discussed. CONCLUSION: (a) The discrepancies that we observed in our study are compared with other users and found to be within the range of values observed. (b) Due to the easy set up procedure, less prone to error in the use of well chamber and with the use of a new lead insert, we conclude in our case that, the well system can be used for monthly calibration of HDR source and timer linearity measurements. (c) Because the Farmer system has a first generation calibration factor, obtained by interpolation of two calibration energies from ADCL, we prefer to use the Farmer system for determining the activity of the new HDR source. PMID- 9212026 TI - Integration of radiotherapy planning systems and radiotherapy treatment equipment: 11 years experience. AB - PURPOSE: We have investigated the requirements, design, implementation, and operation of a computer-controlled medical accelerator with multileaf collimator (MLC), integrated with a radiation treatment-planning system (RTPS), and we report on the performance, benefits, and lessons learned from this experience. METHODS AND MATERIALS: In 1984 the University of Washington installed a computer controlled radiation therapy machine (the Clinical Neutron Therapy System, or CNTS) with a multileaf collimator. Since the beginning of operation the control system computer has been connected by commercially available network hardware and software to three generations of radiation treatment-planning systems. Semiautomated setup and completely computerized check and confirm were incorporated into the system from the beginning of clinical operation in 1984. The system cannot deliver a patient treatment without a computer-prepared treatment plan. RESULTS: The CNTS has been in use for routine patient treatments for over 11 years. The cost of the network connection and software was an insignificant fraction of the facility cost. Operation has been efficient and reliable. Of the 441 machine-related session reschedulings (out of 18,432 sessions total) during the past 9 years, only 20 were due to problems with data transfer between the RTPS and CNTS, associated primarily with two incidents. Close integration with the treatment-planning system allows complex treatments to be delivered. Dramatic evolution of the departmental treatment-planning system has not required any changes or redesign of either the accelerator control system or the network connection. CONCLUSIONS: Our experience shows that a large degree of automation is possible with reasonable effort, by using well-known software and hardware design strategies. The lessons we have learned from this can be carried over into photon therapy now that photon accelerators with MLC facilities are commercially available. PMID- 9212027 TI - Sinusitis: bench to bedside. Current findings, future directions. AB - Sinusitis, an inflammatory disease of the sinus, is one of the most commonly reported diseases in the United States, affecting an estimated 14% of the population. The prevalence of sinusitis is rising. Between 1990 and 1992, persons with sinusitis reported approximately 73 million restricted activity days-an increase from the 50 million restricted activity days reported between 1986 and 1988. Because critical questions remain unanswered about its cause, pathophysiology, and optimal treatment, sinusitis continues to generate significant health care costs and affects the quality of life of a large segment of the U.S. population. To identify critical directions for research on sinus disease, the American Academy of Allergy, Asthma and Immunology and the American Academy of Otolaryngology-Head and Neck Surgery Foundation, Inc., convened a meeting in January 1996 in collaboration with the National Institutes of Allergy and Infectious Disease. This document summarizes the proceedings of that meeting and presents what is intended to be the background for future investigation of the many unanswered questions related to sinusitis. PMID- 9212028 TI - Sinusitis: bench to bedside. Current findings, future directions. AB - Sinusitis, an inflammatory disease of the sinus, is one of the most commonly reported diseases in the United States, affecting an estimated 14% of the population. The prevalence of sinusitis is rising. Between 1990 and 1992, persons with sinusitis reported approximately 73 million restricted activity days--an increase from the 50 million restricted activity days reported between 1986 and 1988. Because critical questions remain unanswered about its cause, pathophysiology, and optimal treatment, sinusitis continues to generate significant health care costs and affects the quality of life of a large segment of the U.S. population. To identify critical directions for research on sinus disease, the American Academy of Allergy, Asthma and Immunology and the American Academy of Otolaryngology-Head and Neck Surgery Foundation, Inc., convened a meeting in January 1996 in collaboration with the National Institutes of Allergy and Infectious Disease. This document summarizes the proceedings of that meeting and presents what is intended to be the background for future investigation of the many unanswered questions related to sinusitis. PMID- 9212029 TI - Maternal intrauterine infection, cytokines, and brain damage in the preterm newborn. AB - To evaluate the hypothesis that the proinflammatory cytokines IL-1, IL-6, and tumor necrosis factor-alpha might be the link between prenatal intrauterine infection (IUI) and neonatal brain damage, the authors review the relevant epidemiologic and cytokine literature. Maternal IUI appears to increase the risk of preterm delivery, which in turn is associated with an increased risk of intraventricular hemorrhage, neonatal white matter damage, and subsequent cerebral palsy. IL-1, IL-6, and TNF-alpha have been found associated with IUI, preterm birth, neonatal infections. and neonatal brain damage. Unifying models not only postulate the presence of cytokines in the three relevant maternal/fetal compartments (uterus, fetal circulation, and fetal brain) and the ability of the cytokines to cross boundaries (placenta and blood-brain barrier) between these compartments, but also postulate how proinflammatory cytokines might lead to IVH and neonatal white matter damage during prenatal maternal infection. Interrupting the proinflammatory cytokine cascade might prevent later disability in those born near the end of the second trimester. PMID- 9212030 TI - The life span of erythrocytes transfused to preterm infants. AB - This study was made to determine the life span of adult red cells transfused to early preterm infants. Nineteen very preterm infants (birth weight, 878.7 +/- 221 g; gestational age, 26.8 +/- 1.5 wk at birth) were sampled weekly after their last blood transfusion to determine the level (%) of fetal Hb in their circulation. Two microliters of blood were subjected to reverse phase HPLC to separate the alpha, beta, and gamma globin components of their Hbs. The percent of fetal Hb (HbF) was calculated as gamma/gamma + beta x 100. The life span of the adult erythrocytes transfused was defined as the time interval between the transfusion and when the percentage of HbF in the recipient's circulation returns to the HbF levels that exist in the infant's autologous red cells (the maximum post transfusion HbF level). Twelve of the 19 infants were followed until their autologous HbF levels were reached. Their mean adult red blood cell life span was 56.4 +/- 7 d (range: 46-68 d). The results obtained in this study imply that the number of days after a transfusion at which half the cells infused remain in the circulation in a preterm infant is about 30 d. PMID- 9212031 TI - Right ventricular infundibular beta-adrenoceptor complex in tetralogy of Fallot patients. AB - Patients with tetralogy of Fallot may have episodes of paroxysmal hypoxic spells ("tet spells") or could be asymptomatic. In patients who have these episodes, treatment with a beta-adrenoceptor (betaAR) blocking agent can often ameliorate or attenuate the severity of the symptoms. Additionally, excitement, crying, and situations associated with increased sympathetic activity could provoke the occurrence of these hypoxic spells. We hypothesized that altered myocardial betaAR function may contribute to the development of paroxysmal hypoxic spells in the symptomatic tetralogy patient. Surgically excised right ventricular infundibular myocardial specimens from symptomatic (patients with spells) and asymptomatic patients were used to determine total beta1 and beta2 betaAR density and betaAR adenylyl cyclase activity. Symptomatic patients had a significantly greater number of total betaAR. The relative proportion of beta1 and beta2 receptors was comparable in both patient groups. betaAR-stimulated adenylyl cyclase activity was found to be more enhanced in the symptomatic patient group. Our results indicate that infundibular betaARs may play a role in the development of paroxysmal hypoxic spells. PMID- 9212032 TI - Modest hypothermia provides partial neuroprotection when used for immediate resuscitation after brain ischemia. AB - Intraischemic reduction in temperature of 2-3 degrees C (modest hypothermia) has been demonstrated to provide partial neuroprotection in neonatal animals. This investigation determined if modest hypothermia initiated immediately after brain ischemia provides neuroprotection. Piglets were studied with rectal temperature maintained during the 1st h after 15 min of brain ischemia at either 38.3 +/- 0.3 degrees C (normothermia, n = 11) or at 35.8 +/- 0.5 degrees C (modest hypothermia, n = 11). The severity of brain ischemia was similar between groups as indicated by equivalent reduction in mean blood pressure (90 +/- 15 to 24 +/- 3 versus 92 +/- 13 to 26 +/- 3 mm Hg), and changes in cerebral metabolites and intracellular pH (pH(i)) measured by magnetic resonance spectroscopy (beta nucleoside triphosphate = 44 +/- 9 versus 42 +/- 18% of control, control = 100%, pH(i): 6.25+/- .15 versus 6.24 +/- 0.22 for normothermic and modestly hypothermic groups, respectively). In the first 90 min after ischemia, there were no differences between groups in the duration and extent of brain acidosis, and relative concentrations of phosphorylated metabolites. Categorical assessment of neurobehavior was evaluated at 72 h postischemia (n = 16), or earlier if an animal's condition deteriorated (n = 6). Postischemic hypothermia was associated with less severe stages of encephalopathy compared with normothermia (p = 0.05). Histologic neuronal injury was assessed categorically in 16 brain regions, and postischemic hypothermia resulted in less neuronal injury in temporal (p = 0.024) and occipital (p = 0.044) cortex at 10 mm beneath the cortical surface, and in the basal ganglia (p = 0.038) compared with that in normothermia. Modest hypothermia for 1 h immediately after brain ischemia provides partial neuroprotection and may represent an adjunct to resuscitative strategies. PMID- 9212033 TI - Effect of carbon dioxide on cerebral metabolism during hypoxia-ischemia in the immature rat. AB - We previously have demonstrated that hypocapnia aggravates and hypercapnia protects the immature rat from hypoxic-ischemic brain damage. To ascertain cerebral blood flow (CBF) and metabolic correlates, 7-d postnatal rats were subjected to hypoxia-ischemia during which they were rendered either hypo-(3.5 kPa), normo- (5.1 kPa), or hypercapnic (7.3 kPa) by the inhalation of either 0, 3, or 6% CO2, 8% O2, balance N2. CBF during hypoxia-ischemia was better preserved in the normo- and hypercapnic rat pups; these animals also exhibited a stimulation of cerebral glucose utilization. Brain glucose concentrations were higher and lactate lower in the normo- and hypercapnic animals, indicating that glucose was consumed oxidatively in these groups rather than by anaerobic glycolysis, as apparently occurred in the hypocapnic animals. ATP and phosphocreatine were better preserved in the normo- and hypercapnic rats compared with the hypocapnic animals. Cerebrospinal fluid glutamate, as a reflection of the brain extracellular fluid concentration, was lowest in the hypercapnic rats at 2 h of hypoxia-ischemia. The data indicate that during hypoxia-ischemia in the immature rat, CBF is better preserved during normo- and hypercapnia; the greater oxygen delivery promotes cerebral glucose utilization and oxidative metabolism for optimal maintenance of tissue high energy phosphate reserves. An inhibition of glutamate secretion into the synaptic cleft and its attenuation of N-methyl-D aspartate receptor activation would further protect the hypercapnic animal from hypoxic-ischemic brain damage. PMID- 9212034 TI - Long-term impairment in the neurochemical activity of the sympathoadrenal system after neonatal hypoxia in the rat. AB - The study evaluates the long-term effect of neonatal hypoxia on the neurochemical activity of the sympathoadrenal system in the rat. One-day-old male pups were exposed to hypoxia (10% O2) for 6 d and thereafter reared under normoxia. Neonatal hypoxia reduced the body weight of 3- and 8-wk-old rats and did not change the blood pressure at 6 wk of age. In sympathetic ganglia, the content and/or turnover rates of norepinephrine were reduced in neonatal-hypoxic rats of 3 and 8 wk of age, but the content and turnover rates of dopamine were unaltered. The effect was not dependent on the type of ganglion. In the superior cervical ganglion, neonatal hypoxia had a selective effect on the type of catecholamine (dopamine versus norepinephrine), thus suggesting a selective-altered maturation of noradrenergic neurons, but presumably not of the dopaminergic small, intensely fluorescent cells. A long-term deficiency in adrenal activity was the consequence of neonatal hypoxia, as shown by the decrease in the content and turnover rate of dopamine. Neonatal hypoxia elicited a long-term decrease in the content and turnover rates of norepinephrine in heart and lungs but failed to induce a significant effect in kidneys. However, this effect was not tissue-specific. Data provide evidence that a hypoxic episode occurring during a critical period of development in the rat induces a long lasting decrease in the neurochemical activity of the sympathoadrenal system. These results are discussed in terms of their implications for human pathology. PMID- 9212035 TI - Newborn cerebrovascular responses after first trimester moderate maternal ethanol exposure in sheep. AB - Fetal alcohol syndrome is one of the leading causes of mental retardation in the United States, but the pathogenesis of the associated brain damage is unknown. We tested the hypothesis that neonatal cerebrovascular responses to CO2 and/or hypoxia may be altered by moderate chronic maternal ethanol exposure early in gestation. We studied 26 newborn lambs (1-4 d old). Their mothers had received daily i.v. infusions of either ethanol (1 g/kg; ethanol concentration = 167 +/- 3 mg/dL; mean +/- SEM) or a similar volume of saline for 3 wk during the first trimester. In nine lambs, we studied cerebral responses to CO2 (saline, n = 4; ethanol, n = 5) and in 17 lambs, cerebral responses to hypoxia (saline, n = 7; ethanol, n = 10). Cerebrovascular responses to CO2 were not different between the groups. However, the cerebral vasodilatory response to hypoxemia was significantly attenuated in the ethanol lambs, such that cerebral O2 delivery was not maintained. During severe hypoxia (arterial PO2 = 30 mm Hg), cerebral blood flow increased 106 +/- 23% (mean +/- SEM) above baseline in the saline-treated group, but increased only 32 +/- 15% above baseline in the ethanol-treated group (p < 0.02). Similarly, cerebrovascular resistance in the saline group decreased 52 +/- 6% from baseline, but decreased only 16 +/- 11% in the ethanol group (p < 0.02). We conclude that moderate maternal ethanol infusion early in pregnancy attenuates neonatal hypoxic, but not CO2, cerebrovascular responsivity. PMID- 9212036 TI - Adenosine modulates inspiratory neurons and the respiratory pattern in the brainstem of neonatal rats. AB - The role of adenosine in the modulation of respiration-related neurons was examined using an in vitro brainstem-spinal cord preparation from neonatal rats (0-4 d old). Respiratory activity was recorded from the C4 or C5 ventral roots by suction electrodes and from inspiratory related neurons (I neurons) in the rostral ventrolateral medulla by microelectrodes. The following substances were added to the preparation superfusate, and their effect was evaluated: the adenosine A1 receptor agonist N6-(2-phenylisopropyl)adenosine, R(-)isomer (R PIA), the adenosine uptake blocker dipyridamole, the adenosine receptor antagonist theophylline, and the specific A1 receptor antagonist 8-cyclopentyl 1,3-dipropylxanthine (DPCPX). R-PIA and dipyridamole decreased the activity of I neurons and the C4 respiratory burst rate. Furthermore, these compounds induced a significantly more irregular respiratory rate in three-quarters of preparations from the youngest animals (<48 h old) compared with that of controls. Theophylline or DPCPX reversed the effects of both R-PIA and dipyridamole on respiratory rate, regularity of respiratory rate, inspiratory time, amplitude, and intra-burst frequency of I neurons. Thus, adenosine depresses both the I neurons in the rostral ventrolateral medulla and the respiratory motor output. This depression of I neurons and respiratory rate can be abolished by theophylline primarily through a blockade of medullary adenosine A1 receptors. An age-dependent correlation of the effects of R-PIA and dipyridamole, with a more pronounced decrease in respiratory activity in preparations from younger animals, indicates that adenosinergic modulation of medullary respiration-related neurons changes during the first days of postnatal life. PMID- 9212037 TI - Late glial swelling after acute cerebral hypoxia-ischemia in the neonatal rat: a combined magnetic resonance and histochemical study. AB - Secondary brain damage after transient cerebral hypoxia-ischemia (HI) is caused by a cascade of cellular events. In this study, complementary methods of magnetic resonance imaging and histochemistry were used to investigate the formation of cytotoxic and vasogenic edema during secondary brain damage induced by transient HI in 7-d-old rats. To elicit injury, 21 rats underwent right common carotid artery ligation followed by 1.5 h of 8% O2 exposure. Sequential apparent diffusion coefficient (ADC) and transversal relaxation time (T2) weighted magnetic resonance imaging were recorded for up to 3 d in 13 7-d-old rats. Eight animals were killed at various intervals between the end of HI and 21 h of recovery to perform histochemical assays using neuronal and astrocytic markers. Changes of the ADC revealed a biphasic function for the evolution of cytotoxic edema during the recovery period. At the end of HI, the ADC in the ipsilateral cortex was significantly decreased. Upon reoxygenation, it returned transiently to normal followed by a secondary, although less pronounced, decline after 8-48 h. After this, the ADC rose steadily. From 8 h of recovery, the proportion of vasogenic edema steadily increased as indicated by the T2 prolongation. At 21 h, the majority of glial cells showed immunoreactivity for glial fibrillary acidic protein and were of larger size, whereas the neurons were apoptotic. These results indicate that the delayed cerebral injury is accompanied by late glial swelling in conjunction with an enlarged interstitial space due to cell damage. PMID- 9212038 TI - Rat diaphragm oxidative capacity, antioxidant enzymes, and fatigue: newborn versus adult. AB - Little is known about the antioxidant capacity and oxidant-generating potential of newborn muscle, or how these properties compare with the adult and relate to fatigue resistance. We determined the 1) antioxidant enzyme activities [superoxide dismutase (SOD), catalase, glutathione peroxidase], 2) glutathione content, 3) oxidative capacity [indexed by succinic dehydrogenase activity], 4) extracellular cytochrome c reduction, and 5) efficacy of exogenously administered SOD in ameliorating fatigue in vitro of newborn and adult diaphragm (DIA). Newborn and adult DIA SOD activities were not different, whereas newborn catalase activity was greater, and newborn glutathione peroxidase activity and glutathione content less than adult DIA. Succinic dehydrogenase activity was approximately 2 fold greater in the adult compared with the neonate. Repetitive contractions led to a significant decline in newborn and adult DIA force; this decline was greater in the adult (78 +/- 4% decrement in force at 2 min) compared with newborn DIA (28 +/- 8% decrement in force at 2 min). Extracellular cytochrome c reduction was greater in adult as compared with newborn DIA during fatiguing contractions. Exogenous SOD attenuated fatigue in the adult, but had no effect on newborn DIA. We conclude that the oxidative capacity of the adult DIA is greater than that of the newborn and not matched by a concomitant increase in SOD activity. Our data suggest that the increased oxidative capacity relative to SOD activity in adult DIA may lead to oxidative stress and an enhanced susceptibility to fatigue. PMID- 9212039 TI - The use of an automated microsampling system for the characterization of growth hormone pulsatility in newborn babies. AB - To overcome the difficulties of studying hormone pulsatility in the newborn, we have developed an automated microsampling system that permits the measurement of hormones in small prediluted samples of blood (40 microL) taken at 10-min intervals over 12 h. The system has been validated in adult volunteers, and the error attributable to the dilution was <4%. Using this method in 10 preterm babies, we have been able to describe pulsatile changes in GH and have demonstrated a clear postprandial elevation in GH levels peaking 60 min after a feed. Fourier transform analysis indicated a pulse periodicity of 180 min in babies who were appropriate for gestational age (n = 6), but faster, co-dominant pulse periodicities of 90-100 and 140 min in babies who were small for gestational age (weight and length below the 10th centile) (n = 4). There was no significant difference between mean, peak, and baseline GH levels between the two groups. PMID- 9212040 TI - Major circadian variations of glucose homeostasis in a patient with Rabson Mendenhall syndrome and primary insulin resistance due to a mutation (Cys284- >Tyr) in the insulin receptor alpha-subunit. AB - We have performed clinical, in vitro biochemical, and genetic studies of a patient with severe insulin resistance, considerable growth restriction, and Rabson-Mendenhall syndrome (patient RM-3). The blood IGF-I level was undetectable in this patient, although the GH level was moderately decreased. During the postprandial period, glycemia, ketonuria, and plasma glucagon were very elevated despite high doses of exogenous insulin (glucose levels up to 30 mmol/L). In the postabsorptive state, blood glucose was normalized with small amounts of insulin; ketonuria, and glucagon levels were reduced but remained supranormal. Erythrocytes and cultured skin fibroblasts from the patient displayed a decrease in cell surface insulin receptors (IRs). The ability of physiologic concentrations of insulin to stimulate metabolic processes was altered in patient fibroblasts. Analysis of the IR gene by denaturing gradient gel electrophoresis and direct sequencing showed a homozygous missense mutation in exon 3, replacing Cys284 by Tyr in the alpha-subunit. In conclusion, marked primary insulin resistance was evidenced in patient cells as a result of a structural alteration in the IR alpha-subunit. The in vitro studies could not account alone for the in vivo metabolic alterations because glucose homeostasis varied considerably during the day in the patient. PMID- 9212041 TI - Small intestinal disaccharidase activity and ileal villus height are increased in piglets consuming formula containing recombinant human insulin-like growth factor I. AB - The effect of orally administered IGF-I on intestinal development was assessed in piglets. Cesarean-derived, colostrum-deprived piglets received formula alone or formula containing 65 nM (500 microg/L) of recombinant human IGF-I. IGF-I intake averaged 200 microg/kg/d. On d 7 and 14 postpartum, piglets were killed, organs were removed and weighed, and tissue and blood samples were collected. The small intestine was divided into 13 segments that were weighed and measured. A sample of each segment was fixed in formalin, and the mucosa was scraped for enzyme analyses. Food intake, body and organ weights, intestinal weight, length, protein, DNA and RNA content did not differ between the treatment groups. Serum IGF-I, IGF-II, and IGF-binding protein profiles and tissue IGF-binding protein mRNA expression were also comparable between the treatment groups. In contrast, intestinal enzymes and villus height were increased by oral IGF-I. Lactase was approximately 2-fold higher (p < or = 0.05) in the jejunum and proximal ileum, and sucrase was approximately 50% higher (p < or = 0.05) in the jejunum of IGF-I treated animals than in controls. Villus height in the terminal ileum was approximately 50% greater in IGF-I-treated animals than in controls (p = 0.03). In conclusion, orally administered IGF-I at 200 microg/kg did not affect whole body or organ growth or serum IGF-I concentrations; however, intestinal disaccharidase activity and ileal villus growth were responsive to orally administered IGF-I, supporting a potential role for milk-borne IGF-I in neonatal intestinal development. PMID- 9212042 TI - L-thyroxine treatment of preterm newborns: clinical and endocrine effects. AB - Preterm newborns have low serum thyroxine (T4) levels compared with late gestational fetuses. Low thyroid hormone levels are associated with increased severity of neonatal illness and neurodevelopmental dysfunction. We assessed the endocrine and clinical effects of increasing serum T4 levels in preterm newborns with a gestational age <31 wk. Forty newborns were randomized in a double blind protocol: 20 infants received a daily dose of 20 microg/kg L-T4 for 2 wk, whereas 20 control infants received saline. Serum concentrations of T4, triiodothyronine (T3), reverse T3 (rT3), thyroglobulin (TG), and TSH were measured weekly as well as serum levels of GH, prolactin, and IGF-I. After 2 wk, a TSH-releasing hormone (TRH) test was performed. Neonatal illness and outcome was evaluated by noting heart rate, oxygen requirement, duration of ventilation, development of chronic lung disease, oral fluid intake, and weight gain; a Bayley score was done at the corrected age of 7 mo. L-T4 administration induced a marked increase in serum T4 without apparent change in T3 levels, whereas the postnatal decline in serum rT3 was more gradual. L-T4 treatment was associated with a decrease in serum TG and TSH levels. TRH injection induced a definite rise in serum TSH and T3 in controls, but not in L-T4 treated newborns. Neither L-T4 treatment, nor TRH administration appeared to alter circulating levels of prolactin, GH, or IGF-I. In contrast to the pronounced endocrine effects, no clinical effects of L-T4 administration were detected. PMID- 9212043 TI - A critical period for the role of thyroid hormone in development of renal alpha adrenergic receptors. AB - Adrenergic input influences renal cell replication/differentiation and the development of excretory function. Kidney cells make adrenoceptors before the arrival of the majority of nerve terminals, and the current study examines whether thyroid hormone plays a role in receptor development. Propylthiouracil (PTU) was given to pregnant and neonatal rats from gestational d 17 through postnatal d 5, a treatment that obtunds thyroid hormone levels throughout the first 2-3 wk postpartum. The PTU group showed significant deficits in the number of alpha1-receptors, and values resolved to normal in parallel with hormone level recovery. The effects were not secondary to alterations in cell differentiation or growth. as the period of receptor abnormalities did not correspond to that of growth inhibition. Similarly, the effects were selective for the alpha1-receptor, as no comparable effects were seen for total membrane protein or for alpha2 receptors. The role of thyroid hormone in alpha1-receptor ontogeny involved a critical developmental window; later in development neither treatment with PTU nor with large doses of thyroid hormone had any impact on alpha1-receptors. Studies of mRNAs encoding the alpha1-receptor subtypes indicated that hypothyroidism targets the alpha1a-subtype, which has been implicated in the transduction of neurotrophic signals; alpha1a-receptor mRNA also showed the largest proportional developmental increase compared with those encoding other alpha1-subtypes. Accordingly, thyroid hormone is likely to set the stage for the subsequent trophic control of renal development by neural input, and hypothyroidism during this critical window can be expected to result in abnormal renal functional development and increased perinatal risk. PMID- 9212044 TI - Cardiovascular defects among the progeny of mouse phenylketonuria females. AB - In a genetic mouse model of human phenylketonuria we have examined the offspring of hyperphenylalaninemic mothers for the presence of cardiovascular defects, an important feature of the pathology of the human maternal phenylketonuria syndrome. Beginning at 14.5 d after conception (75% through gestation), a variety of cardiovascular defects became apparent among the progeny of the hyperphenylalaninemic females. These defects ranged from mild to serious and correlated with the maternal but not the fetal Pah genotype. Nearly all of the defects were vascular, however, whereas the most reported in humans so far have been cardiac. The predisposing biochemical condition in this mouse disease model seems to be the same as in the human disease; elevated maternal blood phenylalanine levels concentrated across the placental barrier to produce a teratogenic developmental environment. This model for congenital cardiovascular defects should enhance two related areas of research. 1) It should allow a more thorough investigation of the relationship between maternal diet and maternal phenylketonuria birth defects, and 2) it should provide an experimental tool to gain insight into the normal process of cardiovascular development. PMID- 9212045 TI - The effect of carnitine on ketogenesis in perfused livers from juvenile visceral steatosis mice with systemic carnitine deficiency. AB - Juvenile visceral steatosis (JVS) mice have been reported to have systemic carnitine deficiency, and the carnitine concentration in the liver of JVS mice was markedly lower than that of controls (11.6 +/- 2.6 versus 393.5 +/- 56.4 nmol/g of wet liver). To evaluate the role of carnitine in mitochondrial beta oxidation in liver, we examined the effects of carnitine on ketogenesis in perfused liver from control and JVS mice. In control mice, ketogenesis was increased by the infusion of 0.3 mM oleate, but not by L-carnitine. In contrast, although ketogenesis in JVS mice was not increased by the infusion of oleate, it was increased 2.5-fold by the addition of 1000 microM L-carnitine. Addition of 50, 100, and 200 microM L-carnitine increased ketogenesis in a dose-dependent manner. The infusion of 0.3 mM octanoate or butyrate increased ketogenesis in a carnitine-independent fashion in both control and JVS mice. These findings suggest that endogenous long chain fatty acids from accumulated triglycerides may be used as substrates in the presence of carnitine in JVS mice. The relationship between ketogenesis and free carnitine concentration was examined in livers from JVS mice. Ketogenesis increased as free carnitine levels increased until concentrations exceeded about 100 nmol/g of wet liver (340 microM). The free carnitine concentration required for half-maximal ketone body production in liver of JVS mice was 45 microM (13 nmol/g of wet liver), which corresponds to a K(m) value of carnitine palmitoyltransferase I. We conclude that carnitine is a rate limiting factor for beta-oxidation in liver only when the carnitine level in liver is very low. PMID- 9212046 TI - Neonatal nutritional carnitine deficiency: a piglet model. AB - The clinical significance of nutritional carnitine deficiency remains controversial. To investigate this condition under controlled conditions, an animal model was developed using the parenterally alimented, carnitine-deprived newborn piglet. Forty-five piglets received total parenteral nutrition for 2-3 wk that was either carnitine-free or supplemented with 100-400 mg/L L-carnitine. Blood and a muscle biopsy were taken at the initial surgery. Carnitine balance studies were performed at 11-14 d of age. Blood, liver, heart, and skeletal muscle were taken at sacrifice for analysis of carnitine, electron microscopy, and oxidation studies. Carnitine-deprived piglets were in negative carnitine balance and had lower blood, urine, and tissue levels of carnitine than carnitine supplemented animals. There was a positive correlation between excretion and plasma concentrations of free carnitine with an apparent renal threshold between 15 and 35 micromol/L. Plasma levels were correlated with liver and heart, but not muscle, concentrations of total acid-soluble carnitine. Carnitine-deprived piglets had evidence of lipid deposition in liver and skeletal muscle and tended to have a higher incidence of muscle weakness and cardiac failure. Basal rates of oxidation of [14C]palmitate to 14CO2 and 14C-acid-soluble products were lower in liver homogenates from carnitine-deprived piglets than in those from carnitine supplemented animals and increased in a dose-dependent fashion with the addition of L-carnitine (0, 50, and 500 micromol/L) in vitro. In summary, carnitine deprivation in the neonatal piglet resulted in low carnitine status and morphologic/functional disturbances compatible with carnitine deficiency. The described animal model appears to be suitable for the investigation of neonatal nutritional carnitine deficiency. PMID- 9212047 TI - HLA-A2402-restricted and tumor-specific cytotoxic T lymphocytes from tumor infiltrating lymphocytes of a child with Wilms' tumor. AB - The existence of specific immunity against pediatric tumors is not well studied. We report here the CD3+CD4-CD8+ cytotoxic T lymphocyte (CTL) line established from tumor-infiltrating lymphocytes (TIL) of a 3-mo-old child with Wilms' tumor. This CD3+CD4-CD8+ CTL line showed cytotoxicity against the HLA-A2402+ tumor cells including the autologous tumor, adenocarcinomas from various organs (colon, stomach, lung, and ovary), and an esophageal squamous cell carcinoma. No other cell lines examined, including HLA-A2402- tumors, K562 cells, or HLA-A2402+ normal cells were lysed by this CTL line. This CTL line recognized an HLA-A2402- ovarian adenocarcinoma transfected with HLA-A2402 cDNA. These results suggest the existence of HLA-A2402-restricted and tumor-specific CTL at the tumor site of a child with Wilms' tumor. PMID- 9212048 TI - The administration of complement component C9 enhances the survival of neonatal rats with Escherichia coli sepsis. AB - To determine the significance of neonatal C9 deficiency, an animal model was developed in the rat. By rocket immunoelectrophoresis, the concentration of C9 in pooled adult rat serum was 224 +/- 7.2 microg/mL. In contrast, the concentration of C9 in pooled serum from 1-d-old rats was only 43 +/- 3.8 microg/mL and increased during the first 3 wk of life to 170 +/- 20 microg/mL. Similarly, the capacities of neonatal rat serum to kill two pathogenic strains of Escherichia coli and to lyse sensitized sheep erythrocytes were diminished compared with adult serum but increased during the first 3 wk of life. Supplemental human C9 significantly enhanced the bactericidal and hemolytic activity of neonatal rat serum. The capacity of neonatal rats to survive after the intrapulmonary injection of E. coli was positively correlated with the serum C9 concentration, bactericidal activity, and hemolytic activity. In 2-d-old rats infected with E. coli, the intraperitoneal administration of human C9 significantly enhanced survival and also enhanced the protective effect of intraperitoneal human IgG antibodies. The data indicate that C9 deficiency predisposed neonatal rats to invasion by E. coli. The neonatal rat appears to be a suitable model with which to investigate the significance of C9 deficiency. PMID- 9212049 TI - High-mobility group (HMG) protein HMG-1 and TATA-binding protein-associated factor TAF(II)30 affect estrogen receptor-mediated transcriptional activation. AB - The estrogen receptor (ER) belongs to a family of ligand-inducible nuclear receptors that exert their effects by binding to cis-acting DNA elements in the regulatory region of target genes. The detailed mechanisms by which ER interacts with the estrogen response element (ERE) and affects transcription still remain to be elucidated. To study the ER-ERE interaction and transcription initiation, we employed purified recombinant ER expressed in both the baculovirus-Sf9 and his tagged bacterial systems. The effect of high-mobility group (HMG) protein HMG-1 and purified recombinant TATA-binding protein-associated factor TAF(II)30 on ER ERE binding and transcription initiation were assessed by electrophoretic mobility shift assay and in vitro transcription from an ERE-containing template (pERE2LovTATA), respectively. We find that purified, recombinant ER fails to bind to ERE in spite of high ligand-binding activity and electrophoretic and immunological properties identical to ER in MCF-7 breast cancer cells. HMG-1 interacts with ER and promotes ER-ERE binding in a concentration- and time dependent manner. The effectiveness of HMG-1 to stimulate ER-ERE binding in the electrophoretic mobility shift assay depends on the sequence flanking the ERE consensus as well as the position of the latter in the oligonucleotide. We find that TAF(II)30 has no effect on ER-ERE binding either alone or in combination with ER and HMG-1. Although HMG-1 promotes ER-ERE binding, it fails to stimulate transcription initiation either in the presence or absence of hormone. In contrast, TAF(II)30, while not affecting ER-ERE binding, stimulates transcription initiation 20-fold in the presence of HMG-1. These results indicate that HMG-1 and TAF(II)30 act in sequence, the former acting to promote ER-ERE binding followed by the latter to stimulate transcription initiation. PMID- 9212050 TI - Dominant negative and cooperative effects of mutant forms of prolactin receptor. AB - In addition to a long form of 591 amino acids (aa), two other forms of PRL receptor (PRLR), differing in the length of their cytoplasmic domains, have been identified in the rat. The Nb2 form, lacking 198 aa in the cytoplasmic domain, is able to transmit a lactogenic signal similar to the long form, whereas the short form of 291 aa is inactive. The ability of PRL to activate the promoter of the beta-casein gene or the lactogenic hormone responsive element fused to the luciferase reporter was assessed in Chinese hamster ovary cells or 293 fibroblasts transiently transfected with PRLR cDNAs. The function of the short form was examined after cotransfection of both the long and short forms. These results clearly show that the short form acts as a dominant negative inhibitor through the formation of inactive heterodimers, resulting in an inhibition of Janus kinase 2 (JAK2) activation. The present study also investigates the possible participation of cytoplasmic receptors in the signal transduction pathway, using cotransfection experiments and a new approach that selectively determines the contribution of cytoplasmic receptors in the process of signal transduction. We cotransfected Chinese hamster ovary cells with two cDNA constructs: a cytoplasmic (soluble) form of the receptor with a deleted signal peptide (delta-19), which is unable to bind PRL, and a functionally inactive receptor mutant (lacking box 1), which is anchored in the plasma membrane and able to bind PRL. This approach has allowed us to show that delta-19, lacking expression at the plasma membrane, can transduce the hormonal message, at least to a limited extent (up to 30% of wild type efficiency), providing that association/activation occurs with a PRL-PRLR complex initiated at the cell surface level; box 1 of the cytoplasmic form is necessary to rescue this partial transcriptional activity of the inactive mutant. This partial recovery is also parallel to the partial activation of JAK2, indicating that the signal transduction pathway implicated JAK2. Our results provide evidence that heterodimerization of receptors can be implicated either in the positive or in negative activation of gene transcription. PMID- 9212051 TI - Identification of a functional androgen-response element in the exon 1-coding sequence of the cystatin-related protein gene crp2. AB - Two hormone-responsive segments, one in the region of the promoter and one in intron 1, are identified in two homologous androgen-regulated and differentially expressed rat genes encoding the cystatin-related proteins (CRPs). Footprint analysis with the androgen receptor (AR) DNA-binding domain on the promoter containing fragments reveals an AR-binding site downstream of the transcription start point in the crp2 gene (ARBSd/crp2, +40/+63). It displays an androgen response element-like sequence motif 5'-AGAAGAaaaTGTACA-3' and overlaps with the ATG translation start codon. A double-stranded oligonucleotide containing this sequence forms a DNA-protein complex with the full-length AR synthesized by vaccinia, as seen in band shift assays. Additional AR-binding sites, ARBSu/crp1 and ARBSu/crp2, occur 5' upstream of the transcription start point and are located at an identical position (-142/ -120) in crp1 and crp2. The AR affinity for these two slightly different sequence motifs is relatively weak. The biological function of all three AR-binding sites as transcription control elements has been studied. The ARBSd/crp2 element clearly shows androgen-response element characteristics. The contribution of the common upstream element to the androgen-dependent control of reporter gene transcription is less clear. The transcription of a reporter gene construct containing the crp2 footprint fragment crp2F (-273/+88) is hormonally regulated as determined by transfection into the human breast cancer cell line T-47D. Androgens, but also glucocorticoids, efficiently stimulate steroid-dependent transcription of the chloramphenicol acetyltransferase gene. Mutation of the 5'-TGTACA-3' sequence in ARBSd/crp2 destroys the AR binding and abolishes the androgen-dependent synthesis of chloramphenicol acetyltransferase. A large fragment derived from intron 1 of the crp1 and crp2 gene can also provide the androgen-dependent transcription of chimeric constructs in T-47D cells. However, the induction measured is less than the one observed with crp2F (-273/+88), and this activity seems to reside in several subfragments that each display a low but consistent androgen responsiveness. PMID- 9212052 TI - Activin A induction of cell-cycle arrest involves modulation of cyclin D2 and p21CIP1/WAF1 in plasmacytic cells. AB - Activins, members of the transforming growth factor-beta family, have been implicated in the regulation of growth and differentiation of various types of cells. We have recently found that activin A induces apoptotic cell death of plasmacytic cells including B cell hybridoma cells and myeloma cells. In the present study, we demonstrated that activin A caused cell-cycle arrest in the G1 phase before appearance of apoptotic cells in mouse B cell hybridoma cells. Phosphorylation of retinoblastoma protein (Rb) and in vitro Rb kinase activity of cyclin-dependent kinase (CDK)4 was inhibited in activin A-treated cells. Analysis of expression of genes regulating Rb phosphorylation revealed that activin A suppressed cyclin D2, the sole D-type cyclin gene expressed in the hybridoma cells, and activated p21CIP1/WAF1 but had no effect on expression of cyclin dependent kinases (CDK2, CDK4, CDK6) and other CDK inhibitors (p27KIP1, p16INK4a, p15INK4b). Modulation of cyclin D2 and p21CIP1/WAF1 expression resulted in a decrease in level of cyclin D2-CDK4 complex and an increase in level of CDK4 complexed with p21CIP1/WAF1. Moreover, overexpression of cyclin D2 partially abrogated inhibition of Rb phosphorylation and G1 arrest in the hybridoma cells. PMID- 9212053 TI - Increased expression of Gs(alpha) enhances activation of the adenylyl cyclase signal transduction cascade. AB - Expression of the stimulatory G protein, G(S)alpha, can vary over a 3-fold range in human tissues and in rodent central nervous system. In fact, the offspring of alcoholics have higher levels of G(S)alpha expression in certain tissues compared with the offspring of nonalcoholics. The aim of this research was to test the hypothesis that a causal relationship exists between the level of expression of G(S)alpha and induction of the adenylyl cyclase (AC) cascade. The methodology employed transient transfection of HEK 293 cells with a cDNA for the 52-kDa form of G(S)alpha under regulation by inducible metallothionein promoters. Transfectants were exposed to varying concentrations (0-125 microM) of zinc sulfate that produced a 3-fold range of membrane G(S)alpha expression. The range of G(S)alpha expression produced was found to mimic a physiologically relevant spectrum of G(S)alpha expression in membranes derived from human tissues and rat brain. It was observed that induction of G(S)alpha expression increased constitutive as well as stimulated cAMP accumulation. Moreover, induction of G(S)alpha expression increased events distal to the accumulation of cAMP including the phosphorylation of the transcription factor, cAMP response element binding protein and transcriptional activation of cAMP-dependent reporter genes. In summary, these studies show that the amount of G(S)alpha expression has a marked impact on the level of activity of the AC cascade from the membrane through to the nucleus. It is hypothesized that individuals who differ in G(S)alpha expression may also differ in the expression of certain cAMP-dependent genes. PMID- 9212054 TI - Activation in vitro of somatostatin receptor subtypes 2, 3, or 4 stimulates protein tyrosine phosphatase activity in membranes from transfected Ras transformed NIH 3T3 cells: coexpression with catalytically inactive SHP-2 blocks responsiveness. AB - Somatostatin receptors (sstr) subtypes 1-5 were transiently expressed in NIH 3T3 cells stably transformed with Ha-Ras(G12V) to assess the ability of each receptor to stimulate protein tyrosine phosphatase (PTPase) activity in vitro. Treatment of membranes from sstr2-, sstr3-, or sstr4-expressing cells with somatostatin-14 plus guanyl-5'-yl imidodiphosphate (GMPPNP) increased PTPase activity, and this stimulation was pertussis toxin-sensitive. Somatostatin alone, GMPPNP alone, or somatostatin plus GDP were ineffective under these conditions. sstr1 and sstr5 failed to increase PTPase activity although both receptors were expressed, as assessed by appearance of high-affinity binding sites for [125I Tyr11]somatostatin-14. Somatostatin plus GMPPNP stimulated PTPase activity in vitro when sstr2 was coexpressed with wild type PTP1B or a Cys to Ser (C/S), catalytically inactive PTP1B or with wild type SH2-domain containing PTPase SHP 2. However, coexpression with catalytically inactive C/S SHP-2 abrogated this response. Thus, three of the five cloned sstr's can couple to activate PTPase in this cellular background. Abrogation of the response by C/S SHP-2 strongly suggests, but does not prove, a role for SHP-2 in the mechanism. PMID- 9212055 TI - Novel isoforms of Mel1c melatonin receptors modulating intracellular cyclic guanosine 3',5'-monophosphate levels. AB - Two cDNAs encoding novel isoforms of Xenopus laevis melatonin receptors were cloned using PCR primers specific for the X. laevis-melanophore Mel1c melatonin receptor described in a recent publication. The novel isoforms were highly homologous to the described frog Mel1c cDNA, although the C-terminal tail of both was shorter by 65 amino acid residues. Nucleotide sequences of these novel isoforms, called Mel1c(alpha) and Mel1c(beta), differed from each other by only 35 nucleotides and six amino acid residues. Studies on several animals of various Xenopus species indicate that Mel1c(alpha) and Mel1c(beta) receptors may correspond to allelic variants of the same locus. Studies on cells transfected with both receptor cDNAs showed the expression of high-affinity 2 [125I]iodomelatonin binding sites. Agonist stimulation of Mel1c(alpha) receptor was associated with the inhibition of cAMP accumulation stimulated by forskolin (IC50 approximately 10(-10) M) in HeLa, Ltk-, and human embryonic kidney 293 (HEK 293) cells. Mel1c(beta) receptor modulated cAMP in HeLa and HEK 293 cells but not in Ltk- cells. Both receptors inhibited, in a dose-dependent manner, cGMP accumulation in all three cell lines incubated with a phosphodiesterase inhibitor. This effect was localized upstream of soluble guanylyl cyclase and was blocked by pertussis toxin treatment. However, IC50 values (approximately 10(-10) M for Mel1c(beta) and 10(-9) to 10(-7) M for Mel1c(alpha)) and maximal inhibition levels showed that Mel1c(alpha) receptors are much less efficiently coupled to the cGMP pathway. Coupling differences may be explained by the fact that five of the six amino acid substitutions between Mel1c(alpha) and Mel1c(beta) receptors are located within cytoplasmic regions potentially involved in signal transduction. The existence of coupling differences is in agreement with the observation that expression of both receptors is evolutionally conserved in native tissue. In conclusion, two novel, potentially allelic, isoforms of Xenopus Mel1c melatonin receptors display identical ligand-binding characteristics, but different potencies in modulating cAMP and cGMP levels through G(i)/G(o) dependent pathways. Furthermore, to our knowledge, this study provides the first data on the modulation of intracellular cGMP levels by cloned melatonin receptors. PMID- 9212056 TI - A GT box element is essential for basal and cyclic adenosine 3',5'-monophosphate regulation of the human surfactant protein A2 gene in alveolar type II cells: evidence for the binding of lung nuclear factors distinct from Sp1. AB - The gene encoding surfactant protein-A (SP-A) is developmentally regulated in type II cells of the fetal lung. In humans there are two SP-A genes, SP-A1 and SP A2. The SP-A2 gene is more highly regulated by cAMP and during fetal development than SP-A1. In earlier studies we determined that 296 bp of sequence flanking the 5'-end of the SP-A2 gene is sufficient to mediate high basal and cAMP-inducible reporter gene expression in primary cultures of transfected type II cells, suggesting that this region contains important cis-acting elements involved in tissue-specific and hormonal regulation of SP-A2 promoter activity. We also observed that mutagenesis of a cAMP response element (CRE)-like sequence at -242 bp (CRE(SP-A2)) greatly reduced basal and cAMP-stimulated expression in transfected type II cells. In the present study, we identified a GT box (GGGGTGGGG) at -61 bp of SP-A2 5'-flanking sequence that is highly conserved among the SP-A genes of different species. In type II cell transfection studies, we found that mutagenesis of the GT box of SP-A2 markedly reduced basal and abolished cAMP-induced reporter gene expression. Thus, CRE(SP-A2) and the GT box cooperatively interact to mediate basal and cAMP induction of SP-A2 promoter activity in type II cells. By electrophoretic mobility shift assays (EMSA), it was observed that nuclear proteins isolated from primary cultures of type II cells bound the GT box as five specific complexes. By contrast, nuclear proteins isolated from lung fibroblasts displayed notably reduced binding activity. Competition and supershift EMSA indicate that the ubiquitously expressed transcription factor Sp1, a GC box-binding protein of approximately 100 kDa, is a component of the complex of proteins that bind the GT box of SP-A2. The finding that only two of the five GT box-binding complexes were supershifted by incubation with Sp1 antibody suggests that a factor(s) in type II cell nuclear extracts that is distinct from Sp1 also interacts with the GT box. By UV cross linking and SDS-PAGE/EMSA analysis, we have identified a approximately 55-kDa GT box-binding factor in type II cell nuclear proteins that preferentially binds the GT box of SP-A2 over the consensus Sp1 GC box sequence. This 55-kDa factor was able to bind the GT box independently of Sp1. PMID- 9212057 TI - Internalization and homologous desensitization of the GLP-1 receptor depend on phosphorylation of the receptor carboxyl tail at the same three sites. AB - Homologous desensitization and internalization of the GLP-1 receptor correlate with phosphorylation of the receptor in a 33-amino acid segment of the cytoplasmic tail. Here, we identify the sites of phosphorylation as being three serine doublets located at positions 441/442, 444/445, and 451/452. The role of phosphorylation on homologous desensitization was assessed after stable expression in fibroblasts of the wild type or of mutant receptors in which phosphorylation sites were changed in various combinations to alanines. We showed that desensitization, as measured by a decrease in the maximal production of cAMP after a first exposure of the cells to GLP-1, was strictly dependent on phosphorylation. Furthermore, the number of phosphorylation sites correlated with the extent of desensitization with no, intermediate, or maximal desensitization observed in the presence of one, two, or three phosphorylation sites, respectively. Internalization of the receptor-ligand complex was assessed by measuring the rate of internalization of bound [125I]GLP-1 or the redistribution of the receptor to an endosomal compartment after agonist binding. Our data demonstrate that internalization was prevented in the absence of receptor phosphorylation and that intermediate rates of endocytosis were obtained with receptors containing one or two phosphorylation sites. Thus, homologous desensitization and internalization require phosphorylation of the receptor at the same three sites. However, the differential quantitative impairment of these two processes in the single and double mutants suggests different molecular mechanisms controlling desensitization and internalization. PMID- 9212058 TI - Ascorbic acid-dependent activation of the osteocalcin promoter in MC3T3-E1 preosteoblasts: requirement for collagen matrix synthesis and the presence of an intact OSE2 sequence. AB - Osteocalcin is a hormonally regulated calcium-binding protein made almost exclusively by osteoblasts. In normal cells, osteocalcin expression requires ascorbic acid (AA), an essential cofactor for osteoblast differentiation both in vivo and in vitro. To determine the mechanism of this regulation, subclones of MC3T3-E1 preosteoblasts were transiently transfected with 1.3 kb of the mouse osteocalcin gene 2 promoter driving expression of firefly luciferase. AA stimulated luciferase activity 20-fold after 4-5 days. This response was stereospecific to L-ascorbic acid and was only detected in MC3T3-E1 subclones showing strong AA induction of the endogenous osteocalcin gene. Similar results were also obtained in MC3T3-E1 cells stably transfected with the osteocalcin promoter. A specific inhibitor of collagen synthesis, 3,4-dehydroproline, blocked AA-dependent induction of promoter activity, indicating that regulation of the osteocalcin gene requires collagen matrix synthesis. Deletion analysis of the mOG2 promoter identified an essential region for AA responsiveness between -147 and -116 bp. This region contains a single copy of the previously described osteoblast-specific element, OSE2. Deletion and mutation of OSE2 in DNA transfection assays established the requirement for this element in the AA response. Furthermore, DNA-binding assays revealed that MC3T3-E1 cells contain OSF2, the nuclear factor binding to OSE2, and that binding of OSF2 to OSE2 is up regulated by AA treatment. Taken collectively, our results indicate that an intact OSE2 sequence is required for the induction of osteocalcin expression by AA. PMID- 9212059 TI - Hinge and amino-terminal sequences contribute to solution dimerization of human progesterone receptor. AB - We and others have shown previously that progesterone receptors (PR) form homodimers in solution in the absence of DNA and that dimers are the preferential form of receptor that binds with high affinity to target DNA. To determine the sequence regions involved in solution homodimerization, wild type PR and truncated PR proteins were expressed in an insect baculovirus system. The expression constructs included the ligand-binding domain [LBD, amino acids (aa) 688-933], the LBD plus hinge (hLBD, aa 634-933), the hLBD plus the DNA-binding domain (DhLBD, aa 538-933), and the full- length A and B isoforms of PR. PR-PR interactions were detected by three methods, coimmunoprecipitation of the PR fragments with full-length PR-A, pull-down of PR-polypeptides with polyhistidine tagged versions of the same polypeptides immobilized to metal affinity columns and cooperative ligand-binding assays (Hill coefficient, n(H) > 1 indicating PR PR interaction). By all three assays, the LBD alone was not sufficient to mediate protein-protein interaction. However, the LBD did exhibit other properties ascribed to this domain, including binding to steroids with a relatively good affinity and specificity, ligand-induced conformational changes at the carboxyl terminus tail and binding of heat shock protein 90 and its dissociation in response to hormone. Thus, failure of the expressed LBD to mediate dimerization does not appear to be due to an extensively misfolded or unstable polypeptide. The minimal carboxyl-terminal fragment capable of mediating PR-PR interaction was the hLBD construct. However, by immobilized metal affinity chromatography assay, self-association of PR-A was 3.5-fold more efficient than that of either the DhLBD or hLBD constructs. An expressed amino-terminal domain (aa 165-535) lacking the DNA-binding domain, hinge, and LBD was found to physically associate with PR A or with another amino-terminal fragment lacking the LBD, but retaining the DNA binding domain. These results provide evidence for direct amino-terminal interactions in the more efficient PR-PR interaction exhibited by wild-type PR-A, as compared with DhLBD and hLBD constructs. The overall results of this paper are consistent with the conclusion that the carboxyl-terminal LBD is not sufficient for mediating PR dimerization and that multiple regions, including the hinge and amino-terminal sequences, contribute either directly or indirectly to homodimerization of PR. PMID- 9212060 TI - Fibroblast growth factor receptor signaling activates the human interstitial collagenase promoter via the bipartite Ets-AP1 element. AB - Interstitial collagenases participate in the remodeling of skeletal matrix and are regulated by fibroblast growth factor (FGF). A 0.2-kb fragment of the proximal human interstitial collagenase [matrix metalloproteinase (MMP1)] promoter conveys 4- to 8-fold induction of a luciferase reporter in response to FGF2 in MC3T3-E1 osteoblasts. By 5'-deletion, this response maps to nucleotides 100 to -50 relative to the transcription initiation site. The 63- bp MMP1 promoter fragment -123 to -61 confers this FGF2 response on the rous sarcoma virus minimal promoter. Intact Ets and AP1 cognates in this element are both required for responsiveness. The AP1 site supports basal and FGF-inducible promoter activity. The intact Ets cognate represses basal transcriptional activity in both heterologous and native promoter contexts and is also required for FGF activation. FGF2 up-regulates a DNA-binding activity that recognizes the MMP1 AP1 cognate and contains immunoreactive Fra1 and c-Jun. Both constitutive and FGF-inducible DNA-binding activities are present in MC3T3-E1 cells that recognize the MMP1 Ets cognate; prototypic Ets transcriptional activators are not present in these complexes. Inhibitors of protein kinase C, phosphatidyl inositol 3-OH kinase, and calmodulin-dependent protein kinase do not attenuate MMP1 promoter activation. FGF2 activates ERK1/ERK2 signaling in osteoblasts; however, 25 microM MAPK-ERK kinase (MEK) inhibitor PD98059 (inhibits by > 85% the phosphorylation of ERK1/ERK2) has no effect on MMP1 promoter activation by FGF2. Ligand-activated and constitutively active FGF receptors initiate MMP1 induction. Dominant negative Ras abrogates MMP1 induction by constitutively active FGFR2 ROS, but dominant negative Rho and Rac do not inhibit induction. The mitogen activated protein kinase (MAPK) phosphatase MKP2 [inactivates extracellular regulated kinase (ERK) = Jun N-terminal kinase (JNK) > p38 MAPK] completely abrogates MMP1 activation, whereas PAC1 (inactivates ERK = p38 > JNK) attenuates but does not completely prevent induction. Thus, a Ras- and MKP2-regulated MAPK pathway, independent of ERK1/ERK2 MAPK activity, mediates FGF2 transcriptional activation of MMP1 in MC3T3-E1 osteoblasts, converging upon the bipartite Ets-AP1 element. The DNA-protein interactions and signal cascades mediating FGF induction of the MMP1 promoter are distinct from two other recently described FGF response elements: the MMP1 promoter (-123 to -61) represents a third FGF-activated transcriptional unit. PMID- 9212061 TI - Regulation of gonadotropin-releasing hormone (GnRH) gene expression by insulin like growth factor I in a cultured GnRH-expressing neuronal cell line. AB - A GnRH-expressing neuronal cell line (NLT) was used to determine whether insulin like growth factor I (IGF-I) regulates GnRH gene expression. A receptor-binding assay demonstrated the expression of IGF-I receptors on NLT cells. Activation of IGF-I receptors induced the Ras/Raf-1/mitogen-activated protein kinase pathway and increased c-fos expression. NLT cells treated with IGF-I underwent cell proliferation and exhibited a growth-independent increase in mouse GnRH mRNA expression. In cells transfected with DNA constructs containing the human GnRH promoter, which includes a consensus AP-1 binding site fused to the luciferase reporter gene, a significant increase in reporter activities was induced by IGF I, whereas mutation of this AP-1 site significantly reduced IGF-I-induced promoter activation. These results demonstrate that IGF-I serves as an important signal in the regulation of both human and rodent GnRH gene expression. PMID- 9212062 TI - Differential hormone-dependent transcriptional activation and -repression by naturally occurring human glucocorticoid receptor variants. AB - The molecular mechanisms underlying primary glucocorticoid resistance or hypersensitivity are not well understood. Using transfected COS-1 cells as a model system, we studied gene regulation by naturally occurring mutants of the glucocorticoid receptor (GR) with single-point mutations in the regions encoding the ligand-binding domain or the N-terminal domain reflecting different phenotypic expression. We analyzed the capacity of these GR variants to regulate transcription from different promoters, either by binding directly to positive or negative glucocorticoid-response elements on the DNA or by interfering with protein-protein interactions. Decreased dexamethasone (DEX) binding to GR variants carrying mutations in the ligand-binding domain correlated well with decreased capacity to activate transcription from the mouse mammary tumor virus (MMTV) promoter. One variant, D641V, which suboptimally activated MMTV promoter mediated transcription, repressed a PRL promoter element containing a negative glucocorticoid-response element with wild type activity. DEX-induced repression of transcription from elements of the intercellular adhesion molecule-1 promoter via nuclear factor-kappaB by the D641V variant was even more efficient compared with the wild type GR. We observed a general DEX-responsive AP-1-mediated transcriptional repression of the collagenase-1 promoter, even when receptor variants did not activate transcription from the MMTV promoter. Our findings indicate that different point mutations in the GR can affect separate pathways of gene regulation in a differential fashion, which can explain the various phenotypes observed. PMID- 9212063 TI - Structural organization of the human vitamin D receptor chromosomal gene and its promoter. AB - The vitamin D receptor (VDR) is known to mediate the pleiotropic biological actions of 1,25-dihydroxyvitamin D3 through its ability to modulate the expression of target genes. The regulation of this ligand-activated cellular transcription factor is reported to occur at both transcriptional and posttranslational levels. To begin to address the molecular basis by which the VDR gene is regulated transcriptionally, we report here an initial characterization of the human VDR gene and its promoter. We isolated several overlapping A-phage and cosmid clones that cover more than 100 kb of human DNA and contained the entire VDR gene. The gene is comprised of 11 exons that, together with intervening introns, span approximately 75 kb. The noncoding 5'-end of the gene includes exons 1A, 1B, and 1C. Eight additional exons (exons 2-9) encode the structural portion of the VDR gene product. While primer extension and S1 nuclease-mapping studies reveal several common transcriptional start sites, three unique mRNA species are produced as a result of the differential splicing of exons 1B and 1C. The DNA sequence lying upstream of exon 1A is GC rich and does not contain an apparent TATA box. Several potential binding sites for the transcription factor SP1 and other activators are evident. Fusion of DNA fragments containing putative promoter sequences upstream of the luciferase structural gene followed by transient transfection of these plasmids into several mammalian cell lines resulted in significant reporter activity. Due to the size and complexity of the 5'-end of the VDR gene, we examined the activity of a DNA fragment surrounding exon 1C. An intron fragment 3' of exon 1C conferred retinoic acid responsivity when fused to a reporter gene plasmid, suggesting a molecular mechanism for the previously observed ability of retinoic acid to induce the VDR. The recovery of the gene for the human VDR will enable further studies on the transcriptional regulation of this gene. PMID- 9212064 TI - JAK2 and STAT5, but not JAK1 and STAT1, are required for prolactin-induced beta lactoglobulin transcription. AB - Several different Janus kinases (JAKs) and signal transducers and activation of transcription (STATs) have been implicated in mediating the biological responses induced by PRL, based on their ligand-dependent tyrosine phosphorylation and activation. However, these criteria alone do not prove that a particular JAK or STAT is essential for signal transduction. We have used mutant cell lines defective in JAK1, JAK2, or STAT1 to examine their roles in PRL-dependent signaling. JAK2 is absolutely required for PRL-dependent phosphorylation of the receptor, activation of STATs, and induction of beta-lactoglobulin. Wild type, but not kinase-negative JAK2, restores all responses to PRL in JAK2-defective cells, suggesting that JAK2 function, not merely the protein, is required. In contrast, JAK1, which is phosphorylated in response to PRL, is not required for any of these functions. Although STAT1 homodimers do form in response to PRL, no defect in PRL-dependent signaling is apparent when STAT1 is missing, suggesting that STAT5, which is strongly activated in response to PRL, is primarily responsible for driving the expression of PRL-responsive genes. PMID- 9212065 TI - Synergistic effects of testosterone metabolites on the development of motoneuron morphology in a sexually dimorphic rat spinal nucleus. AB - The rat lumbar spinal cord contains the testosterone-dependent spinal nucleus of the bulbocavernosus (SNB), whose motoneurons innervate perineal muscles involved in copulatory reflexes. In normal males, SNB dendrites grow exuberantly through the first 4 weeks postnatally. This growth is steroid-dependent: dendrites fail to grow in males castrated at P7, but grow normally in castrates treated with testosterone (T). Treatment with either of the T metabolites, dihydrotestosterone or estrogen, supports dendritic growth in castrates, but not to the lengths characteristic of intact males or T-treated castrates. The present study tested the hypothesis that dihydrotestosterone and estrogen act together to support development of SNB dendrites. Male rat pups were castrated on P7 and treated daily with dihydrotestosterone propionate (DHT) (2 mg), estradiol benzoate (E) (100 microg), DHT (2 mg) combined with estradiol benzoate in either 5 microg (E5) or 100 microg (E100) doses, or vehicle alone. On P28, when SNB dendritic length is normally maximal, motoneurons were retrogradely labeled with cholera toxin-HRP (BHRP). Soma size and dendritic lengths of labeled motoneurons were assessed and compared to those of age-matched, intact male rats. Soma areas of DHT + E5 treated and DHT + E100-treated castrates did not differ from those of castrates treated with DHT alone, although somata of all three groups were significantly larger than those of normal males and E- or oil-treated castrates. Dendritic lengths in DHT + E5-treated castrates were significantly shorter than those of normal males, and did not differ from those of castrates receiving DHT or E alone, although all hormone-treated groups had dendritic lengths that were significantly longer than untreated castrates. However, treatment of castrates with DHT + E100 fully supported dendritic growth to levels characteristic of normal males. These results suggest that somal and dendritic growth may occur through separate developmental mechanisms, and that E and DHT act synergistically to support normal masculine SNB dendritic development. PMID- 9212066 TI - New electrical properties of neurons induced by a homeoprotein. AB - To explore possible neurogenic functions of the genes of the Hox/HOM complexes, we injected the mRNA from the leech homeobox genes Lox1 and Lox4 into adult neurons that normally do not express them. The ectopic expression of Lox1 induced a specific transformation in the electrical properties of certain identified neurons: action potential amplitude increased about threefold after the injections. This effect of Lox1 expression was restricted, among cell types examined, to the anterior pagoda neurons (APs) and the nut neurons. This effect was also restricted to Lox1 ectopic expression; the action potentials of APs and nut neurons were not enlarged when the mRNAs of either Lox4, another leech Hox/HOM gene, or beta-galactosidase were injected. Lox1 mRNA injection did not affect the resting potential, input resistance, or axonal morphology of the transformed APs, raising the possibility that it acts via the modification of voltage-dependent ion channels. Thus, a specific homeobox gene can transform key neuronal characteristics in a cell-specific manner. We may thus add electrophysiologic properties to other aspects of neuronal identity determined by homeobox gene expression. PMID- 9212067 TI - Analysis of spontaneous electrical activity in cerebellar Purkinje cells acutely isolated from postnatal rats. AB - Whole-cell patch recording techniques were used to analyze spontaneous electrical activity in cerebellar Purkinje cells acutely isolated from postnatal rats. Spontaneous activity was present in 65% of the cells examined, and it included simple and complex firing patterns which persisted under conditions that eliminated residual or reformed synaptic contacts. Under voltage clamp, both spontaneous and quiescent cells displayed similar voltage-dependent conductances. Inward current was carried by Na+ through tetrodotoxin (TTX)-sensitive channels and by Ca2+ through P-type and T-type Ca channels. P-type current was present in all cells examined. T-type current was found in <50%, and it did not correlate with spontaneous activity. We found no evidence of a transient (A-type) potassium current or hyperpolarization-activated cationic current in either spontaneous or quiescent cells. Spontaneous activity did correlate with a lower activation threshold of the Na current, resulting in substantial overlap of the activation and inactivation curves. TTX reduced the holding current of spontaneous cells clamped between -50 and -30 mV, consistent with the presence of a Na "window" current. We were unable, however, to measure a persistent component of the Na current using voltage steps, a result which may reflect the complex gating properties of Na channels. An Na window current could provide the driving force underlying spontaneous activity, as well as plateau potentials, in Purkinje cells. PMID- 9212068 TI - Neuronal cAMP-dependent protein kinase type II is concentrated in mushroom bodies of Drosophila melanogaster and the honeybee Apis mellifera. AB - In both Drosophila melanogaster and the honeybee Apis mellifera, cyclic adenosine monophosphate (cAMP)-dependent processes have been implicated in mechanisms of learning. This study characterizes the type II cAMP-dependent protein kinase (PKAII), the major target of cAMP in adult animals. In both species, PKAII is restricted to neuronal tissue, in which it accounts for more than 90% of total PKA activity. Although the intensity of PKAII immunoreactivity differs between distinct brain regions, labeling is detectable in all neuropiles and most somata. While the visual neuropiles, the antennal lobes, and structures of the central brain exhibit intermediate immunostaining, the mushroom bodies show high labeling and contain a three- to fourfold higher PKA activity compared to other neuropiles. Since the mushroom bodies are central sites of olfactory learning mediated via cAMP-dependent signaling, the modulatory functions of transmitters on PKA activity in Kenyon cells from the honeybee were tested. Agents which elevate cytoplasmic Ca2+ levels have no effects on PKA activity in cultured Kenyon cells. Dopamine, serotonin, and octopamine, however, cause an increase in PKA activity in Kenyon cells. The modulation of PKA activity by octopamine, the putative transmitter of the unconditioned stimulus in associative olfactory learning in the honeybee, together with the findings on the central role of the cAMP cascade in Drosophila mushroom bodies, suggests a major implication of PKAII mediated phosphorylation in learning and memory in both Drosophila and Apis. PMID- 9212069 TI - Vasotocinergic innervation of areas containing aromatase-immunoreactive cells in the quail forebrain. AB - In the male quail forebrain, aromatase-immunoreactive (ARO-ir) elements are clustered within the sexually dimorphic medial preoptic nucleus (POM), nucleus striae terminalis (nST), nucleus accumbens (nAc), and ventromedial and tuberal hypothalamus. These ARO-ir cells are sensitive to testosterone and its metabolites: Their number and size increase after exposure to these steroids. The POM and lateral septum are also characterized by a dense vasotocinergic innervation that is also sensitive to testosterone. We analyzed here the anatomical relationships between ARO-ir elements and VT-ir fibers in the quail prosencephalon. Sequential staining for vasotocin, aromatase, or vasotocin plus aromatase was performed on adjacent 30-microm-thick cryostat sections. High concentrations of thin VT-ir fibers were observed within the POM, nST, lateral septum, periventricular mesencephalic central gray, and ventromedial and tuberal hypothalamus. There was a close correspondence between the extension of the ARO ir cells and of VT-ir fibers. In double-labeled sections, all clusters of ARO-ir cells with the exception of those located in the nAc were embedded in a dense network of VT-ir fibers. Many of the VT-ir terminals appeared to end in the neuropile surrounding ARO-ir elements rather than directly on their cell bodies. This study supports the idea that the testosterone-dependent aromatase system is directly innervated by a testosterone-dependent peptidergic system. Aromatase containing cells could therefore be modulated by steroids both directly and indirectly through the vasotocin system. Alternatively, this neuroanatomical arrangement may mediate the control of vasotocin synthesis or release by steroids. Functional studies demonstrate that both aromatase and vasotocin affect reproductive behavior in quail, and the present data provide anatomical support for the integration of these effects. PMID- 9212070 TI - Sexually dimorphic neuron addition to an avian song-control region is not accounted for by sex differences in cell death. AB - Only male zebra finches sing, and several brain regions implicated in song behavior exhibit marked sex differences in neuron number. In one region, the high vocal center (HVC), this dimorphism develops because the incorporation of new neurons is greater in males than in females during the first several weeks after hatching. Although estrogen (E2) exposure stimulates neuron addition in females, it is not known where (E2) acts, or to what extent sexual differentiation influences the production, specification, or survival of HVC neurons. In the present study we first reassessed sex and (E2)-induced differences in cell degeneration within the HVC using the TUNEL technique to identify cells undergoing DNA fragmentation indicative of apoptosis. HVC neuron number, as well as the density and number of TUNEL-labeled and pyknotic cells within the HVC were measured in normal 20- and 30-day-old males and females, and in 30-day-old females implanted with E2 on posthatch day 18. Although HVC neuron number was greater in males than in females, and was masculinized in E2 females, no group differences were evident in the absolute number of dying cells. These results indicate that sex differences in cell survival within the HVC do not entirely account for sexually dimorphic neuron addition to this region. Rather, sexual differentiation acts on some HVC neurons before they complete their migration and/or early differentiation. Although the migratory route of HVC neurons is not known, a large number of E2 receptor-containing cells (ER cells) reside just ventromedial to the HVC and adjacent to the proliferative ventricular zone. Next, we investigated whether these ER cells contribute to early-arising sex differences in HVC neuron addition. By combining [3H]thymidine autoradiography with immunocytochemistry for ERs, we first established that ER-expressing cells are not generated during posthatch sexually dimorphic HVC neuron addition, and thus are not young HVC neurons that transiently express ERs during their migration. Furthermore, in 25-day-old birds we found no sex difference in the density of pyknotic cells among this group of ER cells, suggesting that these cells do not promote the differential survival of HVC neuronal precursors migrating through this region. Rather, ER cells or other cell populations may establish sex differences in HVC neuron number by creating dimorphisms in cellular specification. PMID- 9212071 TI - Is laminin-1 a guidance cue for cerebellar granule cell migration? AB - Laminin-1 is a glycoprotein found in the basement membrane of many tissues. In the cerebellum of rodents, it has also been localized along Bergmann glial fibers, where it is thought to be involved in promoting granule cell migration by enhancing adhesion and neurite outgrowth along these fibers. Recent reports, however, indicate that laminin-1 is not present on Bergmann fibers, but instead is associated with blood vessels and meninges. Furthermore, attempts to block granule cell migration using antibodies against laminin-1 have yielded conflicting results. In this report, we provide further evidence that laminin-1 is associated exclusively with blood vessels and meninges in the cerebellum of postnatal rats. In addition, we show that adhesion and neurite outgrowth of granule cells was impeded on laminin-coated surfaces. In fact, cerebellar cells dramatically and consistently avoided laminin-1 regions of patterned surfaces. Cells did adhere to laminin regions if it was coadsorbed with polylysine or tested in serum-containing medium. Avoidance of laminin-1 regions in culture was not, however, blocked by pretreatment with laminin-1 antibodies. By comparison, mouse neuroblastoma cells adhered preferentially to laminin-1 regions in serum free medium, a response which was blocked by laminin-1 antibodies. These results indicate that laminin-1 is not involved in granule cell migration along Bergmann glial fibers. Instead, they suggest that laminin-1 may function as a repulsive guidance cue preventing granule cells from following inappropriate pathways during development. PMID- 9212072 TI - Differential effects of depolarization on the growth of crayfish tonic and phasic motor axons in culture. AB - Previous studies have demonstrated neuron-specific differences in the inhibitory effects of depolarization upon neurite outgrowth. We examined whether there is a relationship between the normal impulse activity level of an axon and the effect of depolarization upon its growth. Inactive phasic motor axons and active tonic motor axons grow from crayfish abdominal nerve cord explants in culture. Depolarization of these axons with high K solutions produced greater inhibition of advancing growth cones from the phasic axons than from the tonic axons. During the period 20-40 min after the beginning of depolarization, tonic axon growth cones continued to advance, whereas phasic axon growth cones retracted. During chronic depolarization, all of the phasic axons retracted during the first day and approximately half of the phasic axons had degenerated after 4 days of depolarization. The majority of tonic axons continue to grow after 3 days of depolarization, and all of the tonic axon growth survived the 4 days of depolarization. The different responses of the growing phasic and tonic axons to depolarization appear to be Ca2+ dependent. The inhibitory effects of depolarization upon phasic axon growth were reduced by the Ca2+ channel blockers La3+ and Mg2+. Application of a Ca2+ ionophore, A23187, produces greater inhibition of phasic axon growth than tonic axon growth. This study demonstrates that depolarization produces greater inhibition of growth from inactive motor axons than from active motor axons. This is likely due to differences in Ca2+ regulation and/or sensitivity to intracellular Ca2+. PMID- 9212073 TI - Germany: new beginnings for biotechnology. PMID- 9212074 TI - Hepatic canalicular membrane 5: Expression and localization of the conjugate export pump encoded by the MRP2 (cMRP/cMOAT) gene in liver. AB - The liver converts endogenous and xenobiotic lipophilic compounds into anionic conjugates with glutathione, glucuronate, or sulfate. These conjugates are transported across the canalicular (apical) membrane into bile by a 190 kDa membrane glycoprotein that has been cloned recently. This apical conjugate transporting ATPase has been termed canalicular multidrug resistance protein (cMRP) because of the similarity in substrate specificity and sequence with the multidrug resistance protein (MRP1), canalicular multispecific organic anion transporter (cMOAT), or multidrug resistance protein 2 (MRP2). The amino acid sequence identity of human MRP2 and MRP1 is 49%. MRP2 is predominantly expressed in hepatocytes and localized to apical membrane domains. MRP2 is not expressed in the human Dubin-Johnson syndrome, which is therefore associated with an inherited deficiency in the secretion of amphiphilic anionic conjugates into the bile. The rat homolog Mrp2 is absent in two mutant strains of rats with different point mutations in the corresponding gene. These mutant rats are hyperbilirubinemic and deficient in the ATP-dependent transport of conjugates from hepatocytes into bile. Impairment of bile flow (cholestasis) can be associated with a down regulation of the expression of the conjugate export pump, and MRP2 contributes to bile flow as an important driving force. PMID- 9212075 TI - Sulfation and sulfotransferases 6: Biochemistry and molecular biology of plant sulfotransferases. AB - It is now well established that, in mammals, sulfate conjugation constitutes an important reaction in the transformation of xenobiotics and in the modulation of the biological activity of steroid hormones and neurotransmitter. The presence of a sulfate group on some molecules can also be a prerequisite for their biological function. For example, it is well known that the sulfate groups are directly involved in the molecular interaction between heparin and antithrombin III. In plants, sulfation also seems to play an important role in the intermolecular recognition and signaling processes, as indicated by the requirement of a sulfate moiety for the biological activity of gallic acid glucoside sulfate in the seismonastic and gravitropic movements of plants, and of Nod RM1 in the cortical cell division during early nodule initiation in Rhizobium meliloti-alfalfa interaction. In addition, recent studies indicate that flavonoid conjugates, including the sulfate esters, may play a role in the regulation of plant growth by strongly binding the naphthylphthalamic acid receptor, thus blocking the quercetin-stimulated accumulation of the auxin phytohormone. Although several sulfated metabolites are known to accumulate in a variety of plant species, the study of enzymes that catalyze the sulfation reaction in plants lagged considerably compared to those conducted with their mammalian homologs. This apparent lack of interest may have been because the function of plant-sulfated metabolites is difficult to predict, since their accumulation is often restricted to a limited number of species. Despite this limitation, several plant sulfotransferases (STs) have been characterized at the biochemical level, and the cDNA clones encoding six plant STs have been isolated. Based on sequence homology, the plant ST coding sequences are grouped under the SULT3 family, also known as the flavonol ST family. This review summarizes our current knowledge of the plant STs and focuses on the functional significance of the sulfate conjugation in plant growth, development, and adaptation to stress. PMID- 9212076 TI - Degradation of oxidized proteins in mammalian cells. AB - Protein oxidation in vivo is a natural consequence of aerobic life. Oxygen radicals and other activated oxygen species generated as by-products of cellular metabolism or from environmental sources cause modifications to the amino acids of proteins that generally result in loss of protein function/enzymatic activity. Oxidatively modified proteins can undergo direct chemical fragmentation or can form large aggregates due to covalent cross-linking reactions and increased surface hydrophobicity. Mammalian cells exhibit only limited direct repair mechanisms and most oxidized proteins undergo selective proteolysis. The proteasome appears to be largely responsible for the degradation of soluble intracellular proteins. In most cells, oxidized proteins are cleaved in an ATP and ubiquitin-independent pathway by the 20 S "core" proteasome. The proteasome complex recognizes hydrophobic amino acid residues, aromatic residues, and bulky aliphatic residues that are exposed during the oxidative rearrangement of secondary and tertiary protein structure: increased surface hydrophobicity is a feature common to all oxidized proteins so far tested. The recognition of such (normally shielded) hydrophobic residues is the suggested mechanism by which proteasome catalyzes the selective removal of oxidatively modified cell proteins. By minimizing protein aggregation and cross-linking and by removing potentially toxic protein fragments, proteasome plays a key role in the overall antioxidant defenses that minimize the ravages of aging and disease. PMID- 9212077 TI - New directions in breast cancer research. AB - Research in breast cancer extends in many directions, stimulated by concerns related to the high incidence of the disease and the relative unpredictability of its clinical course. Examples of work in several directions are presented here arranged by four levels of analysis. 1) Molecular, intracellular events (molecular genetics). Recent identification of genes that predispose to breast cancer, and the isolation of those genes and their protein products, permit investigations of the most critical issues: the roles of these genes in normal development and breast differentiation, and how their alteration permits or contributes to tumor initiation. Thus, we expect that understanding the functions of the genes involved in inherited susceptibility to breast cancer will also be informative for sporadic breast cancers. 2) Cellular biology (cellular models for preneoplastic disease). We examine models of breast cancer development and ask how they help to validate a morphologic sequence for human breast neoplasia and whether they permit investigation of how to modify disease progression. Two useful models, one in transgenic mice and the other using human breast stem cells capable of culture and xenograft growth, are now available. 3) Tissue and organ (the tumor and its local environment). We look at the relationship of the tumor cell population to its local environment (stroma, blood vessels, etc.). This leads naturally to questions of how neighboring tissues and cytokines may modify tumor growth. 4) The individual as an organism and member of a population (hormonal rise and chemoprevention). We address identification of the primarily hormonal risk factors and a possible related mode of cancer prevention. PMID- 9212078 TI - The role of glucose 6-phosphate in the control of glycogen synthase. AB - Elevated blood glucose concentrations result in increased intracellular levels of glucose 6-phosphate in liver, skeletal muscle, and adipose tissue. In liver, blood glucose concentrations are the main factor in control of the synthesis of glycogen; insulin has only a potentiating effect. In skeletal muscle and adipocytes, glucose alone has little effect on the activity of glycogen synthase, the limiting enzyme in glycogen synthesis. However, insulin released as a result of elevated blood glucose stimulates the translocation of specific glucose transporters to the cell membrane, increases the uptake of glucose, and causes the covalent, dephosphorylation-mediated activation of glycogen synthase. We present evidence that elevated intracellular contents of glucose 6-phosphate provoke the activation of glycogen synthase in liver, muscle, and adipose tissue. In addition, glucose 6-phosphate may inhibit the phosphorylation of glycogen synthase by cyclic AMP-stimulated protein kinase. We show that the stimulated glucose uptake and phosphorylation appear to play a major role in the control by insulin of the enzymes involved in glycogen synthesis. PMID- 9212079 TI - Cyclosporin A but not FK506 inhibits thyroid hormone-induced apoptosis in tadpole intestinal epithelium. AB - Amphibian metamorphosis and mammalian T cell development represent two of the best known systems where developmental programmed cell death through apoptosis takes place. Two immunosuppressants, cyclosporin A (CsA) and FK506, have been demonstrated to inhibit activation-induced cell death in immature T cells and T cell hybridomas. In this study, we have established an in vitro system in which isolated primary tadpole intestinal epithelial cells undergo typical apoptosis upon treatment with thyroid hormone (T3), the causative agent of metamorphosis. It is surprising that this T3-induced apoptosis was found to be inhibited only by CsA but not by FK506, whereas both immunosuppressants block activation-induced apoptosis in T cells. Since T3 exerts its effect primarily by regulating gene transcription through direct binding to nuclear thyroid hormone receptors, our results strongly suggest that except for their similarity in the T cell receptor mediated signal transduction process, CsA, but not FK506, also blocks another yet unidentified step during the induction of apoptosis. The identification of this novel function of CsA may provide an important clue toward the understanding of the mechanism of apoptosis and helps in designing better clinical applications of the immunosuppressants. PMID- 9212080 TI - Na+/Ca2+ exchanger modulates kainate-triggered Ca2+ signaling in Bergmann glial cells in situ. AB - The role of sodium-calcium exchanger in calcium homeostasis in Bergmann glial cells in situ was investigated by monitoring cytoplasmic calcium ([Ca2+]i) and sodium ([Na+]i) concentrations. The [Ca2+]i and [Na+]i transients were measured either separately by using fluorescent indicators fura-2 and SBFI, respectively, or simultaneously using the indicators fluo-3 and SBFI. Since the removal of extracellular Na+ induced a relatively small (approximately 50 nM) elevation of [Ca2+]i, the Na+/Ca2+ exchanger seems to play a minor role in regulation of resting [Ca2+]i. In contrast, kainate-triggered [Ca2+]i increase was significantly suppressed by lowering of the extracellular Na+ concentration ([Na+]o). In addition, manipulations with [Na+]o dramatically affected the recovery of the kainate-induced [Ca2+]i transients. Simultaneous recordings of [Ca2+]i and [Na+]i revealed that kainate-evoked [Ca2+]i transients were accompanied with an increase in [Na+]i. Moreover, kainate induced significantly larger [Ca2+]i and smaller [Na+]i transients under current-clamp conditions as compared to those recorded when the membrane voltage was clamped at -70 mV. The above results demonstrate that the Na(+)-Ca2+ exchanger is operative in Bergmann glial cells in situ and is able to modulate dynamically the amplitude and kinetics of [Ca2+]i signals associated with an activation of ionotropic glutamate receptors. PMID- 9212081 TI - Caloric restriction reduces fiber loss and mitochondrial abnormalities in aged rat muscle. AB - The influence of caloric restriction (CR) initiated at 17 months of age was investigated on selected age-associated measures in skeletal muscle. Tissue from young (3-4 months) ad libitum-fed, old (30-32 months) restricted (35% and 50% CR, designated CR35 and CR50, respectively), and old ad libitum-fed rats (29 months) was studied. CR preserved fiber number and fiber type composition in the vastus lateralis muscle of the CR50 rats. In the old rats from all groups, individual fibers were found with either no detectable cytochrome c oxidase activity (COX-), hyperreactivity for succinate dehydrogenase activity (SDH++; also known as ragged red fibers [RRF]), or both COX- and SDH++. Muscle from the CR50 rats contained significantly fewer COX- and SDH++ fibers than did the muscle from CR35 rats. CR50 rats also had significantly lower numbers of mtDNA deletion products in two (adductor longus and soleus) of the four muscles examined compared to CR35 rats. These data indicate that CR begun in late middle age can retard age-associated fiber loss and fiber type changes, as well as increases in the number of skeletal muscle fibers showing mitochondrial enzyme abnormalities. CR also decreased the accumulation of mtDNA deletions. PMID- 9212082 TI - Chimeric strategies for the rational design of bioactive analogs of small peptide hormones. AB - The aim of this study was to evaluate a variety of synthetic strategies pertinent to the development of chimeric analogs of the structurally divergent nonapeptide hormones arginine vasopressin (AVP) and bradykinin (BK). Single-chain peptides combining AVP and BK directly, AVP(1-9)-BK(1-9) or via a flexible aminohexanoic acid (epsilonAhx) linker, AVP(1-9)-epsilonAhx-BK(1-9), bind with relatively high affinity to the bovine kidney medulla B2a bradykinin receptor (B2a BKR). Significantly, amino-terminal extended chimeric analogs of BK, including AVP(1-9) BK(1-9) and galanin(1-13)-BK(1-9), are functional B2 BKR agonists. These findings illustrate that chimeric peptides can activate G-protein-coupled receptors (GPCRs) in a manner analogous to that of endogenous monomeric agonists. Further development, combining the sequences of receptor subtype-selective antagonists, produced high-affinity chimeric antagonists of the V1a vasopressin receptor (V1a VPR) and the B2a BKR. We also determined the pharmacological characteristics of high-affinity chimeric hormone analogs derivatized with the membrane targeting function of mastoparan. Homodimers of an amino-terminal extended BK analog and a V1a-selective antagonist represent the first examples of new classes of B2 BKR and V1a VPR antagonists, respectively. These findings are discussed in relation to the GPCR binding site for small peptides and the development of novel biological probes and therapeutic agents. PMID- 9212083 TI - p53 Deficiency in liver reduces local control of survival and proliferation, but does not affect apoptosis after DNA damage. AB - Despite good evidence for p53 dysfunction in human hepatocellular carcinomas, little is known of the significance of p53 to normal hepatocytes and whether p53 dysfunction is relevant to early hepatocarcinogenesis. We have therefore examined the consequences of targeted p53 deficiency in hepatocytes for regulation of apoptosis, proliferation, and ploidy. p53 deficiency was silent in normal liver and did not affect progression from diploidy to polyploidy in the aging liver. However, in primary culture the absence of p53 resulted in increased hepatocyte proliferation indices and decreased sensitivity to proliferation inhibition by TGFbeta. Moreover, p53-deficient cells continued to survive and proliferate under conditions of minimal trophic support that led to growth arrest and apoptosis of wild-type cells. In vivo, p53-deficient mice had enhanced proliferative responses to both xenobiotic hepatomitogen and CCl4-induced liver necrosis, although lack of persistent proliferation showed that other control mechanisms are important. There was no simple relationship between p53 and apoptosis after DNA damage because UV irradiation led to p53-independent apoptosis, even though p53 was stabilized. However, p53 did couple DNA damage to growth arrest, and abnormal mitoses after gamma-irradiation of regenerating p53 null livers demonstrated circumstances where loss of G1 and G2 checkpoints may generate abnormal ploidy. Thus p53 becomes important when hepatocytes are released from G0 and stressed, sensitizing them to mitogen and cytokine regulators of cell cycle progression and apoptosis. Hence p53 deficiency is likely to be significant in an environment of persistent regenerative stimuli and unfavorable trophic support or in the presence of other enabling genetic lesions. This model is relevant to human hepatocarcinogenesis, which almost always occurs against a background of chronic hepatocellular destruction in hepatitis and cirrhosis. In that context, by reducing the need for cytokine support and disabling DNA damage-induced growth arrest, p53 deficiency should facilitate the expansion of preneoplastic clones in chronic liver disease. PMID- 9212084 TI - A novel explanation for fluctuations of ion current through narrow pores. AB - Fluctuation of ion current, between a high conductance and a low conductance state, through biological ion channels and pores is assumed to arise from conformational changes between an "open" and a "closed" configuration. Here we offer an additional mechanism that arises from changes in ionization of fixed charges within, or at the mouth of, a channel or pore. Our hypothesis, which is based on measurements of ion selectivity alongside ion current, applies to pores through some synthetic membranes and through channels-such as those created by certain toxins-that remain (at least partially) open in the low conductance state. It may also explain the phenomena of "open channel noise" and "substate behavior" that characterize several endogenous ion channels and should be considered when modeling the behavior of such channels. PMID- 9212085 TI - Oligonucleotides: extrapolating from in vitro to in vivo. PMID- 9212086 TI - Graft persistence in animal models of psoriasis. PMID- 9212088 TI - Who is really behind Clinton's AIDS vaccine crusade? PMID- 9212087 TI - HIV antivirals and immune recovery. PMID- 9212089 TI - The American Cancer Society versus tobacco. The first 40 years or the last? PMID- 9212090 TI - What went right: why is HIV a treatable infection? PMID- 9212091 TI - Molecular time machines. The first gene involved in mammalian circadian timekeeping has been identified. PMID- 9212092 TI - Gene therapy for lysosomal storage disease: a no-brainer? Transplants of fibroblasts secreting high levels of beta-glucuronidase decrease lesions in the brains of mice with Sly syndrome, a lysosomal storage disease. PMID- 9212093 TI - Double indemnity: p53, BRCA and cancer. p53 mutation partially rescues developmental arrest in Brca1 and Brca2 null mice, suggesting a role for familial breast cancer genes in DNA damage repair. PMID- 9212095 TI - Transglutaminase, gluten and celiac disease: food for thought. Transglutaminase is identified as the autoantigen of celiac disease. PMID- 9212094 TI - Serpents on the road to dementia and death. Accumulating evidence from several studies points to the normal function of presenilin 1 and suggests how the mutant protein contributes to deposition of amyloid plaques in Alzheimer's disease. PMID- 9212096 TI - FasL--too much of a good thing? Transplanted grafts of pancreatic islet cells engineered to express Fas ligand are destroyed not protected by the immune system. PMID- 9212097 TI - Anticonvulsant action of neuropeptide Y. Neuropeptide Y may act as an endogenous anticonvulsant through Y5 receptors suggesting a new target for antiepileptic drugs. PMID- 9212098 TI - Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. AB - On the subject of acute myeloid leukemia (AML), there is little consensus about the target cell within the hematopoietic stem cell hierarchy that is susceptible to leukemic transformation, or about the mechanism that underlies the phenotypic, genotypic and clinical heterogeneity. Here we demonstrate that the cell capable of initiating human AML in non-obese diabetic mice with severe combined immunodeficiency disease (NOD/SCID mice) - termed the SCID leukemia-initiating cell, or SL-IC - possesses the differentiative and proliferative capacities and the potential for self-renewal expected of a leukemic stem cell. The SL-ICs from all subtypes of AML analyzed, regardless of the heterogeneity in maturation characteristics of the leukemic blasts, were exclusively CD34++ CD38-, similar to the cell-surface phenotype of normal SCID-repopulating cells, suggesting that normal primitive cells, rather than committed progenitor cells, are the target for leukemic transformation. The SL-ICs were able to differentiate in vivo into leukemic blasts, indicating that the leukemic clone is organized as a hierarchy. PMID- 9212100 TI - High plasma HDL concentrations associated with enhanced atherosclerosis in transgenic mice overexpressing lecithin-cholesteryl acyltransferase. AB - A subset of patients with high plasma HDL concentrations have enhanced rather than reduced atherosclerosis. We have developed a new transgenic mouse model overexpressing human lecithin-cholesteryl acyltransferase (LCAT) that has elevated HDL and increased diet-induced atherosclerosis. LCAT transgenic mouse HDLs are abnormal in both composition and function. Liver uptake of [3H]cholesteryl ether incorporated in transgenic mouse HDL was reduced by 41% compared with control HDL, indicating ineffective transport of HDL-cholesterol to the liver and impaired reverse cholesterol transport. Analysis of this LCAT transgenic mouse model provides in vivo evidence for dysfunctional HDL as a potential mechanism leading to increased atherosclerosis in the presence of high plasma HDL levels. PMID- 9212099 TI - Fas ligand expression in islets of Langerhans does not confer immune privilege and instead targets them for rapid destruction. AB - Fas ligand is believed to mediate immune privilege in a variety of tissues, including the eye, testis, and a subset of tumors. We tested whether expression of Fas ligand on pancreatic islets either following adenoviral or germline gene transfer could confer immune privilege after transplantation. Islets were infected with an adenoviral vector containing the murine Fas ligand cDNA (AdFasL), and were transplanted into allogenic diabetic hosts. Paradoxically, AdFasL-infected islets underwent accelerated neutrophilic rejection. The rejection was T cell and B cell independent and required Fas protein expression by host cells, but not on islets. Similarly, transgenic mice expressing Fas ligand in pancreatic beta cells developed massive neutrophilic infiltrates and diabetes at a young age. Thus, Fas ligand expression on pancreatic islets results in neutrophilic infiltration and islet destruction. These results have important implications for the development of Fas ligand-based immunotherapies. PMID- 9212101 TI - Heritable disorder resembling neuronal storage disease in mice expressing prion protein with deletion of an alpha-helix. AB - Mice were constructed carrying prion protein (PrP) transgenes with individual regions of putative secondary structure deleted. Transgenic mice with amino terminal regions deleted remained healthy at >400 days of age, whereas those with either of carboxy-terminal alpha-helices deleted spontaneously developed fatal CNS illnesses similar to neuronal storage diseases. Deletion of either C-terminal helix resulted in PrP accumulation within cytoplasmic inclusions in enlarged neurons. Deletion of the penultimate C-terminal helix resulted in proliferation of rough endoplasmic reticulum. Mice with the C-terminal helix deleted were affected with nerve cell loss in the hippocampus and proliferation of smooth endoplasmic reticulum. Whether children with the human counterpart of this malady will be found remains to be determined. PMID- 9212102 TI - Hyperaccumulation of FAD-linked presenilin 1 variants in vivo. AB - Mutations in the presenilin 1 (PS1) and presenilin 2 (PS2) genes can cause Alzheimer's disease in affected members of the majority of early-onset familial Alzheimer's disease (FAD) pedigrees. PS1 encodes an ubiquitously expressed, eight transmembrane protein. PS1 is endoproteolytically processed to an amino-terminal derivative (approximately 27-28 kDa) and a carboxy-terminal derivative (approximately 17-18 kDa). These polypeptides accumulate to saturable levels in the brains of transgenic mice, independent of the expression of PS1 holoprotein. We now document that, in the brains of transgenic mice, the absolute amounts of accumulated N- and C-terminal derivatives generated from the FAD-linked PS1 variants in which Glu replaces Ala at codon 246 (A246E) or Leu replaces Met at codon 146 (M146L) accumulate to a significantly higher degree (approximately 40 50%) than the fragments derived from wild-type PS1. Moreover, the FAD-linked deltaE9 PS1 variant, a polypeptide that is not subject to endoproteolytic cleavage in vivo, also accumulates in greater amounts than the fragments generated from wild-type human PS1. Thus, the metabolism of PS1 variants linked to FAD is fundamentally different from that of wild-type PS1 in vivo. PMID- 9212103 TI - Powerful inhibition of kainic acid seizures by neuropeptide Y via Y5-like receptors. AB - Neuropeptide Y (NPY) is widely distributed in interneurons of the central nervous system (CNS), including the hippocampus and cerebral cortex, in concentrations exceeding those of any other known neuropeptides. Sequence data comparing different species show that NPY is highly conserved. This suggests a critical role in regulation of regional neuronal excitability. Kainic acid, a glutamate agonist at kainic acid receptors, causes severe limbic motor seizures culminating in status epilepticus. We here report that NPY administered into the lateral ventricle is a powerful inhibitor of motor as well as electroencephalographic (EEG) seizures induced by kainic acid. This effect was mediated via receptors with a pharmacological profile similar to the recently cloned rat Y5 receptor. The present study is the first to demonstrate that NPY possesses anticonvulsant activity. This is consistent with the concept that NPY is an endogenous anticonvulsant and suggests that agonists acting at Y5-like receptors may constitute a novel group of drugs in antiepileptic therapy. PMID- 9212104 TI - Adenoviral gene transfer of ciliary neurotrophic factor and brain-derived neurotrophic factor leads to long-term survival of axotomized motor neurons. AB - The neurotrophic factors ciliary neurotrophic factor and brain-derived neurotrophic factor can prevent motor neuron cell death during development and after nerve lesion in neonatal rodents. However, local and systemic application of these factors to newborn rats with damaged motor nerves rescues motor neurons only transiently during the first two weeks after axotomy. In order to test the effect of continuous delivery of these factors, the effect of localized injection of CNTF- or BDNF-transducing recombinant adenoviruses into the lesioned nerves was investigated. Under such conditions, survival of axotomized motor neurons is maintained for at least 5 weeks. This way of delivery corresponds to the physiological situation in adult rodents, under which endogenous CNTF is present in the cytosol of Schwann cells and BDNF expression is upregulated after nerve lesion, making these factors available to the damaged motor neurons. Recent results show that overexpression of muscle-derived neurotrophin-3 prevents degeneration of axons and motor endplates, but has only little effect on the number of motor neuron cell bodies in a murine animal model of motor neuron disease. Therefore, techniques suitable for tonic exposure to both nerve- and muscle-derived neurotrophic factors may have implications for the design of future therapeutic strategies against human motor neuron disease. PMID- 9212105 TI - Decreased lysosomal storage in the adult MPS VII mouse brain in the vicinity of grafts of retroviral vector-corrected fibroblasts secreting high levels of beta glucuronidase. AB - A deficiency of beta-glucuronidase (GUSB) causes the multisystem progressive degenerative syndrome, mucopolysaccharidosis (MPS) type VII (Sly disease), which includes mental retardation. Animal homologues of MPS VII (ref. 3, 4) are models for testing somatic gene transfer approaches to treat the central nervous system in this and other lysosomal storage disorders. Previous attempts to correct murine MPS VII by gene therapy have successfully treated lesions in some organs but not in the brain. Other experimental modalities have forestalled some disease progression in the brain, but only if done at birth, before the onset of severe lesions, when the animals are phenotypically normal. We tested whether therapeutic amounts of GUSB could be delivered to the diseased adult brain by transplanting cells engineered to super-secrete the normal enzyme for export to surrounding neural tissues. Lysosomal distention was cleared from neurons and glial cells in the vicinity of the grafts, showing that the secreted enzyme could reach the diseased cells and reverse lesions in the severely diseased brain. The ability to correct established lesions will be important for the treatment of many lysosomal storage diseases affecting the brain, because most patients are not diagnosed until lesions are advanced enough to affect phenotype or developmental milestones in early childhood, and some forms of the diseases do not become apparent until later in life. PMID- 9212107 TI - Mapping pathophysiological features of breast tumors by MRI at high spatial resolution. AB - Magnetic resonance imaging (MRI) is a noninvasive method that reveals anatomical details in vivo and detects lesions for diagnosis. Although standard breast MRI cannot clearly delineate breast cancer, contrast-enhanced MRI enables the detection of breast masses with high sensitivity. Dynamic studies demonstrated that malignant lesions were characterized by a faster signal enhancement rate than benign ones. Dynamic MRI of human breast cancer in mice revealed high heterogeneity in the distribution of contrast-enhanced curves and derived pathophysiological features, indicating the importance of high spatial resolution. With clinical MRI, it is difficult to achieve simultaneously high spatial and temporal resolution. In previous dynamic studies, the emphasis was on high temporal resolution and mainly empiric analyses. We describe here a new model-based method that optimizes spatial resolution by using only three time points, and yet characterizes tumor heterogeneity in terms of microvascular permeability and extracellular fraction. Mapping these pathophysiological features may aid diagnosis and prognosis assessment, while the high spatial resolution may improve the capacity to detect smaller lesions. The method was tested in human breast tumors implanted in mice and in a limited number of benign and malignant breast lesions of patients. PMID- 9212106 TI - Activation of cAMP-PKA signaling in vivo inhibits smooth muscle cell proliferation induced by vascular injury. AB - Injury of the arterial wall induces the formation of the neointima. This structure is generated by the growth of mitogenically activated smooth muscle cells of the arterial wall. The molecular mechanism underlying the formation of the neointima involves deregulated cell growth, primarily triggered by the injury of the arterial wall. The activated gene products transmitting the injury-induced mitogenic stimuli have been identified and inhibited by several means: transdominant negative expression vectors, antisense oligodeoxynucleotides, adenovirus-mediated gene transfer, antibodies and inactivating drugs. Results of our study show that local administration of 3',5'-cyclic AMP and phosphodiesterase-inhibitor drugs (aminophylline and amrinone) to rats markedly inhibits neointima formation after balloon injury in vivo and in smooth muscle cells in vitro. The growth inhibitory effect of aminophylline was completely reversed by the inhibition of cAMP-dependent protein kinase A (PKA). These findings indicate an alternative approach to the treatment of diseases associated with injury-induced cell growth of the arterial wall, as stimulation of cAMP signaling is pharmacologically feasible in the clinical setting. PMID- 9212108 TI - Induction of high levels of allogeneic hematopoietic reconstitution and donor specific tolerance without myelosuppressive conditioning. AB - Donor-specific tolerance induced by bone marrow transplantation (BMT) would allow organ allografting without chronic immunosuppressive therapy. However, the toxicity of conditioning regimens used to achieve marrow engraftment has precluded the clinical use of BMT for tolerance induction. We have developed a BMT strategy that achieves alloengraftment without toxic or myelosuppressive host conditioning. B6 mice received depleting anti-CD4 and anti-CD8 monoclonal antibodies, local thymic irradiation, and a high-dose (174 x 10(6)) of major histocompatibility (MHC)-mismatched B10.A bone marrow cells (BMCs) divided over days 0 through 4. High levels of donor cells were observed among white blood cells (WBCs) of all lineages. Permanent, multilineage mixed chimerism; donor specific skin-graft tolerance; and in vitro tolerance were observed in most animals. Large numbers of donor class II(high) cells were detected in thymuses of long-term chimeras, and their presence was associated with intrathymic deletion of donor-reactive host thymocytes. The treatment was not associated with significant myelosuppression, toxicity, or graft-versus-host disease (GVHD). Thus, high levels of allogeneic stem-cell engraftment can be achieved without myelosuppressive host conditioning. As stem-cell mobilization and in vitro culture techniques have increased the feasibility of administering high doses of hematopoietic cells to humans, this approach brings hematopoietic cell transplantation closer to clinical use for the induction of central deletional T cell tolerance. PMID- 9212109 TI - Induction of basal cell carcinoma features in transgenic human skin expressing Sonic Hedgehog. AB - Hedgehog (HH) signaling proteins mediate inductive events during animal development. Mutation of the only known HH receptor gene, Patched (PTC), has recently been implicated in inherited and sporadic forms of the most common human cancer, basal cell carcinoma (BCC). In Drosophila, HH acts by inactivating PTC function, raising the possibility that overexpression of Sonic Hedgehog (SHH) in human epidermis might have a tumorigenic effect equivalent to loss of PTC function. We used retroviral transduction of normal human keratinocytes to constitutively express SHH. SHH-expressing cells demonstrated increased expression of both the known HH target, BMP-2B, as well as bcl-2, a protein prominently expressed by keratinocytes in BCCs. These keratinocytes were then used to regenerate human skin transgenic for long terminal repeat-driven SHH (LTR SHH) on immune-deficient mice. LTR-SHH human skin consistently displays the abnormal specific histologic features seen in BCCs, including downgrowth of epithelial buds into the dermis, basal cell palisading and separation of epidermis from the underlying dermis. In addition, LTR-SHH skin displays the gene expression abnormalities previously described for human BCCs, including decreased BP180/BPAG2 and laminin 5 adhesion proteins and expression of basal epidermal keratins. These data indicate that expression of SHH in human skin recapitulates features of human BCC in vivo, suggest that activation of this conserved signaling pathway contributes to the development of epithelial neoplasia and describe a new transgenic human tissue model of neoplasia. PMID- 9212110 TI - Transgenic plants expressing autoantigens fed to mice to induce oral immune tolerance. AB - Oral administration of protein can induce antigen-specific immune hyporesponsiveness. However, the utility of oral tolerance to autoantigens in the treatment of autoimmune diseases may be limited when candidate autoantigens cannot be produced by conventional systems in quantities sufficient for clinical studies. Plants may be ideally suited for this purpose, as they can synthesize, glycosylate and assemble mammalian proteins to provide huge quantities of relatively low cost soluble proteins. Furthermore, edible transgenic plants could provide a simple and direct method of autoantigen delivery for oral tolerance. Therefore, the aim of this study was to determine whether a transgenic plant expression system was capable of synthesizing the diabetes-associated autoantigen, glutamic acid decarboxylase (GAD) in an immunogenic form and whether the oral administration of an autoantigen expressed by a plant could directly induce protective immune responses in a mouse model of diabetes. We show that a GAD-expressing transgenic plant, given as a dietary supplement, inhibits the development of diabetes in the non-obese diabetic (NOD) mouse. PMID- 9212111 TI - Identification of tissue transglutaminase as the autoantigen of celiac disease. AB - Celiac disease is characterized by small intestinal damage with loss of absorptive villi and hyperplasia of the crypts, typically leading to malabsorption. In addition to nutrient deficiencies, prolonged celiac disease is associated with an increased risk for malignancy, especially intestinal T-cell lymphoma. Celiac disease is precipitated by ingestion of the protein gliadin, a component of wheat gluten, and usually resolves on its withdrawal. Gliadin initiates mucosal damage which involves an immunological process in individuals with a genetic predisposition. However, the mechanism responsible for the small intestinal damage characteristic of celiac disease is still under debate. Small intestinal biopsy with the demonstration of a flat mucosa which is reversed on a gluten-free diet is considered the main approach for diagnosis of classical celiac disease. In addition, IgA antibodies against gliadin and endomysium, a structure of the smooth muscle connective tissue, are valuable tools for the detection of patients with celiac disease and for therapy control. Incidence rates of childhood celiac disease range from 1:300 in Western Ireland to 1:4700 in other European countries, and subclinical cases detected by serological screening revealed prevalences of 3.3 and 4 per 1000 in Italy and the USA, respectively. IgA antibodies to endomysium are particularly specific indicators of celiac disease, suggesting that this structure contains one or more target autoantigens that play a role in the pathogenesis of the disease. However, the identification of the endomysial autoantigen(s) has remained elusive. We identified tissue transglutaminase as the unknown endomysial autoantigen. Interestingly, gliadin is a preferred substrate for this enzyme, giving rise to novel antigenic epitopes. PMID- 9212112 TI - Disruption of monolayer integrity enables activation of a cystic fibrosis "bypass" channel in human airway epithelia. AB - Cystic fibrosis (CF) is a genetic disease characterized by marked reduction in Cl conductance across many epithelia. Two kinds of Cl- channels have been associated with CF. One channel, termed the cystic fibrosis transmembrane conductance regulator (CFTR), is directly coded by the CF gene. The other channel is an outwardly rectifying depolarization induced Cl- channel (ORDIC) that is distinguished from other outwardly rectifying chloride channels (ORCCs) because its activity is induced most reliably by patch excision and depolarization. An issue in current CF research is whether ORDIC channels are indirectly activated by CFTR to contribute a significant portion of apical membrane Cl- conductance in airway cells. We now show that ORDIC channels are readily activated in patches excised and depolarized from isolated cells, but are rarer or refractory to activation in patches from the apical membranes of confluent human airway epithelia. These findings have important implications for proposed therapies that would bypass the CFTR conductance by activating ORDIC channels. PMID- 9212113 TI - Sensitivity and reproducibility in adenoviral infectious titer determination. PMID- 9212115 TI - Lumbar muscle rhabdomyolysis after abdominal aortic surgery. AB - Lumbar muscle rhabdomyolysis has been very rarely reported after surgery. The aim of this study was to determine its incidence and main characteristics in a large population undergoing abdominal aortic surgery. Over a 21-mo period, 224 consecutive patients, 209 male and 15 female, mean age 65 +/- 10 yr, underwent abdominal aortic surgery (aortic aneurysm in 142 patients and occlusive aortic degenerative disease in 82 patients). Surgical incision was a midline incision with exaggerated hyperlordosis in 173 patients and a flank incision with a retroperitoneal approach in 51 patients. Postoperative rhabdomyolysis was diagnosed in 20 patients. In these patients, 9 (4%) experienced severe low back pain, and lumbar muscle rhabdomyolysis was confirmed by tomodensitometry (n = 6) or muscle biopsy (n = 3). The remaining 11 patients had lower limb muscle rhabdomyolysis. Rhabdomyolysis occurred after surgery of longer duration, which involved more frequent visceral artery reimplantation, with longer duration of aortic clamping and greater intraoperative bleeding. Lumbar rhabdomyolysis occurred in younger patients who were more frequently obese. On first postoperative day, the mean creatine kinase (CK) value was greater in lumbar rhabdomyolysis than in lower limb rhabdomyolysis (17,082 +/- 15,003 vs 3,313 +/- 3,120 IU/L, P < 0.05). Acute renal failure and postoperative death did not occur in patients with lumbar muscle rhabdomyolysis. Lumbar rhabdomyolysis was not a rare event after abdominal aortic surgery (4%). This syndrome was characterized by postoperative low back pain of unusual severity, which required analgesic therapy, and induced a very high increase in CK with typical findings at tomodensitometry or muscle biopsy but was not associated with postoperative renal failure. PMID- 9212114 TI - Predictive factors for usefulness of fiberoptic pulmonary artery catheter for continuous oxygen saturation in mixed venous blood monitoring in cardiac surgery. AB - The main goal of this prospective study was to identify among cardiac surgery patients, usually monitored through a standard pulmonary artery catheter (PAC), those in whom a fiberoptic catheter oximeter to measure oxygen saturation in mixed venous blood (SVO2 PAC) would be most useful. Data from 286 patients who underwent coronary artery bypass graft (50%) or valvular surgery were recorded, including ASA physical status, New York Heart Association (NYHA) classification, and Parsonnet score (PS). Hemodynamic events and SVO2 changes were collected intra- and postoperatively until weaning from mechanical ventilation. The anesthesiologist in charge graded the usefulness of SVO2 PAC, and another anesthesiologist carried out a blindly controlled overall evaluation. Usefulness was defined as the presence of a change in therapeutic maneuver triggered solely by continuous SVO2 data that would not have occurred based on other routine parameters. SVO2 was also considered useful if earlier recognition of significant adverse events occurred. SVO2 PAC was useful in 57% of the patients. Independent predictive factors (multivariate analysis) for the perioperative usefulness of SVO2 in the whole population consisted of ASA class > or = 4 (P < 10(-5); relative risk [RR] 1.78, 1.51-2.07), mitral surgery (P < 10(-4); RR 1.72, 1.4 2.02), and NYHA score > or = 3 (P < 0.01; RR 1.66, 1.35-2.05). Independent predictive factors for the perioperative usefulness of SVO2 in the coronary artery bypass graft population were NYHA score > or = 3 (P < 10(-5); RR 1.90, 1.42-2.55) and ASA class > or = 4 (P < 0.01; RR 1.99, 1.51-2.63). The presence of three stenosed coronary arteries showed borderline significance (P < 0.06). Independent predictive factors for perioperative usefulness of SVO2 in the valvular population were mitral pathology (P < 10(-5)) and ASA class > or = 4 (P < 0.01). The receiver operator characteristic curve assessed the predictivity of the PS. SVO2 PAC was more useful in the group of patients with the greatest severity of illness (PS in useful group 17.0 +/- 10.3; in nonuseful group 8.7 +/- 6.6; P < 10(-4)). Intensive care unit duration and hospital stay in the useful group was prolonged compared with the nonuseful group. Similarly, morbidity was frequent in the useful group, although it was not always significantly different from the nonuseful group according to the type of complications. Mortality was comparable in the groups despite their different degree of illness and was reduced when taking into account the predictive and observed mortality provided by the PS. This study defined independent preoperative factors associated with SVO2 PAC monitoring and proposed a cutoff point above which SVO2 may be useful. PMID- 9212116 TI - The effect of milrinone on hemodynamics and left ventricular function after emergence from cardiopulmonary bypass. AB - Although milrinone effectively increases cardiac function, few studies have specifically evaluated its efficacy during cardiac surgery. We investigated the effects of milrinone on hemodynamics and left ventricular function in cardiac surgical patients who were already treated with catecholamines. Thirty-seven patients undergoing cardiac surgery were studied. Immediately after emergence from cardiopulmonary bypass (CPB), patients were randomly assigned to a control group (n = 10) or to one of these milrinone groups: milrinone 50 microg/kg intravenously (n = 8), 50 microg/kg + 0.5 microg x kg(-1) x min(-1) (n = 10), or 75 microg/kg + 0.75 microg x kg(-1) x min(-1) (n = 9). Hemodynamics and transesophageal echocardiogram were recorded while constant filling pressures were maintained by volume reinfusion from the CPB reservoir. Arterial blood samples were obtained for the measurement of milrinone plasma concentrations and to determine the dose response curve. In all three milrinone groups, cardiac index and velocity of circumferential fiber shortening (Vcfc) significantly increased from the baseline, and both were significantly higher at 5 and 10 min than those in the control group. The plasma concentration of milrinone with half of maximum increase in Vcfc was 139.3 ng/mL based on the dose-response curve. Thus, milrinone improves hemodynamics and left ventricular function when constant loading conditions are maintained. PMID- 9212117 TI - Intracoronary levosimendan enhances contractile function of stunned myocardium. AB - A decrease in myofilament sensitivity to Ca2+ has been proposed as a mechanism for reversible contractile dysfunction after ischemia and reperfusion. The direct actions of intracoronary myofilament Ca2+ sensitizers on stunned myocardium have not been examined. Barbiturate-anesthetized dogs (n = 9) were instrumented for measurement of left ventricular (LV) and aortic blood pressure, cardiac output, left anterior descending coronary artery (LAD) blood flow velocity, and subendocardial segment length (percent segment shortening [%SS]). Dogs were subjected to five 5-min LAD occlusions interspersed by 5-min reperfusions. Three hours after the final reperfusion, levosimendan (1.5, 3, 6, and 12 microg/min) was administered via an intracoronary catheter. Hemodynamic effects and regional myocardial function were determined under control conditions, during each LAD occlusion and reperfusion, 3 h after final reperfusion, and after 10 min equilibration at each dose of levosimendan. Three hours after the final reperfusion, %SS and the ratio of effective to total regional work were significantly (P < 0.05) decreased, and postsystolic shortening area was increased, consistent with myocardial stunning. In stunned myocardium, intracoronary levosimendan caused dose-dependent increases in %SS (2 +/- 1 at 3 h after reperfusion to 13% +/- 2% during 12 microg/min), abolished postsystolic shortening area, and restored the ratio of effective to total regional work while producing minimum systemic hemodynamic effects. PMID- 9212118 TI - Effect of chronic and acute thyroid hormone reduction on perioperative outcome. PMID- 9212119 TI - A comparison of oral ketorolac and hydrocodone-acetaminophen for analgesia after ambulatory surgery: arthroscopy versus laparoscopic tubal ligation. AB - This multicenter study compared the analgesic efficacy and side effects of ketorolac and hydrocodone-acetaminophen when administered orally after ambulatory arthroscopic or laparoscopic tubal ligation procedures. After awakening from general anesthesia, 252 patients experiencing moderate or severe postoperative pain were randomly assigned to receive one of three analgesic treatments according to a placebo-controlled, double-blind protocol. Group 1 (n = 83) received oral ketorolac 10 mg every 6 h for up to 3 days, Group 2 (n = 82) received hydrocodone 7.5 mg plus acetaminophen 750 mg every 6 h for up to 3 days, and Group 3 (n = 87) received placebo capsules followed by ketorolac 10 mg every 6 h for up to 3 days. Severity of pain was recorded using a 4-point categorical score and visual analog scale (VAS) at 0.5 h and subsequently at hourly intervals for 6 h, as well as daily for up to 3 days. Pain relief was recorded using a 5 point categorical scale at the same time points. In the patients undergoing arthroscopic surgery, both ketorolac and hydromorphone-acetaminophen provided superior pain relief compared with the placebo. Although the categorical summed pain intensity difference (SPID), VAS SPID, and total pain relief scores were higher in the ketorolac group compared with the hydrocodone-acetaminophen group, the differences were not statistically significant. In the patients undergoing laparoscopic tubal ligation surgery, the three treatment groups displayed similar responses to the study medications. However, the ketorolac group scored higher in terms of overall tolerability than the hydrocodone-acetaminophen group. In conclusion, there was no difference in the efficacy between oral ketorolac and hydrocodone-acetaminophen combination in controlling pain after outpatient arthroscopic surgery procedures. Neither oral analgesic proved to be very effective after laparoscopic tubal ligation. PMID- 9212120 TI - The incidence of latex sensitivity in ambulatory surgical patients: a correlation of historical factors with positive serum immunoglobin E levels. AB - Increasing reports of latex-induced anaphylaxis make preoperative identification of latex-sensitive individuals an important concern. The incidence of latex sensitivity and the efficacy of questionnaires in identifying this in ambulatory surgical populations have not been determined. To clarify these issues, 996 ambulatory surgical patients were studied preoperatively. A questionnaire addressing demographic information, previous surgeries, history of atopy, previous exposure or reactions to latex, congenital abnormalities, and food allergies was administered. These data were then compared with serum anti-latex immunoglobin E (IgE) levels (AlaSTAT test), and risk factors, sensitivity, and specificity were determined. Of this population, 6.7% had IgE antibodies against latex (i.e., latex sensitivity). Male gender, non-Caucasian race, age, asthma, spinal cord abnormalities, food allergies, stated latex allergy, and symptoms when exposed to latex increased the risk of latex sensitivity. The specificity and positive predictive value of history were low. No systemic allergic reactions occurred, a finding that could be attributed to chance alone. The incidence of latex sensitivity in this population suggests that latex allergy is a significant potential problem in ambulatory surgical patients. History, however, does not appear to be a reliable predictor of the presence of anti-latex antibodies. PMID- 9212121 TI - Midazolam premedication delays recovery after propofol without modifying involuntary movements. AB - Midazolam has GABAergic effects in children that may modify propofol-induced involuntary movements, yet delay recovery. In a double-blind, randomized study, 24 children (2-7 yr of age, ASA physical status I or II) undergoing short surgical procedures received midazolam 0.5 mg/kg (Group M) or placebo (Group P) per os 20-30 min before propofol anesthesia (5 mg/kg intravenously followed by an infusion). Blind observers scored sedation and anxiety levels (scale 1-4) before premedication, at separation from parents, and at induction of anesthesia. Induction and emergence were videotaped, and body movements were recorded. During recovery, times to eye opening and maximum Steward (SS = 6) and Vancouver Sedative Recovery (VSRS = 22) scores were noted. Parents were questioned about side effects that may have occurred during the following week. Both groups were similar in age, sex, weight, timing of premedication, propofol dose, and duration of surgery. The incidence of involuntary movements did not differ between groups but was higher at induction (79%) than on emergence (25%) (P < 0.05). Anxiety and sedation scores were similar in Group P and Group M, but recovery took longer after midazolam, with eye opening (mean +/- SD) 24 +/- 7 vs 43 +/- 18 min, maximum SS (median and range) 27 (13-37) vs 55 (24-138) min, and maximum VSRS 51 (30-100) vs 80 (50-130) min. Children returned to normal activity in 1 (0-5) day, and none exhibited neurological complications. We conclude that an oral premedicant dose of midazolam prolongs recovery from anesthesia in children without affecting dystonic movements after propofol. PMID- 9212122 TI - Noninvasive monitoring of carbon dioxide during respiratory failure in toddlers and infants: end-tidal versus transcutaneous carbon dioxide. AB - We prospectively compared the accuracy of two noninvasive monitors of arterial CO2 (end-tidal and transcutaneous) in mechanically ventilated infants and toddlers with respiratory failure. The study included infants and toddlers less than 48 mo of age who required tracheal intubation and mechanical ventilation for respiratory failure. In each patient, both ETCO2 and transcutaneous CO2 (TC-CO2) were simultaneously monitored and compared with PaCO2 when an arterial blood gas analysis was performed. The cohort for the study included 25 toddlers and infants ranging in age from 1 to 40 mo and in weight from 3.3 to 19.1 kg. A total of 100 sample sets (PaCO2, ETCO2, TC-CO2) was compared. The ETCO2 to PaCO2 difference was 6.8 +/- 5.1 mm Hg, while the TC-CO2 to PaCO2 difference was 2.3 +/- 1.3 mm Hg (P < 0.0001). The absolute difference of the TC-CO2 and PaCO2 was 4 mm Hg or less in 96 of the 100 values, while the ETCO2 to PaCO2 difference was 4 mm Hg or less in 38 of the 100 values (P < 0.0001). Bland-Altman analysis revealed a bias of 0.68 with a precision of +/-2.35 when comparing the TC-CO2 and the PaCO2 and a bias of -6.68 with a precision of +/-5.01 when comparing ETCO2 with PaCO2. In neonates and infants with respiratory failure, TC-CO2 monitoring provided a more accurate estimation of PaCO2 than ETCO2 monitoring. PMID- 9212123 TI - Ventricular fibrillation during anesthetic induction in a child with undiagnosed mitral valve prolapse. PMID- 9212124 TI - Non-operating room emergency airway management and endotracheal intubation practices: a survey of anesthesiology program directors. AB - Airway management in the operating room is the responsibility of anesthesiologists, although a variety of personnel may be responsible for airway management outside the operating room. We conducted a survey of anesthesia program directors regarding emergency airway management practices at their institutions. A questionnaire was sent to anesthesia program directors listed in the Graduate Medical Education Directory for 1995-1996. Of the 153 programs surveyed, 134 (88%) responded. In 45% of institutions, intubations in the emergency ward (EW) were performed by emergency medical physicians, 32% by anesthesiology personnel, and 19% by both. Most intubations performed on the hospital ward were performed by anesthesiologists. Neuromuscular blocking drugs and sedative/hypnotics were used 90% and 95% of the time, respectively, by emergency medical physicians in hospitals in which they managed the airway independently. Our data serve as a snapshot of current practices. EW physicians are prominently involved in airway management in the emergency room both independently and with anesthesiologists. Airway management in trauma patients remains the domain of anesthesiologists. Anesthesiologists are most represented in airway management on hospital floors. PMID- 9212125 TI - Comparison of continuous spinal with combined spinal-epidural anesthesia using plain bupivacaine 0.5% in trauma patients. AB - We investigated the efficacy and complications of microcatheter spinal anesthesia (CSA) in comparison to a combined spinal-epidural technique (CSE) using plain bupivacaine 0.5%. Sixty trauma patients randomly received either CSA using a 22 gauge Sprotte needle and a 28-gauge microcatheter or CSE after insertion of a 22 gauge epidural catheter through an 18-gauge Tuohy needle followed by dural puncture with a 25-gauge pencil-point needle inserted through the backeye of the Tuohy needle. An initial subarachnoid bolus of 2 mL of plain bupivacaine 0.5% was injected. If analgesia did not reach T12 within 20 min, supplemental bupivacaine was injected either intrathecally or epidurally up to a maximum of 5 mL in the CSA group or 16 mL in the CSE group. Mean arterial blood pressure, heart rate, and analgesic levels were recorded. On postoperative Day 4, patients were interviewed for postanesthetic complaints. Technical problems were more frequent in the CSE group than in the CSA group (47% vs 13%). Performance of anesthesia was faster (8 +/- 3 vs 15 +/- 8 min) and the total dose of bupivacaine lower (3.2 +/- 1.0 vs 9.7 +/- 5 mL) in patients who received CSA. The incidence of hypotension did not differ significantly. However, more patients in the CSE group were treated for bradycardia (4 vs 0). The number of patients suffering from postdural puncture headache was comparable in both groups, but there were more patients with lower back pain in the CSE group (8 vs 2). In conclusion, our data suggest that microcatheter CSA is not associated with an increased rate of complication in patients with lower limb fractures. PMID- 9212126 TI - The effects of topical and intravenous ropivacaine on canine pial microcirculation. AB - To assess the direct cerebrovascular effects of ropivacaine, we studied pharmacological responses to its topical and intravenous (IV) administration on vasomotor tone of pial vessels in in vivo experiments using a parietal cranial window in 24 dogs anesthetized with pentobarbital. We directly measured the diameters of pial arteries and veins after the administration of five different concentrations of ropivacaine solution (10(-7) to 10(-3) mol/L) randomly given into the window (n = 10). In six dogs, after pretreating the pial vessels with yohimbine (10(-5) mol/L), the inhibitory action of yohimbine was examined after the application of ropivacaine (10(-3) mol/L). The effects of IV ropivacaine (1 and 4 mg/kg) were also evaluated in the remaining eight dogs. Ropivacaine produced significant constriction of the pial arteries in a concentration dependent manner (10(-7) to 10(-3) mol/L, P < 0.05) and only exerted a constrictive action on small veins (P < 0.05) at 10(-3) mol/L. Yohimbine had no effect on ropivacaine-induced constriction of pial vessels. IV ropivacaine, 4 mg/kg but not 1 mg/kg, caused pial vascular constriction (large arteries P < 0.005, small arteries P < 0.0001, large veins P < 0.01, small veins P < 0.005) associated with decrease in heart rate (P < 0.001). The results indicate that topical application of ropivacaine constricts pial arterial vessels in a concentration-dependent manner. A large dose of IV ropivacaine produced pial vasoconstriction associated with a decrease in heart rate and no decrease in mean arterial blood pressure. These effects do not appear to be mediated via the mechanism that depends on the activation of alpha2-adrenoceptors. We conclude that ropivacaine in high concentrations could, perhaps directly, cause significant constriction of the central nervous system vasculature. PMID- 9212127 TI - The effect of anesthetic techniques on blood coagulability in parturients as measured by thromboelastography. AB - Anesthetic techniques may affect blood coagulability and the subsequent incidence of thromboembolic events. The purpose of this study was to evaluate the effect of spinal and general anesthesia on blood coagulability in normal pregnant women undergoing cesarean section, using thromboelastography. In the spinal anesthesia group (n = 15), thromboelastography was performed after crystalloid preloading and during the immediate postanesthesia course. In the general anesthesia group (n = 15), thromboelastography was performed before induction and during the immediate postanesthesia course. Values for all thromboelastographic variables (reaction time [r], clot formation time [K], coagulation time [rK], maximum amplitude [MA], elastic shear modulus [G], clot formation rate [alpha angle], and coagulation index [CI]) in the preanesthesia period were similar in both the spinal and general anesthesia groups. However, in the postanesthesia period, r and K significantly decreased (P < 0.05), and alpha angle (P < 0.05) and CI significantly increased (P < 0.01) in the general anesthesia group when compared with the spinal anesthesia group. In the postanesthesia period, MA and G were similar in both groups. In the spinal anesthesia group, thromboelastographic variables did not change significantly in the postanesthesia compared with the preanesthesia period. We conclude that the use of general anesthesia for cesarean section is associated with accelerated coagulability when compared with spinal anesthesia. PMID- 9212128 TI - Comparative pharmacokinetics of ropivacaine and bupivacaine in nonpregnant and pregnant ewes. AB - We determined the pharmacokinetics and protein binding of ropivacaine and bupivacaine after intravenous administration to nonpregnant and pregnant sheep. All animals were in good condition throughout the study. The highest mean total serum drug concentrations were found at the end of infusion. For both drugs, pregnancy was associated with lower volumes of distribution during the terminal phase of drug elimination (V(d)beta) and steady state (V(d)ss), as well as with a lower total body clearance (CL). The relationship between V(d)beta and CL was such that the elimination half-life (T(1/2)beta) was not altered. There were also differences between the two drugs. In all animals, the distribution half-life (T(1/2)alpha), T(1/2)beta, volume of central compartment (V(c)), V(d)beta, V(d)ss, and mean residence times (MRT) were greater and CL lower for bupivacaine than ropivacaine. For both drugs, protein binding was concentration-dependent and greater in pregnant ewes. In conclusion, the pharmacokinetics of ropivacaine and bupivacaine are altered by ovine pregnancy in a similar way. If these data are applicable to humans, an unintended intravascular injection of either drug could be expected to result in higher total serum concentrations in the pregnant than in the nonpregnant patient, but drug levels would decline at similar rates in both groups of individuals. However, differences between the two drugs, particularly in T(1/2)beta and MRT, may make ropivacaine preferable for use in obstetric anesthesia. PMID- 9212129 TI - Thromboelastographic changes in healthy parturients and postpartum women. AB - Thromboelastography (TEG) using disposable plastic cups and pins was performed with native whole blood (native group) in 17 nonpregnant volunteers, 134 healthy term pregnant women (>36 wk gestation), and 69 postpartum women. Thromboelastography was also performed with celite-activated whole blood (celite group) in 15 nonpregnant female volunteers, 38 healthy term pregnant women, and 34 postpartum women. The thromboelastographic parameters r and K were significantly decreased in pregnant and postpartum women compared with nonpregnant women in both groups (P < 0.05). The maximum amplitude MA, elastic shear modulus, and alpha angles were significantly increased in pregnant and postpartum women compared with nonpregnant women in both groups (P < 0.05). The TEG coagulation index was significantly greater in pregnant and postpartum women compared with nonpregnant women in both groups. In this study, TEG showed that pregnancy is a hypercoagulable state and that postpartum women remain hypercoagulable through the first 24 h postdelivery. The use of celite in TEG accelerated the speed of TEG analysis. PMID- 9212130 TI - Hemodynamic response and change in organ blood volume during spinal anesthesia in elderly men with cardiac disease. AB - Aging and disease may make the elderly patient with cardiac disease particularly susceptible to hypotension during spinal anesthesia. We studied 15 men, 59-80 y old, with histories of prior myocardial infarction (n = 9), congestive heart failure (n = 2), and/or stable myocardial ischemia (n = 11) given spinal anesthesia with 50 mg lidocaine in dextrose. Technetium-99m-labeled red blood cell imaging estimated left ventricular ejection fraction (EF) and changes in blood volume in the abdominal organs and legs. Arterial and pulmonary artery catheters provided hemodynamic measurements. Sensory block averaged T4 (range T1 10). Mean arterial pressure decreased 33% +/- 15% (SD) (P < 0.001), secondary to decreases in vascular resistance (SVR), -26% +/- 13% (P < 0.001) and cardiac output, -10% +/- 16% (P = 0.03). EF increased from 53% +/- 11% to 58% +/- 14% (P < 0.001) while left ventricular end-diastolic volume (LVEDV) decreased (-19% +/- 9%, P < 0.001). Blood volume increased in the legs (6% +/- 6%, P = 0.006), kidneys (10% +/- 9%, P < 0.001), and mesentery (7% +/- 5%, P 0.001) but not in the liver or spleen. Cardiac function was well maintained. We concluded that the primary mechanism of hypotension was a decrease in SVR, not cardiac output, despite the decrease in LVEDV. PMID- 9212131 TI - Spinal conduction block by intrathecal ketamine in dogs. AB - In addition to its use for intravenous (I.V.) anesthesia, ketamine can provide pain relief in humans when administered spinally. To elucidate the mechanisms of intrathecal (I.T.) ketamine analgesia, we observed differences in the effects of I.V. and I.T. ketamine on intraspinal evoked potentials (ISEPs) in 28 dogs anesthetized with pentobarbital. Bipolar extradural electrodes were inserted at the cervical and lumbar regions of the spinal cord for recording descending ISEPs represented by the two negative deflections, Waves I and II. I.V. ketamine 2 and 10 mg/ kg did not affect the amplitude and latency of Wave I, whereas the large dose (10 mg/kg) significantly decreased the amplitude but not the latency of Wave II. I.T. ketamine 1 and 5 mg/kg caused significant dose-dependent decreases in both Wave I and II amplitudes and prolongations of both Wave I and II latencies. These I.T. effects on ISEPs are consistent with previous in vitro observations that ketamine blocks axonal conduction. We conclude that axonal conduction block may contribute to the analgesic mechanism of I.T. ketamine. PMID- 9212133 TI - Patient experience of pain after elective noncardiac surgery. AB - The purpose of this study was to examine the extent and evolution of pain after common major surgical procedures and to establish correlates of three types of pain: pain at rest, pain with movement, and maximum pain over the previous 24 h. Patients completed a preoperative questionnaire to obtain data on age, gender, narcotic use, baseline level of pain, chronicity of pain, and level of anxiety. Patients were then interviewed on Postoperative Days 1, 2, and 3 to assess their pain on a scale of 0 (none) to 10 (worst imaginable). The mean pain score at rest was 2.6 on Postoperative Day 1 and decreased to 2.3 on Postoperative Day 3 (P = 0.06). The mean pain score with movement was 4.5 on Postoperative Day 1, which decreased to 4.2 on Postoperative Day 3 (P = 0.03). The mean maximum pain score over the previous 24 h was 6.3, which decreased to 5.6 (P = 0.0001). Preoperative narcotic use and high baseline preoperative pain, defined as a score > or = 4, were significantly (P < 0.05) associated with increased pain at rest, pain with movement, and maximum pain. Epidural analgesia was the only mode of analgesia significantly associated with both decreased postoperative pain at rest and decreased pain with movement (P < 0.05). These relatively high pain scores and minimum decreases in pain from Postoperative Days 1 to 3 emphasizes the need for more effective pain management continuing into the postoperative period to facilitate mobilization and recovery. PMID- 9212132 TI - A novel supraclavicular approach to brachial plexus block. AB - We describe a novel supraclavicular approach to the brachial plexus. Designated as the intersternocleidomastoid technique, this new approach was tested in unembalmed cadavers. It was then applied for evaluation to 150 ASA grade I or II patients scheduled for elective surgery or physiotherapy of the upper limb or for treatment of reflex sympathetic dystrophy associated with painful shoulder. The new approach was easy to master because of a very simple surface landmark, i.e., the triangle formed by the sternocleidomastoid heads, which were visible and palpable in most patients studied (90%). The procedure was effective intraoperatively, providing satisfactory anesthesia in 140 patients (93%), partially satisfactory blocks in 6 (4%), and unsatisfactory blocks in only 4 (3%). The catheter entry point is cephalad enough not to obscure the surgical field on the shoulder. Catheter insertion was successful in 63 of 70 patients. Postoperative analgesia was provided for 48 h or more in 45 patients and for 24 h in 18 patients. Only minor complications were observed: asymptomatic phrenic nerve block in 89 patients (60%), transient Horner's syndrome in 15 (10%), transient recurrent laryngeal nerve blockade in 2, and misplacement of the catheter into the subclavian vein in 1 patient. No pneumothorax was observed. PMID- 9212134 TI - The effects of adding sufentanil to bupivacaine for postoperative patient controlled epidural analgesia. AB - We tested the hypothesis that postoperative patient-controlled epidural analgesia was more effective with the combination of sufentanil and bupivacaine (Group 2) than with bupivacaine alone (Group 1). One hundred patients undergoing thoracic, upper abdominal, and aortic surgery were provided with an epidural catheter and randomly allocated to one of the two groups. Postoperatively, patients were monitored in a postanesthetic care unit for at least 1 day before they were transferred to a ward. Both groups had similar demographics and operations. Pain treatment was continued for 4.4 +/- 0.6 and 4.5 +/- 0.7 days for Groups 1 and 2, respectively. Although Group 2 patients needed less volume of the epidural analgesics on Postoperative Days 1 and 2, they reported lower pain intensity at rest and during activity for the first three postoperative days. The groups did not differ from each other regarding the incidence of respiratory depression. There was no late respiratory depression; however, three cases of early respiratory depression were detected and easily treated (Group 1 one event, Group 2 two events). Motor block was only seen in patients with lumbar epidural catheters. There was no difference between groups, and all patients with thoracic catheters could be mobilized beginning on the first postoperative day. We conclude that 1) the addition of sufentanil to a small-dose bupivacaine augments epidural analgesia and 2) thoracic epidural catheters should be used for postoperative analgesia after abdominal or thoracic surgery. PMID- 9212135 TI - Management of patient-controlled analgesia: a comparison of primary surgeons and a dedicated pain service. AB - Although Patient-Controlled Analgesia (PCA) is routinely available in most hospitals in the United States, there appears to be little standardization regarding who provides this valuable service to postoperative patients. This study evaluates the differences in PCA management practices and patient outcomes between primary service (PS) physicians and acute pain service (APS) physicians. Over a 3-mo period, 40 patients prescribed PCA by PS physicians were prospectively studied without the knowledge of the physicians or nurses involved in PCA management. After collecting PS data, a proportionate stratified random sampling procedure was used to select 40 APS patients matched for gender, age, and type of surgery. Data regarding patient demographics, PCA prescription, changes in PCA orders, opioid consumption, reason for discontinuation of PCA, verbal analog scale pain scores, side effects, and post-PCA pain management were analyzed. Although pain scores were not different between groups, APS patients had fewer side effects, were more likely to receive a loading dose, had their PCA settings adjusted more often (P < 0.05), and used more opioid. PS patients were more likely to receive intramuscular medications after PCA discontinuation (P < 0.05). This study demonstrates potentially important PCA management differences between APS and PS physicians. PMID- 9212136 TI - Preemptive epidural morphine for postoperative pain relief after lumbar laminectomy. AB - This study was designed to evaluate the efficacy of preemptive epidural morphine for postoperative analgesia after lumbar laminectomy. Thirty ASA physical status I adults undergoing elective lumbar laminectomy under general anesthesia were randomly allocated to one of two groups. Group 1 (study group) received 3 mg epidural morphine preemptively 60 min before surgery, followed by epidural placebo at the end of surgery. Group 2 (control group) received epidural placebo at the same time preoperatively as the study group, followed by 3 mg epidural morphine at the conclusion of surgery. Pain was assessed using visual analog scales (VAS), and sedation was graded on a 4-point rank drowsiness score. Time to first postoperative analgesic (TFA), the supplementary analgesia, and the amount of morphine used over the 24-h period were noted for the groups. VAS pain scores were significantly less in Group 1 (preemptive group) than in Group 2 8 h after surgery (P < 0.05). TFA in the study group (19.9 +/- 2.3 h) was significantly prolonged compared with the control group (8.5 +/- 1.0 h, P < 0.05). The demand for supplementary analgesia and postoperative morphine consumption in the preemptive group was significantly lower than that in control group (P < 0.05). Patients in the control group were significantly sedated after 12 h and had a high incidence of nausea and vomiting (P < 0.05). The study shows that preemptive epidural morphine is superior to epidural morphine given postoperatively for pain relief after lumbar laminectomy. PMID- 9212137 TI - Laryngeal mask airway and the incidence of regurgitation during gynecological laparoscopies. AB - We studied the incidence of regurgitation in 100 patients undergoing elective gynecological laparoscopies under general anesthesia with intermittent positive pressure ventilation using a laryngeal mask airway (LMA). Patients ingested methylene blue capsules 10-15 min before induction of anesthesia. After induction and insertion of an LMA using the recommended insertion technique, a fiberoptic examination of the larynx was made for traces of dye and to site a pH probe in the bowl of the LMA for continuous monitoring. LMA insertion was successful in all patients within two attempts (95 at first attempt). Fiberoptic examination revealed the vocal cords or cords and posterior or anterior epiglottis in 96 and no trace of dye in 99 patients. One patient regurgitated dye immediately after induction, and the stain was seen on the LMA after removal. The remaining 99 LMAs were not stained. Thirty patients were randomly selected for fiberoptic examination of the laryngopharynx before neuromuscular block was antagonized. Methylene blue staining did not occur in any of these patients. In 91 patients with complete pH data, regurgitation (pH < 4.0) did not occur. The 95% confidence limit for a true probability of regurgitation in this study is 0.041 or a true rate of regurgitation of less than 4.1%. A larger study would be required to possibly demonstrate a lower incidence of regurgitation. This study confirms the clinical impression that the incidence of regurgitation during laparoscopies with a LMA is extremely low. PMID- 9212139 TI - Classification of malignant hyperthermia-equivocal patients by 4-chloro-M-cresol. AB - To clarify the contracture response to 4-chloro-m-cresol (4-CmC) in malignant hyperthermia (MH) equivocal (MHE) muscle, we studied the effect of cumulative concentrations of 4-CmC. In vitro contracture test (IVCT) was performed in 35 probands according to the European MH test protocol. Surplus muscle bundles were exposed to 4-CmC (25-200 micromol/L), maintaining each concentration for 4 and 8 min. After 4 min exposure, the contracture increase of MH susceptible (MHS) (n = 7) muscle specimens was significantly (P = 0.05) greater at 50 micromol/L compared with either MHE halothane sensitive (MHEh) (n = 13) or MH normal (MHN) (n = 15) classified patients. Statistically significant differences (P < 0.05) were also found at 75 micromol/L. Exposure for 8 min yielded significant differences at 50 micromol/L only between MHS and MHEh. MHEh muscles revealed a dose-response curve similar to that found in MHN specimens. MHS muscles showed a significantly higher sensitivity to 4-CmC than either MHEh or MHN, and, in the probands tested so far, MHEh and MHN muscles seem to identically respond to 4 CmC, which seems to indicate a normal response in MHEh probands, implying no MH susceptibility. Therefore, 4-CmC might reduce the frequency of MHEh diagnosis based on standard halothane-caffeine IVCT. However, since MHE individuals may also represent an aberrant genetic status, with MH causing defects linked to unknown mutations, it is premature to consider 4-CmC as a solution to the diagnostic uncertainty of the true status of MHE probands. Presently, 4-CmC may provide supplementary information for a more precise phenotypic categorization of MHE individuals. PMID- 9212138 TI - Uric acid excretion increases during propofol anesthesia. AB - We compared the effect of propofol with that of sevoflurane anesthesia on uric acid (UA) excretion in ASA physical status I and II patients with normal renal function. A propofol group (n = 11) received propofol-nitrous oxide-fentanyl after induction of anesthesia by propofol, while a sevoflurane group (n = 12) received sevoflurane-nitrous oxide-fentanyl after induction of anesthesia by thiamylal. UA, creatinine (Cr), and urea nitrogen concentrations in serum and urine were measured before induction of anesthesia, 1, 2, and 3 h after induction, and on Postoperative Day 1. N-acetyl-beta-D-glucosaminidase, beta2 microglobulin concentrations, and pH in urine were also examined. Plasma clearance of UA (CUA) and Cr (CCr) were calculated. The hourly concentration and excretion of urine UA were significantly higher than those of the sevoflurane group (P < 0.01). Significant correlations were noted between the hourly urine volume and UA concentration (r = 0.58, P < 0.01 for the propofol group; r = 0.51, P < 0.01 for the sevoflurane group). The CUA of the propofol group was significantly higher than that of the sevoflurane group (22.9 +/- 10.6 vs 5.9 +/- 3.4 mL/min, mean +/- SD, P < 0.05). There were no significant differences in other renal variables between the two groups. The present study demonstrated that the UA excretion increased during propofol anesthesia, while it remained stable during sevoflurane anesthesia. PMID- 9212140 TI - Capnography during jet ventilation for laryngoscopy. AB - Jet ventilation is often used during laryngoscopy to permit improved visualization of the larynx and to eliminate a potentially flammable endotracheal tube when laser surgery of the airway is performed. Observation of chest wall movement and blood gas analysis are the usual standards for assessing the adequacy of ventilation during jet ventilation. It is reasonable to hypothesize that measurement of end-tidal CO2 concentrations during jet ventilation can be used to assess the adequacy of ventilation during jet ventilation. To test this hypothesis, end-tidal CO2 concentrations were determined during mechanical ventilation through an endotracheal tube and during jet ventilation. At the time that each end-tidal measurement was obtained, a sample of arterial blood was also obtained for later blood gas analysis. For both mechanical ventilation and jet ventilation, well defined relationships between end-tidal CO2 and arterial CO2 tensions were obtained. However, the relationships are distinct: the difference in arterial to end-tidal CO2 tension during supraglottic jet ventilation at a conventional respiratory rate was found to be 13.4 +/- 6.8 mm Hg (mean +/- SD) compared with 5.7 +/- 5.2 mm Hg obtained during conventional ventilation through an endotracheal tube. PMID- 9212141 TI - Maturation decreases ethanol minimum alveolar anesthetic concentration in mice as previously demonstrated in rats: there is no species difference. AB - The potency of conventional inhaled anesthetics increases with maturation: the 50% effective dose (minimum alveolar anesthetic concentration [MAC]) for conventional inhaled anesthetics in the neonatal rat or human exceeds MAC in the young adult. This increase also applies to ethanol in rats tested using MAC as the measure of anesthesia. However, the converse appears to be true for studies in mice assessed with the righting reflex; that is, adult mice are six times more resistant than neonates to the effects of ethanol. These disparate findings imply that maturation in rats and mice may produce opposing changes in the quantity or sensitivity of one or more receptors that mediate the actions of anesthetics that lead to the anesthetic state. Such a finding would be important for two reasons. First, both rodents are widely used in studies of anesthetic effects, and, thus, a species-dependent divergence in anesthetic effects has immediate experimental implications. Second, confirmation of such a species difference would supply an opportunity to test which receptors might be crucial to anesthetic mechanisms. Accordingly, we investigated whether maturation decreased ethanol potency in mice, using MAC as the measure of anesthesia. Applying standard techniques, we tested MAC for ethanol in 15 CF-1 mice aged 10 days (6-8.5 g) and in 13 mice aged 77-84 days (34-39 g). MAC decreased with maturation, and the decrease was indistinguishable from that found in our previous studies of rats. PMID- 9212142 TI - Midlatency auditory evoked potentials predict movements during anesthesia with isoflurane or propofol. AB - To determine threshold values, sensitivity, and specificity of midlatency auditory evoked potentials (MLAEP) for prediction of spontaneous intraoperative movements, 40 patients undergoing elective laparotomy were studied. Continuous epidural analgesia was used in all patients. To maintain general anesthesia, the patients in Group 1 (n = 20) received isoflurane (0.4-1.2 vol%), and the patients in Group 2 (n = 20) received propofol (3-5 mg x kg(-1) x h(-1) intravenously). Spontaneous movements were documented intraoperatively. Auditory evoked potentials were recorded continuously until the end of anesthesia. Latencies of the peaks V, Na, Pa, Nb, and P1 (ms) and amplitudes Na/Pa, Pa/Nb, and Nb/P1 (microV) were measured. Changes of MLAEP latencies and amplitudes during anesthesia were similar in both groups. Anesthesia led to statistically significant increases in the latencies of Na, Pa, Nb, and P1 and decreases in the amplitudes of Na/Pa, Pa/Nb, and Nb/P1 compared with the awake state. Before and during spontaneous movement observed intraoperatively or during emergence from anesthesia, the latencies of the peaks Na, Pa, Nb, and P1 decreased, and the amplitudes Na/Pa, Pa/Nb, Nb/P1 increased significantly. A threshold value of 60 ms of Nb proved to be most predictive of movement during anesthesia. MLAEP recording seems to be a promising method to monitor the level of anesthesia as defined by spontaneous movement during anesthesia. PMID- 9212143 TI - Interaction of midazolam with the nicotinic acetylcholine receptor of mouse myotubes. AB - The effects of midazolam on the peripheral embryonic nicotinergic acetylcholine receptor (nAChR) of mouse myotubes were studied to elucidate the mechanism of its effect on neuromuscular transmission. Standard patch clamp techniques on outside out patches were used. Pulses of 10(-4) M acetylcholine (ACh) applied by a liquid filament switch technique elicited macroscopic channel currents with a peak current amplitude of approximately 40 pA within <1 ms. The current decayed with a time constant of 30-100 ms due to desensitization. When midazolam was added in stepwise increased concentrations (10(-7) M to 7 x 10(-4) M) to the pulses, the current decay became bi-exponential, and a concentration-dependent decrease of the fast component of decay was observed. The current amplitude, however, was reduced slightly, and only at high concentrations of midazolam. This may indicate that midazolam binds to the open channel to cause the block. The rate constant of block (b(+1)) was found to be 1.8 x 10(6) M/s. Recovery experiments revealed a rate of unblocking (b(-1)) of approximately 2 x 10(-1) s(-1). After preincubation of the patches with midazolam, a substantial reduction of the current amplitude was seen at very low midazolam concentrations (<10(-7) M), which suggests an additional closed channel block with a Kd of approximately 10(-6) M. This closed channel block may be responsible for the muscle-relaxing effects of midazolam. PMID- 9212144 TI - Effect of midazolam on development of acute tolerance to alfentanil: the role of pharmacokinetic interactions. AB - This study in rats was performed to explore whether the inhibitory effect of midazolam on the development of acute tolerance to the analgesic effect of alfentanil is due to pharmacokinetic mechanisms. Analgesia was determined with tail-compression and hot-plate tests. Alfentanil and midazolam concentrations in plasma and the brain were measured using a radioimmunoassay and chromatographic technique, respectively. After the 4-h period of alfentanil administration (155 microg x kg(-1) x h(-1) after a 50-microg/kg bolus), when acute tolerance had already developed, midazolam (2-mg/kg bolus) enhanced the alfentanil-induced analgesia by 120% (P < 0.001) with the tail-compression test and 76% (P < 0.01) with the hot-plate test. At the height of midazolam-induced enhancement of the analgesic effect of alfentanil, the measurements of the alfentanil and midazolam plasma and brain concentrations did not demonstrate any significant changes in the drugs' concentrations. The results confirm that midazolam attenuates the development of acute tolerance to the analgesic effect of alfentanil and indicate that this interaction is not pharmacokinetic. PMID- 9212145 TI - Minimum alveolar anesthetic concentration values for the enantiomers of isoflurane differ minimally. AB - Results of in vivo and in vitro studies of the anesthetic potencies of the enantiomers (optical isomers) of isoflurane provide various results ranging from no difference to differences of nearly two fold. A finding of a difference in anesthetic requirement in the whole animal has particular relevance to theories of anesthetic mechanisms of action because it suggests that anesthesia may result from a specific anesthetic-receptor interaction. This led to our decision to redetermine the minimum alveolar anesthetic concentration (MAC) of (+)-S and (-) R enantiomers of isoflurane in 12 Sprague-Dawley rats (six per group). The (+)-S enantiomer gave a MAC of 0.0144 +/- 0.0012 atm (i.e., 1.44% +/- 0.12% at 1 atm pressure; mean +/- SD) and the (-)-R enantiomer gave a MAC of 0.0169 +/- 0.0020 atm. Although the 17% greater value for the (-)-R enantiomer is qualitatively consistent with previous results the difference is not significant (P = 0.06), and the absolute difference is smaller than that found by a previous study. However, given the small sample size, our power to define a small significant difference is limited. Regardless of statistical significance, our results do not confirm the conclusion that interaction with a specific receptor is important to the mechanism of action of inhaled anesthetics. PMID- 9212146 TI - Intravenous clonidine decreases minimum end-tidal isoflurane for induction of electroencephalographic burst suppression. AB - The aim of this study was to determine the individual end-tidal isoflurane (ET ISO) threshold concentration for the induction of electroencephalographic (EEG) burst suppression with and without intravenous (I.V.) clonidine and to evaluate the EEG and cardiovascular response to skin incision during isoflurane/N2O anesthesia. Thirty-nine patients (ASA physical status I or II, 20-68 yr of age) undergoing orthopedic surgery were randomly assigned to receive I.V. saline (n = 20) or I.V. clonidine (3 microg/kg, n = 19). After detection of isoflurane induced burst suppression, ET ISO was decreased in 0.1% ET steps until burst suppression diminished. Median minimum ET ISO for induction of burst suppression was 1.4% in the saline group and 0.9% in the clonidine group (P < 0.05). Before skin incision, EEG alpha 2 activity was significantly higher in the clonidine group compared with saline group. Fourteen patients (70%) in the saline group and 12 patients (63%) in the clonidine group showed a cardiovascular response to skin incision. After skin incision, EEG alpha 2 power was significantly decreased in both groups. A significant increase of delta activity was only found in the saline group. We conclude that the known minimum alveolar anesthetic concentration reduction of clonidine seems to be due to a direct cerebral action. PMID- 9212147 TI - Halothane inhalation inhibits the metabolism of chlorzoxazone, a substrate for CYP2E1, in rabbits. AB - We demonstrated the inhibitory effect of halothane (HAL) inhalation on the metabolism of chlorzoxazone (CZZ), a substrate for CYP2E1, in a bolus and a continuous injection study in rabbits receiving artificial ventilation. In a bolus injection study, the inhalation of 1.0% HAL significantly increased the half-life and the area under the curve and decreased the clearance of CZZ compared with those variables in midazolam administration. In a continuous injection study, the effect of various concentrations of inhaled HAL on plasma CZZ concentration at steady state was compared. Systolic and diastolic arterial blood pressure were decreased dose-dependently after 0.5%, 1.0%, or 2.0% HAL was inhaled. Although the plasma concentration of CZZ was increased 2.5-fold after 3 h of HAL inhalation, there was no significant difference in mean plasma concentrations among the groups treated with 0.5%, 1.0%, or 2.0% HAL. In contrast, the plasma concentration of lidocaine, a substrate for CYP3A, remained unchanged after 1.0% HAL was inhaled. These results suggest that general anesthesia obtained with HAL inhalation will affect the metabolism of drugs administered concomitantly when the drug is a substrate for CYP2E1. PMID- 9212148 TI - A concept for assessing interactions of general anesthetics. PMID- 9212149 TI - Does midazolam cause retrograde amnesia, and can flumazenil reverse that amnesia? PMID- 9212150 TI - Treatment of a paradoxical reaction to midazolam with haloperidol. PMID- 9212151 TI - Difficult airway management in a patient with traumatic asphyxia. PMID- 9212152 TI - Airway obstruction due to a Sengstaken-Blakemore tube. PMID- 9212154 TI - Subclavian vein compression and thrombosis presenting as upper extremity pain. PMID- 9212153 TI - Regional anesthesia for thyroidectomy in two patients with amiodarone-induced hyperthyroidism. PMID- 9212155 TI - Does normovolemic hemodilution decrease myocardial oxygen consumption despite increased heart work? PMID- 9212156 TI - The effect of intravenous lidocaine on somatosensory evoked potentials during scoliosis surgery. PMID- 9212157 TI - Propofol and early extubation after cardiac surgery. PMID- 9212158 TI - The safety of dantrolene in a patient with a severe cardiomyopathy requiring a heart transplant. PMID- 9212160 TI - Clinical implications of ischemic preconditioning. PMID- 9212159 TI - Broken small-gauge spinal needle. PMID- 9212161 TI - Cloning, expression, and chromosomal localization to 11p12-13 of a human LIM/HOMEOBOX gene, hLim-1. AB - We have identified a putative transcription factor, designated hLim-1, from human fetal brain using degenerate polymerase chain reaction (PCR) and cDNA library screening. The deduced open reading frame, derived from sequencing a 3.0-kb hLim 1 cDNA, encodes a protein of 384 amino acids with two cysteine-rich LIM domains and one homeobox (HOX) DNA-binding domain. The nucleotide sequence of hLim-1 cDNA is 87% identical to mouse Lim-1 and the predicted amino acid sequence is greater than 97% conserved. Expression patterns of hLim-1 were evaluated by Northern analysis and reverse transcription (RT)-PCR coupled with Southern blotting. HLim 1 expression was observed in human brain, thymus, and tonsillar tissue. Expression of hLim-1 was also observed in 58% of acute myelogenous leukemia (AML) cell lines and in four of five primary samples from patients with chronic myeloid leukemia (CML) in myeloid blast transformation. The gene encoding hLim-1 was mapped using fluorescence in situ hybridization (FISH) to human chromosome 11p12 13. The expression pattern and structural characteristics of the hLim-1 gene suggest that it encodes a transcriptional regulatory protein involved in the control of differentiation and development of neural and lymphoid cells. Its expression in CML in blast crisis suggests that it may be involved with progression in this disease; a prospective study is required to confirm this. PMID- 9212162 TI - Molecular cloning of htra2-beta-1 and htra2-beta-2, two human homologs of tra-2 generated by alternative splicing. AB - A yeast two-hybrid screen was performed to find new factors involved in pre-mRNA splicing. Using SC35 as a bait, we isolated a human cDNA bearing high homology to the Drosophila transformer-2 (TRA-2) protein. This cDNA was named htra2-beta1. htra2-beta1 is a nuclear protein that colocalizes with SC35 in a speckled pattern. It interacts with several SR proteins tested in yeast. A second form named htra2-beta2 is generated by alternative splicing. This isoform gives rise to a truncated protein without an SR domain. Both isoforms are evenly distributed throughout adult rat tissue. The ratio of these two isoforms changes after stimulation of primary human T-cell and primary rat spleen cell cultures, indicating that alternative splicing is involved in regulation of htra2-beta activity. PMID- 9212163 TI - Cloning and expression of two human lysophosphatidic acid acyltransferase cDNAs that enhance cytokine-induced signaling responses in cells. AB - Lysophosphatidic acid (LPA) and phosphatidic acid (PA) are two phospholipids involved in signal transduction and in lipid biosynthesis in cells. LPA acyltransferase (LPAAT), also known as 1-acyl sn-glycerol-3-phosphate acetyltransferase (EC 2.3.1.51), catalyzes the conversion of LPA to PA. In this study, we describe the isolation and characterization of two human cDNAs that encode proteins possessing LPAAT activities. These two proteins, designated here as LPAAT-alpha and LPAAT-beta, contain extensive sequence sequence similarities to microbial or plant LPAAT sequences. LPAAT-alpha mRNA was detected in all tissues with highest expression in skeletal muscle whereas LPAAT-beta was expressed predominantly in heart and liver tissues. Expression of these two cDNAs in an Escherichia coli strain with a mutated LPAAT gene (plsC) complements its growth defect and shifts the equilibrium of cellular lipid content from LPA to PA and other lipids. Overexpression of these two cDNAs in mammalian cells leads to increased LPAAT activity in cell-free extracts using an in vitro assay that measures the conversion of fluorescently labeled LPA to PA. This increase in LPAAT activity correlates with enhancement of transcription and synthesis of tumor necrosis factor-alpha and interleukin-6 from cells upon stimulation with interleukin-1beta, suggesting LPAAT overexpression may amplify cellular signaling responses from cytokines. PMID- 9212164 TI - Transcriptional induction of the agp/ebp (c/ebp beta) gene by hepatocyte growth factor. AB - Hepatocyte growth factor (HGF) is a pleiotropic factor with mitogenic, morphogenic, motogenic, cytotoxic, or growth inhibitory activity. Although the signaling of HGF is mediated through the cell membrane receptor c-Met, the molecular mechanism of downstream signal transduction remains obscure. In this report, we present evidence that shows HGF can stimulate the expression of AGP/EBP (C/EBP beta) and NF-kappaB, which are both key transcription factors responsible for the regulation of many genes under stress conditions or during the acute-phase response. Biochemical and functional analysis indicates that the HGF-responsive element is located in the region -376 to -352 (URE1) of the 5' upstream regulatory sequence of agp/ebp. Activation of NF-kappaB by HGF was observed to precede the induction of agp/ebp. Further studies indicate that NF kappaB can cooperate with AGP/EBP or other members of the C/EBP family to activate the agp/ebp gene in both URE1 and URE2-dependent manner. These results suggest that the induction of the agp/ebp gene by HGF is mediated at least in part by its activation of NF-kappaB. The activated NF-kappaB then interacts with AGP/EBP, resulting in the induction of agp/ebp. PMID- 9212165 TI - In vivo protein-DNA interactions on a glucocorticoid response unit of a liver specific gene: hormone-induced transcription factor binding to constitutively open chromatin. AB - Transcription from the liver promoter of a 6-phosphofructo-2-kinase/fructose-2,6 bisphosphatase (PFK-2) gene depends on the presence of glucocorticoids that act via a glucocorticoid response unit (GRU) located in the first intron. The promoter and the GRU are in a constitutively open chromatin configuration. To determine how glucocorticoids would affect factor binding to the GRU in absence of chromatin remodeling, we have used a combination of in vitro DNA-binding assays and in vivo genomic footprinting in rat hepatocytes and hepatoma cells. We found that, in the absence of glucocorticoids, the GRU binds nuclear factor-I (NF I). Glucocorticoid treatment modified factor binding to the NF-I site and induced the binding of hepatocyte nuclear factor-3 (HNF-3). Transfection assays showed that HNF-3 cooperates with the glucocorticoid receptor in stimulating transcription. In contrast with the lack of effect of glucocorticoids on factor binding to constitutively open GRUs of other genes, HNF-3 binding to the open PFK 2 GRU was hormone-dependent. Therefore, the PFK-2 GRU behaves as a novel type of GRU. PMID- 9212166 TI - Regulation of the hepatic CYP 2E1 gene during chronic alcohol exposure: lack of an ethanol response element in the proximal 5'-flanking sequence. AB - Chronic exposure to ethanol is known to cause a dramatic increase in the level of CYP 2E1 apoprotein. More recently it has been demonstrated that under certain conditions the mRNA encoding cytochrome P450 2E1(CYP 2E1) is inducible; however, the mechanisms by which these increases occur are not well understood. In the current study, DNase I footprinting assays performed on the first kilobase of the CYP 2E1 5'-flanking sequences resulted in the identification of 13 sequence specific protected regions using rat liver nuclear extracts isolated from either control or ethanol-treated animals. No differences were observed in the DNase I footprint patterns produced by the two different nuclear extracts. In addition, analysis by electrophoretic mobility shift assays (EMSA) revealed that with one exception, there were no differences in the level of binding complexes between the two extracts. However, EMSA analysis with an oligonucleotide to one footprint site (designated Site C) revealed that in nuclear extracts isolated from ethanol treated animals there was a 2.9-fold increase in this binding complex when compared to control nuclear extracts. This site was previously shown to contain an HNF-1alpha binding site, and here we demonstrate that bacterially expressed HNF-1alpha in footprint assays bind Site C sequences and that HNF-1alpha transactivates the CYP 2E1 promoter in co-transfection experiments with HNF 1alpha expression plasmid and plasmids containing CYP 2E1 promoter sequences coupled to the chloramphenicol acetyl transferase gene. Furthermore, in contrast to the increase observed by EMSA in Site C binding, no increase was detected in the CYP 2E1 transcriptional rate supported by nuclear extracts from ethanol treated animals over controls using in vitro transcription assays, suggesting that the increase by ethanol in CYP 2E1 transcription is not mediated through the HNF-1alpha site. PMID- 9212167 TI - Cellular targets for activation by c-Myc include the DNA metabolism enzyme thymidine kinase. AB - Although a remarkable number of genes has been identified that are either activated or repressed via c-Myc, only few of them obviously contribute to Myc's biological effect--the induction of proliferation. We found that in logarithmically growing cells overexpression of Myc specifically induces thymidine kinase (TK) mRNA expression and enzyme activity, whereas loss of one allele of Myc causes downregulation of this enzyme. We show that activation of Myc triggers high levels of this normally strictly S-phase-regulated DNA metabolism enzyme in serum arrested G0 cells and causes high and constant levels of TK expression throughout the entire ongoing cell cycle. Induction of TK by Myc requires an intact transcriptional activation domain. Myc-induced deregulation of this enzyme is paralleled by alterations of protein binding at the E2F-site of the TK promoter. We further show that cell growth arrest by the cyclin-dependent kinase inhibitor p16 is abrogated by overexpression of Myc and that co overexpression of p16 cannot inhibit the Myc-induced up-regulation of TK expression. Our data demonstrate TK to be a cellular target of Myc independently of the status of cell proliferation and provide evidence that the transcription factor E2F might be involved in this process. PMID- 9212168 TI - Cystic fibrosis gene mutation (deltaF508) is associated with an intrinsic abnormality in Ca2+-induced arachidonic acid release by epithelial cells. AB - The mechanism(s) of chronic airway inflammation in cystic fibrosis (CF) remains poorly understood. We studied Ca2+-induced release of arachidonic acid (AA), a precursor of proinflammatory lipid mediators, in epithelial cell lines with the deltaF508 mutation in CF transmembrane conductance regulator (CFTR) gene and in those lacking this mutation or cells in which this mutation was corrected by a functional CFTR gene transfer. We found that: (i) the mutant cells manifested an abnormally high Ca2+-induced AA release as compared to controls, (ii) AA release appeared to be catalyzed by a phospholipase A2 (PLA2) but not by phospholipase C followed by diacylglycerol lipase, and (iii) either correction of the CFTR mutation or inhibition of PLA2 activity rectified this AA release abnormality. Taken together, our results suggest that CFTR mutation is associated with an intrinsic abnormality in AA release by epithelial cells carrying the deltaF508 mutation and suggest that the mechanism of chronic airway inflammation in CF, at least in part, involves this abnormality. These results also partly explain the effectiveness of high-dose ibuprofen therapy in arresting the progression of destructive lung disease in CF. Furthermore, they raise the possibility that correction of abnormal AA release by inhibiting PLA2 activity may improve the therapeutic benefits of ibuprofen. PMID- 9212170 TI - Closely related genes encode developmental and tissue isoforms of porcine cytochrome P450 aromatase. AB - We examined the chromosomal basis for the synthesis of tissue (ovary, endometrium/placenta, and peri-implantation blastocyst) isoforms of cytochrome P450 aromatase in the pig. DNA fragments derived from three distinct porcine aromatase chromosomal genes were cloned and characterized. The porcine type III aromatase gene encoding the blastocyst aromatase isoform was found to consist of nine coding exons and two mutually exclusive, 5' untranslated exons (designated E1A and E1B), collectively spanning 30 kb or more. The porcine type II aromatase gene, encoding the endometrial/placental aromatase isoform, was identified by cloning of a genomic DNA fragment spanning the corresponding exons 7, 8, and 9. The DNA inserts of two other phage clones encompassed exons 2, 3, and 4 of a third chromosomal gene (type I) encoding the ovarian aromatase isoform. All intron-exon junctions in these genomic fragments were found to be identical in relative positions to those of the single-copy human aromatase gene. Comparisons of cDNA and genomic sequences indicated that nucleotide sequence variation was not uniform across the corresponding exons of these genes and that the corresponding intronic sequences were conserved. The type II and type III aromatase genes were localized to the same regional location (q16-17) on swine chromosome 1, which is homologous to the human chromosome 15 region (q21.1) in which the human aromatase gene resides. Results demonstrate that the three aromatase genes characterized in the present study appear to be similar in their overall structural organization and most likely are clustered, which could have resulted from at least two independent gene duplication events. The presence of multiple aromatase genes constitutes a newly described mechanism by which aromatase enzyme biosynthesis and functional activity can be regulated in a tissue and temporal fashion and serves to highlight further the complexity of aromatase gene expression in mammals. Moreover, the presence of a unique aromatase gene that is highly expressed in pig blastocysts may constitute a paradigm for other mammals (e.g., equids, rabbit, hamster) whose peri implantation blastocysts are estrogenic. PMID- 9212169 TI - Enhanced expression in spleen macrophages of the mouse homolog to the human putative tumor suppressor gene ZFM1. AB - We have characterized the cDNA of MZFM, the mouse homolog to the novel human putative tumor suppressor gene ZFM1. The total length of the cDNA is 2,637 nucleotides with an open reading frame for a protein of 548 amino acids containing 4.7% methionine and 17.2% proline. The predicted molecular mass of 59 kD fits the 62-kD band experimentally determined by NaDodSO4-PAGE from in vitro translation products of in vitro-transcribed MZFM cDNA. The MZFM cDNA best matches to that ZFM1-isoform without the so-called 0.25-kb E-domain and to the L49345 cDNA recently identified in a human leukemia cell line. Northern analysis reveals expression of MZFM only in spleen macrophages. Reverse transcription polymerase chain reaction (RT-PCR) in combination with Southern analysis also detects a low basal expression in splenic T cells and B cells, as well as in other tissues such as heart, kidney, brain, liver, testis, bone marrow, adrenal gland, lymph nodes, pancreas, and thymus. In splenic macrophages, MZFM mRNA is alternatively spliced yielding a 3.6-kb transcript with E-domain, a 3.0-kb transcript without E-domain, and a 2.7-kb transcript with E-domain. The predicted MZFM protein contains diverse functional domains, i.e., a nuclear localization signal, a metal binding motif, a glutamine/proline stretch, proline-clusters, a CGA-motif, and a QUA1-KH-QUA2 region, thus indicating multiple functions of MZFM. Presumably, MZFM is a new member of those proteins combining features of signal transduction and RNA activation (STAR-proteins). The different MZFM-isoforms may be part of a macrophage-inherent program of transduction of environmental signals into different activational states of macrophages. PMID- 9212171 TI - Identification of essential cis-acting regulatory elements for transcription of the rat DAN gene. AB - The DAN gene was initially isolated as one of the genes whose expression is significantly decreased in a variety of transformed rat fibroblasts 3Y1 cells when compared with the parental 3Y1 cells. In the present study, we have isolated the genomic clone of the DAN gene from a 3Y1 genomic library and characterized the possible regulatory elements responsible for the transcription of the DAN gene. The transcription initiation site was determined by a primer extension experiment. Putative TATA and CAAT-like elements were present 31 and 358 bp upstream from the transcription start site, respectively. Transient transfection of a series of DAN-chloramphenicol acetyltransferase (CAT) gene constructs, which contain different portions of the 5'-flanking region (2,236 bp) of the DAN gene and the CAT gene, was used to localize a regulatory element. These experiments demonstrated the presence of the regions that regulate DAN gene expression positively (-57 to +118) and negatively (-1,232 to -636). The electrophoretic mobility-shift assays revealed that 3Y1 and SR-3Y1 nuclear extracts specifically interact with the positive (-57 to +118) and the negative (-1,226 to -987) regulatory regions, respectively. PMID- 9212172 TI - Kinesin light chain in a eubacterium. AB - A eubacterial homolog of a kinesin light chain gene has been isolated and characterized from the cyanobacterium Plectonema boryanum. Although the eubacterial and eukaryotic kinesin light chains are highly similar in amino acid sequence, the eubacterial sequence differs in several distinguishing structural features, including the absence of a putative PEST domain and the presence of additional highly conserved imperfect tandem repeats. Two soluble kinesin light chain antigens have been identified from whole-cell lysates by immunoblot analysis. Attempts to identify a canonical kinesin heavy-chain gene or protein were unsuccessful, suggesting that a kinesin heavy chain may be absent or unnecessary for kinesin light-chain function in this eubacterium. Our findings establish that certain basal elements of eukaryotic cellular transport appear to be resident in eubacteria. We discuss the possibility that the eukaryotic kinesin light chain was acquired by lateral gene transfer. PMID- 9212173 TI - A simple improvement in expression cloning. AB - Expression cloning is an effective approach for isolating genes encoding proteins that associate with a target species. Several molecules have been isolated by expression cloning, including CRE-BP1 associating with Jun (Macgregor et al., 1990); Grb1, identical to p85 PI3-kinase, with the EGF receptor (Skolnik et al., 1991); and Max with Myc (Blackwood and Eisenman, 1991). Expression cloning involves induction of proteins from a lambda gt11 cDNA expression library and screening the proteins on nitrocellulose membranes using a peptide probe (Macgregor et al., 1990). With this method, we previously isolated an Lck tyrosine kinase-associated protein, LckBP1, which is identical to HS1 (Kitamura et al., 1989, 1995; Takemoto et al., 1995). In those experiments, we used a glutathione S-transferase (GST)-Lck SH3 domain fusion protein as a probe, followed by detection of the complex with anti-GST polyclonal antibody. Whereas the ease of obtaining the fusion construct and high-titer anti-GST polyclonal antibody represented clear advantages, the system suffered from high background and low sensitivity. Here we show that pretreatment of nitrocellulose filters with NaDodSO4 reduces background and, in turn, increases sensitivity. PMID- 9212174 TI - A novel tunicate (Botryllus schlosseri) putative C-type lectin features an immunoglobulin domain. AB - We have cloned a putative C-type lectin of Botryllus schlosseri [Ascidiacea], whose deduced protein of 333 amino acids features three building blocks: (i) a Greek-key motif signature at the amino-terminus, (ii) a C-type lectin domain signature, and (iii) an immunoglobulin (Ig) domain at the carboxyl terminus. This C-type lectin was termed BSCLT. Similarity searches revealed that the Ig domain in BSCLT, which is evidently not polymorphic, is best classified as an Intermediate-type Ig domain. Rabbit antibodies, raised against recombinant BSCLT, cross-reacted in a Western blot with a 38-kD polypeptide in tunicate crude extract. Presumably, this bimodal tunicate protein is the first description of a soluble lectin that features besides the carbohydrate recognition domain also a complete Ig domain. PMID- 9212175 TI - MN blood group affects response of serum LDL cholesterol level to a low fat diet. AB - Previous studies found that MZ twin pairs who are blood group NN have greater intrapair variability in plasma lipid levels than those who are MM or MN. This led to the prediction that the response of plasma lipid levels to a low fat diet would depend on MN blood group, the greatest response being in those who are NN. The present study was based upon 254 patients who took part in the Australian Polyp Prevention Project. This was a 2 x 2 x 2 randomised factorial design based upon the presence or absence of the three factors: a dietary fibre supplement, a beta-carotene supplement and reduced intake of dietary fat. The lowering of plasma, low density lipoprotein (LDL) cholesterol, in response to a low fat diet was greatest in those who were NN and least in MN heterozygotes. Overall, a reduction in LDL level was observed in the 47% of the APPP population who were on a low fat diet and who were homozygous MM or NN. The result was consistent with a balanced polymorphism at or near the GLYA locus on chromosome 4 that influences the sensitivity of plasma lipid levels to dietary fluctuations in fat intake. PMID- 9212177 TI - High prevalence of a novel mutation in the exon 4 of the low-density lipoprotein receptor gene causing familial hypercholesterolemia in Belgium. AB - In a cohort of 70 unrelated patients living in Southern Belgium with autosomal dominantly inherited hypercholesterolemia, 11 had a hitherto undescribed mutation in exon 4. It consisted in a C-->A mutation at nucleotide 366, resulting in a stop codon at residue Cys122. This C122X mutation is expected to cause a class I receptor defect. The biochemical and clinical data collected from the patients carrying the mutation were consistent with a severe form of familial hypercholesterolemia (FH). Some differences between generations were noted. Amongst the C122X carriers, those born after 1926 had cardiovascular complications earlier than those born before 1926. This raises the possibility that changes in environmental factors during the course of the century have had an unfavorable impact on the prognosis of the disease. The mutation was found in 16% of the suspected FH patients and less frequently (less than 3% of suspected FH) in Northern Belgium. The haplotype of the chromosomes carrying the mutation was the same in all C122X families, but extensive genealogical studies failed to reveal a common ancestor. We conclude that C122X is an old and common cause of FH in Belgium. Screening for this mutation may be useful in the diagnosis of FH in Belgium. PMID- 9212176 TI - Analysis of 65 Turkish patients with congenital aplastic anemia (Fanconi anemia and non-Fanconi anemia): Hacettepe experience. AB - During the last 14 years, 65 unrelated patients were diagnosed as having constitutional aplastic anemia (CAA). In 52 of 65 patients the diepoxybutane (DEB) test was positive. Comparison of several hematological and clinical parameters in Fanconi anemia (FA) (DEB+) and non-Fanconi anemia (non-FA)(DEB ) patients disclosed no statistically significant differences. The study indicated that in Turkey there were no peculiarities in associated congenital abnormalities in FA and non-FA. The rate of consanguinity was 78% in FA and 46% in non-FA, suggesting that also among the non-FA group recessively inherited disorders are hidden. The mean age at diagnosis in FA was 7.7+/-4.4 (1.8-12) and in non-FA 7.8+/-3.8 (2-15) years. Nine out of 52 FA and five out of 13 non-FA patients died during the follow-up period. Five of the 52 FA patients developed malignancies, three of them had acute myeloblastic leukemia (AML), one a squamous cell carcinoma of the gingiva, and another a hepatocellular carcinoma. Peliosis hepatica occurred in three of the FA and one of the non-FA patients. A total of seven patients stayed in remission without any medication. The remaining 58 patients were given 2-5 mg/kg of oxymetholone and 5 mg prednisolone treatment. Because of sustained remission, oxymetholone therapy was terminated in four of the 45 FA and two of the 13 non-FA patients. Detailed examination of the pedigrees of all of patients indicated the presence of multiple congenital anomalies. In seven of 52 FA and one of 13 non-FA patients there was increased risk for AML and/or other cancers among family members. PMID- 9212178 TI - Linkage studies exclude the AT-V gene(s) from the translocation breakpoints in an AT-V patient. AB - An 8-year-old girl with severe microcephaly of prenatal onset, borderline intelligence, defects of skin pigmentation, deficiency of both humoral and cellular immunity, a normal serum alpha-fetoprotein level and hypersensitivity to ionizing irradiation is described. Spontaneous chromosomal breakage in lymphocytes together with the clinical presentation led to the diagnosis of ataxia telangiectasia variant (AT-V). In addition, the patient carried a constitutional translocation of paternal origin: 46,XX,t(3;7)(q12;q31.3) pat. In subsequent linkage and haplotype studies in 12 AT-V families with microsatellite markers from each of the translocation breakpoint regions, we could clearly exclude the localization of an AT-V gene to these regions. PMID- 9212179 TI - The prevalence of Usher syndrome and other retinal dystrophy-hearing impairment associations. AB - The study was undertaken to procure population-based prevalence data on the various types of Usher syndrome and other retinal dystrophy-hearing impairment associations. The medical files on 646 patients with a panretinal pigmentary dystrophy aged 20-49 years derived from the Danish Retinitis Pigmentosa (RP) register were scrutinised. The data were supplemented by a prior investigation on hearing ability in a part of the study population. After exclusion of patients with possibly extrinsic causes of hearing impairments, 118 patients, including 89 cases of Usher syndrome were allocated to one of five clinically defined groups. We calculated the following prevalence rates: Usher syndrome type I: 1.5/100,000, Usher syndrome type II: 2.2/100,000, and Usher syndrome type III: 0.1/100,000 corresponding to a 2:3 ratio between Usher syndrome type I and II. The overall prevalence rate of Usher syndrome was estimated to 5/100,000 in the Danish population, devoid of genetic isolates. The material comprised 11 cases with retinal dystrophy, hearing impairment, and additional syndromic features. Finally, 18 subjects with various retinal dystrophy-hearing impairment associations without syndromic features were identified, corresponding to a prevalence rate of 0.8/100,000. This group had a significant overrepresentation of X-linked RP, including two persons harboring a mutation in the retinitis pigmentosa GTP-ase regulator (RPGR) gene. PMID- 9212180 TI - Two novel missense mutations in the ATP-binding domain of the adrenoleukodystrophy gene: immunoblotting and immunocytological study of two patients. AB - Two novel missense mutations, 1939G to A (R518Q) and 2017A to G (Q544R) were identified in Japanese patients with adrenoleukodystrophy (ALD). They are located in exon 6, which encodes part of the putative adenosine triphosphate binding domain of ALD protein. The ALD protein carrying the R518Q mutation was undetectable in fibroblasts, by immunoblot and immunofluorescence analysis, while the Q544R mutation had no apparent effect on the stability and localization of the ALD protein, but is expected to affect its function. PMID- 9212181 TI - Lissencephaly associated with cerebellar hypoplasia and myoclonic epilepsy in a Bedouin kindred: a new syndrome? AB - Clinico-radiological assessment of three mentally retarded members of a large Bedouin kindred showed lissencephaly, spastic paraparesis, myoclonic epilepsy and cerebellar hypoplasia. It seems that the familial association of lissencephaly/myoclonic epilepsy/cerebellar hypoplasia represents a new entity. PMID- 9212182 TI - Molecular and clinical studies of three cases of female pseudohermaphroditism with caudal dysplasia suggest multiple etiologies. AB - Female pseudohermaphroditism with caudal dysplasia is a clinical entity in which normal-appearing male genitalia may occur in the apparent absence of testosterone or the sex-determining gene (SR Y). We have extended observations of two previously reported cases, and report a third case, which strongly suggests multiple etiologies. The first case was one of identical twins. The other identical twin did not show female pseudohermaphroditism with caudal dysplasia, but both patients had the rare birth defect of neonatal cataracts. We have explored skewed X-inactivation as a possible difference between the two twins, with a negative result. The second case had a deletion at 10q25.3-->ter. This is near the location of PAX2, and we searched for mutations in PAX2 in both this and the first case, with negative results. Neither patient had a scrotal raphe, suggesting that a failure of division of the cloacal membrane was an important step in their development of female pseudohermaphroditism. The final case is newly described and differed from the above two in the presence of a scrotal raphe and an elevated testosterone level. Although no source for the testosterone was found, this case suggests that the etiology in this patient was different and that the presence of a scrotal raphe can be used to distinguish between at least two etiologies. PMID- 9212183 TI - Pearson marrow pancreas syndrome: a molecular study and clinical management. AB - Human mitochondrial DNA (mt DNA) lesions can cause a heterogeneous group of mitochondrial degenerative disorders. We report on a 5-year-old patient suffering from the full-blown picture of Pearson syndrome. His symptoms started in the first year of life with failure to thrive, followed by chronic diarrhoea and lactic acidosis at 18 months of age. Analysis of mitochondrial DNA revealed large amounts of mt DNA molecules with a 2.7 kb deletion in all tissues examined. The diagnosis of Pearson syndrome was made initially in the absence of haematological disturbances. In the following months neutropenia, sideroblastic anaemia and hypoparathyroidism developed. Daily administration of dichloroacetate (DCA) and bicarbonate controls the lactic acidosis, while episodic treatments with filgastrim (Neupogen) reverse episodes of severe neutropenia. Calcium and vitamin D supplementation compensate for the hypoparathyroidism. Chronic administration of DCA and supportive treatment for a long period help to stabilize patients with multiorgan dysfunction. PMID- 9212184 TI - Polydactyly and fetal hydantoin syndrome: an additional component of the syndrome? AB - We report a 3-year-old girl with fetal hydantoin syndrome (FHS) whose mother had received phenytoin 600 mg/day throughout gestation. She had growth retardation, mental deficiency, craniofacial dysmorphism and iris colomobata specific to FHS. However, the patient did not have the distal phalangeal hypoplasia which is associated with FHS; instead, she had polydactyly of the right foot. This seems to be the first FHS case in the literature with polydactyly. PMID- 9212185 TI - Molecular studies of an X;Y translocation chromosome in a woman with deletion of the pseudoautosomal region but normal height. AB - A translocation chromosome in a woman with the karyotype 46,X,der(X)t(X;Y)(p22.3; q11.2) was investigated by FISH and STS analysis with molecular probes derived from the sex chromosomes. Due to the partial deletion of the short arm pseudoautosomal region (PAR1) from DXYS14 to DXYS147 in the translocation chromosome, the proband is hemizygous for the gene responsible for growth control (SS) located in this region, yet does not show growth retardation. Molecular analysis of the Yq arm of the translocation chromosome revealed the presence of markers DYS273 to DYS246 harboring the hypothesized growth control gene critical region (GCY) on Yq, thereby placing the deletion breakpoint between markers DYS11 and DYS273. These results suggest that the Y-specific growth gene GCY on Yq compensates for the missing growth gene SS on Xp22.3. PMID- 9212186 TI - Female pseudohermaphroditism due to classical 21-hydroxylase deficiency in a girl with Turner syndrome. AB - We report on a rare case of female pseudohermaphroditism due to classical 21 hydroxylase deficiency associated with Turner syndrome (45,X/46,XX). Difficulties in the management of both diseases are briefly discussed. We regard this rare combination as a coincidental occurrence. PMID- 9212187 TI - APO E polymorphism in Spanish and Moroccan populations. AB - Apolipoprotein E (apoE) gene polymorphism was analyzed by polymerase chain reaction in one Moroccan and six Spanish populations, a total of 660 individuals. No significant differences were observed between samples, and the mean relative frequencies (with 95% confidence intervals) found were 0.104 (0.069-0.139) for the epsilon4 allele, 0.855 (0.813-0.897) for epsilon3 and 0.041 (0.015-0.067) for epsilon2. Frequencies of the epsilon4 allele were low in comparison to Northern European populations, but similar to those reported for other South-European populations. The presence of a rare mutation, E2 Christchurch, in one Basque individual was confirmed by sequence analysis. PMID- 9212188 TI - Brachydactyly in a child with duplication-deficiency subsequent to t(15;20)(q25.2;p12.2)mat. Candidate regions on one or both chromosomes? AB - We report a child with a duplication-deficiency subsequent to t(15;20)(q25.2;p12.2), transmitted in at least 5 generations, who showed features of 15q- syndrome. We speculate that brachydactyly--most likely because of brachymesophalangism--is a feature of the phenotype of this chromosomal aberration and points to candidate gene(s) in this region. A similar brachydactyly was, however, reported with dup(20p1-pter). PMID- 9212189 TI - Short report on DNA markers at candidate loci. Two new polymorphisms in the BRCA 1 gene. PMID- 9212190 TI - Cell growth on cordierite: an approach to the identification of reliable supports for continuous-flow solid-bed reactors. AB - This study aimed to assess the biocompatibility of two cordierite ceramics (DF and Cord 1014), with similar chemical composition and different porosity, as a potential support for cell growth in a continuous-flow, solid-bed reactor. The Chinese hamster ovary (CHO) cell line transfected with HBV-DHFR recombinant plasmid was seeded on cordierite or polystyrene dishes and evaluated for cell growth and production of recombinant hepatitis B surface antigen. Proliferation of the CHO cells, in terms of cell number, was generally similar in polystyrene and Cord 1014 and always lower in DF. Flow cytometric analysis showed no difference in cell cycle distribution for cells grown on different supports, and showed a two-fold increase in percentage of debris for cells grown on DF than for those grown on Cord 1014 and polystyrene culture dishes. Moreover, the morphology of cells grown on Cord 1014 did not change during the experiment, and cells were well spread and organized. Finally, total recombinant hepatitis B surface antigen production was higher on Cord 1014 than on polystyrene and DF samples. Such evidence suggests that Cord 1014 could be a promising support for growing cells in a continuous-flow, solid-bed reactor. PMID- 9212191 TI - Apatite deposition on titanium surfaces--the role of albumin adsorption. AB - Titanium implant surfaces are known to spontaneously nucleate apatite layers when in contact with simulated body fluids. However, adsorption of proteins may influence the process of apatite layer formation. In this study the role of bovine serum albumin (BSA) adsorption in the process of apatite deposition on titanium substrates is investigated. Deposition of calcium phosphate was induced by immersing titanium substrates in a Hank's balanced salt solution (HBSS) for times ranging from 1 to 23 days. The resulting substrates were studied by scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), wettability measurements and electrochemical impedance determinations. All these methods indicate the presence of a calcium phosphate layer. The same procedure was repeated substituting HBSS with a solution of BSA in HBSS. Although SEM, EDS and electrochemical impedance spectra do not reveal the presence of an apatite layer, XPS analysis strongly indicates that the inhibition of apatite formation by BSA is only partial. The competition between BSA adsorption and apatite deposition seems to lead to a mixed film where the protein co-exists with calcium phosphate. Wettability studies suggest that this surface film is heterogeneous and porous, similar to the thicker films formed in albumin-free HBSS. PMID- 9212192 TI - Biomechanical characterization of osseointegration during healing: an experimental in vivo study in the rat. AB - This study reports torsion tests and pull-out tests on osseointegrated commercially pure titanium fixtures. The tests were performed in vivo on a total of 26 rats. Three fixtures with a diameter of 2.0 mm were installed bilaterally in the proximal tibia in each animal. The mechanical testing was performed immediately after installation, after 2, 4, 8 and 16 weeks of unloaded healing. The torsional strength started to increase after 4 weeks of unloaded healing and there was a significant increase with time during the initial 16 weeks. The pull out load increased rapidly during the first 4 weeks; thereafter, a moderate increase occurred during the following 12 weeks. A histological evaluation was performed after 0, 4, 8 and 16 weeks. There were significant (P < 0.01) correlations between torque and percentage of bone in contact with the fixture, and between pull-out load and the bone thickness around the fixture (P < 0.001). Estimations of shear stresses and shear moduli in the bone tissue (pull-out test) and at the interface (torque test) indicated that the increase in bone volume around the implant substantially improved the mechanical capacity. PMID- 9212193 TI - Inflammatory reaction dependence on implant localization in rat soft tissue models. AB - This study compares two different implantation models in soft tissue in rat abdominal wall with regard to inflammatory reactions. Titanium rods and discs, penetrating or not penetrating the peritoneal wall respectively, were implanted. After 3, 10 or 30 days the distribution of monocytes/macrophages and cytokines (interleukin-1 and transforming growth factor-beta) in the tissue adjacent to the implants was investigated under immunohistochemistry. The macrophage-specific antibody, ED1, was used for the identification of newly recruited macrophages and the ED2 antibody was used for the mature tissue macrophages. After 10 days the non-penetrating implants had a larger number of cells close to the implant than the penetrating implants. The opposite was seen after 30 days implantation, with a larger number of cells around the penetrating implants. At all time intervals the penetrating implants had a thicker reactive capsule. The cytokines interleukin-1beta and transforming growth factor-beta could be detected in the reactive tissue adjacent to both types of implants, without obvious differences for the two implant situations. The biocompatibility of a material appears to be influenced by the localization of the implant. In addition, it seems to be of importance to extend the follow-up periods further, as we cannot assume that steady state is reached at 30 days implantation. PMID- 9212194 TI - In vitro induction of osteogenic differentiation from non-osteogenic mesenchymal cells. AB - The process of ectopic bone formation suggests that extraskeletal cells are capable of osteogenesis. Alkaline phosphatase (ALP) activity is considered to be an early marker of osteogenic differentiation. This study determined whether cells from the rabbit dermis, striated muscle and extramedullary adipose tissue could undergo osteogenic differentiation in vitro. The cells were cultured with two osteoregulators, recombinant human bone morphogenetic protein 2 (rhBMP2) and dexamethasone. Incubation of extramedullary adipose cells with a combination of rhBMP2 and dexamethasone resulted in an increase in their ALP activity. The results suggest that extramedullary adipocytic cells may undergo osteogenic differentiation. PMID- 9212195 TI - Elastic modulus of the periodontal ligament. AB - This study used a two-dimensional finite element mesh of a lower first premolar to model two different tooth loading systems which measured either the vertical or the horizontal displacements of this particular tooth. The elastic modulus of the periodontal ligament was varied in the finite element model until the horizontal and vertical displacements of the model correlated with the two experimental systems. It was found that an elastic modulus of 50 MPa gave good correlation between the finite element model and the experimental systems. PMID- 9212196 TI - Adsorption of chondroitin-4-sulphate and heparin onto hydroxyapatite--effect of bovine serum albumin. AB - The adsorption of chondroitin-4-sulphate (C4S) and heparin onto hydroxyapatite (HA) has been studied in the absence and presence of bovine serum albumin (BSA). Isotherm data at pH 6.8 have shown that BSA in solution has no effect on C4S adsorption, whereas heparin affinity and adsorption decrease. These data suggest that C4S and BSA bind to different calcium sites on the HA surface. Heparin and BSA may compete for the same calcium sites, or alternatively form heparin-BSA complexes leading to less binding due to steric effects. Evidence of an interaction between heparin and BSA in solution has been shown in this study, there being negligible interaction for C4S. BSA adsorption from solution onto HA decreases with increasing C4S/heparin solution concentration, which may be due to glycosaminoglycan-induced conformational change of BSA from a compact to an extended structure. For the HA precoated with BSA, both C4S and heparin adsorption decrease above a certain solution concentration. A possible explanation is that precoated BSA masks binding sites for the C4S/heparin. The percentage of BSA desorbed from the precoated HA in the presence of C4S and heparin is < 10% and < 30% respectively, indicating that BSA is strongly bound to the HA surface. PMID- 9212197 TI - Effect of chemical structure of hydrogels on the adhesion and phenotypic characteristics of human monocytes such as expression of galectins and other carbohydrate-binding sites. AB - The reactivity of diverse immune aspects to the presence of synthetic polymers represents one of the most important aspects of implantable device biocompatibility. In this report, we show the effect of the chemical structure of a synthetic polymer support on monocyte adhesion and selected phenotypic characteristics in vitro as a model for the initial steps of non-self-recognition of an implant. The extent of monocyte adhesion was significantly influenced by the support chemistry. The highest level of monocyte adhesion was observed on a surface copolymer of 2-hydroxyethyl methacrylate with dimethyl aminoethyl methacrylate relative to results of experiments in which poly(2-hydroxyethyl methacrylate) or the copolymer of 2-hydroxyethyl methacrylate with the sodium salt of methacrylic acid was used. Cell adhesion to the polymers tested and to glass was accompanied by enhanced expression of the carbohydrate-binding sites tested for asialoglycoprotein beta-galactosides such as galectins, beta-N acetylgalactosamine, alpha-mannoside, specific lectin for heparin as well as the lymphokine-macrophage migration inhibitory factor in the monocytes tested. These results suggest the importance of monocyte adhesion to the biomaterial surface for their development into macrophages and further non-self-recognition of the implanted device. PMID- 9212198 TI - Five hundred consecutive carotid endarterectomies: emphasis on vein patch closure. AB - This study reviews 500 consecutive carotid endarterectomies performed in 429 patients in one practice over a 12-year period, emphasis being placed on the technique of vein patch closure and its durability. The records of all such patients were reviewed. Data collected included indication, age, sex, angiogram and duplex scan results, technique of carotid closure, complications within 30 days, and follow-up postoperative duplex scans. The technique emphasized generous exposure, distal arteriotomy, routine shunting, and narrow vein patch angioplasty. The mean patient age was 68 years; 245 (57.1%) were men, and 184 (42.9%) were women. Indications for surgery were transient ischemic attack 256 (51.2%); symptom-free stenosis 144 (28.8%); recovered stroke 60 (12%); and non hemispheric symptoms 40 (8%). The arteriotomy was closed primarily in 71 (14.2%) instances and with a patch in 429 (85.8%). Complications included five (1%) deaths, one (0.2%) stroke, nine (1.8%) transient ischemic attacks, and four (0.8%) wound hematomas. One (0.2%) vein patch rupture occurred. Serial postoperative duplex scans were reviewed in 455 (91%) patients. No significant residual disease was found in any of these patients; three (0.7%) patients were identified with recurrent symptom-free stenoses of >80%; one (0.2%) silent carotid occlusion occurred; and no aneurysms were identified. Classic descriptions of carotid endarterectomy limited the carotid arteriotomy to the bulb area, while contemporary carotid surgery emphasizes wide internal carotid exposure and distal arteriotomy. The authors' experience with vein patch closure confirms the validity of this technique and its low short- and long-term morbidity. PMID- 9212199 TI - Clinical results of Greenfield filter use in patients with cancer. AB - The purpose of this report is to examine the outcomes for patients with an underlying diagnosis of malignancy who have had Greenfield vena caval filters placed for protection from pulmonary embolism, and to identify areas requiring further study. This was a retrospective review of data obtained from the Greenfield filter registry and the University of Michigan Tumor Registry for 166 patients treated at the University of Michigan Medical Center between January 1988 and June 1994. The 84 men and 82 women (mean age 57.8 years) had a mean survival time of 10 (range 1-68) months. This differs significantly from patients in the filter registry who do not have malignancy (P<0.0001). Some 51% experienced recurrence of their malignancy at a mean of 20 months; this timing corresponds to development of new or recurrent thrombembolism and filter placement. Distant metastases were present in 72% of patients at the time of filter placement. In conclusion, as anticipated, filter patients with malignancy have a significantly shorter survival time than those with other concurrent diseases. A temporal association between the progression of the malignancy and the occurrence of thromboembolism is observed in this population and requires further study. Future studies regarding the use of vena caval filters in these patients and the role of diagnostic screening for deep venous thrombosis and occult recurrence of malignancy should focus on efficacy, safety, cost and patient quality of life rather than on survival. PMID- 9212200 TI - Progress in abdominal aortic aneurysm surgery: four decades of experience at a teaching center. AB - The purpose of this study was to examine the changing trends in surgical management of patients with abdominal aortic aneurysms at a tertiary care teaching hospital over the past 40 years, by analysis of demographic data, perioperative variables and outcomes on all patients having abdominal aortic aneurysm surgery between 1955 and 1993. Some 1604 abdominal aortic aneurysms were assessed. The annual rate of abdominal aortic aneurysm surgery increased from 17.6 to 67.8 cases per year. The non-ruptured to ruptured abdominal aortic aneurysm ratio increased from 2.4:1 in the first decade to 3.4:1 in the last 5 years. In non-ruptured abdominal aortic aneurysm repairs, the following variables changed over the four decades: patients age over 80 years increased (2.4% to 8.0%; P<0.04), concomitant lower-limb occlusive disease increased (12.2% to 23.7%; P<0.02), prevalence of smaller aneurysms (4-6 cm) increased (16.0% to 54.2%; P<0.0001); intraoperative hypotension decreased (9.0% to 0.7%; P<0.0001), postoperative hemorrhage decreased (8.2% to 0.0%, P<0.0001), postoperative leg ischemia decreased (5.7% to 1.1%; P<0.02) and postoperative amputation rate decreased (3.2% to 0.0%; P<0.03). There was a significant decrease in perioperative mortality (17.0% to 3.4%; P<0.0001). For ruptured aneurysms, early operation (within 1 h of admission) increased from 8.7% to 55.8% (P<0.0001), prevalence of intraoperative hypotension decreased (50.0% to 23.5%; P<0.001), and major venous injury decreased (18.0% to 5.2%; P<0.05). Mortality, however, did not decrease significantly (54.2% to 44.2%; P=0.32). In conclusion, there was a significant decrease in mortality and morbidity associated with non-ruptured abdominal aortic aneurysm repair over the four decades studied. In addition, older patients with smaller aneurysms and more co-morbid conditions were operated on during this period. Mortality for patients operated on for ruptured abdominal aortic aneurysm repair has not changed significantly. PMID- 9212201 TI - Activated protein C resistance, factor V Leiden and peripheral vascular disease. AB - Activated protein C resistance caused by factor V Leiden is an important thrombophilia disorder which predisposes to venous thromboembolism. Some studies also suggest a role in the pathogenesis of arterial thrombosis and atherosclerosis. The authors have investigated the prevalence of activated protein C resistance and factor V Leiden in a series of 45 patients with peripheral vascular disease. Twelve patients were receiving warfarin. The activated protein C resistance ratios were significantly lower in the group of 33 non-warfarinized patients with peripheral vascular disease (median 2.82 (range 1.36-3.83)) compared with 33 age- and sex-matched controls (median 2.97 range 2.24-4.11); P<0.005; Wilcoxon rank sum). Eight patients (24%) had activated protein C resistance (ratio <2.2). The prevalence of factor V Leiden in patients with peripheral vascular disease was 17.8% (8/45). This is significantly increased compared with the local population and UK published frequency of 3.5% for this genotype. The presence of factor V Leiden did not affect the late outcome of arterial reconstructive surgery in terms of graft patency (P=0.5, Fisher's Exact test). PMID- 9212202 TI - Reduced blood flow accelerates intimal hyperplasia in endarterectomized canine arteries. AB - The purpose of this study was to evaluate a technique that accelerates intimal hyperplasia by reduction of blood flow. Bilateral endarterectomies were performed in both femoral and carotid arteries in six dogs. One week later, all animals underwent banding of an artery distal to the injured region to reduce the blood flow by 50%. The contralateral injured arteries served as controls. At 11 weeks, the specimens were harvested and analyzed. Five of 12 (42%) of the flow restricted arteries and nine of 12 (75%) of the non-flow-restricted arteries were patent at 11 weeks (P<0.05). Marked stenotic intimal hyperplastic lesions developed in the flow-restricted arteries (69% stenosis) as compared with the non flow-restricted arteries (37% stenosis). Mean(s.d.) intimal thickness, intimal areas, and intimal/medial area ratio were 0.52(0.19) mm, 3.17(1.11) mm2, and 1.12(0.33)%, respectively, in the flow-restricted arteries. Their counterparts in the non-flow-restricted arteries were 0.21(0.09) mm, 1.70(1.09) mm2, and 0.58(0.14)%, respectively (P<0.05). Extracellular matrix comprised 48% of total intimal volumes in the flow-restricted arteries. Cell proliferation and occluded arteries were also characterized. These data demonstrate that reduction of blood flow significantly accelerated intimal hyperplasia and occlusion rates in endarterectomized arteries. Advanced intimal hyperplastic lesions (>50% stenosis) possess a high extracellular matrix content. This new animal model is a reliable generator of advanced stenotic lesions in a relatively short time period and can be used to study biologic mechanisms of stenosis and evaluate therapeutic interventions. PMID- 9212203 TI - An arginine analog inhibits responses of both the endothelium and smooth muscle of canine in situ vein grafts. AB - Increased shear, pressure, and oxygen tension in vein grafts may alter production of endothelium-derived vasoactive and anti-mitogenic factors such as nitric oxide which subsequently affect development of neointimal hyperplasia. This study was designed to determine whether or not nitric oxide mediates endothelium-dependent responses in femoral in situ vein grafts. Non-reversed, canine femoral vein grafts were placed bilaterally to bypass a ligated segment of the femoral artery in dogs. After 6 weeks, the grafts were removed, cut into rings, and suspended in organ chambers for measurement of isometric force. In some rings the endothelium was removed deliberately. In the presence of indomethacin, the synthetic analog of L-arginine, L-N(G)-monomethyl-arginine (10(-4) M; L-NMMA) did not cause a significant change in baseline tension of rings with endothelium. L-NMMA reduced only contractions of rings with endothelium to the alpha-adrenergic agonist UK 14,304. The analog also reduced maximal relaxations to the calcium-ionophore, A23187 in rings with endothelium. In addition, L-NMMA reduced relaxations of rings without endothelium to adenosine diphosphate by 35%. Positive immunostaining for nitric oxide synthase was present in both the myointima and media of histological sections of grafts. In conclusion, these results suggest that in situ vein grafts exhibit two unique properties which are unlike unoperated arteries or veins: (i) alpha2-adrenergic receptors may be coupled to the release of contractile endothelium-derived factors associated with production of nitric oxide: and (ii) nitric oxide may be released by the smooth muscle in response to purinergic stimulation. The presence of nitric oxide synthase throughout the wall of the graft may result in production of nitric oxide in response to adenosine diphosphate released by platelets and to circulating catecholamines. PMID- 9212204 TI - Inhibitory effect of type 1 collagen gel containing alpha-elastin on proliferation and migration of vascular smooth muscle and endothelial cells. AB - The purpose of this study was to investigate in vitro the potential effect of type 1 collagen gel containing alpha-elastin on the proliferation of vascular smooth muscle cells and vascular endothelial cells, and on smooth muscle cell migration. Vascular smooth muscle cell and endothelial cell were cultured in 12 well plates precoated with collagen gels and alpha-elastin. Cell proliferation rates were measured by monitoring [3H]-thymidine incorporation. After 2, 3 or 4 days of culture, the proliferation rate of both smooth muscle cells and endothelial cells was significantly decreased on collagen gel containing 10 mg/ml alpha-elastin compared with collagen gel only as control. Smooth muscle cell proliferation on collagen gel containing alpha-elastin on the 4th day of culture was decreased dose-dependently, e.g. 1 mg/ml of alpha-elastin (74.8(2.3)% of control, P=n.s.); 5 mg/ml (56.7(2.1)%; P<0.05); 10 mg/ml (30.3(3.1)%; P<0.005). In the case of cultured endothelial cells, however, [3H]-thymidine incorporation was not decreased significantly in the presence of 5 mg/ml alpha-elastin (83.1(7.9)%, P=n.s.). After stimulation by platelet-derived growth factor, the smooth muscle cell migration rate on collagen gel containing alpha-elastin (5 mg/ml) was decreased over time. The area of migration on the 6th day of culture was also significantly decreased dose-dependently in the presence of alpha elastin, e.g. 1 mg/ml (72.6(3.4)% of control, P<0.05), 5 mg/ml (56.9%(1.5)%; P<0.05); 10 mg/ml (37.3(2.7)%; P<0.0005). In conclusion, alpha-elastin inhibited the proliferation and migration of smooth muscle cell in a dose-dependent manner on collagen gel culture, however, at high concentrations of alpha-elastin (10 mg/ml), the endothelial cell proliferation rate was also inhibited. At 5 mg/ml, alpha-elastin significantly inhibited smooth muscle cell proliferation and migration but did not significantly inhibit endothelial cell proliferation. Incorporation of collagen gel containing alpha-elastin into the structure of arterial prosthesis offers the possibility of inhibiting smooth muscle cell hyperplasia without significant effect on endothelial cell formation. PMID- 9212205 TI - Effects of albumin coating of knitted Dacron grafts on transinterstitial blood loss and tissue ingrowth and incorporation. AB - Transinterstitial blood loss at implantation and the degree of graft incorporation and inner capsule thickening was compared in serial explants of albumin-coated Dacron versus blood preclotted Dacron grafts in the canine thoracoabdominal aortic position (8 mm internal diameter x 30 cm length). The coated grafts were Bard DeBakey Vasculour II knitted Dacron prostheses impregnated with carbodiimide-cross-linked human albumin. Control grafts were otherwise identical and preclotted with the recipients' whole blood before heparinization during surgery. Transinterstitial blood loss after establishing flow was measured by weighing sponges wrapped around the grafts. Albumin pretreatment resulted in significantly less median blood loss (5.1 g versus 11 g, P=0.04; Mann-Whitney rank sum test). Grafts were explanted at 1 week, 4 weeks, 10 weeks, and 20 weeks. Patency was 100% in both groups. Graft incorporation at explantation was graded by the surgeon as: 1 = none, 2 = minimal, 3 = moderate, or 4 = extensive. No significant differences were noted at any time period. Inner capsule thickness measurements were made every 2.5 mm along the length of all explants. Grafts explanted at 1 week displayed no inner capsules. By 20 weeks, median inner capsule thickness was significantly less in albumin-coated grafts (190 microm versus 235 microm; P<0.0001). These inner capsules in both groups formed as islands, containing abundant myofibroblasts and collagen, covered by endothelial cells and surrounded by residual fibrin coagula. In conclusion, albumin-coated knitted Dacron grafts displayed less transinterstitial blood loss at implantation, and qualitatively similar incorporation, but significantly thinner inner capsules at 20 weeks. PMID- 9212206 TI - Surgical aspects of fusiform and saccular extracranial carotid artery aneurysms. AB - The purpose of this paper is to analyse surgical aspects of aneurysms of the distal extracranial internal carotid artery. Nine cases of extracranial carotid artery aneurysm are reported. Five were fusiform, located at the carotid bifurcation, and four were saccular, confined to the internal carotid artery. An end-to-end plication technique and Dacron patch angioplasty were employed for all fusiform aneurysms. In three saccular lesions, resection and 4-mm polytetrafluoroethylene (PTFE) graft interposition were carried out. In one case with a high lesion, ligation of the carotid artery was performed. Ligation resulted in severe postoperative stroke and fatal outcome. One patient with a saccular lesion developed a transient ischaemic attack after the operation. In other patients no central neurological deficit was produced by the surgery itself. Transient cranial nerve damage occurred in four patients (two hypoglossal nerve: two superior laryngeal nerve). As demonstrated by these cases, synthetic material may be used in restoration of the carotid artery. It is concluded that, according to type, location of the aneurysm and adequacy of contralateral cerebral blood flow, selective management is necessary. PMID- 9212207 TI - Carotid arterial trauma: assessment with the Glasgow Coma Scale (GCS) as a guide to surgical management. AB - Management of carotid arterial injuries associated with focal neurological deficit or altered state of consciousness (SCON) remains unresolved. Experience with these injuries in one particular hospital was reviewed and the Glasgow Coma Scale (GCS) utilized to assist with clinical stratification of these patients. A literature review was also conducted to better define indications for repair or ligation of carotid injuries. From 1978 to 1990, 34 patients with carotid arterial injuries were reviewed with reference to the GCS, focal deficit, hypotension, anatomic site and mechanism of injury. The literature from 1952 to 1993 was surveyed for carotid artery injuries (1316 patients). Outcome of treatment with or without repair was compared with pre-operative neurologic status. Thirty-four patients with injuries of the common (24) or internal (10) carotid arteries were managed with repair (68%), ligation (24%) or observation (9%). The SCON was normal in 18 patients; 16 patients (88%) underwent repair and all remained normal. All patients with GCS 9-14 regained a normal SCON after surgical repair, while 10 patients with GCS < 8 had repair (5), ligation (3), and non-operative management (2); five returned to normal, four died and one remained comatose. However, outcomes correlated poorly with management. Of 1316 patients cited in the surgical literature, patients with no deficit and patients with pre operative deficits did significantly better after repair as compared with ligation (P<0.001). In comatose patients, management did not affect outcome. It is concluded that carotid arterial injuries should be repaired in patients with normal neurologic evaluation, focal pre-operative neurologic deficits and in patients with GCS > 9. Comatose patients with GCS < 8 do poorly regardless of management. The GCS provides an objective for stratification of patients with altered SCON who benefit from repair of carotid arterial injuries. PMID- 9212208 TI - Ultrasound enhancement of rabbit femoral artery thrombolysis. AB - Experiments were conducted to evaluate the potential of low-intensity, externally applied ultrasound to accelerate arterial thrombolysis in an animal model and to characterize potential effects of ultrasound exposure on vessel wall morphology. The femoral arteries of 32 rabbits were exposed, a flowprobe was positioned around the vessel, and a stenosis produced with two circumferential silk sutures to reduce flow by 50%. Thrombosis was achieved by injecting thrombin through the cannulated superficial epigastric branch into a 1-cm segment of femoral artery which was isolated for 20 min. Streptokinase was administered intravenously as a 15,000 U/kg bolus followed by an infusion of 15,000 U/kg per h. Ultrasound (1 MHz, 2 W/cm2) was delivered to the thrombosed vessel during streptokinase administration in 17 animals, and 15 control animals received sham ultrasound only. Thrombolysis occurred in nine of 17 (53%) animals receiving both streptokinase and ultrasound, and this was significantly greater than the rate in animals receiving streptokinase alone (2/15, 13%; P=0.025). Ultrasound caused a mean temperature elevation of 4 degrees C in exposed tissues. Light and electron microscopy demonstrated increased platelet accumulation on thrombi in ultrasound treated vessels compared with controls. Endothelial cell vacuolation was seen by electron microscopy in ultrasound-exposed vessels. The results indicate that externally applied, low-intensity ultrasound can significantly enhance thrombolysis in a rabbit arterial model. Possible adverse effects are minor and include platelet accumulation, temperature elevation and minor endothelial changes. Externally applied ultrasound has potential value as an adjunct to thrombolytic therapy. PMID- 9212209 TI - Implication of myocardial lactate metabolism during CABG (Ando et al.) PMID- 9212210 TI - Implication of myocardial lactate metabolism during coronary artery bypass grafting. AB - Sixty-six consecutive patients with coronary artery disease were analysed in terms of myocardial lactate extraction during cardiac surgery. Sixteen patients had left main coronary heart disease and 50 were without such disease. Mean (s.d.) lactate extraction during empty beating in patients with and without left main coronary tract disease was -29.8(67)% and 12.0(15.3)%, respectively (P<0.001). No significant differences in lactate extraction were recognized during 15 min of reperfusion. Mean (s.d.) preoperative values of haemoglobin were 11.2(1.0) g/dl and these fell to 6.5(0.9) g/dl in an empty beating state during cardiopulmonary bypass (P<0.001). There was no significant difference between the two groups in preoperative and postoperative left ventricular stroke work index. An empty beating state before aortic clamping could induce unexpected ischaemia in the heart with left main coronary tract disease. A short duration of this ischaemic state does not influence functional recovery; however, exposing the left main coronary artery diseased heart to such a condition for long periods would be dangerous. PMID- 9212211 TI - Admission to an intensive care unit after transvenous implantable cardioverter defibrillator implantation: analysis of risk factors. AB - A retrospective review was undertaken of 90 patients admitted to the cardiothoracic intensive care unit and who comprised 47% of all transvenous implantable cardioverter defibrillator operations performed between March 1991 and August 1995. The review aimed to evaluate the necessity for routine postoperative intensive care unit (ICU) admission after implantable cardioverter defibrillator operation. Pre-, intra- and postoperative data were analysed. Eight of 90 patients (9%) subsequently required care unique to the ICU. None of the variables examined, including the Acute Physiology Score (APACHE II system), was helpful in identifying patients who required unique ICU services. Patients undergoing transvenous implantable cardioverter defibrillator surgery are identified as a low risk group requiring in over 90% of cases monitoring services rather than active therapy in an ICU. As no reliable predictors seem to exist to identify the necessity for postoperative ICU admission, cardioverter defibrillator implantations should only be performed in hospitals where adequate facilities are readily available. PMID- 9212212 TI - The stentless Bravo 300 aortic porcine xenograft: supra-annular versus annular implantation. AB - The aim of the study was to compare the supra-annular and intra-annular implantation techniques by evaluating the differences in early haemodynamic outcome (gradients, effective orifice area, regurgitation). Since August 1991, 200 stentless Bravo model 300 valves have been implanted. Patients were divided into three groups of consecutive cases: group 1 (n = 50) represents exclusively intra-annular implantation; group 2 (n = 50) is a transitional period: and group 3 (n = 100) comprises only patients with supra-annular implantation. Significant differences were found (P<0.001) in low postoperative gradients (mean < or = 10 mmHg): 24% in group 1, 42% in group 2, and 95% in group 3. Comparing groups 1 and 3, gradients were lower and effective orifice area was augmented in all valve sizes in group 3. Trivial central regurgitation was present in groups 1-3 (6%, 12% and 0% respectively). Peripheral regurgitation was trivial in 6%, 8% and 0% and mild to moderate in 4%, 2% and 0% (P<0.001). PMID- 9212213 TI - Surgical treatment of cardiac myxomas: a 20-year follow-up. AB - Clinical experience in the diagnosis and management of 50 cases of cardiac myxoma seen over a 20-year period from 1974 to 1994 has been reviewed. There were 17 men and 33 women of mean age 55.2 (range 16-81) years. Echocardiography confirmed the diagnosis in all patients. The location of myxomas was as follows: left atrial alone in 42 patients, right atrial alone in three, right ventricular alone in one, left atrial+right atrial in two, left atrial+right atrial+right ventricular in one, and left atrial+left ventricular in one. Nineteen patients were operated on within 48 hours of the diagnosis. All tumours were successfully removed with the aid of cardiopulmonary bypass. The hospital mortality rate was 10%. Excision of the tumour resulted in marked symptomatic improvement. There was one late death. The current survivors are symptom-free at a mean follow-up of 76.4 (range 1-241) months. Echocardiographic studies were performed in all survivors and no recurrences have been observed. It is concluded that excision of cardiac myxomas is curative and long-term survival is excellent. Radical tumour excision may prevent recurrences. PMID- 9212214 TI - Stentless valve replacement in the small aortic root. AB - Despite the variety of different artificial heart valves available, no ideal prosthesis for the small aortic root has yet been identified. The aim of this study was to evaluate the haemodynamic performance and clinical outcome after stentless aortic valve replacement. A total of 70 patients with a small aortic root underwent Toronto (n = 61) or Freestyle (n = 9) stentless aortic valve replacement. All but three patients had aortic stenosis. Mean (s.d.) age at operation was 71.2(7.9) years. The mean annular diameter was 21.4(1.2) mm. Using controlled oversizing adjusting valve size to the sinotubular junction diameter, a 23-mm prosthesis was implanted in 23 patients and a 25-mm prosthesis in 47 patients. The maximum pressure gradient was 19.1(6.8) mmHg and effective valve orifice area was 1.47(0.27) cm2. At discharge and at follow-up, all patients were in New York Heart Association class I or II. At follow-up there was a significant reduction in pressure gradients, an increase in effective valve orifice areas, and decrease of pre-existing left ventricular hypertrophy. In conclusion, with controlled oversizing a gain in prosthesis size of 2 to 4 mm can be achieved. Implantation of oversized stentless valves leads to improved haemodynamics and to left ventricular remodelling in patients with a small aortic root. PMID- 9212215 TI - The isolated post ischaemic rat heart is more vulnerable to protamine sulphate than the non-ischaemic heart. AB - Protamine sulphate is currently used for the reversal of heparin anticoagulation but is known to cause direct myocardial depression. The purpose of this study was to compare the effects of protamine sulphate on the isolated heart with and without cardioplegic ischaemia. Isolated rat hearts (Langendorff preparation) were electrically paced at 300 beats/min and perfused with Krebs-Henseleit solution. Five groups were tested: (1) control: no ischaemia, no protamine; (2) no ischaemia, protamine; (3) no ischaemia, protamine (time-matched control to groups 4 and 5); (4) control: ischaemia, no protamine; and (5) ischaemia, protamine. Protamine sulphate was infused for 15 min at 10 microg/ml. In groups 4 and 5, cardioplegic ischemia was maintained for 30 min at 30 degrees C before protamine exposure. Protamine decreased myocardial performance in a time- and dose-dependent manner. Protamine depressed mean (s.d.) myocardial left ventricular pressure in both non-ischaemic hearts (groups 2 and 3, to 49(4)% and 50(4)% from baseline, respectively) and post ischaemic hearts (group 5, to 28(8%). Mean (s.d.) left ventricular-developed pressure only partially recovered after protamine in post-ischaemic hearts (to 55(13)% of baseline) compared with full recovery of the non-ischaemic group. Protamine depressed coronary flow to 70(5)% and 74(8)% in non-ischaemic hearts (groups 2 and 3, respectively) and to 58(7)% in group 5. Coronary flow recovered completely at the end of the experiments in all protamine-treated groups. In conclusion, isolated rat hearts subjected to cardioplegic ischaemia are more vulnerable to protamine than are non ischaemic hearts. PMID- 9212216 TI - The urokinase-type plasminogen activator system in cancer metastasis: a review. AB - The urokinase-type plasminogen activator (u-PA) system consists of the serine proteinases plasmin and u-PA; the serpin inhibitors alpha2-anti-plasmin, PAI-1 and PAI-2; and the u-PA receptor (u-PAR). Two lines of evidence have strongly suggested an important and apparently causal role for the u-PA system in cancer metastasis: results from experimental model systems with animal tumor metastasis and the finding that high levels of u-PA, PAI-1 and u-PAR in many tumor types predict poor patient prognosis. We discuss here recent observations related to the molecular and cellular mechanisms underlying this role of the u-PA system. Many findings suggest that the system does not support tumor metastasis by the unrestricted enzyme activity of u-PA and plasmin. Rather, pericellular molecular and functional interactions between u-PA, u-PAR, PAI-1, extracellular matrix proteins, integrins, endocytosis receptors and growth factors appear to allow temporal and spatial re-organizations of the system during cell migration and a selective degradation of extracellular matrix proteins during invasion. Differential expression of components of the system by cancer and non-cancer cells, regulated by paracrine mechanisms, appear to determine the involvement of the system in cancer cell-directed tissue remodeling. A detailed knowledge of these processes is necessary for utilization of the therapeutic potential of interfering with the action of the system in cancers. PMID- 9212217 TI - Prevalence of p53 mutations and protein expression in esophageal cancers in southern Thailand. AB - To investigate p53 alterations in esophageal squamous-cell carcinomas of patients in the high-risk area of southern Thailand, 72 paraffin-embedded samples were analyzed immunohistochemically for p53 protein expression and 16 frozen samples for p53 mutational status. Forty-two of the 72 tumors (58.3%) showed p53 protein accumulation in the nuclei of tumor cells. Expression of p53 in tumors was not significantly correlated with gender, histological grading, depth of invasion, node involvement, smoking or alcohol consumption. Analysis of the p53 gene in a sub-set of 16 tumors showed mis-sense mutations in 7 out of 11 p53-positive and 1 out of 5 p53-negative tumors. The p53 mutational spectrum was 50% transitions (3 C-to-T and 1 G-to-A, all occurring at CpG dinucleotide sites) and 50% transversions (one each, C-to-G, G-to-T, T-to-G, and T-to-A). Our findings support the hypothesis that alterations of p53 are involved in the carcinogenesis of most squamous-cell carcinomas of the esophagus, irrespective of the population and the factors responsible for carcinogenesis. The mutation profile of the p53 gene might indicate etiologic contributions of different mutagen exposures in patients from high-risk areas of southern Thailand. PMID- 9212218 TI - Loss of heterozygosity on chromosome 9 and p16 (MTS1, CDKN2) gene mutations in esophageal cancers. AB - Loss of heterozygosity on chromosome 9 has been reported in a variety of human cancers. The cyclin-dependent kinase inhibitor p16 gene, mapped on chromosome 9p21, is presumed to be the tumor-suppressor gene localized in this chromosome. The aim of our study was to determine, in 26 Barrett's adenocarcinomas and 20 squamous-cell carcinomas of the esophagus, the prevalence of loss of heterozygosity on chromosome 9 by typing of microsatellite loci and mutation of p16 by direct sequencing of exons 1 and 2. Allelic losses were found in 69% of adenocarcinomas, but only a microdeletion in exon 1 of p16 occurred in 1 tumor. Among squamous-cell carcinomas, 65% had allelic losses and 5 tumors were mutated on the p16 gene (1 deletion, 3 nucleotide substitutions and 1 insertion). The relatively low rate of p16 mutation observed here coupled with the high frequency of loss of heterozygosity on chromosome 9 suggests that one or several tumor suppressor gene(s) distinct from p16 may be the target(s) of allelic deletion in most esophageal cancers or that p16 is inactivated in another way. PMID- 9212219 TI - Detection of 14q32.33 translocation and t(11;14) in interphase nuclei of chronic B-cell leukemia/lymphomas by in situ hybridization. AB - Abnormalities of chromosome 14 involving band q32.33 are among the most commonly observed cytogenetic alterations in B-cell malignancies. To assess the incidence and pathogenetic implications of 14q32.33 translocation in chronic B-cell leukemia/lymphomas, we performed fluorescence in situ hybridization (FISH) analysis with variable region (V(H)) and gamma constant region (Cgamma) gene probes in 37 patients with these disorders. Chromosome 14q32.33 translocation was detected in 2 of 18 patients with chronic lymphocytic leukemia (CLL), 1 of 2 with CLL of mixed cell types (CLL/PL), 1 of 2 with pro-lymphocytic leukemia (PLL), 5 of 6 with leukemic mantle-cell lymphoma (MCL), 2 of 7 with splenic B-cell leukemia/lymphoma of possible marginal zone origin (SBLL) and 2 with leukemic follicular lymphoma (FL). To further characterize 14q32.33 translocations in these patients, we developed a new procedure using double-color FISH with PRAD1, BCL2, V(H) and Cgamma gene probes. Chromosome t(11;14) was detected in 1 patient with CLL/PL, 1 with PLL and 5 with MCL. Chromosome t(14;18) was detected in 2 patients with FL. In a PLL patient with t(11;14), the cosmid CPP29 containing the PRAD1 gene and its 5'-flanking region split and co-localized with both Cgamma and V(H) gene probes, thus spanning the breakpoint. In CLL and SBLL patients, donor chromosomes were other than chromosomes 2, 11, 18 and 19, suggesting the involvement of a novel oncogene(s) in the pathogenesis of these diseases. Interphase FISH rapidly detected 14q32.33 translocation, t(11;14) and t(14;18) in B-cell malignancies with low mitotic activity at the single-cell level, facilitating the correlation of the molecular features of these translocations with clinical characteristics. PMID- 9212220 TI - Recovery of leukocyte function after super-high-dose chemotherapy with peripheral blood stem cell transplantation in testicular cancer patients. AB - Leukocyte functions (superoxide production and phagocytosis) were evaluated in patients with testicular cancer who had undergone super-high-dose chemotherapy (SHDT) with peripheral blood stem cell transplantation (PBSCT). In 5 patients with advanced or relapsed testicular cancer who received 8 cycles of SHDT with PBSCT, we measured superoxide production activity by phorbol myristate acetate (PMA) stimulation, as well as phagocytic activity using latex particles before and after PBSCT. The median time to reach a leukocyte count of > or = 1,000/microl was 9 days after PBSCT. Superoxide production activity was significantly decreased on day 7 compared with the pre-chemotherapy level. However, it recovered by day 12 and sustained such a level thereafter. Phagocytic activity was within the normal range on day 7 but declined slightly on days 12 and 21 compared with the pre-chemotherapy level. Our results indicate that leukocyte function and leukocyte count recover promptly following PBSCT and that PBSCT is a safe and convenient adjunctive therapy after SHDT for patients with advanced testicular cancer. PMID- 9212221 TI - Expression of type IV collagen alpha1(IV)-alpha6(IV) polypeptides in normal and developing human kidney and in renal cell carcinomas and oncocytomas. AB - Type IV collagen trimer is a major component of basement membranes (BMs). It is composed of polypeptides named alpha1(IV)-alpha6(IV) chains. Chains alpha1,2(IV) are widely expressed in BMs while alpha3(IV)-alpha6(IV) are more restricted in human tissues. We have now studied by immunohistochemical means the distribution of collagen IV chains in fetal and adult human kidney, in oncocytomas, in renal cell carcinomas (RCCs) and their metastases and in experimental xenografts of human tumors. alpha1,2(IV) chains were found in all BMs of fetal and adult kidney as well as of renal tumors, while alpha3(IV)-alpha6(IV) chains were found in BMs of distal segments of developing and mature tubules. alpha3(IV)-alpha5(IV) chains were seen also in BMs of developing fetal glomeruli after the capillary loop stage. Most of the RCCs and their metastases showed occasional expression of alpha3(IV)-alpha6(IV) with papillary variants showing only expression of alpha5(IV) chain. There was a distinct expression of alpha3(IV)-alpha5(IV) chains in BMs of 3 oncocytomas. In 2 of them a variable expression of the alpha6(IV) chain was seen. In 3 of 4 xenografts, immunoreactivity for human-specific monoclonal antibody (MAb) for alpha1,2(IV) was seen in the BM-like structures. No alpha3-alpha6(IV) was seen in any of the xenografts, while polyclonal antiserum for type IV collagen presented immunoreactivity in BMs of all xenografts. Our results show that oncocytomas and most of the RCCs express scarce variants of type IV collagen containing alpha3(IV)-alpha6(IV) chains. In experimental xenograft tumors, both implanted RCC cells and host stromal cells have a capacity to produce type IV collagen. PMID- 9212222 TI - Heterosexual transmission of hepatitis C virus among married couples in southwestern Japan. AB - The heterosexual transmission of hepatitis C virus (HCV) remains controversial, and data from general populations are scanty. In this cross-sectional study, we assessed the seroprevalence of antibodies to hepatitis C virus (anti-HCV) and the presence and genotype of HCV-RNA among 109 married couples within an endemic, community-based Japanese population. Overall, 25% of the husbands and 32% of the wives had anti-HCV. Spouses with anti-HCV-positive partners were around 2 times more likely to have anti-HCV than spouses with anti-HCV-negative partners (p = 0.01). Of 6 couples in which both spouses had HCV-RNA, however, 3 presented discordant HCV genotypes (type 1b vs. 2b). The couples' anti-HCV concordance status was not significantly influenced by the presence or absence of HCV-RNA among anti-HCV-positive partners (odds ratio [OR]: 0.8 for wives, 0.6 for husbands), nor by the length of marriage, the number of pregnancies or the use of contraceptives. No significant associations with anti-HCV were observed for serum markers of sexually transmitted agents, including human T-lymphotropic virus (OR = 1.1, 95% confidence interval [CI] 0.5-2.3), Treponema pallidum (OR = 0.7; CI 0.1-6.1) and hepatitis B virus (OR = 1.6; CI 0.9-3.0). Our results suggest that the clustering of HCV infection among specific couples within this endemic population may not be attributable to heterosexual transmission. Follow-up studies are necessary to determine the risk of heterosexual transmission of HCV in endemic areas. PMID- 9212223 TI - Food groups and risk of colorectal cancer in Italy. AB - The proportion of colorectal cancer attributed to dietary habits is high, but several inconsistencies remain, especially with respect to the influence of some food groups. To further elucidate the role of dietary habits, 1,225 subjects with cancer of the colon, 728 with cancer of the rectum and 4,154 controls, hospitalized with acute non-neoplastic diseases, were interviewed between 1992 and 1996 in 6 different Italian areas. The validated food-frequency questionnaire included 79 questions on food items and recipes, categorised into 16 food groups. After allowance for non-dietary confounding factors and total energy intake, significant trends of increasing risk of colorectal cancer with increasing intake emerged for bread and cereal dishes (odds ratio [OR] in highest vs. lowest quintile = 1.7), potatoes (OR = 1.2), cakes and desserts (OR = 1.1), and refined sugar (OR = 1.4). Intakes of fish (OR = 0.7), raw and cooked vegetables (OR = 0.6 for both) and fruit other than citrus fruit (OR = 0.7) showed a negative association with risk. Consumption of eggs and meat (white, red or processed meats) seemed uninfluential. Most findings were similar for colon and rectum, but some negative associations (i.e., coffee and tea, and fish) appeared stronger for colon cancer. Our findings lead us to reconsider the role of starchy foods and refined sugar in light of recent knowledge on the digestive physiology of carbohydrates and the insulin/colon cancer hypothesis. The beneficial role of most vegetables is confirmed, with more than 20% reduction in risk of colorectal cancer from the addition of one daily serving. PMID- 9212224 TI - A case-control study of self-reported exposures to pesticides and pancreas cancer in southeastern Michigan. AB - A case-control study of pancreas cancer in residents, aged 30-79 years, of 18 counties in southeastern Michigan was conducted to investigate the risks of exposure to DDT and related materials in the general population. Sixty-six people with cytologically diagnosed pancreas cancer were identified using 7 participating hospitals in metropolitan Detroit and Ann Arbor. One hundred and thirty-one controls were frequency-matched to the cases on age, sex, ethicity and county of residence by random-digit dialing. All study participants were administered a questionnaire to assess life-time exposure to pesticides from both environmental and occupational sources, family history of cancer, past medical history, smoking history and demographic information. A statistically significant increased risk was found for self-reported exposure to ethylan (1,1-dichloro-2,2 bis(4-methoxyphenyl) ethane). Increased odds ratios were observed for self reported exposures to chloropropylate and DDT, as well as for the summary group of organochlorine pesticides which included all of these materials, though these associations were not significant. PMID- 9212225 TI - Expression of HHV-8 latency-associated T0.7 RNA in spindle cells and endothelial cells of AIDS-associated, classical and African Kaposi's sarcoma. AB - Analysis by polymerase chain reaction (PCR) and serological studies have demonstrated a close association between the novel human herpes virus, Kaposi's sarcoma-associated herpes virus (KSHV) or human herpes virus-8 (HHV-8) and the development of Kaposi's sarcoma (KS). To clarify the role of HHV-8 in KS pathogenesis, we investigated at the cellular level by in situ hybridization the expression of a recently described 0.7-kb HHV-8-encoded mRNA (T0.7 mRNA) in KS tissues of different epidemiological origin (AIDS-KS, African endemic KS and classical KS). The T0.7 mRNA likely encodes a small membrane protein, supposedly expressed in latently HHV-8-infected cells. Indeed, we detected T0.7 mRNA in virtually all cells of the cell line BCBL-1 established from a body cavity-based lymphoma (BCBL) and latently infected with HHV-8. In all KS biopsies examined, independent of their epidemiological type, the late-stage (nodular) KS tissues showed a high level of T0.7 mRNA expression in typical KS spindle cells but also in endothelial cells lining blood vessels, indicating latent HHV-8 infection of these cells. The presence of T0.7-expressing cells was restricted to KS tumor tissue and therefore appears to indicate an important role of latent HHV-8 infection in KS pathogenesis. PMID- 9212226 TI - Molecular epidemiology of Epstein-Barr virus (EBV) in EBV-related malignancies. AB - The prevalent strain of Epstein-Barr virus (EBV) in EBV-related malignancies and in healthy adults in Southern Japan was examined by means of polymerase chain reaction (PCR) and/or restriction fragment length polymorphism (RFLP) analysis. In EBV-related gastric cancers, 51/73 cases were subtype A, 4 were subtype B and the EBNA-2 region was not amplified in 18 cases. Sixty-three were wild-type F, and only one was variant "f". Sixty-one cases had type C and 2 type D. EBNA-2 subtype A was found in 10/12 EBV-related T/NK-cell lymphomas, and 11 samples harbored the wild-type F. Neither subtype B nor the "f" variant was detected. Type C EBV was found in 8 cases and type D in 3 specimens. Two Japanese nasopharyngeal carcinomas (NPC) harbored subtype A with wild-type F and type C. Throat washings from healthy adults harbored wild-type F virus in 60/153 cases, and 25 of these samples were EBNA-2 subtype A. Type C viruses were detected in 92% of cases and type D in 7.4%. Therefore, the prevalent strain in EBV-related malignancies in Southern Japan was the same as in the healthy population in this geographical region. PMID- 9212227 TI - Pleural mesothelioma mimics the integrin profile of activated, sessile rather than detached mesothelial cells. AB - Mesothelial cells (MC) form a polarized monolayer lining serosal cavities. During serositis, the MC lining undergoes hyperplasia, and MC are shed into effusions. During these processes, contact with basement membrane and, ultimately, neighboring cells is at least temporarily lost, suggesting regulated alterations in cell/matrix and cell/cell adhesion. Such interactions are primarily mediated by integrins. Malignant mesothelioma has a growth pattern characterized by lateral, limited invasive but contiguous spread. During serositis, activated MC, both sessile and detached, expressed an extended spectrum of beta1, beta3 and beta4 integrins compared with resting MC, as shown by immunohistology. Malignant mesothelioma had an integrin repertoire and a subcellular distribution resembling that of activated sessile rather than floating MC. In vitro, MC exposed a more comprehensive pattern of integrins than that of the newly established mesothelioma cell lines ME-HD-1 and ME-HD-2, as shown by flow cytometry. MC consistently adhered better than mesothelioma cells to laminin, tenascin, fibronectin and collagen type IV. Adhesion of MC and mesothelioma cells to these matrix proteins was, at least in part, mediated via beta1 integrins. The different integrin profiles and adhesion properties of cultured MC and mesothelioma cells may reflect a limited functional differentiation of the latter. PMID- 9212228 TI - Comparison of MUC-1 mucin expression in epithelial and non-epithelial cancer cell lines and demonstration of a new short variant form (MUC-1/Z). AB - Mucins, including MUC-1, are generally considered to be products of epithelial tissues and of their tumors. To examine the possible expression of MUC-1 in other cell types, a panel of human epithelial and non-epithelial tumor cell lines was studied by reverse transcriptase polymerase chain reaction (RT-PCR), Northern blot analysis, immunocytology and radioimmunoprecipitation. Using the highly sensitive RT-PCR method, products corresponding to the non-repetitive 5' and 3' MUC-1 sequences were detected in all the cell lines examined. Amplified products lacking the tandem repeat region of MUC-1, including a new short form (designated MUC-1/Z) different from the previously reported MUC-1/Y protein, were also detected in most cell lines tested. Northern blot analysis, using a probe to the variable number tandem repeat (VNTR) region, confirmed the presence of MUC-1 mRNA in the astrocytoma, melanoma and neuroblastoma cell lines studied. MUC-1 protein was detected by immunocytology in these cell lines using monoclonal antibody (MAb) 139H2. Immunoprecipitation analysis with [3H]-glucosamine-labeled cell lysates and MAb 139H2 or an antibody to the cytoplasmic domain, CT-1, detected MUC-1 protein in 2 epithelial cell lines, an astrocytoma cell line (SK-MG-4) but not in the melanoma and neuroblastoma cell lines studied. Northern blot analysis using a probe to the 3' end of MUC-1 mRNA, confirmed the presence of MUC-1 mucin and also identified short products corresponding to the size of the short variant forms. Protein products corresponding to the MUC-1/Y and MUC-1/Z variant forms were not observed using either [3H]-glucosamine-labeled OVCAR-3 cells or [3H] amino acid-labeled MCF-7 cells and either CT-1 antibody or MAb 232A1, detecting an epitope to the C-terminal region. Thus, depending on the sensitivity of the assay used, varying amounts of MUC-1 mRNA and protein could be detected in non epithelial tumor cell lines. Although the amounts of MUC-1 in these cell lines are much lower than in carcinomas, it is possible that MUC-1 mucin serves a similar function in non-epithelial as in epithelial cells. The possible role of MUC-1/Y and MUC-1/Z variant forms in these cell lines is not understood. PMID- 9212229 TI - Chemopreventive efficacy of anethole trithione, N-acetyl-L-cysteine, miconazole and phenethylisothiocyanate in the DMBA-induced rat mammary cancer model. AB - The chemopreventive efficacy of N-acetyl-L-cysteine (NAC), anethole trithione, miconazole and phenethylisothiocyanate (PEITC), each of which would be expected to alter carcinogen metabolism, was examined in the dimethylbenzanthracene (DMBA) mammary carcinogenesis model. In this protocol, animals were exposed to non-toxic doses of the chemopreventives in the diet beginning 7 days prior to DMBA administration and then continuously throughout the duration of the assay (100 days post carcinogen). Miconazole, an antifungal agent with relatively broad inhibitory activity toward a variety of cytochromes P450, increased mammary tumor latency, decreased tumor incidence at the highest dose and decreased tumor multiplicity up to 60%. Anethole trithione, a substituted dithiolthione and an analog of the relatively broad-spectrum chemopreventive oltipraz, was administered in the diet and significantly inhibited mammary cancer multiplicity but not cancer incidence. NAC, an antimucolytic agent, failed to inhibit DMBA induced mammary tumorigenesis. Surprisingly, treatment with DMBA plus PEITC, a potent inhibitor of cytochrome P450 2E1, actually increased the multiplicity of tumors relative to that observed with DMBA alone. PMID- 9212230 TI - Decreased Egr-1 expression in human, mouse and rat mammary cells and tissues correlates with tumor formation. AB - We have examined several types of tumor cell lines and shown that they invariably expressed little or no Egr-1, in contrast to their normal counterparts. We have previously shown that the expression of exogenous Egr-1 in human breast and other tumor cells markedly reduces transformed growth and tumorigenicity. We therefore hypothesized that the loss of Egr-1 expression plays a role in transformation. All human and mouse breast cancer cell lines and tumors examined had reduced Egr 1 expression compared with their normal counterparts. Reduced Egr-1 expression was also observed in 7,12-dimethylbenz(a)anthracene (DMBA)-induced rat mammary tumors, and this level increased to normal levels in tumors that regressed after tamoxifen treatment. We concluded, therefore, that loss of Egr-1 expression may play a role in the deregulation of normal growth in the tumorigenic process and that Egr-1 acts as a tumor suppressor gene. PMID- 9212231 TI - Photobleaching during and re-appearance after photodynamic therapy of topical ALA induced fluorescence in UVB-treated mouse skin. AB - Photodynamic therapy (PDT) using protoporphyrin IX (PpIX) induced by topically applied 5-aminolevulinic acid (ALA) seems a promising alternative for the treatment of superficial non-melanoma skin cancer and actinic keratosis. In this study, the kinetics of new PpIX fluorescence arising after a PDT treatment that had photobleached the original fluorescence were determined. Our purpose was to examine the feasibility of multiple irradiations, following a single topical ALA application, to increase PDT efficacy. In addition, photobleaching during PDT and the fluorescence spectra during and after PDT were studied. As a model we used hairless mice with and without UVB-induced skin lesions. ALA was applied to the skin for 4 hr. An illumination was delivered either immediately after application or 6 hr after the end of the application (at interval of maximum fluorescence). During PDT, the fluorescence of normal skin decreased at a faster rate than the fluorescence of the skin lesions. In the fluorescence study after PDT, the areas treated immediately post-application showed a fluorescence increase over time similar to that in non-treated areas on the same mice. A remarkable result was that the fluorescence of areas treated at maximum fluorescence increased, whereas the fluorescence of non-treated areas did not increase over time. With both treatment intervals the new fluorescence showed a characteristic PpIX spectrum. Our results demonstrate that a second illumination, when new PpIX fluorescence has been formed, may increase PDT efficacy after topical ALA application. This finding has been demonstrated previously for systemic ALA administration. PMID- 9212232 TI - Immunoreactivities of polyclonal and monoclonal anti-T and anti-Tn antibodies with human carcinoma cells, grown in vitro and in a xenograft model. AB - Human polyclonal, monospecific anti-T and -Tn antibodies were found to be reactive in ELISA tests with human ovarian (IGROV-1, OVCAR-3 and SKOV-3), breast (SKBr-3 and T47D)- and oral (KB)-carcinoma cell lines, but less so or non reactive with normal epithelia and fibroblasts. The direct binding radioimmunoassay, using 125I-labeled human antibodies, to the IGROV-1 cancer cells was inhibited by homologous unlabeled antibodies of the same concentration, but not by the respective immunodominant haptenic monosaccharides (Gal for T and GalNAc for Tn). Rodent ascitic monoclonal anti-T (Ca3114 and Ca3741) and anti-Tn (Ca3250, Ca3268 and Ca3638) antibodies were also reactive with the ovarian- and breast-cancer cells, as measured by FACS and ELISA tests, but to a lower extent than the polyclonal human antibodies. Both the monoclonal anti-T (Ca3741) and anti-Tn (Ca3250 and Ca3638) antibody-binding reactivities were significantly inhibited by the haptenic free monosaccharides. Addition of the above MAbs to IGROV-1 ovarian-cancer or T47D breast-cancer cells cultured in vitro resulted in significant cytological change and inhibition of the viability of the tumor cells, but not of normal epithelial breast cells. This effect on viability was shown to be complement-independent, yet it was profoundly influenced by the concentration of the serum added to the assay medium. In vivo biodistribution of the anti-T (Ca3114) and anti-Tn (Ca3638) MAbs administered i.p. to athymic IGROV 1 tumor-bearing CD1 female nude mice revealed higher 125I-labeled antibody accumulation in the tumor xenografts and in their lung tissues, as compared with other organs of the same mice tested. The above results thus suggest the feasibility of utilizing these antibodies in immunotherapy and drug targeting. PMID- 9212233 TI - Soluble and membrane isoforms of Fas/CD95 in fresh adult T-cell leukemia (ATL) cells and ATL-cell lines. AB - Fas, also designated as Apo-1 and CD95, is a cell membrane receptor (mFas) involved in apoptotic cell death. A soluble form (sFas) lacking the transmembrane domain due to alternative splicing has been isolated. Abnormal expression of sFas and mFas is likely to be involved in lymphoproliferative disorders and auto immune diseases. Adult T-cell leukemia (ATL) caused by human T-cell-leukemia virus type-1 (HTLV-1) is well known to be a T-cell neoplasm with strong mFas expression, suggesting a role of Fas in the pathology of the disease. We examined protein and mRNA expression of the 2 isoforms of Fas in fresh ATL cells and ATL cell lines. In general, mFas was strongly expressed in ATL cells, and sFas levels in sera were high, especially in malignant ATL. However, expression of the isoforms in some cases of ATL varied; there was no mFas expression on the cell surface and sFas levels were high in serum. In contrast, all ATL cell lines examined showed strong mFas expression and scarce production of sFas in the supernatant, corresponding to strong expression of full-length Fas mRNA and weak to negative expression of alternatively spliced mRNA lacking the transmembrane domain. Our findings indicate that the mode of expression of Fas isoforms in ATL cells is not always homogenous and that Fas may play a role in the malignant behavior and oncogenesis of ATL. PMID- 9212234 TI - Functional role of alpha4beta1 and alpha5beta1 integrin fibronectin receptors expressed on adriamycin-resistant MCF-7 human mammary carcinoma cells. AB - Cytofluorimetric and reverse-transcription polymerase chain reaction (RT-PCR) analysis showed that adriamycin-resistant (ADRR), but not sensitive (WT), MCF-7 human mammary carcinoma cell lines express alpha4beta1 and alpha5beta1 integrins. ADR(R) cells adhere to fibronectin (FN), and only alpha5beta1 is involved in cell adhesion to this glycoprotein, while alpha4beta1 mediates cell binding to the cellular counter-receptor VCAM-1. Proliferation assays showed that FN, but not VCAM-1, delivers a mitogenic signal to quiescent ADR(R) MCF-7 cells. The activating signal is mediated by alpha5beta1, since cell proliferation is inhibited in the presence of RGD peptide or specific antibody. Cell cycle analysis demonstrated that cell/FN interaction induces the re-entry of ADR(R) MCF 7 into S phase, and prevents them from undergoing serum deprivation-induced apoptosis. Our data suggest that the presence of alpha5beta1 on the resistant cells enables them to draw advantage from FN for both cell growth and survival. PMID- 9212235 TI - Modulations of the effector function and cytokine production of human lymphocytes by secreted factors derived from colorectal-carcinoma cells. AB - We investigated the in vitro effects of factors secreted by 3 freshly explanted human colorectal-carcinoma (CRC) cell lines on lymphocyte proliferation, IL-2 receptor expression, LAK-cell generation and cytokine secretion. We found that the supernatants of all 3 CRC cell lines inhibited T-cell proliferation in a dose dependent manner, due to the secretion of immunosuppressive factors (ISFs). In addition, the supernatants of 2 cell lines were able to inhibit LAK-cell generation and to depress IL-2R, but not HLA-DR expression, on PHA-activated T cells. Furthermore, the secretion of cytokines, i.e., IFN-gamma, IL-1beta, IL-2 and TNF-alpha, by peripheral-blood mononuclear cells (PBMC) was differently modulated by the tumor-cell supernatants, e.g., the production of IFN-gamma was reduced in normal PBMC stimulated with PHA. However, the effects induced by the supernatants were not identical: for example, factors from one CRC cell line (w25) influenced early and late events of T-cell activation and division, while 2 others (w19 and te6) contributed only to the inhibition of early events. Some biochemical properties of the ISFs were characterized. Our results suggest that colon-tumor cells can secrete ISFs, which may lead to the in vivo immunosuppression often observed in patients with these tumors. PMID- 9212236 TI - Interleukin-6 functions as an autocrine growth factor in human bladder carcinoma cell lines in vitro. AB - Interleukin (IL)-6 is reported to function as a growth factor for renal and prostatic carcinomas. We conducted the present study to define the role of IL-6 in the growth of normal and neoplastic urothelial cells. Human bladder carcinoma cell lines (253J, RT4 and T24) and primary cultured human urothelial cells derived from normal ureters were used. Recombinant human IL-6 stimulated the growth of bladder carcinoma cell lines far better than that of normal urothelial cells (p < 0.001). All carcinoma cell lines tested produced and released IL-6, whereas normal urothelial cells did so only at marginal levels. Furthermore, treatment with lipopolysaccharide derived from Escherichia coli, tumor necrosis factor-alpha or IL-1 increased IL-6 secretion by bladder carcinoma cell lines but not by normal urothelial cells. Growth of bladder carcinoma cells was significantly inhibited by anti-IL-6 neutralizing antibody or the anti-sense oligonucleotide for IL-6 cDNA. We conclude that IL-6 functions as an autocrine growth factor for bladder carcinoma cells but not for normal urothelial cells and that it may be a factor accounting for the marked enhancement of inflammation associated bladder carcinogenesis and tumor growth. PMID- 9212237 TI - Mechanism of resistance to cisplatin in a human ovarian-carcinoma cell line selected for resistance to doxorubicin: possible role of p53. AB - A possible novel mechanism of cross-resistance to cisplatin (CDDP) in the doxorubicin-resistant ovarian-cancer cell line A2780-DX3, which displays atypical multidrug resistance, is presented. A2780-DX3 is found to be more resistant than the parental line A2780 in terms of CDDP-induced cytotoxicity and apoptosis. Resistance is not related to the amount of cross-links. Topoisomerase-II (topII) protein levels were similar in both cell lines, with lower cleavage activity in A2780-DX3 cells. The parental and the doxorubicin-resistant cells expressed the same level of c-erb2, which could be implicated in CDDP resistance. bcl2 was almost undetectable in both cell lines. At the same time, we found strong induction of p53, waf-1 and bax protein levels after CDDP treatment in the A2780, but not in the A2780-DX3, cell line. Treatment of both cell lines with mitomycin C (MMC), which acts with a mechanism different from CDDP, caused equal accumulation of p53 and induction of bax. We found that A2780-DX3 cells exhibit altered cellular localization of p53 protein in comparison with A2780. A significant proportion of p53 in A2780-DX3 cells was found in the cytoplasmic compartment, and CDDP treatment induced a functional p53 protein in the nucleus of A2780 much more strongly than in A2780-DX3, which coincides with an increase of transcriptional activity of p53 in treated A2780 cells. We propose that the cross-resistance to CDDP in the A2780-DX3 cell line may be due to inactivation of a CDDP-dependent p53-accumulation pathway. PMID- 9212238 TI - Detection of Epstein-Barr virus small RNAs EBER1 and EBER2 in lymphomas of SIV infected rhesus monkeys by in situ hybridization. AB - SIV infection of macaques is a well-established animal model for studying the pathogenesis of HIV infection in humans. During the course of SIV infection, up to 40% of cynomolgus macaques (Macaca fascicularis) develop SIV-associated non Hodgkin's lymphomas. In the present study, we characterized malignant lymphomas of SIV(mac) 251/32H-infected rhesus macaques (Macaca mulatto) of our cohort in terms of clinical outcome, histopathology and EBV association. Histopathologic changes of lymphoid malignancies were classified according to the Kiel classification. For detection of the EBV-encoded small RNAs EBER1 and EBER2, a method of non-isotopic in situ hybridization was established. The presence of EBNA-2 antigens was assessed by immunohistochemistry. Seven of 43 rhesus macaques developed highly malignant B-cell lymphomas of the centroblastic, immunoblastic and Burkitt subtypes within 18-29 months post-experimental SIV infection. In situ hybridization revealed the presence of small EBER1 and -2 RNAs in 6 of 7 disease cases. EBNA-2 antigens could be demonstrated in only 4 of 7 tissue specimens. As expected, the Burkitt-type of lymphoma was negative for EBNA-2 antigen staining. In accordance with findings on SIV-associated lymphomas of cynomolgus macaques, infection with an EBV-related herpesvirus could be demonstrated in almost 90% of lymphomas in SIV-infected rhesus macaques. In contrast, the presence of EBV in lymphomas had been documented previously in only 30-40% of HIV-infected patients. Further studies should thus define the precise role of herpesvirus infection for lymphomagenesis in SIV- and HIV-induced immunodeficiency. PMID- 9212239 TI - Application of cytokine intervention for improved radio-antibody dose delivery. AB - The goal of our studies was to determine whether administration of IL-1/GM-CSF to mice could reduce radio-antibody-induced myelosuppression and allow either dose escalation of radio-antibody using 131I, 90Y or 188Re conjugated to either intact antibody or bivalent fragments or more frequent dosing with 131I-IgG. Survival, peripheral blood counts, hematopoietic tissue weight and number of marrow CFCs were used to determine the ability to dose-intensify with a single dose or to reduce the spacing between doses. In this report, we show that in the absence of cytokines, 2 cycles of 131I-IgG spaced at 28, 35, 42 and 49 days resulted in 100%, 100%, 40% and 0% lethality, respectively. In contrast, cytokine intervention reduced lethality to 45%, 20%, 0% and 0% at the same time intervals between doses. Thus, the use of cytokines permits at least a 1 week earlier redosing of 131I-IgG. Cytokine intervention also has reduced the magnitude of myelosuppression, as measured by neutropenia and thrombocytopenia, thus permitting intensification of single doses of radio-iodinated intact antibodies, bivalent fragments and 90Y-IgG by at least 30%, 50% and 25%, respectively. However, cytokines were not effective at permitting dose escalation of either 90Y F(ab')2 or 188Re-IgG. Further optimization of the dose schedule of cytokine administration needs to be explored for these 2 nuclide-antibody forms. PMID- 9212240 TI - Human gastric carcinoma transduced with the IL-2 gene: increased sensitivity to immune effector cells in vitro and in vivo. AB - We transduced a human gastric carcinoma cell line, HR, with the interleukin 2 (IL 2) gene. Stable HR transfectants secreted nanogram quantities of biologically active IL-2 and had significantly increased expression of IL-2 mRNA relative to that in parental cells. Expression of intracellular IL-2 protein was not quantitatively different in the parental and IL-2 gene-transduced cells, although the former did not secrete IL-2. Surface expression of IL-2 receptors was comparable in the parental and transduced cells at the mRNA or protein levels. Nevertheless, in vitro proliferation of IL-2 gene-transduced HR cells was significantly more rapid than that of parental cells. Both parental and IL-2 gene transduced HR cells were equally sensitive to lysis by IL-2-activated natural killer (A-NK) cells, as measured in 4 hr 51Cr-release assasys or to apoptosis induced by NK or A-NK cells, assessed in 1 hr 3H-TdR-release assays. In 24 hr MTT assays, however, the IL-2 gene-transduced cells were significantly more sensitive to these effector cells than were parental cells. Upon intrasplenic injection of 5 x 10(6) parental or transduced HR cells into nude mice, liver metastases developed. Metastases of parental HR cells killed the animals in 24 days. In contrast, metastases of the IL-2 gene-transduced HR cells became necrotic by day 14 and were found to be surrounded by murine NK cells and macrophages. Survival of nude mice injected with IL-2 gene-transduced HR cells was significantly prolonged (>50 days) relative to that of mice injected with parental HR. Thus, IL 2 gene-transduced HR cells produced sufficient amounts of functional IL-2 in vivo to be able to recruit to the tumor site and support functions of endogenous cytotoxic immune effector cells, which were responsible for regression of hepatic metastases and significant improvement of survival in these mice. PMID- 9212241 TI - Reduced folate carrier gene (RFC1) expression and anti-folate resistance in transfected and non-selected cell lines. AB - Methotrexate transport deficiency due to decreased reduced folate carrier (RFC) activity has been observed in several cell lines selected for resistance to methotrexate (MTX). Since MTX resistance is multifactorial, however, it is difficult to quantify the relative importance of changes in RFC activity in selected cell lines and even more so to determine the relative contribution of naturally occurring RFC activity in the MTX sensitivity of non-selected cell lines. We examined the role of RFC in MTX resistance by studying a transport deficient cell line transfected with the gene for human RFC, RFC1, and by correlating relative RFC1 expression with MTX and trimetrexate (TMTX) growth inhibition (GI50) in a panel of cell lines used in the NCI Anticancer Drug Screen. Clones of transport-deficient, MTX-resistant ZR-75-1 human breast cancer cells (MTX(R) ZR-75-1) transfected with RFC1 were 250-fold more sensitive to MTX and 300-fold more resistant to TMTX than control cell clones, showing that restoration of RFC activity has a significant impact on MTX and TMTX cytotoxicity. We also surveyed 40 of the 60 cell lines in the NCI drug screen panel for RFCI RNA levels by a quantitative RT-PCR assay. RFCI RNA levels varied over a range of 15-fold, with only 1 cell line found to be null in expression. Using data from the 6-day drug exposure assay, RFC1 correlated positively with MTX and negatively with TMTX cytotoxicity. As predicted by transfection studies, the calculated difference between MTX and TMTX potency was even more strongly correlated with RFC1 RNA levels of the cell lines. In addition, compounds in the NCI Anticancer Drug Screen database with cytotoxicity profiles which correlated with RFC1 RNA levels or with the calculated difference in MTX-TMTX potency were examined for MTX uptake inhibition and cytotoxicity in the RFC1-transfected MTX(R) ZR-75-1 cell line. Overall, our data demonstrate the importance of RFC1 in MTX resistance both as a transgene and as a constitutively expressed gene in non selected cell lines. PMID- 9212242 TI - Ectopic expression of target genes may represent an inherent limitation of RT-PCR assays used for micrometastasis detection: studies on the epithelial glycoprotein gene EGP-2. AB - Our objective was to develop and study the feasibility of a quantitative, nested reverse-transcription polymerase chain reaction (RT-PCR) assay for detection of micrometastatic, epithelial tumor cells using the epithelial glycoprotein EGP-2 gene as a target. Several carcinoma cell lines and peripheral blood samples of 10 healthy volunteers were screened for levels of EGP-2 mRNA. The assay included EGP 2 competitor molecules, carrying an internal deletion, that had been titrated by limiting dilution. Seven carcinoma cell lines showed a wide spectrum of EGP-2 mRNA expression levels, with the highest values (20-100 molecules/cell) seen in 3 breast-cancer cell lines. Unexpectedly, a consistent low level of EGP-2 mRNA expression (0.0004 molecules/cell) was observed in peripheral blood mononuclear cells, probably representing ectopic non-functional expression. Because of this background level, spiking experiments with T47D breast-carcinoma cells added to blood mononuclear cells exhibited a detection limit that was not better than approximately one tumor cell in 2 x 10(4) normal cells. Together with the considerable variation of EGP-2 transcript levels that is observed in different carcinoma cell lines, the extent of expression in normal blood cells would prevent a reliable estimation of low numbers of carcinoma cells in clinical samples. A similar situation might well apply for other target genes. This emphasizes the need for a critical evaluation of the different steps involved in the methods used for RT-PCR detection of micrometastatic tumor cells. PMID- 9212243 TI - Expression of transgenic carcinoembryonic antigen (CEA) in tumor-prone mice: an animal model for CEA-directed tumor immunotherapy. AB - Carcinoembryonic antigen (CEA) is a tumor marker for the most common forms of adenocarcinomas. We have previously described C57BL/6 mice transgenic for the complete human CEA gene. Compared with humans, these mice reveal a conserved spatiotemporal CEA expression pattern. To establish animal models for CEA targeted tumor immunotherapy, we have crossed CEA transgenic mice with mice that are genetically predisposed to tumor development. These immunocompetent animals should allow optimization of immunotherapy strategies for maximal destruction of tumor tissues with minimal damage to CEA-expressing normal tissues. To develop a breast tumor model, CEA transgenic mice were cross-bred with mice transgenic for the rat neu protooncogene controlled by the mouse mammary tumor virus long terminal repeat. Female offspring developed poorly differentiated breast tumors, none of which, however, expressed CEA. As a model for colorectal tumors, mice bearing a mutation in the Apc gene (Min mice) and the CEA transgene developed multiple intestinal adenomas with strong CEA expression in all tumor cells. CEA expression had no significant effect on tumor growth. Occasional, well differentiated breast adenocarcinomas in female offspring expressed CEA focally in tumor cells lining pseudolumina. Cross-breeding Apc(Min/+) mice with neu transgenic mice did not reveal a synergistic effect on the kinetics of breast tumor formation. Finally, CEA transgenic mice crossbred with mice transgenic for the SV40 large T antigen regulated by the surfactant protein-C promoter, developed multiple lung adenocarcinomas that revealed a mosaic CEA expression pattern. PMID- 9212244 TI - Processing of interaural intensity differences in the LSO: role of interaural threshold differences. AB - Cells in the lateral superior olive (LSO) are known to be sensitive to interaural intensity differences (IIDs) in that they are excited by IIDs that favor the ipsilateral ear and inhibited by IIDs that favor the contralateral ear. For each LSO neuron there is a particular IID that causes a complete inhibition of discharges, and the IID of complete inhibition varies from neuron to neuron. This variability in IID sensitivity among LSO neurons is a key factor that allows for the coding of a variety of IIDs among the population of cells. A fundamental question concerning the coding of IIDs is: how does each cell in the LSO derive its particular IID sensitivity? Although there have been a large number of neurophysiological studies on the LSO, this question has received little attention. Indeed, the only reports that have directly addressed this question are those of Reed and Blum, who modeled the binaural properties of LSO neurons and proposed that the IID at which discharges are completely suppressed should correspond to the difference in threshold between the excitatory, ipsilateral and inhibitory, contralateral inputs that innervate each LSO cell. The main purpose of this study was to test the threshold difference hypothesis proposed by Reed and Blum by recording responses to monaural stimulation and to IIDs from single cells in the LSO of the mustache bat. Our results show that although the IID sensitivities of some LSO cells correspond to the difference in threshold between the excitatory and inhibitory ears, in the majority of cells the difference in thresholds did not correspond to the cell's IID sensitivity. The results lead us to propose two models to account for IID sensitivities. One model is similar to that proposed by Reed and Blum and emphasizes differences in the thresholds of the excitatory and inhibitory inputs. This model accounts for the minority of cells in which the IID of complete inhibition corresponded to the difference in threshold of the inputs from the two ears. The other model, which accounts for the cells in which the IID of complete inhibition did not correspond to the difference in the thresholds of the inputs from the two ears (the majority of cells), places emphasis on differences in latencies of the excitatory and inhibitory inputs. The models incorporate features that are concordant with the known properties of the neurons that project to the LSO and together can account for the diversity of IID sensitivities among the population of LSO neurons. PMID- 9212245 TI - Encoding of binocular disparity by complex cells in the cat's visual cortex. AB - To examine the roles that complex cells play in stereopsis, we have recorded extracellularly from isolated single neurons in the striate cortex of anesthetized paralyzed cats. We measured binocular responses of complex cells using a comprehensive stimulus set that encompasses all possible combinations of positions over the receptive fields for the two eyes. For a given position combination, stimulus contrast could be the same for the two eyes (2 bright or 2 dark bars) or opposite (1 bright and 1 dark). These measurements provide a binocular receptive field (RF) profile that completely characterizes complex cell responses in a joint domain of left and right stimulus positions. Complex cells typically exhibit a strong selectivity for binocular disparity, but are only broadly selective for stimulus position. For most cells, selectivity for disparity is more than twice as narrow as that for position. These characteristics are highly desirable if we assume that a disparity sensor should exhibit position invariance while encoding small changes in stimulus depth. Complex cells have nearly identical binocular RFs for bright and dark stimuli as long as the sign of stimulus contrast is the same for the two eyes. When stimulus contrast is opposite, the binocular RF also is inverted such that excitatory subregions become suppressive. We have developed a disparity energy model that accounts for the behavior of disparity-sensitive complex cells. This is a hierarchical model that incorporates specific constraints on the selection of simple cells from which a complex cell receives input. Experimental data are used to examine quantitatively predictions of the model. Responses of complex cells generally agree well with predictions of the disparity energy model. However, various types of deviations from the predictions also are found, including a highly elongated excitatory region beyond that supported by a single energy mechanism. Complex cells in the visual cortex appear to provide a next level of abstraction in encoding information for stereopsis based on the activity of a group of simple-type subunits. In addition to exhibiting narrow disparity tuning and position invariance, these cells seem to provide a partial solution to the stereo correspondence problem that arises in complex natural scenes. Based on their binocular response properties, these cells provide a substantial reduction in the complexity of the correspondence problem. PMID- 9212246 TI - Multiple effects of serotonin on membrane properties of trigeminal motoneurons in vitro. AB - Intracellular recordings from guinea pig trigeminal motoneurons (TMNs) in brain stem slices were used to determine the underlying ionic mechanisms responsible for our previously demonstrated enhancement of TMN excitability during jaw movements by serotonin (5-HT). 5-HT (0.5-100 microM) depolarized motoneurons and increased input resistance in the majority of neurons tested. Additionally, 5-HT reduced the amplitude of the postspike medium-duration afterhyperpolarization, decreased the current threshold for maintained spike discharge, and increased the maximum slope of the steady-state spike frequency-current relationship. Under voltage clamp, from holding potentials close to resting potential, 5-HT produced an inward current and a decrease in instantaneous slope conductance, suggesting a reduction in a resting K+ leak conductance (I(leak)). The instantaneous current voltage (I-V) relationship for the inward 5-HT current (I(5-HT)) was linear throughout most of the voltage range tested. However, the steady-state I-V relationship showed some degree of inward rectification at potentials starting around -70 mV. The mean reversal potential for the instantaneous I(5-HT) was 86.2 +/- 4.5 (SE) mV (n = 9), a value slightly negative to the predicted potassium equilibrium potential of -82 mV in these neurons. In the presence of 2 mM Ba2+, 5-HT application did not produce a further reduction in input conductance, but did expose a Ba2+-insensitive residual inward current that was resistant to Cs+ application. The instantaneous I-V relationship during 5-HT application in the presence of Ba2+ was shifted downward and parallel to control, suggesting that Ba2+ and 5-HT block the same resting I(leak). The residual Ba2+- and Cs+-insensitive component of the total inward I(5-HT) was voltage independent and was blocked when the extracellular Na+ was replaced by choline, suggesting that the predominant charge carrier for this residual current is Na+. 5-HT enhanced a hyperpolarization-activated cationic current, I(h). In the presence of Ba2+, the time course of I(5-HT) resembled that of I(h) and showed a similar voltage dependence that was blocked by extracellular Cs+ (1-3 mM). The effects of 5-HT on membrane potential, input resistance, and I(h) were partially mimicked by 5-HT2 agonists and suppressed by 5-HT2 antagonists. It is concluded that 5-HT enhances TMN membrane excitability through modulation of multiple intrinsic membrane conductances. This provides for a mechanism(s) to fine tune the input output discharge properties of these neurons, thus providing them with greater flexibility in output in response to time-varying synaptic inputs during various movements of the jaw. PMID- 9212247 TI - Primary- and secondary-like jaw-muscle spindle afferents have characteristic topographic distributions. AB - Single jaw-muscle spindle afferent axons were characterized physiologically and intracellularly stained to determine whether particular physiological types of spindle afferent show distinctive morphologies. Microelectrodes filled with either horseradish peroxidase (HRP) or biotinamide (Neurobiotin) were advanced into the mesencephalic trigeminal nucleus (Vme) in anesthetized rats. Intracellular recordings then were characterized by their response: to palpation of the jaw muscles; when pressure was applied to the teeth and during passive ramp and hold and sinusoidal jaw movement. Seventy-one afferents were characterized physiologically and injected with HRP; an additional 61 afferents were typed and injected with biotinamide. The response of 43 stained neurons was recorded in the presence of suxamethonium. The major projection areas of these afferents were the: trigeminal motor nucleus (Vmo); region dorsal to Vmo; reticular formation, spinal trigeminal nucleus, superior cerebellar peduncle and Vme. One afferent type was modulated strongly during stretching of the jaw elevator muscles. Based on their high sensitivity during stretching of the jaw muscles and/or their silencing during the release phase of muscle stretch, these afferents were classified as primary-like spindle afferents. These afferents projected most strongly to Vmo. A second type of afferent was modulated only modestly during stretching of the jaw-elevator muscles. These tonic afferents were classified as secondary-like spindle afferents because of their low dynamic sensitivity during ramp muscle stretch and their continued discharge during the release phase of muscle stretch. Secondary-like afferents projected most strongly to the region dorsal to Vmo. Boutons (n = 3,834) from 11 afferents were studied in detail. Secondary-like afferents had statistically larger boutons within Vmo. In both secondary- and primary-like spindle afferents, only a small number of boutons were associated closely with the somata and proximal dendrites of trigeminal motoneurons. In these cases, however, two to five boutons appeared to contact individual motoneurons, implying multiple monosynaptic inputs to a selective subset of jaw-elevator motoneurons. Some "giant" boutons were present dorsal to Vmo and in Vme. These results demonstrate that dynamically sensitive and nondynamically sensitive jaw-elevator muscle spindle afferents project preferentially to different regions. Primary-like spindle afferents are capable of providing feedback related to the dynamic phases of muscle stretch and project most heavily to Vmo. Secondary-like spindle afferents can transmit a feedback signal associated with muscle length and project most strongly to the supratrigeminal region. Both types of afferent have projections caudal to Vmo that may serve longer latency jaw-muscle stretch reflexes and/or the projection of proprioceptive information to the thalamus and cerebellum. PMID- 9212248 TI - Regularity of firing of neurons in the inferior colliculus. AB - The spike discharge regularity of 254 tonically firing units in the inferior colliculus (IC) of the anesthetized guinea pig was studied in response to tones presented at best frequency (BF) to the ear contralateral to the recorded IC. Regularity of firing was measured by calculating the coefficient of variation (CV) as a function of time over the course of a unit's response. Two hundred and fifteen units (56 under urethan and 159 under chloralose anesthesia) in the central nucleus of the IC (CNIC) were studied in detail. In response to tones at 15-25 dB above threshold, 80% of units in the urethan sample fired regularly (CV < 0.5) during their sustained response, and 46% were highly regular (CV < or = 0.35). For chloralose the values were 68% and 23%, respectively. Units recorded under urethan were significantly more regular than those recorded under chloralose. For units in the sample with a measurable onset CV, 63% were regular and 44% highly regular under urethan, and 73% were regular and 54% highly regular under chloralose. The units' peristimulus time histogram (PSTH) patterns were classified into subdivisions of four categories: choppers [9%: chop-sustained (Cs), chop-onset (Co)]; pausers [42%: pauser-chop-sustained (P/Cs), pauser-chop onset (P/Co), pauser-no-chop]; ON-sustained (43%: primary-type, L-type, h-type); and sustained (6%). The presence of chopping was a reliable predictor of regularity: Cs and P/Cs units were highly regular throughout their response, whereas Co and P/Co units were highly regular at onset and became less regular. Some units in the other PSTH categories were highly regular despite the absence of chopping, and units with virtually identical PSTHs showed very different sustained CVs. Regularity was measured as a function of firing rate in 71 units. In 23%, regularity remained constant when firing rate changed with stimulus level. Forty-six percent fired more regularly as firing rate increased, 8% fired less regularly, and 23% of units showed no consistent relationship between CV and firing rate. Regularity did not correlate with the neurons' frequency response areas or BFs. Regular firing was also found in a smaller sample of units recorded in cortices surrounding the CNIC. We conclude that regular firing is a characteristic feature of most neurons in the IC. Regularity is a specific feature correlated with four PSTH types (Cs, Co, P/Cs, and P/Co). Other PSTH types may or may not exhibit regularity. PMID- 9212249 TI - Sympathetically correlated activity of dorsal horn neurons in spinally transected rats. AB - In mammals with an intact neuraxis, most sympathetic nerve activity is generated by brain stem systems. Therefore these systems have attracted much more attention than spinal systems that generate excitatory inputs to sympathetic preganglionic neurons. The purpose of this study was to determine whether, within hours of C1 spinal cord transection, spinal dorsal horn neurons (DHNs) play a role in generating sympathetic nerve activity. Experiments were conducted in chloralose anesthetized rats. We recorded renal sympathetic nerve activity (RSNA) in the left renal nerve, and we recorded the activity of neurons located in the left dorsal horn at T2, T8, T10, T13, and L2. We also recorded the activity of neurons in the right dorsal horn at T10. The somatic fields and cutaneous modalities of most neurons were determined. Spike-triggered averaging was used to determine relationships between the ongoing activity of DHNs and ongoing RSNA. In the left dorsal horn, bursts of ongoing activity of 16% of DHNs at T8 and 43% of DHNs at T10 were positively correlated with bursts of ongoing RSNA at latencies of 59 +/- 8 (SE) ms. At no other level on the left side, nor in the T10 segment on the right side, was the activity of DHNs correlated with RSNA. DHNs with activity correlated with RSNA were located only in dorsal horn laminae III-V. Deeper laminae were not investigated in these experiments. The activity of all sympathetically correlated DHNs exhibited bursts of action potentials with interspike intervals of < 10 ms. All but one of the sympathetically correlated DHNs exhibited wide-dynamic-range modalities. The modalities of sympathetically uncorrelated neurons were more heterogeneous. Brief (5-10 s) noxious cutaneous stimulation of mid- and lower thoracic dermatomes on the left side excited all sympathetically correlated DHNs and simultaneously increased RSNA. The excitatory cutaneous fields of sympathetically correlated neurons were circumscribed by the excitatory fields for RSNA. The excitatory cutaneous fields of some sympathetically uncorrelated DHNs extended beyond the excitatory fields for RSNA. Noxious cutaneous stimulation of the extremities on the left side that decreased RSNA simultaneously decreased the activity of all sympathetically correlated DHNs. These data provide electrophysiological evidence that, in spinally transected rats, a population of DHNs may generate or convey excitatory input to renal sympathetic preganglionic neurons. PMID- 9212250 TI - Excitation of the brain stem pedunculopontine tegmentum cholinergic cells induces wakefulness and REM sleep. AB - Considerable evidence suggests that brain stem pedunculopontine tegmentum (PPT) cholinergic cells are critically involved in the normal regulation of wakefulness and rapid eye movement (REM) sleep. However, much of this evidence comes from indirect studies. Thus, although involvement of PPT cholinergic neurons has been suggested by numerous investigations, the excitation of PPT cholinergic neurons causal to the behavioral state of wakefulness and REM sleep has never been directly demonstrated. In the present study we examined the effects of three different levels of activation of PPT cholinergic cells in wakefulness and sleep behavior. The effects of glutamate on the activity of PPT cholinergic cells were studied by microinjection of one of the three different doses of L-glutamate (0.3, 1.0, and 3.0 microg) or saline (vehicle control) into the PPT cholinergic cell compartment while quantifying the effects on wakefulness and sleep in free moving chronically instrumented cats. All microinjections were made during wakefulness and were followed by 4 h of recording. Polygraphic records were scored for wakefulness, slow-wave sleep states 1 and 2, slow-wave sleep with pontogeniculooccipital waves, and REM sleep. Dependent variables quantified after each microinjection included the percentage of recording time spent in each state, the latency to onset of REM sleep, the number of episodes per hour for REM sleep, and the duration of each REM sleep episode. A total of 48 microinjections was made into 12 PPT sites in six cats. Microinjection of 0.3- and 1.0-microg doses of L-glutamate into the cholinergic cell compartment of the PPT increased the total amount of REM sleep in a dose-dependent manner. Both doses of L glutamate increased REM sleep at the expense of slow-wave sleep but not wakefulness. Microinjection of 3.0 microg L-glutamate kept animals awake for 2-3 h by eliminating slow-wave and REM sleep. The results show that the microinjection of the excitatory amino acid L-glutamate into the PPT cholinergic cell compartments can increase wakefulness and/or REM sleep depending on the L glutamate dosage. These findings unambiguously confirm the hypothesis that the excitation of the PPT cholinergic cells is causal to the generation of wakefulness and REM sleep. PMID- 9212251 TI - Warm-sensitive afferent splanchnic C-fiber units in vitro. AB - Receptive fields of 41 slowly conducting sensory fibers were located using a thermal (warm) search stimulus in an in vitro splanchnic nerve-mesentery preparation. Warm-sensitive receptive fields were punctate and were densest in the region surrounding the prevertebral ganglia, an area with prominent deposits of brown adipose tissue, where the abdominal aorta branches into the major trunks supplying the abdominal viscera. Impulse activity was recorded while applying a warm stimulus to identified receptive fields (RFs). The warm stimulus consisted of a warming ramp (10-15 degrees C in 1-2 s to a 42-49 degrees C peak temperature) followed by a 10- to 30-s period during which the RF was maintained at this peak temperature (plateau phase). Eighty percent (33/41) of warm sensitive units responded to warming with discharge comprising both a phasic and a tonic component (slowly adapting warm-sensitive, or SA-W, units). The remainder (8/41) responded with only phasic discharge (rapidly adapting warm-sensitive, or RA-W, units). Units' adaptation characteristics were consistent from trial to trial and when applying stimuli from different positions. Fifty percent of SA-W units (8/16) and 17% of RA-W units (1/6) were activated by transient exposure to 9-90 nM bradykinin (BK). Twenty-seven percent (9/33) of SA-W units and 12% (1/8) of RA-W units were activated by probing their RF with von Frey hairs with bending forces < 10 mN (approximately 1 g equivalent mass). An additional five SA-W units tested were activated by strong mechanical stimuli (compression with a metal probe or firm stretching). No BK-responsive warm-sensitive units were activated by von Frey probing < 10 mN, but two (both SA-W) responded to strong mechanical stimuli. In six SA-W units and one RA-W unit, the number of impulses evoked by warming approximately 5 min after exposure to BK was > 2 SD greater than the mean pre-BK response, indicating sensitization. This sensitization was transient, the response to warming returning to within one standard deviation of the pretrial mean or less over the course of the next 5-10 min. Changes in background activity, mechanical sensitivity, BK sensitivity, and BK-induced sensitization were noted in various splanchnic units over the course of prolonged observations, suggesting that these indices may not reliably distinguish unit type, but instead may indicate the functional state of the sense organ. Splanchnic neurons responsive to the intense warming used in the present in vitro experiments may participate in the cardiovascular responses observed in vivo in heat-stressed rats. The dense distribution of warm-receptive fields in the vicinity of the celiac-superior mesenteric ganglionic complex is consistent with the localization of splanchnic thermosensitive units previously noted in vivo in the rabbit. PMID- 9212252 TI - Sacculocollic reflex arcs in cats. AB - Neuronal connections and pathways underlying sacculocollic reflexes were studied by intracellular recordings from neck extensor and flexor motoneurons in decerebrate cat. Bipolar electrodes were placed within the left saccular nerve, whereas other branches of the vestibular nerve were removed in the inner ear. To prevent spread of stimulus current to other branches of the vestibular nerve, the saccular nerve and the electrodes were covered with warm semisolid paraffin Vaseline mixture. Saccular nerve stimulation evoked disynaptic (1.8-3.0 ms) excitatory postsynaptic potentials (EPSPs) in ipsilateral neck extensor motoneurons and di- or trisynaptic (1.8-4.0 ms) EPSPs in contralateral neck extensor motoneurons, and di- and trisynaptic (1.7-3.6 ms) inhibitory postsynaptic potentials (IPSPs) in ipsilateral neck flexor motoneurons and trisynaptic (2.7-4.0 ms) IPSPs in contralateral neck flexor motoneurons. Ipsilateral inputs were about twice as strong as contralateral ones to both extensor and flexor motoneurons. To determine the pathways mediating this connectivity, the lateral part of the spinal cord containing the ipsilateral lateral vestibulospinal tract (i-LVST) or the central part of the spinal cord containing the medial vestibulospinal tracts (MVSTs) and possibly reticulospinal fibers (RSTs) were transected at the caudal end of the C1 segment. Subsequent renewed intracellular recordings following sacculus nerve stimulation indicated that the pathway from the saccular nerve to the ipsilateral neck extensor motoneurons projects though the i-LVST, whereas the pathways to the contralateral neck extensors and to the bilateral neck flexor motoneurons descend in the MVSTs/RSTs. Our data show that sacculo-neck reflex connections display a qualitatively bilaterally symmetrical innervation pattern with excitatory connections to both neck extensor motoneuron pools, and inhibitory connections to both neck flexor motoneuron pools. This bilateral organization contrasts with the unilateral innervation scheme of the utriculus system. These results suggest a different symmetry plane along which sacculus postural reflexes are organized, thus supplementing the reference planes of the utriculus system and allowing the gravistatic system to represent all three translational spatial degrees of freedom. We furthermore suggest that the sacculocollic reflex plays an important role in maintaining the relative position of the head and the body against the vertical linear acceleration of gravity. PMID- 9212253 TI - Noradrenergic regulation of synaptic plasticity in the hippocampal CA1 region. AB - The effects of norepinephrine (NE) and related agents on long-lasting changes in synaptic efficacy induced by several patterns of afferent stimuli were investigated in the CA1 region of rat hippocampal slices. NE (10 microM) showed little effect on the induction of long-term potentiation (LTP) triggered by theta burst-patterned stimulation, whereas it inhibited the induction of long-term depression (LTD) triggered by 900 pulses of 1-Hz stimulation. In nontreated slices, 900 pulses of stimuli induced LTD when applied at lower frequencies (1-3 Hz), and induced LTP when applied at a higher frequency (30 Hz). NE (10 microM) caused a shift of the frequency-response relationship in the direction preferring potentiation. The effect of NE was most prominent at a stimulus frequency of 10 Hz, which induced no changes in control slices but clearly induced LTP in the presence of NE. The facilitating effect of NE on the induction of LTP by 10-Hz stimulation was blocked by the beta-adrenergic receptor antagonist timolol (50 microM), but not by the alpha receptor antagonist phentolamine (50 microM), and was mimicked by the beta-agonist isoproterenol (0.3 microM), but not by the alpha1 agonist phenylephrine (10 microM). The induction of LTD by 1-Hz stimulation was prevented by isoproterenol but not by phenylephrine, indicating that the activation of beta-receptors is responsible for these effects of NE. NE (10 microM) also prevented the reversal of LTP (depotentiation) by 900 pulses of 1-Hz stimulation delivered 30 min after LTP induction. In contrast to effects on naive (nonpotentiated) synapses, the effect of NE on previously potentiated synapses was only partially mimicked by isoproterenol, but fully mimicked by coapplication of phenylephrine and isoproterenol. In addition, the effect of NE was attenuated either by phentolamine or by timolol, indicating that activation of both alpha1 and beta-receptors is required. These results show that NE plays a modulatory role in the induction of hippocampal synaptic plasticity. Although beta-receptor activation is essential, alpha1 receptor activation is also necessary in determining effects on previously potentiated synapses. PMID- 9212254 TI - Late sodium channel openings underlying epileptiform activity are preferentially diminished by the anticonvulsant phenytoin. AB - Late openings of sodium channels were observed in outside-out patch recordings from hippocampal neurons in culture. In previous studies of such neurons, a persistent sodium current appeared to underlie the ictal epileptiform activity. All the channel currents were blocked by tetrodotoxin. In addition to the transient openings of sodium channels making up the peak sodium current, there were two types of late channel openings: brief late and burst openings. These late channel openings occurred throughout voltage pulses that lasted 750 ms, producing a persistent sodium current. At -30 mV, this current was 0.4% of the peak current. The late channel openings occurred throughout the physiological range of trans-membrane voltages. The anticonvulsant phenytoin reduced the late channel openings more than the peak currents. The effect on the persistent current was greatest at more depolarized voltages, whereas the effect on peak currents was not substantially voltage dependent. In the presence of 60 microM phenytoin, peak sodium currents at -30 mV were 40-41% of control, as calculated using different methods of analysis. Late currents were 22-24% of control. Phenytoin primarily decreased the number of channel openings, with less effect on the duration of channel openings and no effect on open channel current. This set of findings is consistent with models in which phenytoin binds to the inactivated state of the channel. The preferential effect of phenytoin on the persistent sodium current suggests that an important pharmacological mechanism for a sodium channel anticonvulsant is to reduce late openings of sodium channels, rather than reducing all sodium channel openings. We hypothesize that pharmacological interventions that are most selective in reducing late openings of sodium channels, while leaving early channel openings relatively intact, will be those that produce an anticonvulsant effect while interfering minimally with normal function. PMID- 9212255 TI - Adenosine modulation of calcium currents and presynaptic inhibition of GABA release in suprachiasmatic and arcuate nucleus neurons. AB - Adenosine modulation of calcium channel currents and synaptic gamma-aminobutyrate (GABA) release was investigated with whole cell voltage-clamp recordings in rat suprachiasmatic nucleus (SCN) and arcuate nucleus cultures (n = 94). In SCN cultures, approximately 70% of the neurons showed a reversible inhibition of whole cell barium currents on the application of adenosine or its analogues. Adenosine at 1 microM reduced the amplitude of the barium currents by approximately 27%. In contrast to the significant reduction in the amplitude, the rising and decaying phases of the barium currents, and the inverted bell shape of the current-voltage curve of the barium currents, were not changed by adenosine. The adenosine A1 receptor agonist N6-cyclopentyladenosine (CPA; 100 nM) and the adenosine A2 receptor agonist N6-[2-(3,5-dimethoxyphenyl)-ethyl]adenosine (DPMA; 100 nM) inhibited the barium currents by 21% and 16%, respectively, in SCN neurons, indicating both A1 and A2 receptor actions. The A1 receptor antagonist 8 cyclopentyl-1,3-dipropylxanthine (100 nM) significantly reduced the effect of CPA but did not change the effect of DPMA on the barium currents. In the presence of tetrodotoxin to block action potentials, the frequency, but not the amplitude, of miniature inhibitory postsynaptic currents was significantly reduced (46%) by 1 microM adenosine, suggesting a presynaptic mechanism of adenosine action. In support of this suggestion, the postsynaptic GABA receptor responses were not influenced by 1 microM adenosine in the majority of SCN neurons. Most solitary self-innervating SCN neurons in microisland cultures were GABAergic. In these cells, the evoked autaptic GABA release (inhibitory postsynaptic current) was significantly inhibited by adenosine (37%), CPA (27%), and DPMA (28%), indicating that both A1 and A2 receptors were present in presynaptic axons. Similar to the effect in SCN neurons, adenosine inhibited both barium currents and GABA release in arcuate neurons. The reduction of whole cell barium currents by adenosine (1 microM), CPA (100 nM), and DPMA (100 nM) was 24, 17, and 19%, respectively. In solitary self-innervating arcuate neurons, adenosine inhibited the evoked GABA release (inhibitory postsynaptic current) by approximately 48%. We conclude that both adenosine A1 and A2 receptors are present in the SCN and arcuate nucleus of the hypothalamus. Adenosine inhibits calcium currents and presynaptically reduces inhibitory GABA neurotransmission. PMID- 9212257 TI - Neuronal encoding of sound direction in the auditory midbrain of the rainbow trout. AB - Acoustical stimulation causes displacement of the sensory hair cells relative to the otoliths of the fish inner ear. The swimbladder, transforming the acoustical pressure component into displacement, also contributes to the displacement of the hair cells. Together, this (generally) yields elliptical displacement orbits. Alternative mechanisms of fish directional hearing are proposed by the phase model, which requires a temporal neuronal code, and by the orbit model, which requires a spike density code. We investigated whether the directional selective response of auditory neurons in the midbrain torus semicircularis (TS; homologous to the inferior colliculus) is based on spike density and/or temporal encoding. Rainbow trout were mounted on top of a vibrating table that was driven in the horizontal plane to simulate sound source direction. Rectilinear and elliptical (or circular) motion was applied at 172 Hz. Generally, responses to rectilinear and elliptical/circular stimuli (irrespective of direction of revolution) were the same. The response of auditory neurons was either directionally selective (DS units, n = 85) or not (non-DS units, n = 106). The average spontaneous discharge rate of DS units was less than that of non-DS units. Most DS units (70%) had spontaneous activities < 1 spike per second. Response latencies (mode at 18 ms) were similar for both types of units. The response of DS units is transient (19%), sustained (34%), or mixed (47%). The response of 75% of the DS units synchronized to stimulus frequency, whereas just 23% of the non-DS responses did. Synchronized responses were measured at stimulus amplitudes as low as 0.5 nm (at 172 Hz), which is much lower than for auditory neurons in the medulla of the trout, suggesting strong convergence of VIIIth nerve input. The instant of firing of 42% of the units was independent of stimulus direction (shift <15 degrees), but for the other units, a direction dependent phase shift was observed. In the medial TS spatial tuning of DS units is in the rostrocaudal direction, whereas in the lateral TS all preferred directions are present. On average, medial DS units have a broader directional selectivity range, are less often synchronized, and show a smaller shift of the instant of firing as a function of stimulus direction than lateral DS units. DS response characteristics are discussed in relation to different hypotheses. We conclude that the results are more in favor of the phase model. PMID- 9212256 TI - Responses to glutamate in rat taste cells. AB - We studied taste transduction in sensory receptor cells. Specifically, we examined the actions of glutamate, a significant taste stimulus, on the membrane properties of taste cells by applying whole cell patch-clamp techniques to cells in rat taste buds isolated from foliate and vallate papillae. In 55 of 91 taste cells, bath-applied glutamate, at concentrations that elicit taste responses in the intact animal (10-20 mM), produced one of two different responses when the cell membrane was held near its presumed resting potential, -85 mV. "Sustained" glutamate responses were observed in the majority of taste cells (51 of 55) and consisted of an outward current (reduction of the maintained inward current). Sustained glutamate responses were voltage dependent, were decreased by membrane depolarization, and were accompanied by a reduction in membrane conductance. An analysis of the reversal potential of sustained responses in different ionic conditions and the effect of ion substitutions suggested that the currents were carried by cations. The data suggest that sustained responses are mediated by the closure of nonselective cation channels. Other taste cells (4 of 55) responded to glutamate with a transient inward current--so-called "transient" responses. Transient glutamate responses were voltage dependent and Na+ dependent, and appeared to be generated by nonspecific cation channels activated by glutamate. L(+)-2-amino-4-phosphonobutyric acid (L-AP4), a specific agonist of a metabotropic glutamate receptor (mGluR4) recently identified in rat taste cells and believed to be involved in taste transduction, mimicked the sustained glutamate responses. These findings indicate that glutamate, at concentrations at or slightly above threshold for taste in rats, produces two different membrane currents. The properties of these two responses suggest that there may be two different sets of nonspecific cation channels in taste cells, one closed by glutamate (sustained response) and the other opened (transient response). Our findings on the effect of L-AP4 suggest that the sustained response is the membrane mechanism mediating, at least in part, taste transduction for glutamate. PMID- 9212258 TI - Ionic currents of isolated retinal pacemaker neurons: projected daily phase differences and selective enhancement by a phase-shifting neurotransmitter. AB - The eye of Aplysia expresses a robust circadian rhythm of neuronal activity. We dissociated the retinal cells in primary culture and studied isolated pacemaker neurons to identify ionic currents that may have roles in the circadian clock mechanism. Individual neurons were studied with perforated-patch whole cell recording techniques in current- and voltage-clamp modes. Pacemaker neurons had resting potentials near -40 mV and, if neurites had grown out, produced spontaneous action potentials in darkness at <1 Hz. Depolarizing current injections increased the rate of action potential firing. Hyperpolarizing current injections were followed by slowly decaying (1-3 s) afterhyperpolarizations. Four ionic currents were characterized under voltage-clamp, including a Ca current (I(Ca)), a voltage-gated potassium current (I(KV)), an A current (I(A)), and a hyperpolarization-activated Cl current (I(Cl)). I(Cl) was only seen using Cl(-) filled electrodes when high concentrations of Cl- diffused from the electrode and is therefore unlikely to be important under physiological conditions. The magnitude of I(KV) was significantly larger during the projected zeitgeber predawn phase than during the postdawn phase, whereas the magnitude of I(A) was constant at these circadian phases, suggesting that only I(KV) is controlled by the circadian clock. Serotonin increased I(KV) by 29%, consistent with reports that serotonin suppresses optic nerve activity and phase shifts the circadian rhythm recorded from the intact eye. The enhancement of I(KV) likely contributes to membrane hyperpolarization, and it may be required for phase shifting. The phase-dependent changes in I(KV) provide evidence that each retinal pacemaker neuron contains a circadian clock, but confirmation must await further recordings made from individual pacemaker neurons that are isolated completely from all other cells in primary culture. From the present experiments, it appears that I(KV) is controlled by the circadian clock, in part, and it may be a required element in the pathway that is activated during serotonin-induced phase shifts. PMID- 9212259 TI - Responses of neurons in the inferior colliculus to binaural masking level difference stimuli measured by rate-versus-level functions. AB - The psychophysical detection threshold of a low-frequency tone masked by broadband noise is reduced by < or = 15 dB by inversion of the tone in one ear (called the binaural masking level difference: BMLD). The contribution of 120 low frequency neurons (best frequencies 168-2,090 Hz) in the inferior colliculus (ICC) of the guinea pig to binaural unmasking of 500-Hz tones masked by broadband noise was examined. We measured rate-level functions of the responses to identical signals (So) and noise (No) at the two ears (NoSo) and to identical noise but with the signal inverted at one ear (NoS pi): the noise was 7-15 dB suprathreshold. The masked threshold was estimated by the standard separation, "D". The neural BMLD was estimated as the difference between the masked thresholds for NoSo and NoS pi. The presence of So and S pi tones was indicated by discharge rate increases in 55.3% of neurons. In 36.4% of neurons, the presence of So tones was indicated by an increase in discharge rate and S pi tones by a decrease. In 6.8% of neurons, both So and S pi tones caused a decrease in discharge rate. In only 1.5% of neurons was So indicated by a decrease and S pi by an increase in discharge rate. Responses to the binaural configurations were consistent with the neuron's interaural delay sensitivities; 34.4% of neurons showing increases in discharge rate to both So and S pi tones gave positive BMLDs > or = 3 dB (S pi tones were detected at lower levels than So), whereas 37.3% gave negative BMLDs > or = 3 dB. For neurons in which So signals caused an increase in the discharge rate and S pi a decrease, 72.7% gave positive BMLDs > or = 3 dB and only 4.5% gave negative BMLDs > or = 3 dB. The results suggest that the responses of single ICC neurons are consistent with the psychophysical BMLDs for NoSo versus NoS pi at 500 Hz, and with current binaural interaction models based on coincidence detection. The neurons likely to contribute to the psychophysical BMLD are those with BFs near 500 Hz, but detection of So and S pi tones may depend on different populations of neurons. PMID- 9212260 TI - Functional organization of the projection from area 2 to area 4gamma in the cat. AB - It has been shown that tetanic stimulation of area 2 of the somatosensory cortex produces long-term potentiation in neurons in area 4gamma, and this has been suggested as the basis of learning new motor skills. The purpose of this study was to further elucidate the characteristics of this projection by the use of evoked potentials in area 4gamma elicited by intracortical microstimulation of area 2. The experiments were carried out in cats and the following results were obtained. 1) In six animals, stimulation of a given site in area 2 elicited evoked potentials in a restricted region of area 4gamma, the size of which ranged from 1 to 1.5 mm2. These responses were labeled "localized responses." The evoked potentials were recorded from the superficial layers of the cortex, and were positive monophasic in shape. 2) In 16 animals, stimulation of a given site in area 2 elicited a focal response that was surrounded by smaller positive and monophasic potentials of <50% amplitude that spread broadly over area 4gamma. These responses were labeled "graded responses." 3) The sites that produced focal evoked potentials in area 4gamma extended along the direction of the radial fibers in area 2. These sites were defined as most effective segments (MESs). 4) The receptive fields of neurons along the MES in area 2 were similar to those of neurons recorded at the foci of the evoked potentials in area 4gamma. The results demonstrate that some of the projections from area 2 to area 4gamma are highly specific and that the somatosensory and motor areas that are connected by these specific projections receive functionally related peripheral input. These results are discussed in relation to possible mechanisms underlying motor learning. PMID- 9212261 TI - Electrophysiological analysis of dorsal root ganglion neurons pre- and post coexpression of green fluorescent protein and functional 5-HT3 receptor. AB - Aequorea green fluorescent protein (GFP) is an excellent marker to examine genetically altered live cells in whole animals or culture. Its potential use in identifying genetically modified neurons, however, has not been investigated extensively. To examine the usefulness, toxicity, and potential electrophyiological effects of GFP expression in neurons, we generated adenovirus containing the mGFP4 cDNA. One week after virus transfection of dorsal root ganglion neurons (DRG), 10% of postnatal DRG neurons appeared brightly fluorescent, labelling the soma and neurites. Temporal examination of these neurons demonstrated no toxicity to DRG neurons even after several weeks in culture with repeated daily epifluorescent exposure. Electrophysiological analysis and comparison of control and viral exposed (GFP- and GFP+) DRG neurons did not demonstrate any differences in whole cell resistance, resting potential, action potential (AP) threshold, AP duration, AP amplitude, or whole cell capacitance. To investigate the usefulness of GFP as a marker for identifying neurons genetically altered to express a novel neurotransmitter receptor, a second adenovirus construct was generated containing both GFP and serotonin type 3 (5-HT3) receptor cDNAs. Transfection of DRG neurons with this virus produced an inward current in the presence of serotonin only in DRG neurons that were GFP positive. It is concluded that adenoviral transfection of neurons with GFP, for cellular labeling, and coexpression of GFP-neurotransmitter constructs are safe, nontoxic, methods for electrophysiologically investigating neurons over several weeks. The uniqueness of the vector used in these experiments is that it was constructed to express GFP in a second cassette so that it would label the transduced cells, but have no potential for interfering with the function of the foreign 5-HT3 receptor. PMID- 9212262 TI - Recordings from brain stem neurons responding to chemical stimulation of the subarachnoid space. AB - The subarachnoid space at the base of the skull was perfused continuously with artificial cerebrospinal fluid in anesthetized rats. A combination of inflammatory mediators consisting of histamine, bradykinin, serotonin, and prostaglandin E2 (10(-5) M) at pH of 6.1 was introduced into the flow for defined periods to stimulate meningeal primary afferents. Secondary neurons in the caudal nucleus of the trigeminal brain stem were searched by electrical stimulation of the cornea. Of the units receiving oligosynaptic input from the cornea, 44% were excited by stimulation of the meninges with inflammatory mediators. Most of these units had small receptive fields including cornea and the periorbital region, and their responsiveness was restricted to stimuli of noxious intensity. Three types of responses to stimulation of the meninges with algogenic agents were encountered: responses that did not outlast the stimulus period, responses outlasting the stimulus period for several minutes, and oscillating response patterns containing periods of enhanced and suppressed activity. The response pattern of a unit was reproducible, however, upon repetitive stimulation at 20 min intervals; the response magnitude showed tachyphylaxis upon stimulus repetition. The preparation presented mimics pathophysiolocial states normally accompanied by headache, e.g., subarachnoidal bleeding. Responsiveness of neurons in the caudal nucleus of the trigeminal brain stem to inflammatory mediators may play a role in the generation and maintenance of headache, e.g., migraine. PMID- 9212263 TI - Recruitment of GABAergic inhibition and synchronization of inhibitory interneurons in rat neocortex. AB - Intracellular recordings were obtained from pyramidal and interneuronal cells in rat neocortical slices to examine the recruitment of GABAergic inhibition and inhibitory interneurons. In the presence of the convulsant agent 4-aminopyridine (4-AP), after excitatory amino acid (EAA) ionotropic transmission was blocked, large-amplitude triphasic inhibitory postsynaptic potentials (IPSPs) occurred rhythmically (every 10-40 s) and synchronously in pyramidal neurons. After exposure to the gamma-aminobutyric acid-A (GABA(A)) receptor antagonist picrotoxin, large-amplitude monophasic slow IPSPs persisted in these cells. In the presence of 4-AP and EAA blockers, interneurons showed periodic spike firing. Although some spikes rode on an underlying synaptic depolarization, much of the rhythmic firing consisted of spikes having highly variable amplitudes, arising abruptly from baseline, even during hyperpolarization. The spike firing and depolarizing synaptic potentials were completely suppressed by picrotoxin exposure, although monophasic slow IPSPs persisted in interneurons. This suggests that this subset of interneurons may participate in generating fast GABA(A) IPSPs, but not slow GABA(B) IPSPs. Cell morphology was confirmed by intracellular injection of neurobiotin or the fluorescent dye Lucifer yellow CH. Dye injection into interneurons often (>70%) resulted in the labeling of two to six cells (dye coupling). These findings suggest that GABA(A)ergic neurons may be synchronized via recurrent collaterals through the depolarizing action of synaptically activated GABA(A) receptors and a mechanism involving electrotonic coupling. Although inhibitory neurons mediating GABA(B) IPSPs may be entrained by the excitatory GABA(A) mechanism, they appear to be a separate subset of GABAergic neurons capable of functioning independently with autonomous pacing. PMID- 9212264 TI - Synchronized oscillations in the inferior olive are controlled by the hyperpolarization-activated cation current I(h). AB - The participation of a hyperpolarization-activated cationic current in the generation of oscillations in single inferior olive neurons and in the generation of ensemble oscillations in the inferior olive nucleus (IO) of the guinea pig and ferret was investigated in slices maintained in vitro. Intracellular recordings in guinea pig or ferret 10 neurons revealed that these cells could generate sustained endogenous oscillations (4-10 Hz) at hyperpolarized membrane potentials (-60 to -67 mV) after the intracellular injection of a brief hyperpolarizing current pulse. These oscillations appeared as the rhythmic generation of a low threshold Ca2+ spike that typically initiated one or two fast Na+-dependent action potentials. Between low-threshold Ca2+ spikes was an afterhyperpolarization that formed a "pacemaker" potential. Local application of apamin resulted in a large reduction in the amplitude of the afterhyperpolarization, indicating that a Ca2+-activated K+ current makes a strong contribution to its generation. However, even in the presence of apamin, hyperpolarization of IO neurons results in a "depolarizing sag" of the membrane potential that was blocked by local application of Cs+ or partial replacement of extracellular Na+ with choline+ or N-methyl-D-glucamine+, suggesting that I(h) also contributes to the generation of the afterhyperpolarization. Extracellular application of low concentrations of cesium resulted in hyperpolarization of the membrane potential of IO neurons and spontaneous 5- to 6-Hz oscillations in single, as well as networks, of IO neurons. Application of larger concentrations of cesium reduced the frequency of oscillation to 2-3 Hz or blocked the oscillation entirely. On the basis of these results, we propose that I(h) contributes to single and ensemble oscillations in the IO in two ways: 1) I(h) contributes to the determination of the resting membrane potential such that reduction of I(h) results in hyperpolarization of the membrane potential and an increased propensity of oscillation through removal of inactivation of the low threshold Ca2+ current; and 2) I(h) contributes to the generation of the afterhyperpolarization and the pacemaker potential between low-threshold Ca2+ spikes. PMID- 9212265 TI - Afferent inputs modulate the activity of a rhythmic burst generator in the rat disinhibited spinal cord in vitro. AB - Application of strychnine and bicuculline to the isolated spinal cord of the neonatal rat induces spontaneous bursting of regular rhythmicity (cycle period approximately 30 s). This phenomenon is important because it shows that a spinal network, made up by excitatory connections only, generates a very reliable rhythmic pattern. To find out how signals from the periphery or higher centres might influence the operation of the rhythmogenic network, the present experiments examined whether synaptic inputs from dorsal root (DR) or ventrolateral (VL) afferent fibers could modulate this spontaneous rhythmicity. This issue was addressed with intracellular recording from motoneurons or extracellular recording from ventral roots after eliciting bursting with strychnine plus bicuculline. Single electrical shocks (0.1 ms; intensity 1-4 times threshold) applied to one DR reset spontaneous bursting without altering its period or duration. Repetitive stimulations at periods ranging from 20 to 2 s entrained bursts on a 1:1 basis. Burst duration was shorter at lower stimulation periods whereas burst amplitude was unchanged. The lowest stimulation period compatible with burst entrainment depended on stimulus strength. At stimulation periods <2-s entrainment was always lost and spontaneous bursts unexpectedly returned even if electrical pulses still elicited ventral root reflexes. Such spontaneous bursts had similar properties as those recorded in the absence of electrical pulses. Analogous results were obtained with VL stimulations. It is concluded that the spinal rhythmogenic network was highly susceptible to external synaptic inputs, which paced burst generation whereas burst duration was adapted to interstimulus interval. A scheme is provided to explain the modulatory role of synaptic inputs as well as the escape of bursting from fast stimulus entrainment in terms of a rhythmogenic network functionally separated from reflex pathways activated by DR or VL tracts. PMID- 9212266 TI - Regulation of masticatory force during cortically induced rhythmic jaw movements in the anesthetized rabbit. AB - To examine the relationships between masticatory force, electromyogram (EMG) of masticatory muscles, and jaw movement pattern, we quantitatively evaluated the effects of changing hardness of a chewing substance on these three variables. Cortically induced rhythmic jaw movements of a crescent-shaped pattern were induced by electrical stimulation of the cerebral cortical masticatory area in the anesthetized rabbit. The axially directed masticatory force was recorded with a small force-displacement transducer mounted on the ground surface of the lower molars. EMGs were recorded from the masseter and digastric muscles simultaneously with jaw movements. Five test strips of polyurethane foam of different hardness were prepared and inserted between the upper molar and the transducer during the movements. The peak, impulse, and buildup speed of the masticatory force increased with strip hardness, whereas duration of the exerted force did not vary with strip hardness. The integrated activity and duration of the masseteric EMG bursts also increased with strip hardness. The integrated EMG activity of the digastric bursts was weakly related to strip hardness, whereas the duration was not. The minimum gape increased with strip hardness, but the maximum gape did not. The horizontal excursion of the jaw did not vary in a hardness-dependent manner, although it was greater in the cycles with strip application than in the cycles without strip application. Deprivation of periodontal sensation by cutting the nerves to the teeth reduced the buildup speed of the force, maximum gape, net gape, and horizontal jaw movements. The denervation also elongated the force duration and that of masseteric EMG bursts. However, the rate of the hardness dependent changes in the above parameters did not alter after denervation. The latency of the masseteric EMG response to strip application was evaluated before and after denervation. In both conditions, it was > or = 6 ms in approximately 70% of the cycles and <6 ms in the remaining approximately 30%, which cannot be explained by a simple reflex mechanism. On the basis of the analysis of correlation coefficients, the masseteric integrated EMG seemed to be a good indicator of the peak and impulse of the masticatory force both before and after denervation. We conclude that periodontal afferents would be responsible for a quick buildup of masticatory force and that afferents other than those from periodontal tissue would contribute to the hardness-dependent change of masticatory force during cortically induced rhythmic jaw movements. PMID- 9212267 TI - Spatial memory and path integration studied by self-driven passive linear displacement. I. Basic properties. AB - According to path integration, the brain is able to compute the distance of a traveled path. In this research we applied our previously reported method for studying memory of linear distance, a crucial mechanism in path integration; our method is based on the overt reconstruction of a passive transport. Passive transport is a special case of navigation in which no active control is performed. Blindfolded subjects were first asked to travel 2 m forward, in darkness, by driving with a joystick the robot on which they were seated. The results show that all subjects but two undershot this distance, i.e., overestimated their own displacement. Then, subjects were submitted to a passive linear forward displacement along 2, 4, 6, 8, or 10 m, and had to reproduce the same distance, still blindfolded. The results show that the distance of the stimulus was accurately reproduced, as well as stimulus duration, peak velocity, and velocity profile. In this first condition, the imposed velocity profile was triangular and therefore stimulus distance and duration were correlated. In a second condition, it was shown that distance was correctly reproduced also when the information about stimulus duration was kept constant. Here, different velocity profiles were used as stimuli, and most subjects also reproduced the velocity profile. Statistical analyses indicated that distance was not reproduced as a consequence of duration, peak velocity, or velocity profile reproduction, but was uniquely correlated to stimulus distance. The previous hypothesis of a double integration of the otolith signal to provide a distance estimate can explain our results. There was a large discrepancy between the accuracy with which the subjects matched the velocity profiles and that of distance reproduction. It follows that, whereas the dynamics of passive motion are stored and available to further use, distance is independently estimated. It is concluded that vestibular and somatosensory signals excited by passive transport can be used to build a dynamic as well as a static representation of the traveled path. We found a close quantitative similarity between the present findings on distance reproduction and those obtained from active locomotion experiments in which the same paradigm was used. This resemblance suggests that the two types of navigation tasks draw on common physiological processes and extends the relevance of our results to naturally occurring path integration. PMID- 9212268 TI - Visual field representation in striate and prestriate cortices of a prosimian primate (Galago garnetti). AB - Microelectrode mapping techniques were used to study the visuotopic organization of the first and second visual areas (V1 and V2, respectively) in anesthetized Galago garnetti, alorisiform prosimian primate. 1) V1 occupies approximately 200 mm2 of cortex, and is pear shaped, rather than elliptical as in simian primates. Neurons in V1 form a continuous (1st-order) representation of the visual field, with the vertical meridian forming most of its perimeter. The representation of the horizontal meridian divides V1 into nearly equal sectors representing the upper quadrant ventrally, and the lower quadrant dorsally. 2) The emphasis on representation of central vision is less marked in Galago than in simian primates, both diurnal and nocturnal. The decay of cortical magnification factor with increasing eccentricity is almost exactly counterbalanced by an increase in average receptive field size, such that a point anywhere in the visual field is represented by a compartment of similar diameter in V1. 3) Although most of the cortex surrounding V1 corresponds to V2, one-quarter of the perimeter of V1 is formed by agranular cortex within the rostral calcarine sulcus, including area prostriata. Although under our recording conditions virtually every recording site in V2 yielded visually responsive cells, only a minority of those in area prostriata revealed such responses. 4) V2 forms a cortical belt of variable width, being narrowest (approximately 1 mm) in the representation of the area centralis and widest (2.5-3 mm) in the representation of the midperiphery (>20 degrees eccentricity) of the visual field. V2 forms a second-order representation of the visual field, with the area centralis being represented laterally and the visual field periphery medially, near the calcarine sulcus. Unlike in simians, the line of field discontinuity in Galago V2 does not exactly coincide with the horizontal meridian: a portion of the lower quadrant immediately adjacent to the horizontal meridian is represented at the rostral border of ventral V2, instead of in dorsal V2. Despite the absence of cytochrome oxidase stripes, the visual field map in Galago V2 resembles the ones described in simians in that the magnification factor is anisotropic. 5) Receptive field progressions in cortex rostral to dorsal V2 suggest the presence of a homologue of the dorsomedial area, including representations of both quadrants of the visual field. These results indicate that many aspects of organization of V1 and V2 in simian primates are shared with lorisiform prosimians, and are therefore likely to have been present in the last common ancestor of living primates. However, some aspects of organization of the caudal visual areas in Galago are intermediate between nonprimates and simian primates, reflecting either an intermediate stage of differentiation or adaptations to a nocturnal niche. These include the shape and the small size of V1 and V2, the modest degree of emphasis on central visual field representation, and the relatively large area prostriata. PMID- 9212269 TI - Synaptic connectivity in cultured hypothalamic neuronal networks. AB - We have developed a novel approach to analyze the synaptic connectivity of spontaneously active networks of hypothalamic neurons in culture. Synaptic connections were identified by recording simultaneously from pairs of neurons using the whole cell configuration of the patch-clamp technique and testing for evoked postsynaptic current responses to electrical stimulation of one of the neurons. Excitatory and inhibitory responses were distinguished on the basis of their voltage and time dependence. The distribution of latencies between presynaptic stimulation and postsynaptic response showed multiple peaks at regular intervals, suggesting that responses via both monosynaptic and polysynaptic paths were recorded. The probability that an excitatory event is transmitted to another excitatory neuron and results in an above-threshold stimulation was found to be only one in three to four. This low value indicates that in addition to evoked synaptic responses other sources of excitatory drive must contribute to the spontaneous activity observed in these networks. The various types of synaptic connections (excitatory and inhibitory, monosynaptic, and polysynaptic) were counted, and the observations analyzed using a probabilistic model of the network structure. This analysis provides estimates for the ratio of inhibitory to excitatory neurons in the network (1:1.5) and for the ratio of postsynaptic cells receiving input from a single GABAergic or glutamatergic neuron (3:1). The total number of inhibitory synaptic connections was twice that of excitatory connections. Cell pairs mutually connected by an excitatory and an inhibitory synapse occurred significantly more often than predicted by a random process. These results suggests that the formation of neuronal networks in vitro is controlled by cellular mechanisms that favor inhibitory connections in general and specifically enhance the formation of reciprocal connections between pairs of excitatory and inhibitory neurons. These mechanisms may contribute to network formation and function in vivo. PMID- 9212270 TI - Positive force feedback control of muscles. AB - This study was prompted by recent evidence for the existence of positive force feedback in feline locomotor control. Our aim was to establish some basic properties of positive force feedback in relation to load compensation, stability, intrinsic muscle properties, and interaction with displacement feedback. In human subjects, muscles acting about the wrist and ankle were activated by feedback-controlled electrical stimulation. The feedback signals were obtained from sensors monitoring force and displacement. The signals were filtered to mimic transduction by mammalian tendon organ and muscle spindle receptors. We found that when muscles under positive force feedback were loaded inertially, they responded in a stable manner with increased active force. The activation attenuated the muscle stretch (yield) that would otherwise occur in the absence of feedback. With enough positive force feedback gain, yield could actually reverse. This behavior, which we termed the affirming reaction, was reminiscent of the mammalian positive supporting reaction, a postural response elicited by contact of the foot with the ground. Muscles under positive force feedback remained stable, even when the loop gain (Gf) was set at levels of 2 or 3. In a linear system, if Gf > 1, instability occurs when the loop is closed. On further investigation, we found that Gf changed with joint angle: it declined as the load-bearing muscle actively shortened. We inferred that in closed-loop operation, the active muscles always shortened until Gf approached unity. In other words, the length-tension curve of active muscle ensures stability even when force-related excitation of motoneurons is very large. Concomitant negative displacement feedback reinforced and stabilized load compensation up to a certain gain, beyond which instability occurred. In further trials we included delays of up to 40 ms in the positive force feedback pathway, to model the delays recently described for tendon organ reflexes in cat locomotion. Contrary to expectations, this did not destabilize the loop. Indeed, when instability was deliberately evoked by setting displacement feedback gain high, delays in the positive force feedback pathway actually stabilized control. The stabilization of positive force feedback by inherent properties of the neuromuscular system increases the functional scope to be expected of feedback from force receptors in biological motor control. Our results provide a rationale for the delayed excitatory action of Ib heteronymous input on extensor motoneurons in cat locomotion. PMID- 9212271 TI - Implications of positive feedback in the control of movement. AB - In this paper we review some theoretical aspects of positive feedback in the control of movement. The focus is mainly on new theories regarding the reflexive role of sensory signals from mammalian tendon organ afferents. In static postures these afferents generally mediate negative force feedback. But in locomotion there is evidence of a switch to positive force feedback action. Positive feedback is often associated with instability and oscillation, neither of which occur in normal locomotion. We address this paradox with the use of analytic models of the neuromuscular control system. It is shown that positive force feedback contributes to load compensation and is surprisingly stable because the length-tension properties of mammalian muscle provide automatic gain control. This mechanism can stabilize control even when positive feedback is very strong. The models also show how positive force feedback is stabilized by concomitant negative displacement feedback and, unexpectedly, by delays in the positive feedback pathway. Other examples of positive feedback in animal motor control systems are discussed, including the beta-fusimotor system, which mediates positive feedback of displacement. In general it is seen that positive feedback reduces the sensitivity of the controlled extremities to perturbations of posture and load. We conclude that positive force feedback can provide stable and effective load compensation that complements the action of negative displacement and velocity feedback. PMID- 9212272 TI - Quantitative analysis of orofacial thermoreceptive neurons in the superficial medullary dorsal horn of the rat. AB - Surprisingly little is known concerning the central processing of innocuous thermal somatosensory information. The aim of the present study was to obtain quantitative data on the characteristics of neurons in the rat superficial medullary dorsal horn (sMDH) that responded to innocuous thermal stimulation of the rat's face and tongue. Single-unit extracellular recordings were obtained in chloralose-urethane anesthetized rats. A total of 153 thermoreceptive neurons was studied. Of these, 146 were excited by cooling and inhibited by warming and were classified as COLD cells. The remaining seven cells were excited by innocuous warming of the skin or tongue. Of 123 COLD cells tested, 33% were excited by touch and 22% by pinch stimuli delivered to the thermoreceptive field. Of the 50 COLD cells tested, 46% were excited also by noxious heating (> or = 50 degrees C for 5 s). Most (82/121) of the receptive fields were located on the upper lip, 25 on the tongue, and most of the remaining on the lower lip. Receptive fields were generally small (1-5 mm2). In some experiments, electrical stimulation in the thalamus was performed, and nine COLD cells could be activated antidromically. The responses of 38 COLD cells to incremental 5 degrees C cooling steps were examined quantitatively. Thermal stimuli were applied to facial or lingual receptive fields of sMDH neurons with a computer-controlled Peltier thermode starting from 33 degrees C, decreasing to 8 or 3 degrees C, and returning to 33 degrees C. Most COLD cells (26/38) had both static and dynamic responses; 7 had mainly dynamic and 5 mainly static responses to step decreases in temperature. Rat sMDH COLD cells could be classified into three groups depending on their stimulus-response functions. The first group (Type 1, n = 19) had a bell-shaped static stimulus response function. The second group (Type 2) had a high maintained or increasing static firing rate as the temperature decreased < 18 degrees C (n = 10). Type 3 COLD cells had mainly dynamic properties (n = 7). Many of the cells in all groups were excited by noxious mechanical stimulation. Type 2 cells differed from the other two groups in that most did not respond to noxious thermal stimuli (hot) and many responded to innocuous tactile stimuli. Neurons from each of the three groups of COLD cells could be activated antidromically from contralateral thalamus. These data suggest that there is little central processing of thermal information at the first central synapse for Type 1 neurons, however, the responses of the other two types may be due to central processing and convergence. The demonstration of rat sMDH COLD cells with distinctive stimulus-response functions to thermal shifts suggests separate functional roles of these neurons in the ascending thermal sensory pathway. PMID- 9212273 TI - Invariant visual responses from attentional gain fields. AB - Inferotemporal (IT) neurons exhibit a substantial degree of invariance with respect to translation of images used as visual stimuli. Through theoretical and computer-modeling methods, we show how translation-invariant receptive fields, like those of IT neurons, can be generated from the responses of V4 neurons if the effects of attention are taken into account. The model incorporates a recently reported form of attention-dependent gain modulation in V4 and produces IT receptive fields that shift so they are centered at the point where attention is directed. Receptive fields of variable, attention-controlled spatial scale are obtained when the mechanism is extended to scale-dependent attentional gain fields. The results indicate that gain modulation may play analogous roles in the dorsal and ventral visual pathways, generating transformations from retinal coordinates to body- and object-centered systems, respectively. PMID- 9212274 TI - Role of calcium conductances on spike afterpotentials in rat trigeminal motoneurons. AB - Intracellular recordings were obtained from rat trigeminal motoneurons in slice preparations to investigate the role of calcium conductances in the depolarizing and hyperpolarizing spike afterpotential (ADP and mAHP, respectively). The mAHP was suppressed by bath application of 1 microM apamin, 2 mM Mn2+, and 2 mM Co2+, and also by intracellular injection of ethylene glycol-bis(b-aminoethylenether) N,N,N',N'-tetraacetic acid (EGTA), suggesting that the potassium conductance generating the mAHP is activated by Ca2+ influx. Mn2+ (2 mM) or Cd2+ (500 microM) reduced the ADP, whereas the ADP amplitude was increased by raising extracellular Ca2+ concentration from 2 to 8 mM by bath application of Ba2+ (0.5-5 mM) and by intracellular injection of EGTA. This would suggest that Ca2+ itself is likely to be the charge carrier generating the ADP. Focal application of omega-conotoxin GVIA (10-30 microM) suppressed the mAHP and enhanced the ADP, whereas focal application of omega-agatoxin IVA (10-100 microM) reduced the ADP amplitude without apparent effects on the mAHP. We conclude that Ca2+ influx through omega agatoxin IVA-sensitive calcium channels is at least in part responsible for the generation of the ADP and that Ca2+ influx through omega-conotoxin GVIA-sensitive calcium channels contributes to the generation of the mAHP. Because of the selective suppression of the ADP and mAHP by omega-agatoxin IVA and omega conotoxin GVIA, respectively, it is suggested that both calcium channels are separated geometrically in rat trigeminal motoneurons. PMID- 9212275 TI - Two parallel signaling pathways couple 5HT1A receptors to N- and L-type calcium channels in C-like rat dorsal root ganglion cells. AB - The coupling of serotonin receptors to Ca2+ channels was studied in a subpopulation of acutely isolated rat dorsal root ganglion (DRG) cell bodies (type 1 DRG cells), which have membrane properties similar to C-type nociceptive sensory neurons. In these cells, serotonin (5HT) inhibited high-threshold Ca2+ channel current and decreased action potential duration. The inhibitory effects of 5HT and the 5HT1A agonist 8-OH-DPAT were shown to be antagonized by the 5HT1A antagonists spiperone and pindolol, respectively, indicating involvement of a 5HT1A receptor. Several observations suggest that 5HT1A receptors couple to N- and L-type Ca2+ channels by two different signaling pathways in type 1 DRG cells. The inhibition of Ca2+ channel currents produced by 10 microM 5HT occurred in two phases, an initial slowing of current activation rate (kinetic slowing), which was complete within 10 s, and a simultaneous reduction in steady state current amplitude (steady state inhibition), which peaked in approximately 1 min. The kinetic slowing, but not steady state inhibition, was reversed by a positive prepulse to +70 mV (prepulse). Blockade of N-type Ca2+ channels selectively reduced the kinetic slowing and its reversal by prepulses. Chelation of intracellular Ca2+ or blockade of L-type Ca2+ channels selectively reduced the steady state inhibition. Recordings using the cell-attached patch configuration suggest that steady state inhibition required a component that was diffusible in the cytosol, while kinetic slowing occurred via a membrane delimited pathway. The application of 5HT to the cell body outside the patch pipette reduced macroscopic Ca2+ channel currents in 33% of small-diameter DRG cells tested, indicating the participation of a cytosolic diffusible component. Application of 5HT (a membrane impermeant compound) outside the patch pipette produced steady state inhibition only, whereas similar application of membrane permeant 5HT1A agonists, 8-OH-DPAT or 5-methoxy-N,N-dimethyl-tryptamine, produced kinetic slowing and steady state inhibition. Together these data suggest that 5HT1A receptors couple negatively to Ca2+ channels via two pathways: a membrane-delimited pathway that couples to N channels and actuates voltage-sensitive kinetic slowing and a pathway dependent on a cytosolic diffusible component and free intracellular Ca2+, which couples to L channels and actuates steady state inhibition. PMID- 9212276 TI - Dynamics of neurons controlling movements of a locust hind leg. III. Extensor tibiae motor neurons. AB - Imposed movements of the apodeme of the femoral chordotonal organ (FeCO) of the locust hind leg elicit resistance reflexes in extensor and flexor tibiae motor neurons. The synaptic responses of the fast and slow extensor tibiae motor neurons (FETi and SETi, respectively) and the spike responses of SETi were analyzed with the use of the Wiener kernel white noise method to determine their response properties. The first-order Wiener kernels computed from soma recordings were essentially monophasic, or low passed, indicating that the motor neurons were primarily sensitive to the position of the tibia about the femorotibial joint. The responses of both extensor motor neurons had large nonlinear components. The second-order kernels of the synaptic responses of FETi and SETi had large on-diagonal peaks with two small off-diagonal valleys. That of SETi had an additional elongated valley on the diagonal, which was accompanied by two off diagonal depolarizing peaks at a cutoff frequency of 58 Hz. These second-order components represent a half-wave rectification of the position-sensitive depolarizing response in FETi and SETi, and a delayed inhibitory input to SETi, indicating that both motor neurons were directionally sensitive. Model predictions of the responses of the motor neurons showed that the first-order (linear) characterization poorly predicted the actual responses of FETi and SETi to FeCO stimulation, whereas the addition of the second-order (nonlinear) term markedly improved the performance of the model. Simultaneous recordings from the soma and a neuropilar process of FETi showed that its synaptic responses to FeCO stimulation were phase delayed by about -30 degrees at 20 Hz, and reduced in amplitude by 30-40% when recorded in the soma. Similar configurations of the first and second-order kernels indicated that the primary process of FETi acted as a low-pass filter. Cross-correlation between a white noise stimulus and a unitized spike discharge of SETi again produced well-defined first- and second order kernels that showed that the SETi spike response was also dependent on positional inputs. An elongated negative valley on the diagonal, characteristic of the second-order kernel of the synaptic response in SETi, was absent in the kernel from the spike component, suggesting that information is lost in the spike production process. The functional significance of these results is discussed in relation to the behavior of the locust. PMID- 9212277 TI - Cutaneous reflexes during human gait: electromyographic and kinematic responses to electrical stimulation. AB - The functions of ipsilateral cutaneous reflexes were studied with short trains of stimuli presented pseudorandomly to the superficial peroneal (SP) and tibial nerves during human gait. Electromyograms (EMGs) of tibialis anterior (TA), soleus, lateral and medial gastrocnemius, vastus lateralis (VL), and biceps femoris (BF) muscle were recorded, together with ankle and knee joint angles. Net reflex EMG responses were quantified in each of the 16 parts of the step cycle according to a recently developed technique. After SP nerve stimulation, TA muscle showed a significant suppression during swing phase that was highly correlated to ankle plantarflexion. BF and VL muscles were both excited throughout swing and significantly correlated to knee flexion during early swing. Tibial nerve stimulation caused dorsiflexion during late stance, but plantarflexion during late swing. We argue that SP nerve reflexes are indicative of a stumbling corrective response to nonnoxious electrical stimulation in humans. The correlated kinematic responses after tibial nerve stimulation may allow smooth movement of the swing leg so as to prevent tripping during swing and to assist placing and weight acceptance at the beginning of stance. PMID- 9212278 TI - Noradrenergic suppression of synaptic transmission may influence cortical signal to-noise ratio. AB - Norepinephrine has been proposed to influence signal-to-noise ratio within cortical structures, but the exact cellular mechanisms underlying this influence have not been described in detail. Here we present data on a cellular effect of norepinephrine that could contribute to the influence on signal-to-noise ratio. In brain slice preparations of the rat piriform (olfactory) cortex, perfusion of norepinephrine causes a dose-dependent suppression of excitatory synaptic potentials in the layer containing synapses among pyramidal cells in the cortex (layer Ib), while having a weaker effect on synaptic potentials in the afferent fiber layer (layer Ia). Effects of norepinephrine were similar in dose-response characteristics and laminar selectivity to the effects of the cholinergic agonist carbachol, and combined perfusion of both agonists caused effects similar to an equivalent concentration of a single agonist. In a computational model of the piriform cortex, we have analyzed the effect of noradrenergic suppression of synaptic transmission on signal-to-noise ratio. The selective suppression of excitatory intrinsic connectivity decreases the background activity of modeled neurons relative to the activity of neurons receiving direct afferent input. This can be interpreted as an increase in signal-to-noise ratio, but the term noise does not accurately characterize activity dependent on the intrinsic spread of excitation, which would more accurately be described as interpretation or retrieval. Increases in levels of norepinephrine mediated by locus coeruleus activity appear to enhance the influence of extrinsic input on cortical representations, allowing a pulse of norepinephrine in an arousing context to mediate formation of memories with a strong influence of environmental variables. PMID- 9212279 TI - Primary afferent depolarizations of sensory origin within contact-sensitive mechanoreceptive afferents of a crayfish leg. AB - Recordings from the central branches of single identified dactyl sensory afferent (DSA) neurons in a crayfish in vitro preparation were performed to study modifications of the sensory message occurring before the first central synapse. These afferents comprised hairs and force-sensitive mechanoreceptors with phasic and phasotonic response characteristics in the terminal segment (dactyl) of the crayfish leg. More than one afferent spike size was often observed in intracellular recordings from these afferents, thus indicating the presence of electrical coupling between the central processes of DSA fibers. Additionally, in identified DSA fibers with large spike sizes, primary afferent depolarizations (PADs) of up to 15 mV were observed, which sometimes triggered antidromic spikes in the afferent. Nevertheless, PADs were clearly inhibitory, because they shunted the afferent spikes. They exhibited the following properties. First, each PAD was preceded by an afferent spike from a neighboring hair, indicating that the PADs had a sensory rather than central origin. Second, PADs could follow high frequencies of afferent discharges without failure, a property suggestive of monosynaptic connections, but because PAD latencies varied by +/-0.5 ms it is more likely that they were mediated by a disynaptic pathway. Third, although PADs were evoked in an extremely reliable manner, their amplitude varied in a quantal manner. Most unitary PADs were the result of the release of < 12 quanta, the mean quantal content lying between 4 and 5; quantal size was large, approximately 1 mV. Fourth, PADs showed facilitation in some fibers, whereas in others they became much smaller when occurring at brief intervals. We suggest that PADs may be an efficient and parsimonious way to limit sensory inflow in space and time, allowing the crayfish to identify precisely both weak and strong mechanical stimuli. PMID- 9212280 TI - Glutamate currents in morphologically identified human dentate granule cells in temporal lobe epilepsy. AB - Glutamate-receptor-mediated synaptic transmission was studied in morphologically identified hippocampal dentate granule cells (DGCs; n = 31) with the use of whole cell patch-clamp recording and intracellular injection of biocytin or Lucifer yellow in slices prepared from surgically removed medial temporal lobe specimens of epileptic patients (14 specimens from 14 patients). In the current-clamp recording, low-frequency stimulation of the perforant path generated depolarizing postsynaptic potentials that consisted of excitatory postsynaptic potentials and phase-inverted inhibitory postsynaptic potentials mediated by the gamma aminobutyric acid-A (GABA(A)) receptor at a resting membrane potential of -62.7 +/- 2.0 (SE) mV. In the voltage-clamp recording, two glutamate conductances, a fast alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA)-receptor mediated excitatory postsynaptic current (EPSC; AMPA EPSC) and a slowly developing N-methyl-D-aspartate (NMDA)-receptor-mediated EPSC (NMDA EPSC), were isolated in the presence of a GABA(A) receptor antagonist. NMDA EPSCs showed a voltage-dependent increase in conductance with depolarization by exhibiting an N shaped current-voltage relationship. The slope conductance of the NMDA EPSC ranged from 1.1 to 9.4 nS in 31 DGCs, reaching up to twice the size of the AMPA conductance. This widely varying size of the NMDA conductance resulted in the generation of double-peaked EPSCs and a nonlinear increase of the slope conductance of up to 37.5 nS with positive membrane potentials, which resembled "paroxysmal currents," in a subpopulation of the neurons. In contrast, AMPA EPSCs, which were isolated in the presence of an NMDA receptor antagonist (2 amino-5-phosphonovaleric acid), showed voltage-independent linear changes in the current-voltage relationship and were blocked by 6-cyano-7-nitroquinoxaline-2,3 dione. The AMPA conductance showed little variance, regardless of the size of the NMDA conductance of a given neuron. The average AMPA slope conductance was 5.28 +/- 0.65 (SE) nS in 31 human DGCs. This value was similar to AMPA EPSC conductances in normal rat DGCs (5.35 +/- 0.52 nS, mean +/- SE; n = 55). Dendritic morphology and spine density were quantified in the individual DGCs to assess epileptic pathology. Dendritic spine density showed an inverse correlation (r2 = 0.705) with a slower rise time and a longer half-width of the excitatory postsynaptic potentials mediated by the NMDA receptor. It is concluded that both AMPA and NMDA EPSCs contribute to human DGC synaptic transmission in epileptic hippocampus. However, a wide range of changes in the slope conductance of the NMDA EPSCs suggests that the NMDA-receptor-mediated conductance could be altered in human epileptic DGCs. These changes may influence the generation of chronic subthreshold epileptogenic synaptic activity and give rise to pathological excitation leading to epileptic seizures and dendritic pathology. PMID- 9212281 TI - Functional MRI of pain- and attention-related activations in the human cingulate cortex. AB - The aims of the study were to use functional magnetic resonance imaging (fMRI) to 1) locate pain-related regions in the anterior cingulate cortex (ACC) of normal human subjects and 2) determine whether each subject's pain-related activation is congruent with ACC regions involved in attention-demanding cognitive processes. Ten normal subjects underwent fMRI with a 1.5-T standard commercial MRI scanner. A conventional gradient echo technique was used to obtain data from a single 4-mm sagittal slice of the left ACC, approximately 3.5 mm from midline. For each subject, interleaved sets of 6 images were obtained during a pain task, an attention-demanding task, and at rest, for a total of 36 images per task. Pain of different intensities was evoked via electrical stimulation of the right median nerve. The attention-demanding task consisted of silent word generation (verbal fluency). Additional experiments obtained data from the right ACC. A pixel-by pixel statistical analysis of task versus rest images was used to determine task related activated regions. The pain task resulted in a 1.6-4.0% increase in mean signal intensity within a small region of the ACC. The exact location of this activation varied from subject to subject, but was typically in the posterior part of area 24. The signal intensity changes within this region correlated with pain intensity reported by the subject. The attention-demanding tasks increased the mean signal intensity by 1.3-3.3% in a region anterior and/or superior to the pain-related activation in each subject. The activated region was typically larger than the pain-related activation. In some cases this activation was at or superior to the ACC border, near the supplementary motor area. These regions did not show any pain-intensity-related activation. In one subject both right and left ACC were imaged, revealing bilateral ACC activation during the attention task but only contralateral pain-related activation. These findings shed light on pain- and attention-related cognitive processes. The results provide evidence for a region in the posterior part of the ACC that is involved in pain and a more anterior region involved in other attention-demanding cognitive tasks. PMID- 9212282 TI - Ocular dominance peaks at pinwheel center singularities of the orientation map in cat visual cortex. AB - In the primary visual cortex of monkey and cat, ocular dominance and orientation are represented continuously and simultaneously, so that most neighboring neurons respond optimally to visual stimulation of the same eye and orientation. Maps of stimulus orientation are punctuated by singularities referred to as "pinwheel centers," around which all orientations are represented. Given that the orientation map is mostly continuous, orientation singularities are a mathematical necessity unless the map consists of perfectly parallel rows, and there is no evidence that the singularities play a role in normal function or development. We report here that in cats there is a strong tendency for peaks of ocular dominance to lie on the pinwheel center singularities of the orientation map. This relationship predicts but is not predicted by the tendencies, previously reported, for pinwheels to lie near the center lines of ocular dominance bands and for iso-orientation bands to cross ocular dominance boundaries at right angles. The coincidence of ocular dominance peaks with orientation singularities is likely to reflect a strong underlying functional link between the two visual cortical maps. PMID- 9212283 TI - Dissociation of saccade-related and pursuit-related activation in human frontal eye fields as revealed by fMRI. AB - The location of the human frontal eye fields (FEFs) underlying horizontal visually guided saccadic and pursuit eye movements was investigated with the use of functional magnetic resonance imaging in five healthy humans. Execution of both saccadic and pursuit eye movements induced bilateral FEF activation located medially at the junction of the precentral sulcus and the superior frontal sulcus and extending laterally to the precentral gyrus. These findings extend previous functional imaging studies by providing the first functional imaging evidence of a specific activation in the FEF during smooth pursuit eye movements in healthy humans. FEF activation during smooth pursuit performance was smaller than during saccades. This finding, which may reflect the presence of a smaller pursuit related region area in human FEF than the saccade-related region, is consistent with their relative size observed in the monkey. The mean location of the pursuit related FEF was more inferior and lateral than the location of the saccade related FEF. These results provide the first evidence that there are different subregions in the human FEF that are involved in the execution of two different types of eye movements, namely saccadic and pursuit eye movements. Moreover, this study provides additional evidence that the human FEF is located in Brodmann's area 6, unlike the monkey FEF which is located in the posterior part of Brodmann's area 8. PMID- 9212284 TI - Mg2+ inhibition of ATP-activated current in rat nodose ganglion neurons: evidence that Mg2+ decreases the agonist affinity of the receptor. AB - The effect of Mg2+ on ATP-activated current in rat nodose ganglion neurons was investigated with the use of the whole cell patch-clamp technique. Mg2+ decreased the amplitude of ATP-activated current in a concentration-dependent manner over the concentration range of 0.25-8 mM, with a 50% inhibitory concentration value of 1.5 mM for current activated by 10 microM ATP. Mg2+ shifted the ATP concentration-response curve to the right in a parallel manner, increasing the 50% effective concentration value for ATP from 9.2 microM in the absence of added Mg2+ to 25 microM in the presence of 1 mM Mg2+. Mg2+ increased the deactivation rate of ATP-activated current without changing its activation rate. The observations are consistent with an action of Mg2+ to inhibit ATP-gated ion channel function by decreasing the affinity of the agonist binding site on these receptors. PMID- 9212286 TI - Cortical control of human motoneuron firing during isometric contraction. AB - We recorded whole scalp magnetoencephalographic (MEG) signals simultaneously with the surface electromyogram from upper and lower limb muscles of six healthy right handed adults during voluntary isometric contraction. The 15- to 33-Hz MEG signals, originating from the anterior bank of the central sulcus, i.e., the primary motor cortex, were coherent with motor unit firing in all subjects and for all muscles. The coherent cortical rhythms originated in the hand motor area for upper limb muscles (1st dorsal interosseus, extensor indicis proprius, and biceps brachii) and close to the foot area for lower limb muscles (flexor hallucis brevis). The sites of origin corresponding to different upper limb muscles did not differ significantly. The cortical signals preceded motor unit firing by 12-53 ms. The lags were shortest for the biceps brachii and increased systematically with increasing corticomuscular distance. We suggest that the motor cortex drives the spinal motoneuronal pool during sustained contractions, with the observed cortical rhythmic activity influencing the timing of efferent commands. The cortical rhythms could be related to motor binding, but the rhythmic output may also serve to optimize motor cortex output during isometric contractions. PMID- 9212285 TI - Physiological evidence for local excitatory synaptic circuits in the rat hypothalamus. AB - We conducted whole cell voltage-clamp and current-clamp recordings in slices of rat hypothalamus to test for local excitatory synaptic circuits. Local excitatory inputs to neurons of the paraventricular nucleus (PVN) and supraoptic nucleus (SON) were studied with the use of electrical and chemical stimulation. Extracellular electrical stimulation provided indirect evidence of local excitatory circuits. Single stimuli evoked multiple excitatory postsynaptic potentials (EPSPs) or excitatory postsynaptic currents (EPSCs) in some PVN and SON cells, invoking polysynaptic excitatory inputs. Repetitive stimulation (10-20 Hz, 2-10 s) elicited long afterdischarges of EPSPs/EPSCs, suggesting a potentiation of upstream synapses in a polysynaptic circuit. Bath application of metabotropic glutamate receptor agonists provided more conclusive evidence for local excitatory circuits. Metabotropic receptor activation caused an increase in the frequency of EPSPs/EPSCs that was blocked by tetrodotoxin, suggesting that it was mediated by activation of local presynaptic excitatory neurons. The local excitatory inputs to SON and PVN neurons were mediated by glutamate release, because the EPSPs/EPSCs elicited with electrical stimulation and metabotropic receptor activation were blocked by ionotropic glutamate receptor antagonists. Finally, glutamate microstimulation furnished the most direct demonstration of local excitatory synaptic circuits. Glutamate microstimulation of perinuclear sites elicited an increase in the frequency of EPSPs/EPSCs in 13% of the PVN and SON neurons tested. Two sites provided most of the local excitatory synaptic inputs to PVN neurons, the dorsomedial hypothalamus and the perifornical region. These experiments provide converging physiological evidence for local excitatory synaptic inputs to hypothalamic neurons, inputs that may play a role in pulsatile hormone release. PMID- 9212287 TI - Human thalamic nucleus mediating taste and multiple other sensations related to ingestive behavior. AB - Until now, taste was the only primary sensory modality for which the human central nervous system pathways were unknown. We report sensations evoked by stimulation at microampere current levels in the region of the human thalamic nucleus (ventralis caudalis parvocellularis internis) corresponding to the monkey taste relay nucleus. Stimulation in this region during awake neurosurgical procedures evoked special visceral/somatic (taste/pungent smell), general visceral (fullness of a hollow viscus), as well as painful and nonpainful general somatic sensations. General somatic or visceral sensation was evoked by stimulation at 80% of sites where special visceral/somatic sensation was evoked. These results suggest that primate taste relay mediates multiple sensations in addition to taste. PMID- 9212288 TI - Marginal bone level after Le Fort I osteotomy. AB - The object of the study was to assess the effect of Le Fort I osteotomy and maxillary interdental osteotomy on the marginal bone level. Forty patients (25 female, 15 male, mean age 24 years, range 15-46) treated for dentofacial deformities comprised the subjects of the study and underwent Le Fort I osteotomy with or without simultaneous interdental osteotomy. Outcome was measured by marginal bone level measured in radiographs before and 1 year after operation. All patients had good oral hygiene. There was an overall significant mean marginal bone loss of 0.2 mm at surfaces without interdental osteotomy (P = 0.001). When the bone loss of the different types of teeth was considered separately, only those of central incisors (0.5 mm, P = 0.0001) and canines (0.4 mm, P = 0.004) were significant. Interdental osteotomy caused an overall mean marginal bone loss of 0.4 mm, but this was not significantly different from that of teeth without interdental osteotomy (P = 0.07). When the bone loss of different types of teeth after interdental osteotomy was considered separately, the only difference that achieved significance was that of premolars (0.3 mm, P = 0.04). Though there were significant differences, none of them was large enough to have any clinical relevance. Le Fort I osteotomy and interdental osteotomy may only in a few instances cause marginal bone loss of clinical relevance. However, the present study was performed on patients with good oral hygiene. The above conclusions may therefore not be valid for patients who, prior to surgery, already have a compromised marginal bone level. PMID- 9212289 TI - Comparison of different materials for interposition arthroplasty in treatment of temporomandibular joint ankylosis surgery: long-term follow-up in 25 cases. AB - A variety of interposition materials have been used to prevent recurrence after arthroplasty in treatment of temporomandibular joint ankylosis. The purpose of this retrospective study of our experience was to compare the different materials (skin, temporal muscle, homologous cartilage) used for interposition arthroplasty over a period of 22 years. A total of 25 patients (32 joints) with at least 3 years of follow-up were included. Good results were achieved in 92% of cases using total full thickness skin graft and 83% of cases using temporal muscle flap. Homologous cartilage gave poor results. PMID- 9212291 TI - Repair of experimental mandibular defects in rats with autogenous, demineralised, frozen and fresh bone. AB - This prospective experimental study aimed to assess the regenerative capability of demineralised bone autografts resected and replaced orthotopically, compared with traditional fresh and deep frozen mandibular autografts in rats. In 60 adult Wistar rats, a bone defect 4 x 4 mm was created at the left ascending mandibular ramus and the removed bone was used as a fresh (n = 20), deep frozen (n = 20), or demineralised (n = 20) graft which was implanted orthotopically 2 weeks later. Ten rats in each group were killed at 2 and 6 weeks later. Outcome was measured by cellular proliferation on histological examination. The number of mesenchymal cells was significantly greater (P < 0.05) at both 2 and 6 weeks in the demineralised grafts than in the other two groups. There were no differences between the 2- and 6-week examinations of deep frozen bone, nor between the medullary and peripheral aspects. It was concluded that demineralised bony autografts cause greater osteoinduction both in the short (2 weeks) and the medium (6 weeks) term. PMID- 9212290 TI - The role of oxygen free radicals in idiopathic facial pain. AB - Patients with chronic facial pain including those with facial arthromyalgia (TMJ dysfunction syndrome) were investigated for evidence of abnormal systemic and intra-articular free radical activity. Chronic facial pain patients showed significantly raised serum 2,3-dihydroxybenzoic acid after an oral dose of 1.2 g of aspirin which indicates increased systemic free radical activity. This was reflected in the TMJ aspirates of the facial arthromyalgia patients which contained thiobarbituric acid-reactive substance (TBA-RS) which is also a product of free radical activity. The synovial aspirates also contained high levels of the hyperalgesic eicosanoid 15-HETE. However, there was no difference between the painful and symptom-free joints, which suggested that in part the clinical features are probably determined by asymmetrical masticatory function or as yet unknown algesic factors such as local cytokine production. PMID- 9212292 TI - Protection of the lingual nerve during operations on the mandibular third molar: a simple method. AB - The object of the study was to assess the incidence of lingual nerve sensory loss during removal of impacted mandibular third molar teeth, and the effect of retention of the lingual plate on the incidence. The subjects were 395 patients, of whom 362 completed the study. Removal of 504 impacted wisdom teeth with retention of the lingual plate was performed. Results showed that 381 (76%) of the teeth were partially erupted and the remaining 123 were unerupted. In 497 (99%) bone had to be removed, and of these 376 (76%) required division of the tooth before removal (75% of the entire series). The only complication was transient paraesthesia in one patient which settled within a month. The study concludes that retention of the lingual plate gives optimum protection to the lingual nerve during removal of impacted wisdom teeth. PMID- 9212293 TI - Are the pharmacokinetics of ibuprofen important determinants for the drug's efficacy in postoperative pain after third molar surgery? AB - The aim of the present placebo-controlled, crossover study was to evaluate the efficacy and pharmacokinetics of ibuprofen after a single oral dose of soluble ibuprofen 400 mg in 18 patients with postoperative pain after bilateral third molar surgery. Throughout a 5-hour investigation period, patients reported significantly less pain (P < 0.001) after treatment with soluble ibuprofen than after placebo. Peak plasma concentrations of ibuprofen occurred approximately 30 minutes after dosage. No significant correlations (P > 0.05) were observed between efficacy measures and the various pharmacokinetic parameters (AUC0-300, Cmax and Tmax) for ibuprofen after the soluble dose. It is concluded that a single dose of soluble ibuprofen 400 mg is an effective analgesic for the control of postoperative pain in the early period after third molar surgery. Efficacy of this preparation does not appear to be directly related to the drug's pharmacokinetic variables in plasma. PMID- 9212294 TI - Micro-anatomy of primary lymphatics in oral mucosa: a scanning electron microscopic study. AB - Primary or initial lymphatics cannot readily be recognised by conventional histological methods because they have usually collapsed in excised tissues. We have investigated the distribution of 81 initial lymphatics in 24 specimens of normal tissue and specimens from 11 squamous cell carcinomas and their relationship with the invasion of carcinoma. We used indirect injection methods and the distended initial lymphatics were observed by scanning electron microscopy. The lymphatics were arranged in a network consisting of integrated lymph capillaries and precollectors in the subepithelial area. There was a zone of high density 100 microm below the epithelium. The arrangement of initial lymphatics in oral mucosa was similar to that in the skin of head, in contrast to initial lymphatics in the skin at other sites. We saw no carcinoma cells penetrating into initial lymphatics and the route of tumourous infiltration into lymphatics deserves further investigation. PMID- 9212295 TI - Granular-cell tumours: an immunohistochemical study. AB - The granular cell tumour (myoblastoma, Abrikosoff's tumour) and the congenital epulis in newborns (Neumann tumour) are two lesions rarely found in the oral cavity, whose histogenetic origin is highly controversial. This work analyses using immunohistochemical techniques 15 cases of myoblastomas and two of congenital epulis with different mono- and poly-clonal antibodies. Positive immunostaining was found for S-100 protein and neuron-specific enolase in all the cases of myoblastoma, and for vimentin and carcinoembryonic antigen in some cases. No immunoreactivity was observed for any of the other 13 antibodies used in congenital epulis. PMID- 9212296 TI - Oral pyogenic granuloma: a review of 38 cases from Ibadan, Nigeria. AB - Thirty-eight cases of patients with histologically confirmed pyogenic granuloma of the oral cavity were treated at the University College Hospital Dental Centre during the period 1982-1993. The age of the patients at presentation ranged from 5 to 74 years (mean 33) and the male:female ratio was 1:1.2. The main site was the gingiva (n-29, 74%). The clinical presentation was generally similar to that in other studies except that most of the lesions were large. All 38 cases were treated by excision and there were no recurrences among those with adequate follow-up. PMID- 9212297 TI - Osteomyelitis of the mandible in sickle cell disease. AB - We report 16 cases of osteomyelitis of the mandible in patients with sickle cell disease, an incidence of 5% of all patients who presented with osteomyelitis of the jaws. There was a striking predominance of men (13/16, 81%). The mean age at presentation was 23 years. The pathogens were predominantly mixed organisms and Staphylococcus aureus. Lesions were mainly in the posterior region of the mandible, and were usually operated on under general anaesthesia with adequate provision for the prevention of hypoxia. PMID- 9212298 TI - Cutaneous CD30 positive large T cell lymphoma of the upper lip: a rare presentation. AB - Primary cutaneous CD30 positive large T cell lymphomas (PCLCL) are very rare in the head and neck region. We report a case which presented on the right upper lip in a 48-year-old male. This was an isolated cutaneous lesion and in these circumstances this otherwise aggressive lymphoma has an excellent prognosis. PMID- 9212299 TI - Calcifying odontogenic cyst--a characteristic CT finding. AB - A report of two cases of calcifying odontogenic cyst in which the characteristic pathological finding of a peripheral band of intracystic calcification was demonstrated as a radiological sign on CT. PMID- 9212300 TI - Two new retractors for use in cryotherapy. AB - Two new instruments are described for use in the cryotherapy of peripheral nerves. They are designed to give good vision and access, protect surrounding tissues and be comfortable to hold. PMID- 9212301 TI - Carcinoma of the colon with mandible and liver metastases. PMID- 9212302 TI - Auriculotemporal syndrome with canities. PMID- 9212303 TI - Fibrosis in galeo pericranial flaps. PMID- 9212304 TI - Upper gastrointestinal bleeding following major maxillofacial surgery. PMID- 9212305 TI - Cognitive assessment in diabetes: the need for consensus. PMID- 9212306 TI - Vascular homeostasis, adhesion molecules, and macrovascular disease in non insulin-dependent diabetes mellitus. AB - Diabetes mellitus is characterized by fasting hyperglycaemia and the development of chronic vascular complications. While microvascular disease has been strongly related to glycaemic control, the major cause of mortality in diabetes is due to macrovascular disease affecting the cardiac and cerebrovascular circulations, which appear to have a more complex pathogenesis. Diabetes is associated with a 3 5-fold increase in death from myocardial infarction and similar figures pertain to stroke. The processes involved in atherothrombotic disease are complex and include variation in lipid metabolism, vascular responses, cell/cell interactions, and in the fluid and cellular phases of coagulation and fibrinolysis. The complex interactions between all of these processes are crucially altered by the metabolic milieu that characterizes diabetes mellitus, tipping the delicate balance towards atheroma formation, platelet aggregation and thrombus formation. This article will review these mechanisms and the effects of diabetes in the pathogenesis of vascular disease. PMID- 9212307 TI - Counterregulatory hormone and symptom responses to hypoglycaemia in diabetic children. AB - The hormonal responses to, and symptoms of, hypoglycaemia were investigated in 19 diabetic children (mean age 14.2 (SD 1.4) years, mean HbA1c 9.8 (SD 1.2)%) and 16 non-diabetic children (14.4(1.0) years) during a gradual reduction in plasma glucose with the glucose clamp technique. Plasma glucose was reduced from approximately 5.7 to approximately 2.6 mmol l(-1) in the diabetic children and from approximately 5.7 to approximately 2.9 mmol l(-1) in the non-diabetic children over 200 min. The mean glycaemic thresholds for adrenaline, and for autonomic and total symptom score, were similar in the diabetic and non-diabetic groups, and were found at plasma glucose levels between 3.4 and 3.7 mmol l(-1). The mean glucose levels which elicited increase of cortisol, growth hormone, and glucagon were lower (p < 0.01), and the mean incremental responses of adrenaline, cortisol, and glucagon were smaller in the diabetic than in the non-diabetic children. In the diabetic children, a correlation was found between Body Mass Index (BMI) and the hypoglycaemic thresholds for autonomic and total symptom scores (r = 0.64, p < 0.01 and r = 0.72, p = 0.001, respectively). We conclude that counterregulatory hormone responses are attenuated in diabetic as compared to non-diabetic children, whereas recognition of autonomic symptoms is similar in the two groups. Diabetic children with a higher BMI seem to have increased awareness of a declining plasma glucose level. PMID- 9212308 TI - Further evidence for a high incidence of nocturnal hypoglycaemia in IDDM: no effect of dose for dose transfer between human and porcine insulins. AB - We tested the hypothesis that transfer from porcine to human insulin causes a fall in nocturnal blood glucose and an increase in the frequency of hypoglycaemic episodes. Twenty IDDM patients (age 19-55, duration 3-36 years) used Velosulin and Insulatard twice daily for 12 weeks, double-blinded to species (human (H) or porcine (P)) in a randomized crossover study. Species was changed after 4 weeks' run-in and 4 weeks later, with insulin doses unchanged on transfer. Ten patients underwent each sequence (H/P/H or P/H/P) and were admitted on the first and eighth night after transfer for hourly blood glucose measurement (22.00-07.00). Biochemical hypoglycaemia (<3.5 mmol l(-1)) was observed on 39 of the 80 patient nights studied (48.75%). The number of episodes were similar during each night (H1 8, H8 10, P1 10, P8 11, p = 0.83). Total reported symptomatic episodes (H 51 vs P 73, p = 0.85), total HbA1 (H 9.8 +/- 0.3%, P 10.0 +/- 0.3%, p = 0.32) and daily insulin doses (H 0.63 +/- 0.04 units kg(-1) day(-1) vs P 0.63 +/- 0.05 units kg(-1) day(-1), p = 0.54) were not different. Despite an apparent fall in blood glucose levels from night 1 to 8 on transfer to human (AUC 82.3 +/- 7.8 vs 61.4 +/- 5.3 mmol.h l(-1), p < 0.05) but not porcine insulin (AUC 70.7 +/- 7.2 vs 70.1 +/- 7.5 mmol.h l(-1), p = 0.74), there was no difference when all 4 nights were considered together (p = 0.30). We conclude that dose for dose transfer to human insulin does not increase numbers of episodes of nocturnal or reported hypoglycaemia. PMID- 9212309 TI - Diabetic retinopathy assessed by fundus photography in Pima Indians with impaired glucose tolerance and NIDDM. AB - In a population-based epidemiological study, 991 Pima Indians with non-insulin dependent (Type 2) diabetes mellitus (NIDDM) and 288 without diabetes aged > or =15 years were examined for retinopathy by fundus photography with a 45 degrees fundus camera after mydriasis. The photographs were graded using a modified Airlie-House classification scheme. The associations of several factors with retinopathy were studied by logistic regression. Non-proliferative retinopathy was present in 11.2 % (19/169) subjects at the time of diagnosis of diabetes and in 8.3% (4/48) in newly diagnosed subjects who had a documented non-diabetic oral glucose tolerance test within 4 years prior to diagnosis of diabetes. The prevalence of retinopathy in subjects with impaired glucose tolerance was 12% (8/68). Retinopathy at the time of diagnosis of diabetes was significantly associated with lower body mass index and higher systolic blood pressure but not glycaemia. Retinopathy was present in 375 (37.8 %) diabetic subjects and 14 (5.2 %) non-diabetic subjects. Among all subjects with diabetes (duration 0-37 years), stepwise multivariate analysis showed non-proliferative retinopathy to be associated with duration of diabetes, mean blood pressure, fasting plasma glucose, treatment with insulin and albuminuria. Proliferative retinopathy was seen in 34 (2.7%) of diabetic and none of the non-diabetic subjects, and was associated with 2 h post-load glucose concentrations, as well as albuminuria, insulin treatment, younger age, and diastolic blood pressure. These data confirm the need for fundus examination at the time of diagnosis of diabetes and during long-term follow-up. Albuminuria and blood pressure are potentially modifiable risk factors and the impact of treating these on incidence and progression of diabetic retinopathy need to be assessed. PMID- 9212310 TI - Maternally inherited diabetes and deafness: prevalence in a hospital diabetic population. AB - Maternally inherited diabetes and deafness (MIDD) is a new sub-type of diabetes and results from an A to G substitution at position 3243 of the mitochondrial tRNA(leu(UUR)) gene. This mutation is also associated with a neurological syndrome (MELAS). Recent studies have screened carefully selected diabetic populations and have reported MIDD prevalence rates ranging from undetectable to 60%. The aim of this work was to determine the importance of this sub-type in clinical practice by screening a routine hospital diabetic population. A total of 1440 patients (IDDM and NIDDM) of North European extraction attending two hospital diabetes services were initially screened by questionnaire. This identified 445 patients with one or more features of MIDD and/or MELAS and these subjects were then genotyped. Two patients were identified with the mutation giving a prevalence rate of 0.13% for the whole study population, and 0.45% for the sample with phenotypic features of MIDD. In conclusion, therefore, the 3243 mutation is associated with the phenotypically distinct MIDD sub-type, but this is rare in the routine hospital diabetic population. PMID- 9212311 TI - Reproducibility and persistence of neural and adrenal autoantibodies in diabetic autonomic neuropathy. AB - The presence of autoantibodies to autonomic nervous tissue structures is a feature of patients with symptomatic diabetic autonomic neuropathy. It has not been established whether these autoantibodies cause, contribute to or simply reflect nervous tissue damage. Serum samples were tested for the presence of complement-fixing autoantibodies to adrenal medulla, vagus nerve, and sympathetic ganglion cells, to demonstrate: (a) reproducibility of the technique, (b) persistence of the antibodies, and (c) whether or not they occur in patients with non-insulin-dependent (Type 2) diabetes (NIDDM) with neuropathy. Examination of 37 samples, by different observers 2 years apart, revealed a high degree of concordance of both positive and negative results, demonstrating the method of testing to be highly reproducible. Re-testing of 37 patients (by analysing a second blood sample) between 0.5 and 2.7 years (mean 1.7 years) after their first test also demonstrated that antibodies, once present, normally persist; and that most patients initially negative remained so. Of 17 neuropathic NIDDM patients, 16 were negative for all three antibodies, indicating their rarity in this group of patients. PMID- 9212312 TI - Serum 7S domain of type IV collagen levels in essential hypertension and hypertensive type 2 diabetic patients. AB - Metabolic alteration of Type IV collagen occurs in micro- or macrovascular basement membrane of diabetic patients. Hypertension, a risk factor for clinical progression of diabetic vascular disease, may influence this metabolic alteration. The object of this study was to evaluate the serum 7S domain of type IV collagen (7S-collagen) levels in patients with essential hypertension and in Type 2 diabetic patients with or without hypertension and to investigate the relationship between the type IV collagen metabolism and the arterial blood pressure. Serum 7S-collagen levels in 18 patients with essential hypertension were significantly higher than in 24 normal subjects (4.2 +/- 0.5 vs 3.6 +/- 0.4 ng ml(-1) p < 0.01). Serum 7S-collagen levels in 28 normotensive diabetic patients (4.2 +/- 0.5 ng ml(-1)) were significantly higher than in normal subjects (p < 0.01). The serum 7S-collagen levels were significantly higher in 22 diabetic patients with hypertension (4.8 +/- 0.6 ng ml(-1)) than in the other groups. There was a significant correlation between the serum 7S-collagen levels and the systolic blood pressure in cases with essential hypertension (r = 0.59, p < 0.001) and in all diabetic patients (r = 0.52, p < 0.001), suggesting that elevation of the systolic blood pressure may influence the type IV collagen metabolism of vascular basement membrane. We conclude that the metabolic alteration of basement membrane occurring in patients with diabetes mellitus may worsen in the presence of high systolic blood pressure. PMID- 9212314 TI - Estimation of unascertained diabetes prevalence: different effects on calculation of complication rates and resource utilization. AB - The incidence and prevalence of insulin-dependent (Type 1) diabetes mellitus (IDDM) in populations are both well defined. In the more prevalent non-insulin dependent (Type 2) diabetes mellitus (NIDDM), which is responsible for the bulk of diabetes-related morbidity, true prevalence is uncertain because of delayed diagnosis and problems of definition, particularly with increasing age. Estimates therefore vary widely. We have previously presented evidence of increased relative probability of hospital admission for people with diabetes. These absolute and relative rates of admission were based on a large scale community derived prevalence for diabetes of 1.36%. Assuming that the true prevalence of diabetes is higher, recalculation of activity data in a sensitivity analysis suggests a theoretical maximum prevalence of diabetes of 5% in our population, since a higher value would imply less morbidity associated with diabetes than 'non-diabetes'. This approach identifies the possible range of unascertained diabetes in a population and defines it in functional terms as that state carrying any excess risk of admission for complications when compared to non diabetes. Higher estimates of prevalence have little impact on the calculation of overall resource use for diabetes, since the great majority of costs are related to fixed hospital activity for people with identified diabetes. The unascertained diabetes sub-group will cost little by comparison. Paradoxically, the tendency to use higher estimates of unascertained diabetes increases the denominator for calculation of complication rates and reduces both the absolute and relative risk of complications. This dilutes the epidemiological significance of diabetes in the aetiology of its related complications. PMID- 9212313 TI - Effect of insulin treatment on circulating islet amyloid polypeptide in patients with NIDDM. AB - The objective was to evaluate the effect of insulin treatment on circulating islet amyloid polypeptide (IAPP). Twelve patients with NIDDM and secondary failure were studied on oral agents and then switched to insulin treatment. Fasting and postprandial IAPP concentrations were measured on oral treatment and on insulin treatment. In 5 of the patients no postprandial concentrations were determined. In the 7 patients who were investigated both fasting and postprandially the fasting IAPP concentration was 6.5 +/- 1.2 pmol l(-1) (mean +/ SEM) during oral treatment with a rise to 13.5 +/- 3.1 90 min after breakfast (p = 0.028). On insulin treatment HbA1c decreased from 8.6 +/- 0.5 to 7.5 +/- 0.4% (p< 0.03) and plasma C-peptide concentration was significantly lowered (p< 0.01). There was a close correlation using simple regression between the per cent change of IAPP concentration and the per cent change of C-peptide concentration during this period (r = 0.88; p< 0.01). In the total patient material of 12 patients there was a significant correlation using simple regression analysis between per cent change of IAPP concentration and per cent change of C-peptide concentration using all 48 measurements available (r = 0.58: p< 0.001). These data suggest that secretion of IAPP is lowered when endogenous insulin secretion is lowered by administration of exogenous insulin in patients with NIDDM. Thus, if IAPP secretion has a pathogenetic role in the development of beta cell failure in NIDDM, insulin treatment might delay this deterioration. PMID- 9212315 TI - Hospital management of diabetic ketoacidosis: are clinical guidelines implemented effectively? AB - A study was undertaken in order to examine the quality of management of diabetic ketoacidosis (DKA) in a teaching hospital and to assess whether the introduction of clinical guidelines contributed to a satisfactory outcome. Data on presentation and management of 71 cases of DKA admitted in one calendar year (1994) were collected and analysed. Comparing the data to standards set in guidelines, inadequacies of clinical management were identified including delay in initiation of intravenous fluid replacement (greater than 30 min) and intravenous insulin (greater than 60 min) in 70% and 69% of cases, respectively; under-replacement with intravenous fluid in the first 24 h (less than 6.5 l) in 70% of cases, and insufficient intravenous potassium replacement (less than 70 mmol) in the first 24 h in 70% of cases. Suboptimal management of DKA may have contributed towards death in one of the three fatalities, and to morbidity in other patients. In 22.5% of cases (group 1) in whom the guidelines were alleged to have been followed, intravenous fluid, potassium, and insulin had been administered earlier and in larger quantities compared to the remaining cases (group 2). However, in most cases in group 1 the standards set by the guidelines were unfulfilled and the incidence of hypokalaemia, hypoglycaemia, and duration of in-patient stay did not differ from group 2. The treatment of DKA by non specialist general medical staff in a large teaching hospital was frequently inadequate and was associated with significant mortality and morbidity. The introduction of guidelines had moderately influenced the process of managing DKA but not the outcome, probably because of the low rate of their implementation by junior doctors. PMID- 9212316 TI - Conservative management of osteomyelitis in the feet of diabetic patients. AB - Experience of conservative management of osteomyelitis in a specialized, multidisciplinary, diabetic foot clinic was reviewed. The records of all patients attending the clinic over a 10-year period were examined retrospectively, and 22 patients with overt osteomyelitis were identified. Median age was 66 (31-87) years. In 12 cases the bone infection was a complication of a pre-existing ulcer; the most prevalent organism cultured from swabs was Staphylococcus aureus. The main site of infection was the first toe. The total duration of antibiotic treatment was 12 weeks (median, range 5-72), and clindamycin was the most commonly used oral agent. Four patients did not respond to initial conservative therapy and proceeded to amputation, while 1 patient responded clinically but had a recurrence of osteomyelitis at the same site 6 years later. In the remaining 17 patients resolution of osteomyelitis was achieved with conservative management over a median period of follow-up of 27 (range 5-73) months. The success of conservative therapy with prolonged courses of oral antibiotics challenges conventional advice that excision of infected bone is essential in the management of osteomyelitis affecting the foot in diabetes. PMID- 9212317 TI - Prevalence and incidence of NIDDM in Iceland: evidence for stable incidence among males and females 1967-1991--the Reykjavik Study. AB - This is the first large survey carried out in Iceland to estimate the prevalence and incidence of known and unknown non-insulin-dependent (Type 2) diabetes (NIDDM) among males and females, aged 34-79. The population in this survey was 9128 males and 9759 females born between 1907 and 1935 and examined in the prospective Reykjavik Study 1967-1991. Participants were invited from one to five times during the 24 years. The overall age-standardized prevalence (95% confidence limits) was 2.9% (2.5 to 3.3) for males and 2.1% (1.8 to 2.5) for females, aged 30-79, according to the European standard population. The overall annual age-standardized incidence rate per 100,000 was 377 (303 to 457) for males and 266 (212 to 320) for females, aged 35-74, standardized to the European population. Our study indicates that the prevalence of NIDDM is relatively low compared to other Nordic and western countries, and has not been increasing over the past 20 years. Furthermore, the incidence of NIDDM has not been changing during the past 20 years of follow-up among Icelandic males and females aged 34 79. PMID- 9212319 TI - Diabetes mellitus associated with mitochondrial myopathy and schizophrenia: a possible link between diabetes mellitus and schizophrenia. PMID- 9212318 TI - Nisoldipine blocks the increase of intracellular free calcium-ion concentration associated with elevated sodium-lithium countertransport activity in erythrocytes in patients with NIDDM. AB - To understand the mechanism by which elevated sodium-lithium countertransport activity (SLC) associates with increased intracellular free calcium-ion concentration ([Ca2+]i), we investigated the relationship between SLC and the effects of the extracellular Ca2+ concentration ([Ca2+]o) and a Ca2(+)-channel blocker, nisoldipine, on [Ca2+]i in erythrocytes from 48 patients with non insulin-dependent (Type 2) diabetes mellitus (NIDDM). There was a significant correlation between SLC and [Ca2+]i. Nisoldipine in the incubation medium significantly decreased [Ca2+]i, and there was a significant positive correlation between SLC and the degree of [Ca2+]i decrease. When the [Ca2+]o was elevated, [Ca2+]i was significantly increased, but nisoldipine almost completely suppressed this increase of [Ca2+]i. There was a significant positive correlation between SLC and the degree of the suppression. These data suggest that elevated SLC correlates with increased [Ca2+]i, and that the increased [Ca2i]i might be due to the increased Ca2+ influx through a dihydropyridine-sensitive Ca2+ pathway. PMID- 9212320 TI - Insulin therapy and risk of retinopathy in type 2 diabetes mellitus. PMID- 9212321 TI - The thrifty genotype hypothesis. PMID- 9212322 TI - Insulin therapy and its shortcomings - the need for new approaches. AB - After its discovery in 1921, insulin rapidly became established as a treatment for insulin-requiring diabetes mellitus (Type 1 and late-stage Type 2), providing effective symptom control and significant reductions in diabetes-associated mortality. However, within 30 years of insulin's discovery, physicians were faced with a new challenge - the treatment of the long-term complications of chronic hyperglycaemia. The Diabetes Control and Complications Trial provided clear evidence of the benefits of improved glycaemic control, but also highlighted the difficulties, such as an increased risk of hypoglycaemia, of attempting to achieve this using insulin as the only pharmacological agent. We now know that the pancreatic islet hormone, amylin, is also deficient in patients with Type 1 and late-stage Type 2 diabetes. It is possible that parallel replacement of both amylin and insulin may improve glycaemic control more smoothly in patients with diabetes, with less risk of hypoglycaemia, while still reducing the long-term sequelae of chronic hyperglycaemia. PMID- 9212324 TI - Role of amylin in nutrient intake - animal studies. AB - The pancreatic hormone amylin is co-secreted with insulin by beta-cells in response to nutrient intake. Studies performed in experimental animals have provided evidence that amylin may have several effects associated with carbohydrate metabolism. Amylin is a potent inhibitor of gastric emptying. This effect appears to require an intact vagus nerve and it is over-ridden by hypoglycaemia. These observations, coupled with the identification of putative amylin receptors in the area postrema of the hindbrain (a region implicated in the regulation of gastric motility) suggest that the effects of amylin on gastric emptying are mediated, at least in part, by the central nervous system. There is also evidence that amylin acts to inhibit food intake, an action which is distinct from its effects on gastric emptying. In addition, amylin has been shown to inhibit amino acid-stimulated glucagon secretion, suggesting that it may reduce endogenous glucose production in the postprandial period. PMID- 9212323 TI - Amylin: history and overview. AB - The presence of amyloid deposits in the pancreas was first described at the beginning of the 20th century. However, it was not until 1987 that the structure of the amylin molecule was identified. Amylin is a 37-amino-acid peptide hormone that is co-secreted with insulin by the pancreatic beta-cells in response to a nutrient stimulus. It is deficient in patients with Type 1 diabetes and elevated in patients in the early stages of Type 2 diabetes, a condition which is characterized by hyperinsulinaemia. Elevation of plasma amylin levels has also been described in patients with impaired glucose tolerance, obese subjects and in pregnant women with both normal glucose tolerance and gestational diabetes mellitus. However, it appears that deficiencies of amylin secretion appear before those of insulin in patients in the later stages of Type 2 diabetes. Early experimental studies suggested that amylin inhibits basal insulin secretion, and induces insulin resistance in skeletal muscle, leading to the hypothesis that it has a role in the aetiology of Type 2 diabetes. However, a number of more recent experimental studies have indicated that amylin is a third active pancreatic islet hormone that works with insulin and glucagon to maintain glucose homeostasis. Amylin appears to regulate glucose inflow to the circulation by influencing the rate of gastric emptying, and thus the rate at which meal-derived glucose enters the system, and also by inhibiting glucose release and hepatic glucose production in the postprandial period. PMID- 9212325 TI - Effects of amylin and the amylin agonist pramlintide on glucose metabolism. AB - Since the discovery of the pancreatic islet hormone amylin in 1987, its metabolic effects have been investigated in a number of studies in animals and humans. Data from some early animal studies suggested that amylin might be associated with the development of insulin resistance, but other studies found that amylin had no effect on insulin sensitivity. More recently, studies performed using the human amylin analogue pramlintide in patients with Type 1 diabetes found that the hormone has no influence on either insulin-stimulated glucose uptake or the restraining effect of insulin on hepatic glucose production during periods of euglycaemia. Furthermore, during insulin-induced hypoglycaemia, pramlintide appears to increase the plasma concentrations of cortisol and growth hormone, and to stimulate the release of the gluconeogenic substrate lactate by the skeletal muscles. Taken together with evidence that, in short-term studies, pramlintide improved glycaemic control in patients with Type 1 diabetes who were also treated with insulin, these data suggest that pramlintide may have a role in the management of patients with diabetes. However, longer-term studies are required to ascertain whether these findings are sustained over time. PMID- 9212326 TI - Amylin and the gastrointestinal tract. AB - There is increasing evidence that alterations in the rate of gastric emptying - both acceleration and slowing - are present in patients with diabetes mellitus. A number of different factors can influence the rate of gastric emptying. For example, a large meal volume, a high-fat meal, and the presence of high glucose concentrations will all slow the rate at which the contents of the stomach are emptied into the small intestine. Studies in spontaneously diabetic BB/Wistar rats have shown that administration of the pancreatic islet hormone amylin slows the rate of gastric emptying, and similar observations have been made in patients with Type 1 diabetes who received an intravenous infusion of the human amylin analogue pramlintide. The mechanism by which amylin slows gastric emptying is still unknown, but evidence from studies in animals suggests that it may influence the parasympathetic input to the stomach via neurons in the brainstem which regulate efferent activity in the vagus nerve. PMID- 9212327 TI - Amylin release during oral glucose tolerance test. AB - The role of amylin in the beta-cell dysfunction that occurs in patients with diabetes mellitus may be important. Amyloid deposits are found in the pancreata of subjects with Type 2 diabetes and may contribute to beta-cell death. It is therefore necessary to study amylin secretion and kinetics to determine whether elevated levels of the peptide are due to elevated secretion, reduced clearance or both. The aim of this study was to measure amylin dynamics during an oral glucose tolerance test (OGTT). We also used a mathematical model of beta-cell activity to assess the secretion and kinetics of C-peptide, insulin and amylin in humans during an OGTT. In particular, we were interested in characterizing the physiological meaning of one of the terms in the model, the amylin/C-peptide co secretion factor (sigma). The model has been used in several pathophysiological conditions and results indicate an elevated secretion and clearance of amylin in glucose-intolerant states. Amylin clearance has been found to be similar to that of C-peptide, and much slower than that of insulin. In this study, direct measurements of insulin and amylin secretion in five obese subjects yielded an estimate of the amylin/insulin co-secretion factor of 0.004 with a standard deviation (SD) of 0.002. The point estimate of hepatic clearance was 80 ml min( 1), which was much lower than that of insulin (507 +/- 94 ml min[-1]). In addition, the estimated hepatic clearance was not significantly different from zero given its high SD of 213 ml min(-1). The absence of hepatic extraction of amylin is therefore a plausible hypothesis, which is also supported by the similarity between amylin and C-peptide clearances. This observation characterizes the physiological meaning of sigma and suggests that this parameter is associated mainly with beta-cell secretion. PMID- 9212328 TI - Amylin and glycaemic regulation: a possible role for the human amylin analogue pramlintide. AB - Clinical studies with the human amylin analogue, pramlintide, suggest that it may help to improve glycaemic control in patients with diabetes mellitus using insulin. This has been demonstrated by reductions in postprandial glycaemic excursion, 24-h glucose profile and serum fructosamine concentrations following administration of pramlintide for periods of up to 28 days in patients with Type 1 diabetes. Additionally, preliminary studies with pramlintide in patients with Type 2 diabetes using insulin have indicated its ability to reduce postprandial hyperglycaemia in this population. Thus, this data set suggests a potential role for pramlintide as a partner to insulin for the optimization of glycaemic control in patients with diabetes using insulin. PMID- 9212329 TI - Partitioning and effects of silver in amended freshwater sediments. AB - Sediments that represented a wide range of characteristics were amended with silver compounds to observe partitioning and bioavailability. In laboratory studies, silver partitioning to particulates, sediment pore water, and overlying water was measured and bioavailability of silver was determined using Hyalella azteca in 10-day sediment toxicity tests. Three silver compounds were used as sources of silver for this study: silver nitrate, silver chloride, and silver thiosulfate complex. Sediment amendment procedures were adjusted as necessary depending on the characteristics of the individual compounds. Several sediment characteristics such as organic carbon, pH, redox, and acid volatile sulfides regulated silver partitioning and bioavailability. Bioavailability of silver was correlated with the overlying water concentration of silver. Ten-day LC50 values ranged from 1.62 to 379.7 mg Ag/kg for H. azteca exposed to sediments amended with AgNO3. In laboratory experiments, silver chloride and silver thiosulfate were orders of magnitude less toxic and bioavailable than silver nitrate, with 10 day LC50 values greater than the highest concentrations of AgCl and silver thiosulfate complex amended to sediments (2560 and 1125 mg Ag/kg, respectively. PMID- 9212330 TI - Sensitivity of mouse lymphoid and nonlymphoid organs to Silesian air pollutants. AB - The toxic effect of Silesian air pollutants to mouse organs was examined. Histological changes were found in the examined lymphoid organs (thymus, spleen) as well nonlymphoid organs (liver, kidneys). The alterations in weight indexes of lymphoid organs were also observed. Considerable changes in cellularity, weight index, and histology of the thymus in the mice exposed to air pollutants suggest the atrophy of this organ, which may lead to extrathymic T-cell differentiation and even acceleration of thymocytes maturation, which may lead to certain allergic or auto-immune pollutants of all investigated mouse organs in the following order: thymus, liver, kidneys, and spleen. PMID- 9212331 TI - Similarities and differences between the massive eel (Anguilla anguilla L.) devastations that occurred in Lake Balaton in 1991 and 1995. AB - In the past few years, two massive eel (Anguilla anguilla L.) devastations occurred in Lake Balaton, Hungary. In 1991, 300 tons of eel perished in the western basin of the lake, while in the summer of 1995 30 tons of eel died in the eastern part of the lake. Investigations carried out to find the causes of these ecocatastrophes included measurements of certain biochemical parameters: the blood sugar level, and the acetylcholinesterase (AChE, EC 3.1.1.7), lactate dehydrogenase (LDH, EC 1.1.2.3), glutamic-oxaloacetic transaminase (GOT, EC 2.6.1.1) and glutamic-pyruvic transaminase (GPT, EC 2.6.1.2) activities in the blood serum of the collected eels. In both 1991 and 1995, deltamethrin (DM), the active ingredient of the insecticide K-OTHRIN 1 ULV used against mosquitoes, was detected in the eels; in 1995 it was demonstrated in several other animal species, i.e., bream (Abramis brama L.), pike perch (Stizostedion lucioperca L.), and the common gull (Larus canus), and in sediment samples from the lake. Additionally, laboratory experiments were carried out to study the effects of DM on eels. In 1991, eels were collected from the western (the site of the devastation) and eastern basins of the lake. The eels from the eastern basin were used as controls. At that time, the AChE activity in the blood serum of the eels from the western basin was significantly inhibited compared to that in animals from the eastern basin (P < 0.05, Student t test). Eels from the western part of the lake had GOT and GPT levels 20 and 100%, respectively, higher than those of eels from the eastern part of the lake. The blood glucose level was much higher in the eels from the affected area of the lake as compared to those from the eastern part. The brain and liver of the eels contained DM residues at 20 micrograms/kg wet tissue (Gonczy, 1992). Gonczy suspected that one of the causes of the massive eel loss in 1991 was the presence of DM in the fish. In 1995, when the eel devastation occurred in the eastern basin, moribund and surviving eels were collected from this part of the lake. The AChE activity was significantly inhibited in the blood serum of the dying eels as compared to that in surviving animals (P < 0.05, Student t test). The blood glucose content exhibited a difference too: it was 2.5 times higher in the dying eels than in the surviving ones. A huge increase in the LDH level was measured in the dying eels, indicating damage to different muscle tissues to an extent never observed previously. The GOT activities of the serum were twice as high in the dying eels as in the living fish. The GPT was not significantly changed in the serum of dying eels as compared to the surviving animals. DM was detected in different tissue samples of the dying eels: 2.7-18.5 micrograms/kg in the liver, 9.0-31.1 micrograms/kg in the gill, and 3.0 micrograms/kg wet tissue in the muscle. DM residues were found in tissue samples from other animals, in the following concentrations: 0.4 micrograms/kg in bream, 2.1 micrograms/kg in pike perch, 1.1 micrograms/kg wet tissue in dead gulls. The sediment samples collected from different places and at different times contained DM in a concentration of 5.5-30.0 micrograms/kg wet sediment at the time of the eel deaths, while the sediment samples collected from the same places a month later still contained DM at 7.0-8.8 micrograms/kg wet sediment. Laboratory experiments with the insecticide K-OTHRIN 1 ULV revealed that 1.0 microgram/liter of its active ingredient, DM, caused the death of 50% of the eels after an incubation time of 96 hr. In the liver of the dead eels, DM was detected at 2.9-20.0 micrograms/kg wet tissue. All the above-mentioned changes and the DM residue detected in the eels appear to demonstrate the contribution of DM in the severe eel devastation. This finding on the ecological risk of such types of insecticides might be useful in their further application. PMID- 9212332 TI - Uptake and transformation of benzene and toluene by plant leaves. AB - The [1-6(14)C]benzene and [1-(14)C]toluene vapors penetrate into hypostomatous leaves of Acer campestre, Malus domestica, and Vitis vinifera from both sides, whereas hydrocarbons are more intensively absorbed by the stomatiferous side and more actively taken up by young leaves. Benzene and toluene conversion in leaves occurs with the aromatic ring cleavage and their carbon atoms are mainly incorporated into nonvolatile organic acids, while their incorporation into amino acids is less intensive. Intact spinach chloroplasts oxidize benzene, and this process is strongly stimulated in light. Oxidation of benzene by spinach chloroplasts or by enzyme preparation from spinach leaves is almost completely inhibited by 8-oxyquinoline or sodium diethyldithiocarbamate, and slightly affected by alpha, alpha'-dipyridyl. Benzene oxidation by enzyme preparation is significantly stimulated by NADH and NADPH; in their presence, the benzene hydroxylation product, phenol, is formed in a determinable amount. It is supposed that the enzyme performing the first step of oxidative transformation of benzene in plant leaves contains copper as the prosthetic group. PMID- 9212334 TI - Toxicity of increased amounts of chemicals and the dose-response curves for heterogeneous microbial populations in soil. AB - The paper deals with the interpretation and classification of dose-response curves in order to understand the way in which the heterogeneous soil microbial population behaves under chemical stress. The evaluation is based on a set of about 500 toxicity tests, in which geometrically increasing doses of toxicants were applied to soil samples. The responses of the microflora were measured by various methods, e.g., Fe(III) reduction, substrate induced respiration, arginine ammonification, and several enzyme activities. The data reveal that microbial populations in soil react more complexly than homogeneous groups of test subjects which are common in classical toxicology. The diverse types of dose-response curves are attributed to a varying sensitivity of different parts of the soil microflora and influences of the habitat soil. A proposal for the interpretation and classification of microbial dose-response curves is presented. Four basic types of dose-dependent effects and several combined sequences of them can describe the reaction patterns found up to now. Since experiments with heterogeneous populations are lacking in classical toxicology, the results can be used as a key for further research regarding the toxicity of chemicals against plant, animal, and human populations. PMID- 9212333 TI - Toxicity of acid-sulfate soil leachate and aluminum to embryos of the Sydney Rock oyster. AB - The toxicity of leachate water from acid-sulfate soil to the early embryonic development of the Sydney Rock oyster, Saccostrea commercialis, was assessed. Concentrations of acid-sulfate soil leachate water as low as 3.3% in seawater were found to decrease the normal development of oyster embryos after 48 hr exposure, and this effect could not be attributed to any significant change in pH or salinity. An EC50 value for the acid-sulfate soil leachate water of 2.5 to 2.9% in seawater was obtained, and the no observed effect concentration was determined at a concentration of 2% in seawater. In tests conducted with aluminum added to seawater, a significant decrease in the percentage of embryos developed to the D-veliger stage occurred at concentrations of 150 micrograms/liter and greater, with no effects at 100 micrograms/liter. An EC50 of 225 micrograms/liter for the effect of added aluminum on embryo survival was obtained and all embryos showed developmental abnormalities at concentrations of 400 micrograms/liter and greater. A significant decrease in the embryonic development occurred when the fertilized eggs were incubated in pH-adjusted seawater at pH values < or = 6.75, but no significant effects were found at pH 7.0 or above. Since aluminum was present in high concentrations in the acid-sulfate soil leachate water, it was concluded that aluminum was the main toxicant in the acid-sulfate water that disrupted the oyster embryonic development. PMID- 9212335 TI - A comparison of acute mortality and population growth rate as endpoints of toxicological effect. AB - The objective of this study was to determine how closely acute (72-hr) lethal concentration estimates developed from probit analysis compared to the demographic toxicological endpoints, net reproductive rate (Ro), the intrinsic rate of increase (rm), and realized fecundity (Ux), in terms of predicting effects of pesticides on populations. Lethal and sublethal effects of the insecticide imidacloprid on the arthropod Acyrthosiphon pisum Harris (pea aphid) were determined for populations exposed to foliar-sprayed broad bean Vicia faba L. (variety Banner). An examination of Ro indicated that sublethal effects were occurring that reduced reproduction. However, by looking at the mean number of offspring produced per surviving female and Ux, it was determined that the reduction in Ro was entirely due to acute mortality and a reduction in life span. Also, exposure to increasing concentrations of imidacloprid did not cause a shift in either the day of initial reproduction or the day of peak reproduction. Therefore, this pesticide caused no sublethal effects on reproduction and, as such, a lethal concentration estimate should have been a good predictor of effect at the population level. However, the 72-hr lethal concentration estimate was not a good predictor of effect of this pesticide on population growth. Populations exposed to the 72-hr LC60 were able to maintain rates of population increase (rm = 0.224) similar to those of the control (rm = 0.295). The data indicate that the reason for the discrepancy between acute lethal concentration estimates and population growth was that surviving individuals were able to sustain heightened rates of reproduction following acute exposure to imidacloprid. The ability of surviving individuals to maintain these high reproductive rates allowed them to compensate for losses and act as reservoirs for future reproduction. It is not possible, using acute mortality estimates alone, to predict this "reservoir effect," and therefore not possible to predict how a population's growth rate will respond or change based on this endpoint. Thus this would suggest that the assessment of a xenobiotic based solely on acute mortality estimates will lead to flawed conclusions about a population's exposure response. PMID- 9212336 TI - Application of multiple bioindicators to differentiate spatial and temporal variability from the effects of contaminant exposure on fish. AB - Bioindicators of fish health were measured from 1989 through 1994 in populations of redbreast sunfish (Lepomis auritus) at three sites within a stream receiving inputs of mixed contaminants from an industrial facility, and at two reference sites. Bioindicator responses differed for fish from contaminated and reference sites throughout the study period, but temporal trends at contaminated sites reflected improved water quality associated with pollution-control efforts. Temporal variability unrelated to contaminant exposure strongly influenced the response of bioindicators at both reference and contaminated sites, but spatial variability rarely influenced these responses. Temporal variability was less influential on slower responding indicators at higher levels of biological organization. The diverse response characteristics of indicators increased the ability to differentiate natural from anthropogenic sources of variability. Integrated bioindicator responses were compared among site-year groups by multivariate canonical analysis. The primary canonical variable, associated with differences in growth, exhibited trends consistent with the timing of pollution control efforts. Two indicators of contaminant exposure (7-ethoxyresorufin O deethylase activity and polychlorinated biphenyls in fish muscle) also exhibited trends consistent with the timing of pollution-control efforts, but the likelihood of possible mechanistic linkages cannot be assessed with available data. PMID- 9212337 TI - Effects of cadmium and mercury on ovarian maturation in the red swamp crayfish, Procambarus clarkii. AB - In vivo mercury significantly inhibited ovarian maturation in the red swamp crayfish, Procambarus clarkii. 5-Hydroxytryptamine (5-HT) induced ovarian maturation in vivo. Cadmium and mercury inhibited this 5-HT-induced maturation. Ovarian explants incubated with mercury and either brain or muscle demonstrated significant inhibition of [14C]leucine incorporation into ovarian proteins compared to the corresponding groups incubated without mercury. In the absence of mercury the brain, which contains a gonad-stimulating hormone (GSH), induced significantly more incorporation of this amino acid than occurred in the ovaries incubated with muscle. These metals may have exerted their inhibitory effects by directly inhibiting protein synthesis in the ovaries, inhibiting 5-HT-stimulated GSH release, and preventing the ovaries from responding to this hormone. PMID- 9212338 TI - Internal lethal concentrations of halobenzenes with fish (Gambusia affinis). AB - The internal lethal concentrations is a potential measure of toxicity which could be usefully applied in environmental toxicology and risk assessment. Using halobenzenes, which are common environmental contaminants, and represented test compounds, experiments were conducted in aquaria with the mosquitofish (Gambusia affinis). The average internal lethal concentration (ILC50) for four representative halohydrocarbons, 1,4-diBB, 1,2,3-triCB, 1,2,4-triBB, and pentaCB, were consistent with those previously observed, i.e., 2.3-8.3 mmol kg-1 fish over exposure time periods of 10.4 to 633 hr. However, the ILC50 for all the compounds is not constant but decreases with increasing exposure time period with a mean first-order rate constant of (4.21 +/- 0.70) x 10-3 hr-1. The time dependency of the ILC50 is inconsistent with the critical internal concentration hypothesis which requires the ILC50 to reach a constant critical value when lethality occurs. The life expectancy of the fish from the beginning of chemical exposure could possibly be related to the ILC50-exposure time relationship. PMID- 9212340 TI - Environmental hazard of selenium in the Animals La Plata water development project. AB - A hazard assessment of selenium was conducted for the Animas La Plata Project, a multiple-use water development proposed for Colorado and New Mexico by the United States Bureau of Reclamation. A published protocol for aquatic hazard assessment of selenium was applied to environmental monitoring data to assess current threats to biota in the water supply rivers (Animas, La Plata, and Mancos Rivers). Hazard evaluation were also made for two proposed reservoirs (Ridges Basin and Southern Ute Reservoirs) based on estimated concentrations of selenium. The assessment protocol indicated moderate hazard in the Animas and La Plata Rivers, and high hazard in the Mancos River and both of the proposed reservoirs. These ratings indicate that the risk of selenium poisoning in fish and aquatic birds is substantial. Moreover, the geology and climate of this site make it prone to irrigation-induced selenium contamination of water and biota. The water supplies already contain dangerously high concentrations of selenium that may increase further due to agricultural irrigation drainage. The stage is set for significant environmental problems unless a development scenario can be devised that will effectively reduce ecological risks. PMID- 9212339 TI - Environmental degradation of polyacrylamides. II. Effects of environmental (outdoor) exposure. AB - The environmental fate of a polyacrylamide thickening agent (PATA), formulated without and with a glyphosate-surfactant herbicide (GH), was examined under various environmental situations: formulation in surface water and ground water, volatility, and soil mobility. Environmental Fate of PATA in Surface Water and Ground Water: PATA was formulated at four concentrations in distilled-deionized water, three surface water samples, and two ground water samples, without and with a GH. Solutions were placed in glass bottles, covered with plastic wrap, and exposed to environmental (outdoor) conditions for 6 weeks. Acrylamide and ammonium concentration, pH, and bacterial and fungal populations were measured weekly. All solutions in this portion of the study had a homogeneous milky appearance but the conclusions of the study were nearly transparent. The results of this study suggest that polyacrylamide can degrade to acrylamide under environmental conditions. Statistically, there was no linear correlation between the various parameters measured. Volatility: PATA was formulated without and with GH. Each solution plus an acrylamide standard (positive control) was placed in a glass beaker and exposed to environmental (outdoor) conditions for 6 days. Acrylamide concentration, ammonium concentration, pH, and solution volume were measured daily. Acrylamide and ammonium concentrations increased during the study in all formulations, except when solutions evaporated to dryness. pH did not change greatly over the course of the study for these samples. Those solutions containing PATA had a homogeneous milky appearance but by the conclusions of the study were nearly transparent. This suggests a physical structural change in the polymer. Soil Mobility: PATA formulated with GH was also applied to soil columns and soil boxes containing sand, Eudora sandy loam, Eudora sandy clay, and Kohola silt loam. Acrylamide could be detected by Day 2 in all soil columns. Acrylamide could not be detected in the runoff of any of the soil boxes. PMID- 9212341 TI - Additivity in microbial toxicity of nonuniform mixtures of organic chemicals. AB - Microbial toxicity of nonuniform mixtures of selected synthetic organic chemicals in several proportion is evaluated. Toxicity is quantified by the inhibition of oxygen uptake rate of a surrogate microbial text culture as measured by a respirometer. The joint toxic effects of the chemicals are analyzed for simple addition using toxic units (TU) and similarity parameters (lambda). A new approach is proposed to assign acceptance limits to sigma(TU)i and lambda to account for experiment errors and variances. Based on this approach, the joint toxic effects of 16 chemicals evaluated in this study in 14 different mixtures were found to be simply additive. Predictions of component concentration based on simple additivity agreed with the measured values within an average factor of error of 1.4. PMID- 9212342 TI - Pharmacokinetic analysis of free radicals by in vivo BCM (Blood Circulation Monitoring)-ESR method. AB - In pharmacokinetic studies, a variety of analytical method including radioisotopic detection and HPLC (high performance liquid chromatography) has been used. In the present investigation, we developed in vivo BCM (Blood Circulation Monitoring)-ESR method, which is a new technique with a conventional X-band ESR spectrometer for observing stable free radicals in the circulating blood of living rats under anaesthesia. Both 5-(PROXYL derivatives) and 6-(TEMPO d derivatives) membered nitroxide spin probes with various types of substituent functional group were used. After physico-chemical properties of the spin probes such as hyperfine coupling constant (A-value), g-value and partition coefficient as well as chemical stability of the compounds in the fresh blood were obtained, the in vivo BCM-ESR method was performed in normal rats. Several pharmacokinetic parameters such as half-life of the probes, distribution volume, total body clearance and mean residence time were obtained and discussed in terms of their chemical structures. In addition, clearance of a spin probe was related to the urine concentration. The BCM-ESR method was found to be very useful to observe free radicals at the real time. By time-dependent ESR signal decay of spin probes, pharmacokinetic parameters were obtained. PMID- 9212343 TI - Superoxide radical generation in peroxisomal membranes: evidence for the participation of the 18 kDa integral membrane polypeptide. AB - Peroxisomes were isolated from pea (Pisum sativum L.) leaves and the peroxisomal membranes were purified by treatment with Na2CO3. The production of superoxide radicals (O2) induced by NADH was investigated in peroxisomal membranes from intact organelles incubated with proteases (pronase E and proteinase K). Under isoosmotic conditions, in the presence of pronase E, the production of O2-. radicals was inhibited by 80%. SDS-PAGE of peroxisomal membranes after protease treatment demonstrated a decrease in the 18-kDa PMP. This suggests that this polypeptide has a small fragment exposed to the cytosolic side of the peroxisomal membrane which is essential for O2-. production. The 18-kDa PMP was purified by preparative SDS-PAGE and in the reconstituted protein the NADH-driven production of O2-. radicals was investigated. The isolated polypeptide showed a high generation rate of superoxide (about 300 nmol O2-. x mg-1 protein x min-1) which was completely inhibited by 50 mM pyridine. The 18-kDa PMP was recognized by a polyclonal antibody against Cyt b5 from human erythrocytes. The presence of b type cytochrome in peroxisomal membranes was demonstrated by difference spectroscopy. Results obtained show that in the NADH-dependent O2-. radical generating system of peroxisomal membranes, the 18-kDa integral membrane polypeptide, which appears to be Cyt b5, is clearly involved in superoxide radical production. PMID- 9212344 TI - Effect of repeated exercise on urinary 8-hydroxy-deoxyguanosine excretion in humans. AB - The purpose of this study was to investigate the effect of repeated exercise on oxidative damage to DNA in 10 well trained long distance runners who participated in an 8-day training camp. The average running distance during the camp was 30 +/ 3 km/day. The amount of urinary 8-hydroxy-deoxyguanosine (8-OHdG) excretion was used to estimate the oxidative DNA damage. Urine samples were collected for both a 3-day control period as well as throughout the camp. Blood samples were drawn after overnight fasting both before and after the camp. Urinary 8-OHdG excretion was significantly increased during the camp compared to the control period (265.7 +/- 75.5 vs. 335.6 +/- 107.4 pmol/kg/day, P < 0.05). The content of 8-OHdG in the lymphocyte DNA on the day after finishing the camp did not differ from that before the camp. Plasma TBARS, LDH, CK, CK-MB, and myoglobin significantly rose after the camp (P < 0.05). The plasma beta-carotene levels tended to rise after the camp, while the plasma alpha-tocopherol levels increased significantly after the camp (P < 0.05). These results indicate that repeated exercise augments oxidative stress and the DNA is also injured by exercise-induced reactive oxygen species. However, the oxidative damage to DNA is not accumulated by consecutive exercise, although it is sustained as long as the exercise is repeated. PMID- 9212345 TI - Pitfalls in a method for assessment of total antioxidant capacity. AB - A relatively simple and widely applied method for quantitating the total antioxidant capacity of body fluids and drug solutions based on the absorbance of the ABTS radical cation was evaluated. In this assay, the end-point is an antioxidant-induced decrease in absorbance at a fixed time. This decrease is used as an index of total antioxidant capacity. It is shown that Trolox, potassium cyanide and quercetin all decrease the absorbance of ABTS radical cations at a fixed time, but by different mechanisms. Trolox scavenges the ABTS radical, potassium cyanide inhibits radical formation, while quercetin acts by both mechanisms. Using this method antioxidant capacity may be overestimated, due to both a scavenger effect and an effect on the rate of ABTS oxidation. To distinguish between these effects, a post-addition assay was used in which the sample is added when the formation of radicals is stable. Using post- addition assay conditions enables discrimination between effects on radical scavenging and on the radical formation, two major mechanisms for antioxidant action. In extrapolating the results to an in vivo situation it should be questioned: (i) whether the peroxidase process does indeed mimic the process of radical formation in vivo, and (ii) whether the ABTS radicals do resemble the radical species involved in an in vivo situation. Results obtained in the ABTS radical-based methods should therefore be reviewed critically before the antioxidant capacity can be assessed. PMID- 9212346 TI - Effect of endurance exercise on the tissue 8-hydroxy-deoxyguanosine content in dogs. AB - The purpose of this study was to investigate the effect of endurance exercise on both the tissue and lymphocyte 8-hydroxy-deoxyguanosine (8-OHdG) content. Six dogs ran on a treadmill for 7 hours. Another six dogs were assigned to a sedentary control group. The exercised dogs were sacrificed immediately after exercise and the counterpart of the sedentary group was also sacrificed at the same time. The brain, lung, liver, spleen, kidney, jejunum, colon, diaphragm, heart, splenius muscle, and the medial and lateral portion of gastrocnemius muscle samples were then collected. Lymphocytes were sampled before and after exercise in the exercised dogs. The 8-OHdG content of lymphocyte DNA was found to significantly decrease after exercise (0.57 +/- 0.19 vs 0.33 +/- 0.10/deoxyguanosine (dG) x 10(5), P < 0.05). The colon was the only tissue which showed a significant decrease in the content (0.83 +/- 0.24 vs 0.54 +/- 0.15/dG x 10(5), P < 0.05). No tissue except for the colon showed any significant changes after exercise. These results therefore indicate that, immediately after endurance exercise, an augmented repair mechanism might thus play a role in the decrease of 8-OHdG in the lymphocytes and the colon, while the 8-OHdG generation might be counterbalanced by its repair in other tissues. PMID- 9212347 TI - A new spiro[indoline-naphthoxazinic] spin trapping agent. AB - The synthesis and characterisation of a new spin trapping agent of the spiro[indoline-naphthoxazinic] family is reported. The EPR results of spin trapping experiments are also described, which indicate the ability of this nitrone to react with carbon-, oxygen- and sulphur-centered radicals. PMID- 9212348 TI - Alternative mechanisms for hydroperoxide-induced DNA single strand breakage. AB - The results presented in this study point out the existence of similarities as well as differences in the DNA-damaging effects of organic vs. inorganic hydroper oxides in human myeloid leukemia U937 cells. On the one hand, the formation of DNA single strand breaks (SSBs) induced by either hydrogen peroxide (H2O2) or tert-butylhydroperoxide (tBu-OOH) was prevented by iron chelators, was not affected by antioxidants or glucose omission before and during peroxide exposure and was enhanced by prior catalase depletion. Furthermore, H2O2- and tBu-OOH induced DNA strand scission were also detected after treatment at 0 degree C. On the other hand, H2O2, but not tBu-OOH or cumene hydroperoxide (cum-OOH), produced DNA strand scission in isolated nuclei and post-lysed DNA samples. In addition, lowering the basal intracellular calcium concentration with ethylene glycol bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA) markedly reduced the DNA-damaging efficiency of tBu-OOH while promoting only a slight decline in the number of DNA SSBs induced by H2O2. Taken together, these results are consistent with the commonly held theory that DNA damage caused by H2O2 is mediated by the formation of hydroxyl radicals. tBu-OOH-induced DNA single strand breakage appears to involve both the formation of H2O2 and a rise in cytosolic calcium ions. PMID- 9212349 TI - Free radical transients in photobleaching of xanthophylls and carotenes. AB - Carotenoids in chloroform and carbon tetrachloride photobleach upon nanosecond laser flash photolysis in two steps: instantaneously and in a second-order reaction. The rate constant for second-order reaction (first-order in a solvent derived radical and first-order in (excess) carotenoid) is largest for carotenes (9.8.10(8) M-1 for beta-carotene), intermediate for hydroxylated carotenoids, and smallest for carbonyl containing carotenoids (1.0.10(8) M-1 S-1 for astaxanthin) in chloroform at 20 degrees C. Near infrared absorbing transients are formed concomitant with photobleaching in chloroform (not detected in carbon tetrachloride). A species formed instantaneously is tentatively identified as either a carotenoid/solvent adduct or an ion-pair. A second species is formed by decay of instantaneously formed species and is identified as the carotenoid radical action. This species is formed in a first-order reaction with a rate constant of approx. 5.10(4) S-1 and absorbing at longer wavelength than the precursor. The lifetime (second-order decay) of the intermediates appears to be longest for the carotenoids with the longest conjugated system. The results indicate that carotenes are better antioxidants than xanthophylls as the carotenes, at least in the present lipophilic solvents, react faster with free radicals. PMID- 9212350 TI - Will the 'good fairies' please prove to us that vitamin E lessens human degenerative disease? AB - Recent research about the role of free radical derivatives of oxygen and nitrogen in biological systems has highlighted the possibility that antioxidants, such as vitamin E, that prevent these processes in vitro may be capable of carrying out a similar function in living organisms in vivo. There is increasing evidence that free radical reactions are involved in the early stages, or sometimes later on, in the development of human diseases, and it is therefore of particular interest to inquire whether vitamin E and other antioxidants, which are found in the human diets, may be capable of lowering the incidence of these diseases. Put simply, the proposition is that by improving human diets by increasing the quantity in them of antioxidants, it might be possible to reduce the incidence of a number of degenerative diseases. Of particular significance to these considerations is the likely role of the primary fat-soluble dietary antioxidant vitamin E in the prevention of degenerative diseases such as arteriosclerosis, which is frequently the cause of consequent heart attacks or stroke, and prevention of certain forms of cancer, as well as several other diseases. Substantial evidence for this proposition now exists, and this review is an attempt to give a brief account of the present position. Two kinds of evidence exist; on the one hand there is very substantial basic science evidence which indicates an involvement of free radical events, and a preventive role for vitamin E, in the development of human disease processes. On the other hand, there is also a large body of human epidemiological evidence which suggests that incidence of these diseases is lowered in populations having a high level of antioxidants, such as vitamin E, in their diet, or who have taken steps to enhance their level of intake of the vitamin by taking dietary supplements. There is also some evidence which suggests that intervention with dietary supplements of vitamin E can result in a lowered risk of disease, in particular of cardiovascular disease, which is a major killer disease among the developed nations of the world. The intense interest in this subject recently has as its objective the possibility that, by making some simple alterations to dietary lifestyle, or by enhancing the intake of vitamin E by fortification of foods, or by dietary supplements, it may be possible to reduce substantially the risk of a large amount of common, highly disabling human disease. By this simple means, therefore it may be possible to improve substantially the quality of human life, in particular for people of advancing years. PMID- 9212351 TI - Ascorbic acid inhibits lipid peroxidation but enhances DNA damage in rat liver nuclei incubated with iron ions. AB - In this report we studied DNA damage and lipid peroxidation in rat liver nuclei incubated with iron ions for up to 2 hrs in order to examine whether nuclear DNA damage was dependent on membrane lipid peroxidation. Lipid peroxidation was measured as thiobarbituric acid-reactive substances (TBARS) and DNA damage was measured as 8-OH-deoxyguanosine (8-OH-dG). We showed that Fe(II) induced nuclear lipid peroxidation dose-dependently but only the highest concentration (1.0 mM) used induced appreciable 8-OH-dG. Fe(III) up to 1 mM induced minimal lipid peroxidation and negligible amounts of 8-OH-dG. Ascorbic acid enhanced Fe(II) induced lipid peroxidation at a ratio to Fe(II) of 1:1 but strongly inhibited peroxidation at ratios of 2.5:1 and 5:1. By contrast, ascorbate markedly enhanced DNA damage at all ratios tested and in a concentration-dependent manner. The nuclear DNA damage induced by 1 mM FeSO4/5 mM ascorbic acid was largely inhibited by iron chelators and by dimethylsulphoxide and mannitol, indicating the involvement of OH. Hydrogen peroxide and superoxide anions were also involved, as DNA damage was partially inhibited by catalase and, to a lesser extent, by superoxide dismutase. The chain-breaking antioxidants butylated hydroxytoluene and diphenylamine (an alkoxyl radical scavenger) did not inhibit DNA damage. Hence, this study demonstrated that ascorbic acid enhanced Fe(II)-induced DNA base modification which was not dependent on lipid peroxidation in rat liver nuclei. PMID- 9212352 TI - Acquired isolated factor VII deficiency during sepsis. AB - Isolated acquired factor VII deficiency is uncommon. We report 11 cases of acquired factor VII deficiency associated with severe systemic sepsis. All patients initially displayed a heterozygous-like factor VII deficiency confirmed by both clotting and amidolytic assays, associated with low factor VII antigen levels, and increased haemostasis markers (D-dimers, prothrombin fragments 1.2, thrombin-antithrombin complexes). After sepsis recovery, normal factor VII levels were evidenced. Isolated factor VII consumption or proteolytic degradation by leucocyte proteases can be evoked, but the mechanism of acquired factor VII deficiency during sepsis remains to be elucidated. The knowledge of this syndrome should avoid false diagnosis of congenital factor VII deficiency. PMID- 9212353 TI - Urinary procoagulant activity and tissue factor levels in patients with diabetes mellitus. AB - We characterized a factor important for the normal procoagulant activity in human urine and measured urinary tissue factor (TF) and thrombomodulin (TM) levels in patients with diabetes mellitus (DM). Urine shortened normal plasma-based clotting time, which is inhibited by antibody to the human TF. TF in urine presents in membrane-nonassociated and associated forms. The urine TF antigen levels in DM patients were significantly lower than those in healthy subjects, particularly in DM patients without retinopathy. Levels of TM in patients with DM and normal subjects were similar. Although urinary TF might be necessary to repair tissue injury of damaged sites in the excretory pathway of urine, the clinical relevance of the reduced TF in the urine needs further investigation. PMID- 9212354 TI - Low molecular weight heparin and unfractionated heparin for prevention of thrombo embolism in general surgery: a meta-analysis of randomised clinical trials. AB - Low molecular weight heparin (LMWH), unfractionated heparin (UFH) and warfarin were compared with respect to efficacy and safety in the prevention of thrombo embolism in general surgery. Meta-analysis (MA) with a priori definition of the MA protocol was used to combine the results from randomised trials with patients who underwent general surgery and deep-vein thrombosis (DVT) prophylaxis with LMWH, UFH or warfarin. Forty-four studies were identified for assessment and 33 were included, however, none for warfarin. For efficacy (DVT and pulmonary embolism) and major bleeding, no significant difference between the LMWH- and UFH treated groups was demonstrated. The relative risk of minor bleedings for LMWH versus UFH was 0.75 (0.64-0.88; 95% confidence interval) and is significant (p < 0.05). Within the limitations of the MA, LMWH and UFH did not differ significantly in terms of prevention of thrombo-embolism, but LMWH had a significantly better safety profile. On this basis, LMWH may be preferable to UFH in the prevention of thrombo-embolism in general surgery. PMID- 9212355 TI - Efficacy and safety of low molecular weight heparin, unfractionated heparin and warfarin for thrombo-embolism prophylaxis in orthopaedic surgery: a meta-analysis of randomised clinical trials. AB - The efficacy and safety of low molecular weight heparin (LMWH), unfractionated heparin (UFH) and warfarin for prophylaxis of thrombo-embolism in orthopaedic surgery were compared using meta-analysis techniques. Twenty-two studies were included, 2 of which compared LMWH to warfarin. The mean probabilities to develop deep-vein thrombosis (DVT), pulmonary embolism and major and minor bleeding using UFH were: 0.21 (95% confidence interval, CI: 0.18-0.24); 0.01 (95% CI: 0.01 0.02); 0.05 (95% CI: 0.03-0.07), and 0.19 (95% CI: 0.17-0.22), respectively. The relative risk (RR) of DVT for LMWH vs. UFH was 0.76 (95% CI: 0.60-0.91), p < 0.05 and for LMWH vs. warfarin 0.78 (95% CI: 0.69-0.87), p < 0.05. The RR of minor bleeding for LMWH vs. UFH was 0.76 (95% CI: 0.64-0.92), p < 0.05. The RR of minor bleeding for LMWH vs. warfarin was 3.28 (95% CI: 2.21-4.70), p < 0.05. CONCLUSION: in orthopaedic surgery, LMWH is significantly superior to both UFH and warfarin in the prevention of DVT and results in significantly less minor bleeding complications when compared to UFH, but significantly more minor bleeding when compared to warfarin. PMID- 9212356 TI - Effect of dermatan sulphate on activated partial thromboplastin time determined with different reagents. AB - Five widely used activated partial thromboplastin time (APTT) reagents (Actin-FS, Actin-FSL, Hemolab Silimat, IL-Test APTT Ellagic Acid and Thrombofax Activated) were compared for their sensitivity and precision in measuring the effect of dermatan sulphate on blood coagulation. On each of 4 days, aliquots of the same normal human plasma pool were mixed with dermatan sulphate (MF701) at concentrations ranging from 10 to 100 micrograms/ml, and APTT was measured in duplicate with all reagents by a photo-optical coagulometer. The order of testing between and within reagents was changed every day. The relationship of APTT ratio to dermatan sulphate concentration was linear with all the reagents. There were statistically significant differences between reagents in their sensitivity to DS, as reflected by linear regression slopes. IL-Test was the most sensitive reagent. At dermatan sulphate concentrations of 20, 50, and 80 micrograms/ml APTT ratio ranged from 1.5 to 1.7, 1.9 to 2.3 and 2.3 to 2.9, respectively, according to the reagent. The lambda coefficient and coefficient of variation derived from regression analysis, both reflecting assay precision, ranged from 0.57 to 0.71 and from 4.6 to 5.1%, respectively, with all but the least precise reagent. The best sensitivity/precision balance was displayed by IL-Test. The APTT reagent should therefore be standardized, with special regard to sensitivity to DS, when testing the relationship of dermatan sulphate clinical effects to APTT response. PMID- 9212357 TI - Platelet count and the risk of bleeding in hospitalized patients with venous thromboembolism starting anticoagulant therapy. AB - In a series of patients with pulmonary embolism (PE) we have previously demonstrated that the risk of recurrent PE was inversely correlated to platelet count (PlC) levels. To find out whether PlC levels were also associated to a different incidence of heparin-related bleeding complications, we report our experience with 1,103 consecutive patients with venous thromboembolism (VTE) receiving full-dose heparin therapy. Six points of clinical and laboratory information were recorded on admission and then compared to the development of bleeding: the patient's age and sex; the etiology of VTE; the type of heparin used (unfractioned, UFH, vs. low-molecular-weight, LMWH), the presence or lack of PE findings on lung scan, and the PC levels on admission. Bleeding occurred in 64/1,103 patients (6%). Patients who bled were significantly older than those who did not (72 +/- 11 vs. 64 +/- 17 years; p = 0.0005). There were no significant differences in bleeding rate according to any of the risk factors that could have predisposed to VTE, but patients treated with UFH bled significantly more frequently than those on LMWH (48/636 vs. 16/467; odds ratio: 2.30; 95% confidence interval: 1.25-4.28). Finally, mean PlC levels were significantly lower at VTE diagnosis in patients who subsequently bled (227 +/- 112 vs. 262 +/- 110 x 10(9) liters-1; p = 0.01). The logistic regression analysis confirmed that all three variables were independent risk factors for bleeding complications. This is the first study to demonstrate that PlC levels (within the normal range) are inversely correlated with the risk of heparin-related bleeding. These findings may be interest not only from the point of view of pathogenesis but also clinically, as they may be used in the decision as to which VTE patients could receive heparin therapy at home. PMID- 9212358 TI - Prediction of the transfusion effect of platelet concentrates as measured by a model of primary hemostasis ex vivo. AB - A method to determine primary hemostasis ex vivo (Thrombostat) was modified to monitor the transfusion effect of platelet concentrates (PC) in 12 patients with thrombocytopenia following bone marrow transplantation. It was possible to measure platelet function in patients with a platelet count lower than 2 x 10(10)/l. In addition, the platelet aggregometer (Born) was adapted to determine cell function in PC anticoagulated with acid citrate dextrose of citrate phosphate dextrose. It was possible to make a prediction (r = 0.89) of the effect of a given PC on a patient's ex vivo primary hemostasis parameters. Platelet aggregation following addition of 20 muM ADP to PC, obtained from 12 single donors, resulted in an average maximal light transmission (light transmission/age of concentrate in days) of 61%/1 day and 37%/5 days, respectively. The same experiment gave only 39%/1 day and 13%/4 days for pooled platelets. To avoid possible immunization and bleeding complications, a reliable monitoring of platelet transfusion seems highly desirable. PMID- 9212359 TI - C-reactive protein: structural biology, gene expression, and host defense function. AB - Over the past few years substantial insight was gained into the biology and biochemistry of human C-reactive protein (CRP). X-ray crystallography in conjunction with mutational analyses, generated the three-dimensional structure of the protein and indicated the topology and structure of ligand-binding sites. Using human CRP transgenic mice infected with Streptococcus pneumoniae, we obtained data that clearly established CRP as an important host defense molecule. Studies using the same mice revealed a previously unknown testosterone-dependence of constitutive expression of human CRP. In this article we provide a brief overview of these recent findings. PMID- 9212360 TI - Temporal discontinuities in progression of NOD autoimmune diabetes. AB - Consideration of the pathophysiology of insulin-dependent diabetes mellitus in the nonobese diabetic (NOD) mouse can be viewed from a temporal perspective. We argue that there are discontinuous phases and each phase may reflect a phenotype educed by a particular set of genetic and epigenetic events. Therefore, temporal dissection may be a useful platform for causal dissection and we have set out this article as follows: 1. Introduction. 2. "Pre-time." a. Genetics. b. Parental effects. 3. Development of insulitis. a. Development of autoimmunity vs waning of or failure to establish tolerance. b. Importance of beta cell mass. c. Homing. 4. Onset of beta cell destruction. 5. Further Discussion. PMID- 9212361 TI - Bcl-x and the regulation of survival in the immune system. AB - A variety of experimental models indicate that programmed cell death, or apoptosis, of lymphocytes is a key mechanism in the homeostatic regulation of immunity. Apoptosis is important in early B- and T-cell development to delete cells with nonfunctional antigen receptors, and is also critical for censoring self-reactive cells at the immature lymphocyte stage and at various stages after lymphocytes reach maturity. In this article we focus on the role of the apoptosis regulatory gene bcl-x in controlling survival during lymphocyte development and following B- and T-cell activation. Interesting parallels are observed for bcl-x expression between the B- and T-lineages. The available data also indicate that bcl-x and bcl-2 are expressed in reciprocal patterns during the lifespan of a lymphocyte, suggesting unique regulatory roles for these two survival proteins. PMID- 9212362 TI - Differentiation and apoptosis of human germinal center B-lymphocytes. AB - An in vitro experimental model was developed to characterize the cellular and molecular factors that regulate germinal center (GC)-B-cell differentiation and apoptosis. In the culture system that sustains the GC-B-cell survival, CD40L stimulation is essential for GC-B-cell proliferation and differentiation in the presence of 1L-2, IL-4, and IL-10. IL-2 and Il-4 promote proliferation of GC-B cells, whereas IL-10 is required for generation of plasma cells. Generation of memory B cells requires CD40L, IL-2, IL-4, but not IL-10. There are two mechanisms that cause apoptosis. In the early stage, spontaneous apoptosis occurs in the absence of CD40 stimulation. Following CD40L stimulation, Fas-mediated apoptosis operates to eliminate GC-B-cells, unless activated GC-B-cells encounter a second signal via B-cell Ig receptors. Physiological significance of these findings is discussed. PMID- 9212363 TI - The CD94/NKG2 C-type lectin receptor complex: involvement in NK cell-mediated recognition of HLA class I molecules. AB - A multigene family to human Ig-SF receptors and members of the murine Ly49 C-type lectin family are involved in natural killer (NK) cell-mediated recognition of MHC class I molecules. The human CD94 glycoprotein covalently assembles with different C-type lectins of the NKG2 family. By functional criteria, the CD94/NKG2-A (kp43) receptor complex appears also involved in NK cell-mediated recognition of different HLA class I allotypes. Similarly to the other NK inhibitory receptors, NKG2-A contains cytoplasmic immunoreceptor tyrosine-based inhibitory motifs (ITIMS). By contrast, NK clones bearing different receptor complex (CD94/p39) are triggered upon ligation by CD94-specific monoclonal antibodies (MAbs); the p39 subunit is likely encoded by other member(s) of the NKG2 family. Expression of different CD94/NKG2 complexes is warranted to precisely assess their specific interaction with HLA class I molecules, and the molecular basis for their divergent functional properties. PMID- 9212366 TI - [Susceptibilities of bacteria isolated from patients with respiratory infectious diseases to antibiotics (1995)]. AB - The bacteria isolated from the patients with lower respiratory tract infections were collected by institutions located throughout Japan, since 1981. Ikemoto et al. have been investigating susceptibilities of these isolates to various antibacterial agents and antibiotics, and characteristics of the patients and isolates from them each year. Results obtained from these investigations are discussed. In 23 institutions around the entire Japan, 567 strains of presumably etiological bacteria were isolated mainly from the sputa of 459 patients with lower respiratory tract infections during the period from October 1995 to September 1996. MICs of various antibacterial agents and antibiotics were determined against 74 strains of Staphylococcus aureus, 82 strains of Streptococcus pneumoniae, 104 strains of Haemophilus influenzae, 85 strains of Pseudomonas aeruginosa (non-mucoid strains), 18 strains of Pseudomonas aeruginosa (mucoid strains), 52 strains of Moraxella subgenus Branhamella catarrhalis, 25 strains of Klebsiella pneumoniae etc., and the drug susceptibilities of these strains were assessed except for those strains that died during transportation. 1) S. aureus. S. aureus strains for which MICs of oxacillin (MPIPC) were higher than 4 micrograms/ml (methicillin-resistant S. aureus) accounted for 52.7%. Arbekacin (ABK) showed the most highest activity against S. aureus with MIC80 of 0.5 micrograms/ml. Vancomycin (VCM) showed the next highest activity with MIC80 of 1 microgram/ml. These drugs showed the high activities against MRSA with MIC80S of 1 microgram/ml. 2) S. pneumoniae. Most of drugs tested showed potent activities against S. pneumoniae. Imipenem (IPM) and panipenem (PAPM), carbapenems, showed the most potent activity with MIC80S of 0.063 microgram/ml. Cefotaxime (CTX), cefmenoxime (CMX) and cefpirome (CPR) of cephems showed the next most potent activities with MIC80S of 0.25 microgram/ml. Erythromycin (EM) and clindamycin (CLDM) showed low activities with MIC80S 128 micrograms/ml or high. Among these strains, however, 48.8% and 65.9% of respective strains were quite toward sensitive these agents with MICs of 0.063 microgram/ml. 3) H. influenzae. The activities of all drugs were potent against H. influenzae test with all MICs at 4 micrograms/ml or below. Cefotiam (CTM), CMX, cefditoren (CDTR) and ofloxacin (OFLX) showed the most potent activity with MIC90S to 0.063 microgram/ml. 4) P. aeruginosa. (mucoid strains) IPM and tobramycin (TOB) showed the most potent activity against P. aeruginosa (mucoid strains) with MIC80S of 1 microgram/ml. Ceftazidime (CAZ), cefsulodin (CFS) and carumonam (CRMN) showed next potent activity, with MIC80S of 2 micrograms/ml. The MIC80S of the other drugs ranged from 4 micrograms/ml to 32 micrograms/ml. 5) P. aeruginosa (non mucoid strains). TOB and ciprofloxacin (CPFX) showed the most potent activities against P. aeruginosa (non-mucoid strains) with MIC80S of 1 microgram/ml. The MIC80 of ampicillin (ABPC) was 128 micrograms/ml in 1994, it was 16 micrograms/ml in 1995. 6) K. pneumoniae. All drugs except ABPC were active against K. pneumoniae. CPR and CRMN showed the most potent activities against K. pneumoniae with MIC80S of 0.063 microgram/ml. The MIC80S of the other drugs ranged from 0.125 microgram/ml to 2 micrograms/ml. 7) M. (B.) catarrhalis. Against M. (B.) catarrhalis, all the drugs showed good activities with MIC80S at 4 micrograms/ml or below. And MICs of all strains were 8 micrograms/ml or below. IPM, OFLX and minocycline (MINO) showed the most potent activity with MIC80S of 0.063 microgram/ml. Also, we investigated year to year changes in the characteristics of patients, their respiratory infectious diseases, and the etiology. Patients' backgrounds were examine for 567 isolates from 459 cases. The examination of age distribution found that the proportion of patients with ages over 60 years was 66.3% of all the patients showing a slight increase over that in 1994. Proportion of differe PMID- 9212365 TI - HLA-C revisited. Ten years of change. AB - During the past 10 years knowledge about the interactions between major histocompatibility complex (MHC) class I molecules and the T-cell receptor (TCR) complex of cytotoxic T-cells (CTL) has developed dramatically. But the primary interest, both with respect to structure as well as function, has concentrated on HLA-A and -B molecules because of their high sequence polymorphism and their dominating presence at the cell surface. In contrast, HLA-C molecules seemed to be of only minor importance in the cascade of immune reactions owing to their more limited polymorphism and reduced levels of surface expression. The inability to define a number of antigen specificities had the result that HLA-C molecules were often neglected in studies of immune response, transplantation, and disease association. More recently a new function has been identified for HLA class I molecules where they act as inhibitors of the lytic capacity of natural killer (NK) cells and non-MHC-restricted T-cells. Moreover, the understanding of this novel mode of negative regulation of cytotoxicity was remarkably influenced by HLA-C since these were the first HLA class I molecules found to have such inhibitory potential. With this new inhibitory function serving as an essential component of the immune system, HLA-C molecules can no longer be neglected. PMID- 9212368 TI - [Intravaginal bacterial flora and clinical significance of granulocyte elastase and pH determination]. AB - In 95 patients with abnormal vaginal and cervical secreta (49 pregnant women and 46 non-pregnant women), the relation between intravaginal flora and intravaginal granulocyte elastase (Elastase) and pH was investigated. The results were as follows. 1) Gram-positive bacteria were detected in the vaginal secreta at a high rate (87/144, 60.4%), and it mainly consisted of Lactobacillus sp. (67/142, 46.5%). It was followed anaerobia (26/144, 18.1%) and fungi (26/144, 18.1%). 2) The patients with cervicitis or vaginitis had higher elastase value (6.65-6.69 micrograms/ml) than the ones with vaginal erosion, and the patients who showed an intravaginal pH value not lower than 5.0 had significantly increased elastase value (6.44 +/- 1.40 micrograms/ml) than the patients who showed the values 4.5 or higher. 3) Regarding the relation between the detected bacteria and elastase values, elastase values were higher in the patients infected by anaerobia (6.58 +/- 1.40 micrograms/ml), Gram-negative bacteria (6.01 +/- 3.61 micrograms/ml), Gram-positive bacteria (5.02 +/- 0.94 micrograms/ml) and fungi (5.14 +/- 1.08 micrograms/ml) than the values in patients with Lactobacillus sp. (pH < 4.5). Further, the intravaginal pH value was higher than 4.5 in all of these groups, which was higher compared with the one in the patients infected with Lactobacillus sp. (4.04 +/- 0.04). PMID- 9212364 TI - Nasal lymphoid tissue, intranasal immunization, and compartmentalization of the common mucosal immune system. AB - Mucosal application of vaccines with an appropriate adjuvant can induce immune responses at both systemic and mucosal sites, and therefore may prevent not only infectious disease, but also colonization of mucosal surfaces. Intranasal is more effective than intragastric immunization at generating earlier and stronger mucosal immune response. Nasal lymphoid tissue (NALT) and its local draining lymph nodes may retain long-term immune memory. IgA isotype switching, and the differentiation and maturation of IgA antibody-secreting cells (ASC) may occur before these cells migrate out of NALT, whereas IgG ASC responses require passage of the cells through draining lymph nodes of the NALT. Knowledge of whether immune memory cells can recirculate to and reside in the inductive sites other than their origin after encountering antigen will be helpful for understanding the compartmentalization of the common mucosal immune system as well as for determining the best route for delivering a mucosal vaccine against a particular pathogen. PMID- 9212367 TI - [Isolation rate of Enterococcus spp. from surgical infections and their susceptibilities]. AB - Enterococcus spp. isolated from surgical infections during the period from July 1982 to June 1995 were investigated in a multicenter study involving 19 hospitals in Japan, and the following results were obtained. 1. Though the isolation rate of Enterococcus faecalis and other Enterococcus spp. were not high from primary infections, and from postoperative infections the isolation rate of other Enterococcus spp. was also low, the isolation rate of E. faecalis was highest from postoperative infections after 1993. 2. Vancomycin (VCM) showed strongest activity against E. faecalis, and followed by those of ampicillin (ABPC), imipenem. levofloxacin (LVFX) and meropenem in this order. Against other Enterococcus spp., VCM showed strongest activity, and followed by those of ABPC and LVFX. There were no resistant strains against VCM. PMID- 9212369 TI - [Measurement of antibody titers to chlamydial infection and effects of levofloxacin in cystic cervicitis and chlamydial infection]. AB - Generally, clinical symptoms such as abnormal leukorrhea are caused by C. trachomatis, an ordinary bacteria in cervical infection. The effects of levofloxacin administration at a dose of 300 mg/day for 5-14 days were investigated in the subjects (n = 66) after the discussion of cystic cervicitis. The treatment was made in combination with chloramphenicol-solcoseryl tablet for vaginal use. And it was demonstrated that treatment was effective in all subjects. Then, Sero IPALISA, an examination of IgA/IgG antibody was conducted for the screening of Chlamydia infection (n = 258). The rate of antibody-positive case was 48/160 (30.0%) for the non-pregnant women and 26/98 (26.5%) for the pregnant. The occurrence rates for the women singing in 15-24 and 35-39 years of age were 50 and 41%, respectively. The results from the measurement of the antibody titer were as follows: the rate of IgA/IgG positive care was 61/87 for IgA and 56/87 for IgG when the cut-off index was 1.11 or more. The rates for both antibody were 11/87 (12.6%) and 24/87 (27.6%) for the indexes of 1.11-3.00 and 3.01 or more, respectively. Next, one to three courses levofloxacin at 300 mg/day for 14 days were given to 48 non-pregnant subjects infected with Chlamydia and one to two courses of clarithromycin at 400 mg/day for 14 days were given to 26 pregnant subjects. Side effects have not been noted in any care and there was no changes in the IgA/IgG antibody titer depending on these treatments. PMID- 9212370 TI - Diagnosis and assessment of depression and suicidality using the NIMH Diagnostic Interview Schedule for Children (DISC-2.3). AB - The diagnostic Interview Schedule for Children (DISC-2.3) was studied in a sample of 265 adolescent inpatients to determine type and concurrent validity of depressive symptoms and depressive disorder diagnoses for different DISC-2.3 informants (parent, adolescent, both). The Children's Depression Rating Scale- Revised, Reynolds Adolescent Depression Scale (RADS), Suicide Ideation Questionnaire--Junior, Spectrum of Suicide Behavior Scale, and clinical consensus diagnoses were used to assess concurrent validity. Results indicated that (1) parents, compared to adolescents, reported a higher prevalence of all depressive symptoms with the exception of weight change; (2) DISC-2.3 depressive and suicidality symptoms were related positively to independent validating criteria for all informant conditions, suggesting good concurrent validity; (3) the DISC 2.3 both informant condition correctly identified the most depressive disorders; and (4) the parent, but not the adolescent, DISC-2.3 Informant condition contributed to the prediction of clinical consensus diagnoses of depression after taking into account RADS scores. PMID- 9212372 TI - School-based secondary prevention for children with disruptive behavior: initial outcomes. AB - First through fourth graders from 22 suburban elementary schools were screened for cross-setting disruptive behavior as eligibility criteria for participation in a longitudinal secondary prevention study aimed at reducing the risk for serious externalizing behavioral disorders. Three hundred nine subjects participated in either a multicomponent competence enhancement intervention (MCEI) or an information/attention control (IAC) condition over a 2-year period. Following baseline requirements, initial intervention effects were assessed at the end of intervention Year 1, at the beginning of intervention Year 2 (fall of the next school year), and at the end of intervention Year 2. Multisource assessments were not supportive of the efficacy of the MCEI over the IAC condition. Children in both groups rated themselves as improved over time in terms of increased adaptive skills and decreased school problems and internalizing symptoms. Teacher and parent ratings of externalizing behavior did not yield evidence of positive change, but teachers noted improved problem solving and observers noted a decrease in behavioral interference in both groups over time, possibly as a result of maturation. PMID- 9212371 TI - Behavioral and emotional problems in young preschoolers: cross-cultural testing of the validity of the Child Behavior Checklist/2-3. AB - The cross-cultural validity of the Child Behavior Checklist for Ages 2-3 (CBCL/2 3) was tested in three Dutch samples of children referred to mental health services, from the general population, and from a twin study. Six scales were derived from factor analyses and labeled Oppositional, Aggressive, and Overactive, which constituted a broadband Externalizing grouping; Withdrawn/Depressed and Anxious, which constituted a broadband Internalizing grouping; and Sleep Problems. Internal consistencies of the scales, their test retest reliabilities, interparent agreement, discriminative power, predictive relations with problem ratings 2 years later, and relations to other instruments designed to measure general development and behavior problems were adequate, and highly comparable to psychometric properties in American samples. It was concluded that across languages and cultures behavioral/emotional problems of young preschoolers may be adequately assessed with the CBCL/2-3. PMID- 9212373 TI - Levels of analysis in cognitive bases of maternal disciplinary dysfunction. AB - This study tested alternative hypotheses concerning relations between mothers' disciplinary dysfunction and their descriptive versus inference-level interpretations of child noncompliance. Mothers of aggressive boys (MAGGs; n = 19) and mothers of average boys (MAVGs; n = 17) were presented with hypothetical vignettes of compliance situations (mean ages: mothers = 26.8 years, children = 4.5 years). Each vignette ended with the child being compliant or with each of a variety of noncompliant behaviors (request, statement, compliant, ignore, or oppose). Dependent variables were mothers' judgments of noncompliance severity (a descriptive measure), and attributions of defiant intent to the child (an inferential measure). Findings across analyses consistently pointed to attributions as more discriminating than judgments in differentiating between maternal groups. It was concluded that models of maternal discipline dysfunction should focus on analysis of inferential rather than descriptive cognitive responses to child noncompliance, and that parenting interventions should incorporate attribution-training into treatment protocols. PMID- 9212374 TI - Sex differences in referral rates of children with gender identity disorder: some hypotheses. AB - From 1978 through 1995, a sex ratio of 6.6:1 of boys to girls (N = 275) was observed for children referred to a specificity clinic for gender identity disorder. This article attempts to evaluate several hypotheses regarding the marked sex disparity in referral rates. The sexes did not differ on four demographic variables (age at referral, IQ, and parent's social class and marital status) and on five indices of general behavior problems on the Child Behavior Checklist; in addition, there was only equivocal evidence that boys with gender identity disorder had significantly poorer peer relations than girls with gender identity disorder. Although the percentage of boys and girls who met the complete DSM-III-R criteria for gender identity disorder was comparable, other measures of sex-typed behavior showed that the girls had more extreme cross-gender behavior than the boys. Coupled with external evidence that cross-gender behavior is less tolerated in boys than in girls by both peers and adults, it is concluded that social factors partly account for the sex difference in referral rates. Girls appear to require a higher threshold than boys for cross-gender behavior before they are referred for clinical assessment. PMID- 9212375 TI - Neuromotor functioning and behavior problems in children at risk for psychopathology. AB - Previous studies have found that early neuromotor deficits may be a precursor of later psychopathology. The present study examined the relationship between neuromotor dysfunction and behavioral deviance in children characterized by a variety of risk factors (parental schizophrenia, parental psychiatric disorder other than schizophrenia, and parental maltreatment). The sample consisted of 108 children (average age 9.75 years) who were assessed twice, approximately 1 year apart. It was was found that maltreated children had poorer neuromotor functioning and more behavior problems than children who were not maltreated, regardless of parental psychiatric status. The results also indicated that the relationship between neuromotor functioning and problem behaviors varied as a function of parental psychiatric status. These findings suggest that, although the effects of maltreatment are generalized and pervasive, there are distinctive relationships between neuromotor functioning and behavioral deviance depending on the nature of the risk factors a child has been exposed to. PMID- 9212376 TI - Dysphoria and children's processing of supportive interactions. AB - This study examined the processing of supportive interactions by dysphoric and nondysphoric preteens and early adolescents. Seventy-two youngsters between the ages of 10 and 13 evaluated the supportiveness and helpfulness of standardized, videotaped interactions between a distressed preadolescent and a maternal figure. The tape presentations varied in terms of the level of depicted maternal support and instructional condition (degree of self-reference). The results indicated that dysphoric youngsters evaluated both the supportiveness and helpfulness of interactions less positively than nondysphoric agemates. Group differences in support evaluations were most pronounced in the self-referenced condition. The level of depicted support did not affect processing differences. Dysphoric subjects reported lower levels of emotional support in prior relationships and a greater tendency to view supportive behavior as ingenuine than nondysphoric peers. Variation in prior support experiences accounted for group differences in the evaluation of the supportiveness of new interactions. PMID- 9212377 TI - The moderating effects of children's fear and activity level on relations between parenting practices and childhood symptomatology. AB - Parenting practices have been previously linked to childhood symptomatology. However, little consideration has been given to the potential effect of individual differences within the child on this relation. The current study assessed the moderating effects of children's activity level and fear on relations between parenting practices and childhood aggression and depressive symptoms using a sample of 64 fourth-, and fifth-grade boys. The findings showed that poorly monitored active boys and fearful boys who were exposed to harsh discipline exhibited high levels of aggression. Boys characterized by high fear who were exposed to harsh discipline or whose parents were extremely overinvolved showed elevated levels of depressive symptoms. These findings suggest that integrating children's individual differences with parenting models enhances our understanding of the etiology of childhood symptomatology. The intervention implications of such an integration are discussed. PMID- 9212378 TI - An exploration of the relation between hostility and disease. AB - Hostility has been studied mainly in relation to coronary heart disease (CHD). However, given the pathways linking hostility to CHD, it might be expected that hostility also relates to non-CHD. Therefore, the relation between the expression and the experience of hostility and various health outcomes was examined in a cross-sectional design. The data were collected among male patients with a myocardial infarction in the age range of 30-70 years (N = 279) and a population sample of men in the same age group (N = 2663). Based on checklist of the most frequent disorders, the subjects from the latter group were divided into subsamples according to their disease status. Three components of hostility, i.e., resentment, suspicion, and aggression, were measured by the Buss Durkee Hostility Inventory (Buss & Durkee, 1957). The overall finding was that all components of hostility were related to non-CHD disease but not to CHD. PMID- 9212379 TI - The impact of psychosocial features of employment status on emotional distress in chronic pain and healthy comparison samples. AB - This study examined the extent to which measures of psychosocial features of employment status predict emotional distress in chronic pain (n = 83) and healthy comparison (n = 88) samples. Participants completed measures of emotional distress, pain severity, psychosocial features of employment status, and demographic data. After controlling for length of current unemployment, number of pain sites, and level of current pain severity, psychosocial measures (structured and purposeful time use, perceived financial security, skill use, social support form formal sources) were significant predictors of emotional distress in the chronic pain sample. Similar results were obtained for the healthy comparison sample. Structured and purposeful time use emerged as the most significant individual predictor of emotional distress for both samples. Findings are discussed in terms of their potential implications for treating chronic pain patients and the need to develop multidimensional measures that assess features of employment status within chronic pain samples. PMID- 9212380 TI - The effects of medical evidence and pain intensity on medical student judgments of chronic pain patients. AB - This study examined symptom judgments made by medical students of hypothetical chronic low back pain patients. Eight vignettes were varied as to the pain intensity reported by the hypothetical patient (low vs. moderate vs. high vs. very high) and the availability of medical evidence supportive of the pain report (present vs. absent). Ninety-five subjects read vignettes and made judgments of patient emotional distress, pain intensity, and pain-related disability. Subjects significantly discounted pain level when intensity was high but slightly augmented pain level when intensity was low. Judgments of pain and disability were higher for patients for whom medical evidence was present compared to those for whom it was absent. The results support and extend previous research on the effects of situational and patient variables on observer pain judgments. Future research should examine the influence of these biasing variables on the assessment and treatment of chronic pain patients. PMID- 9212381 TI - Illness self-schemas in depressed and nondepressed rheumatoid arthritis patients. AB - This study examined the hypothesized illness self-schemas construct in persons with rheumatoid arthritis (RA). Biases in self-description, information processing, and schema-consistent illness behavior were examined in depressed and nondepressed persons with RA and compared with those of depressed and nondepressed controls. Major findings revealed that RA-depressed subjects exhibited pervasively negative self-description and biased processing of negative illness-related information. RA-nondepressed subjects demonstrated a bias for positive self-description and enhanced processing of positive illness-related information. Using regression analysis, the illness self-schema construct predicted unique variance in self-reported functional disability. Findings are reviewed in the context of previous research on self-schemas, chronic pain, and cognitive variables in chronic illness. Potential clinical implications and directions for future research are discussed. The illness self-schema construct has significant heuristic value which could guide further research on the psychosocial adjustment of individuals with chronic illnesses. PMID- 9212382 TI - The assessment of diabetes-related cognitive and social factors: the Multidimensional Diabetes Questionnaire. AB - The purpose of this study was to examine the psychometric properties of the recently developed Multidimensional Diabetes Questionnaire (MDQ). The MDQ, which is theoretically linked to a social learning perspective of diabetes, was designed to provide a comprehensive assessment of diabetes-related cognitive and social factors. It includes 41 items grouped into three sections: (1) perceptions related to diabetes and related social support, (2) positive and misguided reinforcing behaviors related to self-care activities, and (3) self-efficacy and outcome expectancies. Confirmatory factor analyses, conducted on a sample of 249 patients with non-insulin-dependent diabetes mellitus, supported the construct validity of the MDQ. Adequate internal consistency and significant demographic, psychological, behavioral, and disease-related correlates were found. The MDQ may prove valuable in understanding individual differences in adjustment to diabetes. PMID- 9212384 TI - Effect of electrochemically deposited apatite coating on bonding of bone to the HA-G-Ti composite and titanium. AB - The surfaces of hydroxyapatite-glass-titanium (HA-G-Ti) functionally gradient composite and titanium bars were treated with electrochemical apatite deposition, and a cathodic current was applied at 62 degrees C in a solution containing calcium and phosphate ions. Specimens with and without the electrochemical surface treatment were implanted in the femurs of Japanese white rabbits. The rabbits were sacrificed at 3, 6, and 9 weeks after implantation, and the bonding strengths of bone to these specimens were determined by a pull-out method. At 3 and 6 weeks after implantation the specimens with the electrochemical surface treatment showed larger values for the Weibull modulus and characteristic strengths than those of untreated specimens, whereas there was no remarkable difference in the results at 9 weeks. Especially the pull-out strengths of surface-treated specimens were significantly larger than the untreated ones at 3 weeks after implantation. Scanning electron microscopy and Fourier transform infrared absorption spectroscopy of the specimen surface after implantation demonstrated that formation of new bone was enhanced by the electrochemical surface treatment. It can be concluded that the electrochemical surface treatment undoubtedly contributes to the early stage fixation between bone and implant. PMID- 9212383 TI - Ectopic bone formation by marrow stromal osteoblast transplantation using poly(DL lactic-co-glycolic acid) foams implanted into the rat mesentery. AB - Porous biodegradable poly(DL-lactic-co-glycolic acid) foams were seeded with rat marrow stromal cells and implanted into the rat mesentery to investigate in vivo bone formation at an ectopic site. Cells were seeded at a density of 6.83 x 10(5) cells/cm2 onto polymer foams having pore sizes ranging from either 150 to 300 to 710 microns and cultured for 7 days in vitro prior to implantation. The polymer/cell constructs were harvested after 1, 7, 28, or 49 days in vivo and processed for histology and gel permeation chromatography. Visual observation of hematoxylin and eosin-stained sections and von Kossa-stained sections revealed the formation of mineralized bonelike tissue in the constructs within 7 days postimplantation. Ingrowth of vascular tissue was also found adjacent to the islands of bone, supplying the necessary metabolic requirements to the newly formed tissue. Mineralization and bone tissue formation were investigated by histomorphometry. The average penetration depth of mineralized tissue in the construct ranged from 190 +/- 50 microns for foams with 500-710-microns pores to 370 +/- 160 microns for foams with 150-300-microns pores after 49 days in vivo. The mineralized bone volume per surface area and total bone volume per surface area had maximal values of 0.28 +/- 0.21 mm (500-710-microns pore size, day 28) and 0.038 +/- 0.024 mm (150-300-microns, day 28), respectively. As much as 11% of the foam volume penetrated by bone tissue was filled with mineralized tissue. No significant trends over time were observed for any of the measured values (penetration depth, bone volume/surface area, or percent mineralized bone volume). These results suggest the feasibility of bone formation by osteoblast transplantation in an orthotopic site where not only bone formation from transplanted cells but also ingrowth from adjacent bone may occur. PMID- 9212385 TI - Bone formation by three-dimensional stromal osteoblast culture in biodegradable polymer scaffolds. AB - Bone formation was investigated in vitro by culturing stromal osteoblasts in three-dimensional (3-D), biodegradable poly(DL-lactic-co-glycolic acid) foams. Three polymer foam pore sizes, ranging from 150-300, 300-500, and 500-710 microns, and two different cell seeding densities, 6.83 x 10(5) cells/cm2 and 22.1 x 10(5) cells/cm2, were examined over a 56-day culture period. The polymer foams supported the proliferation of seeded osteoblasts as well as their differentiated function, as demonstrated by high alkaline phosphatase activity and deposition of a mineralized matrix by the cells. Cell number, alkaline phosphatase activity, and mineral deposition increased significantly over time for all the polymer foams. Osteoblast foam constructs created by seeding 6.83 x 10(5) cells/cm2 on foams with 300-500 microns pores resulted in a cell density of 4.63 x 10(5) cells/cm2 after 1 day in culture; they had alkaline phosphatase activities of 4.28 x 10(-7) and 2.91 x 10(-6) mumol/cell/min on Days 7 and 28, respectively; and they had a cell density that increased to 18.7 x 10(5) cells/cm2 by Day 56. For the same constructs, the mineralized matrix reached a maximum penetration depth of 240 microns from the top surface of the foam and a value of 0.083 mm for mineralized tissue volume per unit of cross sectional area. Seeding density was an important parameter for the constructs, but pore size over the range tested did not affect cell proliferation or function. This study suggests the feasibility of using poly(alpha-hydroxy ester) foams as scaffolding materials for the transplantation of autogenous osteoblasts to regenerate bone tissue. PMID- 9212386 TI - Plasma surface modification of artificial corneas for optimal epithelialization. AB - We have demonstrated that the optimal surface treatment of a polyvinylalcoholcopolymer hydrogel for epithelial cell migration and proliferation is an argon radio frequency (rf) plasma treatment. The surface chemistry of the material was determined prior to each cellular evaluation, allowing us to compare the biological response with a known surface chemistry. The cellular response was carried out in a consistent manner a minimum of three separate runs. We found that the optimal conditions required culturing the cells under constant rotation. Cells became confluent on argon-plasma-treated surfaces coated under several different reactions pressures, and after 2 weeks they became multilayered. Our experiments demonstrated that cells proliferated and extracellular matrix and adhesion proteins were present only when the surface was treated with an argon rf plasma; acetone- and ammonia-treated surfaces did not yield the desired results. Organ culture experiments further demonstrated the efficacy of the argon-treated surfaces. In these experiments, intact keratoprosthetic devices with modified hydrogel surfaces were implanted into rabbit corneas. The excised corneas containing the devices were cultured, and 3 weeks later, using confocal laser scanning microscopy, confluent epithelium was detected on the modified hydrogel surface. This is the first demonstration that rabbit limbal epithelial cells can migrate onto a synthetic cornea containing a modified hydrogel-treated surface and form a confluent surface of epithelium. PMID- 9212387 TI - Intramedullary implant of plasma-sprayed hydroxyapatite coating: an interface study. AB - An intramedullary implant model in the canine femora was developed to evaluate the mechanical and histological responses between cancellous bone and plasma sprayed hydroxyapatite coatings (HACs) on ti-6A1-4V implants, with 12- and 24 week follow-ups. HACs of different thicknesses were investigated. Results of the mechanical testings revealed that after 24 weeks of implantation, the mean shear strength (2.49 +/- 0.12 MPa) of the 50 microns HACs was significantly higher (p < 0.05) than that of the 200 microns HACs (1.44 +/- 0.19 MPa). However, using backscattered electron images (BEIs) throughout all the implant periods, no substantial histological variations in the extent of new bone apposition between the two HACs were observed. Occasionally, solution-mediated disintegration of the 50 microns HAC was found 24 weeks postimplantation. Histomorphometric studies from the BEIs demonstrated that for both HACs the percentage of the direct HAC cancellous bone contact was approximately 50% at 12 weeks and 75% at 24 weeks. After the mechanical tests, the 200 microns HACs had fracture sites either inside the coating layers or at the HAC-titanium interfaces, which might explain why the mechanical performance of the 200 microns HACs was inferior to that of the 50 microns HACs even though both HACs had the same histological behaviors. PMID- 9212388 TI - Influence of ePTFE polymer implant permeability on the rate and density of corneal extracellular matrix synthesis. AB - Microporous polymers have great potential for the production of corneal keratoprosthetic devices. Keratocytes invade the pores of expanded polytetrafluoroethylene implants (ePTFE) and collagen synthesis occurs. This ePTFE becomes translucent after its implantation in the stroma of rabbit cornea. The rate and density of cell growth within this polymer depends on the implant thickness, pore size, and its placement in the cornea. We have investigated the influence of the polymer permeability on the collagen and protein contents ePTFE implants. Rabbit corneal stroma were implanted with ePTFE disks (6 mm in diameter) by intralamellar keratoplasty. The implanted polymers were removed from the stroma after 3 to 6 months. The collagen and protein contents were determined after pepsin solubilization. The collagen content of the high-permeability implant was 3.7-fold greater than that of the low-permeability implant 3 months after implantation and 2.4-fold greater after 6 months. The total protein content of the high-permeability implant was 2.5-fold greater than that of low permeability implant at 3 months and was the same after 6 months. The collagen-to protein ratio was 68% in the high-permeability implants, and thus similar to that of normal corneal stroma. Thus, high polymer permeability increased both the rate and density of the corneal extracellular matrix ingrowth. PMID- 9212389 TI - Healing of gaps around calcium phosphate-coated implants in trabecular bone of the goat. AB - Hydroxylapatite coatings are under clinical investigation in orthopaedics and dentistry. Bone formation on apatite coatings in the presence of gaps is important for clinical applications. The importance of the stability of the coating is not known at present. By varying the plasma-spray parameters, and by the addition of fluoride, the crystallinity and stability of calcium phosphates can be changed. It is suggested that bone formation is enhanced by dissolution of the apatite coating. We studied apatite coatings of varying stability with regard to their gap-healing characteristics, and we examined what the maximum gap would be that can be bridged if a coating is applied. Ti-6A1-4V implants coated with 62% crystalline hydroxylapatite, 30% crystalline hydroxylapatite or fluorapatite, or noncoated Ti-6A1-4V were implanted in 16 goats. The implants were surrounded by gaps of 1 or 2 mm, and the follow-up period was 6 weeks. Histological examination and histometry revealed that gaps of 1 mm can be bridged by bone if an apatite coating is applied. However, only a minimal amount of bone contact was seen on the apatite coatings with 2 mm gaps. Uncoated implants demonstrated no bone contact at all. Among the three different coatings there were no differences in gap healing. It can be concluded that in the goat, gaps of 2 or more mm between coated implants and host bone tissue inhibit bone deposition on the coating (p < 0.05), but the stability of the coating does not influence gap healing characteristics. PMID- 9212390 TI - Development of degradable polyesterurethanes for medical applications: in vitro and in vivo evaluations. AB - To evaluate the biocompatibility of a newly developed degradable class of polyesterurethanes and their possible use as biomaterials, we investigated the cell and tissue interactions with these polymers using a small number of chemical base entities. The polymers were prepared by chain extension with diisocyanates of PHB/HV-diol and either PCL-diol or Diorez, another aliphatic polyester-diol. Regardless of the chemical composition of the four tested polyesterurethanes used as substrates, no morphological difference was observed either in the macrophages (macrophage cell line J774) or in the fibroblasts (fibroblast cell line 3T3) cultured on the polymers. In contrast, however, cell adhesion and growth of macrophages and fibroblasts were affected by the polymer properties. Compared to macrophages cultured on tissue culture polystyrene (TCPS), cells cultured on the test polymers exhibited levels of cell adhesion that varied from 65-100% of TCPS, and the doubling time was 25-43% higher on the polymers than on TCPS. Likewise, fibroblasts adhered to the polymers at lower rates (50-85% of TCPS) and grew at higher doubling times (125-140% of TCPS). Furthermore, cells cultured on the test polymers preserved their phenotypes: fibroblasts produced high amounts (up to 280% of control cells) of collagens Type I and Type IV and fibronectin; and macrophages produced nitric oxide (NO) and tumor necrosis factor alpha (TNF alpha) in the same concentrations as control cells and responded to lipopolysaccharide treatment by the elevation of the production of NO and TNF alpha, indicating that the cell-to-polymer interactions allow fibroblasts and macrophages to maintain their phenotypes. In vivo investigations showed that all four test polymers exhibit favorable tissue compatibility. The formed capsule was 60-250 microns thick. In addition, the polymers are degradable. After one year's subcutaneous implantation in rats, the molecular weight of the test polymers were reduced to about 50%, depending on the composition. Taken collectively, the present data demonstrate that the newly developed polyesterurethanes are cell and tissue compatible and biodegradable. PMID- 9212392 TI - Influence of substrate material and surface finishing on the morphology of the calcium-phosphate coating. AB - The formation of an apatite-like layer was achieved by immersing Ti-6A1-4V, Ti-Al 2.5Fe, and 316 L stainless-steel substrata in Hank's balanced salt solution (HBSS). The layer was characterized by surface analysis techniques, namely X-ray microanalysis and X-ray diffraction, and the morphology was observed by scanning electron microscopy and atomic force microscopy. The concentrations of Ca and P were monitored as a function of time. The morphology of the precipitate layer seems to be dependent both on the type of metal substrate and its surface finish. Polished Ti-6A1-4V and Ti-Al-2.5Fe surfaces exhibit a plate precipitate morphology, whereas rougher surfaces show scattered crystal-like precipitation. The results suggest that the layer produced by immersion of polished titanium alloys in HBSS is constituted by an amorphous apatite. PMID- 9212391 TI - Mechanical and histologic evaluation of Ca-P plasma-spray and magnetron sputter coated implants in trabecular bone of the goat. AB - To aim of this study was to investigate the bone response to calcium phosphate (Ca-P) plasma-spray and radiofrequency magnetron sputter-coated implants with comparable roughness. Therefore, tapered conical screw designed implants were installed in the trabecular bone of the femurs of nine goats. They were provided with two types of coatings, a plasma-spray dual coating of fluorapatite and hydroxyapatite (FA/HA-PS) and a titanium plasma-spray coating, covered with an amorphous Ca-P magnetron sputtercoating (TPS/Ca-P-a). These implants were evaluated histologically and mechanically after 3 months of implantation. A well controlled method to apply and measure a torsional force to load the screw-type implants to the point of failure was introduced. All implants healed uneventful and were well fixed. No significant difference (Student t test, p > 0.05) for the torsional failure force was measured for both type of coatings. Nevertheless, SEM revealed differently situated fracture planes. Light microscopy showed intimate bone-implanted contact for both types of coatings; original drill margins were still visible. A lamellar type of bone with some remodeling lacunae was shown. Histomorphometry revealed a higher percentage of bone contact for the FA/HA-PS coated implants (students t test, p < 0.05). Measurement of the amount of bone revealed more bone mass around TPS/Ca-P-a-coated implants (analysis of variance and Turkey multiple comparison, p < 0.05). PMID- 9212393 TI - Stereomorphologic observation of bone tissue response to hydroxyapatite using SEM with the EDTA-KOH method. AB - To obtain further information on the interaction of hydroxyapatite (HA) and the bony implantation bed, 20- to 40- mesh dense HA particles were implanted into the tibiae of dogs. Following healing periods of 2 weeks, 1 month, and 3 months, the specimens were retrieved and prepared by either conventional preparatory procedures for scanning electron microscopy (SEM) of the EDTA-KOH method. Under SEM observation, the interparticular osteogenesis among HA particles progressed in a programmed sequence. Ample blood supply and osteoblasts initially presented in the interparticular space. The secretion of bone matrix resulted in the formation of immature bone. This scaffold was then transformed into mature lamellar bone during the following bone remodeling process. The serial changes closely resembled the pattern viewed in controls that did not implant HA. A spatial relationship between bone cells and HA was clearly demonstrated. In particular, the osteoblasts displayed an extremely flat appearance with many microappendages. The microappendages anchored cells to the HA surface and fused with granular material covering the HA crystals. The more characteristic cellular morphology was revealed by the EDTA-KOH method. Microscopic pictures clearly identified the three-dimensional images of ruffled borders of osteoclasts and the slender cytoplasmic processes of osteocytes. This study provided further evidence for the favorable biological response of HA to bone cells as well as the value of the EDTA-KOH method in examining the stereomorphology of bone cells. PMID- 9212394 TI - Correlation between substratum roughness and wettability, cell adhesion, and cell migration. AB - Cell adhesion and spreading of chick embryo vascular and corneal explants grown on rough and smooth poly (methyl methacrylate) (PMMA) were analyzed to test the cell response specificity to substratum surface properties. Different degrees of roughness were obtained by sand-blasting PMMA with alumina grains. Hydrophilic and hydrophobic components of the surface free energy (SFE) were calculated according to Good-van Oss's model. Contact angles were determined using a computerized angle meter. The apolar component of the SFE gamma s(LW), increased with a slight roughness whereas the basic component, gamma s-, decreased. The acido-basic properties disappeared as roughness increased. Incubation of PMMA in culture medium, performed to test the influence if the biological environment, allowed surface adsorption of medium proteins which annihilated roughness effect and restored hydrophilic properties. An organotypic culture assay was carried out in an attempt to relate the biocompatibility to substratum surface state. Cell migration was calculated from the area of cell layer. Cellular adhesion was determined by measuring the kinetic of release of enzymatically dissociated cells. A slight roughness raised the migration are to an upper extent no matter which cell type. Enhancement of the cell adhesion potential was related to the degree of roughness and the hydrophobicity. PMID- 9212395 TI - The influence of alkali and alkaline earths on the working range for bioactive glasses. AB - Viscosity-temperature dependence has been investigated for glasses in a system where bioactive compositions are found. A glass is called bioactive when living bone can bond to it. In this work, high-temperature microscopy was used to determine viscosity-temperature behaviour for 40 glasses in the system Na2O-K2O MgO-CaO-B2O3-P2O5-SiO2. The silica content in the glasses was 39-70 wt% % All glasses containing < 54 mol % SiO2 devitrified during the viscosity measurements. Generally, glasses that devitrified contained more alkali but less alkaline earths than glasses with a large working range. A working range is the temperature interval at which forming of a glass can take place. This temperature interval can, for bioactive glasses, be enlarged by decreasing the amount of alkali, especially Na2O, in the glass and by increasing the amount of alkaline earths, especially MgO. Optionally, B2O3 and P2O5 can be added to the glass. An enlarged working range is a prerequisite for an expanded medical use of bioactive glasses as e.g., sintered and blown products, and fibers. PMID- 9212396 TI - Radiochemical studies on contact lens soilation. I. Lens uptake of 14C-lysozyme from simple and complex artificial tear solutions. AB - A systematic study has been made of the uptake of lysozyme by various contact lens materials. Lens uptake of 14C-methylated lysozyme was assessed using simple to complex artificial tear solutions. The data reflect prominent uptake by Group IV (ionic, high-water-content) hydrogel lenses, consistent with the literature. This includes protein on the lens surface and in the lens matrix, the former being estimated at about 33% of the total deposit for the DuraSoft 3 lenses used. Uptake appears to be contingent upon the nature of the lens material, the composition of the deposit model used, and the duration of lens exposure to the artificial tear solution. PMID- 9212397 TI - Crystal dissolution-controlled release systems. II. Metronidazole release from semicrystalline poly(vinyl alcohol) systems. AB - Novel semicrystalline, poly(vinyl alcohol) (PVA)-based phase erosion systems were developed. The rate of drug release from these systems is controlled by the rate of crystal dissolution. PVA devices loaded with metronidazole were exposed to temperatures ranging from 90 to 120 C for times of 10-90 min to obtain samples with different degrees of crystallinity. Degrees of crystallinity were measured by differential scanning calorimetry and attenuated total reflectance Fourier transform infrared spectroscopy. In vitro release of metronidazole from such systems into deionized water at 37 degrees C was monitored. The influence of parameters such as polymer molecular weight, annealing time and temperature, and surface pretreatment on the crystal dissolution, and hence the drug release rate, were investigated. Measurements of water-front movements were carried out to study the effect of parameters such as drug solubility on the release rate. The drug release rate was found to be dependent on the crystallization conditions of the samples. Surface pretreatment was found to reduce the burst effect observed during the release. PMID- 9212398 TI - Early bone formation around calcium-ion-implanted titanium inserted into rat tibia. AB - Rat tibia tissue into which calcium ion (Ca2+)-implanted titanium was surgically placed was histologically analyzed to investigate the performance of the Ca(2+) implanted titanium as a biomaterial. Calcium ions were implanted into only one side of titanium plates at 10(17) ions/cm2 and the Ca(2+)-treated titanium was surgically implanted into rat tibia for 2, 8, and 18 days. Tetracycline and calcein were used as hard-tissue labels. After excision of the tibia, the tissues were fixed, stained, embedded in polymethyl methacrylate, and sliced. The specimens were observed using a fluorescence microscope. A larger amount of new bone was formed on the Ca(2+)-treated side than on the untreated side, even at 2 days after surgery. In addition, part of the bone made contact with the Ca2(+) treated surface. On the other hand, bone formation on the untreated side was delayed and the bone did not make contact with the surface. Mature bone with bone marrow formed in 8 days. Neither macrophage nor inflammatory cell infiltration was observed. The results indicated that Ca(2+)-implanted titanium is superior to titanium alone for bone conduction. PMID- 9212399 TI - Surface display of the cholera toxin B subunit on Staphylococcus xylosus and Staphylococcus carnosus. AB - The heterologous surface expression of the cholera toxin B subunit (CTB) from Vibro cholerae in two staphylococcal species, Staphylococcus xylosus and Staphylococcus carnosus, has been investigated. The gene encoding native CTB (103 amino acids) was introduced into gene constructs encoding chimeric receptors designed to be translocated and anchored on the outer cell surface of the staphylococci. Since functionality of CTB is correlated with its ability to form pentamers and the capacity of the pentameric CTB to bind the GM1 ganglioside, both the surface accessibility and the functionality of the surface-displayed CTB receptors were evaluated. It could be concluded that the chimeric receptors were targeted to the cell wall of the staphylococci, since they could be released by lysostaphin treatment and, after subsequent affinity purification, identified as full-length products by immunoblotting. Surface accessibility of the chimeric receptors was demonstrated by a colorimetric assay and by immunofluorescence staining with a CTB-reactive rabbit antiserum. Pentamerization was investigated by using a monoclonal antibody described to be specific for pentameric CTB, and the functionality of the receptors was tested in a binding assay with digoxigenin labelled GM1. It was concluded that functional CTB was present on both types of staphylococci, and for S. carnosus, the reactivity to the pentamer-specific monoclonal antibody and in the GM1 binding assay was indeed significant. The implications of the results for the design of live bacterial vaccine delivery systems intended for administration by the mucosal route are discussed. PMID- 9212400 TI - Development of a new seminested PCR method for detection of Legionella species and its application to surveillance of legionellae in hospital cooling tower water. AB - The presence of PCR inhibitors in water samples is well known and contributes to the fact that a practical PCR assay has not been developed for legionella surveillance. In this study, we devised a new seminested PCR assay for detection of Legionella spp. in water samples as a means of overriding the PCR inhibitors without loss of sensitivity. The seminested PCR assay utilized primers to amplify the 16S rRNA gene (LEG primers) of 39 Legionella spp. The assay was specific to legionellae, and the sensitivity was 1 fg of extracted Legionella DNA in laboratory examination. To evaluate the feasibility and sensitivity of the PCR assay in identifying the presence of legionellae, it was used to survey Legionella contamination in the water of 49 cooling towers of 32 hospitals. A commercially available EnviroAmp Legionella kit and a culture method were also used in the survey for comparison with the seminested PCR assay. The detection rates of legionellae in the samples were 91.8% (45 of 49) by the PCR assay and 79.5% (39 of 49) by the culture method. The EnviroAmp kit revealed that 30.6% of the water samples (15 of 49) contained inhibitors of the PCR amplification. However, the seminested PCR assay could produce the Legionella-specific DNA bands in 14 of the 15 samples. Although 8 of the 14 samples were positive in the first step PCR, 6 of the 14 samples became positive in the second-step PCR. These results suggest that the effect of PCR inhibitors in samples, if any, can be reduced because of the dilution of the sample in the second-step PCR and that sensitivity of detection can be increased by the second-step PCR. Thus, the seminested PCR assay with LEG primers to amplify the 16S rRNA gene of 39 Legionella spp. was a practical and sensitive method to detect Legionella spp. in water samples. PMID- 9212401 TI - Cloning and characterization of two rhamnogalacturonan hydrolase genes from Aspergillus niger. AB - A rhamnogalacturonan hydrolase gene of Aspergillus aculeatus was used as a probe for the cloning of two rhamnogalacturonan hydrolase genes of Aspergillus niger. The corresponding proteins, rhamnogalacturonan hydrolases A and B, are 78 and 72% identical, respectively, with the A. aculeatus enzyme. In A. niger cultures which were shifted from growth on sucrose to growth on apple pectin as a carbon source, the expression of the rhamnogalacturonan hydrolase A gene (rhgA) was transiently induced after 3 h of growth on apple pectin. The rhamnogalacturonan hydrolase B gene was not induced by apple pectin, but the rhgB gene was derepressed after 18 h of growth on either apple pectin or sucrose. Gene fusions of the A. niger rhgA and rhgB coding regions with the strong and inducible Aspergillus awamori exlA promoter were used to obtain high-producing A. awamori transformants which were then used for the purification of the two A. niger rhamnogalacturonan hydrolases. High-performance anion-exchange chromatography of oligomeric degradation products showed that optimal degradation of an isolated highly branched pectin fraction by A. niger rhamnogalacturonan hydrolases A and B occurred at pH 3.6 and 4.1, respectively. The specific activities of rhamnogalacturonan hydrolases A and B were then 0.9 and 0.4 U/mg, respectively, which is significantly lower than the specific activity of A. aculeatus rhamnogalacturonan hydrolase (2.5 U/mg at an optimal pH of 4.5). Compared to the A enzymes, the A. niger B enzyme appears to have a different substrate specificity, since additional oligomers are formed. PMID- 9212402 TI - Ingredient selection for plastic composite supports for L-(+)-lactic acid biofilm fermentation by Lactobacillus casei subsp. rhamnosus. AB - Plastic composite supports containing 50% agricultural products (oat hulls, soybean hulls, yeast extract, soybean flour, dried bovine erythrocytes, bovine albumin, and/or mineral salts) and 50% (wt/wt) polypropylene were produced by high-temperature twin-screw extrusion. The research employed two half sets of a five-factorial fractional design (2(5 - 1)) to evaluate the effects of different agricultural components on the properties of the plastic composite supports and to select the best plastic composite support formulation for lactic acid fermentation. The biofilm population was affected by the contact angle and relative hydrophobicity of the supports (r = 0.79 to 0.82). Lactic acid was produced by the suspended cells (r = 0.96) and the biofilm on the plastic composite support discs (r = 0.85). Incorporation of yeast extract into plastic composite supports enhanced growth of free and attached cells in minimal medium (P < 0.0001). The presence of soybean hulls, yeast extract, or mineral salts in plastic composite supports produced less hydrophobic supports (P < 0.0001) and enhanced cell attachment (P < 0.03). Under all conditions, suspended-cell and polypropylene disc controls gave negligible lactic acid production and cell density. Plastic composite supports containing soybean hulls, yeast extract, soybean flour, bovine albumin, and mineral salts gave the highest biofilm population (2.3 x 10(9) CFU/g of support), cell density (absorbance of 1.8 at 620 nm), and lactic acid concentration (7.6 g/liter) in minimal medium. PMID- 9212403 TI - Optimization of L-(+)-lactic acid production by ring and disc plastic composite supports through repeated-batch biofilm fermentation. AB - Four customized bioreactors, three with plastic composite supports (PCS) and one with suspended cells (control), were operated as repeated-batch fermentors for 66 days at pH 5 and 37 degrees C. The working volume of each customized reactor was 600 ml, and each reactor's medium was changed every 2 to 5 days for 17 batches. The performance of PCS bioreactors in long-term biofilm repeated-batch fermentation was compared with that of suspended-cell bioreactors in this research. PCS could stimulate biofilm formation, supply nutrients to attached and free suspended cells, and reduce medium channelling for lactic acid production. Compared with conventional repeated-batch fermentation, PCS bioreactors shortened the lag time by threefold (control, 11 h; PCS, 3.5 h) and sixfold (control, 9 h; PCS, 1.5 h) at yeast extract concentrations of 0.4 and 0.8% (wt/vol), respectively. They also increased the lactic acid productivity of Lactobacillus casei subsp. rhamnosus (ATCC 11443) by 40 to 70% and shortened the total fermentation time by 28 to 61% at all yeast extract concentrations. The fastest productivity of the PCS bioreactors (4.26 g/liter/h) was at a starting glucose concentration of 10% (wt/vol), whereas that of the control (2.78 g/liter/h) was at 8% (wt/vol). PCS biofilm lactic acid fermentation can drastically improve the fermentation rate with reduced complex-nutrient addition. PMID- 9212404 TI - Stabilization of lignin peroxidases in white rot fungi by tryptophan. AB - Supplementation of various cultures of white rot fungi with tryptophan was found to have a large stimulatory effect on lignin peroxidase activity levels. This enhancement was greater than that observed in the presence of the lignin peroxidase recycling agent veratryl alcohol. Using reverse transcription-PCR, we found that tryptophan does not act to induce lignin peroxidase expression at the level of gene transcription. Instead, the activity enhancement observed is likely to result from the protective effect of tryptophan against H2O2 inactivation. In experiments using a partially purified lignin peroxidase preparation, tryptophan and its derivative indole were determined to function in the same way as veratryl alcohol in converting compound II, an oxidized form of lignin peroxidase, to ferric enzyme, thereby completing the catalytic cycle. Furthermore, tryptophan was found to be a better substrate for lignin peroxidase than veratryl alcohol. Inclusion of either tryptophan or indole enhanced the oxidation of the azo dyes methyl orange and Eriochrome blue black. Stimulation of azo dye oxidations by veratryl alcohol has previously been shown to be due to its enzyme recycling function. Our data allow us to propose that tryptophan stabilizes lignin peroxidase by acting as a reductant for the enzyme. PMID- 9212405 TI - Use of commercial enzyme immunoassays and immunomagnetic separation systems for detecting Escherichia coli O157 in bovine fecal samples. AB - A commercial enzyme immunoassay (EIA) (E. coli O157 Visual Immunoassay; Tecra Diagnostics) performed on enrichment cultures in modified Escherichia coli broth (mECn) was compared with immunomagnetic separation (IMS) (Dynabeads anti-E. coli O157; Dynal) performed on enrichment cultures in modified buffered peptone water (BPW-VCC) for the detection of E. coli O157 in bovine fecal samples. Tests on fecal suspensions inoculated with each of 12 different strains of E. coli O157 showed that both the EIA and IMS methods were 10- to 100-fold more sensitive than direct culture or enrichment subculture methods for detection of the organism. EIA and IMS were then compared for detection of E. coli O157 in bovine rectal swabs. For confirmation of positive EIA tests, a commercial system (Immunocapture System [ICS]; Tecra Diagnostics) was compared with IMS; both were performed on mECn enrichment cultures. Of 200 rectal swabs examined, 17 gave positive results in the EIA which were confirmed by both confirmation systems, 2 gave positive results in the EIA which were confirmed by IMS but not by ICS, and 1 gave a positive result in the EIA which was confirmed by ICS but not by IMS. Of these 20, 15 were also positive by the BPW-VCC-IMS culture system; a further 3 samples were positive by this culture system but gave a negative result in the EIA. Eight samples were negative by the BPW-VCC-IMS culture system but gave a positive result in the EIA which could not be confirmed by either confirmation system. Further examination of the eight unconfirmed EIA-positive samples yielded sorbitol-fermenting E. coli O157 from three samples. Of the remaining five cultures, four were positive in an EIA for verocytotoxins (VT) and two were positive in a cell culture assay for VT1. The remaining 170 samples were negative by both EIA and BPW-VCC-IMS. The Tecra EIA and IMS are both technically simple and sensitive methods for detecting E. coli O157 in bovine fecal samples. There was no statistically significant difference between the numbers of positives detected by the different assays (P = 0.29). PMID- 9212406 TI - Quantification of ergosterol and 3-hydroxy fatty acids in settled house dust by gas chromatography-mass spectrometry: comparison with fungal culture and determination of endotoxin by a Limulus amebocyte lysate assay. AB - Ergosterol and 3-hydroxy fatty acids, chemical markers for fungal biomass and the endotoxin of gram-negative bacteria, respectively, may be useful in studies of health effects of organic dusts, including domestic house dust. This paper reports a method for the combined determination of ergosterol and 3-hydroxy fatty acids in a single dust sample and a comparison of these chemical biomarkers determined by gas chromatography-mass spectrometry with results from fungal culture and Limulus assay. Analyses of replicate house dust samples resulted in correlations of 0.91 (ergosterol in six replicates; P < 0.01) and 0.94 (3-hydroxy fatty acids in nine replicates; P < 0.001). The amounts of ergosterol (range, 2 to 16.5 ng/mg of dust) correlated with those of total culturable fungi (range, 6 to 1,400 CFU/mg of dust) in 17 samples, (r = 0.65; P < 0.005). The amounts of endotoxin (range, 11 to 243 endotoxin units/mg of dust) measured with a modified chromogenic Limulus assay correlated with those of lipopolysaccharide (LPS) determined from 3-hydroxy fatty acid analysis of 15 samples. The correlation coefficient depended on the chain lengths of 3-hydroxy acids used to compute the LPS content. The correlation was high (r = 0.88 +/- 0.01; P < 0.001) when fatty acid chains of 10 to 14 carbon atoms were included; the correlation was much lower when hydroxy acids of 16- or 18-carbon chains were included. In conclusion, the results of the described extraction and analysis procedure for ergosterol and 3-hydroxy fatty acids are reproducible, and the results can be correlated with fungal culture and endotoxin activity of organic dust samples. PMID- 9212407 TI - Detection of DNA damage by use of Escherichia coli carrying recA'::lux, uvrA'::lux, or alkA'::lux reporter plasmids. AB - Plasmids were constructed in which DNA damage-inducible promoters recA, uvrA, and alkA from Escherichia coli were fused to the Vibrio fischeri luxCDABE operon. Introduction of these plasmids into E. coli allowed the detection of a dose dependent response to DNA-damaging agents, such as mitomycin and UV irradiation. Bioluminescence was measured in real time over extended periods. The fusion of the recA promoter to luxCDABE showed the most dramatic and sensitive responses. lexA dependence of the bioluminescent SOS response was demonstrated, confirming that this biosensor's reports were transmitted by the expected regulatory circuitry. Comparisons were made between luxCDABE and lacZ fusions to each promoter. It is suggested that the lux biosensors may have use in monitoring chemical, physical, and genotoxic agents as well as in further characterizing the mechanisms of DNA repair. PMID- 9212408 TI - Anaerobic and aerobic degradation of pyridine by a newly isolated denitrifying bacterium. AB - New denitrifying bacteria that could degrade pyridine under both aerobic and anaerobic conditions were isolated from industrial wastewater. The successful enrichment and isolation of these strains required selenite as a trace element. These isolates appeared to be closely related to Azoarcus species according to the results of 16S rRNA sequence analysis. An isolated strain, pF6, metabolized pyridine through the same pathway under both aerobic and anaerobic conditions. Since pyridine induced NAD-linked glutarate-dialdehyde dehydrogenase and isocitratase activities, it is likely that the mechanism of pyridine degradation in strain pF6 involves N-C-2 ring cleavage. Strain pF6 could degrade pyridine in the presence of nitrate, nitrite, and nitrous oxide as electron acceptors. In a batch culture with 6 mM nitrate, degradation of pyridine and denitrification were not sensitively affected by the redox potential, which gradually decreased from 150 to -200 mV. In a batch culture with the nitrate concentration higher than 6 mM, nitrite transiently accumulated during denitrification significantly inhibited cell growth and pyridine degradation. Growth yield on pyridine decreased slightly under denitrifying conditions from that under aerobic conditions. Furthermore, when the pyridine concentration used was above 12 mM, the specific growth rate under denitrifying conditions was higher than that under aerobic conditions. Considering these characteristics, a newly isolated denitrifying bacterium, strain pF6, has advantages over strictly aerobic bacteria in field applications. PMID- 9212409 TI - Strain characterization and classification of oxyphotobacteria in clone cultures on the basis of 16S rRNA sequences from the variable regions V6, V7, and V8. AB - A major problem in development of a polyphasic taxonomy is that the identification of oxyphotobacterial strains (cyanobacteria and prochlorophytes) in culture collections may be incorrect. We have therefore developed a diagnostic system using the DNA sequence polymorphism in the 16S rRNA regions V6 to V8 for individual strain characterization and identification. PCR primers amplifying V6 to V8 from oxyphotobacteria in unialgal cultures were constructed. Direct solid phase or cyclic sequencing was used to determine the sequences from the amplified DNA. This survey includes 10 strains of Nostoc/Anabaena/Aphanizomenon (Nostoc category), 5 strains of Microcystis (Microcystis category), and 4 strains of Planktothrix (Planktothrix category). Fifteen additional strains of cyanobacteria and two strains of prochlorophytes were included such that the major phyletic groups were represented. One of the strains, Phormidium sp. NIVA-CYA 203, contained an 11-nucleotide insertion with no homology to other known 16S rRNA sequences. Based on parsimony and neighbor-joining trees, the phyletic relationships of the strains were investigated. Thirteen major branches were found, with Pseudanabaena limnetica NIVA-CYA 276/6 as the most divergent strain. The strain categories Nostoc, Planktothrix, and Microcystis were all monophyletic. The sequence polymorphism within Nostoc was higher than that in Planktothrix and Microcystis. Based on the sequence and phyletic information, group-specific PCR primers for the categories Nostoc, Planktothrix, and Microcystis were constructed. For the strains included in this work, the amplifications were specific for the relevant groups. By combination of magnetic solid-phase DNA isolation and group-specific PCR amplifications, an accurate method for characterization, classification and identification of oxyphotobacterial clone cultures has been developed. PMID- 9212410 TI - Association of multiple-antibiotic-resistance profiles with point and nonpoint sources of Escherichia coli in Apalachicola Bay. AB - A total of 765 Escherichia coli isolates from point and nonpoint sources were collected from the Apalachicola National Estuarine Research Reserve, and their multiple-antibiotic-resistance (MAR) profiles were determined with 10 antibiotics. E. coli isolates from point sources showed significantly greater resistance (P < 0.05) to antibiotics and higher MAR indices than isolates from nonpoint sources. Specifically, 65 different resistance patterns were observed among point source isolates, compared to 32 among nonpoint source isolates. Examples of this contrast in MAR profiles included percentages of isolates with resistance to chlortetracycline-sulfathiazole of 33.7% and to chlortetracycline penicillin G-sulfathiazole of 14.5% for point source isolates versus 15.4 and 1.7%, respectively, for nonpoint source isolates. MAR profile homology, based on coefficient similarity, showed that isolates from point sources were markedly more diverse than isolates from nonpoint sources. Seven clusters were observed among point source isolates, with a coefficient value of approximately 1.8. In contrast, only four clusters were observed among nonpoint source isolates. Covariance matrices of data displayed six very distinct foci representing nonpoint source E. coli isolates. Importantly, E. coli isolates obtained directly from human and animal feces also clustered among point and nonpoint sources, respectively. We conclude that E. coli MAR profiles were associated with point and nonpoint sources of pollution within Apalachicola Bay and that this method may be useful in facilitating management of other estuaries. PMID- 9212411 TI - Cultivation of Escherichia coli with mixtures of 3-phenylpropionic acid and glucose: steady-state growth kinetics. AB - The fate of pollutants in the environment is affected by the presence of easily degradable carbon sources. As a step towards understanding these complex interactions, a model system was explored: the degradation of mixtures of glucose (i.e., an easily degradable substrate) and 3-phenylpropionic acid (3ppa) (a model pollutant) by Escherichia coli ML 30 was studied systematically in carbon-limited continuous culture. The two substrates were always consumed simultaneously regardless of the dilution rate applied. Even at dilution rates higher than the maximum specific growth rate for 3ppa (0.35 +/- 0.05 h-1), the two carbon substrates were utilized together. When cells were grown at a constant dilution rate with different mixtures of 3ppa and glucose, in which 3ppa contributed between 5 and 90% of carbon substrate in the feed medium, the steady-state concentrations of 3ppa and glucose were approximately proportional to the ratio of the two substrates in the feed medium. When cells were cultivated at different dilution rates with a 1:1 mixture (based on carbon) of glucose and 3ppa, an overall maximum specific growth rate of 0.90 +/- 0.05 h-1 and a Monod substrate saturation constant for 3ppa (Ks) of 600 to 700 micrograms liter-1, similar to that measured during growth with 3ppa alone, fitted the experimentally determined steady-state 3ppa concentrations. However, due to the highly differing substrate affinity constants for 3ppa and glucose (Ks approximately 30 to 70 micrograms liter-1), the total steady-state carbon concentration in the culture at a constant dilution rate was determined mainly by the steady-state 3ppa carbon concentration, and it increased with increasing proportions of 3ppa in the feed medium. PMID- 9212412 TI - Nested PCR for ultrasensitive detection of the potato ring rot bacterium, Clavibacter michiganensis subsp. sepedonicus. AB - Oligonucleotide primers derived from sequences of the 16S rRNA gene (CMR16F1, CMR16R1, CMR16F2, and CMR16R2) and insertion element IS1121 of Clavibacter michiganensis subsp. sepedonicus (CMSIF1, CMSIR1, CMSIF2, and CMISR2) were used in nested PCR to detect the potato ring rot bacterium C. michiganensis subsp. sepedonicus. Nested PCR with primer pair CMSIF1-CMSIR1 followed by primer pair CMSIF2-CMSIR2 specifically detected C. michiganensis subsp. sepedonicus, while nested PCR with CMR16F1-CMR16R1 followed by CMR16F2-CMR16R2 detected C. michiganensis subsp. sepedonicus and the other C. michiganensis subspecies. In the latter case, C. michiganensis subsp. sepedonicus can be differentiated from the other subspecies by restriction fragment length polymorphism (RFLP) analyses of the nested PCR products (16S rDNA sequences). The nested PCR assays developed in this work allow ultrasensitive detection of very low titers of C. michiganensis subsp. sepedonicus which may be present in symptomiess potato plants or tubers and which cannot be readily detected by direct PCR (single PCR amplification). RFLP analysis of PCR products provides for an unambiguous confirmation of the identify of C. michiganensis subsp. sepedonicus. PMID- 9212413 TI - Phylogeny and classification of bacteria in the genera Clavibacter and Rathayibacter on the basis of 16s rRNA gene sequence analyses. AB - A phylogenetic analysis by parsimony of 16S rRNA gene sequences (16S rDNA) revealed that species and subspecies of Clavibacter and Rathayibacter form a discrete monophyletic clade, paraphyletic to Corynebacterium species. Within the Clavibacter-Rathayibacter clade, four major phylogenetic groups (subclades) with a total of 10 distinct taxa were recognized: (I) species C. michiganensis; (II) species C. xyli; (III) species R. iranicus and R. tritici; and (IV) species R. rathayi. The first three groups form a monophyletic cluster, paraphyletic to R. rathayi. On the basis of the phylogeny inferred, reclassification of members of Clavibacter-Rathayibacter group is proposed. A system for classification of taxa in Clavibacter and Rathayibacter was developed based on restriction fragment length polymorphism (RFLP) analysis of the PCR-amplified 16S rDNA sequences. The groups delineated on the basis of RFLP patterns of 16S rDNA coincided well with the subclades delineated on the basis of phylogeny. In contrast to previous classification systems, which are based primarily on phenotypic properties and are laborious, the RFLP analyses allow for rapid differentiation among species and subspecies in the two genera. PMID- 9212414 TI - Identification of a laccase gene family in the new lignin-degrading basidiomycete CECT 20197. AB - A new lignin-degrading basidiomycete, strain I-62 (CECT 20197), isolated from decayed wood exhibited both a high dephenolization activity and decolorization capacity when tested on effluents from the sugar cane by-product fermentation industry. It has been classified as a member of the Polyporaceae family. The major ligninolytic activity detected in culture supernatants of basidiomycete I 62 was a phenoloxidase (laccase), in conjunction with small amounts of manganese peroxidase. No lignin peroxidase was detected. Laccase activity was produced in either defined or complete media. Addition of veratryl alcohol as the inducer, in defined medium, enhanced laccase production 10-fold. The use of fructose instead of glucose as a carbon source resulted in a 100-fold increase in laccase specific activity. Native isoelectrofocusing gels stained with guaiacol revealed the presence of at least seven laccase isozymes, with the most intense band being detected at pI 3. Southern hybridization analysis indicated the presence of a laccase gene family in strain I-62. Three different genes coding for phenoloxidases, lcc1, lcc2, and lcc3, were cloned and characterized. The high degree of homology between laccases from strain I-62 and laccases from Trametes species suggests a phylogenetic proximity between this new isolated fungus and the genus Trametes. PMID- 9212415 TI - Molecular microbial diversity in soils from eastern Amazonia: evidence for unusual microorganisms and microbial population shifts associated with deforestation. AB - Although the Amazon Basin is well known for its diversity of flora and fauna, this report represents the first description of the microbial diversity in Amazonian soils involving a culture-independent approach. Among the 100 sequences of genes coding for small-subunit rRNA obtained by PCR amplification with universal small-subunit rRNA primers, 98 were bacterial and 2 were archaeal. No duplicate sequences were found, and none of the sequences had been previously described. Eighteen percent of the bacterial sequences could not be classified in any known bacterial kingdom. Two sequences may represent a unique branch between the vast majority of bacteria and the deeply branching, predominantly thermophilic bacteria. Five sequences formed a clade that may represent a novel group within the class Proteobacteria. In addition, rRNA intergenic spacer analysis was used to show significant microbial population differences between a mature forest soil and an adjacent pasture soil. PMID- 9212416 TI - Production of succinic acid through overexpression of NAD(+)-dependent malic enzyme in an Escherichia coli mutant. AB - NAD(+)-dependent malic enzyme was cloned from the Escherichia coli genome by PCR based on the published partial sequence of the gene. The enzyme was overexpressed and purified to near homogeneity in two chromatographic steps and was analyzed kinetically in the forward and reverse directions. The Km values determined in the presence of saturating cofactor and manganese ion were 0.26 mM for malate (physiological direction) and 16 mM for pyruvate (reverse direction). When malic enzyme was induced under appropriate culture conditions in a strain of E. coli that was unable to ferment glucose and accumulated pyruvate, fermentative metabolism of glucose was restored. Succinic acid was the major fermentation product formed. When this fermentation was performed in the presence of hydrogen, the yield of succinic acid increased. The constructed pathway represents an alternative metabolic route for the fermentative production of dicarboxylic acids from renewable feedstocks. PMID- 9212417 TI - Strain-specific differentiation of lactococci in mixed starter culture populations using randomly amplified polymorphic DNA-derived probes. AB - A hydrophobic grid membrane filtration (HGMF) colony hybridization assay was developed that allows strain-specific differentiation of defined bacterial populations. The randomly amplified polymorphic DNA (RAPD) fingerprinting technique was used to identify potential signature nucleic acid sequences unique to each member of a commercial cheese starter culture blend. The blend consisted of two closely related Lactococcus lactis subsp. cremoris strains, 160 and 331, and one L. lactis subsp. lactis strain, 210. Three RAPD primers (OPX 1, OPX 12, and OPX 15) generated a total of 32 products from these isolates, 20 of which were potential strain-specific markers. Southern hybridization analyses revealed, that the RAPD-generated signature sequences OPX15-0.95 and a 0.36-kb HaeIII fragment of OPX1-1.0b were specific for strains 331 and 210, respectively, within the context of the test starter culture blend. These strain-specific probes were used in a HGMF colony hybridization assay. Colony lysis, hybridization, and nonradioactive detection parameters were optimized to allow specific differentiation and quantitation of the target strains in the mixed starter culture population. When the 210 and 331 probes were tested at their optimal hybridization temperatures against single cultures, they detected 100% of the target strain CFUs, without cross-reactivity to the other strains. The probes for strains 210 and 331 also successfully detected their targets in blended cultures even with a high background of the other two strains. PMID- 9212418 TI - Identification and characterization of a previously undescribed cyt gene in Bacillus thuringiensis subsp. israelensis. AB - Mosquitocidal Bacillus thuringiensis strains show as a common feature the presence of toxic proteins with cytolytic and hemolytic activities, Cyt1Aa1 being the characteristic cytolytic toxin of Bacillus thuringiensis subsp. israelensis. We have detected the presence of another cyt gene in this subspecies, highly homologous to cyt2An1, coding for the 29-kDa cytolytic toxin from B. thuringiensis subsp. kyushuensis. This gene, designated cyt2Ba1, maps upstream of cry4B coding for the 130-kDa crystal toxin, on the 72-MDa plasmid of strain 4Q2 72. Sequence analysis revealed, as a remarkable feature, a 5' mRNA stabilizing region similar to those described for some cry genes. PCR amplification and Southern analysis confirmed the presence of this gene in other mosquitocidal subspecies. Interestingly, anticoleopteran B. thuringiensis subsp. tenebrionis belonging to the morrisoni serovar also showed this gene. On the other hand, negative results were obtained with the anti-lepidopteran strains B. thuringiensis subsp. kurstaki HD-1 and subsp. aizawai HD-137. Western analysis failed to reveal Cyt2A-related polypeptides in B. thuringiensis subsp. israelensis 4Q2-72. However, B. thuringiensis subsp. israelensis 1884 and B. thuringiensis subsp. tenebrionis did show cross-reactive products, although in very small amounts. PMID- 9212419 TI - Autolysis of Lactococcus lactis caused by induced overproduction of its major autolysin, AcmA. AB - The optical density of a culture of lactococcus lactis MG1363 was reduced more than 60% during prolonged stationary phase. Reduction in optical density (autolysis) was almost absent in a culture of an isogenic mutant containing a deletion in the major autolysin gene, acmA. An acmA mutant carrying multiple coples of a plasmid encoding AcmA lysed to a greater extent than the wild-type strain did. Intercellular action of AcmA was shown by mixing end-exponential phase cultures of an acmA deletion mutant and a tripeptidase (pepT) deletion mutant. PepT, produced by the acmA mutant, was detected in the supernatant of the mixed culture, but no PepT was present in the culture supernatant of the acmA mutant. A plasmid was constructed in which acmA, lacking its own promoter, was placed downstream of the inducible promoter/operator region of the temperate lactococcal bacteriophage r1t. After mitomycin induction of an exponential-phase culture of L. lactis LL302 carrying this plasmid, the cells became subject to autolysis, resulting in the release of intracellular proteins. PMID- 9212420 TI - Identification of a novel cytochrome P-450 gene from the white rot fungus Phanerochaete chrysosporium. AB - A gene fragment belonging to the cytochrome P-450 superfamily has been cloned and identified from stationary cultures of the filamentous fungus Phanerochaete chrysosporium by reverse transcriptase (RT)-PCR. A set of degenerate primers homologous to highly conserved regions of known cytochrome P-450 sequences were used for initial RT-PCRs. Individual PCR products were cloned, sequenced, and identified as those belonging to the cytochrome P-450 superfamily based on amino acid sequence homologies and the presence of the highly conserved heme binding region. The nucleotide sequence of a single cDNA clone indicated the presence of an open reading frame encoding a partial cytochrome P-450 protein of 208 amino acids. Comparisons of the deduced amino acid sequence of the partial protein to other known cytochrome P-450 sequences indicate that it is the first member of a new family of cytochrome P-450s, designated CYP63-1A. Northern blot analysis suggests that CYP63-1A is expressed under both nitrogen-rich and nitrogen deficient culture conditions and thus not under the same regulatory constraints as the well-studied lignin and manganese peroxidases. Western blot analyses using antibodies raised to the heme binding region of CYP63-1A indicate that the protein has a molecular mass of approximately 44,000 Da. PMID- 9212421 TI - The human Lactobacillus acidophilus strain LA1 secretes a nonbacteriocin antibacterial substance(s) active in vitro and in vivo. AB - The adhering human Lactobacillus acidophilus strain LA1 inhibits the cell association and cell invasion of enteropathogens in cultured human intestinal Caco-2 cells (M. F. Bernet, D. Brassard, J. R. Neeser, and A. L. Servin, Gut 35:483-489, 1994). Here, we demonstrate that strain LA1 developed its antibacterial activity in conventional or germ-free mouse models orally infected by Salmonella typhimurium. We present evidence that the spent culture supernatant of strain LA1 (LA1-SCS) contained antibacterial components active against S. typhimurium infecting the cultured human intestinal Caco-2 cells. The LA1-SCS antibacterial activity was observed in vitro against a wide range of gram negative and gram-positive pathogens, such as Staphylococcus aureus, Listeria monocytogenes, S. typhimurium, Shigella flexneri, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Enterobacter cloacae. By contrast, no activity was observed against species of the normal gut flora, such as lactobacilli and bifidobacteria. The LA1-SCS antibacterial activity was insensitive to proteases and independent of lactic acid production. PMID- 9212422 TI - In situ analysis of nucleic acids in cold-induced nonculturable Vibrio vulnificus. AB - Low-temperature-induced nonculturable cells of the human pathogenic bacterium Vibrio vulnificus retained significant amounts of nucleic acids for more than 5 months. Upon permeabilization of fixed cells, however, an increasing number of cold-incubated cells released the nucleic acids. This indicates substantial degradation of DNA and RNA in nonculturable cells prior to fixation. Treatment of permeabilized cells with DNase and RNase allowed differential staining of DNA and RNA with the nucleic acid dye 4',6-diamidino-2-phenylindole (DAPI). Epifluorescence microscopy revealed that the could-induced nonculturable populations of V. vulnificus are highly heterogeneous with regard to their nucleic acid content. The fraction of nonculturable cells which maintained DNA and RNA structures decreased gradually during cold incubation. After 5 months at 5 degrees C, less than 0.05% of the cells could be observed to retain DNA and RNA. In parallel with the loss of nucleic acids, an increase in the concentrations of UV-absorbing material in the culture supernatants was observed in nonculturable-cell suspensions. It is hypothesized that there are two phases of the formation of nonculturable cells of V. vulnificus: the first involves a loss of culturability with maintenance of cellular integrity and intact RNA and DNA (and thus possibly viability), and the second is typified by a gradual degradation of nucleic acids, the products of which partly remain inside the cells and partly diffuse into the extracellular space. A small number of nonculturable cells, however, retain DNA and RNA, and thus may be viable despite having reduced culturability. PMID- 9212423 TI - Benzoate degradation via the ortho pathway in Alcaligenes eutrophus is perturbed by succinate. AB - During batch growth of Alcaligenes eutrophus on benzoate-plus-succinate mixtures, substrates were simultaneously metabolized, leading to a higher specific growth rate (mu = 0.56 h-1) than when a single substrate was used (mu = 0.51 h-1 for benzoate alone and 0.44 h-1 for succinate alone), without adversely affecting the growth yield (0.57 Cmol/Cmol). Flux distribution analysis revealed that succinate dehydrogenase most probably controls the rate of total succinate consumption (the maximum flux being 9.7 mmol.g-1.h-1). It is postulated that the relative consumption rate of each substrate is in part related to modified levels of gene expression but to a large extent is dependent upon the presence of succinate, end product of the beta-ketoadipate pathway. Indeed, the in vitro beta-ketoadipate succinyl coenzyme A transferase activity was seen to be inhibited by succinate, a coproduct of the reaction. PMID- 9212424 TI - Analysis of the syrP gene, which regulates syringomycin synthesis by Pseudomonas syringae pv. syringae. AB - Syringomycin is a lipodepsinonapeptide phytotoxin synthesized by Pseudomonas syringae pv. syringae on multienzymatic peptide synthetases. Sequence analysis of the interval between the syrB and syrD genes of P. syringae pv. syringae strain B301D revealed a 1,059-bp open reading frame (ORF), designated syrP. The predicted product of this ORF was a 39.6-kDa protein consisting of 353 amino acid residues. Searches of protein sequence databases demonstrated that SyrP was most similar to histidine kinases such as the CheA regulatory protein of Escherichia coli. The predicted SyrP sequence was aligned with the N terminus of CheA, a region corresponding to the phosphotransfer and acceptor domains of CheA. The SyrP region that aligns with the phosphotransfer domain of CheA contained a His at position 101 which is flanked by a weak consensus sequence of the unorthodox sensory kinase subfamily of two-component regulatory systems. Strain B301D-31, obtained by site-directed insertional mutagenesis of the syrP gene, exhibited an unusual pleiotropic phenotype including a failure to produce syringomycin in liquid media in contrast to production of elevated levels of the toxin on agar media. The syrP mutant was relieved of the suppression of toxin production that accompanies inorganic phosphate concentrations of > 1 mM on agar media. Nevertheless, the syrP mutant was substantially less virulent than the wild-type strain in pathogenicity assays in cherry fruits. These results suggest that the syrP gene encodes a regulatory protein that participates in a phosphorylation cascade controlling syringomycin production and virulence in P. syringae pv. syringae. PMID- 9212426 TI - Unique regulation of crystal protein production in Bacillus thuringiensis subsp. yunnanensis is mediated by the cry protein-encoding 103-megadalton plasmid. AB - In sporulating cultures of Bacillus thuringiensis subsp. yunnanensis HD977, two cell types are observed: cells forming only spores and cells forming only crystals. Curing analysis suggested that the crystal proteins are plasmid encoded. Through plasmid transfer experiments, it was established that a 103-MDa plasmid is involved in the crystal production. Conjugal transfer of this plasmid to Cry- recipient cells of Bacillus thuringiensis subsp. kurstaki HD73-26 conferred the ability to produce crystals exclusively on asporogenous cells of the recipient, indicating that the 103-MDa plasmid mediates the unique regulation of Cry protein production. When the dipteran-specific cryIVB gene was introduced into wild-type (Cry+) and Cry- backgrounds of B. thuringiensis subsp. yunnanensis by phage CP51ts45-mediated transduction, similar to all other B. thuringiensis strains, irregular crystals of CryIVB protein were produced by spore-forming cells in both backgrounds. However, the synthesis of the bipyramidal inclusions of B. thuringiensis subsp. yunnanensis was still limited only to asporogenous cells of the transductant. Thus, it appears that the unique property of exclusive crystal formation in asporogenous cells of B. thuringiensis subsp. yunnanensis is associated with the crystal protein gene(s) per se or its cis acting elements. As the crystals in B. thuringiensis subsp. yunnanensis were formed only in asporogenous cells, attempts were made to find out whether crystal formation had any inhibitory effect on sporulation. It was observed that both Cry+ and Cry- strains of B. thuringiensis subsp. yunnanensis (HD977 and HD977-1, respectively) exhibited comparable sporulation efficiencies. In addition, the Cry- B. thuringiensis subsp. kurstaki host (HD73-26) and its Cry+ transconjugant (HD73-26 16), expressing the B. thuringiensis subsp. yunnanensis crystal protein, were also comparable in their sporulation efficiencies, indicating that production of the crystal proteins of B. thuringiensis subsp. yunnanensis does not affect the process of sporulation. PMID- 9212425 TI - Glucose metabolism in the yeast Schwanniomyces castellii: role of phosphorylation site I and an alternative respiratory pathway. AB - Glucose metabolism in a Crabtree-negative yeast, Schwanniomyces castellii, and a cytochrome b-deficient mutant of this strain was investigated in chemostat culture. The wild-type and mutant strains exhibited the same behavior. Oxidative metabolism was observed when the substrate uptake rate (qS) was low. Fermentative metabolites were excreted when the qS value was higher than 0.40 g.g-1.h-1, indicating the occurrence of a respirofermentative metabolism; however, the respiratory quotient (RQ) remained near 1. When fermentation occurred, the cytochrome pathway was repressed but not the salicylhydroxamic acid (SHAM) sensitive pathway. The presence of an alternative SHAM-sensitive respiratory pathway and the presence of phosphorylation site I in all metabolic conditions explained the RQ value of 1 and accounted for high biomass yields in oxidative metabolism conditions (0.62 g.g-1 for the wild-type strain and 0.31 g.g-1 for the cytochrome b-deficient mutant strain). PMID- 9212427 TI - Biochemical and molecular characterization of the insecticidal fragment of CryV. AB - Two C-terminal deletion constructs were made to study the effect of such deletions on the biological activity of the CryV protein of Bacillus thuringiensis subsp. kurstaki. The results of feeding on neonatal larvae of Ostrinia nubilalis (European corn borer [ECB]) indicated that the 50% lethal dose of the full-length CryV protein was 3.34 micrograms/g of diet (95% fiducial limits, 2.53 to 4.32 micrograms/g of diet). Removal of 71 amino acids (aa) from the C terminus had little effect on toxicity, whereas deletion of 184 aa abolished the insecticidal activity of the CryV protein completely. Truncations of the full-length CryV protein were also generated with trypsin and the midgut protease of ECB. The proteolytically treated products were characterized by determining their N-terminal amino acid sequences. The CryV protein was found to be cleaved by both proteases through a two-step process. Initially an intermediary form was generated which contained aa 45 of full-length CryV as its N-terminal end. The C-terminal end of this peptide was not experimentally determined. However, analysis of the deduced amino acid sequence of CryV indicated that the C-terminal end of the intermediary form is likely either aa 655 or 659. Further N-terminal processing of the intermediary form resulted in a protease-resistant core form. The core included aa 156 to aa 655 or 659. While the intermediary form retained 100% of the ECB larval toxicity, the core form exhibited only approximately 22% of the toxicity of the full-length protein. PMID- 9212428 TI - Molecular microbial diversity of an anaerobic digestor as determined by small subunit rDNA sequence analysis. AB - The bacterial community structure of a fluidized-bed reactor fed by vinasses (wine distillation waste) was analyzed. After PCR amplification, four small subunit (SSU) rDNA clone libraries of Bacteria, Archaea, Procarya, and Eucarya populations were established. The community structure was determined by operational taxonomic unit (OTU) phylogenetic analyses of 579 partial rDNA sequences (about 500 bp long). A total of 146 OTUs were found, comprising 133, 6, and 7 from the Bacteria, Archaea, and Eucarya domains, respectively. A total of 117 bacterial OTU were affiliated with major phyla: low-G+C gram-positive bacteria, Cytophaga-Flexibacter-Bacteroides, Proteobacteria, high-G+C gram positive bacteria, and Spirochaetes, where the clone distribution was 34, 26, 17, 6, and 4%, respectively. The other 16 bacterial OTUs represent 13% of the clones. They were either affiliated with narrow phyla such as Planctomyces-Chlamydia, green nonsulfur bacteria, or Synergistes, or deeply branched on the phylogenetic tree. A large number of bacterial OTUs are not closely related to any other hitherto determined sequences. The most frequent bacterial OTUs represents less than 5% of the total bacterial SSU rDNA sequences. However, the 20 more frequent bacterial OTUs describe at least 50% of these sequences. Three of the six Archaea OTUs correspond to 95% of the Archaea population and are very similar to already known methanogenic species: Methanosarcina barkeri, Methanosarcina frisius, and Methanobacterium formicicum. In contrast, the three other Archaea OTUs are unusual and are related to thermophilic microorganisms such as Crenarchaea or Thermoplasma spp. Five percent of the sequences analyzed were chimeras and were removed from the analysis. PMID- 9212429 TI - Use of the pre-pro part of Staphylococcus hyicus lipase as a carrier for secretion of Escherichia coli outer membrane protein A (OmpA) prevents proteolytic degradation of OmpA by cell-associated protease(s) in two different gram-positive bacteria. AB - Heterologous protein secretion was studied in the gram-positive bacteria Bacillus subtilis and Staphylococcus carnosus by using the Escherichia coli outer membrane protein OmpA as a model protein. The OmpA protein was found to be translocated across the plasma membrane of both microorganisms. However, the majority of the translocated OmpA was similarly degraded in B. subtilis and S. carnosus despite the fact that the latter organism does not secrete soluble exoproteases into the culture medium. The finding that purified OmpA, which was added externally to the culture medium of growing S. carnosus cells, remained intact indicates that newly synthesized and exported OmpA is degraded by one or more cell-associated proteases rather than by a soluble exoprotease. Fusion of the mature part of OmpA to the pre-pro part of a lipase from Staphylococcus hyicus allowed the efficient release of the corresponding propeptide-OmpA hybrid protein into the supernatant and completely prevented the cell-associated proteolytic degradation of the mature OmpA, most likely reflecting an important function of the propeptide during secretion of its natural mature lipase moiety. The relevance of our findings for the biotechnological use of gram-positive bacteria as host organisms for the secretory production of heterologous proteins is discussed. PMID- 9212430 TI - Nitrogen availability of grape juice limits killer yeast growth and fermentation activity during mixed-culture fermentation with sensitive commercial yeast strains. AB - The competition between selected or commercial killer strains of type K2 and sensitive commercial strains of Saccharomyces cerevisiae was studied under various conditions in sterile grape juice fermentations. The focus of this study was the effect of yeast inoculation levels and the role of assimilable nitrogen nutrition on killer activity. A study of the consumption of free amino nitrogen (FAN) by pure and mixed cultures of killer and sensitive cells showed no differences between the profiles of nitrogen assimilation in all cases, and FAN was practically depleted in the first 2 days of fermentation. The effect of the addition of assimilable nitrogen and the size of inoculum was examined in mixed killer and sensitive strain competitions. Stuck and sluggish wine fermentations were observed to depend on nitrogen availability when the ratio of killer to sensitive cells was low (1:10 to 1:100). A relationship between the initial assimilable nitrogen content of must and the proportion of killer cells during fermentation was shown. An indirect relationship was found between inoculum size and the percentage of killer cells: a smaller inoculum resulted in a higher proportion of killer cells in grape juice fermentations. In all cases, wines obtained with pure-culture fermentations were preferred to mixed-culture fermentations by sensory analysis. The reasons why killer cells do not finish fermentation under competitive conditions with sensitive cells are discussed. PMID- 9212431 TI - A spontaneous change in the intracellular cyclic AMP level in Aspergillus niger is influenced by the sucrose concentration in the medium and by light. AB - A spontaneous rise in intracellular cyclic AMP (cAMP) levels was observed in the early stages of Aspergillus niger growth under conditions yielding large amounts of citric acid. The amount of cAMP formed was found to depend on the initial concentration of sucrose in the medium. Under higher-sucrose conditions, the cAMP peak appeared earlier and was higher, while in lower-sucrose media a flattened peak was observed later in fermentation. Since in media with higher concentrations of sucrose intracellular citric acid starts to accumulate earlier and more rapidly, cAMP synthesis may be triggered by intracellular acidification, which is caused by the dissociation of citric acid. No spontaneous increase in cAMP concentrations could be detected when the cells were grown in continuously illuminated cultures, suggesting that A. niger phosphodiesterase (PDE) is photoregulated. More evidence for the activation of PDE by light was obtained from morphological studies under light and dark conditions in the presence of cAMP or N6,O2'-dibutyryl cAMP, and this idea was additionally supported by experiments in which PDE inhibitors were tested. PMID- 9212432 TI - Molecular genetic characterization of the L-lactate dehydrogenase gene (ldhL) of Lactobacillus helveticus and biochemical characterization of the enzyme. AB - The Lactobacillus helveticus L-(+)-lactate dehydrogenase (L-LDH) gene (ldhL) was isolated from a lambda library. The nucleotide sequence of the ldhL gene was determined and shown to have the capacity to encode a protein of 323 amino acids (35.3 kDa). The deduced sequence of the 35-kDa protein revealed a relatively high degree of identity with other lactobacillar L-LDHs. The highest identity (80.2%) was observed with the Lactobacillus casei L-LDH. The sizes and 5' end analyses of ldhL transcripts showed that the ldhL gene is a monocistronic transcriptional unit. The expression of ldhL, studied as a function of growth, revealed a high expression level at the logarithmic phase of growth. The ldhL gene is preceded by two putative -10 regions, but no corresponding -35 regions could be identified. By primer extension analysis, the ldhL transcripts were confirmed to be derived from the -10 region closest to the initiation codon. However, upstream of these regions additional putative -10/-35 regions could be found. The L-LDH was overexpressed in Escherichia coli and purified to homogeneity by two chromatographic steps. The purified L-LDH was shown to be a nonaliosteric enzyme, and amino acid residues involved in allosteric regulation were not conserved in L. helveticus L-LDH. However, a slight enhancement of enzyme activity was observed in the presence of fructose 1,6-diphosphate, particularly at neutral pH. A detailed enzymatic characterization of L-LDH was performed. The optimal reaction velocity was at pH 5.0, where the kinetic parameters K(m), and Kcat for pyruvate were 0.25 mM and 643 S-1, respectively. PMID- 9212433 TI - Thiram and dimethyldithiocarbamic acid interconversion in Saccharomyces cerevisiae: a possible metabolic pathway under the control of the glutathione redox cycle. AB - A rapid decrease of intracellular glutathione (GSH) was observed when exponentially growing cells of Saccharomyces cerevisiae were treated with sublethal concentrations of either dimethyldithiocarbamic acid or thiram [bis(dimethylthiocarbamoyl) disulfide]. The underlying mechanism of this effect possibly involves the intracellular oxidation of dimethyldithiocarbamate anions to thiram, which in turn oxidizes GSH. Overall, a linear relationship was found between thiram concentrations up to 21 microM and production of oxidized GSH (GSSG). Cytochrome c can serve as the final electron acceptor for dimethyldithiocarbamate reoxidation, and it was demonstrated in vitro that NADPH handles the final electron transfer from GSSG to the fungicide by glutathione reductase. These cycling reactions induce transient alterations in the intracellular redox state of several electron carriers and interfere with the respiration of the yeast. Thiram and dimethyldithiocarbamic acid also inactivate yeast glutathione reductase when the fungicide is present within the cells as the disulfide. Hence, whenever the GSH regeneration rate falls below its oxidation rate, the GSH:GSSG molar ratio drops from 45 to 1. Inhibition of glutathione reductase may be responsible for the saturation kinetics observed in rates of thiram elimination and uptake by the yeast. The data suggest also a leading role for the GSH redox cycle in the control of thiram and dimethyldithiocarbamic acid fungitoxicity. Possible pathways for the handling of thiram and dimethyldithiocarbamic acid by yeast are considered with respect to the physiological status, the GSH content, and the activity of glutathione reductase of the cells. PMID- 9212434 TI - A novel means to develop strain-specific DNA probes for detecting bacteria in the environment. AB - A simple means to develop strain-specific DNA probes for use in monitoring the movement and survival of bacteria in natural and laboratory ecosystems was developed. The method employed amplification of genomic DNA via repetitive sequence-based PCR (rep-PCR) using primers specific for repetitive extragenic palindromic (REP) elements, followed by cloning of the amplified fragments. The cloned fragments were screened to identify those which were strain specific, and these were used as probes for total genomic DNA isolated from microbial communities and subjected to rep-PCR. To evaluate the utility of the approach, we developed probes specific for Burkholderia cepacia G4 and used them to determine the persistence of the strain in aquifer sediment microcosms following bioaugmentation. Two of four probes tested were found to specifically hybridize to DNA fragments of the expected sizes in the rep-PCR fingerprint of B. cepacia G4 but not to 64 genetically distinct bacteria previously isolated from the aquifer. One of these probes, a 650-bp fragment, produced a hybridization signal when as few as 10 CFU of B. cepacia G4 were present in a mixture with 10(6) CFU nontarget strains, indicating that the sensitivity of these probes was comparable to those of other PCR-based detection methods. The probes were used to discriminate groundwater and microcosm samples that contained B. cepacia G4 from those which did not. False-positive results were obtained with a few samples, but these were readily identified by using hybridization to the second probe as a confirmation step. The general applicability of the method was demonstrated by constructing probes specific to three other environmental isolates. PMID- 9212435 TI - Phylogenetic analysis and in situ identification of bacteria in activated sludge. AB - The bacterial community structure of activated sludge of a large municipal wastewater treatment plant was investigated by use of the rRNA approach. Almost full-length genes coding for the small-subunit rRNA (rDNA) were amplified by PCR and subsequently cloned into the pGEM-T vector. Clones were screened by dot blot hybridization with group-specific oligonucleotide probes. The phylogenetic affiliations of clones were compared with the results obtained with the original sample by in situ hybridization with fluorescently labeled, rRNA-targeted oligonucleotide probes and found to be in general agreement. Twenty-five 16S rDNA clones were fully sequenced, 11 were almost fully (> 80%) sequenced, and 27 were partially sequenced. By comparative sequence analyses, the majority of the examined clones (35 of 67) could be affiliated with the beta subclass of the class Proteobacteria. The gamma and alpha subclasses of Proteobacteria were represented by 13 and 4 clones, respectively. Eight clones grouped with the epsilon group of Proteobacteria, and five clones grouped with gram-positive bacteria with a low DNA G+C content. The 16S rDNA of two clones showed similarity with 16S rDNA genes of members of the phyla Chlamydiae and Planctomyces. 16S rRNA targeted oligonucleotide probes were designed and used for the enumeration of the respective bacteria. Interestingly, potentially pathogenic representatives of the genus Arcobacter were present in significant numbers (4%) in the activated sludge sample examined. Pairs of probes targeted to the 5' and 3' regions were used for detection of chimeric sequences by in situ hybridization. Two clones could be identified as chimera by applying such a pair of probes. PMID- 9212436 TI - Development of the FUN-1 family of fluorescent probes for vacuole labeling and viability testing of yeasts. AB - A new family of fluorescent probes has been developed for assessing the viability and metabolic activity of yeasts. This class of halogenated unsymmetric cyanine dyes is exemplified by the FUN-1 [2-chloro-4-(2,3-dihydro-3-methyl-(benzo-1,3 thiazol-2-yl)- methylidene)-1-phenylquinolinium iodide] stain, a membrane permeant nucleic acid-binding dye that has been found to give rise to cylindrical intravacuolar structures (CIVS) in Saccharomyces cerevisiae. Biochemical processing of the dye by active yeasts yielded CIVS that were markedly red shifted in fluorescence emission and therefore spectrally distinct from the nucleic acid-bound form of the dye. The formation of CIVS occurred under both aerobic and anaerobic conditions and was highly temperature dependent. Treatment of yeasts with the nonmetabolizable glucose analog 2-deoxy-D-glucose reduced cellular ATP levels approximately 6-fold and completely inhibited CIVS formation. Under aerobic conditions, the formation of CIVS was abrogated by the cytochrome oxidase inhibitors azide and cyanide; however, the H+ transport uncoupler carbonyl cyanide m-chlorophenylhydrazone inhibited CIVS formation under both aerobic and anaerobic conditions. Depletion of cellular thiols, including glutathione, with millimolar concentrations of N-ethylmaleimide, iodoacetamide, or allyl alcohol completely inhibited CIVS production. Marked reduction in the formation of CIVS by ethacrynic acid and sulfobromophthalein, inhibitors of glutathione S-transferase, suggested that dye processing can involve enzyme mediated formation of glutathione conjugates. The conversion of FUN-1 by S. cerevisiae was studied quantitatively by using several techniques, including fluorometry, flow cytometry, and wide-field and confocal laser scanning fluorescence microscopy. PMID- 9212437 TI - Novel metabolites in phenanthrene and pyrene transformation by Aspergillus niger. AB - Aspergillus niger, isolated from hydrocarbon-contaminated soil, was examined for its potential to degrade phenanthrene and pyrene. Two novel metabolites, 1 methoxyphenanthrene and 1-methoxypyrene, were identified by conventional chemical techniques. Minor metabolites identified were 1- and 2-phenanthrol and 1-pyrenol. No 14CO2 evolution was observed in either [14C]phenanthrene or [14C]pyrene cultures. PMID- 9212438 TI - Conservation of plasmid-encoded dibenzothiophene desulfurization genes in several rhodococci. AB - The cloned sulfur oxidation (desulfurization) genes (sox) for dibenzothiophene (DBT) from the prototype Rhodococcus sp. strain IGTS8 were used in Southern hybridization and PCR experiments to establish the DNA relatedness in six new rhodococcal isolates which are capable of utilizing DBT as a sole sulfur source for growth. The ability of these strains to desulfurize appears to be an exclusive property of a 4-kb gene locus on a large plasmid of ca. 150 kb in IGTS8 and ca. 100 kb in the other strains. Besides a difference in plasmid profile, IGTS8 is distinguishable from the other strains in at least the copy number of the insertion sequence IS1166, which is associated with the sox genes. PMID- 9212439 TI - Growth of silicone-immobilized bacteria on polycarbonate membrane filters, a technique to study microcolony formation under anaerobic conditions. AB - A technique was developed to study microcolony formation by silicone-immobilized bacteria on polycarbonate membrane filters under anaerobic conditions. A sudden shift to anaerobiosis was obtained by submerging the filters in medium which was depleted for oxygen by a pure culture of bacteria. The technique was used to demonstrate that preinduction of nitrate reductase under low-oxygen conditions was necessary for nonfermenting, nitrate-respiring bacteria, e.g., Pseudomonas spp., to cope with a sudden lack of oxygen. In contrast, nitrate-respiring, fermenting bacteria, e.g., Bacillus and Escherichia spp., formed microcolonies under anaerobic conditions with or without the presence of nitrate and irrespective of aerobic or anaerobic preculture conditions. PMID- 9212440 TI - Expression and functional analysis of a hyperglycosylated glucoamylase in a parental host, Aspergillus awamori var. kawachi. AB - A modified glucoamylase gene (glaA) with an extra Thr- and Ser-rich Gp-I domain (T. Semimaru, M. Goto, K. Furukawa, and S. Hayashida, Appl. Environ. Microbiol. 61:2885-2890, 1995) was introduced into a mutant parental host, Aspergillus awamori var. kawachi, in which the original glaA gene had been completely deleted and replaced with the hygromycin phosphotransferase gene. The modified glaA was successfully expressed and secreted. The modified glucoamylase possessed higher digestibility of raw corn starch and higher stabilities in response to heat and extreme pH. PMID- 9212441 TI - PCR-mediated detection of acidophilic, bioleaching-associated bacteria. AB - The detection of acidophilic microorganisms from mining environments by culture methods is time consuming and unreliable. Several PCR approaches were developed to amplify small-subunit rRNA sequences from the DNA of six bacterial phylotypes associated with acidic mining environments, permitting the detection of the target DNA at concentrations as low as 10 fg. PMID- 9212442 TI - Clostridium thermocellum JW20 (ATCC 31549) is a coculture with Thermoanaerobacter ethanolicus. AB - A PCR assay based on 16S rRNA sequence differences among four thermophilic anaerobic bacterial strains was used to demonstrate contamination of Clostridium thermocellum JW20 (ATCC 31549) with a Thermoanaerobacter ethanolicus strain. Therefore, we suggest that interpretation of experimental results with C. thermocellum JW20 be viewed with caution. PMID- 9212443 TI - Bacterial diversity in Adirondack mountain lakes as revealed by 16S rRNA gene sequences. AB - Bacterial communities of seven lakes in the Adirondack Mountains of New York State were characterized by amplification and sequencing of 16S ribosomal DNA. Analysis of over 100 partial sequences revealed a diverse collection of lineages, largely of the class Proteobacteria (19% alpha subdivision, 31% beta subdivision, and 9% gamma subdivision), the phylum Cytophaga-Flavobacteria-Bacteroides (15%), and the order Actinomycetales (18%). Additionally, a number of the sequences were similar to those of the order Verrucomicrobiales. However, few of the sequence types are closely related to those of characterized species. The relative contributions of the groups of sequences differed among the lakes, suggesting that bacterial population structure varies and that it may be possible to relate aquatic bacterial community structure to water chemistry. PMID- 9212445 TI - Post-hatching development of circular mantle muscles in the squid Loligo opalescens. AB - Post-hatching development of the circular muscles in the mantle of squid was studied morphometrically to identify structural changes and to quantify hyperplasia and hypertrophy of the muscle fibers. Superficial, mitochondria-rich (SMR) fibers and central, mitochondria-poor (CMP) fibers are present at hatching. Although both fiber types increase in size and, even more so, in number during post-hatching development, CMP fibers increase at a much higher rate than do SMR fibers. As a result, the relative proportion of SMR to CMP fibers shifts from about 1:1 in a hatchling to about 1:6 in an 8-week-old animal; it then apparently remains constant to adulthood. These structural changes are consistent with developmental changes in muscular activity. During slow, jet-propelled swimming, 1-week-old animals show mantle contractions that have twice the relative amplitude and frequency of those in adults. The presence of Na-channel protein in mantle muscle was detected bio-chemically by using site-directed antibodies; the protein was found to be preferentially expressed in CMP fibers. These results suggest that SMR fibers are an important source of locomotory power at hatching, but become progressively less important during the first 8 weeks of development as CMP fibers assume the dominant role in jet locomotion. PMID- 9212446 TI - Fine structure of the apical ganglion and its serotonergic cells in the larva of Aplysia californica. AB - The apical ganglion is a highly conserved structure present in various marine invertebrate larvae. Although one of the hallmarks of this ganglion is the presence of serotonergic cells, little is known about the structure and function of these cells. We have examined this ganglion in larvae of the marine mollusc Aplysia with light- and electron-microscopic immunocytochemistry. The results indicate that the cellular composition of the apical ganglion of Aplysia is very similar to that of other opisthobranchs. It consists of three classes of sensory cells (ampullary, para-ampullary, and ciliary tuft cells) and of other nerve cell types. Almost a third of the cells in the apical ganglion of Aplysia are serotonergic, and these can be divided into two classes: three para-ampullary and two interneuronal cells. All of the serotonergic cells extend an axon into the central nervous system. The variety of sensory and serotonergic cell types suggests that each type processes distinct attributes of the sensory environment. We argue that the apical ganglion, by virtue of its serotonergic cells, is well suited to play important roles in the integration of sensory information to achieve proper motor adaptation to variable seawater conditions. PMID- 9212444 TI - Embryogenesis in hydra. AB - Embryogenesis in hydra includes a variable period of dormancy; and this period, as well as subsequent stages through hatching, takes place within a thick cuticle that hinders observation. Thus, although the early stages of development have been well-characterized qualitatively, the middle and later stages are only poorly understood. Here, we provide a detailed description of the stages of embryogenesis, including the time required to traverse each of the stages, and the changes that occur in the type and number of cells throughout the stages. The events of cleavage and gastrulation occur within the first 48 h. Cleavage is holoblastic and unipolar and leads to a single-layered coeloblastula. Gastrulation occurs by ingression and is followed by the deposition of the thick cuticle. Thereafter, during the variable period of dormancy ranging from 2-24 weeks, little occurs; the important events are the conversion of the outer layer into an ectoderm and the appearance of the interstitial cell lineage. During the last 2 days before hatching, the endoderm and gastric cavity form, while stem cells of the interstitial cell lineage proliferate and differentiate into neurons, nematocytes, and secretory cells. Finally, the cuticle cracks, and the hatchling enlarges and emerges from the cuticle as a functional animal. The formation of the gastric cavity and the hatching of the embryo are both explicable in terms of the osmotic behavior of the animal and the hydrostatic forces generated by this behavior. Characteristics of development that are common to hydra and triploblastic phyla are presented. PMID- 9212447 TI - Preparation and local anaesthetic activity of N-[2-(tert-amino)ethyl]- and N (lupinyl)-benzotriazol-1/2-ylacetamides. AB - Two sets of N-[2-(tert-amino)ethyl]- and N-[(quinolizidin-1 alpha-yl) methyl] benzotriazol-2-ylacetamides, bearing substituents on position 5 or 5 and 6, were prepared and tested for local anaesthetic activity in comparison with lidocaine. Most of the prepared compounds exhibited a fairly good activity comparable or superior to that of lidocaine. The introduction of substituents on the benzene ring and the replacement of the usual tert-amino alkyl chains with the quinolizidin-1 alpha-ylmethyl (lupinyl) moiety were quite profitable for both the intensity and duration of activity. One selected compound was subjected to a large pharmacological screening and found endowed with a good level of the purported antiarrhythmic activity without any other disturbing activity. PMID- 9212448 TI - New benzamide-derived 5-HT3 receptor antagonists which prevent the effects of ethanol on extracellular dopamine, and fail to reduce voluntary alcohol intake in rats. AB - A set of substituted benzamides, characterized by the presence of a bulky quinolizidine moiety, were subjected to binding assays for 5-HT3 and D2 receptors on membranes obtained from the bovine area postrema ([3H]-GR65630) and the rat striatum ([3H]-spiperone) respectively. These benzamides resulted unsuitable for the recognition of D2 receptors, while a few of them, devoid of 5-HT4 receptor activity, had consistent affinity for central 5-HT3 receptors, inhibiting also potently the ethanol-induced dopamine efflux from the mesolimbic dopamine terminal region. However they failed in attenuating voluntary alcohol consumption in rats, as observed with several other chemically unrelated 5-HT3 antagonists. Thus the 5-HT3-mediated inhibition of alcohol-induced striatal release of dopamine by substituted benzamides is not a requisite for affecting ethanol intake. PMID- 9212449 TI - Nonpeptide angiotensin II receptor antagonists: synthesis and biological activity of 1H-imidazo and 1H-[1,2,3]-triazolo fused derivatives. AB - A series of 1H-Imidazo and 1H-[1,2,3]-Triazolo fused derivatives have been obtained and tested as non peptide AII receptor antagonists. Modification of the classical biphenyl moiety to a 4-arylthienyl fragment afforded interesting activities. PMID- 9212450 TI - Quinoxaline chemistry. Part 7. 2-[aminobenzoates]- and 2-[aminobenzoylglutamate] quinoxalines as classical antifolate agents. Synthesis and evaluation of in vitro anticancer, anti-HIV and antifungal activity. AB - Thirty-three quinoxalines bearing an aminobenzoyl or aminobenzoylglutamate group on position 2 and various substituents on position 3,6,7 of the heterocycle were prepared in order to evaluate in vitro anticancer activity. Preliminary screening performed at NCI showed that most derivatives exhibited a moderate to strong growth inhibition activity on various tumor panel cell lines between 10(-5) and 10(-4) Molar concentrations. Interesting selectivities were also recorded between 10(-8) and 10(-6) M. Among the series examined one compound (29) which was the most active also exhibited both in vitro anti-HIV protection and antifungal activity while in other two (31, 37) the antifungal activity was prevailing. PMID- 9212451 TI - Synthesis of 4,5-functionalized-2-methyl-6-(substituted aryl)-3 (2H) pyridazinones: a new group of potent platelet aggregation inhibitors. AB - A series of 4,5-functionalized-2-methyl-6-(substituted phenyl)-3 (2H) pyridazinones were synthesized and evaluated as platelet aggregation inhibitors in human platelet rich plasma (PRP). The new products generally displayed significant higher activity with respect to the corresponding unsubstituted aryl compounds. Compounds 27 and 31 appeared of particular interest, being their IC50s in the submicromolar range. Structure-activity relationships (SARs) are discussed. PMID- 9212452 TI - Tricyclic heteroaromatic systems. 3-substituted 1,2,3,5,6,7-hexahydro-2,5,6 trioxopyrazino[1,2,3-de]-quinoxalines as novel antagonists at the glycine-NMDA site. AB - Two novel antagonists of the glycine-NMDA receptor have been synthesized and tested for their ability to displace [3H]glycine from its specific binding site in rat brain cortical membranes. The 3-substituted pyrazino[1,2,3-de]quinoxalin 2,5,6-triones 1a-b contain all the essential pharmacophoric descriptors of a glycine receptor antagonist. A model is proposed for the binding of these compounds that includes some of the interactions postulated for the potent glycine-NMDA receptor antagonist L-689,560. PMID- 9212453 TI - Synthesis and pharmacological activities of some 2,3,4,5 tetrahydro[1,5]benzo[f]thiazepines. AB - In search of new biological active agents in the series of [1,5]benzothiazepines, some 2,3,4,5-tetrahydro-N-(5-morpholinopentanoyl)-[1,5]benzo[f] thiazepines were synthesized and examined in vitro for their calcium antagonist activity compared to the Diltiazem one. PMID- 9212454 TI - The radioprotective power of 2-isopropyl 5-methyl 1,3-thiazolane. AB - The adequate conditions of radioprotective treatment with 2-isopropyl 5-methyl 1,3-thiazolane were established for mice. Protection is maximum with a unique intraperitoneal injection of LD 50/2, 3 hours prior to the irradiation. The rate of radiation dose reduction is therefore 1.75. Survival rate of whole body irradiated mice with supralethal doses were determined for 11 months. The long term survival of the animals fully proved good prevention of radio-induced damages. In vitro pharmacological studies show the ability of the major metabolite of the compound to trap organic radicals. PMID- 9212455 TI - A new indirect spectrofluorimetric determination of sub-micromolar concentrations of manganese(II) in bovine liver by means of 2-hydroxyquinoline. AB - A new high-sensitive and at the same time accurate and well documented indirect spectrofluorimetric method for the determination of sub-micromolar concentrations of manganese(II) being as a normal trace element in the bovine liver, was described. The purpose of the experimental work is performed by means of two fluorescence intensity measurements; the measurement of the fluorescence (F1) of a 2.0 x 10(-5) mol/l solution of pure 2-hydroxyquinoline (2-HQ) (lambda ex: 320 nm: lambda em: 375 nm), and that of the fluorescence of the above solution after the addition of Mn(II) to be analyzed (F2). The observed difference of the fluorescence intensities measured (F1-F2) = delta F, is exclusively owed to the quenching of the Mn(II) on the free 2-HQ. The presence of various metal ions considerably interferes with the above described determination of Mn(II). For this reason, the separation and/or masking of these undesirable metal ions it is advisable to apply before any analytical praxis. PMID- 9212456 TI - The length of the vertebral column of primates: an allometric study. AB - The length of the vertebral column of 425 primates (151 prosimians, 76 platyrrhines and 198 catarrhines) was related to body mass from bibliographic sources. Regressions were calculated for the whole sample and separately for the three taxonomic groups quoted above. In parallel, the lengths of the cervical, thoracic and lumbar regions were calculated in a sample of 105 primates (30 prosimians, 19 platyrrhines and 56 catarrhines) and partial correlations established. In all cases except one, the correlation coefficients were significant. Of these, 12 correlations (out of 16) scaled with negative allometry (< 0.33), 4 with positive allometry (> 0.33), and in 6 cases the exponents were not significantly different from the criterion for isometry (0.33). The lumbar region showed the highest variability, mainly in platyrrhines and catarrhines. Results from catarrhines are globally the closest to the expectations of elastic similarity. No obvious direct relationship was found between the length of the vertebral column and the number of vertebrae. PMID- 9212457 TI - Mothers in a wild group of moor macaques (Macaca maurus) are more attractive to other group members when holding their infants. AB - Social interactions between mothers that were holding their infants and other group members were studied in a wild group of moor macaques (Macaca maurus). Affiliative approaches to infants (AAI) by group members that were accompanied with particular behaviours, such as mouthing and touching or affiliative grunts, were observed frequently. Females approached mothers with infants more frequently than did males in all age classes. Female reproductive status, parity and dominance relationships between interactants had little effect on the frequency of approaches. Mothers received more grooming and performed less grooming when they were holding their infants than when they were not. A high level of social tolerance in macaque species with relaxed dominance styles might allow the free expression of AAI. Dominance styles among females and AAI could be linked via a positive feedback relationship since AAI might play a role in relaxing the dominance style among females. PMID- 9212458 TI - First discovery of the hairy-eared dwarf lemur (Allocebus trichotis) in a highland rain forest of eastern Madagascar. PMID- 9212459 TI - Possible involvement of sexual selection in neocortical evolution of monkeys and apes. PMID- 9212460 TI - Conspecific aggression and predation: costs for a solitary mantled howler monkey. PMID- 9212461 TI - Genes involved in organ separation in Arabidopsis: an analysis of the cup-shaped cotyledon mutant. AB - Mutations in CUC1 and CUC2 (for CUP-SHAPED COTYLEDON), which are newly identified genes of Arabidopsis, caused defects in the separation of cotyledons (embryonic organs), sepals, and stamens (floral organs) as well as in the formation of shoot apical meristems. These defects were most apparent in the double mutant. Phenotypes of the mutants suggest a common mechanism for separating adjacent organs within the same whorl in both embryos and flowers. We cloned the CUC2 gene and found that the encoded protein was homologous to the petunia NO APICAL MERISTEM (NAM) protein, which is thought to act in the development of embryos and flowers. PMID- 9212462 TI - Plant viral synergism: the potyviral genome encodes a broad-range pathogenicity enhancer that transactivates replication of heterologous viruses. AB - Synergistic viral diseases of higher plants are caused by the interaction of two independent viruses in the same host and are characterized by dramatic increases in symptoms and in accumulation of one of the coinfecting viruses. In potato virus X (PVX)/potyviral synergism, increased pathogenicity and accumulation of PVX are mediated by the expression of potyviral 5' proximal sequences encoding P1, the helper component proteinase (HC-Pro), and a fraction of P3. Here, we report that the same potyviral sequence (termed P1/HC-Pro) enhances the pathogenicity and accumulation of two other heterologous viruses: cucumber mosaic virus and tobacco mosaic virus. In the case of PVX-potyviral synergism, we show that the expression of the HC-Pro gene product, but not the RNA sequence itself, is sufficient to induce the increase in PVX pathogenicity and that both P1 and P3 coding sequences are dispensable for this aspect of the synergistic interaction. In protoplasts, expression of the potyviral P1/HC-Pro region prolongs the accumulation of PVX (-) strand RNA and transactivates expression of a reporter gene from a PVX subgenomic promoter. Unlike the synergistic enhancement of PVX pathogenicity, which requires only expression of HC-Pro, the enhancement of PVX ( ) strand RNA accumulation in protoplasts is significantly greater when the entire P1/HC-Pro sequence is expressed. These results indicate that the potyviral P1/HC Pro region affects a step in disease development that is common to a broad range of virus infections and suggest a mechanism involving transactivation of viral replication. PMID- 9212463 TI - Activation of the silent psbA1 gene in the cyanobacterium Synechocystis sp strain 6803 produces a novel and functional D1 protein. AB - The photosystem II reaction center protein D1 in Synechocystis sp strain 6803 is encoded by the psbA2 and psbA3 genes of the three-membered psbA gene family. The silent and divergent psbA1 copy of the psbA gene family was activated by exchanging part of its upstream region with a corresponding fragment of the psbA2 copy. The light-regulated expression of the activated psbA1 gene showed that the inserted psbA2 segment contains the information necessary for light-dependent as well as high-light-stimulated transcription. The activated psbA1 gene expressed a novel D1 protein, D1'. A mutant strain containing psbA1 as the only active psbA gene grew photoautotrophically at a rate comparable to that of the wild type. This finding demonstrates that despite its unusual amino acid sequence, D1 is exchangeable for D1 in the photosystem II complex, at least under normal laboratory conditions. The D1' protein was found to have a degradation rate similar to that of the D1 protein under low- or high-light conditions. Another mutant containing the activated psbA1 gene together with the psbA2 and psbA3 genes produced both the D1 and D1' proteins. PMID- 9212465 TI - A role for sugar transporters during seed development: molecular characterization of a hexose and a sucrose carrier in fava bean seeds. AB - To analyze sugar transport processes during seed development of fava bean, we cloned cDNAs encoding one sucrose and one hexose transporter, designated VfSUT1 and VfSTP1, respectively. sugar uptake activity was confirmed after heterologous expression in yeast. Gene expression was studied in relation to seed development. Transcripts were detected in both vegetative and seed tissues. In the embryo, VfSUT1 and VfSTP1 mRNAs were detected only in epidermal cells, but in a different temporal and spatial pattern. VfSTP1 mRNA accumulates during the midcotyledon stage in epidermal cells covering the mitotically active parenchyma, whereas the VfSUT1 transcript was specific to outer epidermal cells showing transfer cell morphology and covering the storage parenchyma. Transfer cells developed at the contact area of the cotyledonary epidermis and the seed coat, starting first at the early cotyledon stage and subsequently spreading to the abaxial region at the late cotyledon stage. Feeding high concentrations of sugars suppressed both VfSUT1 expression and transfer cell differentiation in vitro, suggesting a control by carbohydrate availability. PMID- 9212464 TI - The Arabidopsis downy mildew resistance gene RPP5 shares similarity to the toll and interleukin-1 receptors with N and L6. AB - Plant disease resistance genes operate at the earliest steps of pathogen perception. The Arabidopsis RPP5 gene specifying resistance to the downy mildew pathogen Peronospora parasitica was positionally cloned. It encodes a protein that possesses a putative nucleotide binding site and leucine-rich repeats, and its product exhibits striking structural similarity to the plant resistance gene products N and L6. Like N and L6, the RPP5 N-terminal domain resembles the cytoplasmic domains of the Drosophila Toll and mammalian interleukin-1 transmembrane receptors. In contrast to N and L6, which produce predicted truncated products by alternative splicing, RPP5 appears to express only a single transcript corresponding to the full-length protein. However, a truncated form structurally similar to those of N and L6 is encoded by one or more other members of the RPP5 gene family that are tightly clustered on chromosome 4. The organization of repeated units within the leucine-rich repeats encoded by the wild-type RPP5 gene and an RPP5 mutant allele provides molecular evidence for the heightened capacity of this domain to evolve novel configurations and potentially new disease resistance specificities. PMID- 9212466 TI - Modification of seed oil content and acyl composition in the brassicaceae by expression of a yeast sn-2 acyltransferase gene. AB - A putative yeast sn-2 acyltransferase gene (SLC1-1), reportedly a variant acyltransferase that suppresses a genetic defect in sphingolipid long-chain base biosynthesis, has been expressed in a yeast SLC deletion strain. The SLC1-1 gene product was shown in vitro to encode an sn-2 acyltransferase capable of acylating sn-1 oleoyl-lysophosphatidic acid, using a range of acyl-CoA thioesters, including 18:1-, 22:1-, and 24:0-CoAs. The SLC1-1 gene was introduced into Arabidopsis and a high erucic acid-containing Brassica napus cv Hero under the control of a constitutive (tandem cauliflower mosaic virus 35S) promoter. The resulting transgenic plants showed substantial increases of 8 to 48% in seed oil content (expressed on the basis of seed dry weight) and increases in both overall proportions and amounts of very-long-chain fatty acids in seed triacylglycerols (TAGs). Furthermore, the proportion of very-long-chain fatty acids found at the sn-2 position of TAGs was increased, and homogenates prepared from developing seeds of transformed plants exhibited elevated lysophosphatidic acid acyltransferase (EC 2.3.1.51) activity. Thus, the yeast sn-2 acyltransferase has been shown to encode a protein that can exhibit lysophosphatidic acid acyltransferase activity and that can be used to change total fatty acid content and composition as well as to alter the stereospecific acyl distribution of fatty acids in seed TAGs. PMID- 9212467 TI - Epigenetic silencing of a foreign gene in nuclear transformants of Chlamydomonas. AB - The unstable expression of introduced genes poses a serious problem for the application of transgenic technology in plants. In transformants of the unicellular green alga Chlamydomonas reinhardtii, expression of a eubacterial aadA gene, conferring spectinomycin resistance, is transcriptionally suppressed by a reversible epigenetic mechanism(s). Variations in the size and frequency of colonies surviving on different concentrations of spectinomycin as well as the levels of transcriptional activity of the introduced transgene(s) suggest the existence of intermediate expression states in genetically identical cells. Gene silencing does not correlate with methylation of the integrated DNA and does not involve large alterations in its chromatin structure, as revealed by digestion with restriction endonucleases and DNase I. Transgene repression is enhanced by lower temperatures, similar to position effect variegation in Drosophila. By analogy to epigenetic phenomena in several eukaryotes, our results suggest a possible role for (hetero)chromatic chromosomal domains in transcriptional inactivation. PMID- 9212468 TI - Effects of synergistic signaling by phytochrome A and cryptochrome1 on circadian clock-regulated catalase expression. AB - Persistent oscillation in constant conditions is a defining characteristic of circadian rhythms. However, in plants transferred into extended dark conditions, circadian rhythms in mRNA abundance commonly damp in amplitude over two or three cycles to a steady state level of relatively constant, low mRNA abundance. In Arabidopsis, catalase CAT3 mRNA oscillations damp rapidly in extended dark conditions, but unlike catalase CAT2 and the chlorophyll a/b binding protein gene CAB, in which the circadian oscillations damp to low steady state mRNA abundance, CAT3 mRNA oscillations damp to high steady state levels of mRNA abundance. Mutational disruption of either phytochrome- or cryptochrome-mediated light perception prevents damping of the oscillations in CAT3 mRNA abundance and reveals strong circadian oscillations that persist for multiple cycles in extended dark conditions. Damping of CAT3 mRNA oscillations specifically requires phytochrome A but not phytochrome B and also requires the cryptochrome1 blue light receptor. Therefore, we conclude that synergistic signaling mediated through both phytochrome A and cryptochrome1 is required for damping of circadian CAT3 mRNA oscillations in extended dark conditions. PMID- 9212472 TI - Inhibitors of polygalacturonase in calli of Orobanche aegyptiaca. AB - The presence of at least three distinct polygalacturonases (PGase) in callus of Orobanche was demonstrated. The PGase activity is labile and at pH 4.5 does not require activation by cations. It can be partially purified on Biogel P 100 columns and can be resolved by PAGE into several bands. Broomrape callus tissue also contains inhibitors of PGase activity. One of these is a low M(r) compound, stable to boiling and removable by dialysis. An additional inhibitor precipitable by ammonium sulphate is also present. PMID- 9212469 TI - Light-stimulated degradation of an unassembled Rieske FeS protein by a thylakoid bound protease: the possible role of the FtsH protease. AB - Unassembled subunits of the cytochrome b6f complex as well as components of other unassembled chloroplastic complexes are rapidly degraded within the organelle. However, the mechanisms involved in these proteolytic processes are obscure. When the Rieske FeS protein (RISP) is imported into isolated chloroplasts in vitro, some of the protein does not property assemble with the cytochrome complex, as determined by its sensitivity to exogenous protease. When assayed in intact, lysed, or fractionated chloroplasts, the imported RISP was found to be sensitive to endogenous proteases as well. The activity responsible for degradation of the unassembled protein was localized to the thylakoid membrane and characterized as a metalloprotease requiring zinc ions for its activity. The degradation rate was stimulated by light, but no involvement of ATP or redox control was observed. Instead, when the RISP that was attached to thylakoid membranes was first illuminated on ice, degradation proceeded in either light or darkness at equal rates suggesting a light-induced conformational change making the protein prone to degradation. Antibodies raised against native FtsH, a bacterial, membrane bound, ATP-dependent, zinc-stimulated protease, effectively inhibited degradation of the unassembled RISP, suggesting a role for the chloroplastic FtsH in this process. PMID- 9212473 TI - 13-phenyltridecanoic acid in seed lipids of some aroids. AB - Several closely related long-chain omega-phenylalkanoic and omega-phenylalkenoic acids occur in the seed lipids of genera of the subfamily Aroideae of the Araceae. One, 13-phenyltridecanoic acid, is a major component. This is the first report of these acids in plant lipids. Their presence in only one subfamily may indicate that the Araceae is diphyletic. PMID- 9212470 TI - Retrotransposon insertion into the maize waxy gene results in tissue-specific RNA processing. AB - We previously reported that three alleles of the maize waxy (wx) gene were alternatively spliced as a result of the insertion of retrotransposons into intronic sequences. In addition, inefficient splicing of element sequences with the surrounding intron produced wild-type transcripts that presumably were responsible for the observed residual gene expression in the endosperm. In this study, we report that one of these alleles, wxG, has a tissue-specific phenotype with 30-fold more WX enzymatic activity in pollen than in the endosperm. Quantification of wxG-encoded transcripts in pollen and the endosperm demonstrates that this difference can be accounted for by tissue-specific differences in RNA processing. Specifically, there is approximately 30-fold more correctly spliced RNA in pollen than in the endosperm. Based on an analogy to similar examples of tissue-specific alternative splicing in animal systems, we hypothesize that the tissue-specific phenotype of the wxG allele may reflect differences in the concentration of splicing factors in these tissues. PMID- 9212474 TI - The incidence of post dural puncture headache in Taiwanese patients undergoing cesarean section. AB - BACKGROUND: In Taiwan, there was only a retrospective study about the post-dural puncture headache (PDPH) resulting from spinal anesthesia for cesarean section (C/S), but it did not mention the relationship between the incidence of PDPH and the number of dural punctures, as well as between the gauge of spinal needle. Therefore, we designed a prospective study to investigate if the spinal needles for smaller gauges could decrease the incidence of PDPH in anesthesia for C/S. METHODS: From Jan. 1990 to June 1991 we prospectively observed 2,385 consecutive cases of spinal anesthesia for various types of surgical procedures, of which 584 were C/S. The spinal needles used were of gauges 24, 25 and 26. In practice, needles of these gauges were randomly applied. The PDPH was observed until its disappearance, and nonexistence of PDPH was also followed for at least one week. All of the data were analyzed using the Fisher exact test. RESULTS: The overall incidence of PDPH was 1.18%. The incidence of PDPH in C/S females was 3.08%, which was significantly higher than that in non-obstetric females (0.37%). Although the incidence in all females (1.31%) was significantly higher than that in males (0.71%), the incidence in non-obstetric females (0.37%) did not differ significantly compared with males. The incidence of PDPH relevant to the gauges of spinal needle used was not statistically different in C/S females. CONCLUSIONS: It appears that the incidence of PDPH does not differ between Taiwanese and Westerners. Pregnancy may be the key factor contributing to higher incidence of PDPH. The 26-gauge spinal needle may lower the incidence of PDPH to a greatest extent in C/S patients, although in comparison with 24- and 25-gauge needles the difference is not statistically significant. PMID- 9212475 TI - Metabolic characteristics and enflurane defluorination of cytochrome P450 dependent monooxygenases in human hepatocellular carcinoma. AB - BACKGROUND: Xenobiotic metabolism and defluorination capacity of microsomal monooxygenases were investigated in vitro through the surgical specimens of liver resected from patients with hepatocellular carcinoma and patients of extrahepatic pathology as control. METHODS: In microsomes of hepatocellular carcinoma tissues, the activities of cytochrome P450-dependent monooxygenase isoenzymes 1A1, 2B1, and 2E1 were evaluated in vitro by reacting with the specific marker substrates benzo(a)pyrene, benzphetamine and aniline, respectively, in the generating incubation system. The distant normal liver tissues and tissues from control patients with extrahepatic lesion were also investigated for comparison. The ability of enflurane defluorination was assessed by Orion combined for detection of free fluoride ion production. RESULTS: Concentrations of P450 total content, cytochrome b5, and NADPH-cytochrome c reductase showed parallel and marked reduction in tumor tissues when compared with its distant normal regions or normal livers. The monooxygenase functions displayed significant decreases within the tumor tissues as benzo(a)pyrene hydroxylation > or = benzphetamine demethylation > aniline hydroxylation in magnitude. Defluorination of enflurane also markedly decreased in tumor tissues comparing with normal livers. CONCLUSIONS: These marked reductions in the compositions and in vitro metabolic activities, including defluorination of anesthetics, in the cytochrome P450 dependent monooxygenases within the tumor tissues characterize the unique pattern of xenobiotic metabolism in patients with hepatocellular carcinoma. PMID- 9212476 TI - Priming technique accelerates the onset time of mivacurium in children during halothane anesthesia. AB - BACKGROUND: Mivacurium is considered a relaxant suitable for tracheal intubation in children due to its rapid onset. We compared the neuromuscular effects of mivacurium, with and without priming, in children undergoing elective surgery during halothane anesthesia. METHODS: Forty pediatric patients (2-10 yr, ASA class I) were randomly into 2 groups and studied under halothane anesthesia. The non-priming group (n = 20) received mivacurium 0.25 mg/kg, and the priming group (n = 20) received a priming dose of mivacurium 0.025 mg/kg, followed by an intubating dose of 0.225 mg/kg 3 min later. Thenar Electromyogram responsive to supramaximal train-of-four stimulation of the ulnar nerve at 12 s intervals was used as neuromuscular monitoring. RESULTS: The onset time in the priming group was significantly faster than in the non-priming group (1.04 min vs. 1.7 min). The mean time from injection of intubating dose to spontaneous recovery to 25%, 50% and 75% twitch were not influenced by priming technique. Side effects, such as cutaneous flushing and hypotension, were unremarkable at this dose in children. CONCLUSIONS: Priming technique can significantly accelerates the onset of mivacurium in the pediatric patients under halothane anesthesia. PMID- 9212477 TI - Comparison of sevoflurane with halothane in pediatric ambulatory anesthesia: an experience in Taiwan. AB - BACKGROUND: Sevoflurane, with blood/gas partition coefficient of 0.69 and MAC of 1.76 is a fast acting, potent inhalation anesthetic. Its suitability and safety for pediatric ambulatory anesthesia were assessed and compared with that of halothane. METHODS: Thirty two unpremedicated pediatric patients undergoing elective herniorrhaphy on day-surgery basis were randomly allocated to 2 groups of equal number to receive either sevoflurane or halothane anesthesia. Employing mask technique, anesthesia was induced with 60% nitrous oxide and 3 MAC of either sevoflurane or halothane in oxygen. Anesthesia was maintained respectively with 1 1.5 MAC of sevoflurane or halothane. The induction time, emergence time and untoward effects during anesthesia were analyzed and compared. RESULTS: It was shown that both the induction time and emergence time were significantly shorter in patients receiving sevoflurane. None had major complications. CONCLUSIONS: The results strongly suggest that sevoflurane is preferable to halothane for pediatric ambulatory anesthesia. PMID- 9212478 TI - Comparison of oral controlled-release morphine with transdermal fentanyl in terminal cancer pain. AB - BACKGROUND: Controlled-release morphine (MST) given twice daily provides a simpler and more convenient treatment regimen than 4-hourly opioid administration for the control of cancer pain. Recently, a new formulation of transdermal fentanyl (TDF) has been developed which provides a new route for the treatment of cancer pain. The present study was designed to compare the analgesic efficacy, safety and adverse effects of MST and TDF in the management of chronic cancer pain. METHODS: In this open, comparative and randomized study, patients were treated with oral morphine hydrochloride immediate-release (MHIR) in the stabilization phase and then the prescription was switched to MST or TDF for 14 days in the treatment phase. Oral MHIR was provided as rescue medication for breakthrough pain. Assessments of the pain intensity, pain frequency, degree of pain improvement, profile of mood states, quality of sleep, activity status and adverse effects were performed before and after the stabilization phase and before and after the treatment phase. RESULTS: Forty of 47 cancer patients completed the study with 20 patients in each group. There were significant (p < 0.05) improvements in pain intensity, pain frequency, mood states and quality of sleep in both groups before and after treatment, while improvement in the activity status was not significant. No specific adverse effects were encountered except for drowsiness which occurred in 6 patients treated with MST and 5 treated with TDF (p < 0.05). Insomnia was significantly improved (p < 0.05) with both regimens compared with that in the period before treatment. There were no significant differences between the two study groups in analgesic efficacy or adverse effects. CONCLUSIONS: These results suggest that TDF and MSt are safe and effective analgesics for the management of chronic cancer pain. However, TDF provides a simpler and more convenient treatment for those patients with severe nausea, vomiting or dysphagia. PMID- 9212479 TI - Post dural puncture headache in cesarean section: comparison of 25-gauge Whitacre with 25- and 26-gauge Quincke needles. AB - BACKGROUND: Our previous study showed that there were no significant differences in the incidence of post dural puncture headache (PDPH) relevant to the use of 24 to 26-gauge Quincke spinal needles in obstetric patients. Again, we were eager to know if the pencil-point spinal needle (Whitacre) would be able to decrease the incidence of PDPH compared to Quincke spinal needle. METHODS: We prospectively observed 94 spinal anesthesias for cesarean section performed during the period from May 1993 to July 1995. The 25-gauge Whitacre needles were used. In practice the insertion of needle was made through median line approach and the puncture was considered eligible only in one attempt. The PDPH was observed until its disappearance, and one without PDPH had also been observed for at least one week for likelihood of delayed occurrence. The data were compared with those of our previous study regarding the use of 25- and 26-gauge Quincke needles in obstetric patients. All of the data were analyzed using the Fisher exact test. RESULTS: The incidence of PDPH was 1.06%. In comparison there was no significant difference from that of 25- and 26-gauge Quincke needles (3.65% and 2.06%, respective). Only one case suffered from PDPH in the Whitacre group. It was mild and relieved with bed rest and hydration. CONCLUSIONS: Although the difference was not statistically significant, the 25-gauge Whitacre spinal needle caused a lower incidence and less severity of PDPH than the 25- and 26-gauge Quincke needles did. PMID- 9212480 TI - Epidural anesthesia for emergency caesarean section in a patient with single ventricle and aortic stenosis. AB - A 24-year-old parturient with single ventricle and moderate aortic stenosis was admitted due to preeclampsia and fetal distress at 31 weeks' gestation. Emergency Caesarean section was performed under lumbar epidural anesthesia and epidural analgesia was given for post-operative pain control. Mother and baby both survived. The anesthetic techniques and managements in other parturients with similar congenital cardiac anomalies are also reviewed and described. PMID- 9212481 TI - Absorption of irrigating fluid during transcervical resection of endometrium--a report of two cases. AB - It has been recognized for many years that the use of hypotonic solution for the irrigation of the bladder cavity during transurethral resection of the prostate (TURP) may result in hyponatremia and water intoxication due to rapid and excessive absorption of the solution from the exposed prostatic bed, the clinical manifestation of which is termed "TURP syndrome". A similar condition termed "female TURP syndrome" following hysteroscopic transcervical endometrial resection (TCR) has been reported. Since the frequency of TCR continues to increase the increased rate of "TCR syndrome" would come in its wake. Here, we present two cases who developed severe hyperglycemia and hyponatremia while underwent TCR with 10% dextrose in water as the irrigation fluid and the same time emphasize the potential risk of this complication. PMID- 9212482 TI - High frequency jet ventilation for severe pulmonary hemorrhage during aortic dissection operation--a case report. AB - Massive endobronchial bleeding during extracorporeal circulation was encountered in a patient during aortic dissection operation. The use of high frequency jet ventilation resulted in successful staunching of the bleeding, thus avoiding the need of pneumonectomy or lobectomy. PMID- 9212483 TI - The lost endotracheal tube--a rare complication of accidental esophageal intubation. AB - Endotracheal intubation is a discreet skill for the management of compromised airway. Various complications associated with this procedure have been described. Here, we would like to present a pediatric patient suffering from cerebellar atrophy, who was intubated in a local clinic due to seizure with cyanosis and loss of consciousness. Unfortunately, due to inadvertent esophageal intubation and bad management, the patient swallowed the endotracheal tube together with two detached loose teeth. This irrational and iatrogenic medical misconduct as exemplified in this accident calls forth the need of educating and disciplining the nonanesthetic physicians for acute management of airway, particularly of those who would likely come across difficult airway problems. PMID- 9212484 TI - Rehabilitative, psychiatric, functional and aesthetic problems in patients treated for burn injuries--a preliminary follow-up study. AB - Patients consecutively treated for burn injuries for four or more days during 1994 were examined one year after admission by a plastic surgeon, a specialist in rehabilitation medicine and a psychiatrist. Of thirty-nine such patients treated, two were dead, 11 did not present and six thought they had no remaining problems. Aesthetic and functional problems were present in 16 patients, in 11 reconstructive surgery given in one or more sessions was judged to have improved the condition. Of eighteen patients referred to a rehabilitation medicine specialist, 14 were assessed. Nine of these had functional impairments in the burn-injured body regions. A majority had functional impairments, persistent decrease in range of upper extremity motion, reduced muscle force, altered sensibility and itch. One patient suffered from pain. Three patients had occupational handicaps. Work disability occurred in two patients and further two were in need of vocational counselling due to the burn injury. In a subgroup of 11 patients four fulfilled criteria for one or more personality disorders, and two of these also suffered from major depression. Quality of life assessed with the SF-36 was lower than in a normal population. Some of the patients had psychiatric disease and personality disorders. Although rehabilitation started early in the acute phase of treatment, rehabilitation medicine function increasing measures were needed in several cases. Individual rehabilitation programmes based on the patient's particular features and needs are recommended. The findings support the idea of a multidisciplinary approach for patients with burn injury and indicate that a subgroup of burn injury patients have functional impairments and/or disabilities which can probably be improved with reconstructive surgery and rehabilitation. PMID- 9212485 TI - Rationale for early tangential excision and grafting in burn patients. AB - Early excision and grafting of the burn wound in the first 9 days remain the keys to survival for patients with major burn injuries. In the last 7-year-period, 54 major burn cases were treated in our burn facility, the only Burn Center in Istanbul. Early excision and grafting were performed to 32 of them, admitted in the first week. Others were admitted later and managed conservatively. Their follow-up results in terms of mortality and morbidity rates were compared. In addition to improvement in the prognosis, early excision and grafting procedures decreased the duration of hospitalization and cost of burn treatment. PMID- 9212486 TI - The effects and uses of heparin in the care of burns that improves treatment and enhances the quality of life. AB - Burn care has been mostly surgical, difficult, and expensive. The use of adequately large doses of heparin administered both parenterally and topically improved burn treatment and quality of life (1-20). Burn pain was relieved. Patients were not toxic. Tissue swelling, resuscitation fluids, and healing time were reduced. Pulmonary and intestinal pathology were notably absent. The new skin was smooth, comfortable, and contracture free. More than anticoagulating effect were seen. Antiinflammatory effects, active at acidic phs but not at alkaline phs, stopped burn pain inflammation, and extension. Neoangiogenic effects revascularized ischemic tissue. Reepithelializing effects were evident. Nonburn and recent burn studies confirmed that heparin had these strong properties and effects (7-13, 21-34). Medical and surgical procedures were reduced. Burn care became simpler and easier. Precautions to prevent bleeding were used. Current use of heparin is limited. Wider use is warranted. PMID- 9212487 TI - Prevention and therapy of postburn scars. AB - The cosmetic and functional result in postburn scar deformities is influenced by following factors: 1. The type of patient's central nervous system and his response to burn injury. 2. Depth and site of burn areas. 3. Early excision and grafting. 4. Infection complications, their severity and location. 5. Fixation of dressings should be done using elastic materials and applied for so long until stabilisation of scars is completed. Elastic materials should be combined with rigid pressure and pressure massage. 6. Congenital predisposition of the patient to hypertrophic scarring. PMID- 9212488 TI - Burns and epilepsy. AB - This is a report of the first descriptive analytic study of a group of 183 burn patients, treated in the Burn Unit at the University Hospital of Cartagena, Colombia during the period since January 1985 until December 1990. There is presented experience with the selected group of 24 patients in whom the diagnosis of burn was associated with epilepsy. There is also analysed and described the gravity of the scars sequels, neurological disorders, the complication of the burn and an impact of this problem on the patient, his (her) family and the community. It is very important to report that there was found Neurocisticercosis in 66.6% of the group of burn patients with epilepsy, and it is probably the first risk factor of burn in this group. PMID- 9212489 TI - Fluid resuscitation in thermally injured pediatric patients. AB - More than two-thirds of critical burns in special burn units are children. The burned child continues to represent a special challenge, since resuscitation therapy must be more precise than that for an adult with a similar burn. Children have a limited physiologic reserve and the pediatric fluid replacement therapy is based on the principle of separate calculation of physiological and pathological losses. We have reviewed the most widely accepted pediatric isotonic fluid protocols. All these protocols calculate for replacement of pathological losses with a need of 2 ml/kg/% BSAB (body surface area burn) or 4 ml/kg/% BSAB. We choosed the formulas of two Shriner's Burns Institutes--the Cincinnati and the Galveston Unit as representatives, and calculated the fluid therapy for model burn children weights of 10 kg, 30 kg with 20, 40, 60, 80% BSAB. The results of calculations where compared with physiologic parameters of children. In conclusions we could show, that the 4 ml/kg/% BSAB formulas do replace all theoretically predicted pathophysiologic losses due to burns. However, the 2 ml/kg/% BSAB formulas are more practical as a guideline for resuscitation of pediatric patients because of greater therapeutical range and better clinical response of children threatened by burn shock. It is important to remember that all formulas are only guides to fluid therapy, they should be modified according to individual needs and clinical status of the patient. Only successful restoring and maintaining perfusion pressures leads to optimal oxygenation of injured and noninjured tissues, which promotes spontaneous healing, prevents wound conversion, minimise bacterial colonisation, and prepares the injured areas for early grafting. PMID- 9212490 TI - Abstract genetic representation of dynamical neural networks using Kauffman networks. AB - Abstract (developmental) genetic representations are schemes where each genotype encodes a program for the construction of a phenotype. A new method for contriving abstract genetic representations is presented, based upon Kauffman's ideas regarding biological development [13-16]. Phenogenesis is controlled by genotype via cell replication and differentiation. Comparison is made with the earlier published methods of Gruau [9] and Kitano [18]. It is argued that greater expressive power is obtained using Kauffman networks. The new method was tested in the artificial evolution of morphology and finally successfully applied to the synthesis of structure in dynamical neural networks. PMID- 9212491 TI - Evolutionary stagnation due to pattern-pattern interactions in a coevolutionary predator-prey model. AB - We consider a spatially structured model of a coevolutionary predator-prey system with interactions in a one-dimensional phenotype space. We show that in phenotype space predators and prey organize themselves into distinct clusters of phenotypes called quasi-species. The prey quasi-species also cluster in patches in real space. As the prey quasi-species evolve away from the predator quasi-species (in phenotype space) the prey patch size reduces and the single predator quasi species is inhibited from evolving toward either of the two prey species. We show that it is the interaction between the phenotype space patterns (quasi-species) and the real space patterns (patches) that inhibit the predators from evolving. PMID- 9212492 TI - Evolutionary transitions and artificial life. AB - A major challenge for artificial life is to synthesize the evolutionary transitions that have repeatedly formed differentiated higher-level entities from cooperative organizations of lower-level entities, producing the nested hierarchical structure of living processes. This article identifies the key elements and relationships that must be incorporated or synthesized in an artificial life system if these transitions are to emerge. The processes currently included in artificial life systems are unable to provide an adequate basis for the emergence of the complex cooperative organization that is essential to the transitions. A new theory of the evolution of cooperative organization is developed that points to the additional processes that must be included in artificial life systems to underpin the emergence of the transitions. PMID- 9212493 TI - Multiple von Neumann computers: an evolutionary approach to functional emergence. AB - A novel system composed of multiple von Neumann computers and an appropriate problem environment is proposed and simulated. Each computer has a memory to store the machine instruction program, and when a program is executed, a series of machine codes in the memory is sequentially decoded, leading to register operations in the central processing unit (CPU). By means of these operations, the computer not only can handle its generally used registers but also can read and write the environmental database. Simulation is driven by genetic algorithms (GAs) performed on the population of program memories. Mutation and crossover create program diversity in the memory, and selection facilitates the reproduction of appropriate programs. Through these evolutionary operations, advantageous combinations of machine codes are created and fixed in the population one by one, and the higher function, which enables the computer to calculate an appropriate number from the environment, finally emerges in the program memory. In the latter half of the article, the performance of GAs on this system is studied. Under different sets of parameters, the evolutionary speed, which is determined by the time until the domination of the final program, is examined and the conditions for faster evolution are clarified. At an intermediate mutation rate and at an intermediate population size, crossover helps create novel advantageous sets of machine codes and evidently accelerates optimization by GAs. PMID- 9212494 TI - Molecular mechanisms involved in T-B interactions. PMID- 9212495 TI - Tracing antigen-driven B cell development in humans. PMID- 9212496 TI - T cells in the selection of germinal center B cells. PMID- 9212497 TI - Biased VH4 gene segment repertoire in the human tonsil. PMID- 9212498 TI - Subepithelial B cells of the human tonsil. PMID- 9212499 TI - Malignant lymphomas stem from different B cell populations. PMID- 9212501 TI - B cell populations: the multiple myeloma model. PMID- 9212500 TI - What do chronic B cell malignancies teach us about B cell subsets? PMID- 9212502 TI - Regulation of human B cell growth and differentiation: lessons from the primary immunodeficiencies. PMID- 9212503 TI - Alternative therapies. PMID- 9212504 TI - Practice patterns. PMID- 9212506 TI - Knowledge, leaders and progress, 1968. PMID- 9212505 TI - Johns Hopkins address: does nursing have a future? AB - PURPOSE: To outline the importance of identifying where nursing is heading as a profession and explore the metaphors it uses to explain its role to society. Current images of nursing are confused. Nurses should articulate clearly why nursing should be retained as a social force for improving health and meeting the needs of vulnerable people. SCOPE: The historical development of nursing considering the metaphors used. CONCLUSIONS: Because of contemporary pressures on nursing, the metaphors used are inconsistent and require interpretation. Nurses should find new metaphors and resolve complex internal tensions. Unless nursing can articulate its primary function more cogently, it risks extinction. PMID- 9212507 TI - Leadership in practice, 1980. PMID- 9212508 TI - Comparison of nursing interventions classification and current procedural terminology codes for categorizing nursing activities. AB - PURPOSE: To compare the frequency with which nursing activity terms could be categorized using Nursing Interventions Classification (NIC) and Current Procedural Terminology (CPT) codes. DESIGN: Descriptive. The sample was 201 patients with AIDS hospitalized 1989-1992 for pneumocystis carinii pneumonia in three US medical centers. METHODS: Nursing activity terms (n = 21,366) were collected from patient interviews, nurse interviews, intershift reports, and patient records, then were categorized using NIC and CPT codes. RESULTS: Nursing activity terms were categorized into 80 NIC interventions across 22 classes and into 15 CPT codes. All terms in the data set were classifiable using the NIC system and the majority (60%) of the terms were classified into 14 NIC intervention categories; 6% of the terms were classifiable by CPT codes. The most frequently used CPT code was "pulse oximetry." Significantly (p < .0001) greater numbers of nursing activity terms could be categorized in the NIC system compared to the CPT system. CONCLUSIONS: Findings provide evidence that NIC is superior to CPT for categorizing nursing activities in this study's population. The findings support the importance of discipline-specific classifications for categorization of health care interventions. Nursing-specific intervention classification systems such as NIC, the Omaha System, and the Home Health Care Classification are essential to defining the contribution of nursing to both quality and cost outcomes. PMID- 9212509 TI - Balancing engagement and detachment in caregiving. AB - PURPOSE: To investigate how caregivers balance engagement with detachment to cope with cumulative demands and losses. DESIGN: Qualitative, descriptive. Population, formal and informal caregivers in the United States. A sample of 14 was studied between 1992 and 1994. METHODS: Data were collected in open-ended interviews, then were coded and analyzed using grounded-theory methods. Credibility and fittingness were established. RESULTS: Caregivers who balance engagement and detachment can affect outcomes without needing to control outcomes. Such caregivers are pragmatic and make conscious choices based on their emotional needs. They set and maintain limits and boundaries and are able to monitor the balancing process while recognizing the importance of practicing self-care. CONCLUSIONS: The longer and more intense a caregiver's involvement, the more important it is to learn to balance engagement and detachment. Caregivers may need to learn effective balancing skills. PMID- 9212510 TI - Change and continuity in family caregiving practices with young mothers and their children. AB - PURPOSE: To examine how young mothers who gave birth during adolescence extended and developed caregiving practices within the context of family relationships, caregiving traditions, and life events. DESIGN: Longitudinal, interpretive phenomenological. A community-based sample in 1993 consisted of 13 of the 16 young mothers and 11 of the 18 grandparents who had participated in a 1988 study. Three male partners of the young mothers also participated in this 1993 study. Families resided in a Western metropolitan area in the United States. METHODS: Life history accounts of the intervening years, stories of family routines, and recent coping episodes of parenting were elicited through in-depth interviews with the young mothers and their male partners; one interview was conducted with grandparents. Data were analyzed using the interpretive approach. FINDINGS: Adversarial caregiving practices develop or change in the context of transformed family relationships. CONCLUSIONS: Life-course and parenting experiences of young mothers are not private and located in the self, but are developed in interaction with others. Family-centered interventions are needed that support the efforts of young mothers and grandparents to become responsive caregivers. PMID- 9212511 TI - Validation of a vignette simulation of assault on nurses by patients. AB - PURPOSE: To determine the degree to which causal attribution ("blame") scores obtained from written vignettes of assault incidents, simulations of reality, reflect results that would be obtained from victims of actual assaults. DESIGN: Correlational study. Data were collected, 1990-1993, from a convenience sample of 59 RNs who had been assaulted verbally or physically at one neuropsychiatric hospital in the United States. METHODS: Victims used the Causal Attribution Scale to assign blame for their assault. Three judges then used the same scale to attribute cause for the assault based on a written description of the assault by the victim. FINDINGS: No significant differences in mean causal attribution levels were found between victims and the average ratings for the three judges for mild or severe assaults, nor between victims, judges, and the response levels obtained in two previous vignette studies. CONCLUSIONS: Mean causal attribution ("blame") scores observed in simulations that are carefully constructed assault vignettes are nearly the same as those observed in actual assaults. Vignettes appear promising as a simulation to study actual or hypothetical responses to assault. PMID- 9212512 TI - Rules outside the rules for administration of medication: a study in New South Wales, Australia. AB - PURPOSE: To improve understanding of how nurses define or redefine medication error. DESIGN: Qualitative descriptive. METHODS: This 18-week ethnomethodological study in one hospital used participant observation, documentary analysis, and validation criteria. Ethnomethodology is useful for making clearer the every-day, taken-for-granted understandings and practices of people as they make sense of their world. It hinges on the use of tacitly held knowledge in practical situations. FINDINGS: Nurses adopted practices and embodied logic to accomplish tasks. They created criteria to decide when incidents were "real errors" and used institutional rules to create order. CONCLUSIONS: The findings provide a body of tacitly held knowledge about medication error that is shared among clinical nurses and redefines medication error using six criteria. The study calls into question the way institutions seek to identify, document, and reduce medication errors by nurses and the validity of nursing research based on reported error rates. PMID- 9212513 TI - Russian Nursing Education Reform Project: new nurses for a new Russia. AB - PURPOSE: To describe a 2-year nursing exchange project, 1992-1994, designed to assess Russian nursing education and to make recommendations for change. RESULTS: Nursing education in Russia is at a level similar to vocational schools in the United States and is closely linked to medicine. Russian nurse educators were assisted in developing a philosophy of nursing, developing definitions for nursing, and incorporating process into practice. CONCLUSIONS: The Russian government and Russian nurse educators are committed to improving the quality of health care and nursing. This exchange project provided an impetus for the movement of Russian nursing education toward a process-based baccalaureate model. PMID- 9212515 TI - (Ir)reconcilable differences? The debate concerning nursing and technology. AB - PURPOSE: To review and critique the debate concerning nursing and technology. Technology has been considered both at one and at odds with nursing. ORGANIZING CONSTRUCT: Mitcham's (1994) concepts of technological optimism and romanticism. SOURCE: Nursing literature since 1960. METHODS: Historical analysis. FINDINGS: Technological optimists in nursing have viewed technology as an extension of and as readily assimilable into humanistic nursing practice, and nursing as socially advantaged by technology. Technological romantics have viewed technology as irreconcilable with nursing culture, as an expression of masculine culture, and as recirculating existing gender and social inequalities. CONCLUSIONS: Both optimists and romantics essentialize technology and nursing, treating the two as singular and fixed entities. The (ir)reconcilability of nursing and technology may be a function of how devices are used by people in different contexts, or of the (ir)reconcilability of views of technology in nursing. PMID- 9212514 TI - Peer review, authorship, ethics, and conflict of interest. AB - PURPOSE: To explore problems in peer review, authorship, ethics, and conflict of interest related to writing and publishing. Publishing and adhering to principles is critical as nurse researchers, educators, administrators, and practitioners participate in the development and dissemination of knowledge. CONCLUSIONS: The quality and integrity of nursing publications are affected by peer review, author collaboration, and ethical conduct. Understanding the conflicts of interest inherent in each action and being committed to impartial review and meeting the requirements of authorship can ensure fewer difficulties for authors, publishers, and consumers. PMID- 9212516 TI - Perspectives unifying symptom interpretation. AB - PURPOSE: To introduce the symptom interpretation model (SIM) and facilitate understanding symptoms from an intrapersonal perspective. Determining an individual's interpretation of symptoms is critical to understanding the resulting decisions. ORGANIZING CONSTRUCT: SIM is based on an illness representation model, knowledge structures theory, and propositions about reasoning. Individuals name and assign meaning to environmental stimuli. Based on this interpretation, behaviors are selected for symptom management. METHODS: Theory derivation was used to develop SIM for understanding comparisons of known and new symptoms in a behavioral outcomes context. CONCLUSIONS: Symptom familiarity reinforces patterns about symptom management. SIM enriches understanding of symptom experiences. Comprehensive assessment, including the intraindividual perspective, is essential to successful symptom management. PMID- 9212517 TI - Fathers' experience of their partners' antepartum bed rest. AB - PURPOSE: To survey paternal worries, concerns, stresses, or problems and the type of support received by men whose partners were prescribed antepartum bed rest at home, or in the hospital, or both. DESIGN: Descriptive retrospective. A national subsample of 59 men whose mates were on pregnancy bed rest were randomly selected in 1995 from a nonrandom select sample of individuals who had contacted a bed rest support group for information. METHODS: The Paternal Bed Rest Questionnaire of open-ended questions designed to detail paternal concerns, stresses, and supports was mailed to gathers. RESULTS: Major problems for fathers were assuming multiple roles, managing emotional responses, and caring for their partner. The major paternal worry was for the health of mate and fetus. Coping strategies included using tangible assistance; altering cognitive, behavioral, and emotional responses; and verbalizing worries. Fathers reported receiving little assistance from health care providers. CONCLUSIONS: Fathers experience extreme stress when pregnancy bed rest is prescribed for a mate. Family-centered care should include care of the partner whose mate is at high-risk. Interventions that reduce paternal worry and provide emotional and tangible support are needed. PMID- 9212518 TI - Linking research and practice through discussion. AB - PURPOSE: To describe the process and outcomes of a researcher-initiated discussion group with nurses in clinical practice. DESIGN: Descriptive case study. The study, which began in 1991, included one group of four acute care nurses. Between 1992-1996, three other groups of nurses participated. METHODS: Nurses in practice read and discussed articles from a program of research on breast cancer during a series of group sessions. The semi-structured sessions were tape-recorded. RESULTS: Participants suggested how to make article content more understandable to clinicians; they affirmed findings relevant to practice; and identified ways to integrate research in practice. CONCLUSIONS: Discussion served as a means for the researcher and nurses to connect the research-practice gap through learning the practice perspective and the process and value of research. Discussion groups are recommended to enhance science-based clinical practice. PMID- 9212519 TI - Cytokines: communication molecules that influence the process of disease. AB - Subset of T lymphocytes that produce different cytokine patterns have reinforced the cellular- and humoral-mediated duality of the immune response. Thus, different cytokines selectively produced by different T cells results in the Th1/Th2 T cell paradigm. These T cell subsets, in an ever widening circle of examined diseases, contribute to the resolution or persistence of a disease. Now, it is timely to examine the contribution of the cytokines produced in different disease states for the improvement of vaccines and therapies for domestic animals. PMID- 9212520 TI - Reference data on the anatomy, hematology and biochemistry of 9-month-old silver foxes. AB - Anthropometric, anatomical, hematological and biochemical reference values were estimated in clinically healthy male and female 9-month-old silver foxes. The coefficients of variation of anthropometric and anatomical measurements for 9 month-old silver foxes were as low as previously reported for adult foxes. However, in relation to body size, all measurements were smaller. Compared with adult silver foxes, higher values were observed in serum levels of triglyceride, phospholipid, beta-lipoprotein, blood urea nitrogen and total protein. Similarly, higher levels were obtained for serum enzymes, especially aspartate aminotransferase (AST), lactate dehydrogenase (LDH) and creatine kinase (CK). The high levels of these serum enzymes may be due to handling stress. Inorganic phosphorus and calcium concentrations in the young foxes were also high. The alkaline phosphatase (ALP) level, reflecting the level of bone growth, was higher than that of adults. Biochemical values of beta-lipoprotein glucose and calcium in male 9-month-old silver foxes were lower than those of females, whereas those of total cholesterol, total protein, fructosamine, iron, albumin and beta globulin were higher. PMID- 9212521 TI - Society and the not-so-new genetics: what are we afraid of? Some future predictions from a social scientist. PMID- 9212522 TI - Reassessing the influence of the Nuremberg Code on American medical ethics. PMID- 9212523 TI - Physician-assisted suicide: a tragic view. PMID- 9212524 TI - Capitation: the legal implications of using capitation to affect physician decision-making processes. PMID- 9212526 TI - A tribute to the Health Law Journal. PMID- 9212525 TI - Hazardous substance litigation in Maryland: theories of recovery and proof of causation. PMID- 9212527 TI - Personal liability implications of the duty to warn are hard pills to swallow: from Tarasoff to Hutchinson v. Patel and beyond. PMID- 9212528 TI - Putting the brakes on drive-through deliveries. PMID- 9212529 TI - Congressional action to amend Federal Rule of Evidence 702: a mischievous attempt to codify Daubert v. Merrell Dow Pharmaceuticals, Inc. PMID- 9212530 TI - Emergency! Says who?: analysis of the legal issues concerning managed care and emergency medical services. PMID- 9212531 TI - Better to lay it out on the table rather than do it behind the curtain: hospitals need to obtain consent before using newly deceased patients to teach resuscitation procedures. PMID- 9212532 TI - What is the method to their "madness?" Experimental treatment exclusions in health insurance policies. PMID- 9212533 TI - The effects of industrial employment conditions on job-related distress. AB - This paper examines the interaction between the effects of industrial unemployment and job conditions on workers' levels of psychological distress. Previous research finds that economic stress, defined as contexts of high unemployment, mainly affects distress indirectly through deteriorating job conditions. However, adaptive cost and identity salience hypotheses predict that the effects of industrial- and job-level conditions interact. I test for cross level interactions between industrial unemployment and job demands and complexity using hierarchical linear modeling, individual data for 7,095 workers from the 1987-1988 National Survey of Families and Households, and industry data from the Bureau of Labor Statistics' 1986-1988 Current Population Surveys. Economic stress at the industrial level has a direct positive effect on worker distress, and economic stress is more distressing to workers in rewarding, complex jobs. In contrast, job demands increase distress, but this effect does not interact with industrial employment conditions. PMID- 9212534 TI - The social determinants of the decline of life expectancy in Russia and eastern Europe: a lifestyle explanation. AB - This paper examines the social origins of the rise in adult mortality in Russia and selected Eastern European countries. Three explanations for this trend are considered: (1) Soviet health policy, (2) social stress, and (3) health lifestyles. The socialist states were generally characterized by a persistently poor mortality performance as part of a long-term process of deterioration, with particularly negative outcomes for the life expectancy of middle-aged, male manual workers. Soviet-style health policy was ineffective in dealing with the crisis, and stress per se does not seem to be the primary cause of the rise in mortality. Although more research is needed, the suggestion is made that poor health lifestyles--reflected especially in heavy alcohol consumption, and also in smoking, lack of exercise, and high-fat diets--are the major social determinant of the upturn in deaths. PMID- 9212535 TI - Fighting among America's youth: a risk and protective factors approach. AB - Using three nationally representative, stratified samples of youth, this paper examines health-comprising behavior (fighting) using a risk and protective factors model. In addition, the interplay between risk and protective factors is explored in an attempt to specify their precise role in acting as either mediators or buffers in the risk-taking process. Regression results indicate some similarities among the three samples and the general model's utility in predicting self-reported fighting among youth. Specifically, general environmental risk variables in the form of exposure to violence (threats and victimization) were consistent predictors of fighting among all age groups (grades 3 through 12). Upon closer inspection, analysis revealed that protective factors were not effectively mediating the relationship between risks and risk taking behavior. Results from the multiplicative regression analysis indicate that protective factors in some circumstances act as buffers; when protective factors are absent or at their weakest, the negative impact of risk on health compromising behavior was clearly present. Overall, the findings underscore the importance of taking a multidimensional approach to examining risk-taking behavior among youth where individual-level, family, school, and community factors are all considered in the design of prevention strategies. PMID- 9212536 TI - Parental support and adolescent physical health status: a latent growth-curve analysis. AB - Applying latent growth curve analysis to a sample of 310 adolescents, this study demonstrates that level of and changes in observed parental behavior are liked to the level of and changes in adolescent physical health status, respectively, through adolescent perception of parental support. In addition, the level of observed parental behavior had a significant direct effect on subsequent changes in adolescent health status. The results provide evidence for the influence of parental support on adolescent physical health, both directly and indirectly through the adolescent's perception of that support. Confidence in the findings is strengthened by (1) employing a prospective, longitudinal research design, (2) analyzing intraindividual changes in support and health, and (3) reducing potential method variance confounds by using multi-informant reports of parental behavior. PMID- 9212537 TI - The continuation of family caregiving in Japan. AB - The purpose of this study was to develop concepts that facilitate our understanding of why family caregivers of demented elderly persons can continue caregiving despite various difficulties of care. Twenty-six Japanese daughter or daughter-in-law caregivers of elderly parents with dementia who lived at home or in long-term care facilities were recruited through various senior service organizations in Japan. The caregivers underwent unstructured interviews, and the interview data were analyzed using the constant comparative method. Three categories emerged as reasons for care continuation: value of care, maintainers of value, and reinforcers of care continuation. Value of care came from societal norms and attachment, and was the basis of caregivers' motivation to continue care. Several maintainers of value and reinforcers of care continuation also emerged from the analysis. The contents and some longitudinal changes in these categories were explained. The findings highlight the need to assess these categories separately in order to develop appropriate interventions and they have implications for future research and policy development. PMID- 9212538 TI - On stigma and its consequences: evidence from a longitudinal study of men with dual diagnoses of mental illness and substance abuse. AB - Numerous studies have demonstrated a strong connection between the experience of stigma and the well-being of the stigmatized. But in the area of mental illness there has been controversy surrounding the magnitude and duration of the effects of labeling and stigma. One of the arguments that has been used to downplay the importance of these factors is the substantial body of evidence suggesting that labeling leads to positive effects through mental health treatment. However, as Rosenfield (1997) points out, labeling can simultaneously induce both positive consequences through treatment and negative consequences through stigma. In this study we test whether stigma has enduring effects on well-being by interviewing 84 men with dual diagnoses of mental disorder and substance abuse at two points in time--at entry into treatment, when they were addicted to drugs and had many psychiatric symptoms and then again after a year of treatment, when they were far less symptomatic and largely drug- and alcohol-free. We found a relatively strong and enduring effect of stigma on well-being. This finding indicates that stigma continues to complicate the lives of the stigmatized even as treatment improves their symptoms and functioning. It follows that if health professionals want to maximize the well-being of the people they treat, they must address stigma as a separate and important factor in its own right. PMID- 9212539 TI - Speeding up the improvement process. PMID- 9212540 TI - Automation: keeping it all together. PMID- 9212541 TI - Chemung Valley Health Network: the development of a community health information network. PMID- 9212542 TI - Patient satisfaction with nurse practitioner care in primary care. AB - This quantitative, descriptive pilot study assessed patient satisfaction with care provided by four nurse practitioners using a modified version of the Di Tomasso-Willard Patient Satisfaction Questionnaire. Results indicated high satisfaction with care in all groups, but there were differences among the groups on 26 percent of the items and on three of the five subscales. The implication is that nurse practitioners need to identify and improve those dimensions of care for which patients are less satisfied. PMID- 9212543 TI - The relationship of patients' perceptions of holistic nurse caring to satisfaction with nursing care. AB - Hospitalized patients interact with nurses more often than with other health care providers. Qualitative studies have identified patients' and nurses' perceptions of nurse caring behaviors. Few studies have explored nurse caring behavior in relation to patient satisfaction. The article describes a study designed to determine whether there is a relationship between patients' perceptions of nurse caring and satisfaction with nursing care and whether selected patient variables significantly affect that relationship. In the patient population studied, after controlling for patient variables of age, gender, and level of pain, a significant positive relationship was found between patients' perceptions of nurse caring and their satisfaction with nursing care. PMID- 9212544 TI - Outcomes in an academic nursing center: client satisfaction with student services. AB - An important element of quality assessment/quality improvement efforts in health care organizations is the evaluation of patient satisfaction. This element is also important in academic nursing centers, where students provide home visiting services to improve access to care for vulnerable populations in the community. Using a modified version of an existing patient satisfaction instrument, faculty and staff of the University of North Dakota Nursing Center surveyed clients at the end of each semester of service. Results indicated overwhelming satisfaction with all aspects of care provided by the students. Other effects of the program on clients are discussed. PMID- 9212545 TI - First impressions of the nurse and nursing care. AB - Patients (N = 1,180), nurses (N = 918), and administrators (N = 332) in 22 acute care hospitals across the country were surveyed regarding their first impression of the professional image communicated by nurses' uniforms. The Nurse Image Scale, with pictures of the same nurse in nine different uniforms, was used as the data gathering tool. A comparison of the mean score of each uniform as rated by all respondents (N = 2,430) showed the white pant uniform with stethoscope was rated significantly higher than other uniforms. The white pant uniform with cap, dress with cap, pants suit, and dress with stethoscope scored closely in a second place grouping. The white dress uniform and street clothes with laboratory coat tied for third place. Colored designer scrubs and white pants with colored top scored lowest. Ratings of patients, nurses, and administrators were similar, although patients tended to rank some uniforms significantly differently than nurses and administrators. The nurse in the pant uniform with stethoscope was most preferred for care. Least preferred was the nurse in colored scrubs and street clothes with lab coat. These findings point to the need for nurses to be differentiated from auxiliary health care personnel and to project a professional image in a competitive health care environment. PMID- 9212546 TI - Standards and practice guidelines as the foundation for clinical practice. AB - The structure and organization of health care delivery are in the midst of rapid change. Health care providers from a variety of disciplines are being challenged to define their practice and the expected patient outcomes resulting from their processes of care delivery. Standards and clinical practice guidelines are important tools for enhancing the quality of health care delivery and for documenting care. The article describes a process for developing standards and clinical practice guidelines and presents an organizational scheme for them. Based on recommendations from diverse national groups, a format for practice guidelines is presented, and a system for implementation and ongoing evaluation is recommended. PMID- 9212547 TI - Care management: outcomes-based practice for the primary nurse. AB - Managed care penetration is increasingly dramatically in most health care markets across the country. The impact is the radical redesign of health care systems and the reshaping of the approach to health care from an illness model to a wellness, health promotion model. Integrated delivery systems will facilitate the patient's journey through the continuum of care. The article describes how one institution is streamlining that journey through the integration of care management into the role of the primary nurse who provides direct care. PMID- 9212548 TI - Supplemental stimulation of premature infants: a treatment model. AB - Examined the human infant literature on supplemental stimulation to delineate a course of intervention based on the ontogeny of the nervous system and the impact that systematic stimulation may have on behavioral organization in the premature infant. Effects of vestibular, tactile/kinesthetic, auditory, and oral stimulation are discussed with respect to their similarity to the intra- or extra uterine environment. Long-standing theoretical and methodological problems are discussed, and a "sequential multimodal treatment model" is introduced. PMID- 9212549 TI - Low birth weight and parenting stress during early childhood. AB - Identified factors associated with parenting stress among the parents of low birth weight children participating in a prospective longitudinal study. The child's development status and the quality of the infant-parent relationship contributed to early childhood parenting stress beyond the contribution of neonatal medical risk, which was a significant predictor until the concurrent measure of child development status was entered in the regression analysis. Results are consistent with a model of processes underlying the experiences of parents of medically vulnerable infants presented by the Goldberg and Marcovitch (1986) model. Replication of these findings among parents of specific subgroups of low birth weight children represents a direction for future research. PMID- 9212550 TI - Revision of a parent-completed development screening tool: Ages and Stages Questionnaires. AB - Examined the Ages and Stages Questionnaires (ASQ), a series of 11 developmental questionnaires designed to be completed by parents and caregivers of young children from 4 to 48 months of age. The ASQ were recently revised and additional psychometric data were gathered. Analyses on over 7,000 questionnaires indicated high test-retest reliability, interobserver reliability, and internal consistency. Concurrent validity using standardized measures yielded on overall agreement of 85%, with a range of 76-91%. Specificity was high across questionnaire intervals while sensitivity was lower and varied across intervals. Use of parent-completed screening tools such as the ASQ is attractive in terms of cost-effectiveness, parental involvement, and flexibility in administration procedures. PMID- 9212551 TI - Mothers' beliefs about the causes of infant growth deficiency: is there attributional bias? AB - Tested for defensive attributional bias in mothers' causal explanations for infant (2-12.5 months) growth deficiency. Mothers of healthy babies (controls; n = 82), growth deficient babies without medical problems (n = 27) and growth deficient babies with mild medical problems (n = 22) rated their levels of agreement with 23 causes of growth problems which were designed to vary in the degree of personal threat to parenting self-esteem. Ratings were completed for the mother's (Own) baby and for a nonspecific (Other) baby. Findings partially support a theory of defensive attributional bias, with higher agreement when causes referred to Other (vs. Own) baby, and lower agreement with family-related than with medical/nutritional causes. Factors that may have influenced material experience of threat and implications of the findings for clinical practice are discussed. PMID- 9212552 TI - The use of scheduled awakenings to eliminate childhood sleepwalking. AB - Evaluated the use of scheduled awakenings to eliminate sleepwalking using a noncurrent multiple baseline design across three subjects with persistent sleep walking. Treatment procedure involves having parents awaken children several hours after they go to sleep and just before the typical time of the sleepwalking episode. This intervention proved immediately successful in eliminating sleep walking in all three children. Treatment effects were maintained at 3 and 6 months posttreatment. Implications for intervention and future research are discussed. PMID- 9212553 TI - Nurse coaching and cartoon distraction: an effective and practical intervention to reduce child, parent, and nurse distress during immunizations. AB - Evaluated a low cost and practical intervention designed to decrease children's, parents', and nurses' distress during children's immunizations. The intervention consisted of children viewing a popular cartoon movie and being coached by nurses and parents to attend to the movie. Ninety-two children, 4-6 years of age, and their parents were alternatively assigned to either a nurse coach intervention, a nurse coach plus train parent and child intervention, or a standard medical care condition. Based on previous findings of generalization of adult behaviors during medical procedures, it was hypothesized that training only the nurses to coach the children would cost-effectively reduce all participants levels of distress. Observational measures and subjective ratings were used to assess the following dependent variables: children's coping, distress, pain, and need for restraint; nurses' and parents' coaching behavior; and parents' and nurses' distress. Results indicate that, in the two intervention conditions, children coped more and were less distressed, nurses and parents exhibited more coping promoting behavior and less distress promoting behavior, and parents and nurses were less distressed than in the control condition. Although neither intervention was superior on any of the variables assessed in the study, nurse coach was markedly more practical and cost-effective. Therefore, nurses' coaching of children to watch cartoon movies has great potential for dissemination in pediatric settings. PMID- 9212554 TI - Psychosocial functioning of children, adolescents, and adults following hypospadias surgery: a comparative study. AB - Used standardized questionnaires to compare psychosocial functioning of 116 children and adolescents (9 to 18 years) and 73 adults (18 to 38 years) operated on for hypospadias, a congenital penile anomaly, with that of 88 and 50 age matched comparison males, respectively, treated for an inguinal hernia. The relationships of coping with penile appearance, subject age, severity of hypospadias, number of operations, age at final surgery, and type of surgical procedure with psychosocial functioning were also investigated. Hypospadias patients did not exhibit a poorer psychosocial functioning and no significant relationships of various medical characteristics with psychosocial functioning could be discerned. Genital/body perception of hypospadias patients ages 9 to 18 years correlated positively with psychosocial functioning, albeit with low values. These findings are important for psychologists and specialists in the counseling process of hypospadias patients and their parents. PMID- 9212555 TI - Sleep patterns among children with attention-deficit hyperactivity disorder: a reexamination of parent perceptions. AB - Surveyed parents of children with and without ADHD for their perceptions of their children's sleep patterns. All children had been referred for learning or behavior problems to an outpatient assessment center. Diagnoses of ADHD were based on DSM-III-R, rather than DSM-III criteria, avoiding a possible confound from diagnostic criteria that formerly included sleep disturbance as a defining characteristic of ADHD. Data replicated past findings showing that parents perceive children with ADHD to have greater sleep difficulty than normally developing children. Parents perceived few differences between sleep patterns of children with ADHD who were taking or not taking stimulant medication. Implications of these findings are discussed in the context of past literature and present clinical practice. PMID- 9212556 TI - The development and validation of the Children's Hope Scale. AB - Assuming that children are goal-oriented, it is suggested that their thoughts are related to two components--agency and pathways. Agency thoughts reflect the perception that children can initiate and sustain action toward a desired goal; pathways thoughts reflect the children's perceived capability to produce routes to those goals. Hope reflects the combination of agentic and pathways thinking toward goals. A six-item dispositional self-report index called the Children's Hope Scale is introduced and validated for use with children ages 8-16. Results suggest that the scale evidence internal consistency, and is relatively stable over retesting. Additionally, the scale exhibits convergent, discriminant, and incremental validity. Limitations and uses of the scale are discussed. PMID- 9212557 TI - Noise pollution. PMID- 9212558 TI - Bull horn injuries in rural India. AB - Bull horn injuries though not so commonly seen in cities, are commonly observed in rural areas. The bull horn injuries are different in many ways from other casualties e.g., blunt injuries, stab injuries, road traffic accidents, etc. A total of 50 cases of bull horn injuries were managed at a Rural Medical College during a 5-year period. An emphasis is given on the nature and mechanism of these types of injuries and the measures to prevent such mishaps. PMID- 9212559 TI - Spectrum of benign breast disorders in a university hospital. AB - In a period of 2 years, 234 cases of benign breast disorder were studied. Breast pain and modularity was the commonest group (70.1%) followed by fibroadenoma (17.5%). Cyclical mastalgia (61.5%) is more common than non-cyclical mastalgia (38.5%). The age of the patients with cyclical mastalgia was significantly lesser than patients with non-cyclical mastalgia. Cyclical mastalgia was seen only in premenopausal females while non-cyclical mastalgia was also seen in postmenopausal females. Treatment with vitamin E showed 41% response rate with minimal side-effects while treatment with danazol showed 72.1% response rate but was associated with side-effects in one third of the patients. PMID- 9212560 TI - CSF/blood glucose ratio and other prognostic indices in pyogenic meningitis. AB - Fifty cases of pyogenic meningitis were examined for various prognostic indices, especially cerebrospinal fluid (CSF)/blood glucose ratio. Overall mortality was 40%. Age below one year and depressed level of consciousness were associated with high mortality. Illness of more than 7 days, presence of associated illness and absence of neck rigidity were not found to be statistically significant factors associated with higher mortality. CSF leucocyte count of more than 1000 cells/cmm and CSF protein more than 500 mg/dl were statistically significant factors associated with higher mortality. In cases of CSF glucose level below 20 mg/dl and CSF/blood glucose ratio below 0.2, the increase in mortality was highly significant. CSF/blood glucose ratio in cases who recovered was much higher than those who died. CSF/blood glucose ratio increased to normal in cases who recovered but remained low in cases who expired. PMID- 9212561 TI - Approach to aerosol therapy in management of asthma at primary care level. AB - Present day management of bronchial asthma focuses on use of inhaled drugs. To find out the acceptability of aerosol therapy at primary care level, a questionnaire survey was conducted on practical acceptance of aerosol therapy in management of asthma. Seven clinically relevant questions were asked regarding management modalities followed in bronchial asthma. Amongst the group of general practitioners (n = 60) studied, it was found that in treatment of chronic stable asthma inhaled steroids are used by only 50% and inhaled bronchodilators by 86.7%, whereas 93.3% use oral bronchodilators and 60% use oral steroids to treat the same. During exacerbation injectable bronchodilators are preferred to inhaled drugs. Poor compliance to aerosol therapy was reported by 33.3%. Reasons for non compliance are discussed, important ones are cost and technique. Spacer was reported to be useful by 20% and lung functions are carried out occasionally by only 5% of practitioners. PMID- 9212562 TI - HIV infection in obstetrics and gynaecology. PMID- 9212563 TI - Strategy for the development of emergency medical services in India. PMID- 9212564 TI - Retirement planning for doctors--the easy way. PMID- 9212565 TI - Comparative evaluation of drug advertisements in Indian, British and American medical journals. PMID- 9212566 TI - Primary amenorrhoea out of craniopharyngioma. PMID- 9212567 TI - Consecutive five abortions in a case of septate uterus--ended in a term pregnancy following modified Jones' metroplasty operation. PMID- 9212568 TI - A case of renal artery stenosis complicating pregnancy. PMID- 9212569 TI - Malignant Brenner tumour of ovary. PMID- 9212570 TI - Crisis in health care for the people. PMID- 9212571 TI - Association of dietary ghee intake with coronary heart disease and risk factor prevalence in rural males. AB - To determine the association between intake of dietary fat, specifically Indian ghee, and prevalence of coronary heart disease (CHD) and risk factors as study was undertaken on a rural population in Rajasthan. Total community cross sectional survey was done using a physician administered questionnaire; 1982 males aged 20 years and more were studied. The dietary questionnaire focused on the amount and type of fat consumed. Staple dietary fat in this area is mustard/rapeseed oil and Indian ghee. To define the role of ghee, the average amount consumed in a month was determined; 782 males (39%) consumed 1 kg or more ghee per month (group 1) and 1200 (61%) consumed less than 1 kg per month (group 2). To elicit details of fatty acid composition of the diet consumed, detailed dietary history was acquired from a random 460 (23%) males; 220 from group 1 and 240 from group 2. Group 1 males were significantly younger, more literate and had more weight and body-mass index. This group consumed significantly more calories, saturated and mono-unsaturated fats while the consumption of polyunsaturated fats was similar in the two groups. Fatty acid intake analysis showed that group 1 males consumed more mono-unsaturated (n-9) fatty acids than group 2. Intake of polyunsaturated n-3 and n-6 fatty acids was similar. There was significantly lower prevalence of CHD in men who consumed > kg ghee per month (odds ratio = 0.23, 95% confidence limits 0.18-0.30, p < 0.001). Multivariate analysis confirmed this association (p < 0.001). The prevalence of hypertension and other coronary risk factors was similar in the two groups. PMID- 9212572 TI - A simple risk estimates study for oral cavity cancer: practical approach in Indian context. AB - A study was conducted on 131 cases of oral cavity cancer (OCC), 145 cases of oral leucoplakia and 704 subjects without any oral lesions to investigate risk factors associated with the development of carcinoma of oral cavity in a hospital based cancer registry. Personal interviews, as well as physical examinations of the subjects enabled evaluation of a variety of potential risk factors. Potential risk factors like tobacco chewing, tobacco smoking, snuff dipping, alcohol consumption, bad oral and dental hygiene and age were given each certain numerical values. Each subject was first given a scoring and then analysed and correlated with the presenting lesions, when present. The study revealed that tobacco chewing and bad oral and dental hygiene contributed mainly to higher scoring. Among the subjects in high risk group (scoring more than 400) 63% had OCC, 21% had oral leucoplakia and 16% had no clinical oral lesions. Among the medium risk group (scoring between 100 and 400) 6% had OCC, 21% had leucoplakia and 73% had no oral lesions. In low risk group (scoring below 100) 8% had leucoplakia and 92% had no clinical oral lesions. Using the scoring system, it is suggested that the high risk group for OCC could be identified from general population and cancer detection tests could be specially directed towards this target group to detect maximum number of cases with minimum possible resources. PMID- 9212573 TI - Reservoirs of nosocomial pathogens in neonatal intensive care unit. AB - A total of 256 swabs taken from different areas of neonatal intensive care units (ICU) in KCG Hospital and AMC Hospital, Bangalore were bacteriologically investigated for prevalence, source and spread of nosocomial bacteria. Culture studies revealed growth in 217 (84.8%) swab samples indicating considerable contamination of different areas of the units and sources of infection. Klebsiella pneumoniae (27.3%) was the predominant organism followed by Esch coli (16.8%), Staph aureus (11.7%), Staph epidermidis and Pseudomonas aeruginosa (10.2%), enterococcus and proteus (4.7%), Citrobacter freundi (3.5%) and Clostridium tetani (2.4%) isolated from the equipment, cradles, other inanimate objects and environmental surfaces. Out of 312 isolates, monobacterial prevalence was 43.6% in contrast to polybacterial prevalence of 56.4%. Klebsiella pneumoniae (74.3%) was the predominant monobacterial isolate. The indoor air of the units was found to carry common nosocomial bacteria of 4 or more different bacterial species at dangerous levels as observed by colony counts of 15 to 30 on exposed blood agar plates. Almost all sources in ICU revealed the presence of Klebsiella Pneumoniae, Esch coli, pseudomonas and staphylococcus thus forming the potential reservoirs of nosocomial infections to babies and this could be attributed to overcrowding, poor ventilation system and failure to follow basic principles of strict protective barrier nursing. PMID- 9212574 TI - Knowledge and attitude in relation to HIV/AIDS among in-service nurses of Calcutta. AB - Seventy-five senior nurses attending a workshop were surveyed with questionnaires and using two separate scales, their knowledge about transmission and precautionary measures, and their general attitude towards HIV/AIDS as well as willingness for patient-care were assessed. The nurses showed a satisfactory level of knowledge (mean percentage score 74.3), but misconceptions regarding disinfection and precautionary measures were present; 33% had overall negative attitudes and 24% unwilling to provide care for HIV-infected patients. Knowledge and attitude were positively correlated (r = .32). Knowledge deficits of some aspects of infection leading to fear of contagion and judgemental outlook towards HIV infection might lead to negative attitude impeding proper care. It is suggested that continuous in-service training be instituted to dispel misconceptions and to develop favourable and non-discriminatory attitude. PMID- 9212576 TI - Psychosis during disulfiram therapy for alcoholism. AB - Fifty-two patients (51 males, one female) of alcohol dependence/abuse diagnosed according to DSM 111 R1 were registered for disulfiram treatment and were admitted for a 4-week period. No alcohol challenge test was given during their hospital stay. Disulfiram tablets were administered 250 mg twice daily after food. After one week's hospital stay with disulfiram therapy they were sent home to be followed up initially after 15 days and later on once a month. Six male patients developed psychotic symptoms. All of them had shown a mood disorder but no thought disorder was observed. Psychotic symptoms remitted completely after withdrawing disulfiram and then a short course of antipsychotic therapy was given, except in one patient who had to be given lithium for remission of symptoms. PMID- 9212575 TI - A study of perinatal mortality and associated maternal profile in a medical college hospital. AB - Study of perinatal mortality in a Medical College Hospital revealed stillbirth rate as 38.4, early neonatal death rate as 29.3 and overall perinatal morality rate as 67.7 per 1000 live births. More than half (53.6%) of the perinatal deaths were in primipara and another 22.8% in mothers of parity more than 3. Most mothers (85.9%) did not receive adequate antenatal care services. On admission 35.1% mothers presented with some risk factors. The major risk factors identified were toxaemia of pregnancy (14.8%), severe anaemia (13%) and antepartum haemorrhage (2.6%). PMID- 9212577 TI - Cough--a common complication of ACE-inhibitor therapy. PMID- 9212578 TI - Subdural empyema due to external burns. PMID- 9212579 TI - Splenic hydatid cyst in children--a report of two cases. PMID- 9212580 TI - Unusual presentations of typhoid fever. PMID- 9212581 TI - Mature cystic teratoma of the fallopian tube associated with ectopic pregnancy. PMID- 9212582 TI - Extravasation of urine following forced diuresis. PMID- 9212583 TI - Peri-urethral abscess--a rare presentation of urethral carcinoma. PMID- 9212584 TI - Medical ethics. PMID- 9212586 TI - Fragile X syndrome in India. PMID- 9212585 TI - Tuberculosis in the skin. PMID- 9212587 TI - Neurotrophic activity in cytokine-activated astrocytes. AB - Accumulating evidence indicates that various neurotrophic factors (NTFs) exist and function in the brain. In the mature mammalian brain, NTF expression is exclusively restricted to neurons. However, astrocytes activated by various cytokines, including fibroblast growth factor and interleukin-1 beta, produce a significant amount of nerve growth factor (NGF) in vitro. Furthermore, non-NGF type NTF expression in astrocytes is also activated by the cytokines. The cytokines also enhance both release of ciliary neurotrophic factor from and expression of high-molecular weight basic fibroblast growth factor (FGF) in astrocytes. In the early phase following brain injury, cytokine-activated astrocytes rescue the damaged neurons via NTFs and other biologically active molecules. PMID- 9212589 TI - Management of poor-grade patients with ruptured intracranial aneurysm. AB - To formulate treatment strategies for poor-grade patients after aneurysmal subarachnoid hemorrhage (SAH), medical records were analyzed for 166 patients who were in Hunt and Hess Grade IV or V among 588 consecutive cases with ruptured intracranial aneurysm admitted during the past 5 years. Causes of unfavorable outcome (poor or dead) in those 166 patients were evaluated to improve the management outcome. Overall management results of the 166 poor-grade patients were favorable (good or fair) in 71 (42.8%), unfavorable in 95 (78 dead, 17 poor). Direct clipping was performed in 90 patients, and the results were favorable in 69 (76.7%) and unfavorable in 21 (23.3%). Surgery was not done in 76 patients because 41 were moribund on arrival, 15 deterioration due to rebleeding, 7 severe brain swelling, 5 serious medical illness, one severe delayed ischemic deficit (DID), and one cerebral infarction following angiography, and 6 refused surgery. Seven patients survived in non-surgery group (2 fair, 5 poor). Direct effects of aneurysm rupture (34.8%) and early rebleeding (34.8%) were the major causes of unfavorable outcome in Grade IV patients, while it was direct effect of aneurysm rupture (91.8%) in Grade V patients. It is suggested that as rebleeding is the only preventable cause of unfavorable outcome, urgent management is necessary to prevent rebleeding, especially for Grade IV patients. Grade IV patients should be treated aggressively with direct clipping for non-complex aneurysms or for patients with hematoma, and coil embolization for complex aneurysms without hematoma. PMID- 9212588 TI - Angiotensin II signal transduction in vascular smooth muscle cells: role of tyrosine kinases. AB - Originally known to be a vasoconstrictor and thought to play a critical role in hypertension, angiotensin II has recently emerged to be important in inflammation, atherosclerosis and congestive heart failure. The expanding role of angiotensin II implies that multiple signal transduction pathways are likely to be activated in a tissue-specific manner. Recent data show that angiotensin II stimulates not only cytoplasmic tyrosine kinases including c-Src, focal adhesion kinase (FAK), and Janus kinases (JAK2 and TYK2), but also may transactivate receptor tyrosine kinases such as Axl and PDGF by as yet undefined autocrine/paracrine mechanisms. Finally, tyrosine kinases, which mediate tyrosine phosphorylation of key signal mediators such as Shc, Raf, and phospholipase C gamma following angiotensin II stimulation, remain to be defined. These tyrosine kinases, activated by angiotensin II, appear to be required for angiotensin II effects such as vasoconstriction, proto-oncogene expression, protein synthesis, and cell proliferation. Thus, it is important to understand angiotensin II mediated signaling events, especially those related to tyrosine kinase activity, to develop new therapies for cardiovascular diseases. PMID- 9212590 TI - Disappearance of serum hepatitis C virus RNA within two days after one dose interferon administration is predictive for response to high-dose interferon alpha 2b treatment for chronic hepatitis C. Keio Interferon-alpha 2b Study Group. AB - We have reported the Keio multicenter randomized trial of interferon-alpha 2b treatment for chronic hepatitis C, hypothesizing that disappearance of serum hepatitis C virus (HCV) ribonucleic acid (RNA) during the first 2 days by one does administration of interferon is a predictive factor of final response to the high-dose interferon treatment. In this study we quantified HCV RNA by multicyclic reverse transcription-polymerase chain reaction in the stored sera of the same patients with our previous study. The multivariate analysis confirmed that the pretreatment HCV RNA levels and HCV genotype were significantly correlated with the response to the 6-month course interferon treatment. Although the relationship between decreased HCV RNA titers within the first 2 days after one dose administration of interferon and the efficacy of the therapy was not obtained, the cases in which HCV RNA disappeared within 2 days significantly responded to the 6-month course treatment (73.7% vs 12.5% in other cases, p < 0.01). The present study has confirmed the hypothesis suggested in the previous study that clearance of HCV RNA during the first 2 days has a predictive value for the final outcome. PMID- 9212591 TI - Superacute phase blood pressure elevation may relate to massive hematoma in hypertensive putaminal hemorrhage. AB - A restrospective clinical investigation has been performed to elucidate the relationship between hematoma size in putaminal hemorrhage and blood pressure (BP) changes during the immediate post-hemorrhagic phase in the emergency room (ER). Thirty-seven adult patients brought to the emergency department by ambulance within 6 hours after onset of symptoms with a confirmed diagnosis of acute putaminal hemorrhage on CT have been involved. Two BP measurements during the superacute phase in the ER have been studied: immediately after arrival at the ER (BP-I), and immediately prior to CT examination (BP-II). Patients have been divided into 6 categories: 1) those whose BP decreased with treatment (D+), 2) those whose BP decreased without treatment (D-), 3) those whose BP increased in spite of treatment (I+), 4) those whose BP increased without treatment (I-), 5) those whose BP remained unchanged in spite of treatment (U+), and 6) those whose BP remained unchanged without treatment (U-). Hematoma size has been compared among 5 categories (D+, D-, I-, U+, U-) using factorial ANOVA (analysis of variance). The hematoma sizes have been found to be (D+) 54 +/- 44 ml, (D-) 22 +/- 25 ml, (I-) 102 +/- 58 ml, (U+) 11 +/- 5 ml, (U-) 21 +/- 9 ml (mean +/- S.D.), respectively. (I-) has been significantly larger than any of the other categories (p < 0.001 - 0.05). Additional ANOVA has shown that BP-II in category (I-) was significantly higher than that of the other categories. Patients with putaminal hemorrhage whose BP was elevating during the superacute phase in the ER were shown to have massive hematomas. PMID- 9212592 TI - Rapid reduction in ryanodine binding of hippocampus CA1 in cerebral ischemia. AB - Ryanodine receptors located on the sarcoplasmic or endoplasmic reticulum, play an important role in the regulation of the intracellular Ca2+ level via the mechanism of Ca(2+)-induced Ca2+ release (CICR). Perturbation of intracellular Ca2+ regulation has been considered to be one of the most important mechanisms underlying acute ischemic neuronal damage. The ryanodine binding, an indicator of intracellular channels of CICR, and local cerebral blood flow (LCBF) were therefore examined at 15 min post-ischemia in the gerbil brain. The autoradiographic method developed in our laboratory enabled us to determine both parameters within the same brain. Severe hemispheric cerebral ischemia was induced by occluding the right common carotid artery. LCBF was measured at the end of the experiment using [14C]iodoantipyrine method. The ryanodine binding was evaluated autoradiographically in vitro using [3H] ryanodine. A group of gerbils who underwent a sham procedure served as controls. LCBF was found to be significantly decreased in most cerebral regions on the occluded side. In contrast, a significant reduction in ryanodine binding was noted only in the hippocampus CA1 on the occluded side. Taken together, these findings indicate that the CICR in the hippocampus CA1 may be especially susceptible to acute ischemic stress, and be closely associated with the pathophysiological mechanisms of the selective vulnerability of this region. PMID- 9212593 TI - A female with aneurysm of aortic arch due to lung cancer. PMID- 9212595 TI - Modeling the lifespan of human T lymphocyte subsets. AB - T lymphocytes may be classified as naive or memory cells, depending on whether they possess immunological memory. The central tenet of this work is that memory lymphocytes revert to naive lymphocytes in vivo. This phenomenon is modeled by a mechanism qualitatively similar to Demoivre's law of human mortality. Additionally, both lymphocyte subsets undergo decay. Model parameters and their standard errors are estimated by maximizing a likelihood function constructed by Kalman filtering of the experimental data on the assumption that it is contaminated by measurement noise of constant relative error. PMID- 9212594 TI - Steel's potential doubling time and its estimation in cell populations affected by nonuniform cell loss. AB - The in vivo infusion of the thymidine analogue bromodeoxyuridine (BrdUrd). followed by delayed biopsy and bivariate DNA-BrdUrd flow cytometry, makes it possible to estimate Steel's potential doubling time (Tpot) of human tumors. In the present paper, the expression of Steel's Tpot for a rather general cell population model, in which the distribution of cell loss is assumed to be nonuniform, is derived in terms of the model parameters. We show that Steel's Tpot of a population can be markedly different for the doubling time that would be exhibited by the population in the absence of cell loss. These doubling times, on the contrary, are equal when loss is uniform. Moreover, we studied the influence of modes of cell loss different from the uniform random loss on the estimation of Tpot by using the labeling index or the nu-function, quantities that can be determined from the bivariate DNA-BrdUrd distribution. PMID- 9212596 TI - Estimating clonal heterogeneity and interexperiment variability with the bifurcating autoregressive model for cell lineage data. AB - We utilize an extension of the variance-components models for cell lineage data in Huggins and Staudte (R.M. Huggins and R.G. Staudte, Variance components models for dependent cell populations. J. Am. Stat. Assoc. 89:19-29 (1994) to analyze NIH3T3 cells grown in two different media. This modeling approach has the advantage of a simple built-in correlation structure between familial members and allows for estimating experimental effects, rather than treating them as random effects. In addition, this methodology gives robust estimates of model parameters together with standard errors required for statistical inference. The importance of clonal heterogeneity and interexperiment variability in modeling eukaryotic cell cycles was previously pointed out by Kuczek and Axelrod (T. Kuczek and D.E. Axelrod, The importance of clonal heterogeneity and interexperimental variability in modeling the eukaryotic cell cycle. Math. Biosci. 79:87-96 (1986). This analysis confirms significantly positive sister-sister correlation when cells are grown in rich or poor medium and negative mother-daughter correlation when cells are grown in poor medium. However, for cells grown in rich medium, Kuczek and Axelrod's analysis gives negative mother-daughter correlations, whereas this analysis gives significant positive mother-daughter correlations. PMID- 9212598 TI - Intellectual property law in biotechnology. PMID- 9212597 TI - Anticipatory decision making & incompetent patients. Recent developments in Europe. PMID- 9212599 TI - De facto gatekeeping and informed consent in intensive care. AB - Medical decision-making is based on the doctrine of informed consent which is, in turn, based on autonomy, which represents one of four pillars of medical ethics, the others being beneficence, non-malfeasance and social justice. Decision-making in intensive care with respect to the withdrawal of treatment, in particular ventilator therapy, is often extremely difficult for patients or their relatives and they would rather not make any decision other than to insist on the maintenance of therapy in spite of sound, reasonable medical advice that such therapy is of no value to the patient. Aside from issues of a dignified death, this is likely to be to the detriment of other patients who might be refused admission to intensive care and thus is counter to the dictates of social justice. Under these circumstances, there would appear to be a need to give authority to the reasonable medical decision to discontinue resuscitation. PMID- 9212600 TI - Practice guidelines & medical malpractice litigation. AB - In 1973, the United States Congress enacted legislation requiring physicians to initiate Peer Review Organizations to monitor utilization and quality of hospital and physician services in the federally funded Medicare program. A hardly noticed provision of the statute intimated the desirability of formulating guidelines for medical treatment. What was originally intended to simplify and universalize general standards by which quality of care could be objectively measured has more recently escalated into formalized projects, subsidized by government, to create "practice parameters". The impetus to define clinical conditions and methods of treatment for specific medical conditions (practice parameters) and standards of practice to avoid or defend malpractice claims (risk management protocols) are part of the movement in the United States for tort reform. If the vague "reasonable man" standard of care in negligence law can be supplanted by a scientifically developed, particularized medical practice standard, it is anticipated that spurious claims and defensive medical practice will be discouraged, quality improved, iatrogenic injury and malpractice litigation diminished. Many U.S. states undertook tort reform in the last decade. A few have embarked on medical-legal reform. One state is conducting a five-year medical liability project that calls for the development of practice parameters and risk management protocols in four medical specialties. The parameters will have the effect of law and may be introduced as evidence in medical malpractice trials. How the parameters are established, their effect on the strategies of litigation, the resultant trial problems in the introduction of evidence and in the burden of proof and their potential for acceptance by a significant number of jurisdictions are the issues to be explored in this paper. PMID- 9212601 TI - Survey of the understanding and expectations of patients referred for observation. PMID- 9212602 TI - To inform or not to inform--a decision with psychobiological implication. AB - A patient, being informed about his dangerous or even fatal disease, like cancer, has a higher chance to give up and display a renunciation of search activity. Such renunciation decreases body resistance and makes the overall situation hopeless. Thus a person has the right to be not informed about such diseases, because he has the right to use all his inner resources for survival. PMID- 9212603 TI - Myth and reality of informed consent in clinical trials. AB - Informed consent is a prerequisite for any medical intervention or for participation in scientific research. Legal requirements in health care towards informed consent provide qualifications of information disclosure and focus on procedures to obtain a valid consent. The premise of informed consent as a rational decision making process implies an informed consideration, assessment and patient choice and may be perceived as an ideal in the nature of a myth. These principles and assumptions will be discussed to obtain more insight into informed consent in clinical trials. A cogent model of informed consent is presented, which could be useful in clinical trials. We suggest a gradual distinction between informed consent in regular treatment vs. clinical trials, based on concepts like achievable benefits and risk disclosure. Suggestions are made for further empirical research, to obtain more insight into the myth and reality of informed consent in clinical trials. PMID- 9212604 TI - Inter-examiner variability. PMID- 9212605 TI - Mental disorders in abnormal offenders in Papua New Guinea. AB - The case notes of all 64 referred abnormal offenders (mental patients with criminal records) sent to a psychiatric hospital between January 1971 to May 1996 were examined. It was found that severe mental disorder like schizophrenia (27 out of 64) was the most common cause of violent crimes such as homicide. Epilepsy 10.9 (n = 7) was another important neuropsychiatric condition related to violence. Alcohol and cannabis abuse were an associated factor in 21 (32.8%) referred cases. Culture bound syndromes like "Amok Syndrome" and "Spirit Possession Syndrome" were also found as a cause of violent behavior. PMID- 9212606 TI - The views of psychiatric patients and their treating physicians of court-ordered compulsory hospitalization for criminal acts. AB - The legal responsibility for the mentally ill has long been a dilemma. Public opinion regarding the law which states that the mentally ill, in a psychotic state, are not responsible for their actions, is divided. The study assessed 30 psychiatric patients, committed by court order, following a criminal act on their part. No relationship was found between the nature of their offense and a psychiatric disorder. Patients who committed more serious crimes, such as murder, tended to have committed fewer criminal acts in the past. Sixty-nine percent of the patients think that the mentally ill are not responsible for their actions and 59% agreed with the judge's decision to hospitalize them. On a concrete level, over two-thirds of the patients were able to distinguish right from wrong. The treating physicians related mainly to the patients' illnesses rather than to the crimes for which they were committed. PMID- 9212607 TI - Involuntary hospitalization of delirium patients in Israel: a psychiatric case register study. AB - This paper compares the national Psychiatric Case Register (PCR) data of two groups of delirium patients who were admitted to psychiatric hospitals. One group consists of patients who underwent involuntary civil commitment following a hospitalization order by a district psychiatrist. The second group consists of delirium patients who were voluntarily admitted. During the period 1984-1993, 805 patients with a diagnosis of delirium were admitted to psychiatric hospitals: 710 (88%) were admitted on a voluntary basis, 88 (10.9% were admitted through civil commitment, 7 (0.8%) were admitted in other ways. The two major groups are further analysed regarding demographic, clinical and administrative variables. No statistically significant differences were found between the groups concerning the clinical and administrative variables studied (type of admission, suicide attempt prior to admission, length of hospitalization, type of discharge). Given the prevailing tendency to treat delirium patients in general hospitals, the small number of those involuntarily admitted and the lack of clinical and administrative differences between the groups, the appropriateness of civil commitment procedures regarding delirium patients is questioned. PMID- 9212608 TI - Medical ethics and the executing process in the United States of America. AB - The article focuses on the ethical and moral issues raised by the participation of physicians in the execution process in the United States of America. Discussion centres on two main areas. Firstly, participation in the actual execution, particularly where the method is lethal injection; and secondly psychiatric assessment and treatment of inmates who are deemed not competent in law for execution. It is argued that as an execution is a harm, participation runs counter to the ethics of the medical participation and cannot be justified even on the basis of relieving pain. Treatment of incompetency is permissible in very limited circumstances. Although the assessment of incompetency is not theoretically ethical, practical difficulties may mean participation is justifiable. The issues are discussed in the light of various moral theories including utilitarian and retributivist punishment theories, and the idea of "a right to punishment." PMID- 9212609 TI - Reproductive health law: where next, after Cairo and Beijing? PMID- 9212610 TI - Death and dying in Australia--some medico-legal problems for legislators. PMID- 9212611 TI - Research ethics and HIV/AIDS. PMID- 9212612 TI - Determining the standard of care in medical negligence litigation in Australia. AB - This article reviews the manner in which a court determines whether a medical practitioner has acted in accordance with the appropriate standard of care and considers, in particular, the reliability of the expert evidence which the court relies upon in this regard. The article suggests some options for reform of the way in which expert evidence is being presented to courts. PMID- 9212613 TI - Ethical issues in the planning and conduct of clinical trials of anti-epileptic drugs. PMID- 9212614 TI - Informed consent--should Bolam be rejected? AB - Informed consent requires the person to correctly understand the nature of what is offered and to be free to choose without coercion. Where there are impediments to such decision-making, then appropriate guardians or mental health laws have been provided to enhance protection. Where consent was not fully informed, an injured patient may resort to the tort of battery or negligence for remedy unless the intrusion was as a consequence of emergency treatment to an unconscious patient in the absence of next of kin. This paper reviews the UK Bolam Principle where professional standards were set by peer standards of professional conduct and the US prudent person test in which needs of a prudent patient assume priority. It concludes that there is a need to balance both the rights of patients and obligation of doctors to ensure that justice prevails. It recognises that the final standard of duty of care remains under scrutiny, and that there was a ground swell against absolute adherence to the Bolam Principle, and a need to review the circumstances of each case. PMID- 9212615 TI - Epilepsy: standards of medical care. AB - This paper acknowledges that the majority of doctors who care for those with epilepsy are not specialists in epileptology. It then provides standard guidelines to patient management and provides a flow chart for diagnosis, examination, investigations and treatment to assist in decision making. The paper argues that excessively rigid formalized programmes result in loss of flexibility to respond to changing circumstances, which may require further investigation to confirm the diagnosis and the appropriateness of selected therapy. The paper reaffirms that guidelines could cause unnecessary inhibitions to optimal delivery of care but those who deviate from standard practice must be able to substantiate and justify the approach adopted. PMID- 9212616 TI - Shared care-responsibilities of the doctors. AB - This paper examines 'duty of care' within the context of the sharing of medical management between the family doctor and the specialist consultant. It reviews the referral system which dictates that the consultant has a direct duty of care to the patient while at the same time, needs to meet the obligations of indirect duty of care to maintain proper communications between consultant and fellow-care givers. It concludes that the proximal responsibility dictates that optimal care must be afforded the patient, with full disclosure, informed consent and adequate risk management. Concurrently, there has to be adequate and reliable communication to provide proper supervisory medical care to complement that offered by the specialist. PMID- 9212617 TI - Epilepsy and the law-medical records. AB - This paper reviews the need to keep medical records and concludes that Section 126 of the New South Wales Medical Practice Act, 1992, requires such provision to comply with adequate "professional conduct". This was above and beyond other possible mandatory maintenance of appropriate records, such as may be covered by the notifiable diseases provisions of the Public Health Act. Ethical codes of conduct imposed further obligations to maintain appropriate records, and legal defence against claims of misconduct or negligence required documented evidence to refute false accusations. The emphasis of records has changed with greater need to stipulate risk exposure associated with proposed treatments and advice provided for such things as necessary follow-up. It was further shown that appointment diaries, extra file entries and indications of any failed attendance and resultant subsequent actions were all part of adequate record-keeping. Finally, the paper reviews ownership of medical records and refers to the New South Wales case of Ms Breen, in which it was found that ownership of records, as at the printing of this paper, resided with the doctor. PMID- 9212618 TI - Epilepsy and driving in South Australia--an assessment of compulsory notification. AB - We examined the licensing of drivers with conditions likely to endanger the public in a State in which doctors are obliged to notify the licensing authority. During 1991, 1460 medically endorsed licenses were cancelled. A sample of 245, including all 49 with epilepsy were mostly voluntary, the twenty exceptions with retained files being drivers with epilepsy. In the same period, 115 traffic accidents were attributed to illness, with the only four (4) investigated by the licensing authority being those with licenses endorsed "epilepsy" where a seizure was responsible. Compulsory notification appeared to result in the identification of epilepsy as the important medical reason for controlling licenses, but failed to recognise sleep disorders or alcoholism, both more significant causes of traffic accidents. In those accidents attributed to illness, almost no action was taken to review the medical fitness of drivers, suggesting a reliance on doctors rather than police or road safety authorities. PMID- 9212619 TI - Medical conditions & driving: legal requirements & approach of neurologists. AB - Licensing of drivers with health problems, particularly epilepsy, has medical, social and legal implications that vary from country to country. Legislation and medical guidelines are based as much on empirical as on statistical data. A questionnaire regarding neurological disorders and driving was given to all adult neurologists in Canada (n = 494) and an assessment made of opinions of neurologists working under mandatory reporting legislation compared to those in a discretionary reporting environment. Of 289 (59%) neurologists responding, 50% reported patients with seizures to the Department of Motor Vehicles compared to only 4% for stroke/TIA, 26% for dementia and 8% for other neurological disorders (p < .0001). In the five provinces with mandatory reporting laws, seizures were reported most of the time by 84% compared to only 19% in the five provinces with discretionary reporting laws (p < .0001). An overall minority agreed with mandatory reporting (44%) but this percentage differed in the provinces with and without mandatory reporting legislation (63% vs. 37%, p < .0001). Seizure disorders are selectively reported more often than other neurological conditions. There is considerable variability in the attitude and practice of neurologists in regard to reporting of medical conditions. PMID- 9212620 TI - Investigation of vehicle driving ability in two diagnostic groups of epileptic patients with special neuropsychological approach. AB - The driving abilities of two groups of epileptic patients (temporal lobe epileptics: 44 and idiopathic generalized epileptics: 26) and control group of healthy volunteers were compared. A computerized device (MST-CARAT), was used validated by comparing the test performance measures with the results of the practical driving tests. Our results show that the neuropsychological aspects deserve greater attention in temporal lobe epileptic patients in general and in those epileptic patients receiving non-monotherapy (especially on Phenobarbital). The level of driving skill of well-treated idiopathic generalized epileptic patients was similar to that of normal drivers. PMID- 9212621 TI - Epilepsy and driving license regulations in Slovenia. AB - In Slovenia, an independent European state since 1991, with an area of 20,000 km2 and about 2 million inhabitants-the driving license regulations of former Yugoslavia are still valid. These regulations are very restrictive, requiring a seizure-free period of at least two years without any anti-epileptic drug for private driving. For vocational driving the ban is permanent. In practice epileptologists do not abide by the law (at least as far as private drivers are concerned) and they allow driving if clinical conditions are good and despite therapy. PMID- 9212622 TI - Epileptic drivers--a study of 1,089 patients. AB - A longitudinal study of 1,089 epileptic patients followed up by the same specialist between 1965-1991, allowed close observation of the seizures occurring to the patient at the wheel and their consequences and to relate them to detailed epileptological criteria. The results show road accidents caused by epileptic seizures are few and most of them are minor. The repatriation of risks between patients is very uneven. The quality of the neuro-psychic inter-critical state as well as the patients' degree of compliance seem to be more reliable risk indicators than some more traditional criteria like the length of remission between seizures. Although seizures occur more frequently in patients suffering from Complex Partial seizures as opposed to other forms of epileptic seizures, the differences between patients with epilepsy lies mostly in their behaviour and in their own representation of the risks. There is a need for a body of rules and regulations serving as an official framework regulating the driving test. This widely circulated document should take into account the multiplicity of cases, including the small number of patients thought to be dangerous. Its mode of application should allow doctors as well as patients to opt for a realistic attitude based on decision-making criteria involving a thorough knowledge of epilepsy as well as a thorough knowledge of the psychological characteristics of the patient concerned. PMID- 9212623 TI - A multicenter case-reference study on everyday life risks in epilepsy in Europe. Risk in Epilepsy Study (RESt-1 Group). PMID- 9212624 TI - Epilepsy, the law and the media. AB - It is generally accepted that the media exerts a powerful influence over what the general public feels and believes. This study examined references to epilepsy and the law found in general media communications from January 1993 to December 1994 in the UK. These articles were extracted from general audience material mainly newspapers and magazines. Any article which referred to the legal aspects of epilepsy were then selected. These included those relating to legislation e.g. driving, state benefits and those relating to criminal law e.g. convictions. The reports were analyzed in terms of type of communication, space allotted, circulation and terminology used both in the title and the general text. An attempt was also made to classify the general image which the article portrayed. It is accepted that content analysis itself provides no direct data about the nature of the communicator, the audience or its effects and this method has cautiously been used to classify and describe the manifest content of the method of communication. The study concludes that although it is encouraging that legal aspects of epilepsy are receiving media attention, this attention could be used in a more positive manner in promoting awareness, rather than many of the cases examined which appeared to use negative images in order to pander to popular audience appeal. PMID- 9212625 TI - Regulations prohibiting blood donation by individuals with seizures or epilepsy are not necessary. AB - Throughout the world people who have epilepsy and seizures are prohibited from donating blood. These restrictions are based on the assumption that they are prone to adverse donor reactions, specifically, syncope and convulsions. We describe a study evaluating whether that concern is warranted. During a two year period beginning in 1987, blood donors with a history of seizures were actively recruited by the American Red Cross in the state of Maryland, USA. According to accepted standards, adverse reactions were classified as "slight", for dizziness and nausea without loss of consciousness; "moderate", denoting syncope; and "severe", indicating convulsive syncope. We reviewed a total of 329,143 satisfactory blood donations, and 613 individuals reporting a history of seizures donated blood 723 times. Among donors with seizures, 186 (25.7%) were taking antiepileptic medication, and 61 (8.4%) had one or more seizures in the preceding year. Individuals with seizures had a low incidence of adverse reactions (3.34%). Although slightly higher than the entire population (2.24%), this difference was not statistically significant. In particular, the risk of syncope with or without convulsive activity was low for people with seizures (.21%) and not significantly increased as compared to other donors (.28%). Our study supports the view that individuals with seizures or epilepsy are not at greater risk for adverse reactions after blood donation. Major restrictions on individuals with epilepsy and seizures as blood donors are not warranted. PMID- 9212626 TI - Epilepsy, automatism and the English law. PMID- 9212628 TI - Epilepsy: legal discrimination from negative to positive. AB - Indian law equates epilepsy with temporary insanity and also prohibits a legally valid marriage for a person with epilepsy with inherent risk of divorce. This absurd law, unique to India and possibly Brazil, must be excised in toto. Repeated petitions, by the Indian Epilepsy Association, to the Federal Government, have resulted in only vague assurances and alternate methods are under consideration. There are no legal impediments to education or work. Strict regulations against driving have yielded place to lax rules wherein a person can drive a vehicle, even after a recent fit, provided he gets a certificate from any registered medical practitioner. The nascent medical insurance specifically excludes epilepsy from its ambit. The cost of anti-epileptic drugs includes a 40% tax akin to Value Added Tax in the West. We must consider the impact of these legal impediments on the social fabric of the individual in his/her milieu and vis-a-vis priorities in national development. PMID- 9212627 TI - Automatisms in non common law countries. AB - The distinction made in the common law tradition between sane and insane automatisms, and in particular the labelling of epileptic automatisms as insane, are legal concepts which surprise and even astonish lawyers of other traditions, whether they work within a civil law system or one with elements both from civil law and common law. It could be useful to those lawyers, doctors and patients struggling for a change in the common law countries to receive comparative material from other countries. Thus, the way automatisms are dealt with in non common law countries will be discussed with an emphasis on the Norwegian criminal law system. In Norway no distinction is made between sane and insane automatisms and the plea Not Guilty by virtue of epileptic automatism is both available and valid assuming certain conditions are met. No. 44 of the Penal Code states that acts committed while the perpetrator is unconscious are not punishable. Automatisms are regarded as "relative unconsciousness", and thus included under No. 44. Exceptions may be made if the automatism is a result of self-inflicted intoxication following the consumption of alcohol or (illegal) drugs. Also, the role and relevance of experts as well as the law of some other European countries will be briefly discussed. PMID- 9212629 TI - Epilepsy and employment: the Americans with Disabilities Act and its protections against employment discrimination. AB - People with epilepsy often encounter discrimination. Some cultures have established prohibitive laws limiting access to employment, restricting marriage, and denying driving privileges. It is not enough to repeal such laws. Legislation, such as the Americans with Disabilities Act (ADA), is needed to provide protections against discriminatory practices. The definitions and standards of the law provide clear guidance to employers and workers with disabilities regarding their rights and responsibilities in the workplace. The ADA is proving to be an effective means of addressing allegations of employment discrimination brought forward by people with seizure disorders, although it has not increased the number of people with disabilities who are employed. However, in combination with rehabilitation counseling and employer education, legislative protection can assist people with epilepsy to be more successful in their pursuit of employment opportunities. PMID- 9212630 TI - The case against having "professional privilege" in the physician/patient relationship. PMID- 9212631 TI - Maintenance of professional privilege as exits in France. PMID- 9212632 TI - Epilepsy should not be an accepted defence in criminal proceedings. PMID- 9212633 TI - Epilepsy its place as a legal defence. PMID- 9212634 TI - Sudden death of adults in Japan. AB - Epidemiological features, risk factors and preventive methods of sudden death (SD) derived from studies the authors have performed since 1987 together with colleagues in Niigata University School of Medicine were reviewed. When SD was defined as death occurring within 24 hours of the onset of symptoms, the annual incidence was 145/100,000 for people aged 15 years and older in Niigata Prefecture. The incidence increased sharply along with the advance of age, while the proportion of SD to natural death due to circulatory diseases was higher in younger people. Though diseases of the circulatory system made up approximately 90 percent of all causes of death, SD due to ischemic heart disease was less frequent in Japan than in western countries. SD showed various patterns in seasonal and "within-a-day" occurrences according to sex, age and cause of death. The months of the highest SD occurrence differed by occupation and matched the busiest work periods. A decrease in sleeping hours and mental stress experienced during the preceding week were related to the occurrence of both sudden death and non-fatal acute myocardial infarction. People having structural circulatory diseases were shown to be predisposed to SD when stress occurred, because fatal arrhythmia is easily induced by the above factors in such people. Health examinations were shown to have preventive effects, though limited, against SD. Differences in the resuscitated rates in cases where a witness was present and where one was not indicates that educating people about correct resuscitation methods is important to minimizing SD. PMID- 9212636 TI - Herpesvirus genes: molecular basis of viral replication and pathogenicity. AB - Herpesviruses possess large DNA genomes which contain from approximately 80 to 200 genes. These viral genes are divided into two groups based on whether they are essential or nonessential (dispensable) for virus growth in cell culture: the essential gene products include a set of replication proteins which accomplish the viral DNA replication, while the dispensable gene products include those important in influencing pathogenesis. This article briefly reviews the results of studies relating to the functions and roles of the gene products of human herpesviruses, particularly products associated with the herpes simplex virus. PMID- 9212635 TI - Molecular neurosurgery using gene therapy to treat malignant glioma. AB - In the last decade, the prognosis of brain tumor patients has dramatically improved due to recent advances in microsurgical techniques and the development of functioning neuroimaging, computer-assisted neuronavigation, endoscopic surgery, intravascular surgery and radiosurgery. According to a report by the Committee of Brain Tumor Registry of Japan, the ten year survival rate of patients with benign brain tumors (meningioma, neurinoma and pituitary adenoma) is more than 95%. In contrast, patients with glioma (which constitute 33% of primary brain tumor cases) still have a poor prognosis, especially in the case of malignant (anaplastic astrocytoma and glioblastoma). This poor prognosis is related to the fact that malignant glioma cells aggressively infiltrate into normal brain tissues, making total removal of the tumor impossible. The median survival time of glioblastoma patients is less than 2 years, despite multimodality treatment with extensive surgical resection and adjuvant therapies using radiation and immunochemotherapy. In order to overcome this formidable neoplasm, the effectiveness of molecular neurosurgery using gene therapy has been investigated since 1992. In this paper, molecular genetic studies and the current state of gene therapy for malignant brain tumors are described, and the future direction of this fascinating approach is discussed. PMID- 9212637 TI - Disaster-readiness of medical facilities in Aichi Prefecture. AB - One month following the Great Hanshin Earthquake of January 17, 1995, we conducted a survey of 173 hospitals in Aichi Prefecture to pinpoint problems related to their actual disaster-readiness and the medical backup systems in place to deal with such disasters. This study revealed that staff at 50% of the surveyed hospitals could reach the hospital within an hour, but that communication is almost entirely dependent on phone lines, suggesting that cordless/portable/mobile phones, radio systems, Internet, communications satellites and the like should be studied in the days to come for possible use as effective communication alternatives in times of disaster. Whereas 92% of the surveyed hospitals had manuals dealing with fire outbreaks, other areas were less well represented. For example, only 36.9% of surveyed hospitals had manuals for earthquakes, 31.7% had manuals for power outages and 14.2% had manuals to deal with flooding and water disasters. New manuals must be developed incorporating the key points garnered from experience (especially Hanshin) and be ready for use immediately. It is the time for each hospital to seriously rethink the measures it should take to deal with disasters. PMID- 9212638 TI - Macromotion of the femoral component in artificial hip joint. AB - Macromotion of the femoral component was analyzed in seven loose hips associated with six patients utilizing the manual compression and distraction test. The mean vertical and varus movement of these hips were 6.4 mm and 1.2 degrees, respectively. Out of a total 120 cementless total hip arthroplasties, there was no macromotion in the control group of six patients with moderate to severe thigh pain. There was also no relationship between macromotion and the severity of thigh pain. Micromotion or rotational instability, which could not be analyzed by conventional stress radiograms, may contribute substantially to thigh pain. PMID- 9212639 TI - Human blood lactate and ammonia levels after supramaximal uphill and downhill running. AB - The purposes of this study were 1) to confirm whether there is a difference in the levels of blood lactate and ammonia after supramaximal uphill and downhill running for the same short duration and 2) to examine the relationship between peak blood lactate levels and work/lean body mass (LBM), as well as the relationship between peak blood ammonia levels and work/LBM following supramaximal uphill and downhill running. Eight healthy, untrained male subjects performed supramaximal uphill and downhill running on a motor-driven treadmill for about 70 sec. Though there was a significant difference (p < 0.05) in running speed and work/LBM between supramaximal uphill and downhill running, no significant difference was found in exhaustion time or heart rate. Both the peak blood lactate and ammonia concentrations were significantly lower after downhill running than after uphill running (p < 0.05). Although there was no significant relationship between peak blood ammonia levels and work/LBM following either uphill or downhill running, significant linear relationships between the peak blood lactate levels and work/LBM were observed following uphill running (r = 0.74, p < 0.05) and downhill running (r = 0.72, p < 0.05). These results suggest that the differences in the blood lactate and ammonia concentration between supramaximal downhill and uphill running of the same duration may be due to the total recruitable muscle mass during exercise, and that peak blood lactate can be used as an index of anaerobic work capacity for untrained subjects under these running conditions. PMID- 9212640 TI - An examination of the close relationship between lymphatic vessels and nerve fibers containing calcitonin gene-related peptide and substance P in rat skin. AB - The distribution of nerve fibers containing either calcitonin gene-related peptide (CGRP) and substance P (SP) was investigated in rat skin with special reference to their relationship to the lymphatic vessels. These nerve fibers exhibited a similar distribution pattern but the former were more numerous than the latter. In the dermis and subcutaneous layers, thin nerve fibers containing CGRP or SP were in abundance, and were observed running along the blood vessels as well as freely in the tissue. Nerve fibers with these peptides were often located close to lymphatic capillaries, and innervated lymphatic vessels in the subcutaneous layer, reaching smooth muscles of the tunica media. These findings suggest that some CGRP and SP may directly drain into lymphatic vessels when released under noxious stimulation from nerve fibers around the lymphatic vessels. When discharged from nerve fibers in the vicinity of blood vessels, both peptides may also drain into the lymphatic vessels after causing blood vascular dilation and an increase in permeability producing edema. These peptides may then be transported to the draining lymph nodes where they can modulate the functions of the immune system. PMID- 9212641 TI - Giant cell-rich osteosarcoma: a case report. AB - This report discusses a rare case of giant cell-rich osteosarcoma. The patient, a 19-year-old male, was diagnosed with a metadiaphyseal osteolytic lesion when he consulted a local doctor complaining of motion pain without swelling. Radiography revealed a geographic osteolytic lesion, cortical thinning and ballooning without obvious cortical destruction. However, a fine onion skin-like periosteal reaction was observed on the lateral side of the femur. The transitional none was narrow and endosteal scalloping was also noted. Needle biopsied material clearly showed nuclear atypism of the stromal tumor cells with numerous osteoclast-like giant cells. Using a combination of pathological examination, radiography, computed tomography (CT) and magnetic resonance imaging (MRI), a diagnosis of giant cell rich osteosarcoma was reached. After chemotherapy, resection and limb salvage surgery with an autogeneous autoclaved bone graft, a vascularized fibular graft were performed, and the patient has shown excellent limb function without local recurrence or distant metastasis during the past 72 months. PMID- 9212642 TI - Solid variant of an aneurysmal bone cyst (giant cell reparative granuloma) of the 3rd lumbar vertebra. AB - A 9-year-old girl with a solid variant of an aneurysmal bone cyst in the 3rd lumbar vertebra showed a good response to low-dose radiation as the only treatment. The solid variant of aneurysmal bone cyst is thought to be a reactive response to intraosseous hemorrhage and is also called giant cell reparative granuloma or giant cell reaction. These lesions in the jaw and the short tubular bones of the hands and feet frequently recur after surgery. Aneurysmal bone cysts of the spine also show a fairly high recurrence rate after incomplete resection or radiation therapy. However, 7 previous cases of the solid variant of aneurysmal bone cyst in the spine and this case did not show recurrence after a mean follow-up period of 45 months. This difference in behaviour suggests that the solid variant should be recognized before surgery as being distinct from conventional aneurysmal bone cysts, especially in the spine. PMID- 9212643 TI - Strain-relatedness among different populations of the pathogenic yeast Candida albicans analyzed by DNA-based typing methods. AB - A pathogenic yeast Candida albicans is one of the most frequently isolated microorganisms in patients suffering from opportunistic infection. The typing of the isolates is important not only to elucidate the route of infection but to explore the evolution of the pathogenic yeast. This article presents an overview of recent research on the strain typing in different populations of C. albicans using the following DNA-based methods: electrophoretic karyotyping by pulsed field gel electrophoresis (PFGE), restriction fragment length polymorphism (RFLP) of digests of genomic DNA by restriction enzymes (with or without Southern hybridization with a DNA probe) and amplification of random or specific sequences by using polymerase chain reaction (PCR). The results of these methods were compared for strain discrimination. Studies on the genetic diversity of the strains among different individuals in a single population cohort, and among different populations of healthy carriers and immunocompetent and immunocompromised patients, were reviewed. Typing of the strains recurrently isolated through the episodes of diseases such as vaginitis and AIDS revealed the occurrence of strain variation or microevolution. Typing of isolates from nosocomial infections indicated the possible occurrence of horizontal transmission of the disease within a single hospital. In addition, recent studies suggested a possible mechanism that might involve the C. albicans-specific repetitive sequences, RPSs, for chromosome rearrangements leading to strain variation. PMID- 9212644 TI - Surgery for renal hyperparathyroidism--experience of 640 cases. AB - Our experience in the field of surgery for renal hyperparathyroidism, gained from 640 cases, the largest number in the world, was reviewed. Additional comments were made on the relation between renal transplantation and renal hyperparathyroidism. PMID- 9212645 TI - The role of the ret proto-oncogene in human disease. AB - The ret proto-oncogene encodes a receptor tyrosine kinase with a cadherin-like motif in the extracellular domain. Recently, it turned out that ret is the causative gene for the development of multiple endocrine neoplasia (MEN) type 2A and type 2B and Hirschsprung's disease. MEN 2A and MEN 2B mutations represent activating changes of ret whereas Hirschsprung mutations inactivate ret. In addition, another activating change of ret was found in papillary thyroid carcinoma, particularly in those cancers which developed in children from areas contaminated by the Chernobyl accident. This review summarizes the role of ret in the development of human disease. PMID- 9212646 TI - A live birth from intracytoplasmic injection of a testicular spermatozoon. AB - Testicular sperm was retrieved from a man with complete epididymal obstruction, and intracytoplasmic sperm injection was performed on his wife's oocytes. In four mature treated, two fertilized eggs were obtained, and a clinical pregnancy was established with embryo transfers. One healthy girl (2715 g) was delivered by cesarean section at 38 weeks' gestation. Our case shows that the use of testicular sperm can result in a normal live birth. PMID- 9212647 TI - Surgical outcome of microscopic vasectomy reversal: an analysis of 30 cases. AB - The results of 30 consecutive microscopic vasovasostomy procedures performed at a single institution over a five-year period between 1991 and 1995 were reviewed. When the surgical outcomes of patients who were operated on less than five years after their vasectomy were compared with the outcomes of those patients who received a vasovasostomy more than five years after their vasectomy, decreases in technical success rates were observed as measured by appearance of sperm in ejaculate (56% vs. 36%), biologic recovery as measured by mean sperm counts (56 million vs. 35 million) and mean progressive sperm motility (44% vs. 21%), along with a decrease in clinical success, as measured by overall pregnancy rates (50% vs. 7%, p < 0.05). Therefore, a microscopic vasovasostomy within 5 years of a vasectomy is a favorable procedure for vasectomy reversal. PMID- 9212648 TI - Enhanced acetylcholinesterase in chronic subdural hematomas. AB - Hematomas and specimens were studied from 13 surgically-operated chronic subdural hematoma cases. Evacuated hematomas contained elevated potassium ions and slightly increased levels of Acetylcholinesterase (AChE), and the dural specimens including hematoma capsules stained positive for AChE using histochemical methods, especially in the inner membrane of the dura where the Masson Trichrome staining revealed damaged and irregular collagen fibers. This article suggests that the initial dural damage caused by trauma results in minor bleeding, which in turn causes a raised potassium ion concentration in the hemolytic fluid, which may also cause depolarization and elevated AChE in the dura. PMID- 9212649 TI - An unusual clinical course after mole evacuation: a case report. AB - A 27-year-old woman evacuated a hydatidiform mole at 11 weeks of gestation. Her serum human chorionic gonadotropin (hCG) levels declined progressively but reached a plateau of 2-3 mIU/ml thereafter. The patient was treated with two courses of methotrexate, which did not affect her hCG levels. She refused further chemotherapy and, for more than one year, she was managed expectantly until a significant rise in her hCG titer. Fortunately, an unexpected pregnancy and subsequent missed abortion led to a spontaneous regression of her hCG levels. PMID- 9212650 TI - [Evaluation of the relation of job stress and food intake to hyperuricemia]. AB - To evaluate the relation of job stress and food intake to hyperuricemia, a case control study was performed in male subjects who had undergone complete physical examinations. Cases (n = 113) were those with hyperuricemia of over 7.5 mg/dl and controls (n = 113) were those with serum uric acid of less than 7.5 mg/dl. Stepwise regression analysis was performed using sixteen items which were significantly related to hyperuricemia by McNemar's method. Consequently, four items; "negative attitude toward work (Odds ratio 5.22, p < 0.01)", "tendency to become competitive in the job and other areas (Odds ratio 5.54, p < 0.01)" in type A behavior, "high meat consumption (Odds ratio 6.94, p < 0.01)", and "high fat intake (Odds ratio 4.05, p < 0.01)", were significantly related to hyperuricemia. PMID- 9212651 TI - [Smoking behavior, knowledge and attitudes of freshmen students]. AB - A questionnaire on smoking behavior, knowledge of smoking-related diseases and attitudes toward the passive exposure to smoking was administered and results analyzed for differences in (1) region, (2) major area of study in the university, (3) grade and (4) date of survey on smoking behavior, knowledge and attitude of the freshmen students. (1) 294 urban and 217 provincial university students, (2) 138 freshmen at the Department of Pharmacology and 156 freshmen at the Department of Technology, (3) 136 freshmen of Y. University and 158 freshmen in senior high school of Yamaguchi prefecture, and (4) 217 freshmen surveyed in 1990 and 136 freshmen surveyed in 1995 were the subjects. The results were as follows; 1) The percentage who had smoked once ranged from 0 to 3% among the senior high school girls and female students at minor universities. However, the percentage for male students ranged from 26% to 44% in each survey. There were clear gender differences in smoking behavior. 2) The proportion of students who admitted that they had smoked cigarettes was 30.9% for urban students and 38.6% for provincial university students, but was not a significant difference. There were no significant differences between urban and provincial students regarding knowledge of smoking-related diseases or attitudes toward passive smoke. 3) The proportion of students who admitted that they had smoked cigarettes was 44.0% for the Department of Pharmacology and 26.2% for the Department of Technology, a significant difference. There were no significant differences between pharmaceutical and engineering students in knowledge of smoking-related diseases or attitudes toward passive smoke. 4) The proportion of students who admitted that they had smoked cigarettes was 27.4% for university freshmen and 1.6% for senior high school freshmen. There was a significant difference between the two. The percentage who replied that the smoker must be considerate to non-smokers tended to be higher in the senior high school students than the university students. However, there was no significant differences between the two groups of students in knowledge of smoking-related diseases or attitudes toward passive smoke. 5) Although the proportion of students who admitted that they had smoked cigarettes was 38.6% in the 1990 survey and 27.4% in the 1995 survey, this was not a significant difference. The percentage of students in the 1990 survey who indicated an awareness of the relationship between smoking and coronary heart diseases was significantly greater than the percentage of students who indicated a similar awareness in the 1995 survey. These results suggest that the differences in the grade and the department of the university (or the nature of school) must be considered when surveying smoking behavior. It does not appear to be necessary, however, to consider regional differences or the date of survey, if students were surveyed relatively recently, concerning smoking behavior, knowledge of smoking-related diseases and attitudes toward the effect of passive exposure to smoking. PMID- 9212652 TI - [Health care needs of the elderly in long-term care facilities and their suitable placements for care]. AB - In order to obtain information on the elderly cared for in long-term facilities, regarding their needs and the actual services received in these facilities, and to determine which facility could provide the most suitable service, elderly in two areas, urban S area and rural N area, were surveyed. The results were as follows. (1) The proportion of elderly placed in hospitals where the suitable care placements were hospitals was 84.6% in S area, and 66.6% in N area. (2) In elderly cared for in long-term facilities, the proportion of persons where the suitable care placement was in-home care was 11.7% in S area, and 14.1% in N area. (3) Therefore estimates for the elderly that would be for at home increases by 21.1% in S area and 27.2% in N area than the estimated figures used in plan for Elderly-Health Welfare in 2,000 AD. PMID- 9212654 TI - [Required manpower for health care and nursing services for the aged at home public health nurses, visiting nurses, dental hygienists, dietitians, physical therapists, and occupational therapists]. AB - The purpose of this study was to estimate the manpower required for the health care and nursing services for the aged at home. For prefectural health care and welfare planning for the aged, data such as the proportion of the aged who need help, service demand, and required frequency of services were obtained. The means and "mean +/- 2 x standard deviations" were calculated to obtain various parameters. Calculated figures were those which can be obtained with some effort. The results are as follows (middle level estimation (low level estimation-high level estimation)): requirements are 1.9 (0.61-5.7) public health nurses, 2.6 (0.63-14) visiting nurses, 0.20 (0.084-0.42) dental hygienists, 0.35 (0.17-0.66) dietitians, and 0.25 (0.014-1.27) physical and occupational therapists per population 10,000. For the national total, requirements are 23 (7.3-67) thousand public health nurses, 31 (7.5-160) thousand visiting nurses, 2.4 (1.0-5.0) thousand dental hygienists, 3.9 (2.0-7.8) thousand dietitians, and 3.0 (0.17-15) thousand physical and occupational therapists. By population sizes, for example in the municipalities which has 10-30 thousand people, required are 4.2 (1.7-11) public health nurses, 5.3 (1.3-27) visiting nurses, 0.4 (0.2-0.8) dental hygienists, 0.5 (0.3-0.9) dietitians, and 0.5 (0.0-2.5) physical and occupational therapists. Comparison of the present numbers with estimated manpower needs show that, the present number of public health personnel is almost the same as the low level estimation. But the present numbers of other manpower is lower than the low level estimation. Considering other services such as maternal and child health, it seems that the municipalities which has 10+ thousand population should employ full-time dietitians and dental hygienists. For policy making in a municipality, the policies of other municipalities should be considered. Because it is based on means for municipalities, the results of this study should be useful for application by other municipalities. PMID- 9212653 TI - [Estimation of the future numbers of patients with circulatory diseases in Japan based on the results of national patient surveys]. AB - The numbers of patients aged 35 years or more with hypertension, ischemic heart disease, and cerebrovascular disease 15 years from now were estimated. First, the numbers of patients with these diseases in 1984, 1987, 1990, and 1993 were calculated by age and sex using data from the National Patient Surveys conducted by the Ministry of Health and Welfare. Then, population prevalence for calendar years 1996, 1999, 2002, 2005, and 2008 were estimated based on the past data using linear regression models. Finally, the total numbers of patients were calculated from the estimated prevalence multiplied by the estimated population figure of the national government. The numbers of patients with hypertension will decrease in younger age classes but increase in older age classes; the total number of all patients, therefore, will increase. While the number of patients with ischemic heart disease in males is estimated to increase, that in females will level off. The number of cerebrovascular disease patients in each age and sex group will grow larger. PMID- 9212655 TI - [Prevalence of glucose intolerance, diabetes mellitus, and high serum insulin levels in a Japanese urban population]. AB - An epidemiological study was conducted to investigate the prevalence of diabetes mellitus, glucose intolerance and hyperinsulinemia in an urban population. Twelve thousand two hundred people aged 30 to 79 were randomly selected from residents of S-city in Osaka Prefecture and were urged to attend a cardiovascular examination at the National Cardiovascular Center. In 1992 and 1993, among 5,284 people who received the examination, 75 g oral glucose tolerance tests (OGTT) were performed and plasma glucose and serum insulin concentrations were determined for 2,147 subjects, who participated in the morning course of examination and who were fasting. The prevalence of diabetes and hyperinsulinemia (fasting serum insulin level > or = 15 microU/ml) was higher in older than in the younger generation, and was higher in men than in women. The prevalence of diabetes in those aged 40 and over in S-city was 7.3% in men and 5.6% in women, 17.2% of men and 10.7% of women had impaired glucose tolerance, and 7.5% of men and 5.2% of women had hyperinsulinemia. A comparison of prevalences of diabetes was performed between 4 populations, one being our urban population and others being 3 rural populations where population-based surveys had been accomplished with OGTT but without a screening as ours was with regard to detecting diabetes. The prevalence of diabetes in our urban population did not appear to be higher than in the other 3 rural populations. From a questionnaire survey of responders and non-responders to OGTT, it was considered that the degree of selection bias in this study was small, if any. PMID- 9212656 TI - [Reevaluation of heart disease deaths on death certificates and trends for ischemic heart disease mortality during the last five years in Oita city]. AB - To evaluate heart disease deaths and clarify trends for ischemic heart disease (IHD) mortality during the last five years in Oita City, we reevaluated causes of death on death certificates. In 1993, there were 253,000 people aged 25-74 in Oita City. In this population age group, there were 1,996 deaths from January 1992 through December 1993. Our subjects were 982 deaths recorded as caused by heart disease and IHD related diseases. Subjects were reevaluated on the basis of physician's interview, clinical records and police records. This IHD reevaluation was conducted by the WHO MONICA criteria. The death certificates identified 321 heart disease deaths, of which there were 80 (24.9%) acute myocardial infarctions (AMI), 22 (6.9%) other IHD, 180 (56.1%) heart failures, and 39 (12.1%) other heart diseases. The remaining 61 deaths were caused by other diseases. After reevaluation, 40 'definite' AMI and 60 'possible' AMI were recognized through the MONICA criteria, 86 sudden deaths (SD) which were defined as, 'death within 24 hours of the onset of acute symptoms and without clear signs suggesting what disease was the cause,' were also classified. Assuming that 50% of SD were due to IHD, according to some postmortem autopsy studies, aged-standardized IHD mortality per 100,000 for males was 38.3 per year and for females 17.3 per year in this period. Both mortality rates, 31% for males and 38% for females, were higher than IHD mortality statistics. Moreover, in comparison with IHD mortality estimated by reevaluation of heart disease in Oita City in 1987-88, age standardized IHD mortality per 100,000 for males has remained basically stable, increasing from 37.8 to 38.4 during the last five years. On the other hand, mortality for females has increased from 11.2 to 17.3. Our results suggest that mortality from IHD actually was about 30% more than mortality statistics, and does not show a declining trend as mortality statistics have stated. PMID- 9212657 TI - [Use of casual plasma glucose levels in community screening for diabetes mellitus]. AB - The aim of this study was to assess the utility of casual plasma glucose as a screening test for diabetes mellitus in lieu of hemoglobin A1c (HbA1c) and urinary glucose. Casual plasma glucose and urinary glucose were measured in 1,024 individuals in a rural community. The individuals were then offered participation in an oral glucose tolerance test (OGTT). A 75 g OGTT based on WHO criteria was performed and HbA1c was measured in 290 respondents. Receiver operating characteristic (ROC) curves for casual plasma glucose and HbA1c were constructed and the areas under ROC curves for the tests were calculated. The area under the ROC curve for HbA1c was significantly higher than that for casual plasma glucose (p < 0.01). The sensitivity and specificity for casual plasma glucose > 140 mg/d/were 61% and 62%, respectively, while the specificity of HbA1c was 93% with a sensitivity of 61%. The sensitivity of urinary glucose was low (only 18%) with a comparable specificity (93%). It is concluded that measurement of casual plasma glucose is inferior to HbA1c as a screening test for diabetes. PMID- 9212658 TI - [A study on competencies used by public health nurses in creating new health care systems in the community]. AB - The purpose of this study is to clarify the competencies used by public health nurses in creating new health care systems in the community. The subjects were four public health nurses who were nominated by other public health nurses. As a method a qualitative method was used. Each subject was interviewed using a semistructured questionnaire which was made based on each system she had developed. Each interview was tape recorded and transcribed verbatim. Data were analyzed and divided by the seven domains proposed by Benner. The results showed that public health nurses took roles in six out of the seven domains of Benner. Among them the "Organizational and Work-Role Competencies" domain had a lot of content and was divided into more detailed seven subcategories. All four public health nurses were stimulated to create a new care system by exposure to a serious case and being moved deeply through visits to homes and hospital. The desire to solve the problem of individual clients leads to the establishment a new system in the community, showing the originality of the role of public health nurses who have nursing knowledge and who work in a regional government. In the process of building a health care system each public health nurse secured the budget or created a new measure of the regional government. It shows the significance of the role that public health nurses play in the regional government. PMID- 9212659 TI - [Children's welfare system in the U.K.--current multiagencies aspects]. PMID- 9212660 TI - [Length of survival of elderly patients with chronic obstructive pulmonary disease]. AB - We examined survival in 53 patients over 60 years old who had chronic obstructive pulmonary disease, and whose FEV1 was less than 60% of the forced vital capacity and was less than 60% of the predicted value. They comprised 34 men and 19 women. The mean age was 75.2 years. The %FEV1 was 39.0% and the body mass index was 19.0. Neither age %FEV1, nor body mass index differed significantly between men and women. However, the number of cigarettes smoked per day, the number of years of cigarette smoking, and the Brinkman index were higher in men than in women. The Brinkman index was 1255.0 in men and 617.8 in women (p = 0.0001). For the group as a whole, the 5-year survival rate was 65% and the 10-year survival rate was 35%. The survival rate of men did not differ from that of women. Survival and %FEV1 did not differ between men and women, despite the significant difference in Brinkman index, which suggests that women were more susceptible to the effects of cigarette smoking than men. Age, one tenth of %FEV1, and body mass index less than 19 were found to be independent predictors of mortality (proportional hazards analysis, p = 0.044, 0.019, and 0.024, respectively). The 5-year survival rate were as follows: 85% in patients less than 75 years of age, 50% in patients more than 75 years of age, 25% in patients with a %EFV1 less than 30%, 80% in patients with a %FEV1 of 30% to 49%, 62% in patients with a %FEV1 of 50% to 60%, 50% in patients with a body mass index of less than 19, and 83% in patients with a body mass index of more than 19. PMID- 9212661 TI - [Development of core/targetoid fibers in the diaphragm of chronically hypercapnic rats]. AB - We tested the hypothesis that chronic hypercapnia contributes to the development of core/targetoid fibers in ventilatory muscles. Five Wistar rats were kept in 5% CO2 in air for 13 weeks. Arterial blood gases (mean +/- SE) were pH 7.402 +/- 0.017, PaCO2 47.4 +/- 2.4 Torr, and PaO2 105.2 +/- 13.2 Torr. While the rats were chronically hypercapnic, their mean minute ventilation was about 1.7 times higher than that of rats that breathed air. This level was maintained throughout the exposure to CO2. Approximately 6% to 12% of the diaphragmatic type-I fibers were replaced by core/targetoid fibers in 3 of 5 chronically hypercapnic rats but not in the 3 control rats. Chronic hypercapnia may have continuously stimulated the respiratory center and resulted in more work done by the ventilatory muscles. Long-term overuse of the ventilatory muscles, which may occur in patients with chronic respiratory failure, can cause morphological changes in type-I fibers in the diaphragm. PMID- 9212662 TI - [Pneumonitis induced by the herbal medicine Sho-saiko-to in Japan]. AB - We studied the clinical characteristics of pneumonitis induced by Sho-saiko-to (SST). Of 94 cases reported to a drug maker, 72 were judged to be SST-induced pneumonitis (52 men and 20 women, mean age 63.7 years). Most patients took SST for chronic liver diseases due to infection with the hepatitis C virus. The mean duration of SST therapy before the onset of pneumonitis was 50.2 +/- 42.1 days. Most patients presented with coughing, dyspnea, and fever of acute onset. Chest X ray films showed diffuse ground-glass shadows and infiltration. Abnormally high levels of C-reactive protein and lactate dehydrogenase were common, as was hypoxia. Analysis of bronchoalveolar lavage fluid revealed abnormally high percentages of lymphocytes and neutrophils and a low CD4/CD8 ratio. Although 64 of 72 patients survived after cessation of SST only or steroid therapy, 8 died of respiratory failure despite high-dose steroid therapy. Compared with patients who survived those who died were more likely to have an underlying lung disease, had been taking SST longer after the onset of pneumonitis, and had more severe hypoxemia. PMID- 9212663 TI - [Increase in pulmonary vascular permeability caused by increased expression of Mac-1 on the surface of polymorphonuclear leukocytes]. AB - We studied the expression of adhesion molecules on the surface of human polymorphonuclear leukocytes (PMNs). The effects of mechanical stimulation were measured with a flow cytometer and pulmonary vascular injury due to accumulation of PMNs in the lungs was assessed by a gravimetric method. The accumulation of PMNs in the lungs was studied by measuring the amount of myeloperoxidase. PMNs were stimulated by gentle agitation in a glass container for 10 s. Mac-1 (CD11b/CD18) was upregulated on the surface of PMNs that were mechanically stimulated. When unstimulated PMNs were exposed to isolated rat lungs, the filtration coefficient did not change from that under baseline conditions. However, when mechanically stimulated PMNs were exposed to isolated rat lungs, the filtration coefficient was about 5 times higher than that measured at baseline. When mechanically stimulated PMNs treated with anti-CD18 antibody were used, the increase in the filtration coefficient was completely blocked. The assay of myeloperoxidase revealed that PMNs stuck to isolated rat lungs only after stimulated PMNs were added. We conclude that when the adhesiveness of PMNs is increased by mechanical stimulation, these cells adhere to pulmonary vessels and increase pulmonary vascular permeability. PMID- 9212664 TI - [Acute pulmonary thromboembolism treated with E6010]. AB - A 22-year-old man was admitted to our hospital because of sudden dyspnea and dizziness. Hypoxemia was found. Lung perfusion scintigraphy and pulmonary angiography showed massive pulmonary thromboembolism. The patient received E6010, a derivative of tissue plasminogen activator by intravenous injection for about 2 minutes. One hour after this treatment, pulmonary angiography showed lysis of the ciot, the pulmonary arterial pressure had decreased, and the cardiac index and PaO2 had increased. Despite anticoagulant therapy, pulmonary embolism recurred so we implanted a Greenfield filter in the inferior vena cava. This was the first case of pulmonary thromboembolism in which E6010 had a beneficial effect. We were also able to document hemodynamic and radiologic changes after intravenous infusion of this drug. Recurrent pulmonary embolism is an indication for filter placement, and this patient will need a long period of follow-up. PMID- 9212665 TI - [Conversion of chronic necrotizing pulmonary aspergillosis to invasive pulmonary aspergillosis, and successful treatment with fluconazole]. AB - An 82-year-old man with rheumatoid arthritis was admitted to the hospital because of bloody sputum, appetite loss, and a chest-radiographic abnormality. He had previously been treated with oral steroid therapy. The chest X-ray film showed an infiltrative shadow with airlucency in the right upper lung field. Sputum culture for fungi was negative, but a test for aspergillus antigen in serum was positive. Other clinical findings were also compatible with conversion of chronic necrotizing pulmonary aspergillosis to invasive pulmonary aspergillosis. The patient was successfully treated with a drip infusion of fulconazole. The patients condition was stable for several months, after which he died due to uncontrollable atrial flutter. Mild immunosuppression due to oral steroid therapy probably caused chronic necrotizing pulmonary aspergillosis in this case. The patient's general condition worsened after admission and invasive pulmonary aspergillosis developed. This case taught us that therapy for chronic necrotizing pulmonary aspergillosis should include management of the patient's general condition as well as treatment of the pulmonary lesions. PMID- 9212666 TI - [Tumor lysis syndrome after treatment for mediastinal non-Hodgkin's lymphoma]. AB - An 18-year man was admitted to the hospital because of acute dyspnea. Roentgenological examination revealed a large anterior mediastinal tumor. Histologic examination of a specimen from a cervical lymph node yielded a diagnosis of non-Hodgkin's lymphoma of, diffuse, large, T-cell type. Acute respiratory failure and a massive pleural effusion developed, and mechanical ventilation was begun. Chemotherapy with adriamycin, vincristine, cyclophosphamide, and prednisolone resulted in rapid shrinking of the mass. Acute renal failure developed because of hypoperfusion of the kidney caused by acute circulatory failure and the tumor lysis syndrome, and rapid increases in the concentrations of lactate dehydrogenase, creatine phosphokinase, and uric acid in serum after the tumor collapsed. Mediastinal malignant lymphoma often forms a bulky mass, and effective chemotherapy, while it can prolong survival, may also cause the tumor lysis syndrome. PMID- 9212667 TI - [Suspected leiomyosarcoma of the trachea]. AB - A 73-year-old women was admitted to our hospital because of dyspnea, bloody sputum, and an apparent intratracheal tumor. A chest X-ray film from August 1991 showed a sharply circumscribed soft-tissue density in the trachea at the level of the aortic arch. By March 1993 the tumor had grown from 15 mm to 20 mm in diameter. A chest CT scan and fiberoptic bronchoscopy showed an intratracheal tumor that occupied 80% to 90% of the tracheal lumen. The tumor was resected on March 26, 1993. Histopathological study revealed a suspected leiomyosarcoma of the trachea. Only 18 cases of leiomyosarcoma of the trachea have previously been reported worldwide. We know of only 3 previous reports of the case of suspected leiomyosarcoma of the trachea in Japan. PMID- 9212668 TI - [Diffuse bronchiolo-alveolar cell carcinoma that produced both amylase and CA19 9]. AB - A 68-year-old man who worked as an editor was admitted to Aichi Medical University Hospital due to dyspnea on exertion and emaciation. The patient had noticed rapid weight loss during diet therapy for diabetes mellitus that started in the beginning of July, 1993. Laboratory examinations revealed elevated levels of LDH and amylase in serum. Ultrasonography disclosed minimal ascites. Dyspnea on exertion developed in September, 1993. Chest roentgenography showed diffuse bilateral small nodular or reticular opacities. CT-guided percutaneous needle aspiration was done and cytologic examination of a specimen of lung tissue revealed papillary adenocarcinoma. The diagnosis was bronchiolo-alveolar carcinoma. Serum levels of amylase were elevated. The amylase isozyme pattern was of the salivary type. Serum levels of CA19-9 and CEA were also elevated. The patient died of respiratory failure on December 4, 1993. Postmortem examination revealed diffuse small nodules in both lungs. Examination of the nodules showed bronchiolo-alveolar cell carcinoma. The tumor cells stained positively for amylase (salivary type, not pancreatic type) CA19-9, and CEA by the avidin biotin complex method, but they were immunohistologically negative for AFP. We conclude that this lung cancer produced amylase, CA19-9, and CEA. We know of only a few reports of cases in which lung cancer produced both amylase and CA19-9. PMID- 9212670 TI - [Autopsy findings of retroperitoneal cystic tumor and peliosis hepatis in lymphangiomyomatosis]. AB - A 40-year-old woman who worked as a nurse and had suffered from progressive exertional dyspnea for about 14 years underwent open lung biopsy with surgical treatment for pneumothorax. The diagnosis was lymphangiomyomatosis and she was treated with danazol to suppress ovarian function. Her condition improved temporarily, but she died of respiratory failure when she was 47 years old. The survival time after the onset of respiratory symptoms was 21 years, and after the biopsy it was 8 years. At autopsy a retroperitoneal cystic tumor was found (9 x 7 x 5 cm), which had been evident clinically. Histologic examination showed that the tumor was an extrapulmonary manifestation of the lumphangiomyomatosis lesion. Some paraaortic lymph nodes has similar lesions. Aggregates of small red spots were seen on acute surface of the liver. These were diagnosed as peliosis hepatis, they may have been caused by the danazol. PMID- 9212669 TI - [Peripheral carcinoid tumor of the lung presenting as a solitary mass shadow on a chest roentgenogram and diagnosed by CT-guided percutaneous needle biopsy]. AB - A 59-year-old woman was referred to our clinic because a solitary mass shadow (2.5 cm in diameter was seen in the left lower lung field on a chest roentgenogram. Transbronchial biopsy via a fiberoptic bronchoscope did not yield a definitive diagnosis. Examination of a specimen obtained by CT-guided percutaneous needle biopsy revealed that the tumor consisted of small cells proliferating in a solid, tubular, pseudoglandular, and follicular pattern, which suggested the diagnosis of carcinoid. A left lower lobectomy was performed, and several hilar and mediastinal lymph nodes were also removed. The tumor cells did not show pleomorphism of nuclei, mitotic activity, or necrosis. Grimelius and immunohistochemical stains for chromogranin. A, neuron-specific-enorase, synaptophysin, and serotonin were positive. These findings confirmed the diagnosis of carcinoid CT-guided percutaneous needle biopsy can be useful for the diagnosis of peripheral carcinoid and other peripheral lung tumors. PMID- 9212671 TI - [Removal of bronchial foreign bodies by suction with a bronchoscope]. AB - We report two cases in which intrabronchial foreign bodies were removed with a fiberoptic bronchoscope. In both cases the foreign body was a seed of a small Japanese apricot. Atelectasis or obstructive pneumonia was seen on chest roentgenograms. The foreign bodies were associated with slight inflammation and polyps on the bronchial epithelium. The foreign bodies were removed by applying suction with a fiberoptic bronchoscope. This method may also be useful for removing other large, hard, uneven, and ball-like foreign bodies. PMID- 9212672 TI - [Bilateral hilar and mediastinal lymphadenopathy accomporying pulmonary infiltration with eosinophilia]. AB - A 46-year-old man had had an occasional dry cough in the early morning since about the age of 20, but had received no treatment. He had been taking an antirheumatic drug for 2 years for rheumatoid arthritis. The patient complained of fever and dry coughing that began in the middle of November 1995, and he was treated for acute bronchitis. His condition did not improve, and he was admitted to the hospital in early December. Wheezing and rhonchi were heard in both lung fields. His white blood cell count was 19,000/mm3, and the eosinophil percent age was 48%. A chest CT scan revealed macular lesions with an increased density in both lung fields, and markedly swollen mediastinal and hilar lymph nodes. Analysis of alveolar lavage fluid revealed an increased number of cells (total) and eosinophilia (37%), and examination of a transbronchial lung biopsy specimen indicated infiltration with eosinophils and lymphocytes. Our diagnosis was eosinophilic pneumonia. The patient's condition improved soon after the start of pulse therapy with steroids. Bilateral swelling of mediastinal and hilar lymph nodes is rare in patients who have pulmonary in filtration with eosinophilia (the PIE syndrome). PMID- 9212673 TI - [Large-cell carcinoma presenting as diffuse thickening of the pleura and resembling malignant mesothelioma]. AB - A 65-year-old man had a right-sided pleural effusion. A thoracic CT scan revealed diffuse pleural thickening resembling malignant mesothelioma. Repeated needle biopsies did not yield a definitive diagnosis. At autopsy, diffuse pleural thickening was found, and examination of a specimen of lung tissue showed large cell carcinoma. Hyalinized fibrous tissue with malignant cell infiltration was seen in the thickened pleural tissue and metastatic pleuritis was diagnosed. Large-cell carcinoma of the lung should be considered in the differential diagnosis of pleural thickening. PMID- 9212674 TI - [Pulmonary infiltration with eosinophilia due to rabbit-fur antigen: diagnosis by allergen inhalation test]. AB - We report the case of a 48-year-old man with asthma and pulmonary eosinophilia. He presented with coughing, dyspnea, and wheezing that began 6 months after he began keeping a rabbit in his house. He was referred to our department for further examination of pulmonary infiltrative shadows. An inhalation test with rabbit-fur antigen induced both immediate and late asthmatic responses. In addition, infiltrative shadows appeared in the right segments 2, 8, and 9 on chest CT films after the antigen inhalation. Examination of fluid obtained by bronchoalveolar lavage from the right S9 showed an increase in the fraction of eosinophils. Examination of a specimen obtained by transbronchial lung biopsy from those segments revealed infiltration of inflammatory cells into the alveolar septa and alveolar spaces, which was consistent with eosinophilic pneumonia. Our diagnosis was asthma and pulmonary eosinophilia due to rabbit-fur antigen. PMID- 9212675 TI - [Bronchopneumonia caused by Neisseria meningitidis--probable transmission by a family member who had been in Hong Kong]. AB - A 22-year-old man came to our hospital complaining of bloody sputum. No abnormalities were seen on a chest X-ray film, but computed tomography of the chest showed infiltrates in the middle and lower lung fields. Neisseria meningitidis was isolated from cultured samples of naso pharyngeal secretions, from sputum, and from a sample obtained during bronchoscopy; bacterial bronchopneumonia was diagnosed. Family members were studied for meningococcal infection, and the bacterium was cultured from a nasopharyngeal smear from the patient's mother. The mother was asymptomatic. Both bacteria belonged to Group B serum group and their drug sensitivities were equal. The patient's family had been in Hong Kong, where this bacterium is endemic, for two years. They then returned to Japan and started to live together with the patient nine months before the onset of symptoms. The mother may have become infected in Hong Kong and may have passed the bacterium to her son after her return. Oral ampicillin was effective in both mother and son. Investigation of the patient's family was necessary to establish the probable route of infection and to decide on the treatment. PMID- 9212676 TI - [Lung cancer with a sarcoid-like reaction in the primary tumor]. AB - An 80-year-old woman was admitted to the hospital because of a nodular lesion in the right upper lobe of the lung. Transbronchial biopsy was performed and adenocarcinoma of the lung was confirmed by pathological examination. The tumor was resected by right upper lobectomy and was found to be a moderately differentiated tubular adenocarcinoma. Numerous non-caseating epithelioid cell granulomas were also found intermingled with the cancer cells. Metastasis was apparent in several regional lymph nodes but no granulomatous lesions were found in any lymph node, regardless of metastasis. These findings were compatible with a "sarcoid-like reaction" because there was no clinical evidence of generalized sarcoidosis or pulmonary mycobacterial infection. Although sarcoid-like reactions are occasionally associated with cancer, formation of an epithelioid cell granuloma inside the primary tumor is very rare. All the reported cases of a sarcoid-like reaction within the primary lung tumor so far were with adenocarcinoma. The sarcoid-like reaction may be a local immune response to the cancer cells. PMID- 9212677 TI - [Use of a bronchoscope for thoracoscopic observation and diagnosis of pleural plaques]. AB - A 62-year-old man was admitted to our hospital for evaluation of bilateral tumor like shadows along the ribs. He had been a diamond-grinder for twenty years. Thoracoscopic examination with a flexible bronchoscope revealed many well circumscribed tumors with shiny, smooth, convex surfaces on the parietal pleura. The biopsy specimen had hyaline collagen with no cellular components. Based on these findings the tumors were diagnosed as pleural plaques. Cases of pleural plaques are usually followed up with chest radiography only, but the clinical course may be complicated by malignant mesothelioma and other malignant disease. To differentiate these conditions we performed biopsy during intrathoracic observation. The flexible bronchoscope can be useful in such procedures because it is relatively easy to operate and the procedure is relatively non invasive. PMID- 9212678 TI - [Tracheobronchopathia osteoplastica with "rock garden" findings on bronchoscopy]. AB - We encountered a 65-year-old man with tracheobronchopathia osteoplastica with "rock garden" findings on fiberoptic bronchoscopy. Many nodular elevated lesions were seen on all sides of the trachea and main bronchi except the membranous portion. The diagnosis was confirmed by examination of a biopsy specimen. PMID- 9212679 TI - [Effectiveness of prolonged oral therapy with low-dose etoposide in patients aged 85 years and over with non-Hodgkin's lymphoma]. AB - Individualizing the doses of cancer chemotherapy agents and progress in supportive therapy have improved the prognosis for elderly patients with non Hodgkin's lymphoma. Prolonged hospitalization elderly patients has adverse effects, which include dementia, difficulty in walking, and depression. We treated 10 elderly patients (> or = 85 years) with non-Hodgkin's lymphoma as outpatients with oral etoposide, 25 mg or 50 mg daily for as long as possible, or until the white blood cell count decreased to < or = 2,000/microliter or the platelet count decreased to < or = 5 x 10(4)/microliter. Complete remission was achieved in 4 patients and partial remission in 4; the median duration of survival was 19 months. Adverse effects included leukopenia in 1 patient (< or = 1,000 cells/microliter), thrombocytopenia in 1 patient (< or = 5 x 10(4) cells/microliter), and anorexia in 1 patient. These results indicate that prolonged oral administration of low-dose etoposide is effective and safe for the treatment of non-Hodgkin's lymphoma in elderly patients. This outpatient chemotherapy caused no serious adverse reactions. PMID- 9212680 TI - [Tokyo Centenarian Study. 4. Apolipoprotein E phenotype in Japanese centenarians living in the Tokyo Metropolitan area]. AB - To examine the relationship between apolipoprotein E (apoE) phenotype and life span, we measured the frequently of the apoE phenotype and allele in 54 Japanese centenarians who lived in the Tokyo metropolitan area in 1994, 1995, and 1996. The control group consisted of 973 subjects, 883 healthy volunteers who were described previously and 90 healthy people who came to the Keio health consulting center. The apoE phenotypes in the centenarians was 2 E2/E2 (3.7%), 5 E2/E3 (9.3%), 38 E3/E3 (70.4%), and 9 3E/E4 (16.7%). No other phenotype was observed. In the control group, the phenotypes were 2 E2/E2 (0.2%), 57 E2/E3 (5.9%) 712 E3/E3 (73.2%), and 179 E3/E4 (18.4%). The frequency of E2 was higher in the centenarians. The frequencies of the apoE allele in the centenarians and the control subjects were epsilon 2 8.3% vs. 3.5%, epsilon 3 83.3% vs. 85.4%, and epsilon 4 8.3% vs. 10.9%. The frequency of the apoE allele differed significantly between centenarians and control subjects (chi 2 = 6.84, p = 0.033). Levels of serum cholesterol and apolipoprotein B were significantly lower in the E2/E2 + E2/E3 centenarians. Studies of the frequency of the apoE allele in Japanese, French, and Finnish subjects showed that epsilon 2 is more frequent and epsilon 4 is less frequent in centenarians. These data show the apoE phenotype may affect life span: epsilon 2 is positively and epsilon 4 is negatively associated with longevity. PMID- 9212681 TI - [Prevention of cardiac events by beta-blocking agents in elderly patients after myocardial infarction]. AB - We retrospectively analyzed the effect of beta-blocking agents on the incidence of cardiac events in elderly patients who had had myocardial infarction. A total of 1169 patients who had had a myocardial infarction (age, 60.2 +/- 11.4 years) were followed for a mean of 18.0 +/- 19.7 months and the incidence of cardiac events (fatal or nonfatal myocardial infarction, sudden cardiac death, and death due to congestive heart failure) was computed. There were 21 cardiac events in 653 patients who received beta-blocking agents (3.2%) and 39 events in 516 patients who did not receive beta-blocking agents (7.6%, p < 0.01). Among patients less than 50 years old, the incidences of cardiac events were 4.1% in those who received beta-blocking agents and 7.6% in those who did not; among those 50 to 59 years old the incidences were 3.0% and 7.5%, respectively; among those 60 to 69 years old they were 4.3% and 6.0%, respectively; and among those 70 years old or older they were 0.8% and 11.4%, respectively (p < 0.01). We found that beta-blocking agents prevented cardiac events both in elderly and in younger patients after myocardial infarction. PMID- 9212682 TI - [Development of comprehensive assessment system for elderly patients in a geriatric hospital]. AB - Studies in the United States and Europe revealed that a comprehensive geriatric assessment is useful in the management and treatment of elderly disabled patients. In Japan, there are few reports of the development of such assessment. We examined the reliability of a scale designed to assess the performance of activities of daily living (ADL), a revised version of Hasegawa's Dementia Scale) HDSR, and a depression scale (GDS-15), using results from 140 patients over 65 years old who were admitted to our hospital from the end of January to early February 1994. The reliabilities of the ADL and HDSR scales were acceptable, but the kappa value of the GDS-15 was very low. The ADL score was low in patients with cerebrovascular diseases, dementia, and bone and joint problems, but the HDSR score was not low in the latter group. These results suggest that each disease has a characteristic pattern of impairments and disabilities. Although psychological assessment is essential and requires validation, a prospective study of outcomes in the evaluated elderly patients might also be fruitful. PMID- 9212683 TI - [Relation of asymptomatic myocardial ischemia to impaired communication, cerebrovascular disease, and lack of ability to perform activities of daily living in elderly patients]. AB - Patient's reports of angina pectoris depend on cognition and communication, and on the patient's physical and mental activity. In elderly people, these functions are often impaired, and we therefore looked for evidence of myocardial ischemia in 770 consecutive autopsies. We defined the coronary stenosis index (CSI) as the sum of the stenosis scores of three coronary arteries: 0% = 1, 25% = 2, 50% = 3, 75% = 4, 90% = 4.5 95 or 100% = 5. A total score of more than 13.0 and a score of 5.0 in one vessel was assumed to have been associated with myocardial ischemia. Patients assumed to have had myocardial ischemia were classified according to the presence or absence of angina pectoris: 24 had angina pectoris and 92 were asymptomatic. As controls, 86 patients in whom the CSI was lower than 10.0 were studied. Death due to myocardial infarction was most frequent in patients with angina (67%). Acute myocardial infarction was more common in asymptomatic patients than in controls (27% vs. 1%). Small myocardial infarctions and inferior myocardial infarctions were more frequent in asymptomatic patients than in those with angina. Cerebrovascular disease, problems doing activities of daily living, and communication disturbance were more common in asymptomatic patients than in those with angina. Electrocardiographic evidence of an old myocardial infarction was found in 40.9%, 16.6%, and 2.3% of patients with angina, asymptomatic patients, and controls, respectively (p < 0.05). Morphologic details of the myocardial infarction, coexistence of cerebrovascular disease, inability to perform activities of daily living, and impaired communication are associated with asymptomatic myocardial ischemia. Patients with asymptomatic myocardial ischemia should be identified and treated because of their relatively poor prognosis. Ischemic events might be detected by careful observation and prevented by appropriate treatment. PMID- 9212684 TI - [Geriatrics in the new program for initial postgraduate medical training in Japan]. AB - Until 1993, initial postgraduate medical training in Japan was conducted only at university hospitals or other teaching hospitals. In 1994, the number of types of training sites was expanded to include health centers, nursing homes, and extended care facilities, which gave trainees chances to understand primary medical care in various settings, providing in-home as well as other non-hospital medical services. Since the new program seemed to increase training opportunities in geriatric medicine, questionnaires regarding the new program were sent by mail to all teaching hospitals in Japan (261); 213 responded. The results showed that 29 hospitals already had a program of care for the aged. The teaching sections were mainly internal medicine, psychiatry and rehabilitation medicine. Fifty hospital had plans to include care for the aged in their program in the near future. The number of programs that includes training in care for the aged is gradually increasing. It is hoped that during their initial postgraduate training many young doctors will come to recognize the importance of geriatric medicine. PMID- 9212685 TI - [Effects of administration of Clostridium butyricum to patients receiving long term tube feeding]. AB - In patients who require total parenteral or enteral nutrition the intestinal lining may atrophy and the ability to absorb nutrients may be lost. To prevent atrophy of the small intestine, we administered a suspension of Clostridium butyricum to elderly patients receiving tube feeding, and then measured the activation of serum diamine oxidase and the number, form, water content, and bacteria content of stools, indicators of intestinal structure. We found a significant increase in diamine oxidase activity and an improvement in stool condition: the number of stools per day decreased, form improved, and water content and the number of aerobic bacteria decreased significantly. These results indicate that in patients receiving long-term tube feeding, administration of Clostridium butyricum can restore condition to a near-normal state. PMID- 9212686 TI - [High levels of serum ferritin in elderly patients with non-insulin-dependent diabetes mellitus]. AB - We studied concentrations of serum ferritin, glycosylated ferritin, and non glycosylated ferritin in elderly patients with diabetes. The subjects were 111 people who were at least 60 years old: 54 healthy controls, 14 diabetic patients without retinopathy, and 43 diabetic patients with retinopathy. The mean levels of ferritin, glycosylated ferritin, and non-glycosylated ferritin in serum were significantly higher in the patients with retinopathy than in healthy controls. The mean percent glycosylated ferritin did not differ between patients with retinopathy and healthy controls. The mean levels of serum ferritin, glycosylated ferritin, and non-glycosylated ferritin, and the percent glycosylated ferritin did not differ significantly between patients without retinopathy and health controls. None of these values differed between subjects with macroangiopathy and those without macroangiopathy, in both groups of patients. In patients with diabetes, none of the values measured was significantly related to fasting plasma glucose, HbA1c, or the duration of diabetes. These results suggest that diabetic microangiopathy is associated with abnormally high levels of ferritin in serum. PMID- 9212687 TI - [Longitudinal and comprehensive follow-up study of the oldest man in Japan]. AB - The oldest man in Japan reached the age of 112 years in October 1996. As an Okinawan centenarians, he had been followed closely for the previous 12 years. One sister, 8 years younger, was alive at the start of the study; all other family members were killed in the Okinawan War, 1945. The man did agricultural work until age 85, after which he continued to be physically active and to pay close attention to his health. Results of medical examinations, including blood tests, remained within the normal limits, with a few exceptions. Some abnormalities were found on the electrocardiogram; the red blood cell count and the hemoglobin and hematocrit values decreased relatively slowly. His intake of nutrients was relatively well-balanced, and at the age of 100 his intake energy was 1361 kilocalories per day, which is close to the value recommended for centenarians. His personality was categorized as "Type A", but the pattern was typical of that seen in other Okinawan centenarians. He was able to perform almost activities of daily living until the age of 108. At that time he was admitted to the hospital and his ability to perform those activities decreased sharply. His scores on the revised version of the Hasegawa dementia scale was within the normal range when he was 106 years old, but 3 years later it was in the "dementia" range. The rapidity of the decreases in his mental status and in his ability to perform activities of daily living that occurred when he was admitted to the hospital indicate that, if circumstances permit, elderly men may benefit from living at home with their families. Close attention to diet and exercise from youth through senescence may also contribute to health and longevity. PMID- 9212688 TI - [Tokyo Centenarian Study. 5. Nutritional status of Japanese centenarians]. AB - To establish the guideline for nutrition in the very old, we analyzed biochemical and hematological data from 45 Japanese centenarians living in Tokyo metropolitan area during 1994 and 1995. Levels of cholesterol, HDL-cholesterol, apolipoprotein A1 and B, albumin, prealbumin in serum were lower than in a control group or lower than the reference range, which indicates that these centenarians were undernourished. RBC counts, hemoglobin, and hematocrit were also low in the centenarians. The concentration of albumin correlated positively with those of HDL-cholesterol and apolipoprotein A1, and negatively with concentration of Lp(a). The value for transferrin correlated positively with energy intake and with the concentration of apolipoprotein A1. The concentration of prealbumin correlated positively with the concentration of hemoglobin, hematocrit and the total cholesterol concentration. According to their nutritional status, the centenarians were divided into 2 groups: well-nourished and undernourished. Those who were well-nourished had higher levels of cholesterol, HDL, Hb, and apolipoprotein A1, their levels of ADL and cognitive function were also high. The findings of blood-chemical data in the centenarians may be partly due to undernutrition. Centenarians who were well-nourished were considered to be aging successfully. These results and others previously published indicate that the concentrations of albumin, prealbumin, transferrin and either CRI or IL-6 are useful for nutritional assessment in the very old. PMID- 9212689 TI - [Swallowing rehabilitation in an elderly patient with Wallenberg's syndrome--role of videofluorography and the swallowing provocation test]. AB - A 65-year-old man was admitted to our department due to severe dysphagia, dysarthria, and aspiration pneumonia. Dysphagia and dysarthria were caused by lateral medullary infarction (Wallenberg' s syndrome). After the patient recovered from pneumonia, the abnormality of swallowing was assessed by a swallowing provocation test and videofluorography. Two months after the start of swallowing training, a swallowing provocation test showed that the swallowing reflex had improved and videofluorography showed that the magnitude of aspiration to the trachea had decreased. The patient began taking food by mouth. These tests are useful for quantitative assessment of dysphagia and for deciding when to start oral intake in elderly patients. PMID- 9212690 TI - Nutrition and depression: the role of folate. AB - A relationship between folate and neuropsychiatric disorders has been inferred from clinical observation and from the enhanced understanding of the role of folate in critical brain metabolic pathways. Depressive symptoms are the most common neuropsychiatric manifestation of folate deficiency. Conversely, borderline low or deficient serum or red blood cell folate levels have been detected in 15-38% of adults diagnosed with depressive disorders. Recently, low folate levels have been linked to poorer antidepressant response to selective serotonin reuptake inhibitors. Factors contributing to low serum folate levels among depressed patients as well as the circumstances under which folate and its derivatives may have a role in antidepressant pharmacotherapy must be further clarified. PMID- 9212691 TI - Obesity solutions: report of a meeting. AB - A workshop entitled "Obesity Solutions" was held on January 11, 1996, at St. Luke's-Roosevelt Hospital in New York City and was jointly sponsored by the St. Luke's-Roosevelt Obesity Research Center and the Nestle R&D Center, Inc., of New Milford, Connecticut. The purpose of the workshop was to bring together experts from the research community and the pharmaceutical and food industries to address the epidemic of obesity in the United States and offer potential solutions. The following is a report of that meeting. PMID- 9212692 TI - Effects of alcohol on energy metabolism and body weight regulation: is alcohol a risk factor for obesity? AB - Some studies have suggested that drinking in moderation may be beneficial for health, but many of these studies do not address body weight. Evidence suggests that consuming moderate amounts of alcohol is a risk factor for obesity, which is a risk factor for several adverse health outcomes. Recommendations regarding alcohol intake thus should take into account a variety of factors, including baseline body weight, location of body fat, and overall diet. PMID- 9212693 TI - Scientists' statement regarding data on the sodium-hypertension relationship and sodium health claims on food labeling. AB - Nutrition Reviews has had a continuing interest in the scientific basis of arguments for and against health claims on foods. Such claims were first authorized by the Food and Drug Administration (FDA) in 1993 under the terms of the Nutrition Labelling and Education Act of the U.S. Congress, passed in 1990. Since then there have been petitions that have resulted in a newly authorized claim for oats and other petitions directed toward modification or deletion of the originally approved claims. The Salt Institute, an industry-supported organization, has actively participated in the discussions regarding the sodium and hypertension health claim. The scientists signing this statement are familiar with the evolving understanding of the relationship between sodium and hypertension. The statement reviews more recent studies and critiques the arguments for a sodium health claim. We invite our readers to respond to the discussion in this controversial area of nutrition science and policy. PMID- 9212694 TI - Inositol prevents expression of a genetic model of neural tube defects in mice. AB - Although up to 70% of neural tube defects (NTDs) can be prevented by adequate folate consumption, some NTDs are unrelated to folate, myo-inositol was found to reduce the incidence of spinal NTDs that are unrelated to folate in curly tail mice. PMID- 9212695 TI - A new uncoupling protein: a potential component of the human body weight regulation system. AB - A mitochondrial protein that uncouples the respiratory chain from oxidative phosphorylation and generates heat is found in brown adipose tissue (BAT) of rodents. Although humans have little BAT, a second uncoupling protein has been discovered that is widely distributed in human tissues. It is induced in mice by a high-fat diet and in mice with obese mutations, and may play a role in human body weight regulation. PMID- 9212696 TI - Cloning and the Odyssey. PMID- 9212697 TI - The child with attention deficit hyperactivity disorder and learning disability. AB - The following case is presented to demonstrate the difficulties associated with treatment of Attention Deficit Hyperactivity Disorder (ADHD) when it is comorbid with a learning disability (LD). These two disorders exist simultaneously in about 30% of those diagnosed with ADHD. It is often difficult to separate the effects of one condition from those of the other. Researchers and clinicians often struggle with questions concerning the origin of inattentive symptoms. A child with LD may have trouble attending if placed in a classroom setting not designed to deal with issues related to LD, while a child with attention deficit may not be progressing as well as expected because of problems maintaining attention. This case highlights the need for a multi-faceted approach to the treatment of ADHD, as well as, the need for continued evaluation of the effectiveness of the treatment plan instituted. PMID- 9212698 TI - Herbal remedies. PMID- 9212699 TI - Expanded programme on immunization (EPI) measles and the interruption of indigenous transmission, 1996. PMID- 9212700 TI - Food safety. Enterohaemorrhagic Escherichia coli (EHEC) infection. PMID- 9212701 TI - Atomic and molecular imaging at the single-cell level with TOF-SIMS. AB - A complete cold chain freeze-fracture methodology has been developed to test the feasibility of using time-of-flight secondary ion mass spectrometry (TOF-SIMS) imaging for the molecular analysis of frozen hydrated biological samples. Because the technique only samples the first few monolayers of a sample, water on the surface of a sample can be a major source of interference. This problem can be minimized by placing a cold trap (fracture knife and housing at -196 degrees C) near the fractured sample that is held at a warmer temperature (-97 to -113 degrees C). This results in removal of surface water and prevents condensation on the surface. Although this approach is effective, it has been found that sample warming needs to be carefully controlled due to the volatility of other matrix molecules and the morphological effects imparted onto the cell surface during drying. By utilizing the above handling technique, it has been possible to demonstrate for the first time that TOF-SIMS imaging technology can be used to obtain images of molecular species across a cell surface with a submicrometer ion probe beam. Images of small hydrocarbons and the deliberately added dopants DMSO and cocaine have been obtained with TOF-SIMS of the single-cell organism Paramecium. PMID- 9212702 TI - Two-dimensional SEC/RPLC coupled to mass spectrometry for the analysis of peptides. AB - A two-dimensional liquid chromatography system is described here which uses size exclusion liquid chromatography (SEC) followed by reversed phase liquid chromatography (RPLC) to separate the mixture of peptides resulting from the enzymatic digestion of a protein. A novel LC/LC interface, using two RPLC columns in parallel rather than storage loops, joins the two chromatographic dimensions. This new interface design permits the use of conventional analytical diameter HPLC columns, 7.8 mm for SEC and 4.6 mm for RPLC, making construction and maintenance of this system very easy. The reversed phase chromatography utilizes 1.5 microns diameter, nonporous C-18 modified silica particles, which produce fast and efficient analyses. Following the high-resolution two-dimensional chromatographic separation, an electrospray mass spectrometer detects the peptide fragments. The mass spectrometer scans a 2000 m/z range to identify the analytes from their molecular weights. The analyses of tryptic digests of ovalbumin and serum albumin are each described. PMID- 9212703 TI - Ultrathin slab gel separations of DNA using a single capillary sample introduction system. AB - We demonstrate here a novel method for DNA separations which combines the parallel processing capabilities of slab gels with the advantages of sample introduction obtained with a single capillary. This sample introduction format allows rapid sequential separations or continuous analysis to be carried out on ultrathin slab gels with efficient heat dissipation. Ultrathin slab gels have been fabricated by using 57-micron spacers between quartz plates, and a single capillary has been used to introduce plugs of dsDNA fragments into the ultrathin gel. These fragment plugs were deposited along the entrance to the ultrathin gel at spatially discrete locations by micromanipulation of the capillary. Spatially resolved detection has been accomplished with an argon ion laser focused to a line for excitation and a CCD for collection of fluorescence. Double-stranded DNA separations are demonstrated in a plug injection format. This approach allows multiple unique samples to be rapidly deposited on the ultrathin slab gels for separation. PMID- 9212704 TI - Confirmation of the structure of lipid A derived from the lipopolysaccharide of Rhodobacter sphaeroides by a combination of MALDI, LSIMS, and tandem mass spectrometry. AB - The chemical structure of nontoxic diphosphoryl lipid A from Rhodobacter sphaeroides was confirmed using a combination of LSIMS (on a two-sector mass spectrometer) and MALDI (on time-of-flight and ion trap mass spectrometers) in conjunction with tandem mass spectrometry in both positive and negative ion modes. Accurate molecular weight measurement accompanied by the analysis of fragment ion masses yielded the composition of fatty acyl groups. Tandem experiments (collisionally induced dissociation of both quasimolecular and oxonium ions) were also performed, revealing the precise location and nature of the fatty acyl groups on the disaccharide backbone. PMID- 9212705 TI - Characterization of immobilization of an enzyme in a modified Y zeolite matrix and its application to an amperometric glucose biosensor. AB - A new approach to construct an amperometric biosensor is described. Without using bovine serum albumin-glutaraldehyde, glucose oxidase (GOx) was immobilized on a dealuminized Y zeolite (DAY)-modified platinum electrode to construct a glucose sensor. The large specific surface area of the zeolite substrate resulted in high enzyme loading. The immobilized GOx in this manner was stable and could maintain its high activity for at least 3 months. The interactions between the zeolite and the enzyme were investigated by means of Fourier transform infrared spectra, and the pore distribution and the surface acid property of DAY were preliminarily studied. The results showed that the hydrophilic property and the existing mesopores of DAY played important roles in the enzyme immobilization. This resulting biosensor exhibited good reproducibility and selectivity, owing to the uniform pore structure and unique ion-exchange property of the zeolite. The biosensor responded rapidly to glucose in the linear range from 2.0 x 10(-6) to 3.0 x 10(-3) M, with a detection limit of 0.5 microM. PMID- 9212706 TI - Characterization of visible dyes for four-decay fluorescence detection in DNA sequencing. AB - Dyes of several classes were investigated as candidates for use in a multiplex, four-decay fluorescence detection scheme for DNA sequencing. The dyes include nitrobenzofuran dyes, rhodamine dyes, fluorescein dyes, cyanine dyes, Nile Red, and BODIPY dyes. Based on the results of fluorescence spectral and lifetime studies, an initial set of four dyes was selected for further study: NBD aminohexanoic acid (NBD-HA, r = 1.1 ns), tetramethyl-rhodamine, methyl ester (r = 2.2 ns), rhodamine green (r = 4.3 ns), and BODIPY 505/515 (r = 5.9 ns). Limits of lifetime detection of the four dyes were investigated, and lifetime resolution was demonstrated for mixtures of the free dyes in batch solution. Lifetime of dye labeled DNA primers also were determined in batch solution and detected on-the fly in capillary electrophoresis (CE). Conjugation of the dyes to DNA improved the resolution of their individual lifetimes in mixtures in batch measurements. When attached to the primer, tetramethyl-rhodamine exhibited biexponential decay with a dominant lifetime of 3.8 ns, making it unsuitable for four-decay sequencing. Contact with the CE gel lengthened the lifetime of NBD-HA-labeled primer from 1.3 to 2.1 ns but did not affect the lifetimes of the other dyes. Lifetime detectability of labeled primers at individual points along an electrophoretic peak in the attomole range. PMID- 9212707 TI - Use of a polybrene capillary coating in capillary electrophoresis for rapid analysis of hemoglobin variants with on-line detection via an ion trap storage/reflectron time-of-flight mass spectrometer. AB - A polybrene capillary coating in capillary electrophoresis (CE) has been used for rapid analysis of hemoglobin variant digests. The use of the polybrene capillary coating has allowed sufficient separation to resolve the large number of digest products formed upon tryptic digestion of the whole protein, so that prior separation of the hemoglobin alpha and beta chains is not required. The resolution of the digest peaks obtained by CE is sufficient so that even single amino acid substitutions can easily be detected using UV absorption detection. The digest is further analyzed by capillary electrophoresis separation with on line detection using electrospray ionization interfaced to the ion trap storage/reflectron time of flight device (CE/ESI-IT/reTOF), where a comparison of the total ion electropherograms and mass spectra of the mutant and normal hemoglobins can detect the presence of a mutation site. The CE separation and mass analysis can be accomplished in typically 10-15 min. The unique capability of the CE/ESI-IT/reTOF system for detection of fast separations with narrow peaks that may be under 1 s fwhm is demonstrated. The speed of this system is essential for resolution of the large number of peaks that are separated in a short time duration using CE separations. PMID- 9212708 TI - Characterization of the glycation of albumin in freeze-dried and frozen human serum. AB - Human serum albumin (HSA) in fresh frozen and freeze-dried serum reference materials was examined by mass spectrometry and a variety of affinity chromatography techniques. The relative molecular mass distribution of HSA in fresh frozen serum was found to be identical to that of an HSA standard. However, the HSA in the freeze-dried reference serum exhibited a relative molecular mass distribution that was shifted to higher mass, broader, and substantially more heterogeneous than that of HSA in fresh frozen serum. A proteolytic cyanogen bromide digestion of the HSA from freeze-dried serum contained adducts approximately 162 u higher in mass than digest fragments 124-298 and 447-548, suggesting glycation. The presence of glycation on fragments 124-298 and 447-548 correlates with the known sites of HSA glycation. Glycation was further confirmed by the mass spectral analysis of the retained and unretained fractions from glycoaffinity chromatography of HSA from freeze-dried serum. The relative molecular weight of the HSA in the retained fraction indicated the presence of a doubly glycated species. The chemical heterogeneity of Cys-34, the site of the only free thiol in HSA, was examined and found not to be a substantial source of molecular mass heterogeneity for HSA from either fresh frozen of freeze-dried serum. PMID- 9212709 TI - Direct analysis of sugar alcohol borate complexes in plant extracts by matrix assisted laser desorption/ionization fourier transform mass spectrometry. AB - Matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI/FTMS), operating in the negative ion mode, is used to directly observe sugar alcohol borate complexes in a number of plant fractions. The method involves virtually no sample workup and, in the case of celery phloem sap, requires only 40 nL of sap to observe the borate complex. The isolation and characterization of such soluble borate complexes is important in understanding the distribution of boron in plants. The results show that the complexes are composed of two mannitol or sorbitol ligands (L) complexed to a single borate center (B). In some cases, the boron is complexed to non-alditol monosaccharides. Sustained off-resonance collision irradiation dissociation of the BL2- complex, where L is a mannitol, gives fragments that confirm the proposed structure. Complexes of larger oligosaccharides have also been successfully observed using MALDI/FTMS. Semiempirical molecular orbital calculations (AM1) of the mannitol BL2- complex show that the most favorable configuration is with carbons 3 and 4 of both mannitol residues complexed to the borate. This allows maximum interaction of the remaining hydroxyls with the borate center. PMID- 9212710 TI - Quantitative comparison of global carbohydrate structures of glycoproteins using LC-MS and in-source fragmentation. AB - A comparative method for the quantitative analysis of the ratio of oxonium fragment (reporter) ions derived from sialic acid and N-acetylhexosamine residues on a large intact glycoprotein, the B domain of recombinant human factor VIII (rhFVIII), was developed. The method utilized liquid chromatography-electrospray ionization mass spectrometry (LC-ESI MS) on a single-quadrupole instrument. During development, systematic approaches such as full-matrix and simplex strategies were used for the optimization of the signal-to-noise ratio by controlling source temperature and cone voltage. The method was found to be precise (RSD = 0.84%), sensitive (capable of differentiating 1 sialic acid residue change among at least 29 sialic acids on a 103-kDa glycoprotein that is 38% carbohydrate), applicable to a wide range of loading (11.6-372 micrograms of FVIII), and accurate according to a comparison to matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. Combining the method with enzymatic removal of N-glycans, selective O-glycan analysis was also performed leading to differential fragment ion analysis ascribed to N- and O-glycans. Quantitative ESI in-source dissociation MS combined with LC can generally be used for glycoproteins, as one of the indicators, to compare the distribution of carbohydrate residues over N- and O-glycans, to investigate their isoforms, and compare batch-to-batch characteristics of biopharmaceuticals. PMID- 9212711 TI - A MALDI probe for mass spectrometers. AB - A new MALDI probe has been designed that uses transmission geometry. This geometry allows the probe to be fashioned after typical EI/CI solid probes which enables it to be introduced into spatially constrained ion source regions such as encountered in quadrupole ion trap mass spectrometers. In the probe design demonstrated here, light from a fiber optic irradiates the backside of a sample through a small piece of quartz on which the sample has been directly deposited. The performance characteristics exhibited by utilizing this probe for MALDI on a quadrupole ion trap mass spectrometer are similar to those which can be obtained through the traditional methods of implementing MALDI. Spectra have been obtained from 50 fmol of total loading of bombesin, MS/MS has been performed on 5 pmol of des-Arg9-bradykinin, and the analyte ion signal is shown to last for over 2500 laser shots for 2 pmol of bombesin. Optical micrographs showing the crystal distribution of a sample containing 2 pmol of bombesin have been obtained as a function of the number of laser shots for a single sample loading. Although this probe was designed for use with the quadrupole ion trap, it can be adapted for use with all types of mass spectrometers. Thus, with only one laser, one fiber optic, and this probe, MALDI can be performed on multiple instruments in a lab. PMID- 9212712 TI - Accuracy of empirical correlations for estimating diffusion coefficients in aqueous organic mixtures. AB - Diffusion coefficients for a homologous series of alkylbenzenes and alkylphenones have been measured by the Aris-Taylor open tube method from 30 to 60 degrees C over a wide range in methanol/water and acetonitrile/water compositions (10-100% by volume of organic). The measurements were compared to estimates derived from five of the most common empirical correlations. The errors for methanolic mixtures by the Wilke-Chang, Scheibel, and Lusis-Ratcliff correlations are usually less than 20%. The Scheibel, Wilke-Chang, and Hayduk-Laudie correlations work better than others for acetonitrile/water mixtures. Overall, the Scheibel correlation shows the smallest errors, and we recommend its use to that of the more widely used Wilke-Chang method for the systems studied here. We have also developed fitting equations for estimating viscosity so that the diffusion coefficients can be easily estimated. PMID- 9212713 TI - Estimating diffusion coefficients for alkylbenzenes and alkylphenones in aqueous mixtures with acetonitrile and methanol. AB - The accuracy of various empirical approximation methods of estimating the diffusion coefficients of alkylbenzenes and alkylphenones in acetonitrile (ACN)/water and methanol (MeOH)/water solvent systems is reported. Diffusion coefficients for these solutes have been measured over a wide range of solvent compositions and temperatures. A novel empirical modification of the Wilke-Chang method has been developed by correlating measured values with solute and solvent parameters. The correlation, along with the Wilke-Chang and Scheibel correlations, was examined by comparing the computed diffusion coefficients with the measured values. We find that the percent errors of the proposed correlations are no greater than 10% for both ACN/water and MeOH/water systems and that the accuracy of the correlation is 2-3-fold better than those of the other two correlations. We recommend the use of this correlation with the above homologous series of solutes for the evaluation of column performance in reversed-phase liquid chromatography. PMID- 9212715 TI - Effect of tissue and plasma factors on kidney proliferation. AB - Liver extract, plasma from intact mice, ES2 tumour extract and plasma from tumour bearing mice has an inhibiting effect on the mitotic activity of hepatocytes and duodenal enterocytes. In the present experiments, the effect of these treatments on the mitotic activity of renal tubular cells was studied. C3HS 28 day-old male mice, standardized for periodicity analysis were used. The determination of normal mitotic circadian curve of the renocytes was done. A second batch of mice were injected with 0.01 ml/gr of either liver extract, plasma from intact mice, ES2 tumour extract or plasma from tumour bearing mice, at 16:00 hours and controlled at 08:00, 12:00 and 16:00 hs during 2 consecutive days post treatment. Colchicine (2 micrograms/gr) was injected 4 hours before killing. Kidneys were processed for histology and mitotic index determinations. Results were expressed as colchicine metaphases per 1000 nuclei, and showed that mitotic activity values of treated animals were significantly lower than those of controls. In conclusion, mitotic activity inhibition of renocytes may be due to some non specific plasmatic and/or tissue factors. PMID- 9212714 TI - Fiber-optic-based biomonitoring of benzene derivatives by recombinant E. coli bearing luciferase gene-fused TOL-plasmid immobilized on the fiber-optic end. AB - TOL plasmid in Pseudomonas putida mt-2 has a series of genes for the degradation of xylene, toluene, and their derivatives to pyruvate and acetaldehyde (or propionaldehyde). Two operons, i.e., upper operon and meta operon, play indispensable roles for the digestion of xylene derivatives: When XyIR protein recognizes xylene derivatives, another controlling gene, xyIS, is activated, which results in the activation of meta operon. Therefore, we have constructed a fusion gene between TOL plasmid and the firefly luciferase gene under the control of XyIR and the promoter of xyIS gene; i.e., by using fusions of the meta operon with promotorless luciferase expression vector from firefly, we have constructed and tested biomonitors for benzene derivatives. Bioluminescence specified by Escherichia coli (pTSN316), carrying xyIR and xyIS promoters, Ampr and luc, was measured in either a benzene derivative-saturated or o-methylbenzyl alcohol dissolved medium both in the case of cell suspension and in the case of immobilized cell form. The utility of the biosensing system for monitoring in chemical plant drainage was demonstrated with samples supplemented with benzene derivatives. The xyIR-xyIS promoter-lux fusion carried by pTSN316 responded to a benzene-related chemical in sample solutions. Immobilization of the transformed E. coli, at one end of fiber optic, bearing firefly luciferase gene fused to TOL plasmid, has been demonstrated to fabricate a luminescent remote biomonitoring device for the protection of environmental deterioration. Due to the luminescent detection, the detection limits for benzene-related aromatics that are recognized by a binding protein (XyIR) were parts-per-million. We had already submitted a preliminary report concerning the possibility of environmental monitoring based on the above idea by using the transformed E. coli in a cell-suspended solution. This paper describes mainly a fiber-optic-based biomonitoring device for the protection of environmental deterioration. PMID- 9212716 TI - Seasonal and experimental reactivation of Leydig cells of the bat Tadarida brasiliensis. AB - The Leydig cells of the bat Tadarida brasiliensis, exhibit two well-defined periods of secretory activity that are intimately associated to the bat reproductive cycle. During the breeding season (August-September, late Winter and early Spring in the southern hemisphere), the interstitial tissue contains hypertrophic Leydig cells characterized ultrastructurally by the presence of pleomorphic mitochondria, depletion of lipid droplets, proliferation of membranes of agranular endoplasmic reticulum (AER) and enlargement of the Golgi complexes. By contrast, from Spring to Fall concurrent with regression of seminiferous tubules, the Leydig cells acquire a quiescent appearance with reduction in size and volume of AER membranes, atrophy of the Golgi complex and a massive storage of lipid droplets. The changes occurring in Leydig cells during the breeding season can be duplicated experimentally in non-breeding bats with exogenous stimulation with hCG. The gonadotropic treatment induces rapid changes in both interstitial cells and seminiferous tubules. The latter present evident signs of reactivation including proliferation of the spermatogenic cell line, permeation of the tubular lumen and depletion of lipid droplets. The Leydig cells display similar features to those found in the bat during the mating season at the peak of secretory activity. The bat T. brasiliensis is an excellent model to correlate the morphological organization of the Leydig cells with either seasonal fluctuations of its secretory activity or after experimental stimulation with gonadotropins. PMID- 9212717 TI - Detection of endothelial cells by MEC 13.3 monoclonal antibody in mice mammary tumors. AB - In order to clearly visualize blood vessels, the monoclonal antibody (mAb) MEC 13.3 was used for an immunohistochemical staining on frozen sections of different mice mammary tumors. MEC 13.3 mAb is specific for endothelial cells (ECs) of mouse blood vessels and recognizes a molecule related to the murine form of CD31/PECAM. This mAb with immunoenzymatic technique or immunofluorescent labelling, was found to be a useful tool to quantify tumor neovascularization. Specifically, membrane reinforcement could be observed in vessel ECs, indicating the expression of CD31/ PECAM in their surface. The staining of ECs from tumors and from normal tissues was also compared. In this work, the use of MEC13.3 mAb is reported to recognize mice mammary tumor ECs as a useful tool to identify neovascularization. It would also be helpful for research on the origin and function of vascular endothelium in murine tumor experimental models. PMID- 9212718 TI - Glucocorticoid regulation of in vitro astrocyte phagocytosis. AB - Astrocytes participate in central nervous system injury, degenerative diseases and also perform macrophagic functions. The present work investigates: 1) the effect of the physiological glucocorticoid corticosterone (CORT) and the synthetic agonist dexamethasone (DEX) on latex beads phagocytosis by neonatal rat cortical astrocytes in culture, and 2) the expression of immunoreactive glucocorticoid receptors (GR) in astrocytes cultured in different media with or without a pulse application of CORT. The results indicated that glucocorticoids reduced astrocyte phagocytic activity, as occurred with macrophages, independently of the culturing conditions employed. The extent of phagocytosis was inversely related to nuclear immunostaining for GR in cultures in fetal calf serum, which contained endogenous glucocorticoid. However, no correlation was found between nuclear GR and phagocytosis for cultures in glucocorticoid-free medium or in medium containing CORT. It is suggested that additional factors, besides the GR, may be involved in glucocorticoid modulation of astrocyte phagocytosis. PMID- 9212719 TI - A technique for chronically recording the EKG of the lymph hearts in the toad Bufo arenarum (Hensel). AB - A simple technique for recording the EKG of the posterior lymphatic hearts of the toad Bufo arenarum (Hensel) in free moving unanesthetized specimens is described. This technique permits long term chronic recordings in varied physiological and behavioral conditions whereas it overcomes some of the technical difficulties encountered in obtaining reliable recordings. PMID- 9212720 TI - Working memory: a view from neuroimaging. AB - We have used neuroimaging techniques, mainly positron emission tomography (PET), to study cognitively driven issues about working memory. Two kinds of experiments are described. In the first kind, we employ standard subtraction logic to uncover the basic components of working memory. These studies indicate that: (a) there are different working-memory systems for spatial, object, and verbal information (with the spatial system localized more in the right hemisphere, and the verbal system more in the left hemisphere); (b) within at least the spatial and verbal systems, separable components seem to be responsible for the passive storage of information and the active maintenance of information (with the storage component being localized more in the back of the brain, and the maintenance component in the front); and (c) there may be separate components responsible for processing the contents of working memory (localized in prefrontal cortex). In our second kind of experiment we have focused on verbal working memory and incrementally varied one task parameter-memory load-in an effort to obtain a more fine-grained analysis of the system's operations. The results indicate that all relevant components of the system show some increase in activity with increasing memory load (e.g., the frontal regions responsible for verbal rehearsal show incremental increases in activation with increasing memory load). In contrast, brain regions that are not part of the working-memory system show no effect of memory load. Furthermore, the time courses of activation may differ for regions that are sensitive to load versus those that are not. Taken together, our results provide support for certain cognitive models of working memory (e.g., Baddeley, 1992) and also suggest some distinctions that these models have not emphasized. And more fundamentally, the results provide a neural base for cognitive models of working memory. PMID- 9212722 TI - Bridging the gap between monkey neurophysiology and human perception: an ambiguity resolution theory of visual selective attention. AB - When the visual system must process multiple objects simultaneously, as in the visual search paradigm, the neural coding of individual objects can become ambiguous due to the visual system's extensive use of coarse coding and distributed representations. Here we propose that the primary role of visual selective attention within the ventral object recognition pathway is to resolve these ambiguities. We begin by reviewing previous studies of the effects of attention on neural responses in monkeys, which provide the basis for this hypothesis, and then describe a new set of experiments showing that similar attentional mechanisms operate in the human brain. In these new experiments, event-related potentials (ERPs) were recorded from normal human observers while they performed tasks analogous to those used previously in monkeys. The central finding was that an attention-related ERP wave called the "N2pc component" was present under the same conditions that led to attentional modulations of neural responses in monkey visual cortex. These human electrophysiological results provide a bridge between cognitive-level theories of visual attention and the behavior of individual neurons in visual cortex. PMID- 9212721 TI - Neural substrates of fluid reasoning: an fMRI study of neocortical activation during performance of the Raven's Progressive Matrices Test. AB - We examined brain activation, as measured by functional magnetic resonance imaging, during problem solving in seven young, healthy participants. Participants solved problems selected from the Raven's Progressive Matrices Test, a test known to predict performance on a wide range of reasoning tasks. In three conditions, participants solved problems requiring (1) analytic reasoning; (2) figural or visuospatial reasoning; or (3) simple pattern matching that served as a perceptual-motor control. Right frontal and bilateral parietal regions were activated more by figural than control problems. Bilateral frontal and left parietal, occipital, and temporal regions were activated more by analytic than figural problems. All of these regions were activated more by analytic than match problems. Many of these activations occurred in regions associated with working memory. Figural reasoning activated areas involved in spatial and object working memory. Analytic reasoning activated additional areas involved in verbal working memory and domain-independent associative and executive processes. These results suggest that fluid reasoning is mediated by a composite of working memory systems. PMID- 9212723 TI - Cellular and molecular mechanisms of disease: a 5-year cycle in the long life of The American Journal of Pathology. PMID- 9212724 TI - Pathology: a historical opportunity. PMID- 9212725 TI - Discovery of new lesions in neurodegenerative diseases with monoclonal antibody techniques: is there a non-amyloid precursor to senile plaques? AB - The manuscript by Schmidt et al reports that antibodies generated to paired helical filaments (AMY antibodies) unexpectedly labeled novel non-amyloid, plaque like structures (AMY plaques) in aged and Alzheimer's disease brains. The full disclosure of the nature of these lesions awaits additional structural and biochemical studies, but at first glance there are interesting parallels between AMY plaques and recently described lesions composed of glia and glia-associated proteoglycans in brains of aged mice. The increasing recognition of the role of proteoglycans in paired helical filaments formation makes proteoglycans or their associated molecules attractive candidates for AMY-immunoreactive proteins. The relationship of AMY plaques to age-related glial changes that some have speculated may be precursors to senile plaques remains to be determined, as is the relationship of AMY plaques to more widely recognized amyloid-containing plaques. Future studies will determine whether AMY plaques are non-amyloid precursors to senile plaques or if they represent an independent type of structural lesion in the Alzheimer's disease brain. Ultimately, the clinical significance of AMY plaques will depend upon their characterization in brains of prospectively studied subjects. PMID- 9212726 TI - Vascular endothelial growth factor and ocular neovascularization. AB - Okamoto et al have developed an extremely useful and interesting model of retinal and subretinal neovascularization. Using molecular techniques, they have developed a transgenic model driven by overexpression of VEGF, a growth factor demonstrated to play an important role in neovascularization in many ocular diseases. They have been able to demonstrate that VEGF overexpression is sufficient to cause intraretinal and subretinal neovascularization. The mouse model is relatively cheap and reliable, does not require any exogenous agent, and has many characteristics of clinical intraocular neovascularization. The new vessels develop in the outer retina and subretinal space and have a characteristic histological appearance. They leak fluorescein on angiography, demonstrating their similarity to human disease and allowing identification and grading of neovascularization in vivo. The model can be used to investigate molecular mechanisms of VEGF-dependent neovascularization, with applications beyond ocular eye disease. The model can also be used to study anti-angiogenic agents that have the potential to treat common blinding diseases such as age related macular degeneration. Okamoto et al have made a substantial contribution to the angiogenesis field with this work, and one looks forward to future investigations. PMID- 9212728 TI - Transforming growth factor-beta receptor type 2 mutations and microsatellite instability in sporadic colorectal adenomas and carcinomas. AB - Frame-shift mutations in a run of 10 adenines (A10) of the transforming growth factor-beta receptor type 2 gene (TGF-beta RII) are commonly seen in inherited and sporadic colonic cancers that exhibit microsatellite instability. A10 mutations and instability also are commonly seen in hereditary nonpolyposis colon cancer-associated adenomas. However, instability is quite rare in sporadic adenomas, and the timing of acquisition of A10 mutations with respect to the sporadic adenoma-carcinoma sequence has not been reported. We evaluated 100 sporadic colorectal cancers and 164 sporadic adenomas for microsatellite instability with a set of 10 tetranucleotide polymerase chain reaction primer sets and for A10 frame-shift mutations. A10 mutations were significantly associated with microsatellite instability in colorectal cancers, being seen in 9 of 11 cancers with 50% or greater instability and in 0 of 89 tumors with less than 50% instability (P < 0.0001). A10 mutations were not detected in any adenomas, only three of which (1.8%) exhibited significant (30% or greater) instability. We conclude that both TGF-beta RII frame-shift mutations and microsatellite instability occur at a relatively late stage of sporadic colorectal tumorigenesis. A10 frame-shift mutations appear to be restricted to sporadic colorectal cancers with extensive microsatellite instability. PMID- 9212727 TI - Telomerase activation in cervical cancer. AB - It has been hypothesized that infection with high-risk human papillomaviruses (HPVs), in conjunction with other cellular events, plays a critical role in the development of cervical cancer. Activation of telomerase, a ribonucleoprotein enzyme complex that synthesizes telomere repeats, has been associated with acquisition of the immortal phenotype in vitro and is commonly observed in human cancers. In this study, we have examined 10 high-grade cervical cancers for telomerase activity and for the presence of HPV. Telomerase activity was detected in all of the cancers but in none of the paired histopathologically normal uterine tissues or in normal cervical epithelium. Analysis of these same tissues for HPV nucleic acids by polymerase chain reaction (PCR) using primers from the HPV L1 and E6 open reading frames demonstrated that 7 of 10 cancers were positive for HPV, 3 for HPV type 16 (HPV-16), and 4 for HPV-18. In one case, HPV-16 was detected in histopathologically normal uterine tissue, the same type as that detected in the cancer from the same patient. HPV DNA was not detected in 3 of 10 cancers. These results indicate that telomerase activation is common in high grade cervical cancers and suggests that telomerase activity may be a useful diagnostic marker for the disease. PMID- 9212729 TI - Expression of HMGI-C and HMGI(Y) in ordinary lipoma and atypical lipomatous tumors: immunohistochemical reactivity correlates with karyotypic alterations. AB - The high mobility group proteins (HMGs) are a class of low molecular weight, nonhistone, nuclear proteins that bind DNA and function as transcription cofactors. This class includes the HMGI family members HMGI-C and HMGI(Y). Both are not significantly expressed in differentiated adult tissues, including fat, but their expression is induced in proliferating and transformed cells. Their involvement in the development of lipomatous tumors has been recently demonstrated for HMGI-C, which is encoded by a gene located at 12q15, the chromosomal segment often rearranged in ordinary lipomas. The same chromosomal segment is consistently amplified in the ring and giant marker chromosomes of atypical lipomatous tumors (ALTs), a term used to designate tumors previously labeled as well differentiated liposarcomas or atypical lipomas. The involvement of HMGI(Y) is strongly suspected as the gene coding for HMGI(Y) is located at 6p21, a chromosomal segment rearranged in a subset of ordinary lipomas. HMGI-C or HMGI(Y) protein expression was analyzed immunohistochemically in a group of 39 well differentiated adipose neoplasms (19 lipomas and 20 ALTs) of known karyotype using polyclonal antibodies raised against a recombinant protein (HMGI-C) and against a synthetic peptide (HMGI(Y)). The results of this study demonstrate that HMGI proteins are commonly expressed in well differentiated adipose neoplasms. Seventeen of twenty ALTS (85.0%), all of which had ring or giant marker chromosomes with amplification of 12q13-15, strongly expressed HMGI-C. HMGI-C expression was detected in 7 of 11 ordinary lipomas (63.6%) with alterations at 12q14-15 and in one case with an abnormal karyotype that included double minute chromosomes. HMGI-C immunoreactivity correlates with 12q13-15 chromosomal alterations (P = 0.001). HMGI(Y) reactivity was demonstrated in only two ordinary lipomas: one with 6p21 rearrangement and one with normal karyotype. No significant HMGI(Y) expression was found in the ALT group. The finding of aberrant expression of HMGI proteins in well differentiated adipose neoplasms in association with 12q13-15 and 6p21 chromosomal changes supports the proposed pathogenetic role of this group of proteins in the development of adipose tissue tumors. PMID- 9212730 TI - Expression of occludin, tight-junction-associated protein, in human digestive tract. AB - The tight junction seals cells together at a subapical location and functionally separates the plasma membrane into an apical and a basolateral domain. This junction is one of the most characteristic structural markers of the polarized epithelial cell. Recently, occludin has been identified as an integral transmembrane protein localizing at the tight junction and directly associated with ZO-1, an undercoat-constitutive cytoplasmic protein. We have investigated occludin expression in conjunction with ZO-1 in normal epithelia and cancers of human digestive tract by immunostaining with a new antibody raised against human occludin. In the normal simple columnar epithelium, occludin was expressed together with ZO-1 as a single line at the apical cell border. However, in the esophagus, which has a stratified squamous epithelium, no occludin expression could be detected, but ZO-1 was expressed in the spinous layer. As for cancers, both occludin and ZO-1 showed the same expression in differentiated adenocarcinoma cells as in normal epithelium, but in poorly differentiated adenocarcinomas, the expression of these two proteins was reduced. There was significant correlation between tumor differentiation and expression of these proteins. These results suggest that occludin, together with ZO-1, is involved in the formation of gland-like structures. In addition, occludin expression can serve as a histopathological indicator for differentiation in gastrointestinal adenocarcinomas. PMID- 9212732 TI - Microbeam MOMeNT: non-contact laser microdissection of membrane-mounted native tissue. AB - The analysis of tissue-specific genetic alterations depends on the selective procurement of homogeneous cell populations. Microbeam microdissection of membrane-mounted native tissue (MOMeNT) permits the rapid, selective, and low contamination procurement of tumor or other cells from histological sections by non-thermic non-contact laser microdissection. Tissue sections are mounted on a specifically designed ultrathin transparent supporter membrane. Tissue together with the membrane are then dissected with an ultraviolet (337-nm) pulsed laser microbeam coupled into a robot-stage microscope. The ultraviolet laser causes dissection by cold photolysis due to the high photon density of the microbeam rather than by local heating. The track of the laser microbeam can be preselected freely on a video screen, and the size and form of the dissectates can thus be adapted to the histological features of the section with a delineation accuracy in the micron range. Polymerase chain reaction amplification of DNA from the dissectates is not impaired, and tumor-specific loss of heterozygosity of the APC gene as well as homozygous deletion of the MTS1 gene are demonstrated in bladder carcinomas. Taken together, microbeam MOMeNT is a novel technique that utilizes membrane-based microdissection by an ultraviolet laser microbeam, thus providing a flexible, easy-to-use high-performance tool for the molecular pathologist. PMID- 9212731 TI - Immunohistochemical and in situ hybridization detection of growth-hormone producing cells in human thymoma. AB - We have studied 25 thymomas by both immunohistochemistry and in situ hybridization for the presence of growth hormone (GH)-producing cells. Our results indicate that 1) GH-immunoreactive cells were present in 13 of 17 thymomas of cortical and predominantly cortical type but not in medullary (spindled) thymomas (n = 3) or low- to high-grade thymic carcinomas (n = 5), 2) GH-positive cells were mainly located at the periphery of the neoplastic lobules, at the periphery of the perivascular spaces and in the areas of medullary differentiation, 3) cells containing GH mRNA appeared at locations similar to those of GH-immunoreactive cells, and 4) GH-immunoreactive material was present only in the epithelial cell component as revealed by immunoelectron microscopy. In conclusion, this paper demonstrates the occurrence of GH-producing cells in noncarcinoid thymic tumors. The relevance of GH in thymoma cell biology requires additional investigations. PMID- 9212733 TI - Monoclonal antibodies to a 100-kd protein reveal abundant A beta-negative plaques throughout gray matter of Alzheimer's disease brains. AB - Here we describe the initial characterization of a 100-kd protein recognized by four new monoclonal antibodies that reveal abundant and unique plaque-like lesions throughout gray matter of Alzheimer's disease brains. This 100-kd protein and these new plaque-like lesions were identified by four monoclonal antibodies raised to immunogens extracted from Alzheimer's disease neurofibrillary abnormalities. However, these antibodies did not recognize hyperphosphorylated tau in Western blots or neurofibrillary lesions by immunohistochemistry. As all of these antibodies displayed similar properties, one, AMY117, was used to characterize the new plaque-like lesions in detail. These studies demonstrated that AMY117-positive plaques were not visualized by amyloid stains and never co localized with A beta deposits, although AMY117-positive and A beta-positive lesions frequently occurred in the same cortical and subcortical gray matter regions. Abundant AMY117-positive plaques were found in the brains of all 32 sporadic Alzheimer's disease patients and all 6 elderly Down's syndrome subjects. Although AMY117-positive plaques also were seen in the brains of nondemented patients with numerous A beta deposits. AMY117-positive plaques were rare or absent in the brains of other elderly controls and patients with other neurodegenerative or neuropsychiatric disorders. We conclude that the AMY117 positive plaques described here for the first time are major lesions of the Alzheimer's disease brain. Thus, it will be important to elucidate the role played by the 100-kd protein and the AMY117 plaques in the etiology and pathogenesis of Alzheimer's disease. PMID- 9212734 TI - Signal transducer and activator of transcription 1 in human muscle: implications in inflammatory myopathies. AB - Polymyositis (PM) and dermatomyositis (DM) are two major and distinct inflammatory myopathies. Cytokines, implicated in the immune process, have been recognized in the muscle tissue from PM and DM patients, but their functional in situ role has not been identified. We analyzed the expression of the signal transducer and activator of transcription 1 (STAT1), a molecule whose up regulation indicates the interaction of cytokines, or growth factors, with their target receptors in muscle fibers and inflammatory infiltrates in PM and DM. An immunohistochemical analysis was performed using monoclonal antibodies to STAT1 in 57 muscle biopsies from 10 patients with DM, 10 with PM, and 37 controls. The profile of STAT1 up-regulation was also investigated in cultured muscle stimulated by interferon-gamma, epidermal growth factor, platelet-derived growth factor, and interleukin-2, using semiquantitative polymerase chain reaction and Western blot. High STAT1 expression was observed in many perifascicular atrophic muscle fibers from DM patients in 10/10 biopsies. In contrast, only a few muscle fibers undergoing necrosis were STAT1 positive in 2/10 patients with PM and in 2/37 controls. STAT1 reactivity was noted in most cells of the infiltrates in DM, PM, and controls. In vitro, STAT1 was stimulated by interferon-gamma but not by the other molecules studied. These results suggest that in DM, but not in PM, there is distinctive functional local cytokine activity able to increase STAT1 expression in muscle fibers. As interferon-gamma specifically activates STAT1 in vitro, this cytokine in conjunction with ischemia is probably involved in perifascicular muscle fiber pathology in DM. PMID- 9212735 TI - Active replication of HIV-1 at the lymphoepithelial surface of the tonsil. AB - Cells that are infected with HIV-1 were visualized at the mucosal surface of the nasopharyngeal and palatine tonsils in 14 specimens from patients with CD4+ T cell counts of 200 to 900/microliter and 2- to 10-year histories of HIV-1 infection. Most of the cells with intracellular HIV-1 protein were small but multinucleated. The majority of these syncytia could be double labeled for HIV-1 RNA and a dendritic cell marker S100. In the palatine tonsil, the infected cells were not found in the stratified squamous epithelium that is adjacent to the pharynx. Instead, the S100+ infected syncytia were localized to the surface of tonsil invaginations or crypts. This mucosa, termed lymphoepithelium, contains antigen-transporting M cells that lie above regions where S100+ dendritic cells are juxtaposed with CD4+ lymphocytes. Likewise, infected cells were found in lymphoepithelium and not respiratory epithelium of nasopharyngeal tonsils or adenoids. We propose that lymphoepithelia, the histological term that describes the specialized regions where antigens access mucosa-associated lymphoid tissue, are sites where HIV-1 replication can be enhanced in syncytia derived from dendritic cells. PMID- 9212736 TI - Human mucosal addressin cell adhesion molecule-1 is preferentially expressed in intestinal tract and associated lymphoid tissue. AB - Lymphocyte homing to normal tissues and recruitment to inflammatory tissue sites are controlled, in part, by the selective expression of chemokines, pro inflammatory cytokines and mediators, and various adhesion proteins and molecules. In the mouse, mucosal addressin cell adhesion molecule-1 (MAdCAM-1) is selectively expressed on endothelium of high endothelial venules in gut and gut associated lymphoid tissue. By interaction with its integrin ligand, alpha 4 beta 7, lymphocytes presumed to be involved in mucosal immunity are selectively recruited to these intestinal sites. After generating monoclonal antibodies against a murine cell line expressing recombinant human MAdCAM-1, we qualitatively and semiquantitatively assessed MAdCAM-1 expression in human tissue sections from various normal and inflammatory disorders. We found that human MAdCAM-1, as in the mouse, is expressed in a tissue-selective manner. In normal tissues, MAdCAM-1 is constitutively expressed to endothelium of venules of intestinal lamina propria. Interestingly, using computer-assisted morphometric analysis, the proportion of venular endothelium within lamina propria that expresses MAdCAM-1 is increased, compared with normal tissues, at inflammatory foci associated with ulcerative colitis and Crohn's disease. Moreover, for the most part, MAdCAM-1 is not detected in the majority of normal or inflamed extra intestinal tissues, including those with mucosal surfaces. These results are consistent with a role, as originally defined in the mouse, for human MAdCAM-1 in the localization of alpha 4 beta 7+ lymphocytes in the gastrointestinal tract and associated lymphoid tissue. As such, the pathway defined by MAdCAM-1/alpha 4 beta 7 may be a relevant tissue-specific therapeutic target for the modulation of inflammatory bowel disease activity. PMID- 9212737 TI - Activation of naive xenogeneic but not allogeneic endothelial cells by human naive neutrophils: a potential occult barrier to xenotransplantation. AB - Here we demonstrate that human neutrophils, the predominant circulating leukocytes in intimate contact with endothelial cells lining the vasculature, directly recognize xenogeneic endothelium independently of xenoreactive natural antibody and complement. A rapid and calcium-dependent activation of native (unstimulated) xenogenic endothelial cells by human neutrophils leads to 1) translocation of P-selectin from the Wiebel-Palade bodies to the surface of xenogeneic endothelial cells, 2) increased synthesis and expression of vascular cell adhesion molecule-1 on the xenogeneic endothelial cells, and 3) enhanced killing of the xenogeneic endothelium by natural killer cells. Our data directly implicate naive neutrophils as major early participants in xenograft recognition and endothelial activation independent of xenoreactive natural antibodies and complement. PMID- 9212738 TI - Enhanced lymphocyte longevity and absence of proliferation and lymphocyte apoptosis in Quilty effects of human heart allografts. AB - "Quilty effect" (QE) is a common and problematic observation in endomyocardial biopsy specimens from patients after cardiac transplantation. The origin, fate, and significance of QE cellular elements are unknown. Twenty-six paraffin embedded endomyocardial biopsy specimens with QE (five QE As and twenty-one QE Bs) from twenty-two cardiac allografts were studied by immunohistochemistry for expression of Bcl-2, Fas antigen, proliferating cell nuclear antigen (PCNA), perforin, T cells (UCHL-1), macrophages (CD68), and apoptosis by in situ terminal deoxyribonucleotide transferase (TdT)-mediated dUTP nick end labeling (TUNEL). Approximately 50% of the lymphocytes present, mainly in the deeper region of 20 of 21 QE Bs and all 5 QE As, expressed Bcl-2 in a pseudo-nodular pattern surrounding high endothelial venules. Fas expression was detected in lymphocytes in 20 of 21 QE Bs and 5 QE As in a similar pattern to Bcl-2. However, endothelial cells and macrophages were Bcl-2 negative, whereas both cell types were Fas positive. Perforin was negative in nearly all lymphocytes. TUNEL staining revealed that lymphocytes in QEs did not undergo apoptosis; however, TUNEL positivity was observed in approximately 70% of endothelial cells and macrophages and certain adjacent cardiac myocytes in 20 of 21 QE Bs and 5 QE As. One large QE B with a germinal center was noted. Germinal center cells expressed PCNA intensely but were negative for Bcl-2, Fas, and TUNEL. Cells surrounding the germinal center expressed abundant Bcl-2. The following conclusions were drawn. 1) Apoptosis does not occur in lymphocytes in QE where enhanced Bcl-2 (apoptosis inhibitor) and Fas antigen (apoptosis inducer) are expressed. 2) PCNA negativity indicates that QE lymphocytes may not proliferate, and perforin negativity indicates that they may not exhibit perforin-based cytotoxicity. We propose that there may be a relationship between the longevity of lymphocytes in QE and the absence of apoptosis. PMID- 9212739 TI - Systemic injection of products of activated neutrophils and H2O2 in myeloperoxidase-immunized rats leads to necrotizing vasculitis in the lungs and gut. AB - The strong association of anti-neutrophil cytoplasmic antibodies with various forms of systemic vasculitis suggests a role for these autoantibodies in the pathophysiology of systemic vasculitis. In the present study, we tested the hypothesis that release of neutrophil lysosomal enzymes in the presence of an anti-myeloperoxidase (anti-MPO) immune response may underlie the development of systemic vasculitis. Brown Norway rats were immunized with MPO in complete Freund's adjuvant or complete Freund's adjuvant alone. Two weeks after immunization, rats bad developed antibodies to human and rat MPO as measured by enzyme-linked immunosorbent assay. Next, rats were intravenously infused with 400 micrograms of a human neutrophil lysosomal extract containing 200 micrograms of MPO followed by 0.5 ml of a 1 mmol/L solution of H2O2 through a cannula inserted into the right jugular vein. Rats were sacrificed at 4 hours, 24 hours, 7 days, or 14 days, and several organs (lungs, heart, liver, spleen, gut, and kidneys) were examined for vasculitic lesions and inflammatory cell infiltrates. Macroscopically, patchy hemorrhagic spots were observed in the lungs and gut of MPO-immunized rats at days 7 and 14 after systemic infection of the neutrophil lysosomal extract and H2O2. Such changes were not observed at earlier time points or in control immunized rats. Histologically, the lungs of MPO-immunized rats sacrificed at days 7 and 14 showed patchy inflammatory cell infiltrates associated with vasculitis, granuloma formation, giant cells, and foci of hemorrhage. At 14 days, early signs of fibrosis were found with deposition of collagen and proliferation of fibroblasts. Furthermore, a prominent leukocytoclastic vasculitis was found in the small intestine of these rats characterized by fibrinoid necrosis and an extensive neutrophilic infiltrate. No inflammatory changes were found in the other organs studied (heart, liver, spleen, and kidneys). Control immunized rats, sacrificed at days 7 and 14 showed only some small foci of inflammatory infiltrates in the lungs whereas no inflammatory changes were found in the gastrointestinal tract. These studies show that release of products from activated neutrophils in the presence of anti-MPO autoantibodies may be relevant to the pathogenesis of anti-MPO-associated vasculitides. PMID- 9212740 TI - Role of interleukin-1 in mesangial cell proliferation and matrix deposition in experimental mesangioproliferative nephritis. AB - We examined the functional role of interleukin (IL)-1 in mesangial cell proliferation during rat anti-Thy-1 nephritis by blocking its action with IL-1 receptor antagonist (IL-1ra). Anti-Thy-1 nephritis was induced by intravenous injection of 5 mg/kg OX-7 IgG (day 0) into inbred Wistar rats. Groups of animals (n = 9) were implanted with a micro-osmotic pump on day -1, which delivered 25 micrograms/hour human recombinant IL-1ra or saline continuously until the rats were killed at day 6, the peak of mesangial cell proliferation. Immunostaining showed that IL-1 was expressed by mesangial cells during disease. IL-1ra treatment did not affect the mild, but significant, proteinuria seen after OX-7 injection. Compared with saline treatment, IL-1ra treatment reduced mesangial cell proliferation (decreases 24% P < 0.05), glomerular hypercellularity (decreases 29%; P < 0.05), and glomerular macrophage accumulation (decreases 20%; P < 0.05). However, IL-1ra treatment had no effect on glomerular IL-1 beta mRNA expression and caused only a small reduction in the high levels of glomerular expression of platelet-derived growth factor-beta protein (decreases 6%; P < 0.05). IL-1ra caused a modest reduction in the marked up-regulation of glomerular transforming growth factor-beta 1 mRNA expression on day 6 (decreases 26%; P < 0.05), although urinary excretion of this factor was unaffected. Interestingly, IL-1ra treatment had relatively little effect upon glomerular deposition of laminin, fibronectin, and collagen type IV seen in this acute disease. In conclusion, this study has 1) demonstrated that IL-1 is expressed by mesangial cells in vivo, 2) demonstrated that IL-1 is a mesangial cell growth factor in experimental mesangioproliferative nephritis, and 3) suggests that IL-1 has little or no fibrogenic activity in mesangial matrix deposition. PMID- 9212741 TI - Cyclin-dependent kinase inhibitor p27KIP1 in lymphoid tissue: p27KIP1 expression is inversely proportional to the proliferative index. AB - Cell cycle progression is regulated by the combined action of cyclins, cyclin dependent kinases (CDKs), and CDK inhibitors (CDKIs). p27KIP1, which has a high degree of similarity with p21WAF1, is a general CDKI thought to be involved in G1 arrest in response to agents that inhibit cell cycle progression. The aims of this study were 1) to establish the pattern of expression of p27KIP1 protein in nontumor lymphoid tissue, 2) to determine whether p27KIP1 is involved in lymphomagenesis, and 3) to address the possible relationship between p27KIP1 and p21WAF1 expression in reactive and tumor lymphoid tissue. p27KIP1 protein was found to be mainly present in quiescent lymphocytes in reactive lymphoid tissue as well as in peripheral blood lymphocytes, with an inverse expression for p27KIP1 and Ki-67 proteins. The same p27KIP1 expression pattern was observed in lymphomas, independently of histological type; small resting cells were p27KIP1 positive, and large proliferating cells were p27KIP1 negative. Therefore, tumors with a low proliferative index were mostly positive, whereas tumors characterized by a higher growth fraction bad low p27KIP1 protein levels. An unexpected finding was the existence of a group of six cases of high-grade lymphomas (three diffuse large B-cell lymphomas and three Burkitt's lymphomas) with homogeneously strong staining for p27KIP1 protein. All 6 of these cases belong to a group of 28 cases characterized by blockage of the p53 tumor suppressor pathway, as determined by genetic (p53 mutation) or immunophenotypic studies (p53+/p21-). p27KIP1 expression was not seen in any case of aggressive non-Hodgkin's lymphoma with an intact p53 pathway. The results indicate that p27KIP1 is down-regulated in lymphomas with a high proliferative index, although it is highly expressed in high-grade lymphomas with defects in the p53 pathway. PMID- 9212742 TI - Amplification of CCND1 and expression of its protein product, cyclin D1, in ductal carcinoma in situ of the breast. AB - Ductal carcinoma in situ (DCIS) of the breast is a heterogeneous disease clinically and biologically. The few available studies of its natural history implicate DCIS as a non-obligate precursor for invasive carcinoma. We have used fluorescence in situ hybridization (FISH) to detect gene amplification of the cell cycle regulator gene CCND1 in 88 examples of formalin-fixed, paraffin embedded DCIS. Expression of its protein product cyclin D1 was detected by immunohistochemistry. CCND1 was amplified in 18% of DCIS cases. High grade DCIS was more likely to show amplification than low grade DCIS (32% versus 8%; P = 0.08). Gene amplification was associated with cyclin D1 protein expression (P = 0.001), although cyclin D1 was detected in cases that did not demonstrate gene amplification. Overall, cyclin D1 protein was detected in 50% of DCIS cases. Although only 2 of 23 (8%) cases of low grade DCIS had CCND1 amplification, over 50% (13/23) of these cases expressed cyclin D1 protein. Low grade DCIS had a higher mean percentage of nuclei expressing cyclin D1 than did intermediate or high grade DCIS (P = 0.007). Mechanisms other than gene amplification may be responsible for increased cyclin D1 protein in DCIS, especially in low grade DCIS. Identifying mechanisms that control cell cycle progression in DCIS may yield clues to its biological behavior. PMID- 9212743 TI - Lymph vessel expansion and function in the development of hepatic fibrosis and cirrhosis. AB - The process of lymph vessel expansion and function in the development of CCl4 induced hepatic fibrosis and cirrhosis was studied using intravital fluorescence microscopy of the rat liver. The unique aspect of our approach was the use of high molecular fluorescein-isothiocyanate-labeled dextran (MW, 150,000) as fluorescent marker, which allowed for simultaneous assessment of both 1) the macromolecular blood hepatocytic exchange from the sinusoidal microvasculature (extra-/intrasinusoidal gray level intensity at 1, 3, 5, and 10 minutes after intravenous injection) and 2) the hepatic lymph system. In animals exposed with CCl4 up to 4 weeks, macromolecular trans-sinusoidal exchange was found progressively delayed. This was strongly associated with lymph vessel expansion and function, as indicated by a continuous increase of lymph vessel density and area. Delay of macromolecular exchange and lymph vessel expansion was found not further enhanced at fibrotic and cirrhotic stages of 8- and 12-week CCl4-exposed livers. Linear regression analysis revealed a strong negative correlation between lymphatic network density development and macromolecular trans-sinusoidal exchange (r2 = 0.99; P < 0.01). Thus, our study provides for the first time direct evidence for the pivotal role of lymphatic function for macromolecular transport in case of deteriorated sinusoidal hepatocellular exchange capacity. PMID- 9212744 TI - Interleukin-6 reduces cartilage destruction during experimental arthritis. A study in interleukin-6-deficient mice. AB - Using interleukin (IL)-6-deficient (IL-6(0/0) mice or wild-type mice, we investigated the controversial role of IL-6 in joint inflammation and cartilage pathology during zymosan-induced arthritis (ZIA). Monoarticular arthritis was elicited by injection of zymosan into the right knee joint cavity. Production of IL-1, tumor necrosis factor (TNF), IL-6, and nitric oxide by the inflamed knee was assessed in washouts of joint capsule specimens. Plasma corticosterone was measured using a radioimmunoassay. Proteoglycan synthesis was assessed using [35S]sulfate incorporation into patellas ex vivo. Joint swelling was quantified by joint uptake of circulating 99mTechnetium pertechnetate. Histology was taken to evaluate cellular infiltration and cartilage damage. Zymosan caused a rapid increase in articular IL-1, IL-6, TNF, and NO levels. Except for IL-6, the released amounts and time course of these mediators were comparable in the IL-6 deficient mice and the wild-type mice. Elevated plasma corticosterone levels were measured during the first day of arthritis in both strains. At day 2 of ZIA, joint inflammation (joint swelling and cell exudate) in IL-6-deficient mice was comparable with that in the wild-type mice. The marked suppression of chondrocyte proteoglycan synthesis and proteoglycan degradation were on the average higher in the IL-6-deficient mice. Together this resulted in a more pronounced proteoglycan depletion in the IL-6-deficient mice as compared with the wild-type mice during the first week of arthritis. Injection of recombinant IL-6 into the joint cavity corrected the IL-6 deficiency and significantly reduced cartilage destruction. Inflammation was more chronic in the wild-type mice, and these mice also showed a higher prevalence for osteophyte formation. In ZIA, IL-6 plays a dual role in connective tissue pathology, reducing proteoglycan loss in the acute phase and enhancing osteophyte formation in the chronic phase. The latter could be related to the more severe joint inflammation as seen in the normal (IL-6-producing) animals during the chronic phase of arthritis. PMID- 9212745 TI - Regulatory effects of interleukin-6 in immunoglobulin G immune-complex-induced lung injury. AB - Interleukin-6 (IL-6) is a cytokine produced in response to a variety of inflammatory stimuli. Although IL-6 is often observed in increased amounts in acute respiratory distress syndrome, its role in the development of lung injury is unclear. The role of IL-6 was studied in the rat model of lung injury induced by the intra-alveolar deposition of IgG immune complexes. IL-6 induction, as determined by Northern blot analysis and bioactivity, was found as a function of time during the course of development of injury. Recombinant IL-6 instilled intratracheally at commencement of injury led to substantial reductions in lung vascular permeability, neutrophil accumulation, and levels of tumor necrosis factor (TNF)-alpha and macrophage inflammatory protein (MIP)-2 in bronchoalveolar lavage fluids. Conversely, blocking of intrinsic IL-6 by a neutralizing antibody resulted in increases in lung vascular permeability, neutrophil content, and TNF alpha levels in bronchoalveolar lavage fluids. Rat alveolar macrophages stimulated in vitro with lipopolysaccharide in the presence of IL-6 showed a significant reduction in TNF-alpha expression. Together, these findings suggest that IL-6 acts as an intrinsic regulator of lung inflammatory injury after deposition of IgG immune complexes and that the protective effects of exogenously administered IL-6 may be in part linked to suppressed TNF-alpha production. PMID- 9212746 TI - Differential up-regulation of circulating soluble and endothelial cell intercellular adhesion molecule-1 in mice. AB - Although circulating levels of soluble intercellular adhesion molecule-1 (sICAM 1) are frequently used as an indicator of the severity of different immune, inflammatory, or neoplastic diseases, little is known about the factors that govern plasma sICAM-1 concentration and its relationship to the membranous form of ICAM-1 (mICAM-1) expressed on vascular endothelial cells. Plasma sICAM-1 concentration (measured by enzyme-linked immunosorbent assay) and mICAM-1 expression (measured using the dual radiolabeled monoclonal antibody technique) in different vascular beds (eg, lung, small intestine, and spleen) were monitored in wild-type (C57BL) and ICAM-1-deficient mice, before and after administration of tumor necrosis factor (TNF)-alpha. In wild-type mice, TNF-alpha elicited time dependent increases in lung and intestine mICAM-1 (plateau achieved at 12 hours), with a corresponding increase in plasma sICAM-1 (peaked at 5 hours and then declined). The initial increases in mICAM-1 and pulmonary leukocyte sequestration (measured as lung myeloperoxidase activity) induced by TNF-alpha preceded any detectable elevation in sICAM-1. In ICAM-1-deficient mice, plasma sICAM-1 was reduced by approximately 70%, with > 95% reductions of mICAM-1 in lung and intestine, and > 75% reduction in splenic accumulation of anti-ICAM-1 antibody. Although TNF-alpha doubled plasma sICAM-1 in ICAM-1-deficient mice, mICAM-1 was unaffected in all tissues. Either splenectomy or pretreatment with cycloheximide resulted in an attenuated TNF-induced increase in sICAM-1, without affecting mICAM-1 expression. These findings indicate that plasma sICAM-1 concentration does not accurately reflect the level of ICAM-1 expression on endothelial cells in different vascular beds. PMID- 9212747 TI - Reactive oxygen species cause direct damage of Engelbreth-Holm-Swarm matrix. AB - Reactive oxygen species (ROS) are produced and released into the extracellular spaces in numerous diseases and contribute to development and progression, for example, of inflammatory diseases, proteinuria, and tumor invasion. However, little is known about ROS-induced chemical changes of interstitial matrix proteins and their consequences for the integrity of the matrix meshwork. As basement membranes and other matrices are highly cross-linked and complex, the relatively simple matrix produced by Engelbreth-Holm-Swarm (EHS) sarcoma, and proteins isolated therefrom, were incubated in vitro with defined concentrations of ROS that were generated by the Fenton or xanthine oxidase/xanthine reactions. This resulted in two counter-current effects. Although up to approximately 15% of the EHS matrix proteins were released into the supernatant in a ROS dose-response relationship, the residual insoluble matrix was partially cross-linked by ROS. Matrix proteins released into the supernatants were examined by rotary shadowing, quantitative sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblotting, and fluorospectrometry for loss of tryptophans and formation of bityrosine residues. At relatively low ROS concentrations, selective liberation of morphologically intact laminin/entactin was found that, however, failed to reassociate and showed oxidative damage of its tryptophan residues. At higher ROS concentrations, laminin and entactin were progressively disintegrated, partially fragmented, and eventually completely degraded. At this point oligomers of type IV collagen predominated in the supernatant, and proteoglycans were not encountered at any concentration of ROS. Similar gradual molecular changes were also obtained when fractions of isolated soluble EHS matrix proteins were incubated with graded concentrations of ROS. In these experiments, the formation of covalently linked oligomers and aggregates paralleled the ROS-dependent formation of cross-linking bityrosine groups. ROS scavengers pinpointed to the hydroxyl radical as the most damaging radical species. Protease inhibitor experiments suggested that degradation of matrix proteins was caused primarily by the direct action of ROS and not by proteolysis by potentially contaminating proteases. Collectively, these results provide evidence that EHS matrix proteins show differential sensitivity to ROS-induced damage in a reproducible, sequential pattern, in the order entactin > laminin > type IV collagen, and that ROS cause partial dissociation and cross-linking of the EHS matrix. PMID- 9212748 TI - Inhibition of colon carcinoma cell lung colony formation by a soluble form of E selectin. AB - During metastasis, tumor cells adhere to vascular endothelia. E-selectin is an adhesive protein expressed by cytokine-activated endothelium that can support adhesion of colon cancer cells through the recognition of specific carbohydrate ligands. Using a series of colon carcinoma cell lines that displayed E-selectin adhesiveness and an increased metastatic capacity in cytokine-treated mice, we examined possible inhibition of cytokine-dependent experimental lung metastasis by a soluble form of E-selectin, the recombinant fusion protein E-selectin immunoglobulin. We found that E-selectin-immunoglobulin bound to the surfaces of HT-29 colon carcinoma cells and blocked the formation of cytokine-inducible experimental lung metastases; control L-selectin-immunoglobulin also bound to HT 29 cells but had no effect on tumor cell lung colonization. E-selectin immunoglobulin was found to interfere with E-selectin-dependent adhesion of HT-29 cells to activated vascular endothelium and to block the retention of these cells in the lung, a process that implies tumor cell adhesive interactions with the host vasculature. Our results demonstrate that E-selectin-immunoglobulin inhibits adhesion and formation of lung metastases by colon carcinoma cells and suggest that impairment of tumor cell-endothelium adhesion might represent a therapeutic approach to the metastatic diffusion of tumors. PMID- 9212749 TI - Expression of membrane-type 1 matrix metalloproteinase and activation of progelatinase A in human osteoarthritic cartilage. AB - Matrix metalloproteinases (MMPs) are expressed in osteoarthritic (OA) cartilage and are thought to be involved in the degradation of cartilage extracellular matrix (ECM). Among these proteinases, MMP-2 (gelatinase A) demonstrates a wide range of substrate specificity against the ECM present in cartilage. Although MMP 2 expression increases in OA cartilage, the activation mechanism of the corresponding zymogen (pro-MMP-2) in cartilage is unknown. In this study, we examined the expression pattern of membrane-type 1 MMP (MT1-MMP) in human OA articular cartilage and its correlation with the activation of pro-MMP-2. Immunohistochemical studies demonstrate that MT1-MMP localizes to the chondrocytes in the superficial and transitional zones in all of the samples examined directly correlating with cartilage degradation. Reverse transcription polymerase chain reaction confirmed the predominant expression of MT1-MMP mRNA in the OA cartilage. In situ hybridization revealed the site of expression of MT1 MMP in OA cartilage to be the chondrocytes. Through gelatin zymography and a sandwich enzyme immunoassay it was demonstrated that OA cartilage explants secrete significantly higher levels of pro-MMP-2 than normal samples. Pro-MMP-2 activation was enhanced in the OA cartilage samples and correlated with MT1-MMP expression in the cartilage. Plasma membranes prepared from cultured chondrocytes with MT1-MMP expression and those directly isolated from OA cartilage could activate pro-MMP-2. MT1-MMP gene expression in cultured chondrocytes was induced by treatment with interleukin-1 alpha and/or tumor necrosis factor-alpha. These data suggest that cytokine-induced MT1-MMP in the chondrocytes may play a key role in the activation of pro-MMP-2 in the OA articular cartilage, leading to cartilage destruction through ECM degradation. PMID- 9212750 TI - Post-infarction left ventricular remodeling induces changes in creatine kinase mRNA and protein subunit levels in porcine myocardium. AB - Energy metabolism is altered in post-infarction remodeled pig myocardium. To understand the basis of this abnormality, we examined the pattern of creatine kinase (CK) gene expression and the relative content of CK protein subunits in pig hearts with proximal left circumflex coronary artery ligation. At 2 months after infarct, both Northern and Western blot analyses were performed on left ventricular myocardium remote from the infarct zone in ligation animals (n = 8). Results were compared with data from the left ventricular myocardium from similar sized normal (control) pigs (n = 7). Steady-state levels of mitochondrial CK mRNA decreased 46% in left ventricular remodeled (LVR) heart samples (93.40 +/- 18.60 arbitrary units) compared with controls (172.85 +/- 37.20 arbitrary units), whereas CK-M subunit mRNA levels remained unchanged between the control and LVR groups (319.50 +/- 35.25 and 352.50 +/- 62.18 arbitrary units, respectively). The mean control group CK-M protein subunits (2.04 +/- 0.31 arbitrary units) decreased 53% (P < 0.05) compared with the LVR group (0.95 +/- 0.25 arbitrary units). Similarly, the mean control group (n = 4) mitochondrial CK protein subunits (1.12 +/- 0.04 arbitrary units) decreased 30% (P < 0.05) compared with the LVR group (n = 4; 0.79 +/- 0.06 arbitrary units). Mean CK-B protein subunits in LVR pig hearts (0.84 +/- 0.23 arbitrary units) increased 77% compared with control (0.48 +/- 0.05 arbitrary units). The total CK activity did not change significantly between control hearts at 164 +/- 11 IU/mg and LVR at 212 +/- 32 IU/mg. We suggest that these alterations of the CK system represent the bioenergetic phenotype of LVR myocardium at the molecular level. The CK system response may ultimately prove inadequate in meeting the abnormal energy requirements of remodeled heart and, therefore, may contribute to the transition toward failure. PMID- 9212751 TI - Carboxyl-terminal fragments of beta-amyloid precursor protein bind to microtubules and the associated protein tau. AB - Alzheimer's disease is a neurodegenerative disorder characterized by protein depositions in intracellular and extracellular spaces in the brain. The intraneuronal deposits are formed by neurofibrillary tangles composed mainly of abnormally phosphorylated tau, a microtubule-associated protein, whereas the major constituent of the amyloid deposited extracellularly in the brain parenchyma and vessel walls is amyloid beta-protein (A beta). The proteolytic processing of the beta-amyloid precursor protein (beta PP) results in the generation of a complex set of carboxyl-terminal peptides that contain A beta. In this study, we have used fusion proteins containing carboxyl-terminal fragments of beta PP to investigate the association of beta PP with cellular components. We demonstrate that specific domains within the carboxyl end of beta PP contain binding sites for cytoskeletal components; one, within residues 1 to 28 of A beta, binds directly to tubulin, and the second one, within sequences carboxyl terminal to A beta, binds tau and tubulin. We propose that the two neuropathological hallmarks of Alzheimer's disease, A beta deposition and neurofibrillary tangles, represent the residual of a disrupted beta PP-tubulin tau complex. PMID- 9212752 TI - Helicobacter mustelae-associated gastric MALT lymphoma in ferrets. AB - Gastric lymphoma resembling gastric mucosa-associated lymphoid tissue (MALT) lymphoma linked with Helicobacter pylori infection in humans was observed in ferrets infected with H. mustelae. Four ferrets with ante- or postmortem evidence of primary gastric lymphoma were described. Lymphoma was diagnosed in the wall of the lesser curvature of the pyloric antrum, corresponding to the predominant focus of H. mustelae induced gastritis in ferrets. Two ferrets had low-grade small-cell lymphoma and two ferrets had high-grade large-cell lymphoma. Gastric lymphomas demonstrated characteristic lymphoepithelial lesions, and the lymphoid cells were IgG+ in all ferrets. Lymphoma was confirmed by light chain restriction, which contrasted with the 1.2:1 kappa lambda ratio observed in H. mustelae-associated chronic gastritis. H. mustelae infection in ferrets has been used as a model for gastritis, ulcerogenesis, and carcinogenesis. The ferret may provide an attractive model to study pathogenesis and treatment of gastric MALT lymphoma in humans. PMID- 9212755 TI - Immunohistochemical localization of DNA topoisomerase II in human gastric disorders. PMID- 9212753 TI - Transgenic mice with increased expression of vascular endothelial growth factor in the retina: a new model of intraretinal and subretinal neovascularization. AB - Vascular endothelial growth factor (VEGF) has been implicated in retinal neovascularization (NV), but it has been difficult to produce retinal NV with exogenous VEGF. We investigated the effect of increased VEGF expression in the retina using tissue-specific, gain-of-function transgenic mice in which the bovine rhodopsin promoter is coupled to the gene for human VEGF. Three founder mice were obtained and used to generate transgenic lines. One of the lines shows increased expression of VEGF in the retina by reverse transcription coupled to polymerase chain reaction and Northern blots, and the VEGF is localized to photoreceptors by immunohistochemistry. These mice demonstrate new vessels originating from the deep capillary bed of the retina that extend beneath the photoreceptor layer into the subretinal space where they form clumps of blood vessels surrounded by proliferated retinal pigmented epithelial cells. The appearance is similar to subretinal NV seen in some patients, except that the blood vessels originate from the retinal vasculature rather than the choroidal vasculature. One of the other two lines of mice did not show increased expression of VEGF and did not have NV; the other line showed retinal degeneration. This study demonstrates that over-expression of VEGF in the retina is sufficient to cause intraretinal and subretinal NV and provides a valuable new animal model. PMID- 9212754 TI - Harlequin ichthyosis (ichq): a juvenile lethal mouse mutation with ichthyosiform dermatitis. AB - The harlequin ichthyosis (ichq) mouse mutation arose spontaneously in 1989 in a colony of BALB/cJ mice at The Jackson Laboratory. Affected mice developed thick skin due to formation of compact, orthokeratotic scales that fractured over articular surfaces, secondary to bending. Harlequin ichthyosis mice on the inbred BALB/cJ background died between 9 and 12 days of age. Onset of the clinical phenotype corresponded with emergence of hair fibers from follicles at 5 days of age. There was marked proliferation of the root sheaths of anagen hair follicles, limited to the region within the dermis. Sebaceous glands were present but small compared with those of littermate controls. Emerging hair fibers were surrounded by a thick, compact sheath of cornified cells. Mutant skin contained large mitochondria with lamellar-shaped, electron-dense structures at the ultrastructural level. Keratohyalin granules were smaller and less pleomorphic than those in control mice. Lamellar bodies were not evident in either mutant or littermate control mice. Using a panel of antibodies to evaluate changes in keratinocyte differentiation, mouse-specific keratin 6 was overexpressed in the suprabasilar, hyperplastic epidermis. Loricrin expression, within the cytoplasm of cells in the stratum granulosum, decreased rapidly postmortem, unlike that in normal mice where it was stable for over 24 hours postmortem. Filaggrin expression, within granules of cells in the stratum granulosum, was prominent, corresponding to hypergranulosis evident by light microscopy in mutant mouse skin. Skin grafts from harlequin ichthyosis mice grafted onto immunodeficient nude mice maintained the phenotype for the 10-week observation period. The mutant gene locus mapped to the proximal end of mouse chromosome 19 and is inherited as a fully penetrant autosomal recessive gene. The harlequin ichthyosis mouse mutation is very similar to human type 2 harlequin ichthyosis for which it may be a good model. PMID- 9212756 TI - Intercellular adhesion molecule (ICAM)-3 expression on endothelial cells. PMID- 9212757 TI - [Mibefradil opens up a new class of calcium antagonists. Glasgow, 23 June 1996]. PMID- 9212758 TI - The 80th anniversary of the Mayo Department of Ophthalmology. PMID- 9212759 TI - Photodynamic therapy for early stage squamous cell carcinoma of the lung. AB - OBJECTIVE: To study the effectiveness of photodynamic therapy (PDT) as a therapeutic strategy in roentgenographically occult squamous cell carcinoma of the lung. MATERIAL AND METHODS: A carefully selected group of 21 patients (with 23 cancers) who had early stage squamous cell carcinoma of the lung and were eligible for surgical treatment were offered PDT as an alternative to resection. Patients underwent close follow-up with bronchoscopic surveillance and were offered resection if cancer persisted after no more than two sessions of PDT. RESULTS: A complete response was identified in 15 patients (16 cancers) after an initial PDT session. A complete response that lasted longer than 12 months was noted in 11 patients (52%). After PDT, the minimal follow-up period was 24 months. A subsequent primary lung cancer developed in 5 of the 21 patients (24%). Ten patients ultimately had surgical treatment, in 3 (30%) of whom N1 disease was identified at the time of resection. Two patients refused a surgical procedure and received alternative therapy. Therefore, nine patients (43%) were spared an operation (95% confidence interval, 21.8 to 66.6%). The mean duration of follow up for these nine patients was 68 months (range, 24 to 116). CONCLUSION: On the basis of this investigation, we can conclude with 95% confidence that at least 22% of patients with early stage squamous cell lung cancer who are candidates for PDT can be spared surgical resection. PMID- 9212760 TI - Eosinophilic myocarditis manifesting as myocardial infarction: early diagnosis and successful treatment. AB - OBJECTIVE: To report a case of eosinophilic myocarditis with remarkable initial clinical manifestations and outcome. MATERIAL AND METHODS: A 67-year-old woman with hypertension and a history of asthma and drug hypersensitivity was referred to our institution with a diagnosis of acute myocardial infarction on the basis of severe chest pain, ST elevation on an electrocardiogram, and a slight increase in cardiac enzymes. Further diagnostic studies were performed. RESULTS: Echocardiography disclosed left ventricular dysfunction in conjunction with apical asynergy, thinning, and thrombus. The eosinophil count in the peripheral blood was increased only slightly. Coronary angiography showed normal arteries and prompted the performance of endomyocardial biopsy, which revealed active eosinophilic myocarditis. After corticosteroid therapy, global and regional left ventricular function returned to normal. CONCLUSION: This unusual clinical picture and outcome demonstrate that eosinophilic myocarditis may simulate acute myocardial infarction and should be considered in patients with a history of allergies or acute left ventricular dysfunction, even in the absence of pronounced eosinophilia in the peripheral blood. With appropriate medical therapy, recovery for these patients can be complete. PMID- 9212761 TI - A new autosomal dominant disorder of pyogenic sterile arthritis, pyoderma gangrenosum, and acne: PAPA syndrome. AB - OBJECTIVE: To describe a multigenerational family with transmission of an autosomal dominant disorder characterized by pyogenic arthritis, pyoderma gangrenosum, and severe cystic acne. MATERIAL AND METHODS: We present a detailed case report of a 39-year-old man with arthritic changes in several joints, pyoderma gangrenosum, and cystic acne. Several relatives from three generations of his family underwent clinical and genetic investigations. The findings in this kindred are reported. RESULTS: Ten affected family members in three generations manifested variable expression of a pauciarticular, nonaxial, destructive, corticosteroid-responsive arthritis that began in childhood; pyoderma gangrenosum; and severe cystic acne in adolescence and beyond. Other less commonly associated features included adult-onset insulin-dependent diabetes mellitus, proteinuria, abscess formation at the site of parenteral injections, and cytopenias attributable to sulfonamide medications. Laboratory evaluation was nondiagnostic. Genetic studies excluded linkage to the major histocompatibility locus. CONCLUSION: The acronym of PAPA syndrome (pyogenic sterile arthritis, pyoderma gangrenosum, and acne) is suggested for this newly recognized pleiotropic autosomal dominant disorder. The nature of the genetic alteration in PAPA syndrome is unknown. PMID- 9212762 TI - Dexpanthenol enemas in ulcerative colitis: a pilot study. AB - OBJECTIVE: To test the hypothesis that topical administration of pantothenic acid, a precursor of coenzyme A, might result in increased tissue levels of coenzyme A, improvement of fatty acid oxidation, and amelioration of ulcerative colitis. MATERIAL AND METHODS: In an open-label pilot study, three patients with active left-sided ulcerative colitis received nightly enemas that contained 1,000 mg of dexpanthenol for 4 weeks. Before and after the study, patients submitted stool specimens for short-chain fatty acid analysis and urine collections for measurement of pantothenic acid and dicarboxylic acids; they also underwent flexible sigmoidoscopy for procurement of biopsy specimens for histologic examination and measurement of colonic coenzyme A activity. A clinical disease activity index and histologic disease activity index were used to assess response. RESULTS: Despite increases in urinary pantothenic acid, no significant changes were found in colonic tissue coenzyme A concentrations, fecal short-chain fatty acid concentrations, or urinary dicarboxylic acid concentrations. Moreover, no significant changes in clinical or histologic disease activity were noted. Although stool frequency and rectal bleeding remained unchanged, all patients noted increased abdominal cramping, and one patient had an increased extent of disease. CONCLUSION: Topically administered dexpanthenol seems to be absorbed, but at the dose used in this study, it did not influence concentrations of colonic coenzyme A activity, fecal short-chain fatty acids, or clinical response in patients with active left-sided ulcerative colitis. PMID- 9212763 TI - Prophylactic intravenous administration of caffeine and recovery after ambulatory surgical procedures. AB - OBJECTIVE: To determine whether prophylactic intravenous administration of caffeine, to daily caffeine users, decreases the frequency of postoperative headache and shortens recovery time. DESIGN: The study was a prospective, randomized, double-blind investigation with predetermined sample size and statistical power. MATERIAL AND METHODS: After Mayo Institutional Review Board approval and informed consent were obtained, 300 adult ambulatory surgical patients were enrolled in this study, which included randomization to receive either placebo or caffeine (200 mg intravenously) in the postanesthesia care unit. While recuperating, patients were allowed their choice of postoperative beverages. Before dismissal, patients completed a questionnaire providing details about intake of caffeine and tobacco, history of headache, and demographic data. Patients were considered "at risk" for symptoms of caffeine withdrawal if they did not drink a caffeinated beverage after the surgical procedure. RESULTS: Completed questionnaires were obtained from 234 patients. Patients at risk for symptoms of caffeine withdrawal were less likely to have a postoperative headache if they received caffeine intravenously rather than placebo-10% versus 23% (P < 0.05). Time until recovery was not significantly different between caffeine and placebo study groups. CONCLUSION: We conclude that prophylactic intravenous administration of caffeine was beneficial for those patients at risk for symptoms of caffeine withdrawal. For patients who consume caffeinated beverages on a daily basis, we recommend prophylactic administration of caffeine on the day of an ambulatory surgical procedure and anesthesia. PMID- 9212764 TI - Management of postpolio syndrome. AB - Recent research has shed light on the pathogenesis of the postpolio syndrome and has helped explain its symptoms and the rationale for management. The aim of this article is to familiarize physicians with this syndrome. The history, acute infection, definition, and diagnosis are discussed, as well as the various symptoms and their management. People with postpolio syndrome can educate health professionals about this condition and can help others inflicted with this syndrome. Thus far, no cure is available. A correct diagnosis is important, and the physician must realize that severe comorbidities tend to afflict people with this syndrome. Numerous management options are available to help these people enjoy a high quality of life. PMID- 9212765 TI - Organizing diffuse alveolar damage associated with progressive systemic sclerosis. AB - Diffuse alveolar damage (DAD) is a relatively nonspecific pattern of acute lung injury that can be observed in a wide range of clinical circumstances. DAD has often been recognized in association with various connective tissue diseases; however, to our knowledge, it has not been previously reported in the setting of progressive systemic sclerosis. Herein we describe two patients with established diagnoses of progressive systemic sclerosis who had development of the acute respiratory distress syndrome. Open-lung biopsy specimens from both patients showed a histologic pattern of DAD with no identifiable cause other than their progressive systemic sclerosis. Our results suggest that DAD should be added to the list of pleuropulmonary complications of progressive systemic sclerosis. PMID- 9212766 TI - Azathioprine-induced lymphoma manifesting as fulminant hepatic failure. AB - Azathioprine and rheumatoid arthritis are known to be associated with an increased risk of the development of non-Hodgkin's lymphoma; however, the manifestation of fulminant hepatic failure is extremely uncommon in patients with non-Hodgkin's lymphoma. In this article, we describe a patient with rheumatoid arthritis who was taking azathioprine, in whom fulminant hepatic failure occurred because of massive lymphomatous infiltration of the liver. PMID- 9212767 TI - Rumination syndrome. AB - Rumination is a syndrome characterized by repetitive regurgitation of small amounts of food from the stomach. The food is then partially or completely rechewed, reswallowed, or expelled. This syndrome is relatively common in infants and mentally challenged persons, but it also occurs in adults with normal intelligence. The rumination syndrome is an underappreciated condition in adults who frequently receive a misdiagnosis of vomiting due to gastroparesis or gastroesophageal reflux. Difficulties in establishing the correct diagnosis may be caused by a lack of awareness of the condition among physicians. This syndrome must be considered in the differential diagnosis of a patient with regurgitation, vomiting (especially postprandial), and weight loss. Reassurance, explanations, and behavioral therapy are currently the mainstays of treatment in adults with normal intelligence who have the rumination syndrome. Appropriately controlled trials are needed to establish the best therapy. PMID- 9212769 TI - Ukrainian physicist contributes to the discovery of X-rays. PMID- 9212768 TI - Office management of ovarian cysts. AB - Ovarian cysts are detected in female patients of all ages. The patient's age, the size of the cyst, and the ultrasound appearance are helpful in determining which ovarian cysts necessitate observation and which necessitate surgical excision. The cancer antigen 125 level alone does not help to distinguish between benign and malignant ovarian cysts. The combination of benign findings on pelvic examination, a benign ultrasound appearance, and a cancer antigen 125 level within normal limits indicates a benign origin in practically all cases. PMID- 9212770 TI - Survey of physician leadership and management education. AB - Health-care organizations have recognized the need to prepare physicians for various leadership and management positions within their own institutions. In the past, those who desired further education had to search beyond the boundaries of their practice to fulfill this need. The demands of a dynamic and changing health care environment have created increased pressure on organizations to develop a larger cadre of physician leaders and managers among their staff and to accomplish this outcome in a cost-effective, efficient manner. This article examines the results from a survey of leading medical institutions on the existence of in-house leadership and management educational programming. It also documents the approaches used by the responding organizations and the content of their course work. Numerous institutions are accepting the challenge for increased physician expertise in leadership and management by developing their own in-house programs. Future directions for Mayo initiatives in succession planning will be obtained from this benchmark survey. PMID- 9212771 TI - Multiple sclerosis: insights into molecular pathogenesis and therapy. PMID- 9212772 TI - The controversy surrounding the pathogenesis of the multiple sclerosis lesion. AB - The main issues in multiple sclerosis research revolve around four fundamental questions. (1) What initiates the disease-that is, autoimmune T cells, a virus, or a toxin? (2) Is the inflammatory response primary to the development of demyelination, or is it a secondary response to injury? (3) Is the oligodendrocyte, the myelin-producing cell, the primary target? (4) How can myelin repair be promoted? This review focuses on the controversies revolving around these important questions. Although many investigators believe that T-cell receptors on CD4+ cells interact with myelin antigens to initiate an inflammatory cascade that leads to myelin destruction, others maintain that a viral agent may have a direct or indirect role in the pathogenesis of multiple sclerosis. The concept that the immune system contributes to the tissue destruction in multiple sclerosis is generally accepted; however, the debate about cause versus consequence of the pathologic process remains unresolved, as does the identification of the initial event or focus of the damage. Electron microscopic studies have disclosed evidence of remyelination (albeit often incomplete) in lesions of multiple sclerosis. Enhanced understanding of the factors limiting remyelination could help formulate strategies to promote repair. By innovative experimental design and application of available molecular techniques, the answers to these questions may provide insights on how to prevent or treat multiple sclerosis. PMID- 9212773 TI - 3-year-old boy with cystic fibrosis and abdominal pain. PMID- 9212774 TI - The yellow brick road to penicillin: a story of serendipity. AB - Approximately 14 years elapsed between Sir Alexander Fleming's discovery of penicillin (in 1928) and its full-scale production for therapeutic use (in 1942) in World War II. The following factors were responsible for the delay: a scientific explanation of Fleming's "phenomenon," classification of the fungus secreting the active substance, source of the mold, initial difficulty of other bacteriologists in reproducing Fleming's discovery, identifying the chemical makeup of penicillin, search for other penicillin-producing organisms to enhance production of penicillin, purification and crystallization of penicillin, experiments on animals (chiefly mice) to determine toxicity, hesitancy to administer the drug to humans, standardization of an effective dosage for humans, and search for equipment and financial resources to enhance full-scale production. The adjunctive role of serendipity (chance, happenstance, improbability, and luck) in overcoming these obstacles and in contributing to the successful, scientific conclusion of the penicillin project is an unusual story. PMID- 9212775 TI - Photodynamic therapy for early stage central type of lung cancer. PMID- 9212776 TI - Interrelationship between cellular calcium homeostasis and free radical generation in myocardial reperfusion injury. AB - This review describes the interrelationship between two important biological factors, intracellular calcium overloading and oxygen-derived free radicals, which play a crucial role in the pathogenesis of myocardial ischemic reperfusion injury. Free radicals are generated during the reperfusion of ischemic myocardium, and polyunsaturated fatty acids in the membrane phospholipids are the likely targets of the free radical attack. On the other hand, activation of phospholipases can provoke the breakdown of membrane phospholipids which results in the activation of arachidonate cascade leading to the generation of prostaglandins, and oxygen free radicals can be produced during the interconversion of the prostaglandins. In conclusion, it has been emphasized that the two seemingly different causative factors of reperfusion injury, intracellular calcium overloading and free radical generation are, in fact, highly interrelated. PMID- 9212777 TI - Reductive activation of doxorubicin by xanthine dehydrogenase from EMT6 mouse mammary carcinoma tumors. AB - The role of enzymes in the reductive activation of various chemotherapeutic agents is an area of considerable interest in studies to better understand the selective toxicities of these agents. Xanthine dehydrogenase (XDH) is an enzyme capable of reductive activation of chemotherapeutic agents. Previously, this enzyme has not been extensively studied because of difficulties in its isolation. We recently isolated this enzyme from EMT6 mouse mammary carcinoma cells and showed that this enzyme is capable of activating mitomycin C. In this study, we examined whether XDH could activate the clinically important antineoplastic agent, doxorubicin. Drug activation was determined under aerobic and hypoxic conditions and at various pHs in order to simulate the different environments found in solid tumors. The results of these studies show that XDH reacts with doxorubicin via a two-electron reduction. This reduction is different from the modified and more extensively studied form of the enzyme, xanthine oxidase (XO), which reacts with doxorubicin via a one-electron reduction. Under hypoxic conditions, the formation of large quantities of 7-deoxydoxorubicin aglycone, a deactivation product of doxorubicin metabolism, may serve to moderate doxorubicin's antineoplastic activity. Under aerobic conditions, however, XDH activation led to a greater rate of formation of oxygen radicals than XO thereby possibly potentiating doxorubicin's cytotoxicity to aerobic tumor cells. Kinetic constants were determined for doxorubicin activation by XDH. PMID- 9212778 TI - Disposition of butadiene epoxides in Sprague-Dawley rats. AB - 1,2-Epoxybutene (BMO) and diepoxybutane (BDE) are metabolic products of 1,3 butadiene in rodents. Both BMO and BDE are suspect in the development of tumors in rats and mice. To understand the distribution and elimination of these compounds in the absence of the rate-limiting production from butadiene, the pharmacokinetics of BMO and BDE in blood were determined in adult male Sprague Dawley rats following intravenous administration. All animals were dually cannulated in these studies. For the BMO studies, rats were dosed with 71, 143, or 286 mumol/kg BMO (n = 3 for each dose group). For the BDE studies, rats were dosed with 523 mumol/kg BDE (n = 3). All animals tolerated the BMO and BDE doses without grossly observable adverse effects. Blood was drawn at predetermined time points and extracted in methylene chloride. BDE and BMO concentrations were quantitated by gas chromatography or gas chromatography/mass spectrometry. The BMO distribution half-lives were short and ranged from 1.4 min at the lowest dose to 1.8 min at the highest dose. Volume of distribution at steady state ranged from 0.53 +/- 0.17 to 0.59 +/- 0.31 l/kg. Systemic clearances ranged from 67 +/- 17 to 114 +/- 20 ml/min per kg. The terminal elimination half-lives were also short and ranged from 5.7 to 8.5 min among the doses. The pharmacokinetic parameters after an i.v. dose of 523 mumol/kg BDE were a distribution half-life of 2.7 min, terminal elimination T1/2 of 14 min, volume of distribution at steady state of 0.73 +/- 0.06 l/kg, and systemic clearance of 76 +/- 8 ml/min per kg. These pharmacokinetic parameters demonstrate the similarity between disposition of the two epoxides in rats, that include a rapid distribution after i.v. administration into a small extravascular body compartment as well as a rapid elimination from blood. These pharmacokinetic data provide useful blood clearance information for assessing the critical physiological and biochemical determinants underlying the disposition of butadiene epoxides. PMID- 9212779 TI - Structural features associated with reactive metabolite formation in clozapine analogues. AB - Clozapine is associated with a high incidence of agranulocytosis. We had previously found that it is oxidized by granulocytes, or simply HOCl, to a reactive metabolite that irreversibly binds to the cells, and we proposed that this reactive metabolite is responsible for clozapine-induced agranulocytosis. The reactive metabolite appeared to be a nitrenium ion formed by chlorination of the nitrogen bridge between the two aromatic rings. If this is correct, analogs that contain this structural feature should also be oxidized to a reactive intermediate while those not possessing this feature would, at least, not form the same type of reactive intermediate and, therefore, may not induce agranulocytosis. We tested the first part of this hypothesis with three clozapine analogs that do contain a nitrogen bridge and three that do not. Consistent with the hypothesis, the three analogs that do contain the nitrogen bridge formed reactive intermediates that could be trapped with glutathione when oxidized by HOCl, myeloperoxidase or activated neutrophils. In contrast, we found no evidence of a reactive intermediate on oxidation of analogs that contained an oxygen or sulfur bridge rather than a nitrogen bridge. If such reactive metabolites are responsible for drug-induced agranulocytosis, it should be possible to use such a simple screening method to test drugs at an early stage in their development for the potential to induce agranulocytosis. PMID- 9212781 TI - MP8-dependent oxidative dehalogenation: evidence for the direct formation of 1,4 benzoquinone from 4-fluorophenol by a peroxidase-type of reaction pathway. AB - The present study shows that MP8 in the presence of H2O2 is able to catalyze the rupture of the stable carbon-fluorine bond of 4-fluorophenol, used as a model substrate for the oxidative dehalogenation reaction. 1,4-Benzoquinone was shown to be the primary reaction product. It is also demonstrated that there was significant [18O] incorporation into the product, 1,4-benzoquinone, from 18O labelled H2(18)O but not from H2(18)O2. This implies that water participates in the reaction mechanism, and acts as a source for the oxygen atom inserted into the product. It also suggests that the reaction is not a result of direct oxygen transfer from H2O2 through the heme catalyst to the product. Furthermore, ascorbic acid, known to efficiently block MP8-catalyzed peroxidase-type conversions, inhibits the MP8-dependent dehalogenation reaction, most likely because of its ability to reduce the phenoxy radical back to the parent substrate. This observation together with the above-mentioned incorporation of oxygen from the solvent into the benzoquinone product indicates that MP8 dehalogenates 4-fluorophenol and converts it to 1,4-benzoquinone in a peroxidase- and not a P-450-type of reaction mechanism. Overall, our results indicate that the oxidative dehalo genation of para-halogenated phenols, resulting in the formation of benzoquinones, is not specific only for cytochrome P-450 enzymes. Hemoproteins exhibiting peroxidase activity could also play a role in the metabolism of these xenobiotics, resulting in the formation of electrophilic reactive benzoquinone type metabolites. PMID- 9212780 TI - Effects of peroxisome proliferators and/or hypothyroidism on xenobiotic metabolizing enzymes in rat testis. AB - The objectives of the present work were to study the effects of certain peroxisome proliferators on xenobiotic-metabolizing enzyme activities in the testes of normal and hypothyroid rats, i.e. phenol sulfotransferases (pST), phenol UDP-glucuronosyl transferases (pUDPGT), glutathione transferases (GST), catalase, epoxide hydrolase (EH), glutathione peroxidase (GPX) and NAD(P)H quinone oxidoreductase (QR). Adult male rats (normal and hypothyroid) were treated for 10 days with clofibrate (0.5%), perfluorooctanoic acid (0.5%, PFOA), acetylsalisylic acid (1%, ASA) and di(2-ethylhexyl)phthalate (2%, DEHP) in their diet. The results show that treatment of normal rats with peroxisome proliferators dramatically affects the activities of xenobiotic-metabolizing enzymes (40-60% reduction). The highest effects are seen in catalase activity (50 60% with PFOA and ASA), pUDPGT (55% with PFOA), pST (55% with PFOA) and QR (50% with DEHP). These effects are not seen or are weaker after induction of hypothyroidism. Taken together, it is concluded that different classes of peroxisome proliferators have different effects on rat testicular xenobiotic metabolizing enzymes. PMID- 9212783 TI - Structural specificity requirements in the binding of beta lactam antibiotics to human serum albumin. AB - The binding of some cephalosporins of pharmacological interest, to human serum albumin was studied using ultrafiltration method. The identification of the binding sites in albumin was also performed using probes for the so-called sites I, II, bilirubin and fatty acids binding sites. Cephalosporins were classified into three groups according to their affinity for albumin: low affinity (K = 10 10(2) M-1), medium affinity (K = 10(3) M-1) and high affinity (K = 10(4) M-1). Cephalosporin binding to albumin produced a perturbation of several basic amino acids of the protein such as histidine and lysine. It was found that only cefuroxime, ceftazidime and cefoperazone interact slightly with site I on serum albumin, while site II possesses capacity to bind: cephradine, cephalexin, ceftazidime, ceftriaxone, cefoperazone, cefaclor and cefsulodin. The bilirubin binding site showed capacity to interact with a great number of cephalosporins: ceftriaxone, cefazolin, cephaloglycin, cefamandole, cefotaxime, cefoxitin, cefuroxime, cefoperazone and cefadroxil. Ceftriaxone showed capacity to bind to the fatty acid binding site on HSA. No relation was found between the displacement of the marker and the chemical nature of the substituents at R1 and R2. Cephalosporins interact with HSA at the binding region that involves: tyrosyl 411, histidyl 146 and lysyls 195, 199, 225, 240 and 525 residues. The chemical modification of specific amino acids showed that the interaction of these amino acids with beta lactam antibiotics is not carried out to the same extent for all the cephalosporins tested. The results obtained revealed that the binding sites for cephalosporins on albumin are structurally heterogeneous, having different amino acids in the vicinity of the ligand molecule. PMID- 9212782 TI - Palladium(II) compounds with potential antitumour properties and their platinum analogues: a comparative study of the reaction of some orotic acid derivatives with DNA in vitro. AB - Ethidium bromide was used to study perturbations induced in salmon sperm DNA complexed with a series of platinum and palladium compounds obtained from chloro and orotic acid derivatives as leaving ligands. The antitumoral activity of these compounds against Sarcoma 180 cells grafted intraperitoneally into mice is correlated with their capacity to interact with DNA in vitro and to perturb its secondary structure. Nevertheless, among these compounds, [Pt(Dach)(3-methyl orot)] and [Pt(Dach)(5-fluoro-orot)] do not interact with DNA in vitro and are inactive against Sarcoma 180 cells. This lack of activity originates from the fact that strong chelating properties of the ligand prevent hydrolysis of the compounds which are unable to give rise to aquo species which are the reactive ones. On the other hand, the interaction with DNA is not the only prerequisite in order that a compound be active towards tumour cells. In fact, cis-[Pd(NH3)2Cl2] and cis[Pd(Dach)Cl2] are not antitumoral. It is well known that the former undergoes an inactive trans-conformation and that the two compounds hydrolyse very fast assuming that they interact in vivo with a lot of molecules particularly proteins preventing them to reach the DNA, their pharmacological target. By contrast, [Pd(Dach)(3-methyl-orot)] (T/C = 267%) and [Pd(Dach)(5 fluoro-orot)] (T/C = 270%) display significant antitumour activity. PMID- 9212785 TI - Tissue-specific differences in adduct formation by hepatocarcinogenic and sarcomatogenic derivatives of 7H-dibenzo[c,g]carbazole in mouse parenchymal and nonparenchymal liver cells. AB - Parenchymal (PC) and nonparenchymal (NPC) liver cells have different tissue specific, procarcinogen activation enzymes. NPC appear to be protected against the mutagenic effects of lipotropic bulky adducts induced by carcinogens by a unknown mechanism. Most studies of activation have been conducted with whole liver. The purpose of this study was to differentiate adduct formation in mouse PC and in NPC, isolated after in vivo administration of 7H-dibenzo(c,g)carbazole (DBC), the most efficient liver carcinogen in mice, which also has potent sarcomagenic and epitheliomagenic activities. The very sensitive 32P-postlabeling method was used to detect adducts. Two tissue-specific DBC derivatives, 6-methoxy DBC (6MeODBC), which is exclusively sarcomagenic, and 5,9-dimethyl-DBC (DiMeDBC), which is exclusively hepatocarcinogenic, were analyzed in parallel. Both PC and NPC generated the ultimate metabolites of DBC, but NPC were substantially less efficient. Clear-cut tissue-specific differences in adduct formation were established: the sarcomagenic 6MeODBC gave rise only to NPC-DNA adducts, and the hepatocarcinogenic DiMeDBC only to PC-DNA adducts. The chromatograms of the adducts were compared with those of mouse embryonic cells in culture and mouse epidermal cells. The results are discussed in connection with animal experiments with DBC, 6MeODBC, and dimethylbenzanthracene and with published data on PC and NPC activating enzymes. PMID- 9212784 TI - Genotoxicity and cytotoxicity in male B6C3F1 mice following exposure to mixtures of 1,3-butadiene and styrene. AB - 1,3-Butadiene and styrene are oxidized, in part, by cytochrome P450 2E1 and have been shown to metabolically interact in rodents exposed by inhalation to mixtures of both compounds. Because the reactive metabolites of butadiene and styrene are thought to be responsible for the toxicity of each compound, metabolic interactions may alter the response in animals exposed to mixtures of butadiene and styrene compared with the response in animals exposed to butadiene alone or styrene alone. The purpose of this study was to quantitate alterations in genotoxicity and cytotoxicity in male B6C3F1 mice exposed to mixtures of butadiene and styrene. Male B6C3F1 mice were exposed to 6.25, 62.5, 200, or 625 ppm butadiene alone, 50 ppm styrene alone, or mixtures of 6.25, 62.5, 200, or 625 ppm butadiene and 50 ppm styrene. Genotoxicity was assessed by quantitating the frequency of micronucleated polychromatic erythrocytes in bone marrow. Cytotoxicity was assessed by counting total spleen and thymus cells and by quantitating the frequency of polychromatic erythrocytes in the peripheral blood. Butadiene and mixtures of butadiene and styrene were genotoxic in mice, as shown by a significant increase in the frequency of micronucleated polychromatic erythrocytes. The increased frequency following exposure to mixtures of butadiene and styrene was not significantly different compared with the frequency following exposure to butadiene alone. Styrene and mixtures of butadiene and styrene were cytotoxic in mice, as shown by significantly decreased number of spleen cells. Exposure to mixtures of butadiene and styrene with butadiene concentrations of 62.5 or 625 ppm significantly reduced the number of thymus cells. Exposure to 200 ppm or 625 ppm butadiene alone, or to mixtures of 200 ppm or 625 ppm butadiene and 50 ppm styrene, significantly reduced the frequency of polychromatic erythrocytes in the peripheral blood. The results of the study demonstrate that exposure to mixture of butadiene and styrene does not reduce the respective genotoxicity of butadiene or cytotoxicity of styrene. PMID- 9212786 TI - Dose-response relationship for the induction of chromosomal abnormalities in gamma-irradiated human spermatozoa. AB - The cytogenetic effects of in vitro irradiation on human spermatozoa have been studied by the interspecific in vitro fertilization system between human sperm and hamster oocytes. Semen samples from three healthy men were irradiated at doses of 0.00, 0.10, 0.25, 0.50, 1.00, 2.00, and 4.00 Gy. A total of 340 chromosome complements derived from non-irradiated human spermatozoa and 987 complements from irradiated spermatozoa were analyzed after sequential uniform stain-G banding. Both the frequency of spermatozoa with structural chromosome abnormalities, and the incidence of such abnormalities per cell, showed strong dose-effect relationships that were best expressed by linear-quadratic equations: Y = 0.06413(+/-0.00475) + 0.1982(+/-0.00833)D - 0.00763(+/-0.00204)D2 and Y = 0.07385(+/-0.00838) + 0.23329(+/-0.03124)D + 0.02317(+/-0.00955)D2, respectively. When analyzing separately unrejoined and rejoined structural abnormalities, we found that the incidence of unrejoined lesions was four times higher than the incidence of rejoined anomalies. The induction of unrejoined abnormalities showed a linear, dose-dependent increase, whereas the incidence of rejoined abnormalities showed a quadratic, dose-dependent increase. The distribution of radiation-induced breakpoints was also analyzed. Breakpoints were found to be randomly distributed among chromosomes, but a clustering of breakpoints in G negative bands was found: 71.5% of breakpoints were located in G-negative bands, and 28.5% in G-positive bands. PMID- 9212787 TI - Thiabendazole-induced cytogenetic abnormalities in mouse oocytes. AB - Of the various classes of human genetic disorders, aneuploidy is the most prevalent. Besides its association with maternal age and its predominant origin during maternal meiosis I, little is known about the etiology of aneuploidy. Although various classes of chemicals have been shown to induce aneuploidy in experimental systems, there is no definitive evidence for the role of chemically induced aneuploidy and adverse human health effects, particularly germ cell effects. Thus, it is important to understand the potential of chemicals for inducing aneuploidy in germ cells. There are conflicting data in the literature about the ability of thiabendazole (TBZ) to induce aneuploidy; therefore, we investigated the potential of TBZ for inducing aneuploidy in oocytes. Superovulated ICR female mice were administered 0, 50, 100, or 150 mg/kg TBZ by intraperitoneal injection. The frequencies and percentages of hyperploid oocytes were 0/472 (0), 2/410 (0.5), 6/ 478 (1.3), and 3/427 (0.7) for control, 50, 100, and 150 mg/kg TBZ, respectively. The difference between controls and the 100 mg/kg dose was statistically significant. Also, the proportions of ovulatory mice and the number of oocytes collected per ovulatory female were reduced in the TBZ groups relative to controls. Based on these results, we conclude that TBZ induces a small, but significant increase in the frequency of aneuploid oocytes at toxic doses that also impair ovulation. PMID- 9212788 TI - Comparative 32P-postlabeling analysis of exogenous and endogenous DNA adducts in mouse skin exposed to a wood-preserving waste extract, a complex mixture of polycyclic and polychlorinated chemicals. AB - Wood preserving waste (WPW) sites contain numerous toxic compounds, including phenols, polycyclic aromatic hydrocarbons (PAHs), polychlorinated dibenzodioxins, and dibenzofurans. Previous in vitro and in vivo 32P-postlabeling studies showed the induction of multiple carcinogen-DNA adducts by WPW extracts. We now have tested the hypothesis in a mouse skin bioassay that a WPW extract not only causes the formation of exogenous, xenobiotic-derived DNA adducts, but also alters the levels of endogenous DNA modifications. Skin DNA of female ICR mice treated topically with an organic WPW extract was found by 32P-postlabeling to contain significantly increased levels of bulky oxidative DNA lesions (type II I compounds), in addition to exogenous PAH-derived adducts. The mechanism of this increase is postulated to proceed through electrophilic quinoid compounds, which presumably were formed from phenols by chemical reactions of waste material or biologically by oxidative metabolism. On the other hand, the levels of another class of endogenous DNA adducts (type I I-compounds) were reduced significantly in exposed skin DNA. This effect was explained by the presence of cytochrome P450 inducers in the extract. All three types of DNA alterations observed may play a significant role in carcinogenesis. Our results imply that in addition to exogenous carcinogen-DNA adducts, alterations of endogenous DNA modifications may need to be considered in evaluating carcinogenic risk from toxic chemical wastes and the effects of remediation measures. PMID- 9212789 TI - An evaluation of micronucleus induction in bone marrow and in hepatocytes isolated from collagenase perfused liver or from formalin-fixed liver using four week-old rats treated with known clastogens. AB - The bone marrow (BM) micronucleus (MN) test is a sensitive assay for identifying clastogens. However, some clastogenic compounds and metabolites may never reach the BM. The liver has been suggested as an alternative tissue to BM but adult rat liver has a low mitotic index that increases the difficulty of evaluating hepatocytes (HEP) for MN induction. Chemical mitogens and partial hepatectomy have been used to increase HEP proliferation to improve the sensitivity for detection of clastogenic compounds, but these practices raise concerns for the evaluation of drug candidates. The use of 4-wk-old rats provides an alternative to mitogenic stimulation because livers from these animals have approximately 5.4% of their HEP in S-phase. HEP were isolated by collagenase perfusion, or from formalin-fixed tissue, from 4-wk-old treated rats. Six compounds were evaluated for the incidence of MN in HEP that were isolated by both methods. The results for MN induction by these compounds were similar for the two methods and confirmed that formalin-fixed tissue is an acceptable source of cells for evaluating MN induction in HEP. BM polychromatic erythrocytes (PCE) also were harvested at the end of the live phase for each study and then evaluated for the incidence of MN. Diethylnitrosamine and 2-nitrofluorene induced MN in HEP but had no effect in PCE. 2-Acetylaminofluorene, cyclophosphamide and 7,12 dimethylbenz[a]anthracene did not induce MN in HEP but were positive in PCE. The direct-acting clastogen, mitomycin C, was positive in both HEP and PCE. These results indicate that this modified liver micronucleus test, using 4-wk-old rats, offers an alternative to existing methods that use mitogens or partial hepatectomy to stimulate cell replication. Analysis of MN from formalin-fixed tissue provides additional flexibility by allowing the investigator to assess MN induction at a later time. PMID- 9212790 TI - Assessment of the ability of propoxur, methomyl, and aldicarb, three carbamate insecticides, to induce micronuclei in vitro in cultured Chinese hamster ovary cells and in vivo in BALB/c mice. AB - Three carbamate insecticides (propoxur, methomyl, and aldicarb) were evaluated for their ability to induce micronuclei (MN) in vitro using cultured Chinese hamster ovary (CHO) cells, and in vivo in mouse bone marrow erythrocytes. In vitro, all three insecticides induced a significant increase in micronucleated binucleate cells, which was generally both dose and sample time dependent. The in vivo studies involved treating male BALB/c mice by different routes, either once or on 3 consecutive days, followed by multiple or single sampling. Treatment by intraperitoneal injection or oral gavage induced a significant increase in micronucleated reticulocytes (MNRETs) in peripheral blood. For all three chemicals, the MN response depended on sample time and the number of treatments, while for aldicarb, the response depended also on the route of exposure. These positive results demonstrate that propoxur, methomyl, and aldicarb are capable of inducing structural and/or numerical chromosomal aberrations in mammalian cells either in vitro or in vivo. Furthermore, based on the results obtained, on optimal in vivo MN protocol for carbamate insecticides is a single treatment followed by blood sampling at 24 and 48 hr after treatment. PMID- 9212791 TI - Formation and repair of O6-methylguanine in recombination hot spots of plant chromosomes. AB - Mutagen-induced chromatid aberrations are not randomly distributed along the metaphase chromosomes. In the field bean (Vicia faba), defined late-replicating and transcriptionally inactive heterochromatic regions are preferentially involved. After exposure to the alkylating agent N-methyl-N-nitrosourea (MNU) (10(-3) M, 1 hour), 70% of all aberrations are clustered within 6 segments containing tandemly repeated FokI elements of 59 bp, which comprise approximately 10% of the genome. Using immuno-slot-blot analyses, we have studied the frequency of O6-methylguanine (O6-MeG), a mutagenic lesion important for aberration induction, in total genomic DNA as well as in FokI sequences of the field bean after exposure to MNU. In either case, similar numbers of adducts per nucleotide were found immediately after treatment as well as after 18 hours of recovery, when most adducts were removed and significant amounts of chromatid aberrations were detectable. Peculiarities of long FokI element arrays (e.g., formation of specific tertiary structures), resulting in error-prone recombination repair, rather than preferential formation or delayed repair of O6-MeG are apparently responsible for aberration clustering in these hot spot regions. PMID- 9212792 TI - Removal of O6-methylguanine from plant DNA in vivo is accelerated under conditions of clastogenic adaptation. AB - Previously it was shown that the clastogenic efficiency of high doses of alkylating agents in plant root meristems can be reduced significantly by conditioning pretreatment with either a low dose of the same agents, a sublethal heat shock, or heavy metal salts. The molecular mechanisms responsible for these protective effects are still unclear. Here we report on the quantification of O6 methylguanine [O6-MeG] by immuno-slot-blot analysis in DNA of root tip meristems of field bean (Vicia faba) seedlings under conditions of clastogenic adaptation. When root tips were pretreated with a low, conditioning dose of N-methyl-N nitrosourea (MNU, 10(-4) M, 1 hour) 2 hours before exposure to a high dose of the same clastogen (10(-3) M, 1 hour), the frequency of chromatid aberrations was reduced by more than 50% at a recovery time of 1 B hours, as compared to treatment with the high dose alone. The same was observed when conditioning pretreatment was by a sublethal heat shock [10 minutes, 40 degrees C] or a heavy metal salt (Cd(NO)3, 10(-7) M, 1 hour). The frequency of O6-MeG immediately after exposure to a conditioning and a subsequent challenge treatment was reduced by 43% as compared to treatment with only the high dose. At a recovery time of 18 hours the corresponding frequency of adducts was reduced by 68.3% (related to the initial level) after treatment with the high dose alone, and by 81.3% under adaptive conditions. Sublethal heat shock or heavy metal salt used as conditioning pretreatments also resulted in a decrease of adducts immediately after treatment with the challenge dose. From these data and from prevention of the effects by pretreatment with cycloheximide or O6-benzylguanine we conclude that under conditions of clastogenic adaptation O6-MeG is more efficiently removed from the DNA, presumably by induction of an alkyl acceptor protein such as O6-methylguanine-DNA methyltransferase [MGMT]. This could explain the observed protective effects (clastogenic adaptation. PMID- 9212793 TI - Is the white-ivory assay of Drosophila melanogaster a useful tool in genetic toxicology? AB - The white-ivory assay of Drosophila is based on the detection of reversions to wild-type phenotype of ommatidia with the white-ivory mutation. A tandem quadruplication of this gene is used in order to increase the reversion probability. Although the exact mechanism implicated in reversion is not known, revertant spots are believed to arise as a consequence of intrachromosmal recombination or related phenomena. Since the white-ivory assay has not been broadly used, the number of chemicals tested until now is still limited. In this work, we have assayed 25 chemicals belonging to several chemical groups, i.e., crosslinking agents, DNA-topoisomerase inhibitors, antimetabolites/nucleotide pool inhibitors, cyclic-adduct inducers, halogenated hydrocarbons, bulky-adduct inducers, intercalating agents, oxidative damage inducers, and a multiple damage inducer, to validate this test. Cross-linking agents, halogenated hydrocarbons, and the multiple damage inducer, dounomycin, were positive. On the contrary, the three antimetabolites/nucleotide pool inhibitors tested were negative. The other chemical groups showed disparate results, since some chemicals were positive, whereas others were negative in each group. A comparison with the results obtained in the w/ w+ and mwh/flr3 assays shows that the wi assay detects a more restricted spectrum of damages than those, although, with respect to carcinogenicity, its sensitivity (0.76, with the 62 chemicals tested until now) is similar to that estimated for the mentioned somatic assays. The conclusion of this work, then, is that the wi assay is not recommended as a general screening test, because the background reversion frequencies show a high variability among solvents, the range of lesion-recognition is lower than in the w/ w+ and mwh/flr3 SMARTs, and the mechanism implicated in the white-ivory reversion is poorly understood. PMID- 9212794 TI - Genotoxicity monitoring of small bodies of water using two species of tadpoles and the alkaline single cell gel (comet) assay. AB - To monitor genotoxicity in small bodies of water (e.g., creeks, ponds, and drainage ditches) we examined tadpole erythrocytes of two species: Rana clamitans and Rana pipiens,using the alkaline single cell gel DNA electrophoresis (SCG) or "comet" assay. This approach involves detection, under alkaline conditions, of cell DNA fragments which on electrophoresis migrate from the nuclear core, resulting in a "comet with tail" formation. Fifty-six samples, a total of 606 tadpoles, from 18 sites in southern Ontario, collected between 1993 and 1995, were examined. Samples of R. clamitans tadpoles collected in 1994 and 1995, from regions with heavy agricultural activity, gave significantly higher (P < 0.001) DNA length to width ratios than samples of R. clamitans tadpoles collected from sites in the Bruce Peninsula and near the French River, which have little or no agriculture. Samples of R. pipiens tadpoles collected in 1994 from sites on the outskirts of Windsor, Ontario, sites which receive genotoxic inputs from nearby industries, gave significantly higher (P < 0.001) DNA ratios than samples from agricultural areas and the Bruce Peninsula. R. clamitans tadpoles showed significant annual variation in DNA damage which was greater in samples of tadpoles collected from agricultural areas than from the Bruce Peninsula. The higher levels of DNA damage in tadpoles collected from agricultural areas may be due to the pesticides used, and the increased variation in DNA damage in the same areas is likely due to the impact of crop rotation, including leaving fields fallow, the timing of rainfall, and/or the application of pesticides. R. clamitans tadpoles, especially those collected from agricultural areas, also showed significant seasonal variation in DNA damage. There was no significant (P > 0.05) seasonal or annual variation in the levels of DNA damage in R. pipiens tadpoles collected from the Tallgrass Prairie. This study indicates that both species are suitable for use in the alkaline SCG assay and as in situ sentinel organisms for environmental biomonitoring. PMID- 9212795 TI - Alternate hypothesis for the bimodal size distribution of mutant colonies of L5178Y mouse lymphoma cells. PMID- 9212796 TI - Mouse lymphoma workshop: Victoria, British Columbia, Canada, March 27, 1996 protocol issues regarding the use of the Microwell Method of the Mouse Lymphoma Assay. PMID- 9212797 TI - [Cyclin and cell cycle]. AB - Cell cycle progresses through the formation of cyclin produced and degraded in its specific phase and cyclin dependent kinase (Cdk) existing in all phases of the cell cycle and consequential phosphorylation of Cdk. The Cdk is activated by Cdk-activating kinase as well as cyclin, and its function is suppressed by its inhibitory subunits (CKIs). The cell cycle induced by extracellular stimuli in normal cells progresses toward cell differentiation and apoptosis or cell proliferation according to a certain law. The failure to complete cell-cycle progression can be detected and repaired at many checkpoints. Cancer cells have abnormal cell-cycle relating constituents, such as overexpression of cyclin, mutation or deletion of CKI and failures in feedback controls at checkpoints. These abnormal constituents may contribute to the evaluation of patients' prognosis and cellular sensitivity for chemotherapy and ionizing irradiation and may lead to a development of new promising anti tumor agents. PMID- 9212798 TI - [Familial cancer and prevention project--a view on UICC symposium]. AB - Along with the marked development of molecular biology, familial cancer study has made a giant breakthrough in understanding carcino-genesis and provided an evolutional opportunity for genetic testing which has opened an encouraging scope in development of cancer prevention strategy Progress of molecular biology will be accelerated to overcome the cost effectiveness barrier in near future. However, there are no way to reduce a lag time required for the longitudinal observational study on these tested individuals not only in physical but also in psychosocial aspects, and on result of preventive intervention on their course, those which are essential indeed in achieving the aim of the research. In this regards, we urgently need to create the essential infrastructure involving construction of the permanent and nation wide cancer genetic information system, our own fundamental agreement on ELSI and decisiveness to acknowledge the need for educating and recruiting cancer genetic counselor. In this occasion, UICC symposium is the most timely event that will elicit an realistic impact in the new era of cancer research, in Japan. PMID- 9212799 TI - [Knudson's "two-hit" hypothesis in familial cancer syndromes]. AB - Cancer is a heritable disorder of somatic cells. Environment and hereditary both operate in the origin of human cancer. Hereditary cancer is rare, but it has served as a useful model in the understanding of carcinogenesis. A number of cancer genes have been identified by the study of hereditary cancers and implicated in sporadic forms of the same tumors. In this article, I reviewed Knudson's famous "two-hit" hypothesis of tumor suppression. PMID- 9212800 TI - [Genetic testing for cancer susceptibility: the present situation in Japan]. AB - Genetic diagnosis of hereditary tumors has progressed extensively and contributed to the presymptomatic testing for mutation carriers. This evolution of genetic testing for cancer susceptibility leads to improved prevention and early detection of cancers. However, the benefits and limits of testing, and the range of prevention and treatment are different in each hereditary tumor. The American Society of Clinical Oncology (ASCO) made the statement on genetic testing for cancer susceptibility and showed three categories for consideration of cancer predisposition testing. They recommend that genetic testing should be offered only for the categories including well-defined tumors. We described the characteristics of hereditary tumors in the three categories and discussed the significance of genetic testing for each tumor. In conclusion, genetic testing for cancer susceptibility should be applied only for subsets of hereditary syndromes, and it is important to continue the analysis of the significance (frequency or penetrance) of mutations of cancer predisposition genes and to make clear the genotype-phenotype and other correlations. PMID- 9212801 TI - [Current status of clinical diagnosis and DNA test of von Hippel-Lindau disease in Japan]. AB - An outline of the clinical diagnosis including the DNA testing in von Hippel Lindau (VHL) disease is briefly explained. The current status of the diagnosis and treatment of VHL disease in Japan is also compared with those in European countries. According to the current experience in the world, DNA testing of VHL disease is regarded as one of the important references for the clinical diagnosis of this disease. We have to improve various points in the Japanese status of the clinical diagnosis, genetic counseling, treatment and follow-up in patients with VHL disease. PMID- 9212802 TI - [Ethical legal and social issues in pre-symptomatic testing for cancer susceptibility in familial tumors]. AB - Recent advance in molecular biology have led to the identification several inherited cancer susceptibility genes. The pre-symptomatic testing is expected to reduce cancer morbidity and mortality by preventive intervention, early detection and adequate management. But this new predictive tests may raise ethical legal social issues (ELSI) in association with the right to control private information and confidentiality. The implications of test results are enormous, not only for the individuals but also for relatives who share this genetic legacy and society as a whole. Genetic testing for cancer susceptibility should generally be performed only within the context of long-term outcome studies which are designed to measure the medical and psychological effectiveness. The Ethical Subcommittee in Japanese Society of Familial Tumors is now elaborating the guidelines for the research on genetic testing for familial tumors in order to support the individual or family who are the subjects of the research of the clinical applications and to protect their human rights. Current standards for contents and process of informed consent and core elements in obtaining consent for DNA sample storage in medical research were listed up. PMID- 9212803 TI - [Chemoprevention of familial cancer]. AB - I review basic researches and clinical trials on chemoprevention of familial adenomatous polyposis (FAP) which is one of well-known familial cancer. Many basic researches indicate that nonsteroidal anti-inflammatory drugs (NSAIDs) prevent colorectum neoplasms by inhibition of cyclooxygenase (COX) 2. Actually sulindac, which is one of the NSAIDs, reduces adenomas of the rectum of FAP patients. I think, however, sulindac should not be used for cancer prevention, because it can not prevent advanced cancer but it reduces adenomatous polyps of the rectum. Furthermore, side effects of sulindac are frequent and serious. PMID- 9212805 TI - [Inoperable esophageal carcinoma managed by combined chemotherapy (CBDCA, 5-FU and VDS) and radiotherapy]. AB - Eleven inoperable patients with advanced esophageal carcinoma were treated with chemotherapy (carboplatin, 5-FU, vindesine) and concomitant radiotherapy. Two patients (T2) received this treatment due to their poor general condition and refusal of operation, and 9 patients for infiltration of tumor into the adjacent organs (T4). Administration of carboplatin (30 mg/body) and 5-FU (250 mg/body) together with radiotherapy (1.8 Gy/d) for 5 days a week was performed. This chemoradiation therapy was carried out for 5 consecutive weeks. In addition, vindesine (1-3 mg/body) was administered in the 1st and 4th week. After evaluation, endoscopic balloon dilatation was performed in 6 patients with stenosis of the esophagus. The general response rate was 80%. CR was noted in 2 patients of T2 but 1 patient of T4 developed severe leucopenia and immunosuppression, and died of septic MOF. All but the MOF case could take enough food orally following the endoscopic dilatation. The 1-year survival rate in the T4 group (45%) was significantly better than the non-treatment group (0%). In conclusion, this treatment is beneficial for patients with inoperable esophageal carcinoma to obtain a satisfactory QOL and survival rate. PMID- 9212804 TI - [Combination chemotherapy of continuous infusion 5-fluorouracil and daily low dose cisplatin in advanced gastrointestinal and lung adenocarcinoma]. AB - Continuous intravenous infusion (c.v.i.) of 5-fluorouracil (5-FU) plus daily low dose cisplatin (CDDP) was evaluated in 45 patients with advanced and recurrent unresected colorectal, lung, gastric and pancreatic adenocarcinoma. 5-FU was given at a dose of 320 mg/m2/day, c.v.i. for 4 weeks, and CDDP between 3.5 to 7 mg/m2/day, infused for one hour five times a week for 4 weeks. Patients received 1 to 3 cycles of treatment (average 1.5 cycle). Pancreatic cancer cases needed longer treatment periods (2.25 cycles). The response rate of colorectal cancer cases was 57.7% (15/26), pancreas cancer 40%, gastric cancer 62.5%, and lung cancer 66.7%. The overall response rate was 57.8%. No severe side effects occurred in any of these cases. These data indicate that this combination 5-FU + daily low-dose CDDP chemotherapy is effective in the treatment of advanced gastrointestinal and lung adenocarcinoma. PMID- 9212807 TI - [Study of disposition of adriamycin and mitomycin C in liver by determination of plasma concentrations in hepatic vein and artery during intravenous constant infusion in rats]. AB - The hepatic extraction ratios (EH) of adriamycin (ADR) and mitomycin C (MMC), which were administered clinically by an intra-hepatic arterial route, were measured in rats to clarify the disposition of ADR and MMC in liver. EH values of ADR and MMC were determined by comparing the femoral arterial and hepatic venous plasma concentrations at steady state during continuous intravenous administration. The EH value of ADR in rats at each infusion rate of 2, 10 and 50 micrograms/kg/min, was 0.290, 0.286 and 0.251, respectively. There was no significant difference between the EH values (p > 0.05). The systemic clearance (CLtot) at each infusion rate was 108, 77.6 and 72.9 ml/min/kg, respectively. The EH value of MMC in rats at each infusion rate of 2.5, 7.5 and 25 micrograms/kg/min, was 0.332, 0.358 and 0.360, respectively. There was no significant difference between the EH values, the same as for ADR. The systemic clearance (CLtot) at each infusion rate was 38.3, 36.1 and 35.3 ml/min/kg. PMID- 9212806 TI - [Pyrimidine nucleoside phosphorylase activity, 5-fluorouracil concentration and thymidylate synthase inhibition rate in colorectal cancer after oral administration of 5'-doxifluridine]. AB - To assess the mechanism of the anticancer effect of doxifluridine (5'-DFUR), a clinicopharmacological study was performed. So far, no comparative study has been reported between the thymidylate synthase (TS) inhibition rate and the 5-FU concentration in colorectal cancer after oral administration of 5'-DFUR. In 37 patients with colorectal cancer, 5'-DFUR was administered orally/preoperatively (800 mg/day x more than 4 days before operation and 300 mg on the day of operation). The mean total dosage was 6.9 g. Specimens of tumor and normal intestinal tissue were obtained 4.4 hours on average after final administration. The TS inhibition rate as well as the 5-FU concentration and the activity of pyrimidine nucleoside phosphorylase (PyNPase) were analyzed in both tissues. The PyNPase activity was significantly higher in the tumor tissue than in the normal tissue (137.9 +/- 10.8 vs. 31.0 +/- 4.7 micrograms FU/mg protein/hr, p < 0.0001). The 5-FU concentration was also significantly higher in the tumor tissue than in the normal tissue (101.3 +/- 30.6 vs. 23.2 +/- 5.5 ng/g, p = 0.024). The TS inhibition rate correlated with the 5-FU concentration in the tumor tissue (r = 0.527, p = 0.047). These findings suggest that the TS inhibition rate may be an index of the anticancer effect of 5'-DFUR in colorectal cancer. PMID- 9212808 TI - [The usefulness of fibrin glue spraying combined with CDDP for lung cancer]. AB - The local slow release of an anticancer agent by one route is a form of treatment. There have been numerous reports to date that this approach might be a promising adjuvant for systemic therapy. We investigated the spraying of the operative field at a local site with cisplatin, which is considered the most effective carcinostatic in the field of lung cancer, and reported it in clinical cases selected in applying a Pilot-Study. The feature of this method is to use fibrin to spray drugs uniformly over a wide field. We expect it will be an adjuvant for postoperative purification and other uses. PMID- 9212809 TI - [Clinical evaluation of 2 mg granisetron tablet for nausea and vomiting induced by anticancer drugs including cisplatin]. AB - 1. The clinical efficacy and safety of the 2 mg granisetron tablet were assessed in 32 mainly lung cancer patients who were to receive treatment with anticancer drugs including CDDP. 2. One 2 mg granisetron tablet was administered prophylactically one hour before the start of CDDP administration. 3. Based on the development of nausea and vomiting in 24 hours after the start of CDDP administration, the study medication was judged to be "remarkably effective" or "effective" in 71.0% (22/31) of cases. 4. The study medication was judged to be "safe" in 96.9% (31/32) of cases, without causing any adverse reactions. 5. The above results indicate that the 2 mg granisetron tablet is safe and useful. PMID- 9212810 TI - [Clinical efficacy of GG032X tablets, a new dosage form of ondansetron (fast dispersing tablet), on cisplatin-induced nausea and emesis]. AB - The inhibitory effects of GG032X tablets, a new dosage form (fast dispersing tablet) of ondansetron, 5-HT2 receptor antagonist, on nausea and emesis induced by cisplatin (CDDP), were investigated along with safety and usefulness. Subjects were chemotherapy patients starting CDDP administration for the first time, who were receiving a high single dose of CDDP (50 mg/m2 or more and intravenous drip infusion of less than 4 hours), or lower multiple doses of CDDP (a single dose of 10 mg/m2 or more, administered intravenously for 3-5 consecutive days). GG032X tablets were administered orally 1-2 hours before CDDP administration. In lower multiple doses of CDDP, GG032X tablets and CDDP were administered, as much as possible, at the same respective time when they were administered on the first day. Efficacy of GG032X tablets was evaluated in terms of inhibitory effect on nausea and emesis 24 hours after administration of a high single dose of CDDP, and of the inhibitory effect on nausea and emesis during the study period (3-5 days) in lower multiple doses of CDDP. Efficacy, safety and usefulness were evaluated in accordance with the evaluation criteria used in the clinical study of already-approved ondanstron tablets. In a high single dose of CDDP, the cases judged "effective" or better in the investigation of the inhibitory effect of the drug on nausea and emesis, accounted for 52.9% (63/119 cases). As for the overall safety rating, the cases judged as "safe" accounted for 87.0% (107/123 cases), and as a "minor safety problem" accounted for 13.0% (16/123 cases). As for the usefulness rating, the cases judged "useful" or better accounted for 52.1% (62/119 cases). Major adverse effects included headache, fever, atrial fibrillation and increases in total bilirubin, GOT and GPT values. None of these was serious, and the patients recovered without any treatment or by nosotropic therapy. Meanwhile, in lower multiple doses of CDDP, the inhibitory effect judged "effective" or better accounted for 70.6% (12/17 cases). As for the overall safety rating, all cases were judged "safe". In terms of usefulness, those cases judged "useful" or better accounted for 70.6% (12/17 cases). No adverse effect was observed. Study results of these two groups were almost the same as those for already-approved ondansetron tablets. According to the results of questionnaires for the patients who participated in the study and took GG032X tablets, the drug was found to be easy to take and had favorable results. Based on the above results, GG032X tablets were evaluated as having the same inhibitory effect as the already-approved ondansetron tablets against CDDP-induced nausea and emesis, and were considered safe and clinically useful. PMID- 9212811 TI - [Two cases of adenosquamous cell carcinoma of advanced cervical cancer treated by carboplatin-based chemotherapy with peripheral blood stem cell autotransplant]. AB - In two cases with adenosquamous cell carcinoma of advanced cervical cancer, carboplatin-based chemotherapy was given intraarterially from the internal iliac artery as neoadjuvant chemotherapy, and peripheral blood stem cells (PBSCs) were harvested. After the operation, conventional intravenous chemotherapy with PBSC autotransplant was performed. PBSCs were mobilized by neoadjuvant chemotherapy and G-CSF administration. By the apheresis procedures, 0.7-2.6 x 10(6)/kg CD34 positive cells were obtained. They had no severe side effects from intravenous chemotherapy with PBSCT, and they were free of disease 20 months. Neoadjuvant chemotherapy and G-CSF administration may be capable of mobilization of PBSCs, and chemotherapy with PBSCT may be useful in radioresistant advanced adenosquamous carcinoma of the cervical cancer. PMID- 9212812 TI - [A case of postbulbar duodenal ulcer due to adjuvant chemotherapy after gastrectomy for gastric cancer]. AB - We encountered a rare case of postbulbar ulcer caused by adjuvant chemotherapy after gastrectomy. A 64 year-old woman, who received chemotherapy with CDDP-5-FU after gastrectomy, developed severe hematemesis. Endoscopic examination revealed a gigantic ulcer at the anal site of Vater's papilla. The ulcer was healed with PPI and soft diet. PMID- 9212813 TI - [An anaphylactic shock case after hepatic arterial infusion of zinostatin stimalamer suspension improved by anti-histaminics]. AB - A 47-year-old man with hepatocellular carcinoma (HCC) at anterior and medical segment in the liver was treated with hepatic arterial infusion of Zinostatin Stimalamer-lipiodol suspension (SMANCS). After the 2nd infusion of SMANCS, the accumulation of lipiodol in the tumor was not good (Grade II), so additional administration was undertaken at five-weeks intervals. His systolic blood pressure immediately decreased from 120 to 60 mmHg, and he had numbness of hands, shaking chills, sweating, chest pain and numerous urticaria-like red exanthema. In spite of treatment by anti-shock agents such as steroid and catecholamines, these symptoms did not disappear, but antihistaminics greatly improved them without any serious side effects. Because of the remarkable effects of the antihistaminics and possibility of antibody production (IgE) after repeated infusions of high molecular SMANCS, this patient may have suffered anaphylactic shock caused by massive histamine release from mast cells. PMID- 9212814 TI - [A case of unresectable pancreatic cancer successfully treated with continuous venous daily infusion of 5-FU and low-dose CDDP]. AB - This case was a 61-year-old male diagnosed with pancreatic cancer who had a celiotomy. The tumor invasion involved the pancreas body and head. Ultrasonograph examination revealed that all the celiac artery, supramesenteric artery, and portal vein were occluded in the tumor. Because of the unresectability of the tumor, a palliative operation was carried out, and during the following 9 months he was given UFT 300 mg daily. Because abdominal ascites accompanied the tumor growth, the patient underwent combination chemotherapy of cisplatin 5 mg/day x 5/week and continuous infusion of 5-fluorouracil 250 mg/day for 4 weeks. Remarkable reduction of the abdominal ascites and decline of the tumor markers (CEA, CA19-9) was observed in the course of the chemotherapy. Bone marrow function was suppressed by the agents, but granulocyte colony stimulating factor (G-CSF) was very effective for recovery from the damage. Two months after discharge, abdominal ascites recurred. The patient received the same serial chemotherapy for 6 weeks, and now is followed as an outpatient. PMID- 9212816 TI - [TNM classification of bone and soft tissue sarcomas]. AB - TNM classification of bone and soft tissue sarcomas was published by UICC in 1987. Histological grading (G) is an important factor in this classification, but the criteria of G categories are not so clear. In addition, lymph node metastasis is very rare in bone and soft tissue sarcoma. Therefore, prognostic factors are limited to T, M and G categories. Since correlation between the stage (UICC) and the survival rate was not found in patients with osteosarcoma, TNM classification (UICC) has not been used widely in the field of orthopedic oncology. The Musculoskeletal Tumor Committee of the Japanese Orthopaedic Association proposed another TNM classification of osteosarcoma based on multivariate analysis. T1 is less than 15 cm and T2 is 15 cm or larger in maximal diameter. N and M are same with the UICC criteria. Serum alkaline phosphatase level (A) is included in this classification in which A0 is less than the normal value x2.5, and A1 is the normal value x2.5 or more. G categories are separated into two groups according to the mitotic rate in a high power field (x200); G1 is assigned to the tumor with 0-9/1 HPF and G2 is assigned to those with 10 or more/1 HPF. Reclassification of osteosar-coma by this modified TNM system indicated that there was a correlation between the survival rate and the stage. PMID- 9212815 TI - [Secondary hepatic resections in a case of sigmoid colon cancer with multiple liver metastasis (H3) after successful continuous hepatic artery infusion chemotherapy oriented by in vitro chemosensitivity test]. AB - A 43-year-old woman was admitted to our hospital for sigmoid colon cancer with multiple liver metastasis (H3). As preoperative CTAP (CT during arterial portography) examination showed 23 metastatic nodules in the whole liver, hepatic resections were not indicated. Angiographic findings showed that right and left hepatic arteries branched separately from the celiac artery. Sigmoid colon resection with D3 lymph node dissection and catheterization to the right hepatic artery via gastroduodenal artery were undertaken as a first operation. Continuous hepatic artery infusion chemotherapy with MMC, 5-FU oriented by in vitro chemosensitivity test (SDI test: Succinic Dehydrogenase Inhibition test) of primary tumor was performed 7 days after the first operation. After administration of MMC (40 mg) and 5-FU (16,500 mg), metastatic nodules in the right lobe almost disappeared except for the one tumor of S7, but the size and number of the nodules in the left lobes increased. At 10 months after the first operation, the left hepatic lobectomy and extirpation of only one tumor in the right lobe (S7) underwent. This case showed the usefulness of continuous hepatic artery infusion chemotherapy oriented by in vitro chemosensitivity test for multiple liver metastasis from colon cancer. PMID- 9212817 TI - Immunochemical characterization of a human sperm fibrous sheath protein, its developmental expression pattern, and morphogenetic relationships with actin. AB - Among the monoclonal antibodies (MAbs) prepared against human sperm extracts, MAb 4F7 was found to be specific to the human and Macaca fascicularis sperm cytoskeletal fibrous sheath (FS). In Western blotting, MAb 4F7 stains a doublet of polypeptides of about M(r) 95 x 10(3) in extracts of human sperm cells. These polypeptides are not recognized by the KL1 anti-cytokeratin MAb, nor by the MAbs known to bind to the carboxy terminal (IFA) and to the amino terminal (ME101) rod domain of intermediate filaments. Sequential extraction procedures shows that the FS polypeptides recognized by MAb 4F7 are exposed after treatment with 8 M urea 4F7 immunoreactivity is lost after treatment with high ionic solutions (NaCl; KCl, Kl). Immunogold electron microscopy reveals that this protein is present throughout the FS. This FS antigenic determinant first accumulates in an FS proximal body in late spermatids, then in granules extending distally along the flagellum. Staining of spermatozoa with flagellar dysgenesis reveals that this FS protein colocalizes with actin no matter what the location of their abnormal assembly. These data suggest that the transient microtubule-like spindle-shaped body of as yet unknown function could be involved in FS protein deposition and that the assembly of the FS and actin could be under the control of some common morphogenetical factor(s). MAb 4F7 should allow further investigations of this peri-axonemal structure in both normal and pathological conditions. PMID- 9212818 TI - Major DNA fragmentation is a late event in apoptosis. AB - Apoptosis, the terminal morphological and biochemical events of programmed cell death, is characterized by specific changes in cell surface and nuclear morphology. In addition, DNA fragmentation in an internucleosomal pattern is detectable in mass cultures of apoptotic cells. However, DNA fragmentation and nuclear morphological changes may not necessarily be associated events. In this study, we examined OVCAR-3 and KB human carcinoma cells using time-lapse video phase-contrast microscopy to characterize the surface and nuclear morphological features of apoptosis in response to treatment with either taxol or ricin. The surface morphological features of apoptosis were the same in both cell types and with both drugs. Using an in situ nick-translation histochemical assay, these single cells were also examined for DNA strand breaks during apoptosis. Surface morphological changes demonstrated discrete stages of cell rounding, surface blebbing, followed by cessation of movement and the extension of thin surface microspikes, followed much later by surface blistering and cell lysis. Nuclear features examined by DAPI cytochemistry demonstrated apoptotic nuclear condensation very early in this sequence, usually at the time of initial surface blebbing. The nick-translation assay, however, demonstrated DNA strand breaks at a much later time, only after the formation of separated apoptotic bodies or after final cell lysis. This study points out the differences between surface and nuclear morphological changes in apoptosis, and the large temporal separation between nuclear morphological changes and major DNA fragmentation detectable by this in situ technique. This result suggests caution in using in situ nick translation as a direct correlate of internucleosomal DNA fragmentation in apoptosis. PMID- 9212819 TI - Detection of intracellular interleukin-8 in human mast cells: flow cytometry as a guide for immunoelectron microscopy. AB - The chemokine interleukin-8 (IL-8) mediates infiltration and adhesion of neutrophils during inflammatory processes. We have previously shown that this cytokine can be produced and released by normal and leukemic human mast cells (HMC-1 cells). To assess whether and to what extent this cytokine is stored intracellularly, we investigated production and localization of IL-8 at the single-cell level by combined use of flow cytometry (FACS) and immunoelectron microscopy. Conditions necessary for optimal fixation and permeabilization of HMC 1 cells were determined by measuring changes in cell-specific light scatter parameters and by estimating cellular uptake of propidiumiodide (PI). In this way, we were able to detect IL-8 with a monoclonal antibody in stimulated cells that were microwave-fixed with a combination of paraformaldehyde (4%) and glutaraldehyde (0.1%), followed by permeabilization with saponin (0.025%). FACS analysis revealed time-dependent synthesis of IL-8 with at most 50% positively stained cells at 8-12 hr after stimulation. For pre-embedding immunogold electron microscopy, cells were treated according to the protocol established by flow cytometry. IL-8 was found to be located in specific cytoplasmic, electron-dense granules of stimulated HMC-1 cells. These results confirm and extend our previous findings by demonstrating IL-8 expression in HMC-1 cells at the single-cell level. In addition, we propose that quantitative FACS can be reliably used in a timesaving manner to establish appropriate conditions for pre-embedding immunoelectron microscopy of intracellular antigens. PMID- 9212820 TI - A covalent fluorescent-gold immunoprobe: simultaneous detection of a pre-mRNA splicing factor by light and electron microscopy. AB - Immunoprobes that combine a fluorescent label with a 1.4-nm gold cluster compound have been prepared by covalent conjugation with polyclonal antibody Fab' fragments. These new immunoconjugates allow the collection of two complementary sets of data, from fluorescence and electron microscopy, from a single labeling experiment. We find that incorporation of one or more fluorescein moieties into the coordinated ligands of the 1.4-nm Nanogold gold cluster label yields a stable, dual-function immunolabel in which fluorescence quenching is negligible. In a second synthetic strategy, Nanogold and fluorescein were separately covalently conjugated to Fab' fragments to yield a probe with very similar properties. With the use of Fab' fragments, the entire probe is smaller than a whole IgG molecule, and it exhibited excellent penetration properties. It was used to localize the pre-mRNA splicing factor SC35 in the HeLa cell nucleus by both fluorescence and electron microscopy. PMID- 9212821 TI - Differential expression of CD66a (BGP), a cell adhesion molecule of the carcinoembryonic antigen family, in benign, premalignant, and malignant lesions of the human mammary gland. AB - CD66a, also known as biliary glycoprotein (BGP), is a member of the carcinoembryonic antigen (CEA) family and the human homologue of the rat cell CAM. There is evidence that aberrant expression or loss of CD66a in tumor tissue is of biological significance. No data about its expression in breast carcinoma cells and only sparse information about the expression of CD66a in normal breast are available thus far. In this study we used monoclonal antibodies to analyze the expression of CD66a and CEA in normal tissue, benign lesions, and in noninvasive and invasive carcinomas of the mammary gland. In normal tissue and benign lesions, CD66a was consistently expressed at the apical sites of epithelial cells and in myoepithelia, whereas CEA was absent or was restricted only to some apical membranes within the ductal tree. The specific staining of myoepithelia was most evident in pseudoinfiltrative radial scars and sclerosing adenosis. However, the apical expression of CD66a disappeared with the development of the malignant phenotype in noninvasive and invasive carcinomas, and changed gradually from low- to high-grade noninvasive carcinomas into a predominant uniform membrane staining all around the atypical cells. CEA expression was irregular in intensity and distribution. The native apical CD66a staining was partially preserved in some highly differentiated invasive carcinomas with a better prognosis, such as tubular and papillary carcinomas. These findings indicate that loss of CD66a expression rather than a change in staining patterns coincides with the development of the malignant phenotype. PMID- 9212822 TI - Immunolocalization of rap1 in the rat parotid gland: detection on secretory granule membranes. AB - The objective of this study was to localize rap1 in the rat parotid gland. Rap1 is a small GTP-binding protein that has been linked to phagocytosis in neutrophils and various functions in platelets. In this study, we used [alpha 32P]-GTP-blot overlay analysis, immunoblot analysis, and immunohistochemistry to identify rap1 in rat parotid gland. The immunohistochemical techniques included immunoperoxidase and widefield microscopy with image deconvolution. Rap1 was identified in the secretory granule membrane (SGM), plasma membrane (PM), and cytosolic (CY) fractions, with the largest signal being in the SGM fraction. The tightly bound vs loosely adherent nature of SGM-associated rap1 was determined using sodium carbonate, and its orientation on whole granules was assessed by trypsin digestion. Rap1 was found to be a tightly bound protein rather than a loosely adherent contaminant protein of the SGM. Its orientation on the cytosolic face of the secretory granule (SG) is of significance in postulating a function for rap1 because exocytosis involves the fusion of the cytoplasmic face of the SG with the cytoplasmic face of the PM, with subsequent release of granule contents (CO). Therefore, the localization and high concentration of rap1 on the SGM and its cytosolic orientation suggest that it may play a role in the regulation of secretion. PMID- 9212823 TI - Tomography of cells by confocal laser scanning microscopy and computer-assisted three-dimensional image reconstruction: localization of cathepsin B in tumor cells penetrating collagen gels in vitro. AB - We used the nondestructive procedures of confocal laser scanning microscopy in combination with computer-assisted methods to visualize tumor cells in the process of penetrating collagen gels. Three independent sets of images were collected. The image information of all data sets was combined into one image, giving a three-dimensional (3D) impression at high light microscopic resolution and sensitivity. We collected information about the extracellular matrix using the reflection mode, the cell surface/morphology by staining with the fluorescent dye DiOC6(3), and the distribution of cathepsin B by Cy-3-labeled immunolocalization. The specific aim of our study was visualization of the spatial relationship of cell organelles as far as they contain the enzyme cathepsin B to cell morphology and motility in a 3D model of extracellular matrix. The majority of the enzyme was localized pericellularly, with no visible relationship to the direction of movement. However, substantial amounts also appeared in intramatrix pseudopodia and associated with the extracellular face of the plasma membrane, which may be indicative either of secretion and/or epicellular activity. Our approach has general applicability to study of the spatial relationships of cell compartments and their possible reorganization over time. This could open new horizons in understanding cell structure and function. PMID- 9212825 TI - Co-localization of endogenous and exogenous p53 proteins in nasopharyngeal carcinoma cells. AB - Recently, we have established nine nasopharyngeal carcinoma (NPC) cell lines in which only one cell line showed the p53 mutation. For investigation of the p53 mutation in this line, immunostaining using anti-p53 antibody was applied and showed the presence of p53 protein in the cytoplasm but not in the nucleus. Single strand conformation polymorphism analysis of the p53 gene showed one normal and one additional DNA band. Cloning and sequencing of PCR-amplified DNA showed an AGA (arginine) to ACA (threonine) heterozygous point mutation at codon 280. Transfection of the p53 DNA binding sequence and chloramphenicol acetyltransferase assay revealed loss of transcriptional activation function of endogenous p53 protein. Co-localization of the endogenous and the transfected exogenous p53 protein by polyclonal antibodies to anti-p53 protein revealed strong exogenous p53 staining in the transfected nuclei and weak staining of endogenous p53 protein in the cytoplasm. We concluded that (a) a heterozygous point mutation at codon 280 was identified in the NPC-TW 06 cell line; (b) the point mutation may cause the stagnation of mutant p53 protein in the cytoplasm, and loss of its transcriptional activation function; (c) endogenous and exogenous p53 protein can be co-localized at the same time in the transfected cells; and (d) 280 mutant p53 protein in NPC cells does not cause a decrease or increase in sensitivity to chemotherapy. PMID- 9212824 TI - Production and characterization of specific anti-peptide antiserum against free alpha-subunit of rat pituitary glycoprotein hormones. AB - To obtain an antibody specific for the alpha-subunit of rat pituitary glycoprotein hormones, we synthesized a peptide corresponding to the sequence 37 53 (ST-7: Phe-Ser-Arg-Ala-Tyr-Pro-Thr-Pro-Ala-Arg-Ser-Lys-Lys-Thr-Met-Leu-Val) of the rat alpha-subunit. The polyclonal antiserum against this peptide was generated in rabbits. This region is hydrophilic and highly conserved among several mammalian species. Noncompetitive binding tests showed that the ST-7 antiserum had specific affinity for the rat free alpha-subunit, but not for rat intact LH, FSH, and TSH. The ST-7 antiserum immunostained two types of cells in the rat anterior pituitary, i.e., gonadotrophs and thyrotrophs. This was also the case in mouse, cattle, sheep, and pig, which have an identical sequence of ST-7 in their alpha-subunit. The pituitary cells of horse (Arg substituted for Lys as residue 48 of the rat alpha-subunit), human, and eel (Leu for Ala at residue 45), chicken (Met for Ala at residue 45), and bullfrog (Tyr for Phe at residue 37 and Met for Ala at residue 45) were not stained with the ST-7 antiserum. This study indicated that the ST-7 antiserum is sequence-specific for the alpha-subunit and is therefore useful for immunohistochemical studies on the secretory pathway of the free alpha-subunit. PMID- 9212826 TI - Differential expression of the fibroblast growth factor receptor (FGFR) multigene family in normal human adult tissues. AB - This report describes a systematic analysis of the expression of the fibroblast growth factor receptor (FGFR) multigene family (FGFR1, FGFR2, FGFR3, and FGFR4) in archival serial sections of normal human adult tissues representing the major organ systems, using immunohistochemical techniques. Polyclonal antisera specific for FGFR1, FGFR2, FGFR3, and FGFR4 and a three-stage immunoperoxidase technique were employed to determine the cellular distribution of these receptors at the protein level. The expression profiles for the tissue-specific cellular localization of the FGFR multigene family demonstrated wide-spread and striking differential patterns of expression of individual receptors in the epithelia and mesenchyme of multiple tissues (stomach, salivary glands, pancreas, thymus, ureter, and cornea) and co-expression of FGFR1-4 in the same cell types of other tissues. The wide-spread expression of FGFR1-4 in multiple organ systems suggests an important functional role in normal tissue homeostasis. Differences in the spatial patterns of FGFR gene expression may generate functional diversity in response to FGF-1 and FGF-2, both of which bind with equally high affinity to more than one receptor subtype. In vivo, this may lead to functional differences that are crucial for the regulation of normal physiological processes and are responsible for the pathological mechanisms that orchestrate various disease processes. PMID- 9212827 TI - Spatiotemporal expression of C-CAM in the rat placenta. AB - We investigated the expression of the immunoglobulin superfamily cell adhesion molecule, C-CAM, in developing and mature rat placenta. By immunohistochemical staining at the light microscopic level, no C-CAM-expression was seen before Day 9 of gestation, when it appeared in the trophoblasts of ectoplacental cones. On Day 10.5, spongiotrophoblasts and invasive trophoblasts around the maternal vessels of the decidua basalis were stained positively. On Day 12.5, C-CAM was detected in the spongiotrophoblasts of the junctional layer, but labyrinth trophoblasts and secondary giant trophoblasts were not stained. On Day 17.5, C CAM was found only in the labyrinth and lacunae of the junctional layer. At this stage, both the labyrinth cytotrophoblasts of the maternal blood vessels and the endothelial cells of the embryonic capillaries were strongly stained. Placental tissues from gestational Days 12.5 and 17.5 were analyzed by immunoelectron microscopy to determine the location of C-CAM at the subcellular level. On Day 12.5, positive staining of the spongiotrophoblasts was observed, mainly on surface membranes and microvilli between loosely associated cells. On Day 17.5, staining was found primarily on the microvilli of the maternal luminal surfaces of the labyrinth cytotrophoblasts, and both on the luminal surface and in the cytoplasm of endothelial cells of the embryonic vessels. RT-PCR analysis and Southern blotting of the PCR products revealed expression of mRNA species for both of the major isoforms, C-CAM1 and C-CAM2. Immunoblotting analysis of C-CAM isolated from 12.5-day and 14.5-day placentae showed that it appeared as a broad band with an apparent molecular mass of 110-170 kD. In summary, C-CAM was strongly expressed in a specific spatiotemporal pattern in trophoblasts actively involved in formation of the placental tissue, suggesting an important role in placental development. In the mature placenta, C-CAM expression was confined to the trophoblastic and endothelial cells lining the maternal and embryonic vessels, respectively, suggesting important functions in placental physiology. PMID- 9212828 TI - N-ethylmaleimide (NEM) can significantly improve in situ hybridization results using 35S-labeled oligodeoxynucleotide or complementary RNA probes. AB - We predicted that a significant source of background labeling after in situ hybridization (ISH) using 35S-labeled probes is attributable to a chemical reaction between the phosphorothioate moiety of the probe [O3P = S] and disulfides in tissue. These covalent bonds would immobilize probe in the tissue, thereby increasing background labeling. On the basis of this view, we have explored the use of N-ethylmaleimide (NEM) to irreversibly alkylate the phosphorothioate moiety of the probe and/or to alkylate free sulfhydryls in tissue to block the formation of disulfides as a method of reducing background labeling. We report that NEM can significantly decrease background labeling of 35S-labeled oligodeoxynucleotide or cRNA probes but does not affect specific labeling. We conclude that the use of NEM in ISH protocols, as outlined here, may be an additional element researchers may consider to improve the signal-to-noise ratio. PMID- 9212829 TI - Liposomes and immunoassays. AB - Various aspects of the application of liposomes as a label in immunoassays are reviewed. Methods for the preparation of liposomes, from the basic film method to the more advanced dehydration-rehydration method, are discussed. Furthermore, the markers used in liposome labels, as well as the methods to conjugate liposomes to antigens or antibodies, are summarized. Liposome immunoassays are applied as homogeneous or heterogeneous assays. Homogeneous assays often rely on the lytic activity of complement on antibody-associated liposomes. Another group of homogeneous assays utilizes the inhibitory action of antibodies on the activity of conjugates of mellitin (a bee venom protein) with a hapten. Free mellitin conjugates are able to lyse liposomes effectively. Heterogeneous liposome immunoassays, performed either competitively or non-competitively, resemble more closely standard enzyme linked immunosorbent assays, with the enzyme being replaced by a liposome label. Washing steps are used to separate antigen specifically bound liposomes from unbound liposomes. All bound liposomes are lysed with a detergent, giving an instantaneous amplification. Flow-injection liposome immunoassays and liposome immunosensors are also described as examples of other possible immunoassay formats. PMID- 9212830 TI - Analyzing cytotoxic T lymphocyte activity: a simple and reliable flow cytometry based assay. AB - A flow cytometry-based assay for analyzing cytotoxic T lymphocyte (CTL) activity is presented. This new approach is characterized by easy handling, the generation of highly reproducible data sets and is not dependent on the use of radioactivity. Before exposure to primed CTL effector cells the target cells were labeled with the green fluorescent dye DiO18(3) which is incorporated stably into the cell membrane. After a 4-h incubation period, samples were counterstained with the red fluorescent nuclear dye propidium iodide in order to permit discrimination between live and dead cells within both cell populations. The assay has been used to quantitate CTL effector activity against allogeneic lymphoblasts. Results derived from this novel flow cytometry assay show an excellent correlation (r = 0.988) with data obtained using the standard 51chromium release assay. An additional advantage of the assay is that freshly prepared splenocytes may be used as target cells because culturing and activation of target cells is no longer required. The results demonstrate that the fluorescent dyes DiO18(3) and propidium iodide in combination with flow cytometry permit accurate analysis of cytotoxic T cell activity. PMID- 9212831 TI - Extremely efficient gene transfection into lympho-hematopoietic cell lines by Epstein-Barr virus-based vectors. AB - We have estimated the efficiency of Epstein-Barr virus (EBV)-based vectors in transfecting genes into cell lines of lympho-hematopoietic lineages. The transfection efficiency was estimated both at transient and stable phases, in terms of expression of a marker gene and acquisition of drug resistance, respectively. Plasmid vectors carrying EBV oriP (replication origin of plasmid), EBNA (EBV nuclear antigen)-1 and as the marker genes, murine CD8 alpha cDNA and neoR (neomycin resistant) genes were transfected into various cell lines by electroporation. When cell lines constitutively expressing EBNA-1 were transduced, virtually all the cells expressed CD8 alpha on day 3 and acquired G418 resistance thereafter. In the case of K562 cells, which do not express EBNA 1, approximately 40% of cells expressed the marker gene product on day 3 posttransfection, and 30% of cells became stable transfectants. These data suggest a broader application of the EBV vector system in basic immunology and medicine. PMID- 9212832 TI - Modification of surface marker expression on CD14 monocytes of allergic patients after lysis or Ficoll purification. AB - It has been observed that peripheral blood monocytes are often in a primed or activated state in inflammatory diseases such as asthma. However, the majority of these studies have been performed using cells which have been purified by density gradient centrifugation on Percoll or Ficoll-Hypaque. Using cytofluorimetry, we compared the expression of monocyte surface markers of monocytes from untreated blood with monocytes purified by erythrocyte lysis or density centrifugation using the Ficoll technique. Monocytes from two groups of subjects were analyzed: healthy subjects and allergic patients. When compared with untreated blood, the percentage of CD16-positive cells, and the sMFI was significantly greater after monocyte purification (lysis or Ficoll). The expression of CD62L (percentage and sMFI) was modified after monocyte purification. Such modification of these two surface markers was predominantly observed on monocytes from allergic patients, and not on monocytes from healthy subjects. This study suggests that surface marker analysis should be performed on unfractionated whole blood in order to avoid modification of monocyte antigens. PMID- 9212833 TI - Inorganic pyrophosphatase-labelled enzyme immunoassay for beta 2-microglobulin. AB - beta 2-Microglobulin was purified from human peritoneal dialysate by ultrafiltration, gel chromatography and DEAE-high performance chromatography. Anti-beta 2-monoclonal antibodies were developed in mice and a pair of the antibodies was utilized to develop an enzyme-linked immunosorbent assay (ELISA) kit for the analyte with utilizing a new enzyme label, inorganic pyrophosphatase (EC 3.6.1.1). The sensitivity of the assay was 0.6 microgram/l (3 x SD) and the assay was linear up to absorbance values of around 2.0. No hook effect occurred in any putative concentrations of beta 2-microglobulin in serum. The precision of the assay of one run varied within 5-7% CV and the interassay precision was 2.8 8.6% CV, while the recovery was 99.2 +/- 6.0%. Excellent correlation occurred against an established radioimmunoassay method. All the used reagents were storable for a minimum of 1 year at +4 degrees C. It was decided that inorganic pyrophosphatase is a label of choice for ELISA. PMID- 9212835 TI - How should CD34+ cells be analysed? A study of three classes of antibody and five leucocyte preparation procedures. AB - For patients undergoing stem cell transplantation after intensive marrow ablative therapy it is important to enumerate the CD34+ stem cells in peripheral blood so that the harvest can be timed in order to maximize the number of cells collected by leucophoresis for subsequent haematopoietic reconstitution. The use of rapid flow cytometric techniques for the determination CD34+ leucocyte numbers has been advocated, although there is no consensus as to the best method. In this study, we have examined the effects of preparation procedures for flow cytometry on the binding of four CD34 antibodies (Immu-133, QBEND-10, HPCA2 and BIRMA-K3) to the three classes of epitopes on leucocytes. Whole blood, bone marrow and leucophoresis samples were analysed either directly after labelling with a vital nuclear dye (LDS-751) and fluorochrome-conjugated antibodies or after additional erythrocyte lysis and leucocyte fixation using four commercially available reagents (Q-Prep, OptiLyse B, OptiLyse C and FACS Lysing Solution). By comparison with the results obtained from viable leucocytes in unmanipulated samples, it was found that the binding of all four antibodies could be affected by lysis and fixation procedures and that the binding of the class I antibody Immu-133 was most markedly decreased. We conclude that CD34+ cells are best analysed using a whole blood procedure in which nucleated cells are identified by their side light scatter and the fluorescence associated with a vital nuclear dye (in this instance LDS-751) and the CD34+ cells are detected with fluorescein isothiocyanate- or phycoerythrin-conjugated antibodies. PMID- 9212834 TI - A monoclonal antibody reacts with maltose-binding protein of Escherichia coli and related enteric bacteria. AB - Maltose-binding protein (MBP) encoded by malE is essential for the energy dependent translocation of maltose through the cytoplasmic membrane of bacteria. Its property of specific binding to maltose has been used in constructing fusion proteins for easy affinity purification. A monoclonal antibody named MAb SC1D7 was produced against Escherichia coli MBP. This MAb also bound to MBP-containing recombinant proteins in both Western blotting and immunoprecipitation analysis. As a result, this MAb can be a useful probe for tracing MBP-fusion proteins in various applications. Furthermore, intrinsic MBPs from E. coli, Shigella dysenteriae, Salmonella typhimurium, Enterobacter cloacae, and Klebsiella pneumoniae were also detected by this MAb. No reaction was observed with the total proteins from Serratia marcescens, Aeromonas hydrophila and Plesiomonas shigelloides. These observations suggest that the MAb SC1D7-defined epitope is conserved among some enteric bacteria, but not the others. The results strengthen the phylogenetic positions of these closely related bacteria previously placed by other means. PMID- 9212836 TI - Human endothelial culture supernatant selectively promotes IgM-producing hybridomas. AB - We compared the effect of human endothelial cell culture supernatant (HECS), ESG (Ewing sarcoma growth factor), IL-6 (interleukin-6) and peritoneal macrophages on the recovery of hybridoma cells after fusion with respect to growth, stability and distribution of isotype variants. A selective growth of murine IgM-producing hybridoma cells was observed in the presence of HECS after cell fusion. PMID- 9212837 TI - Methodology for selection of human antibodies to membrane proteins from a phage display library. AB - We describe a simple antigen capture technique for the selection of a specific human antibody to p185erbB-2, a transmembrane glycoprotein, from a library of human Fab genes expressed on the surface of bacteriophage. Magnetic beads coated with the rat antibody ICR55 have been used to capture erbB-2 antigen from Triton X-100 extracts of SKOV3 cells. The antigen-coated beads have then been used to select bacteriophage displaying human Fab with affinity for p185erbB-2. After 4 rounds of selection, 65 phage clones were isolated which bound specifically to p185erbB-2 in a capture assay. Nine of the clones which gave the strongest reaction in an ELISA were selected for further development and the Fab genes were subcloned into the expression vector pUC119his6mycXba and electroporated into E. coli TG1. Colonies were grown, induced and the supernatants tested for the presence of secreted human Fab. Supernatants from two of the 9 clones contained human Fab and one of these bound specifically to erbB-2 in a capture assay, stained the membranes of the erbB-2 overexpressing cell lines BT474 and SKBR3 and immunoprecipitated a protein of molecular weight 185 000 kDa from SKOV3 cells. We conclude that a membrane antigen captured by specific monoclonal antibody can be used successfully to select phage displaying human antibodies specific for the antigen. PMID- 9212838 TI - Assessment of the Alamar Blue assay for cellular growth and viability in vitro. PMID- 9212839 TI - Contemporary classification of histiocytic disorders. The WHO Committee On Histiocytic/Reticulum Cell Proliferations. Reclassification Working Group of the Histiocyte Society. AB - Pathologists and pediatric hematologist/ oncologists of the World Health Organization's Committee on Histiocytic/Reticulum Cell Proliferations and the Reclassification Working Group of the Histiocyte Society present a classification of the histiocytic disorders that primarily affect children. Nosology, based on the lineage of lesional cells and biological behavior, is related to the ontogeny of histiocytes (macrophages and dendritic cells of the immune system). Dendritic cell-related disorders of varied biological behavior are dominated by Langerhans cell histiocytosis, but separate secondary proliferations of dendritic cells must be differentiated. Juvenile xanthogranuloma represents a disorder of dermal dendrocytes, another dendritic cell of skin. The hemophagocytic syndromes are the most common of the macrophage-related disorders of varied biological behavior. Guidelines for distinguishing the exceedingly rare malignant diseases of histiocytes from large cell lymphomas through the use of a battery of special studies are provided. PMID- 9212840 TI - Systemic hemophagocytosis masking the diagnosis of large cell non-Hodgkin lymphoma. AB - An 11-year-old female presented with clinical features suggestive of malignant histiocytosis: fever, weight loss, subcutaneous nodules, pulmonary infiltrates, adenopathy, and hepato-splenomegaly. On biopsy, lymph node and bone marrow demonstrated necrosis and extensive hemophagocytosis with no definitive evidence of malignancy: the subcutaneous nodules, however, demonstrated large-cell non Hodgkin lymphoma. This clinicopathologic picture has been reported in adults, but not in children. Although serum G-CSF, M-CSF, and TNF levels were not elevated in this child, it is possible that other cytokines induced either directly or indirectly by the subcutaneous lymphoma resulted in hemophagocytosis. PMID- 9212841 TI - Undifferentiated sarcomas of children: pathology and clinical behavior--an Intergroup Rhabdomyosarcoma study. AB - Undifferentiated soft tissue sarcoma (UND-STS) is the most poorly defined tumor eligible for intergroup Rhabdomyosarcoma Studies (IRS). Recent IRS UND-STS experience was reviewed to assess the histologic characteristics and clinical behavior of undifferentiated sarcomas. Of the 1,527 patients entered on IRS-III and IRS pilot-IV, 96 had tumors classified by the IRS Pathology Committee as UND STS. Of these, 52 had adequate histologic material for this study. After application of immunohistochemistry, 18 tumors were reclassified, mostly as embryonal rhabdomyosarcomas (RMS), primitive neuroectodermal tumors, and intraabdominal desmoplastic small found cell tumors. The remaining 34 UND-STS had a diffuse hypercellular histologic pattern made up of sheets of medium-sized cells. The tumor cells had a minimal to moderate amount of cytoplasm and a variable nuclear morphology, predominately vesicular with finely granular chromatin. Except for reactivity with antibodies against vimentin, most tumors had a negative immunohistochemical profile. The 5 year Kaplan-Meier survival estimate for patients with non-metastatic disease was 72%, a significant improvement when contrasted with patients diagnosed to have UND-STS in IRS-I and IRS-II. PMID- 9212842 TI - Primary and metastatic rhabdomyosarcoma in the breast: neoplasms of adolescent females, a report from the Intergroup Rhabdomyosarcoma Study. AB - The occurrence of rhabdomyosarcoma (RMS) primary in or metastatic to breast has been regarded as an uncommon event, associated with an unfavorable outcome. Records of 26 patients with diagnoses of breast RMS, either primary or secondary, entered in the Intergroup Rhabdomyosarcoma Study (IRS) (1972-1992) were reviewed and compared with data regarding 47 similar patients in published reports. Of the 26 IRS cases, the histologic subtype was alveolar in 24, embryonal in 1, and not determined in 1. All were female with ages ranging from 11.5 to 20.2 years (median, 15.2 years; mode, 14-16 years). This compact age distribution of both primary (n = 7) and metastatic (n = 19) breast RMS was seen in previously reported series. Among the 19 cases of RMS with initial dissemination to breast, primary tumor sites, were extremity (n = 8), nasopharynx/paranasal sinuses (n = 7), and trunk (n = 4). IRS treatment was risk-based according to site and extent of disease. Four of 7 patients with primary RMS remain disease free 2.9 to 7 years post diagnosis. Among 19 patients with RMS initially metastatic to breast, including 7 in IRS clinical group IV at original diagnosis, three are disease free at 7.6, 15.7 and 17.0 years. CONCLUSIONS: primary or metastatic RMS in breast is almost confined to adolescent females having tumors with alveolar histology. Approximately one-half of the patients with primary breast disease and 15% of those with metastatic breast disease as an initial recurrence are long term survivors. PMID- 9212844 TI - Trends of survival in neuroblastoma and independent risk factors for survival at a single institution. AB - To assess the progress of survival in neuroblastoma which varies with many risk factors and to evaluate the influence of these factors on survival as independent risk factors. The study subjects were 159 neuroblastoma patients seen from 1965 1994 at the oldest and largest children's hospital in Japan. Trends of survival in three treatment eras-1965-81, 1982-86, 1987-94-were assessed by the Kaplan Meier method for different sex, age at diagnosis, the clinical stage, the site of onset, and the histological type. Then the influence on survival of these factors as independent prognostic variables was evaluated by the Cox proportional hazards regression analysis. Age at diagnosis, the clinical stage, the site of onset, the histological type, and the treatment era were independent risk factors in the order of their influence on survival. Unfavorable survival outcomes were obtained for patients with age at diagnosis above 1 year, the clinical stage of VI by the Evans classification, adrenal onset, and neuroblastoma rather than ganglioneuroblastoma. Survival improved from the first to the second and from the second to the third treatment era. Improvement of survival in neuroblastoma took place during the past 3 decades. Age at diagnosis, the clinical stage, and the histological type have still remained overwhelming prognostic factors over the progress in treatment. PMID- 9212843 TI - Multimodal therapy for children and adolescents with Ewing sarcoma: results of the First National Chilean Trial (1986-1991). AB - Thirty-seven patients with Ewing sarcoma were treated in the First National Chilean Trial for Ewing's Sarcoma (1986-1991), which comprised the St. Jude Ewing's 78 Study. All patients received cyclophosphamide, doxorubicin, vincristine, and Dactinomycin for a total treatment period of about 10 months, and all prescribed therapy was administered. Local therapy consisted of irradiation (RT) to the primary tumor, complete surgical resection, or a combination of both surgery and RT. Twenty-nine of these patients had localized tumors, 24% had pelvic primary tumors, 21 were males, and 20 were greater than 10 years of age at diagnosis. Twenty-one patients had tumors that were greater than 8 cm in largest diameter. Fourteen of the 29 patients with localized disease remain disease free at 23 to 91 months from diagnosis. Fourteen patients have died of-tumor-related complications and 1 of a secondary malignancy. Relapse was local only in 4, metastatic in 9, and local plus metastatic in 1. Only 1 of the 8 patients with metastatic disease at presentation remains disease free. Toxicity consisted primarily of myelosuppression and mucositis. We conclude that this form of relative intense multimodal therapy for children/adolescents with localized Ewing sarcoma is curative in about half of affected children as in the original St. Jude study, and that it can be safely given in a developing country, provided that careful attention to supportive care and treatment planning is given. Although these results represent improvement in outcome for our patients, more effective therapy is needed for children with Ewing sarcoma, especially those with metastatic disease at presentation. PMID- 9212845 TI - Concomitant p53 mutation and MYCN amplification in neuroblastoma. AB - The MYCN oncogene is amplified in 20% of childhood neuroblastoma and is associated independently with poor prognosis. Alteration of the p53 tumor supressor gene, in contrast, occurs infrequently in these tumors. In this report, we described a 3-year-old girl with stage IV neuroblastoma. Molecular analysis revealed, both MYCN gene amplification and a point mutation of the p53 tumor supressor gene. To our knowledge, this is the first reported case of neuroblastoma with genetic alterations of both these genes. PMID- 9212846 TI - Incidence of malignant neoplasms in children attending Social Security Hospitals in Mexico City. AB - An increase in neoplasms in Mexican children has been reported. In 1991, the incidence in children from Mexico City (MC) was 70 (x 10(6) child/year), although this rate might be underestimated. The aim of the present study was to estimate the incidence of malignant neoplasms in children resident in MC attending Social Security (SS) hospitals. This study was a retrospective hospital survey. All records of childhood malignant neoplasms diagnosed between 1992 and 1993 in the two SS hospitals which attend childhood neoplasms in MC were reviewed. Histopathological diagnoses were reevaluated and incidence rates (x 10(6) child/ year) in terms of age, sex, and place of residence were estimated. A total of 667 cases were found for the period of study, of which 199 corresponded to residents of MC. The neoplasms with highest prevalence were leukemias (39.2%), lymphomas (17.6%), and central nervous system tumors (12.6%). A general incidence of 94.3 was found, which was highest in children under 5 years of age. Leukemias had an incidence of 36.4, lymphomas of 15.2, and central nervous system tumors of 12.0. Prevalence was higher in boys (male/female ratio of 1.6). As for the place of residence, the highest incidence corresponded to children living in the southern areas of MC. Eighty percent of the leukemias were acute lymphoblastic, while 54% of solid neoplasms were classified as stages III and IV. In conclusion, the incidence of malignant neoplasms in children resident in MC treated at SS hospitals is consistent with that found worldwide, and also with the Latin American pattern. PMID- 9212847 TI - Antioxidant micronutrients and childhood malignancy during oncological treatment. AB - Serum antioxidant vitamins A (retinol) and E (alpha-tocopherol), beta-carotene, zinc, and selenium, and cholesterol and related proteins for 170 children with newly diagnosed malignancy were measured at diagnosis and 6 months after initiation of treatment, and compared with those of 632 cancer-free controls. Incident cancer cases and controls were 1-16 years old and recruited between 1986 and 1989. At diagnosis, age- and sex-adjusted serum concentrations of retinol, beta-carotene, zinc, and alpha-tocopherol were significantly inversely associated with cancer. No significant decreases in mean values were observed at 6 month, except for the alpha-tocopherol-to-cholesterol ratio in patients with bone tumors and serum zinc in bone tumors and central nervous system malignancies. An increase during the period of treatment was found for retinol and selenium in leukemia patients. beta-carotene was maintained at the initial concentrations determined prior to therapy. These findings provide further information about micronutrient requirements in children with cancer. PMID- 9212848 TI - Tissue polypeptide-specific antigen in pediatric patients: assessment of normal values. AB - Measurement of serum concentrations of tissue polypeptide-specific Antigen (TPS) has been demonstrated to the useful in diagnosis and monitoring of adult epithelial tumors. So far, no data have been available on normal or pathologic TPS values in children. Therefore, the present study was designed to evaluate the normal values of TPS in childhood. Using a commercial enzyme linked immunosorbent assay (ELISA) kit, serum TPS was determined in 361 healthy children. Median (M) TPS was found to be 107 U/l at birth (n = 124). By the end of the first week, the value rose to M = 150 U/l (n = 68) and then continuously decreased with age (1 week-1 year, n = 45, M = 88 U/l; 1-7 years, n = 75, M = 51 U/l) until reaching the adult level (8-18 years, n = 49, M = 34 U/l). Additionally, the serum TPS values of 45 mothers right after delivery (M = 161 U/l) were assessed, and there was no correlation to the marker levels determined in the cord blood of their children. The age-dependent distribution of serum TPS in healthy children must be taken into account in the clinical application of this tumor marker. PMID- 9212850 TI - Portal vein thrombosis in association with factor V Leiden mutation in a patient with hepatocellular carcinoma. AB - We report a case of hepatocellular carcinoma associated with portal vein thrombosis. Analysis of whole cellular DNA demonstrated heterozygosity for the factor V Leiden mutation. The patient also had marked protein C deficiency. The presence of the mutation associated with protein C deficiency may increase the risk of thrombosis in this patient with hepatocellular carcinoma. PMID- 9212849 TI - Postoperative limited volume irradiation in a child with a solitary brain metastasis from Wilms tumor: a case report. AB - An 11-year-old boy presented with a solitary cerebral metastasis 2.5 years after initial diagnosis and 4 months after successful combined modality treatment of a stage II recurrent Wilms tumor in the chest. Resection of the brain metastasis was followed by limited volume irradiation with 30.0 Gy total dose. After a follow-up of 2.5 years the boy is in complete remission and shows no neurological or neuropsychological deficits indicating the possibility of curative postoperative radiotherapy of low toxicity and restricted to the tumor site in the presence of solitary brain metastases. PMID- 9212851 TI - Cutaneous toxicity following the administration of dactinomycin. AB - Dactinomycin (AMD) is an effective drug in the management of several malignant disorders and has been used for almost 40 years. Skin and subcutaneous toxicities following extravasation are well known and can be harmful. Similarly radiation recall is a well established phenomenon following the administration of AMD. We report a patient who developed a localized brawny erythema in the crural folds and the axillae, likely due to AMD. This rare skin complication of AMD seems to benefit from topical corticosteroid treatment, although postinflammatory hyperpigmentation may take months to disappear. PMID- 9212852 TI - Mediastinal endodermal sinus tumor. PMID- 9212854 TI - "Response to Hord and Janco re chemotherapy for unresectable pancreatoblastoma". PMID- 9212855 TI - Development of a test battery (NPM-X) for neuropsychological and neuromotor examination of children with developmental disabilities or mental retardation. A theoretical and clinical study. AB - Biological and behavioural diagnosis often do not provide information on functional competence. This is, however, of utmost importance in planning services as well as in research on treatment effects for children with developmental disorders. For school-aged children neuropsychological assessment has proved its value in this respect. For children of chronological age (CA) below 5-7 with specific developmental disabilities, and for children with severe mental retardation there has been a lack of applicable test batteries. This thesis presents a new test battery for neuropsychological and neuromotor examination, NPM-X, for these two groups of children. The first part of the thesis reviews available medical and psychological tests and assessment procedures with respect to applicability and relevance for neuropsychological assessment to children with mental retardation and mental age (MA) below 7. The second part describes the theoretical background and the content of the new test battery. The methodology for testing these children, who due to their age and/or their developmental disabilities often co-operate poorly, is described. Scoring categories, specifically developed to enable a detailed and differentiated description of the child, are presented. Because of the instability of the behavioural function in early age as well as in cases of severe disability, the scoring system records both the child's optimal functional capacity and inconsistencies in behaviour. For the purpose of planning treatment and training according to the child's resources as well as dysfunctions, two different functional profiles are provided. In the normative functional profile the child's functional level is compared to normal expectations for the child's CA, and in the ideographic functional profile the child's function in each area is compared to the child's average functional level. In the third part of the thesis the reliability results are presented and discussed. A pair of trained M.D.s, or psychology or special education Ph.D.s examined 110 children in a blind design. The study showed satisfactory interrater and test-retest reliability. In the fourth part current validation theory is reviewed before content and construct validity for the test battery is discussed. A concurrent criterion validation study is presented as well. Assessments available in the psychological and psychiatric records (PPR) of 35 children with CA below 7 were compared to test results obtained with NPM-X. The comparison showed high agreement in areas of function assessed both by PPR and NPM-X. In addition, NPM-X provided more information about the child's functional capacity, of relevance for the diagnostic appraisal as well as for the treatment of the child. It is concluded that a reliable and valid test battery for neuropsychological and neuromotor developmental assessment has showed its applicability and clinical utility for children with specific developmental disabilities and CA < 7 and for children with general developmental disabilities up to CA 12-13 but with MA < 7. PMID- 9212857 TI - Pathophysiology of perennial allergic rhinitis. AB - Allergic rhinitis involves an early phase, largely mediated through mast cells, and a late phase which involves cellular infiltration and mediator release. In the early phase, mast cells release mediators as a result of antigen cross linking adjacent immunoglobulin E molecules bound to mast cell surfaces. This results in an accumulation of histamine which gives rise to the characteristic symptoms of rhinitis--sneezing, itching, rhinorrhoea and congestion. The late phase of the allergic response (hours after challenge) involves infiltration of the nasal epithelium by eosinophils, basophils, monocytes and T-lymphocytes, which release leukotrienes, kinins, histamine and a host of other mediators. The most important part of the late-phase response is probably mediated via the production of cytokines (IL-4, IL-5, IL-6, IL-8, GM-CSF and RANTES) by mast cells, TH2 lymphocytes or epithelial cells. The infiltration of tissues by cells normally present only in the blood is brought about by the production of adhesion molecules, such as VCAM-1 and E-selectin, which cause circulating eosinophils, basophils and T-lymphocytes to adhere to endothelial cells before moving through the endothelium into the tissue (diapedesis). Neuronal reflexes also play a role in the allergic response, both by mediating local responses to mediators and possibly playing a part in the activation of T-lymphocytes. The allergic response has also been shown to be less intense in a hot, humid environment, and more marked in a cold, dry environment, possibly due to changes in osmolality of the nasal surface fluid. Similar factors may play a role in the aetiology of non allergic rhinitis. PMID- 9212853 TI - "Bad-risk childhood pleuropulmonary blastoma: does chemoradiotherapy help?". PMID- 9212858 TI - Autonomic reflexes and non-allergic rhinitis. AB - Functions traditionally ascribed to the nose include warming, humidification, and filtration of inspired air prior to its passage to the lower respiratory tract. The nose thus conditions inspired air, making it suitable for pulmonary gas exchange. In order to carry out these functions, the nose is subject to a complex series of reflexes mediated via the autonomic nervous system. The effector tissues are the nasal blood vessels and glands. Perennial non-allergic rhinitis may be divided into two types based on the presence or absence of eosinophilia in nasal secretions. Much circumstantial evidence suggests that non-eosinophilic non allergic rhinitis (NENAR) may be a disease of autonomic imbalance. In a series of studies carried out at the University of Liverpool, patients with perennial non allergic rhinitis, and NENAR in particular, were found to have qualitative and quantitative abnormalities in their nasal response to various stimuli as manifested by changes in nasal patency. The nasal response to axillary pressure is much reduced in NENAR patients compared with normal controls, and the normal decrease in nasal resistance in response to standing is abrogated. Isometric exercise has little effect in normal subjects, but those with NENAR demonstrate an increase in nasal resistance. A similar effect is seen in response to the cold pressor test. Of great importance for therapy is the effect of topical fluticasone propionate in patients with NENAR. Treatment normalizes the damaged nasal reflexes seen in this condition, whereas placebo has no effect. However, abnormalities in other non-nasal autonomic reflexes (systemic parameters) are not affected by treatment. PMID- 9212859 TI - The link between the nose and lung, perennial rhinitis and asthma--is it the same disease? AB - Perennial rhinitis and asthma are clinical syndromes representing a range of overlapping pathologies; accurate classification should therefore precede any comparison. Although the sinonasal cavities, trachea and bronchi have a common respiratory mucosa, there are also anatomical differences. For example, the nose has a capacitance vessel network and the lower airways possess smooth muscle, both of which are responsive to neurohumoral influences. The prevalence of rhinitis and asthma has increased over the last three decades. Rhinitis occurs in around 75% of allergic asthmatics while 20% of perennial allergic rhinitics develop asthma. Eosinophils, and their associated proteins and cytokines, may play a central role in both perennial rhinitis and asthma with and without atopy. The characteristic pathology of asthma can be summarized as a chronic, desquamating, eosinophilic bronchitis. Non-allergic rhinitis with eosinophilia is recognized, but without consistent evidence of epithelial damage. Eosinophils are also present in rhinosinusitis with polyposis, particularly in patients with aspirin sensitivity, in whom asthma also often occurs. Increased mast cell activation and mediator release is evident in both perennial rhinitis and asthma following allergen challenge. The importance of mast cells in non-atopic asthma and polyposis is also recognized. Adhesion molecules may also be upregulated, with an increased number and activation of TH2 lymphocytes. However, allergen resultant T-cell activation may be less marked in the nose than in the lung. Autonomic imbalance also plays a role in both conditions via changes in neural tone to effector tissues, release of neuropeptides, and interplay with cellular recruitment. Pharmacological manipulation of rhinitis and asthma also illustrates the pathological similarities and differences. PMID- 9212860 TI - Allergic rhinitis and inflammation: the effect of nasal corticosteroid therapy. AB - Topical corticosteroids have proved to be effective in the treatment of allergic rhinitis. The symptomatology of allergic rhinitis is considered to be the result of the accumulation and activation of inflammatory cells and cytokine release and hence the efficacy of corticosteroids is associated with their anti-inflammatory action. New advances in allergic inflammation now suggest that not only mast cells and eosinophils but also T-lymphocytes and antigen-presenting dendritic cells, play an important role in the inflammatory reaction. The effect of topical fluticasone propionate on cellular infiltration in the nasal mucosa is examined, with an emphasis on two studies performed in Rotterdam, The Netherlands. The cells influenced most by corticosteroid therapy were Langerhans' cells (antigen presenting cells), which were almost completely eradicated, possibly resulting in diminished antigen presentation, and eosinophils. There was a reduction in the number of epithelial mast cells, but the number of T-lymphocytes only decreased following high doses of corticosteroid therapy or long-term treatment. However, T lymphocyte function was influenced, as shown by the reduction in the T-helper2 (TH2)-related cytokines, interleukin (IL)-4 and IL-5. Topical corticosteroid therapy had no effect on the accumulation of macrophages. The reduction in antigen presentation, and the decrease in T-lymphocyte stimulation and cytokine production, may cause a reduced influx of eosinophils and other inflammatory cells, resulting in diminished symptomatology. PMID- 9212861 TI - The pharmacological basis for the treatment of perennial allergic rhinitis and non-allergic rhinitis with topical corticosteroids. AB - The currently available respiratory topical corticosteroids are all effective at reducing the nasal symptoms of itch, sneezing, rhinorrhoea and obstruction associated with allergic rhinitis. The mechanism of action of corticosteroids is related to their anti-inflammatory activities. They have been documented to prevent fluid exudation and reduce the number of circulating inflammatory cells, including lymphocytes, mast cells, basophils, eosinophils, macrophages, and neutrophils. This occurs through multiple mechanisms, e.g. eosinophil infiltration is suppressed by preventing cytokine production, reducing local mechanisms of tissue infiltration, and decreasing eosinophil survival. Furthermore, corticosteroids also reduce preformed and newly-generated mediators (e.g. histamine, tryptase, prostanoids, leukotrienes), and inhibit production of cytokines and chemokines by inflammatory cells (e.g. IL-1 through IL-6, IL-8, RANTES, TNF-alpha, IFN-gamma and GM-CSF). The currently available corticosteroids differ pharmacologically. Fluticasone propionate appears to have the greatest affinity for the glucocorticoid receptor, and binds more quickly and dissociates more slowly from the receptor compared with other corticosteroids, suggesting a more prolonged duration of action. Its increased specificity for respiratory tissue may lead to greater potency with less potential for systemic adverse effects. Fluticasone propionate has been compared with other corticosteroids in animal models for relative topical and systemic potency, and according to these data, it has the most favourable risk-benefit ratio. PMID- 9212862 TI - Flexible monitoring: mobilizing critical care. PMID- 9212863 TI - Engineered cell lines as a tool for monitoring biological activity of hormone analogs. PMID- 9212864 TI - Linkup of a fast protein liquid chromatography system with a stirred thermostated cell for sterile preparation of liposomes by the proliposome-liposome method: application to encapsulation of antibiotics, synthetic peptide immunomodulators, and a photosensitizer. AB - The proliposome-liposome method is based on the conversion of the initial proliposome preparation into a liposome dispersion by dilution with the aqueous phase. This technique is characterized by an extremely high entrapment efficiency and is suitable for the encapsulation of a wide range of drugs with different water and alcohol solubility. A description of a home-made stirred thermostated cell and its linkup with an FPLC system for a rapid and automated preparation of multilamellar liposomes under strictly controlled conditions (temperature, dilution rate, and schedule) is presented. The highly reproducible procedure yields multilamellar liposomes with a high encapsulation efficiency for various drugs. Carboxyfluorescein, as a model hydrophilic compound, was entrapped with an efficiency of 81 +/- 2%. The antibiotics neomycin and gentamycin were entrapped with efficiencies of 65 and 69%, respectively. Synthetic immunomodulators adamantylamide dipeptide, muramyl dipeptide, and beta-D-GlcNAc-norMurNAc-L-Abu-D isoGln were entrapped with efficiencies of 87, 62, and 85%, respectively. The photosensitizer mesotetra-(parasulfophenyl)-porphin was entrapped with an efficiency of 65%. The cell has been designed for laboratory-scale preparation of liposomes (300-1000 mg of phospholipid per run) in a procedure taking less than 90 min. The method can be readily scaled up and linked with secondary processing methods, such as pressure extrusion through polycarbonate filters. PMID- 9212865 TI - Time-resolved fluorescence microscopy could correct for probe binding while estimating intracellular pH. AB - Estimation of intracellular pH by fluorescence ratiometry overcomes many of the limitations such as variations in the pathlength of observation and concentration of the probe, light scattering, and photobleaching. However, binding of probes to membranes and macromolecules is generally not taken into account. By using time resolved fluorescence microscopy on a variety of cell types, we have shown that the dual-emission fluorescent pH probe carboxy SNARF-1 binds to cellular components in significant levels. The bound population could be resolved in the timescale since its fluorescence lifetime (approximately 3 ns) is significantly larger than that of the free probe. Intracellular pH was estimated from the relative amplitudes corresponding to free probes. This procedure was validated in simple model systems where carboxy SNARF-1 was present in solutions of bovine serum albumin. It was shown that the intracellular pH could be overestimated by as much as 1 pH unit in the absence of correction for probe binding. PMID- 9212866 TI - Engineering of functional chimeric protein G-Vargula luciferase. AB - Luciferase of Vargula hilgendorfli is infinitely stable at room temperature in dried state, and its light-emitting reaction is very simple. These unique characteristics of Vargula luciferase have prompted us to engineer chimeric protein, the other moiety chosen for conjugation being streptococcal protein G. A single domain of protein G which binds to IgG of a wide range of species was fused at the N-terminal region of Vargula luciferase. Unexpectedly, we found that the chimeric protein expressed in mammalian COS-1 cells had no IgG-binding ability, probably due to some sort of interaction between the two moieties or some conformational preferences of the IgG-binding domain of protein G when fused to Vargula luciferase. Here we report how we regained the IgG binding of protein G, by the intervention of three alpha-helices of protein A between protein G and luciferase. To our knowledge, the new chimeric protein provides the first reported model of this kind. PMID- 9212867 TI - Theoretical analysis of protein concentration determination using biosensor technology under conditions of partial mass transport limitation. AB - The theory of a method for determining active concentration of nonpurified protein samples using the BIAcore biosensor technology has been developed. The method relies on change in binding rate with varying flow rate at high ligand concentration where mass transport from bulk to surface with immobilized ligand becomes partially rate limiting. Prior study of binding kinetics is not necessary. If the molecular weight and the diffusion coefficient of the analyte protein are known, no standard with a known concentration is required. From numeric computer simulations a simple analytical expression for the mass transport coefficient derived at totally mass transport limiting conditions is shown to be applicable for conditions of partial mass transport limitation. Simple one to one association is assumed in deriving the method, but it is applicable even with more complex kinetics due to a bivalent analyte or heterogeneity of analyte or ligand. PMID- 9212868 TI - Concentration measurement of unpurified proteins using biosensor technology under conditions of partial mass transport limitation. AB - Using biosensor technology, it is possible to measure protein concentration when the binding of the protein to an appropriate ligand immobilized on the sensor surface is totally limited by diffusion and mass transport, a condition difficult to achieve in practice. In such a case, the observed binding rate does not reflect the intrinsic binding capacity of the molecular partners, but is simply proportional to the concentration of the protein analyte that is introduced in a continuous flow over the ligand. We describe here a more general biosensor method for measuring protein concentration which is applicable to conditions where mass transport is not totally but only partially rate limiting. The proposed method, which is based on measurements at different flow rates, does not require a standard of known protein concentration and can be used with unpurified proteins. The method is applicable to ligand-analyte pairs with an association rate constant as low as 10(3) M-1 s-1 and requires only knowledge of the molecular weight and diffusion coefficient of the analyte. The method was used successfully to measure the concentration of monoclonal antibodies, monoclonal antibody fragments (Fab) obtained by papain cleavage, and recombinant Fab fragments of widely different affinities in crude Escherichia coli extracts. PMID- 9212869 TI - Three-way curve resolution applied to the study of solvent effect on the thermodynamic and conformational transitions related to the protonation of polycytidylic acid. AB - Solvent effect on the acid-base behavior of polycytidylic acid [poly(C)] is studied by means of potentiometric and spectrometric (UV and CD) procedures. Low polarity biological environments are mimicked by using a 30% (v/v) dioxane-water mixture. Experiments performed in this medium are compared with previously reported results in aqueous solution in order to determine changes in both thermodynamic and structural aspects associated with modifications of the biomolecular surroundings. Potentiometric and spectrometric studies reveal the presence of a nonlinear polyelectrolytic effect associated with the protonation of poly(C) in the hydroorganic mixture, different from the analogous effect in aqueous solution. The curve resolution method alternating least squares is applied to the poly(C) spectrometric data. Concentration profiles and spectra of both deprotonated poly(C) and half-protonated [poly(C).poly(CH+)] are thus obtained. The fully protonated species poly(CH+) precipitates in the hydroorganic medium at pH values lower than 4. Evidence for the ordered structure of both poly(C) and [poly(C).poly(CH+)] species is seen through the comparison of the macromolecule spectra with those of the cytidine-3',5'-cyclic monophosphate monomer. Polarity decreases around the macromolecule produce significant hypochromicity in the CD spectra and hyperchromicity in the UV spectra, both signs of a disordering effect in the macromolecular structure due to the weakening of base stacking interactions. PMID- 9212870 TI - The quantification of protein amino groups by the trinitrobenzenesulfonic acid method: a reexamination. AB - Trinitrobenzenesulfonic acid (TNBSA) is the reagent in a well-known method for quantification of primary amino groups, but even at room temperature, N trinitrophenylation of primary amines is concomitant with hydrolysis of the reagent. The production of picric acid, the product of TNBSA hydrolysis, lowers the sensitivity of the method. Heating accelerates hydrolysis more than condensation on amino groups. The optimal pH range is centered on the value 10. The optical density is generally evaluated at 340 or 420 nm. The former value corresponds to the maximal absorption of the final product, N trinitrophenylamines. The latter value is relative to the Mesenheimer pi-complex, the well-known intermediate of the overall reaction. Both wavelengths are suitable for quantification of amines, but 420 nm seems to be the best. Further addition of sulfite is not necessary. The quantification of amines is somewhat hampered by compounds such as area and sodium dodecyl anions. The relative rates of reaction of diaminoacyl groups of protein with TNBSA differ depending on the substitution degree of the neighboring groups. Some of them do not react. The trinitrophenylation kinetic seems to be more dependent on protein structure than reactivity. In evaluation of the available lysine in glycated proteins, the distinction between reductive alkylation and Maillard-type condensation necessitates quantitative evaluation of nonalkylated lysine in protein hydrolysate, compared to results with the TNBSA method. Our results are confirmed by complete guanidization of the protein and subsequent determination of homoarginine in hydrolysates. PMID- 9212871 TI - Determination of acetyl-CoA enrichment in rat heart and skeletal muscle by 1H nuclear magnetic resonance analysis of glutamate in tissue extracts. AB - The contribution of a 13C-enriched substrate to the acetyl-CoA pool in animal tissues is typically measured by analysis of glutamate enrichment from tissue extracts. 13C NMR analysis offers the advantages of minimal sample processing and high information content, but has a low analytical sensitivity compared to other methods of tracer analysis such as GC/MS. We present a sensitive, simple, and direct 1H NMR measurement of glutamate C4 enrichment from tissue extracts. The method is demonstrated with heart and hindlimb muscle tissue extracts of rats infused with [2,4,6,8-13C4]-octanoate, a source of [2-13C]acetyl-CoA. Glutamate C4 enrichment in extracts of individual hindlimb soleus muscles weighing approximately 150 mg and containing approximately 0.3 mumol of glutamate was quantified by 1H NMR within about 40 min. Glutamate C4 enrichment measurements by 1H NMR in heart and gastrocnemius muscle were also highly correlated with independent measurements obtained from 13C NMR isotopomer analysis. PMID- 9212872 TI - A method for preparation of amino acid thiohydantoins from free amino acids activated by acetyl chloride for development of protein C-terminal sequencing. AB - A novel and efficient method to prepare amino acid thiohydantoins, which are required as reference standards for development of C-terminal protein sequencing, is reported. Amino acid thiohydantoins were prepared using a straightforward method involving reaction of 20 free amino acids with acetyl chloride as activating reagent and trimethylsilyl isothiocyanate (TMS-ITC) as derivatizing reagent. The products were characterized by HPLC, uv spectra, amino acid analysis, MS, and NMR. Different reaction conditions were investigated and the chemical mechanism of the formation of amino acid thiohydantoins was illustrated. PMID- 9212873 TI - Determination of rate and equilibrium constants for the reactions between electron transfer mediators and proteins by linear sweep voltammetry. AB - Redox proteins undergo measurable charge transfer at electrodes only under special circumstances, while they readily take part in electron transfer reactions with mediators in solution. Advantage was taken of the latter fact to develop a new method to study the kinetics and equilibria of protein-mediator electron transfer reactions. It was shown that rate and equilibrium constants for the electron exchange between electron transfer mediator and the protein can be obtained from the analysis of the perturbation of the linear sweep voltammetry (LSV) response of the mediator due to the presence of the protein. The experiments were carried out under conditions where the protein does not interact with the electrode. Theoretical data obtained by digital simulation are presented to show the conditions under which rate and equilibrium constants are accessible by the LSV technique. The electron transfer reactions between ferri- and ferrocytochrome c and N,N,N',N'-tetramethylphenylenediamine and the corresponding radical cation in phosphate-buffered saline (0.04 M phosphate, pH 7.4, 0.1 M NaCl) buffer were selected to demonstrate the technique. These studies resulted in an equilibrium constant equal to 1.0 and forward and reverse rate constants equal to 1.6 x 10(4) M-1 s-1. The data available from this method include forward and reverse rate constants for electron transfer and the formal potential for the protein redox couple. PMID- 9212874 TI - Single-chain Fv fusion proteins suitable as coating and detecting reagents in a double antibody sandwich enzyme-linked immunosorbent assay. AB - A recombinant enzyme-linked immunosorbent assay (ELISA) system, entirely based on antibody fragments, is described here as an attractive alternative to assays using polyclonal antisera or monoclonal antibodies. Two expression vectors have been developed for cloning and production of single-chain Fv (scFv) fusion proteins suitable as coating and detecting reagents, respectively, in a highly sensitive double antibody sandwich ELISA. The coating reagent is produced from the vector pZIP1, as a bivalent miniantibody with a leucine zipper for dimerization. The detection reagent is a fusion protein, in which a modified Escherichia coli alkaline phosphatase with increased specific activity is attached to the carboxy-terminus of the scFv. This conjugate is produced using pDAP2/S as cloning and expression vector. Optimized bacteria expression and simple one-step purification by hexahistidine tag-mediated metal chelate affinity chromatography yielded milligrams of ELISA reagent per liter of bacterial culture in both cases. A double antibody sandwich ELISA for the detection of beet necrotic yellow vein virus, the causal agent of sugarbeet rhizomania, was developed using fusion proteins obtained by means of pZIP1 and pDAP2/S. The plant pathogen was detected with a sensitivity higher than that reached in a conventional ELISA employing polyclonal antisera. Both plasmid vectors are compatible to phage display vectors such as pHEN1, pCOCK, and the pCANTAB series, allowing simple subcloning after isolation of scFv from phage display libraries. It is therefore easy to develop and produce an ELISA entirely by using bacterial recombination and expression techniques. PMID- 9212875 TI - Characterization of PicoGreen reagent and development of a fluorescence-based solution assay for double-stranded DNA quantitation. AB - A sensitive assay for detecting double-stranded (ds) DNA in solution is described. This assay employs a new dye, PicoGreen dsDNA quantitation reagent, which becomes intensely fluorescent upon binding nucleic acids. The brightness of this reagent is due to its high quantum yield (approximately 0.5, bound to ds calf thymus DNA) and large molar extinction coefficient (approximately 70,000 cm 1 M-1). The fluorescence enhancement of this dye upon binding dsDNA is > 1000 fold, with excitation and emission maxima near those of fluorescein. Unlike Hoechst 33258, PicoGreen reagent fluorescence intensity was the same upon binding to poly(dA).poly(dT) and poly(dG).poly(dC) homopolymers. The PicoGreen assay allowed the detection of 25 pg/ml dsDNA, surpassing the sensitivity achieved with Hoechst 33258 by 400-fold. The linear concentration range for DNA quantitation extended over four orders of magnitude-25 pg/ml to 1 microgram/ml-with a single dye concentration. Assay linearity was maintained even in the presence of salts, proteins, poly(ethylene glycol), urea, chloroform, ethanol, and agarose; some ionic detergents and heparin interfered. Linear DNAs yielded slightly brighter signals than supercoiled plasmids. Finally, the assay showed greater dsDNA:RNA selectivity than Hoechst 33258 in low ionic strength buffer and better dsDNA:single-stranded DNA selectivity in 1 M NaCl. PMID- 9212876 TI - An inverse mammalian two-hybrid system for beta secretase and other proteases. PMID- 9212878 TI - Evaluation of antioxidant capacity by chemiluminescence. PMID- 9212879 TI - Isolation of cytoplasmic RNA from small samples of whole blood. PMID- 9212877 TI - Quantitative determination of 2-monovinyl protochlorophyll(ide) b by spectrofluorometry. PMID- 9212880 TI - A method for the measurement of plasma estrone fatty ester levels. PMID- 9212881 TI - A rapid method for the preparation of yeast lysates that facilitates the immunodetection of proteins generated by the yeast two-hybrid system. PMID- 9212882 TI - Detection of calcium binding proteins by two-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis. PMID- 9212884 TI - Topics in pulmonary nuclear medicine. PMID- 9212883 TI - Recent advances in nuclear cardiology in the study of coronary artery disease. AB - A variety of new radiopharmaceutical agents have been introduced to probe myocardial function in vivo. This review will introduce these new techniques which have recently been available in Japan. Tc-99m perfusion imaging agents provide excellent myocardial perfusion images which may enhance diagnostic accuracy in the study of coronary artery disease. In addition, greater photon flux from the tracer permits simultaneous assessment of regional perfusion and function with use of first-pass angiography or ECG-gated acquisition. Positron emission tomography enables metabolic assessment in vivo. Preserved FDG uptake indicates ischemic but viable myocardium which is likely to improve regional dysfunction after revascularization. In addition, FDG-PET seems to be valuable for selecting a high risk subgroup. Recently I-123 BMIPP, a branched fatty acid analog, has been clinically available in Japan. Less uptake of BMIPP than thallium is often observed in the ischemic myocardium. Such perfusion metabolic mismatch which seems to be similarly observed in FDG-PET is identified in the stunned or hibernating myocardium with regional dysfunction. Both of them are likely to recover afterwards. Severe ischemia is identified as reduced BMIPP uptake at rest, suggesting its role as an ischemic memory imaging. I-123 MIBG uptake in the myocardium reflects adrenergic neuronal function in vivo. In the study of coronary artery disease, neuronal denervation is often observed around the infarcted myocardium and post ischemic region as well. More importantly, reduced MIBG uptake in these patients can identify high risk for ventricular arrhythmias and assess severity of congestive heart failure. These new techniques will provide insights into new pathological states in the ischemic heart disease and enable to select optimal treatment in these patients. PMID- 9212885 TI - Diagnosis of chronic liver disease from liver scintiscans by artificial neural networks. AB - Artificial neural networks were used in the diagnosis of chronic liver disease based on liver scintiscanning. One hundred and thirty-seven patients with chronic liver disease (12 with chronic persistent hepatitis, 39 with chronic aggressive hepatitis, and 86 with cirrhosis) and 25 healthy controls were studied. Sixty five subjects (10 healthy controls, 20 patients with chronic hepatitis, and 35 patients with cirrhosis of the liver) were used in the establishment of a neural network. Liver scintiscans were taken starting 20 min after the intravenous injection of 111 MBq of Tc-99m-phytate. The neural network was used to evaluate five items judged from information on liver scintiscans: the ratio of the sizes of the left and right lobes, splenomegaly, radioactivity in the bone marrow, deformity of the liver and distribution of radioactivity in the liver. The neural network was designed to distinguish between three liver conditions (healthy liver, chronic hepatitis and cirrhosis) on the basis of these five items. The diagnostic accuracy with the neural network was 86% for patients with chronic hepatitis and 93% for patients with cirrhosis. With conventional scoring, the accuracy was 77% for patients with chronic hepatitis and 87% for patients with cirrhosis. Our findings suggest that artificial neural networks may be useful for the diagnosis of chronic liver diseases from liver scintiscans. PMID- 9212886 TI - Binding of a human monoclonal antithyroglobulin antibody to cultured human thyroid cancer cells. AB - To develop a new method of radioimmunodetection for thyroid cancer, we tested the binding ability of a human antithyroglobulin monoclonal antibody, VB5, to primary culture of human thyroid cancer cells. VB5 was able to immunostain cytoplasmic thyroglobulin (Tg) in the acetone-fixed cancer cells when used in a labeled streptavidin-biotin method but not in a conventional indirect immunoperoxidase technique. The antibody was readily labeled with I-125 in the standard chloramin T method, and showed specific binding to the antigen on cultured malignant thyrocytes displaceable with non-labeled VB5 or with excess Tg antigen. Although these initial results in vitro are encouraging, the observed low specific binding (about 1% at room temperature) to intact cells with a single monoclonal antibody seems insufficient to conduct any in vivo immunolocalization experiments in animals. To obtain more binding, we would need a cocktail of several monoclonal antibodies to different epitopes, and also fragmentation of antibody molecules to penetrate into cytoplasm. PMID- 9212887 TI - Functional imaging of gated Tc-99m tetrofosmin study as a simple method to quantify ventricular wall motion. AB - Myocardial perfusion scintigraphy with wall motion analysis is known to enhance accuracy in diagnosing ischemic heart disease. The purpose of this study is to determine the best method to evaluate regional wall motion in a gated planar perfusion study. Planar gated 99mTc tetrofosmin (GTF) study in two projections was performed after rest-exercise sequence SPECT studies (n = 29). To evaluate wall motion, cine-mode display, wall thickening, and inverted tetrofosmin studies including ventricular inner border tracing, segmental wall shortening and functional images were used. The results were compared with gated blood-pool (GBP) study in the same projections. In the GTF study, functional image identified asynergy significantly better than cinematic display. The best correlation between GTF and GBP studies was observed with functional images of phase and amplitude, with complete visual agreement seen in 145 of 168 (86%) segments. With quantitative analysis by means of regions of interest (n = 280), a good correlation was observed between GTF and GBP regarding regional amplitude (r = 0.78), regional phase (r = 0.84), average left ventricular phase (r = 0.91) and standard deviation of phase values (r = 0.90). The value for the count-based "ejection fraction" derived from inverted GTF showed insufficient correlation to that of the GBP study (r = 0.69). Functional imaging with myocardial perfusion imaging is a simple and effective means to evaluate ventricular asynergy. Similar diagnostic criteria to gated blood-pool imaging and comparable diagnostic accuracy are advantages of this approach. PMID- 9212888 TI - Accumulation of 99mTc-HMPAO and 99mTc-ECD in rodent and human breast tumor cell lines in vitro. AB - The accumulation of 99mTc-HMPAO and 99mTc-ECD was studied in rat (MatB) and human (MCF-7) breast tumor cell lines in vitro as a function of incubation time. The general pattern was the same for both tracers and both cell lines: the tracer rapidly and extensively accumulated in the cells but a plateau was reached in 15 30 minutes. Accumulation of HMPAO was higher than that of ECD, did not show a difference between rat and human cells, and correction of HMPAO data for intracellular sequestration and extracellular metabolism resulted in a linear increase in accumulation with time. In contrast, accumulation of ECD was approximately 2-fold higher in human cells than in rat cells but after correction for sequestration and metabolism a plateau remained. These experiments show differences between HMPAO and ECD in their accumulation and retention in breast cancer cells in vitro and support that the need for further work on the potential clinical role for HMPAO in tumor characterization. PMID- 9212889 TI - A method for the quantification of benzodiazepine receptors by using 123I iomazenil and SPECT with one scan and one blood sampling. AB - Iodine-123-iomazenil (Iomazenil) is a ligand of central type benzodiazepine receptors for single photon emission computed tomography (SPECT). Previously we reported a simple, table look-up method for quantification of its binding potential (BP) by using two SPECT scans and calibrated standard input function with one blood sampling. This method is based on a two-compartment model (K1: influx rate constant; k2: efflux rate constant; Vd (= K1/k2): the total distribution volume corresponding BP), and requires two SPECT scans for calculating both K1 and Vd values. If the K1 value in the two-compartment model can be assumed to be constant, the radioactivity of one SPECT scan at 180 min after injection can be considered to tabulate as a function of Vd for a given K1 value and a given input function, and a table look-up procedure provides the corresponding Vd value. The purpose of this study was to develop a simple, autoradiographic method for quantification of BP by using one SPECT scan and calibrated standard input function with one blood sampling. SPECT studies were performed on 14 patients. A dynamic SPECT scan was initiated following an intravenous bolus injection of Iomazenil. A static SPECT scan was performed at 180 min after the injection. Frequent blood sampling from the brachial artery was performed on all subjects to determine the arterial input function. Simulation studies revealed that errors in calculated Vd values were around +/-10-15% for varied K1 values. A good correlation was observed between total distribution volume values calculated by three-compartment model analysis and those calculated by the present method (r = 0.90), supporting the validity of this method. The present method is simple and applicable for clinical use, and will be able to provide images of BP. PMID- 9212890 TI - Assessment of technetium-99m Technegas scintigraphy for ventilatory impairment in pulmonary emphysema: comparison of planar and SPECT images. AB - Pulmonary emphysema can be diagnosed easily by X-ray CT (CT) as a low attenuation area. Recently Tc-99m-Technegas (Technegas) has been used for ventilation scintigraphy. The present study was undertaken to assess the usefulness of planar and SPECT images by using Technegas scintigraphy in patients with pulmonary emphysema. Technegas scintigraphy, CT and pulmonary function tests were performed in 20 patients (males, age 32-78 years). We classified the findings of Technegas images into 4 grades. Comparing planar and SPECT images of Technegas, more detailed findings were shown by SPECT than by planar images in mild cases (6 cases, 30%). In more severe cases, findings of SPECT and planar images were equivalent (14 cases, 70%). The degree of abnormal findings obtained by SPECT was equivalent to that obtained by CT in severe cases (6 cases, 30%). SPECT should be excluded in advanced stages as indicated by planar images. PMID- 9212891 TI - Carbon-11 labeled ethionine and propionine as tumor detecting agents. AB - To develop 18F-fluoroalkyl derivatives of methionine (MET) as a tumor detecting agent by mean of clinical PET, a pilot study assessing the potential of their parent compounds, 11C-labeled ethionine (11C-ETH) and propionine (11C-PRO), was performed. 11C-ETH and 11C-PRO were prepared by the reaction of L-homocysteine thiolactone and corresponding 11C-alkyl iodides. After i.v. injection of a mixture of 3H-MET. 14C-ETH and 11C-PRO into mice bearing FM3A mammary carcinoma, the highest FM3A uptake was found in 14C-ETH, followed by 3H-MET and 11C-PRO, while the FM3A-to-brain and FM3A-to-muscle ratios were nearly the same for all three compounds. The FM3A uptake of 14C-ETH and 11C-PRO were nearly equal or slightly higher than the liver uptake. In the pancreas, liver, FM3A and brain tissues, incorporation of 14C-ETH into acid-precipitable materials was much lower than that of 3H-MET, whereas no incorporation of 11C-PRO was found. Brain uptake of all three compounds was significantly reduced by carrier MET-loading (5 min p.i.) or by cycloheximide treatment to inhibit protein synthesis (60 min p.i.), whereas the FM3A uptake was not affected. Incorporation of 14C-ETH into acid precipitable materials was inhibited by the cycloheximide. The results suggest that 11C-labeled ETH has a similar potential for tumor detection by PET as 11C MET, and that 11C-PRO has similar properties to those of other artificial amino acids. The development of 18F-fluoroalkyl derivatives of MET is of interest as the next step. PMID- 9212892 TI - Precision of the gallbladder ejection fraction obtained with Tc-99m-pyridoxyl-5 methyl-tryptophan (99mTc-PMT) hepatobiliary scintigraphy as compared with the contraction ratio in three-dimensional computed tomography. AB - The gallbladder ejection fraction (GBEF) obtained with Tc-99m-pyridoxyl-5-methyl tryptophan (99mTc-PMT) hepatobiliary scintigraphy has been used as a parameter of gallbladder function. To determine the accuracy of GBEF, the relationship with the contraction ratio of the gallbladder (GBCR) obtained with three-dimensional helical computed tomography (3D-CT) was studied. PATIENTS AND METHODS: A normal volunteer, 8 patients suffering from cholecystolithiasis and a patient with gallbladder dyskinesia were examined. The percent initial dose (%ID) for the gallbladder and GBEF with hepatobiliary scintigraphy were used to compare the volume of the gallbladder and GBCR which was measured by 3D-CT. RESULTS: The %ID of the gallbladder was correlated with the volume of the gallbladder by 3D-CT (Y = 1.000X - 1.818, r = 0.928). GBEF was correlated well with GBCR by 3D-CT (Y = 0.916X + 6.296, r = 0.975). CONCLUSIONS: The %ID of the gallbladder obtained with hepatobiliary scintigraphy may be a good indicator of the volume of the gallbladder. The accuracy of GBEF was confirmed by comparison with 3D-CT examination. GBEF is considered a useful parameter of pathophysiological gallbladder function. PMID- 9212893 TI - Evaluation of left ventricular wall motion and function in patients with previous myocardial infarction by three-dimensional 99mTc-HSAD multigated cardiac pool imaging. AB - To evaluate left ventricular (LV) wall motion stereoscopically from all directions and to calculate the LV volume by three-dimensional (3D) imaging. 99mTc-DTPA human serum albumin-multigated cardiac pool-single photon emission computed tomography (99mTc-MUGA-SPECT) was performed. A new data processing program was developed with the Application Visualization System-Medical Viewer (AVS-MV) based on images obtained from 99mTc-MUGA-SPECT. In patients with previous myocardial infarction, LV function and LV wall motion were evaluated by 3D-99mTc-MUGA imaging. The LV end-diastolic volume (LVEDV) and end-systolic volume (LVESV) were obtained from 3D-99mTc-MUGA images by the surface rendering method, and the left ventricular ejection fraction (LVEF) was calculated at thresholds of 35% (T1), 40% (T2), 45% (T3), and 50% (T4). There was a strong correlation between the LV volume calculated by 3D-99mTc-MUGA imaging at a threshold of 40% and that determined by contrast left ventriculography (LVEDV: 194.7 +/- 36.0 ml vs. 198.7 +/- 39.1 ml, r = 0.791, p < 0.001; LVESV: 91.6 +/- 44.5 ml vs. 93.3 +/- 41.3 ml, r = 0.953, p < 0.001), respectively. When compared with the LVEF data obtained by left ventriculography, significant correlations were found for 3D images reconstructed at each threshold (T1: r = 0.966; T2: r = 0.962; T3: r = 0.958; and T4: r = 0.955). In addition, when LV wall motion obtained by 3D-99mTc-MUGA imaging (LAT and LAO views) was compared with the results obtained by left ventriculography (RAO and LAO views), there was good agreement. 3D-99mTc-MUGA imaging was superior in allowing evaluation of LV wall motion in all directions and in assessment of LV function, since data acquisition and image reconstruction could be done within a short time with the three detector imaging system and AVS-MV. This method appears to be very useful for the observation of both LV wall motion and LV function in patients with ischemic heart disease, because it is a noninvasive examination. PMID- 9212894 TI - Scan findings of various myocardial SPECT agents in a case of amyloid polyneuropathy with suspected myocardial involvement. AB - A 31-year-old male having familial amyloid polyneuropathy underwent a Tc-99m(V) DMSA study to evaluate the myocardial involvement. The patient also underwent T1 201, I-123-BMIPP and I-123-MIBG myocardial SPECT studies to evaluate blood perfusion, fatty acid metabolism and sympathetic function of the heart, respectively. Tc-99m(V)-DMSA SPECT showed uptake to the myocardium indicating myocardial involvement of amyloidosis. Both T1-201 and I-123-BMIPP studies showed normal uptake indicating normal blood perfusion and fatty acid metabolism but I 123-MIBG SPECT showed no uptake to the heart, indicating severe impairment of sympathetic function. PMID- 9212895 TI - Scintigraphic visualization of ocular melanoma with Tc-99m glutathione. AB - In a patient with ocular melanoma scintigraphy obtained with 99mTc-GSH clearly demonstrated the histologically proven ocular lesion both in planar and SPECT images. 99mTc-sestamibi study obtained in the same patient three days later was negative. 99mTc-GSH is a potential alternative to the currently used radiopharmaceuticals for imaging both cutaneous and ocular melanomas and their metastases. PMID- 9212897 TI - 111In-chloride uptake in pulmonary aspergillosis. AB - Unusual pulmonary uptake of 111In-chloride (111InCl3) was recognized in a patient with aplastic anemia. A chest radiograph showed infiltrative shadow concordant with 111InCl3 uptake, and autopsy revealed pulmonary aspergillosis. To our knowledge, there is not an extensive literature on 111InCl3 uptake in pulmonary aspergillosis. PMID- 9212896 TI - Visualization of uveal amelanotic melanoma with technetium-99m(V) dimercaptosuccinic acid. AB - The possibility of using technetium-99m(V) dimercaptosuccinic acid, Tc-99m DMSA, in the evaluation of uveal amelanotic melanoma was assessed in this study. Both planar and SPECT images clearly demonstrated the tumor. Following confirmation of our results by contemporaneous ultrasonography and MRI the patient was treated with Iodine-125 brachytherapy. In combination with other diagnostic tests, Tc 99m(V) DMSA scintigraphy may play a role in the detection of uveal melanoma and its possible systemic metastases. PMID- 9212898 TI - Acute gastric dilatation: an incidental finding in a Meckel study. PMID- 9212899 TI - Laurence-Moon-Biedl syndrome: scintigraphic appearance of kidneys. AB - We report a 7-year-old child with Laurence-Moon-Biedl syndrome, an autosomal recessive syndrome, with impaired renal function detected by means of technetium 99m diethylenetriamine-pentaacetic acid (Tc-99m DTPA), technetium-99m dimercaptosuccinic acid (Tc-99m DMSA) scintigraphy, and ultrasonography. The altered renal morphology and decreased renal functions are documented. PMID- 9212900 TI - Inferior mesenteric varix demonstrated by 99mTc-red blood cell gastrointestinal bleeding study. AB - Reported here is a case of an inferior mesenteric varix demonstrated by 99mTc-red blood cell (RBC) scintigraphy performed for gastrointestinal bleeding in a 47 year-old man. It was shaped like a question mark ranging from the left upper abdomen to the pelvis. This is the first report of scintigraphic recognition of an inferior mesenteric varix. PMID- 9212901 TI - Cerebral perfusion changes in traumatic diffuse brain injury; IMP SPECT studies. AB - Diffuse brain injury (DBI) is characterized by axonal degeneration and neuronal damage which cause diffuse brain atrophy. We have investigated the time course of abnormalities in cerebral perfusion distribution in cases of DBI by using Iodine 123-IMP SPECT, and the relationship to the appearance of diffuse brain atrophy. SPECT scans were performed on eight patients with diffuse brain injury due to closed cranial trauma in acute and chronic stages. All patients showed abnormalities in cerebral perfusion with decreases in perfusion, even in non depicted regions on MRI, and the affected areas varied throughout the period of observation. Diffuse brain atrophy appeared in all patients. In some patients, diffuse brain atrophy was observed at or just after the time when the maximum number of lesions on SPECT were seen. The abnormalities in cerebral perfusion in cases of DBI might therefore be related to axonal degeneration and neuronal damage which causes diffuse brain atrophy. PMID- 9212902 TI - Performance study of a miniature gamma ray scintillation vivo probe for tumor localization. AB - We have developed a miniature gamma-ray endoscopic probe consisting of dual BGO detector probes for tumor detection inside the body cavities. The dual detector system was coupled with random coincidence to decrease the distant background radiation and to improve its spatial resolution for tumor localization. METHOD: The performance of the probe was investigated with a point source and a water phantom. A solution of positron emitting 18F isotope was used as the source. Clinical trials of the probe were done to localize tumors on the skin surface of four subjects carrying tumors close to the body surfaces, into whom 67Ga-citrate and 18F-FDG radiopharmaceuticals were injected. RESULTS: Measurements indicated that the spatial resolution of the dual detector probes is around 1.5 times better than the single detector probe, and both single and dual detector endoscopic probe systems are capable of localizing a tumor on a large photon background. CONCLUSION: The endoscopic probe may be easier to insert inside body cavities due to the small crystal size and the flexible light guides. A single detector probe with higher sensitivity may be useful in searching for tumors over a wide intracavity area but a dual detector probe can be used for precise tumor localization. The detector probe may also be suitable for intraoperative observation. PMID- 9212903 TI - Rapid measurement of modified oligonucleotide levels in plasma samples with a fluorophore specific for single-stranded DNA. AB - Animal studies of therapeutic oligonucleotides require measurement of circulating levels of oligonucleotides by multistep, time-consuming methods. In contrast, addition of a single-stranded DNA binding fluorophore, OliGreen, to oligonucleotides in plasma samples allowed rapid quantitation. Dose-response curves were measured for five different oligonucleotide analogs added to plasma or serum. Phosphorothioate or 3'-amino phosphodiester oligodeoxynucleotides in calf serum reliably exhibited linear, dose-dependent fluorescence at 15-500 nM. The assay was equally sensitive in human and mouse plasma, with a heterogeneous variety of sequences. Oligonucleotides shorter than 10 nucleotides yielded substantially reduced fluorescence. In contrast, 2'-O-methyl oligoribonucleotides, DNA methylphosphonates, and peptide nucleic acids demonstrated little or no fluorescence with OliGreen. Following intravenous injection of a phosphorothioate pentadecamer into mice, fluorescence measurements of plasma phosphorothioate levels displayed a dose-dependent, biexponential decline over a 90 min period. Chronic infusion at 2.5 nmol/hour into mice yielded plasma oligonucleotide values equivalent to 0.1 microM, a value reflecting the contributions of intact and partially degraded strands. Tumor-bearing mouse plasma evidenced high fluorescence values in the absence of oligonucleotide administration, presumably because of elevated intrinsic plasma DNA fragments. Although limited in its ability to differentiate intact from partially degraded strands, OliGreen fluorescence provides a simple, rapid, and sensitive method for measuring circulating levels of phosphorothioate or phosphodiester oligonucleotides in healthy animals or humans. PMID- 9212904 TI - Tissue distribution and metabolism of the [32P]-labeled oligodeoxynucleoside methylphosphonate-neoglycopeptide conjugate, [YEE(ah-GalNAc)3]-SMCC-AET-pUmpT7, in the mouse. AB - Development of oligodeoxynucleotides (oligo-dNs) and their analogs as therapeutic agents is complicated by their low rate of transport across cellular membranes, which is required for interaction with the intracellular complementary nucleic acid sequences, and the lack of tissue-specific delivery. To overcome these obstacles, bioconjugates between cell surface receptor ligands and oligodeoxynucleoside methylphosphonates (oligo-MPs) have been constructed containing homogeneous, chemically defined covalent linkages. We have previously established that a model conjugate, [32P]-labeled [YEE(ah-GalNAc)3]-SMCC-AET pUmpT7 (1), is delivered to Hep G2 cells in a ligand-specific manner, reaching a peak value of 26 pmol per 10(6) cells after 24 hours incubation at 37 degrees C (Hangeland et al., 1995). In this work, the in vivo behavior of this conjugate is explored. Administration of this conjugate to mice via tail vein injection demonstrates rapid uptake in liver to the extent of 69.9 +/- 9.9% of the injected dose after 15 minutes. Thereafter, the conjugate and its metabolites are rapidly cleared via the kidney and urine. Polyacrylamide gel electrophoresis analysis of extracts of Hep G2 cells and mouse liver reveal the conjugate 1 to be extensively metabolized. In contrast, the conjugate found in mouse urine is largely intact. These data show that this novel, biodegradable delivery vehicle represents a viable approach for the delivery of antisense oligo-MPs and other oligo-dN analogs to the liver for therapeutic and diagnostic applications. PMID- 9212905 TI - A specificity comparison of four antisense types: morpholino, 2'-O-methyl RNA, DNA, and phosphorothioate DNA. AB - Cell-free translation studies were carried out to compare the efficacy and specificity of four antisense structural types: DNA, phosphorothioate DNA (S DNA), 2'-O-methyl RNA, and Morpholino oligos, a novel antisense structural type. In these studies, translational inhibition was assessed for two 20-mers of each structural type, where one 20-mer was complementary to its target sequence in rabbit alpha-globin mRNA and the other 20-mer contained three mispairs to that same target sequence. It is shown that at low concentration of antisense oligomer (50 nM) all four types provide high specificity, but the Morpholino oligos and 2' O-methyl RNA afford better efficacy. At high oligomer concentration (3.5 microM), all four types provide high efficacy, but the Morpholino oligos and 2'-O-methyl RNA provide substantially better specificity than the DNA and S-DNA. PMID- 9212906 TI - In vivo metabolic profile of a phosphorothioate oligodeoxyribonucleotide. AB - Antisense phosphorothioate oligodeoxyribonucleotides (PS oligonucleotides) have the ability to inhibit individual gene expression in the potential treatment of cancer and viral diseases. Following administration in vivo, PS oligonucleotides are rapidly cleared from the plasma and distributed to various organs. However, the manner in which administered oligonucleotides are metabolized in plasma and tissues is poorly understood. In this study, a 25-mer PS oligonucleotide (GEM91) complementary to the gag gene mRNA of the human immunodeficiency virus (HIV-1) was administered to mice through intravenous injections to investigate its metabolism. The PS oligonucleotide was extracted from plasma at 1 hour postadministration and from kidney and liver at 24 hours postadministration. After extraction, the PS oligonucleotide and its metabolites were tailed with dA and annealed to a dT-tailed plasmid. The recombinant plasmid was ligated and used to transform competent bacteria. The region of interest containing the PS oligonucleotide was then sequenced. Our results show that degradation of the PS oligonucleotide in plasma was primarily from the 3'-end. However, in kidney and liver, degradation was primarily from the 3'-end, but a large proportion of the PS oligonucleotide was degraded from the 5'-end as well. We also studied the metabolism of PS oligonucleotide in plasma after 2-hour intravenous infusion in HIV-infected patients. The degradation of the PS oligonucleotide in plasma was primarily from the 3'-end. This study is important in understanding the metabolism of antisense PS oligonucleotide in vivo in general but also provides guidance for designing second-generation antisense oligonucleotides with improved stability and safety profile. PMID- 9212907 TI - Identification of a phosphodiester hexanucleotide that inhibits HIV-1 infection in vitro on covalent linkage of its 5'-end with a dimethoxytrityl residue. AB - It has been shown in previous reports that a guanine-rich phosphodiester oligonucleotide bearing a dimethoxytrityl (DmTr) residue on its 5'-terminal. DmTr TGGGAGGTGGGTCTG (SA-1042), is an inhibitor of HIV-1 infection in vitro. SA-1042 interfered with the attachment of gp120 to the CD4 receptor and the subsequent entry stage of viral infection. We investigated the structure-activity relationship of the DmTr-conjugated oligomer by using 15-mer oligonucleotides with various nucleotide sequences. Results show that location of guanine nucleosides at the 5'-terminal and modification of the 5'-terminal with DmTr are essential for anti-HIV-1 activity. First, substitution of the guanine nucleoside close to the 5'-terminal of SA-1042 with another nucleotide prevented antiviral activity. Second, the existence of at least three consecutive guanine nucleosides adjacent to the 5'-terminal was required for the activity. Finally, modification of the 5'-terminal was essential for the activity. Based on these findings, the hexanucleotide, DmTr-TGGGAG, was identified as a potent inhibitor of HIV-1 infection. The hexamer was found to be capable of inhibiting the binding of gp120 to its receptor CD4 molecule, and it was also capable of inhibiting accessibility of anti-V3 monoclonal antibody to its ligand V3 peptide. PMID- 9212909 TI - Morpholino antisense oligomers: design, preparation, and properties. AB - Antisense promised major advances in treating a broad range of intractable diseases, but in recent years progress has been stymied by technical problems, most notably inadequate specificity, ineffective delivery into the proper subcellular compartment, and unpredictable activity within cells. Herein is an overview of the design, preparation, and properties of Morpholino oligos, a novel antisense structural type that solves the sequence specificity problem and provides high and predictable activity in cells. Morpholino oligos also exhibit little or no nonantisense activity, afford good water solubility, are immune to nucleases, and are designed to have low production costs. PMID- 9212908 TI - Delivery of oligoribonucleotides to human hepatoma cells using cationic lipid particles conjugated to ferric protoporphyrin IX (heme). AB - The receptor-ligand interaction between hepatocyte heme receptors and heme was evaluated as a basis for developing a targeted cationic lipid delivery reagent for nucleic acids. Heme (ferric protoporphyrin IX) was conjugated to the aminolipid dioleoyl phosphatidylethanolamine (DOPE) and used to form cationic lipid particles with dioleoyl trimethylammonium propane (DOTAP). These lipids particles (DDH) protect oligoribonucleotides from degradation in human serum and increase oligoribonucleotide uptake into 2.2.15 human hepatoma cells (to a level of 50-60 ng oligo/10(4) cells) when compared with the same lipid particles (DD) prepared identically without heme. The DDH heme level that was optimal for oligoribonucleotide delivery was also optimal for maximum expression of plasmid encoded luciferase. The enhancing effect of heme was evident only at net particle negative charge. Fluorescence microscopy showed that DDH delivered oligoribonucleotides into both the 2.2.15 cell cytoplasm and nucleus. DDH may thus be a potentially useful delivery vehicle for oligonucleotide-based therapeutics and transgenes, appropriate for use in such liver diseases as viral hepatitis, hepatoma, and hypercholesterolemia. PMID- 9212910 TI - Background of the antisense oligonucleotide approach to chemotherapy. PMID- 9212911 TI - Recruiting the 2-5A system for antisense therapeutics. PMID- 9212912 TI - Controversies in the cellular pharmacology of oligodeoxynucleotides. PMID- 9212913 TI - Recombinational repair of genetic mutations. PMID- 9212914 TI - Antisense-protein kinase A: a single-gene-based therapeutic approach. PMID- 9212915 TI - Antisense c-myc inhibition of lymphoma growth. PMID- 9212916 TI - Identification and characterization of second-generation antisense oligonucleotides. PMID- 9212917 TI - Protein kinase C as a target for cancer therapy. PMID- 9212918 TI - Pharmacology of therapeutic oligonucleotides. PMID- 9212919 TI - In vivo pharmacokinetics of phosphorothioate oligonucleotides containing contiguous guanosines. PMID- 9212920 TI - Ex vivo bone marrow purging with oligonucleotides. PMID- 9212921 TI - Antisense transforming growth factor-beta 1 in wound healing. PMID- 9212922 TI - Rotary blood pump: paracorporeal, implantable, percutaneous? PMID- 9212923 TI - Therapeutic and physiological artificial heart: future prospects. AB - Current left ventricular assist devices (LVADs) have demonstrated admirable results. However, approximately one-fourth of the patients who require LVADs suffer from right heart failure and require additional right ventricular (RV) assist devices (RVADs). The RV failure impairs the splanchnic circulation, subsequently developing into multiorgan failure (MOF). An aggressive application of a biventricular assist device (BVAD) is the best way to avoid and treat MOF because the BVAD reduces splanchnic congestion. Also, because the BVAD allows retention of the natural heart, recovery of the heart function can be expected after long-term assist. This benefit cannot be expected from conventional total artificial hearts. Although there are no implantable clinical BVAD systems in existence today, present advanced technologies in rotary blood pumps can enable these systems to be totally implantable. So, we should focus on developing a totally implantable BVAD system. The implantable BVAD will be a therapeutic and physiological total artificial heart, and it will be a common home health care device in the near future. PMID- 9212924 TI - Development and initial testing of a permanently implantable centrifugal pump. AB - To be able to salvage heart failure patients, the need for an economical permanent ventricular assist device is increasing. To meet this increasing demand, a miniaturized centrifugal blood pump has been developed as a permanently implantable device. The Gyro permanently implantable model (PI-601) incorporates a sealless design with a blood stagnation free structure. The pump impeller is magnetically coupled to the driver magnet in a sealless manner. This pump is atraumatic and antithrombogenic and incorporates a double pivot bearing system. A miniaturized actuator was utilized in this system in collaboration with the University of Vienna. The priming volume of this pump is 20 ml. The overall size of the pump actuator package is 53 mm in height and 65 mm in diameter, 145 ml of displacement volume, and 305 g in weight. Testing to date has included in vitro hydraulic performance and hemolysis. This pump can provide 5 L/min against a 110 mm Hg total pressure head at 2,000 rpm and 8 L/min against 150 mm Hg at 2,500 rpm. The normalized index of hemolysis (NIH) value of this pump was 0.0028 g/100 L at 5 L/min against 100 mm Hg. A preliminary anatomical study revealed the possibility of the implantability of 2 such systems in biventricular bypass at a preperitoneal location. This system is feasible for use as a permanently implantable biventricular assist device. PMID- 9212925 TI - Development of the Nimbus/Pittsburgh axial flow left ventricular assist system. AB - Nimbus, Inc. and the University of Pittsburgh's School of Medicine have been collaborators developing rotary blood pump technology since 1992. Currently, a major focus is on an implantable left ventricular assist system (LVAS) that utilizes an electric powered axial flow blood pump. In addition to the blood pump, a major development item is the electronic controller and the control algorithm for modulating the pump speed in response to varying physiologic demands. Methods being used in developing the axial flow LVAS include the use of computational fluid dynamic modeling of the interior flow field of the pump, flow visualization of the flow field using laser based imaging, and computer simulation of blood pump-physiological interactions as well as an extensive in vivo test program. Results to date include successful in vivo tests of blood pumps with nonlubricated bearings and demonstrations of auto speed control using electrical current as the observable parameter. PMID- 9212927 TI - New mechanism to reduce the size of the monopivot magnetic suspension blood pump: direct drive mechanism. AB - Size reduction of the monopivot magnetic suspension blood pump has been achieved by reducing the size of the magnetic suspension and employing a direct drive mechanism in place of a brushless DC motor and a magnetic coupling. The flow has also been improved using a closed hollow impeller to remove flow obstruction at the inlet and using radial straight vanes to reduce the impeller speed by 30%. Hemolysis testing was conducted for the new models. Results showed that model DD1 presented only a slightly higher level of hemolysis than a regular extracorporeal centrifugal pump. PMID- 9212926 TI - Purge system for rotary blood pumps. AB - Purge liquid has been supplied successfully to lubricate the bearings and wash the seals of rotary blood pumps to minimize hemolysis and thrombosis, extending their operating endurance. Although encouraging results have been obtained and development is proceeding for pumps with blood lubricated bearings, their validation and clinical use will occur some time in the future. For rotary blood pumps with purged bearings, a miniature purge liquid pump weighing 80 g has been successfully developed to meet the design point of 1 ml/h at 1,000 mm Hg (required by an existing rotary blood pump) with a motor power of 4 mW. The novel device maintains the flow rate for a sufficient time to service or replace the pump without flow interruption. A preliminary design has been made for a wearable purge delivery unit comprising a purge liquid pump, an electronic control, a purge liquid reservoir, and a battery. PMID- 9212928 TI - Nonlinear mathematical analysis of the hemodynamic parameters during left ventricular assistance with oscillated blood flow. AB - For the development of a totally implantable ventricular assist system (VAS), we have been developing the vibrating flow pump (VFP), which can generate oscillated blood flow with a relatively high frequency (10-50 Hz) for a totally implantable system. In this study, effects of left ventricular assistance with this unique oscillated blood flow were analyzed by nonlinear mathematics for evaluation as the entire circulatory regulatory system, not as a separate part of the system. Left heart bypasses using VFPs from the left atriums to the descending aortas were performed in chronic animal experiments using healthy adult goats. Electrocardiogram (ECG), arterial blood pressure, VFP pump flow, and flow of the descending aorta data taken while the goats were awake were recorded in the data recorder and analyzed in the personal computer system through the AD convertor. Using nonlinear mathematics, time series data were embedded into the phase space, and the Lyapunov numerical method, fractal dimension analysis, and power spectrum analysis were performed to evaluate the nonlinear dynamics. During left ventricular assistance with the VFP, Mayer wave fluctuations were decreased in the power spectrum, the fractal dimension of the hemodynamics was significantly decreased, and peripheral vascular resistance was significantly decreased. These results suggest that nonlinear dynamics, which mediate the cardiovascular dynamics, may be affected during LV bypass with oscillated flow. Decreased power of the Mayer wave in the spectrum caused the limit cycle attractor of the hemodynamics and decreased the peripheral resistance. Decreased sympathetic discharges may be the origin of the decreased Mayer wave and fractal dimension. These nonlinear dynamical analyses may be useful to design the optimal VAS control. PMID- 9212930 TI - Chronic animal experiment with magnetically suspended centrifugal pump. AB - We have been developing a new type of centrifugal pump for long-term use. The magnetically suspended centrifugal pump (MSCP) contains no shaft and seal so that long life expectancy is predicted. Paracorporeal left ventricular (LV) assist circulation between the left atrium and the descending aorta was instituted using sheep. The flow rates ranged from 2.5-5.5 L/min. The sheep that lived the longest (46 days) died of an embolism as a result of the thrombus in the pump. No thrombus formation was observed in other pumps. Plasma free hemoglobin levels ranged from 9 to 18 mg/dl, which led to the conclusion that the hemolysis level remained within an acceptable range. Two driving modes were compared. The slope of the pressure-flow relationship plot under a constant motor current mode was steeper than that under a constant rotational speed mode, and thus, the flow fluctuation decreased. In conclusion, the MSCP is durable for more than a month at the current stage of development and is a promising device for long-term ventricular assist. PMID- 9212929 TI - An automatic flow controller for a centrifugal blood pump. AB - To regulate the perfusion flow rate of a centrifugal blood pump, a microcomputer controller was developed. The computer monitored the flow rate of the pump with an electromagnetic flowmeter or an ultrasonic pulse Doppler flowmeter, rotational speed of the pump, aortic pressure, and the amount of blood in a reservoir. A discrete integral controller with a control interval of 1 s was adopted for the controller. For the safety of the control system, we added functions for detecting a clamp on the tubing, a dislocation of the flow sensor, or an inverse direction of the flow sensor. During a standby period, the computer calculated the rotational speed from aortic pressure to minimize the forward or the backward flow at the start of the pump perfusion. The automatic flow controller was used on 5 patients during cardiac operations and maintained the flow rate within +/-6% of the set point. PMID- 9212931 TI - Long-term evaluation of a nonpulsatile mechanical circulatory support system. AB - Antithrombogenicity of a centrifugal pump (CP) developed in our institute is provided by a central balancing hole (BH) in the impeller. A current CP, the National Cardiovascular Center (NCVC)-2, was ameliorated to improve antithrombogenicity, whereby the BH diameter was widened to improve self washout flow velocity, and an edge of the thrust bearing was rounded off to minimize flow separation. Effects of these modifications were assessed in a long-term in vivo experiment. The antithrombogenicity, hemolytic property, and mechanical durability of the NCVC-2 were investigated in 3 goats. The NCVC-2 was installed paracorporeally between the left atrium and the aorta and driven as long as possible at rotating speeds of about 2,800 rpm. The NCVC-2 ran for 50, 200, and 367+ days. The mean bypass flow rates were 6.8, 5.0, and 5.3 L/min, respectively. Creatinine, blood urea nitrogen (BUN), glutamic-oxaloacetic transaminase (GOT), and glutamic-pyruvic transaminase (GPT) did not increase until one week before termination. Plasma free hemoglobin was kept to a level less than 15 mg/dl, except for the last week of the second case. These results indicate that the NCVC 2 has excellent antithrombogenicity, an acceptable hemolytic property and the necessary durability for prolonged use. PMID- 9212932 TI - Design considerations for bearingless rotary pumps. AB - The designs of rotary blood pumps have shown substantial technical progress over recent years, especially contact bearing designs. However, the concern for potential thromboembolism remains and can only be eliminated by the use of bearingless pumps. Bearingless designs can be achieved through the application of magnetic, hydrodynamic, and hydrostatic forces or a proper combination of these forces. Although a purely magnetically suspended, actively controlled system can be designed, judicious use of hydraulic forces can allow simplification of device configuration and control. In this study, bearingless designs were evaluated for both axial and centrifugal pump configurations. Trade-offs between shear rates and bearing leak rates were considered based upon constraints imposed by hemolysis and residence time. These principles were used for determining the design feasibility of a rotary pump using combined magnetic and hydraulic stabilizing forces. PMID- 9212933 TI - Noninvasive pump flow estimation of a centrifugal blood pump. AB - A flow rate estimating method was investigated for a centrifugal blood pump developed in our institute. The estimated flow rate was determined by the power consumption, the rotating speed of the motor, and the hematocrit value. The power consumption and the rotating speed of the motor were measured with a wattmeter. The examinations were performed in a closed mock loop filled with goat blood with hematocrit values of 21.5%, 28%, 34%, and 42%. Measured values of blood viscosity were 2.47, 3.09, 3.71, and 5.07 mPa.s at a share rate of 37.5/s, respectively. A linear correlation between the power consumption and the pump flow rate was observed in all hematocrit values. But variations in hematocrit caused a difference in the flow rate up to 1.1 L/min at the same power consumption and rotating speed. Effects of blood viscosity on the flow estimation were corrected by the hematocrit value. The value of the coefficient of determination, R2, between the estimated flow rate and the measured flow rate was 0.988. These results may indicate that the flow estimating method calculated by the power consumption of the motor, the rotating speed, and the hematocrit value is useful in the clinical situation. PMID- 9212934 TI - Control of centrifugal blood pump based on the motor current. AB - In this study, centrifugal pump performance was examined in a mock circulatory loop to derive an automatic pump rotational speed (rpm) control method. The pivot bearing supported sealless centrifugal pump was placed in the left ventricular apex to aorta bypass mode. The pneumatic pulsatile ventricle was used to simulate the natural ventricle. To simulate the suction effect in the ventricle, a collapsible rubber tube was placed in the inflow port of the centrifugal pump in series with the apex of the simulated ventricle. Experimentally, the centrifugal pump speed (rpm) was gradually increased to simulate the suction effect. The pump flow through the centrifugal pump measured by an electromagnetic flowmeter, the aortic pressure, and the motor current were continuously digitized at 100 Hz and stored in a personal computer. The analysis of the cross-spectral density between the pump flow and motor current waveforms revealed that 2 waveforms were highly correlated at the frequency range between 0 and 4 Hz, with the coherence and phase angles being close to 1.0 and 0 degree, respectively. The fast Fourier transform analysis of the motor current indicated that the second harmonic component of the motor current power density increased with the occurrence of the suction effect in the circuit. The ratio of the fundamental to the second harmonic component decreased less than 1.3 as the suction effect developed in the circuit. It is possible to detect and prevent the suction effect of the centrifugal blood pump in the natural ventricle through analysis of the motor current waveform. PMID- 9212935 TI - Wireless monitoring and control for implantable rotary blood pumps. AB - A wireless biotelemetry system for the transfer of digital data through intact skin and tissue has been developed to provide a safe and noninvasive means of communication between implanted medical devices and the outside of the body. The system utilizes 2 miniature infrared transmitter/receiver modules. Data are transmitted through intact skin and subcutaneous tissue on an 890 nm infrared carrier signal. The system has been evaluated in human cadavers and during in vivo implantation of artificial hearts and ventricular assist devices for durations of up to 96 h. Acceptable data transfer (error rate < 10(-5)) through a typical tissue thickness of 5-25 mm has been demonstrated. The ability to monitor and control a device from a remote site using public communication systems such as telephone lines and asynchronous transfer mode (ATM) systems has also been demonstrated. Design optimization is currently ongoing in preparation for clinical utilization with artificial heart systems and other implantable devices (such as rotary blood pumps). PMID- 9212936 TI - Development of the undulation pump total artificial heart. AB - The undulation pump is a small size continuous flow displacement type blood pump that has been developed for an artificial heart. Using undulation pumps, 2 types of implantable total artificial hearts (TAHs), the undulation pump TAH (UPTAH) type 1 (UPTAH 1) and UPTAH type 2 (UPTAH 2) were developed. Both UPTAHs were designed to be small enough to implant into the chest of a goat, the experimental animal. UPTAH 1 could be reduced in size to 75 mm in diameter and 78 mm in length. The weight was 520 g. UPTAH 2 could be reduced in size to 75 mm in diameter and 80 mm in length. The weight was 650 g. UPTAH 2 could be tested in an animal experiment using an adult female goat weighing 52.3 kg. The UPTAH 2 could be implanted successfully into the goat's chest with a good fit. The goat stood after the surgery and extubation and survived for 3 h and 40 min; thus, the potential of the UPTAH for a practical implantable TAH was demonstrated. PMID- 9212937 TI - Realization of a permanent implantable pulsatile impeller heart with magnetically suspended motor. AB - A permanent impeller heart that could work for years was once an idea. However, now this idea is turning into reality through the use of the magnetically suspended motor. Recently, with our implantable pulsatile impeller pump, 3 left ventricular assisted calves survived for about 2 months (62, 54, and 46 days, respectively). The termination of the experiments was related to wear of the mechanical bearing, which resulted in vibration of the rotor and pump failure. All the experimental animals were in good condition prior to pump failure. It seemed as if the experiments could have lasted indefinitely if the bearing had not failed. All the hematological and biochemical data of the calves remained in normal or acceptable ranges; neither blood damage nor organ dysfunction of any animal was detected. During autopsy, no severe thrombus formation was found in the pump or vessels although a low dose of heparin (0.5-0.8 g/h) was given to increase the activated coagulation time (ACT) to 1.5-2.0 times its normal value. To solve the problem of bearing wear, a magnetically suspended motor was investigated and applied to the impeller pump. On the opposite sides of a disc connected to the rotor, 2 permanent magnet rings were embedded, one for driving and the other for axial suspension. Because both the driving and suspending coils with iron cores attract the disc, no radial bearing was needed. This newly devised impeller heart promises to have long-term and permanent applications. PMID- 9212938 TI - Flow visualization studies to improve the spiral pump design. AB - The spiral pump (SP) uses centrifugal and axial pumping principles simultaneously, through a conical shaped impeller with threads in its surface. Flow visualization studies were performed in critical areas of the SP. A closed circuit loop was filled with glycerin-water solution (40%). Amberlite particles (80 mesh) were illuminated by a planar helium-neon laser light (7 mW). The particle velocities were recorded with Kodak (TMAX-400) black and white film, and the flow behavior was studied with a micro video camera and color video printer. The flow visualization studies showed no turbulence or stagnant areas in the inlet and outlet ports of the SP. When using the impeller with one lead, at the top of the threads some recirculation appeared when the total pressure head increased. Two new impellers were made. One of them had the same conical shape with a thread having 2 leads. The second had a thread with 2 leads, but it also had a bigger cone angle. These modifications improved the pump hydrodynamic performance, decreasing the recirculation in pumping conditions that require pressures over 200 mm Hg. PMID- 9212939 TI - Hemolytic effect of surface roughness of an impeller in a centrifugal blood pump. AB - The present study investigates how the surface roughness of an impeller affects hemolysis in the pivot bearing supported Gyro C1E3 pump. This study focuses on particular areas of the impeller surface in the impeller type centrifugal pump. Seven Gyro C1E3 pumps were prepared with smooth surface housings and different impeller parts with different surface roughnesses. The vanes, top side, and backside of the impeller were independently subjected to vapor polishing, fine sand blasting, or coarse sand blasting to produce three different grades of surface roughness. These surfaces were then examined by a surface profile instrument. Using these pumps with different impellers, in vitro hemolysis tests were performed simulating cardiopulmonary bypass (5 L/min, 350 mm Hg). The findings of this study conclusively proved that surface roughness of the back side of the impeller has the greatest effect on hemolysis, followed by the top side and then the vanes. The following are reasons for these findings. First, the shear rate may be greater on the back side than on the top side because of the smaller gap between the back and the housing and the greater relative speed against the impeller. Second, the fluid beneath the impeller may have a longer exposure time because there is little chance for the fluid to mix beneath the impeller. Third, the shear rate may be greater on the top side of the impeller than on the vanes because a vortex formation occurs behind the vanes. PMID- 9212940 TI - Hemolysis test of disposable type vibrating flow pump. AB - The vibrating flow pump (VFP) can generate high frequency oscillated blood flow. Because of the high frequency driving with short stroke volume, the pump system can be small. The disposable type VFP (D-VFP) was developed for use for extracorporeal circulation. The electromagnetic actuator was detached from the vibrating tube, which was newly designed to be a disposable tube with a jellyfish valve. Hemolysis tests of the D-VFP, VFP, centrifugal pump, and roller pump were performed in a mock circulation study using goat blood. Plasma free hemoglobin was measured every 15 min under the same conditions. The plasma free hemoglobin of the D-VFP was 16 mg/dl although that of the VFP was 160 mg/dl at 30 min. The plasma free hemoglobin of the centrifugal pump and roller pump at 30 min were 3 mg/dl and 9 mg/dl, respectively. The hemolysis performance of the D-VFP may be studied further as a result of this study. Two important factors affecting hemolysis development may be the materials of which the vibrating tube is made and heat transmission from the actuator. The D-VFP has a smooth acrylic surface for blood contact compared with the metal surface of old type VFP. The electromagnetic actuator of the VFP surrounded the vibrating tube, so heat from the actuator could be easily transmitted to the blood. Because the D-VFP has a disposable vibrating tube that is detached from the actuator, heat is not readily transmitted to the blood. A mock circulation study of heat transmission was performed using the D-VFP and VFP. Results of the heat transmission study showed that the fluid temperature of the D-VFP was not increased and stayed at room temperature although that of the VFP increased approximately 1 degree C above room temperature. The D-VFP may be a good style for the development of the VFP for use for extracorporeal circulation. PMID- 9212941 TI - Hemolytic effect of the secondary vane incorporated into the back side of the impeller. AB - The hemolytic effect of the secondary vane system, the antithrombogenic structure incorporated into the back side of the impeller of the C1E3 Gyro pump, was investigated. Impellers with 0, 2, 3, and 4 secondary vanes and an additional impeller with 2 secondary channels were fabricated and incorporated into the C1E3 pump casings. Hemolysis tests were performed under cardiopulmonary bypass conditions (flow rate 4.5 L/min, total pressure head 350 mm Hg) using flesh bovine blood. The normalized indices of hemolysis (NIH) of the pumps with 0, 2, 3, and 4 secondary vanes and the pump with 2 secondary channels were 0.0797, 0.0866, 0.104, 0.157, and 0.0591, respectively. These results indicated that design of the impeller with 2 secondary channels, which was the original design of C1E3 Gyro pump, was less hemolytic than the design with secondary vanes. Additionally, the possibility of the secondary channel system for the impeller bottom was demonstrated favorably. PMID- 9212942 TI - Evaluation of hemolysis in a pulsatile assist device for centrifugal pump. AB - To evaluate the blood trauma caused by a new device for producing a pulsatile flow of the centrifugal pump, the pulsatile assist device for the centrifugal pump (PAD-CP) that we have developed, a hemolysis study was performed in vitro and in animal experimentation. For the in vitro testing, 2 identical sets of hemolysis test circuits were prepared with 2,400 ml of bovine blood. The 2 circuits were pumped simultaneously. Plasma total hemoglobin levels were less than 40 mg/dl after 3 h, under a pump flow of 2 L/min. Hemolysis increased to a severe level after 4 h of 4 L/min pump flow. The cause of this hemolysis was thought to be a vibration of the circuit because of incomplete compression of the polyurethane tube in the PAD-CP. Five adult sheep (average body weight, 47 kg) were used for in vivo evaluation of hemolysis. Hemolysis was less than 30 mg/dl of plasma hemoglobin after 4 h of open chest extracorporeal circulation with 3.0 3.6 L/min of flow rate using the PAD-CP. Other hematologic changes after PAD-CP driving were within normal limits. We conclude that the PAD-CP has proven to have possible clinical applications. PMID- 9212943 TI - Reconsideration of total erythrocyte destruction phenomenon. AB - During a particular long-term in vitro hemolysis test, the plasma free hemoglobin suddenly increased even though the hemolysis level had risen linearly for the previous several hours. This phenomenon was dubbed the total destruction of erythrocytes (TDE) phenomenon, and it was hypothesized that this was the result of the accumulation of sublethal damage to erythrocytes. It was suggested that the TDE might demonstrate the hemolytic characteristics of a pump more sensitively than a conventional hemolysis test. However, the previous report did not consider the effects of temperature or contamination. To study these effects, 3 long-term hemolysis tests were concluded under the following conditions. For Study 1 blood temperature was maintained at 27 degrees C (n = 2); for Study 2, at 37 degrees C (n = 4); and for Study 3, at 37 degrees C with gentamicin (n = 4). The BioMedicus and Nikkiso pumps were used as they were in our previous report. Gas sterilization of all circuits and pumps preceded experimentation. In Studies 1 and 3, hemolysis increased linearly for 29 h. However, in Study 2 a sudden increase of hemolysis occurred for both pumps. Possible causes of this were the dramatic changes in environmental factors such as severe acidosis, high O2 and glucose consumption, and CO2 accumulation. In contrast, neither Study 1 nor Study 3 showed a sudden increase in hemolysis. The plasma free hemoglobin increased linearly in both groups until 29 h of pumping. The environmental changes resulting from contamination were considered to be the cause of the sudden increase in hemolysis. In conclusion, the TDE did not reflect mechanical blood cell damage, but rather different environment situations. Hemolysis increased linearly up to 29 h in either 27 degrees C or germ-free conditions. PMID- 9212944 TI - The method for keeping low perfusion flow weaning from centrifugal pumps: an evaluation of hemolysis in the circulator units. AB - Currently, the clinical application of extracorporeal and assisted circulation using centrifugal pumps is becoming popular. However, a problem in the use of centrifugal pumps is the difficulty in controlling the flow rate when it is a low range. Although we have been controlling the flow rate with a control unit that simply squeezes the blood outlet tube from the outside, hemolysis resulting from turbulent flow in the tube was discovered. Therefore, this control procedure was evaluated experimentally. Two groups were tested as follows: in Group A (n = 10), the tube was squeezed from the outside using the control unit, and in Group B (n = 10), no control unit was used. Blood in each circuit was circulated at 1,500 rpm, and the flow rate was controlled to 250 ml/min using the control unit for Group A. Time-course changes of lactate dehydrogenase (LDH) and free hemoglobin in the blood were measured during 8 h of circulation. There were no significant differences between the 2 groups in the mean values of LDH and free hemoglobin. Therefore, the present low flow rate control provided by the control unit makes it appear to be a simple and useful apparatus for controlling blood circulation using centrifugal pumps. PMID- 9212945 TI - In vitro thrombogenesis study in the Gyro C1E3 for vibration assessment. AB - To clarify the correlation between vibration and thrombus formation in a centrifugal blood pump, a preliminary simulated thrombus study was conducted for possible detection of thrombus formation inside a pump. Additional in vitro thrombogenesis studies were performed to confirm the results of the preliminary study. The primary data acquisition equipment included an accelerometer (Isotron PE accelerometer, Endevco, San Juan Capistrano, CA, U.S.A.), digitizing oscilloscope (TDS 420, Tektronic, Inc., MA, U.S.A.), and pivot bearing centrifugal pumps. The accelerometer was mounted to the top of the pump casing to sense radial and axial accelerations. For the preliminary study, a piece of Silastic was adhered to each of the 3 common areas of thrombus formation inside the pump. The results provided baseline information to speculate on the possibility of detecting thrombus formation by vibration signal changes. For the next studies, fresh bovine blood was harvested under sterile conditions and with strict avoidance of air contact, adding 1.0 U/ml of heparin. The sterilized test circuit consisted of 3/8 inch tubing (Tygon) and a soft reservoir. During the operating time, the activated clotting time (ACT) was maintained between 150 to 300 s using protamin. A restrictor on the outflow tube maintained the flow rates at about 4.5 L/min. The pumps ran continuously for 6 h. Possible blood clot formation inside the pump was monitored by observing the vibration signal from the device for 6 h. These studies revealed that it was possible to distinguish between an impeller that did not form thrombus and ones that formed fibrogenous thrombus using vibration signal assessment. Vibration assessment is worthwhile as a thrombus monitoring tool for a centrifugal blood pump. PMID- 9212946 TI - Quantitative visualization of flow through a centrifugal blood pump: effect of washout holes. AB - To clarify the effect of washout holes on the flow in a centrifugal blood pump to prevent blood stagnation, a quantitative flow visualization technique was applied to compare flows in models with and without washout holes. A scaled-up model of a prototype pump and a high speed video camera were used for the flow visualization, and images were processed by particle tracking velocimetry. Particular attention was paid to the flow through the gaps behind and in front of the impeller. The results showed that in the gap behind the impeller, washout holes caused not only an inward flow, but also an increase in the tangential velocities. In the gap in front of the impeller, washout holes caused an outward flow and a decrease in the tangential velocities. Head flow characteristics were little affected by the washout holes in this initial design for which the flow through the washout holes was set to be approximately 10% of the flow in the external circuit. These results suggest that the flow through washout holes is significant in the prevention of blood stagnation in 2 ways. First, the inward radial velocity behind the impeller and outward velocity in front of the impeller result in fluid exchange, and second, a tangential velocity increase reduces fluid stagnation behind the impeller. PMID- 9212947 TI - Cora rotary pump for implantable left ventricular assist device: biomaterial aspects. AB - Our group is developing a left ventricular assist device based on the principle of the Maillard-Wankel rotative compressor: it is a rotary, not centrifugal, pump that produces a pulsatile flow. Stringent requirements have been defined for construction materials. They must be light, yet sufficiently hard and rigid, and able to be machined with high precision. The friction coefficient must be low and the wear resistance high. The materials must be chemically inert and not deformable. Also, the materials must be biocompatible, and the blood contacting surface must be hemocompatible. We assessed the materials in terms of physiochemistry, mechanics, and tribology to select the best for hemocompatibility (determined by studies of protein adsorption; platelet, leukocyte, and red cell retention; and hemolysis, among other measurements) and biocompatibility (determined by measurement of complement activation and toxicity, among other criteria). Of the materials tested, for short- and middle term assistance, we chose titanium alloy (Ti6Al4V) and alumina ceramic (Al2O3) and for long-term and permanent use, composite materials (TiN coating on graphite). We saw that the polishing process of the substrate must be improved. For the future, the best coating material would be diamond-like carbon (DLC) or crystalline diamond coating. PMID- 9212948 TI - Trial manufacture of eccentric roller type total artificial heart. AB - Working toward a completely implantable total artificial heart, we have designed an eccentric roller type total artificial heart. The actuator of this artificial heart is a drum type eccentric roller that squeezes the blood chambers. The blood chambers are made of silicone rubber and are torus in shape. The shape of the artificial heart is an almost circular cylinder, and its length and diameter are 10 cm and 8 cm, respectively. The 2 main characteristics of this artificial heart are that it discharges blood in a pulsatile mode and that it requires no reversing of the motor. Because we have not completed the artificial heart yet, we have tested the eccentric roller mechanism on the prototype with an overflow type mock circulation with a 100 mm Hg afterload. The prototype worked at the roller speeds of 50, 100, and 150 rpm with flow rates of 1.7, 3.7, and 5.4 L/min, respectively. Next the prototype was connected to a Donovan type mock circulatory system and worked at roller speeds of 88-214 rpm with flow rates of 3.0-8.4 L/min against mean afterloads of 82-120 mm Hg. PMID- 9212949 TI - Magnetically levitated motor for rotary blood pumps. AB - The noncontact rotary pumps under development for use as artificial heart pumps are highly efficient and can prevent thrombus formation. In these pumps magnetic bearings have been widely used to support the rotors to avoid any physical contact. The use of magnetic bearings, however, has led to requirements for the control of a large degree of freedom and for a separate driving motor. This paper introduces 2 types of levitated motors, each of which uses a combination of a rotary motor and a magnetic bearing. These motors are suitable for use in artificial blood pumps because they are small in size and can replace contact components. The radial type levitated motor has the merit of being small in size and capable of controlling the 2 degrees of freedom in the x and y directions. The axial type motor controls only one degree of freedom in the z direction. This paper also introduces the theoretical background of the functions of the motor and magnetic bearing. Experimental results of tests of the proposed motor show a great potential for its application in rotary blood pumps. PMID- 9212950 TI - Development of a novel centrifugal pump: magnetic rotary pump. AB - The rotational axis of the centrifugal pump has some associated problems such as blood destruction and sealing between the axis and pump housing. To improve upon these deficits we have developed a new type of blood pump, the magnetic rotary pump (MRP). The MRP has an original design with no rotational axis and no impellers. We made a prototype MRP and examined its hemodynamics in mock circulation. The prototype MRP flow rate is only 1.0 L/min with an afterload of 30 mm Hg, and we have made some modifications in the size and drive mechanisms from these results. The modified MRP can achieve high flow rates and rotational speeds (6.0 L/min with an afterload of 100 mm Hg, 2,000 rpm) in a mock circuit, and the modified MRP was used for left heart assistance in an acute animal experiment. The MRP could maintain the hemodynamics of an anesthetized adult goat. These results suggest that the MRP needs to be improved in several areas, but the MRP may be useful as a blood pump. PMID- 9212951 TI - Pulmonary arterial impedance analysis by the use of the oscillated assist flow. AB - Pulmonary arterial impedance is an important and interesting characteristic that can be used to evaluate the physiological properties of the pulmonary vessel. However, power spectrum analysis of the pulmonary artery pressure and flow pattern have suggested that peak power in the relatively high frequency range (> 10 Hz) is significantly low; thus, we cannot analyze the vessel properties in the high frequency range. In this study, we used the newly developed vibrating flow pump (VFP), which can generate oscillated blood flow with a relatively high frequency (10-50 Hz) for right heart bypass, to evaluate the pulmonary arterial impedance pattern in the high frequency range. Acute animal experiments of the right heart bypass from the right atrium to the pulmonary artery using 6 healthy adult goats were performed. The flow pattern and pressure of the pulmonary artery, electrocardiograms (ECGs), and arterial and right atrial pressures were continuously monitored during the experiments. Spectral analysis of the hemodynamic parameters using the fast Fourier transform (FFT) method was performed to evaluate the spectral properties. The coherence function, transfer function, and phase patterns were calculated to analyze the impedance pattern in the relatively high frequency area. Previously, various investigators had tried to analyze the impedance patterns of the pulmonary artery; however, they could not analyze the impedance patterns over 10 Hz because the spectral patterns of the pulmonary flow do not have high power at high frequencies. These physiological analyses may be useful in designing the optimal pulmonary circulation. PMID- 9212952 TI - A newly developed silicone-coated membrane oxygenator for long-term cardiopulmonary bypass and cardiac support. AB - The surface of polypropylene hollow fiber was successfully coated with a very thin (0.2 micron) silicone layer. Experimental studies were performed in long term (6 h) normothermic cardiopulmonary bypass (CPB) using 10 goats. A conventional membrane oxygenator (Mera Excelung HPO-15H, MERA, Tokyo, Japan) was used for 5 goats as a control (Group C) and a new silicone-coated membrane oxygenator, which is of the same construction as that of the one used for Group C, for 5 (Group S). The O2 transfer and CO2 removal functions showed the same ranges. In the other parameters, there were no differences between the 2 groups. As for hemolysis, however, the plasma free hemoglobin of Group S was lower than that of Group C. Currently, 3 chronic percutaneous cardiopulmonary support (PCPS) experimental models have been conducted, and there has been no evidence of thromboembolism or deterioration of the oxygenator. In conclusion, this new oxygenator is suitable not only for CPB, but also for long-term cardiac support. PMID- 9212953 TI - Clinical evaluation of the centrifugal pump in open heart surgery: a comparative study of different pumps. AB - The centrifugal pump is now widely used in open heart surgery for its clinical benefits related to the blood elements and the coagulation system. The purpose of this study was to compare the clinical performances of and the outcomes offered by 4 types of centrifugal pumps. For each pump, we investigated the effects on the blood elements, coagulation system, complements, and immunoglobulins during open heart surgery. Four types of centrifugal pumps were used: the HPM-15 (Nikkiso Co.), the Capiox (Terumo Co.), the Lifestream (St. Jude Medical Co.), and the BP-80 (Medtronic, BioMedicus Co.). The platelet count, lactate dehydrogenase (LDH), antithrombin III (AT III), thrombin-antithrombin complex (TAT), complements (C3, C4, and CH50), and immunoglobulins (IgG, IgA, and IgM) were measured before and after cardiopulmonary bypass (CPB). The platelet count was decreased more significantly by the HPM-15 than by any of the other pumps. The other parameters showed no difference among the 4 pumps. In clinical use, each of the 4 types of centrifugal pumps was safe. PMID- 9212954 TI - Cardiopulmonary bypass and cell-saver technique in combined oncologic and cardiovascular surgery. AB - The purpose of this study was to work out an adequate operative technique for patients with malignant tumors who also need open heart surgery or procedures on major blood vessels. We had 8 such patients. In 6 of them, a tumor (3 cases hypernephroid cancer and 3 cases retroperitoneal sarcoma) had grown through the inferior vena cava (IVC) up to the right atrium. Two patients had lung cancer together with severe coronary artery disease. All of these patients were operated on using a heart-lung machine (HLM) and cell saver (CS). In 6 patients the intravascular portion of the tumor was extracted as much as possible through a right atrium approach (in 3 cases a nephrectomy was performed). Two patients had a one-stage coronary artery bypass graft (CABG) and a lobectomy. All of the patients had uneventful postoperative periods and were alive when checked on 1 year after the procedures. During cytological investigation after each operation, tumor cells were found only on the internal surface of the HLM arterial filters with 20 microns holes. We suggest that special cardiovascular devices such as the HLM and CS might be used in borderline situations in oncology without increasing the risk of hematogenous tumor dissemination. PMID- 9212956 TI - A clinical study for the durability of oxygenators on cardiopulmonary support. AB - Cardiopulmonary support (CPS) requires durability of the oxygenator. The life span of the oxygenator is affected by various clinical factors, including patient condition, perfusion condition, and equipment usage. Predictors for the durability of oxygenators were evaluated clinically in this study. Thirty-two patients, who had undergone CPS during the last 3 years in our institute were assigned to this study. Fifty oxygenators had been used (Capiox SX in 19, CB Maxima in 23, and AL-6000 in 8). Significant predictors for the durability of oxygenators were evaluated by nonparametric survival analysis and proportional hazards regression analysis. Univariate regression analysis revealed 6 significant predictors for the life span of oxygenators. These were the oxygenator type, type of centrifugal pump, acidosis with blood pH less than 7.35, base excess less than -5, blood glutamic-oxaloacetic transaminase (GOT) levels greater than 1,000 IU, and blood lactate dehydrogenase (LDH) levels greater than 3,000 IU. After multivariate analysis, there remained only 2 significant predictors. An oxygenator used with a noncoated CPS system (Capiox SX with Capiox EBS) proved to have a significantly shorter life span than one used with a heparin-coated system (CB Maxima or AL-6000 with CB BP-80) (hazards ratio, 3.588, p = 0.0065). Patient conditions, which revealed acidosis with less than -5 of base excess, significantly shortened the life of the oxygenator (hazards ratio, 3.595, p = 0.0188). PMID- 9212955 TI - Various problems during long-term percutaneous cardiopulmonary support. AB - A 54-year-old man with a left ventricular free wall rupture following acute anterior myocardial infarction underwent a repair surgery with percutaneous cardiopulmonary support (PCPS). During surgery and postoperatively, PCPS provided sufficient support flow. The patient was successfully weaned from PCPS on the 15th postoperative day and discharged subsequently. In the management of cardiac rupture patients, PCPS has the merit of preventing rupture progression and the advantage of recovery of pulmonary function. However, there are several problems to solve. The support effectiveness and recovery of the patient's heart should be carefully evaluated. Effective left heart decompression also needs to be established. Heparin-coated circuits still need proper anticoagulation treatment to prevent thrombus formation especially while support flow is low. A circuit construction that allows easier maintenance and safer exchange of oxygenators and pump heads is suggested. Ischemia of the cannulated leg should be prevented by femoral artery perfusion. PMID- 9212957 TI - Experimental studies on three types of heparin-coated cardiopulmonary bypass circuits. AB - In cardiovascular operations, we have usually used heparin-coated cardiopulmonary bypass circuits with low systemic heparinization. Three types of heparin-coated cardiopulmonary bypass circuits are available in Japan: 2 of the 3 have covalent heparin bonding, and the other has ionic heparin bonding. We studied these circuits in ex vivo experiments to explore which were the best in terms of biocompatibility. In this study we compared the Carmeda system (Medtronic) and the Capiox system (Terumo) with covalent heparin bonding, and the Duraflo-II (Baxter) with ionic heparin bonding, evaluating them in ex vivo experiments. They were primed with fresh human blood, and we studied and compared the platelet counts, fibrinogen, D-dimmer, beta-thioguanine (TG), thrombin-antithrombin complex (TAT), and C3a and C4a of each of them. Additional research will be presented in the future. PMID- 9212958 TI - Comparison of the Gyro C1E3 and BioMedicus centrifugal pump performances during cardiopulmonary bypass. AB - The compact eccentric inlet port (C1E3) centrifugal blood pump was developed as a cardiopulmonary bypass (CPB) pump. The C1E3 pump incorporated a sealless design with a blood stagnation free structure. The pump impeller was magnetically coupled to the driver magnet in a sealless manner. To develop an atraumatic and antithrombogenic centrifugal pump without a shaft seal junction, a double pivot bearing system was introduced. Recently, a mass production model of the C1E3 was fabricated and evaluated. The ratio of the normalized index of hemolysis (NIH) of the C1E3 was 0.007 g/ 100 L, in comparison to the NIH of the BP-80, 0.018 g/ 100 L, each in a CPB condition of 5 L/min against 325 mm Hg. Both pumps were compared in identical in vitro circuits. To further evaluate the pumps during cardiopulmonary bypass for reliability and function, 6 h of CPB was performed on each of 8 bovines using either the C1E3 or BP-80 centrifugal pump. The BP-80 and C1E3 provided pump flows of 50-60 ml/kg/min without incident. The hemodynamics were stable, and the hematology and biochemistry data were within normal ranges. There were no statistically significant differences between the 2 groups. Concerning the plasma free hemoglobin values, a mass production model of the C1E3 pump had the same hemolysis levels as the BP-80. Our preliminary studies reveal that the C1E3 pump is reliable. Also, the C1E3 will satisfy clinical requirements as a cardiopulmonary bypass pump. PMID- 9212959 TI - Effective cross-circulation technique of venoarterial bypass for differential hypoxia condition. AB - We examined a new technique of cross-circulation (CC) venoarterial bypass (VAB) with femoral arterial perfusion and superior vena cava drainage through a long femoral venous cannula. Six adult mongrel dogs weighing 15 to 20 kg underwent the CC-VAB with oxygenation after introduction of respiratory failure (RF). The flow of the CC-VAB was maintained at half the level of the control cardiac output, and the hemodynamic parameters were monitored. To evaluate hypoxia in the upper body, the arterial partial pressure of oxygen (PaO2 [mm Hg]) in the carotid artery and the venous saturation of oxygen (SvO2 [%]) in the pulmonary artery were measured during control, RF, standard VAB, and CC-VAB conditions. The PaO2 decreased significantly after the introduction of RF (41.7 +/- 12.4), and it returned to normal levels only after CC-VAB (151.2 +/- 24.5, p < 0.05). The SvO2 during CC VAB (98.6 +/- 2.1) was significantly higher than that during VAB without CC (53.5 +/- 3.4, p < 0.05). These results suggest that this cross-circulation technique could be applied to patients with differential hypoxia during femoral VAB with oxygenation or percutaneous cardiopulmonary support (PCPS). PMID- 9212960 TI - Significant left ventricular unloading with transaortic catheter venting during venoarterial bypass. AB - Insufficient unloading of the left ventricle with blood stagnation is a main cause of unsuccessful left ventricular (LV) recovery during percutaneous cardiopulmonary support (PCPS). The purpose of this investigation was to evaluate the effectiveness of transaortic catheter venting (TACV) for LV unloading. Six adult mongrel dogs (mean weight 16.3 kg, range 14-20 kg) underwent venoarterial bypass (VAB) with TACV. Bypass flow ranged from 0.8-1.2 L/min, and TACV flow ranged from 160-240 ml/min. In addition to monitoring the standard hemodynamic parameters, the slope of the LV end-systolic pressure-volume relation (Emax) during transient occlusion of the inferior vena cava, the slope of the LV end systolic pressure-stroke-volume relation (Ea), the stroke work (SW), the LV pressure-volume area (PVA), and the slope of the SW end-diastolic volume relation, the preload recruitable stroke work (PRSW) were assessed by means of a microtip manometer and a conductance catheter. The LV contractility (Emax) and aortic elastance (Ea) were equivalent in the 2 groups with or without TACV (7.7 +/- 1.1 versus 8.4 +/- 1.5 mm Hg/ml and 8.2 +/- 1.4 versus 7.6 +/- 1.3 mm Hg/ml). Comparing the measurements for the baseline to those for VAB with TACV, the SW was significantly reduced, and the PVA/SW was increased by TACV (1,685 +/- 309 versus 867 +/- 188 x 10(-4) J, p < 0.05 and 1.32 +/- 0.03 versus 1.58 +/- 0.11, p < 0.05, respectively). Furthermore, the PRSW was gradually decreased from the baseline value to the value resulting from VAB with TACV (75 +/- 8 versus 44 +/- 3 x 10(-4) J/ml, p < 0.01). In comparison, the percent reduction of SW between VAB and VAB with TACV tended to be increased by TACV (23.2 +/- 7.2% versus 46.9 +/- 7.7%, p = 0.05). These results suggest that TACV might reduce LV work (SW and PRSW) and might increase the LV energetic charge. In conclusion, TACV would be an adjunctive technique to VAB or PCPS for patients with LV failure. PMID- 9212961 TI - Single needle venovenous extracorporeal membrane oxygenation using a nonocclusive roller pump for rescue in infants and children. AB - In 1993, J.Y. Chevalier described a single needle venovenous extracorporeal membrane oxygenation (ECMO) system using a nonocclusive roller pump and alternating clamps for pulmonary support in neonates. We modified this system to use it in older children as well and for additional indications. Introducing a double raceway and 2 different sizes of tubing sets and performing percutaneous approach, we treated 21 children (age 1 day to 49 months) using this system. Indications for treatment were hypoxia and hypoxic induced myocardial dysfunction resulting from pulmonary failure, sepsis, and congenital defects. Of the children treated for neonatal indications, 7/9 survived. For 2 children ECMO was terminated because of intraventricular hemorrhage (IVH). In the pediatric group 5/7 of the children could be weaned from ECMO, and 2 children died after more than 30 days on ECMO. Two of the children who had been almost completely weaned died later because of therapy withdrawal following a brain death diagnosis. In the cardiac group, 3/5 of the children survived. We conclude that the described system is an effective venovenous ECMO system that reduces invasivity and expenditure. PMID- 9212962 TI - Recent experience of integrated myocardial management: the newest strategy for myocardial protection. AB - From April 1994 until June 1996, we exclusively utilized the integrated myocardial management (IMM) proposed by Buckberg et al. at UCLA. Two hundred sixty-two consecutive patients undergoing open heart surgery at our hospital were divided into 2 groups, the non-IMM (n = 49, from July 1993 until March 1994) and the IMM (n = 213, from April 1994 until June 1996) groups. Although many older and more severely ill patients were treated with IMM, acceptable clinical outcomes with comparable safety and efficiency were obtained. Shorter durations of total cardiopulmonary bypass (CPB) and aortic cross-clamping (AXC) were needed in the IMM group despite there being many more procedures undertaken during a single cross-clamp period. PMID- 9212963 TI - Clinical study of totally roller pumpless cardiopulmonary bypass system. AB - We have developed a low negative pressure vacuum suction system in which cardiotomy suction is performed by the negative pressure of the venous reservoir controlled by a vacuum controller. We have employed this vacuum suction system with a centrifugal pump as a totally roller pumpless cardiopulmonary bypass (CPB) system. In this study, the clinical availability and hemocompatibility of our totally roller pumpless CPB system were evaluated by a randomized prospective study. Thirty patients undergoing aortocoronary bypass grafting were assigned to the study. Data from seventeen patients treated with a totally roller pumpless CPB system were compared with data from 13 treated with a conventional roller pump CPB system. Totally roller pumpless CPB reduces hemolysis, showing lower plasma free hemoglobin levels (81.8 +/- 25.0 versus 42.0 +/- 16.3 at 30 min after CPB initiation, p < 0.05), higher plasma haptoglobin levels (37.8 +/- 36.6 versus 77.2 +/- 31.3 at 120 min after CPB, p < 0.05), and lower blood lactate dehydrogenase (LDH) levels (1391 +/- 497 versus 972 +/- 187, p < 0.01) than those of CPB with a roller pump suction with no significant difference between platelet counts. Arterial blood oxygen tension after using a totally roller pumpless CPB system was slightly better than that with a roller pump (396 +/- 48 versus 437 +/ 43, p = 0.069); however, there was no significant difference in intubation times between groups. A totally roller pumpless CPB system provides sufficient biocompatibility for the blood to reduce hemolysis significantly and simplifies and miniaturizes the entire CPB system to achieve good visuality and handling for control as well. PMID- 9212964 TI - Flow measurement at the pump head of centrifugal pumps: comparison of ultrasonic transit time and ultrasonic Doppler systems. AB - Determination of blood flow is essential for monitoring rotary blood pumps. However, accurate measurement directly adjacent to the pump housing is difficult because of the highly irregular flow profiles near the fast spinning rotor. Therefore, a specially adapted flow probe based on the ultrasound transit time (USTT) principle was designed to evaluate the flow in centrifugal blood pumps. The probe can be directly mounted at the housing and creates 2 crossed measuring ultrasound beams. The mean value, Qm, of the 2 output signals corresponds to the blood flow and the difference, Qd, correlates to the vorticity of the flow profile in the pump outflow tract. In vitro measurements obtained an accuracy for mean flow values of better than +/-0.6 L/min in extreme working points and for vorticity values even as high as Qd = 3.5 L/min. Because of vorticity, however, the output signal contained considerable noise, and that required the application of a 10 Hz filter. Positioning of the ultrasound (US) beams parallel to the axial direction of the pump was superior to radial positioning. Additional measurement of the flow profile demonstrated that a large vorticity occurred (up to Qd equal to 3.5 L/min), and this vorticity was highly dependent upon the afterload of the pump. In vivo experiments demonstrated the reliability of the method. We concluded that USTT flow measurement can determine blood flow immediately adjacent to the pump housing with sufficient accuracy, and these measurements are superior to those from US-Doppler systems (which cannot handle the vorticity accurately enough) and electromagnetic devices (which lack zero stability). PMID- 9212965 TI - Clinical evaluation of pulsatile flow mode of Terumo Capiox centrifugal pump. AB - The Terumo Capiox centrifugal pump system possesses an automatic priming function in which the motor repeatedly stops and runs intermittently to eliminate air bubbles in the circuit through the micropores of the hollow-fiber membrane oxygenator. By modifying this mechanism, we have developed a pulsatile flow mode. In this mode, maximum and minimum pump rotational speeds can be independently set every 20 rpm in the range of 0 to 3,000 rpm. The duration of the pump run at maximum and minimum speeds can also be independently set every 0.1 s in the range of 0.2 to 15 s. In a clinical trial, after obtaining the desired flow rate, 2.4 L/min/m2 in nonpulsatile flow mode, a pulsatile flow mode of 60 cycles/min (with 1 cycle being maximum speed for 0.4 s and minimum speed for 0.6 s) was obtained by adding and subtracting 500 rpm to and from the rotational speed in nonpulsatile flow mode. Pulse pressures in the femoral artery and in the circuit just proximal to the perfusion cannula (6.5 mm Sarns high flow cannula with metal tip) were measured in 5 patients who underwent pulsatile cardiopulmonary bypass (CPB) for a coronary artery bypass graft (CABG), and compared to pulse pressures obtained by intraaortic balloon pumping (IABP) in 3 patients and by the pulsatile mode of the 3M Delphin pump in 3 patients. The platelet count, free hemoglobin, and beta-thromboglobulin (beta-TG) were measured and compared with measurements from another 5 patients who underwent nonpulsatile CPB. Although the pulse pressure measured in the circuit was 180 mm Hg on average, the pressure in the femoral artery was only 15 to 40 mm Hg with a mean of 20 mm Hg. In the same patients, 60 to 80 mm Hg pulse pressure was obtained with IABP. The pulse pressure obtained with the Delphin pump was not more than that obtained with the Terumo pump. There were no significant differences in percents of preoperative levels of platelet counts (pulsatile, 87.6 +/- 15.8% and nonpulsatile, 72.4 +/- 40.6%), free hemoglobin (pulsatile, 18 +/- 8 mg/dl and nonpulsatile, 25 = 7 mg/dl), and beta-TG (pulsatile 298 +/- 28 ng/ml and nonpulsatile, 312 +/- 143 ng/ml). In conclusion, although the pulsatile mode of the Terumo centrifugal pump did not exhibit any adverse effects hematologically, the pulse pressure obtained was unsatisfactorily small, mainly because of dumping caused by the perfusion cannula. PMID- 9212966 TI - Our distal aortic perfusion system in descending thoracic and thoracoabdominal aortic aneurysm repairs. AB - We have used heparin-bonded partial cardiopulmonary bypass to support distal aortic circulation during aortic cross-clamping. However, there were no cardiotomy reservoirs with fully reliable thromboresistance. To resolve this problem, a short-acting anticoagulant (nafamostat mesilate) was added into a cardiotomy reservoir. The present study was designed to evaluate the efficacy of our distal perfusion system. From May 1995 through the end of May 1996, 27 patients underwent descending thoracic and thoracoabdominal aortic aneurysm repairs with this adjunct, 4 being excluded from the experiment. Twenty patients who had undergone conventional partial cardiopulmonary bypass were defined as the control group. There were no significant differences between the 2 groups in the morbidity, mortality, gas transfer, or transfusion requirements despite the fact that more complicated surgical procedures (shown by a two-fold increase in the prevalence of reoperation) were required in the group that had received the current distal perfusion adjunct the heparin-bonded group. In conclusion, our perfusion system is very effective for descending thoracic and thoracoabdominal aortic aneurysm repairs. PMID- 9212967 TI - Prevention of remote organ injury in cardiopulmonary bypass: the impact of flow generation technique. AB - The purpose of this study was to investigate the effects of 3 different types of flow generation for cardiopulmonary bypass on gastrointestinal permeability and on neutrophil expression of CD11b, a surface marker of neutrophil activation. Fourteen patients undergoing elective coronary revascularization were selected randomly to receive 1 of the 3 flow generation techniques (roller, pulsatile, or centrifugal). Intestinal permeability was assessed by the fraction of an oral dose of 51chromium-ethylenediaminetetraacetate (51Cr-EDTA) recovered in the urine over 24 h. Neutrophil activation was determined by expression of CD11b markers at 6 time points. Overall, the 14 patients showed significant increases in intestinal permeability. It was not possible to demonstrate statistically significant differences among the flow generation groups; however, when compared to both roller pump groups, the centrifugal pump group showed a 3.2% reduction in intestinal permeability. There was no change in the expression of CD11b receptors throughout the time points, nor was there a relationship of CD11b markers to the flow generation technique. PMID- 9212969 TI - Biocompatibility of heparin-coated circuits in pediatric cardiopulmonary bypass. AB - In this study, we evaluated the biocompatibility of heparin-coated circuits in pediatric cardiopulmonary bypass (CPB). Eight patients were divided into 2 groups: the control group (Group C) and heparin-coated group (Group H). In Group H, CPB circuits, including the arterial pump, oxygenator, and cannulas were heparin-coated. Before, during, and after CPB, blood samples were obtained to assess the platelet counts (Plat), alpha 2-plasmin plasminogen inhibitor complex (PIC), thrombin-antithrombin III complex (TAT), C3 activation products (C3a), interleukin (IL)-6, IL-8, and polymorphonuclear neutrophil leukocyte (PMN) elastase. There was no significant difference in Plat, PIC, or TAT between groups. Group H showed significantly low levels of C3a (during and after CPB), PMN elastase (during CPB), and IL-6 (after CPB). These data demonstrated that in pediatric CPB, heparin-coated CPB circuits reduced the activation of complements and the production of PMN elastase and IL-6, suggesting the superior biocompatibility of the heparin-coated circuits. PMID- 9212968 TI - Renal circulation and cellular metabolism during left ventricular assisted circulation: comparison study of pulsatile and nonpulsatile assists. AB - We examined left ventricular assist during 6 h for an acute myocardial infarction model in pigs. The outflow cannula was placed in the ascending aorta and an inflow cannula in the left atrium. A pump (Pulsatile group: Zeon Medical and Nonpulsatile group: Nikkiso HPM-15) was connected to each cannula. Items measured were the regional blood flow of the cortex and the medulla in the kidney, renal arterial flow, arterial blood ketone body ratio (AKBR), lactate/pyruvic acid, BUN, creatinine and beta 2-microglobulin. After experimental study, the kidneys were removed, and a pathological study was performed. In the pulsatile assisted group, renal cortical blood flow increased but medulla blood flow decreased. On the other hand, in the nonpulsatile assisted group, both regional blood flows decreased. That means that in the pulsatile assisted group intrarenal redistribution improved, rather than in the nonpulsatile assisted group. The results of our study indicated that pulsatile assist produced superior circulation in the kidney, and the microcirculation on the cell level was superior as well in early treatment of acute left heart failure. PMID- 9212970 TI - A new blood pump for cardiopulmonary bypass: the HiFlow centrifugal pump. AB - Centrifugal blood pumps are considered to be generally superior to the traditionally used roller pumps in cardiopulmonary bypass. In our institute a new lightweight centrifugal sealless blood pump with a unique spherical thrust bearing and with a magnetic coupling was developed, the HiFlow. The small design makes the pump suitable for applications in complex devices or close to a patient. Hemolysis tests were carried out in which the BioMedicus pump BP-80 and a roller pump were used as reference. The centrifugal pump HiFlow showed the least blood trauma within the group of investigated pumps. In summary, the HiFlow pump concept with its low priming volume and limited contact surfaces shows great potential for clinical applications in cardiopulmonary bypass. Also, the possibility of using the pump as a short-term assist device with an option of a pulsatile driving mode was demonstrated. PMID- 9212971 TI - Use of percutaneous cardiopulmonary support system with rotary blood pump in graft replacement of the descending thoracic and thoracoabdominal aorta. AB - Graft replacement of the descending thoracic or thoracoabdominal aorta was successfully performed in 3 patients using percutaneous cardiopulmonary bypass. Femoral inflow and outflow cannulas were inserted percutaneously after induction of anesthesia with the patient in supine position, and low flow normothermic bypass was established before thoracotomy. Next the patient was placed in a right lateral position to create an operating field. With this body position and even an almost prone position, which was sometimes necessary for easy dissection of adhesion of lung to the aneurysmal wall, the bypass flow was easily maintained adequately. The bypass circuit was coated with heparin, and the activated clotting time (ACT) was controlled to be between 150 and 200 s during the entire operating period. Percutaneous insertion of the cannulas avoided local bleeding in the groin, and the low ACT made control of hemorrhage in the operating field easy. For descending aortic surgery, heparin-coated percutaneous cardiopulmonary bypass proved to be a useful adjunctive measure. PMID- 9212972 TI - Study of regional cerebral oxygen saturation during percutaneous cardiopulmonary support. AB - The purpose of this study was to evaluate the change of regional cerebral oxygen saturation (rSO2) during percutaneous cardiopulmonary support (PCPS) in patients with cardiogenic shock. Fifteen patients with cardiogenic shock were evaluated during PCPS by continuous monitoring of rSO2, systemic venous oxygen saturation (Svo2), and hemodynamics. The brain damage of these patients was also evaluated during and after PCPS. There were 10 males and 5 females. Their ages ranged from 57 to 79 years old (average: 60.0 +/- 14). Two patients were unconscious before PCPS, and 11 received intraaortic balloon pumping (IABP) before PCPS. The change of rSO2 was significantly correlated with the change of Svo2. The average of rSO2 was 64 +/- 3% at the stable hemodynamic condition. The rSO2 with pulsatile PCPS was higher than that with nonpulsatile PCPS. There was no correlation between brain damage and rSO2. The patients with low rSO2 (< 50%) that resulted in poor LV function could not be weaned from PCPS. In conclusion, the continuous monitoring of rSO2 during PCPS could be a useful tool. PMID- 9212973 TI - Application of a licorice root specific protein to a general method for the assay of licorice root components in traditional Chinese medicines. AB - A new fuzzy immunoassay method generally applied to ten Glycyrrhizae Radix (GR) preparations of four different botanical origins was studied. Four kinds of antisera were elicited in rabbits immunized with GRs of different botanical origins. The presence of the characteristic GR protein (GRP) was shown using Western blot analyses and selected antibody enzyme immunoassay (SAEIA) methods. A GRP was isolated from one of the GR specimens which was selected using SAEIA methods. The isolated GRP was heated to reduce its binding activity to an anti-GR serum. A new fuzzy SAEIA method generally applicable for assay of the extract of the ten GR specimens was developed using heat-treated GRP as the solid-phase antigen. The fuzzy SAEIA method was successfully applied for the detection and quantitative analysis of the GR component contained in traditional Chinese medicines. PMID- 9212974 TI - In vitro glucuronidation of 25-hydroxyvitamin D3 and its pro-form. AB - In vitro glucuronidation of 25-hydroxyvitamin D3 and its pro-form has been investigated by means of HPLC with UV detection. Although both substrates gave 3- and 25-glucuronides in the presence of the rat liver microsomal fraction and uridine-5'-diphosphoglucuronic acid, 25-hydroxyvitamin D3 and its pro-form yielded 3- and 25-glucuronide as the main product, respectively. The latter glucuronide is the one in which the tert-hydroxy group is conjugated. Each glucuromide was identified by its chromatographic behavior in comparison with an authentic sample and data obtained from liquid chromatography/mass spectrometry (LC/MS) using atmospheric pressure chemical ionization. PMID- 9212975 TI - Does small-dose gamma-ray radiation induce endogenous antioxidant potential in vivo? AB - The induction of in vivo antioxidant potential following small doses of gamma-ray irradiation was investigated in C57BL/6 mice. The antioxidant capacity of various organs was assessed in terms of the scavenging activity of cytosol fractions against 1,1-diphenyl-2-picrylhydrazyl (DPPH), a chemically stable radical. Significant elevations in the scavenging activity were recognized in several organs, including the liver, pancreas and brain, soon after post-irradiation with 50 cGy of gamma-ray. These increases persisted for 24 h. gamma-Radiation of the liver at 25-50 cGy elevated its cytosolic antioxidant capacity, but this was lowered at 200 cGy. In order to assess which antioxidants underlie this phenomenon, the content of a reduced form of glutathione (GSH) in liver was assayed as a function of time after gamma-irradiation at a dose of 50 cGy. The GSH content was significantly increased from 6 h after irradiation, and this change was consistent with that of the total radical scavenging potency of the liver against DPPH. Further, the elevation of GSH content was accompanied by elevated GSSG reductase activity induced by gamma-irradiation. PMID- 9212976 TI - Effects of hyperosmotic NaCl and glycerol stress on stress response of human HeLa cells. AB - The maintenance of intracellular osmotic pressure is of fundamental importance for cell survival. Since osmoregulatory processes are important to all living organisms, large fluctuations in environmental osmolarity may elicit or modulate stress responses. We examined whether hyperosmotic stress induced or modulated stress responses in human HeLa cells. When HeLa cells were incubated in medium supplemented with 50-150 mM NaCl or 0.2-1.0 M glycerol for 3-27 h, the stress response, analyzed at the levels of HSP70 synthesis, HSP70 mRNA accumulation, and heat shock transcription factor (HSF) activation, was not induced by either hyperosmotic stress. In hyperosmotic 150 mM NaCl or 1.0 M glycerol medium, the stress response to heat shock was inhibited at the levels of HSF activation, HSP70 mRNA accumulation, and HSP70 synthesis. In vitro activation of HSF showed that inhibition of this activation by hyperosmotic NaCl or glycerol stress was not irreversible. Furthermore, addition of the physiological osmolyte betaine to medium reversed the inhibition of heat-induced HSP70 synthesis under hyperosmotic NaCl stress but not under hyperosmotic glycerol stress. The effect of betaine against hyperosmotic NaCl stress was observed mainly at the translation level, and betaine seemed to enable cells to translate HSP70 mRNA specifically under heat shock conditions by restoring protein synthetic ability. PMID- 9212977 TI - Effect of transcriptional regulatory sequences on autonomous replication of plasmids in transient mammalian systems. AB - Transcriptional regulatory sequences often influence the efficiency of DNA replication, directly or indirectly, in bacteria, yeast, and animal virus systems. We have tested several transcriptional regulatory sequences for affecting DNA replication, based on pUC vector, in transient systems. Autonomous replication of transfected plasmids was assayed by PCR amplification of the fragments derived from the plasmids, which had replicated in mammalian cells. By this highly efficient method of detecting replicated molecules, pUC19, but not pUC18, showed a week replication activity in transfected cells. Nucleotide sequences around the HindIII site in pUC19 were required for replication. Monomers or dimers of the octamer transcription motif of the mouse immunoglobulin heavy chain gene, inserted in multicloning sites of pUC19, could stimulate replication, while the 4- or 6-mers did not, in contrast to the results on its transcription activity. Other transcriptional elements including AP1, HSE, and E2F also stimulated replication, but neither CRE nor Sp1 binding motif did. These results suggest that at least some of the transcriptional regulatory sequences function as-modulators of DNA replication as well as of transcription. PMID- 9212978 TI - Na+(Li+)/H+ antiporter in Pseudomonas aeruginosa and effect of Li+ on cell growth. AB - Everted membrane vesicles were prepared from cells of Pseudomonas aeruginosa, and cation/H+ antiport was measured. We observed activities of Na+/H+ antiport, Li+/H+ antiport and K+/H+ antiport. Judging from the competition pattern, it seems that there are at least two types of antiporter, a Na+(Li+)/H+ antiporter and a K+/H+ antiporter. Na+ was a good substrate for the Na+(Li+)/H+ system, whereas Li+ was a poor substrate. Although the K(m) value for Na+ (or Li+) was similar to those in Escherichia coli Na+/H+ antiporters, the Vmax value for Na+ (or Li+) was much smaller in the P. aeruginosa antiporter than in the E. coli antiporters. Growth of P. aeruginosa was strongly inhibited by 0.4 M LiCl, but not by NaCl or KCl. PMID- 9212979 TI - The potent inhibition of vapiprost, a novel thromboxane A2 receptor antagonist, on the secondary aggregation and ATP release of human platelets. AB - The inhibitory effects of vapiprost hydrochloride (vapiprost), a novel thromboxane A2 receptor antagonist, on platelet aggregation and ATP release were studied using platelet rich plasma (PRP) of humans, guinea pigs, rabbits and rats. In in vitro experiments with human platelet, vapiprost inhibited the aggregation and ATP release stimulated with U-46619, collagen or arachidonic acid (AA) at an IC50 of less than 2.1 x 10(-8) M. Vapiprost did not inhibit the primary aggregation or ATP release of human platelets stimulated with adenosine 5'-diphosphate (ADP), epinephrine (Epi) or platelet activating factor (PAF), but inhibited the secondary aggregation stimulated with those agonists at an IC50 of less than 1.3 x 10(-7) M. The sensitivity of platelets in various species of animals to vapiprost was in the following order: human > or = guinea pigs > rats > rabbits. In ex vivo experiments with guinea pigs which received a single oral dose of vapiprost, the agent demonstrated strong inhibition of ATP release from platelets stimulated with U-46619, collagen or AA at an ID50 of less than 25.8 micrograms/kg. These inhibitory effects were observed within 30 min and sustained for 24 h at a single dosage of 5 mg/kg of vapiprost. In AA-induced pulmonary infarction models of mice, the sudden death rates decreased significantly with the oral administration of 10 mg/kg or more of vapiprost. These results indicate that vapiprost effectively inhibits the secondary aggregation and ATP release of human platelets stimulated with various agonists, and that guinea pig and human platelets are similar in response to vapiprost. Furthermore, it was demonstrated in ex vivo experiments with guinea pigs that the inhibitory action of vapiprost appears rapidly and lasts for long periods. PMID- 9212980 TI - Ascorbic acid 2-O-alpha-glucoside-induced redox modulation in human keratinocyte cell line, SCC: mechanisms of photoprotective effect against ultraviolet light B. AB - We previously reported that the topical application of ascorbic acid 2-O-alpha glucoside (AA-2G) suppressed the cutaneous inflammation by ultraviolet irradiation in human and guinea pigs (Miyai et al., Nishinihon J. Dermatol., 58, 439-443 (1996)). In this paper, the effect of AA-2G on the lethal damage induced by ultraviolet B (UVB) was studied using a human keratinocyte cell line, SCC, established from squamous cell carcinoma. The photoprotective effect of AA-2G on cytotoxicity of UVB in SCC cells was dose dependent (0.125-1 mM) and more effective than that of ascorbic acid (AsA) at 1 mM. This protection was completely abolished in the presence of an alpha-glucosidase inhibitor, castanospermine, indicating that release of AsA from this derivative was essential for reduction of the actinic injury. AA-2G significantly suppressed cytotoxicities of hydrogen peroxide and superoxide anion produced by xanthine and xanthine oxidase. AA-2G exhibited a preventive effect against the cytotoxicity produced by tert-butylhydroperoxide, an inducer of lipid peroxidation, in the presence of alpha-tocopherol, but not in the absence of alpha-tocopherol. Cytotoxicity of UVB was also effectively reduced by the combination of AA-2G and alpha-tocopherol. In addition, AA-2G reduced UVB-promoted formation of lipid peroxide and accumulation of lipofuscin, which is known to be a complex of cellular proteins and metabolites of lipid peroxide. These data suggest that AA 2G prevents the acute inflammation induced by UVB irradiation partly through scavenging reactive oxygen species and potentiating the antioxidative activity of alpha-tocopherol. PMID- 9212981 TI - Anti Candida activity of induced transferrin in mice immunized with inactivated Candida albicans. AB - Mice immunized with formalin-killed Candida albicans were resistant to challenge by a lethal amount of viable C. albicans. The growth-inhibitory activity to C. albicans was detected in sera from the immunized mice, and was inhibited by the addition of anti-transferrin antibody or ferric sulfate. Both the amount of transferrin and the unsaturated iron-binding capacity (UIBC) in the serum were significantly increased, indicating that apo-transferrin increased in the immunized mice. Moreover, the intraperitoneal administration of apo-transferrin enhanced the protection from the Candida infection in vivo. PMID- 9212982 TI - Involvement of calciseptine-sensitive calcium channels in the evoked acetylcholine release from electric organ synaptosomes. AB - The high K(+)-evoked release of acetylcholine (ACh) from electric organ synaptosomes isolated from the Japanese marine ray, Narke japonica, was strongly inhibited by dihydropyridines at micromolar concentrations. However, this inhibition seems to be a non-specific effect since the agonist (-)-Bay K 8644 also had inhibitory effects. Calciseptine, a peptide toxin specific for L-type Ca channels, inhibited to a lesser extent the evoked acetylcholine release: the maximum inhibition was about 20%. This finding is in accord with our data (Tokumaru et al., J. Neurochem., 65, 831(1995)) regarding inhibition by a monoclonal antibody against the alpha 2 delta-subunit of the L-type Ca channel and provides evidence for the involvement of an L-type-like Ca channel in ACh release in addition to omega-conotoxin GVIA-sensitive N-type and omega-agatoxin IVA-sensitive P-type Ca channels. PMID- 9212983 TI - Dual stimulatory and inhibitory effects of fluorine-substitution on mutagenicity: an extension of the enamine epoxide theory for activation of the quinoline nucleus. AB - Nineteen mono- and di-fluorinated derivatives of quinoline, 1,7-phenanthroline, 1,10-phenanthroline, benzo[h]quinoline, and benzo[f]quinoline were subjected to analysis of their structure-mutagenicity relationships. For this purpose, six new fluorinated derivatives were synthesized. The results support that the enamine epoxide structure of the pyridine moiety, as well as the bay-region epoxide structure, is responsible for mutagenicity. Formation of K-region epoxides might involve a detoxification process rather than mutagenic activation. PMID- 9212984 TI - Electrophoretic patterns of urinary proteins of diabetics in the pre, early and overt nephropathy stages. AB - Urinary proteins of patients with diabetes mellitus (DM) were analyzed using cellulose acetate membrane electrophoresis, to determine the clinical usefulness of fraction patterns of the proteins in detecting the group at high risk for diabetic nephropathy. We divided the protein patterns into 5 groups. Four groups (I, II, III, IV) were found in the healthy group and a newly classified group was termed group 0 and was characterized by a prominent albumin peak with a negligible or small globulin peak. The incidence of groups 0, I, II, III, and IV, was 36.6%, 13.3%, 18.7%, 10.7% and 22.7%, respectively. This distribution was clearly different from that of healthy subjects and the most characteristic feature of diabetics was that group 0 accounted for 36.6% of the total cases. Characteristic features of each group were examined from the aspect of laboratory and clinical findings. Urinary protein patterns were concluded to be useful not only to predict the high risk group for diabetic nephropathy in the preclinical stage but also to discriminate nephropathic types of glomerular or tubular origin. It is useful for clinicians to know the risk stage and prognosis for diabetic nephropathy. PMID- 9212985 TI - Plasma triglyceride-decreasing components of pine needles. AB - An ultrafiltrate (M.W. < 10000) of 0.1 M acetic acid extract from Japanese red pine needles was treated with Sep-Pak Vac C18, and an absorbable fraction (pine aqueous components fraction, PAC) was obtained. Since the intraperitoneal administration of PAC to rats decreased plasma triglyceride in a screening test for bioactivity, we tried to further isolate the active substances with respect to this triglyceride-decreasing action. Active substances were precipitated by acid, and we then divided them into fractions using reversed phase HPLC. The active fractions were hydrolyzed, then the hydrolysate was re-separated by the same HPLC system. Negative ion mode FAB-LC/MS of the bioactive fractions of this hydrolysate revealed an [M-H] molecular ion peak at m/z 169 and 321. The 13C-NMR spectra were consistent with those of authentic gallic acid and galloyl gallic acid, a dimer of gallic acid. Authentic gallic acid also showed a triglyceride decreasing action, and galloyl gallic acid even more markedly decreased triglyceride. These findings suggest that the triglyceride-decreasing action of Japanese red pine needles is due to these polyphenol compounds. In addition, amino acids were also detected in the hydrolysate of PAC, indicating that the components in Japanese red pine needles that decrease triglycerides are complexes of a hydrolyzable tannin, which contains gallic acid and galloyl gallic acid as components and peptides. PMID- 9212986 TI - Uptake of methylchlorpromazine by brush-border membrane vesicles from rat small intestine. AB - The uptake of methylchlorpromazine (MCP), a quaternary derivative of chlorpromazine, was investigated using brush-border membrane vesicles isolated from rat small intestine. MCP was taken up rapidly by the vesicles, a major part of the uptake being due to binding to the membrane. Saturable MCP binding to the brush-border membrane was inhibited strongly by chlorpromazine, moderately by propantheline and imipramine, and slightly but significantly by methylbenactyzine and mepenzolate. However, choline and tetramethylammonium failed to exhibit any such inhibitory effect. The movement of MCP into the Intravesicular space was driven by an inside-negative transmembrane electrical potential difference (TEPD) induced by NaSCN or valinomycin. There was no effect of TEPD on MCP binding to the brush-border membrane. The data suggested that both rapid binding to the brush-border membrane and inside-negative TEPD, which is present physiologically across the membrane, play a significant role in the absorptive movements of MCP across intestinal epithelium. PMID- 9212987 TI - Microbial contamination of antiseptic-soaked cotton balls. AB - We investigated microbial contamination of in-use antiseptics at a hospital. No microbial contamination was observed in 70 samples of 0.02% benzalkonium chloride solution (500-ml volume), 70 samples of 1% titratable I2 povidone-iodine solution (250-ml volume), or 15 samples of 0.1% ethacridine lactate solution (500-ml volume) during use in reduced amounts. Nor was any microbial contamination observed in 70 samples of cotton balls soaked in 1% titratable I2 povidone-iodine solution in canisters or cotton gauze soaked in 70% (w/v) ethanol solution in canisters. However, among 70 samples of cotton balls soaked in 0.02% benzalkonium chloride solution in canisters, 6 (8.6%) were contaminated with 10(4) to 10(6) viable cells/ml. The microbial species detected were glucose non-fermentative bacilli such as Alcaligenes xylosoxidans and Pseudomonas putida. The contaminants obtained from cotton balls soaked in 0.02% benzalkonium chloride solution did not proliferate in that solution or in distilled water but showed rapid growth in the cotton balls soaked in either of these liquids. These findings suggested that benzalkonium chloride solution tends to become contaminated when cotton balls are immersed. Therefore, cotton balls soaked in benzalkonium chloride solution are not recommended as an antiseptic. When no other choice is available, the cotton balls should be soaked in benzalkonium chloride solution at the time of usage. PMID- 9212988 TI - Application of long-circulating liposomes to cancer photodynamic therapy. AB - Photodynamic therapy (PDT) as a cancer treatment is notable for its quite low side effects in comparison with those of chemotherapy and radiotherapy. However, the accumulation of porphyrin derivatives used in PDT into tumor tissues is rather low. Since long-circulating liposomes are known to accumulate passively into tumor tissues, we liposomalized a porphyrin derivative, benzoporphyrin derivative monoacid ring A (BPD-MA), and used these liposomes to investigate the usefulness of PDT for tumor-bearing mice. BPD-MA was liposomalized into glucuronate-modified liposomes, which are known to be long-circulating. These liposomes were injected i.v. into Balb/c mice bearing Meth A sarcoma, and tumor regression and survival time were monitored after irradiation with laser light. Tumor regression and complete curing of tumor (80% cure rate by the treatment with 6 mg/kg BPD-MA) were observed when long circulating liposomalized BPD-MA was injected and laser-irradiated. In contrast, only a 20% cure rate was obtained when the animals were treated with BPD-MA solution or BPD-MA entrapped in conventional liposomes. These results suggest that a long-circulating liposomal formulation of photo-sensitive agents is useful for PDT. PMID- 9212989 TI - Contribution of hydrophobicity of nonionic detergents to membrane lipid fluidity and disopyramide uptake by rat intestinal brush-border membrane vesicles. AB - The contribution of hydrophobicity of different types of detergents to disopyramide uptake by rat small intestinal brush-border membrane vesicles was studied in relation to their membrane lipid fluidity and the physicochemical parameters of the detergents, i.e., hydrophile-lipophile balance (HLB). Span-, Tween-type detergents or glycerol esters at non-solubilizing concentrations (0.01 0.05% (w/v)) decreased the extent of maximum uptake of the drug in the presence of outward H(+)-gradient, but not in the absence of the gradient. The fluorescence anisotropy of the vesicles using diphenylhexatriene (DPH), as reflected by its incorporation into the membrane inner lipid layer, decreased with the addition of all detergents used. In contrast, that of the vesicles using trimethylammoniumphenyl phenylhexatriene (TMA-DPH), which reflected its incorporation into the membrane outer lipid layer, increased depending on the concentration of Tween-type detergents except for Tween 81 and Tween 85, glycerol esters (MO-500, MO-750, ML-500 and ML-750); it decreased with the addition of Span-type detergents, Tween 81 and glycerol ester (MO-310). Therefore, the membrane lipid fluidity change of the outer leaflet, rather than the inner lipophilic domain, of the membrane vesicles caused by the detergents was found to be dependent on the hydrophobicity, but not on the type of detergent. This seems to correlate with the inhibitory effects on the facilitated uptake of the drug by the membrane vesicles. PMID- 9212990 TI - Kinetic characterization of binding and internalization of fractionated [3H]heparin in rat liver parenchymal cells in primary culture. AB - The binding and internalization of fractionated [3H]heparin (FH) was kinetically analyzed in rat liver parenchymal cells to clarify its cellular uptake mechanism. The binding of FH to the cell surface was saturable with the dissociation constant (Kd) of 53.5 nM and a maximum binding capacity (Bmax) of 19.9 pmol/mg protein. The binding of FH to the cell surface was competitively inhibited not only by heparan sulfate, a polyanion analogous to heparin, but also by rose bengal, an organic anion, suggesting the binding is based on an electric interaction requiring an anionic charge for substrates and consistent with the earlier suggestion of the involvement of the scavenger-like receptor. According to kinetic model analysis, the rate constants of association (K(on)), dissociation (k(off)), and internalization (k(int).app) were estimated to be 0.0005 nM-1 min-1, 0.0112 min-1, and 0.0056 min-1, respectively. Although both Kd and Bmax were larger than those reported in Kupffer cells, suggesting lower affinity and higher capacity in liver parenchymal cells, the apparent internalization rate constant was similar to that in Kupffer cells. We thus provided additional evidence suggesting that a scavenger-like receptor exists in rat liver parenchymal cells, and then kinetically characterized the surface binding and internalization of fractionated heparin by this receptor. PMID- 9212991 TI - Pharmacokinetic/pharmacodynamic analysis of neutrophil proliferation induced by recombinant granulocyte colony-stimulating factor (rhG-CSF): comparison between intravenous and subcutaneous administration. AB - Pharmacological effect after intravenous (i.v.) or subcutaneous (s.c.) administration of human recombinant granulocyte colony stimulating factor (rhG CSF) was evaluated by using a physiologically based pharmacokinetic/pharmacodynamic model. The increase of neutrophil counts in blood after s.c. administration of rhG-CSF (0.5-1.0 microgram/kg) was larger than that after i.v. administration of the same dose, while area under the curves of plasma concentration of rhG-CSF after s.c. administration was smaller than that after i.v. administration (Azuma et al., J. Clin. Therap. Med., 5, 1579-1603, 1989). Based on the pharmacokinetic/pharmacodynamic model considering metabolic turnover of neutrophil in vivo, time course of absolute neutrophil counts in blood after either type of rhG-CSF administration to normal healthy volunteers was analyzed. The nonlinear relationship between the concentration of rhG-CSF and in vitro activity for proliferation of neutrophil could be explained by the drug-receptor effector ternary complex model. These in vivo and in vitro models made it possible to understand the above-mentioned discrepancy of pharmacokinetic/pharmacodynamic behavior between i.v. and s.c. administration of rhG-CSF. Simulation of the neutrophil count-time profiles after repeated i.v. and s.c. dosing of rhG-CSF according to these models showed good agreement with the observed data. In order to obtain the rational dosage regimen of rhG-CSF from pharmacological and economical points of view, a slow constant infusion method may be more useful than rapid infusion. PMID- 9212992 TI - Determination of the functional domain of a mouse autonomous replicating sequence. AB - We previously isolated from mouse cells an autonomous replicating sequence (ARS) ARS65 (Ariga, Itani and Iguchi-Ariga, Mol. Cell. Biol. 7, 1-6, 1987). Here we report the nucleotide sequence of ARS65. The sequence from BgIII to EcoRI sites cloned as ARS was 2658 bp long. There exist three interesting domains: a TA repeat, a myc like box (essential sequence for c-myc ARS), and a T rich region. Cloned DNAs containing various segments of pARS65 were transfected to rat 3Y1 cells together with the hygromycinB resistance expression vector, and hygromycinB resistant clones were isolated. Established cell lines transfected with plasmids carrying either a myc-like box or a T rich region harbored the replicated plasmids, indicating that these two elements are necessary for the ARS function of pARS65. PMID- 9212993 TI - Z-100, extracted from Mycobacterium tuberculosis strain Aoyama B, inhibits the development of collagen-induced arthritis in mice. AB - We evaluated the effects of Z-100, extracted from human type Mycobacterium tuberculosis strain Aoyama B, on collagen-induced arthritis (CIA) in mice. One hundred thirty-five DBA/1J mice, 8 weeks of age, were assigned to 9 groups and immunized with bovine type II collagen (CII) or CFA. From the next day, Z-100 at doses of 0.004, 0.04, or 0.4 mg/kg B.W./d for 48 d was intradermally injected into the tail base. Methotrexate (MTX) at daily doses of 0.1, 0.3, or 1.0 mg/kg B.W. and cyclophosphamide (CY) at a daily dose of 5 mg/kg B.W. were used as reference drugs. The effects of these drugs on CIA mice were evaluated in terms of the incidence of CIA, the arthritis index (AI), and hind paw edema, after which the animals were sacrificed at 49 d, and both anti-CII antibody titer and delayed-type hypersensitivity (DTH) reaction were measured. In the arthritic control groups, the AI and hind paw edema were significantly increased after the second immunization on day 28. The anti-CII antibody titer and DTH reaction were significantly increased compared to normal mice on day 49. Z-100 significantly inhibited the AI at a dose of 0.4 mg/kg/d on day 49, and suppressed the incidence of both CIA and hind paw edema. Increases in both anti-CII antibody titer and DTH reaction in CIA mice were prevented by treatment with Z-100 at 0.4 mg/kg/d. MTX, in a dose-dependent manner, and CY, at a dose of 5 mg/kg/d, inhibited the incidence of CIA, AI, hind paw edema, anti-CII antibody titer and DTH reaction in CIA mice. Z-100 at a dose of 0.4 mg/kg was as effective as MTX was at a dose of 0.3 mg/kg against the DTH reaction, and it had no side effects. These results suggest the usefulness of Z-100 in patients with chronic rheumatoid arthritis. PMID- 9212994 TI - In vitro cytotoxicity of imidazolyl-1,3,5-triazine derivatives. AB - We examined in vitro cytotoxic activity of imidazolyl-1,3,5-triazine derivatives using human breast cancer cell lines (MCF-7, R-27, T-47D and ZR-75-1) and murine leukemia cell line (P388). The percentage of viable cells was determined by the 3 (4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazorium bromide (MTT) assay. Hexamethylmelamine (HMM), a 1,3,5-triazine derivative has previously been recognized as an antitumor agent effective against lung, ovarian and breast cancer, but failed to show a significant cytotoxic activity in the present study. In contrast, four imidazolyl-1,3,5-triazine derivatives, 2-(1-imidazolyl)-4,6 bis(morpholino)-1,3,5-triazine, 2-(1-imidazolyl)-4-morpholino-6-(3-thiazolidinyl) 1,3,5-triazine, 2-(4-cyano-4-phenylpiperidino)-4-(1-imidazolyl)-6-morpholino 1,3,5-triaz ine and 2-(1-imidazolyl)-4-(N-methyl-N-phenylamino)-6-morpholino 1,3,5-triazine showed cytotoxic activity for most cell lines, which was significantly greater than the activity of hydroxymethylpentamethylmelamine (HMPMM), a major metabolite of HMM. PMID- 9212995 TI - Absorption characteristics of azasetron from rectal and oral routes in rabbits. AB - The absorption characteristics of azasetron, a serotonin type 3 (5-HT3) receptor antagonist which is used for the treatment of chemotherapy-induced emesis and nausea, were investigated in rabbits. The serum concentrations of azasetron following rectal administration as a suppository increased rapidly and showed the mean tmax value of 0.18 h. The concentrations were greater after rectal administration than those after oral administration. The absolute bioavailability was significantly different between the rectal, 52.9% and oral doses, 21.6%. The mean Cmax and tmax values after the rectal dose were 904.8 ng/ml and 0.18 h, respectively, whereas those after the oral dose averaged 124.7 ng/ml and 0.85 h, respectively. These results indicate that azasetron is absorbed to a greater extent and more rapidly into the systemic circulation via the rectum than via the intestine in rabbits. Consequently, the suppository form of azasetron hydrochloride may be feasible for the treatment of chemotherapy-induced acute emesis and nausea. PMID- 9212996 TI - Synthesis and application of neoglycolipids for liposome modification. AB - We synthesized various glycolipid derivatives and examined the in vivo behaviors of liposomes modified with these novel glycolipid derivatives. Gal-t-psa (1,?8-(2 hexadecyloctadecanoylamido)-3,6-dioxaoctyl?-beta-D- galactoside), Lac-t-psa (3, 8 (2-hexadecyloctadecanoylamido)-3,6-dioxaoctyl beta-D-lactoside) and GalNAc-t-psa (4, 8-(2-hexadecyloctadecanoylamido)-3,6-dioxaoctyl 2-acetamido-beta-D galactopyranoside) modified liposomes were recognized by the liver. Lac-t-psa (3) modified liposome was accumulated to the highest degree, followed by GalNAc-t-psa (4) modified liposome and then Gal-t-psa (1) modified liposome. The intrahepatic distributions of Gal-t-psa (1), GalNAc-t-psa (4), Glc-t-psa (2, 8-(2 hexadecyloctadecanoylamido)-3,6-dioxaoctyl beta-D-glucopyranoside) and Lac-t-psa (3) modified liposomes were investigated. GalNAc-t-psa (4) and Lac-t-psa (3) modified liposome were accumulated to greater extents than Gal-t-psa (1) modified liposome in hepatic parenchymal cells. The intrahepatic distribution of these liposomes showed that Lac-t-psa (3) and GalNAc-t-psa (4) were preferable to Gal-t psa (1) for the selective delivery of liposomes to hepatic parenchymal cells. PMID- 9212997 TI - Application of glutaraldehyde-crosslinked chitosan as a scaffold for hepatocyte attachment. AB - The effectiveness of chitosan, a biocompatible polymer derived by the deacetylation of chitin, as a scaffold of hepatocyte attachment, was examined. Since chitosan gel was too fragile to use for cell culture, its free amino groups were crosslinked by glutaraldehyde to increase its strength. Rat hepatocytes seeded onto glutaraldehyde-crosslinked chitosan (GA-chitosan) gel could stably attach to the surface, retaining its spherical form, the same as in vivo, and then release a very small amount of lactate dehydrogenase during the 5 d culture period. By contrast, hepatocytes on a collagen-coated surface spread flat, and they released much more lactate dehydrogenase than those on the GA-chitosan gel. Hepatocytes on GA-chitosan also retained higher urea synthesis activity, a liver specific function, than those on the collagen-coated surface. These results indicate that chitosan is a promising biopolymer as a scaffold of hepatocyte attachment, which can be applied to an effective bioartificial liver support system. PMID- 9212998 TI - Cloning and bacterial expression of a gene encoding ribosome-inactivating proteins, karasurin-A and karasurin-C, from Trichosanthes kirilowii var. japonica. AB - A genomic DNA clone of karasurin was isolated using the polymerase chain reaction from Trichosanthes kirilowii var. japonica (Cucurbitaceae). The amino acid sequence deduced from the nucleotide sequence was consistent with previously reported sequences of karasurin-A and karasurin-C except for a putative signal peptide and extra amino acids at the C-terminus, neither of which is present in the natural protein. Recombinant karasurin was synthesized in Escherichia coli, in which the cloned karasurin gene was expressed under the control of the trc promoter. PMID- 9212999 TI - A novel method for induction and detection of anaphylactic reaction using the mouse abdominal wall (AW method). AB - We found that an antigen-specific anaphylaxis was induced by antigen challenge to the abdominal wall, ear auricle, or subcutaneous tissue in mice sensitized 9 days previously with antigen and adjuvant. The anaphylactic reaction was detected by vascular permeability at the injected site 7 minutes after challenge, which was the best time for estimation. A novel method (AW method) for induction and detection of the anaphylactic reaction in mice was established using the abdominal wall as the challenge site. This method could detect the anaphylactic response in mice 1 to 3 weeks after sensitization. The increase in vascular permeability was completely inhibited by administration of diphenhydramine. PMID- 9213000 TI - Inhibitory mechanisms of oleanolic acid 3-O-monodesmosides on glucose absorption in rats. AB - We examined the action mechanism of oleanolic acid 3-O-monodesmoside, momordin Ic (1), and oleanolic acid 3-O-glucuronide (2) for the inhibitory effect on the increase in serum glucose levels in oral glucose-loaded rats. Although 1 and 2 dose-dependently inhibited the increase in serum glucose levels in oral glucose loaded rats, these compounds showed no significant effects on serum glucose levels in normal rats, intraperitoneal glucose-loaded rats, and alloxane-induced diabetic mice. Furthermore, 1 and 2 were found to suppress gastric emptying in rats, and also to inhibit the glucose uptake in rat small intestine concentration dependently in vitro. These results indicate that 1 and 2 given orally have neither insulin-like activity nor insulin releasing-activity. 1 and 2 apparently inhibited glucose absorption by suppressing the transfer of glucose from the stomach to the small intestine and by inhibiting the glucose transport system at the small intestinal brush border. PMID- 9213001 TI - Identification and enrichment of human osteoprogenitor cells by using differentiation stage-specific monoclonal antibodies. AB - A major problem in developmental bone biology is the inability to clearly identify early progenitor cells of the osteogenic and related lineages. Identification of these cells is important for the study of their normal development and for determination of potential changes in skeletal diseases. The objective of the present study was to obtain specific markers for early progenitor cells. Monoclonal antibodies were raised against human marrow stromal fibroblastic cell cultures, known to be rich in progenitors for the stromal lineages. Antibodies were selected initially by their reactivity with these marrow cultures and their immunohistochemical localization in human fetal tissues, in progenitor cell regions adjacent to osteoblastic cells. Antibody HOP 26 was strongly reactive with cells in marrow stromal colonies at early stages of differentiation, before the induction of alkaline phosphatase activity, and decreased dramatically after the cells reached confluence. In sections of human fetal limb, binding of HOP-26 was restricted to cells in close proximity to the developing bone, in periosteum, and between the developing bone trabeculae. In adult trabecular bone tissue, HOP-26 was reactive with occasional cells present within the marrow spaces with osteoblasts, adipocytes, and fibrous tissue unreactive. No antibody binding was detected in sections of skin, muscle, appendix, brain, tonsil, or liposarcoma, or cultured SaOS II, MG63, or skin cells. In primary cell suspensions, HOP-26 was unreactive with blood cells but strongly reactive with 0.59 +/- 0.27% of nucleated marrow cells. The antigen associated with these cells was detectable both intracellularly and on the cell surface, and by using immunopanning, HOP-26 selected the marrow stromal fibroblastic colony-forming units (CFU-F). HOP-26 provides the means to identify osteogenic progenitor cells directly and with high specificity. The present studies demonstrate the value of this antibody in providing enriched populations of progenitor cells for experimental studies of osteogenic differentiation and in histopathology. PMID- 9213002 TI - Effect of rhBMP-2 on the osteogenic potential of bone marrow stromal cells from an osteogenesis imperfecta mouse (oim). AB - To understand whether osteogenesis imperfecta (OI) could result from defective differentiation of osteoprogenitor cells, we investigated the osteogenic potential of bone marrow stromal cells from a mouse model of human OI (oim). Bone marrow was flushed from the femurs and tibias of oim and normal littermates using a syringe with Dulbecco's modified Eagle's medium, and cells were allowed to adhere to flasks. Adherent cells were trypsinized and passaged weekly at a 1:4 split. The established stromal cells were assessed for collagen synthesis, alkaline phosphatase, and osteocalcin production in the presence or absence of rhBMP-2. The stromal cells were also assessed for mineralization by Von-Kossa staining and for exogenous gene transfer using adeno-lacZ and a retroviral vector. The bone marrow stromal cells from oim mice synthesized alpha 1(I) homotrimers as expected, whereas the stromal cells from the normal littermates synthesized alpha 1(I)2 alpha 2(I) heterotrimers. The bone marrow stromal cells exhibited low levels of alkaline phosphatase activity under basal conditions: upon treatment with rhBMP-2, the level of the alkaline phosphatase activity increased approximately 40-fold. Cytochemical staining of the cells confirmed the expression of alkaline phosphatase by the oim stromal cells and its augmentation by rhBMP-2. Osteocalcin production in the stromal cells was also enhanced approximately threefold by rhBMP-2. oim stromal cells grown in the presence of beta-glycerophosphate and ascorbic acid demonstrated Von-Kossa-positive solid deposits after 3 weeks in culture. Ten days after infection with adeno-lacZ, approximately 70% of oim stromal cells expressed the transgene product, and after infection with a retrovirus, approximately 20% of the cells expressed the transgene. These data indicate that bone marrow stromal cells, have osteogenic potential, and also the potential to be transduced with exogenous genes. Under basal conditions, however, the stromal cells from oim mice exhibited significantly lower levels of alkaline phosphatase activity than their normal littermates. PMID- 9213003 TI - Interleukin-1 beta enhances and tumor necrosis factor-alpha inhibits bone morphogenetic protein-2-induced alkaline phosphatase activity in MC3T3-E1 osteoblastic cells. AB - The modulatory effects of interleukin (IL)-1 beta and tumor necrosis factor (TNF) alpha on bone morphogenetic protein (BMP)-2- and -4-induced alkaline phosphatase (ALP) activity were examined in cultures of mouse MC3T3-E1 osteoblastic cells. Both BMP-2 and -4 significantly induced ALP in these cells. IL-1 beta alone had no effect on ALP activity, but it significantly enhanced BMP-2- and -4-induced ALP activity. TNF-alpha suppressed the induction of ALP by BMP-2 or -4. The results suggest that the action of BMP on osteogenic differentiation may be regulated by such immuno/inflammatory cytokines as IL-1 beta and TNF-alpha. PMID- 9213004 TI - Expression and localization of bone morphogenetic proteins (BMPs) and BMP receptors in ossification of the ligamentum flavum. AB - To clarify the pathogenesis of ossification of the ligamentum flavum (OLF), we examined the expression and localization of bone morphogenetic proteins (BMPs) and their receptors (BMPRs) in the ligamentum flavum of the patients with OLF by immunohistochemical staining and compared them with staining patterns in control patients. The BMPRs appeared extensively in mature and immature chondrocytes around the calcified zone and in spindle-shaped cells and round cells in the remote part from ossified foci in examined tissue of OLF. The ligands for BMPRs, BMP-2/-4 and osteogenic protein-1 (OP-1)/BMP-7, colocalized in OLF patients. In the control cases, expression of BMPs and BMPRs was observed around the calcified zone at the insertion of the ligamentum flavum to the bone, and limited expression was found in the smaller range. Thus, the expression profile of BMPs and BMPRs in OLF patients was entirely different from the control patients, suggesting that BMPs may be involved in promoting endochondral ossification at ectopic ossification sites in OLF, and that ossification activity is continuous in these patients. PMID- 9213005 TI - Rat tibial osteoblasts III: propagation in vitro is accompanied by enhancement of osteoblast phenotype. AB - Postproliferative confluent cultures of primary rat tibial osteoblasts (ROB), cultured in medium supplemented with ascorbic acid and beta-glycerophosphate (AS bGP, differentiation medium) express, in sequence, specific bone markers which identify a succession of maturation stages, and eventually form mineralized noduli. We report an investigation on the effect of extensive proliferation in vitro in unsupplemented medium on the osteogenic potential of mass cultures of ROB. The growth rates of the populations, derived from two independent primary cultures, was constant throughout 110 cumulative population doublings (CPD) in culture. Propagated cells maintained features similar to osteoblasts in primary cultures with respect to serum and anchorage dependence for growth and to the chemokinetic effect on endothelial cells exerted by their conditioned media (CM). Propagated populations, set at confluence in differentiation medium, were tested for the expression of early [alkaline phosphatase (AP)] and late [osteocalcin (OC); bone sialoprotein (BSP); 45Ca incorporation and mineralization] osteogenic markers. We observed an increase, parallel to the increase in CPD, in both the level of maximal expression of AP (enzyme/microgram cellular DNA) and in the frequency of nodules, reaching five- to sixfold (at 78 CPD) and eightfold (at 60 CPD), respectively, the levels of primary cultures. AP expression (enzyme and mRNA) persisted during mineralization and 45Ca incorporation. The time required by propagated cultures for the formation of nodules decreased with increase of CPD, and was reduced to less than one third at 87 CDP. Nodules became mineralized over a similar lapse of time as in primary cultures and were positive by histochemistry for BSP and OC. We also obtained osteogenic clones from two independent cultures after 72 CPD. 90% of these showed an osteoblast phenotype, expressing AP and forming nodules positive for OC and BSP, which mineralized. Timing of formation and frequency of nodules/plated cells in clones was similar to that found in propagated cultures of equivalent CPD. In summary, propagated ROB populations and derived clones showed enhanced osteoblast phenotype, possibly due to an increase in osteogenic cells and enrichment of proliferating mature osteoblasts, consequent to extended propagation in culture. PMID- 9213006 TI - 1 alpha,25-dihydroxyvitamin D3 stimulates sodium-dependent phosphate transport in osteoblast-like cells. AB - 1 alpha,25-Dihydroxyvitamin D3 [1,25(OH)2D3] stimulates calcium and phosphate absorption in the duodenum. Because osteoblastic cells appear to be responsible for Pi deposition in bone, we analyzed the effect of 1,25(OH)2D3 on sodium dependent phosphate (NadPi) transport in newborn rat calvaria-derived osteoblasts and in a rat bone-derived cell line, PyMS. 1,25(OH)2D3 at 10(-9) mol/L stimulated NadPi uptake 1.6-fold in PyMS and 1.8-fold in calvaria cells after 24 h, but not after 2 h and 6 h. Insulin-like growth factor (IGF)-I stimulated NadPi transport 1.8-fold in both cell types within 2 h. There was no change in Na-dependent alanine transport or in Na-independent uptake of Pi and alanine, and the effects of 1,25(OH)2D3 on NadPi transport were not associated with corresponding changes in cell number, protein content, or alkaline phosphatase activity. Actinomycin D, an inhibitor of transcription, prevented the stimulatory effect of 1,25(OH)2D3 (but not that of IGF-I) on NadPi transport; stimulation of NadPi transport by 1,25(OH)2D3 may depend on an increased de novo synthesis of transporters. Our studies confirm that IGF-I stimulates NadPi uptake and show, for the first time, a specific stimulatory effect of 1,25(OH)2D3 on NadPi transport in osteoblasts in vitro. PMID- 9213008 TI - Saccharated ferric oxide (SFO)-induced osteomalacia: in vitro inhibition by SFO of bone formation and 1,25-dihydroxy-vitamin D production in renal tubules. AB - A 60-year-old man with portal hypertensive gastropathy due to type C liver cirrhosis developed severe bone pains, marked hypophosphatemia with inappropriately increased urinary excretion of phosphate (%TRP; 9.6%), and hyperalkaline phosphatasia, after intravenous administration of saccharated ferric oxide (SFO) at a dose of 80-240 mg/week over a period of more than 5 years. The total iron infused was estimated to be more than 25 g. On a diagnosis of SFO-induced osteomalacia, the infusion of iron was immediately discontinued, and phosphate and vitamin D2 (1000 IU/day) were administered. Serum levels of 25 OHD2 increased after 1 week, whereas levels of 1,25-(OH)2D2 did not increase until 3 months later, accompanied by improvement of renal tubular reabsorption of phosphate and gradual improvement of the bone pains. The patient has been doing well for the last 2 years, with normal serum levels of phosphate, calcium, and alkaline phosphatase, without any supplementation of phosphate, vitamin D, or iron-containing agents. In primary culture of neonatal mouse renal tubules, in which 1,25-(OH)2D3 was produced from 25-OHD3 in response to PTH, SFO significantly inhibited PTH-induced production of 1,25-(OH)2D3 at 30 mumol/L, which is attainable in the urine of patients receiving a therapeutic intravenous dose of SFO. Furthermore, SFO decreased the calcium content and inhibited 45Ca incorporation in cultured fetal mouse parietal bones at 3 mumol/L. Such SFO concentration may be transiently observed in the plasma of patients receiving excessive intravenous doses of SFO for a prolonged period. These in vitro findings together with the clinical observations suggest that SFO, after filtration through the glomerulus and reabsorption in the proximal renal tubules, impaired proximal renal tubular function, such as tubular reabsorption of phosphate and 1 alpha-hydroxylase activity, leading to hypophosphatemic osteomalacia. Furthermore, it is highly likely that SFO in the peripheral blood, when transferrin is saturated with iron, may impair bone formation and aggravate osteomalacia. Although SFO-induced osteomalacia is reversible simply by discontinuation of the agent, excessive and prolonged administration of SFO should be avoided. PMID- 9213007 TI - Absence of androgen-mediated transcriptional effects in osteoblastic cells despite presence of androgen receptors. AB - Androgen excess and deficiency affect skeletal maturation and bone cell function. Understanding the molecular basis for these androgen effects could improve therapy/prevention of short stature and osteoporosis. Androgens act through binding to androgen receptors (ARs), which modulate gene transcription via interactions with DNA response elements on target genes. Because osteoblasts contain ARs at levels just below certain androgen-sensitive tissues, we sought to define the function of AR in a number of commonly used osteoblastic cell lines. Presence and quantification of AR protein and mRNA were evaluated by ligand binding assay, western blotting, and RNAse protection assay. AR-containing osteoblastic cell lines were exposed to nonaromatizable androgens and effects on gene expression were assessed. We found no evidence for direct effects of androgen on endogenous genes nor was androgen involved in modulation of parathyroid hormone effects on early gene activation. Androgen-sensitive reporter gene constructs were stimulated by androgen only when AR cDNA expression vectors were introduced into cells by cotransfection. We conclude that, in commonly used osteoblastic cell lines, the presence of AR at the levels described here does not guarantee androgen transcriptional activity. The effects of androgen on bone in vivo may involve direct stimulation of osteoblastic cells in a different setting or stage of differentiation. Alternatively, androgen may act on bone cells other than osteoblasts, or through metabolic conversion to estrogens. PMID- 9213009 TI - Alendronate prevents cyclosporin A-induced osteopenia in the rat. AB - Post-transplantation bone disease is an increasingly recognized clinical entity whose etiology is multifactorial. The immunosuppressant agent cyclosporine-A (CsA) has repeatedly been shown experimentally to induce a high-turnover osteopenic state. Alendronate (Alen.) is a new generation bisphosphonate having far greater antiresorptive potency than previous bisphosphonates. It inhibits osteoclast resorption in vitro and in vivo without adversely affecting bone mineralization. This study was designed to investigate whether alendronate could prevent CsA-induced osteopenia in the rat. Forty-eight 8-month-old male Sprague Dawley rats were randomized into four groups to receive the following for 28 days: (1) CsA vehicle (veh.) p.o. daily and alendronate vehicle subcutaneously (s.c.) twice/week, (2) CsA 15 mg/kg p.o. daily and Alen. veh. s.c. twice/week, (3) Alen. 70 micrograms/kg s.c. twice/ week and CsA veh. p.o. daily, and (4) CsA 15 mg/kg p.o. daily and Alen. 70 micrograms/kg s.c. twice/week. Rats were weighed and bled and serum was assayed serially for calcium, PTH, 1,25(OH)2vit.D, and osteocalcin. Tibiae were removed following sacrifice on day 28, after double demeclocycline and calcein labeling, for histomorphometric analysis. Treated groups were compared to the vehicle-treated control. We confirmed previous findings that CsA produces elevated 1,25(OH)2 vitamin D and serum osteocalcin levels. Alendronate treatment by itself decreased osteocalcin by day 28 and resulted in a marginal decrease in serum total calcium on day 14. The histomorphometry findings reconfirmed that the administration of CsA induces a state of high-turnover osteopenia. Alendronate prevented CsA's adverse effects, particularly in maintaining trabecular bone volume, presumably by decreasing bone remodeling. Alendronate would seem to hold therapeutic promise in post transplantation bone disease. PMID- 9213010 TI - Short-term immobilization-induced cancellous bone loss is limited to regions undergoing high turnover and/or modeling in mature rats. AB - Estrogen and calcium deficiencies increase both bone resorption and formation, whereas immobilization mainly decreases bone formation. How these functionally different risk factors for bone loss interact in cancellous bone undergoing modeling or remodeling activity is not well understood. Mature (6-month-old) female rats were subjected to sham operation (sham), ovariectomy (ovx), dietary calcium deficiency (LoCa, 0.1% Ca), and sciatic and femoral denervation (IM), ovx+IM, or LoCa+IM for 4 weeks. The primary spongiosa, the region of active modeling within 1 mm of the growth plate, in ovx, LoCa, and IM groups showed a decrease in cancellous bone volume, trabecular number, and connectivity when compared to sham controls. Groups combining two risk factors exhibited additive changes when compared with single risk factor groups. In the secondary spongiosa, an area with little modeling activity, ovx and LoCa groups, as expected, lost bone. In contrast with the primary spongiosa, IM alone did not induce bone loss in the secondary spongiosa, and the groups with a combination of IM and ovx or IM and LoCa showed a greater bone loss than either ovx or LoCa alone. Ovx and LoCa groups showed increases in both bone formation rate and eroded surface in the secondary spongiosa, while IM groups showed a decrease in bone formation rate. Combining IM with either ovx or LoCa resulted in increased eroded surface. The effects on cortical bone were assessed at the tibio-fibular junction. A trend toward decreased percentage of cortical bone area and an increase in marrow cavity area were observed in the combined deficiency groups only. These changes were the result of a statistically significant increase in endosteal eroded surface in IM+ovx and IM+LoCa groups. Our results demonstrate that immobilization induced bone loss is restricted to the primary spongiosa where most modeling events occur. However, the inhibitory effect of IM on bone formation in the secondary spongiosa is unmasked in remodeling sites when a high turnover state is provided by either estrogen or dietary calcium deficiency. These results suggest that the presence of a risk factor, such as immobilization, which in the short term causes inhibition of bone formation, does not predispose the skeleton to rapid cancellous bone loss except when accompanied by modeling or high turnover. PMID- 9213011 TI - Accelerated bone mineral loss following a hip fracture: a prospective longitudinal study. AB - The purpose of this prospective study was to monitor the bone mineral density (BMD) of the lumbar spine and contralateral femoral neck in the first year following an osteoporosis-related fracture of the hip. Eighty-three elderly patients (mean age 77 years) who had sustained a hip fracture had determinations of BMD made at the time of fracture; 49 of these patients were available for reassessment of BMD 1 year later. The change in BMD was correlated with pre- and postinjury variables, such as ambulatory ability, dietary intake of calcium, serum vitamin D levels, mental status, and routine serologies. The mean decrease in BMD in the year following fracture was 5.4% from the contralateral femoral neck and 2.4% from the lumbar spine. Calcium intake correlated with the loss of BMD from the femoral neck (p = 0.015), but not the lumbar spine. Patients with daily calcium intakes of less than 500 mg/day had a more than 10% decrease in femoral neck BMD in the year following their hip fracture. Serum 1,25-dihydroxy vitamin D level correlated with loss of MBD from the lumbar spine (p = 0.001), but not from the femoral neck. There was no correlation between the loss of bone mineral from either measurement site and age, sex, level of ambulation, or mental status. The loss of BMD from the femoral neck in the year following a hip fracture is more than five times that reported in the nonfractured population. This accelerated rate of loss can have drastic consequences in an elderly population already exhibiting osteopenia and propensity to fall. Investigation of pharmacologic or other interventions in the first critical year following a hip fracture may potentially blunt this accelerated rate of bone loss and lessen the risk of subsequent fractures. PMID- 9213012 TI - Circadian variation in bone resorption is not related to serum cortisol. AB - Serum osteocalcin, serum procollagen type I carboxyterminal propeptide (sPICP), and the urinary excretion of pyridinium crosslinks (biochemical markers of bone formation and resorption) all exhibit a circadian variation with a peak during the night. This study was performed to investigate the influence of the endogenous circadian rhythm in cortisol on the biochemical markers of bone turnover. Participants included 11 patients substituted with hydrocortisone due to either hypopituitarism (n = 7) or bilateral adrenalectomy (n = 4). Their daily tablet intake of hydrocortisone was divided in four equal doses in order to abrogate the known circadian variation in cortisol. 24 healthy postmenopausal women served as controls. The study design was performed over 24 h, with blood samples taken every 3 h, and urine collected in 3 h aliquots. Urinary pyridinium crosslinks (Pyr/ Cr, D-Pyr/Cr), serum osteocalcin (sOC), and serum PICP were measured. Patients without a circadian variation in cortisol had normal circadian variation in the urinary excretion of pyridinium crosslinks and sPICP, but no circadian rhythm in serum osteocalcin. We conclude that the etiology of the circadian rhythm in the biochemical markers of bone turnover is still unknown. This study indicates that the circadian variation in sOC can be controlled by the endogenous circadian variation in serum cortisol, whereas this hormone does not control the circadian variation in either the serum PICP or the urinary excretion in pyridinium crosslinks. PMID- 9213013 TI - Polymorphisms of the interleukin-6 gene are associated with bone mineral density. AB - Genetic factors play an important role in determining bone mass and several genes probably act as regulators of this process. Interleukin-6 (IL-6) is a candidate gene for regulation of bone density, since it has stimulatory effects on cells of the osteoclast lineage and has been implicated in the pathogenesis of bone loss associated with estrogen deficiency. Here we studied the relationship between bone mineral density (BMD) and a polymorphic AT rich minisatellite repeat in the 3' flank of the IL-6 gene in a cohort of 200 women. Six length variants were identified (designated A-F), but four of these were rare such that the last majority of individuals fell into one of two common genotypes: F/F (58.5%) and C/F (27.5%). There was a significant relationship between IL-6 genotype and bone mass at the lumbar spine as determined by analysis of variance (p = 0.04) and a similar trend for bone mass at the femoral neck (p = 0.11). When BMD values were compared in the two common genotypes, we found that spine BMD values were significantly higher in the C/F genotype (mean +/- SEM = 0.94 +/- 0.04 g/cm2) as compared with the F/F genotype (0.81 +/- g/cm2; p = 0.012). A similar trend was seen for hip BMD values, but here, the difference failed to reach statistical significance (p = 0.06). Further analysis showed that genotype-specific effects on bone mass were observed in both premenopausal and postmenopausal women and did not increase with age, suggesting that the association between IL-6 polymorphisms and bone density may be mediated by an effect on peak bone mass, rather than rate of bone loss. We conclude that bone mass is associated with two common polymorphisms of the IL-6 gene. Although the mechanisms that underlie this association will require further research, our data suggest that polymorphic variation at the IL-6 gene locus may contribute to the genetic regulation of bone mass. PMID- 9213014 TI - Primary hyperparathyroidism: biochemical markers and bone mineral density at multiple skeletal sites in Danish patients. AB - Biochemical bone markers and bone mineral density (BMD) in spine, hip, and forearm were measured, before surgery, in 30 patients with mild to moderate primary hyperparathyroidism (PHP) (25 women and 5 men; mean age 54 +/- 12 years, range 26-73 years) and compared with normal controls. A group of 291 healthy adults (181 women and 110 men) served as controls for BMD. A smaller group of 30 normal individuals (25 women and 5 men; mean age 54 +/- 12 years; range 26-74 years) were used as matched normal controls. Parameters of bone formation (s osteocalcin, s-alkaline phosphatase activity, and s-bone isoenzyme alkaline phosphatase activity) and bone resorption (s-type-1 collagen telopeptide) were considerably increased in patients with PHP compared with normal controls (p < 0.01 for all parameters). BMD was found to be reduced in the hip (trochanteric: 95.1 +/- 14.7% of expected, p < 0.05; intertrochanteric: 95.2 +/- 13.8% of expected, p < 0.05), and the forearm (proximal: 93.3 +/- 12.2% of expected, p < 0.05; mid: 91.8 +/- 11.6% of expected, p < 0.001; distal: 90.7 +/- 13.1% of expected, p < 0.001). Spine BMD was found significantly reduced in premenopausal (87.8 +/- 7.6% of expected, p < 0.05) but not in postmenopausal patients, and although normal women showed a decrease in spinal BMD with increasing age this was not found in the PHP women. Forearm BMD was reduced in both pre- and postmenopausal patients (distal forearm: 86.7 +/- 12.2% of expected, p < 0.05; 87.6 +/- 12.1% of expected, p < 0.01, respectively). It was concluded that Danish patients with mild or moderate PHP have only small reductions in BMD. The bone loss is mainly found in the appendicular skeleton. PMID- 9213015 TI - Volumetric quantitative computed tomography of the proximal femur: precision and relation to bone strength. AB - We have developed a three-dimensional computed tomography (CT) scanning and image analysis method for measurement of trabecular and integral bone mineral density (BMD) and geometry in automatically determined femoral-neck and trochanteric subregions of the proximal femur. We measured the correlation of the density and geometry variables to femoral strength assessed in vitro under loading simulating a single-limb condition and a fall to the side. While BMD alone accounted for 48% 77% of the variability in strength for the stance loading configuration, femoral neck cross-sectional area (minCSA) and femoral neck axis length (FNAL) also contributed independently to femoral strength, and a combination of BMD and geometry variables explained 87%-93% of the variance in the data. For the fall loading configuration, trochanteric trabecular BMD alone explained 87% of the variability of strength. The reproducibility in vivo of the technique was assessed in a group of seven postmenopausal women, who underwent repeat scans with repositioning. For trabecular BMD, the precision was 1.1% and 0.6% for the femoral neck and trochanteric subregions, respectively, compared to 3.3% and 1.6% for the corresponding integral envelopes. Thus, trabecular BMD measurements were reproducible and highly correlated to biomechanical strength measurements. These results support further exploration of quantitative CT for assessment of osteoporosis at the proximal femur. PMID- 9213016 TI - Improved reproducibility of broadband ultrasound attenuation of the os calcis by using a specific region of interest. AB - Quantitative ultrasound (QUS) bone measurement is a promising, relatively new technique for the diagnosis of osteoporosis. Contrary to the more established method of bone densitometry (measurement of bone mineral density, BMD, e.g., using dual X-ray absorptiometry, DEXA), QUS does not employ ionizing radiation. It has, however, been a problem to achieve sufficient reproducibility of the QUS measurements. The aim of this study is to evaluate the possible advantages of measuring broadband ultrasound attenuation (BUA) at a region of interest (ROI) instead of at a fixed position, in terms of in vivo precision and correlation to hip bone mineral density. BUA was measured in 27 premenopausal women, 28 postmenopausal women, an 22 men on the DTU-one. Using high resolution images, a ROI is defined in the posterior part of the os calcis as an area with a local minimum of attenuation and a fixed position within the os calcis is defined relative to the water bath. All BUA measurements were performed twice. BMD at the hip was measured on the QDR-2000. The mean BUA values were significantly different between pre- and postmenopausal women, p = 0.0001 for both the ROI (BUAROI) and the fixed position (BUAFIX). The ROI was found in all subjects and was readily reproducible. The precision at the ROI: 1.20 CV% (95% CI: 1.01-1.29 CV%) was significantly better than at the fixed position: 3.87 CV% (95% CI: 3.23 4.48 CV%). BUAROI (r = 0.64) correlated significantly better than BUAFIX (r = 0.35) with HIP BMD. In conclusion, the use of an imaging technique enables BUA measurements to be performed at a ROI. The precision of BUA at the ROI is significantly better than at the fixed position. BUA measured at the ROI correlates better with HIP BMD than BUA measured at the fixed position. PMID- 9213017 TI - Effects of anteversion on femoral bone mineral density and geometry measured by dual energy X-ray absorptiometry: a cadaver study. AB - The effect of femoral neck anteversion on bone mineral density (BMD) and geometry as measured by dual energy X-ray absorptiometry (DXA) was assessed using 64 right proximal femora from 36 male and 28 female cadavers. The anteversion angle was measured on computed tomography (CT) images, and DXA measurements were made both in the neutral position (i.e, at 0 degree anteversion, femoral neck axis parallel to the table) and in the simulated anteverted position (i.e., femoral shaft axis parallel to the table, greater and lesser trochanters in contact with the table, and femoral neck free). The mean anteversion angle measured by CT was 19.3 degrees (range 6 degrees-38 degrees). Anteversion was associated with a significant elevation in femoral neck BMD of +2.8% (range -5.3%-(+)9.8%) (p < 0.05), and the femoral neck BMD increased with increasing anteversion (p < 0.01). Trochanteric BMD was less affected by anteversion, with an average increase of only 0.2% (range -5%-5.9%) (p = n.s.) in the anteverted position, but there was a significant positive association between the change in trochanteric BMD and the anteversion angle (p < 0.01). Anteversion produced a mean reduction of -2.4% (range -7.6%-(+)4.3%) (p < 0.001) in apparent femoral neck axis length, while femoral neck width remained generally unaffected. These data confirm that femoral BMD as measured by DXA is affected by femoral anteversion with a lesser magnitude than previously reported. The use of trochanteric BMD may minimize the influence of anteversion. While the mean changes in BMD and neck axis length attributable to anteversion are modest, the considerable interindividual variability in the magnitude of the effects demonstrates that other factors, such as, the complex geometry of femoral neck modifies the effect of anteversion on BMD measurements. The error in BMD introduced femoral anteversion may represent a significant confounding influence in cross-sectional and longitudinal studies. Careful repositioning of the foot and leg is essential in monitoring changes in BMD longitudinally. Knowledge of the effects of femoral anteversion may assist in understanding the relation of femoral BMD and neck axis length to hip fracture. PMID- 9213019 TI - Local drug delivery. PMID- 9213018 TI - Tibial ultrasound velocity measured in situ predicts the material properties of tibial cortical bone. AB - Quantitative ultrasound (QUS) is currently being investigated as a possible alternative or adjunct to X-ray-based methods for assessing osteoporosis and fracture risk. It has been proposed that QUS may allow measurement of bone "quality," such as bone architecture or material properties. In this study, we used human cadaveric specimens to evaluate whether ultrasound velocity measurements performed in situ at the midtibia were correlated with the mechanical properties of tibial cortical bone. We obtained 26 human lower limbs (10 men and 16 women) with a mean (+/-SD) age of 81 +/- 12 and range of 53-98 years. The longitudinal ultrasound velocity of the cortical bone at the anteromedial midtibia (tUV, meters per second) was assessed in the intact legs (SoundScan 2000, Myriad Ultrasound, Rehovot, Israel). Then a cylinder of cortical bone was removed from the anterior tibia at the site of QUS scanning, scanned using peripheral quantitative computed tomography to determine bone density, and mechanically tested in tension to failure. We found that tUV of the intact legs correlated strongly with bone density of the cortical bone specimens (r2 = 0.74, p < 0.0001). Both bone density (r2 = 0.89, p < 0.0001) and tUV (r2 = 0.84, p < 0.0001) were very strongly correlated with the cortical bone elastic modulus. In addition, both tUV (r2 = 0.75) and bone density (r2 = 0.80) were highly correlated with the ultimate strength of the cortical bone specimens. In summary, tibial ultrasound velocity measured in situ correlated with the material properties of tibial cortical bone nearly as strongly as did bone density. (Bone 21:119-125; 1997). PMID- 9213020 TI - Local delivery of c-myb antisense oligonucleotides during balloon angioplasty. AB - Intraluminal delivery of antisense oligonucleotides to c-myb was assessed following balloon angioplasty in swine peripheral arteries. Successful delivery and intramural persistence of oligonucleotide for over 24 h were demonstrated following angioplasty with hydrogel balloons coated with 32P-labeled antisense. Delivery of fluorescein-labeled antisense demonstrated further localization within the arterial media and intracellularly. Preliminary in vitro studies demonstrated the feasibility of inhibition of porcine lymphocyte proliferation using the murine antisense to c-myb. Twelve iliac or carotid arteries underwent angioplasty with antisense-coated balloons, while the contralateral vessels underwent angioplasty with the same-sized balloons coated with the complementary sense strand. Six to seven days later, dilated arterial segments were surgically isolated. In 10 of 12 vessel pairs, antisense-treated vessels demonstrated less cellular proliferation than did contralateral sense-treated vessels, as assessed by quantitative immunohistochemical staining of proliferating cell nuclear antigen, and smooth muscle cell proliferation was reduced 18% in antisense treated vessels compared to the contralateral sense-treated vessels (PCNA positive nuclear area: 7.7 +/- 4.9% vs. 9.3 +/- 5.2%, P < 0.04)-intraluminal delivery of antisense oligonucleotides to c-myb is feasible with a catheter-based system and may reduce smooth muscle cell proliferation following arterial injury. PMID- 9213021 TI - Feasibility and efficacy of locally delivered enoxaparin via the Channeled Balloon catheter on smooth muscle cell proliferation following balloon injury in rabbits. AB - To determine the potential utility of local treatment of enoxaparin on restenosis, four groups of rabbits underwent balloon injury of bilateral iliac arteries with the Channeled Balloon (balloon/artery = 1.1), followed by assigned treatment (5 controls received local saline, 7 local-treatment rabbits received a one-time local delivery of 10 mg/kg of enoxaparin, 5 systemic-treatment rabbits received 10 mg/kg of enoxaparin subcutaneously once daily for 1 wk, and 5 combined-treatment rabbits received both local and systemic enoxaparin). The percentage of nuclei positive for proliferating cell nuclear antigen/microns2 of media 1 wk later was 1.97 +/- 2.01 for the control group, 2.68 +/- 2.52 for the local group, 0.22 +/- 0.32 for the systemic group, and 0.07 +/- 0.09 for the combined group (P < 0.0001 by Kruskal-Wallis test, with P < 0.05 for combined treatment group vs. controls or local treatment group and systemic vs. local groups). Feasibility study with 3H-enoxaparin showed intramural retention of 0.1 0.2% of locally delivered amount for 24 h. We conclude that one-time local delivery of enoxaparin following angioplasty is ineffective in inhibiting medial smooth muscle cell proliferation, most likely due to low efficiency. Only sustained treatment resulted in significant reduction. PMID- 9213022 TI - Site-specific intracoronary thrombolysis with urokinase-coated hydrogel balloons: acute and follow-up studies in 95 patients. AB - Conventional balloon angioplasty in the presence of intracoronary thrombus is associated with an elevated risk for acute myocardial infarction, emergency bypass surgery, and death. The purpose of this study was to assess the safety and efficacy of a new technique to treat thrombus-containing stenoses consisting of the local delivery of urokinase directly to the site of intraluminal clot with hydrogel-coated balloons. Ninety-five patients with angiographically apparent intracoronary thrombus were treated with urokinase-coated hydrogel balloons either prior to (n = 74) or following (n = 21) conventional balloon angioplasty. Clinical diagnoses for the study group included acute myocardial infarction in 50 patients, postinfarction angina in 23 patients, and unstable angina in 22 patients. All hydrogel balloons were initially coated with urokinase by immersing the inflated balloon in a concentrated Abbokinase solution (50,000 units/ml) for 60 s. All patients were subsequently treated with drug-coated balloons using a balloon:artery ratio of 1:1, a mean of 2.2 +/- 1.2 inflations, and a mean total inflation time of 7.5 +/- 4.9 min. Use of urokinase-coated balloons resulted in angiographic disappearance of intracoronary thrombus in 78 patients, improvement in 14, and no change in the remaining 3 patients. Following hydrogel balloon use for the entire 95 patients, TIMI flow increased from 1.4 +/- 1.2 to 2.9 +/- 0.4, minimal lumen diameter increased from 0.4 +/- 0.4 to 2.0 +/- 0.6 mm, and thrombus score decreased from 2.0 +/- 0.9 to 0.2 +/- 0.6 (all P < 0.01). Procedural and early in-hospital complications were noted in 7 of the 95 patients (7.4%) and included abrupt closure in 3 patients, distal embolization in 1 patient, no reflow in 1 patient, sidebranch occlusion in 1 patient, and late closure in 1 patient. Two of the 3 patients with abrupt closure and the single patient with late closure required intracoronary stenting to maintain vessel patency. Two of these 7 patients sustained small myocardial infarctions, although no patient required emergency bypass surgery or experienced a procedural death. Late clinical follow-up (mean = 8.3 +/- 6.6 months; range = 2 wk to 29 mo) demonstrated adverse recurrent events in 29 of the 95 patients (30.5%), including death (n = 5), myocardial infarction (n = 2), and recurrence of angina (n = 22). The results of this study suggest that intracoronary thrombolysis can be safely and rapidly achieved by using limited quantities of urokinase delivered directly to the site of intraluminal clot with hydrogel balloons. Use of this technique may result in improved acute outcomes in comparison with conventional techniques currently being used to treat thrombus-containing stenoses. PMID- 9213023 TI - Local urokinase delivery with the Channel balloon: device safety, pharmacokinetics of intracoronary drug delivery, and efficacy of thrombolysis. AB - The Channel balloon is a new local drug-delivery catheter that has the dual capability of high-pressure lesion dilation and low-pressure drug infusion. The purpose of this study was to assess the safety and efficacy of this device in the local delivery of urokinase in the porcine model. Three in vivo protocols were performed in 57 anesthetized swine to assess the safety of Channel balloon use in the coronary vasculature, the pharmacokinetics of local urokinase delivery, and the ability of the catheter to lyse intraluminal thrombus. First, safety studies were performed in 18 coronary vessels in 13 pigs to compare angiographic and histologic changes following use of the Channel balloon with conventional balloon angioplasty. Second, intramural deposition of 123I-labeled urokinase was measured in 24 coronary arteries in 20 pigs to assess the efficiency and technical determinants of urokinase delivery and the time course of intramural drug retention. Finally, an in vivo thrombus model was used in 24 pigs to compare the thrombolytic capacity of local urokinase delivery with the Channel balloon in comparison with conventional urokinase infusion techniques. All balloon inflations and drug infusions with the Channel balloon were well tolerated in all animals without adverse angiographic, hemodynamic, or electrical sequelae. Comparative histologic studies with the Channel balloon demonstrated no additional vessel trauma beyond that seen with conventional balloon angioplasty. Between 0.09 and 0.35% of infused urokinase was intramurally deposited, with intracoronary persistence for at least 5 h. Drug infusion pressure did not significantly affect drug deposition, although larger amounts of urokinase were deposited with larger balloon:artery ratios and higher urokinase concentrations. In comparison to conventional systemic and guiding catheter infusions, local delivery of urokinase with the Channel balloon resulted in higher levels of clot dissolution. These studies have demonstrated safe intracoronary use of the Channel balloon in the porcine model. Local infusion of urokinase with this device results in significant intramural drug deposition that persists for at least 5 h. In comparison with conventional thrombolytic techniques, local urokinase delivery with the Channel balloon may result in enhanced intravascular thrombolysis. PMID- 9213024 TI - Treatment of thrombotic saphenous vein bypass grafts using local urokinase infusion therapy with the Dispatch catheter. AB - Percutaneous treatment of thrombotic stenoses or total occlusions in aged saphenous vein bypass grafts is associated with a significant incidence of complications primarily related to distal embolization. The purpose of this study was to assess the efficacy of local urokinase delivery with the Dispatch catheter prior to balloon angioplasty and/or intragraft stent placement as a new technique of vein graft revascularization. Local urokinase delivery with the Dispatch catheter was performed in 15 saphenous vein grafts (mean age = 11.7 +/- 2.5 yr) in 13 patients with unstable or postinfarction angina. The target lesion was a total occlusion in 5 of the procedures and a severe vein graft stenosis in the remaining 10. In all cases, urokinase was administered directly to the site of the stenosis/occlusion via the Dispatch catheter at 0.5 cc/min and at a concentration of 30,000 units/cc. The mean urokinase infusion time for the 15 procedures was 33 +/- 10 min (range = 10-60 min) and the mean urokinase dose was 495,000 +/- 158,000 units (range = 150,000-900,000 units). Following Dispatch therapy, mean minimal lumen diameter increased from 0.34 +/- 0.32 to 1.81 +/- 0.78 mm (P < 0.01), mean TIMI flow increased from 1.9 +/- 1.4 to 2.8 +/- 0.8 (P < 0.06), and mean thrombus score was reduced from 2.3 +/- 0.6 to 0.3 +/- 0.8 (P < 0.01). Mild no reflow was noted in two cases, although no patient demonstrated angiographic evidence of gross distal embolization. One of the patients with no reflow also demonstrated a small increase in cardiac enzymes. Subsequent balloon angioplasty/stent placement was successful in 14 of the 15 procedures (93% success rate). This preliminary report suggests that pretreatment of thrombotic saphenous vein graft stenoses with local urokinase delivery via the Dispatch catheter may decrease intragraft thrombus and possibly decrease the incidence of vascular complications associated with percutaneous intervention. This technique may allow for recanalization of totally occluded vein grafts with large clot burdens by using significantly less urokinase and shorter drug administration times than conventional infusion protocols. PMID- 9213025 TI - Time course of smooth muscle cell proliferation after local drug delivery of low molecular-weight heparin using a porous balloon catheter. AB - It has been reported previously that systemic application of low molecular weight heparin (LMWH) suppresses smooth muscle cell (SMC) proliferation after balloon angioplasty in experimental studies. However, the high concentration of heparin required for a beneficial effect may cause severe bleeding complications. The ideal situation to overcome the systemic side effects would be to administer LMWH locally and deep into the arterial wall, which became possible by the development of porous balloon catheters. The in vivo feasibility of local delivery of LMWH using the porous balloon has been assessed by delivering tritium-marked LMWH into rabbit carotid arteries. The efficacy of the system was investigated by using a second injury animal model. After development of an intimal plaque by electrical stimulation, 61 rabbits were treated with the porous balloon after balloon angioplasty. In 23 rabbits, local drug delivery was accomplished with a porous balloon catheter (35 holes, hole diameter 75 microns, 2.5 mm catheter diameter). LMWH was locally administered with 4 ml (solution 375 anti-Xa-units/ml) and 2 atm. To study the extent of restenosis and morphological changes, these animals were killed 3, 7, 14, 28, or 56 d after intervention. After staining (hematoxylin, van Gieson, BrdU, RAM 11, alpha-actin) procedures to quantify SMC proliferation, intimal macrophages and morphological analysis were performed. Porous balloon treatment led to an increase in intimal SMC proliferation rate in the early stage after intervention. However, during the following time period, a significant decrease of the proliferation rate as compared with the animals treated with balloon angioplasty alone could be observed, which resulted in an only moderate increase of the intimal layer after local drug delivery compared with balloon angioplasty alone. PMID- 9213026 TI - Local delivery of heparin to balloon angioplasty sites with a new angiotherapy catheter: pharmacokinetics and effect on platelet deposition in the porcine model. AB - The purpose of this study was to assess the efficacy of local heparin delivery to balloon angioplasty sites in an in vivo porcine model by using a newly designed angiotherapy catheter that allows for prolonged drug infusion while maintaining distal arterial perfusion. Protocols were designed to assess the safety of intracoronary drug delivery, the effect of infusion time and drug concentration on intramural heparin deposition, the distribution of heparin within the arterial wall, the histologic effects of local heparin delivery, the wash-out of intramurally deposited heparin, and the effect of heparin delivery on early platelet deposition following balloon injury in peripheral and coronary vessels. Local intracoronary delivery of heparin was well tolerated in all animals. Between 0.04 and 0.08% of infused heparin was intramurally deposited at the time of drug delivery, with longer infusion durations and higher concentrations of heparin resulting in greater intramural deposition. Autoradiography demonstrated homogenous distribution of heparin throughout the intima, media, and adventitia, with localization in the nuclei, cytoplasm, and extracellular space. Histologic analysis demonstrated no additional vessel trauma from local drug delivery beyond that seen with conventional angioplasty. Wash-out studies demonstrated a biexponential disappearance of intramurally deposited drug, with rapid release of heparin over the first 60 min and persistence of small amounts of drug for at least 7 d. Locally delivered heparin significantly attenuated the deposition of platelets in peripheral vessels, although a similar decrease in platelet deposition in the coronary arteries was not statistically significant. Local delivery of heparin directly to coronary angioplasty sites is possible with the use of a new angiotherapy catheter. Wash-out of heparin from the arterial wall is initially rapid, although drug is detectable for up to 1 wk following delivery. In porcine peripheral arteries, use of this technique significantly decreases early platelet deposition following balloon injury. PMID- 9213027 TI - Intramural drug delivery by direct injection within the arterial wall: first clinical experience with a novel intracoronary delivery-infiltrator system. AB - Local drug delivery at the lesion site in patients with coronary artery disease is being intensively studied to prevent restenosis after percutaneous coronary intervention. However, the effective penetration of the delivered agents into the vessel wall and delivery time remain considerable problems for all currently existing devices. A unique, new catheter has been invented, the infiltrator Angioplasty Balloon Catheter (IABC), which has the capability to allow intramural drug delivery by direct injection within the arterial wall. We describe the first clinical experience with this catheter. IABC is an angioplasty catheter with 3 lumens: one for inflating the balloon, one central for the guidewire, and a third for drug delivery. On the surface of the balloon there are 3 longitudinal strips of 6 injection needles, which on inflation stand 0.01" high, and are connected to the drug-delivery lumen. With inflation of the balloon, the needles penetrate the lesion, allowing drug delivery into the media of the vessel wall. We used the IABC in 17 patients (age = 58 +/- 9 years) undergoing coronary angioplasty. All patients were symptomatic, with significant lesions (13 LAD, 3 LCX, 1 RCA) and documented ischemia. Following initial dilatation with a conventional angioplasty balloon (stenosis from 72 +/- 8% to 26 +/- 14%, P < 0.001), the IABC was used to infiltrate the lesion with 0.4 ml (6,000 IU) of low-molecular-weight heparin (Fraxiparine). For the delivery, the IABC was inflated to 1-2 atm for 30-45 s, and the heparin was injected by hand in 5 s. Lesion residual stenosis and morphology remained unchanged after IABC use (26 +/- 14% to 22 +/- 11%, P = NS). In 10 patients, stent placement followed the IABC use. The decision to proceed with stent placement was made after the initial dilatation with the conventional balloon, and it was not influenced by the IABC use. Stent placement greatly improved the final result (for the whole patient group: 22 +/- 11% to 5 +/- 18%, for the stented patients: 22 +/- 13% to -7 +/- 10%, P < 0.001 for both). Hospital course was uneventful, with no electrocardiogram changes and normal cardiac enzymes for all patients. We have shown that the use of a unique new catheter (IABC) for intramural drug delivery in human patients undergoing coronary angioplasty is feasible and safe. This catheter is the first of a new generation of catheters and represents a significant step in local drug delivery. It is very promising as a vehicle to modify plaque behavior and potentially influence restenosis after angioplasty. PMID- 9213028 TI - Catheter-based local thrombolysis with urokinase: comparative efficacy of intraluminal clot lysis with conventional urokinase infusion techniques in an in vivo porcine thrombus model. AB - Local delivery of urokinase directly to the site of intraluminal clot using catheter-based technology has recently been introduced as a new technique to treat intracoronary thrombus and thrombus-containing stenoses. The purpose of this study was to compare the efficacy of urokinase therapy administered by local drug-delivery catheters with conventional urokinase-infusion techniques in dissolving intraluminal clot and intramurally depositing drug at the site of arterial injury in an in vivo porcine model. Five techniques of urokinase administration were studied in 65 pigs, including intravenous systemic bolus (1,000,000 units), guiding catheter infusion (500,000 units), local intraluminal infusion with a Roubin catheter (150,000 units), local infusion by the Dispatch catheter (150,000 units), and local delivery by the hydrogel-coated balloon (700 units). All five techniques were initially compared with respect to the quantity of intraluminal lysis of 123I-fibrinogen-labeled thrombus in an in vivo thrombus model. Conventional balloon angioplasty was also assessed in this model as a nonpharmacologic, mechanical control. In addition, all five techniques were compared with respect to the quantity and efficiency of intramural urokinase deposition at coronary angioplasty sites. In the in vivo thrombolysis experiments, the quantity of artificial clot lysis measured 6.8% for systemic therapy, 20.8% for guiding catheter infusion, 25.2% for Roubin catheter infusion, 62.8% for Dispatch catheter infusion, 98.8% for hydrogel balloon delivery, and 53.6% for conventional balloon angioplasty. Both the Dispatch catheter and the hydrogel balloon resulted in more clot lysis than the systemic, guiding catheter, or Roubin catheter approaches (P < 0.05). In comparison with conventional balloon angioplasty, only the hydrogel balloon resulted in higher levels of thrombus dissolution (P < 0.05). In the intramural deposition studies, the efficiency of urokinase delivery was 0.0004% for systemic therapy, 0.004% for guiding catheter infusion, 0.004% for Roubin catheter infusion, 0.08% for Dispatch catheter infusion, and 1.8% for hydrogel balloon delivery. The Dispatch catheter resulted in higher intramural drug levels than did all other techniques (P < 0.05), whereas the efficiency of urokinase deposition was higher with the hydrogel balloon than with all other approaches (P < 0.05). In the porcine model, it is subsequently concluded that local delivery of urokinase by catheter-based techniques can result in more complete lysis of intraluminal thrombus by using similar or lower doses of drug than by using conventional urokinase infusion techniques. Mechanical deformation of thrombus, possibly to increase the surface area available for thrombolysis and to physically disrupt clot, may be an important component of the mechanism of site-specific thrombolysis, particularly with the hydrogel balloon. Local delivery techniques also deposit significant quantities of urokinase at balloon angioplasty sites, creating an intramural reservoir of drug that may result in prolonged local thrombolysis. PMID- 9213030 TI - Local drug delivery with porous balloons in the rabbit: assessment of vascular injury for an improvement of application parameters. AB - OBJECTIVES: Sufficient intramural drug concentrations with the use of porous balloon catheters can be achieved with additional vascular trauma only. However, effective delivery of a potent drug even in deeper layers of the vessel wall might outweigh these traumatic side effects. Given the porous balloon catheter, the parameters of injection pressure and applied fluid volume will influence the interventional result. METHODS: We tested a 2.5-mm porous balloon (35 75-micron pores) in the right carotid artery of New Zealand rabbits and used injection pressures of 1, 2, and 5 atm and fluid volumes of 2 and 4 ml of low-molecular weight heparin solution in combination with the different parameters (n = 5 animals/group). In 50 rabbits, an intimal fibromuscular plaque was induced by using the electrostimulation model. Balloon dilatation and then application of the porous balloon was performed in 30 animals, 10 animals were only electrostimulated, and 10 animals served as a control group with balloon dilatation only. The vessels were excised 7 d after intervention, stained, and analyzed histomorpologically. Anti-Xa assays revealed the extent of systemically escaped drug, and serial cuts allowed for exact determination of vessel wall injuries. RESULTS: Effective local drug delivery could not be achieved with an injection pressure of less than 2 atm. Specific pressure-driven effects such as jet injuries could be identified. When the pressure was high enough for disruptive drug delivery (> or = 2 atm), fluid volumes of 4 ml led to loose elastic membranes and local thickening within the media. CONCLUSIONS: Sufficient intramural drug distribution using porous balloon catheters can be achieved with low injection pressures. Different fluid volumes strongly determine the extent of additional vascular injury. PMID- 9213029 TI - Efficacy of low-molecular-weight heparin delivery with the Dispatch catheter following balloon angioplasty in the rabbit iliac artery. AB - Local drug delivery can be achieved with active injection systems or passive contact of a compound with the arterial wall. The Dispatch catheter allows for passive diffusion of drugs from drug compartments while preserving blood flow through the central conduit. The aim of this study was to investigate whether a reduction in neointima formation can be achieved by local delivery of a limited amount of a highly concentrated solution of the low-molecular-weight heparin Reviparin. In 16 New Zealand white rabbits, successful balloon dilatation was performed in both iliac arteries, followed by local delivery of 4 ml Reviparin (1,000 IU/ml). The arteries were harvested at 7, 28, or 56 d following the procedure. The intimal cell layers increased substantially between 7 and 28 d following balloon dilatation with or without local drug delivery. The medial cell layers showed only a little increase. Proliferation of smooth muscle cells reached an early peak after 7 d, with a significantly higher proliferation index following local delivery. The maximum amount of macrophages in the intima and media was detected after 28 d. The lumen area decreased with time and was 0.6 +/- 0.7 mm2 in the local delivery group at 56 d compared with 0.5 +/- 0.5 mm2 in the control group. In conclusion, local delivery of Reviparin with the Dispatch catheter is safe and feasible. However, the infusion of highly concentrated low molecular-weight heparin over a short period of time did not result in a reduction of neointima formation and restenosis following balloon dilatation in the rabbit iliac artery. PMID- 9213031 TI - Localized delivery of heparin to angioplasty sites with iontophoresis. AB - Drug delivery by iontophoresis involves the application of an electric field to move selectively charged drug molecules across biological membranes. The purpose of this study was to assess the efficacy of intravascular iontophoresis in the local delivery of heparin to balloon angioplasty sites by using a recently designed iontophoretic catheter. In vivo heparin iontophoresis was assessed in 33 rats and 21 pigs in four protocols designed to measure the technical determinants of intramural drug deposition, the pharmacokinetics and localization of coronary delivery, and the effect of this technique on platelet deposition following balloon injury. First, iontophoresis of 3H-heparin into the aorta of 33 rats was performed to determine the effects of iontophoretic current, iontophoretic membrane balloon initiation pressure, iontophoresis time, and heparin concentration on intramural drug deposition. Second, iontophoresis of 3H-heparin was performed in 16 porcine coronary arteries to quantitate immediate drug delivery and subsequent wash-out over 24 h. Third, iontophoresis of fluorescent heparin was performed in 8 porcine coronary arteries to define intramural localization of locally delivered drug. Fourth, 111In-labeled platelet deposition was measured 1 h following balloon angioplasty and local iontophoretic heparin delivery in 16 porcine carotid and iliac vessels. Contralateral control vessels that were dilated with the same size balloon and treated with iontophoresis of saline served as controls. Rat aortic studies demonstrated that iontophoresis resulted in 13 times more intramural heparin deposition than passive delivery (passive: 0.3 +/- 0.4 microgram, iontophoresis: 4.6 +/- 1.6 micrograms, P < 0.0004). Iontophoretic membrane balloon inflation pressure had no significant effect on intramural drug deposition, but longer iontophoresis times and higher heparin concentrations resulted in higher levels of intramural heparin (P < 0.05). Porcine coronary studies demonstrated successful intramural deposition of heparin in all arteries without adverse electrical or hemodynamic sequelae, with persistence of the drug for at least 24 h. Localization studies demonstrated immediate deposition of fluorescent heparin in the intima and internal elastic lamina, with subsequent rapid diffusion of the drug into the media. Porcine platelet studies demonstrated that heparin iontophoresis decreased platelet deposition following balloon injury by approximately 66% compared with saline treated control vessels (heparin-treated: 1.46 +/- 2.51 x 10(8), control: 4.27 +/ 7.02 x 10(8), P = 0.001). This study has demonstrated that local intramural heparin delivery is feasible with an intravascular iontophoretic catheter. Following intracoronary heparin iontophoresis in the porcine model, intramural drug is detected for at least 24 h. Local delivery of heparin with this technique significantly decreases early platelet deposition following balloon injury in peripheral porcine arteries. PMID- 9213032 TI - Sustained local drug delivery to the arterial wall via biodegradable microspheres. AB - This study was designed to evaluate the feasibility of applying locally delivered polylactic acid microspheres for drug delivery to the arterial wall. To study drug persistence, rhodamine-loaded microspheres were infused into one carotid artery of 14 rabbits and plain rhodamine solution into the other by using a porous balloon. To study tissue response, plain microspheres and dexamethasone loaded microspheres were infused into the carotid arteries of another group of rabbits. To study the antiproliferative effects of locally delivered drug, 20 rabbits were subjected to overstretch balloon injury to both carotid arteries and divided into 4 groups: injury alone, plain microspheres, dexamethasone-loaded microspheres, and microspheres containing colchicine and dexamethasone. Fluorescent microspheres persisted in the vessel wall for 4 wk, whereas rhodamine without microspheres disappeared at 72 h. Histopathologic studies in arteries infused with unloaded microspheres showed inflammatory cell infiltrate with polymorphonuclear cells at 1 wk and macrophages and giant cells at 4 wk. Arteries infused with dexamethasone-loaded microspheres did not show any inflammatory cell infiltrate. Local delivery of dexamethasone or dexamethasone plus colchicine did not result in significant change in the intima-to-media ratio or in residual lumen following balloon injury. Polylactic acid microspheres may be used for prolonged delivery of drugs or other bioactive agents locally to the arterial wall. They induce an inflammatory reaction that is suppressable by dexamethasone in the microspheres. Dexamethasone or dexamethasone and colchicine delivered via this system, however, failed to reduce the degree of intimal hyperplasia after overstretch balloon injury to the rabbit carotid arteries. PMID- 9213033 TI - Infiltrator Angioplasty Balloon Catheter: a device for combined angioplasty and intramural site-specific treatment. AB - OBJECTIVES: We describe a new angioplasty device (Infiltrator Angioplasty Balloon Catheter; IABC) with intramural drug delivery capability. The conventional balloon part of the device, when inflated, dilates the vessel or has three rows of longitudinally mounted infiltrator nipples to penetrate the tunica media. Through an independent infiltrator port and nipples, drugs can be infiltrated directly into the vessel wall. METHODS: The device was tested in 117 normal coronary arteries of 58 farm pigs. RESULTS: (1) The infiltration procedure does not damage the vessel angiographically or histologically. At the infiltration site, the endothelial layer and the internal elastic lamina has a controlled interruption, and the infiltrated fluid is distributed among the medial layers, causing a mild focal edema and medial thickening (1.8 times on average). (2) Rhodamine tracer is circularly and evenly distributed through the whole width of the vessel wall within 10 min after infiltration. (3) Two weeks after the infiltration procedure, the medial layer reveals mild local thickening and remodelling without luminal narrowing. (4) Of the intramurally infiltrated Tcm99 labeled surfurcolloid, 83.8% is detectable at 5 min after the delivery procedure by gamma camera. (5) The IABC, if oversized, is able to achieve angiographic dilatation in normal pig coronary arteries, causing histologic damage similar to or less than that induced by conventional balloon dilatation. CONCLUSIONS: The IABC can deliver fluid-phase substances directly into the vessel wall with microliter accuracy and with 90% efficiency without significant acute and subacute damage to the vessel wall. It is also suitable for combined dilatation and local drug delivery. PMID- 9213034 TI - Site-specific intravascular administration of drugs: history of a method applicable in humans. AB - Demonstration and quantification of site-specific intracoronary administration of pharmacological compounds has been limited thus far to animal experimental models. Recently, a method applicable in humans has been developed. The aim of this study is to give an overview on the available methods to visualize and quantify intravascularly administered "labeled" drugs in animals and to describe the historical development of a method now applied in the clinical arena. The potential of this approach is briefly summarized. PMID- 9213036 TI - Pharmacokinetics and tissue localization of antisense oligonucleotides in balloon injured pig coronary arteries after local delivery with an iontophoretic balloon catheter. AB - When delivered locally to the arterial wall by passive fluid transfer systems such as perforated balloons, water-soluble compounds in aqueous solution are not readily taken up by tissue, show low levels of cellular localization, and are quickly lost by wash-out. One approach to improve delivery is addition of an "active" component to the catheter system to change the nature of the drug-to tissue interaction. Using an iontophoretic balloon catheter to deliver antisense oligonucleotide (ODN) to pig coronary arteries after balloon angioplasty, we determined the quantity and localization of ODN in the tissue. By radiolabeling, 7.3 +/- 2.4 micrograms ODN was present at 30 min, 1.5 +/- 0.6 at 2 h, 0.52 +/- 0.35 at 24 h, and 0.26 +/- 0.11 at 7 d. By fluorescent labeling, circumferential medial uptake and adventitial delivery at the site of medial injury was observed, with primarily cellular localization. The iontophoretic catheter thus appears to be a useful device for ODN delivery to arterial tissue. PMID- 9213035 TI - Endoluminal local delivery of PCNA/cdc2 antisense oligonucleotides by porous balloon catheter does not affect neointima formation or vessel size in the pig coronary artery model of postangioplasty restenosis. AB - Localized delivery of antisense oligonucleotides directed against cell cycle regulatory proteins has been proposed as a means to prevent restenosis after angioplasty. To test whether single endoluminal delivery of a combination of proliferating cell nuclear antigen (PCNA) and cell-division cycle 2 kinase (cdc2) antisense might affect restenosis, we delivered 2 ml of lipid-complexed PCNA/cdc2 antisense oligomers (1.35 mg) to the coronary arteries of pigs after balloon overstretch angioplasty (AS group) and performed planimetric histomorphometry on arterial sections of the tissue, harvested at 4 wk. Compared with controls receiving 3'-5' reversed sequence oligomers (REV group), there were no differences in absolute intimal area (AS 1.36 +/- 0.08 mm2, REV 1.23 +/- 0.10 mm2, P = NS), intimal area normalized to extent of injury (AS 0.67 +/- 0.03, REV 0.77 +/- 0.10, P = NS), or vessel perimeter (AS 7.72 +/- 0.19 mm, REV 7.36 +/- 0.22 mm, P = NS). We conclude that single endoluminal delivery of antisense against key cell cycle regulatory proteins does not affect neointima formation or vessel size in this model of restenosis. PMID- 9213037 TI - Tomographical anatomy of the pelvis, pelvic floor, and related structures. AB - The sectional anatomy of the pelvic floor was studied in plastinated sections of adult pelves by computed tomography and by magnetic resonance imaging. In sectional anatomy, the levator ani is composed of three portions that can be clearly distinguished by their planes of cleavage and by the course of their fiber bundles. No muscular connections are found between the levator ani portions and the pelvic organs. The fascia of the levator ani in always interposed between the muscle and the pelvic organs. The sectional anatomy of the sphincter ani externus reveals a subdivision into a subcutaneous and a deep portion. Although the puborectalis portion of the levator ani and the deep portion of the sphincter ani externus are more or less continuous, in sectional anatomy they can be distinguished due to their different origins and attachments. PMID- 9213038 TI - Celiacomesenteric trunk. AB - The celiac trunk is the widest ventral branch of the abdominal aorta. The unusual embryological development of the ventral splanchnic arteries can lead to considerable variations. During the dissection of a 54-year-old male cadaver as a rare variation, a celiacomesenteric trunk was observed. The rare occurrence of this variation is stated to be 1%-2.7%. As in the other case, the celiac trunk and the superior mesenteric arteries arose from a common trunk at the level of L1. This case of celiacomesenteric trunk is described in detail, which can be of value in the operative procedures of the upper abdomen. PMID- 9213039 TI - Anatomical study of the synovial plicae of the hip joint. AB - Observations and measurements of the synovial plicae of the hip joints were made on 63 embalmed cadavers. The cadavers were divided equally among three age groups (fetuses, children, and adults). Our observations showed that the plicae appeared in two forms (flat and villous) and were mainly confined to the external surface of the lower medial part of the acetabular labrum (labral plicae), the base of the ligament of the head of the femur (ligamental plicae), and along the reflecting line of the synovial membrane on the base of the femoral neck (neck plicae). The ligamental plicae were well padded with a fibroelastic pad of fat filling the acetabular fossa, and the neck plicae were far away from the articular surfaces of the joint; as a result, neither was likely to be injured or entrapped during joint movements. The labral plicae were larger than the ligamental or neck plicae and had an incidence of 73.8% in the fetal group. The fetal plicae were found only after the fetal age of 5 months. In nine cases of the child and adult groups, the labral plicae extended between the articular surface of the femoral head and the lower part of the acetabulum during medial rotation of the thigh. When the thigh was rotated laterally, the plicae in six of the same cases could be returned to their original positions. In the remaining three cases, there was continual impingement. PMID- 9213040 TI - Anorectal abscesses: need for accurate anatomical localization of the disease. AB - Anorectal abscesses constitute a common problem of the perianal area. Considerable morbidity is expected if an immediate and anatomically correct drainage procedure is not performed in a timely manner. Acute infection of an anal gland leads to the formation of the anorectal abscess, while the chronic stage of the infection appears as a fistula in ano. It is imperative to perform operations with accurate anatomical knowledge; this is particularly true in the case of fistula-in-ano and anorectal abscesses, for which inappropriate surgery can lead to disastrous results. Here we report our experience with anorectal abscesses in various locations in 14 patients. Four of the 14 had a transrectal drainage. The internal (transrectal) or external drainage of the anorectal abscesses depends mainly on the mechanism of their formation and the anatomical relationship of the abscess to the levator ani. Apart from a death occurring 1 month after drainage due to a cause not related to suppurative disease (subdural hematoma), all patients had an uneventful recovery and were discharged from hospital after a mean stay of 1.2 days. A brief and practical description of the macroscopic anatomy of the area will assist in understanding better the selection of the appropriate route of drainage of anorectal abscesses. PMID- 9213041 TI - Unreported anatomical variation of septum pellucidum. AB - During gross anatomy dissections of the brain, a developmental abnormality of the septum pellucidum was found in a 31-year-old male cadaver. Other parts of the central nervous system in this cadaver were normal in every aspect. Histological samples were taken from the neighboring areas of this abnormality, and they were examined under light microscope and scanning electron microscope. In this abnormality of the septum pellucidum, the two laminae of the septum pellucidum were fused together and there was a hole located 1 cm anterior to its apex. The maximum diameter of the hole was 0.5 cm in the sagittal plane and 0.6 cm in the vertical plane. In the light microscopic and scanning electron microscopic examinations, the free margin of this foramen was regular, and the surrounding tissue was intact and histologically unique to the septum pellucidum. Ependymal cells were present at the free margin of the foramen. Cavum vergae, cavum septum pellucidum, and agenesis of the septum pellucidum are described in the literature. These three abnormalities are seen in cadavers usually with histories of schizophrenia and other psychiatric or neurologic disorders. In a retrospective study, the cadaver with this abnormality had a history of schizophrenia and no history or signs of any kind of brain or head operation. As far as we could ascertain, the abnormality described here has not been reported previously. PMID- 9213042 TI - Scalene muscles and the brachial plexus: anatomical variations and their clinical significance. AB - Anatomical variations may be clinically significant, but many are inadequately described or quantified. Variations in neck anatomy are important to surgeons performing surgical procedures in this region. Thirty-two female and 19 male adult cadavers were studied. The commonly described anatomical relationship of the brachial plexus (BP) lying between the anterior scalene (AS) and middle scalene (MS) muscles was found in only 60% of instances. Scalenus minimus was present in 46% of instances (bilateral in 14 cadavers). The most common variation was the penetration of the AS by the C5 and/or C6 ventral rami. The C5 and C6 roots may fuse before piercing AS (15% cases, bilateral in 4 cadavers), or the C5 root alone pierce the belly of AS (13% cases, bilateral in 3 cadavers). The roots also may pierce AS independently (6% cases, bilateral in 1 cadaver). In 3%, the C5 root was found to be completely anterior to AS. PMID- 9213043 TI - Rare case of high origin of the ulnar artery from the brachial artery. AB - Arterial variations in the arm are numerous and occur at the level of the axillary, brachial, radial, and ulnar arteries as well as in the palmar arches. We report on a high branching site of the ulnar artery. A high branching brachial artery was found in a 72-year-old white female during a dissection course. The brachial region was then dissected carefully and the preparation steps were documented. The axillary artery, after entering the arm, was located posterior to the junction of the two roots of the median nerve, just 2 cm distal to the latter, and divided into the ulnar and the radial arteries. The radial artery was located medial to the median nerve in the arm and approached the lateral side of the arm to reach the cubital fossa. Just distal to its origin, the ulnar artery ran laterally crossing ventral to the median nerve, thereafter supplying the biceps brachii muscle with three branches from a common stem. The ulnar artery then approached the medial side of the arm, crossed ventral to the median nerve again and took its course toward the cubital fossa as usual. This high bifurcation of the brachial artery and the abnormal course of the ulnar artery is of interest to clinicians; in particular vascular and plastic surgeons and radiologists. PMID- 9213044 TI - Variation in the triceps brachii muscle: a fourth muscular head. AB - Routine cadaver dissection has resulted in the identification of a fourth head of the triceps brachii muscle on the left side in one specimen. This novel arrangement demonstrated a single tendon arising from the proximal posteromedial aspect of the humeral shaft, distal to the shoulder capsule. The tendon of this fourth head passed along the medial aspect of the humerus and gave way to a muscle belly on the medial surface of the distal one-third of the humerus. The tendon of the fourth head passed directly over the neurovascular bundle containing the radial nerve and deep brachial artery at approximately the point where the neurovascular bundle entered the radial sulcus. This close positional relationship between the tendon of the fourth head, the radial nerve, and the deep brachial artery has prompted us to speculate on the possible clinical significance of this finding in relation to radial nerve palsy and arterial compression. Additionally, the position of the muscle belly, lying in close proximity to the ulnar groove, invites speculation on the role of the fourth head in cases of snapping elbow. To the authors' knowledge, a description of the muscular fourth head of the triceps as seen in the present work has not been noted in previous literature. PMID- 9213045 TI - Survey of the educational roles of the faculty of anatomy departments. AB - The anatomy dissection laboratory is a unique experience where medical students begin the transition from layman to physician, and may be a student's first experience with death. Attitudes developed there may influence interactions with future patients and their families. Consequently, anatomy faculty are in a position to recognize emotional issues that students may confront and to guide them toward becoming humane physicians. We surveyed anatomy faculty to assess acceptance of this expanded role and their means of meeting these obligations. A spokesperson for the anatomy department at each US and Canadian medical or osteopathic college (n = 142) was surveyed. One hundred three (73%) questionnaires were returned. Respondents overwhelmingly (93% agree or strongly agree) accept an educational role that includes helping students to become caring physicians and dealing with death and dying. Seventy-nine percent agree or strongly agree that the anatomy laboratory can affect students later relationships with patients. Time for laboratory orientation is limited (55%, 1 hour or less) and is used to address technical topics, such as rules for student behavior. Most departments (58%) have four or more memorial activities to acknowledge the contribution of the donors. The anatomy faculty who responded to the survey accept responsibility for acculturating preclinical students to medicine. Respondents identified additional orientation topics and expanded memorial activities to accomplish this goal. PMID- 9213046 TI - Attitudes and reactions of Arab medical students to the dissecting room. AB - A questionnaire on the emotional and psychological reactions of Arab medical students to the dissecting room (DR) was distributed to 272 students in four successive pre-clinical and clinical years in the same academic year (1993-1994) at Sultan Qaboos University (SQU) Medical College; 205 students responded. Varying degrees of fear on first entering the DR was reported by 46%. The most frequent reactions were recurring visual images of cadavers (total 38%) and temporary loss of appetite (total 22.5%). Students' reactions were most commonly elicited by the smell of the DR (total 91%) and by fear of infection (total 62%). The most frequent method of coping with such fears was by rationalization (total 65%). Significant gender differences (P < 0.05) were found concerning all aspects of the DR experience. Female students showed higher levels of fear, reported stronger physical and behavioral reactions, were more disturbed by certain stimuli in the DR, and used certain coping methods more frequently than their male peers. The need for appropriate psychological preparation of students before studying human cadaveric anatomy is discussed. PMID- 9213047 TI - Problem in diagnostic imaging. AB - The staging of malignant disease for metastatic spread requires a sound understanding of the normal anatomy and possible anatomical variants as well as the expected pathology. This article illustrates this in the case of a young man with seminoma of the testis undergoing CT imaging to assess for para-aortic metastatic spread. A potential pitfall that may mislead the unwary is demonstrated. PMID- 9213048 TI - Nerve supply of the breast with special reference to the nipple and areola: Sir Astley Cooper revisited. AB - Cooper in 1840 described mammary branches from the 2nd-6th intercostal nerves, and noticed that the nipple was supplied by branches which lay close to the surface of the gland. Eckhard (1850) divided the mammary branches into superficial branches to the skin and nipple, and deep branches to the glandular tissue and nipple, but many later authors ignored those findings. After the second World War, cosmetic surgery of the breast made further research critical, as surgeons strove to design operations which would retain its shape and preserve postoperative sensation. Craig and Sykes (1970) described mainly anterior branches from the 3rd, 4th and 5th intercostal nerves passing through the glandular tissue of the breast and along the line of the ducts to the nipple, while Farina et al. (1980) concluded that the nipple was supplied solely by superficial lateral branches of the 4th nerve. Using improvements in dissecting technique learned from microsurgery, Sarhadi et al. (1996) found that the nipple was innervated by the lateral cutaneous branch of the 4th intercostal nerve, by two branches, one passing superficial to the gland, and the other through the retromammary space, and by variable lateral and medial additional branches from the 2nd-5th nerves. These branches came to lie superficially and formed a subdermal plexus under the areola. This account is uncannily close to Cooper's original description; it is a reassuring, if sobering, conclusion that his early account remains one of the most reliable. PMID- 9213050 TI - Familial spastic paraplegia: evidence for a fourth locus. AB - Autosomal dominant familial spastic paraparesis (AD-FSP) is a genetically heterogeneous disorder of the central nervous system characterized by a progressive spasticity of the legs. One gene causing AD-FSP (FSP1) has recently been mapped to chromosome 14q, another gene (FSP2) to chromosome 2p, and a third gene (FSP3) to chromosome 15q. We now report a large Dutch family with AD-FSP without linkage to any of these chromosomes, providing evidence for a fourth locus (FSP4). PMID- 9213051 TI - The use of and familiarity with neurological eponyms at the close of the 20th century by Dutch neurologists. An inventory. AB - In order to get an impression on the opinion of the use of neurological eponyms we sent questionnaires on 205 eponyms to 850 members of the Netherlands Association for Neurology. The responses by 256 (30%) were analyzed. A positive correlation was found between age and the knowledge of eponyms. The best known eponyms belong to category II (tests and manoeuvres). Surprisingly, many of the responding colleagues did not prefer descriptive terms to eponyms: 57% of the eponyms were preferred by more than 50% of the respondents. PMID- 9213049 TI - Relation of neuropsychological and magnetic resonance findings in amyotrophic lateral sclerosis: evidence for subgroups. AB - The relationship between neuropsychological impairments and changes in cranial MR images was investigated in a group of 74 consecutive patients with the sporadic form of amyotrophic lateral sclerosis (ALS). Neuropsychological tests included measures of frontal lobe function, memory, intelligence, and attention. Compared with a control group, a significant impairment of the ALS group emerged for the areas of visual attention, inhibition of response alternatives, visual memory, and word generation. These neuropsychological impairments did not show a relation to clinical status of the patients. Likewise, MR parameters derived by computer assisted planimetric analysis showed a ventricular enlargement and parenchymal atrophy in the ALS group compared with age-matched controls. When ALS patients were assigned to two subgroups differing on the basis of the neuropsychological tests by cluster-analysis the cluster with the significant impairment also showed a pronounced change for the MR-parameters while the subgroup showed essentially normal neuropsychological performance. This pattern suggests that subgroups of ALS with differential impairment of neuropsychological functions can be defined. PMID- 9213052 TI - Cerebellar ataxia, dementia, pyramidal signs, cortical cataract of the posterior pole and a raised IgG index in a patient with a sporadic form of olivopontocerebellar atrophy. AB - Middle-aged patients who initially present with a progressive cerebellar ataxia, in the absence of a known familial pattern are often referred to under the descriptive diagnosis of 'idiopathic' late onset cerebellar ataxia. If these patients in time develop additional pyramidal or extrapyramidal features then they should be labeled as olivopontocerebellar atrophy (sOPCA). This case report describes a patient with OPCA with cerebellar ataxia as the presenting and most prominent feature in combination with dementia, pyramidal signs, cortical cataract of the posterior pole and a raised IgG index in cerebrospinal fluid. To the best of our knowledge this combination of signs and symptoms have not been described before. PMID- 9213053 TI - Amnesia following infarction in the right retrosplenial region. AB - We present a case of retrosplenial amnesia following an infarction in the right retrosplenial region. A 62-year-old male showed mild weakness of the left hand, dizziness and gait disturbance. He also noticed that he could not perceive objects that he saw as real, but could perceive an object as real only by touching it. Magnetic resonance imaging (MRI) showed an infarction in the splenium of the corpus callosum and retrosplenial region on the right side. There was no aphasia or apraxia, but mild topographic disturbance was present. Intelligence was normal, but amnesia was noted. Both verbal and visual memory were disturbed equally. This case suggests that memory plays a role in the right retrosplenial region. PMID- 9213054 TI - Swallowing difficulty in Parkinson's disease. AB - Dysphagia is a frequent and potentially serious complication of Parkinson's disease (PD). We examined the oropharyngeal swallowing ability in 19 PD patients (15 men and 4 women, mean age 68.42 years, mean Hoehn and Yahr stage 1.8) using modified barium swallow before and after administering oral levodopa (in combination with benserazide). Twelve (63.2%) patients demonstrated objective evidence of swallowing abnormalities; although only six patients (31.6%) had subjective complaints. Vallecula sinus and pyriform sinus residues were the most frequent abnormalities (47.4% and 42.1%); followed by delayed swallowing reflex (26.3%). Three patients demonstrated silent aspiration. In the 12 patients with abnormal swallowing, six (50%) showed objective improvement after levodopa treatment, while the remaining six showed no change. Of the former group of six, one patient showed improvement in the oral phase, but deterioration in the pharyngeal phase. We concluded that PD patients had a high percentage of objective swallowing abnormalities which could be reduced in half of the patients through the administration of levodopa treatment. PMID- 9213055 TI - Collins' law. Prediction of recurrence or cure in childhood medulloblastoma? AB - Collins' law, known as period of risk for recurrence (PRR), has been discussed as a reliable scheme for the prediction of recurrence or cure of embryonal tumours including medulloblastomas. The PRR is defined as the age at diagnosis plus 9 months of gestation. According to Collins' law, a patient who has no clinical evidence of recurrence within this time period is considered cured. We have applied this system to 66 patients (age range from 0 to 47 years) who underwent surgery for medulloblastoma at a single center between 1975 and 1990. Three patients in whom a recurrence could not be verified died within 6 months. Eight patients (12.1%, age range from 1 to 14 years) survived the PRR without relapse and are free of risk for recurrence according to Collins' law. Of the remaining 55 patients, 35 showed a relapse within the PRR with a mean latency of 18 months (0-78); Twenty patients are alive with no recurrence as yet, but are still within the PRR Three patients had non-recurrent benign brain tumours not connected to the initial tumours with a mean latency of 10 (8-14) years after medulloblastoma surgery. Thus, all our patients fulfilled the criteria for Collins' law. In conclusion, a patient's risk for medulloblastoma recurrence is significantly higher before than after the completion of the PRR. Therefore, the PRR is a helpful prognostic parameter that provides additional information about the risk of medulloblastoma recurrence. A medulloblastoma cure cannot be presumed with certainty by Collins' law because relapses after the PRR have been reported in other studies (1.4% of the cases). Intracranial lesions detected after a long follow-up might be radiotherapeutically induced second tumours. PMID- 9213056 TI - Management of glioblastoma multiforme: with special reference to recurrence. AB - Between 1985 and 1995, 46 patients underwent craniotomy for glioblastoma multiforme. The mean age was 47, varying from 9 to 71 years. The influence of such prognostic factors as age, preoperative Karnofsky score, extent of resection, tumour site, tumour size, radiotherapy, reoperation as well as initial symptoms upon survival were studied. Of these, gross complete removal, radiotherapy, preoperative Karnofsky score, and reoperation were shown to be statistically significant to the survival time according to logrank and univariate tests. However, age, preoperative Karnofsky score, tumour size and temporal localisation remained as significant factors in multivariate analysis. The overall median survival was 53 weeks, with no patients surviving more than 3 years. Of the patients, 41% survived over a year and 8.6% lived over two years. Twenty-six patients developed a recurrent mass after an interval of 32 weeks. The median interval time from operation to recurrence was longer in those patients who underwent gross removal than in those who had a subtotal resection, 28.2 against 20 weeks (P < 0.05). Of patients who had a recurrent mass, 16 were reoperated on, with a subsequent median survival time of 26.5 weeks. Our experience suggests that the survival of patients with glioblastoma depends on many factors, including radical surgery as an initial step. In addition, the gross total removal of the tumour also delays the development of recurrence. PMID- 9213057 TI - Inter-hemispheric scissure, a rare location for a traumatic subdural hematoma, case report and review of the literature. AB - Subdural interhemispheric hematomas (ISH), though not really rare, are quite an uncommon complication of head traumas. This condition is more frequent in childhood, where it is generally considered as a part of a more complex syndrome, called 'Shaken Children Syndrome', usually pointing out child abuse. Although a head injury is very often considered the cause (in about 80-90% of the cases), possible predisposing factors such as coagulopathies, alcohol abuse or anticoagulant therapy can also be considered. Furthermore, as the rupture of an intracranial aneurysm has also occasionally pinpointed as a possible cause, this event should be kept in mind in order to be able to address exactly both the diagnostic and therapeutical procedures. A new conservatively managed case is thereby described. A review of the literature with particular attention drawn to the diagnosis and the different therapeutical possibilities is also elaborated. PMID- 9213058 TI - Cerebellar mutism: report of two unusual cases and review of the literature. AB - Mutism is not a common condition following cerebellar damage. Mutism following posterior cranial fossa surgery was first reported by Rekate et al. and Yonemasu in 1985. Since then, many case reports of mutism have appeared in the English literature. Very few cases developed mutism following brain stem surgery. Although mutism has been described in patients with head injury, only one case of mutism caused by a cerebellar injury has been reported, to our knowledge. We report on two patients in which the cerebellar mutism following a radical excision of an exophytic brain stem glioma and cerebellar injury developed. We reviewed the relevant literature and discussed the mechanism of cerebellar mutism. PMID- 9213059 TI - Isolated dural metastasis from colon cancer. AB - We report a case of isolated dural metastasis from colon cancer. Neuroradiologic and intraoperative findings indicated that the tumor was extra-axial, and histopathologic examination resulted in the correct diagnosis. Notable neuroradiologic findings of the present dural-based tumor were: diploic fat MRI signal disappeared in the tumor associated calvaria where bony change was absent on bone-window CT; abnormal subgaleal and epidural tissues were seen on MRI, both of which contained clusters of adenocarcinoma cells histopathologically. These findings suggest an infiltrative, malignant character, and help to differentiate between benign and malignant dural-based tumors. PMID- 9213060 TI - Lumbosacral plexus neuropathy: a case report and review of the literature. AB - Lumbosacral plexus neuropathy (LSPN) is an idiopathic clinical syndrome characterized by the sudden onset of neuropathic pain, followed by weakness and sometimes sensory disturbances in the distribution of the lumbosacral plexus. Prognosis is usually favourable, although complete recovery may take several months to years. LSPN is the lumbosacral counterpart of the neuralgic amyotrophy syndrome (idiopathic brachial plexus neuropathy). We present a patient who initially was misdiagnosed with a radicular syndrome, but illustrates the typical signs and symptoms of LSPN. We also give clinical and electromyographical criteria for the diagnosis of LSPN and review the literature. PMID- 9213061 TI - Familial dyslexia associated with cavum vergae. AB - Three members of one family, diagnosed as dyslexic, are described. All of them have variations of midline cavity: cavum vergae or cavum septum pellucidum, diagnosed by neuroradiological examination. In contrast, the non dyslexic members of the same family have no neuroanatomical congenital variations. We raise the possibility of a functional correlation between the dyslexia and the anatomical findings in the affected members of this family. PMID- 9213062 TI - Borjeson-Forssman-Lehmann syndrome: two severely handicapped females in a family. AB - We report on a case of Borjeson-Forssman-Lehmann syndrome (BFS) in which two sisters appear with the clinical picture of severe mental handicap, dysmorphic features and grand mal seizures. Their mother presents the above symptomatology in a milder degree. It is one of the few reports in literature where females develop BFS in its typical clinical form. PMID- 9213063 TI - The value of MRI in a case of Tolosa-Hunt syndrome. AB - We report a case of Tolosa-Hunt syndrome (THS) in which the lesion has been demonstrated by magnetic resonance imaging (MRI), computed tomography (CT), and angiography. A healthy 23-year-old man developed an acute painful ophthalmoplegia on the right side. CT and MRI scans revealed asymmetric enlargement of the right cavernous sinus with contrast enhancement extending down to the region of trigeminal ganglion. MRI further delineated the detailed anatomical structures of the region and excluded any infiltration of the surrounding tissues by a mass lesion. Cerebral angiography showed a significant decrease in the calibration of petrous segment and a mild decrease in the calibration of cavernous segment of the ipsilateral internal carotid artery. The patient was treated with oral prednisone, 100 mg daily. Neurological findings totally subsided after 2 weeks on corticosteroid and MRI showed resolution of the lesion in the cavernous sinus. The patient was symptom-free for 6 months after discharge. Our findings have suggested that MRI is the most valuable imaging technique for demonstration and follow-up of lesions in the cavernous sinus that are directly responsible for the symptoms of THS and the lesions can be more extensive than was currently believed. PMID- 9213064 TI - The cerebellum in Alzheimer's disease. AB - The cerebellum is a relatively neglected area of the Alzheimer's disease (AD) brain, probably because it was formerly thought to be spared by the disease. However, a number of pathological changes have now been revealed in the AD cerebellum, principally by immunocytochemical studies, including widespread deposits of diffuse amyloid, ubiquitin-immunoreactive dystrophic neurites, and increased microglia, but tau-immunoreactive neurofibrillary tangles have not been seen. Although the observed changes may be merely epiphenomenal to the pathological processes occurring in the AD neocortex and hippocampus, the morphological and immunocytochemical differences between AD cerebral cortex and cerebellar cortex may nonetheless give insights into the molecular factors involved in the development of the neuropathological lesions of AD brain. PMID- 9213065 TI - Procedural memory in patients with mild Alzheimer's disease. AB - Motor, perceptual, and cognitive skill learning abilities of mild Alzheimer's disease (AD) patients were compared to sex-, age-, and education-matched controls. We excluded patients who were unable to perform each skill learning task with a predetermined criterion. In those who completed the task, skill learning was as good as in normal controls. On the cognitive and perceptual skill learnings, some of the AD patients, whose cognitive but not declarative memory functions were more severely impaired than in those who completed the whole session, failed to complete the task, while all patients could complete the motor task. These results support that view that patients with mild AD can acquire motor, perceptual, and cognitive skills and that the neural system subserving procedural skill is not related to the neural systems for declarative memory. PMID- 9213066 TI - Progressive formation of neuritic plaques and neurofibrillary tangles is exponentially related to age and neuronal size. A morphometric study of three geographically distinct series of aging people. AB - Neuronal size and the incidence of neuritic plaques (NPs) and neurofibrillary tangles (NFTs) were morphometrically and quantitatively studied in the entorhinal cortex of 300 autopsied individuals without dementia in three geographically distinct series (Brazil, Germany and Japan), and an additional series including 30 clinically diagnosed Alzheimer's disease patients. The mean ages at onset of NPs and NFTs were similar between the three normal series, and the incidence of NPs and NFTs increased exponentially with age, but at different rates. A correlation was found between larger neuronal size and higher incidence of NPs and NFTs. Neuronal size distribution largely seemed to account for the differences between the series. While the onset of neurodegeneration may be tightly programmed, i.e., in a species-specific manner, our data support the idea that the incidence of NPs and NFTs and the progression from NPs to NFTs may vary remarkably, depending on neuronal size. PMID- 9213067 TI - The disturbances of object vision and spatial vision in Alzheimer's disease. AB - Thirty-one patients with mild-to-moderate Alzheimer's disease (AD) underwent a test battery of complex visual tasks. We assessed the scores using a principal factor analysis to elucidate the underlying deficits. There were three independent factors: The first factor included the tasks of identifying and comparing forms of visual stimuli. The second factor consisted of digit span and digit symbol tasks, and the third factor consisted of a specified visual counting task. We considered these three factors as representing the dysfunctions of object recognition, general attention and spatial recognition, respectively. These results underline the disturbances of the two visual systems, object vision and spatial vision, in early-AD patients. PMID- 9213068 TI - Somatostatin and neuropeptide Y in cerebrospinal fluid: correlations with severity of disease and clinical signs in Alzheimer's disease and frontotemporal dementia. AB - Alzheimer's disease (AD) and frontotemporal dementia (FTD) are the most common types of progressive neurodegenerative disorder in our catchment area. The distribution of cortical degeneration in FTD is mainly the reverse of that in AD, while there are both differences and similarities in the clinical characteristics. Somatostatin and neuropeptide Y (NPY) are neuropeptides with a widespread distribution in the human cerebral cortex. Somatostatin is involved in the regulation of hormone release from the anterior pituitary and may act as a neurotransmitter-modulator. NPY is a potent anxiolytic neuropeptide. Somatostatin and NPY coexist in the cerebral cortex, basal ganglia and in amygdaloid complexes. The present study of AD (n = 34) and FTD (n = 22) analyses the cerebrospinal-fluid (CSF) levels of somatostatin-like immunoreactivity and NPY like immunoreactivity and correlates their levels to 54 different clinical items, such as restlessness, anxiety, irritability and depression. The CSF levels of the two neuropeptides somatostatin and NPY were significantly correlated in FTD (p < 0.02), but not in AD. Several significant correlations to the clinical signs were found: in AD disorientation and dyspraxia, and in FTD agitation, irritability and restlessness. Somatostatin showed a significant negative correlation with severity of dementia in AD (p < 0.013). PMID- 9213069 TI - Apolipoprotein-E genotyping in Alzheimer's disease and frontotemporal dementia. AB - Alzheimer's disease (AD) and frontotemporal dementia (FTD) are characterized by progressive neuronal loss and microvacuolization, although with different distributions of cortical involvement. In contrast to AD there is no amyloid, senile plaques or tangles in FTD. The involvement of chromosome 19 in AD has been associated with apoliprotein E (ApoE) and the epsilon 4 gene frequency has been related to increased risk and early onset of AD. Our analysis of frequency of the ApoE alleles in 38 patients with AD, 21 patients with FTD and 29 normal controls indicates an association of both AD and FTD with an increased frequency of the epsilon 4 allele and in AD also with homozygosity for epsilon 4. Our results might indicate that ApoE epsilon 4 is an important aggravating and pathoplastic factor in the presence of genetic and other determinants for the development of AD or FTD. A significantly higher epsilon 2 frequency in our FTD material compared to AD and normals might also indicate a connection with the distribution of cortical degeneration. PMID- 9213070 TI - Distinguishing between patients with depression or very mild Alzheimer's disease using the Delayed-Word-Recall test. AB - The present study investigated the accuracy of an extended version of the Delayed Word Recall (DWR) test in distinguishing patients with very mild Alzheimer's disease (AD) (Mini Mental State Examination score > or = 23) from community dwelling depressed/dysthymic patients. The DWR test was administered to 26 non depressed patients who, at the time of DWR administration or on follow-up, fulfilled NINCDS/ADRDA criteria for probable AD, and to 20 age-matched non dementing patients with a diagnosis of major depression (n = 12) or dysthymia (n = 8) according to DSM-III-R criteria. Sensitivity and specificity were, respectively, 96 and 100% for DWR free recall, and 92 and 100% for DWR recognition. In this study both DWR free recall and recognition measures were highly sensitive and specific in distinguishing very-mild-AD patients from depressed/dysthymic patients. The investigation of more severely depressed patients is warranted. PMID- 9213071 TI - Staging of Alzheimer-type pathology: an interrater-intrarater study. AB - The staging method proposed by Braak and Braak is based on the sequential accumulation of neurofibrillary pathology in the cerebral cortex. Unlike the currently used diagnostic criteria for Alzheimer's disease (AD) it does not take into consideration the age of the patients and whether they were demented or not for the establishment of the pathological stage of the disease. To examine the interobserver reliability of the method we performed an inter- and intrarater study using the Braak staging method in 41 brains. The agreement between the examiners and between the diagnoses of the same examiner at different times was almost perfect, the kappa statistics reaching values above 0.90. These findings indicate that the staging, relying on the differential distribution of neuritic pathology in the brain in AD, is a reliable and reproducible method for the description of AD-related pathology. This makes it suitable for brain-banking and research purposes. PMID- 9213072 TI - The vascular dementia of Fabry's disease. AB - Fabry's disease, a rare X-linked disorder of glycosphingolipid metabolism, can present as an insidious dementia in middle or later life. This genetic disorder produces a deficiency of alpha-galactosidase A which results in the deposition of glycosphingolipids in blood vessel walls in the brain as well as in the kidney, heart, peripheral nerves, and other organs. Among the cerebrovascular manifestations of this disorder is a vascular dementia from involvement of multiple small penetrating blood vessels. Fabry's disease is a consideration in the workup of an otherwise unexplained vascular dementia, particularly in males less than 65 years of age. PMID- 9213073 TI - Senile dementia and apolipoprotein E4. PMID- 9213074 TI - Pharmacokinetic and pharmacodynamic studies of (R)-8-hydroxy-2-(di-n propylamino)tetralin in the rat. AB - Racemic 8-OH-DPAT, (R,S)-8-hydroxy-2-(di-n-propylamino)tetralin, has become the prototype 5-HT1A receptor agonist. The enantiomers of 8-OH-DPAT have similar affinities to the 5-HT1A receptor, but the (R)-enantiomer is a full agonist, whereas the (S)-enantiomer is a partial agonist. This communication describes the dose- and time-response relationships of behavioural (5-HT behavioural syndrome, cage-leaving response), physiological (body temperature) and biochemical (5-HT turnover, 5-hydroxytryptophan accumulation) effects of (R)-8-OH-DPAT in rats. A high-performance liquid chromatography (HPLC)-UV method for determination of plasma and brain concentrations of (R)-8-OH-DPAT was developed, permitting studies of the pharmacokinetics of the drug. The concentrations of 8-OH-DPAT in brain were several fold higher than in plasma, and there were large variations in (R)-8-OH-DPAT concentrations between brain regions (highest in the hippocampus). (R)-8-OH-DPAT peaked in plasma at 5 min and in brain at 15 min after subcutaneous administration. The 5-HT1A behavioural syndrome peaked within 5 min after administration and disappeared after 30 min, when brain concentrations were still high. The hypothermic and biochemical responses developed gradually and were maximal at 45-60 min post injection, when both plasma and brain concentrations were declining. Thus, there was not a simple relationship between the kinetics and the dynamics of (R)-8-OH-DPAT. These results prompt further studies on the pharmacokinetics of 8-OH-DPAT within the central nervous system. PMID- 9213075 TI - Altered protein phosphorylation in the rat brain following chronic lithium and carbamazepine treatments. AB - Lithium and carbamazepine (CBZ) alter levels of specific kinase-activating second messengers generated by adenylate cyclases and the phosphoinositide system. Thus, lithium and CBZ may change endogenous protein phosphorylation mediated by cyclic AMP-dependent protein kinase (PKA) and protein kinase C (PKC). The present study aimed at comparing the chronic effects of lithium and CBZ on protein phosphorylation in the rat brain by using quantitative autoradiography. Long-term treatments yielded plasma levels within the therapeutic range. In the particulate hippocampal fraction PKA-mediated phosphorylation of a 42 kDa protein and PKC mediated phosphorylation of a 88 kDa protein were decreased after lithium treatment. In the cortical particulate fraction approximately 30% reduction in the PKA-mediated protein phosphorylation of several proteins was observed after lithium and CBZ treatments. In the same fraction, CBZ treatment significantly reduced PKC-mediated phosphorylation of several substrates by 30-40%. PKA activity was significantly reduced in cortex, but not in the hippocampus. Thus, both drugs exhibited fraction and region specificities. PMID- 9213076 TI - The comparative effects of haloperidol, (-)-sulpiride, and SCH23390 on c-fos and c-jun mRNA expressions, and AP-1 DNA binding activity. AB - The c-fos and c-jun mRNA expressions were measured by the RNase protection assay method following intraperitoneal injection of haloperidol, (D1 and D2 receptor antagonists), (-)-sulpiride, (D2 receptor antagonist), and SCH23390, (D1 receptor antagonist). Haloperidol and (-)-sulpiride increased their mRNA expressions in a dose-dependent manner, peaking at 30 min after injection followed by a gradual decline. The SCH23390 did not induce expression of either c-fos or c-jun mRNA. A significant decrease of c-fos as well as c-jun mRNA expression was found due to pretreatment with SCH23390 (1 mg/kg i.p.) followed by injection of (-)-sulpiride (20 mg/kg i.p.). The results suggest that the expression of these mRNAs is closely related to the dopamine D2-like antagonism. Administration of haloperidol or (-)-sulpiride increased AP-1 DNA binding activity with similar manner of dose dependence, whereas their activities were reduced by Fos and Jun antibodies, implying that AP-1 components, transcriptional factors, forming due to Fos and Jun were actually activated by either haloperidol or (-)-sulpiride. PMID- 9213077 TI - In vivo cyclic AMP responses in rat brain are not modified by chronic electroconvulsive shock. AB - In vivo microdialysis was used to determine the effects of chronic electroconvulsive shock (ECS), given daily for 10 days, on the cyclic AMP (cAMP) responses to noradrenaline (NA) and to forskolin in the cortex of freely-moving conscious rats. Microdialysis probes were inserted on the final day of ECS administration, and cAMP responses measured the following day. Chronic ECS did not modify the responses to either NA or forskolin. These results differ markedly from those previously obtained with incubated slices of rat brain. It is concluded that care is needed in interpreting results obtained by either ex vivo or in vitro methods. PMID- 9213078 TI - PET neuroimaging with [11C]venlafaxine: serotonin uptake inhibition, biodistribution and binding in living pig brain. AB - The brain binding kinetics and distribution of the antidepressant venlafaxine, labelled with 11C in the O-methyl position, was studied by PET after intravenous injection in anesthetized pigs. In addition, venlafaxine's action on serotonin (5 HT) uptake was studied in vitro in blood platelets obtain from humans or pigs. Venlafaxine resembled imipramine, paroxetine and citalopram in causing a dose dependent inhibition of 5-HT uptake in blood platelets from pigs and humans. Venlafaxine-derived radioactivity entered the living brain readily and showed higher binding potentials in diencephalic and telencephalic regions than in cerebellum. Acute administration of an antidepressant drug (i.e. imipramine, citalopram or paroxetine) enhanced the distribution and altered the binding of venlafaxine in certain brain regions. The findings show that [11C]venlafaxine is not an ideal PET radiotracer mainly because of its relatively low binding potentials and its lack of specificity for the 5-HT transporter in living brain. PMID- 9213079 TI - A naturalistic study of paroxetine in premenstrual syndrome: efficacy and side effects during ten cycles of treatment. AB - Eighteen women with severe premenstrual syndrome (PMS) (premenstrual dysphoric disorder, PMDD) were treated openly with paroxetine for 10 consecutive menstrual cycles. Dosage was flexible (5-30 mg/day); also, the patients were free to chose between continuous medication and medication in the luteal phase only. The rating of premenstrual irritability, depressed mood, increase in appetite, and anxiety/tension was markedly lower during treatment with paroxetine than before, and this reduction in symptomatology appeared unabated for the entire treatment period. Sedation, dry mouth, and nausea were common side-effects but declined during the course of the trial; in contrast, reduced libido and anorgasmia, which were reported by almost 50% of the participants, were not improved with time. The results indicate that the beneficial effects as well as the sexual side-effects of serotonin reuptake inhibitors persist unchanged for at least 10 consecutive cycles of treatment. PMID- 9213080 TI - Tricyclic antidepressant-induced extrapyramidal side effects. AB - Two cases of tricyclic antidepressant-related extrapyramidal side effects are reported and, the authors review the literature describing these effects. Despite clear case reports, these side effects are not well known. Given the wide prescription of tricyclic antidepressants (TCA) and the low number of case reports, the prevalence of these side effects is indeed low, but clinical implications exist. The extrapyramidal symptoms induced by TCA alone are acute or tardive dyskinesia, akathisia, myoclonus, rabbit syndrome and dystonia. These symptoms seem to be non age-related, but often dose-related, and were responders to antiparkinsonian agents or propranolol. The factors that predispose an individual to the development of these side effects are not completely understood. Some risk factors such as prior exposure to neuroleptics and/or lithium or estrogens could facilitate the development of these side effects. In some cases, they can disappear even though the same dose of TCA is continued, and they do not seem to be a drug class reaction. The susceptibility of each individual patient to the development of these disorders may be limited to only one or a few of these agents. PMID- 9213081 TI - Side effects from increased doses of carbamazepine on neuropsychological and posturographic parameters of humans. AB - Patients taking anticonvulsant drugs display a broad spectrum of side-effects. Particularly, in the beginning of treatment and with increasing doses of carbamazepine, side effects such as dizziness, ataxia, drowsiness and reduction of alertness occur, which improve some days after the dose has reached a stable level. Our aim was to find objective parameters for grading these side effects and to differentiate between neurophysiological and neuropsychological side effects of carbamazepine in a clinical situation. Twenty-two patients with trigeminal neuralgia were included for a follow-up study with increasing carbamazepine doses (0 mg to 600 mg). The effect of carbamazepine on postural stability was quantified by posturography. Different neuropsychological tests to study cognitive effects of carbamazepine were performed. The composite equilibrium score showed a significant reduction of postural stability with increasing doses of carbamazepine. In sensory analysis the somatosensory ratio was significantly influenced by increased doses of carbamazepine during the study. Mean reaction time of tonic alertness and physical alertness varied significantly with different doses of carbamazepine. There was a significant influence in patients attention during trail making tests and divided attention tests with increase in carbamazepine. In conclusion our observations show that the rate of change of carbamazepine doses is an important determinant of cognitive and motor functions in the phase of increasing doses. PMID- 9213082 TI - The effects of sulpiride on psychomotor performance and subjective tolerance. AB - In many European countries, the substituted benzamide sulpiride is used with antidepressant indication in the dosage range of 150-300 mg on an outpatient population. This raises the concern of possible impairments of psychomotor performance in this dosage range. To address this question, the psychometric effects of 300 mg of sulpiride in comparison with placebo in 12 healthy volunteers was evaluated in this study. In a randomised, double-blind, two-way, within-subjects (cross-over) design, visuomotor performance was assessed using time estimation, critical flicker fusion, and choice reaction time tasks at baseline and 4 h after oral administration of either 300 mg of sulpiride or placebo. In addition, self-ratings on subjective well-being were obtained. Results were evaluated using analysis of covariance (ANCOVA) with baseline levels as covariates. In healthy subjects, 300 mg of sulpiride caused no alteration in time estimation and choice reaction movement time, whereas critical flicker fusion frequency was lower and choice-reaction decision time were prolonged under medication. Self-rating scales showed no significant differences between sulpiride and placebo. Subjects were not able to tell whether they received placebo or sulpiride. This study indicates that sulpiride is subjectively well tolerated at a dosage of 300 mg. However, using psychometric methods, effects are demonstrable that can be interpreted as a reduction of excitatory arousal without causing the subjective experience of sedation. These results call for caution when prescribing the drug to outpatients. PMID- 9213083 TI - Pathophysiological significance of cerebral perfusion abnormalities in major depression-trait or state marker? AB - Considerable data support the existence of impaired regional cerebral blood flow in major depression. However, it is unclear whether the impairment in brain function in major depression is a "state" marker, reversible upon remission, or an enduring trait phenomenon. We have studied brain technetium-99m hexamethylpropyleneamineoxime (Tc-99m HMPAO) uptake ratios in healthy subjects of various ages, in depressed patients before and after electroconvulsive therapy (ECT) and in healthy subjects before and after administration of fluoxetine. Analysis of our findings, presented along with research data of other groups, strongly suggests that reduced cerebral perfusion in major depression is reversible by successful treatment. Furthermore, since fluoxetine had little effect on cerebral perfusion in healthy subjects, and ECT had little effect on cerebral perfusion in depressed patients who did not respond to treatment, we contend that increases in perfusion represent remission rather than treatment effect. Therefore, reduced perfusion in major depression appears to be a state marker and not a trait abnormality. PMID- 9213084 TI - Effects of clomipramine on plasma amino acids and serotonergic parameters in panic disorder and depression. AB - We examined the effects of long-term (six months) treatment with the serotonin potentiating tricyclic antidepressant clomipramine on several serotonergic parameters in panic disorder and depressive patients. Serotonin (5-HT) levels in blood, platelets and plasma were significantly reduced to 4%, 3% and 28% of their respective baseline values. In addition, the plasma level of tryptophan was also significantly reduced, although the decrease was only 16%. Three months after discontinuation of clomipramine treatment, 5-HT in blood and platelets reached baseline values again, while the plasma 5-HT level was still reduced to 68% of pretreatment values. Unexpectedly, the plasma tryptophan concentration was even lower at this time-point than after six months of treatment. These results show that clomipramine not only has an effect on 5-HT levels in blood, platelets and plasma, but also on plasma tryptophan concentration. We speculate that low plasma tryptophan after treatment may constitute a risk for the recurrence of psychopathology. PMID- 9213085 TI - Successful treatment of salbutamol-induced panic disorder with citalopram. PMID- 9213086 TI - Functional adaptations in patients with ACL-deficient knees. PMID- 9213087 TI - Interactions between muscle glycogen and blood glucose during exercise. AB - Muscle glycogen and blood glucose are important substrates for contracting skeletal muscle during exercise. The possibility exists for considerable interaction between muscle glycogen and blood glucose and their effects on muscle glucose uptake and glycogenolysis, respectively. Increases in blood glucose availability have little effect on net muscle glycogen utilization during prolonged, strenuous cycling exercise, but this may be dependent upon the mode and intensity of exercise. No data exist on the direct effect of reduced blood glucose levels on muscle glycogen metabolism. In rats, studies using the perfused hindlimb suggest an inverse relationship between muscle glycogen levels and glucose uptake. A similar conclusion can be drawn from a number of human studies albeit, on occasion, from indirect evidence. The influence of muscle glycogen on glucose uptake involves effects on both membrane glucose transport and intracellular glucose metabolism. Such a relationship would, under conditions of adequate muscle glycogen availability, limit muscle glucose uptake, thereby preserving the relatively small blood glucose supply for the brain, nerves, and blood cells, which are dependent on it for their energy metabolism. PMID- 9213088 TI - Thermoregulation at rest and during exercise in healthy older adults. AB - Epidemiological evidence of increased mortality among older men and women resulting from hyper- and hypothermia should not be interpreted as implying that aging per se confers an intolerance to environmental extremes. Relatively few studies have attempted to delineate the effects of chronological age from concomitant factors (such as decreases in VO2max, lowered habitual activity levels, alterations in body composition, etc.) in determining thermoregulatory responses to rest and exercise in hot environments. When the effects of chronic diseases and sedentary life-style are kept to a minimum, heat tolerance appears to be minimally compromised by age. VO2max is more important than age in predicting body temperature during exertion, even though some of the physiological mechanisms associated with heat dissipation (especially control of skin blood flow and distribution of cardiac output) are closely associated with chronological age. Dehydration effects may be magnified in older individuals, and rehydration may be compromised by age-related differences in thirst sensitivity and renal function. The efficacy of various interventions in improving thermoregulatory responses of older individuals (e.g., aerobic training and heat acclimation) has not been studied adequately. Older men and women are capable of acclimating to hot conditions, but the time course of physiological changes underlying acclimation with age may be different. Another intervention that holds promise is hormone replacement therapy in postmenopausal women, which acutely affects temperature regulation and control of body fluids in a positive direction. The chronic effects of hormone replacement therapy on thermoregulation during exercise and environmental stresses are not known. Tolerance to cold exposure under resting conditions may be less dependent on age and aerobic fitness than on body composition. Studies of older and younger subjects exercising in cold environments are lacking altogether, including important studies into the possible preventive effects of regular physical activity on physiological responses to cold stress. PMID- 9213089 TI - Effects of endurance exercise on adipose tissue metabolism. PMID- 9213090 TI - The cellular stress response to exercise: role of stress proteins. PMID- 9213091 TI - High-protein diets, "damaged hearts," and rowing men: antecedents of modern sports medicine and exercise science, 1867-1928. AB - There are many criteria by which advances in "sports medicine" and "exercise science" may be assessed. Historical developments in the basic and applied sciences--and in clinical practices, as well--may best be explored by those who are experts in particular fields. It is encouraging that more studies of this type have begun to appear. Studies of the evolution of organizations that have fostered various aspects of physical training, exercise science, and sports medicine may provide another useful approach. Whereas 910 individuals participated in the 1976 ACSM Annual Meeting, in 1992, attendance reached 3661. In 1954 (the year the American College of Sports Medicine was organized), Index Medicus listed approximately 250 items under the headings Athletics, Exercise, and Physical Education. In 1993, some 4000 citations were reported as being about Athletics, Exercise, Physical Fitness, Exercise-Related Physical Therapy, and Sports Medicine. According to the Congress Proceedings of the Third IOC World Congress on Sport Sciences (September 16-22, 1995), 225 individuals made presentations. More than 1650 women and men representing a vast array of contributing fields participated in symposia, lectures, and poster sessions during "Physical Activity, Sport, and Health"--The 1996 International Pre-Olympic Scientific Congress (July 10th-14th). What advances await in the 21st century? PMID- 9213092 TI - Bone, exercise, and stress fractures. PMID- 9213093 TI - Physical activity among persons with disabilities--a public health perspective. AB - Regular physical activity, sports participation, and active recreation are essential behaviors for the prevention of disease, promotion of health, and maintenance of functional independence. This health behavior is essential for persons with and without disabilities. Population-based surveys have consistently demonstrated that persons with disabilities are less likely to be physically active, compared to persons without such limitations. However, these observations are based on relatively few surveys and are dependent on physical activity assessment methods that may not be sensitive and specific enough for persons with disabilities. Studies clearly demonstrate that many persons, representing a variety of selected disabilities, can adapt to increased levels of physical activity, as evidenced by alterations in various components of physical fitness. More importantly, other studies consistently provide evidence that participation in regular physical activity among persons with selected impairments and disabilities results in improved functional status and quality of life. Further efforts are critically needed in the area of the development of physical activity assessment methodology for persons with disabilities. Methods need to be developed that will provide survey researchers and those in public health the capacity to measure and monitor activity patterns of persons with disabilities. This information is important not only for public health officials but also health policy analysts, service providers, and disability advocacy groups. Further understanding of the role of physical activity in the maintenance of function and independence among persons with disabilities is needed. The understanding of environmental and social barriers to physical activity among persons with disabilities needs further exploration. Finally, physical activity determinants research among persons with disabilities, including the role of assistive technology as well as maximizing the intrinsic capacity of functional anatomy and physiology, needs to be addressed. PMID- 9213094 TI - Exercise testing and training in patients with malignant arrhythmias. AB - Patients with malignant arrhythmias are a challenge when they present to the laboratory for exercise testing and training. The complex metabolic and electrophysiological changes that occur during and after exercise may suppress or potentiate arrhythmias and alter the efficacy and safety of antiarrhythmic agents. The presence of an ICD adds a level of complexity to the study; however, a general understanding of how ICDs function and a review of the programmed parameters before the study will result in a lesser likelihood of complication and more efficient and appropriate action, should a complication occur. Exercise testing and training are valuable methods for the evaluation and treatment of patients with malignant ventricular arrhythmias. Exercise testing is a useful adjunctive technique in the exposure of arrhythmia and the evaluation of the efficacy and proarrhythmic potential of antiarrhythmic agents. These patients are excellent candidates for rehabilitation and exercise training programs, which may result in increased functional capacity and improved quality of life. With this knowledge and adherence to the appropriate guidelines for patient selection and testing, exercise testing and training of patients with malignant arrhythmias can be prescribed and conducted in a safe, controlled manner in an atmosphere of confidence and professionalism. PMID- 9213095 TI - Visceral obesity, insulin resistance, and dyslipidemia: contribution of endurance exercise training to the treatment of the plurimetabolic syndrome. PMID- 9213096 TI - The regulation of gene expression in hypertrophying skeletal muscle. PMID- 9213097 TI - Force transmission in skeletal muscle: from actomyosin to external tendons. AB - The actual path of force transmission in skeletal muscle from actomyosin interaction to tension at the tendinous insertion site is poorly understood. Within the muscle cell, endo- and exosarcomeric cytoskeletal proteins create series and parallel connections between contractile proteins resulting in a meshwork across which force can be transmitted in practically any direction with respect to the fiber axis. At the surface membrane, connections between the intermediate filament system, dystrophin, and specialized membrane complexes provide the route of force transmission to the extracellular matrix material. Finally, parallel and series connections between muscle fibers allow radial and longitudinal forces to converge on the connective tissue matrix. This complex pathway will certainly be the subject of future studies in muscle biology, biomechanics, and physiology. PMID- 9213098 TI - Baroreflex regulation of blood pressure during dynamic exercise. AB - From the work of Potts et al. Papelier et al. and Shi et al. it is readily apparent that the arterial (aortic and carotid) baroreflexes are reset to function at the prevailing ABP of exercise. The blood pressure of exercise is the result of the hemodynamic (cardiac output and TPR) responses, which appear to be regulated by two redundant neural control systems, "Central Command" and the "exercise pressor reflex". Central Command is a feed-forward neural control system that operates in parallel with the neural regulation of the locomotor system and appears to establish the hemodynamic response to exercise. Within the central nervous system it appears that the HLR may be the operational site for Central Command. Specific neural sites within the HLR have been demonstrated in animals to be active during exercise. With the advent of positron emission tomography (PET) and single-photon emission computed tomography (SPECT), the anatomical areas of the human brain related to Central Command are being mapped. It also appears that the Nucleus Tractus Solitarius and the ventrolateral medulla may serve as an integrating site as they receive neural information from the working muscles via the group III/IV muscle afferents as well as from higher brain centers. This anatomical site within the CNS is now the focus of many investigations in which arterial baroreflex function, Central Command and the "exercise pressor reflex" appear to demonstrate inhibitory or facilitatory interaction. The concept of whether Central Command is the prime mover in the resetting of the arterial baroreceptors to function at the exercising ABP or whether the resetting is an integration of the "exercise pressor reflex" information with that of Central Command is now under intense investigation. However, it would be justified to conclude, from the data of Bevegard and Shepherd, Dicarlo and Bishop, Potts et al., and Papelier et al. that the act of exercise results in the resetting of the arterial baroreflex. In addition, if, as we have proposed, the cardiopulmonary baroreceptors primarily monitors and reflexly regulates cardiac filling volume, it would seem from the data of Mack et al. and Potts et al. that the cardiopulmonary baroreceptor is also reset at the beginning of exercise. Therefore, investigations of the neural mechanisms of regulation involving Central Command and cardiopulmonary afferents, similar to those being undertaken for the arterial baroreflex, need to be established. PMID- 9213099 TI - Physical activity, adolescence, and health: an epidemiological perspective. PMID- 9213100 TI - Perception of physical exertion: methods, mediators, and applications. PMID- 9213101 TI - Laryngeal cancer: quality-of-life and cost-effectiveness. AB - BACKGROUND: The need to account for the cost of care is being imposed on clinicians by the corporate purchasers of health care. The cost-effectiveness of treatment for laryngeal cancer should be based on measurable and relevant clinical outcomes. Quality-of-life (QL) is a very important outcome, especially if survival is similar between two treatments. METHODS: Using QL data gathered prospectively from a group of 46 patients with glottic cancer, the effects of radiotherapy (RT) are compared with those of surgery (total laryngectomy). RESULTS: Surgery results in greater measurable dysfunction, especially of speech, but with respect to psychological functioning and general well-being, there are no major QL differences between patients treated surgically and those receiving RT. CONCLUSIONS: Cost-effectiveness of surgery, from the perspective of patient based QL outcome, is very similar to that for RT in advanced cancer of the larynx. Each center needs to carefully consider its outcomes and overall average costs per patient before deciding which treatment regimen is to be preferred. These results have implications for the organ-sparing regimens in the treatment of advanced laryngeal cancer. PMID- 9213102 TI - Nasal lymphoma: results of local radiotherapy with or without chemotherapy. AB - BACKGROUND: Primary nasal lymphoma is a rare disease. Although most patients are initially seen in early Ann Arbor stages, their prognosis is poor. The prognostic significance of local tumor bulk has not been well studied. METHODS: Twenty-one patients with nasal lymphoma treated between 1985 and 1992 were retrospectively studied. Sixteen patients (76%) below the age of 75 years received combined radiotherapy and chemotherapy. One young patient with early disease and 4 elderly patients had radiotherapy alone. Twelve cases (57%) were diagnosed as pleomorphic T-cell lymphoma based on typical histologic features alone. Immunophenotyping was performed in 10 cases; 8 were T cell and 2 were B cell. Seventeen patients (81%) had Ann Arbor clinical stage IE disease, and 4 had stage IIE disease. The local tumor extent was assessed by endoscopy in all patients and by computerized tomography (CT) in 14 patients. Eleven local tumors (52%) extended to the posterior ethmoids, sphenoid sinus, orbit, or beyond. Using a T-stage system, the prognostic significance of local tumor bulk was evaluated for stage IE patients. RESULTS: At a median follow-up time of 16.8 months, the lymphoma recurred in 13 patients; 10 patients had systemic relapse and 10 patients, local relapse. The 5 year actuarial overall survival rate was 24%. Complete response to chemotherapy was achieved in 5 of 16 patients (31%). Four of the 6 patients who remained alive and disease-free were chemotherapy complete responders. Among stage IE patients, those with early and those with advanced local disease did not have significantly different survival. CONCLUSIONS: In view of the high systemic and local relapse rates, more-effective chemotherapy is needed to improve the survival rates, and the role of combined chemotherapy and radiotherapy should be evaluated. Further studies are required to identify patients at high risk of relapse for clinical trials with investigational treatment. PMID- 9213103 TI - Expanded application of selective neck dissection with regard to nodal status. AB - BACKGROUND: The efficacy of extending the application of selective neck dissection to include more-extensive neck disease in patients with squamous carcinoma of the upper aerodigestive tract remains controversial. METHODS: A review of all patients undergoing selective neck dissection at a single institution during a 5-year period was undertaken. The analysis was conducted on 82 patients who received 94 selective neck dissections as part of initial therapy for management of squamous carcinoma of the upper aerodigestive tract, including: oral cavity, oropharynx, larynx, and hypopharynx. RESULTS: Forty-six of the 94 dissected necks were supraomohyoid dissections, and 48 were lateral neck dissections. Sixty-five percent of patients were followed a minimum of 2 years and formed the cohort for final analysis. There were eight regional recurrences, three of which occurred in the contralateral, undissected neck. The regional recurrence rate for all patients undergoing selective neck dissection, with or without radiotherapy, according to pathologic N status was as follows: NO (1/33), 3%; N1 (1/8), 12.5%; and multiple positive nodes (3/26), 11.5%. A comparison of recurrence rates with respect to extent of neck disease (N0-N1 versus multiple positive nodes) for both types of neck dissection did not demonstrate significant differences; supraomohyoid neck dissection, p < .5; lateral neck dissection, p < .25. CONCLUSIONS: There exists an expanded role for selective neck dissection in selected patients with primary squamous cell carcinoma of the upper aerodigestive tract and multiple N+ cervical disease. The selection of patients who are candidates for selective lymphadenectomy should be based on pathoanatomic considerations with reference to the primary site of tumor and demonstrated level(s) of metastatic involvement. PMID- 9213104 TI - Evaluation of cervical nodal necrosis in nasopharyngeal carcinoma by computed tomography: incidence and prognostic significance. AB - PURPOSE: The purpose was to study the prognostic value of contrast-enhanced computed tomography (CT) nodal necrosis in nasopharyngeal carcinoma. PATIENTS AND METHODS: One hundred sixty-one patients with newly diagnosed nasopharyngeal carcinoma and nodal metastases were reviewed. Forty patients also received cisplatin-based neoadjuvant chemotherapy in addition to radiotherapy. Nodal necrosis was defined as presence of hypodense areas in more than 33% of the node. Nodal response rate to chemotherapy, overall nodal control rate, local control rate, distant failure rate, overall relapse-free survival rate, and overall and cause-specific survival rates were compared between patients with and without nodal necrosis. Multivariate analysis was also performed. RESULTS: The incidence of nodal necrosis was 22.9%. Overall nodal response rates to chemotherapy were 88.9% (8/9) in patients with nodal necrosis and 74.2% (23/31) in those without. No significant differences in nodal control rate, local control rate, distant failure rate, and overall and cause-specific survival rates were found. Five-year overall relapse-free survival rate was lower in patients with cervical nodal necrosis (36%) as compared with those without (53%, p = .04). Multivariate analysis, however, did not confirm cervical nodal necrosis to be an independent prognostic factor. CONCLUSIONS: Presence of nodal necrosis in nasopharyngeal carcinoma does not affect nodal response to chemotherapy and nodal control by radiotherapy with or without chemotherapy. Cervical nodal necrosis does not appear to be an independent factor in predicting treatment outcome. Further studies to correlate nodal density with oxygenation status as well as tumor cell kinetics are warranted. PMID- 9213106 TI - Quantification of surgical margin shrinkage in the oral cavity. AB - BACKGROUND: Obtaining adequate surgical margins, free of tumor, is crucial for success in oncologic surgery. The head and neck surgeon often finds that the tumor-free margin reported from histopathologic measurement is significantly smaller than the margin measured in-situ. It was the purpose of this study to quantify the change in size of mucosal and muscle surgical margins following excision, formalin fixation, and slide preparation of tongue and labiobuccal tissue in a canine model. METHODS: Ten mongrel dogs under general anesthesia for a concurrent project were used in this study. Changes in mucosal and muscle dimensions around custom-made brass disks, one with a needle depth gauge, were measured immediately following excision after formalin fixation and after slide preparation. RESULTS: The mean shrinkage from initial resection to final microscopic assessment of the lingual surface mucosal margins was 30.7% (p < 0.0001). The deep tongue margin shrank 34.5% (p < 0.0001). The mean shrinkage of the labiobuccal mucosal margin was 47.3% (p < 0.0001). In all cases, the greatest proportion of shrinkage occurred immediately upon resection. CONCLUSIONS: From the in-situ measurement by the surgeon to final pathologic evaluation on the microscope slide, the measured dimensions of oral cavity mucosal and tongue muscle margins shrink significantly. To obtain 5 mm of pathologically clear margin an in-situ margin of resection of at least 8 to 10 mm needs to be taken. Studies reporting clinical correlation of recurrence and survival information with surgical margin status should include a detailed description of the technique used to determine the reported surgical margin status. PMID- 9213105 TI - Tumor angiogenesis as a prognostic indicator in T2-T4 oral cavity squamous cell carcinoma: a clinical-pathologic correlation. AB - BACKGROUND: Tumor angiogenesis has been shown to correlate with tumor size, metastatic potential, and prognosis in breast and other cancers. Studies in head and neck cancer have suggested a similar correlation, but results have been inconclusive. This study was performed to determine the correlation between angiogenesis and oral tumor behavior. METHODS: Tumor angiogenesis was evaluated in 31 T2-T4 primary oral cavity squamous cell carcinomas by quantitating the microvessel density with two different anti-endothelial cell antigens, factor VIII antigen (FVIIIAg) and CD-31. The stains were compared to assess whether these antigens yielded complimentary results. The microvessel densities were correlated with T stage and N stage and patient survival. RESULTS: FVIIIAg and CD 31 staining yielded consistent microvessel densities, but FVIIIAg was generally more uniform and easier to interpret. Increasing microvessel density was seen with increasing T stage and N stage; however, there was considerable overlap and no correlation with survival. CONCLUSIONS: These results suggest that oral tumors are less angiogenesis dependent than tumors in other sites. Tumor angiogenesis, as currently measured, is not of value in predicting tumor aggressiveness in patients with oral cavity carcinoma. PMID- 9213107 TI - A comparison of masticatory function in patients with or without reconstruction of the mandible. AB - BACKGROUND: The functional benefits of mandibular reconstruction following a composite resection remain unclear. Although microvascular surgical techniques have dramatically increased the predictability of bone and soft-tissue reconstruction towards presurgical anatomic norms, the specific factors responsible for improved function remain controversial. Objective measures of masticatory function need to be more clearly determined before the predictability and efficacy of reconstructive approaches is established. METHODS: We evaluated objective measures of oral function and patient reports of function in 10 reconstructed mandibulectomy patients, 10 without reconstruction, and 10 controls. Measures of oral function included bite force assessed at the first molar and incisal edge, a measure of tongue and cheek function, and patient reports of food they could eat. RESULTS: Both reconstructed and nonreconstructed patients presented decreased biting force, a more restricted diet, and compromised cheek and tongue function as compared with normals. However, reconstructed patients had significantly better measures of tongue function and ability to eat a varied diet than did nonreconstructed patients. Of the objective measures used to measure masticatory performance, bite force was poorly correlated, whereas measures of tongue function strongly correlated with successful mastication. CONCLUSION: Both reconstructed and nonreconstructed patients presented with a significant functional deficit when compared with normals, with reconstructed patients having better overall function than nonreconstructed patients. PMID- 9213108 TI - The longus colli muscle flap for reconstruction of the lateral pharyngeal wall. AB - BACKGROUND: Full-thickness lateral pharyngeal wall (LPW) defects are difficult to reconstruct, whether the larynx is preserved or removed (extended total laryngectomy). A simple, reliable reconstructive method using local tissue which optimizes wound healing and functional results would allow partial laryngectomy more often, without incurring the cost, donor site morbidity, and increased operative length of regional or free flaps. My objective was to propose use of the longus colli muscle as a reconstructive flap for defects of the LPW. METHODS: Results of using the longus colli muscle flap (LCMF) in a series of 16 patients with primary tumors of the pharyngeal wall or pyriform sinus are presented. The majority had surgery and planned postoperative radiotherapy. RESULTS: There were no wound infections or fistulas. One of 2 previously radiated patients had a transient wound-healing problem. Although 88% of the patients were stage III and IV and 50% had T3-4 primary tumors, there were only 2 local failures, for a local control rate of 88%. Corresponding cancer-free survival was 69% (median follow-up of 22 months). Two thirds of the patients took all or some food by mouth, and of the 12 with larynx preserved, 58% were decannulated, and 11 had a good to normal voice. CONCLUSIONS: The reliability of wound healing and absence of negative impact on oncologic and functional results validate use of the LCMF as a reconstructive option for defects in the LPW at both the oropharynx and hypopharynx levels. PMID- 9213109 TI - Maxillectomy and its classification. AB - BACKGROUND: Many adjectives are used to describe maxillectomy procedures, such as radical, total, extended, subtotal, medial, partial, and limited. The variety of nomenclature in our own Service database testifies that much confusion exists. METHODS: We have reviewed a 10-year experience with 403 maxillectomies performed between 1984 and 1993. Based on our retrospective reassessment, the operations were grouped into one of three categories. The term "limited" (LM) was applied to any maxillectomy which primarily removed one wall of the antrum. Designated "subtotal" (SM) was any procedure which removed at least two walls, including the palate. We listed as "total" (TM) only those who had a complete resection of the maxilla. Hospital charts were selectively reviewed, and each of the three types of maxillectomy was analyzed to determine the histology and site of the index cancers and the incidence of complex reconstruction. RESULTS: We determined that the maxillectomy performed in 230 patients (57%) was a LM. Tumor site and extent defined five different approaches in this cohort: peroral, 73; medial maxillectomy, 53; anterior craniofacial, 43; upper cheek flap, 42; and transfacial, 19. Subtotal maxillectomy or TM was performed in 135 and 38 (34% and 9%, respectively), almost 90% of whom had a cheek flap approach. Only 51 patients had an orbital exenteration, including 27 of the 38 (71%) of those who had a TM. Complex repair was employed in a total of 63 patients (16%), most often in those having TM (14 of 38, 37%). CONCLUSIONS: Classification of maxillectomy either as LM, SM, or TM is useful and feasible. To define a LM, the portion of the maxilla removed (ie, palate, anterior wall, medial wall) must be specified. For any maxillectomy, the access used should be listed, and the surgeon should indicate whether the maxillectomy has been extended to include adjacent structures. PMID- 9213110 TI - Hyperbaric oxygen therapy for wound complications after surgery in the irradiated head and neck: a review of the literature and a report of 15 consecutive patients. AB - BACKGROUND: Radiotherapy, which is often used for cancer in the head and neck, leads to damage of tissue cells and vasculature. Surgery in such tissues has an increased complication rate, because wound healing requires angiogenesis and fibroplasia as well as white blood cell activity, all of which are jeopardized. Hyperbaric oxygen therapy (HBO) raises oxygen levels in hypoxic tissue, stimulates angiogenesis and fibroplasia, and has antibacterial effects. METHODS: In this consecutive retrospective study, 15 patients with soft-tissue wounds without signs of healing after surgery in full-dose (64 Gy) irradiated head and neck regions were treated with HBO and adjuvant therapy. The patients in this study were also compared with patients examined in an earlier study, with corresponding wounds treated without HBO. RESULTS: The healing processes seemed to be initiated and accelerated by HBO. In the HBO group, 12 of 15 patients healed completely, 2 patients healed partially, and only 1 patient did not heal at all. There were no life-threatening complications. In the reference group, only 7 of 15 patients with corresponding wounds without signs of healing eventually healed without surgical intervention, and 2 patients had severe postoperative hemorrhage, which in one case was fatal. CONCLUSION: Evaluation of obtained results supports the hypothesis that HBO therapy has a clinically significant effect on initiation and acceleration of healing processes in irradiated soft tissues. PMID- 9213111 TI - Prognostic determinants in supraglottic carcinoma: univariate and Cox regression analysis. AB - BACKGROUND: A series of 281 consecutive patients affected by supraglottic cancer and treated with surgery alone or with surgery followed by radiotherapy between 1983 and 1989 was reviewed to identify significant prognostic determinants. METHODS: Fifty-one variables (related to host, tumor, and treatment) were tested by univariate and multivariate analysis performed on absolute and determinate survival. RESULTS: The final model of the multivariate analysis for absolute survival included the following covariates listed in order of higher relative risk of death: extracapsular spread, involvement of the medial wall of the pyriform sinus, thyroid cartilage invasion, metachronous tumor, anesthesiologic risk according to the American Society of Anesthesiologists classification (chi 2 = 71.28 with 6 d.f., p < .00001). The definitive model for determinate survival included: extracapsular spread, involvement of the medial wall of the pyriform sinus, extralaryngeal soft tissue invasion, and thyroid cartilage invasion (chi 2 = 82.74 with 5 d.f., p < .0001). CONCLUSIONS: Extracapsular spread was the most important factor affecting the prognosis of patients with supraglottic carcinoma. A second important finding was that T and N category did not emerge as a significant independent prognostic predictor at multivariate analysis. The negative impact on absolute survival of physical status and metachronous tumor could be the expression of the influence of concomitant diseases on survival. These observations concur to reinforce the concept that the current TNM classification is rather inadequate in predicting the prognosis of patients with supraglottic carcinoma when the aforementioned variables are considered. PMID- 9213113 TI - Jacob Da Silva Solis-Cohen: America's first head and neck surgeon. AB - Jacob Da Silva Solis-Cohen was a Renaissance physician and healer, who possessed great technical and intellectual skills. He was a self-made specialist in aerodigestive tract disease at a time when this was generally perceived as charlatanism. Unlike his contemporaries in laryngology, he was a trained surgeon, and he believed strongly that the laryngologist should be capable of doing his own surgery. Solis-Cohen was not a general surgeon who did work in the head and neck, but rather the first general surgeon who became a specialist in diseases of the upper air and food passages. His educational development is the model by which today's conventional paradigm in the training of Head and Neck Surgeons is designed. He fathered a medical and surgical specialty in America from its infancy in the 19th century until it was firmly grounded in the beginnings of this century. Just as Mosher was known as the "Chief" in the first half of this century, Jacob Da Silva Solis-Cohen had been designated the "Nestor" and "Chief" in laryngology many decades earlier. He died 2 months shy of 90 years on December 22, 1927. PMID- 9213112 TI - A re-evaluation of imaging criteria to assess aggressive masticator space tumors. AB - PURPOSE: To evaluate the correlation between the gross imaging evidence of an aggressive masticator space (MS) tumor and the presence of such a MS malignancy. MATERIALS AND METHODS: Thirty patients were identified retrospectively who had a malignancy that either arose in or metastasized to the MS, had pathologic verification of the diagnosis, and had magnetic resonance (MR) and/or computed tomographic (CT) images. Specifically evaluated was the presence or absence of gross imaging evidence of mandibular erosion and the integrity of the medial MS fascia as evaluated by a smooth margin between this fascia and the parapharyngeal space fat. RESULTS: Of the 30 tumors, 28 were high-grade malignancies and 2 were histiocytoses. Of these, 5 had mandibular erosion and violation of the MS fascia, 19 had bone erosion with an intact fascia, 4 had neither bone erosion nor fascial violation (3 of these patients were under the age of 20 years), and 2 had fascial violation with no bone erosion. CONCLUSIONS: In 76.7% of patients with a malignancy arising in the MS, on imaging the medial MS fascia was grossly intact. There were 4 patients with MS malignancy and neither violation of the medial MS fascia nor mandibular bone erosion. Thus, these imaging findings may not be good criteria to evaluate the presence of a high-grade MS malignancy, especially if the patient is under the age of 20 years, in which age group MS sarcomas are more likely to arise. PMID- 9213114 TI - Sphenoid sinus localization of multiple myeloma revealing evolution from benign gammopathy. AB - BACKGROUND: Plasma-cell neoplasms of the head and neck include extramedullary plasmacytoma and solitary plasmocytoma of bone or may represent a local manifestation of multiple myeloma. Involvement of sphenoid sinus has been rarely reported in multiple myeloma. METHODS: We present the case of a 77-year-old man with a 3-year-history of benign monoclonal IgG-lambda gammopathy who developed left sixth-nerve palsy and malaise. RESULTS: Computed tomography scan and magnetic resonance imaging scan disclosed a large soft-tissue mass of the sphenoid sinus with bone destruction. Sphenoid sinus biopsy revealed an IgG monoclonal plasma cell neoplasm. Diagnosis of multiple myeloma stage IA was then established. CONCLUSIONS: Diagnosis of plasma-cell neoplasm should be considered in sphenoid sinus tumors and depends upon histologic examination. This case enlightens the relationships between monoclonal benign gammopathy and plasma-cell neoplasms of the head and neck which constitute a continuum of B-cell lymphoproliferative disorders. PMID- 9213116 TI - Evidence for separate processing in the human brainstem of interaural intensity and temporal disparities for sound lateralization. AB - Sound lateralization can be induced by interaural intensity disparities (IIDs) or by interaural temporal disparities (ITDs). The purpose of this study was to indicate whether IIDs and ITDs are processed by the same central units that detect interaural disparity in timing of afferent activity. If sound lateralization to intensity and time cues was determined by the same afferent latency disparity detectors in the brainstem, lateralization would be the same, regardless of whether latency disparity was induced by IIDs or ITDs. Moreover, the disparity detectors, and thus their dipole equivalents, would be the same for equal lateralizations, whether induced by IIDs or ITDs. Auditory brainstem evoked potentials (ABEPs) were recorded in response to monaural and binaural clicks, with a variety of IIDs and ITDs. Peak II (proximal auditory nerve activity), peak III (input to the superior olivary complex), and binaural interaction components (BICs) BeI and BeII (binaurally activated upper pons) were identified and their latencies measured. The psychophysical lateralization of the clicks (in cm from vertex) was also measured in response to the same binaural stimuli. The correlations between interaural afferent latency disparities (difference in corresponding peak latencies originating in each ear) and psychophysical click lateralization were calculated. Similarly, the correlations with click lateralization of the BICs equivalent dipole latency as well as orientation change (relative to symmetrical clicks) were determined. A strong correlation with lateralization was found for peaks II and III latency disparities, with steeper slopes for IIDs than for ITDs. Moreover, binaural activity across the same lateralizations differed between IIDs and ITDs. These results, therefore, indicate that interaural time and intensity cues are processed by separate systems in the brainstem, both at the afferent convergence level and after interaural disparities are determined. PMID- 9213115 TI - Granulocyte colony stimulating factor-producing tongue carcinoma. AB - BACKGROUND: Leukocytosis without infection in patients with malignancies is known as the leukemoid reaction. The mechanisms involved in this phenomenon remain uncertain. METHODS: We describe the clinical, biochemical and immunohistochemical findings in a patient with recurrent tongue carcinoma accompanied by marked leukocytosis as high as 96200/ mm3. RESULTS: The serum granulocyte colony stimulating factor (G-CSF) concentration was increased to 204 (normal: < 30) pg/ml, which paralleled to the elevation of white blood cell (WBC) count and the tumor growth. The G-CSF content of the tumor tissue was also elevated (131 pg/mg protein) compared to that in control patients (6.63 +/- 2.63 pg/mg protein). Production of G-CSF from the tumor was evidenced by immunohistochemical staining with monoclonal antibody against human recombinant G-CSF. CONCLUSIONS: We suggest that the G-CSF production of the tumor participates in the mechanisms of the leukemoid reaction. PMID- 9213118 TI - Frequency-temporal resolution of hearing measured by rippled noise. AB - Frequency-temporal resolution of hearing was measured in normal hearers using rippled noise stimulation in conjunction with a phase-reversal test. The principle of the test was to interchange peak and trough positions (the phase reversal) and to find the highest ripple density at which such interchange is detectable depending on reversal rate. The measurements were made using narrow band noises with center frequencies of 0.5-4 kHz. The ripple-density resolution limits were constant at phase-reversal rates below 2-3/s and diminished at higher phase-reversal rates. A model is proposed to explain the data based on the envelope fluctuations inherent in noise; these fluctuations are supposed to limit detection of frequency-temporal sound patterns. PMID- 9213117 TI - Decline in the endocochlear potential corresponds to decreased Na,K-ATPase activity in the lateral wall of quiet-aged gerbils. AB - The ion transport-mediating enzyme, Na,K-ATPase, is abundantly present in the cochlear lateral wall. This enzyme is essential for the generation and maintenance of the endocochlear potential. Diminished enzyme activity has been observed previously in the lateral wall of quiet-aged gerbils. The present study was designed to investigate the impact of the age-related decline in Na,K-ATPase specific activity upon auditory function. Measures of the resting endocochlear potential value and the level of Na,K-ATPase specific activity were made in cochleae obtained from gerbils aged in quiet conditions. Analysis revealed a high degree of correspondence between the level of lateral wall Na,K-ATPase specific activity and the value of the endocochlear potential measured in the round window/turn 1 region of the cochlea. Nonlinear regression models showed a strong relationship between the age-related reductions in enzyme activity and the magnitude of the endocochlear potential. The data suggest that during metabolic presbyacusis a decrease in Na,K-ATPase specific activity can explain most, but not all, of the decline in the endocochlear potential. PMID- 9213119 TI - Effect of olivocochlear bundle section on evoked otoacoustic emissions recorded using maximum length sequences. AB - Presenting clicks according to maximum length sequences (MLS) enables transient evoked otoacoustic emissions (TEOAE) to be recorded at very high stimulation rates. As the click rate is increased from 40 clicks/s up to a maximum rate of 5000 clicks/s there is a reduction in TEOAE amplitude that reaches an approximate asymptote at 1500 clicks/s. One hypothesis put forward to explain this MLS 'rate effect' is that ipsilateral efferent activity is involved. To test this hypothesis TEOAEs were recorded from both ears of five patients who had undergone a unilateral vestibular nerve section--a surgical procedure which also entails sectioning the olivocochlear bundle. TEOAEs were recorded conventionally at 40 clicks/s and using MLS stimulation at 5000 clicks/s. Increasing the rate from 40 to 5000 clicks/s was found to reduce the amplitude of the TEOAEs by equivalent amounts in ears ipsilateral and contralateral to a vestibular nerve section as well as in the ears of normal-hearing adults. Since an ear ipsilateral to a vestibular nerve section should have no efferent innervation the hypothesis that efferent activity is the major mechanism involved in the MLS rate effect is rejected. Instead, the possibility that intracochlear processes are the underlying mechanism will now be investigated. PMID- 9213120 TI - Frequency summation observed in the human acoustic reflex. AB - It is known that the threshold of an acoustically induced middle-ear-muscle (MEM) reflex can be lowered by the simultaneous presentation of a second tone (facilitator), which is presented to the ipsilateral or contralateral ear at a level below the acoustic reflex threshold (ART) of the facilitator itself (Sesterhenn and Breuninger, 1976; Blood and Greenberg, 1981). In the present study, a primary elicitor and a facilitator were presented to the ear contralateral to that used for measurement of the acoustic reflex (AR), and the effects of changing frequencies and sound levels of the facilitator were investigated in human subjects with normal ears. The sound levels of facilitators, which caused a significant reduction of ART for the primary elicitors (facilitation thresholds), showed an asymmetrical pattern as a function of frequency of the facilitators. The facilitation thresholds tended to be lower when a facilitator with a frequency lower than the frequency of the elicitor (1 kHz) was used. In addition, effects of the elicitor on the masked thresholds of the facilitator were examined to observe the possible interaction between elicitor and facilitator from the viewpoint of 'spread of excitation'. The underlying mechanism of summation effects of two tones are discussed based on the possible input mechanism involved in the acoustically induced MEM reflex are. PMID- 9213121 TI - Synaptic specializations associated with the outer hair cells of the Japanese macaque. AB - Across species the innervation of outer hair cells (OHCs) shows a remarkable similarity. There are, however, notable differences in fine structure. The present work describes the normal synaptic morphology of OHCs in the Japanese macaque (Macaca fuscata), as determined by examination of serial sections with transmission electron microscopy. The nerve endings at the base of OHCs were divided primarily into two groups: vesiculated (efferent) and non-vesiculated (afferent). In addition, we found supranuclear efferent nerve endings and reciprocal synapses in all three cochlear turns. We also found presynaptic bodies in OHCs at the afferent synapse, the branching of afferent nerve fibers and axodendritic synapses between afferent and efferent fibers in the outer spiral bundle and just beneath OHCs. In terms of synaptic structure, the data indicate that that the Japanese macaque is more similar to that of the human than other species examined to date. PMID- 9213122 TI - Focal microcirculation disorder induced by photochemical reaction in the guinea pig cochlea. AB - A small region of microcirculation disorder in the cochlea of the guinea pig could be induced by a photochemical reaction. Photoillumination to the cochlea was done after systemic infusion of Rose Bengal (RB). The lateral wall of the second or third turn of the cochlea was illuminated for 10 min with a 1 mm diameter focused green light supplied by a xenon lamp. Degeneration of the stria vascularis (SV) was observed by a scanning electron microscope at 60-300 min after illumination. The range of length of degenerated area in the SV was from 111 to 1800 microns, with a mean of 760 microns. The organ of Corti along the illuminated lesion of the SV was well preserved in all animals at 60-300 min. In contrast, degeneration of sensory hair cells and scar formation in the SV were observed in the focal lesions of the three animals killed 1 week after illumination. The increase of diameter in the vessel of the SV from the radiating arteriole, the vessel of basilar membrane (VSBM) and limbus vessel (LVS) were observed in the illuminated area with diaminobenzidine (DAB) staining. These findings suggest that segmental microcirculation damage occurred in the SV and modiolus. In physiological studies, compound action potentials (CAP) were evaluated. Endocochlear potentials (EP) were also measured at the second turn under three different situations (groups A, B and C). A photochemically induced lesion was created at the site of EP measurement (group A), a site in the second turn 1 mm from the EP measurement site (group B) and a site in the third turn adjacent to the EP measurement site (group C). Threshold shift of CAP (up to 5.6 +/- 1.8 dB SPL) and reduction of EP (down to 11.4 +/- 10.7 mV) in the photochemically injured location were detected during about 15 min. EP did not recover to the predamaged level (79.9 +/- 3.7 mV) during 20 min. The morphological and physiological changes were not observed in the control group with illumination only. There were no significant decreases in EP values at the sites 1 mm from the lesion (group B) and at the inferior turn adjacent to the lesion (group C) compared to the marked decrease at the site of the photochemically induced lesion (group A). These findings suggest that CAP and EP are significantly affected by the interruption of segmental blood supply in the cochlea and remarkable decrease of EP occurs in the focal region of the guinea pig cochlea. We conclude that a localized blood circulation disorder induced by the photochemical reaction can make a focal lesion in guinea pig cochlea morphologically and physiologically. PMID- 9213123 TI - Nitric oxide synthase and contractile protein in the rat cochlear lateral wall: possible role of nitric oxide in regulation of strial blood flow. AB - The present study demonstrated by histochemical and immunohistochemical methods that NADPH diaphorase reactivity, endothelial nitric oxide synthase (eNOS)-like immunoreactivity, and tropomyosin-like immunoreactivity, were located within the rat cochlear lateral wall. Both NADPH diaphorase reactivity and eNOS-like immunoreactivity were found mainly in the endothelium of the strial capillaries (ESC) and that of the vessels of the spiral ligament (ESL). These reaction products appeared to be somewhat more common in the ESC than in the ESL. On the other hand, tropomyosin-like immunoreactivity was localized in tissues outside the endothelium and its intensity was greater in the ESL than in the ESC. These findings suggest that nitric oxide (NO) produced by eNOS may play a role in regulating the blood flow of the cochlear lateral wall. In addition, NADPH diaphorase reactivity, eNOS-like immunoreactivity, and tropomyosin-like immunoreactivity showed different patterns of distribution between ESC and ESL. This suggests that in these two sites blood circulation is controlled by NO through two different mechanisms that are suitable for regulating strial blood flow. PMID- 9213124 TI - Morphologic evidence for innervation of Deiters' and Hensen's cells in the guinea pig. AB - The presence of nerve fibers and terminals among Deiters' and Hensen's cells of the organ of Corti of the adult guinea pig is demonstrated using immunostaining for synaptophysin and neurofilaments, acetylcholinesterase histochemistry, and transmission electron microscopy. These nerve terminals appeared to form chemical synapses with Deiters' and Hensen's cells. Nerve fibers and synapses were more common in the apical as compared to the basal cochlea. The terminals were often present on basal appendages of Hensen's cells, which were rich in mitochondria and often contained a Golgi apparatus and dense core vesicles. Electron microscopy and immunostaining for neurofilaments showed that most Hensen's cells in the apical cochlea received innervation. Few of the nerve fibers and terminals were positive for acetylcholinesterase, which suggests that they were not collaterals of cholinergic olivocochlear fibers. The density of these fibers, as shown by immunohistochemistry for neurofilaments, was far greater than previous reports of GABA-ergic fibers, which suggests that they were not GABA-ergic olivocochlear fibers. The role of such fibers and synapses with supporting cells of the outer hair cell area is unknown. Determination of the origins and functions of these fibers will provide new insights into cochlear structure and function. PMID- 9213125 TI - Characteristics of DPOAE audiogram in tinnitus patients. AB - To investigate cochlear activity in tinnitus, the DPOAE (distortion product otoacoustic emission) audiograms (DP-gram) of tinnitus patients were measured. Nine tinnitus patients (15 ears) with normal hearing and 55 tinnitus patients (75 ears) with hearing impairment were included in this study. Significant decreases in DPOAE amplitude over a limited frequency range were observed in 93.3% of the normal hearing tinnitus group and in 96% of the hearing-impaired tinnitus group. The averaged DP-gram of the normal hearing tinnitus group was significantly different from that of the normal subject (repeated-measures ANOVA, P < 0.01). These results imply that tinnitus may be evaluated objectively by DPOAE. PMID- 9213126 TI - Efferent innervation of the inner hair cell region: origins and terminations of two lateral olivocochlear systems. AB - The projections of lateral olivocochlear neurons (LOC), which terminate beneath inner hair cells (IHCs), were investigated by injecting biotinylated dextran amine into the lateral superior olivary nucleus (LSO) and the surrounding region in the rat. This region has been definitively shown to contain two types of olivocochlear neurons: small cells within the LSO (intrinsic neurons) and large cells (shell neurons) surrounding it (Vetter, D.E., Mugnaini, E., 1992. Distribution and dendritic features of three groups of rat olivocochlear neurons. Anat. Embryol. 185, 1-16). Labeled efferent axons were studied by light microscopy in whole mounts and radial sections of the organ of Corti (OC). It was found that injections confined to the LSO, which presumably affected mainly intrinsic neurons, labeled a cluster of axons in the osseous spiral lamina that entered the inner spiral bundle (ISB) and terminated in one or more dense patches that, in total basal-apical extent, spanned no more than 10-20% (1-2 mm) of the total length of the OC (10 mm). In contrast, injections affecting shell neurons produced labeled axons that entered the OC over a span of more than 50% of its length and which, as a group, coursed in the ISB for at least 80%, and sometimes more than 95% of total cochlear length. Study of individual axons in the OC revealed that intrinsic axons did not bifurcate upon entering the OC and traveled less than 1 mm before terminating in a discrete, dense arbor. In contrast, shell axons typically bifurcated into basal and apical branches that, in toto, traveled between 1 and 2 mm beneath the IHCs, forming numerous en passant swellings and a few terminal branches en route. The fact that localized injections of intrinsic neurons produced focal peaks of labeling in the cochlea, whereas similar injections of shell neurons produced a diffuse, non-focal projection that could extend for nearly the entire length of the cochlea, suggests that significant differences exist between these two populations in their capacity to influence localized, frequency-specific regions of the OC, and thus in their probable functional roles. The present findings in the rat not only confirm a previous study in the guinea pig which found a similar dual efferent innervation beneath the IHCs (Brown, M.C., 1987. Morphology of labeled efferent fibers in the guinea pig cochlea. J. Comp. Neurol. 260, 605-618), but extend those observations by linking two axonal types beneath the IHCs to their respective cell bodies of origin in the lateral zone of the superior olivary complex. PMID- 9213127 TI - Electrical stimulation of the auditory nerve. I. Correlation of physiological responses with cochlear status. AB - The purpose of the present study was to evaluate evoked potential and single fibre responses to biphasic current pulses in animals with varying degrees of cochlear pathology, and to correlate any differences in the physiological response with status of the auditory nerve. Six cats, whose cochleae ranged from normal to a severe neural loss (< 5% spiral ganglion survival), were used. Morphology of the electrically evoked auditory brainstem response (EABR) was similar across all animals, although electrophonic responses were only observed from the normal animal. In animals with extensive neural pathology, EABR thresholds were elevated and response amplitudes throughout the dynamic range were moderately reduced. Analysis of single VIIIth nerve fibre responses were based on 207 neurons. Spontaneous discharge rates among fibres depended on hearing status, with the majority of fibres recorded from deafened animals exhibiting little or no spontaneous activity. Electrical stimulation produced a monotonic increase in discharge rate, and a systematic reduction in response latency and temporal jitter as a function of stimulus intensity for all fibres examined. Short-duration current pulses elicited a highly synchronous response (latency < 0.7 ms), with a less well synchronized response sometimes present (0.7 1.1 ms). There were, however, a number of significant differences between responses from normal and deafened cochleae. Electrophonic activity was only present in recordings from the normal animal, while mean threshold, dynamic range and latency of the direct electrical response varied with cochlear pathology. Differences in the ability of fibres to follow high stimulation rates were also observed; while neurons from the normal cochlea were capable of 100% entrainment at high rates (600-800 pulses per second (pps)), fibres recorded from deafened animals were often not capable of such entrainment at rates above 400 pps. Finally, a number of fibres in deafened animals showed evidence of 'bursting', in which responses rapidly alternated between high entrainment and periods of complete inactivity. This bursting pattern was presumably associated with degenerating auditory nerve fibres, since it was not recorded from the normal animal. The present study has shown that the pathological response of the cochlea following a sensorineural hearing loss can lead to a number of significant changes in the patterns of neural activity evoked via electrical stimulation. Knowledge of the extent of these changes have important implications for the clinical application of cochlear implants. PMID- 9213128 TI - Multi-unit mapping of acoustic stimuli in gerbil inferior colliculus. AB - Multi-unit peristimulus time (MU-PST) histograms were recorded in the gerbil inferior colliculus (IC) in response to tone burst stimuli. Histograms were collected every 100 microns as the recording electrode was advanced along the tonotopic axis of the central nucleus of the IC. Space/time maps of neural activity were constructed from these data. In most of our sample the pattern of response changed systematically as the stimulating frequency was increased in octave steps. At low frequencies (< 500 Hz) the pattern of response was broadly distributed spatially and phase-locked to the stimulus frequency. At higher frequencies (> 1 kHz) the pattern of response was more localized and showed no evidence of phase locking. The location of the maximum response to tones from 1 to 32 kHz moved ventrally along the tonotopic axis at an approximate rate of 230 microns/stimulus octave. The patterns of response were localized near stimulus threshold and spread over a larger region as level increased. This method of collecting and displaying multi-unit response maps provides an overview of ensemble activity that allows concurrent observation of spatial and temporal variations in activity patterns. The quantitative analysis of components of MU PST Maps are consistent with trends illustrated with single-unit tuning and level functions. This perspective of IC activity suggests potential processing mechanisms that are congruent with single-unit reconstructions. PMID- 9213130 TI - The postnatal development of F-actin in tension fibroblasts of the spiral ligament of the gerbil cochlea. AB - The tension fibroblasts of the spiral ligament of the mammalian cochlea are thought to create radial tension on the basilar membrane. Their postnatal development was investigated in the gerbil (Meriones unguiculatus) with confocal fluorescence microscopy using phallotoxin as a specific marker for F-actin. In the adult cochlea, tension fibroblasts were restricted to the basal cochlear turn and were arranged in 2-4 rows in the marginal region of the spiral ligament. They contained intensely stained parallel bundles of F-actin. In upper cochlear turns, the marginal region of the spiral ligament was occupied by sparsely distributed, unobtrusively labeled fibrocytes, the bone lining cells. The spiral ligament of young postnatal stages (newborn--6 days after birth (DAB)) lacked F-actin labeling patterns that are characteristic for tension fibroblasts in the adult. Rather, the whole inner surface of the otic capsule throughout all cochlear turns was outlined by cell layers with distinct but diffuse cytoplasmic F-actin label. These cells may represent perichondrial fibrocytes. Around 9 DAB, the perichondrium revealed changes in morphology and F-actin patterns that indicate a further differentiation into tension fibroblasts (basal turn) or bone lining cells (more apical turns). At 12 DAB, around onset of hearing, adult-like bone lining cells were found in the marginal regions of the spiral ligament of upper cochlear turns. In the basal turn, tension fibroblasts were present, but their F actin cytoskeleton was not fully developed. During the following days, F-actin label increased in tension fibroblasts and reached adult-like configuration at 17 DAB, coinciding with mature hearing characteristics. The role of tension fibroblasts in development of hearing characteristics is discussed. PMID- 9213129 TI - Action potentials and underlying voltage-dependent currents studied in cultured spiral ganglion neurons of the postnatal gerbil. AB - The excitability of cultured spiral ganglion (SG) neurons from early postnatal gerbil (P0-P1) was examined with the whole-cell patch-clamp technique. The role of voltage-gated currents in shaping the kinetics of action potentials (APs) was analyzed. Cultured SG neurons displayed spontaneous APs with a low rate (< 0.1 Hz). The kinetics of APs were studied by injecting neurons with current pulses of various frequencies and duration. A single depolarizing pulse of long duration elicited only one AP in most SG neurons. When excited by a train of short current pulses given at rates greater than 50 Hz, the firing pattern displayed an adaptive mechanism with the result that successive APs fired with lower amplitude, broader duration and delayed peak time. Pulse trains of higher frequencies had higher failure rates in initiating APs. Current pulses given at 20 Hz or lower elicited APs that had very similar amplitudes. However, the width of the APs gradually broadened. Duration of APs was also found to be affected by the membrane potential of neurons. Between -75 mV and -55 mV, AP duration was broadened at a rate of about 33% per 10 mV depolarization. Voltage-gated currents that underlie the generation of APs were examined under voltage-clamp conditions. Tetrodotoxin-sensitive sodium currents and dihydropyridine-sensitive L-type calcium currents were found. More importantly, inactivation properties of the potassium current provided a direct explanation for the cumulative broadening of APs. This work demonstrated that SG neurons were able to fire APs long before hearing commences in gerbil. Possible roles of spontaneous APs in the development of the cochlea and the role of voltage-gated currents in the function of SG neurons under normal and pathological conditions are discussed. PMID- 9213131 TI - Low-frequency modulation of inner hair cell and organ of Corti responses in the guinea pig cochlea. AB - Low-frequency tones are used to study changes in responsiveness as a function of phase in inner hair cell (IHC) and organ of Corti (OC) responses recorded from second turn of the guinea pig cochlea. In these experiments a 40 Hz stimulus is combined with a variable frequency probe to determine the degree to which tones at and below best frequency (BF) are modulated. Changes in responsiveness produced by the low-frequency input are quantified and related to position of the basilar membrane which is estimated using the phase of the cochlear microphonic measured in the OC fluid space. Results obtained when 40 Hz is presented at its lowest effective level demonstrate that ac responses to low-level BF probes are reduced for basilar membrane displacements to scala tympani while probe tones well below BF are modulated in the opposite direction. The transition between these two response patterns occurs when the overall DC produced in the OC by the two-tone input changes from positive to negative. Because of this association, the frequency dependence exhibited in the bias results may be linked to mechanisms responsible for generating the two polarities of the summating potential and the DC receptor potentials that it reflects. An attempt is also made to relate bias-induced changes in hair cell receptor potentials to modulations in single-unit rate responses. In other words, to address variations in the temporal relationships between excitation and suppression measured in the auditory nerve. PMID- 9213132 TI - Using new genetics tools to advance the behavioral neurosciences beyond nature versus nurture. PMID- 9213133 TI - The use of genetic "knockout" mice in behavioral endocrinology research. AB - The production of mice with specific deletion of targeted genes (knockouts) has provided a useful tool in understanding the mechanisms underlying behavior. There are many opportunities with this new tool for behavioral neuroendocrinology, specifically, and behavioral biology, generally. Although this genetic technique offers new opportunities to study the mechanisms of behavior, as with all behavioral techniques there are some potential limitations. For example, the products of many genes are essential to normal function, and inactivating the gene may prove lethal or induce gross morphological or physiological abnormalities that can complicate interpretation of discrete behavioral effects. Unexpected compensatory or redundancy mechanisms might be activated when a gene is missing and cloud interpretation of the normal contribution of the gene to behavior. Behavioral tests study the effects of the missing gene (and gene product), not the effects of the gene directly. This conceptual shortcoming can be overcome in the same way that it is overcome in other types of ablation studies, by collecting converging evidence using a variety of pharmacological, lesion, and genetic manipulations. Finally, because mammalian genome mapping is currently focused on mice (Mus musculus), standardized behavioral testing of mice should be adopted. Against those disadvantages are several important advantages to using knockout mice in behavioral research: (1) disabling a gene is often a very precise and "clean" ablation, (2) the effects of the gene product can be abolished without the side-effects of drugs, and (3) genetic manipulations may be the only way to determine the precise role of many endogenous factors on behavior. The use of new inducible knockouts, in which the timing and placement of the targeted gene disruption can be controlled, will be an extremely important tool in behavioral endocrinology research. PMID- 9213134 TI - A proposed test battery and constellations of specific behavioral paradigms to investigate the behavioral phenotypes of transgenic and knockout mice. AB - Behavioral phenotyping of transgenic and knockout mice requires rigorous, formal analyses. Well-characterized paradigms can be chosen from the established behavioral neuroscience literature. This review describes (1) a series of neurological and neuropsychological tests which are effectively used as a first screen for behavioral abnormalities in mutant mice, and (2) a series of specific behavioral paradigms, clustered by category. Included are multiple paradigms for each category, including learning and memory, feeding, analgesia, aggression, anxiety, depression, schizophrenia, and drug abuse models. Examples are given from the experiences of the authors, in applying these experimental designs to transgenic and knockout mice. Extensive references for each behavioral paradigm are provided, to allow new investigators to access the relevant literature on behavioral methodology. PMID- 9213135 TI - What nature's knockout teaches us about GnRH activity: hypogonadal mice and neuronal grafts. AB - The hypogonadal mouse is one of "nature's knockouts," bearing a specific deletion in the gene for gonadotropin-releasing hormone (GnRH), with the result that no GnRH peptide is detectable in the brain. The lack of reproductive development after birth provides an animal model that has proved fruitful in clarifying the role of GnRH in reproductive behavior and physiology. Behavioral studies with hypogonadal mice convincingly demonstrate that although GnRH may facilitate the appearance of sexual behavior, this peptide is not essential for either male or female sexual behavior in the mouse. Administration of GnRH to hypogonadal mice with regimens mimicking GnRH pulsatility initiates reproductive development. Surprisingly, continuous exposure to GnRH stimulates remarkable ovarian and uterine growth and increased FSH release, although pituitary content of LH and FSH remains unchanged. In contrast, when brain grafts of normal fetal preoptic area (POA), containing GnRH cells, are implanted in the third ventricle of adult hypogonadal mice, both pituitary and plasma gonadotropin levels increase. Grafted GnRH neurons innervate the median eminence of the host and support pulsatile LH secretion in the majority of animals with graft-associated gonadal development. Studies of hypogonadal mice with POA grafts demonstrate that distinct components of reproductive function are dissociable: hosts may demonstrate reflex but not spontaneous ovulation; others may show positive but not negative feedback. Activation of grafted GnRH cells in response to sensory input to the host, as revealed in Fos expression studies, is an example of the integration of the graft with the host brain that underlies such capabilities. A goal of these studies is to elucidate the specific connectivity underlying discrete aspects of reproductive function. PMID- 9213136 TI - Gene targeting approaches to neuroendocrinology: oxytocin, maternal behavior, and affiliation. AB - Transgenic technology affords exciting new opportunities in the field of behavioral neuroendocrinology. We have extended our research into the behavioral function of oxytocin in maternal and social behavior using two transgenic approaches: (i) targeted deletion of the oxytocin gene in mice and (ii) augmented oxytocin receptor expression in the brain. Mice genetically deficient in oxytocin can mate, give birth, and display normal maternal behavior; however, milk ejection and certain aspects of social behavior are affected. Comparative studies of oxytocin receptors have led to the observation that species differences in social organization are associated with differences in receptor distribution. Specifically, monogamous prairie voles and nonmonogamous, asocial montane voles exhibit different patterns of OT receptor expression in the brain. Transgenic mice have been created with a reporter gene driven by the prairie vole oxytocin receptor gene promoter. Analysis of the expression pattern suggests that it should be possible to manipulate receptor expression in the vole brain in order to examine the effects of receptor distribution on behavior. PMID- 9213137 TI - Estrogen receptor function as revealed by knockout studies: neuroendocrine and behavioral aspects. AB - Estrogens are an important class of steroid hormones, involved in the development of brain, skeletal, and soft tissues. These hormones influence adult behaviors, endocrine state, and a host of other physiological functions. Given the recent cloning of a second estrogen receptor (ER) cDNA (the ER beta), work on alternate spliced forms of ER alpha, and the potential for membrane estrogen receptors, an animal with a null background for ER alpha function is invaluable for distinguishing biological responses of estrogens working via the ER alpha protein and those working via another ER protein. Data generated to date, and reviewed here, indicate that there are profound ramifications of the ER alpha disruption on behavior and neuroendocrine function. First, data on plasma levels of estradiol (E2), testosterone (T), and luteinizing hormone (LH) in wild-type (WT) versus ER alpha- mice confirm that ER alpha is essential in females for normal regulation of the hypothalamic-pituitary gonadal axis. Second, ovariectomized female ER alpha- mice do not display sexual receptivity when treated with a hormonal regime of estrogen and progesterone that induces receptivity in WT littermates. Finally, male sexual behaviors are disrupted in ER alpha- animals. Given decades of data on these topics our findings may seem self-evident. However, these data represent the most direct test currently possible of the specific role of the ER alpha protein on behavior and neuroendocrinology. The ER alpha- mouse can be used to ascertain the specific functions of ER alpha, to suggest functions for the other estrogen receptors, and to study indirect effects of ER alpha on behavior via actions on other receptors, neurotransmitters, and neuropeptides. PMID- 9213138 TI - Progesterone receptor function from a behavioral perspective. AB - Hormonal induction of sexual receptivity in ovariectomized female mice can be effectively reinstated by sequential administration of estradiol and progesterone. In this regard, mice appear to be similar to other rodents. While it is generally accepted that hypothalamic progesterone receptors function as estradiol-induced transcription factors in the induction of sexual receptivity in rats, hamsters, and guinea pigs, relatively little is known about their role in the mouse, a species which exhibits genotypic and strain differences in the responsiveness to steroid hormones. Using a transgenic mouse carrying a null mutation for the progesterone receptor by gene targeting, we examined the role of the progesterone receptor as a coordinator of key regulatory events in the induction of sexual receptivity. A concordance between hypothalamic progesterone receptor levels and behavioral responsiveness was established by comparing the homozygous mutant, heterozygous mutant, and wild-type littermates. The behavioral and biochemical findings reveal the importance of estradiol-induced progesterone receptors for the expression of sexual behavior in female mice. The behavioral response of the two parental mouse strains from which the recombinant genotype was generated was also examined. As an extension of our earlier studies on the ligand-independent activation of progesterone receptors by neurotransmitters, the behavioral effect of dopamine in the facilitation of sexual receptivity in mice was also examined. The studies provide further evidence that steroid hormone receptors function as general transcription factors to achieve the integration of neural information in the central nervous system, and they assign a more important role for progesterone receptors than hitherto envisioned. PMID- 9213139 TI - Pheromone-induced spawning of Pacific herring. I. Behavioral characterization. AB - A spawning pheromone in the milt (semen) and testes of the Pacific herring, Clupea harengus pallasi, triggers spawning in sexually mature fish of both sexes and is thought to facilitate school spawning of this species. We found the response to the pheromone to be a stereotyped behavioral sequence consisting of a graded extension of the gonadal papilla, release of gametes, and spawn deposition behavior. The response is triggered by an olfactory stimulus, as demonstrated by the elimination of the response by occlusion of the nares. Stimulus concentrations of an approximate 1:500 dilution of fresh milt or the equivalent of 0.02 g of fully mature testes per milliliter were required to elicit a response in 50% of ripe herring that are responsive to the pheromone. Female fish appeared to be less sensitive to the pheromone in milt than males early in the spawning season, but not thereafter. The average duration of responses of male fish was longer after exposure to concentrated milt than to testes extracts, but no consistent difference in response times of the two sexes was detected. Factors other than the spawning pheromone, maturity of the fish, and stress also were found to influence the spawning response. For example, exposure to shallow (3 cm) water in a small tank induced "spontaneous" papilla extension and spawning approximately 20 min after refilling the tank; occluding the nares prevented this response. Also, the presence of floating kelp (Macrocystis) resulted in prolonged spawning in a large tank after pheromonal stimulation. PMID- 9213140 TI - Pheromone-induced spawning of Pacific herring. II. Plasma steroids distinctive to fish responsive to spawning pheromone. AB - A spawning pheromone in the milt (semen) and testes of the Pacific herring, Clupea harengus pallasi, is thought to facilitate school spawning of this species. We found that responsiveness to the spawning pheromone was variable among ripe fish (milt-producing or ovulated). Measurement of five principle reproductive steroids in the free form and five steroids in conjugated forms in the plasma of male fish early in the spawning season (newly ripe fish) showed that elevated plasma levels of 3 alpha, 17 alpha-dihydroxy-3 beta-pregnan-20-one and 17 alpha-hydroxyprogesterone coincided with responsiveness to the spawning pheromone in these fish; levels of other steroids did not differ. In contrast, responsiveness to the pheromone by female fish later in the spawning season (ripe and-holding fish) coincided with lower levels of glucuronated 17 alpha,20 beta dihydroxyprogesterone and a lower gonadosomatic index. We suggest that these differences indicate a more advanced mature reproductive state in the responsive individuals among both the newly ripe male and the ripe-and-holding female fish. We found no differences in the level of cortisol in the blood of the herring that could be correlated with differences in pheromonal responsiveness. We conclude that differences in responsiveness to the spawning pheromone coincide to some extent with levels of reproductive maturation but probably not with recent stress. PMID- 9213141 TI - Combined olfactory contact with the parent colony and direct contact with nonbreeding animals does not maintain suppression of ovulation in female naked mole-rats (Heterocephalus glaber). AB - The study investigated the role of odor cues from two naked mole-rat colonies, in conjunction with behavioral cues from nonbreeding colony members, in maintaining suppression of ovulation in subordinate female naked mole-rats isolated from the two parent colonies. Four high ranking nonbreeding female naked mole-rats were removed from their respective parent colonies and singly housed in separate burrow systems. For a 64-day period, the removed females were maintained in daily odor contact with their parent colony by daily rotating soiled bedding material between the parent colony and the burrow systems of removed females. In addition, subsets of nonbreeding animals from the respective parent colony were regularly moved into the burrow systems of removed females for 2-day periods during this 64 day period. Removed females were therefore in continual social contact with subsets of parent colony animals except for the breeding pair. All four removed females exhibited raised levels of urinary progesterone (< 2 ng/mg Cr) indicative of the onset of ovarian function within 3 days of being separated from the parent colony. Removed females exhibited a normal ovulatory cycle with levels of progesterone remaining elevated for 25-35 days (mean concentration of progesterone +/- SEM; 16.2 +/- 2 ng/mg Cr). Initiation of aggression and sexual behavior by removed females increased significantly when they were isolated from the parent colony. The results demonstrated that odor cues from the complete colony in conjunction with behavioral/tactile/vocal cues from the nonbreeding colony members were not the major cues maintaining reproductive suppression in nonbreeding female naked mole-rats. Instead, our results suggest that female reproductive suppression in naked mole-rats is caused by a dominance-related behavioral mechanism requiring direct contact with the breeding female. PMID- 9213142 TI - Male gray short-tailed opossums (Monodelphis domestica) receive penile intromissions when treated with estrogen and progesterone in adulthood. AB - Following treatment with estradiol and progesterone, gonadectomized male as well as female gray opossums received penile intromissions from intact stimulus males. Intromission was possible in male gray opossums because, like marsupials of both sexes, they possess a single cloaca-like anogenital opening. All subjects that allowed intromission showed anogenital dragging just prior to intromission. While intromission latency was similar in tests involving male and female subjects, total intromission duration was longer in tests involving male subjects than in tests involving female subjects, and sex locks were seen only in tests involving female subjects. These findings are discussed with respect to the potential usefulness of gray opossums for studying the effects of peripheral vs central factors on the display of sex differences in behavior. PMID- 9213143 TI - Behavioral dominance and corpus luteum function in red deer Cervus elaphus. AB - Times of the ovulatory LH surge and characteristics of the rise in circulating progesterone concentrations after ovulation in red deer hinds were investigated in relation to each animal's dominance status. Observations were made during the 1992 (experiment 1) and 1993 (experiment 2) breeding seasons, while the same 12 hinds were held in a pen in the absence of a stag. Ovulation was synchronized by administration of progesterone followed by luteolytic prostaglandin F2 alpha analogue. Social status was determined for each hind by noting dyadic agonistic interactions during the period of progesterone treatment. Hinds were weighed before and after the experiments. Time of onset of estrus (lordosis) was recorded while handling at 3-hr intervals (for 81 hr in experiment 1; 96 hr in experiment 2) after progesterone withdrawal, and blood samples were collected at these times to characterize the preovulatory LH surge. Subsequently, daily blood samples were collected for up to 11 days for measurement of progesterone concentrations. There was a tendency for weight change to be related to dominance status in experiment 1 and this was significant in experiment 2 (p < 0.01). The rate of increase in circulating progesterone concentration after ovulation was related to status (data of experiments 1 and 2 combined; p < 0.02), but was not correlated with the time of estrus or with the time or height of the LH surge. A third experiment (carried out in 1993), when the same hinds were kept with a stag after induced ovulation, showed that time of estrus (mating) was not related to dominance status. The data suggest that corpus luteum function is affected by social status. The results are discussed in the context of mechanisms by which dominance status influences the sex of a hind's calves. PMID- 9213145 TI - Epidemiology and injury prevention. PMID- 9213144 TI - Watching the canary: the prevention of suicide. PMID- 9213146 TI - Injury and entropy. PMID- 9213147 TI - Motor vehicle occupant protection: have we become too complacent? PMID- 9213149 TI - 'Accident proneness': statistical and practical significance. PMID- 9213148 TI - Is saying NO to 'accident proneness' throwing the baby out with the bathwater? PMID- 9213150 TI - President Clinton's radio address on child passenger safety. PMID- 9213151 TI - Statistical commentary. PMID- 9213153 TI - Measuring the implementation of injury prevention programs in state health agencies. AB - OBJECTIVE: Injury prevention programs have been implemented with varying degrees of success in the United States. The objective of this study was to identify the variables that influence the successful implementation of injury prevention programs. METHODS: The key indicators of implementation success and its correlates were identified through consultation with a panel of experts. This consultation informed the content of a mail questionnaire sent to all United States state health departments, followed by telephone interviews. Data were analyzed using factor analysis and regression to identify significant relationships between variables. RESULTS: Data were obtained from 64 programs, representing 44 states; these included 24 programs in injury control units, 12 in maternal and child health units, 10 in health promotion/education units; and eight in emergency medical services units. Analysis identified four factors that are associated with an index of successful injury prevention program implementation; (1) participation and advocacy by constituent groups; (2) organizational capacity; (3) administrative control; and (4) attributes of relevant policies. CONCLUSIONS: Findings indicated that constituent participation (the extent and efficacy of constituency support and advocacy) and organizational capacity (a function of program staff and their skill levels) had the greatest influence on successful program implementation. Support from advocacy groups and knowledgeable staff members, whose time is dedicated to the program, are critical for conducting the activities necessary for successful implementation of these programs. PMID- 9213152 TI - An international study of the exposure of children to traffic. AB - OBJECTIVES: To examine the extent of international differences in children's exposure to traffic as pedestrians or bicyclists. DESIGN: Children's travel patterns were surveyed using a parent-child administered questionnaire. Children were sampled via primary schools, using a probability cluster sampling design. SETTING: Six cities in five countries: Melbourne and Perth (Australia), Montreal (Canada), Auckland (New Zealand), Umea (Sweden), and Baltimore (USA). SUBJECTS: Children aged 6 and 9 years. MAIN OUTCOME MEASURES: Modes of travel on the school home journey, total daily time spent walking, and the average daily number of roads crossed. MAIN FINDINGS: Responses were obtained from the parents of 13423 children. There are distinct patterns of children's travel in the six cities studied. Children's travel in the three Australasian cities, Melbourne, Perth and Auckland, is characterised by high car use, low levels of bicycling, and a steep decline in walking with increasing car ownership. In these cities, over a third of the children sampled spent less than five minutes walking per day. In Montreal, walking and public transport were the most common modes of travel. In Umea, walking and bicycling predominated, with very low use of motorised transport. In comparison with children in the Australasian and North American cities, children in Umea spend more time walking, with 87% of children walking for more than five minutes per day. CONCLUSIONS: There are large international differences in the extent to which children walk and cycle. These findings would suggest that differences in 'exposure to risk' may be an important contributor to international differences in pedestrian injury rates. There are also substantial differences in pedestrian exposure to risk by levels of car ownership-differences that may explain socioeconomic differentials in pedestrian injury rates. PMID- 9213154 TI - A descriptive analysis of children's playground injuries in the United States 1990-4. AB - OBJECTIVES: To review playground injury statistics over a five year period in order to develop an awareness of how and where children in the United States are being injured. METHODS: All data are based on the United States Consumer Product Safety Commission's National Electronic Injury Surveillance System (NEISS) for playground related injuries during 1990-4. The surveillance data includes injuries recorded in more than 90 hospital emergency departments located throughout the United States. RESULTS: Each year there are roughly 211,000 preschool or elementary school-children in the United States who receive emergency department care for injuries associated with playground equipment. On average, 17 of these cases result in death. 70% of all injuries occur on public playgrounds, with nearly one third classified as severe. Swings, climbers, and slides are the pieces of playground equipment associated with 88% of all NEISS reported injuries. Falls to the surface are responsible for 70%. CONCLUSIONS: NEISS playground injury statistics contribute to our understanding of playground injuries. By identifying where and how children are injured, suggestions can be made in an attempt to make playgrounds safer. PMID- 9213155 TI - Reduction in paediatric burn admissions over 25 years, 1970-94. PMID- 9213156 TI - Epidemiology of bicycle injuries and risk factors for serious injury. AB - OBJECTIVE: To determine the risk factors for serious injury to bicyclists, aside from helmet use. DESIGN: Prospective case-control study. SETTING: Seven Seattle area hospital emergency departments and two county medical examiner's offices. PATIENTS: Individuals treated in the emergency department or dying from bicycle related injuries. MEASUREMENTS: Information collected from injured bicyclists or their parents by questionnaire on circumstances of the crash; abstract of medical records for injury data. Serious injury defined as an injury severity score > 8. ANALYSIS: Odd ratios computed using the maximum likelihood method, and adjusted using unconditional logistic regression. RESULTS: There were 3854 injured cyclists in the three year period; 3390 (88%) completed questionnaires were returned 51% wore helmets at the time of crash. Only 22.3% of patients had head injuries and 34% had facial injuries. Risk of serious injury was increased by collision with a motor vehicle (odds ratio (OR) = 4.6), self reported speed > 15 mph (OR = 1.2), young age (< 6 years), and age > 39 years (OR = 2.1 and 2.2 respectively, compared with adults 20-39 years). Risk for serious injury was not affected by helmet use (OR = 0.9). Risk of neck injury was increased in those struck by motor vehicles (OR = 4.0), hospitalized for any injury (OR = 2.0), and those who died (OR = 15.1), but neck injury was not affected by helmet use. CONCLUSIONS: Prevention of serious bicycle injuries cannot be accomplished through helmet use alone, and may require separation of cyclists from motor vehicles, and delaying cycling until children are developmentally ready. PMID- 9213157 TI - Characteristics and outcomes of self inflicted pediatric injuries: the role of method of suicide attempt. AB - OBJECTIVE: To examine the epidemiologic characteristics and clinical outcomes of self inflicted pediatric injuries in relation to the method of suicide attempt. METHODS: Using data from the National Pediatric Trauma Registry Phase II, a comparative analysis was conducted for children under 15 years of age who were admitted from 1 October 1988 through 30 April 1996 because of self inflicted injury by firearm (n = 28), hanging (n = 38), or jumping from heights (n = 21). RESULTS: Of the 87 cases under study, 90% occurred at home, and 86% occurred between noon and midnight, with a peak in early evening (between 6 pm and 7 pm) More than one quarter (29%) had preexisting mental disorders, such as disturbance of conduct and depression. Toxicological tests were conducted on admission on 40 (46%) of the patients; 20% tested positive for alcohol or other illicit drugs. The method of suicide attempt was associated with gender and age of the patients: 75% of the firearm cases and 82% of the hanging cases were boys compared with 29% of the jumping cases (p < 0.01); 79% of the hanging cases were aged 13 years or younger compared with 39% of the firearm cases and 48% of the jumping cases (p < 0.01). The mean injury severity score was 18.6 for the firearm cases and 16.3 for the hanging cases, significantly greater than 8.5 for the jumping cases (p < 0.02). Reflecting the differences in injury severity, firearm cases and hanging cases were more likely than jumping cases to be sent to intensive care units or operating rooms from emergency departments, and to develop complications during hospitalization. The case fatality rate was 50% for the firearm cases, 32% for the hanging cases, and 5% for the jumping cases (p < 0.01). On average, these patients stayed in hospitals for 11 days and 52% of those who were alive at discharge had at least one impairment in communication, cognition, or self care functions. CONCLUSION: Boys and older children tend to use more lethal methods in suicide attempts. Even in this age group, suicide attempts often involve psychiatric disorders and acute abuse of alcohol or other illicit drugs. Firearms are associated with significantly increased risk of inhospital fatality. The clinical outcomes of self inflicted injuries appear to be worse than other injuries treated in the same trauma centers. PMID- 9213158 TI - A seven item scale for the assessment of disabilities after child and adolescent injuries. AB - OBJECTIVES: To develop a scale to assess physical disabilities after child or adolescent injuries. SETTING: The three main hospitals of Jerusalem. METHODS: Telephone interviews pertaining to the injury's effect on the functioning of children 4-17 years old (n = 281) were carried out six months after an injury. Disabilities were recalled by the parents for the period immediately after the injury (short term) and at the time of interview (long term). Of 25 questions derived from the International Classification of Impairments, Disabilities and Handicaps, seven were selected: limitations in walking, running, getting up/lying down, moving in bed, going to the toilet, bathing/keeping personal hygiene, and dressing. Construct validity was tested using the usual, sport, school, and leisure time activities as the gold standard. RESULTS: The prevalence of short term disabilities ranged from 23.8% to 37.7% and of long term disabilities from 0.4% to 11.8%. Cronbach's alpha was 0.91 for the short term scale and over 0.90 for the different categories of the sociodemographic variables. It decreased to 0.66 for the long term scale. Sensitivity of the short term scale ranged from 77% to 89%, but was lower for the long term scale. Specificity varied from 72% to 84% and increased to 88% to 90%, six months after the injury. CONCLUSIONS: This scale could be used to study disability after injury among children and adolescents in different cultures. It is a simple method that does not require expert personnel and has relatively high validity and internal reliability. PMID- 9213159 TI - Program evaluation for prevention projects. PMID- 9213161 TI - Personality characteristics of the child accident repeater. 1967. PMID- 9213160 TI - Understanding suicide among indigenous adolescents: a review using the PRECEDE model. AB - AIM: To use the available literature to identify the causes of suicide among indigenous adolescents. METHOD: The PRECEDE model provided a framework to organize the material and identify the areas where relatively little research had been reported. RESULTS: The epidemiological diagnosis showed that suicide was greater in indigenous than non-indigenous populations and particularly high among adolescent males. Environments of native persons are characterized by remoteness, poverty, cultural displacement, and family disintegration. The educational and organizational diagnosis identified predisposing factors reflecting the social environments previously identified, the enabling factors of televised suicides, and firearm and alcohol availability, in conjunction with an absence of positive expectations. Finally the administrative and policy diagnosis identified a piecemeal, short term perspective, often lacking cultural sensitivity. Although there was more literature from the United States than from Canada, Australia or New Zealand, the pictures emerging were consistent, with problems being identified across continents. Literature was more abundant in relation to the epidemiological, environmental, and educational/ organizational diagnoses than in relation to policy and administration. CONCLUSION: The increased suicide rates among indigenous adolescents were not a product of their native origins, but of the social milieu in which these people generally found themselves. PMID- 9213162 TI - Behavioral antecedents of accidental injuries in early childhood: a study of twins. 1971. PMID- 9213163 TI - The Connecticut Childhood Injury Prevention Center--the first six years. PMID- 9213164 TI - A novel therapy for corticosteroid-dependent or corticosteroid-resistant patients with ulcerative colitis. PMID- 9213165 TI - Divergent effects of octreotide on glucose tolerance in patients with acromegaly. PMID- 9213166 TI - Inappropriate hemoglobin A1c level, interference or bonus? PMID- 9213167 TI - A pilot study of centrifugal leukocyte apheresis for corticosteroid-resistant active ulcerative colitis. AB - Corticosteroids are effective in bringing about a clinical remission in patients with ulcerative colitis. However, in severely relapsed cases, corticosteroids are not always effective even when a high dosage is administered. In addition, the long-term use of corticosteroids often causes serious side effects. Therefore, an alternative treatment for active ulcerative colitis is necessary in order to avoid these clinical problems. In the present pilot study, the efficacy of leukocytapheresis using a centrifugal procedure was evaluated for corticosteroid resistant, active ulcerative colitis. Fourteen patients with corticosteroid resistant severely active ulcerative colitis were treated by leukocytapheresis. Thirteen patients (92.9%) achieved clinical remission within 4 weeks after the apheresis, and remained in remission for 8 months on average without any additional corticosteroid therapy. In the remaining patient, in whom remission was not induced, a total colectomy was performed immediately after the fourth course of leukocytapheresis. No significant side effects were noticed throughout the therapy. Both colonoscopic and histological examinations confirmed the beneficial effect of this procedure in terms of the reduction of severe inflammation of the affected colon. We found that the expression of two adhesion molecules, L-selectin and VLA4a, on the surface of peripheral leukocytes was decreased after this new therapy. PMID- 9213168 TI - High incidence of thrombosis in Fabry's disease. AB - Although a high incidence of thrombotic accidents in Fabry's disease has been postulated, few investigations have been performed. To clarify the incidence of thrombosis in Fabry's disease, we undertook a systematic study on thrombosis in patients with Fabry's disease including hemizygous males and heterozygous females. Sixty patients with Fabry's disease (45 hemizygotes and 15 heterozygotes) from 36 Japanese families were subjected to clinical, biochemical and genetic investigations. We found that seven patients with Fabry's disease (4 hemizygous males and 3 heterozygous females) had experienced thrombotic accidents. Six of these thrombotic patients developed brain infarctions, including one man who had the complication of recurrent thrombophlebitis. The remaining woman showed central retinal artery occlusion and thrombophlebitis. We demonstrated a high incidence of thrombosis in Fabry's disease. Thrombotic accidents occurred not only in hemizygous males but also in heterozygous females. The complication of thrombotic accidents should be taken into account in patients with Fabry's disease. PMID- 9213169 TI - Problems in the initial diagnosis of renal infarction. AB - We retrospectively analyzed 20 cases of renal infarction to identify the problems in tentatively diagnosing renal infarction. The subjects consisted of 12 outpatients and 8 inpatients whose diagnosis was confirmed by renal scintigram and/or contrast computed tomography. Renal infarction was tentatively diagnosed in only 4 of the 12 outpatients. Causes of hospitalization were cerebral emboli in 5 cases, peripheral emboli in the extremities in 2 cases and one case involved percutaneous transmitral commissurotomy. On initial urinalysis, 11 cases (55%) showed less than 2+ hematuria using dipsticks to test for occult blood. The mean lactic dehydrogenase value was as high as 2,096 IU while the mean aspartate aminotransferase and mean alanine aminotransferase were 83.1 IU and 78.6 IU. Abdominal ultrasonography revealed abnormalities in only one of 18 cases. In conclusion, since only a moderate degree of hematuria was seen in about half the cases and it was difficult to detect renal abnormalities by ultrasonography, a tentative diagnosis of renal infarction may be difficult in some cases. PMID- 9213170 TI - Facioscapulohumeral muscular dystrophy: clinical diversity and genetic abnormalities in Japanese patients. AB - We studied 71 Japanese individuals, 42 patients (30 familial and 12 sporadic) suspected to have facioscapulohumeral muscular dystrophy (FSHD) and 29 family members, clinically and genetically using the chromosome 4qter DNA marker p13E 11. Early onset FSHD was detected in 7 patients, tortuosity of retinal arterioles and hearing impairment in 3 patients, progressive respiratory failure in 3 patients and limb-girdle type muscular weakness in 6 patients. Thirty-six (85.7%) FSHD patients, 3 asymptomatic family members and 1 of 35 healthy volunteers showed EcoRI digestion fragments shorter than 28kb. New mutations were detected in 25% of the patients with shorter EcoRI fragment. The age of disease onset appeared younger with successive generations in 6 parent-child pairs in FSHD families. We confirmed the existence of phenotypic and genetic diversities in Japanese patients with FSHD. It is still difficult to explain the phenotypic diversity merely by the size of the abnormal EcoRI fragment detected with the p13E-11 probe. PMID- 9213171 TI - Patent ductus arteriosus with combined valvular disease at age 91. AB - This report describes a 91-year-old patient with patent ductus arteriosus (PDA) complicated by combined valvular disease (CVD) (aortic and mitral stenosis, and aortic, mitral, pulmonic and tricuspid regurgitation). This patient seems to be the oldest living female with PDA and CVD hitherto reported in the medical literature. The patient developed several bouts of congestive heart failure which were treated medically. She not only has survived without surgical management, but is still enjoying her life at age 91. The features of PDA in the elderly are reviewed. PMID- 9213174 TI - Diffuse pulmonary amyloidosis with monoclonal IgG-kappa gammopathy. AB - A 62-year-old female with diffuse pulmonary amyloidosis developed abnormal radiographic findings while under observation for hyperimmunoglobulinemia over a ten-year period. Serum immunoglobulin G (IgG) was elevated (4,620 mg/dl), and associated with monoclonal gammopathy (M protein) of the kappa type, but no evident abnormalities were apparent in bone marrow. Chest radiograph and computed tomography showed a diffuse reticulonodular shadow in the bilateral lung. Thoracoscopic lung biopsy specimen revealed depositions of amyloid in the bronchus and pulmonary vessel. We emphasize that diffuse pulmonary amyloidosis should be considered a possible diagnosis in the presence of monoclonal immunoglobulin. PMID- 9213172 TI - A patient with acromegaly who showed remarkable improvement of hyperglycemia after treatment with octreotide. AB - A case with diabetes mellitus associated with growth hormone (GH)-producing pituitary adenoma is described. A 56-year-old woman who had been treated for diabetes mellitus for 3 years, was admitted for the treatment of hyperglycemia. She showed a few acromegalic features and her plasma GH level was 146 +/- 16 ng/ml. After improvement of plasma glucose level by insulin injection, octreotide therapy (100 micrograms/8 hours) was started. Seven days after the initiation of octreotide therapy, the fasting plasma glucose level was almost normalized without insulin injection. After the octreotide treatment, urinary C-peptide excretion was significantly decreased and the plasma GH level became within normal range. In this case, octreotide appears to have improved the insulin sensitivity by reducing the plasma GH level. PMID- 9213173 TI - A diabetic case with hemoglobin J-Meerut and low HbA1C levels. AB - A diabetic patient with hemoglobin (Hb) J-Meerut and low HbA1C levels is reported. An automatic glycohemoglobin analyzer used for the determination of HbA1C revealed an abnormal peak of the peripheral blood obtained from a Japanese female with diabetes. She showed a lower HbA1C level (3.7%) than expected from her fasting plasma glucose (172 mg/dl). High performance liquid chromatography and isoelectric focusing indicated that her abnormal hemoglobin was Hb J-Meerut [alpha 120(H3)Ala-->Glu] and it accounted for 28.3% of the total hemoglobin. Abnormal hemoglobinemia should be considered when a major discrepancy between the levels of HbA1C and fasting plasma glucose is observed. PMID- 9213175 TI - Interstitial pneumonitis related to granulocyte colony-stimulating factor administration following chemotherapy for elderly patients with non-Hodgkin's lymphoma. AB - We treated three cases of interstitial pneumonitis (IP) in 26 elderly (> or = 65 years old) patients with non-Hodgkin's lymphoma (NHL) who received the same chemotherapeutic protocol including granulocyte colony-stimulating factor (G-CSF) administration. Fortunately, all three patients recovered from IP spontaneously by discontinuation of G-CSF alone or with administration of corticosteroid. Because the duration and extent of neutrophilia induced by G-CSF administration was not different between the cases complicated by IP and those without IP, underlying pulmonary damage is suggested to be more involved than neutrophil count in the development of IP. PMID- 9213176 TI - The first Japanese case of Hb Santa Ana, an unstable abnormal hemoglobin, identified rapidly by electrospray ionization mass spectrometry. AB - Two members of a family had chronic hemolytic anemia due to unstable hemoglobin. The abnormal beta-chain with a molecular weight of 16 u smaller than the normal beta-chain was found within 5 minutes by analysis using electrospray ionization mass spectrometry. The substitution of amino acid was also determined rapidly by this new strategy. The leucine at the 88th position of the normal beta-chain was substituted by proline in the hemoglobin as in Hb Santa Ana. This is the first report of a Japanese case of Hb Santa Ana; the clinical course was similar to that in the previously reported cases. PMID- 9213177 TI - Aplastic anemia complicating Sjogren's syndrome. AB - A 47-year-old woman was referred to our hospital because of severe anemia and polyclonal gammopathy. She developed sicca syndrome after admission. Laboratory data revealed pancytopenia (white blood cells, 2,800/microliter; hemoglobin, 6.4 g/dl; platelets, 6.1 x 10(4)/microliter) and hyper gamma globulinemia (5.2 g/dl), and bone marrow was hypoplastic. Histology of the salivary gland showed infiltration of lymphocytes. We report a good response to immunosuppressive therapy in a rare case of aplastic anemia complicating Sjogren's syndrome. PMID- 9213179 TI - A case of non-insulin-dependent diabetes mellitus associated with anorexia nervosa. PMID- 9213178 TI - Progressive retinitis-encephalitis due to ganciclovir-resistant cytomegalovirus associated with aplastic anemia. AB - A 29-year-old woman with aplastic anemia who complained of visual disturbances and pain in the right eyeball was diagnosed as having cytomegalovirus (CMV) retinitis based upon the characteristic retinal changes and isolation of CMV. She received treatment with ganciclovir (GCV), and the retinitis initially responded for several months. However, the patient was found to have CMV lesions in the left eye followed by neurological symptoms. The CMV isolated just before her death was approximately 20 times more resistant to GCV than that isolated previously, suggesting that the GCV-resistant CMV had developed during the long term treatment with GCV. PMID- 9213180 TI - Pulmonary manifestations in von Recklinghausen's disease. PMID- 9213181 TI - Vertebral osteomyelitis in diabetes mellitus. PMID- 9213182 TI - Risk factors and incidence of coronary artery lesions in patients with abdominal aortic aneurysms. AB - We determined the incidence of coronary lesions by coronary angiography and the associated risk factors in 102 patients with abdominal aortic aneurysms (AAA). Old myocardial infarction was observed in 33 patients and angina pectoris in 27 patients. Coronary angiography revealed significant stenosis (> or = 75%) in 66 patients, including single-vessel disease in 22 patients and multiple-vessel disease in 44 patients. Angiography detected 21 coronary artery ectasia lesions, consisting of 10 saccular and 11 fusiform aneurysms in 18 patients. Significant coronary stenosis was present in approximately two-thirds of patients with AAA, suggesting that coronary angiography is a useful preoperative examination in patients with AAA. The incidence of coronary ectasia was also high in patients with this disease, perhaps because AAA and coronary ectasia involve similar pathogenic processes. There was considerable overlap in risk factors in patients with AAA and patients with coronary artery disease without AAA. However, advanced age and hypertension were more strongly associated with AAA than with coronary artery disease. PMID- 9213183 TI - Elevated serum lipoprotein (a) levels associated with ulcerative colitis in a young Japanese patient. AB - Thromboembolism has been shown to play a role in the pathogenesis of inflammatory bowel disease (IBD). A possibility exists that lipoprotein (a) [Lp(a)], a newly discovered prothrombotic factor, also participates in the development of at least some cases of IBD. Marked elevation of serum Lp(a) levels was observed in a young patient with ulcerative colitis. A biopsy specimen of the rectal mucosa showed findings compatible with ulcerative colitis, as well as small vessel thrombus occurring within the muscularis mucosa in the rectum. Serum Lp(a) levels were markedly elevated on admission (71 mg/dl), with a gradual decrease to 46 mg/dl on discharge. Moreover, serum Lp(a) levels decreased in parallel with clinical improvement. In the quiescent clinical stage, no small vessel thrombus was observed in the mucosa on follow-up colonoscopy. The association between IBD and hyper-Lp(a)-emia would be presumable but it has been, to our knowledge, previously unreported. The case reported here would be the first young patient, suggesting the presence of hyper-Lp(a)-emia and small vessel thrombus formation occurring in association with the development of ulcerative colitis. PMID- 9213184 TI - Gastrointestinal stromal tumor of the stomach. AB - We report a case of gastrointestinal stromal tumor (GIST) of the stomach. The patient was a 79-year-old woman with two gastric submucosal tumors detected by ultrasonography. Proximal gastrectomy was carried out and the tumors were diagnosed as GIST by histological and immunohistochemical investigations. Mesenchymal tumors of the gastrointestinal tract have been traditionally regarded as largely leiomyomatous lesions. However, GIST and other tumors have been distinguished recently on the basis of tumor cell differentiation shown by immunohistochemical studies. We discuss the concept and the immunohistochemical characteristics of GIST. PMID- 9213185 TI - A functioning black adenoma of the adrenal gland. AB - A 53-year-old female had clinical and laboratory findings suggestive of Cushing's syndrome. In contrast to the Cushing's syndrome caused by cortical adenoma, a high level of urinary 17-ketosteroids (17-KS) was also noted. Imaging studies revealed a right adrenal tumor. Right adrenectomy was performed; the surgical specimen revealed a black adenoma consisting of compact cells with numerous pigments which seemed to be lipofuscin in nature. The present case indicates that black adenoma as well as adrenocortical carcinoma should be suspected, when patients with Cushing's syndrome show an increased level of urinary 17-KS excretion. PMID- 9213186 TI - Pheochromocytoma in a long-term hemodialysis patient, discovered as an adrenal incidentaloma. AB - A case of pheochromocytoma was discovered incidentally during long-term hemodialysis for chronic renal failure due to acquired cystic kidney disease. A 52-year-old male patient was examined for weight loss of 3 kg during over a period of 3 months. Abdominal computed tomography (CT) revealed a left adrenal mass (3.0 cm in size). Plasma adrenaline and noradrenaline were increased to 521 pg/ml and 1,341 pg/ml, respectively, and the metoclopramide provocative test was positive. Further, in the scintiscan using 123I-metaiodobenzylguanidine (MIBG), an accumulation of the radionuclide in the left adrenal tumor region was confirmed. The patient is currently under observation and conservative treatment due to the possible occurrence of arterial hypotension after resection of the tumor and to lesser possibility of the malignancy. PMID- 9213187 TI - Adult-onset type II citrullinemia: clinical pictures before and after liver transplantation. AB - In a 25-year-old man with adult-onset type II citrullinemia, liver transplantation resulted in elimination of hyperammonemia and amino acid abnormalities associated with the disease. Postoperatively, a high intensity area in the right cingulate gyrus on a T2-weighted brain magnetic resonance imaging (MRI) also disappeared, suggesting that it reflected an early reversible lesion due to the hyperammonemia. Moreover, the serum level of pancreatic secretory trypsin inhibitor (PSTI), which had been elevated, was normalized. Since the levels of PSTI mRNA and PSTI have been reported to be increased in the livers of type II citrullinemia patients, measurement of serum PSTI levels could aid in the diagnosis of this disease. PMID- 9213188 TI - Distal acinar emphysema and interstitial pneumonia in a patient with von Recklinghausen's disease: five-year observation following quitting smoking. AB - Cystic lesion, malignancy and interstitial pneumonia are well-known as pulmonary complications of patients with von Recklinghausen's disease. We report herein an unusual patient with distal acinar emphysema and interstitial pneumonia of prominent hypercellularity demonstrated by transbronchial biopsy and broncho alveolar lavage fluid (BALF). Six months after quitting smoking, the total cell count of BALF was remarkably reduced. This patient remains stable under 5-year observation in terms of symptoms and findings of both BALF and pulmonary function tests. Quitting smoking may have facilitated a favorable prognosis for the particular lung disease complicated in this patient. PMID- 9213189 TI - Primary lymphoma of the heart, diagnosed antemortem. AB - Primary cardiac lymphomas diagnosed antemortem are extremely rare. We present a case of primary cardiac lymphoma initially diagnosed antemortem by cytologic examination of pericardial effusion fluid. Echocardiography suggested the presence of a tumor localized at the right ventricular free wall. The cytologic examination of pericardial effusion was effective in establishing the correct antemortem diagnosis. PMID- 9213190 TI - Non-Hodgkin's lymphoma with pulmonary infiltrates mimicking miliary tuberculosis. AB - Miliary infiltrates observed on chest films in non-Hodgkin's lymphoma are extremely rare. We report a case with pulmonary infiltrates mimicking miliary tuberculosis associated with prominent eosinophilia and elevated IgE levels. The levels of circulating eosinophils correlated with disease activity as they transiently returned to normal after effective chemotherapy in a short period. However, the patient developed acute respiratory failure due to the rapid progression of the disease even with intensive chemotherapy. We emphasize that small nodular shadows appear to be a sign of the rapid progression of the disease and a poor prognosis. PMID- 9213191 TI - Peripheral primitive neuroectodermal tumor involving the paravertebral and retroperitoneal regions. AB - A rare case of peripheral primitive neuroectodermal tumor (PNET) is reported. A 68-year-old woman complaining of lumbago was admitted to our hospital. Diagnosis was made based on pathological findings characterized by Homer Wright-type rosettes. Ultrastructural examination showed the presence of neurosecretory granules and short cytoplasmic processes, which were highly suggestive of neural differentiation. Chromosomal analysis of the neoplastic cells revealed translocation (11;22)(q24;q12), which is often found in Ewing's sarcoma and Askin tumor. These results strengthen the hypothesis of a common histogenesis for these small round cell tumors, and suggest common oncogenesis for these neoplasms. PMID- 9213192 TI - Diffuse alveolar hemorrhage associated with proteinase 3-specific anti-neutrophil cytoplasmic antibodies. AB - A 31-year-old man was referred to our hospital for the management of progressive diffuse alveolar hemorrhage associated with renal dysfunction. Leukocytoclastic vasculitis was shown by skin biopsy and crescentic glomerulonephritis was also detected, in addition to positivity for proteinase 3-specific anti-neutrophil cytoplasmic antibodies (PR3-ANCA). The patient was diagnosed as a rare case of PR3-ANCA-positive pulmonary-renal vasculitic syndrome without granulomatous lesions. There was a good response to combination therapy with steroids and cyclophosphamide. PMID- 9213193 TI - Association of progressive systemic sclerosis with pulmonary sarcoidosis. Just a chance occurrence? AB - A 46-year-old Japanese woman has been followed up for 3 years due to interstitial pneumonia associated with progressive systemic sclerosis (PSS). During this follow-up period, chest roentgenogram revealed additional diffuse nodular shadows. She was diagnosed as having pulmonary sarcoidosis, which was confirmed by the presence of epitheloid granulomas within the alveolar septa. She was successfully treated with corticosteroids and recovered almost completely without worsening pulmonary abnormalities caused by her PSS. The independent clinical courses of these two diseases in the present case suggest that the complication of PSS and sarcoidosis in this patient may be coincidental. PMID- 9213194 TI - Cryofibrinogenemia with polyarthralgia, Raynaud's phenomenon and acral ulcer in a patient with Graves' disease treated with methimazole. AB - Cryofibrinogenemia is a cryopathy in which hypersensitivity to cold is a prominent feature. Cryofibrinogenemia developed in an 18-year-old Japanese female patient during methimazole therapy for Graves' disease. She developed cryopathy (livedo reticularis, Raynaud's phenomenon and acral ulcer) and polyarthralgia during methimazole therapy, and we detected cryofibrinogen in her plasma. Her symptoms resolved after administration of prostaglandins and anticoagulants. Several reports indicate that methimazole therapy induces autoantibody-related disease. In the present case, we cannot exclude the possibility that methimazole therapy contributed to the cryofibrinogenemia. PMID- 9213195 TI - Discitis, infectious arthritis, and bacterial meningitis in a patient with pancreatic diabetes. AB - A 63-year-old woman with pancreatic diabetes after a total pancreatectomy and splenectomy developed discitis of the L2/3 intervertebral disk. Rapidly she also developed infectious arthritis of the left knee joint and bacterial meningitis. Aspirate from the left knee contained Enterococcus faecalis. The diagnosis of discitis is generally difficult in the initial period of disease, and patients with diabetes or splenectomy are susceptible to rapid progression of the infection. Early diagnosis of discitis using magnetic resonance imaging of the spine and treatment with antibiotics might have altered her clinical course. PMID- 9213196 TI - Levosimendan enhances left ventricular systolic and diastolic function in conscious dogs with pacing-induced cardiomyopathy. AB - We examined the left ventricular (LV) mechanical actions of levosimendan (LSM) before and after the development of pacing-induced cardiomyopathy in conscious dogs chronically instrumented for measurement of aortic and LV pressure, +dP/dt, subendocardial segment length, and cardiac output (CO). The slope (Mw) of the regional preload recruitable stroke work relation was used to assess myocardial contractility. Diastolic function was evaluated with a time constant of isovolumic relaxation (tau), the maximal rate of segment-lengthening velocity (dL/dt), and a regional chamber-stiffness constant (Kp). On different experimental days, dogs were assigned to receive LSM (12- or 24-microgram/kg loading dose and 0.2 or 0.4 microgram/kg/min infusion) before rapid ventricular pacing was initiated. Dogs were then paced at 240 beats/min for 22 +/- 2 days, and the low and high doses of LSM were repeated on separate days. LSM increased Mw and +dP/dt in dogs before the initiation of pacing, consistent with enhanced myocardial contractility. LSM also improved indices of LV diastolic function (decreases in tau and Kp and increases in dL/dt) in dogs before pacing. Rapid ventricular pacing over a 3-week period increased LV end-diastolic pressure and produced systolic (decreases in Mw and +dP/dt) and diastolic (increases in tau and Kp and decreases in dL/dt) dysfunction. LSM significantly (p < 0.05) increased Mw (54 +/- 3 to 98 +/- 6 mm Hg) +dP/dt and dL/dt (57 +/- 13 to 72 +/- 13 mm/s) and decreased tau (66 +/- 4 to 52 +/- 3 ms) and Kp (1.14 +/- 0.14 to 0.71 +/- 0.03 mm-1) in the presence of LV dysfunction. In contrast to the findings in normal dogs, however, LSM did not alter heart rate and calculated indices of myocardial oxygen consumption in dogs after pacing. The findings indicate that LSM produces favorable alterations in hemodynamics and positive inotropic and lusitropic effects in conscious dogs with left ventricular dysfunction. PMID- 9213197 TI - Combined effects of caffeine and nicotine on cardiovascular hemodynamics in canine model. AB - The independent effects of caffeine and nicotine on cardiodynamics are well documented, but combined effects of both are not reported. Initially, in phase I, 18 experiments were performed to study the dose-response curve of both the drugs. In phases II and III, 13 mongrel dogs were subjected to 30 experiments. In phase II, caffeine, 5 mg/kg, was given i.v. followed by nicotine, 50 micrograms/kg, and in phase III, the sequence of drug administration was reversed to study the effects on hemodynamics. In phase II, caffeine did not show significant changes in all the cardiovascular parameters, but nicotine administration after caffeine produced marked significant synergistic excitatory effects: the rate of increase of the first derivative of left ventricular pressure (dP/dt) increased from 1,101 +/- 111 to 3,194 +/- 872 (p < 0.003). In phase III, nicotine significantly increased heart rate, mean arterial pressures; left ventricular end-diastolic pressure (LVEDP); and pulmonary artery, pulmonary capillary wedge, and right atrial pressures. Nicotine increased dP/dt (964 +/- 182 to 1,639 +/- 60 mm Hg/s; p < 0.004). The excitatory effects of nicotine were attenuated by administration of caffeine (dP/dt, 918 +/- 140 reduced to 715 +/- 144 mm Hg/s; p < 0.04). Caffeine and nicotine, alone, caused nonsignificant and significant increases in hemodynamics, respectively. In combination, caffeine + nicotine administration produced significant synergistic excitatory effects in dogs. On the other hand, the nicotine + caffeine combination caused attenuation by caffeine of the excitatory effects produced by nicotine. PMID- 9213198 TI - Effect of poloxamer 407 on the activity of microsomal 3-hydroxy-3-methylglutaryl CoA reductase in rats. AB - A single 300-mg i.p. injection of poloxamer 407 (P-407, also called Pluronic F 127) in rats produces a marked hypercholesterolemia for a minimum of 96 h. The purpose of this investigation was to determine mechanisms by which P-407 causes hypercholesterolemia. The enzyme targeted for investigation is the rate-limiting enzyme in cholesterolgenesis, 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. Injection of P-407 in fasted rats resulted in a significant (p < 0.05) elevation in plasma cholesterol (61.2 +/- 4.2 mg/dl) as soon as 1 h after injection compared with sham-injected controls (50.1 +/- 3.7 mg/dl). Plasma cholesterol (CHO) 24 h after injection of P-407 was 449 +/- 57 mg/dl, with the fastest rate of accumulation occurring from 1 to 12 h (approximately 16.6 mg/dl/h). Over the concentration range of 0-5 mM, P-407 did not appear significantly to affect the activity of HMG-CoA reductase in vitro. However, the enzymatic activity assayed in microsomal fractions isolated from the livers of P 407-injected rats reached a maximum of 262 +/- 42.6 pmol mevalonate/min/mg approximately 15 h after injection, with a subsequent decline to control activity (94.1 +/- 8.7 pmol/min/mg) at approximately 40 h after injection. At 48 h after injection of P-407, the activity of HMG-CoA reductase significantly (p < 0.05) decreased below control values with a mean activity of 9.4 +/- 1.2 pmol/min/mg. The CHO concentrations in hepatic tissue were significantly (p < 0.01) increased at 2 h (3.26 +/- 0.19 mg/g) and 4 h (3.75 +/- 0.38 mg/g) and significantly (p < .01) reduced at 15 h (1.56 +/- 0.19 mg/g) after injection of P-407 compared with tissue CHO concentrations determined in control animals (2.65 +/- 0.18 mg/g). However, the hepatic CHO content appeared to return to control values by 24 h (mean +/- SEM, 2.61 +/- 0.08 mg/g) after injection. These data suggest that the activity of HMG-CoA reductase is regulated by some indirect mechanism(s) after injection of P-407 in rats. PMID- 9213199 TI - Cardioprotective and hypolipidemic effects of nisoldipine in the JCR:LA-cp rat. AB - The JCR:LA-cp rat exhibits the obesity/insulin resistance/hypertriglyceridemia syndrome in an extreme form. These normotensive rats spontaneously develop advanced atherosclerosis and ischemic myocardial lesions. The calcium channel antagonist, nisoldipine, was administered to obese rats of the JCR:LA-cp strain in drinking water at a dose of 1 mg/kg from age 6 weeks. Nisoldipine-treated rats showed no change in food consumption or body weight compared with control animals. Plasma glucose and insulin levels also were unchanged in the nisoldipine treated rats. Insulin-mediated total glucose turnover, an index of insulin sensitivity as measured by euglycemic insulin clamp, was similarly not improved. Serum triglyceride levels in obese male rats were markedly reduced (57%; p < 0.001, at age 12 weeks), whereas obese female rats showed no significant change in triglyceride levels and an increase in esterified cholesterol in response to nisoldipine treatment. The impaired endothelium-dependent (nitric oxide-mediated) vascular relaxation of the male cp/cp rats was not improved by nisoldipine treatment. The severity of atherosclerotic raised lesions in the aortic arch of male cp/cp rats was significantly reduced (p < 0.01) by nisoldipine treatment, and this was accompanied by a major reduction in the incidence of ischemic myocardial lesions (85%; p < 0.01). Thus nisoldipine treatment ameliorates atherosclerotic damage and myocardial injury even in the presence of gross obesity, hyperinsulinemia, and significant hyperlipidemia. This effect appears to involve protection of the vascular wall from atherogenesis and probably antivasocontractile effects at the smooth muscle level as well. PMID- 9213200 TI - Effects of tilisolol, a nonselective beta-adrenergic blocker, on the membrane currents of isolated guinea pig ventricular myocytes. AB - The effects of tilisolol, a nonselective beta-adrenoceptor blocker, on transmembrane ionic currents were studied in single guinea pig ventricular myocytes by using the whole-cell voltage clamp technique. In the absence of beta adrenergic stimulation, 10 microM tilisolol, a concentration higher than that used in the clinical therapeutic regimen, did not affect the L-type Ca2+ current (ICa), the inwardly rectifying K+ current (IK1), or the delayed rectifying K+ current (IK). In addition, it did not induce currents through the adenosine triphosphate (ATP)-sensitive K+ channels. However, under the nonselective beta adrenergic stimulation with 1 microM isoproterenol, 1 microM tilisolol almost completely reversed the agonist-induced increase of IK. The increase of ICa by isoproterenol was blocked only by approximately 30% with tilisolol. We concluded that, at therapeutic concentrations (0.01-0.15 microM), tilisolol is a pure beta adrenoceptor antagonist that has no direct effects on the transmembrane ionic currents of mammalian ventricular myocytes, such as ICa, IK1, or IK. Comparison of the dose-dependent effects of tilisolol on ICa and IK suggested that tilisolol may selectively inhibit catecholamine-induced increase of IK at the therapeutic concentrations. The virtually selective inhibition of IK, leaving ICa intact, may be favorable to prevent the catecholamine-induced arrhythmia without inhibiting contraction. PMID- 9213201 TI - The increase in blood pressure induced by inhibition of nitric oxide synthase in anesthetized Wistar rats is inversely related to basal blood pressure value. AB - Nitric oxide produced by endothelial cells is a mediator involved in the regulation of vascular tone. Indeed, in vitro inhibition of basal nitric oxide release increase responses to vasoconstrictor agents, and in vivo, the parenteral or dietary administration of nitric oxide inhibitors produces an increase in blood pressure. However, the correlation of nitric oxide production and basal blood pressure in normotensive subjects is still unclear. In this study, we showed that administration, in urethane-anesthetized Wistar rat, of two inhibitors of nitric oxide synthase, such as NG-nitro-L-arginine methyl ester and methylguanidine, produced a significant increase in mean arterial blood pressure that was inversely related to basal mean arterial blood pressure at all doses tested. On the other hand, the increase in mean arterial blood pressure induced by infusion of angiotensin II did not correlate with basal mean arterial blood pressure. The pretreatment of rats with hexamethonium did not change the results observed in normal rats, ruling out an involvement of sympathetic nervous system. In conclusion, this study further confirms the presence of a tonic amount of nitric oxide, produced by endothelial cells in the bloodstream, which plays a key role in the regulation of basal blood pressure, and its reduction or inhibition may be the main cause of certain pathologic conditions characterized by high levels of blood pressure. PMID- 9213202 TI - Angiotensin II-induced phosphoinositide production and atrial natriuretic peptide release in rat atrial tissue. AB - The effect of angiotensin II (Ang II) on inositol phosphate (IP) production and atrial natriuretic peptide (ANP) release was studied in sliced rat atrial tissue. The ability of Ang II (10(-7) M) to stimulate IP accumulation was detected after 1 min of incubation, and the maximal increase was observed at 5 min. In (2-3H) inositol-labeled atrial tissue, Ang II induced the formation of (2-3H) inositol monophosphate (IP1) in a dose-dependent manner. The effect of Ang II (10(-7) M) on IP1 was prevented by losartan (10(-7) M) but was not affected by PD123319 (10( 7) M). Similar effects were observed on Ang II-induced ANP release in the presence of these antagonists. The mechanism of ANP liberation induced by this peptide was independent of cyclic adenosine monophosphate (cAMP) and regulated by nitric oxide (NO). The role of Ca2+ in the effect of Ang II was tested by 1,2-bis (o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid tetra (acetoxymethyl) ester (BAPTA-AM; 10(-5) M), a chelator of intracellular Ca2+ that prevented the release of ANP by Ang II stimulation. We concluded that Ang II induced IP production and ANP release through AT1 receptors. Stimulation of ANP release by Ang II was dependent on intracellular Ca2+. PMID- 9213203 TI - Assessment of antihypertensive effect by blood pressure monitoring: applications to bisoprolol and lisinopril in a double-blind study. AB - The aim of this study was to evaluate the antihypertensive effect of drugs according to the initial ambulatory blood pressure (BP) level. After a 15-day placebo run-in period, 105 patients with moderate essential hypertension (mean age, 52 years) underwent 24-h BP monitoring (spacelabs: 1 measure/15 min). Patients were subdivided into two groups: the "High" group, with 24-h mean values of systolic BP (SBP) > 137 or diastolic BP (DBP) > 87 mm Hg, and the "Low" group, with SBP < or = 137 and DBP < or = 87 mm Hg. All patients received, in a random and double-blind design, either bisoprolol (10 mg q.d.) or lisinopril (20 mg q.d.) for 8 weeks. At the end of this active treatment period, office and ambulatory BP measurements were performed. Casual measurements revealed similar BP decreases in all subgroups receiving bisoprolol and lisinopril; BP monitoring showed that the antihypertensive effect depended on the baseline mean 24-h value; -15/-12 mm Hg for bisoprolol and -18/-13 mm Hg for lisinopril in the High group; 7/-6 mm Hg for bisoprolol and -6/-6 mm Hg for lisinopril in the Low group. This study shows that the antihypertensive effect depended on initial ambulatory BP values, with a lower BP decrease in the Low group. Assessment of the antihypertensive effect on ambulatory BP is useful in clinical trials. PMID- 9213204 TI - Hemodynamic and autonomic effects of intravenous saterinone in patients with chronic heart failure. AB - In this study, the hemodynamic and neurohumoral/autonomic effects of intravenous saterinone (a selective phosphodiesterase type III inhibitor, with additional alpha 1-blocking properties) were evaluated. In a double-blind, placebo controlled design, 36 patients with moderate to severe heart failure were studied (saterinone, n = 24; placebo, n = 12). Invasive hemodynamic measurements, by using right-heart catheterization, were performed, as well as measurement of plasma neurohormones and analysis of heart rate variability (HRV), to study drug influences on neurohumoral activation and autonomic tone. Systemic vascular resistance significantly decreased during saterinone infusion, accompanied by a decrease in systemic blood pressure (both p values < 0.05) and an increase in heart rate (p = 0.05). Filling pressures also decreased during saterinone, but this was statistically significant only for pulmonary capillary wedge pressure, whereas the cardiac index remained unaffected. Plasma neurohormones (norepinephrine, epinephrine, and renin activity) were not significantly influenced by saterinone. HRV analysis revealed no significant effect of saterinone on autonomic tone. These results suggest that intravenous saterinone has a significant vasodilating effect in patients with moderate to severe chronic heart failure (CHF), without exerting an adverse effect on the autonomic nervous system, as demonstrated by assessment of plasma neurohormones and HRV analysis. PMID- 9213205 TI - Inhibitory effects of efonidipine hydrochloride on contraction induced by several vasoconstrictors in porcine coronary artery: comparison with effects of nifedipine and nisoldipine. AB - We studied the effects of efonidipine hydrochloride (efonidipine), a 1,4 dihydropyridine derivative, on contractions induced by high-K+ solution (high K+), serotonin (5-HT), U46619, which is a stable analog of thromboxane A2, and endothelin-1 (ET-1) in comparison with those of nifedipine and nisoldipine in porcine coronary arteries. The effects of the drugs were compared after 1- and 3 h incubations. Efonidipine, nifedipine, and nisoldipine each inhibited the contractions induced by these vasoconstrictors. The inhibition of high-K(+)- and 5-HT-induced contractions by efonidipine, but not by nifedipine and nisoldipine, increased when the incubation time was prolonged, whereas the inhibition of U46619- and ET-1-induced contractions was not altered. The potency of efonidipine on U46619- and ET-1-induced contractions was greater than that of nifedipine and equivalent to that of nisoldipine. Thus the inhibitory effect of efonidipine on U46619- and ET-1-induced contractions seems to be stronger than its effects on high-K(+)- or 5-HT-induced contractions, in contrast to the effects of other dihydropyridines. In an additional series of experiments, efonidipine did not inhibit U46619-induced contractions in Ca2(+)-free solution or in the presence of nifedipine. Moreover, efonidipine did not inhibit the specific binding of [3H]SQ 29,548, a thromboxane A2 antagonist, to porcine coronary arterial membrane. Therefore we think that the inhibitory effect of efonidipine on contractions induced by vasoconstrictors was caused by blockade of Ca2+ influx through L-type Ca2+ channels. However, some unknown mechanism(s) in addition to this effect on Ca2+ channels may contribute to the effect of efonidipine on U46619- and ET-1 induced contractions. PMID- 9213206 TI - Time course of delayed myocardial protection after transient adenosine A1 receptor activation in the rabbit. AB - Ischaemic preconditioning of myocardium enhances tolerance to infarction in a biphasic manner. Adenosine A1-receptor activation has been implicated as a trigger of both the early the late phases of protection in rabbit myocardium. Delayed protection against myocardial infarction in vivo was previously shown to occur 24 h after transient adenosine A1-receptor activation with the selective agonist 2-chloro-N6-cyclopentyladenosine (CCPA). Our studies examined the time course of CCPA-induced delayed myocardial protection and a possible mechanism of protection, the elevation of the cytoprotective inducible 72-kDa heat-shock protein (hsp70i). Rabbits were pretreated with a single dose of CCPA 100 micrograms/kg or saline i.v. Twenty-four, 48, 72, or 96 h after treatment, they were anaesthetised, and a left branch of the circumflex coronary artery was reversibly occluded for 30 min, followed by 120 min reperfusion. Infarct size was determined as a percentage of the myocardial risk volume by using triphenyltetrazolium staining. Approximately 50% reduction in infarct-to-risk volume ratio was observed 24, 48, and 72 h after CCPA pretreatment, compared with time-matched saline-pretreated controls. No infarct limitation was observed 96 h after CCPA pretreatment. Differences in infarct size were not related to differences in myocardial risk zone size or systemic haemodynamic parameters during the infarct protocol. Left ventricular tissue harvested from a separate cohort of animals pretreated with CCPA, 100 micrograms/kg, was assessed for content of hsp70i by Western blot analysis. The protein was not induced by CCPA treatment at any time point during the period in which cardioprotection was observed. We conclude that transient adenosine A1-receptor activation produces a delayed and prolonged period of enhanced resilience to ischaemia in rabbit myocardium. This is probably the result of an adaptive mechanism but does not involve elevation of hsp70i. PMID- 9213207 TI - Centrally administered calcium increases the maximum vagal activation of baroreceptor reflex control of heart rate in spontaneously hypertensive rats. AB - Baroreceptor reflex control of heart rate (HR) was examined before and 15 min after intracerebroventricular infusion (i.c.v.) of 10 microliters of high-Ca2+ solution (Ca2+, 16.3 mM) in conscious spontaneously hypertensive (SHR) and Wistar Kyoto rats (WKY). The slope of the individual regression line of the relation between reflex HR changes (delta HR) and changes in mean arterial pressure (delta MAP) produced by bolus injections of phenylephrine or sodium nitroprusside (delta HR/delta MAP; beats/min/ mm Hg) for bradycardia was significantly less in SHR ( 0.60 +/- 0.30; n = 10) than in WKY (-1.78 +/- 0.27; n = 10; p < 0.01) at baseline. The slope increased in SHR during administration of high Ca2+ to -1.39 +/- 0.17 (p < 0.01) but not in WKY: In contrast, no significant changes were observed for the reflex tachycardia both in SHR (n = 7) and WKY (n = 10). Further, we analyzed sigmoidal MAP-HR reflex curves in SHR with i.c.v. of either high Ca2+ (n = 6) or artificial cerebrospinal fluid (aCSF; n = 5). Administration of high Ca2+ reduced the bradycardic plateau and increased HR range without changes in average gain. Our results suggest a modulatory role for central Ca2+ in the baroreceptor reflex control of HR in SHR. PMID- 9213208 TI - In vivo evidence of positive inotropism of EMD 57033 through calcium sensitization. AB - The previous separation of the racemic cardiotonic thiadiazinone derivative EMD 53998 yielded two enantiomers with different pharmacologic properties: EMD 57,033, a potent Ca2+ sensitizer with some residual phosphodiesterase III (PDE III) inhibition, and EMD 57,439, a pure PDE III inhibitor. Although numerous in vitro studies demonstrated the ability of EMD 57,033 to increase the responsiveness of cardiac contractile proteins to Ca2+, in vivo evidence for such an action is lacking. Because there is no possibility of directly proving Ca2+ sensitization in vivo, we attempted to exclude PDE III inhibition as a major contributing component of the positive inotropic action of EMD 57,033. In anesthetized rats, EMD 57,033 increased left ventricular (LV) first derivative of change in systolic pressure over time (dP/dt max) without affecting blood pressure. In contrast, the PDE III-inhibitory enantiomer EMD 57,439 decreased blood pressure. The pattern of hemodynamic effects in anesthetized dogs revealed similar differences between EMD 57,033 and PDE inhibitors. Thus the increase in LV dP/dt max in response to EMD 57,033 was not accompanied by changes of heart rate and blood pressure. As expected for PDE inhibitors, pimobendan and milrinone increased cardiac contractile force in dogs, concomitant, however, with tachycardia, hypotension, and a decrease in total peripheral resistance. When regional contractility was measured separately in two different areas of the dog myocardium, the positive inotropic action of the PDE inhibitors pimobendan and milrinone was antagonized by local coronary infusion of acetylcholine. The cardiotonic effect of the Ca2+ sensitizer EMD 57,033 was entirely resistant to inhibition by acetylcholine. In conscious dogs, beta-blockade markedly attenuated the increase in LV dP/dt max produced by two different doses of the PDE III inhibitor EMD 57,439. In contrast, a dose of EMD 57,033 equieffective in positive inotropic action with the lower dose of EMD 57,439 remained unaffected by < b tau blockade. We concluded (a) that EMD 57,033 increases cardiac contractile force in two species in vivo, (b) that this action is independent of the cardiac cyclic adenosine monophosphate (AMP) system, (c) that EMD 57,033 does not reduce blood pressure and increase heart rate, an action indicative of PDE inhibition, and (d) that, on the basis of numerous previous in vitro findings, the mechanism of action of EMD 57,033, also in vivo, is consistent with sensitization of the cardiac myofibrils to Ca2+. Of special importance is the finding that this Ca2+ sensitizer at appropriate doses may be able to improve systolic function without adverse effects on diastolic function, as indicated by a slight decrease in left ventricular end-diastolic pressure. PMID- 9213209 TI - Prejunctional neuropeptide Y receptors in human kidney and atrium. AB - The aim of our study was to characterize functionally prejunctional neuropeptide Y (NPY) receptors in human and rabbit renal cortex, as well as in human right atrium. Segments of human atrial appendages and of human and rabbit renal cortex were preincubated with [3H]noradrenaline, superfused with Krebs-Henseleit solution and stimulated electrically in superfusion chambers. The stimulation induced outflow of radioactivity was taken as an index of endogenous noradrenaline release. The effects of subtype-selective NPY analogs on the stimulation-induced noradrenaline release were studied. NPY, its endogenous analog, peptide YY, and its C-terminal fragment, NPY13-36, but not its analog, [Leu31,Pro34]NPY, concentration dependently (1-100 nM) inhibited [3H]noradrenaline release in all tissues studied. NPY-induced inhibition of [3H]noradrenaline release in human and rabbit kidney was abolished by pretreatment with pertussis toxin. We conclude that prejunctional inhibition of noradrenaline release in human heart and human and rabbit kidney occurs through NPY receptors of the Y2 subtype, which appear to couple to a pertussis toxin sensitive G protein. PMID- 9213210 TI - Zatebradine slows ectopic ventricular rhythms in canine heart 24 hours after coronary artery ligation. AB - Arrhythmias occur 24 h after occlusion of the left anterior descending (LAD) coronary artery in the canine heart and have been attributed to the abnormal spontaneous activity in subendocardial Purkinje fibers, which are markedly depolarized. The major current underlying normal automaticity in these fibers is i(f). Although the i(f) activation range is generally considered to be more negative than the diastolic membrane potential in these depolarized fibers in infarcts, this activation range has been shown to shift in a positive direction in response to hormonal influences. Thus i(f) could still mediate automaticity in these fibers in infarcts. Furthermore, recent reports indicate that a depolarizing diastolic current, probably i(f), also can be measured in ventricular muscle during abnormal experimental conditions, which may occur during ischemia. To test whether there is a role of i(f) currents in sustaining ventricular ectopy, we administered the selective i(f) channel blocker, zatebradine, 24 h after LAD ligation in canine hearts. We report that intravenous injections of zatebradine (0.25 or 1.0 mg/kg) significantly slow ventricular rhythms (with average reductions of 19 or 26%, respectively). Moreover, because zatebradine also slows sinus nodal rate, it can lead to an increased incidence of ectopic beats. However, during right atrial pacing, when sinus slowing has no effect on ventricular rhythms, capture of ventricular rhythms occurs at lower rates in the presence of zatebradine. The reduction of capture threshold is comparable to the reduction in the rate of the ectopic rhythm. Thus zatebradine eliminated the arrhythmia when the right atrium was paced at the original sinus rate. PMID- 9213211 TI - Effects of 5-HT4-receptor agonists, cisapride, mosapride citrate, and zacopride, on cardiac action potentials in guinea pig isolated papillary muscles. AB - The purpose of this study was to examine the effects of 5-HT4-receptor agonists cisapride, mosapride citrate (mosapride), and zacopride on action potentials (APs) in guinea pig isolated papillary muscles. Cisapride (0.1-3 microM) concentration-relatedly prolonged the duration of APs (APD) without affecting the other AP parameters. Mosapride and its main metabolite M1 (des-4-fluoro-benzyl mosapride) did not affect APs at 10 microM. Zacopride at 10 microM shortened APD, and the APD-shortening effect was not affect by GR113808 (10 microM), a 5-HT4 receptor antagonist. The cisapride (1 microM)-induced prolongation of APD was not affected by GR113808 (10 microM), ritanserin (10 microM), a 5-HT2A/2C-receptor antagonist, or prazosin (10 microM), an alpha 1-receptor antagonist. The same concentrations of GR113808, ritanserin, and prazosin did not affect APD. Clofilium, a class III antiarrhythmic agent, prolonged APD; the effect was more pronounced at a stimulus frequency of 0.3 Hz than at 2.0 Hz. Cisapride did not exert such reverse use dependence, suggesting that its mechanism of action is different from that of clofilium. These results suggest that cisapride prolongs APD without involvement of 5-HT2, 5-HT4, or alpha 1 receptors. Mosapride is unlikely to induce the prolongation of electrocardiographic QT intervals correlated with the prolongation of APD in isolated ventricular muscles. PMID- 9213212 TI - Evidence of increased sympathetic vasomotor drive with shorter acting dihydropyridine calcium channel antagonists in human hypertension: a study using spectral analysis of RR interval and systolic arterial pressure variability. AB - We studied the effects of common antihypertensive regimens on autonomic cardiovascular control. We considered calcium channel antagonists (nicardipine twice a day and isradipine once a day, respectively) and also examined, as reference treatments, once-a-day atenolol and cilazapril. A noninvasive evaluation of autonomic cardiovascular profile was obtained with spectral analysis of RR interval and systolic arterial pressure variability. We studied moderate essential hypertensives before and after 2 weeks of treatment, both at rest and during active standing and mental arithmetic. All treatments reduced arterial pressure equally well; however, marked differences in spectral profiles were observed. The low-frequency spectral component of RR interval variability [in normalized units, marker of sympathetic modulation of the sinoatrial (SA) node] tended to be greater at rest and during stimuli (p < 0.001) in subjects treated with dihydropyridines. No differences at rest, but striking increases of the low-frequency component of systolic arterial pressure variability were observed in nicardipine-treated patients during both standardized excitatory stimuli, suggesting a marked increase in sympathetic vasomotor drive. As to reference treatments, patients treated with atenolol displayed the lowest values, and patients treated with cilazapril (for 4 weeks) provided intermediate values. In conclusion, shorter acting dihydropyridine calcium channel antagonists may induce an exaggerated increase in sympathetic vasomotor drive during standardized laboratory stressors. PMID- 9213213 TI - Differential inhibition of plasma versus tissue ACE by utibapril: biochemical and functional evidence for inhibition of vascular ACE activity. AB - Utibapril is an angiotensin-converting enzyme (ACE) inhibitor with a proposed tissue-specific inhibitory profile. This implies that at a certain dose, utibapril should be able to inhibit tissue ACE activity without affecting plasma ACE. Moreover, if tissue ACE activity is rate limiting, functional conversion of angiotensin I should be decreased. Accordingly, we studied the dose-dependent effect of long-term treatment with utibapril on plasma and tissue ACE. Normal Wistar rats were randomly allocated to oral treatment with different doses of utibapril (0, 2, 10, 50, or 250 micrograms/kg/day) for 30 days. Tissue inhibition of ACE was assessed biochemically, whereas functional conversion of angiotensin I was determined in the isolated organ. Utibapril significantly inhibited plasma, renal, and vascular ACE but not ventricular ACE activity. Notably, however, only treatment with the highest dose of utibapril resulted in a significant inhibition of plasma ACE, whereas vascular ACE activity was already significantly inhibited after treatment with a lower dose of utibapril. In accordance, utibapril dose dependently inhibited the contraction of isolated aortic rings to angiotensin I. Furthermore, angiotensin I-induced decreases in coronary flow in the isolated heart were significantly inhibited after treatment with the higher doses of utibapril. These data suggest the preferential inhibition of vascular ACE by utibapril in normal rats. Furthermore, the dose-dependent inhibition of the functional conversion of angiotensin I indicates that the tissue ACE activity may be rate limiting in vascular beds in rats. PMID- 9213214 TI - Effects of the novel T-type calcium channel antagonist mibefradil on human myocardial contractility in comparison with nifedipine and verapamil. AB - Mibefradil (Ro 40-5967) is a novel nondihydropyridine calcium antagonist. The aim of our study was to compare the negative inotropic effects of the well-known 1,4 dihydropyridine nifedipine and the phenylalkylamine verapamil with those of mibefradil. Isometric force of contraction in response to these substances was determined in isolated, electrically driven left ventricular papillary muscle strips from failing human hearts (1 Hz, 37 degrees C). The hearts were obtained during cardiac transplantation (n = 9) and mitral valve-replacement operations (n = 9). The calcium antagonists studied significantly (p < 0.05) depressed basal force of contraction in a concentration-dependent manner. The effect started at concentrations > 0.001 microM for nifedipine and > 0.01 microM for verapamil, but only at concentrations > 10 microM for mibefradil. Only in the presence of nifedipine and verapamil was a significant rightward shift of the inotropic concentration--response curves to calcium and a depression of the maximal effects of calcium observed. With respect of the relation between the therapeutic active plasma concentration in vivo and the negative intropic potency in vitro, it became evident that the difference between therapeutically beneficial concentrations and potentially hazardous cardiodepressant activity increases from nifedipine to mibefradil. We conclude that this new generation of calcium antagonists, almost lacking cardiodepressant effects, could lead to a greater therapeutic index and greater safety in the treatment of cardiovascular diseases. PMID- 9213215 TI - Role of the glycolytic protein, glyceraldehyde-3-phosphate dehydrogenase, in normal cell function and in cell pathology. AB - The glycolytic protein glyceraldehyde-3-phosphate dehydrogenase (GAPDH) appeared to be an archtypical protein of limited excitement. However, independent studies from a number of different laboratories reported a variety of diverse biological properties of the GAPDH protein. As a membrane protein, GAPDH functions in endocytosis; in the cytoplasm, it is involved in the translational control of gene expression; in the nucleus, it functions in nuclear tRNA export, in DNA replication, and in DNA repair. The intracellular localization of GAPDH may be dependent on the proliferative state of the cell. Recent studies identified a role for GAPDH in neuronal apoptosis. GAPDH gene expression was specifically increased during programmed neuronal cell death. Transfection of neuronal cells with antisense GAPDH sequences inhibited apoptosis. Lastly, GAPDH may be directly involved in the cellular phenotype of human neurodegenerative disorders, especially those characterized at the molecular level by the expansion of CAG repeats. In this review, the current status of ongoing GAPDH studies are described (with the exception of its unique oxidative modification by nitric oxide). Consideration of future directions are suggested. PMID- 9213217 TI - Heat shock protein 27 stimulates recovery of RNA and protein synthesis following a heat shock. AB - Constitutive expression of human hsp27 resulted in a 100-fold increase in survival to a single lethal heat shock in CHO cells without effecting the development of thermotolerance. A possible mechanism for the thermoprotective function of hsp27 may be increased recovery of protein synthesis and RNA synthesis following a heat shock. A lethal heat shock (44 degrees C, 30 min) results in a 90% reduction in the rate of protein synthesis in non-tolerant cells. Control transfected cells recovered protein synthesis to a pre-heat shock rate 10 h after the heat shock; while cell lines that constitutively express human hsp27 recovered 6 h after the heat shock. Thermotolerant cells had a 50% reduction in protein synthesis, which recovered within 7 h following the heat shock. The same lethal heat shock (44 degrees C, 30 min) reduced RNA synthesis by 60% in the transfected cell lines, with the controls recovering in 7 h; while the hsp27 expressing cell lines recovered within 5 h. Thermotolerant cells had a 40% reduction in RNA synthesis and were able to recover within 4 h. The enhanced ability of hsp27 to facilitate recovery of protein synthesis and RNA synthesis following a heat shock may provide the cell with a survival advantage. PMID- 9213216 TI - Post-proliferative cyclin E-associated kinase activity in differentiated osteoblasts: inhibition by proliferating osteoblasts and osteosarcoma cells. AB - Spontaneous differentiation of normal diploid osteoblasts in culture is accompanied by increased cyclin E associated kinase activity on (1) the retinoblastoma susceptibility protein pRB, (2) the p107 RB related protein, and (3) two endogenous cyclin E-associated substrates of 78 and 105 kD. Activity of the differentiation-related cyclin E complexes (diff.ECx) is not recovered in cdc2 or cdk2 immunoprecipitates. Phosphorylation of both the 105 kD endogenous substrate and the p107 exogenous substrate is sensitive to inhibitory activity (diff.ECx-i) present in proliferating osteoblasts. This inhibitory activity is readily recruited by the cyclin E complexes of differentiated osteoblasts but is not found in cyclin E immunoprecipitates of the proliferating cells themselves. Strong inhibitory activity on diff.ECx kinase activity is excerted by proliferating ROS 17/2.8 osteosarcoma cells. However, unlike the normal diploid cells, the diff.ECx-i activity of proliferating ROS 17/2.8 cells is recovered by cyclin E immunoprecipitation. The cyclin-dependent kinase inhibitor p21CIP1/WAF1 inhibits diff.ECx kinase activity. Thus, our results suggest the existence of a unique regulatory system, possibly involving p21CIP1/WAF1, in which inhibitory activity residing in proliferating cells is preferentially targeted towards differentiation-related cyclin E-associated kinase activity. PMID- 9213218 TI - Polyamine-dependent alterations in the structure of microfilaments, Golgi apparatus, endoplasmic reticulum, and proteoglycan synthesis in BHK cells. AB - The activity of ornithine decarboxylase, the key enzyme in the synthesis of polyamines, is essential for proliferation and differentiation of all living cells. Two inhibitors of ornithine decarboxylase, alpha-difluoromethylornithine (DFMO) and 1-aminooxy-3-aminopropane (APA), caused swelling of endoplasmic reticulum (ER) and medial and trans Golgi cisternae, and the disappearance of stress fibers, as visualized by staining with fluorescent concanavalin A (ConA), C6-NBD-ceramide or wheat germ agglutinin (WGA), and phalloidin, respectively. In contrast, the pattern of microtubules, stained with a beta-tubulin antibody, was not affected. Rough ER seemed to be especially affected in polyamine deprivation forming whorls and involutions, which were observed by transmission electron microscopy. Since ER and Golgi apparatus are vital parts of the glycosylation and secretory machinery of the cell, we tested the ability of these structurally altered cell organelles to synthesize proteoglycans using [3H]glucosamine and [35S]sulfate as precursors. The total incorporation rate into proteoglycans and hyaluronan was not reduced in polyamine-deprived cells, suggesting that the total glycosylation capacity of cells was not affected. However, the synthesis of a high molecular weight proteoglycan containing chondroitin and keratan sulfate was completely inhibited. The remodeling of cytoskeleton and rough endoplasmic reticulum in polyamine deprivation may perturb the synthesis and secretion of the components of membrane skeleton and of the extracellular matrix, e.g., proteoglycans. Rough ER and cytoskeleton may be the targets where polyamines affect cell proliferation and differentiation. PMID- 9213220 TI - Expression of the vitamin D and the retinoid X receptors in Saccharomyces cerevisiae: alternative in vivo models for ligand-induced transactivation. AB - The transcription factors of the nuclear hormone receptor family regulate gene expression via a complex network of macromolecular interactions. The ligand dependent activity of the vitamin D receptor is of particular interest because it modulates gene expression by the heterodimeric interaction with retinoid X receptors. We report here that individual functions of the vitamin D receptor including DNA-binding, homo- and heterodimerization and transactivation can be reconstituted in the yeast Saccharomyces cerevisiae. Interestingly, the simultaneous expression of the native vitamin D receptor and the retinoid X receptor beta resulted in a ligand independent transactivation of the lacZ reporter gene coupled to a mouse osteopontin vitamin D response element. However, homodimerization of the vitamin D receptor and heterodimerization were strongly enhanced upon ligand binding, when the receptors were expressed as fusion proteins with the Gal4 transcription factor in a yeast two-hybrid system. Furthermore, transactivating activity of a Gal4-fused vitamin D receptor was induced by vitamin D in a one-hybrid system devoid of retinoid X receptors. In addition, both Gal4-based systems behaved similar with regard to their dose dependent response to vitamin D and related compounds when compared to the transcriptional activity of the vitamin D receptor in transiently transfected MCF 7 cells. Our results point out that specific ligands strongly enhanced receptor dimerization and induced transactivation in yeast and in MCF-7 cells. The constitutive transactivation by vitamin D receptor-retinoid X receptor heterodimers in yeast, depending on DNA binding of the receptors, strongly argues for the existence of cofactors, which are absent in yeast, but play a fundamental role in gene regulation in higher eukaryotic organisms. PMID- 9213219 TI - Phosphorylation of the oncogenic transcription factor interferon regulatory factor 2 (IRF2) in vitro and in vivo. AB - IRF2 is a transcription factor, possessing oncogenic potential, responsible for both the repression of growth-inhibiting genes (interferon) and the activation of cell cycle-regulated genes (histone H4). Surprisingly little is known about the post-translational modification of this factor. In this study, we analyze the phosphorylation of IRF2 both in vivo and in vitro. Immunoprecipitation of HA tagged IRF2 expressed in 32P-phosphate labelled COS-7 cells demonstrates that IRF2 is phosphorylated in vivo. Amino acid sequence analysis reveals that several potential phosphorylation sites exist for a variety of serine/threonine protein kinases, including those of the mitogen activated protein (MAP) kinase family. Using a battery of these protein kinases we show that recombinant IRF2 is a substrate for protein kinase A (PKA), protein kinase C (PKC), and casein kinase II (CK2) in vitro. However, other serine/threonine protein kinases, including the MAP kinases JNK1, p38, and ERK2, do not phosphorylate IRF2. Two-dimensional phosphopeptide mapping of the sites phosphorylated by PKA, PKC, and CKII in vitro demonstrates that these enzymes are capable of phosphorylating IRF2 at multiple distinct sites. Phosphoaminoacid analysis of HA-tagged IRF2 immunoprecipitated from an asynchronous population of proliferating, metabolically phosphate labelled cells indicates that this protein is phosphorylated exclusively upon serine residues in vivo. These results suggest that the oncogenic protein IRF2 may be regulated via multiple pathways during cellular growth. PMID- 9213221 TI - Identification of mu-, m-calpains and calpastatin and capture of mu-calpain activation in endothelial cells. AB - The presence of the calpain-calpastatin system in human umbilical vein endothelial cells (HUVEC) was investigated by means of ion exchange chromatography, Western blot analysis, and Northern blot analysis. On DEAE anion exchange chromatography, calpain and calpastatin activities were eluted at approximately 0.30 M and 0.15-0.25 M NaCl, respectively. For half-maximal activity, the protease required 800 microM Ca2+, comparable to the Ca2+ requirement of m-calpain. By Western blot analysis, the large subunit of mu calpain (80 kDa) was found to be eluted with calpastatin (110 kDa). Both the large subunit of m-calpain (80 kDa) and calpastatin were detected in the respective active fractions. By Northern blot analysis, mRNAs for large subunits of mu- and m-calpains were detected in single bands, each corresponding to approximately 3.5 Kb. Calpastatin mRNA was observed in two bands corresponding to approximately 3.8 and 2.6 Kb. Furthermore, the activation of mu-calpain in HUVEC by a calcium ionophore was examined, using an antibody specifically recognizing an autolytic intermediate form of mu-calpain large subunit (78 kDa). Both talin and filamin of HUVEC were proteolyzed in a calcium-dependent manner, and the reactions were inhibited by calpeptin, a cell-permeable calpain specific inhibitor. Proteolysis of the cytoskeleton was preceded by the appearance of the autolytic intermediate form of mu-calpain, while the fully autolyzed postautolysis form of mu-calpain (76 kDa) remained below detectable levels at all time points examined. These results indicate that the calpain-calpastatin system is present in human endothelial cells and that mu-calpain may be involved in endothelial cell function mediated by Ca2+ via the limited proteolysis of various proteins. PMID- 9213222 TI - Localization of silencer and enhancer elements in the human type X collagen gene. AB - Collagen type X is a short, network-forming collagen expressed temporally and spatially tightly controlled in hypertrophic chondrocytes during endochondral ossification. Studies on chicken chondrocytes indicate that the regulation of type X collagen gene expression is regulated at the transcriptional level. In this study, we have analyzed the regulatory elements of the human type X collagen (Col10a1) by reporter gene constructs and transient transfections in chondrogenic and nonchondrogenic cells. Four different promoter fragments covering up to 2,864 bp of 5'-flanking sequences, either including or lacking the first intron, were linked to luciferase reporter gene and transfected into 3T3 fibroblasts, HT1080 fibrosarcoma cells, prehypertrophic chondrocytes from the resting zone, hypertrophic chondrocytes, and chondrogenic cell lines. The results indicated the presence of three regulatory elements in the human Col10a1 gene besides the proximal promoter. First, a negative regulatory element located between 2.4 and 2.8 kb upstream of the transcription initiation site was active in all nonchondrogenic cells and in prehypertrophic chondrocytes. Second, a positive, but also non-tissue-specific positive regulatory element was present in the first intron. Third, a cell-type-specific enhancer element active only in hypertrophic chondrocytes was located between -2.4 and -0.9 kb confirming a previous report by Thomas et al. [(1995): Gene 160:291-296]. The enhancing effect, however, was observed only when calcium phosphate was either used for transfection or included in the culture medium after lipofection. These findings demonstrate that the rigid control of human Col10a1 gene expression is achieved by both positive and negative regulatory elements in the gene and provide the basis for the identification of factors binding to those elements. PMID- 9213223 TI - Levels of DNA strand breaks and superoxide in phorbol ester-treated human granulocytes. AB - Phorbol ester treatment of granulocytes triggers release of superoxide (O2.-) and a concomitant burst of DNA strand breaks. The relationship between the amount of O2.- and the number of DNA breaks has not previously been explored. To quantify the relatively large amount of O2.- generated over a 40-min period by 1 x 10(6) granulocytes/mL, a discontinuous "10-min pulse" method employing cytochrome c was used; 140 nmol O2.- per 1 x 10(6) cells was detected. DNA strand breaks were quantified by fluorimetric analysis of DNA unwinding (FADU). To vary the level of O2.- released by cells, inhibitors of the respiratory burst were used. Sodium fluoride (1-10 mM) and staurosporine (2-10 nM) both inhibited O2.- production. In both cases, however, inhibition of strand breakage was considerably more pronounced than inhibition of O2.-. Zinc chloride (50-200 microM) inhibited both O2.- and DNA breaks, approximately equally. Dinophysistoxin-1 (okadaic acid) inhibited O2.- production more effectively than it inhibited DNA breaks. O2.- dismutes to H2O2, a reactive oxygen species known to cause DNA breaks. The addition of catalase to remove extracellular H2O2 had no effect on DNA breakage. Using pulse field gel electrophoresis, few double-stranded breaks were detected compared to the number detected by FADU, indicating that about 95% of breaks were single-stranded. The level of DNA breaks is not directly related to the amount of extracellular O2.- or H2O2 in PMA-stimulated granulocytes. We conclude that either an intracellular pool of these reactive oxygen species is involved in breakage or that the metabolic inhibitors are affecting a novel strand break pathway. PMID- 9213224 TI - Expression of human p140trk receptors in p140trk-deficient, PC12/endothelial cells results in nerve growth factor-induced signal transduction and DNA synthesis. AB - Nerve growth factor (NGF) regulates proliferation, differentiation, and survival of sympathetic and sensory neurons through the tyrosine kinase activity of its receptor, p140trk. These biological effects of NGF depend upon the signal mediating function of p140trk substrates which are likely to differ from cell to cell. To define p140trk receptor substrates and the details of signalling by NGF in the hybrid cell PC12EN, we stably transfected cultures with a vector encoding a full-length human p140trk cDNA sequence. Two stably transfected clones, one expressing p140trk with higher affinity (PC12EN-trk3; Kd 57.4 pM, Bmax 9.7 pmole/mg) and one expressing p140trk with a lower affinity (PC12EN-trk1; Kd 392.4 pM, Bmax 5.7 pmole/mg) were generated. Radioreceptor assays indicate that transfected p140trk receptors show slow NGF-dissociation kinetics, are resistant to trypsin or Triton X-100 treatment, are specific for NGF compared to other neurotrophins, and are internalized or downregulated as are native PC12 p140trk receptors. NGF stimulates p140trk tyrosine phosphorylation in a dose- (0.01-10 ng/ml) and time- (5-120 min) dependent manner, and tyrosine phosphorylation was inhibited by 200-1,000 nM K-252a. NGF-induced Erk stimulation for 60 min was assessed using myelin basic protein as a substrate. NGF treatment also led to an increased phosphorylation of p70S6k, SNT, and phospholipase C gamma, demonstrating that the major NGF-stimulated signalling pathways found in other cells are activated in PC12EN-trk cells. Staurosporine (5-50 nM) rapidly and dBcAMP (1 mM) more slowly, but not NGF induced morphological differentiation in PC12EN-trk cells. Rather, NGF treatment in low-serum medium stimulated a 1.3- and 2.3-fold increase in DNA synthesis measured by [3H]thymidine incorporation in PC12EN-trk1 and PC12EN-trk3, respectively. These data highlight the functionality of the transfected p140trk receptors and indicate that these transfected cells may serve as a novel cellular model facilitating the study of the mitogenic properties of NGF signalling and the transducing role of the p140trk receptor substrates. PMID- 9213225 TI - Human papillomavirus (HPV) 16 E6 sensitizes cells to atractyloside-induced apoptosis: role of p53, ICE-like proteases and the mitochondrial permeability transition. AB - Infection of cervical epithelial cells with certain high risk HPV genotypes is thought to play an etiologic role in the development of cervical cancer. In particular, HPV type 16 and 18 early protein 6 (E6) is thought to contribute to epithelial transformation by binding to the tumor suppressor protein p53, targeting it for rapid proteolysis, resulting in loss of its cell cycle arrest and apoptosis-inducing activities. Recent data indicate that factors responsible for triggering apoptosis reside in the cytoplasm of cells, and not in the nucleus. In particular, the findings that mitochondria are required in certain cell-free models for induction of apoptosis and that bcl-2 is localized to mitochondria have focused attention on the role of the mitochondrial membrane permeability transition (MPT) in apoptosis. Here we present data to indicate that HPV 16 E6 expression sensitizes cells to MPT-induced apoptosis. We also report that HPV 16 E6 sensitization of cells to MPT-induced apoptosis occurs only in the presence of wildtype (wt) p53 expression. The extent of apoptosis induced by atractyloside (an inducer of the MPT) in normal, temperature-sensitive (ts) p53, and HPV-16 E6 transfected J2-3T3 cells, and the HPV expressing cervical carcinoma cell lines SiHa, Hela and CaSki was determined. C33A cells, which express mutant p53 but not HPV, were also exposed to atractyloside in the presence or absence of HPV 16 E6 expression. Dose-dependent apoptosis induced by atractyloside in normal J2-3T3 cells and cervical carcinoma cells was measured by loss of cell viability, nuclear fragmentation and DNA laddering. The sensitivity of cells to atractyloside-induced apoptosis was found to be: HPV 16 E6-J2-3T3 > CaSki > normal-J2-3T3 cells approximately ts p53-J2-3T3 approximately vector-J2-3T3 cells > Hela > SiHa > C33A approximately C33A 16 E6. Cyclosporin A (CsA), an inhibitor of the MPT, and ICE-I, a protease inhibitor, provided protection against atractyloside-induced apoptosis. These findings indicate that: 1) high risk HPV 16 E6 protein is capable of sensitizing cells to apoptosis; 2) HPV 16 E6 sensitization of cells to atractyloside-induced apoptosis occurs in a p53 dependent fashion; 3) the target of HPV 16 E6 sensitization of cells to atractyloside-induced apoptosis is the mitochondria; and 4) HPV 16 E6 sensitization of cells to atroctycoside-induced apoptosis involves an ICE-like protease-sensitive mechanism, regulating the onset of the MPT. These findings constitute the first evidence that mitochondria play a role in HPV 16 E6 modulation of apoptosis. PMID- 9213226 TI - Use of lanthanum to accurately quantify insulin-like growth factor binding to proteins on cell surfaces. AB - Insulin-like growth factor binding proteins (IGFBPs) are found both associated with cells and in extracellular fluids. Cell-associated IGFBPs increase [125I] IGF binding to cell monolayers, whereas extracellular (soluble, released) IGFBPs decrease binding. In the current study, we show that either IGFBP-3 or IGFBP-5 are the major forms of IGFBP released from monolayers of human GM10 fibroblasts, T98G glioblastoma cells and forskolin-treated bovine MDBK cells. IGFBPs represent the most abundant [125I]-IGF-I binding site on GM10 and T98G cell monolayers, but 4-17% of the total cell-associated IGFBPs are released from the cell monolayer at 8 degrees C during their quantification. Most of the IGFBPs (> 70%) are released from MDBK cells. Quantitative estimates of [125I]-IGF binding to the cell monolayers are altered because of the ability of the released IGFBPs to reduce the amount of radiolabeled ligand that is available to bind to the cell surface. Lanthanum (La3+) depresses IGFBP release from all three cell types (> 80% for GM10 and T98G cells and > 65% for MDBK cells). The effect was cation specific, noted with La3+ or Zn2+ but not with either Mn2+, Sr2+ or Se3+. The effect was also IGFBP specific; La3+ markedly depressed the release of IGFBP-3 and IGFBP-5, but had less of an effect on IGFBP-2 and IGFBP-4. Concomitant with a decrease in IGFBP-3 and IGFBP-5 release, La3+ caused an increase in [125I]-IGF-I binding to cell-associated IGFBPs and type I IGF receptors. The released soluble IGFBPs have a three- to 20-fold greater affinity (Ka) for [125I]-IGF-I compared to cell associated IGFBPs. La3+ did not alter the affinity constants of cell-associated IGFBPs. In summary, we have identified a means to prevent loss of IGFBPs from cell monolayers during binding assays. This procedure will be useful in accurately quantifying the levels of IGFBPs on cell monolayers and in determining the role of cell-associated IGFBPs in controlling IGF activity. Retention of cell associated low affinity IGFBPs may be important in controlling the size of the pericellular IGF pool and in regulating IGF-I access to the type IIGF receptor. PMID- 9213227 TI - Phosphorylation of the G protein alpha-subunit, G alpha 2, of Dictyostelium discoideum requires a functional and activated G alpha 2. AB - The G alpha 2-subunit of Dictyostelium discoideum is essential to the initial stage of the cell's developmental life cycle. In response to the extracellular chemoattractant, cAMP, G alpha 2 is activated and transiently phosphorylated on serine-113 [Chen et al. (1994): J Biol Chem 269:20925-20930]. The role of G alpha 2 phosphorylation remains elusive; cells expressing the S113A, nonphosphorylated mutation of G alpha 2 appear to proceed through the developmental phase normally. To gain insight into the function of G alpha 2 phosphorylation, the conditions for G alpha 2 phosphorylation were examined using a variety of alpha-subunit point mutations and chimeras. Mutations that block the G protein activation cycle prior to or at the hydrolysis of GTP (G alpha 2-S45A, G alpha 2-G207A, and G alpha 2-Q208L) preclude G alpha 2 phosphorylation in vivo. Phosphorylation of the G alpha 2-Q208L mutation does however occur in an in vitro phosphorylation assay. It appears that G alpha 2 phosphorylation, shown previously in vivo to require the cAMP receptor, also requires signaling through the G2 pathway. Results from the in vitro assay suggest that the substrate for phosphorylation is the alpha subunit monomer. PMID- 9213228 TI - Biophysical bases for delayed and aberrant motor development in young children with achondroplasia. AB - Achondroplasia, the most common skeletal dysplasia, is characterized by delayed and aberrant motoric development in childhood. Delays and aberrancy are secondary to the anatomical differences inherent in people with achondroplasia. We present a photographic essay documenting biophysical differences, aberrant pre-orthograde movement strategies, and selected adaptive techniques. A parental questionnaire assessed the presence of, predominance, and ages at which various types of pre orthograde locomotion were observed. Fine and gross motor skills were assessed contemporaneously by use of the Denver Developmental Screening Test in 93 children with achondroplasia and were found to be more delayed than previously reported. Physicians, therapists, early-childhood educators, and parents should recognize that aberrant does not mean maladaptive and that different development is not defective development. PMID- 9213229 TI - Primitive reflexes and early motor development. AB - To investigate the relationship between primitive reflexes and typical early motor development, 156 full-term infants with normal 18-month developmental outcomes were assessed using a modified Primitive Reflex Profile (PRP) and the Alberta Infant Motor Scale (AIMS) at 6 weeks and 3 and 5 months. No significant positive or negative correlations were obtained between the scores of the PRP and the AIMS at any of the ages assessed. Similarly, PRP scores did not differ between infants scoring above and below the 50th percentile on the AIMS. Primitive reflexes were unrelated to motor development. If this finding is maintained among infants at risk for motor disability, observational assessment of spontaneously generated movement, rather than isolated testing of primitive reflexes, might yield more valuable information on the child's overall level of maturation. Intervention for children with identified motor delays or neurological impairments might not need to be focused on either suppression or enhancement of these motor functions. PMID- 9213230 TI - Adolescent girls' perceived prevalence of sexually transmitted diseases and condom use. AB - Little is known concerning sexually experienced and inexperienced adolescent girls' perceptions of the prevalence of condom use and sexually transmitted diseases (STDs). Girls (n = 174; 41% sexually experienced) rated the prevalence of condom use among friends and STDs among male and female friends and adolescents in general. Girls perceive the prevalence of STDs similarly across both gender and level of familiarity. For the most part, however, the girls perceived the prevalence among boys and girls more similarly than among friends and adolescents in general. No significant differences were found between sexually experienced and inexperienced girls in perceptions of condom use prevalence, but girls with a history of STD perceived condoms as used less frequently. Girls with an STD history perceived STDs as the most prevalent, followed by sexually inexperienced girls and then sexually experienced girls without a history of an STD. After an adolescent girl initiates sexual intercourse, STD experience could be a key variable in affecting her perceptions. Prevention programs can incorporate an understanding of patients' perceptions of condom use and STDs. PMID- 9213231 TI - Infant formula quiets crying human newborns. AB - Milk (Similac), sucrose (12% wt/vol), or water were delivered to crying normal newborns once per minute for 5 minutes, in a volume of 0.1 mL/delivery. Milk and sucrose markedly reduced infant crying, and this calm persisted during the 3 minutes after substance delivery. Infants who received water were only marginally quieted, and this calm did not persist. Despite quieting agitated infants, milk did not cause them to bring their hands to their mouths during the period of milk treatment, whereas infants who received sucrose did bring their hands to their mouths. These data demonstrate that milk effectively quiets human newborns, that its quieting effects endure, and that the mechanisms that quiet and that underlie hand-in-mouth engagement are separable and independent. PMID- 9213232 TI - Environmental factors influencing weaning of a young child from mechanical ventilator support. AB - We examined the contribution of physiologic and environmental variables to the process of weaning a child with chronic respiratory failure from mechanical ventilation support. Surveillance measures, e.g., blood oxygenation, were obtained from a 6-year-old child with multiple medical and developmental disorders who received three different rates (24, 22, and 20 tidal volumes per minute) of intermittent mechanical ventilation. Direct observations were used to calculate rates of aberrant behavior, e.g., aggression toward self, for task versus play settings within the intermittent mechanical ventilation rates. Rates of aberrant behavior and adult responses were tabulated from videotaped observations for task, attention, and no attention settings. The greatest rate of aberrant behavior occurred during tasks compared with play activities, regardless of whether attention was provided while playing. Adults also responded more often to aberrant behaviors during task versus play conditions. Clinical implications are discussed concerning the inclusion of developmental and behavioral variables during weaning from mechanical ventilation. PMID- 9213233 TI - Diagnostic and Statistical Manual for Primary Care (DSM-PC) Child and Adolescent Version: design, intent, and hopes for the future. PMID- 9213234 TI - DSM-PC: bridging pediatric primary care and mental health services. PMID- 9213235 TI - DSM-PC: experiences in primary care practice and resident education. PMID- 9213236 TI - Use of the DSM-PC and implications for reimbursement. PMID- 9213237 TI - Sammy: gender identity concerns in a 6-year-old boy. PMID- 9213238 TI - A review of behavioral screening practices in pediatric settings: do they pass the test? AB - We reviewed the current status of behavior screening methods, such as parental questionnaires, for identifying behavioral problems in children seen in pediatric settings. Information is organized around basic criteria for implementing screening procedures. We conclude that although use of parent-completed questionnaires, such as the Child Behavior Checklist and the Pediatric Symptom Checklist, can increase identification of child behavioral dysfunction in pediatric settings, it is unclear whether screening will cause a change in physician behaviors necessary to improve child functional outcomes. Clinical and research implications are discussed. PMID- 9213239 TI - Significance of Helicobacter pylori infection as a risk factor in gastric cancer: serological and histological studies. AB - We conducted a case-control study to examine the association of Helicobacter pylori infection as a risk factor in gastric cancer in the Japanese population. Serum IgG antibodies for Helicobacter pylori were determined in 55 consecutive patients with gastric cancer and in 75 age- and sex-matched mass survey subjects and 57 age- and sex-matched cancer-free patients with conditions considered at a high risk for development of gastric cancer (precancerous condition). We examined the histology in all subjects and particular focus was placed on the extent of Helicobacter pylori-associated gastritis. The seroprevalence of Helicobacter pylori in gastric cancer patients (82%) and those with a precancerous condition (89%) was significantly higher (P < 0.005) than that in the mass survey subjects (60%). Positive relative risk associations were found for patients with gastric cancer (odds ratio, 3, with 95% confidence intervals of 1.69-5.33) and those with a precancerous condition (odds ratio, 5.66, with 95% confidence intervals 2.66 12.03). Significant differences were found when comparisons were made among the case-control groups who were H. pylori-positive and had inflammatory cell infiltration (P = 0.0127). The characteristics of Helicobacter pylori in histologically examined gastric mucosa showed differences between Helicobacter pylori-infected and uninfected persons in all groups. However, for none of these groups was there a significant differences between background mucosa for Helicobacter pylori-infected persons with or without gastric cancer. Helicobacter pylori seroprevalence is strongly associated with an increased risk of gastric cancer and with a precancerous condition; histological investigation did not define additional factors that might be associated with increased cancer risk. PMID- 9213240 TI - Strain differences in the induction of intestinal metaplasia by X-irradiation in rats. AB - Strain differences in the susceptibility of rats to induction of intestinal metaplasia by X-irradiation were examined. The gastric regions of 5-week-old males of five inbred strains of rats (F344/NSlc, Copenhagen, Buffalo/NacJcl, and ACI/NHos) and three strains of randomly bred rats (HOS:Donryu, Slc:Wistar, Slc:SD) were irradiated with a total dose of 20 Gy X-ray given in two equal fractions at 3-day interrals. When examined after the rats were killed, 6 months after the last irradiation, the number of intestinal metaplastic crypts positive for alkaline phosphatase (ALP) was highest in the Donryu and lowest in the Copenhagen rats. Morphologically, the number of crypts with intestinal metaplasia in the glandular stomachs of Donryu, Wistar, SD, and Buffalo rats was higher than the number in ACI, F344, and Copenhagen rats. Intestinal metaplasia was more frequently observed in the pyloric than in the fundic glands. These results demonstrate that the induction of intestinal metaplasia by X-irradiation in rats is greatly influenced by the rat strain. PMID- 9213242 TI - Analysis of bile acids in colon residual liquid or fecal material in patients with colorectal neoplasia and control subjects. AB - Bile acids are believed to play a role in the etiology of colorectal cancer. To examine the relationship between bile acids and colorectal neoplasia, bile acids in colon residual liquid or fecal material were analyzed in 18 patients with colorectal adenoma, 12 patients with colorectal cancer, and 18 healthy control subjects. High-performance liquid chromatography combined with immobilized 3 alpha-hydroxysteroid dehydrogenase in column form showed a significant elevation in the proportion of deoxycholic acid (P < 0.05), lithocholic acid (P < 0.05), secondary bile acids (deoxycholic acid plus lithocholic acid) (P < 0.02), and the chenodeoxycholic acid-lithocholic acid family (chenodeoxycholic acid plus lithocholic acid) (P < 0.05) in the colon residual liquid or fecal material of the patients with colorectal adenoma compared with proportions in the control subjects. A similar trend was noted in the patients with colorectal cancer compared to the control subjects. These findings suggested that an increase in the proportion of secondary bile acids, in particular, of lithocholic acid, was closely related to the pathogenesis of colorectal neoplasia. PMID- 9213243 TI - Estimation of serum beta-2-microglobulin in children with malabsorption disorders syndrome. AB - The aim of this study was to examine serum levels of beta-2-microglobulin (b-2-m) in 132 children at various stages for the evaluation of celiac disease (CD). Serum b-2-m was analyzed by a radio immunoassay (RIA) method, using a beta-2 micro RIA kit (Pharmacia, Uppsala, Sweden). The mean concentration of b-2-m in children with an established diagnosis of CD was 4.38 +/- 1.86 mg/l. In children receiving a gluten-free diet, the mean b-2-m concentration was 1.95 +/- 1.09 mg/l, and in children who received a gluten-containing diet, the concentration was 3.19 +/- 0.71 mg/l. In children with CD who were on a gluten-free diet and who presented no antibodies against EmA in class IgA serum, b-2-m concentration was within the normal range (1.86 +/- 0.55 mg/l). The concentration of b-2-m in children with secondary malabsorption syndrome was within the physiological range (1.77 +/- 0.64 mg/l). In children with IgA-EmA antibodies present in serum, the b 2-m concentration was significantly higher (3.5 +/- 1.23 mg/l; P < 0.001) than that in children with IgA-EmA in serum. We showed a linear dependence between the degree of villous atrophy in CD and concentrations of b-2-m in serum (r2 = 0.94). Determination of b-2-m concentration in sera of children with CD may be used to monitor treatment with a gluten-free diet and to differentiate secondary malabsorption syndrome from CD. PMID- 9213241 TI - Trophic effects of glicentin on rat small-intestinal mucosa in vivo and in vitro. AB - To define the role of glicentin the active site of enteroglucagon, we evaluated the trophic effects of recombinant rat glicentin on rat small intestine and IEC-6 cells. In vivo, a significant increase was observed in jejunal wet weight, protein content, DNA content, and alkaline phosphatase activity after the subcutaneous administration of 100 micrograms/kg per day of glicentin for 2 weeks. In the ileum, however, there were no significant differences between the control versus glicentin groups in any of these parameters. Ornithine decarboxylase (ODC) activity 3.5 h after an intraperitoneal injection of glicentin was increased in the jejunal mucosa, but not in the ileal mucosa. In vitro, glicentin, at a dose of more than 100 ng/ml, significantly increased both tritium-thymidine incorporation and the number of IEC-6 cells. These findings indicate that glicentin exerts direct trophic effects on the rat small-intestinal mucosa and on the rat small-intestinal cell line, IEC-6, and that this peptide appears to be an active site of enteroglucagon. PMID- 9213244 TI - Red cell survival in patients with nonalcoholic liver cirrhosis before and after distal splenorenal shunt. AB - We previously reported severe hemolysis in one patient immediately after distal splenorenal shunt (DSRS). The purpose of the present study was to evaluate changes in red cell survival after DSRS. In ten patients with nonalcoholic cirrhosis in whom DSRS was performed for esophageal varices, red cell survival and splenic quantitative hemodynamic studies were performed before and after DSRS. The splenic venous blood flow per unit volume (flow/volume ratio) was calculated. The red cell survival was significantly (P < 0.05) shortened after DSRS; the apparent half-life survival time (T 1/2) before and after DSRS was 24.6 +/- 5.9 (mean +/- SD) and 16.3 +/- 8.5 days, respectively. After DSRS, the spleen volume was significantly (P < 0.05) decreased, whereas the splenic venous blood flow was slightly increased. The spleen flow/volume ratio was significantly (P < 0.05) increased after DSRS. There was a significant and negative correlation (r = -0.684, P < 0.05) between the postoperative percentage change in T 1/2 and the spleen flow/volume ratio. These findings suggest that the red cell survival period is significantly decreased, after DSRS in patients with nonalcoholic cirrhosis, and that the increased splenic blood flow per unit spleen volume after DSRS may play an important role in the hemolytic reaction in the spleen after this procedure. PMID- 9213245 TI - Immunological evaluation in oral lichen planus with chronic hepatitis C. AB - Oral lichen planus (OLP) is frequently associated with hepatitis C virus (HCV) infection. We have previously reported that the pathogenesis of OLP arises from host rather than viral factors. In this study, we investigated the role of these factors in 41 patients with chronic hepatitis C: 22 with OLP (group 1) and 19 without OLP (group 2). All patients had antibodies to HCV (anti-HCV) and were serum HCV RNA-positive; none were HBsAg-positive. Immune abnormalities in serum were evaluated by testing for antinuclear antibody (ANA), rheumatoid factor (RF) activity, immunoglobulins (IgG, IgM, and IgA), and cryoglobulin. The rate for ANA positivity and IgM levels were significantly higher in group 1 than in group 2 (P < 0.05). Mean age in group 1 was significantly higher in group 2 (P = 0.0001). Of the factors tested, ANA, IgM, and age, logistic regression showed that age correlated independently with OLP (P = 0.003). In 5 patients in group 1, the infiltrating lymphocytic subsets of the OLP lesion were examined histopathologically. Predominant T cell infiltration was shown in all 5 patients. In addition to host factors, wer also examined viral factors in both groups of patients, measuring serum HCV RNA level and determining HCV genotype. There were no significant differences between the groups in these viral factors. This study suggested that host factors induced by the HCV infection are more important than viral factors in the pathogenesis of OLP associated with hepatitis C. PMID- 9213246 TI - Wild-type p53 gene-induced morphological changes and growth suppression in hepatoma cells. AB - The human hepatocellular carcinoma (HCC) cell line, HLF, expresses only mutant type p53 (mt-p53), which has an amino acid substitution at the 244th residue from glycine to alanine. HLF cells were transfected with wild-type p53 (wt-p53) cDNA construct pC53-SN3, mt-p53 cDNA construct pC53-SCX [which differs by a single nucleotide, resulting in alanine instead of valine at the 143rd residue in p53 (p53-143)], or pCMV-Neo-Bam, as a control, by a liposome method. After G418 selection, three wt-p53 stable transformants (WT), four mt-p53 transformants (MT), and three control vector transformants (VT) were obtained. We analyzed the cell growth and morphological changes of these transformants under different culture conditions [fetal calf serum (FCS), 10%, 1%, and 0%]. Whereas no difference from control in the growth rate and morphology was observed under the 10% FCS conditions, serum starvation induced remarkable phenotypical changes in all three WTs, but not in the other transformant. Corresponding to these phenotypical changes, the transcriptional activity of wt-p53 was increased more than nine fold. These results indicated that serum starvation would induce wt-p53 biological function, which is tightly linked to morphological changes and growth suppression. To induce these changes, the introduction of the wt-p53 gene itself was not sufficient, and additional triggering, i.e., serum starvation, was indispensable. PMID- 9213247 TI - Serum concentrations of soluble HLA-class I and CD8 forms in patients with viral hepatic disorders. AB - Soluble HLA-class I and CD8 molecules were determined by sandwich ELISA in patients with viral-induced hepatic disorders. As a whole, the patients with hepatic disorders (acute hepatitis: AH; chronic hepatitis: CH; liver cirrhosis: LC; hepatocellular carcinoma: HCC) showed higher sHLA-class I and sCD8 levels than normal controls (P < 0.001). AH patients had the highest sHLA-class I levels (mean, 3513 +/- 2112 ng/ml), followed by CH (2896 +/- 1290 ng/ml), LC (2293 +/- 1266 ng/ml), and HCC (2221 +/- 1212 ng/ml) sCD8 levels wer highest in AH, followed by HCC, LC, and CH, in that order. Among histologically defined C virus positive patients, sHLA-I levels were higher in those with chronic active hepatitis (CAH) 2A (3802 +/- 1124 ng/ml) than in those with chronic persistent hepatitis (CPH; 2200 +/- 711 ng/ml; P < 0.01), the levels then decreased as the disease progressed (CAH2B, 3564 +/- 1783 ng/ml, LC, 2376 +/- 1265 ng/ml). In contrast, sCD8 values showed little difference among the disorders. sHLA-class I levels showed a positive correlation with sCD8 values both in whole patients and in patients with AH (P < 0.01), but no correlation was shown, in any patients, with biochemical parameters such as GPT and GOT. These findings, taken together, suggest that hepatic destruction is not the only cause of sHLA-class I production, but that sHLA-class I levels, together with sCD8 levels, may reflect immunological activity in hepatic disorders. PMID- 9213248 TI - Establishment of a primary culture of Echinococcus multilocularis germinal cells. AB - This study was designed to establish an in vitro primary culture of germinal cells of Echinococcus multilocularis, a parasite that causes alveolar echinococcosis of the liver (AEL). We also investigated the temperature dependency of the cultured cells. The germinal cells, which originated from a human lesion, were cultured by an original fluid-suspension method at 25 degrees C or 37 degrees C for 4 weeks. Anchorage-dependent and -independent cells were observed by light microscopy, transmission electron microscopy, and immunocytochemistry to confirm their origin. Cell number and viability were examined by immunocytochemistry and mitochondrial exclusion test. The cultured cells were also inoculated into jirds (Meriones unguiculatus) to evaluate metacestode formation. Morphology and immunocytochemistry showed that the cultured cells were typically germinal cells. The cell number declined gradually over the 4-week culture period, but viability remained at 50% at 3 weeks. These findings were not associated with either of the two culture temperatures; moreover, host-associated cells were not noted in the cultured cells at 25 degrees C. The implanted cells formed metacestodes in the jird peritoneal cavity, and their histology demonstrated mature and typical alveolar-type echinococcal cysts. We successfully established an in vitro primary culture of germinal cells. This should contribute to future studies, and, hence, a better outcome for patients with AEL. PMID- 9213249 TI - Contribution of mass screening system to resectability of hepatic lesions involving Echinococcus multilocularis. AB - The prognosis for patients with alveolar echinococcosis of the liver (AEL) is excellent when the lesion is completely resected. Early detection of the disease and subsequent resection of the lesion are thus indispensable; however, the usefulness of screening systems is now controversial. This study was designed to compare screened and non-screened patients according to stage classification and to re-evaluate the effect of screening. We studied a total of 82 patients (63 screened and 19 non-screened). The stage classification showed a significant intergroup difference (P < 0.002). The largest tumors ranged from 30 to 100 mm, and there was a significant intergroup difference (P < 0.0014). Ultrasonography showed even small lesions in the screened patients. The complete resection rate was 74.6% for the screened patients, and 21.1% for the non-screened patients, showing a significant difference (P < 0.0001). The rate of unresectable lesions was higher in the non-screened patients (32%) than in the screened patients (11%), showing a significant difference (P < 0.04). The present screening system contributes to early detection and subsequent resection of AEL, leading to a better outcome. PMID- 9213250 TI - Noninvasive determination of liver collagen content in chronic hepatitis. Multivariate regression modeling with blood chemical parameters as variables. AB - To evaluate the diagnostic utility of blood biochemical parameters in assessing degree of hepatic fibrosis, serum levels of aminoterminal type III procollagen peptide (P III P), type IV collagen (IV-collagen), 7S domain of type IV collagen (7S-collagen), and hyaluronan (HA) were measured in 54 patients with chronic viral liver disease. Liver collagen content was quantified by a computer-assisted image analysis method. Significant correlations were found between the amount of collagen in the liver and serum levels of P III P (r = 0.482; P < 0.02), IV collagen (r = 0.705; P < 0.001), 7S-collagen (r = 0.771; P < 0.001), and HA (r = 0.595; P < 0.001). To optimize diagnostic efficacy, we applied multivariate regression analysis to the combined results obtained for these blood biochemical parameters and some additional laboratory tests. The correlation coefficient obtained from the application of this model was 0.809. To measure the predictive value of the proposed model, we used it to estimate collagen content in an additional 16 patients with chronic liver disease and compared the estimated to the actual amount of collagen determined by direct measurement. The correlation coefficient was 0.937. These results suggest that application of the model would be useful for the assessment of collagen content of fibrotic liver. PMID- 9213251 TI - Radiofrequency capacitive hyperthermia for unresectable hepatic cancers. AB - Radiofrequency capacitive hyperthermia combined with intra-arterial chemotherapy or chemoembolization was performed in 45 patients with unresectable hepatic cancer. Intratumor temperature was measured in 26 (57.8%) of the 45 patients. The overall mean maximum core temperature (Tmax) was 41.3 +/- 0.3 degrees C (mean +/- SE), and the Tmax was > or = 42 degrees C in nine tumors (34.6%). The tumor temperature rise was related to the power applied, embolization with degradable starch microspheres, and obstruction of the portal vein. The overall response rate was 31.1% (14/45). The response rate was 55.6% when Tmax was > or = 42 degrees C; although this was higher than the response rate of 11.8% achieved with Tmax < 42 degrees C, the difference was not significant. There were no serious adverse effects of hyperthermia. PMID- 9213252 TI - Biosynthesis and secretion of MHC class III gene products (complement C4 and factor B) in the exocrine pancreas. AB - We recently found that complement C3 is locally synthesized and secreted into the exocrine pancreas. In the present study, we attempted to demonstrate the secretion of complement C4 and factor B in the exocrine pancreas. In five samples of pancreatic fluid, both C4 and factor B proteins were detected by enzyme-linked immunosorbent assay (ELISA). Immunoblot analysis revealed the C4 and factor B molecules in pancreatic fluid to be identical with these molecules in serum. Reverse transcriptase (RT)-polymerase chain reaction (PCR) analysis in pancreatic carcinoma cell lines suggested ductal epithelial cells to be the local production sites of these proteins in the pancreas. The secretion of C4 and factor B in ductal cell lines (PANC-1 and MIA PaCa-2) was independently regulated by interleukin (IL)-1 beta, tumor necrosis factor (TNF)-alpha, and interferon (IFN) gamma; C4 secretion was induced by IFN-gamma, whereas factor B secretion was induced by IL-1 beta, TNF-alpha, or IFN-gamma. These observations indicate that: (a) complement C4 and factor B are secreted into the exocrine pancreas, (b) ductal epithelial cells appear to be the site of C4 and factor B biosynthesis, and (c) local secretion of C4 and factor B in the pancreas is differentially regulated by IL-1 beta, TNF-alpha, and IFN-gamma. PMID- 9213253 TI - Effect of oral protease inhibitor administration on gallbladder motility in patients with mild chronic pancreatitis. AB - Oral administration of a protease inhibitor (camostat) induces pancreatic hypersecretion via hormonal and neural systems in humans. Camostat may also affect gallbladder motility via these systems. The aim of this study was to evaluate the effect of camostat on gallbladder function. Gallbladder emptying in response to caerulein administration and to egg yolk ingestion was examined ultrasonographically in 15 patients with mild chronic pancreatitis before and after 6 months of camostat treatment, and in 10 control subjects. The plasma cholecystokinin concentration after yolk ingestion was measured by radioimmunoassay. Fasting gallbladder volume and contractile function, whether stimulated by caerulein or yolk, did not differ between pancreatitis patients before camostat treatment and controls. Plasma cholecystokinin levels, basal and yolk-stimulated, did not differ between nontreated pancreatitis patients and control subjects. Fasting volume had decreased significantly by 1, 3, and 6 months of camostat treatment, while contractile function was not affected. Camostat did not influence plasma cholecystokinin levels. Oral administration of a protease inhibitor appears to decrease fasting gallbladder volume via a mechanism other than cholecystokinin release. PMID- 9213254 TI - Effect of peripherally and centrally administered calcitonin on gallbladder emptying in dogs. AB - The effect of calcitonin on meal-stimulated gallbladder emptying (GBE) was examined after intravenous (i.v.) and intracerebroventricular (i.c.v.) administration in six mongrel dogs. The gallbladder contraction was surveyed by means of real-time ultrasonography in conscious dogs. Calcitonin given i.v. elicited an immediate and strong inhibition of postprandial GBE-the integrated 0- to 120-min gallbladder response was 118.1 +/- 8.0%.h after placebo, whereas it was 91.8 +/- 2.1%.h, 59.4 +/- 17.9%.h (P < 0.001), and 14.2 +/- 20.5%.h (P < 0.001) after 3.6, 18.0, and 90.0 pmol.kg-1 calcitonin, respectively. After i.c.v. administration (1.8 and 18.0 pmol.kg-1), only the higher calcitonin dose exerted a moderate inhibitory effect on postprandial GBE. The calcitonin doses required to evoke a 50% inhibition of meal-stimulated GBE were 15- to 10-fold lower after i.v. than i.c.v. application. Peripherally given calcitonin brought about a dose dependent increase in the interdigestive gallbladder volume-the linear regression of the relative gallbladder volume versus calcitonin dose was y = 11.60 [ln(dose + 1)] + 97.02 (r = 0.864, P < 0.001). Intravenous application of calcitonin did not affect caerulein-induced GBE. The results obtained imply that: (i) calcitonin exerts an inhibitory influence on meal-induced GBE and that this effect is more pronounced after i.v. than after i.c.v. administration, and (ii) peripherally given calcitonin does not inhibit caerulein-induced gallbladder contraction in the dog. PMID- 9213255 TI - Multiple early esophageal cancers arising from Barrett's esophagus, and a review of cases of early adenocarcinoma in Barrett's esophagus in Japan. AB - A case of early esophageal adenocarcinoma arising in Barrett's esophagus is reported. Many cases of Barrett's esophagus, which is considered a premalignant condition, have been reported in Western countries, but few cases have been reported in Japan. The patient, a 53-year-old man with nausea and vomiting, was a drinker (four glasses wine/day for about 30 years), but did not smoke. He had had a hiatal hernia of the esophagus. Since endoscopic biopsies demonstrated an early adenocarcinoma in Barrett's esophagus, subtotal esophagectomy was performed. In the resected esophageal material, Barrett's esophagus was seen to extend for 12 cm. In addition to the cancer detected preoperatively as a 0-IIc lesion (1.5 cm in diameter), a 0-IIb lesion (1.5 cm in diameter) was also detected in the postoperative survey. Both lesions were well differentiated adenocarcinoma that had invaded only into the lamina propria mucosa. The 23 cases of early adenocarcinoma in Barrett's esophagus that have been reported in Japan were reviewed, and it was learned that the present case is the second of multiple early cancer arising in Barrett's esophagus so far reported in Japan. PMID- 9213256 TI - Gastric cancer associated with overexpression of parathyroid hormone-related peptide (PTHrP) and PTH/PTHrP receptor in relation to tumor progression. AB - Parathyroid hormone-related peptide (PTHrP) is involved in cell proliferation in both neoplastic and non-neoplastic tissues. We describe an autopsy case of gastric cancer in a patient who showed serum hypercalcemia and overexpression of PTHrP and PTH/PTHrP receptor in the metastatic tumor cells. The primary gastric tumor was poorly differentiated adenocarcinoma, and multiple metastases were present in the bone, multiple visceral organs, peritoneum, and lymph nodes. PTHrP and its mRNA were detected only in the metastatic tumor cells, but not in primary gastric tumor. PTH/PTHrP receptor was also demonstrated immunohistologically in metastatic tumor cells. This case suggests that the expression of PTHrP is related to tumor progression and the poor prognosis in tumors associated with humoral hypercalcemia. PMID- 9213257 TI - Three cases of familial ulcerative colitis--in a mother and two of her sons. AB - A pedigree of familial ulcerative colitis with their HLA haplotypes is reported. The mother and two children, the eldest and second of three sons were affected. The mother developed proctitis at age 35, and the lesion extended to the entire colon at the time of a relapse. The two sons developed ulcerative colitis in a similar fashion; total colitis, of moderate severity, developing at age 16 in both. The analysis of HLA haplotypes of all five family members suggested that the HLA responsible for ulcerative colitis in this family was not HLA B, C, or DR, but Aw24 and DQw1. The third son had the same HLA haplotype as the second son, and it was presumed that he would develop ulcerative colitis in the future. These cases, together with other cases of familial ulcerative colitis indicate that Aw24 and DQw1 are critical phenotypes for ulcerative colitis in Japan. PMID- 9213258 TI - Transcatheter arterial embolization-induced bilious pleuritis in a patient with hepatocellular carcinoma. AB - A 52-year-old man with hepatocellular carcinoma (HCC) was admitted with cough and fever. He had undergone four series of treatments, including transcatheter embolization and chemoembolization with lipiodol and anticancer drugs, over the previous 2 years. Computed tomography demonstrated dilated hepatic ducts, localized necrosis in the right hepatic lobe, and subphrenic abscess. He died of respiratory failure, because of increased effusion of the right pleura, about 3 weeks after admission. Autopsy revealed adhesions in the lower lobes of the right lung, diaphragm, and liver, with granulomas with bile pigment. A fistula was observed from the necrotic regions of the right hepatic lobe to the pleura through the diaphragm. A tumor thrombus in the portal trunk was histologically confirmed as well and moderately differentiated HCC with trabecular arrangement. Direct invasion of HCC with necrotic tissue to the pleura through the diaphragm appeared to have caused the respiratory failure. Although bilious pleuritis is a rare complication of transcatheter arterial embolization (TAE), it should be considered as an adverse effect of TAE in patients with a dilated hepatic duct. PMID- 9213259 TI - Prolonged intrahepatic cholestasis in acute-onset, severe autoimmune hepatitis. AB - A 72-year-old woman was admitted because of jaundice and hepatocellular dysfunction. She was diagnosed with autoimmune hepatitis from laboratory test results showing high titers of antinuclear antibodies and negativity for hepatitis viral markers. Steroid i.v. pulse therapy and oral administration of prednisolone were effective in improving the liver function test results, except for hyperbilirubinemia. Elevated serum bilirubin levels, of approximately 20 mg/dl persisted for more than 6 months, despite the administration of ursodeoxycholic acid. Insulin-glucagon therapy was given for normalization of transaminases and then withdrawn 3 weeks after admission, but it was resumed at 3 months, resulting in a dramatic decrease in serum bilirubin levels, which then normalized in 2.5 months. Liver biopsy 6 months after onset showed chronic active hepatitis with bile plugs. Insulin-glucagon therapy, because of its choleretic effect, may be worth continuing even after recovery of acute hepatic failure. PMID- 9213262 TI - Group II phospholipase A2 is not a major modifier of colonic adenomatous polyps in humans. PMID- 9213261 TI - Molecular mechanisms of hepatic fibrosis and principles of therapy. AB - Tremendous insights into the understanding of hepatic fibrosis have taken place over the past ten years. Foremost among these is the recognition that hepatic stellate cells (formerly known as lipocytes, Ito cells, or fat-storing cells) play a central role based on their ability to undergo activation following liver injury of any cause. Stellate cell activation is a broad phenotypic response, characterized by distinct functional changes in proliferation, contractility, fibrogenesis, cytokine secretion, and matrix degradation. Insights gained into the molecular regulation of hepatic stellate cell activation will lead to new, targeted approaches to hepatic fibrosis in the future, and could lead to reduced morbidity and mortality in patients with chronic liver injury. PMID- 9213260 TI - The role of apoptosis in intestinal disease. PMID- 9213263 TI - Controlled-release melatonin implants delay puberty in rats without altering melatonin rhythmicity. AB - There is increasing evidence that continuous availability of melatonin via implants can produce the same physiological changes in animals as timed administration of the hormone. The mechanisms underlying this apparent contradiction are not known. In an attempt to gain further understanding of the way continuous melatonin administration affects reproductive activity, the effects of melatonin implants on gonadal development and melatonin production were investigated in rats treated neonatally with testosterone. Five-day-old male rats maintained on a 12L:12D photoperiod were injected with 1 mg testosterone propionate to induce photo-responsiveness and implanted at 21 days of age with novel melatonin implants designed to raise the daytime blood melatonin concentration into the nighttime range, i.e., from less than 60 pM in the controls during the day to 380 +/- 33 pM in the implanted rats. Following 21 days treatment, seminal vesicle and ventral prostate weights of implanted rats were significantly less than the controls (27.0 +/- 1.9 vs. 18.5 +/- 1.5 mg/ 100 g BW (P = 0.003) and 33.8 +/- 2.1 vs. 26.7 +/- 2.2 mg/100 g BW (P = 0.02), respectively). To determine the effect of the implants upon melatonin production, urine was collected at hourly intervals during the last four days of the experiment and the hourly 6-sulphatoxymelatonin (aMT.6S) excretion rate was determined. Rats bearing melatonin implants maintained a rhythm of aMT.6S excretion in 12L:12D, which was indistinguishable from that in the control animals except for a raised daytime excretion of the metabolite. Following one cycle of urinary aMT.6S measurements in the light/dark cycle, the animals were released into constant darkness, with the implants still in place or after their removal four hours before darkness to evaluate the characteristics of the melatonin rhythm in the absence of masking effects of the light/dark cycle. The melatonin rhythm persisted in both control and implanted rats and no differences in the onset, offset, or amplitude could be determined. The results of this study indicate that, like many other mammals, for laboratory rats controlled continuous release of melatonin can mimic the effects of short daylength or timed melatonin administration. Despite the reproductive consequences of continuous melatonin delivery, the timing of endogenous melatonin production is unaffected. PMID- 9213264 TI - Cataractogenesis and lipid peroxidation in newborn rats treated with buthionine sulfoximine: preventive actions of melatonin. AB - The purpose of this study was to examine the influence of exogenously administered melatonin on cataract formation and lipid peroxidation in newborn rats treated with buthionine sulfoximine (BSO), a drug which inhibits the rate limiting enzyme in glutathione (GSH) synthesis, gamma-glutamylcysteine synthase, thereby depleting animals of their stores of the important intracellular antioxidant, GSH. BSO (3 mmol/kg BW) was given for three consecutive days beginning on postnatal day 2; melatonin (4 mg/kg) was injected daily beginning on postnatal day 2 and continuing until the animals were killed (either day 9 or day 17 after birth). None of the control animals (rats treated with neither BSO nor with melatonin) developed lenticular opacification during the observation period. In the BSO-treated rats, 16 of 18 animals (89%) had observable cataracts when they were examined. In rats that received both BSO and melatonin, the incidence of cataracts was highly significantly decreased, i.e., only 3 of 18 rats (7%) had observable cataracts. In addition to cataracts, the level of lipid peroxidation products (malondialdehyde (MDA) and 4-hydroxyalkenals (4-HDA)) was examined in the lens, brain, liver, lung, and kidney of control and experimental animals. In BSO-treated rats, the lens, kidney, and lung exhibited increased levels of MDA plus 4-HDA relative to those measured in the control rats; these increases were reversed in the BSO-treated rats who were injected with melatonin daily. While BSO administration did not increase basal levels of MDA plus 4-HDA in either the brain or liver, melatonin reduced levels of lipid peroxidation products below those measured in the control rats (at 17 days after birth). The changes induced by melatonin are consistent with the free-radical scavenging and antioxidative properties of this indole. PMID- 9213265 TI - Estrogenic effects on urinary 6-sulphatoxymelatonin excretion in the female rat. AB - The pattern of melatonin production during the estrous cycle of the rat was measured by monitoring urinary 6-sulphatoxymelatonin (aMT.6S) excretion. Adult rats were maintained under a 14L:10D photoperiod and urine was collected at hourly intervals over a 5-day period using an automated collection system; the concentration of aMT.6S was assayed by RIA and hourly outputs were calculated. Each nightly collection of urine was assigned to an estrous cycle stage as determined by the vaginal smear of the preceding morning. Total aMT.6S excretions (mean +/- SEM) during estrous, metestrous, diestrous, and proestrous stages were 493 +/- 49, 539 +/- 44, 562 +/- 40, and 646 +/- 51 pmol/night, respectively (n = 7). The excretion of aMT.6S was significantly higher on the night of proestrus compared to each of the other stages (P < 0.05). To determine whether estrogen was responsible for the increased aMT.6S excretion during proestrus, rats were studied before and after ovariectomy and following implantation with estradiol implants. Total overnight aMT.6S excretion was reduced by 31% in ovariectomized animals relative to the intact state (P < 0.05) and restored to the intact levels by administration of estradiol (P < 0.05). It was concluded that estradiol can modulate melatonin production in adult rats, and that the changing pattern of aMT.6S excretion throughout the estrous cycle may provide a basis for a functional relationship between pineal activity and reproduction in this species. PMID- 9213266 TI - A combined immunohistochemical and electron microscopic study of the second cell type in the developing sheep pineal gland. AB - Ultrastructural and immunohistochemical techniques were used to study the second cell type in sheep embryo pineal glands. Thirty-two embryos were studied from day 54 of development through birth. Specimens were arranged in four age groups, defined in terms of the most relevant histological features: Group 1 (54-67 days of prenatal development), Group 2 (71-92 days), Group 3 (98-113 days), and Group 4 (118-150 days). At 98 days, a second cell type was observed which differed from pinealoblasts and showed uniform ultrastructural characteristics similar to those of astrocytes in the central nervous system. Ultrastructural homogeneity was not matched by the results of histochemical and immunohistochemical analysis: while all Type II cells stained positive to phosphotungstic acid hematoxylin, only 50% expressed glial fibrillary acidic protein. In the course of ovine intrauterine development, the vascular affinity of this second cell population, composed of glial-like or astrocytic cells at varying stages of maturity, leads to the formation of a limiting pineal barrier. This barrier may constitute the morphological expression of a hypothetical functional involvement in the exchange of substances between blood and pineal parenchyma. PMID- 9213267 TI - Immunohistochemical and tracer studies of macrophages/microglia in the pineal gland of postnatal rats. AB - The pineal gland of rats of various ages (1-21 days old) was examined by immunohistochemistry and electron microscopy. Numerous widely distributed cells identified as macrophages/microglia were immunoreactive with the monoclonal antibodies OX-42, OX-18, OX-6, and ED1, indicating that they expressed complement type 3 (CR3) receptors, major histocompatibility complex class I and II antigens, and antigens of monocyte/macrophage lineage as detected by the antibodies, respectively. Following an intraperitoneal injection of rhodamine isothiocyanate (RhIC) in all age groups, the cells emitted a bright fluorescence. They were also labeled by horseradish peroxidase (HRP), as demonstrated in both light and electron microscopy. An HRP reaction was observed in vesicles and lysosomes at the ultrastructural level. A remarkable feature was the uptake of these tracers by pinealocytes. In light microscopy, the pinealocytes showed a punctate reaction product 3-24 hours after HRP injection. By electron microscopy, the reaction product was observed in vesicles, lysosomes, and some rod-like structures in the cytoplasm. On the basis of their immunophenotypic features, it is suggested that the macrophages/microglia in the pineal gland are active phagocytes which are also probably involved in the immunoregulatory function in the gland. The avid uptake of RhIC and HRP from the circulation by these cells suggests that serum derived substances that may gain access to the parenchyma of the gland are being constantly monitored. The labeling of pinealocytes with HRP suggests that the functional activities of these cells are being modulated by serum-derived substances. PMID- 9213268 TI - Phototransduction cascade and circadian oscillator in chicken pineal gland. AB - The chicken pineal gland has an endogenous circadian oscillator that controls the diurnal oscillation of N-acetyltransferase activity responsible for melatonin rhythm. It has been speculated that the chicken pineal cell contains a photoreceptive molecule that receives the environmental light signal and transmits the signal to the oscillator for resetting the phase. In spite of several lines of evidence suggesting the similarity between retinal and pineal photon-signal transducing proteins, the identity of the photoreceptive molecule had been an open question. In 1994, we isolated a pineal cDNA encoding a novel photoreceptive molecule and named it "pinopsin." The protein expressed in 293EBNA cells bound 11-cis-retinal to form a blue-sensitive pigment with an absorption maximum at about 470 nm. A putative G-protein interaction site of pinopsin shared a relatively high similarity in amino acid sequence to that of rhodopsin, implying that pinopsin functionally couples with transducin or transducin-like G protein(s) in the pineal cells. We have cloned a cDNA for chicken pineal transducin alpha-subunit, and the deduced amino acid sequence contained a potential site to be ADP-ribosylated by pertussis toxin (PTX). Therefore, the transducin-mediated pathway could be blocked by PTX, though previous studies showed that treatment of the cultured chicken pineal cells with PTX had no effect on the light-induced phase-shift of the oscillator. Accordingly, it is unlikely that transducin mediates the light-input pathway to the oscillator, which may involve PTX-insensitive G-protein(s) or some unidentified component(s). The G protein coupled receptor-mediated signaling processes regulating melatonin synthesis are discussed. PMID- 9213269 TI - Melatonin and N-tert-butyl-alpha-phenylnitrone block 60-Hz magnetic field-induced DNA single and double strand breaks in rat brain cells. AB - In previous research, we have found an increase in DNA single- and double-strand breaks in brain cells of rats after acute exposure (two hours) to a sinusoidal 60 Hz magnetic field. The present experiment was carried out to investigate whether treatment with melatonin and the spin-trap compound N-tert-butyl-alpha phenylnitrone (PBN) could block the effect of magnetic fields on brain cell DNA. Rats were injected with melatonin (1 mg/kg, sc) or PBN (100 mg/kg, ip) immediately before and after two hours of exposure to a 60-Hz magnetic field at an intensity of 0.5 mT. We found that both drug treatments blocked the magnetic field-induced DNA single- and double-strand breaks in brain cells, as assayed by a microgel electrophoresis method. Since melatonin and PBN are efficient free radical scavengers, these data suggest that free radicals may play a role in magnetic field-induced DNA damage. PMID- 9213270 TI - Melatonin inhibits vasospastic action of hydrogen peroxide in human umbilical artery. AB - We evaluated the antioxidant property of melatonin as it relates to the vasospastic effect of hydrogen peroxide (H2O2) on the human umbilical artery. Helical sections of umbilical arteries were obtained from healthy pregnant women who were delivered between weeks 37 and 39 of gestation. Changes in maximal potassium chloride (KCl)-induced tension were measured in arterial segments with intact endothelium. Segments were treated with H2O2 alone, or were pretreated either with an H2O2 scavenger (catalase, 2000 i.u.), a hydroxyl radical scavenger (mannitol, 10(-2) M), a nitric oxide-synthesis inhibitor (L-NG-monomethyl arginine, LNMA, 2 x 10(-4) M), or melatonin (10(-6) M to 10(-4) M). The effect of H2O2 (10(-4) M) on the relaxation induced by the calcium ionophore A23187 was also determined in arterial segments, with or without pretreatment with melatonin (10(-6) M, 10(-4) M). H2O2 (10(-6) M to 10(-4) M) potentiated vascular tension in a concentration-dependent manner (P < 0.0001). Pretreatment with LNMA significantly suppressed the vasospastic effect of H2O2 (P < 0.0001). Pretreatment with either catalase or mannitol significantly reduced the vasospastic effect of H2O2 (P < 0.005, P < 0.002, respectively). Melatonin also significantly reduced the vasospastic effect of H2O2 in a concentration-dependent manner (H2O2 10(-6) M, P < 0.0001 : H2O2 10(-5) M, P < 0.0001 : H2O2 10(-4) M, P < 0.00001). Pretreatment with H2O2 significantly inhibited the relaxation induced by the calcium ionophore A23187 (P < 0.005). Treatment with melatonin prior to exposure to H2O2 significantly restored the relaxation induced by A23187 (P < 0.005). Results suggest that H2O2 potentiates vascular tension in the human umbilical artery, perhaps by suppressing the endothelial synthesis of nitric oxide. Melatonin significantly suppressed the vasospastic effect of H2O2, possibly due to its ability to scavenge the hydroxyl radical. PMID- 9213271 TI - Melatonin in mast cells and tumor radiosensitivity. PMID- 9213272 TI - Direct spinal curvature digitization in scoliosis screening--a comparative study with Moire contourgraphy. AB - The forward bending test, widely used in scoliosis screening, is associated with high false-positive rates. We postulate that direct surface measurement of the spinal curvature by digitization of the spinous processes used in combination with the forward bending test could increase the predictive value of detecting scoliosis without sacrificing sensitivity. Sixty consecutive patients referred from a school screening program were included in this study. All had moire contourgraphy, spinous process digitization, and erect frontal radiographs of the spine. The number of false positives from the forward bending test was 35. Moire contourgraphy did not reduce the number of false positives significantly. A posterior spinal deformity of 10 degrees or more predicted scoliosis in all patients, with 15 false-positive readings. Direct measurement of the spinal curvature by spinous process digitization can be an effective second-stage screening tool for scoliosis in centers where all the screened positives are referred to a single center. PMID- 9213273 TI - Natural history of adolescent thoracolumbar and lumbar idiopathic scoliosis into adulthood. AB - Thirty-four patients with adolescent idiopathic thoracolumbar, lumbar, or lumbar components of double major curves between 20 and 55 degrees were identified. This study group was compared with an age- and sex-matched control group with regard to back pain, radicular symptoms, and perception of handicap. The objectives of this study were to define the natural history of moderate-range adolescent idiopathic thoracolumbar, lumbar, and double major curves with a lumbar component in this range. Studies that exclusively examined the natural history of thoracolumbar and more caudad curves have not been undertaken. Data from other related studies is often clouded by various factors. We reviewed the charts and radiographs of 363 patients with idiopathic scoliosis seen between 1935 and 1975 with available original radiographs. Thirty-four of 65 patients (52%) answered a questionnaire pertaining to severity of pain, functional abilities, and perceived quality of life. The same questionnaire was answered by 31 age- and sex-matched controls for comparison. The average follow-up was 22 years, and average patient age at current follow-up was 36 years. Curves at skeletal maturity measured an average of 35 degrees. On a scale of 1-10 (severe), current low-back pain in the study group was rated a mean of 3.19 versus 1.94 in the control group. Twelve of 34 patients in the study group (35%) reported no back pain, versus 21 of 31 (68%) in the control group. Twenty-four percent of the study group had radicular symptoms compared with 16% of the control group. None of the 34 study patients and 1 control patient underwent surgery for back pain. With an average follow-up of 22 years, the study group reported handicap scores comparable to those of the control group. The average age of the study patients was only 36 years, but it is encouraging that these individuals have continued to do well for at least 20 years past skeletal maturity. PMID- 9213274 TI - Correction of experimental scoliosis by rib resection in the transverse plane. AB - Experimental scoliosis with the potential for marked progression was treated by rib resection on the concave side of the curve, and the alterations of the rib cage and vertebrae in the transverse plane were investigated. Twenty-four chickens were divided into four equal groups (groups R, P, PR, and C) and pinealectomy was performed at 3 days of age in groups P and PR. In group R, three unilateral ribs were resected at the age of 4 weeks. In group PR, three ribs on the concave side of scoliosis were resected at 4 weeks of age if scoliosis of > 20 degrees developed before the age of 4 weeks. Group C served as a control. Spinal radiographs and computed tomography scans at the apical vertebrae were taken at 20 weeks of age, and spinal deformities were evaluated. Scoliosis developed markedly in groups R and P, whereas it was mild in group PR. The apical vertebrae rotated to the convex side of the curve in all groups, in the same way as it would in human idiopathic scoliosis. In group PR, the Cobb angle and the rotation angle of the apical vertebra were symmetrically suppressed. This study indicated that rib resection might control the progression of scoliosis not only in the frontal plane but also in the axial plane when it was done on the concave side of the scoliosis. Although this experiment succeeded in chickens, application in humans is uncertain. PMID- 9213275 TI - Adult spondylolisthesis treated with posterolateral lumbar fusion and pedicular instrumentation with AO DC plates. AB - Attainment of successful lumbar fusion in adults with spondylolisthesis has historically been unpredictable. Recent results and conclusions have been conflicting regarding the role of instrumentation in improving the fusion rate and clinical outcome in this patient population. In a retrospective multicenter clinical study, we assessed the outcome of 42 adults with spondylolisthesis who underwent posterolateral lumbar fusion by using pedicular instrumentation with AO DC plates. No attempt was made to reduce slippage. Follow-up clinical outcome was obtained from a patient questionnaire administered and assessed by an independent reviewer. Fusion status was assessed by anteroposterior, lateral, and oblique radiographs at the most recent follow-up examination. Spondylolisthesis was classified as degenerative in 21 patients and isthmic in 21 patients. Solid fusion was achieved in 32 (76%) patients; pseudoarthrosis occurred in two (5%) patients; the fusion mass was indeterminate in eight (19%) patients. Clinical outcome parameters rated 73% excellent to good and 27% fair to poor. Complications included four infections and two screw breakages. Poor results correlated strongly with cigarette smoking and multiple previous surgeries. In this study, fusion rate and clinical outcome were consistent with previous reports of adult spondylolisthesis. Rates of successful fusion varied according to the type of spondylolisthesis. PMID- 9213276 TI - Prevention of deep-vein thrombosis after major spinal surgery: a comparison study of external devices. AB - We studied the difference in postoperative thrombotic complications after major spinal surgery between two commonly used external compression devices. Our 136 subjects were prospectively randomized to receive either thigh-high sequential pneumatic compression wraps or pneumatic foot-compression wraps. All were studied postoperatively with duplex ultrasonography and analyzed for leg swelling, the rate of thrombotic events, and overall subjective patient comfort. The rate of postoperative thrombosis was 1.5%. The one pulmonary embolism was successfully treated with heparin. One other patient had a small asymptomatic thrombus of the saphenous vein. Both subjects wore foot wraps. Subjectively, the devices were rated as being equally comfortable. We believe that external compression for thrombosis prophylaxis after major spinal surgery is effective. The particular device chosen may be driven by other factors such as cost, physician or nursing preference, and ease of application. PMID- 9213277 TI - Treatment of coccidioidal spinal infection: experience in 16 cases. AB - Sixteen patients with spinal infection from Coccidioides immitis were treated. Lesion location was cervical in two, thoracic in four, lumbar in six, sacroiliac joint in one, and disseminated spinal in three. The neurological status was intact in 11 patients. One patient had incomplete quadriplegia, three patients had incomplete paraplegia, and a fifth patient had a lumbar root lesion. Treatment was medical only in 4 patients (one of whom required surgery 2 years later) and combined medical and surgical in 13 patients. All patients received amphotericin B intravenously. Follow-up averaged 24 months in 15 patients (range, 12-42 months). The outcome in four patients treated medically alone was one death, one remission, one relapse with disease progression, and one without follow-up. The outcome in the combined medical and surgical group was nine fusions, one pseudarthrosis, and three lesional excisions, all with remission. Successful treatment outcome is disease arrest, as opposed to "cure." PMID- 9213278 TI - Cervical spine manipulation: summary report of a systematic review of the literature and a multidisciplinary expert panel. PMID- 9213279 TI - An in vivo analysis of the dimensional changes of the neuroforamen after anterior cervical diskectomy and fusion: a radiologic investigation. AB - Eighteen patients (11 men, 7 women; average age, 45.5 years) who underwent anterior cervical diskectomy and fusion (ACDF) for the treatment of radiculopathy had preoperative and immediate postoperative computed tomography (CT) scans to measure pre- and postoperative foraminal heights and foraminal areas, preoperative disk space height, and postoperative graft height. The mean foraminal height preoperatively was 0.851 cm; postoperatively, it was 1.01 cm, with a mean percentage increase of 20% (-8.8 to 56.8%). Mean preoperative foraminal area was 37.53 mm2, increasing to a mean of 49.04 mm2 postoperatively with a mean percentage increase of 33% (range, -1.5 to 76.9%). No significant correlations between graft height and change in maximal foraminal height or foraminal area or between changes in foraminal height or area and postoperative symptom relief were found. Although significant increases in foraminal dimensions were seen radiographically after ACDF, these increases were variable and not strongly related to graft height. In addition, the increases in foraminal dimensions were not related to the short-term clinical results of ACDF. This study fails to support the hypothesis that the reliable results of ACDF can be ascribed primarily to indirect decompression of the uncovertebral foramen by disk space distraction. PMID- 9213281 TI - Screw omission and the stability of posterior pedicle screw constructs for short segment stabilization. AB - To determine the net contribution of a spinal construct to stability, and whether extending the construct to another level in situations in which a defective pedicle cannot have a screw inserted, we performed biomechanical tests in which we evaluated three-, four-, and five-level synthetic spinal constructs in which the location and number of pedicle screws were varied above and below a vertebrectomy defect. We subjected all constructs to axial, compression, lateral bending, flexion, extension, and torsional forces with the use of an Instron biaxial machine. Left-right symmetrical constructs were more stable than asymmetrical ones. Three-level constructs were statistically stiffer than the longer ones in compression, left bending, and flexion. Torsional stability, however, was greater in the longer constructs. Five-level constructs with both end screws in place had greater torsional stiffness than when they were missing a screw. In vertebrectomy defects, if four screws cannot be placed across it, then the engagement of two screws is indicated. The stability provided by a single screw at a spinal level is minimal. Additional screws augment the purchase of the construct in the bone; however, they do not afford further protection to the defect. PMID- 9213280 TI - Radiologic evaluation of the relation of the screw tip to the nerve root in the intervertebral foramen. AB - Eight cervical spines were used to evaluate the relation of the screw tip to the nerve root in the intervertebral foramen. The specimens were divided into two groups: (a) lateral placement without contact with the nerve root, and (b) lateral placement with penetration of the nerve root. Six screws were used for each specimen. After screw placement, oblique radiographs and axial computed tomography (CT) scans were taken. The results on oblique radiographs showed that 23 (95.8%) of 24 screws without contact with the nerve root were found in the upper zone or the junction between the upper and lower zones of the intervertebral foramen. Twenty (83.3%) of 24 screws with penetration of the nerve root were located in the junction between the lower zone of the intervertebral foramen and the pedicle zone. No definite diagnosis of screw penetration of the nerve root could be made on axial CT scans, although scans can show that the screw is violating the foramen. Whether or not a screw violating the intertransverse foramen and affects the nerve root depends on its position on the oblique radiograph. It may be not necessary to remove or change the screw immediately if a longer screw is found in the upper portion of the intervertebral foramen on the oblique view and angled laterally on axial CT scan in a patient without radicular symptoms. PMID- 9213282 TI - Comparison of neurologic deficits with atlanto-dens intervals in patients with Down syndrome. AB - Eighty-four patients with Down syndrome had flexion-extension lateral roentgenograms of the C1-C2 articulation for the purpose of dividing the group into subluxators (> or = 4 mm atlanto-dens interval and 2 mm translation) and nonsubluxators (those who did not meet these criteria). Neurologic examinations and chart review were carried out on all patients to ascertain those with a positive neurologic finding or history. Seventeen (20%) were defined as subluxators and 67 (80%) as nonsubluxators. Five (29%) of the subluxators were found to have positive neurologic findings and 18 (27%) of the nonsubluxators had similar types of positive neurologic findings. These percentages are not significantly different. This led us to conclude that positive neurologic findings and an abnormal atlanto-dens interval are not alone adequate criteria to judge need for surgical stabilization of the C1-C2 articulation in patients with Down syndrome. PMID- 9213283 TI - Rigid intrasegmental fixation for repair of a pars defect in a young athlete: case report and description of technique. AB - Stabilization for the treatment of a pars defect frequently involves fusion with sacrifice of a motion segment. Intrasegmental stabilization has been described, however, with preservation of the motion segment by using various constructs. We describe a method of obtaining rigid fixation across a pars defect without sacrificing a motion segment. PMID- 9213284 TI - Spontaneous correction of a traumatic kyphosis after posterior spinal fusion in an infant. AB - Posterior spinal fusion of presumed traumatic kyphosis at T11-T12 in a 23-month old child resulted in spontaneous correction of a major angular deformity. Considerable resorption of the T12 body occurred. The remodeling process was largely completed by 8 years and 8 months of age. PMID- 9213285 TI - The lumbar spine--small is beautiful: the Third Annual Ian Macnab Memorial Lecture. PMID- 9213286 TI - Almost a revolution: an international perspective on the law of involuntary commitment. AB - To what extent have developments in commitment law around the world paralleled trends in the United States in the last three decades? Although the American emphasis on dangerousness criteria and strigent procedural rights has been echoed in a number of other countries, it has not dominated reform in most nations. The leading alternative has been the 1983 Mental Health Act in England and Wales, with its focus on the "health and safety" of the patient, as well as protection of other persons, and its avoidance of judicial hearings. How have these reforms fared? Extensive data from the United States, and more limited data from other countries, suggest that reforms in general are resisted when they are seen as shifting the focus away from patients' treatment needs. When law fails to reflect widely held moral sentiments, it is molded in practice to conform more closely to those sentiments. It is helpful to recognize that a variety of approaches to mental health law are consistent with reasonable protection of civil liberties in a democratic society. Greater attention to practices in other countries may help reformers expand the menu of options in policy debates. PMID- 9213287 TI - Description of an outpatient psychiatric population in a youthful offender's prison. AB - Prisons are receiving increased numbers of inmates with mental and emotional problems. This study describes some of the characteristics and treatment of such an outpatient population. It was determined that a typical patient is a white male, 19 years old, of average intelligence, with a sporadic work record and poor academic performance, who quits high school in his freshman year. He has a history of substance abuse and is likely to have a multidrug habit. He is likely to have had a traumatic childhood and had psychiatric treatment as a child or young adolescent, as well as having attended special classes in school and counseling for drug abuse. The great majority of patients were diagnosed as having either mood, adjustment, or psychotic disorders. All were treated with a psychotropic medication and case management and also with some type of accepted individual and/or group counseling. In this population, there is a high incidence of expression of aggression requiring medication and counseling with the patient's permission. Patients responded well to treatment, but usually requested to discontinue treatment when symptoms diminished. However, approximately half of them returned for medication when symptoms recurred. PMID- 9213288 TI - Not guilty by reason of insanity of murder: clinical and neuropsychological characteristics. AB - We examined archivally clinical status, neuropsychological functioning, and perpetrator-victim relationships of 28 adult patients who had committed homicide and had been subsequently involuntarily committed to a forensic hospital. We divided patients into two groups: (1) not guilty by reason of insanity (NGRI) acquittees (n = 13) and (2) convicted murderers (n = 15). In comparison with convicted murderers, NGRI acquittees were more likely to be seen as psychotic at the time of the index offense and also were more likely to have killed blood relatives, especially a parent. By contrast, convicted murderers were more likely to have killed a significant other, mainly a spouse or lover. At the time of the index offense, substance abuse was more likely to have occurred in the convicted murderers than in the NGRI acquittees. NGRI acquittees and convicted murderers did not differ on neuropsychological tests, with both groups generally scoring within normal limits on all tests. Taken together, these results suggested that NGRI murderers may be driven by acute psychosis directed toward blood relatives and occurring against a backdrop of relatively preserved neuropsychological functioning. PMID- 9213290 TI - Detection of malingering: validation of the Structured Inventory of Malingered Symptomatology (SIMS). AB - This article discusses the development and validation of a paper and pencil screening measure, the Structured inventory of Malingered Symptomatology (SIMS), designed to detect malingering. Test items were constructed from a combination of revised validity questions from existing instruments and characteristics of malingerers noted by existing research. Items were organized on one of five subscales by experienced clinical psychologists. College students (N = 476) were assigned to one of seven simulation conditions (i.e., psychosis, amnesia, neurologic impairment, mania, depression, low intelligence, and "fake bad") or an honestly responding group. All subjects were administered the SIMS, the F and K scales of the MMPI, 16PF Faking Bad scale, and portions of the malingering scale. The SIMS total score demonstrated the highest sensitivity rating (95.6%) for detection when compared with the other validity indices. Suggestions concerning further research using the SIMS as well as its potential utility in a complete evaluation process are discussed. PMID- 9213289 TI - Aggression and schizophrenia: efficacy of risperidone. AB - The advent of novel antipsychotic medications has raised treatment expectations for patients with severe mental illness. In this regard, clozapine has been particularly effective in reducing aggressive behavior in patients with schizophrenia. This study compared the efficacy of risperidone and conventional antipsychotic medications in the management of hostile patients. Improvements in the level of aggression were evident over time in both treatment groups, and a similar response between risperidone and typical antipsychotics was observed. Future studies should address the relative role of typical antipsychotics, adjunctive agents, and novel antipsychotic medications in the pharmacological management of persistent aggression in patients with schizophrenia. PMID- 9213292 TI - Accommodations for test anxiety under ADA? AB - Test anxiety prevents students from demonstrating their knowledge on examinations. To be covered by the Americans with Disabilities Act, test anxiety must pass two legal tests. First, it must be a "mental impairment." As a form of Social Phobia, a mental disorder included in the Diagnostic and Statistical Manual of Mental Disorders, it meets this first test. Second, it must "substantially limit one or more of the major life activities." Individuals for whom test anxiety is one manifestation of Social Phobia-Generalized are substantially limited in the major life activities of interacting with others and working. Individuals for whom test anxiety is the only manifestation of their Social Phobia are substantially limited in the major life activities of thinking and working, the latter because they are excluded from any career requiring a test for application, credentialing, licensure, or training. Accommodations may include taking the test in a separate room or taking an untimed examination. Documentation supporting a diagnosis of test anxiety should include evidence of significant impairment in test performance. PMID- 9213291 TI - Competency to consent to hospitalization in the medical patient. AB - A slightly modified version (the CQ-Med) of a 15-item competency questionnaire (the CQ) was used to assess competency to consent to hospitalization in general hospital patients. The purpose of the study was to determine whether voluntary psychiatric inpatients would score comparably with general hospital inpatients using a similar questionnaire. The patients studied performed better in nearly all areas of competency than the previously studied adult and adolescent psychiatric subjects using the same questionnaire (modified for the respective study populations). The CQ-Med questionnaire may be a useful instrument for preliminary screening of general hospital patients, when indicated, for assessment of competency to consent to hospitalization. PMID- 9213293 TI - Criminal recidivism and family histories of schizophrenic and nonschizophrenic fire setters: comorbid alcohol dependence in schizophrenic fire setters. AB - Life-time criminality, family history, and situational factors during a fire setting offense were compared between 44 fire setters who had been diagnosed with schizophrenia or delusional psychosis and 260 nonpsychotic fire setters who had undergone a pretrial forensic psychiatric evaluation. The same comparisons were made between alcoholic (n = 25) and nonalcoholic (n = 19) schizophrenic fire setters. Medical and criminal records were studied. Life-time criminal histories of schizophrenic and nonschizophrenic fire setters were not significantly different with respect to multiple fire setting and violent offenses. Nonschizophrenic and alcoholic schizophrenic fire setters had, in general, a high rate of criminal offenses. The family history of schizophrenic fire setters was often characterized by the father's alcoholism and the mother's psychosis. Comorbid familial alcoholism increased life-time criminal recidivism among schizophrenic fire setters. PMID- 9213294 TI - Ethical aspects of competence for sexual relationships: a case of adult sibling incest. AB - A case of adult sibling incest serves as the occasion to review relevant literature and discuss issues related to competence to consent to sexual relations between adults. The case was presented at ethics rounds at the Massachusetts Mental Health Center (Boston, MA), and the resulting discussion is analyzed as to the decision making with which the treatment team is faced. The relationships among competence and competence assessment, autonomy, morality, and countertransference issues are also considered in this provocative case. PMID- 9213295 TI - Pretreatment minimal staging and prognostic factors for non-small cell lung cancer. PMID- 9213296 TI - Staging classification committee. PMID- 9213297 TI - Induction treatment for resectable non-small-cell lung cancer. PMID- 9213298 TI - Small cell lung cancer. PMID- 9213299 TI - Post-operative adjuvant therapy for non-small-cell lung cancer. PMID- 9213300 TI - Concurrent treatments and induction treatments for unresectable tumors. PMID- 9213301 TI - Role of positron emission tomography in the staging of lung cancer. AB - Early detection and staging of lung cancer is important in initiating rapid treatment and improving prognosis. Computed tomography (CT) and magnetic resonance (MR) imaging have a high resolution and are able to reveal structural abnormalities, but still have problems differentiating benign from malignant lesions. Lesion size is used as a distinguishing parameter but definite diagnosis still relies on invasive procedures. Positron emission tomography (PET) is based on imaging of biochemical processes in vivo. PET is unique by disclosing metabolic differences between benign and malignant disease, e.g. glucose utilization. Here, the role of PET in diagnosis and (re)-staging of lung cancer as well as monitoring of therapy response will be reviewed. PMID- 9213302 TI - Non-small cell lung cancer: clinical value of new biological predictors. PMID- 9213304 TI - Treatment of small cell lung cancer: the state of the art. AB - Many innovations have been tested to improve the power of chemotherapy for SCLC including: drug diversity enhancement, dose and dose-intensity escalation, incorporation of new agents, and resistance modification. Although superiority of combination chemotherapy over monotherapy has been shown, clinical trials have failed to demonstrate a clearly superior multiagent regimen. When dose and dose intensity are diminished from standard levels, the effect is detrimental for both limited and extensive stage SCLC. Trials of dose and dose-intensity above standard levels have not yet shown advantages for patients with extensive stage SCLC. However, the only two randomized trials of chemotherapy dose escalation for limited SCLC show statistically significant survival benefits. The optimal intensity of chemotherapy for limited SCLC has not been defined. State-of-the-art chemotherapy for extensive SCLC could be CAV, EP, or CAV/EP but EP has generally been favored because it is associated with less myelotoxicity and four cycles are considered adequate rather than 6 cycles of the others. EP is widely regarded as state-of-the-art chemotherapy for limited SCLC particularly because this regimen can be integrated with concurrent thoracic irradiation with acceptable toxicity. Early thoracic irradiation with concurrent EP chemotherapy is state-of-the-art treatment for limited SCLC, however, it must be conceded that EP has never been shown to be superior to any other chemotherapy regimen in a phase III trial of either limited or extensive SCLC. Current state-of-the-art treatment for limited SCLC can result in actual 5-year survival rates of at least 20%; declaration of a statistically significant improvement will require sample sizes than most clinical trials performed to date. PMID- 9213303 TI - Recursive partitioning analysis of 1592 patients on four Radiation Therapy Oncology Group studies in inoperable non-small cell lung cancer. AB - Survival outcome of 1592 analyzable patients on four Radiation Therapy Oncology Group (RTOG) studies in inoperable non-small cell lung cancer were studied utilizing a recursive partitioning analysis (RPA). This approach creates a regression tree according to prognostic variables which partitions into homogenous subsets according to survival. Four protocols, RTOG 83-11, 83-21, 84 03 and 84-07 were analyzed. 83-11 and 84-07 were studies utilizing irradiation with alterfractionation; 83-21 and 84-03 were studies evaluating thymocin with irradiation and prophylactic cranial irradiation with thoracic irradiation respectively. Nine pretreatment variables and one treatment variable were analyzed. Adjustment for radiotherapy effect was made in the accelerated treatment protocol (84-07). Overall, median survival for the entire group was 9.0 months with 17% alive at 2 years. Univariate analysis suggests that KPS, < or = 70 vs. 80-100, pleural effusion, weight loss, < or = 5% vs. 5%, age, 60+ vs. < 60, T stage (T1 and T2 vs. T3 and T4) and N stage (N- vs. N+) were important prognastic factors. Radiation dose, sex, race and histology were not univariate prognastic factors. RPA identified KPS as the most significant covariate (median survival 5.9 mos. < or = 70 vs. 9.9 mos. 80-100). Within KPS 80-100 other splits occurred for N stage, age, weight loss and radiation therapy dose. KPS < or = 70 split at pleural effusion only. The best overall RPA tree has four distinct classes with median survival times ranging from 3.3 to 12.6 months. The RPA classes were validated in an independent non-small cell lung cancer dataset. This analysis may allow more intelligent stratification and study-design for future RTOG trials in inoperable non-small cell lung cancer. PMID- 9213306 TI - The limits of surgical resection alone for non-small cell lung cancer. AB - Surgical resection remains the best treatment for Stages I and II non-small cell lung cancer. In Stage IIIA disease the use of induction therapy has become widespread, although evidence supporting this approach is still preliminary. However, in subsets of patients with T3 tumours without mediastinal nodal involvement and those with certain single station, non-bulky N2 disease, surgery alone is still the preferred therapy. Studies show survival rates with surgery alone the same or higher than those achieved by most induction therapy regimens. PMID- 9213305 TI - Surgery for small cell lung cancer. AB - Operative management of small cell lung cancer generally yields little benefit because these tumors are known for their propensity to disseminate early to regional lymph nodes and distant sites. Primary surgery followed by chemotherapy is however indicated in very early stage tumors where survival approximates that of resected non small cell lung tumors. Surgery as an adjuvant to combination chemotherapy is also advocated by some authors to downstage the tumor and render it resectable. Candidates for salvage procedures include patients who have achieved complete response with chemotherapy, but subsequently relapsed within the chest at the site of the primary tumor. PMID- 9213307 TI - Adjuvant chemotherapy for non-small cell lung cancer. PMID- 9213308 TI - Intensity of neoadjuvant therapy in resectable non-small cell lung cancer. AB - Induction chemotherapy prior to definitive local treatment improves the survival of patients with Stage III non-small cell lung cancer. Several strategies to improve on the impact of 2 or 3 cycles of conventional induction chemotherapy are under study. Immediate concurrent chemoradiation followed by surgery has been evaluated extensively in the Phase II setting. This approach is now the subject of a Phase III trial in North America. Increasing the number of induction chemotherapy cycles or using high dose chemotherapy with autologous stem cell support are other strategies currently under evaluation. Understanding whether either of these approaches will improve survival for patients with Stage III NSCLC must await future Phase III trials. PMID- 9213309 TI - Alternatives to chemotherapy and radiotherapy as adjuvant treatment for lung cancer. AB - Because adjuvant chemotherapy has resulted in only modest prolongation of survival for patients with lung cancer, investigators have turned to the evaluation of alternative treatment strategies for this patient population. Immunotherapy with Bacillus Calmette Guerin, Corynebacterium parvum, and levamisole has been evaluated in several prospective randomized trials, and no study has shown a statistically significant difference in overall survival. Interferon has been evaluated in three trials of adjuvant therapy after response to chemotherapy for small cell lung cancer. Different interferon preparations were used, but none of the trials showed a significant prolongation of survival. The retinoids have been evaluated as adjuvant treatment after complete resection of stage IN-SCLC. One trial showed a reduction in second primary tumors, and in particular, tumors to tobacco smoking in patients treated with retinyl palmitate. A second trial using 13-cis retinoic acid is ongoing in North America. In the last decade, several inhibitors of angiogenesis have been identified, and they are now beginning to be evaluated in the clinical setting. The National Cancer Institute of Canada Clinical Trials Group and the European Organization for Research and Treatment of Cancer have initiated a study of adjuvant marimastat, a metalloproteinase inhibitor, for patients who have responded to induction chemotherapy for small cell lung cancer. This is the first adjuvant antiangiogenesis factor trial to be initiated for any tumor type. Other investigational agents which are currently undergoing Phase I and Phase II testing include monoclonal antibodies which may inhibit tumour cell growth by binding to growth factors, or which may be conjugated to toxins or chemotherapeutic agents which result in tumour cell death. In the last decade, we have witnessed an explosion in our knowledge and understanding of the regulation of normal and neoplastic cell growth at the molecular level. It remains only speculative at this time as to whether manipulation of abnormal genes in malignant cells will be clinically possible, and whether treatment of this sort may be applied in an adjuvant setting. PMID- 9213310 TI - Thoracic irradiation of limited small cell lung cancer: have we defined optimal dose, time, and fractionation? AB - There is a general agreement that patients with limited small cell lung cancer (L SCLC) are better treated with a combination of thoracic radiation therapy (TRT) and systemic chemotherapy (CT) than with either modality alone. There is little agreement, however, and few data from prospective trials, to define optimal ways of combining these modalities. This article will review available data on radiation dose, timing, fractionation, and volume issues in an attempt to develop standards for current practice and suggest appropriate questions for future prospective clinical trials. PMID- 9213311 TI - [Mode of spreading and biological behavior in bile duct carcinoma]. AB - There are three macroscopic types of hepatic bile duct carcinoma, such as papillary (P-), nodular (N-) and diffuse (D-) type. Immunohistochemical studies demonstrated that P-type expressed cadherin and catenin higher than N- and D types. The expressions of both cadherin and catenin were found stronger in pap and tub1 than tub2. The nuclear area of cancer cell, correlated with both labeling index of Ki-67 and aberrant accumulation of p53, was significantly larger in the subserosal layer than in the mucosal layer. These may explain the differences in the biological behavior between P- and N, D-types. P-type grows within the mucosal layer, while N- and d-type are more invasive, developing into the subserosal layer. Our clinical data also demonstrates the poor prognosis of N , D-type of hepatic bile duct carcinoma. On these basis of the biological malignancy of N, D-type, it is critical to remain the surgical margin free from cancer cells to cure this type of hepatic bile duct carcinoma. PMID- 9213312 TI - [Chemotherapy for primary bile duct cancer]. AB - The prognosis for patients with bile duct cancer (BDC) is still dismal, because of the high incidence of postresection recurrence and frequency of unresectable tumors. Chemotherapy has been applied over 20 years with anticipation of enhanced response rate together with minimal adverse effects and prolonged survival. Despite the scarcity of experiences with chemotherapy dedicated to BDC, some efficacy of chemotherapy on BDC has been indicated by this review of the literature. Chiefly, the administered drugs to unresectable BDC were 5 fluorouracil (5-FU), adriamycin (ADM) and mitomycin-C (MMC). Systemic chemotherapy, employing those drugs as single agents or as a combination like FAM (5-FU + ADM + MMC), was most commonly administered. The collective response rate was 18.6% (13/70), including a higher response rate of 29% (4/14) by FAM. Regional chemotherapy by hepatic arterial infusion has been expected to be more effective from the view point of blood supply, however, only a few trials have been reported as long as BDC. The series from the literature show a collective response rate of 50% (3/6) for hepatic arterial infusion therapy. The total number of patients (n = 16) treated by intra-ductal chemotherapy is too small to estimate clinical efficacy despite a relatively high collective response rate of 43.8% (7/16). This paper should encourage further investigation of chemotherapy for BDC. PMID- 9213313 TI - [Intrahepatic bile duct carcinoma (cholangiocarcinoma)]. AB - There is no high risk group for cholangiocarcinoma as there is for hepatocellular carcinoma, and it has a poor prognosis because many cases are diagnosed after it has become advanced. To date, there is no effective chemotherapy or radiation therapy for cholangiocarcinoma, and extended hepatectomy is the only effective treatment. In Japan, regional lymph node dissection and extended hepatectomy have been performed aiming at curative resection, but the 5-year survival in Japan is still low, only 26.1%. The Committee on the Japanese General Rules for the Clinical and Pathological Study of Primary Liver Cancer has divided macroscopic type into 3 patterns: mass-forming type, periductal infiltrating type, and intraductal growth type, to access prognosis on a common basis. According to these groups, our patients with the intraductal growth type had a good outcome, but patients with the mass-forming type and periductal infiltrating type had a poor outcome. Many papers have reported that the presence of lymph node metastasis makes the prognosis poor. Among our cases, the 5-year survival rate for all patients who underwent hepatectomy was 26.1% and the rate for patients positive for lymph node metastasis was 10.8%, as opposed to 45.1% for patients negative for lymph node metastasis. We examined the outcome according to histological type and found that based on the histological findings, the prognosis was increasingly poor in the following order: papillary adenocarcinoma, macrotubular carcinoma, microtubular carcinoma. To achieve curative treatment in the future it will be important to clearly define the extent of hepatectomy and determine the extent of lymph node dissection required, and clearly identify other prognostic factors. PMID- 9213314 TI - [Surgical treatment of hilar cholangiocarcinoma]. AB - From the therapeutic and diagnostic viewpoints, percutaneous transhepatic biliary drainage (PTBD) is crucial for the preoperative management of hilar cholangiocarcinoma. Pertinent multiple catheterizations using PTBD produce effective relief of jaundice and accurate diagnosis of cancer extent. Endoscopic retrograde biliary drainage is contraindicated for preoperative biliary decompression. To reduce posthepatectomy liver failure, an accurate preoperative assessment of hepatic functional reserve is essential. Indocyanine green test has been used conventionally. Although this test underestimates liver function under conditions of jaundice, it is still the most practical and reliable. Preoperative portal vein embolization is an effective method for preventing posthepatectomy liver failure and extending an indication of extensive liver resection. Liver resection for hilar cholangiocarcinoma is now popular in Japan, and combined en bloc resection of the caudate lobe has become general. However, the procedure of and indication for hepatectomy is not yet standardized. Further investigations are needed to produce more rational surgical procedure for hilar cholangiocarcinoma. PMID- 9213315 TI - [Surgical treatment for the upper and middle bile duct cancer]. AB - A total of 45 patients with bile duct cancer, 13 patients predominantly with Bs bile duct cancer, 14 with Bm cancer, and 18 with Bim cancer, were chosen from 104 patients with bile duct cancer, not including gallbladder cancer or cancer of the papilla of Vater, who underwent surgical resection in our department between Sep 1974 and Nov 1996, and were evaluated with respect to surgical procedure, pathological findings, and outcome. The patients with Bs cancer were compared with 39 patients with cancer of the main hepatic duct junction who underwent hepatectomy, and the patients with Bim cancer were compared with another 20 patients with Bi bile duct cancer. The effect of combined resection of the vessels and hepatopancreatoduodectomy (HPD) on patients with advanced cancer was assessed. To attain a hw (-) margin, hepatectomy should be performed in patients with Bs bile duct cancer, unless localized. Bm and Bim bile duct cancer patients with negative margins had a relatively favorable outcome irrespective of surgical procedure. Resection of the vessels and HPD were useful in improving the resectability rate, but not in attaining longer survival. PMID- 9213316 TI - [The study for the surgical treatment on distal bile duct carcinoma]. AB - The surgical result in 74 patients with resected distal bile duct carcinoma was reviewed to clarify the surgical strategy of distal bile duct carcinoma. The clinicopathological record was according to the General rules for surgical and pathological studies on cancer of biliary tract(the 3rd edition, edited by the Japanese society of biliary surgery). 58 patients underwent standard pancreatoduodenectomy and 16 patients had pylorus-preserving pancreatoduodenectomy. The curative resection was performed in 38 patients (51.3%). The overall 5-year survival rate (operated death included) was 35.8%. The 5-year survival rate in curative resection was 56.6%. The survival rate of patients with curative resection was significantly better than that of the patients with relative non-curative (p < 0.05) or absolute non-curative resection (p < 0.01). We concluded that the Long-term survival after surgical resection was mostly correlated with curability. To obtain curative resection, the free surgical margin of hw and ew was essential. As for free "hw", intraoperative frozen dissection was indispensable. For free "ew", the dissection of the soft tissues in retroperitoneum at the back of pancreas head was necessary. On the lymph node dissection, the lymph nodes of No 8, 12, 13, 14 should be removed. PMID- 9213317 TI - [Evaluation of treatment for carcinoma of the papilla of Vater]. AB - 105 cases of resected carcinoma of the papilla of Vater were studied and 5-year cumulative survival rate of PD was 56.5% and that of each stage was as follows: stage I 93.3%, stage II 70.2%, stage III 7.3%, stage IV 0%. The most significant histological factor influencing on prognosis was pancreatic invasion. 3-year survival rate of panc1a was 75.0%, significantly much better than that of panc1b (29.8%). The lymph node metastasis was frequently noted at no. 14 (11.5%). All of no. 14 lymph-adenectomy and no. 16 lymph-adenectomy on the case suspected as pancreatic parenchymal invasion, produced long-term survive even with metastasis of no. 14. This method led improvement of prognosis of stage II and stage III determined by lymph node factor. However, each prognosis of panc 1b, 2, 3 cases was not improved by this method, especially panc2, 3 cases survived fewer than only 2 years. Extended lymphadenectomy and multimodality therapy were needed on the cases which were suspected as pancreatic parenchymal invasion. PMID- 9213318 TI - [From vascular surgery to gastrointestinal surgery]. AB - From the basic and clinical study of the peripheral vascular disturbance, my interest has gradually focused on the visceral circulation, following the first experience of an acute bowel gangrene. Thereafter, I have investigated acute mesenteric ischemia and ischemic enterocolitis both clinically and experimentally. Furthermore, I have also noticed on the combined vascular resection for the malignancies of the gastrointestinal organs. Finally, I have discussed on the role of newly introduced vascular intervention for the gastrointestinal disorders. Further development in many fields will surely lead to great possibility in the future. PMID- 9213319 TI - [Fulminant hepatic failure and liver transplantation]. AB - Fulminant hepatic failure (FHF) is defined as the onset of grade II hepatic encephalopathy within 8 weeks after the onset of jaundice in patients whose prothrombin time is less than 40%. There are an estimated 3,700 cases of FHF in Japan. There are no specific therapies for FHF, however, liver transplantation is recommended for situations in which spontaneous recovery appears unlikely. The 1 year graft survival after liver transplantation for FHF is 65 to 80%. At present, orthotopic liver transplantation has become an accepted procedure for FHF. The advantages of auxiliary transplantation in FHF are the temporary requirement for immunosuppression drugs until the host liver recovers and the relative ease of surgery. Two types of auxiliary liver support are possible: heterotopic (APL) and orthotopic (APOLT). APLT is technically easier but suffers from competition between the graft and native liver for portal blood supply, problems of venous congestion and potential lack of space. However, APOLT has a more secure portal blood supply than APLT. Several technical problems with auxiliary transplantation need further evaluation. PMID- 9213320 TI - [A successful surgical treatment of dissecting thoracoabdominal aortic aneurysm using a heparin coated percutaneous cardiopulmonary support system]. AB - A 52-year-old man with dissecting thoracoabdominal aortic aneurysm (impending rupture) involving the celiac, superior mesenteric and renal arteries underwent graft replacement of the thoracoabdominal aorta with reconstruction of all visceral branches and intercostal arteries (Th11,12) with the aid of femoro femoral bypass. The bypass was performed using a heparin coated percutaneous cardiopulmonary support system with low doses of heparin, maintaining activated coagulation time at about 300 seconds. During the reconstruction of the major visceral branches, the branches were separately cannulated from inside the aorta, and perfused selectively via partial extracorporeal circulation. The intercostal arteries were reconstructed segmentally to minimize the duration of ischemia. During the repair of the intercostal arteries, Fogarty balloon catheters were inserted into the intercostal arteries to prevent back bleeding and ischemia. The patient had a satisfactory postoperative course. PMID- 9213321 TI - Docetaxel in breast cancer and a rationale for combination therapy. AB - Development of the taxoids has progressed rapidly in the 1990s. In vitro studies have demonstrated that docetaxel (Taxotere) has a longer residence time and higher accumulation within tumor cells than paclitaxel (Taxol), possibly accounting for its greater cytotoxicity. Animal studies have shown docetaxel to possess high antitumor activity. In clinical studies, docetaxel as a single agent has been shown to be highly active against a variety of solid tumors. It is at least as active in metastatic disease as other agents currently used, and it has demonstrated effectiveness against tumors resistant to anthracyclines or paclitaxel. Docetaxel shows some activity in cell lines resistant to fluorouracil (5-FU), vincristine, cisplatin (Platinol), and etoposide (VePesid). Its unique mechanism of action and lack of cross-resistance or overlapping toxicities with many agents may also contribute to its efficacy and safety in combination therapy. PMID- 9213322 TI - Docetaxel in combination with doxorubicin: a phase I dose-finding study. AB - This phase I dose-finding study examined the effects of the combination of doxorubicin and docetaxel (Taxotere) in 42 women with metastatic breast cancer. The combination was studied at six different dosing levels. The maximum tolerated doses were defined as doxorubicin, 50 mg/m2, and docetaxel, 85 mg/m2, with sepsis as the dose-limiting toxicity. Activity was observed at all dose levels, especially at the highest dose levels (50/60, 50/75, 50/85, and 60/60 mg/m2), with a response rate of 81% (95%; confidence interval, 62.5% to 92.5%) in patients treated at these dose levels. The response rate in patients with visceral disease was 74%, and 82% in patients with liver metastasis. Adjuvant chemotherapy with or without anthracyclines did not affect the response rate. The recommended doses were doxorubicin, 50 mg/m2, and docetaxel, 75 mg/m2, or 60 mg/m2 of both drugs, administered on day 1 every 3 weeks, without granulocyte colony stimulating factor (G-CSF, filgrastim [Neupogen]) support. Neutropenia was the only grade 3 or 4 adverse event. There were no cases of congestive heart failure, a significant decrease in left-ventricular ejection fraction, or interruption of treatment because of fluid retention. PMID- 9213323 TI - Docetaxel and cyclophosphamide in patients with advanced solid tumors. AB - This trial was designed to determine the recommended maximum tolerated dose (MTD), toxicity, pharmacokinetics, and efficacy of docetaxel (Taxotere) and cyclophosphamide (Cytoxan, Neosar) for phase II studies. Both drugs were administered to 39 patients with advanced solid tumors, 26 of whom had breast cancer. Docetaxel doses ranged from 60 to 85 mg/m2 and cyclophosphamide doses ranged from 600 to 800 mg/m2. All patients received steroid prophylaxis. The MTDs for patients with a history of prior chemotherapy were 75 mg/m2 of docetaxel and 700 mg/m2 of cyclophosphamide. For patients with no prior chemotherapy, the MTDs were 75 mg/m2 of docetaxel and 800 mg/m2 of cyclophosphamide. The dose-limiting toxicity was neutropenic fever, observed in 41% of patients and 13% of cycles. Addition of granulocyte colony-stimulating factor (G-CSF, filgrastim [Neupogen]) did not permit further dose escalation, although it did result in briefer periods of neutropenia. PMID- 9213324 TI - Docetaxel/doxorubicin/cyclophosphamide in the treatment of metastatic breast cancer. AB - A pilot phase II study examined the feasibility of 75 mg/m2 of docetaxel (Taxotere) in combination with 50 mg/m2 of doxorubicin and 500 mg/m2 of cyclophosphamide (Cytoxan, Neosar) in the first-line treatment of metastatic breast cancer. This study was designed to evaluate the efficacy and toxicity of the docetaxel/doxorubicin/cyclophosphamide combination both alone and as induction before high-dose chemotherapy, supplemented by autologous peripheral blood stem-cell transplantation. Patients were divided into three groups: one group received 8 courses of docetaxel/doxorubicin/cyclophosphamide; the second received 4 to 6 courses of the same combination with cell sampling, followed by high-dose chemotherapy with autologous peripheral blood stem-cell transplantation; and the third group's regimen was identical to that of the second, with additional granulocyte-colony stimulating factor (G-CSF, filgrastim [Neupogen]). Of 28 patients (149 courses) evaluable for toxicity and response, the overall response rate was 82%, with 5 (18%) complete responses and 18 (64%) partial responses. The most frequent hematologic toxicity was neutropenia; grade 4 neutropenia occurred in 86% of patients, with febrile neutropenia in 9 patients (18%). There was no incidence of infection, possibly because of the administration of oral ciprofloxacin (Cipro) from days 5 to 15 of each cycle. Nonhematologic adverse events were not severe; there was no significant cardiotoxicity. Future randomized trials of docetaxel/doxorubicin/cyclophosphamide as first-line adjuvant therapy of high risk patients and as induction chemotherapy are in development. PMID- 9213325 TI - Docetaxel combined with vinorelbine: phase I results and new study designs. AB - This was a phase I dose-finding and pharmacokinetic study of vinorelbine (Navelbine) and docetaxel (Taxotere) as first-line chemotherapy for metastatic breast cancer. Vinorelbine dose, 20 or 22.5 mg/m2, on days 1 and 5, was followed on day 1 by docetaxel every 21 days, in doses increasing from 60 to 100 mg/m2. Two maximum tolerated doses were reached, the first at 75 mg/m2 of docetaxel and 22.5 mg/m2 of vinorelbine, and the second at 100 mg/m2 of docetaxel and 20 mg/m2 of vinorelbine. Symptomatic peripheral neuropathy was not observed. The recommended doses for phase II studies are 75 to 85 mg/m2 of docetaxel on day 1 and 20 mg/m2 of vinorelbine on days 1 and 5, every 3 weeks. The treatment regimen, which included 3-day corticosteroid prophylaxis, resulted in only mild fluid retention. Responses were seen at all dose levels, with an 80% overall response rate at the higher recommended dose; the overall response rate for patients at all dose levels was 66%. A high rate of response, including a complete response, was observed in patients with liver metastases. PMID- 9213327 TI - NSABP Protocol B-27. Preoperative doxorubicin plus cyclophosphamide followed by preoperative or postoperative docetaxel. AB - Protocol B-27, conducted by the National Surgical Adjuvant Breast and Bowel Project (NSABP), is a phase III, randomized trial designed to evaluate whether sequencing docetaxel (Taxotere) to neoadjuvant doxorubicin/cyclophosphamide (Cytoxan, Neosar) prolongs disease-free and overall survival in patients with operable breast cancer. Patients are being randomized into three groups. The control group receives four 21-day courses of doxorubicin/cyclophosphamide chemotherapy with tamoxifen (Nolvadex), followed by breast surgery (and postoperative radiation for patients receiving breast-conserving surgery). Two experimental groups receive the same doxorubicin/cyclophosphamide chemotherapy and tamoxifen, followed by docetaxel--either before (preoperative group) or after (postoperative group) surgery. In the first 11 months of the study, 283 patients- of a projected 1,606 patients over a 5-year period--have been entered. Slightly more than half of the patients are younger than 50 years of age. Nearly half of the patients presented with tumors that were more than 4.0 cm in greatest diameter. Biopsy was performed by fine-needle aspiration slightly more than half of the time. Slightly more than two-thirds of the patients had clinically negative nodes. Lumpectomy was the proposed surgery at entry in 40% to 43% of the patients. As of November 1996, toxicity information is available on 29 patients in the preoperative docetaxel group and 23 patients in the postoperative docetaxel group. So far, there have been no unexpected toxicities, but the data are too preliminary to report in detail. PMID- 9213326 TI - Docetaxel in combination with fluorouracil: study design and preliminary results. AB - The relatively recent introduction of a new class of chemotherapeutic agents--the taxoids--has raised hope of improved survival for patients with advanced or metastatic cancer. Following encouraging preclinical results of taxoid combinations, this phase I, nonrandomized trial was designed to evaluate a 1-hour intravenous infusion of docetaxel (Taxotere) on day 1 combined with fluorouracil (5-FU) as a daily intravenous bolus for 5 consecutive days. To date, 27 patients with advanced solid neoplasms have received 86 courses of docetaxel/5-FU at the following dose levels: 25/100, 35/150, 50/200, 60/200, and 60/300 mg/m2. Preliminary results showed no unexpected toxicities, and the principal toxicity was neutropenia of short duration. A treatment regimen of 60 mg/m2 docetaxel on day 1 and 300 mg/m2 of 5-FU given for 5 days, with a single course length of 28 days, is projected as the maximum tolerated dose. PMID- 9213329 TI - P-glycoprotein mediated multidrug resistance and its implications for pathology. AB - The discovery of P-glycoprotein has revealed a fundamental mechanism by which cancer cells evade chemotherapy and this principle has proven relevant to general cellular defence mechanisms in normal physiology. To date this knowledge has promised to improve current cancer chemotherapy through the manipulation of drug combinations according to the P-glycoprotein status of the tumor. Furthermore, the discovery of inhibitors of the protein may provide new therapeutic tools in the treatment of multidrug resistant neoplasia, provided the benefits are deemed greater than the potential detrimental side effects. When looking towards future therapies, however, we must also consider additional mechanisms which undoubtedly contribute to clinical drug resistance. Complete elucidation of this complex cellular defence network will hopefully translate into therapeutic opportunities to circumvent all mechanisms of multidrug resistance, thus positively impacting on patient survival. PMID- 9213328 TI - The pathology of fatal child abuse. AB - It is a sad indictment of human society that the abuse of children is such a prevalent and widespread problem. The acknowledgement that physical, emotional and sexual injury as well as intentional neglect can be inflicted upon the young by any person but especially by caregivers has been increasingly realised by the community. As a result, many professionals, especially in the medical sciences, are involved in the study and management of such cases with the ultimate goals of recognising children at risk, diagnosing those cases that have occurred, preventing initial or subsequent injury and bringing perpetrators to justice. The aim of this paper is to review recent published work on the pathology of abuse leading to death of the child. Particular reference is made to the patterns of observed physical damage as well as to the interpretation of those observations. Clearly many more children are abused than die directly as a result of that abuse, but pathologists are infrequently involved in the management of clinical abuse cases. Exceptions to this rule, of course, include assessment of biochemical changes in cases of Munchausen syndrome by proxy, diagnosis of infective lesions resulting from sexual assault as well as the interpretation of unexplained cutaneous lesions subsequently shown to be caused by physical assault (such as burns and bite marks). Cases of physical abuse are usually managed by pediatric specialists with assistance from radiologists, neurosurgeons and ophthalmologists, and it is important that effective communication be maintained by pathologists with these practitioners when investigating a case that has unfortunately culminated in death. PMID- 9213330 TI - New insights into the control of cell growth; the role of the AxI family. AB - AxI, Dtk and Mer are recently described receptors that constitute a new receptor tyrosine kinase subfamily. They bind the vitamin K-dependent protein growth arrest-specific gene 6 (Gas6) that is structurally related to the anticoagulation factor protein S. Studies suggest a role for these receptors in developmental processes. In the function of the hematopoietic and nervous systems and in tumorigenesis. PMID- 9213331 TI - The AgNoR count in ulcerative colitis with and without dysplasia. AB - The numbers of specific intranuclear AgNOR silver precipitates (AgNORs) and their relation to the Inflammatory Bowel Disease-Dysplasia Morphology Study Group's classification were studied in 134 colonscopic biopsy specimens of ulcerative colitis (UC) with and without dysplastic findings from 73 patients with active and inactive disease. In addition, 40 cases of colonic adenocarcinomas and 22 normal colonic mucosae were also assessed for comparison with the one-step silver staining AgNOR technique. Statistical comparison of the mean AgNOR number revealed a highly significant inter-class difference (P < 0.001) between UC without dysplasia and UC with dysplasia as well as between active and inactive UC. On the contrary, the AgNORs count did not separate colonic adenocarcinomas from UC with dysplasia either high or low grade nor these three classes from the group of UC with indefinite dysplasia. The quantitative evaluation of epithelial changes occurring in UC by the AgNOR method may help to establish the clinical management of colitis patients, in addition to qualitative morphological criteria. PMID- 9213332 TI - Metallothionein: a potential marker for differentiating benign and neoplastic gastrointestinal lymphoid infiltrates. AB - Metallothioneins (MT) are low molecular weight, metal-binding proteins. The induction of MT synthesis by cytokines, hormones and other cytotoxic agents indicates its role in cellular proliferation and differentiation as well as in cellular defense mechanisms. Previous studies have detected expression of MT in various human tumors by immunohistochemical staining. In certain cases the presence of MT in a tumor may be associated with its resistance against radiation and chemotherapeutic agents. Immunohistochemical staining of MT using a rabbit polyclonal anti-rat liver MT antibody was carried out in eight gastric, two small bowel and one large bowel lymphomas, and in ten control gastrointestinal (gastric, colonic and small bowel) biopsies or excised bowel segments with benign lymphoid infiltrates. The primary antibody against rat liver MT readily cross reacts with human MT. The neoplastic cells in nine of 11 malignant lymphomas showed weak to intense staining for MT, mostly in cytoplasm. In these cells a few nuclei (less than 5% of all tumor cells) were stained positively for MT. The benign lymphocytes in the gastrointestinal excised specimens and biopsies were mostly negative; four cases showed few positive cells. The giant cells were also positive for MT in a Crohn's disease case. We conclude that the presence or absence of MT in lymphocytes, detected by immunohistochemistry, may indicate the growth patterns of these cells. The distinct pattern of MT staining in malignant lymphoma in our study is suggestive of a potential use of MT staining as a discriminator between benign and malignant lymphoid tissues. PMID- 9213333 TI - p53 and proliferating cell nuclear antigen in endocrine tumors of pancreas and intestinal carcinoids. AB - Twenty six endocrine tumors of the pancreas and 19 carcinoids of the intestinal tract were studied with immunocytochemical staining for p53 and proliferating cell nuclear antigen (PCNA) to detect any correlation between PCNA and p53 immunostaining. PCNA immunostaining is somewhat variable in endocrine tumors of the pancreas whereas p53 overexpression was detected in one out of eight insulinomas (12.5%), four out of eight gastrinomas (50%) and none of two glucagonomas, six PPomas and two non-functioning tumors. In 19 intestinal carcinoids, two of eight small-intestinal carcinoids (25%) were positive for p53 whereas five appendiceal and two colonic carcinoids were negative for p53. It was concluded that: 1. PCNA staining is variable with no clear distinction between benign and malignant endocrine tumors of the pancreas and intestinal carcinoids, although positive PCNA staining may assess growth fraction of tumors; 2. p53 overexpression is relatively rare both in endocrine tumors of the pancreas (one out of 18 non-gastrinomas) and intestinal carcinoids, except for gastrinomas, in which 50% were positive for p53; and 3. non-neoplastic islet cells are positive for p53. PMID- 9213334 TI - p53 oncoprotein accumulation in adenoid cystic carcinoma of parotid and palatine salivary glands. AB - Previous studies have suggested that alterations of the p53 gene are the most common genetic abnormality in human cancer. The aims of the present study were to evaluate p53 protein (p53P) immunostaining in adenoid cystic carcinoma (ACC) of the salivary gland and to correlate the expression with patient survival. A total of 27 cases of ACC in the parotid gland (n = 12) and the minor palatine glands (n = 15) were studied, with ten cases each of normal parotid and palatine glands as non-neoplastic controls. Staining was performed with mouse monoclonal antibody DO 7 against p53 (Dako, USA) using the ABC method. Stained nuclei irrespective of intensity or frequency were considered as positive. The frequency of positive nuclei was evaluated as the p53P index (p53PI), the percentage of the total nuclei in the reference epithelium. Clinical survival data were available for patients for periods up to 156 months. Our data showed that no normal tissues showed immunoreactivity with p53P in their nuclei. Thirteen of 15 (87%) cases of palatal and two of 12 (17%) cases of parotid neoplasms stained with p53P and the p53PI ranged from 0.01 to 10%. The number of p53P positive tumors was significantly higher in palatal than in parotid neoplasms, suggesting that palatal ACCs may be more aggressive in comparison with parotid ACCs. Our data also showed that the number of p53P positive tumors was significantly increased in patients who died of tumors than in patients with no evidence of disease at the end of the follow-up period between 60 to 156 months. These results suggest that p53P may be involved in the development of salivary gland ACCs and that p53P analysis may be a useful indicator of poor prognosis. PMID- 9213335 TI - A clinicopathologic study of 30 cases of oligoastrocytoma including p53 immunohistochemistry. AB - Mixed gliomas have been difficult to define and subsequently diagnose due to the paucity of literature specifically examining this group of tumors. Thirty mixed gliomas in which the minor glial component comprised at least 20% of the total tumor were studied: 20 oligoastrocytomas (OA) and ten malignant oligoastrocytomas (MOA). Nineteen patients were male (mean age 36 years) and 19 patients (63%) presented with seizures. The tumor was located in the frontal lobe in 17 patients (57%) and the temporal lobe in nine patients (30%). The duration of preoperative symptoms in 25 patients ranged from five days to 14 years (mean 2.6 years). A mean follow-up of four years was available in 29 patients. Fourteen patients, seven with OA and seven with MOA, had recurrent tumor. One patient with MOA and four patients with OA (three with tumor progression and one with extensive leptomeningeal spread) died as a result of their tumor one to five years after diagnosis. Eighteen patients received chemotherapy and/or radiation therapy. Twenty-five tumors were immunostained with antibody to p53 protein. p53 nuclear staining was seen in 5/16 OA (31%) and 3/9 MOA (33%). Positive staining was observed only in astrocytic appearing cells. One of the four patients who died with OA was p53 positive. Three recurrent MOAs were p53 positive. Our study indicates that: 1. mixed gliomas most frequently occur in the frontal lobe and the majority of patients present with seizures; 2. there is no obvious association of p53 detection in mixed gliomas with tumor grade or behavior; and 3. similar to pure fibrillary astrocytomas, a subset of OA and MOA may be associated with p53 alterations. PMID- 9213336 TI - Broadsheet number 41: frozen section and intra-operative diagnosis. PMID- 9213337 TI - Test and teach. Number eighty three: Part 1. Idiopathic arterial calcification of infancy. PMID- 9213338 TI - The MDA-180 coagulation analyser: a laboratory evaluation. AB - We have evaluated the technical performance of the MDA-180 automated coagulation analyser in a working diagnostic hemostasis laboratory environment. The analyser has been on site now for over 18 months, and has undergone considerable testing. More than 22,000 samples have been processed, with over 90% of these via the MDA 180's cap-piercing facility. The instrument has been primarily assessed for its technical ease of use and continued reliability, as well as its analytical performance. The instrument has also been successfully interfaced to, and used with, our Laboratory information (CERNER PATHNET) system. A major feature of our evaluation has been an assessment of the MDA-180's ability to perform assays currently performed using alternative methodology or instrumentation (eg; ELISA methodology or the Coagamate-X2 and ACL-300R instruments), as well as its potential to streamline the technical performance of some of these assays. We have co-evaluated the following assays: PT/INR, APTT, TT, Fibrinogen, Protein C, ATIII, Factors II, V, VII, VIII, IX, X, XI, and XII, Lupus anticoagulant (dRVVT), and heparin (alpha Xa). In addition, a number of different reagents (particularly for PT and APTT assays) have been tested on the instrument. Intra-assay and inter assay variation appears to be remarkably low (five different plasmas tested: PT: 0.6 to 1.3% and 0.5 to 1.3% respectively; APTT: 0.7 to 3.2% and 0.6 to 3.6% respectively; single day analysis). Other comparative assessment data typically showed good correlation to existing test assay systems. A review of other features which may enhance or detract from the instrument's worth in a given hemostasis laboratory is also presented. In summary however, we conclude that the instrument is reliable, easy to use and capable of fast sample through-put. PMID- 9213339 TI - Autoantibody formation after allogeneic bone marrow transplantation: correlation with the reconstitution of CD5+ B cells and occurrence of graft-versus-host disease. AB - Manifestations of autoimmune diseases are common in patients who have received allogeneic bone marrow transplantation (BMT). Autoantibodies have been reported in these patients but the source and clinical significance of these autoantibodies are still obscure. In the present study the kinetics of autoantibody formation and the reconstitution of CD5+ B cells was followed in 21 patients who were submitted to allogeneic BMT. Anti-nuclear, anti-smooth muscle, anti-neutrophil cytoplasmic antibodies, anti-reticulin and rheumatoid factor were found at a frequency of 25%, 17%, 24%, 22% and 10% respectively after BMT. Anti double stranded DNA levels were mildly elevated in 15% of samples. The screening for anti-extractable nuclear antigen, anti-mitochondrial, anti-gastric parietal cell, anti-proteinase III, anti-myeloperoxidase, anti-lactoferrin antibodies was negative. The percentage and absolute count of CD5+ B cells increased with time after allogeneic BMT. Those patients with anti-nuclear or anti-smooth muscle antibodies had significantly higher CD5+ B cell counts than those without these two antibodies. Correlations of CD5+ B cell counts with other autoantibodies were negative. Acute graft-versus-host disease (GVHD) occurred in eight of the patients and chronic GVHD in four patients, but the frequency of autoantibodies had no relationship with the occurrence of acute or chronic GVHD. PMID- 9213340 TI - Characterization of platelet aggregation induced by the human carcinosarcoma Colo 526: role of platelet activation, tumor cell cytoskeleton and tumor cell plasma membrane. AB - Tumor cell-platelet interactions have been shown to be involved in the process of metastasis. This study characterizes the aggregation of washed platelets induced by the human uterine carcinosarcoma Colo 526. Ultrastructural studies revealed a two-stage process in which the earliest events were the adhesion and degranulation of individual platelets in contact with the tumor cell membrane. The second stage consisted of a wave of aggregation involving all residual platelets. We found that the first stage was initiated by a factor integral to the tumor cell plasma membrane which acted independently of the tumor cell cytoskeleton or metabolic processes. This factor was found to be a glycoprotein or glycolipid with functionally important sialic acid and N-linked carbohydrate residues. The initial stage was not dependent on platelet activation as neither aspirin nor prostacyclin prevented adhesion or degranulation. The second stage was found to be dependent on platelet activation. These results suggest that platelet aggregation induced by Colo 526 involves a distinctive primary stage which is initiated by a factor on the tumor cell plasma membrane resulting in the degranulation and lysis of individual platelets. This process can occur independently of platelet activation or aggregation and thus may have some relevance to the clinical use of platelet antagonists as antimetastatic agents. PMID- 9213341 TI - Costing pathology services: a practical approach to a difficult problem. AB - Recent developments in funding healthcare have created a need to know the cost of each input into a patient episode of care. In most cases pathology departments will be faced with a complex costing problem in their efforts to meet this need, owing to the diversity of investigations performed and the variation in cost between the least and the most expensive investigations. Most methods for costing pathology services are resource intensive, from the point of view both of the initial set up and of maintenance of the data. They often require a level of financial detail that may not be available. The model described in this paper will accept crude financial data and can be refined as better data become available. In our hands the model has yielded useful information and has raised the awareness of staff to the cost of the work they perform. Although the model has been applied to an analysis of pathology costs, the principles are transferable to other areas which have a wide range of "products" for which individual costs may be difficult to identify. PMID- 9213342 TI - Typing of Neisseria meningitidis by restriction analysis of the amplified porA gene. AB - We tested a typing system for 54 isolates of Neisseria meningitidis using polymerase chain reaction (PCR) amplification of the porA gene. The isolates were obtained between 1989 and 1994 from cases in Western Australia and Sydney. The PCR product was digested by five restriction endonucleases (AluI, HaeIII, HinfI, RsaI and HpaII) giving a restriction fragment length polymorphism (RFLP) pattern for each isolate. All of the isolates were able to be assigned an RFLP pattern, whereas 24 could be fully serotyped and serosubtyped. The method was rapid and simple to perform and results were easy to interpret. Two outbreaks of invasive meningococcal disease were included in the analysis, one involving an hyperendemic focus of disease and the other characteristic of a point outbreak. The typing system demonstrated the genetic relatedness of isolates from the point outbreak and the genetic diversity among the hyperendemic strains. We conclude that the method is discriminatory and is a useful supplement to serological typing for studying Australian outbreaks of invasive meningococcal disease. PMID- 9213343 TI - Selection of optimum laboratory tests for the identification of Moraxella catarrhalis. AB - We evaluated a variety of conventional and rapid tests and examined the erythromycin susceptibility of a collection of Moraxella catarrhalis and commensal neisseria strains in order to determine the optimum method for routine identification. One hundred and fifty three strains were tested by Gram stain, catalase, oxidase, carbohydrate degradation by two methods and the presence of esterases using indoxyl acetate, 4-methylumbelliferyl butyrate (MUB). Tween 80 and tributyrin as substrates. Erythromycin MICs and zone diameters around 1, 5 and 15 micrograms discs were determined by the NCCLS method for 151 of the strains. A combination of Gram stain, oxidase and either indoxyl acetate, spot MUB or tributyrin hydrolysis test proved to be reliable and potentially the most convenient for routine testing. MICs and zone diameters easily distinguished between the erythromycin-sensitive M. catarrhalis and the erythromycin-resistant commensal neisserias and would provide confirmation of identification if used for susceptibility testing. PMID- 9213344 TI - Disc susceptibility testing for thermomphilic campylobacters. AB - The minimum inhibitory concentrations (MICs) and zone diameters around NCCLS strength discs of 100 clinical isolates of thermophilic Campylobacter species, including 79 strains of Campylobacter jejuni subsp. jejuni, 19 of C. coli and two of C. lari, plus three type strains of these species, were determined for erythromycin, clindamycin, nalidixic acid, norfloxacin, ciprofloxacin, ampicillin, piperacillin, cephalothin, ceftriaxone, chloramphenicol, gentamicin and tetracycline. Using error-rate bounded analysis and adjustment of MIC breakpoints to fit natural populations, tentative interpretive zone diameter criteria were set for each of the antimicrobials. Application of these criteria showed that resistance to quinolones was not detected in species other than C. lari. Two strains of C. jejuni subsp. jejuni were susceptible to cephalothin. The type strain of C. lari was susceptible to erythromycin and resistant to clindamycin. Full resistance to erythromycin, chloramphenicol or gentamicin was not found in any strain, while nine strains were resistant to tetracycline. This disc method should provide a simple approach to resistance detection for surveillance or routine testing of invasive isolates. PMID- 9213345 TI - Intra-abdominal pulmonary sequestration: diagnostic difficulties. AB - This case report describes the unusual occurrence of congenital cystic adenomatoid malformation (CCAM) type 2 in an intra-abdominal pulmonary sequestration as a cause of diagnostic difficulties. The mass was discovered incidentally during a routine prenatal ultrasound in an infant with no other congenital malformation. Extralobar pulmonary sequestrations (EPS) located in the abdomen are rarely diagnosed prior to excision. The presence of CCAM type 2 in this situation might impose difficulties in histologic diagnosis of EPS, especially at the time of frozen section as happened in this case. To our knowledge this case is the sixth reported case of intra-abdominal EPS and the sixteenth case of EPS histologically displaying the feature of CCAM type 2. Pathologists should be aware that EPS in any location might display histologic features of CCAM type 2 even if the EPS is intra-abdominal. PMID- 9213347 TI - Phyllodes tumor with lobular carcinoma in situ and liposarcomatous stroma. AB - We describe a patient with a malignant phyllodes tumor with both lobular carcinoma in situ and liposarcomatous stromal differentiation. Although lobular carcinoma in situ and adipose differentiation have been reported as separate features rarely seen in the phyllodes tumor, we are unaware of any cases in which both of these features have been seen within the same tumor. The prognosis for this patient related not only to the malignant stromal component of the tumor but also to her increased risk of developing carcinoma in the remaining breast tissue. PMID- 9213346 TI - Acquired high titre factor VIII inhibitor with underlying polyarteritis nodosa. AB - We here present the case of a 70-year-old woman referred to our unit for investigation of bleeding. Investigations confirmed a high titre acquired Factor VIII inhibitor. In association there was relapse of systemic illness associated with anti-neutrophil cytoplasmic antibodies (atypical pattern) for which she had been treated five years previously. Immunosuppression was attempted, but it failed to have an impact both on the inhibitor titre and on the underlying disorder. The patient died from multi-organ failure and massive chest hemorrhage. Post-mortem showed necrotizing vasculitis of medium sized vessels at several sites, including the kidney, consistent with a diagnosis of polyarteritis nodosa. Although it is well recognised that Factor VIII inhibitors are found in conjunction with autoimmune disorders, this case is significant in that it is the first associated with histologically proven polyarteritis nodosa type vasculitis. The case illustrates the difficulties in the investigation and management of patients with acquired high titre Factor VIII inhibitors. PMID- 9213348 TI - Giant cell arteritis of the uterus: case report and review. AB - We here report a rare case of giant cell arteritis (GCA) of the myometrium found incidentally in a 68-year-old Caucasian woman presenting with uterovaginal prolapse and a known past history of temporal arteritis/polymyalgia rheumatica. Histology revealed a segmental arteritis of small, medium and some quite large myometrial arteries with extensive destruction of both internal and external elastic laminae. Multinucleate giant cells, lymphocytes and histiocytes were most prominent in the inflammatory infiltrate. The findings in this case are compared with previous reports. In a review of the literature it was found that almost one third of cases presented with generalised symptoms such as fever, anemia, fatigue and weight loss. The symptoms were not immediately recognised as temporal arteritis or polymyalgia rheumatica. On routine physical examination or radiological investigation, benign gynecological pathology such as a simple ovarian cyst or uterine leiomyoma were found. The subsequent unexpected discovery of GCA on histological examination was the critical event in alerting clinicians to the diagnosis of temporal arteritis/polymyalgia rheumatica. Without exception steroid therapy was successful in achieving relief of generalised symptoms. PMID- 9213349 TI - Corynebacterium pseudotuberculosis is a cause of human necrotising granulomatous lymphadenitis. AB - Corynebacterium pseudotuberculosis is a well recognised pathogen of farm animals, particularly sheep and goats. Human infection is a rare occurrence. This report describes suppurative lymphadenitis occurring in an adolescent boy who had contact with farm animals. The histological differential diagnosis of suppurative granulomatous lymphadenitis is discussed, and the importance of lymph node culture is stressed. PMID- 9213350 TI - Congenital factor VII deficiency presenting as delayed bleeding following dental extraction. A review of the role of factor VII in coagulation. AB - Recent updated models of the coagulation mechanism suggest that factor VII/thromboplastin complex is the main initiator or trigger of coagulation. Factor VII activation of factor IX is likely to be an important activation pathway. Inhibition of factor VII by tissue factor pathway inhibitor may have a regulatory role in the initiation of coagulation. Defining factor VII's interactions in coagulation physiology may lead to answers for some clinical problems in both hemostasis and thrombosis. We describe a 15-year-old Chinese boy with factor VII deficiency and a factor VII level of 0.08 U/ml. His symptoms were recurrent epistaxis and moderate delayed bleeding post-dental extraction. Such specific symptoms have been reported previously in a study of 40 European patients. It is one diagnosis to consider if a patients main symptom is significant post-dental bleeding. It is possible that there is requirement for higher levels of factor VII at these anatomical sites making them the common sites for symptoms in patients with moderate deficiency. Case reports of rare clotting factor deficiencies will illustrate what may be important in vitro functions of clotting proteins. Therefore reporting should be encouraged, especially during review and reconsideration of models of the coagulation mechanism. PMID- 9213351 TI - Vertebral osteomyelitis due to Pseudallescheria boydii. AB - A case of primary vertebral osteomyelitis due to an opportunistic fungus, Pseudallescheria boydii, in a child with acute myeloid leukemia, is reported. To our knowledge this is the first such case in a child, and only the second reported case in the international literature of primary spinal osteomyelitis due to this organism. We discuss the problems presented by this case both in terms of diagnosis and management. We also discuss the role of surgery and systemic antifungal chemotherapy in the treatment of deep infections due to Pseudallescheria boydii. PMID- 9213352 TI - Desmoplastic melanoma of the vulva. AB - The clinical and pathological features of a nonulcerated desmoplastic melanoma in the vulva of a 52-year-old woman are presented. Pleomorphic neoplastic spindle cells, fibroblasts and collagen formed a poorly demarcated 18 mm dermal mass. No adjacent intraepidermal component was seen. Immunoreactivity was demonstrated for S100 protein, vimentin, neuron-specific enolase and actin, but not for HMB-45, CAM 5.2, cytokeratin, epithelial membrane antigen, desmin or CD34. Electron microscopic examination was noncontributory. Treatment included a left hemivulvectomy with ipsilateral groin node dissection followed by radiotherapy. The tumor recurred six weeks later and was unresectable. The patient is alive with symptoms nine months after presentation. This is the first case report of a vulvar desmoplastic melanoma without neural involvement or an intraepidermal component. The variable tumor cytomorphology, nonspecific immunohistochemical and ultrastructural features render a diagnosis more difficult than with other primary cutaneous melanomas. PMID- 9213353 TI - Cloning and characterization of the 5' flanking sequences (promoter region) of the human GLP-1 receptor gene. AB - The glucagon-like peptide 1 (7-36) amide (GLP-1) receptor mediates the insulinotropic effects of the incretin hormone GLP-1. To elucidate the tissue specific regulation of the GLP-1 receptor we screened a human genomic library with a human GLP-1 receptor cDNA. The gene spans 40 kb and consists of at least seven exons. The promoter contained no TATA- or CAAT-boxes, but several other putative cis-regulatory recognition sequences including three Sp1 binding sites. Transient transfections of GLP-1 receptor producing and non-producing cells with promoter/ reporter gene constructs revealed that the putative Sp1 binding sites and several other silencer and tissue specific elements are important for the activity. Therefore, 3000 bp upstream the GLP-1 receptor coding sequences comprise regulatory elements essential for the tissue- and cell-specific transcription of the gene. PMID- 9213354 TI - Endoproteolysis of glucagon-like peptide (GLP)-1 (7-36) amide by ectopeptidases in RINm5F cells. AB - This study concerns whether the pancreatic beta cell expresses cell-surface ectopeptidases that are capable of proteolysis of peptide hormones and neuropeptides that modify glucose-dependent insulin release. These biochemical investigations of the RINm5F cell line found that these cells express ectopeptidases. We have characterized the limited endoproteolysis of GLP-1 (7-36) amide that occurs in the presence of RINm5F plasma membranes. The products and the sensitivity to specific peptidase inhibitors of the proteolysis is characteristic of neutral endopeptidase (NEP) 24.11. Vasoactive intestinal polypeptide (VIP), pituitary adenylate cyclase-activating peptide (PACAP), amylin, glucagon, glucose-dependent insulinotropic polypeptide (GIP), and exendin 4 also undergo proteolysis in the presence of RIN cell membranes. NEP 24.11 activity in RIN cell membranes was confirmed using a specific fluorogenic assay, by histochemistry, and by comparison with the recombinant enzyme with respect to the kinetics of proteolysis of GLP-1 (7-36) amide and of a fluorogenic substrate. Specific fluorogenic assays revealed the presence of aminopeptidase N and the absence of aminopeptidase A and of dipeptidylpeptidase IV. PMID- 9213355 TI - Pure glucagon antagonists: biological activities and cAMP accumulation using phosphodiesterase inhibitors. AB - Five new glucagon analogues have been designed, synthesized, characterized and their biological activities tested. The investigation was centered on modifications in the N-terminal region in particular, residues at Thr5, Phe6 and Tyr10 positions, with the goal of obtaining pure glucagon antagonists in our newly developed high sensitivity cAMP accumulation assay. The structures of the designed compounds are: [des-His1, des-Phe6, Glu9] glucagon-NH2 (1); [des-His1, des-Phe6, Glu9, Phe10]glucagon-NH2 (2); [des-His1, Tyr5, des-Phe6, Glu9]glucagon NH2 (3); [des-His1, Phe5, des-Phe6, Glu9]glucagon-NH2 (4) and [des-His1, des Phe6, Glu9, D-Arg18]glucagon-NH2 (5). The binding potencies IC50 values in (nM) were 48.0, 27.4, 26.0, 20.0 and 416.0, respectively. All of these analogues when tested in the classical adenylate cyclase assay demonstrate antagonist properties, and in competition experiments, all caused a rightward-shift of the glucagon stimulated adenylate cyclase dose-response curve. The pA2 values for these analogues were 8.20 (1); 6.25 (2); 6.10 (3); 6.25 (4); and 6.08 (5), respectively. A newly revised assay has been developed to determine the intracellular cAMP accumulation levels in hepatocytes at the highest possible sensitivity. Four of the five glucagon analogues in this report (analogues 1, 2, 4 and 5), did not activate the adenylate cyclase in the presence of Rolipram up to a maximal physiological concentration of 1 microM, and thus are pure antagonists. PMID- 9213356 TI - Different influence of CGRP (8-37), an amylin and CGRP antagonist, on the anorectic effects of cholecystokinin and bombesin in diabetic and normal rats. AB - Because previous studies had suggested that the anorectic effects of cholecystokinin (CCK) and bombesin (BBS) depend partly on the release of amylin or calcitonin gene-related peptide (CGRP), we investigated the influence of the amylin and CGRP receptor antagonist CGRP (8-37) on the anorectic effects of CCK and BBS in streptozotocin (STZ)-diabetic and nondiabetic rats. STZ-diabetic rats had significantly lower plasma amylin and insulin concentrations than nondiabetic control rats. Amylin (5 micrograms/kg or 2.5 micrograms/rat) injected IP at dark onset after 24-h food deprivation elicited an anorectic effect of similar extent in STZ-diabetic and control rats. Under similar conditions, CCK (0.25 and 2 micrograms/kg) and BBS (5 micrograms/kg) reduced food intake in both STZ-diabetic and nondiabetic rats. These effects were markedly attenuated by CGRP (8-37) (10 micrograms/kg) in non-diabetics but not in STZ-diabetic rats. It is concluded that part of the anorectic effects of CCK and BBS depend on the release of amylin from pancreatic B-cells. PMID- 9213357 TI - Inhibition of insulin release by intraduodenally infused enterostatin-VPDPR in rats. AB - Enterostatin, an amino-terminal pentapeptide produced in the intestinal lumen after cleavage of pancreatic procolipase, has been shown to suppress fat intake in rats after intraduodenal infusion. In this study, female Sprague-Dawley rats fitted with a duodenal catheter were intestinally infused with enterostatin (Val Pro-Asp-Pro-Arg, 11.3 and 22.6 nmol/kg/min) plus 20% Intralipid for 30 min. Plasma insulin levels were significantly reduced, whereas plasma glucose concentrations were not altered by enterostatin-VPDPR. The tripeptide Asp-Pro-Arg was also found to decrease the levels of plasma insulin. However, the pentapeptide with the sequence Val-Pro-Gly-Pro-Arg, des-Arg-enterostatin Val-Pro Asp-Pro and the tripeptide Pro-Asp-Pro failed to cause the reduction of plasma insulin levels in rats following intestinal infusion of these peptides. Radiolabeled enterostatin ([3H]VPDPR) was identified in plasma by HPLC following intraduodenal infusion of the peptide, indicating that the appearance of an intact enterostatin-VPDPR in blood. It is concluded that intestinally administered enterostatin-VPDPR and its metabolites reduce plasma levels of insulin stimulated by Intralipid. PMID- 9213358 TI - Chronic ICV enterostatin preferentially reduced fat intake and lowered body weight. AB - The pancreatic peptide enterostatin will acutely reduce fat intake in rats provided a choice of diets. Chronic ICV infusions of enterostatin suppress the intake of high fat diet. However, the effects of chronic ICV enterostatin on diet choice has not previously been studied. To investigate this, enterostatin (0.5 microgram/h) or artificial cerebrospinal fluid (CSF) was infused for 9 days into the lateral ventricle of rats adapted to a two-choice high-fat (HF) and low-fat (LF) diet regime. Enterostatin reduced intake of HF diet with the maximum depression at day 4, but there was no compensatory increase in LF intake. The body weight of enterostatin-infused rats declined. This was associated with a reduction in fat pad and liver weights compared to the CSF-infused control rats. Serum triglycerides and insulin were decreased and corticosterone was elevated in enterostatin-infused rats. The data show that enterostatin will chronically reduce fat intake and body weight and suggest that enterostatin may attenuate the appetite for fat. PMID- 9213359 TI - Location and characterization of the human GRP receptor expressed by gastrointestinal epithelial cells. AB - The exact location of normal gastrin-releasing peptide (GRP) receptor expression by epithelial cells lining the human gastrointestinal (GI) tract is not known; yet this receptor is found on upwards of 50% of GI cancers. Furthermore, the pharmacology reported for GRP receptors expressed by GI cancers varies considerably. Therefore, the purpose of this study was to determine the normal distribution of GRP receptor expression by cells lining the human GI tract, and then determine the normal pharmacology of the human receptor when ectopically expressed by the nonmalignant human colon epithelial cell line NCM460. We obtained endoscopic pinch biopsies of, and extracted the RNA from, epithelial cells lining the esophagus, stomach, jejunum, ileum, and proximal and descending colon, RT-PCR demonstrated that GRP-R expression is limited to cells lining the gastric antrum, indicating that this receptor is aberrantly expressed by GI cancers. To determine the normal pharmacology of this receptor when expressed by nonmalignant human tissues for the first time, we transfected NCM460 cells with the cDNA for the human GRP receptor. By studying three stable NCM460 cell lines expressing varying numbers of receptors, we demonstrate that agonist and antagonist binding affinity, binding kinetics, and G-protein coupling are all independent of receptor number. Finally, by comparing GRP receptors expressed by GI cancers with those on NCM460-transfected cells, we show that the pharmacology of the aberrantly expressed receptors is significantly altered. Thus, these data demonstrate that GI cancers aberrantly express GRP receptors that then behave abnormally. PMID- 9213360 TI - Muscarinic control of phase III contractions and motilin release in dogs. AB - The role of muscarinic receptor subtype(s) in the control of gastric phase III contractions and motilin release was determined in dogs. Pirenzepine (226 nmol/kg/h) shortened the phase III cycle, causing an early increase in plasma motilin concentrations. Both AF-DX116BS and 4-DAMP inhibited the occurrence of spontaneous and motilin-induced phase III contractions, and 4-DAMP also inhibited the spontaneous increase in plasma motilin concentration. Only 4-DAMP inhibited carbachol-induced motilin release in perifused duodenal mucosal cells. In conclusion, M1 receptors are involved in the indirect inhibition of motilin release. Motilin-induced contractions are mediated mainly by M3 receptors and partly by M2 receptors, and M3 receptors are present in motilin-producing cells. PMID- 9213361 TI - Transepithelial transport of oligopeptides in the human intestinal cell, Caco-2. AB - The transepithelial transport of oligopeptides (of more than 4 residues) was studied by using human intestinal Caco-2 cell monolyers. The susceptibility to the brush-border peptidases was observed to be one of the primary factors which decide the transport rate. The apical-to-basolateral transport mechanism was investigated by using bradykinin and GGYR which were resistant to cellular peptidases. The intracellular pathway, probably the adsorptive transcytosis, was suggested to be involved in the transport of bradykinin and its analogues, the transport rate being particularly dependent on the hydrophobic properties of the peptides. On the other hand, the tetrapeptide such as GGYR was suggested to be transported mainly via the paracellular pathway. PMID- 9213362 TI - Cyclo (His-Pro) modulation of body temperature at hot ambient temperature in the desert rat (Mastomys natalensis). AB - Cyclo(His-Pro) (CHP) has been shown to facilitate cold-induced hypothermia in the desert rat Mastomys natalensis. In the present study, we examined the role of endogenous CHP in hyperthermia induced by hot ambient temperature (40 degrees C) in the above rodent species. The results of these studies show that housing rodents at 40 degrees C resulted in a altered distribution of CHP in the brain, with a rise in hypothalamic content accompanied by an increase in rectal temperature. While administration of exogenous CHP decreased hyperthermia, immunoneutralization of endogenous CHP increased hyperthermia. The results of these studies show that changes in endogenous CHP levels may affect body temperature regulation. PMID- 9213363 TI - Effects of some 7-arylidene and 7-heteroarylidene morphinan-6-ones on the antinociceptive activity of [D-Pen2, D-Pen5]enkephalin and [D-Ala2, Glu4]deltorphin II and on multiple opioid receptors. AB - The in vivo and functional effects of several 7-arylidene and 7-heteroarylidene morphinan-6-ones were determined at the mu-, delta-, and kappa-opioid receptors using the guinea pig brain membranes, guinea pig ileum (GPI), and mouse vas deferens (MVD). In vivo effects included the antagonism by these compounds given subcutaneously on the antinociceptive actions of intracerebroventricularly injected [D-Pen2, D-Pen5]enkephalin (DPDPE) and [D-Ala2, Glu4]deltorphin II (deltorphin II), the highly selective putative delta 1- and delta 2-opioid receptor agonists. Finally, the partition coefficients of these compounds were estimated (CLOGP) and determined experimentally at pH 7.4 in the 1-octanol/water system. Compared with E-7-benzylidenenaltrexone (BNTX), most compounds except for E-7-(4-chlorobenzylidene)naltrexone, were more potent at delta-opioid receptors than at the mu-opioid receptor, whereas, in comparison to the kappa-opioid receptor, the activities of the E-7-arylidene or E-7-heteroarylidene naltrexone derivatives at the delta-receptor were in the following order, where the 7 substituents were: 4-fluorobenzylidene- > benzylidene > 3-pyridylmethylene- > 4 pyridylmethylene- > 1-methyl-2-imidazolylmethylene > 4-chlorobenzylidene. In the MVD preparation, the potencies at the delta-opioid receptor, in comparison to BNTX, were in the following order, where the 7-substituents were: benzylidene = 1 methyl-2-imidazolylmethylene- > 4-fluorobenzylidene- = 3-pyridylmethylene- = 4 pyridylmethylene-. All compounds antagonized delta 1, and delta 2-opioid receptor agonist-induced analgesia. The antagonist potencies at the delta 1-opioid receptor were in the following order, where the 7-substituents were: benzylidene- > 4-chlorobenzylidene- > 4-fluorobenzylidene- > 3-pyridylmethylene- > 1-methyl-2 imidazolymethylene- approximately 4-pyridylmethylene-, whereas at the delta 2 opioid receptor, the order was benzylidene- > 4-chlorobenzylidene- > 4 fluorobenzylidene- > 3-pyridylmethylene- > 1-methyl-2-imidazolymethylene- > 4 pyridylmethylene. In general, all compounds exhibited greater potency at the delta 2- than delta 1-opioid receptor. The computed partition coefficients were, as expected, greater than the apparent log P values, which were determined experimentally. Generally, the lipophilicity values in decreasing order were: 4 chlorobenzylidene- > 4-fluorobenzylidene- > benzylidene > 3-pyridylmethylene- = 4 pyridylmethylene- > 1-methyl-2-imidazolylmethylene-. In general, the benzylidene and 4-pyridylmethylene derivatives, which have medium lipophilicities, were equally effective at the delta 1- and delta 2-receptors; the 3-pyridylmethylene and 1-methyl-2-imidazolylmethylene derivatives had lower lipophilicities and were more selective for the delta 2- than delta 1-receptor; the 4-chlorobenzylidene and 4-fluorobenzylidene derivatives were more lipophilic and had intermediate activity. The plot of pED50 values for the in vivo tests for the delta 1- and delta 2-receptors showed that the two receptors are not independent with respect to this series of compounds. PMID- 9213364 TI - Pulsed magnetic field induced "analgesia" in the land snail, Cepaea nemoralis, and the effects of mu, delta, and kappa opioid receptor agonists/antagonists. AB - A brief exposure to a pulsed magnetic field (Cnp: patent pending) had significant antinociceptive or "analgesic" effects in the land snail, Cepaea nemoralis, as evidenced by an increase in the latency of response to a warmed (40 degrees C) surface. This analgesia was in part opioid mediated being significantly reduced, but not eliminated: by the prototypic opiate antagonist, naloxone; the mu (mu) opioid receptor directed antagonists, naloxazine or beta-funaltrexamine, and the delta (delta) opioid receptor directed antagonists, naltrindole-5'-isothiocyanate or ICI 174,864. However the Cnp induced analgesia was unaffected by the kappa (kappa) opioid receptor directed antagonist, nor-binaltorphimine. The delta 1 and delta 2 opioid receptor directed agonists, (DPDPE, [D-Pen2,D-Pen5]enkephalin), (deltorphin, [D-Ala2,Glu4]), respectively, also had significant differential analgesic effects, supporting a functional delta opioid receptor mediated enkephalinergic mechanism in Cepaea. These results suggest that this specific pulsed magnetic field (Cnp) elicits significant analgesic effects through mechanisms that, in part, involve delta and, to a lesser extent mu opioid receptors. PMID- 9213365 TI - Enhancement of performance in multiple learning tasks by corticotropin-releasing factor-binding protein ligand inhibitors. AB - Evidence favors a role for corticotropin-releasing factor (CRF) in learning and memory processes. A binding protein (CRF-BP) with the ability to inactivate CRF provides a novel target to modulate endogenous levels of CRF. The present studies employed three measures of information processing in rats in order to examine the impact of CRF system activation resulting from administration of CRF-BP ligand inhibitors, which increase levels of "free CRF." Acquisition of a visual discrimination paradigm and retention of a inhibitory avoidance task were dose dependently facilitated by central administration of a CRF-BP ligand inhibitor. CRF-BP ligand inhibitor treatment also improved performance in an active avoidance paradigm in aged animals. No nonspecific anorexic effects of the active dose of CRF-BP ligand inhibitor were detected in a food intake test. Moreover, the magnitude of in vivo efficacy of the CRF-BP ligand inhibitor peptide in producing a mild increase in motor activity was dissociated from that of a postsynaptic CRF receptor agonist that exerted robust and long-lasting activity increases. Thus, CRF-BP ligand inhibitors appear to elicit generalized learning enhancement effects without mimicking the robust nonspecific behavioral actions of a CRF receptor agonist. PMID- 9213366 TI - Co-release of substance P and neurokinin A from the Atlantic cod stomach. AB - The function of tachykinins in the control of gastric motility in the cod, Gadus morhua, was studied using native cod substance P ([Lys1, Arg3, Ile3]SP) and cod neurokinin A ([Ile3, Asn4]NKA). Both cod SP and NKA produced contractions of the vascularly perfused cod stomach, SP being almost 6 times more potent than NKA (pD2-values 7.05 +/- 0.06 and 6.28 +/- 0.09, respectively). The release of tachykinins from the cod stomach was measured in radioimmunoassay, using specific antibodies for the two cod tachykinins. Stimulation of the stomach motility by electrical stimulation of the vagus nerve or infusion of acetylcholine increased the amounts of SP and NKA released into the vascular perfusate. The results suggest that both tachykinins are involved in the excitatory response of the cod stomach produced by vagal and cholinergic stimulation. PMID- 9213367 TI - Isolation and structural characterization of pituitary adenylate cyclase activating polypeptide (PACAP)-like peptide from the brain of a teleost, stargazer, Uranoscopus japonicus. AB - Pituitary adenylate cyclase-activating polypeptide (PACAP) is a novel neuropeptide consisting of 38-residue (PACAP 1-38) and a truncated form with 27 residues (PACAP 1-27) that plays several roles in tetrapods. We isolated a highly purified PACAP-like peptide from the brain of a teleost, the stargazer, by extracting of acetone-dried powder with acetic acid followed by high-performance liquid chromatography (HPLC) on gel-filtration, cation-exchange, and reverse phase columns. Purification was monitored by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting analysis using an anti-PACAP 1-27 antiserum. The PACAP-like peptide thus obtained had a molecular mass of 4,623, determined by mass spectrometry, and its amino acid sequence showed 89 and 87% identity with those of ovine and frog PACAPs, respectively. These results indicate that a PACAP-like peptide, which is a highly homologous with tetrapod PACAP, is present in the teleost brain. PMID- 9213368 TI - Nociceptin, an endogenous ligand for the ORL1 receptor, decreases cardiac output and total peripheral resistance in the rat. AB - The heptadecapeptide nociceptin, also known as Orphanin FQ, is a newly discovered endogenous ligand for the opioid-like G-protein coupled receptor, ORL1. In the present study, responses to intravenous injections of nociceptin were investigated in the systemic vascular bed of the rat. Nociceptin induced dose related decreases in systemic arterial pressure and total peripheral resistance when injected in doses of 1-30 nmol/kg i.v.. Nociceptin decreased heart rate and in doses of 10 and 30 nmol/kg i.v., significantly decreased cardiac output. In terms of relative vasodilator activity, nociceptin was approximately 10-fold less potent than the beta-adrenergic receptor agonist isoproterenol. These data show that nociceptin has novel vasodilator activity in the systemic vascular bed of the rat. PMID- 9213370 TI - Interindividual variability of enkephalin-degrading enzymes in human plasma. AB - The interindividual variability of the hydrolysis of leucine enkephalin, and of the formation of its hydrolysis by-products has been studied in human plasma. In agreement with known data, the data obtained indicate that Leu-enkephalin is degraded by several enzymes, belonging to three classes: aminopeptidases, dipeptidylaminopeptidases, and dipeptidylcarboxypeptidases. The relative ratio of the substrate degraded by each enzyme class-as well as the expression of the single enzyme species within each class-appears to be individually determined. Interindividual variability observed seems controlled by two main factors: the pattern of enkephalin-degrading enzymes and, more notably, the low molecular weight plasma inhibitors. Both these factors appear to be partially specific of each donor. Possibly because of the composition of these factors, the hydrolysis pattern of the substrate is characteristic of each donor, and constant in blood obtained from successive drawings, at least within a relatively short period of time. PMID- 9213371 TI - Peptide detection in single cells using a dot immunoblot assay. AB - A highly sensitive dot immunoblot assay (DIA) for the detection and quantitative measurement of small peptides in single cells is presented. This DIA protocol is simple, rapid, and produces no radioactive waste. Its femtomole sensitivity is 100 fold greater than previously described DIAs. This DIA method is sufficiently sensitive to allow reliable peptide measurements to be obtained from a single cell in a manner than is faster and easier than other peptide detection procedures. This method can also be used for several other purposes, including assessing antibody specificity and peptide quantification. PMID- 9213369 TI - Degradation of human adrenomedullin(1-52) by plasma membrane enzymes and identification of metabolites. AB - Very little is known about degradation or metabolism of adrenomedullin. To this end, we incubated adrenomedullin with ovine adrenal, kidney and lung plasma membrane preparations and showed the major degradation products were ADM(2-52) and ADM(8-52). Smaller amounts of ADM(26-52), (27-52), (28-52) and (33-52) were also produced. Degradation was inhibited by EDTA and 1,10 phenanthroline but not by phosphoramidon, leupeptin and pepstatin. The above data are consistent with initial hydrolysis adjacent to hydrophobic residues by a metalloprotease, generating ADM(8-52), (26-52) and (33-52), followed by an aminopeptidase action to produce ADM(2-52), (27-52) and (28-52). Improved understanding of the metabolism of ADM may have therapeutic implications, for example in the treatment of heart failure. PMID- 9213372 TI - Immunohistochemical localization of the pro-peptide processing enzymes PC1/PC3 and PC2 in the human anal canal. AB - The distribution of prohormone/pro-peptide convertases PC1/PC3 and PC2 was investigated in the human anal canal by immunohistochemistry. Both prohormone convertases exhibited region-specific distribution patterns and were observed in neural and neuroendocrine cells and in nonneuroendocrine cellular elements. PC1/PC3 immunoreactivity was present in enteric neurons, subsets of nerve fibers, and neuroendocrine cells, and also in epithelial cells like intestinal stem cells, and a subpopulation of squamous cells. Enteric neurons were PC2 immunoreactive, whereas PC2 immunostaining in nerve fibers was slightly above background levels. Few neuroendocrine cells contained PC2 immunoreactivity, which were located predominantly in the anal transitional zone. In the squamous epithelium, the basal cell layer stained for PC2. The tissue-specific distribution of PC1/PC3 and PC2 indicates region-specific processing of peptides with regulatory functions in the anal canal and further supports the hypothesis that neuropeptides are important regulators of anal functions. PMID- 9213373 TI - Selectivity of [Phe-I7], [Ala6], and [D-Ala4,Gln5,Tyr6] substituted ACTH(4-10) analogues for the melanocortin receptors. AB - We tested [Ala6]ACTH(4-10) and [Phe-I7]ACTH(4-10)(putative MC receptor antagonists), [D-Ala4,Gln5,Tyr6]ACTH(4-10)(BIM 22015), and ACTH (4-10) with radioligand binding using transiently expressed human MC1, MC3, MC4, and MC3 receptors. [Phe-I7]ACTH(4-10) had higher affinity for the MC3, MC4, and MC3 receptors but lower for the MC1 compared to ACTH(4-10). [Ala6]ACTH(4-10) did not bind the MC1 receptor but had highest affinity for the MC4 receptor. The data indicate that the His6 has a specially important role in binding to the MC1 receptor. The BIM 22015 did not bind to these MC receptor subtypes, which indicates that the neurotrophic and myotrophic properties that are attributed to this peptide are mediated by some other receptor. PMID- 9213374 TI - The changing prognosis of ARDS. AB - It seems likely that the prognosis of ARDS has indeed improved. To what extent is still uncertain, and unfortunately, from what is unknown. As a result, we don't know where to concentrate additional effort and resources. If survival has improved, changes in supportive therapy are the most likely cause. Thus, it is cynical and misdirected to assume that all methods of "supportive" care are equivalent, and that efforts to optimize such care are less worthy than those directed toward potentially specific causes. The situation is analogous to that seen previously in the management of myocardial infarction. Although breakthroughs in thrombolysis and angioplasty have occupied our attention in the past decade, the mortality from myocardial infarction fell dramatically in the decades that preceded these new therapeutic advances. All such improvements are to be hailed and appreciated, and further improvements, both in supportive and specific therapy should be anticipated as likely and welcome. PMID- 9213375 TI - The eosinophilia myalgia syndrome: to be or not to be. PMID- 9213376 TI - Is methotrexate oncogenic in patients with rheumatoid arthritis? PMID- 9213377 TI - Eosinophilia myalgia syndrome. AB - The term eosinophilia myalgia syndrome (EMS) was coined in 1989 after a cluster of cases with symptoms of incapacitating myalgias and eosinophilia were reported. This syndrome has been only defined as varying degrees of myalgias and peripheral eosinophilia. Case reports of EMS are protean and do not show a consistent clinical picture, raising the question of whether this reflects a single disorder or is a conflation of various disorders. There have been only two studies evaluating the association of EMS with 1-tryptophan. These two included only 23 patients with EMS. Apart from the obvious statistical fragility inherent in such small studies, each is further weakened by differences in the mechanisms by which patients and controls were selected. Furthermore, the continued reports of EMS after 1-tryptophan was removed from the market raise additional questions about the association. Nonetheless, there has been an inordinate reliance on a history of 1-tryptophan ingestion in making the diagnosis of EMS. When presented case studies, clinicians were much more likely to make the diagnosis of EMS when a history of 1-tryptophan ingestion was included. These observations underscore the need for careful application of well-considered diagnostic criteria to the study of new syndromes and their potential associations. PMID- 9213378 TI - Lymphoma in patients with rheumatoid arthritis: association with the disease state or methotrexate treatment. AB - Although long-term clinical studies have shown no excessive risk of lymphoma in rheumatoid arthritis (RA) patients treated with methotrexate (MTX), an increasing number of reports of this association continue to appear. We describe two cases, review the cases in the world's literature, and summarize their important characteristics. Possible oncogenic mechanisms are discussed. Most lymphoproliferation cases presented here have features of immunosuppression associated lymphoma. The immunosuppressed state is attributable to a combination of factors, such as RA itself and the actions of MTX. The risk factors for RA patients to develop lymphoma while on MTX include severe disease, intense immunosuppression, genetic predisposition, and an increased frequency of latent infection with prooncogenic viruses such as Epstein-Barr virus (EBV). The spontaneous remission of lymphomas in eight RA patients after MTX was stopped highlights the likely causative role of the drug in the development of these malignancies. If the clinical situation permits, a period of observation for spontaneous remission after MTX is stopped is advisable. The physicians caring for RA patients on MTX should maintain a high surveillance for signs and symptoms suggestive of lymphoma. PMID- 9213379 TI - Group education for rheumatoid arthritis patients. AB - This article reviews the effectiveness of group education programs in improving the knowledge, behavior, and health status of patients with rheumatoid arthritis (RA) and evaluates to what extent various programs fulfill certain criteria for educational self-management programs. Thirty-one studies are reviewed: in 12, patients with various rheumatic diseases including RA were included, and in 19, only RA patients were studied. Group education increased the knowledge of the participants, which was maintained over long intervals. Beneficial behavioral effects were found in mixed populations but less often found in RA patients. Group education often improved physical health status both in mixed and in RA populations, but seldom led to improved psychosocial health status. In general, the beneficial effects of group education were found more often in mixed populations than in strictly RA patients. Further investigations must examine which mechanisms make educational interventions effective and determine the types of interventions or combinations of interventions that are effective. Effects of group education on health status are almost never maintained over long intervals. More research is needed to develop strategies for maintaining and enhancing early gains from group education. PMID- 9213380 TI - Oral contraceptives and rheumatoid arthritis: results from a primary care-based incident case-control study. AB - OBJECTIVE: The possibility that oral contraceptives offer a protective effect against the development of rheumatoid arthritis is still contentious. Of the 17 studies investigating this association, 11 have found a protective effect, and 6 have not. These differences are probably attributable to either selection or information biases in a subset of studies, although the exact reason is unknown. To overcome the methodological problems inherent in the design of previous studies, we have conducted a population-based case-control study. METHODS: Women who were incident cases of inflammatory polyarthritis, defined as swelling of at least two joint areas lasting at least 4 weeks, were recruited directly from primary care and compared with age-matched women from the same population. RESULTS: Cases and controls reported a similar level of "ever use" of oral contraceptives, adjusted odds ratio = 0.88 (95% confidence interval, 0.47, 1.64). The cases were, however, less likely to report using oral contraceptives at the time of onset, adjusted odds ratio = 0.22 (95% confidence interval, 0.06, 0.85). Similar results were observed for cases who satisfied the criteria for rheumatoid arthritis and cases who did not. CONCLUSION: These results indicate that only current oral contraceptive use protects against the development of inflammatory polyarthritis. PMID- 9213381 TI - A role for histamine receptors in rheumatoid arthritis. AB - Although neurotransmitters and various chemical mediators play an important role in the pathogenesis of rheumatoid arthritis (RA), precise underlying mechanisms have yet to be determined. Histamine is a classical mediator of inflammation and three types of receptors are known. We investigated the presence and functions of histamine receptors of lymphocytes, bone marrow cells, synovial fibroblasts, and chondrocytes in experimentally-induced arthritis and human RA. The function of H2 receptors in peripheral blood lymphocytes and bone marrow cells were down regulated as measured by increments of intracellular cAMP and IL-6 production. Synovial fibroblasts from RA patients did not respond to H2 agonist to synthesize hyaluronic acid. It is evident that H2 receptors are down-regulated in lymphocytes, bone marrow cells, and synovial fibroblasts. The reduced function of H2 receptors in collagen-induced arthritis was normalized by transfer of the receptor-bearing lymphocytes and bone marrow cells. These data suggest that histamine is involved in the pathogenesis of RA. PMID- 9213382 TI - Quality of life and physical functioning of relatives of fibromyalgia patients. AB - OBJECTIVES: The quality of life (QOL) and health status of fibromyalgia syndrome (FS) patients is impaired, and may adversely affect their close relatives. The aim of this study was to assess the QOL and physical functioning of relatives of FS patients. METHODS: A total of 118 relatives (parents, husbands, siblings, and offspring) of 30 FS female patients were evaluated using a QOL scale and the Fibromyalgia Impact Questionnaire (FIQ) and were compared with 124 healthy controls. These measures of functioning and QOL were further studied in relatives with and without FS. RESULTS: Although the QOL of the relatives was better than that of the FS index women, they were significantly less satisfied than the controls with functioning-related aspects, namely work (job or home), independence, and health (P < .05). Relatives with FS (n = 29) and female relatives (n = 40) reported lower QOL than relatives without FS (n = 89) and male relatives (n = 78), respectively. Similarly, physical functioning of relatives, though better than in FS index cases, was significantly worse than in healthy controls. Furthermore, the health status of female relatives and relatives with FS was significantly worse than that of male relatives and relatives without FS, respectively. CONCLUSIONS: The quality of life and physical functioning of relatives of FS patients were found to be impaired, especially in female relatives and those with undiagnosed FS. This finding may be attributed to the psychological distress in families of FS patients and to the high prevalence (25%) of undiagnosed FS among the relatives. PMID- 9213383 TI - Cardiovascular involvement in relapsing polychondritis. AB - Relapsing polychondritis is an inflammatory disease that characteristically involves cartilagenous tissues. Cardiovascular involvement is a fairly common complication and the second most frequent cause of mortality in this disease. The case of a man with a progressive cardiac involvement, aortic incompetence, mitral regurgitation, and finally complete atrioventricular block offered the opportunity of reviewing the cardiovascular complications in relapsing polychondritis. The most frequent abnormalities are aortic regurgitation and aortic aneurysm. Furthermore, several cases of atrioventricular block, mitral regurgitation, and acute pericarditis have been reported. For early diagnosis and treatment of these severe complications, periodic cardiovascular examination is mandatory in these patients. PMID- 9213384 TI - Systemic lupus erythematosus and multiple myeloma: a rare association. AB - OBJECTIVE: The association of systemic lupus erythematosus (SLE) and multiple myeloma (MM) is an uncommon event. We report the relapse of SLE in a patient with a previous history of MM, treated with chemotherapy and, subsequently, with alpha 2b interferon (alpha-2b IFN) as a maintenance therapy. The case is discussed in light of past relevant literature. METHODS: The history and clinical, laboratory and radiographic findings of the patient, as well as the subsequent therapeutic approach are discussed. In our review of the literature, journal articles are identified by Medline search. RESULTS: We describe the case of a woman who developed a multiple myeloma 14 years after a diagnosis of SLE. A careful literature review confirms that the association of these two diseases has been reported only in a few cases. When the plasma cell neoplasia occurred, SLE had been quiescent for several years; the patient was treated with prednisone melphalan and, subsequently, with alpha-2b IFN as a maintenance therapy. On admission to our department, SLE was in a relapse phase, probably because of IFN treatment. The disease was poorly responsive to steroid therapy and required the use of cytotoxic drugs. CONCLUSIONS: The coexistence of SLE and MM is very rare and the possible pathogenetic mechanisms underlying this association remain unclear. The use of interferon in a patient with an autoimmune disease always invites caution. PMID- 9213385 TI - Blocking antibodies inhibit complete African swine fever virus neutralization. AB - A persistent non-neutralized African swine fever virus (ASFV) fraction is found with most convalescent swine sera in in vitro neutralization assays. To study this phenomenon, antisera from convalescent pigs infected with different virus isolates and showing complete or incomplete virus neutralization were used. Different experiments determined that incomplete neutralization of ASFV is caused neither by virus aggregation, nor low affinity or stability of virus-antibody complexes. Additionally, attempts to purify antigenic escape mutant viruses from the persistent fraction was also unsuccessful. Nevertheless, competition experiments between sera demonstrated that antibodies present in sera showing persistent fraction inhibited the complete neutralization mediated by antibodies present in sera which neutralize 100% of virus infectivity. These results suggest that induction of blocking antibodies during ASFV infection could represent the main cause for the persistent surviving virus fraction observed in neutralization assays and could also explain the persistent infections observed in some convalescent pigs. PMID- 9213386 TI - African swine fever virus-specific cytotoxic T lymphocytes recognize the 32 kDa immediate early protein (vp32). AB - African swine fever (ASF) virus-specific cytotoxic T lymphocyte (CTL) activity has been studied in a model in which SLA inbred minipigs were experimentally infected with an attenuated isolate of the virus. The CTL assays were performed using alveolar macrophages as target cells. The specific lysis is mediated by purified CD8+ lymphocytes but not by CD4+ cells and can be blocked by incubation with anti-SLA class I monoclonal antibodies. The purified CD8+ population produced high levels of interferon-gamma after ASF virus stimulation. In an attempt to define the viral proteins recognized by CTL, target cells infected with a recombinant vaccinia virus (VV) expressing the ASF virus p32, an immediate early protein during ASF virus replication, were recognized and lysed by CTL. This assay may be useful for VV recombinant screening in order to identify other potential target ASF virus proteins. PMID- 9213387 TI - Identification and comparison of point mutations associated in classic and variant infectious bursal disease viruses. AB - Restriction enzymes (RE) were used to identify point mutations in the nucleotide sequences of the infectious bursal disease virus (IBDV) strains Del-E, Del-A, STC, IN, Bursine 2, and Bio-Burs. The point mutations at amino acid sites 222, 254 and 323 were identified using REs BstNI, StyI and MboI, respectively. The nucleotide sequences of STC, IN, Bursine 2 and Bio-Burs were determined at each of these sites. Nucleotide sequence analysis using this data and previously reported data for Del-E and Del-A was used to confirm the point mutation and predict the resulting amino acids. Although there were some exceptions, the BstNI and StyI enzymes detected point mutations in the first base of the 222 and 254 codons, respectively and could be used to predict an amino acid change in the viruses. MboI was detecting a mutation in the third base of codon 323 and could not reliably predict an amino acid change at that position. The results indicated that amino acids 222 and 254 were consistently mutated in the variant viruses examined and that amino acid 323 was not. Furthermore, point mutations resulting in amino acid changes at position 222 suggested that several groups of viruses may be defined by this site alone; proline for classic strains like STC, threonine for Del-E and GLS, serine for IN, Bursine 2, and Bio-Burs and glutamine for Del-A. PMID- 9213388 TI - Experimental confirmation of recombination upstream of the S1 hypervariable region of infectious bronchitis virus. AB - Chimeric infectious bronchitis virus (IBV) genomes with cross-over sites in the S1 gene were generated by co-infection with two distinct IBV strains. Recombinant viruses were collected from chicken embryos, embryonic cultured cells and chickens co-infected with Ark99 and Mass41 strains and purified by differential centrifugation. The recombinant S1 genes were identified by reverse transcription polymerase chain reaction (RTPCR) using heterologous primers and confirmed by nucleotide sequencing. The recombinants with Ark99 5' and Mass41 3' sequences were identified following the in vitro, in ovo and in vivo co-infections. Mixed RNA extracted from Ark99 and Mass41 did not produce RTPCR products with these primers at the PCR conditions used. Cross-over sites within the amplified 580 (Mass41) or 604 (Ark99) bases of the 5' S1 gene could only be detected between nucleotides 50 and 155. While this region, lying upstream of the S1 hypervariable region, corresponded with sites commonly identified in naturally occurring isolates, recombination sites identified in these studies could not be detected within the HVR of S1 of the genomes of chimeric viruses. PMID- 9213389 TI - Analysis of the conformational change of recombinant human papilloma virus type 18 E7 protein induced by metal binding. AB - Human papillomavirus (HPV) type 18 E7 gene was isolated by polymerase chain reaction (PCR) amplification from tissues of Korean cervical cancer patients and cloned into a plasmid vector, pET-3a, for the expression of recombinant E7 protein (rE7) in Escherichia coli. The rE7 protein was purified to the homogeneity and its purity was confirmed by HPLC. The purified protein was analyzed for the metal-binding properties by UV spectroscopy and it was shown that two Cd2+ or Zn2+ ions bind to one E7 protein by the metal-sulfur ligand formation via two Cys-X-X-Cys motifs in E7 protein. When the change of intrinsic fluorescence of tryptophan residue was analyzed for rE7-Zn complex, the blue shift of emission wavelength and the decrease in maximum intensity of emission were observed compared with rE7. These results suggest that Zn(2+)-bound rE7 has undergone conformational change, in which a tryptophan residue located in the second Cys-X-X-Cys motif was moved into solvent-inaccessible or hydrophobic environment. The rE7-Zn complex was found to be resistant to chymotrypic digestion by comparing the digestion patterns of rE7. Therefore, we showed the folding status of HPV 18 E7 could be changed by metal binding resulting in a different conformation in which a tryptophan residue was driven into more hydrophobic environment and the resistancy to chymotryptic digestion was conferred. PMID- 9213390 TI - GB virus C (GBV-C)/hepatitis G virus (HGV) infection among intravenous drug users in Japan. AB - A sero-epidemiologic survey of 74 IVDUs and 86 age-matched controls was performed to investigate the prevalence, clinical significance, and genetic distribution of GB virus C (GBV-C)/hepatitis G virus (HGV) infection among intravenous drug users (IVDUs) in Japan. GBV-C/HGV RNA was detected by reverse transcriptase-hemi-nested polymerase chain reaction (RT-hemi-nested PCR) for the 5'-untranslated region (5' UTR) in 43.2% (32/79) of the IVDUs compared with 1.2% (1/86) of the controls (P < 0.001). The duration of drug use did not correlate with the prevalence of GBV C/HGV. Thirteen subjects positive for GBV-C/HGV RNA without HCV RNA had normal serum aminotransferase concentrations. Phylogenetic tree showed that the 32 GBV C/HGV-positive isolates from IVDUs were classified within a new GBV-C/HGV group, which has been identified mainly in Asian subjects (type 3). The present study illustrated, (1) the high incidence of GBV-C/HGV infection among Japanese IVDUs, (2) GBV-C/HGV alone may not be an important pathogenic factor for hepatic injury, and (3) type 3 GBV-C/HGV was the most prevalent among IVDUs in Japan. PMID- 9213391 TI - Self-association of truncated forms of HIV-1 gp120. AB - HIV-1 gp120 and truncated forms were expressed in HeLa T4 cells by vaccinia recombinant viruses. The truncated gp120 molecules consisted of N-terminal overlapping envelope proteins of 204, 287 and 393 amino acids respectively. Immunoprecipitation with specific monoclonal antibodies and SDS-PAGE analyses of HIV-1 gp120 revealed bands corresponding to low amounts of secreted and cell bound stable dimers. In contrast, the truncated forms of gp120 expressed larger amounts of SDS-stable putative dimers and the amounts observed were inversely proportional to their size. The shortest gp120 mutant (204 aa) was found to be secreted almost exclusively as a dimer. The processing of gp120 and its truncated forms was further investigated in the presence of inhibitors of N-glycosylation. Monomers and dimers migrated on gels with the same relative changes, confirming that the protein with the higher molecular weight is a multimer of the smaller one. The putative dimeric form of the truncated gp120s could be stabilized by chemical cross-linking. Finally, the possible existence of an association domain in the N-terminal 204 amino acids (aa) of gp120 is discussed. PMID- 9213392 TI - Virulence-associated sequence duplication at the hemagglutinin cleavage site of avian influenza viruses. AB - Recent highly pathogenic (HP) field isolates of avian influenza (AI) virus from Mexico all possess an insertion of at least two basic amino acids (arg-lys) at the cleavage site of the hemagglutinin (HA) glycoprotein. One HP isolate has additional information which yields a 4 amino acid insert (arg-lys-arg-lys). We present here the nucleotide sequence of the HA gene of this unique isolate and compare it to recent H5N2 and other avian influenza isolates. The complete HA nucleotide sequence of the isolate and phylogenetic relationship suggest that it was derived in direct succession from a non-pathogenic strain isolated about 1 month earlier. The unique insertion sequence is a direct duplication of part of the purine-rich region preceding the arginine codon at the HA cleavage site. This evidence along with other data in this report provide compelling support for a proposed model explaining the mechanism of spontaneous, virulence-related insertions in type A influenza viruses. PMID- 9213393 TI - Sequence variation in the early genes E1E4, E6 and E7 of human papilloma virus type 6. AB - The majority of condylomata acuminata (anogenital warts) are caused by infection with Human Papilloma Virus type 6 (HPV-6). We have sequenced the HPV-6 early genes, E1-E4, E6 and E7 from wart biopsy DNA samples sourced from the UK and USA and derived a consensus sequence for these genes and the proteins they encode. When compared to the prototype HPV-6b sequence, published over 12 years ago, the E1-E4 consensus sequence showed 3/91 (3.3%) amino acid changes, the E6 consensus sequence showed 1/150 (0.7%) changes and the E7 consensus sequence showed 1/98 (1.0%) changes. Since many of the early gene sequences from biopsy material were more similar to the HPV-6a subtype than HPV-6b, this data supports the use of HPV 6a as the HPV-6 prototype. PMID- 9213394 TI - Protein cell receptors mediate the saturable interaction of African swine fever virus attachment protein p12 with the surface of permissive cells. AB - Previous studies have demonstrated that the entry of African swine fever virus (ASFV) into Vero cells and swine macrophages is mediated by saturable binding sites located on the plasma membrane. The ASFV protein p12 has been implicated in virus attachment to the host cell, but the cellular component responsible for the interaction with the virus is largely unknown. We have studied the binding of recombinant p12 and ASFV to different cell lines. Permissive cells were able to bind p12 in saturable and nonsaturable interactions, as reported for ASFV. Experiments of binding recombinant p12 have been used for the initial characterization of the specific receptors on Vero cells. The treatment of cell surfaces with different enzymes and lectins resulted in the inhibition of the p12 binding activity by several proteases, but not by glycosidases or lipase, suggesting that the receptor is composed of protein, with no carbohydrates or lipids involved in the virus attachment to the cellular membrane. The recovery of receptor activity after pronase treatment was completed in 6 h in culture medium containing tunicamycin, and could not be restored in the presence of cycloheximide, confirming that synthesis of new proteins, but not glycosylation, was required for the recovery of the receptor activity. These data support the idea that membrane protein(s) on the surface of permissive cells act as receptors for ASFV and that this specific interaction is, at least, one necessary step in a productive virus infection. PMID- 9213395 TI - Simian immunodeficiency viruses containing mutations in the long terminal repeat NF-kappa B or Sp1 binding sites replicate efficiently in T cells and PHA stimulated PBMCs. AB - The long terminal repeats (LTRs) of primate lentiviruses contain conserved binding sites for the NF-kappa B and Sp1 cellular transcription factors. In order to study the role that these sites play in simian immunodeficiency virus (SIV) replication, we have introduced mutations that disrupt either the NF-kappa B or Sp1 binding sites in the LTR of an infectious molecular clone of SIVmac239. An additional mutation also disrupted the SF3 transcription factor binding site that overlaps the NF-kappa B site. Viruses containing point mutations or deletions of the NF-kappa B, SF3, or Sp1 binding sites retained the ability to replicate efficiently in the CEMx174 and MT4 cell lines, as well as in PHA-stimulated primary rhesus macaque peripheral blood mononuclear cells (PBMCs). Efficient replication of SIVs mutated in either NF-kappa B or Sp1 binding sites suggests that the SIV LTR promoter contains multiple functionally redundant elements capable of supporting sufficient transcription to allow productive viral replication. PMID- 9213396 TI - Specific binding of HIV-1 envelope protein gp120 to the structural membrane proteins ezrin and moesin. AB - The observation that HIV in vitro can infect CD4- and Gal-C-negative brain cell lines has stimulated this study to identify alternative gp120-binding proteins on brain cells. HIV-1 gp120 binding proteins of the CD4-negative and Gal-C-negative, non-productively infectable human glioblastoma cell line D54 were purified by affinity chromatography over a gp120-conjugated sepharose column and identified by peptide microsequencing. The binding capacity and specificity of this column was controlled using extracts of CD4-positive cells. Two of seven prominent proteins eluted from the gp120 affinity column specifically bound gp120 in Western blot overlay experiments and were identified by subsequent immunoblotting and microsequencing as ezrin and moesin, members of the ERM (ezrin, radixin, moesin) family of cellular structural membrane proteins. Antibodies to ezrin and moesin specifically recognized the eluted gp120 binding proteins confirming their identification. Ezrin and moesin are structural proteins binding to the cellular membrane and to several cytoskeletal and transmembrane proteins. Our results suggest that ezrin and moesin might play a role as gp160/gp120 binding proteins during the uptake, the assembly or the budding of HIV. PMID- 9213397 TI - [Repair of a large nasal septum perforation]. PMID- 9213398 TI - [Progressive hearing loss in children with hearing aids]. AB - BACKGROUND: The possibility of hearing aid induced deterioration was first described in 1939 (Berry 1939, Holmgren 1939). Since then numerous studies discussed this problem controversely. However, it is undisputed that in cases of profound sensorineural hearing loss in childhood powerful hearing aids are necessary for maturation and auditory and speech development. An increase of progressive sensorineural hearing loss in the last three years in children with profound sensorineural hearing loss fitted with hearing aids (2 children in 1993, 6 children in 1996) drew our attention to this problem. METHOD: Over a period of four years, we collected the data of 16 children with progressive hearing loss. RESULTS: The comparison of the duration of the hearing aid use since fitting and the maximum output level fail to explain this increase. Moreover our strategy in hearing aid fitting remained unchanged during this period especially with regard to SSPL max. We also performed an analysis of the childhood disorders associated with the sensorineural hearing losses. This analysis provided no further information for the increase of hearing loss. CONCLUSION: There is no evidence that powerful hearing aids damage hearing and induce progressive sensorineural hearing loss in childhood. Therefore we believe that there is no need to change our hearing aid fitting strategy particularly with regard to the maximum output level up to 135 dB SPL measured in situ. The increase in the number of progressive cases in the last two years indicates the necessity of further investigations in this field. PMID- 9213399 TI - [Microcirculation of the nasal mucosa during use of balloon tamponade]. AB - BACKGROUND: Nasal packings are commonly accepted in the treatment of severe epistaxis. Cuffed catheters are known to cause damage to the nasal mucosa most likely by interfering with tissue perfusion. In this study the effect of different pressure levels on local perfusion of septal mucosa is investigated. METHOD: In 15 healthy subjects the blood flow in septal mucosa was measured by laser doppler flowmetry by positioning a cuffed epistaxis catheter into the nasal cavity with a laser probe attached to it. Increasing pressure was administered by injecting saline solution while continuously recording intraluminal pressure, perfusion, and filling volume. The local pressure affecting the septal mucosa at the moment of stalling perfusion was determined by subtracting the extranasal cuff pressure from the current intranasal cuff pressure at same inflation volumes. RESULTS: Microcirculation of the septal mucosa stopped when the local pressure exceeded a value of Pmean = 42 mmHg. Individual variations (n = 15) were small (s = 9 mmHg). The intraluminal cuff pressure was measured to be about ten times higher due to the retraction force of the cuff. Spontaneous oscillations of the blood flow were reduced with increasing pressure to the blood vessels. Filling volumes up to 3.2 ml were sufficient to stop perfusion. CONCLUSIONS: Cuffed nasal packings stop the blood flow in nasal mucosa even at low local pressures. Depending on the material characteristics of different cuffs the pressure to dilate the cuff may, however, be several times higher than the actual local pressure. This effect may cause problems in the proper use of cuffed catheters. Laser doppler flowmetry proved to be helpful in determining reproducible perfusion values. PMID- 9213400 TI - [Endonasal, microscopically controlled frontal sinus surgery]. AB - BACKGROUND: The anatomic variation of the frontal sinus and frontal recess can create both a diagnostic and therapeutic challenge. Most cases of frontal sinus disease can now be treated by endoscopic approaches. For refractory cases or those with severe pathology, the microscopically controlled drainage (MCD) operation has at times been successful and spared the patient the morbidity of an external approach. The aim of this study was to evaluate microscopically controlled frontal sinus surgery in these difficult cases. MATERIAL AND METHODS: Prospective analysis was performed on the efficacy of MCD in patients for whom endoscopic sinus surgery had failed or in primary cases with severe pathology or unfavorable anatomy. The technique employs a self-retaining endonasal retractor and a diamond bur under microscopic visualization to remove solid bone (frontal spine) obstructing the sinus drainage and allow a wide opening of the frontal recess while causing minimal mucosal damage. Unilateral drainage (extended frontal sinus drainage operation), and in some cases bilateral drainage (median drainage procedure) is employed. RESULTS: With an average of 23 months of follow up, over 90% of patients were either free of symptoms or substantially improved after the MCD procedure. Three patients required revision surgery (extend the opening into a median drainage procedure) for adequate relief of symptoms. CONCLUSIONS: The MCD procedure is highly successful in the treatment of frontal recess disease, particularly in those cases of severe pathology or difficult anatomy. It may be used in those cases refractory to standard endoscopic sinus surgery where an external approach and frontal sinus obliteration are contemplated. As with endoscopic sinus surgery, precise knowledge of the frontal recess and neighboring landmarks is critical, and adequate drainage with minimal mucosal disruption should be the goal. PMID- 9213401 TI - [Effectiveness and tolerance of nasal irrigation following paranasal sinus surgery]. AB - BACKGROUND: Cleaning of the nose with saline solution after endonasal sinus surgery is very often used for postoperative treatment. But the efficiency and acceptance of this method has not been examined thoroughly until now. METHODS: Performance, effectiveness and acceptance of the postoperative treatment was evaluated in a questionnaire. One hundred thirty-four of 180 patients answered. RESULTS: One hundred twenty-one patients (66.1%) cleaned their nose with Ems brine; 28.1% of the patients used saline solution. The nasal douche was the cleaning device used by 39.7% of the patients, whereas 53.7% sniffed the solution from their hand. Ninety-five percent found that this kind of treatment was easy to do; 84.7% found it equally pleasant. There was no difference between using Ems brine or NaCl solution nor between using the nasal douche or sniffing out of the hand. Fifty-one point four percent of the patients with an follow-up of 27 to 36 months rinsed their nose up to now; 55.9% resumed nasal irrigation after an interval. CONCLUSIONS: Rinsing of the nose after endonasal sinus surgery is judged positively by most of our patients and is integrated well in the daily routine. Although it is most common to sniff saline solution out of the hand, existing research recommends usage of warm Ems brine in combination with the nasal douche. PMID- 9213402 TI - [High-risk HPV types in oral and laryngeal papilloma and leukoplakia]. AB - BACKGROUND: Juvenile and adult laryngeal papillomas (JLP and ALP) and oral papillomas (OP) are important benign tumors of the head and neck. Laryngeal leukoplakia (LL) may be a precancerous lesion. The etiology of the papillomas is associated with human papillomavirus infection (HPV). The important noxes for the development of laryngeal leukoplakias are nicotine, alcohol, and HPV. Adult laryngeal papillomas and OP can also undergo malignant conversion. Today, there is no marker known to distinguish in progressive lesions and in those which show a regression. Different types of HPV were detected in head and neck cancer too. There is a important similarity to the genesis of cervical cancer. The 77 known HPV types were divided into benign types, e.g., 6 and 11, and those with oncogene potential, e.g., 16, 18, 31, 33, and 35. The detection of oncogene HPV may be a sensitive marker for prognosis of primary benign lesions. PATIENTS AND METHODS: In this study, the presence of HPV genomes 6, 11, 16, 18, 31, 33, and 35 in 17 JLP, 27 ALP, 15 OP, and 11 LL was examined. DNA extracted from archived samples embedded in paraffin was amplified using the E6 specific polymerase chain reaction (PCR). The products were visualized by electrophoresis, and positive identification was achieved by Southern blot analysis and hybridization to specific biotinylated oligonucleotide. RESULTS: Our data show the presence of HPV 6/11 in all JLP (17 of 17), in all ALP (27 of 27), in 13 of 15 (87%) OP, and in seven of 11 (63%) LL. The "malignant" types HPV 16, 18, and 33 were found in six of 27 (22%) of the ALP, in three of 15 (20%) of the OP, and in four of 11 (36%) of the LL. the dominant type was HPV 16, HPV 31 and 35 were not detectable. Three ALP, one OP, and the four LL of the cases with oncogene HPV showed histologic features of moderate dysplasia. CONCLUSIONS: The role of HPV in malignant transformation of infected cells remains unclear. It is well known that the carcinogenesis must depend on promoters such as alcohol, tobacco, and metabolites of chronic inflammations. All patients with positive biopsies confirming HPV 16, 18, or 33 must receive special care to prevent the development of a carcinoma. PMID- 9213403 TI - [Malignant degeneration of juvenile laryngeal papillomatosis?]. AB - BACKGROUND: Spontaneous malignant transformation of laryngeal papillomatosis was in the past mostly negated, or the discussion in literature was rather toned down and reserved. Therefore, from a biological and prognostic point of view, HPV infection of the larynx seems to carry a different weight than a viral infection in genital region. According to general consensus, secondary, malignant transformation in juvenile papillomatosis occurs in irradiated patients and leads to the conclusion that radiation therapy of this disease is presently contraindicated. Because there is as yet no causal and curative treatment, repeated and frequent removal of papillomatous tissue by microlaryngoscopy may often be necessary to keep the airway patent. PATIENT, METHOD AND RESULTS: We diagnosed and treated an advanced laryngeal squamous cell carcinoma with lymphatic metastasis in a 50-year old male. Juvenile papillomatosis had been diagnosed already at the age of five, and at the patient's last presentation 5 years ago (age 45), typical clinical and histological features of laryngeal papillomatosis had been observed. Furthermore, virus infection of the papillomatous tissue (HPV-6/11) was proved by using the technique of in-situ hybridisation. Risk factors for malignant transformation, such as smoking, alcohol or radiation, were denied by the patient. CONCLUSIONS: From these aspects, a spontaneous, malignant transformation of laryngeal papillomatosis must be considered with regard to six similar observations in the German and English literature. In the reported case, a tumoural origin in the flat laryngeal mucosa in close neighbourhood to the former site of papillomas, is less probable, albeit not ruled out completely, since continuous changes from benign squamous papilloma to atypical, invasive tumour and a HPV-infection in the carcinomatous tissue could not be proved by in-situ hybridisation. PMID- 9213404 TI - [Psychosocial aspects in total external ear reconstruction in patients with severe microtia]. AB - BACKGROUND: The aim of this study was to identify the psychosocial problems and expectation of patients with severe microtia and to investigate possible changes in psychosocial profile of patients after surgical reconstruction of the pinna. METHODS: Interviews were performed with 65 patients before and 39 patients after auricular reconstruction. They consisted of a questionnaire involving open and closed questions relating to the reconstruction and a personality test. RESULTS: The most frequently mentioned reasons for surgery were: personal dissatisfaction with patients own appearance, lack of self-confidence, and for younger age groups, parental anxiety about the future psychosocial development of their children. More than 90% of the patients with surgically reconstructed auricles were satisfied with the postoperative results and showed a more open and fearless personality profile. CONCLUSIONS: Auricular reconstruction enhances the development of a stable personality. PMID- 9213405 TI - [Clinical comparison between radial and lateral forearm flap]. AB - BACKGROUND: Free fasciocutaneous flap transplantation is a versatile method for soft tissue reconstruction. This clinical study points out differences between the radial forearm flap and the lateral arm flap. METHODS: We used the radial forearm flap in 36 patients following tumor ablation and in 11 patients we used the lateral arm flap for soft tissue reconstruction. We studied the arterial and venous vessel calibers of the flaps, the vessel pedicle length, and the size of the skin paddle. Motor and sensory function tests of the upper/ lower arm and hand were performed after surgery. Recipient and donor site morbidity was noted. RESULTS: Compared to the forearm flap the lateral arm flap is bulky (1-5 cm vs. 0.5-1.5 cm), its vessel calibers are smaller (Art.: 1.4 vs. 1.8 mm, Ven.: 1.8 vs. 2.0 mm), flap size and maximum vessel pedicle length (10 vs. 12 cm) are equal. Raising the lateral arm flap is more demanding and needs more time due to the deep location of the vessel pedicle and the accompanying radial nerve within the intermuscular septum. On the other hand the lateral arm flap is advantageous because of primary wound closure of the donor site. The donor site of the forearm flap had to be covered with skin graft in all cases. We found sensory deficits of the proximal lower arm in 50% after dissection of the lateral arm flap and in 14% on the distal lower arm and thumb joint after dissection of the radial forearm flap. CONCLUSIONS: Both transplants are fasciocutaneous and optional innervated, they offer a constant anatomy and can be harvested simultaneously without interference to the head and neck team. Because of the specific characteristics of these flaps we prefer the radial forearm flap for soft tissue reconstruction. We use the lateral upper arm flap, if a forearm flap cannot be harvested, for head or neck augmentation and for reconstruction of large and deep defects. PMID- 9213406 TI - [Plastic reconstructive surgery of soft palate defects--functional and oncological aspects]. AB - BACKGROUND: Resection of the soft palate in tumor surgery can lead to significant swallowing disorders. Therefore rehabilitation needs a functional reconstruction of the remaining defects. MATERIALS AND METHODS: In four years, nine patients received a partial removal, and six patients a total removal of the soft palate including adjacent parts of the oropharynx in six cases due to oropharyngeal carcinoma. Partial defects were reconstructed with a neurovascularized infrahyoidal muscle flap, total defects with fasciocutaneous flaps of the lateral arm, radial forearm or scapula region. The function of the new palate was evaluated by interview, cinematography, and pressure measurements of the pharynx. RESULTS: These investigations demonstrated proper swallowing without aspiration or regurgitation in all cases. Values of 60% of normal pressure behind the palate have been achieved after palate reconstruction with a pressure slope directed into the hypopharynx. Decannulation was possible on average 34 days postoperatively, removal of the feeding tube on average 29 days postoperatively. Now 87% of our patients are free of tumor after a mean observation time of 24 months. In light of the fact that two-thirds of all patients suffered from advanced carcinoma, results can be considered as good. This study shows good functional and oncologic results after tissue reconstruction of the soft palate. PMID- 9213407 TI - [Perfusion manometry in the evaluation of postoperative swallowing function following various reconstructive procedures of the upper aero-digestive tract]. AB - BACKGROUND: The swallowing function can be restored after large oncologic resections in the upper aerodigestive tract with microvascular anastomosed transplants. PATIENTS AND METHODS: With the help of the perfusion manometry we would like to demonstrate the functional advantages of this reconstruction method. We examined three reconstructed regions: tongue with 15 patients, soft palate with 11 patients, and the laryngeal and pharyngeal complex after total laryngopharyngectomy with 17 patients. RESULTS: Patients with reconstructed tongue or soft palate reached 69% or 74% of their pressure compared to normal values. CONCLUSIONS: We demonstrated that although normal pressure values were not reached after reconstruction of large defects with microvascular anastomosed transplants; these reconstruction methods restored the pressure gradient essential for swallowing, with a higher pressure of the soft palate and a lower pressure of the base of tongue. PMID- 9213408 TI - [Icelandic moss lozenges in the prevention or treatment of oral mucosa irritation and dried out throat mucosa]. AB - BACKGROUND: Icelandic Moss (Cetraria islandica) has been used to treat inflammation and dryness of the pharyngeal mucosa in the naturopathy for many years. It contains substances with protective and antiinflammatory effects for the oral mucosa. The necessary concentration of Icelandic Moss and its therapeutic effectiveness were analysed in this clinical study. METHODS: Sixty one patients who recently underwent surgery of nasal septum were evaluated for this randomized study. These patients suffer especially from dryness and inflammation of the oral cavity due to breathing only through the mouth while the nose is permanently closed by a nasal package. Three groups of patients treated with three different concentrations of the active substance were compared, Coating, dryness, and inflammation of the mucosa, lymph nodes, tongue, the tolerance of the drug, and symptoms like hoarseness and sore throat were documented. The oral application was performed from the first to the fifth day after the operation. RESULTS: The treatment with Icelandic Moss caused a direct reduction of the observed pathological changes. There were no significant differences between the three test groups. A dose of 0.48 g per day (10 Isla-Moos lozenges per day) is a sufficient treatment. CONCLUSIONS: Icelandic Moss is an advisable therapeutic opportunity without interactions and side effects for the treatment of inflammation of the oral mucosa occurring after nasal surgery, after intubation, and for simple infections of the throat. PMID- 9213409 TI - [Significance of p53, PCNA and Ki-67 in the prognosis of squamous cell carcinoma of the oral cavity]. AB - BACKGROUND: A common gene alteration in human malignancies is the mutation of the p53 gene. Through mutation p53 loses its function as a tumor suppressor and enables the tumor to proliferate. Ki-67 and PCNA are proliferation markers. The aim of this study was to find a reference to the prognostic significance of p53, Ki-67, and PCNA in squamous cell carcinoma (SCC) of the oral cavity. MATERIAL AND METHODS: Samples from 64 patients of the Department of Otolaryngology/Head and Neck Surgery, University of Bergen, Norway, with histologically proven SCC of the oral cavity were submitted to immunohistochemical procedure. All carcinomas were untreated. RESULTS: 59.3% of the patients showed a positive reaction for p53.Ki 67 and PCNA were expressed in about 68% of the cases, but with less staining intensity. No correlation could be found between the expression of the three antibodies and tumor factors such as tumor size, lymph nodes, degree of differentiation, recurrence and survival rate, the occurrence of second primaries, or the smoking habits of the patient. CONCLUSIONS: The expression of the tumor suppressor gene p53 and the proliferation marker PCNA and Ki-67 seem to have no prognostic significance in SCC of the oral cavity. PMID- 9213410 TI - Enzymatic and electrochemical reactions of xanthine oxidase immobilized on carbon materials. AB - An optimum way of immobilizing xanthine oxidase on graphite was found where a redox transformation of the enzyme was observed. The nature of the redox maxima was hypothesized on the basis of the dependence of the half-wave potential (Ep/2) on the pH of the solution. The enzymatic activity of xanthine oxidase adsorbed on two kinds of soot was studied by the oxidation of xanthine. The kinetic and activation parameters of the enzyme reaction were determined. PMID- 9213411 TI - Role of hydrophobic and hydrophilic interactions of organotin and organolead compounds with model lipid membranes. AB - The present study was conducted to clarify the mechanism of toxicity of organic compounds using lipid model membranes (liposomes and planar lipid membranes). The compounds studied were trialkyltin and trialkyllead chlorides, dialkyltin dichlorides and some inorganic forms of those metals. Two different (anionic and cationic) detergents were also used in the experiments to change the surface properties of liposomes. As a measure of interaction between the compounds studied and model membranes were the release of liposome bound praseodymium and the change in stability of planar membranes under the influence of those compounds. On the basis of the results obtained it was postulated that the mechanism of interaction between tin- and leadorganics and model lipid membranes is a combination of different factors featuring interacting sides. The most important properties determining the behaviour of organic compounds in the interaction were lipophilicity and polarity of different parts of the organics and the steric arrangement they can take in the medium. On the other hand, the surface potential of the lipid bilayer and the environment of the lipid molecules, that play a significant role in the availability of the lipid bilayer to the organics, were important factors in the interaction. PMID- 9213413 TI - AAMC project committee on increasing women's leadership in academic medicine. PMID- 9213412 TI - Stimulation of pancreatic lipase activity by saponins isolated from Medicago sativa L. AB - Saponins isolated from Medicago sativa L. appeared to stimulate lipolytic activity and did not influence amylolytic or proteolytic activity of Neopancreatinum (extract of pancreatic enzymes: lipase, proteases and amylases). Saponins isolated from the aerial part of Medicago sativa L. were demonstrated to stimulate lipase activity more effectively than saponins separated from the root of this plant. When saponins were treated with 0.02 M HCl at 37 degrees C for 1 hour their lipase stimulation ability did not change. The saponins stimulated Neopancreatinum lipolytic activity more intensively in the presence of sodium cholate. PMID- 9213414 TI - [Use of flow cytometry for HLA B27 phenotyping: study of a HLA B7/HLA B27 double marker technique]. AB - Among HLA-associated diseases, the strong association between HLA B27 and ankylosing spondylitis (AS) allows to perform HLA B27 typing for the diagnosis of this disease. The technique described: double immunostaining anti-HLA B7/HLA B27 on whole blood samples and analysis by flow cytometry is a rapid method for the detection of HLA B27 antigen. We have compared the results obtained with those of the TERASAKI microlymphocytotoxicity reference method on a population of 300 patients. The absence of false negative results indicates that this test is suitable for AS screening. The use of two monoclonal antibodies proves to be effective in eliminating cross reactions described with HLA B7 flow cytometric techniques using single monoclonal antibody. This technique allows to restrict the use of microlymphocy-totoxicity to patients presenting with both HLA B7 and B27 positive reactivities. PMID- 9213415 TI - [Contribution of flow cytometry to allergologic diagnosis]. AB - The technique of Flow Cytometry for activation of basophils (TAB), expressed as the marker CD63, is at present one of the pathways of research applied to drug allergy. In this work are reported the results of TAB of Pneumoallergens and Hymenoptera venoms. TAB can define better than total IgE the atopy of the subject: in effect the fluorescence of the basophils is emphasized in comparison with non-atopic subjects. This hypothesis has been confirmed by a study of three groups of subjects. With regard to drug allergy, it is important to study patients in whom the observations have been documented very objectively by clinical history and positive skin tests to the drug, compared with a negative reference to the same drug. So, TAB has been shown to be very useful in diagnosis of allergy to certain drugs, such as the Myorelaxants. PMID- 9213416 TI - [Chronic rhinitis, or the consequences of gluttony]. PMID- 9213417 TI - [Physiopathology of food allergy]. AB - Food allergy is defined by a mixture of digestive and or extra-digestive clinical symptoms, "rhinitis, asthma, eczema, etc" secondary to contact with a food and/or a preservative and/or addition to the immune system of the organism, with its origin in an excessive humoral and/or cellular response, called HYPERSENSITIVITY. For this to occur, food allergy needs conjunction of several parameters. Predisposing factors Genetics, with facility to produce IgE. Reduction of the defences of the digestive system. Usual foods PMID- 9213419 TI - [Allergy to cow's milk]. AB - After recalling the medical reluctance as well as the risks that there are in complete elimination of milk in infants, the author presents several clinical pictures and then a classification of the immunological types: Allergic shock of neonates, digestive and extra-digestive (skin and respiratory airways) symptoms finally the rare chronic gastro-enteritis to cow milk. Non-reaginic food allergies: Acute gastro-enteropathy to cow milk, with villous atrophy and Heiner's syndrome, delayed hypersensitivities are studied, of difficult diagnosis that may cover almost all pathologies. They may be found in the digestive system, respiratory, the kidneys and even in the organs of behaviour. Migraine of food origin must be remembered. Development in regressive rules is a function of the type of allergy and the suddenness of the symptoms. Diagnosis is above all by questioning and confirmation or not by skin and in vitro tests. Certainty can only be shown by tests of elimination and re-introduction. The diet, at the same time of both diagnostic and therapeutic value, is based on the replacement of cow milk by foods that contain the same amount of proteins. It is essential, especially in the very small, to have perfect match of food so as to avoid any risk of a dramatic hypoprotinemia, which may happen if the child does not like the suggested diet, or if the parents cannot buy the substitution products. In such conditions great care must be taken to avoid provoking a crisis. Care must be taken to decide: If the elimination of cow milk is always justified each time. If it is, always check that the substituted protein is properly made, the family may change the diet mistakenly. PMID- 9213418 TI - [Immunobiologic diagnosis of food allergy]. AB - Food allergy is becoming more frequent, with 6% of asthmatics reporting an isolated food allergy, and 5 to 6% of atopic dermatitis patients also have either a single or multiple true food allergy. There is value in immuno-biological diagnosis by: Measurement of total serum IgE. Measurement of mono-allergen specific IgE, following a measurement by a multi-allergen of the Trophatope type. A study of elimination of foods for 2 or 3 months followed by their re introduction. Oral provocation tests in a hospital environment under clinical control and subsequent measurement of the mediators: Plasma histamine, tryptase, and urinary methylhistamine, to give proof of responsibility of the food allergen. Nowadays, it is perfectly possible to include in diagnosis the new technologies of the test of activation of basophils/or lymphocytes by means of flow cytometry. PMID- 9213420 TI - [Histamine liberation in vitro or histamine release (HR). Study of the threshold of positivity in allergy to hymenopteran venoms]. AB - The technique of histamine release in vitro (HR) is only usable in diagnosis of hymenoptera venom allergy. The allergen extracts that are supplied for clinical (skin tests) or biological diagnosis contain significant amounts of histamine, falsify the results, especially HR, and reduce significantly the correlation between HR and other biological or cutaneous tests. Dialysis of the allergen extracts significantly improves the sensitivity and specificity of the technique and lowers the threshold of positivity of HR from 30% to 12%. Correlation of HR with other diagnostic parameters:clinical history, specific IgE and skin tests is very significantly improved. PMID- 9213421 TI - [Factors associated with convalescence and role reprisal in women after a coronary artery bypass]. AB - The goal of this study was to describe how psychosocial, cultural, and sociodemographic factors influence the convalescent phase of women who have had an aorto coronary bypass graft (CABG) and to explore the relationship between those factors, the perception of health, and the recovery process. The women participating in this study were recruited during the preoperative period and contacted during the 9th and 12th week post surgery. It was observed that these women had psychosocial difficulties in the preoperative period. The study results demonstrated that some of those factors were influencing their health perception during the preoperative period. There did not appear to be any relation between their health perception and their recovery. Additional analysis demonstrated that their health perception in the postoperative period was better than during the preoperative period, and that their status improved according to the New York Heart Association Classification. Recommendations for future research and nursing practice are proposed. PMID- 9213422 TI - Glossopharyngeal neuralgia with tongue carcinoma. PMID- 9213424 TI - Merck Frosst award lecture 1995. La conference Merck Frosst 1995. Structural studies of lipoproteins and their apolipoprotein components. AB - Lipid transport processes via the circulatory system of animals are a vital function that utilizes highly specialized lipoprotein complexes. These complexes of protein and lipid impart solubility to otherwise insoluble lipids. The apoprotein components of lipoprotein complexes serve to stabilize the lipid components and modulate particle metabolism and function as ligands for receptor mediated endocytosis of lipoproteins. We have used an insect (Manduca sexta) model system for studies of lipid transport. In this system, flight activity elicits a dramatic increase in the demand for glycerolipid fuel molecules by flight muscle tissue. These lipids are mobilized from a storage organ and transported through the hemolymph (blood) to the flight muscle by the lipoprotein, lipophorin. This system possesses the unique property that lipids are loaded onto pre-existing high density lipophorin through the action of a lipid transfer particle (LTP). LTP is a high molecular weight hemolymph component that facilitates net vectorial lipid transfer from fat body tissue to lipophorin. The increase in lipid content of the lipoprotein induces association of a low molecular weight amphipathic exchangeable apolipoprotein, apolipophorin III (apoLp-III). ApoLp-III is a 18 kDa protein that normally exists as a water soluble monomeric hemolymph protein. The structural properties of apoLp-III have been investigated by X-ray crystallography. ApoLp-III from Locusta migratoria adopts a five helix bundle conformation wherein each of the amphipathic helices orients with its hydrophobic face directed toward the interior of the bundle. It has been hypothesized that lipid association requires a dramatic conformational change wherein the helix bundle opens about putative hinge domains located in the loops between helices. The data accumulated support the concept that apoLp-III is a member of the broad class of exchangeable apolipoproteins and structural information learned from this system is directly applicable to analogous proteins in higher organisms. PMID- 9213423 TI - Boehringer Mannheim award lecture 1995. La conference Boehringer Mannheim 1995. De novo design of alpha-helical proteins: basic research to medical applications. AB - The two-stranded alpha-helical coiled-coil is a universal dimerization domain used by nature in a diverse group of proteins. The simplicity of the coiled-coil structure makes it an ideal model system to use in understanding the fundamentals of protein folding and stability and in testing the principles of de novo design. The issues that must be addressed in the de novo design of coiled-coils for use in research and medical applications are (i) controlling parallel versus antiparallel orientation of the polypeptide chains, (ii) controlling the number of helical strands in the assembly (iii) maximizing stability of homodimers or heterodimers in the shortest possible chain length that may require the engineering of covalent constraints, and (iv) the ability to have selective heterodimerization without homodimerization, which requires a balancing of selectivity versus affinity of the dimerization strands. Examples of our initial inroads in using this de novo design motif in various applications include: heterodimer technology for the detection and purification of recombinant peptides and proteins; a universal dimerization domain for biosensors; a two-stage targeting and delivery system; and coiled-coils as templates for combinatorial helical libraries for basic research and drug discovery and as synthetic carrier molecules. The universality of this dimerization motif in nature suggests an endless number of possibilities for its use in de novo design, limited only by the creativity of peptide-protein engineers. PMID- 9213425 TI - Basement membrane induced differentiation of HEC-1B(L) endometrial adenocarcinoma cells affects both morphology and gene expression. AB - In vitro studies of endometrial carcinogenesis have been hampered by dedifferentiation of the cells in culture. Using the endometrial carcinoma cell line HEC-1B(L), we aimed to establish and characterize culture conditions that preserve a more differentiated state of the tumor cells. HEC-1B(L) cells grown in a serum-free defined medium on plastic (PL/SFDM) on top of a reconstituted basement membrane (Matrigel, MG/SFDM) or in a thick layer of Matrigel showed pronounced morphological differentiation as compared with HEC-1B(L) cells cultured on plastic in a medium containing serum (PL/10% FCS). Features of differentiation included cuboidal to columnar cell shape and an increase of rough endoplastic reticulum in Matrigel cultures. Gene expression of HEC-1B(L) cells was studied by metabolic [35S]methionine labeling and SDS-gel electrophoresis. HEC-1B(L) cells cultured in the presence of Matrigel showed two additional secretory proteins approximately 31 kD and 77 kD in size. rt-PCR was used to screen cell cultures for the presence of estrogen receptor, progesterone receptor, and lactoferrin-mRNA, genes typically expressed by normal endometrial epithelium. We found no expression of the estrogen receptor or progesterone receptor. Lactoferrin-mRNA was present under all culture conditions tested. Our results suggest a regulatory role of the extracellular matrix for the differentiation of the HEC-1B(L) cell line. PMID- 9213426 TI - Developmental expression of the collagen-binding heat-shock protein GP46 and collagen types I and IV in rat tissues. AB - The temporal expression of protein and mRNA encoding the collagen-binding heat shock glycoprotein, gp46, were determined in the heart, kidney, and lung during early rat postnatal development. The steady-state levels of collagen types I and IV mRNA expression were also examined to determine if gp46 and these collagen types are co-regulated during ontogenesis. Western blot analysis using a monoclonal antibody to gp46 revealed that gp46 levels are developmentally regulated. In heart and kidney, gp46 levels were high on days 3 and 8, reduced significantly on day 25, and low to undetectable on day 69. Protein levels of gp46 in the lung exhibited a similar temporal pattern except on day 3, when very low levels of gp46 were detected. mRNA expression of gp46 during early postnatal development did not correlate with gp46 protein accumulation in these tissues, suggesting a complex pre- and post-translational regulatory scheme. In the heart, protein levels of gp46 correlated well with collagen type I alpha 1(I) mRNA expression but not with collagen type IV alpha 1(IV). In contrast, gp46 protein levels closely paralleled alpha 1(IV) expression in the kidney. Gp46 levels exhibited no apparent correlation with either alpha 1(I) or alpha 1(IV) levels in the lung. These results show that gp46 is developmentally regulated at both the protein and mRNA levels in a tissue specific manner. The relationship between gp46 and collagen alpha 1(I) and alpha 1(IV) chain mRNA expression also has been shown to be tissue specific. PMID- 9213427 TI - Expression and epitopic conservation of calponin in different smooth muscles and during development. AB - Calponin is a thin filament associated protein found in smooth muscle as a potential modulator of contraction. Five mouse monoclonal antibodies (mAbs CP1, CP3, CP4, CP7, and CP8) were prepared against chicken gizzard alpha-calponin. The CP1 epitopic structure is conserved in smooth muscles across vertebrate phyla and is highly sensitive to CNBr cleavage in contrast with the chicken-specific CP4 and the avian-mammalian-specific CP8 epitopes that are resistant to CNBr fragmentation. Using this panel of mAbs against multiple epitopes, only alpha calponin was detected in adult chicken smooth muscles and throughout development of the gizzard. Western blotting showed that the calponin content varied among different smooth muscle tissues and correlated with that of h-caldesmon. In contrast with the constitutive expression of calponin in phasic smooth muscle of the digestive tract, very low levels of calponin were detected in adult avian tracheas and no calponin expression was detected in embryonic and young chick tracheas. These results provide information on the structural conservation of calponins and suggest a relationship between calponin expression and smooth muscle functional states. PMID- 9213428 TI - ATP- and EGF-stimulated phosphatidulinositol synthesis by two different pathways, phospholipase D and diacylglycerol kinase, in A-431 epidermoid carcinoma cells. AB - The [(3)H]inositol incorporation into the membrane fraction of A-431 human epidermoid carcinoma cells was markedly increased by stimulation of the cells with either epidermal growth factor (EGF), ATP, bradykinin, or a calcium ionophore A23187 in the presence of 1 mM extracellular calcium ions; most incorporated [(3)H]inositol was found to have accumulated as phosphatidylinositol (PI). The EGF- and ATP-stimulated PI synthesis was inhibited by two protein kinase C inhibitors, staurosporine and 1-(5-isoquinolinesulfonyl)-2 methylpiperazine dihydrochloride (H-7), and an intracellular calcium chelator, 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid acetoxymethyl ester (BAPTA/AM), but not by the calmodulin antagonist N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide hydrochloride (W-7). Pretreatment of cells with pertussis toxin (IAP, islet-activating protein) inhibited the PI synthesis, [Ca(2+)]i elevation, and inositol trisphosphate (IP(3)) production by ATP, suggesting that the phospholipase C(PLC) system coupled with IAP-sensitive G protein is involved in the ATP-stimulated PI synthesis. On the other hand, the ATP stimulation increased the release of [(3)H]choline and [(32)P)phosphatidic acid (PA) from radiolabeled cells, and such release was not inhibited by IAP. In the presence of n-butyl alcohol, which prevents the production of PA by generation of phosphatidylbutanol, the ATP-stimulated PI synthesis was reduced. Because n-butyl alcohol did not inhibit IP(3) production and [Ca(2+)]i elevation, this fact suggests that the lAP-insensitive PLD system is involved in the ATP-stimulated PI synthesis. In A-431 cells, the stimulation of P(2)-purinergic receptors appears to activate the IAP-sensitive PLC system and IAP-insensitive PLD system, both of which are essential for the stimulation of PI synthesis. The present results imply the general prospect that ligand stimulation, which mobilizes second messengers and consumes their precursors, simultaneously provokes the pathway to synthesize and salvage the second messenger precursors as well. PMID- 9213429 TI - Purification and metal ion requirements of a candidate matrix metalloproteinase: a 41 kDa gelatinase activity in the sea urchin embryo. AB - Using substrate gel zymography, the sea urchin embryo was found to express a dynamic pattern of gelatinase activities with a 41 kDa species persisting throughout the course of embryonic development. We have purified to near homogeneity the 41 kDa gelatinase in the sea urchin egg. In both qualitative and quantitative assays, the 41 kDa gelatinase activity was inhibited by ethylenediaminetetracetic acid but not the serine protease inhibitor, phenylmethylsulfonylfluoride, or the chelating agent, 1,10-phenanthroline. Activity could be restored to the inactivated gelatinase by each of several divalent cations: Ca(2+) > Mn(2+) > Mg(2+) > Cu(2+). Cadmium and Zn(2+) were largely ineffective at reconstituting the inactivated enzyme. In metal ion binding assays, the relative apparent affinities of the metal ions for binding to the gelatinase were determined to be Zn(2+) > or = Cd(2+) > or = Ca(2+) > Mn(2+) > Mg(2+) > Cu(2+). While the gelatinase is clearly a metalloproteinase, metal ion binding per se is not sufficient for activity. The 41 kDa gelatinase exhibited selective substrate utilization, being most active with gelatin, substantially less active with casein, and inactive towards bovine haemoglobin and bovine serum albumin as substrates. The substrate specificity and metal ion requirements suggest that this species is a member of the matrix metalloproteinase class of extracellular matrix remodelling enzymes. PMID- 9213430 TI - Modelling of purine nucleoside metabolism during mouse embryonic development: relative routes of adenosine, deoxyadenosine, and deoxyguanosine metabolism. AB - The individual activities for adenosine kinase, deoxyadenosine kinase, adenosine deaminase, deoxyguanosine kinase, and purine nucleoside phosphorylase were determined during days 7 to 13 of mouse embryonic development. Adenosine deaminase increased 74-fold between days 7 and 9; deoxyadenosine kinase increased 5.4-fold during the same interval. Adenosine kinase, deoxyguanosine kinase, and purine nucleoside phosphorylase exhibited less than 2-fold changes in activity between days 7 and 13. Using Michaelis constants for each enzyme and the maximal velocities determined from enzyme assay, the relative routes of adenosine and deoxyadenosine metabolism via phosphorylation or deamination were modeled as a function of nucleoside concentration for days 7 through 13. For days 7 and 8, phosphorylation of adenosine is the principle route of metabolism at physiological concentrations. A switch occurred at day 9 and following where deamination is at least 5-fold greater than phosphorylation at all substrate concentrations. Deoxyadenosine phosphorylation was at most 10% of deamination at day 7 and then declined to less than 1% for days 9 to 13. Phosphorolysis was the principle route of deoxyguanosine metabolism through the 7 to 13 day period. Thus catabolism rather than phosphorylation was the principle pathway for purine deoxynucleoside metabolism during this period. PMID- 9213431 TI - Purification of P26h: a hamster sperm protein. AB - P26h is a 26 kDa glycoprotein, located on the acrosome cap of hamster spermatozoa, involved in the species specificity of gamete interaction. We have purified this protein from hamster spermatozoa collected from the distal cauda region of the epididymis. Its purification was realized following a three-step procedure: detergent extraction, ion-exchange chromatography, and chromatofocusing. Protein partitioning using Triton X-114 (the first step) showed a ratio of 5:1 between the resulting aqueous and detergent phase. P26h was found almost exclusively in the aqueous phase where it represented about 10-12% of the total protein content. When the desalted aqueous phase was loaded on a carboxymethyl column for cation-exchange chromatography, about 80% of the proteins did not bind to the matrix and were eliminated. P26h was eluted from the column with a linear gradient of salt. At this point, P26h had a rate of purification estimated at 45-55%; three other major proteins of <21, 45, and 63 kDa remained in the sample. These undesired proteins were eliminated by submitting these samples to chromatofocusing using a PBE 94 polybuffer exchanger column. Indeed, P26h was collected almost in the dead volume of the column while the other proteins were eluted much later. Two-dimensional gel electrophoresis and Western blotting were performed to determine the purity of P26h. Only one major spot was detected, confirming the purity of P26h. Usefulness of this purified sperm antigen in the understanding of the physiology of mammalian fertilization is discussed. PMID- 9213432 TI - Isolation and partial characterization of a high molecular weight anionic glycoconjugate from transforming growth factor-beta treated bovine articular chondrocyte cultures. AB - A high molecular weight anionic glycoconjugate was isolated from the media of the transforming growth factor-beta treated chondrocyte cultures by anion-exchange chromatography on DEAE-Sephacel and Mono Q columns and was partially characterized. This high molecular weight anionic glycoconjugate was not detected in the non-treated (control) cultures. Characterization studies showed that the glycoconjugate is a non-reducible, non-collagenous glycoprotein containing O linked, N-linked, and sialic acid substituted carbohydrate units. The isolated glycoconjugate stained "blue" with Stains All and migrated as a single band on sodium dodecyl sulfate gradient gels (2.5-10% acrylamide - diallyl tartardiamide) at an estimated molecular weight of 540 000. Amino acid and amino sugar analyses showed that it is rich in aspartic acid--asparagine, glutamic acid--glutamine, alanine, proline, and glycine, and contains galactosamine and glucoasmine. This transforming growth factor-beta inducible glycoprotein may be involved in cell differentiation and in the cartilage repair process. It may also be used as a marker to localize the biological activity of transforming growth factor-beta in articular cartilage. PMID- 9213433 TI - Structural characterization of the O-antigenic polysaccharide of Escherichia coli serotype 017 lipopolysaccharide. AB - The structure of the O-polysaccharide component of the lipopolysaccharide produced by Escherichia coli 017 (ATCC 23512) was determined by the use of methylation, periodate oxidation, one- and two-dimensional nuclear magnetic resonance spectroscopy, and mass spectrometric methods. The O-polysaccharide was found to be a high molecular weight polymer of repeating branched pentasaccharide units composed of D-mannose, D-glucose, and 2-acetamido-2-deoxy-D-glucose residues (3:1:1) and had the structure (formula: see text). PMID- 9213434 TI - Ferriheme and ferroheme are isosteric inhibitors of fatty acid binding to rat liver fatty acid binding protein. AB - In addition to fatty acids, liver fatty acid binding protein (L-FABP) also interacts with ferriheme, which it binds with an affinity approximately one order of magnitude greater than that for oleic acid. We have, therefore, examined the effect of ferroheme and ferriheme on the binding of oleate to rat L-FABP also called heme-binding protein. Both oxidation states of heme behaved as isosteric inhibitors for the binding of the fatty acid confirming a common binding site. The reduced form of heme (Fe(II)) is a threefold better competitor of oleate binding than ferriheme. To show whether the diffusion of heme would be affected by the presence of the binding protein, we measured the effect of the fatty acid binding protein on the diffusional flux of a water-soluble heme derivative, iron deuteroporphyrin. The diffusional flux of iron-deuteroporphyrin did not change in the presence of the protein. This suggested that the binding affinity of fatty acid binding protein for iron-deuteroporphyrin is too great to allow rapid equilibrium between bound and unbound ligand across the system in an appropriate time frame. PMID- 9213435 TI - Dietary induction of pancreatic cholesterol esterase: a regulatory cycle for the intestinal absorption of cholesterol. AB - Atherosclerosis has a strong dietary basis without a proven molecular mechanism for cholesterol absorption. To investigate the potential role of pancreas in this process and its interaction with the two dietary forms of cholesterol (free and esterified), we undertook to study the role of pancreatic cholesterol esterase in cholesterol absorption. The results showed that (i) cholesterol esters contribute a disproportionately high fraction of absorbed dietary cholesterol, (ii) rates of intestinal cholesterol absorption are related to pancreatic cholesterol esterase activity, (iii) mRNA specific for pancreatic cholesterol esterase is induced 15 fold by dietary sterol esters and 10-fold by free sterol, (iv) the induction of cholesterol esterase mRNA is reversible, and (v) free cholesterol transport into cultured human intestinal cells is enhanced 300% by pancreatic cholesterol esterase. These data implicate pancreatic cholesterol esterase as pivotal in a metabolic loop under positive feedback control for the absorption of dietary cholesterol, whether free or esterified. PMID- 9213436 TI - Effects of aging and diet on the accumulation of dolichol in rat tissues. AB - A diet containing 15% (w/w) fat and 20% (w/w) of either casein or soy protein was fed to rats between 1 and 18 months of age. The effects of these dietary proteins on the accumulation of cholesterol and dolichols in kidney, spleen, brain, and heart were studied. The amount of cholesterol in these tissues was not influenced by the diet. In kidney and spleen, the amount of dolichols in rats fed the experimental diets was 50-70% higher than those in rats fed the lab chow diet. The contents of spleen dolichols in rats fed the soy protein diet tended to be higher than those in rats fed the casein diet. The amount of dolichols in heart and brain was not influenced by the diet. The proportion of spleen dolichyl fatty ester in rats fed the experimental diets was higher than that in rats fed the lab chow. The distribution of the dolichol isoprenologues was not influenced by the diet. There was a shift in the dolichol isoprenologues in kidney and spleen toward ones of lower chain length until 2 months of age, and after that there was no change. However, in heart and brain they shifted toward ones of lower chain length with aging. Our results suggested that dolichol metabolism may be influenced by fat content in the diets and differed among rat tissues. PMID- 9213437 TI - Glucose inhibits phenobarbital-induced delta-aminolevulinate synthase expression in normal but not in diabetic rat hepatocytes. AB - In the present work, we demonstrate the presence of a glucose inhibitory effect on the phenobarbital-mediated induction of the delta-aminolevulinate synthase mRNA in normal rat hepatocytes, consistent with the results obtained with the delta-aminolevulinate synthase activity previously reported. This "glucose effect" can be prevented by adding cAMP, adenylate cyclase activators, or a phosphodiesterase inhibitor. Delta-Aminolevulinate synthase mRNA half-life is not modified in the presence of phenobarbital or glucose. When the same experiments are performed using diabetic cells, no glucose effect is observed, even when the endogenous cAMP content is lowered to normal levels. The results obtained in this study suggest that glucose decreases delta-aminolevulinate synthase biosynthesis by acting at a pretranslational step. Assuming that the glucose effect operates by a repression mechanism exerted by metabolites derived from or related to glucose, the present results may reflect a derangement in the formation of these metabolites as a result of the abnormal metabolism operating in the diabetic state. PMID- 9213438 TI - Accumulation of natural and synthetic betaines by a mammalian renal cell line. AB - Intracellular accumulation of different betaines was compared in osmotically stressed Madin Darby canine kidney (MDCK) cells to model the betaine accumulation specificity of the mammalian inner medulla and to show how this accumulation differed from that of bacteria. All betaines accumulated less than glycine betaine. Arsenobetaine (the arsenic analogue of glycine betaine) accumulated to 12% of the glycine betaine levels and the sulphur analogue dimethylthetin accumulated to >80%. Most substituted glycine betaine analogues accumulated to 2 5% of intracellular glycine betaine concentrations, however, serine betaine accumulated to <0.5% of glycine betaine levels. Inhibition studies to distinguish the betaine ports were performed by the addition of proline. Butyrobetaine and carnitine accumulation was not proline sensitive, whereas that of other betaines was. As with glycine betaine, the accumulation of propionobetaine and dimethylthetin was proline sensitive and osmoregulated. Pyridinium betaine was accumulated by both proline-sensitive and -insensitive systems, with a small increase under osmotic stress. High concentrations (10 times that of glycine betaine) of the dietary betaines proline betaine and trigonelline inhibited total betaine accumulation. Because alpha-substituted betaines are accumulated by bacteria and not by MDCK cells, these betaines may be the basis for design of antimicrobial agents. PMID- 9213439 TI - Mitogenic stimulation of primary cultures of lung epithelial cells by linoleic acid. AB - The role of linoleic acid (18:2 n-6) in stimulating proliferation of normal lung epithelial cells in vitro is investigated. When 18:2 n-6 is present with insulin (I) and cholera toxin (CT), growth is stimulated synergistically. In the presence of indomethacin (10 mu M), an inhibition of proliferation is observed in I,CT, and 18:2 n-6, which can be reversed by the addition of exogenous prostaglandin E(2) (PGE(2)). Incorporation of [(14)C]18:2 n-6 with lipid-independent I, CT, and cortisol and lipid-dependent I, CT, and 18:2 n-6 conditions suggests differences in mobilization of 18:2 n-6 from the phospholipid (PL) fractions between 2 and 8 days. The decline of [(14)C]18:2 n-6 in PL fractions with lipid-dependent condition suggests that free 18:2 n-6 may be available for metabolism by the cyclooxygenase pathway. In non-proliferative cultures, an accumulation of the label in the PL fraction is observed. Proliferation in lipid-dependent conditions appears to be due to the mobilization of 18:2 n-6 whereas proliferation in lipid independent conditions appears to be independently controlled. PMID- 9213440 TI - The activation of beta-galactosidase (E. coli) by Mg(2+) at lower pH values. AB - The activation of beta-galactosidase (E. coli) by Mg(2+) at pH values below 7.6 was studied. If the Mg(2+) concentration was high enough, the k(cat) values at pH values down to 5.0 remained at the same high level as at pH 7 and 7.6 (600-620 s (1) with o-nitrophenyl-beta-D-galactopyranoside as the substrate). Very high concentrations of Mg(2+) (greater than 100 mM at pH 5) were, however, needed to saturate the Mg(2+) site at lower pH values. The Km values at low levels of Mg(2+) were high at every pH but they decreased and approached the same low value at every pH (about 0.13 mM) as the [Mg(2+)] was increased. These data indicate that it is difficult to bind Mg(2+) at lower pH values, but the k(cat) and K(m) values of the enzyme, and therefore the rates of galactosylation (k(2)), degalactosylation (k(3)), and binding (K(s)), do not change substantially as a function of pH provided that a Mg(2+) is bound to the enzyme. The data also showed that Mg(2+) and protons compete for the same site. Analysis by plotting log [Mg(2+)](mid) vs. pH showed that the binding of Mg(2+) to the free enzyme involves two groups with pK(a) values in the vicinity of 7 and one group with a pK(a) value near 5.5. (The values referred to as [Mg(2+)](mid) are the Mg(2+) concentrations that resulted in k(cat) values midway between basal and maximum.) The "apparent" pK(a) values of the groups when a Mg(2+) was bound (at saturating [Mg(2+)]) all appeared to be below 5.0. PMID- 9213441 TI - Altered distribution of the promyelocytic leukemia-associated protein is associated with cellular senescence. AB - The disruption of the normal function and nuclear localization of the promyelocytic leukemia-associated protein (PML) may play a major role in the pathogenesis of acute promyelocytic leukemia. PML, which is concentrated in nuclear bodies (PML bodies), has been shown to have growth- and transformation suppressive properties. In this study, we have examined the intranuclear distribution of PML in a conditionally immortalized human cell line (IDH4) in which both proliferation and immortalization are dependent on the presence of SV40-encoded large T-antigen (SV40T). Expression of SV40T is controlled by a dexamethasone (Dex)-inducible promotor. Suppression of SV40DT (Dex removal) in IDH4 cells causes G1 arrest and expression of the senescent phenotype. This is accompanied by a redistribution of PML in most cells from the usual pattern containing only spherical bodies to a pattern, containing large doughnut-like or fiber-like structures in addition to the spherical bodies. This change in pattern is reversed when phenotypically senescent IDH4 cells are stimulated to proliferate again by SV40T-induction. Moreover, we find that there is a similar change in the PML pattern between young and senescent or serum-starved young IMR90 human fibroblasts, from which IDH4 cells are derived. However, fewer serum starved cells contain large PML bodies than senescent cells. Our observations suggest senescence, although it may be partly related to growth arrest. Using three-dimensional fluorescence digital imaging microscopy, we have found that the apparently doughnut-like PML structures have a cylindrical or egg-shaped form and that PML is concentrated to the outer shell of the structure. PMID- 9213442 TI - [Endotoxemia in experimental acholia]. PMID- 9213444 TI - [Mechanism of potentiation of duodenum contraction after stimulation of the sympathetic fibers]. PMID- 9213443 TI - [Is chronic serotonin deficiency the basis of diabetic and age-related angiopathy?]. PMID- 9213445 TI - [Determination of three forms of plasma kallikrein and its adsorption on kaolin using the colorimetric method]. PMID- 9213446 TI - [Effect of specific antibodies on morphological characteristics of murine oocytes]. PMID- 9213447 TI - [Changes in the physico-chemical and functional properties of human hemoglobin after laser irradiation in vitro]. PMID- 9213448 TI - [Effects of prolactin on DNA biosynthesis, cholesterol content, and steroidogenesis in the adrenal cortex of guinea pigs treated with dexamethasone]. PMID- 9213449 TI - [Generation of oxygen free radicals and clastogenesis in bone marrow cells of MRL/1 mice]. PMID- 9213450 TI - [Factors influencing the structural-functional properties of erythrocyte membranes in pregnant women]. PMID- 9213451 TI - [The effect of liposomal form of L-Dopa on the development of parkinsonian syndrome in mice]. PMID- 9213452 TI - [Reduced blood trauma in vitro in the electromagnetic field]. PMID- 9213453 TI - [Effect of L-lysine-alpha-oxidase on the synthesis of cell proteins, targets of autoimmune pathology in HIV infection]. PMID- 9213454 TI - [The role of thiols in the stimulation of soluble guanylate cyclase by a new class of enzyme activators generating nitric oxide]. PMID- 9213455 TI - [Characteristics of morphological differentiation of hepatocytes in ontogenesis of different animals depending on the type of nutrition]. PMID- 9213456 TI - [The effect of leu-enkephalin analog dalargin on DNA synthesis in the myocardium and tongue epithelium of albino rats at early stages of postnatal ontogenesis]. PMID- 9213457 TI - [The effect of gamma-irradiation on the level of transferrin in the plasma of mice and the degree of its glycosylation]. PMID- 9213458 TI - [Changes in the nonspecific and immunological resistance during acute dichlorethane poisoning]. PMID- 9213459 TI - [Early stages in the mechanism of action of glucocorticoids on human platelets. The effect of hydrocortisone on platelet aggregation]. PMID- 9213460 TI - [Effect of aspartic acid derivatives, N-acetyl-aspartate and its phosphonic analog PIR-87-6-0, on dopamine release from the rat striatum during perfusion in vitro]. PMID- 9213462 TI - [The effect of vitamin D on the humoral and cellular immune response and functional activity of peritoneal macrophages]. PMID- 9213461 TI - [Formation of elevated sensitivity to seizures induced by a single administration of norbornan]. PMID- 9213463 TI - [Suppressor effect of immature erythroid cells on B-cell proliferation]. PMID- 9213464 TI - [Immunotropic effect of non-laser monochromatic irradiation]. PMID- 9213465 TI - [Dynamics of sister chromatid exchanges during repeated administration of thiophosphamide in vivo]. PMID- 9213466 TI - [Repopulation defect of hematopoietic stem cells after retrovirus transfer of foreign gene]. PMID- 9213467 TI - [Correlation between the molecular heterogeneity of ceruloplasmin mRNA and multiple molecular forms of ceruloplasmin synthesized in the rat liver]. PMID- 9213469 TI - [The effect of topoisomerase II inhibitor, vepesid, on binding of doxorubicin with DNA in tumor cells sensitive to anthracycline]. PMID- 9213468 TI - [Prostaglandins E in the primary tumor, metastases, and ascitic fluid of patients with ovarian cancer]. PMID- 9213470 TI - [Isolation of pectin from Amaranthus cruentus and study of its effect on the function of isolated rat heart]. PMID- 9213471 TI - [Localization of collagen-synthesizing cells in normal and atherosclerotic intima of human aorta]. PMID- 9213473 TI - Bibliography. Current world literature. Chromosomes and expression mechanisms. PMID- 9213472 TI - [Clinical significance of tumor markers CYFRA 21-1 and neuron-specific enolase in lung cancer]. PMID- 9213474 TI - [Health status of the population as a criterion in the assessment of the quality of life]. AB - The paper analyzes the present health status of the population, including mortality of able-bodied individuals, examines the changes in age-specific mortality rates. This also gives the results of studies into the causes of increased deaths from various unfavourable exposures to occupational, environmental, socioeconomic and other factors. The paper also presents epidemiological cohort surveys of workers exposed to lead and outlines the objectives and tasks of the section "Environmental Epidemiology" of the Environmental Control Project. PMID- 9213475 TI - [The Nation's health as a social mechanism of society's stable well-being]. AB - The paper provides evidence for the need of creating new social mechanisms of nation's health promotion as a factor that influences the stability and well being of society. To accomplish these tasks, health should be regarded as a polysystem concept that includes 1) a developing human being as a source of nation's intellectual resources; 2) economy as a tool for social welfare; 3) ecology as a predictor of the influences of working conditions; and 4) social policy as a a mechanism of controlling the priorities of prevention and treatment. New principles of new management organizational forms for health protection have been developed to implement the stated theoretical points. PMID- 9213476 TI - [Systems evaluation of physiological functions of humans at a workplace]. AB - The paper gives experimental findings of the application of the Anokhin theory of functional systems to the evaluation of human physiological functions in a working place. It is shown that the correlations of the heart and respiration rates with the effective productive activity quanta may characterize the specific features of autonomic regulation in workers and objectively suggest the occurrence of their psychoemotional stress in their working place. Disintegrated autonomic correlations in the productive activity quanta reflects impaired multicomponent interactions of homeostatic functional parameters, by ultimately causing an abnormality. PMID- 9213477 TI - [Psychophysiological reserves of occupational health of man]. AB - The paper outlines experimental studies to assess the occupational health of individuals engaged in hazardous trades. The quantitative parameters reflecting the relationship between the health status and the decreased action reliability are defined. Psychophysiological correlations between the diminished action reliability and age are ascertained. Mechanisms of psychophysiological readiness for emergencies are assessed. A theoretical concept of new developments for enhancing the skill of the personnel engaged in hazardous trades by forming the body's life-time limiting functions ensuring high compensation is accounted. A psychological strategy of consciousness to use informational energy fields in the interests of reliability enhancement in emergencies is worded. PMID- 9213478 TI - [Conceptual approaches to the diagnosis of stress-induced functional disorders in humans under conditions of industrial activity]. AB - In accordance with the fundamental concept of the systems symmetrical approach, a model of the body's "ideal functional status" and a polyparametric method for diagnosing the human functional status (the method has been patented in Russia) were developed, on the basis of the unified set of amplitude and time parameters of a complex of physiological parameters (ECG, etc.). The systems symmetrical approach is basically new in defining the ratio of harmonic parameters acting as new diagnostic evidence for the health status. The unchanging ratio of parameters of autovariation processes in the cardiac, vascular, and respiratory systems, their changes in the development of the adaptative syndrome and the value were examined in healthy individuals at rest and during exercise an intensive productive activity (point brazing at a plant of electron devices), and in the trainees. It was demonstrated that the mechanism of development of adaptative processes was associated with the synchronization of autovariations in the cardiorespiratory and vascular systems and the involution of adaptative processes, their break, depletion, i.e. stress, were determined by the imbalance of relationships and the desynchronization of autovariations in these systems. To define a set of cardiorespiratory and hemodynamic parameters is necessary, but insufficient to evaluate (diagnose) the health status and health-disease intermediate diseases (stress and adaptative overtension). The ratio harmonicity of these parameters, i.e. well-balanced relations between the subsystems is the main characteristic. The assessment of the human health status in terms of the reference-the model of the ideal status as the ideal health status without using the complex and indefinite concept of the normal and moderate health status-is conceptual for polyparametric status estimation. PMID- 9213479 TI - [Problems of occupational medicine in Russia: models of present-day practices and strategy]. PMID- 9213480 TI - [Medical and social aspects of the protection of working women's health]. AB - The paper presents statistic data on the Russian females engaged in national economy and its individual industries, results of studies into the reproductive health of working women (their health during pregnancy and nonpregnancy, gynecological morbidity). It also gives data on the relationship of reproductive health with the factors of production. PMID- 9213481 TI - [Protection of occupational health as an ergonomic problem]. AB - The paper outlines the scientific and applied aspects of ergonomic improvement of the protection and salvation facilities for aircraft specialists under the present environmental conditions of their operation. A concept of ergonomic enhancement of the efficiency of these facilities under the combined influence of unfavourable factors is substantiated and forwarded, which envisages the possibility of improving professional skills and the preservation of the working capacity and occupational health of aircraft specialists by making a fuller account when the facilities are designed and the characteristics of an operator are examined. An analysis has been made of the methods which may be useful in evaluating the influence of environmental factors on the quality and tension of flying activities and in assessing the impact of introduction of ergonomic recommendations on the optimization of protective means against them. PMID- 9213482 TI - [Methodological and organizational aspects of psychophysiologic care in the military training of aviation specialists]. PMID- 9213483 TI - [Psychophysiologic pattern of pilot's activity during flight as a basis for setting up his workplace]. AB - The psychophysiological pattern of a pilot is considered on the basis of experiments made during flights. Five components of his activity, which may be estimated, are identified. These include sensory, motor, autonomic, occupational, and psychological components. The "reserves of attention" are singled out as an objective independent parameter that characterizes the activity of a pilot. Five determinants of the level of physiological responses are identified in a pilot during flight. Their mechanisms are most efficiently analyzed in terms of the basic concepts developed by P.K. Anokhin, P.V. Simonov, K.V. Sudakov. The objective activity estimates make it possible to assess the adequacy of a working place to the problems to be solved, to apply the anthrocentric principle in working out the pilot's working place by taking into account his psychophysiological capacities. PMID- 9213484 TI - [Evaluation of technogenic radiation exposures from ecological and physiological points of view]. AB - Many years' experience with complex supervision of the environmental radiation situation, the specific features of the population's physiological and health status after radiation accidents (the Chernobyl, Kyshtym and other accidents). The paper emphasized the necessity of having a knowledge of the specific features of the influence of a radiation factor (and its components) on establishing the radiation dose for the population due to the environmental peculiarities of the radiation background in different regions and abnormal zones of the Earth. The paper provides evidence for the need to apply an environmental and physiological approach to working out the standards of radiation doses for the inhabitants of post-accident polluted areas. To this end, a concept of the regional standard of exposure is proposed for different regions of the Earth with regard to the specific natural radiation background, technogenic and medical "additives" on making diagnostic and therapeutical efforts by using X-rays and radiation therapy. A new concept is offered to develop the life-support systems in the areas exposed to radiation contamination, whose main goal is to minimize radiation exposures with regard of data on each area-specific data on the regional standard of the pre-accident radiation background, technogenic background, medical additives and prediction of future radiation doses. Exposure of the population to radiation is minimized by reasonably operating its components and making special life regimens in the contaminated areas. PMID- 9213485 TI - [Use of electrical impedance tomography in detection of latent diseases during mass screening]. AB - The paper deals with the use of the present-day highly effective method of introscopy electric impedance tomography (EIT) for diagnosis latent abnormalities during mass screening. EIT images viscera and tissues on the basis of their electric conductivity differences. Reconstruction of the images reflecting the distribution of electric conductivity values in the body is made from the recording potentials onto the body's surface while passing currents through the electrodes located along the circumference of the body's part under study. The 16 electrode EIT system developed by Dipol (Moscow) was used to examine the distribution of electric conductivity values in the extremities and chest organs in healthy volunteers of both sexes and in patients with various pulmonary diseases. PMID- 9213486 TI - [Rehabilitation of patients with ischemic heart disease after aortocoronary bypass surgery]. AB - The paper outlines some experience in rehabilitating 1250 patients with coronary heart disease (CHD) undergone aortocoronary bypass surgery (ACBS). Rehabilitative measures promoted more rapid and maximally full recovery of physical fitness and cardiorespiratory parameters. The recommended groups of physical activity allow one to differentially approach to the choice of the optimum training regimen and hence to achieve the maximum training effect. The groundlessly high proportion of disabled patients with CHD after ACBS is due not to the quality of surgical treatment, but to the absence of a rehabilitative programme for this group of patients and to the presence of obsolete standard acts for definitions of disability groups by the medical-and-labor examination commission. PMID- 9213488 TI - [Health insurance in the Russian Federation]. AB - The need to reform health care in Russia became evident in the late 1980s when due to socioeconomic crisis, the government could not cover the expenses connected with this field and put the up-to-date expensive technologies into life. The introduction of the compulsory health insurance system (CHIS) is aimed at: 1) obtaining an additional financial source for health care by making the goal-oriented stable rates of deductions from the wage fund; 2) protecting the Russian Federation citizens' rights to have free medical aid of the guaranteed scope; 3) enhancing the quality of medial care delivered to the population by introducing a mechanism of movement of funds paid for a patient; 4) paying for medical care in relation to the volume and quality of the work done by simultaneously controlling the stipulated use of funds. Three-year experience of CHIS in Russia has indicated that there is a real mechanism of reformation and government regulation of health care under the conditions of transition to the market, with the interests of the general population and medical personnel in mind. Obvious legal, organizational, technological, and psychological problems and disadvantages have been found at all management levels, which are an obstacle in the way of the reforms and which whip up social tension and call for prompt decisions. PMID- 9213487 TI - [Biomedical aspects of health preservation strategy: role of catecholaminergic neuronal populations]. AB - Lifestyle, environmental factors, genetics, and medical care are the main factors that determine the health status of man. Of particular attention are biological mechanisms ensuring the body's adaptation to constantly changing environmental conditions. The noradrenergic neuronal populations, the sympathetic nervous system in particular, modulate metabolic processes and supports a variety of activities, making them relevant to changing living conditions. There is a clear correlation between the life span and the number of sympathetic nerve cells functioning during postnatal ontogenesis. The exposures that reduce the activity of peripheral and central noradrenergic neurons and slow down aging processes in them loosen the relationships between the inner and outer world to prevent hyperactivity and to prolong life. PMID- 9213489 TI - [Nitric oxide (NO) increase sensitivity and decreases desensitization of the neuron to the stimulating effect of glutamate]. PMID- 9213490 TI - [Ability of cat visual system neurons to assess the spatial brightness gradient]. PMID- 9213491 TI - [Effectiveness of a substance protecting against aging depends on the viability of an experimental population. Study of the life expectancy of D. melanogaster]. PMID- 9213492 TI - [Directed transfer of the human apolipoprotein A-I gene and bacterial genes markers into rat liver]. PMID- 9213493 TI - [Prognostic factors in organ-confined renal epithelial tumors]. AB - Recent application of molecular cytogenetic techniques to the evaluation of epithelial renal cell tumors have showed some characteristic combinations of genetic alterations within the chromosomal DNA. Moreover each group of abnormalities has been correlated with peculiar tumor morphology. The new classification of renal cell neoplasms proposed by G. Kovacs, based on specific genetic alterations, and histologic pattern, together with the morphologic and pathologic features that correlate with survival has been discussed. PMID- 9213494 TI - [Multifocal renal carcinoma: anatomic-clinical aspects]. AB - Between 1992 and 1996 we examined 387 kidneys after nephrectomy for renal cell carcinoma. The capsules were removed and the kidneys were serially sectioned at 5 mm intervals; cortical and intraparenchymal nodules were examined histologically. Of the 387 kidneys 12 (3%) contained multifocal renal cell carcinoma and only 3 with clinically overt tumor ut to 3 cm. in diameter. We conclude that partial nephrectomy should be widely accepted in patients with small renal cell carcinoma in presence of a normal controlateral kidney. PMID- 9213495 TI - [Conservative therapy of renal parenchymal neoplasia]. AB - We review our overall experience in 1375 patients, who underwent surgery for renal cell carcinoma in the Departments of Urology of Brescia and Bergamo from 1983 to 1996. 185 (13.4%) patients had nephron-sparing surgery: imperative procedure was performed in 74 cases, while an elective surgery was done in 111 patients. Three years minimal follow up was considered in order to evaluate the outcome of surgical treatment in 48 patients who underwent imperative nephron sparing surgery and in 73 with an elective procedure. Disease specific survival was 80.8% in the first group and it was 97% in the latter. PMID- 9213496 TI - [Imaging of small renal tumors]. AB - The advent of ultrasound and computed tomography resulted in a great increase in detection and diagnosis of small renal parenchymal tumors. These are mainly slow growing tumors, without metastatic disease and with possible multicentricity at the diagnosis. Moreover there is not agreement about the best treatment for the small (< 3 cm) renal cell carcinoma. In this paper the role of ultrasound, computed tomography and magnetic resonance for detection and characterization of the small renal masses is discussed. On occasion it is possible to obtain the tissue characterization of a solid renal mass by diagnostic imaging (for example angiomyolipomas); however, most frequently, solid renal masses have an aspecific appearance. The majority of problematic renal masses have cystic components. The diagnosis of simple cyst is based on few simple but rigid criteria: homogeneous water density, very thin wall, well defined and sharp interface with renal parenchyma, lack of contrast enhancement. When there are intracystic septae, thickened wall or increased density, the cyst is "complicated". In these cases the classification suggested by Bosniack can be helpful. Bosniack class-1-lesions are simple cysts; they do not require any further work-up. Bosniack class-2 lesions are minimally complicated but reliably benign cysts (thin wall, thin calcifications, thin septae). Some of these lesions require follow-up; and the majority of them do not. Class-3-lesions have thick septae, thick calcifications and thick and irregular walls, but not contrast enhancement. In most cases these lesions require surgical exploration for diagnosis and therapy. Bosniack class-4 lesions are clearly malignant; they are indicated by contrast enhancing regions within cysts. They always require surgery. PMID- 9213497 TI - [Conservative treatment of renal cell carcinoma: framework, incidence and classification]. AB - Recent interest in nephron sparing surgery for renal cell carcinoma has been stimulated by advances in diagnostic imaging, following an increasing number of incidentally discovered low stage renal cell carcinoma and good long term survival in patients undergoing this form of treatment. Tosaka et al reported a 5 years survival of 94.7% in patients with incidental renal cell carcinoma compared with 60.9% in diagnosed because symptomatic. Along with a diagnosis of carcinoma more and more premature, a whole string of little lesions is present, not easily identifiable by the recent diagnostic imaging. Tosaka and others examined renal lesions going by the ultrasonography as a check-up or as a first frame in patients suffering from microscopic hematuria; they proved that neoplastic lesions represent 5.4% of all the masses identifiable by diagnostic imaging. The frequent discovery of limited carcinoma, the difficulty in the diagnostic attribution and demonstration of the good survival of patients who were treated by a nephron sparing surgery, added to the one of patients undergone to radical nephrectomy, caused an interest in nephron-sparing surgery for incidental renal carcinoma also for patients with normal controlateral kidney and not very extended tumors, usually in peripheral sites. At the moment record of cases concerning nephron sparing surgery is quite limited, any way it shows a survival equal to 90% with only two local recurrences, reported only in one experience and caused by an incomplete resection or by multicentric neoplastic lesions. PMID- 9213498 TI - [Conservative surgery of parenchymal renal carcinoma: urologic data from Lombardy]. AB - We report the results of a questionnaire sent to various Urology departments in Lombardy about "Conservative Surgery in Renal Cell Carcinoma" which was the subject of discussion at the 50th meeting of the Lombardian Society of Urologist. 23 centres out of 34 i.e. 68% answered. 100% of the departments performed nephron sparing surgery, 91% in cases of imperative and elective indication and 9% only in imperative indication. 100% of the patients underwent preoperative staging with ultrasound and computer tomography. 48% treated in elective surgery only incidentally asintomatic discovered tumours, but 52% treated both incidentally and sintomatic ones. Elective surgery is suggested when the tumor has a diameter less than or up to 3 cm., from 3 to 5 cm., and more than 5 cm. in 48%, 48% and 4% of the urological departments respectively. 70% consider the importance of tumor location and do not perform partial nephrectomy when the tumor is intraparenchymal or in contact with the secretory tract. From a technical point of view 82% carried out partial nephrectomies while 18% carried out enucleation; 57% performed routinally frozen section on tumor bed and 61% do not performed lymphadenectomy. 83% believe in the multifocality problem. A macroscopic and microscopic haematuria does not condition the elective indication in 62% of the urology departments. The follow-up is carried out with ultrasonography alternated with CT in 78% of the departments and is continued for over 5 years in a likewise 52%. From 1990 to 1995, 3332 patients were surgically treated for renal cell carcinoma in 23 urology departments; 487 (14.6%) underwent nephron sparing surgery; 320 elective and 167 imperative indication. The local global relapse was 2.9% (14/487); 5.3% (9/167) in imperative group and 1.4% (5/320) in elective group. Conservative surgery in Lombardy will always try to have the golden standard treatment in the incidental, single renal cell carcinoma. PMID- 9213499 TI - [Conservative therapy in renal carcinoma: follow-up]. AB - Due to the increasing use of sophisticated imaging techniques, up to 30% of diagnosed renal cell carcinoma (RCC) are asymptomatic and diagnosed incidentally. Getting the cue from our personal survey of conservative renal surgery for renal cell carcinoma with a cancer specific survival of 95.5% after a mean follow up of 32.7 months, a review of the literature is illustrated: numerous studies have documented the technical success rate with this approach as well as long term disease free survival, comparable to that obtained by radical nephrectomy, in patients with unilateral, small, low stage tumors and normal opposite kidney. Patient selection is of extreme importance in case of partial resection in the presence of a normal contralateral kidney. The tumor must be < 3-4 cm, solitary, well delineated on CT, without invasion of the perinephric far or pyelocaliceal system (T1 and T2), easily resectable with at least 1 cm of healthy parenchyma. Only well informed patients who agree on a careful follow up after surgery can be candidates for kidney sparing surgery. In case of imperative surgery the follow up must be strict and personified for every single patient. Those patients who underwent a partial nephrectomy in presence of a normal contralateral kidney should be monitored with a conventional follow up monitored in order to detect an eventual local recurrence: 12 monthly ultrasonography and contrast enhanced CT scan alternately every 6 months for the first five years after surgery and then lifelong once a year by echography and/or CT scan. PMID- 9213500 TI - Sexually transmitted diseases quarterly report: syphilis in England and Wales. PMID- 9213501 TI - Testing for differences with the nonparametric Mann-Whitney U test. PMID- 9213502 TI - Intracranial bacterial infections in patients with AIDS. AB - Nontuberculous, nonsyphilitic intracranial bacterial infections in HIV-positive individuals may be nonspecific presentations of unusual organisms, such as R. equi, B. henselae, Nocardia sp. or L. monocytogenes. In addition, more common organisms, such as Staphylococcus, Streptococcus, or Salmonella sp. may cause an unusually severe infection. In general, the imaging appearance of these bacterial infections is similar to that in immunocompetent individuals. PMID- 9213503 TI - [Ecological aggression and adaptation mechanisms]. AB - The article presents the main ecological hazards and human adaptational mechanisms. Special attention is paid to chemicals being highly prevalent in environment and to mechanisms of their activation and deactivation by means of enzymes dependent on cytochrome P450. PMID- 9213504 TI - [Hygienic and immuno-allergologic aspects of the effects of formaldehyde and wood dust in furniture production]. AB - The study evaluated effects of threshold sensitizing levels of formaldehyde (and its combination with woody dust) on health of furniture production workers. The results enable to conclude that MAC of formaldehyde (or combined with woody dust) increases danger of symptomatic allergies and considerably alters immune reactivity, causing atopic immune state. Formaldehyde promotes general nonspecific diseases by inducing the hyperergic reactivity that exhausts immune defense mechanisms. PMID- 9213505 TI - [Role of A.S. Arkhipov in the development of occupational hygiene in Russia]. PMID- 9213506 TI - [Formulating problems of ecological diseases. Approaches to etiopathogenesis, classification and diagnosis of ecological diseases due to chemicals]. AB - Our long-standing clinical trials proved nonspecific and stress factors contributing into environmental diseases. The studies helped to define traumatic variant of biologic stress, to describe its trigger mechanisms. The authors analyzed physiologic mechanisms of stress-related diseases, role of autonomous nervous system in formation of environmental diseases. Clinical classification of acute and chronic environmental diseases is given, with consideration of their diagnosis. PMID- 9213507 TI - [Measurement of lead and cadmium in urine and blood by the inversion volta amperometry using SVA-IBM analyzer]. AB - The authors suggest a method evaluating lead and cadmium in human urine by means of new Russian device SVA-IBM (CBA-IBM). Inversion volta amperometry forms a basis of the method that is quite sensitive, precise and selective. Substances normal for mineralized urine and blood do not interfere with the evaluation. The method is recommended for evaluation of lead and cadmium for clinical, hygienic, toxicological purposes. Unlike polarographic method, the technique requires no metallic mercury and highly pure inert gas. PMID- 9213508 TI - [Allergic dermatoses caused by paint and varnish materials in water transport workers]. AB - The article deals with occurrence and course of allergic dermatoses among water transport workers in specific occupational and living conditions. The authors present data on structure of allergic dermatoses, features of their prevalence in connection with sex, length of service, occupational peculiarities. The results of immunologic, laboratory tests and instrumentation are also demonstrated. The study helped to elaborate therapeutic and prophylactic measures that induced the certain medical, social and economic effects. PMID- 9213509 TI - [Renal function in thallium intoxication]. PMID- 9213511 TI - [Acid resistance of erythrocytes in workers of the Astrakhan gas processing plant]. PMID- 9213510 TI - [Hygienic evaluation of work conditions in the combined production of fodder using Biotrin protein additives]. PMID- 9213512 TI - [Clinical aspects of stomach and duodenal ulcer in workers exposed to phenol and formaldehyde]. PMID- 9213513 TI - [Anion-produced surface active agents, derivatives of alkylbenzyl sulfonic acid: toxicity and hazards]. PMID- 9213514 TI - [Experimental data on substantiation of MAC of methyl dioxide in the workplace air]. PMID- 9213515 TI - [Regional aspects of health status of the population with regard to chemical pollution of the environment]. AB - The article summarises data on regional aspects associated with health state of population in connection with environmental pollution. Changes in health state of population inhabiting polluted territories are assessed through wide variety of parameters, in accordance with special Criteria. Notions are that no adequate investigations cover influence of environmental factors on infants and children mortality, only few works deal with evaluation of toxic chemicals in human biologic material. PMID- 9213516 TI - [Diagnostic signs of technogenic effects of the environment in inhabitants of Bashkortostan industrial cities]. AB - The article deals with examples associated with use of oncologic morbidity and mortality in evaluation of technogenic effects in residents of cities with developed petrochemical and oil-processing industries. Oncologic morbidity proved to characterize the public health adequately to ranking the territories according to their ecologic jeopardy. PMID- 9213517 TI - [Rhabdomyosarcoma of the orbit]. AB - The authors evaluated 9 years' experience with the diagnosis and treatment of embryonic rhabdomyosarcoma of the orbit in children. They evaluated in detail a group of 5 children treated and followed up for 1-9 years. Due to comprehensive surgical, radiation and chemotherapeutic treatment all patients survive and in 80% the visual function is preserved. For differential diagnostic problems, with regard to the variable manifestation of rhabdomyosarcoma which may imitate orbitocellulitis, chalaseon, epibulbar lipodermoid or papilloma, the authors emphasize the importance of rapid primary diagnosis by NMR and biopsy. In the treatment they appreciate greatly a combination of radiotherapy and chemotherapy which is a modern trend and can eradicate the tumour without radical surgery. PMID- 9213518 TI - [Prescription of special optical aids for visual defects. I]. AB - In 1993-1995 the authors prescribed in the Institute for the Treatment of Ophthalmological Defects and at the Second Ophthalmological Clinic in Prague special optic aids for 795 patients. The patients were divided by age into three groups. In the group under 18 years the main cause of the defect was atrophy of optic nerve, in the group between 18 and 60 years tapetoretinal degeneration and in the group over 60 year senile macular degeneration. Most frequently a magnifying glass was prescribed (36.16%), a prismatic monocular in 31.30%, a hyperocular in 27.46% and telescopic spectacles in 5.08%. PMID- 9213519 TI - [Prescription of special optical aids for visual defects. II]. AB - In 1993-1995 the authors prescribed in the Institute for the Treatment of Ophthalmological Defects and the Second Ophthalmological Clinic optic aids for 795 patients with impaired eyesight and evaluated their functional effectiveness. Optic aids for far distance were prescribed to 325 patients. Their mean natural vision (or with common correction) was 4.5/60. With the optic aid the mean vision improved to 6/15-12. Aids for short distance were prescribed to 758 patients. Their mean natural vision or with common correction was J. No 15; with the use of the prescribed aid it improved to J. No 5. PMID- 9213520 TI - [Occurrence of refractive errors in relation to retinopathy of prematurity]. AB - Results of the development of refractive errors in the group of prematurely born children with reversible ROP changes and those where cryoretinopexy due to ROP were done are presented. Ametropia was found higher (statistically significant with P = 0.05) in all the groups with prematurity comparing to the population of in-therm born children. The same result with regard to strabismus was found. Prematurity presumably leads to ophthalmic pathology. PMID- 9213522 TI - [Intubation of afferent lacrimal ducts--a review of techniques]. AB - The authors present a review of various techniques of long-term intubation of efferent lacrimal pathways by a silicone fibre. They use an original way of intubation and intubate the insertion probes of the intubation set from the lower nasal outlet by means of a titanium loop under direct optic control by means of an endoscope. The use of the endoscope makes it possible to reduce the number of complications and iatrogenic damage of the mucosa and it enhances the effectiveness of the procedure. The authors prefer the use of special intubations sets; for young children intubation sets made from soft metal are preferable. At present all their demands are met by the intubation set they developed in collaboration with the Technical University, Prague. During the period between April 1994 and August 1996 the authors used the described technique for bicanalicular intubation of the efferent lacrimal pathways in 127 children and in adult patients 6x. In 86 instances they used an intubation set of Geuder Co., 44 times their own intubation set. PMID- 9213521 TI - [Masquerading syndromes]. AB - Masquerade syndromes comprise a number of diseases imitating as to their clinical picture inflammation of the eye. These diseases comprise also malignant tumours. The authors present three patients where intraocular tumours (lymphoma, leukaemia, melanoma of the choroid) reminded of chronic uveitis. PMID- 9213523 TI - [Marfan syndrome]. AB - The author describes three forms of Marfan's syndrome which have only the skeletal symptoms in common. In all instances also ophthalmological symptoms are present but of different types. A familial case was observed only in one family; in the remaining two frust forms of Marfan's syndrome are involved. PMID- 9213524 TI - [An open and controlled study of effectiveness and tolerance of Livostin eyedrops]. AB - The authors present the result of an open controlled trial of the effectiveness of a new antiallergic ophthalmological preparation, Livostin, for local use, a product of Janssen-CILAG. They evaluate the results obtained in 86 patients with symptoms of allergic conjunctivitis. The preparation was followed up for four weeks, in particular the rate of onset of effect and its duration. Evidence was provided that this preparation is a highly effective local antiallergic drug with a long-term effect. PMID- 9213525 TI - [Clinical experience with Spersadex comp. gtt]. AB - The preparation Spersadex comp. gtt. was tested on three groups of patients (60 patients, 109 eyes) in order to assess its effectiveness in indications outlined by the manufacturer. The preparation has a very favourable antimicrobial, and antiphlogistic action. It proved very effective also in the treatment of chlamydial infections and in the postoperative treatment the combination of the antiphlogistic effect and the antibiotic action proved very useful, though tested only in small groups of patients. Spersadex comp. gtt. is a very effective preparation, well tolerated by patients and it supplements in a useful way the range of eye drops by a new antibiotic-corticoid combination. PMID- 9213526 TI - [Postmortem examination of the ocular fundus to determine the time of death]. AB - The author reflects, based on experience from abroad and her own work, on the cooperation of the ophthalmologist in the team of transplantation workers and in forensic practice which by examination of the ocular fundus after death could contribute to the assessment of the time of death. PMID- 9213527 TI - [Intraocular tumors in children]. PMID- 9213528 TI - Some final responses to Dr. Waugh. PMID- 9213529 TI - Osteoporosis. PMID- 9213530 TI - Osteoporosis. PMID- 9213531 TI - Giardia. PMID- 9213532 TI - Chinese dental journals. PMID- 9213533 TI - The Thomas L. Petty 39th Annual Aspen Lung Conference. Genes and gene therapy. Proceedings. PMID- 9213534 TI - [Symptoms, tumor stage and primary treatment in patients with colorectal carcinoma]. AB - OBJECTIVE: To obtain population-related data, previously not available for Germany, regarding primary symptoms, diagnosis and treatment of patients with colorectal carcinoma. PATIENTS AND METHODS: During 1994, in the catchment area of Bonn (town of Bonn, districts of Euskirchen and Rhein-Sieg), there were 354 cases of newly diagnosed colorectal carcinoma. For all of them initial symptoms, primary diagnosis, cancer stage at time of diagnosis as well as surgical and conservative treatment were prospectively documented. By comparing entries from all relevant hospitals and doctors with those of the pathology institutes of the region, data of about 98% of all these patients were recorded. RESULTS: The average age of the patients was older (median of 69 for men, 72 for women) with more far metastases than in comparable groups reported by German university surgical departments. Only 4.6% of the group had occult blood in the stool as the only symptom. Most of the patients had local or regional metastases (T3-4: 63.9%; N1-3: 41%, respectively). Operations intended to be curative were possible in only 65.9% of the cohort. Adjuvant radiotherapy and/or chemotherapy was given to 11.7%, using current consensus recommendations. PMID- 9213535 TI - [Treatment of a large intracoronary thrombus with urokinase and a chimeric monoclonal platelet aggregation inhibitor]. AB - HISTORY AND CLINICAL FINDINGS: 7 days after an operation for intervertebral disc prolapse a 43-year-old man was referred with the clinical and ECG signs of an acute posterior wall myocardial infarction. INVESTIGATIONS: Creatine kinase (CK) activity was raised to 204 U/I (myocardial-specific isoenzyme CKMB of 23.6 U/I, 11.6% of total) and glutamic-oxalate transferase (GOT) activity to 37 U/I. Emergency cardiac catheterisation, performed 4 hours after renewed onset of precordial pain showed no abnormal findings in the right coronary artery, despite the ECG signs, but a definite filling defect in the anterior interventricular branch, which on intravascular ultrasound was an echo-dense noncalcified structure. TREATMENT AND COURSE: After percutaneous transluminal coronary angioplasty in the area of the obstructing structure a free-floating mass was identified in the proximal part of the anterior interventricular branch, most likely a thrombus. Intercoronary thrombolysis was therefore undertaken with urokinase (bolus of 1 mill. IU) together with the chimeric monoclonal antibody c7E3, which inhibits platelet aggregation by blocking the platelet glycoprotein surface receptor IIb/IIIa. Coronary angiography 12 hours later revealed almost complete dissolution of the previously obstructing mass. CONCLUSION: Combining the platelet aggregation inhibitor c7E3 with a thrombolytic agent is an alternative treatment to the current management of intracoronary thrombi. Intravascular ultrasound is a suitable method for demonstrating angiographically inconspicuous or unclear but pathogenetically significant vessel changes. PMID- 9213536 TI - [Auxiliary liver transplantation for acute liver failure after intake of 3,4 methylenedioxymethamphetamine ("Ecstasy")]. AB - HISTORY AND CLINICAL FINDINGS: An 18-year-old patient had for 6 days been suffering from right upper abdominal pain, weight loss, vomiting and yellow discoloration of the skin. For the preceding 8 months he had been regularly taking 1-2 tablets of "ecstasy" (3,4-methylenedioxymethamphetamine--MDMA) per week, the last 8 days before the onset of the described signs. Physical examination was unremarkable, except for pain on pressure over the right upper abdomen and the jaundice. INVESTIGATIONS: The activities of SGOT (756 U/I), SGPT (1450 U/I). gamma GT (164 U/I) and lactate dehydrogenase (539 U/I) as well as total bilirubin level (7.5 mg/dl) were elevated. The synthesising functions of the liver were impaired (thromboplastin time 47%, fibrinogen 116 mg/dl). Abdominal sonography was unremarkable. All virological tests (hepatitis A, B, C and D; Epstein-Barr virus; cytomegalovirus; HIV 1 and 2) were negative. TREATMENT AND COURSE: The suspected diagnosis was acute liver failure after "ecstasy" intake. The cholestasis and the parameters of liver synthesis and hepatocellular functions deteriorated under symptomatic treatment. 15 days after onset of the first symptoms progressive hepatic encephalopathy occurred and required heterotopic auxiliary liver transplantation (piggy-back technique). 5 months later hepatobiliary sequential scintigraphy demonstrated regenerating of the patient's own liver an atrophy of the transplanted liver. Immunosuppression with cyclosporin A and prednisolone was gradually reduced, and the transplant was removed 6 months postoperatively because of an abscess in it. 11 months after the transplantation liver functions is normal and the patient well. CONCLUSION: In young patients with jaundice of unknown origin toxic hepatitis after "ecstasy" intake should be considered. Auxiliary liver transplantation can lead to regeneration during temporary relief of the patient's own liver. After its function has been restored immunosuppression is no longer needed. PMID- 9213537 TI - [Chronic, pressure-induced compartment syndrome. A differential diagnosis for peripheral arterial occlusive disease]. PMID- 9213538 TI - [Adenovirus gene therapy for liver metastases of gastrointestinal tumors. Development status and future prospectives]. PMID- 9213539 TI - [Relationship between metabolic changes and femur head necrosis]. PMID- 9213540 TI - [The language of medicine]. PMID- 9213541 TI - [Is it possible to treat age-related macular degeneration?]. PMID- 9213542 TI - [Clodronate and myeloma]. PMID- 9213543 TI - [Koro: cultural heritage from Southeast Asia]. PMID- 9213544 TI - [Radiological treatment of recurrent variceal bleeding and ascites formation in patients with liver cirrhosis]. PMID- 9213545 TI - [Facial cutaneous leishmaniasis from a vacation to the south]. PMID- 9213546 TI - [Epidural anesthesia in the treatment of severe neuropathic pain in diabetic patients]. PMID- 9213547 TI - [Is snoring a health risk?]. PMID- 9213548 TI - [Sleep apnea and anesthesia]. PMID- 9213549 TI - [Is the self-setting CPAP machine going to be revolutionary in the treatment and diagnosis of obstructive sleep apnea syndrome?]. PMID- 9213550 TI - [Weight loss and the functioning of the autonomic nervous system in sleep apnea patients]. PMID- 9213551 TI - [The epidemiology of narcolepsy in Finland]. PMID- 9213552 TI - [Increased resistance of the upper respiratory airways during sleep]. PMID- 9213553 TI - [Respiratory disorders during sleep in children]. PMID- 9213554 TI - [Narcolepsy--an unusual cause of abnormal sleepiness in children]. PMID- 9213555 TI - The ELLIPS suite of macromolecular conformation algorithms. AB - This paper describes a series of four programmes for the PC based on ellipsoidal representations of macromolecular shape in solution using Universal shape functions, ELLIPS1 is based on simple ellipsoid of revolution models (where two of the three axes of the ellipsoid are fixed equal to each other). If the user types in a value for a shape function from sedimentation or other types of hydrodynamic measurement, it will return a value for the axial ratio of the ellipsoid. ELLIPS2 is based on the more general triaxial ellipsoid with the removal of the restriction of two equal axes. The user enters the three semi axial dimensions of the molecule or the equivalent two axial ratios and ELLIPS2 returns the value of all the hydrodynamic shape functions. It also works of course for ellipsoids of revolution. ELLIPS3 and ELLIPS4 do the reverse of ELLIPS2, that is they both provide a method for the unique evaluation of the triaxial dimensions or axial ratios of a macromolecule (and without having to guess a value for the so-called "hydration") after entering at least three pieces of hydrodynamic information: ELLIPS3 requires EITHER the intrinsic viscosity with the second virial coefficient (from sedimentation equilibrium, light scattering of osmometry) and the radius of gyration (from light or x-ray scattering) OR the intrinsic viscosity with the concentration dependence term for the sedimentation coefficient and the (harmonic mean) rotational relaxation time from fluorescence depolarisation measurements. ELLIPS4 evaluates the tri-axial shape of a macromolecule from electro-optic decay based Universal shape functions using another Universal shape function as a constraint in the extraction of the decay constants. PMID- 9213556 TI - SOLPRO: theory and computer program for the prediction of SOLution PROperties of rigid macromolecules and bioparticles. AB - Single-valued hydrodynamic coefficients of a rigid particle can be calculated from existing theories and computer programs for either bead models or ellipsoids. Starting from these coefficients, we review the procedures for the calculation of complex solution properties depending on rotational diffusion, such as the decays of electric birefringence and fluorescence anisotropy. We also describe the calculation of the scattering from factor of bead models. The hydrodynamic coefficients and solution properties can be combined to give universal, shape-dependent functions, which were initially intended for ellipsoidal particles, and are extended here for the most general case. We have implemented all three developments in a new computer program. SOLPRO, for calculation of SOLution PROperties, which can be linked to existing software for bead models or ellipsoids. PMID- 9213557 TI - A trimeric, alpha-helical, coiled coil peptide: association stoichiometry and interaction strength by analytical ultracentrifugation. AB - Alpha-helical coiled coils are proving to be almost ideal systems for the modelling of peptide and protein self-association processes. Stable oligomeric systems, in which the stoichiometry is well defined, can be produced by the careful selection of the appropriate amino acid sequence, although the principles behind this are still not fully understood. Here we report on a 35 residue peptide, FZ, synthesized by the solid phase method, which was originally designed to form a dimer, but which, in fact, associates to the trimeric state. A detailed characterization of the associative properties of the peptide has been performed by circular dichroism spectroscopy and, in particular, by sedimentation equilibrium in the analytical ultracentrifuge. The presence of the trimeric state, which is stable even at low peptide concentrations, has been confirmed by various independent methods of analysis for molar mass. The effects of both temperature and of guanidinium chloride on the peptide have been investigated and both found to be peptide-concentration dependent. The unfolding induced by the denaturant cannot be adequately described by a simple, two state monomer-trimer equilibrium. PMID- 9213558 TI - [Megakaryocytic thrombopenic purpura associated with enamel hypoplasia]. AB - Patients with platelet disorders usually bleed into superficial sites such as the skin, mucous membranes, genitourinary tract and gastrointestinal tract caused by thrombocytopenia or thrombocyte dysfunction. Thrombocytopenia is caused by one of three mechanisms-decreased marrow production, increased splenic sequestration, or accelerated destruction of platelets. We report a 9-year-old male patient with thrombocytopenic purpura, dyserythropoetic anaemia, freckles on the skin and nanosomia, resembling Fanconi's (constitutional) anaemia. Although many patients with constitutional anaemia own other bony abnormalities, it was not found in this case. On the other hand, the patient showed enamel hypoplasia. To our knowledge, this association has never been reported before. Laboratory diagnosis, clinical findings, dental abnormalities and treatment are presented in details. PMID- 9213559 TI - [Use of drugs in dentistry I. Introductory study]. AB - The authors summarized 514 questioners on the dental patient medication in the general stomatological practice. In the 10 medication group they count 135 different drugs. The number of different minor analgesics were 28 the medicines for local application 26, and the brand of antibiotics 22. The hungarian dentist use 23 different drugs, 5 antibiotics, 5 medicine for local application. An average of the dentist monthly dosage is 97.6 box of medicine (29 antibiotics, 23.2 minor analgesics, 18.4 local anesthetics, 17.5 local drugs). The 70% of the medicines hungarian origin in the dental practice. PMID- 9213560 TI - [The mental foramen syndrome]. AB - The authors outline an extraordinary rare case of metastatic mandible carcinoma. The examined patient who earlier had a mastectomy due to breast carcinoma showed signs of the foramen mentale syndrome (numbness, pain, swelling) also on the right side after 5 years. The process of the generalized malignant tumour was first indicated by the mandibula metastatic tumour of the foramen mentale syndrome. PMID- 9213561 TI - A re-evaluation of factors influencing the sex ratio of spider monkey populations with new data from Maraca Island, Brazil. PMID- 9213562 TI - George Snell - reminiscences 1969 - 1990. PMID- 9213563 TI - Remembering George Snell. PMID- 9213564 TI - Memories of George Davis Snell (1904-1996). PMID- 9213565 TI - Remembrances of George Snell. PMID- 9213566 TI - Snell's unwritten grant application. PMID- 9213567 TI - A tribute to Dr. George Snell. PMID- 9213568 TI - [Axioms of systematic progress in medicine]. PMID- 9213569 TI - [Planning, not evaluation. The role of medical biometry]. PMID- 9213571 TI - [Methods for evaluating diagnosis]. PMID- 9213570 TI - [Randomization in clinical studies. Empirically founded or only a dogma?]. PMID- 9213572 TI - [Evaluating clinical studies. References for critical literature study]. PMID- 9213573 TI - [Evidence-based medicine. Medicine based on rational principles]. PMID- 9213574 TI - [Toxic megacolon after Helicobacter pylori eradication therapy]. PMID- 9213575 TI - [50-year-old patient with rapid ataxia onset, generalized paresthesia and myoclonus]. PMID- 9213576 TI - [Recurrent infarct during aspirin therapy]. PMID- 9213577 TI - [Anticoagulation for immobilized infarct patients]. PMID- 9213578 TI - [After-care of colorectal carcinoma--value of tumor markers]. PMID- 9213579 TI - [Monckeberg sclerosis--noninvasive blood pressure monitoring]. PMID- 9213580 TI - [Mechanisms of action of glucocorticoids]. PMID- 9213581 TI - [Preventing adverse side-effects of glucocorticoid therapy]. PMID- 9213582 TI - The fight against AIDS. PMID- 9213583 TI - Putting medical students to work. PMID- 9213584 TI - It's suicide out there! PMID- 9213585 TI - New polyhalogenated monoterpenes from the sea hare Aplysia punctata. AB - The sea hare Aplysia punctata from Sancti Petri (Cadiz, Spain) contains four new unusual acetates of linear polyhalogenated monoterpenes (5-8) together with four known cyclic derivatives (1-4). The structures were elucidated by interpretation of spectral data. It is suggested that the new constituents of A. punctata might be biotransformation products. Compounds 6-8 showed significant in vitro cytotoxicity against four tumor cell lines. PMID- 9213586 TI - Cytotoxic acylated Spermidine from a soft coral, Sinularia sp. AB - A cytotoxic acylated spermidine was isolated from a Pacific soft coral, Sinularia sp. and its structure determined by NMR and mass spectral analysis. The acyl portion corresponds to (3E)-5-methyltetradec-3-enoic acid. PMID- 9213588 TI - Resuscitation of passengers on two TWA flights. PMID- 9213587 TI - Evaluation of the antifungal activity of natural xanthones from Garcinia mangostana and their synthetic derivatives. AB - The antifungal activity of several xanthones isolated from the fruit hulls of Garcinia mangostana and some derivatives of mangostin against three phytopathogenic fungi, Fusarium oxysporum vasinfectum, Alternaria tenuis, and Dreschlera oryzae, has been evaluated. The natural xanthones showed good inhibitory activity against the three fungi. Substitution in the A and C rings has been shown to modify the bioactivities of the compounds. PMID- 9213589 TI - BCRA 1 testing guidelines for high-risk patients. PMID- 9213591 TI - 14th Bodensee Symposium on Microcirculation: Small Volume Resuscitation in Head Injury. Bodensee, Germany, June 14-16, 1996. Proceedings. PMID- 9213590 TI - [Analysis of membrane protein movement by single particle tracking and laser-beam gradient force optical trap]. PMID- 9213592 TI - [The effect of light of various wave lengths on suppression of nocturnal serotonin N-acetyltransferase activity in chick retina]. AB - PURPOSE: Effects of white and monochromatic light of various wave length on the night-time retinal serotonin N-acetyltransferase (NAT) activity were examined in chicks. MATERIAL AND METHODS: Male chicks (white leghorn; 3-4 weeks old) were used. Chicks were purchased on the day of hatching. All animals were offered ad libitum access to standard food and water, maintained under an ambient temperature of 27 +/- 2 degrees C, 60 +/- 5% humidity, and exposed to 12 hr light: dark illumination cycle for a minimum of 8-10 days before experiments. The daytime light intensity at the surface of the animals' cages was about 150 luxes. Each experiment was performed at least twice. At the beginning of the fourth hour of the dark phase of the light-dark illumination cycle groups of chicks (four animals per group) were exposed to either white or monochromatic light for 5, 10, 30 or 60 min, and then killed by decapitation. In another set of experiments birds were illuminated for 5 min, returned to darkness for additional 5, 15, 60 or 120 min, and then decapitated. Decapitation was done quickly under dim red light (2 luxes). Retinas were isolated and frozen on dry ice. Exposure of animal to light took place in 25 x 21 cm white plastic chamber. Light produced by 5 W 14 bulb (Osram) was passed through a cotton filter or filtered with glass, narrow band interference filters, 7 nm half-peak band-width. The spectral wave length analysis for each interference filter was performed with the aid of Diode Spectrophotometer and the light irradiance the three wave lengths used was measured with YSI Radiometer. To measure NAT activity, retinas were sonicated in 10 vols (w/v) in the proportion of 100 microliters of ice-cold 0.05 M Sodium phosphate buffer (pH 6.8). The enzyme activity was determined in supernatants of tissue homogenates with the radioisotopic method of Steinlechner with modifications of Nowak. Data were expressed as mean +/- standard error of mean (SEM) and analysed for statistical significance by analysis of variance and Student-Newman-Keuls test. RESULTS: The potency of the tested lights to suppress NAT activity of the retina had the following rank order. white > or = green (548 nm) > > blue (434 nm) > red (614 nm). CONCLUSION: In chicks, the suppression of the nocturnal NAT activity produced by a short 5-min pulse of monochromatic light was partially reversible in the retina. The studied chick tissues were far less sensitive to pulses of monochromatic light than of white light. PMID- 9213593 TI - [Dacryocystorhinostomy together with temporary intubation of lacrimal canaliculi as a method of treatment of lacrimal duct obstruction]. AB - The purpose of the study was to present the results of treatment of lacrimal ducts obstruction. The authors describe the advantages of temporary intubation of lacrimal canaliculus with simultaneous dacryocystorhinostomy. Additionally, they prove the harmfulness for probing the lacrimal ducts in adults and describe reasons of unsuccessful operations of dacryocystorhinostomy. The findings have been based on 44 operations within a four-year period. PMID- 9213594 TI - [Use of the laser-flare meter for early diagnosis of corneal graft rejection and for monitoring therapy]. AB - AIM: The purpose of this study was to noninvasively quantify, with the use of a laser-flare meter, the alterations of the blood-aqueous barrier following penetrating keratoplasty. This could objectively indicate the disruption of this barrier in eyes with early allograft rejection, possibly even before the manifestation of the clinical signs, and would help to monitor the efficacy of the treatment. MATERIAL AND METHODS: We used the laser flare-meter (Kowa FM-500) to investigate alterations of the blood-aqueous barrier following uncomplicated penetrating keratoplasty (PK) and in corneal allograft rejection. Examination was performed in 53 eyes of 50 patients after uncomplicated PK (7 days to 12 months after PK), in 20 normal control eyes and in 8 patients with acute allograft rejection before and during the treatment. The treatment consisted of systemic and topical administration of steroids and in some cases additionally topical immunosuppressants. RESULTS: Mean flare values in the control eyes were 3.9 +/- 1.0 photon counts/ms. During the first two weeks after uncomplicated keratoplasty they were significantly increased (18.2 +/- 5.8) but were postoperatively slowly decreasing in time, nearly reaching the control values within 6-12 months. They were considerably higher in eyes with acute allograft rejection (34.0 +/- 13.1), but gradually diminished to 8.1 +/- 1.2 after successful treatment. CONCLUSIONS: The application of laser flaremetry is useful in the follow-up of patients after perforating keratoplasty, especially after high risk grafts. The method helps to detect quantitatively early allograft rejection and is beneficial in monitoring the effectiveness of the treatment. PMID- 9213595 TI - [Corneal endothelium after photorefractive keratectomy procedures]. AB - PURPOSE: Corneal endothelium cell density evaluation in refractive surgery. MATERIAL AND METHODS: The corneal endothelium cell measurements by means of specular microscopy before PRK as well as, 1 month and 12 months after PRK procedures in 62 eyes have been performed. RESULTS: No statistically significant difference was found comparing corneal cell density before and after photorefractive keratectomy procedures. CONCLUSION: The excimer laser radiation used for correction of refractive errors is safe for corneal endothelium. PMID- 9213596 TI - [Hypertrophy of retinal pigment epithelium--epidemiology, clinical characteristics and differential diagnosis]. AB - The aim of the study is to show the clinical features of hypertrophy of the retinal pigment epithelium. The authors have studied five own cases (all women, mean age 42 years). All patients had a complete ophthalmological examination including fluorescein angiography (the follow-up time was 6 to 12 months). They also had general medical examination to find the possible association with intestinal polyposis with probable malignant transformation (Gardner's syndrome). Although hypertrophy of the retinal pigment epithelium is so far a benign lesion, the patients should be checked up every year. All hyperpigmented lesions of the ocular fundus are serious for clinicians because of the suspicion of their malignancies. PMID- 9213598 TI - [Usefulness of color doppler ultrasonography for imaging orbital varices]. AB - PURPOSE: The lesions are difficult to visualize in image diagnostics. The paper aims at describing abilities of ultrasonography in the assessment of varices of the orbit. MATERIAL AND METHODS: The results of color Doppler examinations in 12 patients with moderate proptosis, in whom the presence of varices was suspected, are presented. RESULTS: The method allowed for identification and evaluation of the flow in the orbital region. The typical appearance of orbital varices and changes of the flow during the Valsalva's manoeuvre are described. CONCLUSIONS: The authors indicate a high usefulness of the method in the assessment of the orbital varices. PMID- 9213597 TI - [Clinical manifestations and treatment of Terson's syndrome]. AB - AIM OF THE STUDY: To present clinical manifestations of Terson's syndrome and the results of treatment of vitreous hemorrhages with the use of intravitreous injections of anti-Rh serum. MATERIAL: 8 patients with disc, retinal, vitreous hemorrhages and abducens paralysis which developed after subarachnoid hemorrhages. Intravitreous hemorrhages were observed in 4 of those patients. RESULTS: Vitreous hemorrhages cleared spontaneously in two patients. In the remaining two ones with no tendency for blood resorption intravitreous injections of anti-Rh serum were performed. Vitreous hemorrhage cleared in all the patients after 5-6 weeks and visual acuity improved to 0.8. CONCLUSION: Intravitreous injections of anti-Rh serum can be the method of choice in the treatment of vitreous hemorrhages in Terson's syndrome with no tendency for spontaneous resorption and without proliferative vitreoretinopathy. PMID- 9213599 TI - [The effect of rapid alternating eye covering on formation of binocular vision in convergent squint treated with a localization method. I. Binocular vision examined in free space]. AB - The authors have examined 35 children aged between 6 and 13 years. They were children with convergent fixed squint (angle: from +4 degrees to +26 degrees) with convergent fixation. The patients were treated in accordance with the localizing method -stage II/III. Visual acuity of the master eye was equal 1.0, and of the squinting eye-from 0.3 to 0.8 after correction. Children were practising 2 x 7-10 minutes for 5 days on an apparatus modelled on the Starkiewicz alteroobturator, looking straight forward and with eyes alternatively covered. They had the binocular vision at long and short distance checked with the use of a test with coloured filters (TF), classical Bagolini test (Bki). In all patients the subjective test was positive. Among 35 examined cases the tests for binocular vision did not change in 17. In 16 patients the image seen with the squinting eye came close to the one seen with the master eye by the average of 2.6 degrees at long distance and 4.3 degrees at short distance. Improvement took place after first exercise (8 children) or after all five exercises (8 children). PMID- 9213600 TI - [Effect of fast alternating eye covering on formation of binocular vision in strabismus treated with a localization method. II. Evaluation based on the behavior of functional scotomas]. AB - The authors have examined the behaviour of functional scotomas in the squinting eye in children treated on the static apparatus modelled after the Starkiewicz alteroobturator. They examined 25 children at the age of between 8 and 14 years with convergent squint with macular fixation. Visual acuity of the master eye was equal 1.0 and in the of the squinting eye from 0.4 do 0.8 in correction. The angle of squint was equal from +4 degrees to +26 degrees. The patients were treated in accordance with the localization method-stage II/III. Children were practising on the apparatus 2 x 7-10 minutes for 5 days. In 12 children out of the 25 under examination the functional scotomas were of the same size after exercises, and in 12 children it decreased from 12% to 40%, on average 24.8%. In 1 child the functional scotoma absolutely disappeared. PMID- 9213601 TI - [A case of Jodassohn-Lewandowsky syndrome]. AB - A case of a 25-year-old female patient with reduction of visual acuity, strabismus convergence with corneal scars and paralimbal neovascularisation of her left eye with skin changes of linear skin naevus type is presented. The diagnosis of Jadassohn-Lewandowsky syndrome was based on characteristic skin and eye disturbances. There are also typical changes in mucous membranes and central nervous system. The treatment is difficult and a certain role may be played by cosmetic and repair surgery. PMID- 9213602 TI - [Mixed tumor of the lacrimal gland (case history over a 20-year period)]. AB - A case of the tumor of the orbita is presented. It caused large exophthalmus and partial damage of orbital bones without any loss in the visual acuity in the 20 year period of increasing. Before operation a tumor of the lacrimal gland or an angioma was suspected. A histopathological examination of the whole removed tumor revealed the presence of mixed tumor cells. PMID- 9213603 TI - [Bilateral neuropathy of the optic nerve as a complication of mumps]. AB - Mumps belongs to children viral diseases. However, it rather seldom has general complications, either of internal organs or of the central nervous system. The authors present a case of a 10-year-old girl who two weeks after an uncomplicated mumps had bilateral optic nerve neuropathy. Her visual acuity was low and visual fields were constricted. The symptoms disappeared after 8 days without any treatment. But because of a possibility of optic neuritis the girl was treated with oral steroids. She is still observed in our clinic and still shows pathological changes in static computer perimetry. PMID- 9213604 TI - [Neurophysiologic basis of optokinetic reflex: present state of knowledge]. AB - The optokinetic nystagmus is a phylogenetically old reflexive reaction of the eyes to the movements of the visual surroundings. Two components can be distinguished: (1) the direct one is considered to represent a cortically transmitted loop through the posterior parietal areas: MT and MST; (2) the indirect component passes through brain stem nuclei. One of them is the pretectal nucleus of the optic tract in which we found neurons related to the velocity of the moving pattern. Since in neurons of the oculomotor nuclei the neuronal activity is related to eye position, integration must take place somewhere in the circuit. The brain stern-mediated indirect component is influenced by the vestibular organs, not only by semicircular canals but also by otoliths. PMID- 9213605 TI - Intellectual Property Rights, Naturally Derived Bioactive Compounds and Resource Conservation. Proceedings of the American Society of Pharmacognosy interim annual meeting. San Jose, Costa Rica, October 20-22, 1994. PMID- 9213606 TI - Biodiversity prospecting and benefit-sharing: perspectives from the field. AB - Searching for new biologically active compounds from natural sources starts, obviously, in the field. Plant, microbial or animal materials to be sought and investigated may be selected through a number of approaches. No matter what selection criterion(a) is (are) used, the first step in obtaining the organism concerned is to undertake field collecting work to search for and to collect the organism. Good knowledge on the ecogeographic distribution and precision in the taxonomic identification of the organism(s) sought are crucial if the field work involves the search for a pre-determined organism or set of organisms. Such knowledge and precision during field work are of secondary importance, however, if the search and collection are based on biodiversity or ethnomedical uses, since accurate taxonomic identification may be made at a later date, in a Museum or Herbarium environment. When an individual or institution from a biotechnologically developed country wishes to obtain indigenous raw biological material from a biotechnologically less developed country, an agreement for the procurement of such raw material may be negotiated. Since the effort to search and develop a biologically active compound(s) from natural sources is a long-term process that involves teamwork between field and laboratory scientists, the success of the endeavor will depend in large part on the continued flow of raw material from the field. Goodwill to maintain such a flow may be achieved through appropriate scientific and monetary compensations, both in real-time and in long term sharing of the benefits of discovery. Only with the prospect of financial return to the supplying country will there be an incentive for the protection of the natural resources towards sustainable use and development, as well as to allow time for continuing explorations and discoveries. PMID- 9213607 TI - Medicinal plant genetic resources and international cooperation: the Brazilian perspective. AB - Brazil is a gene rich country, host to 24% of known primate species, between 10 and 15 million species of insects, and 22% of the world's higher plant species. The debate over how and by whom these resources should be protected has intensified over the last few years due to a growing awareness of the links between sustainable utilization of natural resources, conservation of biodiversity, and economic development. Within this context the pharmaceutical exploitation of natural products for drug development has a prominent place. For a significant portion of Brazilian society, fair cooperation is welcome and can facilitate drug discovery. Nevertheless, the complexity of the consequences of patenting and utilization of natural resources calls for a thorough cost/benefit analysis in order to promote policies that can ensure significant and long term benefits for the country. PMID- 9213608 TI - Rules and regulations on the collection in Cameroon of biological materials for biological testing and drug discovery. AB - Regulations on the exploitation of Cameroon's rich biodiversity have tended to emphasize timber exploitation at the expense of non-timber uses. Materials for scientific research have been marginalized. However, in-service instructions usually provide practical guidelines such as specific charges that researchers must pay for the collection of samples. Even though the current law and text of application are silent on regulations addressing the scientific "prospecting' or exploitation of the forests, it is hoped that a particular text in conformity with the current regulations will provide more specific rules governing the collection, testing and exportation of biological materials for drug testing and discovery. PMID- 9213609 TI - Juridical and sociocultural problems on the definition of a law concerning property, usage and access to genetic resources in Colombia. AB - The property, usage, and access to genetic resources, is today one of the primary topics in international business, as a result of the strategic importance of the resources for the biotechnology industry. Internationally, the sovereignty that each country has over its natural patrimony is recognized. However, the new laws of international marketing have obligated countries in the process of development, such as Colombia, to adopt and copy a concept of intellectual property on living resources that does not have anything to do with the country's sociocultural identity, and sometimes even does not take into account its material enjoyment. The new juridical movement that treats genetic resources as private property produces a cultural conflict between indigenous populations, Afro-Americans and peasants, because for them the genetic resources are an element of community life. In these communities, knowledge is freely transmitted; it is an understanding that they have to conserve their agricultural customs and the relationship that they have with the environment. They do not recognize the term "property' according to patenting laws. These elements have to be considered, respected, and guaranteed in the laws that recognize the genetic resources in the country. On the other hand, not even countries that are pioneers in biotechnological development can adopt a concept about patents that is in agreement with the particularities that the living materials possess. This is obviously the reason for the numerous discussions on the legal interpretation, as well as complicated debates in court. Confronting that situation, there are countries rich in biodiversity, such as Colombia, but which do not have a proper concept and are not economically strong in the international context. These countries have to copy inadequate protection policies that do not take into account all their rights. This paper describes some of the technical, juridical, and sociocultural difficulties which Colombia has to confront, in order to set a guideline on patenting living organisms, and on the access and usage of the genetic resources. PMID- 9213610 TI - [Management policy and utilization of botanical resources in the Dominican Republic]. AB - The Dominican Republic has an extension of 48 442 km2. It occupies the eastern portion of the island of Hispaniola and is the second in size within the Greater Antilles. The floral resources of the Island have been extensively exploited without an adequate management policy. This has led to the destruction of about 90% of the forested lands of the country. The Dominican Republic has about 5600 species of vascular plants, of which 36% are endemic, many of which are in danger of extinction, due to overexploitation and/or destruction of the habitats. In order to protect the biodiversity of the Dominican Republic, 22 nature preserves were created, representing about 13.3% of the national territory. The primary institutions charged with the management and protection of the flora are the General Directorate of Forests, National Directorate of Parks, the National Botanical Garden, and the Department of Wild Life under the State Secretary of Agriculture (SEA). Numerous laws, resolutions and decrees have been promulgated, governing forest resources. As to the wildlife, emphasis is given to the protection of the fauna. At present, there is a technical body, which is structuring a Forest Code. Export of products of the wild flora is regulated by the Wildlife Department under the State Secretary of Agriculture (SEA). This Department also administers regulations for CITES. The Department of Plant Sanitation of the SEA and the Dominican Centre of Export Promotion (CEDOPEX) are also involved in the control of the export of biological materials. PMID- 9213611 TI - Collection of biological materials in biodiversity prospecting in India: problems and solutions. AB - Forests are the chief resource for the collection and exploration of biological materials. The past few decades have witnessed a large scale deforestation in India due to substantial pressures generated by population growth, leading to demand for more land for agriculture, urbanization and industrial activities, in addition to increased demand for fuel wood and timber. This has resulted in the loss of soil cover, habitat destruction, environmental degradation and ecological imbalance. This scenario has created a progressive awareness for the conservation and restoration of habitats and, thus, the declaration of many forest areas into protected zones, such as national parks, biosphere reserves, etc., including the protection of some marine areas, by both the National and State Governments. Normally, permission for biological collecting is not granted in these protected areas. In India, forests are a State subject and grant for collection permission is vested with the State Forest Departments. In the absence of any rules, regulations and guidelines, either from National or State Governments, forest authorities impose their terms and conditions, which are arbitrary and even contradictory at times, in the process of granting collecting permits. A set of new rules to be applied throughout the country is needed. PMID- 9213612 TI - [Forestry Law and the conservation of natural areas and wildlife]. AB - The Forest Law of Ecuador consists of 107 articles, whereas its regulations contain 269 articles. They are related to forestry resources, forestry patrimony protection, forests and vegetation, forest production and benefits, the control and mobilization of the forestry resources, research and capacitation, and the forestry industry protection; to natural areas, wild flora and fauna, their patrimony, conservation, and economic support; and to the violation of the law and its judgment. PMID- 9213613 TI - The position of intellectual property rights in drug discovery and development from natural products. AB - Some practical working definitions and perspectives are offered here as to "intellectual property' (IP) and "intellectual property rights' (IPR), particularly as they may pertain to the essential contributions of the parties to an international cooperation or collaboration in drug discovery and development from natural products. Existing and new intellectual properties are examined particularly in respect to feasible ways in which they may be appropriately recognized and protected. It is further illustrated how such suitable recognition and protection may provide the basis to insure fair and appropriate compensation for contributions of existing intellectual property essential for the creation of new intellectual property. The importance of an adequate mutual understanding, and shared realistic perspective, of the overall process and probability of outcome of any given drug discovery and development venture, is emphasized. PMID- 9213614 TI - Rules and regulations of the Government of Vietnam on collection and exportation of biological materials. AB - Situated in Southeast Asia, with a tropical monsoon climate, Vietnam is covered by tropical rain forests over one-quarter of its surface. Forests have been extremely important in the country's economy and they will be essential in its future development. Forests contribute directly to the economy through the provision of building materials and energy and indirectly through foreign exchange earnings, which amount to about US$200 million annually. Forests play a key role in the conservation of biodiversity. They protect watersheds and thus contribute to flood control and water management in the highly productive delta regions. In order for the forests to contribute to the national economy, strong forest management institutions and proper policies are necessary. Forest land use and exploitation should be strictly controlled, and effective programs must be developed. The potential of the forests can only be realized on a sustainable basis through significant changes in current practices. PMID- 9213615 TI - [Intellectual property rights in Costa Rica in the light of the Biodiversity Convention]. AB - This report analyzes intellectual property rights and acquisition of biological samples in light of the Biological Diversity Convention, with emphasis on Costa Rica. It examines the legal framework which exists for the protection of biological resources in this country, especially evaluating the law regarding protection of biota, which was approved in 1992. This includes information regarding access to genetic resources, and regulation for the aforementioned law. It examines the Biological Diversity Convention which was signed at the Rio Summit in 1992, whose objectives and goals, above all, emphasize the subject of distribution of benefits to be derived from the utilization of biological resources. PMID- 9213616 TI - Research, valorization and exploitation of biological resources for medicinal purposes in the Malagasy Republic (Madagascar). AB - Medicinal plants are widely used for treatment of diseases in Madagascar (the Malagasy Republic). Different types of users, including individuals, researchers, groups of researchers and State institutions use medicinal plants as crude materials either for trade, scientific investigations or export. To preserve these forest products for extended use, Malagasy legislation controls the collection of medicinal plants, especially those destined for export. However, according to the law, products coming from Malagasy medicinal plants are not patentable locally. PMID- 9213617 TI - Interests and policies of the state of Sarawak, Malaysia regarding intellectual property rights for plant derived drugs. AB - Sarawak, on the island of Borneo, is known internationally for its rich rain forests, flora and fauna. Its rain forests, occupying two-thirds of its geographical area shelters 2500 tree species, 5500 flowering plants and over 20 000 different kinds of animals and insects. Such abundance of plants, and in particular, in the variety thereof, have attracted the attention of scientists involved in the field of research into their potential medicinal value. Recent discovery that two species of Calophyllum tree in the rain forests of Sarawak produce active anti-HIV agents, has, no doubt, intensified interest in the State's plant resources for scientific research. PMID- 9213618 TI - Guidelines and policies on collection of biological specimens in the Philippines. Philippine Congress, International Convention on Biodiversity. AB - In October, 1993, 16 months after the United Nations approved the International Convention on Biodiversity held in Rio de Janeiro, June, 1992, the Philippine Congress ratified and adopted the Convention. This is a manifestation of the full support of the Philippines for the principles and policies adopted by the UN body on the conservation of biodiversity, sustainable development of biological resources and equitable sharing of benefits between users and owners of biodiversity resources. The Philippine scientific community has long recognized the need for and importance of a national guideline and policy with regard to the collection of plants and animals in the Philippines for scientific or commercial purposes. A series of consultative meetings were held by representatives of government agencies, non-government organizations, private organizations, academic and private persons concerned with biodiversity conservation to formulate national guidelines that regulate the collection of plant and animal specimens in the country. Guidelines were unanimously adopted by various government agencies and academia and a Memorandum of Agreement (MOA) was signed on September 28, 1990. Very recently a new document was drafted, specifically to serve as a guideline for those who desire to undertake sample collecting in the Philippines for biodiversity prospecting. The document is now being reviewed by government departments and agencies and will be presented to the President of the Philippines for signing as an Executive Order (EO). Once signed, this EO will serve as a national policy for bioprospecting in the country. The Philippines is one of the countries in Southeast Asia that has endorsed the adoption of regional guidelines on the collection of plant and animal organisms for drug development. The ASEAN Agreement on the Conservation of Nature and Natural Resources (1985). The Manila Declaration (1992) and lately, the Melaka Accord (1994), all of which were signed by various countries in Asia, are manifestations of this interest. PMID- 9213619 TI - Biodiversity prospecting in Nigeria: seeking equity and reciprocity in intellectual property rights through partnership arrangements and capacity building. AB - The regulation of genetic materials in Nigeria for the isolation of biologically active compounds and/or their exportation from the country fall under the purview of several government departments and parastatals. In principle, biological resources are considered similar to any other natural resource with different levels of stake holders. Specific restrictions, however, apply to the export of food crops. Nigeria is a traditional society where most of biodiversity belongs to what could be appropriately classified as public domain. It has therefore not been easy to carve out property rights from what is generally regarded as communal resources. Private access and occupancy of land and tenure are derived mainly from rights of membership of kindred groups or as custodian of "family' inheritance. The multi-state federal structure allows for negotiations to be conducted mainly at the level of the various State Government Departments responsible for forest resources, and the Federal Government providing the necessary policy guidelines and regulations. The Bioresources Development and Conservation Programme (BDCP), an international NGO based in Nigeria, has adopted an innovative model for biological prospecting based on establishing strategic partnerships and capacity building. PMID- 9213620 TI - Tanzania's policy on biodiversity prospecting and drug discovery programs. AB - Tanzania is endowed with a rich natural resource. The biodiversity comprises over 10,000 plant species, a rich fauna and marine resources. The collection of biological resources is guided by a set of formalities, namely: (i) entry visas, (ii) research permits, (iii) designation of a relevant collaborating host institution or organization, and (iv) eventual joint field work with the collaborating host institution. A facility exists whereby an institution within the country can collaborate with a technologically developed institution or country through mutual research agreement in short and long-term programs. In such a case, Tanzanian scientists collect and export biological materials for drug testing by the collaborating partner. The research agreement is based on the understanding that benefits of the discovery are shared among all parties, namely, the host country's collaborating institution(s), government and indigenous cultures. Rules and regulations governing biodiversity prospecting fall under the categories of (a) floristic resources, (b) fauna or animal substances and (c) marine biological resources. The collection and export of CITES-listed organisms is governed by a separate policy package. Various national institutions are directly involved in overseeing the implementation of these policies. In this paper, these are reviewed to accommodate new conventions concerning research management policies on biological diversity and the sustainable utilization and conservation of these resources. PMID- 9213621 TI - Australian deliberations on access to its terrestrial and marine biodiversity. AB - The predominantly developed country business principle that the natural resource is effectively free, or of very low monetary value, has been significantly challenged in recent years, not only through the recognition of the accelerated rate of depletion of native forest resources and of the space and food demands of increasing populations, but also through international conventions which deal with a wide range of topics from the rights of indigenous people to the Law of the Sea Convention. Australia, classified as a developed country, but located in a geographic region of many developing countries, has, in the past 25 years, demonstrated particular concern for the rights of the people of those countries, as well as for the rights of indigenous people of Australia. The practical international aspects were clearly exemplified in the time, from 1985, when the Australian Institute of Marine Science (AIMS) negotiated, within the National Cancer Institute (NCI) contract, that collections of biological samples in developing countries would be accompanied by an agreement to provide benefits arising from field work, and from any commercial product developments, to those countries. Australia, as a signatory to the Convention on Biological Diversity (Appendix I), continues to analyze the challenge presented by the need to freely exchange genetic resources of common value, e.g. food crops, while insuring an appropriate reward to developing and developed countries, should discoveries be made from their biological resources, which lead directly or indirectly, to high value commercial non-food products. The Prime Minister's Coordinating Committee on Science and Technology established a special working group to recommend on access to Australia's biodiversity. The report arising from the study, and other related issues, are discussed. PMID- 9213623 TI - United Nations Convention on Biological Diversity. PMID- 9213622 TI - Camptothecin and taxol: from discovery to clinic. AB - Camptothecin (CPT) and taxol are secondary metabolites found in the stembark of Camptotheca acuminata, a native of China, and Taxus brevifolia, found in the northwest Pacific coastal region of the USA, respectively. The compounds were isolated through bioassay-guided fractionation of various extracts and through chromatographic fractions. Their unique and hitherto unknown structures were elucidated by nuclear magnetic resonance, mass spectrometry and X-ray analysis. Both compounds have unique mechanisms of antitumor activity; CPT uniquely inhibits an enzyme, topoisomerase I, involved in DNA replication, while taxol binds to a protein, tubulin, thus inhibiting cell division. Taxol has been called the best new anticancer agent developed from natural products, showing particular efficacy against ovarian cancer. CPT and analogs singly or combined with cisplatinum show efficacy against solid tumors, breast, lung, and colorectal, which hitherto have been unaffected by most cancer chemotherapeutic agents. PMID- 9213625 TI - ASEAN Agreement on the Conservation of Nature and Natural Resources, Kuala Lumpur, Malaysia, July 9, 1985. PMID- 9213626 TI - Code of Ethics for Foreign Plant Collectors, developed at the Botany 2000 Herbarium Curation Workshop held in Perth, Australia, October 13- 19, 1990. PMID- 9213624 TI - Natural products research: perspectives from a major pharmaceutical company. AB - Natural products research continues to provide a tremendous variety of lead structures which are used as templates for the development of new drugs by the pharmaceutical industry. Advances in bioassay technology and in chemical methodology have combined to make natural products a cost effective source for new leads. While microbial products have been the mainstay of industrial natural products discovery, in recent years phytochemistry has again become a field of active interest. Drug discovery programs based on microbial products and phytochemicals are discussed and contrasted. PMID- 9213627 TI - The Manila Declaration concerning the ethical utilization of biological resources. Seventh Asian Symposium on Medicinal Plants, Spices and Other Natural Products. PMID- 9213628 TI - The Melaka Accord. Eighth Asian Symposium on Medicinal Plants, Spices and Other Natural Products. PMID- 9213629 TI - Natural product source material use in the pharmaceutical industry: the Glaxo experience. AB - Glaxo PLC has had a significant involvement with Natural Product Source Materials for all of its commercial history and, most recently, has pursued this interest by use of such materials as templates for new lead discovery. Through the expertise and facilities in its Natural Products Discovery Department, Glaxo extracts relatively small quantities of plant material (typically 200-250 g dry weight) and cultures microorganisms from environmental samples (typically 10-50 g). Extracts and fermentation broths are screened in order to detect bioactive principles (BPs). If the potency, selectivity and specificity of the BP is acceptable, isolation, purification and structural elucidation follows. It is most unlikely, in our experience, that the BP itself will become a drug; it is much more likely that we shall need to initiate a medicinal chemistry synthesis program in order to try to produce a molecule that has both the essential biological and desirable chemical properties to become a drug development candidate. This synthetic process is often a long one and our confidence that such a process is worth undertaking is greatly improved if the BP is novel. An essential component of any medicinal chemistry strategy is that it allows us to obtain secure intellectual property rights through patents. Acquisition of product claim protection, the strongest form of patent protection, is of great importance. Safety testing and clinical development of the candidate drug can take 7-10 years, and often more, during which patent protection is constantly eroding. Recognizing that acquisition of Natural Products Source Materials is an issue of growing concern, Glaxo Research and Development Ltd. (GRD), in the early part of 1992, implemented a policy for plant supply. This policy was subsequently modified to embrace source materials such as environmental, soil and marine samples from which fungi, micro- and microorganisms may be obtained. As a direct consequence of this policy, Natural Product Source Materials supply agreements are only concluded with national and international organizations who possess the expertise to identify and collect the samples. It is equally important that our suppliers have the authority, which must be provided to GRD in writing, to collect such materials and to provide them to GRD for extraction and screening purposes. Such materials must be from sustainable and accessible sources. We will not seek to collect any endangered species. Though ethnomedical information can be helpful, it is not essential. Plants must be taxonomically classified. We reimburse the supplying institute for their efforts and their expertise, and recognize an obligation to offer a royalty to the institute in the event that drug discovery, with subsequent commercialization, owes its origin, however indirectly, to a material that it provided. In discussions with the institute, we insist that "a fair proportion' (>40%) of that royalty be used for the direct benefit of the people in the collection source area. In this context, GRD recognizes the importance of local training and education. PMID- 9213630 TI - Biological diversity, indigenous knowledge, drug discovery and intellectual property rights: creating reciprocity and maintaining relationships. AB - When new plant-derived therapeutics based on indigenous knowledge are being explored, it is important that the pharmaceutical companies return benefits to the native populations and the local governments from which the research material was obtained. When a potentially marketable plant product is being developed, it is essential that equitable agreements have already been established between the pharmaceutical companies and the people and/or countries from which this indigenous knowledge was acquired. Equally important is the commitment to provide immediate reciprocity that will enhance the welfare, the biocultural diversity and the well-being of the forest peoples. These measures should commence when a research project begins and continue during its duration. The development of these measures must be based upon the expressed needs of the indigenous communities. The relationship between the stability of the rain forest biocultural diversity, the creation and development of agro-forest resources and the long term benefits to the forest people is highlighted. Examples of initiatives taken by Shaman Pharmaceuticals Inc. and the Healing Forest Conservancy are described and discussed in the context of exploring appropriate use of intellectual property law to address the ethical issues facing all business and research groups working in the tropics. PMID- 9213631 TI - Academic research: policies and practice. AB - The Bayh-Dole Act of 1980 allowed universities in the US to own and manage inventions obtained using federal funds. This Act laid the foundation for university technology transfer activities in most major research universities of the country. Consequently, the interaction between universities and industry has increased, and so has the sophistication in university intellectual property management. UIC's model for managing its intellectual property is efficient and successful, and is the one increasingly used by university technology transfer offices. The model deals with all aspects of UIC's intellectual property: disclosure, protection, marketing, negotiating and licensing, as well as intellectual property provisions in UIC's research agreements, material transfer agreements, option agreements, licensing agreements and others. In a pioneer effort, UIC has developed a policy on contracts for collecting natural product samples for drug discovery which includes royalty sharing and other important provisions. Accompanying this policy we have also drafted a standard contractual agreement for collectors. PMID- 9213632 TI - Gene co-ops and the biotrade: translating genetic resource rights into sustainable development. AB - The 1992 Convention on Biological Diversity marks a basic change in the international status of genetic resources. Prior to the Convention, these resources were considered to be the "heritage of mankind.' Although the intent of this open access regime was to ensure the widespread availability of genetic resources for agriculture and industry, commercial use of the resources provided no additional economic incentive for conservation by source countries. The Biodiversity Convention corrects this policy failure by establishing that states have sovereign rights over their genetic resources, thereby enabling market incentives to complement various multilateral mechanisms that might directly fund biodiversity conservation. A number of obstacles face countries that are translating this broad right to regulate access into specific policies, laws, and regulations designed to meet conservation and development objectives. A review of these obstacles and of trends in technological development suggest that nations and developing country institutions should take a set of actions to develop access legislation and Material Transfer Agreements, establish biodiversity "cooperatives' and intermediary institutions to facilitate information exchange, develop minimum standards for access legislation, and require that prior informed consent of local communities be obtained by all biodiversity collectors. PMID- 9213633 TI - Legal issues in sharing the benefits of biodiversity prospecting. AB - The National Cancer Institute (NCI) is the US Government's principal agency for research on the prevention, diagnosis and treatment of cancer. A critical component of the Institute's mission is the identification and development of new and promising treatments for cancer and AIDS. For many years these efforts have included a program to investigate natural products for potential new therapeutic agents. In 1986, with the advent of new screening techniques, the National Cancer Institute stepped up its exploration of natural products and began world-wide collections of plants in tropical and subtropical regions. In recognition of the principles of the Biodiversity Treaty, NCI appreciates that continued access to the natural products of these countries depends on the Institute's ability to recognize the contributions of these source countries and their indigenous peoples, and to provide them adequate incentives to conserve their natural resources for the purposes of drug discovery. Accomplishing this goal presented several legal issues for the National Cancer Institute. As an agency of the US government, the NCI has an adjunct statutory mission to facilitate the transfer of technology developed through the Institute's programs into the private sector for further development and commercialization, and NCI operates under a national policy to use the patent system to transfer Federally supported research to the private domestic sector. Reliance on patent law may limit the Institute's ability to recognize the rights of source countries and their indigenous people and provide compensation for their contributions. However, other legal instruments, such as contracts, can serve as interim measures to provide compensation to source countries and indigenous populations. The National Cancer Institute's Letter of Collection agreement (LOC, formerly the "Letter of Intent'), is an example of an alternative means that "fills-in the gaps' created by patent law and through which source countries may share in the benefits of natural product development. PMID- 9213634 TI - Workshop on chronic lymphocytic leukemia and hairy cell leukemia. Results of a consensus conference of the German CLL Cooperative Group. Peer-reviewed proceedings. Berlin, Germany, February 7-10, 1996. PMID- 9213635 TI - Acute Leukemia Forum '96. Biology and therapy of ALL in adults: bone marrow transplantation and other approaches. Symposium proceedings. November 8, 1996, Tempe, Arizona, USA. PMID- 9213636 TI - [Nosocomial infection at an intensive care unit: multivariate analysis of risk factors]. AB - BACKGROUND: Nosocomial infections, especially in the intensive care unit, are a very important problem due to their frequency and important consequences (morbility and mortality). On the other hand there are some risk factors and some preventive measures which are involved in the appearance of the nosocomial infections. The purpose of this work was to recognize these risk factors and to identify the preventive measures which are effective, and also to quantify the participation of each risk factors/preventive measures in the development of the nosocomial infections. PATIENTS AND METHODS: Follow-up of a cohort of patients admitted to the intensive care unit of the General Hospital of La Paz (Madrid, Spain) during a year and with a stay of at least 48 hours. RESULTS: We have found a cummulative incidence of patients with nosocomial infection of 32.8%. More than 80% of the patients received antibiotic treatment during their stay in the intensive care unit. The stay of the patients no infected was 4 days while the stay of infected patients was 20 days. We have found a mortality of 29.5%, which was greater in the patients who were infected (42%). In the multivariate analysis we have developed an equation to predict the development of the nosocomial infection. The following variables were identified: six or more instrumentations (OR, 4.75; 95% CI, 2.75-8.19), more of ten days of hospitalization previous to the appearance of the first nosocomial infection (OR, 4.17; 95% CI, 2.60-6.70), administration of muscle relaxing drugs (OR, 2.25; 95% CI, 1.43-3.55), nasogastric tube (OR, 2.19; 95% CI, 1.25-3.84), and altered consciousness (OR, 2.19; 95% CI, 1.25-3.84). Therefore, those patients who present some of these characteristics should be monitored in a special way due to their high risk of development of a nosocomial infection. CONCLUSIONS: Several factors play an important role in the development of a nosocomial infection in the intensive care unit; these are not only intrinsec (especially the altered consciousness) but also extrinsec (instrumentations and drugs), as well as the stay at the hospital previous to the appearance of the first nosocomial infection. PMID- 9213637 TI - [Analysis of reliability of 3 portable blood glucose dosimeters. Comparison of various methods for the study of feasibility of clinical parameters]. PMID- 9213638 TI - [Tuberculosis outbreak: significance of exposure time versus proximity to infection source]. AB - BACKGROUND: The polymorphic length restriction fragment technique (PLRF) complements epidemiologic investigations. The aim of this study was to present an outbreak of tuberculosis in which the risk factors of infection and disease were studied. PATIENTS AND METHODS: A descriptive study was carried out in cases of tuberculosis. The isolated strains of Mycobacterium tuberculosis were typed by the PLRF technique. A study of the prevalence of infection was carried out among the 61 classmates of two classrooms (A and B) which the index case attended. An historical cohort study was thereafter performed among the cases with infection. The association of the dependent variable (infection or tuberculosis disease) with the remaining variables was determined by the odds ratio (OR). RESULTS: The incidence of disease was 9.8% (6/61). The strains isolated in 5 patients presented the same PLRF pattern. The prevalence study detected 28 infections (45.9%). Five cases (17.9%) were detected on the second tuberculin test. By multivariate analysis showed that the hours of exposure (1.8-3.2 hours, OR = 2.0; > 3.2 hours; OR = 10.2) were the risk factor for infection. The BCG vaccine, the intensity of the reaction to the tuberculin test and age could be associated with the risk of disease. CONCLUSIONS: The focus of the outbreak was confirmed by the PLRF technique. The importance of repeating the tuberculin test in whom the test was negative on the first test is of note. To evaluate the risk of infection the time of exposure is more important than the proximity to the index case. PMID- 9213639 TI - [Methicillin-resistant Staphylococcus aureus infection. Example of the current complexity of hospitals]. PMID- 9213640 TI - [15 years of research on toxic oil syndrome]. PMID- 9213641 TI - [Corneal ulcer caused by Nocardia asteroides after penetrating keratoplasty]. AB - We report the case of a 43-years-old patient, to whom a corneal transplantation was made because he presented a Salzmann nodular degeneration in his left eye. The patient was observed every week and his development during the following months was good. Nine weeks later he was attended at the emergency room of the hospital, with an intensive secretion and partial loss of vision in the operated eye. It was detected a peripheral ulcer of a diffused borders with a loss of epithelium and anterior stroma in the superior temporal part of the cornea. Five days later, the microbiological cultures confirmed the presence of Nocardia asteroides. In spite of the initial good evolution of the ulcer treated topically with a 20% sulfacetamide and trimetoprim-sulfadiacine p.o., the graft ended unsuccessfully. PMID- 9213642 TI - [Oral drug administration]. PMID- 9213643 TI - [Blood lead levels in a population of Fuenlabrada, Madrid]. PMID- 9213644 TI - [Mortality caused by human immunodeficiency virus in the Autonomous Communities in 1992]. PMID- 9213645 TI - [Hospital location and the contract of drug clinical trials]. PMID- 9213646 TI - [Patterns of resistance to antitubercular agents in Girona. Diagnostic and therapeutic implications]. PMID- 9213647 TI - [Aprotinin in orthotopic liver transplantation]. PMID- 9213648 TI - [Sweet syndrome associated with essential thrombocytopenia]. PMID- 9213649 TI - [The correct translation of an anglicism]. PMID- 9213650 TI - [Stress and burnout among physicians. Similar problems in different health care systems]. PMID- 9213651 TI - [Future structure of the Medical Society. Members must have the opportunity to influence the society's policy]. PMID- 9213652 TI - [Johan Calltorp on the health care crisis: politicians are at loss--a good occasion for the physicians to take initiative!]. PMID- 9213653 TI - [Who listens when the personnel remains silent?]. PMID- 9213654 TI - [Alarming reduction of specialist education]. PMID- 9213655 TI - [Views on the National Diabetes Registry]. PMID- 9213657 TI - [Children are sick in the psychiatric world]. PMID- 9213656 TI - [An objective dialogue on private care costs is needed]. PMID- 9213658 TI - [The Ersta hospital's help during the Estonia disaster must not be forgotten]. PMID- 9213659 TI - [Abortion registry--don't!]. PMID- 9213660 TI - [The Dagmar funds were correctly used in Varmland]. PMID- 9213661 TI - [To clone or not to clone...]. PMID- 9213662 TI - [Who should perform thyroid surgery? Recommendations of the National Board of Health and Welfare]. PMID- 9213663 TI - [Treatment of Graves disease. An endocrinologic-surgical perspective]. PMID- 9213664 TI - [Programmed cell death is a condition for life. When the balance is upset, disease appears]. PMID- 9213665 TI - [Warning on antidepressive agents during pregnancy. Even low doses can affect the infant]. PMID- 9213666 TI - [Are laboratory results reliable? Serious factors are involved]. PMID- 9213667 TI - [Blood is thicker than water. More people want to donate an organ to their relatives than to unknown persons. Similar situation exists when it comes to receiving an organ]. PMID- 9213668 TI - [1,4 millions Swedes are on the donation registry. Still several thousands newly registered persons per month]. PMID- 9213669 TI - [A new report from the SBU on treatment with neuroleptics. Over 2000 scientific articles critically scrutinized]. PMID- 9213670 TI - [More unified registration in two databases. Easier exchange of esperiences on the cause of medical errors]. PMID- 9213671 TI - [Are mortality figures from the National Board of Health and Welfare acceptable as a quality indicator in myocardial infarction?]. PMID- 9213672 TI - [On the border to Norway...A medical folklore scientist with a passion for working with patients]. PMID- 9213673 TI - [My three mothers--on children's right to know their origin]. PMID- 9213674 TI - [Statistical data as scientific cosmetics]. PMID- 9213675 TI - [Shouldn't group accident insurance cover all kinds of accidents?]. PMID- 9213676 TI - [Treat sinusitis as before!]. PMID- 9213677 TI - [Central nervous system stimulants to hyperactive children? Time to soften up the attitude towards an efficient treatment]. PMID- 9213678 TI - [Glimpses from four different primary health care ditricts: increasing demands, worsened working conditions]. PMID- 9213679 TI - [It's an art to find beauty of simple things]. PMID- 9213680 TI - [News about zoonoses. International cooperation is necessary for development of vaccines, vector control and education]. PMID- 9213681 TI - [Heart surgery with reduced occurrence of infections. New routines and continuous registration are proposed]. PMID- 9213682 TI - [Measurement of folic acid and vitamin B12 in the elderly. Low serum levels result in worse memory]. PMID- 9213683 TI - [An American scholarship for surgeons]. PMID- 9213684 TI - [Dare to perform the surgery vaginally! Vagina hysterectomy is to be prefered when there is no indication for the abdominal intervention]. PMID- 9213685 TI - [Epidemic C difficile-associated diarrhea is a reality. Fewer prescriptions and more soap]. PMID- 9213686 TI - [Investigation of epilepsy. Magnetic tomography more and more important as a diagnostic technique]. AB - Electroencephalography (EEG) and neuroradiology are both indispensible techniques in cases of suspected epileptic seizure, when the aim of investigation is to determine whether the seizure was of epileptic nature, and if so whether it was the result of specific provocative factors or an expression of epileptic disease. In the latter case, the epileptic condition should be classified and its aetiology determined, if possible. Routine or sleep EEG providing interictal epileptiform discharges is a useful aid to differential diagnosis. To obtain EEG recordings during actual seizures, long-term recordings, using either ambulatory equipment or an EEG-video procedure, are usually used. The combination of EEG and video recording, using surface or surgically implanted electrodes, is a procedure of major importance in the evaluation of patients refractory to medical treatment and possible candidates for epilepsy surgery. In cases of epilepsy suspected to be caused by tumour or cerebrovascular disease, neurological investigation does not differ from that routinely used in such conditions. MRI (magnetic resonance imaging) techniques have become important aids in the preoperative work-up in cases of chronic therapy-resistant partial epilepsy. MRI has also simplified the identification of minor morphological abnormalities causing partial epilepsy, and is the method of choice in such cases. The sensitivity of MRI is improved by its combination with volumetric measurements and spectroscopy. The use of functional neuroimaging with SPECT (single photon emission computed tomography) and PET (positron emission tomography) during seizures provides further information. A promising new development is the co-registration of MRI and functional imaging (dipolar reconstruction of EEG spikes and seizure patterns, SPECT, PET). MRI is a cornerstone of the preoperative work-up, but diagnosis and the choice of therapeutic approach is always based on the clinical picture, EEG, and functional and morphological imaging. PMID- 9213687 TI - [A case report and medical history. 100 years old and one kidney since 80 years back]. PMID- 9213688 TI - [How often do we use the cortisone injection? Risk of overconsumption in musculoskeletal pain conditions]. PMID- 9213689 TI - [The 50th anniversary of para-amino-salicylic acid--a historical retrospect. Again and important agent in tuberculosis]. PMID- 9213690 TI - [Develop "silent knowledge" and learn from other clinics. Successful way for quality work within the psychiatric sector]. PMID- 9213691 TI - [Discussion on ethics in France. Scietists want the embryo research to be permitted]. PMID- 9213692 TI - [Working abroad?]. PMID- 9213693 TI - [Accredited laboratory? Worthless information in a scientific article]. PMID- 9213694 TI - [Widen the time perspective in the debate on euthanasia! What's the opinion of the clinically active?]. PMID- 9213695 TI - [Irrelevant analysis in an editorial of the diagnosis registration on prescription forms]. PMID- 9213696 TI - [Explicit signals may restore patients' confidence for the AT-physicians]. PMID- 9213697 TI - [Constructive proposal on improvement of emergency medical training is wanted]. PMID- 9213698 TI - [What is happening with the institution for drug information scrutiny?]. PMID- 9213699 TI - [Erroneous conclusions on diagnostics of Chlamydia]. PMID- 9213700 TI - [Public health in a perspective of the EEC]. PMID- 9213701 TI - [Gonorrhea is increasing among homosexual men. Is the risk of STD and AIDS forgotten?]. PMID- 9213702 TI - [Ultrasound is ideal in hip physiolysis]. PMID- 9213703 TI - [A study of handicapped children demanding prolonged care. Their survival depends on technology]. PMID- 9213704 TI - [Technology-dependent children with prolonged care. Cooperation within home care services prevents parents' stress]. PMID- 9213705 TI - [Cytogenetic diagnosis of tumors. Clinical significance of chromosome aberrations]. PMID- 9213706 TI - [Surgical treatment is efficient in epilepsy]. AB - Epilepsy surgery has a very long tradition, and recent advances in diagnostic and surgical procedures have enabled a number of patients with drug-resistant epilepsy to be treated successfully. In addition to the conventional clinical work-up, candidates for epilepsy surgery undergo evaluation by a multidisciplinary team using a battery of neuroimaging and neurophysiological procedures. Such teams have been established at all six university hospitals in Sweden. PMID- 9213707 TI - [Guidelines for economic evaluation of drugs. Something for Sweden?]. PMID- 9213708 TI - [Disturbed thermostat causes problems during climacteric]. PMID- 9213709 TI - [Acupuncture as an alternative to estrogen. New therapeutic methods for climacteric problems are tested]. PMID- 9213710 TI - US patients do not always get the best end-of-life care. PMID- 9213711 TI - Osteoporosis drugs show early promise. PMID- 9213712 TI - Avoiding stomas with total anorectal reconstruction. PMID- 9213713 TI - [Prevalence of atopy indicators in the adult population of the Zagreb region]. AB - In a sample of 583 subjects (303 men and 280 women) intradermal test with 13 inhalatory allergens, total serum IgE (PRIST Phadezym) determination and nonspecific bronchial reactivity measurement by histamin test, were performed. History data on respiratory symptoms was also taken. The following were evaluated: mean urtica diameter D+d/2 > 8 mm (KT+), value of IgE > 125 IU/ml, and bronchial hyperreactivity (BH), separately in symptomatic and asymptomatic subjects. In symptomatic smokers total IgE (G.M. IU/ml) was higher in women than in men (177.70:68.20-p < 0.01), and in asymptomatic smokers it was higher in men than in women (50.71:33.79-p < 0.01). A larger number of allergens discriminated skin reactivity between the symptomatic and asymptomatic women (grass, tree and weed pollen-p < 0.001; house dust-p < 0.05), than between the symptomatic and asymptomatic men (grass pollen-p < 0.001). The level of IgE < 125 and > 125 IU/ml discriminated significantly the prevalence of persons with KT+ in the group of symptomatic men (80%:27%-p < 0.001) but not in the group of symptomatic women (77%:67% N.S.). BH only partially coincides with KT+ and raised IgE (> 125 IU/ml) in symptomatic subjects (men 20%, women 23%) and in asymptomatic subjects only in men (8%). PMID- 9213714 TI - [Aneurysms of the extracranial part of the carotid artery]. AB - The purpose of this article is to take firm position on the surgical treatment of the carotid artery aneurysm on the basis of ten years of experience, considering their rare occurrence and significant pathology. From January 1984 to December 1994, ten patients with aneurysms or pseudoaneurysms of extracranial carotid arteries were diagnosed and operated in the Department of Vascular Surgery, Clinical Hospital "Sestre milosrdnice" in Zagreb. In the same period, eight hundred operations of extracranial carotid arteries were performed. Special emphasis is put on the etiology of the disease where atherosclerosis is prevalent, but cases of traumatic, mycotic and postoperative aneurysms are also shown. In symptomatology, signs of palpable local tumor dominated in six patients, while in three patients central nervous system symptoms were found. All patients with neurologic symptoms were in the group with atherosclerotic aneurysms. In nine patients resection of the aneurysm and reconstruction with the interposition of a part of vena saphena magna or with allograft was performed. In one patient, neoanastomosis of the internal carotid artery in the common carotid artery was performed following the resection of a small aneurysm. In the early postoperative period there was no morbidity nor mortality. In view of frequent preoperative neurologic complications and good postoperative results, surgical treatment is indicated in all cases. Reconstruction is always indicated, and the procedure of choice is reconstruction with the interposition of a part of great saphenous vein. PMID- 9213715 TI - [Bronchoscopy in primary pulmonary tuberculosis in children]. AB - In the period from January 1, 1991, to June 30, 1994, 204 children (aged 0-14 years) with primary lung tuberculosis were treated in our hospital. Bronchoscopic examinations were done in 112 (54.9%) patients. Positive bronchoscopic finding was observed in 44 (39%) patients, and in three out of them there was a penetration inside the bronchi. This condition was cured within one year of treatment. PMID- 9213716 TI - [Intravenous magnesium in acute myocardial infarct]. AB - The effects of magnesium (Mg) on the incidence of arrhythmias and on mortality were evaluated in 61 patients with documented acute myocardial infarction (AMI), in a randomized, double blind placebo controlled study. During the first 24 hours 31 patients received infusion of 1000 ml isotonic saline with 17 g MgSO4 and 30 ones received only equal volumes of saline as placebo. The baseline characteristics of the population including serum Mg and potassium were similar in the two groups. Severe arrhythmias were not as frequent in the Mg group (22%) as in the placebo one (36%), but the difference was not statistically significant (p = 0.05). There is no significant difference between the groups neither in mortality (3.2%; 0%) nor in conduction disturbances (3.2%; 3%). The adverse effects of Mg therapy were transient and therapy interruption was not required. CONCLUSION: Although the trial is not an extensive one, and the results are nonsignificant, we consider intravenous Mg therapy to be simple and cheap with its place in the treatment of AMI in properly selected patients. According to available data it should be administered as soon as possible after the onset of AMI symptoms, before thrombolytic therapy, in 24-hrs. dose not exceeding 50-65 mmol probably through several days, but the optimal duration of therapy should be further investigated. PMID- 9213717 TI - [Cotrimoxazole in the treatment of Wegener's vasculitis (case report)]. AB - A female patient suffering from Wegener's vasculitis of upper respiratory tract, lungs, kidney and skin, is presented. The diagnosis has been established on the basis of clinical course, histopathologic examination of lung tissue obtained by transbronchial lung biopsy and positive serum antineutrophil cytoplasmatic antibodies (ANCA). The patient was successfully treated with trimethoprim sulfamethoxasole (daily dose 320 + 1600 mg) leading to complete clinical remission. The course of disease and the effects of treatment were strongly drug dose-related. We emphasized some diagnostic difficulties and new trends in the therapy of Wegener's granulomatosis and other ANCA positive vasculitides. Possible mechanisms of the trimethoprim-sulfamethoxasole efficiency in Wegener's granulomatosis are also discussed. PMID- 9213718 TI - [Which patients should be treated with plasma exchange? Indications for plasma exchange therapy]. AB - What is the current opinion on therapeutic plasma exchange, after 20 years of clinical use for numerous patients and disease states? Principles of rational therapy have broken through into the field of therapeutic plasmapheresis during the last decade. Efficacy proven by controlled clinical trials is the prerequisite for determining indications. Superiority of plasma exchange over other therapeutic modalities has been established for only a limited number of rare diseases. Plasma exchange treatment is still actual for plasmatic hyperviscosity syndromes in the course of haematological and rheumatological diseases. Indications for therapeutic plasma exchange in nephrology are narrowed to some forms of Goodpasture's syndrome, rapidly progressive glomerulonephritis with antineutrophil cytoplasmic antibodies and renal insufficiency, and thrombotic thrombocytopenic purpura/haemolytic uraemic syndrome. Plasma exchange therapy is indicated for severe forms of Guillain-Barre syndrome and myasthenia gravis. Therapeutic benefit of plasma exchange was not found by controlled clinical trials in patients with rheumatoid arthritis, systemic lupus erythematosus, polymyositis, dermatomyositis, multiple sclerosis and rejection of kidney allotransplant. PMID- 9213719 TI - [Comparison of open and laparoscopic appendectomy]. AB - The first laparoscopic appendectomy was performed by Senn in 1982. Since then, the dilemmas about the validity of this operation in relation to open operation have persisted. Many authors presented the technique modifications and results that are very different. The retrospective results, cost, duration of hospital stay and postoperative recovery analyses for fifty patients in each group were done in this study. Laparoscopic operations were done by "two-handed" technique and in different ways of appendix and mesoappendix closing and cutting. Endoscopic linear cutters were used in the second part of the study. When comparing parameters, laparoscopic operation in relation to open operation is equally safe; quicker; with less postoperative pain; with less wound infections rate; with shorter hospital stay; with less staff time involved; with faster recovery and return to work; more expensive; with better cosmetic effect. In conclusion, laparoscopic appendectomy is better, although more expensive, than open operation, so it should be recommended. PMID- 9213720 TI - [A remembrance of Dr. Jaksi Racic]. PMID- 9213721 TI - [Tuberculosis--the epidemiologic situation]. PMID- 9213722 TI - [New Croatian regulations on iodized salt]. PMID- 9213723 TI - [Smokers' edema of the vocal cords]. AB - The purpose of this study was the analysis of the etiology, clinical and phonatory characteristics in the subjects with Reinke's oedema. The research was done on 105 subjects, of whom 95 women and 10 men of the age between 28 and 83. The results show that according to the sex, the frequency of the Reinke's oedema is higher in women than in men. The disturbances, not according to the sex, appear mostly in the age between 40 and 60. The voice disturbances last usually for some years. Most of the subjects were smokers (80%) which confirms that smoking (with great intensity and for many years) is the main etiological factor in the development of Reinke's oedema. Less important are considered the voice loadings of long duration, while allergy and acute respiratory infection are mentioned as the possible factors. The values of the phonatory index show less effective air usage during the phonation in the subjects. From the tridimensional spectral analysis of voice that was done on 44 women and 4 men, the data about the frequency of the fundamental laryngeal tone (f degree) were taken. Analysing these data, the lower values of the frequency of the fundamental laryngeal tone were found. The elaboration with the Student's t-test was obtained a statistically important difference of the mentioned parameter in both sexes among the subjects and persons with normal voice. We can conclude that the alteration of the phonatory function is present and in consequence of that, the voice is not effective in communication and less valorous in the media. PMID- 9213724 TI - [Bronchopneumonia, obstructive bronchitis and bronchiectasis in children with adenovirus infections]. AB - During the five-year period 56 children were treated in hospital due to respiratory infections caused by adenovirus. Clinical, laboratory and radiographic signs of the illness are presented. The infections manifested as the upper respiratory tract infection in 3 patients (5.4%), obstructive bronchitis in 16 patients (28.6%), bronchopneumonia in 32 patients (57.1%) and bronchiectasis in 5 patients (8.9%). Three children were operated: bilobectomy was performed in two cases, and left-sided pulmonectomy in one case. Histologically, bronchiectasis was found in two cases and bronchiolitis obliterans in one case. In this work we tried to show the gravity, importance and consequences of the adenoviral infection of the respiratory tract in children. PMID- 9213725 TI - [Retroperitoneal marsupialization of renal cysts]. AB - The use of minimal invasive surgery in urology continue to increase. Retroperitoneoscopic approach in performing minimal invasive surgery of retroperitoneum shortens the duration of operation in comparison with transabdominal approach, with minimal risk of intraabdominal complications. We described the use of the retroperitoneoscopic approach to the upper pole of a kidney for marsupialization of a symptomatic renal cyst. The procedure was minimally traumatic, morbidity was negligible and the patient was discharged from the hospital the third day after the operation. We believe that retroperitoneoscopic management of giant symptomatic renal cysts will be applicable, together with other existing methods. PMID- 9213726 TI - [Pseudothrombocytopenia caused by ethylenediaminetetraacetic acid (EDTA) (case report)]. AB - Pseudothrombocytopenia is a laboratory artefact that can introduce serious problems in diagnosis and treatment in patients with low platelet count. The most common reason for this artefact is in vitro platelet clumping in blood samples collected into ethilenediaminetetraacetic acid (EDTA) anticoagulant. The clumping activity is greater at temperatures less than 37 degrees C, and the EDTA concentrations required for clumping are 20 times below anticoagulant concentrations. In this article we described the case of a female patient with diagnosed EDTA induced pseudothrombocytopenia. The cause of incorrectly low platelet counts was proved by simultaneous analysis in blood samples collected into EDTA anticoagulant and into heparin as a control sample. Absences of incorrectly low platelet count in heparin sample and rapid decrease of platelet count in EDTA sample were noticed. Decrease in platelet count was accompanied by increase in the number of leukocytes, so called pseudoleukocytosis. Careful examination of blood film is necessary to establish correct diagnosis, promptly after the blood collection and approximately two hours later. It is important to verify formation of clumps two hours after the blood collection and also progressive reduction in the platelet count in EDTA sample. By blood assessment conducted in this concern it is possible to avoid severe misinterpretation in such patients. PMID- 9213727 TI - [Use of angioscopy in vascular surgery bypass with an in situ approach--first year experience]. AB - Femoropopliteal and femorocrural bypass, managed by "in situ" procedure with the use of angioscope, became a standard surgical method in the treatment of occlusive process in the arteries of lower extremities in the Department of Vascular Surgery, Clinical Hospital "Sestre milosrdnice" in Zagreb. The capabilities of angioscopy in diagnosis and in surgery are presented, with special emphasis on bypass "in situ" procedure. Technical difficulties encountered in bypass formation and our solutions are described. From December 1993 to December 1994, eleven patients were operated. Common femoral artery was used for proximal anastomosis in four patients. Superficial femoral artery was used in five patients, and profound femoral artery was used in two patients. The third segment of popliteal artery was used for distal anastomosis in five cases. In six cases crural bypass was done. Three of them were on posterior tibial artery, two were on fibular artery and one was on anterior tibial artery. Two out of four or 50% of femoropopliteal bypasses are patent after the first year. One patient died of cardiorespiratory complications in the early postoperative period. Femorocrural bypasses are patent in five out of six patients or 83.3%. Bypass with great saphenous vein "in situ" is the procedure of choice, especially in femorocrural position. PMID- 9213728 TI - [Initial experience with ventilation-perfusion scintigraphy in patients with a suspected pulmonary embolism]. AB - Ventilation-perfusion (V/P) scintigraphy was performed 62 times in 57 patients suspected of having pulmonary embolism (PE). The aim of this study was to present the results and our first experiences in V/P scintigraphy, as well as to point out some specificities of the study. Perfusion scintigraphy was performed following i.v. administration of 99mTc MAA. If the finding was positive, ventilation scan was performed directly after the inhalation of 99mTc DTPA aerosol. Based on the comparison of both findings the patients were divided into four groups: normal finding (8.1% of patients), low (54.8%), medium (22.6%), and high level of PE probability (14.5%). As V/P scintigraphy is a very sensitive and non-aggressive method, it is our opinion that it should be included in PE diagnosing as a "screening" method, because the scanning results greatly influence further therapeutical and diagnostic treatment of the patient. PMID- 9213729 TI - [Digital photoplethysmography and digital "strain-gauge" plethysmography in differential diagnosis of edema]. AB - Photoplethysmography is a noninvasive method for fast diagnosis of blood flow disturbances in the venous system of the lower limbs. It is suitable for discriminating normal from pathological venous findings. The technique has been evaluated in three groups of subjects with edema of the lower extremities. The value of this screening method in the diagnosis of vascular disorders and edema of the lower limbs is discussed. Venous occlusion plethysmography is used for discriminatory and quantitative evaluation of venous function. The diagnostic value of strain gauge plethysmography was assessed in three groups of subjects. Measured parameters were venous capacity and venous outflow. Digital photoplethysmography is suitable for the discrimination of venous edema of the lower limbs from edema caused by other factors, and digital "strain gauge" plethysmography for the discrimination of edema of the lower limbs, caused by phlebothrombosis from edema caused by other factors. PMID- 9213730 TI - [The simple (simplex) renal cyst--a new (old) problem]. PMID- 9213731 TI - [Routine immunologic tests in atopic dermatitis]. AB - The study comprised 100 examinees with atopic dermatitis and 50 examinees with chronic urticaria (adult patients and children), as well as 50 examinees in a control group. The parameters for monitoring included laboratory findings; values of eosinophiles, total lymphocytes, T and B lymphocytes, serum immunoglobulin (IgG, IgA, IgM) total IgE and specific IgE; results of cutaneous tests: prick and intradermal test to inhalative and nutritive allergens, scratch test to preservatives and additives, and epicutaneous (patch) test to contact allergens. In detection and monitoring of patients with atopic dermatitis, several methods were applied with different diagnostic success. The values of total number of lymphocytes and T and B lymphocytes were mainly within the normal limits, therefore these were not good diagnostic parameters. Eosinophiles were mainly higher and were useful in determination of AD. Mainly normal values of IgE, IgA and IgM were found by radial immunodiffusion, but the ELISA method, as well as the RIST method, showed high values of total IgE. Of the applied skin tests, the most important was intradermal test which gave most of the positive results in the patients with atopic dermatitis (97.7%), then followed scratch test (83.3%), patch test (64.3%), and finally epicutaneous test (57.1%). PMID- 9213732 TI - [Evaluation of the central nervous system activity according to the data of a psychophysiological "Rhythmotest" device in workers engaged in manufacture of construction plastics]. AB - The paper deals with some topical problems in monitoring the central nervous system of workers contacting with chemicals in the manufacture of building plastics. The use of psychophysiological "Ritmotest" devices is demonstrated to enhance the diagnostic efficiency of occupational diseases in this group of patients. PMID- 9213733 TI - [Metrological problems of laboratory diagnosis]. AB - Medical laboratory diagnosis is progressing from one analysis to a group of analysis with rather intricate statistical processing of data via multidimensional analysis, at the same time it is necessary to define a patient's individual normal levels, i.e. a "health certificate". In this regard, the process may be achieved when complex analyzers, which have a reliable metrological support, and systems for collection of both clinical and laboratory information and its simultaneous analysis are put into laboratories' practical work. PMID- 9213734 TI - [The DICOM standard in computer-assisted medical technologies]. AB - The paper outlines one of the promising standards to transmit images in medicine, in radiology in particular. The essence of the standard DICOM is disclosed and promises of its introduction into computer-aided medical technologies. PMID- 9213735 TI - [Method of calculating the probability of radiation-induced complications in organs and tissues in relation to volume of radiation and dose fractionation]. AB - Mathematical models have been first developed, which describe the probability of radiation-induced complications of particular types as a function of the dose and volume of radiation and the scheme of dose fractionation in time. New mathematical models were derived from the synthesis of the mathematical models which describe the probability of radiation-induced complications for the fixed scheme of dose fractionation and which describe the equivalent schemes of dose fractionation for the fixed likelihood of radiation-induced complications. PMID- 9213736 TI - [Operating safety of magnetic resonance tomographs]. AB - The factors determining the operating safety of magnetic resonance (MR) tomographs are considered. It is stated that the main factors to be accounted in defining safety measures are exposures to 3 types of magnetic fields: the static leakage field of the magnetic system of a MR tomograph; the radio-frequency field of RF coils, and the rapidly varying fields of gradient coils. Possible impacts of exposure to the above fields on patients and staff are enlisted. The indices accepted to rate the operating safety of MR tomographs and their maximum acceptable levels are given. PMID- 9213737 TI - [Resolutions of the Scientific-Technological Conference on Biomedical Engineering, Biomedical Equipment-96. Moscow, 8-10 October, 1996]. PMID- 9213738 TI - [Formation of observing line direction in the gradient optic systems of superfine medical endoscopes]. PMID- 9213739 TI - [Transesophageal cardiac stimulator Stikar-39 for emergency therapy]. AB - A portable "STICAR-39" apparatus for emergency transesophageal and endocardial pacing is described. The apparatus is the smallest, most light-weighed, and power efficient transesophageal cardiac stimulator in the world. It is effective in emergency medical care, urgent and critical cases, surgery, resuscitation, and intensive therapy. The high cost-efficiency of the apparatus is achieved by using the original scheme of an impulse accumulator-converter of constant power supply voltage to the voltage of stimulating impulses. PMID- 9213740 TI - [Main principles in design and manufacture of devices and units for psychophysiological studies]. PMID- 9213741 TI - [Medical engineering complexes for automated insulin therapy of diabetes]. AB - The paper deals with the automatization of therapy for diabetes, one of the metabolic diseases, which is based on medical engineering complexes to provide an accurate correction of abrupt-onset highly progressive pathological processes and to ensure the maximum physiological adequacy of therapy. The paper also gives results of numerous investigations of medical engineering complexes for automatic insulin therapy and reveals the tends and problems concerned with their development. PMID- 9213743 TI - [Portable medical devices permitted by the RF Ministry of Health for domestic use]. AB - The paper presents information on some portable physiotherapeutical apparatuses used for domestic application. These include topical heating and magnetic therapeutical equipment. PMID- 9213742 TI - [DZR-EK x-ray dosimeter]. AB - Introducing accessory ionization chambers and refusing an operation-mode switch ensure convenience for deal with an alleviated dosimeter and a high accuracy of its readings. The device is designed to check the radiation output of an X-ray radiator and the radiation sensitivity of an image receiver (such as an X-ray image amplifier). PMID- 9213744 TI - [Transcranial electrostimulation: a new approach (experimental and clinical bases and equipment)]. AB - The paper deals with one type of electrostimulation-transcranial electrostimulation (TES). It outlines the physiological mechanisms underlying TES, the clinical features of its application, the equipment performing TES, defines its optimum analgesic mode. PMID- 9213745 TI - Measles eradication: recommendations from a meeting cosponsored by the World Health Organization, the Pan American Health Organization, and CDC. AB - Recent successes in interrupting indigenous transmission of measles virus in the Americas and in the United Kingdom prompted the World Health Organization (WHO), Pan American Health Organization (PAHO), and CDC to convene a meeting in July, 1996 to consider the feasibility of global measles eradication. Presentations at the meeting included an overview of global measles control and elimination efforts; detailed reviews of successful measles elimination efforts in Latin America, the English-speaking Caribbean, Canada, and the United States; surveillance for clinical disease; laboratory tools for antibody detection and virus identification; and other factors that might influence the feasibility of disease eradication. With this background information, meeting organizers asked participants to address five questions: 1) Is global measles eradication feasible? 2) Is measles eradication feasible with current vaccines? 3) What are the appropriate vaccination strategies for measles eradication? 4) How should surveillance for measles be carried out? 5) What role should outbreak control play in the strategy to eliminate measles? Participants agreed that measles eradication is technically feasible with available vaccines and recommended adoption of the goal of global eradication with a target date during 2005-2010, with the proviso that measles eradication efforts should not interfere with poliomyelitis eradication but should build on the successes of the global Poliomyelitis Eradication Initiative. Although existing vaccines are adequate for eradication, vaccination strategies that rely on administration of a single dose of vaccine are not. In the Americas, sustained interruption of indigenous measles virus transmission has been achieved through a three-tiered vaccination strategy that includes a) "catch-up" vaccination of all persons aged 1-14 years, regardless of disease history or vaccination status; b) "keep-up" vaccination of > or = 90% of children in each successive birth cohort at age 12 months and c) "follow-up" campaigns designed to vaccinate all persons within a specific age range whenever the number of susceptible persons in the preschool-aged population approximates the size of a typical birth cohort (in practice, every 3-5 years). In other regions, different strategies may be optimal. Surveillance, a critical component of any strategy to eliminate or eradicate measles, has two functions: to assess the effectiveness of the measles elimination strategy and to detect circulation of measles virus in a population. Systematic surveillance based on clinical diagnosis should be implemented early in any measles elimination program. In countries attempting to eliminate indigenous measles, all isolated cases of measles and at least one case in each chain of transmission should be confirmed by laboratory tests. Specimens for virus isolation (e.g., urine, nasopharyngeal swabs, or blood) should be collected in conjunction with field investigations. Vaccination campaigns generally have not proved to be effective responses to measles outbreaks. Outbreaks should be treated as opportunities to reinforce surveillance and to identify measures to prevent future outbreaks. The major obstacles to measles eradication are not technical but perceptual, political, and financial. Measles is often mistakenly perceived as a mild illness. This misperception, which is particularly prevalent in industrialized countries, can inhibit the development of public and political support for the allocation of resources required for an effective elimination effort. The disease burden imposed by measles should be documented, particularly in industrialized countries, so that this information can be used to educate parents, medical practitioners, public health workers, and political leaders about the benefits of measles eradication. PMID- 9213746 TI - [Molecular markers in genetic mapping of plants]. PMID- 9213747 TI - [DNA vaccination and gene therapy based on transient expression of nucleic acids in human and animal somatic cells]. PMID- 9213748 TI - [Expression of the human apolipoprotein A-I gene transferred in vitro into mammalian cells and in vivo into rat liver]. PMID- 9213749 TI - [Functional status of the p53 gene in murine F9 embryonal teratocarcinoma cells]. PMID- 9213751 TI - [Periodicity of distribution of trinucleotides in DNA sequences, found in the synaptonemal complex of the golden hamster]. PMID- 9213750 TI - [Electron microscopic studies of compactization of circular DNA in model systems. I. Compact structure of complexes of the synthetic peptide trivaline with circular DNA, containing a sequence, causing a stationary bend in DNA]. PMID- 9213752 TI - [Restriction mapping of a highly-repetitive DNA sequence in studying the genetic relationship of lower taxons of animal]. PMID- 9213753 TI - [Protein kinase activity strongly related to bovine tryptophanyl tRNA synthetase]. PMID- 9213754 TI - [Complex formation of ethidium bromide with the single-stranded noncomplementary deoxytetranucleotide 5'-d(ApApGpC)]. PMID- 9213755 TI - [Interaction of mono- and bisbenzimidazole analogs of Hoechst-33258 with DNA]. PMID- 9213756 TI - [Composition of the hydrophobic stem and stability of double-stranded parallel superhelical alpha-helix]. PMID- 9213757 TI - [Interaction of DNA molecules with coordination complexes of divalent platinum. III. Platinum compounds with two pyrimidine ligands]. PMID- 9213758 TI - [Brownian dynamics of bovine serum albumin assessed by a proton relaxation method]. PMID- 9213760 TI - [Structure and conformational dynamics of (dA.dT:dT)6 with parallel oriented thymine chains]. PMID- 9213759 TI - [Structure and conformational dynamics of (dA.dT:dT)6 with antiparallel oriented thymine chains]. PMID- 9213761 TI - [Preparation of a retinoblastoma gene with a mutation in the C-domain and assessment of growth-suppressing activity of its product]. PMID- 9213762 TI - [Isolation of 8-oxoguanine-DNA-glycosylase from Escherichia coli and substrate specificity of of the enzyme]. PMID- 9213763 TI - [Effect of amino acid substitutions in the gp120 V3-loop sequence on serological cross-reactivity between various HIV-1 subtypes]. PMID- 9213764 TI - [Photoaffinity modification of human immunodeficiency virus reverse transcriptase with an analog of deoxyuridine-5'-triphosphate, containing an arylazide group]. PMID- 9213765 TI - [Regularities in utilization of deoxyribonucleoside triphosphates by mutant forms of bacteriophage T7 RNA polymerase]. PMID- 9213766 TI - [Inhibition of the polymerization reaction, catalyzed by human immunodeficiency virus reverse transcriptase using model substrates by derivatives of oligo-1,3 thiazolecarboxamides]. PMID- 9213767 TI - [Design, synthesis, and immunochemical identification of the proposed receptor domain of human insulin]. PMID- 9213769 TI - [Antibiotic and electrochemical properties of some natural ionophoric compounds from streptomycetes]. AB - The systematic position of 8 Streptomyces strains isolated from nature was determined as a result of purposeful search for producers of ionophore compounds; 7 of them belong to different species. Study of the antibiotic activity of these strains and of antibiotics isolated from them as crystals permit us to hypothesize that Streptomyces may be the test organisms most fit for production of hydrophobic ionophores from streptomycetes. Alteration of the ranges of cationic and anionic selectivity by using different electrolytes indicates a high lability of the natural ionophore compounds, which may be due to their physiological function: to take an active part in the exchange processes of all inorganic ions between the cell and environment. This very property may seriously impede the use of ionophores for some practical purposes, e.g., making ion selective electrodes. PMID- 9213768 TI - [Interaction of human immunodeficiency virus reverse transcriptase with oligonucleotide derivatives containing polycyclic aromatic groups at the 5' terminus]. PMID- 9213770 TI - [Essays on modern molecular genetics from lectures for students of the biology department at Moscow State University. Essay 6. Gene therapy and medicine of the XXI century]. PMID- 9213771 TI - [Cloning and expression of the gene for thermostable beta-galactosidase from Thermoanaerobacter ethanolicus in Escherichia coli: purification and properties of the product]. AB - An anaerobic thermophilic bacterium Thermoanaerobacter ethanolicus 39E (Clostridium thermohydrosulfuricum 39E) gene library was constructed in E. coli. Recombinant plasmid (pUT50) containing the thermostable beta-galactosidase was isolated by direct selection of clones for enzyme activity using 5-bromo-4-chloro 3-indolyl-D-galactopyranoside (X-gal) and mapping procedures were carried out. The beta-galactosidase was purified from cell extracts of E. coli. Physicochemical characteristics of the recombinant beta-galactosidase were determined. The enzyme has two optimum pH values: 5.3 and 6.0, the temperature optimum is 75-80 degrees C. The molecular weight of beta-galactosidase was determined by PAG electrophoresis: about 83 kDa. PMID- 9213772 TI - [Prevalence of IS285 and IS100 in Yersinia pestis and Yersinia pseudotuberculosis genomes]. AB - Cell DNAs of various species of Enterobacteriaceae were hybridized with the probes based on IS285 and IS100, mobile genetic elements of Yersinia pestis. These IS elements are found only in the genomes of all tested Y. pestis strains and a number of strains of a related bacterium Y. pseudotuberculosis. Phylogenetic relations between the tested strains and correlation of fingerprints with the geographical origin of the strains were revealed by analysis of the hybridization profiles of Y. pestis and Y. pseudotuberculosis chromosomal DNAs with IS probes. Comparison of the chromosomal IS100 profiles of Y. pestis wild strain and its nonpigmented mutant helped us determine the minimal extension of the genetic rearrangement and detect at least three copies of the IS element in the mutant region. PMID- 9213773 TI - Prevalence of the rickettsial agent of human granulocytic ehrlichiosis in ticks from a hyperendemic focus of Lyme disease. PMID- 9213774 TI - Residual clones in childhood leukemia. PMID- 9213775 TI - Residual clones in childhood leukemia. PMID- 9213776 TI - Residual clones in childhood leukemia. PMID- 9213777 TI - Venous thromboembolism. PMID- 9213778 TI - Venous thromboembolism. PMID- 9213779 TI - Massive pulmonary embolism. PMID- 9213780 TI - Massive pulmonary embolism. PMID- 9213781 TI - Shortage of intravenous multivitamin solution in the United States. PMID- 9213782 TI - Physician-operated networks and antitrust regulations. PMID- 9213783 TI - Care of the woman who has been raped: correction. PMID- 9213784 TI - Physician-assisted suicide and patients with HIV disease. PMID- 9213785 TI - Physician-operated networks and antitrust regulations. PMID- 9213786 TI - [Risk of respiratory insufficiency caused by thoracic rigidity]. AB - Three patients, a man aged 71 and two women aged 47 and 54, were admitted for chronic obstructive pulmonary disease and cardiac failure. All three had thoracic deformities, owing to earlier pneumonectomy with thoracoplasty because of pulmonary tuberculosis, congenital kyphoscoliosis, and infant poliomyelitis respectively. Such patients are at risk of developing chronic respiratory insufficiency because of chronic alveolar hypoventilation: muscle power decreasing with age gradually fails to meet the increased respiratory labour. Often, the respiratory insufficiency is not noticed because the problems are ascribed to secondary chronic obstructive pulmonary disease or cardiac failure. The first sign of imminent respiratory insufficiency is nocturnal carbon dioxide accumulation. Therapy consists of respiratory assistance at night by positive air pressure ventilation via a nose mask. PMID- 9213787 TI - [A new approach to rheumatoid arthritis]. AB - Slow-acting antirheumatic drugs (SAARDs) are usually prescribed in rheumatoid arthritis only when non-steroid anti-inflammatory drugs can no longer suppress the inflammation or when articular damage is radiologically apparent. It was established recently that articular damage occurs in an early phase of RA. This damage is linked to subsequent disability; to prevent it is the purpose of SAARDs. In view of the short-term reduction of arthritis activity and improvement of function as well as the meanwhile established fact that the side effects of SAARDs are not different from those of other antirheumatic agents, SAARDs should be prescribed in an early phase of RA. PMID- 9213789 TI - [Clinical judgment and decision making in clinical practice. A music conductor with epilepsy followed by memory disorders]. AB - A 40-year-old conductor was admitted because of increasing drowsiness and confusion. Two years before admission he had had a first seizure. One year before admission he had a generalized convulsive status epilepticus; the following months he was less able to concentrate. A second status epilepticus was followed by transient weakness of his left arm and a depressed level of consciousness for several weeks. After awakening, he had delusions, and his wife found him demented. In the following months his confusion and drowsiness gradually deteriorated. He had previously had gonorrhoea, an episode of fever and exanthema, and was found to have oligospermia as cause of his infertility. On examination he was disoriented, and he had dysarthria. His left pupil was smaller, but both pupils reacted normally. There was left hemianopia and cerebellar ataxia. CT and MR showed large ventricles and periventricular diffuse lesions in the white matter. CSF examination revealed leucocytosis and increased protein content. Further examination were focussed on serological evidence of syphilis, and finally neurosyphilis was diagnosed. After treatment with penicillin, the patient started to recover. PMID- 9213788 TI - [Clinical application of albumin: a closer look at indications]. AB - Albumin infusions are given far from always on the correct indications, and often there are alternatives that are cheaper and equally suitable. In septic or hypovolaemic shock, crystalline fluids are cheaper and equally efficacious for volume therapy. It is only in sporadic patients with a nephrotic syndrome that colloidal solutions such as albumin are indicated in hypovolaemia. Albumin infusion has no place in the combating of oedema. In decompensated hepatic cirrhosis with ascites, it appears useful to combine paracentesis with albumin infusion, to prevent renal insufficiency and hyponatraemia, but other colloidal fluids are probably equally suitable. Combating hypoalbuminemia as such is not useful in seriously ill patients; it is the underlying disease that should be treated. PMID- 9213790 TI - [Favorable results of early 2nd-stage medication in rheumatoid arthritis]. AB - OBJECTIVE: To compare the effects of treatment with and without 'slow-acting antirheumatic drugs' (SAARDs) for patients with recent-onset rheumatoid arthritis (RA). DESIGN: Open randomized clinical trial. SETTING: Outpatient clinics of six medical centers, the Netherlands. METHOD: The study population consisted of 304 consecutive patients with recently diagnosed RA. A therapeutic strategy with delayed introduction of a SAARD was compared with a strategy comprising early start of SAARD treatment. Primary endpoints were functional disability, pain, joint score, erythrocyte sedimentation rate and progression of radiological abnormalities at 12 months. RESULTS: Four of the five endpoints improved statistically significantly more in the early-SAARD group than in the non-SAARD group; radiological abnormalities progressed at an equal rate in the two groups. Adverse reactions remained at an acceptable level and were the same in the two groups. CONCLUSION: Early introduction of a SAARD was more beneficial than a therapeutic strategy with delayed introduction of a SAARD for patients with recently diagnosed RA. PMID- 9213791 TI - [Resignation on medical grounds: family physicians insufficiently aware of the background]. AB - OBJECTIVE: To inventory the knowledge and the involvement of General Practitioners (GPs) in the procedure "resignation on medical grounds'. Resignation on medical grounds used to be a lawful action in order to prevent further occupational health deterioration. With the advent of new legislation resignation by the employee is financially detrimental as there is no financial compensation for the loss of work, except in case of resignation because of acute medical problems. DESIGN: Exploratory, descriptive. SETTING: North-West and North East Twente region, the Netherlands. METHOD: A short questionnaire was sent to the 160 GPs in this region. RESULTS: Of the notified GPs 72% (n = 116) responded. The familiarity with the concept "resignation on medical grounds' appeared to be great (78%), in contrast to the knowledge of its legal background (22%). The number of times in the preceding 6 months that patients, industrial medical officers and social insurance physicians had approached the GPs with questions regarding resignation on medical grounds was limited; 34% had been consulted. A majority of the GPs considered it their task to make statements on this subject. CONCLUSIONS: There was a time when "resignation on medical grounds' could offer a solution when a working situation was damaging the health of an employee. In the current legislation it is only in cases of acute medical necessity that the resignation is not blame-worthy. The emphasis must nowadays be put on mediation and reintegration. This is the special task of the industrial medical officer. The GP may play a supportive, consultative part. PMID- 9213793 TI - [Familial combined hyperlipidemia]. PMID- 9213792 TI - [Iatrogenic cicatricial endometriosis]. AB - A woman aged 36, who two years previously had undergone a caesarean section, developed a painful swelling of the scar attributed to cicatricial endometriosis. This is a rare disorder brought about by peroperative displacement of normally or ectopically situated uterine mucous membrane to the surgical wound. There, it may proliferate and cause characteristic signs and symptoms that are simple to diagnose: cyclic pain, swellings and sometimes, bleedings. Pharmaceutical, possible hormonal treatment is of limited value, because cessation of the medication is frequently followed by recurrences. Consequently, ample surgical excision in endometriosis is to be preferred; this is to be regarded as a curative therapy. PMID- 9213794 TI - [Chronic central cyanosis in children; always an indication for further diagnosis]. AB - A girl and a boy, both aged 4 years, had displayed a blue discolouration of the skin for several years. In the girl, electrocardiography and roentgenography of the chest revealed no abnormalities; in the boy, the cardiac murmur was attributed to an insignificant ventricular septal defect. Further examinations were performed only when the children developed sleeping problems and decrease of exercise tolerance, respectively. In both, a right-left shunt was discovered caused by a direct communication between the right pulmonary artery and the left atrium, and tetralogy of Fallot, respectively. Both patients' condition improved after operation. Chronic central cyanosis in a child constitutes an indication for consultation of a paediatric cardiologist. PMID- 9213795 TI - [A uniform protocol for liver transplantation in adults in The Netherlands]. AB - Recently a common protocol for liver transplantation in adult patients was established in the two officially registered liver transplantation centres in the Netherlands, Groningen University Hospital and Rotterdam University Hospital. Although the total number of liver transplantations is steadily increasing in the Netherlands, the number of 85 in 1996 is still far below the estimated required number of 150 per year (10 per million of the population per year). If the current trend continues, the number of performed liver transplantations will reach an optimal level in a few years' time, however. The present uniform protocol prevents patients being referred for liver transplantation too early or too late. PMID- 9213796 TI - [Vitiligo]. AB - Vitiligo is a disorder in which, owing to disappearance of melanocytes in the skin, sharply delimited, symmetrically arranged white patches develop. The condition occurs in 1-2% of the population, mostly between the ages of 10 and 30 years, and as often in males as in females. The course is usually progressive with periods of stability. A number of autoimmune diseases and dermatoses coincide with vitiligo. The cause of vitiligo is unknown. Hereditary factors, autoimmunization, neurological disorders and autodestruction have been hypothesized. Repigmentation therapy consists of photo(chemo)therapy use of corticosteroids, and transplantation of pigment cells. For patients in whom this fails and with more than 80% cutaneous involvement, total depigmentation therapy could be considered. PMID- 9213797 TI - [SAPHO syndrome: common denominator for various bone and skin diseases]. AB - The acronym 'SAPHO' stands for synovitis, acne, pustulosis, hyperostosis and osteitis. Three subtypes of SAPHO can be distinguished which have the following features in common: sternoclavicular hyperostosis and sterile inflammatory lesions in both bone and skin. These subtypes are: sternocostoclavicular hyperostosis, chronic recurring multifocal osteomyelitis, and pustular arthro osteitis. Hyperostosis and osteitic lesions may be similar to those seen in malignant bone tumours. Synovitis generally does not lead to bone erosions and one-third of the patients develop sacroiliitis. The SAPHO syndrome and seronegative spondyloarthropathy share some common features (a higher prevalence of the HLA-B27 antigen, occurrence of sacroiliitis and a higher incidence of chronic inflammatory bowel disease and psoriasis). Aetiology and pathogenesis of SAPHO are unknown; prognosis is good. The SAPHO syndrome often runs a protracted course, with intermittent relapses and remissions without resulting serious disability. Treatment is aimed only at symptomatic relief and mainly consists of analgetics and nonsteroidal antiinflammatory drugs. PMID- 9213798 TI - [Preferential posture in infants; serious demands on health care]. AB - OBJECTIVE: To determine the prevalence of preferential posture in infants up to the age of six months; to determine the percentage of referrals for diagnostics and (or) treatment; to gather information about risk factors. SETTING: Infant Health Care (IHC) centres in the Netherlands. DESIGN: Descriptive controlled investigation. METHOD: During September 1995 a total of 7609 infants were examined by 167 IHC doctors for the presence of preferential posture. Data on the physical examination and on the occurrence of risk factors were registered for every child with preferential posture (n = 623) and for a next child of the same age and the same sex without preferential posture. Six months later 468 children with preferential posture were re-examined. RESULTS: The prevalence of preferential posture was 8.2% and was highest in children below 16 weeks of age. The ratio boy:girl was 3:2. Firstborns, premature children and children in breech position at the time of delivery, had a higher risk for preferential posture. The position of the child after the first week of life and the way of feeding proved to be significant risk factors. After six months 32% of the children with preferential posture had been referred for additional diagnostics and, if necessary, treatment. CONCLUSION: Preferential posture is frequently observed and leads to referral, additional diagnostics and (or) treatment in 2.5% of all children up to 6 months of age. PMID- 9213799 TI - [Carbohydrate-deficient transferrin: a new biochemical marker for chronic excessive alcohol consumption]. AB - OBJECTIVE: To assess the usefulness of the biochemical marker 'carbohydrate deficient transferrin' (CDT) in relation to conventional markers for chronic excessive alcohol use. DESIGN: Prospective. SETTING: Addiction clinic Paschalis, Wanssum, the Netherlands. METHOD: Addicts for weaning (n = 125) were questioned at admission about their drinking habits in the last two weeks. Based on the criterion more or less than 60 g alcohol per day, the group was divided into excessive and nonexcessive alcohol users (men: 52 abusers, 51 non-abusers; women: 12 abusers, 10 non-abusers). Mean cell volume (MCV), gamma-glutamyl transpeptidase (gamma GT) and total transferrin were measured in blood collected 2 days after admission, as well as CDT by two methods (CDTect and % CDTriTIA). RESULTS: In men the CDTect test was the most sensitive: sensitivity 82% with specificity 88%. The sensitivity and specificity were 62% and 86% for gamma GT, 50% and 95% for % CDTriTIA, and 34% and 98% for MCV. The combination of a positive CDTect result and a positive gamma GT result gave a predictive value of use of alcohol > 60 g/day of 100%. The results of CDT and gamma GT were also used for a logistic regression model, giving a statistical prediction for excessive alcohol use. The subgroups of women were too small to detect statistical significant differences between tests. CONCLUSION: The CDTect test was more sensitive for the detection of chronic excessive alcohol use than the conventional markers. The combination of gamma GT and CDTect results increased the positive predictive value. PMID- 9213800 TI - [Allergy to Ficus benjamina: at the workplace and at home]. AB - In four patients, two women aged 40 and 42 years and two men aged 49 and 37 years, type I allergy to Ficus benjamina was established. Two patients had been sensitized by contact with these pot plants at their homes. The other two patients were plant growers. F. benjamina is a non-flowering, currently very popular pot plant to be found in both private houses and public buildings. The symptoms comprise itching and swelling of the eyelids, tears, running nose, wheezing and dyspnoea. In one plant grower contact urticaria progressing to dermatitis of the hand was the main symptom. Only one patient had a clear-cut atopy. Both plant growers showed a cross-allergy to other Ficus species. Two patients had a cross-allergy to latex and the associated cluster of tropical fruit (banana, kiwi, avocado, and chestnut). Removal of the ficus plants from the homes and change to another crop or to another occupation completely resolved the complaints of these patients. PMID- 9213801 TI - [Rwanda, a retrospective to aid rendered following the 1994 genocide]. AB - Some 700,000 Tutsi were killed in Rwanda in 1994, and 1.2 million Hutu fled to neighbouring countries. The United Nations and the non-governmental organizations recently issued a document assessing the aid given. Several matters could have been better. Security in the camps, for instance, was a crucial problem and relief workers and armed forces were unprepared for the involuntary cooperation. Also, estimates of numbers of refugees were inadequate and such estimates were manipulated for political purposes. Relief organizations resorted too quickly to import of goods that were available locally, thereby making the aid unnecessarily expensive. The refugees themselves were involved too little in establishing relief priorities. Life of the original population round the camps was disrupted, a fact that received hardly any attention. PMID- 9213802 TI - [Is smoking a cigar or pipe harmful?]. PMID- 9213803 TI - [Cost-effectiveness of influenza vaccination in The Netherlands]. PMID- 9213804 TI - [Sex differences in perceived health]. PMID- 9213805 TI - [There is no evidence of more symptoms with mefloquine than with other drugs in malaria prophylaxis]. PMID- 9213806 TI - [There is no evidence of more symptoms with mefloquine than with other drugs in malaria prophylaxis]. PMID- 9213807 TI - [Cluster headache: misjudged because unknown]. PMID- 9213808 TI - [Farmacotherapeutisch Kompas 1997]. PMID- 9213809 TI - [Iatrogenic collapse; can this be prevented?]. PMID- 9213811 TI - [Floating thromboembolism of the right atrium in pulmonary embolism. Effectiveness of treatment with rTPA]. AB - We describe a case od echocardiographic visualisation of right atrial thrombus in a patient affected by pulmonary embolism. The thrombus had the characteristic aspect of embolus with venous origin. The litic treatment with rTPA, during echocardiographic monitoring, was effective with disappearance of the thrombus and quick improvement of symptoms. PMID- 9213810 TI - [Cardiac myxoma in the elderly. Clinical study]. AB - Cardiac mixoma in the elderly. A clinical study. The clinical features of 13 cardiac myxomas surgically resected are presented. The mean age at presentation was 68 years. Ten were in the left atrium, 5 near the fossa ovalis, 3 at the base of the atrial septum, 1 at the inferior wall and 1 on the anterior leaflet of mitral valve, 3 were in the right atrium, 1 of these was accompanied with a myxoma at the apex of left ventricle. The ECG and the chest X-ray were normal in 9 and in 8 patients, respectively. In 3 patients, the diagnosis was occasionally made by routine 2-dimensional echocardiography. 5 patients presented with fever of unknown origin, arthralgias, weakness, weight loss. None had intracardiac or extracardiac recurrence in the 73 months follow-up. The presentation with constitutional symptoms only like fever of UO, may mimic collagen and neoplastic diseases, vasculitis, lymphomas: the 2-dimensional echocardiography is mandatory to esclude a cardiac myxoma in the elderly. PMID- 9213812 TI - [A quinapril-hydrochlorothiazide combination in the treatment of essential arterial hypertension]. AB - This open, non-controlled study was planned to investigate efficacy and tolerability of fixed combination quinapril 20 mg plus hydrochlorothiazide 12.5 mg for treatment of mild-to-moderate essential hypertension. One hundred and fifty seven patients, with diastolic pressure ranging 105-119 mmHg, were enrolled after two weeks wash-out; during this period patients received placebo. After placebo phase, patients received one tablet/day of fixed combination for six weeks: at the end of this period patients were classified as "responder" (lowering of pressure = or > 10 mmHg) or "non responder"; "non responder" were planned to receive two tablets/day while "responder" maintained the former posology. Patients were reexamined after four weeks: "non responder" were dropped out while "responder" maintained the actual posology. Study was performed in fourteen weeks totally; at the end of this period fourteen patients were classified as "non responder". The fixed combination quinapril 20 mg plus hydrochlorothiazide 12.5 mg can be considered a valid drug for the treatment of mild-to-moderate hypertension. PMID- 9213813 TI - [Classification and seriousness of chronic venous diseases of the lower limbs. A concensus document]. AB - It has been felt for a long time now that there is a need for a simple but complete classification of lower limb venous diseases, and many proposals concerning this matter have appeared in literature. The object of the new classification prepared at Maui, Hawaii, in 1994 is not only that of placing chronic venous diseases of the lower limbs under different profiles (clinical, etiological, anatomical, physiopathological), but also of supplying a numerical score concerning the seriousness of the disease. PMID- 9213814 TI - [Morphologic anomalies of the extracranial internal carotid artery. Our experience]. AB - Cerebro-vascular insufficiency may be caused by morphologic anomalies of the extracranial internal carotid artery (10-15% of symptomatic patients). These alterations are characterized by anomalous elongation which conditions particular attitudes of the carotid: tortuosity, coiling, kinking. In the first case the artery assumes an "S" or "C" shape; in the second the elongation is more emphasized and the artery develops one or more loops; kinking, the most frequent morphologic anomaly, is a sharp angulation of the first part of the internal carotid artery. The etiology of these anomalies seems to be related to congenital causes, that may be unmasked by arterial growing old process. Surgical correction, indicated for symptomatic patients or patients with important hemodynamic alteration, requires rectilinearisation of the internal carotid artery associated with TEA eversion of the same. PMID- 9213815 TI - [Acute intestinal ischemia. Review of the literature and personal experience]. AB - The authors report the experience of their surgical department in the diagnosis and treatment of acute arterial mesenteric ischaemia. Their results are similar to those of the literature. Stress is laid on early diagnosis and interest of mesenteric angiography, laparoscopy and color flow ultrasound. Arterial revascularisation must be the surgical aim but the prognosis of acute mesenteric ischaemia remains very severe. PMID- 9213816 TI - [Hyperhomocysteinemia: a new independent vascular risk factor?]. AB - The positive correlation existing between hyperhomocysteinemia and atherosclerosis has firmly been established through data derived from numerous epidemiologic and experimental observations as well as from intervention trials. Although most of the clinical data have been obtained in relation to coronary heart disease, hyperhomocysteinemia is also observed in patients with cerebral and peripheral arterial occlusive disease or peripheral venous thrombosis. The prevalence of the heterozygous state is today estimated about 1 to 2 percent of the population. Mild and moderate hyperhomocysteinemia have recently been proposed as an additional and independent risk factor for vascular disease. In this review we therefore describe recent findings about the pathogenesis of hyperhomocysteinemia and their implications for optimal drug therapy. PMID- 9213817 TI - [Surgical therapy for prosthetic infections of the thoracic aorta. Conservative approach]. AB - The prosthetic graft infection of the thoracic aorta is a dreaded complication and it is associated with a high mortality rate. There is not substantial agreement in literature about how to manage a vascular graft infection, except for local anti-septic irrigation with a systemic antibiotic therapy. The main point of discussion is if it is mandatory to remove or not the infected thoracic aorta prosthesis: some authors prefer to eliminate all the thoracic aortic prostheses which may be infected, while others propose graft removal only when the sutures lines are involved. In this paper we report our experience on the conservative management of infected thoracic aorta prostheses using a local antiseptic irrigation, a perigraft debridement and leaving the original graft "in situ" when there is evidence of graft damage especially or involvement of the sutures lines. This approach has been performed in three patients: two had an infected aortic arch prosthesis, while one had a descending thoracic aorta prosthesis infection. PMID- 9213818 TI - [Age as a factor in the clinical and prognostic picture and the incidence of myocardial infarct]. AB - The purpose of this review is to determine whether advancing age is an independent predictor of increased mortality after acute myocardial infarction (AMI). Atypical presentations of infarction-related symptoms in the elderly are common, with consequent delay in the diagnosis and treatment. Advancing age is associated with changes in cardiovascular structure and function, that might predispose to adverse outcome of the older infarcted patient, who presents more frequent previous coronary events, ventricular hypertrophy and heart failure. Non cardiac unfavorable data, such as impaired renal function, diabetes and hypertension, are also frequently associated. In elderly patients, several complications of AMI are more common, as external cardiac rupture, cardiogenic shock, heart failure, conduction disturbances. On the contrary, lower values of cardiac enzymes, indicating a lower amount of myocardial necrosis, are observed in older patients. AMI complications are related to the more frequent mortality in elderly patients. The medications proven to reduce mortality, as thrombolytic therapy, aspirin, heparin, beta-blockers, are less frequently employed than in younger patients, despite similarities in a variety of clinical indexes of the extent of myocardial damage. After AMI, coronary angiography is also performed less often in elderly patients; consequently myocardial revascularization with angioplasty or aortocoronary bypass are less employed, despite undoubted therapeutic advantages at all ages. In patients more than 70 years old, AMI affects the female gender more than men; these data involve other particular problems concerning prognosis and therapy. The present benefits, as pointed out by the recent progress in AMI therapy, must be employed in the treatment of older infarcted patients. They go on to suggest that more aggressive management in elderly patients should be evaluated for its potential to reduce mortality. PMID- 9213819 TI - [Isolated cardiac amyloidosis. Presentation of a clinical case]. AB - A 48-year old patient affected by congestive heart failure came to our observation for cardiac arrest due to ventricular fibrillation. After cardiopulmonary resuscitation and defibrillation he underwent complete evaluation. Echo Doppler findings were consistent with restrictive cardiomyopathy. Laboratory findings revealed monoclonal gammopathy and plasma cells dyscrasia. Diagnosis of amyloidosis was then suspected and biopsies of different organs and tissues were performed. Presence of amyloid deposits was found only in myocardial specimens from the right ventricle. Medical treatment with drugs of various classes, administered during hemodynamic invasive monitoring, was uneffective in improving the hemodynamic and clinical status of the patient and he entered in a heart transplantation list. He died six months later, while awaiting for transplantation. Although isolated cardiac amyloidosis is quite rare, we believe that this condition has ever to be kept in mind during differential diagnosis of restrictive cardiomyopathies and we remark that endomyocardial biopsy was mandatory and necessary for certain diagnosis in this case; in addition, the unefficacy of drugs nowadays available for treatment of congestive heart failure in amyloid cardiomyopathy is confirmed. PMID- 9213820 TI - [Clinical course and incidence of post-thrombophlebitic syndrome after profound asymptomatic deep vein thrombosis. Results of a transverse epidemiologic study]. AB - Currently, there are not reliable data on the incidence and prevalence of post phlebitic syndrome (PTS) after an episode of asymptomatic postoperative deep vein thrombosis (DVT). In order to evaluate the epidemiology and the clinical course of PTS in patients who were submitted to hip and knee replacement, we performed a cross sectional study in orthopedic patients with previous asymptomatic postoperative DVT. For reducing potential biases, we used currently accepted and objectively documented criteria to define the clinical manifestations and severity of PTS. RESULTS: 98 of 217 (45.1%) patients, who underwent orthopedic surgery in the previous 2-4 years, have been included in the study. 46 of them (46.9%) had postoperative asymptomatic DVT, confirmed by venograph 23.9% satisfied criteria to be classified as having PTS. When compared with the control group (PTS 3.8%), the difference in the incidence of PTS was statistically significant (p = 0.03). The comparison of the distribution of proximal and distal thrombi in patients with PTS showed that proximal venous involvement constitutes and independent risk factor for developing PST (RR 4). We conclude that: a) about 24% of patients with previous asymptomatic postoperative DVT developed the SPT in the following 2-4 years, b) in asymptomatic DVTs, the localization of thrombi in the proximal venous segment is the most important independent risk factor for the PTS. Therefore, these results can influence the use and the choice of an adequate antithrombotic prophylaxis for reducing the incidence of postoperative proximal DVT. PMID- 9213821 TI - [Up-date on acute ischemia of the limbs]. AB - The authors report their experience in the treatment of acute ischemia of the limbs during the last year. They underline the importance of a right diagnostic and therapeutic treatment particularly in a little hospital, as reported in an algorytm published recently in the literature. They analyze 11 patients, one of them with an embolism of the arm, with a different degree of ischemia. The results are similar to those reported in the literature. PMID- 9213822 TI - [Vascular bench surgery in renal transplant]. AB - The development in the number of patients for renal transplants has not been matched to the kidneys supplied. On this subject the authors think that this chronic deficit could be improved by making use of all the organs by using a series of technical means during bench surgery; which enable optimisation of use of kidneys with vascular abnormalities or those injured upon removal. The authors report their experience of 450 renal transplants operated between January 1981 and December 1985 and of the evolution vascular bench surgical techniques which enable use of a considerable number of kidneys which would otherwise have been discarded. Moreover, it helped the implant, shortened surgery time without prolonging hot ischemia, and did not increase the number of complications. PMID- 9213823 TI - [Profound venous thrombosis and popliteal cyst: problems of differential diagnosis relative to the use of phleboscintigraphy. Description of a clinical case]. AB - The following is the case report of M. G. a 68-year-old male carrier of polcythemia vera, a pathology in which the risk of thrombosis is increased. The patient presented clinically suspected deep venous thrombosis and a phleboscintigraphy confirmed the diagnosis. A real-time B-mode ultrasonography performed later instead demonstrated a popliteal cyst. The case report focuses on diagnostic methods in deep venous thrombosis and particular attention is paid to the role of phleboscintigraphy, of impedance plethysmography and of real time B mode ultrasonography. PMID- 9213824 TI - [Intravascular ultrasonography]. PMID- 9213825 TI - [Electrophysiological analysis of atrioventricular and intraventricular conduction in bi- and tri-fascicular blocks]. AB - We have evaluated, at baseline and during incremental atrial pacing (AP), intracardiac conduction features of 53 patients with electrocardiographic diagnosis of bifascicular or trifascicular block, free from any pharmacological treatment potentially able to affect atrioventricular (AV) conduction system properties. The patients have been subdivided in the following groups: group A (13 patients), with LBBB and a PQ interval > or = 200 msec; group B (14 patients), with RBBB, LAH with a PQ interval > or = 200 msec; group C (8 patients), with LBBB and a PQ < 200 msec; group D (15 patients), with RBBB, LAH and a PQ < 200 msec; group E (3 patients), with RBBB, LPH and a PQ < 200 msec. In group A, 31% presented a long AH interval (> 140 msec), while 85% showed an increased infra-his conduction time (HV > 55 msec). During AP, only 38.5% maintained a 1:1 AV conduction ratio up to 140 bpm, while 30.8% developed an infra-his Mobitz 2 2nd degree AV block. 15.4% an infrahis 2:1 2nd degree AV block, 15.4% an AV nodal Mobitz 2 2nd degree AV block. In group B, 64% and 29% exhibited respectively an AV nodal and an infrahis conduction delay. During AP, 57.1% maintained a 1:1 AV conduction ratio up to 140 bpm, 14.3% developed an AV nodal Mobitz 1 2nd degree AV block, 14.3% an infrahis Mobitz 1 2nd degree AV block, 7.1% an AV nodal 2:1 2nd degree AV block, 7.1% an infrahis Mobitz 2 2nd degree AV block. In group C, no patient manifested a prolonged AH interval, while 50% exhibited a HV > 55 msec. 62.5% maintained a 1:1 AV conduction ratio up to 140 bpm, 25% developed an AV nodal Mobitz 1 2nd degree AV block and 12.5% an infrahis 2:1 2nd degree AV block. In group D, no patient showed an increased AH interval and only 13% presented a HV interval exceeding 55 msec. During AP, 86.7% maintained a 1:1 AV conduction ratio up to 140 bpm, 6.6% developed an AV nodal Mobitz 1 2nd degree AV block, 6.6% an infrahis 2:1 2nd degree AV block. In group E, no patient showed a prolonged AH interval, while 2/3 (66.6%) exhibited an infrahis conduction delay. During AP, 100% developed an infrahis 2:1 2nd degree AV block. Considering all patients with LBBB (groups A+C) and with RBBB+LAH (groups B+D), no differences were found in terms of PQ, PA and AH intervals, even though, concerning patients with a long PQ (group A vs group B), AH interval resulted significantly longer in patients with RBBB+LAH (121.85 +/- 36.4 msec vs 163.29 +/- 55.96 msec, p = 0.031). Infrahis conduction, independently from the measurement adopted (HVI interval: from the beginning of the His to the onset of the ventricular electrogram recorded at the His region; HV2 interval: from the beginning of the His to the onset of the surface QRS), resulted more compromised in patients with LBBB than in patients with RBBB+LAH (HVI: 75.24 +/- 40.23 msec vs 50.79 +/- 25.16 msec, p = 0.011; HV2: 77.24 +/- 38.12 msec vs 53.92 +/- 29.3 msec, p = 0.015). Such a difference became even more significant when comparing the percentage of patients with a prolonged HV interval (average value > 55 msec) in the above mentioned groups: 71.4% in case of LBBB, 20.7% in case of RBBB+LAH (p < 0.001). Regarding intraventricular conduction (IV), no statistically significant differences were found. In patients with RBBB+LAH, IV was not related to infrahis conduction time and PQ interval appeared more related to AH (r = 0.838, p < 0.001) than to HV (PQ-HV1: r = 0.381, p = 0.041, PQ-HV2: r = 0.474, p = 0.009). Conversely, in patients with LBBB infrahis and IV conduction appeared linearly related (HVI-V: r = 0.416, p = 0.06; HV2-V: r = 0.445, p = 0.043). As for PQ interval, it resulted more closely related to infrahis conduction (PQ-HVI: r = 0.626, p = 0.002; PQ-HV2: r = 0.674, p < 0.001), than to AH (r = 0.533, p = 0.013). In conclusion, infrahis conduction resulted more impaired in patients with LBBB. In this group, differently from patients with RBBB+LAH, infrahis conduction seems to affect the degree of IV conduction delay. (ABST PMID- 9213826 TI - [Transeptal access to the mitral valve. Long-term results]. AB - Optimal mitral valve repair or replacement requires an excellent exposure. We used a transeptal approach since 1975 at our Institution to obtain adequate exposure of mitral valve in 135 patients (48 males, 87 females, mean age 47.4 +/- 11.8, range 12-68). A mechanical valve (Bjork = 120; Sorin = 15) was implanted in mitral position. Associated procedures were performed in the 66% of the patients and most of them were tricuspid repair. About half of the patients were at the second or third cardiac operation after a previous closed heart mitral commissurotomy 15.15 +/- 5.6 years before. Exposure was excellent in the 95% of the cases. Hospital mortality was 12.6% and significantly major in patients at redo operation. Three patients with a concomitant aortic valve replacement required a definitive pace-marker implantation. A complete follow-up was possible in all patients who survived at operation. Actuarial survival rate at 10 years in 83% and at 20 years is 70%. Freedom from all events valve related at 10 years is 86% and at 20 years is 74%. None of the patients at echocardiographic follow-up revealed complications related to the transeptal approach to the mitral valve. In conclusion we suggest the use of transeptal approach to the mitral valve in case of redo-operations, concomitant tricuspid repair, small left atrium and in case of mitral valve repair because of the good exposure and the less inherent complications. PMID- 9213827 TI - [Ambulatory phlebectomy. Literature review and personal experience]. AB - Outpatient varicose veins surgery, "Phlebectomie Ambulatoire" (FA) introduced by R. Muller in 1966, is now a widespread technique; modified by many authors with personal tips, FA enables most lower limb varicosities to be treated on an outpatient basis and under local anaesthesia. To achieve good functional results, an accurate preoperative diagnostic examination is mandatory; the authors present a review of the indications of FA and their personal experience. Precision in performing micro-incisions, accurate dissection of the varicosities in the subcutaneous tissue and an adequate postoperative elastic bandage guarantee good aesthetic results. At present the treatment of Saphena magna with FA is debated, but some authors have already reported encouraging results. PMID- 9213828 TI - [Archiving and reporting data in an ambulatory angiologic service]. AB - The authors report their personal experience with the archive of a Laboratory of Vascular Diseases. The most important characteristics requested of an efficacious tool are analyzed and, after a description of the various systems adopted in the past, a new computerized specific archive developed by means of a modern relational database (Microsoft Access) is proposed. PMID- 9213829 TI - [Effectiveness and tolerability of heparan sulfate in the treatment of superficial thrombophlebitis. Controlled clinical study vs sulodexide]. AB - One of the most interesting glycosaminoglycans (GAGs) is heparansulphate, known as the physiological activator of antithrombin III and involved in the maintenance of the antithrombotic potential of uninjured endothelium. The aim of our study was to evaluate the tolerability and effectiveness of heparansulphate with respect to sulodexide, another GAG suitable for the treatment of venous diseases. The study was performed in a open-label, controlled, with parallel and randomized groups, design. Thirty patients (aged 32-72 years) suffering from superficial thrombophlebitis were treated for two weeks with heparansulphate 100 mg t.i.d. or sulodexide 250 LSU b.i.d., both given orally. Some coagulative and fibrinolytic parameters (PT; aPTT; fibrinogen; euglobulin lysis time; t-PA; PAI 1; ATIII; alpha 2-antiplasmin; D-Dimer and platelets count) were assayed at the beginning and at the end of the study. Moreover signs and symptoms of disease (skin trophism; local pain; itch and oedema) were assessed. Heparansulphate and sulodexide were able to reduce signs and symptoms with similar degree and to significantly modify t-PA, alpha 2-antiplasmin and ATIII levels without any difference between treatments. Our issues show that heparansulphate can be useful in superficial thrombophlebitis management. PMID- 9213830 TI - [Relationship between late ventricular potentials and ventricular arrhythmias in patients in chronic dialysis treatment]. AB - BACKGROUND: Arrhythmias are frequent pathology in patients with chronic hemodialysis. We evaluated whether a relatively new technique, signal averaging, could be useful in predicting the development of complex arrhythmias in these patients. METHODS: Thirty-three patients, 18 male and 15 female, subjected to thrice weekly chronic hemodialytic treatment with various dialysis techniques, were studied. Exclusion criteria were the presence of antiarrhythmic and inotropic treatment. The following examinations were carried out in all patients: a Holter dynamic electrocardiography for a duration of 24 hours, begun on the day of dialysis, high resolution ECG pre- and post-dialysis to find out if there were any ventricular late potential (VLP). Four hundred beats were examined in order to obtain a background noise of less than 0.7 microV and a better definition of the signal. The following parameters were considered significant for the presence of VLP: a) filtered QRS duration > 120 msec; b) the root mean square of the signal expressed in the terminal portion of QRS (RMS) < 25 microV) high frequency low amplitude signals duration (HFLA) > 40 msec. A positive value in two of these parameters was considered indicative of the presence of VLP. In addition various pre and post-dialysis indices of dialytic efficiency and a mono and two dimensional echocardiogram with pulsed and color Doppler were carried out. Of the 33 patients studied, ten were excluded because they presented too high a background noise at the high resolution ECG. Of the remaining 23 patients, 13 (56%) presented VLP and nine of these (69%) presented complex arrhythmias. Of the ten patients without VLP, 5 (50%) presented complex arrhythmias. The incidence of arrhythmias was highest during the last two hours of dialysis and in the two hours following it. The patients were then divided into two groups (A and B) according to the ejection fraction (EF) found at the echocardiogram. Group A was composed of 17 patients of whom 8 (47%) presented complex arrhythmias; group B (EF < 45%) was composed of the remaining six patients, who all presented complex arrhythmias. In group A nine patients (53%) out of 17 had LVP, in group B four out of six (66%) had it. All the patients except one presented an increase in the thickness of the ventricular wall and alterations of Doppler transmitral filling rate. Left ventricular hypertrophy was diagnosed in 22 out of the 23 patients. Four patients also had chronic ischaemic heart disease; of these three had LVP. There was no correlation between the presence of LVP and the hemodialytic indices and between the latter and complex arrhythmias. CONCLUSIONS: Our study showed that arrhythmias are more frequent in patients with LVP before dialysis than in those without. The difference was statistically significant (p < 0.006); the incidence of arrhythmias was higher in patients with FE < 45% (p < 0.001). The majority of patients (95%) had left ventricular hypertrophy; only four (17%) had ischaemic heart disease too. It is highly probable that the presence of LVP in our patients can be attributed to hypertension and subsequent left ventricular hypertrophy. As patients with LVP at the end of dialysis had a greater incidence of arrhythmias than those without LVP, we suggest that this method could be useful as a first screening in dialysed patients. PMID- 9213831 TI - [In vitro production of endothelin-1 and prostacyclin by cultured endothelial cells in the presence of polymers]. AB - The aim of this study was to evaluate endothelin-1 and prostacyclin production by human endothelial cells cultured in the presence of polyethylene terephthalate and collagen-coated PET. Cell counting and the assay of endothelin-1 and 6-keto prostaglandin F1 alpha, stable metabolite of prostacyclin, were carried out after 48 hour contact of the cells with the examined materials. Endothelial cell contact with uncoated PET caused a significant reduction in cell number, a significant increase in the production of endothelin-1 and a not significant increase in 6-keto-prostaglandin F1 alpha. The endothelial cell contact with collagen-coated PET caused a highly significant decrease in cell number and a not significant decrease in endothelin-1 and 6-keto-prostaglandin F1 alpha. It was concluded that PET causes both a decrease in cell number and a remarkable increase in endothelin-1. On the contrary, collagen-coated PET determines a decrease in cell number and a slight reduction of endothelin-1 and 6-keto prostaglandin F1 alpha. PMID- 9213832 TI - [Electromechanical dissociation in myocardial infarction. Anatomical-clincal study of 82 cases]. AB - In our experience electromechanical dissociation (EMD) is the most common mechanism of fatal cardiac arrest in patients with acute myocardial infarction (AMI). METHODS: We reviewed retrospectively 82 autopsy cases of AMI in whom the medical record documented EMD as terminal cardiac arrest in order to outline the clinical and pathologic features of different subgroups: 26 cases with external cardiac rupture (CR) were compared with 56 cases without CR. In turn, inside the latter series, 16 cases of sudden EMD were compared with 40 cases of EMD occurring in the terminal phase of cardiac shock. RESULTS: In comparison with those without CR, patients with CR showed at multiple regression analysis less evidence of left ventricular failure (p < 0.05); less extended infarct areas (p < 0.01); more frequent sudden EMD (p < 0.05). Most patients with CR had massive pericardial effusion; cardiac rhythm at the onset of EMD was seldom slow in those cases. In the group without CR no discriminant characteristics were found in cases of sudden EMD vs cases preceded by cardiac shock. CONCLUSIONS: In case of CR EMD occurs in less extensively damaged hearts and is generally sudden; in AMI without CR EMD may affect patients with severe depression of pump performance, but not necessarily in shock. EMD after an AMI may result from several factors: cardiac tamponade is prevalent in the presence of CR; in cases without CR our data don't permit to conjecture a distinct pathogenesis for sudden EMD in comparison with cases preceded by shock. PMID- 9213833 TI - [Mitral valve prolapse associated with the aortic bicuspid valve. Discription of a clinical case]. AB - In the literature, the mitral valve prolapse and bicuspid aorta have been widely discussed as isolated cases or in association with other congenital heart pathologies or systematic illnesses. Nevertheless, they have not been documented contemporarily in the same clinical case. The following case describes a healthy, young, asymptomatic athlete, who has a double valvular heart failure. The defect is occasionally evident during transthoracic echocardiographic examination. The role of echocardiography is stressed taking into consideration the natural lineage and unfavourable reciprocal effect on cardiac hemodynamics, omitting relative implications of familial pathologies. This method is suggested as the means of suitable evaluation for athletes. In fact, this is the best technique to reveral the most precocious modification of cardiac hemodynamic. Consequently, echocardiography allows us to guide and monitor the most appropriate therapy. PMID- 9213834 TI - [Controlled clinical study of doxophylline versus aminophylline in the treatment of acute cardiorespiratory insufficiency syndromes]. AB - Twenty patients of both sexes, hospitalized because they were suffering from acute cardiorespiratory syndromes, have been studied in a clinical blind test on unbalanced groups of patients. This research aims to evaluate the therapeutic efficiency and intravenous tolerance of doxophylline. Patients were observed for 24 hours. Signs and symptoms were recorded on a semi-quantitative scale, and the clinical situation, haemogasanalysis, blood pressure, cardiac frequency, and other analytic controls were performed in order to verify the systemic tolerability. We can conclude, in agreement with the literature, that the group treated with doxophylline showed a better reduction in cardiac frequency, while maintaining the same effects on the respiratory apparatus. PMID- 9213835 TI - [Weaning in cardiopulmonary bypass patients with compromised cardiac function. Comparison of enoximone and dopamine]. AB - OBJECTIVE: Evaluate the effects of enoximone and dopamine in patients with impaired left ventricular function after cardiopulmonary bypass (CPB). DESIGN: Prospective study on a consecutive series of patients subdivided into two groups: enoximone (Group E) and dopamine (Group D). SETTING: Policlinico Umberto I, University La Sapienza of Rome. PATIENTS AND METHODS: Thirty patients undergoing elective myocardial revascularization. Before weaning from CPB the patients received inotropic drugs as follows: Group E: enoximone: bolus: 1 mg/kg in 10 min, and continuous infusion of 5 mcg/kg/min; Gruppo D: dopamine: continuous infusion of 5 mcg/kg/min. Hemodynamic measurements were made using a Swan-Ganz catheter inserted before the induction of anaesthesia. RESULTS: Enoximone has proved to be effective in decreasing pre-load and after-load of both right and left ventricle by a positive lusitropic effect and a reduction of systolic stress, thereby increasing the cardiac index. In group D patients maintenance of cardiac output has been demonstrated to be dependent on a chronotropic effect. As a consequence in group D the increase in rate-pressure product has reached potentially dangerous values, reflecting a marked increase in myocardial oxygen consumption. On the contrary in Group E the increase in rate-pressure product has been much more limited. Finally both drugs have proven effective, since all patients have been easily weaned from CPB. CONCLUSIONS: Enoximone is a useful and easily-handled drug to facilitate weaning from CPB of patients with preoperative impaired ventricular function. PMID- 9213836 TI - [The effectiveness of enoximone in patients with serious left ventricular dysfunction submitted for aorto-coronary bypass]. AB - BACKGROUND: The purpose of this study was to investigate whether the combined positive inotropic and vasodilating properties of enoximone have a short-term benefit when used in patients who underwent open heart surgery. METHODS: From 7/1994 to 1/1995 twenty-six patients with severe myocardial dysfunction (ejection fraction < 35%) were enrolled into a prospective trial before undergoing coronary artery bypass graft. They were randomly selected into two study groups: the first treated with enoximone (group E) and the other one with dopamine (group D). Anaesthesia was the same for both groups using high-dose fentanyl. Buckberg cardioplegia was used. All patients were followed by: conventional monitoring, Swan-Ganz catheter and transesophageal echocardiography. measurements (hemodynamic parameters, end-systolic and diastolic area and left ventricular wall motion) were recorded: after induction of anesthesia, after loading-dose and an intensive care unit. Enoximone- and dopamine infusions were started during weaning from cardiopulmonary bypass and tailored to hemodynamic parameters (cardiac index > 2.8 l/min, wedge pressure < 16 mmHg, mixed venous blood saturation > 65%). Major events were defined as: endotracheal intubation > 36 h, using intraortic balloon pump or centrifugal pump, intensive care timer > 48 h, in hospital cardiac death. Prices, were established by DRG-tables (diagnosis related groups). Statistical analysis were performed by X and "t" Student tests. RESULTS: Cardiac index increased more significantly in group E (CI 1.9-->3.9 vs 2.3-->3.3; p 0.05) thanks to a higher reduction of vascular systemic (SVRI 2889- >1447 vs 2536 -->1565; p 0.005) and pulmonary resistances (PVRI 271-->193 vs 288- >218; p 0.05). Fewer major cumulative events and intensive care costs were observed in group E rather than group D. CONCLUSIONS: Enoximone administer immediately after open heart surgery had more beneficial hemodynamic and clinical effects than dopamine in patients with severe left ventricular dysfunction. PMID- 9213837 TI - [Diastolic dysfunction in cardiac surgery intensive care. Study methods, changes and prognosis]. AB - The natural history of patients with coronary artery disease and diastolic dysfunction who underwent coronary artery bypass grafting (CABG) is not well known. The aims of our study were to evaluate the incidence of diastolic dysfunction, its evolution after CABG and its possible correlation with adverse in-ICU prognosis. We studied 88 consecutive patients scheduled for CABG with not severely depressed left ventricular function (ejection fraction > 35%) and multivessels disease. Buckberg cardioplegia was used for myocardial protection. Diastolic function was investigated by recording mitral and venous pulmonary flow by transesophageal Doppler echocardiography (TEE). TEE examination was performed in operative room pre and post-bypass, at ICU arrival and after three months. Diastolic dysfunction was defined as mild, moderate and severe. Adverse in ICU events were defined as: use of inotropic drugs or ventricular mechanical support, an ICU stay > 24 hours, perioperative myocardial infarction and death. The study group was compared with a control group. T-Student test was used; a p < 0.05 was considered significant. A reduced diastolic function was present in 77% of patients at baseline examination. Diastolic dysfunction did not worsen significantly after hypothermic cardiac arrest and reperfusion. It persisted during ICU stay and normalized after three months from CABG in the majority of patients (85%). Diastolic failure was not associated with an adverse ICU prognosis (adverse events: 18 versus 13%; p = ns). PMID- 9213838 TI - [Memory for postoperative pain six months after discharge from the hospital]. AB - OBJECTIVE: To investigate memory for postoperative pain in Intensive Care Unit (ICU) admitted patients after hospital discharge. DESIGN: Prospective study by direct interviews. METHODS: Six months after hospital discharge, we interviewed adult, postoperative, co-operative patients consecutively admitted to ICU for more than 24 hrs, resident near the hospital, who gave informed consent. We investigated intensity of postoperative pain and recollections of critical care reported by patients. The following data were collected from medical records: type and duration of surgical intervention, type of anaesthesia and fentanyl dose, severity of illness at ICU admission, ICU and hospital (after ICU) length of stay and postoperative administration of morphine. RESULTS: Of 130 patients interviewed, 82 (63%) reported no pain, 27 (21%) low and 21 (16%) more than low pain. Among these 3 groups of patients, there was no statistically significant difference in all the variables collected from medical records. Patients who remembered more than low pain recorded emotional distress more frequently (p < 0.001) and physical discomfort less frequently (p < 0.01) than patients whose pain was absent or low. CONCLUSIONS: Most patients report that postoperative pain, during their ICU stay, was absent or low. Emotional distress seems to be related to memory for postoperative pain. PMID- 9213839 TI - [Anesthesia with sevoflurane vs propofol in elective extracavity surgery]. AB - OBJECTIVE: To compare the cardiovascular effects and recovery characteristics of sevoflurane and propofol anesthesia in 80 ASA I and II patients undergoing elective extracavity surgery expected to last at least one hour. DESIGN: A prospective randomized clinical trial. METHODS: After meperidine and atropine premedication, the patients were randomly allocated into two groups: in the sevoflurane group thiopentone was administered for induction of anesthesia and sevoflurane for maintenance; the propofol group received propofol either for induction of anesthesia or maintenance. All patients received N2O, vecuronium, artificial ventilation and fentanyl as needed. Vital parameters were monitored during anesthesia and two hours later. Recovery times were recorded after anesthesia. Statistical analysis was performed with SAS (Statistical Analysis System). RESULTS: In the sevoflurane group, heart rate and diastolic pressure were slightly higher than in the propofol group. Recovery time was faster after sevoflurane anesthesia. PMID- 9213840 TI - [Enoximone in weaning from mechanical circulation support in pediatric patients]. AB - OBJECTIVE: To assess and record the response to continuous infusion of the phosphodiesterase inhibitor enoximone during weaning from mechanical circulatory support (MCS) and to verify the possibility of success with this indication in pediatric patients. DESIGN: Retrospective study. SETTING: Pediatric cardiac surgery intensive care unit. PATIENTS: Two pediatric patients operated for complex congenital heart disease with low cardiac output syndrome in the immediate postoperative period, evolved in cardiocirculatory arrest despite massive inotropic pharmacological support, and then assisted by mechanical circulatory support. INTERVENTIONS: Weaning from mechanical circulatory support with continuous infusion of enoximone, only in one case preceded by a loading dose and associated with catecholamine infusion. MEASUREMENTS AND MAIN RESULTS: During weaning hemodynamic parameters (LAP, CVP, MAP, HR), SvO2, diuresis, rectal and cutaneous temperatures were assessed and recorded. A serial echocardiographic assessment of left ventricular systolic and diastolic diameters and ejection fraction (EF%) has also been performed every 12 hours. Weaning from MCS using enoximone as inotropic support was possible in both cases. CONCLUSIONS: Enoximone proved to be useful in weaning from MCS in two pediatric patients, despite the difficulty to assess its effect in one of the two cases in which enoximone was used together with high dosages of other inotropic drugs. These initial positive results urge us to further investigate applications of this drug in pediatric patients. PMID- 9213841 TI - Computerized Clinical Simulation Testing (CST(r): recruitment of pilot study. PMID- 9213842 TI - Total ear replantation. AB - Since the first report of successful microsurgical ear replantation in 1980, there have been 12 other cases reported in the English literature. As the number of trained microsurgeons increases, the opportunity to treat the amputated ear with microsurgical techniques should become more common. The reported cases have involved a variety of different mechanisms of injury and methods of treatment. There have been three techniques used to revascularize the amputated ear successfully: primary vascular repair, vein grafting, and use of the superficial temporal vessels as a pedicled vascular leash. Through our own experience and a review of the literature, we have been able to identify certain clinical characteristics that help dictate which technique to use. We report four cases of successful ear replantation, review the various techniques that have been used successfully, and provide treatment recommendations for future consideration. PMID- 9213843 TI - [Pancreatitis during treatment of leishmaniasis with n-methylglucamine antimoniate in a subject infected with HIV]. AB - In this study the authors report the case of HIV patients with visceral leishmaniasis. He presented a pancreatitis with evident clinical signs and high increase of amilasis (9876 U/L) the twelfth day of treatment with meglumine antimoniate. The interruption of therapy was followed by a rapid disappearance of signs and symptoms and a normalization of amilasis. In accordance with the most recent studies, it is expedient, for every patients treated with antimonial drugs, to undergo an accurate amilasis monitoring. PMID- 9213845 TI - [The use of BCG for the prevention of recurrence of superficial bladder tumors: 10 year's experience]. AB - The aim of this study was evaluate the usefulness of intravescical administration of Bacillus of Calmette Guerin (BCG) in patients with superficial bladder cancer. The BCG, an immunostimulating agent, has been introduced in routine clinical use; according to the international literature, it is reported to offer better result than the above-mentioned drugs. A retrospective analysis was made of 210 patients who had undergone a complete TUR for superficial bladder tumor. The irritative symptoms and systemic effects are acceptables considering the benefit of the BCG. The results are of great interest and encourage us to continue this study to confirm therapeutic strength. PMID- 9213846 TI - [The utility of postprandial C-peptide evaluation in type 2 diabetes]. AB - The secondary drug failure is a well known phenomenon in the development of type 2 diabetes mellitus, but an exact definition of this situation is still lacking. The aim of this research was to evaluate the beta-cell reserve in non obese diabetic patients in relation to the metabolic control and the duration of disease. The main aim was to identify values of postprandial plasma C-peptide that can characterize the patients requiring insulin treatment. A daily profile was performed in 135 non obese diabetic patients, within 20% of their ideal body weight. The mean diurnal values (m) and the mean post-prandial increases (delta mpp) of plasma glucose (G), insulin (IRI) and C-peptide (CP) were assessed. Fourty-four patients showed good (NwD-GC: G-m = 138 +/- 3.2 mg/dl) and 91 poor metabolic control (NwD-SF: G-m = 210 +/- 4.8 mg/dl), according to the G-m lower or higher than 150 mg/dl. Beta-cell reserve (CP-delta mpp: 0.70 +/- 0.03 vs 1.39 +/- 0.04 ng/ml) and C-peptide/insulin molar ratio (CP/IRA-delta mpp: 2.36 +/- 0.06 vs 2.80 +/- 0.06) were significantly lower (p < 0.001) in NwD-SF than in NwD GC. NwD-GC and NwD-SF were respectively divided into three subgroups, according to the duration of disease. A progressive reduction of CP-delta mpp and an increase in SF prevalence, from the first to the third decade of diabetes duration, was observed. In both NwD-Gc and NwD-SF the duration of disease inversely correlated with CP-delta mpp (NwD-GC: y = 1.59-0.019X, p < 0.001; NwD SF: y = 1.01-0.023X, p < 0.001). The analysis of the two regression lines showed that patients with CP-delta values lower than 1.0 ng/ml require insulin treatment. In conclusion the duration of diabetes and the progressive reduction of beta-cell reserve represent the major pathogenetic factors in secondary failure. PMID- 9213844 TI - [Variations in the origin of the artery of Adamkiewicz]. AB - Variations of the origin of the medullary artery. The purpose of this study was to define the origin of the medullary artery through medullary angiography in order to prevent paraplegia during surgery of the thoraco-abdominal aorta. Twenty eight patients, candidate to thoracoabdominal aorta operations, have been studied for the study of the origin of the medullary artery and its eventual reimplantation during surgery. The artery has been localized in 24 of the 28 patients. In 22 cases (78.6%) it originated from an intercostal artery between D8 and D12, while in 6 cases (21.4%) it originated from a lumbar artery between L1 and L2. In 23 (82.1%) cases it originated from the left side, while only in 5 (17.9) from the right side. Although medullary artery originates more frequently at the thoracic level, particularly from the left D9 and D10, its variability is important, and its localization before thoraco-abdominal aortic surgery is often desirable. PMID- 9213848 TI - [Severe infections caused by group A beta-hemolytic streptococcus: report of the latest 3 cases at the La Fe Hospital of Valencia]. PMID- 9213847 TI - [Clinical features and ultrastructure of primary ciliary dyskinesia and Young syndrome]. AB - A study was conducted of the clinical manifestations and ultrastructure in a series of seven patients with repeated pulmonary infections and infertility (3 cases with primary ciliary dyskinesia [PCD] and 4 with the young syndrome [YS]). Clinical and functional respiratory changes were more marked among cases of PCD. The seminogram showed azoospermia in cases with YS and hypospermia with marked hypomotility in cases with PCD. A nasal mucosa biopsy specimen was obtained from all patients to perform a transmission electron microscopy (EM) investigation. Patients with YS did not have pathognomonic ultrastructural changes, whereas patients with PCD had a large number of ciliary abnormalities (23.3% +/- 1.5%); 14% +/- 7% of them corresponded to nonspecific ciliary changes and the remaining abnormalities to congenital ciliary changes (ciliary disorientation in three cases, defective radial spokes in one case and microtubule transposition in one case). EM is a useful technique which is recommended for the differential diagnosis in this group of patients. PMID- 9213849 TI - [Drug adverse reactions: a practical protocol]. PMID- 9213850 TI - [Basic cardiopulmonary resuscitation in the mountains and other isolated situations]. PMID- 9213852 TI - [Male with sensory-motor polyneuropathy]. PMID- 9213851 TI - [Abdominal pain, arterial hypertension, joint pain, and rapidly progressive kidney failure]. PMID- 9213853 TI - [Headache and decreased visual acuteness of sudden onset]. PMID- 9213854 TI - [Anterior extramuscular neck mass]. PMID- 9213855 TI - [Male with fever and behavioral disorders]. PMID- 9213856 TI - [Excipients and sources of therapy information]. PMID- 9213858 TI - ["Anti-obesity master formulations" and "miraculous products" (reply)]. PMID- 9213857 TI - [Do doctors know the cost of the treatments they prescribe, or is it a matter of luck? (reply)]. PMID- 9213859 TI - [Apolipoprotein E and Alzheimer's disease]. PMID- 9213860 TI - [Angiotensin-converting enzyme: structure, function, and clinical usefulness]. PMID- 9213862 TI - [Angiotensin-converting enzyme in pulmonary thromboembolism as a marker of vascular lesion]. AB - In pulmonary thromboembolism (PTE) metabolic changes can occur, as in serum levels of angiotensin converting enzyme (ACE); therefore, the measurement of serum levels of this enzyme might be useful for PTE evaluation. The objective of this study was to determine changes in serum ACE in patients with PTE and their possible variations after therapy. MATERIALS AND THERAPY: Thirty-one patients with PTE were studied (15 males and 16 females). The mean age was 56 +/- 16 years (range 19-82 years). The patients were included if they had: a) a suspect diagnosis of PTE; b) confirmed by high probability V/Q gammagraphy, or c) data of intermediate or low probability with positive lower limb phlebography. Patients with associated diseases, under therapy with ACE inhibitors or lost during the six months of therapy were excluded from the study. STUDY DESIGN: Patients were evaluated in the acute phase of PTE and after therapy (six months later). Biochemical, gasometric, spirometric, V/Q gammagraphy and ACE parameters were investigated. All of them were compared with reference values and those obtained in both phases of PTE. STATISTICS: The Student "t" test was used for independent parameters with the Bonferroni correction for multiple contrast and a = 0.05. The Pearson regression analysis was used for correlations. The ROC curve was used to study its usefulness at diagnosis. RESULTS: ACE decreased by a 20.5% in the acute phase (31.69% +/- 10.34 mumol/min/l) and turned to normal values (39.91 +/- 9.75 mumol/min/l) at post-therapy phase. This decrease was related with acute hypoxia and a decrease in the lung vascular bed. The analysis of the ROC curve showed an area of 0.69 and a negative predictive value of 91.67% for ACE values higher than 46 mumol/min/l. CONCLUSIONS: ACE activity is a marker for pulmonary pathology which might be indicative of injury and/or decrease of the lung vascular bed; its measurement can be useful in the clinical follow-up of PTE. A return to normal values should be interpreted as improvement in the perfusion of the lung vascular bed. PMID- 9213861 TI - [Comparative study of misoprostol and nifedipine in the treatment of Raynaud's phenomenon secondary to systemic diseases. Hemodynamic assessment with Doppler duplex]. AB - OBJECTIVE: To evaluate the mid-term efficiency and therapeutic safety at a mid term of the orally administered misoprostol, a synthetic PGE1, analogue, compared with nifedipine for the treatment of RP secondary to autoimmune systemic diseases. METHODS: A double blind, crossover study was designed. Patients were randomly distributed to receive either retard nifedipine (20 mg/12 hourly) and misoprostol (200 micrograms/12 hourly) in 10-day periods (washing period with placebo for 10 days). At the end of each period a clinical assessment was obtained on the frequency and severity of symptoms as well as on secondary drug reactions. Simultaneously, blood flow changes in radial artery were Doppler duplex investigated (pulsatility index, resistance index). RESULTS: Twenty patients were studied (15 women and 5 men). The mean basal daily frequency of attacks was 4.8 +/- 2.0 compared with 2.4 +/- 1.4 with nifedipine (p < 0.001) and 2.6 +/- 1.2 with misoprostol (p < 0.001). The mean basal severity of attacks, according to a pre-established scale decreased from 3.7 +/- 0.6 to 1.9 +/- 0.9 with nifedipine (p < 0.001) and to 2.0 +/- 1.0 with misoprostol (p < 0.001). The mean basal value of blood flow in radial artery was 24.9 +/- 14.4 ml/min; with nifedipine it increased to 43.0 +/- 19.2 ml/min (p < 0.001) and with misoprostol to 46.9 +/- 19.2 ml/min (p < 0.001). Five patients (25%) had secondary effects with nifedipine and three (15%) with misoprostol; in no case had therapy to be discontinued. CONCLUSIONS: Misoprostol was similar to nifedipine for the treatment of Raynaud phenomenon secondary to systemic diseases and can be a therapeutic alternative for these patients. PMID- 9213863 TI - [Clenbuterol poisoning. Clinical and analytical data on an outbreak in Mostoles, Madrid]. AB - OBJECTIVE: To report the clinical manifestations and analytical findings in an epidemic outbreak of acute food poisoning with clenbuterol. MATERIALS AND METHODS: The clinical manifestations, physical examination findings and results of complementary tests are reported of fifteen patients performed by veal liver contaminated with clenbuterol. The clinical course of patients at 72 hours is reported. A quantitative measurement of clenbuterol in urine specimens from patients and in a veal liver specimen was performed by high pressure liquid chromatography (HPLC). RESULTS: The male/female distribution of patients was 7/8 respectively, with age ranging from 6 to 44 years. Symptoms appeared after 30 minutes to 2 hours of having ingested veal liver in 93% of cases. Patients presented at the Emergency Department with tremors, palpitations, anxiety, malaise, nausea, and pruritus as the most common complaints. On physical examination tachycardia was noted in 100% of cases. The analytical data included mild hypokaliemia (66%) and leukocytosis (28%). Only one patient required hospital admission on account of an hypertensive crisis. After 72 hours, 67% of patients were asymptomatic. The remaining patients had mild symptoms which included headache, myalgia, asthenia and anorexia. Serum potassium values returned to normality (p < 0.05). Urine measurements of clenbuterol were positive for all analyzed cases (50 +/- 42 ng/ml). The concentration of clenbuterol in a veal liver sample was 500 ppb. CONCLUSION: Clenbuterol poisoning should be suspected when symptoms of adrenergic hyperstimulation occur after the ingestion of meal, usually liver. Common symptoms include tachycardia and mild hypopotasemia. Diagnosis is confirmed by quantitative measurement of clenbuterol in urine. Most patients improve spontaneously shortly afterwards. PMID- 9213864 TI - [Acute pancreatitis: clinical and therapeutic study with somatostatin]. AB - A clinical and therapeutical study of 47 patients with the diagnosis of acute pancreatitis is reported. According to Ranson's criteria patients were initially classified as suffering from mild (28) and severe (18) acute pancreatitis. Twenty eight, 11, 7 and 1 patients had biliary, alcoholic, idiopathic and neoplastic causes, respectively, of their conditions. The classification of episodes was made on the basis of clinical manifestations, biologic investigations, and imaging diagnosis, and is shown in the corresponding tables. The therapeutic profile was a randomized double-blind study: perfused somatostatin (SS) versus placebo (P) (physiologic saline 0.9%). The administration of somatostatin in perfusion (250 mcg/h/48 hours) did not improve significantly the parameters used to score the severity, although the mortality rate decreased significantly (p < 0.05) in the group of patients with the severe form of the disease. PMID- 9213865 TI - [The Duke Health Profile (DUKE)]. AB - The Duke Health Profile is a 17-item generic questionnaire instrument designed to measure adult self-reported functional health status quantitatively during a one week time window. It is appropriate for both patient and non-patient adult populations. It can be self-administered by the individual respondent or administered by another person. The administration time is less than five minutes. It is crucial that each question is answered. There are 11 scales. Six scales (i.e., physical health, mental health, social health, general health, perceived health, self-esteem) measure function, with high scores indicating better health. Five scales (i.e., anxiety, depression, anxiety-depression, pain disability) measure dysfunction, with high scores indicating greater dysfunction. Most extensive use has been in family practice patients with the broadest spectrum of diagnoses, but it has also been used in patient populations with specific diagnoses such as insulin-dependent diabetes mellitus, endstage renal disease, ischemic disease, and impotence. Both internal consistency (Cronbach's alpha) and temporal stability (test-retest) testing have supported reliability of the DUKE. Validity has been supported for the DUKE scales by: (a) comparison of the DUKE scores with scores of other health measures for the same patients, (b) comparison of DUKE scores between patient groups having different clinical diagnostic profiles and severity of illness, (c) prediction of health-related outcomes by DUKE scores. Convergent and discriminant validity have been shown when comparing with other instruments. PMID- 9213867 TI - [Implications of the federal government's austerity package in the field of intramural medical rehabilitation]. AB - The austerity legislation passed September 13, 1996 as well as the additional austerity measures pending for the field of rehabilitation in the framework of legislation aimed at refocusing self-government and self-responsibility in the statutory health insurance scheme (1. und 2. Gesetz zur Neuordnung der Selbstverwaltung und Eigenverantwortung in der gesetzlichen Krankenversicherung 1st and 2nd GKV-NOG) are going to jeopardize insurees' provision with medical rehabilitation generally. For 1997 alone, economies totalling some 3.2 billion DM have been legislated. By drastically increasing the additional payments to be made, the insured are to an intolerable extent being subjected to contributing towards the cost of their medical treatment. What is more, the 2nd GKV-NOG is to enable health insurance funds to totally exclude rehabilitation benefits from the standard catalog of benefits to be provided. Along with these spending limitations in the social insurance schemes, additional burdens have been imposed on the insured in gainful employment under regulations in the field of labour law effective as of October 1, 1996 applicable in case of participation in an in patient rehabilitation measure. Thus, the employer is entitled to make deductions with regard to an employee's claim to continued remuneration during sick leave as well as part of his annual paid recreational leave for the duration of the inpatient rehabilitation measure. The population most adversely affected by this policy are those with chronic illness and disability. PMID- 9213866 TI - [Consequences of the need for cost control in rehabilitation]. AB - Beginning 1997, considerable cost-cutting constraints have become effective which are entirely founded by fiscal reasons. The resulting restrictions will not remain without consequences for care and supply of patients with chronic diseases and handicaps as well as for the entire rehabilitation system itself. The expected cuts are so serious that they will also affect further rehabilitation prospects and objectives. The extent of the austerity measures-especially the extent of budget capping-is running counter to the factual development of needs. PMID- 9213868 TI - [Considerations for the continuing development of vocational rehabilitation from the viewpoint of the Federal Institute for Employment]. AB - The law on enhancing growth and employment (Wachstums- und Beschaftigungsforderungsgesetz-WFG) of September 25, 1996 has entailed fundamental changes in the law governing vocational rehabilitation under the employment promotion act (Arbeitsforderungsgesetz-AFG). While a legal entitlement existed so far, vocational rehabilitation benefits, as of January 1, 1997, on principle are granted as discretionary benefits. In the employment promotion reform bill (Arbeitsforderungs-Reformgesetz-AFRG) awaiting passage in 1997, the former legal entitlement is reestablished in those cases where special vocational rehabilitation benefits and services are needed due to the type or severity of disability, or for safeguarding rehabilitation success. The WFG is to bring about a 500 million DM reduction in vocational rehabilitation spending by the federal employment service Bundesanstalt fur Arbeit (BA). This targetted economy also is adhered to notwithstanding the said major expansion of legal entitlement. Already before passage of the WFG, the BA's governing board had set up a working group to look into the issue of developing vocational rehabilitation further. Subjects dealt with had included cooperation among rehabilitation agencies; participant structure in vocational retraining centre programmes; programmes offered; quality assurance; and cost structures in retraining centres. In the wake of the working group's deliberations, the board passed revised provisions for programme participation in vocational retraining centres, and decided on December 18, 1996 that, as of April 1, 1998 a new settlement procedure should be introduced with the vocational rehabilitation facilities, with specific programme-related prices on the basis of agreed quality standards. PMID- 9213869 TI - [The future of vocational rehabilitation--vocational rehabilitation for the future. An explanation with the federal government's austerity measures]. AB - Under the law on enhancing growth and employment (Wachstums- und Beschaftigungsforderungsgesetz-WFG), the legal entitlement to vocational rehabilitation hitherto stipulated in the employment promotion act (Arbeitsforderungsgesetz-AFG), has been restricted to a narrowly defined population; and pension insurance scheme spending in the entire rehabilitation field been "capped" to the 1993 level minus some 600 million DM. Moreover, the transitional allowance applicable for pension insurance rehabilitees will be lowered. In addition, economies amounting to some 500 million DM have been imposed on the federal employment service Bundesanstalt fur Arbeit. These measures are placing persons with disability at a disadvantage, accept exclusion of entire groups of disabled persons, and endanger the very existence of numerous rehabilitation facilities previously established with significant amounts of public funding. A blind eye is being turned on the high level of demand for qualification measures, on the overall economic benefits of rehabilitation measures, and on the fact that measures of this kind are disabled persons' only chance to hold their own in the face of labour market competitiveness. Also, poor awareness seems to exist of the fact that, in the longer run, meaningful contributions to greater economy will more likely be generated by structural adjustment, increased effectiveness, and greater flexibility. The future of vocational rehabilitation is being placed at risk--notwithstanding that vocational rehabilitation for the future is imperative. PMID- 9213870 TI - [Geriatric rehabilitation needs in Cologne: the concept of "integrated geriatric rehabilitation"]. AB - Given a growing number of elderly people and the related consequences for the health care system, such as increasing numbers of persons with chronic diseases or in need of long-term nursing care, the present rehabilitation system must be extended and new services be introduced. Hospitals in Cologne report that an average 48% of their geriatric patients are in need of rehabilitation measures. Taking into account the patients' willingness and capacity to undergo rehabilitation treatment, the figure reduces to 30%. Based on the number of geriatric patients treated in hospital each year, rehabilitation facilities for 15416 geriatric patients a year have to be provided. About two thirds of the patients require continued hospital treatment after the acute phase. Obviously, these treatments are not only performed in the geriatric departments of the hospitals, as only 161 geriatric beds are available in Cologne. One third of the patients could be taken care of on a partial hospitalization or outpatient basis after the acute phase. Thus, limiting a patient's length of stay in hospital could have a cost-containment effect. As partial hospitalization and outpatient rehabilitation facilities are lacking, the Cologne "Geriatrics" study group has developed a concept of "integrated geriatric rehabilitation", suggesting the establishment of mobile rehabilitation teams. The concept aims at developing a cooperative network linking outpatient with partial hospitalization and inpatient services, and including the physicians at community level. PMID- 9213871 TI - [Assessment methods for vocational integration of disabled persons--from ERTOMIS methods to the IMBA information system]. AB - The background for the development of actual assessments for evaluation of vocational integration of disabled people (i.e., EAM, IMBA) in Germany is described. Resulting perspectives for future procedural approaches are presented. So far, the EAM, Ertomis Assessment Method, approach has gained limited acceptance only. In light of WHO and European impulses, as well as the economic constraints at hand, it remains to be seen whether the IMBA information system will turn out a practical and problem-focussed tool. A critical review is done to provide constructive suggestions for further applications of IMBA. PMID- 9213872 TI - [Tasks of case managers in statutory accident insurance]. AB - The present article gives an outline of vocational rehabilitation in the Federal Republic of Germany from the perspective of the statutory accident insurance scheme. Termed "Berufshilfe", occupational assistance, the services and benefits provided under the scheme also encompass elements of medical and social rehabilitation of persons who have incurred an industrial injury or occupational disease. Given the scheme's comprehensive catalog of benefits and services, a multitude of possibilities result for assistance to eligible workers and their relatives, which are illustrated by examples. Central to the presentation is the work of the "Berufshelfer", the accident insurance agency's case manager for vocational rehabilitation service provision. Along with competence of the accident insurance agency for medical, vocational and social rehabilitation, this case manager is the point of contact for all those involved in rehabilitation in the individual case, hence in particular for the individual affected, his or her relatives, physicians, rehabilitation facilities, employers as well as third parties, moreover being the point of liaison among these various parties and the administration. The primary objective of these entire efforts is to achieve lasting reintegration of people with physical or emotional disablement in employment, career, and society. PMID- 9213873 TI - [Specific and aspecific sports activities of patients with Bechterew's disease]. AB - Regular physical exercise is a major positive influence for the course of ankylosing spondylitis. By means of a standardized questionnaire, a total of 118 patients (78 male, 40 female) with ankylosing spondylitis were asked about their regular physical activity. The patients' mean age was 47 years, and they have been suffering from the symptoms of the disease for (a mean) 17 years. Twenty nine percent of the patients practice disease-specific exercises daily. Thirty nine percent have not received any instructions about the optimal exercises in the previous 5 years. Most information had been given by the physiotherapist at home or at the health resort. Non-specific physical exercises are practiced by half of the patients. Lack of time and lack of motivation are the major factors preventing the patients from greater activity. Deficient mobility was claimed by 37 of the 118 patients to prevent them from practicing sports. Better information, and hence motivation, of the patients are suggested by our findings as the most useful way to foster wider use of physical exercise. PMID- 9213874 TI - [Are there signs of hospitalism in the rehabilitation process?]. AB - The term rehabilitation hospitalism refers to: retardation of rehabilitation progress behind (remaining) abilities. The main causes are: Associate depression (psychoreactive, somatogenic plus endogenic component; usually overlapping), Additional somatogenic factors (heart/circulation, pain, etc), Unfavourable conditions in the surroundings (rehabilitation organisation, family, etc.) Treatment (causal plus symptomatic): Medication, especially psychopharmaceutical, Physiotherapy, Psychotherapy plus organisation plus hygiene plus family therapy No "either/or" but "as well as". PMID- 9213875 TI - [Resolution of the Federal Rehabilitation Council's Governing Board concerning a 2nd bill to refocus self-government and self-responsibility in statutory health insurance]. PMID- 9213876 TI - [Opinion on stage III of the health care structural reform and a 2nd bill to refocus self-government and self-responsibility in statutory health insurance]. PMID- 9213877 TI - [18th World Congress, member assembly 1996 and professional board meeting of Rehabilitation International (RI) 9/13-9/20 in Auckland, New Zealand]. PMID- 9213878 TI - [Economizing potential in rehabilitation--through targetted patient selection]. AB - Major economies need to be made in the health care system. The austerity legislation effective as of January 1, 1997 will result in considerable benefit cuts in the statutory health and pension insurance schemes, with major repercussions also in the field of rehabilitation. Possible devices for realizing potential economies are more precise targetting in patient selection, upgrading social-medical expertise, as well as adjusting rehabilitation more precisely to both patients' needs and economical service delivery. Appropriate tools have been developed by the health insurance scheme towards these ends. PMID- 9213879 TI - [Tramadol (Tramal) in the treatment of rheumatic diseases-- comparative study]. AB - In order to evaluate the efficacy of centrally acting analgesics. In treating rheumatic diseases, tramadol hydrochloride (Tramal Grunental) has been administered to a group of 68 patients (36 women and 32 men), who received 100 mg twice a day during a 10-day treatment. The testing comprised 14 female patients with rheumatoid arthritis, 20 patients (7 women and 13 men) with degenerative (OA) hip and knee diseases and 34 patients (15 women and 19 men) affected by the vertebrogen painful syndrome of lumbar spine. The control group comprised 12 patients (9 women and 3 men) with rheumatoid arthritis using non-steroidal antiinflammatory drugs only, 22 patients (12 women and 10 men) with the OA of the hip and knee, using paracetamol only, and 30 patients (15 women and 15 men) affected by the vertebrogen painful syndrome of lumbar spine, also using paracetamol only. The visual analogue scale has been used in following the pain relief assessments during the therapy. It has thus been observed that the intensity of pain has not been significantly relieved with the acute rheumatic diseases (p > 0.05) in the control group either; that the significant pain relief has occurred with the degenerative (OA) rheumatic diseases (p < 0.05) but not in the control group; while the best analgetic effect of tramadol has been proved on the patients affected by the vertebrogen painful syndrome of lumbar spine (p < 0.01) but was not significant in the control group. During the therapeutic treatment 13 patients (19%), mostly the elderly, experienced side effects, manifested as nausea and the dry mouth. PMID- 9213880 TI - [Dysphagia due to cervical spine involvement in ankylosing spondylitis]. AB - Here we present a case of a patient with a massive ossification of annulus fibrosus and longitudinal ligament at the level C3-C4, C4-C5 and C5-C6, also called the Van Swaay bridge. One can observe a propulsion of the pharingeal tissue at the level of the massive Van Swaay bridge between C5 and C6 which caused pressure and dysphagia. PMID- 9213881 TI - [Hypertrophic osteoarthropathy in hemiparesis]. AB - A case of concomitant hypertrophic osteoarthropathy and hemiparesis is presented. The patient had several attacks of cerebrovascular insult with hemiparesis of both the left and right sides. After the last attack he developed clubbed fingers and later periostal reaction on the long bones. The case is interesting and rare, particularly as we have not found similar data in literature. PMID- 9213882 TI - [Incorrect diagnosis of peripheral arthritis in ankylosing spondylitis]. AB - We present a case of female patient who's been treated 28 years, under the diagnosis of rheumatoid arthritis because of the symmetric polyarthritis. After 28 years radiography of sacro-iliac joints and thoracolumbar vertebra was taken and showed changes typical for ankylosing spondylitis with asymmetrical affection of peripheral joints and with irreversible changes only in knees joints. Rheumatoid factor in serum was always negative, and the patient has also an iridocyclitis. We conclude that the diagnosis is surely ankylosing spondilitis. PMID- 9213883 TI - [Antigenic and genomic characterization of Herpes simplex virus isolated from double genital infections]. AB - Two Herpes simplex viral serotypes, HSV type 1 and HSV type 2 may cause genital herpes. The aim of this study was to perform a genetical analysis and characterization of virus isolated from four patients with double genital herpetic infections. In eleven viral isolates, the cytopathic effect in Vero cells was studied, the antigenic type was determined using monoclonal antibodies and genomic analysis was performed with Eco RI, Hind III and Bgl II enzymes. Five viral isolates generated a diffuse and 6 a localized cytopathic effect. Monoclonal antibodies identified four HSV-1 and seven HSV-2. Genomic analysis had concordant results. Four HSV-1 were obtained, with different genomic patterns within them, three were different to the standard North American strain. The seven HSV-2 obtained had three different types of electrophoretic profiles, that were different to the standard North American strain. It is concluded that the genomic and antigenic analysis allowed the detection of herpetic genital infections caused by herpes virus type 1 and 2 in the same individual and the identification of herpes virus strains with distinct regional characteristics. PMID- 9213884 TI - [Patients with rheumatoid arthritis: study of the correlation between density of glucocorticoid receptors in synovial cells and clinical response to steroidal treatment]. AB - BACKGROUND: The target cellular response to glucocorticoids is proportional to the concentrations or affinity of specific receptors to these substances. AIM: To look for a correlation between glucocorticoid receptor concentrations in synovial wall cells and the clinical response to steroidal treatment in patients with rheumatoid arthritis. PATIENTS AND METHODS: Twenty eight patients with rheumatoid arthritis were studied. Each subject was subjected to a synovial biopsy, in which a dry radioautographic technique for diffusible compounds was used. Patients were treated afterwards with three 500 mg intravenous pulses of methilprednisolone. RESULTS: A mean of 44.8% of synovial cells (range 30.1-62.8%) had binding sites for 3H dexamethasone. All patients had a significant clinical improvement after methylprednisolone. Multiple regression analysis did not show a correlation between clinical response and glucocorticoid receptor concentration. CONCLUSIONS: The lack of association between glucocorticoid receptor concentrations and clinical response could be due to the large steroid dose used, that saturated all available receptors. PMID- 9213885 TI - [Effects of casein on small intestine motility in patients with liver cirrhosis]. AB - BACKGROUND: Fasting small bowel motor abnormalities have been described in cirrhotic patients. AIM: To investigate the effects of orally administered casein in small bowel motor activity in cirrhotic patients. PATIENTS AND METHODS: Six healthy subjects and 23 cirrhotic patients were studied. Motility of the upper part of the small bowel was studied by means of perfused manometric catheters, during a basal period and after infusion of 30 g of casein in the stomach. RESULTS: Basal recordings showed a cyclical activity in all healthy subjects and in 9 cirrhotics (further considered as group 1). In 14 patients, basal cyclical activity was absent and were further considered as group II. Frequency of contractions after casein infusion was higher in group II patients than in group I and healthy controls (2.2 +/- 0.3, 1.3 +/- 0.2 and 0.9 +/- 0.2 cpm respectively). Intestinal motor index after casein infusion was also higher in group II. There was a progressive decline with time in motor activity after casein infusion, being maximal at 45 min, in healthy subjects and group I. This decline was not observed in group II. CONCLUSIONS: Patients with cirrhosis have an altered small bowel motor response to casein. Since group II of patients had a greater percentage of Child C patients, the abnormalities seem to be related to the degree of liver failure. PMID- 9213886 TI - [Approximation to the dynamics of meningococcal meningitis through dynamic systems and time series]. AB - Meningococcal meningitis is subjected to epidemiological surveillance due to its severity and the occasional presentation of epidemic outbreaks. This work analyses previous disease models, generate new ones and analyses monthly cases using ARIMA time series models. The results show that disease dynamics for closed populations is epidemic and the epidemic size is related to the proportion of carriers and the transmissiveness of the agent. In open populations, disease dynamics depends on the admission rate of susceptible and the relative admission of infected individuals. Our model considers a logistic populational growth and carrier admission proportional to populational size, generating an endemic dynamics. Considering a non-instantaneous system response, a greater realism is obtained establishing that the endemic situation may present a dynamics highly sensitive to initial conditions, depending on the transmissiveness and proportion of susceptible individuals in the population. Time series model showed an adequate predictive capacity in terms no longer than 10 months. The lack of long term predictability was attributed to local changes in the proportion of carriers or on transmissiveness that lead to chaotic dynamics over a seasonal pattern. Predictions for 1995 and 1996 were obtained. PMID- 9213887 TI - [Hip fracture: a case-control study in the metropolitan region I]. AB - BACKGROUND: Hip fracture is a preventable cause of disability among elderly people AIM: To study factors associated to hip fractures in Chile. PATIENTS AND METHODS: Patients admitted to seven public hospitals in Chile, with hip fracture not due to major accidents, were considered as cases. To each, at least one age and sex matched hospitalized control, without or neoplastic diseases, was assigned. All patients were subjected to an inquiry, using an instrument devised by the WHO. RESULTS: Two hundred sixty eight cases and 501 controls were studied. Cases and controls has similar educational and labor histories. The right hip was fractures in 47% of cases and the left in the rest. Compared with controls, cases had a higher body mass index, loss of height during life, rate of hysterectomy, incidence of smoking and a lower consumption of diary products. Cases had higher risk of falls inside their homes and controls outside. CONCLUSIONS: The obtained information may contribute to the development of preventive measures and reduce the public health impact of hip fracture. PMID- 9213888 TI - [Study of volume and kinetics of gallbladder emptying in non-pregnant, pregnant, and puerperal women]. AB - Gallbladder stasis apparently plays an important role in the patho-genesis of cholelithiasis during pregnancy. On the other hand, gallstones diagnosed immediately after delivery disappear spontaneously during late puerperium in one third of these patients. Gallbladder emptying was assessed by biliary scintigraphy and ultrasonography in normal, nulliparous volunteers. The two methods had an excellent correlation: thence, we used ultrasonography to determine gallbladder volume and contraction in pregnant and puerperal women. Fasting and postprandial residual volumes were significantly larger during pregnancy, while the kinetics f gallbladder emptying was similar in nulliparous and pregnant women. During puerperium, gallbladder volume returned to the values observed in nulliparae; but the kinetics of emptying was significantly faster, suggesting an increased sensitivity of gallbladder muscle to physiologic stimuli. PMID- 9213889 TI - [Anatomopathological study of 86 gastric adenomas. Experience in 14 years]. AB - AIM: To analyze the clinical presentation, pathological aspect and treatment of gastric adenomas. PATIENTS AND METHODS: Retrospective analysis of 75 patients aged 26 to 88 years in whom a gastric adenoma was diagnosed. RESULT: Seventy one patients had elevated endoscopical lesions and two had depressed or flat lesions. Ninety percent of lesions were located in the gastric antrum. Pathological study detected 6 focal carcinomas within the adenomas and 5 concomitant carcinomas located elsewhere in the stomach. Fifty four patients were subjected to endoscopical resection. Among patients with focal carcinomas, a gastrectomy was performed in four and endoscopical resection in two. CONCLUSIONS: Gastric adenomas must be considered in the differential diagnosis of gastric elevated lesions and may be confused in early gastric cancer. There is a histological resemblance between adenomas and gastric dysplasia described by several authors though only in our cases and in the Japanese literature the adenoma is referred to as mostly a polypoid sessile lesion. PMID- 9213890 TI - [Late centronuclear myopathy: autosomal dominant form]. AB - We report a family with three generations affected by an autosomal dominant centronuclear palsy. This gene is characterized by ptosis that begins in childhood and a slowly progressive weakness that starts in the second decade of life, involving face, neck and limbs. In this stage, muscle pan associated to exercise or cold muscle spasms may appear. The gene is expressed with differing intensity in each individual. Myopathic electromyographic alterations are only found in functionally impaired subjects. Muscle biopsy shows type I fiber atrophy and central nuclei in a high percentage of fibers, specially in type I fibers. PMID- 9213891 TI - [Seronegative arthritis: reexamination of the initial diagnosis and prognosis after 10 years]. AB - Seronegative arthritides are a heterogeneous group of diseases that includes rheumatoid arthritis with negative rheumatoid factor. Between 1980 and 1984 we studied 38 patients with seronegative arthritis. Thirty of these patients were reassessed in 1994 after 9 to 20 years of evolution. Seventeen patients had a diagnosis of seronegative rheumatoid arthritis; this diagnosis was maintained in 12, changed to seropositive rheumatoid arthritis in three, to psoriatic arthritis in one and connective tissue disease in one. Thirteen patients had a diagnosis of undifferentiated arthritis; in 1994 the diagnosis was maintained in three, seven patients were diagnosed as having a spondyloarthropathy, two as having a reactive arthritis and one as having a connective tissue disease. In 1994, nine patients fulfilled the 1991 criteria for spondyloarthritis and six of these did so on admission. Six of 12 patients with seronegative rheumatoid arthritis had an active disease or used antiinflammatory drugs and 64% had erosions on hand X ray examinations. These figures are in contrast with the enign evolution classically attributed to this disease and agree with recent reports. The usefulness of classification criteria for rheumatoid arthritis and spondyloarthritis in the initial assessment of patients with seronegative arthritis is emphasized. PMID- 9213893 TI - [Simple juxtathyroid cyst. Analysis of the possible origin of the lesion]. AB - We report a 34 years old female who presented with a simple juxtathyroid cyst. Its content was translucent with a mPTH concentration 30 higher than in blood. This finding lead to the suspicion of a parathyroid cyst; however there were no laboratory evidence of hyperparathyroidism. Two years later and after repeated needle drainages, a surgical cystectomy was made. The immunohistochemical study of the samples was intensely positive for synaptophysin, a parathyroid tissue marker. Surprisingly, thyroglobulin was also found in some cyst wall cells. PMID- 9213894 TI - [Gene therapy: facts and fiction]. AB - The great advances in molecular biology have encouraged the treatment of diseases through gene intervention. According to present knowledge, all diseases somehow have their origin or are related to genes. Different stages must be overcome to intervene in genes: a) Identification of the gene to intervene. Methodologies have been developed and a number of genes identified. b) Codification of its structure. The technology for this purpose is already known and the process has been automated. c) Locate the adequate vector that, without damaging the cell, is capable to introduce in nucleus the genetic material. There are many studied alternatives. However the ideal vector, that would select the target cells, be risk-free and guarantee prolonged results, has not been found yet. d) To achieve the gene expression, codifying the respective protein or its inhibition, according to the case. Some success has been obtained in this field, but technology has to be improved to obtain a uniform and satisfactory response. Advances achieved in the last years have been impressive and technologies will presumably continue to improve. PMID- 9213892 TI - [Liver neoplasms in patients with glycogen storage disease. Diagnostic and therapeutic problems]. AB - In a family composed by eight brothers, five had a type I glycogen storage disease and three presented with liver tumors complicated with hemorrhage or malignant transformation, during the follow up. The periodic ultrasonographic control allowed the early diagnosis of these neoplasms. We describe the clinical picture and treatment of two patients. Metabolic alterations were corrected during the preoperative period with parenteral nutrition. Resection was successful in both cases. It is concluded that a close follow up and early treatment of these lesions is effective and avoids complications. In patients with multiple lesions and severe metabolic alterations, liver transplantation is the treatment of choice. PMID- 9213895 TI - [Cytokines, growth factors, and metabolic bone disease]. AB - Cytokines are polypeptides that bind to membrane receptors and may act in an endocrine, paracrine or autocrine way. Several cytokines and growth factors may be produced by bone cells, stored in the matrix or act on them. Osteoclasts derive from the bone marrow stem cell and, as monocytes, belong to the family of tissue macrophages. Their specific function is bone resorption. Interleukin 1, 6 and 11, transforming growth factor and tumor necrosis factor stimulate osteoclast mediated bone resorption. Interleukin 1 is the most potent bone resorption agent and seems to be identical to osteoclast activation factor, identified in multiple myeloma. The role of interleukin 1, 6, 11 and tumor necrosis factors in postmenopausal osteoporosis triggered by the fall in estrogen levels, has not been well defined yet. Cytokines that increase bone formation are insulin like growth factors I and II, transforming growth factor, platelet derived growth factor and bone morphogenic proteins. Probably, tumor necrosis factor and interferon-gamma have a depressor effect on bone formation. Cytokines and growth factors, liberated from bone cells or from the matrix during osteoclastic work, could be the signals responsible for coupling bone formation and resorption. PMID- 9213897 TI - [Obstructive jaundice, infrequent manifestation of autosomal dominant polycystic disease]. PMID- 9213896 TI - [The imperative of medical involvement in the implementation of quality control systems]. AB - Changes in health care delivery in Chile over the last years include rising costs and the relative increase of prepaid private insurance programs. It is expected that there will be increasing pressures on health providers to decrease cost while maintaining or improving quality. A series of common clinical problems are reviewed (hernia repair, appendicitis, gallstone disease, trauma, preoperative evaluation) to demonstrate that using a scientific method-continuous quality improvement- and considering the local socioeconomic reality, this challenge can be met. Emphasis is made on the need for physicians to participate in such a process and on the teaching of these concepts. Medical societies and schools should consider these health care developments and adopt to this changes. PMID- 9213898 TI - [Image of the month. Premalignant lingual leukoplakia]. PMID- 9213899 TI - [Pharma-clinics. How I treat... A patient with orthostatic hypotension]. PMID- 9213900 TI - [Clinical case of the month. Osteoporosis and beta-thalassemia]. PMID- 9213901 TI - [Obstetric perspectives: consensus of the gynecology department of the University of Liege. Document of the 3rd cycle studies, October 96]. PMID- 9213902 TI - [Vaccination in children]. PMID- 9213903 TI - [Ophthalmological follow-up of patients receiving corticotherapy]. PMID- 9213904 TI - [Surgical treatment of tricuspid valve insufficiency. Carpentier's annuloplasty]. PMID- 9213905 TI - [Transsexualism: general considerations and management]. PMID- 9213906 TI - [The family of natriuretic peptides]. PMID- 9213907 TI - [Dolly or the best in the world--a prelude to human cloning?]. PMID- 9213908 TI - [How I explore ... At the coronary level, a patient prior to noncardiac surgery]. PMID- 9213909 TI - [What is your diagnosis? Glomus tumor]. PMID- 9213910 TI - [Evidence based medicine]. PMID- 9213911 TI - [Evidence-based medicine: approach to a more rational medicine. Arbeitsgemeinschaft Cochrane Collaboration Schweiz]. PMID- 9213913 TI - [Concepts important in the interpretation of therapeutic studies. Absolute risk reduction (ARR), relative risk reduction (RRR), number needed to treat (NNT)]. PMID- 9213912 TI - [Does alendronate reduce the risk of fractures in postmenopausal women with osteoporosis?]. PMID- 9213914 TI - [Cholesterol crystal embolisms: an exciting search for "pearls"]. AB - This article is meant to increase the interest in an often forgotten clinical entity. Cholesterol emboli are in the majority of cases only diagnosed at post mortem examination. Even though the triad livedo reticularis, renal failure and eosinophilia constitutes its most prominent feature, the variable clinical manifestations of this disorder with multiorgan involvement ("pseudovasculitis") make the search for cholesterol crystals particularly exciting. The discovery of 10 cases of cholesterol emboli over 2 years in a regional hospital's internal medicine department demonstrates that this occurrence is not rarely and that its accurate identification can be particularly relevant. It is important to recognize this disease since it is often iatrogenic, affects elderly people with atherosclerosis of the large vessels and causes high morbidity and mortality. PMID- 9213915 TI - [Sudden death under anticoagulation for pulmonary embolism]. PMID- 9213916 TI - [Liver injury under tuberculostatic treatment]. AB - We report the case of a patient with nausea, loss of appetite and increase of the aminotransferase levels to eight times the upper normal limit occurring two weeks after she was started on isoniazide, rifampicine and pyrazinamide for treatment of tuberculosis. Isoniazide is the most likely cause of liver injury occurring during combined antituberculosis therapy, whereas pyrazinamide or rifampicine are only rarely responsible. The case presented is used to review and compare the different recommendations concerning the monitoring of patients receiving antituberculous therapy and the clinical management of patients developing liver injury. PMID- 9213917 TI - [Cytomegalovirus colitis in AIDS]. PMID- 9213919 TI - Bibliography of recent literature in sleep research. PMID- 9213918 TI - [Benign monoclonal lambda type IgG-gammopathy]. PMID- 9213920 TI - Letter from the editor: first, a little mamba . PMID- 9213921 TI - [Decision-making by the surgeon during operations]. PMID- 9213922 TI - [Injury of the extrahepatic bile ducts in multiple injuries]. AB - During the ten-year period between 1982-1991 the authors operated in the Institute 12 casualties after perforation and avulsion of the gallbladder, 8 patients on account of injuries of the hepatocholedochus and 15 patients where after injury posttraumatic acute cholecystitis developed. The authors evaluate critically the asset of new diagnostic preoperative procedures for assessment of early indication of surgery. In the surgical tactics they prefer reconstruction procedures which prevent such serious postoperative complications as relapsing cholangitis or stricture with cholestatic affection of the liver [1,3]. PMID- 9213923 TI - [Obturator bypass--a method of extra-anatomic reconstruction in the region of the femoral trigone]. AB - The authors present the case-history of a patient admitted on account of haemorrhage in the right groin during an infectious complication after an aortobifemoral bypass. The authors selected an obturator bypass as the method of choice for extraanatomical reconstruction and as an example how to resolve this serious complication in vascular surgery. PMID- 9213924 TI - [Benign lesions of the liver and their surgical treatment]. AB - The authors submit a group of 17 patients with benign hepatic lesions operated during the past five years. Most frequently haemangiomas were encountered, less frequently adenomas and focal nodular hyperplasia. It was revealed that accurate diagnosis is possible even when sonography, CT, MRI and even scintigraph (SPECT) are used only in haemangioms. When indicating surgery the clinical symptomatology, uncertain diagnosis and difficulty and risk of surgery itself must be taken into account. PMID- 9213925 TI - [The role of the portosystemic shunt in modern treatment of portal hypertension]. AB - The authors discuss the importance of portosystemic anastomoses in contemporary treatment of portal hypertension, in particular in relation to TIPS. As a basis they use their own experience with 28 portosystemic anastomoses during the last five years and conclude that a correctly indicated anastomosis has certain advantages over TIPS. The basic requirements for indication of surgical peripheral portosystemic anastomoses are: 1. stage Child A (or onset of B), 2. posthaemorrhagic conditions, 3. satisfactory general condition, 4. stabilized liver disease and planned surgery. PMID- 9213926 TI - [Use of collagen and gentamycin in the treatment of chronic soft tissue lesions]. AB - The authors describe two case-histories where Garamycin-schwamm was used in the treatment of a chronic fistula and cavities in the subphrenium resulting from treatment of a subphrenic abscess, and in the pelvis minor as a result of a fistula which developed after establishment of an ileoanal intestinal pouch. They confirm the experience of authors abroad with the treatment of chronic cavities in soft tissues as a result of operations, which so far had to be treated by other methods. PMID- 9213927 TI - [Transverse laparotomy closed with continuous absorbable loop sutures]. AB - The authors emphasize the advantages of a transverse incision which are beyond doubt, because they provide the surgeon not only with an excellent bilateral view of the operated area, but what is most important, they have the significantly least negative effect on respiratory functions and the composition of blood gases, which is of major importance in patients with chronic respiratory failure. Due to the much smaller retraction forces the incidence of postoperative dehiscences and hernias is smaller. It is an incision which interferes least with the innervation of the abdominal wall and thus is not only less painful but has also better healing parameters. Its closure by a continuous, absorbable, loop-on mucosa double suture is not only simple, but what is most important, it is reliable and associated with a minimum of postoperative complications. PMID- 9213928 TI - [Liver metastasis in colorectal carcinoma]. AB - The authors describe the method and therapeutic results of liver metastases of colorectal carcinoma in a group of 14 patients followed up for 2 to 33 months. In these patients in five instances (35.7%) resection with subsequent intraarterial chemotherapy (HAI) was possible. In nine instances (64.3%) a chemoport was introduced, as resection was impossible due to the size of the lesion. For checking of the site and patency of the arterial chemoport, the authors recommend the use of port scintigraphy, a method which is accurate, accessible and simple. PMID- 9213929 TI - [Enteral nutrition--nasojejunal tube or percutaneous jejunostomy?]. AB - As many as 60% patients develop malnutrition during the postoperative period. Enteral nutrition can prevent its development or at least mitigate its manifestations. The author mentions the advantages, disadvantages, indications and contraindications of enteral nutrition. He mentions tho advantages and disadvantages of two routes of establishment of enteral nutrition, i.e. by a nasoenteral tube and by puncture stomy. Of many possible procedures he describes experience with a nasojejunal tube and with puncture jejunostomy. He deals with complications he recorded with both methods and presents two case-histories of less common application of puncture jejunostomy. The author prefers the use of puncture jejunostomy to that of a nasojejunal tube and recommends its use after surgery of the upper GIT and in acute necrotizing pancreatitis. PMID- 9213930 TI - [Results of resection of a stage IIIA/N2 non-small cell bronchogenic carcinoma]. AB - The authors analysed a group of 80 patients who were operated in 1985-1990 on account of non-small cell carcinoma of the lungs in stage IIIA with affection of the ipsilateral mediastinal nodes. The patients were not treated by neoadjuvant chemotherapy and systematic dissection of the mediastinal nodes was not performed. The results of five-year survival in the group of patients with affection of the mediastinal nodes (N2) were compared with those in the group of patients without affection of the mediastinal and hilar nodes (NO). Patients with affected N2 nodes who survived five years were significantly fewer than patients with negative mediastinal and hilar nodes. The probability of five-year survival in N2 was 15.8%, in patients with NO 28.0%. From data in the literature it is known that neoadjuvant chemotherapy and subsequent complete resection of the lung with the tumour combined with dissection of the mediastinal nodes may improve long-term survival after surgery. The authors assume that introduction of the described methods in their department will improve postoperative results. PMID- 9213931 TI - [Laparoscopic appendectomy using the out-transumbilical method-- personal experience]. AB - The author present an account of his experience with 80 laparoscopic appendectomies using the method out (LAPPE-out). It is a method combining the advantages of laparoscopy, i.e. mininvasiveness and the possibility to view the abdominal cavity with the certainty of treating the base of the appendix under direct visual control. As to the length of the operation and its pretentiousness it matches classical appendectomy (class. APPE), as regards postoperative comfort and possibilities of laparoscopy it is clearly superior to the latter. PMID- 9213932 TI - [Radical surgery of stage II primary nonresectable colonic tumors]. AB - The authors present an account on six patients with a primarily non-resectable tumour of the colon where radical resections were made in the second stage. Based on their experience they recommend an active approach to patients with primarily non-resectable tumours of the colon who have no signs of dissemination of the tumour. In these patients they recommend not only dispensarisation but also adjuvant chemotherapy and radiotherapy. In case of regressing tumourous infiltration they recommend resection of the tumour in the second or if necessary third stage. PMID- 9213933 TI - [Laparoscopic reconstruction in inguinal hernias]. AB - The author describes his experience with laparoscopic plastic operations of inguinal hernias by means of a prolene net. This tension-free method makes rapid convalescence and return to normal life possible. A very important advantage as compared with classical procedures is the substantially lower percentage of relapses. Advanced experience (639 plastic operations) by the TransAbdominalPrePeritoneal method justify the following conclusions: The operation is much less painful, it is cheaper (if we take into account the costs of a longer stay in hospital and longer work incapacity after classical plastic operations of inguinal hernias) and there is a lower percentages of relapses (0.6%). The author has the most favourable experience using a net with a preperitoneal transabdominal approach. The very good results obtained with 639 operations are a reason for optimism. The development of new techniques will, no doubt contribute to a clearer view of this complicated problem. PMID- 9213934 TI - [Transjugular intrahepatic portosystemic shunt in the treatment of complications in portal hypertension]. AB - Transjugular intrahepatic portosystemic shunt (TIPS) is a side-to-side portocaval shunt for threatening complications of portal hypertension. The purpose of this study was to evaluate in first 33 patients indicated for TIPS insertion in our institution the efficacy, complications, and mortality. Indication was failure of sclerotherapy or ligation in control either of acute (n = 4) or repetitive (n = 25) variceal bleeding and refractory ascites (n = 4). The technical success rate was with 70% (21/30) lower than expected, but the complication rate was also very low. There were no fatal complications, only one subcapsular liver hematome, and in one patient repetitive punction of biliary tract. The 30-days mortality was 10% (2/21) and rebleeding was 15% (3/20), caused always by thrombosis of the shunt. TIPS seems to be a promising therapeutic procedure after failed endoscopic therapy of esophageal varices without the mortality and morbidity of an open surgical procedure. Recent indications for TIPS are acute variceal hemorrhage refractory to endoscopic treatment and recurrent variceal bleeding despite sclerotherapy or band ligation. Promising seems to be TIPS insertion in the treatment of refractory ascites. PMID- 9213935 TI - FIGO Committee for the Study of Ethical Aspects of Human Reproduction: Ethical aspects in the management of the severely deformed fetus. PMID- 9213936 TI - [A new dimension--proprioceptive training of the upper extremities and trunk. Case report of a 13-year-old tennis player after shoulder dislocation]. PMID- 9213937 TI - [Treatment of osteoporosis by activation of endogenous physiological regenerative capacities]. PMID- 9213938 TI - [Proprioception of the shoulder joint in young tennis players]. AB - In 60 adolescent volunteers aged 8 to 16 years we assessed the proprioceptive capability of the shoulder complex by an angle reproduction test. The purpose of the study was to evaluate the proprioceptive capability of adolescent tennis players. 40 were tennis players, 20 non tennis players served as a control group. Documentation of the reproduced angle was performed by a motion analyzing system with passive markers. Angle reproduction of all volunteers was best in the midrange of motion (100 degrees flexion, 100 degrees abduction, neutral rotation in 90 degrees abduction). The worst results were documented below shoulder level (50 degrees flexion, 50 degrees abduction, internal rotation in 90 degrees abduction). A correlation to sex or dominant extremity could not be found. Subjects older than 12 years showed a tendency for better angle reproduction compared to the younger subjects. Tennis players older than 12 years demonstrated significant better capabilities for angle reproduction in some movements of the shoulder complex. PMID- 9213939 TI - [Results of surgical treatment of radio-humeral epicondylopathy]. AB - We examined extent and affection of an assumed neuromuscular transmission disorder by performing a prospective clinical study on 75 patients with therapy resistant radiohumeral epicondylopathy. Before operation, we electromyographically diagnosed an increased rate of polyphasic potentials of the long wrist extensors as well as a prolonged motor latency of the respective muscles. Corresponding to a hereby implied damage to the distal part of the motor neuron, disordered neuromuscular recruitment combined with a reduced maximum strength and -elasticity could be proven. Both effects were significantly reversible (p < 0.001) through operative intervention. We found a significant correlation (corr < 0.90) between the normalization of the motor latency and increased strength. Subgroups were formed depending on different pre-operative diagnostic efforts and differing redicality regarding the performed soft-tissue operation, thus the clinical validity of the findings diagnosed in the anatomic and neurophysiologic part of the study was additionally examined. It was proven that the failure rate varies between 10% and 30%, depending on the radicality of tenotomy, which could be interpreted as a general indication for complete extensor carpi radialis brevis tendon release. In this connection it is remarkable that the clinical result of an electromyographically localized damage in the area between epicondyle and arcade of Frohse could not be improved through open neurolysis. Dealing with strictures located on the proximal side of the epicondyle on the other hand, this technique seems to play an important role for recurrence prophylaxis. PMID- 9213940 TI - [Sports-induced acute epicondylitis of the elbow and conservative therapy]. AB - 48 patients with an acute lateral and/or medial epicondylitis of the elbow, induced by sporting activities, were treated in a prospective, controlled, double blind, and randomised clinical study over a period of one to two weeks in a standard manner, either with taping and the oral NSAID proglumetacine b.i.d. (26 patients: 8 women, 18 men; mean age of 37.8 years) or with taping and an oral placebo b.i.d. (22 patients: 11 women, 11 men; mean age of 40.0 years). The course of complaints was followed on the basis of a functional index (created according to Lequesne). Taking into account the prior definition of efficacy, at the end of the 1st week of all 22 NSAID group patients, enrolled and treated according to the protocol, there were already 21 responders and of all 19 corresponding placebo group patients, there were 12 responders. The difference is statistically significant (p = 0.0157). The rate of efficacy was 95.5% in the verum (plus taping) group and 63.2% in the placebo (plus taping) group with a further amelioration in both groups at the end of the study. Safety was comparably good in both the verum as well as the placebo group. PMID- 9213941 TI - [Surgical treatment of chronic ankle joint instability--many variations in German clinics. Analysis of a 1994 German survey]. AB - A questionnaire was mailed to 400 orthopaedic department chairpersons in Germany listed by the "Deutsche Gesellschaft fur Unfallchirurgie" to evaluate the current surgical approach to chronic ankle instability. The aim of the survey was to analyze and to classify the reconstruction method of the individual institution. Questionnaires were returned from 267 hospitals (66.7%). The number of ligament reconstructions performed at individual institutions ranged from 1 to over 100 (mean 17.5). The most frequently used procedures of primary choice for reconstruction of the ligaments are: periosteal flaps (45.3%), anatomical repair (23.6%), different tenodeses (20.6%), and free anatomical implants of autologous or exogenous material (10.5%). 50% of the participants use reconstructions in which the injured ligament is repaired secondarily without augmentation or with periosteal flaps. The remaining part of the German surgeons prefers tenodeses or various other procedures. It is interesting to note that there is a high number of different procedures not namely known in the literature which are considered as individual modifications. With better knowledge of the function of the single ligament and general exposure of the ligaments for simple overlap and direct suture, the rate of primary non-anatomical reconstructions may be decreased in the future. PMID- 9213942 TI - [Effect of external stabilizing agents of the ankle joint on sports motor capacities in one legged jumping]. AB - In order to document sport specific skills, 23 athletes with functionally unstable ankle joints as well as 18 healthy volunteers were evaluated by performing a single leg jumping test. For external stabilization of the ankle an Aircast-Brace, a Ligafix-Air-Brace, a Malleoloc-Brace as well as a tape bandage were applied. There was no negative influence on the jumping performance of the tested stabilizing devices in the uninjured ankle joints. There was also no significant difference between the devices. For functional unstable ankle joints most of the devices showed significant improvement of jumping performance. The best results were achieved by the Aircast-Brace, followed by the Malleoloc-Brace, the Ligafix-Brace and the tape bandage. Again, there was no significant difference between these devices. While the reaction time showed no difference in all test situations, the time for stabilizing the ankle joint was significantly worse in those ankle joints without a brace. CLINICAL RELEVANCE: For athletic activities, which are dominated by movement patterns comparable to the applied jumping test, the tested stabilizing devices seem to have no negative effect on sports specific capabilities. PMID- 9213943 TI - [Isolated pelvic vein thrombosis in a young marathon runner]. AB - The case of a 20-year old female patient who developed an isolated thrombosis of the pelvic vein after a marathon run, is presented. After surgical treatment and anticoagulation for one year, complete restitution was seen. The possible relation between marathon running and venous thrombosis without other risk factors is discussed. PMID- 9213944 TI - [Sports medicine aspects of gymnastics]. AB - 926 injuries were collected and analyzed through a retrospective questionnaire survey among 289 competitive gymnasts. On the average a male/female athlete suffered 0.35/0.33 injuries per year. The greatest number of injuries occurred during floor exercise. Male gymnasts had a high average number (52%) of upper body injuries. The wrist was most often affected (15.4%). The females suffered more injuries in the lower body (52%). The ankle was the most frequently injured anatomical site (28.5%). A comparison between different level gymnasts shows that a more intensive training does not lead to different injuries. When comparing injuries to expose time fewer occurred in the group of high level gymnasts. PMID- 9213945 TI - [The early diagnosis of hypertension]. PMID- 9213946 TI - [The problem of the accurate noninvasive diagnosis of left ventricular myocardial hypertrophy in essential hypertension: the role of magnetic resonance tomography]. AB - MR-tomography was compared to echocardiography for efficacy in diagnosis of left ventricular hypertrophy in 55 patients with stage II essential (WHO criteria). Left ventricular myocardial mass estimated at echocardiography appeared significantly greater than this mass estimated at MR tomography: 244.0 +/- 9.0 g (MRT) and 359.8 +/- 15.6 g (echo-CG), respectively (p < 0.001). PMID- 9213948 TI - [A decrease in left ventricular hypertrophy and the dynamics of the parameters of circadian BP monitoring under the influence of ramipril in patients with essential arterial hypertension]. AB - To evaluate factors of action of ramipril, an inhibitor of angiotensin-converting enzyme, which may induce a decrease in left ventricular hypertrophy (LVH), 45 LVH patients aged 21-53 years with mild and moderate essential hypertension have underwent echo-CG determination of left ventricular mass and 24-h monitoring of arterial pressure (AP). Multiple regression was used to examine prognostic significance of such parameters as age, sex, height, weight, duration of the disease, mean 24-h AP, its variability and the degree of nocturnal fall. It was found that ramipril reduced LVH irrespective of the hypotensive effect. PMID- 9213947 TI - [Doppler echocardiography in assessing the effect of the beta 1-selective adrenoblocker acebutolol on left-ventricular diastolic filling in hypertension patients]. PMID- 9213950 TI - [The deep veins of the lower extremities studied by ultrasonic dopplerography in patients with anomalously positioned chordae tendineae]. PMID- 9213949 TI - [The possibilities of using ambulatory stress echocardiography with transesophageal atrial stimulation]. AB - Stress echocardiography (duplex regimen echocardiography combined with transesophageal pacing) was performed outpatiently in 64 subjects. Among them there were patients with verified angina of effort and ischemic heart disease suspects. The test reached diagnostic criteria in 85% of the examinees. Significant signs of coronary insufficiency were recorded in 12% of patients with doubtful evidence obtained at bicycle ergometry. This method is proposed as an alternative (in addition to stress ECG tests) variant of outpatient diagnosis of coronary insufficiency in doubtful cases and to assess coronary reserve in patients with verified ischemic heart disease. PMID- 9213951 TI - [The importance of the Holter ECG monitoring of patients in the acute period of an ischemic stroke]. PMID- 9213952 TI - [The possibility for the presumed detection of the obstructive lesion of the coronary arteries in stenocardia patients by the data from circadian ECG monitoring and stress tests]. AB - Whether 24-h ECG monitoring (ECGM) may be used in detection of multiple arterial obstructions in anginal patients and to compare diagnostic efficacy of ECGM and bicycle exercise (BE) in prediction of 2 or 3 lesions was investigated in 137 patients subjected to ECGM and selective coronarography. 117 of them did bicycle exercise. The number of episodes of painful and painless myocardial ischemia in ECGM was related with the extension and depth of the coronary flow obstruction. The overall index of myocardial ischemia has been estimated warranting higher probability of detection of subjects with obstruction in 1 or more coronary arteries. Sensitivity and prediction value of ECGM was similar to those of BE, but the latter was more specific. PMID- 9213953 TI - [The importance of ambulatory ECG monitoring for determining the prognosis of patients with stable stenocardia]. AB - 149 patients with coronary artery disease and stable angina pectoris underwent coronary angiography and had coronary artery stenosis over 50 per cent. All the patients were also subjected to 24-h Holter monitoring at primary examination and 12-18 months after it. Typical ischemic ST changes were defined by transient horizontal or descending ST depressions > 1.0 mV (measured 80 ms after the J point) lasting at least for a minute. 75 (50.3 per cent) patients had episodes of silent myocardial ischemia. The course of the disease was assessed in follow-up period of 12-18 months. Four variants of the course were determined: cardiac events (16 patients), the disease progression (33 patients), a stable course (75 patients), clinical remission (25 patients). A significant correlation between the occurrence, the slope and duration of silent ischemia, the data of selective coronary angiography and clinical course of ischemic heart disease was established. Cardiac events occurred in 87.5% of the patients with silent myocardial ischemia who had total ischemic burden 30 minutes or more and/or ST segments decrease 3.0 mm and more during heart rate less than 100 beat-min. The stable course was registered in patients with silent ischemia or without it with similar frequency. Clinical remission of angina pectoris in the patients with silent ischemia was observed rarely. The results of this study demonstrate that silent ischemia is an important prognostic factor. PMID- 9213954 TI - [The constitutional and genetic characteristics of patients with the sick sinus syndrome]. PMID- 9213955 TI - [The diagnosis and elimination of disorders in cardiac electrostimulation in patients with an artificial pacemaker]. AB - In 378 patients with artificial pacemaker (AP) 10-year follow-up revealed disturbances of continuous pacing and cardiac arrhythmia. Diagnostically significant were: ECG, Holter ECG monitoring, functional AP control. Cardiac arrhythmias were spontaneous and pacing-induced. Programmed AP corrects complications of cardiac pacing and arrhythmia without surgical intervention. Regular technical control of the stimulation system providing early diagnosis of AP defects is an important component of check-ups of patients with AP. PMID- 9213956 TI - [The runaway pacemaker syndrome during electrical ventricular stimulation of the heart]. AB - The paper reports a case of escaping pacemaker syndrome in a female with continuous endocardial ventricular pacing. The syndrome was provoked by defects in the electronic scheme because of weakening of pacemaker's battery. PMID- 9213957 TI - [The Epstein-Barr virus in patients with infectious endocarditis]. AB - Serological markers of Epstein-Barr virus (EBV) infection has been investigated in 28 patients with infectious endocarditis. In 75% of patients IgM antibodies to "early" antigen of the virus which are the marker of active viral infection occurred vs 6.2% among healthy blood donors. Specific for infectious endocarditis reaction profile (anti-EBV combination in one person) indicates reactivation of latent viral infection. The conclusion is made on the necessity of further investigation of both the role of EBV in pathogenesis of pyoseptic diseases and immunologic mechanisms for reactivation of latent viral infections. PMID- 9213958 TI - [The effect of a new hypolipemic preparation fluvastatin (Lescol) on rheological indices and hemostatic parameters]. AB - In view of inducing action of hyperlipidemia on progression of nephropathy and relationships between thrombogenesis, atherogenesis and sclerosis, the authors examined fluvastatin effects on platelet-rheological hemostasis. The 12-week course in a dose 20-40 mg/ day produced minimal side effects while its hypolipidemic action was noticeable: a 18, 21 and 20% fall in concentrations of total cholesterol, LDL cholesterol, triglycerides, respectively. The platelet rheology underwent the following changes: spontaneous platelet aggregation went down from 2.58 +/- 0.3 to 1.64 +/- 0.27 r.units, ADP-induced platelet aggregation rose from 6.5 +/- 0.66 to 8.08 +/- 0.77 r.units. No marked changes were registered in hematocrit, plasma and blood density, red cell aggregation and deformability. Thus, active lowering of blood lipids was not associated with evident inhibition of platelet activity. The absence of this feedback in lipid platelet relations probably indicates an independent significance of hemostatic disturbances in ischemic heart disease and needs further study. PMID- 9213959 TI - [The chronobiology and clinical efficacy of Daflon in the treatment of chronic venous insufficiency]. PMID- 9213960 TI - [Electrical impedance tomography in pulmonology]. AB - Application of a modern method of introscopy--electroimpedance tomography (EIT) for diagnosis of different types of lung diseases is described. The EIT system including measurement and collecting data devices, 16-electrodes array and IBM PC 486 computer was used. The results of analysis of electrotomograms have demonstrated that the EIT-system can be introduced for detecting abnormal lung fluid levels, pulmonary edema, diagnosis of lung cancer, emphysema, pleuritis, hydrothorax, sarcoidosis. The method provides high sensitivity to the changes in the body physiological state. Other advantages are: safety, fast measurements, ease of equipment transportation and maintenance, low cost. PMID- 9213961 TI - [The cerebrovascular complications of hypertension: circulatory encephalopathy and strokes]. PMID- 9213962 TI - [The current status of the diagnosis of lung cancer under the conditions of the long-term dispensary care of a population group]. AB - In 1980-1995 the authors observed 739 cases of lung cancer. A special screening program on early diagnosis of lung cancer has been developed. The centralized system of the information collection, storage and processing of all the cancer cases, follow-up of all the registered cases provided observed and corrected 5- and 10-year survival. Efficacy of screening and early treatment is shown. PMID- 9213963 TI - [The hydration of the blood and its components in patients with acute and chronic leukemias]. AB - Estimation of total, free and bound water in the whole blood, plasma and red cells of 54 patients with acute and chronic leukemia showed that patients with chronic leukemia had low levels of bound water though high levels of total and free water in the whole blood and red cells. With the progress of the disease to the terminal stage these changes become more pronounced. Plasma hydration in chronic leukemia manifested with a rise in the bound and total water, in the terminal stage bound water was reduced while free and total water increased. The above changes were not dependent on the form of chronic leukemia. Hydration of blood and its components in patients with the first attack of acute leukemia and in those in terminal stage of chronic leukemia seemed similar. PMID- 9213964 TI - [Erythremia: the activity of erythrocyte antioxidant enzymes and the association with iron deficiency]. AB - Concentration of malonic dialdehyde (MDA) and activity of antioxidant enzymes G-6 PD, glutation peroxidase (GP), glutation reductase, catalase, superoxide dismutase were measured in red cells of patients with polycythemia vera. Plasmic ions Fe3+ were estimated by means of electron-paramagnetic resonance. MDA concentration and antioxidant enzymes (except GP) in polycythemia red cells were found increased, while the activity of selenium-dependent GP was reduced, the inhibition being greatest in severe iron deficiency. It is suggested that GP activity in red cells depends on both selenium levels in the body and concentrations of non-hematic iron. PMID- 9213965 TI - [The use of cyclophosphane in the immunodepressive therapy of severe aplastic anemia]. AB - Low course doses of cyclophosphamide (1.9 +/- 0.2 g) were given to postsplenectomy patients with grave aplastic anemia (GAA). A total of 27 patients aged 14-42 years were treated, 27 control GAA patients did not receive cyclophosphamide. The cyclophosphamide therapy was controlled by T-lymphocyte functional activity assessed in active rosette formation test with levamisole in vitro. 19 (70.3%) patients were treated twice. The remission was achieved in 22 patients (81.5%), partial response was in 1 patient (3.7%), 4 patients died (14.8%). In control subjects the remission was recorded only in 4 subjects, 23 patients died 3 to 12 months after splenectomy. It is evident that low-dose courses of cyclophosphamide are highly effective in case of control over immunological activity of the pathological process. PMID- 9213966 TI - [The clinical aspects and psychopharmacotherapy of adaptive neurotic disorders]. AB - 57 neurotic patients were included in the trial because they met the following criteria: no signs of psychic disorders before emergence of psychogeny, close relation of the disease with change in social condition, only minor psychic symptoms. The patients have suffered psychic traumas caused by loss of social significance of their job, loss of financial well-being, family problems. The reactive condition was associated with psychogenic complex. The patients developed neurotic depression rather early. It consisted of 3 stages: neurasthenic disorders, anxiety, depressive condition with distress affection. Adaptative disorders need both psychotherapy and psychopharmacotherapy. Inpatients were given bensodiazepines, antidepressants, mild neuroleptics, nootropes. It is shown than the duration of the treatment course and choice of chemotherapy depend primarily on the time of seeking medical advice and start of therapeutic measures. PMID- 9213967 TI - [The psychosomatic aspects of patient adaptation to hemodialysis treatment]. PMID- 9213968 TI - [The importance of myoglobin in the pathogenesis of leptospirosis]. AB - Measurements were made of serum and urine myoglobin in 48 patients with leptospiral jaundice (LJ) and 56 patients with various acute infections. At the height of LJ blood myoglobin level reached 28.96 +/- 4.3 micrograms/l (normal concentration 0.315 +/- 0.002 microgram/l). Compared to acute pneumonia, acute viral hepatitis, tonsillitis, erysipelas, diphtheria, health values, the ratio of serum myoglobin to urine myoglobin in leptospirosis made up 45.25 against 5.4, 4.8, 6.8, 3.7, 1.8 and 1.3, respectively. A relationship was found between concentrations of myoglobin, bilirubin, creatinine in the blood and leptospirosis severity. Elevation of serum myoglobin as a manifestation of specific myositis is pathognomic for leptospirosis and contributes to the onset of acute renal failure and disturbance of bilirubin metabolism. Quantitation of blood myoglobin may be helpful as an additional test for leptospirosis severity. PMID- 9213969 TI - [Acute arterial obstruction (its etiology, diagnosis and treatment)]. PMID- 9213970 TI - [The diagnosis of eye lesions in infectious diseases]. PMID- 9213971 TI - [A case of myocardial infarct in latent porphyria cutanea tarda]. PMID- 9213972 TI - [Functional stress tests in the diagnosis of ischemic heart disease in women]. PMID- 9213973 TI - [Clinical ethics committees in Norway]. PMID- 9213974 TI - [Patients' evaluation of clinical practice and research--their subjective views tell us all]. PMID- 9213975 TI - [Suicidal behavior among young people--is it possible to prevent?]. PMID- 9213976 TI - [Cloning of animals and humans]. PMID- 9213977 TI - [Involuntary admissions in psychiatry]. PMID- 9213978 TI - [Involuntary admissions in emergency psychiatric institutions. A comparison between the county of Hedmark and the Ulleval sector in Oslo]. AB - In this paper we compare the acute psychiatric wards in Hedmark county and the catchment area of Ulleval hospital with regard to involuntary hospitalizations and admission rates for psychosis during the period 1989-94. In the former area the percentage of involuntary admissions decreased from 58% to 48% during the period, but in the Ulleval catchment area it remained fairly stable at 85%. The latter area showed a higher percentage of involuntary hospitalizations for both psychotic and non-psychotic patients. The admission rate for psychotic patients was higher in the Ulleval catchment area and tended to increase in the course of the study period. We relate our findings primarily to a higher incidence of psychiatric disorders in a city like Oslo. Structural differences in psychiatric and primary care, especially for long-term patients may also be a contributory factor. PMID- 9213979 TI - [When need is greatest--is help nearest? Help and treatment after attempted suicide among adolescents]. AB - In a comprehensive national survey among adolescents aged 14-22 years, a total of 579 (7.6% of the net sample) stated that they had at same time taken an overdose of pills or had tried to harm themselves, in some other way. The proportion of the attempted suicides who had received help or treatment in hospital or from a medical practitioner afterwards was 6%. The proportion was higher among those who had made repeated attempts of suicide and among frequent users of drugs. A total of 16% reported having received help or treatment from a psychologist or psychiatrist, and showed a higher depression score than other attempted suicides. Thirty-two percent reported receiving help from family or friends after-wards, mostly from friends. Two thirds of the boys reported not having received any help or treatment after the suicide attempt. PMID- 9213981 TI - [Consultation-liaison-psychiatry in Norway. Use of psychiatric services in somatic departmetns]. AB - The aim of the present study was to assess the use of consultation-liaison psychiatry in Norway, the resources used, the organisation and quality of the services, and whether there is a need to increase the services and improve the education. A questionnaire was mailed to all the major somatic and psychiatric departments in Norway. The questionnaires were completed and returned by 189 somatic departments (85%) and 74 psychiatric departments (55%). Each psychiatric department gives a median of two consultations each week (range 1-13). A minority of the consultations are with a specialist in psychiatry. About 60% of the psychiatric departments offer supervision of younger doctors. Psychiatric evaluation is seldom carried out when the somatic diagnosis is unclear. Most of the somatic departments are satisfied with the services. Both the somatic and psychiatric departments express a need for more extensive services and more education in medical psychology. PMID- 9213980 TI - [Are preventive measures against suicide efficient? A literature review]. AB - The increasing frequency of suicide in Norway from 1970 to 1990 has called attention. This paper raises the question of whether any controlled studies have been conducted on prevention of suicide. A search was made in Medline; the criteria for including articles were: controlled studies concerning the effects of suicide-preventive intervention by measuring the number of suicides, parasuicides or suicidal thoughts/ideation. The result of our search was 13 randomised controlled trials and two case-control studies. Most of the studies were unable to confirm any effect of suicide-preventive intervention. In most studies the experiment and control groups were far too small to arrive at significant conclusions. Prior to starting prevention programmes, it is necessary for the health service to know the potential effects. This systematic review revealed that few good controlled studies have been conducted on suicide prevention. For the time being we have to accept the suggestive evidence for certain types of intervention, and use these as a basis for action. PMID- 9213983 TI - [Achilles tendon rupture. Surgical or non-surgical treatment]. AB - Opinions differ about the proper treatment of Achilles tendon rupture. 38 patients with acute total rupture of the Achilles tendon were included in a comparative study of operative as against non-operative treatment. 21 of the patients were treated operatively and 17 non-operatively. The follow-up time was 6-53 months. Three of the non-operated patients but none of the operated group experienced major complications. Ten of the non-operated patients and 14 of the non-operated group experienced minor complications. In the non-operated patients the plantar-flexion range was significantly reduced in the injured foot compared with the other foot (p = 0.03). Because of more re-ruptures and reduced muscle strength in the non-operative group, operative treatment is recommended for active persons. Non-operative treatment may be considered for older people. PMID- 9213982 TI - [Integrated psychiatric care planning]. AB - In order to design an integrated plan for services, all long-term psychiatric patients in Trondheim were registered by professionals on both municipality and county level. Data were obtained from interviews with the patients, supplemented by information from professional personnel. Individual plans, including type and amount of services needed, were prepared for each patient anonymously, to form the basis of a mutually beneficial integrated plan for psychiatric services. A total of 507 patients were registered (0.5% of the population over 18 years). Studying individual patients and their needs makes it possible to make optimal use of limited resources. The sharing of knowledge improves cooperation and coordination, to the benefit of a neglected group of patients. PMID- 9213984 TI - [Torsion of the spermatic cord. A clinical material on 109 patients]. AB - The authors review experiences from the operation of 109 patients for torsion of the spermatic cord at a single surgical clinic. The peak incidence was seen amongst adolescents. The patients reported the sudden onset of severe scrotal pain. Clinical examination showed a tender scrotal mass. About half of the patients showed testicular retraction or scrotal erythema. 22% underwent orchidectomy because of gangrene. All these patients had a history of more than 24 hours. On the other hand, 95% of patients with a vital testis had a history of less than 24 hours. Significant testicular injury will occur in a large share of these patients as well. The true urgency of this condition is emphasized. PMID- 9213985 TI - [Occupational injuries and prolonged sick listings during the period 1989-93]. AB - Norway lacks reliable data on the magnitude and development of occupational injuries. Each year about 25,000 occupational injuries and 2,000 occupational diseases are reported to the Labour Inspection. These constitute only one-third of all expected injuries and accidents related to the work environment. The aim of this study was to estimate the magnitude of occupational injuries in Norway causing long-term certification of illness. There was a 35% reduction in the incidence of occupational injuries in Norway from 1990 to 1993. The reduction applies to men and women combined, all age groups and most counties. The distribution of characteristics by age, sex, diagnosis and place of residence corresponds well with previous, limited studies. Injuries, especially fractures, account for approximately 70 per cent of the occupational injuries. Possible explanations for the decrease in incidence could be preventive efforts, a tighter labour market, a change from primary and secondary to tertiary industry, and the overall reduction in long-term certification of illness in Norway from 1990 to 1993. PMID- 9213986 TI - [Clinical ethics committees are tested in Norway]. AB - Health care ethics committees are now established at three hospitals in Norway as a result of a three year project in cooperation with the Centre for Medical Ethics and the Ministry of Health and Social Affairs. This article describes the project and discusses various difficulties regarding the establishment and functioning of health care ethics committees, such as their mandate and membership, the understanding of ethics and methodology, the role of ethical expertise and the relation between physicians' decision-making responsibility and a health care ethics committee. PMID- 9213987 TI - [The value of autopsy in current health care. An analysis of autopsies performed during one year at the Regionsykehuset in Trondheim]. AB - We have reviewed the reports from 222 autopsies performed during one year (1994) at our Department of Pathology. The majority (74%) of all autopsies were performed on patients from the Department of Internal medicine at our hospital. Autopsy findings were correlated with the clinically suspected causes of death, as revealed by the death certificates and the clinical information in the written requests for autopsies. The direct cause of death, as found at autopsy, had been suspected by the clinicians in 70% of cases. The underlying cause of death, as found by the pathologists at autopsy, had been diagnosed by the clinicians in 75% of the cases. In 26 cases (12%) the underlying cause of death had neither been diagnosed nor suspected by the clinicians. In the last mentioned group, 77% of the patients were more than 70 years of age. In six cases where the underlying cause of death had not been diagnosed or suspected by the clinicians, the findings at autopsy were classified as type-1 findings. These are findings, if they had been known when the patient was still alive would probably have led to therapy that night have improved the prognosis. PMID- 9213988 TI - [Practical problems associated with long-term opioid therapy]. AB - Long-term opioid therapy results in physical dependency. Therefore abrupt cessation of opioid treatment may cause acute abstinence symptoms. The doses of opioids should always be reduced gradually, even when other treatment options give good pain relief. Patients receiving long-term treatment with opioids become more sensitive to the opioid antagonist naloxone. Inappropriate use of naloxone in these patients can also result in acute abstinence symptoms. Opioid treatment is limited by toxic side effects, and the problem can be solved by changing the opioid. The author describes three cases to illustrate these problems. PMID- 9213990 TI - [Epidemiology and public health--useful tool or a problem?]. PMID- 9213989 TI - [Assessment of one's own functional status. COOP-WONCA questionnaire charts in clinical practice and research]. AB - Primary health care has needed an instrument that can be used to assess patients functional health status. Since 1988 the Classification Committee of WONCA, the World Organisation of Family Doctors, has been working on a revised edition of the Dartmouth COOP Functional Status Assessment Charts. The six COOP-WONCA Charts are described: physical fitness, feelings, daily activities, social activities, change in health and overall health. The charts have been tested against many other instruments and the validity has been found to be good. A Norwegian test retest reliability study, the first on the revised COOP-WONCA Charts, showed good reliability. For 35 out of 40 of the office patients included in the study, either the answers were the same at test-retest, or "change of health" was consistent with changes in other variables. A study in a group of 171 asthma patients showed unchanged mean scores after six months. The results from a population study in the municipality of Ullensaker, comprising 2,864 persons, are presented as a basis for comparison. The COOP-WONCA instrument is simple and easy to use and understand. It meets the requirements for a useful tool in clinical practice and research and seems to provide valuable, additional information in daily practice. The COOP-WONCA Charts represent a simple and good method for assessing functional status. PMID- 9213991 TI - [Suicide prevention--professional choices, one what basis?]. PMID- 9213993 TI - [A report on the mental status in the land "down under"]. PMID- 9213992 TI - [Go out and make all the people to be my patients]. PMID- 9213994 TI - [Prevention of risks]. PMID- 9213995 TI - [Prevention of fractures in the elderly]. PMID- 9213996 TI - [Alcohol and public health]. PMID- 9213997 TI - [Scholarship holders at the medical faculty in Oslo]. PMID- 9213998 TI - [Research and development of the Norwegian drug industry in 1995]. PMID- 9214000 TI - [Honour for the veterinarians]. PMID- 9213999 TI - [Tough practice in 1904]. PMID- 9214001 TI - [All physicians should concern on prevention!]. PMID- 9214002 TI - [Twisted neck--torticollis. Efficient treatment is available]. PMID- 9214003 TI - [Subarachnoid hemorrhage is still a very serious disease]. PMID- 9214004 TI - [Subarachnoid hemorrhage in Vestfold county. Occurrence and prognosis]. AB - This prospective study comprises all patients treated for spontaneous non traumatic subarachnoid haemorrhage between May 1991 and December 1995 in the county of Vestfold, Norway. A total of 76 patients was recorded, giving an incidence of 8.1 per 100 000 per year. Mean age at time of bleeding was 52.7 years. In 36 patients the cause of the bleeding was an intracranial aneurysm; most of the aneurysms were localized to the anterior communicating artery and middle cerebral artery. In seven patients the cause was arteriovenous malformation. 23 patients (30%) died because of their subarachnoid haemorrhage; 15 from the primary bleeding and eight because of re-bleeding. The mortality for patients aged over 60 years was 48%, and for patients younger than 60 years 19%. There was a strict correlation between the initial clinical condition (Hunt & Hess scale) and the final outcome (Glasgow Outcome Scale). PMID- 9214005 TI - [Perimesencephalic subarachnoid hemorrhage]. AB - Acute, non-traumatic subarachnoid haemorrhage is usually caused by a ruptured aneurysm. This is a serious condition involving high mortality. Perimesencephalic haemorrhage has recently been identified as a clinical subset of subarachnoid haemorrhage. A pattern of haemorrhage predominantly limited to perimesencephalic cisterns on CT is highly predictive of a normal angiogram, and is connected with an excellent prognosis. We describe two patients with perimesencephalic haemorrhage. PMID- 9214006 TI - [Terson's syndrome. Subarachnoid hemorrhage combined with vitreous body hemorrhage]. AB - Terson's syndrome consists of the combination of acute intracranial haemorrhage (most often an aneurysmal subarachnoidal haemorrhage) and profuse bleeding in the vitreous body of one or both eyes. The incidence is from 2% to 16% of patients with subarachnoidal haemorrhage. These patients are often confused in the early phase of their disease, and their decreased visual acuity is often not recognized. We describe findings in a patient recently seen in our department, with subarachnoidal haemorrhage and gross bleeding in the vitreous body of both eyes. We emphasize the importance of being aware of the syndrome, both in order to provide adequate nursing care, and to be able to perform early vitrectomy to restore visual function. PMID- 9214007 TI - [Treatment of spasmotic torticollis with botulinum toxin A. A 5-year experience]. AB - 60 patients have been treated with botulinumtoxin A for cervical dystonia since december 1990. These patients have been seen at 472 visits and received at least two treatments. 55 (92%) patients noted improvement, and marked improvement was noted by 50 out of 60 patients (83%). Further improvement after repeated treatments has been seen up to five years. Only one patient experienced no effect after several treatments with marked improvement. This might have been due to production of antibodies against the toxin. Side effects occurred after 9% of the toxin injections, and dysphagia was the symptom most often reported. The side effects were usually mild and transient, and never gave reason to terminate the treatment. PMID- 9214008 TI - [5-year experience with a clinic for amyotrophic lateral sclerosis]. AB - An out-patient service for patients suffering from amyotrophic lateral sclerosis (ALS), the ALS-clinic, was established at the Department of Neurology, Haukeland Hospital, in 1990. The number of ALS patients who were hospitalised during the period 1990-1995 was 59, with a mean stay in hospital of 14.8 days. Eleven of the patients died in hospital. The ALS-clinic had 127 consultations during the same period, with a mean of 2.2 consultations per patient. Speech difficulties were the dominating problem at 26 of the consultations. 32 patients experienced feeding difficulties, and a percutaneous endoscopic gastrostomy was performed in nine cases. Respiratory problems dominated in ten patients, but only two of these patients wanted a home ventilator. Various assistive devices were adapted for 16 patients. PMID- 9214009 TI - [Congenital dyserythropoietic anemia type III. A case report]. AB - Congenital dyserythropoietic anaemia type III is a rare disorder characterized by mild to moderate anaemia, ineffective erythropoiesis, and morphologic abnormalities of mature red blood cells and their precursors. The most extraordinary feature of this condition is the large number of multinuclear erythroblasts found in the bone marrow, some containing up to 12 nuclei. This type of anaemia is an autosomal dominant inherited disorder, though sporadic cases have been described. Little conclusive is known about the pathogenesis of congenital dyserthropoietic anaemia type III. At present the management of patients consists of observation and supportive care. We describe a 20 year old man who was admitted to a county hospital, showing the typical features of this rare illness. He had a Hb value of 10.2 g/100 ml. PMID- 9214010 TI - [Splenic atrophy and fatal pneumococcal infection in inflammatory bowel disease]. AB - Fatal pneumococcal disease occurred in a young woman with ulcerative colitis that had been quiescent for ten years. Howell-Jolly bodies were present in the peripheral blood smear. Severe atrophy of the spleen (weight 7 g) was discovered at autopsy. The evidence that hyposplenism is associated with inflammatory bowel disease is reviewed and it is suggested that these patients should undergo ultrasound examination and examination of blood smear for detection of functional hyposplenism. We conclude that pneumococcal vaccination should be considered for all patients with inflammatory bowel disease. PMID- 9214011 TI - [Central nervous pain in patients with spinal cord injury. Medical and surgical treatment]. AB - About 50% of patients with spinal cord injury suffer from persistent central neurogenic pain. The authors review the case of a patient with traumatic paraplegia who developed persistent central neurogenic pain. The pain was described as burning in the buttock area, icing in the rectum area and as lancinating pain to the lower extremities. The combination of amitryptilin and morphine had a slight, short-term effect, but the pain did not respond to treatment with simple analgetica, dextropropoxyphen or ketobemidone, neither administered alone nor in combination with tricyclic antidepressants, carbamazepine or baclophen. Transcutanous nerve stimulation and acupuncture had no effect. The patient was operated on by means of the computer-assisted dorsal root entry zone (DREZ)-microcoagulation technique 2.5 years after the trauma. This technique is described in brief. The prevalence and classification of neurogenic pain, and possible medical and surgical treatment, are also discussed. PMID- 9214012 TI - [Splenic abscess. Diagnosis and treatment]. AB - A diagnosis of hospital discharges shows splenic abscess to be a rare condition, with one case per 10,000 discharges. Haematogenous seeding to the spleen from an infection at a distant site, most often endocarditis, has been the most common predisposing condition but an increase has been observed in immuno-suppressed patients too. Fever, leukocytosis and left upper quadrant pain are suggestive, but the signs and symptoms of splenic abscesses are often non-specific. Ultrasound and computed tomography are reliable diagnostic tools. Splenectomy and antibiotics have been the treatments of choice, with increasing use of percutaneous drainage as an alternative, in order to preserve splenic function. We describe a patient with a salmonella splenic abscess that was treated with percutaneous drainage and ciprofloxacin. PMID- 9214013 TI - [Long-term result of a life style intervention group program]. AB - 156 former participants in a lifestyle modification programme for persons with coronary heart disease, or at high risk of developing this disease, were invited to a follow-up examination. The aim was to evaluate the long-term effect of a lifestyle intervention programme. Median time since completion of the programme was 3 years. The participants' serum cholesterol level at the time of the control examination was lower than before they attended the programme, but slightly higher than at the end of it. The body weight was similar at the control and before participation. The self-reported amount of physical activity of persons with established coronary heart disease was higher at the control examination than before the programme. For persons with no established coronary heart disease, the self-reported level of physical activity was the same before and after attending the programme. The most pronounced effect as regards a reduction of the risk factor profile was seen among the participants with established coronary heart disease. The number of smokers was reduced, but not significantly. PMID- 9214015 TI - [Rehabilitation after stroke. Programs at the Sunnaas hospital]. AB - The article describes the rehabilitation programmes for stroke patients at Sunnaas Hospital, a Norwegian university hospital for rehabilitation. Patients with complex and less frequent problems following a cerebrovascular accident can attend this third line hospital for primary or secondary rehabilitation. In addition, special short-term programmes are offered to assess the potentials for rehabilitation, competence for a driver's licence, vocational ability and swallowing problems, as well as to judge the need for technical aids for communication, mobility and management of the surroundings. The various programmes are described in detail. During 1996 a total of 306 stroke patients, median age 57 years, attended the hospital. The hospital aims to continue to be a highly specialized centre for stroke patients, also in the future, with different kinds of comprehensive and multidisciplinary rehabilitation programmes for these patients. PMID- 9214014 TI - [Integration of preventive medicine in an emergency department. A pedagogic model for improvement of communication]. AB - At the Medical Department, Telemark Central Hospital, a project has been going on for five years now to evaluate consultations in lifestyle groups in preference to individual consultations for persons with dyslipidemia. 363 persons were recruited to participate in a series of 5 group consultations at intervals of 3 months, each session to last for 2 hours. Altogether 1469 consultations were of this type. After the first session 79% said they preferred lifestyle group consultations, rather than spending their share of the allotted time on personal consultations, and after the fifth session 81%. The concept has been extended to include patients with chronic disease (asthma and chronic inflammatory bowel disease), with the principal aim of improving the patients' understanding of their disease, and showing them how to control it themselves. The project has attracted much attention, and a consultant in preventive medicine has recently been appointed to the staff. We think it important in terms of impact that the initiative to establish local expertise in preventive medicine emerged from a department that deals with emergency admissions due to lifestyle-related diseases. PMID- 9214016 TI - [How do antioxidants work?]. AB - Reactive oxygen species, formed by incomplete reduction of molecular oxygen, may cause oxidative stress in the organism and be involved in the pathogenesis of many diseases. A number of anti-oxidants are necessary to counteract the harmful effects. Some antioxidants are enzymes which catalyze the breakdown of reactive oxygen species, some act by chelating transition metals, which makes them non reactive, while chain-breaking antioxidants act by halting the cascade of free radical reactions. The effect of redox active antioxidants depends on their redox status. In some circumstances antioxidants can have pro-oxidant effects. The mechanisms may be reductive release of metals, accumulation of the oxidized form of redox active antioxidants, or merely an unfavourable equilibrium between various antioxidants. At present there is no basis for recommending the prophylactic use of commercially produced antioxidants. In particular, the optimal combinations of antioxidants are not yet known. If the aim is to improve the antioxidant defence of the body, our advice is still a mixed diet and regular physical activity. PMID- 9214017 TI - [Preventive and health promotion work in family practice. Dilemmas and proposals on general guidelines]. PMID- 9214018 TI - [Physician's personal responsibility. Experiences from the Diprivan issue]. PMID- 9214019 TI - [Iron and pregnant women--guidelines on the wrong road?]. PMID- 9214020 TI - [No, there is no magic grip]. PMID- 9214021 TI - [Not smoking-free, but better than in Norway!]. PMID- 9214022 TI - [Service onboard MV Anastasis--an inspiring professional challenge]. PMID- 9214023 TI - [Diagnostic criteria for acute myocardial infarction]. PMID- 9214024 TI - [Blue lights and sirens in emergency services]. PMID- 9214025 TI - [To consult a physician]. PMID- 9214026 TI - [The electronic record--little bit hera--and little bit there?]. PMID- 9214027 TI - [Mutual agreements for use of scientific databases in health services?]. PMID- 9214028 TI - [Clinical laboratory research on the properties of Aquafresh toothpaste]. AB - The study of the teeth in 35 patients with chronic catarrhal generalized gingivitis and periodontitis of moderate severity using tooth paste Aquafresh for a months confirmed high cleansing, antiinflammatory and refreshing potential of the paste which, moreover, has a good taste and mint flavor. Laboratory tests showed low abrasibility of the paste, its ability to raise by 12.5% the resistance of the enamel to demineralizing action of the lemon juice. Not a single case of the paste-induced irritation of the oral mucosa and teeth was registered. PMID- 9214029 TI - [The effect of a whitening gel containing carbamide peroxide on enamel and dentin ultrastructure]. PMID- 9214030 TI - [A new generation of automated laser surgical apparatus with computer control]. PMID- 9214031 TI - [Antiseptics in the combined treatment of patients with perimaxillary abscesses and phlegmons]. AB - The efficacy of local use of antiseptic solutions (chlorhexidine bigluconate, chlorfillipt and furaciline) on clinical presentation and microflora of purulent wounds was studied in 236 patients with maxillofacial abscesses and phlegmons. The wound microflora was represented by a wide spectrum of opportunistic agents. A pronounced therapeutic effect was achieved in patients with staphylococcal infection. Marked resistance to the drugs was noted in alpha-hemolytic streptococcus. PMID- 9214032 TI - [The diagnosis and treatment of contact odontogenic mediastinitis]. AB - Contact odontogenic mediastinitis is a complication of advanced phlegmons of the face and neck. 22-year experience in the treatment of 107 patients with odontogenic mediastinitis, clinical and laboratory criteria of its diagnosis and differential diagnosis with advanced phlegmons are presented. The programs of intensive therapy adjusted for the disease phase and surgical treatment contributed to lowering of lethality from 50 to 15.3%. PMID- 9214033 TI - [Surgical treatment methods in combined nasal deformities]. AB - The paper presents the procedure of surgical correction of the twisted nose, with high humped ridge of the nose, in particular. Inferior resection of the segment from the cartilage of the nasal septum and cross-lateral osteotomy are followed by manual impression of the ridge. Mobilized bone clivi are pushed inside, osteochondral part of nasal pyramid moves backwards under the base of the maxillary frontal processes. The ridge of the nose moves backward till the frontal septum reaches the vomer. The wedged pyramid does not allow lateral dislocation of the ridge. Anteroinferior part of the septum is fixed by chrome suture to spina nasalis tissues. The procedure is superior to N. M. Mikhel'son's method because it provides more stable cosmetic results and makes it possible to correct humps of the nasal ridge. PMID- 9214034 TI - [The one-stage replacement of single anterior defects of the dental arches with endosseous implants using light-hardening filling materials]. AB - Implantation with one-stage repair of a tooth crown using helioprogram has been conducted in 42 patients with single frontal defects of the teeth. A 2.5-3-year follow-up enables the conclusion on this method's safety, good functional and cosmetic results. Thus, it is worth introduction into wide practice. PMID- 9214035 TI - [The status of the oral cavity in children with defects in the central nervous system and locomotor apparatus of congenital and hereditary origin (infantile cerebral palsy, spinal cord hernias, myopathies)]. PMID- 9214037 TI - [Nonspecific lymphadenitis and adenophlegmon of the maxillofacial area and neck in children]. AB - The study into the causes underlying nonspecific lymphadenitis (NSL) and adenophlegmons of the maxillofacial region and the neck (AF) revealed a great variety of etiological factors and primary infectious foci in these diseases. Of 204 children admitted to hospital of the city of Derbent, odontogenic genesis of the disease was determined in 27.45% of cases; dermatogenic, stomatogenic in 23.04 and 12.74% of cases, respectively. ENT and systemic diseases were responsible for NSL and AF in 13.23 and 3.43% of cases. The cause remained unclear in 20.09% of patients. NSL and AF occurred most frequently in the coldest (January, February) and the hottest (July, August) months of the year. PMID- 9214036 TI - [The structural characteristics of the bony tissue at the surface of an implant with hydroxyapatite spray-coated with excimer and CO2 lasers]. AB - Rat experiments were performed to study osteogenesis and osseointegration in implanting fragments of dental titanic implants into the spongy bone. The implant spray-coating with hydroxyapatite produced by excimer and CO2 lasers stimulates osteogenesis. Bone tissue integration with implant proceeds more actively in response to CO2 laser radiation. The weakest integration was registered between the metal and bone tissues. PMID- 9214038 TI - [The characteristics of the clinical course of inflammatory diseases of the maxillofacial area in children]. AB - As shown by investigations in 1990-1994, the number of cases with maxillofacial inflammation in children tended to an increase. Etiology became primarily odontogenic (69.5% of patients). Inflammatory processes developed against aggravated premorbid backgrounds. The group of risk was children aged 7-12 years. Yearly diagnosis, valid and adequate treatment combining immunostimulants, physiotherapeutic procedures, laser radiation reduced the time of rehabilitation and treatment, stimulated body response. Such policy proved effective in management of maxillofacial inflammation. PMID- 9214039 TI - [The forms of property and the organizational legal forms of juridical persons in dentistry]. PMID- 9214040 TI - [A case of a gunshot wound of the face by a fireworks rocket]. PMID- 9214041 TI - [Osteoplasty in mandibular defects]. PMID- 9214042 TI - [A clinical evaluation of fillings made from Iartodent composite filling material]. PMID- 9214043 TI - [The restoration of metal-ceramic dentures in the patient's mouth]. PMID- 9214044 TI - [The outlook for scientific research in the field of therapeutic dentistry]. PMID- 9214046 TI - [Scientifically based medical practice]. PMID- 9214045 TI - [Peptic ulcer and ulcer dyspepsia]. PMID- 9214047 TI - [Ulcer, personality and stress]. PMID- 9214048 TI - [Measurement of gastric acid secretion and pH]. PMID- 9214049 TI - [Language politics in health care]. PMID- 9214050 TI - [Nutrition in hospitals]. PMID- 9214051 TI - [Helicobacter pylori in 1997]. AB - In this review Helicobacter pylori (H. pylori) infection and its relation to different diseases is presented. H. pylori doesn't cause inconvenience to most infected people, though all infected persons have chronic active gastritis. The 10 year risk of peptic ulcer for people infected with H. pylori is about 10%. Randomized double-blinded trials have shown that eradication of H. pylori can cure most patients with peptic ulcer disease. Some people infected with H. pylori develop atrophic gastritis which is a risk factor for development of gastric cancer. It is not known if H. pylori screening and eradication would have a prophylactic effect against gastric cancer. It is also unknown if persons with non-organic dyspepsia and persons in long-term treatment with proton-pump inhibitors would benefit from H. pylori eradication. PMID- 9214052 TI - [The influence of adhesion molecules in inflammatory bowel diseases]. AB - Cell adhesion molecules are a group of membrane-bound molecules that are involved in the binding of circulating leukocytes to activated endothelial cells and in the migration of these leukocytes towards the inflamed area. Cellular adhesion molecules are divided into four major families: selectins, immunoglobulin-like cell adhesion molecules, integrins, and carbohydrate ligands. Selectins (E-, P-, and L-) and their ligands, ICAM-1, VCAM-1, VLA-4, beta 2 integrins, and presumably also PECAM-1, MAdCAM-1, and alpha 4 beta 2 have been shown to be central in the development of the anatomic lesions in ulcerative colitis and Crohn's disease. Even though investigations on the effect of antibodies against cellular adhesion molecules in relation to inflammatory bowel disease are limited, an anti-inflammatory effect may be possible. Drugs which interfere with the function of cellular adhesion molecules should be tested in trials, as it only by this way can be determined whether they are beneficial and appropriate. At present, however, it is known that glucocorticoids and 5-aminosalicylic acid, which are the cornerstones in inflammatory bowel disease treatment, both interact with the synthesis and function of cellular adhesion molecules. PMID- 9214053 TI - [Dyspepsia in general practice. A study among users of anti-ulcer agents]. AB - Five doctors in general practice sent identical questionnaires to all those of their listed patients who in the previous year had received one or more prescriptions for an ulcer medication in the groups H2 receptor antagonists, acid pump inhibitors, bismuth and sucralfate preparations. The patients were identified by the pharmacy. Two hundred and fifty-nine questionnaires were returned, and the response rate was 83%. Four point nine percent of an adult population of 7160 received in the year 1993 at least one prescription for ulcer medicine, and 0.87% were in constant treatment. On average the doctors met 49 dyspeptic patients per 1000 adults per year. This is far more than found by gastroenterologists in studies of different ways of handling the dyspeptic patient in general practice. PMID- 9214054 TI - [General practitioners' handling of patients with dyspepsia]. AB - The handling of patients with dyspepsia by 102 general practitioners was investigated prospectively using a standardised application form for gastroscopy followed by a questionnaire to the practitioners. A mean of 5.3 indications were stated among 16 performed referral possibilities in 143 applications. Referral because of long-standing symptoms, ineffective treatment trial and atypical dyspepsia were more often stated than certain important risk factors (high age, NSAID-ingestion etc.). In 23% of cases no specific suspicion of organic disease was stated. The questionnaire was answered by 85%. More than 90% of doctors would use a treatment trial before further investigation of dyspeptic patients with first time consultation. A wide variation of medical empirical treatment was indicated. The practitioners wished to retain the open access model for referring, but a great majority desired practical guidelines for the handling of dyspeptic patients. This study confirms the relevance of such guidelines to improve present practice. PMID- 9214056 TI - [NSAID and ulcer complications. An analysis of risk factors]. AB - Use of NSAIDs is recognized as an important cause of peptic ulcer complications. The aim of this nested case-control study was to identify risk factors for NSAID related ulcer complications. Cases were consecutive NSAID users admitted with an ulcer complication (n = 94), and controls were a random sample of all NSAID users without ulcer complication identified by a pharmaco-epidemiological database (n = 324). Risk factors for patients at start of NSAID-therapy were: high age: 60-75 yr; Odds Ratio (OR) 3.5 (95% CI: 1.8-7.0); > 75 yr: OR 8.8 (4.3-18.1); male sex: OR 1.7 (1.0-3.0); ulcer history: OR 2.5 (1.2-5.1); steroid treatment: OR 2.0 (0.8 4.6); smoking: OR 1.6 (0.9-2.7); alcohol use: OR 1.8 (0.9-3.6). Risk factors for patients on NSAID-therapy were: high age, male sex, ulcer history, and smoking, and furthermore dyspepsia: OR 2.1 (1.0-4.2), especially NSAID-related dyspepsia: OR 8.9 (4.1-19.2). Risk was lower for patients treated more than three months. In conclusion, risk measured from this design can be shown to correlate strongly with the rate difference, a measure that is more clinically relevant than conventional relative risk estimates. Strong risk factors for NSAID-related ulcer complication are high age, male sex, ulcer history, and dyspepsia related to the NSAID therapy. PMID- 9214055 TI - [Alcohol intake and risk of liver disease--significance of gender. A population study]. AB - The association between self-reported alcohol intake and the risk of future liver disease was studied in a population-based prospective cohort of 13,285 men and women aged 30-79 years. Diagnoses indicating alcoholic liver disease (n = 261) or cirrhosis (n = 124) were obtained from the Danish National Health Registers. The cumulated observation time was 130,558 person-years. A dose-dependent increase in risk of developing liver disease was observed with increasing alcohol intake, with the steepest increase among women. At an alcohol intake of 7-13 beverages per week for women and 14-27 beverages per week for men the relative risk was significantly greater than 1. Women had a significantly higher relative risk of developing alcohol related liver disease than men for any given level of alcohol intake. In the general population, self-reported current alcohol intake is a good predictor of the future risk of alcoholic liver disease. PMID- 9214057 TI - [Inflammatory pseudotumor--differential diagnosis in lung tumor in a child]. AB - A case of a twelve-year-old girl with a big solid tumour in the right lung is presented. As malignancy could not be excluded, she was operated. A lobectomy was performed. Frozen section showed no malignancy. Histology showed inflammatory pseudotumour, which is a rare but important benign lung tumour in childhood. The recommended treatment is conservative resection. PMID- 9214058 TI - [Point, crowns and pennies]. PMID- 9214059 TI - [Bell's palsy treated with acyclovir]. PMID- 9214060 TI - [Phenemal to pregnant women and children's intelligence and quality of life 31-33 years later]. PMID- 9214061 TI - [The new prevention during pregnancy]. PMID- 9214062 TI - [Male infertility]. PMID- 9214063 TI - [Syndrome X. Angina pectoris in patients with angiographically normal coronary arteries]. PMID- 9214064 TI - [An oncologist on the Internet]. PMID- 9214065 TI - [Borna disease virus. An etiological agent in neuropsychiatric diseases?]. AB - Borna disease virus has long been recognized as a cause of sporadic cases and epidemics of meningoencephalomyelitis in horses and sheep in southern parts of Germany. however, sero-epidemiological surveillances indicate that Borna disease virus has a global distribution in horses, without the recognition of clinical manifestations associated with the infection, in other parts of the world. During the past five years evidence has been presented suggesting that humans also can become infected with this virus or a closely related virus. A significantly increased sero-prevalence is seen in patient populations with a variety of neuropsychiatric and behavioral disorders, suggesting that Borna disease virus or a closely related virus may play an etiological role in some of these diseases. A review of the literature is given. PMID- 9214066 TI - [Hemoglobinopathy in the county of Copenhagen]. AB - In Copenhagen County, haemoglobinopathy was centralized to the Department of Haematology L, Herlev University Hospital from January 1995. All pregnant women of relevant ethnic origin admitted to the obstetric departments of the Country in 1995, were examined by haemoglobin electrophoresis. furthermore, we performed haemoglobin electrophoresis on immigrants admitted on suspicion of haemoglobinopathy. 24 (4.8%) of 505 examinations in pregnant women were abnormal. 12 reflected a carrier condition for either beta-thalassaemia or sickle cell disease; 53 of 82 examinations in non-pregnant patients were abnormal; 29 had beta-thalassaemia minor and the rest included the haemoglobin variants C, D, E, H and S, mostly in a heterozygous from. The genetic lesions, all of which were mutations, were characterized by molecular genetic analysis in 13 cases with demonstrated beta-gene disorder. The gene frequency of haemoglobinopathies among immigrants to Denmark is common. The Danish health care system must therefore be prepared to address this problem including the clinical aspects, screening and molecular biological examinations, prenatal diagnosis and genetic counselling. PMID- 9214067 TI - [The health project Ebeltoft: health check ups and discussions in general practice. Basic data from a 5-year, prospective, randomized, controlled population study]. AB - A study was carried out to investigate people's interest in participating in health check-up and in discussions about health with their own general practitioner, participants' health status, the proportion who received health advice following health check-up, and the lifestyle goals they set following discussion with their general practitioner. This study reports the baseline data from a five-year randomized, controlled, prospective, population-based study in general practices in Ebeltoft, Denmark. All general practitioners from the four practices in Ebeltoft and a random sample of 2,000 people aged between 30 and 50 years were invited to participate. Participants were randomly divided into three groups-one control group and two intervention groups. One intervention group was given a health check-up which included a range og tests (Table 2 and 3); this group received written feedback from the general practitioner. The other intervention group was also given a health check-up and written feedback, in addition, they were given the opportunity to attend their general practitioner to discuss health-promoting measures. A total of 1370 people participated in the study (69% response rate). Health advice was given to 76% of 905 participants following health check-up. Almost all of the 456 participants (96%) who were offered the opportunity of discussing their health with their general practitioner took up the offer: 64% of the 456 participants reported that they had decided to undertake lifestyle changes. Eleven of those who discussed their health with the doctor were referred to a specialist (2%). There was considerable interest in participating in health promotion. Three out of four of those who had a health check-up were given health advice. Two out of three of those who were offered a health talk with the general practitioner appeared willing to make relevant lifestyle changes. Longterm follow up is needed to determine effects and side effects of health check-up and health talks. PMID- 9214068 TI - [Patients' evaluation of coercion in a psychiatric department. Interview studies in psychiatric departments for adult patients at the Hillerod hospital]. AB - During an 11 month period, use of coercion was prospectively registered in a psychiatric department. Of all patients who had been subjected to coercion 36% were (n = 86) interviewed subsequently. Eleven percent of the interviewed patients did not know, that they had been submitted to coercion, 22% did not know the reason and 30% did not agree with the motivation for the decision. Forty seven percent were satisfied with the information they had been given concerning how to complain of their treatment, 50% were satisfied with their adviser and 70% were satisfied with their overall admission. Sixty-five percent accepted that there should be a law allowing the use of coercion in psychiatry. PMID- 9214069 TI - [Prognosis after AMI--are there gender differences?]. AB - To examine the prevailing hypothesis that females fare worse than males after acute myocardial infarction, we compared short-term (15 days) and long-term (ten year) prognosis after acute myocardial infarction for the two sexes. Three thousand and seventy-three consecutive patients with acute myocardial infarction were followed for 10 years after a first registration in the Danish Verapamil Infarction Trial database in 1979-81. Early mortality increased significantly with age (p < 0.0001), but was not significantly related to sex, with a 15 days mortality of 17% in females and 16% in males. Ten year mortality in patients alive day 15 was 58.8%. Hazard ratio for females versus males after adjustment for age was 0.90 (0.80-1.01). Ten year reinfarction rate was 48.8% with age adjusted hazard ratio for females versus males of 0.90 (0.78-1.04) and ten year mortality after reinfarction was 82.3%, with age adjusted hazard ratio in females versus males of 0.98 (0.82-1.16). No difference in cause of death was found between the two sexes. We conclude that sex by itself is not a risk indicator after acute myocardial infarction. PMID- 9214070 TI - [Volumes of synovial membrane and joint effusion determined by MR imaging. Markers of severity and/or activity in rheumatoid arthritis?]. AB - Volumes of synovial membrane and joint effusion were determined by MRI in 36 knees with gonarthritis and five healthy knees. In 18 knees MRI was performed before and immediately after arthrocentesis. The difference between MRI determined and syringe-determined volumes of aspirated joint fluid was 0-7 ml, median 2 ml, corresponding to 0-18%, median 7%, of the pre-aspiration effusion volume. Synovial membrane volumes, determined before and after arthrocentesis varied 0-10 ml, median 3 ml (0-17%, median 7%). No significant systematic misinterpretation of the borderline between joint fluid and synovium was found. The interobserver variation was < 15% in all measurements of synovial volumes > 10 ml and effusion volumes > 5 ml. Patient repositioning resulted in variations < 10%. The synovial volumes in clinically active, clinically inactive and healthy knees were statistically significantly different (median 79 ml, 21 ml and 3 ml, respectively). We conclude that effusion volumes, and in all probability also synovial membrane volumes, can be determined by MRI with a maximal error of approximately 20%. The synovial volume is related to the inflammatory activity of the joint. The results encourage further studies of the value of effusion and synovial membrane volumes as markers of the activity and/or severity of joint inflammation. PMID- 9214071 TI - [Testicular carcinoma in situ detected by ultrasound in infertile men]. AB - The findings of non-palpable testicular tumour and contralateral carcinoma in situ testis (CIS) in two men, examined because of infertility, are described. Both patients had no symptoms other than infertility and bilateral testicular atrophy. Ultrasonography showed tumour in both cases and abnormal ultrasonic texture of testicular tissue in the contralateral testis. Both patients had seminomas and contralateral CIS. These cases show the value of ultrasonic examination of testis in men examined for infertility, and ultrasonic examination is recommended in these men, particularly if they have testicular atrophy. PMID- 9214072 TI - [Acute organophosphate poisoning after spraying against dog fleas]. AB - A case of moderate organophosphate intoxication in a 49 year-old male who sprayed against dog-fleas in his home is presented. The patient had symptoms of cholinergic excess, presenting as gastrointestinal hyperactivity, salivation, sweating, muscle fasciculations and confusion after in-door spraying with windows and doors kept closed. He recovered after 4 mg of atropine. S-cholinesterase was 3900 U/l (normal range 3800-10,000 U/l) measured with an activity assay, and increased to 6700 U/l the following day. PMID- 9214073 TI - [Doxycycline induced photoonycholysis]. AB - A case of phototoxic onycholysis secondary to treatment with doxycycline is presented. Tetracyclines are known to provoke photosensitivity reactions in some individuals exposed to sunlight. The typical photo-toxic manifestation is erythema, but cases with onycholysis have also been reported in the past. With travel to regions with sunny climates it will be important to be aware of the risk of developing onycholysis following the use of tetracyclines and to avoid the most potential phototoxic derivates, especially doxycycline. PMID- 9214074 TI - [Adjuvant tamoxifen treatment in breast cancer: is 5-year treatment sufficient?]. PMID- 9214075 TI - [Prevention of hepatitis B]. PMID- 9214076 TI - [Is the prevention of HIV/AIDS in Denmark efficient]. PMID- 9214077 TI - [HIV/AIDS development in Denmark and Sweden]. PMID- 9214078 TI - [Assessment of medical technological possibilities of routine US scanning of pregnant women]. PMID- 9214079 TI - [Intracranial pressure monitoring in patients with severe craniocerebral injury]. AB - After severe head injury intracranial pressure (ICP) must be measured continuously for management to assess and maintain the cerebral perfusion. Therefore in our hospital epidural transducers are used. To prove the efficiency of this method in a 12-month period the clinical courses of 23 patients with intracranial pressure transducers were analysed retrospectively. Eighteen patients survived, 5 of them without residuals, 13 with residuals and 2 remained in coma. In 14 patients secondary rises of intracranial pressure were observed between days 3 and 6 post injury. The mean ICP value of the survivors revealed 25 mm Hg. whereas the expired showed 60 mm Hg. In 17 patients the measurements were considered as reliable, 6 measurements were not reliable, which included 1 of the 5 patients who died. One transduce was displaced, another one showed a hemorrhage at the drill hole. There was no infection. PMID- 9214080 TI - [Rotator cuff rupture. Vascular supply and collagen fiber processes as pathogenetic factors]. AB - Light and polarization microscopic appraisal of the pathways of fibers and blood vessels in the region of the rotator cuff shows branches of the suprascapular artery. These initially radiate into the insertion tendon parallel to the muscle fibers. They do not continue there, i.e. the vessel branches have blind endings, or they branch and anastomose with each other. Outliers of the transverse branch of the anterior circumflex artery of the humerus come from lateral (from the direction of the deltoid muscle). They pass from the bony insertion of the supraspinatus tendon into the tendon plate, but only run together with the fibers for a short distance. Consequently, a zone low in vessels or free of vessels can be constantly demonstrated under a magnifying glass in the course of the supraspinatus and to a small extent also of the infraspinatus in the fetus or neonate as well as in the adult in the region of the zone of interweaving of the tendinous muscle outlier with the capsule at length magnification. In the genesis of rotator cuff rupture, the presence of hypovascularity must be considered to be a predisposing factor which is present from birth onwards. It affects the clinical course during the process of aging as the point of least resistance in consequence of arteriosclerosis, collagen degeneration physiological wear and tear friction at the lower surface of the acromion and inflammatory swellings of the subacromial bursa. PMID- 9214081 TI - [Hydroxyapatite ceramics in clinical application. Histological findings in 23 patients]. AB - Based on the histological findings of 23 patients who had received implants of the bovine hydroxyapatite ceramic Endobon for a period of up to 16 months, the biocompatibility, nature and extent of osseointegration as well as the resorption and degradation behaviour of the ceramic were investigated. The investigation material consisted mainly of small fragments that had been retrieved during revision operations that were indicated for other reasons. The results confirm the good tolerability and suitability that have been systematically investigated in experimental studies and described for hydroxyapatite ceramic as bone substitute in a vital cancellous bone bed that is not exposed to excessive strain (due to its brittle character). The importance of fulfilling certain requirements in order to achieve a successful result, such as stable implantation in a well vascularized, infection-free bone bed also with a minimization of the contact with local connective tissue has been further substantiated. Good success has been achieved by simultaneous loading with autogenous bone marrow as is practised by many ceramic users. In some cases a widening of intergrain boundaries as well as partial dissociation of superficial hydroxyapatite crystallites were observed in the implant surface. PMID- 9214082 TI - [A new technique of segmental bone replacement in metastatic osteolysis in long bones]. AB - Intercalary replacement is a valuable option in the treatment of diaphysary long bone metastases. In certain cases methylmethacrylate bone cement may be used as a cheap and readily available alternative to modular or custom-made prostheses. Stabilization is best performed by locked intramedullary nailing. By cutting open the cylindric part of a large syringe and placing it around the bone ends after resection a mould is created where liquid bone cement can be filled in. Thus spilling of liquid methacrylate into the adjacent soft tissues is prevented. After curing this results in a smooth round intercalary segment with perfect contact to the bone ends allowing early postoperative mobilization. PMID- 9214083 TI - [Apophyseolysis and avulsion fractures of the anterior inferior iliac spine. A case report]. AB - This is a report of a rare case of an avulsion of the apophysis of anterior inferior iliac spine by a 14-year-old football player. Origin, diagnosis, differential diagnosis, and treatment are evaluated and described. PMID- 9214084 TI - [A method for estimating the prevalence of mammary Staphylococcus aureus and Streptococcus agalactiae infections in herds based on an examination of bulk milk samples]. AB - Parallel quantitative determination executed in 92 herds at 107 sampling dates was focused on the counts of major pathogens (Staphylococcus aureus, Streptococcus agalactiae) in bulk milk samples and on the prevalence of the above pathogens in a herd by examination of individual milk samples. The counts of main pathogens were also determined in terms of quantity in rinsing water before milking and in bulk samples in 5 herds with pipeline milking. Tab. I shows the qualitative analysis of the relation. Sensitivity of the method is satisfactory for the pathogens observed (95% and 91%, resp.), but method is less specific for Staph. aureus (67% against 92% Str. agalactiae). Figs. 1 and 2 show coordinate graphs of the results obtained while Figs. 3 and 4 document the distribution of frequency of the particular values in data sets. The values do not exhibit normal distribution (P < 0.01). Spearman's coefficient of rank correlation of bulk milk and individual examinations amounted to 0.823 and 0.900 for Staph. aureus and Str. agalactiae, respectively. Four mathematical models were tested in the course of quantitative analysis, describing the relation between bulk milk examination and individual examination: (1) linear regression, (2) linear regression with fixed starting point, (3) logarithmic regression and (4) irrational function. A model based on the equation of irrational function (4) was found to be best: y = a + bx + c square root of x + k +/- a1 + i(t)c1 square root of x + k1. Tab. II shows the parameters of the equation for examined microorganisms. Correlation coefficients for the above equation are r = 0.733 and r = 0.842 for Staph. aureus and Str. agalactiae, respectively. Prediction curves (Figs. 5 to 8) and confidence regions of prediction curves were also determined for the best model, and a prediction table was constructed (Tab. III). It was confirmed that the milking machines were not a significant source of direct contamination of bulk milk samples with the examined pathogens (Tab. V). PMID- 9214085 TI - [Use of molecular genetics for identification and differentiation of various strains and species of bacteria]. AB - The aim of the paper is to inform on methods and practical application of DNA fingerprinting in typing of living organisms. Methods discussed in the review include standard DNA fingerprinting, plasmid profile analysis, RAPD PCR and direct sequencing of selected parts of genomes. In each method molecular basis together with its advantages and limitations is explained. All described methods are supplemented with selected cases of their practical application. PMID- 9214086 TI - [The effect of a low-protein diet on the epithelium of nephron segments in sheep]. AB - Changes in the epithelium thickness of proximal and distal convoluted tubule of cortex, thin descending limb of Henle loop from inner stripe of outer medulla, thick ascending limb from outer stripe of outer medulla and of collecting duct from inner medulla were investigated in young growing sheep fed a low protein diet (LP). Sheep on LP-diet were given a daily ration with 5.73 g of nitrogen and 6.13 MJ of digestible energy (DE) while the daily intake of control group was 19.51 g of N and 12.29 MJ of DE. Both groups of animals were fed these diets at least 6 weeks before collection of kidneys. Morphometric analysis with digitizing tablet showed that intake of LP-diet resulted in significant reduction of epithelium thickness in both distal convoluted tubule and thick ascending limb while the epithelium of collecting duct was found to be thicker. No changes of the epithelium dimensions were determined in proximal tubule and in thin descending limb of Henle loop. Presented results point to the morphological expression of the adaptation of sheep kidneys to a low dietary protein intake which is associated with the increased renal reabsorption of urea. PMID- 9214087 TI - [Lesions of the peripheral nerves associated with pseudoaneurysms of the major arteries caused by gunshot wounds]. AB - Ten wounded persons, operated on for gunshot-induced peripheral nerve lesion in which pseudoaneurysm of the main artery was found intraoperatively, were presented. Preoperative clinical course, intraoperative neurovascular topography and pathoanatomy, operative procedures and postoperative results were analyzed. The correlation was established between preoperative neurologic deficit and intraoperative findings on arteries and nerves. It was concluded that such patients have to be operated on as soon as possible, but not later than five days from the beginning of neurologic aggravation, to prevent the development of irreversible neural damage. PMID- 9214088 TI - [Reconstruction of defects of the oral cavity with the myocutaneous sternocleidomastoid flap]. AB - The sternocleidomastoid (SCM) muscle can be used for a muscular myocutaneous or osteomyocutaneous flap in the reconstruction of cancer and trauma related defects of the oral cavity. The SCM muscle has 3 principal arterial pedicles and a number of minor ones. The superior pedicle is a branch given off in about 70% of cases by the occipital artery, the middle is formed by a branch of the superior thyroid artery, and the inferior pedicle arises from the suprascapular or the transverse cervical artery. We used the SCM myocutaneous flap for the reconstruction of various defects in the middle and lower part of the face and oral cavity. Group of 28 patients was submitted to transfemoral angiography of the external carotid artery and thyreocervical trunc. The dominant vascularisation of the superior pedicle was found in 17 (60%) cases. We used the SCM myocutaneous flap only in cases with dominant vascularisation from superior pedicle and good results were obtained in all cases. PMID- 9214089 TI - [Ultrasound guided percutaneous biopsy of the kidney]. AB - In the course of 5 years, 582 ultrasound guided percutaneous renal biopsies were performed in 558 patients. Kidney tissue was obtained in 507 patients (90.9%), and in 485 (86.9%) the obtained sample was sufficient to establish the diagnosis. Complications following renal biopsies were observed in 221 patients, or 38% of total biopsies. There were 212 (36.4%) clinically moderate complications. The most frequent ones were asymptomatic hematomae (32.6%), and infrequently lumbar pain (2.4%) and hematuriae lasting less than 12 hours (1.4%). In 9 patients 10 (1.7%) serious clinical complications in the form of hematuria lasting more than 12 h (1%), large perirenal hematomae (0.5%) and urinary infections (0.2%). In the older age group and in patients with pronounced renal failure no significant difference in the incidence of complications was observed. Ultrasound guided percutaneous renal biopsy is a safe diagnostic method, and the associated complications do not seriously curb its use. The therapy of complications is primarily conservative, and only rarely surgical. PMID- 9214091 TI - [The effect of 4-aminopyridine on indirectly evoked contractions of the diaphragm in rats in situ]. AB - The aim of the present study was to investigate the influence of three doses of 4 aminopyridine (1, 3 and 10 mg/kg i.v.) on the amplitude of contraction of rat diaphragm indirectly stimulated by electrical impulses of various frequencies (0.1, 1, 10, 50 and 100 Hz). All the doses used significantly augmented the post tetanic single twitch potentiation after stimulation with 100 Hz (by 35-40%). Doses of 3 and 10 mg/kg i.v. of 4-aminopyridine significantly increased the contractile response to the 10 Hz stimulation (by 73-77%), while only 3 mg/kg i.v. significantly (by 35-39%) increased the amplitude of initial single twitches. It is concluded that the 4-aminopyridine-induced blockade of presynaptic potassium channels caused potentiation of the indirectly evoked contractions of rat diaphragm in situ, which is both dose- and frequency dependent. PMID- 9214090 TI - [Formulation of calcium carbonate tablets with various binding substances]. AB - The test results of calcium carbonate tablets, made of different binding substances (microcrystal cellulose, gelatin, 7pp sodium carboxymethylcellulose and starch) were presented. The content of calcium-carbonate in tablets as well as varying, solidity, prodigality and aptness to decay was determined. The best properties were observed in tablets made with starch. PMID- 9214092 TI - [Disorders of lipid levels and metabolism as risk factors for the onset and development of cerebral arteriosclerosis and cerebral ischemia]. PMID- 9214093 TI - [Use of extracorporeal hybrid (natural-artificial) liver in the treatment of patients with fulminant hepatitis]. PMID- 9214094 TI - [Treatment of basocellular carcinoma]. PMID- 9214095 TI - [Complex partial epilepsy--a differential diagnosis problem]. PMID- 9214096 TI - [Malignant acinar-endocrine cell tumor of the pancreas]. PMID- 9214097 TI - [History of the development of the Academy of Military Medicine]. PMID- 9214098 TI - [Academician Gojko Nikolis--a prominent and important personage in Yugoslav military medicine (I)]. PMID- 9214099 TI - [Author of the year]. PMID- 9214100 TI - [Changes in F-response parameters in supraspinal lesions of the central nervous system]. AB - Since the F response is the result of antidromic activation of limited number of alpha motor neurons, the motor neurons excitability level can be investigated and established by analyzing parameter values of F response. The excitability of alpha motor neurons is the result of balance between excitatory and inhibitory influences of mainly supraspinal origin. The aim of this study was to perform the analysis of basic parameters of the F response in the lesions of pyramidal, extrapyramidal and cerebellar system, and to determine the eventual predictive value of the F response in lesion outcome. The investigations were carried out in 65 patients and 30 neurologically healthy subjects in control group. The patient group consisted of 38 patients with pyramidal lesions due to ischemic stroke, 15 patients with Parkinson's disease and 12 patients with vascular cerebellar lesions. The F response investigations were repeated monthly during 12 months in 12 patients with ischemic stroke. The results showed significant increase in F amplitude and frequency. These changes were in positive correlation with the increase of spastic muscular tone. In patients with cerebellar lesions decreased amplitude and frequency of F response were in positive correlation with the reduction of muscle tone. It was concluded that the measurement of the F response parameters might be reliable marker of the alpha motor neurons excitability and might have predictive value in the early phase of ischemic stroke. PMID- 9214101 TI - [Effect of various types of thermotherapy in the rehabilitation of persons with war injuries of the extremities]. AB - The aim of this study was to check thermotherapy influence on the outcome of the rehabilitation program in patients who suffered contractures of joints following the war injuries of extremities and to compare the three different thermotherapy effects: peloid, hidrocolator and paraffin. The research included 36 examinees divided into 4 parallel groups: three experimental and a control one. The observation features were: movement range, skin temperature and hyperemic halo diameter around the joint. Rehabilitation program lasted ten therapeutic days. Experimental group examinees were treated by kinesiotherapy and various types of thermotherapy. Control group examinees were treated only by kinesiotherapy. The results demonstrated that thermotherapy did not influence significantly the functional recovery. Characteristically the best thermotherapeutical effect to the skin was reached by the use of paraffin. PMID- 9214103 TI - [Use of biological feedback in persons with complete new dental prostheses during the initial period of functional adaptation]. AB - Adductor muscles muscular tone of new complete denture wearers in the period of initial functional adaptation was investigated as well as the use of visual feedback with EMG method for more efficient use of dentures. Thirteen persons of 63 years average were investigated. Action potential of masseter muscle surface layers and pterigoid muscles anterior layers on both sides were registered. Subjects were instructed how to adjust the mandible from the rest position to the full contact with upper denture teeth and to keep it that way for 5 minutes, 3 times a day. Intensity of measured muscle activity was controlled immediately after denture insertion as well as 7, 14 and 21 days after insertion. The greatest amplitude values of all of four examined muscles were registered after denture insertion. Expected changes due to the use of visual feedback had favorable effect on muscular tone lowering during the following period. Maximal therapeutic tone was reestablished. PMID- 9214102 TI - [Standardization of the scale for evaluation of psychopathologic sequelae of traumatic stress]. AB - Horovitz's scale is of questionnaire type and is used to register psychological consequences of traumatic stress with the method of self-assessment. As it is not widely known nor applied in our conditions, we got down to translate, apply and validate the scale in our examinees. The research aim was to check the practicality, applicability, objectivity discriminatory value and validity of the scale and to adopt it for our needs and our examinees who were divided into two groups: participants of war who were expected to show PTSD (posttraumatic stress disorders) (67 examinees) and non-participants of war who might have stress experiences in life (77 examinees). The result of neuropsychiatric examination, results obtained from the PTSD-interview and few other psychological tests of personal capability and personality characteristics were used as a criterion for checking up the validity of the scale. The research results demonstrated the scale to be very simple, economical and easily applicable in various conditions- especially for research, differential diagnosis and follow up effects of therapy on PTSD. In general, reliability and validity of the scale was very high, since it was in positive correlation with the neuropsychiatric examination results, PTSD interview and neuroticism tests. PMID- 9214104 TI - [The lumbosacral syndrome]. AB - The causes of lower back pain were analyzed in 232 outpatients by using the prospective study. The most frequent causes were: degenerative changes, congenital changes, injuries and their sequelae. Among the extravertebral causes, static are the most frequent ones. Pain most often appears because of the already existing pathological substrate. Increased efforts caused by the specific military service emphasized the painful disturbances that could be eliminated by short-term drug therapy. Early diagnosis had the irreplaceable role in professional orientation. Diagnosis such as lumbosacral syndrome should be avoided. The exact cause should be determined thus making a more precise diagnosis and therefore the therapy more successful. Physical treatment combined with the other treatments is of particular significance. PMID- 9214105 TI - [Fascicular anatomy and vascularization of peripheral nerves]. PMID- 9214106 TI - [Risk factors for the development of malignant tumors of the urinary bladder]. PMID- 9214107 TI - [Perirenal hematoma as a complication caused by extracorporeal lithotripsy]. PMID- 9214108 TI - [Association of acute rheumatic fever and acute glomerulonephritis]. PMID- 9214109 TI - [Chronic Q fever]. PMID- 9214110 TI - [Academician Gojko Nikolis--a prominent and important personage in Yugoslav military medicine (II)]. PMID- 9214111 TI - [Oncoepidemiologic situation in the Kaluga region of the Russian Federation ten years after the Chernobyl accident]. AB - Cancer morbidity and mortality in radionuclide-contaminated areas of Kaluga Region have been investigated. The study was mainly concerned with the extent of radiation pollution influence on the present-day cancer morbidity and mortality rates. The correlations between the latter factors, on the one hand, and relevant population risks, on the other, have been analysed. It was found that the present day neoplasm morbidity rates recorded in Chernobyl-stricken areas are chiefly accounted for by a combination of factors which were there before the disaster. The trends of morbidity and mortality from neoplasms of the respiratory tract in females are still unfavorable. There is still no statistical verification of the radiation effect on cancer morbidity except that of thyroid cancer in women in contaminated areas. The morbidity and lethality rates in these areas basically conform to the general trends. The findings are in conformity with those reported worldwide on radiation-related cancer induction latency and biological effects observed in the inhabitants of contaminated regions. PMID- 9214112 TI - [Changes in the optical characteristics of the skin of patients with basal cell carcinoma]. AB - Skin optical characteristics were investigated in connection with large-size tumors and inflammatory infiltrates in 54 patients suffering from basal cell carcinoma. It was found that large size of tumor involves higher light absorption in the skin, lower light conduction and increased photoluminescence. Also, cutaneous tissues look brighter more often when light passes through them. Inflammation in foci of lesions also involves higher light conduction and photoluminescence in the skin. PMID- 9214113 TI - [Treatment of lung metastasis in patients with soft-tissue sarcoma]. AB - The study group included 104 patients with soft-tissue sarcoma and metastases into the lung, treated at the Institute Clinic in 1960-1992. Surgery for lung metastasis was given to 10 and chemotherapy-to 48 patients; the remaining 48 cases received treatment for symptoms only. Overall 5-year survival was 13.5% (after surgery-30.0%; chemotherapy-19.1%). All the 46 patients treated for symptoms died within two years after detection of lung metastasis. Chemotherapy proved effective in the treatment of multiple metastasis when detected shortly after removal of the first lesion. PMID- 9214114 TI - [Correlation of psychoemotional state and membrane condition in patients with lung cancer in the preoperative period]. AB - The growing asthenia of the neuro-psychic system which frequently occurs in cancer patients, particularly, in the preoperative period might undermine the body's defenses and thus contribute to tumor progression. The psycho-emotional system and immunological status of lung cancer patients were corrected with a combination of measures including individual psychotherapy and tranquilizer treatment. A study of blood levels of PI, PIP, PG-F and PG-E2 as well as the use of a number of methods of psychological diagnosis showed that the psycho emotional and immunological status improves as a result of such preoperative therapy. PMID- 9214115 TI - [Surgical and radiation therapy of esophageal cancer]. PMID- 9214116 TI - [Breast cancer with low levels of blood lipids]. AB - Double-bond lipids levels in blood were assessed in 113 breast cancer patients. The levels of double bonds in plasma lipids and red blood cells were found to be much higher than those in healthy females and cases of mastopathia cystica fibrosa. A correlation between tumor extension and insufficient levels of blood lipids was established. The double-bond levels were higher: (a) in blood lipids of inoperable cases as compared with surgical ones; (b) in cases of stage I-II tumor, as compared with stage III, and (c) in cases of unfavorable clinico morphological factors. Blood lipids levels tended to be relatively higher in patients who survived a shorter period after examination. It is suggested that lipids insufficiency in blood may be used as an indicator of certain interrelationships between tumor and body at different stages of tumor process. PMID- 9214117 TI - [Aneuploidy and proliferative activity in breast cancer (flow-cytometric investigation)]. AB - Flow-cytometric assay of DNA levels and proliferative activity of cell in 43 cancers of the breast was carried out. The cytometric data were evaluated versus age, histological malignancy and regional metastasis incidence. The study pointed to a direct correlation between malignancy and aneuploidy (increased fraction of cells featuring unbalanced levels of DNA as well as enhanced DNA index) involved in invasive ductal carcinoma. Aneuploidy and proliferative activity frequency is relatively higher in cases of regionally disseminated tumors. Aneuploidy tumor incidence is higher in age groups under 50 than in older patients. PMID- 9214118 TI - [Determination of the proliferative activity of breast cancer cells using 5-bromo 2'-deoxyuridine in vivo]. AB - Proliferative activity was studied by immunohistological assay of label index (LI) involving the injection of 5-bromo-2'-deoxyuridine in vivo into 10 benign lesions and 40 breast tumors. LI level was found to be significantly higher in breast cancer patients (10.94%) than in cases of benign lesions (1.97%) (p = 0.000002). A similar relation was recorded between relapsed breast cancer (16.65%) and primary tumor (10.28%) (p = 0.03). The study also established a distinct correlation between LI of invasive ductal carcinoma and histological malignancy as determined according to Bloom and Richardson, p = 0.045 between stages I (6.03%) and II (9.60%) and p = 0.01 between stages II and III (15.37%), the ability to form tubular structures (p = 0.003), the numbers of mitoses and hyperchromatic nuclei (p = 0.006) between stages I (5.95%) and II (9.53%) and p = 0.003 between stages II and III (15.00%) and presence or absence of nuclear polymorphism (p = 0.03%). No correlation was observed between LI in breast tumor, on the one hand, and tumor size, degree of invasion, clinical stage, age, menstrual status and estrogen and progesterone receptor levels, on the other. LI in metastasis (7.07%) (15 cases) appeared to be significantly higher than in primary tumor (10.13%) (p = 0.001) and it revealed a distinct correlation with that in primary tumor (p = 0.39). PMID- 9214119 TI - [Clinical evaluation of radiation complications with various methods of radiotherapy for vaginal cancer]. AB - The investigation was concerned with complications development in 133 cases of primary vaginal carcinoma (VC) who had been treated with combined (distance and contact treatment) or low-dosage (LD), medium-dosage (MD) or high-dosage (HD) contact irradiation. Time-dose-fractionation (TDF), cumulative radiation effect (CRE) and linear-quadratic (LQ) ("extrapolated dose of response") were used to evaluate normal tissue response. Early- and late-onset radiation injuries of vaginal mucosa, urinary bladder and rectum were compared vis-a-vis dosage which was calculated on the basis of HDF model. Dosage exceeding the limits of connective tissue tolerance was responsible for the development of early-onset epithelitis, cystitis and rectitis in cases of LD- or MD-brachytherapy of vaginal carcinoma or when the latter was conducted in conjunction with distance therapy. However, early-onset epithelitis and cystitis might appear when dosage remained within tolerance limits. This occurred as a result of HD-contact treatment or when the latter was used as a component of combined therapy. Most late-onset moist vaginal epithelites, rectites and recto-vaginal fistuals were caused by dosage in excess of tolerance. Late- and early-onset vaginal epithelitis was observed with tolerable dosage, too, if HD-brachytherapy alone was used or as a component of combined treatment. PMID- 9214121 TI - [The effect of radiation from videoterminal of personal computer on spontaneous and estradiol dipropionate-induced carcinogenesis in mice]. AB - Two series of experiments using female mice SHR were conducted. In the first run, the animals were exposed to radiation from a video display terminal of a personal computer (VDT), for 1 hr, 5 times a week, starting at the age of 3 months. The treatment continued until their natural death, with and without an Ergostar protective screen filter (SF). During the second run, mice were injected either 10 mkg estradiol-dipropionate (ED), subcutaneously, with 0.1 ml olive oil, or the same dose of olive oil. Beginning from the first injection of either substance, the mice started to receive VDT radiation like those of the first series did. The frequency of persistent estrus appeared to be significantly higher in the first group, irrespective of whether SF was used. The latter factor had virtually no bearing on tumor incidence rate and tumor latency period in this experiment. However, experiment 2 established a noticeable increase in incidence of mammary and ovarian tumors matched by a shorter latency period in ED- and VDT-treated animals as compared with intact controls. At the same time, mice exposed to ED and VDT+SF revealed fewer tumors of the reproductive system. It was suggested that there is no relationship between exposure to VDT and spontaneous tumorigenesis, irrespective of whether SF is used or not. However, exposure to VDT may slightly promote development of ED-induced tumors, while application of SF may moderate the process. PMID- 9214120 TI - [Hemopoietic stem cell transplantation in lymphogranulomatosis]. AB - Twenty patients with resistant and relapsed Hodgkin's disease were included into this investigation. The treatment effectiveness and regimen-related toxicity of BEAM-protocol were evaluated. Stem cell transplantation (SCT) was carried out either as bone marrow transplantation (BMT) in 11 patients or peripheral blood stem cell transplantation (PBSCT) in 9 patients. A comparison of the results pointed to a significant decrease in neutropenia duration, fewer blood transfusions needed and shorter stay in hospital in the PBSCT group. Complete remission was observed in 90% of patients of the SCT group, 91%-in BMT and 89% PBSCT groups. It was found that high-dose chemotherapy followed by SCT is more effective in the treatment of resistant and relapsed Hodgkin's disease and, in certain situations, it is a treatment of choice. As to the toxicity of BEAM protocol, it does not exceed those of the other chemotherapy modalities used in SCT. PMID- 9214122 TI - [Sonography in the diagnosis of lesions of abdominal lymph nodes, liver and spleen in malignant lymphoma]. AB - The data on clinico-sonographic examination of 603 patients with Hodgkin's disease and non Hodgkin is disease are presented. Involvement of abdominal lymph nodes was recorded in 186 (30.8%) cases. The diagnostic procedure specificity and sensitivity in Hodgkin's and non-Hodgkin's disease were 98.5; 64.5 and 99.6; 86.1%, respectively. The results of sonography application in detecting malignant lymphomas in the liver and spleen proved uninspiring because only foci could be identified which accounted for not more than 6-10% of all lesions of different organs. Diffuse lesions were detected in a few cases only. PMID- 9214123 TI - [The results of cytologic diagnosis of thyroid diseases (20 years experience)]. AB - The data on fine-needle aspiration biopsies and imprint smears for thyroid nodules and regional lymph nodes carried out in 1129 patients at the Institute Clinic (1976-1995) were reviewed. Histological examination was performed in 340 cases. The cytological procedure showed a diagnostic sensitivity of 92.3% and a specificity of 75.8%. PMID- 9214124 TI - [Dimorphous cancer of the lung]. AB - Dimorphous cancer of the lung with predominantly peripheral localization was diagnosed in 9 (1.2%) out of 716 patients operated for lung tumor in 1986-1995. The blood-serum levels of neuron-specific enolase, tissue polypeptide antigen, carcinoembryonic antigen, alpha-fetoptotein, CA 19-9 and CA-125 carbohydrate antigens and total activity of alkaline phosphatase were assayed in all the patients before surgery. The study showed an 1.5-times increase in CEA concentration in 5 cases, with primary tumor size being 5 cm and more. PMID- 9214125 TI - [The quality of medical care given to patients with esophageal cancer in St. Petersburg and the Leningrad region]. AB - The data on the morbidity, time of diagnosis and results of treatment of esophageal cancer in St. Petersburg and Leningrad Region (1992-1994) were analyzed. The morbidity rates were higher in women than in men, the predominant age being over 60. Morphological verification data were not available in almost half the cases. Stage I-II tumors were detected in 15.7-26.5%. Specialized treatment was given to 25%, combination one-to nearly 2%. To raise the standards of medical aid, it is suggested that measures be taken to ensure stable operation of the Cancer Register, St. Petersburg, and to offer treatment to esophageal cancer patients at specialized medical facilities. PMID- 9214127 TI - [The purposefulness of splenectomy as "a matter of principle" during surgery for stomach cancer]. AB - The findings on radical surgery for gastric tumors T3 and T4 performed in 315 cases were analyzed to evaluate the effects of splenectomy. No influence of splenectomy on short- and long term results was observed. No justification for splenectomy as an operation of principle was found. PMID- 9214126 TI - [Factors determining treatment effectiveness in localized stomach cancer]. AB - The report deals with the data on the results of radical combination surgery in 315 patients with stomach tumors T3 and T4. Extensive and combination surgery coupled with a well-balanced surgical, anasthesiological and medicinal therapy proved the only effective method of managing localized cancer of the stomach. PMID- 9214128 TI - [The potential for rehabilitation of patients with proximal gastric cancer]. PMID- 9214130 TI - [Experience with low-field magnetic resonance tomography for the diagnosis of bone marrow lesions]. AB - Metastatic lesions of the bone marrow (BM) in cancer patients are instrumental in making prognosis and selecting optimal treatment modality. Recent evidence has pointed to insufficient sensitivity of morphological, X-ray and osteoscintigraphic diagnostic procedures to deal with focal alterations in BM. Vistas in application of low-field magnetic resonance tomography for detection and differential diagnosis of localized lesions in BM are discussed. PMID- 9214129 TI - [Method of primary jejunogastroplasty after gastrectomy and subtotal resection of the distal stomach for cancer]. AB - Post-gastrectomy and post-resection conditions are occurring more frequently due to the growing number of radical operations carried out for stomach cancer. The available procedures for restoring the natural continuity of the gastrointestinal tract are either too sophisticated or ineffective. A new technique of primary jejunogastroplasty following subtotal distal resection of the stomach is suggested. Application of newly-devised esophagoen-terostomy and enteroduodenostomy makes possible storage and passage of chyme into the duodenum and prevents refluxesophagitis and reflux-jejunitis from developing. A 3 month-3 year follow-up of 35 surgical patients showed that a number of grave post gastrectomy complications could be avoided if the procedure had been used. PMID- 9214131 TI - [Ultrasonic diagnosis of nodal lesions of the axillary-jugular lymphatic plexus and their evolution]. AB - Axillary lymph nodes of three levels may be adequately differentiated on the basis of the TNM classification by application of layer-by-layer tomographic examination of axillary fossa and thoracic and clavicular-thoracic triangles in the axillary cavity. Echographs show musculus thoracis minor as a distinct point of reference. Topographic features of deep lateral cervical lymph nodes may be used in anatomical identification of internal jugular nodes, transverse cervical artery, accessory nerve and suprajugular lymph nodes. In cases of multiple involvement of lymph nodes, their shape (chain or cluster) may be identified. It can be shown Whether lymph flows are in series, collateral centripetal, collateral centrifugal (recirculatory) or run parallel in metastatic areas. Varying patterns of lymph flow through collateral pathways formed in the axillary jugular lymph plexus are responsible for the variety of patterns of lesion clusters in the lymph nodes system. PMID- 9214132 TI - [Lasting disease-free course of synovial sarcoma]. PMID- 9214133 TI - [Successful surgical treatment of cancer with multiple location]. AB - The lecture deals with the specific characteristics of cell membrane damage caused by in vivo and in vitro irradiation at the dosage commonly used in radiation therapy of malignant tumors. The mechanisms of lymphoid cell loss to ionizing radiation are described. They include metabolic depletion, direct damage to the cell membrane causing loss of its asymmetry and surface receptors expression being altered. PMID- 9214134 TI - [Cellular mechanisms of functional disorders of the lymphatic system in high-dose radiotherapy]. PMID- 9214135 TI - [Effectiveness of peroral application of a new immunocorrective agent of natural origin]. AB - Data on peroral application of new peptide origin immunocorrector from nervous tissue of hydrobionts are cited. It has been established that immunological gangliin activity, perorally introduced, is close to that being applied parenterally. This drug comprises a soft immunocorrector that is expressed by humoral and cellular factor strengthening of immunity, increasing induces of phagocynosis of PMNC and sells SMPh. Peroral gangliin introduction protects 50 60% of mice from fatal coli-infection. Gangliin availability to be taken perorally opens the door to its wide application as food addition. PMID- 9214136 TI - [A rapid method of energy metabolism assessment in the human body using thermovision technology]. AB - Quick method of energy metabolism assessment by direct calorimetry using thermovision technology for momentary registration of thermoreturn from whole human body surface is suggested. For realization this method an algorithm of computer program was developed for processing hundred thousands of points of skin temperature measures and temperature of surrounding environment. This method can be used for assessment of big groups of population. The method was protected by patent. PMID- 9214137 TI - [Effect of soybean products on the reproductive system of sexually mature female albino rats]. AB - Adult albino rats were fed with standard ration with 15% soybean products addition during four months. The "soy-milk", after the short-term boiling, and "soy-milk-UHF", after the ultra-high frequency influence, were investigated. The reproductive system of female rats were studied. It was found that soybean products included into the diet changed the sexual cycle, increased the postimplantation embryonic mortality, decreased the posterity body weight during the first 14 days of life. The results of the investigations conducted have shown that ultra-high frequency influence on soybean product reduces it's reproductive system effects. PMID- 9214138 TI - [Effectiveness of using processed cheese enriched with iodine in the prevention of endemic goiter]. PMID- 9214140 TI - [Change in postprandial glycemia as affected by various carbohydrate-containing products in patients with type II diabetes mellitus]. AB - The postprandial glycaemic effect and glycaemic indexes of some dishes from cereals and grains were studied in patients with type II diabetes. The studied food had different glycaemic indexes. The need of testing traditional Russian food and dishes for patients with II type diabetes is discussed. PMID- 9214139 TI - [Experience in using the biologically active additive "miprovit" in the treatment of various metabolic diseases]. AB - Using of protein and vitamin enriched preparation "Miprovit" in treatment of 90 patients with obesity showed decreasing effect on serum cholesterol and glucose levels and coefficient of atherogenity which were increased in these patients. Decreasing of appetite and feeling of hungry were noted in 75% of patients. "Miprovit" favoured more comfortable feeling of patients during course of body weight reduction. PMID- 9214142 TI - [Evaluation of the quality of food products and rations]. AB - The article delivers information about method of calculation from estimation quality food products and rations nourishment from different contingents feeding. Investigations conduct on the example meat-fish wares. Elaborated and scientific grounded permit biology of full valve products nourishment. PMID- 9214141 TI - [Monitoring food consumption and nutritional status of Moscow schoolchildren in 1992-1994. 2. Anthropometric evaluation of nutritional status, effect of social factors on the character and status of nutrition]. AB - In this part of the study the nutritional status of Moscow's schoolchildren was assessed by height and weight. The anthropometric data were compared with the CDC/WHO international growth references standards by ANTHRO version 1.01 software. The prevalence of low weight-for-age (Z-score < -2) was more frequent in boys of 15 years of age. Low height-for-age was more prevalent in the group of boys of 15 and girls of 10 years of age. In the period under study there was a slight decrease in height and weight of schoolchildren, but this was within the limits of expected normal variation. A few selected socio-economic variables such as parental education, family size, participation in organized sports and use of school breakfast/lunch options were included. The size of the family or whether it is a one- or two-parent household does not seem to influence energy or nutrient intake, nor are the anthropometric variables significantly affected. The more size of family and the lower the father's education than lower the children's total fat intake and intake of energy from fat. PMID- 9214143 TI - [Caffeine and children]. AB - Beverages containing caffeine are consumed by most people in most countries most days. Consumption is mostly in beverages such as coffee, tea and some soft drinks, and smaller amounts from other foods such as chocolate. Children also consume caffeine, though in smaller amounts even relative to their smaller size. Many questions have been asked about possible health effects of caffeine and have been answered by scientific research. Studies on pregnant women consuming caffeine show no effects on the fetus, infants, or on development followed up to school age. There have been many studies on children of school age. For example, it has been shown that a single dose of 3 mg/kg is without appreciable effect on a variety of behavioral and physiological functions, and even 10 mg/kg, had only minimal effects, within the normal range of differences between the children without caffeine. While newborn infants metabolize caffeine slowly, children from less than 1 year to adolescence metabolize caffeine about twice as fast as non smoking adults. The numerous studies showing safety of caffeine in adults, combined with the direct studies in children showing they are similar and not more susceptible to caffeine than adults, gives assurance that lifelong consumption of caffeine in foods and beverages, starting in childhood, is without deleterious effects on health. PMID- 9214144 TI - [Socio-economic status of the family. Effect on nutrition and health of the child]. PMID- 9214145 TI - [The sugar manufacturing complex in Russia]. PMID- 9214146 TI - [Advance directives from the medical viewpoint]. AB - Advance directives, enabling patients to declare their wishes and preferences for future situations, when they no longer are able to communicate, appear to be attractive. The concept, however, is based on two important presuppositions: First on the recognition of the right of self-determination. Second, it is assumed that a patient can direct medical care not only hic et nunc, but can advance autonomy into future incompetent states. It is the second presupposition that causes most of the ethical concerns and practical problems. Some of the limitations can possibly be overcome, or at least minimized, by careful drafting, frequent revision and the appointment of a proxy. However, advance directives should be integrated into the process of patient information and care. PMID- 9214147 TI - [Acceptance and validation of advanced directives--ethical and clinical considerations]. AB - Crucial decisions of end-of-life treatment quite often are handed over to machines and other technical capabilities of prolonging life at any cost. Various forms of advance directives have been developed, but not widely accepted by the lay public and rarely recognized by physicians and the health care establishment. We discuss the benefits and risks of different forms of advance directives and present, based on own experience and on evaluating the clinical-ethical and clinical-legal debates, a model combining information on value-and-wish status, the designation of a surrogate decision maker, and a limited number of recommendations or directives for crucial end-of-life decisions. We call for further research on physician's and patient's attitudes towards end-of-life decisions and the development and validation of advance directives, as we feel that it will be the culture of physician-patient interaction rather than legal measures that will change ethical and cultural attitudes and medical procedures. PMID- 9214148 TI - [Values anamnesis: a narrative approach to determining and interpreting advance directives]. AB - Quite a number of conceptual, clinical, ethical and legal arguments have been made to agree in favour of advance directives for medical treatment and care. But not many patients execute advance directives and physicians and nurses are quite reluctant to accept advance medical directives as authoritative guidance for clinical decision making in cases of dementia, terminal illness or in the case of the dying. PMID- 9214149 TI - [Theological moral considerations of advance directives]. AB - The forms for advanced directives vary considerably in respect to content and wording. This article intends to develop ethical criteria for such texts, from the moraltheological point of view, also considering the social context. PMID- 9214150 TI - [Advance directives in Austria law]. AB - This article discusses legal aspects of advance directives. In Austrian law there is only one provision: section 10 Hospitals Act ("medical record"). Therefore permissibility, contents and form of advance directives are of special, interest. Concerning the liability of advance directives the author concludes that this question has to be answered case by case. PMID- 9214152 TI - At risk drivers: are physicians required to report? PMID- 9214151 TI - [General practice of advance directives in the USA]. AB - DEFINITION OF THE PROBLEM: Advance directives are written documents which tell what a person wants or does not want if he/she in the future cannot make his/her wishes known about medical treatment. There are three major forms of advance directives in the US: living will, durable power of attorney, and medical directive. Even though advance directives have been used for over 25 years, probably no more than 20% of Americans have prepared a written directive. What are the reasons for this relatively small percentage? It has been argued that they run the opposing risks of either being too general or too specific and that they often use a vague and confusing terminology. Furthermore, the two most frequently cited barriers were the patient's expectation that the physician should take the initiative and the sense that such issues were only relevant for those who were older or in worse health. Progress in developing useful advance directives cannot stop with existing documents. A critical evaluation of advance directives and the development of new ideas will be necessary. Therein, the most important area for future efforts is empirical research. PMID- 9214153 TI - Delayed arrival of George Orwell's 1984. PMID- 9214154 TI - Time to update Uniform Controlled Substances Act. PMID- 9214155 TI - Current Perspectives in Acid Inhibitory Therapy. New Haven, Connecticut, November 17-19, 1995. Proceedings of a conference. PMID- 9214156 TI - [Training program for diabetic feet]. PMID- 9214157 TI - [Patellar chondromalacia as a pathomorphological finding. There are no positive clinical or radiological signs]. PMID- 9214158 TI - [Sports following endoprosthetic joint replacement]. PMID- 9214159 TI - [Psychological factors in the therapy of lumbar disk herniation]. PMID- 9214161 TI - [Movement training in the prevention of postural weakness]. PMID- 9214160 TI - [Are there magnetic resonance tomographic changes following shock-wave treatment of tendinitis calcarea?]. PMID- 9214162 TI - [Stress complaints in musicians]. PMID- 9214163 TI - [Cruciate reconstruction using an 8-fold plantaris tendon]. PMID- 9214164 TI - [Hip endoprosthesis in young patients]. PMID- 9214165 TI - [Early implantation of a knee endoprosthesis in young rheumatic patients?]. PMID- 9214166 TI - [Current role of ultrasonography in orthopedics. Results of a nationwide survey]. AB - The purpose of this study was to determine the current position of diagnostic ultrasound imaging in the day to day orthopaedic practice. A questionnaire was sent to all the 387 conservative and operative orthopaedic hospitals and orthopaedic university departments in Germany. 167 of these hospitals completed the forms and provided the basis for this evaluation (response rate 43.2%). Screening of the fetal hip for CDH was by far the most common indication, the shoulder was ranking second, the knee and the hip joint in children and adults rank third and fourth. Other frequent indications were injuries of the achilles tendon, the menisci of the knee, the knee in rheumatoid arthritis and muscle injuries. Ultrasound imaging is clearly established as a diagnostic tool at least in the named areas. It is therefore of importance to include ultrasound examination techniques in the curriculum of the orthopaedic training scheme. PMID- 9214167 TI - [Correlation of pathological clinical hip symptoms in the sonographic study of infant hips]. AB - The relevance of clinical signs of infant hip dysplasia and the importance of a sonographic newborn-hip screening are often discussed. Between 1984 and 1990 at our department 6532 infant hips were examined both clinically and sonographically. Sonographic hip findings were pathological in 71.7% of the hips with positive clinical Roser-Ortolani sign and in 100% of the hips with positive Hilgenreiner sign. In contrast, pathological hip types according to Graf were seen only in 18% of the hips with limitations of the abductions of the hip joint. Examining other clinical signs for infant hip dysplasia within this study, as well we can conclude, that only the Roser-Ortolani sign and that of Hilgenreiner showed a high specificity for infant hip dysplasia, while most of the clinical signs e.g. limitation of abduction in the hip joint are rather unspecific. PMID- 9214168 TI - [Sonographic diagnosis of anterior syndesmosis rupture]. AB - In a prospective trial we evaluated the diagnostic value of ultrasound imaging in the diagnosis of injuries to the anterior syndesmosis of the ankle joint. In an initial series of 20 patients using a basic standard ultrasound machine (Siemens Sonoline SL 1 with a 7.5 MHz piece) and a simple dynamic testing protocol as described in this article we found a significant difference between the ultrasound result for the ruptured syndesmosis versus non rupture and a specificity similar to that of arthrography. We conclude that ultrasound imaging is a useful tool in the diagnosis of syndesmotic rupture. Given the statistical significance of these results we feel confident that-despite the small amount of patients of this series and after an individual learning curve-ultrasound will eventually replace arthrography as the standard of investigation for this type of injury. PMID- 9214169 TI - [Three-dimensional computer reconstruction of the pelvic and hip area in the preoperative planning of orthopedic interventions and surgical simulation]. AB - RESEARCH QUESTION: Osteotomies are elective operations which require exact pre operative planing. Computer assisted three-dimensional planning of orthopedic procedures requires a three-dimensional reconstruction of the region of interest. To be applicable, such a three-dimensional reconstruction must be an accurate mathematical description of the region of interest, and not only a 3-D-image. The purpose of this study was to create such an accurate 3-D reconstruction. METHODS: We describe a method to create an exact reconstruction from CT-data, from the data acquisition, contour determination by an algorithm, triangulation of the resulting data and reconstruction of the region of interest. CONCLUSION: The described method is a useful tool to obtain exact individual three-dimensional reconstructions of any skeletal region of interest. This makes pre-operative planning and simulation of the results of orthopedic procedures at the computer possible. PMID- 9214170 TI - [Comparison of roentgenological, macroscopic and histological degree of degeneration in varus gonarthrosis]. AB - In 20 cases of varus gonarthrosis the degree of severity of osteoarthrosis was assessed preoperatively, intraoperatively and postoperatively for the medial and lateral compartments, respectively. The preoperative assessment was carried out on the basis of x-ray taken in an anterior-posterior and lateral view, whereas the intraoperative assessment was based on the recognizable macroscopic alteration of the femur and tibia. Following implantation of a total knee prosthesis, the resected osteocartilogical samples were assessed from a histological point of view. In the case of varus gonarthrosis there is a distinct difference in the degree of degeneration in the medial and lateral compartments. This difference becomes less noticeable if the assessment is carried out intraoperatively. The histological analysis shows that there are certainly differences between the compartments from a quantitative point of view but a late stage of osteoarthrosis was discernible both medially and laterally. From a histological point of view the radiological concept of the so-called "unicompartmental varus gonarthrosis" cannot be supported. In both compartments the osteoarthrotic process is more uniform than is to be expected. PMID- 9214171 TI - [Meniscus replacement using incongruent transplants--an experimental study]. AB - The effect of incongruous medial meniscal grafts on the articular surface was studied. We compared the cartilage degeneration in 4 Groups of 25 sheep knees. The histological changes were graded according to the score of Mankin, the morphological alterations by the score of Jackson. In Group 1 no surgery was performed. In group 2, the meniscus was totally detached from its base at the capsule and refixed without changes in the congruity or isometry. This group provided the basic data. In group 3, the contralateral medial meniscus was used as a transplant by reversing it. The reattachment was done according to isometric conditions. With this technique only the congruity of the tibial and femoral surface was modified. In group 4, the medial meniscus was completely removed. The results of group 1 (Mankin grade 0.58; Jackson grade 0.00) and group 2 (grade 0.50 and 0.47) showed no difference. The highest degree of degenerative changes occurred in the meniscectomized knees (group 4, grade 4.47 and 1.73), however considerable changes were also found in the knees with an incongruous meniscal graft (group 3, grade 3.37 and 1.27). The results suggest that incongruous grafts will lead to degeneration of the articular surface but still have a chondroprotective effect. PMID- 9214172 TI - [The principle of autogeneic rib perichondrial transplantation in the treatment of deep articular cartilage defects]. AB - The purpose of this study was to examine the fate of autologous perichondrial grafts after transplantation into cartilage lesions in weight-bearing joints. Results were evaluated depending on the age of the animals and on the weight bearing conditions. Osteochondral lesions were drilled in the articular surface of knee joints in 36 adolescent sheep. The defects were filled with autologous rib perichondrial grafts which were secured by either collagen sponges (n = 12) or fibrin glue (n = 12). Twelve animals served as controls. Following one week of immobilisation the animals were allowed to move freely. Animals were sacrificed after 4, 8, 12, and 16 weeks. The same procedure was performed in three adult animals in order to achieve hints regarding the age-dependent ingrowth of the transplants. In two of them the transplantation was done using fibrin glue for fixation; one animal served as control. Grafts were removed corresponding to the time intervals and investigated histologically. In adolescent animals grafts from weight-bearing areas and control defects did not show a regular cartilagenous differentiation. In contrast to that, hyaline-like cartilage formation could be noted in non-weight-bearing areas even after 4 weeks. In principle, the same results were found in adult sheep but a delay of cartilagenous differentiation of four weeks was observed. Depending on these results we use the procedure of perichondrial transplantation clinically for treatment of circumscript deep lesions of articular cartilage provided no osteoarthritis or any other overlying general diseases are evident. By means of the Lysholm- and Ranawat-score all six patients demonstrated postoperative improvement. Furthermore, in two patients arthroscopic controls exhibited sufficient filling of the defects and ingrowth to the surrounding cartilage. Thus, autogeneic perichondrial transplantation can be recommended for treatment of circumscript deep cartilagenous lesions of articular cartilage. PMID- 9214173 TI - [High-energy extracorporeal shock-wave therapy (ESWT) in the treatment of pseudarthrosis]. AB - PROBLEM: The success rate of high-energy extracorporal shock wave therapy in the treatment of non-unions in comparison to the "golden standard" surgery is still unclear. METHOD: In a prospective study, 3000 impulses with an energy density of 0.6 mJ/mm2 were applied with an experimental device to the pseudarthrosis in 52 patients. RESULTS: The mean duration of pseudarthrosis was 13 months. A mean of 2.5 surgical interventions had already been performed. Bony union was achieved in 52% of our patients after an average of 3.3 months. Failures especially were found in the atrophic type of pseudarthrosis as well as in congenital bone disorders like fibrous dysplasia or osteogenesis imperfecta. No serious complications were observed. CONCLUSIONS: Even after numerous surgical interventions high-energy extracorporal shock wave therapy as a noninvasive method for the treatment of bony non-unions showed a fair success rate. A higher success rate can be expected by strict selection criteria. PMID- 9214174 TI - [Correction of high-grade sesamoid bone dislocation in hallux valgus using Austin's osteotomy with and without lateral soft tissue release]. AB - INTRODUCTION: Aim of this study was to analyze if, using the Austin technique for correction of hallux valgus deformity, the additional soft tissue procedure is capable to achieve better correction of the sesamoid subluxation, the hallux valgus angle and the intermetatarsal angle. PATIENTS AND METHODS: 19 patients with 20 feet operated according to the original Austin technique and 26 patients with 28 feet operated according to a modified technique with lateral soft tissue release both with a preoperative sesamoid subluxation grade 3 were compared with the help of a standardized questionnaire in respect to clinical and radiological results. RESULTS: Analyzing the clinical outcome of the two procedures, there was no statistical difference. Comparison of the radiological results revealed a significantly better correction of the sesamoid position and a better correction of hallux valgus and intermetatarsal angle by using the additional soft tissue procedure. DISCUSSION: The lateral soft tissue procedure with release of the adductor hallucis, dissection of the deep transverse plantar ligament and mobilisation of the sesamoids is capable of a significantly better correction of the sesamoid position. Only a combination of osseus and soft tissue correction is capable to correct the pathomechanism and to guarantee long lasting results. PMID- 9214175 TI - [Is the orthopedically correct lifting technique rational from a cardiopulmonary and metabolic viewpoint?]. AB - INTRODUCTION: In spite of educational work for many years by the orthopaedic surgeons wrong lifting techniques are still used in everyday life as well as during exercise. The reason for this could be the fact that there is an advantage regarding these techniques from the energetic point of view. MATERIAL AND METHODS: To clarify this issue we examined 30 healthy males within the age range of 17 and 30 years (weight: 74.0 +/- 9.2 kg, height: 182.5 +/- 6.7 cm). In a randomised sequence the volunteers carried out two different lifting techniques: 1. Lifting with stretched legs and bent back, 2. Lifting with bent legs and stretched back (Brugger technique). The test scheme consisted of four different 3 minute exercise levels with increasing weights (0 kg, 5.2 kg, 12.1 kg, 19.2 kg), the repetition rate was 30 times per level. RESULTS: Using the Brugger technique the results for heart rate, VO2, VCO2 and VE were highly significant (p < 0.001) above the results of the other technique at all exercise levels. On the RPE (rate of perceived exertion) scale the differences were statistically significant regarding the 0.01-level. CONCLUSION: The results give rise to the supposition that the energy consumption for the lifting technique with the bent back is lower and is therefore preferred in everyday life and during exercise. These findings should be taken into consideration when informing people about possible damages. PMID- 9214176 TI - [The effect of halo-gravity traction in the preoperative treatment of neuromuscular scoliosis]. AB - The purpose of this study is to evaluate the influence of halo-gravity-traction on paralytic scoliosis in various neurologic diseases. Between 1980 and 1993 preoperative halo-gravity-traction was applied in 32 patients with paralytic scoliosis (23 patients with myelomeningocele, 6 patients with poliomyelitis, 3 patients with cerebral palsy). In the myelomeningocele group the average curvature before treatment was 97.8 degrees, after surgery 45.1 degrees; which is an improvement of 53.9%. Halo-gravity-traction accounted for 12.8% improvement. In the poliomyelitis group the average curvature before treatment was 104.3 degrees, after surgery 58.0 degrees; which is an improvement of 44.4%. Halo gravity-traction accounted for 16.9% improvement. In the cerebral palsy group the average curvature before treatment was 75.0 degrees, after surgery 39.0 degrees; which is an improvement of 48.0%. Halo-gravity-traction accounted for -2.7% improvement. If there is an effect the question remains, whether this will have consequences for the surgical outcome. The comparison between good and bad responders with the surgical result shows, that this result is independent of the halo-gravity-traction. From this results we draw the conclusion, that preoperative halo-gravity-traction can not be recommended in paralytic scoliosis. PMID- 9214177 TI - [Lymphoma-induced imitation of knee prosthesis loosening]. AB - Persistent pain after knee arthroplasty is often caused by aseptic loosening, implant failure, unphysiological alignment (especially of the patella), infection, scars and neuroma. In a case of a 70-year-old patient pain and swelling after knee arthroplasty persisted despite three revision procedures. Surprisingly, a non-Hodgkin's lymphoma was found in femoral bone marrow histology. A metastatic non-Hodgkin's lymphoma as cause of pain and swelling after knee arthroplasty, like in this case, is rare and has not been described up to now. PMID- 9214179 TI - [Quality assurance]. PMID- 9214178 TI - [Proteus syndrome: a case report]. AB - The Proteus-syndrome is a recently described congenital hamartomatosis consisting of numerous clinical features of great variety. Mainly affected are the musculo skeletal system, primarily by hemihypertrophy, macrodactyly and exostoses, and the skin and the subcutaneous tissue, primarily by pigmented naevi and subcutaneous tumors. The differential diagnosis includes other malformation syndromes, e. g. Klippel-Trenaunay-Weber syndrome and other hamartomatosis. Surgical intervention and treatment is difficult because of a frequency of complications and recurrences. This article describes clinical manifestations of Proteus syndrome, differential diagnosis and therapeutic strategies. PMID- 9214180 TI - [Characteristics in the treatment of scoliosis in muscular diseases]. AB - INTRODUCTION: Patients suffering from the most frequent muscle disorders Duchenne muscular dystrophy (DMD) and spinal muscular atrophies (SMA), who ceased walking respectively are confined from the outset to the wheel-chair, are developing commonly a progressive scoliosis (collapsing spine) due to an increasing muscle weakness. Basing on the pelvic obliquity these scolioses are leading first of all to problems in sitting as well as difficulties in trunk and head control. Along with the increasing weakness of respiratory muscles these phenomena entail a restrictive respiratory insufficiency. CONSERVATIVE TREATMENT: An effective conservative treatment is not available for these scolioses. The use of a corset, however, can only be taken into consideration as a compromise, either for very young patients or those who refused an operation respectively who have reached an inoperable stage. The exclusive use of so-called "anatomic sitting supports" in the wheel-chair in order to treat or prevent a progressive scoliosis in DMD or SMA is absolutely to be rejected. They should only be applied for very young patients with SMA type II as a transitional solution until a corset or better an surgical stabilisation of the spine will be effected, or as a palliative measure in late stages. SURGICAL TREATMENT: Only the early as possible performed surgical stabilisation of the spine using adequate instrumentation (Luque, CD or modifications), enabling an early mobilization without corset or cast, is the most effective treatment of these scoliosis. Patients with DMD or SMA type III should be stabilized after loss of walking ability and definitive confinement to wheel-chair, if the curve is more than 20 degrees-30 degrees Cobb and progressive and forced vital capacity (FVC) is > 35%. The instrumentation should be applied between D3 or D4 and sacrum. The bony fusion mass should include the lumbar and lumbosacral region. The unfused instrumentation with the telescope-rod after Naumann is a good solution for patients with SMA type II and progressive curves already in the early childhood from ca. 5 years of age. First of all surgical spinal stabilisation improves the sitting comfort. Over and above this the improved cosmetic appearance should not be underestimated for the psychological condition of these patients. Additionally it is proved, that surgical stabilisation of the spine prolongs the life expectancy of patients with DMD. Furthermore stabilization of lung function can be achieved for both DMD and SMA patients in comparison to the natural history of these diseases. Altogether a decisive improvement of quality of life can be reached for all these patients. PMID- 9214181 TI - [The characteristics of the structure of night sleep in patients who have had a stroke]. AB - The aim of the study was investigation of the structure of the night sleep in relation to the disease stage and location of the destructive focus. 18 patients with ischemic stroke (in the right hemisphere-8 cases, in the left one-6 persons, with brain stem location-4 patients) were observed as well as 5 practically healthy individuals. The diagnosis was verified by computer tomography in all cases. Clinical neurological investigation and polygraphy of night sleep (electroencephalography, electrooculography, electromyography) were performed. Analysis of sleep parameters was carried out according to program elaborated by the Center of Sleep Research of Sechenov Moscow Medical Academy. This program includes the analysis of segmental sleep structure in addition to standard parameters. The sleep quality was estimated by Sleep Index (Slind) according to the degree of intactness or destruction of its structure. Rude disorganization of night sleep structure was revealed in the patients, including disturbances of mechanisms of the whole sleep organisation, as well as generation and maintenance of its separate stages. The most rude disorders of sleep in patients with ischemic stroke were connected with location of ischemic focus in right hemisphere and in medial, deep structures. PMID- 9214182 TI - [Reflex sympathetic dystrophy]. AB - 35 patients with syndrome of reflex sympathetic dystrophy (RSD) were examined clinically by means of psychological tests, method of evoked skin sympathetic potentials (ESSP) and nociceptive flexor reflex R III. RSD syndrome had central origin (after strokes) in 17 patients (group I) while in 18 patients it arose after some peripheral damages (group II). RSD syndrome was characterised in both groups by triad of symptoms: acute pain, autonomic disorders and osteoporosis. Decreased life quality, high anxiety and depression were quite characteristic for the patients with RSD. Both significant prolongation of latent period and decrease of amplitudes of ESSP were observed on the damaged limb in both groups of patients. These damages were evidently connected with disorders of central peripheral autonomic interactions in paretic limbs in group I, in group II they were conditioned by damages in peripheral sympathetic sudomotor fibers. Significant increase of both thresholds of pain perception and flexor reflex was found in patients with RSD while the value of the coefficient of their correlation was decreased. Such disorder might be caused by initial insufficiency of antinociceptive functions and that permitted to suggest the hypothesis about the role of this factor in pathogenesis of RSD syndrome and in development of chronic pain its leading symptom. The conclusion was drawn that development of RSD syndrome was not associated with location, severity and character of the damage, but it was conditioned mainly by the state of functions of cerebral antinociceptive systems as well as by emotional and personal peculiarities of the patients. PMID- 9214183 TI - [The adaptive capacities of patients with panic disorders (a psychophysiological study)]. AB - Contingent-negative deviation (CND) and orientational reaction (OR) were studied in 9 women suffering from typical (according to DSM-III-R) panic disorders (PD) and 12 healthy females. CND recording was made after giving neutral (70 dB, 1000 Hz) and stress (120 dB, 2000 Hz) pairs of sounds. Patients with PD were divided in two groups depending upon CND amplitudes. The first group (4 patients) was distinguished from healthy individuals by high CND amplitude (which significantly decreased after reaction on stress sound), by delay of extinction of cutaneogalvanic reaction (CGR) and by higher electromyogram amplitude. The second group (5 patients with more severe course of PD) was characterized by low amplitudes of CND and electromyogram, by absence of changes in CND after stress sound, by delay of extinction of CGR and nonspecific response. Above-mentioned data were evidence of psychophysiological heterogeneity of typical PD. The results were interpreted from a position of adaptive abilities of patients with PD, using ineffective and immature psychophysiological strategies of overcoming stress. PMID- 9214184 TI - [The psychopathology of a depressive somato-autonomic symptom complex]. AB - The investigation is based on the hypothesis about psychopathological heterogeneity of the somatovegetative (biological) symptom complex of depression. To testify this statement the results of the analysis of the somatovegetative component of depressive syndrome in randomized selection (20 adult patients) were presented. Somatovegetative symptoms (disorders of sleep and appetite mainly) defined the pattern of debut and preceded manifestations of other signs of affective disorder in this group. Psychopathological qualification of disorders belonging to such symptom complex was presented as the phenomena of alienation of the most deep vital psychic functions, exactly somato-vegetative drives. The main psychopathological statements were argued by the data about dynamics of such states, which were followed by transformation into typical vital depressions. The results obtained were interpreted in accordance with traditional K. Schneider's conception of "depression without depression". PMID- 9214185 TI - [The psychopathology of paroxysm-like disorders in endogenous mental diseases]. AB - 45 patients with endogenous mental disorders were observed: 11 with manic depressive psychosis and 34 with shift-like progredient schizophrenia. The peculiarity of the cases observed was the presence of paroxysmal-like disorders (PD) in clinical picture of the disease. 8 variants of PD were established: 1) with vegetative disorders, 2) with affective disturbances, 3) with obsessions, 4) with sensopathies and depersonalization, 5) with impulsive drives, 6) with alterations of consciousness (similar hysteric twilight type), 7) with hallucinations, 8) with development of manifestations of Kandinskii-Clerambault's syndrome. The duration of PD varied from several seldom till a few hours, while their frequency from 1-2 in a year till several in twenty four hours. Paroxysmal like states were accompanied by anxiety, fear or by low- differentiated sense of discomfort and "suppression", but second subjective affective feeling was neutral or even pleasant. The phenomenon which might be called as the anxious waiting for the fit was also marked. The observations of combinations of different variations of PD in one person as well as the cases of including of elements one variant in the picture of the other one permitted one to suggest the presence of a single pathobiological basis for PD rise. PMID- 9214186 TI - [The psychosomatic correlations in bronchial asthma]. AB - Combination of somatic symptoms with neurotic manifestations and personality's disturbances in clinical picture of bronchial asthma was established during clinical and psychological observation of 89 patients with bronchial asthma. It was shown that increase of the number of frustrating situations (when psychical vulnerability was elevated), strengthening of both anxiety and emotional tension, as well as rigidity of negative emotions, hypochondriac and anxious-depressive tendencies composed the whole correlational system with alterations in functions of external respiration, changes in blood immunoglobulins levels just as with clinical indices of bronchial asthma. This system represented different levels of psychosomatic correlations' regulation. The complex psychophysiological factor of frustration and emotional tension was described, moreover the increase of its value was accompanied by strengthening of both psychical alterations and somatic disorders which were quite characteristic for prevalence of either trophotropic activation or ergotropic one when beta-adrenoceptors were blocked. PMID- 9214187 TI - ["Music of the Brain" in the treatment of insomnia patients]. AB - The effects of "Music of the Brain" which is the new nonpharmacological method of treatment of insomnia were studied. The method is based upon the transformation of EEG signals into the music by using a special algorithm developed by the author. Sleep polygramme was registered and analysed. EEG sites of sleep stages and phases which most satisfied usual criteria were selected and transformed into music. The patient listened to the recorded audio cassette just before sleep. 58 patients with insomnia were divided into 2 groups according clinical, formal, psychological and electrophysiological (polysomnography, EEG) methods which were applied before and after 15-day course of treatment. 1st group (44 individuals) was the main group and the 2nd group (14 patients) was the "placebo" group ("Music of the Brain" of other individual was used in this case). The positive effects of "Music of the Brain" were evaluated in patients with insomnia in more than 80% cases. These effects were marked on the basis of both subjective feeling and objective (psychological and neurophysiological) results. The high effectivity of "Music of the Brain" for patients with insomnia combines with the lack of side effects and complications. PMID- 9214188 TI - [The medicine of sleep in the neurological light]. PMID- 9214189 TI - [Instenon in the treatment of acute disorders of the cerebral circulation]. AB - Effectiveness of Instenon (Alkaloid Pharm. Co.), the preparation, containing hexobendin, hydroxytheophyllin, etamivan was investigated in treatment of patients with acute cerebrovascular disorders. Instenon was applied in acute phase of ischemic stroke both in form of ampules and dragee as well as for intravenous injections too. Marked positive influence of Instenon was observed on motor, sensory, autonomic and psychic functions. Instenon in form of ampules had advantage compared to dragee form in terms of faster positive alterations onset. Application of Instenon in form of ampules was more effective in patients with disorders of consciousness while usage of drug in the form of dragee was more effective in patients with cerebrovascular disorders in vertebro-basilar system. The drug hadn't significant influence on systemic arterial pressure either after single injection or long-termed treatment; the mild hypotensive effect was observed after therapeutical session. The drug was well tolerated by patients in both forms and hadn't any side effects. There were no negative consequences of Instenon interaction with other preparations. The conclusion was made about possibility of application of Instenon for the treatment of ischemic stroke. PMID- 9214190 TI - [Caffetin in the treatment of neck pain and lumbago-sciatica]. AB - The effectiveness of caffetin, combined analgetic drug, was estimated in treatment of acute cervicalgia and lumbar ischialgia (1 tablet 3 times daily during 10 days). Both clinical and psychological parameters were analysed. Caffetin had pronounced analgetic effect which was observed on the 2-3d day of drug application. Meanwhile there was improvement of clinical and psychological parameters, namely: relaxation of the muscles, decrease of either tension symptoms or of the levels of anxiety and depression as well as improvement of quality of life. The patients well tolerated caffetin administration without any side effects. There were no negative consequences of interaction of caffetin with other drugs (vascular, psychotropic). The conclusion was drawn about safety and effectiveness of caffetin for alleviation of acute pains in patients with myofascial, myotonic and radicular syndromes of different origin. PMID- 9214191 TI - [The effect of Imovane on sleep structure and respiratory indices during sleep in insomnia patients]. AB - Night sleep was examined in 120 patients with different forms of insomnia. The positive effect of Imovane was shown both on presomnic and intrasomnic disorders. Transitory side effects (dryness, bitter taste in the mouth) were in some cases that didn't need cessation of therapy. Polysomnographic studies confirmed effectivity of Imovane: there were observed either lengthening of the duration of sleep, its 2 stage, delta-sleep, or decrease of the duration of falling asleep. There were no respiratory changes after course of Imovane, meanwhile some indices positive dynamics was revealed in this case. PMID- 9214192 TI - [A comparative clinical evaluation of the antidepressive activity of fluoxetine and fluvoxamine]. AB - Comparative study of the peculiarities of clinical action of fluoxetine and fluvoxamine in 65 patients with endogenous depressions revealed their high efficiency (in 74.3% and 64.3% respectively). Fluoxetine was characterised by predominance of a stimulating effect from the first days of treatment as well as by relatively late manifestation of very thymoleptic and tranquilizing impact (during 3-4 weeks). Fluvoxamin displayed relatively uniform occurrence of separate clinical effects together with predominance and early appearance of antidepressive influence. On the basis of the comparison of the peculiarities of either clinical action of fluoxetine and fluvoxamin or their side effects with those of traditional antidepressive drugs (amitryptilin and ludiomil) the preferable indications for their prescription were determined. Thus fluoxetine was very good in treatment of apathetic-adynamic depressions while fluvoxamin was recommended for therapy of anxious and melancholic depressions. Antidepressants studied were ranked in the following way in terms of decrease of sedative effect and increase of stimulating action: amitryptilin, fluvoxamin, ludiomil, fluoxetine. The proper thymoleptic effect of fluoxetine and fluvoxamin exceeded the same effect of amitryptilin and ludiomil. PMID- 9214193 TI - [Patterns in the change of the evoked skin autonomic potential in nervous system diseases]. AB - The study of sympathetic skin response (SSR) was performed both in 30 healthy volunteers either in rest or after pharmacological preparations administration (reserpine, sydnocarb, gammalone) and in 48 patients with autonomic disfunction, syringomyelia, polyneuropathy. It was determined that two processes were presented in SSR structure, exactly: the first one which included I and III phases and was bound with trophotropic functions and the second process which was presented by II phase only and reflected the activity of ergotropic functions. The latent period of SSR reflected the tonus of the sympathetic nervous system. It was established that SSR changed regularly in the nervous systems diseases studied: thus, increase or decrease of the latent period as well as of the amplitudes of SSR phases were observed in autonomic disfunction while depression or absence of SSR took place in syringomyelia or polyneuropathies. The data obtained testified the results of the other authors about possibility of SSR usage for diagnosis of both central and peripheric autonomic disturbances. PMID- 9214194 TI - [A quantitative assessment of the blood supply to the head by indirect rheography]. AB - Clinical-experimental basis of the possibility of application of the method of indirect rheography of head (IRH) in wide clinical practice was presented. Estimation of the results reproduction and accuracy of the method was performed too. 40 healthy individuals of both sexes at the age of 18-50 and 102 patients with spinal osteochondrosis, arterial hypertension and neurocirculatory dystonia were examined. It was determined that IRH provided 93% of results reproduction and its precision was 90% and higher in comparison with ultrasound method. It was established that head circulation was 750-1200 ml/min or 12-24% of cardiac output in healthy individuals. The results of the study of head blood circulation in some groups of patients were also discussed. PMID- 9214195 TI - [Evoked potentials in parkinsonism]. AB - Evoked potentials (EP) were analysed in patients with idiopathic form of Parkinson's disease (PD) in terms of three modalities: visual EP of chess pattern, acoustic EP of brain stem and somato-sensory EP during stimulation of median and tibial nerves. The data were compared with clinical characteristics. The conclusion was drawn about evident participation of afferent systems in PD development which is apparently secondary. PMID- 9214196 TI - [Focal dystonias]. PMID- 9214197 TI - [The molecular genetic and toxicological-ecological bases of the etiology and pathogenesis of Parkinson's diseases (parkinsonism)]. PMID- 9214198 TI - [The typology of the objective disorders of night sleep in insomnia]. AB - The most frequent polygraphic manifestations of insomnia were analysed in 39 neurotic patients with disorders of night sleep (NS). The frequency of pathological alterations of the objective characteristics of NS was determined after examination of the data obtained. Four types of sleep alterations were recognized. There was not always clear correlation between the subjective estimation of sleep and its objective characteristics. For choice of optimal therapeutic policy it was proposed to evaluate leading complaints of the patients for more accurate definition of the degree of expression of objective manifestations of insomnia without usage of NS polygraphy. Possible mechanisms of NS disorders were determined. Classification of different types of objective disorders of sleep in insomnia was elaborated. PMID- 9214199 TI - [Problems of mental health protection in the work of the 10th World Congress of Psychiatry]. PMID- 9214200 TI - [A scientific and practical conference on affective and obsessive-phobic disorders. Their clinical picture, therapy, neurophysiology and epidemiology]. PMID- 9214201 TI - [A new embedding material for morphometry: Shiojirin E-10]. AB - For morphometric studies, sections have been stained with the Luxol fast blue-PAS Hematoxylin (LPH) staining method after secondary fixation with chromic acid followed by embedding in Celloidin to measure the area of nerve cells or axons with the help of an image analyzer and a microscope. However, the manufacture of Celloidin has been suspended in Japan. Therefore, we studied the use of Shiojirin E-10 (10% nitrocellulose ether alcohol solution) as a substitute embedding material, and compared it with Celloidin from the viewpoint of tissue shrinkage and staining characteristics. Regarding the shrinkage ratio of LPH-stained sections of human cauda equina, there was no practical difference in the axonal areas (p < 0.01) between Celloidin sections (4.31 +/- 2.55 microns2) and Shiojirin E-10 sections (4.14 +/- 2.20 microns2). Regarding staining characteristics, we introduced a hue analysis method using computerized digital signals converted from pictures of stained sections taken by a videocamera. For the evaluation of hue, we examined the H-E sections of mouse liver and kidney, and the LPH sections of the human cauda equina. The hematoxylin hues of the H-E sections were 295 degrees for Celloidin and 294 degrees for Shiojirin E-10. Eosin hues of the E-E sections were 320 degrees for both embedding material. The axonal hues of the LPH stain were 254 degrees for Celloidin and 251 degrees for Shiojirin E-10. Myelin sheath hues were 224 degrees for both embedding material. According to our results, Shiojirin E-10 can be substituted for Celloidin from the qualitative and quantitative viewpoints in the morphologic studies. PMID- 9214202 TI - [Giant cell tumors of the spine. Report of a case, literature review]. AB - Giant cell tumors of bone are uncommon in the vertebrae above the sacrum. We report the case of a giant cell tumor of the third lumbar vertebra, revealed by lumbar and radicular pain. X ray, computed tomography and magnetic resonance imaging showed osteolysis of the body and vertebral arch of L3. Histologic evaluation gave a conclusion of a giant cell tumor, grade 2. Spondylectomy of L3 was performed using a combined approach (anterior and posterior) in two stages. The patient had a good functional result without recurrence at three years and six months. A review of the literature indicates that the radiological appearance is nonspecific but shows the extent of the tumor. Diagnosis can be based only on histological features. Radiotherapy could induce malignant transformation. Radical resection limits the risk of recurrence. Total spondylectomy is recommended for giant cell tumors when both the body and arch are involved. PMID- 9214203 TI - [Malignant hemangiopericytoma of the pelvis: treatment using internal hemipelvectomy]. AB - Hemangiopericytoma is a rare vascular tumor originating from pericytes. The main manifestation is a growing mass. Although computed tomodensitometry and MRI are the best diagnostic procedures, the diagnosis is made only after microscopic examination. The malignancy of this tumor is not well established. In fact, there is a continuum between benign and malignant hemangiopericytoma. The malignancy is estimated by the number of mitoses and the cellularity. Recurrences occur in 60% of cases. The best treatment is surgical excision. The role of radiation therapy and chemotherapy is not well established. The history of a 51-year-old woman treated by internal hemipelvectomy is presented here. PMID- 9214205 TI - [Treatment of benign prostatic hyperplasia with finasteride. Results after 7 years of follow-up]. AB - Our centre took part in an international, multicentre, Phase III trial with Finasteride (MK-906) in the treatment of BPH. In the first year, this was a double-blind, randomised, placebo-controlled study which included 25 patients who were randomly assigned: 9 to treatment with placebo, 8 to finasteride 1 mg and 8 to finasteride 5 mg. After the first year, all patients were assigned to finasteride 5 mg as a single daily dose taken before breakfast. Of the 25 patients who started the trial, 7 have completed 7 years treatment (28%): 1 from the placebo group. 1 from finasteride 1 mg, and 5 who took finasteride 5 mg throughout the study. Total symptoms score, assessed by the modified Boyarsky questionnaire, improved during the first year 4.97 points (51%) and remained unchanged up to year 7 with a final reduction of 6.2 points (64%). Percentual increase in peak urinary flow during follow-up ranged between 21 and 45.9% with an absolute increase at 7 years of 4.2 mL/seg (47.5%) and a reduction in prostate volume of 26%. Finasteride tolerance was excellent at all times, and no serious clinical reaction was seen in laboratory parameters. Two patients reported decreased libido and sexual potency and 1 decreased libido. In summary, since after 7 years of treatment efficacy is maintained in at least 30% of patients with an excellent safety profile, finasteride can be considered an alternative in the medical treatment of BPH. PMID- 9214204 TI - [Impact of prostatic benign hyperplasia and prostatic inflammation on the increase of prostate specific antigen levels]. AB - PSA is currently the best marker in the diagnosis and staging of prostate cancer, although it has a low specificity when trying to distinguish between BPH and prostate cancer. Between January 1994 and December 1995, 243 BPHs were diagnosed after prostate TUR and retropubic adenomectomy. A selection of 131 cases were analyzed based on PSA higher than 4 and normal rectal examination pre-surgery. After surgery PSA was determined again with a time interval from the earlier one ranging between 103-211 days, noting that in about 70% cases PSA levels were normalized and 64% of these also presented focused acute prostatitis, chronic prostatitis, prostate infarction, lithiasis or areas of abscess formation. In spite of a significant number of patients with high PSA levels, this elevation should be interpreted cautiously considering the large percentage of cases where posterior pathological anatomy is a sign of BPH. We believe that among the conditions that could justify such abnormal PSA elevations are those described of areas of chronic or acute prostatitis, prostate infarctions, areas of abscess formation and presence of intraprostate lithiasis. PMID- 9214206 TI - [Quantification of urine leaks: standardized one-hour pad test]. AB - OBJECTIVE: To assess the feasibility and diagnostic performance of the one hour pad test proposed by ICS. PATIENTS AND METHODS: The pad test was performed in 20 women referred for incontinence. Five were included in a surgical protocol and 15 had inconsistent medical history. Their mean age was 52.5 years, the mean time of incontinence was 3.4 years and they used a mean of 3.5 pads/day. Six showed leakage on physical examination (PE) and 2 during previous cystometry. RESULTS: The test is well tolerated and lasts one hour and 15 minutes. The test was positive in 17 (mean leakage = 168 g). Leakage was not related to duration of incontinence, number of pads or cystometric capacity. A weak negative correlation (r = -0.52) was found between leakage and voided volume on uroflowmetry performed after the test. All patients who leaked on PE had a positive test and a significantly higher leakage compared to patients who didn't leak on PE. CONCLUSION: The pad test is a feasible outpatient procedure in the context of a Urodynamics Unit, providing objective information on leakage in most patients. PMID- 9214207 TI - [Analysis of the reliability of ultrasonic estimates of the posturination+ residue]. AB - OBJECTIVES: The most precise procedure for the measurement of residual urine volume is bladder catheterization; nevertheless, it takes real associated morbidity. Our objective is to define the accuracy of bladder ultrasonography when residual volume estimation is performed. METHODS: Thirty patients were studied. Ultrasound estimation of residual volume was undertaken with Simpson's method V = (pi/6) x L x T x AP. Obtained values were contrasted with the post catheterization registered ones. RESULTS: The mean ultrasound estimated residual urine volume was 82.1 ml., versus 156.5 after catheterization. The mean difference was 74.4 ml. (significant, p < 0.001). Correlation coefficient was 0.93. Regression equation was calculated, a well as the corrected formula that, according to the outputs of our study, offers greater reliability in the estimate of the aforementioned volume. Ultrasound sensitivity when proper estimation of volumes equal or greater to 100 cc. is carried out was only 47%, opposite to the elevated specificity (100%) for this purpose. CONCLUSIONS: Transabdominal ultrasound underestimates, to a large extent, the values of residual urine volume. We consider that ultrasound estimation of residual volume constitutes an alternative to be kept in mind; nevertheless, the formula to calculate this volume through bladder diameters introduces an important error and so, it should be corrected. PMID- 9214208 TI - [The use of a continent urinary stoma in complex reconstructions of the lower urinary tract in children]. AB - Since its description in 1980, the Mitrofanoff principle has become a widely utilized and successful technique for the management of patients with a variety of urological disorders. We report our experience with this procedure in 14 patients (10 M, 4 F). The age range was 3.5 years to 17 years (average 12 y) and follow-up was from 6 months to 3 years (average 1.7 y). Patients were classified in 2 groups: I) When this procedure was done because of the patient was unable to perform urethral catheterization (8p). II) Concomitant bladder neck transection and Mitrofanoff diversion (6p). The appendice was used in 9p, ileum in 1 and ureter in 4. Bladder augmentation was performed with ureter in 2p and colon sigmoid in 4. In 1p, ileo-cecal segment and in other colon+ileum, were used to replace the bladder. All patients catheterize the Mitrofanoff channel easily, there were no case of stomal stenosis and the conduit was continent in all. We consider that Mitrofanoff principle is a very successful technique and it can be used as the primary continence mechanism or as an adjunct of major urinary tract reconstruction, to ensure complete bladder emptying, in patients unable to perform urethral catheterization. PMID- 9214209 TI - [Prevention of prolonged drug-induced erection by intracavernous injection of etilefrine]. AB - Here in we report the results of intracavernous injection of etilefrine hydrochloride in a group of 12 patients who developed rigid erection after a diagnostic test with intracavernous PGE1 injection or cavernosometry. Etilefrine reversed erection in all cases. No side-effects or significant changes in blood pressure were noted. Etilefrine is believed to be a safe and low cost alternative to other alpha-adrenergic agents in the prevention of drug-induced priapism. PMID- 9214210 TI - [Surgical solutions to iatrogenic ureteral lesions in 31 cases]. AB - OBJECTIVE: To conduct a critical review of the etiological, clinical and diagnostic aspects of iatrogenic ureteral lesions caused by various surgical disciplines, primarily focused on the different therapeutical options used. PATIENTS AND METHOD: Between January 1985 and December 1995, 31 iatrogenic lesions of the ureter, 19 female and 10 male, mean age 52 years, were examined. The right ureter was involved in 19 cases and the left one in 12 (2 bilateral). The ureteral lesion resulted from gynaecological surgery in 19 cases (61%), general surgery in 6 (19%), vascular surgery in 3 (10%) and urology in 3 (10%). In 9 cases (29%) it was diagnosed during surgery: in 14 cases (45%) as a result of clinical signs and symptoms and in 8 cases (25%) casually. Surgery was the option chosen in 22 cases, while in 9 conservative treatment was followed. RESULTS: Surgery solved 91% of damaged ureters, and the best results were seen in patients where lesion was repaired immediately. Conservative treatment was resolutive in all cases. CONCLUSIONS: Gynaecological surgery continues to be the most frequent cause of iatrogenic lesions of the ureter. Ureteral damage in males is now increasing due to the demands from general oncologic surgery, which is usually appreciated by the surgeon which allows immediate repair. Lesions caused by gynaecologic and vascular surgery are commonly overlooked and are diagnosed post-surgically either because of their clinical manifestations or accidentally. The different surgical techniques currently available allow a successful repair of this kind of lesions. Conservative therapy is appropriate in cases that fulfil a series of basic requirements. PMID- 9214211 TI - [Genitourinary malacoplakia]. AB - Malacoplakia is a chronic inflammatory disease the etiology of which remains obscure. It has a very low incidence and affects primarily the genitourinary tract, although it has been described in some other organs. This paper presents a historic insight of the clinical cases diagnosed in this centre, and includes a review and update of several issues related to this entity such as pathogenesis, pathological anatomy and treatment. Also, the peculiarities related to the involvement of each separate organ with regard to diagnosis, prognosis and treatment are described. PMID- 9214212 TI - [Meconial hydrocele]. AB - Presentation of one case of meconial hydrocele, a very infrequent disease that should be taken into account in all newborns presenting intrascrotal mass. Ultrasonography performed to a 29 year-old female during the 36th week of pregnancy, demonstrated in the fetus the presence of an enlarged right hemiscrotum with homogenous material inside, which was diagnosed as an intrascrotal haematoma due to a likely intrauterine spermatic cord torsion. After eutocic delivery, within one month from birth the newborn developed signs and symptoms which were compatible with acute scrotum and the emergency surgical exploration showed meconial hydrocele secondary to patency of peritoneal-vaginal duct with associated inguinal hernia. The causes, forms of presentation, differential diagnoses and therapeutical options of meconial peritonitis, a rare condition with favourable evolution in most cases, are analyzed showing that, under certain circumstances, treatment is controversial. PMID- 9214213 TI - [Emphysematous pyelonephritis]. AB - OBJECTIVE: To present an uncommon but startling case of a urinary tract infectious disease such as renal emphysematous pyelonephritis. METHOD/RESULTS: We report the case of a decompensated diabetic female patient who presented an intrarenal infectious picture with gas formation compatible with emphysematous pyelonephritis that showed good evolution following medical treatment and simple nephrectomy surgery. CONCLUSIONS: Emphysematous pyelonephritis is a serious infective disease that occurs almost exclusively in diabetic patients and requires a fast and correct diagnosis differentiating between diffuse from focused processes in order to advise conservative or aggressive therapy on the renal unit (percutaneous drainage/nephrectomy), based on immediate hemodynamic and antibiotic medical support, due to its serious prognosis such as is described in the literature. PMID- 9214214 TI - [Wunderlich's syndrome: report of 3 cases]. AB - Report of three cases of Wunderlich Syndrome secondary to renal adenocarcinoma, hydronephrosis and non-filar blood dyscrasia, highlighting its uncommon etiology. Brief review of the etiopathogenic, diagnostic and therapeutical aspects, pointing out the relevance of renal ultrasonography and, most specially, abdominal CAT to achieve a definitive diagnosis and to determine the attitude to follow, trying to use conservative surgery whenever possible, although our three cases were treated with nephrectomy. PMID- 9214215 TI - [Epidermoid cysts of the testis]. AB - The epidermoid cyst of the penis is an uncommon benign intratesticular tumour. This paper presents two cases that were diagnosed and operated in our centre. It also describes the most significant clinical, radiological and histological aspects. PMID- 9214216 TI - [Alkaptonuria, prostatic calculi, and ectopic ureter]. AB - Alkaptonuria is an uncommon tyrosine metabolism disorder. Deficit of homogentisic acid oxidase leads to the elimination of large amounts of homogentisic acid in the urine with accumulation of homogentisic acid oxidized pigment in the connective tissue (ochronosis). The ultimate evolution of ochronosis is degenerative arthritis. The clinical case reported is a 57-year old male diagnosed with alkaptonuria in an early stage of the connective tissue disease who comes to the clinic due to a lower urinary obstructive syndrome secondary to benign prostate hyperplasia but is diagnosed with giant prostate lithiasis. The patient undergoes retropubic adenomectomy with lithiasis removal and re implantation of ectopic ureter from a dual left excretory system. Clinical evolution is completely successful. The rarity of the case calls for circulation and revision of the clinical and therapeutical aspects of this entity. PMID- 9214217 TI - [Molluscum contagiosum in immunodepressed patient]. AB - Molluscum contagiosum is a skin lesion caused by a poxviridae virus. Infection in children takes place through cutaneous contact while in adults results from sexual contact or under immunosuppression conditions. In immunosuppressed patients this infection is caused by opportunistic microorganisms and, basically, its appears when CD4 levels reach a certain decline, therefore its presence can be considered as a marker for poor prognosis or advanced stage of the disease. PMID- 9214218 TI - [National Commission on Urology]. PMID- 9214219 TI - [In vitro study with techniques of imaging of the composition of urinary calculi]. AB - Pre-treatment knowledge of the lithiasic composition can be useful to design the most appropriate therapeutic scheme for each kind of stone. The relationship between the stone's densitometry information provided by the different imaging techniques, conventional radiology (RX), computerized axial tomography (CAT) and dual energy radiographic densitometry (DO) is analyzed, as well as the elemental composition determined by the microanalysis of fragments obtained post-lithotrity using a scanning electronic microscope (SEM) associated to X-ray dispersion energy (XDE). 60 stones, 12 for each pure composition selected (calcium oxalate mono and dihydro, phosphocarbonate, magnesium ammonium phosphate and uric acid), were studied with XR, CAT and DO and were later subjected to lithofragmentation in vitro. Fragments analysis was carried out post-lithotrity with SEM associated to XDE. The X-ray does not allow to establish the composition of some calculi. CAT quantifies the mineral contents of the oxalocalcic and infective calculi and differentiates the uric acid from the other compositions because the mean density values are under 500 Hounsfield Units. DO evaluates the lithiasic content in phosphocarbonate salts which are structurally similar to bone hydroxyapatite. PMID- 9214220 TI - [Pain in children. An unsettled problem]. PMID- 9214221 TI - [Use of DNA for the identification of newborns]. PMID- 9214222 TI - [Neisseria meningitidis strains with decreased susceptibility to penicillin and ampicillin]. AB - Thirty-eight patients (31 children and 7 adults) with meningococcal infection (sepsis and/or meningitis) were studied. The strain most frequently isolated was B (44.7%), followed by C (31.6%). Of the strains isolated, 52.6% were moderately resistant to penicillin (91.6% if only strain C was considered). No resistance to cephotaxime or chloramphenicol was found. Even though patients with moderately resistant strains treated with penicillin G evolved satisfactorily (minimum inhibitory concentrations 0.12-0.50 microgram/ml), the possible appearance of more resistant strains and/or of strains that produce beta-lactamase leads us to the conclusion that cephotaxime is the treatment of choice until an antibiogram is available. PMID- 9214223 TI - [Cystic dilation of the bile duct in childhood]. AB - OBJECTIVES: Common bile duct dilatation (CBDD) represents part of a wide spectrum of pancreaticobiliary disorders, with different etiopathogenic mechanisms. The objective of this study was to compile the cases treated in our service during the last five years. PATIENTS AND METHODS: Four cases of CBDD (17 months to 10 years of age) are reported. All of them presented abdominal pain and bilious vomiting. One patient previously had pancreatitis. Cholestatic jaundice was associated in only one patient. The diagnosis was made by ultrasound, being confirmed by endoscopic retrograde cholangiopancreatography (ERCP) in three cases and by computed tomography scan (CT) in one case. RESULTS: Three patients had a single fusiform dilation of the extrahepatic bile duct (type I cyst, Alonso Lej Todani classification), which were treated by cyst excision and hepaticojejunostomy by using a Roux-en-Y limb. In one patient, the ERCP detected a combined dilatation of the intra- and extrahepatic bile duct (type IV cyst), associated with an anomalous choledochopancreticoductal junction with a distal obstruction of the common bile duct. In the case, the treatment consisted of a transduodenal esfintherotomy. CONCLUSIONS: Based on our experience and a literature review, an increasing incidence of this pathology is deduce. Therefore, the relevance of ultrasounds and ERCP in the diagnosis and visualization of pancreatobiliary ducts and the choice of treatment, depending on the CBDD, are discussed. PMID- 9214224 TI - [The evaluation of fat and muscles of the arm in the nutritional study in pre school children in Madrid]. AB - OBJECTIVES: The objective of this study was to evaluate the body composition of the pre-school children in the city of Madrid as a part of their nutritional evaluation. MATERIAL AND METHODS: Seven hundred and three healthy children of both sexes were studied by means of a cross-sectional sample. The children, between 2 and 6 years of age, attended nursery schools in the city of Madrid. The study evaluated the arm muscle and fat areas through an anthropometric assessment. After measuring the triceps skinfold and the arm circumference, we obtained the arm muscle circumference, the arm area, and the arm muscle and fat areas. Once all data were gathered, they were statistically evaluated. RESULTS: The parameters studied reveal a normal distribution in the population in question. The mean values of muscle and fat areas were normal in relationship to age. PMID- 9214225 TI - [Epidemiological study of the symptomatology of childhood depression in Madrid school-age population]. AB - OBJECTIVES: This study shows the results of an incidental analysis of the prevalence rate of childhood depression (dysthymia and major depression) obtained in an epidemiological cross-study. MATERIAL AND METHODS: This study was carried out among Madrid's school-aged population from primary schools (3 degrees, 4 degrees and 5 degrees EGB). The sample included a total of 1,275 children, both boys and girls, between 8 and 11 years old. They were chosen at random from different school systems (public, semi-public and private) and educational levels (3 degrees, 4 degrees and 5 degrees EGB). The definition of "case" is based on Poznanski's semi-structured clinical interview (Children's Depression Rating Scale-Revised, 1984) depending on DSM-III-R's operational criterion for dysthymia and major depression symptomatology. RESULTS: The prevalence rates obtained in the general school-aged population were 6.1% for dysthymia and 4% with major depression. The total rate of depressive disorders was 10.1%. PMID- 9214226 TI - [Prospective study of 288 cases of acute appendicitis during childhood: characteristics in preschool children]. AB - OBJECTIVE: The purpose of this study was to verify that preschool children with acute appendicitis show some clinical characteristics which, associated with the faster evolution of the infection at this age, result in a higher incidence of perforations, peritonitis and complications. PATIENTS AND METHODS: A group of 288 children between 9 months and 17 years of age with acute appendicitis was studied over a 17-month period, ending July 1993. They were divided into two age groups: Group I (< 5 years: n = 45) and Group II (5 or more years: n = 243). Comparisons between clinical, laboratory and radiological findings, appendiceal pathology, microbiology and complications were made. RESULTS: Children fro Group I showed a higher incidence of perforations (29% vs 7.8%), peritonitis (69% vs 36%), appendiceal masses (37% vs 10.2%), positive cultures (66% vs 18%) and complications (24% vs 9.8%) than those from Group II. All differences were found to be significant (p < 0.05). Children from Group I more frequently showed a set of clinical characteristics forming an atypical picture consisting of: 1) Diffuse abdominal pain (69% vs 30%); 2) Associated infections with non-specific symptoms (33% vs 11.5%); 3) Previous therapy with antimicrobial agents (40% vs 9.8%); 4) X ray findings compatible with gastroenteritis (27% vs 7.4%); and 5) Inability of the child to specify the intensity and location of pain. CONCLUSIONS: All these factors justified the delay in the diagnosis and its significant relationship with the higher incidence of peritonitis in this age group. PMID- 9214227 TI - [Turner's syndrome: growth, endocrinological study and other aspects]. AB - OBJECTIVE: This was work was designed to analyze, in a group of patients with Turner's syndrome treated at our hospital, the following issues: 1. To follow the evolution of several common values of body growth from birth to adult life, as well as the influence of growth hormone therapy. 2. To determine the changes in gonadotrophins and alterations of the thyroid gland. 3. To define the relationship between the karyotype and the development of menarche. MATERIALS AND METHODS: Neonatal growth values were obtained from 32 patients. Body size before and after the treatment with growth hormone was determined in 20 cases. We determined the values of TSH/T41 and FSH/LH in 35 and 34 cases, respectively. In 34 patients older than 15 years and not treated with estrogens, we evaluated the relationship between the karyotype and the presence of menarche/amenorrhea. RESULTS: Mean values of body length, weight and cranial perimeter were 477 mm, 2,877 g and 335 mm, respectively. These values were below those of the normal population, especially for those patients with a 45.X karyotype. The mean height for adult patients was 141.6 cm (range 133.3- 152 cm). After 2 years of treatment with growth hormone the growth velocity changed from -0.25 to +3.55 SD. We did not find pathological values for TSH or T4 (3.3 mU/l and 1.85 ng%, respectively). The LH and FSH values were higher in 64% and 88% of the patients, respectively. These differences were higher in patients older than 13 years, which showed values of 20 and 60 U/l. Twenty-three percent of the patients had the menarche and none were 45.X. CONCLUSION: The early detection of Turner's syndrome permits the physician to follow these patients, improving their growth and their endocrinological control. PMID- 9214228 TI - [Influence of diet in idiopathic hypercalciuria]. AB - OBJECTIVES: To find out if differences in diet may justify the presence of idiopathic hypercalciuria (IH) and to verify if the calcium-restricted diet, often recommended for treatment of IH, is nutritionally appropriate. PATIENTS AND METHODS: Dietary intake and mineral and electrolyte urinary excretion were studied twice in 10 children diagnosed of IH and in 9 controls. First, while being on a free diet. Second, after a week of suppression of milk and dairy products. RESULTS: Intakes of calories, proteins, carbohydrates, fats, and minerals were similar in IH patients and controls. Withdrawal of dairy products of diet resulted in an important reduction in the intakes of calcium (71.1%), phosphorus (63.5%) and fats (30.1%), although only calcium intake was remarkably below international recommendations (33.6 +/- 10.4%). CONCLUSIONS: Spontaneous diet of this group of hypercalciuric children was no different from that of control children. Suppression of milk and dairy products causes a nutritionally inappropriate diet. PMID- 9214229 TI - [Neutropenic enterocolitis in children with cancer]. AB - OBJECTIVES: Neuropenic enterocolitis (NEC) is a destructive lesion of the ileocecal region occurring in cancer patients treated with chemotherapy. Its clinical picture is one of febrile acute abdominal extension with bloody diarrhea and low neutrophil counts. Our aim was to determine the incidence of NEC in children with cancer and to review the indications of surgery in these cases. MATERIAL AND METHODS: The records of children with cancer treated with chemotherapy in the last 6 years at Hospital Infantile La Paz were reviewed. We selected those patients who had abdominal pain and neutropenia and whose physical examination and radiological findings were consistent with NEC. RESULTS: Twelve cases of NEC were diagnosed during this period among 432 malignancies. The symptoms most frequently seen were abdominal pain and distension, nausea and vomiting. The neutrophil count was consistently below 500/ml. All patients were receiving chemotherapy before the onset of the clinical picture. Five children were operated upon. In three of these we found various ileocecal perforations, in one a gastric perforation and in the remaining one a diffuse inflammation of the ileocecal area. Two non-operated patients died from NEC. The remaining children recovered without problems with medical therapy. CONCLUSION: Pediatric surgeons treating neutropenic cancer patients should be familiar with this condition, that must be suspected early in granulocytopenic patients with acute abdominal extension. Aggressive surgical management is indicated in cases with severe peritonitis, bowel perforation or massive lower gastrointestinal bleeding, irrespective of the degree of neutropenia. Medical support should aim at reestablishing normal neutrophil counts. PMID- 9214230 TI - [Early discharge in neonatology]. PMID- 9214231 TI - [Hypercapnia induced reversible opening of the blood-brain barrier in experimental hyperbilirubinemia]. AB - OBJECTIVE: The incidence of bilirubin encephalopathy in extremely ill neonates with low serum bilirubin is stimulating interest in factors that can modify the blood-brain barrier permeability and allow the passage of bilirubin-albumin macrocomplexes, which may be produced by hypercapnia. MATERIALS AND METHODS: Without modifying the normal bilirubin-albumin affinity, we increased cerebral flow by two levels of hypercapnia (PCO2 67.1 and 101.9 mmHg), acquired by inhalation of CO2 enriched air (5 and 15%) and a decrease in pH (7.00 and 6.93). RESULTS: We demonstrate an increment in cerebral bilirubin deposition (control 2.43 +/- 0.54 mg/g versus 3.29 +/- 0.43 and 3.28 +/- 1.07, F = 4.80, p < 0.05) and I125 albumin (control 26.1 +/- 95.8 micrograms/g versus 364.4 +/- 156.1 and 430.8 +/- 122.4, F03.34, p < 0.05). PMID- 9214232 TI - [Evaluation of three rapid methods for intrapartum detection of group B streptococcus]. AB - OBJECTIVE: The goal of this study was to evaluate three methods for rapid group B streptococcus (GBS) intrapartum vaginal detection. MATERIALS AND METHODS: In 330 women, at risk of delivering an infant with GBS disease, vaginal exudates were collected and a culture performed. The following rapid tests were also performed: 1) Equate Strep B immunoassay in 133 samples. 2) Icon Strep B immunoassay in 192 samples. 3) Co-agglutination with Phadebact Strep B, with a previous incubation (> 4 hours) of the vaginal swabs in Lim Group B Strep broth, in 88 samples. In some patients, two of these methods were performed simultaneously. RESULTS: GBS was detected in 37 women (11.2%) by culture. The sensitivity of Equate Strep B was 47%, Icon Strep B was 35% and co-agglutination with Phadebact Strep B was 38%. The specificity was 91%, 99% and 100% for each one of these methods. PPV 44%, 90% and 100%, respectively and NPV 92%, 91% and 90%, respectively. CONCLUSION: In conclusion, none of these methods was shown sensitive enough to be used for the routine detection of GBS. Therefore, in order to know the GBS carrier status and prevent its vertical transmission, the practice of vaginal culture during late pregnancy is mandatory. PMID- 9214233 TI - [Characteristics of group b streptococcus vertical transmission]. AB - OBJECTIVE: The goal of this study was to compare the characteristics of group B streptococcus (GBS) or Streptococcus agalactiae vertical transmission in woman, receiving or not intrapartum. antimicrobial prophylaxis, at risk of delivering an infant with GRS disease. MATERIALS AND METHODS: The study included 330 women, with risk factors for delivering an infant with GBS disease. The vaginal GBS colonization was studied by conventional culture. A clinical and microbiological follow-up was done in both women and neonates. RESULTS: GHS was detected in 37 women (11.2%). Among these, 21 (56.8%) received intrapartum antibiotics and 16 (43.2%) did not. Forty-one neonates were born from these 37 women. Of these, 11 showed signs of infection (3 with positive blood culture and 8 with blood culture negative for GBS) and 2 presented an asymptomatic bacteremia A GBS neonatal infection (with positive blood culture) was produced in 4.8% of newborns from mothers who received intrapartum antibiotics versus 25% of newborns from mothers who did not receive intrapartum antibiotics. However, this difference was not significant nor was the difference between external colonization by GBS between these two groups of newborns. On the contrary, significant differences were found in the percentage of clinically suspected sepsis (with negative blood cultures), which was more frequent among newborns from mothers without intrapartum antibiotics (30.4% vs 5.6%). A good correlation between the intensity of vaginal colonization and the incidence of microbiologically demonstrated sepsis, suspected sepsis an asymptomatic bacteremia in the newborn was found. CONCLUSION: In conclusion, in order to minimize the vertical transmission of GBS, the most efficient strategy seems to be to offer antibiotic prophylaxis to women identified as GBS carriers, since the antibiotic administration to women with "obstetric risks" often means that it is impossible that two hours elapse between antibiotic administration and delivery, resulting in the loss of efficacy of this second strategy. PMID- 9214234 TI - [Kawasaki disease with early bilateral coronary aneurysm]. PMID- 9214235 TI - [Toxic effects of the second dose of sodium aurothiomalate in childhood systemic chronic arthritis]. PMID- 9214236 TI - [Clinical aspects and therapeutic follow-up of a girl with lumbosacral teratoma]. PMID- 9214237 TI - [Emryopathy due to isotretinoine. A case history of one observation]. PMID- 9214239 TI - [Intramedullary cavernous angioma in a 10-year-old child: a case report]. PMID- 9214238 TI - [Hypophosphatasia in childhood]. PMID- 9214240 TI - [Gaucher's disease: results of enzymatic treatment with beta glucosidase acid]. PMID- 9214241 TI - [A girl with pancytopenia, short stature and minor skeletal abnormalities]. PMID- 9214242 TI - [Several considerations about immunological reconstruction]. PMID- 9214243 TI - [Spontaneous pneumopericardium in a neonate without other anomalies]. PMID- 9214244 TI - [Acoustic reflexometry in newborn]. PMID- 9214245 TI - [Clavicular fracture in a neonate]. PMID- 9214246 TI - Patient preferences for communication with physicians about end-of-life decisions. SUPPORT Investigators. Study to Understand Prognoses and Preference for Outcomes and Risks of Treatment. AB - BACKGROUND: Physicians are frequently unaware of patient preferences for end-of life care. Identifying and exploring barriers to patient-physician communication about end-of-life issues may help guide physicians and their patients toward more effective discussions. OBJECTIVE: To examine correlates and associated outcomes of patient communication and patient preferences for communication with physicians about cardiopulmonary resuscitation and prolonged mechanical ventilation. DESIGN: Prospective cohort study. SETTING: Five tertiary care hospitals. PATIENTS: 1832 (85%) of 2162 eligible patients completed interviews. MEASUREMENTS: Surveys of patient characteristics and preferences for end-of-life care; perceptions of prognosis, decision making, and quality of life; and patient preferences for communication with physicians about end-of-life decisions. RESULTS: Fewer than one fourth (23%) of seriously ill patients had discussed preferences for cardiopulmonary resuscitation with their physicians. Of patients who had not discussed their preferences for resuscitation, 58% were not interested in doing so. Of patients who had not discussed and did not want to discuss their preferences, 25% did not want resuscitation. In multivariable analyses, patient factors independently associated with not wanting to discuss preferences for cardiopulmonary resuscitation included being of an ethnicity other than black (adjusted odds ratio [OR], 1.48 [95% CI, 1.10 to 1.99), not having an advance directive (OR, 1.35 [CI, 1.04 to 1.76]), estimating an excellent prognosis (OR, 1.72 [CI, 1.32 to 2.59]), reporting fair to excellent quality of life (OR, 1.36 [CI, 1.05 to 1.76]), and not desiring active involvement in medical decisions (OR, 1.33 [CI, 1.07 to 1.65]). Factors independently associated with wanting to discuss preferences for resuscitation but not doing so included being black (OR, 1.53 [CI, 1.11 to 2.11]) and being younger (OR, 1.14 per 10-year interval younger [CI, 1.04 to 1.25]). CONCLUSIONS: Among seriously ill hospitalized adults, communication about preferences for cardiopulmonary resuscitation is uncommon. A majority of patients who have not discussed preferences for end-of-life care do not want to do so. For patients who do not want to discuss their preferences, as well as patients with an unmet need for such discussions, failure to discuss preferences for cardiopulmonary resuscitation and mechanical ventilation may result in unwanted interventions. PMID- 9214247 TI - Predictive power of duplex ultrasonography in asymptomatic carotid disease. AB - BACKGROUND: Duplex ultrasonography is considered a valid measure of stenosis of the carotid arteries, but the prognostic value of repeated ultrasonographic examinations is unknown. OBJECTIVE: To determine the ability of serial ultrasonographic measurements to predict cerebrovascular events in patients with asymptomatic carotid disease. DESIGN: Secondary analysis of data from a natural history study of asymptomatic carotid disease. PATIENTS: Asymptomatic patients with cervical bruits. MEASUREMENTS: Duplex ultrasonography of the carotid arteries was done at study enrollment and biannually thereafter. Multivariable Cox proportional hazards models with fixed and time-dependent covariates were used for analysis. RESULTS: 61 transient ischemic attacks (TIAs) and 38 strokes occurred in 715 participants over a mean follow-up period of 3.2 years; 4 strokes were disabling, and no deaths from stroke occurred. Sixty percent of strokes occurred in persons who did not have severe stenosis. One fifth of participants had stenosis progression. Baseline carotid stenosis was a significant predictor of the outcome "TIA or stroke" (relative risk, 1.5 [95% CI, 1.2 to 1.7]) and retained its predictive ability for more than 3 years. Progression of stenosis to 80% or more significantly increased the risk for cerebrovascular events and death. The sensitivity and positive predictive value of progression as an independent predictor of TIA or stroke were low. CONCLUSION: Severe carotid stenosis is associated with a higher risk for cerebrovascular events, but the power of repeated ultrasonography to predict ischemic events is limited by low incidence rates and low rates of progression. The evidence does not support the routine use of serial ultrasonography to determine the risk for stroke in unselected patients with asymptomatic carotid disease. PMID- 9214248 TI - Detection of the His1069Gln mutation in Wilson disease by rapid polymerase chain reaction. AB - BACKGROUND: Most known mutations in the gene associated with Wilson disease are rare. Only the His1069Gln mutation is found often in patients of Northern or Eastern European origin. OBJECTIVE: To examine the frequency of the His1069Gln mutation in Austrian patients with Wilson disease and their families by using a new, rapid polymerase chain reaction (PCR) test. DESIGN: Cross-sectional study. SETTING: University medical center. PATIENTS: 83 patients from 72 families and 98 relatives of 11 homozygous index patients. MEASUREMENTS: Results of a semi-nested PCR-based assay to detect the His1069Gln mutation in Wilson disease, clinical symptoms, and liver histologic findings. RESULTS: 20 patients, including 5 siblings, were homozygous for the His1069Gln mutation. Thirty-three patients, including 4 siblings, were compound heterozygotes. The mutation was not detected in 30 patients, including 2 siblings. Homozygotes were older at onset of symptoms (mean age, 24 +/- 6 years) than compound heterozygotes (17 +/- 6 years [95% CI, 3.3 to 10.7 years]; P = 0.0135) and patients with other mutations (18 +/- 8 years [CI, 1.8 to 10.2 years]; P = 0.117). Homozygotes were more often female (73.3%) than were compound heterozygotes (48% [CI, 0.94% to 2.46%]) and patients with other mutations (50% [CI, 0.91% to 2.37%]) (P = 0.05). Four of 98 asymptomatic relatives of 11 homozygous index patients were also homozygotes. Heterozygosity was confirmed in 46 relatives (19 parents, 11 children, and 16 distant relatives). CONCLUSION: The His1069Gln mutation was detected in 61% of Austrian patients with Wilson disease. Polymerase chain reaction may be useful for diagnosis and screening of family members of homozygous index patients, even if first-degree relatives are not available for examination. PMID- 9214249 TI - Proximal bursitis in active polymyalgia rheumatica. AB - BACKGROUND: The cause of musculoskeletal symptoms in the proximal extremities of patients who have polymyalgia rheumatica is not completely understood. The diffuse and severe discomfort can only be partially explained by the mild joint synovitis that is observed in these patients. OBJECTIVE: To determine the involvement of the synovial structures of the shoulder girdle of patients who have active symptoms of polymyalgia rheumatica. DESIGN: Case-control study. SETTING: 2 secondary referral centers of rheumatology. PATIENTS: 13 case-patients who had active symptoms of polymyalgia rheumatica seen during a 6-month period, 9 control-patients who had early symptoms of elderly-onset rheumatoid arthritis, and 10 age-matched healthy controls. MEASUREMENTS: Magnetic resonance imaging of the shoulder was done on the 13 case-patients, 9 control-patients, and 10 healthy controls. RESULTS: The frequency of subacromial and subdeltoid bursitis was significantly higher in the case-patients (who had polymyalgia rheumatica) than in the control-patients (who had elderly-onset rheumatoid arthritis). The frequencies of synovitis of the joints and tenosynovitis of the biceps did not significantly differ between the 13 case-patients and the 9 control-patients. None of the healthy controls showed evidence of fluid accumulation in the joints, bursae, or sheaths of the long head of the biceps. CONCLUSIONS: Inflammation of subacromial and subdeltoid bursae in association with synovitis of the glenohumeral joints and tenosynovitis of the biceps may contribute to the diffuse discomfort in the shoulder girdle observed in patients with polymyalgia rheumatica. PMID- 9214251 TI - How consumers and policymakers can use systematic reviews for decision making. AB - Systematic reviews can be a very useful decision-making tool because they objectively summarize large amounts of information, identify gaps in medical research, and identify beneficial or harmful interventions. Consumers can use systematic reviews to help them make health care decisions. Policymakers can use systematic reviews to help them make decisions about what types of health care to provide. Despite the potential value of systematic reviews, little evidence of their direct impact on the decisions made by consumers and policymakers is available. We discuss strategies for optimizing the use of systematic reviews by increasing the awareness and identification of reviews, learning to critically evaluate the findings of reviews, and overcoming barriers to the incorporation of reviews into the decision-making process. In addition, the participation of consumers and policymakers in the design, conduct, and reporting of systematic reviews can help to produce reviews that are relevant and understandable to target audiences. Because decisions that involve health care policies and issues are complex processes in which information (such as that provided by systematic reviews) plays only a part, strategies for increasing the use of systematic reviews should be evaluated for their usefulness in the decision-making process. PMID- 9214250 TI - A nosocomial outbreak of multidrug-resistant tuberculosis. AB - BACKGROUND: An outbreak of seven cases (in six patients and one health care worker, all of whom had AIDS) of multidrug-resistant tuberculosis occurred in a hospital in Chicago. The hospital had a respirator-fit testing program but no acid-fast bacilli isolation rooms. OBJECTIVE: To identify risk factors for transmission of Mycobacterium tuberculosis. DESIGN: Retrospective cohort study. SETTING: Private hospital. PARTICIPANTS: Patients and health care workers exposed to M. tuberculosis. MEASUREMENTS: Analysis of M. tuberculosis isolates, tuberculin skin testing, assessment of exposure, and assessment of participant characteristics. RESULTS: All seven M. tuberculosis isolates had matching DNA fingerprints. Of patients exposed to M. tuberculosis, those who developed tuberculosis had lower CD4+ T-lymphocyte counts (P = 0.02) and were more likely to be ambulatory (P = 0.03) than those who did not. Of 74 exposed health care workers, the 11 (15%) who had conversion on tuberculin skin testing were no more likely than those who did not have conversion to report that they always wore a respirator with a high-efficiency particulate air filter. CONCLUSIONS: Transmission of M. tuberculosis occurred in a hospital that did not have recommended isolation rooms. A respirator-fit testing program did not protect health care workers in this setting. PMID- 9214252 TI - Update in general internal medicine. PMID- 9214253 TI - Costs, outcomes, and patient satisfaction by provider type for patients with rheumatic and musculoskeletal conditions: a critical review of the literature and proposed methodologic standards. AB - PURPOSE: To compare the outcomes of care provided by generalists with that provided by specialists for patients with musculoskeletal and rheumatic conditions. DATA SOURCES: English-language studies published between 1986 and April 1996 were identified through a MEDLINE search. STUDY SELECTION: Studies that compared generalists' and specialists' treatment preferences, appropriateness of care, or outcomes with regard to musculoskeletal and rheumatic conditions were examined. DATA EXTRACTION: Studies were reviewed for methodologic rigor and outcomes. DATA SYNTHESIS: Low back pain is treated by many types of providers, without consistent differences in outcomes across provider types. In one study, however, patients were more satisfied with chiropractic care than with care provided by primary care physicians, although the former cost twice as much as the latter. For osteoarthritis of the hip, rheumatologists and primary care providers reported using different therapeutic regimens. For acute mono- and oligoarthritis, rheumatologists performed arthrocentesis more appropriately than nonrheumatologists and produced shorter durations of hospitalization. In the management of gout, rheumatologists used colchicine during the introduction of urate-lowering therapy more appropriately than other providers. In two population based cohorts of patients with rheumatoid arthritis, patients cared for by rheumatologists were prescribed significantly more disease-modifying agents and had less disability than patients cared for by generalists. CONCLUSIONS: Although empirical data are scant, there seem to be differences between generalists and specialists for a range of outcomes in various musculoskeletal and rheumatic conditions. Studies to data have important methodologic limitations that need to be addressed in future research. PMID- 9214254 TI - Advising patients who seek alternative medical therapies. AB - Alternative medical therapies, such as chiropractic, acupuncture, homeopathy, and herbal remedies, are in great public demand. Some managed care organizations now offer these therapies as an "expanded benefit." Because the safety and efficacy of these practices remain largely unknown, advising patients who use or seek alternative treatments presents a professional challenge. A step-by-step strategy is proposed whereby conventionally trained medical providers and their patients can proactively discuss the use or avoidance of alternative therapies. This strategy involves a formal discussion of patients' preferences and expectations, the maintenance of symptom diaries, and follow-up visits to monitor for potentially harmful situations. In the absence of professional medical and legal guidelines, the proposed management plan emphasizes patient safety, the need for documentation in the patient record, and the importance of shared decision making. PMID- 9214255 TI - Genetic heterogeneity in Wilson disease: lessons from rare alleles. PMID- 9214256 TI - Building the best team. PMID- 9214257 TI - Annals' 70th anniversary: a look back and a look ahead. PMID- 9214258 TI - Diagnosing syncope. Part 2: Unexplained syncope. Clinical Efficacy Assessment Project of the American College of Physicians. AB - PURPOSE: To review the literature on diagnostic testing in syncope that remains unexplained after initial clinical assessment. DATA SOURCES: MEDLINE search. STUDY SELECTION: Published papers were selected if they addressed diagnostic testing in syncope, near syncope, or dizziness. DATA EXTRACTION: Studies were identified as population studies, referral studies, or case series. DATA SYNTHESIS: After a thorough history, physical examination, and electrocardiography, the cause of syncope remains undiagnosed in 50% of patients. In such patients, information may be derived from the results of carefully selected diagnostic tests, especially 1) electrophysiologic studies in patients with organic heart disease, 2) Holter monitoring or telemetry in patients known to have or suspected of having heart disease, 3) loop monitoring in patients with frequent events and normal hearts, 4) psychiatric evaluation in patients with frequent events and no injury, and 5) tilt-table testing in patients who have infrequent events or in whom vasovagal syncope is suspected. Hospitalization is indicated for high-risk patients, especially those with known heart disease and elderly patients. CONCLUSIONS: A flexible, focused approach is required to diagnose syncope. Features of the initial history and physical examination help guide diagnostic testing. PMID- 9214259 TI - New types of cancer after basal-cell cancer. PMID- 9214260 TI - New types of cancer after basal-cell cancer. PMID- 9214261 TI - Predicting death in mechanically ventilated recipients of bone marrow transplants. PMID- 9214262 TI - Thyrotoxicosis insistiates: report of 17 cases. PMID- 9214263 TI - Hepatitis C virus genotype, hepatitis C virus RNA titers, and response to interferon. PMID- 9214264 TI - Human granulocytic ehrlichiosis: a cardiac risk factor? PMID- 9214265 TI - Prevention of corticosteroid-induced osteoporosis. PMID- 9214267 TI - [Nuclear medicine approach for the detection and characterization of liver metastasis]. AB - Nuclear Medicine procedures include in vitro techniques able to detect and characterize liver metastases. Among the in vitro applications, circulating tumour markers level determination provides information about the response to therapy and the presence of active disease. While after a successful antineoplastic treatment the serum biological marker level usually falls, a progressive increase represents an alarm signal of occult/residual tumour or metastatic dissemination. However, this method is not able to identify the site of the lesion, therefore complementary imaging techniques are needed to confirm or exclude the diagnostic suspicion. The main advantages of the in vitro methods are the low cost and, consequently, the possibility of periodically checking of tumour activity. The in vivo nuclear medicine techniques are usually performed as second level test. In fact, they can play a role in detecting and characterizing hepatic metastases when radiological methods are inconclusive or when selected tumour seeking agents (i.e. radiolabelled antibodies, radioiodinated mIBG, Octreotide, etc.) can be used. Moreover, the introduction of PET procedures has provided physicians with a useful tool for the evaluation of tumour behaviour (glucose consumption, proteic, synthesis, receptor expression). A further significant diagnostic improvement is represented by the recent introduction of the multimodality co-registration of images (including CT scan, MRI, SPET, PET). The fusion imaging allows a superimposition of the functional and metabolic information to the morphological one. PMID- 9214266 TI - [Diagnostic imaging of liver metastasis]. AB - The early diagnosis and monitoring of hepatic metastases are now achieved by different imaging modalities, some using ionizing radiations (computed tomography and angiography), some based on other energy sources (sonography and magnetic resonance), but all coming within the radiological area, which offers concrete possibilities of integration and the necessary organization. These modalities are sometimes used only for percutaneous histological samplings with minimal invasiveness. The progress in hepatic resective surgery and the possibility of orthotopic liver transplantation for some neoplastic histotypes, together with the alternatives provided by interventional radiology, have brought a continuous updating of the specialist' interest in the morphological and functional definition of hepatic metastatic disease, with the specific aim of choosing the best therapeutic strategy. Hepatic metastases have the greatest impact on the survival of patients with gastrointestinal neoplasms, especially colonic adenocarcinoma. Intraoperative sonography and CT arterial portography currently provide greatest diagnostic sensitivity in terms of spatial resolution but cannot be considered as methods of choice, the former for obvious reasons and the latter because of its invasiveness and complexity. The alternatives are to be sought in spiral CT and the new MR sequences which can undoubtedly provide a decisive improvement in the diagnostic standards currently available. Profoundly changed, but no less important, is the role of angiography, which still provides the anatomical support for hepatic surgery and the means for alternative treatments, such as chemoembolization and continuous infusional chemotherapy. PMID- 9214268 TI - [Liver metastasis. Clinico-pathological prognostic factors in metastasis from colorectal cancer]. AB - Hepatic Metastases (HM) from colorectal cancer represents one of the main problems of oncologic treatment: from 80 to 90 percent of patients undergo chemotherapy and a minority hepatic resection. The natural history of patients with unresectable HM has been recently investigated by uni and multivariate analyses; the percentage of hepatic replacement, the stage and grade of primary colorectal tumours, alkaline phosphatase and the presence of extrahepatic disease proved to be the most important independent prognostic factors. Albumin and carcino-embryonic antigen (CEA) levels, age and weight loss of patients were also prognostic. The groups of patients with more favourable factors had a median survival ranging from 21 to 35 months, in contrast to a median survival of 4 to 8 months for those with adverse factors. The outcome of more than 3400 patients submitted to hepatic resection, has been investigated. At multivariate analysis twelve variables resulted independently related to prognosis: stage of primary tumour, extent of liver involvement and presence of extrahepatic metastases were considered to be the most important. The knowledge of prognostic factors is extremely important in selecting patients candidated to various treatments, to interpret the results and to plan new therapeutic strategies. PMID- 9214269 TI - [Liver metastasis: therapeutic strategy]. AB - The liver is one of the most frequent sites of metastatic growth, in particular from digestive malignancies (DM). The first goal is to reduce the incidence of metastases. Adjuvant systemic chemotherapies have been demonstrated to reduce the recurrence rate and to improve survival in Dukes C colon cancer. Fluorouracil is the pivot of adjuvant treatment modulated by Leucovorin or Levamisol. A short postoperative administration of fluorouracil by intraportal route has been tested, but the results are controversial. Adjuvant treatments for different DM are under investigation. When hepatic metastases are clinically evident, therapeutic decisions depend on several factors: site and nature of primary, extent of hepatic and extrahepatic disease, patient characteristics, efficacy of treatments. A staging system should be adopted to allow a rational approach. In selected cases a locoregional treatment can achieve consistent results. Hepatic Intrarterial Chemotherapy (HIAC) for colorectal metastases achieves objective responses in more than 50% of patients. Survival seems positively affected. When feasible, Ro hepatic resection is the most effective treatment, five-year survival rate being about 30% when metastases are from colorectal cancer. Since the liver is the most frequent site of recurrence after resection, repeat resection have been successfully performed. PMID- 9214270 TI - [Locoregional chemotherapy of liver metastasis from colorectal carcinoma]. AB - Hepatic metastases are a major cause of death in patients with colorectal carcinoma. Traditional intravenous chemotherapy produces responses in 10% to 30% of patients and surgical resection is feasible in approximately 20% of patients. Infusion of cytotoxic agents into the hepatic artery is the most promising form of therapy for unresectable hepatic metastases. The recent development of a totally implantable pump has allowed prolonged infusion of chemotherapeutic agents with a good compliance and quality of life of the patients. The rationale for hepatic arterial infusion (HAI) present an anatomical and pharmacological basis with the use of agents with high hepatic extraction resulting in minimal systemic toxicity. The results of eight randomized trial assessing the value of HAI Floxuridine shows that such regional chemotherapy increases the likelihood of hepatic response compared with systemic treatment (52% vs 15%). Survival information is difficult to evaluate because some of the studies are small, some had a crossover design and some others had bias factors. Extrahepatic disease develops in 40-70% of patients undergoing HAI; the use of systemic therapy plus HAI may produce a decrease in extrahepatic disease. Further studies of combined systemic/arterial regiment are necessary. PMID- 9214271 TI - [Physical and chemical locoregional therapy in liver metastasis]. AB - Close imaging follow-up of patients with known colorectal adenocarcinoma and other cancer can often detect hepatic metastases that are still small and potentially treatable. Unfortunately, many of these patients are poor surgical candidates, or refuse operation. This has stimulated the search for other minimally invasive means of tumor ablation. The first procedure suggested for this type of treatment was percutaneous ultrasound guided ethanol injection. Complete response is often obtained in lesions less than 2-3 cm, but only partial response in greater lesions. Usually a decrease of CEA values is observed, lasting for some months. Clearly, is difficult to establish whether, how much, and in which cases patient survival is lengthened. In the same time, because the procedure has proved safe, simple and cheap, when palliative purpose is requested it is hard to deny treatment to an inoperable patient with not advanced disease. Response in endocrine metastases in higher, because of their hypervascularity. Recently, encouraging results have been obtained inducing coagulation necrosis by means of thermal methods as laser or radiofrequency. In particular, new strategies of the latter obtained very promising results. PMID- 9214272 TI - [Adjuvant chemotherapy following liver metastasis resection in neoplasms of the colorectum]. AB - INTRODUCTION: Adjuvant chemotherapy after hepatic resection of colorectal metastases has been advocated because of the significant risk of tumor recurrence. Data presented in literature show unequivocally that the probability of recurrence and tumor-related death exceeds survival. Although the initial site of disease recurrence varies, both intrahepatic and extrahepatic organs are at risk for tumor progression. PATIENTS AND METHODS: To reduce the risk of tumor progression after hepatic resection of colorectal metastases, multiple chemotherapy approaches have been designed. Treatment approaches have varied by chemotherapeutic agents, route and schedule of administration, and duration. RESULTS: A significant difference in survival between patients treated by resection alone and resection with adjuvant chemotherapy is not evident from these data. DISCUSSION: Metastatic inefficiency is a major driving force in the development of metastatic patterns. Then, the probability of synchronous seeding (from primary colorectal carcinomas) of hematogenous metastases in the liver, lungs, and other organs is less than metachronous seeding, in which only liver metastases are predominantly seeded from the hepatic metastases, and arterial metastases are predominantly seeded from the pulmonary lesions. This sequential development of metastatic patterns creates windows of potential therapeutic opportunity during which metastatic disease is confined to the liver alone. CONCLUSION: The risk of recurrence after hepatic resection of colorectal metastases clearly warrants further prospective study of adjuvant therapy to improve survival. Future trials of adjuvant therapy after hepatic resection of colorectal metastases must address the patterns of tumor recurrence and account for adjuvant chemotherapy given previously, after resection on the primary colorectal carcinoma. PMID- 9214274 TI - [Colorectal carcinoma: therapeutic approach in patients already treated with metastasis resection]. AB - INTRODUCTION: Fluorouracil is the most wildly used agent in the treatment of advanced colorectal cancer after mastectomy, with an overall response rate of 20%. To improve response rate and survival new therapeutic approaches have been designed such as biochemical modulation of fluorouracil, new schedules of infusion and new drugs. PATIENTS AND METHODS: Many drugs have been used for biochemical modulation of fluorouracil, and leucovorin is very effective. New schedules of infusion such as continuous i.v. infusion of fluorouracil and new drugs like doxifluridine and irinotecan are promising. DISCUSSION: Literature data suggest that biochemical modulation of fluorouracil by lederfolin is effective in the treatment of advanced colorectal cancer such as continuous i.v. infusion of fluorouracil and new drugs, especially doxifluridine, may improve therapeutic effects. CONCLUSION: Chemotherapy with fluorouracil remain the treatment of choice of patients with colorectal cancer. Further studies are necessary to identify the role of new biochemical modulation, new schedule and new drugs. PMID- 9214273 TI - [Clinical features and symptomatic treatment of liver metastasis in the terminally ill patient]. AB - The incidence of liver metastasis is quite frequent in patients with advanced cancer. Some patients are asintomatic, but more often a correlation can be present between the clinical observation and the anatomic and functional alteration of the liver provoked by metastasis. Hepatomegaly may cause pain, dyspnea, hiccup. Biliary obstruction generates jaundice and itching. Portal hypertension may cause ascitis, encephalopathy, varices of the esophagus. Hepatic failure may produce symptoms like sopor, dysrasic oedema, coagulation problems, jaundice. The treatment of the symptoms listed above is crucial for the quality of life of these patients, and must be the mainstay of the therapeutic approach. This paper describes the palliative treatment of the clinical complications related to liver metastasis. PMID- 9214275 TI - [Surgical treatment of pulmonary metastasis from breast carcinoma. Personal contribution and considerations on the experience in the literature]. AB - Considering a series of twelve patients operated on at the General Surgical Clinic of the University of Pavia, the authors discuss the results of surgical therapy of pulmonary metastases from breast cancer. According to literature data their results are not so good with a 5-year survival rate of 11%. Nevertheless, selected series of patients have been reported with a 5-year survival rate of 43% after pulmonary resection. So, by a careful selection of the indications the possibility of a surgical treatment would not be eliminated. After breast cancer exeresis it is certain that surgery is the best treatment for a solitary pulmonary nodule when there is some doubt about the diagnosis of primary or secondary lung cancer. PMID- 9214276 TI - [Complications of surgery of Dupuytren disease. Comparison of total and partial aponeurectomy]. AB - Total and partial aponeurectomy are the most usual operations performed for the treatment of Dupuytren's disease. Eighteen patients, for a sum of 23 treated hands, were submitted to total aponeurectomy in 20 cases and to partial aponeurectomy in 3 cases. Recurrence was present in 4 cases. Postoperative morbidity was 50% (10/20 cases) following total aponeurectomy and 0% (0/3 cases) following partial aponeurectomy. In patient with recurrence disease postoperative complications were present in 3/4 cases (75%) while in patients, operated for the first time, morbidity rate was 36.8% (7/19 cases). Postoperative complications following total aponeurectomy are extremely frequent, especially if recurrence is present. Partial aponeurectomy allows to achieve functional result as good as total aponeurectomy with significative reduction of postoperative morbidity. PMID- 9214277 TI - [Polymer biomaterials (polyphosphazenes) in the repair of peripheral nervous system]. AB - Biodegradable polymers gives interesting perspectives of use in making artificial conduits for peripheral nerve reconstruction. Poliphosphazenes are materials highly biocompatible and have a controllable reabsorption rate. According to the substitutes that are introduced in the molecule, they can also be used as a framework for drug release. Conduits obtained with poli [bis(etilalanate) phosphazene] were evaluated as guides for nerve regeneration in an experimental animal model. In six Wistar rats, under general anesthesia and with microsurgical technique, the ischiatic nerve was isolated. On the right side a segment of the nerve was removed in order to create a 10 mm gap. The defect was then repaired using the conduit. On the controlateral limb the nerve continuity was restored using as an autograft the segment removed from the right. Control were performed at 30, 90, 180 days and consisted in histological and electron microscopy investigations. They showed the gradual degradation of the conduit without signs of local and general toxicity. The regeneration of the nerve fibers in the lumen of the conduit was not significantly different from the one obtained with the autologous grafts. So poliphosphazene conduits may be considered effective as a guide for nerve regeneration, above all for the possibility of use the polymer as a carrier for neurite-promoting factors. PMID- 9214278 TI - Agonist potency ratios at m1 and m3 muscarinic receptors expressed in A9L and CHO cells. PMID- 9214279 TI - Nutritional Implications of Macronutrient Substitutes. Arlington, Virginia, October 27-29, 1996. Conference proceedings. PMID- 9214280 TI - [The sensitivity of Pseudomonas mallei strains to antibacterial preparations in in-vitro experiments on a cell culture model]. AB - The cytopathogenic action of P. mallei on macrophages of golden hamsters and transplantable Hep-2 cell culture was studied. It was shown that the culture test systems may be efficient in estimation of the P. mallei virulent properties. The effect of antibacterial drugs on intracellular P. mallei was also studied. PMID- 9214281 TI - [A chronobiological study of the function of the epithelial proliferative system of the murine esophagus after its infection with typhoid and the use of lomefloxacin]. AB - The rhythmical character of the function of the proliferative system of the oesophagus epithelium of mice changed after infection by Salmonella typhi and after injection of lomefloxacin to intact and infected mice. The degree of the changes depended on the exposure time. The proliferative system of the oesophagus epithelium was more resistant to the infection in the night-time than the day time and equally resistant to the effect of lomefloxacin in both the day-time and the night-time. The proliferative system of the oesophagus epithelium of the infected mice exposed to lomefloxacin in the day-time proved to be in a noncompensated state and in the night-time it proved to be in the hyperfunctional state. PMID- 9214282 TI - [Enhancement of treatment efficacy in experimental plague by the sequential administration of antibiotics]. AB - It was shown that the use of ampicillin, azlocillin or polymyxin 24 or 96 hours after the plague infection at the background of the every-day use of rifampicin in the doses protecting only 30 per cent of the animals from death provided 80 100-percent survival of the animals. With the every-day use of ampicillin, azlocillin or polymyxin in succession with rifampicin there was observed a 3-fold increases in the survival of the albino mice as compared to those exposed to an analogous dose of rifampicin alone. A decrease in the number of administrations of the above drugs and an increase in the intervals between the administration also resulted in a significant rise of the animal survival in comparison with that after the every-day use of a similar dose of rifampicin. PMID- 9214284 TI - [The growth of Rhodotorula rubra yeasts and their synthesis of ergosterol on media with lovastatin]. AB - The effect of lovastatin, a competing inhibitor of 3-hydroxy-3-methylglutaryl-CoA reductase (HMG-CoA-reductase) of the fungal origin on the growth of and ergosterol biosynthesis by Rh. rubra VKPM Y 1337 was studied. It was shown that the yeast growth was inhibited by lovastatin in concentrations of 0.25 to 5.0 micrograms/ml when the inhibitor was added to the cultivation medium either with the inoculum or at later periods of the yeast cultivation. In concentrations of 2.0 to 5.0 micrograms/ml lovastatin almost completely inhibited the yeast growth. In the concentrations tested the inhibitor did not decrease the ergosterol level in the yeast cells below 40 per cent of the control. When lovastatin was added in concentrations of 1.0 to 5.0 micrograms/ml the maximum inhibition of the ergosterol biosynthesis was observed at the end of the growth log phase. Further cultivation of the yeast in the presence of the inhibitor resulted in an increase of the ergosterol biosynthesis. It is believed that the increase in the quantity of ergosterol may be due either to the HMG-CoA-reductase induction under the action of the inhibitor or to its inactivation as a result of the hydroxylation. PMID- 9214283 TI - [The efficacy of the eubiotic bifacide when used in the combined antibacterial therapy of newborn infants with infectious-inflammatory diseases]. AB - The efficacies of bifacid and bifidumbacterin were studied comparatively in the correction of intestinal biocenosis in 60 newborns with infectious inflammatory diseases and intestine disfunction treated with massive doses of antibacterial drugs. The study showed that the use of bifidumbacterin was accompanied by significant disturbances in the biocenosis and by development of the intestinal syndrome. The protective action of the drug was observed after a short-term use only of one antibiotic and when the course of the bifidum therapy was continued after discontinuation of the treatment with antibacterial drugs. The use of bifacid was accompanied by a rapid (by the 2nd or the 5th day fo the treatment) and stable normalization of the stools and a marked improvement of the patient general state. The clinical efficacy of bifacid was much higher than that of bifidumbacterin. At the background of the bifacid therapy there was observed correction of the intestinal microflora composition due to normalization of the count of Bifidobacterium, Lactobacillus and Colibacillus as well as to eradication of opportunistic pathogens. PMID- 9214285 TI - [Oral cephalosporins in the treatment of suppurative-inflammatory diseases]. PMID- 9214286 TI - [The effect of solar ultraviolet on the system of microorganisms-higher plant antibiotics, a natural phenomenon and a new approach to enhancing the efficacy of antibiotics]. AB - Under the effect of solar radiation some antibiotics of plant origin (phenylheptatriin, bakuchiol and others) showed antimicrobial phototoxicity differing by the spectrum and activity from the antibiotic action. The highest in vitro antimicrobial phototoxicity was observed with polyacetylene phenylheptatriin: the activation effect of solar radiation on it was due to UV-A and developed in gaseous phase or to a lesser extent in dispersed liquid phase. For comparison, 18 currently used antibiotics of various chemical structure were investigated and no phototoxicity under the effect of solar radiation with respect to the tested microbes was detected. In nature the phenomenon of antimicrobial phototoxicity of plant secondary metabolites due to the effect of solar radiation is probably of large scale. The study of the phenomenon is a new trend in biology (plant antibiotics and phytoncides, phytoimmunity, ecological and evolutionary microbiology, etc.) and a new approach to increase the efficacy of some antibiotics and to develop principally novel photochemotherapeutics for the treatment of infections in humans, animals and plants. PMID- 9214287 TI - Comparison of NH exchange and circular dichroism as techniques for measuring the parameters of the helix-coil transition in peptides. AB - Circular dichroism and NH exchange are compared directly as techniques for measuring helix content in peptides and the parameters of the helix-coil transition. To cover a broad range of helix contents, alanine-based peptides with chain lengths varying from 12 to 22 residues are examined over the temperature range from 0.6 to 26.9 degrees C in 1 M sodium chloride, 2H2O. Helix-coil transition theory independently fits both circular dichroism and exchange data, but the helix contents measured by exchange are larger than those measured by circular dichroism. The two techniques are brought into agreement by removing the assumption that the intrinsic chemical exchange rate in the helix is the same as the exchange rate measured for short unstructured model peptides. This modification allows the circular dichroism and NH exchange data to be described by the same set of helix parameters and indicates that the intrinsic exchange rate in the presence of helical structure is reduced approximately 17% relative to the rates measured in unstructured models. To test the possibility that this effect is electrostatic in origin, the sensitivity of the exchange reaction to ionic strength is determined. A substantial dependence of exchange rate on ionic strength is found, but the form of the dependence is complex. In studies of the exchange rates of native proteins, the exchange-competent form of the protein is assumed to exchange with the same rate constant as a blocked dipeptide with the identical amino acid sequences. Our result suggests that this assumption will be seriously in error in some cases because of charge effects in the protein. PMID- 9214288 TI - Role of protein-protein interactions in the function of replication protein A (RPA): RPA modulates the activity of DNA polymerase alpha by multiple mechanisms. AB - Replication Protein A (RPA) from human cells is a stable complex of 70-, 32-, and 14-kDa subunits that is required for multiple processes in DNA metabolism. RPA binds with high affinity to single-stranded DNA and interacts with multiple proteins, including proteins required for the initiation of SV40 DNA replication, DNA polymerase alpha and SV40 large T antigen. We have used a series of mutant derivatives of RPA to map the regions of RPA required for specific protein protein interactions and have examined the roles of these interactions in DNA replication. T antigen, DNA polymerase alpha and the activation domain of VP16 all have overlapping sites of interaction in the N-terminal half (residues 1-327) of the 70-kDa subunit of RPA. In addition, the interaction site for DNA polymerase alpha is composed of two functionally distinct regions, one (residues 1- approximately 170) which stimulates polymerase activity and a second (residues approximately 170-327) which increases polymerase processivity. In the latter, both the direct protein-protein interaction and ssDNA-binding activities of RPA were needed for RPA to modulate polymerase processivity. We also found that SV40 T antigen inhibited the ability of RPA to increase processivity of DNA polymerase alpha, suggesting that this activity of RPA may be important for elongation but not during the initiation of DNA replication. DNA polymerase alpha, but not T antigen also interacted with the 32- and/or 14-kDa subunits of RPA, but these interactions did not seem to effect polymerase activity. PMID- 9214289 TI - Structure of human isovaleryl-CoA dehydrogenase at 2.6 A resolution: structural basis for substrate specificity,. AB - Isovaleryl-CoA dehydrogenase (IVD) belongs to an important flavoprotein family of acyl-CoA dehydrogenases that catalyze the alpha,beta-dehydrogenation of their various thioester substrates. Although enzymes from this family share similar sequences, catalytic mechanisms, and structural properties, the position of the catalytic base in the primary sequence is not conserved. E376 has been confirmed to be the catalytic base in medium-chain (MCAD) and short-chain acyl-CoA dehydrogenases and is conserved in all members of the acyl-CoA dehydrogenase family except for IVD and long-chain acyl-CoA dehydrogenase. To understand this dichotomy and to gain a better understanding of the factors important in determining substrate specificity in this enzyme family, the three-dimensional structure of human IVD has been determined. Human IVD expressed in Escherichia coli crystallizes in the orthorhombic space group P212121 with unit cell parameters a = 94.0 A, b = 97.7 A, and c = 181.7 A. The structure of IVD was solved at 2.6 A resolution by the molecular replacement method and was refined to an R-factor of 20.7% with an Rfree of 28.8%. The overall polypeptide fold of IVD is similar to that of other members of this family for which structural data are available. The tightly bound ligand found in the active site of the structure of IVD is consistent with that of CoA persulfide. The identity of the catalytic base was confirmed to be E254, in agreement with previous molecular modeling and mutagenesis studies. The location of the catalytic residue together with a glycine at position 374, which is a tyrosine in all other members of the acyl-CoA dehydrogenase family, is important for conferring branched-chain substrate specificity to IVD. PMID- 9214290 TI - Phenotypically selected mutations in myosin's actin binding domain demonstrate intermolecular contacts important for motor function. AB - Here, we biochemically characterize Dictyostelium myosin II mutants that were previously phenotypically selected following random mutagenesis and shown to lie in the actin binding domain [Patterson, B., & Spudich, J. A. (1996) Genetics 143, 801-810]. We show that the conditional loss of myosin-dependent activity in vivo, which results from the mutations E531Q, P536R, and R562L, is likely due to the loss of important contacts with actin. Purified wild-type and mutant myosin subfragments 1 (S1), expressed in Dictyostelium, are alike in binding to actin and releasing it in an ATP-dependent manner. Furthermore, the rates of ATP hydrolysis without actin are similar for the mutant and wild-type S1s. Thus, the mutations in the actin binding site have little effect on ATP binding or product release in the absence of actin. All three mutants, however, have impaired actin activated ATPase activity, with apparent second-order rate constants for actin interactions that are 4-25-fold smaller than that of wild-type S1 at 30 degrees C. The mutations also cause defects in the ability to move actin, as measured by in vitro motility assays of full-length myosins. On the basis of motility of a mixture of wild-type and mutant myosins, there appears to be at least two classes of mutations, with the primary defect in either a weak or a strong actin binding state. In summary, the activities in vitro of myosins with mutations in the actin binding site suggest losses of important contacts with actin. PMID- 9214291 TI - Isolation and chemistry of the mixed anhydride intermediate in the reaction catalyzed by dethiobiotin synthetase. AB - Dethiobiotin synthetase (DTBS) catalyzes the formation of the cyclic urea, dethiobiotin (DTB), from (7R,8S)-diaminononanoic acid (DAPA), CO2, and ATP; the other products of the reaction are ADP and Pi. The first intermediate in the reaction sequence is the 7-carbamate of DAPA [Huang, W., et al. (1995) Biochemistry 34, 10985-10995; Gibson, K. J., et al. (1995) Biochemistry 34, 10976 10984; Alexeev, D., et al. (1995) Structure 3, 1207-1215]. The existence of the second postulated intermediate, a mixed carbamic-phosphoric anhydride formed when the carbamate is phosphorylated by ATP, is consistent with the cleavage of the gamma-phosphoryl group of ATP seen in DTBS reaction mixtures [Baxter, R. L., & Baxter, H. C. (1994) J. Chem. Soc., Chem. Commun., 759-760]. Two more direct lines of evidence for the mixed anhydride intermediate have now been obtained. First, a DTBS reaction mixture containing [18O]CO2 produced 18O-enriched DTB and Pi, as the existence of such an intermediate would require. Second, a moderately stable intermediate that could be labeled with either 14CO2, [gamma-33P]ATP, [9 3H]DAPA, or [1,7-14C]DAPA was trapped by quenching DTBS reactions at pH 4 and isolated by thin-layer chromatography. As expected for the proposed mixed anhydride, this species underwent acid hydrolysis to DAPA, CO2, and Pi; under basic conditions, the intermediate cyclized, yielding DTB and Pi. When returned to fresh enzyme at pH 7.5, the intermediate underwent cyclization at a rate comparable to that of normal turnover. PMID- 9214292 TI - Kinetic mechanism of human inosine 5'-monophosphate dehydrogenase type II: random addition of substrates and ordered release of products. AB - IMP dehydrogenase (IMPDH) catalyzes the oxidation of IMP to XMP with the concomitant reduction of NAD to NADH. This reaction is the rate-limiting step of guanine nucleotide biosynthesis. IMPDH is a target of immunosuppressive, antiviral, anticancer, and antiparasitic chemotherapy. We have determined a minimal kinetic mechanism for human IMPDH type II using NAD analogs, isotope effects, hydride exchange, and presteady state kinetics. The values of kcat for the NAD analogs are similar despite a great variation in the structure and reactivity of the compounds. This observation suggests that a common step is rate limiting, i.e., either hydrolysis of the E-XMP* intermediate or release of XMP. No Vm isotope effect is observed when 2-2H-IMP is the substrate, which indicates that hydride transfer is fast. This conclusion is confirmed by the observation of a burst of NADH production under presteady state conditions. These observations further suggest that either E-XMP* hydrolysis or XMP release is rate-limiting. V/Km deuterium isotope effects are observed for both substrates (1.9 for IMP and 2.5 for NAD), which indicates that substrate association is random. This result contradicts previous conclusions based on product inhibition studies. No NADH consumption is observed in the presence of XMP and IMPDH, which indicates that the overall reaction is irreversible. NADH consumption is observed in the presence of thio-NAD, IMP, and enzyme. These observations indicate that NADH traps the E-XMP* intermediate and demonstrates that hydride transfer is reversible. At infinite NADH, all of E-XMP* is trapped by NADH, as indicated by the equivalence of the rates of consumption of thio-NAD and NADH. Therefore the release of products is ordered, with NADH release preceding hydrolysis of E-XMP*. PMID- 9214293 TI - Mechanism of the Clostridium thermoaceticum pyruvate:ferredoxin oxidoreductase: evidence for the common catalytic intermediacy of the hydroxyethylthiamine pyropyrosphate radical. AB - The cofactor content and mechanism of pyruvate:ferredoxin oxidoreductase (PFOR) are controversial. By using rapid freeze-quench EPR and stopped-flow spectroscopy, the elementary steps that constitute the first half-reaction of the Clostridiumthermoaceticum PFOR mechanism were elucidated. A hydroxyethyl-TPP (HE TPP) radical was identified and characterized as a transient intermediate, and for the first time, the kinetic competence of this substrate-derived radical was demonstrated. When the C. thermoaceticum PFOR was reacted with pyruvate and CoA, it had a lifetime of only approximately 100 ms. The results described here suggest that this radical intermediate is often not detected in studies of alpha ketoacid oxidoreductases because it rapidly decays. It is postulated here that the HE-TPP radical is an intermediate in the mechanism of all PFORs irrespective of the number of 4Fe-4S clusters and will be detected in all PFORs when rapid mixing methods are used. The C. thermoaceticum PFOR was shown to contain two 4Fe 4S clusters, as concluded earlier [Wahl, R. C., & Orme-Johnson, W. H. (1987) J. Biol. Chem. 262, 10489-10496]. The first reductive half-reaction was shown to involve the following steps: (i) reaction with pyruvate with PFOR to form the hydroxyethylidene-TPP intermediate; (ii) one-electron transfer to reduce one of the two Fe4S4 clusters and yield the HE-TPP radical; and, (iii) reaction with CoA resulting in formation of acetyl-CoA, rapid decay of the HE-TPP radical intermediate, and reduction of the second Fe4S4 cluster. Thus, at the end of the first half-reaction, the two Fe4S4 clusters are fully reduced. The rate of the third step was found to depend on the CoA concentration (k = 35 per s at saturating concentrations of CoA); however, in its absence, this step was slower by approximately 4400-fold. PMID- 9214294 TI - Enterobactin biosynthesis in Escherichia coli: isochorismate lyase (EntB) is a bifunctional enzyme that is phosphopantetheinylated by EntD and then acylated by EntE using ATP and 2,3-dihydroxybenzoate. AB - In Escherichia coli, the siderophore molecule enterobactin is synthesized in response to iron deprivation by formation of an amide bond between 2,3 dihydroxybenzoate (2,3-DHB) and l-serine and formation of ester linkages between three such N-acylated serine residues. We show that EntB, previously described as the isochorismate lyase required for production of 2,3-DHB, is a bifunctional protein that also serves as an aryl carrier protein (ArCP) with a role in enterobactin assembly. EntB is phosphopantetheinylated near the C terminus in a reaction catalyzed by EntD with a kcat of 5 min-1 and a Km for apo-EntB of 6.5 microM. This holo-EntB is then acylated with 2,3-DHB in a reaction catalyzed by EntE, previously described as the 2,3-DHB-AMP ligase, with a kcat of 100 min-1 and a Km of <<1 microM for holo-EntB. The N-terminal 187 amino acids of EntB (isochorismate lyase domain) are not needed for reaction of EntB with either EntD or EntE as demonstrated by the equivalent catalytic efficiencies of the full length EntB (residues 1-285) and the C-terminal EntB ArCP domain (residues 188 285) as substrates for both EntD and EntE. PMID- 9214295 TI - Horseradish peroxidase oxidation of tyrosine-containing peptides and their subsequent polymerization: a kinetic study. AB - Tyrosine-containing model peptides were oxidized by horseradish peroxidase (HRP). This led to a peptide polymerization via condensation of the aromatic rings. Dimers, trimers, and tetramers (depending on the peptide length and on the position of the tyrosine in the sequence) were identified by electron spray mass spectroscopy. The second-order rate constants of the second step of the HRP reduction (CII --> E) was decreased by the presence of a positively charged amino group in the vicinity of the aromatic ring as determined by stopped flow measurements [k3 = 19 398 M-1 s-1 and k3 = 1016 M-1 s-1 for N-acetyltyrosine (NAT) and l-Tyr oxidations, respectively]. High-performance liquid chromatography was used to follow the kinetics of polymerization of some model peptides after their enzymatic oxidation. The first polymerization products exhibited a strong inhibitory effect toward further oxidation by HRP. This effect was not observed when using manganese-dependent peroxidase (MnP) which does not bind directly to the tyrosine residue but rather acts as a "distant catalyst". Saturation of the HRP was achieved with Pro-Gln-Gln-Pro-Tyr (kcat = 58 s-1, = 2.1 mM), NAT (kcat = 94 s-1, = 5.6 mM), and Gly-Tyr (kcat = 175 s-1, = 10.8 mM). Analysis of steady state kinetics of the reaction showed that the dimers formed initially behaved like competitive inhibitors. The value of the dissociation constant between HRP and dimers was 20 microM. A simplified model which accounts for these observations, including the formation of a Michaelis-Menten-like complex involving the donor and enzyme, is proposed and discussed. PMID- 9214296 TI - Phosphatidylinositol-specific phospholipase C cyclic phosphodiesterase activity depends on solvent polarity. AB - Large enhancements (maximum of 82-fold in terms of enzyme efficiency, Vmax/Km) of bacterial PI-PLC cyclic phosphodiesterase activity were observed in the presence of organic solvents miscible in water (dimethyl sulfoxide, dimethylformamide, and 2-propanol). In general, organic solvents lowered the Km for myo-inositol 1,2 cyclic phosphate (cIP) and increased Vmax substantially. This kinetic effect was similar to that obtained with phosphatidylcholine micelles and bilayers in an aqueous assay system for cyclic inositol phosphate hydrolysis [Zhou, C., et al. (1997) Biochemistry 36, 347-355]. Solvent properties were examined to determine which ones correlated with the activation of PI-PLC toward cIP in each solvent. Activation correlated best with the solvent polarity as measured by ET(30); no significant correlation was observed with solution surface tension, the bulk dielectric constant (epsilon), 1/epsilon (a measure of the strength of charge interactions), or the Hildebrand solubility parameter. The sigmoidal curve of the enzyme activity versus solvent polarity was consistent with the solvent promoting a transition in the enzyme from a low-activity to a high-activity form. Possible candidates for this change, including enzyme dimerization, helix B/loop stabilization, and dehydration of the active site, are discussed. PMID- 9214297 TI - Proteolytic mapping and substrate protection of the Escherichia coli melibiose permease. AB - The topology and substrate-induced conformational change(s) of the Na+ (Li+ or H+)-melibiose cotransporter (MelB) of Escherichia coli were investigated by limited protease digestion. To facilitate these analyses, MelB was epitope-tagged both at its carboxyl-terminus and at its amino-terminus. Limited digestion with different proteases indicates that the cytoplasmic loops connecting transmembrane domains 4-5, 6-7, and 10-11 together with the carboxyl-terminus of MelB are exposed in the cytoplasm. In contrast, periplasmic loops are highly resistant to all the proteases examined, including nonspecific proteases such as proteinase K and thermolysin. The effect of Na+ or Li+ and/or melibiose on the rate of protease digestion of the cytoplasmic loops was also analyzed. The rate of protease digestion of loop 4-5 is specifically reduced, by approximately 3-fold, by the presence of Na+ or Li+. These results suggest that loop 4-5 is near or part of the cation binding site. Moreover, the presence of both melibiose and either Na+ or Li+ further reduced the rate of protease digestion of this loop 4-5 by up to 9-fold, although no protection from protease digestion was observed when melibiose was added alone. The increase in resistance to proteases observed in the presence of the cation alone or the cation plus melibiose suggests that the interaction of the two cosubstrate with MelB results in change(s) of MelB conformation. PMID- 9214298 TI - Endothelial nitric oxide synthase: modulations of the distal heme site produced by progressive N-terminal deletions. AB - cDNAs coding for bovine endothelial nitric oxide synthase (eNOS) with N-terminal deletions of 52, 91, and 105 amino acids were constructed, and the proteins were expressed in Escherichia coli and purified by affinity chromatography. All three truncated proteins bind heme and exhibit the ferrous-CO absorption maximum at 444 nm characteristic of thiolate heme ligation. Deletion of the first 52 amino acids yields a fully active dimeric protein with the same spectroscopic properties as the wild-type. The myristoylation, palmitoylation, and polyproline domains of the enzyme located in the deleted region are therefore not required for full catalytic activity. The delta91 and delta105 proteins, which exhibit altered dimerization equilibria, retain 20 and 12%, respectively, of the maximal activity. Resonance Raman and UV-vis spectroscopy indicate that, in the absence of tetrahydrobiopterin (H4B) and l-Arg, the wild-type and delta52 proteins are predominantly five coordinate high spin, whereas the delta91 and delta105 proteins are six coordinate low spin. The delta91 and delta105 mutants bind H4B, as indicated by a concomitant decrease in the low-spin component of the UV-vis spectrum, but the binding of l-Arg is extremely slow ( approximately 15 min). Dithiothreitol readily coordinates as the sixth iron ligand in the delta91 and delta105 mutants but not in the delta52 or wild-type proteins. The dithiothreitol can be completely displaced by l-Arg but not by H4B. Resonance Raman comparison of wild-type eNOS and nNOS confirms that, in the absence of H4B and l-Arg, eNOS is primarily high spin whereas nNOS is predominantly six coordinate, low spin. The results indicate that Cys-101 is not critical for the binding of H4B and imply that some of the protein residues involved in dimer formation and in preservation of active site integrity are located, probably at the monomer monomer interface, in the N-terminal end of the protein. PMID- 9214299 TI - Globoside as a membrane receptor: a consideration of oligosaccharide communication with the hydrophobic domain. AB - Recognition of macromolecules by glycosphingolipids is closely correlated with the nature of the glycolipid carbohydrate; however, it is also thought to be secondarily modulated by the structure of the single fatty acid. In the present work, we sought insight into what physical effect a change in this fatty acid has on the extramembranous portion of globosides at liposomal surfaces mimicking systems for which modulated receptor function has been recorded in the past. Protons of the exocyclic hydroxymethyl group on the terminal Gal residue of globotriaosylceramide (Gb3) were replaced with deuterium. In this location, the nonperturbing probe nuclei sampled cumulative conformational and orientational characteristics of the oligosaccharide chain at a sugar residue that is critical in specific binding of verotoxins. Deuterated Gb3 having 18:1 fatty acid was compared to the same species having 22:1 fatty acid, at 6.3 mol % in unsonicated bilayers of dipalmitoylphosphatidylcholine/cholesterol. Both produced narrow, apparently axially asymmetric 2H NMR spectra over a wide temperature range. Motional properties of the terminal sugar were measurably influenced by the fluidity of the host matrix; however, evidence was not found for conformational or orientational variation in this sugar brought about by the fatty acid alteration. In related experiments, acetate protons on the terminal N-acetyl galactosamine (GalNAc) residue of globotetraosylceramide (Gb4) were substituted with deuterium, and the natural fatty acid was replaced with 18:0 or 24:0 species deuterated at C2. Once again, species with short vs long fatty acid were examined for evidence of headgroup differences. Spectra of Gb4 were compared at 10 mol % in unsonicated fluid bilayers of 1-palmitoyl-2-oleoylphosphatidylcholine, and at 5 mol % in membranes containing 33 mol% cholesterol. Spectral splittings reflecting cumulative effects on conformation and order at the terminal deuterated sugar remained unchanged between species having 18:0 vs 24:0 fatty acid in POPC/cholesterol. In a pure POPC host matrix, there was clear evidence of a motional difference between the two--the longer chain Gb4 demonstrating spectral asymmetry--but the spectral width was unchanged. Transverse relaxation times, T2, were measured. Our findings appear to help correlate the conclusions of a number of workers dealing with the molecular basis of crypticity. We suggest that changes in glycolipid receptor function based on ceramide fatty acid variation have a major origin in the fatty acid's ability to determine the thermodynamics of interaction with the host matrix, as reflected in such parameters as glycolipid motional properties, local membrane curvature, and likely glycolipid time-dependent lateral associations. The result at low concentrations of glycolipid may often be only a subtly altered collective surface epitope, best detected by a specific recognition event. PMID- 9214300 TI - Influence of iron-removal procedures on sequential electron transfer in photosynthetic bacterial reaction centers studied by transient EPR spectroscopy. AB - Electron spin polarized electron paramagentic resonance (ESP EPR) spectra were obtained with deuterated iron-removed photosynthetic bacterial reaction centers (RCs) to specifically investigate the effect of the rate of primary charge separation, metal-site occupancy, and H-subunit content on the observed P865+QA- charge-separated state. Fe-removed and Zn-substituted RCs from Rb. sphaeroides R 26 were prepared by refined procedures, and specific electron transfer rates (kQ) from the intermediate acceptor H- to the primary acceptor QA of (200 ps)-1 vs (3 6 ns)-1 were observed. Correlation of the transient EPR and optical results shows that the observed slow kQ rate in Fe-removed RCs is H-subunit-independent, and, in some cases, independent of Fe-site occupancy as Zn2+ substitution does not ensure retention of the native kQ. In addition, shifts in the optical spectrum of P865 and differences in the high-field region of the Q-band ESP spectrum for Fe removed RCs with slow kQ indicate possible structural changes near P865. The experimental X-band and Q-band spin-polarized EPR spectra for deuterated Fe removed RCs where kQ is at least 15-fold slower at room temperature than the (200 ps)-1 rate observed for native Fe-containing RCs have different relative amplitudes and small g-value shifts compared to the spectra of Zn-RCs which have a kQ unchanged from native RCs. These differences reflect the trends in polarization predicted from the sequential electron transfer polarization (SETP) model [Morris et al. (1995) J. Phys. Chem. 99, 3854-3866; Tang et al. (1996) Chem. Phys. Lett. 253, 293-298]. Thus, SETP modeling of these highly resolved ESP spectra obtained with well-characterized proteins will provide definitive information about any light-induced structural changes of P865, H, and QA that occur upon formation of the P865+QA- charge-separated state. PMID- 9214301 TI - Temperature dependence of the Qy resonance Raman spectra of bacteriochlorophylls, the primary electron donor, and bacteriopheophytins in the bacterial photosynthetic reaction center. AB - Qy-excited resonance Raman spectra of the accessory bacteriochlorophylls (B), the bacteriopheophytins (H), and the primary electron donor (P) in the bacterial photosynthetic reaction center (RC) of Rhodobacter sphaeroides have been obtained at 95 and 278 K. Frequency and intensity differences are observed in the low frequency region of the P vibrational spectrum when the sample is cooled from 278 to 95 K. The B and H spectra exhibit minimal changes of frequencies and relative intensities as a function of temperature. The mode patterns in the Raman spectra of B and H differ very little from Raman spectra of the chromophores in vitro. The Raman scattering cross sections of B and H are 6-7 times larger than those for analogous modes of P at 278 K. The cross sections of B and of H are 3-4 times larger at 95 K than at 278 K, while the cross sections of P are approximately constant with temperature. The temperature dependence of the Raman cross sections for B and H suggests that pure dephasing arising from coupling to low-frequency solvent/protein modes is important in the damping of their excited states. The weak Raman cross sections of the special pair suggest that the excited state of P is damped by very rapid (<<30 fs) electronic relaxation processes. These resonance Raman spectra provide information for developing multimode vibronic models of the excited-state structure and dynamics of the chromophores in the RC. PMID- 9214302 TI - Thermodynamics of binding of the distal calcium to manganese peroxidase. AB - We previously demonstrated that manganese peroxidase from Phanerochaete chrysosporiumwas susceptible to thermal inactivation due to release of the distal calcium, which maintained the distal heme environment of the enzyme [Sutherland, G. R. J., Zapanta, L. S., Tien, M., & Aust, S. D. (1997) Biochemistry 36, 3654 3662]. In this investigation the binding of calcium to the distal calcium binding site of manganese peroxidase was studied by optical absorption spectroscopy and isothermal titration calorimetry. The dissociation constant for the distal calcium binding site was 11 +/- 1 microM and the Hill coefficient was 1.1 +/- 0.1. The binding of calcium was accompanied by decreases in enthalpy and entropy that were large compared to those of other calcium binding proteins. The decreases were consistent with the large conformational changes proposed to occur in manganese peroxidase as a result of the binding and release of the distal calcium. Studies involving binding of the hydrophobic fluorescent probe, 4,4' dianilino-1,1'-binaphthyl-5,5'-disulfonic acid, dipotassium salt (bis-ANS), to manganese peroxidase indicated that the active, calcium-containing form of the enzyme had less exposed hydrophobic surface area, which would contribute to an increase in enthalpy and entropy upon calcium binding. Therefore, the negative changes in enthalpy and entropy associated with calcium binding were attributed to a large increase in the structural rigidity and compactness of the enzyme. The dissociation constant for calcium decreased and the rate of thermal inactivation decreased with decreasing pH. However, both the ability of calcium to prevent thermal inactivation of manganese peroxidase and the rate of calcium binding decreased as the pH decreased. Therefore it was proposed that, at lower pH, calcium binding to manganese peroxidase was more thermodynamically favorable, but the rate of calcium binding decreased because the flexibility of the calcium binding site, and in turn exposure of the ligands to the incoming ion, decreased. PMID- 9214303 TI - Reduction of thiocyanate, cyanate, and carbon disulfide by nitrogenase: kinetic characterization and EPR spectroscopic analysis. AB - Nitrogenase catalyzes the reduction of N2, protons, and a number of alternative substrates that contain C-C, C-N, N-N, and N-O double and triple bonds. Recently it has been shown that nitrogenase also reduces the C==S bond of COS and the C==O bond of CO2. The current work demonstrates that the COS analogs SCN-, CS2, and OCNH are novel substrates for nitrogenase and that the reduction of these substrates produces changes in the electron paramagnetic resonance (EPR) spectrum of nitrogenase, providing insight into the mechanism of substrate reduction by nitrogenase. CH4, HCN, H2S, and NH4+ were detected as products of the nitrogenase catalyzed reduction of SCN-. CS2 was reduced by nitrogenase to H2S, providing the first demonstration of CS2 reduction catalyzed by a purified enzyme. CO was detected as a product of KOCN reduction by nitrogenase. Interestingly, the Km for KOCN reduction to CO decreased at lower pH values, suggesting that OCNH rather than OCN- was the substrate for nitrogenase. Analysis of the EPR spectra of nitrogenase under turnover conditions in the presence of KOCN, CS2, or KSCN revealed new EPR signals. Signals with g-values corresponding to those reported for CO bound to the iron-molybdenum cofactor of nitrogenase were detected during turnover of nitrogenase in the presence of KOCN. During SCN- and CS2 reduction by nitrogenase, novel EPR inflections were observed that appear to report the interaction between nitrogenase and a bound substrate or a transient intermediate produced during the reduction of SCN- and CS2. PMID- 9214304 TI - Pressure-induced dissociation of yeast glyceraldehyde-3-phosphate dehydrogenase: heterogeneous kinetics and perturbations of subunit structure. AB - In studies of pressure-induced subunit dissociation of oligomeric proteins, the thermodynamic dissociation constant and the dissociation volume change are derived by assuming that high pressure itself does not significantly perturb the structure of both oligomer and isolated subunit. In this report, the intrinsic phosphorescence emission of Trp reveals that high-pressure dissociation of tetrameric yeast glyceraldehyde-3-phosphate dehydrogenase results in a dramatic shortening of the phosphorescence lifetime, from 300 to less than 2 ms, that is consistent with a profound loosening of the polypeptide structure about the phosphorescence probe. On pressure release, subunit reassociation occurs readily whereas recovery of the native phosphorescence properties is a very slow, thermally activated, process which goes hand in hand with the recovery of the catalytic activity. Further, the comparison between the kinetic traces that describe the degree of dissociation and the change in phosphorescence lifetime, at various applied pressures, has established the following: (1) that high pressure plays a direct role on the structural rearrangement, the extent of which increases with pressure; (2) that the conformational change in the monomer is concomitant with, or follows closely after, the break up of the tetramer, in any case long before an apparent tetramer-monomer equilibrium is established; (3) that native tetramers are highly heterogeneous with regard to their rate of dissociation. The influence of temperature, of protein concentration, of binding of NAD+, and of the addition of 2 M urea on the dissociation/phosphorescence kinetic profiles was also examined. The complications arising from these conformational changes for the derivation of the dissociation free energy change as well as their relevance for understanding the lack of concentration dependence of the degree of dissociation are discussed. PMID- 9214305 TI - Fluorescence spectral changes during the folding of intestinal fatty acid binding protein. AB - Although large changes in fluorescence intensity are observed during the folding and unfolding of many proteins, it has been difficult to associate these changes with specific structures or with the environmental changes which a particular tryptophan may undergo during these processes. The fluorescence spectral changes that occur during the folding and unfolding of rat intestinal fatty acid binding protein (IFABP) are described here. The intermediate observed during unfolding had spectral characteristics similar to those of unfolded protein, but with somewhat higher intensity. Stopped-flow circular dichroism measurements during unfolding showed that little if any secondary structure was associated with this intermediate. During refolding, the initial fluorescence spectrum was not that of native or unfolded IFABP, suggesting that some structure with intermediate fluorescent properties had formed during the deadtime of mixing. The shape and intensity of this initial spectrum were dependent on the final urea concentration, becoming more native-like at lower final concentrations of denaturant. A simple model for refolding suggests that a portion of the protein molecules obtain native structure and fluorescent characteristics during the deadtime of mixing, and that the remaining protein molecules have spectral characteristics similar to those of the intermediate observed during unfolding. Lower final concentrations of denaturant cause a larger proportion of molecules to follow the rapid refolding pathway. Knowledge of the fluorescence spectral characteristics of the intermediates formed during the folding and unfolding of any protein will improve our understanding of the nature of these structures. PMID- 9214306 TI - Folding of tryptophan mutants of barstar: evidence for an initial hydrophobic collapse on the folding pathway. AB - The contributions of the three tryptophan residues of barstar to the spectroscopic properties, stability, and folding of the protein have been studied by mutating two of the tryptophans, Trp38 and Trp44, individually as well as together, to phenylalanines, Phe. The three mutant proteins studied are shown to be similar to wt barstar in structure by activity measurements as well as by spectroscopic characterization. Fluorescence energy transfer between the tryptophans as well as quenching by their local structural environments complicates the analysis of the contributions of the individual tryptophans to the fluorescence of the wt protein, but it is demonstrated that Trp53, which is completely buried within the hydrophobic core, makes the dominant contribution to the fluorescence, while the fluorescence of Trp38 is largely quenched in the fully folded protein. GdnHCl- as well as temperature-induced equilibrium unfolding studies, using three different structural probes, indicate that W38FW44F, where both Trp38 and Trp44 have been removed, follows a two-state unfolding transition and is less stable than the wt barstar. The fluorescence monitored folding and unfolding kinetics of W38FW44F have been studied in detail. W38FW44F folds 2-fold faster and unfolds 3-fold faster than wt barstar. A large fraction of the total fluorescence change that occurs during folding occurs in a burst phase within 4 ms after commencement of folding. A similar burst phase change in fluorescence, although to a smaller extent, is shown to occur during the folding of wt barstar. The results suggest that a very early folding intermediate accumulates within 4 ms of folding, and that this kinetic intermediate is sufficiently compact that Trp53, which is completely sequestered from solvent in the fully folded protein, is also significantly sequestered from solvent in this intermediate. PMID- 9214307 TI - 13C NMR analysis of the cysteine-sulfenic acid redox center of enterococcal NADH peroxidase. AB - In order to characterize the native Cys42-sulfenic acid redox center of the flavoprotein NADH peroxidase by NMR, an expression protocol has been developed which yields the [3-13C]Cys42-labeled protein in 100 mg quantities. Difference spectra of the labeled minus unlabeled oxidized enzyme (E) give a peak at 41.3 ppm (relative to dioxane) which represents the Cys42-sulfenic acid. Reduction of labeled E with 1 equiv of NADH gives the air-stable two-electron reduced (EH2) species, and oxidized minus reduced difference spectra give maxima and minima at 41.3 and 30.8 ppm, respectively, corresponding to the Cys42-sulfenic acid and thiolate species. Peroxide inactivation of E, which has previously been attributed to oxidation of the Cys42-sulfenic acid to the Cys42-sulfinic and/or sulfonic acid states, gives rise to a new maximum in the difference spectrum of Einactive minus E at 57.0 ppm. A similar expression protocol was used to obtain the [ring-2-13C]His-labeled peroxidase HHAA mutant (His10His23Ala87Ala258); the spectral change over the pH range 5.8-7. 8 is attributed to deprotonation of the surface-exposed His23. Furthermore, replacement of Arg303, which is hydrogen bonded to His10, has no effect on the 13C spectrum. These results provide direct evidence in support of the peroxidase Cys42-sulfenic acid/thiol redox cycle and add significantly to our structure-based understanding of protein-sulfenic acid stabilization and function. PMID- 9214308 TI - Conformation of the principal neutralizing determinant of human immunodeficiency virus type 1 in complex with an anti-gp120 virus neutralizing antibody studied by two-dimensional nuclear magnetic resonance difference spectroscopy. AB - The principal neutralizing determinant (PND) of human immunodeficiency virus type 1 (HIV-1) is located in the third hypervariable region (V3) of the virus envelope glycoprotein gp120. The conformation of a V3 peptide of HIV-1IIIB bound to the Fab fragment of an anti-gp120 HIV neutralizing antibody, 0.5beta, was studied by 1H NMR spectroscopy. This 18-residue peptide represents the epitope recognized by 0.5beta and encompasses most of the PND. The slow off-rate of the peptide prevents the observation of peptide/Fab interactions as well as intramolecular interactions within the bound peptide by transferred nuclear Overhauser enhancement (TRNOE). To detect and assign interactions within the bound peptide in the 52 kDa complex, NOESY difference spectra were measured using three strategies: (a) deuteration of peptide residues, (b) Arg two head right arrow Lys replacements, and (c) truncation of the peptide antigen. Each difference spectrum was calculated between NOESY spectra measured for two Fab complexes in which the bound peptides differed in their deuteration or in their sequence. The difference spectra revealed numerous interactions between the N-terminus of the epitope (Arg 4, Lys-5, Ser-6, Ile-7, and Ile-9) and its C-terminus (Phe-17, Val-18, Thr-19, and Ile-20). The assigned NOE interactions within the bound peptide were translated into distance restraints that were used to calculate the conformation of the bound peptide by the hybrid distance geometry/simulated annealing method. A total of 39 long-range (residues i - j >> 4), 14 short-range, and 69 intraresidue NOE interactions within the bound peptide have been assigned. Twelve structures without NOE constraint violations were obtained, having a 1.6 A rms deviation for the backbone atoms. The peptide forms a 10-residue loop, while the two segments flanking this loop, KSI and VTI, interact extensively with each other and possibly form antiparallel beta-strands. This loop conformation could be observed due to the unusual large size (17 residues) of the antigenic determinant recognized by 0.5beta. PMID- 9214309 TI - Kinetics of oxidized cytosine repair by endonuclease III of Escherichia coli. AB - Endonuclease III of Escherichia coli excises a broad range of oxidized, hydrated and ring-fragmented pyrimidines from DNA. The kinetic parameters were compared for repair of three potentially mutagenic oxidized cytosine lesions: 5,6 dihydroxy-5, 6-dihydro-2'-deoxyuridine (uracil glycol or Ug), 5-hydroxy-2' deoxycytidine (5-ohC), and 5-hydroxy-2'-deoxyuridine (5-ohU). Site-specifically modified 40-mer oligonucleotides containing each of the three lesions in the same sequence context were synthesized chemically or by a combination of chemical and enzymatic methods. Appropriately protected phosphoramidites of 5-ohC and 5-ohU were synthesized and incorporated into oligonucleotides by standard solid-phase synthetic methods. The lability of Ug made it necessary to use an alternative approach to prepare the analogous 40-mers containing Ug. An uracil containing pentamer oligonucleotide was oxidized with OsO4 to generate the corresponding Ug containing product, which was then ligated into an oligonucleotide scaffold to generate 40 base pair duplexes. Using 32P-labeled substrates and a gel electrophoresis based assay, the values of Km and Vmax for excision of 5-ohC, 5 ohU, and Ug were determined. In this experimental system, the order of repair efficiency is Ug >> 5-ohC >> 5-ohU based on ratios of Vmax/Km. Modest effects were observed when the base paired opposite the lesion was changed from G to A. PMID- 9214310 TI - Selenocysteine tRNA[Ser]Sec levels and selenium-dependent glutathione peroxidase activity in mouse embryonic stem cells heterozygous for a targeted mutation in the tRNA[Ser]Sec gene. AB - To investigate the effect of a reduced level of selenocysteine (Sec) tRNA[Ser]Sec in selenoprotein biosynthesis, two mouse embryonic stem (ES) cell lines heterozygous for the corresponding gene were generated by homologous recombination of the host genome with targeting vectors encoding a deleted or a disrupted tRNA[Ser]Sec gene. The presence of a single functional gene in ES cells afforded us an opportunity to determine directly in the cell line the effect of reduced gene dosage on (1) the levels of the Sec tRNA[Ser]Sec population, (2) the distributions of the isoacceptors within the Sec tRNA population, and (3) selenoprotein biosynthesis. We therefore determined the amounts and distributions of the two major tRNA[Ser]Sec isoacceptors, designated mcm5U and mcm5Um, within the Sec tRNA population and determined the activity of the anti-oxidant, selenium containing glutathione peroxidase (GPx) in the heterozygotes and in wild type cells grown in media with and without added selenium. The level of the Sec tRNA[Ser]Sec population in the heterozygotes was approximately 60% of that of wild type cells grown in media under normal conditions, while the ratio of the mcmU isoacceptor in wild type vs mutant cells was approximately 2:1 and of the mcmUm isoacceptor approximately 1:1. In the presence of media supplemented with selenium, the Sec tRNA[Ser]Sec population increased about 20% in wild type cells and virtually not all in heterozygous cells, and the level of the Sec tRNA[Ser]Sec population was, therefore, approximately 50% of that of wild type cells. GPx activity was indistinguishable among these cell lines in either selenium-supplemented or unsupplemented media, indicating that the resultant changes in tRNA[Ser]Sec levels did not have a measurable effect on GPx biosynthesis. PMID- 9214311 TI - DNA strand scission by polycyclic aromatic hydrocarbon o-quinones: role of reactive oxygen species, Cu(II)/Cu(I) redox cycling, and o-semiquinone anion radicals,. AB - In previous studies, benzo[a]pyrene-7,8-dione (BPQ), a polycyclic aromatic hydrocarbon (PAH) o-quinone, was found to be 200-fold more potent as a nuclease than (+/-)-anti-7,8-dihydroxy-9,10-epoxy-7,8,9, 10-tetrahydrobenzo[a]pyrene, a suspect human carcinogen. The mechanism of strand scission mediated by naphthalene-1,2-dione (NPQ) and BPQ was further characterized using either phiX174 DNA or poly(dG).poly(dC) as the target DNA. Strand scission was extensive, dependent on the concentration of o-quinone (0-10 microM), and required the presence of NADPH (1 mM) and CuCl2 (10 microM). The production of reactive species, i.e., superoxide anion radical, o-semiquinone anion (SQ) radical, hydrogen peroxide (H2O2), hydroxyl radical (OH.), and Cu(I), was measured in the incubation mixtures. The formation of SQ radicals was measured by EPR spectroscopy under anaerobic conditions in the presence of NADPH. A Cu(II)/Cu(I) redox cycle was found to be critical for DNA cleavage. No strand scission occurred in the absence of Cu(II) or when Cu(I) was substituted, yet Cu(I) was required for OH* production. Both DNA strand scisson and OH. formation were decreased to an equal extent, albeit not completely, by the inclusion of OH. scavengers (mannitol, soduim benzoate, and formic acid) or Cu(I) chelators (bathocuproine and neocuproine). In contrast, although the SQ radical signals of NPQ and BPQ were quenched by DNA, no strand scission was observed. When calf thymus DNA was treated with PAH o-quinones, malondialdehyde (MDA) was released by acid hydrolysis. The formation of MDA was inhibited by OH. scavengers suggesting that OH* cleaved the 2'-deoxyribose moiety in the DNA to produce base propenals. These studies indicate that for PAH o-quinones to act as nucleases, NADPH, Cu(II), Cu(I), H2O2, and OH*, were necessary and that the primary species responsible for DNA fragmentation was OH., generated by a Cu(I)-catalyzed Fenton reaction. The genotoxicity of PAH o-quinones may play a role in the carcinogenicity and mutagenicity of the parent hydrocarbons. PMID- 9214312 TI - Structure/calcium affinity relationships of site III of calmodulin: testing the acid pair hypothesis using calmodulin mutants. AB - Calmodulin mutants in which the calcium binding affinity of site IV was greatly reduced by a D133E mutation were prepared using site-specific, cassette-mediated mutagenesis as a multisite calcium binding protein model to examine structure/calcium affinity relationships in site III of calmodulin. Tryptophan was introduced in position 92 of the calmodulin mutants as a fluorescent label to monitor the calcium-induced structural changes in the C-terminal domain of calmodulin. The five calmodulin mutants, 3xCaM, 3zCaM, 4xCaM, 4zCaM, and 4xzCaM, were designed so that there were three or four acidic amino acid residues in chelating positions of site III with acid pairs on either the X and/or Z coordinating axes. The calcium dissociation constant of site III, KIII, of the five calmodulin mutants changes in a descending order from 3xCaM (237 microM), 3zCaM (140 microM), 4xCaM (5.8 microM), 4zCaM (3 microM), to 4xzCaM (2 microM), and these KIII values are significantly lower than that of F92W/D133E calmodulin (335 microM) in which three acidic residues with no acid pairs were present in site III [Wu, X., & Reid, R. E. (1997) Biochemistry 36, 3608-3616]. These results indicate that the calcium affinity of site III increases when the number of the acidic chelating residues increases from three to four, when the number of acid pairs increases from zero to one and further to two, and when the location of the acid pair is changed from the X axis to the Z axis. This study provides the first evidence that the acid pair hypothesis which correlates the nature of the chelating residues with the calcium affinity of the hlh motif is applicable to a multisite calcium binding protein model. The Hill coefficients indicate that reversal of the sequence of filling of the calcium binding sites in the C terminal domain from IV --> III to III --> IV also changes the site cooperativity from positive to negative. The cooperativity returns to positive when the proteins are titrated in the presence of a calmodulin-binding peptide. Data from the present study also demonstrate that calmodulin mutants with a decreased calcium affinity have a reduced efficiency in phosphodiesterase regulation at low calcium concentrations (50 microM). However, high calcium concentrations (15 mM) restore the phosphodiesterase regulatory activity of the calmodulin mutants to a level obtained with F92W calmodulin, indicating that the mutations alter calcium regulation of calmodulin-mediated phosphodiesterase activity without affecting the interaction between calmodulin and the enzyme. PMID- 9214313 TI - The anionic phospholipid-mediated membrane interaction of the anti-cancer drug doxorubicin is enhanced by phosphatidylethanolamine compared to other zwitterionic phospholipids. AB - The interaction of doxorubicin and lipids has been studied using large unilamellar vesicles (LUVET) composed of mixtures of anionic phospholipids and various zwitterionic phospholipids. Dilution of anionic lipids with zwitterionic lipids leads to decreased membrane association of the drug because electrostatic forces are very important in doxorubicin-membrane interaction. However, binding of doxorubicin to LUVET composed of anionic phospholipids combined with phosphatidylethanolamine (PE) is much higher than binding to LUVET made of anionic lipids plus a range of other zwitterionic lipids such as phosphatidylcholine (PC) and the N-methylethanolamine and N, N dimethylethanolamine derivatives of PE. This preferential interaction is observed with all negatively charged phospholipids tested and is, in the case of phosphatidylserine (PS), confirmed in monolayer experiments. The increase in surface area observed in a monolayer composed of PS and PE (1/3) was 3 times higher than in a monolayer of PS/PC (1/3). The preferential interaction appears not to be due to the ability of PE to adopt inverted nonbilayer structures, but probably involves a combination of the ability of PE to form additional hydrogen bonds and of the intrinsic curvature of a bilayer containing PE because of its small headgroup. Implications of our finding for the in vivo membrane interaction and transport of the drug will be discussed. PMID- 9214315 TI - Acid-base balance alterations in laparoscopic cholecystectomy. AB - BACKGROUND: The purpose of this study is to determine alterations of acid-base balance originated by pneumoperitoneum with CO2. Influence of other factors such as anesthetic technique, duration of procedure, and volume of CO2 insufflated has also been analyzed. METHODS: Some 132 patients were divided in three groups according to anesthetic technique used. Arterial blood gases were determined before pneumoperitoneum, at 20 min after it, and every 30 min, until procedure's end, and in postoperative period up to a total of four samples. RESULTS: Pneumoperitoneum originated a fall of pH (p << 0. 001), ion bicarbonate (p << 0.001), and base excess (p << 0.001) and an elevation of PaCO2 (p << 0.001). No correlation was found between these changes and duration of pneumoperitoneum or amount of CO2 insufflated. Changes were fundamentally of a metabolic type. There were no statistically significant differences among anesthetic techniques. CONCLUSIONS: In conclusion, pneumoperitoneum with CO2 originates alterations of the acid-base balance, mostly of a metabolic type. This could mean that besides CO2 absorption, there is a tissular hypoperfusion due to the increase of abdominal pressure. PMID- 9214314 TI - Lobectomy with extended lymph node dissection by video-assisted thoracic surgery for lung cancer. AB - BACKGROUND: Between September 1992 and September 1996, we performed 88 VATS (video-assisted thoracic surgery) lobectomies and two VATS pneumonectomies. METHODS: The indications for surgery were 68 cases of lung cancer, nine cases of bronchiectasis, six cases of tuberculosis, and seven cases of benign lesions. Of the 68 cases of lung cancer, 36 were treated by VATS lobectomy with extended lymph node dissection for clinical stage I lung cancer, making full use of recently developed devices for thoracoscopic surgery, such as roticulating endoscissors, miniretractors, endoclips, and harmonic scalpels. RESULTS: Twenty four lymph nodes were resected on average (range, 10 to 51) by VATS. This number was comparable to lymph nodes resected in open thoracotomy during the same period. Among the 36 patients who underwent extended lymph node dissection, 20 showed no lymph node metastasis postoperatively (stage I), while 16 had N1 or N2 cancer. All patients with stage I cancer have survived 4 to 36 months (median: 17 months) with no signs of recurrence. CONCLUSIONS: This survival of stage I lung cancer after VATS is comparable to that of open thoracotomy. We thus believe that VATS lobectomy with extended lymph node dissection can be an alternative to standard posterolateral thoracotomy for stage I lung cancer. PMID- 9214316 TI - The effects of sterile or infected bile and dropped gallstones in abdominal adhesions and abscess formation. AB - BACKGROUND: The effects of gallstones and sterile or infected bile on postoperative adhesions and abscess formation were investigated in Sprague Dawley rats. METHODS: The first three groups were injected intraperitoneally with serum saline, sterile bile, or infected bile, respectively. Laparotomy was adjusted to the next seven groups. Serum saline, sterile bile, and infected bile were injected in the fourth through sixth groups intraperitoneally, respectively. Gallstones were placed intraabdominally to the seventh through ninth groups. The injections of sterile bile in group 7 and of infected bile in group 8 were added to laparotomies. Only laparotomy was carried out in group 10. RESULTS: Abscess formations were seen only in infected-bile-injected groups. Significant adhesion formations were seen in infected-bile groups. Sterile bile and/or gallstones had no significant effect in abscess or adhesion formation. CONCLUSIONS: Results suggest that noninfected gallstones and sterile bile, even in combination, do not increase postoperative intraabdominal complications in rats. PMID- 9214317 TI - Common bile duct repair with titanium staples. Comparison with suture closure. AB - BACKGROUND: Vascular Closure Staple (VCS) clips made of titanium were initially developed for microvascular anastomoses with little experience of their use in larger tubular structures. This study compares VCS clips and sutures in the closure of supraduodenal choledochotomy. METHODS: In nine pigs, two longitudinal incisions of the common bile duct (CBD) were randomly assigned to closure with 4 0 interrupted polyglactin sutures or VCS clips. RESULTS: Clip closure was significantly faster (116 +/- 28 vs 581 +/- 88 s). All nine CBDs were patent and without signs of calculus formation after 3 months. Clip closure resulted in slightly less narrowing of the duct lumen and thinner scar at the repair site. At histological examination, all 18 incisions had healed without signs of fistula formation or marked fibrosis. CONCLUSIONS: Choledochotomy closure with VCS clips results in as good or better wound healing than suture closure, with a comparable degree of narrowing. The time required for clip closure is only about one-fifth that of suture closure. PMID- 9214318 TI - Laparoscopic management of acute biliary pancreatitis. AB - BACKGROUND: The appropriate management of acute biliary pancreatitis has evolved considerably over the past decades. The advent of laparoscopic surgery made it necessary to reevaluate the traditional algorithms. METHODS: This study assesses the outcome of 40 patients laparoscopically treated for gallstone pancreatitis. The severity of pancreatitis was scored by clinical and biochemical evaluation and CT findings. Laparoscopic cholecystectomy was performed during the same admission in all cases. In no case was a preoperative endoscopic retrograde cholangiopancreatography (ERCP) performed. In 32 patients (80%) with mild acute pancreatitis interval cholecystectomy was less than 1 week (group A) and in eight patients (20%) with severe disease it was more than 7 days (group B). All patients underwent intraoperative cholangiography. RESULTS: The rate of common bile duct (CBD) stones was 5% (two cases), both occurring in the group A. There was one perioperative death (2.5%) in group B and one late CBD injury (2.5%) in group A, not requiring surgery. Complication rate was significantly higher in group B (50%) than in group A (9.4%). CONCLUSIONS: We consider that treatment of mild-to-moderate acute biliary pancreatitis can be satisfactorily accomplished by laparoscopy with routine intraoperative cholangiography and laparoscopic treatment of bile duct stones, showing no mortality and low morbidity rate. Laparoscopic treatment of patients with severe acute pancreatitis deserves further investigation. PMID- 9214319 TI - A stratified intraoperative surgical strategy is mandatory during laparoscopic common bile duct exploration for common bile duct stones. Lessons and limits from an initial experience of 92 patients. AB - BACKGROUND: Open exploration and endoscopic sphincterotomy (ES) remain the preferred treatment of common bile duct stones (CBDS). The recent spread of laparoscopy has worsened the dilemna of choosing between surgical and endoscopic treatment of CBDS. The aim of this study was to critically evaluate the results of our preliminary experience with laparoscopic common bile duct exploration (CBDE) for CBDS. METHODS: Ninety-two consecutive patients were prospectively submitted to laparoscopic CBDE. Surgical strategy included an initial transcystic approach or laparoscopic choledochotomy. Failure of stone clearance was managed by conversion to open CBDE or by postoperative ES. Electrohydraulic lithotripsy and papillary balloon dilatation were selectively used. Stone clearance was assessed by choledochoscopy and control cholangiography. RESULTS: The overall laparoscopic stone clearance in this series was 84% (transcystic route 63% and choledochotomy 93%). Conversion to laparotomy was mandatory in 12% of the patients because of incomplete stone clearance and in 5% because of intraoperative complications. Postoperative ES was required in 4% of the patients, giving an overall surgical success rate of 96%. When indicated (small and limited number of stones located below the cysticocholedochal junction, with a dilated and patent cystic duct) the transcystic route had the lower success rate, the higher complication rate, and the shorter operative time and postoperative hospital stay. When indicated (accessible and dilated common bile duct over 7 mm), laparoscopic choledochotomy had the higher success rate, the lower complication rate, the longer operative time, and the longer postoperative hospital stay, which is related to associated external biliary drainage. The hospital mortality included two high-risk patients (2%) and the complications rate was 15%. CONCLUSIONS: Laparoscopic CBDE is safe in selected patients. A stratified intraoperative surgical strategy is mandatory in deciding between a transcystic route and choledochotomy with specific indications for each approach. When feasible, laparoscopic choledochotomy is more efficient and safe than the transcystic route, but it is associated with a longer postoperative hospital stay, which is due to external biliary drainage. PMID- 9214320 TI - Laparoscopically guided bipolar radiofrequency ablation of areas of porcine liver. AB - BACKGROUND: Bipolar radiofrequency ablation (BRFA) is a promising technique with which to treat unresectable primary and metastatic liver tumors. Its effects on normal liver tissue and postoperative liver function, however, are unknown. We performed this study to determine (1) the feasibility of using laparoscopic ultrasound to guide placement of BRFA needle electrodes in the liver and (2) the histopathologic, hepatic biochemical, and systemic hemodynamic responses to BRFA. METHODS: Two BRFA lesions were created in the liver of adult domestic pigs to ablate 8-10% of the normal liver volume. Laparoscopic ultrasound was used to guide creation of one peripheral liver lesion and one central liver lesion (with a major hepatic or portal venous vein branch in the center of the BRFA lesions) in each animal. BRFA of liver tissue was performed by passing 12 W of RF power for 16 min across two 16-gauge active-needle electrodes placed 3 cm apart. RESULTS: All animals survived the procedure without significant hemodynamic alterations during or after BRFA. All animals had a transient elevation in serum transaminase levels that returned to normal within 1 week of the BRFA of liver tissue. Gross and microscopic histopathology of the BRFA lesions revealed 2.0-2.5 cm zones of complete coagulative necrosis around and between the BRFA needle tracks without destruction of major blood vessel walls. CONCLUSIONS: This study demonstrates (1) that laparoscopic ultrasound can be used to guide placement of BRFA needles in the liver and (2) that BRFA produces focal destruction of liver without significant systemic hemodynamic responses or alterations in liver function. Further studies of this technique to ablate malignant liver tumors are ongoing. PMID- 9214321 TI - Laparoscopic treatment of nonparasitic cysts of the liver with omental transposition flap. AB - BACKGROUND: Between 1991 and November 1994, 18 patients with large, solitary, nonparasitic liver cysts underwent laparoscopic deroofing; the last 13 of them also received an omental transposition flap in addition. METHODS: Using three to four trocars, the cystic contents were first aspirated, and the cyst derooted widely using diathermia. An omental transposition flap was fashioned and stapled into the cyst cavity itself. RESULTS: Postoperative complications included one case of pulmonary atelectasis. Another patient developed a subhepatic bile collection which was aspirated percutaneously. On average, patients were discharged on the 4th (2-14) postoperative day. Follow-up was performed with abdominal ultrasound for 2-43 months (mean 19 months). There were two early cyst recurrences, both in cases without an omental transposition flap (overall recurrence rate, 11%; in patients with omental flap, 0). CONCLUSIONS: Deroofing in combination with an omental transposition flap is a safe and effective therapy for symptomatic solitary liver cysts and can be performed using minimal-access surgical techniques. PMID- 9214322 TI - Abdominal fat tissue necrosis as a cause of acute abdominal pain. Laparoscopic diagnosis and therapy. AB - BACKGROUND: Infarctions of the greater omentum and of the epiploic appendages are rare etiologies of acute abdominal pain. The aims of the study were to determine the incidence of abdominal fat tissue necroses and to discuss the clinical features and the role of laparoscopy in the treatment of these conditions. METHODS: A retrospective study in 563 consecutive patients with acute abdominal pain was performed. In all patients diagnostic laparoscopy was indicated. RESULTS: The incidence of abdominal fat tissue necroses in 563 patients with acute abdominal pain was 1.1%. Six patients had either infarctions of the omentum or of the epiploic appendages. Pain was the predominant clinical symptom and the preoperative diagnosis depended upon the location of the omental or epiploic necroses. Diagnosis and treatment were performed laparoscopically without morbidity. CONCLUSION: The incidence of abdominal fat tissue necroses in our patients was increased compared to the prelaparoscopic period. Omental and epiploic necroses are significant in the differential diagnosis of appendicitis, acute cholecystitis, and diverticulitis. PMID- 9214323 TI - Percutaneous endoscopic gastrostomy (PEG). 8 years of clinical experience in 232 patients. AB - BACKGROUND: Percutaneous endoscopic gastrostomy (PEG) is now a standard method for providing long-term enteral nutrition in patients who are unable to swallow. The aim of our study was to document clinical data that would allow prediction of a possible complicated clinical course. METHODS: The study was carried out retrospectively. Clinical data of patients having received a PEG tube by a single endoscopic technique were analyzed. RESULTS: Some 5. 17% of 232 patients showed complications requiring surgery including a mortality rate of 0.43%. Patients with complications had a significantly lower body mass index and there was a significantly higher complication rate in patients having obstructive malignancies compared with benign diseases. CONCLUSIONS: Low body mass index and advanced malignancies are predictors for complications after PEG application. Early installation should help prevent further nutritional deterioration and the related risk of complications. PMID- 9214324 TI - Laparoscopic colectomy. AB - BACKGROUND: Laparoscopic colectomy has developed with the explosion of technology that has followed laparoscopic cholecystectomy. Accumulation of skills in general laparoscopic surgery has made complex surgery, such as colectomy, feasible. METHODS: Three hundred fifty-nine laparoscopic cases were prospectively studied. Data has been kept on benign and malignant cases, operative results, hospital stay, and morbidity. Special care has been taken to follow malignant cases, looking for recurrence of disease. RESULTS: There were 359 cases (206 females, 153 male) average age 58.8 years (18-94), and 149 patients had malignancy. All types of resections were performed, including 151 anterior resections, 66 right hemicolectomies (RHC), 36 total colectomies, and 22 rectopexies. Operating times fell with experience-the last 20 cases of anterior resection took 150 min (110 240) and of RHC took 130 min (65-210). Twenty-six (7%) cases were converted to open surgery. Hospital stays for anterior resection lasted 5-7 days (2-33); in the last 20 cases the average stay was 4 days. Morbidity included seven leaks (2.7%), four strictures (1.2%), 12 wound infections (3.3%), and nine ileus (2.5%). There were six deaths within 30 days-sepsis, myocardial infarction, aspiration pneumonia, and disseminated liver metastases. One hundred forty-nine cancer cases have had ten recurrences: one pelvic recurrence, six liver metastases, two para-aortic nodal, and one case of disseminated disease. Average time of recurrence was 33 months (15-46 months). CONCLUSIONS: Laparoscopy in the hands of experienced laparoscopic surgeons is a safe, efficient procedure. All types of procedures are possible. Early results in 149 malignancies are encouraging and recurrence rates are low. Prospective studies, now that skills are developed to a level comparable to that of open surgery, are now being performed to further assess laparoscopy's possible role in treating cancer. PMID- 9214325 TI - Elective laparoscopic-assisted sigmoid resection for diverticular disease. AB - BACKGROUND: Although the laparoscopic-assisted approach to colorectal cancer remains controversial, its use for benign diseases can have important advantages. The purpose of this study is to determine the feasibility of this approach for the treatment of elective diverticular disease and to identify preoperative and perioperative factors which can help to select the best procedure for each patient: either assisted laparoscopic resection (ALR) or dissection-facilitated laparoscopic resection (DLR). METHODS: From November 1991 to the present, we conducted a prospective study of 41 patients approached electively for diverticular disease. RESULTS: Twenty-nine patients underwent an ALR, seven were approached by DLR, and another five patients were converted to laparotomy (15%). Morbidity was 17.5% and there was no mortality in this series. The mean hospital stay after operation was 6.5 days. CONCLUSIONS: Because of the complexity of this inflammatory process, choice of either an assisted or a more invasive laparoscopic facilitated approach is necessary. The decision is based on the technical difficulty as determined by data collected both preoperatively and during laparoscopy. PMID- 9214326 TI - Laparoscopic-assisted colon surgery by abdominal wall lifting with newly developed lifting bars. AB - BACKGROUND: The aim of the study is to evaluate the efficacy of laparoscopic assisted colon surgery by lifting the abdominal wall with newly developed lifting bars. METHODS: We have made and used two kinds of lifting bars: type I and type T. Two I-type lifting bars are used in transverse colectomy and right hemicolectomy. One I-type lifting bar and one T-type bar are used in sigmoid colectomy and low anterior resection. After the intestine is dissected and the mesenterium is treated under laparoscopy, a small laparotomy wound about 4 to 6 cm long is made, and the intestine is pulled out of the body for extracorporeal anastomosis. RESULTS: The mean operating time was 153.8 +/- 51.9 min, and no particular complications were noted. CONCLUSIONS: Since postoperative pain is mild and postoperative recovery is rapid, this method is considered to be an effective surgical procedure. PMID- 9214327 TI - Application of rectal stents for palliation of obstructing rectosigmoid cancer. AB - BACKGROUND: The rationale of palliative endoscopic treatment is to avoid a colostomy in patients with advanced disease and limited life expectancy. This study was conducted to evaluate the role of endoscopic stent implantation for palliation of obstructing rectal cancer. METHODS: Overall, 19 patients (aged 47 87 years) with nonresectable or metastatic rectal cancer were treated by stent insertion after laser recanalization or dilation. Three types of stents, i.e., plastic tubes (n = 8), self-expanding mesh stents (n = 6), and endocoil stents (n = 5), were used to maintain luminal patency. RESULTS: Endoscopic stent implantation was successfully performed in all 19 patients. Long-term luminal patency and satisfactory bowel function were achieved in 16 of 19 patients (84%). After a median follow-up of 6 months, eight of the patients have died and eight are still alive without evidence of recurrent obstruction. Dislocation of the endoprosthesis occurred in two of eight plastic tubes and one of five mesh stents. Recurrent obstruction due to tumor ingrowth was only observed in patients treated with self-expanding mesh stents (n = 2). In spite of reinsertion and laser therapy a colostomy was required in three of 19 patients. There was no evidence of treatment failure in five patients who received endocoil stents. None of the patients experienced serious complications related to the endoscopic procedure. CONCLUSIONS: Endoscopic stent implantation seems to be a safe and efficient palliative approach to selected patients with obstructing rectal cancer. Currently, self-expanding coil stents are superior to other devices because of lower risk of dislocation and tumor ingrowth. PMID- 9214329 TI - Laparoscopic treatment of a benign splenic cyst. AB - With an understanding of the spleen's important immunologic function, splenectomy for benign splenic disorders has given way to a variety of splenic conservation techniques. Treatment options for benign nonparasitic splenic cysts include partial splenectomy, total cystectomy, or partial cyst decapsulation. External cyst wall decapsulation is a simplified operative procedure that carries no increased risk of cyst recurrence. However, a conventional upper abdominal laparotomy may subject patients to significant morbidity. We successfully performed a laparoscopic partial cyst decapsulation, achieving meticulous hemostasis with use of a laparoscopic-GIA stapling device. The patient tolerated the procedure well and was discharged on postoperative day 2. Follow-up has demonstrated no evidence of recurrent cyst formation. PMID- 9214328 TI - Laparoscopic colposuspension. Is it cost-effective? AB - BACKGROUND: The laparoscopic approach must be shown to be cost-effective as well as safe and technically effective before being widely adopted. A review of 54 consecutive patients who underwent open and laparoscopic colposuspension is presented and a cost-analysis is performed comparing the two approaches. METHODS: This study was a retrospective controlled review of patient records and accounts of in-hospital costs incurred at a private hospital. RESULTS: Theater costs were significantly greater in the laparoscopic group but this was balanced by a shorter length of stay and subsequent reduced accommodation cost. There was no difference in the overall in-hospital costs between the two groups. CONCLUSION: The laparoscopic surgical approach is safe and effective and by no means more expensive than the open approach. In the future, the laparoscopic approach can only become more cost efficient; techniques will improve and there will be earlier returns to work and, subsequently, greater productivity. PMID- 9214330 TI - Prosthetic reinforcement of posterior cruroplasty during laparoscopic hiatal herniorrhaphy. AB - Symptomatic gastroesophageal reflux after Nissen fundoplication may occur if the wrap herniates into the thorax. In an attempt to prevent recurrent hiatal hernia we employed polytetrafluoroethylene (PTFE) mesh reinforcement of posterior cruroplasty during laparoscopic Nissen fundoplication and hiatal herniorrhaphy. Three patients with symptomatic gastroesophageal reflux and a large (>==8 cm) hiatal defect underwent laparoscopic posterior cruroplasty and Nissen fundoplication. The cruroplasty was reinforced with a PTFE onlay. No perioperative complications occurred, and in follow-up (<==11 months) the patients are doing well. When repairing a large defect of the esophageal hiatus during fundoplication, the surgeon may consider reinforcement of the repair with PTFE mesh. PMID- 9214331 TI - Ileocutaneous fistula formation following laparoscopic polypropylene mesh hernia repair. AB - A rare case of enterocutaneous fistula caused by chronic erosion of polypropylene mesh after laparoscopic repair of a recurrent inguinal hernia is described. Successful treatment was achieved by fistulectomy, total resection of the implanted mesh, and small-bowel segmental resection. The patient recovered well postoperatively, and at follow-up 18 months later, the herniorrhaphy has remained intact. This complication needs to be added to the differential diagnosis in patients who present inflammation, abscess formation, or cutaneous fistula following laparoscopic hernia repair. PMID- 9214332 TI - Consequences of lost gallstone. AB - Laparoscopic cholecystectomy has become the treatment of choice in the management of calculus gallbladder disease. Intraperitoneal gallstone loss is not uncommon; it occurs in up to 40% of cases. Often, the stones are left unretrieved and are thought to be inconsequential. We present a series of patients who have had serious sequela from gallstones in the peritoneal cavity. We performed a retrospective study of the management of six patients with complications from intraperitoneal gallstones. The patients presented with a variety of complaints, from fevers to pneumonia to a colo-cutaneous fistula. Presentation ranged from immediately postoperatively to 18 months after surgery. Diagnosis included perihepatic abscesses and colo-biliary fistula. General anesthesia was usually necessary for removal of the stones. All patients have resolved following the removal of the gallstones. Our recommendation is to attempt to avoid spillage through careful dissection and retrieve any lost stones. The defect in the gallbladder can be closed with a clip. Whether the procedure should be converted to an open one to retrieve all the stones remains open to debate. The surgeon should be aware of the possible consequences of the lost gallstone. PMID- 9214334 TI - Endoscopically guided percutaneous repair of inguinal hernia through a 2-cm incision. Minihernia repair. AB - BACKGROUND: The laparoscopic repair of inguinal hernia is still controversial. Transabdominal preperitoneal repair violates the peritoneal cavity and may result in visceral injuries or intestinal obstruction. The laparoscopic extraperitoneal approach has the disadvantage of being technically demanding and requires extensive extraperitoneal mobilization. The Lichtenstein repair gives good long term results, is easy to learn, can be performed under local anesthesia, but requires a larger incision. METHODS: We describe a novel percutaneous tension free prosthetic mesh repair performed through a 2-cm groin incision. The inguinal canal is traversed with the aid of a 5-mm video-endoscope and the canal is widened using specially designed balloons. Spermatic cord mobilization, identification and excision of the indirect sac, and posterior wall repair are carried out under endoscopic guidance. RESULTS: Between October 1993 and July 1995, 85 primary inguinal hernia repairs (48 indirect and 33 direct) were performed on 81 patients (80 men, one woman) by the author (A.D.). The mean age was 41 years (range 17-83 years). Six repairs were performed under local anesthetic. Mean operative time was 42 min (range 25-74). Mean hospital stay was 1.2 days (0-3 days). The mean return to normal activity was 8 days (2-10 days). Eight complications have occurred: a serous wound discharge, two scrotal hematomas, a scrotal swelling that resolved spontaneously, wound pain lasting 2 weeks, an episode of urinary retention, and two recurrences early in the series (follow-up 1-22 months). CONCLUSION: The endoscopically guided percutaneous hernia repair avoids the disadvantages of laparoscopy (i.e., lack of stereoscopic vision, reduced tactile feedback, unfamiliar anatomical approach, risk of visceral injury), yet the use of endoscopic instrumentation allows operation through a 2-cm incision. The minihernia repair thus combines the virtues of an open tension-free repair with minimal access trauma. PMID- 9214335 TI - The "knot before loop" technique for the endoscopic ligation and suture instrument set. Knot formation and application. AB - BACKGROUND: In minimally invasive surgery intracorporal knot-tying is complicated by a limited field of vision and depth perception. METHODS: The "knot before loop" technique aims to reduce intraabdominal movements in number and space needed. A grasping instrument 3 mm in diameter guides a slipfit hollow knot pusher with a notch to hold the thread, when extracorporally forming the knot on the instrument tip and an axial slot. The loop is finished under endoscopic vision, yet a second loop is created along the thread. The knot is tightened and secured by closing the second loop without troublesome instrument change. RESULTS: The strength of the knot was tested and the feasibility of the instrument set was proven in pigs and 25 cholecystectomies and hernia repairs in humans. CONCLUSIONS: Endoscopic application of a secured slip knot is simplified by the "knot before loop" technique. The independent formation of the knot by the assisting personnel allows quick application, equivalent to the use of clips and staples. The benefit in cost saving is high. PMID- 9214375 TI - Study of biological properties of trichinella spiralis newborn larvae and the antiparasitic mucosal immunity of the host. AB - Studies of biological properties of newborn larvae of Trichinella spiralis and immune responses against newborn larvae in the host are reviewed. The biological properties of newborn larvae examined are the natural production, migration, surface antigens and their maturity and effects on the host. Various mechanisms of host immunity to newborn larvae are reviewed, including the stage specificity, kinetics, cell-mediated vs. antibody-dependent immunity in the mucosal and other tissues of the host. PMID- 9214377 TI - Human cytomegalovirus immediate early interaction with host nuclear structures: definition of an immediate transcript environment. AB - The development of an induced transcript environment was investigated at the supramolecular level through comparative localization of the human cytomegalovirus immediate early (IE) transcripts and specific nuclear domains shortly after infection. Compact aggregates of IE transcripts form only adjacent to nuclear domain 10 (ND10), and the viral protein IE86 accumulates exclusively juxtaposed to the subpopulation of ND10 with transcripts. The stream of transcripts is funneled from ND10 into the spliceosome assembly factor SC35 domain through the accumulation of IE86 protein, which recruits some components of the basal transcription machinery. Concomitantly the IE72 protein binds to ND10 and later disperses them. The domain containing the zinc finger region of IE72 is essential for this dispersal. Positional analysis of proteins IE86 and IE72, IE transcripts, ND10, the spliceosome assembly factor SC35, and basal transcription factors defines spatially and temporally an immediate transcript environment, the basic components of which exist in the cell before viral infection, providing the structural environment for the virus to usurp. PMID- 9214376 TI - Membrane dynamics at the endoplasmic reticulum-Golgi interface. PMID- 9214378 TI - A small conserved domain in the yeast Spa2p is necessary and sufficient for its polarized localization. AB - SPA2 encodes a yeast protein that is one of the first proteins to localize to sites of polarized growth, such as the shmoo tip and the incipient bud. The dynamics and requirements for Spa2p localization in living cells are examined using Spa2p green fluorescent protein fusions. Spa2p localizes to one edge of unbudded cells and subsequently is observable in the bud tip. Finally, during cytokinesis Spa2p is present as a ring at the mother-daughter bud neck. The bud emergence mutants bem1 and bem2 and mutants defective in the septins do not affect Spa2p localization to the bud tip. Strikingly, a small domain of Spa2p comprised of 150 amino acids is necessary and sufficient for localization to sites of polarized growth. This localization domain and the amino terminus of Spa2p are essential for its function in mating. Searching the yeast genome database revealed a previously uncharacterized protein which we name, Sph1p (a2p omolog), with significant homology to the localization domain and amino terminus of Spa2p. This protein also localizes to sites of polarized growth in budding and mating cells. SPH1, which is similar to SPA2, is required for bipolar budding and plays a role in shmoo formation. Overexpression of either Spa2p or Sph1p can block the localization of either protein fused to green fluorescent protein, suggesting that both Spa2p and Sph1p bind to and are localized by the same component. The identification of a 150-amino acid domain necessary and sufficient for localization of Spa2p to sites of polarized growth and the existence of this domain in another yeast protein Sph1p suggest that the early localization of these proteins may be mediated by a receptor that recognizes this small domain. PMID- 9214379 TI - Aminopeptidase I is targeted to the vacuole by a nonclassical vesicular mechanism. AB - The yeast vacuolar protein aminopeptidase I (API) is synthesized as a cytosolic precursor that is transported to the vacuole by a nonclassical targeting mechanism. Recent genetic studies indicate that the biosynthetic pathway that transports API uses many of the same molecular components as the degradative autophagy pathway. This overlap coupled with both in vitro and in vivo analysis of API import suggested that, like autophagy, API transport is vesicular. Subcellular fractionation experiments demonstrate that API precursor (prAPI) initially enters a nonvacuolar cytosolic compartment. In addition, subvacuolar vesicles containing prAPI were purified from a mutant strain defective in breakdown of autophagosomes, further indicating that prAPI enters the vacuole inside a vesicle. The purified subvacuolar vesicles do not appear to contain vacuolar marker proteins. Immunogold EM confirms that prAPI is localized in cytosolic and in subvacuolar vesicles in a mutant strain defective in autophagic body degradation. These data suggest that cytosolic vesicles containing prAPI fuse with the vacuole to release a membrane-bounded intermediate compartment that is subsequently broken down, allowing API maturation. PMID- 9214380 TI - Passive sorting in maturing granules of AtT-20 cells: the entry and exit of salivary amylase and proline-rich protein. AB - Previous studies have suggested that salivary amylase and proline-rich protein are sorted differently when expressed in AtT-20 cells (Castle, A.M., L.E. Stahl, and J.D. Castle. 1992. J. Biol. Chem. 267:13093- 13100; Colomer, V., K. Lal, T.C. Hoops, and M.J. Rindler. 1994.EMBO (Eur. Mol. Biol. Organ.) J. 13:3711- 3719). We now show that both exocrine proteins behave similarly and enter the regulated secretory pathway as judged by immunolocalization and secretagogue- dependent stimulation of secretion. Analysis of stimulated secretion of newly synthesized proline-rich protein, amylase, and endogenous hormones indicates that the exogenous proteins enter the granule pool with about the same efficiency as the endogenous hormones. However, in contrast to the endogenous hormones, proline rich protein and amylase are progressively removed from the granule pool during the process of granule maturation such that only small portions remain in mature granules where they colocalize with the stored hormones. The exogenous proteins that are not stored are recovered from the incubation medium and are presumed to have undergone constitutive-like secretion. These results point to a level of sorting for regulated secretion after entry of proteins into forming granules and indicate that retention is essential for efficient storage. Consequently, the critical role of putative sorting receptors for regulated secretion may be in retention rather than in granule entry. PMID- 9214381 TI - Multiple exocytotic pathways in pancreatic beta cells. AB - Ca2+-dependent exocytotic pathways in mouse pancreatic beta cells were investigated using both capacitance measurement and amperometric detection of vesicular contents. Serotonin was preloaded into large dense-core vesicles for the amperometry. Exocytosis was induced by rapid elevation of cytosolic Ca2+ concentrations using caged-Ca2+ compounds. Capacitance measurement revealed two major components of exocytosis, and only the slow component was accompanied by amperometric events reflecting quantal serotonin secretion. Moreover, the fast and slow exocytoses induced the two forms of endocytosis that were reported to follow the exocytoses of small-clear and large dense-core vesicles, respectively. Interestingly, we recorded two types of responses of quantal events: in the type 1 response, most quantal events occurred with a delay of 0.2 s and were rapidly exhausted with a time constant of 1.7 s, while, in the type-2 response, quantal events occurred with a delay of 2.5 s and were sustained. This suggests the existence of two pathways or modes of the exocytosis involving large dense-core vesicles. Thus, we have revealed three exocytotic pathways with divergent fusion kinetics in beta cells, which provide a new basis for the understanding of the physiology and pathology of beta cells. PMID- 9214382 TI - A novel class of RanGTP binding proteins. AB - The importin-alpha/beta complex and the GTPase Ran mediate nuclear import of proteins with a classical nuclear localization signal. Although Ran has been implicated also in a variety of other processes, such as cell cycle progression, a direct function of Ran has so far only been demonstrated for importin-mediated nuclear import. We have now identified an entire class of approximately 20 potential Ran targets that share a sequence motif related to the Ran-binding site of importin-beta. We have confirmed specific RanGTP binding for some of them, namely for two novel factors, RanBP7 and RanBP8, for CAS, Pse1p, and Msn5p, and for the cell cycle regulator Cse1p from Saccharomyces cerevisiae. We have studied RanBP7 in more detail. Similar to importin-beta, it prevents the activation of Ran's GTPase by RanGAP1 and inhibits nucleotide exchange on RanGTP. RanBP7 binds directly to nuclear pore complexes where it competes for binding sites with importin-beta, transportin, and apparently also with the mediators of mRNA and U snRNA export. Furthermore, we provide evidence for a Ran-dependent transport cycle of RanBP7 and demonstrate that RanBP7 can cross the nuclear envelope rapidly and in both directions. On the basis of these results, we propose that RanBP7 might represent a nuclear transport factor that carries an as yet unknown cargo, which could apply as well for this entire class of related RanGTP-binding proteins. PMID- 9214383 TI - Differential association of syntrophin pairs with the dystrophin complex. AB - The syntrophins are a multigene family of intracellular dystrophin-associated proteins comprising three isoforms, alpha1, beta1, and beta2. Based on their domain organization and association with neuronal nitric oxide synthase, syntrophins are thought to function as modular adapters that recruit signaling proteins to the membrane via association with the dystrophin complex. Using sequences derived from a new mouse beta1-syntrophin cDNA, and previously isolated cDNAs for alpha1- and beta2-syntrophins, we prepared isoform-specific antibodies to study the expression, skeletal muscle localization, and dystrophin family association of all three syntrophins. Most tissues express multiple syntrophin isoforms. In mouse gastrocnemius skeletal muscle, alpha1- and beta1-syntrophin are concentrated at the neuromuscular junction but are also present on the extrasynaptic sarcolemma. beta1-syntrophin is restricted to fast-twitch muscle fibers, the first fibers to degenerate in Duchenne muscular dystrophy. beta2 syntrophin is largely restricted to the neuromuscular junction. The sarcolemmal distribution of alpha1- and beta1-syntrophins suggests association with dystrophin and dystrobrevin, whereas all three syntrophins could potentially associate with utrophin at the neuromuscular junction. Utrophin complexes immunoisolated from skeletal muscle are highly enriched in beta1- and beta2 syntrophins, while dystrophin complexes contain mostly alpha1- and beta1 syntrophins. Dystrobrevin complexes contain dystrophin and alpha1- and beta1 syntrophins. From these results, we propose a model in which a dystrophin dystrobrevin complex is associated with two syntrophins. Since individual syntrophins do not have intrinsic binding specificity for dystrophin, dystrobrevin, or utrophin, the observed preferential pairing of syntrophins must depend on extrinsic regulatory mechanisms. PMID- 9214384 TI - In vitro reconstitution of cortical actin assembly sites in budding yeast. AB - We have developed a biochemical approach for identifying the components of cortical actin assembly sites in polarized yeast cells, based on a permeabilized cell assay that we established for actin assembly in vitro. Previous analysis indicated that an activity associated with the cell cortex promotes actin polymerization in the bud. After inactivation by a chemical treatment, this activity can be reconstituted back to the permeabilized cells from a cytoplasmic extract. Fractionation of the extract revealed that the reconstitution depends on two sequentially acting protein factors. Bee1, a cortical actin cytoskeletal protein with sequence homology to Wiskott-Aldrich syndrome protein, is required for the first step of the reconstitution. This finding, together with the severe defects in actin organization associated with the bee1 null mutation, indicates that Bee1 protein plays a direct role in controlling actin polymerization at the cell cortex. The factor that acts in the second step of the reconstitution has been identified by conventional chromatography. It is composed of a novel protein, Pca1. Sequence analysis suggests that Pca1 has the potential to interact with SH3 domain-containing proteins and phospholipids. PMID- 9214385 TI - A metastable intermediate state of microtubule dynamic instability that differs significantly between plus and minus ends. AB - The current two-state GTP cap model of microtubule dynamic instability proposes that a terminal crown of GTP-tubulin stabilizes the microtubule lattice and promotes elongation while loss of this GTP-tubulin cap converts the microtubule end to shortening. However, when this model was directly tested by using a UV microbeam to sever axoneme-nucleated microtubules and thereby remove the microtubule's GTP cap, severed plus ends rapidly shortened, but severed minus ends immediately resumed elongation (Walker, R.A., S. Inoue, and E.D. Salmon. 1989. J. Cell Biol. 108: 931-937). To determine if these previous results were dependent on the use of axonemes as seeds or were due to UV damage, or if they instead indicate an intermediate state in cap dynamics, we performed UV cutting of self-assembled microtubules and mechanical cutting of axoneme-nucleated microtubules. These independent methods yielded results consistent with the original work: a significant percentage of severed minus ends are stable after cutting. In additional experiments, we found that the stability of both severed plus and minus ends could be increased by increasing the free tubulin concentration, the solution GTP concentration, or by assembling microtubules with guanylyl-(alpha,beta)-methylene-diphosphonate (GMPCPP). Our results show that stability of severed ends, particularly minus ends, is not an artifact, but instead reveals the existence of a metastable kinetic intermediate state between the elongation and shortening states of dynamic instability. The kinetic properties of this intermediate state differ between plus and minus ends. We propose a three-state conformational cap model of dynamic instability, which has three structural states and four transition rate constants, and which uses the asymmetry of the tubulin heterodimer to explain many of the differences in dynamic instability at plus and minus ends. PMID- 9214387 TI - Translational diffusion of macromolecule-sized solutes in cytoplasm and nucleus. AB - Fluorescence recovery after photobleaching (FRAP) was used to quantify the translational diffusion of microinjected FITC-dextrans and Ficolls in the cytoplasm and nucleus of MDCK epithelial cells and Swiss 3T3 fibroblasts. Absolute diffusion coefficients (D) were measured using a microsecond-resolution FRAP apparatus and solution standards. In aqueous media (viscosity 1 cP), D for the FITC-dextrans decreased from 75 to 8.4 x 10(-7) cm2/s with increasing dextran size (4-2,000 kD). D in cytoplasm relative to that in water (D/Do) was 0.26 +/- 0.01 (MDCK) and 0.27 +/- 0.01 (fibroblasts), and independent of FITC-dextran and Ficoll size (gyration radii [RG] 40-300 A). The fraction of mobile FITC-dextran molecules (fmob), determined by the extent of fluorescence recovery after spot photobleaching, was >>0.75 for RG << 200 A, but decreased to <<0.5 for RG >> 300 A. The independence of D/Do on FITC-dextran and Ficoll size does not support the concept of solute "sieving" (size-dependent diffusion) in cytoplasm. Photobleaching measurements using different spot diameters (1.5-4 micron) gave similar D/Do, indicating that microcompartments, if present, are of submicron size. Measurements of D/Do and fmob in concentrated dextran solutions, as well as in swollen and shrunken cells, suggested that the low fmob for very large macromolecules might be related to restrictions imposed by immobile obstacles (such as microcompartments) or to anomalous diffusion (such as percolation). In nucleus, D/Do was 0.25 +/- 0.02 (MDCK) and 0.27 +/- 0.03 (fibroblasts), and independent of solute size (RG 40-300 A). Our results indicate relatively free and rapid diffusion of macromolecule-sized solutes up to approximately 500 kD in cytoplasm and nucleus. PMID- 9214386 TI - Control of mitotic events by Nap1 and the Gin4 kinase. AB - Little is known about the pathways used by cyclins and cyclin-dependent kinases to induce the events of the cell cycle. In budding yeast, a protein called Nap1 binds to the mitotic cyclin Clb2, and Nap1 is required for the ability of Clb2 to induce specific mitotic events, but the role played by Nap1 is unclear. We have used genetic and biochemical approaches to identify additional proteins that function with Nap1 in the control of mitotic events. These approaches have both identified a protein kinase called Gin4 that is required for the ability of Clb2 and Nap1 to promote the switch from polar to isotropic bud growth that normally occurs during mitosis. Gin4 is also required for the ability of Clb2 and Nap1 to promote normal progression through mitosis. The Gin4 protein becomes phosphorylated as cells enter mitosis, resulting in the activation of Gin4 kinase activity, and the phosphorylation of Gin4 is dependent upon Nap1 and Clb2 in vivo. Affinity chromatography experiments demonstrate that Gin4 binds tightly to Nap1, indicating that the functions of these two proteins are closely tied within the cell. These results demonstrate that the activation of Gin4 is under the control of Clb2 and Nap1, and they provide an important step towards elucidating the molecular pathways that link cyclin-dependent kinases to the events they control. PMID- 9214388 TI - Differential glycosylation of tractin and LeechCAM, two novel Ig superfamily members, regulates neurite extension and fascicle formation. AB - By immunoaffinity purification with the mAb Lan3-2, we have identified two novel Ig superfamily members, Tractin and LeechCAM. LeechCAM is an NCAM/FasII/ApCAM homologue, whereas Tractin is a cleaved protein with several unique features that include a PG/YG repeat domain that may be part of or interact with the extracellular matrix. Tractin and LeechCAM are widely expressed neural proteins that are differentially glycosylated in sets and subsets of peripheral sensory neurons that form specific fascicles in the central nervous system. In vivo antibody perturbation of the Lan3-2 glycoepitope demonstrates that it can selectively regulate extension of neurites and filopodia. Thus, these experiments provide evidence that differential glycosylation can confer functional diversity and specificity to widely expressed neural proteins. PMID- 9214389 TI - Stabilization of acetylcholine receptors by exogenous ATP and its reversal by cAMP and calcium. AB - Innervation of the neuromuscular junction (nmj) affects the stability of acetylcholine receptors (AChRs). A neural factor that could affect AChR stabilization was studied using cultured muscle cells since they express two distinct populations of AChRs similar to those seen at the nmjs of denervated muscle. These two AChR populations are (in a ratio of 9 to 1) a rapidly degrading population (Rr) with a degradation half-life of approximately 1 d and a slowly degrading population (Rs) that can alternate between an accelerated form (half life approximately 3-5 d) and a stabilized form (half-life approximately 10 d), depending upon the state of innervation of the muscle. Previous studies have shown that elevation of intracellular cAMP can stabilize the Rs, but not the Rr. We report here that in cultured rat muscle cells, exogenous ATP stabilized the degradation half-life of Rr and possibly also the Rs. Furthermore, pretreatment with ATP caused more stable AChRs to be inserted into the muscle membrane. Thus, in the presence of ATP, the degradation rates of the Rr and Rs overlap. This suggests that ATP released from the nerve may play an important role in the regulation of AChR degradation. Treatment with either the cAMP analogue dibutyryl cAMP (dB-cAMP) or the calcium mobilizer ryanodine caused the ATP-stabilized Rr to accelerate back to a half-life of 1 d. Thus, at least three signaling systems (intracellular cAMP, Ca2+, and extracellular ATP) have the potential to interact with each other in the building of an adult neuromuscular junction. PMID- 9214390 TI - Bi-transgenic mice reveal that K-rasVal12 augments a p53-independent apoptosis when small intestinal villus enterocytes reenter the cell cycle. AB - Studies in cell culture systems have indicated that oncogenic forms of Ras can affect apoptosis. Activating mutations of Ras occur in approximately 30% of all human tumors and 50% of colorectal carcinomas. Since these mutations appear at early or intermediate stages in multistep journeys to neoplasia, an effect on apoptosis may help determine whether initiated cells progress towards a more neoplastic state. We have tested the effects of K-rasVal12 on apoptosis in transgenic mice. A lineage-specific promoter was used to direct expression of human K-rasVal12, with or without wild-type (wt) or mutant SV-40 T antigens (TAg), in postmitotic villus enterocytes, the principal cell type of the small intestinal epithelium. Enterocytes can be induced to reenter the cell cycle by TAgWt. Reentry is dependent upon the ability of TAg to bind pRB and is associated with a p53-independent apoptosis. Analyses of K-rasVal12 x TAgWt bi-transgenic animals indicated that K-rasVal12 can enhance this apoptosis threefold but only in cycling cells; increased apoptosis does not occur when K-rasVal12 is expressed alone or with a TAg containing Glu107,108two head right arrow Lys107, 108 substitutions that block its ability to bind pRB. Analysis of bi-transgenic K rasVal12 x TAgWt mice homozygous for wild-type or null p53 alleles established that the enhancement of apoptosis occurs through a p53-independent mechanism, is not attributable to augmented proliferation or to an increase in abortive cell cycle reentry (compared to TAgWt mice), and is not associated with detectable changes in the crypt-villus patterns of expression of apoptotic regulators (Bcl 2, Bcl-xL, Bak, and Bax) or mediators of epithelial cell-matrix interactions and survival (e.g., alpha5beta1 integrin and its ligand, fibronectin). Coexpression of K-rasVal12 and TAgWt produces dysplasia. The K-rasVal12-augmented apoptosis is unrelated to this dysplasia; enhanced apoptosis is also observed in cycling nondysplastic enterocytes that produce K-rasVal12 and a TAg with a COOH-terminal truncation. The dysplastic epithelium of K-rasVal12 x TAgWt mice does not develop neoplasms. Our results are consistent with this finding: (a) When expressed in initiated enterocytes with a proliferative abnormality, K-rasVal12 facilitates progression to a dysplastic phenotype; (b) by diminishing cell survival on the villus, the oncoprotein may impede further progression; and (c) additional mutations may be needed to suppress this proapoptotic response to K-rasVal12. PMID- 9214391 TI - Involvement of ZO-1 in cadherin-based cell adhesion through its direct binding to alpha catenin and actin filaments. AB - ZO-1, a 220-kD peripheral membrane protein consisting of an amino-terminal half discs large (dlg)-like domain and a carboxyl-terminal half domain, is concentrated at the cadherin-based cell adhesion sites in non-epithelial cells. We introduced cDNAs encoding the full-length ZO-1, its amino-terminal half (N-ZO 1), and carboxyl-terminal half (C-ZO-1) into mouse L fibroblasts expressing exogenous E-cadherin (EL cells). The full-length ZO-1 as well as N-ZO-1 were concentrated at cadherin-based cell-cell adhesion sites. In good agreement with these observations, N-ZO-1 was specifically coimmunoprecipitated from EL transfectants expressing N-ZO-1 (NZ-EL cells) with the E-cadherin/alpha, beta catenin complex. In contrast, C-ZO-1 was localized along actin stress fibers. To examine the molecular basis of the behavior of these truncated ZO-1 molecules, N ZO-1 and C-ZO-1 were produced in insect Sf9 cells by recombinant baculovirus infection, and their direct binding ability to the cadherin/catenin complex and the actin-based cytoskeleton, respectively, were examined in vitro. Recombinant N ZO-1 bound directly to the glutathione-S-transferase fusion protein with alpha catenin, but not to that with beta catenin or the cytoplasmic domain of E cadherin. The dissociation constant between N-ZO-1 and alpha catenin was approximately 0.5 nM. On the other hand, recombinant C-ZO-1 was specifically cosedimented with actin filaments in vitro with a dissociation constant of approximately 10 nM. Finally, we compared the cadherin-based cell adhesion activity of NZ-EL cells with that of parent EL cells. Cell aggregation assay revealed no significant differences among these cells, but the cadherin-dependent intercellular motility, i.e., the cell movement in a confluent monolayer, was significantly suppressed in NZ-EL cells. We conclude that in nonepithelial cells, ZO-1 works as a cross-linker between cadherin/catenin complex and the actin-based cytoskeleton through direct interaction with alpha catenin and actin filaments at its amino- and carboxyl-terminal halves, respectively, and that ZO-1 is a functional component in the cadherin-based cell adhesion system. PMID- 9214392 TI - Direct Ca2+-dependent heterophilic interaction between desmosomal cadherins, desmoglein and desmocollin, contributes to cell-cell adhesion. AB - Human fibrosarcoma cells, HT-1080, feature extensive adherens junctions, lack mature desmosomes, and express a single known desmosomal protein, Desmoglein 2 (Dsg2). Transfection of these cells with bovine Desmocollin 1a (Dsc1a) caused dramatic changes in the subcellular distribution of endogenous Dsg2. Both cadherins clustered in the areas of the adherens junctions, whereas only a minor portion of Dsg2 was seen in these areas in the parental cells. Deletion mapping showed that intact extracellular cadherin-like repeats of Dsc1a (Arg1-Thr170) are required for the translocation of Dsg2. Deletion of the intracellular C-domain that mediates the interaction of Dsc1a with plakoglobin, or the CSI region that is involved in the binding to desmoplakin, had no effect. Coimmunoprecipitation experiments of cell lysates stably expressing Dsc1a with anti-Dsc or -Dsg antibodies demonstrate that the desmosomal cadherins, Dsg2 and Dsc1a, are involved in a direct Ca2+-dependent interaction. This conclusion was further supported by the results of solid phase binding experiments. These showed that the Dsc1a fragment containing cadherin-like repeats 1 and 2 binds directly to the extracellular portion of Dsg in a Ca2+-dependent manner. The contribution of the Dsg/ Dsc interaction to cell-cell adhesion was tested by coculturing HT-1080 cells expressing Dsc1a with HT-1080 cells lacking Dsc but expressing myc-tagged plakoglobin (MPg). In the latter cells, MPg and the endogenous Dsg form stable complexes. The observed specific coimmunoprecipitation of MPg by anti-Dsc antibodies in coculture indicates that an intercellular interaction between Dsc1 and Dsg is involved in cell-cell adhesion. PMID- 9214394 TI - Gene therapy for human renal disease? Don't catch your zinc finger in your leucine zipper. PMID- 9214395 TI - Clinical outcome relative to the dose of dialysis is not what you think: the fallacy of the mean. AB - Several recent retrospective studies of mortality relative to the dose of dialysis have been widely interpreted to indicate that adequate thrice-weekly hemodialysis requires a single pool Kt/V (spKt/V) of at least 1.4 to 1.6 and higher. In these studies, mortality rate has been correlated to the mean delivered spKt/V, (spKt/Vd)m, with coefficient of variation (CV) on the means ranging up to 45%. To evaluate these reported relationships, two large databases were analyzed using population constants to transform urea reduction ratio and spKt/Vd to equilibrated Kt/Vd (eKt/Vd), which expresses dose corrected for treatment time. The eKt/V dose (D) values were correlated to the reported relative risks (RR) of mortality to derive a RR/D function. The RR/D function, derived from these data with stepwise linear regression analysis, is nonlinear, with a steep linear increase in RR for eKt/Vd less than 1.05 and constant RR for eKt/Vd > or = 1.05. This RR/D function is mathematically expressed as RR = 1.96 - 1.03(eKt/Vd) (equation 1) when 0.50 < or = eKt/Vd < or = 1.05, and RR = 0.88 (equation 2) when eKt/V > or = 1.05. We show that regression of RR on (eKt/Vd)m with large CV results in overestimation of RR relative to eKt/Vd for individual patients because of extrapolation of the linear relationship beyond the threshold where the slope becomes zero (see equation 2 above). It is concluded that (1) current clinical data indicate that adequate dialysis is provided with eKt/Vd of 1.0 to 1.1 on a thrice-weekly schedule, (2) it is essential to assure that all patients achieve this level of therapy, which is best accomplished using urea kinetic modeling for both prescription and measurement of delivered eKt/Vd, and (3) the current HEMO study is well designed to determine whether higher levels of eKt/Vd will further improve clinical outcome. PMID- 9214396 TI - Race and creatinine excretion in chronic renal insufficiency. AB - Black race and the absence of diabetes are associated with higher levels of serum creatinine in patients with end-stage renal disease. We examined whether these factors have a similar influence on creatinine excretion in men with chronic renal insufficiency. The hypotheses were tested in one sample (group A, n = 35) and the findings replicated in a second, independent sample (group B, n = 66). Creatinine excretion normalized to weight (UCr/kg) was compared by race and diabetic status. UCr/kg and creatinine clearance also were compared with the values predicted by the Cockcroft-Gault (CG) formula (based on the regression equation, UCr/kg = 28 - age/5). In each sample, mean UCr/kg was significantly higher in black patients than in nonblack patients (group A, P = 0.004; group B, P = 0.029), and UCr/kg and creatinine clearance were significantly underestimated by the CG predictions in black patients (group A, P < or = 0.004; group B, P < or = 0.019), but not in nonblack patients. Diabetes did not significantly influence UCr/kg. The analysis also was performed at two age levels (< 50 years or > or = 50 years) using groups A and B combined. For black patients younger than 50 (n = 10), observed UCr/kg (P = 0.059) and creatinine clearance (P = 0.025) exceeded the values predicted by the CG formula; the analysis of nonblack patients younger than 50 years was limited by sample size (n = 1). For patients aged 50 years and older (black, n = 32; nonblack, n = 58), mean UCr/kg was significantly higher in black patients (P = 0.034), and UCr/kg and creatinine clearance were significantly underestimated by the CG predictions in black patients (P < or = 0.002) but not in nonblack patients. In multiple regression analysis of all patients aged 50 years and older, UCr/kg was independently influenced by both race (P < 0.05) and age (P < 0.04) (overall model, multiple R = 0.31; P = 0.012). The prediction equation was UCr/kg (mg/kg) = 23.6 - age/8.3 + 1.9 x race (race = 0 if nonblack; race = 1 if black). We conclude that the creatinine excretion rate was strongly affected by race but not diabetes in men with chronic renal insufficiency. The CG formula significantly underestimated UCr/kg and therefore creatinine clearance in black patients. These findings may reflect differences between black and nonblack subjects in body composition, muscle metabolism, or diet, and the interaction of these factors with chronic renal insufficiency. PMID- 9214393 TI - CDO: an oncogene-, serum-, and anchorage-regulated member of the Ig/fibronectin type III repeat family. AB - Cell adhesion molecules of the Ig superfamily are implicated in a wide variety of biological processes, including cell migration, axon guidance and fasciculation, and growth control and tumorigenesis. Expression of these proteins can be highly dynamic and cell type specific, but little is known of the signals that regulate such specificity. Reported here is the molecular cloning and characterization of rat CDO, a novel cell surface glycoprotein of the Ig superfamily that contains five Ig-like repeats, followed by three fibronectin type III-like repeats in its extracellular region, and a 256-amino acid intracellular region that does not resemble other known proteins. In rat embryo fibroblasts, cdo mRNA expression is maximal in confluent, quiescent cells. It is rapidly and transiently down regulated by serum stimulation of such cells, and is constitutively down regulated in oncogene-transformed derivatives of these cells. CDO protein levels are also dramatically regulated by cell-substratum adhesion, via a mechanism that is independent of cdo mRNA expression. The amount of CDO produced at the surface of a cell may therefore be governed by a complex balance of signals, including mitogenic stimuli that regulate cdo mRNA levels, and substratum-derived signals that regulate CDO protein production. cdo mRNA is expressed at low levels in most adult rat tissues. A closely related human gene maps to chromosome 11q23-24, a region that displays frequent loss of heterozygosity in human lung, breast, and ovarian tumors. Taken together, these data suggest that loss of CDO function could play a role in oncogenesis. PMID- 9214397 TI - Elevated plasma concentrations of neuropeptide Y in children and adults with chronic and terminal renal failure. AB - Neuropeptide Y (NPY) is a peptide hormone that is expressed, stored, and released in sympathetic neurones together with noradrenaline. Elevated plasma concentrations of NPY have been reported in patients with neural crest tumors (neuroblastoma, pheochromocytoma) and following exercise. We studied plasma concentrations of NPY in children and adults with chronic and terminal renal failure and compared them with those in healthy controls. Neuropeptide Y was significantly higher in children and adolescents receiving peritoneal dialysis (5.3 +/- 2.8 pmol/L; n = 11 [mean +/- SD]) or hemodialysis (5.4 +/- 2.1 pmol/L; n = 14) than in healthy children (2.3 +/- 0.9 pmol/L; n = 19) or pediatric patients with impaired renal function who are not receiving dialysis (2.7 +/- 0.6 pmol/L; n = 8; mean glomerular filtration rate, 41 mL/min x 1.73 m2). There was a small but insignificant negative correlation between glomerular filtration rate and NPY concentrations in children with impaired renal function (r = 0.49; P = 0.25). In healthy adults, NPY concentration was similar to that in healthy children (1.8 +/ 1.0 pmol/L; n = 13), and it was significantly elevated in adults receiving hemodialysis (5.9 +/- 1.7 pmol/L; n = 16). No significant changes in NPY concentrations were found before and after hemodialysis in pediatric or adult patients. We conclude that plasma concentrations of NPY are elevated in patients with chronic renal failure who are receiving either peritoneal or hemodialysis, but not in patients with moderately impaired renal function. Whether elevated NPY concentration indicates increased sympathetic activity or is caused by reduced NPY clearance remains to be shown. PMID- 9214398 TI - Severity of AIDS and the response to EPO in uremia. AB - To determine the factors that govern their response to erythropoietin (EPO), we conducted a cross-sectional study of all patients in four outpatient hemodialysis facilities in Brooklyn, NY, who had end-stage renal disease (ESRD) and human immunodeficiency virus (HIV) infection and were receiving recombinant EPO. We also compared the hematocrit and EPO requirements of these patients to those of a control group of hemodialysis patients without HIV infection. We documented known duration of HIV infection, and total CD4 count was measured once. In both groups, hematocrit was measured weekly for 5 weeks and a mean value calculated for each subject. Transferrin saturation was measured twice and a mean value calculated for each subject. Intensity of hemodialysis was assessed by measuring both percent reduction of urea and serum albumin concentration twice; mean values were calculated for each subject. Twenty-nine (88%) of 33 study subjects had acquired immunodeficiency syndrome. Mean known duration of HIV infection was 49 +/- 32.5 months (median, 48 months), and mean total CD4 count was 143 +/- 152.4 cells/mm3 (median, 72 cells/mm3). Mean hematocrit in the study subjects was 27.4% +/- 4.7% compared with 27.6% +/- 3.7% in the controls (P = 0.69). Mean thrice-weekly EPO dose was higher in the study subjects (90 +/- 52 U/kg body weight) than in the controls (62 +/- 36 U/Kg body weight) (P = 0.001). Among the study subjects, hematocrit had direct univariate correlations with serum albumin concentration (r = 0.43; P = 0.02), transferrin saturation (r = 0.4; P = 0.03), and percent reduction of urea (r = 0.4; P = 0.02), but not with total CD4 count (r = -0.05; P = 0.8) or known duration of HIV infection (r = -0.11; P = 0.55). There was an inverse correlation between hematocrit and dose of EPO (r = -0.5; P = 0.003). Multiple regression analysis showed that transferrin saturation (P = 0.01) and percent reduction of urea (P = 0.003) had direct correlations with hematocrit after adjustment for other factors. There was an inverse relationship between hematocrit and dose of EPO (P = 0.0006). We conclude that in patients with ESRD and HIV infection receiving hemodialysis, the response to EPO (hematocrit) is modulated by the dose of EPO, quantity of hemodialysis, and transferrin saturation, but not by the severity of HIV disease. Hemodialysis patients infected with HIV receive a higher dose of EPO than those without HIV infection. PMID- 9214399 TI - Temporal changes and reversibility of carbamylated hemoglobin in renal failure. AB - The detection of carbamylated hemoglobin (CarHb) is known to be useful in determination of the chronicity of uremia. However, the time course of the in vivo reaction between isocyanic acid and terminal valine residues of the hemoglobin chain is not clearly defined. To assess the temporal relationship and reversibility of carbamylation, we prospectively measured CarHb as micrograms of valine hydantoin per gram of hemoglobin (microg VH/g Hb) by high-performance liquid chromatography in 37 patients with acute renal failure (ARF), 53 patients with chronic renal failure (CRF), and six patients with successful kidney transplant. Patients with ARF had a lower median CarHb concentration (53.2 microg VH/g Hb; range, 24.6 to 97.1 microg VH/g Hb) than those with CRF (115.0 microg VH/g Hb; range, 34.6 to 286.5 microg VH/g Hb; P < 0.01), but had a higher value (53.2 microg VH/g Hb; range, 24.6 to 97.1 microg VH/g Hb) than 31 normal controls (36.6 microg VH/g Hb; range, 19.9 to 62.9 microg VH/g Hb; P < 0.05). In patients with ARF, the CarHb concentration positively correlated with the number of days of illness (r = 0.74; P < 0.01). The patients with ARF of 10 or more days' duration had a higher CarHb concentration (68.7 microg VH/g Hb; range, 36.0 to 93.9 microg VH/g Hb) than those with a shorter duration of ARF (33.7 microg VH/g Hb; range, 24.6 to 55.8 microg VH/g Hb; P < 0.01) despite similar blood urea nitrogen and serum creatinine values. However, they had a lower concentration of CarHb (68.7 microg VH/g Hb; range, 36.0 to 93.9 microg VH/g Hb) than CRF patients with comparable serum creatinine values (112.5 microg VH/g Hb; range, 34.6 to 286.5 microg VH/g Hb; P < 0.01). In patients with a kidney transplant, CarHb concentration declined by 19.7% (range, 12.3% to 35.6%) within 2 to 3 weeks after receiving the graft, while the level of hemoglobin increased by 25% (range, 4.0% to 46.6%) during the same period. Therefore, the total blood CarHb (CarHb x hemoglobin concentration) was not significantly changed. We concluded that the in vivo reaction of carbamylation of hemoglobin progressed during the period of uremia, and there might exist some irreversible preformed CarHb in advanced stages of CRF. PMID- 9214400 TI - Prevalence, recognition, and implications of mental impairment among hemodialysis patients. AB - The increasing age and co-morbidity of dialysis patients may be associated with an increase in the prevalence of Alzheimer's disease, stroke, and other causes of mental impairment. We sought to determine the prevalence, recognition, and implications of mental impairment among chronic hemodialysis patients. We administered the Mini Mental Status Exam (MMSE) to 336 randomly selected patients from three dialysis units. To determine recognition of mental impairment by health care providers, we compared MMSE scores with mental status assessments obtained from each patient's dialysis technician and medical record. To determine the clinical implications of mental impairment, we prospectively obtained Kt/V, albumin, protein catabolic rate, blood pressure, and hematocrit values. To determine the resource implications of mental impairment, we assessed staff time required to care for each patient as well as hospitalizations. We found that 22% of subjects had mild mental impairment (MMSE 18 to 23) and that 8% had moderate severe mental impairment (MMSE 0 to 17). The sensitivity of technician and medical record mental status assessments were 57% and 15%, respectively. After adjusting for demographic and medical variables, low MMSE score was independently associated with low protein catabolic rate (odds ratio, 1.5; P = 0.02), increased technician time caring for patient after dialysis (odds ratio, 1.5; P = 0.005), and increased hospital days (odds ratio, 1.4; P = 0.03). In conclusion, there is a high prevalence of unrecognized mental impairment among hemodialysis patients that has adverse implications for protein nutritional status, staff time, and hospitalization. We recommend that clinicians routinely screen for mental impairment and target impaired patients for interventions to improve mental status and associated adverse outcomes. PMID- 9214401 TI - Vascular access survival among incident hemodialysis patients in the United States. AB - Vascular access failure causes substantial morbidity to hemodialysis patients. We sought to identify factors determining survival of the permanent vascular access in use at the start of end-stage renal disease during 1990 in a national sample of 784 incident hemodialysis patients insured by Medicare. Medicare claims records were used to identify access failures or revisions among patients with an arteriovenous (AV) fistula (n = 245) and an AV vascular graft (n = 539). A proportional hazards analysis of time to first failure or revision, controlled by stratification for sex, race, and cause of end-stage renal disease, was used to determine the effect of age, access type, and peripheral vascular disease on vascular access survival. Patients with an AV fistula and who were older than 65 years had a risk of access failure that was 24% lower than similar patients with an AV graft (P < 0.02). The relative risk of access failure for an AV fistula, but not an AV graft, varied significantly with age for patients younger than 65 years (P < 0.01). The relative risk of access failure for a patient with an AV fistula, compared with a patient of the same age with an AV graft, was 67% lower at the age of 40 years, 54% lower at the age of 50 years, and 24% lower at the age of 65 years. A history of peripheral vascular disease was associated with a 24% higher risk of AV graft or fistula failure (P = 0.05). Measures to decrease vascular access-related morbidity among hemodialysis patients should include reversing the current trend toward increasing use of AV grafts, particularly in patients younger than 65 years. PMID- 9214402 TI - Blood volume changes during three different profiles of dialysate sodium variation with similar intradialytic sodium balances in chronic hemodialyzed patients. AB - The aim of this study was to evaluate the effects on blood volume (BV) preservation of three different profiles of dialysate sodium variation with similar intradialytic sodium balances. Ten uremic patients aged 50 +/- 11 years receiving regular bicarbonate hemodialysis for 49 +/- 57 months were studied. Each patient underwent three hemodialysis treatments with different modalities of dialysate sodium profiles: constant sodium hemodialysis (CHD), high-low sodium hemodialysis (H-LHD), and low-high sodium hemodialysis (L-HHD). In CHD, the dialysate sodium concentration was 141 mEq/L and did not change during treatment. In H-LHD and L-HHD, the dialysate sodium concentration at the start of dialysis was 160 mEq/L and 133 mEq/L, respectively, and remained constant for 60 minutes. At this time, a single-step break point of variation of dialysate sodium concentration occurred. The dialysate sodium concentration changed according to a model aimed to keep identical the amount of dialysate sodium exchanged in the three different dialysis procedures. The duration of hemodialysis, the blood flow rate, the dialysate flow rate, and the dialysis membrane were the same for all three different hemodialysis modalities. The ultrafiltration rate was kept constant during treatment. Total dialysate collection and intradialytic sodium balance were calculated for each hemodialysis session. Blood pressure and heart rate were monitored at 10-minute intervals; percent reductions of BV (%R-BV) were continuously monitored by an online optical reflection method (Hemoscan; Hospal Dasco, Medolla, Italy). The results have shown a lower intradialytic %R-BV with H LHD compared with L-HHD and CHD. No differences in total ultrafiltration rate, systolic and diastolic blood pressures, and heart rate were observed among the three different dialysis procedures. The total dialysate sodium collected and the intradialytic sodium balances were very similar among the three different dialysis procedures, confirming the accuracy of the precision of the sodium model used. The H-LHD sodium profile may be a useful tool in the prevention of excessive %R-BV and of dialysis intolerance episodes. PMID- 9214403 TI - Low levels of spontaneously activated peripheral IgA-secreting cells in nontransplanted IgA nephropathy patients. AB - The pathognomonic presence of IgA in the glomerular mesangium of patients with IgA nephropathy (IgAN) suggests an abnormal control of IgA metabolism in this disease that could be modified in transplanted IgAN patients. Alterations of IgA production have been demonstrated in both bone marrow samples and mucosae associated tissues from IgAN patients, but the analysis of IgA production by peripheral B lymphocytes in culture has yielded conflicting results, some investigators showing an enhanced secretion of IgA and others reporting similar data as with control samples. Little is known about the endogenous activation state of B cells in IgAN, which can be approached by analyzing peripheral Ig producing cells, a number of which are recirculating between lymphoid organs. In the present study, two methods were applied to appreciate the numbers of spontaneously activated peripheral B cells. The ELISPOT method provided information about short-term secretion of Igs. Intracytoplasmic immunofluorescence for IgA allowed to identify IgA-containing cells, either as short-rimmed immunocytes or as brightly stained immunoblasts with an enlarged cytoplasm. These cells were enumerated in IgAN patients (n = 31), transplanted IgAN patients (n = 27), control patients with other biopsy-proven renal diseases (nontransplanted, n = 48; transplanted, n = 38), and in healthy individuals (n = 18). The number of IgA spot-forming cells obtained in the control groups (1,251 +/- 95 cells/10(6) lymphocytes [mean +/- SE]) was consistent with those in similar previously reported studies, but differed significantly (P = 0.01) from those observed in nontransplanted IgAN patients, who had a surprisingly lower number of such cells (699 +/- 97 cells/10(6) lymphocytes); the number of IgA spot forming cells in the transplanted IgAN patients (1,355 +/- 182 cells/10(6) lymphocytes) did not differ from that in the control groups. The same pattern was seen for IgA-containing immunoblasts. There was no difference in IgG (overall, 214 +/- 13 cells/10(6) lymphocytes) and IgM (overall, 61 +/- 10 cells/10(6) lymphocytes) spot-forming cell numbers between the five groups of individuals tested, suggesting that the anomaly noted in IgAN patients was not related to technical problems and that this could be a new feature of this renal disease. The normal levels of spontaneously activated peripheral IgA-producing cells found in transplanted IgAN patients suggest that immunosuppressive treatments could interfere with the anomalies of IgA metabolism in this disease. PMID- 9214404 TI - Sodium depletion enhances fibrosis and the expression of TGF-beta1 and matrix proteins in experimental chronic cyclosporine nephropathy. AB - The major limitation to the clinical use of cyclosporine (CsA) is renal toxicity. In the past, the lack of an animal model of chronic CsA nephropathy has hampered the study of its pathogenesis. Rats given CsA and placed on a low sodium diet (LSD) develop a histology similar to human lesions of chronic CsA nephropathy, a phenomenon not observed in animals on a normal sodium diet (NSD). We have previously shown that transforming growth factor-beta1 (TGF-beta1) is involved in the CsA-induced renal fibrosis in rats on a LSD. We hypothesized that sodium depletion is critical to the increase in TGF-beta1 expression, which, in turn, results in excessive matrix accumulation. Pair-fed rats were placed on a NSD or LSD, treated with CsA or vehicle, and killed at 7 or 28 days (N = 4 to 6 in each group). All rats achieved similar blood pressure control, and all CsA-treated rats achieved similar CsA blood levels. However, while CsA did not affect creatinine clearance in rats on a NSD, it lowered creatinine clearance in rats on a LSD (P < 0.01). Cyclosporine-induced tubulointerstitial fibrosis and arteriolopathy was observed at 28 days only in the rats on a LSD (P < 0.05). In addition, peripheral renin activity was increased only in the rats on a LSD (P < 0.01), while it remained normal in the rats on a NSD. In addition, CsA-treated rats on a LSD developed a progressive increase in the mRNA expression of TGF beta1 and the matrix proteins biglycan and type I collagen at 7 and 28 days. Most of the changes were seen at 28 days (P < 0.001 for TGF-beta1, P < 0.01 for biglycan and type I collagen). On the other hand, CsA treatment in rats on a NSD did not affect the mRNA expression of TGF-beta1 and matrix proteins. Most of the changes in the immunofluorescence deposition of the glycoproteins tenascin and fibronectin EDA+ were in the tubulointerstitium and vessels of the kidneys of rats on a LSD and were mostly significant at 28 days, in accordance with the characteristic histology of chronic CsA nephropathy. The mRNA expression of plasminogen activator inhibitor-1, a protease inhibitor involved in matrix degradation and stimulated by TGF-beta1, was observed only in kidneys of rats on a LSD (P < 0.01). Since sodium depletion elevates peripheral renin activity, our experiments suggest a role for the renin-angiotensin system in the expression of TGF-beta1 and matrix proteins in CsA-induced renal fibrosis of rats on a LSD. PMID- 9214405 TI - Endothelin and atrial natriuretic peptide levels following radiocontrast exposure in humans. AB - Radiocontrast exposure is associated with vasoconstriction of the renal vascular bed and, in certain circumstances, with acute renal failure. This may be influenced by the volume of contrast infused or underlying disease, such as diabetes or renal failure. Changes in circulating vascular regulators, such as endothelin and atrial natriuretic peptide (ANP), may play a role in the development and/or prevention of acute renal failure. Nineteen patients undergoing arteriographic procedures were divided into two groups: large-volume contrast (> or = 150 mL; n = 7) and small-volume contrast (< 150 mL; n = 12). Circulating endothelin levels increased significantly (from 12.3 +/- 1.1 pmol/L to 19.4 +/- 2.2 pmol/L; P < 0.05) following large-volume contrast exposure (group 1) but not following small-volume contrast exposure (group 2) (13.9 +/- 1.7 pmol/L to 12.2 +/- 0.09 pmol/L). ANP levels increased significantly in both groups: 43 +/- 15 pg/mL to 75 +/- 21 pg/mL in group 1 and 33 +/- 16 to 106 +/- 39 pg/mL in group 2. Data from an additional eight patients with underlying diabetes mellitus and/or renal insufficiency also were obtained and were considered separately. Endothelin levels were higher at baseline and increased significantly after contrast (25.7 +/- 5 pmol/L to 55.4 +/- 18 pmol/L) despite the relatively small average volume of contrast infused (112 +/- 15 mL). ANP levels were also highest in these patients (211 +/- 43 pg/mL precontrast and 323 +/- 65 pg/mL postcontrast). No group had a significant change in serum creatinine following contrast exposure. In conclusion, large-volume radiocontrast exposure is associated with an increase in both circulating endothelin and ANP levels. Patients with underlying diabetes or renal insufficiency may have higher baseline levels and a greater tendency to increase endothelin after contrast exposure. While an increase in endothelin may contribute to renal vasoconstriction following radiocontrast exposure, simultaneous increases in ANP may serve to offset this response and protect against changes in renal function. PMID- 9214406 TI - Demonstration of the proliferation marker Ki-67 in renal biopsies: correlation to clinical findings. AB - Assessment of cell proliferation in renal biopsy samples is a potentially promising analytical tool to evaluate disease activity. So far no information is available on the correlation between proliferative activity in different anatomic compartments of the kidney and clinical symptoms. To elucidate this issue, we examined renal biopsy specimens from 20 patients with systemic vasculitis (15 Wegener's granulomatosis, five microscopic polyangiitis), 20 patients with immunoglobulin (Ig) A nephropathy (IgAN), 13 patients with minimal-change disease (MCD), 11 patients with tubulointerstitial nephritis, and five patients with diabetes mellitus. The streptavidin-biotin-peroxidase complex technique was applied to autoclave-pretreated, formalin-fixed, paraffin-embedded tissue sections to label different cell types with the antibody MIB1 directed against the Ki-67 antigen. Proliferation index (PI) was estimated as the number of positively stained nuclei per glomerular cross-section or per square millimeter section area. The interstitial cells were discriminated by additional staining of Ki-67-processed samples with specific immune markers. In patients with vasculitis, PI was considerably elevated in the extracapillary glomerular compartment (0.86), in proximal tubules (6.24), and in the interstitium (8.62). High proliferative activity was also noted in interstitium (3.98) and proximal tubules (1.35) of patients with IgAN. Of particular interest was the increased interstitial proliferative activity (15.0) in diabetic patients. Resident renal cells, but not infiltrating cells, seemed to constitute the majority of the proliferating cell population in the interstitium. In systemic vasculitis, clinical disease activity was significantly correlated to endocapillary (r(s) = 0.58), extracapillary (r(s) = 0.67), proximal tubular (r(s) = 0.67), and interstitial PI (r(s) = 0.61). By multiple linear regression analysis, proximal tubular PI was correlated to the presence of hematuria (beta = 0.72) and to interstitial fibrosis score (beta = 0.59). Interstitial PI was independently correlated to antineutrophil cytoplasmic antibodies (ANCA) titer (beta = 0.7) and interstitial fibrosis score (beta = 0.55), and it was the only one PI correlated to serum creatinine concentration (beta = 0.53). The independent association between interstitial PI and serum creatinine (beta = 0.64) was also found in IgAN. Proximal tubular PI was correlated to interstitial fibrosis score (beta = 0.59) and proteinuria (beta = 0.54). In MCD, high PI values were noted in proximal tubular cells (1.42) but not in glomeruli and the interstitium. In conclusion, assessment of proliferation activity by immunohistology provides additional information beyond conventional pathological techniques to evaluate disease activity and prognosis in renal biopsies. PMID- 9214407 TI - Elevated cytosolic calcium and impaired proliferation of B lymphocytes in type II diabetes mellitus. AB - Patients with diabetes mellitus have increased susceptibility to infection attributable, at least in part, to defective function of polymorphonuclear leukocytes (PMNLs) and B cells. Certain data suggest that cytosolic calcium ([Ca2+]i) is elevated in various cells in diabetes, and high [Ca2+]i adversely affects cell function. Indeed, the [Ca2+]i of PMNLs of diabetic patients is elevated, and phagocytosis of the PMNLs is impaired. The current study examines whether the [Ca2+]i of B cells is also elevated in diabetes and whether this derangement impairs B cell function. We studied 32 patients with non-insulin dependent diabetes mellitus (NIDDM) and eight normal subjects. All patients had hyperglycemia (11.6 +/- 0.80 mmol/L) and elevated HbA1c (13.2% +/- 0.99%). The basal levels of [Ca2+]i of the B cells (113 +/- 3.3 nmol/L) were significantly (P < 0.01) higher than the values in normal subjects (85 +/- 1.7 nmol/L). There was a direct and significant correlation (r = 0.88; P < 0.01) between the [Ca2+]i of B cells and the blood levels of glucose. Proliferation of B cells in response to Staphylococcus aureus Cowan I (SAC) was significantly impaired in these patients (7.3 +/- 0.48 x 10(3) cpm v 12.5 +/- 0.61 x 10(3) cpm in normal subjects). Normalization of blood glucose with the hypoglycemic agents, glyburide, was associated with the return of both [Ca2+]i of B cells and their proliferation in response to SAC to normal. The results show that hyperglycemia in type II diabetes mellitus is associated with a significant increase in [Ca2+]i of B cells and with a decrease in their proliferation in response to mitogen. These derangements are reversed after the correction of the hyperglycemia. The data of the current study and those previously reported in PMNLs provide for a new pathogenetic process underlying the dysfunction of these cells in diabetes mellitus. PMID- 9214409 TI - Response to high-dose interferon-alpha after failure of standard therapy in MPGN associated with hepatitis C virus infection. AB - A 42-year-old man developed mixed cryoglobulinemia secondary to hepatitis C virus (HCV) infection with hypocomplementemia and nephrotic syndrome. His renal biopsy showed membranoproliferative glomerulonephritis type I (MPGN). Despite treatment with interferon-alpha, three million units three times a week for a total of 6 months, the patient continued to have hypocomplementemia, cryoglobulinemia, and nephrosis. After a course of high-dose interferon-alpha treatment consisting of ten million units daily for 2 weeks followed by 10 million units three times per week for an additional 6 weeks, HCV RNA and cryoglobulin testing became negative, complement levels increased to normal levels, and nephrotic syndrome remitted. This case confirms an association between HCV infection and MPGN and suggests a role for high-dose interferon-alpha treatment when conventional interferon therapy fails. PMID- 9214408 TI - A new analog of 1,25-(OH)2D3, 19-NOR-1,25-(OH)2D2, suppresses serum PTH and parathyroid gland growth in uremic rats without elevation of intestinal vitamin D receptor content. AB - We have previously reported that 19-nor-1,25-(OH)2D2, a new analog of 1,25 (OH)2D3, suppresses parathyroid hormone (PTH) secretion in uremic rats in the absence of hypercalcemia or hyperphosphatemia. In the current study, we examined the effect of 19-nor-1,25-(OH)2D2 on parathyroid gland growth and intestinal vitamin D receptor (VDR) content. After induction of uremia by 5/6 nephrectomy, rats were divided into five experimental groups and received intraperitoneal injections of vehicle, 1,25-(OH)2D3 (2 or 6 ng/rat), or 19-nor-1,25-(OH)2D2 (25 or 100 ng/rat) three times a week for 8 weeks. Twelve normal rats received vehicle and served as the normal control group. During the course of the study, rats were maintained on a 1.0% calcium and 0.8% phosphorus diet. The higher dose of 1,25-(OH)2D3, 6 ng, significantly decreased PTH from 52.7 +/- 10.2 pg/mL in the uremic control group to 25.7 +/- 6.7 pg/mL (P < 0.01). This dose of 1,25 (OH)2D3, however, increased serum levels of both ionized calcium (4.71 +/- 0.05 to 4.85 +/- 0.06 mg/dL; P < 0.05) and phosphorus (4.34 +/- 0.30 to 6.67 +/- 0.63 mg/dL; P < 0.01). Both doses of 19-nor-1,25-(OH)2D2 decreased serum PTH as effectively as 1,25-(OH)2D3 without changes in serum calcium or phosphorus. The 100-ng dose of 19-nor-1,25-(OH)2D2 decreased PTH to 20.7 +/- 3.1 pg/mL (P < 0.01) and suppressed parathyroid gland growth by more than 50%. Both doses of 19-nor 1,25-(OH)2D2 also decreased endogenous 1,25-(OH)2D3 levels compared with uremic control rats (25 ng:30.4 +/- 2.0, P < 0.05, and 100 ng:27.9 +/- 3.2, P < 0.01, v 48.4 +/- 6.6 pg/mL). The 6-ng dose of 1,25-(OH)2D3 elevated intestinal VDR content (138.5 +/- 20.0 fmol/mg protein) compared with animals receiving both doses of 19-nor-1,25-(OH)2D2 (25 ng:84.0 +/- 11.9, P < 0.05, and 100 ng:78.4 +/- 10.9, P < 0.01). This was probably attributable to the marked decrease in endogenous 1,25-(OH)2D3 levels caused by both doses of 19-nor-1,25-(OH)2D2 because intestinal VDR correlated directly with serum 1,25-(OH)2D3 (r = 0.963; P = 0.008). Thus, 19-nor-1,25-(OH)2D2 appears to exert a selective action on the parathyroid glands compared with the intestine. Its low calcemic and phosphatemic properties may result from the decreased endogenous 1,25-(OH)2D3 levels that lead to a reduction in intestinal VDR. This selectivity makes this analog ideal for the treatment of secondary hyperparathyroidism. PMID- 9214410 TI - AIDS-associated membranous nephropathy with advanced renal failure: response to prednisone. AB - We report the case of a patient with acquired immunodeficiency syndrome (AIDS) who developed nephrotic syndrome and progressive renal failure mimicking human immunodeficiency virus (HIV)-associated focal segmental glomerulosclerosis (FSGS) who required initiation of hemodialysis and was found on renal biopsy to have membranous nephropathy. Hepatitis B and C serologies were negative. Although she required hemodialysis, she was treated with prednisone and experienced a progressive decline in her serum creatinine from 10.1 mg/dL to 1.9 mg/dL, which permitted the discontinuation of hemodialysis. After she abruptly discontinued prednisone, her creatinine level increased to 4.8 mg/dL, and she experienced marked worsening of her nephrotic syndrome. Resumption of prednisone resulted in normalization of serum creatinine and reduction in urine protein excretion. No adverse effects of prednisone occurred during this time. She remains off of hemodialysis for 1 year with a serum creatinine level of 1.0 mg/dL and urine protein excretion of 0.4 g/d. Although most patients with HIV infection, nephrotic-range proteinuria, and renal failure have FSGS, a minority may have membranous nephropathy. Although typically not a steroid-responsive lesion in the setting of advanced renal failure, membranous nephropathy may be a highly steroid responsive lesion in the HIV-infected patient, and treatment may help avert the need for dialysis in a patient population that generally has a poor outcome on dialysis. PMID- 9214411 TI - Combined gastric and ileocecal toxicity (serpiginous ulcers) after oral kayexalate in sorbital therapy. AB - Kayexalate (Roxane Labs, Columbus, OH) in sorbitol is commonly administered by the oral and rectal route for the treatment of hyperkalemia in patients with renal failure. It is believed to have minimal toxicity because it functions as a cation exchanger and is not absorbed. We herein report on a patient who developed identical serpiginous ulcers in the stomach and the terminal ileum after the use of Kayexalate. We believe that this drug could have significant gastrointestinal toxicity in specific patient groups and suggest tentative guidelines, both for the identification of such patients and for the safer use of Kayexalate in these circumstances. PMID- 9214412 TI - Thrombotic microangiopathy associated with alpha-interferon therapy for chronic myelocytic leukemia. AB - A 31-year-old man diagnosed as having chronic myelocytic leukemia (CML) developed renal insufficiency with nephrotic-range proteinuria during alpha-interferon (IFN) therapy for CML. A renal biopsy specimen showed remarkable thrombotic microangiopathic lesions resembling those of hemolytic-uremic syndrome. The patient had papules on both lower legs, and a cutaneous biopsy showed similar microangiopathic lesions in dermal and subcutaneous vessels. Although discontinuation of IFN and initiation of prednisolone therapy resulted in resolution of proteinuria, renal insufficiency persisted. These findings suggest that long-term IFN therapy can induce late-onset thrombotic microangiopathy in systemic microvessels. PMID- 9214413 TI - Enterovesical fistula presenting as life-threatening normal anion gap metabolic acidosis. AB - Enterovesical fistula is a rare complication of a variety of inflammatory and neoplastic diseases. It usually presents with pneumaturia, fecaluria, urinary tract infections, or irritable bladder symptoms in the setting of either diverticulitis or malignancy. For the first time, we describe a patient with an enterovesical fistula who presented with a life-threatening normal anion gap metabolic acidosis. The direction of flow through the fistula, ie, bladder to intestine, was contingent on a spastic bladder and was responsible for the atypical presentation. PMID- 9214414 TI - Propylene glycol-induced proximal renal tubular cell injury. AB - Propylene glycol is a solvent that is used in many oral, injectable, and topical medications. Although uncommon, acute renal failure has been attributed to propylene glycol. The mechanism of propylene glycol-mediated renal injury is unknown. We report a case of acute renal failure in a 16-year-old boy given large doses of pentobarbital and phenobarbital, both of which are solubilized with propylene glycol. A renal biopsy showed proximal renal tubular cell swelling and vacuole formation. The data from this case suggest that the reversible acute renal failure caused by propylene glycol is attributable to proximal renal tubular cell injury. PMID- 9214415 TI - Assessing health and quality of life outcomes in dialysis: a report on an Institute of Medicine workshop. PMID- 9214416 TI - Prednisone, renal function, and proteinuria in immunodeficiency virus-associated nephropathy. PMID- 9214417 TI - Immunotactoid glomerulopathy: report of a case. AB - We report a 51-year-old man diagnosed as having immunotactoid glomerulopathy (IT) who achieved partial remission after approximately 1 year of a low-dose prednisolone regimen. On admission, he was noted to show proteinuria, hypoproteinemia, and hypocomplementemia. On electron microscopy of the renal biopsy specimen, the mesangial and subendothelial areas were expanded because of the electron-dense deposits, which were represented by mostly straight and nonbranching hollow microtubule structures. The microtubular width was on average 22.0 nm. Clinical and histological findings did not support the diagnosis of amyloidosis, cryoglobulinemic glomerulonephritis, systemic lupus erythematosus, or paraproteinemias. Under treatment with oral prednisolone, 4 months later, the patient's serum albumin level increased from its lowest level of 2.3 to 3.6 g/dL, and CH50 from the lowest level of less than 6.3 to 32.4 U/mL. A 24-hour collection of urine showed that the protein had decreased from its highest level of 3.9 g to 2.0 g. This case suggests the effectiveness of long-term, low-dose prednisolone therapy for IT. PMID- 9214418 TI - Vasculitis in the central nervous system. PMID- 9214419 TI - Preliminary definition of improvement in juvenile arthritis. AB - OBJECTIVE: To identify a core set of outcome variables for the assessment of children with juvenile arthritis (JA), to use the core set to develop a definition of improvement to determine whether individual patients demonstrate clinically important improvement, and to promote this definition as a single efficacy measure in JA clinical trials. METHODS: A core set of outcome variables was established using a combination of statistical and consensus formation techniques. Variables in the core set consisted of 1) physician global assessment of disease activity; 2) parent/patient assessment of overall well-being; 3) functional ability; 4) number of joints with active arthritis; 5) number of joints with limited range of motion; and 6) erythrocyte sedimentation rate. To establish a definition of improvement using this core set, 21 pediatric rheumatologists from 14 countries met, and, using consensus formation techniques, scored each of 72 patient profiles as improved or not improved. Using the physicians' consensus as the gold standard, the chi-square, sensitivity, and specificity were calculated for each of 240 possible definitions of improvement. Definitions with sensitivity or specificity of <80% were eliminated. The ability of the remaining definitions to discriminate between the effects of active agent and those of placebo, using actual trial data, was then observed. Each definition was also ranked for face validity, and the sum of the ranks was then multiplied by the kappa statistic. RESULTS: The definition of improvement with the highest final score was as follows: at least 30% improvement from baseline in 3 of any 6 variables in the core set, with no more than 1 of the remaining variables worsening by >30%. The second highest scoring definition was closely related to the first; the third highest was similar to the Paulus criteria used in adult rheumatoid arthritis trials, except with different variables. This indicates convergent validity of the process used. CONCLUSION: We propose a definition of improvement for JA. Use of a uniform definition will help standardize the conduct and reporting of clinical trials, and should help practitioners decide if a child with JA has responded adequately to therapy. We are in the process of prospectively validating this definition and several others that scored highly. PMID- 9214420 TI - T cell responses to human type II collagen in patients with rheumatoid arthritis and healthy controls. AB - OBJECTIVE: To study the prevalence of T cell responses to human type II collagen (CII) in patients with rheumatoid arthritis (RA) with or without antibodies to CII, and in healthy controls. METHODS: Assays were performed to study T cell proliferative responses to CII in peripheral blood from 69 patients with RA (11 with anti-CII antibodies and 58 without) and 28 healthy controls. Further analysis was made of the time course of the response and the epitopic specificity, using peptides derived from the cyanogen bromide 11 (CB11) fragment of CII. RESULTS: Significant proliferative responses to CII were found in 50% of patients with anti-CII, 5.3% of RA patients without these antibodies, and 35.7% of healthy controls. Responses in RA patients differed from those in healthy controls; the former had kinetics suggestive of a recall response and the latter that of a primary response. Some common epitopes within CB11 were recognized by T cells from patients and controls. CONCLUSION: Proliferative T cell responses to CII occur in some healthy individuals, suggesting that thymic tolerance for this antigen may be incomplete. Most patients with RA have no evidence of a T cell response to CII, possibly indicating the development of peripheral tolerance to this antigen as a consequence of cartilage breakdown. However, in a minority of patients, T and B cell responses to CII persist, and may contribute to joint damage. PMID- 9214422 TI - Fibrosing alveolitis in systemic sclerosis: indices of lung function in relation to extent of disease on computed tomography. AB - OBJECTIVE: Thin-section computed tomography (CT) provides a sensitive and reproducible method of quantifying the morphologic extent of disease in the clinical management of fibrosing alveolitis associated with systemic sclerosis (FASSc). The aim of this study was to determine which indices of lung function best reflect the extent of disease on CT in FASSc, and to determine the independent influences of smoking history, extent of fibrosing alveolitis, demographic features, and concurrent treatment upon functional impairment in FASSc. METHODS: Sixty-four patients with FASSc were studied using CT and static and exercise lung function testing. Statistical relationships were determined by multiple regression analyses. RESULTS: Five patients with overt pulmonary hypertension were characterized by severe impairment in 3 indices of lung function: diffusing capacity for carbon monoxide (DLCO), DLCO adjusted for alveolar volume (KCO), and arterial partial pressure of oxygen. On multiple regression analysis, the major determinant of functional impairment was the extent of fibrosing alveolitis on CT. A history of smoking was independently associated with preservation of total lung capacity and depression of KCO, but did not otherwise influence functional-morphologic correlations. The percent predicted DLCO correlated better with extent of disease on CT (r = -0.70) than did oxygen desaturation on exercise (r = 0.55), the physiologic component of the clinical-radiographic-physiologic score (CRP index) (r = 0.52), or other indices of lung function. Lung volume measures correlated poorly with disease extent on CT. CONCLUSION: The percent predicted DLCO best reflects the extent of fibrosing alveolitis in FASSc, and therefore should be measured in routine evaluations. Exercise testing may also have a useful role in staging the severity of pulmonary fibrosis, but the CRP index offers no additional advantage over the DLCO and exercise testing. PMID- 9214423 TI - Scleroderma lung fibroblasts exhibit elevated and dysregulated type I collagen biosynthesis. AB - OBJECTIVE: To examine whether scleroderma lung fibroblasts show a pattern of aberrant type I collagen (CI) biosynthesis similar to that observed previously in studies of dermal fibroblasts in this disease. METHODS: CI secretion and steady state pro alpha1(I) collagen messenger RNA (mRNA) levels and COL1A2 gene activation were examined in fibroblasts grown from lung biopsy specimens obtained from 16 scleroderma patients with lung fibrosis and from 10 histologically normal lung specimens (controls). The effect of culture in a 3-dimensional (3-D) CI gel matrix culture on CI mRNA levels was also examined. RESULTS: The mean (+/- SEM) collagen secretion in monolayer culture for scleroderma lung fibroblasts was 90.9 +/- 56 ng/ml/10(6) cells, significantly greater (P < 0.05) than controls (40.2 +/ 17.5). Pro alpha1(I) collagen mRNA levels in monolayer cultures were higher in scleroderma (mean +/- SEM collagen:GAPDH ratio 3.7 +/- 0.9) compared with control (1.9 +/- 0.8) lung fibroblasts. Transient expression assays confirmed that genes coding for CI are transcriptionally activated in scleroderma lung fibroblasts compared with control strains. Although all lung fibroblasts induced equivalent contraction of 3-D CI gel matrices, scleroderma strains failed to show a reduction in steady-state pro alpha1(I) collagen mRNA levels in gel culture. CONCLUSION: We have demonstrated elevated CI biosynthesis and impaired mRNA down regulation for CI by scleroderma lung fibroblasts. These properties are likely to be highly relevant to the pathogenesis of scleroderma-associated lung fibrosis. PMID- 9214421 TI - Mycoplasma infection and rheumatoid arthritis: analysis of their relationship using immunoblotting and an ultrasensitive polymerase chain reaction detection method. AB - OBJECTIVE: To examine the relationship between infection with Mycoplasma and the development of rheumatoid arthritis (RA) and juvenile rheumatoid arthritis (JRA). METHODS: Immunoblotting of patient synovial fluid and sera on detergent-phase membrane protein extracts of various Mycoplasma species was carried out to learn whether patients exhibited serologic evidence of previous exposure to mycoplasmas. Moreover, an ultrasensitive polymerase chain reaction (PCR) method was developed for assessing whether Mycoplasma DNA could be detected in synovial fluid from patients and controls. RESULTS: Immunoblotting provided serologic evidence of previous Mycoplasma exposure in patients and controls. The genus specific PCR detected known human Mycoplasma species and could reliably detect <5 copies of Mycoplasma hominis, Mycoplasma fermentans, or a molecular mimic control in synovial fluid. Repeat testing revealed no evidence of Mycoplasma DNA in patient synovial samples. CONCLUSION: This study provided serologic evidence suggesting that, while previous exposure to Mycoplasma was common, there was no detectable persistence of Mycoplasma DNA in the synovial fluid or tissue of patients with RA or JRA. PMID- 9214424 TI - Pregnancy in relapsing polychondritis: twenty-five pregnancies in eleven patients. AB - OBJECTIVE: To determine maternal and fetal outcome in pregnant women with relapsing polychondritis (RP). METHODS: Retrospective review of 11 records selected from among those of 116 female RP (Michet's criteria) patients. RESULTS: In these 11 women, 25 pregnancies occurred after or concomitantly with the onset of RP. The mean (+/- SD) age at RP onset was 25 +/- 5 years (range 15-31). Therapeutic abortion was performed in 1 woman because of ongoing cyclophosphamide treatment. Disease flare occurred in 7 of 24 pregnancies, requiring treatment modification in 2 cases. RP was considered stable in 16 and asymptomatic in 1 of the 24. Treatment of RP was with nonsteroidal antiinflammatory drugs (2 of 24), steroids (13 of 24), dapsone (7 of 24), or plasma exchanges (1 of 24); no treatment was given in 8 of the 24 pregnancies. In 14 pregnancies, the course was uneventful. Ten pregnancies were complicated by ectopic nidation (n = 3), spontaneous abortion (n = 3), premature birth (n = 3), and premature rupture of membranes (n = 1). Eighteen normal children were born. CONCLUSION: Pregnancy does not seem to modify the course of RP. Successful pregnancies may be achieved in women with RP. No RP was observed in the neonates. PMID- 9214425 TI - Usefulness of procalcitonin for differentiation between activity of systemic autoimmune disease (systemic lupus erythematosus/systemic antineutrophil cytoplasmic antibody-associated vasculitis) and invasive bacterial infection. AB - OBJECTIVE: To investigate whether the determination of serum procalcitonin (PCT) in systemic autoimmune disease will help to discriminate invasive infection from highly active underlying disease. METHODS: Three hundred ninety-seven serum samples, from 18 patients with systemic lupus erythematosus (SLE) and 35 patients with systemic antineutrophil cytoplasmic antibody-associated vasculitis (AAV), were analyzed. Clinical disease activity was assessed by the Systemic Lupus Activity Measure in SLE patients and by the Birmingham Vasculitis Activity Score in AAV patients. Procalcitonin concentrations were determined in parallel with concentrations of neopterin, interleukin-6 (IL-6), and C-reactive protein (CRP). Additionally, serum creatinine values were obtained. RESULTS: In 321 of the 324 samples from the 42 patients with autoimmune disease but without systemic infection, serum PCT levels were within the normal range (i.e., <0.5 ng/ml), whereas the values for neopterin, IL-6, and CRP were elevated in patients with active underlying disease. All 16 systemic infections occurred in 11 patients with AAV, and were associated with PCT levels that were markedly elevated, to a mean +/- SD of 1.93 +/- 1.19 ng/ml. No correlation between the degree of renal impairment and PCT concentrations was seen. CONCLUSION: PCT may serve as a useful marker for the detection of systemic bacterial infection in patients with systemic autoimmune disease. PMID- 9214426 TI - Clinical, serologic, and immunogenetic features in Polish patients with idiopathic inflammatory myopathies. AB - OBJECTIVE: To determine the clinical, serologic, and immunogenetic correlations in patients with idiopathic inflammatory myopathies (IIM), and to evaluate the useful grouping of some diseases for practical clinical purposes. METHODS: Patients with IIM were categorized according to clinical presentation as compared with autoantibody specificity. Serum samples from 84 patients were screened for myositis-specific autoantibodies (MSAs) by indirect immunofluorescence and double immunodiffusion. All sera were also studied by protein A-assisted immunoprecipitation. Genomic DNA was isolated from peripheral blood mononuclear cells, and HLA-DQA1 and DRB1 alleles were determined. The patients were seen and followed up for many years in the same center. RESULTS: MSAs were present in 19% of patients. The most common MSAs were antisynthetases in 13% of patients (Jo-1 10.7%, PL-12 1.2%, and EJ 1.2%), associated with the antisynthetase syndrome. Anti-SRP was found in 1.2% of patients, associated with polymyositis, and anti-Mi 2 in 4.9%, found exclusively in patients with dermatomyositis. The most frequent MSA was PM-Scl in 23.8% of patients, associated with scleromyositis, and Ku was present in 9.6% of patients with overlap syndromes. The alleles that were found at a significantly increased frequency were HLA-DRB1*0301 (59.4%) and DQA1*0501 (71.6%), which are in linkage disequilibrium. DQA1*0501 was present in 85.7% of patients with antisynthetases, and in 100% of patients with PM-Scl and Ku. CONCLUSION: The HLA-DRB1*0301; DQA1*0501 haplotype was found to be significantly increased in this population overall and in those myositis patients with antisynthetase, anti-PM-Scl, and anti-Ku antibodies. The results of this study confirm that IIM are heterogeneous syndromes, but can be divided into more useful groups on the basis of clinical, serologic, and immunogenetic features. PMID- 9214427 TI - The AIMS2-SF: a short form of the Arthritis Impact Measurement Scales 2. French Quality of Life in Rheumatology Group. AB - OBJECTIVE: To develop a short form of the Arthritis Impact Measurement Scales 2 (AIMS2) questionnaire, preserving content validity as the priority criterion. METHODS: A 2-step reduction procedure was used: 1) Delphi technique, with 1 panel of patients and 1 panel of experts each selecting 1 set of items independently; and 2) nominal group technique, where members of both panels reached consensus on the final selection of items, using information derived from item analysis. Psychometric properties of the AIMS2-Short Form (AIMS2-SF) and AIMS2 were compared using data from a cohort of 127 rheumatoid arthritis patients who completed the AIMS2 twice prior to the initiation of methotrexate (MTX) treatment and 3 months post-initiation of MTX treatment. RESULTS: The 2 panels reached consensus on a 26-item AIMS2-SF (54.4% reduction from the AIMS2). Factor analysis showed preservation of the 5-component structure. Convergent validity (Physical and Symptom components with clinical variables: r = 0.24-0.59), test-retest reproducibility (intraclass correlation coefficient >0.7), and sensitivity to change at 3 months (standardized response mean 0.36-0.8, except Social Interaction component [0.08]) were very close to the values for the original AIMS2. CONCLUSION: The AIMS2-SF is a shorter version of the AIMS2 (i.e., available in 2-page format) and has psychometric properties similar to those of the AIMS2. PMID- 9214428 TI - Differential effects of aging on human chondrocyte responses to transforming growth factor beta: increased pyrophosphate production and decreased cell proliferation. AB - OBJECTIVE: To address the influence of age on inorganic pyrophosphate (PPi) accumulation in human articular chondrocytes. METHODS: Articular cartilage was obtained from men and women in 2 different age groups: ages 15-55 and 56-91. The effects of transforming growth factor beta1 (TGFbeta1) on PPi levels in the media and cell lysates of chondrocytes were investigated. In addition, the effects of TGFbeta on PPi accumulation were compared with chondrocyte proliferation. RESULTS: TGFbeta1 increased PPi levels to a greater extent in chondrocytes from subjects in the older age group compared with those obtained from younger subjects. Treatment of chondrocytes with TGFbeta1 led to a similar increase in total intracellular protein in both age groups. Although TGFbeta increased nucleoside triphosphate pyrophosphohydrolase activity and decreased alkaline phosphatase activity, these effects did not differ between the 2 age groups. Analysis of the same cell preparations showed an age-related decrease in TGFbeta induced chondrocyte proliferation, whereas these same cells showed an increased response with respect to PPi elaboration. CONCLUSION: These results show that aging differentially affected TGFbeta-induced PPi accumulation versus proliferation in human articular chondrocytes. These differences in TGFbeta response are likely to contribute to the development of age-associated cartilage diseases such as osteoarthritis. PMID- 9214429 TI - Alteration of cartilage metabolism by cells from osteoarthritic bone. AB - OBJECTIVE: To determine whether bone cells alter cartilage metabolism. METHODS: Bone cell cultures were established using explants obtained from the hip and knee joints of 9 patients with osteoarthritis (OA) and 6 subjects without arthritis (nonarthritic [NA]). NA human cartilage biopsy samples were incubated in the presence or absence of bone-derived cells, and the effects on glycosaminoglycan (GAG) release from cartilage were measured. RESULTS: Bone cell cultures secreted osteocalcin (OC) and did not contain cells expressing leukocyte common antigen. None of the 8 cultures established from NA bone, compared with 17 of 32 from OA bone, significantly altered GAG release from cartilage (P = 0.006). In knees with medial joint damage, 38% of the cultures derived from the medial side of the joint increased GAG release from cartilage. In contrast, 77% of the cultures derived from the lateral side of the joint had an effect on GAG, with 38% increasing and 38% decreasing GAG release. Seven cytokines were measured in OA bone cell supernatants. No significant difference was apparent in the concentration of any one cytokine when supernatants were compared according to their effects on GAG release. CONCLUSION: Bone cells from OA patients can influence cartilage metabolism. This might explain why increased subchondral bone activity can predict cartilage loss. PMID- 9214430 TI - Inhibition of rheumatoid synovial fibroblast proliferation by antisense oligonucleotides targeting proliferating cell nuclear antigen messenger RNA. AB - OBJECTIVE: To evaluate the feasibility of antisense oligonucleotides as therapeutic agents to inhibit synovial cell growth in rheumatoid arthritis (RA). METHODS: Fibroblast-like cells established from RA synovium were stimulated with interleukin-1beta (IL-1beta) and treated with antisense or sense oligonucleotides targeting proliferating cell nuclear antigen (PCNA) messenger RNA (mRNA). Proliferation of these cells was determined by 3H-thymidine incorporation. Effects of antisense oligonucleotides on the expression of mRNA and protein were evaluated by reverse transcriptase-polymerase chain reaction and immunohistochemical staining, respectively. RESULTS: Antisense oligonucleotides targeting PCNA inhibited IL-1-stimulated fibroblast proliferation, whereas sense oligonucleotides had no effect. Both mRNA and protein levels of PCNA were suppressed in the cells treated with antisense oligonucleotides, indicating that the antiproliferative effect was occurring through an antisense mechanism. CONCLUSION: These results suggest that antisense strategies designed to suppress PCNA expression have potential use as therapeutic agents for RA. PMID- 9214431 TI - Interleukin-1beta-stimulated invasion of articular cartilage by rheumatoid synovial fibroblasts is inhibited by antibodies to specific integrin receptors and by collagenase inhibitors. AB - OBJECTIVE: To study the role of integrin receptors in the invasion of cartilage by rheumatoid synovial fibroblasts (RSF). METHODS: RSF were cocultured with cartilage slices alone or in the presence of various potential activators or inhibitors. The penetration of the cartilage surface by RSF was determined by live-cell imaging of fluorescent-labeled cells. RESULTS: Interleukin-1beta (IL 1beta) and IL-8 stimulated the RSF invasion of cartilage. Invasion was specific for RSF and required a concentration gradient of IL-1beta. The IL-1beta-activated invasion of cartilage was inhibited by anti-IL-1 antibodies, IL-1 receptor antagonist, and collagenase inhibitors. RSF invasion was also inhibited by antibodies to alpha4, alpha5, alphaV, and beta1 integrins. CONCLUSION: In this study, an IL-1beta concentration gradient was required for RSF invasion into cartilage, raising the possibility that in vivo invasion may be induced by IL 1beta released by chondrocytes. The IL-1beta activation of RSF assayed in vitro may contribute to the RSF invasion of cartilage in vivo. Cartilage invasion requires the availability of beta1 and alpha4, alpha5, and alphaV integrins and the presence of collagenase activity. PMID- 9214433 TI - Use of T cell receptor/HLA-DRB1*04 molecular modeling to predict site-specific interactions for the DR shared epitope associated with rheumatoid arthritis. AB - OBJECTIVE: To use molecular modeling tools to analyze the potential structural basis for the genetic association of rheumatoid arthritis (RA) with the major histocompatibility complex (MHC) "shared epitope," a set of conserved amino acid residues in the third hypervariable region of the DRbeta chain. METHODS: Homology model building techniques were used to construct molecular models of the arthritis-associated DRB1*0404 molecule and a T cell receptor (TCR) from T cell clone EM025, which is specific for DR4 molecules containing the shared epitope sequence. Interactive graphics techniques were used to orient the TCR on the DR molecule, guided by surface complementarity analysis. RESULTS: The predicted TCR MHC-peptide complex involved multiple interactions and specificity for the shared epitope. TCR residues CDR1beta D30, CDR2beta N51, and CDR3beta Q97 were positioned to potentially participate in hydrogen bond interactions with the shared epitope DRbeta residues Q70 and R71. CONCLUSION: These results suggest a structural mechanism in which specific TCR recognition and possibly Vbeta selection are directly influenced by the disease-associated MHC polymorphisms. PMID- 9214432 TI - Adenosine A1 receptor promotion of multinucleated giant cell formation by human monocytes: a mechanism for methotrexate-induced nodulosis in rheumatoid arthritis. AB - OBJECTIVE: To determine why methotrexate (MTX) exacerbates rheumatoid nodules in some patients, despite the effective suppression of synovial inflammation. METHODS: Phorbol myristate acetate (PMA)-induced differentiation of monocytes into multinucleated giant cells was used as an in vitro model to study the effects of adenosine on nodulosis. RESULTS: MTX at 200-2,000 nM or the adenosine A1 agonist N5-cyclopentyl adenosine (CPA) (10(-12) to 10(-9) M) or the A2 antagonist 3,7-dimethyl-1-propargylxanthine markedly enhanced giant cell formation, whereas the adenosine A1 antagonist 8-cyclopentyl-dipropylxanthine completely reversed these effects. PMA, CPA, and MTX induced adenosine release by cultured monocytes at concentrations consistent with those associated with predominantly A1 effects. Furthermore, surface expression of A1 receptors was found to remain unchanged on the differentiating cells throughout the culture period. CONCLUSION: Agents that inhibit adenosine A1 receptors might be useful in the treatment of MTX-induced rheumatoid nodulosis, while still potentiating the A2-mediated antiinflammatory effects of MTX on synovitis. PMID- 9214434 TI - Involvement of endogenous kinins in the pathogenesis of peptidoglycan-induced arthritis in the Lewis rat. AB - OBJECTIVE: To investigate the pathophysiologic roles of endogenous bradykinin (BK) and des-Arg9-BK on local and systemic inflammatory responses in a rat model of acute arthritis induced by peptidoglycan-polysaccharide (PG-APS). METHODS: Female Lewis rats were injected intraperitoneally with PG-APS. Selective antagonists of B1 (Lys-[Leu8]-des-Arg9-BK) and B2 (Hoe 140) receptors were infused at 500 microg/kg and 5 mg/kg per day for 6 days, starting 3 days before induction of inflammation, with subcutaneous micro-osmotic pumps. The local inflammatory response was assessed by paw edema, joint swelling, and tissue content of BK and des-Arg9-BK. These peptides were measured by highly sensitive and specific chemiluminescent enzyme immunoassays. Systemic inflammatory reaction was evaluated by the hepatic concentration of the type 2 acute-phase protein T kininogen. RESULTS: PG-APS induced significant paw edema and joint swelling 24-72 hours after intraperitoneal injection. The maximal responses to PG-APS observed at 72 hours were significantly reduced (31-38%) by the combination of both B1 and B2 receptor antagonists at 5 mg/kg per day. PG-APS induced a significant increase of BK (up to 5.3-fold) and des-Arg9-BK (up to 4.1-fold) 72 hours after challenge. Liver T-kininogen content was increased by 5.3-, 7.7-, and 5.8-fold at 24, 48, and 72 hours, respectively, after PG-APS injection. At 24 hours, Hoe 140 and Lys [Leu8]-des-Arg9-BK increased liver T-kininogen content by 43% and 45%, respectively, but they had no effect at 72 hours. CONCLUSION: The results indicate that endogenous kinins are involved in local and systemic acute inflammatory responses, through both B1 and B2 kinin receptors, in the model of PG-APS-induced arthritis. PMID- 9214435 TI - Mechanisms of drug-induced lupus. III. Sex-specific differences in T cell homing may explain increased disease severity in female mice. AB - OBJECTIVE: To determine if sex-specific differences in lymphocyte trafficking could contribute to increased disease severity in female mice. METHODS: A lupus like disease was induced by injecting male and female mice with procainamide treated T cell clones. Trafficking was examined by labeling the injected cells with 51Cr or 5-chloromethylfluorescein diacetate. RESULTS: Females developed more autoimmune liver disease and greater titers of anti-DNA antibodies than did males, and 2-7 times more cells accumulated in the female spleens. Splenectomy prevented the development of autoantibodies and renal and liver disease. Oophorectomy decreased the splenic homing, autoantibody titer, and liver disease severity, to levels found in males. CONCLUSION: T cells traffic differently to the spleen in male and female mice, and the spleen appears to be essential in the disease process. This suggests that differences in T cell homing could contribute to sex-specific disease severity in this murine model, and also possibly in human disease. PMID- 9214437 TI - Clinical images: Skin hyperpigmentation associated with minocycline therapy. PMID- 9214436 TI - Human autoantibodies stabilize the quaternary structure of Ku antigen. AB - OBJECTIVE: To examine humoral immune responses to the native Ku antigen and to evaluate the role of autoantibodies in stabilizing intermolecular contacts between the p70 and p80 Ku subunits. METHODS: Recombinant free human p70 and p80 Ku subunits and p70/p80 heterodimers were expressed in Sf9 (insect) cells using baculovirus vectors. Affinity-purified recombinant human p70, p80, and p70/p80 dimer were studied by enzyme-linked immunosorbent assay (ELISA) and immunoprecipitation to evaluate autoantibody specificities in sera from 58 patients with systemic autoimmune disease. RESULTS: Anti-Ku antibodies were detected by ELISA or immunoprecipitation using K562 cell Ku antigen. All of the sera were reactive with the native recombinant p70, p80, or p70/p80 antigens: 47% were anti-p70+,anti-p80+ and 32% were anti-p70-,anti-p80+, but only 3% were anti p70+,anti-p80-. Unexpectedly, 18% of the sera recognized the p70/p80 dimer but did not recognize native p70 or p80 alone. A subset of sera containing autoantibodies that prevent the dissociation of p70 from p80 by high salt and detergent treatment was identified; monoclonal antibody (MAb) 162, a murine anti Ku MAb, displays the same property. Autoantibodies that stabilize the p70-p80 interaction were found most frequently in sera containing both anti-p70 and anti p80 antibodies. CONCLUSION: Autoantibodies to the native p80 subunit of Ku are more common than are anti-p70 antibodies. When anti-p70 antibodies were detected, they generally were found together with anti-p80. A novel type of autoantibody capable of stabilizing the p70/p80 heterodimer was identified in human sera for the first time. These "stabilizing" autoantibodies are found in sera containing both anti-p70 and anti-p80 antibodies, and also are produced by mice immunized with human Ku antigen. Autoimmunity to Ku may be initiated with an immune response to p80, followed by spreading to p70. We hypothesize that stabilizing antibodies could facilitate the spreading of autoimmunity from one subunit of Ku to another by altering the processing of p70 or p80 by antigen-presenting cells. PMID- 9214438 TI - Wegener's granulomatosis in pregnancy. AB - This report describes a case of severe limited Wegener's granulomatosis (WG) presenting in the third trimester of pregnancy with pansinusitis and necrotizing pneumonitis. The patient was treated successfully with a combination of corticosteroids and cyclophosphamide (CYC). The outcomes in the mother and the newborn were excellent. In a review of the English-language literature, we found 10 similar cases of WG with 13 pregnancies. WG occurring during pregnancy may have a more aggressive course and may require more aggressive treatment compared with WG occurring at other times. The treatment options for WG in pregnancy are discussed. PMID- 9214439 TI - Bone pain in a transplant patient. PMID- 9214440 TI - Correlation of serum IgG antibodies to recombinant P0 fusion protein with IgG antibodies to carboxyl-terminal 22 synthetic peptides and carboxyl-terminal 22 amino acid-deleted recombinant P0 fusion protein in patients with systemic lupus erythematosus. PMID- 9214441 TI - Antibodies to beta2-glycoprotein I in patients with systemic lupus erythematosus: comment on the articles by Tsutsumi et al and Roubey et al. PMID- 9214443 TI - Effects of interferon beta-1B in rheumatoid arthritis: a case report. PMID- 9214442 TI - Tumor necrosis factor a microsatellite polymorphism in rheumatoid arthritis: comment on the article by Hajeer et al. PMID- 9214444 TI - Measuring hepatitis C viremia in clinical samples: can we trust the assays? PMID- 9214445 TI - Resolution of paraneoplastic bile duct paucity following successful treatment of Hodgkin's disease. AB - We report the case of a 21-year-old woman who developed severe adult onset ductopenia in association with Hodgkin's lymphoma. Chemotherapy resulted in a remission of her Hodgkin's disease (HD) and significant improvement in liver function with resolution of the hepatic and biliary duct histological abnormalities, a therapeutic success not previously described in the literature. PMID- 9214446 TI - Cytotoxicity of bile salts against biliary epithelium: a study in isolated bile ductule fragments and isolated perfused rat liver. AB - We evaluated cytotoxic effects of different unconjugated and glycine- and taurine conjugated bile salts (BS) against bile duct epithelial cells in isolated bile ductule fragments and isolated perfused rat liver. Ultrastructural morphometric studies were performed in polarized rat bile ductule fragments exposed in vitro to increasing concentrations (10-100 micromol/L) of lithocholate (LCA), deoxycholate (DCA), chenodeoxycholate (CDCA), cholate (CA), ursodeoxycholate (UDCA), their taurine-conjugates, and glycoconjugates of cholic (GCA) or chenodeoxycholic acid (GCDCA) for 20, 30, or 75 minutes. To evaluate the cytotoxicity of unconjugated hydrophobic bile salts against biliary epithelium (BDE) in the whole liver, livers were isolated from rats with impaired taurine conjugation capacity (beta-alanine treatment) and perfused for 70 minutes with 2 micromol/min LCA (n = 6), CDCA (n = 6), CA (n = 6), or 0.5 micromol/min tauro-LCA (n = 4). In isolated bile ductule fragments, hydrophobic unconjugated bile salts (LCA, CDCA, DCA) induced a marked damage of intracellular organelles, mainly mitochondria. The damage started at a concentration of 10 micromol/L and became prominent at concentrations higher than 50 micromol/L. No damage of the apical and basolateral membrane was seen and tight junctions appeared intact. UDCA, taurine and glycoconjugated bile salts failed to induce any evident ultrastructural alteration. In taurine-depleted isolated livers, perfused with LCA, CDCA, or CA, bile duct epithelial cells showed no evidence of intracellular damage, despite the increased biliary excretion of unconjugated BS. Marked alterations of the apical cell membrane were seen only in livers perfused with LCA and in isolated segments of the biliary epithelium. In contrast with biliary epithelium, hepatocytes showed prominent subcellular damage with CA and CDCA, and profound alterations of the canalicular membrane with LCA and tauro-LCA. We have shown that, in vitro, BDE cells are not damaged by taurine- or glycine-conjugated BS, but they are very sensitive to cytotoxicity of hydrophobic unconjugated BS. Such sensitivity is not present in the whole liver, probably because of the specificity of BS transport processes, the microvascular architecture of the bile ductal system, and the presence in bile of a physiological surfactant, such as phospholipids. PMID- 9214447 TI - Does antimitochondrial antibody status affect response to treatment in patients with primary biliary cirrhosis? Outcomes of ursodeoxycholic acid therapy and liver transplantation. AB - Approximately 5% to 10% of patients with features otherwise consistent with primary biliary cirrhosis (PBC) lack antimitochondrial antibodies (AMA). Most of these patients have other autoantibodies, a syndrome recently named "autoimmune cholangitis." We report our experience in patients with AMA-negative PBC treated with ursodeoxycholic acid (UDCA) and/or liver transplantation (OLT). The study of response to UDCA was performed as follows. While recruiting patients for a previously reported multicenter trial, we identified 8 patients with AMA-negative PBC. The patients were given UDCA and followed up at regular intervals. The characteristics of AMA-negative patients at presentation were similar to those of AMA-positive patients with PBC. The clinical outcomes and sequential liver biochemistries of UDCA treatment were also comparable with those of AMA-positive patients. The study of outcome of OLT was performed as follows. We identified OLT recipients at the Mayo Clinic who had clinical, radiological, and histological features compatible with PBC. Their pretransplant AMA status was determined, and each AMA-negative patient was paired with 2 AMA-positive patients. Of 85 OLT recipients with a diagnosis of PBC, 6 (7.1%) were AMA negative, including 1 who had undergone UDCA therapy. After a median of 36 months of follow-up, graft and patient survival rates and subsequent histological changes (disease recurrence and steroid-resistant or late rejections) were comparable in AMA-negative and positive PBC patients. In summary, in our experience of 13 AMA-negative PBC patients (including 9 who met the criteria for a diagnosis of autoimmune cholangitis), treatment with UDCA or OLT resulted in similar outcomes to those found in AMA-positive patients. We conclude that AMA status does not affect the response in PBC patients to treatment with UDCA or OLT. PMID- 9214448 TI - Enhanced G-protein-induced relaxation in portal hypertensive rats: role of nitric oxide. AB - Portal hypertension (PHT) is characterized by splanchnic hyperemia due to a reduction in mesenteric vascular resistance. The reasons for the decreased resistance include an increased responsiveness to a vasodilator substance. Because the activation of an inhibitory guanine nucleotide regulatory (Gi) protein can result in endothelium-dependent relaxation, we tested the hypothesis that exaggerated Gi-protein induced relaxation via a nitric oxide (NO)-dependent pathway partly reflects the enhanced Gi-protein expression in PHT vessels. PHT was created in Sprague-Dawley rats by a partial portal-vein ligation. Control animals were sham operated. Using isolated vascular rings in the absence or presence of an intact endothelium, N(G)-nitro-L-arginine methyl ester (L-NAME), and pertussis toxin, dose response relationships for sodium fluoride (NaF; range, 0.1-4 mmol/L), a Gi protein activator, were determined in a cumulative manner. Gi protein expression was determined by Western blotting. NaF caused a dose dependent relaxation in both sham and portal hypertensive pre-contracted vessels, an effect that was significantly inhibited by pertussis toxin, endothelial denudation, and L-NAME. Concentrations of NaF greater than 4 mmol/L caused contractions, an effect that was unaffected by L-NAME. The NaF-induced relaxation response was significantly greater in PHT vessels as compared with sham concomitant with increased Gi-protein expression in PHT vessels. These data suggest that the enhanced endothelial Gi-protein-induced relaxation in PHT vessels may partly reflect enhanced expression of Gi-proteins in PHT vessels and may, thus, represent an important mechanism for exaggerated NO-dependent relaxation in the PHT vasculature. PMID- 9214449 TI - Hemodynamic evaluation of the addition of isosorbide-5-mononitrate to nadolol in cirrhotic patients with insufficient response to the beta-blocker alone. AB - The association beta-blockers plus isosorbide-5-mononitrate (I5M) has been proposed for the treatment of portal hypertension in patients with insufficient response to beta-blockers alone, according to hemodynamic criteria. The mechanism of action in these patients is not clearly defined. Fifteen patients with cirrhosis and esophageal varices were evaluated by hepatic venous pressure gradient (HVPG) measurement and duplex-Doppler ultrasonography before and after 1 month of treatment with nadolol. Nine patients who did not exhibit a decrease in HVPG to 12 mm Hg or a percent decrease greater than 20% were classified as poor responders, and were studied again with the same methodology after 3 months of chronic administration of nadolol + I5M 20 mg twice per day. In poor responders, mean HVPG decrease after nadolol was 8.9% +/- 2.8%, and after the combination, it was 25.7% +/- 1.7% (P = .004). All patients except one became good responders to the association. Portal blood flow (PBF) decreased significantly after nadolol (P = .004), and remained unchanged after the addition of nitrates. Resistance to portal blood flow (RPBF) increased after nadolol (P = .02) and returned to baseline values during combined treatment (P = .03). In good responders, an adequate decrease in HVPG was associated with a decrease in PBF (P = .06) but no change in RPBF. A wide spectrum of combined changes in PBF and in RPBF after nadolol was observed in poor responders, ranging from no change in either parameter to a marked decrease in PBF counterbalanced by a marked increase in RPBF. The addition of I5M was followed in most cases by larger effects on resistance than on flow. Doppler parameters were not significantly correlated with the HVPG response to nadolol alone or associated with I5M. It is concluded that good hemodynamic responders to nadolol differ from poor responders in the lack of increase in RPBF after the drug. The addition of nitrates to nadolol is effective in decreasing portal pressure in most poor responders to nadolol alone. A decrease in outflow resistance is the main mechanism involved. PMID- 9214450 TI - Taste perception in cirrhosis: its relationship to circulating micronutrients and food preferences. AB - Impairment of gustatory acuity may influence nutrient intake and hence nutritional status. The aim of this study was to evaluate gustatory acuity in patients with cirrhosis and its relationship to circulating concentrations of micronutrients, and food preferences. Gustatory evaluation was undertaken, using a rinsing technique, in 75 cirrhotic patients and 75 comparable healthy volunteers. Circulating concentrations of magnesium, zinc, vitamin A, and alpha- and beta-carotene were measured, and food preferences were assessed by questionnaire. The cirrhotic patients showed impaired gustatory function with significantly higher (less sensitive) median thresholds for detection of salt, sweet, and sour and for recognition of bitter, salt, sweet, and sour, together with a higher overall median gustatory score (P < .0001). Mean circulating concentrations of magnesium, zinc, vitamin A, and alpha- and beta-carotene were significantly lower in the patient population. Serum magnesium was significantly negatively associated with detection of salt (P = .02) and gustatory score (P = .02). Patients' subjective assessment of taste acuity did not correspond with objective measurements. Overall, no differences were observed in food preferences between the two groups, nor was any association found between food preferences and gustatory acuity. Patients with cirrhosis have impaired gustatory acuity that is associated with hypomagnesemia but apparently does not affect food selection. PMID- 9214451 TI - Nonparenchymal cells from regenerating rat liver generate interleukin-1alpha and 1beta: a mechanism of negative regulation of hepatocyte proliferation. AB - Following experimental partial hepatectomy of 70% in the rat, there is a semisynchronized surge of hepatocyte proliferation that ceases after 48 to 72 hours. Little is known about the determinants governing the termination of the proliferative phase, although transforming growth factor (TGF) beta has been implicated as an important inhibitor of hepatocyte replication in this model. We previously reported an additional non-TGF-beta inhibitor in medium conditioned by nonparenchymal cells isolated from regenerating liver (CM-NPC-Reg) between 24 and 48 hours after partial hepatectomy, but it was not found in medium conditioned by nonparenchymal cells from unoperated control liver. CM-NPC-Reg suppressed replicative DNA synthesis of primary rat hepatocytes in response to hepatocyte growth factor (HGF), epidermal growth factor (EGF), or TGF-alpha as assessed by 3H-thymidine incorporation. We now present evidence that interleukin (IL)-1 is the major inhibitor of hepatocyte DNA synthesis present in CM-NPC-Reg. IL-1 receptor antagonist abrogated the inhibition, as did antibodies to rat IL-1alpha and -beta; a combination of both antibodies was required, implicating both IL 1alpha and IL-1beta as active constituents in CM-NPC-Reg. To investigate in vivo changes in IL-1 expression, we assessed expression of IL-1alpha messenger RNA (mRNA) in whole rat liver following partial hepatectomy; mRNA was down-regulated at 10 hours in the pre-replicative phase of liver regeneration and up-regulated at 24 hours and 48 hours when proliferation is waning. Rat hepatocytes isolated from liver 24 hours after partial hepatectomy showed increased sensitivity to the inhibitory action of IL-1. Exogenous IL-1beta, administered parenterally to a group of rats at 0 and 12 hours after partial hepatectomy significantly reduced the incorporation of the thymidine analogue, bromodeoxyuridine (BrdU), into hepatocytes at 18 hours. These data indicate that nonparenchymal cells isolated from regenerating rat liver elaborate IL-1, and support the hypothesis that IL-1 plays a role suppressing hepatocyte proliferation and terminating the surge of DNA synthesis induced after partial hepatectomy. PMID- 9214453 TI - Hepatic tocopherol content in primary hepatocellular carcinoma and liver metastases. AB - The high incidence of hepatocellular carcinoma (HCC) in cirrhosis, where previous studies have indicated a severe reduction in several antioxidant vitamin factors, prompted us to compare plasma liposoluble vitamins with tocopherol content in healthy and neoplastic liver tissue in humans. This, with a view to a more positive preventive dietary approach, given the conflicting results obtained by liposoluble vitamin dietary supplementation in different malignancies. Eleven patients with cirrhosis, 18 patients affected by cirrhosis with HCC, and 10 patients with liver metastases (LM) from digestive tract adenocarcinomas were compared with controls who had undergone perlaparoscopic cholecistectomy. Plasma alpha- and beta-carotene, retinol and tocopherol, together with liver tocopherol, from both nonmalignant portions and malignant nodules of the same organ, were determined by high-performance liquid chromatography following a well-assessed technique. The results confirm a trend towards a reduction in circulating carotenoids and tocopherol in cirrhosis and in patients affected by cirrhosis with HCC. Tocopherol content in liver tissue is significantly decreased in cirrhosis (0.26 + 0.03 micromol/g prot., mean + SEM, P < .001) and in cirrhotic areas of the HCC group (0.31 + 0.02, P < .002), with respect to its content in liver specimens of healthy controls (0.46 + 0.03) and in healthy areas of the same organ in patients with LM (0.41 + 0.03). Tocopherol concentration is further reduced by 50% in malignant liver nodules of HCC, with respect to surrounding cirrhotic tissue, whereas in metastatic liver nodules from digestive neoplasms the tocopherol content is almost twice that of healthy surrounding areas. This unpredictable tocopherol behavior in liver specimens, of secondary as opposed to primary malignancies of the liver, affords further insight into the conflicting effects of liposoluble vitamins employed in the chemopreventive treatment of different malignant diseases, where hepatic tocopherol concentration show opposite trends: halved in primary HCC and doubled in LM of digestive adenocarcinomas, with respect to healthy controls. PMID- 9214452 TI - Co-expression and regulation of Met and Ron proto-oncogenes in human hepatocellular carcinoma tissues and cell lines. AB - Met and ron proto-oncogenes encode the cell surface receptors for hepatocyte growth factor (HGF) and hepatocyte growth factor-like (HLP) protein, respectively, and induce mitogenesis, motogenesis, morphogenesis, and metastatic activity in various cell types. Overexpression of met in human carcinoma has been reported by several groups including ours; however, the mechanisms that control met gene expression are thus far unclear. The present study focuses on the expression and regulation of the Met and Ron receptors in human hepatocellular carcinoma (HCC). We report here that abnormal expression of met and ron proteins occurs in some cases of human HCC. Using several HCC cell lines as a model system, we show that HGF, as well as other cytokines, such as epidermal growth factor (EGF), interleukin-1 (IL-1), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-alpha), induce met and ron expression. Using several chimeric constructs consisting of various lengths of the met promoter region fused to the reporter gene of chloramphenicol acetyl transferase (CAT), and by performing transient transfection of these constructs into HepG2 cells, we show that induction of met gene expression by HGF and other cytokines is, at least in part, through up-regulation of met gene promoter activity. The DNA region conferring responsiveness to cytokine induction was located within 0.2 kb of the met core promoter. Interestingly, EGF did not stimulate met promoter activity in any of the met-CAT chimeric constructs. These results provide evidence that met and ron are modulated in the liver by a similar cytokine network. In the case of met expression, the 0.2-kb region in the met gene promoter may play an important role in mediating its gene induction in response to HGF and other cytokines. Our results also suggest that unregulated expression of met and ron may be associated with pathological conditions, such as HCC, in the liver. PMID- 9214454 TI - Mechanism of the impairment of the glucagon-stimulated phosphoenolpyruvate carboxykinase gene expression by interleukin-6 in rat hepatocytes: inhibition of the increase in cyclic 3',5' adenosine monophosphate and the downstream cyclic 3',5' adenosine monophosphate action. AB - In cultured rat hepatocytes, the gluconeogenic key enzyme, phosphoenolpyruvate carboxykinase (PCK), is induced by glucagon via elevation of cyclic 3',5' adenosine monophosphate (cAMP). The proinflammatory cytokine, interleukin-6 (IL 6), which in the liver together with IL-1beta and tumor necrosis factor alpha triggers the acute-phase response, had been shown to attenuate the glucagon induced increase in PCK gene transcription, messenger (mRNA) levels, and enzyme activity. The molecular mechanism of this inhibition was investigated in the present study. Glucagon increased cyclic cAMP and PCK mRNA levels to a transient maximum twofold and fivefold, respectively. The increases were attenuated by IL 6. Forskolin, which stimulates adenylate cyclase activity, increased cAMP and PCK mRNA levels 1.6-fold and fivefold, respectively. However, IL-6 attenuated the forskolin-stimulated increase in PCK mRNA but not the increase in cAMP. This showed that IL-6 inhibited PCK mRNA increase in part by the attenuation of cAMP increase, but also beyond cAMP formation. This was confirmed in experiments in which PCK mRNA levels were increased by the nonhydrolyzable cAMP-analogue, chlorophenylthio (CPT)-cAMP. The increase in PCK mRNA was again attenuated by IL 6. In pertussis toxin- and in isobutylmethylxanthine-treated hepatocytes, IL-6 still inhibited the glucagon-stimulated increase in cAMP, indicating that IL-6 did not activate an inhibitory G-protein or phosphodiesterase, which could cause the impairment of cAMP increase. To demonstrate whether the inhibition of PCK gene expression by IL-6 beyond cAMP might be caused by the inhibition of the activation of the PCK gene promoter by cAMP, cultured rat hepatocytes were transfected with a luciferase reporter gene construct under the control of a PCK gene promoter fragment (base -979 to base +32). Luciferase activity was determined after stimulation of the cells with CPT-cAMP in the absence or presence of IL-6. CPT-cAMP increased luciferase activity by 1.7-fold, which was inhibited in the presence of IL-6. It is concluded that IL-6 had a dual inhibitory effect on the stimulation of PCK gene expression by glucagon. It inhibited the increase in cAMP at a site before cAMP formation by adenylate cyclase and at a site after cAMP formation, the activation of the PCK gene promoter by cAMP. PMID- 9214455 TI - Preventive and therapeutic effects in rats of hepatocyte growth factor infusion on liver fibrosis/cirrhosis. AB - Liver fibrosis/cirrhosis is characterized by hyper-accumulation of fibrous tissue components and is commonly observed in later or terminal states of chronic hepatic diseases. In ongoing work, we found that the administration of human recombinant hepatocyte growth factor (hrHGF) suppressed the onset of liver fibrosis/cirrhosis in several distinct models and accelerated the recovery from liver fibrosis/cirrhosis in rats. Repeated administration of porcine serum for 10 weeks to rats induced liver fibrosis without any accompanying hepatocellular injuries; in addition, the intravenous (i.v.) administration of hepatocyte growth factor (HGF) to these rats suppressed increases in fibrous components and hydroxyproline contents in the liver, thus preventing the onset of liver fibrosis. Repeated administration of dimethylnitrosamine (DMN) for four weeks induced liver cirrhosis, as characterized by the hyper-accumulation of fibrous components, infiltration of mononuclear leukocytes, and hepatic dysfunction. When HGF was injected daily for four weeks along with DMN-treatment, the onset of DMN induced hepatic fibrosis/cirrhosis was suppressed; the numbers of infiltrating mononuclear cells, fibrous tissue components, and hydroxyproline content in the liver were decreased. When HGF was injected for two weeks following four weeks of DMN-treatment, HGF accelerated the recovery from liver cirrhosis and prevented death due to hepatic dysfunction. Likewise, HGF-injection suppressed the onset of liver fibrosis, when liver fibrosis had been induced by long-term treatment with carbon tetrachloride (CCl4). Thus, the administration of HGF holds great promise for treating subjects with liver fibrosis/cirrhosis as a result of chronic hepatic injury. PMID- 9214456 TI - Cholesterol supplementation prevents necrosis and inflammation but enhances fibrosis in alcoholic liver disease in the rat. AB - Based on studies that show a role for the low-density lipoprotein (LDL)-receptor in arachidonic acid delivery and eicosanoid synthesis in macrophages, the present study investigated the effect of cholesterol supplementation on pathological changes and thromboxane (TX) synthesis in alcoholic liver injury. Male Wistar rats were intragastrically fed ethanol with either corn oil or fish oil for 1 month. Control rats received isocaloric amounts of dextrose instead of ethanol. An additional group of rats fed either ethanol or dextrose with fish oil or corn oil were supplemented with 1% cholesterol. At the time of killing, all rats had the following evaluated: liver histopathology, lipid peroxidation, liver and plasma thromboxane levels, plasma endotoxin and messenger RNA (mRNA) levels of LDL-receptor, tumor necrosis factor alpha (TNF-alpha), cyclooxygenase (Cox)-1 and -2, and transforming growth factor beta (TGF-beta). Rats fed ethanol with either fish oil or corn oil developed fatty liver, necrosis, inflammation, and central vein collagen deposition. Cholesterol supplementation enhanced the degree of fibrosis but prevented necrosis and inflammation. These alterations in pathological changes by cholesterol were accompanied by absent TNF-alpha and Cox 2 mRNAs, decreased thromboxane levels, decreased lipid peroxidation, and increased TGF-beta mRNA. Cholesterol enrichment of the diet thus decreases proinflammatory components, but enhances fibrosis in ethanol-fed rats. PMID- 9214457 TI - C1Q synthesis by tissue mononuclear phagocytes from normal and from damaged rat liver: up-regulation by dexamethasone, down-regulation by interferon gamma, and lipopolysaccharide. AB - The subcomponent of complement C1, C1q, mediates complement activation via the classical pathway, and therefore may play an important role in the inflammatory processes in which complement activation is involved. The aim of our study was to investigate C1q synthesis by macrophages of normal and of acutely damaged livers. The localization of C1q in liver tissue was studied by immunohistochemistry. Rat liver tissue macrophages were isolated from normal as well as from acutely damaged (carbon tetrachloride model) liver, and were separated into small, monocyte-like phagocytes and large, mature tissue macrophages, as revealed by immunocytochemistry. C1q gene expression was studied by endogeneous labeling of newly synthesized proteins, immunoprecipitation, and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and by reverse-transcription polymerase chain reaction (RT-PCR) of C1qB messenger RNA (mRNA). Semiquantitative analysis was performed by Northern blotting of total RNA and hybridization with the radioactively labeled RT-PCR product. C1 esterase inhibitor synthesis was studied in parallel. For comparison, C1q and C1-inhibitor synthesis were also investigated in blood monocytes and peritoneal macrophages. C1q was weakly detectable in sinusoidal cells of the normal liver. C1qB mRNA, as well as constitutive synthesis and secretion of C1q, was clearly detected in freshly isolated and cultured Kupffer cells from normal rat liver. In comparison, newly recruited "inflammatory" macrophages from damaged rat liver synthesized considerably lower amounts of the protein, similar to what was found in the monocyte-like macrophages of normal liver and in peritoneal macrophages. Monocyte C1qB mRNA was not detected even by RT-PCR, and remained undetectable during the time in culture. Similar behavior was observed for C1-inhibitor synthesis. Treatment of the cultures with interferon gamma (IFN-gamma) or lipopolysaccharide (LPS) strongly decreased, whereas treatment with dexamethasone strongly increased C1q gene expression in the macrophage populations, and induced C1qB mRNA in cultured monocytes, as revealed by RT-PCR. Kupffer cells of normal liver may produce considerable amounts of C1q, whereas the inflammatory macrophages of the acutely damaged liver may not be so important for the synthesis of C1q. PMID- 9214458 TI - Glycine protects against hepatocyte killing by KCN or hypoxia by preventing intracellular Na+ overload in the rat. AB - Glycine has been shown to prevent hepatocyte death induced by anoxia and by several toxic agents. However, the mechanisms responsible for such a cytoprotective effect have not yet been entirely clarified. We have previously shown that an uncontrolled increase in intracellular Na+ is critical for hepatocyte killing induced by adenosine triphosphate (ATP) depletion. We herein report that protection by glycine (2 mmol/L) against cytotoxicity induced in isolated rat hepatocyte by potassium cyanide (KCN) or hypoxia was associated with the prevention of cytosolic Na+ accumulation. The addition of the Na+ ionophore, monensin, abolished the effects of glycine on both Na+ increase and cytotoxicity. Pretreating hepatocytes with the glycine-receptor antagonist, strychnine (1 mmol/L), similarly prevented Na+ overload and cell killing. Glycine at high concentrations and strychnine are known to block Cl- channels in many cell types. Consistently, we have observed that glycine and strychnine prevented the increase of intracellular Cl- levels caused by hypoxia or KCN. Incubation of hepatocytes in a Cl(-)-free medium, obtained by substituting chloride with membrane impermeable gluconate, significantly reduced Na+ accumulation and cell killing triggered by hypoxia or KCN. Both these effects were abolished by the addition of monensin. The cytoprotective action exerted by hepatocyte incubation in the Cl(-) free medium was, however, lost when membrane-permeable nitrate, which allowed Na+ accumulation, was used instead to replace chloride. Altogether, these results indicate that glycine inhibition of Cl- conductance protects against hepatocyte killing induced by KCN and hypoxia by interfering with intracellular Na+ accumulation triggered by ATP depletion. PMID- 9214459 TI - Tumor necrosis factor-induced, superoxide-mediated neutrophil accumulation in cold ischemic/reperfused rat liver. AB - The mechanisms of hepatic ischemia/reperfusion injury are complicated and multifactorial. This study was designed to examine superoxide generation and neutrophil accumulation in cold ischemic-reperfused rat livers after elimination of Kupffer cells and to determine the role of superoxide/tumor necrosis factor (TNF) interactions. Rat Kupffer cells were eliminated by liposome-encapsulated dichloromethylene diphosphonate injected intravenously. Livers from control and treated rats were isolated and preserved in University of Wisconsin solution (4 degrees C) for 0, 12, and 24 hours and then perfused for 60 minutes with oxygenated Krebs-Henseleit bicarbonate buffer (37 degrees C) by adding neutrophils into the perfusate. Superoxide generation was measured by using real time chemiluminescence (CL) during perfusion, and neutrophil accumulation was assessed by measuring myeloperoxidase activity in the liver tissue. In the control livers, CL intensity markedly increased on reoxygenation, and after neutrophil infusion it increased again with a lag period of 10 minutes. Total CL intensity and myeloperoxidase activity increased with the duration of cold preservation. TNF release into the effluent perfusate was detectable only after 24 hours of preservation, and lactate dehydrogenase release was high. Elimination of Kupffer cells attenuated CL intensity and TNF and lactate dehydrogenase release and resulted in reduced myeloperoxidase activity. Electron microscopy revealed amelioration of hepatocyte swelling and endothelial cell disruption when Kupffer cells were eliminated. After 24 hours of preservation, superoxide generation was inhibited in the control livers by anti-TNF antiserum, whereas TNF release was not inhibited by superoxide dismutase. These results suggest that TNF induces superoxide generation by Kupffer cells, which mediates neutrophil accumulation and causes cellular injury in the initial phase of reperfusion. PMID- 9214460 TI - Effects of S-adenosyl-L-methionine on hepatic and renal oxidative stress in an experimental model of acute biliary obstruction in rats. AB - We used an animal model of extrahepatic biliary obstruction of 7 days' duration to study the production of thiobarbituric acid reactive substances (TBARS), total glutathione (TG), reduced glutathione (GSH), and oxidized glutathione (GSSG), and the enzymatic activities of GSH-peroxidase, GSSG-reductase, and GSH-transferase. Four groups of six rats each were treated with saline, drug solvent, S-adenosyl-L methionine (SAM) 5 mg/kg/d, subcutaneously, or SAM 10 mg/kg/d, subcutaneously. Extrahepatic biliary obstruction increased TBARS. SAM had the dose-dependent effects of inhibiting TBARS production and increasing TG content, mainly as a result of the increase in GSH. The activity of GSH-peroxidase and GSH-transferase was also significantly increased. In renal tissue these effects were statistically significant only in animals given the higher dose of SAM. In liver we found a reduction in biochemical values indicative of liver damage. We conclude that effect of SAM on hepatorenal function is strongly influenced by the drug's ability to reestablish equilibrium after oxidative tissue stress. PMID- 9214461 TI - Severe radiation-induced liver disease following localized radiation therapy for biliopancreatic carcinoma: activation of hepatic stellate cells as an early event. AB - Radiation-induced liver disease is recorded as a form of veno-occlusive disease. Its pathogenesis remains unclear even if the initial injury likely occurs in the endothelial cells of central veins. The aim of our study was to investigate liver morphological features in relation to alpha-isoform of smooth muscle actin expression in hepatic stellate cells in six patients treated by localized radiotherapy on the biliopancreatic area. Within the month after completion of treatment, an activation of hepatic stellate cells strictly confined to irradiated areas and coinciding with congestive changes was observed. At a later stage, collagen deposition gradually increased, replacing the congestive and destroyed areas. This new fibrotic tissue also contained numerous alpha-smooth muscle positive cells. Our data suggest that early hepatic stellate cells activation coinciding with congestive changes plays an important role in radiation liver injury and ensuing fibrosis. PMID- 9214462 TI - In situ detection of lipid peroxidation by-products in chronic liver diseases. AB - Lipid peroxidation is an autocatalytic mechanism leading to oxidative destruction of cellular membranes. The deleterious consequences of this mechanism are related in part to the formation of reactive aldehydic products that bind to intra- or extracellular molecules to form adducts. Specific antibodies directed against malondialdehyde (MDA) and 4-hydroxynonenal (HNE) adducts, major aldehydic metabolites of lipid peroxidation, allowed us to investigate in situ, with an immunohistochemical procedure, the occurrence of lipid peroxidation in a panel of different chronic liver diseases. Intracellular HNE and MDA adducts were detected respectively in 24 of 39 cases (62%) and in 12 of 34 cases investigated (35%). They were localized mainly in the cytoplasm of hepatocytes, with the strongest staining observed in hemochromatosis, Wilson's disease, and in areas of acute alcoholic hepatitis in cases of alcoholic liver diseases. A peculiar pattern of immunostaining was observed in primary biliary cirrhosis where biliary cells of destroyed but also intact bile ducts strongly expressed HNE adducts. The liver extracellular matrix also displayed MDA adducts (30 of 34 cases, 88%) and HNE adducts (23 of 39 cases, 59%). While HNE adducts were specifically localized on large bundles of collagen fibers, MDA adducts were detected in a thin reticular network and in sinusoidal cells around portal tracts or fibrous septa. In conclusion, lipid peroxidation by-products are detectable in chronic liver diseases. Immunohistochemical results suggest that this mechanism is implicated very early in the pathogenesis of some of these diseases. PMID- 9214463 TI - Association of a tumor necrosis factor promoter polymorphism with susceptibility to alcoholic steatohepatitis. AB - Twin concordance studies suggest that genetic factors play a role in determining why only a minority of heavy drinkers develop hepatitis and cirrhosis. Tumor necrosis factor alpha (TNF-alpha) has emerged as the "final common pathway" in the pathogenesis of alcohol-related hepatic necro-inflammation. We have examined the frequency of the two recently described polymorphisms of the TNF-alpha promoter in 150 patients with biopsy-proven alcoholic liver disease and 145 healthy volunteers. There was a significant excess of the rare allele (TNFA-A; G( 238) --> A) at position -238 in patients with steatohepatitis compared with controls or patients without this lesion. This is consistent with previous suggestions that the TNFA-A allele, which falls within a putative Y regulation box of the TNF-alpha promoter, is associated with increased TNF-alpha expression. No differences were observed for the polymorphism at position -308. PMID- 9214464 TI - Nitric oxide formation lowers norepinephrine-induced intrahepatic resistance without major effects on the metabolism in the perfused rat liver. AB - Nitric oxide (NO) and norepinephrine are potent vasoactive agents that are involved in the control of portal blood flow. We have studied NO and norepinephrine in a non-recirculated rat liver perfusion to analyze their influence on portal flow and hepatic metabolism. Animals were either pretreated with endotoxin (4 hours; 10 mg/kg intraperitoneally) to activate the inducible NO synthase (NOS2), or used without pretreatment for the constitutive NO synthase (NOS3). In both groups, portal flow, bile flow, bile secretion, and the sinusoidal bile acid uptake were reduced by norepinephrine with a simultaneous increase of glucose and lactate output. The addition of the substrate for NO synthesis, L-arginine (0.5 mmol/L), to the perfusate markedly inhibited the effect of 0.1 micromol/L norepinephrine on portal flow from -2.6 +/- 0.32 to 0.3 +/- 0.1 mL/g/10 min in endotoxin-treated animals, and from -2.9 +/- 0.45 to 0.77 +/- 0.29 mL/g/10 min in the untreated ones. In contrast, neither NO formation after L-arginine supplementation nor inhibition of NO synthesis via the structural analogue (N(G)-monomethyl-L-arginine [L-NMMA]) changed cholestatic and glycogenolytic effects caused by norepinephrine. Only the sinusoidal bile acid uptake was reduced following increased NO formation. Thus, we conclude that endogenous NO formation prevents alpha-catecholaminergic-increased intrahepatic resistance without a major influence on the metabolic effects. PMID- 9214465 TI - Genetic and morphological findings in progressive familial intrahepatic cholestasis (Byler disease [PFIC-1] and Byler syndrome): evidence for heterogeneity. AB - Byler disease (ByD) is an autosomal recessive disorder in which cholestasis of onset in infancy leads to hepatic fibrosis and death. Children who have a clinically similar disorder, but are not members of the Amish kindred in which ByD was described, are said to have Byler syndrome (ByS). Controversy exists as to whether ByD and ByS (subtypes of progressive familial intrahepatic cholestasis [PFIC]) represent one clinicopathological entity. The gene for ByD has been mapped to a 19-cM region of 18q21-q22. PFIC caused by a lesion in this region, including ByD, can be designated PFIC-1. Examination of haplotypes in siblings with ByS in two unrelated non-Amish families showed that the gene(s) responsible for their disorder(s) did not lie in the PFIC-1 candidate region. On light microscopy and transmission electron microscopy (TEM), liver tissue differed between Amish children with PFIC-1, who had coarsely granular bile and at presentation had bland intracanalicular cholestasis, and the children with ByS in the two non-Amish families, who had amorphous or finely filamentous bile and at presentation had neonatal hepatitis. Bile acid composition of bile also differed: In the Amish children with PFIC-1 and in one ByS family, the proportional concentration of chenodeoxycholic acid (CDCA) in bile was low compared with normal bile; in the other ByS family, it was only slightly reduced. Genetic analysis and light microscopy and TEM of liver may help distinguish PFIC-1 from other forms of ByS. PMID- 9214466 TI - Extramedullary hematopoiesis in the adult mouse liver is associated with specific hepatic sinusoidal endothelial cells. AB - In previous work, two anatomically distinct-liver sinusoid endothelial cells (LEC): LEC-1 and LEC-2, have been described. We also reported that extramedullary hepatic hematopoiesis occurs only in close contact with LEC-1, suggesting that these cells may provide the microenvironment necessary for the maintenance and growth of hematopoietic cells. In the present work, we studied the capacity of LEC-1 and LEC-2 to maintain in vitro hematopoiesis. LEC-1 and LEC-2 were isolated and cloned from livers of adult mice. Bone marrow cells (BM) enriched with primitive hematopoietic progenitors were isolated from day-2, post-5-FU-treated mice (5-FUBMC). LEC-1 supported the maintenance and differentiation of hematopoietic progenitors for more than 6 weeks in vitro. In contrast, LEC-2 cells poorly supported the proliferation of hematopoietic cells for only two weeks of the co-culture. LEC-1 and 5-FUBMC cocultures showed cobblestone-area formation and the presence of hematopoietic progenitors that are able to form colonies (CFC) in the adhering fraction after six weeks of coculture. LEC-1 co cultures treated with a cocktail of cytokines (stem cell factor, interleukin [IL]1alpha, IL-3, and Epo) showed that megakaryocyte (CFU-Mk) and erythrocyte progenitors (BFU-e) were present during the entire period of the culture. Granulocyte-macrophage progenitors (CFU-GM) were present only during the first three weeks of the culture. These results suggest that LEC-1, but not LEC-2, provide an appropriate hematopoietic microenvironment for supporting the proliferation and differentiation of primitive hematopoietic cells. This could explain the anatomical restriction of hematopoietic cells for growing in LEC-1 domains during liver extramedullary hematopoiesis. PMID- 9214468 TI - Differential activation of heat shock and nuclear factor kappaB transcription factors in postischemic reperfused rat liver. AB - The aim of this study was to investigate the behavior of the transcription factors, heat-shock factor (HSF) and nuclear factor kappaB (NF-kappaB), in postischemic reperfused liver, with particular attention paid to possible differences in the time-course and mechanism of activation, which may help in defining their role in the response of the liver to reperfusion. Ischemia was induced by clamping the hilar pedicle of the left lateral and median liver lobes; the clamp was removed after 1 hour. Some rats were treated intraperitoneally with IL-1 receptor antagonist (IL-1RA) 30 minutes before ischemia and at the time of reperfusion. Binding of NF-kappaB to the corresponding consensus sequence is activated after 30 minutes of reperfusion, and is still increased 1 hour after reperfusion. Activation is suppressed in rats treated with IL-1RA; NF-kappaB persists in the cytosol associated with the inhibitor, IkappaB, and can be artifactually activated in vitro. Super-gel shift experiments revealed that the two subunits, p50 and p65, are involved in the activation of binding. In contrast, binding of HSF to the corresponding consensus sequence, heat shock element (HSE), is already activated at the end of ischemia, shows a further increase after 30 minutes of reperfusion, but declines 1 hour after reperfusion; more importantly, it is not inhibited by pretreatment of the rat with IL-1RA. In conclusion, although both HSF and NF-kappaB are activated by ischemia reperfusion, there are clear differences in time-course and mechanism of activation of the two transcription factors. Activation of HSF depends directly on some events occurring during ischemia; NF-kappaB is activated only after reperfusion and the concurrent oxidative stress, by an indirect mechanism that can be suppressed by IL-1RA. The possibility of dissociating the activation of these two transcription factors in postischemic reperfusion can have a prospective clinical relevance. PMID- 9214467 TI - Transferrin receptor recycling in rat hepatocytes is regulated by protein phosphatase 2A, possibly through effects on microtubule-dependent transport. AB - To understand the regulation of receptor-mediated endocytosis in hepatocytes, we have used two specific inhibitors of serine-threonine protein phosphatases (PP), microcystin (MCYST) and okadaic acid (OKA) as probes to alter protein phosphorylation in hepatocytes. We have then examined the impact of these changes on the specific binding and uptake of transferrin (Tf) in hepatocytes. The measurement of PP activity in hepatocyte lysates showed that OKA and MCYST shared a common inhibition of protein phosphatase 2A (PP2A). Our results showed that both OKA (250 nmol/L) and MCYST (500 nmol/L) significantly reduced Tf uptake at steady state (P < or = .05). The measurement of Tf internalization after 15 minutes in protein phosphatase inhibitor-pretreated cells revealed that the initial uptake was also significantly reduced. Binding studies showed that pretreatment with either of the phosphatase inhibitors did not result in significant changes in the K(d) for Tf binding to transferrin receptor (TfR). Additionally, no significant changes in the number of TfR in the plasma membrane were observed in phosphatase inhibitor-pretreated cells. The treatment of hepatocytes with nocodazole (NOC), which results in microtubule disassembly and inhibition of microtubule-based vesicle transport, caused comparable reductions in initial and steady state levels of transferrin accumulation. The changes in transferrin accumulation by both phosphatase inhibitors and nocodazole were accompanied by redistribution of the microtubule-anchored Golgi apparatus and lysosomal network from the perinuclear region to the cell periphery. Our data show that the regulation of Tf uptake by receptor-mediated endocytosis is mediated by PP2A and additionally may occur through regulation of microtubule based vesicle transport. PMID- 9214469 TI - Mini-microabscess syndrome in liver transplant recipients. AB - Cytomegalovirus (CMV) is a significant cause of morbidity in immunosuppressed patients. It is characterized in the liver by parenchymal microabscesses, usually containing CMV-infected cells. However, not all hepatic microabscesses are due to CMV infection. In 1992, we described "mini" microabscess (MMA) syndrome, a distinct clinical syndrome that occurs in transplanted livers. This report analyzes the clinical and laboratory features of 57 cases of MMA syndrome occurring in 52 patients and compares these with 19 biopsy-proven cases of CMV infection. The diagnosis of MMA syndrome can only be made histologically. The microabscesses are smaller and more numerous than in CMV infection, and there are no viral inclusions present. CMV DNA could not be detected in liver biopsy specimens with MMAs by using "nested" polymerase chain reaction (PCR), indicating that MMA syndrome is not caused by CMV infection. The pattern of liver enzyme and bilirubin elevation is predominantly hepatocellular, with transaminase levels elevated, on average, six to eight times the upper limit of normal. The clinical features of MMA syndrome are that it predominantly affects female (40 of 52 patients) orthotopic liver transplant (OLT) recipients of all ages (range, 11 months to 66.9 years). MMA syndrome is unrelated to the indication for initial OLT and tends to occur later after transplantation than CMV infection (median, 91 days post-OLT vs. 32 days for CMV hepatitis). Although the etiology of MMA syndrome is not clear, it does not appear to adversely affect graft or patient survival. PMID- 9214470 TI - Long-term consequence of rat orthotopic liver transplantation with and without hepatic arterial reconstruction: a clinical, pathological, and hemodynamic study. AB - Our aim was to investigate the time-related changes in various parameters following orthotopic rat liver transplantation with (AOLT) and without (NOLT) arterial reconstruction in male Lewis rats. Body weight and biochemical parameters were measured weekly, and a liver biopsy was obtained at 4, 8, and 12 weeks. Hemodynamics were evaluated at 12 weeks using the microsphere technique and compared with matched controls. Following AOLT, rats gained weight normally without any noticeable complication. In NOLT, two subgroups (NOLT-1 and NOLT-2) could clearly be identified retrospectively. In the NOLT-1 group, the body weight increased normally, although animals presented transient cholestasis. In these rats, the ductular proliferation found at 4 weeks had regressed by the 12th week with near-normal biopsies. By contrast, in the NOLT-2 group, rats did not gain body weight and had persistent cholestasis. Marked ductular proliferation with increasing fibrosis was observed, resulting in a secondary biliary cirrhosis by the 12th week. Surprisingly, rearterialization of the grafted liver occurred in both NOLT-1 and NOLT-2 irrespective of their clinical course. All transplanted rats showed portal hypertension with marked portosystemic shunts, probably caused by the portal cuff. However, a hyperdynamic circulatory state was only observed in the NOLT-2 group with cirrhotic changes. These findings further show the combined role of an intact hepatic innervation and of hepatocellular insufficiency in the genesis of the hyperdynamic circulatory state associated with portal hypertension. PMID- 9214471 TI - Thyroid abnormalities in chronic viral hepatitis and their relationship to interferon alfa therapy. AB - The prevalence of antithyroid peroxidase antibodies (ATPO) and/or of thyroid dysfunction was studied in 422 patients with chronic viral hepatitis C, B, and D. Baseline results were compared with those during and 6 months after interferon alfa (IFN-alpha) therapy. The overall prevalence of ATPO among untreated patients was 14.1%, with no significant differences between chronic hepatitis C, B, or D, as well as between males and females. However, high ATPO titers (> or = 18 IU/mL) clustered significantly among females (8.7% vs. 3.4%; P = .022), especially those with chronic hepatitis C (11.2% vs. 3.6%; P = .036). Before treatment, 3.7% of the patients had thyroid dysfunction, mostly hypothyroidism (3.5%), the latter increasing to 14.3% among patients with ATPO titers > or = 18 IU/mL. IFN-alpha treatment significantly increased overall thyroid dysfunction (9.7%; P = .001) and hypothyroidism (7.8%; P = .01), particularly among patients with high baseline ATPO (38.5%; P = .0002). Six months after stopping IFN-alpha treatment, the prevalence of thyroid dysfunction was 8.0%, still significantly higher than at baseline. By multivariate analysis, the only predictor positively associated with pre- or on-treatment hypothyroidism was the baseline titer of the ATPO antibodies (relative risk [RR], 3.0 and 3.8 per each log titer increase, respectively). In conclusion, patients with chronic viral hepatitis on IFN-alpha treatment exhibit an almost threefold increase of baseline thyroid dysfunction, persisting long after the end of therapy. High ATPO titers, clustering among females, particularly those with hepatitis C, represent the only predictor of pre and on-treatment hypothyroidism by multivariate analysis. Patients with chronic viral hepatitis, especially females, should be tested for ATPO and thyroid function and monitored during and posttreatment for free thyroxin (FT4) and thyroid-stimulating hormone (TSH) levels. PMID- 9214472 TI - Effect of lamivudine on morphology and function of mitochondria in patients with chronic hepatitis B. AB - Nucleoside analogues can induce mitochondrial dysfunction leading to severe clinical syndromes. Lamivudine, a new nucleoside analogue, is an active inhibitor of hepatitis B viral replication without apparent clinical toxicity. To assess subclinical mitochondrial toxicity, we studied the morphology and function of the mitochondrial system in 15 patients treated with lamivudine. Morphology was investigated by routine histological evaluation and electron-microscopic studies of mitochondria in liver biopsy specimens. Mitochondrial function was assessed by 2-keto[1-14C] isocaproic acid decarboxylation (KICA breath test) and by measuring the activity in liver biopsy specimens of the mitochondrial enzymes encoded by nuclear and mitochondrial DNA (mt-DNA) (complex I and IV) as well as a mitochondrial and a cytosolic enzyme both encoded by nuclear DNA only (complex II and lactic dehydrogenase [LDH]). All 15 patients underwent a liver biopsy before treatment and a KICA breath test before and during treatment; 13 agreed to undergo a repeat liver biopsy during lamivudine treatment. Liver tissue with no or minimal fibrotic changes from 7 patients treated for 6 months with lamivudine was suitable for assessment of the mitochondrial enzyme activity. We observed no signs of toxicity by routine histological or electron-microscopic evaluation. KICA breath tests revealed no differences in either peak exhalation or the area under the curve from 0 to 60 minutes between healthy controls (3.0% and 19.3%), untreated patients with chronic hepatitis B (3.4% and 19.3%), and patients treated with lamivudine (3.1% and 20.6%). The activities of the mt-DNA-encoded enzymes remained normal after lamivudine therapy. Unexpectedly a significant decrease in the activity of nuclear-DNA-encoded enzymes in patients with chronic hepatitis B in comparison with normal controls was found. The mean activity of complex II dropped from 45.3 to 20.0 micromol x min(-1), that of lactic dehydrogenase from 106 to 44 micromol x min(-1) (Wilcoxon rank sum; P < .05). In conclusion, no subclinical signs of mitochondrial toxicity resulting from lamivudine therapy for 6 months were observed. PMID- 9214473 TI - Synergistic inhibition of hepadnaviral replication by lamivudine in combination with penciclovir in vitro. AB - Lamivudine ([-]-beta-L-2',3'-dideoxy-3'-thiacytidine [3TC]) and penciclovir (9-[2 hydroxy-1-(hydroxymethyl)ethoxymethyl]guanine [PCV]) are potent inhibitors of hepatitis B virus (HBV) replication. Both drugs have entered phase III clinical trials for treatment of chronic HBV infection. 3TC and PCV are deoxycytidine and deoxyguanosine analogs, respectively, and their modes of action and how they interact are matters of both theoretical and practical interest. We compared the antiviral activities of 3TC and PCV alone and in combination in primary duck hepatocyte (PDH) cultures derived from ducklings congenitally infected with the duck hepatitis B virus (DHBV). 3TC and PCV inhibited DHBV replication to a comparable extent when used alone (50% inhibitory concentrations with 95% confidence intervals were 0.55 [0.50-0.59] micromol/L for 3TC and 0.35 [0.27 0.43] micromol/L for PCV), and in combination, the two nucleoside analogs acted synergistically over a wide range of clinically relevant concentrations. Synergy between PCV and 3TC was also observed in acutely infected cells and in "washout" experiments designed to assess the persistence of antiviral activity after drug removal. Furthermore, the combination was more effective in reducing the normally recalcitrant viral covalently closed circular (CCC) DNA form of DHBV than either drug alone. These results suggest that combinations of 3TC and PCV may act synergistically against HBV in vivo. PMID- 9214474 TI - Dose-dependent acute clearance of hepatitis C genotype 1 virus with interferon alfa. AB - To determine if the clearance of hepatitis C genotype 1 virus (HCV) is dependent on the dose of interferon alfa-2b (IFN-alpha2b), the acute clearance of HCV after a single dose of either 3, 5, or 10 mIU of IFN-alpha was compared in patients with chronic hepatitis C. HCV-RNA levels following IFN-alpha administration were measured. At 24 hours, mean percentage serum viral reduction was 41.4%, 63.7%, and 85.5% for 3, 5, and 10 mIU, respectively (P < .001). At 48 hours, the mean viral reduction was consistently less than the reduction at 24 hours, averaging 22.9%, 61.9%, and 74.3%, respectively (P < .001), indicating that the drug effect diminishes before 48 hours. Regression analysis showed a positive correlation between dose and percent reduction of HCV-RNA levels (r = .6; P < .001). A mathematical model showed that such dose dependence is expected if IFN-alpha partially blocks viral production. Minimum clearance and production rates of HCV were estimated from measurements of HCV-RNA levels after the 10-mIU dose. HCV decay followed an exponential decline with a minimum estimate of the viral clearance rate constant of 2.8 per day, corresponding to a virion half-life of 0.3 days or less. A minimal estimate of the daily HCV production and clearance is 3.7 x 10(11) virions per day, indicating a high rate of replication and turnover. These results indicate that there is a dose-dependent effect of IFN-alpha in clearance of HCV genotype 1. Because the virion production rate is very rapid and because the current recommended dose of IFN-alpha (3 mIU) is often ineffective, larger doses should be considered to treat genotype 1-infected patients. PMID- 9214475 TI - Does tumor necrosis factor play a role in alcoholic steatohepatitis? The potential pitfalls of a case-controlled allelic association analysis. PMID- 9214476 TI - Combination therapy for chronic hepatitis B. PMID- 9214477 TI - Liver injury associated with hepatitis C infection: is it the virus or is it the host? PMID- 9214478 TI - Early primary biliary cirrhosis: just delayed or different? PMID- 9214479 TI - A view of exocytotic vesicle fusion in living secretory cells. PMID- 9214481 TI - Acute liver failure in India. PMID- 9214480 TI - QT prolongation in end-stage liver disease: a result of altered sex hormone metabolism? PMID- 9214482 TI - High prevalence of hepatitis C virus infection in Japanese patients with porphyria cutanea tarda. PMID- 9214483 TI - Molecular biologists in Asia and Pacific plan research network. PMID- 9214484 TI - German law could boost prospects for organ transplants. PMID- 9214485 TI - US air pollution rules stir up a stink about research. PMID- 9214486 TI - Cloning for research 'should be allowed'. PMID- 9214487 TI - European biotech industry plans ethics panel. PMID- 9214488 TI - More danger in doctrine than in genetics. PMID- 9214489 TI - George Wald believed in apocalypse now. PMID- 9214490 TI - Alphabetical listing and citation rates. PMID- 9214491 TI - CJD transmission. PMID- 9214492 TI - Don't leave dignity out of the cloning debate. PMID- 9214493 TI - Predicting where we walk. PMID- 9214494 TI - Membrane fusion. Bridging the gap by AAA ATPases. PMID- 9214495 TI - Visual neurobiology. Colouring the cortex. PMID- 9214496 TI - Signal transduction. Mad about SMADs. PMID- 9214497 TI - Inhibition of ICE slows ALS in mice. PMID- 9214498 TI - Vision in dim light. PMID- 9214499 TI - A polymerase I palm in adenylyl cyclase? PMID- 9214500 TI - Detection of ozone on Saturn's satellites Rhea and Dione. AB - The satellites Rhea and Dione orbit within the magnetosphere of Saturn, where they are exposed to particle irradiation from trapped ions. A similar situation applies to the galilean moons Europa, Ganymede and Callisto, which reside within Jupiter's radiation belts. All of these satellites have surfaces rich in water ice. Laboratory studies of the interaction of charged-particle radiation with water ice predicted the tenuous oxygen atmospheres recently found on Europa and Ganymede. However, theoretical investigations did not anticipate the trapping of significantly larger quantities of O2 within the surface ice. The accumulation of detectable abundances of O3, produced by the action of ultraviolet or charged particle radiation on O2, was also not predicted before being observed on Ganymede. Here we report the identification of O3 in spectra of the saturnian satellites Rhea and Dione. The presence of trapped O3 is thus no longer unique to Ganymede, suggesting that special circumstances may not be required for its production. PMID- 9214501 TI - Modelling the evolution of human trail systems. AB - Many human social phenomena, such as cooperation, the growth of settlements, traffic dynamics and pedestrian movement, appear to be accessible to mathematical descriptions that invoke self-organization. Here we develop a model of pedestrian motion to explore the evolution of trails in urban green spaces such as parks. Our aim is to address such questions as what the topological structures of these trail systems are, and whether optimal path systems can be predicted for urban planning. We use an 'active walker' model that takes into account pedestrian motion and orientation and the concomitant feedbacks with the surrounding environment. Such models have previously been applied to the study of complex structure formation in physical, chemical and biological systems. We find that our model is able to reproduce many of the observed large-scale spatial features of trail systems. PMID- 9214502 TI - Endemic African mammals shake the phylogenetic tree. AB - The order Insectivora, including living taxa (lipotyphlans) and archaic fossil forms, is central to the question of higher-level relationships among placental mammals. Beginning with Huxley, it has been argued that insectivores retain many primitive features and are closer to the ancestral stock of mammals than are other living groups. Nevertheless, cladistic analysis suggests that living insectivores, at least, are united by derived anatomical features. Here we analyse DNA sequences from three mitochondrial genes and two nuclear genes to examine relationships of insectivores to other mammals. The representative insectivores are not monophyletic in any of our analyses. Rather, golden moles are included in a clade that contains hyraxes, manatees, elephants, elephant shrews and aardvarks. Members of this group are of presumed African origin. This implies that there was an extensive African radiation from a single common ancestor that gave rise to ecologically divergent adaptive types. 12S ribosomal RNA transversions suggest that the base of this radiation occurred during Africa's window of isolation in the Cretaceous period before land connections were developed with Europe in the early Cenozoic era. PMID- 9214503 TI - Colour tuning in human visual cortex measured with functional magnetic resonance imaging. AB - The primate retina contains three classes of cones, the L, M and S cones, which respond preferentially to long-, middle- and short-wavelength visible light, respectively. Colour appearance results from neural processing of these cone signals within the retina and the brain. Perceptual experiments have identified three types of neural pathways that represent colour: a red-green pathway that signals differences between L- and M-cone responses; a blue-yellow pathway that signals differences between S-cone responses and a sum of L- and M-cone responses; and a luminance pathway that signals a sum of L- and M-cone responses. It might be expected that there are neurons in the primary visual cortex with response properties that resemble these three perceptual pathways, but attempts to find them have led to inconsistent results. We have therefore used functional magnetic resonance imaging (fMRI) to examine responses in the human brain to a large number of colours. In visual cortical areas V1 and V2, the strongest response is to red-green stimuli, and much of this activity is from neurons receiving opposing inputs from L and M cones. A strong response is also seen with blue-yellow stimuli, and this response declines rapidly as the temporal frequency of the stimulus is increased. These responses resemble psychophysical measurements, suggesting that colour signals relevant for perception are encoded in a large population of neurons in areas V1 and V2. PMID- 9214504 TI - Hippocampal GABA(A) channel conductance increased by diazepam. AB - Benzodiazepines, which are widely used clinically for relief of anxiety and for sedation, are thought to enhance synaptic inhibition in the central nervous system by increasing the open probability of chloride channels activated by the inhibitory neurotransmitter gamma-aminobutyric acid (GABA). Here we show that the benzodiazepine diazepam can also increase the conductance of GABAA channels activated by low concentrations of GABA (0.5 or 5 microM) in rat cultured hippocampal neurons. Before exposure to diazepam, chloride channels activated by GABA had conductances of 8 to 53pS. Diazepam caused a concentration-dependent and reversible increase in the conductance of these channels towards a maximum conductance of 70-80 pS and the effect was as great as 7-fold in channels of lowest initial conductance. Increasing the conductance of GABAA channels tonically activated by low ambient concentrations of GABA in the extracellular environment may be an important way in which these drugs depress excitation in the central nervous system. That any drug has such a large effect on single channel conductance has not been reported previously and has implications for models of channel structure and conductance. PMID- 9214505 TI - p47 is a cofactor for p97-mediated membrane fusion. AB - At least two distinct ATPases, NSF and p97, are known to be involved in the heterotypic fusion of transport vesicles with their target membranes and the homotypic fusion of membrane compartments. The NSF-mediated fusion pathway is the best characterized, many of the components having been identified and their functions analysed. In contrast, none of the accessory proteins for the p97 mediated fusion pathway has been identified. Now we have identified the first such component, a protein of relative molecular mass 47,000 (p47), which forms a tight, stoichiometric complex with cytosolic p97 (one trimer of p47 per hexamer of p97). It is essential for the p97-mediated regrowth of Golgi cisternae from mitotic Golgi fragments, a process restricted to animal cells. As a homologue of p47 exists in budding yeast, this indicates that it might also be involved in other membrane fusion reactions catalysed by p97, such as karyogamy. PMID- 9214506 TI - Cofilin promotes rapid actin filament turnover in vivo. AB - The ability of actin filaments to function in cell morphogenesis and motility is coupled to their capacity for rapid assembly and disassembly. Because disassembly in vitro is much slower than in vivo, cellular factors that stimulate disassembly have long been assumed to exist. Although numerous proteins can affect actin dynamics in vitro, demonstration of in vivo relevance of these effects has not been achieved. We have used genetics and an actin-inhibitor in yeast to demonstrate that rapid cycles of actin assembly and disassembly depend on the small actin-binding protein cofilin, and that cofilin stimulates filament disassembly. These results may explain why cofilin is ubiquitous in eukaryotes and is essential for viability in every organism in which its function has been tested genetically. Magnitudes of disassembly defects in cofilin mutants in vivo were found to be correlated closely with the magnitudes of disassembly defects observed in vitro, supporting our conclusions. Furthermore, these cofilin mutants provided an opportunity to distinguish in living cells those actin functions that depend specifically on filament turnover (endocytosis) from those that do not (cortical actin patch motility). PMID- 9214507 TI - Mutations increasing autoinhibition inactivate tumour suppressors Smad2 and Smad4. AB - Smad2 and Smad4 are related tumour-suppressor proteins, which, when stimulated by the growth factor TGF-beta, form a complex to inhibit growth. The effector function of Smad2 and Smad4 is located in the conserved carboxy-terminal domain (C domain) of these proteins and is inhibited by the presence of their amino terminal domains (N domain). This inhibitory function of the N domain is shown here to involve an interaction with the C domain that prevents the association of Smad2 with Smad4. This inhibitory function is increased in tumour-derived forms of Smad2 and 4 that carry a missense mutation in a conserved N domain arginine residue. The mutant N domains have an increased affinity for their respective C domains, inhibit the Smad2-Smad4 interaction, and prevent TGF beta-induced Smad2 Smad4 association and signalling. Whereas mutations in the C domain disrupt the effector function of the Smad proteins, N-domain arginine mutations inhibit SMAD signalling through a gain of autoinhibitory function. Gain of autoinhibitory function is a new mechanism for inactivating tumour suppressors. PMID- 9214508 TI - A structural basis for mutational inactivation of the tumour suppressor Smad4. AB - The Smad4/DPC4 tumour suppressor is inactivated in nearly half of pancreatic carcinomas and to a lesser extent in a variety of other cancers. Smad4/DPC4, and the related tumour suppressor Smad2, belong to the SMAD family of proteins that mediate signalling by the TGF-beta/activin/BMP-2/4 cytokine superfamily from receptor Ser/Thr protein kinases at the cell surface to the nucleus. SMAD proteins, which are phosphorylated by the activated receptor, propagate the signal, in part, through homo- and hetero-oligomeric interactions. Smad4/DPC4 plays a central role as it is the shared hetero-oligomerization partner of the other SMADs. The conserved carboxy-terminal domains of SMADs are sufficient for inducing most of the ligand-specific effects, and are the primary targets of tumorigenic inactivation. We now describe the crystal structure of the C-terminal domain (CTD) of the Smad4/DPC4 tumour suppressor, determined at 2.5 A resolution. The structure reveals that the Smad4/DPC4 CTD forms a crystallographic trimer through a conserved protein-protein interface, to which the majority of the tumour-derived missense mutations map. These mutations disrupt homo oligomerization in vitro and in vivo, indicating that the trimeric assembly of the Smad4/DPC4 CTD is critical for signalling and is disrupted by tumorigenic mutations. PMID- 9214509 TI - DNA-replication checkpoint control at the Drosophila midblastula transition. AB - Embryogenesis is typically initiated by a series of rapid mitotic divisions that are under maternal genetic control. The switch to zygotic control of embryogenesis at the midblastula transition is accompanied by significant increases in cell-cycle length and gene transcription, and changes in embryo morphology. Here we show that mutations in the grapes (grp) checkpoint 1 kinase homologue in Drosophila block the morphological and biochemical changes that accompany the midblastula transition, lead to a continuation of the maternal cell cycle programme, and disrupt DNA-replication checkpoint control of cell-cycle progression. The timing of the midblastula transition is controlled by the ratio of nuclei to cytoplasm (the nucleocytoplasmic ratio), suggesting that this developmental transition is triggered by titration of a maternal factor by the increasing mass of nuclear material that accumulates during the rapid embryonic mitoses. Our observations support a model for cell-cycle control at the midblastula transition in which titration of a maternal component of the DNA replication machinery slows DNA synthesis and induces a checkpoint-dependent delay in cell-cycle progression. This delay may allow both completion of S phase and transcription of genes that initiate the switch to zygotic control of embryogenesis. PMID- 9214511 TI - A piece of my mind. Neighbors. PMID- 9214510 TI - Structure of the multimodular endonuclease FokI bound to DNA. AB - FokI is a member of an unusual class of bipartite restriction enzymes that recognize a specific DNA sequence and cleave DNA nonspecifically a short distance away from that sequence. Because of its unusual bipartite nature, FokI has been used to create artificial enzymes with new specificities. We have determined the crystal structure at 2.8A resolution of the complete FokI enzyme bound to DNA. As anticipated, the enzyme contains amino- and carboxy-terminal domains corresponding to the DNA-recognition and cleavage functions, respectively. The recognition domain is made of three smaller subdomains (D1, D2 and D3) which are evolutionarily related to the helix-turn-helix-containing DNA-binding domain of the catabolite gene activator protein CAP. The CAP core has been extensively embellished in the first two subdomains, whereas in the third subdomain it has been co-opted for protein-protein interactions. Surprisingly, the cleavage domain contains only a single catalytic centre, raising the question of how monomeric FokI manages to cleave both DNA strands. Unexpectedly, the cleavage domain is sequestered in a 'piggyback' fashion by the recognition domain. The structure suggests a new mechanism for nuclease activation and provides a framework for the design of chimaeric enzymes with altered specificities. PMID- 9214512 TI - The effect of universal health insurance on health care utilization in Taiwan. Results from a natural experiment. AB - CONTEXT: The government of Taiwan introduced universal health insurance to cover all citizens in 1995. This national health insurance program was proposed to assure the accessibility to health care at reasonable cost. Evaluation of the consequences, including health care utilization and expenditure, is crucial for policy adjustment. OBJECTIVES: To evaluate the effect of Taiwan's national health insurance on health care utilization. DESIGN: Cohort survey conducted before and after the implementation of the national health insurance program. PARTICIPANTS: A total of 1021 randomly selected Taiwanese adults. MAIN OUTCOME MEASURES: Physician visits in the 2 weeks prior to the survey and hospital admissions and emergency department visits in the immediate past year. RESULTS: After the introduction of universal health insurance, the newly insured consumed more than twice the amount of outpatient physician visits (0.21 vs 0.48, P<.05) and hospital admissions (0.04 vs 0.11, P<.05) than before universal health insurance was implemented, bringing them to the same amount of health care contacts as the previously insured group. The newly insured also experienced an insignificant increase in emergency department visits. In contrast, the previously insured group had a small but statistically significant increase in outpatient visits (0.48 vs 0.59, P<.05) and insignificant changes in hospital admissions and emergency department visits. CONCLUSION: The universal health insurance removed some barriers to health care for those newly insured. The copayment design in the insurance scheme seemed to have an insignificant effect on curbing medical care utilization. Taiwanese health policy analysts should seriously consider the growth of health care expenditures since the implementation of universal health insurance. PMID- 9214513 TI - Campaign launched to tell physicians, public about emergency contraception. PMID- 9214514 TI - From Rx to OTC: more drugs make the switch. PMID- 9214515 TI - Estrogen trial for patients with endometrial cancer. PMID- 9214516 TI - From the Centers for Disease Control and Prevention. Update: outbreaks of cyclosporiasis--1997. PMID- 9214517 TI - From the Centers for Disease Control and Prevention. Lactic acidosis traced to thiamine deficiency related to nationwide shortage of multivitamins for total parenteral nutrition--United States, 1997. PMID- 9214518 TI - From the Centers for Disease Control and Prevention. Lyme disease--United States, 1996. PMID- 9214519 TI - Brain infarction and the clinical expression of Alzheimer disease. PMID- 9214520 TI - Brain infarction and the clinical expression of Alzheimer disease. PMID- 9214521 TI - Angioplasty vs bypass surgery in patients with multivessel coronary artery disease. PMID- 9214522 TI - Preventing hypothermia-related death. PMID- 9214523 TI - Clinical crossroads update: an 82-year-old woman with cataracts. PMID- 9214524 TI - Magazines containing tobacco advertisements: proscription not subscription. PMID- 9214525 TI - Sexually transmitted diseases in disadvantaged Australian communities. PMID- 9214526 TI - Outcomes of stroke patients in Medicare fee for service and managed care. AB - CONTEXT: Increasing numbers of Medicare beneficiaries have been enrolling in health maintenance organizations (HMOs) because HMO participation reduces out-of pocket expenses, and the federal government views HMOs as a way to contain Medicare costs. However, results comparing outcomes and quality of care in HMOs vs fee for service (FFS) have been mixed, and outcomes after stroke have not been adequately assessed. OBJECTIVE: To compare discharge destinations and survival rates following stroke in Medicare HMOs with similar FFS settings. DESIGN: An observational study for 2 groups evaluating stroke patients' discharge destinations and survival times from the date of hospital admission. SETTING: A total of 19 HMOs were selected from 12 states. The FFS sample was drawn from the same geographic areas. PATIENTS: The sample included 402 HMO patients from 71 hospitals and 408 FFS patients from 60 hospitals. PROCESS AND OUTCOME MEASURES: Data were abstracted from medical records on demographics, clinical characteristics of stroke, comorbid illnesses, and discharge destinations following hospitalization. Data on survival were obtained from Medicare files and included 25 to 37 months of follow-up (median, 30.4 months, HMO; 31.1 months, FFS) from the date of hospital admission. RESULTS: There were 109 patients who died during the hospitalization (49 HMO, 12.2%; 60 FFS, 14.7%), and a total of 410 patients had died by the end of follow-up (191 HMO, 47.5%; 219 FFS, 53.7%). Approximately one fourth of both groups had do-not-resuscitate orders (HMO, 25.4%; FFS, 27.9%; P=.68). After controlling for age, marital status, and characteristics of dependency at discharge, HMO patients were more likely than FFS patients to be sent to nursing homes (HMO, 41.8%; FFS, 27.9%; P=.001) and less likely to be discharged to rehabilitation hospitals or units (HMO, 16.2%; FFS, 23.4%; P=.03). At follow-up, no significant differences in relative risk of dying were found between HMO and FFS groups (relative risk, 0.96; 95% confidence interval, 0.73-1.26; P=.77). CONCLUSIONS: Patients in Medicare HMOs who experience strokes are more likely to be discharged to nursing homes and less likely to go to rehabilitation facilities following the acute event. However, they have similar survival patterns compared with comparable patients in FFS settings after adjusting for other factors. PMID- 9214527 TI - Safety and efficacy of pramipexole in early Parkinson disease. A randomized dose ranging study. Parkinson Study Group. AB - CONTEXT: Monotherapy with dopamine agonists may be useful in early Parkinson disease. OBJECTIVE: To evaluate dose-response relationships for tolerability, safety, and efficacy of the synthetic dopamine agonist pramipexole. DESIGN: Multicenter, multidosage, parallel-group, double-blind, placebo-controlled, randomized clinical trial. SETTING: University or academically based movement disorder clinics. PATIENTS: A total of 264 patients with early Parkinson disease (PD) who were not requiring or receiving levodopa or other dopamine agonists were enrolled. INTERVENTION: Subjects were randomized to 1 of 5 treatment groups: pramipexole doses of 1.5 mg/d, 3.0 mg/d, 4.5 mg/d, and 6.0 mg/d, or matching placebo. A 6-week dosage escalation period was followed by a 4-week maintenance period and a 1-week period during which active treatment was withdrawn. MAIN OUTCOME MEASURES: The primary measure of tolerability was the proportion of subjects completing the study on the assigned treatment. The primary measure of efficacy was the change from baseline to 10 weeks in the total score on the Unified Parkinson's Disease Rating Scale (UPDRS). RESULTS: Pramipexole was generally safe and well tolerated in this 10-week study. The proportion of subjects completing the study on the originally assigned dosage was 98% for placebo and 81% for the 1.5-mg/d, 92% for the 3.0-mg/d, 78% for the 4.5-mg/d, and 67% for the 6.0-mg/d treatment groups. There was a trend toward increased frequency of adverse experiences, particularly somnolence, in the 6.0-mg/d group. After 10 weeks of treatment, pramipexole-treated subjects showed a 20% improvement in total UPDRS scores, with mean improvements in scores ranging from 5.9 to 7.0 units among active treatment groups, compared with 0.9 units for the placebo group (P<.005 for each comparison with placebo). There was also evidence that the treatment effects were more pronounced in subjects with worse UPDRS scores at baseline. CONCLUSIONS: Pramipexole is safe and effective as short-term monotherapy in patients with early PD who are not receiving levodopa. Further study is warranted to determine the long-term impact of pramipexole on the progression of disability in PD and its value in comparison with levodopa therapy and other dopamine agonists. PMID- 9214528 TI - Prevalence of child physical and sexual abuse in the community. Results from the Ontario Health Supplement. AB - CONTEXT: Although child maltreatment is considered common, few community surveys have examined the prevalence of more than 1 type of maltreatment among both males and females. OBJECTIVE: To determine the prevalence of a history of physical and sexual abuse during childhood among the general population. DESIGN: General population survey. SETTING: Household dwellings in the province of Ontario, Canada. PARTICIPANTS: A random sample (N=9953) of residents aged 15 years and older participated in the Ontario Health Supplement. MAIN OUTCOME MEASURE: Self administered questionnaire about a history of physical and sexual abuse in childhood. RESULTS: A history of child physical abuse was reported more often by males (31.2%) than females (21.1%), while sexual abuse during childhood was more commonly reported by females (12.8%) than males (4.3%). Severe physical abuse was reported by similar proportions of males (10.7%) and females (9.2%). A greater percentage of females reported a history of severe sexual abuse (11.1%) compared with males (3.9%). Age of the respondent was not significantly associated with childhood abuse within any category for males. However, for females, the reported prevalence in childhood of sexual abuse, co-occurrence of physical and sexual abuse, and both categories of severe abuse decreased with increasing age of the respondent. CONCLUSIONS: A history of childhood maltreatment among Ontario residents is common. Child abuse may be more prevalent in younger women compared with older women, or there may be a greater willingness among younger women to report abuse. PMID- 9214529 TI - Apolipoprotein E epsilon4 associated with chronic traumatic brain injury in boxing. AB - CONTEXT: Given the similarities between Alzheimer disease and dementia pugilistica, we evaluated the relationship between apolipoprotein E (APOE) genotype and chronic traumatic brain injury (CTBI) in boxers to determine whether there is a genetic susceptibility to the effects of head trauma. OBJECTIVE: To assess the relationship between CTBI and APOE genotype in boxers. DESIGN AND SETTING: Clinical characterization of 24 volunteer and 6 referred boxers in an outpatient setting. PARTICIPANTS: Thirty professional boxers aged 23 to 76 years underwent neurologic and behavioral assessment in conjunction with APOE genotyping. MAIN OUTCOME MEASURES: Apolipoprotein E genotype was examined in relationship to measures of CTBI. A 10-point clinical rating scale (0-9), the Chronic Brain Injury (CBI) scale, was devised to assess the severity of traumatic encephalopathy associated with boxing. Boxers with abnormal CTBI scores were further classified on the basis of whether their impairments were possibly or probably related to boxing. Scores were analyzed in relation to boxing exposure (number of bouts) and APOE genotype. RESULTS: Among the 30 boxers, 11 were found to be normal (CBI score=0), 12 showed mild deficits (CBI score=1-2), 4 were moderately impaired (CBI score=3-4), and 3 showed signs of severe impairment (CBI score > 4). High-exposure boxers (ie, those with > or = 12 professional bouts) had significantly higher CBI scores (mean [SD], 2.6 [1.9]) than low-exposure boxers (mean [SD], 0.3 [0.7]) (P<.001), indicating that neurologic impairment as measured by the CBI scale seems related to boxing exposure. The APOE genotype frequencies of the study population were approximately the same as those found in the general population. Boxers with low exposure had mean CBI scores of 0.33, irrespective of APOE genotype. However, high-exposure boxers with an APOE epsilon4 allele had significantly greater CBI scores (mean [SD], 3.9 [2.3]) than high-exposure boxers without APOE epsilon4 (mean [SD], 1.8 [1.2]) (P=.04). All boxers with severe impairment possessed at least 1 APOE epsilon4 allele. The tendency for greater CTBI among those with both high exposure and an epsilon4 allele was statistically significant at the P<.001 level. CONCLUSIONS: These preliminary findings suggest that possession of an APOE epsilon4 allele may be associated with increased severity of chronic neurologic deficits in high exposure boxers. PMID- 9214530 TI - Personal use of drug samples by physicians and office staff. AB - CONTEXT: Pharmaceutical samples are commonly used in ambulatory care settings. There is limited research on their use or impact on health care providers and patients. OBJECTIVE: To determine the extent of personal use of drug samples over a 1-year period by physicians and medical office staff. DESIGN, SUBJECTS, AND SETTING: An anonymous cross-sectional survey of all physicians, resident physicians, nursing staff, and office staff in a family practice residency. MAIN OUTCOME MEASURE: Quantity of drug samples taken for personal or family use. RESULTS: Of 55 surveys issued, 53 (96%) were returned. A total of 230 separate drug samples were reported taken in amounts ranging from 1 dose to greater than 1 month's supply. Two respondents reported no use of drug samples, while 4 respondents reported taking more than 10 different samples. CONCLUSION: Drug samples are commonly taken by physicians and office staff for personal and family use. The ethical implications of this practice warrant further discussion. PMID- 9214531 TI - Pharmacological management of alcohol withdrawal. A meta-analysis and evidence based practice guideline. American Society of Addiction Medicine Working Group on Pharmacological Management of Alcohol Withdrawal. AB - OBJECTIVE: To provide an evidence-based practice guideline on the pharmacological management of alcohol withdrawal. DATA SOURCES: English-language articles published before July 1, 1995, identified through MEDLINE search on "substance withdrawal--ethyl alcohol" and review of references from identified articles. STUDY SELECTION: Articles with original data on human subjects. DATA ABSTRACTION: Structured review to determine study design, sample size, interventions used, and outcomes of withdrawal severity, delirium, seizures, completion of withdrawal, entry into rehabilitation, adverse effects, and costs. Data from prospective controlled trials with methodologically sound end points corresponding to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, were abstracted by 2 independent reviewers and underwent meta-analysis. DATA SYNTHESIS: Benzodiazepines reduce withdrawal severity, reduce incidence of delirium (-4.9 cases per 100 patients; 95% confidence interval, -9.0 to -0.7; P=.04), and reduce seizures (-7.7 seizures per 100 patients; 95% confidence interval, -12.0 to -3.5; P=.003). Individualizing therapy with withdrawal scales results in administration of significantly less medication and shorter treatment (P<.001). beta-Blockers, clonidine, and carbamazepine ameliorate withdrawal severity, but evidence is inadequate to determine their effect on delirium and seizures. Phenothiazines ameliorate withdrawal but are less effective than benzodiazepines in reducing delirium (P=.002) or seizures (P<.001). CONCLUSIONS: Benzodiazepines are suitable agents for alcohol withdrawal, with choice among different agents guided by duration of action, rapidity of onset, and cost. Dosage should be individualized, based on withdrawal severity measured by withdrawal scales, comorbid illness, and history of withdrawal seizures. beta Blockers, clonidine, carbamazepine, and neuroleptics may be used as adjunctive therapy but are not recommended as monotherapy. PMID- 9214532 TI - Electronic communication with patients. Evaluation of distance medicine technology. AB - OBJECTIVE: To evaluate controlled evidence on the efficacy of distance medicine technologies in clinical practice and health care outcome. DATA SOURCES: Systematic electronic database and manual searches (1966-1996) were conducted to identify clinical trial reports on distance medicine applications. STUDY SELECTION: Three eligibility criteria were applied: prospective, contemporaneously controlled clinical trial with random assignment of the intervention; electronic distance technology application in the intervention group and no similar intervention in the control group; and measurement of the intervention effect on process or outcome of care. DATA EXTRACTION: Data were abstracted by independent reviewers using a standardized abstraction form and the quality of methodology was scored. Distance technology applications were described in 6 categories: computerized communication, telephone follow-up and counseling, telephone reminders, interactive telephone systems, after-hours telephone access, and telephone screening. DATA SYNTHESIS: Of 80 eligible clinical trials, 61 (76%) analyzed provider-initiated communication with patients and 50 (63%) reported positive outcome, improved performance, or significant benefits, including studies of computerized communication (7 of 7), telephone follow-up and counseling (20 of 37), telephone reminders (14 of 23), interactive telephone systems (5 of 6), telephone access (3 of 4), and telephone screening (1 of 3). Significantly improved outcomes were demonstrated in studies of preventive care, management of osteoarthritis, cardiac rehabilitation, and diabetes care. CONCLUSIONS: Distance medicine technology enables greater continuity of care by improving access and supporting the coordination of activities by a clinician. The benefits of distance technologies in facilitating communication between clinicians and patients indicate that application of telemedicine should not be limited to physician-to-physician communication. PMID- 9214533 TI - Humanitarianism under the gun. PMID- 9214534 TI - Stroke patients, "managed care," and distributive justice. PMID- 9214535 TI - Crisis, ethics, and the American Medical Association 1847 and 1997. PMID- 9214536 TI - Cystic parotid gland enlargement in HIV disease. The diffuse infiltrative lymphocytosis syndrome. PMID- 9214537 TI - Neurochemical architecture of the human striatum. AB - The striatum of the human brain has a highly differentiated neurochemical architecture visible in stains for many of the neurotransmitter-related molecules present in the striatum. The distributions for these chemical markers have never been analyzed comprehensively. We compared the distributions of multiple neurochemical markers in a serial-section analysis of the caudate nucleus, the putamen, and the ventral striatum in normal human brains. The cholinergic system was identified with choline acetyltransferase (ChAT). The organization of the cholinergic fiber system was compared with that of striatal systems expressing immunoreactivity for calbindin D28k, met-enkephalin, substance P, tyrosine hydroxylase, and parvalbumin. Each striatal region analyzed displayed a unique neurochemical organization. In the dorsal caudate nucleus, the distribution of all markers followed the classical striosome/matrix organization as previously reported. In the dorsal putamen, ChAT-staining was less intense, and striosomes were delineated primarily by unstained fiber bundles. In the ventral caudate nucleus/nucleus accumbens region, the boundaries of ChAT-stained regions were not always visible with stains for calbindin, enkephalin, and substance P. The ventral putamen displayed a similar organization, except in its lateral part, where ChAT-poor regions were often found adjacent to, rather than in register with, regions expressing low levels of the other markers (calbindin, enkephalin, substance P, and tyrosine hydroxylase). Our findings suggest that, in addition to the classical striosome-matrix organization visible in the dorsal caudate nucleus and putamen, there is further neurochemical differentiation in a large ventral part of the caudate nucleus and putamen and in the ventral striatum-nucleus accumbens proper. The more complex relationships among the different neurochemical systems in the ventral striatum may reflect the increase in size in the primate of striatal regions associated with association and limbic cortex. PMID- 9214538 TI - Development of primary visual projections occurs entirely postnatally in the fat tailed dunnart, a marsupial mouse, Sminthopsis crassicaudata. AB - We have examined the development of retinal projections in a diminutive polyprotodont marsupial, the fat-tailed dunnart, Sminthopsis crassicaudata. Here, we document the most immature mammalian visual system at birth described to date. At postnatal day (P) 0, the retinal ganglion cell layer has yet to form, and axons have not entered the optic stalk. By P4, the retinal ganglion cell layer could be distinguished at the posterior pole, and the front of growing axons extended one-third the length of the optic stalk, a distance of approximately 150 microm; a few pioneer growth cones had grown beyond the main axon group but had still to reach the midline. Axons had decussated at the optic chiasm by P10 to penetrate the base of the contralateral optic tract and, by P15, had reached the dorsal lateral geniculate nucleus (dLGN), superior colliculus (SC), and accessory optic system (AOS); ipsilaterally projecting axons matured slightly later. From P20, axons had reached the caudal SC both contralaterally and ipsilaterally and terminated throughout the depth of the retinorecipient layers. After P30, the projections gradually refined. Within the rostral dLGN, segregation into four contralateral and four ipsilateral bands occurred by P50, approximately 5 days after eye opening. The projection to the ipsilateral SC underwent refinement by P50, becoming restricted to its rostral pole, and presented as discrete patches within the stratum opticum. At birth, the dunnart visual system is comparable to early to midembryonic stages [embryonic day (E) 12, E14, E19, E24, and E30, respectively] in the mouse, rat, ferret, cat, and monkey. The extreme immaturity of the neonatal dunnart together with the observation that the entire development of the primary optic pathway occurs postnatally over a protracted period make this marsupial especially valuable for investigating factors that control pathway formation in the early developing mammalian primary visual system. PMID- 9214539 TI - Substance P innervation of the rat hippocampal formation. AB - Light and electron microscopic substance P (SP) immunostaining was performed on hippocampal sections of colchicine-pretreated, control, untreated fimbria-fornix transected (5 days), as well as perforant path-stimulated Sprague-Dawley rats fixed in 5% acrolein. Numerous SP-immunoreactive neurons could be observed in the stratum oriens of the Ammon's horn and subiculum, fewer were seen in the dentate hilar area and stratum radiatum of CA2 and CA3, and even fewer were seen at the border between the CA1 strata radiatum and the lacunosum moleculare of CA1 subfield. A higher dose of colchicine resulted in SP immunoreactivity in a large population of granule cells and mossy axon terminals. The entire CA2 region, the stratum oriens of CA1, CA3, and the subiculum were densely innervated by SP containing axon terminals. A homogeneous SP innervation was found in the stratum radiatum of CA1. Only a few SP fibers were seen adjacent to the granule cell layer. Symmetric axosomatic contacts were seen between SP-containing boutons and somata in the stratum oriens of the Ammon's horn. However, throughout the hippocampal formation, the majority of SP-containing axons formed axodendritic symmetric synapses. A dense population of SP-immunoreactive boutons that formed axodendritic asymmetric synapses was observed in the strata oriens and radiatum of the CA3a and CA2 regions, and a few were found in the supragranular and subgranular layers of the dentate gyrus. Fimbria-fornix transection resulted in a marked loss of SP fibers in the strata oriens, pyramidale, and radiatum of the CA3a and CA2 subfields. In perforant pathway-stimulated animals, a population of granule cells and a large number of mossy axon terminals were immunoreactive for SP. These observations suggest two sources of SP innervation to the hippocampal formation: one arising from intrinsic sources (interneurons and granule cells) and one arising from extrinsic sources, most likely the supramammillary region. PMID- 9214540 TI - Collateral projections of single neurons in the posterior thalamic region to both the temporal cortex and the amygdala: a fluorescent retrograde double-labeling study in the rat. AB - It has been reported that the acoustic thalamus of the rat sends projection fibers to both the temporal cortical areas and the lateral amygdaloid nucleus to mediate conditioned emotional responses to an acoustic stimulus. In the present study, fluorescent retrograde double labeling with Fast Blue and Diamidino Yellow has been used in the rat to examine whether single neurons in the posterior thalamic region send axon collaterals to both the temporal cortical areas and lateral amygdaloid nucleus. One of the tracers was injected into the lateral amygdaloid nucleus and the other into the temporal cortical areas close to the rhinal sulcus. Neurons double-labeled with both tracers were found mainly in the posterior intralaminar nucleus and suprageniculate nucleus, and to a lesser extent in the subparafascicular nucleus and medial division of the medial geniculate nucleus. No double-labeled neurons were seen in either the dorsal or ventral division of the medial geniculate nucleus. When one of the tracers was injected into the lateral amygdaloid nucleus and the other into either the dorsal portion of the temporal cortex, the dorsal portion of the entorhinal cortex, or the posterior agranular insular cortex, no double-labeled neurons were found in the posterior thalamic region. The present results indicate that a substantial number of single neurons in the acoustic thalamus project to both the limbic cortical areas and lateral amygdaloid nucleus by way of axon collaterals. These neurons may be implicated in affective and autonomic components of responses to multi-sensory stimuli, including acoustic ones. PMID- 9214541 TI - Differential central projections of physiologically characterized horizontal semicircular canal vestibular nerve afferents in the toadfish, Opsanus tau. AB - Anatomical and neurophysiological studies were undertaken to examine the central projection pattern of physiologically characterized horizontal semicircular canal vestibular nerve afferents in the toadfish, Opsanus tau. The variations in individual response characteristics of vestibular nerve afferents to rotational stimulus provided a means of typing the afferents into descriptive classes; the afferents fell into a broad continuum across the spectrum from low-gain, velocity sensitive to high-gain, acceleration-sensitive responses (Boyle and Highstein [1990b] J. Neurosci. 10:1557-1569; Boyle and Highstein [1990a] J. Neurosci. 10:1570-1582). In the present study, each afferent was typed as a low-gain, high gain, or acceleration fiber during rotational or mechanical stimulation (Rabbitt et al. [1995] J. Neurophysiol. 73:2237-2260) and was then intracellularly injected with biocytin. The axons were reconstructed, and the morphology, synaptic boutons, and projection pattern of each axon were determined. The results indicated that the three descriptive classes of vestibular nerve afferents have unique as well as overlapping central projection patterns and destinations in the vestibular nuclei, with intranuclear parcellation in the anterior octavus, magnocellularis, tangentialis, posterior octavus, and descending octavus nuclei. In general, increased sensitivity and faster response dynamics were correlated with both a more extensive central projection and a progressive increase in morphological complexity. Low-gain, velocity-sensitive fibers were the simplest morphologically, with the fewest number of branches (n = 17) and shortest length (4,282 microm), and projections were confined to the middle portions of the vestibular nuclei. High-gain, velocity-sensitive fibers were morphologically more diverse than low-gain fibers, with a greater number of branches (n = 26), longer length (6,059 microm), 29% greater volume, and a more widespread projection pattern with projections to both the anterior and the middle portions of the vestibular nuclei. Acceleration fibers were morphologically distinct from low- and high-gain fibers, with more elaborate branching (n = 41), greatest overall length (17,370 microm) and volume (16% greater than high gains), and displayed the most extensive central projection pattern, innervating all vestibular nuclei except tangentialis. Thus, there are anatomically demonstrable differential central projections of canal afferents with different response dynamics within the vestibular complex of the fish. PMID- 9214543 TI - Local circuit neurons of macaque monkey striate cortex: IV. Neurons of laminae 1 3A. AB - We continue our Golgi studies (Lund [1987] J. Comp. Neurol. 257:60-92; Lund et al. [1988] J. Comp. Neurol. 276:1-29; Lund and Yoshioka [1991] J. Comp. Neurol. 331:234-258) of the organization of local circuit, largely gamma-aminobutyric acid (GABA)-containing neurons in macaque monkey visual cortex, area V1, with this account of the local circuit neurons lying in layers 1 and 2/3A. These layers receive intrinsic interlaminar excitatory and inhibitory relays from layers 3B, 4A, 4B, and 5. We describe seven varieties of local circuit neurons with somata within layers 1-2/3A, and we compare the lateral scale of spread of the axons and dendrites of these neurons with the size of the columnar connectional patch domains made by the laterally spreading axon collaterals of pyramidal neurons within the superficial layers (Lund et al. [1993] Cerebral Cortex 3:148-162). We conclude from this comparison that all of the neurons have dendritic fields that are limited to single patch domains. Furthermore, only two of the seven local circuit neuron varieties have sufficient axon spread to influence territory beyond single domains, reaching into neighboring territory likely to differ in function from that occupied by their dendrites. We have identified descending projections from particular varieties to layers 3B, 4A, 4B, and 5 and to the white matter. We discuss the contributions that these interneurons may make to function within the superficial cortical layers, and we summarize our overall conclusions, so far, from our set of studies on interneurons within area V1 of the macaque. PMID- 9214542 TI - Evidence for a possible avian dorsal thalamic region comparable to the mammalian ventral anterior, ventral lateral, and oral ventroposterolateral nuclei. AB - In the present study, we investigated whether a dorsal thalamic region comparable to the motor part of the mammalian ventral tier (the ventral anterior nucleus, the ventral lateral nucleus, and the oral ventroposterolateral nucleus) exists in pigeon. With this aim, we reinvestigated the projections of the pigeon dorsal pallidum to the dorsal thalamus by using 1) injections of the anterogradely transported form of biotinylated dextran amine (BDA; 10,000 molecular weight) in the pigeon dorsal pallidum (paleostriatum primitivum) and 2) injections of the retrogradely transported form of BDA (3,000 molecular weight) in the pigeon dorsal thalamus. Our results indicate that the dorsal pallidum in pigeons projects to three areas of the dorsal thalamus: the dorsointermediate posterior nucleus, the ventrointermediate area, and the nucleus subrotundus. Only the projection to the dorsointermediate posterior nucleus was described previously (Karten and Dubbeldam [1973] J. Comp. Neurol. 148:61-90; Kitt and Brauth [1982] Neuroscience 6:1551-1566). To investigate whether any of the dorsal thalamic nuclei receiving pallidal input project to a motor cortical field, injections of the retrograde tracer Fluoro-Gold were placed into the rostral Wulst. This is an avian cortical field that appears to contain a region comparable to mammalian primary somatomotor cortex (Karten [1971] Anat. Rec. 169:353; Wild [1992] J. Comp. Neurol. 287:1-18). Our results indicate that neurons in the rostral ventrointermediate area, but not in the nucleus subrotundus, the dorsointermediate posterior nucleus, or the intermediate or caudal parts of the ventrointermediate area, project to the rostral Wulst. In addition to the input from the dorsal pallidum, the avian ventrointermediate area also receives input from the lateral substantia nigra and the lateral and internal cerebellar nuclei (present results). Our results suggest the existence in birds of a pallidothalamocortical loop similar to the pallidoventral tier-motor cortex loop of mammals and suggest that the avian ventrointermediate area is comparable to the motor part of the mammalian ventral tier in both location and connections. If this is confirmed by physiological experiments, then it would indicate that basal ganglia control of movement mediated by a pallidothalamocortical loop may have evolved with the stem reptiles. PMID- 9214544 TI - Association of serotonin-like immunoreactive axons with nociceptive projection neurons in the caudal spinal trigeminal nucleus of the rat. AB - Serotoninergic projections to the spinal dorsal horn are implicated in the modulation of nociceptive transmission. However, morphological evidence indicating that serotoninergic projection fibers make synapses on nociceptive neurons in the medullary dorsal horn is still meager. Thus, we examined whether axonal varicosities with serotonin (5-HT)-like immunoreactivity (5-HT-LI) might make synapses on nociceptive projection neurons in the caudal spinal trigeminal nucleus (Vc) of the rat. Projection neurons were retrogradely labeled with tetramethylrhodamine-dextran amine (TMR-DA) or wheat germ agglutinin-horseradish peroxidase (WGA-HRP) that was injected into the parabrachial or thalamic region. Vc neurons in which c-fos protein-like immunoreactivity (Fos-LI) was induced by subcutaneous injection of formalin into the lip were considered nociceptive. Vc neurons in direct contact with axonal varicosities that bind isolectin I-B4 were also considered nociceptive. Triple labeling for 5-HT, TMR-DA, and Fos as well as that for 5-HT, TMR-DA, and I-B4 were done by using the immunofluorescence and fluorescence histochemical techniques. Confocal laser-scanning microscopy revealed that axonal varicosities with 5-HT-LI were in close apposition to TMR-DA labeled neurons showing Fos-LI in lamina I and the outer part of lamina II (lamina IIo), and that both axonal varicosities with 5-HT-LI and those binding I B4 were in close apposition to single neuronal profiles labeled with TMR-DA. The presumed nociceptive neuronal profiles in close apposition to axon terminals with 5-HT-LI were mainly those of laminae I and II neurons as well as dendrites of lamina III neurons. Electron microscopy confirmed that axon terminals with 5-HT LI and those with I-B4 binding activity in laminae I and II made synapses on somatic and dendritic profiles that were labeled with WGA-HRP. The results indicate that serotoninergic neurons project directly on nociceptive projection neurons in the Vc. PMID- 9214545 TI - Projections of the sexually dimorphic anteroventral periventricular nucleus in the female rat. AB - The anteroventral periventricular nucleus of the hypothalamus (AVPV) is a sexually dimorphic nucleus in the preoptic region that appears to be a nodal point in forebrain circuits, mediating hormonal feedback on gonadotropin secretion. The results of anterograde transport experiments indicate that the AVPV sends ascending projections to the ventral part of the lateral septal nucleus, the parastrial nucleus, and the region adjacent to the vascular organ of the lamina terminalis (OVLT) that contains a subpopulation of gonadotropin releasing hormone (GnRH)-containing neurons. The majority of projections from the AVPV pass caudally through the periventricular zone of the hypothalamus and form dense terminal fields in the periventricular nuclei, parvicellular parts of the paraventricular nucleus, and in the arcuate nucleus. Inputs to medial zone nuclei are more limited, with substantial projections to only the medial preoptic and dorsomedial nuclei. The AVPV sends few projections to the caudal brainstem, but terminals were observed reliably in the periaqueductal gray and medial part of the nucleus of the solitary tract. Anterograde double-labeling experiments demonstrate terminals derived from neurons in the AVPV in close apposition to GnRH-containing neurons in the preoptic region, and to dopaminergic neurons in the arcuate nucleus. Thus, the organization of projections from the AVPV in female rats suggests that neurons in this nucleus may influence the secretion of luteinizing hormone and prolactin through direct projections to GnRH neurons and tuberoinfundibular dopaminergic neurons. PMID- 9214546 TI - Squamate Harderian gland: an overview. AB - BACKGROUND: The Harderian gland is an orbital feature found in most terrestrial vertebrates. Although there have been several reports on the structure of the squamate Harderian gland, there has been little recent discussion as to its potential function. This article reviews both the recent morphological observations and their implications on the potential functions of the squamate Harderian gland. METHODS: Literature on the gross structure, histochemistry, and ultrastructure of the squamate Harderian gland and associated structures was reviewed. These observations were then used to assess morphologically the likelihood of the proposed functions. RESULTS: A high level of morphological variation was found in the squamate Harderian gland. Three functional hypotheses, including roles in orbital lubrication, digestion, and vomerolfaction, were considered. Both morphology of the squamate Harderian gland and the presence of alternate secretory sources suggest that it is unlikely to function in orbital lubrication. There is little evidence to suggest a function in digestion. Both the presence of the connecting lacrimal apparatus and the reduced intrinsic secretory capacity of the vomeronasal organ suggest that the Harderian gland may function in vomerolfaction. CONCLUSIONS: The most likely role of the squamate Harderian gland seems to be in vomerolfaction. Morphological variations observed in the Harderian gland may mirror the different degrees and mechanisms of vomerolfaction. Further studies, including comparative morphological, experimental, and microchemical analyses, are required to test this hypothesis. PMID- 9214547 TI - Monoclonal antibody marker for olfactory sustentacular cell microvilli. AB - BACKGROUND: The olfactory epithelial sustentacular cells may support the survival and function of olfactory receptor neurons, but few reagents are available to mark and manipulate such cells. METHODS: Novel nasal cell-specific monoclonal antibodies were generated using whole cultured rat olfactory mucosal cells as the antigenic stimuli. They were characterized by immunostaining at the light level in rat tissues and newborn rat olfactory cell cultures, and at the electron microscopic level in adult tissues using freeze-substitution, post-embedding staining. RESULTS: An IgMkappa monoclonal antibody designated 1F4 selectively labeled apical surfaces of the rat olfactory and respiratory epithelia in tissue sections and what appeared to be sustentacular cells in olfactory cell cultures. Using electron microscopy, 1F4 bound selectively to the microvilli of sustentacular cells and ductal cells of Bowman's glands in the olfactory epithelium, and to the microvilli and cilia of ciliated but not secretory cells in the respiratory epithelium. No staining was detected in olfactory receptor neurons, basal cells, or two types of microvilli-bearing cells that differed from sustentacular cells. A contrasting antibody, 2H4, bound to granules of secretory respiratory cells. Developmental expression of 1F4 binding began at E17 and increased at and after E18/E19. Bulbectomy did not alter 1F4 immunoreactivity. Cell culture studies found that the 1F4 epitope was external and insensitive to trypsin treatment, and that both 1F4 and 2H4 positive cells required contact with aggregated cells for survival up to fifteen days in vitro. CONCLUSIONS: The antibody 1F4 is a useful marker and potential manipulation reagent specific for sustentacular cells and their microvilli. PMID- 9214548 TI - Hilum of the heart. AB - BACKGROUND: Organs with the juxtaposed entry or exit of their communications have the sites which should be termed hila. Despite juxtaposed communications of the heart, the name hilum cordis is absent in the Nomina Anatomica. RESULTS: This paper describes the hilum of the heart as the site bounded by the serous pericardium above heart base, ascending aorta and pulmonary trunk. CONCLUSION: Because the hilum of the heart is an equivalent to the hila of the other organs enveloped by the splanchnic mesoderm and thus far the term hilum cordis is absent in the Nomina Anatomica, the authors suggest to recognize a novel anatomical term hilum cordis. PMID- 9214549 TI - Fine structure of tuft cells of the main excretory duct epithelium in the rat submandibular gland. AB - BACKGROUND: Tuft cells, which are characterized by long microvilli with prominent rootlets and by vesicular and tubular profiles, are present in the mucosal epithelium of a number of hollow organs, including the main excretory duct of the rat submandibular gland. Despite their widespread occurrence, little is known of their function. METHODS: Main excretory ducts of the submandibular gland were obtained from 20 male Wistar rats and prepared for electron microscopic examination. Specimens also were subjected to various histochemical procedures, including HRP (horseradish peroxidase) uptake, the demonstration of catalase in peroxisomes, glycoconjugate cytochemistry, and ruthenium red staining. RESULTS: The features of the tuft cells in the MED (main excretory duct) basically were similar to those described in other organs. However, for the first time it was observed that in the submicrovillus zone and intermicrovillus space, there were many membrane-bound electron-dense granules, which resemble the so-called glycocalyceal bodies found on the apical surface of normal and neoplastic intestinal-type epithelial cells. Hypolemmal nerve terminals were seen to be in contact with the basal portion of tuft cells. After injection of HRP, reaction products were observed on the luminal surface of the microvilli, but not within the vesicles. Vesicles and tubules in the supranuclear cytoplasm exhibited a positive cytochemical reaction for glycoconjugates. The surface coat of tuft cell microvilli was stained with ruthenium red, but the vesicles and tubules in the supranuclear cytoplasm remained unstained. CONCLUSIONS: Our observations suggest that the tuft cells may engage in secretory activity in addition to reception. PMID- 9214550 TI - Development of tight junctions between odontoblasts in early dentinogenesis as revealed by freeze-fracture. AB - BACKGROUND: Mature odontoblasts possess junctional structures constituted by adherens, gap, and tight junctions. Although adherens and gap junctions appear early between odontoblasts, there is no information on the appearance and development of tight junctions between odontoblasts. In this study, we have examined freeze-fracture replicas of early dentinogenesis to study the development of tight junctions between odontoblasts and to determine whether these junctions are of zonular or macular type. METHODS: Upper first molar tooth germs of Wistar rats between 1 and 3 days old were fixed in buffered 4% glutaraldehyde/4% formaldehyde and subsequently cryoprotected with cacodylate buffered glycerol. Freeze-fracture replicas were obtained in a Balzers 301 apparatus, and early stages of dentinogenesis were examined in a Jeol 100 CX II electron microscope. RESULTS: In the stage of early dentine matrix prior to mineralization, odontoblasts exhibit only gap junctions. With the progression of development, the distal plasma membranes of odontoblasts show numerous short tight junctions formed by fused particles and grooves. In the stage of advanced mineralization, branched and continuous rows of fused particles or grooves constitute tight junctions of the focal or macular type. CONCLUSIONS: The present study shows that tight junctions of focal or macular type appear on distal plasma membrane of early odontoblasts during differentiation. Formation of tight junctions indicates the establishment of a distal membrane domain and maturation of odontoblasts. These events occur as mantle dentine formation ceases and circumpulpar dentine formation begins. PMID- 9214551 TI - Immuno-scanning electron microscope characterization of large tubules in human deciduous dentin. AB - BACKGROUND: This study was undertaken to elucidate the type and origin of collagen fibrils which construct the large tubules in deciduous coronal dentin by scanning electron microscope and anti-types I and III collagen antibody procedures. METHODS: The studies were performed on human deciduous teeth. The teeth were fixed in 4% paraformaldehyde solution and then fractured either mesio distally parallel to the long axis of the tooth or transversely perpendicular to the long axis of the tooth crown. The specimens were three-dimensionally observed employing the scanning electron microscope to distinguish the content of large tubules. Polyclonal antibodies of anti-type I and anti-type III collagen with 20 nm colloidal gold, and secondary electron imaging and backscatter electron imaging of high-resolution field emission scanning electron microscopy were used to examine the types of collagen fibrils. RESULTS: The large tubules extended from the vicinity of the incisal edge of the dentino-enamel junction to the pulp cavity. Inside the large tubules, fibers in compact bundles run parallel to the longitudinal axis of the tubules. The fiber bundles consisted of collagen fibrils which were 50-150 nm in diameter with typical cross striation. Immuno-scanning electron microscopy showed type I collagen-labelling gold particles and type III collagen-labelling gold particles to be abundant on the fibrils. Types I and III collagen-labelling gold particles were present on the banded collagen fibrils regardless of their diameter. CONCLUSIONS: It was found that type III collagen is present together with type I collagen on the fibrils constructing the large tubules of the human deciduous dentin. This immunohistochemical study suggested that the fibrils constructing the large tubules were derived from the von Korff fibers, and types I and III collagens formed copolymers. PMID- 9214552 TI - Quantitative study of the effects of long-term denervation on the extensor digitorum longus muscle of the rat. AB - BACKGROUND: In order to understand the cellular basis underlying the progressively poorer restorative capacity of long-term denervated muscle, we determined the effects of long-term denervation on the muscle fibers and satellite cell population of the rat extensor digitorum longus (EDL) muscle. METHODS: In 36 male rats, the right hind legs were denervated, and EDL muscles were removed 2, 4, 7, 12, and 18 months later. Muscles were either fixed for electron microscopic analysis or were dissociated into individual muscle fibers for direct fiber counting or for confocal microscopic analysis. RESULTS: The percentage of satellite cells rose from the 2.8% control value to 9.1% at 2 months of denervation; thereafter the percentage decreased to 1.1% at 18 months of denervation. The number of myonuclei per muscle fiber steadily declined from 410 in 4 month control muscle to 158 in 7 month denervated muscle. Up to 7 months of denervation, the total number of muscle fibers per muscle remained relatively constant at somewhat over 5,000. The calculated total satellite cell population in 4 month denervated EDL muscle was the same as that of controls at 65,000, but by 7 months of denervation it had declined to 21,000. With increasing time of denervation, the number of cross-sectional profiles of muscle fibers not containing nuclei rose from 14% in control muscle to 49% in 12 month denervated muscle. This was correlated with a pronounced regular clumping of the nuclei, with pronounced nonnucleated segments between nuclear clumps. CONCLUSIONS: Increasing times of denervation are accompanied by a pronounced decline in the number of myonuclei per muscle fiber and an initial rise and subsequent fall in satellite cell number. These changes are correlated with a decreasing restorative ability of these muscles over the same periods of denervation. Further work on the proliferative capacity of the remaining satellite cells is necessary before firm quantitative conclusions can be made. PMID- 9214553 TI - Electron microscopic study of long-term denervated rat skeletal muscle. AB - BACKGROUND: This study describes the ultrastructure of long-term denervated rat extensor digitorum longus and tibialis anterior muscles, with particular emphasis on understanding the cellular basis for the reduced restorative capacity of long term denervated muscle. METHODS: In 30 male WI/HicksCar rats, the right hindleg was denervated for periods of 1, 2, 4, 5.5, 6, 7, 12, 14, and 18 months before tissues were prepared for electron microscopy. RESULTS: Atrophy of muscle fibers was prominent by the second month post-denervation. At this time, type II fibers showed greater atrophy than type I fibers. At further periods of denervation, atrophy of all fibers was seen; and with increasing times of denervation the muscle fibers became surrounded by dense mats of collagen fibers. Muscle spindles persisted for the duration of this study. At two and four months, satellite cells showed signs of activation, such as elongated cytoplasmic processes and an increased concentration of cytoplasmic organelles. As denervation progressed, activated satellite cells became more widely separated from their associated muscle fibers, and basal lamina material was deposited between the satellite cells and muscle fibers. Some satellite cells broke free from their muscle fibers, and others acted as bridges between two muscle fibers. Evidence was seen of both muscle fiber degeneration and the regeneration of new muscle fibers, often more than one regenerating fiber beneath a single basal lamina. Loose folds of basal lamina were often present around atrophic muscle fibers. As denervation progressed, the morphology of individual muscle fibers varied. Some contained well-ordered lattice arrays of myofilaments, whereas in others considerable sarcomeric disorganization was evident. Mitochondria became smaller and rounded; elements of the sarcoplasmic reticulum proliferated and became more disorganized; lipid droplets, glycogen deposits, and autophagic vesicles were all present in the cytoplasm of atrophic muscle fibers. CONCLUSIONS: In addition to muscle fiber atrophy, long-term denervated muscles show evidence of myofiber and capillary death, as well as the deposition of massive amounts of interstitial collagen. These changes, all of which would appear to reduce the restorative capacity of the denervated muscle, take place concurrently with the morphological activation of satellite cells. The latter indicates that even in the denervated condition, restorative processes occur concurrently with regressive processes. PMID- 9214554 TI - Effect of hypoxia on the proliferation of retinal microvessel endothelial cells in culture. AB - BACKGROUND: To determine if hypoxia stimulates the proliferation of retinal microvessel endothelial cells in culture. METHODS: Bovine retinal microvessel endothelial cells were cultured in normoxic (95% air, 5% CO2) and hypoxic (2% O2, 5% CO2, 93% N2) conditions. Endothelial cells were identified by acetylated LDL and Factor VIII-related antigen immunocytochemical staining. Cells from passages three to eight were used in these experiments. Proliferation assays included cell counts by hemocytometer and autoradiographic analysis of incorporated 3H thymidine (3H-TdR). RESULTS: At day 4, cell counts of endothelial cells in hypoxia showed a 133% increase over those grown in normoxic conditions (N = 25, P < 0.01). Cell counts per day for 5 days were 121-181% greater in hypoxia. Autoradiography of endothelial cells exposed to 3H-TdR and counted every 12 hours for 60 hours exhibited labeling indices 112-118% higher in hypoxic conditions (P < 0.0001). Endothelial cells cultured under hypoxic conditions were smaller and spindle-shaped, whereas those grown under normoxic conditions were larger and more polygonal. CONCLUSIONS: Hypoxia increases DNA synthesis and stimulates proliferation of retinal microvessel endothelial cells in vitro and induces alterations in morphology. These results may be relevant to microvessel angiogenesis, which occurs in vivo under ischemic conditions. PMID- 9214555 TI - Acute action of choriogonadotropin on Leydig tumor cells: qualitative and quantitative transformation of lipid moieties. AB - BACKGROUND: Testicular Leydig cells use either exogenous or de novo synthesized cholesterol as the substrate for the production of testosterone with hormone stimulation. Although the long-term effect of trophic hormones on Leydig cell cholesterol uptake, storage, and deesterification has been well documented, the early effects of the human choriogonadotropin (hCG) on cell cholesterol/lipid distribution are not yet known. METHODS: Sections of cells treated with hCG for 15 sec to 30 min were examined by electron microscopy (EM) for the surface density of lipid moieties in the cytoplasm. In addition, the time-dependent distribution of lipids within the cytoplasmic inclusions and the ultimate destination of this substrate were evaluated by EM. The results were analyzed with standard morphometric methods. RESULTS AND CONCLUSION: The surface density of cytoplasmic lipid pools increased significantly within the 15 sec following the exposure of cells to hCG, and it tapered off to the control level in the subsequent 30 min. Such a fluctuation in the amount of cytoplasmic lipids may be due to (1) the quantity of released substrate from the reticular compartment or (2) the rate of its transport across the outer mitochondrial membrane to the inner mitochondrial cytochrome P450 side-chain cleavage, where steroidogenesis begins with the conversion of cholesterol to pregnenolone. These two processes were not quantitatively coordinated in the stimulated cell during the initial 30 min, resulting in a surplus of cytoplasmic lipid pools. To compensate for such uneven metabolic balance, the cell apparently disposed of the excess substrate by a mechanism of molecular regrouping from a micellar configuration to a bilayer structure followed by exocytosis. PMID- 9214556 TI - Distribution of leukocytes in the epithelium and interstitium of four regions of the Lewis rat epididymis. AB - BACKGROUND: Leukocytes expressing different surface markers were studied in four regions of the epididymis of Lewis rats. Cells resembling lymphocytes or monocytes had been described in the epididymis, but previous studies differed as to their nature and immunologic significance. METHODS: Frozen sections were immunocytochemically stained with monoclonal antibodies W3/25, OX-8, OX-42, and RLN-9D3, which are directed toward markers on CD4+ cells, CD8+ cells, macrophages, and B lymphocytes, respectively. The concentration of stained cells in the epithelium and interstitial tissue of the initial segment, caput, proximal cauda, and distal cauda regions was determined by a procedure based on the optical disector method. RESULTS: CD4+ leukocytes were present in greater concentration than CD8+ cells or macrophages in both the epithelium and interstitial tissue of all four regions. In the epithelium, the concentration of CD8+ leukocytes was greater than that of macrophages in the initial segment, caput, and distal cauda. In the interstitium, however, the concentration of macrophages exceeded that of CD8+ cells in both parts of the cauda. Macrophages and T lymphocytes were generally present in greater concentrations in the interstitium than in the epithelium, especially in the more proximal parts of the epididymis. In contrast to T cells, B lymphocytes were not detected in the interstitium or epithelium of any of part of the epididymis, despite prominent staining of B cells in other locations. CONCLUSIONS: The epididymal epithelium of the Lewis rat contains many T lymphocytes, which may correspond to 'halo' cells. CD4+ leukocytes predominate in all regions of the epididymis. The interstitium may function as a reservoir of leukocytes for the epithelial compartment. The epididymis is not normally a site for local immunoglobulin synthesis. PMID- 9214557 TI - Changes in the distribution of intermediate filaments in rat Sertoli cells during the seminiferous epithelium cycle and postnatal development. AB - BACKGROUND: Intermediate filaments (IFs) are components of the cytoskeleton. In mammalian Sertoli cell, IFs are formed by vimentin. Previous studies have shown some characteristics of its distribution in Sertoli cells, however, very little is known of its distributional changes during the seminiferous epithelium cycle and during postnatal development. METHODS: Immunohistochemical and electron microscopic methods were used to determine the distribution of vimentin-type IFs in rat Sertoli cells during the seminiferous epithelium cycle and postnatal development. RESULTS: The distribution of IFs in adult rat Sertoli cell showed distinct cyclic changes during the seminiferous epithelium cycle. At stages I-VI, bundles of IFs extend from the perinuclear region to the supranuclear and apical regions of the Sertoli cell. These apical extensions became shorter at stage VII, and at stages VIII-X IFs were observed only in the perinuclear region. Short apical extensions reappeared at stages XI-XII; and at stages XIII-XIV, they extended again into the apical region. During this cycle, IFs were always closely associated with the heads of elongate spermatids. IFs were also shown to be in close apposition to some specialized structures on the cell membrane, such as the ectoplasmic specialization between adjacent Sertoli cells. During postnatal (p.n.) development, IFs were mainly observed at the basal nuclear region on p.n. day 7. The IFs in the supranuclear or apical regions first appeared at p.n. day 14 and gradually increased during the development. The perinuclear IFs network was fully established by p.n. day 28 and the adult distribution pattern of the IFs was established by p.n. day 42. CONCLUSIONS: Vimentin-type IFs in rat Sertoli cells are a delicate endocellular network, which is centered in the perinuclear region and extends to the apical region of the cell. During the seminiferous epithelium cycle, the distribution of IFs changes in a stage-dependent manner and is closely related to the location of the heads of elongate spermatids. During postnatal development, IFs gradually increase in numbers and the main distribution area is transferred from the basal nuclear to the perinuclear and supranuclear regions. PMID- 9214558 TI - Localization of glycogen phosphorylase activity in liver of fasted normal and adrenalectomized rats and in fasted adrenalectomized rats after injection of dexamethasone. AB - BACKGROUND: The intralobular distribution of activity of glycogen phosphorylase (GP), a key enzyme in the breakdown of glycogen, was evaluated to determine changes during early glycogen synthesis. Hepatic GP activity was localized in normal and adrenalectomized (ADX) rats after fasting overnight and in fasted ADX rats stimulated to synthesize glycogen by administration of dexamethasone (DEX) 2 8 h prior to sacrifice. METHODS: Cryostat sections were incubated in medium containing appropriate substrate for demonstration of GP activity as indicated by glycogen synthesized by the enzyme during incubation. RESULTS: In sections from fasted normal rats, GP activity in hepatocytes varied from undetectable to substantial amounts with no notable periportal to pericentral gradient evident. In contrast, GP activity in sections from adrenalectomized fasted rats was concentrated in discrete aggregates in random hepatocytes throughout lobules. Two hours after DEX injection, GP enzyme activity occurred as single aggregates or in a dispersed pattern in many hepatocytes. By 4 h after DEX administration, most cells displayed GP enzyme activity, the concentration of which appeared to be greater in pericentral cells than in periportal cells. Eight hours after injection of DEX, GP enzyme activity had increased and appeared more evenly distributed throughout the lobules. CONCLUSIONS: These results suggest that GP activity became concentrated in limited regions of selected hepatocytes in fasted ADX rats. DEX stimulation of glycogen synthesis in these rats resulted in increased GP activity that was concentrated in pericentral cells after 4 h. After 8 h, activity increased and was more evenly distributed throughout the lobules. The increase in GP enzyme activity concurrent with overall glycogen synthesis suggests that the enzyme may participate in glycogen turnover. PMID- 9214559 TI - Larynx-associated lymphoid tissue (LALT) in young children. AB - BACKGROUND: Mucosa-associated lymphoid tissue (MALT) plays a central role in mucosal immunity. Whereas the characteristics and function of MALT in the intestine are well established, almost nothing is known about MALT in the larynx. METHODS: In this study we examined the morphology and the lymphocyte subset composition of MALT in the larynges of children who had died of sudden infant death or various defined traumatic or nontraumatic causes. RESULTS: Organized lymphoid tissue was found in the supraglottic parts of the larynx in nearly 80% of the children in both groups. This lymphoid tissue showed all morphological signs of MALT, such as typical lymphoid follicles with germinal centers, infiltration of the overlying epithelium by lymphocytes, and high endothelial venules (HEV). Thus we will use the term LALT (larynx-associated lymphoid tissue) to refer to this tissue. The lymphoid follicles of LALT contained mainly B lymphocytes with some CD4+ lymphocytes in the germinal centers. Remarkably, T lymphocytes of both subset types and B lymphocytes were observed in comparable numbers in the parafollicular area. CONCLUSIONS: We assume that LALT is a physiological structure of the larynx in young children. The morphology and the distribution of lymphocyte subsets are similar to those of MALT in the human gut. LALT may be a regular part of the mucosal immune system in young children with the role of respiratory inductive site for mucosal immunity. PMID- 9214560 TI - Developmental changes in the myocardial architecture of the chick. AB - BACKGROUND: Numerous studies describing myocardial architecture have been performed on the adult heart but considerably fewer have been made during embryonic or fetal development. To serve as a basis for interspecies comparison of ventricular morphology, and as a reference for studying the effects of experimental perturbations, we examined the development of chick throughout the entire incubation period. METHODS: Chick hearts from stage 14 (day 2) to stage 46 (day 21) were perfusion-fixed, and sectioned in transverse, frontal and sagittal planes. The ventricular myocardial architecture was examined and photographed in the scanning electron microscope. RESULTS: At embryonic stage 16 and earlier, the smooth-walled heart loop had an outer myocardial mantle, cardiac jelly, and endocardium. From stage 18, there was an outer compact and inner trabeculated myocardium. Trabeculated myocardium could be subdivided into the outer (basal) portion adjacent to the compact layer and the central (luminal) part. The outer basal layer could be distinguished from the inner luminal by shorter and finer trabeculae with small, round intertrabecular spaces. From stage 24, the patterns of trabeculae and intertrabecular spaces were ventricle-specific. Between stages 24 to 31, abundant trabeculations were present throughout both ventricular cavities. The trabeculae were initially radially arranged, but later adopted a spiral course, which persisted in a simplified form into adulthood. CONCLUSIONS: The ventricular myocardium undergoes distinctive morphogenesis, characterized by changes in trabecular patterning and orientation. We speculate that the embryonic trabecular architecture reflects the directions of the main stresses. Unlike fetal and adult hearts, which rely mostly on the compact myocardial layer, the trabeculae play a crucial role in the contractile function of the embryonic heart. PMID- 9214561 TI - Topography of mechanoreceptors in the shoulder joint region--a computer-aided 3D reconstruction in the laboratory mouse. AB - BACKGROUND: We investigated the pattern of distribution of corpuscular sensory nerve endings in the shoulder region of the laboratory mouse in relation to their functional properties. METHODS: Twelve adult female white NMRI-F2-mice were used. The topography of sensory nerve endings in the shoulder joint region was reconstructed by three-dimensional image processing by using serial silver stained sections of paraffin-embedded samples. Semithin sections obtained from additional samples were used for light microscopy. RESULTS: Within the fibrous layer of the joint capsule, three types of mechanoreceptors were identified: small lamellated corpuscles of the Pacini type, Ruffini corpuscles, and Golgi tendon organs. Intracapsular small lamellated corpuscles of the Pacini type (in an average number of 29/joint) were found mainly in three areas: in the predominantly flaccid tissue of the axillary region, in the denser ventromedial parts of the capsule, close to the scapula, and in the tight texture of the fiber bundles near the glenoid labrum. Ruffini corpuscles were identified only in small numbers (2/joint) in the ventral aspect of the articular capsule of two animals. Golgi tendon organs (14 or 15 receptors/joint) were discovered predominantly in close vicinity to the joint capsule at the muscle tendon junction of the inserting rotator cuff muscles and in the biceps brachii and triceps brachii muscles. CONCLUSIONS: In view of their location in the shoulder joint capsule and the glenoid labrum, corpuscular mechanoreceptors evidently play an important role in joint control by inducing protective reflex actions in phases of extreme or abnormal movement. The density of sensory receptors in distinct areas of the shoulder joint capsule appears to be related to zones that are subjected to increased biomechanical stress during physical activity. PMID- 9214562 TI - Renin and angiotensin II receptor expression in the brains of DES-treated Syrian hamsters. AB - BACKGROUND: The renin angiotensin system (RAS) promotes vasoconstriction. Expression of RAS is induced by different factors. METHODS: In this study, forebrain sections of hamster brains were studied by immunohistochemical methods to determine the location of renin-positive and angiotensin II receptor-positive cells. The brain sections were obtained from diethylstilbesterol- (DES-) treated hamsters, adult non-DES-treated hamsters, elderly non-DES-treated hamsters, neonatal hamsters, and 15-day fetal hamsters. Circulating renin activity was determined for all but the neonatal and 15-day fetal hamsters. RESULTS: Renin positive and angiotensin II receptor-positive vascular smooth muscle cells were observed in DES-treated hamsters. No positive cells were observed in neonatal, 15 day fetal, and adult non-DES-treated hamsters. Some expression was observed in elderly hamsters. CONCLUSIONS: Therefore, focal expression of the renin angiotensin system in brain vasculature was induced by the synthetic estrogen DES. PMID- 9214563 TI - Prenatal growth of the human vomeronasal organ. AB - BACKGROUND: Vomeronasal organs (VNOs) are paired epithelial structures located adjacent to the nasal septum that form in the late first trimester of human fetal development. Although VNOs have long been known to exist in fetal and adult humans, some studies continue to suggest that these structures may be degenerative or functionless. Little is known of the growth of the VNO. METHODS: The present study examined length and volume changes of the human VNO in 26 "normal" (10 female, 16 male) histologically prepared fetuses from the University of Pittsburgh and the University of Michigan across three trimesters (8-30 weeks postmenstrual age). A computer reconstruction technique was used to quantify lengths and volumes of right and left VNOs, and regression equations were generated to assess growth rates. RESULTS: A linear increase in VNO length and a logarithmic increase in VNO volume with increasing postmenstrual age was found. Volume increase was noted for both the vomeronasal epithelium and the lumen of the VNO. A comparison with most estimates of adult human VNO length suggested that further prenatal or postnatal size increase occurs. The growth curves also suggested a more rapid growth in VNO length and volume for females than for males. CONCLUSIONS: The present study demonstrates that the fetal human VNO commences volumetric increase in the early second trimester but does not achieve maximum size during fetal development. Further investigation is needed to determine whether the human VNO is sexually dimorphic in size. PMID- 9214564 TI - Distribution of primary motor nerve branches and terminal nerve entry points to the forearm muscles. AB - BACKGROUND: The information available on innervation pattern of the human forearm muscles in standard anatomy texts, although adequate for routine procedures, is not detailed enough for surgical reconstruction in complex injuries of the limb and for paralytic conditions of the forearm from peripheral nerve and spinal cord injuries. METHODS: The innervation pattern in 10 cadaveric forearms was studied. The contributions of the main nerve trunks to each forearm muscle was examined. The location and number of the primary motor nerve branching points and of the terminal nerve entry points to each muscle were investigated. The location of both the primary nerve branching points and terminal nerve entry points was presented as a percentage of forearm length measured from the lateral humeral epicondyle to the radial styloid. RESULTS: Seven of 19 forearm muscles were innervated from a single branch from the main nerve trunk. The remaining 12 received more than one primary branch. Two of 19 forearm muscles had only one terminal nerve entry point. The others had two or more each. In 13 of 19 forearm muscles, the statistical median location of the primary motor nerve branching points was within the proximal one-third of the forearm length and either more proximally or distally for the remainder. The statistical median location of the terminal nerve entry points was within the proximal one-third in 9 forearm muscles and within the middle one-third of the forearm in 8 forearm muscles. In two, it was located proximal to the elbow and in the distal one-third of the forearm, respectively. CONCLUSIONS: In lacerations across the forearm, where main nerve trunks are divided, mere repair of the nerve trunks would not address the denervation of muscle or segments of muscle by the division of the primary (or secondary) nerve branches traversing the wound and which took origin proximal to the laceration either from the divided nerve trunks or from other undamaged nerve trunks. Although the main nerve trunks may be intact, segmental crush injuries will defunction muscles by direct muscle damage or by damage to the terminal nerve entry points to the muscle. Knowledge of the location of the nerve branches and the terminal nerve entry points facilitates the insertion of electrodes at the motor points of forearm muscles for functional electrical stimulation in upper motor neuron lesions. The information in this study may also be usefully applied in selective denervation procedures to balance muscles in spastic upper limbs. PMID- 9214565 TI - Architecture of the human jaw-closing and jaw-opening muscles. AB - BACKGROUND: The human jaw-closing and jaw-opening muscles produce forces leading to the development of three-dimensional bite and chewing forces and to three dimensional movements of the jaw. The length of the sarcomeres is a major determinant for both force and velocity, and the maximal work, force, and shortening range each muscle is capable of producing are proportional to the architectural parameter volume, physiological cross-sectional area, and fiber length, respectively. In addition, the mechanical role the muscles play is strongly related to their three-dimensional position and orientation in the muscle-bone-joint system. The objective of this study was to compare relevant architectural characteristics for the jaw-closing and jaw-opening muscles and to provide a set of data that can be used in biomechanical modeling of the masticatory system. METHODS: In eight cadavers, sarcomere lengths, muscle masses, fiber lengths, pennation angles, and physiological cross-sectional areas were determined for the following muscles: superficial and deep masseter, anterior and posterior temporalis, anterior and posterior medial pterygoid, inferior and superior lateral pterygoid, posterior and anterior digastric, geniohyoid, posterior and anterior mylohyoid, and stylohyoid. To determine the spatial position of their action lines, the three-dimensional coordinates of the attachment sites were registered. RESULTS: Compared with the jaw openers, the jaw closers were characterized by shorter sarcomere lengths at the closed jaw, larger masses of contractile and tendinous tissue, larger physiological cross-sectional areas, larger pennation angles, shorter fiber lengths, shorter moment arms, and lower fiber-length-to-muscle-length ratios. In addition, architectural features differed across the muscles of the same functional group. Sarcomere length did not differ significantly among the regions of the same muscle. In contrast, in some muscles, significant intramuscular differences were found with respect to, e.g., physiological cross-sectional area, fiber length, pennation angle, and moment arm length. CONCLUSIONS: The results suggest that the jaw-closing muscles have architectural features that suit them for force production. Conversely, the jaw-opening muscles are better designed to produce velocity and displacement. PMID- 9214566 TI - Indirect way to estimate peak joint loads in life and in skeletal remains (insights from a new paradigm). AB - BACKGROUND: Bone adapts its strength and cross-sectional amount to the loads on it in now partly known ways. This makes it possible to estimate the unit loads on joint surfaces by an indirect method. METHODS: In essence, multiply the usual largest allowed compression load on a unit cross section area of epiphyseal trabecular bone (now approximately known), by the cross sectional amount of that bone that supports a unit area of the joint surface (partly known and readily measured by histomorphometry). This would equal the usual largest compression unit load on both the joint surface and the articular cartilage supported by that trabecular bone. RESULTS: Suggested typical peak unit loads on synovial joint surfaces in different joints and/or parts of joints could range from approximately 2 up to approximately 50 megapascals. CONCLUSIONS: Besides its use in studies of joint development, physiology and osteoarthritis in living subjects, this method could estimate muscle strength and joint loads from skeletal remains in anatomical, anthropologic, forensic-pathological, and even paleontologic studies. PMID- 9214567 TI - Outbreak of leptospirosis among white-water rafters--Costa Rica, 1996. AB - On October 15, 1996, a physician notified the Illinois Department of Public Health about five patients with an unknown febrile illness who had returned from a white-water rafting trip on flooded rivers in Costa Rica during September 27 28, 1996. The five patients had been members of a white-water rafting expedition involving 26 rafters from five states, the District of Columbia, and Costa Rica. This report summarizes the findings of the multistate investigation conducted by the Illinois Department of Public Health and by CDC in collaboration with the Ministry of Health of Costa Rica. The findings implicated leptospirosis as the cause of disease and contaminated river water as the probable source of illness. PMID- 9214568 TI - Progress toward global eradication of poliomyelitis, 1996. AB - Substantial progress was achieved during 1996 to further implement the World Health Organization (WHO)-recommended strategies for the global eradication of poliomyelitis (1). An international coalition of partners supporting the eradication effort in countries with endemic polio includes WHO, Rotary International, CDC, United Nations Children's Fund, and governments of countries with and without endemic polio. This report updates progress toward global polio eradication based on information available at WHO as of April 30, 1997. PMID- 9214569 TI - Prevalence of cardiovascular disease risk-factor clustering among persons aged > or = 45 years--Louisiana, 1991-1995. AB - Cardiovascular disease (CVD), including coronary heart disease, stroke, and hypertensive disease, is the leading cause of death in Louisiana and in the United States and, in 1994, accounted for 43.7% and 45.2% of all deaths among persons aged > or = 45 years in Louisiana and in the United States, respectively. The primary risk factors for CVD are hypertension, high cholesterol, diabetes, overweight, cigarette smoking, and physical inactivity. The first four of these risk factors may cluster in some persons and have been identified as components of a syndrome known as metabolic cardiovascular syndrome (1) or the "deadly quartet" (2). This syndrome is characterized by a persistent state of insulin resistance and compensatory hyperinsulinemia that may be etiologically related to the four risk factors. Persons with one of the four risk factors are at increased risk for having any of the other three (3). To determine the prevalence of risk factor clustering among older residents of Louisiana, CDC analyzed data from the 1991, 1992, 1993, and 1995 Louisiana Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the results of this analysis, which documented a prevalence of clustering of the four risk factors among 1.7% of the respondents aged > or = 45 years. PMID- 9214570 TI - Increased levels of 4-hydroxynonenal modified proteins in plasma of children with autoimmune diseases. AB - Analysis of serum samples of healthy children (n = 11) and children with Systemic Lupus Erythematosus (SLE), (n = 21) was performed by SDS-PAGE and immunoblot with an antibody directed against proteins modified by lipid peroxidation (LPO) product 4-hydroxynonenal (HNE). A single major stained protein band was detected. By comparison of the molecular weights in nonreducing and reducing SDS-PAGE was found that the main protein modified by HNE is immunoglobulin G. Significantly higher concentrations of the aldehyde modified protein were found in children with high disease activity of SLE measured by SLE disease activity index (SLEDAI). Lipid peroxidation measured by malondialdehyde and 4-hydroxynonenal concentrations show an enhanced level of both compounds also in patients with the active autoimmune disease. Therefore, it can be assumed that free radical mediated processes play a pathophysiological role in the active phase of SLE and HNE-modified serum proteins are a further parameter for the detection of in vivo LPO. PMID- 9214571 TI - Protein carbonyl measurement by a sensitive ELISA method. AB - We describe a new immunoassay for measuring protein carbonyls as an index of oxidative injury. Protein samples were reacted with dinitrophenylhydrazine then adsorbed to wells of an ELISA plate before probing with a commercial antibody raised against protein-conjugated dinitrophenylhydrazine. The biotin-conjugated primary antibody was then reacted with streptavidin-biotinylated horseradish peroxidase for quantification. The method was calibrated using oxidized albumin and results correlated well with the colorimetric carbonyl assay. The method required only 60 microg protein and was used to analyze the amount of protein carbonyls in plasma and lung aspirate samples. It was sensitive in the 0-2.5 nmol/mg protein range within which clinical samples fell and was linear up to 10 nmol/mg protein. The ELISA method for protein carbonyls is more sensitive and discriminatory than the colorimetric assay and should have wide application for analysing experimental and clinical samples, especially where concentrations are low and where only small amounts of sample are available. PMID- 9214572 TI - Reaction of melatonin and related indoles with hydroxyl radicals: EPR and spin trapping investigations. AB - It has been suggested that the indole hormone melatonin (N-acetyl-5 methoxytryptamine, MLT) is an important natural antioxidant and free radical scavenger [J. Pineal Res., 14:51; 1993]. In the present work we determined the rate constants, k(r), for scavenging .OH radicals by melatonin, 5 methoxytryptamine (5-MeO-T), 5-hydroxytryptamine (serotonin, 5-OH-T), 6 chloromelatonin (6-Cl-MLT), 6-hydroxymelatonin (6-OH-MLT), and kynurenine (KN) in aqueous solutions. Hydroxyl radicals were generated using a Fenton reaction in the presence of the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO), which competed with the indoles for the radicals. It was found that MLT reacts with .OH with k(r) = 2.7 x 10(10) M(-1) s(-1). Other indoles and KN reacted with .OH radicals with similarly high rates (k(r) > 10(10) M(-1) s(-1)). In contrast to nonhydroxylated indoles (MLT, 6-Cl-MLT, and 5-MeO-T), hydroxylated indoles (5-OH T and 6-OH-MLT) may function both as .OH promoters and .OH scavengers. The melatonin precursor serotonin promoted the generation of .OH radicals in the presence of ferric iron and H2O2, and the melatonin metabolite 6-hydroxymelatonin generated large quantities of .OH radicals in aerated solutions containing Fe3+ ion, even in the absence of externally added hydrogen peroxide. These reactions may be relevant to the biological action of these physiologically important indolic compounds. PMID- 9214573 TI - Sensitivity of protein sulfhydryl repair enzymes to oxidative stress. AB - According to their demonstrated activities, the thiol-disulfide oxidoreductase (TDOR) enzyme systems [thioltransferase (glutaredoxin) and GSSG reductase; and thioredoxin and thioredoxin reductase] are expected to provide the primary cellular mechanism for protection and repair of sulfhydryl proteins under oxidative stress. Since all four enzymes have active site dithiol moieties, they may be vulnerable to oxidative damage themselves. Therefore, an hydroxyl radical generating system (chelated ferrous iron in combination with hydrogen peroxide) was used to document the relative sensitivity of each of the enzymes to oxidative stress in vitro. At particular concentrations of enzymes and oxidant system, all of the enzymes were deactivated nearly completely, but different patterns of susceptibility were observed. At the approximate physiological concentration of each enzyme thioredoxin and thiol-transferase were largely deactivated with 1 mM Fe2+-ADP, 1 mM H2O2; whereas thioredoxin reductase and GSSG reductase were much less sensitive: 10 microM thioredoxin (88% deactivated), 1 microM thioltransferase (72%), 2 microM thioredoxin reductase (5%), and 0.1 microM GSSG reductase (17%). As the concentration of the oxidant system was decreased stepwise from 1 mM to 1 microM to mimic conditions that may be associated with oxidative tissue injury in situ, deactivation of thioredoxin was decreased proportionately, whereas thioltransferase remained much more susceptible. As expected GSH and other radical scavengers protected thioltransferase from deactivation by Fe(ADP)-H2O2. To test the susceptibility of the TDOR enzymes to oxidative stress in a physiological-like setting, isolated perfused rabbit hearts were subjected to 30 min ischemia and 30 min reperfusion. The GSH/GSSG ratio and total dethiolase activity (thioltransferase and thioredoxin systems) remained unchanged relative to control hearts, indicating that overall redox status and sulfhydryl repair activity are maintained during moderate oxidative stress in situ. PMID- 9214574 TI - In vivo exposure to ozone depletes vitamins C and E and induces lipid peroxidation in epidermal layers of murine skin. AB - To evaluate skin susceptibility to ozone (O3) and to localize possible oxidative damage within the skin layers, hairless mice were exposed to 10 ppm O3 or air (0 ppm O3) for 2 h. The mice were euthanized, the skin removed and frozen. Three skin layers (upper epidermis, lower epidermis/papillary dermis, and dermis) were separated, antioxidant concentrations (alpha-tocopherol and ascorbic acid) and the lipid peroxidation product malondialdehyde (MDA) measured. In the upper epidermis, O3 significantly depleted alpha-tocopherol (22%; p < .05) and ascorbic acid (55%; p < .01). These antioxidants were unchanged by O3 in the lower skin layers. More remarkably, MDA increased ten-fold in the upper epidermis (p < .001) and two-fold in the lower epidermis/papillary epidermis (p < .05); it was unchanged in the dermis. Thus, exposure to O3 in vivo depletes ascorbic acid and alpha-tocopherol and strongly induces lipid peroxidation in skin. High MDA concentrations measured in the upper epidermis suggest that O3 reacts directly with fatty acids on the skin surface layers. These results further suggest that chronic exposure to lower O3 concentrations found in urban smog could potentially have implications for skin health. PMID- 9214575 TI - Injury to cultured liver endothelial cells after cold preservation: mediation by reactive oxygen species that are released independently of the known trigger hypoxia/reoxygenation. AB - When cultured liver endothelial cells were incubated in cold University of Wisconsin (UW) solution under hypoxic conditions, 3 +/- 2% of cells had lost viability after 25 h. Simulating reperfusion by returning the cells to normal cell culture conditions increased the injury to 30 +/- 11% after 3 h of recultivation. An injury of similar time course was observed after cold normoxic incubation in UW solution when the loss of viability increased from 20 +/- 5% after the cold incubation to 60 +/- 8% during a 3 h recultivation period. The loss of viability during recultivation was decreased by hypoxia, by the addition of 5,5-dimethyl-1-pyrroline N-oxide or of dimethyl sulfoxide to the cell culture medium, or by preincubating the cells with deferoxamine. The injury was accompanied by lipid peroxidation and was dependent on the period of cold incubation in UW solution. These results suggest the occurrence of a cold preservation-induced "recultivation injury" mediated by reactive oxygen species. This cold-preservation-induced injury--occurring independently of hypoxia/reoxygenation--is likely to constitute an additional component of reperfusion injury after cold preservation of the liver. PMID- 9214576 TI - Hypochlorous acid disrupts the adhesive properties of subendothelial matrix. AB - We have investigated whether the cell adhesion-promoting properties of the subendothelial matrix are affected by exposure to neutrophil-derived oxidants. Native subendothelial matrix was exposed to increasing doses of H2O2 in the presence of myeloperoxidase and Cl- or to reagent hypochlorous acid (HOCl). Increasing doses of either oxidant system resulted in progressive loss in the adhesive properties of the matrix, and phase contrast microscopy showed that the cells failed to attach to and spread on the oxidant-treated surface. When cells were replated on the treated matrix in the presence of 20% serum, they did attach, but showed abnormal spreading and morphology in longer-term culture. In a modified ELISA system, binding of antibodies specific to fibronectin, thrombospondin and laminin was also disrupted by prior exposure of the matrix to HOCl. Of these components, the cell-binding region of fibronectin was most affected by HOCl, thrombospondin and laminin were less sensitive, and the collagen-binding region of fibronectin was the most resistant. SDS-PAGE of 35S labelled subendothelial matrix proteins indicated that there was no major irreversible crosslink formation or fragmentation after exposure to HOCl or the myeloperoxidase system, although formation of disulfides is quite likely. PMID- 9214577 TI - Psychomotor effects of dopamine infusion under decreased glutathione conditions. AB - Administration of buthionine sulfoximine (BSO) selectively inhibits glutathione (GSH) biosynthesis, thereby inducing a GSH deficiency. Because GSH plays a critical role in intracellular antioxidant defense, decreased GSH levels in the brain may result in less oxidative stress (OS) protection. Thus, the pro-oxidant effects of dopamine (DA), which rapidly oxidizes to form reactive oxygen species, may increase. In this study, the behavioral consequences of reduced OS protection were examined by administering BSO (3.2 mg in 30 microl Ringer's solution, intracerebroventricularly) every other day for 12 d to male Fischer 344 rats. In addition, DA (15 microl of 500 microM) was administered every day; when given on the same day as BSO, it was either 1 h after BSO (BSO + DA group) or 1 h before BSO (DA + BSO group). Tests of psychomotor behavior--rod walking, wire suspension, and plank walking--were performed five times during the experiment. BSO + DA administration, but not DA + BSO, impaired performance by decreasing latency to fall in the rod and plank walk tests compared to a vehicle only (Ringer's) group. Therefore, depletion of GSH with BSO, followed by DA treatment, produced deficits in psychomotor behavior. These deficits are similar to those seen in aged rats, suggesting that the oxidation of DA coupled with a reduced capacity to respond to OS may be responsible for the induction of age-related motor behavioral deficits. PMID- 9214578 TI - Effect of oxidative stress on membrane structure: small-angle X-ray diffraction analysis. AB - Free radical damage to cellular membranes appears to underlie alterations in function in aging and various pathological processes, including cardiovascular disease. The objective of this study was to directly characterize changes in the molecular structure of membrane lipid bilayers resulting from oxidative stress. Membrane samples reconstituted from either synthetic or cardiac phospholipids enriched with polyunsaturated fatty acids were examined at high resolution using small-angle x-ray diffraction methods. In this study, Fe2+/ascorbate-induced lipid peroxidation produced significant and dose-dependent alterations in the basic physical structure of the phospholipid bilayer. Electron density profiles (A vs. electrons/A3) calculated from the x-ray diffraction data showed a marked reduction in the hydrocarbon core width of dilinoleoyl phosphatidylcholine (DLPC) bilayers from 36 A to 32 A, and a decrease in overall membrane width, including surface hydration, from 48.7 A to 44.6 A. In addition, a broad decrease in molecular volume was observed +/-3-10 A from the center of the membrane bilayer, along with interdigitation of the terminal methyl segments. Pronounced changes in the lipid bilayer structure following oxidative stress were also observed in membranes reconstituted from cardiac lipids, including a 4 A reduction in hydrocarbon core width from 40 A to 36 A and interdigitation of the terminal methyl segments. These data provide direct evidence for changes in membrane hydrocarbon core width and molecular volume resulting from phospholipid peroxidation, which may contribute to perturbations in membrane structure/function relationships associated with aging and cardiovascular disease. PMID- 9214579 TI - The importance of sodium pyruvate in assessing damage produced by hydrogen peroxide. AB - Instability of hydrogen peroxide solutions was noted during the experimental exposure of human cells in culture to hydrogen peroxide in experiments designed to study the production and repair of DNA single-strand breaks. A hydrogen peroxide concentrate was diluted into culture medium, which was then added to experimental cell cultures at various times, with all cultures assessed for DNA damage at 2 h. Only cells treated by the first addition had observable DNA damage. This result was unexpected since these cells had had the maximum repair time. It was determined that the hydrogen peroxide had been eliminated by the culture medium. To determine the mechanism of this elimination, 200 microM hydrogen peroxide was added to various cell culture components, and the solutions were assayed for hydrogen peroxide after 1 h at 37 degrees C. Although most components (except the balanced salts) showed some hydrogen peroxide degradation, it was found that sodium pyruvate was most effective, by a wide margin, in eliminating hydrogen peroxide and its toxic effects. This was confirmed by addition of pyruvate to balanced salt solutions or buffers, and observing the same elimination of hydrogen peroxide. We subsequently found a few earlier reports describing the decarboxylation reaction between hydrogen peroxide and pyruvate, but no kinetic measurements have been published and there seems to be no general appreciation for the very high efficiency of this reaction. The present work presents a preliminary assessment of the importance of pyruvate in the study of hydrogen peroxide and other reactive oxygen species in mammalian cell culture. PMID- 9214580 TI - Expression of CuZn- and Mn-superoxide dismutase in human colorectal neoplasms. AB - Decreased intracellular SOD protein levels and activity have been related with malignancy in the past. To investigate their relevance in the carcinogenetic process in the colon, we determined quantitatively CuZn-SOD and Mn-SOD levels and total SOD activity by histochemical means in human normal colorectal mucosa, adenomas, and carcinomas. Protein levels and activity were significantly decreased in carcinomas. CuZn-SOD protein levels, but not Mn-SOD levels or total SOD activity were related with differentiation grade and to a lesser extent with Dukes stage. Moderately differentiated carcinomas and Dukes stage A carcinomas showed lowest levels. Some carcinomas expressed elevated levels of CuZn-SOD and this was an indication of poor survival. It is concluded that decreased SOD expression is not a prognostic marker and seems to be a secondary phenomenon rather than directly linked with the carcinogenetic process. PMID- 9214581 TI - Neutrophil antioxidant capacity during the respiratory burst: loss of glutathione induced by chloramines. AB - Low-molecular weight antioxidants in rat peritoneal neutrophils undergo rapid redox recycling, so measurements were made of their initial content and subsequent changes during the respiratory burst, when superoxide formation is maximized. Endogenous vitamin E, ascorbate and total glutathione (reduced + oxidized) were not significantly changed during 30 min of respiratory burst, which was stimulated by phorbol 12-myristate 13-acetate (PMA). When de novo synthesis of glutathione was inhibited by buthionine-[S,R] sulfoximine (BSO), the glutathione content rapidly decreased in activated neutrophils but not in resting cells. The lost total glutathione was recovered neither from the incubation medium nor as a protein-bound form, which suggests that irreversible oxidation of glutathione occurs. Furthermore, the glutathione loss continues even 30 min after PMA stimulation, when the respiratory burst has almost ceased. The decrease of glutathione was prevented by added catalase, or by addition of NaN3 or KCN which inhibits myeloperoxidase (MPO). Superoxide dismutase had no protective effects. These findings suggest the involvement of an MPO-H2O2-halide system in the accelerated consumption of glutathione during the respiratory burst. Additional studies showed that neutrophil-derived chloramines found in the extracellular medium could lead to intracellular glutathione loss. Incubation of resting cells with chemically produced membrane permeable monochloramine in the presence of BSO resulted in a decrease of glutathione, whereas membrane-impermeable taurine chloramine was less effective. We conclude that chloramines are responsible for accelerated glutathione turnover in neutrophils during the respiratory burst. Permeable extracellular chloramines derived from the respiratory burst activity, such as monochloramine, can reenter cells and react with thiols. PMID- 9214582 TI - Modulating role of endogenous reduced glutathione in tert-butyl hydroperoxide induced cell injury in isolated rat hepatocytes. AB - The role of endogenous reduced glutathione (GSH) in tert-butyl hydroperoxide (TBHP)-induced cell injury was examined in isolated rat hepatocytes. When liver cell injury was estimated from release of transaminases from hepatocytes into the incubation medium, cell injury in hepatocytes (2 x 10(6) cells/ml) incubated in Hanks' balanced salt solution (pH 7.2) containing 1.0 mM TBHP at 37 degrees C was potentiated with enhanced lipid peroxidation by prior depletion of intracellular GSH which was induced by diethylmaleate, a GSH depletor. GSH-depleted hepatocytes were incubated with gamma-glutamylcysteinylethyl ester (gamma-ECOEt), which is known to be converted to GSH via glutathione synthetase after its hydrolysis by esterase, at concentrations of 1.0 to 10 mM in order to replenish intracellular GSH. Although TBHP-induced cell injury and lipid peroxidation were enhanced in GSH-depleted hepatocytes, these enhancements were prevented with the consumption of intracellular GSH in GSH-depleted hepatocytes pretreated with 5.0 mM gamma ECOEt. These preventive effects were observed at any time point during the TBHP treatment over a 60 min period and depended on the concentration of gamma-ECOEt used. But, no preventive effect was found in GSH-depleted hepatocytes pretreated with 5.0 mM GSH. No prevention of the potentiation of TBHP-induced cell injury found in GSH-depleted hepatocytes occurred in GSH-depleted hepatocytes pretreated with both 5.0 mM gamma-ECOEt and 250 microM bis-(p-nitrophenyl)phosphate, a nonspecific esterase inhibitor. gamma-ECOEt treatment caused an increase in intracellular GSH content in GSH-depleted hepatocytes, while treatments of both gamma-ECOEt and the esterase inhibitor caused no increase in intracellular GSH content in the cells. These results indicate that endogenous GSH modulates TBHP induced cell injury and lipid peroxidation in isolated rat hepatocytes. The present results suggest that endogenous GSH should play a critical role in TBHP induced cell injury in isolated rat hepatocytes and that in rat hepatocytes treated with TBHP, enhanced lipid peroxidation with the consumption of intracellular GSH could be associated with the initiation of cell injury. PMID- 9214583 TI - Decrease in accessible thiols as an index of oxidative damage to membrane proteins. AB - The effect of several oxidative agents (hydrogen peroxide, tert-butyl hydroperoxide, menadione, AAPH, peroxynitrite and ionizing radiation) on the ratio of weakly to strongly immobilized residues of erythrocyte membrane-bound maleimide-tempo spin label (h(w)/h(s) ratio) was studied in order to test the hypothesis that a decrease in the h(w)/h(s) ratio may be a general index of oxidative damage to membrane proteins. Most of the agents studied decreased though H2O2 almost did not affect and ionizing radiation increased the h(w)/h(s) ratio. In parallel, the ratio of DTNB accessible/DTNB inaccessible membrane protein-SH groups was determined from membrane-SH group measurements with the Ellman reagent in the absence and in the presence of sodium dodecyl sulfate. This ratio decreased in all cases studied and seems to be a more universal and easy to measure parameter to describe the oxidative damage to membrane proteins. PMID- 9214584 TI - Oxidative stress and defense mechanisms in plants: introduction. PMID- 9214585 TI - Use of transgenic plants to study antioxidant defenses. AB - The abundance of O2 and the highly energetic electron transfer reactions associated with thylakoid membranes make chloroplasts a major source of reactive oxygen intermediates (ROI) in photosynthetic tissues of plants. Attempts to reduce oxidative damage in chloroplasts have included the manipulation of ROI scavenging enzymes by gene transfer technology. Much of this work has focused on chloroplast-localized superoxide dismutase (SOD), both chloroplast-targeted and cytosolic ascorbate peroxidase (APX) and glutathione reductase (GR). Increased activity of SOD in chloroplasts of transgenic tobacco plants generally leads to increased protection from membrane damage caused by exposure to methyl viologen (MV). In addition, overexpression of chloroplastic Cu/Zn SOD can lead to increased protection from photooxidative damage caused by high light intensity and low temperatures. Transgenic tobacco plants that overexpress APX either in the cytosol or chloroplastic compartments also show reduced damage following either MV exposure or photooxidative treatment and transgenic plants that express increased levels of GR have elevated pools of ascorbate and GSH. Though still preliminary, these results clearly indicate that alterations in the expression of enzymes involved in ROI-scavenging can have significant metabolic effects and provide the hope that this strategy can be used to develop crop plants with increased stress tolerance. PMID- 9214586 TI - The effects of ozone on antioxidant responses in plants. AB - The response of plants to ozone exposure includes a number of physiological and biochemical changes that are the direct result of selective increases or decreases in gene expression and the resulting changes in the accumulation of the corresponding protein products. Major classes of ozone-induced proteins include antioxidant enzymes and a number of stress-related proteins associated with other biotic and abiotic stresses. In particular, there is a significant overlap in the pattern of gene induction observed in ozone-treated plants and plants exhibiting pathogen defense responses. Current knowledge concerning the specific molecular events associated with the alterations of gene expression caused by ozone and the precise roles of ozone-induced proteins in conferring tolerance to ozone is rather limited. This review summarizes some of the recent results that have been obtained concerning the molecular basis of ozone-induced responses in plants, with an emphasis on studies of the model plant system, Arabidopsis thaliana. These studies demonstrate that ozone-induced responses are caused in part by the activation of a salicylic acid dependent signaling pathway that is also required for the expression of resistance to microbial pathogens. PMID- 9214587 TI - Regulation of catalases in Arabidopsis. AB - The catalase multi-gene family in Arabidopsis includes three genes encoding individual subunits which associate to form at least six isozymes that are readily resolved by non-denaturing gel electrophoresis. CAT1 and CAT3 map to chromosome 1, and CAT2 maps to chromosome 4. The nucleotide and deduced amino acids sequences of the three coding regions are highly related to each other and to other catalases. Both the individual isozymes and the individual subunit mRNAs show distinct patterns of spatial (organ-specific) expression. Six isozymes are detected in flowers and leaves and two are seen in roots. All three mRNAs are highly expressed in inflorescences, and CAT2 and CAT3 are highly expressed in leaves. All three mRNAs are detectable in freshly imbibed seeds, although the pattern of mRNA relative abundance varies among the three genes during early germination. CAT1 and CAT2 mRNA abundance is induced by light. In contrast, CAT3 is negatively light-responsive. CAT2 and CAT3 mRNA abundance is controlled by the circadian clock. Interestingly, the peak in CAT3 mRNA abundance occurs in the subjective evening, which is out of phase with expression of the Arabidopsis CAT2 catalase gene that shows clock-regulated expression gated to the subjective early morning. CAT1 mRNA abundance is not clock-regulated. PMID- 9214588 TI - Response of the maize catalases to light. AB - The three maize catalase genes respond differentially to light signals. Expression of Cat1 is light independent while expression of Cat2 and Cat3 is light responsive. Upon exposure to light there is rapid accumulation of CAT-2 protein in leaves, due to both increased transcript accumulation and increased translation of the Cat2 message. Short UV light pulses also cause a strong transient induction of Cat2 gene expression, while long term exposure to UV does not affect the rate of Cat2 transcription. The Cat3 gene of maize exhibits a transcriptionally regulated circadian rhythm. The induction of the Cat3 circadian expression in etiolated leaves is probably regulated by a very low fluence phytochrome response; the involvement of a blue light/UV-A and a UV-B photoreceptor is also possible. Regulatory elements located on the Cat3 promoter have recently been identified and their significance in the complex light response of the gene is being investigated. Possible physiological role(s) of the light responding maize catalases Cat2 and Cat3 are discussed. PMID- 9214589 TI - Influence of UV-light on the expression of the Cat2 and Cat3 catalase genes in maize. AB - The effects of UV (ultraviolet) -irradiation on the expression of the antioxidant genes Cat2 and Cat3, encoding the CAT-2 and CAT-3 catalases in maize were examined. Cat2 and Cat3 transcript accumulation was analyzed in leaves of maize seedlings grown under different light conditions, and subsequently exposed to UV light. Under DD-(constant darkness) and LL- (continuous light) conditions, as well as under a 12h D/L- (dark/light) photoperiod, the Cat2 mRNA was expressed at low and constant levels. In contrast, Cat3 transcript accumulation was constant and about 10 times higher than that of Cat2 under DD or LL, while the expression of Cat3 exhibits a typical circadian rhythm under a 12h D/L photoperiod. UV- light pulses in the range of 290 to 400 nm strongly induce the expression of Cat2. Upon removing the UV-B portion (290-310 nm) of the UV-spectrum the maximal Cat2 transcript level was reduced by about 60%. On applying UV-light of the same quality in addition to visible light, the expression of Cat2 was induced. DNA damage caused by UV-light and induction mediated by a UV-light photosensory system are suggested. Further, it is suggested that the Cat3 circadian rhythm may be regulated by a blue light/UV-A and a UV-B photoreceptor. If DD and LL grown plants that exhibit no circadian oscillation, were exposed to constant UV-light in the range of 290 to 400 nm a circadian rhythm was induced; indicating that UV light may function as an additional environmental cue to entrain the Cat3 circadian rhythm. Since, Cat2 and Cat3 showed distinct responses to UV light, it is suggested that both genes may act to scavenge reactive oxygen species (ROS) generated by UV-light to protect the plant from oxidative damage. PMID- 9214590 TI - The regulation and function of tobacco superoxide dismutases. AB - Our knowledge of superoxide dismutase (SODs) in tobacco has increased greatly during the past few years. Genes encoding the four identified SOD isoforms of tobacco have been isolated and characterized. Analysis of promoter-beta glucuronidase fusions has provided information on the cellular expression of SODs in tobacco and has constituted the basis for studying SOD regulation. Constitutive overproduction of SOD has been shown to confer increased tolerance to stress and has started to reveal subtle biochemical differences between SOD isoforms. Thus, thanks to its convenience for molecular and physiological studies, tobacco has come forth in recent years as an excellent model system for studying the regulation and function of SOD in dicotyledonous plants. PMID- 9214591 TI - 4-hydroxynonenal-induced relaxation of human mesenteric arteries. AB - The effect of 4-hydroxynonenal (4-HNE), a circulating lipid peroxidation product, on the vascular tone of human mesenteric arteries is studied. 4-HNE promotes relaxation of human mesenteric arterial rings in a concentration-dependent manner. Removal of the endothelium or treatment with N(G)-nitro-L-arginine methyl ester hydrochloride (L-NAME; 10(-4) M) partially prevented 4-HNE-induced relaxation, thus suggesting the intervention of nitric oxide from endothelial origin in the vascular effects of 4-HNE. PMID- 9214592 TI - Increased transcription of the astrocyte gene GFAP during middle-age is attenuated by food restriction: implications for the role of oxidative stress. AB - Glial fibrillary acidic protein (GFAP), an intermediate filament of astrocytes, shows increased expression during aging. Because we found that chronic food restriction retards the increase of GFAP mRNA in aging rats and because food restriction decreases the load of oxidized proteins and lipids in association with increased life span, we investigated the regulation of GFAP during oxidative stress and aging. First, we showed that food restriction decreased the transcription of GFAP in aging rats. This result generalizes effects of food restriction on age changes of transcription; whether transcription decreases during aging as in hepatic genes, or increases during aging as in astrocytic GFAP, food restriction attenuates the age change. Moreover, food restriction decreased microglial activation during aging, which suggested the hypothesis that GFAP expression is sensitive to oxidative stress. Because GFAP transcription in cultured glia is increased by oxidative stress in response to hydrogen peroxide and cysteamine whether or not microglia were present, we conclude that responses of GFAP to oxidative stress in astrocytes do not depend on microglial activation. The results implicate oxidative stress in the increased expression of GFAP during aging, but also in responses to brain injury. PMID- 9214593 TI - Spectra of spontaneous mutations at the hprt locus in colorectal carcinoma cell lines defective in mismatch repair. AB - Spectra of spontaneous mutations at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus in colon carcinoma cell lines HCT116 and HCT-15 deficient in mismatch repair and displaying mutator phenotypes were determined. HCT116 and HCT-15 cells, respectively, harbour a mutation in the mismatch repair gene hMLH1 and GTBP. The mutation frequency at the hprt locus in both cell lines was elevated by about two orders, but the microsatellite instability in HCT116 cells was one order higher than in HCT-15 cells. Except for one mutant of HCT-15, all the mutations (114/115) were point mutations; base substitutions of various types and frameshifts (deletions/insertions of less than a few bases, predominantly of +/-1 bp). Base substitutions (57%) and frameshifts (43%) occurred at a comparable rate in HCT116, whereas base substitutions (92%) were the major mutational events in HCT-15. Most frameshifts in HCT116 occurred at sites of monotonous or short tandem repeating sequences, and two of these sites, where there was a run of six Gs and four As, were hot spots. Three hot spot sites of base substitutions were found in HCT-15; two of them at splice acceptor sites, the other at the CpG site shared with HCT116. The distinct mutation spectra of the HCT116 and HCT-15 cell lines may reflect functional differences in the hMLH1 and GTBP gene products in mismatch repair. The gene product GTBP may be involved in the preferential repair of base mismatches, and MLH1 in the repair of both base mismatches and deletions/insertions of less than a few bases. These results suggest that mismatch repair deficiency affects the microsatellite stability as widely reported in colorectal tumour cells, but that it may not severely affect chromosome integrity as the karyotypes of these tumour cells are, unlike other tumour cells, relatively stable. PMID- 9214594 TI - Grafting assay distinguishes promotion sensitive from promotion resistant JB6 cells. AB - The JB6 mouse epidermal cell system has been used extensively as an in vitro transformation model for the study of tumor promotion. The standard JB6 cell assay for promotion of transformation is carried out in soft agar or other anchorage independent conditions. The present study was directed to the development of an in vivo model to distinguish the promotion resistant (P-) and promotion sensitive (P+) progression phenotypes. Results indicate that the grafting assay distinguishes P- and P+ cells in vivo with P+ but not P- cells forming tumors within 7-9 weeks. Expression of dominant negative mutant jun TAM67 blocks both anchorage independent transformation response and graft bed tumor formation by P+ cells, suggesting that the requirement for AP-1 activation in transformation now applies in vivo. Expression of mutated p53 produced a gain of P+ phenotype in P- cells in vitro, but not in vivo. Histochemical and Northern blot analysis for expression of various keratinocyte markers revealed no evidence for expression, suggesting a loss of keratinocyte markers following establishment in culture. In summary, the skin-grafting assay described in this study appears to be a valid in vivo assay for distinguishing the preneoplastic progression phenotypes represented by JB6 P- and P+ cells. PMID- 9214595 TI - Increased expression of the P27KIP1 protein in human esophageal cancer cell lines that over-express cyclin D1. AB - In the present study we have characterized eight human esophageal squamous carcinoma cell lines for levels of expression of cyclins D1, E, A and B1; CDKs 1, 2 and 4; the CDK inhibitors p16INK4, p21WAF1 and p27KIP1; the retinoblastoma (Rb) protein; and in vitro CDK2- and CDK4-associated kinase activity; and also compared the growth properties of these cell lines. The level of the cyclin D1 protein varied by over 30-fold amongst the eight cell lines. The high level in two cell lines was associated with amplification of this gene, but in three cell lines it was due to post-transcriptional events. Amongst the eight cell lines there was a significant correlation between the levels of cyclin D1, Rb and p27KIP1 proteins, and CDK4-associated kinase activity. Furthermore, when an exogenous cyclin D1 cDNA was over-expressed in the EC109 cell line by transfection, this led to increased expression of both Rb and p27KIP1. There was, however, no correlation between the level of cyclin D1 expression and the cell doubling times, duration of the G1 phase, or colony-forming efficiency in agar. Two of the cell lines displayed a high level of the cyclin E protein, low levels of cyclin D1, lacked expression of the Rb protein and expressed high levels of the p16INK4 protein. One of these cell lines displayed amplification of the latter gene. There was no correlation between the levels of cyclins E or A and in vitro CDK2 kinase activity, but CDK2 kinase activity was inversely correlated with the duration of the G1 phase of the cell cycle. Taken together, these studies indicate marked heterogeneity in the expression of cell cycle-related proteins amongst a series of esophageal carcinoma cell lines. The correlation between the levels of the cyclin D1, Rb and p27Kip1 proteins suggest the existence of a homeostatic feedback loop between positive and negative acting components of the cell cycle machinery. PMID- 9214596 TI - Altered gap junctional intercellular communication in neoplastic rat esophageal epithelial cells. AB - Gap junctional intercellular communication (GJIC) is reduced in many neoplastic cells, but few data exist for esophageal neoplasms. GJIC was examined by fluorescent dye microinjection in two nontumorigenic and two highly tumorigenic rat esophageal epithelial cell lines. All lines expressed high levels of dye coupling in homologous cell culture. In cocultures of nontumorigenic and tumorigenic cells, however, only one of six cell combinations displayed significant heterologous GJIC. Northern, Western, and immunohistochemical analyses indicated that all four cell lines expressed comparable levels of connexin43 (Cx43), but not connexin32 or connexin26, and formed Cx43-containing gap junction plaques at cell-cell interfaces. Immunostaining of rat esophageal frozen sections demonstrated that esophageal epithelial cells expressed Cx43 in vivo. In normal epithelium, the highest expression was seen in the basal cells and little suprabasal staining was evident. In preneoplastic and neoplastic lesions of the esophageal epithelium which were induced by treating rats with N nitrosomethylbenzylamine, Cx43 staining of the basal layer was also seen but appeared to be more diffuse compared to normal epithelium. In addition, suprabasal Cx43 staining was apparent in dysplastic and papillomatous lesions. These results indicate that Cx43 is expressed in normal and neoplastic rat esophageal cells and that the cells exhibit extensive homologous GJIC, but little heterologous GJIC. This lack of heterologous GJIC may be due to differences in cell adhesion proteins or other factors. PMID- 9214597 TI - Over expression of vascular endothelial growth factor and its receptor during the development of estrogen-induced rat pituitary tumors may mediate estrogen initiated tumor angiogenesis. AB - Estrogens, which have been associated with several types of human and animal cancers, can induce tumor angiogenesis in the pituitary of Fischer 344 rats. The mechanistic details of tumor angiogenesis induction, during estrogen carcinogenesis, are still unknown. To elucidate the role of estrogen in the regulation of tumor angiogenesis in the pituitary of female rats, the density of blood vessels was analysed using factor VIII related antigen (FVIIIRAg) immunohistochemistry and the expression of vascular endothelial growth factor/vascular permeability factor (VEGF/VPF) was examined by Western blot and immunohistochemical analysis. The expression of VEGF receptor (VEGFR-2/Flk-1/KDR) was also examined by immunohistochemistry. The results demonstrated that 17beta estradiol (E2) induces neovascularization, as well as the growth and enlargement of blood vessels after 7 days of exposure. The high tumor angiogenic potential was associated with an elevated VEGF/VPF protein expression in the E2 exposed pituitary of ovariectomized (OVEX) rats. VEGF/VPF and FVIIIRAg immunohistochemistry and endothelial specific lectin (UEA1) binding studies, indicate that the elevation of VEGF protein expression initially occurred in both blood vessels and non-endothelial cells. After 15 days of E2 exposure, VEGF/VPF protein expression, in the non-endothelial cell population, sharply declined and was restricted to the blood vessels. The function of non-endothelial-derived VEGF is not clear. Furthermore, immunohistochemical studies demonstrated that VEGFR-2 (flk-1/KDR), expression was elevated significantly in the endothelial cells of microblood vessels after 7 days of E2 exposure. These findings suggest that over expression of VEGF and its receptor (VEGFR-2) may play an important role in the initial step of the regulation of estrogen induced tumor angiogenesis in the rat pituitary. PMID- 9214598 TI - Ki-ras mutations are an early event and correlate with tumor stage in transplacentally-induced murine lung tumors. AB - A previous study from this laboratory demonstrated that treatment of pregnant mice with 3-methylcholanthrene (MC) caused lung tumors in the offspring at 1 year after birth, the incidence of which correlated with fetal inducibility of Cyp1a1. Analysis by PCR amplification and allele-specific hybridization (ASO) of paraffin embedded tumors generated from that study revealed the presence of point mutations in exon 1 of the Ki-ras gene. This work has now been expanded by PCR amplification and ASO analysis of 31 additional lesions. Point mutations were found in 37 of the 47 (79%) lesions analyzed in this and the previous study, the majority of which were G-->T transversions in the first or second base of codon 12. The mutational spectrum appeared to be dependent on the relative stage of differentiation of the lesion, as both the incidence of mutation and type of mutation produced correlated with malignant progression. Mutations occurred in 60% of the hyperplasias, 80% of the adenomas and 100% of the adenocarcinomas. In the lesions with mutations, GLY12-->CYS12 transversions occurred in 100% of the hyperplasias, 42% of the adenomas and 14% of the adenocarcinomas. The GLY12- >VAL12 transversions occurred in none of the hyperplasias, 42% of the adenomas and 57% of the adenocarcinomas. The remaining mutations, which consisted of ASP12 transitions and ARG13 transversions, occurred only in adenomas (17%) and adenocarcinomas (29%). Between this study and our previous analyses, the identity of the mutations obtained by ASO were confirmed by sequence analysis of eight of the 37 lesions that harbored mutations at the Ki-ras gene locus. There were no differences in the type or incidence of mutations relative to the metabolic phenotype or sex of the mice. These data suggest that mutational activation of the Ki-ras gene locus is an early event in transplacental lung tumorigenesis, and that the type of mutations produced by exposure to chemical carcinogens can influence the carcinogenic potential of the tumor. This may have prognostic significance in determining the malignant progression of the neoplasm. PMID- 9214599 TI - Antigenic phenotypes common to rat oval cells, primary hepatocellular carcinomas and developing bile ducts. AB - The shared expression of monoclonal antibody-defined antigens by oval cells and by bile ducts, neoplastic nodules and primary hepatocellular carcinomas (PHC) has provided support for the ability of oval cells to undergo differentiation along ductular or hepatocyte lineages and/or to progress to hepatocellular carcinoma. With the aim of obtaining additional insight into this process, we have combined serial section and double labeling immunofluorescence analysis to determine if phenotypes expressed in vitro by four rat oval cell lines and the H5D.61 hepatocellular carcinoma cell line and in situ by ethionine-induced primary hepatocellular carcinomas reproduce antigenic patterns occurring during normal liver development. Analysis using monoclonal antibodies specific for the oval cell antigens OV6 and OC.2 and hepatocyte markers HBD.1 and H.4 defined subpopulations in four oval cell lines and neoplastic hepatocytes in PHC and H5D.61 with OC.2-/OV6+ and OC.2+/OV6+ phenotypes. Cells with an OC2+/OV6- phenotype were rarely observed in cell lines or primary tumors. In contrast, areas composed of OV6+/H.4+ cells were frequently found in PHC. Examination of fetal and neonatal rat livers demonstrated the stage-specific appearance of three of these phenotypes during liver development. The OC.2+/OV6- phenotype appeared transiently prior to embryonic day (ED) 18 in a subpopulation of HBD.1+ hepatoblasts. OV6 expression was first detected at ED18 on developing bile ducts that were negative for OC.2. These newly formed ducts rapidly acquired OC.2, starting with ducts in the hilar region and spreading outward towards the periphery. This OC.2 expression gradient persisted in the newborn rat liver but became more skewed towards doubly positive cells, with OC.2-/OV6+ cells being found primarily in the periphery. Hepatocytes expressing both OV6 and H.4 were not observed in fetal liver but appeared in neonatal liver in close proximity to OV6+ interlobular ducts. From these findings, it was concluded that oval cells and PHC display phenotypes representing normal stages in liver development, suggesting that oval cells and cells within ethionine-induced PHC are capable of initiating but are unable to complete pathways of hepatocytic or biliary differentiation. PMID- 9214600 TI - Carcinogenic polycyclic aromatic hydrocarbons increase intracellular Ca2+ and cell proliferation in primary human mammary epithelial cells. AB - Previous studies have shown that polycyclic aromatic hydrocarbons (PAHs) mobilize intracellular Ca2+ in human T cells by inositol trisphosphate-dependent mechanisms resulting from activation of phospholipase C-gamma by SRC-related protein tyrosine kinases, thereby mimicking antigen-receptor activation. Ca2+ appears to play an important second messenger role in growth factor control of cell proliferation in human mammary epithelial cells (HMEC), such as the epidermal growth factor receptor pathway. The purpose of the present studies was to determine if PAHs are able to increase intracellular Ca2+ in primary cultures of HMEC and increase cell proliferation. Two carcinogenic and two non carcinogenic PAHs were tested for their ability to increase intracellular Ca2+ in HMEC. The carcinogenic PAHs dimethylbenz[a]anthracene (DMBA) and benzo[a] pyrene (BaP) were able to cause Ca2+ elevation in HMEC at early time points (2 h) and caused sustained alterations in Ca2+ homeostasis (18 h). DMBA showed maximal effects at early time points (2 h), while BaP showed maximal effects on sustained Ca2+ (18 h). 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD), a potent dioxin and tumor promoter, produced maximal Ca2+ elevation at 2 h, with a return to near baseline levels by 6 h. The non-carcinogenic PAHs benzo[e]pyrene and anthracene did not significantly alter intracellular Ca2+ at any time point. alpha Naphthoflavone significantly reduced the Ca2+ response induced by BaP treatment, but not by DMBA or TCDD, suggesting that P450 1A or 1B metabolism of BaP may be important in the sustained Ca2+ elevating response. In evaluating the effects of BaP on HMEC proliferation, BaP was found to increase the number of cells recovered after 4 days in culture in the absence or presence of various concentrations of epidermal growth factor. These studies provide initial evidence that Ca2+ signaling may be associated with mitogenesis in HMEC, which may play a role in tumor promotion and progression produced by PAHs. PMID- 9214601 TI - Energy availability and mammary carcinogenesis: effects of calorie restriction and exercise. AB - The purpose of this experiment was to compare the carcinogenic response in the mammary gland among groups of rats whose energy metabolism had been modulated by restricting dietary calories and/or by increasing energy expenditure via exercise. Female F344 rats (n = 132) were injected i.p. with 1-methyl-1 nitrosomethylurea (50 mg/kg at 50 and 57 days of age) and were randomized into one of four treatment groups: (i) unrestricted, sedentary; (ii) calorie restricted, sedentary; (iii) unrestricted, exercised; (iv) calorie-restricted, exercised. The targeted level of calorie-restricted was 20% and exercise was achieved by treadmill-running (20 m/min at a 15% grade for 30 min, 5 days/week). During the 20.5 week study, rats were palpated twice a week for detection of mammary tumors and urine was collected for determination of 24-h cortical steroid excretion. At the end of the study, all mammary lesions were histologically classified. Carcass composition and carcass energy were determined. Mammary carcinogenesis was inhibited among calorie-restricted, sedentary rats compared with unrestricted, sedentary rats (79% inhibition, P < 0.001). No inhibition of carcinogenesis was observed among exercised rats (unrestricted or calorie restricted) relative to the unrestricted, sedentary rats. Within the present experimental design, exercise had no effect on carcinogenesis despite significant reductions of carcass fat and carcass energy among both groups of rats that exercised. Cortical steroid level was significantly higher only in calorie restricted, sedentary rats (P < 0.05). These results do not support the hypothesis that reductions of body weight gain, carcass fat or carcass energy are sufficient conditions to inhibit mammary carcinogenesis. The results do suggest that changes in urinary cortical steroid excretion may predict whether an energy related intervention is likely to alter mammary carcinogenesis. PMID- 9214602 TI - Plasma antioxidant vitamins, chronic hepatitis B virus infection and urinary aflatoxin B1-DNA adducts in healthy males. AB - Epidemiological evidence indicates that aflatoxin B1 (AFB1) intake is associated with an increased risk of hepatocellular carcinoma (HCC). The hepatocarcinogenesis is initiated by covalent binding of AFB1 to cellular DNA. To determine whether nutritional factors and hormonal status may influence the binding of AFB1 to hepatic DNA, a cross-sectional study was performed on a total of 42 male asymptomatic hepatitis B surface antigen (HBsAg) carriers and 43 male non-carriers in a cohort study on the multistage development of HCC in Taiwan. The major AFB1-DNA adduct in vivo, AFB1-N7-guanine, was measured by high performance liquid chromatography in urine. Urinary AFB1-N7-guanine was detectable in 40% of the subjects. HBsAg carriers had a higher detection rate of urinary AFB1-DNA adducts than non-carriers and the difference was statistically significant after multivariate adjustment. After taking into account the total AFB1 urinary metabolite level, chronic HBsAg carrier status, and other potential confounders, plasma levels of cholesterol, alpha-tocopherol, and alpha- and beta carotene were positively associated with the detection rate of the AFB1-DNA adducts in a dose-dependent manner, whereas plasma lycopene level was inversely related to the presence of the adducts in urine. The association of urinary AFB1 DNA adducts with the plasma levels of cholesterol, alpha-tocopherol, lycopene, and alpha- and beta-carotene was observed at both low and high exposure levels of AFB1. There was a synergistic interaction of plasma alpha-tocopherol with alpha- and beta-carotene on the adduct levels. No association with the adducts was found for plasma levels of retinol and testosterone. This study demonstrated different associations of antioxidant vitamins with AFB1-DNA adduct formation. The data consistent with our previous finding in cultured woodchuck hepatocytes that alpha tocopherol and beta-carotene enhanced AFB1-DNA adduct formation suggest that prospective investigation of the relationship between plasma micronutrients and risk of AFB1-related HCC is warranted. PMID- 9214603 TI - Involvement of polyamines in selenomethionine induced apoptosis and mitotic alterations in human tumor cells. AB - The efficacy of dietary selenium supplementation is currently being evaluated in intervention trials. However, the biological mechanisms underlying the cancer chemopreventive effects of selenium supplementation have yet to be elucidated. Selenium metabolism and polyamine biosynthesis are linked in their common requirement for S-adenosylmethionine. Selenomethionine was the predominant form of selenium in the dietary supplement, therefore we evaluated the anti tumorigenic effects of selenomethionine. We found that selenomethionine inhibited tumor growth (both in A549 lung and HT29 colon cancer cells) in a dose-dependent manner. At 24 and 72 h, polyamine content of A549 and HT29 cancer cell lines was decreased at doses that inhibited 50% of normal growth. Selenomethionine treatment induced apoptosis in both cancer cell lines. Exogenous spermine administration, which replenishes intracellular polyamine levels, prevented selenomethionine induced apoptosis. Selenomethionine administration to the cancer cell lines increased the number of cells in metaphase. This cell cycle effect appeared to be reversed with the co-administration of selenomethionine and spermine. These data suggested that at least part of the anti-carcinogenic effects of selenium supplementation might be due to a depletion in polyamine levels. This depletion of polyamines leads to an induction in apoptosis and perturbations in the cell cycle. PMID- 9214604 TI - Genetic polymorphism of CYP2D6 and lung cancer risk in African-Americans and Caucasians in Los Angeles County. AB - The well described genetic polymorphism of the CYP2D6 gene influences response to a wide variety of therapeutic agents metabolized by the CYP2D6 enzyme product. CYP2D6 also appears to play a role, along with other cytochrome P450 enzymes, in the metabolic activation of the tobacco specific nitrosamine, NNK, as well as metabolism of nicotine to cotinine. While impaired activity of CYP2D6 was strongly protective against lung cancer in some studies, primarily based on phenotyping, the literature is conflicting. The molecular basis of CYP2D6 deficiency is now well understood, enabling the use of genotyping to classify individuals. We therefore examined whether lung cancer risk is reduced by the presence of four CYP2D6 alleles associated with impaired activity due to an inactivating mutation--CYP2D6*4, CYP2D6*3, CYP2D6*5 and CYP2D6*16--among 341 incident cases of lung cancer and 710 population controls of Caucasian or African American ethnicity in Los Angeles County, California. We did not confirm a strong association between the presence of these inactivating alleles and lung cancer risk [odds ratio (OR) = 0.90, 95% confidence interval (CI) 0.60-1.35 for Caucasians], although there was a small decreased risk among the African Americans (OR = 0.66, 95% CI 0.38-1.14). Among smokers, when the data are stratified according to lifetime smoking history, there is a suggestion of an association limited to Caucasian smokers of <35 pack-years, the median for all smokers in these data (OR = 0.49, 95% CI 0.23-1.04). However, among African American smokers, who smoke less than Caucasians, the association did not differ between smoking categories. We also examined the possible role of additional copies of the CYP2D6 gene, which lead to enhanced CYP2D6 activity, in increasing lung cancer risk. Among controls the prevalence of having more than two copies of the CYP2D6 gene and no inactivating alleles was 4.3% for Caucasians and 4.9% for African-Americans. Relative to subjects with an inactivating allele, those with an additional copy of the CYP2D6 gene and no inactivating alleles may be at increased risk of lung cancer, particularly for adenocarcinoma (OR = 3.61, 95% CI 1.08-11.7 for African-Americans and OR = 2.20, 95% CI 0.69-6.0 for Caucasians). Our data suggest that the CYP2D6 genetic polymorphism is not the strong risk factor for lung cancer suggested by some studies of phenotype, but may play a minor role. PMID- 9214605 TI - Androgen responsive adult human prostatic epithelial cell lines immortalized by human papillomavirus 18. AB - Prostate cancer and benign tumors of the prostate are the two most common neoplastic diseases in men in the United States, however, research on their causes and treatment has been slow because of the difficulty in obtaining fresh samples of human tissue and a lack of well characterized cell lines which exhibit growth and differentiation characteristics of normal prostatic epithelium. Non neoplastic adult human prostatic epithelial cells from a white male donor were immortalized with human papillomavirus 18 which resulted in the establishment of the RWPE-1 cell line. Cells from the RWPE-1 cell line were further transformed by v-Ki-ras to establish the RWPE-2 cell line. The objectives of this study were to: (1) establish the prostatic epithelial origin and androgen responsiveness of RWPE 1 and RWPE-2 cell lines; (2) examine their response to growth factors; and (3) establish the malignant characteristics of the RWPE-2 cell line. Immunoperoxidase staining showed that both RWPE-1 and RWPE-2 cells express cytokeratins 8 and 18, which are characteristic of luminal prostatic epithelial cells, but they also coexpress basal cell cytokeratins. These cell lines show growth stimulation and prostate specific antigen (PSA) and androgen receptor (AR) expression in response to the synthetic androgen mibolerone, which establishes their prostatic epithelial origin. Both cell lines also show a dose-dependent growth stimulation by EGF and bFGF and growth inhibition when exposed to TGF-beta, however, the transformed RWPE-2 cells are less responsive. RWPE-1 cells neither grow in agar nor form tumors when injected into nude mice with or without Matrigel. However, RWPE-2 cells form colonies in agar and tumors in nude mice. In the in vitro invasion assay, RWPE-1 cells are not invasive whereas RWPE-2 cells are invasive. Nuclear expression of p53 and Rb proteins was heterogeneous but detectable by immunostaining in both cell lines. The RWPE-1 cells, which show many normal cell characteristics, and the malignant RWPE-2 cells, provide useful cell culture models for studies on prostate growth regulation and carcinogenesis. PMID- 9214606 TI - Acinar differentiation by non-malignant immortalized human prostatic epithelial cells and its loss by malignant cells. AB - Invasive prostatic carcinomas and prostatic intraepithelial neoplasia (PIN) are characterized by a loss of normal cell organization, cell polarity, and cell:cell and cell:basement membrane adhesion. The objective of this study was to establish in vitro three-dimensional (3-D) cell models which can be used to investigate mechanisms involved in acinar morphogenesis and differentiation in normal prostatic epithelium and their abnormalities in cancer cells. The process of acinar morphogenesis, including structural and functional differentiation, was investigated by culture on basement membrane gels (Matrigel). The human papillomavirus 18 immortalized, non-tumorigenic cell line RWPE-1, the v-Ki-ras transformed, tumorigenic RWPE-2 cell line derived from RWPE-1 cells (see previous paper pp. 1221-1229) and the human prostatic carcinoma cell line DU-145 were used. When cultured on Matrigel, RWPE-2 cells remain as single cells or form small aggregates and DU-145 cells form large amorphous cell aggregates without any organization or lumen. In contrast, RWPE-1 cells form acini of polarized epithelium with a distinct lumen, show a distinct laminin basement membrane, and express alpha6beta1 integrins at their basal end. Exposure to conditioned medium from NIH 3T3 cultures accelerates glandular morphogenesis. Parallel cultures maintained as monolayers on plastic remain as monolayers. In the presence of the synthetic androgen mibolerone, acinar cells express prostate specific antigen (PSA) as determined by immunostaining. We conclude that normal prostate cells can undergo acinar morphogenesis while tumorigenic cells have lost this ability. The 3-D cultures provide physiologically relevant in vitro models for elucidating regulation of growth, morphogenesis and differentiation in the normal human prostate, for defining heterotypic interactions in benign prostatic hyperplasia and for establishing the basis for the loss of normal cell organization in early neoplastic lesions such as PIN as well as during tumor progression in prostate cancer. PMID- 9214607 TI - Reductive metabolism of 4-nitrobiphenyl by rat liver fraction. AB - The metabolites of 4-nitrobiphenyl (4-NBP) were studied using S-9 under anaerobic conditions. Ten metabolites were isolated and tentatively identified by high performance liquid chromatography/atmospheric pressure chemical ionization/mass spectrometry (HPLC/APCI/MS), which is a novel efficient technqiue for analyses of metabolites. HPLC retention times and UV spectra have been also used for the confirmation. Among them, 4-aminobiphenyl (4-ABP, 79% of total metabolites) and hydroxylaminobiphenyls were found to be the major metabolites, and 4 acetylaminobiphenyl (4-AABP), N-hydroxy-4-acetylaminobiphenyl (4-AABP-N-OH), x-OH 4-nitrobiphenyl (4-NBP-x-OH), biphenylene and N-formyl-4-aminobiphenyl (N-formyl 4-ABP) were the minor metabolites. Based on the metabolities identified, different metabolic pathways of 4-NBP in rat liver, including N-hydroxylation, N acetylation, N-formylation, de-nitro-group biotransformation and ring hydroxylation were proposed to elucidate the mechanisms of toxification and detoxification. In addition, experimental conditions such as incubation time and S-9 amounts were optimized. PMID- 9214608 TI - p53 protein expression and increased SSCP mobility shifts in the p53 gene in normal urothelium cultured from smokers. AB - This study provides evidence of a significantly (P = 0.018) increased level of expression of the stable conformation of p53 in normal urothelial cells, cultured in vitro from bladder biopsies obtained from normal smokers without malignant disease of any site. With two significant exceptions, non-smokers showed low or no expression of this protein. Past smokers appeared to segregate into high or low p53 expressers, but the expression was not correlated with years since quitting smoking or with pack years smoked. The mean data in this group were not quite significantly different (P = 0.08) from the non-smoker group, due to the wide inter-patient variation. For most of the smoker group, pack years correlated with p53 expression with a mean unit of 1.7 +/- 0.37% p53 per pack year but there was a small group of very heavy smokers who showed lower than expected expression (approximately 0.3-0.8% p53 per pack year). These were statistical outliers (Grubbs test). No explanation could be found for this. Over-expression of p53 protein, often correlates with mutations in the gene, but may also indicate that breakdown of wild-type p53 has slowed. SSCP analysis of the biopsy material was not possible on all patients due to ethical constraints on the amounts of tissue which could be taken but in the cases where it was possible the association between loss of p53 protein function and mobility shifts in p53 exons 5-8 was confirmed with smokers having 3.5 times the number of mobility shifts detected in non-smoker DNA. Thus the results may point to a role for the early abrogation of p53 protein function in bladder carcinogenesis induced by cigarette smoking. PMID- 9214609 TI - Estrogen-nucleic acid adducts: guanine is major site for interaction between 3,4 estrone quinone and COIII gene. AB - The carcinogenicity of estrogens in rodents and man has been attributed to either alkylation of cellular macromolecules and/or redox-cycling, generation of active radicals and DNA damage. Metabolic activation of estradiol leading to the formation of catechol estrogens is believed to be a prerequisite for its genotoxic effects. 4-Hydroxyestradiol is a potent inducer of tumors in hamsters. Previous studies have shown that 3,4-estrone quinone (3,4-EQ) can redox-cycle and is capable of inducing exclusively single strand DNA breaks in MCF-7 breast cancer cells, as well as react with various nucleophiles including amino acids and nucleic acids to give Michael addition products. In this paper we examined the nature of the interaction of 3,4-EQ with COIII gene and analysed the estrogen DNA adducts by 32P-post-labeling. The reaction of 3,4-EQ with the COIII gene followed by polymerase arrest assay showed several stop sites in which guanine was preferentially attacked by 3,4-EQ and, to a lesser extent, with Ade, Cyt and Thy. 32P-Post-labeling analysis of the reaction of 3,4-EQ with COIII gene gave one major adduct which was found to be identical to that obtained from reaction of dGMP with 3,4-EQ. The observation that obstruction of in vitro replication of COIII template bound to 3,4-EQ suggests that estrogen quinone adducted lesions can arrest DNA polymerase. These results indicate that 3,4-EQ may be genotoxic and may provide one possible explanation for the carcinogenic effects of estrogens. PMID- 9214610 TI - Cytogenetic and molecular genetic analysis of tumorigenic human bronchial epithelial cells induced by radon alpha particles. AB - To establish a cell culture model for lung carcinogenesis, independent populations of the human papillomavirus 18-immortalized human bronchial epithelial cell line BEP2D were treated with high linear energy transfer radon simulated alpha-particles, expanded and xenotransplanted into Nu/Nu mice. Six independent cell lines were established from tumors that developed from three separate radiation treatments as follows: treatment (Tx) 1 (30 cGy--two doses), H2BT, Tx 2 (30 cGy--single dose), R30T1L, R30T2 and R30T3L, Tx 3 (30 cGy--single dose), H1ATN and H1ATBA1. Cytogenetic analysis revealed common changes in all tumor lines: loss of the Y chromosome (ch), one of three copies of ch8, one of three copies of ch14, and one of two copies of ch4p16-pter and ch11p15-pter. Analysis of polymerase chain reaction-amplified short tandem repeats of informative loci confirmed the loss of chY in all lines and loss of heterozygosity (LOH) at eight loci spanning the length of ch8 in all lines from Tx's 1 and 2. Our data support previous studies indicating the presence of tumor suppressor genes on ch8. LOH also was confirmed on ch14 at locus D14S306 in all cell lines from Tx 2 and in one of two lines from Tx 3. This region, 14q12-q13, may contain changes in one of the five known somatostatin receptor genes (SSTR1). No LOH was detected at any of the informative loci tested for on ch4 or ch11. PMID- 9214611 TI - Enhancement of cell proliferation and prostaglandin biosynthesis by 1,8 dihydroxyanthraquinone in the rat large intestine. AB - The effects of 1,8-dihydroxyanthraquinone (DHAQ), a stimulant laxative named danthron, on cell kinetics and prostaglandin (PG) biosynthesis in the gastrointestinal tract were investigated in male 8-week-old F344 rats divided into three groups, each consisting of 10 animals. The animals in groups one, two and three were respectively given diets supplemented with 0%, 0.1% and 0.2% DHAQ for 24 days. PGE2 levels in the colorectal mucosa were significantly (P < 0.05 and 0.001) elevated after DHAQ treatment and showed some evidence of a dependence of DHAQ dose, consistent with the plasma PGE2 levels. BrdU-labeling indices in the large intestinal epithelium were also significantly (P < 0.01) increased, although the other portions of the gut such as the stomach and small intestine were not significantly affected. Excretion of the main urinary metabolite of PGE (PGE-MUM) was significantly (P < 0.001 or 0.01) increased whereas the urinary PGE2 concentration and total PGE2 excretion were not changed. Thus the results of the present study clearly indicate enhancement of cell proliferation by DHAQ in the large intestine epithelia, correlated with increased PGE2 levels in the large intestinal mucosa as well as the plasma, and possible support for the conclusion that quantitative analysis of urinary PGE-MUM, but not PGE2 itself, offer a useful approach for biomonitoring exposure to such stimulant laxatives. PMID- 9214612 TI - Butyrate can act as a stimulator of growth or inducer of apoptosis in human colonic epithelial cell lines depending on the presence of alternative energy sources. AB - In vivo, butyrate is a major energy source for the colonic epithelium and is thought to stimulate proliferation. In contrast, butyrate in vitro has been shown to inhibit proliferation and induce differentiation and apoptosis in colonic epithelial cells. Most colon cell cultures are grown in medium containing high concentrations of glucose, whereas in vivo, the main energy source used by the colon cells is butyrate. The aim of this study was to determine whether the apparent contrasting roles of butyrate in vivo and in vitro could be as a consequence of differences in glucose availability. The sensitivity of two human colorectal tumour cell lines, one adenoma (S/RG/C2) and one carcinoma (HT29) to butyrate-induced growth inhibition and apoptosis was investigated to determine whether these cellular effects were altered under glucose depleted culture conditions. Glucose depletion resulted in increased apoptosis in both cell lines in the absence of butyrate. Butyrate in standard culture conditions (containing 25 mM glucose and 1 mM pyruvate) inhibited growth and induced apoptosis in both cell lines. However, low concentrations of butyrate in glucose depleted culture conditions (i.e. standard growth medium without glucose and pyruvate supplements) were found to reduce apoptosis induced by glucose deprivation and increase cell yield in both cell lines. The results show that in glucose depleted culture conditions, butyrate at low concentrations (0.5 mM for S/RG/C2, and 0.5 and 2 mM for HT29 cells) was found to be growth stimulatory whereas in the presence of glucose, these same concentrations of butyrate induced apoptosis. Thus, whether butyrate is growth stimulatory or growth inhibitory may depend on the availability of other energy sources. These observations may, in part, provide an explanation for the apparent opposite effects of butyrate on proliferation reported in vivo and in vitro. PMID- 9214613 TI - Serum, plasma and paraffin-embedded tissues as sources of DNA for studying cancer susceptibility genes. AB - The ability to isolate DNA from archived human serum, plasma and paraffin embedded human tissues enhances opportunities to study breast, lung and other cancer risk factors. We report herein a simple and fast protocol for the extraction of genomic DNA from these sources. Using a phenol-based extraction method, the recovery for DNA is quantitative and reproducible. DNA yields in serum (250 microl) were between 162 and 1060 ng (n = 18 subjects), in plasma (250 microl) were between 165 and 375 ng (n = 5 subjects) and in embedded tissues (5 microm thick sections for ethanol fixed, and between 5- and 20-microm sections for formaldehyde fixation) were between 1 microg and 11.7 microg (n = 32 subjects). The extraction method was combined with newly designed PCR-based assays for cancer susceptibility marker genes such as CYP1A1 (exon 7), CYP2E1 (Dra1, Rsa1), GSTM1 and NAT2 [NAT2*5A (C481T), NAT2*6A (G590A), NAT2*7A (G857A)]. Genotyping results from the serum and paraffin-embedded tissues compared favorably to results from archived freshly frozen tissues, where concordance was 98% for serum, 100% for ethanol-fixed embedded tissues, and 97% for formaldehyde fixed and paraffin-embedded tissues. This facile method will allow for the use of archived tissue samples of prospective cohort and other studies where intact DNA was not previously available. PMID- 9214614 TI - Chemical carcinogens and antigens contribute to cutaneous tumor promotion by depleting epidermal Langerhans cells. AB - Epidermal Langerhans cells (LC) are an integral component of the skin immune system as they initiate immune responses to a variety of antigens, including tumor antigens. When skin is exposed to carcinogenic doses of ultraviolet-B irradiation, chemical carcinogens or tumor promoters there is a significant reduction of LC density. This causes the skin to be immunocompromised and provides an opportunity for aberrant cells to escape immune detection and develop into tumors. Consequently LC depletion is a key event associated with the pathogenesis of skin cancer. We propose that LC depletion contributes to tumor promotion and therefore any agents that reduce LC number, e.g. the contact sensitizing antigen 2,4,6-trinitrochlorobenzene (TNCB), may also contribute to tumor promotion. This proposal was evaluated in cutaneous carcinogenesis by treating mouse skin with a tumor initiating dose of the carcinogen 7,12 dimethylbenz[a]anthracene (DMBA) followed by a tumor promoter. The initiating dose of DMBA did not cause LC depletion or tumor development. However, if the DMBA-treated skin was then exposed to a concentration of TNCB that caused LC depletion, skin tumors developed. This is analogous to the classical initiator/promoter system with an LC-depleting dose of TNCB contributing to tumor promotion. Further, this promotion effect was independent of the commencement time of the promoter application, as 2% TNCB applied either 1 or 12 weeks after DMBA initiation induced tumor development. Analysis of the association of LC depletion with immunosuppression and tumor promotion, showed that these events were linked, irrespective of the agent that caused the depletion. It is therefore concluded that LC depletion and local immunosuppression are important aspects of tumor promotion in cutaneous carcinogenesis and non-carcinogenic agents may have tumor promoter activities. PMID- 9214615 TI - The thrombin E192Q-BPTI complex reveals gross structural rearrangements: implications for the interaction with antithrombin and thrombomodulin. AB - Previous crystal structures of thrombin indicate that the 60-insertion loop is a rigid moiety that partially occludes the active site, suggesting that this structural feature plays a decisive role in restricting thrombin's specificity. This restricted specificity is typified by the experimental observation that thrombin is not inhibited by micromolar concentrations of basic pancreatic trypsin inhibitor (BPTI). Surprisingly, a single atom mutation in thrombin (E192Q) results in a 10(-8) M affinity for BPTI. The crystal structure of human thrombin mutant E192Q has been solved in complex with BPTI at 2.3 A resolution. Binding of the Kunitz inhibitor is accompanied by gross structural rearrangements in thrombin. In particular, thrombin's 60-loop is found in a significantly different conformation. Concomitant reorganization of other surface loops that surround the active site, i.e. the 37-loop, the 148-loop and the 99-loop, is observed. Thrombin can therefore undergo major structural reorganization upon strong ligand binding. Implications for the interaction of thrombin with antithrombin and thrombomodulin are discussed. PMID- 9214616 TI - Targeted expression of MYCN causes neuroblastoma in transgenic mice. AB - The proto-oncogene MYCN is often amplified in human neuroblastomas. The assumption that the amplification contributes to tumorigenesis has never been tested directly. We have created transgenic mice that overexpress MYCN in neuroectodermal cells and develop neuroblastoma. Analysis of tumors by comparative genomic hybridization revealed gains and losses of at least seven chromosomal regions, all of which are syntenic with comparable abnormalities detected in human neuroblastomas. In addition, we have shown that increases in MYCN dosage or deficiencies in either of the tumor suppressor genes NF1 or RB1 can augment tumorigenesis by the transgene. Our results provide direct evidence that MYCN can contribute to the genesis of neuroblastoma, suggest that the genetic events involved in the genesis of neuroblastoma can be tumorigenic in more than one chronological sequence, and offer a model for further study of the pathogenesis and therapy of neuroblastoma. PMID- 9214617 TI - The extent of affinity maturation differs between the memory and antibody-forming cell compartments in the primary immune response. AB - Immunization with protein-containing antigens results in two types of antigen specific B cell: antibody forming cells (AFCs) producing antibody of progressively higher affinity and memory lymphocytes capable of producing high affinity antibody upon re-exposure to antigen. The issue of the inter relationship between affinity maturation of memory B cells and AFCs was addressed through analysis of single, antigen-specific B cells from the memory and AFC compartments during the primary response to a model antigen. Only 65% of splenic memory B cells were found capable of producing high affinity antibody, meaning that low affinity cells persist into this compartment. In contrast, by 28 days after immunization all AFCs produced high affinity antibody. We identified a unique, persistent sub-population of bone marrow AFCs containing few somatic mutations, suggesting they arose early in the response, yet highly enriched for an identical affinity-enhancing amino acid exchange, suggesting strong selection. Our results imply that affinity maturation of a primary immune response occurs by the early selective differentiation of high affinity variants into AFCs which subsequently persist in the bone marrow. In contrast, the memory B-cell population contains few, if any, cells from the early response and is less stringently selected. PMID- 9214618 TI - Specific interaction between OutD, an Erwinia chrysanthemi outer membrane protein of the general secretory pathway, and secreted proteins. AB - OutD is an outer membrane component of the main terminal branch of the general secretory pathway (GSP) in Erwinia chrysanthemi. We analyzed the interactions of OutD with other components of the GSP (Out proteins) and with secreted proteins (PelB, EGZ and PemA). OutD is stabilized by its interaction with another GSP component, OutS. The 62 C-terminal amino acids of OutD are necessary for this interaction. In vivo formation of OutD multimers, up to tetramers, was proved after the dissociation in mild conditions of the OutD aggregates formed in the outer membrane. Thus, OutD could form a channel-like structure in the outer membrane. We showed that OutD is stabilized in vivo when co-expressed with Out secreted proteins. This stabilization results from the formation of complexes that were detected in experiments of co-immunoprecipitation and co-sedimentation in sucrose density gradients. The presence of the N-terminal part of OutD is required for this interaction. The interaction between OutD and the secreted protein PelB was confirmed in vitro, suggesting that no other component of the GSP is required for this recognition. No interaction was observed between the E. carotovora PelC and the E. chrysanthemi OutD. Thus, the interaction between GspD and the secreted proteins present in the periplasm could be the key to the specificity of the secretion machinery and a trigger for that process. PMID- 9214619 TI - A novel SNARE complex implicated in vesicle fusion with the endoplasmic reticulum. AB - Intracellular vesicular traffic is controlled in part by v- and t-SNAREs, integral membrane proteins which allow specific interaction and fusion between vesicles (v-SNAREs) and their target membranes (t-SNAREs). In yeast, retrograde transport from the Golgi complex to the ER is mediated by the ER t-SNARE Ufe1p, and also requires two other ER proteins, Sec20p and Tip20p, which bind each other. Although Sec20p is not a typical SNARE, we show that both it and Tip20p can be co-precipitated with Ufe1p, and that a growth-inhibiting mutation in Ufe1p can be compensated by a mutation in Sec20p. Furthermore, Sec22p, a v-SNARE implicated in forward transport from ER to Golgi, co-precipitates with Ufe1p and Sec20p, and SEC22 acts as an allele-specific multicopy suppressor of a temperature-sensitive ufe1 mutation. These results define a new functional SNARE complex, with features distinct from the plasma membrane and cis-Golgi complexes previously identified. They also show that a single v-SNARE can be involved in both anterograde and retrograde transport, which suggests that the mere presence of a particular v-SNARE may not be sufficient to determine the preferred target for a transport vesicle. PMID- 9214620 TI - Kinesin is essential for cell morphogenesis and polarized secretion in Neurospora crassa. AB - Kinesin is a force-generating molecule that is thought to translocate organelles along microtubules, but its precise cellular function is still unclear. To determine the role of kinesin in vivo, we have generated a kinesin-deficient strain in the simple cell system Neurospora crassa. Null cells exhibit severe alterations in cell morphogenesis, notably hyphal extension, morphology and branching. Surprisingly, the movement of organelles visualized by video microscopy is hardly affected, but apical hyphae fail to establish a Spitzenkorper, an assemblage of secretory vesicles intimately linked to cell elongation and morphogenesis in Neurospora and other filamentous fungi. As cell morphogenesis depends on polarized secretion, our findings demonstrate that a step in the secretory pathway leading to cell shape determination and cell elongation cannot tolerate a loss of kinesin function. The defect is suggested to affect the transport of small, secretory vesicles to the site involved in protrusive activity, resulting in the uncoordinated insertion of new cell wall material over much of the cell surface. These observations have implications for the presumptive function of kinesin in more complex cell systems. PMID- 9214621 TI - Dimer model for the microfibrillar protein fibulin-2 and identification of the connecting disulfide bridge. AB - Fibulin-2 is a novel extracellular matrix protein frequently found in close association with microfibrils containing either fibronectin or fibrillin. The entire protein and its predicted domains were obtained as recombinant products and examined by ultracentrifugation and electron microscopy. This demonstrated a disulfide-linked homodimer of 175 kDa subunits. Partial reduction to monomers identified specifically an odd Cys574 residue responsible for dimer formation in one of three anaphylatoxin-like modules that constitute the central globular domain I (13 kDa) of fibulin-2. Furthermore, a Cys574-Ser mutation abolished disulfide connection but not non-covalent dimerization of fibulin-2. The C terminal region (85 kDa) was shown to represent a 35-nm-long rod consisting of 11 calcium-binding EGF-like modules (domain II) and a small terminal globe (domain III). The unique N-terminal domain N (55 kDa) was also rod-shaped (approximately 38 nm) and rich in galactosamine indicating extensive O-glycosylation. A dimer model is proposed indicating mainly a rod-like shape of 80 nm length based on an anti-parallel association of two subunits through their domains I. This model also implies alignment of domains II and N between different subunits. This was demonstrated by surface plasmon resonance assay which showed a distinct interaction between domains N and II with a Kd of approximately 0.7 microM. PMID- 9214622 TI - p140mDia, a mammalian homolog of Drosophila diaphanous, is a target protein for Rho small GTPase and is a ligand for profilin. AB - Rho small GTPase regulates cell morphology, adhesion and cytokinesis through the actin cytoskeleton. We have identified a protein, p140mDia, as a downstream effector of Rho. It is a mammalian homolog of Drosophila diaphanous, a protein required for cytokinesis, and belongs to a family of formin-related proteins containing repetitive polyproline stretches. p140mDia binds selectively to the GTP-bound form of Rho and also binds to profilin. p140mDia, profilin and RhoA are co-localized in the spreading lamellae of cultured fibroblasts. They are also co localized in membrane ruffles of phorbol ester-stimulated sMDCK2 cells, which extend these structures in a Rho-dependent manner. The three proteins are recruited around phagocytic cups induced by fibronectin-coated beads. Their recruitment is not induced after Rho is inactivated by microinjection of botulinum C3 exoenzyme. Overexpression of p140mDia in COS-7 cells induced homogeneous actin filament formation. These results suggest that Rho regulates actin polymerization by targeting profilin via p140mDia beneath the specific plasma membranes. PMID- 9214623 TI - Co-amplification of the gamma-glutamylcysteine synthetase gene gsh1 and of the ABC transporter gene pgpA in arsenite-resistant Leishmania tarentolae. AB - Resistance to the oxyanion arsenite in the parasite Leishmania is multifactorial. We have described previously the frequent amplification of the ABC transporter gene pgpA, the presence of a non-PgpA thiol-metal efflux pump and increased levels of glutathione and trypanothione in resistant cells. Other loci are also amplified, although their role in resistance is unknown. By gene transfection, we have characterized one of these novel genes. It corresponds to gsh1, which encodes gamma-glutamylcysteine synthetase, an enzyme involved in the rate limiting step of glutathione biosynthesis. Transfection of gsh1 in wild-type cells increased the levels of glutathione and trypanothione to levels found in resistant mutants. These transfectants were not resistant to metals. However, when gsh1 was transfected in partial revertants, it conferred resistance. As pgpA is frequently co-amplified with gsh1, we co-transfected the two genes into both wild-type and partial revertants. Arsenite resistance levels in wild-type cells could be accounted for by the contribution of PgpA alone. In the partial revertant, the gsh1 and pgpA gene product acted synergistically. These results support our previous suggestion that PgpA recognizes metals conjugated to thiols. Furthermore, amplification of gsh1 overcomes the rate-limiting step in the synthesis of trypanothione, contributing to resistance. In addition, the results suggest that at least one more factor acts synergistically with the gsh1 gene product. PMID- 9214624 TI - Subunit interactions in ABC transporters: a conserved sequence in hydrophobic membrane proteins of periplasmic permeases defines an important site of interaction with the ATPase subunits. AB - The cytoplasmic membrane proteins of bacterial binding protein-dependent transporters belong to the superfamily of ABC transporters. The hydrophobic proteins display a conserved, at least 20 amino acid EAA---G---------I-LP region exposed in the cytosol, the EAA region. We mutagenized the EAA regions of MalF and MalG proteins of the Escherichia coli maltose transport system. Substitutions at the same positions in MalF and MalG have different phenotypes, indicating that EAA regions do not act symmetrically. Mutations in malG or malF that slightly affect or do not affect transport, determine a completely defective phenotype when present together. This suggests that EAA regions of MalF and MalG may interact during transport. Maltose-negative mutants fall into two categories with respect to the cellular localization of the MalK ATPase: in the first, MalK is membrane-bound, as in wild-type strains, while in the second, it is cytosolic, as in strains deleted in the malF and malG genes. From maltose-negative mutants of the two categories, we isolated suppressor mutations within malK that restore transport. They map mainly in the putative helical domain of MalK, suggesting that EAA regions may constitute a recognition site for the ABC ATPase helical domain. PMID- 9214625 TI - Genetic evidence for involvement of type 1, type 2 and type 3 inositol 1,4,5 trisphosphate receptors in signal transduction through the B-cell antigen receptor. AB - Stimulation of B-cell antigen receptor (BCR) induces a rapid increase in cytoplasmic free calcium due to its release from intracellular stores and influx from the extracellular environment. Inositol 1,4,5-trisphosphate receptors (IP3Rs) are ligand-gated channels that release intracellular calcium stores in response to the second messenger, inositol 1,4,5-trisphosphate. Most hematopoietic cells, including B cells, express at least two of the three different types of IP3R. We demonstrate here that B cells in which a single type of IP3R has been deleted still mobilize calcium in response to BCR stimulation, whereas this calcium mobilization is abrogated in B cells lacking all three types of IP3R. Calcium mobilization by a transfected G protein-coupled receptor (muscarinic M1 receptor) was also abolished in only triple-deficient cells. Capacitative Ca2+ entry, stimulated by thapsigargin, remains unaffected by loss of all three types of IP3R. These data establish that IP3Rs are essential and functionally redundant mediators for both BCR- and muscarinic receptor-induced calcium mobilization, but not for thapsigargin-induced Ca2+ influx. We further show that the BCR-induced apoptosis is significantly inhibited by loss of all three types of IP3R, suggesting an important role for Ca2+ in the process of apoptosis. PMID- 9214626 TI - Casein kinase 2 associates with and phosphorylates dishevelled. AB - The dishevelled (dsh) gene of Drosophila melanogaster encodes a phosphoprotein whose phosphorylation state is elevated by Wingless stimulation, suggesting that the phosphorylation of Dsh and the kinase(s) responsible for this phosphorylation are integral parts of the Wg signaling pathway. We found that immunoprecipitated Dsh protein from embryos and from cells in tissue culture is associated with a kinase activity that phosphorylates Dsh in vitro. Purification and peptide sequencing of a 38 kDa protein co-purifying with this kinase activity showed it to be identical to Drosophila Casein Kinase 2 (CK2). Tryptic phosphopeptide mapping indicates that identical peptides are phosphorylated by CK2 in vitro and in vivo, suggesting that CK2 is at least one of the kinases that phosphorylates Dsh. Overexpression of Dfz2, a Wingless receptor, also stimulated phosphorylation of Dsh, Dsh-associated kinase activity, and association of CK2 with Dsh, thus suggesting a role for CK2 in the transduction of the Wg signal. PMID- 9214627 TI - Gi-mediated activation of the Ras/MAP kinase pathway involves a 100 kDa tyrosine phosphorylated Grb2 SH3 binding protein, but not Src nor Shc. AB - Mitogenic G protein-coupled receptors, such as those for lysophosphatidic acid (LPA) and thrombin, activate the Ras/MAP kinase pathway via pertussis toxin (PTX) sensitive Gi, tyrosine kinase activity and recruitment of Grb2, which targets guanine nucleotide exchange activity to Ras. Little is known about the tyrosine phosphorylations involved, although Src activation and Shc phosphorylation are thought to be critical. We find that agonist-induced Src activation in Rat-1 cells is not mediated by Gi and shows no correlation with Ras/MAP kinase activation. Furthermore, LPA-induced tyrosine phosphorylation of Shc is PTX insensitive and Ca2+-dependent in COS cells, but undetectable in Rat-1 cells. Expression of dominant-negative Src or Shc does not affect MAP kinase activation by LPA. Thus, Gi-mediated Ras/MAP kinase activation in fibroblasts and COS cells involves neither Src nor Shc. Instead, we detect a 100 kDa tyrosine phosphorylated protein (p100) that binds to the C-terminal SH3 domain of Grb2 in a strictly Gi- and agonist-dependent manner. Tyrosine kinase inhibitors and wortmannin, a phosphatidylinositol (PI) 3-kinase inhibitor, prevent p100-Grb2 complex formation and MAP kinase activation by LPA. Our results suggest that the p100-Grb2 complex, together with an upstream non-Src tyrosine kinase and PI 3 kinase, couples Gi to Ras/MAP kinase activation, while Src and Shc act in a different pathway. PMID- 9214628 TI - Aberrant axonal projections in mice lacking EphA8 (Eek) tyrosine protein kinase receptors. AB - We have generated mice homozygous for a mutation that disrupts the gene encoding EphA8, a member of the Eph family of tyrosine protein kinase receptors, previously known as Eek. These mice develop to term, are fertile and do not display obvious anatomical or physiological defects. The mouse ephA8/eek gene is expressed primarily in a rostral to caudal gradient in the developing tectum. Axonal tracing experiments have revealed that in these mutant mice, axons from a subpopulation of tectal neurons located in the superficial layers of the superior colliculus do not reach targets located in the contralateral inferior colliculus. Moreover, ephA8/eek null animals display an aberrant ipsilateral axonal tract that projects to the ventral region of the cervical spinal cord. Retrograde labeling revealed that these abnormal projections originate from a small subpopulation of superior colliculus neurons that normally express the ephA8/eek gene. These results suggest that EphA8/Eek receptors play a role in axonal pathfinding during development of the mammalian nervous system. PMID- 9214629 TI - Sympathetic neuron survival and TrkA expression in NT3-deficient mouse embryos. AB - Several in vitro and in vivo studies have led to the widely accepted view that NT3 is required for sympathetic neuroblast survival, induction of TrkA expression and the acquisition of NGF dependence. However, we show that the number of neurons and the levels of trkA and p75 mRNAs in the superior cervical sympathetic ganglion (SCG) of NT3-/- mouse embryos increase normally up to E16, 2 days after SCG neurons start responding to NGF. At E18 and in the postnatal period, there are significant reductions in the number of SCG neurons and in the levels of trkA and p75 mRNAs. These results show that the neurotrophin survival requirements of SCG neurons do not switch from NT3 to NGF during development and that NT3 is not required for the expression of TrkA and p75 and the acquisition of NGF dependence. Rather, some sympathetic neurons have a late requirement for NT3 at the time when they also depend on NGF for survival. The expression of transcripts encoding catalytic TrkC is negligible at this stage, suggesting that NT3 acts mainly via TrkA. PMID- 9214630 TI - Dual targets of a transcriptional activator that tracks on DNA. AB - The sliding clamp of the bacteriophage T4 DNA polymerase, gp45, is also the proximal effector for activation of transcription of T4 late genes. We have identified the phage T4-encoded sigma factor gp55 and the co-activator gp33 as targets of gp45 in promoter complexes, and have shown that a conserved carboxy terminal amino acid sequence of gp55 and gp33 is required for interaction with gp45. The respective contribution of each target-gp45 interaction to activation of transcription has been assessed by measuring promoter opening rates. The opening rate supported by interaction with both targets is far greater than the arithmetical sum of the separate contributions of each target, implying a synergistic activation of transcription through at least two separate interactions of the trimeric gp45. PMID- 9214631 TI - IkappaB alpha is a target for the mitogen-activated 90 kDa ribosomal S6 kinase. AB - The activity of transcription factor NFkappaB is regulated by its subcellular localization. In most cell types, NFkappaB is sequestered in the cytoplasm due to binding of the inhibitory protein IkappaB alpha. Stimulation of cells with a wide variety of agents results in degradation of IkappaB alpha which allows translocation of NFkappaB to the nucleus. Degradation of IkappaB alpha is triggered by phosphorylation of two serine residues, i.e. Ser32 and Ser36, by as yet unknown kinases. Here we report that the mitogen-activated 90 kDa ribosomal S6 kinase (p90rsk1) is an IkappaB alpha kinase. p90rsk1 phosphorylates IkappaB alpha at Ser32 and it physically associates with IkappaB alpha in vivo. Moreover, when the function of p90rsk1 is impaired by expression of a dominant-negative mutant, IkappaB alpha degradation in response to mitogenic stimuli, e.g. 12-O tetradecanoylphorbol 13-acetate (TPA), is inhibited. Finally, NFkappaB cannot be activated by TPA in cell lines that have low levels of p90rsk1. We conclude that p90rsk1 is an essential kinase required for phosphorylation and subsequent degradation of IkappaB alpha in response to mitogens. PMID- 9214632 TI - The LIM-only protein Lmo2 is a bridging molecule assembling an erythroid, DNA binding complex which includes the TAL1, E47, GATA-1 and Ldb1/NLI proteins. AB - The LIM-only protein Lmo2, activated by chromosomal translocations in T-cell leukaemias, is normally expressed in haematopoiesis. It interacts with TAL1 and GATA-1 proteins, but the function of the interaction is unexplained. We now show that in erythroid cells Lmo2 forms a novel DNA-binding complex, with GATA-1, TAL1 and E2A, and the recently identified LIM-binding protein Ldb1/NLI. This oligomeric complex binds to a unique, bipartite DNA motif comprising an E-box, CAGGTG, followed approximately 9 bp downstream by a GATA site. In vivo assembly of the DNA-binding complex requires interaction of all five proteins and establishes a transcriptional transactivating complex. These data demonstrate one function for the LIM-binding protein Ldb1 and establish a function for the LIM only protein Lmo2 as an obligatory component of an oligomeric, DNA-binding complex which may play a role in haematopoiesis. PMID- 9214633 TI - Determinants of chromatin disruption and transcriptional regulation instigated by the thyroid hormone receptor: hormone-regulated chromatin disruption is not sufficient for transcriptional activation. AB - Chromatin disruption and transcriptional activation are both thyroid hormone dependent processes regulated by the heterodimer of thyroid hormone receptor and 9-cis retinoic acid receptor (TR-RXR). In the absence of hormone, TR-RXR binds to nucleosomal DNA, locally disrupts histone-DNA contacts and generates a DNase I hypersensitive site. Chromatin-bound unliganded TR-RXR silences transcription of the Xenopus TRbetaA gene within a canonical nucleosomal array. On addition of hormone, the receptor directs the extensive further disruption of chromatin structure over several hundred base pairs of DNA and activates transcription. We define a domain of the TR protein necessary for directing this extensive hormone dependent chromatin disruption. Particular TR-RXR heterodimers containing mutations in this domain are able to bind both hormone and their thyroid hormone receptor recognition element (TRE) within chromatin, yet are unable to direct the extensive hormone-dependent disruption of chromatin or to activate transcription. We distinguish the hormone-dependent disruption of chromatin and transcriptional activation as independently regulated events through the mutagenesis of basal promoter elements and by altering the position and number of TREs within the TRbetaA promoter. Chromatin disruption alone on a minichromosome is shown to be insufficient for transcriptional activation of the TRbetaA gene. PMID- 9214634 TI - KARP-1: a novel leucine zipper protein expressed from the Ku86 autoantigen locus is implicated in the control of DNA-dependent protein kinase activity. AB - The Ku autoantigen plays an integral role in mammalian DNA double-strand break repair as the DNA binding component of the DNA-dependent protein kinase (DNA-PK) complex. Here, we demonstrate that a second gene, KARP-1 (Ku86 Autoantigen Related Protein-1), is expressed from the Ku86 locus. The KARP-1 gene utilizes an upstream promoter and additional exons which results in an extra 9 kDa of protein appended onto the normal Ku86 polypeptide. The KARP-1-specific domain encodes interdigitating hexa- and penta-heptad repeats of leucine residues flanked by a very basic region. Intriguingly, the catalytic subunit of DNA-PK also contains a hexa-heptad repeat of leucines. Consistent with this observation, we observed that human cell lines stably expressing dominant-negative constructs of KARP-1 resulted in diminished DNA-PK activity and X-ray hypersensitivity and that a KARP 1 antibody significantly neutralized DNA-PK activity in vitro. Finally, we present data which suggests that KARP-1 may be primate-specific. These observations have important repercussions for mammalian DNA double-strand break repair. PMID- 9214635 TI - TTF-2, a new forkhead protein, shows a temporal expression in the developing thyroid which is consistent with a role in controlling the onset of differentiation. AB - Expression of thyroglobulin (Tg) and thyroperoxidase (TPO) genes in thyroid follicular cells occurs in the mouse at embryonic day (E)14.5. Two transcription factors, TTF-1 and Pax-8, have been implicated in transcriptional activation of Tg and TPO, even though the onset of their expression is at E9.5, suggesting that additional events are necessary for transcriptional activation of Tg and TPO genes. We report in this paper the cloning of TTF-2, a DNA binding protein that recognizes sites on both Tg and TPO promoters. TTF-2 is a new forkhead domain containing protein whose expression is restricted to the endodermal lining of the foregut and to the ectoderm that will give rise to the anterior pituitary. TTF-2 shows transient expression in the developing thyroid and anterior pituitary. In the thyroid, TTF-2 expression is down-regulated just before the onset of Tg and TPO gene expression, suggesting that this transcription factor plays the role in development of a negative controller of thyroid-specific gene expression. PMID- 9214636 TI - The HMG-box mitochondrial transcription factor xl-mtTFA binds DNA as a tetramer to activate bidirectional transcription. AB - The mitochondrial HMG-box transcription factor xl-mtTFA activates bidirectional transcription by binding to a site separating two core promoters in Xenopus laevis mitochondrial DNA (mtDNA). Three independent approaches were used to study the higher order structure of xl-mtTFA binding to this site. First, co immunoprecipitation of differentially tagged recombinant mtTFA derivatives established that the protein exists as a multimer. Second, in vitro chemical cross-linking experiments provided evidence of cross-linked dimers, trimers and tetramers of xl-mtTFA. Finally, high resolution scanning transmission electron microscopy (STEM) established that xl-mtTFA binds to the specific promoter proximal site predominantly as a tetramer. Computer analysis of several previously characterized binding sites for xl-mtTFA revealed a fine structure consisting of two half-sites in a symmetrical orientation. The predominant sequence of this dyad symmetry motif shows homology to binding sites of sequence specific HMG-box-containing proteins such as Sry and Lef-1. We suggest that bidirectional activation of transcription results from the fact that binding of a tetramer of xl-mtTFA permits symmetrical interactions with other components of the transcription machinery at the adjacent core promoters. PMID- 9214637 TI - The Pto kinase conferring resistance to tomato bacterial speck disease interacts with proteins that bind a cis-element of pathogenesis-related genes. AB - In tomato, the Pto kinase confers resistance to bacterial speck disease by recognizing the expression of a corresponding avirulence gene, avrPto, in the pathogen Pseudomonas syringae pv. tomato. Using the yeast two-hybrid system, we have identified three genes, Pti4, Pti5 and Pti6, that encode proteins that physically interact with the Pto kinase. Pti4/5/6 each encode a protein with characteristics that are typical of transcription factors and are similar to the tobacco ethylene-responsive element-binding proteins (EREBPs). Using a gel mobility-shift assay, we demonstrate that, similarly to EREBPs, Pti4/5/6 specifically recognize and bind to a DNA sequence that is present in the promoter region of a large number of genes encoding 'pathogenesis-related' (PR) proteins. Expression of several PR genes and a tobacco EREBP gene is specifically enhanced upon Pto-avrPto recognition in tobacco. These observations establish a direct connection between a disease resistance gene and the specific activation of plant defense genes. PMID- 9214639 TI - Variable telomeric repeat synthesis in Paramecium tetraurelia is consistent with misincorporation by telomerase. AB - Telomeric DNA at the ends of chromosomes consist of short, tandem repeat sequences. The telomeres of Paramecium tetraurelia are made up of variable repeats, whereas Paramecium caudatum telomeric repeats are largely invariant. To investigate variable repeat synthesis in P. tetraurelia, mutated telomerase RNA genes were expressed in vivo. We demonstrate that the P. caudatum telomerase RNA can participate in telomere synthesis when expressed in the P. tetraurelia macronucleus, despite 24% primary sequence divergence of the RNAs between the two species. De novo telomeric repeats from transformants indicate that P. tetraurelia telomerase fidelity is dramatically affected by template substitutions and that misincorporation at a single templating position is likely to account for the majority of P. tetraurelia telomeric DNA variability. Furthermore, we show that fidelity is not solely a function of the RNA moiety, as the P. caudatum telomerase RNA does not impart high fidelity to the chimeric enzyme. PMID- 9214638 TI - Mammalian homologues of the Polycomb-group gene Enhancer of zeste mediate gene silencing in Drosophila heterochromatin and at S. cerevisiae telomeres. AB - Gene silencing is required to stably maintain distinct patterns of gene expression during eukaryotic development and has been correlated with the induction of chromatin domains that restrict gene activity. We describe the isolation of human (EZH2) and mouse (Ezh1) homologues of the Drosophila Polycomb group (Pc-G) gene Enhancer of zeste [E(z)], a crucial regulator of homeotic gene expression implicated in the assembly of repressive protein complexes in chromatin. Mammalian homologues of E(z) are encoded by two distinct loci in mouse and man, and the two murine Ezh genes display complementary expression profiles during mouse development. The E(z) gene family reveals a striking functional conservation in mediating gene repression in eukaryotic chromatin: extra gene copies of human EZH2 or Drosophila E(z) in transgenic flies enhance position effect variegation of the heterochromatin-associated white gene, and expression of either human EZH2 or murine Ezh1 restores gene repression in Saccharomyces cerevisiae mutants that are impaired in telomeric silencing. Together, these data provide a functional link between Pc-G-dependent gene repression and inactive chromatin domains, and indicate that silencing mechanism(s) may be broadly conserved in eukaryotes. PMID- 9214640 TI - Localization of Sir2p: the nucleolus as a compartment for silent information regulators. AB - In wild-type budding yeast strains, the proteins encoded by SIR3, SIR4 and RAP1 co-localize with telomeric DNA in a limited number of foci in interphase nuclei. Immunostaining of Sir2p shows that in addition to a punctate staining that coincides with Rap1 foci, Sir2p localizes to a subdomain of the nucleolus. The presence of Sir2p at both the spacer of the rDNA repeat and at telomeres is confirmed by formaldehyde cross-linking and immunoprecipitation with anti-Sir2p antibodies. In strains lacking Sir4p, Sir3p becomes concentrated in the nucleolus, by a pathway requiring SIR2 and UTH4, a gene that regulates life span in yeast. The unexpected nucleolar localization of Sir2p and Sir3p correlates with observed effects of sir mutations on rDNA stability and yeast longevity, defining a new site of action for silent information regulatory factors. PMID- 9214641 TI - Mex67p, a novel factor for nuclear mRNA export, binds to both poly(A)+ RNA and nuclear pores. AB - An essential cellular factor for nuclear mRNA export called Mex67p which has homologous proteins in human and Caenorhabditis elegans was identified through its genetic interaction with nucleoporin Nup85p. In the thermosensitive mex67-5 mutant, poly(A)+ RNA accumulates in intranuclear foci shortly after shift to the restrictive temperature, but NLS-mediated nuclear protein import is not inhibited. In vivo, Mex67p tagged with green fluorescent protein (GFP) is found at the nuclear pores, but mutant mex67-5-GFP accumulates in the cytoplasm. Upon purification of poly(A)+ RNA derived from of UV-irradiated yeast cells, Mex67p, but not nucleoporins Nup85p and Nup57p, was crosslinked to mRNA. In a two-hybrid screen, a putative RNA-binding protein with RNP consensus motifs was found to interact with the Mex67p carboxy-terminal domain. Thus, Mex67p is likely to participate directly in the export of mRNA from the nucleus to the cytoplasm. PMID- 9214642 TI - Mapping metal ions at the catalytic centres of two intron-encoded endonucleases. AB - Divalent metal ions play a crucial role in forming the catalytic centres of DNA endonucleases. Substitution of Mg2+ ions by Fe2+ ions in two archaeal intron encoded homing endonucleases, I-DmoI and I-PorI, yielded functional enzymes and enabled the generation of reactive hydroxyl radicals within the metal ion binding sites. Specific hydroxyl radical-induced cleavage was observed within, and immediately after, two conserved LAGLIDADG motifs in both proteins and at sites at, and near, the scissile phosphates of the corresponding DNA substrates. Titration of Fe2+-containing protein-DNA complexes with Ca2+ ions, which are unable to support endonucleolytic activity, was performed to distinguish between the individual metal ions in the complex. Mutations of single amino acids in this region impaired catalytic activity and caused the preferential loss of a subset of hydroxyl radical cleavages in both the protein and the DNA substrate, suggesting an active role in metal ion coordination for these amino acids. The data indicate that the endonucleases cleave their DNA substrates as monomeric enzymes, and contain a minimum of four divalent metal ions located at or near the catalytic centres of each endonuclease. The metal ions involved in cleaving the coding and the non-coding strand are positioned immediately after the N- and C terminally located LAGLIDADG motifs, respectively. The dual protein/nucleic acid footprinting approach described here is generally applicable to other protein nucleic acid complexes when the natural metal ion can be replaced by Fe2+. PMID- 9214643 TI - Extraplastidic site-specific factors mediate RNA editing in chloroplasts. AB - Single nucleotides in higher plant organellar mRNAs are subject to post transcriptional alterations by RNA editing, typically resulting in changes of the encoded protein sequence. Although some information has been acquired on the general features of the editing processes in both plastids and plant mitochondria, the mechanisms and factors involved in the selective recognition of the nucleotide to be edited are still unknown. To gain a better understanding of how an editing site is specifically selected by the organellar RNA editing machinery, we have attempted to rescue a previously generated tobacco plastid editing mutant. Using an interspecific protoplast fusion approach, we were able to restore RNA editing activity for a specific site in the psbF transcript that otherwise remained unedited. Our results suggest (i) that site-specific trans acting factors mediate chloroplast editing site recognition and (ii) that these factors are of extraplastidic origin. PMID- 9214644 TI - Rules for RNA recognition of GNRA tetraloops deduced by in vitro selection: comparison with in vivo evolution. AB - Terminal loops with a GNRA consensus sequence are a prominent feature of large self-assembling RNA molecules. In order to investigate tertiary interactions involving GNRA loops, we have devised an in vitro selection system derived from a group I ribozyme. Two selections, destined to isolate RNA sequences that would recognize two of the most widespread loops (GUGA and GAAA), yielded variants of previously identified receptors for those loops, and also some yet unrecognized, high-affinity binders with novel specificities towards members of the GNRA family. By taking advantage of available crystal structures, we have attempted to rationalize these results in terms of RNA-RNA contacts and to expose some of the structural principles that govern GNRA loop-mediated tertiary interactions; the role of loop nucleotide 2 in ensuring specific recognition by receptors is emphasized. More generally, comparison of the products of in vitro and natural selection is shown to provide insights into the mechanisms underlying the in vivo evolution of self-assembling RNA molecules. PMID- 9214645 TI - Genome-wide hypermutation in a subpopulation of stationary-phase cells underlies recombination-dependent adaptive mutation. AB - Stationary-phase mutation in microbes can produce selected ('adaptive') mutants preferentially. In one system, this occurs via a distinct, recombination dependent mechanism. Two points of controversy have surrounded these adaptive reversions of an Escherichia coli lac mutation. First, are the mutations directed preferentially to the selected gene in a Lamarckian manner? Second, is the adaptive mutation mechanism specific to the F plasmid replicon carrying lac? We report that lac adaptive mutations are associated with hypermutation in unselected genes, in all replicons in the cell. The associated mutations have a similar sequence spectrum to the adaptive reversions. Thus, the adaptive mutagenesis mechanism is not directed to the lac genes, in a Lamarckian manner, nor to the F' replicon carrying lac. Hypermutation was not found in non revertants exposed to selection. Therefore, the genome-wide hypermutation underlying adaptive mutation occurs in a differentiated subpopulation. The existence of mutable subpopulations in non-growing cells is important in bacterial evolution and could be relevant to the somatic mutations that give rise to cancers in multicellular organisms. PMID- 9214646 TI - The RLF-M component of the replication licensing system forms complexes containing all six MCM/P1 polypeptides. AB - Replication licensing factor (RLF) is involved in preventing re-replication of chromosomal DNA in a single cell cycle, and previously has been separated into two components termed RLF-M and RLF-B. Here we show that Xenopus RLF-M consists of all six members of the MCM/P1 protein family, XMcm2-XMcm7. The six MCM/P1 polypeptides co-eluted on glycerol gradients and gel filtration as complexes with a mol. wt of approximately 400 kDa. In crude Xenopus extract, all six MCM/P1 polypeptides co-precipitated with anti-XMcm3 antibody, although only XMcm5 quantitatively co-precipitated from purified RLF-M. Further fractionation separated RLF-M into two sub-components, one consisting of XMcms 3 and 5, the other consisting of XMcms 2, 4, 6 and 7. Neither of the sub-components provided RLF-M activity. Finally, we show that all six MCM/P1 proteins bind synchronously to chromatin before the onset of S-phase and are displaced as S-phase proceeds. These results strongly suggest that complexes containing all six MCM/P1 proteins are necessary for replication licensing. PMID- 9214647 TI - Licensing of DNA replication by a multi-protein complex of MCM/P1 proteins in Xenopus eggs. AB - In eukaryotes, chromosomal DNA is licensed for a single round of replication in each cell cycle. Xenopus MCM3 protein has been implicated in the licensing of replication in egg extract. We have cloned cDNAs encoding five immunologically distinct proteins associated with Xenopus MCM3 as members of the MCM/P1 family. Six Xenopus MCM proteins formed a physical complex in the egg extract, bound to unreplicated chromatin before the formation of nuclei, and apparently displaced from replicated chromatin. The requirement of six XMCM proteins for the replication activity of the egg extract before nuclear formation suggests that their re-association with replicated chromatin at the end of the mitotic cell cycle is a key step for the licensing of replication. PMID- 9214648 TI - Deletions at stalled replication forks occur by two different pathways. AB - Replication blockage induces non-homologous deletions in Escherichia coli. The mechanism of the formation of these deletions was investigated. A pBR322-mini oriC hybrid plasmid carrying two E. coli replication terminators (Ter sites) in opposite orientations was used. Deletions which remove at least the pBR322 blocking site (named Ter1) occurred at a frequency of 2 x 10(-6) per generation. They fall into two equally large classes: deletions that join sequences with no homology, and others that join sequences of 3-10 bp of homology. Some 95% of the deletions in the former class resulted from the fusion of sequences immediately preceding the two Ter sites, indicating a direct role for blocked replication forks in their formation. These deletions were not found in a topA10 mutant, suggesting a topoisomerase I-mediated process. In contrast, deletions joining short homologous sequences were not affected by the topA10 mutation. However, the incidence of this second class of deletions increased 10-fold in a recD mutant, devoid of exonuclease V activity. This indicates that linear molecules are intermediates in their formation. In addition, approximately 50% of these deletions were clustered in the region flanking the Ter1 site. We propose that they are produced by repair of molecules broken at the blocked replication forks. PMID- 9214649 TI - Second pathway for completion of human DNA base excision-repair: reconstitution with purified proteins and requirement for DNase IV (FEN1). AB - Two forms of DNA base excision-repair (BER) have been observed: a 'short-patch' BER pathway involving replacement of one nucleotide and a 'long-patch' BER pathway with gap-filling of several nucleotides. The latter mode of repair has been investigated using human cell-free extracts or purified proteins. Correction of a regular abasic site in DNA mainly involves incorporation of a single nucleotide, whereas repair patches of two to six nucleotides in length were found after repair of a reduced or oxidized abasic site. Human AP endonuclease, DNA polymerase beta and a DNA ligase (either III or I) were sufficient for the repair of a regular AP site. In contrast, the structure-specific nuclease DNase IV (FEN1) was essential for repair of a reduced AP site, which occurred through the long-patch BER pathway. DNase IV was required for cleavage of a reaction intermediate generated by template strand displacement during gap-filling. XPG, a related nuclease, could not substitute for DNase IV. The long-patch BER pathway was largely dependent on DNA polymerase beta in cell extracts, but the reaction could be reconstituted with either DNA polymerase beta or delta. Efficient repair of gamma-ray-induced oxidized AP sites in plasmid DNA also required DNase IV. PCNA could promote the Pol beta-dependent long-patch pathway by stimulation of DNase IV. PMID- 9214650 TI - Mutation in Escherichia coli under starvation conditions: a new pathway leading to small deletions in strains defective in mismatch correction. AB - Strains of Escherichia coli carrying the mutY mutation lack a mismatch correction glycosylase that removes adenines from various mismatch situations. In growing bacteria, 8-oxoguanine-adenine mispairs persist and can give rise to G-->T transversions during subsequent replication cycles. We now show that when trpA23 mutY bacteria are held under tryptophan starvation conditions the tryptophan independent mutants that arise include small in-frame deletions in addition to transversions. The trpA23 reversion system appears to be unusual in that small in frame deletions occurring in a particular region of the gene can lead to the production of a functional protein. We suggest that this is a consequence of the deletion causing the polar group on the arginine at the trpA23 site to be pulled away from the active site of the enzyme. Such deletions are also found with starved bacteria defective in methyl-directed mismatch correction activity (mutH, mutL or mutS), and deletion mutations are also found among the much lower number of mutants that arise in bacteria wild-type for mismatch correction. There is thus a pathway, hitherto undetected, leading to deletions probably from mismatches under conditions of growth restraint. RecA, UmuC, UvrA, MutH,L,S, SbcC and SbcD proteins are not required for the operation of the deletion pathway. A possible explanation is that the deletion pathway is not dependent upon further replication and that it fails to be discernible in growing cells because it is relatively slow acting and mismatches are likely to encounter a DNA replication fork before the initial step of the deletion pathway. PMID- 9214651 TI - Assembly of a strong promoter following IS911 circularization and the role of circles in transposition. AB - When supplied with high levels of the IS911-encoded transposase, IS911-based transposons can excise as circles in which the right and left terminal inverted repeats are abutted. Formation of the circle junction is shown here to create a promoter, p(junc), which is significantly stronger than the indigenous promoter, pIRL, and is also capable of driving expression of the IS911 transposition proteins. High transposase expression from the circular transposon may promote use of the circle as an integration substrate. The results demonstrate that IS911 circles are highly efficient substrates for insertion into a target molecule in vivo. Insertion leads to the disassembly of p(junc) and thus to a lower level of synthesis of the transposition proteins. The observation that normal levels of IS911 transposition proteins supplied by wild-type copies of IS911 are also capable of generating transposon circles, albeit at a low level, reinforces the idea that the transposon circles might form part of the natural transposition cycle of IS911. These observations form the elements of a feedback control mechanism and have been incorporated into a model describing one possible pathway of IS911 transposition. PMID- 9214652 TI - Relating conformation, Mg2+ binding, and functional group modification in the hammerhead ribozyme. PMID- 9214653 TI - Kissing loops hide premature termination codons in pre-mRNA of selenoprotein genes and in genes containing programmed ribosomal frameshifts. PMID- 9214654 TI - Base dynamics in a UUCG tetraloop RNA hairpin characterized by 15N spin relaxation: correlations with structure and stability. AB - Intramolecular dynamics of guanine and uracil bases in a 14-nt RNA hairpin including the extraordinarily stable UUCG tetraloop were studied by 15N spin relaxation experiments that are sensitive to structural fluctuations occurring on a time scale of picoseconds to nanoseconds. The relaxation data were interpreted in the framework of the anisotropic model-free formalism, using assumed values for the chemical shift anisotropies of the 15N spins. The rotational diffusion tensor was determined to be symmetric with an axial ratio of 1.34 +/- 0.12, in agreement with estimates based on the ratio of the principal moments of the inertia tensor. The model-free results indicate that the bases of the G x U pair in the tetraloop are at least as rigid as the interior base pairs in the stem, whereas the 5'-terminal guanine is more flexible. The observed range of order parameters corresponds to base fluctuations of 19-22 degrees about the chi torsion angle. The results reveal dynamical consequences of the unusual structural features in the UUCG tetraloop and offer insights into the configurational entropy of hairpin formation. PMID- 9214655 TI - Naegleria nucleolar introns contain two group I ribozymes with different functions in RNA splicing and processing. AB - We have characterized the structural organization and catalytic properties of the large nucleolar group I introns (NaSSU1) of the different Naegleria species N. jamiesoni, N. andersoni, N. italica, and N. gruberi. NaSSU1 consists of three distinct RNA domains: an open reading frame encoding a homing-type endonuclease, and a small group I ribozyme (NaGIR1) inserted into the P6 loop of a second group I ribozyme (NaGIR2). The two ribozymes have different functions in RNA splicing and processing. NaGIR1 is an unusual self-cleaving group I ribozyme responsible for intron processing at two internal sites (IPS1 and IPS2), both close to the 5' end of the open reading frame. This processing is hypothesized to lead to formation of a messenger RNA for the endonuclease. Structurally, NaGIR2 is a typical group IC1 ribozyme, catalyzing intron excision and exon ligation reactions. NaGIR2 is responsible for circularization of the excised intron, a reaction that generates full-length RNA circles of wild-type intron. Although it is only distantly related in primary sequence, NaSSU1 RNA has a predicted organization and function very similar to that of the mobile group I intron DiSSU1 of Didymium, the only other group I intron known to encode two ribozymes. We propose that these twin-ribozyme introns define a distinct category of group I introns with a conserved structural organization and function. PMID- 9214656 TI - Transfer RNA recognition by the Escherichia coli delta2-isopentenyl pyrophosphate:tRNA delta2-isopentenyl transferase: dependence on the anticodon arm structure. AB - To elucidate the sequence elements required in the anticodon stem for the recognition of Escherichia coli tRNA(Ser) (GGA) by the E. coli isopentenyl tRNA:A37 transferase (IPTT), which result in the conversion of A37 into isopentenylated i6A37, we have tested and characterized in vitro T7-runoff transcripts of 17 variants of E. coli tRNA(Ser)(GGA) and 7 other tRNAs from E. coli and yeast. Our results indicate that, instead of a stringent specific anticodon stem and loop sequence, the key feature required for the recognition of E. coli tRNAs by IPTT is the A36A37A38 sequence occurring within the seven membered anticodon loop, and the retention of the standard helical structure and flexibility, especially in the proximal anticodon stem. The G30*U40 mismatch base pair close to the anticodon loop is strictly avoided. The frequent occurrence of a C-G base pair in the three stem locations closest to the loop (positions 29-41, 30-40 and 31-39) or the occurrence of even one such C-G base pair along with some other similarly less suited, but individually tolerated deviations can also totally abolish the A37 isopentenylation of tRNA. For the position 30-40, the G-C base pair is shown uniquely suited, whereas for the adjoining 29-41 stem location, a purine-pyrimidine base pair with pyrimidine on the 3'-side is strongly preferred. Retention of the overall 3D tRNA structure is favorable for isopentenylation and allows some tolerance of proximal stem sequence deviations. Our data suggest a recognition mode that implies the interaction of IPTT with the strictly conserved A36A37A38 sequence and the other functional groups located in the minor groove of the anticodon stem. PMID- 9214657 TI - In vitro optimization of truncated stem-loop II variants of the hammerhead ribozyme for cleavage in low concentrations of magnesium under non-turnover conditions. AB - By truncating helix II to two base pairs in a hammerhead ribozyme having long flanking sequences (greater than 30 bases), the rate of cleavage in 1 mM magnesium can be increased roughly 100-fold. Replacing most of the nucleotides in a typical stem-loop II with 1-4 randomized nucleotides gave an RNA library that, even before selection, was more active in 1 mM magnesium than the parent ribozyme, but considerably less active than the truncated stem-loop II ribozyme. A novel, multiround selection for intermolecular cleavage was exploited to optimize this library for cleavage in low concentrations of magnesium. After three rounds of selection at sequentially lower concentrations of magnesium, the library cleaved substrate RNA 20-fold faster than the initial pool and was cloned. This pool was heavily enriched for one particular sequence (5'-CGUG-3') that represented 16 of 52 isolates (the next most common sequence was represented only six times). This sequence also represented the most active sequence, exceeding the activity of the short helix II variant under the conditions of the selection, thereby demonstrating the effectiveness of the selection technique. Analysis of the cleavage rates of RNAs made from eight isolates having different four-base insert sequences allowed assignment of highly preferred bases at each position in the insert. Analysis of pool clones having insert of differing lengths showed that, in general, activity decreased as the length of the insert decreased from 4 to 1. This supports the suggested role of stem-loop II in stabilizing the non-Watson-Crick interactions between the conserved bases of the catalytic core. PMID- 9214658 TI - Mutational analysis of the protein subunits of the signal recognition particle Alu-domain. AB - Two polypeptides of the murine signal recognition particle (SRP), SRP9 and SRP14, bind exclusively as a heterodimer to SRP RNA and their presence is required for elongation arrest activity of the particle. SRP9/14 also constitute a subunit of small cytoplasmic Alu RNPs. To identify RNA-binding determinants, we assayed the dimerization and RNA-binding capacities of altered proteins in vitro. Despite the structural homology of the two proteins, their requirements for dimerization differ substantially. In SRP9, an internal fragment of 43 amino acids is sufficient to allow dimer formation, whereas in SRP14 only few changes, such as removing an internal loop region, are tolerated without affecting its dimerization activity. The dimerization defect of the SRP14 proteins is most likely explained by a reduced stability or ability to fold of the proteins. Interestingly, SRP RNA can engage certain dimerization-defective SRP14 proteins into stable complexes, suggesting that low-affinity interactions between the RNA and SRP14 may help to overcome the folding defect or the reduced stability of the proteins. We identified two regions, one in each protein, that are essential for RNA-binding. In SRP9, acidic amino acid residues in the N-terminal alpha-helix and the adjacent loop and, in SRP14, a flexible internal loop region are critical for RNA-binding. In the heterodimer, the two regions are located in close proximity, consistent with the RNA-binding region being formed by both proteins. PMID- 9214659 TI - Mutation of PTB binding sites causes misregulation of alternative 3' splice site selection in vivo. AB - Alternative splicing of pre-mRNA is a commonly used mechanism to regulate gene expression in higher eukaryotes. However, with the exception of regulated cascades in Drosophila, the cis-acting elements and the trans-acting factors that control tissue- and/or developmentally regulated splicing remain largely unidentified. Cis-acting elements that control smooth muscle-specific repression of exon 3 of alpha-tropomyosin (alpha-TM) have been identified recently and consist of two regions that flank this exon. Deletion of either element causes misregulated splicing of alpha-TM in transfected smooth muscle cells. In experiments designed to characterize essential sequences within each element and the factors that interact with these sequences, we have identified two overlapping sequences within the downstream regulatory element (DRE) that are identical to binding sites for polypyrimidine tract binding protein (PTB) that were identified using iterative selection techniques. Mutation of these sites caused aberrant splicing regulation in transfected smooth muscle cells. In addition, sequences identical to high-affinity PTB binding sites were also detected upstream of exon 3 and mutation of these sites also resulted in misregulation of splicing in vivo, suggesting that PTB binding to specific sequences flanking exon 3 is responsible, in part, for the repression of exon 3. Consistent with this hypothesis, UV crosslinking and equilibrium binding assays confirm that the same mutations that cause misregulated splicing also disrupt PTB binding to RNA. PMID- 9214660 TI - Secondary structure of the 3'-noncoding region of flavivirus genomes: comparative analysis of base pairing probabilities. AB - The prediction of the complete matrix of base pairing probabilities was applied to the 3' noncoding region (NCR) of flavivirus genomes. This approach identifies not only well-defined secondary structure elements, but also regions of high structural flexibility. Flaviviruses, many of which are important human pathogens, have a common genomic organization, but exhibit a significant degree of RNA sequence diversity in the functionally important 3'-NCR. We demonstrate the presence of secondary structures shared by all flaviviruses, as well as structural features that are characteristic for groups of viruses within the genus reflecting the established classification scheme. The significance of most of the predicted structures is corroborated by compensatory mutations. The availability of infectious clones for several flaviviruses will allow the assessment of these structural elements in processes of the viral life cycle, such as replication and assembly. PMID- 9214661 TI - Global similarities in nucleotide base composition among disparate functional classes of single-stranded RNA imply adaptive evolutionary convergence. AB - The number of distinct functional classes of single-stranded RNAs (ssRNAs) and the number of sequences representing them are substantial and continue to increase. Organizing this data in an evolutionary context is essential, yet traditional comparative sequence analyses require that homologous sites can be identified. This prevents comparative analysis between sequences of different functional classes that share no site-to-site sequence similarity. Analysis within a single evolutionary lineage also limits evolutionary inference because shared ancestry confounds properties of molecular structure and function that are historically contingent with those that are imposed for biophysical reasons. Here, we apply a method of comparative analysis to ssRNAs that is not restricted to homologous sequences, and therefore enables comparison between distantly related or unrelated sequences, minimizing the effects of shared ancestry. This method is based on statistical similarities in nucleotide base composition among different functional classes of ssRNAs. In order to denote base composition unambiguously, we have calculated the fraction G+A and G+U content, in addition to the more commonly used fraction G+C content. These three parameters define RNA composition space, which we have visualized using interactive graphics software. We have examined the distribution of nucleotide composition from 15 distinct functional classes of ssRNAs from organisms spanning the universal phylogenetic tree and artificial ribozymes evolved in vitro. Surprisingly, these distributions are biased consistently in G+A and G+U content, both within and between functional classes, regardless of the more variable G+C content. Additionally, an analysis of the base composition of secondary structural elements indicates that paired and unpaired nucleotides, known to have different evolutionary rates, also have significantly different compositional biases. These universal compositional biases observed among ssRNAs sharing little or no sequence similarity suggest, contrary to current understanding, that base composition biases constitute a convergent adaptation among a wide variety of molecular functions. PMID- 9214662 TI - A new strategy for introducing photoactivatable 4-thiouridine ((4S)U) into specific positions in a long RNA molecule. AB - We describe a new protocol, which does not require (4S)UpG, for introducing (4S)U into specific sites in a pre-mRNA substrate. A 5'-half and a full-length RNA are first synthesized by phage RNA polymerase. p(4S)Up, which is derived from (4S)UpU and can therefore be 32P-labeled, is then ligated to the 3' end of the 5'-half RNA with T4 RNA ligase. The 3' phosphate of the ligated product is removed subsequently by CIP (calf intestinal alkaline phosphatase) to produce a 3'-OH group. The 3'-half RNA with a 5' phosphate is produced by site-specific RNase H cleavage of the full-length pre-mRNA directed by a 2'-O-methyl RNA-DNA chimera. The two half RNAs are then aligned with a bridging oligonucleotide and ligated with T4 DNA ligase. Our results show that 32P-p(4S)Up ligation to the 3' end of the 5'-half RNA is comparable to 32P-pCp ligation. Also, the efficiency of the bridging oligonucleotide-mediated two-piece ligation is quite high, approximately 30-50%. This strategy has been applied to the P120 pre-mRNA containing an AT-AC intron, but should be applicable to many other RNAs. PMID- 9214663 TI - Multidrug resistance in breast cancer--is the jury in yet? PMID- 9214664 TI - Meta-analysis under the microscope. PMID- 9214666 TI - Faux chromosomes hit the streets, hold real promise for gene therapy. PMID- 9214667 TI - A gene by any other name: does whimsy matter? PMID- 9214665 TI - Ovulation, p53 mutations, and ovarian cancer--a causal link? PMID- 9214668 TI - Treatment for rare brain tumors veers from traditional tack. PMID- 9214669 TI - Next generation of SERMs being seen in clinic. PMID- 9214670 TI - President's cancer panel probes pervasive link between race and health outcomes. PMID- 9214671 TI - Multidrug resistance in breast cancer: a meta-analysis of MDR1/gp170 expression and its possible functional significance. AB - BACKGROUND: P-glycoprotein (gp170; encoded by the MDR1 gene [also known as PGY1]) is a membrane protein capable of exporting a variety of anticancer drugs from cells. MDR1/gp170 expression has been studied in breast cancer, but the prevalence of this expression and its role in breast tumor drug resistance are unclear. PURPOSE: We conducted a critical review and meta-analysis of studies examining MDR1/gp170 expression in breast cancer to estimate the likely prevalence and clinical relevance of this expression. We also explored reasons for differences in the findings from individual studies. METHODS: Published papers on MDR1/gp170 expression in breast cancer were identified by searching several literature databases and reviewing the bibliographies of identified papers. Variability across the studies in the proportion of tumors expressing MDR1/gp170 was assessed by use of chi-squared tests of homogeneity, weighted means, and weighted linear regression. Pooled relative risks (RRs) for the association between the induction of MDR1/gp170 expression and prior chemotherapy and associations between MDR1/gp170 expression and several clinical outcomes were estimated by use of Mantel-Haenszel methods. Heterogeneity among the pooled RRs was explored by use of chi-squared tests. Reported P values are two-sided. RESULTS: Thirty-one studies were identified and evaluated. The proportion of breast tumors expressing MDR1/gp170 in all of the studies was 41.2%, but there was substantial heterogeneity in the values across individual studies (P<.0001). Regression analyses demonstrated that a considerable portion of the observed heterogeneity was a consequence of the change, over time, from RNA hybridization based assays to immunohistochemistry-based assays of MDR1/gp170 expression. Measuring MDR1/gp170 expression before versus after chemotherapy and use of cytotoxic drugs that are not substrates for gp170 also contributed to the heterogeneity. Treatment with chemotherapeutic drugs or hormonal agents was associated with an increase in the proportion of tumors expressing MDR1/gp170 (RR = 1.77; 95% confidence interval [CI] = 1.46-2.15). Patients with tumors expressing MDR1/gp170 were three times more likely to fail to respond to chemotherapy than patients whose tumors were MDR1/gp170 negative (RR = 3.21; 95% CI = 2.28-4.51); this RR increased to 4.19 (95% CI = 2.71-6.47) when considering only patients whose tumor expression of MDR1/gp170 was measured after chemotherapy. MDR1/gp170 expression was not associated with lymph node metastases, estrogen receptor status, tumor size, tumor grade, or tumor histology. CONCLUSIONS AND IMPLICATIONS: MDR1/gp170 expression in breast tumors is associated with treatment and with a poor response to chemotherapy. The data are consistent with a contributory role for MDR1/gp170 in the multidrug resistance in some breast tumors. PMID- 9214672 TI - Relationship between lifetime ovulatory cycles and overexpression of mutant p53 in epithelial ovarian cancer. AB - BACKGROUND: Several lines of evidence have suggested a relationship between a woman's number of ovulatory cycles and the development of ovarian epithelial cancer. Repair of the ovarian surface after ovulation requires cellular proliferation, and spontaneous mutations arising during the DNA synthesis that accompanies this proliferation may play a role in carcinogenesis. PURPOSE: We conducted a molecular epidemiologic study to test the hypothesis that a greater number of ovulatory cycles increases the risk of ovarian cancer by inducing proliferation-associated DNA damage. In particular, we examined the association between the lifetime number of ovulatory cycles and mutation of the p53 tumor suppressor gene (also known as TP53) in ovarian tumors. METHODS: Case-case and case-control analyses involving participants in the Cancer and Steroid Hormone study were used to examine the association between p53 gene mutation in ovarian tumors and the lifetime number of ovulatory cycles. The women in our study were 20-54 years of age and included 197 case patients with invasive ovarian epithelial cancer and 3363 control subjects. Mutation of the p53 gene was indicated by overexpression of p53 protein (i.e., cellular accumulation of mutant p53 protein) in paraffin-embedded ovarian cancer tissue blocks; the mutant protein was detected by means of standard immunohistochemical techniques. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by employing multivariate analyses, with the use of logistic regression. Reported P values are two-sided. RESULTS: Women whose cancers overexpressed p53 protein (p53 positive) had a greater mean number of lifetime ovulatory cycles (388 +/- 77.4 cycles [mean +/- standard deviation]) than women whose cancers did not overexpress p53 protein (p53 negative) (342 +/- 119.0 cycles) (P = .0025). Furthermore, women with p53 positive tumors were more likely to have had moderate (i.e., 235-375) or high (i.e., 376-533) numbers of ovulatory cycles than women with p53-negative tumors (age-adjusted ORs = 7.0 [95% CI = 1.6-30.5] and 7.7 [95% CI = 1.4-41.2], respectively) (< or = 234 cycles was the referent category). After controlling for age, menopausal status, and nulliparity, women with p53-positive tumors were found to be significantly more likely to have had moderate or high numbers of ovulatory cycles than control subjects (ORs = 4.3 [95% CI = 1.4-13.0] and 9.1 [95% CI = 2.7-30.9], respectively); the corresponding ORs for women with p53 negative tumors compared with control subjects were 0.6 (95% CI = 0.3-1.4) and 1.3 (95% CI = 0.5-3.2), respectively. CONCLUSIONS AND IMPLICATIONS: A higher number of ovulatory cycles may be associated with increased amounts of proliferation-associated DNA damage and increased risk of developing p53-positive but not p53-negative epithelial ovarian cancer. Our results are consistent with more than one developmental pathway in the pathogenesis of this type of cancer. PMID- 9214673 TI - Birth characteristics, sibling patterns, and acute leukemia risk in childhood: a population-based cohort study. AB - BACKGROUND: The occurrence of acute lymphoblastic leukemia (ALL) and acute myeloid leukemia (AML) during childhood may be influenced by factors operating in fetal life. Furthermore, childhood ALL has been suggested to be linked to patterns of infection during infancy. PURPOSE: To explore these hypotheses and other associations, we studied the impact of sibling patterns (e.g., birth order) and birth characteristics (e.g., birth weight) on the risk of childhood ALL and AML. METHODS: By linkage of records of population-based registries, a cohort of all children whose mothers were born in Denmark from April 1935 through March 1978 was established. Children who developed ALL or AML during the period from April 1968 through December 1992 were identified by linkage with the Danish Cancer Registry. Birth weights were obtained for children born during the period from January 1973 through December 1992 by linkage with the Medical Birth Registry. RESULTS: The cohort of approximately 2.0 million children was followed for the diagnosis of ALL or AML for 20.9 million person-years. A total of 704 cases of childhood ALL were identified. Among 0-4 year olds, the relative risks (RRs) of ALL for birth order positions 1, 2, 3, and 4+ were 1.00 (reference), 0.85 (95% confidence interval [CI] = 0.68-1.07), 0.91 (95% CI = 0.66-1.25), and 0.57 (95% CI = 0.30-1.06), respectively (P for trend = .09). A decreasing trend was not observed among 5-14 year olds. A significant log-linear association between birth weight and the risk of ALL was observed for both age groups. Overall, the RR of ALL increased by a factor of 1.46 (95% CI = 1.18-1.81) (P = .0005) for each kilogram of increase in birth weight. A total of 114 cases of childhood AML were identified. Children born second or later in the birth order had an increased risk of AML (RR = 1.53; 95% CI = 1.01-2.32) compared with firstborns. A particularly high risk of AML at ages 2 (RR = 2.53; 95% CI = 1.46 4.40) and 3 years was associated with having siblings compared with being an only child at those ages. Similar to the findings for ALL risk, there was a significant association between birth weight and AML risk. The relative increase in AML risk per 1-kg increase in birth weight was 2.14 (95% CI = 1.19-3.85; P = .009). CONCLUSION AND IMPLICATIONS: The association between birth weight and childhood leukemia suggests the importance of intrauterine factors. A plausible explanation may be that increasing birth weight is associated with a higher rate of cell proliferation and/or a larger number of precursor cells being at risk of malignant transformation. The inverse association between birth order and ALL risk among 0-4 year olds was weak, but it was compatible with the hypothesis that delayed exposure to infection may increase the risk of ALL in this age group. The association of childhood AML with birth order and sibship size at young ages deserves further attention in the search for environmental factors that affect childhood AML risk. PMID- 9214674 TI - Leisure-time physical activity, body size, and colon cancer in women. Nurses' Health Study Research Group. AB - BACKGROUND: Physical inactivity and high body mass index (weight in kilograms divided by height in square meters) have been linked to increased risk of colon cancer. However, none of the few prospective studies in women has shown a statistically significant reduction in colon cancer incidence or mortality associated with increased leisure-time physical activity. PURPOSE: In this prospective study, we asked whether leisure-time physical activity, body mass index, or body fat distribution could significantly influence the risk of colon cancer in women. METHODS: The participants in this study were enrolled in the Nurses' Health Study, which began in 1976. Every 2 years, the women provided additional personal information and information on medical risk factors and major medical events. The time spent per week at a variety of leisure-time physical activities was determined, and the time spent at each activity was multiplied by its typical energy expenditure, expressed in terms of metabolic equivalents or METs. The resulting values for each woman were added to yield an MET-hours-per week score. Reported diagnoses of colon cancer were confirmed by review of hospital records and pathology reports. Relative risks and associated 95% confidence intervals were calculated. RESULTS: In multivariate analyses that included body mass index, women who expended more than 21 MET-hours per week on leisure-time physical activity had a relative risk of colon cancer of 0.54 (95% confidence interval [CI] = 0.33-0.90) in comparison with women who expended less than 2 MET-hours per week. Women who had a body mass index greater than 29 kg/m2 had a relative risk of colon cancer of 1.45 (95% CI = 1.02-2.07) in comparison with women who had a body mass index less than 21 kg/m2. A tendency toward higher colon cancer risk was observed for increasing waist-to-hip ratio (relative risk = 1.48 [95% CI = 0.88-2.49] for comparison of the highest quintile ratio [>0.833] to the lowest [<0.728]). CONCLUSIONS AND IMPLICATIONS: The significant inverse association between leisure-time physical activity and incidence of colon cancer in women in this study is consistent with what has been found in men. Recommendations to increase physical activity and maintain lean body weight should receive greater emphasis as part of a feasible approach to the prevention of colon cancer. PMID- 9214675 TI - Prostate cancer susceptibility locus on chromosome 1q: a confirmatory study. AB - BACKGROUND: Recent recognition that a predisposition to prostate cancer can be inherited has led to a search for specific genes associated with the disease. Through a study of families with three or more affected first-degree relatives, a region on the long arm of chromosome 1 (i.e., 1q24-25) has been tentatively identified as containing a gene, HPC1, involved in the development of hereditary prostate cancer. Confirmation of this finding is needed, however, before attempts are made to isolate and characterize the putative HPC1 gene. PURPOSE: To confirm that chromosome 1q24-25 contains a gene relevant to hereditary prostate cancer, we analyzed an independent set of families, each with two or more affected individuals. METHODS: Fifty-nine unrelated families were selected for analysis on the sole criterion that more than one living family member was affected by prostate cancer. DNA samples were subsequently isolated from 130 individuals with the disease. These samples were genotyped at six polymorphic marker sequences (D1S215, D1S2883, D1S466, D1S158, D1S518, and D1S2757) covering the chromosomal region proposed to contain HPC1. The resulting data were analyzed by nonparametric multipoint linkage (NPL) methods, yielding NPL Z scores and corresponding one-sided P values. RESULTS: When the entire set of 59 families was considered, the occurrence of prostate cancer (and, presumably, the HPC1 gene) was most tightly linked to marker D1S466 (NPL Z score = 1.58; P = .0574). Analysis of the 20 families (51 affected individuals) fulfilling one or more of the proposed clinical criteria for hereditary prostate cancer (i.e., three or more affected individuals within one nuclear family; affected individuals in three successive generations [maternal or paternal lineage]; and/or clustering of two or more individuals affected before the age of 55 years) revealed more convincing evidence of disease linkage to chromosome 1q24-25 (maximum NPL Z score [at marker D1S466] = 1.72; P = .0451). The 39 families (79 affected individuals) that did not meet the clinical criteria for hereditary prostate cancer exhibited no significant evidence of disease linkage to DNA sequences at chromosome 1q24-25 (maximum NPL Z score [at marker D1S466] = 0.809; P = .208). The six African American families in our study contributed disproportionately to the observation of linkage, with a maximum NPL Z score at marker D1S158 of 1.39 (P = .0848) for these families. CONCLUSIONS AND IMPLICATIONS: Our data confirm that chromosome 1q24-25 is likely to contain a prostate cancer susceptibility gene. Future efforts at positional cloning of the HPC1 gene should focus on families who meet the proposed clinical criteria for hereditary prostate cancer. PMID- 9214676 TI - Regional differences in known risk factors and the higher incidence of breast cancer in San Francisco. AB - BACKGROUND: The age-adjusted incidence of breast cancer in the San Francisco Bay Area has consistently been higher than that in other regions of the United States. The distribution of established risk factors for breast cancer (i.e., parity, age at first full-term pregnancy, breast-feeding, age at menarche, and age at menopause) and probable risk factors (e.g., alcohol consumption) also differs across geographic regions. PURPOSE: A study was planned to explore the extent to which differences in the regional distribution of established and probable risk factors could explain the increased incidence of breast cancer in the San Francisco Bay Area. METHODS: Age-adjusted breast cancer incidence rates for January 1978 through December 1982 were obtained for the San Francisco Bay Area and other regions from the Surveillance, Epidemiology, and End Results (SEER) Program. Risk factor data from January 1980 through December 1982 were computed from the Cancer and Steroid Hormone Study, a population-based, case control study of women 22-55 years of age who resided in eight SEER regions. Two different statistical methods were used to compute the relative risk (RR) of breast cancer associated with residence in the San Francisco Bay Area versus other regions, after adjusting for regional differences in known risk factors. RESULTS: Substantial differences in the distribution of breast cancer risk factors were found between the San Francisco Bay Area and other regions. Nearly all of these differences would be expected to lead to an elevated incidence of breast cancer in the San Francisco Bay Area. With the use of incidence rates adjusted only for age, the RR for San Francisco Bay Area residence from January 1978 through December 1982 compared with residence in seven other SEER areas was 1.14 for white women and 1.10 for black women. Depending on the statistical method used, the RR was reduced to approximately 0.96-0.99 for white women and 0.75-0.83 for black women, after further adjusting for established and probable risk factors (parity, age at first full-term pregnancy, breast-feeding, age at menarche, age at menopause, and alcohol consumption). Without adjustment for alcohol consumption, the corresponding results were 0.97-1.02 for white women and 0.77-0.88 for black women. CONCLUSIONS: Among both white women and black women, the elevated breast cancer incidence rate in the San Francisco Bay Area can be completely accounted for by regional differences in known risk factors. PMID- 9214677 TI - Re: prediction of carboplatin clearance from standard morphological and biological patient characteristics. PMID- 9214678 TI - Re: you say tomato and I say tomahto: getting a handle on pronouncing apoptosis. PMID- 9214679 TI - von Hippel-Lindau disease gene alterations associated with endolymphatic sac tumor. PMID- 9214680 TI - Inhibition of mammary carcinogenesis in rats by parenteral high-dose vitamin E. PMID- 9214681 TI - Antipsoriatic and proinflammatory action of anthralin. Implications for the role of oxygen radicals. AB - Anthralin is among the most effective agents for the topical treatment of psoriasis. However, this drug causes unpleasant side-effects such as inflammation and staining of the nonaffected skin surrounding a psoriatic lesion. The biochemical basis for the induction of an inflammatory response in the skin and the antipsoriatic effectiveness are uncertain, although several cellular targets of anthralin action have been identified. Because no single mechanism is operative, the view was taken that all the effects exerted by anthralin are caused by its redox activity leading to the generation of anthralin free radicals and oxygen radicals. Clear relationships between oxygen-radical production by anthralin and biological response are evident with respect to chemical lesions in cellular macromolecules such as DNA, lipid membranes, and enzymes, indicating that these species account for the antipsoriatic and proinflammatory effects elicited by anthralin. This poses new challenges for the medicinal chemist and provides impetus for identifying novel compounds having potential for an improved therapeutic index. PMID- 9214682 TI - Interaction of oligonucleotide-conjugates with the dipeptide transporter system in Caco-2 cells. AB - Oligonucleotide-based therapies represent novel strategies for manipulating the expression and function of target proteins and are undergoing clinical evaluation for the treatment of viral diseases and malignancies. However, poor biological stability and cellular delivery represent potential limitations to the therapeutic development of oligonucleotides. Conjugation of oligonucleotides to lipophilic groups can improve delivery to cells but the enhanced cellular binding may also facilitate nonspecific interactions. In this report, we show that phosphorothioate oligonucleotides conjugated to lipophilic groups, either tocopherol (Vitamin E) or 2-Di-O-hexadecyl-3-glycerol, can significantly inhibit the functioning of the dipeptide transporter system (DTS) in cultured Caco-2 intestinal cells. Because the DTS mediates the binding and absorption of nutrient peptides and important drugs, such as the cephalosporin and penicillin antibiotics, this finding has important implications in relation to the potential toxicity of lipophilic conjugates in vivo. It also suggests a potential drug interaction with lipophilic oligonucleotide-conjugates if they were to be delivered orally. PMID- 9214683 TI - Effects of nucleosomes and anti-tumor drugs on the catalytic activity of type II DNA topoisomerase from rat testis. AB - To gain insight into the relative catalytic efficiencies of mammalian type I and type II DNA topoisomerases, in the cellular context, we have used naked DNA and DNA incorporated into nucleosomes as substrates. We observed that the relaxation activity of both the enzymes declined with DNA containing increasing densities of nucleosomes; however, kinetic analysis revealed that topoisomerase I seemed less affected than topoisomerase II. The addition of histone H1, in stoichiometric amounts, to naked DNA or minichromosomes lessened the activity of topoisomerase II, and required 7-fold less for complete inhibition when the latter was used as the substrate. To ascertain if the observed differences are specific to topoisomerase II from testis, we examined the effect of nucleosomes on the catalytic efficiency of its isoform from liver. Interestingly, the suppression of relaxation activity of liver topoisomerase II required substrates containing higher mass ratios of histone octamer/DNA. Studies on the effect of nucleosomes on the action of teniposide displayed significant differences in the kinetics of the reaction, in its IC50 values, and have provided biochemical evidence for the first time that nucleosomes increased inhibition caused by teniposide. Further, this feature appears to be specific for topoisomerase II-directed drugs and is not shared by the generic class of either DNA-intercalating or non-DNA intercalating ligands. PMID- 9214684 TI - Effect of protein kinase inhibitors on activity of mammalian small heat-shock protein (HSP25) kinase. AB - The aim of this study was to investigate different protein kinase inhibitors (secondary metabolite-derived substances, synthetic compounds, and substrate based peptides) for their potency to inhibit the mammalian small heat shock protein (HSP25) kinase (E.C. 2.7.1.37) isolated from Ehrlich ascites tumor cells. Among the secondary metabolite-derived inhibitors (staurosporine, K-252a, K-252b, KT5926, KT5720, erbstatin analog, and quercetin) and synthetic compounds (H-9, H 89, HA 1004, KN-62, ML-7, tyrphostin A25, and tyrphostin B42), KT5926, staurosporine, and K-252a inhibited HSP25 kinase most efficiently. Kinetic analysis revealed that inhibition by staurosporine (Ki = 32.4 nM) and K-252a (Ki = 13.7 nM) was competitive with ATP. Inhibition by KT5926 was competitive with the substrate peptide KKKALNRQLSVAA (Ki = 27.2 nM) and noncompetitive with respect to ATP (Ki = 38.8 nM). In comparison with other protein kinases, HSP25 kinase was relatively resistant to most of the inhibitors. KT5926 was the only tested inhibitor with certain preference for HSP25 kinase when compared with protein kinases A, C, and G. Among the tested substrate-based peptides, we identified one peptide (KKKALNRQLGVAA), which preferentially inhibited HSP25 kinase in comparison with protein kinases A and C and mitogen-activated protein kinase. This peptide inhibited HSP25 kinase competitively with the substrate peptide (Ki = 8.1 microM) and noncompetitively with ATP (Ki = 134 microM). A peptide (SRVLKEDKERWEDVK) derived from the putative autoinhibitory domain of the closely related human mitogen-activated protein kinase-activated protein kinase-2 did not inhibit HSP25 kinase activity, suggesting the existence of several species of HSP25 kinases. Furthermore, the data identified structural requirements for inhibitors of HSP25-kinase. PMID- 9214685 TI - Detection of EP2, EP4, and FP receptors in human ciliary epithelial and ciliary muscle cells. AB - We have examined the expression of three prostaglandin (PG) receptors, EP2, EP4, and FP, in a nonpigmented ciliary epithelial cell line (ODMCl-2) and in human ciliary muscle (HCM) cells. Total RNA preparations from either ODMCl-2 or HCM cells were subjected to reverse transcription-polymerase chain reaction (RT-PCR) with sense and antisense primers for each of the three PG receptors. The RT-PCR generated DNA products of predicted sizes corresponding to the EP2, EP4, and FP receptors in both ODMCl-2 and HCM cells. PCR products corresponding to each receptor were hybridized with specific 32P-labeled probes and, for further confirmation, digested with appropriate restriction enzymes. Pharmacological studies with the EP2 receptor-selective agonist butaprost resulted in a significant increase in the cyclic AMP level in ODMCl-2 cells. The stimulation of cyclic AMP in ODMCl-2 cells by PGE2 and 11-deoxy PGE1, the respective EP1/EP2/EP3/EP4 and EP2/EP3/EP4 receptor agonists, was concentration-dependently inhibited by the EP4 receptor-selective antagonist AH23848. These results conclusively demonstrate the presence of both mRNA and protein for EP2, EP4, and FP receptors in ODMCl-2 and HCM cells. PMID- 9214686 TI - Human platelet activation is inhibited upstream of the activation of phospholipase A2 by U73343. AB - U73122 is known as an inhibitor of phospholipase C (PLC; EC 3.1.4.11). Its close structural analogue, U73343, lacks this activity and is used as a control compound. We have found that both compounds interfere with platelet signal transduction. U73122 completely abolished aggregation evoked by thrombin, TG, and collagen. Aggregation evoked by TG and collagen was also blocked by U73343, an effect due to inhibition of TxA2 production. U73343 was a potent inhibitor of TG evoked arachidonic acid release, but a weak inhibitor of cytosolic phospholipase A2 (cPLA2; EC 3.1.1.4) activity. Cytosolic PLA2 activation in platelets involves protein tyrosine phosphorylation. U73343 inhibited TG- and collagen-evoked protein tyrosine phosphorylation, which can thus explain its action against these agents. These data indicate that caution is needed when using U73343 along with U73122 in the study of intracellular signalling pathways. PMID- 9214687 TI - Effects of R(-)-1-(benzo[b]thiophen-5-yl)-2-[2-N,N-diethylamino)ethoxy]ethan ol hydrochloride (T-588), a novel cognitive enhancer, on noradrenaline release in rat cerebral cortical slices. AB - We investigated the effects of R(-)-1-(benzo[b]thiophen-5-yl)-2-[2-(N,N diethylamino)ethoxy]ethan ol hydrochloride (T-588), a novel cognitive enhancer, on noradrenaline (NA) release from rat cerebral cortical slices in vitro. Addition of T-588 in an assay mixture stimulated [3H]NA release from prelabeled slices in the presence or absence of extracellular CaCl2, and in the presence of the Ca2+/calmodulin antagonists N-(6-aminohexyl)-5-chloro-1 naphthalenesulfonamide and trifluoperazine. T-588 stimulated NA release with a time lag of about 1 min, and the high level of release was maintained for at least 10 min, whereas maximal KCl-evoked NA release was observed within 1 min after the addition of KCl, and the effect declined subsequently. The effect of T 588 was reversible (pretreatment with T-588 showed no effect on NA release after two washes by centrifugation). We also compared the effects of T-588 and N ethylmaleimide (NEM), a sulfhydryl alkylating agent known to stimulate neurotransmitter release in several types of cells. The addition of NEM stimulated NA release irreversibly from the slices in a Ca2+-independent manner, and the effect of NEM, but not that of T-588, was inhibited by the simultaneous addition of dithiothreitol, a sulfhydryl group reducing agent. The addition of T 588, which stimulated NA release by itself, inhibited the NA release by 0.6 mM NEM, although the effect of T-588 was additive in the presence of 0.2 mM NEM. These findings suggest that T-588 stimulates NA release from rat cerebral cortical slices in a Ca2+- and calmodulin-independent manner, possibly via an NEM sensitive factor(s), although the mechanism of the effects of T-588 seems to be different from that of NEM. PMID- 9214688 TI - Modulation of cyclic AMP levels in a clonal neural cell line by inhibitors of tyrosine phosphorylation. AB - The convergence of tyrosine kinase and cyclic AMP (cAMP) signal transduction pathways was investigated in the HT4.7 neural cell line with inhibitors of tyrosine kinases and tyrosine phosphatases. The protein tyrosine kinase inhibitor genistein inhibited isoproterenol-stimulated cAMP production by 40-60% in whole cells, with no effect on basal cAMP levels. In both whole cells and membranes, genistein also inhibited cAMP produced in response to direct stimulation of adenylyl cyclase with forskolin. However, in the absence of phosphodiesterase inhibitors, genistein presentation resulted in an increase in cAMP levels. Genistein inhibited phosphodiesterase activity by 80-85%, indicating that tyrosine phosphorylation stimulates both cAMP synthesis and degradation. The decrease in cAMP levels by genistein was not merely competitive inhibition of adenylyl cyclase with respect to ATP, since the Km of adenylyl cyclase for ATP remained essentially the same in either the presence or the absence of genistein. Another tyrosine kinase inhibitor, herbimycin A, which inhibits by a different mechanism than genistein, also decreased forskolin-stimulated cAMP in whole cells. As would be expected for the involvement of tyrosine phosphorylation in the control of cAMP production, inhibition of tyrosine phosphatases by vandate increased forskolin-stimulated cAMP production. These results suggest that cAMP production can be regulated by tyrosine phosphorylation, and the simultaneous activation of both cAMP synthesis and degradation may serve to alter the duration of cAMP elevation. PMID- 9214689 TI - Enzymic denitrosation of 1,3-dimethyl-2-cyano-1-nitrosoguanidine in rat liver cytosol and the fate of the immediate product S-nitrosoglutathione. AB - The tumorigenicity of certain N-nitrosoguanidinium compounds is limited, in rodents, by the propensity of these agents to be detoxified by denitrosation. Previous studies have revealed that rodent glutathione transferase isoenzymes are capable of catalyzing this process, generating exclusively the denitrosated guanidinium compound and S-nitrosoglutathione (GSNO). Experiments considering the denitrosation of 1,3-dimethyl-2-cyano-1-nitrosoguanidine (CyanoDMNG) in rat liver cytosol incubates are reported, with emphasis on the fate of GSNO. Incubates composed with equimolar CyanoDMNG and reduced glutathione (GSH) effected 100% denitrosation; the GSNO yield was less than expected as was the quantity of GSH consumed. When the anticipated 100% yield concentration of GSNO was applied to cytosol incubates, 20-40% of it rapidly disappeared. Nitrosated protein thiols accounted for 35% of the NO moiety released, nitrite ion 30%, and nitric oxide production was detectable. Concomitant with GSNO loss, GSH and oxidized glutathione (GSSG) were generated in yields similar to those detected in the CyanoDMNG/GSH incubates. Thus, the fate of GSNO in cytosol determines the yields of glutathione-based products, and the stoichiometry of the glutathione transferase reaction is demonstrated. In incubates composed with equimolar CyanoDMNG, GSH, and NADPH, denitrosation was again 100%, but GSNO yields were very low and residual GSH increased. Inclusion of NADPH in incubates containing the anticipated 100% yield concentration of GSNO resulted in rapid GSNO degradation, producing GSH and a detected but unidentified product; S-nitrosated protein, nitrite, and nitrate yields were minimal, nitric oxide production was abolished, and incubate response to a mercuric chloride/azo dye assay approached zero. The fate of the NO moiety consequent to this GSNO catabolism is presently unknown. PMID- 9214690 TI - Denitrosation of 1,3-dimethyl-2-cyano-1-nitrosoguanidine in rat primary hepatocyte cultures. AB - N-Nitrosoguanidines are potential carcinogens. However, the toxicity of these agents is attenuated significantly in laboratory rodents by processes that remove the nitroso group to generate the relatively innocuous parent guanidinium compound. The denitrosation of 1,3-dimethyl-2-cyano-1-nitrosoguanidine (CyanoDMNG) mediated by rat hepatocytes in primary culture was investigated. At concentrations < or = 200 microM, applied CyanoDMNG was converted efficiently to 1,3-dimethyl-2-cyanoguanidine (CyanoDMG). In trials using 50 microM CyanoDMNG (5 mL dosing solutions), it was demonstrated that hepatocytes are capable of denitrosating a 40 microM concentration of the applied compound with little change in the total or oxidized glutathione levels. The process was inhibited by coincidently applied ethacrynic acid, a glutathione transferase inhibitor. Reduction of hepatocyte glutathione to 20% of control levels by buthionine sulfoximine pretreatment had little effect on CyanoDMG production; total depletion of cytosolic glutathione by diethyl maleate pretreatment arrested CyanoDMNG processing. Hepatocyte-mediated CyanoDMNG denitrosation did not generate nitrite; nitrate yields were 10% relative to the CyanoDMG produced. The mercuric chloride/azo dye response of cultures lysed at times during 50 microM CyanoDMNG processing indicated intact CyanoDMNG as the only dye-sensitive material present. At applied CyanoDMNG > 100 microM, S-nitrosoglutathione (GSNO) yields were detectable; 4 microM GSNO was generated (concentration in 5 mL lysates) and maintained during 60 min at the 200 microM CyanoDMNG treatment level; this yield decayed if CyanoDMNG was withdrawn. Based on these and previous findings, it is hypothesized that CyanoDMNG is converted to CyanoDMG and GSNO by glutathione transferases and that GSNO is catabolized to eventually regenerate reduced glutathione. The fate of most of the NO moiety released remains to be determined. PMID- 9214691 TI - Activation of protein kinase C by 1,4-dihydro-2,6-dimethyl-5-nitro-4-[2 (trifluoromethyl)-phenyl]-3-py rid ine carboxylic acid methyl ester (Bay K 8644), a calcium channel agonist, in alveolar type II cells. AB - A role for calcium channels in the regulation of surfactant secretion is suggested by the observation that endothelin-1-stimulated surfactant secretion is inhibited by calcium channel blockers. 1,4-Dihydro-2,6-dimethyl-5-nitro-4-[2 (trifluoromethyl)-phenyl]-3-pyridi ne carboxylic acid methyl ester (Bay K 8644), a dihydropyridine derivative, stimulates voltage-dependent and non-voltage dependent calcium channels in a number of cell types. This study demonstrates that Bay K 8644 increased phosphatidylcholine (PC) secretion in isolated lung epithelial type II cells in a time- and concentration-dependent manner with an EC50 of 100 +/- 8 nM (mean +/- SEM, N = 6). The secretagogue effect of Bay K 8644 was independently decreased in the absence of external calcium, or in the presence of nifedipine, a calcium channel antagonist, or inhibitors of protein kinase C (PKC). Bay K 8644 increased intracellular calcium from 130 +/- 8 to 230 +/- 14 nM (N = 6, P < 0.05), an effect that was blocked by nifedipine. Bay K 8644 also increased the membrane-associated PKC activity in a concentration-dependent manner. In the membranes from Bay K 8644-stimulated cells, the increase in calcium-dependent PKC was greater than that in the calcium-independent PKC, suggesting preferential translocation of calcium-dependent PKC to the membranes. We suggest that both elevated calcium and activation of PKC are required for calcium agonist Bay K 8644-induced surfactant secretion in type II cells. PMID- 9214692 TI - Inhibition of thymocyte apoptosis by berberine. AB - To find anti-apoptotic substances in plant resources, a microassay method for estimating DNA fragmentation was established using fluorochrome 3,5 diaminobenzoic acid dihydrochloride. Examination was made of various herbal medicines for inhibitory effects on glucocorticoid-induced apoptosis in thymocytes. Several Kampo medicines, e.g. Oren-gedoku-to and San'o-shashin-to, were found to inhibit dexamethasone-induced apoptosis in murine thymocytes. Some of these medicines contain Coptidis rhizoma (CR) as the major constituent, and the CR extract showed the most potent inhibitory activity on thymocyte apoptosis of more than 200 species of herbal extracts. The inhibition of apoptosis by CR extract was confirmed by the trypan blue exclusion test, lactate dehydrogenase release measurement, and morphological evaluation by electron microscopy. The benzodioxolo-benzoquinolizine alkaloid, berberine, and five berberine-type alkaloids, isolated from CR extract, had an inhibitory effect, whereas no effect was noted for the aporphin-type alkaloid magnoflorine. The inhibitory action of berberine was also demonstrated on etoposide- and camptothecin-induced apoptosis. PMID- 9214693 TI - Compartmentalisation and characteristics of a Ca2+-dependent phospholipase A2 in human colon mucosa. AB - The biochemical properties of the phospholipase A2 (PLA2) found in the 100,000 x g centrifugate cytosol or particulate fractions of human colonic mucosa have been investigated using both deoxycholate-solubilized and Escherichia coli (E. coli) phospholipids as substrates. PLA2 activity was present in both subcellular fractions and the profiles of biochemical activites were similar. Activity in the particulate fraction was approximately twofold greater than the cytosol fraction when expressed on the basis of protein concentration. The PLA2 is Ca2+ dependent and using EGTA-regulated buffers cytosolic or particulate fraction activity was similar at both 10 microm or 10 mm Ca2+ concentrations. Using deoxycholate phospholipid micelles as substrate a small but statistically significant twofold preference for glycero-phosphatidylcholine bearing sn-2-arachidonate compared with sn-2-oleate was seen, but this preference was not noted using arachidonate or oleate labelled E. coli membranes. Dithiothreitol (10 mM) reduced colon mucosal cytosol PLA2 activity significantly by 63.5 +/- 1.90% in cytosol and by 30.54 +/- 1.27% in microsomes using micelles as substrate or by 84.3 +/- 2.30% in cytosol and by 69.33 +/- 11.30% in microsomes using oleate-labelled E. coli as substrates. Warming at 57 degrees C reduced activity significantly by 35.0 +/- 5.80% in microsomes and by 40.0 +/- 7.08% in cytosol. Acid treatment increased PLA2 activity to 148 +/- 16.3% in microsomes and 145 +/- 18.6% in cytosol. When mucosal preparations were subjected to heparin-Sepharose chromatography, it bound tightly and eluted in the same position on a salt gradient as authentic human group II PLA2. Further purification by gel-permeation chromatography gave activity in the 14 kDa region of the elution profile. These features have many of the characteristics expected of a 14 kDa isoform of PLA2 but exhibit activity at concentrations of Ca2+ that are relevant in the intracellular environment and may participate in cellular lipid metabolism. PMID- 9214694 TI - Leukocyte function-associated antigen 1-dependent adhesion of rat hepatoma AH66F cells and inhibition by protein kinase C inhibitors. AB - When rat ascites hepatoma AH66F cells were incubated on a mesothelial cell (M cell) layer for 1 hr, the adhesion rate of the cells to M-cells was ca. 46%. The protein kinase C (PKC) inhibitors, N-(2-methylpiperazyl)-5 isoquinolinesulfonamide (H-7) and N-ethoxycarbonyl-7-oxostaurosporine (NA-382), inhibited the adhesion of AH66F cells in a concentration-dependent manner, and the effect of NA-382 appeared after a treatment of more than 24 hr. The decreased adhesion rate after treatment with NA-382 for 48 hr was not further inhibited by addition of monoclonal antibodies of leukocyte function-associated antigen-1 (LFA 1) alpha- and beta-chains and intercellular adhesion molecule-1 (ICAM-1) (WT.1, WT.3, and 1A29, respectively). The expression of LFA- 1 alpha- and beta-chains on the surface of the plasma membrane of AH66F cells was decreased after treatment with NA-382 for 48 hr; treatment with a potent inhibitor of cyclic AMP-dependent protein kinase, N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide (H 89), did not affect the cell adhesion and the expression of LFA-1 molecules on AH66F cells. These results suggest that the expression of LFA-1 molecules on AH66F cells is regulated through the PKC pathway. PMID- 9214695 TI - Sinusoidal endothelial cells as a target for acetaminophen toxicity. Direct action versus requirement for hepatocyte activation in different mouse strains. AB - Hepatic congestion occurs early in acetaminophen poisoning. This study examines whether acetaminophen is toxic to sinusoidal endothelial cells (SEC), which might lead to microcirculatory disruption. Acetaminophen toxicity was examined in vivo and in vitro in SEC and hepatocytes from C3H-HEN and Swiss Webster mice. In both strains, there was significantly more toxicity to SEC than to hepatocytes; in SEC from C3H-HEN mice, acetaminophen was directly toxic, but the presence of hepatocytes was required for toxicity to Swiss SEC. Acetaminophen, 750 mg/kg, by gavage caused toxicity with variability within and between strains, but all animals died between 3.5 and 6 hr with zone 3 hemorrhagic necrosis. Pretreatment of C3H-HEN SEC with aminobenzotriazole, a suicide inhibitor of P450, abolished toxicity. Baseline glutathione (GSH) levels were comparable, but a 12-hr incubation with acetaminophen decreased GSH by 60 and 8%, respectively, in C3H HEN and Swiss SEC in single cell type culture. In co-culture, under conditions where Swiss SEC viability declined by 73%, hepatocyte viability and GSH only decreased by 21 and 20%, respectively. In conclusion, acetaminophen was toxic to SEC. It was directly toxic to SEC in one mouse strain and required hepatocyte activation in another strain. The lack of direct toxicity to Swiss SEC may be due to the lack of an activating P450 isozyme. Zone 3 hemorrhagic necrosis in vivo was comparable in both strains, despite differences in the pathways leading to SEC toxicity in vitro. We propose that toxicity to SEC may contribute to hepatic congestion in acetaminophen intoxication. PMID- 9214696 TI - Resveratrol inhibits metal ion-dependent and independent peroxidation of porcine low-density lipoproteins. AB - Resveratrol, a phytoalexin (3, 4', 5, trihydroxystilbene) present in some red wines, has been reported to inhibit copper-mediated low-density lipoprotein (LDL) oxidation. In this study, we examined the efficiency of this compound in inhibiting metal ion-dependent and independent peroxidation of porcine LDL. At 0.5, 1, or 1.5 microM, transresveratrol prolonged the lag time preceding the onset of conjugated diene formation in a dose-dependent manner, with a slope of the propagation phase 5-fold greater in the presence of Cu SO4 (5 microM) than in the presence of the free radical generator, AAPH [2, 2'-azobis (2-amidinopropane) dihydrochloride] (1 mM). At 1 microM, transresveratrol prolonged the lag time 3.4 and 1.4-fold in the presence of copper and AAPH, respectively. Isomerisation into cisresveratrol significantly lowered the chelating capacity, but did not alter the free radical scavenging capacity. As compared to flavonoids and trolox, transresveratrol showed a much higher ability to prolong the lag time in copper, but not in AAPH-catalyzed oxidation. The kinetics of generation of degradative products in the presence of copper confirmed the strongest protective effects of transresveratrol, because the formation of thiobarbituric acid reactive substances and hydroperoxides was almost completely inhibited at 200 min. By contrast, transresveratrol was less potent than flavonoids (but more than trolox) as a scavenger of free radicals. Our data show that, like flavonoids, resveratrol protects LDL against peroxidative degradation by both chelating and free radical scavenging mechanisms. However, transresveratrol, which is by far the most potent chelator of copper, does not chelate iron. It might contribute to the protective effects of wine polyphenols by removing copper from LDL particles and arterial tissue and, thereby, delaying the consumption of flavonoids and endogenous antioxidants. PMID- 9214698 TI - Evidence against peroxisome proliferation-induced hepatic oxidative damage. AB - It has been proposed that nongenotoxic peroxisome proliferators may cause hepatocellular cancer by an oxidative damage-mediated mechanism(s). The argument for this hypothesis is based mainly on the noted ability of peroxisome proliferators to induce significantly H2O2-producing peroxisomal beta-oxidation while causing a minimal induction of H2O2-degrading catalase. The recent discovery, accurate determination, and use of isoprostanes as a sensitive indicator of oxidative damage prompted us to investigate whether induction of hepatic peroxisomal beta-oxidation in male B6C3F1 mice is accompanied by elevated levels of isoprostanes in those livers. The data show that while 7 days of feeding mice a diet containing 100 ppm [4-chloro-6-(2,3-xylidino)-2 pyrimidinylthio]acetic acid (WY-14,643) increased peroxisomal beta-oxidation by 16-fold and catalase activity by only 2-fold, hepatic levels of esterified F2 isoprostanes were not altered. These levels were 2.8 +/- 0.5 ng/g liver in control mice and 2.4 +/- 0.1 ng/g liver in mice fed the experimental diet for 7 days. Consequently, it is concluded that oxidative stress does not appear to occur in response to peroxisome proliferation, as evidenced by the lack of increase in hepatic levels of F2-isoprostanes in livers of mice treated with the potent peroxisome proliferator WY-14,643. PMID- 9214697 TI - Apoptotic death in adenocarcinoma cell lines induced by butyrate and other histone deacetylase inhibitors. AB - n-Butyrate inhibits the growth of colon cancer cell lines. In the HCT 116 cell line, butyrate-induced growth inhibition is almost fully reversible, whereas in the VACO 5 cell line, a subpopulation undergoes apoptosis within 30 hr of treatment with butyrate. Concurrent treatment of VACO 5 cells with butyrate and the phorbol ester 12-O-tetradecanoylphorbol 13-acetate (TPA) accelerates and increases the incidence of cell death to nearly 100% of the population, whereas HCT 116 cells largely remain alive during treatment with this combination. The action of butyrate as an inhibitor of histone deacetylase was assessed in these cell lines by examining extracted core histones for their electrophoretic mobility in Triton/acid/urea gels. The concentrations of butyrate that were effective for inducing apoptosis were similar to the concentrations that caused hyperacetylation of core histones in the VACO 5 cell line. Furthermore, an examination of other carboxylic acids for induction of apoptosis revealed a rank order that corresponded to the order of potency in causing hyperacetylation of core histones. Specifically, the active acids were 3-5 carbons in length and lacked substitution at the 2-position. Isovaleric and propionic acids, in particular, proved to be effective inducers of both hyperacetylation and apoptosis at 5 mM concentrations, a finding of potential relevance to the unusual pancytopenia occurring after acidotic episodes in isovaleric and propionic acidemias. The duration of butyrate treatment required for chromatin fragmentation (10-20 hr) corresponded to the time required for histone H4 to become predominantly tetraacetylated. Furthermore, trichostatin A, a structurally dissimilar inhibitor of histone deacetylase, mimicked butyrate-induced apoptosis of VACO 5 cells and growth inhibition of HCT 116 cells. The dramatic enhancement of VACO 5 cell death by TPA, and the high level resistance of HCT 116 cells to butyrate were not evident from histone acetylation determinations. Thus, applications of butyrate for cytoreduction therapy will benefit from pharmacodynamic assessment of histone acetylation, but will require additional work to predict susceptibility to butyrate-induced death. PMID- 9214699 TI - Cardiac microdialysis of salicylic acid .OH generation on nonenzymatic oxidation by norepinephrine in rat heart. AB - The effect of pargyline, a monoamine oxidase inhibitor, on the generation of hydroxyl free radicals (.OH) was investigated using cardiac microdialysis. Salicylic acid in Ringer's solution (0.5 nmol x microL(-1) x min(-1)) was infused directly through a microdialysis probe to detect the generation of .OH as reflected by the formation of dihydroxybenzoic acid (DHBA) in the myocardium of anesthetized rats. When pargyline (100 nmol x microL(-1) x min(-1)) was infused in rat heart, the level of norepinephrine (NE) gradually increased in a time dependent manner and an increase of DHBA was also observed. When NE was administered to the pargyline pretreated animals, a marked elevation in the levels of 2,3- and 2,5-DHBA formation was obtained, as compared to the group treated with NE only, showing a positive linear correlation between NE and .OH formation trapped as 2,3-DHBA (R2 = 0.981) or 2,5-DHBA (R2 = 0.984) in the dialysate. NE clearly produced an increase in .OH formation. These results indicate that accumulation of NE in the extracellular fluid elicited by pargyline can be auto-oxidized, which in turn, leads (possibly by an indirect mechanism) to the formation of cytotoxic .OH free radicals. PMID- 9214700 TI - Differential regulation of expression of rat hippocampal muscarinic receptor subtypes following fimbria-fornix lesion. AB - Quantitative RNase protection assays were performed to determine the levels of muscarinic receptor subtype (m1-m5) mRNAs in rat hippocampi. Results showed that the m1, m3, and m4 subtype mRNAs were expressed at relatively high levels, but the levels of the m2 and m5 subtype were very low. Three weeks following aspiration lesions of the fimbria-fornix to produce cholinergic denervation of the hippocampus, non-M1 receptors (non-pirenzepine displaceable [3H]quinuclindinyl benzilate binding sites) in the hippocampus were increased significantly, which correlated with increases in the levels of hippocampal m3 and m4 receptor mRNAs (m3: +24% and m4: +41%). These findings indicate that multiple muscarinic receptor subtypes are expressed in the hippocampus with the m3 and m4 subtypes predominantly postsynaptic to the septohippocampal cholinergic terminals. PMID- 9214701 TI - Induction by interleukin-1 (IL-1) of the mRNA of histidine decarboxylase, the histamine-forming enzyme, in the lung of mice in vivo and the effect of actinomycin D. AB - It is known that the activity of histidine decarboxylase (HDC), the histamine forming enzyme, is induced in response to various stimuli. However, it has repeatedly been reported that actinomycin D (Act D), a typical inhibitor of RNA synthesis, is either ineffective, or actually potentiates induction of this enzyme. Thus, it has been suggested that the induction of HDC may not require the formation of mRNA, i.e. that pre-formed, long-lived mRNA molecules may be responsible for the induction. In the present study, we examined the effects of interleukin-1alpha (IL-1alpha) on the amount of HDC mRNA present during the induction of HDC activity. In mice injected with IL-1alpha, HDC mRNA increased in the lung, spleen and stomach, but was hardly detectable in these tissues in control (saline-injected) mice. In the lung, the time course of the rise and fall in HDC mRNA was shorter than that of the rise and fall in HDC activity. In the present study, actinomycin D (Act D) did not inhibit the increase in HDC mRNA induced by IL-1alpha; in fact, it potentiated the elevation of both HDC mRNA and HDC activity. These results suggest that IL-1alpha induces HDC activity or its enzyme protein through the formation of short-lived HDC mRNA molecules. This is the first demonstration that Act D can enhance an increase in HDC mRNA: this potentiating, rather than inhibiting, effect is discussed. PMID- 9214702 TI - 1997 USPHS/IDSA guidelines for the prevention of opportunistic infections in persons infected with human immunodeficiency virus. USPHS/IDSA Prevention of Opportunistic Infections Working Group. PMID- 9214703 TI - Studies on chemical modification of monensin. VI. Preparation of C-7 modified monensins via protected 7-oxomonensin and evaluation of their ion transport activity. AB - 7-Carboxymethylmonensin (3), 7-aminomonensin (4), and 7-oxomonensin (5) were synthesized via protected 7-oxomonensin (2), which is expected to be a useful intermediate for the preparation of various monensin derivatives. intermediate for the preparation of various monensin derivatives. Compounds 3-5 exhibited smaller ion transport activity than monensin (1). PMID- 9214704 TI - Oxygenation reaction of methyl valproate with a mono-oxygenase model reagent: implication for stereochemical identification of the mammalian metabolites of valproic acid. AB - Non-enzymic oxygenation reaction of methyl valproate (2) utilizing a simple model system for mono-oxygenases, Fe(MeCN)2(6+)-H2O2-Ac2O in MeCN, was investigated in connection with stereochemical analyses of the mammalian metabolites of 1. This oxygenation reaction of methyl valproate (2) gave a 92:8 mixture of the anti isomer 4a and the syn-isomer 4b, together with 5a, and 5b corresponding to the mammalian metabolites of 1. The stereochemistry of 4a, 5a, and 5b was elucidated by spectral analyses of the corresponding beta-lactone 6a, gamma-lactone 7a and 7b prepared from the oxygenation products. The asymmetric synthesis of (+)-7a was also achieved. PMID- 9214705 TI - Studies on anti-inflammatory agents. IV. Synthesis and pharmacological properties of 1,5-diarylpyrazoles and related derivatives. AB - A series of novel 1,5-diarylpyrazole derivatives was synthesized and tested for anti-inflammatory and analgesic activities to develop anti-inflammatory agents with fewer side effects than existing nonsteroidal anti-inflammatory drugs. The structure-activity relationships in this series were extensively studied. Electron-withdrawing substituents such as CN and CF3 were optimal at the 3 position of the pyrazole ring. Replacement of these substituents with bulky ones gave less active compounds. The 4-(methylsulfonyl)phenyl group seemed to be the optimal group at the 5-position of the pyrazole ring. The most potent compound was 1-(4-fluorophenyl)-5-[4-(methylsulfonyl)phenyl]-pyrazole-3-carbonitrile (19a), with oral ED50 value of 0.030 and 0.47 mg/kg on adjuvant-induced arthritis and collagen-induced arthritis, respectively, and an ED30 value of 7.4 mg/kg in the yeast-induced hyperalgesia (Randall-Selitto) assay. Compound 19a also showed potent inducible cyclooxygenase (COX-2)-inhibitory activity (IC50 = 0.24 microM) with no COX-1 inhibition even at 100 microM. PMID- 9214706 TI - Synthesis of novel succinamide derivatives having the 5,11-dihydro-6H-pyrido[2,3 b][1,4]benzodiazepin-6-one skeleton as potent and selective M2 muscarinic receptor antagonists. I. AB - A series of 5,11-dihydro-6H-pyrido[2,3-b][1,4]benzodiazepin-6-one derivatives containing the succinamide skeleton has been synthesized and evaluated for M1, M2 and M3 muscarinic receptor binding affinities (in vitro) and M2 and M3 muscarinic receptor antagonistic activities (in vivo). Some of them showed higher and more selective binding affinities for M(2) muscarinic receptors than that of AF-DX 116. Among them, 11-[3-[N-[2-(N-benzyl-N- methylamino)ethyl]-N ethylcarbamoyl]propionyl]-5,11-dihydro-6H-pyr ido [2,3-b][1,4]benzodiazepin-6-one (68) was found to be the most potent and selective M2 muscarinic receptor antagonist in vitro. This compound also strongly inhibited the oxotremorine induced bradycardia after intravenous administration and showed 130-fold selectivity for M2 muscarinic receptors over M3 muscarinic receptors in vivo. PMID- 9214707 TI - Synthesis biological evaluation of alkyl, alkoxy, alkylthio, or amino-substituted 2,3-dihydro-1,5-benzothiazepin-4(5H)-ones. AB - 2,3-Dihydro-1,5-benzothiazepin-4(5H)-ones substituted with an alkyl, alkoxy, alkylthio, hydroxy, or amino group on the fused benzene ring of the 1,5 benzothiazepine skeleton were synthesized and their vasodilating, antihypertensive, and platelet aggregation-inhibitory activities were investigated. (-)-cis-3-Acetoxy-5-[2-(di-methylamino) ethyl]-2,3-dihydro-8-methyl 2-(4-methylphenyl)-1,5-benzothiazepin- 4(5H)-one ((-)-13e) was selected for further studies as a potent inhibitor of platelet aggregation. PMID- 9214708 TI - Nicotinamide derivatives as a new class of gastric (H+/K+)-ATPase inhibitors II. Synthesis and structure-activity relationships of 2[(2,4 dimethoxybenzyl)sulfinyl]-N-(4-pyridinyl)pyridine-3-carboxamides. AB - Members of a new series of 2-[(2,4-dimethoxybenzyl)sulfinyl]-N-(4 pyridinyl)pyridine-3-carboxamides were synthesized and evaluated for their gastric antisecretory activity and the ability to inhibit cytochrome P450 dependent O-dealkylation of 7-ethoxycoumarin (7-EC) in rat liver microsomes. Several of the compounds synthesized exhibited potent inhibitory activities against both [14C]aminopyrine accumulation stimulated by dibutyryl cyclic AMP in isolated rabbit parietal cells and histamine-induced gastric acid secretion in pylorus-ligated rats when administered intraduodenally; their inhibitory activities were equivalent to or superior to those of the parent compound [2- [(2,4-dimethoxybenzyl)sulfinyl]-N-(4-pyridinyl)pyridine-3-carboxamide] and omeprazole. Among the compounds having potent antisecretory activity in vitro and in vivo, 2-[(2,4-dimethoxybenzyl)sulfinyl]-N-(2,5-dimethyl-4-pyridinyl) pyridine 3-carboxamide and 2-[(2,4-dimethoxybenzyl)sulfinyl]-N-(2,6-dimethyl-4 pyridinyl)pyridine-3 - carboxamide in particular showed lower inhibitory activity against the 7-EC deethylase than omeprazole. It seems probable that, unlike omeprazole, these compounds do not interact with a metabolism of other drugs in vivo. These compounds, therefore, are considered to be more promising candidate agents for treating acid-related gastrointestinal disorders than the parent compound reported previously. PMID- 9214709 TI - Bioactive saponins and glycosides. VIII. Notoginseng (1): new dammarane-type triterpene oligoglycosides, notoginsenosides-A, -B, -C, and -D, from the dried root of Panax notoginseng (Burk.) F.H. Chen. AB - The glycosidic fraction from the dried roots of Panax notoginseng (Burk.) F.H. Chen was found to show protective effect on liver injury induced by D galactosamine and lipopolysaccharide. From the glycosidic fraction with hepatoprotective effect, nine new dammarane-type triterpene oligoglycosides, notoginsenosides-A, -B, -C, -D, -E, -G, -H, -I, and -J and an acetylenic fatty acid glycoside, notoginsenic acid beta-sophoroside, were isolated together with fourteen known dammarane-type triterpene oligoglycosides. The structures of notoginsenosides-A, -B, -C and -D were determined on the basis of chemical and physicochemical evidence, which included the chemical correlation with ginsenoside-Rb1 using photosensitized oxygenation, as follows: notoginsenoside A; 3-O-[beta-D-glucopyranosyl (1-->2)-beta-D-glucopyranosyl]-20-O-[beta-D glucopyranosyl (1-->6)-beta-D-glucopyranosyl] 3 beta, 12 beta,20(S),25 tetrahydroxydammar -23-ene; B; 3-O-[beta-D-glucopyranosyl (1-->2)-beta-D glucopyranosyl]-20-O-[beta-D-glucopyranosyl (1-->6)-beta-D- glucopyranosyl] 3 beta, 12 beta,20(S)-trihydroxydammar-25-en-24-one, C; 3-O-[beta-D-glucopyranosyl (1-->2)-beta-D-glucopyranosyl]-20-O-[beta-D -glucopyranosyl (1-->6)-beta-D glucopyranosyl] 3 beta,12 beta,20(S)- trihydroxy-24 zeta-hydroperoxydammar-25 ene, and D; 3-O-[beta-D-xylopyranosyl (1-->2)-beta-D-glucopyranosyl (1-->2)-beta D-glucopyranosyl]-20-O-[beta-D-xylopyranosyl (1-->6)-beta-D-glucopyranosyl (1- >6)-beta-D-glucopyranosyl]20(S)-protopanaxadiol, respectively. PMID- 9214710 TI - gamma-Pyrones from Gonystylus keithii, as new inhibitors of parathyroid hormone (PTH)-induced Ca release from neonatal mouse calvaria. AB - New gamma-pyrones, 9'-oxopodopyrone (3) and 8-methyl-9'-oxopodopyrone (4) were isolated from the leaves of Gonystylus keithii, along with known gamma-pyrones, 10'-oxopodopyrone (1) and 8-methyl-10'-oxopodopyrone (2). These gamma-pyrones markedly inhibited the bovine parathyroid hormone (PTH)-induced Ca release from neonatal mouse calvaria in vitro. It is the first time that gamma-pyrones showed inhibitory effects on bone resorption, and these compounds may be seed compounds of new drugs for osteoporosis. PMID- 9214711 TI - "Vitamin CE," a novel prodrug form of vitamin E. AB - Reaction of 5a-bromo-alpha-tocopherol with ascorbic acid produces 5a-tocopheryl ascorbate which is designated "vitamin CE." This novel tocopherol derivative represents an interesting prodrug form of alpha-tocopherol (vitamin E) that is stable under acidic conditions, but regenerates finely dispersed vitamin E in basic media. The reaction mechanism of the base-induced decomposition of vitamin CE involves elimination of ascorbate and production of an ortho-quinone methide intermediate that oxidizes ascorbate, and is reduced to vitamin E. Kinetic experiments showed reaction to proceed in the pH range of 8 to 11 under physiological conditions. Tissue culture measurements demonstrated that vitamin E generated from the novel derivative is absorbed at much higher rates than conventional preparations and can even be absorbed under simulated conditions of malabsorption where there is no uptake of conventional vitamin E medications. PMID- 9214712 TI - Enantioselective fluorination of organic molecules. I. Synthetic studies of the agents for electrophilic, enantioselective fluorination of carbanions. AB - In order to develop novel methods for electrophilic and enantioselective fluorination of active methine compounds, preliminary experiments were carried out. The N-tosyl derivative 5 obtained from D-phenylglycine was fluorinated with FClO3 or diluted F2 gas to give the N-fluoro-N-tosyl derivative 6. N-tosyl- or N mesyl-(S)-alpha-phenethylamine 7 or 8 was subjected to FClO3 fluorination to produce the corresponding N-fluoro derivative, 10 or 11, respectively. Enantioselective fluorination of some methine compounds was attempted employing the above N-fluoro agents. Best result was obtained when 2-benzyl-1 tetralone/KHMDS was treated with 10 to produce the fluorinated tetralone 17 in 53% yield with enantiomeric excess (ee) of 48%. PMID- 9214713 TI - Aromatic hydrocarbon quinone-mediated reactive oxygen species production on hepatic microsomes of the flounder (Platichthys flesus L.). AB - The NAD(P)H-dependent redox cycling of a range of eight 1 ring to 5 ring aromatic hydrocarbon (AH) quinones by hepatic microsomes of flounder (Platichthys flesus) was studied in terms of oxygen consumption (Clark electrode) and reactive oxygen species (ROS) production (detection of hydroxyl radical by iron/EDTA-mediated oxidation of 2-keto-4-methiolbutyric acid). Stimulated oxygen consumption was detectable for only five AH-quinones (duroquinone, 1,2- and 1,4-naphthoquinones, menadione, 9,10-phenanthrenequinone), whereas stimulated ROS production was seen, or is known, for all eight (others plus 1,4-benzoquinone, anthraquinone, benzo[a]pyrene-3,6-dione), indicating that the former measurement is a more sensitive assay of redox cycling. Both processes showed Michaelis-Menten kinetics with respect to AH-quinone concentrations, with values for Vmax and apparent Km being, respectively, 146- to 9895-fold and 3- to 344-fold higher for stimulated oxygen consumption than ROS production. Marked correlation in values for both Vmax and apparent Km was seen between stimulated oxygen consumption and ROS production for 1,2-naphthoquinone, 1,4-naphthoquinone and 9,10 phenanthrenequinone, indicative of redox cycling and the univalent reduction of O2 to superoxide anion radical. Rates of stimulated oxygen consumption and ROS production were up to 10-fold higher for NADH- than for NADPH-dependent reactions and were highest for the naphthoquinones and 9,10-phenanthrenequinone. Comparison of the results for different AH-quinones indicates that enzyme substrate specificity is an important factor in determining redox cycling potential. Under the assay conditions used (0.1-2.0 mM AH-quinone), mutagenicity of the AH-quinone mediated processes could not be demonstrated using the Salmonella typhimurium umu assay. Overall, the results indicate a widespread potential for AH-quinone stimulated ROS production. PMID- 9214714 TI - Seasonal variation in the antioxidant defense system of the brain of the ground squirrel (Citellus citellus) and response to low temperature compared with rat. AB - Seasonal variation in the activity of antioxidant enzymes (superoxide dismutase (EC 1.15.1.1.; SOD), catalase (EC 1.11.1.6; CAT), glutathione peroxidase (EC 1.11.1.9; GSH-Px), glutathione reductase (EC 1.6.4.2; GR), glutathione-S transferase (EC 2.5.1.18; GST) and low-molecular-weight antioxidants: ascorbic acid (AsA), vitamin E (VIT E) and glutathione (CSH+GSSG) were examined in the brain of the ground squirrels (Citellus citellus) maintained at 30 degrees C during the whole year. The highest activity (per mg protein) of antioxidant defense (AD) enzymes was found in the spring and was much lower in the summer. A further decrease in activity of CAT, GSH-Px and GST was observed in the winter. The highest levels of AsA and glutathione were recorded in winter in comparison with spring and summer. AD system in the brain of the ground squirrel and rates (maintained at thermoneutrality) exposed to low temperature (4 degrees C) for 3, 6 or 24 hr during the summer was studied as well. Summer was chosen as a period of stable euthermia for ground squirrels and in thermoregulation similar to rats. Consumption of free fatty acid and glucose during the acute exposure to low temperature was found to be species specific. In the ground squirrel, an increase in the specific activities of SOD, after 3, 6 and 24 hr, CAT after 3 and 6 hr and GR after 6 hr of exposure to low temperature was detected. When activities were expressed in U/g wet mass, an increase of SOD after 3, 6 and 24 hr (P < 0.02, P < 0.02, P < 0.005) and CAT and GSH-Px 3 hr (P < 0.01) upon exposure to low temperature was observed. In the rats, no changes in the specific activities of these enzymes after exposure to low temperature were recorded and only an increase in GST activity (U/g wet mass) after 6 hr exposure was registered. Low molecular-weight AD components in both animal species were unchanged upon short term exposure to low temperature. The species-specific differences in brain AD between the rats and the ground squirrels after short exposure to low temperature may be ascribed to seasonal changes of the brain activity in the latter. PMID- 9214715 TI - Antioxidant enzymatic activity in embryos and placenta of rats chronically exposed to hypoxia and hyperoxia. AB - Embryos exhibit lower enzymatic antioxidant activity (EAOA) than adults, in accordance with the low in utero oxygen concentration. We asked whether external oxygen stress can modulate embryonic EAOA and what the placenta role is a mediator between embryos and external milieu. Pregnant rats were exposed to hyperoxia (90% O2) or hypoxia (10% O2) during 8 days in the second or third trimester. Activities of catalase (CAT), superoxide dismutase and glutathione peroxidase (GPx) were measured in the term embryonic brain, lung and heart and liver; 2-week-old whole embryonic sac and placenta. In term "hyperoxic" embryos, only CAT increased by 30% in heart and lungs and liver. In the placenta, GPx increased by 31%. In term "hypoxic" embryos, only CAT activity decreased by 64%, 25% and 29% in brain, liver and placenta. In 2-week-old "hyperoxic" embryos CAT activity increased by 85% and GPx by 45% in the embryonic sac. In the placenta, GPx increased by 55%. The limited embryonic EAOA response is possibly due to maternal physiological buffering of oxygen supply. Placental EAOA is similar to other embryonic organs. It may protect the placenta proper, thus ensuring normal embryonic development. PMID- 9214716 TI - Influence of maternal nicotine exposure on neonatal rat lung structure: protective effect of ascorbic acid. AB - The aims of this study were (1) to determine and quantify the adverse effects of maternal nicotine exposure during pregnancy and lactation on neonatal rat lung development, and (2) to establish whether ascorbic acid will protect the neonatal rat lung against the adverse effects of maternal nicotine exposure. Pregnant rats received nicotine (1 mg/kg body mass/day) subcutaneously during gestation and lactation. A second group received nicotine and ascorbic acid (1 mg/kg body mass/day). The control animals received saline subcutaneously. The results illustrate that maternal nicotine exposure results in (a) a decreased (P < 0.001) radial alveolar count (RAC), (b) an increase (P < 0.001) in destructive index (DI), (c) an increased (P < 0.001) linear intercept (Lm), (d) an increased (P < 0.001) abnormal alveolar attachment index (AAA) (e) and an increase in septal cellularity. Ascorbic acid does not protect fetal lung development against the adverse effects of maternal nicotine exposure. However, after birth ascorbic acid prevents further deterioration of the DI, AAA and Lm, whereas the RAC and thus the number of alveoli was even higher than in control neonatal rat lung. No further increase in cellularity occurred. The reason for this response to ascorbic acid supplementation is under investigation. PMID- 9214717 TI - Effect of cadmium and selenium on the antioxidant defense system in rat kidneys. AB - To examine effects of exogenous Cd on the kidney antioxidant defense system (AOS) and the possible protective role of Se against Cd toxicity, male Wistar albino rats (2 months old) were exposed during 30 days to oral intake of 200 ppm Cd (as CdCl2), 0.l ppm Se (as Na-selenite) or to the same doses of Cd / Se, simultaneously. Marked accumulation of Cd (23.44 +/- 0.69 micrograms/g w.m.) and marked alterations of AOS, resulting in kidney injury (renal pseudohypertrophy), were found in Cd-treated rats. Activities of total superoxide dismutase (SOC, EC 1.15.1.1), manganese-containing superoxide dismutase (MnSOD) and selenium dependent glutathione peroxidase (Se GSH-Px, EG 1.11.1.9) were significantly reduced, whereas that of glutathione-S-transferase (CST, EC 2.5.1.18) and vitamin E (vit E) concentration were significantly increased in the kidneys of Cd-treated rats. Kidney catalase (CAT, EC 1.11.1.6) activity, ascorbic acid (AsA) and red blood cell glutathione (GSH, GSSG) levels were not markedly influenced by CD uptake. In kidneys of Se treated rats, the activities of total SOD, copper-zinc containing superoxide dismutase (CuZnSOD) and GST were significantly increased Activities of kidney CAT and Se GSH-Px were largely unchanged, whereas significant increases of the kidney AsA and vit E concentrations occurred. In Cd + Se-cotreated rats, the kidney activities of MnSOD, CAT and Se GSH-Px, as well as vit E concentration, were the same as in controls, whereas CuZnSOD and GST activities and concentration of AsA exceeded normal values. These data indicate that Se only partially improves the AOS that is insufficient to prevent Cd induced nephrotoxicity. PMID- 9214718 TI - Activity of antioxidant defense enzymes and glutathione content in some tissues of the Belgrade (b/b) laboratory rat. AB - The activity of antioxidant defense (AD) enzymes--superoxide dismutase (SOD, EC 1.15.1.1.), catalase (CAT, EC 1.11.1.6.), glutathione peroxidase (GSH-Px, EC 1.11.1.9.), glutathione-S-transferase (GST, EC 2.5.1.18), glutathione reductase (GR, EC 1.6.4.2) and glutathione (GSH) content of the anemic Belgrade (b/b) laboratory rats--were measured and analyzed in liver, spleen, lung, heart, brain and testes in comparison with nonanemic controls. The activities of hepatic Mn SOD, CAT, GSH-Px and GST (P < 0.02, P < 0.01 and P < 0.005) were decreased in anemic, comparing with nonanemic animals, whereas the spleen CuZn SOD, Mn SOD, CAT and GSH-Px (P < 0.005, P < 0.02, P < 0.005 and P < 0.01) activities were increased. In the lung of anemic rats, Mn SOD, GSH-Px and GR (P < 0.005, P < 0.01, P < 0.05) activities were higher, whereas GST (P < 0.01) activity was lower in relation to nonanemic ones. In anemic rats, heart Mn SOD (P < 0.05) activity was increased, brain GSH-Px (P < 0.005) activity was lower, whereas GR (P < 0.02) activity was higher compared with nonanemic controls. CuZn SOD (P < 0.05) activity in the testes was elevated and GSH-Px (P < 0.05) reduced in anemic animals. GSH content was decreased in the liver (P < 0.01), lung and brain (P < 0.005) and increased in the spleen (P < 0.02) of anemic rats in relation to the controls. Our data suggest phenotype specific differences in the AD system of the Belgrade (b/b) rat tissues in comparison with nonanemic controls. PMID- 9214719 TI - Activities of antioxidant enzymes and monoamine oxidase-A in the rat interscapular brown adipose tissue: effects of insulin and 6-hydroxydopamine. AB - The effects of different doses of insulin (INS) (0.4 or 4.0 IU/kg body mass, i.p., for 3 hr) and 6-hydroxydopamine (6-HDA) (100 mg/kg., i.p.) on the activities of antioxidant enzymes--copper-zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD), catalase (CAT) and catecholamine degrading enzyme monoamine oxidase (MAO-A)--in the rat interscapular brown adipose tissue (IBAT) were studied. In vivo 6-HDA administration, which induces the destruction of sympathetic nerves, markedly reduced IBAT CuZnSOD activity but did not change MnSOD and CAT activities. However, the low dose of INS, which did not induce hypoglycemia, significantly increased the activity of both IBAT mitochondrial enzymes (MnSOD and MAO-A) of control rats. This INS effect on MnSOD was abolished by 6-HDA. On the contrary, CuZnSOD activity was markedly reduced under the influence of INS in both control and 6-HDA-treated rats, whereas for the maintenance of the control level of this enzyme activity, the intact sympathetic nervous system (SNS) is necessary. INS, independent of the dose applied, did not affect CAT activity in control rats, whereas only low INS dose increased the activity of this enzyme in 6-HDA-treated rats. The results indicate that the stimulatory effect of INS on the IBAT mitochondrial enzymes studied is dose dependent and in the case of MnSOD is mediated by SNS. However, the depression in the activity of CuZnSOD is independent of the above-mentioned factors. PMID- 9214720 TI - Chronic effect of insulin on monoamine oxidase and antioxidant enzyme activities in the rat brainstem. AB - It was shown that hydrogen peroxide (H2O2) is a possible intracellular second messenger in specific insulin action. Because its concentration in the cell depends on the activity of both antioxidant enzymes and monoamine oxidase (MAO), we studied the influence of different insulin doses (0.4 and 4.0 IU/kg body mass, i.p., daily injected over 3 days) on the activity of MAO, types A and B, copper zinc superoxide dismutase (CuZnSOD), manganese superoxide dismutase (MnSOD), and catalase in the rat brainstem. Chronic insulin treatment significantly increased Vmax of MAO-A and B activities (P < 0.05, P < 0.025, respectively) independent of the dose applied. CuZnSOD activity was also increased (P < 0.025), but only when higher dose of hormone was injected. However, insulin had the opposite effect on MnSOD and catalase causing a decrease in their activities (P < 0.005). The observed changes in the activities of the enzymes studied are possible compensations that potentially maintain an optimal H2O2 level in the brainstem, which might be important for insulin action. PMID- 9214722 TI - Bibliography of Comparative Biochemistry and Physiology C generated from the Current Awareness in Biological Sciences database. PMID- 9214721 TI - Glucuronidation of thyroxine and 3,5,3'-triiodothyronine by hepatic microsomes in rainbow trout, Oncorhynchus mykiss. AB - We studied the glucuronidation of thyroxine (T4) and 3,5,3'-triiodothyronine (T3) in the hepatic microsomal fraction of rainbow trout, Oncorhynchus mykiss. Uridine diphosphoglucuronosyl transferase (UDPGT) activity toward T4 depended on both protein concentration (linear up to about 0.75 mg/ml) and time (linear up to 60 min). The optimal pH for glucuronidation was 6.8-7.8 units for T4 and > or = 8.5 units for T3. At a common pH of 7.8, the apparent Km and Vmax values were, respectively, 6.0 muM and 42 nmol/mg protein/hr for T4, and 1.1 muM and 4.3 nmol/mg protein/hr for T3. At concentrations of 100 muM, T4 inhibited T3 glucuronidation, but T3 did not inhibit T4-glucuronidation. T4-glucuronidation was inhibited by 3,3', 5'-triiodothyronine (rT3) and tetraiodothyroacetic acid (Tetrac); T3-glucuronidation was inhibited by rT3, T4, Tetrac, and triiodothyroacetic acid. Therefore, analogues with two outer-ring iodines were the most effective inhibitors, showing that outer-ring iodine substitution influences UDPGT substrate specificity. Following a 15-min pre-incubation at 24 degrees C, UDPGT thermal stability was higher for T4 than T3. Polyoxyethylene 10 cetyl ether (Brij 56) maximally stimulated UDPGT activity for both T4 and T3 about two-fold at 0.0125% (w/v) detergent, suggesting that the UDPGTs are transmembrane proteins with the active site facing the lumen of the microsomal vesicles. Clofibrate did not affect either T4- or T3-UDPGT activity. On a per mg microsomal protein basis, T4-glucuronidation in the rat liver exceeded that in the trout 3-fold. We conclude that (a) trout liver microsomes have more than one form of UDPGT for the glucuronidation of T4 and T3 and (b) these trout UDPGTs share several properties of with those of the rat. PMID- 9214723 TI - Effects of some saturated n-alkanes on the survival of Escherichia coli resting cells in sea water. AB - The influence of some linear alkanes on the survival of Escherichia coli natural sea water was investigated. Alkanes with fifteen or more carbon atoms induced a large decrease in viability of E. coli cells in natural sea water. In this case and for concentrations higher than 100 ppm, the loss of viability followed an exponential relationship with the carbon chain length. In the presence of 500 mg l-1 heptane, the survival was 1.6 times higher than that of controls. The progressive disappearance of heptane from the survival medium with a low and temporary accumulation by cells, suggests that this alkane may have been responsible for the increase of cell viability. PMID- 9214724 TI - Issues in the genetic assessment of predispositions for familial breast and ovarian cancer. AB - A rapid development in technology has enabled predictive testing for cancer suceptibility genes, such as BRCA1 and BRCA2. Already biotechnology companies and a number of university-based researchers are establishing service laboratories for the analysis of cancer predispositions. It is critical that high standards be established and maintained when conducting DNA testing, which should be performed in a research setting with proper Institutional Review Board approval. A cancer risk assessment programme should include the involvement of various experts in genetics, oncology, psychiatry and counselling. The legal, ethical and social issues involved in screening and testing for cancer predisposition genes are complex. Genetic counselling of subjects undergoing testing is needed to inform them of the potential risks as well as the potential benefits associated with presymptomatic testing. PMID- 9214726 TI - Immunoreactivities of m-calpain, calpastatin, nitric oxide synthase, myelin basic protein and dynamin II in baker's yeast, wheat germ and lobster tail muscle. AB - Vertebrate m-calpain, calpastatin, constitutive nitric oxide synthase, myelin basic protein, and dynamin I are substrates of protein kinase C (PKC). The presence/absence of similar/related protein in nonvertebrate was investigated by immunological methods, including (1) affinity chromatography on agarose-secondary antibodies and agarose IgG for removal of nonspecific immunoreactivities from crude extracts; (2) omitting beta-mercaptoethanol treatment and boiling prior to SDS-PAGE to increase the immunoreactivity; (3) immunoreactivity comparisons of nonspecific IgG as controls with specific anti-(vertebrate PKC-substrates/related proteins) in Western blots. It was found that (a) m-calpain and dynamin I were absent in baker's yeast, wheat germ and lobster tail muscle, (b) m-calpain, nitric oxide synthase, myelin basic protein and dynamin II were present in all three samples, and (c) calpastatin was present in baker's yeast and lobster tail muscle. The presence and absence of these proteins suggest evolutionary conservation and divergence, respectively, of these PKC substrates. PMID- 9214727 TI - Cardenolides from the methanolic extract of Nerium oleander leaves possessing central nervous system depressant activity in mice. AB - Two new cardenolides, 3 beta-O-(D-2-O-methyldigitalosyl)-14 beta-hydroxy-5 beta carda-16,20(22)-dienolide (1) and 3 beta-hydroxy-8,14-epoxy-5 beta-carda 16,20(22)-dienolide (2), and two known cardenolides, 3 beta-O-(D-digitalosyl)-14 beta-hydroxy-16 beta-acetoxy-5 beta-card-20(22)-enolide (3) and 3 beta-O-(D digitalosyl)-14 beta-hydroxy-5 beta-card-20(22)-enolide (4), have been isolated from the leaves of Nerium oleander following a bioactivity-directed isolation of the MeOH extract, which showed central nervous system (CNS) depressant activity in mice at a dosage of 50 mg/kg i.p. Their structures were established on the basis of chemical and spectral data. Compounds 1, 3, and 4 were found to exhibit sedation in mice at a dosage of 25 mg/kg, although 2 had no effect on the CNS of mice at a dosage of up to 50 mg/kg. PMID- 9214728 TI - Using scalarane sesterterpenes to examine a sponge taxonomic anomaly. AB - A parallel study was conducted on two Indo-Pacific foliose sponges. The first specimen contains 3-hydroxy-20,22-dimethyl-20-deoxoscalarin (2), while the second contains 3-oxo-20,22-dimethyl-20-dioxoscalarin 8 (3). The physical properties as well as X-ray results confirming the structure and stereochemical features of these compounds are presented first. The difficulty we encountered in the taxonomic identification of these species is also discussed. One of our specimens is identical to material considered by different taxonomists as either Phyllospongia vermicularis or Dysidea vermicularis. The other is identified as Carteriospongia sp. We outline that the parallel chemistry of these two specimens suggests that they are closely related taxonomically. PMID- 9214729 TI - Novel mono-tetrahydrofuran ring acetogenins, from the bark of Annona squamosa, showing cytotoxic selectivities for the human pancreatic carcinoma cell line, PACA-2. AB - The bark of Annona squamosa yielded three new mono-tetrahydrofuran (THF) ring acetogenins, each bearing two flanking hydroxyls and a carbonyl group at the C-9 position. These compounds were isolated using the brine shrimp lethality assay as a guide for the bioactivity-directed fractionation. (2,4-cis and trans)-Mosinone A (1) is a mixture of ketolactone compounds bearing a threo/trans/threo ring relationship and s double bond two methylene units away from the flanking hydroxyl. The other two new acetogenins differ in their stereochemistries around the THF ring; mosin B (2) has a threo/trans/erythro configuration across the ring, and mosin C (3) possesses a threo/cis/threo relative stereochemistry. Also found was annoreticuin-9-one (4), a known acetogenin that bears a threo/trans/threo ring configuration and a C-9 carbonyl and is new to this species. The structures were elucidated based on spectroscopic and chemical methods. Compounds 1-4 all showed selective cytotoxic activity against the human pancreatic tumor cell line, PACA-2, with potency 10-100 times that of Adriamycin. PMID- 9214730 TI - Effects of flavonoids isolated from scutellariae radix on fibrinolytic system induced by trypsin in human umbilical vein endothelial cells. AB - Studies on the effects of flavonoids isolated from the roots of Scutellaria baicalensis on the fibrinolytic system induced by trypsin in cultured human umbilical vein endothelial cells (HUVECs) showed that baicalein (1) strongly inhibited the reduction of t-PA production and the elevation of PAI-1 production induced by trypsin. The IC50 for PAI-1 production was 3.7 microM. In addition, wogonin (3), oroxylin A (5), skullcapflavone II (6), and 2',5,5',7-tetrahydroxy 6',8-dimethoxyflavone (7) inhibited the elevation of PAI-1 induced by trypsin, though less strongly; their IC50 were 105, 61, 110, and 88 microM, respectively. These findings suggest that baicalein prevents the thrombotic tendency induced by trypsin. PMID- 9214731 TI - Hypoglycemic activity of some triterpenoid glycosides. AB - Four triterpenoid glycosides isolated from the rhizomes of Polygala senega var. latifolia, senegins II-IV (1-3) and desmethoxysenegin II (4), were tested for hypoglycemic activity in normal and KK-Ay mice. Compounds 1 and 2 reduced the blood glucose of normal mice 4 h after intraperitoneal administration and also significantly lowered the glucose level of KK-Ay mice under similar conditions. Compounds 3 and 4, as well as senegose A (5), an oligosaccharide ester, were inactive when tested against normal mice. PMID- 9214732 TI - Cytotoxic and antiplatelet aggregation principles from Aglaia elliptifolia. AB - Two related 1H-2,3,3a,8b-tetrahydrocyclopenta[b]benzofurans, aglafolin (1a) and rocaglamide (2), isolated from the stems of Aglaia elliptifolia, showed significant cytotoxicity in six cancer cell lines. Aglafolin (1a) was also found to completely block platelet aggregation caused by arachidonic acid and platelet activating factor at 100 microM and 2 ng/mL, respectively. PMID- 9214733 TI - Antihyperglycemic activity of phenolics from Pterocarpus marsupium. AB - Glucose levels in rats with hyperglycemia induced by streptozotocin were determined after i.p. administration of marsupsin (1), pterosupin (2), and pterostilbene (3), three important phenolic constituents of the heartwood of Pterocarpus marsupium. Marsupsin and pterostilbene significantly lowered the blood glucose level of hyperglycemic rats, and the effect was comparable to that of 1,1-dimethylbiguanide (metformin). PMID- 9214734 TI - Psammaplysin F, a new bromotyrosine derivative from a sponge, Aplysinella sp. AB - A new member of the psammaplysin family, psammaplysin F (6) has been isolated from an undescribed species of Aplysinella sponge, along with the four known psammaplysins A-C (1-3) and E (5). The structure of psammaplysin F was determined by spectral analysis. PMID- 9214735 TI - Sustained harvest of camptothecin from the leaves of Camptotheca acuminata. AB - Over a 12-week period, new growth was collected at different intervals from Camptotheca acuminata trees to determine whether a leaf harvest strategy would be an efficient means for the production of the alkaloid camptothecin. Because camptothecin accumulates in young leaves and because the harvesting of young tissue stimulates axillary bud outgrowth, this strategy increased the harvestable amount of camptothecin from trees in a nondestructive manner. PMID- 9214736 TI - Antimicrobial and cytotoxic phenolic glycoside esters from the New Zealand tree Toronia toru. AB - Two new compounds, 4-hydroxyphenyl 6-O-(4-hydroxy-2-methylenebutanoyl)-beta-D glycopyranoside (1) and 4-hydroxyphenyl 6-O-[(3R)-3,4-dihydroxy-2 methylenebutanoyl]-beta-D-glucopyranosid e (2), have been isolated from foliage of Toronia toru. Compound 2 was the main antimicrobial component (7 mg/g of dry foliage) of the crude extract and also showed significant cytotoxic activity against P-388 leukemia cells. PMID- 9214737 TI - Anserinones A and B: new antifungal and antibacterial benzoquinones from the coprophilous fungus Podospora anserina. AB - Two new benzoquinones with antifungal, antibacterial, and cytotoxic activities have been isolated from liquid cultures of the coprophilous fungus Podospora anserina. The structures of anserinones A (1) and B (2) were assigned on the basis of MS and NMR results, and the absolute stereochemistry of 2 was deduced by analysis of 1H-NMR data for its (R)- and (S)-2-phenylbutyryl ester derivatives. PMID- 9214738 TI - Two new lignans with activity against influenza virus from the medicinal plant Rhinacanthus nasutus. AB - Two new lignans, rhinacanthin E (1) and rhinacanthin F (2), were isolated from the aerial parts of the plant Rhinacanthus nasutus. Their structures were established by detailed spectroscopic analysis. These compounds show significant antiviral activity against influenza virus type A. PMID- 9214739 TI - Adenosine-1 active ligands: cirsimarin, a flavone glycoside from Microtea debilis. AB - Several plants collected through different approaches were screened on distinct receptors using ligand-binding studies as bioassay. Extracts of Microtea debilis showed high activity on adenosine A1 receptors. Bioassay-guided fractionation using ligand-binding studies resulted in the isolation of an adenosine A1 active ligand, cirsimarin (cirsimaritin 4'-O-glucoside). GTP did not influence the radioligand inhibition curve of cirsimarin, indicating that this compound is acting as an antagonist at the adenosine-A1 receptors. The use of this plant against "proteinuria" in traditional medicine in Suriname (South America) may be explained by the adenosine A1 antagonistic action of cirsimarin. A series of flavonoids was tested in the same assay, but they were less active. No structure activity relationship could be observed. PMID- 9214740 TI - Bioactive alkaloids from Illigera luzonensis. AB - Using antiplatelet aggregation as a guide to fractionation, seven aporphines, actinodaphnine (1), N-methylactinodaphnine (2), launobine (3), dicentrine (4), O methylbulbocapnine (5), hernovine (7), and bulbocapnine (9), and two oxoaporphines, dicentrinone (6) and liriodenine (8), were isolated from the stems of Illigera luzonensis. Among them, compounds 2, 4, 5, 8, and 9 were isolated for the first time from this species. Moreover, compounds 1-5, and 8 showed significant antiplatelet aggregation and compounds 1 and 6 exhibited significant vasorelaxant activities, respectively. PMID- 9214741 TI - Activity of Parthenolide at 5HT2A receptors. AB - Parthenolide displaces [3H]ketanserin from 5HT2A receptors from rat and rabbit brain and cloned 5HT2A receptors. Ki's are in the 100-250 microM range. These results suggest that parthenolide may be a low-affinity antagonist at 5HT receptors; it is unlikely that the entire mechanism of action can be explained by its modest 5HT2A receptor affinity. PMID- 9214742 TI - Selection on the protein-coding genes of the TBE1 family of transposable elements in the ciliates Oxytricha fallax and O. trifallax. AB - TBE1s are "cut-and-paste" transposable elements found in high copy number in the germline genomes of the ciliates Oxytricha fallax and O. trifallax. TBE1 "family" sequence (sequence of mixed polymerase chain reaction products generated using primers that match roughly half the TBE1s in host whole-cell DNA) was obtained from both host species. Although family sequence autoradiograms represent thousands of different elements, they are as legible as those representing corresponding sequences of a single TBE1, implying that ideal polymorphisms are rare within the genes examined. Nucleotide polymorphisms among TBE1s (indicated by ambiguities in family sequence) are far more common at third than at first or second positions of codons of genes, implying that selection has conserved the amino acid sequences of these genes in the majority of TBE1s. Portions of the transposase gene and another TBE1 gene have been sequenced from 10 individual TBE1s. None of these portions is interrupted by stop codons or frameshifts, and, for both genes, pairwise comparisons of these sequences show that nonsynonymous differences are significantly less common than synonymous differences, again implicating conservative selection Phylogenetic analysis shows that multiple divergent lineages of TBE1s have evolved under this selection within O. fallax. All these results are unexpected for cut-and-paste transposons in eukaryotic hosts: since transposase encoded by intact elements presumably acts in trans, it can duplicate mutant copies (those that do not encode functional transposase) found in the same genome, and thus no selection is expected to maintain the transposase gene. The selection demonstrated here could act at transposition (if functional TBE1s are preferentially transposed) or at the level of the host (if the host's fitness depends on functional TBE1 genes). TBE1-encoded proteins might be responsible for the precise excision of TBE1s that occurs during development of the host somatic nucleus; selection on hosts for uninterrupted somatic genes would then translate into selection for TBE1 protein-coding competence. We suggest a method for distinguishing between these two classes of explanations by finding and analyzing divergent alleles of ancestral transposable element insertions. PMID- 9214743 TI - Comparative nuclear and mitochondrial genome diversity in humans and chimpanzees. AB - Restriction mapping and sequencing have shown that humans have substantially lower levels of mitochondrial genome diversity (d) than chimpanzees. In contrast, humans have substantially higher levels of heterozygosity (H) at protein-coding loci, suggesting a higher level of diversity in the nuclear genome. To investigate the discrepancy further, we sequenced a segment of the mitochondrial genome control region (CR) from 49 chimpanzees. The majority of these were from the Pan troglodytes versus subspecies, which was underrepresented in previous studies. We also estimated the average heterozygosity at 60 short tandem repeat (STR) loci in both species. For a total sample of 115 chimpanzees, d = 0.075 +/0 0.037, compared to 0.020 +/- 0.011 for a sample of 1,554 humans. The heterozygosity of human STR loci is significantly higher than that of chimpanzees. Thus, the higher level of nuclear genome diversity relative to mitochondrial genome diversity in humans is not restricted to protein-coding loci. It seems that humans, not chimpanzees, have an unusual d/H ratio, since the ratio in chimpanzees is similar to that in other catarrhines. This discrepancy in the relative levels of nuclear and mitochondrial genome diversity in the two species cannot be explained by differences in mutation rate. However, it may result from a combination of factors such as a difference in the extent of sex ratio disparity, the greater effect of population subdivision on mitochondrial than on nuclear genome diversity, a difference in the relative levels of male and female migration among subpopulations, diversifying selection acting to increase variation in the nuclear genome, and/or directional selection acting to reduce variation in the mitochondrial genome. PMID- 9214744 TI - Bayesian phylogenetic inference using DNA sequences: a Markov Chain Monte Carlo Method. AB - An improved Bayesian method is presented for estimating phylogenetic trees using DNA sequence data. The birth-death process with species sampling is used to specify the prior distribution of phylogenies and ancestral speciation times, and the posterior probabilities of phylogenies are used to estimate the maximum posterior probability (MAP) tree. Monte Carlo integration is used to integrate over the ancestral speciation times for particular trees. A Markov Chain Monte Carlo method is used to generate the set of trees with the highest posterior probabilities. Methods are described for an empirical Bayesian analysis, in which estimates of the speciation and extinction rates are used in calculating the posterior probabilities, and a hierarchical Bayesian analysis, in which these parameters are removed from the model by an additional integration. The Markov Chain Monte Carlo method avoids the requirement of our earlier method for calculating MAP trees to sum over all possible topologies (which limited the number of taxa in an analysis to about five). The methods are applied to analyze DNA sequences for nine species of primates, and the MAP tree, which is identical to a maximum-likelihood estimate of topology, has a probability of approximately 95%. PMID- 9214745 TI - Nonrandom location of IS1 elements in the genomes of natural isolates of Escherichia coli. AB - We have studied the spatial distribution of IS1 elements in the genomes of natural isolates comprising the ECOR reference collection of Escherichia coli. We find evidence for nonrandomness at three levels. Many pairs of IS1 elements are in much closer proximity (< 10 kb) than can be accounted for by chance. IS1 elements in close proximity were identified by long-range PCR amplification of the genomic sequence between them. Each amplified region was sequenced and its map location determined by database screening of DNA hybridization. Among the ECOR strains with at least two IS1 elements, 54% had one or more pairs of elements separated by < 10 kb. We propose that this type of clustering is a result of "local hopping," in which we assume that a significant proportion of tranposition events leads to the insertion of a daughter IS element in the vicinity of the parental element. A second level of nonrandomness is found in strains with a modest number of IS1 elements that are mapped through the use of inverse PCR to amplify flanking genomic sequences: in these strains, the insertion sites tend to be clustered over a smaller region of chromosome than would be expected by chance. A third level of nonrandomness is observed in the composite distribution of IS elements across strains: among 20 mapped IS1 elements, none were found in the region of 48-77 minutes, a significant gap. One region of the E. coli chromosome, at 98 min, had a cluster of IS1 elements in seven ECOR strains of diverse phylogenetic origin. We deduce from sequence analysis that this pattern of distribution is a result of initial insertion in the most recent common ancestor of these strains and therefore not a hot spot of insertion. Analysis using long-range PCR with primers for IS2 and IS3 also yielded pairs of elements in close proximity, suggesting that these elements may also occasionally transpose by local hopping. PMID- 9214746 TI - Can three incongruence tests predict when data should be combined? AB - Advocates of conditional combination have argued that testing for incongruence between data partitions is an important step in data exploration. Unless the partitions have had distinct histories, as in horizontal gene transfer, incongruence means that one or more data support the wrong phylogeny. This study examines the relationship between incongruence and phylogenetic accuracy using three tests of incongruence. These tests were applied to pairs of mitochondrial DNA data partitions from two well-corroborated vertebrate phylogenies. Of the three tests, the most useful was the incongruence length difference test (ILD, also called the partition homogeneity test). This test distinguished between cases in which combining the data generally improved phylogenetic accuracy (P > 0.01) and cases in which accuracy of the combined data suffered relative to the individual partitions (P < 0.001). In contrast, in several cases, the Templeton and Rodrigo tests detected highly significant incongruence (P < 0.001) even though combining the incongruent partitions actually increased phylogenetic accuracy. All three tests identified cases in which improving the reconstruction model would improve the phylogenetic accuracy of the individual partitions. PMID- 9214748 TI - The mtDNA sequence of the ostrich and the divergence between paleognathous and neognathous birds. AB - The complete mitochondrial DNA (mtDNA) molecule of the ostrich, Struthio camelus, was sequenced. The size of the molecule is 16,591 nucleotides. Since the ostrich represents the paleognathous birds, comparison with the mtDNA of the neognathous chicken, the only avian species reported so far in databases, made it possible to identify common and, probably, general avian mtDNA characteristics. Relative to other vertebrates, the avian NADH6 and tRNA-Glu genes are positioned upstream of the control region rather than the cytochrome b gene. The NADH3 gene of the ostrich is terminated by a stop codon at position 207. Thus, the gene is about 140 nucleotides shorter than in other vertebrates. The sequence for L-strand origin of replication is missing in both birds, and four transfer RNA genes of the two avian mtDNAs deviate from common characteristics of tRNAs of vertebrate mtDNAs by having an adenine (and not a thymidine) at position 8. Due to the absence of suitable fossils, most paleontological datings of avian divergences are conjectural. Molecular dating of the divergence between the ostrich and the chicken indicates that these two avian lineages separated 80-90 MYA. Phylogenetic analysis of complete cytochrome b genes of six avian orders showed that Passeriformes represent the earliest divergence among recent birds, contradicting the commonly accepted notion of a basal position of the Palaeognathae among recent birds. PMID- 9214747 TI - Interspecific and intraspecific comparisons of the period locus in the Drosophila willistoni sibling species. AB - The period (per) locus has received much attention in molecular evolution studies because it is one of the best studied "behavioral genes" and because it offers insight into the evolution of repetitive sequences. We studied most of the coding region of per in Drosophila willistoni and confirmed previously observed patterns of conservation and divergence among distantly related species. Five regions are so highly diverged that they cannot be aligned, whereas a region encompassing the PAS domain is very conserved. Structural and nucleotide polymorphism patterns in the willistoni group are not the same as those observed in previously studied species. We sequenced the region homologous to the highly polymorphic threonine glycine repeat of D. melanogaster in multiple strains of D. willistoni, as well as in other members of willistoni group, and found an unusual amount of conservation in this region. However, the next nonconserved region downstream in the sequence is quite variable and polymorphic for the number of repeated glycines. The glycine codon usage is significantly different in this glycine repeat as compared to other parts of the gene. We were able to plot the directionality of change in the glycine repeat region onto a phylogeny and find that the addition of glycines is the general trend with the diversification of the willistoni group. PMID- 9214749 TI - Phylogenetic analyses of mitochondrial DNA suggest a sister group relationship between Xenarthra (Edentata) and Ferungulates. AB - Phylogenetic analysis of 12 protein-coding genes from complete mitochondrial DNA (mtDNA) molecules of various mammals including a xenarthran representative, the armadillo (Dasypus novemcinctus), showed that the order Xenarthra (Edentata) is a sister group to the ferungulates (carnivores, perissodactyls, artiodactyls, cetaceans). Morphological and previous molecular analyses have placed the Xenarthra basal to other extant eutherians. The present findings are in striking contrast with that understanding. The results suggest that Xenarthra and ferungulates separated about 86 MYA. PMID- 9214751 TI - Purification of characterization of soybean allergen Gly m Bd 28K. AB - At least 15 allergenic proteins have been found in soybean using the sera of soybean-sensitive patients with atopic dermatitis [T. Ogawa et al., J. Nutr. Sci. Vitaminol., 35, 555-565 (1991)]. In the present study, a monoclonal antibody (mAb) against Gly m Bd 28K, one of the major allergens of soybean, was prepared, and Gly m Bd 28K was purified from defatted soybean flakes by five purification steps, including immunoaffinity chromatography with the mAb as a ligand. The purified allergen was found to be a glycoprotein with a molecular mass of 26 kDa. During the purification process the allergen was converted to more acidic proteins with the same molecular mass, suggesting that the allergen is unstable. The sugar composition and amino acid sequence of Gly m Bd 28K suggest that the allergen is a new glycoprotein with an Asn-linked sugar moiety. The distribution of the allergen in soybean products was examined by an immunoblotting technique with the mAb. PMID- 9214750 TI - Comparisons of the molecular evolutionary process at rbcL and ndhF in the grass family (Poaceae). AB - We examine rate heterogeneity among evolutionary lineages of the grass family at two plasmid loci, ndhF and rbcL, and we introduce a method to determine whether patterns of rate heterogeneity are correlated between loci. We show both that rates of synonymous evolution are heterogeneous among grass lineages and that are heterogeneity is correlated between loci at synonymous sites. At nonsynonymous sites, the pattern of rate heterogeneity is not correlated between loci, primarily due to an aberrant pattern of rate heterogeneity at nonsynonymous sites of rbcL. We compare patterns of synonymous rate heterogeneity to predictors based on the generation time effect and the speciation rate hypotheses. Although there is some evidence for generation time effects, neither generation time effects nor speciation rates appear to be sufficient to explain patterns of rate heterogeneity in the grass plastid sequences. PMID- 9214752 TI - Heterologous protein production in Acremonium chrysogenum: expression of bacterial cephalosporin C acylase and human thrombomodulin genes. AB - We have developed an efficient expression system for foreign genes in Acremonium chrysogenum. After inserting the foreign gene between the phosphoglycerate kinase (PGK) promoter and a terminator derived from A. chrysogenum, multiple copies of this expression unit are tandemly ligated into cosmids and the resultant cosmids are introduced into A. chrysogenum. We expressed Pseudomonas cephalosporin C acylase and a human thrombomodulin mutant protein containing the fourth, fifth, and sixth epidermal growth factor (EGF)-like structures (E456). The acylase activity in the transformants obtained using our system was several times higher than that in the transformants without the use of the system. The acylase proteins expressed had enzymatic and immunochemical properties identical to those of authentic acylase. The transformants with the expression plasmid for E456 secreted biologically active E456 protein into the culture medium. The amino terminal sequence of the purified E456 was identical to that of recombinant E456 obtained using mammalian cells. PMID- 9214753 TI - Enzymatic production of pyrimidine nucleotides using Corynebacterium ammoniagenes cells and recombinant Escherichia coli cells: enzymatic production of CDP-choline from orotic acid and choline chloride (Part I). AB - Enzymatic production of cytidine diphosphate choline (CDP-choline) using orotic acid and choline chloride as substrates was investigated using a 200-ml beaker as a reaction vessel. When Cornybacterium ammoniagenes KY13505 cells were used as the enzyme source, UMP was accumulated up to 28.6 g/liter (77.6 mM) from orotic acid after 26 h of reaction. In this reaction, UDP and UTP were also accumulated, but CTP, a direct precursor of CDP-choline, was not accumulated sufficiently. Escherichia coli JF646/pMW6 cells, which overproduce CTP synthetase by selfcloning of the pyrG gene, were used together with cells of KY12505 for the enzymatic reaction using orotic acid as a substrate. CTP was produced at 8.95 g/liter (15.1 mM) after 23 h of this reaction. To produce CDP-choline, two additional enzyme activities were needed. E. coli MM294/pUCK3 and MM294/pCC41 cells, which express a choline kinase from Saccharomyces cerevisiae (CKIase; encoded by the CKI gene) and a cholinephosphate cytidylyltransferase from S. cerevisiae (CCTase; encoded by the CCT gene) respectively, were added to this CTP producing reaction system. After 23 h of the reaction using orotic acid and choline chloride as substrates, 7.7 g/liter (15.1 mM) of CDP-choline was accumulated without addition of ATP or phosphoribosylpyrophosphate (PRPP). ATP and PRPP required in the CDP-choline forming reaction system are biosynthesized by those cells using glucose as a substrate. PMID- 9214754 TI - Construction of a plasmid carrying both CTP synthetase and a fused gene formed from cholinephosphate cytidylyltransferase and choline kinase genes and its application to industrial CDP-choline production: enzymatic production of CDP choline from orotic acid (Part II). AB - A new method for enzymatic production of cytidine diphosphate choline (CDP choline) from orotic acid and choline chloride was developed. To establish an industrial manufacturing process, we constructed a plasmid, pCKG55, which simultaneously expressed in Escherichia coli the three following enzymes; CTP synthetase (encoded by the pyrG gene from E. coli), cholinephosphate cytidylyltransferase (encoded by the CCT gene from Saccharomyces cerevisiae), and choline kinase (encoded by the CKI gene from S. cerevisiae). CCT and CKI genes on pCKG55 were designed to be expressed as a single CCT/CKI fused protein. This CCT/CKI fused protein retained both activities and the thermal stability of its cholinephosphate cytidylyltransferase activity was nearly the same as the native CCT enzyme. Corynebacterium ammoniagenes KY13505 and E. coli MM294/pCKG55 were cultured in 5-liter jar fermentor independently. Equal volumes of each broth were mixed in a 2-liter jar fermentor, and then the enzymatic reaction was done using 47 mM orotic acid and 60 mM choline chloride as substrates. After 23 h of the reaction at 32 degrees C, 21.5 mM (11 g/liter) of CDP-choline was accumulated. PMID- 9214755 TI - Purification and characterization of a glucose-tolerant beta-glucosidase from Aspergillus niger CCRC 31494. AB - An extracellular glucose-tolerant beta-glucosidase was purified to homogeneity by alcohol fractionation and preparative isoelectric focusing from Aspergillus niger CCRC 31494. The enzyme was a dimeric protein with a subunit of 49,000, and had its optimum activity at pH 5.0 and 55 degrees C. The enzyme was completely inhibited by 5 mM Ag+. Thiol groups and serine residues were not essential for its activity. Low concentrations of alcohols (10%) except for methanol could activate the enzyme. It was very specific for para-nitrophenyl-beta-D-glucoside (pNPG) and cellobiose. However, the enzyme also had some beta-xylosidase activity, but showed no activity towards alpha-linked glycosidic substrates. The Vmax of 124.4 U/mg and 21.6 U/mg were found for pNPG (Km = 21.7 mM) and para nitrophenyl-beta-D-xyloside (pNPX) (Km = 14.2 mM), respectively. The enzyme was tolerant to glucose inhibition with a Ki of 543 mM, while fructose, galactose, mannose, and xylose were not inhibitory. PMID- 9214756 TI - Long-term culture of primary rat hepatocytes on heparin- or lambda carrageenan containing collagen gels. AB - The interactions of glycosaminoglycans (GAGs) with collagen are thought to be important in cell adhesion and cell differentiation. To investigate whether the interactions of GAG or sulfated polysaccharide with collagen can maintain the functions of cultured primary rat hepatocytes, GAG- or sulfated polysaccharide containing collagen gels were reconstituted in vitro and used for culture of hepatocytes. Among the GAGs and sulfated polysaccharides examined, heparin- and lambda carrageenan-containing collagen gels were found to be able to stimulate and sustain albumin synthesis, while the other GAG- or sulfated polysaccharide containing collagen gels had almost no effect on maintenance of albumin synthesis. In the cultures using collagen gels that contained 400 micrograms/ml heparin or 100 micrograms/ml lambda carrageenan, albumin synthesis by rat hepatocytes was prolonged to about 4 and 5 weeks, respectively, but albumin synthesis was kept up for only one week in the cultures using conventional collagen gels. These results suggest that the interactions of heparin or lambda carrageenan might be of importance for long-term maintenance of hepatocyte functions. PMID- 9214757 TI - Elicitor actions of N-acetylchitooligosaccharides and laminarioligosaccharides for chitinase and L-phenylalanine ammonia-lyase induction in rice suspension culture. AB - When a series of chitin oligosaccharides was added into a rice suspension culture, N-acetylchitohexaose, N-acetylchitopentaose, and N-acetylchitotetraose caused an increase in extracellular chitinase activity, mainly due to induction of a class III chitinase. In the case of N-acetylchitohexaose, a substantial increase in the chitinase activity was observed at a concentration higher than 0.01 micrograms/ml, and a maximum effect was reached at 1 microgram/ml. In contrast, N-acetylchitotriose, N-acetylchitobiose, N-acetyl-D-glucosamine, and chitohexaose (a chitosan oligosaccharide) were not very effective. Chitinase induction was also observed with laminarihexaose (a beta-1,3-glucan oligosaccharide), but about a 10-fold higher concentration, compared with N acetylchitohexaose, was needed to get the maximum effect. beta-1,3-Glucanase activity was found in cells (but not in medium), and the activity was increased by neither N-acetylchitohexaose nor laminarihexaose. When cells were incubated with N-acetylchitohexaose, L-phenylalanine ammonia-lyase (PAL) activity increased promptly. A biphasic profile was obtained when a dose-dependent effect of the elicitor on the PAL induction was examined; the first phase was observed in a range from 0.01 to 1 microgram/ml and the second phase from 3 to 300 micrograms/ml. Laminarihexaose also acted as an elicitor for PAL induction. PMID- 9214758 TI - The sugar specificity of Na+/glucose cotransporter from rat jejunum. AB - A cDNA for a Na+/glucose cotransporter was cloned from rat jejunum cDNA library. This transporter was expressed in Xenopus oocytes by injection of cRNA synthesized from the cDNA, and the transporter ability was electrophysiologically examined. The cotransporter had a very narrow sugar specificity. Only D-glucose, D-galactose, and some of their derivatives elicited significant electrical responses. These results of sugar specificity were compared with those of the H+/hexose cotransporter of Chlorella. Dose-response relationships of several sugars followed a simple Michaelis-Menten type of kinetics. Both Vm and Km were dependent on the sugars. Not only the affinity of sugars to the cotransporter but also the rate of conformational change of the cotransporter loaded with the sugar and Na+, which translocates them from outside to inside, possibly depends on the sugar structure. The rate-limiting step of the transportation may be the conformational change, i.e., isomerization, of the cotransporter that translocates both the sugar and Na+ from outside to inside. PMID- 9214759 TI - Primary structure of 6.5k-arginine/glutamate-rich polypeptide from the seeds of sponge gourd (Luffa cylindrica). AB - The amino acid sequence of 6.5k-arginine/glutamate rich polypeptide (6.5k-AGRP) from the seeds of sponge gourd (Luffa cylindrica) has been determined. The 6.5k AGRP consists of a 47-residue polypeptide chain containing two disulfide bonds, and a molecular mass calculated to be 5695 Da, which fully coincides with a value of [M+H]+ = m/zeta 5693.39 obtained by matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS). The mass spectrometric evidence indicated that 6.5k-AGRP is also present partially truncated at the C-terminus. In our preparations, approximately half of the polypeptide molecules have the C-terminal sequence Arg-Arg-Glu-Val-Asp; the other half lack Val-Asp and end with the glutamic acid, making a total of 45 residues in the polypeptide chain. The two disulfide bonds connect Cys12 to Cys33 and Cys16 to Cys29. Comparison of the amino acid sequence of 6.5k-AGRP with those of the other known proteins included in the PIR protein sequence database showed that it is related to the amino acid sequence of the N-terminal region encoded by the first exon of the cocoa (Theobroma cacao) and cotton seeds vicilin genes, sharing a characteristic two Cys-Xaa-Xaa-Xaa-Cys motif. PMID- 9214760 TI - Actions of pokeweed antiviral protein on virus-infected protoplasts. AB - Pokeweed antiviral protein (PAP) belongs to a group of ribosome-inactivating proteins (RIPs) that inactivate ribosomes by depurinating rRNA at a specific site. To study the mechanism for the antiviral activity of PAP, the actions of PAP on TMV-infected and uninfected tobacco protoplasts were investigated. The addition of 0.33 microM PAP to TMV-inoculated protoplasts caused a complete inhibition of TMV production. The same concentration of PAP was found to inhibit protein synthesis in the virus-infected protoplasts and to kill the cells, but it had no effect on the uninfected protoplasts. The concentration dependence of protein synthesis-inhibition by PAP was related to that of inhibition of viral multiplication. Furthermore, two other RIPs (ricin A-chain and luffin-a), which showed 240 and 430-fold less activity on tobacco ribosomes than PAP in a cell free system, did not inhibit viral multiplication even at a concentration of 3.3 microM. The analysis of RNAs from the virus-infected and PAP-treated protoplasts demonstrated that 25S rRNA was depurinated by PAP in the infected cells. These results suggest that PAP, which is normally unable to penetrate the plasma membrane of uninfected protoplasts, gains entrance to the cytosol of infected cells and prevents viral multiplication by inactivating ribosomes. PMID- 9214761 TI - Changes in proteasome levels in spinach (Spinacia oleracea) seeds during imbibition and germination. AB - Proteasomes are the major cytosolic protease complexes responsible for energy dependent and extra-lysosomal proteolysis. Ubiquitinated proteins are degraded by the 26S proteasome which is composed of a 20S proteasome and two regulatory complexes. Changes in the 20S and 26S proteasome activity levels and protein abundance in spinach seeds during imbibition and germination were examined by using glycerol density gradient centrifugation. The 26S proteasome activity level decreased transiently during imbibition, reached a minimum one day after starting imbibition, and then increased again. During the period of minimal accumulation, the protein being detected with an anti-26S proteasome antibody shifted to a lower sedimentation coefficient fraction. In contrast, the 20S proteasome activity level increased during imbibition, and remained high for two days. The change in the 20S proteasome level corresponded to the change in the 20S proteasome activity level. PMID- 9214762 TI - Changes in messenger RNA of pancreatic enzymes and intestinal cholecystokinin after a 7-day bile-pancreatic juice diversion from the proximal small intestine in rats. AB - We have previously demonstrated the bile-pancreatic juice (BPJ)-independent stimulation of pancreatic enzyme secretion in chronic BPJ-diverted rats. Pancreatic and intestinal adaptation to 7-day BPJ diversion was next examined. Pancreatic enzyme mRNA and cholecystokinin mRNA in the jejunal mucosa were measured in rats with BPJ diverted into the ileum (PBD rats) in comparison with the figures for rats with BPJ returned to the duodenum (normal rats) or laparotomized (Intact) rats under well-nourished conditions. Amylase mRNA in the pancreas was lower and trypsinogen plus chymotrypsinogen mRNA was higher in the PBD rats than in the intact rats. The change in pancreatic mRNA was similar to that in the specific activities of the enzymes after a chronic BPJ diversion. This finding suggests that these pancreatic enzymes were regulated by the mRNA level. The portal concentration of cholecystokinin in the postabsorptive period (exogenously non-stimulated status) was 4-fold higher in the PBD group than in the normal and intact groups. Cholecystokinin mRNA in the jejunal mucosa of PBD rats was somewhat higher than that of intact rats. These results suggest that intestinal cholecystokinin was predominantly increased at the translational or later stage by chronic BPJ diversion. PMID- 9214763 TI - Novel amino acid metabolite produced by Streptomyces sp.: I. Taxonomy, isolation, and structural elucidation. AB - A strain of streptomycete isolated from a soil sample was found to produce a novel amino acid metabolite. The compound was purified from the culture fluid by chromatography, using cation exchange resin, a synthetic adsorbent, and finally by preparative HPLC with a reverse-phase column. The structure of the compound was established as N(delta)-(5-methyl-4-oxo-2-imidazolin-2-yl)-L-ornithine on the basis of an analysis of the spectral data and chemical degradation. This was confirmed by comparing the NMR spectrum of the metabolite with that of the compound synthesized by treating methylglyoxal and N(alpha)-acetyl-L-arginine. The substance did not show any antimicrobial activity against bacteria, fungi and yeasts by the agar plate method, but exhibited a weak preventive effect on cucumber mildew disease in a pot test. PMID- 9214764 TI - Effects of DNA topology on transformation efficiency of Bacillus subtilis ISW1214 by electroporation. AB - We report an investigation of electrotransformation by three different topological isomers, circular supercoiled (sc DNA), circular relaxed (cr DNA), and linearized (In DNA) forms of the plasmids pUB110 (4.5 kbp) and pBDR331T (12.6 kbp), of a Gram-positive bacterium, Bacillus subtilis ISW1214. Treatment of the sc DNA with calf thymus topoisomerase I removed the superhelicity and the DNA assumed the relaxed circular form. Treatment of sc DNA with restriction endonculease linearized the DNA. The transformation with the sc DNA of pUB110 resulted in the maximum efficiency of (2.6 +/- 0.6) x 10(5) transformants per microgram DNA higher than that (2.0 +/- 0.3) x 10(4) transformants per microgram DNA for the cr DNA, using the DNA concentration of 20 micrograms/ml at an electric field strength of 7 kV/cm and a capacitance of 10 microF with a single decayed pulse. The transformation efficiency (TE) for the ln DNA was zero. The variations of TE for different topological forms of DNA reflected their relative stability in the host cells. The molecular efficiency (ME, transformants per molecule) for sc DNA was nearly one order of magnitude greater for the lower molecular size of pUB110 DNA than that for the higher molecular size of pBDR331T DNA. PMID- 9214765 TI - Inactivation of Bacillus spores by the supercritical carbon dioxide micro-bubble method. AB - Bacillus spores were effectively inactivated by the supercritical (SC) CO2 micro bubble method. The micro-bubble SC CO2 treatment of B. cereus, B. subtilis, B. megaterium, B. polymyxa, and B. coagulans at 40 degrees C and 30 MPa for 30 min produced greater reduction (about 3 log cycles of reduction) than a similar treatment without a filter. The SC CO2 treatment of B. polymyxa, B. cereus, and B. subtilis spores at 45 degrees C, 50 degrees C, respectively, and 30 MPa for 60 min resulted in a 6-log cycle reduction of survival. The SC CO2 treatment under the foregoing conditions should offer higher efficiency than that of heat treatment at 100 degrees C for 60 min. In addition, the SC CO2 treatment (30 MPa, 60 degrees C, 30 min) of B. polymyxa and B. cereus spores also produced a 6-log cycle reduction. PMID- 9214766 TI - Apoptosis to HL-60 by humulone. AB - Humulone, a bone resorption inhibitor isolated from hop extract, induced apoptosis in the premyocytic leukemia cell line HL-60 between 1 and 100 micrograms/ml. Our data suggested that there was a correlation between the apoptosis-inducing activity of humulone and its antioxidative activity. PMID- 9214767 TI - Inhibitory effect of alginic acids on hyaluronidase and on histamine release from mast cells. AB - The effects of various types of alginic acid consisting of L-guluronic acids (G) and D-mannuronic acids (M) on hyaluronidase and mast cell degranulation were examined. Alginic acid with an M/G ratio of 1.0 exhibited the strongest inhibition of both activities, the higher molecular weight alginic acids of 150 to 370 kDa being preferable in both cases. Esterification of the carboxyl residue enhanced the latter activity. PMID- 9214768 TI - Structural analysis of disaccharides synthesized by beta-D-glucosidase of Bifidobacterium breve clb and their assimilation by Bifidobacteria. AB - Circulation of a solution of 1 M D-fucose and 1 M D-glucose through a reaction system consisting of serial columns of immobilized recombinant beta-D-glucosidase of Bifidobacterium breve clb and activated charcoal gave two oligosaccharides. Structural analysis identified these oligosaccharides as D-fucosylglucose (6-O beta-D-Fucopyranosyl-D-glucose) and gentiobiose (6-O-beta-D-Glucopyranosyl-D glucose). The D-fucosylglucose obtained was well assimilated by many Bifidobacteria but not by the other intestinal bacteria tested. PMID- 9214769 TI - Cloning, expression, and mutagenesis of trypsin inhibitor ETIb from Erythrina variegata seeds. AB - Erythrina variegata trypsin inhibitors designated ETIa and ETIb belong to the Kunitz family trypsin inhibitor, but ETIa is unique in its ability to inhibit tissue-type plasminogen activator, while ETIb is not. The cDNA clone encoding ETIb was isolated from the seed cDNA library constructed in the lambda phage lambda gt11. The ETIb cDNA insert consists of 765 bp, including an open reading frame of 606 pb from ATG to TGA codons. The deduced amino acid sequence consists of 202 amino acids, having the signal peptides of 22 amino acids in the N terminus and 2 amino acids in C-terminus. The cDNA fragment encoding the mature form of ETIb was introduced into an expression vector, pET-22b, and expressed in Escherichia coli BL21 (ED3) in a functional form. Furthermore, the ETIb mutant bP61R/F62L, in which Pro61 and Phe62 in ETIb were changed to the corresponding amino acid residues Arg and Leu, respectively, as in ETIa, was constructed, and its inhibitory potency toward tPA was assayed. This mutant showed significant tPA inhibitory activity, albeit less than ETIa. The result demonstrates that the Arg61 and Leu62 residues in ETIa are important in inhibiting tPA, and also suggest that beside these two residues, the other amino acid(s) or other structural element may be involved in interaction of ETIa with tPA. PMID- 9214770 TI - Competitive ELISA of bovine lactoferrin with bispecific monoclonal antibodies. AB - A mouse hybrid-hybridoma, HH1-4-3, secreting IgG1 class bispecific antibodies to bovine lactoferrin (bLF) and horseradish peroxidase (HRPO), was established previously. The competitive enzyme-linked immunosorbent assay (ELISA) of bLF using the HH1-4-3 culture supernatant was not sensitive enough to measure bLF concentration in biological fluids. To improve the sensitivity of the competitive ELISA, we fractionated the bispecific antibodies by antigen affinity column chromatography. A column immobilized with bLF adsorbed 60% of the antibodies of the HH1-4-3 supernatant, and the amount of antibodies adsorbed on a column immobilized with HRPO was less than 5%. The competitive ELISA of bLF using the affinity purified bispecific antibodies through an HRPO-immobilized column chromatography showed a good standard curve at bLF concentrations of 10 ng/ml to 100 micrograms/ml. PMID- 9214771 TI - A bacteriocin of strain Pediococcus sp. ISK-1 isolated from Nukadoko, bed of fermented rice bran. AB - Pediococcus sp. ISK-1 isolated in our laboratory from well-aged Nukadoko, produces a bacteriocin which has a unique antimicrobial spectrum among pediocins. The bacteriocin was stable at acidic pH, and more than 60% of antimicrobial activity still remained even after being autoclaved at 121 degrees C for 20 min in the pH range of 3 to 8. This is the first report dealing with a bacteriocin produced by lactic acid bacteria isolated from Nukadoko. PMID- 9214772 TI - Determination of endogenous peptides with in vitro ACE inhibitory activity in normotensive human plasma by the fluorometric HPLC method. AB - An in vitro degradation test of angiotensin (ANG) II or III in normotensive supine human plasma from 9 healthy male subjects confirmed the production of smaller ANG metabolites with angiotensin I-converting enzyme inhibitory activity. These metabolites were identified as ANG (3-8), ANG (5-8), and ANG (3-4), whose respective peptide concentrations were determined by our proposed naphthalene-2,3 dialdehyde (NDA)-HPLC method to be 64 +/- 9, 39 +/- 5, 176 +/- 22, and 197 +/- 35 fmol/ml of plasma. PMID- 9214773 TI - Purification and properties of a new enzyme, D-carnitine dehydrogenase, from Agrobacterium sp. 525a. AB - A new enzyme, D-carnitine dehydrogenase from Agrobacterium sp. 525a, was purified by DEAE-Toyopearl, ammonium sulfate fractionation, Sephadex G-75, affinity chromatography, and Mono Q and TSK-gel filtration column chromatography. The enzyme had the molecular mass of 89 kDa and consisted of three identical subunits. The optimum pH for the oxidation reaction was 9.3. The Michaelis constants for D-carnitine and NAD+ were 3.1 and 0.07 mM, respectively. The N terminal 20 amino acids were sequenced. PMID- 9214774 TI - Amino acid sequence of the basic subunit of 13S globulin of buckwheat. AB - A 26 kDa basic subunit of 13S globulin has been purified from grains of common buckwheat (Fagopyrum esculentum Moench). The amino acid composition of the protein closely matches the W.H.O. recommended values for a nutritionally balanced protein. The sequence of 17 N terminal amino acid residues of the protein revealed 73.3 and 66.7% homology with soya bean glycinin and pea legumin, respectively. PMID- 9214775 TI - Oxidation of the phytoalexin maackiain to 6,6-dihydroxy-maackiain by Colletotrichum gloeosporioides. AB - Phytoalexins are low molecular weight antibiotic compounds produced by plants in response to infection by microbes. These antimicrobial compounds are thought to provide resistance to microbial invasion and colonization. (-)Maackiain and its pterocarpan relatives can be oxidized at a number of sites, including at the 6 carbon. A previously unknown metabolite was produced for (-)maackiain by the broad host-range pathogen Colletotrichum gloeosporioides (Glomerella cingulata). This unknown was identified by LC-MS-MS and NMR spectroscopy to be 6,6a-di-OH maackiain (3,6,6a-trihydroxy-8,9,methylenedioxy-pterocarpan). It is produced by isolates that represent all four races and pathotypes of C. gloeosporioides isolated from the tropical forage legume Stylosanthes spp. We present evidence that the primary metabolite (-)6a-OH-maackiain is subsequently hydroxylated at carbon 6, a step resulting in a compound that is increased in polarity and decreased in toxicity relative to the parent compound and (-)6a-OH-maackiain. This further oxidation may be required for efficient excretion or carbon source scavenging. PMID- 9214776 TI - Induction of ABA 8'-hydroxylase by (+)-S-, (-)-R- and 8'-8'-8'-trifluoro-S abscisic acid in suspension cultures of potato and Arabidopsis. AB - Suspension cultures of potato and Arabidopsis were incubated with 50 microM of (+)-ABA and (-)-ABA for 3 hr. These pretreatments were found to increase the rate, by two- to seven-fold, of formation of [2H6] phaseic acid (PA) from [2H6] ABA, applied in a subsequent incubation. Pretreatment with trifluoro-ABA had a higher efficacy, increasing the rate of conversion 15-fold. Suspension cell cultures that had been dehydrated and then rehydrated in the presence of [2H6] ABA displayed a much lower enhancement of PA formation. We conclude that ABA induces its own oxidative catabolism in suspension cultures. PMID- 9214777 TI - Biological activities of some Argyranthemum species. AB - Seven species of the genus Argyranthemum were studied for antimicrobial and cytotoxic activities. Argyranthemum adauctum, A. foeniculaceum and A. frutescens showed antimicrobial activity against Gram-positive and Gram-negative and cytotoxic activity against HeLa and Hep-2 cell lines. Two new acetylenic compounds, frutescinol isovalerate and 3'-demethyl frutescinol isovalerate, were isolated from A. frutescens and their structures elucidated by spectroscopic studies. PMID- 9214779 TI - Inversion of the current-distance relationship by transient depolarization. AB - The objective of this research was to develop a technique to excite selectively nerve fibers distant from an electrode without exciting nerve fibers close to the electrode. The shape of the stimulus current waveform was designed based on the nonlinear conductance properties of neuronal sodium channel. Models of mammalian peripheral myelinated axons and experimental measurements on cat sciatic nerve were used to determine the effects of subthreshold polarization on neural excitability and recruitment. Subthreshold membrane depolarization generated a transient decrease in neural excitability and thus an increase in the threshold for stimulation by a subsequent stimulus pulse. The decrease in excitability increased as the duration and amplitude of the subthreshold depolarization were increased, and the increase in threshold was greater for fibers close to the electrode. When a depolarizing stimulus pulse was applied immediately after the subthreshold depolarization, nerve fibers far from the electrode could be stimulated without stimulating fibers close to the electrode. Subthreshold depolarizing prepulses inverted the current-distance relationship and allowed selective stimulation of nerve fibers far from the electrode. PMID- 9214778 TI - Two quinoline alkaloids from the Caribbean cyanobacterium Lyngbya majuscula. AB - Fractionation of the lipid extract of the marine cyanobacterium Lyngbya majuscula collected from Curacao afforded two quinoline alkaloids in low yield. Their structures were determined as 4,8-dimethyl-6-O-(2'-4'-di-O-methyl-beta-D xylopyranosyl)-hydroxyquinoli ne and 4,8-dimethyl-6-hydroxyquinoline on the basis of spectroscopic analysis, mainly 2D NMR spectroscopy. PMID- 9214780 TI - The transient far-field response of a discontinuous one-dimensional cardiac fiber to subthreshold stimuli. AB - It is hypothesized that the discontinuous structure of cardiac tissue is at least partly responsible for the ability of extracellular field stimuli to affect those regions of cardiac tissue very far from the stimulating electrodes. Yet, the details of this "far-field" interaction are still poorly understood. This paper derives analytical closed-form equations that describe the far-field spatial and temporal behaviors of the axial currents and transmembrane voltages along a discontinuous cardiac fiber in response to any applied subthreshold stimulus. Moreover, these derivations incorporate the influences of both junctional resistance and postulated junctional capacitance on such responses. Thus, as compared with previously-derived techniques, these equations extend and simplify the generation and analysis of such far-field responses. Frequency analysis of this system demonstrates that the fiber generally behaves as a low-pass filter, with the location of its corner frequency highly dependent on the magnitudes of both junctional resistance and especially junctional capacitance. Increasing either junctional resistance or capacitance significantly and monotonically decreases the value of the corner frequency--equivalently manifest as a large increase in the duration of the transient response to a step input stimulus. Such changes to these initial excitation dynamics might prove relevant during the far field stimulation of cardiac tissue, such as during defibrillation-type shocks. PMID- 9214781 TI - A new method for regularization parameter determination in the inverse problem of electrocardiography. AB - Computing the potentials on the heart's epicardial surface from the body surface potentials constitutes one form of the inverse problem of electrocardiography. An often-used approach to overcoming the ill-posed nature of the inverse problem and stabilizing the solution is via zero-order Tikhonov regularization, where the squared norms of both the surface potential residual and the solution are minimized, with a relative weight determined by a so-called regularization parameter. This paper looks at the composite residual and smoothing operator (CRESO) and L-curve methods currently used to determine a suitable value for this regularization parameter, t, and proposes a third method that works just as well and is much simpler to compute. This new zero-crossing method selects t such that the squared norm of the surface potential residual is equal to t times the squared norm of the solution. Its performance was compared with those of the other two methods, using three stimulation protocols of increasing complexity. The first of these protocols involved a concentric spheres model for the heart and torso and three current dipoles placed inside the inner sphere as the source distribution. The second replaced the spheres with realistic epicardial and torso geometries, while keeping the three-dipole source configuration. The final simulation kept the realistic epicardial and torso geometries, but used epicardial potential distributions corresponding to both normal and ectopic activation of the heart as the source model. Inverse solutions were computed in the presence of both geometry noise, involving assumed erroneous shifts in the heart position, and of Gaussian measurement noise added to the torso surface potentials. It was verified that in an idealistic situation, in which correlated geometry noise dominated the uncorrelated Gaussian measurement noise, only the CRESO approach arrived at a value for t. Both L-curve and zero-crossing approaches did not work. Once measurement noise dominated geometry noise, all three approaches resulted in comparable t values. It was also shown, however, that often under low measurement noise conditions none of the three resulted in an optimum solution. PMID- 9214782 TI - Magnetoencephalography with diversely oriented and multicomponent sensors. AB - To locate endocranial current sources, a magnetoencephalography (MEG) system usually measures the magnetic field at many points around the skull with an array of radial sensors. Despite the success of using radial components of the field, we show that using nonradial components may potentially also be beneficial. We demonstrate some benefits of using diversely oriented and multicomponent sensors to measure the nonradial components. A framework is provided for analyzing the accuracy of a system that estimates the location and direction of a current dipole inside a spherical skull. The framework is then used to determine the effect on accuracy of varying the orientations of sensors in an array and, as a consequence, it is found that the radial orientations commonly used in practice are suboptimal for locating dipoles near the array's center. A diversely oriented array that improves performance is presented. We show how a single multicomponent sensor can locate a dipole, and derive a simple algorithm for locating a dipole near the sensor. PMID- 9214783 TI - Fetal ECG extraction from single-channel maternal ECG using singular value decomposition. AB - The extraction of fetal electrocardiogram (ECG) from the composite maternal ECG signal obtained from the abdominal lead is discussed. The proposed method employs singular value decomposition (SVD) and analysis based on the singular value ratio (SVR) spectrum. The maternal ECG (M-ECG) and the fetal ECG (F-ECG) components are identified in terms of the SV-decomposed modes of the appropriately configured data matrices, and elimination of the M-ECG and determination of F-ECG are achieved through selective separation of the SV-decomposed components. The unique feature of the method is that only one composite maternal ECG signal is required to determine the F-ECG component. The method is numerically robust and computationally efficient. PMID- 9214784 TI - Estimating alertness from the EEG power spectrum. AB - In tasks requiring sustained attention, human alertness varies on a minute time scale. This can have serious consequences in occupations ranging from air traffic control to monitoring of nuclear power plants. Changes in the electroencephalographic (EEG) power spectrum accompany these fluctuations in the level of alertness, as assessed by measuring simultaneous changes in EEG and performance on an auditory monitoring task. By combining power spectrum estimation, principal component analysis and artificial neural networks, we show that continuous, accurate, noninvasive, and near real-time estimation of an operator's global level of alertness is feasible using EEG measures recorded from as few as two central scalp sites. This demonstration could lead to a practical system for noninvasive monitoring of the cognitive state of human operators in attention-critical settings. PMID- 9214785 TI - The inaccuracy of Kubicek's one-cylinder model in thoracic impedance cardiography. AB - The validity of a one- and a two-cylinder model, underlying thoracic impedance cardiography (TIC), was investigated by studying the length dependence of the impedance parameters Z0, (dZ/dt)min, and stroke volume (SV). It can be shown that, within a one-cylinder model, all parameters are directly proportional to the length, whereas, if the volume conduction of the thorax and the neck are modeled separately, Z0 and (dZ/dt)min are expected to be linear dependent and SV will be nonlinear upon the length. The expectations were compared to results from in vivo measurements. Two electrode arrays were studied, in which the caudal recording electrode position was varied; SV was calculated using Kubicek's equation. Except for small distances, the results showed a nearly linear relation between the parameters and the length. Regression analysis of the linear part revealed statistically significant intercepts (p < 0.05). Neither the intercept nor the nonlinear part can be explained by a one-cylinder model, whereas a model consisting of two cylinders serially connected describes the experimental results accurately. Thus SV estimation based on a one-cylinder model is biased due to the invalid one-cylinder model. Corrections for the Kubicek-equation need to be developed in future research using this two-cylinder model. PMID- 9214786 TI - Development of a microcontroller-based automatic control system for the electrohydraulic total artificial heart. AB - An automatic physiological control system for the actively filled, alternately pumped ventricles of the volumetrically coupled, electrohydraulic total artificial heart (EHTAH) was developed for long-term use. The automatic control system must ensure that the device: 1) maintains a physiological response of cardiac output, 2) compensates for an nonphysiological condition, and 3) is stable, reliable, and operates at a high power efficiency. The developed automatic control system met these requirements both in vitro, in week-long continuous mock circulation tests, and in vivo, in acute open-chested animals (calves). Satisfactory results were also obtained in a series of chronic animal experiments, including 21 days of continuous operation of the fully automatic control mode, and 138 days of operation in a manual mode, in a 159-day calf implant. PMID- 9214787 TI - Analog transmission line model for simulation of systemic circulation. AB - A simple four-tube arteries-microvessels-veins system which simulates a more realistic loading for human circulation was built using transmission line network. Hemodynamic data from literature are used in the fluid-circuit analogy, and the flow leakage and viscoelastic properties of the blood vessels have been considered. The effect of veins on the input impedance spectrum was found to be negligibly small above 0.5 Hz. The predicted input impedance spectra agree reasonably well with the published measurements both in shape and magnitude. Parametric analysis shows that the changes of vascular properties in the lower body affect the first minimum, and the changes in the upper body influence the second minimum. The blood flow in and out of kidney and liver dominates the aortic impedance from 0 to 5 Hz. Decreasing capacitance (i.e., increasing arterial stiffness due to aging), reducing the lumen area, or decreasing the length of blood vessels result in an increase in the impedance modulus, and the first minimum shift to a higher frequency which agree well with experiments. In the current model, the pressure, flow waveform, and local impedance can be predicted at any location along the circulatory tree. The characteristic of arterial pulse propagation resembles published measurements. PMID- 9214788 TI - A sampling theorem for EEG electrode configuration. AB - An analytical tool to help in selecting the number of electrodes required for recording electroencephalogram (EEG) signals is presented. The main assumption made is that the scalp can be modeled as a hemispherical surface. The number of sensors required to sample a surface is derived by using a mean square error (MSE) measure to approximate the continuous potential functions on the hemispherical surface. An algorithm for selecting the number of electrodes for arbitrary head geometries is also proposed. A sampling theorem is then derived with conditions on the sampling points for electrode placement. PMID- 9214789 TI - Maximum entropy method for magnetoencephalography. AB - Simulations show that the maximum entropy method is a promising technique for image reconstruction in magnetoencephalography. An algorithm based on the work of Skilling and Bryan [1] and an appropriately modified expression for the "entropy" is shown to provide high-quality reconstructions of both isolated and dense distributions of neural current "dipoles", neglecting return currents. In particular the results are substantially superior to those obtained with the well known minimum norm procedure. PMID- 9214790 TI - EEG data compression techniques. AB - In this paper, electroencephalograph (EEG) and Holter EEG data compression techniques which allow perfect reconstruction of the recorded waveform from the compressed one are presented and discussed. Data compression permits one to achieve significant reduction in the space required to store signals and in transmission time. The Huffman coding technique in conjunction with derivative computation reaches high compression ratios (on average 49% on Holter and 58% on EEG signals) with low computational complexity. By exploiting this result a simple and fast encoder/decoder scheme capable of real-time performance on a PC was implemented. This simple technique is compared with other predictive transformations, vector quantization, discrete cosine transform (DCT), and repetition count compression methods. Finally, it is shown that the adoption of a collapsed Huffman tree for the encoding/decoding operations allows one to choose the maximum codeword length without significantly affecting the compression ratio. Therefore, low cost commercial microcontrollers and storage devices can be effectively used to store long Holter EEG's in a compressed format. PMID- 9214792 TI - Recognition of temporally changing action potentials in multiunit neural recordings. AB - We present a method to iteratively train an artificial neural network (ANN) or other supervised pattern classifier in order to adaptively recognize and track temporally changing patterns. This method uses recently acquired data and the existing classifier to create new training sets, from which a new classifier is then trained. The procedure is repeated periodically using the most recently trained classifier. This scheme was evaluated by applying it to simulated situations that arise in chronic recordings of multiunit neural activity from peripheral nerves. The method was able to track the changes in these simulated chronic recordings and to provide better unit recognition rates than an unsupervised clustering method suited to this problem. PMID- 9214791 TI - A patient-specific algorithm for the detection of seizure onset in long-term EEG monitoring: possible use as a warning device. AB - During long-term electroencephalogram (EEG) monitoring of epileptic patients, a seizure warning system would allow patients and observers to take appropriate precautions. It would also allow observers to interact with patients early during the seizure, thus revealing clinically useful information. We designed patient specific classifiers to detect seizure onsets. After a seizure and some nonseizure data are recorded in a patient, they are used to train a classifier. In subsequent monitoring sessions, EEG patterns have to pass this classifier to determine if a seizure onset occurs. If it does, an alarm is triggered. Extreme care has been taken to ensure a low false-alarm rate, since a high false-alarm rate would render the system ineffective. Features were extracted from the time and frequency domains and a modified nearest-neighbor (NN) classifier was used. The system reached an onset detection rate of 100% with an average delay of 9.35 a after onset. The average false-alarm rate was only 0.02/h. The method was evaluated in 12 patients with a total of 47 seizures. Results indicate that the system is effective and reasonably reliable. Computation load has been kept to a minimum so that real-time processing is possible. PMID- 9214793 TI - Late potential recognition by artificial neural networks. AB - Ventricular late potentials (LP's) are high-frequency low-amplitude signals obtained from signal-averaged electrocardiograms (ECG's) [SAECG's]. LP's are useful in identifying patients prone to ventricular tachycardia (VT), spontaneous or inducible during electrophysiology testing. A combination of self-organizing and supervised artificial neural network (ANN) models was developed to identify patients with a positive electrophysiology (PEP) test for inducible ventricular tachycardia from patients with a negative electrophysiology (NEP) test using LP's. We have added morphology information of vector magnitude waveform to original set of three time-domain features of LP's, which are total QRS duration (TQRSD), high-frequency low-amplitude signal duration (HFLAD), and root-mean square voltage (RMSV). Pattern recognition results from an ANN model with this combination feature set are superior to the results from Bayesian classification model based on conventional three time-domain features of SAECG. In order to increase the robustness of the recognition, a filtered QRS offset point is randomly shifted +/- 8 ms to form a fuzzy training set, which was to simulate the possible error in detecting QRS offset point of filtered SAECG. We also found that nonlinear transformation through the hidden layer of developed ANN model could increase Euclidean distance between PEP and NEP patterns. PMID- 9214795 TI - Identifying and tracking a guide wire in the coronary arteries during angioplasty from X-ray images. AB - During angioplasty, a guide wire (GW) is routinely placed in the coronary artery. Balloon inflation during angioplasty causes transient occlusion of the coronary artery and regional dysfunction. Thus, it is of major importance to monitor myocardial function, which may be impaired during this period. Since the GW moves with the coronary arteries, information regarding myocardial function can potentially be extracted from the GW motion. An algorithm is suggested which is a step toward such monitoring. The algorithm presented is a semiautomatic procedure for identifying and tracking the GW using specific characteristics of the GW. This algorithm is based on working in limited active windows. A preprocessing stage which enhances the GW by the use of a modified Laplacian filter or a modified Marr-Hildreth filter is introduced. The second stage of the algorithm is the tracking of the GW, which is based on fitting a second-degree polynomial to the GW using the Hough transform in each window. To further improve the results further modifications of the basic algorithms that were taken. A single set of parameters, which enabled good tracking for a large number of images taken during angioplasty, was fitted to the final algorithm. PMID- 9214794 TI - Cylindrical ultrasonic transducers for cardiac catheter ablation. AB - This study was designed to evaluate the feasibility of using cylindrical ultrasound transducers mounted on a catheter for the ablation of cardiac tissues. In addition, the effects of ultrasound frequency and power was evaluated both using computer simulations and in vitro experiments. Frequencies of 4.5, 6, and 10 MHz were selected based on the simulation studies and manufacturing feasibility. These transducers were mounted on the tip of 7-French catheters and applied in vitro to fresh ventricular canine endocardium, submerged in flowing degassed saline at 37 degree C. When the power was regulated to maintain transducer interface temperature at 90-100 degree C, the 10-, 6-, and 4.5-MHz transducers generated a lesion depth of 5.9 +/- 0.2 mm, 4.6 +/- 1.0 mm, and 5.3 +/- 0.9 mm, respectively. The 10-MHz transducer was chosen for the in vivo tests since the maximum lesion depth was achieved with the lowest power. Two dogs were anesthetized and sonications were performed in both the left and right ventricles. The 10-MHz cylindrical transducers caused an average lesion depth of 6.4 +/- 2.5 mm. In conclusion, the results show that cylindrical ultrasound transducers can be used for cardiac tissue ablation and that they may be able to produce deeper tissue necrosis than other methods currently in use. PMID- 9214796 TI - Role of intrinsic muscle properties in producing smooth movements. AB - Human upper limb movement trajectories have been shown to be quite smooth, in that time derivatives of end point position (r), including d3r/dt3 (i.e., jerk), appear to be minimized during rapid voluntary reaching tasks. Studies have suggested that these movements are implemented by an optimal neural controller which seeks to minimize a cost function, such as average jerk cost, over the course of these motions. While this hypothetical control strategy is widely supported, there are substantial difficulties associated with implementing such a controller, including ambiguities inherent in transformations from Cartesian to joint coordinates, and the lack of appropriate transducers to provide information about higher derivatives of limb motion to the nervous system. Given these limitations, we evaluate the possibility that smoothing of movement might be induced primarily by the intrinsic mechanical properties of muscle by recording the trajectories of inertially loaded muscle with the excitatory input held constant. These trajectories are compared with those predicted by a minimum-jerk optimization model, and by a Hill-based muscle model. Our results indicate that trajectories produced by inertially loaded muscle alone are smooth (in the minimum-jerk sense), and that muscle properties may suffice to account for much of the observed smoothing of voluntary motion, obviating the need for an optimizing neural strategy. PMID- 9214797 TI - Theoretical study of magnetic field of current monopoles in special volume conductor using symmetry analysis. AB - We derive a formula for the magnetic field outside volume conductors having axial symmetry with radial and axial symmetrically distributed source currents. The magnetic field is shown to have components only along the cylindrical polar angle direction and its magnitude to depend only on the topological structure of the volume conductor and the location of the source current. With this formula, the magnetic field generated by the volume current of a current monopole within and on the symmetrical axis of several volume conductors (such as semi-infinite volume, infinite slab, sphere, infinite cylinder, semi-infinite cylinder, finite cylinder, prolate spheroid, and oblate spheroid) is shown to be equivalent to the magnetic field generated by a line current calculated using the Biot-Savart's law. In the first three volume conductors, the monopole solution of the magnetic field allows the calculation of magnetic fields generated by arbitrarily distributed (and balanced for finite volume conductors) current monopoles. PMID- 9214798 TI - Regional regularization of the electrocardiographic inverse problem: a model study using spherical geometry. AB - This study examines the use of a new regularization scheme, called regional regularization, for solving the electrocardiographic inverse problem. Previous work has shown that different time frames in the cardiac cycle require varying degrees of regularization. This reflects differences in potential magnitudes, gradients, signal-to-noise ratio (SNR), and locations of electrical activity. One might expect, therefore, that a single regularization parameter and a uniform level of regularization may also be insufficient for a single potential map of a single time frame because in one map there are regions of high and low potentials and potential gradients. Regional regularization is a class of methods that subdivides a given potential map into functional "regions" based on the spatial characteristics of the potential ("spatial frequencies"). These individual regions are regularized separately and recombined into a complete map. This paper examines the hypothesis that such regionally regularized maps are more accurate than if all regions were taken together and solved with an averaged level of regularization. In a homogeneous concentric spheres model, Legendre polynomials are used to decompose a torso potential map into a set of submaps, each with a different degree of spatial variation. The original torso map is contaminated with data noise, or geometrical error or both, and regional regularization improves the epicardial potential reconstruction by up to 25% [relative error (RE)]. Regional regularization also improves the reconstructed location of peaks. A practical goal is to extend the application of this method to the realistic torso geometry, but because Legendre decomposition is limited to geometries with spherical symmetry, other methods of map decomposition must be found. Singular value decomposition (SVD) is used to decompose the maps into component parts. Its individual submaps also have different levels of spatial variation; moreover, it is generalizable to any vector, does not require spherical symmetry, and is extremely efficient numerically. Using SVD decomposition for regional regularization, significant improvement was achieved in the map quality in the presence of data noise. PMID- 9214799 TI - Three-dimensional finite-difference bidomain modeling of homogeneous cardiac tissue on a data-parallel computer. AB - In this paper, a data-parallel computer is used to provide the memory and reduction in computer time for solving large finite-difference bidomain problems. The finite-difference grid is mapped effectively to the processors of the parallel computer, simply by mapping one node to one (virtual) processor. Implemented on the connection machines (CM's) CM-200 and CM-5, the data-parallel finite-difference algorithm has allowed the solution of finite-difference bidomain problems with over 2 million nodes. Details on the algorithm are presented together with computational performance results. PMID- 9214800 TI - A silicon bidirectional flow sensor for measuring respiratory flow. AB - We describe a solid-state, silicon integrated, bidirectional flow sensor for respiratory applications. The sensor is a thermal vector sensor. The electronic circuitry for obtaining bidirectional sensitivity is presented together with actual application to a healthy volunteer put on mechanical ventilation. The sensor's input flow range is from -60 to +60 L/min, and its rise-time is < or = 40 ms and fall-time is < or = 60 ms. The effect of changes in gas composition as used in mechanically ventilated patients on the sensor output signal are estimated to be less than 2%. The temperature sensitivity is about -1.5% per degree Celsius. PMID- 9214801 TI - The inverse problem in electroretinography: a study based on skin potentials and a realistic geometry model. AB - The problem of obtaining the retinal source distribution that generates the electroretinogram (ERG) from measured skin potentials is addressed. A realistic three-dimensional (3-D) volume conductor model of the head is constructed from magnetic resonance image (MRI) data sets. The skin potential distribution generated in this model by a dipole layer source at the retina is computed by using the boundary element method (BEM). The influence of the various compartments of the complete model on the results was investigated, and a simplified model was defined. An inverse procedure for estimating the source distribution at the retina from ERG's obtained from skin electrodes was developed. The procedure was tested on simulated potentials. A fair correspondence between the original and estimated source distribution was found. Furthermore, the ERG's measured at seven skin electrodes were used to estimate the source distribution at the retina. The ERG potential waveform at an additional skin electrode was computed from this source distribution and compared to the measured potential at this electrode. Again a fair correspondence was obtained. It is concluded that the methods may become a useful tool for clinical applications, i.e., for the assessment of localized defects in retinal function. PMID- 9214802 TI - Microwave tomography: two-dimensional system for biological imaging. AB - Microwave tomographic imaging is one of the new technologies which has the potential for important applications in medicine. Microwave tomographically reconstructed images may potentially provide information about the physiological state of tissue as well as the anatomical structure of an organ. A two dimensional (2-D) prototype of a quasi real-time microwave tomographic system was constructed. It was utilized to reconstruct images of physiologically active biological tissues such as an explanted canine perfused heart. The tomographic system consisted of 64 special antennae, divided into 32 emitters and 32 receivers which were electronically scanned. The cylindrical microwave chamber had an internal diameter of 360 mm and was filled with various solutions, including deionized water. The system operated on a frequency of 2.45 GHz. The polarization of the incident electromagnetic field was linear in the vertical direction. Total acquisition time was less than 500 ms. Both accurate and approximation methods of image reconstruction were used. Images of 2-D phantoms, canine hearts, and beating canine hearts have been achieved. In the worst-case situation when the 2-D diffraction model was used for an attempt to "slice" three dimensional (3-D) object reconstruction, we still achieved spatial resolution of 1 to 2 cm and contrast resolution of 5%. PMID- 9214803 TI - Microwave imaging for tissue assessment: initial evaluation in multitarget tissue equivalent phantoms. AB - A prototype microwave imaging system is evaluated for its ability to recover two dimensional (2-D) electrical property distributions under transverse magnetic (TM) illumination using multitarget tissue equivalent phantoms. Experiments conducted in a surrounding lossy saline tank demonstrate that simultaneous recovery of both the real and imaginary components of the electrical property distribution is possible using absolute imaging procedures over a frequency range of 300-700 MHz. Further, image reconstructions of embedded tissue-equivalent targets are found to be quantitative not only with respect to geometrical factors such as object size and location but also electrical composition. Quantitative assessments based on full-width half-height criteria reveal that errors in diameter estimates of reconstructed targets are less than 10 mm in all cases, whereas, positioning errors are less than 1 mm in single object experiments but degrade to 4-10 mm when multiple targets are present. Recovery of actual electrical properties is found to be frequency dependent for the real and imaginary components with background values being typically within 10-20% of their correct size and embedded object having similar accuracies as a percentage of the electrical contrast, although errors as high as 50% can occur. The quantitative evaluation of imaging performance has revealed potential advantages in a two-tiered receiver antenna configuration whose measured field values are more sensitive to target region changes than the typical tomographic type of approach which uses reception sites around the full target region perimeter. This measurement strategy has important implications for both the image reconstruction algorithm where there is a premium on minimizing problem size without sacrificing image quality and the hardware system design which seeks to economize on the amount of measured data required for quantitative image reconstruction while maximizing its sensitivity to target perturbations. PMID- 9214804 TI - Unwrapping Cochlear implants by spiral CT. AB - Multielectrode, intracochlear implants were designed for individuals with profound sensorineural hearing loss who derive little or no benefit form acoustic hearing aids. Determination of each electrode's position in a patient's inner ear may improve speech processor programming to maximize speech recognition. In this paper, an approach is described to use as input a volumetric spiral computed tomography (CT) image of the Nucleus electrode array (Cochlear Pty. Ltd, Lane Cove, NSW, Australia) to unwrap it, and to measure its implanted length given starting and end points. Representative curvilinear structures were digitally synthesized in image volumes of isotropic 0.1-mm voxels. The electrode array was spirally CT-scanned in vitro and in vivo, and reconstructed on an isotropic grid in 0.1-mm steps. Two algorithms were constructed to track and measure these curvilinear structures. The first algorithm is Karhunen-Loeve (K-L)-transform based, in which the K-L transform is locally applied at a current main axis position to determine the eigenvectors of the main axis voxels, the next main axis position is estimated from the current position along the principal eigendirection, adjusted to the mass center of the orthogonal cross section passing through the estimated position, and then scaled to have a prespecified step. The second algorithm is similar to the first one but avoids use of the K-L transform. In the second algorithm, the next position is directly estimated along the local direction and then processed with the same correction and scaling operations. With user-specified starting and end points as well as a local direction at the starting point, a curvilinear structure can be automatically tracked using either of the algorithms. The first algorithm is more robust, while the second one is more efficient. In the numerical and in vitro studies, the lengths of the curvilinear structures were accurately measured. Given local directions determined in the tracking process, an electrode array image can be unwrapped into a linear array with the central electrode axis as the abscissa. The unwrapping approach allows longitudinally and cross-sectionally accurate measurement and better visualization of cochlear implant images. With preimplantation knowledge of length, width, and center electrode distance, the position of individual electrodes can be estimated after unwrapping. PMID- 9214805 TI - A model of ultrasound backscatter for the assessment of myocardial tissue structure and architecture. AB - A statistical parametric model of returning echoes from myocardium is theorized in order to investigate the relationship between normal myocardium structure and spectral signatures with the use of ultrasonic tissue characterization. It is hypothesized, that in a clinical setting the normal myofiber architecture in the left ventricular wall is structured as a matrix of cylinder scatterers whose orientation and spatial distribution vary according to two different statistical distribution laws: 1) a Gaussian law to approximate parametric angular myofiber variability at each site within the myocardial wall; 2) a gamma distribution law to describe parametric regularity in scatterer interdistance. In the model, the effect of the angle of insonification with respect to the alignment of myofibers on ultrasound backscatter was considered. The slope of the power spectral density (PSD) evaluated within the echocardiographic transducer bandwidth has been used as a ultrasonic tissue characterization parameter. The model has been tested by computer simulation and in vitro measurements on myocardial pig tissue specimens. The concordance between experimental and simulated results confirms that the model accounts for the process underlying the echo formation from normal myocardium. Moreover, it provides a simple method of simulation which can be easily implemented and used for the assessment of pathologic alterations. PMID- 9214806 TI - An ultrasound indentation system for biomechanical properties assessment of soft tissues in-vivo. AB - An ultrasound indentation system for biomechanical assessment of soft tissues in vivo was developed. The pen-size, hand-held probe was composed of an ultrasound transducer and a load cell. The ultrasound transducer was at the tip of the probe serving also as the indentor. The thickness and deformation of the soft tissue layer were determined from the ultrasound echo. A compressive load cell was connected in series with the ultrasound transducer to record the force response. A validation experiment was performed on porcine tissues. Force and deformation acquired with the present system was in good comparison with those obtained from a Housfield material testing machine. Material constants were obtained via a curve-fitting procedure by predicting the force transient response from the deformation-time data using a quasilinear viscoelastic model. In addition, deformation in the fat and in the muscle could be differentiated. The potential applications of this type of indentation probes are many. The specific application of this current development is for stump tissue assessment in the design of prosthetics. PMID- 9214807 TI - Improved estimation of low velocities in color Doppler imaging by adapting the mean frequency estimator to the clutter rejection filter. AB - An adaptive mean frequency estimator is proposed for color flow imaging. It is based on a series expansion of the first derivative of the autocorrelation function of the Doppler signal at origin. Its bias can be reduced by shifting the integration bounds in the series expansion and its variance adjusted by adapting the coefficients of the serial development. This estimator can be fitted to the specific characteristics of the clutter rejection filter using the signal-to noise ratio (SNR) of the Doppler signal as an adaptive parameter. Its performance is compared to that of the usual correlation angle estimator, and its thresholded version, as well as that of the general mean frequency estimator, using a model of Doppler signal. The detection of low frequencies was significantly improved. The mean square error (MSE) was reduced an average 15 fold over a 25-dB range on the SNR, compared to the correlation angle estimator (CAE) or the general mean frequency estimator. A two-fold reduction in the MSE was obtained compared to the thresholded correlation angle estimator. PMID- 9214808 TI - Segmentation of digital signals based on estimated compression ratio. AB - We have studied the problem of approximating a digital signal with a suitable continuous broken line. We use the approximative broken line for further analysis of the signal as detection of peaks, waves, and other structural features. We can also save considerable amount of storage space with an approximation that does not lose too much significant information about the original signal. Our work is based on examining different distance metrics and different segmentation methods with respect to the remaining residual error in the resulting approximation. The aim of the work has been to develop a method that can perform segmentation with an acceptable amount of residual error without a need to define a large set of parameters that control the segmentation process. Our contribution is to examine the effect of the estimated compression ratio of the resulting approximation and finding an estimate of this compression ratio. We first define a target in the form of a compression ratio of the resulting approximation and then by applying our method, try to find a suitable threshold parameter to achieve this target. We have tested our method with electrocardiogram (ECG) signals and the compression ratio of the approximation has been found to be a suitable target to control the segmentation process. PMID- 9214809 TI - Potential and current density distributions of cranial electrotherapy stimulation (CES) in a four-concentric-spheres model. AB - Cranial electrotherapy stimulation (CES) has been successfully used for treatment of many psychiatric diseases. Its noninvasive nature is its major advantage over other forms of treatments such as drugs. It is postulated that the low electric current of CES causes the release of neurotransmitters. However, the current pathways have not been extensively investigated. In the following paper, analytical and numerical methods are used to determine the distribution of potential and current density in a four zone concentric spheres model of the human head when excited by two electrodes diametrically opposite to each other. Because of the azimuthal symmetry, which is assumed in this study, a two dimensional (2-D) finite difference approximation is derived in the spherical grid. The current density distribution is projected around the center of the model, where the thalamus is modeled as a concentric sphere. All dimensions and electrical properties of the model are adapted from clinical data. Results of this simulation indicate that, in contrast to previous beliefs, a small fraction of the CES current does reaches the thalamic area and may facilitate the release of neurotransmitters. PMID- 9214810 TI - Micrometer resolution silane-based patterning of hippocampal neurons: critical variables in photoresist and laser ablation processes for substrate fabrication. AB - Toward the goal of creating patterns of primary hippocampal neurons in low density culture, we investigated techniques to fabricate microminiature grids of organofunctional silanes on glassy surfaces. A new photoresist (PR) process, Selective Silane Removal (SSR), was developed and compared to two previously developed techniques which use PR and laser patterning. The grid patterns consisted of 27 combinations of path width, length, and intersection (node diameter). The background consisted of squares bounded by the paths. The best neuron patterning was observed on substrates produced by the SSR process where cytophilic aminosilane is uniformly deposited and selectively removed from the background. Controlling water during aminosilane deposition was critical to good neuronal growth and patterning. Oxygen plasma etching of background regions prior to cytophobic phenylsilane binding significantly reduced off-pattern cell growth. Up to 90% of somata grown on these substrates complied to the pattern, and an average of 77% of background regions were free of neurites or cells connected to the pattern. The highest laser energy density, 120 mJ/cm2, produced the best compliance on lased substrates, with an average of 35% of background regions free of connected cells and neurites, but considerable variation across the surface. On substrates with excellent patterning, compliance to nodes was found to be dependent on pattern dimensions, with 20-micron node diameters and 80-micron internodal path lengths increasing compliance. PMID- 9214811 TI - Measures of postural steadiness: differences between healthy young and elderly adults. AB - Measures of postural steadiness are used to characterize the dynamics of the postural control system associated with maintaining balance during quiet standing. The objective of this study was to evaluate the relative sensitivity of center-of-pressure (COP)-based measures to changes in postural steadiness related to age. A variety of time and frequency domain measures of postural steadiness were compared between a group of twenty healthy young adults (21-35 years) and a group of twenty healthy elderly adults (66-70 years) under both eyes-open and eyes-closed conditions. The measures that identified differences between the eyes open and eyes-closed conditions in the young adult group were different than those that identified differences between the eye conditions in the elderly adult group. Mean velocity of the COP was the only measure that identified age-related changes in both eye conditions, and differences between eye conditions in both groups. The results of this study will be useful to researchers and clinicians using COP-based measures to evaluate postural steadiness. PMID- 9214812 TI - Reproducibility of MEG auditory evoked field source localizations in normal human subjects using a seven-channel gradiometer. AB - Magnetoencephalographic (MEG) auditory evoked fields (EF) were recorded from 12 normal adult subjects over both hemispheres on two separate occasions at least one week apart using a seven-channel second-order gradiometer. Stimuli were computer-generated at 25-msec duration, 1 kHz tone pips. Responses to 100 stimuli were averaged, and source estimates with confidence intervals were computed, for the 100-msec latency auditory EF component, termed M100. Root-mean-squared (rms) differences in x, y, and z locations were approximately 0.7 cm on the two occasions; strength and orientation differences were 18 nA-m and 11 degrees, respectively. This spatial accuracy using a seven-channel instrument, compares favorably with other currently available technologies for localization of brain function. PMID- 9214813 TI - Potential distribution for a spheroidal cell having a conductive membrane in an electric field. AB - When a cell is situated in a uniform electric field, the field is modified due to the relatively low conductance of the cell membrane compared to that of the surrounding fluids. In certain cases, such as in the estimation of internal and external electrokinetic forces, one requires a means of estimating the magnitude of the electric field inside and outside the cell. Most treatments consider the case when the membrane has zero conductivity, or the case of only a spherical cell. We solve Laplace's equation for the electric potential distribution inside and outside a cell having a prolate spheroidal shape and having a membrane with a finite, nonzero conductivity. PMID- 9214814 TI - Estimation of single event-related potentials utilizing the Prony method. AB - This paper deals with estimation of the waveform of a single event-related potential, sERP. An additive noise model is used for the measured signal and the SNR of the disturbed sERP is approximately 0 dB. The sERP is described by a series expansion where the basis functions are damped sinusoids. The fundamental basis function is estimated by the least squares Prony method, derived for colored noise. The performance of the Prony method for different forms of the power density spectrum of the noise is investigated. A white noise approximation can be used at a low signal-to-noise (SNR). The basis functions change slowly but the waveform of the sERP may vary from one stimulus to another, thus we average a small number of correlation functions in order to increase the SNR. The method is evaluated by using measurements from four subjects and the results confirm the variability of the sERP. PMID- 9214815 TI - Modeling of cardiovascular variability using a differential delay equation. AB - The influence of time delay in the baroreflex control of the heart activity is analyzed by using a simple mathematical model of the short-term pressure regulation. The mean arterial pressure in a Windkessel model is controlled by a nonlinear feedback driving a nonpulsatile model of the cardiac pump in accordance with the steady-state characteristics of the arterial baroreceptor reflex. A pure time delay is placed in the feedback branch to simulate the latent period of the baroreceptor regulation. Because of system nonlinearity model dynamics is found to be highly sensitive to time delay and changes of this parameter within a physiological range cause the model to exhibit different patterns of behavior. For low values of time delay (shorter than 0.5 s) the model remains in a steady state. When time delay is longer than 0.5 s, a Hopf bifurcation is crossed and spontaneous oscillations occur with frequencies in the high-frequency (HF) band. Further increases of time delay above 1.2s cause the oscillations to become more complex, and following the typical Feigenbaum cascade, the system becomes chaotic. In this condition heart rate, and flow show evident variability. The heart rate power spectrum exhibits a peak whose frequency moves from the HF to LF band depending on whether simulated time delay is as short as the vagal-mediated control or long as the sympathetic one. PMID- 9214816 TI - Extraction of spatio-temporal signatures from depth EEG seizure signals based on objective matching in warped vectorial observations. AB - In the field of epilepsy, the analysis of stereoelectroencephalographic (SEEG) signals recorded with depth electrodes provides major information on interactions between brain structures during seizures. A comprehensive methodology of comparing SEEG seizure recordings is presented. It proceeds in three steps: 1) segmentation of SEEG signals; 2) characterization and labeling of segments; and 3) comparison of observations coded as sequences of symbol vectors. The third step reports a vectorial extension of the Wagner and Fischer's algorithm to first, quantify similarities between observations and second, extract invariant sequences of events, referred to as spatiotemporal signatures. The study shows that two observations of nonequal duration can be matched by deforming the first one to optimally fit the second, under cost constraints. Results show that the methodology allows to exhibit signatures occurring during epileptic seizures and to point out different types of seizure patterns. The study brings objective results on reproducible interactions between brain structures during ictal periods and may help in the understanding of epileptogenic networks. PMID- 9214817 TI - A technique for removal of the visuoperceptual component from tracking performance and its application to Parkinson's disease. AB - Although it is well established that subjects with Parkinson's disease perform poorly on complex sensory-motor tasks, the extent to which this is due to visuoperceptual deficits is unclear. We measured the performance of 16 patients with Parkinson's disease, both on and off drugs, and 16 age and sex matched control subjects on preview and nonpreview tracking tasks and a nonmotor test of dynamic visuoperception. Order effects were controlled for by a randomized cross over design. Performance on the perceptual task was measured in terms of perceptual resolution and was found impaired in the Parkinsonian group. The contribution of visuoperceptual function to tracking performance was removed using the concept of a visuoperceptual buffer-zone. The mean tracking error remained impaired on all tracking tasks and demonstrated that limitations is visuoperceptual function play only a minor role in the tracking errors in both Parkinsonian and control subjects. It is clear that the technique for determining the visuoperceptual component of performance on complex sensory-motor tasks has considerable scope for application in studies of a variety of brain disorders. PMID- 9214818 TI - Morphological feature extraction for the classification of digital images of cancerous tissues. AB - This paper presents a new method for automatic recognition of cancerous tissues from an image of a microscopic section. Based on the shape and the size analysis of the observed cells, this method provides the physician with nonsubjective numerical values for four criteria of malignancy. This automatic approach is based on mathematical morphology, and more specifically on the use of Geodesy. This technique is used first to remove the background noise from the image and then to operate a segmentation of the nuclei of the cells and an analysis of their shape, their size, and their texture. From the values of the extracted criteria, an automatic classification of the image (cancerous or not) is finally operated. PMID- 9214819 TI - Optimized sampling and parameter estimation for quantification in whole body PET. AB - Whole-body positron emission tomography (PET) has recently emerged as an important imaging tool for cancer detection and staging. Initial applications of the technique have been primarily qualitative. One of the major reasons is the limits imposed by kinetically undersampled data over the whole body, as opposed to the standard method of continuous dynamic sampling in one body location. In this paper, a new estimation method using weighted nonlinear least squares (WNLS) for the first bed position and Bayesian regression (BR) for subsequent positions is proposed. A general criterion for designing optimal sampling schedules which maximizes the measurement information with multiple bed positions is developed. The overall approach is illustrated with the problem of estimating the metabolic rate of glucose (MRGLu) in tumors at different axial positions (image bed positions) in the body by using computer simulations and patient data. The results show that estimates of MRGLu using sparse data and the optimized Bayesian approach are comparable with those obtained by standard methods and fully sampled data. This study demonstrates the potential of the technique described for quantification where several bed positions have to be used to image all the regions of interest (ROI). PMID- 9214821 TI - A 3-D SAR model for current source interstitial hyperthermia. AB - A three-dimensional (3-D) model is presented for the calculation of the specific absorption rate (SAR) in human tissue during current source interstitial hyperthermia. The model is capable of millimeter resolution and can cope with irregular implants in heterogeneous tissue. The SAR distribution is calculated from the electrical potential. The potential distribution is determined by the dielectric properties of the tissue and by the electrode configuration. The dielectric properties and the current injection of the electrodes are represented on a 3-D uniform grid. The calculated potential at an electrode current injection point is not the actual electrode potential at that point. To estimate this potential a grid independent representation of an electrode together with an analytical solution in the neighborhood of the electrode are used. The calculated potential on the electrode surface is used to estimate the electrode impedance. The tissue implementation is validated by comparing calculated distributions with analytical solutions. The electrode implementation is verified by comparing different discretizations of an electrode configuration and by comparing numerically calculated electrode impedances with analytically calculated impedances. PMID- 9214820 TI - An inverse method to optimize heating conditions in RF-capacitive hyperthermia. AB - An inverse method to directly optimize the electrode configuration (positions, sizes, and driving voltages) for radio frequency (RF) capacitive hyperthermia was proposed. The main algorithm, based on the two-dimensional finite element method (2-D-FEM) solution of Laplace and bio-heat transfer equations, iteratively modified the individual boundary potentials around an object thereby making a calculated temperature distribution approach a target temperature distribution. A penalty function governed continuity and smoothness among the boundary potentials so that the optimized boundary potentials became attainable for two plate electrodes. Case simulations demonstrated the viability of the algorithm. For instance, in a computed tomography (CT)-based human abdomen model which had deep and shallow-seated tumors, the optimized electrodes produced a temperature distribution suitable for heating the tumors; the average temperature differences between the tumors and normal tissues were 3.5 degree C for the deep-seated and 7.6 degree C for the shallow-seated tumors within 600 s of heating. A drawback with the present algorithm is that the choice of penalty coefficient and modification of the boundary potentials to coincide with the use of two plate electrodes are carried out manually. These procedures would be automated. PMID- 9214822 TI - Spectral characterization and classification of Carpentier-Edwards heart valves implanted in the aortic position. AB - This paper demonstrates an improvement in the performance of spectral phonocardiography, combined with pattern recognition techniques for monitoring the condition of bioprosthetic heart valves. The analysis of the heart sounds is performed using a modified forward-backward overdetermined Prony's method. Results show that the condition of the bioprosthesis affects mostly the higher part of the spectrum (i.e., above 250 Hz) where no frequency components were found for malfunctioning cases. Therefore, the amplitudes of the three highest frequency components are used as the input vector of an adaptive single layer perceptron-based classifier to identify normal and malfunctioning classes. For the sample set examined, this method gives 100% correct discrimination between normal and malfunctioning Carpentier-Edwards (C-E) valves. PMID- 9214823 TI - Current dipole localization with an ideal magnetometer system. AB - The goal of the study was to explore the most fundamental aspects of a magnetoencephalography (MEG)-based dipole source analysis. For that purpose, a MEG measurement with an ideal magnetometer system (providing the radial component of the magnetic field as a continuous function) is considered. The analytical formulas derived for the variances and covariances of the parameter estimation errors, validated by means of Monte Carlo simulations, allow quantitative predictions in terms of dipole depth, radius and span of the magnetometer system, signal-to-noise (SNR) ratio, and other parameters. A negative correlation exists between radial coordinate and longitudinal component of the moment (perpendicular to radial direction, same plane as actual dipole moment and center of sphere), whereas the other parameters are independent. The standard deviations of the five dipole parameters show fundamental differences with respect to their asymptotic behavior for deep dipoles: If the root mean square (rms) value of the magnetic field is kept constant (moment with depth-dependent amplitude), the error for the transverse coordinate (perpendicular to radial and longitudinal coordinate) is proportional to the distance R between dipole and center of sphere, the errors for the other dipole coordinates, and the relative error for the transverse component of the dipole moment are constant, and the relative error for the longitudinal component of the dipole moment follows a 1/R law. PMID- 9214824 TI - A versatile mechanical ventilator (DIGIT) with high flow stability and a programmable inspiratory phase flow pattern. AB - The paper describes the general characteristics of a newly developed nonconstant flow generator for automatic ventilation of the lungs. It is known that the application of very high pressure to high internal resistance leads to a very stable flow, in that the flow itself is unaffected by external load (patient) variations. The stability of the flow means that the inspiratory process can be controlled by means of the ventilated volume, thus extending DIGIT utilization to high resistance patients. The modulation of the flow is implemented via a digital electromechanical system, which allows the ventilator functions to be accurately programmed. The desired flow waveform is obtained by means of a series of pneumatic valves, the apertures of which are digitally controlled. The design is innovative in that it allows the flow waveform in each of the ten digitalized time steps into which each inspiratory phase is divided to be both programmed and controlled. Other ventilators commercially available and currently in use do not have this functional capability, as they are all designed to model the integral flow of the inspiratory waveform without being able to modify the subunit time steps of a single inspiratory phase. The paper also discusses the results of fundamental tests concerning the performance characteristics of the ventilator. PMID- 9214825 TI - Combined head and eye tracking system for dynamic testing of the vestibular system. AB - We present a combined head-eye tracking system suitable for use with free head movement during natural activities. This system provides an integrated head and eye position measurement while allowing for a large range of head movement (approx 1.8 m of head translation is tolerated). Six degrees of freedom of head motion and two degrees of freedom of eye motion are measured by the system. The system was designed to be useful for the evaluation of the vestibulo-ocular reflex (VOR). The VOR generates compensatory eye movements in order to stabilize gaze during linear or rotational motion of the head. Current clinical and basic research evaluation of the VOR has used a restricted range of head motion, mainly low-frequency, yaw rotation. An integrated eye-head tracking system such as the one presented here allows the VOR response to linear and angular head motion to be studied in a more physiologically relevant manner. Two examples of the utility of the integrated head and eye tracking system in evaluating the vestibular response to linear and angular motion are presented. PMID- 9214826 TI - Dominant frequency analysis of EEG reveals brain's response during injury and recovery. AB - A new method of monitoring an analyzing electroencephalogram (EEG) signals during brain injury is presented. EEG signals are modeled using the autoregressive (AR) technique to obtain the frequencies where there are peaks in the spectrum. The powers at these dominant frequencies are analyzed to reveal the state of brain injury during an experimental study involving progressive hypoxia, asphyxia, and recovery. Neonatal piglets (n = 8) were exposed to a sequence of 30 min of hypoxia (10% oxygen), 5 min of room air, and 7 min of asphyxia. They then received cardiopulmonary resuscitation and were subsequently monitored for 4 h. An optimal AR model order of six was obtained for these data, resulting in three dominant frequencies. These dominant frequencies, referred to as the low, medium, and high frequency components, fell in the bands 1.0-5.5 Hz, 9.0-14.0 Hz, and 18.0-21.0 Hz, respectively. A remarkable feature of our data is the spectral dispersion, or diverging trends in the three frequency bands. During hypoxia, the relative powers of the medium and high-frequency components of EEG increased up to 160% and 176%, from their respective baseline values. During the first minute of asphyxia the medium- and high-frequency powers (relative to baseline) increased by 280-400%. The power in three frequency components went down to nearly zero within 40-80 s of asphyxia. During recovery, the phenomenon of burst suppression was clearly exhibited in the low-frequency component. A new index, called mean normalized separation, representing the degree of disproportionality in the recovery of powers of the three dominant components relative to their mean recovered power, is presented as a possible single indicator of electrical function recovery. In conclusion, dominant frequency analysis helps reveal the brain's graded electrical response to injury and recovery. PMID- 9214828 TI - Feasibility of ultrasound hyperthermia with waveguide interstitial applicator. AB - Interstitial and intracavitary ultrasonic hyperthermia applicators facilitate well-controlled power deposition in tissues. In this paper, analysis of temperature elevation and experimental results in tissue phantom, animal tissue in vivo and animal tissue in vitro are presented for a waveguide applicator intended for treatment of brain tumors. It consisted of a G18 hypodermic needle attached via a conical velocity transformer to a 12.7-mm-diameter piezoelectric disk operated at 1.0 MHz. The axial acoustic pressure distribution had a standing wave pattern with the four cycles/cm spatial periodicity. This periodicity was absent in the temperature distribution in tissue phantoms. The simulations based on a solution to the effective heat conductivity equation indicated that the hyperthermic range can be reached within a 4- and a 10-mm radius around the applicator for a 21- and a 60-mm sample diameters, respectively, with reasonable input power. The first number corresponded closely to the 5-mm radius observed in porcine brain in vivo and the second one came close to the 9-mm radius in porcine brain in vitro. The presented results demonstrate the potential of the ultrasound waveguide interstitial applicator for hyperthermia of small volume tumors. PMID- 9214827 TI - Optimal head related transfer functions for hearing and monaural localization in elevation: a signal processing design perspective. AB - Localization of sound sources by human listeners has been widely studied and theories and various models of the localization and hearing mechanism have been constructed. In the classical "duplex" theory, sound localization in azimuth is explained by interaural time or equivalently, phase differences at low frequencies, and by interaural amplitude differences at higher frequencies. Head related transfer functions (HRTF's) present a linear system approach to modeling localization by representing the direction-dependent transformation the sound undergoes at each ear. Localization in elevation is explained by directional differences in the HRTF's, which also explains monaural localization. We conjecture that the HRTF's evolved during the course of nature (due to the evolution of the shape and structure of the ear etc.) are optimal with respect to several physically realizable criteria. In this paper, we investigate the problem of defining the design constraints which when optimized yield a set of HRTF's for hearing and monaural vertical localization in an attempt to better understand, and if possible, duplicate nature's design. We pursue an engineer's design perspective and formulate a constrained optimization problem, where the desired set of HRTF's is optimized according to a cost function based on several criteria for localization, hearing and smoothness, and also by imposing physically realizable constraints on the HRTF's such as nonnegativity, energy etc. The value of the cost function for a candidate set of HRTF's is an indication of the similarity of that set of HRTF's with respect to the ideal solution (measured HRTF data). The final optimization results we present are similar to the actual HRTF's measured in human subjects, and the associated cost function values are found to be almost equal. This points to the fact that the optimization criteria defined are quite relevant. The significant outcome of this research is the identification of a relevant set of mathematical criteria that could be optimized in the human auditory system to facilitate good hearing and localization. These criteria along with the associated constraints represent the desirable characteristics of the HRTF's in an HRTF-based localization system, and could lead to a better understanding and modeling of the auditory system. PMID- 9214829 TI - Development of an Optical Triplicator for intravital video microscopy of oxygen saturation. AB - The Optical Triplicator produces three copies of a portion of a microscopic image and places them side-by-side on the face of a video image tube, so that all three images can be viewed simultaneously in each video frame. The Optical Triplicator was used in an intravital microscopy assembly to obtain simultaneous images of a microvessel at three visible wavelengths selected to enable the accurate determination of oxygen saturation in microvessels of the hamster retractor muscle. An image processing system was used to obtain light intensity and optical density from video recordings made using the triplicator. Lumenal oxygen saturation profiles were determined using the measured intensity values and a published three wavelength photometric method. PMID- 9214830 TI - A robust sequential detection algorithm for cardiac arrhythmia classification. AB - In [1] qnd [2] Thakor et al. describe a sequential probability ratio test (SPRT) based on threshold crossing intervals (TCI) for the discrimination of ventricular fibrillation (VF) from ventricular tachycardia (VT). However, in applying their algorithm to data from the MIT-BIH malignant arrhythmia database, we observed some overlap in the distributions of TCI for VF and VT resulting in 16% overall error rate for the discrimination. In this communication, we describe a modified SPRT algorithm, using a new feature dubbed blanking variability (BV) as the basis for discrimination. Using the MIT-BIH database, the preliminary results showed that the proposed method decreases the overall error rate to 5%. PMID- 9214831 TI - Magnetometer spacing criterion for biomagnetic source current imaging. AB - A criterion for determining the maximum spacing between magnetometers for measuring the magnetic field is derived. A two-dimensional (2-D) filter model is employed to determine the maximum spatial frequency component present in the magnetic field that is above the spectral noise level. This maximum frequency component is then sampled at a rate greater than twice per period as indicated by the Nyquist criterion, yielding the required magnetometer spacing. It is shown that the rule-of-thumb employed in current clinical biomagnetic array systems, that the spacing between the coils should be approximately equal to the depth of the source, is adequate when the signal-to-noise power ratio is less than 28.4 (14.5 dB). The analysis also quantitatively demonstrates that reducing the separation between the measurement and source planes has a greater effect on the resolution than decreasing the noise level by the same factor. This result is important for employing high Tc superconductor magnetometers that allow thinner thermal insulating layers at the cost of higher thermal noise. PMID- 9214832 TI - Discrete versus syncytial tissue behavior in a model of cardiac stimulation--I: Mathematical formulation. AB - This paper presents a model describing the steady-state response of a two dimensional (2-D) slice of myocardium to extracellular current injection. The model incorporates a continuous representation of the multicellular, syncytial cardiac tissue based on the bidomain model. The classical bidomain model is modified by introducing periodic conductivities to better represent the electrical properties of the intracellular space. Thus, junctional discontinuity between abutting myocytes is reflected in the macroscopic representation of cardiac tissue behavior. Since a solution to the resulting coupled differential equations governing the intracellular and extracellular potentials in the tissue preparation is not computationally tractable when traditional numerical approaches, such as finite element or finite difference methods are used, spectral techniques are employed to reduce the problem to the solution of a set of algebraic equations for the transform of the bidomain potentials. Further, the solution to the "periodic" bidomain problem in the Fourier space is decomposed into two separate solutions: One for the classical-bidomain potentials where it is assumed that the intracellular conductivity values along and across cells incorporate the average contribution from cytoplasm and junction, and another for the junctional potential component. The decomposition of the total solution allows to approximately solve for the junctional component thus achieving high overall computational efficiency. The results of simulation are presented in an accompanying paper. PMID- 9214833 TI - Discrete versus syncytial tissue behavior in a model of cardiac stimulation--II: Results of simulation. AB - The research presented here combines mathematical modeling and computer simulation in developing a new model of the membrane polarization induced in the myocardium by the applied electric field. Employing this new model termed the "period" bidomain model, the steady-state distribution of the transmembrane potential is calculated on a slice of cardiac tissue composed of abutting myocytes and subjected to two point-source extracellular current stimuli. The goal of this study is to examine the relative contribution of cellular discreteness and macroscopic syncytial tissue behavior in the mechanism by which the applied electric field alters the transmembrane potential in cardiac muscle. The results showed the existence of oscillatory changes in the transmembrane potential at cell ends owing to the local resistive inhomogeneities (gap junctions). This low-magnitude sawtooth component in the transmembrane potential is superimposed over large-scale transmembrane potential excursions associated with the syncytial (collective) fiber behavior. The character of the cardiac response to stimulation is determined primarily by the large-scale syncytial tissue behavior. The sawtooth contributes to the overall tissue response only in regions where the large-scale transmembrane potential component is small. PMID- 9214835 TI - A parametric examination of VLSI-based neuronal models of cyclic and reciprocal inhibition. AB - As a prelude to a silicon implementation of a live locomotory network, the phenomena of tonic excitation in single cells, reciprocal inhibition in pairs of cells, and recurrent cyclic inhibition in rings of five cells were recreated and subjected to parametric tests of oscillatory range and stability. Various networks were constructed from comprehensive very large scale integration (VLSI) based artificial neurons and their parametric stability with respect to cellular threshold, refraction, and synaptic weight observed. Circuit tests demonstrated that all three oscillator topologies operated over a broad range of cellular and network frequencies. It was also noted that cells of reciprocal oscillators must possess some measure of short-term synaptic plasticity while those in the cyclically inhibited networks did not. This suggests that the two oscillator types utilize different temporal mechanisms. In parametric tests, tonic cells were sensitive to threshold, refraction, and synaptic weight. Cyclic networks were found to be sensitive to cell threshold, yet less so to refraction. Conversely, the reciprocal circuits were found to be sensitive to refraction, yet less so to cell threshold. This complementary relationship suggests a stability advantage for biological oscillatory networks that incorporate both types. PMID- 9214834 TI - Modeling of mechanical dysfunction in regional stunned myocardium of the left ventricle. AB - Reversible mechanical dysfunction of the myocardium after a single or multiple episode(s) of coronary artery occlusion has been observed in previous studies and is termed myocardial stunning. The hypothesis that stunning could be represented by a decrease in maximum available muscle force in the stunned region was examined by means of a mathematical model that incorporates series viscoelastic elements. A canine experimental model was also employed to demonstrate depressed contractility and a consistent delay of shortening in the stunned region. The mechanical model of the left ventricle was designed to include a normal and stunned region, for which the stunned region was allowed to have variable size. Each region consisted of a volume and time dependent force generator in parallel with a passive elastic force element. The passive elastic element was placed in series with a constant viscosity component and a series elastic component. The model was solved by means of a computer. Passive and active properties of each region could be altered independently. The typical regional measures of muscle performance such as percent shortening, percent bulge, percent thickening, delay of shortening, percent increase in end-diastolic length and other hemodynamic measures were computed. These results were similar to those observed in animal models of stunning. In addition, a nearly linear relationship with end-diastolic length and delay of shortening was predicted by the model. It was concluded that a decrease in the peak isovolumic elastance and augmentation of viscosity effect of creep during stunning can explain mechanical abnormalities of stunned myocardium. PMID- 9214836 TI - Nonstationarity broadening reduction in pulsed Doppler spectrum measurements using time-frequency estimators. AB - The spectral width of Doppler signals is used as measure of lesion-induced flow disturbance. Its estimation accuracy is compromised using the conventional short term Fourier transform (STFT) since this method implicitly assumes signal stationarity during the signal window while the Doppler signals from arteries are markedly nonstationary. The Wigner-Ville (WVD), Choi-Williams (CWD) and Bessel distributions (BD), specifically designed for nonstationary signals, have been optimized for spectral width estimation accuracy and compared to the STFT under different signal to noise ratios using simulated Doppler signals of known time frequency characteristics. The optimum parameter values for each method were determined as a Hanning window duration of 10 ms for the STFT, 40 ms for the WVD and CWD and 20 ms for the BD and dimensionless time-frequency smoothing constant values of five in the CWD and two in the BD. Thresholding was used to reduce the effect of cross terms and side lobes in the WVD and BD. With no added noise the WVD gave the lowest estimation error followed by the CWD. At signal-to-noise ratios (SNR's) of 10 dB and 20 dB the CWD and BD had similar errors and were markedly better than the other estimators. Overall the CWD gave the best performance. PMID- 9214837 TI - Heat transfer evaluation of the nasal thermistor technique. AB - When analyzing transvalvular and venous flow velocity patterns, it is important to relate them to respiration. An accurate recording of respiratory phase can be carried out with different methods. One of these methods is the use of a thermistor, which reacts to the variation in air temperature, placed in the noise of the patient. The thermistor used has a diameter for 1.0 mm and is of standard bead type. Although small, it has a considerable long time-constant and a long time-delay. The high time-constant gives a low cutoff frequency, well below the respiratory frequency and thereby causing a large phase difference. The thermistor was analyzed with the lumped heat capacity method, where it was easy to study the influence from design parameters, time-dependent air temperature, and velocity. The analysis was extended using the finite element method and the temperature field in the thermistor and the probe was calculated as a function of space and time. These calculations confirmed the result from the lumped model. The result showed that timing of respiration was not accurately obtained with the thermistor analyzed. To improve the timing, it was necessary either to change the measuring method or to use signal processing in order to achieve faster response. PMID- 9214838 TI - A method for determining high-resolution activation time delays in unipolar cardiac mapping. AB - This paper presents a method for determining activation time delays in unipolar cardiac mapping data to resolutions considerably smaller than the sample interval. The method involves taking two filtered, differentiated electrograms and computing the Hilbert transform of their cross correlation, which exhibits a negative-to-positive zero crossing at the delay time between the signals. Simultaneous endocardial/epicardial recordings of sinus rhythm were made in the swine right atrium using identical, precisely superpositioned electrode arrays. Data were amplified, lowpass filtered, and digitized at 1000 Hz. A window of data was chosen around each electrogram in an endocardial/epicardial electrogram pair. The windowed electrograms were differentiated and highpass filtered, and the Hilbert transform of the cross correlation between the electrograms was computed. The activation time delay was taken to be the first negative-to-positive zero crossing. Average activation time delays (+/- SD) were computed for 4-s sinus rhythm recordings from each endocardial/epicardial electrode pair. For a representative site, the average transmural activation time delay was 0.71 +/- 0.06 ms (n = 10 electrograms). Time delays estimated using the Hilbert transform method were compared with time delays estimated using the maximum negative slope criterion. The Hilbert transform results exhibited much smaller standard deviations, indicating that the Hilbert transform method may produce more accurate time delay estimates than the maximum negative slope method. PMID- 9214839 TI - Superconducting receiver coils for sodium magnetic resonance imaging. AB - We present the results from sodium magnetic resonance imaging (MRI) experiments using high-temperature superconducting (HTS) receiver coils. Sodium imaging has been shown to have great potential for the assessment of cell integrity but suffers from a substantially lower signal-to-noise ratio (SNR) than that of a hydrogen imaging. The use of an HTS receiver coil was found to significantly increase the SNR relative to an equivalent copper receiver coil at room temperature. The SNR gains afforded by HTS coils can also be used to decrease the imaging time. PMID- 9214840 TI - On measuring curvature and electrical diffusion coefficients in anisotropic myocardium: comments on "effects of bipolar point and line simulation in anisotropic rabbit epicardium: assessment of the critical radius of curvature for longitudinal block". AB - Notion of "curvature" and "propagation perpendicular to the activation front" inherited from electrophysiological theory of isotropic reaction-diffusion media do not apply directly to experimental data taken in uniformly anisotropic myocardium. They apply directly only after the concept of "curvature" is normalized and propagation is made to look perpendicular to activation fronts by rescaling distances to achieve isotropy. Without rescaling, the curvature dependence of longitudinal propagation speed turns out counter-intuitively to measure transverse intercellular electrical coupling. PMID- 9214841 TI - [Regulatory mechanisms for B lymphocyte apoptosis]. PMID- 9214842 TI - [Detection of genetic alterations in human cancer]. PMID- 9214843 TI - [Expression of monocyte chemotactic activity by cross-linked dimerization of a ribosomal protein]. PMID- 9214844 TI - [Respiratory chain of thermophilic bacilli]. PMID- 9214846 TI - [Libraries as tools to explore the specificities of molecular interactions]. PMID- 9214845 TI - [Recent progress in the analysis of the intranuclear organization of the animal cell genome by FISH]. PMID- 9214847 TI - [Streptococcal cytolysins]. PMID- 9214848 TI - The significance of hemispheric localization of ischemic brain lesion for the early prognosis of ischemic brain disease. AB - The possibility of early prediction of ischemic brain disease (IBD) is extremely significant for the creation of the individually designed treatment program. The aim of this study was to determine the significance of hemispheric localization of ischemic brain lesion as possible factor for the early prediction of IBD outcome. The study included 170 patients with IBD, whose score of neurologic and functional impairment was determined after admission by using the standard scales. The values of hemorrheological parameters-erythrocyte sedimentation rate (ESR), fibrinogen and alpha 2 globulin were also determined, so as the extensity of ischemic lesion by CT and MR brain examinations. After the treatment, the score of neurologic and functional impairment was also determined and the degree of ischemic disturbance was estimated-transitory ischemic attack (TIA), reversible ischemic attack (RIA) and cerebral infarction. The lower scores were noticed for neurologic and functional impairment, meaning the more severe degree of ischemic disorder with ischemic lesion localization in right cerebral hemisphere (RCH), in acute stage and at the end of treatment. The higher values of ESR, slightly less of fibrinogen and alpha 2 globulin were revealed in the case of ischemic lesion localization in RCH, so as the higher frequency of RCH lesions in percentages connected with the more severe ischemic disorders. It was concluded that the ischemic lesion localization in RCH might be the predictor of poorer IBD outcome. PMID- 9214849 TI - [Pharmacologic treatment of lipid metabolism disorders]. AB - The aim of the treatment of dyslipidaemia is the primary and secondary prevention of coronary heart disease (CHD). Dietary therapy is the first line in the management of hyperlipidaemia. Lipid-lowering drugs should be used in patients with an inadequate dietary response, with CHD and/or multiple CHD risk factors. The choice of drug depends on the lipid disorder type, the desired plasma lipids reduction and presence of contraindications. Lipid-lowering drugs-anion-exchange resins, nicotinic acid and acipimox, fibrates, statins, probucol and two new classes used in experimental studies (ansamycins and ACAT inhibitors) are presented. Antiatherosclerotic properties of statins are characterized. PMID- 9214850 TI - Conflicting objectives in chemotherapy with drug resistance. AB - A system of differential equations for the control of tumor cells growth in a cycle nonspecific chemotherapy is presented. Spontaneously acquired drug resistance is accounted for, as well as the evolution in time of normal cells. In addition, optimization of conflicting objectives forms the aim of the chemotherapeutic treatment. For general cell growth, some results are given, whereas for the special case of Malthusian (exponential) growth of tumor cells and rather general growth rate for normal cells, the optimal strategy is worked out. The latter, from the clinical standpoint, corresponds to maximum drug concentration throughout the treatment. PMID- 9214851 TI - Gene therapy of T helper cells in HIV infection: mathematical model of the criteria for clinical effect. AB - This paper presents a mathematical analysis of the criteria for gene therapy of T helper cells to have a clinical effect on HIV infection. The analysis indicates that for such a therapy to be successful, it must protect the transduced cells against HIV-induced death. The transduced cells will not survive as a population if the gene therapy only blocks the spread of virus from transduced cells that become infected. The analysis also suggests that the degree of protection against disease-related cell death provided by the gene therapy is more important than the fraction cells that is initially transduced. If only a small fraction of the cells can be transduced, transduction of T helper cells and transduction of haematopoietic progenitor cells will result in the same steady-state level of transduced T helper cells. For gene therapy to be efficient against HIV infection, our analysis suggests that a 100% protection against viral escape must be obtained. The study also suggests that a gene therapy against HIV infection should be designed to give the transduced cells a partial but not necessarily total protection against HIV-induced cell death, and to avoid the production of viral mutants insensitive to the gene therapy. PMID- 9214852 TI - Understanding drug resistance for monotherapy treatment of HIV infection. AB - The purpose of this study was to investigate strategies in the monotherapy treatment of HIV infection in the presence of drug-resistant (mutant) strains. A mathematical system is developed to model resistance in HIV chemotherapy. It includes the key players in the immune response to HIV infection: virus and both uninfected CD4+ and infected CD4+ T-cell populations. We model the latent and progressive stages of the disease, and then introduce monotherapy treatment. The model is a system of differential equations describing the interaction of two distinct classes of HIV--drug-sensitive (wild type) and drug-resistant (mutant)- with lymphocytes in the peripheral blood. We then introduce chemotherapy effects. In the absence of treatment, the model produces the three types of qualitative clinical behavior--an uninfected steady state, an infected steady state (latency), and progression to AIDS. Simulation of treatment is provided for monotherapy, during the progression to AIDS state, in the consideration of resistance effects. Treatment benefit is based on an increase or retention in CD4+ T-cell counts together with a low viral titer. We explore the following treatment approaches: an antiviral drug which reduces viral infectivity that is administered early--when the CD4+ T-cell count is > or = 300/mm3, and the late- when the CD4+ T-cell count is less than 300/mm3. We compare all results with data. When treatment is initiated during the progression to AIDS state, treatment prevents T-cell collapse, but gradually loses effectiveness due to drug resistance. We hypothesize that it is the careful balance of mutant and wild-type HIV strains which provides the greatest prolonged benefit from treatment. This is best achieved when treatment is initiated when the CD4+ T-cell counts are greater than 250/mm3, but less than 400/mm3 in this model (i.e. not too early, not too late). These results are supported by clinical data. The work is novel in that it is the first model to accurately simulate data before, during and after monotherapy treatment. Our model also provides insight into recent clinical results, as well as suggests plausible guidelines for clinical testing in the monotherapy of HIV infection. PMID- 9214853 TI - On reflection and prejudice. PMID- 9214854 TI - Taking neuroleptic medications: a review. AB - Failure to take neuroleptic medications as prescribed often results in relapse and readmission to a psychiatric facility. It is the responsibility of the psychiatric/mental health nurse to ensure that clients have sufficient knowledge and support to enable them to make informed decisions regarding their medication regimen. In addition, the nurse has an important role as client advocate with regard to this treatment option. However, the issues surrounding noncompliance represent a major challenge to psychiatric/mental health nurses. This paper explores these complex issues surrounding the taking of prescribed neuroleptic medication, in particular, the impact of personal factors, knowledge of neuroleptic medications, therapy factors and the helper relationship upon the medication taking behaviour of psychiatric clients is presented in relation to the literature and recent research findings. Finally, the implications of these factors towards the role of the mental health nurse is discussed. PMID- 9214855 TI - The use of behavioural mapping in a study of seclusion. AB - As part of a larger study into the use of seclusion we set out to examine some of the antecedents that led to patients being secluded in one locked psychiatric ward. Behavioural mapping was used to chart the location of incidents that resulted in seclusion. Incidents were observed in several ward areas but most of these occurred in the day room and the dining room. This finding enabled us to question the ward policy of confining patients to these areas to facilitate nursing observation and management. The policy may have contributed to the number of disturbances by limiting personal space for patients. Literature on body buffer zones, crowding and territoriality is used to clarify and interpret the findings. PMID- 9214856 TI - Social science at the crossroads: what direction for mental health nurses? AB - We explore the current and, it would appear, the increasing opposition by some nurse academics to the notion that the traditional scientific methods are suitable for deriving knowledge of essentially social constructions and events. We caution against pursuing an anti-quantitative rhetoric and argue that a rapprochement is possible between contrasting methodologies. To argue for one methodology to the exclusion of all others is both naive and simplistic. All research traditions can be criticized for failing to do justice to social phenomena; it is therefore incumbent upon nurse researchers to be aware of the relative merits of each approach. An example is provided on how different research methodologies can be profitably combined. PMID- 9214857 TI - Adaptation to the mental health setting: the lived experience of comprehensive nurse graduates. AB - The aim of this qualitative descriptive study was to explore the experience of new comprehensive nurse graduates as they adapted to working in the acute psychiatric setting. Interviews were conducted with four participants, focusing on their current work experiences and how the philosophical beliefs and values derived from their educational preparation fit with those they encountered within the practice setting. The data were analysed by noting common experiences, values and meanings and identifying the themes that emerged. The themes were: transition to practice, conflict, contradiction, structural constraints, and the 'reality' of the psychiatric setting. The results of the study confirm the concern that has been voiced by new graduates about the quality and quantity of current orientation programs. Conflicting values and beliefs concerning the nature of mental health/psychiatric nursing also became evident. It appears that the graduates' Comprehensive nursing preparation may have contributed to their feelings of unease as they attempted to fit their own values and beliefs about nursing with those of the acute psychiatric setting. PMID- 9214858 TI - Getting through ethics committees: partnerships between researchers and clinicians working with mentally ill clients. AB - This paper discusses principles of ethics in nursing research with people suffering from mental illness. It offers a set of protocols which have been successful in obtaining approval from ethics committees to conduct research in the area of mental illness-'getting through'. The paper argues that nurse researchers and clinicians can achieve successful outcomes in mental health nursing research by working in partnership. An important aspect of this partnership is a commitment to uphold the rights of consumers of mental health care. PMID- 9214859 TI - The Rozelle Hospital Winter Symposium: clinical supervision in mental health nursing. PMID- 9214860 TI - Success factors for the future survival of rural hospitals. AB - Rural hospitals are among those at greatest risk in the changing world of healthcare delivery. There are, however, those that are continuing to thrive despite the odds. The common thread among these are factors found in select Midwest rural hospitals. PMID- 9214861 TI - Benchmarking for unrelieved pain in a postanesthesia care unit. AB - The problem of unrelieved pain in the PostAnesthesia Care Unit (PACU) is an outcomes management concern. This article discusses the steps taken by the PACU staff to establish a benchmark for unrelieved pain and to evaluate the effect of the Multidisciplinary Pain Management Initiative (MPMI) action plan on the outcomes of length of stay and cost. PMID- 9214862 TI - Effect of a training program on performance of laparoscopic cholecystectomy. PMID- 9214863 TI - Rural telemedicine--the electronic signal flows in both directions. AB - The following article is a descriptive summary of telemedicine evolution in rural Kansas. Multiple electronic applications are used to manage information, time, and distance. Conservation of limited health related resources is emphasized. PMID- 9214864 TI - Influenza immunization cooperative project. AB - During the 1994 flu season, the Missouri Patient Care Review Foundation (MPCRF) began working on a systems-oriented, hospital-focused influenza immunization project for the 1995-96 flu season with four acute care hospitals. The results of the project indicate that a combination of hospital policies, nurse-driven standing order protocols for immunization, and emphasis on beneficiary education resulted in the highest rate of influenza vaccination. PMID- 9214865 TI - Benchmarking emergency medical services/trauma systems funding in the United States. PMID- 9214866 TI - Are gatekeepers necessary? AB - Primary care physicians, acting as gatekeepers, are being used in an attempt to better control rising healthcare costs. However, the success of gatekeepers is difficult to measure because of the lack of data. The purpose of this article is to outline the function of gatekeepers, address the complications in measuring the benefits, and propose methods for further studies. In addition, we hope to foster a sense of responsibility and emphasize the importance of sharing data among organizations involved in the delivery of healthcare. PMID- 9214867 TI - Self-reported cost of illness and health-related quality of life. AB - PURPOSE: To evaluate how well a rapid, self-report of the costs of illness correlates with health-related quality of life. METHODS: A total of 211 patients, participating in a clinical trial of an arthritis medication, completed a Quality of Well-being scale interview and an 18-item self-assessment of healthcare utilization. Subjects completed both these instruments at each of three time points during the trial. RESULTS: Correlations between the measures and across time suggest that patient-reported costs are associated with quality of life and function. CONCLUSIONS: A self-reported cost of illness measure may provide valuable information. With the increase move to assess cost as well as health outcomes, such rapid self-report techniques may prove useful to health services researchers, healthcare system managers, and clinicians. PMID- 9214868 TI - The "success test:" validating the competencies required for healthcare leadership. AB - A leadership competency model was tailored to the culture and strategic direction of BJC Health System by correlating indicators of effective leader behavior with measures of success in the organization. We present the competency model, describe how it was derived, and enumerate ways it will be used to support BJC's transition to an integrated healthcare delivery and financing system. PMID- 9214869 TI - Benchmarking outcomes for diagnosis and AMI: a multidisciplinary hospital performance improvement project. AB - A multihospital process improvement study was designed to evaluate outcomes related to the treatment of Acute Myocardial Infarction (AMI). Time to diagnosis, time to intervention and outcomes of treatment were assessed through a risk adjusted database allowing individual hospital comparisons to benchmark results. PMID- 9214870 TI - Impact of risk adjustment estimators on ranking physician costs for pneumonia. PMID- 9214872 TI - Men in nursing: growing in numbers. PMID- 9214871 TI - Clinical pathway for pneumonia. PMID- 9214873 TI - Warning: gambling can be hazardous to your health. PMID- 9214874 TI - Responding to poor inservice attendance. PMID- 9214875 TI - CPR. Who decides? PMID- 9214877 TI - Compression therapy for chronic venous stasis ulcers. PMID- 9214876 TI - Believing you can do it. PMID- 9214878 TI - [The ethics committee: useful or only fashionable?]. PMID- 9214879 TI - Partners in parenting. PMID- 9214880 TI - Preventing falls in high-risk patients. PMID- 9214881 TI - Recognizing victims of physical and sexual abuse. AB - The continued victimization of children, women, and the elderly remains a problem for health care professionals. Although signs of physical and sexual abuse are easier to diagnose, patterns of neglect in children and the elderly and the terrorizing of women are not so apparent and require a level of detective work on the part of the nurse. Except for physical and sexual assaults perpetrated by strangers, victims of abuse or neglect are often hesitant to reveal the abuse or to cooperate with health care workers because their safety is often not guaranteed. This challenges nurses to be very skilled in detecting abuse and pursuing a course of action that does not further jeopardize the safety of the patient or victim. Combined competencies in physical assessment, interpersonal relationships, and interviewing are important in the assessment phase. Knowledge of protective laws and of community resources that could be immediately triggered to support the patient and family are critical intervention skills. PMID- 9214882 TI - Violence during pregnancy. AB - Abuse of pregnant women is a solvable health problem. The social image of pregnancy as a time of nurturing, love, and caring must be re-examined, because the evidence sadly is present that for many women abuse characterizes their pregnancies. Routine assessment of abuse with a planned intervention for all women is essential to break the cycle of violence and must be standard care for all women regardless of whether the setting is primary or tertiary care. PMID- 9214883 TI - Women who return to abusive relationships: a frustration for the critical care nurse. AB - Nurses caring for abused women become frustrated and concerned when their patients return to the violent relationship. This disconcerted feeling can hinder the care nurses render to these women, subsequently leaving the victim powerless. Understanding this phenomenon can improve patient care, increase the image of women and, ultimately, help banish violence against women. PMID- 9214884 TI - The violence of rape. AB - Although critical care nurses are cognizant of the physiologic trauma sustained in violent crimes, many have little or no experience in conducting sexual assault examinations. Most health care professionals receive little or no formal training in the collection and integrity of forensic evidence. Mishandling of evidence frequently leaves prosecutors powerless to remove sadistic rapists from the streets. Critical care nurses play a crucial role in assessment, treatment, and prevention of physiologic and psychologic trauma caused from sexual assaults. Formulation of new strategies in collaboration with state and hospital protocols will prevent revictimization from inadequate support and interventions. PMID- 9214885 TI - Child abuse. AB - Child abuse is one of the most common major disorders involving children. Nurses who maintain a high index of suspicion; report abuse and neglect to child protection agencies (as required by law); and initiate proper management protocol can protect and preserve the lives and mental health of vulnerable children. By making a commitment to become educated about child abuse, nurses can significantly increase their ability to act as a medical advocate for all children. A nurse's timely and accurate assessment of every pediatric patient is crucial to the early diagnosis and appropriate treatment of these troubled families. PMID- 9214886 TI - Elder mistreatment. AB - Elder mistreatment is a common clinical problem that affects at least 4% of the population over age 65. It occurs in all socioeconomic and racial groups. If our eyes are open and we know the signs, we will find it in our families, in our communities, in our nursing homes, and in our hospitals. All health care professionals should be familiar with the problem of elder mistreatment, adept at recognizing risk factors for mistreatment, and alert to signs of actual mistreatment. If we can recognize at-risk situations, it may be possible to prevent much of the neglect and self-neglect that constitute the bulk of the APS caseload. At the very least, we certainly must be able to treat any cases of mistreatment that cannot or have not been prevented. Part of the ability to provide care for victims of mistreatment depends on clinical expertise, and part depends on active advocacy for the victims. This means advocating for the patient not only with the caregivers and within the hospital system, but also at the community, state, and federal levels. Without this political advocacy, there will be no resources to back up our clinical exercise. Nurses have already done much to develop the field of elder mistreatment and improve the care of victims of abuse, but there is still a lot of work to do. PMID- 9214887 TI - Firearm violence. Impact on the patient and society. AB - Firearm violence has reached an alarming level. The devastation that results from firearms has severe implications for the individual and society. Interventions for patients suffering firearm injuries must begin at the scene and continue through the many phases of illness including emergency care, critical care, intermediate care, rehabilitation, and home care. Nurses are in the prime position to coordinate the many phases of care, with the outcome being to incorporate the victim back into society. Prevention is the key to reducing firearm violence through awareness, education, and legislation. PMID- 9214888 TI - Caught in the crossfire. Children, guns, and trauma: an update. AB - The dramatic increase in violence-related deaths in the last few years demands that violence be recognized as a public health emergency. Even though mortality rates in children have decreased overall and deaths from injuries have also decreased, the rate of violence-related mortality has increased. Violence does not know the limits of cities, age, sex, or race; everyone, everywhere is affected. The recognition of violence as a leading cause of mortality and morbidity in children must become a national priority. By making a commitment to tackle the problem of violence, nurses and other health care professionals can make a significant contribution to society at large. PMID- 9214889 TI - Minimizing the impact of community violence on child witnesses. AB - Child witnesses respond to violent events in two stages: an immediate reaction to the trauma followed by a response to the trauma and grief. The child's stage of development, circumstances surrounding the incident, and reactions of trusted adults affect responses. Secondary prevention measures during the first stage focus on protection and advocacy, while second stage interventions help the child acknowledge and tolerate the realities of the violent event. Child witnesses are at risk for posttraumatic stress disorder and other long-term social, emotional, and developmental problems. Individual characteristics, early life experiences, and protective factors in the environment contribute to children's resilience and ability to survive and grow into healthy adults. PMID- 9214890 TI - Workplace violence. Prevalence, prevention, and first-line interventions. AB - Workplace violence is increasing, but through education, prevention, and man aging escalating crises, critical care nurses can help to minimize the negative consequences of violence. Critical care nurses are particularly prone to acts of aggression and acts of violence due to the stressful environment for them, their families, and their patients. Developing violence prevention skills and interpersonal communication skills, not always highly valued in a critical care environment, are important steps in deterring workplace violence. Although not all incidents of workplace violence can be prevented, recognizing the individual at risk for violence and intervening on the behalf of all involved is a responsibility of every employee including the critical care nurse. PMID- 9214891 TI - The Oklahoma bombing. Lessons learned. AB - The Oklahoma City bombing experience in April of 1995 provided a unique opportunity to test the effectiveness of an existing disaster plan. The critical care nurses at Columbia Presbyterian Hospital learned valuable lessons about managing intense activity, equipment and supplies, staffing resources, and visitor issues. The degree to which the bombing affected the emotional state of personnel was unanticipated, and leaders learned that critical stress management interventions should be included in every emergency preparedness plan. Additionally, recommendations include using runners for communication; assigning specific roles (supplies, staffing, triage); keeping additional staff in reserve for shift relief; ensuring ample hospital staff members are available to coordinate visitors and media; and setting up record systems to preserve continuity. The unique lessons learned as a result of this terrorist attack can be used by other critical care nurses to understand and refine disaster plans. PMID- 9214892 TI - The aftermath of violence. AB - Violence, with the injuries produced and lingering consequences, affects patients, families, and caregivers. Each case is tragic; however, the opportunity for intervention and proactive approaches to education, prevention, and legislative action abound. Most caregivers agree that the social causes of violence, such as breakdown of family and lack of education opportunities, must be addressed to implement long-term solutions. Recommendations for critical care nurses and all caregivers who deal with the impact of violence in their professional and personal lives include Recognize which problems violence imposes on patients, their families and friends, and health care providers. Identify what interventions could assist in dealing with these issues. Be aware of safety issues associated with victims of violence and take immediate measures to maintain a safe environment. Initiate referrals for rehabilitation services at the earliest opportunity. Broaden knowledge of the resources at the institution where the victim is treated and in the victim's community. Trauma coordinators, social workers patient advocates, and volunteers can often provide guidance. Be involved. Prevention, research, professional and public education, and professional, community and legislative coalitions can provide positive direction. Be tolerant. A disabled or mentally impaired man moving slowly through an express grocery line may be a victim of violence. Families, caregivers, and the public at large are all affected by the patient who has been injured by violence. Caring for a victim of violence presents many opportunities to make a difference as a critical care nurse. PMID- 9214893 TI - [Control of respiratory monitoring in the critical patient]. AB - This study of the safety of patients undergoing mechanical ventilation was undertaken to ensure correct ventilation and to find quality-control elements for patients and nursing staff. Differences in mode programming and ventilation parameters and their registry at the change of nursing shifts, and correct programming of the sensitivity and minimum minute volume alarms and maximum airway pressure alarms were examined. In a two-month period, 8 cross-sectional studies were made of G1 prevalence in each of three nursing shifts. The status of the alarms and sensitivity were recorded when incorrect, as well as whether the ventilation mode and parameters coincided with the change of nursing shift. An analysis was made of: FiO2, PEEP, respiratory rate, minute volume, tidal volume, support pressure level, and control pressure level depending on the ventilation mode. Corrective measures were applied for a month. The effectiveness of these measures was evaluated by making 8 new G2 cross-sectional studies of the same type. Two hundred forty-eight G1 and 250 G2 recordings were made. The G2 studies revealed a generalized reduction in errors for all parameters. We concluded that the corrective measures were effective so these indicators were included in the monitoring of risk areas in the unit quality control program. PMID- 9214895 TI - [Informing the families of patients admitted to an intensive care unit]. AB - The admission of a patient to the intensive care unit (ICU) disturbs the family's normal function. Family members are confronted by a new and difficult situation and react with worry and anxiety. They have different needs, particularly for information and assurance. Only effective communication between the family and health-care team helps. The conditions affecting human communication were analyzed, with emphasis on the situations typical of the hospital environment that negatively influence communication. A bibliographic review was made of useful strategies for improving communication with families. To conclude, recommendations are made for facilitating communication, improving the family's well-being, and easing their adjustment to the disease process. PMID- 9214894 TI - [Burn-out of nurses]. AB - The burn-out rate among nurses in three health-care centers in the province of Barcelona, Spain was studied in a sample of 250 professional nurses who completed a three-part questionnaire that addressed: occupational situation, personal situation, and 20 items. Generally speaking, abnormal levels (0.9%) were not found, but 14.4% of the sample presented a moderate state of "burn-out". These results surprised us because nurses often complain of burn-out, but our study of the syndrome by questionnaire revealed that 84.6% judged their state as optimal. PMID- 9214896 TI - [Calculation of the amount of specimen necessary to estimate a proportion and a medium]. PMID- 9214897 TI - Resolution #8. PMID- 9214898 TI - Get ready to vote. PMID- 9214899 TI - Wade King. PMID- 9214900 TI - Caring for an AIDS patient. PMID- 9214901 TI - Life after cancer. PMID- 9214902 TI - The kindness of strangers--how nursing students really learn. PMID- 9214903 TI - First place winner the nurse I admire most. PMID- 9214904 TI - Nursing: a profession with an upbeat future. PMID- 9214906 TI - 1996 publishers directory. Resource books for nursing students. PMID- 9214905 TI - Tips on how to succeed in nursing school. PMID- 9214907 TI - The gift of life. PMID- 9214908 TI - Managed care and the needs of the elderly. PMID- 9214909 TI - The election is over ... now what? PMID- 9214910 TI - Natalie Wipert. PMID- 9214912 TI - A guide to getting the most out of your convention. PMID- 9214913 TI - Community health nursing: a challenging career. PMID- 9214914 TI - Community-based nursing education at Northeastern University. PMID- 9214915 TI - School nursing. PMID- 9214916 TI - Witnessing my first birth. PMID- 9214917 TI - Symbols and their meanings. PMID- 9214918 TI - New Russian immigrants: health problems, practices, and values. AB - As with many ethnic immigrant groups, the Russian immigrants have experienced drastic cultural shock. Even thought there has been tremendous increases in the number of Russians relocating to the United States of America (U.S.A.), very little health care-related information is available about Russian immigrants. This article explores Russian immigrants' themes of potential stress, health problems, and self-care/self-help strategies as reported by a focus group of Russian immigrants in a large U.S.A. southern metropolitan city. PMID- 9214919 TI - Health and illness concepts for cultural competence with Japanese clients. AB - Large numbers of Japanese nationals influx the United States annually. To enable health professionals to better serve this population, selected health and illness concepts are discussed. These include selfcare and risk behaviors, beliefs and practices related to illness and death, and attitudes toward health professionals. Implications for practice are identified. PMID- 9214920 TI - An ethnographic study of illness perceptions and practices of Yemeni-Arabs in Michigan. AB - The purpose of this study was to explore the illness practices of Yemeni-Arab Americans and to generate illness themes based on informant reports. A convenient sample of 30 Yemeni-Arab American women was selected from Dearborn, Michigan. A content analysis of interview data was the basis for data analysis. The Arabic language was used in all the interviews due to enability of the informants to speak English. Thirty-three illness practices were identified by the study informants. Analysis of interview data indicated that informants relied heavily on religious explanations of illness practices. Several cultural themes were deduced from collected data. Among these were belief in an omnipotent deity who is the cause of all that is, confidence in the rational mind of man and empirical knowledge, susceptibility to disease based on gender, reliance and trust in health care providers and desirability of adapting to change. PMID- 9214921 TI - Perceptions of foreign-born male students who selected baccalaureate nursing education in the United States. AB - To explore why foreign-born male students have opted to acquire their basic nursing preparation in the U.S., a survey was conducted in the Fall of 1994 of baccalaureate seniors and juniors. The results revealed the subjects' reasons for entering an American nursing program, why they selected nursing as a career, their overall perceptions of the program, faculty and student experience, and attitudes toward and from their American counterparts. Methodology and recommendations are included. PMID- 9214922 TI - The intravenous nurse specialist's role in the evolving health care environment. Intravenous Nurses Society. AB - The Intravenous Nurses Society contends that intravenous nurse specialists should be involved wherever infusion therapies are delivered. Although the evolving health care environment has led to an increased number of practice settings, the need for quality patient care is still paramount. Intravenous nurse specialists have the skill and expertise to ensure competent practice, as well as the ability to keep costs down. PMID- 9214923 TI - Neonatal intravenous therapy. AB - Intravenous therapy has played an integral role in the advancement of neonatal care during the last 4 decades. Intravenous access is often needed within minutes of delivery for resuscitation and the administration of IV fluids, medications, blood products, and nutrients. Small premature infants, who once would have died, are now being treated with parenteral nutrition and respiratory support for prolonged periods of time. Multiple and prolonged needs for venous access have obligated the use of various IV access devices. Without the placement of these lifesaving lines, many of the sick and premature neonates in today's neonatal intensive care units would die. If they are to reduce the risks for their smallest patients, nurses who place intravenous or arterial lines in neonates should have a clear understanding of neonatal pathophysiology. This article incorporates unique aspects of neonatal care into a review of the selection of an IV access device and site, monitoring of complications, and problem solving. PMID- 9214924 TI - Intradialytic parenteral nutrition therapy for the malnourished hemodialysis patient. AB - Intradialytic parenteral nutrition (IDPN) is a type of parenteral nutrition therapy designed to meet the unique needs of the malnourished patient with end stage renal disease (ESRD) on chronic hemodialysis (HD). It is given during the HD treatment three times a week through the HD machine blood lines. Because of renal failure, the patient with ESRD must receive a kidney transplant (cadaver or living related donor) to preserve his/her life, or choose one of two forms of renal replacement therapy: HD or peritoneal dialysis (PD). This article will focus on the hemodialysis patient and IDPN therapy. PMID- 9214925 TI - Pediatric oncology: past, present, and new modalities of treatment. AB - Although the past two decades have shown a remarkable improvement in the survival rate of children with malignant disorders, cancer is still the primary cause of death from disease in children who are between 1 and 14 years old. This article gives an overview of past, current, and future direction of pediatric cancer treatment modalities. PMID- 9214926 TI - Experience with PICC at a university medical center. AB - A retrospective evaluation of the use of the peripherally inserted central catheters that were inserted by a small, credentialed nursing staff to provide ongoing venous access in the general adult and pediatric patient population at the University of Rochester Medical Center in Rochester, New York. Between 1993 and 1995, 602 catheters were inserted in 775 suitable patients. Correct catheter tip placement in the superior vena cava was achieved in 95% of placements with a catheter dwell time ranging from less than 1 day to 353 days (average, 24 days). Strict adherence to obtaining and monitoring quality assurance kept catheter complications to a minimum. PMID- 9214927 TI - Infusion therapy with milrinone in the home care setting for patients who have advanced heart failure. AB - Advanced heart failure (AHF) is a serious cause of mortality and morbidity in the United States. The treatment of AHF exacts a great financial and manpower toll on the healthcare infrastructure. The quality of life of patients with AHF also is reduced severely. When it is infused at home, Milrinone, which is an intravenous inodilator agent, reduces the number of hospitalizations. Milrinone decreases the overall treatment costs of AHF. Highly skilled home nursing care is required in the treatment of patients. The home care nurse plays a major role in the outcome of a patient with AHF. PMID- 9214928 TI - Pain, role play, and videotape. Pain management staff development in a community hospital. AB - Videotaped role play was an effective staff development strategy used as an initial exercise in a five-part class to update the pain management skills of experienced nurses. It engaged the participants in learning and stimulated discussion and provided concrete feedback of current clinical practices for comparison with the Agency for Health Care Policy and Research pain management guidelines (Acute Pain Management Guideline Panel, 1992). PMID- 9214930 TI - An evaluation of a nurse extern program. AB - Externships are meant to bridge the gap between the educational and practice setting for nursing. This project began with discussions of outcomes for nurse extern programs. The Schutzenhofer Nurse Activity Scale and the Meleis Nurse Self Description Form were used to measure identified outcomes of professional activities and autonomy. Forty-one participants began the study, but 33 completed the pretest and posttest both. Analysis of data showed a positive correlation in Posttest 1 between age and the Meleis Nurse Self-Description Form (NSDF). Dimensions of professional activity changed most in older students. At the end of Phase 2, analysis of data showed significantly greater professional autonomy. However, no significant change in NSDF was seen throughout time. Participants reported increased self confidence. PMID- 9214929 TI - Implementing effective staff education about advance directives. AB - The Patient Self-Determination Act of 1990 guarantees the right to refuse medical or surgical treatment and the right to draft advance directives. This review of the current literature provides those in nursing staff development and inservice education with an overview of advance directives and their implications for nursing education and practice. Possible core subjects for inclusion in planned, purposeful, advance directive education programs are examined, including cultural sensitivity, facilitator skills, interviewing techniques, legal information, patient autonomy, and reasoning and decision making. This review provides a platform for future research. PMID- 9214931 TI - Using an outside consultant for staff development. AB - In this article the author describes the use of a nurse educator/clinical nurse specialist as an outside consultant to provide a program to educate staff nurses about oncology patient care. The continuing education program included a didactic and clinical practicum. The program provided nurses with specialized knowledge about an expanding patient population, improved the level of oncology patient care, and increased the confidence of the physicians and community members in the level of oncology nursing care. PMID- 9214932 TI - An educational model to introduce staff nurses to continuous quality improvement/total quality management concepts. AB - A new healthcare reforms are implemented and as the Joint Commission on Accreditation of Healthcare Organization standards are revised, the demand for continuous quality improvement/total quality management in acute care settings continues to rise. In this article, the authors describe the process used to introduce nursing staff to continuous quality improvement/total quality management. A curriculum committee of staff nurses, nurse educators, and clinical specialists used the continuous quality improvement process to develop a 1-day program that included didactic and experiential activities geared toward assisting the staff nurse to participate actively in total quality management initiatives. PMID- 9214933 TI - Level of literacy in the nurses aide population. Baseline data for nursing staff development. AB - A study of literacy levels of the nurses aide population working in selected acute-care hospitals showed that a literacy deficit existed, described the type and degree of the deficit, and determined its prevalence according to selected demographic factors. Implications for staff development educators in roles of orientation, inservice, and continuing education are discussed for this population in regard to current changes in healthcare delivery patterns. PMID- 9214934 TI - Word games as a cost-effective and innovative inservice method. PMID- 9214935 TI - Tips for the novice grantseeker: implications for staff development specialists. PMID- 9214936 TI - Evaluation of an orientation system for newly employed registered nurses. AB - In this article, the authors discussed the successful evaluation of an orientation system for newly employed registered nurses in a large teaching hospital using the IOP model. This methodology can be successfully applied to any educational program that is consolidated into an organization's goals. Although not well examined, orientation has been reported to be costly (Bethel, 1992; del Bueno, Weeks, Brown-Stewart, 1987). The system presently used at this hospital uses at least 1 week of a nurse educator's time, 3-10 weeks for a newly employed registered nurse, and 3-10 weeks for a preceptor RN. Such an investment of personnel resources mandates examination of the processes and outcomes of the program to ensure newly employed RNs become competent practitioners as efficiently as possible. The use of the IOP model particularly was useful in examining a complex orientation system in a multicentered hospital. Use of this systematic program evaluation separated the overall orientation process into workable components. Tools, such as the algorithm, allowed for easy visualization and comprehension of the process steps. This was indispensable because of the number and scope of people involved in the orientation program. The evaluation process was impartial and focused on the program steps, not on the individuals. Because of this impartiality, people were able to gather and work cohesively to improve the overall program. Use of the IOP model assisted the nurse educators in determining that PBDS was not achieving the goal of identifying individual learning needs. Rather, PBDS was a useful tool in establishing baseline competency of newly employed RNs. The system clearly identified those individuals who had above average knowledge bases and those individuals who had more learning needs. For those with more learning needs, PBDS provides a starting point for planning a structured orientation. Thus, a Phase II PBDS assessment could be used as a more unit-specific assessment to validate whether the RN has achieved the orientation objectives. Although the IOP model is not a strict research methodology, it is appropriate for examination of a program as fluid and ongoing as this. Finally, ongoing run charts or statistical trends will assist the nurse educators in monitoring the quality and effectiveness of the orientation program. PMID- 9214937 TI - ASPAN's home page. PMID- 9214938 TI - Spousal coping during the surgical wait. AB - The purpose of this study was to identify the coping strategies used by a patient's spouse and the perceived effectiveness of these coping strategies while waiting during a loved one's surgery. Forty spouses, male and female (age 25 to 65 years), waiting during a patient's surgery completed a demographic questionnaire and the Jalowiec Coping Scale. Mean scores of the styles and individual strategies were calculated and ranked in descending order. The optimistic coping strategy of "tried to think positively" was used most often by the subjects. The supportant coping strategy of "prayed or put trust in God" was ranked as the most effective strategy. The nurse who instructs the patient regarding their surgery should also prepare the spouse for the surgical wait. Recommending coping strategies to spouses may be a component of stress inoculation. PMID- 9214939 TI - Parent participation in the postanesthesia care unit: fourteen years of progress at one hospital. AB - Parent participation in postanesthesia nursing practice was instituted in 1983 at Boston's Children's Hospital as part of a continuing emphasis on family participation in the care of hospitalized children throughout the hospital. This report describes the evolution and implementation of this practice. The success of this program is supported by data from quality improvement monitors of the comfort levels of PACU patients before and after their parents were invited to the bedside. Quality improvement monitors of parent satisfaction with involvement in their child's care were also conducted. Nursing exemplars illustrate parental and nursing responses to this program. PMID- 9214940 TI - Total spinal anesthesia after an interscalene block. AB - A case study is presented involving a 22-year-old male who developed total spinal anesthesia after interscalene blockade for an arthroscopic procedure of the shoulder. An understanding of the anatomical structures of the brachial plexus, autonomic nervous system function, and side effects of local anesthetics is presented to assist the perianesthetic nurse in assessing and anticipating patient needs and in clinical decision making. PMID- 9214941 TI - Chicken feet: Part one. AB - Propulsion, as defined in Webster's Dictionary, is a giving of forward motion or urging onward. For the last 8 days, however, I have felt propelled into history reversed to a time and place of cruder things-and if I can keep from crying, I would like to tell you of an incredible journey that I was privileged to take. A journey of awareness of how lucky I am to live in the United States of America. A journey of learning respect for a people struggling against odds that I can hardly imagine. A journey of great emotional upheaval for me, from which I am not sure I will ever recover. PMID- 9214942 TI - Fasting before and after ambulatory surgery. AB - During the last decade, questions about the length of time patients were required to fast before elective operations, as well as when they should resume oral fluids after an outpatient operation, became important issues in anesthesia and surgical practice. This review analyzes reasons for the traditional fasting guidelines and presents recent evidence that has caused rethinking in the guidelines. Current recommendations regarding both the presurgical fasting guidelines and fluid intake requirements for discharge after outpatient surgery are outlined. PMID- 9214943 TI - Practical points in the care of patients recovering from a colostomy. AB - The nursing implications for care of colostomy patients is discussed. Colostomy surgery has more than doubled in the last 15 years, making it common to see several patients with colostomies on medical-surgical floors on a daily basis. Because enterostomal nurses are not always available, staff nurses have the responsibility to provide proper care for these patients. PMID- 9214944 TI - Designing research: ex post facto designs. AB - The research design is the overall plan or structure of the study. The goal of a good research design is to insure internal validity and answer the question being asked. The only clear rule in selecting a design is that the question dictates the design. Over the next few issues this column will cover types of research designs and their inherent strengths and weaknesses. This article discusses ex post facto research. PMID- 9214945 TI - Learning to lead the liberated. AB - The historical relationship between employer and employee has changed dramatically in the past few years. Incentives that were once available to motivate staff are no longer a part of the managers' reward "power." Employees recognize that their manager has little control over many things that impact them, and what they are looking for in the way of security and certainty is not being provided by managers. This fundamental shift in relationship has caused those in management to search from new methods to lead their employees. PMID- 9214946 TI - Role of self-efficacy in birth choice. AB - The incidence of vaginal birth after cesarean section is relatively low, although the procedure is of minimal risk to the mother and fetus. The article reports a pilot study designed to investigate whether a relationship exists between the concept of self-efficacy and delivery choice. The Childbirth Self-Efficacy Inventory, a self-administered questionnaire, was completed by 74 pregnant women. The study found that women choosing elective repeat cesarean delivery had lower self-efficacy scores on the instrument. The results suggest the need for further research using assessment of preoutcome, postoutcome, and self-efficacy expectancy scores with specifically designed classes for women who have had a previous cesarean birth. PMID- 9214947 TI - Risk management issues in the perinatal setting. PMID- 9214948 TI - Couples' attitudes toward childbirth participation: relationship to evaluation of labor and delivery. AB - The article reports a correlational, partial replication study investigating the relationship between couples' prenatal attitudes toward childbirth participation and perceptions of their labor and delivery experiences. The Prenatal Attitude toward Childbirth Participation Scale was administered to 119 couples before and after attendance at a childbirth education course. The Labor Delivery Evaluation Scale, the Labor Agency Scale, and the Delivery Agency Scale were administered after delivery. Results showed that childbirth education affected couples' anticipated levels of control during labor and delivery. Anticipated and perceived levels of control were associated, and perceived levels of control correlated with overall evaluations of childbirth experiences. These findings suggest that childbirth educators and health care providers should focus on participants' opportunities to exercise control during childbirth and on identifying emergency situations during which control must be relinquished to the health care team. PMID- 9214949 TI - Protozoan infection in the perinatal period. AB - Protozoan infections represent an area of concern for advanced practice nurses, particularly those working in rural areas or urban environments with refugee populations and those caring for patients with immunodeficiency-related diseases. Some of these infections have major effects on the fetus and neonate yet pose minimal problems to the mother. Protozoan infections are increasing in prevalence because of poor sanitation, overcrowding, increased foreign travel, and high-risk sexual behaviors. There is a need for public education to promote awareness and prevention of such infections. This emerging public health problem has been reported sporadically in the medical and perinatal nursing literature. This paucity of information may be partly due to the difficulty in diagnosing and managing these infections in the perinatal patient. The article discusses the more common infections caused by protozoa, amebae, and sporozoa: trichomoniasis, giardiasis, amebiasis, and toxoplasmosis. PMID- 9214950 TI - Consent and informed consent in perinatal and neonatal settings. AB - It is commonly accepted in American law that an adult of sound mind has the right of self-determination, that is, the right to say what is to be done to his or her own body. This includes the right to accept and refuse treatment for most medical problems or disorders. During the perinatal period, the woman may be asked to submit to procedures and medications for herself and her fetus/ neonate. Nurses must be sure that patients or parents have given valid consent to any type of touching done in the course of nursing care. Furthermore, nurses must be certain that women have given informed consent to the certified nurse-midwife, nurse practitioner, or physician for any treatments or procedures that have inherent risks. The article defines and describes the concepts of consent and informed consent and examines the roles and responsibilities of perinatal nurses in these processes. PMID- 9214951 TI - Catheter-related sepsis in the neonate: thinking critically about a persistent problem. AB - Catheter-related sepsis (CRS) is a clinical problem gaining increasing attention in the literature. Before strategies for reducing the incidence of CRS can be identified, a consistent definition of the problem must be developed. Critical thinking is an intellectual process used to reflect upon a topic of interest. The critical thinking process provides a framework for comprehensive analysis of patient care problems. The article describes the critical thinking process and proposes a model of critical thinking for practicing nurses. This model is applied to the development of comprehensive definitions for CRS that may be used by neonatal nurses for clinical and research purposes. PMID- 9214952 TI - Neonatal nurse practitioner role satisfaction. AB - The article reports a descriptive study utilizing a triangulated approach to explore the role satisfaction of a nationwide random sample of 315 neonatal nurse practitioners (NNPs). Role satisfaction was found to be high, and practice philosophy tended toward a medical orientation. Those NNPs who were master's prepared and who reported to and were evaluated and paid by departments of nursing were more likely to have a nursing practice philosophy. Intrinisic factors related to satisfaction included autonomy, patient management, relationships with colleagues and families, role diversity, and a sense of accomplishment. Implications for administrators and NNPs are discussed. PMID- 9214953 TI - Adolescent mothers' perceptions of the neonatal intensive care unit environment. AB - The neonatal intensive care unit (NICU) environment has been found to be a major source of distress for older parents, but what about adolescent mothers? A prospective, descriptive study was conducted to describe adolescent mothers' perceptions of the stressors found in the NICU environment. Data were collected using the NICU Parental Stress Scale and a demographic data form. These 46 mothers found that the most stressful aspects of the NICU were parental role alterations and the infant's appearance and behavior. Less stressful were the sights and sounds of the NICU and communication with staff. Nurses should continue to identify and alleviate stressors that can compromise the parenting experience of mothers of all age groups. PMID- 9214954 TI - Advanced practice nursing in the care of the high-risk infant. PMID- 9214955 TI - Electronic fetal monitoring then and now. PMID- 9214956 TI - Family-centered care: controversies and complexities. PMID- 9214957 TI - Pregnancy-induced hypertension, preeclampsia, and eclampsia. PMID- 9214958 TI - Preterm birth in the United States: current issues and future perspectives. PMID- 9214959 TI - The promise of clinical pathways. PMID- 9214960 TI - The advanced practice movement in nursing: impact on perinatal care. PMID- 9214961 TI - The state of perinatal nursing: current and future profiles as described by perinatal nursing service directors. AB - The health care industry is in the throes of remarkably penetrating and destabilizing change, the effects of which have been felt earliest by perinatal service directors. In anticipation of future trends marked by rampant change, a survey of perinatal service directors and vice presidents was conducted to elicit their opinions about the current and future states of perinatal health care. Findings supported the notions that change is a constant, that clinical and service excellence is a mandate, and that collaboration is key. Future success will require many old behaviors and systems to be replaced. Leadership to guide us to the future has never been more important. PMID- 9214962 TI - Twenty-five years of obstetric patient satisfaction in North America: a review of the literature. AB - The North American literature on obstetric patient satisfaction of the past 25 years was reviewed using two major computerized databases. The articles identified by these searches were supplemented with other research articles identified in reference lists. The review highlights the difficulties inherent in the use of many different methodologies to study obstetric patient satisfaction. The main methodologies have been mailed questionnaires, telephone interviews, and semistructured interviews, with data collection periods ranging from 24 hours to 2 years postpartum. The various approaches to data collection make comparison of results among studies exceedingly difficult. The reluctance of patients to criticize their caregivers has been problematic and is evidenced by satisfaction ratings that are positively skewed. Factors that have been reported to be most influential in obstetric patient satisfaction include communication, control, participation in decision making, presence of a support person, information/prenatal classes, nursing care services, length of stay, and physical environment. The relative importance of these factors, however, has not been ascertained. PMID- 9214963 TI - Providing developmentally supportive care in the newborn intensive care unit: an evolving challenge. AB - Clearly, the great majority of premature infants currently survive their neonatal course in the hospital. The major challenge facing neonatal health care providers is to combine the necessary technologic intensive care for the newborn with a sensitive and individualized approach to facilitate neurobehavioral development while acknowledging and supporting the parents of these infants in their role as primary advocates and long-term caregivers. This challenge has been met in a variety of ways over time, and a new history of providing developmentally supportive care is being created. A multidisciplinary collaborative approach based on the synactive theory of newborn development incorporates both environmental and caregiving modifications to enhance infant and family outcomes. The article reviews the history of this approach to care, the current state of the art and practice, and the current challenges for future implementation. PMID- 9214964 TI - Preterm infants and STRESS: a tool for the neonatal nurse. AB - Preterm infants are not physiologically or developmentally prepared for life outside the supportive environment of the mother's womb. Their response to stimuli is often immature and disorganized rather than adaptive. The Roy Adaptation Model's theory of an adaptive person may provide a framework for nurses to assess, plan, and evaluate nursing care for fragile preterm infants. The article examines actual and potential stressors of the premature infant; describes commonly observed disorganized, ineffective responses; and proposes a clinical tool (the STRESS tool: signs of stress, touch interventions, reduction of pain, environmental considerations, state, and stability) that nurses can use when caring for medically fragile infants. PMID- 9214965 TI - Meeting the special nutritional needs of sick infants with a percutaneous central venous catheter quality assurance program. AB - Adequate nutrition is important in infancy because it can affect brain growth. A critical period for brain growth is the end of the fetal growth period and the first 2 years of life. Delivery of nutrition to infants in the neonatal intensive care unit is challenging because illness and prematurity increase nutritional need and create access difficulties. One technique, percutaneous central venous catheter (PCVC) placement, eliminates access difficulties. Nevertheless, safe delivery of parenteral nutrition through PCVCs is dependent on minimizing infectious and mechanical complications. With the implementation of a PCVC quality assurance program, problems can be identified early, and appropriate, timely interventions can be initiated. PMID- 9214967 TI - Bladder control. PMID- 9214966 TI - The rewards of exposing fraud against the government. PMID- 9214968 TI - Alzheimer's disease update. PMID- 9214969 TI - Polio vaccine: what you should know. PMID- 9214970 TI - Nurses: act now! AB - Drawing from the tradition of the theater, we can create a new paradigm for creating humor, health and happiness. The basis of this paradigm: we become what we do. In other words, we can become healthier and more successful by consciously modeling (acting) the happy, healthy traits we desire. In addition, staging, scripting, costuming and acting the part can stimulate our bodies to produce neuropeptides such as endorphins and other chemicals that bolster the immune system and promote well being. PMID- 9214971 TI - My rotation through ER. PMID- 9214973 TI - Americans doubt accuracy of cancer diagnosis. PMID- 9214972 TI - Steroid induced osteoporosis guidelines overview supplied by the American College of Rheumatology. PMID- 9214974 TI - Asthma update. PMID- 9214975 TI - Satisfaction guaranteed. PMID- 9214976 TI - Six in one hand ... dietary value of eggs versus cholesterol risks. PMID- 9214977 TI - All I want for Christmas is... PMID- 9214978 TI - Care of morbidly obese people with spinal cord injury. AB - As the number of obese people in the United States continues to increase, all nurses will encounter the unique challenges that result from the pathophysiological changes in this population. The combination of morbid obesity and spinal cord injury creates complex medical management for caregivers throughout hospitalization and after dismissal. Thus, the complications of spinal cord injury are intensified with obesity. Prevention and treatment of secondary complications require nursing practice to go above and beyond the standards of care. This article reviews complications and adjustments in the care of the morbidly obese spinal cord-injured patient, beginning at the scene of injury and through the emergency trauma, intensive care, and rehabilitation units. PMID- 9214979 TI - The trauma top 10. The top 10 things to know, appreciate, and think about when caring for pediatric trauma patients. PMID- 9214980 TI - [Kidney replacement therapy]. PMID- 9214981 TI - [Chronic kidney failure--etiology and sequelae]. PMID- 9214982 TI - [Child abuse and neglect. Prevention and therapeutic intervention]. PMID- 9214983 TI - [Language development disorders in infants]. PMID- 9214984 TI - [Oral abnormalities in the newborn]. PMID- 9214985 TI - [Anemias in children]. PMID- 9214987 TI - [Travel vaccinations]. PMID- 9214986 TI - [More and less toxic plants]. PMID- 9214988 TI - [Alternative therapies]. PMID- 9214989 TI - [Perspectives in hospital financing]. PMID- 9214990 TI - [Resurrection from the test tube?]. PMID- 9214991 TI - [Danger with and without latex]. PMID- 9214992 TI - [Viewpoints on hospital hygiene--legal aspects]. PMID- 9214993 TI - [Ambulatory care in oncology--the nursing situation in the home]. PMID- 9214994 TI - [Reinforcement of the key position of middle management within the framework of courses in administration]. PMID- 9214995 TI - [Did you find the right method? II. Rotation therapy ought to be financially possible]. PMID- 9214996 TI - [Meanings of illness. From escape to existential change]. PMID- 9214997 TI - [Overcoming depressions. Advice for patients]. PMID- 9214998 TI - [The endoscopic break-up of uretheral calculi with Alexandrite lasers]. PMID- 9214999 TI - Information is critical to the success of the present health care delivery systems. PMID- 9215000 TI - The "rite of passage" for nurse executives of the 21st century is learning the informatics language. PMID- 9215001 TI - Informatics nurse specialist: roles in health care organizations. AB - The cost of managing and processing health care information is a significant component of hospital operating budgets, yet health care lags far behind other industries in the effective use of information technology. As nurses are the largest group of users of health care information, improving nurses' information management capability could have a significant impact. Informatics nurse specialists understand the concepts and technology of nursing information management and can provide operational and strategic benefits to nursing organizations. In this article, educational preparation, certification, and roles of informatics nurse specialists are discussed along with functions of health care information technology and nursing informatics. PMID- 9215002 TI - Intellectual property issues in the digital health care world. AB - The digitization of health care information presents a troublesome challenge to the two core principles of copyright: the expression of an idea should be rewarded and protected for a certain period and the user should be allowed the right to fair quotation during that period. The debate about intellectual property in a digital world will be a major focus over the next few years. Nursing administrators need to become copyright savvy to voice their concerns, protect nursing's works, and ensure that health care information is accessible for the public. PMID- 9215003 TI - Privacy, confidentiality, and security in clinical information systems: dilemmas and opportunities for the nurse executive. AB - Protecting the patient medical record in an information age is neither simple or easy. Confidentiality dilemmas create opportunities for nurse executives to exercise leadership in areas of critical concern. The drive toward computerization offers profound implications for privacy, security, and confidentiality, which must be proactively pursued through the careful development of preventive measures. Additionally, the benefits of clinical information systems (CIS) in patient care must not be minimized. The nurse executive who understands the benefit of CIS is in a position to advocate for its important role in health care today. PMID- 9215004 TI - LDS hospital, a facility of Intermountain Health Care, Salt Lake City, Utah. AB - On-line documentation by nurses and a comprehensive text management system are functional in several facilities of intermountain Health Care (IHC). The following articles detail factors in the design and implementation of this computerized network as experienced at LDS Hospital, part of the IHC system. Areas discussed are the system's applications for medical decision support, communication, patient classification, nurse staffing versus cost, emergency department usage, patient problem/event recording, clinical outcomes, and text publication. Users express satisfaction with the time saving, consistency of reporting, and cohesiveness of these applications. PMID- 9215005 TI - Health status measurement in computer-based patient record systems. AB - As requests for information about quality of health care have increased, purchasers of computer-based patient record (CPR) systems are demanding information regarding the manner in which these systems can assist in measuring health care quality. The type of information requested by purchasers of health care, accrediting agencies, and consumer groups is shifting toward more complex measures of health status. Both provider and patient perceptions of health status are relevant in measuring the impact of health care interventions. Standardized coding and classification systems and standardized health status measurement instruments each offer utility in capturing and representing health status in CPR systems. PMID- 9215006 TI - Value-adding information: virtual conferencing, a telecommunication pathway to the future. AB - The information or digital age has turned information communication upside down and inside out. This article describes use of the Internet and World Wide Web information flow and communication for a virtual professional health national event, in Australia. The stored information and subsequent dialogue were available nationally and internationally, making the telecommunication conference a global one. Defined are the background, vision, and assumptions on which the conceptual conference structure was ultimately designed, implemented, and evaluated, including the implications for value-adding nursing information. PMID- 9215007 TI - Clinical information system: a "gateway" to the 21st century. AB - This article describes the selection, design, and implementation of a clinical information system that includes order entry, results reporting, physical assessments, and critical care. The benefits of implementing this data management system such as cost savings, chart-as-you-go documentation, use of wireless technology, online quality improvement audits, and rapid data retrieval will be explained. PMID- 9215008 TI - The Virginia Henderson International Nursing Library: resource for nurse administrators. AB - This article describes the major knowledge resource of the Virginia Henderson International Nursing Library, The Registry of Nursing. The first part of this article examines informatics issues and is accompanied by examples of retrieval from a typical bibliographic database and a retrieval from the Registry of Nursing Research using case mix, both as a subject heading and as a research variable. The second part of the article examines the interaction of informatics and technology used in the Registry and presents some other Library resources. PMID- 9215009 TI - Nursing data sets are finally beginning to catch up to technology's promise. PMID- 9215010 TI - The expanding role of simulators in risk management. PMID- 9215011 TI - Meningitis after spinal anaesthesia. PMID- 9215012 TI - Inaccurate reporting of simulated critical anaesthetic incidents. AB - Eleven anaesthetists completed a simulated anaesthetic which was deliberately complicated by a slow progressive bradycardia followed by an episode of severe bronchospasm. After the simulation, each anaesthetist was asked to complete an anaesthetic chart and a critical incident report. Considerable discrepancies were found between the anaesthetists' written accounts, a videotape of their performance and actual data from the simulator. During the simulations, all of the anaesthetists reacted appropriately and treated their "patient" successfully but their written accounts showed a tendency to record "typical" rather than actual events and to ignore events not consistent with their final diagnosis. Only four anaesthetists mentioned bradycardia in their written description and none accurately described the changes in arterial pressure during the episode of bronchospasm. The findings are in keeping with other studies which suggest that people record events as "schemata" rather than as collections of discrete facts. These results have significant implications for those involved in the teaching of anaesthesia and in the analysis of critical incidents. PMID- 9215013 TI - Propofol and bradycardia: causation, frequency and severity. AB - As part of the development of a model for the study of adverse events, we have investigated the risk of bradycardia with propofol. A systematic search for any type of report, published and unpublished, was made to review the evidence that propofol increases the risk of bradycardia, asystole and death from bradycardic events. Quantitative and qualitative analyses of data with different strengths of evidence were performed. Sixty-five published and 187 spontaneous reports to drug monitoring centres described with different strength of evidence a biological basis for propofol-induced bradycardia, 1444 bradycardias, 86 asystoles and 24 deaths. In controlled clinical trials, propofol significantly increased the risk of bradycardia compared with other anaesthetics (number-needed-to-harm 11.3 (95% confidence interval 7.7-21)). In paediatric strabismus surgery the number-needed to-harm was 4.1 (3-6.7). One of 660 patients undergoing propofol anaesthesia had an asystole. The risk of bradycardia-related death during propofol anaesthesia was estimated to be 1.4 in 100,000. Data from the phase IV study of propofol did not agree with data from controlled studies. Propofol carries a finite risk for brady-cardia with potential for major harm. Study of adverse events should be made with systematically searched data and, in contrast with study of efficacy, not restricted to randomized, controlled trials. PMID- 9215014 TI - Phantom pain and sensation among British veteran amputees. AB - Using a mail-delivered questionnaire, we surveyed 590 veteran amputees concerning phantom pain, phantom sensation and stump pain. They were selected randomly from a population of 2974 veterans with long-standing limb amputation(s) using a computer random number generator. Eighty-nine percent responded and of these, 55% reported phantom limb pain and 56% stump pain. There was a strong correlation between phantom pain and phantom sensation. The intensity of phantom sensation was a significant predictor for the time course of phantom pain. In only 3% of phantom limb pain sufferers did the condition become worse. One hundred and forty nine amputees reporting phantom pain discussed their pain with their family doctors; 49 were told that there was no treatment available. Transcutaneous electric nerve stimulation, analgesics and non-steroidal anti-inflammatory drugs were satisfactory methods for controlling phantom limb pain. PMID- 9215015 TI - Low molecular weight heparin associated with spinal anaesthesia and gradual compression stockings in total hip replacement surgery. Arar Study Group. AB - The benefit/risk ratio of administering heparin during spinal anaesthesia in patients undergoing total hip replacement (THR) has not been studied widely. We conducted a prospective, randomized, double-blind study to compare low molecular weight heparin (LMWH) for 10 days and placebo in patients undergoing THR performed under spinal anaesthesia associated with gradual compression stockings. Efficacy was assessed by systematic bilateral ascending venography on day 10 +/- 2 in a sequential analysis. Among the 170 patients enrolled, data were available in 153 patients. In the LMWH group (n = 78) the total incidence of deep vein thrombosis (DVT) was 14.1% compared with 37.3% in the placebo group (n = 75) (P = 0.0016). No gross neurological sequelae were observed during the study. This study showed that the addition of LMWH in patients undergoing THR under spinal anaesthesia and wearing gradual compression stockings significantly decreased the incidence of venogram-proved DVT. PMID- 9215016 TI - Intrathecal infusion of bupivacaine with or without morphine for postoperative analgesia after hip and knee arthroplasty. AB - Postoperative pain after major orthopaedic operations can be controlled by continuous intrathecal administration of opioids or local anaesthetics. Effective intrathecal analgesia can be achieved through synergism of low doses of the two analgesic drugs and, possibly, less drug-related adverse effects. Therefore, we have evaluated the usefulness of a combined low-dose bupivacaine and morphine infusion in patients undergoing hip and knee arthroplasty. Spinal anaesthesia was induced in 55 ASA I-III patients with 0.5% bupivacaine 2 ml via a 28-gauge spinal catheter (L3-4 interspace) and 0.5-ml increments were given if needed. Intrathecal 24-h infusions consisted of bupivacaine 2 mg h-1 alone (n = 18), bupivacaine 1 mg h-1 alone (n = 18) or bupivacaine 1 mg h-1 combined with morphine 8 micrograms h-1 (n = 19). The interview after 3, 6, 12 and 24 h included assessment of pain at rest and on movement (VAS scale), occurrence of sensory and motor block and nausea/vomiting. Bupivacaine 1 mg h-1 combined with an infusion of morphine provided as good postoperative analgesia as bupivacaine 2 mg h-1, but motor block disappeared earlier (P = 0.01). Patients in the bupivacaine 1-mg h-1 group required more supplementary doses of oxycodone i.m. than the other groups (P = 0.04). Time to first oxycodone dose from the start of intrathecal infusion did not differ between groups. The frequency of nausea and vomiting was similar in all groups. In spite of this, antiemetic medication was required more often in the bupivacaine 1-mg h-1 group (possible because of opioid rescue medication). On the ward, one patient in the bupivacaine 2-mg h-1 group experienced a new increase in sensory block with concomitant hypotension. One patient in the same group had minor decubitus on the heel of the operated leg, probably because of prolonged motor block. We conclude that intrathecal infusion of a combination of bupivacaine 1 mg h-1 and morphine 8 micrograms h-1 produced adequate postoperative analgesia. Unfortunately, postoperative nausea and vomiting was a frequent disturbing adverse effect. PMID- 9215017 TI - Hyaluronidase and peribulbar block. AB - We have assessed the effect of two concentrations of hyaluronidase on the quality of peribulbar block, using a low volume, single injection technique. We studied 200 patients undergoing elective intraocular surgery, allocated randomly to one of three groups. Group 1 (n = 50) received peribulbar block with 5 ml of a 1:1 mixture of 0.5% plain bupivacaine and 2% plain lignocaine. Group 2 (n = 75) received this solution supplemented with hyaluronidase 50 iu ml-1. Group 3 (n = 75) received the same solution supplemented with hyaluronidase 300 iu ml-1. Lack of ocular motility was considered to be the only objective sign of successful block and movement of each rectus muscle was scored at 1-, 5- and 10-min intervals. If the block was successful at 5 min, the 10-min score was omitted. If the block was unsuccessful at 5 min, a second injection of 2% lignocaine 3 ml was given and additional assessments performed at 5-min intervals. At 1 min, ocular motility scores were significantly lower in group 3 compared with the control group (P < 0.05). The incidence of satisfactory block at 5 min was increased in both groups given hyaluronidase (group 2, P < 0.05; group 3, P < 0.001). There were no significant differences between groups 2 and 3 with respect to quality of block at 5 min. Hyaluronidase in both concentrations improved the quality of peribulbar block at 5 min, and when used in a concentration of 300 iu ml-1, also improved the speed of onset of block. PMID- 9215018 TI - Intraperitoneal lignocaine for pain relief after total abdominal hysterectomy. AB - In this preliminary randomized study, we have measured pain scores at rest and on movement, 24 and 48 h after operation in 19 control patients, who received 50 ml of saline i.p., and in 20 test patients, in whom 50 ml of saline solution containing lignocaine 200 mg and adrenaline 1:500,000 were instilled into the peritoneal cavity after total abdominal hysterectomy. We found that there was no difference in linear analogue scores for nausea, pain on movement or morphine consumption after operation between the two groups, but pain scores at rest were significantly lower in the lignocaine group at 24 and 48 h compared with the saline group. In the lignocaine group, blood sampling over a 3-h period revealed a mean maximum serum concentration of 0.4 microgram ml-1 at 3 h and a highest concentration in any patient of 0.87 microgram ml-1. PMID- 9215019 TI - Pre-anaesthetic assessment of coagulation abnormalities in obstetric patients: usefulness, timing and clinical implications. AB - The usefulness and optimal timing of laboratory coagulation tests before obstetric extradural analgesia are controversial. Moreover, the significance of mild coagulation abnormalities during pregnancy remains unclear. We have assessed the reliability of coagulation tests performed several weeks before delivery as predictors of coagulation abnormalities during labour. Platelet count, plasma fibrinogen concentration, prothrombin time (PT) and activated partial thromboplastin time (aPTT) were sampled in 797 women during the ninth month of pregnancy and checked during labour. Platelet count was less than 100 x 10(9) litre-1 for 11 women during labour. Only three had been detected by the first sample. Platelet count less than 100 x 10(9) litre-1 or fibrinogen concentration less than 2.9 g litre-1 during labour were associated with an increase in the incidence of postpartum haemorrhage (odds ratio = 19.7). We conclude that a platelet count several weeks before delivery was not reliable in predicting thrombocytopenia during labour and that women with mild coagulation abnormalities in early labour may need special attention regarding the risk of postpartum haemorrhage. PMID- 9215020 TI - Effect of progressive haemodilution with hydroxyethyl starch, gelatin and albumin on blood coagulation. AB - We have compared the effects of progressive (30% and 60%) in vitro haemodilution with hydroxyethyl starch (HES), gelatin (GEL) and albumin (ALB) with haemodilution using 0.9% saline in 96 patients by thrombelastography. Haemodilution with HES, GEL and ALB significantly (P < 0.05) compromised coagulation time (k), angle alpha and maximal amplitude (MA), with HES having the most negative effect at 30% and 60% haemodilution (P < 0.05). Haemodilution with saline significantly affected all variables of blood coagulation and clot lysis measured by thrombelastography, resulting in an increased coagulability at 30% haemodilution. To specifically assess the intrinsic effect of plasma expander molecules on blood coagulation and clot lysis, we analysed the difference between saline diluted blood (same degree of haemodilution) and plasma expander diluted blood. Prolongation of reaction time (r) was found for HES at 30% and 60% haemodilution and for ALB at 60% haemodilution and an increase in clot lysis by HES, GEL and ALB became evident. We conclude that HES, GEL and ALB compromised blood coagulation, while the maximum effect was found with HES. PMID- 9215021 TI - Pharmacokinetics and pharmacokinetic-dynamic modelling of rocuronium in infants and children. AB - We have determined the pharmacokinetics and pharmacokinetic-pharmacodynamic relationship of rocuronium in infants and children. We studied infants (n = 5, 0.1-0.8 yr) and children (n = 5, 2.3-8 yr), ASA II, in the ICU while undergoing artificial ventilation under i.v. anaesthesia with an arterial cannula in situ and the EMG of the adductor pollicis muscle was monitored. Rocuronium 0.06 (infants) and 0.09 (children) mg kg-1 min-1 was given i.v. over +/- 5 min until 85% neuromuscular block was obtained. Arterial blood samples were obtained over 240 min. Plasma concentrations were measured by HPLC. Pharmacokinetic-dynamic variables were calculated using the Sheiner model and the Hill equation. Statistical analysis was performed using the Mann-Whitney U test (P < 0.05). The mean administered dose was 0.32 (SD 0.08) mg kg-1 and 0.4 (0.1) mg kg-1 for infants and children, respectively. Infants differed from children in plasma clearance (4.2 (0.4) vs 6.7 (1.1) ml min-1 kg-1), distribution volume at steady state (231 (32) vs 165 (44) ml kg-1), mean residence time (56 (10) vs 26 (9) min), concentration in the effect compartment at 50% block (1.2 (0.4) vs 1.7 (0.4) mg litre-1) and the slope of the concentration-effect relationship (5.7 (1.3) vs 3.9 (0.5)). Calculated mean ED90 values were 0.26 and 0.34 mg kg-1 for infants and children, respectively. The time course of neuromuscular block after equipotent doses did not differ. PMID- 9215022 TI - Use of inhaled nitric oxide in British intensive therapy units. AB - The use of inhaled nitric oxide in the critically ill has increased significantly over the past few years but little published information exists on standards for current practice. Sixty-four intensive therapy units in the UK were surveyed by questionnaire from which 54 (84.4%) satisfactory replies were received. We present the survey results and put forward recommendations based on current literature and our own clinical experience for the safe use of inhaled nitric oxide. PMID- 9215023 TI - Comparison of the effects of ketamine-midazolam with those of fentanyl-midazolam on cortical somatosensory evoked potentials during major spine surgery. AB - Cortical somatosensory evoked potentials (CSEP) allow monitoring of spinal cord function during surgery. Ketamine has been shown to enhance CSEP amplitude, but there is no previous study comparing its effects with those of other anaesthetic regimens. Therefore, we have compared the effects of ketamine with those of fentanyl, both combined with midazolam, on CSEP monitoring during major spine surgery. Twenty patients with normal preoperative CSEP were allocated randomly to a ketamine or fentanyl group. Anaesthesia was induced with ketamine 3 mg kg-1 or fentanyl 6 micrograms kg-1 i.v., and midazolam 0.3 mg kg-1 i.v in both groups, and maintained with continuous i.v infusion of ketamine 2 mg kg-1 h-1 or fentanyl 3 micrograms kg-1 h-1, combined in both groups with midazolam 0.15 mg kg-1 h-1 and 60% nitrous oxide in oxygen. CSEP were elicited by tibial posterior nerve stimulation and measured P1 and N1 latencies, and P1-N1 amplitude, CSEP were recorded before and after induction, at 15 min, 1 and 2 h after induction, during skin closure and after removal of nitrous oxide. Both groups were comparable in characteristics, duration of surgery, mean arterial pressure and temperature. CSEP latencies were not significantly affected in either group. CSEP amplitude decreased significantly over time in the fentanyl group (from mean 2.02 (SEM 0.41) to 0.95 (0.17) microV, P < 0.05), but not in the ketamine group (from 1.33 (0.36) to 1.05 (0.31) microV, ns). Nevertheless, we did not observe any significant differences in amplitudes or latencies between the two groups. The delay in obtaining the first voluntary postoperative motor response was significantly greater in the ketamine group (170 (54) vs 55 (17) min, P < 0.01). Both ketamine and fentanyl allowed us to obtain reliable CSEP during major spine surgery, and there were no significant difference between these two anaesthetic regimens for CSEP monitoring, but a longer delay for voluntary postoperative motor assessment was observed in the ketamine group. PMID- 9215025 TI - Effect of exogenous nitric oxide and superoxide on interleukin-8 from human polymorphonuclear leucocytes. AB - Patients with acute inflammatory lung injury are commonly treated with inhaled nitric oxide. Nitric oxide has profound immunoregulatory effects. Increased concentrations of the cytokine interleukin-8 (IL-8) in bronchoalveolar lavage fluid has been associated with disease severity. We have investigated the effects of a nitric oxide donor and a combined nitric oxide-superoxide donor on lipopolysaccharide-mediated accumulation of IL-8 from cultured human neutrophils. Interleukin-8 was measured in culture supernatant after 20 h using enzyme immunoassay. The combined nitric oxide-superoxide donor, 3-morpholinosydnonimine (SIN-1), dose-dependently decreased lipopolysaccharide-mediated IL-8 accumulation (P < 0.01). SIN-1 also decreased IL-8 accumulation from unstimulated neutrophils (P < 0.001). In contrast, the pure nitric oxide donor, 1,2,3,4-oxatriazolium 5 amino chloride (GEA-3162), increased stimulated IL-8 accumulation (P < 0.01) and also increased IL-8 accumulation in unstimulated cells (P < 0.002). Nitric oxide and superoxide have profound effects on IL-8. These results have important implications for the treatment of patients with acute lung injury with inhaled nitric oxide. PMID- 9215024 TI - Influence of 0.2 minimum alveolar concentration of enflurane on the ventilatory response to sustained hypoxia in humans. AB - To determine the influence of 0.2 minimum alveolar concentration (MAC) of enflurane on the time course of ventilation during sustained hypoxia, we studied 10 healthy adult volunteers with and without enflurane. The following design was used: end-tidal Po2 was maintained at 13.3 kPa for 8 min, at 6.7 kPa for 20 min and at 13.3 kPa for 8 min. End-tidal Pco2 was held constant throughout at 0.67 kPa above the subject's natural value. Control experiments were conducted with no hypoxia imposed. During the experiment subjects breathed via a mouthpiece from an automated gas mixing system which controlled end-tidal values. Enflurane reduced baseline (euoxic) ventilation from 20.9 (SEM 2.0) litre min-1 to 10.1 (1.0) litre min-1 (ANOVA, P < 0.001). Enflurane reduced the acute ventilatory response to hypoxia (AHVR) from 20.1 (3.3) litre min-1 to 5.0 (1.3) litre min-1 (ANOVA, P < 0.01), and the ventilatory off-response at cessation of hypoxia from 11.7 (2.4) litre min-1 to 1.8 (0.5) litre min-1 (ANOVA, P < 0.02). There was no significant difference in hypoxic ventilatory decline (HVD) without and with enflurane (8.9 (2.4) litre min-1 vs 5.5 (1.1) litre min-1; ANOVA, ns). These results confirm that 0.2 MAC of enflurane suppressed the acute ventilatory response to hypoxia, but had no significant effect on the subsequent ventilatory decline during sustained hypoxia. PMID- 9215026 TI - Effects of volatile anaesthetics on human neutrophil oxidative response to the bacterial peptide FMLP. PMID- 9215027 TI - Heterogeneous effects of protamine on human mast cells and basophils. AB - To investigate the mechanisms of anaphylactoid reactions to protamine, we have examined the in vitro effects of increasing concentrations of protamine (10(-6)-3 x 10(-4) mol litre-1) on the release of preformed (histamine and tryptase) and de novo synthesized (peptide leukotriene C4 (LTC4) or prostaglandin D2 (PGD2)) mediators from human basophils and mast cells isolated from lung parenchyma, heart, skin and synovial tissues. Protamine 10(-6)-3 x 10(-4) mol litre-1 induced release of histamine, but not de novo synthesis of LTC4 from basophils. At concentrations from 10(-5) to 3 x 10(-4) mol litre-1 it induced histamine release from human heart (mean 6.5 (SEM 1.5)%), skin (17.7 (4.1)%) and to a lesser extent from synovial mast cells, but not from lung mast cells. Protamine also caused the release of tryptase from heart mast cells (12.8 (3.2) micrograms/10(7) cells), but did not induce de novo synthesis of LTC4 and PGD2 from lung and skin mast cells. In these experiments cross-linking of IgE by anti-IgE caused release of LTC4 or PGD2 from human basophils or mast cells. These results demonstrate that protamine acted as an incomplete secretagogue, causing the release of preformed mediators from human basophils and mast cells. PMID- 9215028 TI - Glutamate: a role in normal brain function, anaesthesia, analgesia and CNS injury. PMID- 9215029 TI - Continuous extradural infusion of ropivacaine 2 mg ml-1 for pain relief during labour. AB - We have assessed the dose-response relationship of a solution of ropivacaine 2 mg ml-1, given as a continuous extradural infusion to women in labour. A total of 133 parturients were allocated randomly to one of four groups to receive a fixed rate ropivacaine infusion of 4, 6, 8 or 10 ml h-1 with additional bolus doses as necessary. Contraction pain, quality of analgesia, sensory block, motor block and neonatal Apgar scores were assessed. There were no significant differences between groups in terms of analgesia or motor block, although significantly more bolus doses were required by the group receiving 4 ml h-1 (P < 0.05 compared with the other groups), and a significantly higher total dose of ropivacaine was administered to the 10-ml h-1 group compared with the 6-ml h-1 group (P = 0.044). There were no significant differences between groups in terms of obstetric or neonatal outcome. We conclude that ropivacaine 2 mg ml-1 was effective and well tolerated when given as a continuous extradural infusion at 6-8 ml h-1 and may be used as the sole analgesic during labour. PMID- 9215031 TI - "Best" PEEP during one-lung ventilation. AB - Eight patients were studied under general anaesthesia for elective pulmonary lobectomy to see if intrinsic positive end-expired pressure (PEEPi) would appear or increase in the dependent lung during one-lung ventilation (OLV) or if application of external PEEP equal to individually measured PEEPi would produce better arterial oxygenation, haemodynamic state and oxygen delivery than either zero PEEP (ZEEP) or an external PEEP 5 cm H2O greater than PEEPi. Patients were non-obese, without obstructive airways disease, aged 53-76 yr and ASA < III. They received standardized anaesthesia with fentanyl, 50% nitrous oxide in oxygen and isoflurane; monitoring included radial and fibreoptic pulmonary arterial catheters and intermittent positive pressure ventilation with a tidal volume of 8 ml kg-1, 16 bpm, and an I:E ratio of 1:2. PEEPi was measured during two-lung ventilation (TLV) and OLV, using rapid airway occlusion at end-expiration. There was no PEEPi during TLV, but 2-6 mm Hg of PEEPi appeared during OLV. Applying external PEEP equal to individually measured PEEPi reduced venous admixture and increased PaO2 without a decrease in cardiac index (thus increasing oxygen delivery) compared with ZEEP, but the improvement in pulmonary gas exchange was lost and an additional penalty of reduced cardiac output was imposed when external PEEP was increased to 5 mm Hg above PEEPi. PMID- 9215030 TI - Processed electroencephalogram during combined extradural and general anaesthesia. AB - The processed electroencephalogram (pEEG) was monitored in eight patients undergoing gynaecological laparotomy under combined extradural and nitrous oxide isoflurane anaesthesia. Pre-incisional mean spectral edge frequency 95 percentile (SEF95) and median frequency (MF) were 11.67 (SD 1.63) Hz and 3.74 (0.24) Hz, respectively. After skin incision, both SEF95 and MF decreased to 6.61 (2.04) Hz and 2.72 (0.32) Hz, respectively (P < 0.001). An increase in mean arterial pressure after incision suggested inadequate depth of anaesthesia. After introduction of extradural analgesia, these variables returned to pre-incisional values (SEF95 11.65 (1.73); MF 4.02 (0.41)). Reduction of end-tidal isoflurane from 1.0% to 0.5% after extradural analgesia did not cause significant pEEG changes. pEEG may assist anaesthetists to recognize adequacy of combined general extradural anaesthesia. PMID- 9215032 TI - Antioxidant potential of i.v. fluids. AB - In acute clinical settings where there may be a role for antioxidant therapy, patients receive large amounts of i.v. fluids which may have antioxidant activity. To investigate such effects the antioxidant capacity of nine i.v. fluids was measured (Gelofusine, Haemaccel, 20% mannitol, 4.5% human albumin solution, fresh frozen plasma, aprotinin, N-acetylcysteine and two hydroxyethyl starch solutions). Results are expressed as mean mmol litre-1 Trolox equivalents. Mannitol 20% and the hydroxyethyl starch solutions had no antioxidant activity. Protein-containing solutions (gelatins, albumin and aprotinin) had antioxidant activity 50-66% that of plasma: Gelofusine 0.85 mmol litre-1; Haemaccel 0.78 mmol litre-1; 4.5% albumin 0.95 mmol litre-1; aprotinin 0.80 mmol litre-1; fresh frozen plasma 1.45 mmol litre-1. However, none was nearly as potent an antioxidant as the clinical preparation of N-acetylcysteine, with an antioxidant activity of 502 mmol litre-1. Studies of antioxidant therapy may need to take account of the antioxidant effect of i.v. fluids. PMID- 9215033 TI - Perioperative use of the oesophageal Doppler probe (ODM II) on a patient scheduled for transmyocardial revascularization. AB - We describe the use of the continuous wave oesophageal Doppler monitor (ODM II) in the perioperative management of a patient with chronic obstructive coronary artery disease undergoing transmyocardial revascularization (TMR). The use of ODM II allowed both quantitative and qualitative assessment of cardiac function relatively noninvasively. It detected the successful transmyocardial penetration of a laser beam during operation by visual and auditory phenomena in addition to reflecting improvement in cardiac performance after operation. PMID- 9215034 TI - Toward the paperless hospital? AB - This report describes an ethnographic study of document use by anaesthetists. In doing so, it focuses on the role of the preoperative risk assessment form as used in anaesthetic practice at a cardiothoracic hospital, and considers what would be the advantages and disadvantages of shifting the paper into the electronic form. Evidence from this case study is used to comment on how the practical use of documents by medical professionals can be fundamentally at odds with how the organization at large would like to use them. We argue that hospital trusts need to take into account this practical perspective in order to build effective, on line document systems. PMID- 9215035 TI - Guidelines for autologous transfusion. II. Perioperative haemodilution and cell salvage. British Committee for Standards in Haematology Blood Transfusion Task Force. Autologous Transfusion Working Party. PMID- 9215036 TI - Use of rocuronium in a pregnant patient receiving magnesium medication. PMID- 9215038 TI - Safety issues and volatile agent analysers. PMID- 9215037 TI - Hepatocellular integrity after inhalation anaesthesia. PMID- 9215039 TI - Ether in an isoflurane vaporizer and the use of vapour analysers in safe anaesthesia. PMID- 9215040 TI - Non-invasive measurement of cerebral blood flow. PMID- 9215041 TI - Non-invasive measurement of cerebral blood flow. PMID- 9215042 TI - Tolerance of laparoscopy for resection of phaeochromocytoma. PMID- 9215043 TI - Airway problems after carotid endarterectomy. PMID- 9215044 TI - In vivo and in vitro dose-response curves. PMID- 9215045 TI - Ambulatory extradural analgesia in labour and mode of delivery. PMID- 9215046 TI - The "Dumfries claim". PMID- 9215047 TI - Allergies and anaesthesia. PMID- 9215048 TI - The lens is more sensitive to radiation than we had believed. PMID- 9215049 TI - Radiotherapy for ocular angiomas. PMID- 9215050 TI - Osteoporosis: a survey of consultant ophthalmologists. PMID- 9215051 TI - A clinical study of radiation cataract formation in adult life following gamma irradiation of the lens in early childhood. AB - AIMS: To analyse long term effects on the lens of radium irradiation during infancy. METHODS: An infant cohort (n = 20, median age 6 months) treated for skin haemangioma with one or two radium-226 needles located at or within the orbital rim was examined 30 to 45 years after gamma radiation. Detailed information about the treatment procedure was available for all cases. Subcapsular opacities were graded semiquantitatively according to a scale based on extent and density of the opacities. RESULTS: A high prevalence of light to moderate posterior, subcapsular, and cortical cataract formation was found in the lenses on the treated side irradiated with a mean dose ranging from approximately 1 to 8 Gy. The cataract formation increased as a function of dose. The presence of subcapsular punctate opacities and vacuoles in the lenses on the untreated side receiving irradiation of an estimated dose varying around 0.1 Gy indicates a higher sensitivity than expected. CONCLUSION: The growing lens during infancy is sensitive to radium irradiation at doses lower than those previously stated. The eye lens seems suitable for studies of effects of low dose radiation since damaged cells are retained in the lens for a lifetime. PMID- 9215052 TI - Long-term results after low dose ocular irradiation for choroidal haemangiomas. AB - AIM/BACKGROUND: The most common choice of treatment for choroidal haemangiomas (CH) in the past has been the employment of scatter photocoagulation of the surface. This management often requires repetitive treatment or additional invasive management due to massive exudative detachment of the retina. The aim of this retrospective study was to investigate the outcome of the alternative application of low dose external beam irradiation with high energetic photons on these tumours. METHODS: A total absorbed dose of 20 Gy was applied to a total of 51 symptomatic eyes: 36 with a circumscribed CH of the posterior pole and 15 with diffuse CH as part of the Sturge-Weber syndrome. The indication for treatment was an exudative retinal detachment including or threatening the fovea. The mean follow up times in each group were 4.5 and 5.3 years, respectively. Out of a group of 33 patients from whom reliable data could be derived, 17 had symptoms lasting longer than 6 months. RESULTS: In 23 cases (63.8%) with circumscribed CH complete resolution of the subretinal fluid was achieved; the remaining 13 cases (36.2%) showed residual serous detachment distant to the fovea. The visual acuity improved by two or more lines in 14 cases (38.9%), remained stable in 14 cases (38.9%), and decreased in eight cases (22.2%). The functional success was dependent on the lag duration between onset of first subjective symptoms and treatment. The morphological results with diffuse CH were similar to those of the group of circumscribed CH. The visual acuity (VA) at last examination was improved in seven cases (46.6%); in the remaining eight cases, VA was unchanged or had deteriorated. The poor functional outcome in the latter was mainly attributable to secondary glaucoma. CONCLUSION: External beam irradiation is a useful and a low invasive therapeutic option for CH. A successful functional outcome is dependent on the time delay between first onset of symptoms and the beginning of therapy, the formation of subretinal fibrosis, and also on secondary glaucoma in the case of Sturge-Weber syndrome. PMID- 9215053 TI - Retinal vessel dilatation and elongation precedes diabetic macular oedema. AB - AIMS/BACKGROUND: Retinal vessel dilatation is a well known phenomenon in diabetes. In this study, the theory of whether excessive changes in diameter and length of retinal vessels occur in the development of diabetic macular oedema was tested, supporting a hypothesis that the development of diabetic macular oedema may be linked to hydrostatic pressure changes described in Starling's law. METHODS: From fundus photographs of diabetic patients attending a regular eye screening programme, the diameter and segment length of retinal vessels were measured in three retinopathy groups (12 patients each) with diabetic macular oedema (DMO), background retinopathy and no retinopathy, over a period of approximately 4 years, ending at the time of diagnosis of diabetic macular oedema in the DMO group. RESULTS: A statistically significant dilatation and elongation of retinal arterioles, venules, and their macular branches was found before the diagnosis of macular oedema in the DMO group. No significant changes were found in the other two groups. CONCLUSION: It is suggested that Starling's law applies to the formation of oedema in the retina as in other tissues. PMID- 9215054 TI - Altered retrobulbar vascular reactivity in early diabetic retinopathy. AB - AIM/BACKGROUND: In diabetic eye disease the factors leading to compromised circulation and the resulting loss of visual function are poorly understood. Although retinal circulation has been widely investigated, it accounts for only a fraction of total eye blood flow. Blood flow was investigated in the larger vessels feeding the eye in patients with early diabetic retinopathy. METHODS: Eleven patients with early diabetes with minimal or no retinopathy and 11 healthy controls were evaluated for retrobulbar blood flow velocity using colour Doppler imaging for the ophthalmic and central retinal arteries. Patients and subjects were tested while breathing room air and again under conditions of isocapnic hyperoxia. RESULTS: Hyperoxia induced a significant change in the central retinal artery end diastolic velocity (EDV) (p = 0.008) and resistance index (RI) (p = 0.032) in normal subjects, but not in diabetic patients. Consequently, during hyperoxia, the diabetic patients were significantly higher for EDV (p = 0.006) and significantly lower for RI (p = 0.002) compared with normal controls. Hyperoxia caused no significant change in either group in the ophthalmic artery; nevertheless, under isocapnic hyperoxia conditions the diabetic patients had lower peak systolic velocity (p = 0.05) and lower RI (p = 0.05) than normal subjects. CONCLUSIONS: Imposition of isocapnic hyperoxia produces significant differences in the ophthalmic and central retinal artery blood flow velocities in diabetic patients with early disease when compared with normal subjects. These results demonstrate that diabetic patients with minimal or no retinopathy suffer from irregular ocular vascular function in the major vessels feeding the eye. PMID- 9215055 TI - Retinopathy of prematurity: systemic complications associated with different anaesthetic techniques at treatment. AB - BACKGROUND: Treatment of retinopathy of prematurity (ROP) in the UK is subject to considerable regional variation in terms of anaesthetic support. Change in practice at St Mary's neonatal medical unit from topical to general anaesthesia and, subsequently, to sedation/analgesia allowed comparison of the impact of these three modalities on infants' early postoperative course in a consecutive, non-randomised, observational study. METHODS: The study population consisted of 30 babies undergoing treatment of threshold ROP. Twelve were treated using topical anaesthesia alone (group A), six using general anaesthesia (group B), and 12 using sedation/analgesia combined with elective intubation and artificial ventilation (group C). Daily measurements of infant health were recorded starting 4 days preoperatively and continuing for 7 days postoperatively to facilitate the formulation of a cardiorespiratory stability index as follows: (0) improvement from baseline, (1) no change from baseline, (2) mild instability, (3) marked instability, and (4) life threatening event. RESULTS: Within the first 48 hours postoperatively in group A 5/12 showed mild instability and 4/12 showed marked instability (including three babies suffering life threatening events requiring emergency resuscitation). In group B within the first 48 hours postoperatively 1/6 showed mild and 1/6 showed marked instability, and in group C 5/12 babies showed mild instability alone. There was a significant difference for cardiorespiratory stability scores between the three groups overall for the 7 days postoperatively (repeated measures ANOVA, p = 0.018). CONCLUSIONS: Premature infants undergoing cryotherapy for ROP who were treated using topical anaesthesia alone had more severe and protracted cardiorespiratory complications. PMID- 9215056 TI - Topical anaesthesia with oxybuprocaine versus sub-Tenon's infiltration with 2% lignocaine for small incision cataract surgery. AB - AIMS: To determine whether topical anaesthesia in small incision self-sealing phacoemulsification cataract surgery provides comparable anaesthesia to sub Tenon's infiltration. METHODS: Thirty five patients undergoing small incision self-sealing phacoemulsification cataract surgery were allocated randomly to receive topical anaesthesia with 0.4% oxybuprocaine or sub-Tenon's infiltration with 2% lignocaine. Pain experienced during the operation was assessed by asking the patient to score on a visual analogue graphic pain score chart. RESULTS: The median pain score for the topical group (3) was significantly higher than that of the sub-Tenon's group (0) (p = 0.004). CONCLUSION: Sub-Tenon's infiltration is superior to topical anaesthesia in ensuring patient comfort during small incision scleral tunnel self-sealing phacoemulsification cataract surgery. PMID- 9215057 TI - Retinal dystrophy in long chain 3-hydroxy-acyl-coA dehydrogenase deficiency. AB - BACKGROUND: Long chain 3-hydroxyacyl-CoA-dehydrogenase (LCHAD) is one of the enzymes involved in the breakdown of fatty acids. A deficiency of this enzyme is associated with life threatening episodes of hypoketotic hypoglycaemia during prolonged fasting. Neuropathy and retinopigmentary changes were mentioned in only a few cases. METHODS: The case histories of two girls, aged 8 and 15 years, with LCHAD deficiency are reported. RESULTS: Both children with LCHAD deficiency exhibited extensive macular pigmentary depositions and a 'salt and pepper' scattering of pigment in their retinas. The patients have decreasing visual acuity. CONCLUSION: The early recognition of LCHAD deficiency can increase the life expectancy in these patients through avoiding catabolism and through appropriate diets. Patients tend to be free of symptoms between attacks, however. Testing for the disorder, therefore, should be included in the diagnostic process for children with retinal dystrophy, in particular when other clinical symptoms are known to have occurred. PMID- 9215058 TI - Ocular manifestations of familial amyloidotic polyneuropathy type I: long-term follow up. AB - AIMS: To obtain precise information on ocular manifestations of familial amyloidotic polyneuropathy (FAP) type I, the incidence of five main ocular manifestations--abnormal conjunctival vessels (ACV), keratoconjunctivitis sicca (KCS), pupillary abnormality, vitreous opacity, and glaucoma, were compared through long term follow up. METHODS: Ocular examinations were performed in 37 FAP type I patients (Met30) from once to 12 times over a period of 1 to 12 years and 7 months. RESULTS: The following incidences were observed on initial examination of each patient with FAP: ACV in 75.5%, pupillary abnormalities in 43.2%, KCS in 40.5%, glaucoma in 5.4%, and vitreous opacity in 5.4%. All ocular manifestations increased with the progression of FAP, and the incidence of ACV reached 100% during follow up: this may be helpful in the diagnosis of FAP. CONCLUSION: Since no precise statistical ocular study on FAP with long term follow up has been performed, this report may provide important information to help elucidate the mechanism of the amyloid distributing process in the amyloid targeted organs of FAP and to provide the natural course of ocular manifestations of FAP. PMID- 9215059 TI - Comparison of the efficacy of diclofenac and betamethasone following strabismus surgery. AB - AIMS: To compare the relative antiinflammatory potency and safety of topical diclofenac-gentamicin with betamethasone-neomycin following strabismus surgery. METHODS: A single centre, single observer, prospective, randomised, and double masked clinical trial of 25 children undergoing bilateral symmetrical horizontal strabismus surgery was carried out. One eye received diclofenac-gentamicin and the contralateral eye received betamethasone-neomycin; both treatments were instilled four times a day for 4 weeks postoperatively. Ocular inflammation was assessed at 1 and 4 weeks postoperatively, objectively by comparison with a photographic chart and subjectively by questionnaire. RESULTS: There was no statistically significant difference in the rate of resolution of the inflammatory response between each group at both visits. CONCLUSION: Diclofenac appears to be as effective as betamethasone in controlling postoperative inflammation following strabismus surgery and may offer a safer alternative to the use of topical steroids. PMID- 9215060 TI - Technique of goniocurettage: a potential treatment for advanced chronic open angle glaucoma. AB - AIM: To introduce a new concept of anterior chamber angle microsurgery, designed to scrape pathologically altered trabecular meshwork from the scleral sulcus as a potential treatment in primary open angle glaucoma. METHODS: Gonioscopically controlled ab interno abrasion of the trabecular meshwork was performed on six human eye banking eyes for morphological analysis. Thereafter, four eyes suffering from terminal glaucomatous optic nerve atrophy as a result of medically uncontrolled intraocular pressure were also treated by 'goniocurettage'. The newly designed instrument resembles a modified cyclodialysis spatula with a bowl shaped tip, 300 microns in diameter, and with its edges sharpened. The treatment zone comprised 4-5 clock hours of the chamber angle circumference. RESULTS: Microscopic examination of the treatment zone revealed that in addition to a complete disruption of the trabecular meshwork and internal wall of Schlemm's canal goniocurettage also caused damage to intracanalicular septa. A splitting along the posterior wall of Schlemm's canal was also noted in one specimen. The clinical data of goniocurettage also showed some promising results. Mean pretreatment IOP averaged 40.7 (SD 8.8) mm Hg (range 32-51 mm Hg) and was significantly (p < 0.04) reduced to 18.0 (4.2) mm Hg (12-22 mm Hg) after 6 months, representing an absolute decrease in IOP of 22.7 mm Hg and a mean decrease in IOP of 56%. Clinically significant hyphaema occurred in one eye, caused by introgenic trauma to a prominent chamber angle vessel. In three eyes a minor reflux of blood occurred at the treatment site. However, no hypotony, choroidal effusion, flattened anterior chamber, or cyclodialysis were observed in these patients. CONCLUSION: Morphological analysis of treated postmortem eyes confirmed that goniocurettage completely removed the trabecular meshwork and opened Schlemm's canal, ensuring direct access into the anterior chamber. In a small number of patients over a limited period of time this new surgical procedure resulted in a clinically significant pressure reduction. However, longer term follow up and a greater number of patients are warranted before this experimental procedure is applicable to eyes that would do well with conventional surgery. PMID- 9215061 TI - Detection of human papillomavirus infection in squamous tumours of the conjunctiva and lacrimal sac by immunohistochemistry, in situ hybridisation, and polymerase chain reaction. AB - BACKGROUND: Squamous tumours of the ocular surface, including the lacrimal pathway, range from benign lesions to invasive carcinomas. Some of these tumours are associated with human papillomavirus (HPV) infection, with the types of HPV differing among papillomas and dysplastic or malignant lesions. METHODS: The relation between squamous tumours of the conjunctiva and lacrimal sac and HPV infection was investigated in 17 individuals with such tumours. Nine of the 17 tumours were benign, four were dysplastic lesions, and four were carcinomas. RESULTS: Eight specimens showed positive immunohistochemical staining with antibodies to HPV; four of these eight were papillomas, three were dysplastic lesions, and one was a carcinoma. Koilocytosis was detected in seven of these eight tumours. Five of the eight specimens positive for immunohistochemical staining were also positive for HPV DNA by in situ hybridisation, and all eight were positive for HPV DNA by the polymerase chain reaction (PCR) method. CONCLUSION: Approximately 50% of squamous tumours of the ocular surface and lacrimal sac were associated with HPV infection. This is the first report, to our knowledge, of the detection of HPV in the field of ophthalmology by a combination of immunohistochemistry, in situ hybridisation, and PCR. PMID- 9215063 TI - First report of congenital or infantile cataract in deranged proteoglycan metabolism with released xylose. AB - AIM: To investigate the chemical pathology in the blood and lens, in cases of congenital or infantile cataract in children excreting predominantly non-reducing carbohydrates in urine. METHODS: Urine samples from children with congenital or infantile cataract, and age and sex-matched controls, were analysed for (i) inherited errors of metabolism, (ii) paper chromatography of sugars, (iii) spectrophotometric assay of glycosaminoglycans (GAG), (iv) cetyl trimethyl ammonium bromide test, (v) electrophoresis using Alcian blue, (vi) ion exchange chromatography with IR 120 resin, and (vii) HPLC for xylose. Blood and lens material were also tested for GAG fragments and xylose. beta Glucuronidase was assayed in lymphocytes and urine. RESULTS: Of 220 children of both sexes below 12 years of age, with congenital or infantile cataract treated in Sankara Nethralaya, Madras, India, during a period of 2 years, 145 excreted fragments of GAG (heparan and chondroitin sulphates) in their urine. There was no such excretion among the control group of 50 children. The same was found accumulated in the blood and lenses of affected children. In addition, xylose was present in small amounts in the urine and blood and xylitol was present in the lens. There was a significant elevation in the activity of beta glucuronidase in lymphocytes and urine, when compared with normals. All the above findings suggest deranged proteoglycan metabolism. As the urine contained mostly GAG fragments and very little xylose, Benedict's reagent was not reduced. This ruled out galactosaemia. CONCLUSION: An increase of beta glucuronidase activity might have caused extensive fragmentation of GAG with resultant accumulation in the blood and lens and excretion in urine. Small amounts of xylose may have come from xylose links between GAG and core protein of proteoglycans. Owing to their polyanionic nature, GAG fragments in the lens might abstract sodium, and with it water, thereby increasing the hydration of the lens. Excessive hydration and the osmotic effect of xylitol from xylose might cause cataract. While corneal clouding has been reported in inborn acid mucopolysaccharidosis, congenital or infantile cataract with deranged metabolism of proteoglycans (acid mucopolysaccharide-xylose-protein complex) is reported in children for the first time. PMID- 9215062 TI - Antioxidant enzymes in the human iris: an immunogold study. AB - AIMS: To determine the nature and extent of the antioxidant enzyme system in the human iris. METHODS: The fine structural distribution of five antioxidant enzymes (acidic, neutral, and basic glutathione S-transferase (GST), Cu/Zn superoxide dismutase (Cu/Zn SOD), and glutathione peroxidase) was determined by immunogold labelling of ultrathin sections of tissue from six eyes appropriately fixed for immunocytochemistry and embedded in LR white resin. RESULTS: Both Cu/Zn SOD and acidic GST were localised to all constituent cells of the iris. Glutathione peroxidase and basic GST were localised to erythrocytes alone, and labelling for neutral GST was absent. CONCLUSIONS: Dense labelling for acidic GST could be linked with the previously documented presence of large quantities of polyunsaturated fatty acids in the iris and/or the presence of xenobiotic substances in the aqueous humour. PMID- 9215065 TI - Long-term effect of repeated hyperbaric oxygen therapy on visual acuity in inflammatory cystoid macular oedema. PMID- 9215064 TI - Can visual function be restored in patients with homonymous hemianopia? PMID- 9215066 TI - Sarcoidosis presenting as a cutaneous eyelid mass. PMID- 9215067 TI - Orbital metastasis from carcinoma of cervix. PMID- 9215068 TI - 'Pogs and slammers': ocular injury caused by a new game. PMID- 9215069 TI - Canalicular stenosis in the course of primary herpes simplex infection. PMID- 9215070 TI - Ocular and cerebral trauma in non-accidental injury in infancy. PMID- 9215071 TI - Analysis of high-energy phosphometabolites in delayed experimental skin flaps using 31P nuclear magnetic resonance spectroscopy. AB - Using 31phosphorus magnetic resonance spectroscopy (31P-MRS) and surface coils, we noninvasively assessed the intracellular changes in delayed skin flaps of the high-energy phosphometabolites, ATP and phosphocreatine, which are basic energy sources of living cells. In 5 rats, a 3.5 x 7.5 cm bipedicled skin flap was elevated from the dorsum and its caudal base divided after a delay period of 2 weeks. MRS spectra were collected at four times: immediately, 1 week and 2 weeks after elevation of the bipedicled flap, and 18 hours after division of its caudal base. From the spectra, we calculated the intracellular pH and the following ratios: Pi/PCr, PCr/(PCr + Pi), ATP/(PCr + Pi) (PCr, phosphocreatine; Pi, inorganic phosphate; ATP, adenosine triphosphate). As an undelayed control group, cranially based skin flaps of the same size were elevated in another 5 rats, and MRS spectra were obtained 18 hours later. The length of the surviving area was longer in the delayed flaps than in the undelayed flaps. Intracellular pH and ATP/(PCr + Pi) showed no significant alteration in the delayed skin flaps, not only during the delay period but also after conversion of the flaps into cranially based flaps by division of their caudal base. In contrast, PCr/(PCr + Pi) decreased with each surgical procedure (bipedicled flap elevation and base division). Compared with the necrotic distal portion of the undelayed flaps, the surviving distal portion of the delayed flaps had higher levels of intracellular pH and ATP/(PCr + Pi) and lower levels of PCr/(PCr + Pi). Intracellular pH and ATP/(PCr + Pi) showed a strong correlation with the viability of the delayed and undelayed skin flaps, and they can be prognostic indices for predicting the fate of skin flaps. The reason the surviving distal portions of the delayed flaps maintained their level of ATP despite the low intracellular level of PCr may be that the accumulation of mitochondrial creatine kinase enhances the so-called 'energy transport' function of the creatine kinase/phosphocreatine system. PMID- 9215072 TI - The free 'V': a bipennate free flap for double eyelid resurfacing based on the second dorsal metacarpal artery. AB - We present a patient who had a basal cell carcinoma of the left upper eyelid and ectropion of the left lower eyelid. The patient underwent resection of the tumour and release of the ectropion resulting in a full thickness defect of the skin of his left upper and lower eyelids. The eyelids were reconstructed with a second dorsal metacarpal artery free flap from the left hand. For safety, a dorsal vein of the flap was arterialised and one of the valves of the vein had a valvotomy. The flap survived completely. PMID- 9215073 TI - The distally based islanded dorsal foot flap. AB - A modification of the distally based first web space flap on the dorsum of the foot is described. This flap, originally described by Earley and Milner, has been completely islanded on its blood supply and used to resurface defects following release of post-burn hyperextension contractures of the toes. Our experience with 14 consecutive flaps is presented. Partial necrosis occurred in three flaps, one leading to slight residual contracture. We have found this flap reliable and easy to raise, and the donor site morbidity is minimal. PMID- 9215074 TI - Free radial forearm flap to the foot after a shotgun injury: the day I became a patient. AB - This is a personal account of a General Practitioner who suffered a shotgun injury to his right foot. Reconstruction involved the use of a free radial forearm flap to fill a defect distal to the heel. The case is illustrated and the feelings of the patient described. PMID- 9215075 TI - Video microsurgery: a substitute for the operating microscopy? PMID- 9215076 TI - The breast fish. PMID- 9215077 TI - The distally based superficial sural flap for reconstruction of the lower leg and foot. PMID- 9215078 TI - Regeneration of sudomotor and sensory nerve fibres after digital replantation and microneurovascular toe-to-hand transfer. AB - The end-stage sudomotor and sensory recovery in patients with replanted fingers and patients after microneurovascular toe-to-hand transfer was studied using quantitative electrophysiological investigations (recovery of sensory nerve action potentials and the sympathetic skin response), the ninhydrin test and clinical testing of sensory regeneration (light touch, pain, static and dynamic two-point discrimination). 13 adult patients with 22 replanted digits (11 males, 2 females) aged 21-58 years (mean 42.2 years) and 12 adults and adolescents (8 males, 4 females) aged 13-45 years, (mean 26.8 years) following 14 microneurovascular great and/or second toe-to-hand transfers were studied. The replanted fingers were examined 2-7 years after injury and replantation. The toe to-hand transfers were examined 2-12 years after injury and transfer. The results show better end-stage recovery of sudomotor and sensory function following finger replantation when compared to microneurovascular toe-to-hand transfer. PMID- 9215079 TI - Comparative biomechanical analysis of a new circumferential flexor tendon repair and a modified Kessler repair. AB - We present the technical details and the results of a biomechanical analysis of a new type of circumferential flexor tendon repair, designed with the more stringent requirements of zone II injuries in mind. Apart from good initial strength we aimed for a design with little bulk at the repair site and good control of the tendon edges. The new repair is achieved using a single, continuous, inverting and locking suture of the periphery of the tendon. The repair was compared with a Kessler core suture of 4/0 polydioxanone, with Tajima and Strickland modifications, to which has been added a simple running circumferential suture (6/0 polypropylene), the repair currently used in our unit. Fresh human cadaver flexor tendons were divided and repaired by one of the two techniques (n = 12 for each technique), using 5/0 polypropylene for the new circumferential suture. A third group of tendons (n = 8) were divided and repaired with a 5/0 multifilament steel circumferential suture. The repaired tendons were tested at longitudinal stress to failure. The first two groups of tendons were tested at two crosshead speeds. Overall, crosshead speed had no effect on ultimate tensile strength (P = 0.5). The 5/0 polypropylene circumferential repair (median 32.29 N) was significantly stronger than the Kessler repair (median 24.03 N) (P = 0.046). The circumferential repair was significantly stronger with steel (median 56.04 N) than with polypropylene (median 32.29 N) (P = 0.007). The size of the repair site, resistance to gap formation and the patterns of failure were analysed on video recordings. PMID- 9215080 TI - Measuring patient-based outcomes in a plastic surgery service: breast reduction surgical patients. AB - On admission for breast reduction surgery, 110 patients completed a preoperative assessment pack containing: 1) Personal and demographic questions; 2) Condition specific questions including physical symptoms and areas of life affected by their condition; 3) The SF-36 Health Survey Questionnaire; and 4) The Rosenberg Self-esteem Scale. At 3 months and again at 6 months after surgery, these same patients were sent postal follow-up packs containing the SF-36 and Rosenberg questionnaires and postoperative condition-specific questions requesting information on complications, relief of physical symptoms, scarring, pain and opinion of the aesthetic result. The response rate was 82% (90 patients) at 3 months and 76% (84 patients) at 6 months. The results of the data collected indicate that breast reduction surgery confers very substantial benefit to patients in terms of greatly improved physical and psychological health and well being. PMID- 9215081 TI - The cutaneous innervation of the female breast and nipple-areola complex: implications for surgery. AB - Many surgical procedures performed in the thoracic region can easily damage cutaneous nerves important for the sensory innervation of the female breast. A better understanding of the distribution of these cutaneous nerves will help prevent impaired sensation after breast surgery. Therefore an anatomical study was performed on the cutaneous innervation of 12 breasts of 7 female cadavers. Special emphasis was placed on the nipple-areola complex. The origin, course and final destination of each cutaneous nerve was established and the contribution of each branch was determined by the area it innervated. Differences were evaluated using analysis of variance. The cutaneous innervation of the female breast is derived medially from the anterior cutaneous branches of the Ist-VIth intercostal nerves and laterally from the lateral cutaneous branches of the IInd-VIIth intercostal nerves. The nipple-areola complex is consistently supplied by the anterior and lateral cutaneous branches of the IVth intercostal nerve, with additional innervation by cutaneous branches of the IIIrd and Vth intercostal nerves. This study shows an equal importance of both the anterior and the lateral cutaneous branches of the intercostal nerves. During surgical procedures one should try to avoid damage to the anterior and lateral cutaneous branches of the IIIrd, IVth and Vth intercostal nerves, with special attention to the IVth intercostal nerve which is the consistent nerve to the nipple-areola complex. PMID- 9215082 TI - Vasoconstrictor infiltration in breast reduction surgery: is it harmful? AB - The efficacy of vasoconstrictor infiltration in reducing blood loss is well known. However, adrenaline infiltration is potentially harmful to tissues. The question of whether or not adrenaline infiltration is harmful in breast reduction surgery remains unanswered. We retrospectively reviewed the notes of 100 consecutive cases after bilateral breast reduction (n = 200 breasts) with preoperative infiltration of a vasoconstrictor solution (10 ml adrenaline 1:10,000, 20 ml lignocaine 1%, 70 ml saline 0.9%, resulting in an adrenaline concentration of 1:100,000), looking specifically at postoperative complications that could be secondary to adrenaline infiltration. Two breasts developed a 'haematoma'; both were of small volume. Six breasts developed a 'minimal wound' problem which involved the T-junction. 'Wound breakdown' was noted in five breasts and again involved the T-junction in most cases. 'Wound infection' occurred in eleven breasts. There was one case of partial 'nipple necrosis'. Complications occurred in 12.5% of breasts and 21% of patients. We feel that this complication rate is within acceptable limits. PMID- 9215083 TI - Giant lipoma of the breast. AB - Lipomas of the breast are usually small, benign neoplasms which can be treated by simple excision. Diagnosis of these masses, however, can be difficult because of the normal fatty composition of the breast. A number of radiological manoeuvres have been described for diagnosing deep lipomas in the breast, yet the clinical and radiographic identification of these masses remains challenging. We present a case of giant lipoma of the breast which was not appreciated on initial clinical and radiographic evaluation, and which was later found to comprise most of the mass of the breast. Following resection, the deformed breast was reconstructed using folded dermoglandular flaps from the areas expanded by the lipoma with excellent results. This case is an excellent illustration of the difficulties associated with diagnosis of these tumours and the reconstructive options available following excision. PMID- 9215084 TI - Recipient bed vascularity and the survival of ischaemic flaps. AB - The purpose of this study was to identify the length of the ischaemic period required to induce the 'no-reflow' phenomenon in a rat epigastric flap on an avascular recipient site. The vascularity of the recipient bed may affect flap survival in the early postischaemic stage after flap transfer. Initially, we designed epigastric flaps in 300-350 g Sprague-Dawley rats and separated the rats into four groups of 5 rats each (total 20 rats). In groups 1, 2, 3, the flaps were made ischaemic for 1 hour, 6 hours and 10 hours, respectively, by temporarily clipping the epigastric artery and vein. In group 4, the epigastric artery and vein were divided to create permanent ischaemia. In groups 1, 2 and 3, ischaemia was ended by removing the clips. After the ischaemic flaps were reperfused, their viability was studied by measuring the flap survival rate at postoperative day 7. Flap survival was studied by direct observations, laser Doppler flowmeter measurement of flap blood flow, histopathology, and carbon particle perfusion of the flap vasculature. Ischaemic flaps of groups 1 and 2 recovered almost completely after reperfusion due to the short period of ischaema. In a second series of experiments, in order to evaluate the contribution to flap survival of the recipient vascularised bed, another four groups of epigastric flaps (of 5 animals each, using the same time periods as above) were raised and a piece of Biobrane was interposed between the flap and the recipient bed before the flap wound was closed, to eliminate all nutrient supply from the recipient bed. THe results showed that the combined effect of the reperfused flap vasculature plus the metabolic contribution of the recipient bed significantly (P < 0.01) increased the extent of flap survival of the 6- and 10 hour ischaemic flaps as well as the divided pedicle flaps, which were never reperfused. An absolute 'no-reflow' rat model flap for further flap salvage studies was also developed. PMID- 9215085 TI - Beyond efficacy: new issues for HSV antiviral therapy. PMID- 9215086 TI - Diagnostic utility of bone marrow sampling in HIV positive patients. PMID- 9215088 TI - Is Europe ready for STD screening? PMID- 9215087 TI - Health education and promotion for STD prevention: lessons for the next millennium. AB - OBJECTIVE: To review the evolution of health promotion for STD prevention. MAIN OBSERVATIONS: Information and education programmes were provided at the beginning of the 20th century to warn the public about the dangers of venereal infection and to support the medical model of case identification and case management under the care of qualified physicians. The public health approach offered advice about chemical, chemotherapeutic, and barrier prophylaxis, but avoided the issue of social prophylaxis. With the failure of antimicrobial agents to eradicate syphilis in the 1960s, rapid increases of viral sexually transmitted diseases (STDs) and resistant strains of gonorrhoea in the 1970s, and the discovery of AIDS in the 1980s, alternatives to the traditional public health approach were sought and supported with a modest increase of resources. Three major innovations have been introduced to STD prevention as a result: social marketing, community involvement, and behaviour change programmes based on social and psychological concepts and theoretical models. CONCLUSIONS: Health promotion for STD prevention in the future will be characterised by careful assessments of the social and behavioural determinants of sexual risk taking, development and implementation of targeted interventions designed to reduce risk taking, and evaluation of social and behavioural interventions for improvements in STD prevention. PMID- 9215089 TI - Chlamydia screening: which sample for which technique? PMID- 9215090 TI - Is screening for Chlamydia trachomatis infection cost effective? PMID- 9215091 TI - Valaciclovir for the suppression of recurrent genital HSV infection: a placebo controlled study of once daily therapy. International Valaciclovir HSV Study Group. AB - OBJECTIVE: To determine the efficacy and safety of once daily valaciclovir for the suppression of recurrent genital herpes simplex virus (HSV) infection in immunocompetent patients. METHODS: 382 otherwise healthy patients with a history of frequently recurring genital HSV infection (eight recurrences per year) were randomly allocated to receive either oral valaciclovir (500 mg once daily) or placebo (3:1 ratio) for 16 weeks or until the first genital HSV recurrence, whichever occurred first. Patients were clinically assessed at regular intervals and also if they experienced a recurrence. Safety was evaluated through adverse experience reporting and monitoring of haematology and biochemistry variables. On completion of the double blind phase, patients were eligible for follow up to a maximum of 48 weeks' treatment with open label valaciclovir (500 mg once daily) for further safety monitoring. The results from the double blind phase of the study are reported here. RESULTS: A significant difference was detected between valaciclovir and placebo in the time to first recurrence of genital HSV infection. The hazard ratio [95% confidence interval] for valaciclovir v placebo was 0.155 [0.112, 0.214], p < 0.0001. Valaciclovir prevented or delayed 85% of the recurrences that would have occurred with placebo. After 16 weeks (day 112) with treatment, 69% of patients receiving valaciclovir were recurrence free compared with only 9.5% of patients assigned to placebo. The safety profiles of valaciclovir and placebo were comparable, with adverse experiences being infrequent and generally mild. CONCLUSION: This study has demonstrated that once daily valaciclovir (500 mg), is highly effective and well tolerated for the suppression of recurrent genital HSV infection. Once daily dosing with valaciclovir provides a more convenient dosing regimen than the more frequent aciclovir regimens. PMID- 9215093 TI - Diagnostic utility of bone marrow sampling in HIV positive patients. AB - OBJECTIVE: To evaluate the diagnostic utility of bone marrow (BM) sampling in HIV positive patients. DESIGN: Retrospective cohort analysis. SETTING: Specialist HIV/AIDS service in London. SUBJECTS: 215 consecutive HIV infected patients undergoing 246 BM samplings for investigation of pyrexia without localising signs, haematological abnormalities, or staging/investigation of lymphoma. MAIN OUTCOME MEASURE: Diagnostic yield from (and impact on management of) BM sampling. RESULTS: Of 122 BM samples taken to investigate pyrexia, 33 (27%) revealed the cause on microscopy: unexpected lymphoma in seven (6%), mycobacteriosis in 25 (20%), and toxoplasmosis in one (1%). Marrow infiltration was confirmed in 11 of 38 BM samples taken for staging/investigation of lymphoma/leukaemia. In afebrile patients, of 22 with pancytopenia, BM samples showed HIV associated changes in 17 and specific diagnoses in five (mycobacterial infection in three, haemophagocytic syndrome in one, and megaloblastic change due to vitamin B-12 deficiency in one); of 21 with isolated thrombocytopenia, 20 (95%) BM samples showed immune thrombocytopenic purpura to be the cause and the remaining patient had BM changes of aplasia; of 29 with isolated anaemia, 28 had BM changes of HIV associated dysplasia/erythroid dysplasia and one had unsuspected iron deficiency; all 10 with isolated leucopenia/neutropenia had BM changes ascribed to HIV infection exacerbated by concurrent sepsis or medication; of four BM samples taken for other reasons, one showed mycobacterial infection. CONCLUSIONS: BM sampling has diagnostic utility in HIV infected patients with pyrexia without localising signs, pancytopenia, and staging/investigation of lymphoma; this test has little value in the investigation of afebrile patients with isolated thrombocytopenia, anaemia, or leucopenia as HIV is usually the underlying cause. PMID- 9215094 TI - Prison doctors beware!. PMID- 9215092 TI - Valaciclovir versus aciclovir in patient initiated treatment of recurrent genital herpes: a randomised, double blind clinical trial. International Valaciclovir HSV Study Group. AB - OBJECTIVE: To compare the efficacy and safety of twice daily valaciclovir with five times daily aciclovir in the treatment of an episode of recurrent genital herpes simplex virus (HSV) infection in immunocompetent individuals. METHODS: 739 patients with a history of recurrent genital HSV infection received either oral valaciclovir (500 mg twice daily) or aciclovir (200 mg five times daily) for 5 days for treatment of their next recurrent episode in a controlled, randomised, double blind trial. Patients self initiated therapy at the first signs and/or symptoms of the HSV recurrence, then were assessed in clinic on five occasions over 7 days, and twice weekly thereafter until lesions had healed. Safety was evaluated through adverse experience reports and haematology and biochemistry monitoring. RESULTS: No significant differences were detected between valaciclovir and aciclovir for the primary endpoint, the duration of all signs and symptoms which included lesion healing and pain/discomfort. The hazard ratio [95% confidence interval] for valaciclovir v aciclovir was 0.93 [0.79, 1.08]. Lesion healing time was similar in each treatment group (hazard ratio valaciclovir v aciclovir 0.96 [0.80, 1.14]). The odds ratio of valaciclovir v aciclovir in preventing the development of vesicular/ulcerative lesions was 1.08 [0.82, 1.42]. Percentages of patients in whom all HSV cultures were negative were similar in the valaciclovir and aciclovir groups at 59% and 54% respectively; for patients having equal to or more than one positive culture result after treatment initiation, cessation of virus shedding was similarly rapid for the two treatments (hazard ratio 0.98 [0.75, 1.27]). The safety profiles of valaciclovir and aciclovir were comparable with adverse experiences being infrequent and generally mild. CONCLUSION: This study has demonstrated that valaciclovir 500 mg twice daily is equivalent in efficacy to aciclovir 200 mg five times daily as episodic treatment of recurrent genital HSV infection. Valaciclovir maintains the established efficacy and safety of aciclovir but offers a much more convenient twice daily dosing regimen. PMID- 9215095 TI - Epidemiology of genital Chlamydia trachomatis in England and Wales. AB - OBJECTIVE: To describe the recent epidemiology of genital Chlamydia trachomatis infection in England and Wales. DESIGN: Retrospective study of routinely available surveillance datasets and ad hoc prevalence studies. METHODS: Numbers of new cases of genital C trachomatis infection, obtained from the Department of Health and Welsh Office, were combined with the estimated mid-year resident population of England and Wales. Rates were analysed for trend over time using a log linear age period model in GLIM4. Ad hoc prevalence and case finding studies carried out over the past 20 years were critically assessed in terms of study design and testing methodologies. RESULTS: Attendance rates at genitourinary medicine (GUM) clinics were higher for women than men over the period 1989 to 1994 as were the number of laboratory reports. The highest rate of attendance (GUM clinic data) was for women aged 16 to 19 years. There was an overall significant linear decrease in the attendance rates over time for both men (p = 0.0172) and women (p = 0.0000) between 1989 and 1994. There was considerable variation in the prevalence of genital C trachomatis infection detected within different clinical settings, together with a substantial level of asymptomatic infection. CONCLUSIONS: Genital C trachomatis infection is broadly distributed throughout the sexually active population, with a substantial reservoir of asymptomatic infection among those generally perceived to be at low risk of a sexually transmitted infection. Young people, particularly women aged 16 to 19 years, are at highest risk of genital C trachomatis infection. This is of concern since younger women are more susceptible than older women to developing complications of chlamydial infection, such as pelvic inflammatory disease. The broad distribution of infection across all sexually active health service attenders and the high level of asymptomatic infection suggest that a new, screening based, approach to the control of genital C trachomatis infection is required. Recommendations are made as to the epidemiological research required to guide such work. PMID- 9215096 TI - Hepatitis B markers in heterosexual patients attending two genitourinary medicine clinics in the West Midlands. AB - OBJECTIVE: To determine the prevalence of hepatitis B virus (HBV) infection in heterosexual patients attending two genitourinary medicine (GUM) clinics in the West Midlands and to examine whether heterosexual activity is a risk factor for acquiring HBV infection with the view to extend HBV vaccination policies to cover this group. DESIGN: HBV markers were determined in the GUM study group and compared with that of the control groups. Responses to a questionnaire were used to examine sexual behaviour patterns that may be related to heterosexual acquisition of HBV infection. SETTING: The West Midlands, UK April 1992-January 1993. SUBJECTS: 788 male patients and 688 female patients attending GUM clinics were compared with 498 male blood donors and 563 females attending antenatal clinics for the seroprevalence of HBV markers. Potential risk factors related to heterosexual activity were assessed in 1436 patients in the study group. MAIN OUTCOME MEASURES: Prevalence of HBV markers in the GUM study group and the controls. The possible use of the risk factors examined as predictors for acquiring HBV infection. RESULTS: The seroprevalence of hepatitis B core antibody (anti-HBc) in GUM patients was 1.9% and 0.5% in the control group. In the study groups the prevalence of anti-HBc from Birmingham was 3.2% while that from Coventry was 0.8%. The low seroprevalence of HBV prevented a multiple logistic analysis. A limited regression analysis showed that being non-white (p < 0.001) and duration of sexual activity (p = 0.013) were risk factors for HBV infection. However, these two factors were poor predictors of the risk to exposure to HBV infection. CONCLUSION: The prevalence of HBV infection in heterosexual patients in the West Midlands is very low and does not provide any indications to broaden HBV vaccination into heterosexual patients attending GUM clinics. Risk factors were poor predictors of the exposure to HBV infection. This is partially due to the low prevalence of HBV infection in this study. Further studies are required before definitive conclusions are made regarding the potential predictive value of risk factors. PMID- 9215098 TI - Ectopic pearly penile papules: a paediatric case. PMID- 9215097 TI - Seropositivity against HPV 16 capsids: a better marker of past sexual behaviour than presence of HPV DNA. AB - OBJECTIVES: To assess if seropositivity to human papillomavirus type 16 capsids is a better marker of sexual history than the presence of HPV DNA. STUDY DESIGN: A population based age stratified random sample of 234 Norwegian women (mean age 32.8 years, range 20-44) was examined for HPV serum antibodies, cervical HPV DNA, cytology and age in relation to sexual behaviour. RESULTS: Neither age nor age at first sexual intercourse was associated with HPV 16 antibodies. Adjusted ORs for 4-5; 6-10 and > 10 versus 0-1 lifetime sexual partners, were 13.1 (95% CI 1.5 110.8), 8.2 (1.0-69.6) and 10.5 (1.2-94.0) for HPV 16 seropositivity, respectively; and 2.6 (0.2-27.8), 3.4 (0.4-31.7) and 4.1 (0.4-42.8) for HPV 16 DNA positivity, respectively. CONCLUSION: Seropositivity to HPV 16 capsids is positively associated with the number of sexual partners, suggesting that HPV 16 is predominantly sexually transmitted. The fact that serology had a stronger association with number of sexual partners than viral DNA suggests that seroreactivity is a better measure of lifetime history of HPV infection. PMID- 9215099 TI - Pearly penile papules. PMID- 9215100 TI - How to interpret an overview: a meta-analysis of the relative efficacy and toxicity of Pneumocystis carinii prophylactic regimens. PMID- 9215101 TI - Genitourinary medicine and the Internet No 5. PMID- 9215102 TI - Acquisition of pharyngeal gonorrhoea via sweets passed by mouth. PMID- 9215103 TI - An unusual presentation of vulvar carcinoma: a traumatic aetiology? PMID- 9215104 TI - Genital Chlamydia trachomatis infection in women in a Nigerian hospital. PMID- 9215105 TI - Provision of diagnostic services for genital chlamydial infection in genitourinary medicine clinics: England and Wales 1996. PMID- 9215106 TI - Same day testing for HIV: 1 year's experience in a district general hospital and at an alternative site. PMID- 9215107 TI - Survival and treatment of AIDS patients 1984-1993. PMID- 9215108 TI - Who goes to sexually transmitted diseases clinics? Results from a national population survey (Genitourin Med 1996; 72:197-202) PMID- 9215109 TI - Urinary symptoms, sexual intercourse and significant bacteriuria in male patients attending STD clinics. PMID- 9215110 TI - Antibiotic treatment for gonorrhoea in the UK. PMID- 9215111 TI - Epidemiology of gonococcal and chlamydial infections in Harrow and Brent. PMID- 9215112 TI - Epidemiological treatment and tests of cure in gonococcal infection: evidence for value. PMID- 9215114 TI - The Gulf War syndrome. PMID- 9215113 TI - Fetal and infant origins of adult disease. PMID- 9215115 TI - Apoptosis in haematological malignancies. PMID- 9215116 TI - Origins of ... image analysis in clinical pathology. PMID- 9215117 TI - Field study of lyophilised plasmas for local prothrombin time calibration in The Netherlands. AB - AIM: To assess the effect of a lyophilised calibrant plasma procedure on the international normalised ratio (INR) and its interlaboratory variation. METHODS: INR equivalent values were assigned to five lyophilised plasmas (one from normal donors and four from coumarin treated patients) by a reference laboratory using three calibrated thromboplastin reagents. The calibrant plasmas and five artificial control blood specimens were mailed to 44 Dutch laboratories for prothrombin time (PT) determination. The assigned INR values were used to calculate calibration lines for each participant laboratory. The calibration lines were then used to translate the PT of the control specimens to INR. RESULTS: For all lyophilised plasmas and control blood samples, there were significant differences between INR values determined with the three thromboplastin reagents. These differences could not be explained by inaccuracy of the international sensitivity index or mean normal PT of the reagents and must, therefore, have been induced by the preparation procedures for the lyophilised plasmas and control blood samples. The interlaboratory variation of the INR obtained with the calibrant plasma procedure had a coefficient of variation (CV) ranging between 2.1% and 7.3% and tended to be lower than the interlaboratory variation found with the usual methods (3.0-12.2% CV). There was a good agreement between the mean INRs obtained with the calibrant procedure and those obtained using the normal methods. CONCLUSIONS: The present study highlights the limitations of some lyophilised plasmas and control blood samples. It is not possible to assign a single INR value to each of these lyophilised plasmas and control specimens that is valid for all thromboplastin reagents. Nevertheless, by using reagent specific INR equivalent values for the calibrant plasma procedure, the interlaboratory variation could be reduced. PMID- 9215118 TI - Haematological validation of a computer-based bone marrow reporting system. AB - AIMS: To prove the safety and effectiveness of "Professor Belmonte", a knowledge based system for bone marrow reporting, a formal evaluation of the reports generated by the system was performed. METHODS: Three haematologists (a consultant, a senior registrar, and a junior registrar), none of whom were involved in the development of the software, compared the unedited reports generated by Professor Belmonte with the original bone marrow reports in 785 unselected cases. Each haematologist independently graded the quality of Belmonte's reports using one of four categories: (a) better than the original report (more informative, containing useful information missing in the original report); (b) equivalent to the original report; (c) satisfactory, but missing information that should have been included; and (d) unsatisfactory. RESULTS: The consultant graded 64 reports as more informative than the original, 687 as equivalent to the original, 32 as satisfactory, and two as unsatisfactory. The senior registrar considered 29 reports to be better than the original, 739 to be equivalent to the original, 15 to be satisfactory, and two to be unsatisfactory. The junior registrar found that 88 reports were better than the original, 681 were equivalent to the original, 14 were satisfactory, and two were unsatisfactory. Each judge found two different reports to be unsatisfactory according to their criteria. All 785 reports generated by the computer system received at least two scores of satisfactory or better. CONCLUSIONS: In this representative study, Professor Belmonte generated bone marrow reports that proved to be as accurate as the original reports in a large university hospital. The haematology knowledge contained within the system, the reasoning process, and the function of the software are safe and effective for assisting haematologists in generating high quality bone marrow reports. PMID- 9215119 TI - Girls with virilisation in childhood: a diagnostic protocol for investigation. AB - AIM: To analyse critically a protocol for the investigation of girls presenting with virilisation in childhood. METHODS: Twenty five girls aged 1.6-8.7 years with features of virilisation were evaluated. Twenty four had presented with pubic hair, eight with auxilliary hair, seven with facial acne, four with clitoromegaly, and 10 with tall stature. They underwent clinical assessment (height, weight, height velocity, staging of puberty, physical examination for acne, body odour, and clitoromegaly) and laboratory assessment comprising basal concentrations of cortisol, 17 OH-progesterone (17 OHP), androstenedione, dehydroepiandrosteronesulphate (DHEAS), testosterone, and oestradiol. The above steroids were also measured during the short synacthen test (0.25 mg intramuscularly) in 16 subjects and low dose dexamethasone suppression tests (0.5 mg at six hourly intervals over 48 hours). Pelvic ultrasound, computed tomography and magnetic resonance imaging of adrenals were carried out when the biochemical findings suggested that there might be an autonomous source of androgen secretion. RESULTS: Clinical and laboratory assessments differentiated the patients into three diagnostic categories: adrenarche (18 cases), congenital adrenal hyperplasia (five cases), and adrenocortical tumour (two cases). The last had elevated concentrations of DHEAS, 1.5 and 19.1 mumol/l (normal value < 0.5 mumol/l), androstenedione, 24.6 and 21.8 nmol/l (normal < 1 nmol/l), and testosterone, 4.5 and 2.4 nmol/l (normal < 0.8 nmol/l), with none suppressing on dexamethasone suppression. Congenital adrenal hyperplasia subjects had elevated basal serum concentrations of 17 OHP (n = 4): 250, 140, 14, and 14.1 nmol/l (normal < 10 nmol/l) and elevated peak values of 17 OHP after synacthen (n = 3): 76, 179.5, and 175 nmol/l. Adrenarche patients had elevated basal concentrations of DHEAS (median: 2.3 mumol/l; n = 17) and androstenedione (median 2.6 nmol/l; n = 17). Nine patients also had elevated basal serum testosterone concentrations (median 0.9 nmol/l). Peak values of 17 OHP after synacthen were significantly different from baseline (n = 12) and were < 50% of the lowest value in congenital adrenal hyperplasia. Serum DHEAS, androstenedione, and testosterone suppressed following dexamethasone suppression (n = 16), thereby distinguishing adrenarche patients from adrenal tumour patients. Clinical details did not distinguish patients, except for clitoromegaly which was present only in the tumour and congenital adrenal hyperplasia patients. CONCLUSIONS: This protocol proved useful and practical in cases of virilisation presenting particular diagnostic difficulty. PMID- 9215120 TI - Detection of apoptosis by the TUNEL technique in clinically localised prostatic cancer before and after combined endocrine therapy. AB - AIMS: Apoptosis in prostate cancer was evaluated after three months of combined endocrine therapy to investigate the association with tumour grade, tumour stage, and the immunohistochemical detection of p53 and bcl-2 in tumour cells before and after therapy. METHODS: Twenty six formalin fixed, paraffin wax embedded core biopsies and corresponding prostatectomy specimens, excised after three months of combined endocrine therapy, were analysed for the presence of apoptotic cells by the terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labelling (TUNEL) method, and for p53 and bcl-2 overexpression by immunohistochemistry. RESULTS: All 26 adenocarcinomas were clinically localised at diagnosis. In biopsies performed before combined endocrine therapy, the apoptotic indices varied between 0.09% and 1.73%, while the tumour grade fell between Gleason score 1 and 8. The mean (SD) apoptotic count pretherapy was 0.71% (0.50). There was a significant association between elevated apoptotic counts and higher Gleason scores in the biopsies (p = 0.005). After three months of therapy, the percentage of apoptotic tumour cells increased independently of tumour stage, while a significant association with Gleason grade was found (p = 0.0018) and all the tumours had Gleason scores of < 7. In eight cases the apoptotic index was more than twice its pretherapy value. The remaining tumours showed less of an increase in the apoptotic index (five cases) or a reduction in the percentage of apoptotic cells. The overall moderate increase in apoptotic index after combined endocrine therapy was not statistically significant (p = 0.8). Immunoreactivity to p53 was absent in all cases, before and after therapy, while a slight increase in the number of cells overexpressing bcl-2 was observed in five of the 13 tumours (38.1%) with reduced apoptotic indices after therapy. CONCLUSIONS: After three months of combined endocrine treatment a minority of clinically localised prostate neoplasms showed regressive epithelial alterations, associated with an increase in apoptotic tumour cells; an increase in cells overexpressing bcl-2 was observed in five of the 13 tumours with reduced apoptotic indices. PMID- 9215121 TI - An enhanced immunocytochemical method for staining bone marrow trephine sections. AB - AIMS: The detection of cellular antigens in fixed decalcified bone marrow trephine (BMT) sections depends on the method of processing, the nature of the antigen and antibody, antigen retrieval techniques, and the sensitivity of the immunocytochemical method. This study evaluated a tyramide enhanced avidin-biotin immunostaining method on formalin fixed decalcified BMT sections to determine whether the method could detect previously undetectable antigens. METHODS: Nineteen BMT biopsies from a range of haematological disorders were evaluated with 43 antibodies to haemopoietic antigens using horseradish peroxidase and alkaline phosphatase detection methods, using the tyramide enhanced avidin-biotin immunostaining method. RESULTS: Compared with standard avidin-biotin immunostaining methods the tyramide enhanced immunostaining method showed enhanced signal intensity, gave positive labelling for antigens that require pretreatment by other methods, and previously unreactive antigens were detected. Primary antibodies could be used at up to 200 times higher dilutions. CONCLUSION: The tyramide enhanced immunostaining method, while retaining specificity, is highly sensitive and enables an increased number and range of antigens to be detected than previously possible. The method could be applied to BMT sections for the routine diagnosis and classification of haematological disorders. PMID- 9215122 TI - Apoptosis in human hepatocellular carcinoma and in liver cell dysplasia is correlated with p53 protein immunoreactivity. AB - AIMS: To investigate the prevalence of apoptosis in human hepatocellular carcinomas (HCC) of different types and grades and in liver cell dysplasia, and to test whether the apoptotic rate is correlated with the p53 protein status. METHODS: 37 HCC and 66 six liver samples with liver cell dysplasia were analysed for apoptosis using in situ DNA end labelling (ISEL), and for p53 protein expression by immunohistochemistry. In HCCs, proliferative activity was quantitatively assessed using proliferating cell nuclear antigen labelling. RESULTS: The apoptotic index in HCC as based on ISEL ranged from 0.1 to 13.5 per 1000 cells analysed and was not related to type or grade. No nuclear staining was observed in multinuclear tumour cells. There was a significant correlation between the apoptotic rate and both the proliferative activity and p53 protein reactivity. In liver samples containing p53 protein positive liver cell dysplasia cells, there was a significantly higher apoptotic rate of these cells. CONCLUSIONS: Apoptosis is detectable in HCC, and is not related to type and grade. There is a highly significant positive correlation between the apoptotic rate in HCC and both the proliferative activity and p53 protein expression. A similar phenomenon occurs for putative cancer precursors. The findings support the role of p53 in regulating apoptosis in preneoplastic and neoplastic liver lesions. PMID- 9215123 TI - In situ detection of lipid peroxidation in chronic hepatitis C: correlation with pathological features. AB - AIMS: To assess the occurrence of lipid peroxidation in chronic hepatitis C and to evaluate its relation to pathological features and liver iron concentrations. METHODS: Liver biopsy samples of 43 patients with untreated chronic hepatitis C were studied by immunohistochemistry using specific antibodies directed against two major aldehyde metabolites of lipid peroxidation, malondialdehyde (MDA), and 4-hydroxynonenal (HNE). RESULTS: MDA and HNE adducts (aldehydes covalently linked to another molecule) were detected in the liver samples in 77% and 30% of cases, respectively. MDA adducts were detected both in the extracellular matrix and sinusoidal cells localised in areas of periportal and lobular necrosis. HNE adducts appeared in the cytoplasm of only a few hepatocytes. Comparison of the semiquantitative assessment of adducts (MDA and HNE indexes) with the grading and the staging of chronic hepatitis showed that the MDA index was correlated with fibrosis score (p < 0.001) and the grade of activity (p < 0.01). There was also a tendency to correlation with liver iron concentration (p = 0.09). No correlation was observed between the HNE index and pathological features or liver iron concentration. CONCLUSION: Lipid peroxidation products are detectable in the liver of chronic hepatitis C patients. The presence of MDA adducts in areas of active fibrogenesis and the correlation between the MDA index and fibrosis score suggest a role for lipid peroxidation in liver fibrosis. PMID- 9215124 TI - Analysis of phosphorylation of pRB and its regulatory proteins in breast cancer. AB - AIM: In order to study the role of retinoblastoma protein (pRB) in breast cancer, the phosphorylation of pRB and the expression of its related proteins-such as cyclin E, cyclin dependent kinase 2 (Cdk2), and p21/Cdk interacting protein 1 (Cip1)-were examined in 30 breast cancers in which pRB overexpression was confirmed immunohistochemically. METHODS: The phosphorylation of pRB for 30 tumours was investigated with western blotting. The expression of pRB, Cdk2/Cdc2, cyclin E, and p21/Cip1 was identified by immunohistochemistry and western blotting. RESULTS: The expression of pRB was confirmed in 52 of 70 tumours (74%) by immunostaining. Western blotting for pRB showed that 25 of 30 representative cancers (83%) were underphosphorylated, while only five tumours showed the hyperphosphorylated form of pRB. However, cyclin E and Cdk2-which promote phosphorylation of pRB-were expressed in all tumours. On the other hand p21/Cip1, a Cdk2 inhibitor, was expressed in 18 of 25 tumours with underphosphorylated pRB, while four of the five tumours with hyperphosphorylated pRB showed no expression of p21/Cip1. Examination of the relation between pRB phosphorylation and clinicopathological variables showed that the underphosphorylated group was characterised by low risk of lymph node metastasis (p < 0.01). CONCLUSIONS: The phosphorylation of pRB appears to be regulated mainly by p21/Cip1 through the suppression of cyclin E and Cdk2 in breast cancer. The underphosphorylated form of pRB may be useful as a prognostic factor. PMID- 9215125 TI - Relation between retinoblastoma and p53 proteins in human papilloma viruses 16/18 positive and negative cancers of the uterine cervix. AB - AIM: To ascertain the extent of retinoblastoma protein (pRB) expression in comparison to p53 protein and human papilloma viruses (HPV) 16/18 status in cervical carcinomas. METHODS: Fifty cases of invasive cervical carcinoma were HPV typed for genotypes 16 and 18 using consensus primers by polymerase chain reaction (PCR). Immunohistochemistry for pRB and p53 was done on formalin fixed tissue using microwave antigen retrieval and commercially available antibodies. RESULTS: Forty five cases were squamous carcinomas, three were adenocarcinomas, and two were adenosquamous carcinomas. Thirty one cases were HPV 16 positive and one was HPV 18. Sixteen cases showed +4 pRB expression and a further 11 were +3 positive. Seven cases were negative. Only five cases (10%) showed +4 p53 immunostaining, while seven were negative and 15 were +1. Of the 16 pRB +4 positive cases, one was negative for p53 and a further seven were +1 positive. This inverse pattern of staining between pRB and p53 had a p value of < 0.001. No correlation was observed between HPV 16/18 status and p53 and/or pRB staining. CONCLUSIONS: pRB is expressed in the majority of cases of cervical cancer (86%), with more than 75% (+4) of the tumour cell population being positive in 16 cases (32%). There appears to be a general inverse pattern of staining between pRB (high) and p53 (low) in cervical cancer. The expression of both pRB and p53 proteins is independent of the HPV 16/18 status of the tumour. PMID- 9215127 TI - Tissue preparation for immunocytochemistry. AB - AIMS: To investigate the effect of tissue preparation on immunostaining and to establish whether there is a standard tissue preparation schedule that allows optimal demonstration of all antigens. METHODS: Blocks of tonsil were subjected to variations to a standard fixation, processing, and section preparation schedule. The sections were stained with five antibodies-L26 (CD20), UCHL1 (CD45RO), CD3, vimentin, and anti-kappa light chain--using the streptavidinbiotin immunostaining technique. When further investigation was necessary, other tissues and antibodies were used and where weak immunostaining was obtained the use of microwave pretreatment to improve staining was tested. RESULTS: Several factors involved in fixation were found to affect immunoreactivity. These included the duration, pH, and type of fixative used. In tissue processing only temperature and the duration of the dehydration and wax infiltration steps affected immunoreactivity. Of all the factors investigated, the temperature and duration of the section drying had the greatest effect. In contrast, long term storage of cut sections before immunostaining had no effect on the reactivity of the antibodies tested. Antibodies were found to be affected by alterations to tissue preparation by varying degrees, UCHL1 and vimentin being the most susceptible to changes in fixation and L26 to changes in processing. Where weak staining occurred, microwave pretreatment was generally found to eliminate the problem. CONCLUSIONS: There is no standard tissue preparation schedule for the optimal demonstration of all antigens. Factors involved in all aspects of tissue preparation can affect immunoreactivity, so it is important that precise details of the preparation schedule are given when reporting immunocytochemical studies, rather than using the general term "routinely fixed and processed". PMID- 9215126 TI - MUC1 and MUC2 mucins in flat and polypoid colorectal adenomas. AB - AIMS: To examine the expression of MUC1 and MUC2 apomucins and distribution of MUC phenotypes (MUC2+/ MUC-, MUC2+/MUC1+, MUC2-/ MUC1+, MUC2-/MUC1-) in colorectal tubular adenomas in order to compare the distribution of phenotypes in flat and polypoid adenomas. METHODS: Endoscopically resected specimens of 35 flat and 15 polypoid tubular adenomas measuring less than 10 mm were examined and compared for the expression of MUC1 (MUSE11) and MUC2 (CCP58) and combined MUC phenotype distribution using conventional immunohistochemistry. RESULTS: There was no significant difference between flat and polypoid adenomas in their expression of MUC1 and MUC2 and the MUC phenotype distribution when stratified by grade of histological atypia. Adenomas with low grade atypia showed more extensive MUC2 expression than MUC1 (MUC2+/MUC1-phenotype). Expression of MUC1 was more extensive in adenomas with high grade atypia and the majority displayed either MUC2+/ MUC1+ or MUC2-/MUC1+ phenotypes. CONCLUSIONS: MUC2/MUC1 phenotypes were similar in flat and polypoid adenomas when stratified by grade of atypia. High grade atypia was characterised by reduced MUC2 and increased MUC1 expression in both types of adenoma. The phenotype MUC2-/MUC1+ occurs in tubular adenomas and cannot be a specific marker for de novo colorectal cancer. PMID- 9215128 TI - Sporicidal activity of chemical and physical tissue fixation methods. AB - AIMS: The effects of alcohol based fixation and microwave stimulated alcohol fixation were investigated on spores of Bacillus stearothermophilus and Bacillus subtilis (var. niger). METHODS: Spores were exposed to 10% formalin, or different concentrations of various alcohol containing fixatives (Kryofix/Spuitfix). Adequate controls were also set up in conjunction with the test solutions. The spores were immersed with and without adjunctive microwave stimulation in the various solutions tested. Possible surviving spores were recovered in revival broth and after incubation, and Gram staining viable counts were performed. RESULTS: Alcohol based fixatives did not have a sporicidal effect on B stearothermophilus or B subtilis (var. niger) spores, and microwave stimulated alcohol fixation at 450 W and up to 75 degrees C did not have a sporicidal effect. CONCLUSIONS: When alcohol based fixatives are used for fixation, precautions should be taken with the material thus treated, as it may contain viable spores or other pathogens, which are destroyed after 24 hours of formalin treatment. Of the physicochemical methods tested involving microwaving, none was successful in eliminating viable spores from the test material. PMID- 9215129 TI - Haemoglobin O Padova and falsely low haemoglobin A1c in a patient with type I diabetes. AB - Glycated haemoglobin (HbA1c) measured by high performance liquid chromatography (HPLC) in a 20 year old female with insulin dependent diabetes mellitus was consistently within the normal range although her daily blood glucose values were > 11.1 mmol/l. HbA1c measured by immunoagglutination and fructosamine was elevated and correlated with the patient's blood glucose values. The HPLC chromatogram showed an additional peak at HbA0. Electrophoresis of haemoglobin on citrate agar gel revealed an abnormal haemoglobin anodal of HbS. Cellulose acetate electrophoresis and isoelectric focusing demonstrated an additional haemoglobin migrating close to HbA2. Amino acid analysis and DNA sequencing revealed an alpha 30 (B11) Glu-->Lys replacement, that is, haemoglobin O Padova. Investigations of two family members without diabetes revealed the same rare haemoglobin variant. This case showed that this silent haemoglobin mutation caused an additional peak and falsely low HbA1c values when measured by HPLC, the gold standard for this evaluation. PMID- 9215130 TI - Fatal Campylobacter jejuni infection in a patient splenectomised for thalassaemia. AB - A 35 year old man with a fatal Campylobacter jejuni infection is described. He had HbE/beta zero thalassaemia and had undergone splenectomy nine months previously for hypersplenism; he also had chronic hepatitis C infection. He presented with high grade fever but no gastrointestinal symptoms and rapidly progressed to septicaemic shock and hepatic encephalopathy despite treatment with penicillin, gentamicin, and, later, chloramphenicol and ceftazidime. Only one case of Campylobacter jejuni septicaemia occurring post-splenectomy has been reported previously, also in an iron overloaded thalassaemia patient. Unusual Gram negative bacilli must be covered by the chosen antibiotic regimen when splenectomised thalassaemic patients present with high grade fever. PMID- 9215132 TI - Widespread neuroendocrine malignancy within the central nervous system: a diagnostic conundrum. AB - A 75 year old female presented with a sellar tumour, and was subsequently found also to have a cauda equina tumour, a parietal dural tumour, a pontine tumour, an intradural spinal tumour, and several vertebral body tumours. Histological examination revealed a neuroendocrine tumour forming cell nests surrounded by reticulin. There was moderate nuclear pleomorphism, prominent mitoses, and focal necrosis. Immunohistochemistry showed diffuse positive staining with cytokeratins, chromogranin and 5-hydroxytryptamine, and focal positive staining with S100. This case is an unusual and ultimately insoluble, diagnostic problem; however, the differential diagnoses include pituitary carcinoma, malignant paraganglioma, and atypical carcinoid. PMID- 9215131 TI - Standardisation of polymerase chain reaction for the detection of Salmonella typhi in typhoid fever. AB - To improve the diagnosis of Salmonella typhi infection, a polymerase chain reaction (PCR) assay was developed for the amplification of the dH flagellin gene of S typhi. Primers were designed from dH flagellin gene sequence which will give an amplification product of 486 base pairs. In tests to study the specificity of the assay, no amplification was seen in non-salmonella strains or salmonella strains with flagellar gene other than "d". Sensitivity tests determined that 28 pg of S typhi target DNA or 3 x 10(2) target bacteria could be detected by the PCR assay. Subsequently, the PCR technique was used for detection of S typhi in blood or clot cultures from 84 patients clinically suspected of having typhoid fever, and from 20 healthy control subjects. Twenty five of 84 samples from clinically suspected cases were positive by PCR; four of which were culture negative. No amplification was seen in samples from patients who were culture positive for organisms other than S typhi or from controls. The time taken for each sample for PCR analysis was less than 48 hours compared with three to five days for blood or clot culture. PCR appeared to be a promising diagnostic test for typhoid fever. PMID- 9215133 TI - Effusion cytology of hepatocellular carcinoma with in situ hybridisation for human albumin. AB - While the cytological features of hepatocellular carcinoma on fine needle aspiration cytology are well described, cases of hepatocellular carcinoma with malignant cells in ascitic fluid and their characteristics are not. A patient is described with cirrhosis resulting from chronic hepatitis B virus infection, ascites, and hepatocellular carcinoma diagnosed by effusion cytology. The malignant cells in the effusion were shown to be positive for alpha fetoprotein using immunocytochemistry, and for human albumin using in situ hybridisation, confirming the diagnosis of hepatocellular carcinoma. Further investigations in a terminally ill patient were thus avoided. PMID- 9215134 TI - Dilutional hyponatraemia: a cause of massive fatal intraoperative cerebral oedema in a child undergoing renal transplantation. AB - A four year old boy with polyuric renal failure resulting from recurrent urinary tract infections and vesicoureteric reflux from birth underwent renal transplantation. In the past he had had five ureteric reimplant operations and a gastrostomy, as he ate nothing by mouth. He required peritoneal dialysis 13 hours a night, six nights a week. His fluid requirements were 2100 ml per day. This included a night feed of 1.5 litres Nutrizon. Before operation he received 900 ml of Dioralyte instead of the Nutrizon feed, and peritoneal dialysis was performed as usual. The operation itself was technically difficult and there was more blood loss than anticipated, requiring intravenous fluids and blood. The operation ended about four hours later but he did not wake up. Urgent computed tomography revealed gross cerebral oedema. He died the next day. At necropsy the brain was massively oedematous and weighed 1680 g. PMID- 9215135 TI - Biopsy diagnosis of prostatic cancer--current areas of concern. PMID- 9215136 TI - Colorectal cancer reporting: are we failing the patient? PMID- 9215137 TI - Is lipid oxidation essential for atherogenesis? PMID- 9215138 TI - Antioxidants, cholesterol, and ischaemic heart disease: CHAOS or confusion? PMID- 9215139 TI - What's new in the diagnosis of head injury? AB - The diagnosis of DAI is not always easy, and should be based on adequate sampling of appropriate anatomical areas from a sliced, fixed brain. It is now recognised that there is a continuum of traumatic white matter damage, and that DAI represents only the severe end of the scale. Such damage may be detected from very shortly after a head injury-a fact that may give rise to some challenging diagnostic problems. Early axonal injury detected by means of beta APP immunostaining should be interpreted with caution. The most useful tools currently available for detecting axonal damage are antisera to beta APP, PG-M1, and GFAP, used in conjunction with a routine haematoxylin and eosin stain, but even with immunocytochemistry precise dating of histological changes may not be possible. PMID- 9215140 TI - Origins of ... flow cytometry and applications in oncology. PMID- 9215141 TI - Evaluation of a novel endoluminal brush method for in situ diagnosis of catheter related sepsis. AB - AIMS: To determine the accuracy of a novel endoluminal brush method for the diagnosis of catheter related sepsis (CRS), which is performed in situ and hence does not require line sacrifice. METHODS: 230 central venous catheters in 216 patients were examined prospectively for evidence of CRS or colonisation using an endoluminal brush method in conjunction with peripheral blood cultures. The results were compared with those obtained using methods that require line sacrifice: extraluminal sampling (Maki roll) or endoluminal sampling (modified Cleri flush) of microorganisms. RESULTS: Only 16% of 128 patients suspected clinically of having line associated infection were confirmed as having CRS. In addition, 2 of 102 patients not suspected of having line associated infection had CRS. Line colonisation was apparent in approximately twice as many catheters using the Maki roll criteria (92%) compared with either the endoluminal brush (43%) or Cleri flush (43%). Furthermore, colonised catheters sampled using the Maki roll technique yielded mixed growth twice as often as when examined by endoluminal methods (17 and 8 cases, respectively). It was rare to detect either only endoluminal (4 of 22 episodes) or extraluminal (1 of 22 episodes) microorganisms in cases of CRS. In contrast, catheters defined as being colonised most frequently (59% of episodes) yielded only significant extraluminal growth. Only one case of CRS (5%) would have been "missed" if lines yielding a negative result from endoluminal brush sampling had been left in situ. Conversely, four episodes of CRS (18%) would not have been diagnosed by relying on extraluminal sampling alone. CONCLUSIONS: Diagnosis of CRS by the endoluminal brush method can be achieved without line sacrifice and is more sensitive (95%) and specific (84%) than extraluminal sampling of the catheter tip by the Maki roll technique (82% and 66%, respectively). PMID- 9215142 TI - Lessons for the laboratory from a general practitioner survey. AB - AIMS: To assess the current performance of the clinical biochemistry service provided to general practitioners, with particular attention to result turnround times, and to identify and improvements required. METHODS: Postal questionnaire survey of general practitioners in the London Borough of Tower Hamlets who used the clinical biochemistry laboratory of the Royal London Hospital. A flow analysis study of turnround times for general practitioner samples was also performed. RESULTS: Responses to the questionnaire showed that although 82% of general practitioners thought the current quality of service provided was better than fair, the actual turnround times achieved were longer than the acceptable times required. There was also a strong demand (> 66% of responders) for additional information-such as highlighting of abnormal results-to be provided with results. There was wide variability between practitioners in their use of the laboratory (from none to > 800 requests per year), with no apparent correlation to practice size. Of the repertoire of tests requested, a surprisingly high percentage (14.3%) were for thyroid function. Flow analysis of turnround times for thyroid function tests showed that problems lay not with the time taken for analysis (only 7.8% of the total turnround time) but with the pre- and postanalytical phases, that is, the sample collection and results delivery service. CONCLUSIONS: Increasing the proportion of health care delivered in the primary care sector will inevitably increase the requirement for pathology services. Improvements in the specimen collection and results delivery service to general practitioners are needed to meet their expectations. It remains to be determined whether increased investment in these aspects of laboratory service would result in improved patient care in the primary sector. PMID- 9215143 TI - Genetic analysis of hydatidiform moles in paraffin wax embedded tissue using rapid, sequence specific PCR-based HLA class II typing. AB - AIMS: To determine the applicability of rapid, sequence specific polymerase chain reaction (PCR)-based HLA class II genotyping for the distinction of complete from partial hydatidiform moles (HM) using DNA extracted from formalin fixed and paraffin wax embedded tissue. METHODS: Nine HM were studied. DNA was extracted from formalin fixed and paraffin wax embedded tissue after mechanical separation of decidual and molar components. HLA class II DRB (DRB1, -3, -4, and -5) and DQB1 genotyping was performed using a parallel series of PCR reactions, each of which contained sequence specific primers designed to amplify different HLA DRB and DQB1 alleles or allele groups (PCR-SSP analysis). In each case the HLA DRB and DQB1 genotypes identified within the decidua and HM were compared. RESULTS: Within the decidual tissue, HLA DRB genotypes were assignable in all nine cases, and HLA DQB1 genotypes were identified in seven cases. Within the molar tissue, HLA DRB genotypes were assignable in seven cases, and at least one HLA DQB1 allele was identified in seven cases. Interpretation based on HLA class II genotyping was therefore possible in two cases classified on histological appearances as complete HM, in four classified as partial HM, and in one HM of uncertain type. Different HLA DRB and DQB1 haplotypes were identified within the decidual and molar tissue from both complete HM, consistent with a solely paternal origin and supporting the histological diagnosis. HLA DRB and DQB1 alleles common to the decidual and molar tissue were present within the four partial HM and the HM of histologically uncertain type, consistent with combined maternal and paternal genetic input to these HM, supporting the histological diagnosis in four cases and suggesting that the histologically equivocal case was also a partial HM. CONCLUSION: PCR-SSP HLA class II DRB and DQB1 typing is reliably applicable to DNA extracted from formalin fixed and paraffin wax embedded tissue. Therefore, in a suitably equipped HLA typing laboratory, this technique provides a useful adjunct to histological examination for differentiation of complete from partial HM. PMID- 9215145 TI - Detection and comparative analysis of persistent measles virus infection in Crohn's disease by immunogold electron microscopy. AB - AIMS: To determine the specificity of persistent measles virus infection in intestinal samples from Crohn's disease patients using quantitative immunogold electron microscopy. To compare the results with samples from ulcerative colitis, a granulomatous inflammatory control (tuberculous lymphadenitis), and a positive control. METHODS: Formalin fixed, paraffin embedded intestinal tissue from patients with Crohn's disease was reprocessed and stained with antimeasles nucleocaspid protein primary antibody followed by 10 nm gold conjugated secondary antibody. Tissue samples were taken from granulomatous and non-granulomatous areas of the intestine. Intestinal samples from patients with ulcerative colitis, tuberculous lymphadenitis, or acute mesenteric ischaemia were similarly processed. Brain tissue from a patient with subacute sclerosing panencephalitis (SSPE) was used as the positive control. Duplicate sections of all tissues were processed without the primary antibody. Stained specimens were examined by electron microscopy. RESULTS: In Crohn's disease patients, 8/9 foci of granulomatous inflammation and 0/4 foci of non-specific inflammation were positive for measles virus. Of controls, 0/5 non-inflamed intestinal tissues, 1/8 tuberculous tissues, 1/5 ulcerative colitis tissues, and 1/1 SSPE tissues were positive. Gold grain counts per nuclear field-of-view in both Crohn's disease granulomas (43.29) and SSPE (36.94) were significantly higher than in tissues from patients with ulcerative colitis (13.52) or tuberculous lymphadenitis (15.875), and nongranulomatous areas of Crohn's disease (4.89) (p < 0.001, p < 0.001, p = 0.0006, respectively), with no significant difference between Crohn's disease and SSPE (p > 0.1). In both SSPE and Crohn's disease staining was confined to a small population of cells exhibiting characteristic cytopathology. CONCLUSION: These data support a role for measles virus in the aetiology of Crohn's disease. PMID- 9215144 TI - Intra-abdominal sepsis: an immunocytochemical study of the small intestine mucosa. AB - AIM: To investigate immunocytochemical changes in intestinal tissues from patients with intra-abdominal sepsis, and to relate the changes to the possibility of enhanced bacterial adhesion and translocation. METHODS: Tissues from 17 patients suffering from intra-abdominal sepsis and from controls were sectioned and stained immunocytochemically for IgA, IgM, secretory component, J chain, and HLA-DR. Differences in the distribution and characteristics of positively staining cells between the patient groups were assessed. RESULTS: Patients with intra-abdominal sepsis had noticeable reductions in numbers of IgA and IgM plasma cells, reduced J chain staining, and had little immunoglobulin on the surfaces of enterocytes. In contrast, HLA-DR positive cells were increased in the sepsis compared with the control group. The plasma cells present showed cytological changes suggestive of apoptosis. CONCLUSIONS: Stress associated with sepsis and its immediate causes might result in increased plasma glucocorticoid levels that bring about apoptosis of mucosal plasma cells (or their precursors). The consequent reduction in expression of IgA and IgM may favour bacterial adhesion to the enterocytes and facilitate bacterial translocation into the tissues. PMID- 9215146 TI - Langerhans' cells are depleted in chronic graft versus host disease. AB - AIMS: To measure Langerhans' cells in skin of patients treated by bone marrow transplantation who developed chronic graft versus host disease (GvHD); to determine whether the reduction in Langerhans' cells resulted directly from the GvHD or from other factors, such as the immunosuppressive regimens used in bone marrow transplant patients. PATIENTS AND METHODS: Lesional and nonlesional skin specimens from nine patients with lichen planus-like lesions and three patients with sclerodermoid lesions were studied. Control skin specimens were taken from three patients undergoing breast reduction surgery. The number of Langerhans' cells/mm2 and the area of Langerhans' cells as a percentage of total epidermis were measured by counting cells labelled with antihuman CD1a. RESULTS: A significant reduction in Langerhans' cell area and number were found in specimens with lesions (area 3.5%; number 507/mm2) compared with specimens without lesions (8.42%; 2375/mm2). In contrast, Langerhans' cell area and number in skin without lesions were similar to controls (10.26%; 2968/mm2). CONCLUSIONS: Langerhans' cells were significantly reduced in skin with lesions of chronic GvHD but not in skin without lesions from the same patient, suggesting that the reduction is a direct consequence of GvHD and not linked to immunosuppressive drugs or late effects of conditioning regimens. In long term bone marrow transplant recipients, Langerhans' cells are derived mainly from the donor cells; therefore, this result suggests the occurrence of autoreactive phenomenon in chronic GvHD. PMID- 9215147 TI - Detection of perforin and tumour necrosis factor alpha mRNA expressing cells in sclerosing lymphocytic lobulitis of the breast. AB - The contribution of a cellular immune response to tissue destruction in sclerosing lymphocytic lobulitis of the breast is not well understood. In this study, comparison of one case with two age matched control cases showed an increased frequency of activated perforin mRNA expressing cells at the site of tissue destruction in lobulitis. Along with the detection of tumour necrosis factor alpha (TNF alpha) mRNA expressing cells in the infiltrates, the striking association of perforin expressing activated cytotoxic cells with remaining gland parenchyma and the high level of perforin mRNA suggests activation of cytotoxic cells in situ. These findings are evidence that cell mediated cytotoxicity plays a significant role in the destruction of mammary gland tissue in sclerosing lymphocytic lobulitis. PMID- 9215149 TI - p53 gene product expression in resected non-small cell carcinoma of the lung, with studies of concurrent cytological preparations and microwave antigen retrieval. AB - AIM: To document the frequency and extent of p53 gene product expression in paraffin sections of resected non-small cell carcinoma of the lung and in cytological preparations of the same tumours; to determine the effect of microwave antigen retrieval on antigen detection. METHODS: Representative paraffin sections of 50 non-small cell carcinomas were stained with an antibody to p53 gene product (DO-7) both with and without prior microwave antigen retrieval. Cytoblocks and cell smears obtained from 19 cases were similarly stained. RESULTS: Using a histochemical scoring system (0-300) which takes into account staining intensity and extent, 78% (n = 39) of microwave pretreated paraffin sections and 52% (n = 26) of non-pretreated sections scored between 5 and 300; p = 0.001; 56% (n = 28) of microwave pretreated sections and only 2% (n = 1) of non-pretreated sections scored between 100 and 300 (p = 0.0001); 75% of direct smears of tumours and 80% of cytoblocks stained similarly to the paraffin sections of the resected specimens. No smears or cytoblocks stained positively when the sections of the resected specimen were negative. CONCLUSIONS: As up to 78% of non-small cell lung carcinomas overexpress p53 gene product, this may prove to be a valuable diagnostic method in biopsy or cytological material when the morphological diagnosis is uncertain. Microwave antigen retrieval is effective on formalin fixed tissue. PMID- 9215148 TI - Differential expression of the urokinase receptor (CD87) in arthritic and normal synovial tissues. AB - AIM: To determine whether the urokinase plasminogen activator receptor (u-PAR; CD87) exhibits a possible pathogenic role in rheumatoid and osteoarthritis. METHODS: A semiquantitative, indirect immunoperoxidase histochemical analysis was performed on frozen synovial tissue sections. The recently characterised monoclonal antibody 10G7 recognising transfectants bearing u-PAR was used. Synovial tissue was obtained from 10 patients with rheumatoid arthritis, 10 patients with osteoarthritis, and four normal subjects. RESULTS: u-PAR was expressed on 70-90% of synovial tissue lining cells and subsynovial, interstitial macrophages from the arthritis patients, but only on a few myeloid cells from the normal subjects. It was also present on more endothelial cells from the rheumatoid and osteoarthritis patients, than from normal synovial tissue. CONCLUSIONS: Plasminogen activators are important in joint destruction underlying arthritis. The up-regulated expression of u-PAR in diseased versus normal synovial tissue suggests a role for this antigen in the inflammatory and angiogenic mechanisms underlying rheumatoid and osteoarthritis. PMID- 9215150 TI - Ovarian granulomas: a report of 32 cases. AB - AIMS: To determine the causes of ovarian granulomatous inflammation and to discuss the differential diagnoses. METHODS: The pathological features of all ovarian granulomas identified by pathology SNOMED coding in Northern Ireland over a 13 year period were reviewed. Case notes of patients were also reviewed. RESULTS: The most common cause of ovarian granuloma was a foreign body reaction to suture material introduced at a previous operative procedure (15 cases). Other causes were Crohn's disease (four cases), previous diathermy (two cases), tuberculosis (two cases), a necrotising reaction following previous surgery (two cases), endometriosis (one case), and bacterial tubo-ovarian abscess (one case). In five cases, no cause was apparent for the granulomatous inflammation. In these, varying numbers of small, well circumscribed cortical granulomas were present. These cases correspond to so-called "idiopathic" cortical granulomas. CONCLUSION: The study confirms the wide range of conditions which can give rise to ovarian granulomatous inflammation. PMID- 9215151 TI - The hormonal receptor status of uterine carcinosarcomas (mixed mullerian tumours): an immunohistochemical study. AB - AIM: To investigate the role of oestrogen and progesterone receptor status in uterine carcinosarcomas (mixed Mullerian tumours) to see whether the receptors were identifiable, and if so whether they were of significance clinically. METHODS: 11 cases of uterine carcinosarcoma were identified from clinical and pathology records. An immunohistochemical method was used to demonstrate oestrogen and progesterone hormone receptors on paraffin embedded material, with suitable tissue controls, staining being recorded. RESULTS: 10 of 11 cases showed staining for one or both hormone receptors in normal tissue adjacent to tumour. In four carcinosarcoma cases, staining for one or both receptors was shown within the epithelial component (appearing to correlate with the degree of epithelial differentiation); two of these cases had staining within sarcomatous areas. Two of the three patients still alive had epithelial hormone receptor positivity. CONCLUSIONS: Receptors for oestrogen and progesterone were found in four of 11 cases of uterine carcinosarcoma, using paraffin embedded material. There may be an association between hormone receptor positivity and clinical outcome. PMID- 9215152 TI - Lancefield grouping and smell of caramel for presumptive identification and assessment of pathogenicity in the Streptococcus milleri group. AB - AIM: To evaluate Lancefield grouping and caramel smell for presumptive identification of the Streptococcus milleri group, and to find whether Lancefield group, species, or protein profile correlated with virulence or infection site. METHODS: Prospective studies were made of 100 consecutive streptococcal isolates in blood cultures or pus from 100 patients in whom the severity of infection was categorised as serious, moderate, or not significant. The usefulness of Lancefield group and the caramel smell for presumptive identification was examined, and the relation of the S milleri species, Lancefield group, and SDS PAGE protein analysis to severity of infection and infection site was investigated. Lower respiratory tract and genital tract specimens, strict anaerobes, group D streptococci, and strains identified as Streptococcus pneumoniae, Streptococcus pyogenes, or Streptococcus agalactiae were excluded. RESULTS: Most streptococci occurring in pure or significant growth density were S milleri group (87/100; 87%, 95% confidence interval 0.81-0.93). Of these, 89.7% (78/87; 0.84-0.96) were associated with infection. Lancefield group F antigen predominated (41/87; 47.1%, 0.38-0.56). Lancefield group F alone or accompanied by the caramel smell had a specificity of 100%, but a sensitivity of only 47.3% for group F alone, and 19.5% for group F accompanied by the caramel smell. There was no significant association between species, Lancefield group, and severity of infection, site of infection, or pathogenicity. SDS-PAGE analysis failed to discriminate between strains. CONCLUSIONS: Neither species nor Lancefield antigen was related to the site of infection. The presence of Lancefield group F antigen alone or accompanied by a caramel smell was a useful indicator for the S milleri group when present, but was too insensitive to use as a screening test. Most streptococci occurring in pure culture or in significant growth density were of clinical importance. Such organisms should be identified to species level to detect the S milleri group. PMID- 9215153 TI - Urinary tissue factor in glomerulonephritis: a potential marker of glomerular injury? AB - AIM: To investigate the significance of urinary tissue factor (uTF) concentrations in patients with glomerulonephritis. METHODS: Urine samples were collected from normal subjects (n = 57), patients with uncomplicated renal stones (n = 30), and patients with glomerulonephritis (n = 150). Samples were then centrifuged and the pellets solubilised in n-octyl-beta-glucopyranoside. uTF concentrations were determined using a one stage kinetic chromogenic assay. RESULTS: The uTF concentration was higher in patients with glomerulonephritis than in normal controls (p < 0.01) or in patients with renal stones (p < 0.05). uTF activity correlated with the protein creatinine index (PCI, r = 0.41, p < 0.001) and seven patients with glomerulonephritis and a PCI < or = 0.1 g/mmol had raised uTF. Glomerulonephritis patients were subdivided into two groups depending on the PCI: < 0.2 g/mmol creatinine (mild to moderate proteinuria, group I) and > or = 0.2 g/mmol creatinine (heavy proteinuria, group II). In group I, uTF concentrations were higher in patients with either immune complex (IC) glomerulonephritis (p < 0.01) or non-IC (p < 0.05) glomerulonephritis than in normal controls. In group II, the IC glomerulonephritis group had higher uTF concentrations than normal controls (p < 0.001) or patients with renal stones (p < 0.01); and non-IC glomerulonephritis patients had higher uTF than normal controls (p < 0.01). When the glomerulonephritis groups were divided into broad WHO subtypes, the significance level varied with the type of glomerulonephritis. CONCLUSIONS: uTF is increased in patients with glomerulonephritis, and its concentration may reflect the aetiopathogenesis of glomerulonephritis. PMID- 9215155 TI - Automated type specific ELISA probe detection of amplified NS3 gene products of dengue viruses. AB - AIM: To apply an automated system of nucleic acid hybridisation coupled with the enzyme linked immunosorbent assay (ELISA) for the type specific detection of amplification products of dengue viruses. METHODS: Non-structural 3 (NS3) gene targets of reference strains of all four dengue and other flaviviruses, as well as dengue patient viraemic sera, were subjected to reverse transcription and polymerase chain reaction using consensus and dengue type specific primers and digoxigenin-11-dUTP label incorporation. The amplification products were detected by biotinylated type specific primers which served as ELISA capture probes bound to streptavidin coated tubes. RESULTS: Significantly high spectrophotometric absorbance readings were obtained by hybridisation of the consensus and seminested amplification products of all four dengue viruses with their respective capture probes. In contrast, extremely low absorbances were observed for consensus products of Japanese encephalitis, yellow fever, and Kunjin viruses, which served as negative controls. These ELISA data correlated well with agarose gel electrophoresis of dengue type specific amplified products of diagnostic sizes. CONCLUSIONS: The combination of in vitro amplification and antibody based detection offers rapid, type specific, high throughput, and gel free detection of amplified products of dengue viruses. PMID- 9215154 TI - Effect of interferon alpha on high serum androgen concentrations in HIV positive men with Kaposi's sarcoma. AB - AIM: To measure serum androgen concentrations in men with HIV related Kaposi's sarcoma (KS) who had been treated with recombinant interferon (IFN) alpha-2a to determine the role of androgens on the development of KS lesions. METHODS: 32 men with HIV related KS who had been treated with IFN were studied: 24 men in complete KS remission and eight not in remission. Serum androgen concentrations were determined before, during, and after IFN treatment and correlated with clinical remission. RESULTS: All patients in complete KS remission had lower serum androgen concentrations following IFN treatment: -51% for dehydroepiandrosterone (DHEA) (p < 0.0001); -38% for DHEA sulphate (p < 0.002); 39% for androstenedione (p < 0.002); and -44% for testosterone (p < 0.007). These decreases brought the serum concentrations to about normal levels. However, IFN had varying effects on serum androgen concentrations in the men not in remission: a small decrease, a large increase in one androgen, or no change in serum androgens. CONCLUSIONS: The association between serum androgen levels and the progression or remission of HIV associated KS suggests that androgens affect the development of KS lesions. A clear understanding of the changes in the androgen environment may provide a sound basis for the development of new therapeutic strategies. PMID- 9215156 TI - Late stage congestive gastropathy. AB - Gastric mucosal abnormalities resulting from portal hypertension are defined as "congestive gastropathy". A case of congestive gastropathy with unusual features, in a 63 year old man with a history of excessive alcohol intake and cirrhosis, is described. The patient underwent a subtotal gastrectomy because of profuse bleeding from a gastric ulcer, providing a large surgical specimen for examination. Unusual gross and histological findings included prominent arterial intimal hyperplasia, and diffuse duplication and focal fragmentation of the internal lamina elastica. The differential diagnosis of this condition includes primary angiodysplastic gastropathy such as Dieulafoy's disease. The similarity with Dieulafoy-like angiodysplasia emphasises that clear cut criteria to define gastric vascular lesions do not yet exist. PMID- 9215157 TI - Inflammatory pseudotumour of the liver. AB - Inflammatory pseudotumour is not a common lesion. The first series of 12 cases was described in 1986, to which 37 more cases have now been added. The histology, differential diagnosis, and prognosis of this lesion have been described in detail, but the aetiology is unknown and the mode of treatment remains controversial. A new case is presented and compared with the previously reported cases. Fine needle aspirate yielded a growth of klebsiella organisms. The possibility of this infection as an aetiological agent is considered. PMID- 9215158 TI - Benefits and limitations of pathology databases to cancer registries. PMID- 9215160 TI - Rights of possession in human corpses. PMID- 9215159 TI - Histological patchiness and sparing of the rectum in ulcerative colitis: refuting the dogma. PMID- 9215161 TI - Proliferation indexes--a comparison between cutaneous basal and squamous cell carcinomas. PMID- 9215162 TI - Brain tissue banks in psychiatric and neurological research. PMID- 9215163 TI - Laboratory diagnosis of malaria. PMID- 9215164 TI - Managed care: a ball control game. PMID- 9215165 TI - Intrapartum oligohydramnios does not predict adverse peripartum outcome among high-risk parturients. AB - OBJECTIVE: Oligohydramnios can be defined by an amniotic fluid index < 5th percentile for gestational age or an amniotic fluid index < or = 5.0 cm regardless of gestational age. The purpose of this prospective study was to determine whether oligohydramnios by either definition predicts accurately, in a high-risk population, the risks for cesarean section for fetal distress, Apgar score < 7 at 5 minutes, and neonatal acidosis. STUDY DESIGN: An amniotic fluid index was obtained in 490 consecutive parturients with medical or obstetric complications and a reliable gestational age. After each delivery, an umbilical arterial blood gas analysis was obtained. Both measures of amniotic fluid index were rated as screening tests with use of sensitivity, specificity, predictive values, and receiver-operator characteristic curves. RESULTS: The incidences of cesarean section for fetal distress and umbilical arterial pH < 7.00 were 14% and 1.8%, respectively. The 70 neonates delivered by cesarean section for distress, compared with the 420 without, had a significantly higher incidence of pH < 7.00 (8.5% vs 0.7%, p = 0.0004, relative risk 5.0, 95% confidence interval 2.9 to 8.4). Sensitivity and positive predictive values of an amniotic fluid index < 5th percentile for gestational age to predict pH < 7.00 were 0.8% and 22%, respectively, and for an amniotic fluid index < or = 5.0 cm, 0.5% and 11%, respectively. Receiver-operator characteristic curves indicate that an amniotic fluid index between 0 and 20 cm cannot predict accurately which parturients will have cesarean sections for distress or be delivered of a newborn with a low Apgar score at 5 minutes or a pH < 7.10. CONCLUSION: Both criteria for oligohydramnios are poor predictors of adverse outcome for high-risk intrapartum patients. PMID- 9215166 TI - Cerebrovascular disorders complicating pregnancy--beyond eclampsia. AB - OBJECTIVE: Our purpose was to investigate the problems encountered in the diagnosis and management of cerebrovascular disorders associated with pregnancy and the puerperium. STUDY DESIGN: Pregnancies complicated by cerebrovascular disorders were identified by retrospective chart review (1985 to 1995). Events associated with trauma, neoplasm, drug ingestion, and infection were excluded. RESULTS: The study population comprised 24 women with a variety of cerebrovascular disorders: 14 with infarction (5 arterial, 9 venous), 6 with intracranial hemorrhage (3 anatomic malformation, 3 unknown etiology), 3 with hypertensive encephalopathy, and 1 with an unruptured aneurysm. Blood pressure reflected physical condition at presentation and did not predict diagnosis or outcome except in the 3 women with hypertensive encephalopathy. Only 4 of 14 women with infarction and 1 of 6 with intracranial hemorrhage had a diastolic blood pressure > or = 110 mm Hg. Presumption of eclampsia delayed the diagnosis in 10 women (41.7%). In addition, patient delay in seeking medical attention complicated 10 cases. After review, none of the adverse maternal outcomes were deemed preventable by earlier physician intervention. Seven maternal deaths occurred (29.2%). Neonatal outcome was related to the gestational age and the maternal condition at presentation. CONCLUSION: Cerebrovascular disorders are an uncommon and unpredictable complication of pregnancy that are associated with substantial maternal and fetal mortality. Suspected eclampsia unresponsive to magnesium sulfate therapy warrants an immediate neuroimaging study. Interestingly, in women with intracranial hemorrhage, severe hypertension was not an associated predictive factor. PMID- 9215167 TI - Nucleated red blood cells in cord blood of singleton term neonates. AB - OBJECTIVE: This study aims to establish normal values for nucleated red blood cells in term singletons and factors associated with their elevation. STUDY DESIGN: Cord blood was prospectively collected from term singleton gestations from Feb. 1 to July 31, 1995. Umbilical vein white blood cells and nucleated red blood cells were counted and umbilical arterial pH was determined. Medical records provided maternal and neonatal information. RESULTS: Cord blood from 1112 cases was obtained and evaluated for nucleated red blood cells per 100 white blood cells. Nine outliers were censored (nucleated red blood cells per 100 white blood cells = 126 to 830); five cases were excluded because of missing data. The mean value of nucleated red blood cells per 100 white blood cells was 8.55, the SD was 10.27, and the range was 0 to 89. The value did not very by maternal tobacco or drug use, anemia, fetal presentation, or mode of delivery. Both maternal diabetes and meconium were associated with elevated values, p < 0.01. Apgar scores and cord pHs showed trends toward inverse proportionality to the number of nucleated red blood cells per 100 white blood cells. CONCLUSION: The mean number of nucleated red blood cells per 100 white blood cells was 8.55, with a wide range and SD. Elevated values may be associated with markers of intrauterine hypoxia such as meconium, lower Apgar scores, and lower pH values. PMID- 9215169 TI - Lack of effect of antenatal indomethacin on fetal cerebral blood flow. AB - OBJECTIVE: Our purpose was to investigate fetal cerebral blood flow and the incidence of intraventricular hemorrhage in patients undergoing tocolysis with either indomethacin or magnesium sulfate at < 30 weeks' gestation. STUDY DESIGN: Consenting patients at < 30 weeks' gestation with preterm labor were randomized to receive indomethacin or magnesium sulfate tocolysis. Magnesium sulfate was administered intravenously with an 8 gm loading dose given over the first hour, 4 gm over the second hour, and then a maintenance infusion of 2.5 gm per hour. The infusion was continued for approximately 12 hours after the cessation of uterine contractions. Patients randomized to receive indomethacin were given an initial dose of 50 to 100 mg orally or per rectum, followed by 25 to 50 mg orally every 4 to 6 hours for 24 to 48 hours. Oral tocolytic agents were not used after successful tocolysis. Betamethasone was administered to all patients. Patients underwent fetal cerebral Doppler studies during tocolytic therapy and at least 24 hours after completion of the treatment. RESULTS: Twelve patients were randomized to receive indomethacin and twelve patients were randomized to receive magnesium sulfate. Twenty-one fetuses underwent cerebral Doppler studies in triplicate during and after therapy. The mean gestational age at tocolysis was 27.5 +/- 1.9 weeks for the indomethacin group and 26.4 +/- 1.6 weeks for the magnesium sulfate group (p = 0.14). The middle cerebral artery resistance index for fetuses during indomethacin treatment was 0.73 +/- 0.09, whereas the resistance index after therapy was 0.75 +/- 0.05 (p = 0.49). The resistance index during magnesium sulfate tocolysis was 0.79 +/- 0.04 and after therapy it was 0.76 +/- 0.04 (p = 0.18). There was no significant difference in the resistance index between the groups on or off therapy. In addition, the incidence of intraventricular hemorrhage was similar in both groups. CONCLUSION: These results suggest that indomethacin does not significantly affect fetal cerebral blood flow. If antenatal indomethacin in the preterm fetus increases the risk of intraventricular hemorrhage, it would appear to be by another mechanism. PMID- 9215168 TI - Paraaortic lymph node biopsy: a twenty-year study. AB - OBJECTIVES: Paraaortic lymph node biopsy is a controversial but proved technique to determine the extent of spread of cancers from the uterine cervix or endometrium. This article explores the following questions. Does the presence of positive paraaortic lymph nodes result in modification of the patient's therapy? Does the evidence gained from a paraaortic lymph note biopsy improve patient survival? STUDY DESIGN: Five hundred sixty-eight patients had paraaortic lymph node sampling in conjunction with another operative procedure between 1976 and 1995. Five hundred seven (89.3%) of these patients had either endometrial or cervical cancer. RESULTS: Paraaortic lymph node biopsies led to a survival rate of 9.1% for cervical carcinoma and 46.6% for endometrial carcinoma and were associated with acceptable morbidity. CONCLUSIONS: We believe that paraaortic lymph node biopsies should be part of the routine evaluation of patients with gynecologic cancers. The knowledge gained by this procedure along with appropriately administered radiation therapy can save lives. PMID- 9215170 TI - Evaluating sex chromosome content of sorted human sperm samples with use of dual color fluorescence in situ hybridization. AB - OBJECTIVE: Although most methods for selecting the sex of offspring by sorting spermatozoa are ineffective at shifting the ratio of Y- to X-containing cells, some commercial sources continue to offer such services. Our objective was to evaluate commercially "sorted" samples with use of dual-color fluorescence in situ hybridization and to identify variations in assessment by comparing motile and total sperm populations, donors, observers, and fluorescence in situ hybridization probes. STUDY DESIGN: Cryopreserved sperm from seven anonymous donors were processed as for insemination. Sperm cells from each total sample or motile subfraction were prepared for fluorescence in situ hybridization by incubation with disulfide-reducing agents to expand sperm nuclei. Two sets of X and Y chromosome-specific, fluorophore-labeled deoxyribonucleic acid probes were used. At least 400 nuclei from each preparation were classified independently by three blinded observers. Hybridization efficiency, aneuploidy, and sex chromosome content were evaluated in subsets of five unsorted, five female-oriented, and five male-oriented samples. Total and motile subfractions were compared with eight samples. Fluorescence in situ hybridization probes were compared in five paired unsorted samples. RESULTS: No differences were detected between washed samples and paired motile subfractions. No differences in hybridization and aneuploidy were detected between groups of sorted samples. The Y/X ratio was significantly different between the sorted groups. However, male-oriented samples had a lower Y/X ratio than female-oriented samples did. Observer and probe choice accounted for small but significant variations that did not alter conclusions about the X/Y ratio for sorted samples. CONCLUSION: In a series of 10 sorted samples from one commercial source, dual-color fluorescence in situ hybridization demonstrated a small but significant shift in the sex chromosome ratios among samples. However, this shift was opposite to that expected by the orientation of the sorted samples. PMID- 9215171 TI - Are obstetric interventions such as cervical ripening, induction of labor, amnioinfusion, or amniotomy associated with umbilical cord prolapse? AB - OBJECTIVE: Our purpose was to determine whether intrapartum obstetric interventions are associated with umbilical cord prolapse. STUDY DESIGN: A computer search identified patients who had intrapartum umbilical cord prolapse. Thirty-seven cases were identified between 1990 and 1994 (incidence of 1.85 per 1000). These women were randomly matched to control patients with intact membranes. RESULTS: Patients with umbilical cord prolapse were delivered earlier (34.8 vs 37.1 weeks, p = 0.05). Otherwise, there were no differences between groups regarding the use of cervical ripening, incidence of labor induction, or the use of amnioinfusion and amniotomy. Although cervical dilatation and station were similar between groups at the time of admission, women with umbilical cord prolapse did not have as much descent of the presenting part associated with cervical dilatation and progressive labor compared with control patients. CONCLUSION: By themselves, obstetric interventions of cervical ripening, labor induction, amnioinfusion, and amniotomy do not increase the likelihood that a patient will have umbilical cord prolapse. PMID- 9215172 TI - Subtotal hysterectomy in modern gynecology: a decision analysis. AB - OBJECTIVE: Our purpose was to compare the risks and benefits of subtotal (supracervical) hysterectomy with those of total hysterectomy in women at low risk for cervical cancer. STUDY DESIGN: A decision analysis was performed. Baseline probabilities for operative and postoperative morbidity, mortality, and long-term quality of life were established for subtotal and total hysterectomy. RESULTS: Operative complication rates and ranges for total abdominal hysterectomy were infection 3.0% (3.0% to 20.0%), hemorrhage 2.0% (2.0% to 15.4%), and adjacent organ injury 1.0% (0.7% to 2.0%). Those for subtotal hysterectomy were infection 1.4% (1.0% to 5.0%), hemorrhage 2.0% (0.7% to 4.0%), and adjacent organ injury 0.7% (0.6% to 1.0%). Operative mortality, the risk for development of cervicovaginal cancer, and long-term adverse effects on sexual or vesicourethral function were low in both groups. CONCLUSIONS: Recently proposed benefits from subtotal hysterectomy are not well proven. Total hysterectomy remains the procedure of choice for most women. PMID- 9215173 TI - Do twins mature earlier than singletons? Results from a matched cohort study. AB - OBJECTIVE: Our purpose was to determine whether, as a consequence of advanced maturity, preterm twin infants have a more favorable neonatal outcome than matched singleton infants. STUDY DESIGN: A matched cohort study design was used. Two hundred twenty-four twin infants (112 sets) were matched for gestational age, race, gender, and mode of delivery with singleton infants who were delivered because of refractory preterm labor. Pregnancies with maternal medical disease including preeclampsia, premature rupture of membranes, twin-twin transfusion syndrome, and known fetal anomalies were excluded. Information was obtained by review of maternal and neonatal charts. RESULTS: There was no difference in the incidence of neonatal death (5% vs 7%, p = 0.66), respiratory distress syndrome (38% vs 35%, p = 0.54), grades 3 and 4 intraventricular hemorrhage (5% vs 4%, p = 0.63), grades 2 and 3 necrotizing enterocolitis (4% vs 6%, p = 0.52), and 5 minute Apgar score < or = 6 (21% vs 21%, p = 1.00). Twins had a higher incidence of admission to the Special Care Unit (88% vs 72%, p < 0.001). Results were similar when analysis was limited to presenting twins, nonpresenting twins, and twins concordant with controls for antenatal glucocorticoid exposure. CONCLUSION: Twin infants do not have accelerated maturation and improved neonatal outcome compared with matched singleton infants born at the same gestational age because of preterm labor. PMID- 9215174 TI - The functional anatomy of the urethra: role of the pubourethral ligaments. AB - OBJECTIVE: This clinical study examines and defines the functional anatomy of the urethra as it relates to the Valsalva and Kegel maneuvers and to urethral stability. STUDY DESIGN: Dissection of embalmed cadavers and examination of 60 patients were performed to study adjunct structures in urethral stability. Provocative maneuvers (Valsalva and Kegel) were used in all 60 patients. Urethral prolapse was graded with use of the international Continence-Society classification. RESULTS: Cadaveric dissection confirmed the structural anatomy of the pubourethral muscles and ligaments. Physical examination in 30 patients revealed a lack of urethral stability in all patients with stress urinary incontinence. In 30 patients acting as normal controls, no urinary incontinence was present, and all maintained urethral stability with provocation. The urethrovesical junction was mobile in all patients in performing a Valsalva maneuver. CONCLUSION: Intact pubourethral ligamentous and muscular attachments aid in stabilizing the urethra to its normal anatomic position. This helps maintain continence. PMID- 9215175 TI - Trauma in pregnancy: normal Revised Trauma Score in relation to other markers of maternofetal status--a preliminary study. AB - OBJECTIVE: Our goal was to examine whether a correlation exists between the Revised Trauma Score assigned on admission and pregnancy outcome, as well as whether the Revised Trauma Score has any predictive value for optimal duration of cardiotocographic monitoring necessary to detect immediate adverse pregnancy outcome. STUDY DESIGN: A retrospective chart review was performed of 30 pregnant trauma patients admitted during a 1-year period. Evaluation of cardiotocographic data for either contractions or decelerations or both was performed without knowledge of Revised Trauma Score or maternofetal outcome at discharge. RESULTS: Review of uterine activity and fetal decelerations did not detect useful predictive patterns unless the tracing was immediately ominous, although uterine activity did initially decrease over time. CONCLUSIONS: The Revised Trauma Score lacks predictive value for both risk of adverse pregnancy outcome and need for prolonged cardiotocographic monitoring. A larger patient population needs to be studied for an accurate determination of whether the Revised Trauma Score has potential as a predictive tool. PMID- 9215176 TI - Uterine myomas and factors associated with hysterectomy in premenopausal women. AB - OBJECTIVE: Our purpose was to describe clinical characteristics in premenopausal women with uterine myomas and to identify factors associated with hysterectomy. STUDY DESIGN: Data were collected by chart abstraction in 421 premenopausal women with myomas and analyzed by univariate and multivariable regression. RESULTS: Over a median follow-up period of 29 months, 86% of women had symptoms associated with myomas and 40% had an increase in uterine size of > 2 gestational weeks. By multivariable regression, bleeding symptoms at presentation and previous surgical history of cholecystectomy and adhesiolysis were significantly associated with greater odds of hysterectomy. There was a significant interaction between age and uterine size, so that as age increased, uterine size had a greater impact on the likelihood of hysterectomy. CONCLUSIONS: In this cohort of premenopausal women myomas were associated with symptoms in almost all women over the follow-up period. Hysterectomy was performed in 22% of women overall. PMID- 9215178 TI - Total colpocleisis for vaginal eversion. AB - OBJECTIVE: This report describes a technique for total colpocleisis performed on women with posthysterectomy vaginal eversion and presents the outcome of this surgery. STUDY DESIGN: Thirty-three women, aged 51 to 94 years (78.1 +/- 8.8, average +/- SD) with vaginal eversion were treated with total colpocleisis. Twenty-four women had previously undergone a total of 40 operations for prolapse, and many had a massive prolapse with scarring and ulceration. Five women had stress incontinence, and an additional 12 had poor urethral support without stress incontinence. In association with colpocleisis, 14 had suburethral plication of the endopelvic fascia; two, needle suspensions; and one, a pubovaginal sling. Three had a perineorrhaphy. RESULTS: Operations lasted from 30 to 205 minutes (101 +/- 33.4, average +/- SD), and the estimated blood loss ranged from 20 to 750 ml (206 +/- 171, average +/- SD). No operative complications occurred. Postoperatively, congestive heart failure developed in two women and one had pneumonia; all illnesses resolved with appropriate therapy. There were no complications at the operative site. Average follow-up was 35 (+/ 48) months. All women were initially cured of the vaginal eversion. Recurrent eversion developed in one woman 1 year after surgery and was successfully treated with repeat colpocleisis. Of five women with preoperative stress incontinence, four were cured and one lost to follow-up. No new stress incontinence occurred. CONCLUSION: Total colpocleisis is an effective operation for the treatment of vaginal eversion in selected situations. When defective urethral support is corrected at the time of the operation, postoperative incontinence is not usually a problem. PMID- 9215177 TI - Inadequate weight gain among pregnant adolescents: risk factors and relationship to infant birth weight. AB - OBJECTIVE: Our purpose was to identify behavioral markers for inadequate weight gain (< 20 pounds) during pregnancy among adolescents < 18 years old. STUDY DESIGN: A total of 337 adolescents who were delivered of a term infant at our institution between March 10, 1992, and November 28, 1994 participated in this study. A comprehensive structured interview conducted at the first prenatal visit elicited demographic information and behavioral risk factors. Maternal weights, reproductive history, evidence of sexually transmitted disease, and infant birth weight were extracted from medical records. Logistic regression and chi 2 analyses compared characteristics and infant birth weights between those who gained < 20 pounds with those who gained > or = 20 pounds. RESULTS: A total of 11.6% (39/337) of the total sample gained < 20 pounds during the pregnancy. Adolescents who gained < 20 pounds compared with > or = 20 pounds were delivered of significantly lighter (2942 gm vs 3392 gm) infants and were more likely to be delivered of infants weighing < 2500 gm (13% vs < 1%). Stepwise logistic regression revealed that adolescents who were battered (odds ratio 5.3) or had a sexually transmitted disease (odds ratio 2.3) or an unplanned pregnancy (odds ratio 8.1) were at increased risk for insufficient weight gain during pregnancy. CONCLUSION: Our data suggest that behavioral risk factors are important in the identification of adolescents at greatest risk for inadequate weight gain. Early identification during pregnancy is essential to modify nutritional practices and thus minimize poor obstetric outcomes. PMID- 9215179 TI - Does advanced maternal age affect pregnancy outcome in women with mild hypertension remote from term? AB - OBJECTIVES: Our purpose was to compare maternal and perinatal outcomes of mature women with those in younger women with pregnancies complicated by mild hypertension remote from term. STUDY DESIGN: A matched cohort design was used. A total of 379 mature pregnant women (> or = 35 years old) with mild hypertension remote from term were matched for race, gestational age, and proteinuria status at enrollment with 379 adult controls aged 20 to 30 years also with mild hypertension remote from term. All were enrolled in an outpatient management program that included automated blood pressure measurements and daily assessment of weight, proteinuria, and fetal movement. RESULTS: The mean gestational age at enrollment was 32.7 +/- 3.0 weeks for both groups (range 24 to 36 weeks). By matching 20.6% of patients in each group had > or = 1+ proteinuria on urinary dipstick at enrollment, and 77.3% of patients in each group were white. Chronic hypertension was more common in the mature group (22.4% vs 14.5%, p = 0.007). The mean gestational age at delivery (37.2 +/- 2.3 vs 37.2 +/- 2.2 weeks), the mean pregnancy prolongation (28.1 +/- 21.0 vs 28.4 +/- 22.0 days), and the mean birth weights (2864 +/- 770 vs 2906 +/- 788 gm) were similar between the mature and younger groups (all p > 0.05). There were no differences regarding abruptio placentae (2 vs 3 cases) or thrombocytopenia or HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome (7 vs 9 cases), and there were no cases of eclampsia. There were five stillbirths in the mature group and none in the younger group (p = 0.063). CONCLUSION: Outpatient management of mild hypertension remote from term in the mature pregnant women was associated with similar maternal outcomes but with a nonstatistically higher stillbirth rate compared with the younger pregnant woman. PMID- 9215180 TI - My fifty-year odyssey in obstetrics and gynecology. PMID- 9215181 TI - The effect of placental removal method on the incidence of postcesarean infections. AB - OBJECTIVE: Our purpose was to determine whether the incidence of postoperative endometritis and wound infection is associated with the method of placental removal at the time of cesarean section. STUDY DESIGN: Parturients undergoing cesarean delivery were prospectively randomized to have the placenta removed manually or spontaneously. Patients were excluded from participation if they had received intrapartum prophylactic antibiotics or had been determined to have chorioamnionitis. After delivery of the infant women in the manual group had the placenta extracted by the primary surgeon, whereas women in the spontaneous group had the placenta delivered by gentle traction on the umbilical cord. All study subjects received perioperative prophylactic antibiotics. The primary outcome variable was a postcesarean infection, defined as postecsarean endometritis or wound cellulitis requiring drainage and antibiotic therapy. RESULTS: A total of 333 women were enrolled in the investigation, with 165 assigned to the manual removal group and 168 allocated to have spontaneous removal. There were no statistically significant differences in mean gestational age, frequency or duration of ruptured membranes, frequency or duration of labor, or mean number of vaginal examinations between the two study groups. Postoperative infections occurred in 25 of 168 (15%) women in the spontaneous delivery group compared with 44 of 165 (27%) women in which the placenta was manually extracted (relative risk 0.6, 95% confidence interval 0.4 to 0.9, p = 0.01). Subset analysis of patients delivered with ruptured membranes similarly demonstrated a statistically significant reduction in the incidence of postoperative infections with spontaneous placental removal compared with manual extraction (20% vs. 38%, relative risk 0.5, 95% confidence interval 0.3 to 0.9, p = 0.02). There was a similar trend toward a reduction in postdelivery infections associated with spontaneous placental removal in women with intact membranes; however, this difference did not attain statistical significance. CONCLUSIONS: Spontaneous delivery of the placenta after cesarean delivery is associated with a decrease in the incidence of postcesarean infections. PMID- 9215182 TI - Alteration of ascending thoracic aorta compliance after treatment with menotropin. AB - OBJECTIVE: Our purpose was to determine whether aortic size and compliance are altered by an exogenously induced rise in estrogen. STUDY DESIGN: Magnetic resonance imaging was used to determine the aortic cross-sectional area/aortic pressure relationship in nine premenopausal women before and after menotropin therapy. Simultaneous electrocardiograms, carotid pulse tracings, phonocardiograms, and brachial artery pressures were obtained before each magnetic resonance imaging acquisition. Ascending thoracic aorta cross-sectional area was obtained every 32 msec and aligned with brachial artery pressures extrapolated from the carotid pulse tracing, allowing construction of the ascending thoracic aorta cross-sectional area/aortic pressure relationships. Aortic cross-sectional area was normalized to body surface area, and the shifts in the position for the ascending thoracic aorta cross-sectional area/aortic pressure relationship were determined with use of analysis of covariance. RESULTS: Heart rate and aortic pressure were unchanged before and after menotropin treatment. Initial estradiol levels were < 20 pg/ml. After menotropin treatment (7.4 +/- 1.0 days) estradiol levels rose to 905 +/- 371 pg/ml (p < 0.0001). Ascending thoracic aorta cross-sectional area/body surface area was not significantly increased, adjusted y mean of 389 +/- 7 mm2/m2 before and 403 +/- 7 mm2/m2 after menotropin treatment (p < 0.24). The slope of the ascending aorta cross-sectional area/aortic pressure relationship, an index of aortic compliance, increased from 1.4 +/- 0.6 mm2/m2/mm Hg before to 1.7 +/- 0.6 mm2/m2/mm Hg after menotropin treatment (p < 0.001). CONCLUSION: In premenopausal women a short-term rise in estrogen induced by menotropin treatment is associated with an increase in aortic compliance. Aorta size is not significantly increased within this time frame. PMID- 9215183 TI - The luteal phase of cycles utilizing controlled ovarian hyperstimulation and the possible impact of this hyperstimulation on embryo implantation. AB - OBJECTIVE: Our purpose was to evaluate the early luteal phase of assisted reproductive cycles utilizing controlled ovarian hyperstimulation and to compare these results with those obtained in unstimulated cycles. STUDY DESIGN: We undertook a descriptive study analyzing luteal phase serum progesterone levels, endometrial histologic features, and endometrial surface ultrastructure by scanning electron microscopy of cycles utilizing controlled ovarian hyperstimulation. Study samples were obtained from 7 oocyte donors undergoing controlled ovarian hyperstimulation for the purpose of follicle aspiration in oocyte donation. Control (unstimulated) serum progesterone samples were obtained from 19 patients undergoing in vitro fertilization in unstimulated cycles. Prospective recipients of oocyte donation (n = 20) undergoing mock cycles of exogenous estradiol and progesterone acted as controls for the endometrial biopsies. RESULTS: Serum progesterone levels on the day of human chorionic gonadotropin administration were twofold higher in the study group than in the unstimulated group (1.1 +/- 0.6 vs 0.5 +/- 0.2 ng/ml, mean +/- SD, p < 0.01). On the day of follicle aspiration, progesterone levels were much higher in the study group (8.5 +/- 2.2 vs 0.5 +/- 0.1 ng/ml, p < 0.001). Histologic dating of endometrial biopsies revealed that the study group was advanced by nearly 2 days as compared with the group having artificial cycles. Pinopods, ultrastructural markers of the implantation window, were present in only one of seven study cycles as compared with all of the four artificial cycles. CONCLUSIONS: The early luteal phase of cycles undergoing controlled ovarian hyperstimulation is characterized by markedly elevated serum progesterone levels during the periovulatory period, advanced endometrial histologic features, and an absence of endometrial pinopods at the time of embryo implantation. We speculate that these high levels of progesterone in the early luteal phase cause premature endometrial luteinization and a premature appearance of the implantation window, thus providing an explanation for the observed decrease in endometrial receptivity. PMID- 9215184 TI - Vaginal pH as a marker for bacterial pathogens and menopausal status. AB - OBJECTIVE: Our purpose was to confirm the elevation of vaginal pH expected in patients with bacterial pathogens in premenopausal women and to examine the relationship of serum follicle-stimulating hormone and estradiol levels to vaginal pH in menopausal patients without and with hormone replacement therapy. STUDY DESIGN: Vaginal pH was determined by phenaphthazine (Nitrazine) pH paper in 253 patients seen in a solo private practice for routine speculum examination. None of the patients were pregnant. Measurements were made of serum levels of follicle-stimulating hormone and estradiol for 172 patients and vaginal cultures were taken from 82 patients. Vaginal pH was correlated with vaginal cultures and serum follicle-stimulating hormone and estradiol levels by use of statistical analysis. RESULTS: Vaginal pH was elevated in all premenopausal patients with documented bacterial pathogens. Serum estradiol levels showed an inverse and serum follicle-stimulating hormone levels a direct statistical correlation with vaginal pH in menopausal patients. CONCLUSIONS: Measurement of vaginal pH is useful, effective, and inexpensive for screening purposes. A vaginal pH of 4.5 is consistent with a premenopausal serum estradiol level and the absence of bacterial pathogens. An elevated vaginal pH in the 5.0 to 6.5 range suggests a diagnosis of either bacterial pathogens or decreased serum estradiol. In patients with an elevated pH, vaginal culture should establish the diagnosis. In the absence of bacterial pathogens, a vaginal pH of 6.0 to 7.5 is strongly suggestive of menopause. Titration of estradiol level by vaginal pH during estrogen replacement therapy may help menopausal women avoid side effects or cessation of therapy. PMID- 9215185 TI - Leiomyoma of the female urethra and bladder: report of twenty-three patients and review of the literature. AB - OBJECTIVES: Our purpose was to review what may be the largest experience of bladder and urethral leiomyomas from a single institution. STUDY DESIGN: A retrospective review was done of 23 female patients with emphasis on presentation, symptoms, and operative approach for excision. RESULTS: The majority of bladder and urethral leiomyomas in this series were asymptomatic, nonobstructive, or incidental (discovered at surgery for another entity). Ten patients had a palpable mass on physical examination. Two patients had pain as a presenting complaint. The route of operative excision was transvaginal (10 patients), transurethral (6 patients), or abdominal (6 patients). One patient had the leiomyoma removed elsewhere with a resultant vesicovaginal fistula. CONCLUSIONS: Corollaries should be sought with the experience of uterine leiomyomas, which are histologically identical to bladder leiomyomas. Asymptomatic, nonobstructive, and nonproblematic leiomyomas should not serve as an indication for primary operation. Pedunculated endovesical lesions may be an exception because of the ease of transurethral removal and their tendency to cause future symptoms. Ultrasonographic imaging, cystoscopy, and biopsy should be considered to allow observation and follow-up of leiomyomas. Future investigative cytogenetic studies should be considered on these mesenchymal tumors. PMID- 9215186 TI - Accuracy of mammographic appearances after breast fine-needle aspiration. AB - OBJECTIVE: The objective of this study was to document the observation that fine needle aspiration of palpable breast masses by use of a modified technique performed shortly before mammography need not adversely interfere with mammographic interpretation nor produce falsely suspicious mammographic lesions that delay meaningful evaluation and management in this breast clinic. STUDY DESIGN: In a retrospective record review 1007 women who were seen in the Breast Diagnostic Center at Women's and Children's Hospital from January 1992 until April 1995 and who had fine-needle aspiration of a palpable solid breast mass within 2 weeks before mammography were analyzed overall and in 10-year age group subsets. The mammographic reports of "suspicious" lesions were correlated with having had a prior fine-needle aspiration (within 2 weeks). RESULTS: Of the 1007 women undergoing fine-needle aspirations, 91 had a cytologic or tissue biopsy specimen diagnosis of malignancy. Of these, 72 had "suspicious" mammograms and 19 had "nonsuspicious" mammograms. The calculated positive predictive value was 58%. The negative predictive value was 98%. Mammographic sensitivity was 79%. Specificity was 94%. Age stratification did not reveal any meaningful trends. Of the 916 patients with benign cytologic results of fine-needle aspiration specimens, 52 had "suspicious" mammograms and 864 had "nonsuspicious" mammograms. CONCLUSION: For patient convenience and expeditious diagnosis of a palpable breast mass, fine-needle aspiration can be performed on the initial visit and mammograms subsequently taken within 2 weeks without undue clinical confusion or misleading mammographic findings. Concordance of the diagnostic triad consisting of (1) clinical impression (by history and examination), (2) fine-needle aspiration, and (3) mammography gives a reliable conclusion and can appropriately be used as the basis for clinical management of a breast mass. However, when there is doubt or anxiety about the diagnosis either on the part of the patient or the physician, a definitive histologic tissue diagnosis should obtained. PMID- 9215187 TI - New surgical procedures: can our patients benefit while we learn? AB - Several forces have combined to encourage gynecologic surgeons to acquire the skills they need to perform new endoscopic procedures. Pressures from health care institutions, industry, and, most important, from patients lead to increased demand for less invasive approaches to the treatment of gynecologic conditions. This demand may outstrip the profession's ability to demonstrate the safety and effectiveness of new procedures through rigorous clinical trials. Early on, the benefits expected from laparoscopic surgery may be limited by harms resulting from surgical inexperience. Physicians will struggle to achieve a balance between their ethical obligation to benefit patients while avoiding harm to them and their professional expectation of continued learning. Acquisition of new techniques involves a learning curve, across which complications and operating time decrease while the potential for benefit rises. To minimize harm to patients during the surgeon's learning process, peer review should play an expanded role. Surgeons should discuss their own surgical experience and level of skill openly with their patients as part of the process of informed consent. A relationship of trust is vital when one engages patients in a cooperative educational venture. PMID- 9215188 TI - Hyperreactio luteinalis differentiated from severe ovarian hyperstimulation syndrome in a spontaneously conceived pregnancy. AB - The clinical presentation of hyperreactio luteinalis can mimic ovarian hyperstimulation. Historically, though, the former most often leads to unnecessary surgery whereas the latter is treated supportively. We present a case of a 32-year-old woman who was initially seen with markedly enlarged multicystic ovaries, ascites, and pleural effusions in the tenth week of a spontaneously conceived gestation. Despite a noniatrogenic cause, the patient received supportive management, as would be given with ovarian hyperstimulation syndrome. Making the distinction between hyperreactio luteinalis and ovarian hyperstimulation syndrome has important consequences for diagnosis and management. PMID- 9215189 TI - Preinduction cervical ripening: a randomized prospective comparison of the efficacy and safety of intravaginal and intracervical prostaglandin E2 gel. AB - OBJECTIVE: The purpose of this study was to compare the effectiveness and safety of vaginally administered 5 mg prostaglandin E2 gel prepared in our hospital pharmacy with a commercially available 0.5 mg intracervical prostaglandin E2 gel. STUDY DESIGN: Eighty-three patients undergoing labor induction were randomly assigned to one of two groups for cervical ripening. Either a preparation of 5 mg of prostaglandin E2 was placed vaginally or a commercially available 0.5 mg of prostaglandin E2 was placed intracervically. A maximum of three doses at 6-hour intervals was administered before oxytocin was begun. RESULTS: Among the 83 patients evaluated, 44 were given vaginal gel and 39 were given intracervical gel. No statistically significant differences were observed between the two treatment groups with respect to the incidence of spontaneous versus induced labor, need for oxytocin augmentation, gel-to-induction interval, Bishop score change, maximum oxytocin dose, maximum dilatation rate, length of labor, cesarean section rate, fetal Apgar scores, fetal umbilical vein pH, or fetal umbilical artery pH. CONCLUSIONS: The two prostaglandin E2 formulations appear equivalent in efficacy and safety. Constraints placed on intracervical prostaglandin E2 gel make the vaginal preparation a desirable choice. PMID- 9215190 TI - Clinical utility of saline solution infusion sonohysterography in a primary care obstetric-gynecologic practice. AB - OBJECTIVE: The purpose of this study was to evaluate the role of saline solution infusion sonohysterography in clinical practice in patients with abnormal uterine bleeding. STUDY DESIGN: A prospective case-controlled study was conducted comparing two-dimensional transvaginal imaging with saline solution infusion sonohysterography, endometrial biopsies, and histologic evaluation after surgical procedures. One hundred twenty-four patients with abnormal uterine bleeding were scanned transabdominally and transvaginally. Patients with an increased endometrial thickness or a poorly defined endometrium underwent saline solution infusion sonohysterography. Sterile saline solution, 4 to 10 ml, was injected into the endometrial cavity under direct ultrasonographic visualization. Once the endometrium was expanded, the presence of polyps or myomas and the anterior and posterior endometrial thickness was assessed. RESULTS: Fifty-six patients were noted to have uterine leiomyomas. Eighteen patients had endometrial polyps. Five patients had simple endometrial hyperplasia. Two patients had atypical hyperplasia. No patients in this study had endometrial cancer. No complications occurred in this group of patients. CONCLUSION: Saline solution infusion sonohysterography is a procedure that aids the clinician in the management of abnormal uterine bleeding and may be more cost effective than traditional methods of evaluation. PMID- 9215191 TI - Flow cytometric analysis of lymph node metastases in advanced ovarian cancer: clinical and biologic significance. AB - OBJECTIVE: This study was undertaken to evaluate the deoxyribonucleic acid content and S-phase fraction in advanced epithelial ovarian carcinomas to determine whether lymph node metastases are biologically distinct from peritoneal sites of metastases. STUDY DESIGN: Thirty-five patients with stage III or IV epithelial ovarian cancer who had undergone complete pelvic and paraaortic lymphadenectomy had representative samples from the primary ovarian tumor, peritoneal metastases, and lymph node metastases analyzed by flow cytometry for deoxyribonucleic acid nuclear content and S-phase fraction. RESULTS: Diploid cell lines are found in metastatic lymph nodes (52%) significantly more frequently than in peritoneal metastases (25%, p < 0.02) or in primary ovarian tumors (26%, p < 0.001). The ploidy category frequency distribution of peritoneal metastases mirrors that found in the primary tumor, and both are significantly different from the ploidy category frequency distribution found in metastatic lymph nodes. Heterogeneity among sites is common, being identified in 54% of patients. Peritoneal metastases are more likely to be concordant with the primary tumor (69%) than are lymph node metastases (39%, p < 0.001). Mean S-phase fraction did not differ overall by site but was significantly different between diploid and aneuploid samples by site. Diploid lymph node metastases were found to have the lowest mean S-phase fraction (7.2% +/- 3.3%), and aneuploid lymph node metastases had the highest mean S-phase fraction (22.3% +/- 10.2%). Diploidy of the primary tumor is a positive predictor of long-term survival. Tumoral heterogeneity and lymph node metastases are not related to survival in this group of patients who underwent therapeutic pelvic and aortic lymphadenectomy. CONCLUSIONS: A high proportion of tumor deposits found in metastatic lymph nodes are diploid with a low S-phase fraction. Therapeutic pelvic and aortic lymph node dissection removes disease that, on the basis of flow cytometric characteristics, may be predicted to be resistant to chemotherapy and radiation therapy. PMID- 9215192 TI - Spontaneous liver hematoma in pregnancy not clearly associated with preeclampsia: a case presentation and literature review. AB - Spontaneous liver hemorrhage with formation of subcapsular hematomas and rupture of Glissan's capsule is a rare but often lethal complication of pregnancy. This entity has usually been associated with severe preeclampsia or the HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome. A case of spontaneous subcapsular hematoma of the liver occurring in the third trimester is presented in which the patient probably had neither preeclampsia nor the HELLP syndrome. The literature on liver hematomas in pregnancy published since 1982 when the term HELLP syndrome was coined is reviewed with a focus on the association of liver hematomas with preeclampsia and the HELLP syndrome. Therapy and maternal and neonatal outcomes for this entity are reassessed. PMID- 9215193 TI - Transvaginal ultrasonographic measurement of endometrial thickness in postmenopausal women receiving estrogen replacement therapy. AB - OBJECTIVE: This study was undertaken to evaluate endometrial thickness by transvaginal ultrasonography in asymptomatic postmenopausal women receiving estrogen replacement therapy. The endometrial thickness in this study group was compared with endometrial thickness measurements in a group of women who had abnormal postmenopausal bleeding. The recent literature was reviewed. STUDY DESIGN: Asymptomatic postmenopausal women receiving estrogen replacement, seen for routine examination during the 1-year period from Jan. 1, 1994, to Dec. 31, 1995, had the endometrium evaluated by transvaginal ultrasonography. Women with abnormal postmenopausal bleeding were likewise evaluated and their measurements compared with those of the study group. RESULTS: Twenty-seven different estrogen and estrogen-progestin combinations in 327 asymptomatic women were studied. Additionally, 24 women who were bleeding, not receiving estrogen, and 46 women with abnormal bleeding on estrogen therapy underwent ultrasonography of the endometrium. Endometrial thickness ranged from 1 to 15 mm in women on a regimen of combined estrogen-progestin therapy, 1 to 14 mm in women using sequential estrogen-progestin, and 3 to 15 mm for women receiving unopposed estrogen in the study group. For women with abnormal bleeding not using estrogen, the endometrium measured an average of 12.3 mm (range 2 to 29 mm), with unopposed estrogen 8.3 mm (range 4 to 13 mm), and for estrogen with progestin 6.5 mm (range 2 to 15 mm). Significant pathologic features were found in those women who had bleeding and endometrial measurements between 5.0 and 29 mm. CONCLUSION: There was no significant difference between endometrial thickness measurements in women receiving various combinations of estrogen replacement. In general, expected endometrial measurements can range from 1 to 15 mm. In women with postmenopausal bleeding, however, significant pathologic features may exist with an endometrium measuring as little as 5 mm. PMID- 9215194 TI - Evaluation of sexual misconduct complaints: the Oregon Board of Medical Examiners, 1991 to 1995. AB - OBJECTIVE: In 1991 the Oregon Board of Medical Examiners initiated a separate category for the complaint of sexual misconduct. Investigated complaints of sexual misconduct brought to the Oregon Board of Medical Examiners were analyzed for the years 1991 to 1995 to serve as a baseline. Comparison was made to the Federation of State Medical Boards sexual misconduct data for 1991 and 1992. STUDY DESIGN: One hundred complaints brought against 80 licensees were evaluated by practitioner's degree, age group, sex, specialty, and disposition of complaints for the years 1991 to 1995. The allegations were classified into behavior categories of sexual impropriety, sexual transgression, and sexual violations. RESULTS: Sexual misconduct was the allegation in 5.9% of the complaints investigated for the study period. Oregon had more sexual misconduct complaints than the average reported to the Federation of State Medical Boards for the years 1991 and 1992. Most (72%) complaints came from the patients or their families. Two female physicians (2.4%) had sexual boundary complaints. Sexual misconduct complaints increased by a risk ratio of 1.44 with advancing age by decades. Allegations classified into behavior categories according to severity revealed 39% sexual impropriety, 31% sexual transgression, and 30% sexual violation. Reportable disciplinary actions occurred only with multiple allegations of sexual impropriety (6.5%) and for sexual transgression (27%) whereas sexual violation allegations often had one complainant but there were 54% reportable disciplinary actions. Family practice, obstetrics and gynecology, and psychiatry had the highest incidence of sexual misconduct complaints whereas psychiatry and obstetrics and gynecology had the highest incidence of reportable disciplinary actions. Twenty-five percent of the closed cases resulted in reportable disciplinary actions. This analysis is discussed in relationship to legal and ethical issues and the goal of zero tolerance. CONCLUSIONS: Oregon has a higher percentage of sexual misconduct complaints than the average for 42 states reporting to the Federation of State Medical Boards for the years 1991 and 1992. Analysis of the Oregon Board's experience for the study years will provide a baseline for future evaluation and as an educational resource for the Oregon Board of Medical Examiners and professional and specialty societies. Ethical standards, the reporting and investigative processes, and the legal framework are in place and lessen the incidence of sexual misconduct and work toward zero tolerance. PMID- 9215195 TI - Incidence and outcome of chromosomal mosaicism found at the time of chorionic villus sampling. AB - OBJECTIVE: Chromosomal mosaicism has been reported in about 1% to 3% of chorionic villus sampling specimens. This report provides incidence and outcome information that should be useful in counseling patients found to have mosaicism on chorionic villus sampling. STUDY DESIGN: A retrospective analysis of 11,200 consecutive patients undergoing chorionic villus sampling at the University of California, San Francisco, during the period from Jan. 1, 1984, to June 1, 1996, was undertaken. RESULTS: A total of 140 cases of mosaicism were identified for an incidence of 1.3%. Follow-up information was available for 130 cases, 26 of which (20%) were confirmed in fetal tissue. Confirmation rates for specific types of mosaicism were as follows: autosomal trisomy 7.6%, sex chromosome 25%, structural abnormality 27.3%, and marker chromosome 77.8%. Neonatal outcome was normal in all cases for which pregnancy continued. CONCLUSION: The data indicate that in most cases of chromosomal mosaicism found by chorionic villus sampling the mosaicism is unlikely to be clinically significant in the fetus. PMID- 9215196 TI - Brachial plexus palsy: an old problem revisited again. II. Cases in point. AB - OBJECTIVES: In spite of mounting evidence to the contrary, plaintiffs' expert witnesses continue to maintain that brachial plexus impairment is almost always the result of excessive lateral traction on the head during the last phase of delivery. Case studies are presented to challenge this concept. STUDY DESIGN: Examples encountered in medicolegal consultations were analyzed with this purpose as our focus. RESULTS: Cases of brachial plexus impairment were encountered in which there was no evidence of shoulder dystocia or extreme lateral traction on the fetal head. In one in which shoulder dystocia was recorded, there was also incontrovertible evidence of intrauterine maladaptation. In another, the posterior shoulder was involved. CONCLUSION: To propose that shoulder dystocia with extreme lateral traction on the fetal head after its delivery is not a factor in some cases of brachial plexus impairment would be insupportable. Conversely, to maintain a posteriori that brachial plexus impairment in itself is evidence that such pressure must have been used is untenable. PMID- 9215197 TI - Vaginal sacrospinous ligament fixation with the Autosuture Endostitch device. AB - OBJECTIVE: The purpose of the study was to evaluate a disposable suturing device to facilitate vaginal sacrospinous ligament fixation. STUDY DESIGN: Seventeen consecutive patients (mean age 66.3 years) requiring vaginal sacrospinous ligament fixation had the procedure performed with the Autosuture Endostitch device with a braided polyester suture. Patients were evaluated with respect to operative time, blood loss, complications, hospital stay, and success of the vaginal fixation. RESULTS: All patients underwent additional procedures, including anterior colporrhaphy (82.4%), posterior colporrhaphy (100%), vaginal hysterectomy (5.9%), enterocele repair (76.4%), and Burch suprapubic urethropexy (5.9%). The time required for the sacrospinous ligament pilcation ranged between 14 and 25 minutes (mean 18.8 +/- 3.0 minutes). Fifteen patients (88.2%) had an estimated blood loss < or = 100 ml for the complete procedure. No complications occurred. No patient was hospitalized > 4 days. Patients were followed up between 2 and 18 months (mean 9.8 +/- 4.2 months). Fifteen patients (88.2%) maintained good vaginal vault support. Two patients (11.8%) had recurrence at 4 and 6 months, respectively. CONCLUSION: The Autosuture Endostitch device, although designed for endoscopic surgery, is efficacious for the performance of sacrospinous ligament fixation of the vaginal vault. Decreasing the length of the instrument would make it even more practical. PMID- 9215198 TI - An obstetric and gynecologic clerkship's influence on a medical community. The Washington, Alaska, Montana, and Idaho Anchorage obstetric and gynecologic clerkship. AB - OBJECTIVES: Our purpose was to explore the influences of an obstetric and gynecologic medical student clerkship on a remote medical community. Return of physicians to Alaska and faculty perceptions of their experience were central foci. STUDY DESIGN: Data were obtained on former clerks to determine choice of specialty and location of practice. Data regarding all physicians new to Alaska was correlated with the University of Washington Medical School graduate data. Additionally, a questionnaire with a Likert-type scale evaluated the 10 clinical faculty members participating in the clerkship. RESULTS: Between 1978 and 1991 we trained 266 clerks. A total of 77 of 374 (21%) new physicians in Alaska (1978 to 1991) were graduates of the University of Washington; 26 of those 77 (34%) were our former Anchorage obstetrics and gynecology clerks. The clinical faculty reported both positive and negative effects of their participation in the clerkship. CONCLUSION: The desired benefit, the return of new physicians to Alaska, seemed supported. Questionnaire results hinted at additional benefits for the supervising faculty physicians in this isolated community. The formal affiliation effected by the clerkship seemed to have a positive impact on patient care, communication, consultation, and shared action among the participating physicians. PMID- 9215199 TI - The Women's Health Curriculum by a problem-based learning method for medical students at the University of California, San Francisco. AB - OBJECTIVES: Our objectives were to (1) expand and strengthen the women's health curriculum at the University of California, San Francisco, and (2) evaluate the responses of both medical students and faculty to this curriculum. STUDY DESIGN: A written evaluation of the curriculum in women's health was completed by both students and faculty. Variables studied included mean scores of cases, the overall course score, and the preferences of medical students for faculty specialty in teaching the small groups. RESULTS: The overall course evaluation score was 7.81 (range 1 to 10). For those students who had both faculty from internal medicine or family medicine and obstetrics and gynecology, there was a strong preference that obstetrician-gynecologists teach the majority of the cases. CONCLUSIONS: The new case-based curriculum in women's health was enthusiastically received by both medical students and faculty. PMID- 9215200 TI - Colposacropexy: an alternative technique. AB - Colposacropexy procedures restore anatomically correct apical vaginal support on the levator plate at the ischial spine level. Venous hemorrhage resulting from laceration of presacral veins during suture fixation is the major hazard of this procedure. Titanium orthopedic bone anchor fixation minimizes this risk through precision placement of the bone anchor-suture unit. PMID- 9215201 TI - The impact of contraceptive methods on the onset of symptomatic vulvovaginal candidiasis within the menstrual cycle. AB - OBJECTIVE: Vulvovaginal candidiasis is the second most common cause of vaginal discharge. Low-dose oral contraceptives are no longer thought to increase the absolute risk of episodic vulvovaginal candidiasis. This study investigates the possible impact that hormonal contraception may have on the timing of onset of symptoms within the menstrual cycle. STUDY DESIGN: In a retrospective chart review of reproductive-aged women seen at the Women's Health Care Clinic at Harbor-University of California, Los Angeles, Medical Center, data from the records of 448 symptomatic women who had 507 episodes of vulvovaginal candidiasis were extracted and analyzed for timing of onset of symptoms within the menstrual cycle. Diagnosis was based on symptoms, physical findings, and microscopy. Onset was divided into five physiologic ranges within an idealized 28-day menstrual cycle. Comparisons among groups were made with use of chi 2 and p < 0.05 thresholds for statistical significance. RESULTS: No differences were found in the onset of symptoms within the idealized menstrual cycle ranges between women using hormonal birth control methods and those using nonhormonal ones. The distribution was remarkably uniform throughout the cycle with the exception of the first few days (during menses). CONCLUSION: The timing of onset of symptoms of vulvovaginal candidiasis within a woman's menstrual cycle is not affected by her method of birth control. PMID- 9215202 TI - Choroid plexus cysts--association with trisomy: prospective review of 16,059 patients. AB - OBJECTIVE: The purpose of the study was to determine the incidence of isolated choroid plexus cysts in association with trisomy 18 and other abnormalities. STUDY DESIGN: All patients from June 1992 through December 1995 were followed up after a screening ultrasonography. Any patient with a choroid plexus cyst was offered genetic counseling and an amniocentesis. Screening ultrasonographic examinations were performed on 16,059 patients, and 301 patients had a fetus with a choroid plexus cyst. One hundred thirty patients elected to have an amniocentesis. Patients were followed up to delivery. RESULTS: Two hundred sixty three patients had an isolated choroid plexus cyst. Thirty-eight patients had a choroid plexus cyst associated with additional risk factors. Risk factors included advanced maternal age, additional ultrasonographic abnormalities, past obstetric history, or family history. No abnormalities were noted in the group with an isolated choroid plexus cyst. Four patients had an abnormality when the choroid plexus cyst was associated with an additional risk factor, including two patients with trisomy 18 and one with trisomy 21. CONCLUSION: An isolated choroid plexus cyst was not associated with a trisomy or other abnormalities in this study. When a choroid plexus cyst was associated with an additional risk factor, 10.5% of the patients had an abnormality. Amniocentesis is recommended when a choroid plexus cyst is found in association with additional risk factors. PMID- 9215203 TI - Transvaginal reduction of an interstitial heterotopic pregnancy with preservation of the intrauterine gestation. AB - With use of transvaginal ultrasonographic guidance, cardiac activity in an interstitial heterotopic pregnancy at 7 weeks' gestation was terminated. The interstitial pregnancy resolved, and a healthy term infant was delivered. If an early diagnosis of an interstitial heterotopic pregnancy is made, selective reduction may allow preservation of the intrauterine gestation without surgery. PMID- 9215204 TI - A randomized controlled trial of a new fetal acoustic stimulation test for fetal well-being. AB - OBJECTIVES: Our purpose was to determine (1) whether a fetal acoustic stimulation test results in more palpable fetal movement compared with a mock test (control) and (2) whether palpated fetal movements after a fetal acoustic stimulation test are accompanied by a reactive nonstress test. STUDY DESIGN: In a randomized controlled trial we studied women seen in the labor and delivery suite for various indications. Women were excluded for multiple gestation, < 31 weeks' gestational age, treatment with magnesium sulfate or narcotics, or ruptured membranes. Informed consent was obtained from eligible women, who were then randomized to a test or control group. We placed an acoustic stimulator on the abdomen of each woman, but only the test group was stimulated. We assessed fetal movement by a grading system: 0 = no fetal movement felt by patient or tester, 1 = fetal movement felt by patient only, 2 = fetal movement felt by tester, 3 = visual movement seen by tester. A positive fetal acoustic stimulation test result was defined as one with any fetal movement felt or seen by the tester (grades 2 or 3). We then performed a nonstress test. We compared rates of a positive fetal acoustic stimulation test in the test and control groups with the chi 2 test. A p value < 0.05 was considered significant. RESULTS: We randomized 297 women to the test group and 280 women to the control (mock test) group. Of women tested with the fetal acoustic stimulation test. 81% had fetal movement by palpation or visualization (grades 2 or 3) compared with 19% of the control group (p < 0.0001, odds ratio 19.29, 95% confidence interval 12.42 to 30.07). Of the test group, 283 (95%) had a reactive nonstress test and 14 (5%) had nonreactive tests; the control group had 267 (95%) reactive and 13 (5%) nonreactive nonstress tests. Of 242 patients in the test group with a positive fetal acoustic stimulation test, 236 (98%) had a reactive nonstress test. Of those in the test group with fewer than three contractions per 10 minutes. 164 (89%) had a positive fetal acoustic stimulation test. Of these, 162 (99%) had a reactive nonstress test. CONCLUSION: The fetal acoustic stimulation test evokes significantly more palpated or visualized fetal movement than in controls. Palpated or visualized fetal movement after acoustic stimulation was almost always accompanied by a reactive nonstress test. PMID- 9215205 TI - Surgical intensive care unit care after ovarian cancer surgery: an analysis of indications. AB - OBJECTIVE: Our purpose was to develop a profile of preoperative and perioperative characteristics that would enable gynecologic oncologists to identify those patients with ovarian cancer who would benefit most from postoperative surgical intensive care unit care and thereby optimize resource utilization and cost effectiveness. STUDY DESIGN: A retrospective analysis was performed of 85 patients admitted to the surgical intensive care unit after cytoreductive surgery between Jan. 1, 1989, and Dec. 31, 1993. Fifty-three patients admitted to the surgical intensive care unit for < 24 hours were compared with 32 patients admitted for > 24 hours. Five preoperative characteristics (age, American Society of Anaesthesiology classification, body mass index, albumin, primary versus recurrent disease) and six perioperative characteristics (estimated blood loss, ascites, surgical time, bowel resection, Swan-Ganz catheter, ventilator dependence) were compared across the two groups by univariate analysis and multivariate logistic regression analyses. RESULTS: All preoperative variables were similar across the two groups. Ascites volume and length of surgery were not significant, whereas estimated blood loss was significant in the univariate analysis but not in the logistic regression analysis. Three perioperative variables were found to be predictive of extended surgical intensive care unit care by logistic regression analysis: placement of a Swan-Ganz catheter (odds ratio 4.31, 95% confidence interval 1.13 to 16.4), bowel resection (odds ratio 13.0, 95% confidence interval 1.96 to 86.5), and ventilator dependence (excluded from logistic regression analysis for mathematic reasons). CONCLUSIONS: The patient's preoperative medical condition proved to be less important than how she fares during surgery. The patient most likely to benefit from surgical intensive care unit care had undergone bowel resection, required invasive hemodynamic monitoring, or was ventilator dependent postoperatively. This patient profile may prove to be a useful screening tool to optimize resource utilization and cost effectiveness, but it cannot replace clinical judgment. PMID- 9215206 TI - Modified Gittes's needle colposuspension method complemented with the laparoscopic ventrosuspension of the uterus for the treatment of stress incontinence associated with stage III or IV pelvic organ prolapse. PMID- 9215207 TI - Acupressure for hyperemesis gravidarum. PMID- 9215208 TI - Do gravid women with preeclampsia-eclampsia ever require plasmapheresis? PMID- 9215209 TI - The microcontact peritoneoscopy: a new diagnostic tool in endometriosis. PMID- 9215210 TI - The vacuum extraction to forceps in posterior presentation comparison. PMID- 9215211 TI - Diagnostic accuracy of outpatient hysteroscopy. PMID- 9215212 TI - Record of the first physician to see Henrietta Lacks at the Johns Hopkins Hospital: history of the beginning of the HeLa cell line. PMID- 9215214 TI - Racial differences in breast cancer survival: the interaction of socioeconomic status and tumor biology. AB - OBJECTIVE: Our purpose was to evaluate the effect of sociodemographic and clinical variables on survival rates of African-American and white women with breast cancer. STUDY DESIGN: Between 1988 and 1992 the Metropolitan Detroit Cancer Surveillance System Identified 10,502 women (82% white and 18% African American) in whom invasive breast cancer was diagnosed. Cox proportional hazards regression was used to estimate the relative risk of death for African-Americans compared with whites after controlling for variables believed to influence survival. RESULTS: African-American women were more likely than white women to have tumors that were of a more advanced stage, a higher grade, and hormone receptor-negative. After controlling for age, tumor size, stage, histologic grade, census-derived socioeconomic status, and the presence of a residency training program at the treatment hospital, the relative risk of dying for African-Americans compared with whites was 1.68 (95% confidence interval, 1.27 2.23) for women less than 50 years of age, and 1.33 (95% confidence interval, 1.13-1.56) for women older than 50 years of age. CONCLUSIONS: Known factors that predict survival differences between African-Americans and whites are more prevalent among women less than 50 years of age, emphasizing the need to focus more attention on public health efforts directed toward younger women. PMID- 9215213 TI - Racial disparity in overexpression of the p53 tumor suppressor gene in stage I endometrial cancer. AB - OBJECTIVE: This study was conducted to determine whether overexpression of the p53 tumor suppressor gene is associated with poor outcome in early-stage endometrial cancers and whether a racial difference in the frequency of p53 overexpression contributes to the observed racial disparity in survival rates. STUDY DESIGN: Immunostaining for the p53 gene was performed in 164 women with stage I endometrial adenocarcinomas. RESULTS: Overexpression of mutant p53 protein was seen in 28 out of 164 (17%) cases and was associated with a poor histologic grade (p = 0.003) and a nonendometrioid histologic appearance (p = 0.06). Overexpression also was three times more frequent in blacks (15 out of 44, 34%) than in whites (13 out of 117, 11%) (p = 0.003). Recurrent disease developed in 15 out of 164 (9%) cases and was more than twice as frequent in cases when the p53 gene was overexpressed (5 out of 28, 18%) than in cases with normal expression (10 out of 136, 7%). Recurrent disease was seen in 6 out of 44 (14%) blacks compared to 9 out of 117 (8%) whites. CONCLUSIONS: These data support the hypothesis that differences in the frequency of alteration of the p53 tumor suppressor gene contribute to the racial disparity in endometrial cancer survival. PMID- 9215215 TI - Establishment of breast cell cultures and lines from peoples of African origin. AB - OBJECTIVES: The purpose of our study was to establish breast epithelial cell cultures and cell lines from peoples of African origin (blacks). It is presumed that the biology of breast cancer in women of African origin has unique aspects that can be explored using cultured breast epithelial cells. STUDY DESIGN: Biopsy specimens were obtained from black women undergoing radical or modified mastectomies. Normal cell cultures were established using tissue from reduction mammaplasties or the milk of lactating mothers. The tissue specimens were lacerated, digested with collagenase solution, and plated on tissue culture plates. To extend the life of the epithelial cells in culture, they are transformed with SV40 virus. RESULTS: We have maintained breast tumor cells in culture from a 27-year-old black woman for more than 1 month. CONCLUSION: Despite the difficulty of establishing epithelial cell cultures, we have maintained breast tumor cells from blacks in culture for an extended period to allow characterization. PMID- 9215216 TI - Characteristics of antigens from an HCG alpha secreting cell line (ElCo) derived from human breast carcinoma in a Native American patient. AB - OBJECTIVE: Our purpose is to report a cell line derived from a Native American patients that could serve as a model for ethnic diversity in cell culture research. STUDY DESIGN: Tumor biopsy specimens from reproductive tract malignancies were explanted in the cell culture laboratory. Characterization of a successfully established line included a parameter commonly observed in minority populations in rapid moving Type A glucose-6-phosphate dehydrogenase enzyme (G6 PD). Multiple other standard characterization studies were carried out. RESULTS: A breast cancer cell line with a G6-PD-A enzyme was derived from an American Indian patient; this pattern is commonly observed in African-American populations. Results of lymphocyte-mediated cytotoxicity tests indicated that lymphocytes from patients with breast cancer were cytotoxic to ElCo cells. On the other hand, lymphocytes from patients with other types of cancer were not significantly different from healthy donor lymphocytes in their cytotoxicity. These studies indicate that ElCo cells express tumor-associated antigen(s). Many antigens of tumor cell origin were detected with use of antisera raised in rabbits in concentrates of ElCo cell culture fluid. One of these antigens was the alpha subunit of human chorionic gonadotropin. CONCLUSIONS: Chemotherapy testing, vaccine production, and various new treatment schemes are most commonly developed in cell culture systems. These systems should reflect characteristics of the target population so that desired outcomes best fit the population being served. Thus the cell line being reported, which was derived from a Native American woman, represents a tool that can be used in cell culture research as a model for diversity. PMID- 9215217 TI - Dietary factors and cancers of breast, endometrium, and ovary: strategies for modifying fat intake in African American women. AB - Modification of dietary fat and fiber could help prevent cancers of the breast, endometrium, and ovary that are prevalent in African-American women. Dietary intervention programs aimed at reducing fat intake have had mixed results in this population. The transtheoretic model is proposed for achieving dietary change. Strategies for changing health behaviors in African-American women include heightening sensitivity to cultural values among health educators and the use of multiple strategies to reinforce messages. To stimulate healthier eating, it is important to incorporate the distinct habitual eating patterns into innovative intervention methods, using effective behavioral change methods. PMID- 9215218 TI - Body mass and 26-year risk of mortality among women who never smoked: findings from the Adventist Mortality Study. AB - The authors have examined the relation between the Quetelet body mass index (BMI) and 26-year risk of all-cause mortality in a population of 12,576 non-Hispanic while, Seventh-day Adventist women (ages 30-74 years) who never smoked. Mortality risk for each BMI quintile (I, < 21.3 kg/m2; II, 21.3-22.9 kg/m2; III, 23.0-24.8 kg/m2; IV, 24.9-27.4 kg/m2; and V, > 27.4 kg/m2) was determined from a proportional hazard regression with adjustment for age and other covariables. In this population, the overall BMI-mortality relation showed dependence on age, duration of follow-up, and baseline indicators of preexisting illness (weight fluctuation, history of major chronic disease, and severe physical complaints). Therefore, the analysis focused on women with no indicators of preexisting illness, and risk estimates were stratified by age at baseline and duration of follow-up. Among middle-aged women (ages 30-54 years), the authors found a weak linear relation during years 1-8 (median attained age, 51 years), a significant linear relation during years 9-14 (median attained age, 57 years), and a significant nonlinear (U-shaped) relation during years 15-26 (median attained age, 68 years). Among older women (ages 55-74 years), they found a significant nonlinear (U-shaped) relation during years 1-8 (median attained age, 71 years) and significant linear relations during years 9-14 (median attained age, 77 years) and years 15-26 (median attained age, 87 years). These findings implicate overweight as a risk factor for fatal disease among women throughout adulthood and raise the possibility that lean, apparently healthy, middle-aged women may experience a higher risk of death during old age due to their lower body weight. PMID- 9215219 TI - Relative contributions of incidence and survival to increasing prevalence of adult-onset diabetes mellitus: a population-based study. AB - This population-based retrospective study investigates temporal trends in adult onset diabetes mellitus prevalence, incidence, and survival. The complete community-based medical records, including laboratory results, of all Rochester, Minnesota, residents with a clinical diagnosis of diabetes or diabetes-like condition were reviewed to identify incidence cases aged 30 years or more from 1945 to 1989 (n = 1,847) and prevalence cases aged 45 years or more on January 1, 1970 (n = 465), January 1, 1980 (n = 689), or January 1, 1990 (n = 973). Glucose values and case definitions were standardized throughout. Observed 10-year survival for 1970 and 1980 prevalence cases was compared with that expected for Minnesota white populations in 1970 and 1980, respectively. Age-adjusted prevalence rose 65% for men and 37% for women between 1970 and 1990. There were marked differences among prevalence groups in treatment type, the proportion diagnosed using glucose tolerance tests, and the proportion categorized as obese. Relative survival for 1980 prevalence cases was not greater than that for 1970 prevalence cases. Age-adjusted incidence rates rose 47% for men and 26% for women between 1960 and 1965 and 1985 and 1989. These findings emphasize the need for heightened surveillance and intervention to reduce the burden of illness from adult-onset diabetes mellitus in the population. PMID- 9215220 TI - Relation between birth weight and the insulin sensitivity index in a population sample of 331 young, healthy Caucasians. AB - The objective was to study the association between birth weight and the insulin sensitivity index. Altogether, 331 unrelated Caucasian subjects aged 18-32 years with measures of the insulin sensitivity index and insulin secretion during a combined intravenous glucose and tolbutamide tolerance test were included in the study. The data on birth weight and length were obtained from the midwife records. The study took place in Copenhagen, Denmark, during 1992-1993. Univariately, a significant positive association between birth weight and the insulin sensitivity index was found in women (p = 0.045), but not in men (p = 0.23). In multivariate analysis, controlling for age, gender, body mass index, waist circumference, maximal aerobic capacity, and use of oral contraceptives, no significant interaction between birth weight and gender that considered the insulin sensitivity index was found. The insulin sensitivity index was significantly associated with birth weight (p = 0.0012), corresponding to an increase of 1.7% (95% confidence interval 0.7-2.7%) in the insulin sensitivity index for each 100-g increases in birth weight. In comparison, an increase in body mass index of 1 kg/m2 (a weight gain of 2.9 kg in a man 1.70 m tall) corresponds to a decrease in the insulin sensitivity index of 3.8% (95% confidence interval 0.7-6.8%), an increase in waist circumference of 1 cm corresponds to a decrease in the insulin sensitivity index of 2.1% (95% confidence interval 0.9-3.1%), and use of oral contraceptives corresponds to a decrease in the insulin sensitivity index of 26.7% (95% confidence interval 12.2 38.1%). Thus, the impact of birth weight on the insulin sensitivity index was of minor importance. PMID- 9215221 TI - Effects of cigarette smoking, caffeine consumption, and alcohol intake on fecundability. AB - Data from the Ontario Farm Family Health Study were analyzed to determine whether smoking, caffeine, or alcohol use among men and women affect fecundability (the monthly probability of conception). In this retrospective cohort study of farm couples in Ontario, Canada, the farm operator, husband, and wife completed questionnaires during 1991-1992, yielding information on 2,607 planned pregnancies that had occurred over the previous 30 years. Fecundability ratios were calculated using an analog of the Cox proportional hazards model. Cigarette smoking among women and men was associated with decreased fecundability (fecundability ratio = 0.90, 95% confidence interval (CI) 0.82-0.98 and fecundability ratio = 0.88, 95% CI 0.81-0.95, respectively). Caffeine consumption of 100 mg or less versus more than 100 mg in women and men was not associated with fecundability (fecundability ratio = 0.98, 95% CI 0.91-1.07 and fecundability ratio = 1.05, 95% CI 0.97-1.14, respectively). Decreases were observed among women who were coffee drinkers (fecundability ratio = 0.92, 95% CI 0.84-1.00) and men who were heavy tea drinkers (fecundability ratio = 0.85, 95% CI 0.69-1.05), regardless of caffeine content. Alcohol use among women and men was not associated with fecundability. These data are consistent with previous studies of the adverse effect of tobacco on fecundability in female smokers and suggest an effect of smoking among males. Continued evaluation of coffee and tea is warranted to address constituents other than caffeine. PMID- 9215222 TI - Exposure to environmental tobacco smoke and birth outcome: increased effects on pregnant women aged 30 years or older. AB - The purposes of this study were to examine the association between self-reported environmental tobacco smoke (ETS) exposure during pregnancy and birth weight, prematurity, and small-for-gestational age infants and to determine whether these associations differ by maternal age. Data from the Pregnancy Nutrition Surveillance System from two states that collected data on both passive and active smoking for the period 1989-1994 were analyzed. ETS exposure was defined as reported exposure to the cigarette smoke of a household member. Multiple logistic and linear regression analyses were used to evaluate the association between ETS and birth outcomes. The mean adjusted birth weight among infants of nonsmoking mothers age 30 years or older was 90 g less among infants exposed to ETS than among infants not exposed. No significant association was found among infants of younger nonsmoking mothers. Similarly, the risks for low birth weight (adjusted odds ratio (OR) = 2.42, 95% confidence interval 1.51-3.87) and preterm delivery (adjusted OR = 1.88, 95% confidence interval 1.22-2.88) were elevated among older nonsmokers exposed to ETS, but not among younger nonsmokers exposed to ETS (adjusted OR = 0.97, 95% confidence interval 0.76-1.23; adjusted OR = 0.92, 95% confidence interval 0.76-1.13, for low birth weight and preterm delivery, respectively). These findings indicate that the association between ETS exposure and adverse pregnancy outcomes appears to be modified by maternal age. PMID- 9215223 TI - Neighborhood environments and coronary heart disease: a multilevel analysis. AB - The authors investigated whether neighborhood socioeconomic characteristics are associated with coronary heart disease prevalence and risk factors, whether these associations persist after adjustment for individual-level social class indicators, and whether the effects of individual-level indicators vary across neighborhoods. The study sample consisted of 12,601 persons in four US communities (Washington County, Maryland; Forsyth County, North Carolina; Minneapolis, Minnesota; and Jackson, Mississippi) participating in the baseline examination of the Atherosclerosis Risk in Communities Study (1987-1989). Neighborhood characteristics were obtained from 1990 US Census block-group measures. Multilevel models were used to estimate associations with neighborhood variables after adjustment for individual-level indicators of social class. Living in deprived neighborhoods was associated with increased prevalence of coronary heart disease and increased levels of risk factors, with associations generally persisting after adjustment for individual-level variables. Inconsistent associations were documented for serum cholesterol and disease prevalence in African-American men. For Jackson African-American men living in poor neighborhoods, coronary heart disease prevalence decreased as neighborhood characteristics worsened. Additionally, in African-American men from Jackson, low social class was associated with increased serum cholesterol in "richer" neighborhoods but decreased serum cholesterol in "poorer" neighborhoods. Neighborhood environments may be one of the pathways through which social structure shapes coronary heart disease risk. PMID- 9215225 TI - Prevalence of melanocytic nevi and freckles in young Israeli males. Correlation with melanoma incidence in Jewish migrants: demographic and host factors. AB - The role of host and environmental factors in the pathogenesis of multiple melanocytic nevi, atypical nevi, and freckles was studied in 1989 in a random sample of 3,040 Israeli males aged 17 years. Multiple melanocytic nevi were significantly associated with family history of melanoma or multiple melanocytic nevi (odds ratio (OR) = 15.0), fair or lightly pigmented skin color (OR = 2.7 and 2.3, respectively), and affiliation to the high or heterogenous melanoma risk group, determined by the incidence rates of melanoma in Jewish migrants from corresponding origin (OR = 3.1 and 2.1, respectively). An environment-related effect may account for the increased multiple melanocytic nevi risk among second- (OR = 8.2) compared with first-generation, native-born recruits (OR = 3.0) from the high melanoma risk group whose families had been living in Israel the longest. Atypical nevi were associated with fair (OR = 6.1) and lightly pigmented (OR = 3.5) skin color, high and moderate sunburn susceptibility (OR = 4.7 and 2.5, respectively), and family history of melanoma or multiple melanocytic nevi (OR = 4.7). Freckles were significantly associated with sun-sensitive phenotype, family history of melanoma or multiple melanocytic nevi (OR = 1.5). Conservative (OR = 1.9) or nonreligious status (OR = 1.9), and high (OR = 2.4) or heterogenous melanoma risk groups (OR = 1.8). These findings indicate that environmental factors may modify the occurrence of multiple melanocytic nevi and freckles in genetically susceptible ethnic groups. PMID- 9215224 TI - Trends in serum lipid levels during 1980-1992 in children and young adults. The Cardiovascular Risk in Young Finns Study. AB - To assess secular trends in serum lipid levels in Finnish children and young adults, the authors examined a total of 3,517, 2,769, 2,392, 352, and 880 subjects who had complete data on serum lipids in 1980, 1983, 1986, 1989, and 1992, respectively, in a longitudinal follow-up study. Trend analyses were carried out among subjects aged 15 (n = 1,835) or 18 (n = 1,562) years to exclude the confounding effect of age on the study variables. Data on obesity, physical activity, smoking, and alcohol use were available from each study year, and data on diet were available for the study years 1980, 1986, and 1992. Between 1980 and 1992, mean total cholesterol levels decreased from 4.88 to 4.47 mmol/liter (from 189 to 173 mg/dl), and low density lipoprotein cholesterol levels decreased from 3.06 to 2.85 mmol/liter (from 119 to 110 mg/dl). The mean high density lipoprotein cholesterol levels decreased by 19%, from 1.43 to 1.15 mmol/liter (55.2 to 44.6 mg/dl). During 1986-1992, triglyceride levels increased by 15%, from 0.88 to 1.01 mmol/liter (78.2 to 89.9 mg/dl). During 1980-1992, body mass index values increased from 20.8 to 21.8 kg/m2, parallel to increases in skinfold thickness. In the diet, the ratio of polyunsaturated to saturated fatty acids increased from 0.26 to 0.39. Alcohol and oral contraceptive use became more frequent, and the subjects tended to become less physically active. In conclusion, a change in the lipid profile in Finnish adolescents aged 15 and 18 years and young adults during 1980-1992 was observed, characterized by a decrease in low density lipoprotein cholesterol and high density lipoprotein cholesterol levels and an increase in triglyceride level. Possible determinants for these changes include alterations in diet and a trend toward increased obesity. PMID- 9215226 TI - Leukemia risk and occupational electric field exposure in Los Angeles County, California. AB - The authors analyzed data on electric fields from a prior study of occupational magnetic field exposure and leukemia risk conducted in Los Angeles County, California, in 1972-1990. Ranking of exposure differed somewhat for magnetic and electric fields. The odds ratios were 1.22 (95% confidence interval (CI) 0.80 1.86) and 1.15 (95% confidence interval 0.78-1.72) for medium and high exposure categories, respectively, and there was no clear evidence of an exposure-response relation (odds ratio for 10 V/m increase = 1.05, 95% CI 0.95-1.16). Although not conclusive, our analyses provide little support for an association between occupational electric field exposure and leukemia. PMID- 9215227 TI - Household and dwelling contact as risk factors for leprosy in northern Malawi. AB - Data on household and dwelling contact with known leprosy cases were available on more than 80,000 initially disease-free individuals followed up during the 1980s in a rural district of northern Malawi. A total of 331 new cases of leprosy were diagnosed among them. Individuals recorded as living in household or dwelling contact with multibacillary patients at the start of follow-up were at approximately five- to eightfold increased risk of leprosy, respectively, compared with individuals not living in such households or dwellings. Individuals living in household or dwelling contact with paucibacillary cases were both at approximately twofold increased risk. The higher risk associated with multibacillary contact and the fact that dwelling contact entailed a greater risk than household contact if the association was with multibacillary, but not with paucibacillary, disease suggest that paucibacillary cases may not themselves be sources of transmission, but rather just markers that a household has had contact with some (outside) source of infection. When household contact was considered alone, the risks of disease were appreciably higher for younger than for older contacts and for male compared with female contacts. Despite the elevated risk of leprosy associated with household or dwelling contact, only 15% of all incidence cases arose among recognized household contacts. Given the dynamic nature of household membership and consequent misclassification of contact status, the true contribution to overall incidence of contact within household or dwelling settings is likely to be much higher than this, perhaps 30% or higher. Considering the predilection of males for infectious multibacillary forms of the disease, the transmission of Mycobacterium leprae at an early age, in particular to males, may be of particular importance for the persistence of leprosy in endemic communities. Although residential contact with a multibacillary case is the strongest known determinant of leprosy risk, the vast majority of such contacts never manifest disease, which indicates a crucial role for genetic and/or environmental factors in the transmission of M. leprae infection and/or the pathogenesis of clinical leprosy. PMID- 9215228 TI - Bilateral partial anomalous pulmonary venous return. AB - Two unusual cases of atrial septal defects with bilateral partial anomalous pulmonary venous return are described. Total repair was achieved in both children by direct connection of the left upper lobe vein to the left atrium and baffle redirection of the right upper lobe vein. PMID- 9215229 TI - Remnants of the right valve of the sinus venosus presenting as a right atrial mass on transthoracic echocardiography. AB - Three patients were referred for suspicion of intracardiac tumour on transthoracic echocardiography. In all patients, the mass appeared as a nonobstructive oval structure measuring approximately 12 x 4 mm, located near the posterior third of the interatrial septum in the right atrium in the apical four chamber view. The characteristics of the mass were not those of a Eustachian valve or a Chiari network. Multiplane transesophageal echocardiography performed in each of these patients did not reveal a tumour but rather a fibrous band in the right atrium, extending from the inferior to the superior vena cava. These findings are consistent with remnants of the right valve of the sinus venosus. Inclusion of a persistent right valve of the sinus venosus in the differential diagnosis of a right atrial mass can alleviate concern and spare an unnecessary transesophageal examination when the typical transthoracic echocardiographic characteristics are identified. PMID- 9215231 TI - Time-dependent change in fast pathway refractoriness after slow pathway ablation in atrioventricular node reentrant tachycardia. Mansfield Polaris Investigators. AB - OBJECTIVE: To determine the time course of change in fast pathway refractoriness after slow pathway ablation. BACKGROUND: Antegrade fast pathway refractoriness has been observed to shorten in patients undergoing slow pathway ablation for atrioventricular (AV) node reentrant tachycardia. The time course and mechanism of this observation have not been explained. METHODS: Twenty-eight patients with AV node reentrant tachycardia and dual AV node pathways undergoing slow pathway ablation had the fast pathway effective refractory period (ERP) assessed immediately before, and at 0, 15, 30 and 45 mins after slow pathway ablation (Group 1). Twenty-five additional patients with AV node reentry and dual pathways involved in a multicentre protocol evaluating the Mansfield Polaris LE catheter underwent assessment of fast pathway refractoriness before and after slow pathway ablation, and at a routine three-month follow-up electrophysiology study (Group 2). RESULTS: In Group 1, antegrade fast pathway ERP fell from 394 ms before ablation to 334 ms immediately after slow pathway ablation, increased to 348 ms within 15 mins and was 353 ms at 45 mins (ANOVA P < 0.001). Retrograde fast pathway ERP fell from 325 ms before ablation to 294 ms at 45 mins (P = 0.02). In Group 2, antegrade fast pathway ERP fell from 390 ms before ablation to 337 ms after ablation, and rose to 362 ms at three months (P = 0.01). Retrograde fast pathway ERP also fell from 347 ms to 319 ms after ablation (P = 0.01), and remained unchanged at three months. CONCLUSION: Slow pathway ablation results in an immediate and sustained change in antegrade and retrograde first pathway refractoriness. There are immediate reversible and long term nonreversible components to this phenomenon. The latter finding may be related to loss of electrotonic inhibition of the fast pathway by the slow pathway. PMID- 9215230 TI - Muscle characteristics in effort angina before and after CABG. AB - Seven males with effort angina undertook graded ergometer tests and had muscle biopsies taken from their vastus lateralis muscle before, and three and six months after coronary bypass surgery. Muscle fibre composition (percentage of slow twitch fibres), ubiquinone (vitamin Q), and oxidative and fermentative enzyme activities were determined. After six months, muscle ubiquinone and oxidative enzymes were still depressed, indicating sustained muscle trauma. The only peripheral changes were that muscle lactate dehydrogenase and its skeletal muscle-specific subunits and isozymes were increased 35% to 40% (P < 0.001) three to six months postsurgery. Onset of blood lactate accumulation (2.0 mmol/L), symptom-limited ('maximal') exercise and peak blood lactate increased linearly over time (r = 0.52, P < 0.05; r = 0.63, P < 0.01; and r = 0.76, P < 0.001, respectively). It is suggested that the initial physical performance increase was due to improved circulatory capacity, oxygen delivery and lactate efflux, whereas the increased fermentative capacity ('anaerobic power') first contributed after a lag of three or more months. Whether the muscle histochemical changes reflected a healing process (recovery) is speculative. PMID- 9215232 TI - Rationale and design features of a clinical trial examining the effects of cholesterol lowering and angiotensin-converting enzyme inhibition on coronary atherosclerosis: Simvastatin/Enalapril Coronary Atherosclerosis Trial (SCAT). SCAT Investigators. AB - BACKGROUND: In the treatment of coronary atherosclerotic artery disease (CAD), the mechanisms by which lipid lowering, a proven therapy, produces beneficial clinical effects remain unclear. Moreover, although potential mechanisms of benefit are well known and increasingly applied clinically, there are no conclusive data from clinical trials studying primarily the antiischemic effects of angiotensin-converting enzyme (ACE) inhibition in patients with normal heart function. The Simvastatin/Enalapril Coronary Atherosclerosis Trial (SCAT) is designed to clarify some of these issues in CAD patients with normal or mildly elevated cholesterol. DESIGN AND OBJECTIVES: SCAT is a three- to five-year, multicentre, randomized, double-blind, placebo controlled, 2 x 2 factorial trial evaluating the effects of cholesterol lowering therapy by simvastatin and/or ACE inhibition by enalapril on anatomic coronary atherosclerosis progression assessed by quantitative coronary angiography in CAD patients with preserved left ventricular function and total cholesterol levels between 4.1 and 6.2 mmol/L. PATIENTS: Of 460 patients (age 61 +/- 9 years; 409 males, 51 females) enrolled between June 1991 and July 1995, 230 were randomized to simvastatin and 230 to placebo, and 229 to enalapril and 231 to placebo. Average baseline total cholesterol level was 5.20 +/- 0.61 mmol/L, high density lipoprotein cholesterol was 0.99 +/- 0.25 mmol/L, low density lipoprotein cholesterol was 3.36 +/- 0.57 mmol/L and triglycerides were 1.82 +/- 0.75 mmol/L. The trial will be completed in June 1998. SIGNIFICANCE: Insights gained from this long term angiographic trial will lead to a better understanding of the mechanisms of benefits of these two treatments, both alone and in combination. Of particular interest is that this trial will be able to examine a suspected beneficial interaction, if present, between these two treatments. PMID- 9215233 TI - Atrioventricular pacing does not increase infarct size in the in situ rabbit heart. AB - OBJECTIVE: To examine the hypothesis that an altered left ventricular depolarization sequence may augment infarct size. METHODS: Twenty-one New Zealand male rabbits were anesthetized and ventilated. The chest was opened and two electrodes were placed on the right atrium and ventricle. The rabbits were then randomized to atrial (n = 7), atrioventricular (AV) sequential (n = 7) or no (n = 7) pacing. The pacing rate was 20 beats/min higher than the sinus rate. After 1 min of pacing, the left coronary artery was occluded by a snare. After 30 mins, the snare was released and pacing was stopped. After 120 mins of reperfusion the experiment was terminated. Normal areas and areas at risk were delineated, infarct size was measured and the infarcted areas and areas at risk were planimetered. RESULTS: All results were expressed in cubic centimetres, and the ratio of the infarcted area to area at risk was calculated as a percentage (%I:R). The double product during ischemia was 21,546 +/- 2300 in controls, 23,000 +/- 3005 in rabbits with atrial pacing and 24,418 +/- 4253 in rabbits with AV pacing (F = 1.33, P = 0.28), and %I:R was 41.4 +/- 19.8, 43.9 +/- 15.4 and 38.4 +/- 18.4 (F = 0.16, P = 0.84), respectively. CONCLUSIONS: An altered left ventricular depolarization sequence in rabbit hearts does not increase infarct size. PMID- 9215234 TI - Compression of a composite graft by a peri-aortic pseudoaneurysm in a postoperative Marfan patient. AB - A previously well 35-year-old female was referred to hospital with a four-week history of intermittent chest pain. She had a history of Marfan syndrome when she presented 11 years previously with aortic insufficiency and underwent replacement of the aortic root with a composite graft. Magnetic resonance examination of the chest and coronary angiography revealed an area of extraluminal flow posteromedially at the base of the aortic graft with no evidence of dissection. Transesophageal echocardiography clearly demonstrated the presence of a pseudoaneurysm that resulted from detachment of the left coronary artery from the graft, with intermittent compression of the aortic graft during systole. The patient underwent uneventful resection and replacement of the false aneurysm and patch repair of the left coronary artery. This unusual case illustrates the presence of a pseudoaneurysm as a result of detachment of the left coronary artery from a Dacron graft 11 years after a composite graft replacement. PMID- 9215235 TI - Electrocardiographic abnormalities in right ventricular infarction associated with right bundle branch block. AB - An 80-year-old woman with pre-existing complete right bundle branch block presented with severe chest pain. The 12-lead electrocardiogram, together with right-sided chest leads, showed complete right bundle branch block and ST segment elevation in leads II, III, aVF, V5, V6 and V4R to V6R. These electrocardiographic abnormalities indicate acute 'Q wave' inferolateral and right ventricular infarction coexisting with right bundle branch block. PMID- 9215237 TI - Allergy care in the next millennium: guidelines for the specialty. PMID- 9215236 TI - New frontiers for IL-5. PMID- 9215238 TI - How should inhaled glucocorticoids be compared? PMID- 9215239 TI - Mite-contaminated foods as a cause of anaphylaxis. AB - BACKGROUND: Although insect and arthropod contamination of certain foods has been recognized for many years, allergic manifestations caused by ingestion of mite allergens have only rarely been reported. OBJECTIVE: The purpose of this study is to present clinical observations in patients who experienced acute anaphylaxis after eating mite-contaminated foods. METHODS: Thirty atopic subjects who were first seen with systemic anaphylaxis precipitated by the ingestion of wheat containing foods underwent skin prick tests with inhalant and food extracts, as well as with uncontaminated and mite-contaminated wheat flour. Flour samples were examined microscopically for identification and counting of mites. Der p 1 and Der f 1 levels were quantitated by using immunochemical methods. RESULTS: The most common symptoms were breathlessness, angioedema, wheezing, and rhinorrhea, which started between 10 and 240 minutes after eating. Abundant mites were present in the flour obtained from 28 patients; Suidasia spp. mites were found in grated bread from the other two patients. Positive prick test responses to Dermatophagoides farinae-and mite-contaminated flour and negative skin test responses to wheat extract, other food extracts, and uncontaminated wheat flour were found in all patients. Skin test responses were positive in volunteers with mite allergy even after heating the mite-contaminated flour at 100 degrees C. Screening of 35 unselected flour samples demonstrated the presence of mites in 13 of them (37.1%). CONCLUSIONS: Systemic anaphylaxis can occur after the ingestion of heated or unheated mite-contaminated foods. This problem may be more prevalent in tropical and subtropical countries than previously recognized. PMID- 9215240 TI - Treatment of anaphylactic sensitivity to peanuts by immunotherapy with injections of aqueous peanut extract. AB - BACKGROUND: Immediate hypersensitivity to peanuts is a frequent cause of anaphylactic reactions and deaths in children and adults. Currently, preventive treatment consists of avoidance, which is difficult because of the widespread and often disguised use of peanuts in the food industry. METHODS: Twelve patients with immediate hypersensitivity to ingestion of peanuts were recruited. Half were treated with injections of peanut extract: a maintenance level of tolerance was first achieved by a rush protocol, then maintained with weekly injections for at least 1 year. The other six were untreated control subjects. All patients underwent double-blind, placebo-controlled, oral peanut challenges initially, after approximately 6 weeks, and after 1 year. RESULTS: All treated patients achieved the maintenance dose of 0.5 ml of 1:100 wt/vol peanut extract by the rush injection protocol. All experienced increased tolerance to double-blind, placebo-controlled peanut challenge and decreased sensitivity on titrated skin prick testing with peanut extract, whereas the threshold to oral peanut challenge and cutaneous reactivity to peanut extract were unchanged in the untreated control subjects. Systemic reactions were common in the treated group both during rush immunotherapy and with maintenance injections. Only three patients remained tolerant of the full maintenance dose. The increased tolerance to oral peanut challenge was maintained in the three subjects who received full maintenance doses, but there was partial (n = 2) or complete (n = 1) loss of protection in the patients who required dose reduction because of systemic reactions. CONCLUSIONS: Injections of peanut extract increase the tolerance of patients with peanut allergy to oral ingestion of peanuts. Injections result in repeated systemic reactions in most patients, even during maintenance injections. For clinical application of this method of treatment, a modified peanut extract is needed. PMID- 9215241 TI - High levels of eosinophil cationic protein in wheezing infants predict the development of asthma. AB - BACKGROUND: In association with respiratory tract infections, infants may have episodes of wheezing, which represent the onset of asthma in some of them. Activated eosinophils play a central part in asthmatic inflammation. OBJECTIVE: We investigated whether, in infants experiencing their first episode of wheezing, eosinophil activation is present and can predict the development of asthma. METHODS: In a prospective trial, eosinophil activation was measured by eosinophil cationic protein (ECP) concentrations in serum from 33 nonatopic infants with their first episode of wheezing, 15 nonatopic infants with upper respiratory tract infection without wheezing, and 18 healthy nonatopic infants. One year later, the children were re-evaluated for a diagnosis of infantile asthma. RESULTS: Wheezing infants had higher median serum ECP levels (13.4 micrograms/L) than children with nonwheezy respiratory tract infection (7.6 micrograms/L, p < 0.005) or healthy subjects (7.1 micrograms/L, p < 0.005). In addition, wheezing infants (n = 13) with serum ECP concentrations greater than 20 micrograms/L were more likely to have asthma within 1 year than patients with ECP levels less than 20 micrograms/L (odds ratio = 12.4; confidence interval, 4.6-33.5). CONCLUSION: Eosinophil activation measured by serum ECP is present in infants with their first episode of wheezing illness, especially in those infants in whom asthma subsequently develops within 1 year. These data may indicate a predictive value of serum ECP measurements in children with wheezing to identify those patients in whom infantile asthma is developing. These findings probably also indicate that serum ECP may be used to identify the children who need early antiinflammatory treatment. PMID- 9215242 TI - The monosodium glutamate symptom complex: assessment in a double-blind, placebo controlled, randomized study. AB - BACKGROUND: Considerable debate swirls about the validity of symptoms described by many people after ingestion of monosodium glutamate (MSG), and the question has remained unresolved largely because of a paucity of well-designed challenge studies. METHODS: We conducted oral challenge studies in self-identified MSG sensitive subjects to determine whether they had a statistically significant difference in the incidence of their specific symptoms after ingestion of MSG compared with placebo. First, 5 gm MSG or placebo was administered in random sequence in a double-blind fashion. Subjects who reacted only to a single test agent then underwent rechallenge in random sequence in a double-blind fashion with placebo and 1.25, 2.5, and 5 gm MSG. A positive response to challenge was defined as the reproduction of > of 2 of the specific symptoms in a subject ascertained on prechallenge interview. RESULTS: Sixty-one subjects entered the study. On initial challenge, 18 (29.5%) responded to neither MSG nor placebo, 6 (9.8%) to both, 15 (24.6%) to placebo, and 22 (36.1%) to MSG (p = 0.324). Total and average severity of symptoms after ingestion of MSG (374 and 80) were greater than respective values after placebo ingestion (232 and 56; p = 0.026 and 0.018, respectively). Rechallenge revealed an apparent threshold dose for reactivity of 2.5 gm MSG. Headache (p < 0.023), muscle tightness (p < 0.004), numbness/tingling (p < 0.007), general weakness (p < 0.040), and flushing (p < 0.016) occurred more frequently after MSG than placebo ingestion. CONCLUSIONS: Oral challenge with MSG reproduced symptoms in alleged sensitive persons. The mechanism of the reaction remains unknown, but symptom characteristics do not support an IgE-mediated mechanism. According to Food and Drug Administration recommendations, the symptoms, originally called the Chinese restaurant syndrome, are better referred to as the MSG symptom complex. PMID- 9215243 TI - Indoor allergen exposure is a risk factor for sensitization during the first three years of life. AB - BACKGROUND: The purpose of the study was to investigate the influence of environmental allergen exposure on allergic sensitization in infancy and early childhood. METHODS: A cohort of 1314 newborns was recruited and followed up prospectively at the ages 12, 24, and 36 months. The levels of major mite (Der p 1 and Der f 1) and cat (Fel d 1) allergens were determined from domestic carpet dust samples by sandwich ELISA. Specific serum IgE antibodies to mite and cat allergens were determined by CAP fluoroimmunoassay (Pharmacia). Logistic regression was used to assess the effects of allergen exposure, age, family history, and cord blood IgE simultaneously on the risk of sensitization. RESULTS: Children, who had been found to be sensitized at least once during the first 3 years of life, were found to be exposed to significantly higher house dust mite (median, 868 ng/gm vs 210 ng/mg; p = 0.001) and cat (median, 150 ng/gm vs 64 ng/gm; p = 0.011) allergen concentrations in domestic carpet dust compared with the group without sensitization. In homes with low (< or = 25th percentile) dust concentrations, the risk of sensitization to mite (1.6%), and cat (2.0%) is low, compared with 6.5% for mite and 6.3% for cat if the domestic exposure is above the 75th percentile. The dose-response relationships between allergen levels and sensitization indicate that the increase in sensitization risk at low allergen levels is more pronounced in cat allergy (p = 0.002) than in mite allergy (p = 0.026). In the group with a positive family history, lower mite and cat allergen concentrations are needed to achieve specific sensitization compared with the group with a negative family history. CONCLUSION: Our data indicate that avoidance measures in the domestic environment aimed at the primary prevention of allergen-driven sensitization should be introduced at the earliest possible stage, if possible during infancy. PMID- 9215244 TI - Candida endocarditis in a child with hyperimmunoglobulinemia E syndrome. AB - Hyperimmunoglobulin E syndrome (HIE) is a disorder characterized by extremely elevated serum levels of IgE and recurrent infections. Patients are particularly predisposed to have staphylococcal abscesses, usually involving skin, lungs, and joints; but they are also at risk for infections with other bacteria and fungi. We report the case of a 46-month-old boy with HIE who had Candida endocarditis and sepsis with a large fungal mass extending through the tricuspid valve and into the surrounding heart tissue, requiring surgical excision and replacement with a prosthetic valve. He had an indwelling central line for previous antibiotic therapy and had oral thrush for a month before presentation, which had been treated with oral nystatin. He was first seen with very dark urine, a new murmur, petechial rash, in shock, and disseminated intravascular coagulation. The white blood cell count was 38,700 with 70% segmented neutrophils, 9% banded neutrophils, 15% lymphocytes, 4% monocytes, and 2% eosinophils. Hemoglobin was 7.1, and platelet count was 14,000. Prothrombin time was 15.5, and partial thromboplastin time was 31; fibrinogen level was 110 mg/ml, and fibrin degradation products were greater than 40 mg/ml. Serum IgE was 38,664 and 44,510 on repeat measurement. He has had recurrent staphylococcal pneumonias with pneumatoceles, twice requiring segmental lung resection. Blood and tricuspid valve cultures grew Candida albicans. He was treated with amphotericin and flucytosine, and later switched to fluconazole, with good response to therapy. A literature search revealed no other reported case of Candida endocarditis in patients with HIE. Fungai endocarditis is a rare complication, which may occur in patients with HIE and indwelling central catheters. PMID- 9215245 TI - A randomized, double-blind dose reduction study to compare the minimal effective dose of budesonide Turbuhaler and fluticasone propionate Diskhaler. AB - BACKGROUND: New inhaled glucocorticosteroids and inhalers are being developed. Their clinical equipotency is difficult to assess and is often discussed. OBJECTIVE: This study was carried out to compare the effect of budesonide Turbuhaler and fluticasone propionate (FP) Diskhaler in a dose reduction study in children (ages 5 to 16 years) with asthma. METHODS: Children treated with budesonide administered through a pressurized metered-dose inhaler with a large volume spacer had their budesonide dose gradually reduced to define the minimal effective dose with this delivery system. After this period, 217 children were randomly allocated to treatment with half the dose of either budesonide Turbukaler or FP Diskhaler for 5 weeks in a double-blind trial. If no deterioration in asthma control was seen, the dose was further reduced by 50% at 5-week intervals until deterioration in asthma control was seen. Throughout the study, morning and evening peak expiratory flow, symptoms, and use of rescue beta 2-agonist were recorded in diaries. Lung function tests and a standardized exercise test were performed at the clinic at the end of each treatment period. Urine cortisol excretion (24 hours) was measured before and after the first 5 week treatment period. Standardized criteria for deterioration in asthma control, based on diary card variables and exercise testing, were used to determine the minimal effective dose for each patient; and from this, the number of dose reduction steps was calculated. RESULTS: No statistically significant difference was seen in number of dose reduction steps from baseline or in minimal effective dose between the two treatments; mean reduction was 1.59 dose steps for budesonide Turbuhaler and 1.65 dose steps for FP Diskhaler (p = 0.52), and minimal effective dose was 188 micrograms for budesonide Turbuhaler and 180 micrograms for FP Diskhaler. After these dose reductions, the same level of asthma control was observed in the budesonide Turbuhaler and FP Diskhaler groups. Furthermore, no statistically significant differences between the two inhaler drug combinations were seen in daytime or nighttime symptoms, morning and evening peak expiratory flow, use of rescue beta 2-agonist, lung functions at the clinic, exercise-induced fall in lung function, or 24-hour urinary cortisol excretion during the first 5-week period. CONCLUSION: Microgram for microgram, budesonide Turbuhaler and FP Diskhaler are equally effective in treatment of children with moderate asthma. PMID- 9215246 TI - Mapping of IgE-binding epitopes on the recombinant major group I allergen of velvet grass pollen, rHol l 1. AB - BACKGROUND: New and more successful approaches to diagnosis and therapy of allergic diseases require a more subtle understanding of the structure and the epitopes on the allergen molecule. OBJECTIVE: This study was done to obtain more information on the structure and the IgE-binding epitopes of a major allergen of velvet grass pollen, Hol l 1. METHODS: We cloned Hol l 1 from a complementary DNA library and performed B-cell epitope mapping with 21 recombinant fragments expressed as fusion proteins in Escherichia coli. The fragments were analyzed by Western blotting with sera from 50 different patients. RESULTS: The patients' sera individually recognized at least four different IgE-binding regions (amino acids 1 to 27, 61 to 76, 84 to 105, and 158 to 240). According to their binding patterns with these epitopes, they were divided into five groups. Most sera (92%) bound to the C-terminal peptide (158 to 240), which consists of more than 80 amino acids, whereas there was virtually no binding to smaller fragments covering this region. In contrast to the C-terminal peptide, the IgE-binding peptides on the N terminus and on the middle region of the molecule were of a smaller size (15 to 30 amino acids). CONCLUSIONS: The major group I allergen of velvet grass bears at least four different IgE-binding epitopes, which were individually recognized by sera from different patients. The C terminus represents the major IgE-binding region and contains at least one discontinuous IgE-binding epitope, whereas the N terminus and middle region of Hol l 1 seem to contain continuous IgE-binding epitopes. PMID- 9215247 TI - Depletion of circulating allergen-specific TH2 T lymphocytes after allergen exposure in asthma. AB - BACKGROUND: In allergic asthma, CD4+ T lymphocytes are a fundamental component of local chronic inflammation. Their cytokine profile is oriented toward a TH2 phenotype, characterized by production of IL-4, IL-5, IL-10, and IL-13. Egress of T cells from blood to airways after allergen challenge has been described. OBJECTIVE: We have studied a cohort of six patients with asthma who had multiple allergies to investigate how exposure to allergen affects the proliferation of peripheral CD4+ T lymphocytes with different allergen specificities and lymphokine profiles. METHODS: For each patient, CD4+ T-cell lines were generated by in vitro stimulation with sensitizing and with nonsensitizing allergens, and IL-4 and interferon-gamma production by these lines was assessed. Proliferation of peripheral blood CD4+ T lymphocytes in response to the same allergens was measured before and 24 hours after inhalation challenge with a sensitizing allergen. RESULTS: We found that each single sensitizing allergen can deplete peripheral blood of TH2-type CD4+ T lymphocytes specific for all sensitizing allergens, but not of TH1-type CD4+ T lymphocytes. CONCLUSIONS: Our results suggest the existence of mechanisms capable of sorting disease-associated antigen specificities together with defined lymphokine patterns into T lymphocytes that can migrate to target organs, in allergic asthma. PMID- 9215248 TI - Determinants of airway responses to cat allergen: comparison of environmental challenge to quantitative nasal and bronchial allergen challenge. AB - BACKGROUND: Why allergic subjects may have asthma or rhinitis off allergen exposure remains unclear. OBJECTIVE: This study was carried out to compare airway responses during environmental allergen challenge (EAC) with quantitative allergen provocation challenges of the upper and lower airways. METHODS: Thirteen subjects with allergy to cats underwent EAC to cats. Lower airway responses during EAC were compared with bronchoprovocation with allergen. Nasal mucosal challenge with allergen-soaked disks were compared with EAC nasal responses. Nonspecific bronchial reactivity was assessed with methacholine; allergen sensitivity was assessed by skin prick tests, RAST, and end-point skin titration. RESULTS: During EAC, the maximal fall in FEV1 ranged from 6% to 57% (median, 18%) and correlated closely with allergen bronchoprovocation PD20 (Spearman's correlation coefficient [Rs] = -0.85, p < 0.0002). EAC asthmatic responses and allergen bronchoprovocation correlated with methacholine PD20 (Rs = -0.85, p = 0.0002 and Rs = 0.83, p = 0.0004, respectively). Nasal provocation and EAC nasal responses correlated with each other but not with lower airway responses. On the basis of EAC and allergen bronchoprovocation responses, seven participants with asthma were identified. This group was significantly more sensitive to inhaled methacholine but was similar to the nonasthmatic group in IgE-mediated sensitivity and nasal responses. CONCLUSIONS: The lower respiratory tract is less responsive to allergic and nonallergic stimuli in persons with allergic rhinitis. In persons with asthma during EAC, the response to nebulized cat allergen is also abnormal and correlates closely with their abnormal responsiveness to nonimmunologic stimuli. PMID- 9215250 TI - Diamine oxidase-gold enzyme-affinity ultrastructural demonstration that human gut mucosal mast cells secrete histamine by piecemeal degranulation in vivo. AB - Biopsy specimens of human ilea were prepared for the ultrastructural detection of histamine in vivo with a new enzyme-affinity method (Dvorak et al., J Histochem Cytochem 1993; 41:787-800). Human gut mucosal mast cells contained histamine in cytoplasmic secretory granules that were electron dense, regardless of underlying substructural patterns. Partially and completely empty cytoplasmic granules, characteristic for piecemeal degranulation (Dvorak et al., Int Arch Allergy Immunol 1992;99:74-83) had diminished and absent histamine stores, whereas electron-dense granules in the same cells retained their histamine. Interstitial collagen (but not basal lamina of other cells) near secretory mast cells avidly bound histamine, thereby providing a potential source for re-release of extracellular histamine stores. These studies provide the first documentation of histamine secretion in vivo during piecemeal degranulation of human mast cells. PMID- 9215249 TI - Cellular inflammatory responses and mediator release during early developing late phase allergic cutaneous inflammatory responses: effects of cetirizine. AB - BACKGROUND: Events in developing cutaneous late-phase allergic reactions can be characterized by a combination of skin chamber and biopsy approaches. In some previous studies, cetirizine reportedly inhibited mediator release and/or inflammatory cell responses in late-phase reactions. OBJECTIVE: This study was carried out to determine the effects of cetirizine on early late-phase reactions by using skin chamber and skin biopsy specimens. METHODS: Skin chamber responses during a 6-hour challenge with pollen antigens were assessed in 15 sensitive subjects during randomized, crossover treatment with cetirizine (20 mg/day) or placebo for 7-day periods with measurements of humoral and cellular components. Biopsy specimens of the underlying dermis were obtained. RESULTS: During cetirizine treatment, there was significant (p < 0.01) inhibition of immediate wheal and flare reactions to pollen antigens (34, 46%), codeine (41, 65%), and histamine (38, 68%). However, gross late-phase reactions at 6 hours were unaffected. During both cetirizine and placebo treatment, there was significantly greater accumulation at antigen sites in: (1) skin chamber levels of histamine, total cells, lactoferrin, and eosinophil cationic protein; (2) eosinophils (total and activated) on appended cover glasses; (3) deposition of lactoferrin and eosinophil cationic protein in the underlying dermis. However, these responses were not significantly different during cetirizine treatment compared with placebo treatment periods. CONCLUSION: A persistent pattern of inflammatory cell accumulation with release of granule proteins during early late-phase reactions was unaffected by cetirizine treatment. PMID- 9215251 TI - Purification of a major allergen, Asp f 2 binding to IgE in allergic bronchopulmonary aspergillosis, from culture filtrate of Aspergillus fumigatus. AB - BACKGROUND: Most cases of allergic bronchopulmonary aspergillosis (ABPA) are caused by the fungus Aspergillus fumigatus. Successful treatment of this disease depends on early diagnosis with the use of well-characterized and relevant antigens/allergens of the organism. OBJECTIVE: The aim of this study was to purify and characterize relevant proteins from A. fumigatus that could be used in the reliable diagnosis of ABPA. METHODS: Monoclonal antibodies were raised against A. fumigatus culture filtrate antigens. A Concanavalin A nonbinding protein fraction was purified with use of one of the monoclonal antibody immunoaffinity columns. The purified protein was analyzed on sodium dodecylsulfate-polyacrylamide gel electrophoresis gel and Western blots. The sensitivity and specificity of the purified protein were evaluated by RAST and ELISA with sera from 25 patients with ABPA, from 10 with allergic asthma, and from 10 normal control subjects. RESULTS: The 37 kD Concanavalin A nonbinding protein reacted specifically with IgE antibodies in patients with ABPA. Among the 25 patients with ABPA studied, 96% had IgE antibody against the allergen, whereas none of the subjects with allergic asthma who had positive results on the skin prick test or normal control subjects had a reaction. Both RAST and ELISA results exhibited strong correlation with IgE binding. This allergen exhibited N-terminal sequence identity to a recombinant allergen Asp f 2. CONCLUSIONS: A 37 kD protein with complete N-terminal homology to Asp f 2 is a major allergen of A. fumigatus that significantly reacts with IgE antibody in patients with ABPA, but does not elicit reaction in Aspergillus-sensitive subjects with asthma and normal control subjects. PMID- 9215253 TI - IL-5 synthesis is upregulated in human nasal polyp tissue. AB - BACKGROUND: In most nasal polyps, tissue eosinophilia is a striking finding, the pathologic mechanism of which is not understood. OBJECTIVE: This study was performed to investigate a possibly distinct cytokine and chemokine pattern that could explain the characteristic tissue eosinophilia in nasal polyps. METHODS: Polyps from 23 patients and turbinate tissue from 18 control subjects were investigated. The cytokine protein content (IL-1 beta, IL-3, IL-4, IL-5, IL-6, IL 8, IL-10, tumor necrosis factor-alpha, granulocyte-macrophage colony-stimulating factor, IL-1RA, RANTES, GRO-alpha) of tissue homogenates was measured by ELISA. Immunohistochemistry was performed in selected samples to detect IL-5+, major basic protein-positive, and EG2+ cells. RESULTS: IL-5 was detectable in only one sample of tissue from 18 control subjects but was found in 18 of 23 nasal polyps. Immunohistochemistry revealed an abundant number of IL-5+ cells, of which 69.5% could be identified as eosinophils by morphology. IL-6, IL-8, IL-10, tumor necrosis factor-alpha, GRO-alpha, and RANTES were detected in all specimens, without significant differences between groups (p > or = 0.05), whereas significnatly higher concentrations of IL-1 beta and IL-1RA were found in turbinate mucosa (p < or = 0.05). IL-3 was not detectable: granulocyte-macrophage colony-stimulating factor could only occasionally be found. CONCLUSION: This study indicates that IL-5 plays a key role in the pathophysiology of eosinophilic nasal polyps and may be produced by eosinophils. PMID- 9215252 TI - High serum IgE concentrations: association with HLA-DR and markers on chromosome 5q31 and chromosome 11q13. AB - BACKGROUND: Linkage studies mapped a locus regulating total serum IgE concentrations in a noncognate fashion to chromosome 5q31 and a locus for atopy to chromosome 11q13. In contrast, antigen-driven IgE production seems to be largely controlled by major histocompatibility complex class II genes. OBJECTIVE: We therefore analyzed the association between the phenotype of high IgE serum levels and six microsatellite markers on chromosomes 5q31 and 11q13, as well as HLA-DRB1, in a random sample of the adult East German population. METHODS: One hundred twenty-nine persons identified as "cases" (serum IgE level > 200 kU/L) and 266 control subjects (serum IgE level < or = 200 kU/L) were genotyped for five 5q31 microsatellites (D5S436, D5S393, D5S210, IL-4, and IL-9) and an 11q13 microsatellite (FCERIB). Cases and controls were also typed for HLA-DRB1. Allele frequencies were compared between cases and controls by means of a two-sided Fisher's exact test. RESULTS: None of the markers was significantly associated although a weak association to the markers within the IL-9 gene and the FCER1B gene and to the HLA-DRB1*01 allele was found when specific IgE-positive cases were compared with negative controls. CONCLUSIONS: The weak associations observed after stratification for specific IgE might point to a contribution of genes in these regions to the development of allergy. PMID- 9215254 TI - Vocal cord dysfunction with nocturnal awakening. PMID- 9215255 TI - Usual interstitial pneumonitis in a patient with common variable immunodeficiency. PMID- 9215256 TI - Allergic reactions to North African scorpion venom evaluated by skin test and specific IgE. PMID- 9215258 TI - Food allergy to Belgian endive (chicory). PMID- 9215257 TI - A comparison of drug delivery from a metered-dose inhaler plus an inspiratory flow control device with a metered-dose inhaler plus a spacer device. PMID- 9215259 TI - Cyclosporine restores cytokine imbalance in childhood atopic dermatitis. PMID- 9215260 TI - Alcohol-induced bronchial asthma. PMID- 9215261 TI - Alcohol-induced asthma: not only in Asians. PMID- 9215262 TI - Cetirizine overdose in a young child. PMID- 9215263 TI - Topical triclosan treatment of atopic dermatitis. PMID- 9215264 TI - Flovent (fluticasone propionate inhalation aerosol) PMID- 9215265 TI - The prevalence and medical and economic impact of allergic rhinitis in the United States. PMID- 9215266 TI - Clinical review 89: Small as fetus and short as child: from endogenous to exogenous growth hormone. PMID- 9215267 TI - Childhood adrenocortical tumors. PMID- 9215268 TI - Why retest young adults with childhood-onset growth hormone deficiency? PMID- 9215269 TI - Thyroid gland abnormalities in patients with the syndrome of spotty skin pigmentation, myxomas, endocrine overactivity, and schwannomas (Carney complex) AB - Carney complex is a multiple neoplasia and lentiginosis syndrome that affects endocrine glands, including the pituitary, adrenals, and testes; thyroid gland involvement has not been unequivocally demonstrated. In the present study, the medical records of 12 families with Carney complex (53 affected patients) were reviewed for evidence of thyroid abnormality; 2 patients with thyroid carcinoma (1 papillary and 1 follicular; 3.8%) and 1 with follicular adenoma were identified in 3 unrelated kindreds. Six affected members of these kindreds were then screened for the presence of thyroid disease (familial cases). We also studied 5 patients with the complex who had no affected relatives (sporadic cases). These 11 patients consisted of 5 adults [mean age, 33.2 +/- 9.2 (+/- SD) yr] and 6 children and adolescents (mean age, 13.8 +/- 2.5 yr). All had normal results of physical and biochemical examination of the thyroid gland (total and free T4, T3, and TSH levels). Thyroid ultrasonography showed hypoechoic, cystic, solid, or mixed lesions in 3 of the 5 adults (60%) and 4 of the 6 children (67%). Two patients underwent fine needle aspiration biopsy, which identified follicular lesions. Thyroid gland abnormalities were documented in 5 siblings and 1 parent child pair. We conclude that thyroid gland pathology is 1) common in patients with Carney complex; 2) includes a spectrum of abnormalities ranging from follicular hyperplasia and/or cystic changes to carcinoma; and 3) is inherited in an autosomal dominant manner, like the other manifestations of the syndrome, it is therefore, a candidate component of the syndrome. Ultrasonography is useful in the detection and clinical follow-up of these lesions. PMID- 9215270 TI - Long-term testosterone administration increases visceral fat in female to male transsexuals. AB - The amount of intraabdominal (visceral) fat is an important determinant of disturbances in lipid and glucose metabolism. Cross-sectional studies in women have found associations between high androgen levels and visceral fat accumulation. The causal relation between these phenomena is unknown. We, therefore, studied prospectively the effect of testosterone administration on body fat distribution in 10 young, nonobese, female to male transsexuals undergoing sex reassignment. Before, after 1 yr, and after 3 yr of testosterone administration, magnetic resonance images were obtained at the level of the abdomen, hip, and thigh to quantify both sc and visceral fat depots. After 1 yr of testosterone administration, sc fat depots at all levels showed significant reductions compared to baseline measurements. The mean visceral fat area did not change significantly, but subjects who gained weight in the first year after testosterone administration showed an increase in visceral fat. After 3 yr of testosterone administration, sc fat depots were no longer significantly lower compared to pretreatment measurements, but the mean visceral fat depot had increased significantly by 13 cm2 (95% confidence interval, 4-22 cm2), a relative increase of 47% (95% confidence interval, 8-91%) from baseline. The increase in visceral fat was most pronounced in those subjects who had gained weight. We conclude that long term testosterone administration in young, nonobese, female subjects increases the amount of visceral fat. In addition, an increase in weight in this hyperandrogenic state leads to a preferential storage of fat in the visceral depot. PMID- 9215271 TI - Regulation of intercellular adhesion molecule-1 expression in human thyroid cells in vitro and human thyroid tissue transplanted to the nude mouse in vivo: role of Graves' immunoglobulins and human thyrotropin receptor. AB - To further explore a potential role for the human TSH receptor (hTSHR) in the propagation of thyroid autoimmune disease, we examined immunomodulatory effects in response to its stimulation by Graves' Igs both in human thyroid tissue transplanted to the nude mouse and in primary cultures of human thyrocytes. We injected nude mice bearing transplants derived from normal human thyroid with protein A-Sepharose-purified Graves' IgGs (0.05-1 mg) on 2 days and assessed, in addition to functional stimulation, the expression of intercellular adhesion molecule-1 (ICAM-1) by transplant thyrocytes. In parallel to functional stimulation, as demonstrated by thyroid follicular cell hypertrophy in the transplants and increased T4 production, Graves' IgGs induced a marked dose dependent expression of ICAM-1 by transplanted thyrocytes, which exceeded that of a continuous interferon-gamma infusion (200 IU/24 h) for 2 days. Normal IgGs were ineffective. Bovine TSH (bTSH) had little effect by itself, but did enhance interferon-gamma-induced ICAM-1 expression. To assess the specificity of their effects, experiments with Graves' IgGs were conducted in the presence and absence of a selective hTSHR antagonist (asialoagalacto-hCG). Asialoagalacto-hCG nearly completely abolished the stimulatory effect of Graves' IgGs on ICAM-1 expression and significantly reduced the combined bTSH/interferon-gamma effect. It failed, however, to affect interferon-gamma action. In vitro studies using human thyroid cells in primary culture confirmed the in vivo observations, treatment with saline resulted in 14% of cells expressing ICAM-1, with pooled normal IgGs (500 mg/L) in 18% and with Graves' IgGs (patient A, 448 mg/L; patient B, 260 mg/L) in 78% and 51%, respectively. Upon exposure to Graves' IgGs (90 mg/L) plus asialo hCG(350 mg/L), 25% of the cells stained positively for ICAM-1, 29% to bTSH (10 IU/L), 31% to recombinant human TSH (10 IU/L), and 84% to interferon-gamma (10 IU/mL). In conclusion, stimulation of human thyroid cells, either transplanted to the nude mouse in vivo or studied under in vitro conditions, with Igs derived from patients with Graves' disease increased the expression of ICAM-1 on the surface of the cells. The action appears to be specific and mediated by the hTSHR. This particular property of TSHR autoantibodies may be of pathophysiological relevance in Graves' disease, as it may assist in targeting the autoimmune attack and in promoting lymphocyte recruitment to the thyroid gland. PMID- 9215272 TI - Serum bone alkaline phosphatase isoenzyme levels in normal children and children with growth hormone (GH) deficiency: a potential marker for bone formation and response to GH therapy. AB - Serum bone alkaline phosphatase (B-ALP) has been considered to be a good marker for bone formation. Recently, a specific immunoradiometric assay for serum B-ALP has been developed. Using this system, we measured the serum levels of B-ALP in 363 normal children (207 males and 156 females, age 0-18 yr) and in 20 GH deficient children (age 5-13 yr) who showed significant bone growth during GH therapy. We found the following results. 1) There were no significant circadian variations in serum B-ALP levels (coefficients of variation: 2.10-9.66%). 2) In normal children, serum B-ALP levels were high in infants and gradually declined and increased again during puberty. During the pubertal period, the highest serum B-ALP values were observed at midpuberty (stage 3 of breast and pubic hair development and 4-12 mL of testicular volume). 3) Serum B-ALP levels were significantly correlated with levels of the carboxy-terminal propeptide of type 1 procollagen (r = 0.447, P < 0.0001) and osteocalcin (r = 0.433, P < 0.0001). 4) After beginning GH therapy, serum B-ALP levels increased significantly; a 26% increase in serum B-ALP level was observed after 3 months of GH therapy. 5) The ratio between serum B-ALP level after 3 months of GH therapy and before GH therapy was positively correlated with the GH-induced improvement in the height SD score (height SD score after 1 yr of GH therapy minus that before GH therapy) and improvement in the height velocity SD score (height velocity SD score during GH therapy minus before GH therapy) (r = 0.531, P < 0.05 and r = 0.608, P < 0.01, respectively). 6) The increment of SD score in serum B-ALP level after 1 yr of GH treatment was also significantly correlated with that for bone mineral density after 1 yr of GH therapy (r = 0.663, P < 0.005). These results show that B-ALP levels are a useful marker for bone formation because B-ALP levels increased when the growth rate accelerated. Serum B-ALP is a potential predictor of the effectiveness of GH therapy, because the serum level after 3 months of GH therapy reflects the outcome of 1 yr of GH therapy. PMID- 9215273 TI - Disparate serum free testosterone concentrations and degrees of hypothalamo pituitary-luteinizing hormone suppression are achieved by continuous versus pulsatile intravenous androgen replacement in men: a clinical experimental model of ketoconazole-induced reversible hypoandrogenemia with controlled testosterone add-back. AB - To investigate the neuroendocrine mechanisms underlying the negative feedback actions of testosterone on both the pulsatile mode of LH release and the entropy or disorderliness of the LH release process, we blocked testicular androgen biosynthesis using oral high dose ketoconazole treatment with concomitant low dose glucocorticoid replacement for 48 h in six healthy young men. Volunteers were then infused iv with saline or a total of 8.0 mg testosterone base over the second 24 h via either a continuous or a pulsatile (90-min boluses) delivery pattern. Discrete peak detection (Cluster analysis) was applied to obtain a model independent estimate of the frequency of serum LH concentration peaks, maximal and incremental LH peak amplitudes, peak area, and interpeak nadir serum LH concentrations. Approximate entropy was used to quantify the relative orderliness/disorderliness of the LH release process over 24 h. Ketoconazole treatment markedly lowered 24-h mean serum total and free testosterone concentrations (by 17- and 9-fold respectively), and significantly increased LH pulse frequency, maximal LH peak height, and interpeak nadir serum LH concentrations. Continuous iv testosterone add-back increased 24-h pooled serum free testosterone concentrations 3-fold more and concomitantly reduced mean (24 h) serum LH concentrations by at least 2-fold more than pulsatile delivery of the same total daily amount of androgen. Both modes of testosterone infusion suppressed pulsatile LH release, but the effects were distinguishable; namely, treatment with continuous vs. intermittent androgen add-back, respectively, decreased LH pulse frequency and incremental LH pulse amplitude. Ketoconazole treatment alone also significantly increased approximate entropy values, indicating greater disorderliness of LH release during androgen removal. Approximate entropy/orderliness was restored to baseline by continuous, but not pulsatile, iv testosterone replacement. In conclusion, the present novel testosterone add-back clinical experimental paradigm indicates that 1) remarkably different 24-h mean serum free testosterone concentrations can result from continuous vs. pulsatile testosterone delivery into the bloodstream; 2) androgen negative feedback can exert frequency- as well as amplitude-dependent suppression of pulsatile LH release; and 3) testosterone is required to maintain an orderly 24-h LH release process in young men. PMID- 9215274 TI - Stress-induced declarative memory impairment in healthy elderly subjects: relationship to cortisol reactivity. AB - A group of 14 healthy elderly subjects was submitted to a nonstressful (attentional task) and a stressful (public speaking task) condition. Declarative (conscious recollection of learned information) and nondeclarative (retrieved information without conscious or explicit access) memory as well as salivary cortisol levels were measured before and after each condition. The results showed that the stressful condition significantly decreased declarative memory performance, whereas the nonstressful condition did not. Nondeclarative memory performance was not affected by either condition. Further analyses separating the subjects into responders and nonresponders in terms of stress-induced cortisol changes revealed a very different pattern of cortisol secretion and declarative memory performance in both populations. We showed that the responders presented increased cortisol levels 60 min before the actual stressor, whereas the nonresponders presented increased cortisol levels 25 min before the actual stressor. Although the responders did not differ from the nonresponders in declarative memory performance before and after the nonstressful condition, they presented a lower declarative memory performance when measured before and after the stressful condition. The early increase in cortisol levels observed in the responder group suggests that the anticipation of the stress, rather than the actual stressor per se, may have played a more significant role in the stress induced declarative memory deficits observed in this subgroup. Together, these results show that the cortisol response to anticipation of stress and/or to stress in the elderly specifically affects those memory functions that are dependent on hippocampal activity. They also suggest that an altered cortisol responsivity to acute and/or chronic stress, with its detrimental effects on memory, could be an important factor explaining the genesis of memory deficits in aged populations. PMID- 9215275 TI - Multiple endocrine abnormalities of the growth hormone and insulin-like growth factor axis in prepubertal children with exogenous obesity: effect of short- and long-term weight reduction. AB - We have studied the GH-insulin-like growth factor (IGF) axis in prepubertal children with exogenous obesity at the time of clinical diagnosis and at two time points during weight reduction on a calorie-restricted diet. Spontaneous GH secretion, IGF-I, free IGF-I (fIGF-I), IGF-II, their binding proteins (IGFBP-1, IGFBP-2, and IGFBP-3), and GH-binding protein (GHBP) values at the time of clinical diagnosis (n = 65), after a 25% decrease in the body mass index (BMI) expressed as the SD score (BMI SD score; n = 29), and after a diminution of at least 50% of the initial BMI SD score (n = 9) are reported. GH secretion was significantly reduced at diagnosis, and after a decrease of at least 25% in the initial BMI SD score, it returned to normal in all patients. Total IGF-I levels were not significantly different from those in controls at any point. In contrast, fIGF-1 and IGF-II levels were significantly increased, both at diagnosis and after BMI SD score reduction. Obese patients were hyperinsulinemic at diagnosis and remained so even after a 50% reduction of their BMI SD score. Serum IGFBP-1 and IGFBP-2 levels were significantly decreased at diagnosis and at the two points studied during weight reduction. Serum IGFBP-3 and GHBP levels were increased significantly at diagnosis and returned to normal levels after a reduction in the BMI SD score. A positive correlation between serum GHBP levels and BMI was found in both controls and obese patients. Serum IGFBP-3 levels correlated positively with IGF-I, fIGF-I, and IGF-II in all groups, but these correlations were weaker in the obese patients at diagnosis. IGFBP-2 correlated significantly with IGF-II only in the obese group at diagnosis (r = -0.760; P < 0.0001), but with fIGF-I in all groups. IGFBP-1 was negatively correlated with IGF-I and fIGF-I in all groups. In conclusion, the GH-IGF axis is dramatically altered in patients with exogenous obesity. However, most changes in the peripheral IGF system appear to be independent of the modifications in GH secretion. In addition, in contrast to current thought, not all of the observed abnormalities are reversed with a significant reduction in the BMI SD score. PMID- 9215277 TI - Dehydroepiandrosterone sulfate: a biomarker of primate aging slowed by calorie restriction. AB - The adrenal steroids, dehydroepiandrosterone (DHEA) and its sulfate (DHEAS), have attracted attention for their possible antiaging effects. DHEAS levels in humans decline markedly with age, suggesting the potential importance of this parameter as a biomarker of aging. Here we report that, as seen in humans, male and female rhesus monkeys exhibit a steady, age-related decline in serum DHEAS. This decline meets several criteria for a biomarker of aging, including cross-sectional and longitudinal linear decreases with age and significant stability of individual differences over time. In addition, the proportional age-related loss of DHEAS in rhesus monkeys is over twice the rate of decline observed in humans. Most important is the finding that, in rhesus monkeys, calorie restriction, which extends life span and retards aging in laboratory rodents, slows the postmaturational decline in serum DHEAS levels. This represents the first evidence that this nutritional intervention has the potential to alter aspects of postmaturational aging in a long-lived species. PMID- 9215276 TI - Multiple endocrine abnormalities of the growth hormone and insulin-like growth factor axis in patients with anorexia nervosa: effect of short- and long-term weight recuperation. AB - We have studied the GH-insulin-like growth factor (IGF) axis in patients with anorexia nervosa at the time of diagnosis and at two points during weight recuperation. We report their spontaneous GH secretion and IGF-I, free IGF-I (fIGF-I), IGF-II, the IGF-binding proteins (IGFBP-1, IGFBP-2, and IGFBP-3), and GH-binding protein (GHBP) levels at the time of the clinical diagnosis (n = 50) and after recuperation of between 6-8% (n = 42) and 10% or less of the initial weight (n = 20). Two distinct groups were seen, those who significantly hypersecreted GH and those whose GH secretion was reduced significantly. After recuperation of 10% or more of their initial weight, all patients had a normal GH pattern. Independently of GH secretory dynamics, IGF-1, IGFBP-3, and GHBP serum levels were all significantly decreased at diagnosis, and only GHBP returned to normal after weight recuperation. Serum IGFBP-1 and IGFBP-2 levels were significantly increased at the time of diagnosis and decreased after weight recuperation. The body mass index (BMI) correlated positively with fIGF-I levels and negatively with IGFBP-1 and IGFBP-2 levels, but only after weight recuperation in all cases. Contrary to what is seen in normal individuals, no correlation was found between BMI and serum GHBP levels in anorexia nervosa patients. Serum IGFBP-2 levels had a strong negative correlation with fIGF-I, IGF II, and the sum of IGF-I and IGF-II, but only at the time of diagnosis. In conclusion, the GH-IGF axis is dramatically altered in patients with anorexia nervosa. Changes in the peripheral IGF system however, appear to be independent of modifications in GH secretion and, in contrast to current thought, not all of the observed abnormalities are rapidly reversed with weight recuperation. PMID- 9215278 TI - Evidence for a heterozygote advantage in congenital adrenal hyperplasia due to 21 hydroxylase deficiency. AB - 21-Hydroxylase deficiency is one of the most common inherited disorders, with carrier frequencies of approximately 10% in all world populations studied to date. The high prevalence of the mutant gene is probably due to a flanking pseudogene serving as a reservoir for mutations. Despite the potential for a high rate of de novo mutations, a founder effect for specific gene conversions is observed in most populations. We hypothesized that there was a survival advantage to 21-hydroxylase heterozygotes, and here we report endocrinological and molecular investigations to test this hypothesis. We defined 28 carriers and 22 mutation-negative controls by molecular genotyping and determined ACTH-stimulated adrenal hormone responses. We found significantly elevated cortisol responses in the carriers compared to controls (30 min cortisol levels: normal, 24.2 +/- 4.6 micrograms/dL; carrier, 28.1 +/- 4.2 micrograms/dL; P < 0.005). Cortisol has a crucial role in maintaining homeostasis, influencing differentiation, suppressing inflammation, and effecting cross-talk among the immune, nervous, and endocrine systems. The brisk cortisol response we have documented in carriers of 21 hydroxylase may enable a rapid return to homeostasis in response to infectious, inflammatory, or other environmental stresses and may protect from inappropriate immune responses, such as autoimmune diseases. PMID- 9215279 TI - Sex-related difference in the growth of prolactinomas: a clinical and proliferation marker study. AB - Prolactinomas in women commonly present as small intrasellar tumors, but are usually much larger in men. This discrepancy has generally been attributed to differences in the delay before diagnosis. However, studies comparing clinical and pathological correlates of growth of these tumors in both sexes are lacking. We conducted a retrospective study comparing 45 men and 51 women bearing prolactinoma to determine whether the predominance of large tumors in men was due to a delay in diagnosis or, rather, to a fundamental sex-related difference in tumor growth. Basal PRL levels (mean +/- SEM, 2789 +/- 573 ng/mL) and mean tumor diameter (26 +/- 2 mm) were significantly higher in men than in women (292 +/- 74 ng/mL and 10 +/- 1 mm, respectively; P < 0.001), but were not correlated to the age at diagnosis or the duration of symptoms. Giant tumors (n = 8) occurred in males only. The frequencies of bromocriptine-resistant tumors (30 vs.5%; P < 0.01) and invasive macroadenomas (52 vs.27%; P < 0.001) were significantly greater in men than those in women. Lastly, macroprolactinomas in males exhibited higher indexes of proliferating cells by Ki-67 immunoreactivity (2.6 +/- 1.1% of positive nuclei) than did similar tumors in female patients (0.4 +/- 0.2%; P = 0.08). We conclude that the predominance of large prolactinomas in men is due to a high frequency of rapidly growing tumors, which are often invasive and frequently bromocriptine resistant. PMID- 9215281 TI - Growth hormone testing for the diagnosis of growth hormone deficiency in childhood: a population register-based study. AB - Evaluation of GH secretion using pharmacological GH stimulation tests (GHST) remains a current practice, although the reliability of GHST has been questioned, and many pitfalls have been pointed out. We have analyzed all of the 6373 GH stimulation tests that led to the initiation of GH therapy in 3233 children treated in France from 1973-1989. Tests and GH measurements were performed by individual centers and collected by the Association France-Hypophyse. GH deficiency (GHD) was due to craniospinal irradiation (11%), was due to organic causes or associated with multiple deficiencies (22%), or was considered idiopathic (65%); 2% of the patients were considered non-GHD. Eleven different pharmacological tests were used, and 62 of the 66 theoretical pairs of tests were used at least once. The most frequent combination of tests (ornithine in one instance and insulin in another) was used in 12.7% of patients. The reliability of the GH peak measured by comparing the results of 2 tests in the same patient was poor, as measured by intraclass correlation coefficients below 0.8. Multivariate analysis identified several parameters positively or negatively associated with peak plasma GH: calendar year of initiation of treatment, etiology of GHD, height SD score, bone age SD score, puberty, weight SD score, genetic target height SD score, and the nature of the pharmacological agent used. We believe that several of these factors (weight SD score, genetic target height SD score, and nature of the agent) identify biases in the diagnosis of GHD. We conclude that GHST should be performed with a very limited number of agents, interpreted after the establishment of reference values in age-matched normal children, and associated with other clinical and biochemical parameters for establishing the diagnosis of GHD. PMID- 9215280 TI - Troglitazone improves defects in insulin action, insulin secretion, ovarian steroidogenesis, and fibrinolysis in women with polycystic ovary syndrome. AB - Women with polycystic ovary syndrome (PCOS) are characterized by defects in insulin action, insulin secretion, ovarian steroidogenesis, and fibrinolysis. We administered the insulin-sensitizing agent troglitazone to 13 obese women with PCOS and impaired glucose tolerance to determine whether attenuation of hyperinsulinemia ameliorates these defects. All subjects had oligomenorrhea, hirsutism, polycystic ovaries, and hyperandrogenemia. Before and after treatment with troglitazone (400 mg daily for 12 weeks), all had 1) a GnRH agonist (leuprolide) test, 2) a 75-g oral glucose tolerance test, 3) a frequently sampled iv glucose tolerance test to determine the insulin sensitivity index and the acute insulin response to glucose, 4) an oscillatory glucose infusion to assess the ability of the beta-cell to entrain to glucose as quantitated by the normalized spectral power for the insulin secretion rate, and 5) measures of fibrinolytic capacity [plasminogen activator inhibitor type 1 (PAI-1) and tissue plasminogen activator]. There was no change in body mass index (39.9 +/- 1.4 vs. 40.2 +/- 1.4 kg/m2) or body fat distribution after treatment. Both the fasting (91 +/- 3 vs. 103 +/- 3 mg/dL; P < 0.001) and 2 h (146 +/- 8 vs. 171 +/- 6 mg/dL; P < 0.02) plasma glucose concentrations during the oral glucose tolerance test declined significantly. There was a concordant reduction in glycosylated hemoglobin to 5.7 +/- 0.1 from a pretreatment level of 6.1 +/- 0.1% (P < 0.03). Insulin sensitivity increased from 0.58 +/- 0.14 to 0.95 +/- 0.26 10(-5) min 1/pmol.L (P < 0.01) after treatment as did the disposition index (745 +/- 135 vs. 381 +/- 96; P < 0.05). The ability of the beta-cell to appropriately detect and respond to an oscillatory glucose infusion improved significantly after troglitazone treatment; the normalized spectral power for the insulin secretion rate increased to 5.9 +/- 1.1 from 4.3 +/- 0.8 (P < 0.05). Basal levels of total testosterone (109.3 +/- 15.2 vs. 79.4 +/- 9.8 ng/dL; P < 0.05) and free testosterone (33.3 +/- 4.0 vs. 21.2 +/- 2.6 pg/mL; P < 0.01) declined significantly after troglitazone treatment. Leuprolide-stimulated levels of 17 hydroxyprogesterone, androstenedione, and total testosterone were significantly lower posttreatment compared to pretreatment. The reduction in androgen levels occurred independently of any changes in gonadotropin levels. A decreased functional activity of PAI-1 in blood (from 12.7 +/- 2.8 to 6.3 +/- 1.4 AU/mL P < 0.05) was associated with a decreased concentration of PAI-1 protein (from 64.9 +/- 9.1 to 44.8 +/- 6.1 ng/mL; P < 0.05). No change in the functional activity of tissue plasminogen activator (from 5.3 +/- 0.4 to 5.1 +/- 0.5 IU/mL) was observed despite a decrease in its concentration (from 9.6 +/- 0.9 to 8.2 +/- 0.7 ng/mL; P < 0.05). The marked reduction in PAI-1 could be expected to improve the fibrinolytic response to thrombosis in these subjects. We conclude that administration of troglitazone to women with PCOS and impaired glucose tolerance ameliorates the metabolic and hormonal derangements characteristic of the syndrome. Troglitazone holds potential as a useful primary or adjunctive treatment for women with PCOS. PMID- 9215282 TI - Laminin suppresses progesterone production by human luteinizing granulosa cells via interaction with integrin alpha 6 beta 1. AB - We previously raised a murine monoclonal antibody (mAb), OG-1, against human granulosa cells (GC) and reported that human GC express the OG-1 antigen with the highest immunoreactivity during the periovulatory phase. Later, we showed that the OG-1 antigen is identical to human integrin alpha 6, and that human GC express integrin alpha 6 beta 1, but not alpha 6 beta 4. In the present study, we examined the expression of laminin (LN), the ligand for integrin alpha 6 beta 1. Flow cytometry showed that LN was bound to the cell surface of some GC obtained from preovulatory follicles of patients undergoing in vitro fertilization. Immunohistochemistry showed that LN was detected between luteinizing GC in the early corpora lutea. To examine the effect of LN on steroidogenesis by human luteinizing GC, GC obtained from patients undergoing in vitro fertilization were cultured on mouse LN-coated or noncoated plastic dishes in medium containing 5% FCS for 24 h. In the absence or presence in hCG (1 IU/mL), GC cultured on LN coated dishes produced 0.70- and 0.67-fold less progesterone than those on noncoated dishes, respectively (P < 0.05). We examined the effect of the interaction of integrin alpha 6 beta 1 and LN on steroidogenesis by human luteinizing GC. We cultured GC with 5 micrograms/mL of the anti-alpha 6 mAb GoH3, which inhibits the interaction between human integrin alpha 6 beta 1 and mouse LN, or with a control rat mAb (TER199) on mouse LN-coated dishes in serum-free medium for 24 h. In the absence or presence of hCG (1 IU/mL), GC cultured with GoH3 produced 1.97- and 1.94-fold more progesterone than the control cells (P < 0.01 and P < 0.05, respectively). In contrast, when GC were cultured on dishes coated with type IV collagen, progesterone production was not enhanced by GoH3. Furthermore, the anti-alpha 6 mAb OG-1, which does not inhibit the interaction between integrin alpha 6 beta 1 and LN, had no effect on the progesterone production by GC cultured on LN. These results indicate that LN suppresses the luteinization of human luteinizing GC via integrin alpha 6 beta 1 and that integrin alpha 6 beta 1 regulates the luteinization of human GC during the periovulatory phase. PMID- 9215283 TI - The human thyrotropin (TSH) receptor in a TSH binding inhibition assay for TSH receptor autoantibodies. AB - Seven years after the molecular cloning of the human TSH receptor (TSHR), the porcine TSHR remains in general use in the TSH binding inhibition (TBI) assay for autoantibodies to the TSHR. We compared porcine and recombinant human TSHR in two types of TBI assays: one using intact Chinese hamster ovary cells expressing the recombinant human TSHR on their surface, and the other using soluble receptors extracted from these cells with detergent. In the intact cell TBI assay, monolayers expressing large numbers of TSHR were less effective than cells expressing few receptors. These findings are consistent with the very low concentration of TSHR autoantibodies in serum. Binding of [125I]human TSH was about 5-fold lower than that of [125I]bovine TSH to the intact cells. Nevertheless, TBI values with the two ligands were similar for most sera. However, a few sera produced greater inhibition of human than of bovine TSH binding. In the solubilized human TSHR TBI assay, in contrast to the intact cell TBI assay, cells expressing very large number of TSHR were an excellent source for detergent extraction of soluble human TSHR, but only if the cells were extracted while still on the dish and not after scraping. A 10-cm diameter dish of cells provided TSHR for 100-200 replicate determinations when substituted for solubilized porcine TSHR in a commercial TBI kit. TBI values in serum from 30 individuals with suspected Graves' disease correlated closely when tested with solubilized human and porcine TSHR (r = 0.954; P < 0.001). However, 2 sera that were negative with the porcine TSHR were positive with the human TSHR. TBI and thyroid-stimulating activity in these sera correlated weakly regardless of whether the TBI used human or porcine TSHR. These findings open the way to a practical TBI assay using recombinant human TSHR. PMID- 9215284 TI - Follicle-stimulating hormone and luteinizing hormone/chorionic gonadotropin stimulation of vascular endothelial growth factor production by macaque granulosa cells from pre- and periovulatory follicles. AB - Granulosa cells in the ovulatory follicle express messenger ribonucleic acid encoding vascular endothelial growth factor (VEGF), an agent that may mediate the neovascularization of the developing corpus luteum, but it is not known whether luteinizing granulosa cells synthesize and secrete VEGF during the periovulatory interval. Studies were designed to evaluate the effects of an in vivo gonadotropin surge on VEGF production by macaque granulosa cells (study 1) and to test the hypothesis that gonadotropins act directly on granulosa cells to regulate VEGF production (study 2). Monkeys received a regimen of exogenous gonadotropins to promote the development of multiple preovulatory follicles. Nonluteinized granulosa cells (i.e. preovulatory; NLGC) and luteinized granulosa cells (i.e. periovulatory; LGC) were aspirated from follicles before and 27 h after an ovulatory gonadotropin bolus, respectively. Cells were either incubated for 24 h in medium with or without 100 ng/mL hCG (study 1) or cultured for 6 days in medium with or without 100 ng/mL hCG or 0.1, 1, 10, and 100 ng/mL of recombinant human LH (r-hLH) or r-hFSH (study 2). Culture medium was assayed for VEGF and progesterone. In study 1, LGC produced 8-fold greater levels of VEGF than NLGC (899 +/- 471 vs. 111 +/- 26 pg/mL, mean +/- SEM; P < 0.05). In vitro treatment with hCG increased (P < 0.05) VEGF production by NLGC to levels that were not different from the LGC incubated under control conditions. In vivo bolus doses of r-hCG (100 and 1000 IU) and r-hFSH (2500 IU) were equally effective in elevating granulosa cell VEGF production. In study 2, in vitro treatment with r hFSH, r-hLH, and hCG markedly increased (P < 0.05) VEGF and progesterone production by the NLGC in a dose- and time-dependent manner. By comparison, the three gonadotropins (100 ng/mL dose) only modestly increased VEGF and progesterone production by LGC. These experiments demonstrate a novel role for the midcycle surge of gonadotropin (LH/CG or FSH) in primates to promote VEGF production by granulosa cells in the periovulatory follicle. Further, the data demonstrate that FSH-like as well as LH-like gonadotropins directly stimulate VEGF synthesis by granulosa cells. PMID- 9215285 TI - Suppression of tyrosine kinase activity inhibits [3H]thymidine uptake in cultured human pituitary tumor cells. AB - Tyrosine kinases are involved in the phosphorylation of proteins that regulate cell growth and proliferation. The mitogenic effect of several growth factors requires tyrosine kinase activity of their receptors. The effect of inhibition of tyrosine kinase activity on thymidine uptake into cultured human pituitary adenoma cells was studied using two inhibitors, genestein and methyl-2,3 dihydroxycinnamate (MDHC). Of 33 pituitary adenomas, 7 incorporated sufficient [3H]thymidine to be investigated in the experiments. Genestein and MDHC both potently inhibited thymidine uptake into these tumors, with a mean inhibition by 74 mumol/L genestein of 61.96 +/- 18.96% (+/- SD inhibition of basal), by 740 mumol/L genestein of 92.65 +/- 8.59%, and by 100 mumol/L MDHC of 93.84 +/- 3.85%. The 7 pituitary adenomas were all large with suprasellar extension and secreted interleukin-6 in vitro. They included 2 prolactinomas, 1 somatotropinoma, 1 mammosomatropinoma, and 3 clinically nonfunctioning adenomas. Epidermal growth factor stimulated thymidine uptake in 2 of the 3 clinically nonfunctioning adenomas studied, and this stimulation was inhibited by genestein. Both of these tumors released FSH in cell culture and are probably silent gonadotropinomas. The growth stimulatory effect of conditioned medium from human pituitary cell culture on GH3 cells was inhibited by both genestein and MDHC. We conclude that tyrosine kinase activity is crucial for the integrity and growth of pituitary adenomas in culture. Growth factors released by pituitary adenomas potentially may maintain and promote tumor growth by stimulating tyrosine kinase activity. PMID- 9215286 TI - Effects of gender, ethnicity, body composition, and fat distribution on serum leptin concentrations in children. AB - The Ob protein leptin has been shown to be closely correlated with measures of body fat in humans and animals. Studies have suggested that there are both gender and ethnic differences in serum leptin concentrations, even after controlling for total and relative body fat and body mass index. We hypothesized that gender and ethnic differences in serum leptin concentrations are due to differences in both body composition and body fat distribution. We measured fasting serum leptin concentration, body composition (fat mass and fat-free mass by dual energy x-ray absorptiometry), and body fat distribution (intraabdominal and sc abdominal adipose tissue by computed tomography) in 74 prepubertal boys and girls (43 African-Americans and 31 Caucasians). Our results showed that gender differences in serum leptin concentrations could not be fully explained by differences in body mass index, total fat mass, or relative body composition. However, when serum leptin concentrations were adjusted for differences in relative body composition (fat mass and fat-free mass) and body fat distribution (sc and intraabdominal adipose tissue), gender no longer had an independent effect on the serum leptin concentration. Serum leptin concentrations were not influenced by ethnicity. Thus, when comparing group differences in serum leptin concentrations, it is necessary to adequately control for group differences in body composition and fat distribution. PMID- 9215287 TI - Augmented hepatic and skeletal thyromimetic effects of tiratricol in comparison with levothyroxine. AB - A thyroid hormone analog with organ-selective effects could have therapeutic application for disorders such as hyperlipidemia and osteoporosis. We performed a randomized clinical trial to determine the specific thyromimetic effects of tiratricol. Twenty-four athyreotic patients underwent detailed metabolic and physiological evaluation after a 2-month baseline period, taking TSH-suppressive doses of L-T4. They were then randomized to blinded treatment with either tiratricol (24 micrograms/kg twice daily) or L-T4 (1.9 micrograms/kg daily). The dose of hormone was increased until the TSH level was less than 0.1 mU/L, and the metabolic and physiological testing was repeated. Comparing the change from baseline to the study drug periods, when serum TSH levels were equivalently suppressed, there were no significant differences between the two groups in resting metabolic rate, weight, urea nitrogen excretion, or symptom score. Plasma total and low density lipoprotein cholesterol levels declined 13 +/- 4% and 23 +/ 6% in the tiratricol group compared with 2 +/- 2% and 5 +/- 3% in the L-T4 group (P = 0.015 and P = 0.0066, respectively). Serum sex hormone-binding globulin levels increased 55 +/- 13% with tiratricol compared with a 1.7 +/- 4% decline with L-T4 (P = 0.0006), indicating an augmented hepatic response to tiratricol. Skeletal metabolic activity was enhanced, with increased levels of serum osteocalcin and urinary excretion of calcium and pyridinium cross-links. Tiratricol and L-T4 had comparable effects on cardiovascular function. Tiratricol has distinct augmented hepatic and skeletal thyromimetic actions of potential therapeutic value. PMID- 9215288 TI - Comparison of immunocytochemical and molecular features with the phenotype in a case of incomplete male pseudohermaphroditism associated with a mutation of the luteinizing hormone receptor. AB - We report the case of an infant who presented at birth with a hypoplastic phallus associated with hypospadias. Low testosterone production, normal serum levels of steroid precursors, and increased LH in response to LH-releasing hormone supported a defect in Leydig cell differentiation or function. Conventional microscopic study of the testes showed fibroblastic cells in the interstitium. However immunocytochemical analysis using anti-LH receptor and anti-P450c17 antibodies demonstrated that about one third of these cells were Leydig cells or precursors of Leydig cells. No histological feature could distinguish the latter cells from fibroblasts. A homozygous substitution of cysteine 133 for arginine was found in the extracellular domain of the receptor. This is the first naturally occurring missense mutation found in the extracellular domain of the LH receptor. COS-7 cells transfected with the mutant receptor exhibited a marked impairment of hCG binding, whereas some cAMP production could be observed at high hCG concentrations. We propose that the partial impairment of LH receptor function, as reflected by the presence of Leydig cells, was responsible for the incomplete male pseudohermaphroditism observed in our patient. PMID- 9215289 TI - Changes in serum immunoreactive and bioactive growth hormone concentrations in boys with advancing puberty and in response to a 20-hour estradiol infusion. AB - Acceleration of linear growth during puberty is associated with increased GH secretion, although the relationship between growth and GH is complex. As GH exists as a family of isoforms, some of which may not be identified by immunoassay, there may be alterations in isoform secretion during pubertal maturation that result in increased growth. The changes in serum immunoreactive and bioactive GH concentrations across pubertal maturation were determined in 30 boys, aged 6.5-19.3 yr, with idiopathic short stature or constitutional delay of adolescence. Data were grouped as follows: 1) 6 prepubertal boys with bone age 7 yr or less; 2) 5 prepubertal boys with bone age of more than 7 yr, 3) 10 boys in early puberty; 4) 9 boys with mid- to late puberty. Blood was obtained every 20 min from 2000-0800 h. An equal aliquot of each serum sample was pooled for determination of GH by bio- and immunoassays. The mean serum immunoreactive GH concentration increased from 2.1 +/- 0.3, 1.8 +/- 0.3, and 2.9 +/- 0.5 micrograms/L in groups 1, 2, and 3, respectively, to a peak of 4.6 +/- 0.7 micrograms/L in group 4 (P < 0.05 vs. groups 1-3). The mean serum GH bioactivity was 48 +/- 13 micrograms/L in group 1 and declined to 39 +/- 8 and 31 +/- 3 micrograms/L in groups 2 and 3, increasing to a maximum of 64 +/- 15 micrograms/L in group 4 (P < 0.05 vs. group 3). The ratio of bioactive to immunoreactive GH suggests that the biopotencies of secreted isoforms do not increase during pubertal maturation. The role of E2 in increasing GH secretion was characterized in 8 additional early pubertal boys. Each boy received a saline infusion from 1000-0800 h, followed 1 week later by an infusion of E2 at 4.6 nmol/m2.h. Blood was obtained every 15 min from 2200-0800 h for GH and LH and every 60 min for E2 and testosterone. An equal aliquot of each overnight serum sample was pooled for insulin-like growth factor I (IGF-I) and GH by immuno- and bioassays. The mean serum LH concentration decreased from 5.0 +/- 0.9 to 2.3 +/- 0.6 IU/L (P < 0.01), and the E2 concentration increased from 22 +/- 4 to 81 +/- 26 pmol/L (P < 0.01) during saline and E2 infusions, respectively. Mean serum GH concentrations as measured by immunoassay were similar during both infusions (6.6 +/- 1.4 vs. 9.7 +/- 2.1 micrograms/L; saline vs. E2 infusion, respectively). In contrast, the mean serum GH concentration, as measured by bioassay, decreased from 48 +/- 10 micrograms/L during saline infusion to 16 +/- 3 micrograms/L during E2 infusion (P < 0.05). The mean serum IGF-I concentration also decreased significantly from 116 +/- 17 to 93 +/- 15 micrograms/L (saline vs. E2 infusion, respectively; P < 0.05). Thus, although mean overnight serum GH concentrations increase in late puberty, whether measured by immuno- or bioassay, an acute increase in E2 produces an acute decline in serum GH bioactivity and a lesser decline in the serum IGF-I concentration. These unexpected changes indicate that E2 may affect pubertal growth and GH secretion in a complex or biphasic manner depending on the context in which it is administered. PMID- 9215290 TI - Peak and trough growth hormone concentrations have different associations with the insulin-like growth factor axis, body composition, and metabolic parameters. AB - GH is secreted in a pulsatile fashion, promoting growth and anabolism. The components of the pulsatile signal involved in these diverse effects are unclear. We constructed (20-min sampling interval) and analyzed 24-h serum GH profiles in 45 adult male volunteers, 59.4-69.9 yr old, body mass index (BMI) 21.9-36.5 Kg/m2, using Fourier transformation and a concentration distribution analysis that determines the concentration at or below which the serum GH concentrations in the 24-h profile spend a percentage of the total time. The observed concentrations (OC) below which 95% and 5% of the values in the time series lie [lsb]OC95 (peaks) and OC5 (troughs)] and mean 24-h serum GH concentrations were related to measures of the insulin-like growth factor (IGF) family, parameters of body composition, fasting insulin and cholesterol measures, and GH-binding protein concentrations. Mean 24-h serum GH concentrations ranged between 0.19 and 2.15 mU/L (1 microgram/L = 2.6 mU/L). Pulse periodicity was between 180 and 200 min. There was a positive relationship between peak GH levels and serum IGF-1 and IGFBP-3 levels (r = 0.39; P = 0.009 and r = 0.32; P = 0.03, respectively). GH trough levels were unrelated to these measures of the IGF family. In contrast, GH troughs were related inversely to BMI (r = -0.31; P = 0.04) and waist-hip ratio (r = -0.4; P = 0.006). Peak GH levels were not related to these measures. Factors known to influence these measures, fasting insulin concentration, or cortisol secretion did not alter the trough GH relationship in multiple regression analysis. All GH parameters were related inversely to fasting insulin concentration. Although GH parameters were related inversely to cholesterol and low-density lipoprotein-cholesterol, this effect disappeared when age and fasting insulin levels were introduced into the regression. GH-binding protein levels related most strongly to BMI (r = 0.60; P < 0.001), with no effect of any GH parameter observed in multiple regression analysis. These results suggest that the peak values of a GH concentration profile may influence the IGF axis, whereas trough values may influence body composition and metabolic parameters of GH action. PMID- 9215291 TI - Free insulin-like growth factor I (IGF-I) in healthy subjects: relationship with IGF-binding proteins and insulin sensitivity. AB - The majority of insulin-like growth factor I (IGF-I) circulates in blood bound to a family of IGF-binding proteins (IGFBPs). Only a small fraction of IGF-I is unbound or free, and one of the postulated roles of the IGFBPs is regulation of this free component, thereby increasing IGF-I bioavailability. Whether free IGF-I plays a physiological role in glucose homeostasis, however, is not clear. In this study, we examined the effects of acute changes in serum insulin on free IGF-I, total IGF-I, IGFBP-1, and IGFBP-3 in 11 healthy subjects. Glucose (0.3 g/kg) and insulin (0.05 U/kg) were injected iv at 0 and 20 min, respectively. Blood samples were drawn at defined intervals for 3 h, and insulin sensitivity (SI) was computed by Bergman's minimal model. Serum insulin reached a first peak after glucose injection and a second, higher peak after exogenous insulin administration. Although the total IGF-I level remained constant for the duration of the experiment, free IGF-I decreased by 20% 20 min after the first insulin peak and by 35% 20 min after the second peak. IGFBP-1 first declined to 20% below basal, then rose to 3-fold the basal level. IGFBP-3 increased linearly to 20% above basal by the end of the experiment, and this increase mirrored the decline of free IGF-I. In the fasting state, free IGF-I was positively correlated with SI (r = 0.52; P < 0.005) and inversely correlated with glucose (r = -0.51; P < 0.005) and IGFBP-1 (r = -0.65; P < 0.001). In conclusion, free IGF-I is acutely regulated by insulin and correlates with SI, suggesting that it may play a physiological role in glucose homeostasis. PMID- 9215292 TI - Exercise and circadian rhythm-induced variations in plasma cortisol differentially regulate interleukin-1 beta (IL-1 beta), IL-6, and tumor necrosis factor-alpha (TNF alpha) production in humans: high sensitivity of TNF alpha and resistance of IL-6. AB - Although we have previously shown that the integrity of inflammatory mediator induced activation of the hypothalamic-pituitary-adrenal axis is essential for conferring resistance to inflammatory disease in susceptible Lewis rats, the role of endogenous glucocorticoid secretion in human immune function in either health or disease is less clear. To further understand the relevance of physiological variations in plasma cortisol on immune function in humans, we evaluated ex vivo lipopolysaccharide-induced interleukin-1 beta (IL-1 beta), IL-6, and tumor necrosis factor-alpha (TNF alpha) production in the whole blood of healthy volunteers studied under conditions chosen to approximate either physiological or pharmacological glucocorticoid levels. Administration of a pharmacological dose of hydrocortisone suppressed the production of all three cytokines, whereas administration of a physiological dose of hydrocortisone suppressed only TNF alpha production. Stress-induced levels of glucocorticoids, achieved during exercise at 100% maximal oxygen utilization, suppressed IL-1 beta and TNF alpha production, but were without effect on IL-6 production. In addition, circadian variations of cortisol were associated with decreased TNF alpha production, but were without effect on IL-1 beta or IL-6 production. These studies challenge the generally accepted idea that glucocorticoids consistently suppress cytokine production and indicate a hierarchy of sensitivity, with TNF alpha having the greatest sensitivity, IL-1 beta having intermediate sensitivity, and IL-6 being resistant. The resistance of IL-6 production to glucocorticoid suppression is compatible with data suggesting an antiinflammatory as well as a proinflammatory action for this cytokine. PMID- 9215293 TI - Epidermal growth factor and sex steroids dynamically regulate a marker of endometrial receptivity in Ishikawa cells. AB - The factors regulating human endometrial receptivity remain poorly understood. The alpha v beta 3 integrin cell adhesion molecule appears to be regulated in the human endometrium, appearing on postovulatory days 5-6, corresponding to the time of initial embryo attachment. This integrin has been extensively studied as a potential marker of endometrial receptivity and is aberrantly expressed in the endometrial epithelium of some infertile women. Ishikawa cells are a well differentiated endometrial adenocarcinoma cell line that maintain functional estrogen and progesterone receptors and are a useful model to study steroid mediated events in human endometrial epithelium. This cell line expresses most of the normal endometrial epithelial integrins, including the alpha v beta 3 vitronectin receptor. The regulation of this integrin was studied with fluorescence immunocytochemistry, flow cytometry, and Northern blot analysis. Estrogen with or without progesterone treatment down-regulates alpha v beta 3 in this cell line. Several growth factors, including epidermal growth factor and the closely related transforming growth factor-alpha significantly increase the expression of this integrin. We conclude that endometrial differentiation is influenced by both steroid hormones and growth factors. The alpha v beta 3 integrin appears to be an excellent marker to study the molecular events leading to the establishment of uterine receptivity and successful implantation. PMID- 9215294 TI - Decreased expression of Wilms' tumor gene WT-1 and elevated expression of insulin growth factor-II (IGF-II) and type 1 IGF receptor genes in prostatic stromal cells from patients with benign prostatic hyperplasia. AB - Benign prostatic hyperplasia (BPH) is a common proliferative disorder of unknown etiology. We have previously documented that the insulin-like growth factor (IGF) axis is critical for prostate cell growth and is abnormal in BPH. The type 1 IGF receptor (IGF-1R) is constitutively expressed by most body tissues and plays a significant role in regulating cell proliferation, consistent with the role of its ligands (IGF-I and IGF-II) as important mitogenic factors. The Wilms' tumor gene product (WT-1) is a tumor suppressor that has been shown to be altered in rare kidney tumors and is known to regulate IGF-II and IGF-1R. We investigated the possibility that the expression of prostatic WT-1, IGF-1R, and IGF-II genes is altered in patients with BPH. We utilized primary cultures of prostatic stromal cells grown from normal (n = 9) and hyperplastic (n = 9) surgical specimens and analyzed WT-1, IGF-1R, and IGF-II messenger RNA levels. In all of the BPH cell strains, WT-1 expression (measured by RT-PCR and RNase protection assays) was strikingly lower than that found in normal strains (0-20% of normal, mean 14% of normal, P < 0.01). The expression of both the IGF-1R (300% of normal, P < 0.05) and IGF-II (1000% of normal, P < 0.01) messenger RNAs was higher in BPH strains as compared with normal strains. No changes were seen in stromal cell strains derived from prostatic adenocarcinoma. Thus, in cultured human prostatic stromal cell strains from patients with BPH, decreased WT-1 gene expression is associated with increases in the expression of the IGF-1R and IGF-II genes that are known transcriptional targets of WT-1. These findings indicate that reduced expression of the WT-1 tumor suppressor gene and elevated IGF-1R and IGF-II gene expression may be involved in the pathophysiology of prostatic hyperplasia, implying a new role for the Wilms' tumor gene in nonmalignant states. PMID- 9215296 TI - Differential gene expression of steroid 5 alpha-reductase 2 in core needle biopsies from malignant and benign prostatic tissue. AB - Androgens are implicated in the development of prostate cancer (CAP) and benign prostate hyperplasia. The conversion of testosterone to the more potent metabolite dihydrotestosterone by prostatespecific steroid 5 alpha-reductase type 2 (5 alpha-red2) is a key mechanism in the action of androgens in the prostate and is important in the promotion and progression of prostate diseases. Manipulation of the turnover of androgens is thus fundamental in the pharmacological treatment strategy. We have developed a sensitive solution hybridization method for quantification of the gene expression of 5 alpha-red2 in core needle biopsies of the prostate. The 5 alpha-red2-specific messenger RNA (mRNA) levels were measured in 50 human prostate transrectal ultrasound-guided core biopsies obtained from 31 outpatients (median age 72, range 67-88 yr) undergoing biopsy for diagnostic purposes. Significant differences were observed in the gene expression of 5 alpha-red2 between cancerous and noncancerous tissue. In the 14 biopsies judged cancerous, the median 5 alpha-red mRNA levels were 3.5 amol/ng total RNA compared with 12.0 amol/ng total RNA in the biopsies showing no cancer (P = 0.0018). The median 5 alpha-red2 mRNA level in noncancerous tissue was thus 3.4 times higher than in the cancerous specimens. PMID- 9215295 TI - Serum levels of insulin-like growth factor-I (IGF-I) and IGF-binding protein-3 in healthy centenarians: relationship with plasma leptin and lipid concentrations, insulin action, and cognitive function. AB - It has been demonstrated that healthy centenarians have more favorable anthropometric characteristics and insulin-mediated glucose uptake than aged subjects. The plasma insulin-like-growth factor I (IGF-I) concentration may account for such differences. Three groups of subjects were studied: 1) adults (< 50 yr; n = 30), 2) aged subjects (75-99 yr; n = 30), 3) centenarians (> 100 yr; n = 19). In all subjects, fasting plasma IGF-I, IGF-binding protein-3 (IGFBP-3), leptin, and lipid concentrations were determined; body composition was assessed by bioimpedance analysis; and insulin-mediated glucose up-take was evaluated by euglycemic hyperinsulinemic glucose clamp. IGF-I declined with advancing age, but no differences between aged subjects and centenarians were found. IGFBP-3 showed a trend similar to IGF-I, but lower values were present in centenarians than in aged subjects. Nevertheless, centenarians had a plasma IGF-I/IGFBP-3 molar ratio greater than that in aged subjects. Centenarians had also a whole body glucose disposal (WBGD) greater than that in aged subjects, but similar to that in adults. Mini Mental State Examination (27 +/- 2.1 vs. 18.3 +/- 3.1; P < 0.02) and Instrumental Activities Daily Living (26 +/- 2.6 vs. 8.4 +/- 4.1; P < 0.001) scores were significantly different in aged subjects and centenarians, respectively. In centenarians, the plasma IGF-I/IGFBP-3 molar ratio correlated with the body mass index (r = -0.55; P < 0.009); the amount of body fat (r = 0.62; P < 0.003); fat-free mass (r = 0.56; P < 0.008); fasting plasma leptin (r = -0.63; P < 0.004), triglycerides (r = -0.58; P < 0.01), free fatty acid (r = 0.64; P < 0.005), and low density lipoprotein cholesterol (r = -0.59; P < 0.009) concentrations; Mini Mental State Examination (r = 0.53; P < 0.0.03); and WBGD (r = 0.64; P < 0.005). All correlations were independent of daily fat and carbohydrate intake and WBGD (P < 0.05 for all). No significant correlations between the plasma IGF-I/IGFBP-3 molar ratio and plasma total (r = 0.31; P = NS) and high density lipoprotein cholesterol (r = 0.34; P = NS) concentrations were present. The correlation between the plasma IGF-I/IGFBP-3 molar ratio and WBGD persisted after adjustment for body fat, fasting plasma insulin concentration, daily carbohydrate and fat intake, and daily physical activity (r = 0.55; P < 0.009), but not after further adjustment for plasma free fatty acid concentration (r = 0.30; P = 0.17). In conclusion, healthy centenarians have plasma IGF-I/IGFBP 3 molar ratio greater than aged subjects. A more elevated plasma IGF-I/IGFBP-3 molar ratio might improve insulin action and plasma lipid concentration in centenarians. PMID- 9215297 TI - Preservation of growth hormone pulsatility despite pituitary pathology, surgery, and irradiation. AB - Detailed assessment of physiological and pathophysiological GH secretion has, until recently, been limited by the poor sensitivity of the available assays. We have used an ultrasensitive chemiluminescence GH assay (sensitivity, 0.002 microgram/L) to study 24-h GH profiles (20-min sampling) from 24 patients who had been treated for hypothalamic-pituitary disease with surgery and irradiation and from 24 healthy control subjects matched for age, sex, and body mass index. Twenty-three of the 24 patients demonstrated pulsatile GH secretion, determined by Cluster. The median (range) area under the curve for GH, mean pulse area, mean pulse height, average valley mean level, and mean interpeak nadir were lower in the patients than in the controls [119.25 (7.273-843.600) vs. 968.539 (227.200 4625.000) min/microgram.L (P < 0.00001); 3.777 (0.288-30.850) vs. 61.390 (12.880 224.210) min/microgram.L (P < 0.00001), 0.107 (0.010-0.958) vs. 1.408 (0.368 5.050) micrograms/L (P < 0.00001), 0.074 (0.006-0.415) vs. 0.348 (0.048-2.350) microgram/L (P < 0.00001), and 0.066 (0.003-0.270) vs. 0.205 (0.021-1.838) microgram/L (P = 0.0004), respectively]. The median (range) number of pulses, mean pulse duration, and mean interval between pulses did not differ between the patients and controls [10 (4-15) vs. 10 (7-15; P = 0.36), 96.4 (68.0-220.0) vs. 104.0 (72.0-151.4) min (P = 0.65) and 128.0 (92.8-255.0) vs. 126.2 (90.0-180.0) min (P = 0.73), respectively]. The diurnal rhythm of GH secretion was present in the controls, but there was only limited evidence of residual diurnal rhythm in the patients. This study has demonstrated that GH secretion remains pulsatile in GH-deficient patients despite the mass effect of hypothalamic-pituitary pathology, pituitary surgery, and radiotherapy. With the development of potent GH secretagogues that are active orally, our findings may have important implications for the future management of GH-deficient subjects. PMID- 9215298 TI - gamma-Interferon-induced resistance to 1,25-(OH)2 D3 in human monocytes and macrophages: a mechanism for the hypercalcemia of various granulomatoses. AB - The hypercalcemia of various granulomatoses is caused by endogenous 1,25 dihydroxyvitamin D [1,25-(OH)2D3] overproduction by disease-activated macrophages. The inability of 1,25(OH)2D3 to suppress its synthesis in macrophages contrasts with the tight control of its production in macrophage precursors, peripheral blood monocytes (PBM). We examined whether 1,25(OH)2D3 resistance develops as PBM differentiate to macrophages or with macrophage activation. Normal human pulmonary alveolar macrophages (PAM) are less sensitive to 1,25(OH)2D3 than PBM, despite similar vitamin D receptor content; however, both PBM and PAM respond to exogenous 1,25-(OH)2D3 by inhibiting 1,25(OH)2D3 synthesis and inducing 1,25(OH)2D3 degradation through enhancement of 24 hydroxylase mRNA levels and activity. The human monocytic cell line THP-1 mimics PAM in 1,25(OH)2D3 synthesis and sensitivity to exogenous 1,25(OH)2D3. We utilized THP-1 cells to examine the response to 1,25(OH)2D3 with macrophage activation. Activation of THP-1 cells with gamma-interferon (gamma-IFN) enhances 1,25(OH)2D3 synthesis 30-fold, blocks 1,25-(OH)2D3 suppression of its synthesis, and reduces by 42.2% 1,25-(OH)2D3 induction of its degradation. The antagonistic effects of gamma-IFN are not merely restricted to enzymatic activities. In THP-1 cells and in normal PBM, gamma-IFN inhibits 1,25-(OH)2D3 induction of 24 hydroxylase mRNA levels without reducing mRNA stability, suggesting gamma-IFN inhibition of 1,25(OH)2D3 transactivating function. These results explain 1,25(OH)2D3 overproduction in granulomatoses and demonstrate potent inhibition by gamma-IFN of 1,25(OH)2D3 action in immune cells. PMID- 9215299 TI - Tonic support of luteinizing hormone secretion by adrenal progesterone in the ovariectomized monkey replaced with midfollicular phase levels of estradiol. AB - Although it is known that progesterone facilitates the estradiol-induced gonadotropin surge at midcycle, its effect on LH secretion at other times of the follicular phase remains to be investigated. In this study, we investigate the role of progesterone on tonic LH secretion in the ovariectomized primate replaced with estradiol at levels representative of the follicular phase. The experiments were performed in nine ovariectomized rhesus monkeys, either unreplaced with estradiol or after a 5-day estradiol therapy to mimic early follicular (10-36 pg/mL; low dose) and midfollicular (medium dose; 40-75 pg/mL) concentrations. We used two antiprogesterone compounds, RU-486 (5 mg) and ORG-31806 (1 mg), to antagonize endogenous progesterone activity and studied their acute effects on LH secretion in each group. LH concentrations were measured at 15-min intervals for a 3-h baseline period and during a 5-h period after antagonist administration. LH concentrations remained unchanged after either antiprogesterone compound or diluent (ethanol) administration in the estrogen-unreplaced monkeys or after low dose estradiol replacement. However, both antiprogesterone compounds significantly decreased LH secretion in monkeys pretreated with the medium dose of estradiol; by 5 h, the mean (+/-SE) areas under the LH curve were 54.8 +/- 4.1% and 64.0 +/- 4.2% after RU-486 and ORG-31806, respectively (P < 0.05 vs. unreplaced and low dose estrogen-replaced groups). To exclude the possibility that the LH response reflects an agonist action of the progesterone antagonist, LH responses to progesterone infusions (at three doses to reproduce preovulatory, luteal, and pharmacological levels) were also examined in monkeys pretreated with midfollicular levels of estradiol. In none of these was there a decrease in LH; rather, progesterone infusions resulted in an increase in LH secretion in all three groups (to 115-194% of baseline in seven of eight monkeys). Finally, we determined that at the dose used in our protocol, neither of the two progesterone antagonists was able to prevent dexamethasone-induced cortisol suppression, thus excluding the possibility that results after progesterone antagonist administration may reflect a putative antiglucocorticoid activity of these compounds. When the doses of the antiprogesterone compounds were increased 6 times, only RU-486 counteracted the effect of dexamethasone on cortisol. In summary, our data indicate support by progesterone of tonic LH secretion in the nonhuman primate under estrogenic conditions similar to the midfollicular phase of the menstrual cycle. Significantly, because the experiments were performed in ovariectomized monkeys, and endogenous progesterone was most probably of adrenal origin, the data also demonstrate a role of the hypothalamo-pituitary-adrenal axis in support of gonadotropin secretion. PMID- 9215300 TI - Evidence for an inhibitory effect of physiological levels of insulin on the growth hormone (GH) response to GH-releasing hormone in healthy subjects. AB - It has been previously reported that in healthy subjects, the acute reduction of free fatty acids (FFA) levels by acipimox enhances the GH response to GHRH. In the present study, the GH response to GHRH was evaluated during acute blockade of lipolysis obtained either by acipimox or by insulin at different infusion rates. Six healthy subjects (four men and two women, 25.8 +/- 1.9 yrs old, mean +/- SE) underwent three GHRH tests (50 micrograms iv, at 1300 h) during: 1) iv 0.9% NaCl infusion (1200-1500 h) after oral acipimox administration (250 mg) at 0700 h and at 1100 h; 2) 0.1 mU.kg-1.min-1 euglycemic insulin clamp (1200-1500 h) after oral acipimox administration (250 mg at 0700 h and at 1100 h); 3) 0.4 mU.kg-1.min-1 euglycemic insulin clamp (1200-1500 h) after oral placebo administration (at 0700 and 1100 h). Serum insulin (immunoreactive insulin) levels were significantly different in the three tests (12 +/- 2, 100 +/- 10, 194 +/- 19 pmol/L, P < 0.06), plasma FFA were low and similar (0.04 +/- 0.003, 0.02 +/- 0.005, 0.02 +/- 0.003, not significant), and the GH response to GHRH was progressively lower (4871 +/- 1286, 2414 +/- 626, 1076 +/- 207 micrograms/L 120 min), although only test 3 was significantly different from test 1 (P < 0.05). Pooling the three tests together, a significant negative regression was observed between mean serum immunoreactive insulin levels and the GH response to GHRH (r = -0.629, P < 0.01). Our results indicate that in healthy subjects, acipimox and hyperinsulinemia produce a similar decrease in FFA levels and that at similar low FFA, the GH response to GHRH is lower during insulin infusion than after acipimox. These data suggest that insulin exerts a negative effect on GH release. Because the insulin levels able to reduce the GH response to GHRH are commonly observed during the day, for instance during the postprandial period, we conclude that the insulin negative effect on GH release may have physiological relevance. PMID- 9215301 TI - Soluble interleukin-1 receptor antagonist serum levels in smokers and nonsmokers with Graves' ophthalmopathy undergoing orbital radiotherapy. AB - Interleukin-1 (IL-1) plays an important role in the pathogenesis of Graves' ophthalmopathy (GO). Impaired antagonism of the proinflammatory cytokine IL-1 by the naturally occurring IL-1 receptor antagonist (IL-1RA) has been implicated in the initiation and perpetuation of various autoimmune diseases and may play a role in the evolution of GO. Cigarette smoking appears to adversely affect the course of GO. We have evaluated the course of IL-1 alpha, IL-1 beta, and soluble IL-1RA (sIL-1RA) serum levels in smokers and nonsmokers with GO undergoing orbital radiotherapy (OR). We prospectively studied the eye status of 27 randomly selected patients (mean age 47.3 +/- 11.0 yr; 20 females; 18 smokers) with active, moderately severe GO before and 3 and 6 months following OR, respectively. None had received any previous treatment for GO, and all patients were kept euthyroid on carbimazole. Serum concentrations of IL-1 alpha, IL-1 beta, and sIL-1RA were measured using highly sensitive enzyme linked immunosorbent assay systems. Baseline sIL-1RA levels were negatively correlated with the number of cigarettes smoked before and following OR (P < 0.0001). Patients with no or minor therapeutic response to OR (n = 8), all of whom were smokers, revealed mean baseline sIL-1RA levels of 114 +/- 85 pg/mL, which increased to 172 +/- 103 pg/mL at 3 months and 149 +/- 96 pg/mL at 6 months after initiation of OR, respectively. By contrast, patients with a good clinical response (n = 19, 9 nonsmokers), revealed significantly higher baseline sIL-1RA levels at 294 +/- 148 pg/mL (P = 0.004), which increased to 845 +/- 668 pg/mL at 3 months (P = 0.01) and 634 +/- 337 pg/mL at 6 months (P < 0.001), respectively, following initiation of OR. Serum concentrations of IL-1 alpha IL-1 beta were below 3.9 pg/mL in all patients with GO who were studied, and were not correlated with gender, age, smoking status, clinical course, or outcome. Low baseline levels and impaired surge of sIL-1RA serum levels following OR were strongly correlated with smoking status and a less favorable therapeutic outcome in patients with active, moderately severe GO. Measurement of sIL-1RA may contribute to predict the therapeutic response to OR in patients with active, moderately severe GO. Strategies designed to raise local or systemic concentrations of sIL 1RA may be of benefit to patients with GO. PMID- 9215303 TI - Paraoxonase polymorphism (Gln192-Arg) is associated with coronary heart disease in Japanese noninsulin-dependent diabetes mellitus. AB - Serum paraoxonase/arylesterase (PONA) is associated with high-density lipoprotein and may prevent oxidation of low-density lipoprotein by hydrolyzing lipid peroxides. A recent report suggested an association of glutamine (A type)/arginine (B type) polymorphism at position 192 of PONA gene with coronary heart disease (CHD) among Caucasian patients with noninsulin-dependent diabetes mellitus (NIDDM). However, conflicting results have also been reported. To investigate the significance of this polymorphism in the pathogenesis of CHD, we performed an association study of this polymorphism with CHD in Japanese NIDDM patients. We genotyped 164 patients with NIDDM, 42 with CHD, and 122 without CHD. Other known risk factors for CHD were matched between the 2 groups. AB+BB isoforms were detected in 41 of 42 diabetic patients with CHD. The proportion of B allele carriers (AB+BB) was significantly higher than that of AA carriers among diabetic patients with CHD compared with those without CHD (chi 2 = 7.68, P = 0.003). Multivariate logistic regression analyses showed a markedly increased odds ratio (OR: 8.823, CI, 1.13-68.7) in B allele carriers, while ORs of other risk factors remained between 1.01 and 1.92. Carriers of the B allele of the Gln192Arg polymorphism in the PONA gene proved to be at increased risk for developing CHD in Japanese NIDDM patients. This association was independent of other known risk factors for CHD, suggesting an important role of the paraoxonase B isoform in the pathogenesis of CHD. PMID- 9215302 TI - Determinants of abnormal gonadotropin secretion in clinically defined women with polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a heterogeneous disorder of reproductive age women characterized in its broadest definition by the presence of oligoamenorrhea and hyperandrogenism and the absence of other disorders. Defects of gonadotropin secretion, including an elevated LH level, elevated LH to FSH ratio, and an increased frequency and amplitude of LH pulsations have been described, but the prevalence of these defects in a large, unbiased population of PCOS patients has not been determined. Sixty-one women with PCOS defined by oligomenorrhea and hyperandrogenism and 24 normal women in the early follicular phase had LH samples obtained every 10 min for 8-12 h. Pool LH levels from the frequent sampling studies were within the normal range in the 9 PCOS patients (14.8%) who were studied within 21 days after a documented spontaneous ovulation. Excluding these post-ovulatory patients, 75.0% of the PCOS patients had an elevated pool LH level (above the 95th percentile of the normal controls), and 94% had an elevated LH to FSH ratio. In the anovulatory PCOS patients, pool LH correlated positively with 17-OH progesterone (R = 0.30, P = 0.03), but not with estradiol, estrone, testosterone, androstenedione, or DHEA-S. Pool LH and LH to FSH ratio correlated positively with LH pulse frequency (R = 0.40, P = 0.004 for pool LH, and R = 0.39; P = 0.005 for LH/FSH). There was also a strong negative correlation between pool LH and body mass index (BMI) (R = -0.59, P < 10(-5)). The relationship between BMI and LH secretion in the PCOS patients appeared to be strongest with body fatness, as pool LH was correlated inversely with percent body fat, whether measured by skinfolds (R = -0.61, P < 10(-5)), bioimpedance (R = -0.55, P < 10( 4)), or dual energy x-ray absorptiometry (DEXA) (R = -0.70, P = 0.001; n = 18 for DEXA only). By DEXA, the only body region that was highly correlated with pool LH was the trunk (R = -0.71, P = 0.001). The relationship between body fatness and LH secretion occurred via a decrease in LH pulse amplitude (R = -0.63, P < 10(-5) for BMI; R = -0.58, P < 10(-4) for bioimpedance; and R = -0.64, P = 0.004 for whole body DEXA), with no significant change in pulse frequency with increasing obesity (R = -0.17, P = 0.23 for BMI). IN CONCLUSION: 1) the prevalence of gonadotropin abnormalities is very high in women with PCOS selected on purely clinical grounds, but is modified by recent spontaneous ovulation; 2) the positive relationship between LH pulse frequency and both pool LH and LH to FSH ratio supports the hypothesis that a rapid frequency of GnRH secretion may play a key etiologic role in the gonadotropin defect in PCOS patients; 3) pool LH and LH pulse amplitude are inversely related to body mass index and percent body fat in a continuous fashion; and 4) the occurrence of a continuous spectrum of gonadotropin abnormalities varying with body fat suggests that nonobese and obese patients with PCOS do not represent distinct pathophysiologic subsets of this disorder. PMID- 9215304 TI - In obesity, glucose load loses its early inhibitory, but maintains its late stimulatory, effect on somatotrope secretion. AB - Glucose load has a biphasic effect on GH secretion. In fact, in normal subjects, glucose load has a prompt inhibitory and a late stimulatory effect on both spontaneous and GHRH-induced GH levels. The mechanism underlying the inhibitory effect is probably mediated by the increase in hypothalamic somatostatin, whereas that underlying the stimulatory effect is unclear. On the other hand, in obesity, a reduced somatotrope responsiveness to all GH secretagogues is well known, whereas recently, we found that glucose load, but not pirenzepine and somatostatin, fails to inhibit the GHRH-induced GH rise. Thus, the inhibitory effect of hyperglycemia on GH secretion is selectively lacking in obesity. The aim of the present study was to verify whether in obesity the late stimulatory effect of glucose on GH secretion is preserved. We studied 15 female obese patients (OB; age, 33.9 +/- 2.6 yr; body mass index, 36.4 +/- 1.5 kg/m2; waist/hip ratio, 0.9 +/- 0.1) and 12 normal female subjects (NS; 26.5 +/- 1.0 yr; 21.4 +/- 0.3 kg/m2) as controls. Two studies were performed. In study A (six OB and six NS) we evaluated the somatotrope response to GHRH (1 microgram/kg, i.v., at 0 min) alone or preceded by oral glucose (OGTT; 100 g, orally, at -45 min). In study B (nine OB and six NS) we studied the somatotrope response to OGTT (100 g, orally, at 0 min), saline plus GHRH (1 microgram/kg, iv, at 150 min), and OGTT plus GHRH. In study A, the GHRH-induced GH rise in NS was higher (P < 0.01) than that in OB. OGTT blunted the GHRH-induced GH rise in NS (0-90 min area under the curve, 318.9 +/- 39.1 vs. 696.3 +/- 110.8 micrograms/min-L; P < 0.05), but failed to modify it in OB (289.1 +/- 51.7 vs. 283.9 +/- 44.0 micrograms/min-L). In study B, the GHRH-induced GH rise in NS was higher (P < 0.01) than that in OB. OGTT induced a late GH increase in both NS (150-240 min area under the curve, 249.6 +/ 45.2 micrograms/min-L) and OB (103.2 +/- 31.4 micrograms/min-L). Moreover, OGTT enhanced the GHRH-induced GH rise in NS as well as in OB [1433.0 +/- 202.0 vs. 967.9 +/- 116.3 micrograms/min-L (P < 0.03) and 763.8 +/- 131.0 vs. 278.1 +/- 52.3 micrograms/min-L (P < 0.01), respectively]. The GH responses to OGTT alone and combined with GHRH in OB were lower (P < 0.03) than those in NS. Our data show that in human obesity, the oral glucose load loses its precocious inhibitory effect on the GHRH-induced GH rise but maintains its late stimulatory effect on somatotrope secretion. These findings suggest that the inhibitory and stimulatory effects of glucose load on GH secretion are unlikely to be due to biphasic modulation of hypothalamic somatostatin release, which seems selectively refractory to stimulation by hyperglycemia in obesity. PMID- 9215305 TI - Kinetics of insulin-like growth factor (IGF) and IGF-binding protein responses to a single dose of growth hormone. AB - The in vivo physiological relationships among GH, the insulin-like growth factors (IGFs), and the IGF-binding proteins (IGFBPs) are not completely defined, and single random measurements of these serum proteins do not completely reveal their dynamic relationships. We report the kinetic responses of the IGFs and IGF binding proteins to exogenous GH in 23 subjects with untreated GH deficiency [5 women and 18 men; age, 15.0 +/- 6.2 yr (+/- s.d.), height z-score = -4.4 +/- 2.2 (+/- s.d.); body mass index = 19.3 +/- 2.4 kg/m2]. After an overnight fast, subjects were given a sc dose of recombinant human GH (2.85 i.u./m2), and blood was sampled from an indwelling peripheral venous catheter 0, 1, 2, 3, 4, 5, 6, 8, 10, 12, 14, and 24 h after the injection. Subjects were then treated with recombinant human GH (2.85 i.u./m2.day); fasting samples were obtained at 3 months (n = 22), and timed sampling was repeated at 6 months (n = 21). Fasting levels of IGF-I, free IGF-I, IGF-II, IGFBP-3, and insulin increased significantly within 3 months of GH treatment, whereas IGFBP-1, IGFBP-2, and IGFBP-6 showed no change. In the timed sampling studies at 0 and 6 months, GH levels peaked 3 h after treatment; the degree of rise and the rate of decline were both greater at 6 months. IGF-I levels increased beginning at 4 h, continuing throughout the 24-h period at month 0, whereas a plateau was observed after 6-8 h during the 6-month study. Free IGF-I paralleled total IGF-I except during fasting, when it varied inversely with IGFBP-1. IGFBP-3 and IGF-II both showed late (> 20 h) responses to a dose of GH, whereas IGFBP-2 and IGFBP-6 showed minimal changes. IGFBP-1 varied inversely with insulin, which, in turn, varied with meal intake. Comparative studies in 2 subjects with GH receptor deficiency showed no response to exogenous GH. However, both IGFBP-1 and IGFBP-2 were several-fold elevated, and IGFBP-1 varied inversely with the low insulin levels. Our data are the first to examine multiple elements of the serum IGF system in response to GH in both GH-deficient and replete states. The relationships of the different response patterns provide insight into the physiology of this system and may guide future studies. PMID- 9215306 TI - Leptin production during moderate-intensity aerobic exercise. AB - Leptin, the protein product of the ob gene, may be involved in the regulation of energy balance. Although a clear relationship between energy intake and plasma leptin concentrations has been demonstrated in humans, little is known about the effect of exercise on leptin metabolism. In the present study, we evaluated abdominal adipose tissue leptin production in vivo by arteriovenous balance at rest and during 60 min of moderate-intensity cycle ergometer exercise (50% of maximal heart rate) in five sedentary male subjects (mean age 38.4 +/- 1.7 yr, body mass index (28.4 +/- 4.2 kg/m2). Blood samples were taken simultaneously from an abdominal vein, draining sc adipose tissue, and a radial artery, at rest and every 10 min during exercise. Adipose tissue blood flow was determined by the xenon washout technique. Plasma leptin concentrations did not change throughout exercise and were the same as the values obtained during resting conditions. Average net adipose tissue leptin production rates during exercise (3.07 +/- 0.89 ng/100 g-1.min-1) also were similar to resting values (3.86 +/- 0.95 ng/100 g 1.min-1). These results demonstrate that plasma leptin concentrations and leptin production do not change during an acute bout of moderate-intensity aerobic exercise. PMID- 9215307 TI - Allelic loss in parathyroid tumors from individuals homozygous for multiple endocrine neoplasia type 1. AB - Homozygosity for the multiple endocrine neoplasia type 1 (MEN1) gene mutation was described in two of three affected siblings of a kindred in which both parents and the third daughter were heterozygotes. Surprisingly, in the two homozygotes, the disease history did not differ from the one of the heterozygotes. In the attempt to unravel genetic differences in parathyroid tumorigenesis between homozygotes and heterozygotes, restriction fragment length polymorphism analysis and microsatellite PCR analysis for loss of heterozygosity (LOH) at the MEN1 gene region on chromosome 11q13 was performed in parathyroid tissues removed at surgery from the mother, her heterozygous sister, and the three siblings. Allelic losses were evidenced in the larger glands of each patient, with a similar pattern of chromosome 11q12-13 losses. The somatic mutation consisted of a large lose of genetic material from chromosome 11. No gross differences exist in the 11q12-13 LOH observed between homozygous and heterozygous carriers. Interestingly, one of the parathyroid tumors from one heterozygote exhibited region of skipped LOH at the 11q12-13 region. The region in the depth of the critical interval retained heterozygosity, whereas those flanking it shared LOH. These findings indicate that inactivation of both copies of the MEN1 gene are not sufficient for parathyroid tumor development in MEN 1 patients and that tumor suppressor genes, other than the MEN1 gene on chromosome 11 or on other chromosomes, can be involved in the pathogenesis of parathyroid tumorigenesis in MEN 1 syndrome. PMID- 9215308 TI - Hyperinsulinemia and decreased insulin-like growth factor-binding protein-1 are common features in prepubertal and pubertal girls with a history of premature pubarche. AB - The fasting insulin resistance index, mean blood glucose, mean serum insulin (MSI), early insulin response to glucose, glucose uptake rate in peripheral tissues, and insulin sensitivity indexes in response to a standard oral glucose tolerance test; serum insulin-like growth factor I (IGF-I), IGF-binding protein-1 (IGFBP-1), IGFBP-3, and sex hormone binding-globulin (SHBG) levels; and the free androgen indexes were evaluated in 98 girls with premature pubarche [PP; prepubertal (B1; n = 32), early pubertal (B2; n = 27), midpubertal (B3; n = 23), and postmenarcheal (B5; n = 16)] and in 86 Tanner stage- and bone age-matched controls. We ascertained whether hyperinsulinemia is already present in PP girls before or during pubertal development and whether these patients show a similar pattern of growth factor secretion as normal girls. Body mass indexes did not differ significantly between patients and controls within the same pubertal stage. MSI levels showed a significant increase with pubertal onset in all subjects, as expected. Patients showed significantly higher MSI values than controls at all Tanner stages (P < 0.03, P = 0.03, P = 0.03, and P < 0.05 for B1, B2, B3, and B5, respectively); higher insulin response to glucose at B1, B2, and B3 (P < 0.03, P = 0.03, and P < 0.05, respectively); higher glucose uptake rate in peripheral tissues at B1 and B2 (P < 0.04 and P = 0.02, respectively); and a later rise in insulin sensitivity compared to controls. PP girls also showed lower IGFBP-1 levels at B1 and B5 (P < 0.01 and P = 0.02, respectively), lower SHBG concentrations at B5 (P < 0.0005), and higher free androgen indexes at B1, B3, and B5 (P < 0.01, P < 0.05, and P < 0.001, respectively) compared to controls. Among others, significant correlations between SHBG and MSI levels (r = -0.49; P < 0.0001) and between SHBG and IGFBP-1 levels (r = 0.41; P < 0.0001) were found in all subjects. Hyperinsulinemia, increased early insulin responses to glucose, increased glucose uptake rate in peripheral tissues, elevated free androgen indexes, and decreased SHBG and IGFBP-1 levels are present in most girls with PP from childhood. These findings lend strong support to the concept that PP is not a benign condition, and long term follow-up of these patients into adulthood is recommended. The possible causal role of hyperinsulinemia in adrenal and/or ovarian androgen hypersecretion remains to be established. PMID- 9215309 TI - Transcriptional regulation of prostaglandin-H synthase-2 gene in human trophoblasts. AB - Abnormal PG production by placental PG-H synthase (PGHS) is associated with preeclampsia. There are two PGHS isozymes, and their regulation in trophoblasts is presently unknown. We hypothesized that the PGHS isozymes are differentially regulated in human trophoblasts. To test this hypothesis, we transfected primary trophoblasts and JEG3 cells with promoter constructs of either PGHS-1 or PGHS-2 genes. We found that in both cell systems, the basal activity of PGHS-2 promoter was 10- to 30-fold higher than the activity of PGHS-1 promoter. In response to either 12-0-tetradecanoylphorbol-13-acetate (TPA) or 8-bromo-cAMP, we observed an increase in PGHS-2 promoter activity but no change in activity of PGHS-1 promoter. Similarly, both agents enhanced PGHS-2 expression, as well as prostaglandin E2 production. The activity of PGHS-2 promoter was potentiated by coexpression of protein kinase A and inhibited by coexpression of kinase A inhibitor. Aspirin attenuated the stimulatory effect of TPA on PGHS-2 promoter. We conclude that both PGHS-1 and PGHS-2 promoters are active in trophoblasts. The activity of PGHS-2 promoter is stimulated by either TPA or cAMP, and the stimulatory effect of TPA is attenuated by aspirin. These pathways may play a role in modulation of prostanoid synthesis by trophoblasts. PMID- 9215310 TI - Aldose reductase gene expression is increased in diabetic nephropathy. AB - Aldose reductase gene expression is increased in insulin-dependent diabetes mellitus (IDDM) with nephropathy. Epidemiology studies in patients with IDDM and noninsulin-dependent diabetes mellitus (NIDDM) are consistent with the hypothesis that a genetic factor(s) influences the risk for kidney disease of diabetes mellitus (KDDM). Aldose reductase (AR), the rate-limiting enzyme in the polyol pathway, is a potential candidate gene product. The present study explored the hypothesis that AR gene expression is increased in peripheral blood mononuclear cells obtained from patients with KDDM. We studied four groups of volunteers: group I, normal subjects; group II, IDDM without nephropathy; group III, IDDM with kidney disease; and group IV, nondiabetics with kidney disease. AR messenger ribonucleic acid was measured by a ribonuclease protection assay. The results are expressed as the mean and 95% confidence interval (CI) of the AR/beta-actin messenger ribonucleic acid molar ratios (AR/beta-actin R). Among diabetics, the AR/beta-actin R was higher in group III (0.088; CI, 0.068-0.108) than in group I (0.045; CI, 0.033-0.057; P < 0.01). There were no significant differences in age, hemoglobin A1c, or duration of diabetes between groups II and III (P = NS). The AR/beta-actin R in group III was also higher than that in group II (0.045; CI, 0.030-0.060; P < 0.01) or group IV (0.019; CI, 0.011-0.027; P < 0.001). In contrast, among nondiabetics, AR/beta-actin R values were 2-fold lower in group IV than in group I (P < 0.01). The results of this study are consistent with the hypothesis that the degree of AR gene expression modulates the risk of KDDM. PMID- 9215311 TI - Metabolism of oral glucose in pancreas transplant recipients with normal and impaired glucose tolerance. AB - To gain insight into the pathophysiology of impaired glucose tolerance in pancreas transplantation, glucose kinetics and insulin secretion were assessed after an oral glucose load in four combined pancreas-kidney recipients with impaired glucose tolerance (IPx), in five combined pancreas-kidney recipients with normal glucose tolerance, in six nondiabetic kidney transplant recipients, and in eight normal subjects employing a dual isotope technique, beta-Cell function was evaluated by calculating prehepatic insulin secretion rates, which subsequently were correlated to the ambient glucose concentrations to obtain an index of beta-cell responsiveness. Oxidative and nonoxidative glucose metabolism were assessed by indirect calorimetry. Basal insulin secretion rates, the glucose stimulated early insulin secretion rates, as well as beta-cell responsiveness were markedly reduced in IPx than in the glucose-tolerant transplant subjects. Total systemic glucose appearance was similar in the groups with apparently comparable inhibition of systemic glucose release and increase in exogenous glucose appearance. The hyperglycemic response in IPx was due to a significant reduction in the glucose disappearance rates during the first 2 h after glucose ingestion. Nonoxidative glucose metabolism increased significantly less in IPx than in glucose-tolerant groups. Glucagon secretion was less suppressed in the early part of the study in IPx, which may have contributed to the excessive hyperglycemia. In conclusion, IPx after pancreas transplantation was characterized by 1) impaired early insulin secretion, 2) reduced beta-cell responsiveness, 3) reduced glucose uptake, 4) impaired nonoxidative glucose metabolism, and 5) impaired early inhibition of glucagon secretion. PMID- 9215312 TI - Elevated serum insulin-like growth factor-binding protein 2 (IGFBP-2) and decreased IGFBP-3 in epithelial ovarian cancer: correlation with cancer antigen 125 and tumor-associated trypsin inhibitor. AB - Insulin-like growth factors (IGFs) and IGF-binding proteins (IGFBPs) recently have been shown to play a physiological role in the female genital system, including the ovarian follicular system. However, little is known about the role of the IGF system in malignant ovarian disease. To assess possible mutual correlations between alterations in circulating IGFBP profiles and tumuor markers in patients with epithelial ovarian cancer, we performed an RIA for IGFBP-2 and IGFBP-3 and a Western ligand blotting (WLB) in serum samples from 20 patients with epithelial ovarian cancer, 10 patients with benign ovarian tumors, and 8 healthy age-matched controls. The epithelial ovarian cancer group had a mean IGFBP-2 level that was 253% (RIA) and 105% (WLB) above that of controls. IGFBP-2 even correlated positively with the highly sensitive serum tumor marker, cancer antigen 125 (CA 125) (r = 0.71, P < 0.001) but not with the less sensitive tumor associated trypsin inhibitor. In contrast, serum IGFBP-3 (by RIA and WLB) was decreased in patients with ovarian cancer, and IGFBP-3 proteolytic activity was detectable in some of the patients. Neither IGFBP-3 nor IGFBP-3 proteolytic activity correlated with CA 125; but the former correlated inversely, and the latter positively, with tumor-associated trypsin inhibitor. In conclusion, IGFBP 2 levels are high in serum of epithelial ovarian cancer patients, and the increment in serum IGFBP-2 correlates positively with CA 125. Alterations in serum IGFBP-2 levels may therefore, serve as a potential additional marker for ovarian cancer. PMID- 9215313 TI - Testosterone regulates apoptosis in adult human seminiferous tubules in vitro. AB - In the present study an in vitro model was developed and characterized for evaluation of the role of apoptosis in adult human testes. The samples came from adult men undergoing orchidectomy for prostate or testicular cancer. Segments of seminiferous tubules were isolated and incubated under serum-free conditions in the absence or presence of testosterone. Apoptosis was assessed by low mol wt DNA fragmentation (185-bp multiples) by use of 3'-end-labeled DNA, in situ end labeling, and morphological detection under light and electron microscopy. During the 4-h incubation, a 15-fold increase was seen in apoptotic DNA fragmentation. The extent of low mol wt DNA showed a time-dependent increase and reached a 20 fold intensity in 24 h of incubation compared to the level at 0 h. Apoptosis was significantly suppressed by testosterone concentrations of 10(-7) and 10(-6) mol/L during the first 4 h of incubation. Apoptotic cells were identified mainly as spermatocytes and occasionally as spermatids. We conclude that apoptosis is induced in human seminiferous tubules under serum-free conditions in vitro. That this apoptosis is suppressed by testosterone indicates that testosterone in the human male is a critical germ cell survival factor. The model created in the present study provides a valuable tool for further investigation of hormonal and gene regulation of human germ cell death and survival. PMID- 9215314 TI - Negative/low expression of the Met/hepatocyte growth factor receptor identifies papillary thyroid carcinomas with high risk of distant metastases. AB - To investigate the clinical impact of Met/hepatocyte growth factor receptor (HGF R) expression in thyroid cancer we studied 163 thyroid carcinomas (129 papillary, 21 follicular, and 13 anaplastic) from patients followed-up for 25-147 months postthyroidectomy. Forty-nine thyroid adenomas were also studied. Met/HGF-R expression was evaluated by semiquantitative immunohistochemistry, measuring both the proportion (scale of 0-5) and the intensity (scale, 0-5) of stained cells and calculating a total score (scale of 0-10). Met/HGF-R was absent in the normal thyroid tissue, absent or focally expressed in follicular and anaplastic tumors, and expressed at various levels in most papillary carcinomas, including microcarcinomas. Papillary carcinomas were thus categorized as having negative/low Met/HGF-R (n = 50; total score, < or = 5) or high Met/HGF-R expression (n = 70; total score, > 5). High Met/HGF-R was inversely associated with vascular invasion (P = 0.0308), but not with other prognostic factors. Negative/low Met/HGF-R expression was the most effective predictor by multivariate Cox analysis of distant metastases (hazard ratio = 9.71; P = 0.0036), higher than extrathyroid invasion (hazard ratio = 4.25; P = 0.0181), age (< or = 45 vs. > 45 yr; hazard ratio = 3.99; P = 0.0099), and vascular invasion (hazard ratio = 3.19; P = 0.0358). These findings suggest a role for Met/HGF-R in papillary thyroid cancer and its clinical use to select patients with a high risk of distant metastases. PMID- 9215315 TI - Effect of interferon-gamma and glucose on major histocompatibility complex class I and class II expression by pancreatic beta- and non-beta-cells. AB - Surface major histocompatibility complex (MHC) class I and class II expression by pancreatic islet cells is considered a local initiator or regulator of immune processes that can lead to diabetes. Locally released cytokines, in particular interferon-gamma, are known to stimulate MHC antigen expression by islet cells. The present study quantifies MHC expression in cultured pancreatic beta- and non beta-cells from both rat and human organs. Interferon-gamma increased MHC class I expression in endocrine beta- and non-beta-cells as well as in pancreatic ductal cells. The cytokine induced a 6-fold increase in the MHC class I messenger ribonucleic acid levels in pancreatic beta-cells; this effect was 2-fold amplified in the presence of elevated glucose levels (20 mmol/L instead of 6 mmol/L). No MHC class II expression was observed in endocrine beta- or non-beta cells; human, but not rat, ductal cells exhibited MHC class II expression that increased in the presence of interferon-gamma. These data indicate that the increase in beta-cell MHC class I expression described in the pancreata of diabetic patients may result from stimulated transcription after exposure to locally released interferon-gamma and/or to a hyperglycemic state. The association of human islets with ductal cells in which MHC class II expression is stimulated by interferon-gamma makes these cells potential participants in the autoimmune process in diabetes. PMID- 9215316 TI - A novel splicing junction mutation in the gene for the steroidogenic acute regulatory protein causes congenital lipoid adrenal hyperplasia. AB - Congenital lipoid adrenal hyperplasia (lipoid CAH) is a relatively common genetic disorder of adrenal and gonadal steroidogenesis and is the most severe form of CAH. As typical affected individuals cannot produce any steroid hormones or can only produce low levels of steroid hormones in the adrenals and gonads, including glucocorticoids, mineralcorticoids, and sex steroids, a genetic defect in the cholesterol side-chain cleavage enzyme, cytochrome P450scc (CYPXIA1), has been postulated to be the cause of their insufficient production to date. Recently, Lin and co-workers proved a link between mutations of the steroidogenic acute regulatory protein (StAR) gene and the lipoid CAH phenotype. Therefore, we investigated both the cytochrome P450scc and StAR genes in a Korean family with a fairly mild form of lipoid CAH to identify the mutation(s) causing this disease. The result was that no mutations could be found in the two genes, except for a thymine (T) insertion into intron 2 of the StAR gene, 3 bp from the splice donor site of exon 2. PCR-amplified StAR genes from a normal subject and the patient were cloned into an expression vector and then introduced into COS-7 cells. Northern blot and reverse transcriptase-PCR analyses indicated that the StAR messenger ribonucleic acid derived from the vector with the normal StAR gene spliced exons 2 and 3 correctly, whereas most, but not all, StAR messenger ribonucleic acid derived from the vector with the T-inserted StAR gene could not remove intron 2. We concluded from these results that the T insertion into the StAR gene accounts for the lipoid CAH phenotype in this patient. PMID- 9215317 TI - Interleukin-6 and the interleukin-6 receptor in the human adrenal gland: expression and effects on steroidogenesis. AB - Interleukin (IL)-6 is a potent activator of the human hypothalamicpituitary adrenal axis. After chronic administration of IL-6 in humans, there is a substantial elevation of cortisol, whereas ACTH levels are blunted. Thus, we investigated whether IL-6 and/or the IL-6 receptor (IL-6R) are expressed in the human adrenal gland and whether IL-6 could cause the release of steroid hormones by a direct action on adrenal cells in primary culture. The expression of IL-6 and IL-6R was investigated with RT-PCR and immunohistochemistry, and the effects on human adrenal steroidogenesis were tested with IL-6 in vitro. To avoid effects mediated by macrophages, we depleted adrenal primary cultures from macrophages using specific mouse antihuman CD68 and sheep antimouse IgG conjugated magnetic beads. The results showed that 1): IL-6 and IL-6R are expressed in adrenal cell cultures, including all cell types and those depleted of macrophages; 2) IL-6R is mainly expressed in the zona reticularis and the inner zona fasciculata; positive signals from the zona glomerulosa and the medulla occurred in single cells; and 3) IL-6 regulates adrenal synthesis of mineralocorticoids, glucocorticoids, and androgens in vitro, dependent on time and dose, in the absence of macrophages. After 24 h, aldosterone secretion increased to 172 +/- 28% SEM, cortisol to 177 +/- 27% SEM, and dehydroepiandrosterone to 153 +/- 20% SEM of basal secretion. These findings, in combination with previous investigations, suggest that IL-6 exerts its acute action via the hypothalamus and the pituitary. In the adrenal gland, however, IL-6 seems to be a long-term regulator of stress response, integrating the responses of all cortical zones to stimuli from the immune and endocrine system. PMID- 9215318 TI - Molecular basis of nonclassical steroid 21-hydroxylase deficiency detected by neonatal mass screening in Japan. AB - Since 1989, neonatal mass screening for congenital adrenal hyperplasia (CAH) has been performed in Japan, and the frequency of the classical form of 21 hydroxylase deficiency was found to be nearly identical to that in other countries. However, it has not yet been determined whether our mass screening program can detect the nonclassical (NC) form. From 1991 to 1994, about 4,500,000 infants underwent CAH mass screening in Japan. During this period, we identified by screening 2 siblings and 2 unrelated patients who had mild elevation of serum 17-hydroxyprogesterone levels at 5 days of age, but who revealed no symptoms of CAH. They were diagnosed as having probable NC steroid 21-hydroxylase deficiency. To clarify the molecular basis of NC CAH detectable by neonatal screening in Japan, the steroid 21-hydroxylase (CYP21) genes from these cases were analyzed. The 2 siblings (patients 1 and 2) had I172N and R356W mutations in 1 allele and in the other allele had local gene conversion, including the P30L mutation in exon 1. Patient 3, who was unrelated, had gene conversion encoding the same P30L mutation in 1 allele and in the other allele had an intron 2 mutation (668-12 A- >G), causing aberrant ribonucleic acid splicing, and the R356W mutation. Patient 4, also a compound heterozygote, had the R356W and 707del8 mutations. The estimated rate of detection of the NC form by mass screening (1:1,100,000) seemed low compare to the established detection rate for the classical form (1:18,000). As all of our 4 patients were compound heterozygotes with at least 1 allele bearing 1 or more mutations associated with classic CAH, it may be difficult to detect NC cases carrying only NC-associated alleles using our current neonatal mass screening methods. PMID- 9215319 TI - Clinical, radiological and pathological features of patients with Rathke's cleft cysts: tumors that may recur. AB - Rathke's cleft cysts are cystic sellar and suprasellar lesions, characteristically lined by a single layer of ciliated cuboidal or columnar epithelium. In contrast, craniopharyngiomass, which are also cystic sellar and suprasellar lesions, are characteristically lined by stratified squamous epithelium with keratinization on a layer of connective tissue. The usual management recommended for Rathke's cleft cysts is simple surgical drainage with partial excision of the cyst wall. Recurrences of these cysts reportedly have been very rare. This retrospective study presents the details of 12 patients (6 females; median age 30 yr, range 21-58 yr) with Rathke's cleft cyst, referred to our department over a 15-yr period (1981-1996), an unusual feature being the recurrence of 4 (33%) of these lesions. Clinical, endocrine, radiological, surgical (10 transsphenoidal; 2 transcranial), and pathological details were recorded. Nine out of 12 patients (75%) were symptomatic; visual symptoms were the commonest, and 8 had visual field defects. The median duration of symptoms was 12 months (range 3-24 months). Three patients (25%) had panhypopituitarism, 2 of whom also had diabetes insipidus (17%). The cysts varied in size from 6 mm to 50 mm, 1 being entirely suprasellar. There were no pathognomonic clinical or radiological features to differentiate them from other pituitary lesions, although the presence of diabetes insipidus in 2 patients suggested that the lesion was not a pituitary adenoma. A definite histological diagnosis was possible in 8 patients; in 4, the diagnosis was presumptive. The median duration of follow-up was 30 months (1-168 months). Four patients (33%) showed reexpansion at 3, 6, 48, and 48 months after initial surgery, 3 of whom were symptomatic and required repeat surgery. Two of these patients were given postoperative external beam pituitary radiotherapy. Apparent recurrence of Rathke's cleft cysts after initially successful surgery in our series was higher than suggested by previous reports, and thus, long-term follow-up with pituitary imaging and neuroophthalmological assessment is essential. There are no specific characteristics of the cyst that predict recurrence. Ideal management of these cysts is unclear, but aspiration, followed by extensive excision of the cyst wall when possible, seems to be the best initial option. For recurrent symptomatic tumors, surgical resection is the treatment of choice. Considering the high recurrence rate with residual structural and functional dysfunction, the role of radiotherapy in preventing recurrence of these cysts needs careful evaluation with a larger study with a longer follow-up period. PMID- 9215320 TI - Effects of a two-week physiological dehydroepiandrosterone substitution on cognitive performance and well-being in healthy elderly women and men. AB - The levels of dehydroepiandrosterone (DHEA) and its sulfate ester DHEAS decrease with age after a peak around 25 yr. Animal studies as well as the first studies in humans have generated the idea that DHEA replacement in elderly subjects may have beneficial effects on well-being and cognitive functions. In the present experiment 40 healthy elderly men and women (mean age, 69 yr) participated in a double blind, placebo-controlled DHEA substitution study. For 2 weeks subjects took 50 mg DHEA daily, followed by a 2-week wash-out period and a 2-week placebo period. The treatment sequence was randomized in a cross-over design. After 2 weeks of DHEA or placebo, psychological and physical well-being as well as cognitive performance were assessed using several questionnaires and neuropsychological tests. All subjects had low DHEAS baseline levels. DHEA substitution lead to a 5-fold increase in DHEAS levels in women (from 0.67 +/- 0.1 to 4.1 +/- 0.4 micrograms/mL; P < 0.001) and men (from 0.85 +/- 0.1 to 4.5 +/ 0.4 micrograms/mL; P < 0.001). DHEA, androstenedione, and testosterone levels also increased significantly in both sexes (all P < 0.001). No significant changes were observed in insulin-like growth factor I or insulin-like growth factor-binding protein-3 levels. DHEA replacement had no strong beneficial effect on any of the measured psychological or cognitive parameters. Only women tended to report an increase in well-being (P = 0.11) and mood (P = 0.10), as assessed with questionnaires. They also showed better performance in one of six cognitive tests (picture memory) after DHEA. However, after Bonferroni alpha adjustment, this difference was no longer significant. No such trend was observed in men (P > 0.20). Likewise, no beneficial effects of DHEA substitution could be observed in any of the other tests of the neuropsychological test battery in either sex (all P > 0.20). In conclusion, the present data do not support the idea of strong beneficial effects of a physiological DHEA substitution on well-being or cognitive performance in healthy elderly individuals. PMID- 9215321 TI - Synthesis of IGFBP-3 fragments in a baculovirus system and characterization of monoclonal anti-IGFBP-3 antibodies. AB - IGFBPs play an important role in IGF biological actions by modulating IGF binding to its receptors. The major IGFBP in serum is IGFBP-3, which transports 70-90% of the circulating IGFs. In target cell systems, it sequesters IGFs and inhibits their hormonal actions, but may potentiate IGF activity or exert IGF-independent effects under specific conditions. IGFBP-3 can be modified by IGFBP-3 proteases, which degrade it into smaller fragments. IGFBP-3 fragments generated by proteolysis have reduced affinity for IGFs, thereby modifying IGF action. To study IGFBP-3 fragments in vivo and in vitro, we constructed six different IGFBP 3 fragments by use of a baculovirus expression system and generated 8 different monoclonal IGFBP-3 antibodies. Based on the known cleavage sites of IGFBP-3 for PSA, MMPs, and the predicted plasmin cleavage sites, we expressed a N-terminal IGFBP-3(1-97) fragment and a C-terminal IGFBP-3(98-264) fragment. By stepwise truncation from the C-terminal end, we created IGFBP-3(98-232), IGFBP-3(98-206), IGFBP-3(98-179), and IGFBP-3(98-159). A strong recognition of the C-terminus and the intermediate parts of IGFBP-3 by six antibodies was found. Four of these mAbs were able to recognize the intermediate fragment alone. Two mAbs were found to immunoreact only with the N-terminal IGFBP-3 fragment and two additional mAbs recognized the N- as well as the C-terminal parts and lacked immunoreactivity for the intermediate part of IGFBP-3. The 15 kDa IGFBP-3 fragment resulting from plasmin digestion was found to only react with N-terminal antibodies, while the 29 kDa fragment in pregnancy serum reacted with both N- and C-terminal antibodies. Thus, these mAbs will be useful tools to determine whether IGFBP-3 fragments found in vivo derive from either the N- or C-terminal domains of IGFBP 3. PMID- 9215322 TI - Expression of estrogen receptor-beta in human breast tumors. AB - The expression of a recently described novel estrogen receptor, ER-beta, was detected in several human breast tumor biopsy samples and several human breast epithelial cell lines using reverse transcription and polymerase chain reaction (RT-PCR) analysis. Cloning and sequencing of the PCR product from a breast tumor confirmed the identity of the sequence with that of the ER-beta mRNA previously reported in human testis. The expression of ER-beta was not correlated with that of ER-alpha, and both ER-alpha positive and ER-alpha negative cell lines expressed ER-beta mRNA. However, some breast tumors and some cell lines coexpress ER-beta and ER-alpha mRNA. Our data support a possible role for ER-beta in human breast cancer. PMID- 9215323 TI - Atrial natriuretic factor and digoxin-like immunoreactive factor in diabetic patients: their interrelation and the influence of the autonomic nervous system. PMID- 9215324 TI - Octreotide and lanreotide treatment in active acromegaly. PMID- 9215325 TI - Paralysis: the leading presentation for primary aldosteronism in Taiwan. PMID- 9215326 TI - Alcohol, estrogens, and breast cancer. PMID- 9215327 TI - Effects of a restraint reduction intervention and OBRA '87 regulations on psychoactive drug use in nursing homes. AB - OBJECTIVES: To describe the changes in psychoactive drug use in nursing homes after implementation of physical restraint reduction interventions and mandates of the Omnibus Budget Reconciliation Act of 1987 (OBRA '87). METHODS: A secondary analysis was conducted using data from a controlled clinical trial that took place in three nursing homes: a control home, one that received an educational intervention, and one that received an educational/consultation intervention. All three homes were influenced by the OBRA mandates. Complete pre- and 6 months' post-intervention data on use of psychoactive drugs and physical restraints were available for 446 resident subjects. Changes were first analyzed with the resident subjects as the unit of analysis and then using the nursing home ward (n = 16) as the unit of analysis. RESULTS: While physical restraint use declined in the home that received the educational/consultation intervention, neither neuroleptic nor benzodiazepine use increased in any of the homes after the interventions. The percentage of residents taking neuroleptics declined in the control home (18.6% to 11.3%, P = .014). Benzodiazepine use, which was more prevalent than described previously in the literature, declined in all three homes (P < .001). Of those residents whose physical restraints were discontinued, only 2% were started on neuroleptics. When the effect of OBRA mandates on appropriateness of neuroleptic use was examined, the percentage of residents on neuroleptics who lacked an OBRA-approved indication declined from 21.3% to 14.6% in the total sample, and from 39.9% to 8% in the control home. CONCLUSIONS: Interventions to reduce physical restraint did not lead to an increase in psychoactive drug use; further, reduction in both can occur simultaneously. OBRA mandates regarding psychoactive drug use were not uniformly effective, but appear, at minimum, to have increased awareness of the indications for neuroleptics. PMID- 9215328 TI - Deaths caused by bedrails. AB - OBJECTIVES: To determine how bedrails cause death in order to suggest clinical and ergonomic changes to prevent such deaths and to promote research to improve the use and design of bed systems. DESIGN: A review of reports of adult deaths and injuries from bedrails contained in the United States Consumer Product Safety Commission Death Certificate File and its Reported Incidents File and its National Injury Information Clearinghouse Accident Investigations from 1993 to 1996. Deaths involving the use of vest restraints were excluded. We reconstructed, reenacted, and have graphically depicted major patterns of deaths. A review of the literature to 1966 was also done. RESULTS: The 74 deaths described are categorized into three types: (1) 70% were entrapments between the mattress and a rail so that the face was pressed against the mattress, (2) 18% were entrapment and compression of the neck within the rails, and (3) 12% were deaths caused by being trapped by the rails after sliding partially off the bed and having the neck flexed or the chest compressed. CONCLUSIONS: Deaths from bedrails are underrecognized and preventable clinical events that can occur in any medical setting. Preventing these events will require a unified redesign of the relationships between rails, mattresses, and beds, which are now often assembled and used as separate products. Clinicians can prevent many of these deaths by using bedrails much more judiciously, confirming the proper relationships between beds, rails and mattresses, and using alarms. PMID- 9215329 TI - Reduced systemic arterial compliance is associated with left ventricular hypertrophy and diastolic dysfunction in older people. AB - OBJECTIVES: To study the relationship between left ventricular diastolic function and systemic arterial compliance in the older population. DESIGN: Cross-sectional survey. PARTICIPANTS: A total of 67 older volunteer participants (aged 67 +/- 5.4 years). MEASUREMENTS: Systemic arterial compliance (SAC) was measured using applanation tonometry and aortic velocimetry, and diastolic function was assessed using Doppler filling. Left ventricular mass was determined echocardiographically. RESULTS: There were significant univariate correlations between diastolic filling, as measured by E/A ratio, systemic arterial compliance (0.34, P < .01), and left ventricular mass (-0.41, P < .001). In multiple regression analysis, using diastolic filling as the dependent variable and heart rate, age, left ventricular mass corrected for body surface area, systolic and diastolic blood pressures, and arterial compliance as independent variables, the major determinants of diastolic filling were heart rate, left ventricular mass, and diastolic blood pressure. Arterial compliance did not make a significant independent contribution. CONCLUSION: This study demonstrates a positive relationship between diastolic filling and arterial compliance in the older population. However, in multiple regression analysis, heart rate, diastolic blood pressure, and left ventricular mass were the independent predictors of diastolic filling (E/A), whereas arterial compliance was not. These findings imply that therapeutic modulation of aortic stiffness would not, of itself, contribute to improvement in diastolic function. PMID- 9215330 TI - Use of digoxin, diuretics, beta blockers, angiotensin-converting enzyme inhibitors, and calcium channel blockers in older patients in an academic hospital-based geriatrics practice. AB - OBJECTIVE: To investigate the prevalence of and indications for digoxin use and the prevalence of beta blocker and calcium channel blocker use in older patients with previous myocardial infarction or coronary artery disease (CAD), and the prevalence of use of diuretics, beta blockers, angiotensin-converting enzyme (ACE) inhibitors, and calcium channel blockers in older patients with hypertension in an academic hospital-based geriatrics practice. DESIGN: A retrospective analysis of charts from 528 unselected older patients, seen from June 1995 through July 1996 at an academic hospital-based geriatrics practice, was performed to investigate the prevalence of digoxin use and indications for digoxin use, the prevalence of beta blocker and calcium channel blocker use in older patients with previous myocardial infarction or coronary artery disease (CAD), and the prevalence of use of diuretics, beta blockers, angiotensin converting enzyme (ACE) inhibitors, and calcium channel blockers in older patients with hypertension. SETTING: An academic hospital-based, primary care geriatrics practice staffed by fellows in a geriatrics training program and full time faculty geriatricians. PATIENTS: A total of 416 women and 112 men, mean age 81 +/- 8 years (range 58 to 101), were included in the study. MEASUREMENTS AND MAIN RESULTS: Ninety-two of the 528 patients (17%) were taking digoxin. Recorded indications for digoxin were atrial fibrillation with or without congestive heart failure (CHF) in 39% of patients, CHF with sinus rhythm and abnormal left ventricular ejection fraction (LVEF) in 18% of patients, a clinical assessment of CHF with sinus rhythm and no recorded measurement of LVEF in 20% of patients, paroxysmal atrial fibrillation in 14% of patients, and coronary artery disease (CAD) in 9% of patients. Of 121 patients with previous myocardial infarction, 23 (19%) were prescribed beta blockers, and 54 (45%) were taking calcium channel blockers. Of 173 patients with CAD, 41 (24%) were treated with beta blockers, and 79 (46%) were taking calcium channel blockers. LVEF was not recorded in the charts of 90 of 121 patients (74%) with prior myocardial infarction and of 125 of 173 patients (72%) with CAD. Of 480 older patients with hypertension, 154 (37%) were treated with diuretics, 55 (13%) were treated with beta blockers, 160 (38%) were treated with ACE inhibitors, and 197 (47%) were treated with calcium channel blockers. CONCLUSIONS: In 528 older patients seen in an academic hospital-based geriatrics practice, the prevalence of digoxin use was 19%. Appropriate indications for digoxin were documented clearly in the charts of 53 of 92 patients (57%). Calcium channel blockers were used more often than beta blockers in patients with previous myocardial infarction or CAD. Calcium channel blockers were the most frequently used antihypertensive drugs. PMID- 9215331 TI - Glucose metabolism in older adults: a study including subjects more than 80 years of age. AB - OBJECTIVE: This study was undertaken to understand the dynamics of glucose metabolism in healthy non-diabetic subjects older than age 80 (old-old) compared with subjects aged 61 to 79 (young-old), as well as to compare healthy older subjects with impaired glucose tolerance (IGT) with older subjects with normal glucose tolerance (NGT). DESIGN: A cross sectional, observational study. SETTING: A university hospital clinical research center. PARTICIPANTS: There were 28 community-dwelling adults, 10 older than age 80 and 18 aged 61 to 79. Thirteen of these people had NGT and 15 had IGT. Subjects were not taking any medication that interfered with glucose tolerance. MEASUREMENTS: Status of glucose tolerance was determined by an oral glucose tolerance test categorized as NGT or IGT according to WHO criteria. Insulin sensitivity (SI) and glucose effectiveness (SG) were assessed using a tolbutamide-assisted intravenous glucose tolerance test (IVGTT). The data were analyzed using the Minmod modeling program. Glucose tolerance (K(g)) and the acute insulin response to glucose (AIRg) were calculated from the IVGTT. RESULTS: There were no significant differences between the young-old and old-old in body mass index or in plasma glucose, insulin, or C-peptide levels in the fasting state or during the OGTT. Values for K(g), SI, SG, and AIRg from the IVGTT were similar in the two age groups. When the subjects were classified by glucose tolerance status, the subjects with NGT had age, gender, and body mass index similar to the subjects with IGT. Older people with IGT had a lower K(g) and tended to have higher fasting glucose and similar fasting insulin compared with people with NGT. IGT subjects had lower SI and tended to have lower SG. The AIRg in IGT subjects tended to be low rather than high when compared with older people with NGT. CONCLUSION: Otherwise healthy adults more than 80 years of age have measures of glucose metabolism similar to people aged 61 to 79. The presence of IGT in older adults is associated with insulin resistance, regardless of patient age. We hypothesize that the lack of pancreatic islet compensation for insulin resistance may contribute to impaired glucose tolerance in older adults. PMID- 9215332 TI - Surrogate and physician understanding of patients' preferences for living permanently in a nursing home. AB - OBJECTIVE: To evaluate patients' willingness to live permanently in a nursing home and surrogate and physician understanding of that preference. DESIGN: Evaluation of cross-sectional interview data from a cohort study. SETTING: Five academic medical centers. PARTICIPANTS: Seriously ill hospitalized adults enrolled in the Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments (SUPPORT). MEASUREMENTS: Patients' willingness to live permanently in a nursing home was measured on a 5-point scale ranging from "very willing" to "rather die." Ordinal logistic regression was used to identify patient demographic and clinical characteristics associated with this preference. Surrogate and physician perceptions of patient preferences were compared with patients' responses, and factors associated independently with surrogate and physician understanding of patient preference were identified. RESULTS: Of 9105 patients, 3262 (36%) provided responses to the study question: 7% were "very willing" to live permanently in a nursing home, 19% "somewhat willing," 11% "somewhat unwilling," 26% "very unwilling," and 30% would "rather die." Older age was associated independently with less willingness to live permanently in a nursing home (odds ratio [OR] = .90 per decade; 95% confidence interval [CI]: 0.85, 0.96). Patients with more education (OR = 1.03 per year; 95% CI: 1.00, 1.05) and more disabilities (OR = 1.05 per disability; 95% CI: 1.01, 1.09), and black patients (OR = 1.46 compared with white patients; 95% CI: 1.20, 1.76) were more willing to live in a nursing home. Surrogates understood 61% of patients' nursing home preferences but identified only 35% of patients who were willing to live permanently in a nursing home. Physicians identified 18% of patients willing to live permanently in a nursing home. CONCLUSION: Patient attitudes about living permanently in a nursing home can be elicited, cannot be reliably predicted from demographic and clinical variables, and are frequently misunderstood by surrogates and physicians. Elicitation of patient preferences regarding permanent nursing home placement should be explored before patients become unable to participate in decision making in order to enhance the concordance of patient preference with the way they spend the end of their lives. PMID- 9215333 TI - Incidence of anemia in older people: an epidemiologic study in a well defined population. AB - OBJECTIVE: To assess the incidence and clinical spectrum of anemia among older people. DESIGN: Inception cohort assembled and followed by medical records linkage until death or last clinical contact through January 1994. SETTING: Population-based study in Olmsted County, Minnesota. PARTICIPANTS: All 618 Olmsted County men and women aged 65 years or more with anemia by World Health Organization criteria that was first recognized in 1986. MEASUREMENTS: Age- and sex-adjusted incidence rates, corrected for prevalent anemia, and survival estimates using the Kaplan-Meier method, with calculation of standardized mortality ratios for specific causes of death. RESULTS: The corrected annual incidence of anemia rose with age, and rates were higher in men (90.3 per 1000; 95% CI, 79.2-101.4) than women (69.1 per 1000; 95% CI, 62.3-75.8). In 465 cases (75%), anemia was detected in conjunction with a hospitalization, but admission was due to anemia in only 57 instances. Half of the cases were caused by blood loss, two-thirds of these as a result of surgery. The cause of anemia was uncertain in 102 cases (16%). One-third of the patients were transfused with a median of 3 units each. Overall survival was worse than expected but was better among those with anemia caused by blood loss. Mortality attributable to malignancy, mental disorders, circulatory and respiratory diseases, ill-defined conditions, and injuries was significantly increased among these older patients with anemia. CONCLUSIONS: The incidence of anemia among older people is 4 to 6 times greater than that suspected clinically, rises with age, and is higher in men than in women. The apparent cause in half the cases is blood loss. Even mild anemia is associated with reduced survival, especially during the first year, but this could relate to underlying comorbid conditions. PMID- 9215334 TI - The Barthel activities of daily living index: self-reporting versus actual performance in the old-old (> or = 75 years). AB - BACKGROUND AND PURPOSE: The Barthel Index for assessing activities of daily living (ADL) was developed particularly for young stroke patients, but it now has a wider application in the geriatric assessment profile. This study tests the validity of the Barthel Index by self-report in the old-old (> or = 75 years). If more than 10% of the studied population assessed themselves incorrectly (> or = 15-point discrepancy), the test may have limitations. We set out to try to quantify and explain this discrepancy. METHODS: During a 3-month period, we tested 126 old-old patients, both geriatric medical inpatients and subjects from the community, in a cross-sectional study. Using the Barthel Index, their functional status was assessed by self-report and by observation of performance. A measure of the magnitude of discrepancy between the two methods (discrepancy score) was calculated as the difference between the self-report and performance total scores. RESULTS: Comparing the self-report with actual ADL performance scores, the mean score for self-report was higher (90 vs 88). There was a low Kappa score in all areas of the scale (range 0.103-0.398). Twenty of the 126 patients (15.9%) scored 15 or more points in the discrepancy score. By running a multiple linear regression, we were able to explain only 21% of the variance in the discrepancy score (R2 = .21). Significant explanatory variables were the presence of cognitive impairment, source of patients from acute geriatric ward, and age (very old > or = 85 years). CONCLUSION: For the purpose of this study, use of the Barthel Index by self-reporting was found to have its limitations in the old-old (> or = 75 years), particularly with regard to the very old (> or = 85 years) medical geriatric inpatients. Therefore, we suggest that the older people may have to be assessed by the rehabilitation services using a performance based measure or a different self-report test for documenting their activities of daily living, bearing in mind that self-reported and performance-based measures capture physical abilities differently. PMID- 9215335 TI - Cross-validation of anthropometric and bioelectrical resistance prediction equations for body composition in older people using the 4-compartment model as a criterion method. AB - OBJECTIVE: To cross-validate existing equations in the literature for their accuracy and precision for estimating body fat in older people from anthropometric measures and height2/resistance, using the 4-compartment model as a criteria method, and to propose new practical equations with improved accuracy and precision. DESIGN: Measurement of body composition in a cross-sectional cohort of healthy men and women and comparison by cross-validation techniques against existing prediction equations. SETTING: The study was performed on subjects admitted to a General Clinical Research Center. PARTICIPANTS: The subjects were 41 healthy women (68.2 +/- 6.6 years; 64.1 +/- 10.2 kg) and 41 healthy men (70.2 +/- 7.0 years; 74.9 +/- 11.0 kg). MEASUREMENTS: The criteria method for total body composition was the 4-compartment model based on measurement of total body density by underwater weight, total body water by isotope dilution, and total bone mineral from dual energy X-ray absorptiometry. The other techniques examined for accuracy and validity were body fat estimates derived by skinfolds using the Durnin and Womersley Equations; waist circumference and age using the Lean Equations; and bio-electrical resistance using five published equations, including two derived in the older population. RESULTS: When compared with data derived from the 4-compartment model, the skinfold equation of Durnin and Womersley was cross-validated successfully in women but not in men. The Baumgartner equation was the only bioelectrical resistance equation that met the criteria for successful cross-validation in men and women, although in women the intercept (4.0 +/- 2.1 kg) was close to significantly different from zero (P = .06). Error in the estimates of body fat using the Durnin and Womersley and the Baumgartner equations was significantly and inversely related to fat mass (r = -.39 to r = -.56). In our data, the significant predictors of fat mass were hip circumference, calf skinfold, gender, body weight, height2/resistance, and biceps skinfold, explaining 84% of the variance. CONCLUSIONS: The Durnin and Womersley equation is accurate for women and the Baumgartner equation is accurate for both men and women if a correction of +4 kg is made in women; however, for both equations the error in the estimate is inversely related to fat mass. We suggest new anthropometric equations for estimating body fat in older people, which may improve accuracy and precision. The new equations will need to be tested in independent cross-validation studies. PMID- 9215337 TI - Osteoarthritis of the knee: report of a Task Force of the International League of Associations for Rheumatology and the Osteoarthritis Research Society. PMID- 9215336 TI - The burden of Parkinson's disease on society, family, and the individual. AB - OBJECTIVE: To examine the burden of Parkinson's Disease (PD) on society, family, and the individual. SETTING: In-home interviews in Central North Carolina. DESIGN: A cross-sectional, descriptive study. PARTICIPANTS: A total of 109 people with PD. MEASURES: Standard instruments used to assess income, health status, health-related costs, and household activities. SAMPLE: The sample was weighted toward individuals who were within the first 5 years of post-PD diagnosis. RESULTS: The total per capita societal burden was approximately $6000 per year, the greatest single element of which was compensation for earnings loss for those less than age 65. Government insurance covered 85% of our sample. The largest components of family burden were the burden of providing informal caregiving and that of earnings loss. Spouses providing informal care did so a mean of 22 hours per week. Compared with a random sample of older people, our respondents spent much less time on house and yard work. CONCLUSION: The direct costs of the disease reflect a small portion of the burden. The hidden costs, in the form of lost wages, informal care, and changing roles are substantial. Their magnitude is even more important when we consider that the family generally lives on a fixed income, and the caregiver is an older aged spouse. Medical practitioners will best be able to intervene if they look holistically at the patient with this disease. When treating symptoms themselves, practitioners need to be aware of the high level of pain, fatigue, and depression associated with PD, even in the early stages. The results demonstrate clearly that family relationships are affected early, indicating the importance of providing early referrals to services such as home health, social workers/counseling, and well as PD support groups. PMID- 9215338 TI - Determinants of initiation and suboptimal use of anti-ulcer medication: a study of the Quebec older population. AB - OBJECTIVES: To describe the use of anti-ulcer medication in the Quebec older population; to examine determinants of initiation, suboptimal use, and switches between products. DESIGN: Population-based retrospective cohort study. SETTING: Universal health program for older adults in Quebec. PARTICIPANTS: 5000 users and 5000 non-users of anti-ulcer medications were selected randomly. Use was defined as the presence in the 1991 prescription database of an anti-ulcer prescription. Among users, 1697 (34%) were new users and were considered as the exposure group. Subjects were followed for 365 days after inclusion. MEASUREMENTS: Measured were patient's age, gender, prescribed duration of anti-ulcer medication, concomitant medications, and gastrointestinal diagnostic procedures. RESULTS: A total of 17% of new users had unusually short courses; 18% were long-term users. There was no difference in duration for omeprazole compared with other anti-ulcer medications. First-time use of NSAIDs was the strongest predictor of initiation of anti-ulcer medication (odds ratio = 3.21; 95% CI, 2.66-3.88). Twenty-six percent of users switched brands. Only 9.5% of new users underwent a diagnostic procedure before initiation of therapy, and 49% of long-term users ever underwent such procedure. CONCLUSION: Despite a relatively homogeneous recommended duration of therapy, patterns of use of anti-ulcer medication among older people are highly variable, and treatment is often not accompanied by a diagnostic procedure. PMID- 9215339 TI - Comparison of subcutaneous and intravenous administration of a solution of amino acids in older patients. AB - OBJECTIVE: To compare the plasma amino acid (AA) concentrations obtained by the infusion of an AA solution (660 mOsm/L, pH 7) using the subcutaneous (SC) with that using the intravenous (i.v.) route in older patients. DESIGN: A prospective, randomized, cross-over study. SETTING: A hospital geriatric ward. PARTICIPANTS: Six patients with a mean age of 84 years. MEASUREMENTS: The infusion of the AA solution (IV or SC) lasted 6 hours. Blood was sampled at 0, 2, 4, 6, 8, 10, 14, 18, and 24 hours from the start of the infusion to determine plasma AA level by the phenyl-isothiocyanate method. RESULTS: Compared with baseline values, plasma AA concentrations increased to a significantly higher level from the second to the tenth hour and from the second to the fourteenth hour during i.v. and SC infusions, respectively. Plasma AA levels did not differ between the two routes. SC infusion was well tolerated. CONCLUSION: Assuming that nutritional sufficiency is the desired result of plasma AA infusion, we conclude the SC route is well tolerated and offers the possibility of nitrogen supplementation for older patients over short periods of time, when oral protein intake is transiently insufficient or impossible, as a way to limit, but not to treat, protein-energy malnutrition. PMID- 9215341 TI - Oropharyngeal candidosis in the older patient. AB - Colonization of the oral and pharyngeal regions by Candida spp., particularly C. albicans, is extremely common in humans, particularly in early and late life. A variety of local and systemic conditions predispose the transformation of the benign colonization to a pathological state, which may have severe local or serious systemic consequences. The finding of oropharyngeal candidosis in an older patient, therefore, merits investigation of the likely host factors responsible for the organism adopting its pathogenic behavior. This paper provides non-dental clinicians managing older patients a review of the clinical characteristics, risk factors, diagnosis, and management of oropharyngeal candidosis in older adults. PMID- 9215342 TI - Structural modifications of proteins during aging. PMID- 9215343 TI - Post-stroke rehabilitation guidelines. The Clinical Practice Committee of the American Geriatrics Society. PMID- 9215344 TI - Rational allocation of medical care: a position statement from the American Geriatrics Society. AGS Ethics Committee. PMID- 9215345 TI - Allocating medical resources: recommendations for a professional response. PMID- 9215346 TI - Arterial compliance, systemic blood pressure, and diastolic dysfunction in older adults. PMID- 9215347 TI - Some "depressive" symptoms may not imply depression. PMID- 9215348 TI - Charles Bonnet syndrome, insight and cognitive impairment. PMID- 9215349 TI - Charles Bonnet syndrome and early dementia. PMID- 9215350 TI - Charles Bonnet syndrome. PMID- 9215351 TI - Treatment of pressure ulcers with nitropaste. PMID- 9215352 TI - Are congenital malformations in older people underdiagnosed? PMID- 9215353 TI - Asset and Health Dynamics Among the Oldest Old (AHEAD): initial results from the longitudinal study. Introduction. PMID- 9215354 TI - Asset and Health Dynamics Among the Oldest Old: an overview of the AHEAD Study. AB - This chapter provides background information for the study of Asset and Health Dynamics Among the Oldest Old (AHEAD), a prospective panel survey of persons born in 1923 or earlier who were residing in the community at the time of the 1993 baseline. Interviews were sought with both spouses in married households, and an overall total of 8,222 were completed. We review the interdisciplinary scientific issues that motivated the study, describe the fundamental design decisions that structured AHEAD, and summarize the content in the core and experimental modules. The study provides unusually detailed data on cognition, family structure and transfers, and assets. Data are presented on sample selections, response rates, and oversamples of minority groups. Basic descriptive data on the demographic, health, and socioeconomic attributes of respondents also are presented. Plans for future waves of AHEAD are described, including a next-of-kin interview for decreased respondents. PMID- 9215355 TI - A comparative analysis of ADL questions in surveys of older people. AB - This article describes questions designed to assess limitations with respect to activities of daily living (ADLs) that were asked on the first wave of the AHEAD study, and it assesses their cross-sectional measurement properties. It also provides comparisons between those questions and parallel questions that have been asked on two other surveys of the elderly population in the United States: the 1984 Supplement on Aging (SOA) to the National Health Interview Survey and the screener for the 1982 National Long Term Care Survey (NLTCS). It also compares a single item from the 1990 Census. It then compares the ways in which the same individuals answer these different versions of ADL questions, using data from subsamples of the AHEAD respondents who were also asked the SOA, NLTCS, or Census questions. The analysis shows that there is a substantial amount of measurement error in the answers to ADL questions, and it suggests that this is a major contributor to apparent improvements and declines in functional health observed in longitudinal data. PMID- 9215356 TI - Measures of cognitive functioning in the AHEAD Study. AB - Decline in cognitive functioning and onset of cognitive impairment are potentially important predictors of elderly persons needing informal assistance and formal health care. This article describes the measures of cognitive functioning that were developed for the Asset and Health Dynamics Among the Oldest Old (AHEAD) study of some 6,500 Americans aged 70 years and older. The study was designed to investigate the impact of health on disbursement of family and economic resources. Evaluation of the cognitive measures in terms of psychometric properties and missing data, telephone administration, and formation of an aggregate index is encouraging. Their construct validity is evidenced by their correlations with sociodemographic characteristics and health indicators that replicate existing findings as well as by their prediction of IADL and ADL functioning that are consistent with theory. PMID- 9215357 TI - The structure of health status among Hispanic, African American, and white older adults. AB - Activities of daily living (ADLs), instrumental ADLs, and disability markers have traditionally been the most common indicators of functional status. The study on Asset and Health Dynamics Among the Oldest Old (AHEAD) is used to replicate a five-dimensional measurement model composed of these observable indicators among the older adult self-respondents. The items available to measure upper body disability were found wanting, but the lower body disability, and the basic, household, and advanced ADL constructs were confirmed. Analyses of the measurement model separately among subgroups of women, men, Hispanics, Mexican Americans, African Americans, and Whites found no meaningful differences. Two structural models linking the lower body disability, and the basic, household, and advanced ADL constructs to perceived health and depression were also replicated among the older adult self-respondents, as well as separately among African Americans and among Whites. These models reaffirmed the dominant role of lower body disability on the everyday activities of older adults, and on their perceived health and depression. PMID- 9215358 TI - Race, socioeconomic status, and health: accounting for race differences in health. AB - This article uses the Asset and Health Dynamics Among the Oldest Old (AHEAD) study to examine the extent to which observed differences in the prevalence of chronic conditions and functional limitations between Black and White adults (aged 70+) in the United States can be attributed to differences in various aspects of socioeconomic status (SES) between these groups. We use linear and logistic regression techniques to model the relationships between health outcomes and SES. Our findings indicate that race differences in measurable socioeconomic characteristics indeed explain a substantial fraction, but in general not all, of Black/White differences in health status. While our findings do not suggest that low SES directly "causes" poor health, any more than being Black does so, they do suggest that research and policy intended to address the deficit in health status among Blacks (when compared to Whites) in the U.S. would be well-served to begin with the deficit in wealth, education, and other SES measures. PMID- 9215359 TI - Wealth inequality among older Americans. AB - Using the AHEAD study, this article examines the wealth distribution among American households with a member at least 70 years old. Household wealth is quite unevenly distributed among older American households. Those households in the top 10th percentile of the wealth distribution have 2,500 times as much wealth as those at the lowest 10th percent. This sharp wealth disparity relative to income dispersion is the dominant reason why older minority households have accumulated so little wealth compared to White households. Wealth varies by a factor of seven to one when both spouses are in poor health compared to when they say that they are in excellent health. Finally, AHEAD data on bequest intentions suggest a bifurcated bequest motive. Most older households plan to bequeath a modest financial inheritance, but about one-quarter expect to leave inheritances worth $100,000 or more. PMID- 9215360 TI - Transfer behavior within the family: results from the Asset and Health Dynamics Study. AB - When individuals fall on hard times, can they rely on their family for financial support? In view of proposed reductions in public assistance programs, it is important to understand the mechanisms through which families provide support for their members. In this article we provide evidence that intrafamily transfers are compensatory, directed disproportionately to less well-off members. In a given year, adult children in the lowest income category are 50 percent more likely to receive a financial transfer from their parents, and on average they receive over $300 more than their siblings who are in the highest income category. The dataset used in the new Asset and Health Dynamics (AHEAD) study contains information on all children in the family; therefore, we are able to estimate models that control for unobserved differences across families. Our results are robust to these specifications. In addition, we do not find evidence that parents provide financial assistance to their children in exchange for caregiving. PMID- 9215361 TI - Patterns of in-home care among elderly black and white Americans. AB - This study examines the use of informal and formal sources of care by elderly Black and White Americans (n = 2,847) who are functionally impaired and noninstitutionalized. The data are from the Asset and Health Dynamics Among the Oldest Old (AHEAD) study. Detailed baseline characteristics are provided and logistic regressions are used to assess the likelihood of (a) receiving in-home assistance from any source, (b) using any informal sources of in-home care, (c) using any formal sources, and (d) using formal sources of in-home care with informal sources of home care. Results of the logistic regressions indicate that, compared to Whites, Black elders were less likely to receive assistance and to use informal sources of home care. PMID- 9215362 TI - The division of family labor: care for elderly parents. AB - We consider the division of caregiving efforts among the children of older, functionally limited parents. Our model of parental care assumes that care decisions are made in the context of an extended family, with each child taking into account not only the parent's needs and the child's own circumstances, but also the characteristics and actual care behavior of siblings. We propose a simultaneous-Tobit statistical framework that embodies these assumptions. The model is estimated using data from the 1993 Asset and Health Dynamics Among the Oldest Old (AHEAD) study. The findings indicate that a child's hours of parent care are reduced, but on much less than a one-for-one basis, as the parent-care hours of siblings increase. We also find that a child's supply of parent-care hours is reduced by having sisters, holding constant the care efforts of siblings. PMID- 9215363 TI - Selection of children to provide care: the effect of earlier parental transfers. AB - We use the first wave of data from the Asset and Health Dynamics Among the Oldest Old (AHEAD) study to examine the effects of past parent-to-child financial transfers on selection of a child to provide assistance with basic personal care for unmarried parents. We estimate a fixed-effects conditional logit model and find a positive and significant association between past financial transfers and a child's current helping behavior. The coefficient of past financial transfers is in the direction hypothesized, and its magnitude is 80% as large as that of gender, a well-documented powerful predictor of parental caregiving. There appears to be substantial evidence that earlier parent-to-child financial gifts play a role in determining which child in the family will provide assistance. PMID- 9215364 TI - Cost-effective evaluation of asymmetric sensorineural hearing loss with focused magnetic resonance imaging. AB - The poor sensitivity of audiometric brain stem response for small vestibular schwannomas (acoustic neuromas) creates a dilemma for the physician evaluating a patient with signs and symptoms of retrocochlear disease. Magnetic resonance imaging is recognized as the gold standard for the evaluation of these problems, but if a complete examination of the internal auditory canals and head is done on every patient, the cost is high. Although less expensive, screening with audiometric brain stem response risks missing up to 33% of small tumors. Therefore we developed a focused magnetic resonance imaging sequence for evaluation of patients with asymmetric sensorineural hearing loss and/or nonpulsatile tinnitus. The protocol includes a T1-weighted sagittal localizer, pregadolinium and postgadolinium T1-weighted 3-mm contiguous axial slices through the internal auditory canal and the region of the cerebellopontine angle, and T2 weighted axial images through the entire brain. Total scanning time is about 12 minutes, and the estimated cost is $300 to $500. We retrospectively reviewed the imaging records of 485 screening examinations done during an 18-month period. Twenty-four patients had diagnoses definitely or probably producing the hearing loss for an overall positive rate of 5%. By eliminating the need for follow-up audiometric or electrophysiologic studies, we believe a focused magnetic resonance imaging-based diagnostic scheme is actually more cost-effective on a cost-perpatient basis. PMID- 9215365 TI - Otolaryngic manifestations in children presenting with apparent life-threatening events. AB - Apparent life-threatening event (ALTE) is a term used to characterize an event of unknown cause after an infant is found limp, cyanotic, bradycardic, and/or requiring resuscitation. Like sudden infant death syndrome (SIDS), ALTE is a general term used until a precise diagnosis can be established. The relationship between ALTE and SIDS has not been clearly defined, although 7 to 15 percent of children with ALTE die of SIDS. If children with ALTE are at greater risk for SIDS, morbidity and mortality may be prevented if the underlying pathology can be identified and corrected or closely monitored. The otolaryngologist is being consulted more frequently to evaluate children who have been through an ALTE to help elucidate any underlying pathology that may have caused the near-death experience. This retrospective chart review reports the evaluation of 30 infants with ALTE requiring consultation by the Division of Pediatric Otolaryngology at the Children's Memorial Hospital in Chicago during a 3-year period. We reviewed the literature and here compare our findings with current animal models. Of the 30 children evaluated, 53% had gastroesophageal reflux, 40% had laryngeal abnormalities, 13% had tracheal abnormalities, and 10% had pharyngeal abnormalities. Thirteen percent of the children had nonotolaryngic anomalies identified during evaluation. Surgical intervention was required in 10 patients and medical treatment was used in 18. When evaluating a child with ALTE, a complete history and physical examination, evaluation for gastroesophageal reflux, assessment for upper airway obstruction by radiographs and endoscopy, and a multidisciplinary approach are recommended. PMID- 9215366 TI - Management of congenital atresia of the external auditory canal. AB - The management of a unilateral congenital atresia of the external auditory canal is nonuniform and debated. Various surgical approaches, timing, coordination with microtia repair, and variable hearing improvements all contribute to the debate regarding management of this entity. This paper outlines our craniofacial team approach to the congenital unilateral atresia and microtia in children. Selection criteria, timing of repair, coordination with microtia repair, surgical results, and pitfalls will be discussed. The results of surgery in 16 patients with unilateral congenital atresia of the external auditory canal and 2 children with bilateral atresia will be presented. Repair of the atresia was undertaken in children 5 years or older who had pneumatized mastoids and middle ears. Replacement of the malleus/incus complex with a partial ossicular reconstruction prosthesis improved closure of the air-bone gap. Drawbacks included meatal stenosis and deepithelization of the split thickness skin graft lining the external auditory canal. Repair of the unilateral congenital atresia is a demanding and challenging problem but one in which excellent results are achievable. PMID- 9215367 TI - Changes in otoacoustic emissions and auditory brain stem response after cis platinum exposure in gerbils. AB - Ototoxicity associated with cis-platinum administration commonly presents as hearing loss and tinnitus. The hearing loss is usually an irreversible, high frequency sensorineural loss. Histologic studies in humans and animals suggest that the outer hair cells (OHCs) are most susceptible to cis-platinum. Evoked otoacoustic emissions (EOAE), as a measure of outer hair cell function, are potentially useful in following ototoxic insults involving OHCs. Distortion product otoacoustic emissions (DPOAE) test frequency-specific regions of the cochlea and therefore may be particularly well suited for monitoring ototoxic injuries. We measured distortion product otoacoustic emissions, at f2 = 2, 4, 6, 8, 10, and 12 kHz, in gerbils after a single large dose of cis-platinum. Animals treated with saline served as controls. The findings were compared to auditory brain stem evoked response (ABR) thresholds, using tone pips of the same frequencies. The DPOAE and ABR thresholds were measured before treatment and again 2, 5, and 14 days after drug administration. The changes in DPOAE were compared with the changes in ABR. No treatment effect was noted in the 2-day group. Animals treated with cis-platinum demonstrated significant elevation of DPOAE and ABR thresholds compared with control animals at 5 and 14 days. There was no significant difference between the threshold changes in the 5- and 14-day groups. PMID- 9215368 TI - Electrocochleography in retrosigmoid vestibular nerve section for intractable vertigo caused by Meniere's disease. AB - Interest in electrocochleography has increased in recent years because of the discovery of an elevated summating potential to action potential amplitude ratio (SP/AP ratio) in patients with endolymphatic hydrops caused by Meniere's disease or perilymph fistula. It was the purpose of this investigation to determine whether the intraoperative SP/AP ratio will decrease after vestibular nerve section in patients with intractable Meniere's disease. Fourteen patients with medically intractable classic Meniere's disease underwent retrosigmoid vestibular nerve section. Intraoperative transtympanic electrocochleography was performed with alternating click stimuli presented at 95 dB HL. In all patients the SP/AP ratio was recorded before the skin incision ("baseline" condition) and after the dura was closed ("closing" condition). Statistical analysis was applied to the recorded data. In 11 (79%) patients, the SP/AP ratio was found to be elevated above 0.30 in the baseline state. In 13 (93%) patients, the SP/AP ratio decreased more than 25% after the nerve was sectioned. These results were highly statistically significant (p < 0.001). We conclude that the SP/AP ratio does decrease in patients with Meniere's disease after undergoing retrosigmoid vestibular nerve section and offer a possible explanation. PMID- 9215369 TI - Neonatal hearing screening with otoscopy, auditory brain stem response, and otoacoustic emissions. AB - A study was performed to investigate the relationship between external and middle ear factors and hearing screening results by auditory brain stem response (ABR) and transient evoked otoacoustic emissions (EOAEs). The ears of 200 well newborns aged 5 hours to 48 hours underwent screening by ABR and EOAEs, followed by otoscopic examination. The pass rates for ABR and EOAE screening were 88.5% and 79%, respectively. On otoscopic examination, 13% (53 of 400) ears had occluding vernix obscuring the view of the tympanic membrane. Cleaning of vernix was attempted in ears that failed ABR or EOAE screening. Seventeen ears that failed ABR were cleaned, and 12 (71%) of them passed repeat ABR. Thirty-three ears that failed EOAE screening were cleaned, and 22 (67%) of them passed repeat emissions testing. Cleaning vernix increased the pass rates for ABR and EOAE screening to 91.5% and 84%, respectively. Decreased tympanic membrane mobility was found in 9% of ears that could be evaluated otoscopically. Increased failure rates for both ABR and EOAE screening were found in infant ears with decreased tympanic membrane mobility, but significance testing could not be performed because of inadequate sample size. Prevalence of occluding external canal vernix and middle ear effusion as a function of increasing infant age were studied. Implications for newborn hearing screening are discussed. PMID- 9215370 TI - Nodular thyroid disease in children and adolescents. AB - Thyroid nodules in children are extremely uncommon. Most thyroid nodules, both benign and malignant, present as asymptomatic neck masses. A thyroid nodule in a child is significant because of the risk of malignancy. A review of medical records at our institution demonstrated 71 patients 20 years of age and younger with surgically managed thyroid nodules, of which 45 were benign and 26 were malignant. Our diagnostic workup, including serum thyroid studies, radiologic evaluation, and fine-needle aspiration, is discussed. Because of the possibility of malignancy, we recommend that all solitary thyroid nodules be excised in children unless fine-needle aspiration definitively determines a benign histology. The extent and type of surgical management is controversial and is still subject to much debate. Partial thyroidectomy appears adequate for benign disease, but even though there is no statistical difference in survival, we recommend total thyroidectomy for the management of malignant disease. PMID- 9215371 TI - Impact of surgical treatment on paranasal fungal infections in bone marrow transplant patients. AB - Invasive fungal sinusitis can develop in immunosuppressed patients. A more complex problem is immunosuppressed patients who have undergone bone marrow transplantation. For a prolonged period, they are both neutropenic and thrombocytopenic. Survival in these patients is poor, and the role for extensive surgical intervention for sinus disease has to be weighed against the risk and the potential that this is a systemic disease. Between January 1983 and June 1993, 29 bone marrow transplant recipients with documented invasive fungal infections of the sinuses and paranasal tissues required surgical intervention. This represents 1.7% of the total 1692 bone marrow transplants performed. There were 22 allogeneic, 6 autologous, and 3 unrelated donor transplants, with two patients receiving two separate grafts. Underlying diseases included 24 hematologic malignancies and 5 other disorders, including 1 aplastic anemia and 1 solid tumor. The mortality rate from the initial fungal infection was 62%. Twenty seven percent resolved the initial infections but subsequently died of other causes. All patients received medical management, such as amphotericin, rifampin, and colony-stimulating factors, in addition to surgical intervention. Surgical management ranged from minimally invasive procedures to extensive resections including medial maxillectomies. Sixty-one percent of the patients who died of the initial infection had undergone extensive surgical procedures versus 55% of those who resolved the infection. Recovery of neutrophil counts was required to clear the infection but did not necessarily predict a good outcome because 50% of those who died of the infection had experienced neutrophil recovery. White blood cell counts at the time of surgery were not significantly different between the two groups. Prognosis was poor when cranial and orbital involvement and/or bony erosion occurred. PMID- 9215372 TI - Extended temporal bone resection for squamous cell carcinoma. AB - OBJECTIVE: The aim of this study was to assess the surgical results of a series of patients from this unit who underwent extended temporal bone resection for recurrent squamous cell carcinoma as a salvage procedure. DESIGN: The surgical records of 15 patients were analyzed in detail. Each patient had salvage surgery in the form of an extended temporal bone resection with supraomohyoid block dissection, dural grafting, and free microvascular forearm or scalp rotation flap repair for recurrent squamous cell carcinoma in a radical mastoid cavity. RESULTS: Radical surgery yielded a 47% 5-year survival. Twenty-nine percent of the survivors had temporal lobe involvement that necessitated a partial excision of the temporal lobe of the brain. Histologic evidence of local lymph node involvement in the supraomohyoid neck dissection was present in 13% of cases. Those who died did so in the first postoperative year. All those with poorly differentiated tumors died. The survivors had well or moderately differentiated tumors. CONCLUSIONS: Radiotherapy alone or partial temporal bone resection, most commonly a radical mastoidectomy with or without preoperative or postoperative radiotherapy is used by the majority of otolaryngologists in treating squamous cell carcinoma of the temporal bone. The 5-year survival rate after this treatment remains depressingly low and the prognosis gloomy, particularly for advanced tumors. The findings in this series of extended temporal bone resections as salvage surgery in recurrent disease is encouraging, and radical surgery combined with radiotherapy from the outset may give much better 5-year survival figures in the future than the conventional partial temporal bone resection and radiotherapy. PMID- 9215373 TI - Trace elements in head and neck cancer patients: zinc status and immunologic functions. AB - In this study we have assessed zinc status and zinc-dependent cell-mediated immune functions (interleukin-2 production by mononuclear cells, natural killer cell lytic activity, and interleukin-1 beta production by mononuclear cells) in adult patients with squamous cell carcinoma of the upper aerodigestive tract at diagnosis and before any therapy was instituted. Inasmuch as significant interactions between zinc, copper, and iron exist, we also assayed the plasma copper level, serum iron level, and total iron-binding capacity in our patients. We recruited 30 cancer subjects and 21 control subjects. On the basis of cellular zinc criteria, we diagnosed a mild deficiency of zinc in 53% of cancer subjects. The plasma zinc level was not decreased in our subjects. A univariate analysis was applied by use of one-way analysis of variance comparing study variables among the three study groups (controls and zinc-deficient and zinc-sufficient cancer patients) and Tukey's multiple comparison test, and we showed that interleukin-2 production and natural killer lytic activity were decreased in zinc deficient cancer patients. Interleukin-1 beta production (ELISA assay) was increased in both zinc-deficient and zinc-sufficient groups. Plasma copper level was not different, but the iron utilization was decreased in both groups of cancer subjects. We conclude that zinc deficiency and zinc-dependent immunologic dysfunctions are present in more than half of the patients with head and neck cancer in the Detroit area. PMID- 9215374 TI - Risk of lung cancer among patients with head and neck cancer. AB - A cohort of 5180 patients with head and neck cancer, who were part of the tumor registry of the Surveillance, Epidemiology, and End Results area of western Washington State, was followed up for as many as 15 years to determine the risk of lung cancer. A sample of 522 patients from this cohort was interviewed to determine smoking history. Lung cancer developed in 356 (6.9%) of the 5180 patients. The overall annual incidence of lung cancer remained relatively constant between approximately 1.0% and 2.0% during the 15 years of follow-up. Men had an increased risk of lung cancer compared with women (relative risk (RR) = 1.56; 95% confidence interval (CI) = 1.18 to 2.03). Compared with patients with oral cavity cancer (RR = 1.00), the relative risk of lung cancer developing by the site of the index tumor was 0.63 (95% CI = 0.40 to 0.98) for lip, 1.12 (95% CI = 0.81 to 1.56) for intrinsic larynx, 1.73 (95% CI = 1.21 to 2.47) for oropharynx, 1.84 (95% CI = 1.16 to 2.92) for hypopharynx, and 2.28 (95% CI = 1.60 to 3.24) for extrinsic larynx. Among the 522 patients who were interviewed, men smoked more than women (p < 0.0001), and patients with laryngeal or pharyngeal cancer smoked more than patients with cancer of the lip or the oral cavity (p < 0.05). Among patients with head and neck cancer, the risk of lung cancer is highest for men and for patients with cancer of the pharynx or extrinsic larynx. These findings may be explained by differences in smoking consumption. PMID- 9215375 TI - Combined surgery and radiation therapy for squamous cell carcinoma of the hypopharynx. AB - Squamous cell carcinoma of the hypopharynx remains a highly lethal disease. This article documents our experience with 132 patients undergoing surgical management of squamous cell carcinoma of the hypopharynx, of whom 80% received postoperative radiation therapy. Local-regional control was obtained in 61% of the patients. Five-year overall and disease-free survival rates were 30% and 41%, respectively. Prognosis was better in patients with limited disease: local disease permitting larynx-sparing surgery, N0/N1 clinical neck, and stage I/II/III disease. Cancer of the hypopharynx remains an aggressive entity associated with poor prognosis. Novel strategies stressing improved local-regional control with prevention of distant metastasis are warranted. PMID- 9215376 TI - Outcome analysis of the transglabellar/subcranial approach for lesions of the anterior cranial fossa: a comparison with the classic craniotomy approach. AB - The classic approach to anterior skull base lesions uses bifrontal craniotomies together with lateral rhinotomies. This approach requires frontal lobe retraction and is associated with postoperative anosmia and the development of frontal lobe encephalomalacia. The transglabellar/subcranial approach permits removal of anterior skull base lesions without frontal lobe retraction and avoids facial scars. No studies to date, however, have directly compared the two approaches in terms of patient morbidity. The present retrospective study compares the two approaches when used for the removal of anterior skull base lesions in terms of estimated blood loss, number of transfusions, number of days in the hospital and intensive care unit, and postoperative complications. Twenty patients with anterior skull base lesions were examined. The classic approach was used on 10, and the transglabellar/subcranial route was used on 10. When compared with the classic approach, the transglabellar/subcranial approach resulted in a lower estimated blood loss and subsequent transfusion rate, fewer days in the hospital and intensive care unit, and lower numbers and less severe types of complications. Furthermore, visualization of the tumors before resection with the transglabellar/subcranial approach allowed preservation of olfaction in virtually all of these patients. Although this study represents a small sample population, the results are sufficiently impressive to favor the transglabellar/subcranial approach for the removal of a variety of anterior skull base lesions. PMID- 9215377 TI - Predictors of residency performance: a follow-up study. AB - In a 1990 study we investigated resident applicant characteristics associated with successful matching into otolaryngology. Of the 175 applicants studied, 87 matched, for a 49.7% success rate. Successful matching was much more likely for applicants with a history of excellent academic achievement in medical school. Of the 88 applicants who did not match during the year that was originally studied, 30 matched to otolaryngology in subsequent years. Of the 58 who never matched in otolaryngology, there is no evidence of board certification for 30. Of the other 28, 12 are board certified in anesthesia; 3 in radiology; 2 each in family medicine, internal medicine, general surgery, psychiatry, and physical medicine, and rehabilitation; and 1 each in pathology, emergency medicine, and dermatology. Of the total of 117 who matched in otolaryngology, 109 began residency training, and 107 finished otolaryngology training. Program directors answered questionnaires about 100 of 107 of these residents, detailing aspects of residency performance. The only correlation found between a highly satisfactory residency performance and characteristics that could be evaluated at the time of interviewing for residency positions was with excellent academic performance in medical school. PMID- 9215378 TI - Endoscopic approach to traumatic visual loss. AB - OBJECTIVE: To review our experience with the use of endoscopic optic nerve decompression in traumatic blindness. METHOD: We did a retrospective analysis of patients with traumatic blindness that underwent endoscopic decompression of the optic canal to determine postoperative visual acuity and correlate it to preoperative visual loss and intraoperative findings. The setting was a Level I university trauma center. We identified 8 patients treated with both surgery and steroids over a 10-month period beginning in 1993 (Seven males, one female). RESULTS: Four of six patients with total blindness (no light perception) had improvements in visual acuity. In three patients, visual acuity returned to preinjury levels. One patients with total blindness was operated on 6 weeks after injury and had a visual acuity of 20/800 at 1-year follow-up. Two patients with hand motion preoperatively had improvement in visual acuity. In one patient, vision returned to normal (20/20), and in the other it improved to 20/200). Five patients were operated on after megadose steroid treatment for at least 48 hours failed; four of five noted dramatic improvements in visual acuity. CONCLUSION: The endoscopic approach may be used to successfully decompress the optic nerve in traumatic blindness. PMID- 9215379 TI - Minimizing upper lip and incisor teeth paresthesias in approaches to transsphenoidal surgery. AB - Currently popular transsphenoidal approaches to the pituitary include sublabial, external rhinoplasty, alotomy, and transnasal techniques. The conventional sublabial approach remains the workhorse method despite postoperative lip edema, potential difficulty for denture wearers, and troublesome persistent upper lip and incisor teeth numbness. We traced the courses of the nasopalatine, infraorbital, and anterior superior alveolar nerves in 41 cadaveric half-head dissection to determine the exact contribution to upper lip and incisor teeth innervation. We then conducted a retrospective patient survey of 25 sublabial, 28 external rhinoplasty, 23 alotomy, and 12 transnasal approaches to the hypophysis to assess the incidence of upper lip and incisor teeth paresthesias lasting longer than 1 month. We conclude that rhinoplastic techniques are superior to the sublabial approach in limiting upper lip and incisor teeth numbness without compromising neurosurgical exposure for hypophysectomy. PMID- 9215380 TI - Problems and pitfalls in community-based outcomes research. AB - OBJECTIVE: To assess the feasibility of multisite community-based outcomes research. DESIGN: Prospective observational study of variations in treating acute external otitis by otolaryngologists and primary care practitioners. SETTING: Community-based independent otolaryngology practices. PARTICIPANTS: Patients with external otitis treated by otolaryngologists in Project Solo, a nonprofit, grassroots organization of independent physicians united for quality, patient advocacy, and cost containment. METHODS: Confidential (bar-coded), disease specific outcomes questionnaires completed by patients (12 items) and by participating physicians (15 items). Response to treatment was measured with a follow-up patient questionnaire (3 items). RESULTS: Nine patients were recruited from 5 of 29 enrolled otolaryngologists. Primary care practitioners were more likely to prescribe oral antimicrobials than otolaryngologists (100% vs. 44%, p = 0.03), but less likely to insert a wick in the external auditory canal (11% vs. 78%, p = 0.02). Poor recruitment was caused by an overly long and complex survey, data collection at multiple time points, lack of time during office hours, cumbersome data collection requirements, inadequate ongoing communication, and a lack of enthusiasm for the project. CONCLUSIONS: Future efforts at implementing a multisite outcomes study will require shorter questionnaires, smoother integration of the survey process with regular office flow, simplified procedures for data exchange, frequent communication with data collection sites, and motivational programs for participating physicians and their office staff. PMID- 9215381 TI - Prospective, longitudinal quality-of-life study of patients with head and neck cancer: a feasibility study including the EORTC QLQ-C30. AB - Despite modern advances in the treatment of head and neck cancer, the survival rate fails to improve. Considering the different treatment modalities involved, quality of life has been thought of as an additional end point criterion for use in clinical trials. A Nordic protocol to measure the quality of life of head and neck cancer patients before, during, and after treatment was established. Before the study, a pilot study was done with this protocol. The main purpose of this pilot study was to find out whether this cancer population would answer quality of-life questionnaires repeatedly (six times) over a 1-year period and whether the chosen questionnaires-a core questionnaire (European Organisation for Research and Treatment of Cancer Core Quality of Life Questionnaire (EORTC QLQ C30)), a tumor-specific questionnaire, and a psychological distress measure (Hospital Anxiety and Depression scale (HAD))-were sensitive for changes to functions and symptoms during the study year. The results presented in this article all refer to the pilot study. Forty-eight consecutive patients agreed to participate in the study. The most common tumor locations were the oral cavity (17) and the larynx (12). Almost all patients received combined treatment: 45 of 48 radiation therapy, 18 of 48 chemotherapy, and 17 of 48 surgery. After the primary treatment, 40 patients had complete tumor remission. Four of the 48 patients did not answer any questionnaires and were therefore excluded from the study. Of the remaining 44 patients, 3 died during the study year, and another 6 withdrew for various reasons. Thirty-five (85%) of the 41 patients alive at the 1 year follow-up answered all six questionnaires and thus completed the study. Mailed questionnaires were used throughout the study. All questionnaires were well accepted and found to be sensitive to changes during the study year. The greatest variability was found for symptoms and functions related specifically to head and neck cancer. The symptoms were swallowing difficulties, hoarse voice, sore mouth, dry mouth, and problems with taste. They all showed the same pattern, with an increase of symptoms during and just after finishing the treatment. The HAD scale revealed a high level of psychological distress, with 21% probable cases of psychiatric morbidity at diagnosis. In conclusion, it was shown that the study design and questionnaires were feasible for the forthcoming prospective quality-of-life assessment of Swedish and Norwegian head and neck cancer patients. PMID- 9215382 TI - Primary peripheral T-cell lymphoma of the acoustic nerve. PMID- 9215383 TI - Giant cholesteatoma presenting as a postauricular mass. PMID- 9215385 TI - Metastatic malignant mesothelioma to the tonsil. PMID- 9215384 TI - Phrenic nerve paralysis after doxycycline sclerotherapy for chylous fistula. PMID- 9215386 TI - New laser ruler instrument for making measurements through an endoscope. PMID- 9215387 TI - Technique for promoting healing of complex tracheostomy wounds. PMID- 9215388 TI - Use of the miniarthroscopic drill in choanal atresia repair: how we do it. PMID- 9215389 TI - Use of steerable forceps in endoscopic sinus surgery. PMID- 9215390 TI - Amyloidosis. PMID- 9215391 TI - Superior thyroid artery arising from the common carotid artery. PMID- 9215392 TI - Stoma recurrence after laryngectomy. PMID- 9215393 TI - Hyperbaric oxygen therapy. PMID- 9215394 TI - Effect of active immunization against luteinizing hormone-releasing hormone on the androgen-sensitive Dunning R3327-PAP and androgen-independent Dunning R3327 AT2.1 prostate cancer sublines. AB - BACKGROUND: The objective of this study was to determine the effect of active immunization against LHRH on the growth characteristics and histology of subcutaneously implanted tumors of the androgen-sensitive Dunning R3327-PAP and androgen-independent R3327-AT2.1 rat prostate adenocarcinoma sublines. RESULTS: We herein demonstrate that 1) active immunization with an LHRH-diphtheria toxoid conjugate (LHRH-DT) leads to the downregulation of gonadotropins and testosterone and consequently the atrophy of testosterone-dependent organs such as the testes, prostate, and androgen-sensitive Dunning R3327-PAP tumors, 2) growth inhibition of Dunning R3327-PAP tumors is caused by suppression of cell division rather than by an increase in cell death and is associated with an increase of the tumor stroma content, and 3) volume increase of the androgen-independent Dunning R3327 AT2.1 tumor is slightly but significantly reduced, indicating a local stimulatory LHRH loop within this tumor cell line. PMID- 9215395 TI - Ischemic damage to the prostate during cardiac surgery: a clinical model. AB - BACKGROUND: To determine if altered tissue perfusion during cardiac surgery results in ischemic tissue damage to the prostate, as suggested by a rise in prostatic-specific antigen (PSA). METHODS: Twenty-nine male patients undergoing elective coronary artery bypass grafting were studied. Ten male patients undergoing elective gastrointestinal surgery served as controls. PSA levels were determined preoperatively and six hourly intervals postoperatively for 48 hr. All patients underwent urethral catheterization at induction of anesthesia. RESULTS: All patients (100%) who had undergone cardiac bypass surgery showed rises in serum PSA during 48 hr of postoperative follow-up. At the 6-hr postoperative interval, the mean PSA was significantly different from the mean baseline value (paired two tailed Student's t test, P < 0.001) in 27 of the 29 (93%) patients. In contrast, the PSA values in the 10 gastroenterological controls did not change at 6 hr (P > 0.2) or during the next 48 hr. One patient in the cardiac group showed a very marked elevation in serum PSA of greater than 50 times normal preoperative levels. CONCLUSIONS: Statistically significant rises in PSA levels are seen following coronary bypass surgery. This rise may be caused by ischemic nontrauma related damage to the prostate and suggests a possible pathophysiological mechanism for the clinically episodic symptoms of prostatism seen in elderly men. PMID- 9215396 TI - Interstitial photodynamic therapy in the canine prostate with disulfonated aluminum phthalocyanine and 5-aminolevulinic acid-induced protoporphyrin IX. AB - BACKGROUND: Photodynamic therapy (PDT) is an experimental approach for treating prostate cancer localized to the gland that does not involve surgery or irradiation. Second-generation photosensitizers 5-aminolevulinic acid (ALA) and aluminum disulfonated phthalocyanine (AlS2Pc) were studied in the normal canine prostate. METHODS: Tissue biodistribution of photosensitizers on serial biopsies was examined using fluorescence microscopy. Photodynamic therapy was done by delivering red light interstitially at 100 mW through fibers placed under transrectal ultrasound guidance. RESULTS: Peak levels of AlS2Pc appeared at 5-24 hr and at 3 hr for ALA. Macroscopic PDT lesions were up to 12 mm in diameter using AlS2Pc, but only 1-2 mm with ALA. Light at 300 mW caused thermal lesions. At 28 days, damaged glands remained atrophic, but the interlobular supporting stroma was well-preserved. Urethral lesions healed by 28 days without functional impairment. CONCLUSIONS: Although the results with ALA were disappointing, PDT using AlS2Pc looks like a promising modality for treatment of localized prostate cancer. PMID- 9215397 TI - Expression of p21 and mutant p53 gene products in residual prostatic tumor cells after radical radiotherapy. AB - BACKGROUND: In a previous study, sextant core biopsies revealed residual tumor in the prostate in 37/55 investigated patients, with an average of 6.8 years after external beam radiation therapy (RRT). More than half of the biopsies exhibited Ki-67 and PCNA proliferation activity. METHODS: The present study aims at further characterizing residual tumor cells post-RRT by investigating whether the tumor cells exhibit immunohistochemical expression of p21 and mutant p53 gene products, which reflect the state of cell cycle regulatory mechanisms. RESULTS: Positive p53 staining was observed in 11% and p21 positivity in 47% of biopsies. The proportion of positively stained cells was low for both antigens. The staining patterns point to the existence of wild-type p53-dependent, as well as alternative pathways for p21 protein induction. CONCLUSIONS: A low proportion of tumor cells exhibited p53 protein accumulation post-RRT. G1 arrest, as assessed by p21 immunoexpression, was demonstrated in a low percentage of tumor cells in < 50% of post-RRT biopsies, indicating that the vast majority of residual tumor cells following RRT escape the G1/S checkpoint control and propagate into S phase, presumably with a maintained malignant potential. PMID- 9215398 TI - Synergistic activation of androgen receptor by androgen and luteinizing hormone releasing hormone in prostatic carcinoma cells. AB - BACKGROUND: We investigated modulation of androgen receptor (AR) activity in prostatic tumor cells by luteinizing hormone-releasing hormone (LHRH)-induced increase of the intracellular cyclic adenosine monophosphate (cAMP) level. METHODS: AR transactivation activity was assessed in transiently transfected DU 145 and in LNCaP cells. RESULTS: LHRH and cAMP derivative, respectively, induced reporter gene activity to about 15% of the maximal level in DU-145 cells transfected with an AR expression vector and an androgen-inducible reporter gene. LHRH or the cAMP analogue acted synergistically in combination with low concentrations of androgen thus lowering the androgen concentration required for maximal AR activation by a factor of 100. A similar activation of the AR by cAMP analogue was observed in LNCaP cells when enhancement of androgen-induced secretion of prostate-specific antigen was determined. The two nonsteroidal antiandrogens hydroxyflutamide and Casodex(R) inhibited reporter gene activity. CONCLUSIONS: The AR is synergistically activated by low doses of androgen and LHRH or the second messenger cAMP. This may have implications for the treatment of advanced prostate cancer. PMID- 9215399 TI - Effects of retinoid X receptor-selective ligands on proliferation of prostate cancer cells. AB - BACKGROUND: Management of prostate cancer that either is detectable by prostate specific antigen (PSA) measurements after curative intent or has spread outside of its capsule is a serious problem. Innovative, nontoxic approaches to the disease are required. One approach might be therapy with retinoids. Retinoid activities are mediated by two distinct families of transcription factors: the retinoic acid receptors (RARs) and retinoid X receptors (RXRs), which can induce transcriptional activation through specific DNA sites or by inhibiting the transcription factor AP-1 that usually mediates cellular proliferative signals. The RARs require heterodimerization with RXRs. RXRs can form either heterodimers or homodimers; and the latter can bind to DNA response elements that are distinct from those bound by the RAR/RXR heterodimers. METHODS: A series of novel synthetic retinoids that selectively interact with RXR/RXR homodimers or RAR/RXR heterodimers, or that selectively inhibit AP-1 activity without activating transcription were evaluated for their ability to inhibit clonal growth of three human prostate cancer cell lines (PC-3, DU-145, and LNCaP). RESULTS: Several notable findings were: 1) RXR-selective retinoids, such as SR11246, were able to inhibit the clonal growth of prostate cancer cells. In contrast, SR11246 had little effect on clonal growth of myeloid leukemic cells. 2) RAR-selective retinoids also inhibited clonal growth of prostate cancer cells. 3) The retinoid (SR11238) with potent anti-AP-1 activity had no effect on the clonal growth of prostate cancer cells. CONCLUSIONS: This study shows that both RXR- and RAR selective retinoids are worthy of further study and may be candidates for future clinical trials in prostate cancer. PMID- 9215400 TI - Lignans and isoflavonoids in plasma and prostatic fluid in men: samples from Portugal, Hong Kong, and the United Kingdom. AB - BACKGROUND: Chinese men have lower incidences of prostate cancer compared to men from Europe and North America. Asians consume large quantities of soya, a rich source of isoflavanoids phyto-oestrogens and have high plasma and urinary levels of these compounds. The mammalian lignans, enterolactone and enterodiol, are another group of weak plant oestrogens and are derived from seeds, cereals and grains. Vegetarians have high plasma and urinary concentrations of lignans. METHODS: The concentrations lignans and isoflavonic phyto-oestrogens were determined by gas chromatography-mass spectrometry (GC-MS) in plasma and prostatic fluid from Portuguese, Chinese and British men consuming their traditional diets. RESULTS: In prostatic fluid the mean concentrations of enterolactone were 31, 162 and 20.3 ng/ml for Hong Kong, Portugal and Britain respectively. Very high levels of enterolactone (> 600 ng/ml) were observed in the prostatic fluid of some of the men from Portugal. High concentrations of equol (3270 ng/ml) and daidzein (532 ng/ml) were found in a sample of prostatic fluid from Hong Kong. Higher mean levels of daidzein were observed in prostatic fluid from Hong Kong at 70 ng/ml, compared to 4.6 and 11.3 ng/ml in samples from Portugal and Britain respectively. Mean levels of daidzein were higher in the plasma samples from Hong Kong (31.3 ng/ml) compared to those from Portugal (1.3 ng/ml) and Britain (8.2 ng/ml). In general, the mean plasma concentrations of enterolactone from the three centres were similar, at 6.2, 3.9 and 3.9 ng/ml in samples from Hong Kong Portugal and Britain respectively. CONCLUSIONS: Higher concentrations of the isoflavanoid phyto-oestrogens, daidzein and equol, were found in the plasma and prostatic fluid of men from Hong Kong compared to those from Britain and Portugal. However, the levels of the lignan, enterolactone, were very much higher in prostatic fluid of Portuguese men. Isoflavanoids and lignans have many interesting properties and may, in part, be responsible for lower incidences of prostate cancer in men from Asia and also some Mediterranean countries. The isoflavanoids from soya, which are present in high concentrations in the prostatic fluid of Asian men, may be protective against prostate disease. PMID- 9215401 TI - Large fragment of the probasin promoter targets high levels of transgene expression to the prostate of transgenic mice. AB - BACKGROUND: Androgen regulation and prostate-specific expression of targeted genes in transgenic mice can be controlled by a small DNA fragment of the probasin (PB) promoter (-426 to +28 base pairs, bp). Although the small PB fragment was sufficient to direct prostate-specific expression, the low levels of transgene expression suggested that important upstream regulatory sequences were missing. METHODS: To enhance transgene expression, a large fragment of the PB promoter (LPB, -11,500 to +28 bp) was isolated, linked to the bacterial chloramphenicol acetyl transferase (CAT) gene, and microinjected into CD1 mouse oocytes to generate transgenic mouse lines. RESULTS: As shown by the immunohistochemical studies, CAT gene expression was restricted to the prostatic epithelial cells in a tissue-specific manner. High levels of CAT gene expression were observed in two of the six LPB-CAT transgenic lines. In Line 1, developmental regulation of LPB-CAT was detected early, from 1 to 4 weeks of age, with the activity of CAT increasing from 3 to 40,936 dpm/min/mg protein. Upon sexual maturation and elevated serum androgen levels (7 weeks of age), a further 18-fold rise in CAT activity occurred. Hormone ablation by castration in mature mice dramatically reduced transgene expression, whereas treatment with androgens returned LPB-CAT expression to precastration levels. In contrast, treatment with glucocorticoids had no significant effect on CAT gene expression. Zinc treatment of the castrated animals also increased LPB-CAT expression three- to four-fold in two prostatic lobes. CONCLUSIONS: This study demonstrates that important regulatory DNA sequences located in the LPB fragment contribute to tissue specific expression and greatly increase levels of transgene expression induced by androgens and zinc. PMID- 9215402 TI - Prostate-specific membrane antigen. AB - BACKGROUND: In an effort to discover new prostate-specific antigens (PSAs) to enhance our understanding of the functions and behavior of the prostate and the complex processes involved in prostate tumor progression, the structure and function of the PSM antigen has been elucidated. METHODS: The PSM antigen was recognized using the 7E11-C5.3 monoclonal antibody, generated against the LNCaP human prostate adenocarcinoma cell line. The PSM cDNA was isolated by PCR, using tryptic peptides of immunoprecipitated PSM to design degenerate primers. RESULTS: The prostate specific membrane antigen (PSM) is a 100 KD glycoprotein which appears to be a type II integral membrane protein. The protein and cDNA have been extensively characterized and the findings reviewed in the report. CONCLUSIONS: PSM, a new prostate antigen is valuable as a marker for hematogenous micro metastatic tumor dissemination as detected in RT-PCR assays of peripheral blood. PSM has many properties that may be potentially useful as a molecular target in monoclonal antibody directed strategies of tumor imaging and therapy. PMID- 9215403 TI - Origins of radical perineal and nerve-sparing retropubic prostatectomy. PMID- 9215404 TI - The issue of safe testing for brucellosis in Yellowstone bison. PMID- 9215405 TI - The myth of stolen-animal trafficking. PMID- 9215406 TI - In support of changes in veterinary curricula. PMID- 9215407 TI - What is your diagnosis? Herniation of the small intestine through the dorsal lumbar musculature, multiple pelvic fractures, sacroiliac subluxation, and a right craniodorsal coxofemoral luxation. PMID- 9215408 TI - Suppression of gastric acidity in horses. PMID- 9215409 TI - Extralabel use of oxytetracycline. PMID- 9215410 TI - The statutory offense of cruelty to animals. PMID- 9215411 TI - Comparison of results of hormonal analysis of samples obtained from selected venous sites versus cervical ultrasonography for localizing parathyroid masses in dogs. AB - OBJECTIVE: To compare a technique in which samples obtained from selected venous sites are analyzed for parathyroid hormone (PTH) concentration versus usefulness of cervical ultrasonography for localizing primary hyperparathyroidism (PHP) in dogs. DESIGN: Prospective study. ANIMALS: 12 dogs with PHP. PROCEDURE: For each dog, blood samples were collected from the left and right jugular veins and 1 cephalic vein for determination of serum PTH concentration. Ultrasonography of the neck was performed in each dog. Each dog underwent exploratory surgery of the neck. Abnormal appearing parathyroid tissue was removed. Dogs were included in the study if serum calcium concentration decreased within 12 hours after surgery, hypercalcemia completely resolved within 96 hours after surgery, and serum calcium concentration was maintained within the reference range for at least 6 months after surgery. RESULTS: Serum PTH concentrations from the 3 veins were similar in 11 of 12 dogs with PHP. In 1 dog, the serum PTH concentration from the jugular vein ipsilateral to a parathyroid adenoma was greater than that from the contralateral jugular or cephalic vein. Ultrasonography correctly identified a parathyroid mass and its location in 10 of 11 dogs with a solitary abnormal parathyroid gland and in 1 dog in which both parathyroid glands were enlarged. CLINICAL IMPLICATIONS: Surgeons may benefit from knowing the location of abnormal parathyroid tissue in dogs with PHP before surgical exploration. Ultrasonography has potential value for identifying and localizing abnormal parathyroid tissue, whereas sample collection from selected sites for PTH analysis is not likely to be helpful. PMID- 9215412 TI - Short-term results and complications of prepubertal gonadectomy in cats and dogs. AB - OBJECTIVE: To determine short-term results and complications of prepubertal gonadectomy in cats and dogs. DESIGN: Prospective randomized study. ANIMALS: 775 cats and 1,213 dogs. PROCEDURE: Animals undergoing gonadectomy were allotted into 3 groups on the basis of estimated age (group 1, < 12 weeks old; group 2, 12 to 23 weeks old; group 3, > or = 24 weeks old). Complications during anesthesia, surgery, and the immediate postoperative period (7 days) were recorded. Complications were classified as major (required treatment and resulted in an increase in morbidity or mortality) or minor (required little or no treatment and caused a minimal increase in morbidity). An ANOVA was used to detect differences among groups in age, weight, body temperature, and duration of surgery. To detect differences in complication rates among groups, chi 2 analysis was used. RESULTS: Group 1 consisted of 723 animals, group 2 consisted of 532, and group 3 consisted of 733. Group-3 animals had a significantly higher overall complication rate (10.8%) than group-1 animals (6.5%), but did not differ from group-2 animals (8.8%). Differences were not detected among the 3 groups regarding major complications (2.9, 3.2, and 3.0% for groups 1, 2, and 3, respectively), but group-3 animals had significantly more minor complications (7.8%) than group-1 animals (3.6%), but not group-2 animals (5.6%). CLINICAL IMPLICATIONS: In this study, prepubertal gonadectomy did not increase morbidity or mortality on a short term basis, compared with gonadectomy performed on animals at the traditional age. These procedures may be performed safely in prepubertal animals, provided that appropriate attention is given to anesthetic and surgical techniques. PMID- 9215414 TI - Surgical removal of heartworms from the right atrium of a cat. AB - As few as 2 adult heartworms can cause cardiac enlargement and severe respiratory compromise in cats. Use of echocardiography to view heartworms may indicate whether surgical removal is possible. Aided by fluoroscopy, manual insertion of a catheter that contains basket-type retrieval forceps into the right atrium, via jugular venotomy, may permit removal of adult heartworms from cats. Surgical removal of heartworms and supportive treatment with heparin, prednisone, and antibiotics may result in complete recovery from heartworm infection in cats. PMID- 9215413 TI - Ipodate treatment of hyperthyroidism in cats. AB - OBJECTIVE: To evaluate the efficacy and safety of ipodate in the treatment of hyperthyroidism in cats. DESIGN: Prospective case series. ANIMALS: 12 cats with hyperthyroidism treated at The Animal Medical Center between November 1994 and March 1996. PROCEDURE: Each cat initially received 100 mg of ipodate/d, PO. The drug's effects on clinical signs, body weight, heart rate, and serum triiodothyronine (T3) and thyroxine concentrations were evaluated 2, 4, 6, 10, and 14 weeks after initiation of treatment. A CBC and serum biochemical analyses were performed at each evaluation to monitor potential adverse effects of the drug. Dosage of ipodate was increased to 150 mg/d and then to 200 mg/d at 2-week intervals if a good clinical response was not observed. RESULTS: 8 cats responded to treatment and 4 did not. Among cats that responded, mean body weight increased and mean heart rate and serum T3 concentration decreased during the study period. Among cats that did not respond, mean body weight decreased and mean heart rate and serum T3 concentration were not significantly changed. Serum thyroxine concentration remained high in all cats. Adverse clinical signs or hematologic abnormalities attributable to ipodate treatment were not reported in any of the cats. CLINICAL IMPLICATIONS: Ipodate may be a feasible alternative to methimazole for medical treatment of hyperthyroidism in cats, particularly those that cannot tolerate methimazole and are not candidates for surgery or radiotherapy. Cats with severe hyperthyroidism are less likely to respond to ipodate than are cats with mild or moderate disease, and cats in which serum T3 concentration does not return to the reference range are unlikely to have an adequate improvement in clinical signs. PMID- 9215415 TI - Suspected microscopic hepatic arteriovenous fistulae in a young dog. AB - Portal hypertension can develop from any disorder that obstructs portal blood flow and may cause ascites in young dogs. Anomalous hepatic arteriovenous (AV) connections are rare but should be suspected in any young dog with portal hypertension or ascites. All previous reports of dogs with hepatic AV fistulae have documented macroscopic connections between the arterial and venous systems. Identical clinical signs and histopathologic findings can develop in dogs in which a macroscopic hepatic AV connection cannot be detected. Microscopic AV connections may be responsible for clinical signs in these dogs. PMID- 9215416 TI - Risk factors associated with development of pelvic canal stenosis secondary to sacroiliac separation: 84 cases (1985-1995). AB - OBJECTIVE: To measure pelvic canal diameter in dogs from a ventrodorsal radiographic view of the pelvic region, to define a normal pelvic canal diameter, to evaluate risk factors associated with stenosis of the pelvic canal secondary to sacroiliac separation, and to determine clinical signs associated with pelvic canal stenosis. DESIGN: Retrospective case series. ANIMALS: 84 case-group and 46 control-group dogs. PROCEDURE: Medical records and radiographs of dogs with conditions unrelated to pelvic fracture (control group) and dogs with sacroiliac separation (case group) in which radiographs were obtained before surgery, after surgery, or after fracture healing were reviewed. Discriminant analysis was used to determine a normal pelvic canal diameter. An ANOVA and Dunnett's two-sided test were used to determine factors associated with pelvic canal stenosis. RESULTS: Pelvic canal diameter ratio determined from control-group dogs was > or = 1.1. Pelvic canal diameter ratios were significantly less for case-group dogs on radiographs obtained before surgery and after fracture healing than for control-group dogs, regardless of fracture type or treatment, except for dogs with ilial fractures treated conservatively. Pelvic canal diameter ratios did not differ for case-group dogs on radiographs obtained after surgery from those for control-group dogs, except when ilial fractures were surgically reduced. None of the dogs had clinical signs associated with pelvic canal stenosis. CLINICAL IMPLICATIONS: Pelvic canal diameter in dogs can be determined from a ventrodorsal radiographic view of the pelvic region. Dogs with pelvic fractures that have a normal pelvic canal diameter before surgery tend to have a normal pelvic canal diameter after fracture healing. PMID- 9215417 TI - Results of a combined dexamethasone suppression/thyrotropin-releasing hormone stimulation test in healthy horses and horses suspected to have a pars intermedia pituitary adenoma. AB - OBJECTIVE: To evaluate results of a combined dexamethasone suppression/thyrotropin-releasing hormone (TRH) stimulation test in horses suspected clinically to have a pars intermedia pituitary adenoma (PIPA). DESIGN: Case-control study. ANIMALS: 7 healthy adult horses and 5 horses suspected to have a PIPA. PROCEDURE: A baseline blood sample was collected, and dexamethasone (40 micrograms/kg [18 micrograms/lb] of body weight, IV) was administered; a second blood sample was collected 3 hours later, and TRH (1.1 mg, IV) was administered; serial blood samples were collected 15, 30, 45, 60, and 90 minutes and 21 hours after TRH administration (24 hours after dexamethasone injection). Cortisol concentration was determined for all blood samples. RESULTS: Baseline cortisol concentration was significantly lower in horses suspected to have a PIPA than in healthy horses. Cortisol concentration was suppressed by dexamethasone in both groups; however, after TRH administration, cortisol concentration returned to baseline values in horses suspected to have a PIPA, but not in healthy horses. Concentration was still less than the baseline value 24 hours after dexamethasone administration in healthy horses. CLINICAL IMPLICATIONS: The combined dexamethasone suppression/TRH stimulation test may be a useful diagnostic test in horses suspected to have a PIPA. For clinical application, collection of a blood sample 30 minutes after TRH administration is recommended. PMID- 9215418 TI - Magnesium toxicosis in two horses. AB - Magnesium sulfate, a saline laxative, is often used for treatment of intestinal impactions in horses. Clinical signs of hypermagnesemia are an uncommon complication following oral administration of magnesium sulfate. Overdose of magnesium sulfate in combination with renal insufficiency, hypocalcemia, or compromise of intestinal integrity may predispose horses to magnesium toxicosis. Establishment of diuresis with fluids and IV administration of calcium may provide successful treatment of magnesium toxicosis in horses. PMID- 9215419 TI - Multiple abortions associated with caprine herpesvirus infection in a goat herd. AB - Herpesvirus infection of pregnant goats may result in abortion of fetuses without other signs of infection in does. Fetuses aborted as a result of caprine herpesvirus infection have tissue necrosis and intranuclear inclusion bodies in multiple organs. PMID- 9215420 TI - Making a profession visible: three nurses' stories. PMID- 9215421 TI - Stitch, stitch ... creating an effective pain management program for critically ill patients. PMID- 9215422 TI - Health practices of critical care nurses: are these nurses good role models for patients? AB - BACKGROUND: Few studies have explored the health practices of critical care nurses. Critical care nurses routinely teach patients about using healthy practices such as low-fat diets, exercise, and routine screening examinations. However, it may be even more important that the nurses themselves have a healthy lifestyle, thus serving as role models for patients. Nurses are selling a product, and that product is health. The best salespersons are those who are genuinely committed to their product and model its benefits. Therefore, critical care nurses' healthful practices can have a profound effect on their patients. OBJECTIVES: The purpose of this descriptive exploratory study was to examine critical care nurses' responses to three questions about health practices in their daily lives: (1) What are critical care nurses doing currently to stay healthy? (2) Do they anticipate making any changes in their lifestyle in the future? (3) Would they recommend their lifestyle to their patients? METHODS: One hundred twenty-seven critical care nurses attending a midwestern critical care conference completed a two-part questionnaire designed to produce a health profile. In a man-on-the-street approach, 23 nurses participated in an interview via video camera. Descriptive statistics were used to analyze the data retrieved from the questionnaires. Interviews were transcribed verbatim and analyzed for themes with a constant comparative method. RESULTS: More than 70% of the critical care nurses who responded engage in exercise and follow a healthy, low-fat diet. Seventy-one percent said that they anticipate making a change in their lifestyle in the future, and 70% said that they would recommend their lifestyle to their patients. Five themes emerged from the videotaped interviews: (1) Heart-healthy practices predominated the responses. (2) Incorporating a healthy lifestyle was easy for some and a struggle for others. (3) Critical care nurses readily listed barriers to healthy living. (4) The nurses had a positive attitude about their healthy lifestyles and felt optimistic about being role models for their patients. (5) Future plans were either singular in focus or limited to maintenance of current health habits. CONCLUSIONS: The majority of the nurses reported practicing a healthy lifestyle and thought that they were good role models for patients. PMID- 9215423 TI - Recognizing a heart attack: the process of determining illness. AB - OBJECTIVE: To examine the process by which patients with acute myocardial infarction recognize illness and the need for medical treatment. DESIGN: Descriptive, exploratory, qualitative. SETTING: The coronary care and progressive care units of two midwestern medical centers. SUBJECTS: Thirty men and women with a diagnosis of acute myocardial infarction. METHODS: Open-ended interviews were conducted on the fourth or fifth day of hospitalization. All interviews were recorded on audiotape and transcribed. Data were analyzed by using grounded theory methods. RESULTS: Findings of the study indicated that determining illness and the need for medical attention was often a difficult process involving phases. The first phase involved attending to or ignoring bodily sensations as they come and go or additional sensations develop. Some subjects moved precipitously to seeking treatment; others took days to attend to bodily sensations. The second phase involved comparing sensations with those from a previously experienced illness or with the subject's concept of sensations likely to accompany common ailments such as indigestion or flu or more serious illnesses such as ulcer, gallbladder disease, or heart attack. The quality of sensations experienced had an important influence on assigning probable cause and deciding that medical attention is warranted. CONCLUSIONS: The disruptive nature of signs and symptoms and how closely signs and symptoms matched the subject's prototype for a heart attack greatly influenced the determination that illness was present and healthcare was needed. These findings have implications for educating the public about the complex and variable manifestations of a heart attack. PMID- 9215424 TI - Managing patients with primary pulmonary hypertension: prostacyclin therapy. PMID- 9215425 TI - Nursing care of patients undergoing thoracoscopic minimally invasive bypass grafting. AB - Recent efforts in cardiac surgery to decrease cost, length of hospitalization, and morbidity without compromising care or outcomes include a method of treatment of coronary heart disease called minimally invasive coronary artery bypass grafting. This article describes the use of thoracoscopy without sternotomy, cardiopulmonary bypass, or cardioplegic arrest for grafting of the left anterior descending branch of the coronary artery with the left internal mammary artery. Preliminary information supports the concept that thoracoscopic minimally invasive bypass grafting is a less costly and less invasive procedure for patients with disease of the left anterior descending branch of the coronary artery. PMID- 9215427 TI - Effect of a monitored care unit on resource utilization in a pediatric ICU. AB - OBJECTIVE: To determine the effect of a monitored care unit on resource utilization in a pediatric ICU. METHODS: The study was done at a 205-bed pediatric medical center located in northern California. Efficiency of resource utilization in the pediatric ICU was evaluated by comparing the following factors before and after the implementation of a monitored care unit: (1) efficiency in the pediatric ICU, (2) number of patients turned away for lack of beds, (3) hours of nursing care per patient day, and (4) cost (as estimated from charge ratios) in the monitored care unit for care of patients admitted because of some common reasons. RESULTS: Efficiency in the pediatric ICU increased significantly after implementation of a monitored care unit. The number of patients not admitted to the pediatric ICU because not enough beds were available was identical before and after establishment of the monitored care unit. After the monitored care unit was established, hours of nursing care per patient day were significantly reduced, enabling the number of patient days to be increased without increasing cost. In addition, cost comparisons showed a decrease in both length of stay and total cost per admission for patients undergoing ventriculoperitoneal shunt revision and lambdoidal suture synostectomy. CONCLUSIONS: Use of beds in the pediatric ICU was more efficient when a high-observation setting was available for low-risk monitored patients. Key differences in patterns of use were observed. Compared with the pediatric ICU, the monitored care unit requires fewer personnel and less expensive equipment and supplies, but it still allows potentially life threatening complications to be recognized and treated. For patients who meet its admission criteria, the monitored care unit is a safe alternative to the pediatric ICU. PMID- 9215426 TI - A survey of pediatric critical care nurses' knowledge of pain management. AB - BACKGROUND: Although nurses are accountable for pain management, it cannot be assumed that they are well informed about pain. Nurses' knowledge base underlies their pain management; therefore, it is important to measure their knowledge. OBJECTIVE: To measure pediatric critical care nurses' knowledge of pain management. METHOD: A descriptive, exploratory study was done. After a pilot study, an investigator-developed Pain Management Knowledge Test was distributed to 50 pediatric ICU nurses. Test responses were collected anonymously and coded by number. Item analysis was done, and descriptive statistics were calculated. Modified content analysis was used on requests for pain-related information. RESULTS: The test return rate was 38%. The overall mean score was 63%. Mean scores within test subsections varied from 50% to 92%. Other mean scores were 85% on a nine-item scale of drug-action items and 92% on a two-item scale of intervention items. However, no nurse recognized that cognitive-behavioral techniques can inhibit transmission of pain impulses; only 32% indicated that meperidine converts to a toxic metabolite, only 47% recognized nalbuphine as a drug that may cause signs and symptoms of withdrawal if given to a patient who has been receiving an opioid; and only 63% indicated that when a child states that the child has pain, pain exists. Thirteen nurses requested pain-related information, and all requests focused on analgesic medications. CONCLUSIONS: Testing nurses' knowledge of pain indicated gaps that can be addressed through educational interventions. Research is needed in which the test developed for this study is used as both pretest and posttest in an intervention study with pediatric critical care nurses or is modified for use with nurses in other clinical areas. PMID- 9215428 TI - Care and safety of pacemaker electrodes in intensive care and telemetry nursing units. AB - BACKGROUND: Temporary pacing leads with electrodes are a potential risk, because microshock inadvertently transmitted across the catheter or wire can paradoxically cause lethal dysrhythmias. Much attention has been paid to this complication in clinical guidelines, but little is known about actual practices used to protect patients. OBJECTIVES: A national survey was done to describe current practices related to the care of patients with temporary epicardial or transvenous pacing catheters. The survey focused on environmental factors that affect generation of static electricity, equipment used with temporary pacing, and nursing practices used when handling temporary pacing electrodes. METHODS: The Pacemaker Electrical Care and Safety Survey was developed, validated, and pilot tested before it was mailed to all 895 hospitals that perform cardiac surgery. Surveys were sent to the coronary care unit, cardiac surgical ICU, and telemetry units of each hospital. RESULTS: Responses were received from 476 units representing 388 (43%) of the 895 institutions. Most respondents reported using gloves, although few hospitals had policies mandating this practice. The insulating materials used most often, in order, were a glove or finger cot, tape, and gauze. Few units (25%) use any measure to reduce static electricity generated by movement over carpeting. Little attention was paid to insulating exposed epicardial temporary pacing electrodes at the generator. CONCLUSIONS: Temporary pacing electrodes were usually handled in an electrically safe manner; however, little attention was paid to environmental sources of microshock or connections between the generator and the cable. Although the respondents reported using a variety of insulating materials, the ideal cover for the exposed tips of the electrodes has not yet been determined. PMID- 9215429 TI - Accuracy of detection of clinically important dysrhythmias with and without a dedicated monitor watcher. AB - BACKGROUND: Although dedicated monitor watchers are used in many progressive care units with telemetry monitors, this costly practice has not been evaluated. OBJECTIVE: To compare the accuracy of detection of clinically important dysrhythmias with and without a dedicated monitor watcher. METHODS: On a 26-bed cardiac progressive care unit, documentation of four categories of dysrhythmias during a 7-week period when a monitor watcher was present was compared with that during a 7-week period when no monitor watcher was present. The Hewlett-Packard CareVue Clinical Event Review, a full-disclosure system, was used as the gold standard. RESULTS: Accuracy of detection of nonsustained ventricular tachycardia, supraventricular tachycardia, and pauses was significantly better with a monitor watcher than without. Although the detection of life-threatening rhythms was correct a higher percentage of the time with a monitor watcher, the difference was not significant. CONCLUSIONS: The efficiency and quality of patient care can be enhanced by using a dedicated monitor watcher. The results of this study raise the question of whether improved accuracy of detection of dysrhythmias results in better outcomes for patients. PMID- 9215430 TI - Effect of dedicated monitor watchers on patients' outcomes. AB - BACKGROUND: In 55% of progressive care units, someone is assigned to watch the cardiac monitors at all times, but the effect of this practice on patients' outcomes has not been examined. OBJECTIVE: To evaluate the effect of continual observation of telemetry units by a monitor watcher on mortality, frequency of transfer to a critical care unit, and the occurrence of five life-threatening dysrhythmias. METHODS: Data for this quasi-experimental study were collected on 1185 patients for a 9-month period in 1993 when the cardiac progressive care unit had a monitor watcher and on 1198 patients for a 9-month period in 1994 when the unit had no monitor watcher. RESULTS: We found no significant differences in mortality, frequency of transfer to a critical care unit, or the occurrence of three of the five dysrhythmias examined. The presence of a monitor watcher was associated with significantly fewer episodes of sustained ventricular tachycardia but more bradyarrhythmias. For both sustained ventricular tachycardia and bradyarrhythmias, the monitor watcher variable remained in the final multivariate logistic regression models. CONCLUSIONS: The presence of a monitor watcher was not associated with lower rates of most adverse outcomes evaluated; however, fewer episodes of sustained ventricular tachycardia occurred when a monitor watcher was present. Sustained ventricular tachycardia is life-threatening, disturbing to the patient, and may result in a longer hospital stay while medical therapy is being adjusted. The results of this study support the use of a monitor watcher to prevent sustained ventricular tachycardia. PMID- 9215431 TI - Comparison of methods of measuring pulmonary artery pressure. AB - BACKGROUND: Pulmonary artery waveforms fluctuate because of changes in intrathoracic pressure caused by respirations. Monitoring system algorithms determine digital displays of pressure measurements on the basis of recognition, analysis, and comparison of consecutive waveforms. OBJECTIVE: To compare three methods of measuring pulmonary artery pressure during mechanical ventilation and spontaneous breathing in cardiac surgery patients with stable hemodynamics. METHODS: Pulmonary artery pressure was measured during mechanical ventilation after cardiac surgery in 53 patients; 37 of the patients were studied again after extubation. Three monitoring methods were compared: graphic strip recording, the "stop cursor" (monitor screen freezing) method, and digital-display recording. Difference scores were calculated between the methods and analyzed for frequency and direction. RESULTS: All comparisons showed differences of at least +/-3 mm Hg in measurements obtained with the three methods. During mechanical ventilation, the digital and graphic measurements of systolic pressure varied most often; 57% (30/53) of the comparisons had difference scores of at least +/-3 mm Hg. The cursor and graphic measurements of diastolic pressures varied least often; 6% (3/53) of the comparisons had difference scores of at least +/-3 mm Hg. As expected, the digital method most often gave higher results than the graphic method. During spontaneous breathing, measurements of systolic pressure varied more often (38% to 53%) than did measurements of diastolic pressure (12% to 37%). Unexpectedly, for systolic pressures, the difference between digital and graphic measurements was 3 mm Hg or more 30% (11/37) of the time, and the difference between cursor and graphic measurements was 3 mm Hg or more 53% (17/32) of the time. CONCLUSIONS: Because of physiological and technical influences, measurements of systolic and diastolic pressures in the pulmonary artery made with the digital and cursor methods were not as reliable as measurements made with the graphic method. The findings support continued use of the graphic method for accurate measurements of pulmonary artery pressure. PMID- 9215432 TI - Methodological challenges to the study of occupational injury--an international epidemiology workshop. AB - Occupational and work-related injuries comprise the majority of reported workplace morbidity in the employed population in the United States. Despite intervention attempts, the overall trend for these injuries has been relatively stable over the past several decades. Three significant problems are raised as potentially contributing to this stability. Minimal progress in reduction may be due to: lack of etiologic understanding, lack of appropriate intervention selection, or lack of appropriate intervention implementation. Focusing on the first of these problems, a workshop symposium of injury epidemiologists and related scientists was held that developed a series of collaborative manuscripts on issues in injury epidemiology and recommendations for future research. The contributions of each are briefly summarized. PMID- 9215433 TI - Conceptual and definitional issues in occupational injury epidemiology. AB - This paper presents several models that further define the concept of occupational injury. While traditional models have proved successful in isolating specific research questions and health phenomena, the conceptual model presented permits a broader view of all injury morbidity. This model is based on both the level and frequency of energy transfers. A process model of occupational injury is also presented to describe the basic pathophysiological relationships associated with tissue effects/damage and recovery/repair. Numerous tradeoffs exist in variable selection, and a third model explores some of these tradeoffs. Differences in terminology and fundamental principles can limit the progress of occupational injury research. Accordingly, an argument is made for consolidation and consensus of terms. Finally, considerations for research are suggested, with an emphasis on the severity of the injury, the risk ratio, and the population at risk. PMID- 9215434 TI - Three perspectives on work-related injury surveillance systems. AB - This paper reviews surveillance approaches for occupational injuries and evaluates three emerging methodologies for the enhancement of work-related injury surveillance: (1) narrative data analysis, (2) data set linkage, and (3) comprehensive company-wide surveillance systems. All three methods are the result of new applications of computer hardware and software that have apparent strengths and limitations. A major strength is the improved description of work exposures and related injuries leading to better understanding of injury etiology. This understanding, however, is limited by the data quality and completeness entered on records at the time of the injury. We recommend (1) more widespread inclusion of narrative text in databases, analyses of which can be a valuable supplement to injury coded data; (2) the increased use of data set linkage studies to combine injury and work-history data; and (3) the development of comprehensive company-wide surveillance systems to expedite the use of epidemiologic data for occupational injury prevention activities. Further development of these methods and others is encouraged, especially in light of technological advancements in data capture, analysis and presentation. Only through such efforts can we best apply epidemiologic principles to preventing injuries in the workplace. PMID- 9215435 TI - Alternative approaches to analytical designs in occupational injury epidemiology. AB - In this paper, we discuss the theoretical framework upon which observational studies of occupational injuries are based. Following a general description of how causal effects are estimated, the challenges faced by researchers working in this area are outlined, with an emphasis on case-control studies. These challenges include defining the at-risk period for workers whose tasks change over time and whose hazard period may be very brief, evaluating the underreporting of both exposures and injuries, and considering the effects of multiple injuries per individual on study design and data analysis. We review both the theoretical and practical considerations in the design and conduct of traditional case-control studies, based on the collection of individual level data, as well as other approaches, such as using information culled from administrative and descriptive databases, and case-control studies in which the plant or work site is the unit of analysis. The case-crossover design is also reviewed and its utility for reducing confounding due to differences between individuals by self-matching is highlighted. While this design has not yet been applied to the work setting, its potential for increasing our understanding of the causes of acute-onset occupational injuries seems promising. Finally, a variety of hybrid designs are discussed, including combinations of case-control, case-crossover, and cohort designs. PMID- 9215436 TI - Challenges in assessing risk factors in epidemiologic studies on back disorders. AB - In epidemiologic studies on musculoskeletal disorders, some risk factors, especially physical load, cannot be determined independently from the worker. Posture, movement and external load are the result both of physical work requirements forced on the worker and of the worker's capacity to adopt particular techniques. Risk factors are also adjusted in relation to the worker's health. This paper presents a dynamic model that links exposure to risk factors for back pain and disability. Its aim is to help identify core elements in exposure assessment strategies for epidemiologic studies on back disorders. In this dynamic model, risk factors are determined relative to health status in order to distinguish between etiological and prognostic factors. Measurement techniques for various risk factors are classified into self-reports, observations, and direct instrumentation. Features of commonly used techniques are discussed with respect to feasibility, accuracy, and precision. In addition, consideration is given to the optimum allocation of measurements taking into account the effects of random and systematic variation in exposure due to tasks, workplaces, and workers. PMID- 9215437 TI - Design factors in epidemiologic cohort studies of work-related low back injury or pain. AB - The connection between work-related exposures and the onset of back injury or pain is complex and not clearly understood. This paper raises design issues related to the planning and conduct of cohort studies of industrial low back pain (or injury)(LBP), with care given to definition and measurement of exposure and outcome events. These issues include sample size, outcome definition, study biases, and practical considerations when seeking and maintaining company collaboration with a research effort. Without resolving these issues, the authors conclude: (1) cohort studies of worksite-based LBP are needed to elucidate the causal associations between work tasks and LBP onset, (2) both acute and cumulative exposures should be assessed as risk factors for low back injury or pain, and (3) attention should be paid to the planning of such studies and minimization of potential biases that can limit the validity of the results. These design issues will benefit researchers and companies engaged in the planning and conduct of cohort studies of industrial LBP. PMID- 9215439 TI - Advancing epidemiologic studies of occupational injury--approaches and future directions. AB - Despite increased attention to the epidemiology of occupational injuries and work related musculoskeletal disorders, little is known about the relative contribution of risk factors for specific injuries and diseases and effective ways to prevent them. Building on the knowledge and ideas presented in the previous papers in this special issue, this paper describes the salient issues regarding the needs and the opportunities in design, conduct, and analysis of epidemiologic research on occupational injury. There is a clear need for standardization of definitions on exposure variables and health outcomes, since basic terminology is not used consistently across studies. Improved surveillance systems with linkage to other databases offer excellent opportunities to evaluate trends in risk factors and occupational injuries. New opportunities for etiologic studies include the case-crossover design, which focuses on relations between intermittent exposures and injuries, as well as several hybrid designs that combine advantages and strength of the traditional case-control and cohort study designs. Various suggestions are presented to capture the dynamics of exposure, including both intermittent and continuous factors in the assessment strategy. The importance of cohort studies is emphasized to clarify the temporal relations between causes and effects in work-related injury research. Finally, intervention studies are much needed and a hierarchical approach is suggested, starting with observational surveys, continuing with quasi-experimental studies and concluding with randomized controlled experiments of those interventions that hold the largest promise. PMID- 9215438 TI - Design and conduct of occupational injury intervention studies: a review of evaluation strategies. AB - Occupational injuries continue to exact a great toll on American workers and their employers--the physical and financial costs are enormous. However, in the current political climate, few employers or regulatory agencies will implement injury prevention interventions without specific evidence of their effectiveness. This paper reviews the literature on the design, conduct, and evaluation of occupational injury interventions. Our review suggests that randomized controlled trials are rare and also notes that the quasi-experimental studies in the literature often use the weakest designs. We recommend a hierarchical approach to evaluating occupational injury interventions--beginning with qualitative studies, following up with simple quasi-experimental designs using historical controls, continuing with more elaborate quasi-experimental designs comparing different firms' experience, and, when necessary, implementing randomized controlled trials. PMID- 9215440 TI - Simultaneous diuresis cysto-urethrometry and multi-channel urethral pressure profilometry in female dogs with refractory urinary incontinence. AB - OBJECTIVE: To assess urethral closure and bladder storage function in female dogs with refractory urinary incontinence by use of simultaneous diuresis cysto urethrometry (UCM) and multi-channel urethral pressure profilometry (UPP). ANIMALS: 53 female dogs of various breeds with a history of 'non-neurogenic,' acquired urinary incontinence unresponsive to standard conservative treatment. PROCEDURE: UPP and UCM were performed on dogs sedated with xylazine, using a flexible polyvinylchloride multi-channel catheter connected to a perfusion system. Urine production was determined by use of radionucleotide dilution analysis. Values of reproducible urodynamic variables obtained previously in clinically normal dogs were compared with findings in the incontinent dogs. Cutoff values with the highest predictive values of urodynamic variables were determined and selected for functional classification. RESULTS: Except for threshold volume, median values of all urodynamic variables were significantly different between continent and incontinent dogs. Urethral pressure values, their relative variation, bladder threshold pressure, and prevalence of detrusor contractions were significantly lower than values in normal dogs, but compliance was higher. Forty-one dogs had urodynamic findings suggestive of poor urethral closure function assessed by UCM, but 10 of these had normal closure pressure values during UPP. Abnormal bladder storage function was interpreted from the findings of 19 dogs. Low capacity was found in 3, low threshold pressure in 15, and high compliance in 9 dogs. Seven of the dogs with abnormal bladder storage function had normal urethral closure function assessed by UPP and UCM. Five dogs had no abnormal findings, and 1 continent dog was abnormal by our classification. CONCLUSIONS: Abnormal bladder storage function was suggested in a considerable number of dogs with refractory urinary incontinence. PMID- 9215441 TI - Use of Doppler ultrasonography to evaluate renal arterial blood flow in horses. AB - OBJECTIVE: To obtain Doppler ultrasonographic images of renal arteries in horses and to establish reference range values for systolic and diastolic renal arterial blood flow and resistive indices. Also to determine whether Doppler ultrasonography could be used in horses to detect changes in renal blood flow after IV administration of furosemide. ANIMALS: 11 clinically normal adult horses. PROCEDURES: Pulsed-wave Doppler examinations were performed on arcuate arteries of 5 sedated horses. Continuous-wave Doppler examinations were performed on pyelorenal arteries in 7 nonsedated horses and were repeated in 6 horses after furosemide administration. Peak-systolic velocity (SV) and end-diastolic velocity (EDV) were measured and the resistive indices (RI) were calculated. RESULTS: Using pulse-wave Doppler ultrasonography in sedated horses, arcuate arteries were determined to have a SV of 0.406 +/- 0.116 m/s (mean +/- SD), EDV of 0.184 +/- 0.057 m/s, and RI of 0.549 +/- 0.044. Using continuous-wave Doppler ultrasonography in nonsedated horses, pyelorenal arteries were determined to have SV of 1.047 +/- 0.009 m/s, EDV of 0.510 +/- 0.006 m/s, and RI of 0.512 +/- 0.004. Doppler waveforms from the arcuate and pyelorenal arteries had a low resistance flow pattern, characterized by a systolic peak followed by a continuous antegrade diastolic flow. After furosemide administration, the pyelorenal arterial velocities increased, but the RI remained unchanged. CONCLUSIONS: Doppler ultrasonography may be used to record renal blood flow in horses and to detect changes following furosemide administration. CLINICAL RELEVANCE: Doppler ultrasonographic images may assist in the diagnosis of renal diseases that affect either blood flow or Doppler waveform. PMID- 9215442 TI - Multiplex polymerase chain reaction assay for genotyping Clostridium perfringens. AB - OBJECTIVE: To develop a multiplex polymerase chain reaction (PCR) assay to detect the genes for the major toxins of Clostridium perfringens (cpa [alpha toxin], cpb [beta toxin], etx [epsilon toxin], iA [iota toxin], and cpe [enterotoxin]). SAMPLE POPULATION: Cultures of C perfringens obtained from collections and diagnosticians throughout North America. PROCEDURE: PCR primers were derived from published sequences of the genes for the major toxins (the "typing" toxins and enterotoxin). The concentration of each primer was titrated in a PCR assay to allow concurrent amplification of multiple target sequences, and other parameters of the assay were optimized (including concentrations of other reagents and times and temperatures for denaturation of template, annealing of primers, and primer extension). Specificity of the assay was measured by comparing genotype with phenotype (where it was known). RESULTS: The genotype, determined by multiplex PCR assay, agreed with phenotype in 99% (86/87) of strains where phenotype had been determined. Applied to 361 isolates from domestic animals and human beings, 95% (n = 344) were type A, and 12.8% (n = 44) of these contained cpe. The remaining 5% (n = 17) of the isolates were type B (n = 1), type C (n = 11), type D (n = 2), or type E (n = 4). CONCLUSION AND CLINICAL RELEVANCE: Previous studies have documented usefulness of PCR in genotyping C perfringens. The multiplex assay is as effective, but simpler, and may be a useful alternative to standard in vivo typing methods. Results of genotyping of field isolates suggested the need for further epidemiologic study of clostridial enteritis, particularly as this pertains to predominant etiologic toxin types, and documented the presence of the reportedly rare genotypes B and E. PMID- 9215443 TI - Profile of electrodiagnostic abnormalities in cats with GM1 gangliosidosis. AB - OBJECTIVE: To determine which electrodiagnostic tests yield abnormal findings in cats with GM1 gangliosidosis, and to determine the approximate age of onset of electrodiagnostic abnormalities. ANIMALS: Cats (28 to 335 days old) affected with GM1 gangliosidosis (n = 11) and unaffected controls (n = 14). PROCEDURE: Cats were grouped by age: group 1, < or = 90 days, group 2, 91 to 200 days; and group 3, > 200 days. Electrodiagnostic tests were conducted, including needle electromyography, motor and sensory nerve conduction velocity, spinal evoked potentials, and brainstem auditory evoked potentials. Results for control and affected cats were compared, using the general linear model for ANOVA and Scheffe's test for multiple comparisons. RESULTS: Needle electromyography did not reveal abnormal spontaneous activity in skeletal muscles of any cat; furthermore, statistical analysis did not indicate significant difference between affected and control groups for nerve conduction velocity, confirming that degeneration of peripheral nerve fibers is not a feature of this disease. However, spinal evoked potentials were abnormal in group-3 cats; conduction velocity within sensory pathways in the cranial part of the spinal cord was significantly slower in GM1 affected cats (P = 0.0002). Brainstem auditory evoked responses also were abnormal: wave V (generated in the region of the pons) had prolonged latency in cats of groups 2 and 3 (P = 0.0003 and 0.0001, respectively, at 90 decibels sound pressure level). In the oldest cats, latencies for earlier waves within the auditory pathway also were prolonged; wave I (generated by the cochlear nerve) was prolonged in group-3 cats (P = 0.0423). CONCLUSIONS: Motor and sensory nerve conduction velocities remained within normal limits in GM1-affected cats. However, spinal evoked potentials indicated slowing in conduction velocity along the cranial part of the spinal cord in group 3 cats. Brainstem auditory evoked responses indicated prolonged latencies in cats of groups 2 and 3. PMID- 9215444 TI - Characterization of anatomic communications of the fetlock in cattle, using intra articular latex injection and positive-contrast arthrography. AB - OBJECTIVE: To evaluate the frequency and sites of communication between the lateral and medial synovial sacs of the metatarsophalangeal or metacarpophalangeal joints in cattle. ANIMALS: 188 limbs were obtained from 55 fresh bovine cadavers submitted for necropsy because of problems unrelated to the fetlocks. PROCEDURE: In each ox, lateral or medial synovial sacs of each fetlock were randomly assigned. Joints were injected with a mixture of latex and barium sulfate. Communication between 2 joints was determined by presence of latex and contrast material in a joint adjacent to the injected joint by examining frozen sections and use of positive-contrast arthrography. RESULTS: Communication between the 2 synovial sacs existed in 186 of 188 (98.9%) specimens. The communication site between lateral and medial synovial sacs was located at the level of the proximal sesamoid bones, between the distal aspect of the interdigital band of the axial branch of the interosseus muscle and the metacarpal or metatarsal bone. CONCLUSIONS AND CLINICAL RELEVANCE: Although communication between the lateral and medial synovial sacs did not exist in 2 specimens, the fetlock in cattle can be treated as 1 compartment. PMID- 9215445 TI - Modeling study of compensatory head movements in lame horses. AB - OBJECTIVE: To study the role of head movements in lame horses. SAMPLE POPULATION: 11 Dutch Warmblood horses. PROCEDURE: A 2-segment 2-dimensional inverse dynamic model of trotting horses was developed: trunk and head/neck segment joined in a neck joint. Model input consisted of averaged segmental inertial properties and averaged kinematic data, taken from 11 horses, trotting on a treadmill (3.5 m/s) in 3 conditions of induced lameness: sound, mildly lame, and moderately lame. Dynamic and static effects were analyzed. RESULTS: Dynamic effects were found to be considerably larger than static effects. In the moderately lame condition, the maximal neck joint vertical force during the lame stance phase had a 27% decrease, compared with the sound situation. Neck joint sagittal torque and maximal vertical force on the trunk decreased by 31 and 13%, respectively. Load distribution between forelimb and hind limb indicated a relative load shift from the lame forelimb to the diagonal hind limb during the lame stance phase. The sound contralateral forelimb carried a higher load while the ipsilateral hind limb was unloaded. CONCLUSION: It could be concluded that asymmetric head movements have a major role in lameness compensation, which can be explained by inertial interaction between trunk and head/neck segment. Static effects, such as caudad shifting of the body center of mass, are of minor importance. CLINICAL RELEVANCE: This report clarifies the mechanism of lameness compensation and the method of lameness diagnosis. PMID- 9215446 TI - Repeated physical and cytologic characterizations of subcutaneous postvaccinal reactions in cats. AB - OBJECTIVES: To examine local reactions and short-term cytologic responses of cats to administration of rabies virus (RV); FeLV; and combined feline rhinotracheitis, calicivirus, and panleukopenia virus (FRCPV) vaccines. ANIMALS: 9 healthy 6- to 7-month-old specific-pathogen-free cats. PROCEDURE: One milliliter doses of the aforementioned vaccines were administered SC (at different sites) to healthy, specific-pathogen-free cats. Each cat also received 1 ml of sterile saline solution SC as a control. Injection sites were visually examined and palpated daily for 4 weeks. Palpable lesions were measured by use of calipers. Temperature of the vaccination sites was measured weekly by use of a thermocouple. Aspirates were taken from vaccination sites weekly, and smears were submitted for cytologic analysis. RESULTS: There were no significant differences in lesion surface temperature among injection sites at any time. Injections of saline solution and FeLV vaccine resulted in no palpable lesions. The FRCPV vaccine elicited a minor reaction in 1 of the 9 cats. The RV vaccine caused palpable lesions in all cats. Smears of the aspirates from the sites of saline injection were poorly cellular. Cellularity of aspirates from the sites of FRCPV and FeLV vaccinations was moderate at week 1, and decreased with time. Inflammatory infiltrates were composed principally of lymphocytes, with fewer neutrophils and macrophages. In contrast, cellularity of aspirates from RV vaccination sites increased for 21 days and was characterized by increasing numbers of lymphocytes and macrophages. CONCLUSIONS: RV vaccine used in this study induced palpable lesions in many cats. In contrast, FRCPV and FeLV vaccines elicited less severe lesions. CLINICAL RELEVANCE: Subcutaneous administration of killed virus vaccines in cats may result in palpable lesions that are detected by clients or clinicians. Aspiration cytologic examination may reveal a different characteristic pattern of cells that is dependent on the individual vaccine and time elapsed from vaccination. PMID- 9215448 TI - Hyaluronate concentration in tracheal lavage fluid from clinically normal horses and horses with chronic obstructive pulmonary disease. AB - OBJECTIVE: To establish concentration of hyaluronate (HA) in tracheal lavage fluid from healthy horses and horses with chronic obstructive pulmonary disease (COPD). ANIMALS AND SAMPLES: Tracheal lavage fluid samples (n = 42) from 18 horses, 11 with COPD, and 7 control horses. PROCEDURE: Clinical examination of the respiratory tract, tracheal lavage, and blood sample collection were performed on horses without clinical signs of respiratory tract disease and horses with clinical signs of COPD. In some horses, 1 to 5 repeated examinations were performed at 1-week intervals. Tracheal lavage fluid samples were analyzed for cell numbers, and urea concentration (made in parallel with serum samples to evaluate sample dilution effect); HA was determined by radiometric assay. RESULTS: Mean (+/-SEM) HA concentration in tracheal lavage fluid samples was significantly (P = 0.005) higher in horses with COPD (1,880 [+/-309] micrograms/L), compared with that in control horses (256 [+/-72] micrograms/L). The increase in HA concentration in tracheal lavage fluid of COPD-affected horses was verified by repeated sample collection and analysis. CONCLUSIONS: In horses with chronic respiratory tract inflammation such as COPD, tracheal lavage fluid HA concentration is about 7 times higher than reference values. High HA concentration in the tracheal or bronchoalveolar lavage fluid may reflect pathophysiologic changes in connective tissue around bronchi and bronchioli, leading to continuous increased production of HA in horses with advanced forms of COPD. CLINICAL RELEVANCE: Determination of tracheal lavage fluid HA concentration may be used as a marker of chronic inflammatory changes in the COPD-affected lung. PMID- 9215447 TI - Immunohistochemical localization of alpha 1-acid glycoprotein in liver tissues of bovine fetuses, newborn calves, and sick or healthy adult cattle. AB - OBJECTIVE: To detect localization of alpha 1-acid glycoprotein (alpha 1-AG) antigens in the liver tissue of cattle by use of immunoperoxidase technique. SAMPLE POPULATION: Liver specimens from 6 bovine fetuses, 2 healthy bovine neonates, 2 healthy adult cattle, 3 cattle with experimentally induced hepatic abscesses, and 2 cattle with enzootic bovine leukosis (EBL). PROCEDURE: 3 cattle (with hepatic abscesses) were inoculated with a suspension of Fusobacterium necrophorum in the ruminal vein. Serum alpha 1-AG concentration was determined by use of the single radial immunodiffusion method. Livers from fetuses, newborn calves, and adult or sick cattle were fixed in buffered 10% formalin, dehydrated in alcohol, embedded in paraffin, sectioned, and stained by use of the avidinbiotin complex/immunoperoxidase technique. RESULTS: Sites of localization of the alpha 1-AG antigen positive reaction (AGPR) in the liver obtained from bovine fetuses, neonates, or sick cattle were different. In fetal and newborn calves, the AGPR was detected in the cytoplasm of hepatocytes. Intensity of the reaction varied in direct proportion to alpha 1-AG serum concentration. In adult cattle, the AGPR was particularly intense in hepatocytes adjacent to abscesses or EBL-induced tumors. CONCLUSIONS: The pattern of distribution of cells with AGPR in the liver varied, depending on severity of inflammation. In the cattle with EBL, whether the AGPR was attributable to inflammation could not be clarified, although suppression of immunologic response to tumors may have been a cause of the observed reaction. This association suggests that the glycoprotein may be synthesized, mainly in hepatocytes. PMID- 9215449 TI - Investigation of a listeriosis epizootic in sheep in New York state. AB - OBJECTIVE: To investigate potential sources of an epizootic of listerial encephalitis, using molecular diagnostic and typing methods. SAMPLE POPULATION: A flock of about 655 sheep. PROCEDURE: An epizootiologic investigation was performed. Clinical, feed, and environmental samples were tested for Listeria monocytogenes, using polymerase chain reaction and culture methods; recovered isolates were "fingerprinted," using an automated ribotyping system. RESULTS: Listeria monocytogenes was recovered from brain specimens of 7 sheep with clinical signs of listerial encephalitis. All clinical isolates had fingerprints identical to those of isolates from farm equipment used to transport silage. Corn silage, which was not fed to the sheep, also contained L monocytogenes of the same pattern type as defined by ribotyping. Listeria monocytogenes was not isolated from the stored haylage designated for feeding the sheep (the cut-off point for isolation being < 10(2) colony-forming units/g). CONCLUSIONS: Corn silage was implicated as the source of a listeriosis epizootic. It appears to have cross-contaminated the haylage destined for the sheep during handling with a front-end loader. Suspension of silage feeding coincided with cessation of listeriosis cases. CLINICAL RELEVANCE: Use of advanced molecular techniques can help to identify the sources and restrict the scope of an epizootic. In epizootics, a single L monocytogenes strain can lead to infection of multiple animals, with rapid progression of the disease. PMID- 9215450 TI - Influences of breed, sex, and susceptibility to malignant hyperthermia on lipid composition of skeletal muscle and adipose tissue in swine. AB - OBJECTIVE: To determine influences of breed, sex, and susceptibility to malignant hyperthermia on composition of skeletal muscle and adipose tissue in swine. ANIMALS: 35 male and female swine of German Landrace and Pietrain breeds and of 2 genotypes, normal (MHN) and susceptible to malignant hyperthermia (MHS). PROCEDURE: Pigs were fed a standard diet ad libitum. After attaining body weight of approximately 100 kg, pigs were slaughtered and skeletal (longissimus thoracis and supraspinatus) muscle and adipose tissue (3 sites) specimens were removed. For each specimen, lipids were extracted by chloroform/methanol and fatty acid (FA) pattern, and cholesterol concentration was determined by gas chromatography. RESULTS: The overall lipid contents differed significantly between breeds and genotypes; the MHS Pietrain pigs had the lowest lipid quantities. The relative amounts of saturated FA in all tissues were highest in Landrace pigs (P < 0.05), whereas the relative contents of monoenic FA were lower. In addition, for both breeds, the MHN pigs had significantly higher saturated and lower polyunsaturated FA values in all tissues, compared with MHS pigs. More specifically, MHS females of both breeds had the highest relative amounts of polyunsaturated FA. In general, relative cholesterol contents were found to vary little between identified groups. CONCLUSIONS: These data may indicate that, not only does mutation of the calcium release channel of the sarcoplasmatic reticulum, which occurs in MHS swine, influence secondary changes in lipid composition, but so do hormone concentrations and other genotypic factors. Observed differences in lipid content and FA composition could consequently influence specific membrane properties, such as fluidity and cell signaling. PMID- 9215451 TI - Humoral response of dairy cattle to spirochetes isolated from papillomatous digital dermatitis lesions. AB - OBJECTIVE: To determine whether a humoral response against spirochetes isolated from papillomatous digital dermatitis (PDD) lesions is elicited in dairy cattle affected with PDD. SAMPLE POPULATION: 41 cattle with PDD from 8 dairies (study population) and 30 cattle from 2 dairies free of PDD (control population). Additionally evaluated were 32 cattle from a dairy with a past history of PDD but no current disease, and 52 cattle from a dairy with high prevalence of PDD, 25 with and 27 without detectable lesions. PROCEDURE: ELISA were used to evaluate the humoral response of all cattle to representative isolates from 2 groups of spirochetes of unknown species isolated from PDD lesions. Specificity of the response was evaluated, using immune sera prepared against each of the spirochetes, and by adsorption studies of immune and field sera. The potential for confounding by an antibody response to other spirochetes associated with diseases of cattle was assessed. RESULTS: The antibody response (specific) to both PDD spirochete groups of cows with PDD was significantly increased, compared with that of cows from PDD-free dairies. There was no association between antibody response to PDD-associated spirochetes and antibody response to other spirochetal diseases of cattle. None of the cattle from the dairy with previous history of PDD but without current disease were classified as test positive by either PDD ELISA. There was a significant (P < 0.01) difference in classification results for both PDD ELISA for cattle with PDD from the dairy with a high herd prevalence of PDD, compared with cattle without detectable disease from the same dairy. CONCLUSIONS AND CLINICAL RELEVANCE: The humoral response in cattle with PDD lesions was significantly different from that in cattle without detectable lesions, thus providing additional information regarding the potential role of spirochetes isolated from PDD lesions in the etiopathogenesis of PDD. PMID- 9215452 TI - Growth of Pasteurella haemolytica and production of its leukotoxin in semi defined media. AB - OBJECTIVE: To develop semi-defined media that support growth of the bovine pathogen, Pasteurella haemolytica, and use them to examine production of leukotoxin and an arginine-binding protein by this organism. SAMPLE POPULATION: 10 P haemolytica A1 strains and 1 P multocida strain. PROCEDURE: Bacterial strains were cultivated at 37 C in media containing various amino acids, carbon sources, vitamins, and cofactors, and absorbance (OD600) was measured. Leukotoxin and arginine-binding protein production were assessed by immunoblot analysis. RESULTS: Optimal growth required supplementation with 0.1% fetal bovine serum, gelatin, or purified bovine serum albumin. Calcium pantothenate and thiamine were essential for growth, and a variety of carbon sources could be utilized. In the complete medium, 15 amino acids were included; however, in the minimal medium, no amino acids were required. All strains (except P multocida) grew in the complete medium and 7 grew well in the minimal medium. Leukotoxin was not produced when amino acids were limiting, but could be enhanced by addition of 0.2% NH4SO4. Production of the arginine-binding protein was not affected by nitrogen availability or by presence of L-arginine. CONCLUSIONS: Two media that support good growth of P haemolytica strains were developed. The minimal medium is simple to prepare and manipulate and its use revealed a potential role of nitrogen availability in the regulation of leukotoxin expression. CLINICAL RELEVANCE: Creation of these media will permit continued studies of the response of P haemolytica to environmental conditions that may mimic those encountered in the bovine respiratory tract during shipping. PMID- 9215453 TI - Intrastrain variation of lipopolysaccharide of Pasteurella multocida in turkeys. AB - OBJECTIVE: To document intrastrain variation of lipopolysaccharide (LPS) in Pasteurella multocida and correlate these changes with changes in determinants associated with virulence. ANIMALS: 25 broad-breasted white turkeys. PROCEDURE: Phenotypic bacterial variants were identified by lectin affinity and were assayed for adherence to epithelial cells and complement resistance in vitro. The LPS purified from these variants was subjected to polyacrylamide gel electrophoresis and lectin affinity analysis. Turkeys were challenge exposed, then observed for 1 week. At first sign of disease, or at the end of the study, turkeys were euthanatized, necropsied, and inspected for gross lesions. RESULTS: The LPS variant designated as Ricinus communis agglutinin-positive had greater adherence to epithelial cells, complement resistance, and virulence in turkeys than did the variant designated as R communis agglutinin-negative. CONCLUSIONS: Intrastrain variation of LPS exists in P multocida, and changes in LPS are correlated with changes in virulence. PMID- 9215454 TI - Physical, hematologic, biochemical, and immunologic effects of supranutritional supplementation with dietary selenium in Holstein cows. AB - OBJECTIVE: To measure responses of cows to supplemental Se intake in excess of nutritional requirements, but lower than recognized toxic dosages. ANIMALS: 24 healthy adult Holstein cows. PROCEDURE: Cows were allotted to 4 groups and fed sodium selenite to provide 0, 3, 20, or 50 mg of Se/cow/d for 90 days. Subsequently, the dosage for the group receiving 50 mg/cow/d was increased to 100 mg/cow/d for 28 d. Blood, liver specimens, feces, and urine were obtained at points during the trial. RESULTS: Serum and blood Se concentrations in groups receiving 20 or 50 mg/cow/d increased over time, compared with controls (P < 0.01). Increasing supplemental Se intake to 100 mg/cow/d further increased serum and blood Se concentrations (P < 0.05). Urine, fecal, and liver Se concentrations increased more markedly in response to treatment than did those of serum or blood. No effect of Se treatment was seen on blood cell counts or serum activities of hepatocellular enzymes. Likewise, neither titer response to rabies vaccination nor lymphocyte blastogenic response to nonspecific mitogens was affected by Se treatment. Objective or subjective physical signs of Se toxicosis were not observed at any Se dosage. CONCLUSIONS: Inorganic Se intakes of up to 50 mg/d for 90 days or 100 mg/d for 28 days by adult Holstein cows do not affect the variables measured. CLINICAL RELEVANCE: Intakes of Se as sodium selenite in amounts 10 to 30 times the nutritional requirements are unlikely to cause health problems in adult cows. Urine and feces are good test samples for detection of Se supplementation greater than requirements. PMID- 9215456 TI - Acid-base, metabolic, and hemodynamic effects of sodium bicarbonate or tromethamine administration in anesthetized dogs with experimentally induced metabolic acidosis. AB - OBJECTIVE: To evaluate buffering capacity and side effects of equivalent doses of tromethamine (THAM) and sodium bicarbonate (BIC). ANIMALS: 18 purebred dogs. PROCEDURE: Acidosis was induced by having dogs breathe a hypoxic gas mixture (FIO2 = 0.10) until arterial base balance < or = -7.5 mEq/L was reached. Dogs then received a 30-minute infusion of 5% BIC (n = 6) or 0.3M THAM (n = 8), and FIO2 increased to 0.30. Drug doses were calculated to correct base balance to zero. RESULTS: During hypoxia, for BIC- and THAM-treated groups, median (interquartile range [Q1, Q3]) pHa and arterial base balance decreased to 7.16 (7.07, 7.38) and 7.19 (7.11, 7.31), -14 (-16, 9) and -12 (-16, -11) mEq/L, respectively, and mixed venous lactate concentration increased to 7 (2, 15) and 6 (3, 13) mmol/L, respectively. Immediately after each infusion, acid-base and cardiopulmonary variables returned toward baseline. For respective BIC- and THAM treated groups, pHa increased to 7.37 (7.26, 7.44) and 7.40 (7.33, 7.49) and base balance increased to 0 (-4, 7) and 0 (-4, 2) mEq/L. Lactate concentration decreased only slightly to 5 (2, 6) and 5 (2, 9) mmol/L, but continued to decrease throughout the study. The only significant (P < or = 0.05) difference between groups was hypernatremia after BIC administration that persisted for 60 minutes. The PaCO2 in BIC-treated dogs increased immediately after infusion, compared with values during hypoxia. Standardized ionized calcium values initially decreased in both groups, but returned to baseline by 60 minutes. CONCLUSION: The buffering capacity of THAM is equal to that of BIC, although THAM does not cause the transient hypernatremia or hypercapnia observed after BIC administration. Hypocalcemia may be transient after administration of either solution. Thus, THAM is an acceptable alternative to BIC for treatment of metabolic acidosis in selected anesthetized dogs. PMID- 9215455 TI - Endocrine changes in cerebrospinal fluid, pituitary effluent, and peripheral plasma of anesthetized ponies. AB - OBJECTIVE: To investigate the effects of inhalation and total IV anesthesia on pituitary-adrenal activity in ponies. ANIMALS: 9 healthy ponies: 5 geldings and 4 mares. PROCEDURE: Catheters were placed in the cavernous sinus below the pituitary gland and in the subarachnoid space via the lumbosacral space. After 72 hours, administration of acepromazine was followed by induction of anesthesia with thiopentone and maintenance with halothane (halothane protocol), or for the IV protocol, anesthesia induction with detomidine and ketamine was followed by maintenance with IV infusion of a detomidine-ketamine-guaifenesin combination. Arterial blood pressure and gas tensions were measured throughout anesthesia. Peptide and catecholamine concentrations were measured in pituitary effluent, peripheral plasma, and CSF. Peripheral plasma cortisol, glucose, and lactate concentrations also were measured. RESULTS: Intravenous anesthesia caused less cardiorespiratory depression than did halothane. ACTH, metenkephalin, arginine vasopressin, and norepinephrine pituitary effluent and peripheral plasma concentrations were higher during halothane anesthesia, with little change during intravenous anesthesia. Pituitary effluent plasma beta-endorphin and peripheral plasma cortisol concentrations increased during halothane anesthesia only. Dynorphin concentrations did not change in either group. Hyperglycemia developed during intravenous anesthesia only. Minimal changes occurred in CSF hormonal concentrations during anesthesia. CONCLUSION: The pituitary gland has a major role in maintaining circulating peptides during anesthesia. Compared with halothane, IV anesthesia appeared to suppress pituitary secretion. PMID- 9215457 TI - Effects of tromethamine buffer on coagulation variables and ionized calcium concentration in dogs. AB - OBJECTIVE: To evaluate coagulation variables in 2 groups of dogs after tromethamine administration. ANIMALS: 13 Beagles. PROCEDURES: Both groups of dogs received a 30-minute IV infusion of 10 ml of 0.3M tromethamine/kg of body weight. In unsedated dogs (group 1, n = 8), prothrombin time, activated partial thromboplastin time, normalized ionized calcium concentration, platelet numbers, and platelet function were measured prior to treatment, at the end of the infusion, and 1 hour after the infusion. In xylazine-sedated dogs (group 2, n = 5), buccal mucosal bleeding time and plasma percentage of von Willebrand factor antigen were measured before and 1 hour after infusion, and fibrin degradation products concentration was measured 1 hour after infusion. Platelet function was assessed by determining platelet aggregation and by measuring ATP release from the aggregating platelets over 6 minutes, using a whole blood aggregometer, with 20, 10, and 5 microM ADP and 5 and 10 micrograms of collagen/ml as platelet activation agonists. RESULTS: There was no significant change in any of the variables measured in either group of dogs, compared with baseline values. CONCLUSIONS AND CLINICAL RELEVANCE: When administered to healthy dogs, tromethamine does not change the coagulation indices measured. PMID- 9215458 TI - Distribution of technetium 99m-labeled red blood cells during isoflurane anesthesia in ferrets. AB - OBJECTIVE: To address the physiologic mechanism of isoflurane-associated reduction in hematologic variables in ferrets. ANIMALS: 6 young adult female ferrets. PROCEDURE: Distribution of 99mTc-labeled autologous erythrocytes was measured by serial in vivo imaging. Data were recorded in 4 ferrets, using a gamma camera, immediately prior to anesthesia, 15 minutes after 2% isoflurane anesthesia in O2 via endotracheal tube, 1 minute prior to and throughout a 10 minute phenylephrine infusion, 20 and 40 minutes after termination of the phenylephrine infusion, and 45 minutes after termination of anesthesia. Blood indices were also measured at times that paralleled those for imaging. One ferret served as a conscious control (no anesthetic administration), and another as an isoflurane control (no phenylephrine administration). RESULTS: In ferrets under anesthesia, splenic radioactivity increased from baseline of 10.2 +/- 2.0% to 38.4 +/- 3.2% (mean +/- SEM; P < 0.05) of the injected dose. Splenic radioactivity decreased to 13.4 +/- 3.8% of the injected dose during phenylephrine infusion and to near baseline for the recovery image. Splenic radioactivity in the conscious control remained constant throughout the study, whereas that of the anesthetized control was persistently increased throughout administration of isoflurane. Percentage reduction of the 15-minute sample values, compared with baseline values for all hematologic indices, was: RBC count, 33% (P < 0.05); hemoglobin concentration, 34% (P < 0.05); hematocrit, 35% (P < 0.05); and plasma protein concentration, 20% (P < 0.05). All RBC variables returned to within 7 to 14% of baseline by 45 minutes after termination of anesthesia. CONCLUSION: Isoflurane anesthesia causes splenic sequestration of RBC in ferrets that is partially reversed by phenylephrine infusion or termination of anesthesia. Thus, investigators and clinicians should be cautious when interpreting hematologic findings in isoflurane-anesthetized ferrets, and accordingly, fluid treatment and transfusion should be planned. PMID- 9215459 TI - Craniad migration of differing doses of new methylene blue injected into the epidural space after death of calves and juvenile pigs. AB - OBJECTIVE: To determine the relation between epidural injectate volume (ml/kg of body weight) and its craniad migration in calves and pigs. ANIMALS: 23 neonatal calves and 26 feeder pigs. PROCEDURE: Animals were randomly assigned to receive different volumes of new methylene blue (NMB, 1.2 mg/ml in 0.9% saline solution). Injections were made into the sacrococcygeal intervertebral space in calves and the lumbosacral intervertebral space in pigs, immediately after euthanasia. Sagittal sections of the spine were made at necropsy, and craniad migration of NMB was determined and rounded to the nearest intervertebral space. RESULTS: In calves treated with 0.05, 0.1, or 0.15 ml of NMB/kg, mean +/- SEM number of stained spinal segments was 5 +/- 0.3, 8 +/- 0.6, and 8 +/- 0.6, respectively. Craniad migration of NMB was significantly greater for 0.15 and 0.1 ml/kg volumes versus 0.05 ml/kg. In pigs treated with 0.05, 0.1, 0.2, of 0.3 ml of NMB/kg, mean number of stained spinal segments was 8 +/- 1.1, 8 +/- 0.9, 10 +/- 1.2, and 18 +/ 2.0. Craniad dye migration was significantly greater in the 0.3 ml/kg group versus the 3 lower volume groups. Linear regression performed on both sets of data after logarithmic transformation of spaces migrated to correct for non normality was significant (P < 0.05), and R2 values of 0.49 and 0.55 were obtained for calves and pigs, respectively. CONCLUSIONS: There is a significant correlation between volume (ml/kg) of NMB injected in the epidural space and its craniad migration in calves and pigs. CLINICAL RELEVANCE: Results provide a basis for determination of volume of injectate to be given to reach a minimal desired level and should be a useful baseline for future investigations of epidural drug administration. PMID- 9215460 TI - Evaluation of mid-term stability of night vision tests. AB - BACKGROUND: Dark adaptation rate, scotopic retinal sensitivity and contrast sensitivity under mesopic conditions, but not visual acuity, have been shown to be directly related to the ability to identify military targets at night. These parameters can be used to select personnel for specific military tasks demanding excellent night vision, as well as to assess pharmacological effects on night vision. PURPOSE: To evaluate the mid-term (2 to 6-week period) stability of night vision tests based on assessment of the above parameters. METHODS: Dark adaptation rate, scotopic retinal sensitivity and contrast sensitivity under mesopic conditions were studied in 16 young volunteers during a 6-week period. RESULTS: Tests of scotopic retinal sensitivity (after 30 min of dark adaptation) exhibited high reproducibility and a low fluctuation rate, with a high correlation between values at week 0 and at 2-week intervals during the following 6 weeks of the study (rs (week 0 to week 6) = 0.81, p = 0.0001). The reproducibility of mesopic contrast sensitivity tests (average of 1.5, 3, 6 and 12 cycles per degree, (cpd)) was fair (rs (week 0 to week 2) = 0.67, p = 0.0045), whereas that of dark adaptation rate tests was poor. CONCLUSIONS: In view of the reproducibility characteristics of these night vision tests, assessment of night vision ability in pilots and military personnel, as well as assessment of pharmacological effects on night vision, may be based on scotopic retinal sensitivity (after 30 min of dark adaptation) and contrast sensitivity under mesopic conditions (average of 1.5, 3, 6 and 12 cpd). PMID- 9215461 TI - Visual scanning and pilot expertise: the role of attentional flexibility and mental model development. AB - In order to examine differences in flying expertise, 12 novice and 12 expert pilots flew a 7-segment simulation pattern under specific attentional constraints while cockpit instrument visual scan was recorded. Flight segments involved various combinations of maneuvering of heading, altitude and airspeed. Expert pilots performed better than novices on vertical and longitudinal, but not lateral control. They accomplished their superior vertical tracking by allocating more control resources to the vertical control. Analyses of scanning strategies revealed that experts: a) had shorter dwells and more frequent visits to most instruments; b) adapted their visiting strategy more flexibly in response to changing task demands; c) demonstrated a better mental model of cross-coupling and predictive relations between and within axes; and d) showed more frequent checking of axes whose values remained constant. The data is discussed in terms of their implications in pilot cockpit scan training program development. PMID- 9215462 TI - Laboratory simulator and field pursuit tracking performance with females and males in the presence of laser glare. AB - BACKGROUND: The developments in laser technology have increased the precision of many tasks and has made the presence of lasers common-place. In the military the pervasive use of laser devices in uncontrolled environments enhances the potential for human exposure. The visual disruption experienced during these exposures could lead to serious injury or disruption of performance. Characterization of changes in visual-motor performance of military personnel exposed to safe levels of laser glare assists in minimizing mission performance decrements. METHODS: There were 18 female and male military personnel who performed a tracking task in the field and in the laboratory. Two systems were used to assess possible gender differences inherent to the operation of each unit. There were six, 3-s laser trials presented at an irradiance of 400 microW.cm-2 during 15 bright light and 15 simulated dawn/dusk trials with each system. The laser beam on the retina was collinear with the image on the sight. Maximum absolute error (MAE) and total time-off-target (TTOT) scores were determined. RESULTS: Analysis showed that after the flash females tended to lead and males lagged behind the target. No significant differences in MAE or TTOT scores attributable to gender were found. Dawn/dusk flash trials produced greater disruption of pursuit tracking than did bright light trials. Repeated flash exposures showed either an adaptive or a cumulative response. CONCLUSIONS: Significant visual disruption was found following exposure to "safe" levels of laser light and this effect was increased during simulated dawn/dusk conditions. The degree of performance decrement was not related to gender. PMID- 9215463 TI - Unusual vestibular and visual input in human dynamic balance as a motion sickness susceptibility test. AB - BACKGROUND: Motion sickness (MS) is commonly thought to arise from a sensory conflict. However, few quantitative methods based on this theory are available to detect MS susceptibility. HYPOTHESIS: It was asked whether the standardized unusual stimulation of a single sensory channel under quantified dynamic balance conditions in man could elicit a sensory conflict and therefore trigger motion sickness (MS) METHODS: Vestibular and visual channels were stimulated by galvanic current and rotating prismatic glasses, respectively. The moving platform used to create the requirements for dynamic balance conditions was chosen not only to worsen the malaise but also to obtain an objective measurement of the balance consequences of the stimulations. RESULTS: Both vestibular and visual stimulation, applied separately, elicited MS-like symptoms (in 56% and 73% of subjects, respectively) and stereotyped balance reactions. A relationship was found between subjective MS-like symptoms and objective measurements of dynamic balance performance. Subjects sensitive to unusual vestibular messages differed from the others by a greater increase in the parameters indicating a difficulty of balance whereas subjects sensitive to unusual visual messages were recognized by the strategy they used to balance themselves. CONCLUSIONS: These results demonstrated that a sensory conflict can trigger MS-like symptoms. We conclude that the measured parameters of a global somatomotor activity, such as the dynamic balance task proposed here, could be useful for objectively detecting subjects predisposed to MS, for training them and testing the efficiency of anti MS drugs. PMID- 9215464 TI - Echocardiographic findings in NATO pilots: do acceleration (+Gz) stresses damage the heart? AGARD, Neuilly-sur-Seine, France. AB - BACKGROUND: Based on physiologic considerations, observations in animal experiments and the results of a preliminary French study, there has been an aeromedical concern that repeated exposure to high sustained G-forces might have a deleterious effect on the heart. The AGARD Aerospace Medical Panel initiated a multi-national study to address the question. HYPOTHESIS: The study addressed the null hypothesis that "there is no difference in cardiac chamber dimensions, wall thickness or echocardiographic functional parameters between pilots who fly high sustained G (HSG) aircraft and pilots who fly primarily rotary wing or transport aircraft." METHODS: The study was a cross-sectional design comparing echocardiographic parameters in NATO active duty male pilots of HSG aircraft with a control group of transport and rotary wing pilots (CNTL). Some 13 NATO nations participated using a detailed protocol which included specific echocardiographic technical instructions, and procedures for collecting quantitative data on demographic variables including exercise, smoking and flying hours. Data was forwarded on a specially-designed software program to a central data registry. Careful quality control was carried out. RESULTS: Comparing data from 289 HSG pilots with 254 CNTL pilots, when corrected for the covariates, there were no differences for any of 16 echocardiographic parameters including right and left ventricular dimensions and wall thickness, aortic and left atrial dimensions, and tricuspid and mitral inflow velocities. CONCLUSIONS: The results support the null hypothesis. The conclusions are limited to the resolution of the technology employed and to the flight envelopes and +Gz exposure in the current generation of fighter aircraft. PMID- 9215465 TI - Heart rate responses to real and simulated BA Hawk MK 51 flight. AB - BACKGROUND: The effects of psychological workload on inflight heart rate were studied in five experienced (flight instructors) and five less experienced (cadets) military pilots of the Finnish Air Force (FAF). METHOD: The subjects performed the same flight mission twice; first with the BA Hawk MK 51 simulator with minimal G-forces and after that with the BA Hawk MK 51 jet trainer with Gz forces below +2. The mission included: a) 2 min rest after seating; b) take-off; c) ILS approach in the minimum weather conditions (initial, intermediate and final approach); d) landing tour (visual approach); and e) landing. The heart rates were continuously measured using a small portable recorder developed at the University of Jyvaskyla, Finland. The R-R intervals were stored and analyzed with an accuracy of 1 ms. The different phases of each flight were marked in the data by using codes given beforehand for each critical event. RESULTS: The take-off resulted in a significant increase in the heart rate from the resting levels both in the cadets and the flight instructors in both planes. In the simulator the heart rate decreased during the initial approach and slightly increased after it during the intermediate approach. Thereafter the heart rate decreased during the landing tour which seemed to be the least psychologically demanding phase of the simulated flight. The heart rate increased again during the landing but did not exceed the heart rates measured during the take-off and the ILS-approach. There were no statistical differences between the groups. In the jet trainer no decrease in the heart rate could be observed immediately after the take-off, unlike in the case of the simulated flight. The inflight heart rate increased during the final approach, decreased during the landing tour and finally increased during the landing. According to the heart rate analysis the final approach was the most loaded phase of the real flight. The changes towards the phases of final approach and landing were greater among the flight instructors. CONCLUSION: There were no statistically significant differences between the mean heart rates during the real and the simulated flight. It is suggested that the heart rate changes for most reflected the changes in cognitive workload. PMID- 9215466 TI - Activated protein C resistance as a "new" cause of deep venous thrombosis in aviators. AB - Aviators are occasionally diagnosed as suffering from deep venous thrombosis (DVT). Despite the recognition of the "Economy Class Syndrome" in the 1960's, the relationship between DVT and aviation is not clear cut. A case of DVT is described in a military navigator who was also found to be a heterozygote for activated protein C resistance. This case highlights the importance of considering all components of Virchow's triad when assessing the significance and management of DVTs. Aspects of the Economy Class Syndrome and of activated protein C resistance are discussed. PMID- 9215467 TI - Eccentric simple bone cysts of the femoral neck in adults. AB - The purpose of this communication is to describe atypical simple bone cysts of the femoral neck seen in adult patients. Two patients, aged 56 and 49, having cystic lesions which did not conform to a typical simple bone cyst, are reported. Common features including eccentric location, and thick lining tissue and sclerotic margin are not those of typical simple bone cysts seen in children. Local mechanical characteristics might be related to the atypical presentations. PMID- 9215468 TI - Epiphyseal growth arrest lines. MR findings. AB - We present two cases in which MR exams revealed unusual, low signal intensity lines in the marrow space of epiphyses. These epiphyseal lines were smooth and regular, creating a bone-in-bone appearance. These lines were much more conspicuous on MR than on radiographs, partly because of adjacent alterations in trabecular architecture. A history of prolonged immobilization during childhood in both cases suggests that these lines represent growth arrest lines persisting in the epiphyses. PMID- 9215469 TI - Superficial siderosis of the central nervous system. Its MRI manifestations. AB - Three cases of superficial siderosis of the central nervous system are reported here. Using a 1.0-T magnetic resonance (MR) unit, typical hypointense rims were observed under the brain surface on T2-weighted images. In one patient, marked atrophy of the superior cerebellar vermis and cerebellar parenchymal hyperintensity were also detected. The spinal cord was involved in two of the three patients. On T1-weighted images, hyperintense rims were demonstrated over the brain surface in two of the three patients. This finding has not been previously reported. PMID- 9215470 TI - MR of post traumatic spinal cord lesions. Unexpected improvement of hemorrhagic lesions. AB - Fifteen patients who sustained spinal cord trauma were evaluated by MR within 72 hours of injury. Nine patients had hemorrhagic and six had nonhemorrhagic traumatic spinal cord lesions. Three patients with hemorrhagic and all six patients with nonhemorrhagic lesions showed some degree of neurological improvement on follow-up examinations. In two of the three patients with hemorrhagic lesions who improved, the hemorrhage was extensive. This supports the observation that hemorrhagic lesions are not always associated with a poor clinical outcome. PMID- 9215471 TI - A second look at unenhanced spinal magnetic resonance imaging of malignant leptomeningeal disease. AB - The purpose of this study is to evaluate the use of unenhanced magnetic resonance (MR) imaging in identifying malignant leptomeningeal disease (MLD). Included in this study were fifty patients with evidence of leptomeningeal enhancement on post-gadolinium MR images and with cytological confirmation of MLD. Unenhanced, T1-weighted spin-echo (SE) MR images of the spine were analyzed for loss of cerebrospinal fluid (CSF) clarity, poor definition of the conus medullaris, thickened and clumped nerve roots, and nodules. Patterns of leptomeningeal enhancement on post-gadolinium, T1-weighted SE images were noted. Findings of MLD on unenhanced MR images were observed in 41 (85%) of 48 studies of the lumbar spine, 10 (50%) of 20 studies of the thoracic spine, and two (33%) of six studies of the cervical spine. In the lumbar spine, thickened and clumped nerve roots, poor definition of the conus medullaris, loss of CSF clarity, and nodules were observed with decreasing frequency. The signs of MLD on unenhanced images of the cervicothoracic spine included nodules and clouding of CSF. Patterns of leptomeningeal enhancement included linear, linear/nodular, nodular, enhancement of nerve roots, and stacking, with tumor filling the lumbosacral canal. Findings of MLD were present on 73% of the unenhanced MR images of the spine. Recognition of MLD on unenhanced MR images can guide the appropriate work-up and therapeutic approach. PMID- 9215472 TI - Upper extremity DVT. Correlation of MR and nuclear medicine flow imaging. AB - The purpose of this study is to determine correlation between MR and nuclear medicine flow studies in the evaluation of upper extremity deep venous thrombosis. We retrospectively reviewed MR and radionuclide venography images obtained in 10 patients with suspected upper extremity venous thrombosis. In nine cases there was complete agreement in the identification of thrombus. In one case, MR images failed to identify a thrombus in the axillary vein adjacent to a port-a-cath. Good correlation between MR and radionuclide venography images in evaluation of upper extremity DVT is present in the majority of cases. PMID- 9215473 TI - Magnetic resonance imaging of myelofibrosis. STIR and gadolinium-enhanced MR images. AB - MR images of myelofibrosis were assessed in twelve patients. Myelofibrosis showed low intensity in T1-weighted images, high intensity in STIR images, and enhancement after gadolinium injection. MR spectroscopy detected a large water resonance. MR imaging was consistent with histological findings and useful in evaluating the extracellular space in bone marrow of myelofibrosis, but it was of no value in differentiating between primary and secondary myelofibrosis. PMID- 9215474 TI - Foreign body granuloma simulating solid neoplasm on MR. AB - We report a postoperative foreign body granuloma occurring in the thyroid bed after anterior cervical fusion. The magnetic resonance (MR) imaging finding of increased signal on T2-weighted images is atypical for this type of granuloma and is thought to be due to the large proportion of cellularity in the mass histologically. In the differential diagnosis of solid postoperative masses it is important to consider foreign body granuloma, as it can mimic neoplastic disease both clinically and radiographically. PMID- 9215475 TI - Recurrent renal cell carcinoma after 45 years. AB - Late recurrence of renal cell carcinoma (RCC), arbitrarily defined as > 10 years post nephrectomy, is rare. The longest known clinical disease-free interval of 36 years was reported by Walter and Gellespie in 1960. We report a case of recurrent RCC presenting 45 years after nephrectomy. PMID- 9215476 TI - Improved sonographic visualization by fluid challenge method of renal lithiasis in the nondilated collecting system. Experience in seven cases. AB - Ultrasound is an effective means of detecting renal stones in patients with hydronephrosis. The filled urinary bladder in a normal person under hydration frequently results in the sonographic appearance of minimal or moderate hydronephrosis. For optimal visualization of stones and associated acoustic shadows, six patients with radiographically opaque renal stones were evaluated with fluid-loaded renal ultrasonography, which was compared to conventional renal sonogram in basal state. Although ultrasound in basal state can be used for detection of intrarenal lithiasis, fluid-challenged renal sonogram appears more accurate for the determination of contour, location, size, and number of small stones. In one case, this fluid-loaded ultrasound well demonstrated the tramline appearance of an isolated renal artery, which mimics stone before fluid challenge. PMID- 9215477 TI - Ovarian hyperstimulation syndrome. US and CT appearances. AB - Ovarian hyperstimulation syndrome has been reported following induction of ovulation with drug therapy for infertility. The ultrasonic and computed tomographic (CT) appearances of the ovarian cystic enlargement is described. The role of ultrasound in this condition is diagnostic; the results of which are supported by CT. PMID- 9215478 TI - Spontaneous renal hemorrhage associated with renal tumors. AB - Spontaneous ruptures of the kidney sometimes require emergency surgery, at which time the etiology for the rupture becomes evident. Because the patient with previously existing renal pathology is asymptomatic, when these ruptures do occur one should be suspect of underlying disease. We present a case and discuss the relevant aspects of such entities. PMID- 9215479 TI - MR imaging of carcinoma within urinary bladder diverticulum. AB - Cystoendoscopic examination and standard radiological techniques occasionally fail to correctly establish that a pelvic mass is due to carcinoma arising within a urinary bladder diverticulum. MR imaging in oblique planes can facilitate the diagnosis in such cases by demonstrating the neck of the diverticulum. Also, T2 weighted images allow differentiation between tumor within a diverticulum and a necrotic extravesical mass. PMID- 9215480 TI - MRI of the liver. A pictorial essay. AB - Magnetic resonance imaging (MRI) is an extremely useful modality for evaluation of the complex pathophysiology of the liver. The high degree of soft tissue contrast afforded by MRI accurately detects and characterizes both focal and diffuse abnormalities of the liver. In this article we present a pictorial review of MRI of the liver. PMID- 9215482 TI - AUDILAB: a knowledge-based quality audit simulator for testing laboratories. AB - In order to obtain an accreditation, a laboratory must be prepared to provide a point-by-point check of various activities against the chosen reference standard, both from a general point of view and in relation to details of application. This paper describes AUDILAB, a computerized simulator accessible by network, able to provide testing laboratories with realistic quality audits performed in a customized way. AUDILAB establishes a detailed list of strengths (compliance with corresponding requirements of established standards) and weaknesses (improvements needed for laboratory's accreditation). The standard used by AUDILAB is the EN 45001 "General criteria for the operation of testing laboratories". A preliminary validation has already been completed. AUDILAB became operational in September 1993. PMID- 9215481 TI - THEMPO: a knowledge-based system for therapy planning in pediatric oncology. AB - This article describes the knowledge-based system THEMPO (Therapy Management in Pediatric Oncology), which supports protocol-directed therapy planning and configuration in pediatric oncology. THEMPO provides a semantic network controlled by graph grammars to cover the different types of knowledge relevant in the domain, and offers a suite of acquisition tools for knowledge base authoring. Medical problem solvers, operating on the oncological network, reason about adequate therapeutic and diagnostic timetables for a patient. Furthermore, a corresponding patient record, also based on semantic networks and graph grammars, has been implemented to represent the course of therapy of an oncological patient. PMID- 9215483 TI - Cancellation of motion artifact in MRI due to 2D rigid translational motion. AB - A new algorithm for cancelling MRI artifact due to translational motion in the image plane is described. Unlike the conventional iterative phase retrieval algorithm, in which there is no guarantee for the convergence, a direct method for estimating the motion is proposed. In the previous approach, the motions in the readout(x-) direction and the phase encoding(y-) direction are estimated simultaneously. However, the feature of the each x- and y-directional motion is different. Based on the analysis of their features, each x- and y-directional motion is cancelled by different algorithms in two steps. First, we notice that the x-directional motion corresponds to a shift of the x-directional spectrum of the MRI signal, so the x-directional motion can be cancelled by shifting the spectrum in inverse direction. Next, the y-directional motion is cancelled using a new constraint, with which the motion component and the true image component can be separated. The algorithm is shown to be effective by simulations. PMID- 9215484 TI - Exploring a mixture of distributions using Minitab. AB - A set of macros has been developed to help the user explore the characteristics of the two-component mixture of distributions. Five distributions are included: normal; exponential; Weibull; lognormal; and uniform. Using the powerful abilities of Minitab (version 9) the user can produce graphs for the conditional and unconditional density, cumulative density, survival and hazard functions. The mean of the unconditional distribution is calculated as the weighted average of the conditional means. A technique is programmed for calculating the unconditional median which cannot be derived mathematically. A What..if..? approach can be helpful in clarifying concepts such as sufficient conditions for unimodality, identifiability of mixtures, and hazard function of mixture of distributions. PMID- 9215485 TI - The assignment of velocity profiles in finite element simulations of pulsatile flow in arteries. AB - In this paper we present a new method for the assignment of pulsatile velocity profiles as input boundary conditions in finite element models of arteries. The method is based on the implementation of the analytical solution for developed pulsatile flow in a rigid straight tube. The analytical solution provides the fluid dynamics of the region upstream from the fluid domain to be investigated by means of the finite element approach. In standard fluid dynamics finite element applications, the inlet developed velocity profiles are achieved assuming velocity boundary conditions to be easily implementable-such as flat or parabolic velocity profiles-applied to a straight tube of appropriate length. The tube is attached to the inflow section of the original fluid domain so that the flow can develop fully. The comparison between the analytical solution and the traditional numerical approach indicates that the analytical solution has some advantages over the numerical one. Moreover, the results suggest that subroutine employment allows a consistent reduction in solving time especially for complex fluid dynamic model, and significantly decreases the storage and memory requirements for computations. PMID- 9215486 TI - Theoretical analysis of the mechanisms of chronic hyperinsulinemia. AB - Steady-state insulin resistance results in a fasting hyperinsulinemia and is a common feature of type II diabetes mellitus and obesity. In this study, a systems analysis approach was used to study glucose homeostasis which is considered as the dynamic balance between glucose release by the liver and its uptake by the peripheral tissues as regulated by insulin and glucagon. A series of computer simulation studies were performed utilizing a mathematical model of glucose homeostasis. The purpose of the study was to better understand the factors which control glucose balance and to ascertain their relative importance in the development of steady state, fasting hyperinsulinemia. When peripheral cellular insulin receptors which regulate glucose uptake were reduced to 25% of normal, the steady state plasma insulin concentration showed little change from the basal level of 8 microU/ml. When insulin receptors in the liver were also reduced to 25% of normal, the steady state insulin concentration increased from the basal levels to 32 microU/ml. Reducing pancreatic alpha cell insulin receptors to 25% of normal further increased the hyperinsulinemia to 80 microU/ml. Hence, this study suggests that the liver and its release of glucose, as controlled by insulin and glucagon, plays a central role in the development of a steady-state insulin resistance and hyperinsulinemia. PMID- 9215487 TI - Umbilical artery blood gases in healthy term newborn infants. AB - OBJECTIVE: To study the distributions of pH and gas values in umbilical arterial (UA) blood of normal newborns following uncomplicated pregnancies and vaginal births. METHODS: In 681 consecutive normal term infants who were born during 1990, we examined the UA pH and blood gas values obtained immediately following delivery. Maternal inclusion criteria were defined as an uncomplicated pregnancy and a normal spontaneous vaginal delivery. Umbilical artery blood samples were collected at each birth and were evaluated for pH, carbon dioxide pressure (PaCO2), oxygen pressure (PaO2), bicarbonate, and base excess. All newborns in this study had the following inclusion criteria: singletons, no malformations, growth appropriate for gestational age (AGA), and Apgar scores of 7 or more at 1, 5 and 10 min of life. RESULTS: The lowest UA pH was 7.04 and the 10th percentile value was 7.21. The lowest UA PaO2 was 4.6 mmHg and the 10th percentile value was 10.1 mmHg. The highest UA PaCO2 was 75.4 mmHg and the 90th percentile 62 mmHg. The 10th percentile of the base deficit in the extracellular fluid was 5.9 mmol/l. CONCLUSIONS: The distributions of the UA pH and gas values of a collective of normal newborns were illustrated. PMID- 9215488 TI - Full-term birth weight and placental morphology at high and low altitude. AB - OBJECTIVE: To study the association between placental morphology and full-term birth weight at high and low altitude. SUBJECTS: Twenty normal pregnant women living permanently at high altitude (3100 m) and 20 normal pregnant women living permanently at low altitude (500 m) in Southern Saudi Arabia. METHOD: For each subject in the two groups the mean hemoglobin concentration and hematocrit values throughout pregnancy were estimated and these were used as indices for maternal hypoxia. After delivery, the birth weight of each fetus was determined together with the placental weight. Placentas were then examined histologically using sections stained by periodic acid-Schiff and hematoxylin-eosin. The mean percentages of villi with syncytial knots, cytotrophoblastic cells and fetal capillaries were determined. RESULTS: The mean hemoglobin concentration and hematocrit values were significantly greater at high altitude than at low altitude (P < 0.001 for both). The mean birth weight and placental weight were significantly greater at low altitude compared to high altitude (P < 0.025 and 0.001, respectively). The placentas from high altitude showed histological changes suggestive of placental hypoxia i.e. significant increase in the incidence of syncytial knots, cytotrophoblastic cells and fetal capillaries at high altitude compared to low altitude (P < 0.005, 0.001 and < 0.05, respectively). At both high and low altitude the incidences of syncytial knots and cytotrophoblastic cells showed positive and significant correlations with mean maternal hemoglobin (r = 0.5 and 0.6, P < 0.01 and < 0.001, respectively) and hematocrit (r = 0.5 and 0.6, P < 0.01 and 0.001, respectively) during pregnancy and negative and significant correlations with fetal birth weight (r = 0.4 and -0.6, P < 0.01 and P < 0.001, respectively). CONCLUSION: The low birth weight observed at high altitude compared to low altitude appeared to be mainly secondary to placental hypoxia resulting from maternal hypoxia which in turn was caused by high altitude hypoxia. PMID- 9215489 TI - Are Hong Kong babies getting bigger? AB - OBJECTIVE: To establish recent birth weight trends in Hong Kong. METHOD: A total of 10,512 confinements for the years 1985-86, and 7857 for the years 1995-96 were analyzed. RESULT: There was a significant increase in maternal height, weight at booking, and maternal age, whereas the body-mass index was reduced slightly (P < 0.0001). Parity increased significantly, with the percentage of parous women rising from 44.1% to 55.6% (P < 0.0001). The percentage of female infants decreased from 49.5% to 47.9%. Despite these changes there was no significant difference in mean birth weights between the two groups. When birth weight was controlled for sex, parity, maternal height and weight there was a trivial increase of 15 g over time (P = 0.01). CONCLUSION: Birth weight has reached a plateau in Hong Kong despite a continuing increase in the regions' socioeconomic status, and evidence of improved nutritional well-being. PMID- 9215490 TI - Cesarean section using the Misgav Ladach method. AB - OBJECTIVE: To stress the advantages of the Misgav Ladach method for cesarean section. STUDY DESIGN: In this study operative details and the postoperative course of 139 patients who underwent cesarean section according to the Misgav Ladach method in 1995-96 are presented. RESULTS: The Misgav Ladach method reduces operation time, time of child delivery, and time of recovery. The rates of febrile morbidity, wound infection and wound dehiscence are not affected by the new technique. CONCLUSION: Our study highlights the efficiency and safety of the Misgav Ladach method, and points out the speeded recovery, with early ambulation and resumption of drinking and eating, that makes the cesarean section delivery closer and closer to natural childbirth. PMID- 9215491 TI - Single-dose systemic oral fluconazole for the treatment of vaginal candidiasis. AB - OBJECTIVES: To evaluate the acceptance of fluconazole given in a single oral dose, for the safe, effective treatment of vaginal candidiasis. METHODS: A total of 428 patients who had a first or recurrent episode of vaginal candidiasis diagnosed clinically or by culture, were offered treatment with fluconazole by 40 primary care gynecologists who were unfamiliar with fluconazole treatment of vaginal candidiasis. The efficacy of this treatment was evaluated by both physicians and patients. RESULTS: Most of the physicians (72%) and most of the patients (69%) found the drug effective in relieving or at least alleviating the signs and symptoms of the disease. The majority of patients (83.5%) rated it better than other drugs they had received for vaginitis in the past. No recurrences were noted at the 6-week follow-up. CONCLUSIONS: Fluconazole has been found effective by physicians and patients. Both physician willingness to use it and patient compliance are satisfactory. PMID- 9215492 TI - Lipids in serum of patients with malignant ovarian neoplasms. AB - OBJECTIVE: In numerous papers, different associations of serum lipids and lipoproteins with neoplastic diseases were found. The aim of our work was to find out associations between levels of 11 serum lipids, lipoproteins and ovarian cancer. METHODS: 25 healthy women and 25 patients with ovarian cancer underwent examinations. Uni- and multivariate analysis of variance and discrimination analysis were used to analyze our results. RESULTS: The analysis demonstrates that ovarian carcinoma is associated with a significant reduction of total cholesterol and its esters in serum and in high density lipoprotein fractions compared to controls. CONCLUSION: It could be shown that using multivariate analysis of variance it is possible to find the optimal set of serum lipid parameters, containing serum esterified cholesterol, serum total cholesterol and high density lipoproteins esterified cholesterol, to differentiate healthy persons from patients affected by ovarian cancer. PMID- 9215493 TI - Radiation therapy duration influences overall survival in patients with cervical carcinoma. AB - OBJECTIVE: This article analyses the influence of treatment duration on survival in patients with invasive carcinoma of the cervix treated by radical radiation therapy. METHOD: Three hundred and sixty patients with FIGO stage IB-IIIB carcinoma of the cervix were treated in Lausanne (Switzerland) with external radiation and brachytherapy as first line therapy. Median therapy duration was 45 days. Patients were classified according to the duration of the therapies, taking 60 days (the 75th percentile) as an arbitrary cut-off. RESULTS: The 5-year survival was 61% (S.E. = 3%) for the therapy duration group of less than 60 days and 53% (S.E. = 7%) for the group of more than 60 days. In terms of univariate hazard ratio (HR), the relative difference between the two groups corresponds to a 50% increase of deaths (HR = 1.53, 95% CI = 1.03-2.28) for the longer therapy duration group (P = 0.044). In a multivariate analysis, the magnitude of estimated relative hazards for the longer therapies are confirmed though significance was reduced (HR = 1.52, 95% CI = 0.94-2.45, P = 0.084). CONCLUSION: These findings suggest that short treatment duration is a factor associated with longer survival in carcinoma of the cervix. PMID- 9215494 TI - Responses of polycystic ovarian syndrome and related variants to low-dose follicle stimulating hormone. AB - OBJECTIVE: To determine whether patients with classical polycystic ovarian syndrome (PCOS) respond differently to low-dose FSH therapy in comparison with anovulatory patients (PCOS-like) where only some features of PCOS are present. METHODS: Two groups of patients were studied. The PCOS group (25 patients, 51 cycles) and the PCOS-like group (38 patients, 57 cycles) were treated with the same protocol of purified FSH, commencing with an initial dosage of 75 IU/day and increasing by 37.5 IU/day after 14 days, where necessary. Estradiol levels and ultrasonographic evidence of follicular development were used for monitoring. RESULTS: PCOS patients required more ampules of FSH, needed more days of gonadotropin stimulation, secreted higher levels of E2 and had increased numbers of intermediate follicles compared to the PCOS-like group. CONCLUSIONS: This study demonstrated significant differences between PCOS and other PCOS-like conditions when treated with low-dose FSH. Classification of the subvariants of PCOS may have therapeutic implications. PMID- 9215495 TI - Glanzmann's thrombasthenia and puerperium. PMID- 9215496 TI - Severe primary postpartum hemorrhage. PMID- 9215497 TI - Fetal trisomy 13 distinguished by persistent fetal tachycardia. PMID- 9215498 TI - ACOG educational bulletin. Teratology. Number 236, April 1997 (replaces no. 233, February 1997). American College of Obstetricians and Gynecologists. PMID- 9215499 TI - ACOG committee opinion. Ethical issues in obstetric-gynecologic education. Number 181, April 1997. Committee on Ethics. American College of Obstetricians and Gynecologists. PMID- 9215500 TI - ACOG criteria set. Ambulatory care criteria set: hormone replacement therapy. Number 23, April 1997. Committee on Quality Assessment. American College of Obstetricians and Gynecologists. PMID- 9215501 TI - FIGO News. Report of the FIGO Committee for the Study of Ethical Aspects of Human Reproduction. International Federation of Gynecologists and Obstetrics. PMID- 9215502 TI - Body dissatisfaction among Hispanic and Asian-American girls. PMID- 9215503 TI - They practice what we teach: more on adolescent health care in pediatric practice. PMID- 9215504 TI - Ethnic differences in childhood and adolescent sexual abuse and teenage pregnancy. AB - PURPOSE: This study examined ethnic differences in childhood and adolescent sexual abuse and the effect on teenage pregnancy rates. METHODS: A 20-page questionnaire elicited information about women's sexual and pregnancy history, high-risk behaviors, and sexual abuse, based a modified version of the Koss and Oros Sexual Experiences Survey. Over 1,900 women between 18 and 22 years old were recruited at 44 urban and rural sites. Women representing four ethnic groups completed an English or Spanish version of the questionnaire. RESULTS: Almost 36% of the women reported sex abuse before age 18 years of age, and more than 26% were pregnant before reaching 18 years old (teenage pregnancy). Compared with their nonabused peers, twice as many women who were coerced into sex or raped had a teenage pregnancy. Minority group teens were more likely than Anglos to have a teenage pregnancy and to have been coerced into having sex, rather than raped, prior to teenage pregnancy. CONCLUSIONS: Over one-third of pregnant teenagers in this study have been sexually abused, usually involving sexual intercourse, prior to becoming pregnant. Coercive sexual abuse is more likely to contribute to teenage pregnancy among minority group teens, whereas rape is more likely to contribute to a teenage pregnancy among Anglos. PMID- 9215505 TI - Efficacy of a dating violence prevention program on attitudes justifying aggression. AB - OBJECTIVES: The purpose of this pilot study was to evaluate a five-session dating violence prevention curriculum in terms of its effect on attitudes justifying the use of dating violence. METHODS: The curriculum was implemented in all health classes in a Long Island, New York, school. A total of 193 students participated (boys, n = 106; girls, n = 87). A quasi-experimental design was used to evaluate change in attitudes justifying dating violence, with health classes randomly assigned to the treatment or no-treatment conditions. RESULTS: Pre- to postprogram assessments indicated that there were significant decreases in overall attitudes justifying the use of dating violence as a means to resolve conflict among students exposed to the curriculum material, whereas those who were not exposed did not show attitude change from pre- to postprogram evaluation. CONCLUSIONS: The curriculum shows promise as an effective tool for changing attitudes condoning dating violence. Future research is needed to determine whether the observed attitude change is also linked to reduction in aggressive behaviors. PMID- 9215506 TI - Improving detection of violence among pregnant adolescents. AB - PURPOSE: The purpose of this study was to determine whether a systematic assessment protocol could increase reporting of violence among pregnant adolescents compared with a routine prenatal assessment. This study also sought to examine issues related to violence assessment among maternity care coordinators. METHODS: The Maternity Care Coordination (MCC) program in a health department prenatal clinic in North Carolina routinely screened all clients for violence at their first visit. This assessment was not standardized. In 1994, the MCC program implemented a systematic violence assessment protocol for all adolescents (n = 117). The protocol assessed violence at three points during pregnancy by asking one direct question: "Have you been hit, slapped, kicked, or hurt during this pregnancy?" To examine the effectiveness of the system, we retrospectively reviewed the 1993 MCC records in which the coordinators routinely screened clients for violence (n = 129). To examine issues related to screening, we conducted in-depth interviews with the maternity care coordinators. RESULTS: The routine pre-intervention assessment indicated that 5.4% of adolescents 12-19 years of age reported prenatal violence. The systematic assessment protocol resulted in a significant increase in reported violence from 5.4% to 16.2% (odds ratio = 2.9, 95% confidence interval = 1.6, 5.6, adjusted for race). Maternity care coordinators identified five factors related to increased reporting using the standardized protocol: (a) written protocol and data collection form; (b) asking direct, specific questions; (c) not labeling the victim; (d) not naming the perpetrator; and (e) conducting multiple assessments. CONCLUSIONS: Multiple, direct, systematic assessments throughout prenatal care resulted in increased reporting of prenatal violence among adolescents compared to single, routine, nonstructured assessments. PMID- 9215507 TI - Patterns of health risk behaviors for chronic disease: a comparison between adolescent and adult American Indians living on or near reservations in Montana. AB - PURPOSE: To compare the chronic disease health risk behavior patterns of adolescents and adults among American Indians living on or near reservations in Montana. METHODS: We analyzed data from the 1993 Youth Risk Behavior Survey of American Indians in Grades 9-12 living on or near Montana reservations. Risk factors included tobacco use, low physical activity, attempted weight loss, and low consumption of fruits, vegetables, and green salad. Similar data were analyzed from a 1994 Behavioral Risk Factor Survey of American Indian adults living on or near reservations in Montana. RESULTS: The prevalence of most adolescent health risk behaviors was high, especially cigarette smoking (45% for males, 57% for females), smokeless tobacco use (44% for males, 30% for females), and infrequent consumption of salad or vegetables (59-76%). With the exception of daily cigarette smoking and inadequate fruit consumption among adolescents of both genders and physical inactivity among adolescent males, the prevalence of chronic disease health risk behaviors among adolescents was similar to or higher than the prevalence of the same risk behaviors among adults. CONCLUSIONS: Many health risk behaviors for chronic diseases are common by the time this group of American Indians in Montana has reached adolescence. Possible reasons may include modeling of familial behaviors, peer pressure, advertising, or age cohort effects. If these risk behavior patterns continue into adulthood, morbidity and mortality from chronic diseases are likely to remain high. Substantial efforts are needed to prevent or reduce health risk behaviors among adolescents and adults in this population. PMID- 9215508 TI - Adolescent knowledge, values, and coping strategies: implications for health in sub-Saharan Africa. AB - PURPOSE: The purposes of this study were to investigate the experiences and knowledge of adolescents living in an urban center in Kenya and to understand how the decisions they make affect their physical and psychological health. METHODS: A sample of 216 adolescents was drawn and they met weekly in small groups with trained facilitators for a period of 6 months. A research team monitored the developments of the groups and the topics they discussed. The findings were corroborated by the adolescents. RESULTS: It was found that adolescents were primarily concerned with developing a coherent and consistent set of personal values which would govern their behavior. Unfortunately, they could not always achieve those values and they resorted to dysfunctional coping strategies which were injurious to their health. CONCLUSIONS: The role of the adolescent in developing countries is complex and poorly defined. In a period of unprecedented change, an urgent and comprehensive review is necessary by all sections of society if the health of this group is to improve. PMID- 9215509 TI - Gender and social differences in adolescent sexuality and reproduction in Nicaragua. AB - PURPOSE: The aim of this research was to study gender and social differences in adolescent sexuality and reproduction, as reflected in age at first intercourse and age at first pregnancy, as a basis for future interventions in the municipality of Leon, Nicaragua. METHODS: In a community-based cross-sectional study including 7789 households, all women aged 15-49 years (n = 10,867) were interviewed about socioeconomic, sexual, and reproductive issues. A random subsample of men (n = 388) and women (n = 413) aged 15-49 years was interviewed in more detail about sexual patterns and reproduction. RESULTS: The median age at first intercourse for women and men was 17.8 and 16.2 years, respectively. Women's average latency period to end of first pregnancy was 22 months. There was a significant tendency to start active sexual life later among today's girls aged 15-20 years, compared to the groups 21-27, 28-35, and 36-49 years old. A similar trend was found in age at first pregnancy. These secular trends were not found among men. Age at first pregnancy for current adolescents was lower among those having less formal education. CONCLUSIONS: The short latency period between first sexual intercourse and end of first pregnancy, probably reflecting lack of access to counseling and contraception, is worrying in light of the growing sexually transmitted disease/human immunodeficiency virus threat. The secular trend of later start of reproduction, however, is a positive sign which partly may be an effect of increasing education in the Nicaraguan society. PMID- 9215510 TI - Adolescent dimenhydrinate abuse: resurgence of an old problem. PMID- 9215511 TI - Crohn disease of the esophagus in an adolescent. PMID- 9215512 TI - Bibliography of journal articles (January to June 1995). PMID- 9215514 TI - Women married to men with myocardial infarction are at increased risk of coronary heart disease. AB - AIM: To measure the prevalence of coronary heart disease (CHD) risk factors in the female partners of men with acute myocardial infarction (AMI). METHOD: Consecutive incident cases of men under 65 years of age with AMI surviving to 3 months, their female partners and female healthy controls matched for age and marital status drawn from the general population were investigated. RESULTS: One hundred and seventeen cases of AMI in men under 65 years of age and 89 female partners were identified; 133 age- and sex-matched controls were examined for CHD risk factors. Cigarette smoking was more common among the younger partners (25-44 years of age) compared with controls. A body mass index > 28 kg/m2, systolic blood pressure > 150 mmHg, diastolic blood pressure > 90 mmHg and cholesterol > 6.5 mmol/l were all significantly more common in partners compared with controls. In a logistic regression of age, smoking habit, blood pressure, cholesterol and body mass index, based on 89 female partners and 132 controls with complete data, body mass index > 28 kg/m2 (odds ratio 2, 17, 95% CI 1.11-4.23) and cholesterol > 6.5 mmol/l (odds ratio 2.21, 95% CI 1.08-4.49) were both significantly more common in the female partners compared with controls. CONCLUSIONS: Women married to men with AMI have a higher frequency of CHD risk factors compared with married women in the general population, consistent with a shared family lifestyle putting both adults at higher risk of CHD. Screening blood relatives in families prematurely affected by CHD is widely advocated; such screening should include partners. PMID- 9215513 TI - Comparing two strategies to modify dietary behavior and serum cholesterol. AB - AIM: To test the hypothesis that a strategy including cholesterol screening and dietary education is more effective than dietary education alone in changing dietary behavior and serum cholesterol levels. METHODS: Individuals at four worksites were enrolled in a randomized trial with a 'full intervention' condition in which subjects were told their serum cholesterol value and also received a dietary change kit (n = 236), and a 'partial intervention' condition in which subjects received the same dietary change kit, but were not told their serum cholesterol value (n = 284). Individuals (n = 115) in two worksites served as a nonrandomized 'untreated control group'. Subjects were tested for serum cholesterol and completed a questionnaire at baseline, and 3 and 6 months later. RESULTS: Dietary changes occurred in seven of nine categories in individuals subjects to the full and partial interventions but in only one of nine categories in those studied in the control condition. Mean dietary intake differed between the full and partial intervention conditions for only three of nine dietary categories. Cholesterol level dropped in the full, partial and control conditions by 4.9, 3.9 and 9.6%, respectively. CONCLUSIONS: Dietary education has favorable effects on the dietary behaviors of individuals. Being told one's cholesterol level at the outset of this educational intervention has little effect on dietary change. PMID- 9215515 TI - Premature coronary heart disease: the influence of positive family history on platelet activity in vivo in children and adolescents (family study). AB - BACKGROUND: Hypercoagulative disturbances are an integral part of atherosclerosis. The estimation of beta-thromboglobulin (BTG) concentrations in the offspring of parents suffering from premature coronary heart disease (CHD) makes it possible to evaluate platelet activity in vivo in this high-risk group. METHODS: The study included 292 individuals. BTG concentrations in platelet-poor plasma were estimated in 90 offspring (60 boys and 30 girls, aged 7-18 years) of fathers who had experienced a premature infarct (aged 45 years or younger) and in their parents (the main group, n = 185). All the participants were tested under their usual living conditions. Fifty-nine healthy children and adolescents of the same age with no family history of vascular events, diabetes or hypertension, together with their parents, formed the control group (n = 107). RESULTS: The mean BTG level in the main group was significantly elevated (86.04 +/- 8.14 ng/ml in the boys and 75.57 +/- 8.55 ng/ml in the girls), when compared with the values for their respective controls (49.30 +/- 3.54 ng/ml and 52.56 +/- 3.42 ng/ml, P < 0.001 and P < 0.005). High BTG levels (higher than the 95th percentile in the controls) were observed in 33.3 +/- 6.1% of the boys and in 23.3 +/- 7.7% of the girls in the main group. Twelve months later, a follow-up study of 16 offspring from the main group and 12 from the control group demonstrated that the increase in BTG levels in children and adolescents with infarction heredity over those observed in children without such a background was sustained (85.86 +/- 16.88 ng/ml compared with 75.90 +/- 14.02 ng/ml, P > 0.5). Significantly raised levels of BTG were also detected in the wives of men who had experienced premature infarcts, when these women were compared with respective controls (86.13 +/- 9.85 ng/ml and 45.04 +/- 4.82 ng/ml, P < 0.0001). CONCLUSION: Spontaneous platelet activation in vivo under normal living conditions was observed in a considerable proportion of children and adolescents with a family history of premature CHD. This activation is expected to constitute an endogenous risk-factor and is probably one of the mechanisms by which atherosclerosis advances before it becomes clinically apparent. PMID- 9215516 TI - The effect of antioxidant vitamin supplementation on traditional cardiovascular risk factors. AB - BACKGROUND: Evidence from observational epidemiologic studies has indicated that antioxidants consumed through the diet or as dietary supplements lower the risk of developing atherosclerotic cardiovascular disease. Evidence suggesting that the major mechanism for the protective effect of antioxidants is mediated through decreased oxidation of lipids, particularly low-density lipoprotein (LDL) cholesterol is accumulating. Other evidence, however, suggests that antioxidants may influence traditional modifiable cardiovascular risk factors such as the blood pressure and serum lipids favorably. The purpose of this study was to determine the effect of antioxidant vitamin supplementation on modifiable risk factors for atherosclerotic cardiovascular disease. DESIGN: A randomized, placebo controlled, clinical trial of antioxidant vitamin supplementation, conducted at a single community-based clinical research center. METHODS: We assigned 297 retired teachers who were members of the Maryland Retired Teachers Association randomly to 2-4 months of dietary supplementation with placebo or combined antioxidant vitamin capsules providing 400 IU/day vitamin E, 500 mg/day vitamin C, and 6 mg/day beta-carotene. The outcome measures were the blood pressure, fasting serum total cholesterol, high-density lipoprotein cholesterol, LDL cholesterol, and fasting glucose. RESULTS: After 2-4 months of supplementation the combined antioxidant supplement had had no significant effect on the systolic and diastolic blood pressures, fasting serum lipids (total cholesterol, high-density lipoprotein cholesterol, and LDL cholesterol) and fasting glucose, with unadjusted and adjusted analyses. CONCLUSION: Data from this trial suggest that the protective effect from antioxidant vitamin supplementation, if there is one, likely results from mechanisms other than modification of traditionally modifiable cardiovascular risk factors. PMID- 9215517 TI - Weight changes and risk of ischaemic heart disease for middle aged and elderly men. An 8-year follow-up in the Copenhagen Male study. AB - BACKGROUND: Weight gain and weight loss are determined by a complex interplay of health, lifestyle and genetic factors. There is controversy concerning whether weight changes are associated with an increased risk of ischaemic heart disease (IHD). METHODS: The Copenhageri Male Study was initiated in 1970/1971. This paper presents the results of a prospective study using a baseline comprising survivors examined 15 years later in 1985/1986 including 2903 men (aged 53-74 years) without cardiovascular disease. They were classified according to their weight change from 1970/1971 to 1985/1986. RESULTS: There were no differences among the incidences of IHD during the period 1985/1986-1993 for men who had lost weight (> 5% weight loss), men whose weight had remained constant (within +/-5%) and men who had gained weight (> 5% weight gain); the incidence rates were 7.8, 8.4 and 7.6%, respectively, NS. Adjustment for age and confounders including disease history had no influence on this result. Only for men who had already been overweight at the time of initial study in 1970/1971 (body mass index > 28 kg/m2) was there a slightly increased risk of IHD among those who gained weight. Men in this group who gained weight were characterized by a significantly (P < 0.05) larger tendency to have the Lewis phenotype Le(a-b-), a genetic marker previously shown to be associated with an increased risk of IHD mortality. With respect to death from all causes, only those who lost weight had a significantly increased risk compared with that of the constant weight group; the incidence rates were 25.0 and 12.6%, respectively, age-adjusted P < 0.001. The incidence rate of IHD among those who gained weight was 11.8%, NS. CONCLUSION: The results suggest that an increase or decrease in weight from middle age to old age is of little clinical importance in the prediction of IHD among men without cardiovascular disease. PMID- 9215518 TI - Sex differences in age at onset of symptoms in patients with hypertrophic cardiomyopathy. AB - BACKGROUND: We hypothesized that, in hypertrophic cardiomyopathy, endogenous estrogen may delay the development of symptoms in females as compared with males by several cardiovascular protective mechanisms. We examined this by assessing the sex distribution in relation to age at onset of symptoms in patients with hypertrophic cardiomyopathy. METHODS: The study population of 94 patients (55 men and 39 women) was subdivided into four groups according to age at onset of symptoms; < 15 years, 15-29 years, 30-39 years and > 40 years. In the first group, comprising the youngest patients, all the girls were pre-menarche. The age of 40 years was chosen as the lower limit for the last group because of the possible influence of pre- and perimenopausal decrease in estrogen levels on the onset of symptoms in women in their fifth decade of life. RESULTS: In general, females with hypertrophic cardiomyopathy developed symptoms later than did males (31.3 +/- 11.9 compared with 26.7 +/- 9.9 years; P < 0.05). Among adolescents, similar percentages of girls and boys developed the condition. Among young adult patients (between 15 and 29 years of age), it was predominantly men who developed symptoms. Men predominated, albeit insignificantly, among patients with onset of symptoms in the fourth decade of life, but there was a significant dominance of women over men in the group who became symptomatic after 40 years of age. CONCLUSION: Delayed onset of symptoms in women with hypertrophic cardiomyopathy compared with men resulted in nearly one-third of women remaining asymptomatic until 40 years of age, symptoms developing in the fifth decade of life. PMID- 9215519 TI - Association between pre-PTCA blood haemoglobin concentration and the risk of symptomatic restenosis after successful PTCA of primary coronary stenoses. AB - BACKGROUND: Thrombus formation at the dilation site has been suggested to initiate the restenosis process after percutaneous transluminal coronary angioplasty (PTCA). High haemoglobin concentrations may predispose to thrombus formation by increasing blood viscosity, slowing coronary blood flow and increasing thrombocyte adhesion. METHODS: Pre-PTCA blood haemoglobin concentrations (Hb) in 44 patients with symptomatic restenosis > or = 50% of the vessel diameter (Group A) were compared with Hb in the remaining 215 patients in a consecutive study population (Group B). RESULTS: Median Hb (range) was 149 (119 164) g/l in Group A and 142 (117-164) g/l in Group B, P = 0.004. Odds ratio (95% CI) for symptomatic restenosis was 3.22 (1.62-6.42) when Hb was dichotomised according to the median in the entire material. Hb, but not sex was a significant risk factor in multivariate analysis. CONCLUSION: Hb is a hitherto not recognized factor associated with the risk of symptomatic restenosis after PTCA and may be a link coupling male sex with increased risk of restenosis. PMID- 9215520 TI - A randomized trial of cardiovascular risk factor reduction: patterns of attrition after randomization and during follow-up. AB - BACKGROUND: The Preventive Cardiology Center compared two intensities of behavior modification for cardiovascular disease (CVD) risk factors in persons at risk and their families. Characteristics of drop-outs both before and after intervention were compared with subjects who completed the 6-month trial. METHODS: A total of 333 individuals of all ages were enrolled in the study and randomly assigned by family to a single-session ('skills' group-low-intensity) or a five-session ('practice' group-high intensity) intervention. Baseline and follow-up assessments included a personal and family health questionnaire, nutritional intake survey, and clinic visit to obtain blood pressure, lipids, and height and weight data. RESULTS: Two hundred and forty adults over 18 years of age were randomly assigned to one of the two intervention groups. Of these, 68 subjects (28.3%) did not participate in the intervention. Multivariate analysis revealed that these 'early drop-outs' were significantly more likely to be non-white and to have had a lower LDL cholesterol. Of the 172 subjects attending the intervention, 70 (40.7%) did not attend the 6-month follow-up ('late drop-outs'). Multivariate analysis revealed that, compared with follow-up attendees, non attendees were significantly more likely to be white and to be current smokers. CONCLUSIONS: Both early and late drop-outs in a randomized trial of CVD risk reduction were significantly different than continuing participants in several key factors. These differences suggest the use of caution in both interpreting and making generalizations about behavioral trials of risk factor reduction when attrition is high. PMID- 9215521 TI - Bibliography current world literature. PMID- 9215522 TI - Runt homology domain proteins in osteoblast differentiation: AML3/CBFA1 is a major component of a bone-specific complex. AB - The AML/CBFA family of runt homology domain (rhd) transcription factors regulates expression of mammalian genes of the hematopoietic lineage. AML1, AML2 and AML3 are the three AML genes identified to date which influence myeloid cell growth and differentiation. Recently AML-related proteins were identified in an osteoblast-specific promoter binding complex that functionally modulates bone restricted transcription of the osteocalcin gene. In the present study we demonstrate that in primary rat osteoblasts AML-3 is the AML family member present in the osteoblast-specific complex. Antibody specific for AML-3 completely supershifts this complex, in contrast to antibodies with specificity for AML-1 or AML-2, AML-3 is present as a single 5.4 kb transcript in bone tissues. To establish the functional involvement of AML factors in osteoblast differentiation, we pursued antisense strategies to alter expression of rhd genes. Treatment of osteoblast cultures with rhd antisense oligonucleotides significantly decreased three parameters which are linked to differentiation of normal diploid osteoblasts: the representation of alkaline phosphatase-positive cells, osteocalcin production, and the formation of mineralized nodules. Our findings indicate that AML-3 is a key transcription factor in bone cells and that the activity of rhd proteins is required for completion of osteoblast differentiation. PMID- 9215523 TI - Nuclear matrix proteins in well and poorly differentiated human breast cancer cell lines. AB - The nuclear matrix, besides providing the structural support of the nucleus, is involved in various cellular functions of the nucleus. Nuclear matrix proteins (NMPs), which are both tissue- and cell type-specific, are altered with transformation and state of differentiation. Furthermore, NMPs have been identified as informative markers of disease states. Here, the NMP profiles from human breast cancer cell lines and breast tumours were analyzed using two dimension gel electrophoresis. We identified NMPs that are associated with well and poorly differentiated human breast cancer cells in vitro and in vivo. Five NMPs (NMBC 1-5) were found to be exclusive for well-differentiated human breast cancer cells, while one NMP (NMBC-6) was found to be present only in poorly differentiated human breast cancer cells. The identification of these proteins suggests the potential use of nuclear matrix proteins as prognostic indicators. PMID- 9215524 TI - Heat stress induces tyrosine phosphorylation/activation of kinase FA/GSK-3 alpha (a human carcinoma dedifferentiation modulator) in A431 cells. AB - Exposure of A431 cells to a rapid temperature increase from 37 degrees to 46 degrees C could induce an increased expression (approximately 200% of control) and tyrosine phosphorylation/activation (approximately 300% of control) of protein kinase FA/ glycogen synthase kinase-3 alpha (kinase FA/GSK-3 alpha) in a time-dependent manner, as demonstrated by an anti-kinase FA/GSK-3 alpha immunoprecipitate kinase assay and by immunoblotting analysis with anti-kinase FA/GSK-3 alpha and anti-phosphotyrosine antibodies. The heat induction on the increased expression of kinase FA/GSK-3 alpha could be blocked by actinomycin D but not by genistein. In contrast, the heat induction on tyrosine phosphorylation/activation of kinase FA/GSK-3 alpha could be blocked by genistein or protein tyrosine phosphatase, indicating that heat stress induces a dual control mechanism, namely, protein expression and subsequent tyrosine phosphorylation to cause cellular activation of kinase FA/GSK-3 alpha. Taken together, the results provide initial evidence that kinase FA/GSK-3 alpha represents a newly described heat stress-inducible protein subjected to tyrosine phosphorylation/activation, representing a new mode of signal transduction for the regulation of this human carcinoma dedifferentiation modulator and a new mode of heat induction on cascade activation of a protein kinase. PMID- 9215525 TI - Uncoupling of p21 induction and MyoD activation results in the failure of irreversible cell cycle arrest in doxorubicin-treated myocytes. AB - Doxorubicin (Dox, Adriamicin), a potent broad spectrum anthracycline anticancer drug, selectively inhibits muscle specific gene expression in cardiac cells in vivo and prevents terminal differentiation of skeletal muscle cells in vitro. By inducing the expression of the helix-loop-helix (HLH) transcriptional inhibitor ld2, Dox represses the myogenic function of the MyoD family of muscle regulatory factors (MRFs). In many cell types, terminal differentiation is coupled to an irreversible exit from the cell cycle and MyoD plays a critical role in the permanent cell cycle arrest of differentiating myocytes by upregulating the cyclin dependent kinase inhibitor (cdki) p21. Here, we correlate Dox effects on cell cycle with changes of E2F/DP complexes and activity in differentiating C2C12 myocytes. In Dox-treated quiescent myoblasts, which fail to differentiate into myotubes under permissive culture conditions, serum re-stimulation induces cyclin/cdk re-association on the E2F/DP complexes and this correlates with an evident increase in E2F/DP driven transcription and re-entry of myoblasts into the cell cycle. Despite Dox ability to activate the DNA-damage dependent p53/p21 pathway, when induced in the absence of MyoD or other MRFs, p21 fails to maintain the postmitotic state in Dox-treated myocytes induced to differentiate. Thus, uncoupling p21 induction and MyoD activity results in a serum-reversible cell cycle arrest, indicating that MRF specific activation of cdki(s) is required for permanent cell cycle arrest in differentiating muscle cells. PMID- 9215526 TI - ADPribosylation reaction by free ADPribose in Sulfolobus solfataricus, a thermophilic archaeon. AB - In the archaeon Sulfolobus solfataricus, protein ADPribosylation by free ADPribose was demonstrated by testing both [adenine-14C(U)]ADPR and [adenine 14C(U)]NAD as substrates. The occurrence of this process was shown by using specific experimental conditions. Increasing the incubation time and lowering the pH of the reaction mixture enhanced the protein glycation by free ADPribose. At pH 7.5 and 10 min incubation, the incorporation of free ADPribose into proteins was highly reduced. Under these conditions, the autoradiographic pattern showed that, among the targets of ADPribose electrophoresed after incubation with 32P NAD, the proteins modified by free 32P-ADPribose mostly corresponded to high molecular mass components. Among the compounds known to inhibit the eukaryotic poly-ADPribose polymerase, only ZnCl2 highly reduced the ADPribose incorporation from NAD into the ammonium sulphate precipitate. A 20% inhibition was measured in the presence of nicotinamide or 3-aminobenzamide. No inhibition was observed replacing NAD with ADPR as substrate. PMID- 9215527 TI - Studies on the function of rho A protein in cardiac myofibrillogenesis. AB - The aim of this study was to provide morphological evidence for the presence of rho A protein in developing cardiomyocytes and to investigate its possible role in myofibrillogenesis. Immunostaining with a monoclonal anti-rho antibody gave a diffuse pattern in the cytosol of cultured cardiomyocytes. Introduction of C3 exoenzyme into the cells by electroporation was used to inactivate rho A protein by ADP-ribosylation. An immunostaining with anti-vinculin, anti-talin, and anti integrin antibodies showed the focal adhesions in electroporation control cardiomyocytes to be evenly distributed in the ventral sarcolemma; the costameric structure was also detected using these antibodies. In contrast, in C3 exoenzyme treated cells, focal adhesions were disassembled and costamere were absent; in addition, beta-actin-positive, non-striated fibrils were lost and assembly of M protein, titin, and alpha-actinin into myofibrils was poor, as shown by diffuse and filamentous staining pattern. C3 exoenzyme treatment had a less marked effect on mature cardiomyocytes than on immature cells; in this case, cells became distorted and few myofibrils were seen. The intensity of anti-phosphotyrosine antibody staining of the focal adhesion was also decreased or diffuse in C3 exoenzyme-treated cardiomyocytes, suggesting dephosphorylation of focal adhesion components. We therefore conclude that small G protein rho A plays an important role in myofibril assembly in cardiomyocytes. PMID- 9215528 TI - Calpain activation in shear-induced platelet aggregation. AB - Fluid shear stress has been known to activate platelet reaction such as aggregation, but the exact mechanism of shear-induced platelet aggregation (SIPA) has not been fully understood. Calpain, an intracellular calcium-activated cysteine protease, is abundant in platelets and is considered to be activated and involved in the proteolytic processes during platelet activation. A possible activation of calpain in SIPA was investigated, employing a newly developed aggregometer and specific monoclonal antibodies to detect activation of calpain. When a shear stress gradient varying between 6 and 108 dyn/cm2 was applied to platelets, activation of mu-calpain was observed only in high-shear-stressed platelets, resulting in the proteolysis of talin. At 1 min after the onset of constant high shear stress of 108 dyn/cm2, mu-calpain activation and proteolysis of talin were detected and increased in a time-dependent manner. Constant shear stress more than 50 dyn/cm2, applied for 5 min, caused mu-calpain activation and proteolysis of talin, which were increased in a shear-force-dependent manner. Calpeptin, a calpain-specific peptide antagonist, caused the complete inhibition of both mu-calpain activation and proteolysis of talin, while SIPA profiles with calpeptin showed almost no change compared to those without calpeptin. These results suggest the possibility of calpain involvement in late phases of shear induced platelet activation such as cytoskeletal reorganization. PMID- 9215529 TI - Selective effects of hydrocortisone on intestinal lipoprotein and apolipoprotein synthesis in the human fetus. AB - Studies employing human fetal intestine have yielded much interesting information on the role of polarized enterocytes in fat absorption and transport. Using the organ culture model, we examined the influence of hydrocortisone on the synthesis and secretion of lipids and lipoproteins. Human jejunal explants were cultured for 5 days at 37 degrees C in serum-free medium containing either [14C]-oleic acid or [14C]-acetate, alone or supplemented with hydrocortisone (25 or 50 ng/ml). The uptake of [14C]-oleic acid was associated with the production of triglycerides, phospholipids, and cholesteryl esters, which were all affected by hydrocortisone. This hormonal agent (50 micrograms) led to the marked reduction of secreted triglycerides (43%, P < 0.01), phospholipids (39%, P < 0.01), and cholesteryl esters (36%, P < 0.05) without altering the characteristic distribution of tissue and medium lipid classes. Similarly, hydrocortisone significantly (P < 0.01) decreased (approximately 60%) the incorporation of [14C] acetate into secreted free and esterified cholesterol in the medium. With [14C] oleic acid as a precursor, hydrocortisone significantly diminished the delivery of chylomicrons and very low density lipoproteins to the medium while consistently enhancing the secretion of high density lipoproteins. In parallel, [35S]-methionine pulse-labeling of jejunal explants revealed the concomitant inhibitory effect of hydrocortisone on apo B-100 synthesis and hydrocortisone's stimulatory effect on apo B-48 and apo A-1. These studies suggest that glucocorticoids play a critical role in lipoprotein processing during intestinal development. PMID- 9215531 TI - Deletion analysis of ors12, a centromeric, early activated, mammalian origin of DNA replication. AB - We have generated a panel of deletion mutants of ors12 (812-bp), a mammalian origin of DNA replication previously isolated by nascent strand extrusion from early replicating African Green monkey (CV-1) DNA. The deletion mutants were tested for their replication activity in vivo by the bromodeoxyuridine substitution assay, after transfection into HeLa cells, and in vitro by the Dpnl resistance assay, using extracts from HeLa cells. We identified a 215-bp internal fragment as essential for the autonomous replication activity of ors12. When subcloned into the vector pML2 and similarly tested, this subfragment was capable of autonomous replication in vivo and in vitro. Several repeated sequence motifs are present in this 215-bp fragment, such as TGGG(A) and G(A)AG (repeated four times each); TTTC, AGG, and CTTA (repeated 3 times each); the motifs CACACA and CTCTCT, and two imperfect inverted repeats. 22 and 16 bp long, respectively. The overall sequence of the 215-bp fragment is G/C-rich (50.2%), by comparison to the 186-bp (33.5% G/C-rich) minimal sequence required for the autonomous replication activity of ors8, another functional ors that was similarly isolated and characterized. PMID- 9215530 TI - Growth factor regulation of insulin-like growth factor binding protein-6 expression in osteoblasts. AB - Previously we have shown that transforming growth factor beta (TGF beta) 1, basic fibroblast growth factor (FGF), and platelet-derived growth factor (PDGF) BB inhibit the synthesis of insulin-like growth factor (IGF) II, but their effects on IGF binding protein (IGFBP)-6 in osteoblast cultures are not known. IGFBP-6 binds IGF II with high affinity and prevents IGF II-mediated effects, so that a possible mode of regulating the IGF II available to bone cells would be by changing the levels of IGFBP-6. To enhance our understanding of the actions of growth factors on the IGF II axis in bone, we tested the effects of TGF beta 1, basic FGF, PDGF BB, IGF I, and IGF II on the expression of IGFBP-6 in cultures of osteoblast-enriched cells from 22 day fetal rat calvariae (Ob cells). Treatment of Ob cells with TGF beta 1 caused a time- and dose-dependent decrease in IGFBP-6 mRNA levels, as determined by Northern blot analysis. The effect was maximal after 48 h and observed with TGF beta 1 concentrations of 0.04 nM and higher. TGF beta 1 also decreased IGFBP-6 polypeptide levels in the medium, as determined by Western immunoblot analysis. Cycloheximide at 3.6 microM decreased IGFBP-6 transcripts and prevented the effect of TGF beta 1. The decay of IGFBP-6 mRNA in transcriptionally arrested Ob cells was not modified by TGF beta 1. In addition, TGF beta 1 decreased the rates of IGFBP-6 transcription as determined by a nuclear run-on assay. In contrast, basic FGF, PDGF BB, IGF I, and IGF II did not change IGFBP-6 mRNA levels in Ob cells. In conclusion, TGF beta 1 inhibits IGFBP 6 expression in Ob cells by transcriptional mechanisms. Since IGFBP-6 binds IGF II and prevents its effects on bone cells, decreased synthesis of IGFBP-6 induced by TGF beta 1 could be a local feedback mechanism to increase the amount of IGF II available in the bone microenvironment. PMID- 9215532 TI - Intracellular distribution of heat-induced stress glycoproteins. AB - Cellular heat stress results in elevated heat-shock protein (HSP) synthesis and in thermotolerance development. Recently, we demonstrated that protein glycosylation is also an integral part of the stress response with the identification of two major stress glycoproteins, GP50, associated with thermotolerance, and P-SG67, the "prompt" stress glycoprotein induced immediately during acute heat stress. In the present study, we characterized the subcellular location and redistribution of these proteins during the cellular injury and recovery phase. In unheated and heated CHO cells, both stress glycoproteins were present in each subcellular fraction isolated by differential centrifugation. However, the subcellular redistribution in the course of cellular recovery after heat stress was specific for each stress glycoprotein. GP50 was present in all subcellular fractions before heat stress, but showed relatively little redistribution after heat stress. By 24 h of recovery following stress, GP50 showed partial depletion from lysosomes and microsomes, and was mainly present in the mitochondria. Glycosylated P-SG67 was redistributed in a more complex fashion. It was seen predominantly in the lysosomes and microsomes immediately following heat-stress, but after 6 h of recovery following heat stress, it largely disappeared from the microsomes and was present mainly in the cytosol. By 24 h of recovery following heat stress, it was found predominantly in the nucleus rich fraction and mitochondria. The localization of GP50 and P-SG67 by subcellular fractionation is consistent with immunolocalization studies and contrasts with the translocation of HSP70 after heat stress from cytosol to nuclei and nucleoli. These results reflect a characteristic distribution for each stress glycoprotein; their presence in virtually all subcellular fractions suggests multifunctional roles for the various stress glycoproteins in the cellular heat stress response. PMID- 9215533 TI - Partial purification of HLAMP-1 provides direct evidence for the multicomponent nature of the particulate matrix associated with cardiac mesenchyme formation. AB - H-LAMP-1 is a 283 kDa protein that is involved in the transformation of endothelial cells into mesenchyme within the AV canal and proximal outflow tract of the heart. This protein is part of the particulate matrix that has been suggested to be composed of multicomponent complexes that have been termed cardiac adherons. However, to date no direct evidence has been provided that these proteins are complexed into an adheron-like particle. This report provides the first such evidence by showing that purification of hLAMP-1, under gentle conditions, results in the isolation of multiple bands of similar molecular weight within the fractions that contain anti-hLAMP-1 activity. PMID- 9215534 TI - Subcellular partitioning of transcription factors during osteoblast differentiation: developmental association of the AML/CBF alpha/PEBP2 alpha related transcription factor-NMP-2 with the nuclear matrix. AB - The subnuclear location of transcription factors may functionally contribute to the regulation of gene expression. Several classes of gene regulators associate with the nuclear matrix in a cell type, cell growth, or cell cycle related manner. To understand control of nuclear matrix-transcription factor interactions during tissue development, we systematically analyzed the subnuclear partitioning of a panel of transcription factors (including NMP-1/YY-1, NMP-2/AML, AP-1, and SP-1) during osteoblast differentiation using biochemical fractionation and gel shift analyses. We show that nuclear matrix association of the tissue-specific AML transcription factor NMP-2, but not the ubiquitous transcription factor YY1, is developmentally upregulated during osteoblast differentiation. Moreover, we show that there are multiple AML isoforms in mature osteoblasts, consistent with the multiplicity of AML factors that are derived from different genes and alternatively spliced cDNAs. These AML isoforms include proteins derived from the AML-3 gene and partition between distinct subcellular compartments. We conclude that the selective partitioning of the YY1 and AML transcription factors with the nuclear matrix involves a discriminatory mechanism that targets different classes and specific isoforms of gene regulatory factors to the nuclear matrix at distinct developmental stages. Our results are consistent with a role for the nuclear matrix in regulating the expression of bone-tissue specific genes during development of the mature osteocytic phenotype. PMID- 9215535 TI - A variation in the apolipoprotein C-III gene is associated with an increased number of circulating VLDL and IDL particles in familial combined hyperlipidemia. AB - Detailed plasma lipoprotein analyses were conducted on 16 familial combined hyperlipidemic (FCHL) probands, all their available family members (n = 106) together with 12 normolipidemic control families (n = 68), and the results were assessed in relation to a C1100-T polymorphism in exon 3 of the apoC-III gene. The frequency of the T1100 genotype (CT+TT) was significantly elevated in the probands relative to control subjects (0.64 vs. 0.36; P < 0.01) and was associated with elevated concentrations of plasma triglyceride (P < 0.02) and apoC-III (P < 0.03), VLDL cholesterol (P < 0.005), VLDL triglyceride (P < 0.009), IDL cholesterol (P < 0.01). and IDL triglyceride (P < 0.007). The T1100 genotype was also associated with elevations in VLDL-apoB (P < 0.005) and IDL-apoB (P < 0.04) indicating a relationship between this variation and an increased number of triglyceride-rich particles. These findings were confined to the hyperlipidemic members of the FCHL families and showed a strong genotype-status interaction (P < 0.001). It is considerable clinical relevance that the apoC-III gene may be acting as a modifier gene that is only expressed in the presence of other factors (e.g., increased VLDL flux, low LPL activity) and therefore may predispose those members of FCHL families carrying the T1100 allele to express the FCHL phenotype. PMID- 9215536 TI - Particle size determines the specificity of apolipoprotein E-containing triglyceride-rich emulsions for the LDL receptor versus hepatic remnant receptor in vivo. AB - Apolipoprotein E (apoE) is an important determinant for the uptake of triglyceride-rich emulsions and lipoproteins by the liver, and exerts affinity for both the LDL receptor (LDLr) and a distinct liver-specific recognition site. Our current aim was to assess the mechanism underlying the receptor-specificity of apoE-carrying lipoproteins. Triglyceride-rich emulsions were synthesized, with mean sizes of 50, 80, and 150 nm. These fractions efficiently acquired apoE from rat serum, and were processed by LPL in vivo with a similar efficiency. Upon injection of the [5H]cholesteryl oleate-labeled emulsions into rats, the liver association rate was positively correlated with particle size (24 +/- 2%, 54 +/- 1%, and 64 +/- 3% of the injected dose at 20 min after injection, respectively) and the liver uptake was predominantly exerted by parenchymal cells. The role of the LDLr in emulsion clearance was established in wild-type versus LDLr knockout mice. In the absence of the LDLr, an 8-fold increased serum half-life was observed for the small emulsion, concomitant with a 6- and 15-fold decreased uptake by the liver and adrenals at 60 min after injection, respectively. In contrast, the in vivo behavior of the large emulsion was independent of the LDLr. Both the ratio of apoE:C on the emulsions upon serum incubation and the alpha helical content of apoE were inversely correlated with particle size, indicating that these factors may be involved in the emulsion size-dependent receptor specificity in vivo. It is concluded that the contribution of the LDLr to the apoE-mediated clearance of emulsions by the liver and adrenals strongly increases with decreasing particle size, while large particles initially associate with a distinct liver-specific recognition site. As these emulsions mimic chylomicrons, we anticipate that the apoE-dependent metabolic behavior of chylomicrons (remnants) is largely dependent on their size. PMID- 9215537 TI - Biochemical and biophysical characterization of human recombinant lecithin: cholesterol acyltransferase. AB - We established a Chinese hamster ovary cell line that constitutively expresses up to 5 mg/L of human recombinant lecithin: cholesterol acyltransferase (rLCAT). We purified the rLCAT to > 96% purity, and characterized it along with plasma LCAT (pLCAT) biochemically and biophysically. The recombinant enzyme is more heavily glycosylated and more heterogeneous in its carbohydrate content than the plasma enzyme, as revealed by differences in molecular weight and pI isoforms, determined by mass spectrometry and isoelectric focusing. Recombinant LCAT is half as active enzymatically as pLCAT. The difference in activity is due to differences in the catalytic rates rather than in the apparent K(m) values, suggesting that the binding of the rLCAT to interfaces is not altered by its different glycosylation pattern. Despite these differences, rLCAT has essentially the same intrinsic tryptophan fluorescence emission spectrum and far-UV CD spectrum as pLCAT, indicating that the tertiary and secondary structures of both enzyme forms are very similar. Both enzyme forms have a propensity to self associate, and their multimers appear resistant to dissociation by SDS and dilution. The free energies of unfolding (delta G(H2O)) of rLCAT and pLCAT are 3.4 +/- 0.2 and 3.2 +/- 0.2 kcal/mol, respectively, as determined by guanidine hydrochloride denaturation monitored by fluorescence. These relatively low delta G(H2O) values support the notion that LCAT is capable of undergoing major conformational changes upon interaction with interfacial substrates. PMID- 9215538 TI - Influence of vesicle surface composition on the interfacial binding of lecithin:cholesterol acyltransferase and apolipoprotein A-I. AB - Interfacial binding affinities and capacities of lecithin:cholesterol acyltransferase (LCAT) and apolipoprotein A-I (apoA-I) for surfaces of different phosphatidylcholine (PC) composition, cholesterol content, and apolipoprotein content were measured with a vesicle model system. Native polyacrylamide gel electrophoresis was used to separate free protein from vesicle-bound protein. ApoA-I was isolated from human plasma and radiolabeled with iodine, whereas radiolabeled LCAT was purified from the media of Chinese hamster ovary cells that were transfected with human LCAT cDNA and incubated in the presence of [35S] cysteine and methionine. Bound and free radiolabeled LCAT and apoA-I were quantified by phosphorimage analysis. ApoA-I binding was not influenced by cholesterol content (14 mole%) but was influenced by the PC fatty acyl composition of the vesicle. PC species containing long chain, polyunsaturated fatty acids (PUFA) in the sn-2 position resulted in increased binding affinity (Kd = 75-177 nM) but reduced capacity (0.1-0.3 apoA-I/ 1000 PC) in comparison to sn-1 palmitoyl, sn-2 oleoyl PC (POPC, 750 nM and 1.4 apoA-I/1000 PC). LCAT binding affinity to POPC (2190 nM) was stronger in the presence of cholesterol (530 nM), and LCAT binding capacity was reduced (2.63 and 0.6 molecules LCAT/1000 PC, respectively). In comparison to POPC, LCAT binding affinity to sn-1 palmitoyl, sn-2 arachidonyl PC was stronger (611 nM) and binding capacity was reduced (0.7 LCAT/1000 PC). LCAT binding affinity and capacity to sn-1 palmitoyl, sn-2 eicosapentaneoyl PC (2041 nM, and 2.5 LCAT/1000 PC) were similar to those observed for POPC. We conclude that vesicle surface PC fatty acyl composition and cholesterol content significantly influence LCAT and apoA-I interfacial binding and therefore may alter LCAT enzymatic activity. PMID- 9215539 TI - Cloning and characterization of the rat apobec-1 gene: a comparative analysis of gene structure and promoter usage in rat and mouse. AB - ApolipoproteinB (apoB) mRNA editing involves a C to U deamination of the nuclear apoB mRNA and occurs in mammalian small intestine and in the liver of certain species. This reaction is mediated by a multicomponent enzyme complex that includes a catalytic subunit, apobec-1. Apobec-1 mRNA is widely expressed in the rat and mouse and is subject to tissue-specific regulation. In order to understand the basis for the species- and tissue-specific pattern of apobec-1 gene expression we have cloned and characterized the rat chromosomal apobec-1 gene. We demonstrate its structural organization and regulation in comparison to that of the mouse apobec-1 gene. The rat apobec-1 gene spans 16 kb and includes one untranslated (exon A) and five translated exons (exons 1-5). The mouse apobec 1 gene contains eight exons, of which the first three (exons A, B, C) are untranslated. Independent approaches demonstrated three distinct clusters of transcription initiation sites in both species, including exon A, the distal region of exon 1, and a separate group in the proximal region of exon 1. These transcription start sites generate three distinct mRNA species whose proportions differ in a tissue-specific fashion. Promoter-luciferase reporter constructions using regions flanking exon A and exon 1 of the rat apobec-1 gene identified two functional regions upstream of exon 1 that independently promote luciferase expression in transfected hepatoma and colon cancer cells. These data serve as a basis for an understanding of the regulation of apobec-1 gene expression, in particular the mechanisms that serve to restrict its expression to the gastrointestinal tract in higher mammals. PMID- 9215541 TI - Effect of lipid transfer activity and triglyceride hydrolysis on apolipoprotein B immunoreactivity in modified low density lipoproteins. AB - Consequences of alterations in the size and the lipid composition of low density lipoproteins (LDL) on apolipoprotein (apo) B immunoreactivity were analyzed using two distinct anti-apoB monoclonal antibodies (Mabs), i.e., 4G3, which recognizes an epitope closed to the binding site to the LDL receptor, and 2D8, which is directed against a distal region. Inhibition analysis revealed that the lipid transfer-mediated triglyceride enrichment of LDL isolated from 12 native human plasmas is associated with significant reductions in the expression of 2D8 and 4G3 epitopes (P < 0.05 in both cases). In contrast, triglyceride hydrolysis of triglyceride-rich LDL significantly increased apoB immunoreactivity as compared with non-lipolyzed counterparts (P < 0.05 with 2D8 and 4G3 Mabs). Among all the modified LDL fractions studied (n = 36), immunoreactivity of 2D8 and 4G3 epitopes correlated negatively and significantly with the triglyceride content (P < 0.01 in both cases), but with neither the size nor the other lipid parameters of LDL particles. Furthermore, changes in the triglyceride content of LDL correlated significantly with changes in apoB immunoreactivity after in vitro treatment with either lipid transfer activity alone (P < 0.01 with 2D8 and 4G3 Mabs) or lipid transfer activity combined with triglyceride hydrolysis (P < 0.01 with 2D8 and 4G3 Mabs). Finally, both the triglyceride content of native LDL and the total triglyceride level in 12 normolipidemic human plasmas correlated negatively and significantly with the expression of 2D8 epitope (P < 0.03 in both cases) and 4G3 epitope (P < 0.02 in both cases). It is concluded that triglycerides constitute a major determinant of the immunoreactivity of 2D8 and 4G3 apoB epitopes in LDL. PMID- 9215540 TI - Effects of legume consumption on serum cholesterol, biliary lipids, and sterol metabolism in humans. AB - Legume consumption appears to lower serum cholesterol and to increase cholesterol saturation of bile, but the mechanisms of these effects have not been established. We studied nine human subjects on a metabolic ward during two randomly ordered 6-7 week periods: one during consumption of a control diet and the other during consumption of the same diet with 120 gm mixed legumes substituted for foods having equivalent calories, fat, protein, and carbohydrate. Mean serum LDL cholesterol was significantly lower during legume consumption (126 vs. 138 mg/dl, P = 0.039). Legume consumption significantly increased mean cholesterol saturation index of gallbladder bile from 1.07 to 1.26 (P = 0.016), largely because of an increase in hepatic secretion of cholesterol from a mean of 90.2 mumol/h to 100.8 mumol/h (P = 0.042). Fecal neutral sterol output was unaffected by legumes, but fecal acidic sterols increased from a mean of 861 to 1202 mumol/day (P = 0.002) during legume consumption. Mean sterol balance became significantly more negative during legume consumption (-2140 vs. -2700 mumol/day, P = 0.037) indicating an increase in cholesterol synthesis. Mean fractional absorption of bile acid was lower during legume consumption than (0.947 vs. 0.960, P = 0.003). These data suggest that legume consumption lower LDL cholesterol by partially interrupting the enterohepatic circulation of bile acids and increases cholesterol saturation of bile by increasing hepatic secretion of cholesterol. PMID- 9215542 TI - Cloning, expression, and chromosomal localization of mouse liver bile acid CoA:amino acid N-acyltransferase. AB - A mouse liver lambda Zap XR cDNA library was screened using the coding region of human bile acid CoA:amino acid N-acyltransferase (BAT) cDNA as a probe. Ten positive clones were isolated and purified, two of which apparently possessed complete open reading frames for BAT based on sequence analysis of the ends of the cDNAs. One clone (mBAT#9) was selected for sequence analysis and characterization. mBAT#9 is 1869 basepairs in length and the full-length cDNA possesses a 189 basepair 5'-nontranslated region, an open-reading frame of 1260 basepairs, and a 404 basepair 3'-nontranslated region followed by a poly(A) tail. The open-reading frame codes for a 420 amino acid protein with a calculated molecular mass of 46,525 daltons. The structural gene for mBAT was mapped to mouse Chromosome 4. The amino acid sequence of mBAT is 69% identical and 84% similar to that of hBAT, and 86% identical and 95% similar to that of kan-1, a putative rat liver BAT. Enzymatically active mBAT was expressed in E. coli using the bacterial expression vector pKK233-2. Immunoblot analysis of expressed mBAT with rabbit anti-human BAT polyclonal antibodies detected a single protein with a molecular mass of approximately 45,000 daltons. Cytosol from cells transformed with mBAT#9/pKK233-2 possessed significant amounts of BAT-catalyzed conjugating activity with taurine as substrate but the expressed enzyme did not use glycine or fluoro-beta-alanine as substrates. The K(m) value for taurine was 1.9 mM +/- 0.1 mM in reactions with cholyl CoA as a cosubstrate. The specificity of mBAT for taurine as a substrate was confirmed by the demonstration, using HPLC electrospray ionization mass spectrometry, that mouse gallbladder bile contained only taurine conjugates of bile acids. The identification of the types of amino acid conjugates of bile acids present in mouse bile had not been previously reported. These results indicate that a taurine-specific form of BAT has been cloned and expressed from mouse liver. PMID- 9215543 TI - Asymmetric distribution of pause transfer sequences in apolipoprotein B-100. AB - Lipoprotein assembly requires a complex and regulated set of events that includes apolipoprotein B (apoB) translocation across the endoplasmic reticulum (ER) membrane, folding, and association with lipids. Unlike simple secretory proteins which are cotranslationally translocated directly into the ER lumen, nascent apoB contains pause transfer (PT) sequences that direct the transient stopping and subsequent restarting of its translocation, a phenomenon termed translocational pausing. During one particular translocational pause in apoB, the ribosome membrane junction and ER translocation channel have been shown to be altered in such a way as to expose the nascent polypeptide to the cytosol and direct a change in the proteins neighboring the nascent chain. In this study, we have experimentally identified the location and distribution of the translocational pauses that are present throughout apoB-100. We find that pause transfer sequences are distributed asymmetrically, clustering in three distinct domains: a) nine functional PT sequences appear in the amino terminal 20% of apoB, b) four more PT sequences occur just before the end of apoB-48, and c) an additional ten PT sequences are found between apoB-65-95. These clusters are interrupted by two lipid binding regions of approximately 100 kD each in which no PT sequences occur. The implications of this asymmetric distribution of PT sequences, and their correlation with previously hypothesized structural and functional domains of apoB, are discussed. PMID- 9215545 TI - A single copy of apolipoprotein B-48 is present on the human chylomicron remnant. AB - Individuals homozygous for the e2 allele encoding apolipoprotein E exhibit a remnant removal defect and accumulate substantial levels of intestinally derived particles containing apolipoprotein B-48 (apoB-48). Such lipoproteins were isolated from the plasma of E2/E2 individuals, and further purified by affinity chromatography using a polyclonal antibody specific for selective binding and removal of apoB-100-containing lipoproteins. The unbound lipoproteins, termed chylomicron remnants, were particles with average hydrated diameters of 31.2 nm as determined by dynamic light scattering. They contained apoB-48 and ApoE as their only protein components. The number of apoB-48 molecules on each lipoprotein was assessed by counting the number of antibody molecules bound to the surface of the chylomicron remnants, using either a monoclonal antibody specific for a single epitope on apoB-48 or a mixture of two such monoclonal antibodies specific for widely separated epitopes. The results of this analysis seem unambiguous: no more than one apoB-48 resides on the chylomicron remnant. Because apoB appears to be unable to transfer among lipoprotein particles, it may be inferred that nascent chylomicrons also contain a single copy of apoB-48. PMID- 9215544 TI - Activity of 3-ketosphinganine synthase during differentiation and aging of neuronal cells in culture. AB - Changes in the enzyme 3-ketosphinganine synthase activity in rat cerebellar granule cells in culture were studied during differentiation and aging. The enzyme activity with palmitoyl-CoA and stearoyl-CoA, precursors of, respectively, C18-sphinganine and C20-sphinganine, was studied on the total cell homogenate using radioactive serine. The enzyme assay was performed by thin-layer chromatography (TLC) separation of the enzyme reaction mixture, and the resultant radioactive 3-ketosphinganine was identified by chromatographic comparison with a chemically synthesized 3-ketosphinganine, and quantified by determination of the TLC radioactivity distribution on the basis of the radioactivity content of cell lipid extract that was determined by scintillation counting. Using palmitoyl-CoA, the enzyme activity progressively increased from 40 to 54 pmol of 3 ketosphinganine/mg cell DNA per min in the first 8 days and then progressively decreased, and was 39 pmol of C18-(3-ketosphinganine)/mg cell DNA per min at day 22 in culture. For stearoyl-CoA the enzyme activity was very low at day one and then increased to a constant value of about 15 pmol of C20-(3-ketosphinganine)/mg cell DNA per min. These results are in good agreement with the finding that the ganglioside species that contain C18-sphingosine increase during cell differentiation and remain constant during cell aging, while the ganglioside species that contain C20-sphingosine continuously increase during both cell differentiation and aging. PMID- 9215546 TI - Aqueous dissociation constants of bile pigments and sparingly soluble carboxylic acids by 13C NMR in aqueous dimethyl sulfoxide: effects of hydrogen bonding. AB - pKas for the acid dissociation of the carboxyl groups of bilirubin in water have been reported recently to be 8.1-8.4, or higher. These high values were attributed to intramolecular hydrogen bonding. They have led to suggestions that monoanions of bilirubin predominate at physiologic pH and are the species transported most readily into hepatocytes by carriers. Such high aqueous pKas are inconsistent with recent 13C nuclear magnetic resonance (NMR) measurements on mesobilirubin XIII alpha, done on aqueous solutions containing dimethyl sulfoxide. To investigate whether the presence of dimethyl sulfoxide leads to unreliable values when using 13C NMR spectroscopy to determine pKas of carboxylic acids that can undergo intramolecular hydrogen bonding, we measured the pKas of 13C-labeled fumaric, maleic, and phthalic acids in solutions containing up to 27 vol% dimethyl sulfoxide. In addition, we used 13C NMR to estimate the pKas of 2,2'-methylenebis[5-carbomethoxy-4-methylpyrrole-3-[1-13C] propanoic acid], a model for the two central rings of bilirubin. Our results show that 13C NMR of aqueous dimethyl sulfoxide solutions can be used with confidence to measure pKas of intramolecularly hydrogen-bonded carboxylic acids. They support our previous estimates for the pKas of bilirubin and confirm that intramolecular hydrogen bonding has little effect on the acidity of bilirubins in water. Together with previous studies and chemical arguments they strongly suggest that reported aqueous pKas of > 8, or even > 6, for the carboxyl groups of bilirubin are incorrect and that arguments used to rationalize them are questionable. PMID- 9215547 TI - Potent hypocholesterolemic activity of novel ureido phenoxyisobutyrates correlates with their intrinsic fibrate potency and not with their ACAT inhibitory activity. AB - The hypocholesterolemic activity for novel ureido fibrate analogues was found to be over 100-fold greater than for any "second-generation" fibrate in cholesterol fed rats. A comparison of 12 related analogues revealed that the optimal configuration for a urea-bridging region located between two aromatic rings consisted of a trisubstituted nitrogen, optimally substituted with a C7 alkyl chain and linked by dimethylene to a phenoxyisobutyrate moiety found in most fibrate analogues. The hypocholesterolemic potency of these compounds was found to correlate with their increased intrinsic fibrate activity as determined by the ability to induce omega-hydroxylase activity either in rat hepatocyte cultures or in vivo, and not with their 10-fold increased ACAT inhibitory potency when compared to other fibrates. The most active compound, 2-(4-(2-(N'-(2,4- difluorophenyl)-N-heptylureido)ethyl)phenoxy)-2-methylpropionic acid, referred to as (2), was found to induce omega-hydroxylase activity in hepatocytes at concentrations between 5 and 100 nM compared to 1-20 microM concentrations for bezafibrate, and lower serum VLDL+LDL cholesterol in rats at doses between 0.1 and 0.5 mg/kg per day compared to doses of 25-100 mg/kg per day for bezafibrate. Single-dose pharmacokinetic studies with 2 indicated that total drug exposure will be much lower at hypocholesterolemic doses due to the enhanced intrinsic activity, and may result in an improved safety profile for these novel trisubstituted ureido fibrate analogues in rats and humans compared to other fibrates. PMID- 9215548 TI - Quantitative and compositional changes in high density lipoprotein subclasses in patients with various genotypes of cholesteryl ester transfer protein deficiency. AB - High density lipoprotein (HDL) with and without apolipoprotein (apo) E was quantified and characterized in subjects with three genotypes of cholesteryl ester transfer protein (CETP) deficiency: the nonsense mutation in intron 14 (10 homozygotes and 5 heterozygotes); the missense mutation in the exon 15 (3 homozygotes and 9 heterozygotes); and the Int14A/D442G in 6 compound heterozygotes. ApoE-poor and apoE-rich HDL-cholesterol levels were elevated significantly in all genotypic groups with the decrease in CETP activity, indicating that both types of HDL-cholesterol can be a substrate for CETP. However, an unchanged or only slightly increased serum apoA-II level in each genotype indicated that the HDL particles with apoA-II are relatively resistant to CETP-mediated lipid transfer. Serum apoE-rich HDL level was considerably higher in the Int14A homozygotes than in the compound heterozygotes, in spite of similar apoE-poor HDL-cholesterol levels, which may indicate that apoE-rich HDL is a better substrate for CETP than apoE-poor HDL. Although the apoE-rich and apoE-poor HDL subclasses were similar in the accumulation of cholesteryl ester and depletion of triglyceride, the accumulation of free cholesterol was unique to apoE-rich HDL, indicating inhibited cholesterol esterification on this lipoprotein. Clinical laboratories should be aware of the discrepancy in HDL cholesterol measurements that comes from the different recoveries of apoE-rich HDL using commercial reagents. In conclusion, CETP deficiency causes considerable quantitative and compositional changes in HDL subclasses, reflecting a significant physiological role for CETP in HDL metabolism. PMID- 9215549 TI - Distribution of mixtures of bile salt taurine conjugates between lecithin cholesterol vesicles and aqueous media: an empirical model. AB - Bile salts are surfactants that partition into phospholipid bilayers. When liposomes or membranes are exposed to mixed solutions of bile salts, the more hydrophobic bile salt species associate preferentially with the lipid bilayer. As a consequence, in the aqueous phase, the free monomeric concentration of bile salt declines and the more hydrophilic species become relatively enriched. Above a critical saturating concentration of lecithin-associated bile salt, a phase transition occurs with loss of membrane integrity and formation of mixed micelles. In this paper we present a quantitative model which, for mixed solutions of bile salt taurine conjugates, predicts the distribution of bile salt monomers between large unilamellar vesicles composed of lecithin and cholesterol and the aqueous phase. The model is based on association isotherms for individual bile salts, determined by an ultrafiltration method with empirical curve fitting, and is critically dependent upon the observation that association coefficients of each bile salt are a function of the total bound bile salt/lecithin mole ratio. Given the concentrations of individual bile salts, lecithin and cholesterol, the model permits calculation of the membrane-bound bile salt/lecithin ratio and the concentration of each bile salt remaining free as soluble monomer in the aqueous phase, as well as the overall hydrophilic-hydrophobic balance (hydrophobicity index) of the bile salts remaining free in aqueous solution. Distribution data determined empirically for a variety of mixtures of bile salt taurine conjugates and large unilamellar vesicles of varying cholesterol:lecithin ratio agree closely with predictions. This model may be of value in predicting the physical, biological and toxic properties of mixed bile salt solutions. PMID- 9215550 TI - Structural requirements of Rhizobium chitolipooligosaccharides for uptake and bioactivity in legume roots as revealed by synthetic analogs and fluorescent probes. AB - Rhizobium chitolipooligosaccharides (CLOSs) are heterogeneous fatty acylated N acetyl glucosamine oligomers with variations in both the polar (hydrophilic) oligosaccharide head group and the non-polar (hydrophobic) fatty acyl chain. They trigger root hair deformation and cortical cell divisions in legume roots during development of the nitrogen-fixing root-nodule symbiosis. It has been proposed that only certain unique molecular species of CLOSs made by a particular rhizobia can elicit these responses on the corresponding legume host, suggesting that receptor-mediated perception of CLOSs serves as a basis of symbiotic specificity. We evaluated the relative symbiotic importance of the hydrophilic and hydrophobic structural domains of CLOSs by comparing the biological activities of CLOSs from wild type R. leguminosarum bv. trifolii ANU843 with that of various synthetic analogs. These tests were performed in axenic bioassays on the compatible symbiotic host, white clover (Trifolium repens) and the incompatible non-host legume, alfalfa (Medicago sativa). Fluorochrome-tagged derivatives of the native CLOSs and the analogs were also prepared in order to evaluate the uptake and localization patterns of these molecules within host root cells. The results indicate a direct link between uptake and biological activities of Rhizobium CLOSs on legume roots. The smallest CLOS analog taken up and biologically active on white clover and alfalfa was a N-fatty acylglucosamine, without an essential requirement of oligomerization, fatty N-acyl unsaturation, or acetate/sulfate functionalization. This suggests that N-fattyacylglucosamine is the common minimum structure required and sufficient for uptake and biological activity of CLOS glycolipids in these legumes, and that the various specific modifications of its polar head group and hydrophobic tail modulate its inherent ability to further express these activities, thus influencing which legumes are capable of responding to CLOSs rather than dictating their biological activities per se. PMID- 9215551 TI - A natural apolipoprotein A-I variant, apoA-I (L141R)Pisa, interferes with the formation of alpha-high density lipoproteins (HDL) but not with the formation of pre beta 1-HDL and influences efflux of cholesterol into plasma. AB - ApoA-I(L141R)Pisa is a naturally occurring apolipoprotein A-I variant that causes virtual absence of HDL in hemizygotes and hypoalphalipoproteinemia with half normal levels of HDL-cholesterol in heterozygotes. In this study we analyzed the distribution of HDL subclasses in plasmas of four hemizygotes for this mutation. We also investigated the abilities of these plasmas to esterify cholesterol and to promote cholesterol efflux. Residual apoA-I-containing lipoproteins in plasmas of hemizygotes for apoA-I(L141R)Pisa correspond to pre beta 1-LpA-I and small alpha-LpA-I. Unlike normal pre beta 1-LpA-I, pre beta 1-LpA-I of apoA I(L141R)Pisa hemizygotes was not converted into a larger alpha-migrating particle. Plasmas of apoA-I(L141R)Pisa hemizygotes were significantly reduced in their activity to esterify cholesterol in either endogenous or exogenous lipoproteins. Cholesterol efflux capacity was significantly lower than that of normal plasma. Efflux of [3H] cholesterol from radiolabeled fibroblasts into apoB depleted plasma of normal probands was biphasic with fast cholesterol efflux occurring in the first minute. Thereafter, cholesterol efflux was slow and unsaturable. After incubation with radiolabeled fibroblasts, efflux values of [3H]cholesterol into apoB-depleted plasma from normal controls and from apoA I(L141R)Pisa hemizygotes were indistinguishable at 1 min. Longer incubations with apoB-free plasma from apoA-I(L141R)Pisa hemizygotes did not, however, lead to the unsaturable increase in cholesterol efflux that was observed during incubations with apoB-free plasma of normolipidemic probands. Pre beta 1-LpA-I of apoA I(L141R)Pisa hemizygotes took up significantly less cell-derived [3H]cholesterol than pre beta 1-LpA-I of normal donors. We conclude that apoA-I(L141R)Pisa interferes with the formation of lipid-rich alpha-HDL but not with that of lipid poor pre beta 1-LpA-I. Very low concentrations of alpha-HDL in plasmas of apoA I(L141R)Pisa hemizygotes combined with reduced LCAT activity cause a decrease of the slow, unspecific, and LCAT-dependent components of cholesterol efflux into plasma. PMID- 9215552 TI - Sterol 27-hydroxylase: expression in human arterial endothelium. AB - Human endothelium obtained from both the aorta and the pulmonary artery has been evaluated for the presence of the messenger RNA coding for the expression of sterol 27-hydroxylase. Unique oligomers were designed to detect the mRNA by reverse transcription followed by the polymerase chain reaction. The amplified product was sequenced and was found to be identical to the published sequence for nucleotides 491 to 802 of the human sterol 27-hydroxylase cDNA. Northern blot analysis confirmed the presence of 27-hydroxylase mRNA in pulmonary artery and aortic endothelium. As part of these studies, enzymatic activity was assayed in cultured arterial endothelium using cholesterol as a substrate and isotope ratio gas-liquid chromatography-mass spectrometry to identify the metabolites, 27 hydroxycholesterol and 3 beta-hydroxy-5-cholestenoic acid, in the medium. Localization of sterol 27-hydroxylase to vascular endothelium indicates intracellular production of the biologically active metabolite 27 hydroxycholesterol. PMID- 9215553 TI - Characterization of a composite gradient gel for the electrophoretic separation of lipoproteins. AB - We describe a protocol for making a new type of gradient gel, the Composite gradient gel, that was designed to resolve plasma lipoproteins using nondenaturing gradient gel electrophoresis. The new gel format allows analysis both of high density lipoproteins (HDLs) and low density lipoproteins (LDLs) on the same gel. The gel gave highly repeatable (r2 = 0.999) size estimates. We compared lipoprotein phenotypes determined from the new gradient gel with those obtained using specialized HDL and LDL gradient gels. The comparisons indicated that the Composite gel gave lipoprotein particle size estimates for HDLs and LDLs that were virtually identical to those obtained, respectively, from the specialized HDL and LDL gradient gels. We measured median diameters, which reflect the distributions of absorbance, for LDLs and for HDLs and found that the Composite gel gave lipoprotein size distributions that were virtually identical to those measured using the specialized LDL and HDL gels. Finally, comparison of fractional absorbance for six lipoprotein size intervals obtained from the Composite and specialized gels revealed a close correlation (r2 = 0.828). Thus, it appears that both LDL and HDL size phenotypes may be evaluated simultaneously using a single gradient gel format. PMID- 9215554 TI - International Symposium on the Role of HDL in Disease Prevention: report on a meeting. PMID- 9215555 TI - Telomere maintenance without telomerase. AB - Telomeres are nucleoprotein structures at the ends of eukaryotic chromosomes that perform a number of vital functions. They allow a cell to distinguish between natural chromosome ends and chromosome breaks in order to delay the cell cycle and repair the broken end. Telomeres also compensate for the inability of DNA polymerase to replicate the chromosome completely. In most eukaryotes a special reverse transcriptase, telomerase, adds telomeric DNA repeats to the chromosome ends using an internal RNA template. However, evidence is accumulating for alternative elongation mechanisms in a variety of eukaryotes. In the yeast Saccharomyces cerevisiae, and possibly in humans, both of which normally use telomerase, a different mechanism can be used for chromosome length maintenance when telomerase is inactive or inactivated. Yeast apparently uses recombination for this purpose; the mechanism in humans is not known. Some insect and plant species, on the other hand, do not use telomerase as their primary mechanism for maintaining chromosome length. Drosophila makes use of specific retrotransposons for this purpose, while other dipterans use recombination. We summarize here the current knowledge of these alternative telomere elongation mechanisms. PMID- 9215556 TI - The conserved HR domain of the Drosophila suppressor 2 of zeste [Su(z)2] and murine bmi-1 proteins constitutes a locus-specific chromosome binding domain. AB - The related Drosophila Suppressor 2 of zeste [Su(z)2] and Posterior sex combs (Psc) proteins are both locus-specific chromosome binding proteins. They are found at many of the same polytene chromosome loci as other Polycomb-group proteins. The 1,365 amino acid Su(z)2 protein and the 1,603 amino acid Psc protein share a conserved 200 amino acid domain, the homology region (HR). To identify the protein domain responsible for locus-specific chromosome binding, we made a series of Hsp70:cDNA deletion constructs of the Sz(z)2 gene and transformed these into flies. We found that the HR is necessary and sufficient for Su(z)2 locus-specific polytene chromosome binding. The murine Bmi-1 protein also shares the conserved HR domain. When expressed in flies, the Bmi-1 protein showed a locus-specific chromosome binding pattern similar to that of the Su(z)2 and Psc proteins. These results argue that a locus-specific chromosome binding function resides in the HR domain. Other results show that a second, low affinity, non-specific chromosome binding function is localized outside the HR in the Su(z)2 protein, and that the Su(z)2 protein contains at least two nuclear localization signals. PMID- 9215557 TI - Subnuclear localization of S/MAR-binding proteins is differently affected by in vitro stabilization with heat or Cu2+. AB - The nuclear matrix, a proteinaceous entity thought to be a scaffolding structure that determines the higher order organization of eukaryotic chromatin, is usually prepared from intact nuclei by a series of extraction steps. In most cell types investigated, the nuclear matrix does not spontaneously resist these extractions, but must rather be stabilized before the application of extracting agents such as high salt solutions or lithium diiodosalicylate. We have examined the effect of two widely used stabilization procedures on the localization of nuclear matrix proteins. Four individual polypeptides were studied, all of which are scaffold or matrix-associated region (S/MAR)-binding proteins: SATB1, SAF-A/hnRNP-U, NuMA , and topoisomerase II alpha. Nuclei were isolated from K562 human erythroleukemia cells in a buffer containing spermine, spermidine, KCl and EDTA, and the nuclear matrix or scaffold was obtained by extraction with lithium diiodosalicylate after stabilization by heat treatment (37 degrees or 42 degrees C) or incubation with Cu2+ ions. When the localization of individual proteins was determined by immunofluorescent staining and confocal scanning laser microscopy, markedly different consequences of the two stabilization strategies became evident, ranging from a total maintenance of the localization (NuMA and topoisomerase II alpha) to a marked redistribution (SATB1 and SAF-A/hnRNP-U). Our results seem to indicate that a reevaluation of stabilization protocols employed for the preparation of the nuclear matrix is desirable, especially by performing morphological controls. PMID- 9215558 TI - Shared DNA sequences between the X and Y chromosomes in the tammar wallaby - evidence for independent additions to eutherian and marsupial sex chromosomes. AB - Marsupial sex chromosomes are smaller than their eutherian counterparts and are thought to reflect an ancestral mammalian X and Y. The gene content of this original X is represented largely by the long arm of the human X chromosome. Genes on the short arm of the human X are autosomal in marsupials and monotremes, and represent a recent addition to the eutherian X and Y. The marsupial X and Y apparently lack a pseudoautosomal region and show only end-to-end pairing at meiosis. However, the sex chromosomes of macropodid marsupials (kangaroos and wallabies) are larger than the sex chromosomes of other groups, and a nucleolus organizer is present on the X and occasionally the Y. Chromosome painting using DNA from sorted and microdissected wallaby X and Y chromosomes reveals homologous sequences on the tammar X and Y chromosomes, concentrated on the long arm of the Y chromosome and short arm of the X. Ribosomal DNA sequences were detected by fluorescence in situ hybridization on the wallaby Xp but not the Y. Since no chiasmata have been observed in marsupial sex chromosomes, it is unlikely that these shared sequences act as a pseudoautosomal region within which crossing over may occur, but they may be required for end-to-end associations. The shared region of wallaby X and Y chromosomes bears no homology with the recently added region of the eutherian sex chromosomes, so we conclude that independent additions occurred to both sex chromosomes in a eutherian and macropodid ancestor, as predicted by the addition-attrition hypothesis of sex chromosome evolution. PMID- 9215559 TI - A framework physical map of Drosophila virilis based on P1 clones: applications in genome evolution. AB - The analysis of patterns of genome evolution may help to evaluate the evolutionary forces that shape the composition and organization of the genome. Comparisons between the physical maps of divergent species can be used to identify conserved blocks of closely linked genes whose synteny is possibly under selective constraint. We have used in situ hybridization to determine the genomic position of 732 randomly selected clones from a bacteriophage P1 library of Drosophila virilis. The resulting map includes at least one clone in each of 69% of the subdivisions into which the D. virilis polytene chromosomes are divided. A subset of these clones was used to carry out a comparative physical analysis of chromosome 2 from D. virilis and from Drosophila montana. A number of discrepancies with the classical scenario of chromosome evolution were noted. The D. virilis P1 clones were also used to determine the physical relations between ten genes that are located in the X chromosome of Drosophila melanogaster between the markers crn (2F1) and omb (4C5-6). In this region, which is approximately 2 Mb in length, there have been at least six breakpoints since the divergence of the species, and six of the genes are found at widely scattered locations in the D. virilis X chromosome. However, a block of four functionally unrelated genes, including white, roughest, Notch, and dunce, seems to be conserved between the two species. PMID- 9215560 TI - Chromosomal homeologies between human, harbor seal (Phoca vitulina) and the putative ancestral carnivore karyotype revealed by Zoo-FISH. AB - We report on the construction of the first comparative Zoo-FISH map of a marine mammal. Zoo-FISH with DNA probes from a human chromosome-specific library to metaphase spreads of the harbor seal (Phoca vitulina) disclosed 31 conserved syntenic segments covering the complete autosomal complement and the X chromosome. Comparison with Zoo-FISH maps of other species reveals that the harbor seal shares a high degree of karyotypic homeology with the human complement and an even higher degree with the conordinal cat complement. These findings suggest that pinniped, felid and human karyotypes have maintained conserved complements. Based on data of Zoo-FISH and comparative cytogenetics, a Zoo-FISH map of the ancestral carnivore karyotype (Z-CAR) is proposed. Flow cytometry revealed that the DNA value of the harbor seal genome is 79% that of the human genome. PMID- 9215562 TI - Cytological study of the brown dominant position effect. AB - Classic recessive position effect variegation is related to inactivation of genes juxtaposed to heterochromatin and accompanied by cytologically visible heterochromatization (compaction) of the chromosome region containing these genes. Compaction and gene inactivation occur only in the rearranged homologue. In contrast to this, dominant variegation of the bw gene is known to involve transcriptional silencing in both the cis and trans copy, if they are paired. Our paper describes a cyto- logical approach to understanding this phenomenon. Analysis of salivary gland chromosomes carrying In(2R)bwVDe1 and In(2R)bwVDe2, evoking strong dominant bw variegation, has shown that in the rearranged homologues typical heterochromatization of the bw region and proximal neighbouring bands occurs. Heterochromatization was never observed on a normal homologue paired with a rearranged one. The insertion into the chromosome region 59E in the bwD strain is similar to pericentric heterochromatin. The insertion seems to induce heterochromatization of the neighbouring chromosome region and as a result the material of the insert and the 59E1-2 band join into a single block. When variegation is suppressed, the 59E1-2 band can be seen as a separate structure located proximal to the insert. This occurs in salivary gland polytene chromosomes of XYY males at 29 degrees C and in pseudonurse cell polytene chromosomes of otu11/otu11 females. All bands in the region of the non-rearranged homologue show normal morphology. Thus, although in all strains studied we observed heterochromatization in cis, the homologous regions in trans are not visibly affected. PMID- 9215561 TI - Fixation-dependent organization of core histones following DNA fluorescent in situ hybridization. AB - We have evaluated the effects of different DNA denaturation protocols commonly used in DNA fluorescent in situ hybridization (FISH) experiments on chromatin structure using indirect immunofluorescence. The use of antibodies to acetylated histones H3 and H4 demonstrates that the different procedures differ considerably in their extent of histone displacement. Procedures involving paraformaldehyde fixation were found to be compatible with the structural preservation of acetylated chromatin organization by indirect immunofluorescence. These results provide a basis for interpreting DNA FISH experiments aimed at determining chromatin organization of individual loci. PMID- 9215563 TI - Milestones in the molecular structure of the major histocompatibility complex. PMID- 9215564 TI - Cold adaptation parameters derived from cDNA sequencing and molecular modelling of elastase from Antarctic fish Notothenia neglecta. AB - The primary structure of an elastase from the Antarctic fish Notothenia neglecta (NE) was elucidated by molecular cloning and cDNA sequence analysis. The cDNA of interest was isolated from a cDNA library obtained from Notothenia's pyloric caeca. The amino acid sequence identity with mammalian elastases ranges between 53 and 64%, but interestingly reaches 79% with one isoform (CEB) of two recently isolated cod elastases. The most interesting changes distinguishing the model of NE, predicted from the three dimentional structure of the native porcine elastase (PE), concern the catalytic crevice located in the inter-domains region. These features might be involved in the adaptation to cold of the Antarctic elastase. PMID- 9215565 TI - Spectroscopic properties of an engineered maltose binding protein. AB - The maltose binding protein (MBP) has been site specifically labelled with a nitrobenzoxadiazole (NBD) group following mutation of a serine to a cysteine residue at position 337. The resulting protein shows a large ligand (maltose or beta-cyclodextrin) dependent increase in its steady-state fluorescence intensity. Analysis of the static (intensity and anisotropy) and dynamic (lifetime distributions) fluorescence of the NBD label as well as the tryptophan residues in both ligand-bound and ligand-free states of this molecule reveals complex multi-component decays that are interpreted in terms of a ligand-induced solvent shielding mechanism. In the context of the known crystal structures of the various forms of the maltose-binding protein (MBP), ligand-dependent changes in both the fluorescence parameters as well as the circular dichroism spectra of the NBD group are interpreted by a twisted intramolecular charge-transfer (TICT) mechanism, wherein ligand binding locks the NBD group into a conformation that prevents efficient relaxation of the excited state. PMID- 9215566 TI - A consensus residue analysis of loop and helix-capping residues in four-alpha helical-bundle proteins. AB - A statistical study was performed on a set of proteins which adopt the four-alpha helical-bundle tertiary motif in order to determine amino acid occurrences at helix-capping and loop positions. Eight X-ray crystal structures from the Brookhaven Protein Data Bank (PDB) were examined and N", N', Ncap, Ccap, C' and C" residues were assigned. In addition, a set of 55 protein sequences for the analogous proteins from different strains and species was taken from the Protein Information Resource and Swiss-Prot databanks. The residues at the capping and loop positions in this expanded data set were deduced by aligning these sequences with those from the PDB files. Similar trends were observed in the two data sets. In general, polar residues were predominant in the loops, although aromatic residues were also fairly common. Glycine, a highly flexible residue with an excellent 'helix-breaking' ability, was very common at the Ccap, C' and C" residues. Proline, which can force sharp turns in the direction of a peptide backbone, was only common at the N" residue. Residues which can participate in the N-capping box motif were found with high frequency. Capping motifs at the helix C-termini (Schellman and alphaL motifs) were also somewhat common, while another helix N-terminal stabilizing motif, the hydrophobic stable, was not common. The data presented in this study should prove useful for applying the 'consensus residue' approach to the de novo design of loop regions in helical bundle proteins. PMID- 9215567 TI - Identification of membrane spanning beta strands in bacterial porins. AB - The membrane assembly of outer membrane proteins is more complex than that of transmembrane helical proteins owing to the intervention of many charged and polar residues in the membrane. Accordingly, the predictive accuracy of transmembrane beta strands is considerably lower than that of transmembrane alpha helices. In this paper we develop a set of conformational parameters for membrane spanning beta strands. We formulate an algorithm to predict the transmembrane beta strands in the family of bacterial porins based on the conformational parameters and surrounding hydrophobicities of amino acid residues. A Fortran program has been developed which takes the amino acid sequence as the input file and gives the predicted transmembrane beta strand as output. The present method predicts at an accuracy level of 82% for all the bacterial porins considered. PMID- 9215568 TI - A model of the lutropin/choriogonadotropin receptor: insights into the structural and functional effects of constitutively activating mutations. AB - A model of the seven transmembrane helical domain (7-TM) of the human lutropin receptor was constructed from the 2D electron density map of bovine rhodopsin and a set of geometric parameters derived from a published analysis of 204 G-protein coupled receptor sequences. The Self-Consistent Ensemble Optimization method was applied to predict overall side chain packing. The model is consistent with the general helical packing properties expected of transmembrane proteins and suggests possible structural and functional effects of constitutively activating mutations. A tightly packed hydrophobic cluster formed between the intracellular halves of TM5 and TM6, as well as a specific H-bonding network formed between the central regions of TM6 and TM7, is proposed to be critical for stabilizing the inactive form of the receptor. The model suggests that single activating mutations perturb the specific interactions of TM6 with TM5 and TM7, either by disrupting the hydrophobic packing between TM5 and TM6, or by weakening the H bonds between TM6 and TM7. PMID- 9215569 TI - Molecular modelling of the nicotinic acetylcholine receptor transmembrane region in the open state. AB - A model of the nicotinic acetylcholine receptor transmembrane region has been constructed which may represent the channel in its open-state. The positions of helices flanking the ion channel match those observed by electron microscopy and previously reported by others. Residues labelled, mutated or by other means known to have a strong influence on ion flux are each accessible from the lumen of the modelled channel. The model provides new insights into our current understanding of the ion channel structure, and suggests some novel explanations for the results of labelling and mutation studies such as those involving ion channel blockers and residue-dependent changes in ion selectivity. PMID- 9215570 TI - The 'Asx-Pro turn' as a local structural motif stabilized by alternative patterns of hydrogen bonds and a consensus-derived model of the sequence Asn-Pro-Asn. AB - Analyses of databases derived from the Brookhaven Protein Data Bank have identified a set of related turn structures formed by the sequence Asx-Pro Xxx(n). In a variety of flanking structural contexts, more than 60% of Asx-Pro sequences adopt a turn conformation stabilized by a set of alternative hydrogen bonds among the side chain O delta and backbone C = O carbonyl oxygens of Asx (residue i) and the backbone NH of residues i + 2, i + 3 and in some cases i + 4. In contrast, the structures adopted by Ser-Pro, His-Pro and other Xxx-Pro sequences reflect more heterogeneous hydrogen-bonding patterns. As expected, structures formed by Asx-Pro-Asx are similar to those formed by Asx-Pro-Xxx(n), but in some cases additional hydrogen bonds are formed between the Asx side chains. Hydrogen bond patterns within Asx-Pro and Asn-Pro-Asn turns are consistent with published NMR studies of helical (Asn-Pro-Asn-Ala)n peptides, indicating that a consensus structure reflecting these hydrogen bonds can serve as a partial model of the Asn-Pro-Asn-Ala tetrapeptide repeats of Plasmodium falciparum circumsporozoite protein. PMID- 9215572 TI - Hyperthermostable mutants of Bacillus licheniformis alpha-amylase: thermodynamic studies and structural interpretation. AB - This paper provides further understanding of the thermodynamic and structural features determining the stability of Bacillus licheniformis alpha-amylase (BLA) at two crucial positions, His133 and Ala209. Results of protein modelling and saturated site-directed mutagenesis at position 133 and 209 have been reported in a previous paper (Declerck et al., 1995, Prot. Engng, 8, 1029-1037). In the first part of the present work, evidence is presented supporting the hypothesis that the stabilizing mutations reduce the rate of initial unfolding of the enzyme during the reversible step of the inactivation reaction and do not modify the irreversible processes undergone subsequently by the unfolded molecules. In the second part, we have examined the three-dimensional structure of BLA which has been determined recently by X-ray analysis (Machius et al., 1995, J. Mol. Biol., 246, 545-559). This analysis showed that our previous predictions made from molecular modelling were partly correct. At position 209, the effect of the stabilizing substitutions can be explained by a groove-filling effect reinforcing the hydrophobic packing between two helices of the central domain, while preserving a well-ordered water structure at the surface. At position 133, the stabilizing substitutions must compensate the loss of the hydrogen bond network in which the original histidine side-chain is involved; this compensation could be achieved through enhanced hydrophobic side-chain interactions within the beta sheet where residue 133 is located, which correlates with the propensity of the residue to form and maintain a beta-strand conformation of the main chain at this position. PMID- 9215571 TI - Recognition of phage-expressed peptides containing Asx-Pro sequences by monoclonal antibodies produced against Plasmodium falciparum circumsporozoite protein. AB - The immunodominant region of the Plasmodium falciparum circumsporozoite protein is comprised mainly of a series of tetrapeptide repeats that can, depending on the starting cadence chosen, be described as (NANP)n, (ANPN)n, (NPNA)n or (PNAN)n in one-letter amino acid code. Data from several studies suggest that the NPNA cadence alone is structurally correct, in that each NPNA tetrapeptide effectively forms a structural unit initiated by an Asx-Pro turn. To explore this idea further and to assess the immunological relevance of peptide conformation as it relates to the cadence of these tetrapeptide repeats, we used ELISA to compare the abilities of monoclonal antibodies (MAbs) produced against P. falciparum sporozoites to recognize repeat-related heptapeptides expressed on the surface of filamentous bacteriophage. Having included representatives of both NANP and NPNA cadences and other peptides in which the number and location of Asx-Pro sequences varied, we provide evidence that Asx-Pro sequences play an important role in peptide conformation and antibody recognition, that peptide conformation is influenced by the cadence of the tetrapeptide repeats and that peptide conformation is important to the abilities of these MAbs to recognize their epitopes. PMID- 9215573 TI - Regulation of trypsin activity by Cu2+ chelation of the substrate binding site. AB - Lysine 188 of trypsin was replaced with histidine in order to create a metal chelation site in the substrate binding pocket of this protease, built in a metal binding 'switch,' and to be able to modulate its activity at lower pH. The catalytic properties of wild-type and mutant trypsin were measured with tetrapeptide substrates containing a nitroanilide leaving group and whole native protein substrate: beta-casein. The results obtained reveal that K188H mutation does not affect catalytic efficiency, raising only slightly (from 6 to 8) the arginine/lysine preference of the mutant and increasing 1.8- and 1.2-fold the second-order rate constant k(cat)/Km for arginine- and lysine-containing substrates, respectively. Compared with wild-type trypsin, K188H mutant shows, in the absence of Cu2+, a different catalytic activity pattern as a function of pH. The addition of Cu2+ to trypsin K188H induces a 30-100-fold increase in Km, while k(cat) is scarcely decreased. The hydrolytic activity of this mutant can be fully restored by addition of EDTA. In contrast to a chelating active site, a novel mode of metal-dependent inhibition activity of trypsin with copper is presented. As suggested by molecular modelling studies, the substrate binding pocket of the protease is considerably perturbed by vicinal chelation. More generally, this type of transition metal chelate may present wider possibilities of trypsin activity and specificity modulation than the previously accomplished chelation of a histidine moiety from a catalytic triad. PMID- 9215574 TI - Characterization of the malonyl-/acetyltransacylase domain of the multifunctional animal fatty acid synthase by expression in Escherichia coli and refolding in vitro. AB - cDNAs of various lengths encoding the second domain of the multifunctional fatty acid synthase (FAS) have been expressed in Escherichia coli and the recombinant proteins refolded in vitro to catalytically active monomeric malonyl /acetyltransacylases. FAS residues 428-487, previously thought to represent the amino terminus of the malonyl-/acetyltransacylase, can be omitted from the recombinant enzyme with no loss in catalytic activity. This shortened transacylase, consisting of FAS residues 488-809, can be repeatedly denatured and renatured in vitro with reproducibly high recovery and no loss in specific activity. When expressed as a soluble enzyme in Spodoptera frugiperda cells, this transacylase has the same specific activity as the enzyme that has been refolded in vitro. The refolded transacylase consisting of FAS residues 488-809, but not the longer enzyme consisting of residues 428-815, can be crystallized readily. These results suggest that FAS residues 428-487, previously thought to represent the amino terminus of the malonyl-/acetyltransacylase, are not required for catalysis of the transacylase reaction. This region of the FAS is less well conserved than the transacylase catalytic domain and may constitute an extended structural linker that facilitates the functional interaction between the transacylase and acyl carrier protein domains. PMID- 9215575 TI - Construction of a homodimeric dihydrofolate reductase-thymidylate synthase bifunctional enzyme. AB - A gene encoding a bifunctional homodimeric dihydrofolate reductase-thymidylate synthase (DHFR-TS) was constructed by destroying the stop codon of Escherichia coli dihydrofolate reductase (DHFR) and joining the coding sequences of the monofunctional enzymes by a five amino acid linker. The protein was designed to mimic features of active site proximity and electrostatics in the protozoan DHFR TSs which are believed to be important in channeling of the DHFR substrate, H2folate, to TS. The genetically engineered catalytically active homodimeric bifunctional DHFR-TS was expressed, purified and characterized. The component activities of the purified bifunctional enzyme had kinetic properties similar to those of the monofunctional TS and DHFR, but unlike the authentic bifunctional enzymes from protozoa this enzyme did not kinetically channel dihydrofolate from DHFR to TS. PMID- 9215576 TI - Importance of a conserved phenylalanine-35 of cytochrome b5 to the protein's stability and redox potential. AB - Phenylalanine-35, which is a residue of the hydrophobic patch on the surface of cytochrome b5, has been mutated into Tyr35, His35 and Leu35 to elucidate the functions of the Phe35 and give further insight into the roles of the hydrophobic patch and/or aromatic network. The effects of these mutations on the heme environment, denaturation towards heating and the denaturant urea, redox potential and stability of protein were studied. The relative stability of cytochrome b5 and its mutants towards heating has the order Phe35Tyr > wild type > Phe35Leu > Phe35His in the oxidized state and wild type > Phe35Tyr > Phe35Leu > Phe35His in the reduced state. All the mutants exhibit decreased reduction potentials: Phe35Tyr -66 mV, Phe35His -51 mV and Phe35Leu -28 mV, which are more negative than that of the wild type. The order of redox potential reflects the relative stability in the oxidized and reduced states. A method of producing multiple mutants at a single site of a gene is also described for the first time. PMID- 9215577 TI - Trp128Tyr mutation in the N-lobe of recombinant human serum transferrin: 1H- and 15N-NMR and metal binding studies. AB - The conserved Trp residue within helix 5 of the N-lobe of human serum transferrin (hTF/2N, 40 kDa) has been mutated to Tyr. NMR and CD spectra and energy calculations show that the mutation causes little perturbation of the overall structure of hTF/2N although the chelating agent Tiron removed Fe3+ from the mutant protein about three times faster than from wild-type hTF/2N. 1H-NMR resonances of residues in the Leu122-Trp128-Ile132 hydrophobic patch are assigned both by ring current calculations and with the aid of the mutation. [1H, 15N]-NMR resonances for 11 of the 14 Tyr residues were observed in the spectra of 15N-Tyr hTF/2N and a resonance for Tyr128 was assignable in spectra of the mutant. The 15N resonance of Y128 was sensitive to oxalate and Ga3+ binding, and Ga3+ binding perturbed 15N resonances for most of the Tyr residues. Since these are well distributed over the N-lobe, it can be concluded that metal-induced structural changes are not merely local to the binding site. PMID- 9215578 TI - Recombinant human TIMP-3 from Escherichia coli: synthesis, refolding, physico chemical and functional insights. AB - Matrix metalloproteinases are inhibited by a growing family of specific tissue inhibitors, TIMPs. The cDNA of the third member of the family, TIMP-3, was obtained by using a reverse transcription-polymerase chain reaction (RT-PCR) to amplify the corresponding mRNA from human placenta. Cloning and expression of the TIMP-3 were performed in Escherichia coli as a fusion protein with a 36 amino acid N-tail containing a His cluster. In the host vector system, rhTIMP-3 was stored intracellularly in its denatured, insoluble form in inclusion bodies. Slow dilution of denaturing and reducing agents, from rhTIMP-3 His bound to a metal affinity solid phase, was followed by partial acid removal of the N-tail, which leaves a residue of four amino acids. Circular dichroism, fluorescence and second derivative UV spectroscopic analyses supported correct refolding of the recombinant and zymography showed inhibition of both MMP-2 and MMP-9 gelatinolytic activities. The role of the C-terminus, which has closer homology with TIMP-2 than TIMP-1, was also investigated: a C-truncated mutant, similarly cloned and expressed in E. coli, shows complete lack of inhibitory activity on MMP-9, still retaining some on MMP-2. The described protein engineering shows high yield of active inhibitor, unglycosylated as in the native form. PMID- 9215579 TI - Heterogeneity in recombinant HIV-1 integrase corrected by site-directed mutagenesis: the identification and elimination of a protease cleavage site. AB - Purified recombinant human immunodeficiency virus type 1 (HIV-1) integrase and certain deletion mutants exhibit heterogeneity consistent with proteolysis at a site close to the C-terminus. Electrospray ionization mass spectrometric analysis indicated that proteolytic cleavage generated a protein missing five residues from the C-terminus. PCR mutagenesis of amino acids on either side of the cleavage site identified two changes which were subsequently shown to prevent clipping when proteins were expressed and purified from Escherichia coli: the substitution of Arg284, the residue on the C-terminal side of the cleavage site, by either glycine or lysine. The introduction of either of these mutations into full-length integrase did not affect in vitro 3' processing or strand transfer activities. Thus, the incorporation of either of these mutations is likely to be beneficial when homogeneity of HIV-1 integrase is a concern, as in crystallographic or nuclear magnetic resonance spectroscopic experiments. PMID- 9215580 TI - Affinity selection of a camelized V(H) domain antibody inhibitor of hepatitis C virus NS3 protease. AB - The HCV genome encodes, within the NS3 gene, a serine protease whose activity specifically cleaves the viral polyprotein precursor. Proteolytic processing of HCV polyprotein precursor by the viral NS3 proteinase is essential for virion maturation and designing specific inhibitors of this protease as possible anti viral agents is a desirable and practical objective. With a view to studying both the function of HCV NS3 protease and to designing inhibitors of this enzyme, we directed our interest towards engineering macromolecular inhibitors of the viral protease catalytic activity. We describe here the affinity-selection and biochemical characterization of one inhibitor, cV(H)E2, a 'camelized' variable domain antibody fragment, isolated from a phage displayed synthetic repertoire, which is a potent and selective inhibitor of proteolysis by the NS3 enzyme. In addition to being useful as a biological probe to study the function of HCV protease, this inhibitor can serve as a potential pharmacophore model to design antivirals. Moreover, the results suggest a way of engineering improved human derived small recognition units tailored for enzyme inhibition. PMID- 9215581 TI - Occurrence of S and F1C/S-related fimbrial determinants and their expression in Escherichia coli strains isolated from extraintestinal infections. AB - The presence of S and F1C/S-related fimbrial determinants was determined in 462 E. coli strains obtained from different extraintestinal infections and in 162 control isolates of E. coli by using two different DNA probes: an oligonucleotide probe consisting of three oligonucleotides that bind specifically to the S adhesin gene and a polynucleotide probe which is not able to distinguish between S, F1C, and S-related sequences. The expression of S and F1C phenotypes was tested by dot enzyme immunoassay with the corresponding monoclonal antibodies. S fimbriae genotypes were observed more frequently in septic (25%) and urinary (12%) isolates of E. coli than in faecal and water isolates (1%) and often occurred together with O2, O6, O18 and O83 antigens. F1C/S-related fimbrial DNA was detected with a higher frequency in UTI isolates (26%) than in septic (16%) and faecal (10%) isolates and was most frequently associated with O4, O6, and O75 serotypes. Since the production of S and F1C fimbriae was comparatively rare in all clinical and control isolates of E. coli, DNA hybridization assays which allow the sensitive and specific detection of fimbrial determinants even in the absence of their expression are preferable to phenotypic assays. PMID- 9215582 TI - Molecular analysis of naturally occuring ermC-encoding plasmids in staphylococci isolated from animals with and without previous contact with macrolide/lincosamide antibiotics. AB - A total of 16 epidemiologically unrelated macrolide-resistant staphylococcal isolates of various animal origins were investigated for the molecular basis of macrolide resistance with respect to previous contact of their host animals with macrolides and lincosamides. All isolates carried ermC-encoding plasmids of 2.3 4.0 kbp. The eight plasmids of staphylococci from animals which had not received macrolides or lincosamides showed inducible ermC gene expression and did not exhibit alterations in the ermC regulatory region. The remaining eight plasmids expressed the ermC gene constitutively. Six of these plasmids were from staphylococci from animals which had received tylosin or spiramycin as feed additives or lincomycin for therapeutic purposes. All constitutively expressed ermC genes revealed either sequence deletions or sequence duplications in their ermC regulatory region, as detected by a PCR assay and by sequence analysis. These sequence deletions and duplications found in naturally occurring plasmids corresponded closely to the mutations seen in the ermC-encoding plasmids after growth of an inducibly resistant strain in the presence of non-inducing macrolides or lincosamides under in vitro conditions. PMID- 9215583 TI - Hyporesponsiveness of inflamed human gingival fibroblasts from patients with chronic periodontal diseases against cell surface components of Porphyromonas gingivalis. AB - Inflamed human gingival fibroblasts (HGF) of patients with chronic periodontal diseases have less active interleukin-8 (IL-8) production compared with normal HGF of volunteers with healthy gingival tissues, after stimulation with Porphyromonas gingivalis surface components such as fimbriae, lipopolysaccharide (LPS) and its lipid A, but not LPS or lipid A from other bacterial species. A decrease in number of specific binding sites for P. gingivalis fimbrial molecules in inflamed HGF is also observed by Scatchard plot analysis. A short exposure (6 h) to P. gingivalis LPS resulted in significant potentiation of the LPS-dependent IL-8 production in normal HGF, whereas a long exposure (48 h) to the LPS significantly reduced IL-8 production. Tyrosine phosphorylation of proteins of 127 kDa and 186 kDa in inflamed HGF stimulated with P. gingivalis fimbriae or its LPS was observed by immunoblotting, and these two phosphoproteins were termed tolerance-induced protein, TIP. Protein bands of 45 kDa which bound to radioiodinated P. gingivalis fimbriae in the presence and absence of fetal bovine serum (FBS), and major 73-kDa and minor 30-kDa and 45-kDa bands which bound to radioiodinated P. gingivalis LPS in the presence of FBS in normal and inflamed HGF were observed by using photocrosslinking. These findings suggest that the hyporesponsiveness of HGF induced by a prolonged exposure to P. gingivalis may emerge because of HGF damage or result from host defense in chronic periodontal lesions. PMID- 9215584 TI - A simple Trypanosoma cruzi enzyme-linked immunoassay for control of human infection in nonendemic areas. AB - An enzyme linked immunosorbent assay (ELISA) was developed for detecting IgM and IgG antibodies against Trypanosoma cruzi in blood bank donors from endemic or nonendemic areas. A crude extract of trypomastigotes from cultures was used as antigen. A total of 494 serum samples from patients with acute, congenital, or chronic form of Chagas' disease, and from healthy French individuals were studied. The sensitivity of the ELISA was determined with 89 serum samples from chagasic patients and was evaluated to 98.8%. The specificity was determined with 405 serum samples from French blood transfusion centers donors and evaluated to 98.3%. Two hundred and eighty-five serum samples from blood donors from Argentina and Brazil were also tested. Furthermore, in order to assess the absence of cross reactivity with other protozoan infections, we studied 86 serum samples including (i) 32 individuals with cutaneous leishmaniasis living in a T. cruzi endemic region of Bolivia, and (ii) 54 patients from nonendemic area for Chagas' disease, 19 of them with kala-azar and 35 others with malaria. PMID- 9215585 TI - Ehrlichia sennetsu groE operon and antigenic properties of the GroEL homolog. AB - A clone expressing an immunoreactive 55-kilodalton (kDa) protein of Ehrlichia sennetsu, the causative agent of human Sennetsu ehrlichiosis, was isolated from a gene library of this organism. Sequence analysis of the DNA insert revealed two open reading frames, encoding proteins of 10,620 and 58,225 kDa, respectively. These deduced amino acid sequences were homologous to those of the GroES and GroEL heat shock proteins (HSP) of other bacteria, respectively. Phylogenetic trees based on GroES and GroEL homologs of several bacteria including E. sennetsu showed a relationship similar to that based on 16S rRNA gene sequences. The recombinant and native 55-kDa proteins of E. sennetsu, GroEL homolog, reacted with a monoclonal antibody (SPA807) which recognizes a homologous sequence between human and mycobacterial HSP60 and a polyclonal antibody (SPA804) to cyanobacteria HSP60, but not with antibodies to HSP60 of several other organisms used. Furthermore, anti-recombinant E. sennetsu 55-kDa protein antibody prepared in a rabbit was reactive to HSP60 antigens of other Ehrlichia and Rickettsia species, but not GroEL of E. coli. The recombinant 55-kDa protein would be a useful tool for studying the role of this antigen in the immune response to E. sennetsu infection. PMID- 9215586 TI - Characterization of Bacillus thuringiensis isolated from infections in burn wounds. AB - Four strains of Bacillus thuringiensis were isolated from infections in burn wounds and from water used in the treatment of burn wounds. The strains produced large parasporal inclusion bodies composed of 141, 83, and 81 kDa protoxins. The four strains were tested for insecticidal activity against larvae of Pieris brassicae and Aedes aegypti but showed no activity; Vero cell assays for the production of enterotoxins were also negative. Attempts to classify the strains according to flagellar H-serotype showed them all to be non-flagellated. Apart from two occupational health accidents that occurred during the handling of highly concentrated B. thuringiensis fluids, this is the first report of B. thuringiensis causing non-gastrointestinal clinical infection in immunosuppressed patients. PMID- 9215587 TI - Shiga toxin induces medullary tubular injury in isolated perfused rat kidneys. AB - To investigate the potential direct nephrotoxicity of Shiga toxin, a putative mediator for hemolytic uremic syndrome, purified toxin (10(-11) M) was added to isolated rat kidneys perfused for 160 min with a Krebs-Henseleit acellular medium enriched with albumin and amino acids. Kidney function and morphology were examined after perfusion with the Shiga toxin vs controls. Shiga toxin did not significantly alter renal perfusion flow, glomerular filtration rate, or tubular sodium reabsorption, but it significantly increased urinary protein excretion (from 61 +/- 23 to 169 +/- 28 microg/min, P < 0.01). On renal morphologic study, Shiga toxin did not induce gross glomerular damage but increased markedly the injury to the medullary thick ascending limbs. In conclusion, Shiga toxin is toxic to rat kidneys ex vivo and in the absence of platelets. Renal damage is manifested by proteinuria and medullary tubular injury. The distribution of this injury suggests a possible synergism between local medullary hypoxia and the toxic tubular or endothelial effects of the toxin. These effects may play a pathogenic role in the tubulo-interstitial injury observed in hemolytic uremic syndrome associated with severe renal failure. PMID- 9215588 TI - Genotypic differentiation of Gardnerella vaginalis by amplified ribosomal DNA restriction analysis (ARDRA). AB - In total 34 strains of Gardnerella vaginalis were analyzed with various molecular techniques in order to find the possibility of dividing this single species into different genotypes. Classical ribotyping, PCR-ribotyping and restriction analysis of 16S-23S rRNA intergenic spacer sequences were all unsuccessful in genotype differentiation of these bacteria. Only amplified ribosomal DNA restriction analysis (ARDRA) was suitable in recognizing different G. vaginalis genotypes. At least 3-4 genotypes were identified with different restriction enzymes, some of which showed a prevalent distribution in certain of the centers from which they were collected. Although in this study no correlation was found between bacterial vaginosis and any of the genotypes identified, the ARDRA method could prove to be a useful tool for studying the etiopathology and epidemiology of G. vaginalis. PMID- 9215589 TI - Expression of intercellular adhesion molecule-1 (ICAM-1) on vascular endothelial cells and renal tubular cells in the generalized Shwartzman reaction as an experimental disseminated intravascular coagulation model. AB - The participation of adhesion molecules in systemic vascular injuries of the generalized Shwartzman reaction was studied. The generalized Shwartzman reaction was induced in mice by two consecutive injections of lipopolysaccharide. Intercellular adhesion molecule-1 (ICAM-1) was expressed on vascular endothelial cells, renal tubular cells and alveolar wall in generalized Shwartzman reaction induced mice. The preparative injection of lipopolysaccharides induced ICAM-1 expression in those cells, and the provocative injection of lipopolysaccharides for the generalized Shwartzman reaction augmented it further. The simultaneous administration of anti-gamma interferon antibody with the preparative injection of lipopolysaccharides completely inhibited ICAM-1 expression on vascular endothelial cells. The injection of recombinant gamma interferon in replacement of lipopolysaccharides resulted in ICAM-1 expression. The administration of anti ICAM-1 antibody together with the provocative injection of lipopolysaccharides significantly blocked the apoptosis of vascular endothelial cells in generalized Shwartzman reaction-induced mice. It was suggested that ICAM-1 expression on vascular endothelial cells might be involved in systemic vascular injuries of the generalized Shwartzman reaction, and that it might be regulated by gamma interferon. PMID- 9215590 TI - Immunization with recombinant Trypanosoma cruzi ribosomal P2beta protein induces changes in the electrocardiogram of immunized mice. AB - Molecular expression cloning techniques revealed that patients with severe chronic Chagas heart disease showed a strong humoral response against the cloned C-terminal portion of the Trypanosoma cruzi ribosomal P2beta protein, previously named JL5. The main linear epitope of this polypeptide was mapped to the 13 C terminal amino acid sequence EEEDDDMGFGLFD (named R13), which is almost identical to the mammalian ribosomal P consensus sequence EESDDDMGFGLFD (named H13). Enzyme linked immunosorbent assay measurements demonstrated that sera from patients with chronic Chagas heart disease presented a very specific anti-P humoral response with high anti-R13, but low H13 antibody levels. We attempted to develop an animal model that would reproduce, at least partially, two features of the human infection: (1) the serological pattern of the anti-P response, and (2) specific cardiac symptoms. To this effect, mice were immunized with T. cruzi P2beta recombinant protein. Immunization reproduced the typical anti-P antibody profile defined for chronic infections, but did not induce cardiac inflammatory lesions. However, it altered significantly the electrocardiograms of immunized mice. It is suggested that this assay represents a functional test for assessing the biological activity of antibodies against T. cruzi ribosomal P protein on cardiac muscle. PMID- 9215591 TI - Study of the immunostimulating effect of glycophosphopeptical (AM3) in mice. AB - The results of this study show that glycophosphopeptical (AM-3) has a marked immunostimulant effect in mice, in terms of an increase in the number of haemolytic plaque-forming B lymphocytes producing antibodies against sheep erythrocytes as compared with saline-treated controls. The results also demonstrate the activity of this drug when administered intraperitoneally. PMID- 9215592 TI - Comparing long-term depression with pharmacologically induced synaptic attenuations in young rat hippocampi. AB - Field excitatory postsynaptic potentials (EPSPs) were recorded in the CA1 region of hippocampal slices from 12-18-day-old rats. The isolated N-methyl-D-aspartate (NMDA) receptor mediated field EPSP as well as the composite field EPSP of both NMDA and alpha-amino-3-hydroxy-5-methylisoxazolepropionic acid (AMPA) receptor mediated components were obtained in low Mg2+ solutions with 10 microM or 1 microM of the AMPA receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), respectively. The isolated AMPA receptor mediated field EPSP was obtained either in normal Mg2+ solution or in a low Mg2+ solution in the presence of the NMDA receptor antagonist D-2-amino-5-phosphonopentanoic acid. The waveforms of the field EPSPs were studied and the effect of long-term depression (LTD) on these waveforms was compared with the effects of several pharmacological agents that attenuate the synaptic efficacy. It was shown that LTD occurred without changes in the waveforms of isolated AMPA and NMDA EPSPs. Reducing the number of release sites by lowering the stimulus strength or reducing the probability of transmitter release by an adenosine agonist N6-cyclohexyl-adenosine both mimicked the LTD-induced changes. Partial blockade of the AMPA receptors was also without effect on the waveforms of isolated AMPA EPSPs. In contrast, partial blockade of the NMDA receptors in several different ways resulted in waveform changes. A similar result could be inferred from experiments using composite field EPSPs. The synaptic attenuation caused by a partial blockade of NMDA receptors therefore appears to differ mechanistically from that involved in LTD, arguing against a postsynaptic locus of the modification involved in LTD. However, directly testing for alterations in transmitter release using the open channel blocker of NMDA receptors MK-801 failed in revealing such presynaptic changes during LTD. Our results therefore suggest that LTD might be due to a coordinated pre- and postsynaptic change instead of distinct pre- or postsynaptic modifications. PMID- 9215593 TI - Autoradiographic distribution of M1, M2, M3, and M4 muscarinic receptor subtypes in Alzheimer's disease. AB - We studied the autoradiographic densities of all pharmacologically characterised muscarinic receptors (MR) in frontal, temporal, and visual cortex, hippocampal formation, and striatum in autopsied brains from 19 histopathologically verified patients of Alzheimer's disease (AD) and in matched controls. Almost all (16 of 19) of the AD cases were severe. In AD brains, total MR, M1, and M3 MR subtypes were found to be significantly decreased in entorhinal cortex and in most hippocampal strata. Total MR and M1 receptors were also significantly reduced in visual area and in frontal cortex of AD brains, respectively. M2 receptors were significantly reduced over hippocampal formation but increased significantly in striatum of AD brains as compared with controls. M3 receptors in AD were in the range of controls in neocortex and striatum, whereas the M4 receptor subtype was also preserved in all brain regions in AD brains when compared with controls. This is the first autoradiographic study analysing the distribution of all MR subtypes in AD brains. These changes in MR densities concur with the general pattern of neuronal degeneration occurring in AD brains and partly explain the poor response of AD cognitive decline to present cholinergic supplementation therapies. Although M3 and M4 MR were labelled with nonselective approaches, the preservation of M4 and to a lesser degree M3 MR subtypes in AD brains could open an alternative way for the symptomatic therapy of AD dementia. PMID- 9215594 TI - Enhancement of basal and pentylenetetrazol (PTZ)-stimulated dopamine release in the brain of freely moving rats by PTZ-induced kindling. AB - The effects of pentylenetetrazol (PTZ)-induced kindling on the activity of mesocortical, mesoaccumbens, and nigrostriatal dopaminergic neurons was investigated with the transversal microdialysis technique in freely moving rats. Four days after the last chronic administration of PTZ, the basal extracellular concentrations of dopamine in the prefrontal cortex, nucleus accumbens, and striatum of kindled rats were significantly increased (+76, +36, +49%, respectively) relative to those of animals chronically treated with saline. Moreover, dopamine output was markedly more sensitive to the effect of a challenge injection of PTZ (20 mg/kg ip) in the prefrontal cortex (+93 vs. +50%, relative to basal values), the nucleus accumbens (+36 vs. +4%), and the striatum (+50 vs. + 35%) of kindled rats relative to that in the control animals. Because kindled rats and their controls are habituated to handling, the neurochemical mechanisms that underlie the effects of chemical kindling on the sensitivity of dopaminergic neurons to PTZ were investigated by comparing the effects of an acute administration of PTZ (20 mg/kg ip) between naive and handling-habituated animals. The sensitivity of dopamine output to PTZ in naive rats was markedly greater than that in handling-habituated animals for the prefrontal cortex (+83 vs. +50%) and nucleus accumbens (+35 vs. +4%), but not for the striatum (+35 vs. +32%). These results indicate that PTZ kindling enhances the basal activity and the sensitivity to PTZ of dopamine neurons in rat brain and suggest that mesocortical, mesoaccumbens, and nigrostriatal dopaminergic neurons contribute to the central alterations associated with experimental epilepsy. PMID- 9215595 TI - Effects of drugs interfering with sodium channels and calcium channels on the release of endogenous dopamine from superfused substantia nigra slices. AB - The importance of voltage-dependent sodium channels and different types of voltage-sensitive calcium channels for depolarisation-induced release of endogenous dopamine from dendrites and cell bodies in superfused guinea pig substantia nigra slices was investigated. The stimulatory effect of veratridine (10 microM) on dopamine release was only marginally attenuated in Ca(2+)-free medium but was completely blocked by tetrodotoxin (1 microM) and by the dopamine reuptake inhibitor GBR 12909 (10 microM). Low extracellular concentration of Na+ stimulated the dopamine release. Potassium-evoked dopamine release was completely Ca(2+)-dependent, not blocked by GBR 12909 and partially blocked by tetrodotoxin. Nifedipine (20 microM), omega-conotoxin GVIA (0.5 microM), penfluridol (5 microM), and Ni2+ (20 microM) had no effect, amiloride (1 mM) attenuated and neomycin (350 microM), and omega-agatoxin IVA (1 microM) almost totally blocked the potassium-induced dopamine release. The results suggest that veratridine released dopamine mostly by reversing the dopamine transporter. High concentrations of potassium induced release of nigral dopamine by opening of voltage-sensitive calcium channels of P/Q type but not L-type, N-type and probably not T-type. The depolarisation evoked by high concentrations of potassium seems to open voltage-sensitive calcium channels both by the depolarisation induced by potassium per se and by the secondary depolarisation induced by opening of voltage-dependent sodium channels. PMID- 9215596 TI - Involvement of cortical neurotensin in the regulation of rat meso-cortico-limbic dopamine neurons: evidence from changes in the number of spontaneously active A10 cells after neurotensin receptor blockade. AB - In order to further assess the role of endogenous neurotensin on midbrain dopaminergic neuronal function, the effects of the selective neurotensin receptor antagonists SR 48692 and SR 48527 were investigated on the number of spontaneously active A9 and A10 dopaminergic neurons in rats. Single intraperitoneal administration of SR 48692 (0.1-3 mg/kg) dose-dependently increased the number of active A10, but not A9 cells. SR 48527 (1 mg/kg) had a similar profile, but not SR 49711, its low affinity R-enantiomer, indicating that the effects observed were mediated through neurotensin receptor blockade. Five week treatment with SR 48692 (3 mg/kg/day) produced a significant decrease of the number of active A10, but not A9 cells, which was reversed by apomorphine, suggesting that these cells were under depolarization block. Single co administration of inactive doses of SR 48692 (0.1 mg/kg) and haloperidol (0.0625 mg/kg) significantly increased the number of active A10 cells. Conversely, co administered active doses of SR 48692 or SR 48527 and haloperidol (1 and 0.25 mg/kg, respectively) induced an apomorphine-sensitive decrease of the number of A10 active cells. Finally, SR 48692 (10 mg/kg) modified neither accumbal nor cortical basal DA release. Local micro-injection of SR 48692 (10[-11]-10[-9] M), but not that of SR 49711 (10[-9] M), into the prefrontal cortex, increased the number of active A10 cells in a concentration-dependent manner. These results suggest that neurotensin receptor blockade counteracts a tonic inhibitory regulation by endogenous neurotensin of mesolimbic dopaminergic function and indicate that the prefrontal cortex is critically involved in this regulation. PMID- 9215598 TI - [125/123I]IPH: a radioiodinated analog of epibatidine for in vivo studies of nicotinic acetylcholine receptors. AB - Tomographic imaging of central nicotinic acetylcholine receptors (nAChRs) via single photon emission computed tomography (SPECT) has been hampered by the lack of a radioligand with suitable in vivo binding characteristics. Therefore, a novel analog of epibatidine, (+/-)-exo-2-(2-iodo-5-pyridyl)-7 azabicyclo[2.2.1]heptane (IPH), labeled with [(125)I] or [(123)I] was evaluated as an in vivo marker of central nicotinic acetylcholine receptors (nAChRs). [(125)I]IPH showed substantial brain penetration (4.2% of the injected dose at 30 min) and a cerebral biodistribution in mice consistent with the in vivo labeling of nAChRs (% injected dose/gram of thalamus, superior colliculi >> cerebellum). [(125)I]IPH binding sites were shown to be saturable with unlabeled IPH (ED50 approximately 1 microg/kg). The uptake of [(125)I]IPH was blocked significantly by the nicotinic agonists, cytisine, lobeline, and (-)-nicotine, but not by the noncompetitive nAChR antagonist, mecamylamine. Antagonists of muscarinic (scopolamine), serotonin (ketanserin), and opioid (naloxone) receptors had no significant effect on [(125)I]IPH binding. A preliminary SPECT imaging study with [(123)I]IPH in a baboon showed [(123)I]IPH to localize in nAChR-rich areas of brain (thalamus > frontal cortex > cerebellum). [(123)I]IPH binding in baboon brain was also displaced (35-45% displacement) by a challenge dose of cytisine showing that a well-characterized nicotinic agonist effectively competes for [(123)I]IPH binding sites. [(123)I]IPH seems well suited for imaging studies of nAChRs and, to our knowledge, is the first SPECT agent that has allowed for the visualization of nAChRs in primate brain. PMID- 9215597 TI - Differential interaction of competitive NMDA and AMPA antagonists with selective dopamine D-1 and D-2 agonists in a rat model of Parkinson's disease. AB - Stimulation of the dopamine (DA) D-2 and D-1 receptors results in behavioural activation (i.e., induction of contralateral rotations) in 6-hydroxydopamine (6 OHDA) substantia nigra lesioned rats. Competitive N-methyl-D-aspartate (NMDA) antagonists as well as alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) antagonists potentiate the stimulatory responses to threshold doses of L DOPA or the mixed dopamine D-1/D-2 agonist apomorphine in this model, indicating the potential of such combinations for the management of Parkinson's disease. Neuroanatomic and electrophysiologic data indicate a differential distribution of DA D-1 and DA D-2 receptors within motor loops of the basal ganglia. DA D-1 receptors are preferentially located on GABAergic neurones projecting to the substantia nigra compacta (SNc) and to the substantia nigra reticulata (SNr), whereas DA D-2 receptors are preferentially located on neurones that innervate the external pallidum. NMDA receptors are present in high densities within the striatum, whereas AMPA receptors are enriched in the entopeduncular nucleus/internal pallidum and the SNr. To further characterise the functional interaction between DA and glutamate receptors, we tested the competitive NMDA antagonist 3-((+/-)-2-carboxypiperazin-4-yl)-propyl-1-phosphonic acid (CPP) and the AMPA antagonist 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo[f] quinoxaline (NBQX) following systemic administration in combination with the DA D-2 selective agonist quinpirole or the DAD-1 selective agonist A 68 930 (1R,3S)-1-aminomethyl 5,6-dihydroxy-3-phenylisochroman) in rats with chronic 6-OHDA lesions of the SNc. CPP potentiated quinpirole-induced rotations and did not affect those induced by the D-1 agonist A 68930. By contrast, NBQX had no effect on quinpirole-induced rotations, whereas synergism was seen with A 68930. These results suggest that rotations induced by combined treatment with glutamate antagonists and DA agonists are mediated by different pathways within the basal ganglia, depending on which subtype of receptor is involved. AMPA antagonists could act preferentially by activating the direct motor pathway, whereas NMDA antagonists could modulate the indirect loop. PMID- 9215599 TI - Coexpression of striatal dopamine receptor subtypes and excitatory amino acid subunits. AB - The striatal cellular coexpression patterns for the D(1A) and D2 dopamine (DA) receptor subtypes and the ionotropic excitatory amino acid (EAA) subunits of the N-methyl-D-aspartate (NMDA-R1) and the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) (GluR1 and GluR2/3) receptor subunits were examined morphologically. Their coincidence was assessed by visualization of mRNA transcripts, localization of encoded receptor proteins, and binding analysis using concurrently paired methods of fluorescence detection. The findings indicated that 1) mRNA transcripts for both receptor systems were detected in the medium-sized neuron population, and the distribution of receptor message closely reflected protein and binding patterns, with the exception of the GluR1 subunit; 2) both DA receptor mRNA transcripts were coexpressed with each ionotropic EAA receptor subunit examined and with each other, and NMDA and AMPA receptor subunits also showed coincident expression; 3) D(1A) DA receptor protein was detected in neurons which coexpressed EAA subunit proteins; and 4) GluR2/3 and NMDA-R1 subunit proteins were coexpressed in medium-sized neurons which also demonstrated D2 DA receptor binding sites. These findings suggest morphological receptor "promiscuity" since the coexpression patterns between DA and EAA receptors were found in all permutations. The results provide a spatial framework for physiological findings describing functional interactions between the two DA receptor types and between specific DA and EAA receptors in the striatum. PMID- 9215600 TI - Characterization of dopamine-induced depolarization of prefrontal cortical neurons. AB - Dopamine (DA) has been reported to depolarize neurons in the prefrontal cortex (PFC). To further characterize this effect of DA, we made whole cell recordings from PFC pyramidal cells in rat brain slices. As reported previously, DA depolarized most PFC cells tested. This effect of DA was concentration-dependent and persisted in the presence of synaptic blockade, indicating a direct effect of DA on the recorded cell. During DA-induced depolarization, PFC neurons consistently showed an increase in excitability, suggesting that the depolarization is not directly related to DA-induced inhibition of PFC neurons previously observed in vivo. Surprisingly, the effect of DA was not mimicked or blocked by several commonly used DA agonists and DA antagonists. The alpha and beta antagonists phentolamine and alprenolol and the atypical antipsychotic drug clozapine also showed no significant effect on DA-induced depolarization. These results suggest that DA-induced depolarization may be mediated by a nonspecific mechanism. However, it remains possible that there exists a new type of DA receptors in the PFC not sensitive to classical DA agonists and antagonists, particularly given the fact that DA applied in the same manner depolarized only PFC neurons but not those in the striatum or the substantia nigra. PMID- 9215601 TI - Direct approach for attenuating cocaine's effects on extracellular dopamine: targeting the dopamine transporter. AB - Using in vivo microdialysis techniques, the effects of RTI-55 and/or cocaine on extracellular dopamine (DA) concentrations were measured in the nucleus accumbens (NACC) of freely moving rats. In control animals, cocaine (20 mg/kg) increased NACC DA approximately 458% 60 minutes following administration, returning to baseline values within 200 minutes. Similarly, RTI-55 administration (0.25 mg/kg) increased NACC DA levels approximately 347%. When combined, however, cocaine further increased NACC DA to 705% of baseline values when given 4 hours following RTI-55. This increase was significantly larger than cocaine alone (P < 0.05). In addition, chronic RTI-55 administration (5 days) further potentiated cocaine's ability to increase NACC DA (783%) but this did not reach statistical significance (P > 0.1) compared to acute RTI55/cocaine animals. These findings indicate that RTI-55, a drug that binds directly to the dopamine transporter (DAT) with higher affinity than cocaine, does not appear to be effective in attenuating cocaine's effects on NACC dopamine levels. In fact, acute RTI-55 potentiates cocaine's effects on NACC DA. PMID- 9215602 TI - Fostriecin: a review of the preclinical data. AB - Fostriecin is a novel antitumor antibiotic. In vitro studies showed that fostriecin inhibits DNA topoisomerase II (Topo II) catalytic activity, protein phosphatases involved with cell-cycle control and histone phosphatases. The relative contribution of these mechanisms to the antitumor activity has not been elucidated, but Topo II inhibition seems to be the major mechanism of action at in vitro cytotoxic fostriecin levels. Tumor cell lines with decreased Topo II content showed similar or increased sensitivity to fostriecin, compared to the parent cell lines. The reduced-folate carrier is probably responsible for the cellular uptake of fostriecin. The possible clinical consequences of these in vitro observations are discussed. PMID- 9215603 TI - Trofosfamide: a review of its pharmacodynamic and pharmacokinetic properties and therapeutic potential in the oral treatment of cancer. AB - During the last decade, the oxazaphosphorine trofosfamide was underestimated partly due to its unsuitability for i.v. use. Oral daily doses of 150 mg were tolerated well and showed appreciable response rates in the treatment of lymphoma. Furthermore, activity in sarcoma and cancers sensitive to oxazaphosphorines in general seems probable, because ifosfamide is the main metabolite of trofosfamide. Due to its oral mode of application and good tolerance, trofosfamide will be an important option in view of the increasing demand for treatment regimens suited for an outpatient basis. Results of the major in vitro, in vivo and clinical studies are reported for evaluation of its significance in chemotherapy today. PMID- 9215604 TI - Efficacy of carboplatin plus primary prophylactic filgrastim (granulocyte colony stimulating factor) in relapsed ovarian cancer: a study of the Gynecologic Oncology Group of the Comprehensive Cancer Center Limburg. AB - A total of 34 patients with advanced ovarian cancer, who relapsed 1-72 months after at least one first-line cisplatin-based chemotherapy protocol, were treated with carboplatin, 350 mg/m2 q 4 weeks, with the adjunct of primary prophylactic granulocyte colony stimulating factor (G-CSF; filgrastim), 300 or 480 microg daily, days 5-9. Over 90% of the anticipated dose of carboplatin could be administered. Partial response, defined as a decline in CA-125 of 50% or more on two consecutive samples, occurred in 42%, while 15% of patients achieved a complete response (no clinical signs of disease with normalization of CA-125). Survival from start of carboplatin treatment was 23 months. Myelosuppression was the most important toxicity with 35% of patients experiencing grade 4 thrombocytopenia of short duration. Grade 4 leucopenia occurred in only one patient. It is concluded that single-agent carboplatin, with the adjunct of prophylactic G-CSF, can be administered with adequate dose intensity, and is an effective and acceptable palliative treatment for patients with relapse after first-line cisplatin-based chemotherapy. PMID- 9215605 TI - Comparison of oral itasetron with oral ondansetron: results of a double-blind, active-controlled phase II study in chemotherapy-naive patients receiving moderately emetogenic chemotherapy. AB - Itasetron hydrochloride is a new 5-hydroxytryptamine3 (5-HT3) antagonist. Experimental investigations show that orally it is rapidly absorbed (about 90 min), is highly bioavailable (greater than 90%), has a long half-life (about 12 h) and is more potent (about 10 times) in animal models than ondansetron, currently standard therapy for the prophylactic control of chemotherapy induced nausea and vomiting. This paper describes the results of a study designed to assess the efficacy and tolerability of five (0.5, 1, 2, 4 and 8 mg) twice-daily doses of itasetron hydrochloride, in comparison with 8 mg b.i.d. ondansetron. Assessments were made in patients (n = 104) with histologically confirmed cancer (excluding head and neck tumors) and about to receive their first course of moderately emetogenic chemotherapy. Itasetron hydrochloride demonstrated comparable efficacy to ondansetron; no statistically significant between-group differences were observed in the primary (complete response rate) or secondary (nausea and delayed emesis) efficacy criteria. Adverse events were similar in type and incidence across all treatment groups, and were those expected for this therapeutic class. The tolerability of itasetron hydrochloride was assessed as 'very good' or 'rather good' by 81% of patients and 89% of physicians. In conclusion, itasetron hydrochloride is effective and well tolerated in patients receiving moderately emetogenic chemotherapy. Oral doses of 1 mg b.i.d. or above will be used in further clinical studies. PMID- 9215606 TI - Stability and analysis of 2-chloro-2'-deoxyadenosine, 2-chloro-2'-arabino-fluoro 2'-deoxyadenosine and 2-chloroadenine in human blood plasma. AB - Cladribine (2-chloro-2'-deoxyadenosine, CdA) is a purine nucleoside analog with activity against lymphoproliferative and autoimmune disorders. 2-Chloro-2' arabino-fluoro-2'-deoxyadenosine (CAFdA), a derivative of CdA with better acid stability, shows a similar in vitro spectrum of activity as CdA. 2-Chloroadenine (CAde) is the major catabolite of both CdA and CAFdA. We have developed a high performance liquid chromatography method to measure CdA, CAFdA and their metabolite CAde in plasma. This method employees an internal standard, chloroadenosine (CAdo), and a C8 solid-phase extraction to isolate and concentrate the substances. Chromatographic separation was achieved using a C8 reverse-phase column, with UV detection at 265 nm, which gives a limit of detection of 1 nmol/l for all substances. The method was reproducible with intra- and inter-assay coefficients of variations below 6% at 50 nmol/l and at 5 nmol/l below 23%. The average recoveries of CdA, CAde, CAFdA and the internal standard were higher than 70%. Stability studies of authentic patient samples show that samples containing CdA should be kept in a refrigerator or on ice to prevent degradation. Plasma containing CAde should not be kept at -20 degrees C for longer than 10 weeks before analysis. CdA and CAFdA remain almost stable during storage at -20 degrees C for 12 weeks. PMID- 9215607 TI - Weekly chronomodulated 48 h infusion of high-dose 5-fluorouracil modulated by methotrexate and (6S)-leucovorin in advanced colorectal cancer: a phase IB study. AB - In this phase IB study, 24 patients with advanced colorectal cancer were treated with escalating doses of weekly chronomodulated 48 h infusions of 5-fluorouracil (5-FU) biochemically modulated by methotrexate 40 mg/m2 and (6S)-leucovorin 8 x 45 mg orally. Two daily peak delivery periods (PDP), during which 65% of the daily dose was administered, were investigated: from 18.00 to 0.30 h and from 0.00 to 06.30 h. The maximal tolerated dose of 5-FU was 2800 mg/m2/48 h, with a PDP from 18.00 to 0.30 h. PMID- 9215608 TI - Effect of medroxyprogesterone acetate on the quality of life of the oncologic patient: a multicentric cooperative study. AB - Anorexia and cachexia, major problems in patients with cancer, lead to decreased caloric intake and weight loss. Successful treatment of these conditions has a positive effect on patients' quality of life. Among the pharmacologic treatments, partial effects have been observed following administration of corticosteroids, anabolizing drugs and synthetic progestogens such as megestrol acetate and medroxyprogesterone acetate (MPA). The aim of the present study was to evaluate whether MPA is able to influence the quality of life of neoplastic patients undergoing different chemotherapeutic regimens and/or radiotherapy for different tumor types. A series of 279 cancer patients undergoing either chemotherapy and/or radiotherapy treatment for different tumor types was randomly allocated to receive either MPA or no treatment. We explored the effect of MPA oral suspension at the daily dose of 1000 mg for 12 weeks (group A) or no treatment (group B). Our data show an increase of body weight in group A patients and improvement in performance status. The outcome of the present study strongly demonstrates that therapy with MPA plays a fundamental role in ameliorating the complex symptomatology of cancer patients in intermediate or advanced stage of the disease undergoing casual treatment with chemotherapy and/or radiotherapy. PMID- 9215609 TI - Effect of low-dose 1-hydroxyvitamin D3 in a patient with myelodysplasia after induction therapy for acute myelocytic leukemia. PMID- 9215610 TI - Severe adverse skin reaction to chlorambucil in a patient with chronic lymphocytic leukemia. PMID- 9215611 TI - Antiproliferative effect of curcumin (diferuloylmethane) against human breast tumor cell lines. AB - Pharmacologically safe compounds that can inhibit the proliferation of tumor cells have potential as anticancer agents. Curcumin, a diferuloylmethane, is a major active component of the food flavor turmeric (Curcuma longa) that exhibits anticarcinogenic properties in vivo. In vitro, it suppressed c-jun/Ap-1 and NF kappaB activation and type 1 human immunodeficiency virus long-terminal repeat directed gene expression. We examined the antiproliferative effects of curcumin against several breast tumor cell lines, including hormone-dependent and independent and multidrug-resistant (MDR) lines. Cell growth inhibition was monitored by [3H]thymidine incorporation, Trypan blue exclusion, crystal violet dye uptake and flow cytometry. All the cell lines tested, including the MDR positive ones, were highly sensitive to curcumin. The growth inhibitory effect of curcumin was time- and dose-dependent, and correlated with its inhibition of ornithine decarboxylase activity. Curcumin preferentially arrested cells in the G2/S phase of the cell cycle. Curcumin-induced cell death was neither due to apoptosis nor to any significant change in the expression of apoptosis-related genes, including Bcl-2, p53, cyclin B and transglutaminase. Overall our results suggest that curcumin is a potent antiproliferative agent for breast tumor cells and may have potential as an anticancer agent. PMID- 9215612 TI - Irreversible cytotoxic effect of a novel lowly immunosuppressive antitumor fluorouridine derivative, UK-21. AB - Previously, we reported that the 5-fluorouridine derivative, 2',3',5'-tris-O-[N(2 n-propyl-n-pentanoylglycyl]-5-fluorouridine (UK-21), is a newly synthesized lowly immunosuppressive and potent antitumor drug in comparison with other fluorouridine derivatives such as 5-fluorouracil (5-FU), 5-fluorouridine (5-FUR) and 5-fluorodeoxyuridine (5-FUDR). In order to elucidate the molecular mechanism of antitumor activity of UK-21, we compared the effect of the four drugs on cell proliferation, cell cycle progression and macromolecular syntheses. When KB cells were subjected to a colony-forming inhibition assay designed to expose the cells to the drugs for 4-96 h and wash out, UK-21 and 5-FUR inhibited the colony formation at concentrations ranging from 0.01 to 0.1 microM, whereas 1-100 microM was needed for the cytotoxicity of 5-FU and 5-FUDR. By exposure for 24-48 h, all these drugs inhibited cell growth and caused accumulation of the cells in S or G2 phase at almost the same concentrations of 0.32-8 microM. These results suggest that the cytotoxic effects of UK-21 and 5-FUR are irreversible, while those of 5 FU and 5-FUDR are reversible. To confirm this, KB cells were treated with UK-21 and/or 5-FU for 1 h, and continued to be cultured for 1-7 days, resulting in the inhibition of the cell growth by UK-21 in a dose-dependent manner at concentrations of 10-100 microM, but not by 5-FU even at 100 microM. UK-21, 5-FUR and 5-FU showed a linear relationship between exposure time and IC50 in the colony formation assay with a slope of almost -1, but 5-FUDR did not, suggesting that UK-21, 5-FUR and 5-FU are cell cycle non-specific inhibitors, while 5-FUDR is a cell cycle-specific inhibitor. UK-21 and 5-FUR, but not 5-FU and 5-FUDR inhibited the incorporation of [3H]uridine into the acid insoluble fraction, while UK-21 and 5-FUDR, but not 5-FUR and 5-FU inhibited the incorporation of [3H]thymidine. These results suggest that irreversible cytotoxic effects of UK-21 like 5-FUR are exerted through inhibition of RNA synthesis. PMID- 9215613 TI - Synthesis and cytotoxic activity of indolyl thiazoles. AB - A number of indolyl thiazoles have been prepared and evaluated for their antitumor properties. The compounds were synthesized from the appropriate indole, building up the thiazole ring using the Hantzsch reaction. Cytotoxic activity was measured in the human breast cancer cell line SKBr3 using the MTT assay. PMID- 9215614 TI - A new synthetic lipid A analog, ONO-4007, stimulates the production of tumor necrosis factor-alpha in tumor tissues, resulting in the rejection of transplanted rat hepatoma cells. AB - ONO-4007 is a new synthetic lipid A derivative with low endotoxic activities. We have examined the therapeutic effects of ONO-4007 on rat hepatocellular carcinoma KDH-8 cells, rat fibrosarcoma KMT-17 cells and rat mammary adenocarcinoma SST-2 cells in vivo. Multiple systemic i.v. administration of ONO-4007 was performed on days 7, 14 and 21 after tumor implantation of KDH-8 and SST-2 cells, and on days 5, 10 and 15 after tumor implantation of KMT-17 cells. ONO-4007 showed significant therapeutic effects on KDH-8 cells; by the administration of ONO-4007 (2.5 mg/kg) 70% of rats were cured and by the administration of ONO-4007 (5 mg/kg) 50% of rats were cured. Furthermore, the ONO-4007 treatment prolonged the mean survival time of KDH-8-bearing rats. However, ONO-4007 had no effect on KMT 17 and SST-2 cells, and it had no direct effect on the growth of KDH-8 cells in vivo. Albeit the stimulation with ONO-4007 induced mRNA expressions of interleukin (IL)-1alpha, IL-6 and tumor necrosis factor (TNF)-a, those of IL-2, IL-4, IL-10 and interferon (IFN)-gamma were not induced. Using a bioassay, we found that the production of TNF-alpha in the tumor tissues was induced by ONO 4007 in a dose-dependent manner. KDH-8 cells were sensitive to human natural TNF alpha in vitro. However, KMT-17 and SST-2 cells were resistant against TNF-alpha in vitro. These results suggest that ONO-4007 is therapeutically useful for the treatment of TNF-alpha-sensitive tumors. PMID- 9215616 TI - Deoxycytidine kinase mRNA expression in childhood acute lymphoblastic leukemia. AB - Initial and relapsed childhood acute lymphoblastic leukemias (ALL) were investigated for mutations and expression of deoxycytidine kinase (dCK). dCK mediating toxicity of 1-beta-D-arabinofuranosylcytosine was analyzed semiquantitatively by RT-PCR and by single-strand conformation polymorphism. We found that patients with initial ALL experienced more frequently a relapse if dCK expression was low or absent. No mutations were found in initial and relapsed ALL. PMID- 9215615 TI - Modulation of melphalan and cisplatin cytotoxicity in human ovarian cancer cells resistant to alkylating drugs. AB - We investigated the effect of pharmacological modulators on the cytotoxic activity of melphalan and cisplatin in human ovarian cystadenocarcinoma cells sensitive (OAW42) or resistant (OAW42MER) to bifunctional alkylating agents. By filter elution experiments we observed a reduced accumulation and a faster repair of melphalan-induced DNA interstrand cross-links in the OAW42MER resistant cells than in the OAW42 parental, sensitive cells. Moreover, resistant cells were characterized by an increased level of mRNA encoding enzymes involved in the nucleotide excision repair pathway, such as ERCC (excision repair cross complementing)1 and ERCC2. Among the modulators used, the topoisomerase I inhibitor topotecan was able to increase melphalan cytotoxic activity in sensitive and resistant cell lines. Topotecan also positively modulated cisplatin activity, although to a variable extent in the two cell lines, as a function of treatment schedule. The energolytic compound lonidamine markedly enhanced the cytotoxicity of melphalan and cisplatin, with a potentiating effect in the OAW42MER resistant cells almost 2-fold that of in the OAW42 sensitive cells. No significant potentiation was observed by using calcium channel blockers, such as verapamil and nimodipine. Conversely, an increase in melphalan cytotoxic activity was determined by flunarizine in OAW42MER resistant cells and, to a lesser extent, in OAW42 sensitive cells. However, the calcium blocker failed to modulate cisplatin activity in both cell lines. PMID- 9215617 TI - The additive antiproliferative effect of all-trans retinoic acid and interferon alpha2a on human cervical carcinoma cell lines is not associated with increased expression of retinoid receptors. AB - The ability of all-trans retinoic acid (atRA), interferon-alpha2a (IFN-alpha2a) or a combination thereof to modulate the growth of three human cervical carcinoma cell lines (ME180, MS751 and CaSki) and the relationship between responsiveness and the expression of cytosolic retinoid-binding proteins (CRBP and CRABP II), nuclear RA receptors (RAR-alpha, -beta and -gamma) and retinoid X receptor (RXR alpha) were investigated. atRA induced an antiproliferative effect on two of the cell lines (ME180 > MS751), whereas CaSki was much less responsive. An additive growth inhibition on the latter two cell lines was achieved with the combined treatment of atRA and IFN-alpha2a. Receptor expression appeared to be unrelated to growth inhibition in these cell lines in so far as atRA exerted minimal effect on the growth of CaSki, although these cells expressed four of these nuclear receptors. However, mRNA for CRABP II was not demonstrable in CaSki, in contrast to the other two atRA responsive cell lines, as evaluated with RT-PCR and ethidium bromide staining. Treatment with atRA or IFN-alpha2a did not induce any change in mRNA for the nuclear retinoid receptors or cellular retinoid binding proteins after 3 or 6 days of treatment. PMID- 9215618 TI - Mitotic arrest and anaphase aberrations induced by vinorelbine in hamster cells in vitro. AB - Aneugenic effects of the Vinca alkaloid vinorelbine (VRB) were evaluated in vitro, measuring sister chromatid exchanges (SCE), cell proliferation kinetics and anaphase telophase aberrations in Chinese hamster ovary (CHO) cells. The highest dose of VRB (0.50 microg/ml) arrested cells at the first metaphase. An increase in abnormal anaphases was seen at 0.05-0.50 microg/ml of VRB, containing chiefly lagging chromosomes and multipolar spindles. No increase in SCE was found. These results indicate that VRB does not directly damage DNA, but acts on spindle microtubules, altering chromosome movement and causing aneuploidy. PMID- 9215619 TI - Archaeal genomics: an overview. PMID- 9215620 TI - Archaea and the origin(s) of DNA replication proteins. PMID- 9215621 TI - Archaeal histones, nucleosomes, and transcription initiation. PMID- 9215622 TI - Another bridge between kingdoms: tRNA splicing in archaea and eukaryotes. PMID- 9215623 TI - Ancient ciphers: translation in Archaea. PMID- 9215624 TI - Bcl-2 can rescue T lymphocyte development in interleukin-7 receptor-deficient mice but not in mutant rag-1-/- mice. AB - Signals from cytokine and antigen receptors play crucial roles during lymphocyte development. Mice lacking interleukin-7 receptor are lymphopenic, due to a defect in cell expansion at an early stage of differentiation, and the few mature T cells that develop in IL-7R-/- animals are functionally impaired. Both defects were rescued completely by overexpression of the anti-apoptosis protein Bcl-2. T cell progenitors lacking antigen receptor molecules are also blocked in differentiation and die, presumably because they fail to receive a positive signal via their pre-T cell receptor. Surprisingly, Bcl-2 did not promote survival or differentiation of T cells in rag-1-/- mice. These results provide evidence that blocking apoptosis is the essential function of IL-7R during differentiation and activation of T lymphocytes and that pre-TCR signaling blocks a pathway to apoptosis that is insensitive to Bcl-2. PMID- 9215625 TI - Enforced expression of Bcl-2 in monocytes rescues macrophages and partially reverses osteopetrosis in op/op mice. AB - Osteopetrotic (op/op) mice lack functional M-CSF and have depressed levels of macrophages and osteoclasts. We prepared transgenic mice (hMRP8bcl-2) that express human Bcl-2 in monocytes. In vitro hMRP8bcl-2 monocytes do not undergo apoptosis in the absence of serum and M-CSF, while op/op and wild-type monocytes die. These Bcl-2-expressing monocytes spontaneously undergo macrophage differentiation. In vivo, the op/op hMRP8bcl-2 mice show significant replenishment of tissue macrophages. Their long bone osteopetrosis is largely reversed, and extensive medullary hematopoiesis appears in the bone marrow. We propose that M-CSF augments monocyte survival, permitting them to respond to internal and external cues for their differentiation. PMID- 9215626 TI - Bcl-2 rescues T lymphopoiesis in interleukin-7 receptor-deficient mice. AB - Mice lacking functional IL-7 or IL-7R alpha genes are severely deficient in developing thymocytes, T cells, and B cells. IL-7 and IL-7 receptor functions are believed to result in lymphoid cell proliferation and cell maturation, implying signal transduction pathways directly involved in mitogenesis and elaboration of developmentally specific new gene programs. Here, we show that enforced expression of the bcl-2 gene in T-lymphoid cells (by crossing in the Emu-bcl-2 transgene) in IL-7R alpha-deficient mice results in a significant restoration of thymic positive selection and T cell numbers and function. We propose cell survival signals to be the principal function of IL-7R engagement in thymic and T cell development. PMID- 9215627 TI - Proteolysis that is inhibited by hedgehog targets Cubitus interruptus protein to the nucleus and converts it to a repressor. AB - Cell-cell communication at anterior/posterior compartment borders in Drosophila involves Hedgehog (Hh), a protein secreted by posterior cells, and Cubitus interruptus (Ci), a protein in the Hh response pathway in anterior cells. Although Ci is thought to have roles as a transcription factor repressing hh expression and activating target genes, it localizes in the cytoplasm of anterior cells. We report here the identification of a domain that tethers Ci in the cytoplasm and show that in some anterior cells, Ci is cleaved to generate a form that lacks the tethering domain. This form translocates to the nucleus where it represses hh and other target genes. Hh inhibits proteolysis of Ci, and we suggest that this inhibition leads to the observed patterns of expression of key target genes at the compartment border. PMID- 9215628 TI - 14-3-3 proteins are essential for RAS/MAPK cascade signaling during pseudohyphal development in S. cerevisiae. AB - 14-3-3 proteins are highly conserved ubiquitous proteins whose explicit functions have remained elusive. Here, we show that the S. cerevisiae 14-3-3 homologs BMH1 and BMH2 are not essential for viability or mating MAPK cascade signaling, but they are essential for pseudohyphal-development MAPK cascade signaling and other processes. Activated alleles of RAS2 and CDC42 induce pseudohyphal development and FG(TyA)-lacZ signaling in Bmh+ strains but not in ste20 (p65PAK) or bmh1 bmh2 mutant strains. Moreover, Bmh1p and Bmh2p associate with Ste20p in vivo. Three alleles of BMH1 encode proteins defective for FG(TyA)-lacZ signaling and association with Ste20p, yet these alleles complement other 14-3-3 functions. Therefore, the 14-3-3 proteins are specifically required for RAS/MAPK cascade signaling during pseudohyphal development in S. cerevisiae. PMID- 9215629 TI - Daxx, a novel Fas-binding protein that activates JNK and apoptosis. AB - The Fas cell surface receptor induces apoptosis upon receptor oligomerization. We have identified a novel signaling protein, termed Daxx, that binds specifically to the Fas death domain. Overexpression of Daxx enhances Fas-mediated apoptosis and activates the Jun N-terminal kinase (JNK) pathway. A C-terminal portion of Daxx interacts with the Fas death domain, while a different region activates both JNK and apoptosis. The Fas-binding domain of Daxx is a dominant-negative inhibitor of both Fas-induced apoptosis and JNK activation, while the FADD death domain partially inhibits death but not JNK activation. The Daxx apoptotic pathway is sensitive to both Bcl-2 and dominant-negative JNK pathway components and acts cooperatively with the FADD pathway. Thus, Daxx and FADD define two distinct apoptotic pathways downstream of Fas. PMID- 9215630 TI - The yeast spindle pole body is assembled around a central crystal of Spc42p. AB - The spindle pole body (SPB) is the microtubule organizing center (MTOC) in the yeast Saccharomyces that plays a pivotal role in such diverse processes as mitosis, budding, and mating. We have used cryoelectron microscopy and image processing to study the structure of isolated diploid SPBs. We show that SPBs are present in two lateral-size classes, sharing a similar vertical architecture comprised of six major layers. Tomographic reconstructions of heparin-stripped SPBs reveal a central hexagonally packed layer. Overexpression of Spc42p results in the growth of a similar layer, forming a crystal that encircles the SPB. Hence, the SPB is an MTOC that utilizes crystallographic packing of subunits in its construction. PMID- 9215631 TI - Crystal structure of a pol alpha family replication DNA polymerase from bacteriophage RB69. AB - The 2.8 A resolution crystal structure of the bacteriophage RB69 gp43, a member of the eukaryotic pol alpha family of replicative DNA polymerases, shares some similarities with other polymerases but shows many differences. Although its palm domain has the same topology as other polymerases, except rat DNA polymerase beta, one of the three carboxylates required for nucleotidyl transfer is located on a different beta strand. The structures of the fingers and thumb domains are unrelated to all other known polymerase structures. The editing 3'-5' exonuclease domain of gp43 is homologous to that of E. coli DNA polymerase I but lies on the opposite side of the polymerase active site. An extended structure-based alignment of eukaryotic DNA polymerase sequences provides structural insights that should be applicable to most eukaryotic DNA polymerases. PMID- 9215632 TI - Protein-protein communication: structural model of the repression complex formed by CytR and the global regulator CRP. AB - The cAMP receptor protein (CRP) and the LacI-related CytR antiactivator bind cooperatively to adjacent DNA sites at or near promoters, an interaction that involves direct protein contacts. Here, we identify a collection of amino acid substitutions in CytR that reestablish protein-protein communication to mutant CRP proteins specifically defective in cooperative binding with wild-type CytR. To assess the location and spatial arrangement of these substitutions, we built a three-dimensional model of CytR based on the recent X-ray structure of the highly homologous PurR repressor bound to DNA. This approach enables us to specify the patch on CytR's surface that contacts CRP. Furthermore, our results permit the construction of a three-dimensional structure of the higher order nucleoprotein complex formed by CytR and CRP. PMID- 9215633 TI - Plasminogen is required for efficient dissemination of B. burgdorferi in ticks and for enhancement of spirochetemia in mice. AB - The role of the host plasminogen activation system in transmission of and invasion by Borrelia burgdorferi, the tick-borne spirochetal agent of Lyme disease, was investigated using plasminogen (Plg)-knockout mice. PLG was not detected in spirochetes from unfed ticks, but binding occurred as ticks fed on the host's blood. Plasminogen activators were derived from the host blood meal. PLG was required for efficient dissemination of B. burgdorferi within the tick and for enhancement of spirochetemia in mice but was not critical for transmission and infection. These results provide evidence for a bacterium using a vertebrate protease to disseminate in an invertebrate vector and underscores the interplay among vector, pathogen, and host in promoting the life cycle and disease. PMID- 9215634 TI - Regulation of IRK3 inward rectifier K+ channel by m1 acetylcholine receptor and intracellular magnesium. AB - Inward rectifier K+ channels control the cell's membrane potential and neuronal excitability. We report that the IRK3 but not the IRK1 inward rectifier K+ channel activity is inhibited by m1 muscarinic acetylcholine receptor. This m1 modulation cannot be accounted for by protein kinase C, Ca2+, or channel phosphorylation, but can be mimicked by Mg2+. Based on quantitative analyses of IRK3 and two different IRK1 mutant channels bestowed with sensitivity to m1 modulation, we suggest that the resting Mg2+ level causes chronic inhibition of IRK3 channels, and m1 receptor stimulation may lead to an increase of cytoplasmic Mg2+ concentration and further channel inhibition, due to the ability of Mg2+ to lead these channels into a prolonged inactivated state. PMID- 9215635 TI - Activation of the ethylene gas response pathway in Arabidopsis by the nuclear protein ETHYLENE-INSENSITIVE3 and related proteins. AB - Mutations in the Arabidopsis ETHYLENE-INSENSITIVE3 (EIN3) gene severely limit a plant's response to the gaseous hormone ethylene. ein3 mutants show a loss of ethylene-mediated effects including gene expression, the triple response, cell growth inhibition, and accelerated senescence. EIN3 acts downstream of the histidine kinase ethylene receptor, ETR1, and the Raf-like kinase, CTR1. The EIN3 gene encodes a novel nuclear-localized protein that shares sequence similarity, structural features, and genetic function with three EIN3-LIKE (EIL) proteins. In addition to EIN3, EIL1 orEIL2 were able to complement ein3, suggesting their participation in the ethylene signaling pathway. Overexpression of EIN3 or EIL1 in wild-type or ethylene-insensitive2 plants conferred constitutive ethylene phenotypes, indicating their sufficiency for activation of the pathway in the absence of ethylene. PMID- 9215636 TI - Mitochondria are excitable organelles capable of generating and conveying electrical and calcium signals. AB - We report Ca2(+)-induced release of Ca2+ from mitochondria (mCICR) dependent on transitory opening of the permeability transition pore (PTP) operating in a low conductance mode. The Ca2+ fluxes taking place during mCICR are a direct consequence of the mitochondrial depolarization spike (mDPS) caused by PTP opening. Both mDPS and mCICR can propagate from one mitochondrion to another in vitro, generating traveling depolarization and Ca2+ waves. Mitochondria thus appear to be excitable organelles capable of generating and conveying electrical and Ca2+ signals. In living cells, mDPS/mCICR is triggered during IP3-induced Ca2+ mobilization and results in the amplification of the Ca2+ signals primarily emitted from the endoplasmic reticulum. PMID- 9215637 TI - Coassembly of TRP and TRPL produces a distinct store-operated conductance. AB - The Drosophila retinal-specific protein, TRP (transient receptor potential), is the founding member of a family of store-operated channels (SOCs) conserved from C. elegans to humans. In vitro studies indicate that TRP is a SOC, but that the related retinal protein, TRPL, is constitutively active. In the current work, we report that coexpression of TRP and TRPL leads to a store-operated, outwardly rectifying current distinct from that owing to either TRP or TRPL alone. TRP and TRPL interact directly, indicating that the TRP-TRPL-dependent current is mediated by heteromultimeric association between the two subunits. We propose that the light-activated current in photoreceptor cells is produced by a combination of TRP homo- and TRP-TRPL heteromultimers. PMID- 9215638 TI - The MAD-related protein Smad7 associates with the TGFbeta receptor and functions as an antagonist of TGFbeta signaling. AB - TGFbeta signaling is initiated when the type I receptor phosphorylates the MAD related protein, Smad2, on C-terminal serine residues. This leads to Smad2 association with Smad4, translocation to the nucleus, and regulation of transcriptional responses. Here we demonstrate that Smad7 is an inhibitor of TGFbeta signaling. Smad7 prevents TGFbeta-dependent formation of Smad2/Smad4 complexes and inhibits the nuclear accumulation of Smad2. Smad7 interacts stably with the activated TGFbeta type I receptor, thereby blocking the association, phosphorylation, and activation of Smad2. Furthermore, mutations in Smad7 that interfere with receptor binding disrupt its inhibitory activity. These studies thus define a novel function for MAD-related proteins as intracellular antagonists of the type I kinase domain of TGFbeta family receptors. PMID- 9215639 TI - Recruitment of p300/CBP in p53-dependent signal pathways. AB - The products of the p53 and CBP/p300 genes have been individually implicated in control of cell growth and regulation of transcription. p53 is known to act as a positive and negative regulator of gene expression. Here we show that p53, in both wild-type and mutant conformation, forms a specific protein complex with p300. However, in its wild-type but not mutant conformation, p53 inhibits a promoter containing the DNA-binding sequences for the transcription factor AP1, in a p300-dependent manner. p300 stimulates the transcriptional activity of p53 on p53-regulated promoters, and it enhances the responsiveness to a physiological upstream modulator of p53 function, ionizing radiation. A dominant negative form of p300 prevents transcriptional activation by p53, and it counteracts p53 mediated G1 arrest and apoptosis. The data implicate p300 as an important component of p53-signaling, thus providing new insight into the mechanisms of cellular proliferation. PMID- 9215640 TI - Transforming growth factor-beta3 regulates transdifferentiation of medial edge epithelium during palatal fusion and associated degradation of the basement membrane. AB - Studies on transforming growth factor beta3 (TGF-beta3) deficient mice have shown that TGF-beta3 plays a critical role in palatogenesis. These null mutant mice have clefting of the secondary palate, caused by a defect in the process of fusion of the palatal shelves. A critical step in mammalian palatal fusion is removal of the medial edge epithelial cells from the midline seam and formation of continuous mesenchyme. To determine in more detail the role of TGF-beta3 in palatogenesis, we cultured TGF-beta3 null mutant and wild-type control palatal shelves in an organ culture system. The fate of the medial edge epithelial cells was studied in vitro using vital cell labeling and immunohistochemical techniques. Despite clear adherence, the null mutant palatal shelves did not fuse in vitro, but instead the medial edge epithelial cells survived at the midline position, and the basement membrane was resistant towards degradation. Supplementation of the culture medium with the mature form of TGF-beta3 was able to fully correct the defective fusion in the null mutant specimens. Our results demonstrate that the reason for the defective palatal fusion in TGF-beta3 (-/-) samples is not impaired adhesion. Our data define a specific role for TGF-beta3 in the events that control transdifferentiation of the medial edge epithelial cells including degradation of the underlying basement membrane. PMID- 9215641 TI - Spatial and temporal expression of the 72-kDa type IV collagenase (MMP-2) correlates with development and differentiation of valves in the embryonic avian heart. AB - Extracellular proteases may play an important role in the regulation of cell migration and remodeling of the extracellular matrix during development. In this study, we have examined the embryonic avian heart for the expression of matrix metalloproteases. The 72-kDa type IV collagenase, MMP-2, was detected in extracts of whole hearts and showed a modest increase in amount over time. This increase in enzyme activity corresponded to a small increase in the steady-state level of mRNA for this enzyme. A more dramatic increase was seen in the amount of the 66 kDa activated form of this enzyme as development progressed, suggesting that the process of activation, rather than enzyme synthesis, may be the important regulatory step in this system. Coincident with the change in the level of active MMP-2 was a significant increase in the expression of the MMP-2 activator, MT MMP, between stages 12 and 24. The message for MMP-2 was expressed by the endocardium of the cushion tissues which was undergoing an epithelial-mesenchymal transition, and by the migrating mesenchymal cells, suggesting a role for this protease in regulating cell motility and matrix invasion. In older staged hearts, the cells of the differentiating valves expressed high levels of MMP-2 which may be important for the final remodeling events in this region. PMID- 9215643 TI - Spinal muscular atrophy gene wobbler of the mouse: evidence from chimeric spinal cord and testis for cell-autonomous function. AB - Human hereditary neurodegenerative diseases are genetically and mechanistically very heterogeneous and so are spinal muscular atrophies and cerebellar ataxias in the mouse, despite the common phenomenon of neuronal death. In this species, a number of mutations impair spermiogenesis in addition to neuron survival. Among these, the wobbler mutation on proximal chromosome 11 of the mouse leads to motoneuron degeneration in brain stem and spinal cord and to a defect of spermiogenesis. Chimeric mice of the type wr?/wr? <--> +/+ were produced, and their allelic status at the wr locus was determined by PCR diagnosis of a closely linked marker. Two overt chimeras, one female (XX <--> XX) and one male (XY <--> XY) were identified as wr/wr <--> +/+ and analyzed with respect to their pathological phenotype. Although there was patchy astrogliosis in the spinal cords of both chimeras, their motor performances were overtly normal and muscles were without signs of denervation. The male's testes revealed a mosaic pattern of normal and pathological spermatids. As no progeny was derived from wr spermatids, the spermatocytes appear as a primary target of the wr mutation in testis. Our results argue against a humoral mechanism of the wobbler disease and indicate a cell-autonomous action of the wr gene both in testis and in spinal cord. PMID- 9215642 TI - Cloning and expression analysis of cadherin-10 in the CNS of the chicken embryo. AB - A full-length cDNA of a novel cadherin of chicken (cad10) was cloned. The deduced amino acid sequence of the putative cytoplasmic domain of this molecule is highly homologous to a previously published cytoplasmic fragment of human cadherin-10, a type II cadherin. An in situ hybridization analysis in chicken embryos shows that cad10 expression starts at about 4 days' incubation (E4) and persists at least until the hatching stage. In the central nervous system (CNS), cad10 expression is spatially restricted at all stages of development. At early stages, expression reflects the neuromeric organization of the brain. For example, in the alar plate of the diencephalon, cad10 expression is restricted to the dorsal thalamic neuromere. A number of cad10-expressing brain nuclei are formed in this neuromeric domain during later development. Specific cad10-expressing gray matter structures are also found in all other major divisions of the brain. Many of these structures are known to be functionally connected to each other. The cad10 expression pattern is distinct from that of other cadherins. These results support the idea that cadherins provide a molecular code for the regionalization of the embryonic CNS at the different stages of development. PMID- 9215644 TI - Expression of smooth muscle alpha-actin in mesenchymal cells during formation of avian endocardial cushion tissue: a role for transforming growth factor beta3. AB - During early cardiac morphogenesis, outflow tract (OT) and atrio-ventricular (AV) endothelial cells differentiate into mesenchymal cells, which have characteristics of smooth muscle-like myofibroblasts, and which form endocardial cushion tissue, the primordia of valves, and septa in the adult heart. During this embryonic event, transforming growth factor beta3 (TGF beta3) is an essential element in the progression of endothelial-transformation into mesenchyme. TGF beta(s) are known to be a potent inducer for mesodermal differentiation and a promoter for differentiation of endothelial cells into smooth muscle-like cells. Using a monoclonal antibody against smooth muscle specific alpha-actin (SMA), we examined the immunohistochemical staining of this form of actin in avian endocardial cushion tissue formation. To determine whether TGF beta3 initiates the expression of SMA, the pre-migratory AV endothelial monolayer was cultured with or without chicken recombinant TGF beta3 and the expression of SMA was examined immunochemically. Migrating mesenchymal cells expressed SMA beneath the cell surface membrane. These cells showed a reduction of endothelial specific marker antigen, QH1. Stationary endothelial cells did not express SMA. The deposition of SMA in the mesenchymal tissue persisted until the end of the fetal period. Pre-migratory endothelial cells cultured in complete medium (CM199) that contained TGF beta3 expressed SMA, whereas cells cultured in CM199 alone did not. At the onset of the endothelial-mesenchymal transformation, migrating mesenchymal cells express SMA and the expression of this form of actin is upregulated by TGF beta3. The induction of the expression of SMA by TGF beta3 is one of the initial events in the cytoskeletal reorganization in endothelial cells which separate from one another during the initial phenotypic change associated with the endothelial-mesenchymal transformation. PMID- 9215645 TI - GAL4 enhancer traps expressed in the embryo, larval brain, imaginal discs, and ovary of Drosophila. AB - We have screened a collection of approximately 400 GAL4 enhancer trap lines for useful patterns of expression in the embryo, larval brain, imaginal discs, and ovary using a UAS-lacZ reporter construct. Although similar patterns of expression have previously been reported in the original P[lacZ] enhancer trap screens, these lines are useful for directing ectopic expression of genes in discrete patterns during these stages. In addition, we have identified some unique patterns of expression that have not been previously reported. PMID- 9215646 TI - Dynamic expression of chicken Sox2 and Sox3 genes in ectoderm induced to form neural tissue. AB - The chick genes, cSox2 and cSox3, are members of a large family of genes that encode transcription factors. Previous studies have shown that these genes are predominantly expressed in the central nervous system during embryonic development. We show that cSox3 is expressed throughout the ectoderm that is competent to form nervous tissue before neural induction. The expression of cSox3 is lost from cells as they undergo gastrulation to form nonectodermal tissues; the transcription factor, Brachyury, appears in cells about to undergo gastrulation a short time before cSox3 transcripts are lost. Therefore, Brachyury expression may act functionally upstream of cSox3 downregulation. cSox3 expression is also lost from non-neuronal ectoderm shortly after the neural plate becomes morphologically apparent. cSox2 expression increases dramatically in the central nervous system as neural ectoderm is established. The appearance of cSox2 in neural ectoderm represents one of the earliest molecular responses to neural induction documented thus far. PMID- 9215647 TI - SIV/HIV-1 hybrid virus expressing the reverse transcriptase gene of HIV-1 remains sensitive to HIV-1-specific reverse transcriptase inhibitors after passage in rhesus macaques. AB - We have previously described an animal model for the therapy of human immunodeficiency virus type 1 (HIV-1) infection with HIV-1-specific reverse transcriptase (RT) inhibitors based on a simian immunodeficiency virus (SIV), in which the RT gene of SIV was replaced by the RT gene of HIV-1. In vitro, replication of the hybrid virus, RT-SHIV, was delayed compared with parental SIV. RT-SHIV could induce AIDS-like symptoms and pathologic alterations in rhesus macaques. Characterization of re-isolates recovered from RT-SHIV-infected macaques one-half year after infection revealed that the re-isolates replicated with kinetics similar to those of SIV. Inefficient processing of the Gag-Pol precursor of RT-SHIV may be one reason for the retarded growth of RT-SHIV, because the protease cleavage site between the protease gene and the RT gene was frequently mutated in the RT-SHIV re-isolates. Adaptation of RT-SHIV to the growth in macaques did not result in a relevant loss of sensitivity to nonnucleoside RT inhibitors (NNRTIs). However, because a minor sub-population of the RT-SHIV re-isolates contained a mutation conferring low-level resistance to ddI and ddC, the RT-SHIV/macaque model may underestimate the efficacy of these drugs. Nevertheless, this report further supports the suitability, reliability, and usefulness of the RT-SHIV/macaque model to investigate the antiviral properties of most RT inhibitors in an in vivo setting. PMID- 9215648 TI - Cytomegalovirus and simian immunodeficiency virus coinfection: longitudinal study of antibody responses and disease progression. AB - Antibody titers to rhesus cytomegalovirus (RhCMV) were prospectively analyzed over a period of 68 weeks in a longitudinal serosurvey of 17 RhCMV-seropositive rhesus macaques (Macaca mulatta) experimentally coinfected with simian immunodeficiency virus (SIV). These were compared with anti-RhCMV titers in 18 animals that were also naturally infected with RhCMV but not infected with SIV. Fluctuations in anti-RhCMV antibody titers were observed within 5 weeks of SIV inoculation, and two distinct patterns of RhCMV antibody response were observed in SIV-infected animals. Animals showing a progressive decline in anti-RhCMV immunoglobulin G (IgG) exhibited the most rapid disease progression, coincident with low anti-SIV and anti-tetanus toxoid IgG responses, high levels of p27 antigen in the plasma, and short survival. Animals exhibiting a more stable CMV specific response after SIV inoculation had the least rapid disease course. Anti RhCMV antibody titers in SIV-uninfected animals remained relatively stable during the period of study. Evidence that preinoculation immunologic measures predicted postinoculation outcome was equivocal. PMID- 9215649 TI - Inhibition of HIV infection of H9 cells by chlorpromazine derivatives. AB - The binding between the HIV surface protein, gp120, and the CD4 coreceptor is known to be initiated by electrostatic interactions. Because of the ability of chlorpromazine to interact with proteins by charge transfer, we tested several derivatives for their ability to block binding of HIV to CD4+ cells. We have shown that 7,8-dioxo-chlorpromazine blocks binding of fluorescein isothiocyanate labeled anti-Leu3a and rgp120 to peripheral human blood T4 cells and blocks syncytia formation between gp120- and CD4-expressing cells. We also found that 7,8-dioxo-chlorpromazine blocks HIV infectivity of H9 cells and acts synergistically with zidovudine. PMID- 9215650 TI - 1H magnetic resonance spectroscopy reveals neuronal injury in a simian immunodeficiency virus macaque model. AB - Infection with human immunodeficiency virus (HIV) commonly results in neurologic disease called the AIDS dementia complex. Neuronal loss and injury have been found in the HIV brain, but the underlying mechanisms are not understood. The simian immunodeficiency virus (SIV)-infected macaque is an excellent animal model for HIV infection, but neuronal loss has not been demonstrated. To determine whether neuronal damage occurs in the SIV brain, we quantified the neuronal marker N-acetylaspartate (NAA) using proton magnetic resonance spectroscopy (1H MRS) in brain extracts of control and SIV-infected macaques and correlated these findings with histologic analyses. We found reduced NAA in the SIV-infected animals compared with controls (2.94 +/- 1.37 versus 6.21 +/- 1.73 micromol/g of wet weight; p = 0.004). A significant decrease in NAA was also found in SIV infected animals sacrificed in the acute stages of infection 9 or 10 days after inoculation with SIVmacYnef. We conclude that SIV infection of rhesus macaques results in neuronal damage that is demonstrable shortly after infection and that 1H-MRS may be used to measure such injury. The results further support the SIV macaque as a useful model to study the mechanisms of neuropathogenesis by HIV. PMID- 9215651 TI - Phase I/II study of the toxicity, pharmacokinetics, and activity of the HIV protease inhibitor SC-52151. AB - SC-52151, an HIV-1 protease inhibitor, was developed as an ethanol-based elixir and subsequently as a self-emulsifying drug delivery system (SEDDS) to improve bioavailability. To evaluate formulation and treatment regimen effects, we conducted a four-arm, phase I/II study using the highest previously tested daily dose, 2250 mg. Forty-nine patients received the elixir or SEDDS at a dosage of 750 mg three times daily or 1125 mg twice daily for 14 days. One patient developed hypertriglyceridemia, and one had fever and dyspnea. The SEDDS formulation compared with the elixir resulted in a larger area under the concentration-time curve (AUC, p < 0.001), peak (Cmax, p = 0.041) and trough (Cmin, p = 0.025). Twice-daily administration compared with administration three times daily produced a higher cumulative AUC (p = 0.008). Both SEDDS regimens produced mean plasma concentrations above the 90% inhibitory concentration (IC90) for HIV. A mean decline of 0.03 log10 RNA copies (SEDDS) and an increase of 0.15 log10 (elixir) were observed. Although SC-52151 was well tolerated and the SEDDS formulation resulted in plasma concentrations above the IC90 for viral replication, no antiviral activity was produced. PMID- 9215652 TI - Transmission of HIV-1 in infants born to seropositive mothers: PCR-amplified proviral DNA detected by flow cytometric analysis of immunoreactive beads. AB - The diagnosis of HIV infection in newborns is established by amplification of proviral DNA using the polymerase chain reaction (PCR). We developed a nonisotopic method for heminested PCR using a biotinylated primer among sets of three oligonucleotides, each selected from the HIV long terminal repeat (LTR) and gag sequences. An internal probe incorporating digoxigenin-dUTP was also synthesized by PCR. The PCR products, hybridized with LTR region or gag region probes, were captured with streptavidin-coated magnetic beads and detected by fluorescein isothiocyanate-labeled antidigoxigenin in flow cytometric analysis. This immunoreactive bead assay (PCR-IRB) detected about three copies of HIV proviral DNA. A panel of 50 coded DNA specimens of infants previously assayed by conventional PCR and with known clinical results revealed that the PCR-IRB findings using LTR, but not gag, were in agreement. A double-blind prospective study of blood samples from 14 mother-infant pairs using the PCR-IRB amplification of LTR gave results similar to the commercial Amplicor HIV-1 PCR test and were consistent with the clinical outcomes. PCR-IRB results were positive for 11 mothers and three infants, one at birth, one at 2 weeks after birth, and one at 8 weeks after birth. PCR-IRB is a simple, reliable, specific, and automatable assay useful in the early diagnosis of perinatal HIV infection in clinical practice and regional screening programs. PMID- 9215653 TI - Cross-sectional and longitudinal evaluation of body composition in men with HIV infection. AB - Body wasting is an increasingly prevalent AIDS-defining condition and an independent risk factor for mortality in patients infected with HIV. Largely on the basis of studies conducted early in the epidemic, HIV-associated wasting has been assumed to feature a disproportionate loss of lean tissue. We report the results obtained from cross-sectional and longitudinal studies that differ from these earlier observations. In a cross-sectional analysis, weight and body composition determined by dual-energy x-ray absorptiometry and bioelectrical impedance analysis in 32 HIV-infected men with documented weight loss of > or = 10% were compared to those in 46 HIV-positive men without significant weight loss and 32 HIV-negative controls. Fat, lean body mass (LBM), and body cell mass (BCM) were significantly lower in men with weight loss relative to controls (p < 0.001 for fat and BCM; p = 0.01 for LBM). Two thirds of the difference in weight was fat. For the longitudinal analysis, the composition of weight lost over time was evaluated in paired measurements in men grouped by body fat content (<15% or >15%, n = 10 per group). Weight loss in patients with baseline fat of more than 15% was only 16% LBM, but the composition of weight lost in men with baseline fat of less than 15% was 70% LBM. We conclude that progressive decreases in fat and lean tissue occur in men with HIV infection, with the composition of weight lost depending on baseline fat content. These results argue against the widely held notion that HIV-associated wasting is characterized by preservation of fat at the expense of lean tissue. PMID- 9215654 TI - Comparison of serum and plasma viral RNA measurements in primary and chronic human immunodeficiency virus type 1 infection. AB - We sought to define the relation between serum and plasma HIV-1 viral RNA load in patients with primary and chronic HIV-1 disease. HIV-1 viral load was determined from 116 serum and plasma samples, including 33 matched pairs, from five patients with primary and three patients with chronic HIV disease using the Roche HIV Monitor assay. The mean +/- standard deviations of the serum and plasma viral RNA levels from the 33 matched pairs were 4.372 +/- 0.885 and 4.478 +/- 0.950 log10 (copies/ml), respectively. This -0.106 log difference between serum and plasma viral RNA levels, which equates to 21% of non-log-transformed values, was not statistically significant by the Wilcoxon sign rank test (p = 0.09). The distributions of serum and plasma viral load slopes, calculated from all available viral RNA load data for each patient, were also not statistically different (p = 0.07). The levels of HIV-1 RNA measured in the serum or plasma of HIV-seropositive patients yield equivalent biologic information. PMID- 9215655 TI - Effects of reverse transcriptase inhibitor therapy on the HIV-1 viral burden in semen. AB - HIV-1 infection continues to spread worldwide, primarily through sexual intercourse. Because semen is a major vehicle for transmission of HIV-1, we evaluated the effects of reverse transcriptase inhibitor therapy on the amount of HIV-1 in semen. The semen and blood of 11 HIV-1-infected men (i.e. treatment group) were collected before the initiation of reverse transcriptase inhibitor therapy and then 8 to 18 weeks after initiation of therapy. The semen and blood of another 11 HIV-1-infected men (i.e., longitudinal group), who were not on or had no change in antiretroviral therapy for at least 2 months before study entry, were collected at approximately 2-week intervals for 10 to 26 weeks. In the treatment group, 82% of the seminal plasma HIV-1 RNA levels decreased from baseline after 8 to 18 weeks of therapy (median reduction of 1.01 log10, p = 0.01), and 100% of the blood plasma RNA levels decreased from baseline over the same period (median reduction of 0.92 log10, p = 0.003). Five of these patients were followed for at least 52 weeks and had a median seminal plasma HIV-1 RNA level of 0.66 log10 below baseline at 1 year. All subjects in the treatment group with positive cultures at baseline (50%) had negative cultures or a lower infectious units per ejaculate at the 8- to 18-week follow-up examinations. The HIV-1 RNA levels in blood and semen of the longitudinal group did not change significantly over 10 to 26 weeks. Initiation of reverse transcriptase inhibitor therapy effectively reduces shedding of HIV-1 in semen and may therefore reduce the spread of infection within populations. PMID- 9215656 TI - T cells from individuals in advanced stages of HIV-1 infection do not proliferate but express activation antigens in response to HIV-1-specific antigens. AB - Like T cells from healthy subjects, those of HIV-1-infected patients are capable of expressing activation antigens on their surface after antigenic or mitogenic stimulation, but their proliferative activity is strongly reduced or even absent, especially in patients with advanced stages of the disease. The characteristic of expressing activation antigens in response to different stimuli in the absence of cell proliferation is shared by CD4+ and CD8+ T-cell subsets from HIV-1-infected patients. The number of T cells capable of expressing CD25 and CD71 in response to HIV-1-related antigens but not of proliferating increased significantly with the progression of the disease, but the number of T cells capable of expressing the two activation antigens in response to the classic tetanus toxoid recall antigen decreased. The higher numbers of T cells capable of responding to HIV-1 related antigens in conjunction with a reduction in the number of T cells responding to recall antigens may explain the occurrence of different infections, including opportunistic microorganisms, during the more advanced stages of HIV-1 infection. Because the increase in the number of HIV-1 antigen-responding T cells (defined by CD25 and CD71 activation antigen expression) is a characteristic of symptomatic HIV-1-infected patients, expression (by flow cytometry) of these activation antigens on T cells in response to HIV-1 antigens could be used as a new marker of disease progression. PMID- 9215657 TI - Increased expression of interleukin-2 receptor alpha on peripheral blood mononuclear cells in HTLV-I tax/rex mRNA-positive asymptomatic carriers. AB - Using the double-nested reverse transcription-polymerase chain reaction, we assayed human T-cell leukemia virus type I (HTLV-I) tax/rex-encoded mRNA in the peripheral blood mononuclear cells (PBMCs) of asymptomatic carriers as an index of the expression of HTLV-I in vivo in relation to the proviral DNA level. HTLV-I tax/rex mRNA was detected in only 1 (3.3%) of 30 samples with medium or lower proviral DNA levels, but it was detected in 11 (39.3%) of 28 samples with high HTLV-I proviral DNA levels, estimated as equal to or more than the proviral DNA of 10 ng of HUT102 (i.e., HUT102 cells were used as positive controls). The mean number of interleukin-2 receptor alpha (IL-2R alpha)-positive cells as a percentage of the total number of PBMCs was higher (13.2%) in the tax/rex mRNA positive carriers with high proviral DNA levels than in the carriers who were mRNA negative (8.4%) (p = 0.004, Wilcoxon test). These results suggest that virus activation as indicated by the presence of tax/rex mRNA in asymptomatic carriers with high proviral DNA levels is associated with an elevation of the IL-2R alpha positive cells in vivo. PMID- 9215658 TI - Sexual behavior and injection drug use during pregnancy and vertical transmission of HIV-1. AB - We evaluated maternal sexual behavior and injection drug use practices as possible risk factors for vertical transmission of human immunodeficiency virus type 1 (HIV-1). Data were analyzed from the Mothers and Infants Cohort Study, a prospective study in Brooklyn and the Bronx, New York. A total of 207 mother infant sets were enrolled between 1986 and 1991 and followed for up to 4 years after the enrollment visit during pregnancy. HIV-1 transmission occurred in 49 of 201 mother-infant sets, yielding an overall transmission rate of 24.4% (95% confidence interval (CI) = 18.7% to 31.0%). Increased frequency of vaginal intercourse after the first trimester of pregnancy was positively associated with vertical transmission of HIV-1 (trend p = 0.03). A lifetime history of injection drug use was not associated with vertical transmission. However, a history of combined cocaine and heroin injection after the first trimester of pregnancy was associated with vertical HIV-1 transmission, particularly among women with CD4+ lymphocyte levels of 20% or higher (risk ratio = 4.0; 95% CI = 2.0 to 8.1). Cocaine and heroin injection drug use after the first trimester accounted for most of the relation between preterm birth and vertical HIV-1 transmission in this cohort. Maternal coinfection with hepatitis C virus or human T-cell lymphotropic virus types I and II could not explain these observations, because coinfection with these viruses had no detectable effect on HIV-1 transmission. These results suggest that maternal sexual behavior and injection drug use practices during the second and third trimester of pregnancy may modify the risk of vertical HIV-1 transmission. PMID- 9215659 TI - Antisulfatide IgG antibodies recognize HIV proteins. PMID- 9215660 TI - Successful use of cidofovir in treating AIDS-related cytomegalovirus retinitis, encephalitis, and esophagitis. PMID- 9215662 TI - Birth weight adjustments for gestational age. The European Collaborative Study. PMID- 9215661 TI - Effects of vitamin A supplementation on viral load in HIV-1-infected pregnant women. PMID- 9215663 TI - Prevalence of human T-cell lymphotropic virus type I infection among volunteer blood donors in Kuwait. PMID- 9215664 TI - Transition from premutation to full mutation in fragile X syndrome is likely to be prezygotic. AB - In the fragile X syndrome, the transition from unmethylated moderate expansions of the CGG repeat (premutations) to methylated large expansions (full mutations) occurs only through maternal transmission. The risk of such transition is highly correlated with the size of the maternal premutation (PM), being very low for small PM alleles (approximately 60 repeats), to 100% for alleles above 100 repeats. The timing of this transition was the object of much speculation. A postzygotic transition was proposed as a preferred model, based on the observation that males with full mutation (FM) have PM in sperm. Analysis of tissues from affected fetuses, including additional data reported here, indicate that such a putative postzygotic transition would have to occur very early in embryogenesis and most likely before determination of germ cell lineage. At least 15% of carriers of a FM show a significant proportion of white blood cells carrying a PM (mutation mosaics). We performed a simulation study showing that, if transition to FM is postzygotic, one should observe a much higher proportion of such mosaics in offspring of mothers with small PMs. This was compared with the actual pattern observed in 212 mutated offspring of 112 PM carrier mothers. We found no effect of maternal PM size on incidence of mosaicism in leucocytes. We propose that this is strong, albeit indirect evidence against a postzygotic transition to FM. A transition at an early morula stage (before day 3) cannot, however, be formally excluded. PMID- 9215665 TI - Visual pigment gene structure and expression in human retinae. AB - We determined the genotypes of the X-chromosome-linked red/green color vision genes by a novel PCR/SSCP-based method and assessed expression by mRNA analysis in retinae of 51 unselected post mortem eye specimens from Caucasian males of unknown color vision status. All individuals had a single red (long-wave) pigment gene and one or more (an average of two) green (middle-wave) pigment genes. Four males had 5'green-red3' hybrid genes in addition to normal red and green pigment genes. These findings are consistent with earlier studies on human visual pigment gene structure using Southern blotting and with a recent study using pulsed-field electrophoresis. We interpret claims of much larger numbers of red, green and green-red hybrid genes to be technical artifacts. The ratio of expressed red to green pigment retinal mRNA varied widely (1-10 with a mode of 4) and was not correlated with that of red to green pigment genes. In one individual with a green-red hybrid gene in addition to normal red and green pigment genes, the normal red pigment gene and the hybrid gene were both expressed, but the normal green gene was not. This person presumably had deuteranomalous color vision. Two with green-red hybrid genes expressed the normal red and green pigment genes, but not the hybrid genes. These two individuals presumably had normal color vision. We interpret the failure to express their green-red hybrid genes to be caused by their location at a more distal position in the visual pigment gene array. PMID- 9215666 TI - Interchromosomal duplications of the adrenoleukodystrophy locus: a phenomenon of pericentromeric plasticity. AB - A 9.7 kb segment encompassing exons 7-10 of the adrenoleukodystrophy (ALD) locus of the X chromosome has duplicated to specific locations near the pericentromeric regions of human chromosomes 2p11,10p11, 16p11 and 22q11. Comparative sequence analysis reveals 92-96% nucleotide identity, indicating that the autosomal ALD paralogs arose relatively recently during the course of higher primate evolution (5-10 million years ago). Analysis of sequences flanking the duplication region identifies the presence of an unusual GCTTTTTGC repeat which may be a sequence specific integration site for the process of pericentromeric-directed transposition. The breakpoint sequence and phylogenetic analysis predict a two step transposition model, in which a duplication from Xq28 to pericentromeric 2p11 occurred once, followed by a rapid distribution of a larger duplicon cassette among the pericentromeric regions. In addition to facilitating more effective mutation detection among ALD patients, these findings provide further insight into the molecular basis underlying a pericentromeric-directed mechanism for non-homologous interchromosomal exchange. PMID- 9215667 TI - Evidence by allelic association-dependent methods for a type 1 diabetes polygene (IDDM6) on chromosome 18q21. AB - Type 1 diabetes is a common polygenic disease. Fine mapping of polygenes by affected sibpair linkage analysis is not practical and allelic association or linkage disequilibrium mapping will have to be employed to attempt to detect founder chromosomes. Given prior evidence of linkage of the Jk-D18S64 region of chromosome 18q12-q21 to type 1 diabetes, we evaluated the 12 informative microsatellite markers in the region for linkage with disease by the transmission disequilibrium test (TDT) in a UK data set of type 1 diabetic families (n = 195). Increased transmission of allele 4 of marker D18S487 to affected children was detected (P = 0.02). Support for this was extended in a total of 1067 families from four different countries by isolating, and evaluating by the TDT, two novel microsatellites within 70 kb of D18S487. Evidence for linkage and association was P = 5 x 10(-5) and 3 x 10(-4), respectively. There was no evidence for increased transmission of associated alleles to nonaffected siblings. Analysis of an additional 390 families by the TDT did not extend the evidence further, and reduced support in the total 1457 families to P = 0.001 for linkage and P = 0.003 for association. However, evidence for linkage by affected sibpair allele sharing was strong (P = 3.2 x 10(-5)) in the second data set. Heterogeneity in TDT results between data sets was, in part, accounted for by the presence of more than one common disease-associated haplotype (allelic heterogeneity) which confounds the analysis of individual alleles by the TDT. Guidelines for strategies for the mapping of polygenes are suggested with the emphasis on collections of large numbers of families from multiple populations that should be as genetically homogeneous as possible. PMID- 9215668 TI - Evidence for a type 1 diabetes susceptibility locus (IDDM10) on human chromosome 10p11-q11. AB - A region of linkage to type 1 diabetes has been defined on human chromosome 10p11 q11 (IDDM10; P = 0.0007) using 236 UK and 76 US affected sibpairs and a 1 cM resolution microsatellite marker map. Analysis by the transmission disequilibrium test (TDT) in 1159 families with at least one diabetic child, from the UK, the US, Norway, Sardinia and Italy provided additional support for linkage at D10S193 (P = 0.006, Pc = 0.17). Notably, 5.1 cM distal to D10S193, marker D10S588 also provided positive TDT results (P = 0.009, Pc = 0.25) but the allele under analysis was also preferentially transmitted to nonaffected siblings (P = 0.0008, Pc = 0.02). This allele was positively associated in an independent UK case control study and, importantly, was neutrally transmitted in control CEPH families. These results suggest a type 1 diabetes susceptibility locus on chromosome 10p11-q11 (provisionally designated IDDM10) and demonstrate the necessity of analysis of non affected siblings in disease families, as well as analysis of control families. PMID- 9215669 TI - The molecular basis of the Kidd blood group polymorphism and its lack of association with type 1 diabetes susceptibility. AB - The Kidd blood group locus encodes a urea transporter which is expressed on human red cells and in the kidney. This gene is located on chromosome 18q12, and evidence for linkage and association with type 1 diabetes mellitus has been reported. To investigate this further, the genetic basis for the blood group Jk(a)/Jk(b) polymorphism was first determined by sequencing reverse-transcribed reticulocyte RNAs from Jk(a+b-) and Jk(a-b+) donors. The Jk(a)/Jk(b) polymorphism was caused by a transition (G838A), resulting in a Asp280Asn amino acid substitution and an MnlI restriction fragment length polymorphism (RFLP). Using the MnlI RFLP, we found that the Jk(a)/Jk(b) polymorphism was not in linkage disequilibrium with type 1 diabetes in 228 multiplex UK and US families tested. PMID- 9215670 TI - Elastin point mutations cause an obstructive vascular disease, supravalvular aortic stenosis. AB - Supravalvular aortic stenosis (SVAS) is an inherited obstructive vascular disease that affects the aorta, carotid, coronary and pulmonary arteries. Previous molecular genetic data have led to the hypothesis that SVAS results from mutations in the elastin gene, ELN. In these studies, the disease phenotype was linked to gross DNA rearrangements (35 and 85 kb deletions and a translocation) in three SVAS families. However, gross rearrangements of ELN have not been identified in most cases of autosomal dominant SVAS. To define the spectrum of ELN mutations responsible for this disorder, we refined the genomic structure of human ELN and used this information in mutational analyses. ELN point mutations co-segregate with the disease in four familial cases and are associated with SVAS in three sporadic cases. Two of the mutations are nonsense, one is a single base pair deletion and four are splice site mutations. In one sporadic case, the mutation arose de novo. These data demonstrate that point mutations of ELN cause autosomal dominant SVAS. PMID- 9215671 TI - Elastin: genomic structure and point mutations in patients with supravalvular aortic stenosis. AB - We describe the complete exon-intron structure of the human elastin (ELN) gene located at chromosome 7q11.23. There are 34 exons occupying approximately 47 kb of genomic DNA. All exons are in-frame, allowing exon skipping without disrupting the reading frame. Microsatellites are located in introns 17 and 18. Deletions of all or large parts of the ELN gene have been previously reported in two patients with supravalvular aortic stenosis (SVAS), and SVAS is also a frequent feature of Williams syndrome, where patients are hemizygous for ELN. We list primer pairs for amplifying each exon, with flanking intron, from genomic DNA to allow detection of point mutations in the ELN gene. We show that some patients with isolated SVAS have point mutations that are predicted to lead to premature chain termination. Knowledge of the genomic structure will allow more extensive mutation screening in genomic DNA of patients with SVAS and other conditions. PMID- 9215672 TI - Molecular basis of the brindled mouse mutant (Mo(br)): a murine model of Menkes disease. AB - The brindled mouse mutant (Mo(br)) is the closest animal model of the human genetic copper deficiency, Menkes disease, which is presumed to be due to a mutation at the X-linked mottled locus (Mo). The mutant mice are hypopigmented and die at around 15 days after birth, but can be saved by treatment with copper before the 10th postnatal day. Menkes disease has been shown to be due to mutations of the gene ATP7A which encodes P-type ATPase (referred to here as MNK). MNK is likely to function in copper efflux from cells, but the full range of its biological activity is not fully understood. The nature of the mutation in the brindled mouse is of importance in our understanding of the role of MNK and for devising treatment strategies for Menkes disease. Here we show that the brindled mouse has a deletion of two amino acids in a highly conserved, but functionally uncharacterized, region of Mnk. Comparison with the Ca ATPases suggests this region may be involved in conformational changes associated with the E1/E2 transition fundamental to the action of P-type ATPases. We also describe the first Western blot data for Mnk in tissues, and these show normal levels of Mnk in mutant and brindled kidneys but none in liver. In the kidney, immunohistochemistry demonstrated Mnk in the proximal and distal tubules, the distribution is identical in mutant and normal. This distribution is consistent with Mnk being involved in copper resorption from the urine. PMID- 9215673 TI - Developmental expression of the mouse mottled and toxic milk genes suggests distinct functions for the Menkes and Wilson disease copper transporters. AB - Menkes disease and Wilson disease are human disorders of copper transport caused by mutations in distinct genes encoding similar copper-transporting P-type ATPases. These genes are expressed in different adult tissues in patterns reflecting disease manifestations. The mouse homologues for the Menkes (MNK) and Wilson (WND) disease genes are the mottled (Atp7a) and toxic milk (Atp7b) genes, respectively. Using RNA in situ hybridization we describe the distribution of mottled and toxic milk transcripts during mouse embryonic development. The mottled gene is expressed in all tissues throughout embryogenesis and is particularly strong in the choroid plexuses of the brain. Mottled expression in the liver is in contrast to the prior observation of absent or very low expression in the adult liver. Expression of the toxic milk gene is significantly more delimited, with early expression in the central nervous system, heart and liver. Later in gestation, toxic milk transcript is clearly seen in the liver, intestine, thymus and respiratory epithelium including nasopharynx, trachea and bronchi. In lung, toxic milk expression is restricted to bronchi, while mottled expression is diffuse. Hepatic expression of both toxic milk and mottled is in the parenchyma, as opposed to blood cells. These results suggest that the mottled gene product functions primarily in the homeostatic maintenance of cell copper levels, while the toxic milk gene product may be specifically involved in the biosynthesis of distinct cuproproteins in different tissues. PMID- 9215674 TI - Genetic predisposition to phaeochromocytoma: analysis of candidate genes GDNF, RET and VHL. AB - Inherited predisposition to phaeochromocytoma (MIM No 171300) occurs in multiple endocrine neoplasia type 2 (MEN 2) (MIM No 171400), von Hippel-Lindau (VHL) disease (MIM No 199300), and neurofibromatosis type 1 (NF1) (MIM No 162200). In addition, familial phaeochromocytoma alone has also been reported and we and others have identified germline VHL mutations in five of six kindreds analysed previously. Germline mutations in the RET proto-oncogene, which encodes a receptor tyrosine kinase, and in the VHL tumour suppressor gene cause MEN 2 and VHL disease, respectively. To further investigate the genetics of phaeochromocytoma predisposition, we analysed three groups of patients with no evidence of VHL disease, MEN 2 or NF1: Group A, eight kindreds with familial phaeochromocytoma; Group B, two patients with isolated bilateral phaeochromocytoma; and Group C, six cases of multiple extra-adrenal phaeochromocytoma or adrenal phaeochromocytoma with a family history of neuroectodermal tumours. Germline missense VHL mutations were identified in three of eight kindreds with familial phaeochromocytoma. A germline VHL mutation was also characterised in one of the two patients with bilateral phaeochromocytoma. No VHL or RET mutations were detected in the final group of patients with multiple extra-adrenal phaeochromocytoma or adrenal phaeochromocytoma with a family history of neuroectodermal tumours. The absence of germline VHL and RET gene mutations in many of these families suggested that other phaeochromoeytoma susceptibility loci may exist. Glial cell line-derived neurotrophic factor (GDNF) has been recently identified as a natural ligand for RET. Thus, it seems plausible that GDNF is a good candidate gene to play a role in phaeochromocytoma susceptibility. We searched for germline mutations in GDNF in 16 cases of familial phaeochromocytoma (groups A, B and C) and looked for evidence of somatic change in GDNF in 28 sporadic phaeochromocytomas, 12 MEN 2 phaeochromocytomas and five VHL phaeochromocytomas. No GDNF mutations were identified in patients with familial phaeochromocytoma disease, but a c277C-->T (R93W) sequence variant was identified in one of 28 sporadic tumours. This candidate mutation was identified in the germline and tumour tissue but was not present in 104 control GDNF alleles. GDNF sequence variants including R93W have been suggested previously to represent low penetrance susceptibility mutations for Hirschsprung disease and the R93W was not identified in 376 control alleles studied by others. These findings suggest that although GDNF mutations do not appear to have a major role in the pathogenesis of familial or sporadic phaeochromocytomas, allelic variation at the GDNF locus may modify phaeochromocytoma susceptibility. PMID- 9215675 TI - Isolation and characterisation of the NBR2 gene which lies head to head with the human BRCA1 gene. AB - To study the regulation of BRCA1 gene expression and the potential importance of dysregulation of this gene in breast and ovarian cancer, we have examined the 5' region of the human BRCA1 gene in detail. We have identified a new gene, NBR2, which is partially related to the NBR1 gene (formerly known as 1A1-3B and mapping directly adjacent to the pseudo-BRCA1 gene) and which lies head to head with the BRCA1 gene. The physical distance between the transcription start sites of the NBR2 and BRCA1 genes is 218 bp, suggesting that regulation of the expression of both genes may be co-ordinated through a bi-directional promoter. The NBR2 gene contains five exons spanning a genomic region of approximately 30 kb between the BRCA1 and pseudo-BRCA1 genes. Northern analysis showed that the NBR2 gene is expressed in all the tissues examined. The NBR2 cDNA contains an open reading frame of 112 amino acids and is predicted to encode a protein of approximately 12 kDa. Single-strand conformation polymorphism (SSCP) analysis of the NBR2 gene failed to identify any mutations in either breast or ovarian cancer, suggesting that if the NBR2 gene is involved in the development of these cancers, other mechanisms for tumorigenesis may exist. Hybridisation of NBR2 probes to zoo blots showed that the NBR2 gene is present in human and other primates. No hybridisation to DNA from other species was observed, suggesting that genomic elements controlling BRCA1 expression may differ between species. PMID- 9215676 TI - Single sperm analysis of the CAG repeats in the gene for Machado-Joseph disease (MJD1): evidence for non-Mendelian transmission of the MJD1 gene and for the effect of the intragenic CGG/GGG polymorphism on the intergenerational instability. AB - To investigate the mechanism of the meiotic instability of expanded CAG repeats in the gene for Machado-Joseph disease (MJD1), we analyzed the CAG repeat sizes of 1036 single sperm from six individuals with Machado-Joseph disease (MJD). The segregation ratio between single sperm with an expanded allele and those with a normal allele is significantly different (P <0.0001) from the expected 1:1 segregation ratio, which demonstrates segregation distortion of expanded alleles in male meiosis. In single sperm from individuals with the [expanded (CAG)n CGG]/[normal (CAG)n-GGG] genotype, significantly greater instability of the CAG repeat was observed compared with single sperm from individuals with the [expanded (CAG)n-CGG]/[normal (CAG)n-CGG] genotype (F-test, P <0.001). These findings in single sperm confirm non-Mendelian transmission of the MJD1 gene and the effect of the intragenic CGG/GGG polymorphism on the intergenerational instability of the CAG repeats in the MJD1 gene, which have been observed in clinical and genetic studies. Our results indicate similarities and dissimilarities between MJD and Huntington's disease or myotonic dystrophy in terms of the inter-allelic interaction, segregation distortions and size distribution of trinucleotide repeats in mutant alleles. Further study is required to determine whether there is a common mechanism underlying the instability of the triplet repeats in 'triplet repeat diseases'. PMID- 9215677 TI - SOX22 is a new member of the SOX gene family, mainly expressed in human nervous tissue. AB - SOX (SRY box-containing) genes share a particular DNA-binding domain, called HMG, with the mammalian testis-determining gene SRY Several SOX genes have already been shown to be transcription factors involved in the decision of important cell fates during development. Here we report the cloning of a new human member of the SOX gene family, SOX22. The corresponding protein contains several domains that are also present in other paralogous SOX proteins. The SOX22 gene maps to chromosome 20 on band p13 and does not appear to be clustered with any other SOX gene mapped to date. SOX22 mRNA is expressed in various fetal and adult organs and tissues, suggesting that this gene plays roles in both differentiation and maintenance of several cell types. PMID- 9215678 TI - Translocation breakpoint maps 5 kb 3' from TWIST in a patient affected with Saethre-Chotzen syndrome. AB - Saethre-Chotzen syndrome, a common autosomal dominant craniosynostosis in humans, is characterized by brachydactyly, soft tissue syndactyly and facial dysmorphism including ptosis, facial asymmetry, and prominent ear crura. Previously, we identified a yeast artificial chromosome that encompassed the breakpoint of an apparently balanced t(6;7) (q16.2;p15.3) translocation associated with a mild form of Saethre-Chotzen syndrome. We now describe, at the DNA sequence level, the region on chromosome 7 affected by this translocation event. The rearrangement occurred approximately 5 kb 3' of the human TWIST locus and deleted 518 bp of chromosome 7. The TWIST gene codes for a transcription factor containing a basic helix-loop-helix (b-HLH) motif and has recently been described as a candidate gene for Saethre-Chotzen syndrome, based on the detection of mutations within the coding region. Potential exon sequences flanking the chromosome 7 translocation breakpoint did not hit known genes in database searches. The chromosome rearrangement downstream of TWIST is compatible with the notion that this is a Saethre-Chotzen syndrome gene and implies loss of function of one allele by a positional effect as a possible mechanism of mutation to evoke the syndrome. PMID- 9215679 TI - Mutations in the Chediak-Higashi syndrome gene (CHS1) indicate requirement for the complete 3801 amino acid CHS protein. AB - Chediak-Higashi syndrome (CHS) is a rare, usually fatal, autosomal recessive disorder characterized by severe immunologic defects, reduced pigmentation, progressive neurologic dysfunction and a bleeding diathesis. The hallmark of CHS is giant organelles and giant granules in many different cell types, most likely the result of defective trafficking of specific organellar and granular proteins necessary for the normal genesis, structure or function of these cytoplasmic components. The CHS1 gene has recently been identified and shown to be homologous to the beige locus of the mouse; however, there has been disagreement as to the length of the functional CHS1 mRNA and protein. Here we report homozygous CHS1 gene mutations in two of the original probands we used to map the gene to 1q42 q44. One of these, a frameshift at codon 3197, supports our assertion that the functional CHS protein is a predicted 3801 amino acid polypeptide encoded by a 13.5 kb mRNA. PMID- 9215680 TI - Identification of mutations in two major mRNA isoforms of the Chediak-Higashi syndrome gene in human and mouse. AB - Chediak-Higashi syndrome is an autosomal recessive, immune deficiency disorder of human (CHS) and mouse (beige, bg) that is characterized by abnormal intracellular protein transport to, and from, the lysosome. Recent reports have described the identification of homologous genes that are mutated in human CHS and bg mice. Here we report the sequences of two major mRNA isoforms of the CHS gene in human and mouse. These isoforms differ both in size and in sequence at the 3' end of their coding domains, with the smaller isoform (approximately 5.8 kb) arising from incomplete splicing and reading through an intron. These mRNAs also differ in tissue distribution of transcription and in predicted biological properties. Novel mutations were identified within the region of the coding domain common to both isoforms in three CHS patients: C-->T transitions that generated stop codons (R50X and Q1029X) were found in two patients, and a novel frameshift mutation (deletion of nucleotides 3073 and 3074 of the coding domain) was found in a third. Northern blots of lymphoblastoid mRNA from CHS patients revealed loss of the largest transcript (approximately 13.5 kb) in two of seven CHS patients, while the small mRNA was undiminished in abundance. These results suggest that the small isoform alone cannot complement Chediak-Higashi syndrome. PMID- 9215681 TI - Polymorphisms in the apolipoprotein(a) gene and their relationship to allele size and plasma lipoprotein(a) concentration. AB - Genotypes at five previously described polymorphic sites at the apolipoprotein(a) gene locus have been determined for the members of 27 families as well as for unrelated white Caucasian and Asian-Indian subjects, and their relationship with isoform size and plasma lipoprotein(a) concentrations investigated. There was strong linkage disequilibrium between sites at the 5'-region of the gene and also between this region and a site in the coding sequence for Kringle 4-37 on the other side of the polymorphic Kringle 4 repeat region. There was no evidence that changes at any of the sites had any direct effect upon lipoprotein(a) concentration. However, certain haplotypes were present almost exclusively on apolipoprotein(a) alleles within a restricted range of sizes and associated lipoprotein(a) concentrations. After correcting for the effect of allele size, there were clear differences between the lipoprotein(a) concentrations associated with alleles of different haplotypes, suggesting that there may be genetically distinct groups of apolipoprotein alleles of different size and different levels of expression. Factors that regulate expression apparently exchange at a rate similar to the rate of change of Kringle 4 repeat number. PMID- 9215682 TI - Autosomal glycogenosis of liver and muscle due to phosphorylase kinase deficiency is caused by mutations in the phosphorylase kinase beta subunit (PHKB). AB - Glycogen storage disease due to phosphorylase kinase deficiency occurs in several variants that differ in mode of inheritance and tissue-specificity. This heterogeneity is suspected to be largely due to mutations affecting different subunits and isoforms of phosphorylase kinase. The gene of the ubiquitously expressed beta subunit, PHKB, was a candidate for involvement in autosomally transmitted phosphorylase kinase deficiency of liver and muscle. To identify such mutations, the complete PHKB coding sequence was amplified by RT-PCR of RNA isolated from blood samples of patients and analyzed by direct sequencing of PCR products. The characterization of mutations was complemented by PCR of genomic DNA. In one female and four male patients, we identified five independent nonsense mutations (Y418ter; R428ter; Y974H+E975ter; Q656ter in two cases), one single-base insertion in codon N421, one splice-site mutation affecting exon 31, and a large deletion involving the loss of exon 8. Although these severe translation-disrupting mutations occur in constitutively expressed sequences of the only known beta subunit gene of phosphorylase kinase, PHKB, they are associated with a surprisingly mild clinical phenotype, affecting virtually only the liver, and relatively high residual enzyme activity of approximately 10%. PMID- 9215683 TI - Human MSH2 binds to trinucleotide repeat DNA structures associated with neurodegenerative diseases. AB - The expansion of trinucleotide repeat sequences is associated with several neurodegenerative diseases. The mechanism of this expansion is unknown but may involve slipped-strand structures where adjacent rather than perfect complementary sequences of a trinucleotide repeat become paired. Here, we have studied the interaction of the human mismatch repair protein MSH2 with slipped strand structures formed from a triplet repeat sequence in order to address the possible role of MSH2 in trinucleotide expansion. Genomic clones of the myotonic dystrophy locus containing disease-relevant lengths of (CTG)n x (CAG)n triplet repeats were examined. We have constructed two types of slipped-strand structures by annealing complementary strands of DNA containing: (i) equal numbers of trinucleotide repeats (homoduplex slipped structures or S-DNA) or (ii) different numbers of repeats (heteroduplex slipped intermediates or SI-DNA). SI-DNAs having an excess of either CTG or CAG repeats were structurally distinct and could be separated electrophoretically and studied individually. Using a band-shift assay, the MSH2 was shown to bind to both S-DNA and SI-DNA in a structure-specific manner. The affinity of MSH2 increased with the length of the repeat sequence. Furthermore, MSH2 bound preferentially to looped-out CAG repeat sequences, implicating a strand asymmetry in MSH2 recognition. Our results are consistent with the idea that MSH2 may participate in trinucleotide repeat expansion via its role in repair and/or recombination. PMID- 9215684 TI - Modulation of disease severity of dystrophic epidermolysis bullosa by a splice site mutation in combination with a missense mutation in the COL7A1 gene. AB - Dystrophic epidermolysis bullosa (EBD) is a clinically heterogeneous skin disorder, characterized by abnormal anchoring fibrils (AF) and loss of dermal epidermal adherence. EBD has been linked to the COL7A1 gene at chromosome 3p21 which encodes collagen VII, the major component of the AF. Here we investigated two unrelated EBD families with different clinical phenotypes and novel combinations of recessive and dominant COL7A1 mutations. Both families shared the same recessive heterozygous 14 bp deletion at the exon-intron 115 boundary of the COL7A1 gene. The deletion caused in-frame skipping of exon 115 and the elimination of 29 amino acid residues from the pro-alpha1(VII) polypeptide chain. As a result, procollagen VII was not converted to collagen VII and the C-terminal NC-2 propeptide which is normally removed from the procollagen VII prior to formation of the anchoring fibrils was retained in the skin. All affected individuals also carried missense mutations in exon 73 of COL7A1 which lead to different glycine-to-arginine substitutions in the triple-helical domain of collagen VII. Combination of the deletion mutation with a G2009R substitution resulted in a mild phenotype. In contrast, combination of the deletion with a G2043R substitution led to a severe phenotype. The G2043R substitution was a de novo mutation which alone caused a mild phenotype. Thus, different combinations of dominant and recessive COL7A1 mutations can modulate disease activity of EBD and alter the clinical presentation of the patients. PMID- 9215685 TI - A novel human serine-threonine phosphatase related to the Drosophila retinal degeneration C (rdgC) gene is selectively expressed in sensory neurons of neural crest origin. AB - Through our transcriptional mapping effort in the Xp22 region, we have isolated by exon trapping a new transcript highly homologous to the Drosophila retinal degeneration C (rdgC) gene. rdgC encodes a serine/threonine phosphatase protein and is required in Drosophila to prevent light-induced retinal degeneration. This human gene is the first mammalian member of the serine-threonine phosphatase with EF hand motif gene family, and was thus named PPEF (Protein Phosphatase with EF calcium-binding domain). The expression pattern of the mouse Ppef gene was studied by RNA in situ hybridization on embryonic tissue sections. While rdgC is expressed in the visual system of the fly, as well as in the mushroom bodies of the central brain, we found that Ppef is highly expressed in sensory neurons of the dorsal root ganglia (DRG) and neural crest-derived cranial ganglia. The selective pattern of expression makes PPEF an important marker for sensory neuron differentiation and suggests a role for serine-threonine phosphatases in mammalian development. PMID- 9215686 TI - Missense mutations in the human glutathione synthetase gene result in severe metabolic acidosis, 5-oxoprolinuria, hemolytic anemia and neurological dysfunction. AB - Severe glutathione synthetase (GS) deficiency is a rare genetic disorder with neonatal onset. The enzymatic block of the gamma-glutamyl cycle leads to a generalized glutathione deficiency. Clinically affected patients present with severe metabolic acidosis, 5-oxoprolinuria, increased rate of hemolysis and defective function of the central nervous system. The disorder is inherited in an autosomal recessive mode and, until recently, the molecular basis has remained unknown. We have sequenced 18 GS alleles associated with enzyme deficiency and we detected missense mutations by direct sequencing of cDNAs and genomic DNA. In total, 13 different mutations were identified. Four patients were found to be compound heterozygotes and two individuals were apparently homozygous. Reduced enzymatic activities were demonstrated in recombinant protein expressed from cDNAs in four cases with different missense mutations. The results from biochemical analysis of patient specimens, supported by the properties of the expressed mutant proteins, indicate that a residual activity is present in affected individuals. Our results suggest that complete loss of function of both GS alleles is probably lethal. It is postulated that missense mutations will account for the phenotype in the majority of patients with severe GS deficiency. PMID- 9215687 TI - Incomplete rescue of cystic fibrosis transmembrane conductance regulator deficient mice by the human CFTR cDNA. AB - We have used a mouse model to study the ability of human CFTR to correct the defect in mice deficient of the endogenous protein. In this model, expression of the endogenous Cftr gene was disrupted and replaced with a human CFTR cDNA by a gene targeted 'knock-in' event. Animals homozygous for the gene replacement failed to show neither improved intestinal pathology nor survival when compared to mice completely lacking CFTR. RNA analyses showed that the human CFTR sequence was transcribed from the targeted allele in the respiratory and intestinal epithelial cells. Furthermore, in vivo potential difference measurements showed that basal CFTR chloride channel activity was present in the apical membranes of both nasal and rectal epithelial cells in all homozygous knock-in animals examined. Ussing chamber studies showed, however, that the cAMP-mediated chloride channel function was impaired in the intestinal tract among the majority of homozygous knock-in animals. Hence, failure to correct the intestinal pathology associated with loss of endogenous CFTR was related to inefficient functional expression of the human protein in mice. These results emphasize the need to understand the tissue-specific expression and regulation of CFTR function when animal models are used in gene therapy studies. PMID- 9215688 TI - The oral-facial-digital syndrome type 1 (OFD1), a cause of polycystic kidney disease and associated malformations, maps to Xp22.2-Xp22.3. AB - Key features of the oral-facial-digital syndrome type 1 (OFD1) include malformations of the face, oral cavity and digits. In addition, the clinical phenotype often includes mental retardation and renal functional impairment. Approximately 75% of cases of OFD1 are sporadic, and the condition occurs almost exclusively in females. In familial cases, the most likely mode of inheritance is considered to be X-linked dominant with prenatal lethality in affected males. Therefore, the OFD1 gene product appears to have widespread importance in organogenesis and is essential for fetal survival. We have studied two kindreds in which the clinical course was dominated by polycystic kidney disease requiring dialysis and transplantation. Using polymorphic chromosome markers spaced at approximately 10 cM intervals along the X chromosome, we mapped the disease to a region on the short arm of the X chromosome (Xp22.2-Xp22.3) spanning 19.8 cM and flanked by crossovers with the markers DXS996 and DX7S105. There was a maximum lod score of 3.32 in an 'affecteds only' analysis using a marker within the KAL gene (theta = 0.0 ), thereby confirming the location of the gene for OFD1 on the X chromosome. The remainder of the X chromosome was excluded by recombinants in affected individuals. The importance of our findings includes the definitive assignment of this male-lethal disease to the X chromosome and the mapping of a further locus for a human polycystic kidney disease. Furthermore, this mapping study suggests a possible mouse model for OFD1 as the X-linked dominant Xpl mutant, in which polydactyly and renal cystic disease occurs, maps to the homologous region of the mouse X chromosome. PMID- 9215689 TI - Germline mutations of the MEN1 gene in familial multiple endocrine neoplasia type 1 and related states. AB - Familial multiple endocrine neoplasia type 1 (FMEN1) is an autosomal dominant trait characterized by tumors of the parathyroids, gastro-intestinal endocrine tissue, anterior pituitary and other tissues. We recently cloned the MEN1 gene and confirmed its identity by finding mutations in FMEN1. We have now extended our mutation analysis to 34 more unrelated FMEN1 probands and to two related states, sporadic MEN1 and familial hyperparathyroidism. There was a high prevalence of heterozygous germline MEN1 mutations in sporadic MEN1 (8/11 cases) and in FMEN1 (47/50 probands). One case of sporadic MEN1 was proven to be a new MEN1 mutation. Eight different mutations were observed more than once in FMEN1. Forty different mutations (32 FMEN1 and eight sporadic MEN1) were distributed across the MEN1 gene. Most predicted loss of function of the encoded menin protein, supporting the prediction that MEN1 is a tumor suppressor gene. No MEN1 germline mutation was found in five probands with familial hyperparathyroidism, suggesting that familial hyperparathyroidism often is caused by mutation in another gene or gene(s). PMID- 9215690 TI - Identification of the multiple endocrine neoplasia type 1 (MEN1) gene. The European Consortium on MEN1. AB - Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant disorder characterised by tumours of the parathyroids, pancreas and anterior pituitary that represents one of the familial cancer syndromes. The MEN1 locus has been previously localised to chromosome 11q13, and a <300 kb gene-rich region flanked centromerically by PYGM and telomerically by D11S1783 defined by combined meiotic and tumour deletion mapping studies. Two candidate genes, ZFM1 and PPP2R5B, from this region have been previously excluded, and in order to identify additional candidate genes we used a BAC to isolate cDNAs from a bovine parathyroid cDNA library by direct selection. One of the novel genes that we identified, SCG2, proved to be identical to the recently published MEN1 gene, which is likely to be a tumour suppressor gene. The SCG2 transcript was 2.9 kb in all tissues with an additional 4.2 kb transcript also being present in the pancreas and thymus. Mutational analysis of SCG2 in 10 unrelated MEN1 families identified one polymorphism and nine different heterozygous mutations (one missense, four non sense, one insertional and three deletional frameshifts) that segregated with the disease, hence providing an independent confirmation for the identification of the MEN1 gene. PMID- 9215691 TI - Dystrobrevin deficiency at the sarcolemma of patients with muscular dystrophy. AB - Mutations in the genes encoding dystrophin or dystrophin-associated proteins are responsible for Duchenne muscular dystrophy or various forms of limb-girdle muscular dystrophies respectively. We have recently cloned the gene for the murine 87 kDa postsynaptic protein dystrobrevin, a dystrophin-associated protein. Anti-dystrobrevin antibodies stain the sarcolemma in normal skeletal muscle indicating that dystrobrevin co-localises with dystrophin and the dystrophin associated protein complex. By contrast, dystrobrevin membrane staining is severely reduced in muscles of Duchenne muscular dystrophy patients, consistent with dystrobrevin being a dystrophin-associated protein. Interestingly, dystrobrevin staining at the sarcolemma is dramatically reduced in patients with limb-girdle muscular dystrophy arising from the loss of one or all of the sarcoglycan components. Normal dystrobrevin staining is observed in patients with other forms of limb-girdle muscular dystrophy where dystrophin and the rest of the dystrophin-associated protein complex are normally expressed and in other neuromuscular disorders. Our results show that dystrobrevin-deficiency is a generic feature of dystrophies linked to dystrophin and the dystrophin-associated proteins. This is the first indication that a cytoplasmic component of the dystrophin-associated protein complex may be involved in the pathogenesis of limb girdle muscular dystrophy. PMID- 9215692 TI - Cloning and characterization of a neural cell recognition molecule on axons of the retinotectal system and spinal cord. AB - Immunoglobulin superfamily molecules in the brain are involved in distinct aspects of nervous system histogenesis, for example neuronal migration and axonal growth. To identify novel members of this superfamily in the chick nervous system, we developed a polymerase chain reaction-based approach making use of sequence motifs of immunoglobulin-like domains. In the present study, we report the molecular cloning of three isoforms, the biochemical analysis and the immunohistochemical characterization of one of the proteins identified in this screen. This molecule has 91% sequence identity with the limbic system-associated membrane protein (LAMP) characterized in the rat and is therefore referred to as the chicken homologue of the latter (chLAMP). The molecule is a glycosylphosphatidyl-inositol-anchored 60 kDa protein with three immunoglobulin like domains and contains 40% N-linked carbohydrate. We identify three different mRNA forms of chLAMP and show that two forms with distinct 5'-termini are differentially transcribed in neural development. In addition, we demonstrate using a fusion protein expressed in eukaryotic cells that chLAMP has homophilic binding activity. The protein was found on a subset of axons in the central and peripheral nervous system and is likely to be involved in specific cell-cell interactions in neurohistogenesis. PMID- 9215693 TI - Increased intermale aggression and neuroendocrine response in mice deficient for the neural cell adhesion molecule (NCAM). AB - Mice deficient for the neural cell adhesion molecule (NCAM) show morphological and behavioural abnormalities in the adult form, including a reduced size of the olfactory bulb, reduced exploratory behaviour, and deficits in spatial learning. Here we report increased aggressive behaviour of both homozygous (NCAM -/-) and heterozygous (NCAM +/-) male mutant mice towards an unfamiliar male intruding into their home cage. While plasma testosterone concentrations did not differ between genotypes before or after behavioural testing, corticosterone levels were higher in mutant residents than in wild-type (NCAM +/+) residents 30 min after encountering the intruder. Levels of c-fos mRNA, analysed to monitor neuronal activation, were similar in primary output structures of the olfactory bulb in NCAM-deficient and NCAM +/+ mice, but were increased in brain areas of the limbic system in both NCAM -/- and NCAM +/- mutant mice after the behavioural test. These results indicate that abnormalities in social behaviour correlate with enhanced neuronal activity in limbic brain areas and result in increased social stress in NCAM-deficient mice. PMID- 9215694 TI - Effects of GDNF on axotomized sensory and motor neurons in adult rats. AB - Glial cell-line-derived neurotrophic factor (GDNF) is a potent neurotrophic factor shown to rescue developing and adult motoneurons in vitro and in vivo from programmed and injury-induced cell death. To test whether GDNF would rescue adult mammalian sensory and motor neurons from physiological consequences of injury, the tibial nerve of rats was axotomized and, after a 10 day delay to permit injury processes to proceed, vehicle or GDNF was supplied directly to the nerve for 2 or 4 weeks or GDNF intrathecally for 2 weeks. Conduction velocity (CV) of both sensory and motor axons declined during the initial 10 days, and even more so if then treated with vehicle. Treatment with GDNF resulted in marginal improvement of sensory axon CV. CV of motor axons recovered significantly in a dose- and time-dependent manner. The results suggest that GDNF may have therapeutic potential in the treatment of peripheral neuropathies. PMID- 9215695 TI - Cocaine-induced increase in cortical acetylcholine release: interaction with the hypothalamo-pituitary-adrenal axis. AB - An influence on drug-taking behaviours of the stress-related hypothalamo pituitary-adrenal (HPA) axis and its final hormonal mediator, corticosterone, has previously been demonstrated. A role for cortically projecting cholinergic neurons in these behaviours can also be proposed. The experiments presented here examine the effect of the drug of abuse cocaine (15 mg/kg) on the release of acetylcholine (ACh) in the cortex of freely moving rats, using the technique of in vivo microdialysis. To assess a possible modulatory influence of the HPA axis via its final hormonal mediator corticosterone, the cocaine-induced effect on cortical ACh release in intact rats was compared to that in adrenalectomized (ADX) rats, which thus lacked their endogenous source of corticosterone, and in ADX rats in which the cocaine-induced corticosterone peak and/or the basal circadian concentrations of serum corticosterone were simulated by replacement treatments. The results reported here demonstrate that cortical ACh release is greatly increased by cocaine in intact rats; ADX prolongs the return to basal levels of cortical ACh, and the chronic replacement of circadian levels of corticosterone normalizes this effect. In contrast, during the plateau period of cocaine-induced increased cortical ACh release, where no effect of ADX is evident, rats with chronic replacement of corticosterone show an attenuated cocaine-induced cortical ACh release, and the acute replacement of the cocaine induced corticosterone secretion further attenuates this response. These results demonstrate that cocaine stimulates cortically projecting cholinergic neurons, and that the HPA hormone corticosterone modulates this interaction in a complex manner which merits further investigation. PMID- 9215696 TI - GABAergic transcallosal neurons in developing rat neocortex. AB - In the mature cerebral cortex the interhemispheric connections across the corpus callosum appear to be essentially completely excitatory on the basis of both immunocytochemical and electrophysiological studies. During late embryonic development, however, immunocytochemical staining reveals numerous GABA-positive fibres in the callosum, which later largely disappear. The origin of these fibres and whether they represent functional GABAergic neurons has not been established. In the present study we used a combination of retrograde labelling in vivo with electrophysiology and immunocytochemistry in cell culture to show that transiently at birth in rat pups a substantial number of transcallosal cortical cells are functional GABAergic neurons. Possible roles and fates for these neurons are discussed. PMID- 9215698 TI - Responses of cortical EEG-related basal forebrain neurons to brainstem and sensory stimulation in urethane-anaesthetized rats. AB - The basal forebrain can be considered to be a rostral extension of the ascending reticular activating system. A large number of neurons in the basal forebrain have been shown to display higher firing rates when low-voltage fast activity is present in the cortical EEG as opposed to states characterized by large slow waves in both unanaesthetized and anaesthetized animals. However, a smaller number of cells with increased discharge rate during slow waves was also observed in most of these studies. While it is likely that these two types of neurons have opposite roles in the regulation of cortical activation, it is not known how they respond to inputs from the brainstem or the periphery. In the present study, extracellular recordings were made in the basal forebrain of urethane anaesthetized rats. A total of 52 neurons were studied in which the firing rate was significantly higher during fast cortical EEG waves (F-cells), and 14 neurons in which activity was significantly greater during slow waves (S-cells). The two cell types responded differently to stimulation of the pedunculopontine tegmental nucleus (PPT) and dorsal raphe nucleus (DRN) with short (0.5-1 s) trains of pulses and to noxious sensory stimuli (tail pinch). These stimulations excited most F-cells (80-96%) and inhibited the majority of S-cells (55-67%). In the few F-cells that were inhibited by stimulation, the response varied with the background firing rate of the cell: the higher the firing rate at the time of stimulation, the higher the probability of observing an inhibitory response. In contrast, single electrical pulses delivered to the PPT and DRN excited the majority (72%) of both F- and S-cells. Previous in vitro studies have shown that the application of acetylcholine or serotonin has predominantly inhibitory effects on basal forebrain cholinergic neurons. The predominantly excitatory effect of noxious, PPT and DRN stimulation on F-cells therefore suggests that glutamatergic or other excitatory afferents play a more dominant role in regulating basal forebrain neurons. We have previously shown that F-cells are more prevalent than S-cells. In combination, these findings suggest that basal forebrain neurons, and F-cells in particular, are important in mediating the ascending excitatory drive from the brainstem to the cerebral cortex. PMID- 9215697 TI - Inhibition of [Ca2+]i transients in rat adrenal chromaffin cells by neuropeptide Y: role for a cGMP-dependent protein kinase-activated K+ conductance. AB - The effects of neuropeptide Y on the intracellular level of Ca2+ ([Ca2+]i) were studied in cultured rat adrenal chromaffin cells loaded with fura-2. A proportion (16%) of cells exhibited spontaneous rhythmic [Ca2+]i oscillations. In silent cells, oscillations could be induced by forskolin and 1,9-dideoxyforskolin. This action of forskolin was not modified by H-89, an inhibitor of protein kinase A. Spontaneous [Ca2+]i fluctuations and [Ca2+]i fluctuations induced by forskolin- and 1,9-dideoxyforskolin were inhibited by neuropeptide Y. Increases in [Ca2+]i induced by 10 and 20 mM KCI but not by 50 mM KCI were diminished by neuropeptide Y. However, neuropeptide Y had no effect on [Ca2+]i increases evoked by (-)BAY K8644 and the inhibitory effect of neuropeptide Y on responses induced by 20 mM KCI was not modified by omega-conotoxin GVIA, consistent with neither L- nor N type voltage-sensitive Ca2+ channels being affected by neuropeptide Y. Rises in [Ca2+]i provoked by 10 mM tetraethylammonium were not decreased by neuropeptide Y, suggesting that K+ channel blockade reduces the effect of neuropeptide Y. However, [Ca2+]i transients induced by 1 mM tetraethylammonium and charybdotoxin were still inhibited by neuropeptide Y, as were those to 20 mM KCI in the presence of apamin. The actions of neuropeptide Y on [Ca2+]i transients provoked by 20 and 50 mM KCI, 1 mM tetraethylammonium, (-)BAY K8644 and charybdotoxin were mimicked by 8-bromo-cGMP. In contrast, 8-bromo-cAMP did not modify responses to 20 mM KCI or 1 mM tetraethylammonium. The inhibitory effects of neuropeptide Y and 8-bromo-cGMP on increases in [Ca2+]i induced by 1 mM tetraethylammonium were abolished by the Rp-8-pCPT-cGMPS, an inhibitor of protein kinase G, but not by H 89. A rapid, transient increase in cGMP level was found in rat adrenal medullary tissues stimulated with 1 microM neuropeptide Y. Rises in [Ca2+]i produced by DMPP, a nicotinic agonist, but not by muscarine, were decreased by neuropeptide Y. Our data suggest that neuropeptide Y activates a K+ conductance via a protein kinase G-dependent pathway, thereby opposing the depolarizing action of K+ channel blocking agents and the associated rise in [Ca2+]i. PMID- 9215699 TI - PrP and beta-amyloid fragments activate different neurotoxic mechanisms in cultured mouse cells. AB - Alzheimer's disease and prion diseases such as Creutzfeldt-Jakob disease are caused by as yet undefined metabolic disturbances of normal cellular proteins, the amyloid precursor protein and the prion protein (PrP). Synthetic fragments of both proteins, beta-amyloid 25-35 (betaA25-35) and PrP106-126, have been shown to be toxic to neurons in culture. Cell death in both cases occurs by apoptosis. Here we show that there are considerable differences in the mechanisms involved. Thus, PrP106-126 is not toxic to cortical cell cultures of PrP knockout mouse neurons whereas betaA25-35 is. The toxicity of both peptides involves Ca2+ uptake through voltage-sensitive Ca2+ channels but only PrP106-126 toxicity involves the activity of NMDA receptors. The toxicity of betaA25-35, but not PrP106-126, is attenuated by the action of forskolin. These results indicate that PrP106-126 and PA25-35 induce neuronal apoptosis through different mechanisms. PMID- 9215700 TI - Heparin inhibits acetylcholine receptor aggregation at two distinct steps in the agrin-induced pathway. AB - Muscle cells depend on motoneurons for the initiation of postsynaptic differentiation during early development of the neuromuscular junction. Motoneurons secrete specific isoforms of the extracellular matrix protein agrin which trigger the aggregation of acetylcholine receptors (AChRs) on the muscle surface. Both motoneuron- and agrin-induced AChR aggregation are inhibited by heparin. Here we show that this inhibition is due to two separate and distinguishable mechanisms. At high concentrations, heparin directly binds to agrin isoforms which contain the peptide KSRK, resulting in a virtually complete inhibition of AChR clustering. Heparin and other polyanions do not bind to agrin splicing variants without KSRK insert. Isoforms containing or lacking the KSRK insert have a high potency to induce AChR aggregation in the presence of an activating eight-amino-acid insert. This activity is inhibited by low concentrations of heparin even in the absence of any binding of heparin to agrin. Therefore, this second type of inhibition is due to the interaction of heparin with a downstream component of the agrin-induced clustering pathway. Binding of heparin to this yet unidentified component substantially decreases, but does not completely abolish AChR aggregation. The inhibition is particularly strong on myotubes which have not completely matured in culture. PMID- 9215701 TI - Relative changes in regional cerebral blood flow during spinal cord stimulation in patients with refractory angina pectoris. AB - Spinal cord stimulation applied at thoracic level 1 (T1) has a neurally mediated anti-anginal effect based on anti-ischaemic action in the myocardium. Positron emission tomography was used to study which higher brain centres are influenced by spinal cord stimulation. Nine patients with a spinal cord stimulator for angina pectoris were studied using H(2)(15)O as a flow tracer. Relative changes in regional cerebral blood flow related to stimulation compared with non stimulation were assessed and analysed using the method of statistical parametric mapping. Increased regional cerebral blood flow was observed in the left ventrolateral periaqueductal grey, the medial prefrontal cortex [Brodmann area (BA) 9/10], the dorsomedial thalamus bilaterally, the left medial temporal gyrus (BA 21), the left pulvinar of the thalamus, bilaterally in the posterior caudate nucleus, and the posterior cingulate cortex (BA 30). Relative decreases in rCBF were noticed bilaterally in the insular cortex (BA 20/21 and BA 38), the right inferior temporal gyrus (BA 19/37), the right inferior frontal gyrus (BA 45), the left inferior parietal lobulus (BA 40), the medial temporal gyrus (BA 39) and the right anterior cingulate cortex (BA 24). It is concluded that spinal cord stimulation used as an additional treatment for angina applied at T1 modulates regional cerebral blood flow in brain areas known to be associated with nociception and in areas associated with cardiovascular control. PMID- 9215702 TI - Induction of adhesion/differentiation of human neuroblastoma cells by tumour necrosis factor-alpha requires the expression of an inducible nitric oxide synthase. AB - Neuroblastoma cell lines (SK-N-SH and SK-N-MC) were induced to differentiate, as detected by the expression of neurofilament proteins of 68 and 200 kDa, and to express adhesion molecules (intercellular adhesion molecule-1 and vascular cell adhesion molecule) after stimulation with tumour necrosis factor-alpha (TNF alpha). This induction was accompanied by the arrest of cell growth. The induction of neuroblastoma adhesion by TNF-alpha could be inhibited by the nitric oxide synthase inhibitors, L-N-monomethyl arginine (L-NMMA) and L-N6-(1 imidoethyl)-lysine (highly specific for the inducible enzyme), but not by the inactive enantiomer D-NMMA. These results indicate that TNF-alpha induces the adhesion of neuroblastoma cells via nitric oxide. This was confirmed by the finding that the adhesion/differentiation of SK-N-SH and SK-N-MC cells can be directly induced by the addition of nitric oxide donors, sodium nitroprusside and S-nitroso-N-acetyl-penicillamine, into the culture medium. The isoform of the nitric oxide synthase induced in human neuroblastoma cells by TNF-alpha treatment was identified enzymatically as isoform II by Western blotting and by the polymerase chain reaction. Thus TNF-alpha induces the in vitro adhesion/differentiation of human neuroblastoma cells through nitric oxide synthesized by a calcium-independent inducible form of nitric oxide synthase, clearly indicating that isoform II of nitric oxide synthase can be expressed in human neuronal cell types. PMID- 9215703 TI - Gx/z is regulated by mu but not delta opioid receptors in the stimulation of the low Km GTPase activity in mouse periaqueductal grey matter. AB - High affinity low K(m) GTPase activity was measured in membrane preparations of adult mouse mesencephalic periaqueductal grey matter (PAG). Opioids displaying selectivity towards mu- or delta-opioid receptors (OR) activated the enzyme in a concentration-dependent manner. Antibodies to mu-OR greatly impaired the potential of mu-agonists, [D-Ala2,N-MePhe4,Gly-ol5]-enkephalin (DAMGO) and morphine, to increase hydrolysis of GTP. The same antibodies had little effect on [D-Pen2,5]enkephalin (DPDPE) and [D-Ala2]deltorphin II, both agonists at delta OR. Stimulation of GTPase by DPDPE and [D-Ala2]deltorphin II - but not by morphine or DAMGO - was diminished by antibodies to delta-OR. The blockade of G(i2)alpha subunits by specific antibodies impaired the activation of G alpha related GTPase by all opioids. Antibodies in vitro, and oligodeoxynucleotides in vivo, prepared against Gx/z alpha subunits, reduced the release of Pi promoted by DAMGO and morphine. The impairment of Gx/z proteins also slightly reduced the effect of the delta2 agonist [D-Ala2]deltorphin II. At delta1 receptors, DPDPE fully expressed its activation of GTPase. These results indicate that in the PAG, mu-OR and delta-OR couple with Gi2 transducer proteins. Notably, mu-OR also regulates the pertussis toxin-insensitive G-protein Gx/z, an effect poorly exhibited by delta-OR in this tissue. PMID- 9215704 TI - Alpha7 and alpha8 nicotinic receptor subtypes immunopurified from chick retina have different immunological, pharmacological and functional properties. AB - Nicotinic receptors are present in the chick retina, but their structure and functional characteristics are still unclear. Using anti-alpha7 and anti-alpha8 subunit-specific antibodies, we immunopurified the alpha7 and alpha8 subtypes of chick retina neuronal nicotinic receptors. When analysed by sodium dodecyl sulphate-polyacrylamide gel electrophoresis, the two purified subtypes consistently showed a similar peptide composition characterized by the presence of two major peptides of M(r) 58 +/- 1 and 54 +/- 1 kDa, and two minor peptides of 67 and 61 +/- 1 kDa. In the alpha7 subtype, the 58 kDa peptide was recognized by anti-alpha7 but not by anti-alpha8 antibodies; in the alpha8 subtype, the 58 kDa peptide was recognized only by anti-alpha8 antibodies. The alpha7 subtype had a single class of [125I]alpha-bungarotoxin binding sites with a K(D) value of 1.2 nM, whereas the purified alpha8 subtype had two classes of binding sites, one with a K(D) of 5.5 nM and the other with very high affinity (KD 52 pM), but present in only 8% of the receptors. Competition binding experiments also showed the presence on the alpha8 subtype of high- and low-affinity classes of binding sites; the affinity for cholinergic drugs of the former was greater than that of the single class present on the alpha7 subtype. When reconstituted in planar lipid bilayers, both subtypes formed ligand-gated cation channels with major conductance levels of 42 and 52 pS but with different lifetimes; the two channels were activated by agonists and blocked by d-tubocurarine and the glycinergic antagonist strychnine. In line with the binding data, the reconstituted alpha8 subtype had greater agonist sensitivity than the alpha7 subtype. PMID- 9215705 TI - Localization of neuropeptide Y Y1 mRNA in the human brain: abundant expression in cerebral cortex and striatum. AB - Many neurobiological functions have been ascribed to the NPY Y1 receptor subtype, but autoradiographic analysis has failed to detect Y1 binding sites in most human brain areas, in contrast to the rat. We examined the regional distribution of Y1 mRNA-containing cells in the post-mortem human brain to clarify if there is a major species difference in terms of the existence of Y1 receptors in the human telencephalon, in particular the striatum and cortex. In situ hybridization experiments revealed widespread distribution of Y1 mRNA signals in all layers of most limbic and neocortical regions, predominantly in layer IV (most cortical regions) and layer VI. The striatum showed moderate Y1 receptor mRNA expression levels with intensely expressing cells localized to the nucleus accumbens. The highest Y1 receptor mRNA expression was apparent within the dentate gyrus, and the lowest in the subiculum, parahippocampal gyrus, cerebellum, and thalamus. In vitro autoradiography using [125I]Leu31Pro34-PYY and [125I]PYY with NPY (13-36) or Leu31Pro34-NPY, confirmed the presence of low Y1-like binding in the human brain despite abundant Y1 mRNA expression. However, using a rat model of the human autopsy process, it was apparent that the inability to reveal high Y1- versus Y2-like receptors in the human brain was related in part to marked reductions of Y1-like, but not Y2-like, receptors within a 4 h post-mortem delay. Altogether, the results indicate that the Y1 receptor gene is abundant in the human brain and this receptor may have important roles in cognitive, limbic and motor function. PMID- 9215706 TI - The whole nucleotide sequence and chromosomal localization of the gene for human metabotropic glutamate receptor subtype 6. AB - Metabotropic glutamate receptor subtype 6 (mGluR6) is restrictedly expressed in the retinal ON bipolar cells and ablation of mouse mGluR6 by gene targeting results in a loss of ON responses to light stimulus and impairs the detection of visual contrasts. We have isolated genomic clones containing the human mGluR6 gene and determined the whole nucleotide sequence of the mGluR6 gene. The transcription initiation site of the human mGluR6 gene has been identified using primer extension analysis in combination with reverse transcriptase-mediated polymerase chain reaction analysis of human retinal RNA, while the termination of the mGluR6 mRNA has been assigned by the analysis of rapid amplification of 3' cDNA ends. The human mGluR6 gene consists of 16,742 base pairs with 10 exons separated by nine introns. The human mGluR6 is composed of 877 amino acid residues with a signal peptide of 24 amino acid residues and the mature protein shows a 94.6% homology with the rat counterpart. A CpG-rich island is present at exon 1 and its preceding putative promoter region and this unusual sequence, like several tissue-specific genes, may be important for a specific expression of the mGluR6 gene in the retinal bipolar cells. The human mGluR6 gene has been mapped to chromosome 5q35 by the analyses of blot hybridization of a DNA panel of human/mouse/hamster somatic cell hybrids and fluorescence in situ hybridization of human chromosomes. This study should provide the genetic basis for not only better understanding the molecular mechanism underlying a tissue-specific expression of the mGluR6 gene but also exploring a potential defect in human mGluR6 in a certain inherited eye disease. PMID- 9215707 TI - Neuronal and glial glutamate transporters possess an SH-based redox regulatory mechanism. AB - Glutamate uptake into nerve cells and astrocytes via high-affinity transporters controls the extracellular glutamate concentration in the brain, with major implications for physiological excitatory neurotransmission and the prevention of excitotoxicity. We report here that three recently cloned rat glutamate transporter subtypes, viz. EAAC1 (neuronal), GLT1 and GLAST (glial), possess a redox-sensing property, undergoing opposite functional changes in response to oxidation or reduction of reactive sulphydryls present in their structure. In particular, thiol oxidation with 5,5'-dithio-bis(2-nitrobenzoic) acid (DTNB) and disulphide reduction with dithiothreitol (DTT) result, respectively, in reduced and increased uptake capacity by a preparation of partially purified brain transporters as well as by the three recombinant proteins reconstituted into liposomes. In this model system, EAAC1, GLT1 and GLAST react similarly to DTT/DTNB exposures despite their different contents of cysteines, suggesting that only the conserved residues might be involved in redox modulation. Redox sensitivity is a property of the glutamate transporters also when present in their native cell environment. Thus, by using cultured cortical astrocytes and the whole-cell patch-clamp technique we were able to observe dynamic increase and decrease of the glutamate uptake current in response to application of DTT and DTNB in sequence. Moreover, in the same paradigm, DDT-reversible current inhibition was observed with hydrogen peroxide instead of DTNB, indicating that the SH-based redox modulatory site is targeted by endogenous oxidants and might constitute an important physiological or pathophysiological regulatory mechanism of glutamate uptake in vivo. PMID- 9215708 TI - Leukaemia inhibitory factor is retrogradely transported by a distinct population of adult rat sensory neurons: co-localization with trkA and other neurochemical markers. AB - Sciatic sensory afferents that retrogradely transport and accumulate leukaemia inhibitory factor (LIF) within their soma were characterized in the adult rat in vivo. Twenty-four percent of neurons within the L4 and L5 dorsal root ganglia accumulated biotinylated LIF following intraneural injection of the cytokine into the sciatic nerve. Labelled cell bodies were predominantly of small diameter (20.1 +/- 0.5 microm). Retrograde transport was eliminated by excess unlabelled LIF but not by the related cytokines, ciliary-derived neurotrophic factor (CNTF) and interleukin-6 (IL-6). Double labelling revealed that the majority (81%) of LIF-accumulating neurons were immunopositive for CGRP and 34% were immunopositive for the cell surface glycoconjugate IB4. Sixty-two percent of LIF-accumulating neurons were immunopositive for trkA. Our results demonstrate a group of small diameter sensory neurons that retrogradely transport LIF, comprising cells that constitutively express neuropeptides and those likely to be peptide-deficient. LIF-accumulating neurons expressing trkA are also potentially responsive to nerve growth factor. It is likely that the LIF-accumulating neurons described in this study are nociceptive in function. PMID- 9215709 TI - Regulation of rat neuronal voltage-dependent calcium channels by endogenous p21 ras. AB - Influx of calcium through voltage-dependent calcium channels (VDCCs) has been implicated in the processes of cell growth and differentiation. Various signalling proteins, including nerve growth factor (NGF), p21-ras and src tyrosine kinases, have been suggested to have a role in the regulation of neuronal VDCCs. Using the whole-cell patch-clamp technique we have investigated the role of endogenous p21-ras in the regulation of VDCCs in primary cultured dorsal root ganglion (DRG) neurons obtained from neonatal rats. Neutralization of endogenous p21-ras by microinjection of p21-ras antibody (Y13-259) reduced the maximum peak barium current, I(max), whereas microinjection of oncogenic p21-K ras increased the current. Thus, endogenous p21-ras is involved in the tonic regulation of calcium currents in these cells. Intracellular application of a phosphopeptide, Trk 490, which prevents the binding of the adaptor protein shc to the activated NGF receptor, so blocking p21-ras activation, reduced I(max). Similarly, deprivation of NGF by overnight incubation in NGF-free medium also reduced I(max). Together, these results suggest that NGF receptor tyrosine kinase activation of p21-ras is likely to be involved in the tonic regulation of VDCCs in DRG neurons. Deprivation of NGF combined with microinjection of p21-ras antibody (Y13-259), however, caused an even greater reduction of I(max). Thus, NGF activation can only partially explain the regulation of these currents by endogenous p21-ras. Src tyrosine kinases have been suggested to activate p21-ras. In DRG neurons, microinjection of purified src tyrosine kinase, pp60c-src, increased I(max) in these cells. However, co-microinjection of pp60c-src with Y13 259 antibody prevented the increase in I(max), implying that pp60c-src can also regulate calcium currents via the activation of endogenous p21-ras. Further support for the involvement of tyrosine kinases in VDCC regulation was provided by the application of the general tyrosine kinase inhibitor, genistein, which also reduced I(max). Thus, VDCCs in rat DRG neurons appear to be tonically up regulated by endogenous p21-ras. This effect appears largely to involve NGF receptor tyrosine kinase activation of p21-ras. In addition, src tyrosine kinase may also regulate VDCCs, possibly via p21-ras. PMID- 9215710 TI - Role of the dorso-caudal neostriatum in filial imprinting of the domestic chick: a pharmacological and autoradiographical approach focused on the involvement of NMDA-receptors. AB - Newly hatched domestic chicks were either acoustically imprinted on 400 Hz tone pulses or visually imprinted on a rotating red light. Compared to naive control animals, both groups of imprinted chicks expressed significantly enhanced stimulus evoked 2-fluoro-2-deoxyglucose (2-FDG) uptake in circumscribed areas of the dorso-caudal neostriatum (Ndc). This enhanced excitability after imprinting seems not to be related to changes of NMDA-receptor densities as measured by quantitative receptor autoradiography. However, pharmacological blockade of NMDA receptors in the dorso-caudal neostriatum leads to a marked suppression of stimulus-evoked 2-FDG uptake in the dorso-caudal neostriatum and also in the interconnected imprinting relevant forebrain area, medio-rostral neostriatum/hyperstriatum ventrale (MNH). Furthermore, chicks which received bilateral Ndc injections of the competitive NMDA antagonist DL-2-amino-5 phosphono valeric acid (APV) during the imprinting experiments showed a dose dependent decrease of imprinting success compared to vehicle-injected controls. These results indicate that the dorso-caudal neostriatum may represent a polysensory associative brain region in which visual and acoustic features of imprinting objects may be integrated. The activation in this area evoked by the imprinting stimulus during and after imprinting is critically dependent on NMDA receptor activation, which appears to be required for this learning process. PMID- 9215711 TI - Na(+)-Ca2+ exchange currents in cortical neurons: concomitant forward and reverse operation and effect of glutamate. AB - Na(+)-Ca2+ exchanger-associated membrane currents were studied in cultured murine neocortical neurons, using whole-cell recording combined with intracellular perfusion. A net inward current specifically associated with forward (Na+(o) Ca2+(i)) exchange was evoked at -40 mV by switching external 140 mM Li+ to 140 mM Na+. The voltage dependence of this current was consistent with that predicted for 3Na+:1Ca2+ exchange. As expected, the current depended on internal Ca2+, and could be blocked by intracellular application of the exchanger inhibitory peptide, XIP. Raising internal Na+ from 3 to 20 mM or switching the external solution from 140 mM Li+ to 30 mM Na+ activated outward currents, consistent with reverse (Na+(i)-Ca2+(o)) exchange. An external Ca2(+)-sensitive current was also identified as associated with reverse Na(+)-Ca2+ exchange based on its internal Na+ dependence and sensitivity to XIP. Combined application of external Na+ and Ca2+ in the absence of internal Na+ triggered a 3.3-fold larger inward current than the current activated in the presence of 3 mM internal Na+, raising the intriguing possibility that Na(+)-Ca2+ exchangers might concurrently operate in both the forward and the reverse direction, perhaps in different subcellular locations. With this idea in mind, we examined the effect of excitotoxic glutamate receptor activation on exchanger operation. After 3-5 min of exposure to 100-200 microM glutamate, the forward exchanger current was significantly increased even when external Na+ was reduced to 100 mM, and the external Ca2(+) activated reverse exchanger current was eliminated. PMID- 9215712 TI - FGF-2 induces nerve growth factor expression in cultured rat hippocampal neurons. AB - Basic fibroblast growth factor (FGF-2) is expressed in the hippocampus and has been demonstrated to promote neurotrophic effects on hippocampal neurons in vitro. We show that these neurons, even at the embryonic stage, express the mRNAs encoding the FGF receptors, bek and flg. We have characterized the effects of FGF 2 on the expression of nerve growth factor (NGF) using the reverse transcription coupled polymerase chain reaction, in situ hybridization and immunocytochemistry. In hippocampal neurons grown in the absence of serum, FGF-2 exposure induces an important elevation of NGF mRNA expression followed by a marked increase in NGF immunoreactivity. Combining in situ hybridization with an NGF probe and microtubule-associated protein-2 (MAP2) immunocytochemistry we show that the induction of NGF mRNA by FGF-2 is localized in MAP2-immunoreactive neurons. These results suggest roles for FGF-2 in the development of hippocampal neurons and in the maintenance of connections in the central nervous system, particularly the septo-hippocampal pathway, via the regulation of an important neurotrophin. PMID- 9215713 TI - Figure-ground segregation at contours: a neural mechanism in the visual cortex of the alert monkey. AB - An important task of vision is the segregation of figure and ground in situations of spatial occlusion. Psychophysical evidence suggests that the depth order at contours is defined early in visual processing. We have analysed this process in the visual cortex of the alert monkey. The animals were trained on a visual fixation task which reinforced foveal viewing. During periods of active visual fixation, we recorded the responses of single neurons in striate and prestriate cortex (areas V1, V2, and V3/V3A). The stimuli mimicked situations of spatial occlusion, usually a uniform light (or dark) rectangle overlaying a grating texture of opposite contrast. The direction of figure and ground at the borders of these rectangles was defined by the direction of the terminating grating lines (occlusion cues). Neuronal responses were analysed with respect to figure-ground direction and contrast polarity at such contours. Striate neurons often failed to respond to such stimuli, or were selective for contrast polarity; others were non selective. Some neurons preferred a certain combination of figure-ground direction and contrast polarity. These neurons were rare both in striate and prestriate cortex. The majority of neurons signalled figure-ground direction independent of contrast polarity. These neurons were only found in prestriate cortex. We explain these responses in terms of a model which also explains neuronal signals of illusory contours. These results suggest that occlusion cues are used at an early level of processing to segregate figure and ground at contours. PMID- 9215714 TI - 192 IgG-saporin-induced loss of cholinergic neurons in the septum abolishes cholinergic sprouting after unilateral entorhinal lesion in the rat. AB - After unilateral lesion of the entorhinal cortex, cholinergic septohippocampal fibres are believed to sprout in the denervated outer molecular layer of the rat dentate gyrus. This cholinergic sprouting has been demonstrated by acetylcholinesterase (AChE) histochemistry, a method said selectively to label cholinergic septohippocampal fibres in the hippocampus. However, a recent report has questioned this concept, suggesting that AChE may not be an adequate marker to monitor cholinergic sprouting and that other, non-cholinergic axons sprouting after entorhinal cortex lesion cause the dense AChE-positive band in the denervated outer molecular layer. In order to determine the contribution of cholinergic septohippocampal fibres to the dense AChE band appearing after entorhinal cortex lesion, the neurotoxin 192 IgG-saporin, known to destroy cholinergic neurons in the basal forebrain selectively, was used. Rats received bilateral injections of 192 IgG-saporin into the lateral ventricles 3 weeks before entorhinal cortex lesion, simultaneously with entorhinal cortex lesion, or 8 weeks after entorhinal cortex lesion. Immunocytochemistry for choline acetyltransferase (ChAT) and in situ hybridization for ChAT mRNA demonstrated the loss of cholinergic neurons in the medial septum and diagonal band after 192 IgG saporin treatment. The cholinergic sprouting response in the molecular layer, as visualized with AChE histochemistry, was abolished in all animals treated with immunotoxin. These data indicate that the dense AChE band forming after entorhinal cortex lesion represents the sprouting of cholinergic septohippocampal fibres. PMID- 9215715 TI - A metabolic mapping study of orientation discrimination and detection tasks in the cat. AB - Increasing evidence suggests that a large number of distinct cortical areas and associated subcortical structures participate in the processing of visual information and that different aspects of visual scenes are evaluated in different areas. This necessitates identification of cortical and subcortical regions cooperating in particular visual tasks. Using the 2-deoxyglucose technique, we monitored the differential activation of areas in the cat visual cortex participating in an orientation discrimination and a detection task. Concordant with previous lesion studies, we found increased activity levels in area 17 in the discrimination condition relative to the detection condition. In addition, the 2-deoxyglucose technique revealed discrimination-related increased activations in the claustrum, the putamen and in parts of the anteromedial, anterolateral and posterolateral lateral suprasylvian visual areas. Regions activated differentially with the detection task comprised subdivisions of areas 17, 18, 19 and 21, posterior area 7 (7p), several areas of the posterior part of the middle and posterior suprasylvian sulcus, the pulvinar complex and the superior colliculus. These results show that the 2-deoxyglucose technique is useful to investigate cognitive brain functions, and that different sets of cortical and subcortical regions are activated during two visual tasks with similar visual stimulation. PMID- 9215716 TI - Axonal remodeling during postnatal maturation of CA3 hippocampal pyramidal neurons. AB - Anatomical substrates were investigated for local circuit hyperexcitability that occurs in the CA3 subfield of the rat hippocampus during postnatal week 2. A transient excess of excitatory local circuit connectivity was hypothesized to underlie this hyperexcitability. To test this hypothesis, recurrent excitatory axon arbors from single biocytin-filled CA3 pyramidal cells were reconstructed. Arbors were analyzed in segments of area CA3 comparable in size to in vitro minislice preparations, which were shown to reproduce the developmental hyperexcitability seen in intact slices during postnatal week 2. Segments were then adjusted for hippocampal growth, based on age-dependent changes in neuron density in stratum pyramidale. Axon arbors were found to be short and possessed very few branches during the first postnatal week. By the second postnatal week, arbors had undergone dramatic growth and were much longer and more complex in their branching patterns. By adulthood, a significant decrease in all measures of arbor length and complexity was observed. Following growth adjustment, measures of axon length and varicosity number during week 2 were not significantly different from that of adulthood. However, the number of axon branches decreased by 50%. These results suggest that, during early postnatal life, there is exuberant outgrowth of local CA3 recurrent axons, and with maturation these recurrent collaterals are remodeled. Short-ranging, profusely branched axons appear to be replaced by longer-ranging arbors that possess fewer branches. Maturational changes in the dendritic location rather than the number of early formed recurrent excitatory synapses may explain developmental hyperexcitability of the hippocampal CA3 subfield. PMID- 9215717 TI - PSA-NCAM and B-50/GAP-43 are coexpressed by specific neuronal systems of the adult rat mediobasal hypothalamus that exhibit remarkable capacities for morphological plasticity. AB - The present study was designed to determine whether the mediobasal hypothalamus of adult rats contains neurons that continue to coexpress the highly polysialylated neural cell adhesion molecule (PSA-NCAM) and B-50/GAP-43, two proteins coexpressed by virtually all of the neurons of the fetal and neonatal rat central nervous system. Confocal laser scanning microscopy combined with double- or triple-fluorescence immunostaining was used to identify the hypothalamic neurons that express high levels of both PSA-NCAM and B-50/GAP-43 and to study the possible modifications of their morphological organization following a surgical lesion through the mediobasal hypothalamus. In intact animals, PSA-NCAM and B-50/GAP-43 were found to be colocalized within numerous fibers projecting throughout the external layer of the median eminence that were immunoreactive for either gamma-aminobutyric acid (GABA) or tyrosine hydroxylase (TH). Three to 30 days after a lesion through this region, numerous regenerating axonal sprouts, triple-immunostained for PSA-NCAM, B-50/GAP-43, and either GABA or TH, were detected along the ventricular surface of, and throughout the perivascular layer of, the median eminence. Surprisingly, high levels of PSA-NCAM and B-50/GAP-43 were also associated with numerous supraependymal neurons that exhibited long ramified processes and were immunoreactive for GABA but TH negative. The use of the proliferation marker, 3H-thymidine, further indicated that the emergence of such supraependymal neurons after median eminence lesion was not related to the proliferation of preexisting quiescent cells. These data indicate that the mediobasal hypothalamus of the adult rat contains two neuronal systems, in which the continued coexpression of PSA-NCAM and B-50/GAP-43 is related to remarkable capacities for postlesional, morphological plasticity. PMID- 9215718 TI - Synchronized overproduction of AMPA, kainate, and NMDA glutamate receptors during human spinal cord development. AB - Quantitative receptor autoradiography was used to map the distribution in the developing human spinal cord of the three types of ionotropic glutamate receptors. N-methyl-D-Aspartate (NMDA) receptors were labeled with [3H]glutamate, kainic acid (KA) receptors were labeled with [3H]KA, and alpha-amino-3-hydroxy-5 methyl-4-isoxazole proprionate (AMPA) receptors were labeled with [3H]AMPA. In the adult, labeling of all three receptor subtypes is largely restricted to the substantia gelatinosa (SG) in the dorsal horn, with very low level labeling elsewhere in the spinal gray matter. In marked distinction, in late fetal life, high level ligand binding is seen throughout the spinal gray matter. In early postnatal life, binding sites diminish in all regions, but least so in the SG, until the adult pattern emerges. Thus a coordinated transient high level of ionotropic glutamate receptor expression occurs within the developing spinal cord. Saturation analysis of ligand binding shows that the affinity of [3H]KA and [3H]AMPA binding is not developmentally regulated. In contrast, the affinity of [3H]glutamate binding to the NMDA receptor in the fetal ventral horn is three fold greater than in the adult ventral horn. Thus, in addition to quantitative changes in glutamate receptor expression, qualitative changes occur in the expression of NMDA receptors during development. The distinct glutamate receptor phenotype of fetal and early postnatal spinal cord cells suggests that alterations in the excitable properties of these cells plays an important role in activity-dependent development and in susceptibility to excitotoxic injury. PMID- 9215719 TI - In situ hybridization histochemical localization of prodynorphin messenger RNA in the central nervous system of the rat. AB - The distribution of preprodynorphin messenger RNA-containing perikarya in the central nervous system of the rat was determined with in situ hybridization histochemistry using a 35S-labelled complementary RNA probe. All of the regions of the central nervous system reported by other investigators to contain perikarya that synthesize prodynorphin-derived peptides, except the pedunculopontine tegmental nucleus, the accessory trigeminal nucleus, and the ventral nucleus of the trapezoid body, also contained perikarya that synthesize preprodynorphin messenger RNA. However, the olfactory bulb, the anterior olfactory nucleus, the islands of Calleja, the CA1-CA3 fields of the hippocampus, the septohippocampal nucleus, the diagonal band of Broca, the basal and cortical amygdaloid nuclei, the entopeduncular nucleus, the subthalamic nucleus, the superior colliculus, the Edinger-Westphal nucleus, the dentate nucleus, the raphes linearis and pontis, the dorsal cochlear nucleus, the medial vestibular nucleus, the inferior olive, and the dorsal motor nucleus of the vagus nerve also contained preprodynorphin messenger RNA-synthesizing perikarya. These observations suggest that prodynorphin-derived peptides have a much more pervasive role in central nervous system function than previously suspected. However, before the physiological significance of these observations can be judged, it will be necessary to determine whether all of the novel sites of preprodynorphin messenger RNA synthesis are sites of prohormone synthesis and conventional processing. PMID- 9215720 TI - Pattern of expression of the serotonin2C receptor messenger RNA in the basal ganglia of adult rats. AB - The distribution of the serotonin (5-HT) receptor 5-HT2C mRNA was examined at the single-cell level with in situ hybridization histochemistry and emulsion autoradiography in the basal ganglia and mesolimbic system of adult rats, with focus on the pallidum and the substantia nigra, which receive striatal inputs and play a critical role in basal ganglia function. 5-HT2C receptor mRNA expression was always restricted to a subpopulation of neurons in the regions examined. In the neostriatum, labeled neurons were more numerous in the rostral nucleus accumbens than in the caudal nucleus accumbens and were more numerous in the ventral and ventrolateral caudate-putamen than in the dorsal caudate-putamen, where labeled neurons were restricted to isolated clusters. In striatal target areas, dense labeling in the entopeduncular nucleus (internal pallidum, direct striatal output pathway) contrasted with an absence of labeling in the globus pallidus (external pallidum, indirect striatal output pathway). Double-label in situ hybridization in the substantia nigra revealed coexpression of 5-HT2C receptor mRNA with glutamic acid decarboxylase but not with tyrosine hydroxylase mRNA, indicating that it was restricted to gamma-aminobutyric acid (GABA)ergic neurons. In this region, dense labeling for 5-HT2C mRNA was found in half of the neurons at middle and caudal levels of both the pars compacta and the pars reticulata, with little labeling rostrally. The data suggest that drugs acting on the 5-HT2C receptor could selectively affect discrete neuronal populations in the basal ganglia and mesolimbic systems and indicate a new level of neurochemical heterogeneity among GABAergic neurons of the substantia nigra. PMID- 9215721 TI - Afferent innervation of gastrointestinal tract smooth muscle by the hepatic branch of the vagus. AB - To survey the vagal hepatic branch afferent projections to and the terminal specializations in the gastrointestinal tract, male Sprague-Dawley rats were given subdiaphragmatic vagotomies, sparing only the common hepatic branch, and were injected with 3 microl of 8% wheat germ agglutinin-horseradish peroxidase in the left nodose ganglion. The nodose ganglia, the stomach, the first 8 cm of duodenum, and the cecum were prepared as wholemounts and were processed with tetramethyl benzidine. Hepatic afferent innervation of the ventral stomach consisted of one or more bundles entering at the lower esophageal sphincter and coursing to the forestomach, where they branched into distinct terminal fields. The only fibers on the dorsal forestomach were distal branches and terminals that wrapped around the greater curvature from the ventral side. Hepatic afferents supplied the forestomach with both intraganglionic laminar endings (IGLEs; putative mechanosensors that coordinate peristalsis) and intramuscular arrays (IMAs; considered tension receptors). IGLEs were located primarily on the ventral wall of the stomach, whereas IMAs were distributed symmetrically. Afferents were also supplied to the distal antrum and the pylorus, with pyloric innervation consisting almost exclusively of IMAs. Innervation of the proximal duodenum was denser in the first 3 cm and decreased progressively caudally, with only meager innervation after 6 cm. Cecal innervation consisted of a few fibers at the ileocecal junction. Duodenal and cecal endings were predominately IGLEs. These results indicate that the hepatic branch carries sensory information from the forestomach, antrum, pylorus, duodenum, and cecum. Furthermore, the different terminals it supplies suggest that the branch mediates a multiplicity of gastrointestinal functions. PMID- 9215722 TI - Immunoglobulin molecules are present in early-generated neuronal populations in the rat cerebral cortex and retina. AB - Immunoglobulin-like antigens have been identified in neuronal subsets restricted to deeper layers of the developing mammalian cerebral cortex. The pattern is suggestive of selective uptake of immunoglobulin (Ig) from serum or synthesis of Ig or a molecule with Ig-like motifs. To distinguish between these alternatives, biochemical, immunocytochemical, and birthdating analyses were conducted. In neonatal rat cerebral cortex, immunoglobulin-like immunoreactivity was chiefly in subplate neurons, marginal zone neuropil, and processes spanning the cortical plate. Isolated staining was also observed within the ventricular zone. Although most staining in the cortical plate was absent several days after birth, subplate neuron staining persisted until the end of the second postnatal week. Quantitative immunoassays showed that the antigen concentration dropped from 130 ng/mg in cortical cytosol at birth to approximately 80 ng/mg by postnatal day 7 (P7) and remained low thereafter. Two Ig-immunoreactive polypeptides with mobilities similar to heavy and light chains of serum IgG, were identified by Western blotting. The larger band was purified, partially sequenced by Edman degradation, and found to match rat IgG heavy chain. Bromodeoxyuridine birthdating and anti-IgG double-labeling studies showed that most of the Ig containing cells were early-generated neurons. Outside of the cortex, transient IgG staining was also detected in neurons of the retina and cerebellum. These studies suggest that subplate and other early-generated neurons selectively take up Ig from serum. The IgG may then either be degraded or lost from the central nervous system (CNS) during developmentally regulated cell death. PMID- 9215723 TI - Distribution of gonadotropin-releasing hormone immunoreactivity in the brain of Ichthyophis beddomei (Amphibia: Gymnophiona). AB - From a comparative viewpoint, we have investigated the presence and neuroanatomical distribution of gonadotropin-releasing hormone (GnRH) immunoreactive material in the brain of a gymnophione amphibian, Ichthyophis beddomei. Immunocytochemical analysis of the adult brain and terminal nerves in both sexes shows the presence of neurons and fibers containing mammalian GnRH (mGnRH)- and chicken GnRH-II (cGnRH-II)-like peptides. With respect to GnRH immunoreactive material, there are two distinct neuronal systems in the brain: one containing mGnRH, which is located in the forebrain and terminal nerve, and the other containing cGnRH-II, which is restricted to the midbrain tegmentum. Basically, this distribution pattern parallels that of many species of anurans and a urodele. Whereas the presence of cGnRH-II-immunoreactive fibers in the dorsal pallium of L. beddomei is a feature in common with a urodele amphibian, the total absence of cGnRH-II-like material in the median eminence is unique to this species. It is suggested here that the distribution profile of GnRH-like material within the brain and terminal nerve of I. beddomei represents a primitive pattern. PMID- 9215724 TI - Topography of spiral ganglion projections to cochlear nucleus during postnatal development in cats. AB - A fundamenntal organizational principle of the central auditory system is that virtually all areas are tonotopically organized. However, we know very little about the timing or mechanisms that are responsible for the development of this organization. When cats are born, their auditory nervous systems are extremely immature, and their hearing thresholds are very high. Until postnatal days 7-10 (P7-10), cats have behavioral and physiological thresholds which are near or above the pain threshold for adults and also have poor frequency selectivity. Physiological thresholds for auditory nerve fibers and cochlear nucleus neurons are typically above 100-120 dB SPL (sound pressure level re 20 microPa). Three weeks later (at approximately P31), the sensitivity and frequency discrimination (tuning) of these neurons approximate adult values. This study examines the development of the tonotopic projections from the spiral ganglion to the cochlear nucleus during the period in cat development in which the auditory system undergoes the transition from being essentially nonfunctional to having adult like function. With the animals heavily anesthetized, the cochleas were surgically exposed in kittens ranging in age from P6 to P45. Focal injections of Neurobiotin (NB) were made into Rosenthal's canal, labeling a small cluster of cells in the spiral ganglion of each cochlea. The projections of these labeled cells were visualized as frequency-specific bands of labeled axons and terminals in all major subdivisions of the cochlear nucleus. The thickness of these bands (i.e., the dimension of the bands orthogonal to the isofrequency representation and across the frequency gradient) were measured and compared to similar projections in adults. As in adult cats, the thickness of the bands varied only slightly with the location of the injection site (frequency representation) over a range of 1-7 mm from the cochlear base (45-13 kHz). Moreover, band thickness did not vary significantly with age. These data indicate that the tonotopic organization of spiral ganglion projections to the cochlear nucleus is as precise in kittens as young as P6 as it is in adults. PMID- 9215725 TI - Neocortical neuron number in humans: effect of sex and age. AB - Modern stereological methods provide precise and reliable estimates of the number of neurons in specific regions of the brain. We decided to estimate the total number of neocortical neurons in the normal human brain and to analyze it with respect to the major macro- and microscopical structural components, to study the internal relationships of these components, and to quantitate the influence of important physiological variables on brain structure. The 94 brains reported represent a consecutive collection of brains from the general Danish population. The average numbers of neocortical neurons were 19 billion in female brains and 23 billion in male brains, a 16% difference. In our study, which covered the age range from 20 years to 90 years, approximately 10% of all neocortical neurons are lost over the life span in both sexes. Sex and age were the main determinants of the total number of neurons in the human neocortex, whereas body size, per se, had no influence on neuron number. Some of the data presented have been analyzed by using new mathematical designs. An equation predicting the total neocortical neuron number in any individual in which sex and age are known is provided. PMID- 9215726 TI - Lectin-binding properties of oligodendrocyte lineage cells aid in defining functionally important surface proteins. AB - Oligodendrocytes are the myelin-forming cells of the central nervous system. They develop from migratory and proliferative precursor cells, which differentiate to mature myelinating cells. As a first step toward investigating the expression of cell surface glycoproteins by oligodendrocyte lineage cells, we tested 14 different lectins for their binding to oligodendrocyte lineage cells. Peanut agglutinin (PNA) was the only lectin used that showed a differentiation stage dependent binding to oligodendrocytes. PNA-binding molecules are specifically expressed by oligodendrocyte precursor cells, downregulated with differentiation, and reexpressed by mature oligodendrocytes. It was additionally observed that PNA stimulates the proliferation of oligodendrocyte precursor cells. PNA may therefore be a useful tool for isolating and characterizing important cell surface glycoproteins expressed by oligodendrocyte lineage cells. PMID- 9215727 TI - Expression of high-affinity neuronal and glial glutamate transporters in the rat optic nerve. AB - Recent studies have revealed that a dynamic axon-glial signaling occurs in the rat optic nerve, which is devoid of synapses. This interaction is postulated to be mediated by non-vesicular release of glutamate via a reversal of high-affinity glutamate transporters. Here we examined the expression of glial glutamate transporters (GLAST and GLT-1) and a neuronal transporter (EAAC1) in the rat optic nerve. RT-PCR analysis revealed the presence of mRNAs for GLT-1 and GLAST, but not EAAC1. RNase protection assays showed that of the two glial transporters, mRNA for GLAST was expressed at much higher level than was GLT-1. A similar expression pattern was found in primary astrocyte culture cells. GLAST mRNA level in the optic nerve was comparable to that in the cerebellum. Developmentally, GLAST mRNA level was highest at P2 and dropped slightly by adulthood. Nerve transection resulted in little or no change in mRNA levels for GLAST and GLT-1 assayed at 4 to 14 days post-transection, but GLAST mRNA level was decreased at 64 days. Western blot analysis revealed that the rat optic nerve showed immunoreactivity to antibodies against GLT-1, GLAST, and EAAC1. In conclusion, we suggest that glial and neuronal transporters are present in the rat optic nerve, where dynamic axon-glial interaction has been known to occur. In particular, the unusually high level of expression of GLAST in the optic nerve suggests a possible role for this glial transporter in protecting optic nerves from neurotoxicity during postnatal development. PMID- 9215728 TI - Glial cells assemble hyaluronan-based pericellular matrices in vitro. AB - The extracellular matrix (ECM) of the brain contains hyaluronan and proteoglycans, as does the ECM of cartilage. Aggrecan, the major proteoglycan of cartilage, forms large aggregates with hyaluronan, which then associate with the chondrocyte cell surface through an interaction with surface hyaluronan binding proteins. In culture, chondrocytes elaborate hyaluronan-proteoglycan aggregates, which form large hydrated pericellular matrices (PCMs) that can be visualized by a particle exclusion assay (Knudson and Toole: Dev Biol 112:308, 1985). It has recently been demonstrated that embryonic glial cells can also elaborate PCMs in culture (Deyst and Toole: Dev Brain Res 28:351, 1995). We demonstrate here that different classes of glial cells elaborate different types of endogenous PCMs in culture. Less differentiated glial cells, as evidenced by their immunoreactivity for nestin, elaborate larger endogenously produced PCMs than differentiated astrocytes, as defined by immunoreactivity for GFAP. This in vitro result may be a reflection of the larger volume of extracellular space present in the embryonic than in the mature brain. We show further that glial cells can incorporate cartilage aggrecan into their PCMs, and that both endogenous and aggrecan supplemented glial PCMs are dependent on hyaluronan. In contrast, primary neurons from newborn (P0) and P1 rat cortex neither express endogenous matrices nor can assemble exogenous hyaluronan/aggrecan aggregates into PCMs. These results suggest that immature neurons may not have the ability to assemble hyaluronan based PCMs, and they raise the possibility that neural proteoglycans associate with neuronal surfaces through a mechanism that may not directly involve hyaluronan. PMID- 9215729 TI - Toxicity of ricin and volkensin, two ribosome-inactivating proteins, to microglia, astrocyte, and neuron cultures. AB - Ricin and volkensin, two potent toxins belonging to the family of ribosome inactivating proteins (RIPs), have been largely exploited in recent years in in vivo experiments of neuronal degeneration consequent to suicide transport or immunolesioning. We have determined both the toxicity of, and the inhibition of, protein synthesis by ricin and volkensin in in vitro cultures enriched in microglial cells, astrocytes, or neurons. In microglial cultures, 50% of toxicity (estimated by LDH released from dead cells) after 24 h exposure to RIPs was obtained with volkensin at 2.2x10(-12) M concentration and 50% of protein synthesis inhibition at 2x10(-14) M concentration. Both values were higher by about one order of magnitude in astrocyte-enriched cultures. Toxicity of, and inhibition of, protein synthesis by, ricin were lower for both cell types by about 1 order of magnitude as compared to volkensin. Cerebellar granule neurons in culture survived remarkably well to 24 h exposure to ricin or volkensin, although their protein synthesis was effectively inhibited by the two toxins with a potency similar to that found for astrocytes. These results demonstrate that glial cells, in particular microglia, are very sensitive to RIPs toxicity and should, therefore, be a primary target of these toxins when injected in vivo. Thus, the damage observed after in vivo experiments could be partly related to diffusion of toxic substances from early-affected glial cells. PMID- 9215730 TI - Loss of inwardly rectifying potassium currents by human retinal glial cells in diseases of the eye. AB - We compared the inward K+ currents of Muller glial cells from healthy and pathologically changed human retinas. To this purpose, the whole-cell voltage clamp technique was performed on noncultured Muller cells acutely isolated from human retinas. Cells originated from retinas of four healthy organ donors and of 24 patients suffering from different vitreoretinal and chorioretinal diseases. Muller cells from organ donors displayed inward K+ currents in the whole-cell mode similar to those found in other species. In contrast, this pattern was clearly changed in the Muller cells from patient retinas. In whole-cell recordings many Muller cells had strongly decreased inward K+ current amplitudes or lost these currents completely. Thus, the mean input resistance of Muller cells from patients was significantly increased to 1,129 +/- 812 M omega, compared to 279 +/- 174 M omega in Muller cells from healthy organ donor retinas. Accordingly, since the membrane potential is mainly determined by the K+ inward conductance in healthy Muller cells, a large amount of Muller cells from patient retinas had a membrane potential which was significantly lower than that of Muller cells from control eyes. The mean membrane potentials were -37 +/- 24 mV and -63 +/- 25 mV for patient and donor Muller cells, respectively. The newly described membrane characteristic changes of Muller cells from patient eyes are assumed to interfere severely with normal retinal function: (1) the retinal K+ homeostasis, which is partly regulated by the Muller cell-mediated spatial buffering, should be disturbed, and (2) the diminished membrane potential should influence voltage-dependent transporter systems of the Muller cells, e.g., the Na(+)-dependent glutamate uptake. PMID- 9215731 TI - Response of Schwann cells to mitogens in vitro is determined by pre-exposure to serum, time in vitro, and developmental age. AB - We compared the mitogenic response of Schwann cells freshly isolated from adult, neonatal, and embryonic nerves, and compared these cells with cells that had been cultured in serum for 5 days. DNA synthesis in response to growth factors was measured using bromodeoxyuridine and immunocytochemistry. Freshly isolated adult Schwann cells were unresponsive to growth factors with or without forskolin to elevate intracellular cAMP levels. After 5 days of culture in serum, or alternatively in defined medium containing fibroblast growth factor 2 plus forskolin, or neu-differentiation factor beta2, adult cells were responsive to mitogens, whereas cells cultured in defined medium alone remained unresponsive. Serum also increased expression of type 1 fibroblast growth factor receptor. Freshly isolated embryonic and neonatal Schwann cells in contrast responded to growth factors even in the absence of forskolin. This responsiveness changed with time in culture. Neonatal cells cultured for 5 days in defined medium in the presence or absence of serum no longer responded to FGF alone, but required forskolin for a mitogenic response. Thus, the response of freshly isolated cells to mitogens is developmentally regulated; extrinsic signals are required to render adult cells responsive to mitogens; and with time in culture, neonatal cells develop a requirement for cAMP elevation for mitogenic response. PMID- 9215732 TI - Effects of tenascin-C in cultured hippocampal astrocytes: NCAM and fibronectin immunoreactivity changes. AB - Tenascin-C is an extracellular matrix glycoprotein with trophic and repulsive properties on neuronal cells, involved in migratory processes of immature neurons. Previous reports demonstrated that this molecule is produced and secreted by astrocytes, in vitro after activation by bFGF or in vivo after CNS lesion. In injured brain the expression of tenascin-C has been correlated with the glial reaction since it was observed in regions suffering a dramatic glial proliferation and hypertrophy. In this report we show that the treatment of cultured hippocampal astrocytes with tenascin-C results in an increased fibronectin and NCAM immunoreactivities. In addition, treated astrocytes form longer extensions than control ones. The number of cells as well as the levels of GFAP mRNA and protein immunoreactivity are not modified after tenascin-C treatment. The present changes may, therefore, be related to the modification of the adhesive properties of astrocytes to the substrate. These observations are compatible with the hypothesis that tenascin-C may contribute to the glial scarring process. PMID- 9215733 TI - Microglia in the pineal gland of the neonatal rat: characterization and effects on pinealocyte neurite length and serotonin content. AB - Microglia in the pineal gland of 1-day-old Sprague-Dawley rats were examined by OX-42 immunocytochemistry and DiI-acetylated-LDL uptake in pineal cell suspension and were found to comprise 3-5% of the total cells in the pineal gland of the neonates. In order to investigate the effects of microglia on pinealocyte structure and function, microglia-depleted and microglia-enriched pineal cell cultures were generated from 1-day-old neonate by fluorescence activated cell sorting (FACS). After 7 days of culture, tissues were processed for either immunocytochemistry for pinealocyte S-antigen and serotonin or high performance liquid chromatography to measure serotonin. Morphometric analysis of immunoreacted cells revealed that pinealocyte neurite length was enhanced in microglia-depleted cultures and was inhibited in a microglia-enriched environment (ANOVA, P < 0.001). Serotonin content of pineal cultures decreased in microglia depleted cultures and was elevated in microglia-enriched cultures (ANOVA, P < 0.001) without any significant change in pinealocyte numbers. These findings are consistent with a working hypothesis that microglia function to mediate neuroendocrine-immune interactions of the gland. PMID- 9215734 TI - Increased potassium, chloride, and taurine conductances in astrocytes during hypoosmotic swelling. AB - Membrane conductances during hypoosmotic swelling were characterized in rat astrocytes in primary tissue culture. Using whole cell patch clamp techniques, mean +/- SEM cell conductance in isoosmotic phosphate-buffered saline (PBS) was 55.6 +/- 5.8 pS/pF. Cell conductance (mean +/- SEM) increased from this initial value to 187 +/- 46%, 561 +/- 188%, and 1216 +/- 376% within 9 min of exposure to 220 mOsm, 190 mOsm, and 145 mOsm PBS, respectively. With each of these hypoosmotic exposures, no change occurred in membrane capacitance. When CsCl replaced KCl in the microelectrode solution, a similar conductance increase was obtained at each osmolality. However, when gluconate salts were used in place of chloride salts in the electrode solution, no significant conductance increase was observed with 190 mOsm PBS. With a KCl microelectrode solution, all conductance increase which occurred in 190 mOsm PBS was inhibited by 200 microM niflumic acid, but not by 5 mM BaCl(2). Both niflumic acid and BaCl(2) inhibited 60-80% of the conductance increase of cells in 145 mOsm PBS. Using a microelectrode solution containing taurine as the major anion, membrane conductance increased 5 fold when cells were placed in 250 mOsm medium. This conductance increase was completely inhibited by 200 microM niflumic acid. Thus, independent chloride and potassium conductances are activated by hypoosmotic swelling of cultured astrocytes while plasma membrane area is unaltered. The chloride conductance pathway is activated at a significantly lower degree of hypoosmotic exposure than that which activates the potassium pathway and may be permeable to anionic taurine. These conductance pathways may mediate diffusive loss of potassium, chloride, and taurine from these cells during volume regulation following hypoosmotic swelling. PMID- 9215735 TI - Expression of neural cell adhesion molecule (NCAM) characterizes a subpopulation of type 1 astrocytes in human optic nerve head. AB - The human optic nerve contains a heterogeneous population of astrocytes. In situ, a specialized subpopulation of astrocytes was distinguished in the adult optic nerve head by expression of neural cell adhesion molecule (NCAM). To further study the biology of astrocytes, we have developed and characterized cells grown from explanted optic nerve heads and myelinated optic nerves as in vitro model systems. Second or third passage cells were processed for immunocytochemistry using antibodies against glial fibrillary acidic protein (GFAP) and cell surface epitopes: CD56/NCAM, HNK-1/NCAM, A2B5, and O4. Synthesis and gene expression of NCAM were characterized by Western blot analysis and RNase protection assay. Cells grown from myelinated optic nerves expressing GFAP, but not NCAM or A2B5, were identified as type 1A astrocytes, and cells expressing GFAP and A2B5, but not NCAM, were identified as type 2 astrocytes. Cells grown from explanted optic nerve head expressing GFAP, NCAM, and O4 were identified as type 1B astrocytes. Expression of NCAM by type 1B astrocytes may provide these cells with adhesion properties that allow them specialized responses in their microenvironment. Astrocytes from the lamina cribrosa may form a functional barrier to prevent myelination of the retina. In glaucoma, these astrocytes may be exposed to stresses due to fluctuation in intraocular pressure and therefore participate in the optic nerve changes associated with glaucomatous optic neuropathy. PMID- 9215736 TI - A simple and reliable method for screening retroviral producer clones without selectable markers. AB - Simplified retroviral vectors that lack dominant selectable markers are being used with increasing frequency. These simplified vectors may offer a number of advantages over selectable marker-containing constructs, including potentially higher titers and less immunogenicity. However, the use of these vectors has been limited by the cumbersome experimental approaches in establishing and characterizing useful producer cell clones. To address this issue, a simple and reliable assay was developed to identify retroviral producer cell lines with or without dominant selectable markers. Producer cells were first generated by standard transfection/transduction and clones isolated by limiting dilution. Supernatant from each clone was then screened by RNA dot blot to identify the best producer clone candidates. The semiquantitative nature of the RNA dot blot assay was validated using a retroviral vector containing neomycin phosphotransferase (neo). Titers obtained by conventional G418-resistant colony forming units/ml (G418(R) cfu/ml) assays strongly correlated with the values by RNA dot blot procedure. RNA dot blot results also correlated well with titers estimated by Southern analysis of HeLa cells transduced with supernatant from each clone. The RNA dot blot technique is a rapid (2 days) and reliable method to screen retroviral producer cells, thereby facilitating the generation and characterization of simplified retroviral producer cell clones. PMID- 9215737 TI - Liver-directed gene transfer in non-human primates. AB - To develop a primate model for liver-directed gene therapy, we studied several gene transfer vehicles and routes in eight rhesus monkeys (Macaca mulatta). For this purpose, we used first-generation, replication-deficient adenoviral vectors carrying the Escherichia coli lacZ gene (Ad.CMVlacZ) or a lacZ-containing plasmid (pCMV beta) with lipofectamine for transfection. The reporter gene construct was infused into either the portal vasculature, common bile duct, or saphenous vein. Adenovirus-mediated gene transfer via the portal vein resulted in expression of lacZ in over 70% of hepatocytes by days 3-7, but was accompanied by acute hepatitis. Adenovirus-mediated gene transfer via the common bile duct resulted in lacZ expression in less than 10% of hepatocytes and was accompanied by portal inflammation. The animals mounted a significant immune response, as demonstrated by adenoviral antigen-induced T-cell proliferation and production of neutralizing anti-adenovirus antibodies and antibodies to E. coli beta-galactosidase (beta Gal). Activation of the immune response was associated with rapid decrease of the reporter gene by days 13-21. Lipofectamine-mediated gene transfer was inefficient, and no lacZ expression in the liver was detected. To limit the host immune response, 4 animals were immunosuppressed by cyclophosphamide/prednisone and then infused with the Ad.CMVlacZ via the portal vein or the saphenous vein. The monkeys showed sustained expression of lacZ for up to 35 days with no evidence of inflammation. The primates transduced via the saphenous vein showed a level of beta-Gal expression in the liver similar to that of the portal vein infused animals. In conclusion, adenovirus-mediated gene transfer to non-human primate livers via the portal vein or saphenous vein is efficient, but it results in transient expression and is accompanied by an immune response to both vector and transgene products and acute hepatitis, whereas lipofectamine-mediated transfer is inefficient. Manipulation of the host immune response may expand potential applications of adenoviral vectors for liver-directed gene transfer. PMID- 9215738 TI - Cytotoxic T lymphocyte responses to proteins encoded by heterologous transgenes transferred in vivo by adenoviral vectors. AB - Although replication-deficient adenovirus (Ad) vectors are efficient vehicles for in vivo gene transfer, persistence of expression of the Ad genome is limited in immunocompetent hosts by cellular immunity directed against the gene product of the vector. While most attention has been focused on cytotoxic T lymphocytes (CTL) directed against the low-level early and late Ad gene expression in the Ad vector-infected target cells, significant cellular immunity is likely also directed against the product of heterologous transgenes. To evaluate this concept, in vivo generation of CTL was evaluated in C57B1/6 and BALB/c mice with Ad vectors expressing a variety of heterologous transgenes, including Escherichia coli chloramphenicol acetyl transferase (CAT), beta-galactosidase (beta-Gal), cytosine deaminase, and human thrombopoietin (hTPO), with an Ad vector expressing no transgene ("null") as a control. Following intravenous administration of Ad vectors, spleen cells were harvested 2 weeks later, stimulated for 5 days with syngeneic cells infected with various Ad vectors, and then evaluated for CTL activity using 51Cr-release from syngeneic Ad vector-infected targets. In all cases, CTL directed against the heterologous transgene products was observed, although there were differences in the amounts of transgene-specific CTL. CTL directed against the transgene were also observed with other routes of administration, including intratracheal, subcutaneous, and intraperitoneal administration. These observations suggest that inclusion of a heterologous transgene in Ad vectors enhances the elimination of vector-infected cells, a circumstance that will be partially circumvented using autologous genes. For some applications, specific immune responses to products of transgenes delivered by Ad vectors might be exploited for therapeutic purposes. PMID- 9215739 TI - In vitro and in vivo secretion of cloned antibodies by genetically modified myogenic cells. AB - In vivo production of recombinant antibodies by engineered cells may have applications for gene therapy of certain cancers and of certain severe viral diseases. It would also permit the development of new animal models of autoimmune diseases and new approaches for in vivo ablation of specific cell types for fundamental purposes. Using gene transfer of an anti-human thyroglobulin monoclonal antibody, we show here that several cell types permitting autologous grafting of genetically engineered cells are efficiently able to secrete antibodies in vitro. Those cells include skin fibroblasts, hepatocytes, and myogenic cells. We also show that the secreted antibodies display an affinity for the antigen close to that of the parental antibody, with, however, slight differences varying according to the cell type. This indicates that the foldings of antigen combining sites of antibodies produced in B cell- and non-B cell contexts are very similar. Finally, we report that, when implanted in the forelimb of a mouse, genetically modified myogenic cells are able to secrete antibodies for at least 4 months. Taken together, our observations point to the notion that genetic modification of patient cells may be used for long-term antibody-based gene therapies. PMID- 9215740 TI - Evaluation of PCR and ELISA assays for screening clinical trial subjects for replication-competent retrovirus. AB - Gene delivery via murine-based recombinant retroviral vectors is currently widely used in gene therapy clinical trials. The vectors are engineered to be replication defective by replacing the structural and nonstructural genes of a cloned infectious retrovirus with a therapeutic gene of interest. The retroviral particles are currently generated in packaging cell lines, which supply all retroviral proteins in trans. Recombination between short homologous regions of the retroviral vector and packaging cell line elements can theoretically generate replication-competent retrovirus (RCR) and hence the Food and Drug Administration (FDA) requires the monitoring of clinical trial subjects for the presence of RCR. Sensitive polymerase chain reaction (PCR) assays have been used for the detection of murine leukemia virus (MLV) nucleotide sequences in peripheral blood mononuclear cells (PBMCs). A novel serological enzyme-linked immunosorbent assay (ELISA) for the detection of anti-MLV specific immunoglobulin (Ig) has been developed to be used as an alternative to the PCR assay. Both assays were used to monitor human immunodeficiency virus (HIV)-positive clinical trial subjects who had received multiple injections of HIV-IT (V), a retroviral vector encoding HIV 1 IIIBenv/rev. Western blot analysis and an in vitro vector neutralization assay were used to characterize further a subset of serum samples tested by ELISA. Results show no evidence of RCR infection in clinical trial subjects. PCR and ELISA assays are discussed in terms of their advantages and limitations as routine screening assays for RCR. The PCR assay is our current choice for monitoring clinical trial subjects receiving direct administration of vector, and the ELISA is our choice for those receiving ex vivo treatment regimens. PMID- 9215741 TI - Nonviral gene delivery to the rat kidney with polyethylenimine. AB - The aim of this study was to establish a nonviral method for gene delivery to the rat kidney. To this purpose, a panel of reagents was tested, including a monocationic lipid, DOTAP, a polycationic lipid, DOGS (or Transfectam), and three different forms of the cationic polymer polyethylenimine (PEI). A comparison among these compounds was performed in vivo, using luciferase as reporter gene. Complexes containing 10 microg of DNA were injected into the left renal artery of rats and allowed to remain in contact with the kidney for 10 min. Forty-eight hours later, luciferase expression levels in kidney extracts were measured. Kidneys injected with DNA complexed to the branched, 25-kD PEI polymer (PEI 25k) yielded activity levels significantly higher than control, sham-operated kidneys (2.7 x 10(4) vs. 5.2 x 10(3) RLU/kidney, respectively), whereas the other transfecting agents did not yield significant activity over controls. PEI 25k was therefore chosen for further optimization of transfection conditions. Dose dependent luciferase expression was shown for 10, 50, and 100 microg of PEI complexed DNA, reaching maximal levels of 2.4 x 10(5) RLU/kidney at 100 microg DNA. The optimal PEI nitrogen/DNA phosphate molar ratio was 10 equivalents. Luciferase activity peaked at 2 days, was still significantly higher than controls at 7 days, and was undetectable at 14 days post-injection. Using beta galactosidase (beta-Gal) as a reporter, transgene expression was localized almost exclusively in proximal tubular cells. PMID- 9215742 TI - Gene transfer using a novel fusion protein, GAL4/invasin. AB - The delivery of DNA to target cells using simple, defined, nonviral systems has become an area of intense interest in gene therapy. We describe here the development and characterization of one such novel system. A recombinant, bifunctional, fusion protein was expressed and purified from Escherichia coli. This protein consists of the DNA-binding domain of the yeast transcription factor GAL4 fused to the cell binding, internalization domain of the Yersinia pseudotuberculosis inv gene product, invasin. This protein, GAL4/Inv, together with poly-L-lysine, formed complexes with a chloramphenicol acetyltransferase (CAT) reporter plasmid that contains eight repeats of the GAL4 consensus recognition sequence. These complexes were shown to transfect target cells in an invasin receptor-dependent manner, resulting in transient CAT expression. A simple, targeted DNA delivery vehicle, as we describe here, represents a viable approach to nonviral gene delivery. PMID- 9215743 TI - Anti-vector immunoglobulin induced by retroviral vectors. AB - Replication-incompetent retroviruses have been employed as gene therapy vectors in experimental settings for more than a decade. More recently, these vectors have been tested in the clinic as immunotherapeutic agents and anticancer agents. One potential problem with the use of such vectors is the possible development of immune responses directed against the vector particles themselves. Here, we examine immunoglobulin (Ig) responses specific for retroviral vectors derived from murine leukemia virus (MLV). Anti-MLV Ig is seen following intramuscular (i.m.) administration of retroviral vectors in mice, and in nonhuman primates; as expected, these responses are dependent upon the vector dose delivered. Furthermore, serum from vector-treated animals is capable of partially neutralizing vector-mediated transduction of target cells in an in vitro assay. Nevertheless, even in the presence of significant levels of anti-vector Ig in vivo, i.m. administration of retroviral vector is still capable of driving both Ig and cytotoxic T lymphocyte (CTL) responses specific for vector-encoded gene products. This work suggests that although retroviral vectors may readily induce immune responses directed against the vector particles themselves, such responses will not significantly affect the efficiency of these vectors in an immunotherapeutic protocol. PMID- 9215744 TI - Immune responses to reporter proteins and high viral dose limit duration of expression with adenoviral vectors: comparison of E2a wild type and E2a deleted vectors. AB - Experiments designed to evaluate the effect of deletion of E2a on duration of expression using adenoviral vectors led to a series of observations regarding host responses to adenoviral vectors and reporter proteins. In studies using human alpha1-antitrypsin (hAAT) as a reporter gene, we found that the duration of expression is very brief for C3H/J and CBA/J mice but is prolonged for C57BL/6J mice, that disappearance of hAAT from the blood is correlated with the appearance of antibodies, and that immunization against hAAT can prevent appearance of the protein in the blood after administration of an adenoviral vector. Deletion of E2a in hAAT vectors did not prolong expession in C3H/J or CBA/J mice and did not shorten duration of expression in C57BL/6J mice. Using similar vectors expressing Escherichia coli beta-galactosidase (beta-Gal) in immunocompetent mice, short duration of expression with a beta-Gal reporter was remarkably different from the long expression with an identical vector expressing hAAT in C57BL/6J. In the case of vectors expressing hAAT, adenoviral sequences persisted in the liver, and inflammatory responses were minimal compared to vectors expressing beta-Gal, where adenoviral sequences disappeared from the liver concomitant with a prominent inflammatory response. The duration of expression of beta-Gal in hepatocytes was increased in transgenic mice expressing the reporter in keratinocytes, indicating that host immune responses to the reporter can limit duration of expression. Dosage studies indicated that persistence of expression of hAAT can be markedly decreased by administration of high doses of vector in a manner consistent with a nonimmune-mediated toxicity following injection. These experiments indicate that host responses to reporter genes rather than host responses to adenoviral proteins can be the primary determinant of duration of expression under many experimental conditions. PMID- 9215746 TI - Hematologically important mutations: Gaucher disease. PMID- 9215745 TI - A pilot study of idiotypic vaccination for follicular B-cell lymphoma using a genetic approach. CRC NO: 92/33. Protocol NO: PH1/027. PMID- 9215747 TI - Modulatory subdomains of the HS2 enhancer differentially regulate enhancer activity in erythroid cells at different developmental stages. AB - The HS2 enhancer in the locus control region of human beta-like globin genes displays developmental-stage-independent enhancer function. The mechanism by which it regulates the transcription of the globin genes in erythroid cells throughout development is not fully understood. In this paper we dissect the HS2 enhancer into an enhancer core and five modulatory subdomains M1 to M5. The enhancer core possesses developmental-stage-independent enhancer activity. The modulatory subdomains by themselves do not possess such enhancer activity, but they apparently respond to environmental signals and modulate enhancer core activity in a developmental-stage specific manner. M1 located 5' of the core strongly stimulates core activity in K562 cells at the embryonic stage. M2 and M3 located 3' of the core strongly stimulate core activity in MEL cells at the adult stage. Moreover, M3 suppresses core activity at the embryonic stage and exhibits an adult-stage-selector activity. These findings indicate that the apparent developmental-stage-independence of the HS2 enhancer is a result of multiple interactions between the core and the modulatory subdomains located both near and far from the core. PMID- 9215748 TI - Expression of transcription factors during sodium phenylacetate induced erythroid differentiation in K562 cells. AB - During 15 days of treatment of K562 cells with sodium phenylacetate, we observed an increase in the cellular hemoglobin concentration with a similar increase in the expression of gamma-globin mRNA. Morphological studies demonstrated characteristic features of erythroid differentiation and maturation. At the same time there was no change in the level of expression of the cell surface antigenes CD33, CD34, CD45, CD71 and glycophorin A. Likewise, the level of expression of the erythroid transcription factors GATA-1, GATA-2, NF-E2, SCL and RBTN2, all expressed in untreated K562 cells, did not increase during sodium phenylacetate induced erythroid differentiation. The expression of the nuclear factors Evi-1 and c-myb, known to inhibit erythroid differentiation, did not decrease. We conclude that sodium phenylacetate treatment of K562 cells increases gamma-globin mRNA and induces cell maturation as judged by morphology without affecting the expression of the erythroid transcription factors, some of which are known to be involved in the regulation of beta-like globin genes. PMID- 9215749 TI - Hematologically important mutations: Glanzmann thrombasthenia. PMID- 9215750 TI - A hemophilia model in zebrafish: analysis of hemostasis. AB - We have developed artificial hemophilia in zebrafish by treating them with copper and measured their clotting function by a newly developed sensitive clotting time assay. The clotting function can be detected rapidly and reliably in 30 hr larvae and in adult fish by measuring the blood clotting time. We have used this assay to screen wild type zebrafish and identified fish with prolonged clotting time. This verifies the usefulness of this assay in future screening for recessive hemostasis defects generated by chemical and radiation mutagenesis methods. PMID- 9215751 TI - Red cell-mediated therapy: opportunities and challenges. AB - Red blood cells have unique properties that offer attractive therapeutic possibilities. It is proposed that by covalently linking drug pharmacophores to red cell surface proteins, significant enhancement of pharmacokinetics can be achieved. Specific anchoring to erythrocytes can be accomplished in principle, using unique, affinity labeling, combinatorial libraries targeting the surface protein, glycophorin A. The anticipated advantages of red cell-anchored drugs over free drugs, include extended half-lives, controlled volumes of distribution, and multivalent interactions. PMID- 9215752 TI - Cationic phosphonolipids as non viral vectors for DNA transfection in hematopoietic cell lines and CD34+ cells. AB - The ability to transfer genes into a hematopoietic stem cell and to achieve regulation of their expression in lymphoid or myeloid lineages should open many new therapeutic opportunities. Besides gene transfer mediated by virus vectors like retrovirus or adenovirus, non viral systems have the theoretical advantage of being safe and easy to manage. We developed a new family of cationic lipids called phosphonolipids, synthesized 24 new molecules, and then in a first step we tested their potential to transfer genes in human hematopoietic cell lines (K562 and TF1). A LacZ plasmid under the control of a strong viral promoter was used as a reporter gene and a FACS-Gal assay and a quantitative test CPRG assay evaluated the beta gal expression. The targeted cells were analyzed 48 hours after transfection. The present work shows that seven novel molecules display a high transfer efficiency. One of them is nine-fold more efficient than the commercially available cationic lipids. The results obtained ex vivo on CD34 cells with the FACS-Gal assay show that at day 10 after transfection, 45 percent of cells are expressing gal. PMID- 9215753 TI - Molecular genetics of glucose-6-phosphate dehydrogenase deficiency in Mexico. AB - Several studies carried out between 1965 and 1985 showed that G-6-PD deficiency in Mexico is heterogeneous at the biochemical level and that the G-6-PD A- phenotype is relatively common. We have now investigated the molecular basis of G 6-PD deficiency in Mexico. Up-to-date 60 chromosomes with G6PD mutations have been studied, 16 in previous studies and 44 in the present work. Molecular analysis of DNA from G-6-PD deficient Mexican mestizos and their relatives show that G-6-PD A- genotypes are relatively common but also that in Mexico G-6-PD deficiency is heterogeneous at the DNA level. Thus, five different genotypes have been observed: G-6-PD A-(202A/376G) (41 chromosomes), G-6-PD A-(376G/968C) (14 chromosomes), G-6-PD Seattle844C (3 chromosomes), G-6-PD "Mexico City"680A (1 chromosome) and G-6-PD Guadalajara1159T (1 chromosome). The G-6-PD A-(202A/376G), G-6-PD A-(376G/968C) and G-6-PD Seattle844C mutations in Mexico are on the same Pvu II/ Pst I/ 1311 / Nla III haplotypes as found in individuals from Africa, Spain and the Canary Islands. Consequently, these mutations were probably imported to Mexico through African slaves and/or the Spanish immigrants during and after the colonization. PMID- 9215754 TI - HLA-H mutations in the Ashkenazi Jewish population. AB - Hereditary hemochromatosis is a common disorder in people of European origin. The HLA-H gene has been found to have two mutations that apparently cause hemochromatosis. The principal mutation, 845G-->A (C282Y), is believed to have arisen relatively recently in the Celtic population. To determine the incidence of this mutation and the other hemochromatosis-associated mutation, 187C-->G (H63D), among Ashkenazi Jews, a people who are believed to have arrived in Europe in about the 8th Century A.D., we have examined the DNA from 381 unrelated Jewish subjects and 206 non-Jewish white controls. The gene frequency for the 845G-->A mutation among Jewish subjects was only 0.013 compared with a frequency of 0.070 among controls, a difference that is significant at the 0.00001 level. The phenotypically milder nt 187C-->G mutation had a frequency of 0.155 in the non Jewish population and 0.097 in the Jewish population, a difference that was also statistically significant at the <0.01 level. PMID- 9215755 TI - Evidence for an uncommon microsatellite instability on mouse chromosomes 2 and 4 and its possible role in radiation leukemogenesis. AB - Although microsatellite instability (MSI), usually detected by DNA length polymorphisms, has been implicated in the induction of solid tumors in both humans and animals, its role in leukemogenesis is unclear. The goal of this study was to investigate whether there is an association between MSI and radiation leukemogenesis in CBA/Ca mice. Microsatellite lengths at 55 loci, mapped to eight different mouse chromosomes, were examined in two groups of DNA samples: 1) 10 normal DNA samples collected from the bone marrow cells of control male CBA/Ca mice, and 2) 17 DNA samples isolated from the spleens of mice that developed myeloid leukemia (ML) after exposure to neutrons, or X rays, or gamma rays. Microsatellite markers were amplified using the non-radioisotopic multiplex touchdown PCR protocols developed in our laboratory, and the sizes of amplicons were examined on 6% non-denaturing polyacrylamide gels. Although no correlation between microsatellite length polymorphisms and radiation leukemogenesis was observed at the 55 CBA/Ca mouse loci tested in this study, an uncommon MSI, manifested as the absence of DNA bands after PCR amplification at 2 loci (D2MIT140 and D4MIT104), was observed in both control and ML samples. However, the frequency of ML samples showing this type of MSI is statistically significant (p<0.05). Although there is no direct evidence that this type of MSI predisposes mice to the development of leukemia, the results suggests that genes flanking the D2MIT140 and D4MIT104 are susceptible to spontaneous mutation and perhaps to damage caused by ionizing radiation. PMID- 9215756 TI - T-cell receptor beta chain variability in bone marrow and peripheral blood in severe acquired aplastic anemia. AB - Aplastic anemia (AA) is characterized by multilineage bone marrow failure of unknown etiology. In order to assess the role of immune-mediated mechanisms in hematopoietic suppression, we examined the diversity of T lymphocyte repertoire in terms of variable (V) gene segment usage of the T cell receptor (TCR) beta chain in bone marrow and peripheral blood of six patients with severe untreated AA. Expression of transcripts encoding Vbeta1-Vbeta24 subfamilies was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). The results revealed that T lymphocytes in AA utilize highly diverse segments of the beta chain loci. Over the heterogenous Vbeta expression background, transcripts encoding Vbeta3, Vbeta20, Vbeta21, and Vbeta22 subfamilies were enhanced by at least threefold in 5 of 6 patients as compared to normal samples, but a different transcript species was over expressed in each patient. To evaluate clonality of T cells, size diversity within the complementarity determining region 3 (CDR3) and usage of TCRbeta joining (J) gene segments were analyzed in PCR products specific for each of the 24 Vbeta subfamilies. We found that the majority of transcripts display normal CDR3 size patterns, as is characteristic of polyclonal populations. Nevertheless, one or two predominating junctional rearrangements were observed in each patient. They were identified in Vbeta5, Vbeta7, Vbeta8, Vbeta13, Vbeta15, Vbeta16, and Vbeta23 transcripts, which differed from patient to patient and did not correspond to transcripts with an abnormally high expression level. Our results demonstrate that T cell repertoire in AA is random with respect to the TCR beta chain. Unique rearrangements detected in the CDR3 region are suggestive of a limited process of an antigen-driven (oligo)clonal T cell expansion which may take place over the overwhelmingly polyclonal repertoire of T lymphocytes at the onset of severe AA. PMID- 9215757 TI - Crystal structure of NMC-4 fab anti-von Willebrand factor A1 domain. AB - We have solved the crystal structure of the Fab fragment of NMC-4, a mouse monoclonal antibody that binds to the A1 domain of von Willebrand factor (vWF). Two Asp and three Tyr residues in the complementarity determining regions 1 and 3 of the heavy chain exhibited a spatial orientation suggestive of a dominant role in establishing contact with the antigen. A cluster of Asp and Tyr residues occurs also in a region of the platelet glycoprotein (GP) Ib alpha amino terminal domain known to be critically involved in vWF binding. Thus, the structural information obtained with NMC-4 may prove relevant to understand the stereochemical bases of the GP Ib alpha-vWF interaction essential for thrombus formation at sites of vascular lesion. PMID- 9215758 TI - Genetic and clinical description of hemochromatosis probands and heterozygotes: evidence that multiple genes linked to the major histocompatibility complex are responsible for hemochromatosis. AB - We evaluated Alabama hemochromatosis probands (n = 74) and normal control subjects (n = 142) for expression of the hemochromatosis-associated mutations nt 845G-->A (845A; Cys282Tyr) and nt 187C-->G (His63Asp) in a gene linked to the major histocompatibility complex (MHC). We also tabulated parameters of iron metabolism and iron overload in probands and in obligate heterozygote family members of homozygous Cys282Tyr probands. Among probands, 59.4% were Cys282Tyr homozygotes and 20.3% were heterozygotes; 20.3% did not express this mutation. In normal control subjects, 14.7% were heterozygous for the Cys282Tyr mutation; one normal control subject was homozygous for the Cys282Tyr mutation. None (0 of 44) of our Cys282Tyr-homozygous hemochromatosis probands had the His63Asp mutation. Of the Cys282Tyr-heterozygous and -negative probands, the His63Asp mutation occurred in 26.7% (4/15) and 53.3% (8/15), respectively. In normal control subjects, 23.2% were heterozygous for the His63Asp mutation; 2.8% were homozygous. Induction phlebotomy requirements and other manifestations of iron overload were significantly greater in Cys282Tyr homozygotes than among other probands. Cys282Tyr-heterozygous probands had significantly higher values of serum iron parameters than did obligate Cys282Tyr heterozygotes whose values were, on the average, normal. Co-expression of HLA-A3, HLA-B7, and D6S105(8) was significantly more frequent in all subgroups of probands stratified by Cys282Tyr expression than in normal control subjects. These results demonstrate that the severity of iron overload in hemochromatosis is affected significantly by genetic factors. Further, our findings support the hypothesis that one or more MHC-linked genes other than that corresponding to the Cys282Tyr and His63Asp mutations contributes to increased iron absorption and iron overload in hemochromatosis probands. PMID- 9215759 TI - Using 125I-albumin exclusively for blood volume examinations. PMID- 9215760 TI - Sex chromosome markers: characterization using fluorescence in situ hybridization and review of the literature. AB - Fluorescence in situ hybridization (FISH) using biotin labeled X- and Y chromosome DNA probes was utilized in the analysis of 23 sex chromosome-derived markers. Specimens were obtained through prenatal diagnosis, because of a presumptive diagnosis of Ullrich-Turner syndrome, mental retardation, and minor anomalies or ambiguous genitalia; three were spontaneous abortuses. Twelve markers were derived from the X chromosome and eleven from the Y chromosome; this demonstrates successfully the value and necessity of FISH utilizing DNA probes in the identification of sex chromosome markers. Both fresh and older slides, some of which had been previously G-banded, were used in these determinations. We have also reviewed the literature on sex chromosome markers identified using FISH. PMID- 9215761 TI - The spectrum of congenital anomalies of the VATER association: an international study. AB - The spectrum of the VATER association has been debated ever since its description more than two decades ago. To assess the spectrum of congenital anomalies associated with VATER while minimizing the distortions due to small samples and referral patterns typical of clinical series, we studied infants with VATER association reported to the combined registry of infants with multiple congenital anomalies from 17 birth defects registries worldwide that are part of the International Clearinghouse for Birth Defects Monitoring Systems (ICB-DMS). Among approximately 10 million infants born from 1983 through 1991, the ICB-DMS registered 2,295 infants with 3 or more of 25 unrelated major congenital anomalies of unknown cause. Of these infants, 286 had the VATER association, defined as at least three of the five VATER anomalies (vertebral defects, anal atresia, esophageal atresia, renal defects, and radial-ray limb deficiency), when we expected 219 (P<0.001). Of these 286 infants, 51 had at least four VATER anomalies, and 8 had all five anomalies. We found that preaxial but not other limb anomalies were significantly associated with any combination of the four nonlimb VATER anomalies (P<0.001). Of the 286 infants with VATER association, 214 (74.8%) had additional defects. Genital defects, cardiovascular anomalies, and small intestinal atresias were positively associated with VATER association (P<0.001). Infants with VATER association that included both renal anomalies and anorectal atresia were significantly more likely to have genital defects. Finally, a subset of infants with VATER association also had defects described in other associations, including diaphragmatic defects, oral clefts, bladder exstrophy, omphalocele, and neural tube defects. These results offer evidence for the specificity of the VATER association, suggest the existence of distinct subsets within the association, and raise the question of a common pathway for patterns of VATER and other types of defects in at least a subset of infants with multiple congenital anomalies. PMID- 9215762 TI - Gaucher disease: enzyme therapy in the acute neuronopathic variant. AB - The responses to regular intravenous enzyme infusions were compared in two sibs with Gaucher disease type 2, the acute neuronopathic variant. Enzyme administration was begun at 7 months in patient 1 who had severe progressive visceral and neuronopathic disease. No significant effect of enzyme infusions was noted. Death occurred at 9 months. Patient 2 was prenatally diagnosed and enzyme infusions were initiated at age 4 days. Overall development progressed at a rate similar to her unaffected full sib until her death at 15.1 months. Slowly progressive esotropia, ocular paresis and dysphagia began at 8 months as did infiltrative pulmonary disease. Comparison of these clinical courses show significant visceral and neurologic effects of anticipatory enzyme therapy, but with unaltered outcome, for Gaucher disease type 2. PMID- 9215763 TI - Developmental tasks of childhood and adolescence: implications for genetic testing. AB - Many reports have recently recommended a careful weighing of the potential benefits and harms of genetic testing (carrier or predisposition) of children and adolescents [Andrews et al., Washington DC: National Academy Press, 1994; Wertz et al., JAMA, 272:875-881, 1994; Clinical Genetics Society (UK), J Med Genet, 31:785-797, 1994; ASHJ/ACMG, Am J Hum Genet, 57:1233-1241, 1995]. Despite this, youngsters are currently being tested for late-onset disorders as well as for carrier status [Reilly and Wertz, Am J Hum Genet, 57:A57, 1995]. Many children to be tested will be those in at-risk families, who may already have experienced the chronic illness or death of a close relative. Thus, reactions to testing will be influenced by prior family experiences. Emotional reactions to testing will be determined by both the child's cognitive and psychosocial development. Testing of adolescents may alter the achievement of developmental tasks, including seeking freedom from parental figures, establishment of personal identity, handling of sexual energies, and remodeling of former idealizations of self and others. There are many potential dilemmas in deciding whether to test a child or adolescent for genetic status. If parents choose not to test, the risk is for later difficulty integrating such information into the self concept. If parents test and do not tell results, the risk is for creating a climate of family secrecy. If parents test and tell results, the risk is robbing the child of the autonomy of his or her own later decision. Perhaps the question of whether to test is not the real question. More than genetic testing, genetic counseling is of crucial importance in thoughtful decisions concerning whether to test an individual child or adolescent. A more important question may be how to provide unaffected children in at-risk families with appropriate counseling. Provision of psychosocial support to at-risk families will enable the child to encounter genetic testing, if necessary, supported with the best possible resources. PMID- 9215765 TI - Genetic disorders among Palestinian Arabs. 2. Hydrocephalus and neural tube defects. AB - Congenital hydrocephalus and/or open neural tube defect was present in at least one individual of 98 families out of the 2,000 Palestinian Arabic families who have visited the Genetic clinic at the Hadassah Medical Center. In 22 families the brain malformation was part of a syndrome: Meckel syndrome in 10, Warburg syndrome in another 5, Carpenter in one, and undiagnosed in 6 families. In 76 of the families the neural tube defect and/or the hydrocephalus were non-syndromal. It seems that most of the cases of isolated non-syndromal hydrocephalus represented autosomal recessive traits and that an abnormal allele is common among Palestinian Muslim Arabs. PMID- 9215764 TI - Population study of congenital hypothyroidism and associated birth defects, Atlanta, 1979-1992. AB - Very little data are available from population-based studies on congenital hypothyroidism (CH) epidemiology and patterns of associated birth defects. By linking data from two population-based registries, we describe the epidemiology of CH and associated defects in Atlanta from 1979-1992. Cases included all infants with CH born from 1979-1992 to mothers residing in the metropolitan Atlanta area at the time of birth. We ascertained CH cases by reviewing newborn screening records and records of the Metropolitan Atlanta Congenital Defects Program (MACDP), a population-based registry of all serious birth defects diagnosed during a child's first year of life. We linked CH cases with MACDP records to ascertain the presence of serious birth defects among infants with CH. Of 97 infants identified with CH through newborn screening and/or MACDP (1:5,000 live births), 87 had primary CH and 10 had secondary. The rate of primary CH was higher among non-hispanic whites than among blacks (1:4,400 vs. 1:10,000) and among females compared with males (1:4,000 vs. 1:7,700). Among infants with primary CH, 77 had isolated CH, 3 had Down syndrome, and 7 had unrelated major structural defects. Based on Atlanta population rates of Down syndrome and major structural anomalies, we infer i) infants with Down syndrome have a 35-fold increased risk for primary CH compared with infants in the general population (P < .0001); ii) infants with primary CH have a 2.2-fold increased risk for major structural anomalies (P < .05). Because this is the first population study of CH in the United States in which data from two population-based registries were linked, the epidemiologic patterns and associated defects are more representative than those found in studies based on newborn screening records only. PMID- 9215766 TI - Evidence for genetic anticipation in the oculodentodigital syndrome. AB - Oculodentodigital syndrome (O.D.) is an autosomal dominant disorder comprising facial anomalies, syndactyly, microcorneae, dental enamel hypoplasia, and leukodystrophy. We describe a four generation family with O.D. in which anomalies such as syndactyly appear congenitally, whereas neurological (i.e., leukodystrophic) signs and symptoms tend to be expressed in a more severe form and/or at an earlier age of onset in successive generations of the kindred. This pattern of phenotypic expression is consistent with the phenomenon of genetic anticipation, and we suggest that O.D. may be a trinucleotide repeat disorder. PMID- 9215767 TI - Paternal risk factors for isolated membranous ventricular septal defects. AB - A case-control study using data from the Baltimore-Washington Infant Study (BWIS) examined possible paternal risk factors in the etiology of isolated membranous ventricular septal defects (VSD). There were 641 total VSD case infants and 3,549 randomly selected control infants ascertained between 1981 and 1989. Isolated membranous VSDs were identified in 499 cases. Socio-demographic factors (such as parental age and race), social habits, and medical conditions were analyzed by multiple logistic regression in order to estimate adjusted odds ratios (OR) and 95% confidence intervals (CI). Paternal age was not found to be a risk factor per se, but small positive associations were found for some social habits and maternal factors. Significant associations were found for paternal marijuana use (OR 1.36, 95% CI 1.05-1.76), African-American race of the infant (OR 1.34, 95% CI 1.09-1.65), and for cocaine use among older fathers (OR 3.92, 95% CI 1.30-11.86). These associations support a multifactorial etiologic hypothesis for isolated membranous VSDs and point to some interesting parental behavioral and medical considerations which may contribute to risk for this common birth defect. PMID- 9215768 TI - Delimiting the Wolf-Hirschhorn syndrome critical region to 750 kilobase pairs. AB - Wolf-Hirschhorn syndrome (WHS) is a multiple anomaly condition characterized by mental and developmental defects, resulting from the absence of the distal segment of one chromosome 4 short arm (4p16.3). Owing to the complex and variable expression of this disorder, it is thought that the WHS is a contiguous gene syndrome with an undefined number of genes contributing to the phenotype. The 2.2 Mbp genomic segment previously defined as the critical region by the analyses of patients with terminal or interstitial deletions is extremely gene dense and an intensive investigation of the developmental role of all the genes contained within it would be daunting and expensive. Further refinement in the definition of the critical region would be valuable but depends on available patient material and accurate clinical evaluation. In this study, we have utilized fluorescence in situ hybridization to further characterize a WHS patient previously demonstrated to have an interstitial deletion and demonstrate that the distal breakpoint occurs between the loci FGFR3 and D4S168. This reduces the critical region for this syndrome to less than 750 kbp. This has the effect of eliminating several genes previously proposed as contributing to this syndrome and allows further research to focus on a more restricted region of the genome and a limited set of genes for their role in the WHS syndrome. PMID- 9215769 TI - Infantile spasms in two children with Williams syndrome. AB - We describe two children with Williams syndrome and infantile spasms. The diagnosis of Williams syndrome was confirmed by documentation of a deletion of the elastin gene/Williams syndrome region at 7q11.23. The diagnosis of infantile spasms was confirmed through the presence of interictal hypsarrhythmia. This represents one of the first reports of infantile spasms in the Williams syndrome. PMID- 9215771 TI - BRCA1 in the family: a case description of the psychological implications. AB - Our experience with the first family in the Netherlands for whom predictive DNA testing for Hereditary Breast and Ovarian Cancer (HBOC) became an option is described. This serves to illustrate the complex emotional impact on a family as a whole, and upon the members separately, of becoming aware that breast and ovarian cancer is hereditary, and the implications of undergoing predictive testing. All family members received genetic counseling and were offered pre- and post-test psychological follow-up. We observed two important roles within the family. One member became "the messenger of the news" informing the relatives of the hereditary character of cancer in the family. Another was "the first utilizer" of the new options; namely, the predictive DNA-test and preventive surgery. This first utilizer became the example to the rest of the family. Decisions made about preventive treatment (prophylactic ovariectomy and/or mastectomy) were based on the experiences within the family, whether one identified with an affected family member with breast or with ovarian cancer. The actions and reactions perceived were illustrative of what kind of support provisions should be provided in addition to the genetic and oncological counseling for HBOC. Moreover HBOC should be considered both as an individual and a family problem and be treated as such in genetic counseling. PMID- 9215770 TI - Hypopigmentation in the Prader-Willi syndrome correlates with P gene deletion but not with haplotype of the hemizygous P allele. AB - The Prader-Willi syndrome (PWS) usually results from a paternal deletion of 15q11 q13 or maternal disomy for chromosome 15. Reduced pigmentation of skin, hair, and eyes is common in PWS and was suggested previously to be associated with the 15q11-q13 deletion. The P gene, located in this same region, is associated with OCA2, an autosomal recessive disorder that is the most frequent form of tyrosinase-positive oculocutaneous albinism. We studied 28 individuals with PWS and found that hemizygosity for the P gene was significantly correlated with the occurrence of hypopigmentation among PWS patients. However, we found little or no relationship between the occurrence of hypopigmentation and the polymorphism haplotype of the intact P allele. Thus, our results indicate that hypopigmentation is likely the result of deletion of the P gene in the context of PWS but do not support the linked hypothesis that hypopigmentation results from hemizygosity for variant P alleles with reduced function. PMID- 9215772 TI - Noncompaction of the ventricular myocardium in Melnick-Needles syndrome. AB - A 4-year-old asymptomatic boy was referred for evaluation after being diagnosed with Melnick-Needles syndrome. Echocardiography demonstrated noncompaction of the left ventricular myocardium as an isolated cardiac finding. Though other structural cardiac defects were reported previously in this syndrome, this is the first report of this rare cardiac anomaly in this likewise rare syndrome. We postulate that this occurrence represents a primary disorder of early fetal development. Patients with Melnick-Needles syndrome should be evaluated for noncompaction of the ventricular myocardium and its potential cardiovascular dysfunction. PMID- 9215773 TI - Autopsy, radiographic, and prenatal ultrasonographic examination of a stillborn fetus with femoral facial syndrome. AB - Femoral facial syndrome (FFS) is comprised of cleft palate, micrognathia, short or absent femora, and vertebral and genitourinary malformations. We report on a stillborn fetus with FFS delivered to a mother with gestational diabetes. Prenatal ultrasound examination showed abnormalities at 21 weeks of gestation; prior ultrasound findings were interpreted as normal. Long bone histology showed disorganization of the growth plate with a relative decrease in cartilaginous matrix and vacuolization and binucleation of the chondrocytes. PMID- 9215774 TI - The gene for autosomal dominant hidrotic ectodermal dysplasia (Clouston syndrome) in a large Indian family maps to the 13q11-q12.1 pericentromeric region. AB - Hidrotic ectodermal dysplasia (HED), Clouston syndrome (MIM No. 129500), is an autosomal dominant disorder affecting the skin and its derivatives. It is characterized by alopecia, dysplastic nails in hands and feet, and hyperkeratosis of the palms and soles. We have studied a large Indian pedigree (UR005), from Gujarat region, consisting of a total 127 individuals including 41 affected (12 males and 29 females). The phenotype in this family ranged from atrichosis to hypotrichosis, sparsity or absence of eyebrows, and thickening of palms and soles. In order to map the disease locus by linkage analysis, DNA polymorphisms were used in DNAs from 23 affected and 8 normal individuals. While genotyping was in progress, Kibar et al. [1996] reported mapping of the locus of a similar disease in French-Canadian families to 13q around marker D13S141. We then utilized markers on 13q to genotype the members of the Indian family. Linkage with 13q11-12.1 markers was confirmed with a maximum lod score of 3.27 (theta=0.00) with locus D13S1316. Multipoint linkage analysis yielded a lod score of 5.04 at theta=0.00 with D13S1316; haplotype analysis indicated that the gene for the Clouston syndrome in this family is localized proximal to D13S292. These data suggest that the gene for the Clouston syndrome in this Indian pedigree is probably the same as that described in the French Canadian families. The combination of data from all available families linked to 13q11-12.1 will make it possible to narrow the critical region and facilitate the positional cloning of the elusive gene. PMID- 9215775 TI - Craniotubular dysplasia with severe postnatal growth retardation, mental retardation, ectodermal dysplasia, and loose skin: Lenz-Majewski-like syndrome. AB - The heterogeneous group of craniotubular dysplasias is characterized by modeling errors of the craniofacial and tubular bones. Some conditions in this category cause not only skeletal abnormalities but also a variety of mesoectodermal dysplasias, as exemplified in Lenz-Majewski syndrome (MIM 151050), which comprises craniodiaphyseal dysplasia, failure to thrive, mental retardation, proximal symphalangism, enamel hypoplasia, and loose skin. We report on a boy with a hitherto unknown multisystem disorder, including skeletal changes that were regarded as a form of craniotubular dysplasia. The patient had a large head, exophthalmos, a broad nasal root, anteverted nostrils, large auricles, thick lips, micrognathia, severe postnatal growth retardation with emaciation, severe mental retardation, sparse hair growth, enamel hypoplasia, and thin, loose skin with hyperlaxity. Skeletal changes consisted of thickened calvaria, sclerosis of the skull base and facial bones, thick ribs, and metaphyseal undermodeling of the tubular bones. In addition, generalized osteopenia was evident. The present disorder overlaps phenotypically with Lenz-Majewski syndrome; nevertheless, the absence of diaphyseal hyperostosis and proximal symphalangism in the present patient was not consistent with Lenz-Majewski syndrome. PMID- 9215776 TI - Multivitamin supplementation and multiple births. AB - It is well established that maternal multivitamin supplementation reduces the risk of neural tube defects and evidence suggests that it may be associated with other reproductive outcomes. The present study was prompted by a report from a randomized trial in Hungary which showed a 40% increase in multiple births among periconceptional vitamin users. Retrospectively collected data on multivitamin supplementation were obtained on multiple and singleton births from three separate studies: Atlanta Birth Defects Case-Control Study (ABDCCS) malformed and nonmalformed infants born 1968-1980, California Birth Defects Monitoring Program (CBDMP) malformed and nonmalformed infants born 1987-1989, and Boston University Slone Epidemiology Unit Birth Defects Study (SEU-BDS) malformed infants born 1987 1994. Supplementation was divided into three mutually exclusive categories based on timing: "periconceptional" use--before through at least the third month after conception; "early" use--beginning in the first month and continuing through at least the third month after conception; and "later" use--beginning in the second or third month after conception. For periconceptional use, four of five datasets showed a 30 to 60% greater prevalence of supplementation among mothers of multiple births. In contrast, this pattern was not evident for "early" and "later" use. Overall, the study findings are tentative, due to a lack of consistency across all five datasets and they should not alter recent recommendations related to folate supplementation for the prevention of neural tube defects. PMID- 9215777 TI - Umbilical cord agenesis in limb body wall defect. AB - The term "Limb Body Wall Defect" (LBWD) refers to a variable group of congenital defects having in common abdomino- or thoraco-schisis and limb deficiency. Three general pathogenic mechanisms have been proposed for this disorder: amnion rupture, vascular disruption, and embryonic malformation. We hypothesize that there are subsets of "Limb Body Wall Defect," which have similar structural abnormalities and a common pathogenesis. We report on five cases of LBWD that were selected by using more restrictive criteria. An infant or fetus was included if it had abdominoschisis with a broad attachment of skin to amnion at the site of the abdominal wall defect, limb defects, and umbilical cord agenesis. Autopsy detected additional common structural defects. All had evisceration of the gastrointestinal structures into the extra-embryonic coelomic space with structural abnormalities of the intestines. All had scoliosis, thoracic deformities, pulmonary hypoplasia, and structural abnormalities of the cloaca and urogenital ridge. Four of five had meningomyelocele. Three had exstrophy of the cloaca. The four females all had ovarian agenesis and incomplete Mullerian fusion. None had normal development of the external genitalia. We propose that the pathogenesis was a primary malformation of body wall closure, with abnormal fusion of the amnion, which had occurred in the first month of development. PMID- 9215778 TI - Delayed diagnosis in patients with Prader-Willi syndrome due to maternal uniparental disomy 15. AB - Prader-Willi syndrome (PWS) results from absence of the normally active paternally inherited genes on proximal 15q, due to del(15)(q11q13) or by maternal uniparental disomy (UPD) 15 in most cases. In addition to a higher frequency of hypopigmentation among deletion patients, minor phenotypic differences between deletion and UPD patients have recently been reported, including lower birth weight in the deletion group, shorter birth length in males with UPD, and shorter course of gavage feeding and later onset of hyperphagia in females with UPD. We previously reported that those with UPD had a less "typical" facial appearance, and they less often had skin picking, skill with puzzles, and high pain threshold. There were no children younger than 3.5 years of age in the UPD group, in contrast to several of them in the deletion group, suggesting a possible diagnostic delay in the UPD group. To assess this possibility and seek reasons for it, we reviewed the charts of 60 PWS patients with complete molecular testing. Mean age at diagnosis of patients with UPD was significantly higher than in the deletion group. Mean percentiles of birth weights and lengths of patients with UPD were significantly lower than in those with deletion. Mean duration of gestation, mean duration of gavage feeding, and mean age at onset of hyperphagia did not differ significantly between groups. Delay in the diagnosis of patients with UPD, which may influence the management and impact of the disorder, might be explained by a lower frequency of typical facial anomalies in this group. PMID- 9215779 TI - Cerebral infarction in Noonan syndrome. AB - We report on an infant with severe Noonan syndrome, chylothoraces, and hepatosplenomegaly who suffered two episodes of cerebral infarction before age 6 months. No underlying cause for these events was found. The presentation is discussed in relationship to other reports of stroke in Noonan syndrome which have previously been associated with underlying vascular malformations. PMID- 9215780 TI - Incidence of aneuploid spermatozoa among infertile men studied by multicolor fluorescence in situ hybridization. AB - We studied by fluorescence in situ hybridization the frequency of aneuploidy in spermatozoa of 12 infertile men: 8 with normal or nearly normal semen analysis values and 4 with oligo-astheno-teratozoospermia. The control group consisted of 18 normal healthy fertile men. Probes for chromosome 1 and 7 were used and 10,000 spermatozoa per individual were scored. The hybridization efficiency was good (higher than 98%). In the group with nearly normal semen analysis values the frequencies of spermatozoa disomic for chromosome 1 or chromosome 7 were 0.08% and 0.07%, respectively, and not elevated compared to controls (0.10% and 0.06%, respectively). The frequency of diploid spermatozoa was 0.17%, not significantly different from the control group (0.15%) either. In the group of oligoastheno teratozoospermic men both the frequencies of disomic cells for chromosome 1 (0.22%) and for chromosome 7 (0.13%) and of diploid spermatozoa (0.56%) were significantly higher compared to controls, although this was mainly due to one patient with high frequencies of hyperploid sperm. The results indicate that infertility may be a risk factor for chromosomal aneuploidy in spermatozoa. PMID- 9215781 TI - An unusual radiological finding in thanatophoric dysplasia type 1 with common mutation of the fibroblast growth factor receptor-3 (FGFR3) gene (Arg248Cys) PMID- 9215782 TI - Hyperfractionated and accelerated radiation therapy in central nervous system tumors (malignant gliomas, pediatric tumors, and brain metastases). AB - The authors review the main contributions of the international literature concerning the role of hyperfractionation (HF), accelerated fractionation (AF), and accelerated hyperfractionation (AHF) of the dose in radiation therapy (RT) of central nervous system tumors. Basic rationales, clinical results, acute/late toxicity, and current prospectives are summarized in three sections focusing on malignant gliomas, pediatric brainstem tumors, and brain metastases. In supratentorial malignant gliomas the superiority of AHF (0.89 Gy x 3 fractions/day; total dose 61.4 Gy) over conventional fractionation ((CF) total dose 58 Gy) was demonstrated by a randomized trial. However, the gain in median survival time was less than 6 months. No other randomized trials support the preferential choice of non-CF schedules outside clinical trials. Ongoing trials are exploring the role of AHF in combination with chemotherapy, hypoxic cell and radiosensitizing agents. As for pediatric brainstem tumors, there are no data to support the routine use of HF that should be preferably used in an investigative setting. As late sequelae have been reported in the few long-term survivors, patients should be carefully selected. Regarding brain metastases AF RT and AHF RT, with their faster treatment course, may represent a convenient alternative to CF RT for the palliation of brain metastases. In carefully selected patients with solitary brain metastases non-CF RT may be part of aggressive treatment approaches. PMID- 9215783 TI - Primary lymphoma of brain: results of management of a modern cohort with radiation therapy. AB - PURPOSE: To assess the outcome and prognostic factors for patients with primary lymphoma of brain managed with radiation therapy between 1979 and 1988. METHODS AND MATERIALS: A retrospective review was undertaken of 49 patients referred to Princess Margaret Hospital. There were 25 males and 24 females. Median age was 60 years, with a range of 17-80 years. Tumors were located supratentorially in 35, infratentorially in 10, and both in 4 patients. Single masses were demonstrated on CT brain in 36, and multiple lesions in 13 patients. Cranial irradiation was given in 48, and 11 patients received chemotherapy. All patients in this series were immunocompetent. RESULTS: Over a follow-up range of 3-11 years of surviving patients, with a median of 6 years, 40/49 patients have died. Overall median survival was 1.4 years (17 months) and 5-year actuarial survival was 26%. Statistical analysis revealed the following significant factors: Karnofsky performance status (KPS), age, and distribution pattern of disease on presenting CT brain. Five-year actuarial survival for patients with a KPS > 60 or <60 was 56% and 10%, respectively (P = 0.01); for patients with age < 60 or >60, 5-year actuarial survival was 42% and 9%, respectively (P = 0.03); for patients with solitary or multiple lesions, 5-year actuarial survival was 30% and 15%, respectively (P = 0.04). CONCLUSIONS: We conclude that Karnofsky performance status, age, and distribution pattern on pretreatment CT of brain are significant prognostic factors in primary lymphoma of brain, and that new approaches need to be developed for these patients. PMID- 9215784 TI - The risk of interstitial radiotherapy of low-grade gliomas. AB - BACKGROUND AND PURPOSE: The risk of side effects of low activity (i.e. <20 mCi) Iodine-125I (125I) interstitial radiotherapy was analyzed in patients with low grade gliomas. MATERIALS AND METHODS: Permanent (247 patients) or temporary 125I implants (268 patients) were used with a median reference dose of 60 Gy and 100 Gy, respectively, which was calculated to the outer rim of the tumour. The mean dose rate for temporary implants was low (median, 10 cGy/h). Risk factors were obtained from the multivariate proportional-hazards model. RESULTS: Radiogenic complications occurred in 39/515 patients (28 patients with transient symptoms and 11 patients with progressive symptoms). The most important risk factor was the volume of the intratumoural 200 Gy isodose. Available experimental data have associated a high dose zone in this range with the size of the treatment induced radionecrosis. Rapid tumour shrinkage (decrease of the tumour volume > or =50%) within the first 6 months with subsequent centripetal movement of non-pathologic tissue into the high dose zone and a reimplantation were additional risk factors. Radiation injury after rapid tumour shrinkage could be better avoided with temporary implants. A 200 Gy isodose volume <4.5 ml corresponded to an estimated risk of radiogenic complications <3%. There was a steep increase of the risk beyond this limit. Translation of the 200 Gy isodose volume in terms of the treatment volume and the reference dose allows rational treatment planning. The estimated risk of a temporary implant with an applied reference dose of 60 Gy and a treatment volume <23 ml was <3%. CONCLUSIONS: The intratumoural necrotizing effect of a low activity 125I implant limits its application to small treatment volumes. Radiation injury outside the treatment volume can be better avoided with temporary implants in the case of rapid tumour shrinkage. PMID- 9215785 TI - Contrast-enhanced magnetization transfer imaging: improvement of brain tumor conspicuity and delineation for radiosurgical target volume definition. AB - PURPOSE: To assess the contrast-noise-ratio (CNR), and thus tumor conspicuity and delineation, on contrast-enhanced T1-weighted magnetization transfer (MT) images compared to conventional T1-weighted spin echo (SE) images as a strategy to improve definition of the macroscopic boost volume in radiosurgery treatment planning in patients with high grade gliomas or metastatic brain lesions. MATERIALS AND METHODS: Fifty patients (mean age, 51 years) with histologically proven or suspected high grade glioma or cerebral metastases were prospectively examined by MR imaging. Following gadolinium dimeglumine administration (0.1 mmol/kg body weight) the brain was imaged with both a T1-weighted MT-fast low angle shot (FLASH) pulse sequence and with a conventional T1-weighted SE sequence without MT saturation. Lesion conspicuity, size and CNR were compared for both techniques. RESULTS: The mean tumor diameter of malignant gliomas was significantly (P < 0.01) larger when measured on T1-weighted MT-FLASH images compared to T1-weighted SE images and was comparable for metastatic lesions. The mean CNR of enhancing lesions on T1-weighted MT-FLASH was 14 +/- 5 compared to 10 +/- 4 on SE images, representing a significant (P < 0.05) improvement. Lesion conspicuity and delineation was improved in 10 of 20 patients (50%) with high grade gliomas and in 15 of 30 patients (50%) with metastases. Additional contrast enhancing lesions were detected in 8 of 30 patients (27%) with metastases on MT FLASH images. Lesion conspicuity was markedly improved in the posterior fossa. DISCUSSION: Contrast-enhanced T1-weighted MT-FLASH images improve lesion detection and delineation in the planning process of radiosurgery in patients with intracranial high grade gliomas or metastases and may even alter the treatment approach. PMID- 9215786 TI - Efficacy and feasibility of stereotactic radiosurgery in the primary management of unfavorable pediatric ependymoma. AB - Stereotactic radiosurgery was given as a boost in the initial radiation management of five children with localized intracranial ependymoma. Preliminary results in young children with high-risk tumors indicate good local control without excessive neurotoxicity. PMID- 9215788 TI - A simple method for 3D lesion reconstruction from two projected angiographic images: implementation to a stereotactic radiotherapy treatment planning system. AB - INTRODUCTION: The most used imaging modality for diagnosis and localisation of arteriovenous malformations (AVMs) treated with stereotactic radiotherapy is angiography. The fact that the angiographic images are projected images imposes the need of the 3D reconstruction of the lesion. This, together with the 3D head anatomy from CT images could provide all the necessary information for stereotactic treatment planning. We have developed a method to combine the complementary information provided by angiography and 2D computerized tomography, matching the reconstructed AVM structure with the reconstructed head of the patient. MATERIALS AND METHODS: The ISIS treatment planning system, developed at Institute Curie, has been used for image acquisition, stereotactic localisation and 3D visualisation. A series of CT slices are introduced in the system as well as two orthogonal angiographic projected images of the lesion. A simple computer program has been developed for the 3D reconstruction of the lesion and for the superposition of the target contour on the CT slices of the head. RESULTS AND CONCLUSIONS: In our approach we consider that the reconstruction can be made if the AVM is approximated with a number of adjacent ellipses. We assessed the method comparing the values of the reconstructed and the actual volumes of the target using linear regression analysis. For treatment planning purposes we overlapped the reconstructed AVM on the CT slices of the head. The above feature is to our knowledge a feature that the majority of the commercial stereotactic radiotherapy treatment planning system could not provide. The implementation of the method into ISIS TPS shows that we can reliably approximate and visualize the target volume. PMID- 9215787 TI - Factors influencing the risk for complications following Gamma Knife radiosurgery of cerebral arteriovenous malformations. AB - BACKGROUND AND PURPOSE: We reported previously a model predicting the risk for radiation-induced complications following Gamma Knife radiosurgery for AVM. No factor other than the dose distribution was related to the risk. The aim of this study was to define if other parameters are of importance for the risk of complications. MATERIAL AND METHODS: The model above was used to calculate the risk for complications in all 1128 AVM patients Gamma Knife-treated at the Karolinska Hospital 1970-1993. The number of predicted complications was compared to the number of observed ones for a number of different parameters. RESULTS: The model underestimated the risk of complications for patients previously given radiation with multiple or single fractions. Neither age nor gender influenced the risk of complications. Centrally located AVM had a higher, and peripheral a lower incidence of complications as compared to the calculated risk, and a previous hemorrhage reduced the risk of complications. From the observed number of complications, parameters in the model were determined by a fitting procedure separately for three groups of AVM: central and peripheral with and without a previous hemorrhage. It is also shown that the assumption of a serial functional architecture is valid in the model. This was investigated by the use of a relative seriality model with a combined serial-parallel functional architecture. CONCLUSIONS: The risk of complications following radiosurgical treatment of AVM is dependent on the clinical history, AVM location and whether the patient has received radiation earlier. PMID- 9215789 TI - Dosimetric and cytogenetic studies of multiple radiation-induced meningiomas for a single patient. AB - No criteria are currently available to determine the spontaneous or radiation induced origin of a malignant tumor occurring in a previously irradiated area. This study presents the dosimetric and cytogenetic analysis of meningiomas diagnosed in irradiated brain areas from a single patient and a discussion of the karyotypes of spontaneous meningiomas and radiation-induced tumors published in the literature. PMID- 9215790 TI - Response of human hair cortical cells to fractionated radiotherapy. AB - Hair cortical cell counting (HCCC) represents a non-invasive, in-vivo measure of cell kill in the human integument. Sixty-six patients undergoing conventionally fractionated, external beam radiotherapy for early stage carcinoma of the prostate had groin hair samples counted. This technique is a sensitive and reproducible measure of radiation effect and may have applicability as an in-vivo prediction tool or in the field of biological dosimetry. A repopulative follicular response occurring at 3-4 weeks may explain flattening of the dose response curve. PMID- 9215791 TI - Synergistic cytotoxic effects of ether phospholipid analogues and ionizing radiation in human carcinoma cells. AB - BACKGROUND AND PURPOSE: There is growing evidence in recent years that the antiproliferative effects of ionizing radiation may not be exclusively mediated via DNA damage but also by interactions and alterations of cell membrane associated processes. Here, we tested the hypothesis that membrane active cytotoxic ether lipids and analogues may interact with ionizing radiation, enhancing its antiproliferative effects. MATERIALS AND METHODS: The two epithelial tumor cell lines HTB 43 and KB, and the ether lipid resistant subline KBr were treated by a combination of radiation and ether lipids. Cytotoxic effects were measured by colony forming assays and the effects on membrane phospholipids were determined by quantitative thin-layer chromatography of cell lipid extracts. RESULTS: We present evidence that some ether lipids show supra additive cytotoxic effects with ionizing radiation. These effects seem to depend on the same structural properties of ether lipids that determine their intrinsic cytostatic and cytotoxic activity. Identical growth inhibitory results were achieved when cells were treated before, or 30 min after irradiation. Analysis of major membrane phospholipids revealed no statistically significant differences of phospholipid distribution pattern in cells exposed to both treatment modalities. CONCLUSION: Our data indicate that changes of overall membrane phospholipid composition do not seem to be the mechanism of synergistic antiproliferative activity of ether lipids and ionizing radiation. PMID- 9215792 TI - Exploration of new treatment modalities offered by high energy (up to 50 MeV) electrons and photons. AB - BACKGROUND AND PURPOSE: A number of deep seated tumours are difficult to treat conformally with photon beams mainly due to the almost exponential dose decrease with depth. MATERIALS AND METHODS: In order to improve the conformity of these treatments a number of useful characteristics of high energy (above 20 MeV) electron beams of the MM50 Racetrack Microtron have been systematically investigated and clinically applied. RESULTS: A typical characteristic of electron beams with energies up to 20 MeV is the sharp dose fall-off with depth. At higher energies this effect is less pronounced but may be improved by adding a small fraction of photons with a matching dose gradient (wedge). With this technique, high energy electrons can be used close to sensitive organs down to 17 cm depth. Another physical characteristic of high energy electrons is the sharp penumbra at depths down to 4-5 cm and the possibility to use opposed electron beams in order to enhance the dose centrally or near the centre of a body. Skin sparing by delivering a part of the absorbed dose with photons through the same beam portal as the electrons has also been systematically studied. These characteristics of the high-energy electron beams have been utilised in the optimisation of some clinical treatments. CONCLUSIONS: Electron beams in this high energy region give increased possibilities to achieve dose conformity. Enhanced conformity can be obtained especially if electrons and photons are combined to augment some specific characteristics of the electron beams. PMID- 9215793 TI - Depth for dose calibration in high energy photon beams. AB - BACKGROUND AND PURPOSE: The normalisation depth for determination of output factors in photon fields has frequently been the depth of dose maximum. At high energies the contribution from contaminating electrons is significant at dose maximum and is critically dependent on the beam geometry parameters, which is why a larger depth should be preferred. MATERIALS AND METHODS: The effect of electron contamination was studied using a purging magnet to remove charged particles from the treatment head and a helium bag to minimise production between the head and the phantom. RESULTS: A depth of 10 cm was found to be beyond the range of the contaminating electrons for photon energies up to 20 MV (TPR(20)(10) = 0.772). However, at 50 MV (TPR(20)(10) = 0.810) contaminating electrons contribute 2-3% to the absorbed dose at 10 cm depth. CONCLUSIONS: 10 cm is recommended as both reference and normalisation depth for all megavoltage photon beam qualities, i.e. 60Co and X-rays from accelerators up to 50 MV. PMID- 9215794 TI - Three dimensional variability in patient positioning using bite block immobilization in 3D-conformal radiation treatment for ENT-tumors. AB - BACKGROUND AND PURPOSE: The aim of this prospective study was to analyze the three-dimensional (3D) reproducibility of the isocenter position and of patient positioning with the use of bite block immobilization by means of a simple verification procedure for a complex beam arrangement applied for ENT-tumors. MATERIALS AND METHODS: We analyzed the positioning data of 29 consecutive patients treated for ENT-tumors at the Department of Radiotherapy and Oncology of the University of Wurzburg. A total of 136 treatment sessions were analyzed. Patients were positioned and immobilized using an individualized bite block system and a head and neck support. A complex beam arrangement was applied combining two offset rotational and two oblique wedge fields on a 5 MV linear accelerator. Orthogonal verification films were taken once weekly. Four to six film pairs per patient were obtained (during 4-6 weeks) with a mean number of 4.7 film pairs per patient. These were compared to the corresponding orthogonal simulator films taken during primary simulation. Deviations of the verified isocenter from the isocenter on the simulator film were measured and analyzed in three dimensions in terms of overall, systematic and random categories. A 3D deviation vector was calculated from these 3D data as well as a 2D-deviation vector (for comparison with literature data) from the lateral verification films. RESULTS: The overall setup deviation showed standard deviations (SD) of 2.5, 2.7 and 3.1 mm along the cranio-caudal, anterior-posterior and medio-lateral axes, respectively. The random component ranged from SD 1.9 to 2.1 mm and the systematic component ranged from SD 1.8 to 2.2 mm. The mean length of the 3D vector was 3.1 mm for the systematic as well as the random component. Ninety percent of 3D systematic and random deviations were less than 5 mm. The mean length of the 2D-vector was 2.4 mm for the random component and 2.2 mm for the systematic component. Ninety percent of 2D-random and systematic variations were less than 4 mm. CONCLUSIONS: The presented individualized bite block immobilization device provides an accurate and reproducible patient positioning for 3D-conformal radiation therapy in the head and neck. Random and systematic deviations in each of the three directions are in the range of +/-4 mm (2 SD, comprising 95% of the deviations) and are within the range or even less than deviations described for most thermoplastic or PVC-mask fixation devices. These deviations should be taken into account during definition of planning target volume in head and neck tumors. PMID- 9215795 TI - European historical note of intraoperative radiation therapy (IORT): a case report from 1905. PMID- 9215796 TI - Accelerated radiotherapy with carbogen and nicotinamide (ARCON) in high grade malignant gliomas. PMID- 9215797 TI - Is there sufficient evidence of hypersensitivity to low doses in radiotherapy? PMID- 9215798 TI - In vivo electron paramagnetic resonance studies on oxidative stress caused by X irradiation in whole mice. AB - The effect of x-irradiation on the reduction rates of nitroxyl radicals was examined in whole mice using in vivo EPR. One hour after irradiation, the reduction rates of nitroxyl increased up to 15 Gy irradiation, but decreased over this dose. The enhancement of the reduction rate of nitroxyl was suppressed by preadministration of a radioprotector, cysteamine, suggesting that the enhancement of nitroxyl reduction is related to the radiation damage. Thiobarbituric acid-reactive substances (TBARS) in liver homogenate were increased by x-irradiation, indicating that x-irradiation induced oxidative stress in mice. Endogenous antioxidant, alpha-tocopherol, and the activities of antioxidative enzymes such as superoxide dismutase (SOD), catalase, and glutathione peroxidase were not induced by x-irradiation under these experimental conditions. Eventually the nitroxyl reduction in whole mice should be enhanced by the oxidative stress due to x-irradiation. An in vivo EPR system probing the nitroxyl reduction should be applicable to the noninvasive study on the oxidative stress caused by radiation. PMID- 9215799 TI - Impaired cellular cholesterol efflux by oxysterol-enriched high density lipoproteins. AB - One of the proposed antiatherogenicity role of high-density lipoproteins (HDL) is believed to stimulate removal of cholesterol from the peripheral cells back to the liver for excretion. We have investigated the effects of oxidation-related modifications of HDL on their ability to stimulate cholesterol efflux from cultured cells. Human HDL (HDL3, 1.13 < d < 1.21 g/ml) have been modified either by malondialdehyde or by copper-mediated oxidation (Ox-HDL3). Compared with native HDL3, the modified HDL3 resulted in a significantly reduced efflux of labeled cholesterol from preloaded macrophages (P388D1 cell line). Analysis of lipid composition of Ox-HDL3 by gas chromatography revealed the presence of oxysterols (OS). Enrichment of native HDL3 with oxysterols resulted in a reduced capacity to stimulate cholesterol efflux. The reduced ability of OS-enriched HDL3 to elicit cholesterol efflux may contribute to cellular cholesterol accumulation and subsequently to atherosclerosis. PMID- 9215800 TI - Oxidation of aminopyrine by the hydroperoxidase activity of lipoxygenase: a new proposed mechanism of N-demethylation. AB - The oxidation of aminopyrine, an N-alkyl aromatic amine, by the hydroperoxidase activity of lipoxygenase was studied. Aminopyrine gave rise to a purple color in the presence of H2O2 and lipoxygenase, the color being proportional to the aminopyrine radical cation. The H2O2/aminopyrine radical cation molar ratio was 0.5. The overall reaction was considered as an enzymic-chemical second order mechanism with substrate regeneration. From the equations, the apparent constant of the radical cation's decomposition (k'app) was evaluated under different experimental conditions. It was found to be inversely proportional to the proton concentration but unaffected by the concentration of aminopyrine. These results suggest a new comprehensive mechanism for N-demethylation, which takes into account the described presence of both nitrogen- and carbon-centered radicals and the marked effect of pH on the stability of the radical cation. PMID- 9215801 TI - Oxidation of nuclear membrane cholesterol inhibits nucleoside triphosphatase activity. AB - Oxygen derived free radicals can oxidize membrane cholesterol. We have previously shown that cholesterol in the nuclear membrane can modulate nuclear nucleoside triphosphatase (NTPase) activity. Nucleocytoplasmic transport of peptides and mRNA via the nuclear pore complex may be regulated by the NTPase. The purpose of the present study was to determine if oxidation of nuclear cholesterol could alter NTPase activity. Nuclear membrane cholesterol was oxidized in situ with cholesterol oxidase (to selectively oxidize cholesterol) and NTPase activity measured. HPLC analysis confirmed the formation of cholesterol oxides. The activity of the NTPase was strikingly inhibited by cholesterol oxidase treatment. The Vmax of the NTPase was significantly decreased after cholesterol oxidase treatment but the Km value was unchanged. The sensitivity of NTPase activity to varying cholesterol oxidase concentrations also suggested that cholesterol located in the inner leaflet of the nuclear membrane appeared to be more important in the modulation of NTPase activity than that in the cytoplasmic leaflet. Our results indicate that oxidation of nuclear membrane cholesterol inhibits NTPase activity. These results have implications for peptide and mRNA flux across the nuclear membrane during conditions where lipid oxidation may be expected. PMID- 9215803 TI - Clastogenic factors in plasma of HIV-1 infected patients activate HIV-1 replication in vitro: inhibition by superoxide dismutase. AB - The frequent neoplastic disorders present in HIV-infected patients and the implication of oxidative stress in AIDS-Kaposi's sarcoma pathogenesis prompted us to study whether the mechanisms implicated in genotoxic effects of clastogenic factors (CFs) (i.e., chromosome damaging materials released by cells under conditions of oxidant stress) can play a role in HIV-1 expression and whether exogenous superoxide dismutase can inhibit the clastogenic and HIV-inducing effects of CFs. CFs were found in the plasma of all HIV-1 infected patients (n = 21) of this study group, in asymptomatic (CDC II) as well as in symptomatic patients (CDC IV). In addition to their chromosome damaging effect, CFs are able to upregulate HIV-1 expression in U1 cells and in PBMCs activated with PHA and IL2 at all time points (p < .05). Their formation, therefore, is an early event in the disease. It occured despite antiviral medication in these patients. Superoxide dismutase inhibited the clastogenic and the viral inducing effects (p < .05). On the basis of our findings, association of SOD mimetics or superoxide scavengers with antiviral drugs may be a new therapeutic approach. This polytherapy, if started early enough after infection, may prolong the latency period and limit the emergence of drug-resistant viral strains. PMID- 9215802 TI - Glutathione dependent reduction of alloxan to dialuric acid catalyzed by thioltransferase (glutaredoxin): a possible role for thioltransferase in alloxan toxicity. AB - Recombinant pig liver thioltransferase (rPLTT) catalyzes the reduction of alloxan to dialuric acid by glutathione (GSH). This is the second non-disulfide substrate, after dehydroascorbic acid, described for thioltransferase. The reaction kinetics, measured by a coupled assay including glutathione disulfide reductase and NADPH yielded a Km = 82 microM for alloxan, a k(cat) = 37 s(-1), and a k(cat)/Km = 4.5 x 10(5) M(-1) s(-1). The presence of rPLTT suppressed the competitive formation of compound 305, an alloxan-GSH conjugate of unknown structure, and at GSH concentrations between 0.05 mM and 1.5 mM, oxygen consumption was greater than that recorded in the uncatalyzed reaction. Both superoxide dismutase and catalase inhibited oxygen consumption in 1.0 mM GSH and 0.2 mM alloxan in the presence of rPLTT. This study suggests that thioltransferase (glutaredoxin) plays a significant role in the cytotoxicity of alloxan in vulnerable tissues. PMID- 9215805 TI - Increased ascorbate radical formation and ascorbate depletion in plasma from women with preeclampsia: implications for oxidative stress. AB - There is evidence that oxidative stress accompanies preeclampsia and plasma ascorbate concentrations are reported to be decreased in the disorder. We tested the hypothesis that an ascorbate-oxidizing activity is increased in plasma from women with preeclampsia relative to normal pregnancy. Electron paramagnetic resonance (EPR) spectroscopy was used to determine (1) plasma functional reserves of ascorbate and total thiols, (2) temporal changes in ascorbate and thiol concentrations during incubation of whole blood in vitro, and (3) ascorbate radical signal kinetics in plasma after equalization of ascorbate concentrations. High-pressure liquid chromatography (HPLC) was used to measure plasma alpha tocopherol. Ascorbate concentrations were 50% lower in preeclampsia relative to normal pregnancy plasma but thiols and alpha-tocopherol did not differ. The elapsed time prior to half-consumption of plasma ascorbate was decreased approximately three-fold during incubation of whole blood from preeclamptics. No concomitant decrease in thiols was evident. The initial ascorbate radical signal amplitude was greater in preeclampsia plasma and then, in contrast to normal pregnancy plasma, decreased progressively. The iron chelator, deferoxamine had no effect on plasma ascorbate radical formation. We conclude that an ascorbate oxidizing activity is increased in preeclampsia plasma which might contribute to vascular dysfunction in the disorder. PMID- 9215804 TI - Peroxidase-dependent bioactivation and oxidation of DNA and protein in benzo[a]pyrene-initiated micronucleus formation. AB - Micronucleus formation initiated by benzo[a]pyrene (B[a]P) and related xenobiotics is widely believed to reflect potential carcinogenic initiation, yet neither a dependence upon bioactivation nor the critical enzymes have been demonstrated. Using rat skin fibroblasts, protein oxidation (carbonyl formation) and content of prostaglandin H synthase (PHS) and cytochrome P4501A1 (CYP1A1) protein were determined by Western blot/immunodetection with enhanced chemiluminescence. DNA oxidation as 8-hydroxy-2'-deoxyguanosine formation was quantified using high-performance liquid chromatography with electrochemical detection. Fibroblast CYP1A1 activity assessed as ethoxyresorufin-O-deethylase was not detectable, and even CYP1A1 protein was measurable only after induction with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). However, TCDD additionally induced prostaglandin H synthase (PHS), which also was detectable constitutively. B[a]P 10 microM initiated the oxidation of DNA and protein, and the formation of micronuclei, all of which were enhanced over 2-fold by the dual CYP1A1/PHS inducer TCDD 10 nM, as well as by other PHS inducers, 12-O-tetradecanoylphorbol 13-acetate 1 microM and interleukin-1alpha 0.625 or 1.25 ng/ml, that do not induce CYP1A1 (p < .05). Conversely, B[a]P target oxidation and micronucleus formation were abolished by 1-aminobenzotriazole 1 mM (p < .05), which was a potent inhibitor of both peroxidases and P450. These results provide the first direct evidence that B[a]P-initiated micronucleus formation, like carcinogenic initiation, requires enzymatic bioactivation, and that peroxidase-dependent, reactive oxygen species-mediated oxidation of DNA, and possibly protein, constitutes a molecular mechanism of initiation in uninduced cells. Induction of either CYP1A1 or peroxidases such as PHS substantially enhances this genotoxic initiation, which may reflect cancer risk. PMID- 9215806 TI - 4-Hydroxy-2-nonenal hardly affects glycolysis. AB - 4-Hydroxy-2-nonenal (HNE), one of the major products of lipid peroxidation, inactivated the rate-limiting enzymes (from animal sources) of the glycolytic pathway and the pentose phosphate pathway when incubated at 37 degrees C for 1 h in the absence of glutathione (GSH). The HNE concentration for half-maximal inactivation of 6-phosphofructokinase (PFK) and glyceraldehyde-3-phosphate dehydrogenase was 3-10 microM; and that value for pyruvate kinase, glucose-6 phosphate dehydrogenase, and hexokinases I and II was 0.15-0.6 mM. In the presence of 5 mM GSH, however, only PFK, irrespective of the source (muscle, liver, or erythrocyte), was inactivated by 40-50% when incubated with 0.1 mM HNE for 1 h. Even PFK was not inactivated in the presence of both GSH and its substrate, ATP (2 mM). Glycolysis in human erythrocytes was not affected by treatment of cells with 0.1 mM HNE at 37 degrees C for 30 min. The results suggest that HNE, at concentrations observable under physiological and pathological conditions, hardly affects glycolysis in cells. PMID- 9215807 TI - Photo-oxidative disruption of lysosomal membranes causes apoptosis of cultured human fibroblasts. AB - Acridine orange (AO) is a lysosomotropic weak base, a metachromatic fluorochrome, and a photosensitizer, as well. Living cells that are exposed for a short period of time to this compound at low concentration, and under ordinary culture conditions, accumulate the drug within their acidic vacuolar compartment, giving rise to a mainly red, granular fluoresence upon excitation with blue light. When AO-loaded cells are irradiated with intense blue light, AO soon starts to leak from late endosomes and lysosomes, partially shifting the fluorescence to a green, nuclear and diffuse cytosolic, one. This AO-relocalization is a consequence of photo-oxidation of the lysosomal membranes, which initially results in disruption of their proton-gradients and later, in leakage into the cytosol of a host of hydrolytic enzymes--as was here demonstrated by immunocytochemistry--which are capable of causing cellular damage. Most fibroblasts survived minor photo-oxidation, with a period of reparative autophagocytosis. Severe photo-oxidation, which resulted in severe lysosomal damage, caused cellular necrosis; whereas moderate stress, resulting in only partial lysosomal leakiness lead to apoptosis with TUNEL-positive nuclei and shrunken cytoplasm. The findings of the present study show that photo-oxidative damage to the membranes that surround the acidic vacuolar compartment, is an event that results in release of proteolytic and DNA-fragmenting enzymes into the cytosol, which may induce either necrosis, apoptosis, or reparable sublethal damage, depending on the magnitude of lysosomal rupture. Furthermore, the results strongly suggest that proteases and endonucleases of lysosomal origin may induce apoptosis if relocalized from the acidic vacuolar compartment into the cytosol. PMID- 9215808 TI - Role of rpoS regulon in resistance to oxidative stress and near-UV radiation in delta oxyR suppressor mutants of Escherichia coli. AB - Escherichia coli delta oxyR mutants are hyper-sensitive to oxidative agents but this sensitivity is reversed to hyper-resistance in delta oxyR suppressor strains (delta oxyRsup; Greenberg, J.T. and Demple, B. 1988. EMBO J. 7:2611-2618). Also, delta oxyR mutants have increased mutation rates that are also reversed in delta oxyRsup. We now report that the rpoS regulon may have a role in determining hyper resistance and loss of hyper-mutability of delta oxyRsup. Delta oxyRsup cells were also resistant to near-ultraviolet radiation (near-UV) and survived longer in stationary phase than delta oxyR cells. In delta oxyRsup cells elevated beta galactosidase expression from a rpoS::lacZ promoter fusion and significant overproduction of RpoS protein was observed. These increases were accompanied by substantial elevation in transcription of rpoS-dependent genes as determined by beta-galactosidase expression from katE::lacZ, dps::lacZ, and xthA::lacZ promoters. Catalase HPI and HPII activities were also increased. When rpoS::Tn10 was transduced into delta oxyRsup, phenotypes switched back to hyper-sensitive, hyper-mutable and reduced catalases I and II. Individual delta oxyR colonies exhibited significant clonal variability in beta-galactosidase expression from rpoS::lacZ promoter. These results provide further evidence of the functional and regulatory overlap between two major anti-oxidant defense systems of bacteria. PMID- 9215809 TI - Glutamate toxicity on a PC12 cell line involves glutathione (GSH) depletion and oxidative stress. AB - The effect of antioxidants and reducing agents on glutamate-induced cytotoxicity was examined using PC12 cells. The antioxidants vitamin E, idebenone, and selegiline protected cells against the cytotoxicity observed 24 h after exposure to 0.5 or 10 mM glutamate, as determined by lactate dehydrogenase leakage, even when added 3 h after glutamate. The reducing agents, glutathione (GSH) and dithiothreitol (DTT), also provided protection against the cytotoxicity of glutamate. Preincubation of PC12 cells with the antioxidants mentioned above, or the incubation with those antioxidants after exposure to glutamate for 3 h, prevented the reduction of viability caused by glutamate. Cystine uptake was inhibited by exposure of cells to glutamate, as determined by L-[35S]-cystine uptake. Incubation of cells with 0.5 or 10 mM glutamate caused a marked decrease in cellular GSH levels, not prevented by antioxidants. The activity of GSSG reductase was decreased by glutamate and this inhibition was reverted in the presence of the reducing agents GSH and DTT. These results indicate that glutamate toxicity on PC12 cells results from the inhibition of cystine uptake with consequent GSH depletion and oxidative stress, suggesting that antioxidants may reduce the cellular damage in pathologic conditions associated with excessive glutamate release. PMID- 9215810 TI - Gender and maturation affect glutathione status in human neonatal tissues. AB - Gender and maturation affect glutathione status in human neonatal tissues. The objective was to verify if human tissues derived from baby girls had a greater ability then tissues derived from males to stimulate the glutathione-reductase, when faced with an oxidative challenge. In vitro, the effect of a calibrated oxidative challenge was studied in endothelial cells. In vivo, the effect of a clinically relevant oxidative challenge was studied in cells from tracheal aspirates derived from oxygen-dependent newborn infants. In endothelial cells, the oxidant tert-butylhydroperoxide had a stimulating effect on GSSG-R activity in cells derived from females. The peroxide produced a time, concentration and gender-dependent cytotoxicity, with female-derived cells exhibiting a better viability. In vivo, the intracellular total glutathione content was higher in female-derived cells and in cells from more mature babies; postnatal age and gestational age had a positive effect on the activity of GSSG-R. Oxygen (FiO2 > or = 0.3) was associated with a lower activity of GSSG-R in boys, early in life. Considering that glutathione is a central element in the antioxidant defense, these results suggest that specific tissues derived from the baby girl are potentially better protected against an oxidative stress than those derived from the boy. PMID- 9215811 TI - Oxidized lipoproteins including HDL and their lipid peroxidation products inhibit TNF-alpha secretion by THP-1 human macrophages. AB - It has been established that oxidized LDL (ox-LDL) modifies cytokine secretion by macrophages, for example, by reducing tumor necrosis factor alpha (TNF-(alpha) m RNA. However, little is known about the effects of oxidized high density lipoprotein (ox-HDL). This study reports the effects of ox-HDL subfractions 2 and 3 (ox-HDL2, ox-HDL3) compared with that of ox-LDL and some products of oxidation (hydroperoxides and aldehydes) on the secretion of TNF-alpha from THP-1 human monocytes derived macrophages in vitro. HDL2, HDL3 and LDL were oxidized with 10 microM Cu++ for 12 h and/or 24 h. Native and oxidized HDL and LDL were incubated for 24 h with macrophages with or without LPS (10 ng/ml) after which TNF-alpha secretion was measured in the culture medium. Lipid hydroperoxides and apolar aldehydes were also incubated with the cells for 2 h following which the medium was replaced and TNF-alpha secretion measured after a further 22 h of incubation. An inhibition of TNF-alpha by ox-HDL2 (p < .05), ox-HDL3 (p < .05) and ox-LDL (p < .05) from THP-1 macrophages was observed in the presence and absence of LPS. This inhibition remained the same after incubation with ox-HDL 12 h and 24 h. Hydroperoxides of linoleic acid did not modify TNF-alpha secretion by cells while five out of eight aldehydes analyzed (2,4-heptadienal, hexanal, 2-nonenal, 2 octenal, 2,4-decadienal) inhibited TNF-alpha secretion (p < .05). These findings demonstrate that ox-HDL, and some of its lipid peroxidation products, plays a role in the modulation of the inflammatory response by macrophages as previously observed for ox-LDL. PMID- 9215812 TI - How does superoxide dismutase protect against tumor necrosis factor: a hypothesis informed by effect of superoxide on "free" iron. AB - The manganese-containing mitochondrial superoxide dismutase (MnSOD) is induced by TNF and protects against the necrotic effect of this cytokine. Yet TNF does not increase production of O2- in mitochondria. How is this to be reconciled? TNF is known to increase production of arachidonate, by activation of phospholipase A2 (PLA2). Arachidonate will be converted to the corresponding alkyl hydroperoxide by lipoxygenase. O2- increases "free" iron by oxidizing [4Fe-4S] clusters of dehydratases, such as aconitase. Ferrous iron in turn reacts with alkyl hydroperoxides, in an analogue of the Fenton reaction, to produce alkoxyl radicals: which can initiate the oxidation of polyunsaturated lipids by a free radical chain reaction. MnSOD protects against TNF by decreasing O2- attack on [4Fe-4S] clusters and thus lowering free iron. Inhibitors of PLA2 and of lipoxygenase should also protect by decreasing fatty acyl hydroperoxides and they are known to do so. Cells having little mitochondrial MnSOD, or cells unable to induce that defensive enzyme in response to TNF, will consequently have relatively high levels of "free" iron in that organelle; leading to enhanced lipid peroxidation. Such cells will be preferentially killed by this cytokine. PMID- 9215813 TI - Increase of resting levels of superoxide anion in the whole blood of patients with decompensated liver cirrhosis. AB - The aim of this study is to investigate the relationship between the resting level of superoxide anion (O2.) and liver cirrhosis (LC). The resting levels of superoxide anion in the whole blood of healthy controls and patients with compensated or decompensated LC were measured, by an ultra-sensitive chemiluminescence (CL) analyzer and lucigenin amplification. The assay system can be performed in the absence of leukocyte isolation and stimulant administration. The results showed that the blood CL levels of compensated cirrhotic patients (381.0 +/- 201.5 counts/10 s, mean +/- SD, n = 24) were similar to that of healthy controls (467.9 +/- 299.5 counts/10 s, n = 24). However, the blood CL levels of decompensated cirrhotic patients (2083.5 +/- 1462.4 counts/10 s, n = 24) were significantly greater than that of healthy controls and patients with compensated LC (both p < .001, Student's t-test). The correlation analysis revealed that the blood CL levels in cirrhotic patients were significantly correlated with serum concentrations of albumin (r = -0.65, p < .001) and total bilirubin (r = +0.42, p < .005). However, there was no significant correlation between the blood CL levels and serum levels of transaminases (GOT and GPT). These results suggest that blood levels of superoxide of decompensated cirrhotic patients were greater than those of healthy controls or compensated cirrhotic patients. Moreover, the increase of blood levels of superoxide in decompensated cirrhotic patients is related to the impairment of liver function but not to the inflammation. PMID- 9215814 TI - Lipid peroxidation and modification of lipid composition in an endothelial cell model of ischemia and reperfusion. AB - Among the changes that accompany the development of ischemia are alterations in the composition and turnover of membrane phospholipids. To study these effects, a cell culture model was developed to facilitate accurate measurements of lipids over varying intervals of ischemia and reperfusion (I/R). In order to mimic ischemia, rabbit aortic endothelial cells were grown to confluency on collagen coated beads and the bead cultures allowed to settle to the bottom of a conical test tube or spectrofluorometric cuvette. The cell-coated beads were then resuspended in media to simulate the process of reperfusion. Survival after ischemia/reperfusion, was determined by measurements of cellular replating efficiency, and found to decrease after periods longer than three hours of ischemia (followed by 24 h of reperfusion). Plating efficiencies were reduced to nearly 50% after 5 h of ischemia followed by reperfusion. Release of LDH inversely correlated with cell survival, and lactate production, ATP levels, and extracellular H2O2 concentration were all affected by the duration of ischemia. These changes could be directly related to rates of cellular oxygen consumption which decreased by 50% after 5 h of ischemia, while the percentage of oxygen consumption not be inhibitable by cyanide, increased. Release of esterified fatty acids, which was partly inhibited by the phospholipase A2 inhibitor, mepacrine, was stimulated by increasing periods of ischemia while the incorporation of free fatty acids into phospholipids was inhibited. The incorporation of arachidonic acid was inhibited to a lesser degree than that of oleic or linoleic acids with a resulting change in phospholipid fatty acyl composition favoring greater proportions of unsaturated fatty acids. In some experiments, the effects of vitamin E or ascorbic acid administered prior to ischemia were studied. The degree of fatty acid unsaturation, fatty acid incorporation into phospholipids, and release from phospholipids into the free fatty acid pool during ischemia/reperfusion were not affected by prior administration of vitamin E or ascorbic acid. However, the extent of lipid peroxidation during ischemia was inhibited by 100 mM ascorbic acid when present during the ischemia/reperfusion period, but not by vitamin E administered for 24 h prior to ischemia. Ascorbic acid treatment, but not vitamin E, also enabled cells to recover substantial amounts of the ATP lost following prolonged ischemia. The ATP recovery corresponded to an increased cell survival and decreased lipid peroxidation. Progressive intervals of ischemia followed by reperfusion result in compromised cell respiratory activity and decreased ATP production, and decreased phospholipid acylation leading to net hydrolysis. The associated changes in phospholipid composition, and specifically increased unsaturation appear to favor peroxidation of membrane phospholipids. PMID- 9215815 TI - Preoperative chemotherapy and breast-conserving therapy: why not everyone? PMID- 9215816 TI - Effect of preoperative chemotherapy on local-regional disease in women with operable breast cancer: findings from National Surgical Adjuvant Breast and Bowel Project B-18. AB - PURPOSE: To determine whether preoperative doxorubicin and cyclophosphamide (AC) permits more lumpectomies to be performed and decreases the incidence of positive nodes in women with primary breast cancer. PATIENTS AND METHODS: Women (n = 1,523) were randomized to National Surgical Adjuvant Breast and Bowel Project (NSABP) B-18; 759 eligible patients received postoperative AC and 747, preoperative AC. The clinical size of breast and axillary tumors was determined before each of four cycles of AC and before surgery. Tumor response to preoperative therapy was clinically complete (cCR), partial (cPR), stable (cSD), or progressive disease (cPD). Tissue from patients with a cCR was evaluated for a pathologic complete response (pCR). RESULTS: Breast tumor size was reduced in 80% of patients after preoperative therapy; 36% had a cCR. Tumor size and clinical nodal status were independent predictors of cCR. Twenty-six percent of women with a cCR had a pCR. Clinical nodal response occurred in 89% of node-positive patients: 73% had a cCR and 44% of those had a pCR. There was a 37% increase in the incidence of pathologically negative nodes. Before randomization, lumpectomy was proposed for 86% of women with tumors < or = 2 cm, 70% with tumors 2.1 to 5.0 cm, and 3% with tumors > or = 5.1 cm. Clinical tumor size and nodal status influenced the physician's decision. Overall, 12% more lumpectomies were performed in the preoperative group; in women with tumors > or = 5.1 cm, there was a 175% increase. CONCLUSION: Preoperative therapy reduced the size of most breast tumors and decreased the incidence of positive nodes. The greatest increase in lumpectomy after preoperative therapy occurred in women with tumors > or = 5 cm, since women with tumors less than 5 cm were already lumpectomy candidates. Preoperative therapy should be considered for the initial management of breast tumors judged too large for lumpectomy. PMID- 9215817 TI - Megestrol acetate and aminoglutethimide/hydrocortisone in sequence or in combination as second-line endocrine therapy of estrogen receptor-positive metastatic breast cancer: a Southwest Oncology Group phase III trial. AB - PURPOSE: A phase III randomized trial was performed to determine whether combination hormonal therapy with aminoglutethimide (AG) and hydrocortisone (HC) plus megestrol acetate (MA) improved response rates, response duration, or increased survival over the sequential use of each hormone in women with estrogen receptor-positive metastatic breast cancer (MBC) who had maintained stable disease for at least 6 months or responded to tamoxifen. PATIENTS AND METHODS: Two hundred eighty-eight postmenopausal women with progressive estrogen receptor positive MBC were randomly selected to receive MA 40 mg four times daily (arm I), AG 250 mg four times daily with HC 40 mg daily in divided doses (arm II), versus the combination of MA plus AG given at the same dosages (arm III). Patients on arms I and II who progressed after an adequate trial were crossed over to the other treatment arm. RESULTS: Two hundred thirty-five eligible patients were evaluated for response, time to treatment failure, and survival. Response was only reported for patients with measurable disease and was not statistically different among the three arms. There were two partial responses (PRs) on MA (6%), four complete responses (CRs) and six PRs on AG (24%), and eight PRs and three CRs on MA plus AG (23%) in 32, 42, and 48 measurable patients, respectively. Median times to treatment failure were also similar at 5, 4, and 7 months. Survival was also not statistically different among the three arms at 26, 27, and 26 months for arms I, II, and III, respectively. Toxicity was greater in the two AG arms with respect to fatigue, nausea and vomiting, and rash. CONCLUSION: With the exception of toxicity, there is no response, time to treatment failure, or survival benefit for any one group when comparing MA, AG, or the combination at their stated doses in women with estrogen receptor-positive MBC who had previously responded to or stabilized with tamoxifen. PMID- 9215819 TI - Dose-finding study and pharmacokinetics of epirubicin and paclitaxel over 3 hours: a regimen with high activity and low cardiotoxicity in advanced breast cancer. AB - PURPOSE: To determine the maximum-tolerated dose (MTD) of paclitaxel over 3 hours with a fixed dose of epirubicin, to investigate the plasma pharmacokinetics of this combination, and to evaluate the toxicity and the activity in previously untreated metastatic breast cancer patients. PATIENTS AND METHODS: Fifty patients with metastatic breast cancer, measurable disease, and normal left ventricular ejection fraction (LVEF) were eligible. Epirubicin was administered as an intravenous (I.V.) bolus at the fixed dose of 90 mg/m2 before the infusion of paclitaxel over 3 hours. The initial dose of paclitaxel was 135 mg/m2 and was increased by 20 mg/m2 in subsequent cohorts of six patients until dose-limiting toxicity (DLT). Plasma pharmacokinetics of paclitaxel and epirubicin was performed at cycle 1 in at least two patients per dose level of paclitaxel (175 up to 225 mg/m2). RESULTS: The DLT of this combination was febrile neutropenia in two of eight patients who received paclitaxel at 225 mg/m2. The mean peak plasma concentration of paclitaxel ranged between 5.1 and 6.2 micromol/L at doses of 175 to 225 mg/m2. The concentration of epirubicinol decreased from 47.3 +/- 9.4 to 37.9 +/- 7.5 ng/mL in patients treated with paclitaxel 175 and 225 mg/m2. The most relevant toxicity was grade 4 neutropenia (61% of all courses). The pharmacokinetic data of paclitaxel, in particular the time above the threshold level of 0.05 micromol/L, were not significantly related to myelosuppression. Cardiac toxicity was mild: three patients (6%) developed mild congestive heart failure that was responsive to therapy. Among 49 assessable patients, 41 responses (84%; 95% confidence interval [CI], 70% to 92%) were observed, and nine (18%) of these were complete. CONCLUSION: Our study demonstrates that (1) the MTD is epirubicin 90 mg/m2 and paclitaxel 200 mg/m2; (2) no clear relationship exists between pharmacokinetic data of paclitaxel and myelosuppression, while the increase in the dose of paclitaxel is associated with a reduction in epirubicinol plasma levels; and (3) the association is feasible, with low cardiotoxicity, and has a high activity in metastatic breast cancer. PMID- 9215818 TI - Adjuvant chemotherapy in operable breast cancer: cyclophosphamide, methotrexate, and fluorouracil versus chlorambucil, methotrexate, and fluorouracil--11-year results of Swiss Group for Clinical Cancer Research trial SAKK 27/82. AB - PURPOSE: To compare two adjuvant combination chemotherapies, cyclophosphamide, methotrexate, and fluorouracil (CMF) and chlorambucil, methotrexate, and fluorouracil (LMF), for patients who had undergone potentially curative surgery for unilateral breast cancer, in terms of relapse, survival, and toxicity. PATIENTS AND METHODS: Selection criteria was as follows: stage pT1-3a, N+ or N-, M0, less than 72 years of age. Eligible patients were randomized to receive either CMF (cyclophosphamide 100 mg/m2 orally on days 1 to 14, methotrexate 40 mg/m2 intravenously (I.V.) on days 1 and 8, fluorouracil 600 mg/m2 I.V. on days 1 and 8) or LMF (Leukeran [Wellcome A.G., Bern, Switzerland] 5 mg/m2 orally on days 1 to 14 with the some I.V. cytostatic drugs). Follow-up examinations were performed every 3 months during the first 3 years after mastectomy, and every 6 months thereafter. RESULTS: A total of 246 patients were randomized, of whom 232 who were fully eligible and contribute to the analyses presented here. No statistically significant difference in favor of adjuvant CMF over LMF emerges after a median follow-up duration of 11.2 years, for either overall survival (P = .15) or disease-free survival (P = .14). A consistent trend suggestive of a possible relative benefit associated with CMF should be pointed out. However, CMF presents a significantly worse toxicity profile as concerns hematologic parameters as well as alopecia, nausea, and vomiting. CONCLUSION: This prospective trial has not identified a statistically significant difference in disease-free survival or overall survival between the two adjuvant regimens LMF and CMF. Although a trend in favor of CMF has been observed in premenopausal patients, this has to be weighted against its definitely more pronounced toxicity profile. PMID- 9215820 TI - Prediction of response to antiestrogen therapy in advanced breast cancer patients by pretreatment circulating levels of extracellular domain of the HER-2/c-neu protein. AB - PURPOSE: Overexpression of the HER-2/c-neu/c-erbB2 proto-oncogene is associated with a worse prognosis in patients with breast cancer, perhaps due to an association of the HER-2 proto-oncogene protein with resistance to hormone and/or chemotherapy. Circulating levels of the extracellular domain (ECD) of the HER-2/c neu-related protein (NRP) are elevated in 20% to 40% of patients with metastatic breast cancer. We investigated whether pretreatment levels of NRP predict response to hormone therapy (HT). MATERIALS AND METHODS: Circulating NRP levels were determined in 94 patients who participated in a randomized trial of three different doses of the antiestrogen, droloxifene (DRO), as first-line HT for metastatic breast cancer. RESULTS: NRP levels were elevated (> or = 5,000 U/mL) in 32 of 94 patients (34%). Only three of 32 patients (9%) with elevated NRP levels responded to DRO, compared with 35 of 62 (56%) with nonelevated NRP levels (P = .00001). Low pretreatment NRP level was the most powerful predictor of response to DRO (odds ratio of response, 22.4; P = .0001). Elevated pretreatment NRP levels were also associated with a shorter time to progression (TTP) and survival duration. CONCLUSION: Pretreatment circulating NRP levels predict a low likelihood of benefit from HT, specifically DRO, in patients with estrogen receptor (ER)-positive and/or progesterone receptor (PgR)-positive or receptor unknown metastatic breast cancer, even when adjusted for other known predictive factors, such as ER and/or PgR levels, site of disease, disease-free interval from primary treatment to recurrence, and prior adjuvant chemotherapy. These data suggest that pretreatment NRP levels may be useful in deciding whether to treat a patient who otherwise appears to be likely to respond to HT. PMID- 9215821 TI - Overview of randomized perioperative polychemotherapy trials in women with early stage breast cancer. AB - PURPOSE: To determine whether perioperative polychemotherapy (PeCT) can significantly prolong the overall survival of women with early-stage breast cancer. METHODS: A meta-analysis that used updated individual patient data from all available randomized trials of PeCT, both published and unpublished, was conducted. Data on 6,093 patients (1,124 deaths and 1,912 recurrences) from five clinical trials were available (median follow-up duration, 5.3 years; maximum, 11.3 years). RESULTS: No significant effect of PeCT on overall survival was observed. However, patients who received PeCT had a significantly longer disease free survival (hazards ratio [HR], 0.89; 95% confidence interval [CI], 0.82 to 0.98; P = .02). Time to local recurrence was significantly prolonged in the PeCT arm (HR, 0.68; 95% CI, 0.58 to 0.80; P < .0001). Likewise, there was a borderline significant difference in favor of PeCT in terms of time to distant metastases (HR, 0.90; 95% CI, 0.81 to 1.00; P = .05). Subgroup analyses suggest that node negative women benefited the most from treatment. CONCLUSION: At present, there is no evidence that PeCT is able to prolong overall survival in patients with early-stage breast cancer; however, further follow-up evaluation is required. PeCT significantly prolongs disease-free survival, especially in node-negative women, which emphasizes once more the need for clinical trials in this subgroup. PMID- 9215822 TI - Imaging of estrogen receptors in primary and metastatic breast cancer patients with iodine-123-labeled Z-MIVE. AB - PURPOSE: To evaluate the feasibility of noninvasive imaging of estrogen receptors (ERs) in primary and metastatic breast cancer with the iodine-123-labeled ER specific ligand cis-11beta-methoxy-17alpha-iodovinylestradiol-17beta (Z [123I]MIVE) using conventional nuclear medicine techniques. PATIENTS AND METHODS: Z-[123I]MIVE planar scintigraphy and single-photon emission computed tomography (SPECT) were performed in 12 patients with proven primary breast cancer and 13 patients with proven or from other imaging modalities evident bone, liver, lung, pleura and/or lymph node metastases. The results were compared with those of ER immunohistochemistry (IHC). Blocking studies with the antiestrogen tamoxifen were performed to test whether Z-[123I]MIVE tumor uptake was ER-mediated. RESULTS: Planar imaging showed uptake in 11 of 12 primary carcinomas. ER IHC performed for nine of these was positive. For the planar scintigraphy-negative patient, SPECT was faintly positive, but ER IHC negative (agreement, 90%). In nine of 13 metastatic patients, planar scintigraphy was positive. The agreement between the results of ER IHC on the original primary tumor and of Z-[123I]MIVE scintigraphy was 82%. Specificity of tumor Z-[123I]MIVE uptake was established by complete blockade of uptake by tamoxifen, except in two patients who showed progressive disease. Z-[123I]MIVE scintigraphy also enabled discriminating metastases from confounding nonmalignant abnormalities of the bone scan. CONCLUSION: Z-[123I]MIVE scintigraphy shows high sensitivity and specificity for the detection of ER positive breast cancer. This may have impact on diagnostic possibilities and therapeutic management. Since ER imaging shows the functional status, addressing known intratumoral and intertumoral ER heterogeneity, it may improve the characterization of disease and the selection of patients who may benefit from hormonal therapy. PMID- 9215823 TI - High-dose carboplatin, etoposide, and cyclophosphamide with autologous bone marrow transplantation in first-line therapy for patients with poor-risk germ cell tumors. AB - PURPOSE: A treatment program that included high-dose carboplatin, etoposide, and cyclophosphamide (CEC) followed by autologous bone marrow transplantation (AuBMT) was investigated as first-line therapy in patients with poor-risk germ cell tumors (GCTs). PATIENTS AND METHODS: Previously untreated GCT patients with poor risk features were treated with etoposide, ifosfamide, and cisplatin (VIP) with or without high-dose CEC plus AuBMT. Patients qualified for a change to high-dose CEC if a prolonged clearance of elevated serum tumor markers was observed after two cycles of the cisplatin-containing regimen. RESULTS: Sixteen patients were treated with VIP alone and 14 with VIP and high-dose CEC. Seventeen patients (57%) achieved a complete response. Twenty are alive (67%) and 15 (50%) are free of disease at a median follow-up time of 30 months. For 23 cycles of high-dose CEC, the median time from AuBMT to a granulocyte count > or = 0.5/microL was 11 days (range, 0 to 14) and to a platelet count 50,000/microL, 19 days (range, 14 to 34). The survival of 58 patients treated in two of our center's programs that incorporated high-dose chemotherapy (high-dose carboplatin plus etoposide [CE] and CEC) was compared with our prior experience with conventional-dose cisplatin chemotherapy alone in poor-risk GCT. Patients treated with marker-dependent, early-intervention high-dose chemotherapy experienced longer survival (P = .001). CONCLUSION: In this setting, high-dose CEC was well tolerated, cumulative toxicity was lacking, and the recovery of blood counts after AuBMT was rapid. A randomized trial has been initiated to investigate further the role of high-dose CEC in first-line therapy for patients with poor-risk GCT. PMID- 9215824 TI - Long-term follow-up of patients with good-risk germ cell tumors treated with etoposide and cisplatin. AB - PURPOSE: To assess the durability of response and overall survival for patients with good-risk metastatic germ cell tumors (GCT) treated with four cycles of etoposide and cisplatin (EP). PATIENTS AND METHODS: Two hundred fourteen patients treated with EP on two consecutive randomized trials for good-risk metastatic GCT were the subject of this retrospective study. The response to therapy, relapse and survival status, and results of salvage therapy are reported. RESULTS: One hundred ninety-five patients (91%) achieved a complete response (CR). This included 182 patients (85%) who achieved a CR to chemotherapy alone and 13 patients (6%) who achieved a CR to chemotherapy plus surgical resection of viable GCT. Seventeen patients (9%) have relapsed from CR. The median time to relapse was 10 months, and the longest duration from treatment to relapse was 36 months in a patient who received three of four planned courses of therapy. Eight patients who either achieved an incomplete response (IR) or relapsed were rendered continuously disease-free by salvage therapy and are alive. One hundred eighty-three patients (86%) are alive at a median follow-up of 7.6 years. CONCLUSION: Four cycles of EP constitute effective therapy and can be offered to patients with good-risk GCT. In patients with intermediate- and poor-risk GCT, clinical trials remain a priority to identify more effective treatment. PMID- 9215825 TI - Ifosfamide- and cisplatin-containing chemotherapy as first-line salvage therapy in germ cell tumors: response and survival. AB - PURPOSE: To evaluate the efficacy and toxicity of ifosfamide- and cisplatin containing chemotherapy as first-line salvage treatment for patients with germ cell tumors (GCT). PATIENTS AND METHODS: Fifty-six patients with advanced GCT resistant to one prior cisplatin-containing regimen were treated with a salvage chemotherapy regimen of ifosfamide, cisplatin, and either vinblastine or etoposide (VeIP/VIP). RESULTS: Twenty of 56 (36%) assessable patients achieved a complete response (CR). Thirteen (23%) are alive and continuously free of disease at a median follow-up time of 52 months; the median survival duration was 18 months. Among patients with a testis primary tumor site and a prior CR to first line therapy, 65% are alive and 41% continuously disease-free, and the median survival time has not been reached. In contrast, for patients with an extragonadal primary tumor or with a testis primary tumor site and an incomplete response (IR) to first-line therapy, 31% are alive and 15% continuously free of disease, with a median survival time of 12 months (P < .03). CONCLUSION: Ifosfamide- and cisplatin-containing therapy achieves a durable CR in a minority of patients with resistant GCT as first-line therapy. Patients with a primary testis site who relapsed from a CR to first-line cisplatin therapy have a better prognosis than patients with an extragonadal primary tumor site or an IR to first line therapy. Risk-directed clinical trials to improve response and survival in both subsets are warranted. PMID- 9215826 TI - Long-term follow-up of a phase III intergroup study of cisplatin alone or in combination with methotrexate, vinblastine, and doxorubicin in patients with metastatic urothelial carcinoma: a cooperative group study. AB - PURPOSE: A previously reported randomized intergroup trial demonstrated that combination chemotherapy with methotrexate, vinblastine, doxorubicin, and cisplatin (M-VAC) was superior to single-agent cisplatin in patients with advanced urothelial carcinoma. We conducted a long-term analysis of patients included in the intergroup trial to examine factors associated with long-term survival. PATIENTS AND METHODS: Two-hundred fifty-five assessable patients with urothelial carcinoma were randomized to receive either single-agent cisplatin (70 mg/m2 on day 1) or combination chemotherapy with methotrexate (30 mg/m2 on days 1, 15, and 22), vinblastine (3 mg/m2 on days 2, 15, and 22), doxorubicin (30 mg/m2 on day 2), and cisplatin (70 mg/m2 on day 2). Courses were repeated every 28 days. The association between patient characteristics and survival was assessed using Cox proportional hazards models. RESULTS: With long-term follow-up evaluation, survival in the M-VAC arm continues to be superior to cisplatin (P = .00015, log-rank test). Predictors of survival include performance status, histology, and the presence of liver or bone metastasis. Only 3.7% of the patients randomized to M-VAC are alive and continuously disease-free at 6 years. CONCLUSION: Long-term follow-up evaluation of the intergroup trial confirms that M-VAC is superior to single-agent cisplatin in patients with advanced urothelial carcinoma; however, durable progression-free survival is rare. Patients with non transitional-cell histology, poor performance status, and/or bone or visceral involvement fare poorly and are unlikely to benefit significantly from M-VAC chemotherapy. PMID- 9215827 TI - Correlation between thymidine phosphorylase expression and prognosis in human renal cell carcinoma. AB - PURPOSE: Thymidine phosphorylase (TP) is identical to platelet-derived endothelial cell growth factor (PD-ECGF) and has angiogenic activity. We examined whether TP expression in renal cell carcinoma (RCC) is associated with microvessel density as a marker of angiogenesis, clinicopathologic characteristics, and outcome. PATIENTS AND METHODS: The enzymatic activity and expression of TP were examined in 18 RCCs and 19 kidney tissues not grossly involved with tumor from 24 patients with 13 paired samples and 11 unpaired samples by spectrophotometry and immunoblotting. The relationship between TP expression and microvessel density was assessed by immunohistochemistry in 133 RCCs. RESULTS: The median enzymatic activity of TP in RCCs was nine fold higher than that in nonneoplastic kidney tissues (P < .001). Similar results were obtained by immunoblot analysis. According to the TP staining profile, tumors were classified as no or low, intermediate, or high TP-expressing tumors. TP positivity was significantly correlated with microvessel density. TP expression was correlated with tumor grade, but there was no significant association between TP expression and other clinicopathologic characteristics. TP expression as a prognostic variable was studied using Cox's proportional hazards model. TP overexpression was an independent prognostic factor (hazards ratio, 3.95; 95% confidence interval, 0.98 to 15.89; P = .039) as were nodal category, metastases category, tumor grade, and venous invasion. CONCLUSION: These findings suggest that TP expression is correlated with microvessel density in RCC and is an unfavorable independent prognostic factor. The future development and characterization of TP inhibitors may provide a novel approach to the therapy of RCC. PMID- 9215828 TI - Interferon alfa-2a and interleukin-2 with or without cisplatin in metastatic melanoma: a randomized trial of the European Organization for Research and Treatment of Cancer Melanoma Cooperative Group. AB - PURPOSE: The combination of interferon alfa-2a (IFN alpha) and high-dose interleukin-2 (IL-2) is active in metastatic melanoma. The addition of cisplatin (CDDP) has resulted in response rates greater than 50%. This study was performed to determine whether the addition of CDDP to a cytokine treatment regimen with IFN alpha and high-dose IL-2 influences survival of patients with metastatic melanoma. PATIENTS AND METHODS: Patients with advanced metastatic melanoma were randomly assigned to receive treatment with IFN alpha 10 x 10(6) U/m2 subcutaneously on days 1 through 5 and a high-dose intravenous decrescendo regimen of IL-2 on days 3 through 8 (18 mIU/ m2/6 hours, 18 mIU/m2/12 hours, 18 mIU/m2/24 hours, and 4.5 mIU/m2/24 hours x 3) without (arm A) or with (arm B) CDDP 100 mg/m2 on day 1. Treatment cycles were repeated every 28 days to a maximum of four cycles. RESULTS: One hundred thirty-eight patients with advanced metastatic melanoma, of whom 87% had visceral metastases, were accrued for the trial. Both regimens were feasible in a multicenter setting. The objective response rate was 18% without and 33% with CDDP (P = .04). The progression-free survival was 53 days without and 92 days with CDDP (P = .02, Wilcoxon; P = .09, log-rank). There was no statistically significant difference in survival between treatment arms, with a median overall survival duration for all patients of 9 months. CONCLUSION: The addition of CDDP to cytokine treatment with IFN alpha and IL-2 does not influence survival of patients with advanced metastatic melanoma, despite a significant increase in response rate and progression-free survival. PMID- 9215829 TI - Treatment of ocular melanoma metastatic to the liver by hepatic arterial chemotherapy. AB - PURPOSE: Ocular melanoma is characterized by a high rate of liver metastases and is associated with a median survival time less than 5 months. There is no standard treatment available. Treatment strategies have, without success, relied on the experience with metastatic cutaneous melanoma. The only effective treatment is chemoembolization using cisplatin and polyvinyl sponge, which has never become accepted on a large scale. The objective of the study was to establish prospectively the efficacy and toxicity of hepatic intraarterial fotemustine, a third-generation nitrosourea, in patients with liver metastases from ocular melanoma. PATIENTS AND METHODS: Thirty-one patients were subjected to laparotomy to place a totally implantable catheter into the hepatic artery and received fotemustine 100 mg/m2 as a 4-hour infusion, first once a week for four times and then, after a 5-week rest period, every 3 weeks until progression or toxicity. Cox regression models were used to assess the prognostic role of patient survival characteristics. RESULTS: Objective responses were observed in 12 of 30 assessable patients (40%; 95% confidence interval, 22% to 59%). The median duration of response was 11 months and the median overall survival time, 14 months. Lactate dehydrogenase (LDH) appeared to be the strongest prognostic factor for survival. Toxicity was minimal and treatment could be administered on an outpatient basis. CONCLUSION: The results of hepatic arterial chemotherapy with fotemustine produced a high response rate and survival similar to chemoembolization therapy. It involves no major toxicity and preserves the quality of life. To assess further its effectiveness, a randomized study to compare hepatic intraarterial versus intravenous chemotherapy is being planned. PMID- 9215830 TI - Phase II study of continuous infusion carmustine and cisplatin followed by cranial irradiation in adults with newly diagnosed high-grade astrocytoma. AB - PURPOSE: To evaluate the activity and toxicity of carmustine (BCNU) and cisplatin administered as a 72-hour continuous intravenous infusion before radiation in adults with newly diagnosed high-grade astrocytomas. PATIENTS AND METHODS: Fifty two patients with a Karnofsky performance status greater than 60 and no prior antineoplastic therapy entered this protocol. The median age of the patients was 55 years. Eighty-eight percent had glioblastoma multiforme and 12% had anaplastic astrocytomas. BCNU (40 mg/m2/d) and cisplatin (40 mg/m2/d) were administered concurrently as a 72-hour infusion every 3 to 4 weeks. Radiation was begun 4 weeks after the third cycle of chemotherapy or earlier for progressive disease. Responses required a > or = 50% reduction in contrast-enhancing volume. RESULTS: Forty patients (77%) completed three chemotherapy infusions, five (10%) received two infusions, and seven (13%) received only one. Fifty-one patients completed radiation. Seventeen (42%) patients with measurable disease had a partial response (PR) to chemotherapy, 23 (53%) had stable disease (SD), and two (4%) had progressive disease (PD) on chemotherapy. The median survival time for all patients was 13 months. Survival rates at 1, 2, 3, and 5 years were 62%, 19%, 12%, and 5%, respectively. Grade III to IV leukopenia occurred in 32% of patients; 63% received platelet transfusions and 58% required RBCs. Neutropenic fevers were rare and no intracranial hemorrhages or treatment-related deaths were noted. Nausea, vomiting, peripheral neuropathy, hearing loss, and thromboembolic events were relatively common. CONCLUSION: This chemotherapy regimen appears to have significant activity and may prolong survival in adults with newly diagnosed high-grade astrocytoma. PMID- 9215831 TI - Tumoral-reduced folates and clinical resistance to fluorouracil-based treatment in head and neck cancer patients. AB - PURPOSE: To describe the distribution of tumoral-reduced folates in cancer patients and to analyze the link between this parameter and antitumor efficacy of fluorouracil (FU)-based induction chemotherapy. PATIENTS AND METHODS: Ninety-five patients with head and neck squamous cell carcinoma were included in the present study and 41 received induction treatment with FU-based chemotherapy (35 men and six women; mean age, 59 years; range, 40 to 76). Thymidylate synthase (TS) activity was measured according to the tritium-release assay. Reduced folates (5 10 methylenetetrahydrofolate [CH2FH4] plus tetrahydrofolate) were measured according to the entrapment assay. RESULTS: Among the whole group of patients, reduced folates ranged from nondetectable (< 0.3) to 17.7 pmol/mg protein; CH2FH4 ranged from nondetectable (< 0.3) to 8.2 pmol/mg protein. There was no significant link between tumoral levels of reduced folates and the severity of disease stage. Among 41 treated patients, there were 12 (29%) complete responses (CRs), 18 (44%) partial responses (PRs), and 11 patients (27%) with no response (NR). No statistically significant relationship was observable between TS activity and response. The distribution of CH2FH4 in tumors was significantly higher for complete responders in comparison to patients with a PR or NR. CONCLUSION: The present clinical data demonstrate the importance of basal tumoral reduced folates for the achievement of optimal efficacy of FU-based treatment. They may have implications for the identification of patients resistant to FU chemotherapy and for a better understanding and management of FU modulation by folinic acid (FA). PMID- 9215832 TI - Ewing's sarcoma and primitive neuroectodermal tumor in adults: are they different from Ewing's sarcoma and primitive neuroectodermal tumor in children? AB - PURPOSE: To determine whether age at diagnosis influences the behavior of Ewing's sarcoma and primitive neuroectodermal tumor (PNET). PATIENTS AND METHODS: We reviewed the clinical features, treatment, and outcome of 59 consecutive patients with Ewing's sarcoma and PNET treated on the Adult Sarcoma Unit at our institution from 1980 to 1995. RESULTS: The 37 male and 22 female patients had a median age of 24 years. Lower limb was the most common primary tumor site. Fifteen patients had nonmetastatic tumor less than 100-mL volume, 27 had nonmetastatic disease greater than 100-mL volume, and 17 had evidence of metastatic disease at presentation. The origin of the primary tumor was soft tissue in 28 cases, bone in 30, and uncertain in one. The Kaplan-Meier estimate of 5-year overall survival (OS) in all patients was 38% and of progression-free survival (PFS), 27%. When patients with metastatic disease at presentation were excluded, these figures increased to 52% and 34%, respectively. Bulk of disease at presentation and response to primary treatment were statistically highly significant predictors of both PFS and OS. Age and tissue of origin of the tumor did not influence outcome. CONCLUSION: The behavior of Ewing's sarcoma and PNET in adults is no different from its behavior in children. We feel the way forward in the treatment of adults with Ewing's sarcoma and PNET is for them to be included in the current multicenter trials of multidisciplinary treatment directed at children. PMID- 9215833 TI - Does histology influence outcome in childhood Hodgkin's disease? Results from the United Kingdom Children's Cancer Study Group. AB - PURPOSE: Histology has been identified as an important prognostic factor in Hodgkin's disease (HD) in adults. Information regarding the impact of histology on outcome in childhood HD is scarce. This study determines the effect of histology on the overall survival (OS) or progression-free survival (PFS) in a national series of children treated in a standardized manner. PATIENTS AND METHODS: The results of treatment of 331 assessable patients, treated between January 1, 1982 and June 30, 1992, in the United Kingdom Children's Cancer Study Group (UKCCSG) Hodgkin's study I were reviewed to evaluate OS, PFS, and deaths according to stage and histology. Treatment was either involved-field radiation alone (stage IA) or chlorambucil, vinblastine, procarbazine, and prednisolone (ChlVPP) chemotherapy with or without mediastinal radiation. All were clinically staged at diagnosis. RESULTS: Nodular sclerosing (NS) HD was the most common histologic subtype (155 of 331 patients [47%]) and was uniformly distributed through all stages. Lymphocyte-depletion (LD) HD was extremely uncommon (< 1%). Mixed-cellularity (MC) HD had the highest relapse rate, but this was only significant (P < .05) in stage I patients who received local irradiation alone. There was no other statistically significant difference in OS and PFS between the various histologic subtypes. Multivariate analysis for PFS and OS confirmed that stage was the most important prognostic factor and that histology did not have an effect after stratification by stage. CONCLUSION: This study demonstrates that with effective multiagent chemotherapy, histologic subtype does not influence outcome. The high relapse rates in stage I MC subtype indicates that MC HD is biologically aggressive and systemic treatment with or without local irradiation may be indicated. The high relapse rate in stage IV patients appeared to be independent of histology. PMID- 9215834 TI - Use of chest computed tomography in the staging of pediatric Wilms' tumor: interobserver variability and prognostic significance. AB - PURPOSE: To determine the specificity and prognostic significance of computed tomography (CT) of the chest in pediatric Wilms' tumor. PATIENTS AND METHODS: Patients treated for newly diagnosed Wilms' tumor at St Jude Children's Research Hospital between December 1978 and July 1995 were included in the study if an initial chest radiograph and CT were available and if pulmonary involvement (determined by chest radiographs) was absent. For the 202 patients studied, radiographs and CT scans were reviewed blindly and independently by three experienced radiologists for the presence of pulmonary nodules. Outcome variables consisted of intraobserver variability (in a subsample of 40 cases) and concordance between ratings on radiographs and CT scans (both by McNemar's test), interrater variability (by logistic regression), and the cumulative incidence of pulmonary relapse for patients with and without positive CT scans, by reviewer. RESULTS: As expected, ratings of pulmonary involvement on radiographs were discordant with CT ratings. There was marked variability among reviewers in CT ratings (P = .0001). Of 202 CT scans, 78 were read as positive by at least one reviewer, 41 were rated positive by only one reviewer, 18 by two reviewers, and 19 by all three. Intrarater variability on repeat reviews was not significant. Patients with nodules identified on CT had a significantly higher pulmonary relapse rate when analyzed separately by reviewer. However, for the 14 patients who had pulmonary relapse, CT scans were rated positive by all three reviewers in only five cases and as negative by all three in another five cases. CONCLUSION: The variability in interpretation of chest CT scans in patients with Wilms' tumor limits the predictive utility of these studies. Optimal, standardized techniques and central review are essential if chest CT is to be used for staging in cooperative studies. PMID- 9215836 TI - Access to bone marrow transplantation for leukemia and lymphoma: the role of sociodemographic factors. AB - PURPOSE: Use of bone marrow transplantation (BMT), a complex, costly treatment for many forms of cancers, has increased significantly in recent years. The increasingly competitive health care marketplace raises concerns about patient access to costly medical procedures such as BMT. We attempted to evaluate patient access to BMT for the treatment of leukemias and lymphomas. METHODS: We analyzed inpatient hospital discharge data from four states (California, Maryland, Massachusetts, and New York) for 2 years (1988 and 1991) to examine whether the use of BMT for patients with either leukemia or lymphoma varies by sociodemographic characteristics and insurance coverage. We developed a sorting algorithm to collapse the discharge data into patient level records. We used logistic regression to analyze the odds of receiving a BMT stratified by disease type (leukemia or lymphoma). RESULTS: After controlling for other factors, black patients with leukemia are 51% to 53% as likely as whites, while black patients with lymphoma are 34% to 45% as likely as white patients to undergo a BMT (P < .05). Medicaid, self-pay patients, and Health Maintenance Organization (HMO) enrollees with either leukemia or lymphoma are significantly less likely to undergo a BMT compared with patients with private insurance. Younger patients are significantly more predisposed to undergo a BMT than older patients. The odds of receiving a BMT have increased over time, but the rates of increase vary by state. Consistent with clinical expectations, the relative odds of BMT vary significantly by type of leukemia or lymphoma. CONCLUSION: Substantial variation exists in access to BMT for patients with either leukemia or lymphoma. Black patients, those enrolled in HMOs, those covered by Medicaid, and self-pay patients were less likely to receive a BMT when admitted for either leukemia or lymphoma. These findings raise concerns about access to cancer treatments for patients in the current health care system. PMID- 9215835 TI - EMA/CO for high-risk gestational trophoblastic tumors: results from a cohort of 272 patients. AB - PURPOSE: To evaluate the results of etoposide, methotrexate, and dactinomycin alternating with cyclophosphamide and vincristine (EMA/CO) chemotherapy in women with high-risk gestational trophoblastic tumors (GTT) and to document the middle- and long-term toxicity of the regimen. PATIENTS AND METHODS: A total of 272 consecutive women with high-risk GTT, including 121 previously treated patients, were treated with weekly EMA/CO. The median follow-up duration is 4.5 years (range, 1 to 16). RESULTS: The cumulative 5-year survival rate is 86.2% (95% confidence interval, 81.9% to 90.5%). No deaths from GTT have occurred later than 2 years after the end [corrected] of EMA/CO. In a multivariate model, adverse prognostic factors were the presence of liver metastases (P < .0001), interval from antecedent pregnancy (P < .0001), brain metastases (P = .0008), and term delivery of antecedent pregnancy (P = .045). There were 11 (4%) early deaths, while 213 patients (78%) achieved a complete remission. Forty-seven (17%) developed drug resistance to EMA/CO, of whom 33 (70%) were salvaged by further cisplatin-based chemotherapy and surgery. Two women developed acute myeloid leukemia, two cervical malignancy, and one gastric adenocarcinoma after EMA/CO. More than half (56%) of the women who had fertility-conserving surgery and who have been in remission at least 2 years have become pregnant since the completion of EMA/CO, with 112 live births, including three infants with congenital abnormalities. CONCLUSION: EMA/CO is an effective and well-tolerated regimen for high-risk GTT. More than half of the women will retain their fertility; however, there is a small but significant risk of second malignancy. PMID- 9215837 TI - Randomized trial of CVPP for three versus six cycles in favorable-prognosis and CVPP versus AOPE plus radiotherapy in intermediate-prognosis untreated Hodgkin's disease. AB - PURPOSE: To evaluate in a randomized trial the impact of three versus six cycles of cyclophosphamide, vinblastine, procarbazine, and prednisone (CVPP) chemotherapy in favorable-prognosis and CVPP versus doxorubicin, vincristine, prednisone, and etoposide (AOPE) plus involved-field radiotherapy (RT) in intermediate-prognosis previously untreated Hodgkin's disease. PATIENTS AND METHODS: Of 256 patients evaluated, 80 with a favorable prognosis according to a prognostic index designed by the Grupo Argentina de Tratamiento de Leucemia Aguda (GATLA) were randomized to three versus six cycles of CVPP without RT and 176 with intermediate risk to CVPP versus AOPE, both for six cycles with RT between the third and fourth cycles of 30 Gy to the involved areas at diagnosis. CVPP consisted of intravenous (I.V.) cyclophosphamide and vinblastine on days 1 and 8, and oral procarbazine and prednisone on days 1 to 14, every 28 days. AOPE consisted of I.V. doxorubicin and vincristine on day 1, oral prednisone on days 1 to 5, and I.V. etoposide on days 1 and 3, every 28 days. RESULTS: Complete remission was obtained in 39 of 41 (95%) patients treated with three cycles of CVPP and 36 of 39 (92%) treated with six cycles in the favorable-risk group (difference not significant [NS]). In the intermediate-risk group, 89 of 92 (97%) treated with CVPP plus RT versus 75 of 84 (89%) treated with AOPE plus RT achieved a complete remission (P = .05). At 60 months, the event-free survival (EFS) and overall survival rates in the favorable-risk group were 80% and 91% for CVPP x 3 and 84% and 97% for CVPP x 6, respectively (P = NS). In the intermediate risk group, 60-month EFS rate for CVPP plus RT was 85%, compared with 66% for AOPE plus RT (P = .009). The overall survival rate was 95% versus 87% respectively (P = .157). CONCLUSION: Three cycles of CVPP without RT are equally effective as six cycles in the favorable-risk group. However, in the intermediate group, CVPP plus RT is superior to AOPE plus RT, with significantly fewer events before and after induction (P = .009), without a difference in overall survival. PMID- 9215838 TI - Unique role of cytogenetics in the prognosis of patients with myeloma receiving high-dose therapy and autotransplants. AB - PURPOSE: Although important predictors of survival in myeloma patients have been identified, it is well recognized that better prognostic factors for this disease are needed. Because cytogenetics play a dominant role in the outcome of patients with acute leukemia, their prognostic value was evaluated in a large group of newly diagnosed and previously treated myeloma patients receiving autotransplants. METHODS: A total of 427 either newly diagnosed (26%) or previously treated patients (74%) received tandem transplants, supported by mobilized peripheral-blood stem cells. Numerous variables, including cytogenetics, were analyzed for their impact on complete remission, event-free survival (EFS), and overall survival (OS). RESULTS: Abnormal karyotypes were detected in 37% of our patients and were very complex, irrespective of the duration of standard therapy before the first autotransplant. In addition to previously recognized unfavorable implications of partial or complete deletion of chromosome 13 and 11q abnormalities, we now observed that the presence of any translocation likewise portended poor outcome (unfavorable karyotypes). On multivariate analysis, the absence of an unfavorable karyotype was the most favorable variable for both EFS (P = .0001) and OS (P = .0001). Other favorable factors were duration of standard therapy and a low beta-2 microglobulin (B2M) level before the first autotransplant. A risk-based classification system was developed according to the number of these favorable variables present, showing highly significant differences in event-free and overall survival. CONCLUSION: Cytogenetics play a dominant role in myeloma and were independent of previously recognized important prognostic factors, such as B2M and duration of prior standard therapy. PMID- 9215839 TI - Treatment of T-cell prolymphocytic leukemia with human CD52 antibody. AB - PURPOSE: T-prolymphocytic leukemia (T-PLL) is an aggressive malignancy of mature T cells refractory to conventional chemotherapy, with a median survival duration of 7.5 months. We report here promising results with the use of a genetically reshaped human CD52 antibody, CAMPATH-1H. PATIENTS AND METHODS: Fifteen patients with T-PLL, most of whom had received the purine analog deoxycoformycin (DCF), were treated with CAMPATH-1H. Results were compared with those of 25 patients treated with DCF. RESULTS: Major responses occurred in 11 patients (73%) treated with CAMPATH-1H compared with 40% with DCF. Complete remissions (CRs) were documented in nine (60%) of the CAMPATH-1H cases and only three (12%) were obtained with DCF. CRs with CAMPATH-1H were durable, and re-treatment with the antibody resulted in second CRs in three relapsed patients. Two of them were successfully autografted with peripheral-blood and bone marrow stem cells collected during the first CR. Apart from first-dose reactions, infusions of CAMPATH-1H were well tolerated. However, two responding patients developed severe bone marrow aplasia that was fatal in one; the second remained moderately pancytopenic 21 weeks after stopping CAMPATH-1H therapy. The cause of this adverse effect is unknown. CONCLUSION: CAMPATH-1H is an effective agent in T-PLL and represents a significant improvement over other types of therapy. However, CAMPATH-1H alone is not sufficient for long-term remissions, and the role of autologous stem-cell transplantation needs further investigation. PMID- 9215840 TI - Cost-effectiveness of interferon alfa in chronic myelogenous leukemia. AB - PURPOSE: To evaluate the cost-effectiveness of interferon alfa (IFN alpha) treatment of patients with chronic myelogenous leukemia relative to conventional chemotherapy. MATERIALS AND METHODS: A decision-analysis model that involved a multistate Markov process was designed to estimate the expected cost and quality adjusted life expectancies for two cohorts of patients to be administered conventional chemotherapy or IFN alpha. Two IFN alpha strategies were modeled: prolonged treatment for patients who achieved a hematologic response (scenario A) or only for patients who achieved a cytogenetic remission in a 2-year period (scenario B). Data on response and transition probabilities between health states were obtained from the literature by a MEDLINE search and pooled with a meta analytic method. Costs were based on local charges. Expected survival was adjusted for quality of life on the basis of an expert panel judgment. RESULTS: Baseline analysis showed IFN alpha treatment to increase the quality-adjusted life expectancy by 15.5 and 12.5 months relative to conventional chemotherapy, in scenarios A and B, respectively. Marginal cost-effectiveness was $89,500 and $63,500 per quality-adjusted life-year (QALY) gained. Sensitivity analysis confirmed IFN alpha as the most effective approach. Cost-effectiveness results were sensitive to the cost of IFN alpha therapy and to the assumptions about the rate of cytogenetic remission. Reducing the drug dose, as suggested by a recent report, would decrease the marginal cost-effectiveness to less than $20,000. CONCLUSION: IFN alpha is substantially superior to conventional chemotherapy in terms of quality-adjusted survival, but, at the current doses, marginal cost effectiveness ranges from $50,000 to $100,000 per QALY gained under most of our assumptions. PMID- 9215841 TI - Detection of hepatocellular carcinoma with a new alpha-fetoprotein antibody imaging kit. AB - PURPOSE: The aim of this phase II study was to assess the clinical utility and safety of AFP-Scan (Immunomedics, Inc, Morris Plains, NJ), a technetium-99m (99mTc)-labeled anti-alpha-fetoprotein (AFP) Fab' imaging kit, in the evaluation of hepatocellular carcinoma (HCC), in comparison to computed tomography (CT). PATIENTS AND METHODS: Twenty-five consecutive patients with a history of HCC were examined by planar and single-photon emission computed tomography (SPECT) imaging at 6 and 24 hours after intravenous (I.V.) injection of 1 mg AFP-Scan labeled with 925 MBq 99mTc. RESULTS: In 20 patients, there was a specific binding of the Fab' antibody to the tumor, whereas in four patients who presented with elevated serum AFP levels, no specific targeting was found and no malignant lesions were evident by CT or biopsy. One patient was diagnosed as false-negative by AFP-Scan. In five of six patients with normal serum AFP levels, focal uptake was demonstrated. In one case, metastatic disease in the lower abdomen was found. In all patients, diagnostically relevant information was provided by the 24-hour antibody images, especially with SPECT. Comparing AFP-Scan versus CT, the former showed a higher sensitivity (95% v 63%) and specificity (67% v 17%), with an overall accuracy of 88% versus 52% for AFP-Scan versus CT, even in patients with normal serum AFP titers. No adverse reactions or human antimouse antibody (HAMA) elevations were found in any of the patients. CONCLUSIONS: AFP-Scan appears to be a promising new antibody imaging kit for the disclosure of sites of HCC and should aid in the management of these patients by revealing primary, recurrent, and metastatic disease with a single imaging modality. PMID- 9215842 TI - Phase I and pharmacokinetic study of an oral platinum complex given daily for 5 days in patients with cancer. AB - PURPOSE: We aimed to determine the maximum-tolerated dose (MTD) clinical toxicities, pharmacokinetics, and pharmacodynamics of oral JM216 given once daily for 5 days to cancer patients. PATIENTS AND METHODS: Patients who fulfilled standard phase I trial criteria were enrolled. Oral JM216 was given at doses based on patient body-surface area, on an empty stomach, once daily for 5 consecutive days, as 10-, 50-, and 200-mg hard gelatin capsules and with oral antiemetics. The pharmacokinetics of platinum were studied on days 1 and 5 of the first treatment course using atomic absorption spectrophotometry (AAS). RESULTS: Thirty-two patients received 94 courses of oral JM216 at doses that ranged from 30 to 140 mg/m2 body-surface area for 5 consecutive days. The MTD was 140 mg/m2/d. The dose-limiting toxicities were thrombocytopenia and neutropenia. Hematotoxicity was reversible (nadir, 17 to 21 days; recovery, 28 days), noncumulative, and dependent on the dose and history of previous therapy. There were two instances of neutropenic sepsis. Two-thirds of patients experienced mild nausea, vomiting, or diarrhea. There was no ototoxicity, neurotoxicity, nephrotoxicity, or objective tumor responses. There was a significant correlation between JM216 dose and the day 1 and 5 plasma ultrafiltrate area under the concentration-time curve (AUC; r = .78), which indicates linear pharmacokinetics. There was considerable intersubject pharmacokinetic and pharmacodynamic variability, but a significant sigmoidal relationship between the plasma ultrafiltrate AUC and severity of thrombocytopenia (R2 = .83). CONCLUSION: We recommend JM216 doses of 100 and 120 mg/m2/d x 5 for previously treated and untreated patients, respectively, for phase II trials. PMID- 9215843 TI - Limitations of reverse-transcriptase polymerase chain reaction analyses for detection of micrometastatic epithelial cancer cells in bone marrow. AB - PURPOSE: This study was designed to evaluate the potential of reverse transcriptase polymerase chain reaction (RT-PCR) analyses for the detection of micrometastatic carcinoma cells in bone marrow (BM). PATIENTS AND METHODS: The specificity of RT-PCR assays with primers specific for various tumor-associated and organ-specific mRNA species was examined by analysis of 53 BM aspirates from control patients with no epithelial malignancy. In addition, BM samples from 63 patients with prostate cancer (n = 53) or breast cancer (n = 10) were analyzed by RT-PCR with primers specific for prostate-specific antigen (PSA) mRNA. As a reference method, all samples were analyzed simultaneously by an established immunocytochemical assay, using monoclonal antibodies (mAbs) against cytokeratins (CK) for tumor-cell detection. RESULTS: Seven of eight marker species could be detected in a considerable number of BM samples from control patients: epithelial glycoprotein-40 (EGP-40; 53 of 53 samples), desmoplakin I (DPI I; five of five), carcinoembryonic antigen (CEA; five of 19), erb-B2 (five of seven), erb-B3 (six of seven), prostate-specific membrane antigen (PSM; four of nine), and CK18 (five of seven). Only PSA mRNA was not detected in any of the 53 control BM samples. In serial dilution experiments, the PSA RT-PCR assay was able to detect five LNCaP prostate carcinoma cells in 4 x 10(6) BM cells. CK-positive cells were found in 20 patients (37.7%) with prostate cancer, while PSA mRNA was found in only 15 (28.3%; P = .04). Moreover, despite the recent observation that PSA is also expressed in mammary carcinomas, none of the 10 CK-positive BM samples were PSA mRNA-positive. CONCLUSION: Limiting factors in the detection of micrometastatic tumor cells by RT-PCR are (1) the illegitimate transcription of tumor-associated or epithelial-specific genes in hematopoietic cells, and (2) the deficient expression of the marker gene in micrometastatic tumor cells. PMID- 9215844 TI - Low-dose urokinase infusions to treat fibrinous obstruction of venous access devices in cancer patients. AB - PURPOSE: This study was undertaken to determine the role of low-dose urokinase infusions in treating fibrinous occlusions of venous access devices (VADs) in cancer patients. PATIENTS AND METHODS: Forty-two patients with VAD occlusions refractory to routine urokinase instillations were documented by x-ray (cathetergram) to have fibrin sleeves at the catheter tips. They were randomized to receive infusions of either urokinase (40,000 U/h) or urokinase with heparin (320 U/h) through their catheters. After 1, 3, 6, and 12 hours of treatment, the function of the VADs was reassessed. Whenever the obstruction had been relieved, the infusion was stopped and a repeat cathetergram was performed. The status of the unoccluded catheters was followed to determine the longevity of the restored function. RESULTS: Twenty-one catheters were treated with urokinase alone and 21 with the combination of urokinase and heparin. In each group, 16 VADs opened within 12 hours of treatment and five did not. By actuarial analysis, the probability was only 0.28 that a reopened catheter would reocclude within 6 months. CONCLUSION: Low-dose urokinase infusions can restore function to the majority of catheters occluded by fibrin sleeves. Adding heparin to the urokinase does not enhance the efficacy of the infusions. The restored function often persists until the VADs are removed. PMID- 9215846 TI - Are there sex biases in standardized tests of radiation oncology knowledge? AB - PURPOSE/OBJECTIVE: Recent studies have identified biases directed against women in standardized tests. We tested for the existence of such biases in the American College of Radiology (ACR) In-Training Examination in Radiation Oncology and the American Board of Radiology (ABR) Written Radiation Oncology Board Examination. MATERIALS AND METHODS: Our request to the ABR to permit us to study performance on their examinations, as a function of sex, was refused. We obtained scores, through the cooperation of six academic radiation oncology departments, for residents-in-training taking the in-service examination and candidates taking the written board examination for the first time. Test results for 1984 to 1995 were blinded as to name, but not sex or institution of training. For the in-service examination, scores are reported as percentiles normalized to the year of training. The effect of multiple scores for the same resident was assessed using a repeated-measures analysis of variance. Residents were nested within each sex/institution combination and crossed with training year and calendar year. The effects of three factors (sex, institution, and year the examination was taken) on the results of the biology, physics, and clinical sections were evaluated with an analysis of variance. The interactions of sex with institution and year were included to determine the scope of the sex effect. For the board examination, scores are reported as percentiles, as well as an overall pass/ fail outcome. An analyses of variance was performed similar to that used for the in-service examination. In addition, Fisher's exact test and logistic regression were used to analyze overall outcome (pass/fail). RESULTS: We obtained data for 79 residents (48 men and 31 women, 1.54:1) who took the in-service examinations 165 times. Sixty-two residents (41 men and 21 women, 1.95:1) had an initial sitting for the ABR written examination. On the in-service examination, for the biology, physics, and clinical subsections, calendar year, training year, and sex did not have a significant effect on examinees scores. Institution of training had a significant effect (P < .02) on the scores in biology and physics. The total in service examination scores were not significantly influenced by calendar year, training year, or sex. Institution of training has a strong influence on overall score (P = .03) and the interaction of sex with training year is near significance level (P = .06). The power for our statistical tests ranged from 0.88 to 0.99. On the board examination, sex, institution of training, year the examination was taken, and interaction of sex with year or sex with institution of training did not have a significant effect on test scores. Pass rates were 90% for men versus 81% for women (P = .43). CONCLUSION: Sex did not significantly influence the results of the in-service examination or the written board examination. Institution of training is the strongest influence on the results of the in-service examination. PMID- 9215845 TI - Randomized phase III trial evaluating the role of erythropoietin in the prevention of chemotherapy-induced anemia. AB - PURPOSE: Although erythropoietin (EPO) is known to be useful in treating chemotherapy-induced anemia, few data are available on its potential preventive role. The aim of this study was to evaluate the ability of EPO in preventing the development of clinically significant anemia in patients treated with chemotherapy. PATIENTS AND METHODS: Sixty-two early-stage breast cancer patients undergoing accelerated adjuvant chemotherapy were randomized to receive EPO 150 U/kg three times a week or no additional treatment. Chemotherapy consisted of six cycles of cyclophosphamide 600 mg/m2, epirubicin 60 mg/m2, and fluorouracil 600 mg/m2 (CEF) intravenously on day 1, every 2 weeks with the support of granulocyte colony-stimulating factor (G-CSF), 5 microg/kg subcutaneously from day 4 to day 11. RESULTS: Throughout the six cycles of chemotherapy, EPO-treated patients maintained stable values of hemoglobin, whereas control patients developed a progressive anemia. At the end of chemotherapy, the mean (+/- SD) hemoglobin decrease in the control group was 3.05 g/dL (+/- 1.0; 95% confidence interval [CI], 2.6 to 3.5), whereas in the EPO group it was 0.8 (+/- 1.4; 95% CI, 0.3 to 1.4). Clinically significant anemia (hemoglobin < or = 10 g/dL) occurred in 16 patients (52%; 95% CI, 33 to 69) in the control arm and in no patient (0%; 95% CI, 0 to 14) in the EPO arm (P = .00001). CONCLUSION: EPO prevents anemia in patients undergoing chemotherapy. Further trials are required to identify subsets of patients in which the preventive use of this drug could be cost-effective. PMID- 9215847 TI - Irradiation in the setting of collagen vascular disease: acute and late complications. AB - PURPOSE: Based on reports of greater toxicity from radiation therapy, collagen vascular diseases (CVDs) have been considered a contraindication to irradiation. We assessed the complications of radiation therapy in patients with CVD. PATIENTS AND METHODS: A total of 209 patients with documented CVD were irradiated between 1960 and 1995. One hundred thirty-one had rheumatoid arthritis (RA), 25 had systemic lupus erythematosus (SLE); 17 had polymyositis or dermatomyositis; 16 had scleroderma; eight had ankylosing spondylitis; five had juvenile RA; three had discoid lupus erythematosus; and four had 4 mixed connective tissue disorders (MCTD). The mean follow-up duration of curative cases was more than 6 years. Doses ranged from 10 to 87.6 Gy, with a median of 45 Gy. RESULTS: Overall, 263 sites were assessable in 209 patients. Significant (> or = grade 3) acute toxicity was seen in 10% of irradiated sites. Severe late effects were associated with significant acute reactions and with non-RA CVDs (6% v 21% at 5 years). No difference was seen in late effects according to timing of CVD onset, presence of concurrent vascular insults, radiation dose, or other technical factors, or by measures of disease activity. CONCLUSION: RA does not appear to have an elevated rate of late toxicity. While non-RA CVD is significantly associated with increased radiation late effects at standard doses, radiation-related mortality remains exceedingly rare. The choice of therapeutic modality in this radiosensitive group of patients should be made on a case-by-case basis. PMID- 9215848 TI - Role of independent data-monitoring committees in randomized clinical trials sponsored by the National Cancer Institute. AB - PURPOSE: To describe the rationale for independent data monitoring committees (DMCs) for National Cancer Institute (NCI)-sponsored phase III cooperative group clinical trials. DESIGN: We review the necessity for interim monitoring of outcome data during the course of randomized clinical trials and summarize the reasons for establishing DMCs with requisite expertise and with appropriate independence from study investigators. RESULTS: The important components of the policy for cooperative group DMCs are described with a focus on the makeup of these bodies and on the complementary roles of study committee leadership and DMCs in protecting patient safety during the conduct of randomized clinical trials. CONCLUSION: The cooperative group DMCs that are independent of the study committees and that have the requisite expertise to examine accumulating data and to base decisions on monitoring guidelines that are specified in advance by the study committee provide a body able to protect patient safety, to protect the integrity of the clinical experiments on which patients have consented to participate, and to assure the public that conflicts of interest do not compromise either patient safety or trial integrity. PMID- 9215849 TI - Brain tumor-polyposis syndrome: two genetic diseases? AB - PURPOSE AND DESIGN: This report presents a comprehensive and statistical analysis of the brain tumor-polyposis (BTP) cases referred to as Turcot's syndrome in the literature. RESULTS: BTP patients encompass a heterogeneous group that can be classified into two statistically distinct clinical entities based on phenotype of the polyps (P = .0001), presence of colorectal cancer (P = .0001), type of brain neoplasm, ie, glioma or medulloblastoma (P = .0001), presence of skin lesions (P = .0004) and cafe-au-lait spots (P = .0008), as well as consanguinity (P = .0135). CONCLUSION: The first entity (BTP syndrome type 1) consists of patients who have glioma and colorectal adenomas without polyposis (non-FAP cases), and their siblings with glioma and/or colorectal adenomas. For these patients, we show that the patient's age at malignant glioma occurrence is less than 20 years (50 to 80 years in the general population), which strongly supports the existence of an underlying genetic cause. The neoplasms of these patients show DNA replication errors, which suggests a relationship with hereditary nonpolyposis colorectal cancer (HNPCC), a disease characterized by germline alterations in DNA mismatch repair genes. The second entity (BTP syndrome type 2) consists of patients with a CNS tumor that occurs in a familial adenomatous polyposis kindred (FAP cases). These patients carry germline mutations in the APC gene, which suggests that mutations in this gene might predispose to brain tumors. Risk analysis shows increased incidence of medulloblastoma in FAP patients, but APC mutations are not found in sporadic glioma or medulloblastoma. Therefore, further investigations should establish whether the occurrence of medulloblastoma in an FAP family represents a variant of FAP. PMID- 9215850 TI - Hypertrophic osteoarthropathy. PMID- 9215851 TI - Low-grade glioma--the Europeans are winning! PMID- 9215852 TI - Claims-based 'report card' deserves failing grade. PMID- 9215853 TI - Trial etiquette. PMID- 9215854 TI - The growth inhibitory effect of conjugated linoleic acid on MCF-7 cells is related to estrogen response system. AB - Conjugated linoleic acid (CLA) has been shown to have a direct oncostatic action on MCF-7 human breast cancer cells in culture. However, the mechanism involved is not fully elucidated. In this study we have examined whether the inhibitor action is related to the estrogen responsiveness of MCF-7 cells. Our results demonstrate that CLA selectively inhibits proliferation of ER positive MCF-7 cells as compared with ER negative MDA-MB-231 cells. Cell cycle studies indicated that a higher percentage of CLA treated MCF-7 cells remained in the G0/G1 phase as compared to control and those treated with linoleic acid (LA). CLA also inhibited expression of c-myc in MCF-7 cells. These results demonstrate that CLA may inhibit MCF-7 cell growth by interfering with the hormone regulated mitogenic pathway. We are reporting for the first time the involvement of CLA, a dietary factor, in the regulation of hormone mediated mitogenic pathways in ER positive breast cancer cell proliferation in vitro. PMID- 9215855 TI - Effect of antiproliferative flavonoids on ascorbic acid accumulation in human colon adenocarcinoma cells. AB - Dietary flavonoids were found to be antiproliferative for human colon cancer cells, Caco-2 and HT-29, and rat nontransformed intestinal crypt cells, IEC-6. The antiproliferative potency was found to be structure-dependent. We report here a correlation between the antiproliferative potency of these flavonoids and their ability to inhibit cellular accumulation of ascorbic acid (vitamin C). Caco-2, HT 29 and IEC-6 cells were found to accumulate ascorbic acid in a sodium-dependent fashion although some ascorbic acid may also enter the cells through sodium independent mechanisms. Flavonoids that have been found to be antiproliferative, quercetin and genistein, inhibited the accumulation of ascorbic acid. The inhibition was dose-dependent and could be observed after as short as 10-min of incubation. The degree of inhibition of accumulation was more during rapid cell division as compared to post-confluency Caco-2 cells. Flavonoids that were found to show little antiproliferative effect, naringenin and catechin, also had little effect on ascorbic acid accumulation. The antiproliferative property of flavonoids could be linked to their ascorbic acid deprivation property. PMID- 9215856 TI - Rapid cytocidal and cytostatic chemosensitivity test by measuring total amount of mRNA. AB - Chemosensitivity of vinblastin, cisplatin, and mitomycin C were assessed in four different human cancer cell lines (U937, HL-60, CaR-1, and HepG2) by MTT (3-(4,5 dimethylthiazol-2-yl)-2.5-diphenyltetrazolium bromide) assay and the measurement of total cytosolic poly(A) + mRNA. Results of 12 h mRNA assay with 10 x peak plasma concentration (PPC) were significantly correlated with that of standard 3 days MTT assay with 1 x PPC. Furthermore, mRNA assay was changed more significantly than MTT assay under cytostatic condition. Because of its minimum culture requirement (12 h) and broad spectrum (cytocidal and cytostatic), mRNA assay will become a useful tool for chemosensitivity test. PMID- 9215857 TI - Tumor cell growth is inhibited by suppressing metallothionein-I synthesis. AB - The effect of metallothionein (MT)-I antisense oligodeoxynucleotide (ODN) on the growth of three kinds of tumor cells was studied, since MTs may be involved in cell growth. When MT-I antisense ODN was added to leukemia P388 cells, cell growth was inhibited in a manner dependent on the dose and incubation time. MT-I antisense ODN was also inhibitory for other tumor cell lines, i.e. Ehrlich carcinoma and sarcoma 180. A significant decrease in the level of MT, but not of Zn, was observed in MT-I antisense ODN-treated cells. On the other hand, control ODN did not inhibit the cell growth appreciably. These results indicate that MT-I expression may be necessary for the growth and survival of these tumor cells. PMID- 9215858 TI - Running exercise may reduce risk for lung and liver cancer by inducing activity of antioxidant and phase II enzymes. AB - The effects of exercise, ethanol, and exercise plus ethanol-treatments on activity of superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase (GST) and UDP-glucuronosyl transferase (UDP-GT) in lung and liver were investigated. All treatments induced SOD and CAT activity in the lung while CAT activity was enhanced only by the combined treatments in the liver. Ethanol reduced hepatic SOD activity, while with the combined treatment SOD was normal. Exercise enhanced UDP-GT activity in liver and lung while ethanol had no effect and GST activity was induced in the liver by the combined treatment. Thus exercise may reduce risk for lung and hepatic cancer and prevent an ethanol induced increase in risk for hepatic cancer by enhancing activity of antioxidant and phase II enzymes. PMID- 9215859 TI - Sequential changes in lipoprotein lipase activity and lipaemia induced by the Yoshida AH-130 ascites hepatoma in rats. AB - The implantation of the Yoshida AH-130 ascites hepatoma to rats resulted in an exponential growth of the tumour cells followed by a late stationary phase. The tumour burden was accompanied by a dramatic decrease in body weight. Tumour growth was associated with a marked hypertriglyceridaemia during the period of exponential growth, while in the stationary phase the plasma triacylglycerol concentration was similar to that observed in the non-tumour-bearing animals. Similar increases were observed, following tumour inoculation, in the plasma concentrations of non-esterified fatty acids and glycerol, suggesting an intense lipolytic activity. These changes in lipaemia were associated with a marked decrease in LPL activity in white adipose tissue; in contrast, LPL activity was increased in the tumour-bearing animals in brown adipose tissue at day 6 following inoculation and in the heart during most of the period studied. Although the presence of the tumour did not induce any changes in blood lactate concentrations, it caused a decrease in circulating glucose; conversely, the tumour induced an important increase in the concentration of circulating ketone bodies, suggesting a metabolic adaptation of the tumour-bearing rats to glucose sparing and alternative fuel utilization. It may be suggested that the hyperlipidaemia present in the Yoshida AH-130 bearing rats is partly due to a decreased LPL activity in white adipose tissue which does not seem to be influenced by changes in insulin circulating concentrations. PMID- 9215860 TI - Deoxycytidine in human plasma: potential for protecting leukemic cells during chemotherapy. AB - Degradation of DNA produces deoxycytidine. Metabolism of deoxycytidine to dCTP inhibits phosphorylation of cytosine arabinoside (araC), fludarabine (FaraA) and 2-chlorodeoxyadenosine (CdA) by deoxycytidine kinase. This study measured plasma deoxycytidine in healthy adults and two leukemia patients and then determined how clinically relevant deoxycytidine levels would affect drug toxicity in human leukemia and lymphoma cells. Deoxycytidine was well below 0.05 microM in ten healthy persons. In the leukemia patients it was <0.05 and 0.44 microM before chemotherapy, rising to 10.3 and 5.5 microM during treatment. A broad range of clinically relevant deoxycytidine levels were high enough to profoundly decrease araC, FaraA and CdA toxicity in MOLT3, CA46 and HL60 leukemia/lymphoma cells and to change dCTP, DNA synthesis and drug incorporation into DNA in a manner consistent with prior mechanistic studies. Varying deoxycytidine levels could be an important factor influencing leukemia therapy. PMID- 9215861 TI - Lack of prognostic value of cathepsin D levels for predicting short term outcomes of breast cancer patients. AB - The value of cathepsin D determinations done on tumor cytosols in evaluating the prognosis of breast cancer patients has been debated in the literature. Our previous work suggested that cathepsin D determinations were not of prognostic value, but in that study we used immunoblotting and immunohistochemical methods rather than the more widely used double antibody immunoradiometric (IRMA) assay for measuring cathepsin levels. Here we report our results determining cathepsin D using components of a commercially available IRMA system on a large patient sample (n = 1984). Reagents from a commercially available IRMA kit were used to analyze cathepsin D levels in the cytosols of 1984 patients with breast cancer. All patients had invasive breast cancer with known tumor size and with some axillary nodes pathologically examined. Only patients with T1 and T2 tumor sizes were included. Median follow-up was 37 months. The hypothesis that high cathepsin D levels correlated with poorer outcome (poorer DFS or OS) was not confirmed, either in all patients, or in node-positive or node-negative subsets. Only in patients treated with adjuvant therapy were higher cathepsin D levels correlated with negative outcome (worsened OS, but not DFS), although given the large number of subsets analyzed this correlation may be spurious. Multivariate analyses using interaction terms did not support the concept that high cathepsin D levels correlate with resistance to adjuvant therapy. In this study evaluating the value of cathepsin D using components from a kit widely used for measuring cathepsin D levels, we conclude that cathepsin D is of doubtful value in predicting risk of early relapse or death for patients with newly diagnosed invasive breast cancer. PMID- 9215862 TI - Apoptosis bcl-2 and p53 expression and their relation to tumour stage in non small cell lung carcinomas (NSCLC). AB - Little quantitative data exist on the extent of apoptosis (genetically mediated cell deletion) and no data are available on its relation to p53 and bcl-2 expression and on its value as a prognostic factor in NSCLCs. We examined 38 NSCLCs (26 squamous, 8 adeno, 2 adenosquamous and 2 large cell carcinomas) for the frequency of apoptotic bodies by morphometric methods using haematoxylin eosin stained sections and for the bcl-2 and mutant p53 gene product expression using immunohistochemical techniques. We also evaluated the relation of apoptosis, bcl-2 and p53 expression to tumour stage and to each other. Eleven cases were in stage I, 5 were in stage II, 13 were in stage III and 9 were in stage IV. The mean apoptotic count was 9.52 (r: 2-26); 36.8% of cases were positive for bcl-2 and 76.3% of cases were positive for p53 expression. Statistical analysis did not show any correlation between tumour stage and any of the three tested parameters. There was no statistically significant relation between apoptosis and either p53 or bcl-2 expression. There are conflicting reports on the complex relationship between bcl-2, p53 and apoptosis. bcl-2 is suggested to have a prognostic value, independent from stage in SCLCs. Though we did not find any relation between stage and bcl-2 or apoptosis, it remains to be tested whether they have any independent prognostic value in larger series with survival data. PMID- 9215864 TI - Inhibitory effect of dietary curcumin on skin carcinogenesis in mice. AB - Laboratory animal model studies have suggested that curcumin may play an important role in inhibiting the process of carcinogenesis. Curcumin, the yellow pigment that is obtained from rhizomes of the plant Curcuma longa Linn (Family Zingiberaceae), is commonly used as a spice and food coloring agent. The present study was designed to investigate the chemopreventive action of dietary curcumin on 7,12-dimethylbenz[a]anthracene (DMBA)-initiated and 12-O-tetradecanoylphorbol 13-acetate (TPA)-promoted skin tumor formation in male Swiss ablino mice. At 6 weeks of age, groups of animals were fed the standard (modified AIN-76 A) diet or a diet containing 1% curcumin. At 8 weeks of age, all animals, except those in the vehicle (acetone)-treated groups, received 100 microg of DMBA dissolved in 100 microl of acetone in a single application to the skin of the back. From 1 week after DMBA application, tumor promoter (2.5 microg of TPA dissolved in 100 microl of acetone) was applied to the same areas on mouse skin twice a week for 26 weeks. All groups continued on their respective dietary regimen until the termination of the experiment. The results indicate that dietary administration of curcumin significantly inhibited the number of tumors per mouse (P < 0.05) and the tumor volume (P < 0.01). The percentage of tumor-bearing mice tended to be lower in the mice on the curcumin diet than those on the standard diet. There was no difference in growth between mice of the standard and 1% curcumin groups. The results indicate the safety and the anti-carcinogenic effect of curcumin in mice. PMID- 9215865 TI - Expression and intracellular localization of heat shock proteins in multidrug resistance of a cisplatin resistant human ovarian cancer cell line. AB - TYK-R10 is a cisplatin resistant human ovarian carcinoma cell line and showed a cross resistance to various anti-cancer drugs including adriamycin (ADR), vincristine (VCR) and etoposide, despite a lack of multidrug phenotype. Under normal conditions, various heat shock proteins (HSPs) were expressed in TYK-R10 but not in parental line (TYK-nu). Non-lethal short-term heat shock treatment induced a high tolerance for cisplatin and VCR in TYK-R10 and ADR, and VCR in TYK nu. This treatment induced and/or enhanced the expression of various types of HSPs in various intracellular localizations in both TYK-R10 and TYK-nu, with minor differences. These findings indicate that combined expression and intracellular localization of HSPs may play an important role in drug resistance of TYK-R10. PMID- 9215863 TI - Modulation of hormone-dependent transcriptional activity of the glucocorticoid receptor by the tumor suppressor p53. AB - The glucocorticoid receptor (GR) is a ligand-dependent transcription factor which regulates growth, development and metabolic functions. To test the hypothesis that the pleiotropic effect of the GR could be mediated by other transcription factors/oncogenes, the present study assessed its interaction with the tumor suppressor p53. p53 is a transcription factor which is involved in cell cycle regulation and apoptosis. We found that the wild-type p53 physically interacted with the GR and repressed the glucocorticoid-dependent transcriptional activity. In contrast, mutant p53 had no or a lesser effect depending on the type of p53 mutant. These findings raised the possibility that p53 may play an important role in modulating the activities of glucocorticoids in cells. PMID- 9215866 TI - Production of arachidonic acid metabolites by the colon adenocarcinoma cell line HT29 cl.19A and their effect on chloride secretion. AB - Eicosanoids were found in large amounts in the colonic mucosa of patients suffering from inflammatory bowel diseases and colonic adenocarcinoma. The aim of this study was to evaluate the role of the intestinal epithelial cells in the arachidonic acid metabolism and their functional response to certain eicosanoids. We used the human adenocarcinoma epithelial cell line HT29 cl.19A cell, which is an in vitro model of colon carcinoma and ion transport. These cells were found to express 5- and 15-lipoxygenase, leukotriene A4 hydrolase and cyclooxygenase-1 and -2 mRNAs. We observed an arachidonic acid metabolism via 5-lipoxygenase pathway despite the lack of FLAP mRNA expression and that certain eicosanoids such as hydroperoxy- and hydroxyeicosatetraenoic acids stimulate chloride secretion. PMID- 9215867 TI - Anti-tumor mechanisms of 3'-ethynyluridine and 3'-ethynylcytidine as RNA synthesis inhibitors: development and characterization of 3'-ethynyluridine resistant cells. AB - To discover the mechanisms of anti-tumor action of 3'-ethynyluridine (EUrd) and 3'-ethynylcytidine (ECyd), we established an EUrd-resistant variant from human fibrosarcoma HT-1080 cells. The cells were cross-resistant to ECyd. Uridine/cytidine kinase activity diminished in the resistant cells. The incorporation of EUrd and ECyd into the RNA fraction in the resistant cells was less than that of the parental cells. EUrd-triphosphate inhibited RNA synthesis by human RNA polymerase II. The results led us to conclude that EUrd and ECyd are phosphorylated by uridine/cytidine kinase to 5'-triphosphates, and that their triphosphates might inhibit RNA polymerase. PMID- 9215868 TI - Introduction of p21(Waf1/Cip1) gene into a carcinoma cell line of the uterine cervix with inactivated p53. AB - In carcinomas of the uterine cervix of which p53 is frequently inactivated by papilloma viruses, gene transfer of p21, effector of p53, is an alternative tool to suppress cell growth. We introduced the p21 gene into HeLa cells. The transfectant with p21 showed a significant growth retardation by blockage of G1 to S transfer of the cell cycle. This cell line showed significantly reduced anchorage-independent growth as well as attenuated telomerase activity. These data suggest that gene transfer of p21 is an effective tool to lead carcinoma cells with inactivated p53 into less malignant phenotype. PMID- 9215869 TI - Radiation-induced mammary tumorigenesis in pseudopregnant rats mated with vasectomized partners. AB - Pseudopregnancy was induced in Wistar-MS female rats by mating with male rats 1 month after vasectomy. Pseudopregnant rats received whole-body irradiation with 2.6 Gy gamma-rays at day 7 of pseudopregnancy and were then implanted with a DES pellet as a tumor promoter. In the control groups, virgin 2.5 month-old rats, and normal pregnant rats at day 7 of pregnancy were also exposed to the same dose of radiation. The incidence of mammary tumors in rats irradiated in pseudopregnancy (52.6%) was significantly higher than that in the irradiated virgin rats (20.8%), but was not significantly different from that observed in rats irradiated in normal pregnancy (66.7%). The appearance of the first mammary tumor in the rats irradiated in pseudopregnancy occurred 4 months earlier than that in the normal pregnancy group. The proportion of adenocarcinomas in total tumors was 16.7, 0 and 38.5% in rats irradiated in pseudopregnancy, virgin and pregnancy groups, respectively. The incidence of adenocarcinomas was not significantly different among the three groups. PMID- 9215870 TI - Reduction in NaCl-enhanced gastric carcinogenesis in rats fed a high-protein diet. AB - The effect of a purified, high protein diet on enhanced gastric carcinogenesis induced by oral administration of NaCl was investigated in Wistar rats. Rats were fed on a purified diet with an equalized caloric content, containing 8% NaCl and 25% casein (normal protein diet), or 50% casein (high protein diet) after oral treatment with N-methyl-N'-nitro-N-nitrosoguanidine for 25 weeks. In week 52, oral administration of NaCl had significantly increased the incidence and size of gastric cancer in rats fed a normal protein diet. However, NaCl had no significant effect on gastric carcinogenesis in rats fed a high protein diet. Oral administration of NaCl also caused a significant increase in tissue norepinephrine concentrations in the antral portion of the gastric wall, and increased the labeling indices of the antral epithelial cells of rats fed on a normal protein diet. However, in rats fed a high protein diet, administration of NaCl had no significant influence on these two parameters. These findings indicate that a high protein diet attenuates enhanced gastric carcinogenesis induced by the administration of NaCl, and that this effect may be related to its ability to decrease norepinephrine concentrations in the gastric wall, which subsequently decreases the proliferation of antral epithelial cells. PMID- 9215871 TI - Low frequency of c-kit expression and detection of an aberrant Kit message among Hong Kong Chinese myelogenous leukaemia patients. AB - The c-kit proto-oncogene encodes a transmembrane tyrosine kinase receptor. It is expressed by the primitive CD34 positive haemopoietic stem cells and interacts with the Kit ligand for signal transduction. It was reported to be expressed in over 80% of acute myelogenous leukaemia (AML) patients in North America and Japan. We analyzed 20 AML patients for c-kit expression using either Northern blot analysis or flow cytometry with the YB5.B8 anti-c-kit antibodies. Only 6 out of 20 AML patients expressed the c-kit mRNA or protein product. However, a previously unreported abnormal sized 1.7-1.9 kb transcript was detected in the blast cells of 1 AML patient, 1 acute mixed lineage leukaemia patient and 1 chronic myelogenous leukaemia (CML) patient in myeloblastic transformation. Our data suggested that in most Hong Kong Chinese AML patients, leukaemia transformation may have occurred at a c-kit negative stage. Alternatively, the abnormal sized c-kit transcript that was detected in some Chinese myeloid leukaemia patients may represent an aberrant c-kit receptor that plays an important role in leukaemogenesis. PMID- 9215872 TI - Direct detection of hepatitis B virus gene integrated in the Alexander cell using fluorescence in situ polymerase chain reaction. AB - Prolonged infection of hepatitis B virus (HBV) is reported to cause hepatocellular carcinoma (HCC) via liver cirrhosis. However, it is still unknown whether the HCC is induced by the HBV DNA integration or by inflammatory stimulation during the phase of liver cirrhosis. The aim of this study is to determine the intracellular or intranuclear distribution of HBV DNA with a highly sensitive assay. Here we directly detected the integration of HBV DNA by fluorescence in situ polymerase chain reaction (FISPCR). Since FISPCR products directly incorporate rhodamine-4-dUTP, the nucleus of Alexander cells integrated with HBV gene reacted with the HBV primers emits obvious fluorescence. The fluorescence values which were measured with an imaging analyzer show a significant difference between Alexander cells as compared to the controls. In conclusion, the target sequences of HBV were specifically amplified as fluorescent DNA after the present FISPCR procedure. This method could provide a novel and simple strategy for determining the quantitative role of viral DNA integration in oncogenesis. PMID- 9215873 TI - Effect of turmeric oil and turmeric oleoresin on cytogenetic damage in patients suffering from oral submucous fibrosis. AB - In vitro studies on the effect of alcoholic extracts of turmeric (TE), turmeric oil (TO) and turmeric oleoresin (TOR), on the incidence of micronuclei (Mn) in lymphocytes from normal healthy subjects showed that the test compounds did not cause any increase in the number of Mn as compared with those found in untreated controls. Further it was observed that all three compounds offered protection against benzo[a]pyrene induced increase in Mn in circulating lymphocytes. In subsequent studies, patients suffering from submucous fibrosis were given a total oral dose of TO (600 mg TO mixed with 3 g TE/day). TOR (600 mg + 3 g TE/day) and 3 g TE/day as a control for 3 months. It was observed that all three treatment modalities decreased the number of micronucleated cells both in exfoliated oral mucosal cells and in circulating lymphocytes. TOR was found to be more effective in reducing the number of Mn in oral mucosal cells (P < 0.001), but in circulating lymphocytes the decrease in Mn was comparable in all three groups. PMID- 9215874 TI - A historic moment: the discovery of the chemical transmission of embryonic inductions. PMID- 9215875 TI - Acceleration of the maturation of oligodendroblasts into oligodendrocytes and enhancement of their myelinogenic properties by a chemically defined medium. AB - Because of the importance of oligodendrocytes (OL) in forming and maintaining myelin in the CNS and the fact that the remyelination in the CNS is very limited in contrast to the peripheral nervous system, we investigated the effect of a chemically defined medium OLDEM, previously characterized by the maintenance of mature myelinating OL, on oligodendroblasts (or OL progenitors) in culture. The effect of each component of this medium as well as different combinations of them were also examined. Cultures were examined at different developmental stages immunocytochemically for developmental markers, such as transferrin, sulfatides, myelin basic protein and proteolipid protein. OLDEM accelerated the appearance of developmental markers and concomitant morphological changes. Furthermore, myelin specific enzymes such as glycerophosphorylcholine phosphodiesterase; p-nitro phenolphosphocholine phosphodiesterase; 2'3'-cyclic nucleotide 3' phosphodiesterase and UDP galactose: ceramide galactosyltransferase had enzymatic activities similar to values found in pure myelin, indicating that OLDEM allows the optimal expression of myelin-related genes. The effect of each OLDEM constituent was evaluated by immunocytochemistry and by measurement of enzymatic activities. With each single additive or multiple combinations, oligodendrocytes displayed different degrees of maturation. Deletion of selenium, glucose, and galactose severely affected cell survival as well as enzymes expression in young cultures. However, older cultures were more resistant to these deletions. Putrescine and insulin did not cause such effects on survival, but their absence affected cell maturation. None of the OLDEM additives individually supported survival and/or maturation. Enzyme assays performed on isolated myelin-like membranes or the cells soma revealed a redistribution of the activity between these fractions as the cell matured. The biological role of each of these constituents on the maturation of the oligodendroglial cells is discussed. These observations indicate that OLDEM constituents have a powerful effect on OL progenitor maturation, and membrane formation. This medium will be used for investigating the remyelination potential of adult OL progenitors. PMID- 9215876 TI - Inhibition of fatty acid beta-oxidation in rat brain cultured astrocytes exposed to the neurotoxin 3-nitropropionic acid. AB - We describe the effects of the neurotoxin 3-nitropropionic acid (3-NPA) on fatty acid oxidation in neonatal rat brain astrocytes in primary culture, using a sensitive assay for beta-oxidation which depends on the release of 3H2O from [9,10(n)-3H]palmitic acid. 3-NPA is a suicide inhibitor of succinate dehydrogenase, a constituent of both Krebs cycle and complex II of the mitochondrial respiratory chain. It is widely distributed in plants and fungi. Neurotoxicity of 3-NPA to humans and animals, leading to selective neuronal cell death, appears mediated by the reduced level of ATP induced by the toxin. We demonstrated that 3-NPA can also impair energy metabolism in astrocytes. Exposure of astroglial cells in culture to 3-NPA leads to inhibition of the release of 3H2O from [9,10(n)-3H]palmitic acid. Addition of 2 mM 3-NPA to the culture medium caused a rapid decrease in beta-oxidation activity, which reached a plateau after 90 min. This inhibition was concentration-dependent. Concentration as low as 0.05 mM for 5 h significantly decreased beta-oxidation activity (25% inhibition). Half maximal inhibition was obtained after treatment with 0.5 mM 3-NPA, and 3 mM induced a maximal response (63% inhibition) 3-NPA is clearly a potent inhibitor of beta-oxidation activity. We also show that 3-NPA 3 mM inhibits partially complex II (succinate ubiquinone reductase) and aspartate aminotransferase by 60 and 49% after 4 h treatment respectively. It has been shown that fatty acid is the preferred substrate for energy production in cultured astrocytes from developing brain. As astrocytes may also provide substrates alternative for energy metabolism in neurons and oligodendrocytes, it is likely that the inhibition of beta-oxidation by 3-NPA may contribute significantly to the damage induced by this toxin in the central nervous system. PMID- 9215877 TI - Tissue culture model of Krabbe's disease: psychosine cytotoxicity in rat oligodendrocyte culture. AB - Krabbe's disease (globoid cell leukodystrophy) is a progressive cerebral degenerative disease of infancy characterized by severe myelin loss and the presence of globoid bodies in the white matter. Previous studies have suggested that psychosine is the causative agent for the pathogenesis of Krabbe's disease. In the present study, we investigated psychosine-induced injury and cell death of oligodendrocytes in enriched cultures of oligodendrocytes prepared from 3-week old rat brain. The psychosine concentration sufficient to induce 50% cell death in oligodendrocytes was 30 micrograms/ml in the medium containing serum and 10 micrograms/ml in the serum-free medium. When oligodendrocytes were exposed to psychosine in the presence of phorbol esters, insulin, insulin-like growth factor I (IGF-I), demethylsulfoxide, or serum albumin, the survival of oligodendrocytes was greatly increased. These results indicate that psychosine cytotoxicity against oligodendrocytes is blocked by phorbol esters, insulin, and IGF-I through activation of protein kinase-C, by dimethylsulfoxide through activation of beta galactosidase, and by albumin through its binding to psychosine. PMID- 9215878 TI - Development of galanin- and enkephalin-like immunoreactivities in the sympathoadrenal lineage of the avian embryo. In vivo and in vitro studies. AB - The neurochemical differentiation of the sympathoadrenal nervous system has been analyzed by focusing on the developmental expression of two neuropeptides, galanin and enkephalin. Both peptides are expressed early in the formation of the sympathetic ganglia and adrenal gland. Expression in the adrenal persists during embryogenesis to hatching while expression in the sympathetic is lost as sympathoblasts differentiate into neurons. Galanin expression and its modulation by nerve growth factor (NGF) and dexamethasone (Dex) was also studied in vitro. Differential effects of these factors were found on adrenal versus sympathetic cultures. However, the results coincided with proposed role of the factors in inducing either neuronal properties (NGF) or chromaffin characteristics (Dex). PMID- 9215879 TI - Jimpy-4J mouse has a missense mutation in exon 2 of the Plp gene. AB - We previously showed that the jimpy-4J mouse mutation is located on the X chromosome, in or closely linked to the proteolipid protein (Plp) gene. The phenotype is characterized by the most severe hypomyelination of any of the naturally occurring myelin mutant mice, sharp reduction in oligodendrocyte number, and virtual absence of PLP protein. Affected animals show tremor, seizures, and die at about 24 postnatal days. We now report that sequencing of Plp genomic and cDNAs identifies a single nucleotide substitution in exon 2 that predicts an Ala38Ser substitutions in a hydrophilic region of PLP/DM20 protein close to a transmembrane domain. This mutation occurs in a very different region of the mouse Plp gene than that jimpy-msd mutations, yet all three produce qualitatively similar phenotypes. PMID- 9215880 TI - The neonatal neurotoxicity of monosodium L-glutamate on the sexually dimorphic nucleus of the preoptic area in rats. AB - The neurotoxic effect of monosodium L-glutamate (MSG) on the morphologies in the darkly stained sexually dimorphic nucleus of the preoptic area (SDN-POA) and the lighter-staining surrounding area (non-SDN-POA) within the medial preoptic nucleus (MPN) was evaluated. Male and female Long-Evans rats were used. MSG (4 mg/g of body weight) was administered subcutaneously to pups on days 1 and 3 postnatally. Normal saline was used as the vehicle. At the age of 6 months, the rats were sacrificed and the brain tissues were fixed for histological examination. The morphological changes, i.e., total volume, density, total neuron number, neuronal nuclear volume (NNV) and ratio of pyknosis, of the SDN-POA and non-SDN-POA within the MPN, were estimated using the AMS VIDS III semiautomatic image-analytic system. The results indicate that neonatal MSG treatment caused significant neuronal loss and decreases in total volume of the SDN-POA and non SDN-POA of male and female rats. However, only the SDN-POA of MSG-treated male rats showed a significant increase of pyknosis and decrease of neuronal density. A significant enlargement of NNV in the SDN-POA and non-SDN-POA was observed in the MSG-treated male rats. These results indicate that the MPN shows sex-specific and area-specific changes after neonatal neurotoxicity due to MSG. PMID- 9215881 TI - A monoclonal antibody recognizing nestin and the phosphorylated form of neurofilament-M. AB - In the course of an immunohistochemical screening of antibodies raised against a postnatal day 7 rat brain membrane fraction, we obtained a monoclonal antibody that recognizes both neurofilament-M, a type VI intermediate filament protein, and nestin, a type IV intermediate filament protein. In newborn rat cerebellum, the 5A7 antibody stained Bergmann glial fibers and subsets of axons simultaneously, but in adult cerebellum it only stained neuronal elements. Upon immunoblot analysis of neonatal rat cerebellum, 5A7 gave two prominent bands corresponding to 140 and > 200 kDa. The former was identified as neurofilament-M and the latter as nestin. Dephosphorylation of neurofilament-M resulted in a dramatic reduction in the immunoreactivity, indicating that 5A7 recognized the phosphorylated form of neurofilament-M. Immunoblot analysis of V8 protease digests of neurofilament-M revealed that the epitope on neurofilament-M is located on the carboxy-half of the tail domain of neurofilament-M. These results indicate that 5A7 is the first monoclonal antibody which recognizes a common or similar epitope on different classes of intermediate filament proteins other than the conserved rod domain. The presence of a common or similar epitope on both nestin and neurofilament-M recognized by 5A7 may suggest the possibility that nestin and neurofilament-M have a common functional motif. PMID- 9215882 TI - Extracellular-matrix molecules expressed by innervatable muscle. AB - Embryonic and adult muscle express distinct complements of proteins. Transplant experiments have shown that embryonic and denervated muscles can form synapses with foreign neurites, while normal innervated mature muscles cannot, except at the synaptic area. To identify molecular differences between 'innervatable' embryonic muscle and 'non-innervatable' mature muscle, we used a differential immunization method to make monoclonal antibodies that recognize antigens present in embryonic muscle rather than normal mature muscle. Four independent monoclonal antibodies that stain embryonic and mature muscle sections differentially have been obtained. One stains the entire embryonic muscle cell surface but only the synaptic area in mature muscle; 3 stain the entire embryonic muscle cell surface but do not stain mature muscle. The antibodies also stain other embryonic tissues at several stages. The antigens are concentrated in basal laminae and in an extracellular-matrix (ECM) fraction, indicating that they are ECM molecules. The temporal expression of all 4 antigens in developing muscle is coincident with muscle innervation. All antigens exhibit temporal and tissue distributions different from those of any reported muscle proteins, suggesting that novel antigens underlie these patterns. These results confirm that the immature muscle ECM has a distinct molecular composition, and suggest that the presence of these antigens may be part of the molecular basis for the innervatability of embryonic muscle. PMID- 9215883 TI - Effects of short- and long-term withdrawal from gestational cocaine treatment on maternal behavior and aggression in Sprague-Dawley rats. AB - Pregnant rats were treated with 30 mg/kg per day cocaine or normal saline either throughout gestation (GD 1-20, cocaine and saline withdrawal) or throughout the gestation and continuing into lactation for 10 days postpartum (cocaine and saline nonwithdrawal). All cocaine-treated dams exhibited more disruptions in the onset of maternal behavior (retrieval, licking, crouching) and were more aggressive (threats and attacks) towards an intruder on postpartum day 6 than saline-treated dams. There were no significant differences in these behaviors between withdrawn and nonwithdrawn cocaine-treated dams. These findings indicate that changes in maternal behavior following chronic moderate cocaine treatment are not simply the result of withdrawal from cocaine treatment following gestation and that other possible mechanisms should be examined. PMID- 9215884 TI - Recombination and population structure in Escherichia coli. PMID- 9215885 TI - Pathways for homologous recombination between chromosomal direct repeats in Salmonella typhimurium. AB - Homologous recombination pathways probably evolved primarily to accomplish chromosomal repair and the formation of and resolution of duplications by sister chromosome exchanges. Various DNA lesions initiate these events. Classical recombination assays, involving bacterial sex, focus attention on double-strand ends of DNA. Sexual exchanges, initiated at these ends, depend on the RecBCD pathway. In the absence of RecBCD function, mutation of the sbcB and sbcC genes activates the apparently cryptic RecF pathway. To provide a more general view of recombination, we describe an assay in which endogenous DNA damage initiates recombination between chromosomal direct repeats. The repeats flank markers conferring lactose utilization (Lac+) and ampicillin resistance (ApR); recombination generates Lac-ApS segregants. In this assay, the RecF pathway is not cryptic; it plays a major role without sbcBC mutations. Others have proposed that single-strand gaps are the natural substrate for RecF-dependent recombination. Supporting this view, recombination stimulated by a double-strand break (DSB) in a chromosomal repeat depended on RecB function, not RecF function. Without RecBCD function, sbcBC mutations modified the RecF pathway and allowed it to catalyze DSB-stimulated recombination. Sexual recombination assays overestimate the importance of RecBCD and DSBs, and underestimate the importance of the RecF pathway. PMID- 9215886 TI - Microsatellite instability in yeast: dependence on the length of the microsatellite. AB - One of the most common microsatellites in eukaryotes consists of tandem arrays [usually 15-50 base pairs (bp) in length] of the dinucleotide GT. We examined the rates of instability for poly GT tracts of 15, 33, 51, 99 and 105 bp in wild-type and mismatch repair-deficient strains of Saccharomyces cerevisiae. Rates of instability increased more than two orders of magnitude as tracts increased in size from 15 to 99 bp in both wild-type and msh2 strains. The types of alterations observed in long and short tracts in wild-type strains were different in two ways. First, tracts > or = 51 bp had significantly more large deletions than tracts < or = 33 bp. Second, for the 99- and 105-bp tracts, almost all events involving single repeats were additions; for the smaller tracts, both additions and deletions of single repeats were common. PMID- 9215887 TI - Isolation of COM1, a new gene required to complete meiotic double-strand break induced recombination in Saccharomyces cerevisiae. AB - We have designed a screen to isolate mutants defective during a specific part of meiotic prophase I of the yeast Saccharomyces cerevisiae. Genes required for the repair of meiotic double-strand breaks or for the separation of recombined chromosomes are targets of this mutant hunt. The specificity is achieved by selecting for mutants that produce viable spores when recombination and reductional segregation are prevented by mutations in SPO11 and SPO13 genes, but fail to yield viable spores during a normal Rec+ meiosis. We have identified and characterized a mutation com1-1, which blocks processing of meiotic double-strand breaks and which interferes with synaptonemal complex formation, homologous pairing and, as a consequence, spore viability after induction of meiotic recombination. The COM1/SAE2 gene was cloned by complementation, and the deletion mutant has a phenotype similar to com1-1, com1/sae2 mutants closely resemble the phenotype of rad50S, as assayed by phase-contrast microscopy for spore formation, physical and genetic analysis of recombination, fluorescence in situ hybridization to quantify homologous pairing and immunofluorescence and electron microscopy to determine the capability to synapse axial elements. PMID- 9215888 TI - A general method for identifying recessive diploid-specific mutations in Saccharomyces cerevisiae, its application to the isolation of mutants blocked at intermediate stages of meiotic prophase and characterization of a new gene SAE2. AB - We describe a general new approach for identifying recessive mutations that affect diploid strains of yeast Saccharomyces cerevisiae and the application of this method to the identification of mutations that confer an intermediate block in meiotic prophase chromosome metabolism. The method uses a temperature sensitive conjugation mutation ste7-1 in combination with homothallism. The mutations of interest confer a defect in spore formation that is dependent upon a gene required for initiation of meiotic recombination and development of meiosis specific chromosome structure (SPO11). Identified in this screen were null mutations of the DMC1 gene, nonnull mutations of RAD50 (rad50S), and mutations in three new genes designed SAE1, SAE2 and SAE3 (Sporulation in the Absence of Spo Eleven). Molecular characterization of the SAE2 gene and characterization of meiotic and mitotic phenotypes of sae2 mutants are also presented. The phenotypes conferred by a sae2 null mutation are virtually indistinguishable from those conferred by the previously identified nonnull mutations of RAD50 (rad50S). Most notably, both mutations confer only weak sensitivity to the radiomimetic agent methyl methane sulfonate (MMS) but completely block resection and turnover of meiosis-specific double-strand breaks. These observations provide further evidence that this constellation of phenotypes identifies a specific molecular function. PMID- 9215891 TI - Saccharomyces cerevisiae PAC2 functions with CIN1, 2 and 4 in a pathway leading to normal microtubule stability. AB - The products of the Saccharomyces cerevisiae CIN1, CIN2 and CIN4 genes participate in a nonessential pathway required for normal microtubule function. In this article, we demonstrate that the product of PAC2 also functions in this pathway. PAC2 deletion mutants displayed phenotypes and genetic interactions similar to those caused by cin1 delta, cin2 delta and cin4 delta. These include cold-sensitive microtubule structures and sensitivity to the microtubule depolymerizing agent benomyl. Involvement in a common functional pathway is indicated by the observation that all double mutant recombinations are viable and no more affected than any single mutant. In addition, extra copies of CIN1 were found to suppress the benomyl sensitivity of pac2 delta, cin2 delta and cin4 delta, but not that caused by other mutations that affect microtubule function. Cin1p and Pac2p were found to be related in sequence to mammalian proteins that aid in the folding of beta-tubulin into an assembly-competent state. Alleles of CIN1 were identified that could suppress the benomyl sensitivity of cin4-4 in a highly specific fashion. Our findings suggest that the guanine nucleotide-binding Cin4p interacts with Cin1p and regulates its tubulin folding activity. PMID- 9215889 TI - Mutations in Saccharomyces cerevisiae that block meiotic prophase chromosome metabolism and confer cell cycle arrest at pachytene identify two new meiosis specific genes SAE1 and SAE3. AB - Two new meiosis-specific genes, SAE1 and SAE3, have been identified in a screen for mutations that confer an intermediate block in meiotic prophase. Such mutations confer a block to spore formation that is circumvented by addition of a mutation that eliminates meiotic recombination initiation and other aspects of chromosome metabolism, i.e., spo11. We show that sae1-1 and sae3-1 mutations each confer a distinct defect in meiotic recombination. sae1-1 produces recombinants but very slowly and ultimately to less than half the wild-type level; sae3-1 makes persistent hyper-resected meiotic double-strand breaks and has a severe defect in formation of recombinants. Both mutants arrest at the pachytene stage of meiotic prophase, sae1-1 temporarily and sae3-1 permanently. The phenotypes conferred by sae3-1 are similar to those conferred by mutation of the yeast RecA homologue DMC1, suggesting that SAE3 and DMC1 act at the same step(s) of chromosome metabolism. These results provide further evidence that intermediate blocks to prophase chromosome metabolism cause cell-cycle arrest. SAE1 encodes a 208-residue protein homologous to vertebrate mRNA cap-binding protein 20. SAE3 corresponds to a meiosis-specific RNA encoding an unusually short open reading frame of 50 codons. PMID- 9215890 TI - A 140-bp-long palindromic sequence induces double-strand breaks during meiosis in the yeast Saccharomyces cerevisiae. AB - Palindromic sequences have the potential to form hairpin or cruciform structures, which are putative substrates for several nucleases and mismatch repair enzymes. A genetic method was developed to detect such structures in vivo in the yeast Saccharomyces cerevisiae. Using this method we previously showed that short hairpin structures are poorly repaired by the mismatch repair system in S. cerevisiae. We show here that mismatches, when present in the stem of the hairpin structure, are not processed by the repair machinery, suggesting that they are treated differently than those in the interstrand base-paired duplex DNA. A 140 bp-long palindromic sequence, on the contrary, acts as a meiotic recombination hotspot by generating a site for a double-strand break, an initiator of meiotic recombination. We suggest that long palindromic sequences undergo cruciform extrusion more readily than short ones. This cruciform structure then acts as a substrate for structure-specific nucleases resulting in the formation of a double strand break during meiosis in yeast. In addition, we show that residual repair of the short hairpin structure occurs in an MSH2-independent pathway. PMID- 9215892 TI - Mating type in Chlamydomonas is specified by mid, the minus-dominance gene. AB - Diploid cells of Chlamydomonas reinhardtii that are heterozygous at the mating type locus (mt+/mt-) differentiate as minus gametes, a phenomenon known as minus dominance. We report the cloning and characterization of a gene that is necessary and sufficient to exert this minus dominance over the plus differentiation program. The gene, called mid, is located in the rearranged (R) domain of the mt- locus, and has duplicated and transposed to an autosome in a laboratory strain. The imp11 mt- mutant, which differentiates as a fusion-incompetent plus gamete, carries a point mutation in mid. Like the fus1 gene in the mt+ locus, mid displays low codon bias compared with other nuclear genes. The mid sequence carries a putative leucine zipper motif, suggesting that it functions as a transcription factor to switch on the minus program and switch off the plus program of gametic differentiation. This is the first sex-determination gene to be characterized in a green organism. PMID- 9215893 TI - Phenotypic and genetic analysis of "Chameleon," a paramecium mutant with an enhanced sensitivity to magnesium. AB - Three mutant strains of Paramecium tetraurelia with an enhanced sensitivity to magnesium have been isolated. These new "Chameleon" mutants result from partial- or codominant mutations at a single locus, Cha. Whereas the wild type responded to 5 mM Mg2+ by swimming backward for 10-15 sec, Cha mutants responded with approximately 30 sec backward swimming. Electrophysiological analysis suggested that this behavior may be caused by slowing in the rate at which a Mg(2+) specific ion conductance deactivates following membrane excitation. This would be consistent with an observed increase in the sensitivity of Cha mutants to nickel poisoning, since Ni2+ is also able to enter the cell via this pathway. More extensive behavioral analysis showed that Cha cells also overresponded to Na+, but there was no evidence for a defect in intracellular Ca2+ homeostasis that might account for a simultaneous enhancement of both the Mg2+ and Na+ conductances. The possibility that the Cha locus may encode a specific regulator of the Mg(2+)- and Na(+)-permeabilities is considered. PMID- 9215894 TI - Genetic and environmental responses to temperature of Drosophila melanogaster from a latitudinal cline. AB - Field-collected Drosophila melanogaster from 19 populations in Eastern Australia were measured for body size traits, and the measurements were compared with similar ones on flies from the same populations reared under standard laboratory conditions. Wild caught flies were smaller, and latitudinal trends in size were greater. Reduced size was caused by fewer cells in the wing, and the steeper cline by greater variation in cell area. The reduction in size in field-collected flies may therefore have been caused by reduced nutrition, and the steeper cline may have been caused by an environmental response to latitudinal variation in temperature. No evidence was found for evolution of size traits in response to laboratory culture. The magnitude of phenotypic plasticity in response to temperature of development time, body size, cell size and cell number was examined for six of the populations, to test for latitudinal variation in plasticity. All characters were plastic in response to temperature. Total development time showed no significant latitudinal variation in plasticity, although larval development time showed a marginally significant effect, with most latitudinal variation at intermediate rearing temperatures. Neither thorax length nor wing size and its cellular components showed significant latitudinal variation in plasticity. PMID- 9215895 TI - Polymorphism for Y-linked suppressors of sex-ratio in two natural populations of Drosophila mediopunctata. AB - In several Drosophila species there is a trait known as "sex-ratio": males carrying certain X chromosomes (called "SR") produce female biased progenies due to X-Y meiotic drive. In Drosophila mediopunctata this trait has a variable expression due to Y-linked suppressors of sex-ratio expression, among other factors. There are tow types of Y chromosomes (suppressor and nonsuppressor) and two types of SR chromosomes (suppressible and unsuppressible). Sex-ratio expression is suppressed in males with the SRsuppressible/Ysuppressor genotype, whereas the remaining three genotypes produce female biased progenies. Now we have found that approximately 10-20% of the Y chromosomes from two natural populations 1500 km apart are suppressors of sex-ratio expression. Preliminary estimates indicate that Ysuppressor has a meiotic drive advantage of 6% over Ynonsuppressor. This Y polymorphism for a nonneutral trait is unexpected under current population genetics theory. We propose that this polymorphism is stabilized by an equilibrium between meiotic drive and natural selection, resulting from interactions in the population dynamics of X and Y alleles. Numerical simulations showed that this mechanism may stabilize nonneutral Y polymorphisms such as we have found in D. mediopunctata. PMID- 9215896 TI - A sex-influenced modifier in Drosophila that affects a broad spectrum of target loci including the histone repeats. AB - A second chromosomal trans-acting modifier, Lightener of white (Low), modulates the phenotypic expression of various alleles of the white eye color gene. This modifier has an unusually broad spectrum of affected genes including white, brown, scarlet and the eye developmental genes, Bar and Lobe. In addition, Low weakly suppresses position effect variegation. Northern blot hybridization with different X and autosomal probes reveals that Low modulates genes of independent expression patterns. Interestingly, many of the modulations of gene expression are developmentally restricted and differ in intensity between the sexes. Low also elevates the expression of the histone tandem repeats in three distinct developmental stages. A deficiency encompassing the histone cluster reduces their transcript levels and significantly alters the expression of some of the tested genes. Thus, Low is a modifier that plays a role in modulating the expression of genes governing various processes including pigment deposition, eye development, chromosomal proteins and position effect variegation. PMID- 9215897 TI - Topological constraints on transvection between white genes within the transposing element TE35B in Drosophila melanogaster. AB - The transposable element TE35B carries two copies of the white (w) gene at 35B1.2 on the second chromosome. These w genes are suppressed in zeste-1 (z1) mutant background in a synapsis-dependent manner. Single-copy derivatives of the original TE35B stock give red eyes when heterozygous, but zeste eyes when homozygous. TE35B derivatives carrying single, double or triple copies of w were crossed to generate flies carrying from two to five ectopic w genes. Within this range, z1-mediated suppression is insensitive to copynumber and does not distinguish between w genes that are in cis or in trans. Suppression does not require the juxtaposition of even numbers of w genes, but is extremely sensitive to chromosomal topology. When arranged in a tight cluster, in triple-copy TE derivatives, w genes are nonsuppressible. Breakpoints falling within TE35B and separating two functional w genes act as partial suppressors of z1. Similarly, breakpoints immediately proximal or distal to both w genes give partial suppression. This transvection-dependent downregulation of w genes may result from mis-activation of the X-chromosome dosage compensation mechanism. PMID- 9215898 TI - Point mutations within and outside the homeodomain identify sequences required for proboscipedia homeotic function in Drosophila. AB - The Drosophila homeotic gene proboscipedia (pb) encodes a homeodomain protein homologous to vertebrate HoxA2/B2 required for adult mouthparts formation. A transgenic Hsp70-pb (HSPB) element that rescues pb mutations also induces the dominant transformation of antennae to maxillary palps. To identify sequences essential to PB protein function, we screened for EMS-induced HSPB mutations leading to phenotypic reversion of the HSPB transformation. Ten revertants harbor identified point mutations in HSPB coding sequences. The point mutations that remove all detectable phenotypes in vivo reside either within the homeodomain or, more unexpectedly, in evolutionarily nonconserved regions outside the homeodomain. Two independent homeodomain mutations that change the highly conserved Arginine-5 in the N-terminal hinge show effects on adult eye development, suggesting a previously unsuspected role for Arg5 in functional specificity. Three additional revertant mutations outside the homeodomain reduce but do not abolish PB+ activity, identifying protein elements that contribute quantitatively to pb function. This in vivo analysis shows that apart from the conserved motifs of PB, other elements throughout the protein make important contributions to homeotic function. PMID- 9215899 TI - A new enhancer of position-effect variegation in Drosophila melanogaster encodes a putative RNA helicase that binds chromosomes and is regulated by the cell cycle. AB - In Drosophila melanogaster, position-effect variegation of the white gene has been a useful phenomenon by which to study chromosome structure and the genes that modify it. We have identified a new enhancer of variegation locus, Dmrnahel (hel). Deletion of mutation of hel enhances white variegation, and this can be reversed by a transformed copy of hel+. In the presence of two endogenous copies, the transformed hel+ behaves as a suppressor of variegation. hel is an essential gene and functions both maternally and zygotically. The HEL protein is similar to known RNA helicases, but contains an unusual variant (DECD) of the DEAD motif common to these proteins. Potential HEL homologues have been found in mammals, yeast and worms. HEL protein associates with salivary gland chromosomes and locates to nuclei of embryos and ovaries, but disappears in mitotic domains of embryos as chromosomes condense. We propose that the HEL protein promotes an open chromatin structure that favors transcription during development by regulating the spread of heterochromatin, and that HEL is regulated by, and may have a role in, the mitotic cell cycle during embryogenesis. PMID- 9215901 TI - Population dynamics inferred from temporal variation at microsatellite loci in the selfing snail Bulinus truncatus. AB - We analyzed short-term forces acting on the genetics of subdivided populations based on a temporal survey of the microsatellite variability in the hermaphrodite freshwater snail Bulinus truncatus. This species inhabits temporary habitats, has a short generation time and exhibits variable rates of selfing. We studied the variability over three sampling dates in 12 Sahelian populations (1161 individuals). Classical genetic parameters (estimators of Ho, He, f, selfing rate and Fst) showed limited change over time whereas important temporal changes of allelic frequencies were detected for 10 of the ponds studied. These variations are not easily explained by selection, sampling drift and genetic drift alone and may be due to periodic migration. Indeed the habitats occupied by the populations studied are subject to large temporal fluctuations owing to annual cycles of drought and flood. In such ponds our results support a demographic model of population expansions and contractions under which available habitats, after the rainy season, are colonized by individuals originating from a smaller number of refuges (areas that never dry out in the deepest parts of the ponds). In contrast, selfing appeared to be an important force affecting the genetic structure in permanent ponds. PMID- 9215900 TI - Mapping a quantitative trait locus involved in melanotic encapsulation of foreign bodies in the malaria vector, Anopheles gambiae. AB - A Plasmodium-refractory strain of Anopheles gambiae melanotically encapsulates many species of Plasmodium, whereas wild-type mosquitoes are usually susceptible. This encapsulation trait can also be observed by studying the response of refractory and susceptible strains to intrathoracically injected CM-Sephadex beads. We report the results of broad-scale quantitative trait locus (QTL) mapping of the encapsulation trait using the bead model system. Interval mapping using the method of maximum likelihood identified one major QTL, Pen1. The 13.7 cM interval containing Pen1 was defined by marker AGH157 at 8E and AGH46 at 7A on 2R. Pen1 was associated with a maximum LOD score of 9.0 and accounted for 44% of the phenotypic variance in the distribution of phenotypes in the backcross. To test if this QTL is important for encapsulation of Plasmodium berghei, F2 progeny were infected with P. berghei and evaluated for degree of parasite encapsulation. For each of the two markers that define the interval containing Pen1, a significant difference of encapsulation was seen in progeny with at least one refractory allele in contrast with homozygous susceptible progeny. These results suggest that Pen1 is important for melanotic encapsulation of Plasmodium as well as beads. PMID- 9215902 TI - Hybridogenetic reproduction and maternal ancestry of polyploid Iberian fish: the Tropidophoxinellus alburnoides complex. AB - Iberian minnows collectively known as the Tropidophoxinellus alburnoides Steindachner complex comprise diploid and polyploid forms with highly female biased sex ratios. Previous investigators suggested that all-female clonal reproduction and interspecific hybridization may occur in this complex. We examined nuclear (allozymes) and cytoplasmic genes (mtDNA) to assess the evolutionary origins, relationships, and reproductive modes of T. alburnoides from western Spain. The multi-locus allozyme data clearly revealed the hybrid nature of all polyploid forms of this fish and some diploid forms as well. Diagnostic markers identified fish from the genus Leuciscus as the paternal ancestor of hybrids in the Duero and Guadiana River Basins. Additionally, analysis of nuclear markers revealed that hybridogenetic reproduction occurs in the diploid and triploid hybrids. The hybrids fully express the paternal Leuciscus genome and then discard it during oogenesis. Hybridogenetic ova contain only maternal nuclear genes and mtDNA from a non-hybrid T. alburnoides ancestor. Apparently diploid and triploid hybrids of T. alburnoides persist as sperm parasites on males of a sexually reproducing Leuciscus host species. PMID- 9215904 TI - Estimating meiotic exchange patterns from recombination data: an application to humans. AB - We present analytical methods to estimate the recombinational history of chromosomes in a human population. Our analysis, similar to those utilized in Drosophila, can be used to construct meiotic maps based upon crossover frequencies observed in family data. We apply this method of exchange estimation to a population of paternally and maternally inherited chromosomes 21. The patterns of chromosomal exchange estimated by this type of analysis are comparable to those obtained by the more technically difficult method of cytologically counting chiasmata among human male meiotic events (sperm). This type of analysis can be applied to both male and female meiosis, circumventing many technical problems inherent to cytological counting. Moreover, the distribution of exchange locations along a chromosome for each exchange type (i.e., single, double, or triple exchanges) can be examined individually, an advantage compared to examination of genetic maps that only provide a summary of these distributions. We discuss how this analysis can be used to examine various assumptions concerning meiotic exchange in humans and investigate properties of the analysis that contribute to the accuracy of the results. PMID- 9215903 TI - The complete DNA sequence of the mitochondrial genome of a "living fossil," the coelacanth (Latimeria chalumnae). AB - The complete nucleotide sequence of the 16,407-bp mitochondrial genome of the coelacanth (Latimeria chalumnae) was determined. The coelacanth mitochondrial genome order is identical to the consensus vertebrate gene order which is also found in all ray-finned fishes, the lungfish, and most tetrapods. Base composition and codon usage also conform to typical vertebrate patterns. The entire mitochondrial genome was PCR-amplified with 24 sets of primers that are expected to amplify homologous regions in other related vertebrate species. Analyses of the control region of the coelacanth mitochondrial genome revealed the existence of four 22-bp tandem repeats close to its 3' end. The phylogenetic analyses of a large data set combining genes coding for rRNAs, tRNAs, and proteins (16,140 characters) confirmed the phylogenetic position of the coelacanth as a lobe-finned fish; it is more closely related to tetrapods than to ray-finned fishes. However, different phylogenetic methods applied to this largest available molecular data set were unable to resolve unambiguously the relationship of the coelacanth to the two other groups of extant lobe-finned fishes, the lungfishes and the tetrapods. Maximum parsimony favored a lungfish/coelacanth or a lungfish/tetrapod sistergroup relationship depending on which transversion:transition weighting is assumed. Neighbor-joining and maximum likelihood supported a lungfish/tetrapod sistergroup relationship. PMID- 9215906 TI - Evolution of repeated sequence arrays in the D-loop region of bat mitochondrial DNA. AB - Analysis of mitochondrial DNA control region sequences from 41 species of bats representing 11 families revealed that repeated sequence arrays near the tRNA-Pro gene are present in all vespertilionine bats. Across 18 species tandem repeats varied in size from 78 to 85 bp and contained two to nine repeats. Heteroplasmy ranged from 15% to 63%. Fewer repeats among heteroplasmic than homoplasmic individuals in a species with up to nine repeats indicates selection may act against long arrays. A lower limit of two repeats and more repeats among heteroplasmic than homoplasmic individuals in two species with few repeats suggests length mutations are biased. Significant regressions of heteroplasmy, theta and pi, on repeat number further suggest that repeat duplication rate increases with repeat number. Comparison of vespertilionine bat consensus repeats to mammal control region sequences revealed that tandem repeats of similar size, sequence and number also occur in shrews, cats and bighorn sheep. The presence of two conserved protein-binding sequences in all repeat units indicates that convergent evolution has occurred by duplication of functional units. We speculate that D-loop region tandem repeats may provide signal redundancy and a primitive repair mechanism in the event of somatic mutations to these binding sites. PMID- 9215905 TI - Discordant phylogeographic patterns between the Y chromosome and mitochondrial DNA in the house mouse: selection on the Y chromosome? AB - We have compared patterns of geographic variation and molecular divergence of mitochondrial DNA (mtDNA) and Y chromosome over the range of the different subspecies of Mus musculus. MtDNA was typed for 305 nucleotides in the control region, the Y chromosome for 834 base pairs (bp) in Zfy introns and 242 bp in Sry, a Zfy2 18-bp deletion, and two microsatellites. Apparent discrepancies exist between the distributions of the lineages of mtDNA and of the two major Y chromosome lineages thus defined: some subspecies share the same mtDNA lineage but have different Y-chromosome lineages or vice versa. One microsatellite reveals a geographically clustered variation inside the distribution of each Y chromosome lineage, showing that new Y-chromosome variants can rapidly spread locally. The two major Y-chromosome lineages have a divergence time only about one fourth of that between mtDNA lineages. Although this recent coalescence of the Y chromosomes between subspecies could partly be due to a lower ancestral polymorphism of the Y chromosome, it suggests that secondary introgression after the radiation of the subspecies might have occurred. There is evidence that the differentiation of the Y-chromosome lineages contributes to partial reproductive isolation between subspecies, and patterns of molecular evolution suggest that selection has played a role in the rapid spread across subspecies. PMID- 9215907 TI - The mouse Clock mutation behaves as an antimorph and maps within the W19H deletion, distal of Kit. AB - Clock is a semidominant mutation identified from an N-ethyl-N-nitrosourea mutagenesis screen in mice. Mice carrying the Clock mutation exhibit abnormalities of circadian behavior, including lengthening of endogenous period and loss of rhythmicity. To identify the gene affected by this mutation, we have generated a high-resolution genetic map (> 1800 meioses) of the Clock locus. We report that Clock is 0.7 cM distal of Kit on mouse chromosome 5. Mapping shows that Clock lies within the W19H deletion. Complementation analysis of different Clock and W19H compound genotypes indicates that the Clock mutation behaves as an antimorph. This antimorphic behavior of Clock strongly argues that Clock defines a gene centrally involved in the mammalian circadian system. PMID- 9215908 TI - Genotype selection to rapidly breed congenic strains. AB - Congenic strains can now be constructed guided by the transmission of DNA markers. This allows not only selection for transmission of a desired, donor derived differential region but also selection against the transmission of unwanted donor origin genomic material. The additional selection capacity should allow congenic strains to be produced in fewer generations than is possible with random backcrosses. Here, we consider modifications of a standard backcross breeding scheme to produce congenic mice by the inclusion of genotype-based selective breeding strategies. Simulation is used to evaluate the consequences of each strategy on the number of chromosomes that contain unwanted, donor-derived genetic material and the average length of this unwanted donor DNA for each backcross generation. Our prototypic strategy was to choose a single mouse to sire each generation using criteria designed to select against the transmission of chromosomes, other than the one containing the replacement genomic region, that contain any donor origin sequence at all. This chromosome elimination strategy resulted in an average of 16.4 chromosomes free of donor DNA in mice of the third backcross (N3) generation. A strategy based solely on positive selection for the replacement region required six backcross generations to achieve the same results. PMID- 9215909 TI - Microsatellite DNA variation and the evolution, domestication and phylogeography of taurine and zebu cattle (Bos taurus and Bos indicus). AB - Genetic variation at 20 microsatellite loci was surveyed to determine the evolutionary relationships and molecular biogeography of 20 different cattle populations from Africa, Europe and Asia. Phylogenetic reconstruction and multivariate analysis highlighted a marked distinction between humpless (taurine) and humped (zebu) cattle, providing strong support for a separate origin for domesticated zebu cattle. A molecular clock calculation using bison (Bison sp.) as an outgroup gave an estimated divergence time between the two subspecies of 610,000-850,000 years. Substantial differences in the distribution of alleles at 10 of these loci were observed between zebu and taurine cattle. These markers subsequently proved very useful for investigations of gene flow and admixture in African populations. When these data were considered in conjunction with previous mitochondrial and Y chromosomal studies, a distinctive male-mediated pattern of zebu genetic introgression was revealed. The introgression of zebu-specific alleles in African cattle afforded a high resolution perspective on the hybrid nature of African cattle populations and also suggested that certain West African populations of valuable disease-tolerant taurine cattle are under threat of genetic absorption by migrating zebu herds. PMID- 9215910 TI - The new RGA locus encodes a negative regulator of gibberellin response in Arabidopsis thaliana. AB - We have identified a new locus involved in gibberellin (GA) signal transduction by screening for suppressors of the Arabidopsis thaliana GA biosynthetic mutant gal-3. The locus is named RGA for repressor of gal-3. Based on the recessive phenotype of the digenic rga/gal-3 mutant, the wild-type gene product of RGA is probably a negative regulator of GA responses. Our screen for suppressors of gal 3 identified 17 mutant alleles of RGA as well as 10 new mutant alleles at the previously identified SPY locus. The digenic (double homozygous) rga/gal-3 mutants are able to partially repress several defects of gal-3 including stem growth, leaf abaxial trichome initiation, flowering time, and apical dominance. The phenotype of the trigenic mutant (triple homozygous) rga/spy/gal-3 shows that rga and spy have additive effects regulating flowering time, abaxial leaf trichome initiation and apical dominance. This trigenic mutant is similar to wild type with respect to each of these developmental events. Because rga/spy/gal-3 is almost insensitive to GA for hypocotyl growth and its bolting stem is taller than the wild-type plant, the combined effects of the rga and spy mutations appear to allow GA-independent stem growth. Our studies indicate that RGA lies on a separate branch of the GA signal transduction pathway from SPY, which leads us to propose a modified model of the GA response pathway. PMID- 9215911 TI - Region-specific cis- and trans-acting factors contribute to genetic variability in meiotic recombination in maize. AB - Understanding the genetic basis for variability in recombination rates is important for general genetic studies and plant-breeding efforts. Earlier studies had suggested increased recombination frequencies in particular F2 populations derived from the maize inbred A188. A detailed phenotypic and molecular analysis was undertaken to extend these observations and dissect the responsible factors. A heritable increase in recombination in the sh1-bz1 interval was observed in these populations. A factor causing an approximate twofold increase mapped to the A188 sh1-Bz1 region, behaved as a dominant, cis-acting factor, affected recombination equally in male and female sporogenesis and did not reduce the well studied complete interference in the adjacent bz1-wx interval. This factor also did not increase recombination frequencies in the c1-sh1 and bz1-wx intervals, demonstrating independent control of recombination in adjacent intervals. Additional phenotypic analysis of recombination in the c1-sh1 and bz1-wx intervals and RFLP analysis of recombination along chromosomes 7 and 5 suggested that heritable factors controlling recombination in these intervals act largely independently and in trans. Our results show that recombination in these populations, and possibly maize in general, is controlled by both cis- and trans acting factors that affect specific chromosomal regions. PMID- 9215912 TI - Quantitative trait loci differentiating the outbreeding Mimulus guttatus from the inbreeding M. platycalyx. AB - Theoretical predictions about the evolution of selfing depend on the genetic architecture of loci controlling selfing (monogenic vs. polygenic determination, large vs. small effect of alleles, dominance vs. recessiveness), and studies of such architecture are lacking. We inferred the genetic basis of mating system differences between the outbreeding Mimulus guttatus and the inbreeding M. platycalyx by quantitative trait locus (QTL) mapping using random amplified polymorphic DNA and isozyme markers. One to three QTL were detected for each of five mating system characters, and each QTL explained 7.6-28.6% of the phenotypic variance. Taken together, QTL accounted for up to 38% of the variation in mating system characters, and a large proportion of variation was unaccounted for. Inferred QTL often affected more than one trait, contributing to the genetic correlation between those traits. These results are consistent with the hypothesis that quantitative variation in plant mating system characters is primarily controlled by loci with small effect. PMID- 9215913 TI - Comparison of flowering time genes in Brassica rapa, B. napus and Arabidopsis thaliana. AB - The major difference between annual and biennial cultivars of oilseed Brassica napus and B. rapa is conferred by genes controlling vernalization-responsive flowering time. These genes were compared between the species by aligning the map positions of flowering time quantitative trait loci (QTLs) detected in a segregating population of each species. The results suggest that two major QTLs identified in B. rapa correspond to two major QTLs identified in B. napus. Since B. rapa is one of the hypothesized diploid parents of the amphidiploid B. napus, the vernalization requirement of B. napus probably originated from B. rapa. Brassica genes also were compared to flowering time genes in Arabidopsis thaliana by mapping RFLP loci with the same probes in both B. napus and Arabidopsis. The region containing one pair of Brassica QTLs was collinear with the top of chromosome 5 in A. thaliana where flowering time genes FLC, FY and CO are located. The region containing the second pair of QTLs showed fractured collinearity with several regions of the Arabidopsis genome, including the top of chromosome 4 where FRI is located. Thus, these Brassica genes may correspond to two genes (FLC and FRI) that regulate flowering time in the latest flowering ecotypes of Arabidopsis. PMID- 9215914 TI - Cloning of the Arabidopsis and rice formaldehyde dehydrogenase genes: implications for the origin of plant ADH enzymes. AB - This article reports the cloning of the genes encoding the Arabidopsis and rice class III ADH enzymes, members of the alcohol dehydrogenase or medium chain reductase/dehydrogenase superfamily of proteins with glutathione-dependent formaldehyde dehydrogenase activity (GSH-FDH). Both genes contain eight introns in exactly the same positions, and these positions are conserved in plant ethanol active Adh genes (class P). These data provide further evidence that plant class P genes have evolved from class III genes by gene duplication and acquisition of new substrate specificities. The position of introns and similarities in the nucleic acid and amino acid sequences of the different classes of ADH enzymes in plants and humans suggest that plant and animal class III enzymes diverged before they duplicated to give rise to plant and animal ethanol-active ADH enzymes. Plant class P ADH enzymes have gained substrate specificities and evolved promoters with different expression properties, in keeping with their metabolic function as part of the alcohol fermentation pathway. PMID- 9215915 TI - The maize transposable element Ac induces recombination between the donor site and an homologous ectopic sequence. AB - The prominent repair mechanism of DNA double-strand breaks formed upon excision of the maize Ac transposable element is via nonhomologous end joining. In this work we have studied the role of homologous recombination as an additional repair pathway. To this end, we developed an assay whereby beta-Glucuronidase (GUS) activity is restored upon recombination between two homologous ectopic (nonallelic) sequences in transgenic tobacco plants. One of the recombination partners carried a deletion at the 5' end of GUS and an Ac or a Ds element inserted at the deletion site. The other partner carried an intact 5' end of the GUS open reading frame and had a deletion at the 3' end of the gene. Based on GUS reactivation data, we found that the excision of Ac induced recombination between ectopic sequences by at least two orders of magnitude. Recombination events, visualized by blue staining, were detected in seedlings, in pollen and in protoplasts. DNA fragments corresponding to recombination events were recovered exclusively in crosses with Ac-carrying plants, providing physical evidence for Ac-induced ectopic recombination. The occurrence of ectopic recombination following double-strand breaks is a potentially important factor in plant genome evolution. PMID- 9215916 TI - A comparison of population differentiation across four classes of gene marker in limber pine (Pinus flexilis James). AB - We examined genetic differentiation among seven populations of limber pine using four classes of gene marker. Among-population differentiation was much higher for maternally inherited mitochondrial DNA polymorphisms than for paternally inherited chloroplast DNA, indicating that wind-dispersed pollen is the main agent of gene flow. Chloroplast DNA differentiation is consistent with gene flow estimated in a prior paternity analysis. Using the estimates of seed and pollen flow derived from mtDNA and cpDNA differentiation, we predicted the value of Fst expected at nuclear loci. Allelic frequency differentiation at seven allozyme loci was relatively homogeneous across loci and consistent with the level of differentiation predicted from the organellar haplotypes. By contrast four of the nine randomly applied polymorphic DNA (RAPD) markers we examined were more strongly differentiated than this prediction, suggesting the action of diversifying selection. However, the differentiated RAPDs and mtDNA were concordant in dividing the populations into two groups, suggesting some historical division. Simulations show that such historical division can increase the interlocus variance in Fst, but neither a historical nor an equilibrium model could account for the joint distribution of Fst estimates across both allozyme and RAPD loci. Thus at least one group of loci appears to be experiencing natural selection. PMID- 9215917 TI - The exact test for cytonuclear disequilibria. AB - We extend the analysis of the statistical properties of cytonuclear disequilibria in two major ways. First, we develop the asymptotic sampling theory for the nonrandom associations between the alleles at a haploid cytoplasmic locus and the alleles and genotypes at a diploid nuclear locus, when there are an arbitrary number of alleles at each marker. This includes the derivation of the maximum likelihood estimators and their sampling variances for each disequilibrium measure, together with simple tests of the null hypothesis of no disequilibrium. In addition to these new asymptotic tests, we provide the first implementation of Fisher's exact test for the genotypic cytonuclear disequilibria and some approximations of the exact test. We also outline an exact test for allelic cytonuclear disequilibria in multiallelic systems. An exact test should be used for data sets when either the marginal frequencies are extreme or the sample size is small. The utility of this new sampling theory is illustrated through applications to recent nuclear-mtDNA and nuclear-cpDNA data sets. The results also apply to population surveys of nuclear loci in conjunction with markers in cytoplasmically inherited microorganisms. PMID- 9215918 TI - A population genetics model of marker-assisted selection. AB - A deterministic two-loci model was developed to predict genetic response to marker-assisted selection (MAS) in one generation and in multiple generations. Formulas were derived to relate linkage disequilibrium in a population to the proportion of additive genetic variance used by MAS, and in turn to an extra improvement in genetic response over phenotypic selection. Predictions of the response were compared to those predicted by using an infinite-loci model and the factors affecting efficiency of MAS were examined. Theoretical analyses of the present study revealed the nonlinearity between the selection intensity and genetic response in MAS. In addition to the heritability of the trait and the proportion of the marker-associated genetic variance, the frequencies of the selectively favorable alleles at the two loci, one marker and one quantitative trait locus, were found to play an important role in determining both the short- and long-term efficiencies of MAS. The evolution of linkage disequilibrium and thus the genetic response over several generations were predicted theoretically and examined by simulation. MAS dissipated the disequilibrium more quickly than drift alone. In some cases studied, the rate of dissipation was as large as that to be expected in the circumstance where the true recombination fraction was increased by three times and selection was absent. PMID- 9215919 TI - The coalescent process with selfing. AB - A method of estimating the selfing rate using DNA sequence data was recently proposed by Milligan. Unfortunately, a number of errors make interpretation of his results problematic. In the present paper we first show how the usual coalescent process can be adapted to models that include selfing, and then use this result to find moment estimators as well as the likelihood surface for the selfing rate, s, and the scaled mutation rate, theta. We conclude that, regardless of the method used, large sample sizes are necessary to estimate s with any degree of certainty, and that the estimate is always highly sensitive to recent changes in the true value. PMID- 9215920 TI - A test of neutrality based on interlocus associations. AB - The evolutionary processes governing variability within genomic regions of low recombination have been the focus of many studies. Here, I investigate the statistical properties of a measure of interlocus genetic associations under the assumption that mutations are selectively neutral and sites are completely linked. This measure, denoted ZnS, is based on the squared correlation of allelic identity at pairs of polymorphic sites. Upper bounds for ZnS are determined by simulations. Various deviations from the neutral model, including several different forms of natural selection, will inflate the value of ZnS relative to its neutral theory expectations. Larger than expected values of ZnS are observed in genetic samples from the yellow-ac-scute and Adh regions of Drosophila melanogaster. PMID- 9215921 TI - Element specification in biological monitoring. PMID- 9215922 TI - Critical evaluation and review of cadmium concentrations in blood for use in occupational health according to the TRACY protocol. AB - Cadmium in blood (B-Cd) may be used to assess recent exposure to cadmium in the working or general environment. In a paper published elsewhere pooled reference values using meta-analysis of B-Cd values in general-population studies were calculated. In the present study tentative references intervals were described which can be used for comparison with data from occupationally exposed groups or individuals. The selection of studies was done according to criteria as published by the international project TRACY. For this purpose, 800 publications covering the period 1983-1992 were reviewed on their suitability for establishing tentative reference intervals. From these 800 publications, four finally met the selection criteria. Most important criteria for selection were the check for contamination during sampling of the blood, the storage and pretreatment procedures, and the existence of internal and external quality control programs. Also, stratifications into sex, smoking habits and occupation were important selection criteria. It turned out that for non-smoking white-collar workers in the age range of 19-65 years, B-Cd values were below 0.8 micrograms/l for most areas. All other groups within this age group, e.g., white collar workers in Japan, blue-collar workers, and smokers tend to have higher B-Cd values in these sequences. Blue-collar workers not clearly exposed to Cd have higher values than white-collar workers, indicating still some minor exposure. It is not clear if this small exposure has an occupational or lifestyle (e.g., diet) origin. Geographical regions also show an influence on B-Cd levels, e.g., values in Japan are higher than elsewhere. This influence may be due to differences in diet. The conclusion will be that reference values for B-Cd in fact are area-dependent. PMID- 9215923 TI - The integrity of the liver among people environmentally exposed to cadmium at various levels. AB - The objective of the present study was to examine if environmental exposure to cadmium (Cd) had any impact on the integrity of the liver among the general Japanese population. A nationwide survey was conducted in the winter seasons of 1991-1995 in 15 prefectures in Japan to collect 24-h food duplicates, peripheral blood samples, and morning spot urine samples from healthy nonsmoking adult women. The samples were analyzed for Cd by automated graphite furnace atomic absorption spectrometry. In total, 371 women offered food duplicate, blood, and urine samples. The dietary Cd intake was 17.3-79.4 micrograms/day, the level of Cd in blood was 1.58-3.82 ng/ml, and the urinary Cd concentration was 1.06-4.74 micrograms/g creatinine as geometric means calculated on a regional group basis. Analyses for correlation with liver function showed no Cd-exposure-related elevation in enzyme levels or reduction in albumin levels' in serum. The distribution of cases with enzyme levels above normal ranges (below normal in the case of albumin) did not show any dose-related bias. The age of the subject, not the exposure to Cd, was the most influential factor in determining serum enzyme levels. Environmental exposure to Cd has not affected the integrity of the liver among the general population in Japan. PMID- 9215924 TI - The epidemiology and surveillance of blood lead in Taiwan (ROC): a report on the PRESS-BLL project. AB - To monitor the lead hazards in industries and to investigate the prevalence of elevated blood lead levels (BLLs) in lead-exposed workers, a lead surveillance system (PRESS-BLLs) has been established and operated in Taiwan, Republic of China, since July 1993. A cohort of lead-exposed workers who received a periodic annual health examination at 55 accredited hospital laboratories was constructed. A total of 9807 separate BLL measurements were reported to the system in 1994. The mean BLL was 15.8 micrograms/dl in male workers and 11.6 micrograms/dl in female workers. The mean BLL of lead-exposed workers was significantly (P < 0.05, z-test) higher than that of the general Taiwanese population (8.6 micrograms/dl for males and 6.7 micrograms/dl for females). In addition, the BLLs of 983 (10.0%) workers exceeded the regulatory action level (40 micrograms/dl for males; 30 micrograms/dl for females). The workplaces and homes of 57% of the workers with elevated BLLs were thoroughly investigated to determine the sources of lead contamination. These actions identified the causes of elevated BLLs and set up strategies to reduce workers' lead exposure. The establishment of this occupational lead surveillance system represents a method for monitoring of lead hazards from occupational and environmental settings to prevent lead poisoning. The information acquired from the system can help in the setting up of a priority of prevention and the development of control measures. It is also useful for further monitoring of changes in the BLLs of the lead-exposed cohort. The Health Department of Taiwan can use this information to evaluate the effectiveness of current industrial hygiene practice. Subjects with elevated BLLs have been medically treated and placed on long-term follow-up for sequelae. PMID- 9215925 TI - Symptoms, lung and liver function, blood counts, and genotoxic effects in coastal tanker crews. AB - OBJECTIVE: The deck crew on tankers can be exposed to high concentrations of benzene and other chemicals during loading, unloading and tank-cleaning operations. The objective of this study was to investigate whether genotoxic or other early health effects of cargo vapour exposure could be detected in coastal tanker crews. METHODS: The association between exposure to cargo vapours and clinical symptoms and signs, spirometry, blood cell count, blood test for liver function, and the frequency of micronuclei and sister chromatid exchanges in peripheral lymphocytes was studied in a cross-sectional investigation of 107 male crew members (66 deck crew and 41 others) on ten coastal tankers. RESULTS: Seven of the tankers had automatic cargo level gauging systems but some of the ships still had open hatches during loading and unloading operations. Acute symptoms such as headache, nausea, vertigo, fatigue and dizziness after loading or tank cleaning operations were reported by 56 of the 66 deck crew members (85%). Irritation of the mucous membrane in eyes and upper respiratory tract by cargo vapours were also common in this group. Obstructive symptoms were more common in the group with the highest exposure to cargo vapours but persistent effects on lung function (vital capacity and forced expiratory volume in 1 s), nervous system, liver enzymes or blood counts were not found. The frequency of micronuclei after mitotic stimulation with phytohaemagglutinin was higher among the deck crew (mean 4.2 SEM 0.40) than in other crew members (mean 3.6, SEM 0.35). although the difference was not statistically significant. We found no association between exposure and the frequency of sister chromatid exchanges or micronuclei after stimulation with pokeweed mitogen. CONCLUSION: This study indicates that exposure to cargo vapours in coastal tanker crews may cause symptoms in the respiratory and nervous systems. PMID- 9215926 TI - Internal exposure to hazardous substances of persons from various continents: investigations on exposure to different organochlorine compounds. AB - The aim of the study was to investigate the concentration of organochlorine compounds of environmental-medical relevance in biological materials from refugees with regard to their countries of origin and to compare these concentrations with the internal exposure of the German general population. METHODS: During medical examination after entry to Germany specimens could be taken from the refugees to determine the following parameters of biological monitoring: 1,1-dichloro-2,2-bis(-chlorophenyl)-ethylene (DDE-P), polychlorinated biphenyls (PCB-P), pentachlorophenol (PCP-P) and the beta- and gamma hexachlorocyclohexanes (beta-HCH-P, gamma-HCH-P) in plasma and the excretion of chlorophenols (4-MCP-U, 2,4-DCP-U, 2,5-DCP-U, 2,4,5-TCP-U, 2,4,6-TCP-U, 2,3,4,5 TeCP-U, 2,3,5,6-TeCP-U) in urine. One hundred and three men (13 from former Yugoslavia, 29 from the former USSR, 33 Africans and 28 Asians) ranging from 16 to 53 years of age (median 27 years) were investigated. Thirty four male Germans without occupational exposure to these substances and a similar age structure (age 25-36 years; median 26 years) served as a control group. RESULTS: A much higher level of internal exposure was found for the DDT metabolite, DDE, for those persons from Asia, the former USSR and Africa compared with the German controls (medians: 16.9 micrograms/l, 11.9 micrograms/l and 10.9 micrograms/l) and 1.1 micrograms/l). The levels of PCB in plasma were below the detection limit in the majority of refugees. In the control group, however, the PCB levels were higher (sigma PCB; median: 2.1 micrograms/l, maximum: 13.3 micrograms/l). The highest beta-HCH concentrations, up to a maximum of 15.5 micrograms/l, were detected in the persons from the former USSR and Asia. The five groups do not differ with regard to internal exposure to PCP and gamma-HCH. Renal excretion of 4-MCP, 2,4-DCP and TeCP did not differ between the five groups. The concentrations of 2,5-DCP in urine, however, were significantly lower in the Germans than the refugees from the four regions investigated. The median for the Germans was 3.0 micrograms/l and for the refugees between 10.8 and 14.7 micrograms/l. Also the levels of 2,4,5-TCP and 2,4,6-TCP in urine were lower in the German controls than in the men from the former USSR, Africa and Asia. CONCLUSIONS: Organochlorine compounds exist worldwide due to their extensive use. There are, however, regional differences for the various substance groups, which during biological monitoring are seen in the different background exposures of the general population. Particularly characteristic are markedly higher levels of DDE in plasma from the refugees compared with the German population. PMID- 9215927 TI - The influence of cognitive bias on the perceived odor, irritation and health symptoms from chemical exposure. AB - OBJECTIVE: Responses to volatile chemicals are often subjective and variable, both over time and across individuals. Although variability can derive from differences in individual olfactory sensitivity, the response to a chemical stimulus is also influenced by the complex environment surrounding the exposure, which can include the perceiver's cognitive state. To explore the role of cognitive bias in chemical exposures, we evaluated whether information about the consequences of exposure to acetone could influence ratings of odor and irritation during exposure and/or the frequency or intensity of reported health symptoms following exposure. METHODS: Ninety adults (mean age 33.7, range 25-64) with no history of occupational exposure to solvents, were exposed to 800 ppm acetone in a chamber for 20 min. To control for non-specific responses to the odor of acetone, the subjects were also exposed for 20 min to 200 ppm phenylethyl alcohol (PEA), a non-irritant volatile chemical that produces a distinct odor but does not elicit irritation in the vapor phase. Subjects were assigned to one of three groups (n = 30/group); each group was given either a positive, negative or neutral bias towards the consequences of exposure to the chemicals in the study. During exposure, subjects rated the intensity of odor and irritation; following exposure, they completed symptom questionnaires. RESULTS: During the 20-min exposure to acetone, the positive bias group exhibited the most adaptation to its odor and the lowest perceived irritation; following exposure they reported the fewest health symptoms. In contrast, the negative bias group rated higher levels of odor intensity and, on average, reported the most over-all irritation; following exposure they reported significantly more health symptoms than the other groups, None of the demographic variables studied (e.g., age, gender, race, smoking status) were predictive of the response to odor or irritation. The perceived irritancy of acetone was well predicted by a linear combination of the perceived odor of acetone and perceived irritation for PEA (the nonirritant), r2 = 0.73. CONCLUSIONS: The results provide strong evidence that both the perceived odor and cognitive expectations about a chemical can significantly affect how individuals respond to it. Moreover, because naive control subjects appear to exhibit extreme variation in their cognitive evaluations of chemical effects, there may be limited value in using non-exposed controls to assess the irritancy of chemicals for worker populations. PMID- 9215928 TI - Proposal for hand-arm vibration exposure limits adopted for Japanese workers operating hand-held vibration tools. AB - On the basis of data presented in our previous, reports, the current study was undertaken to estimate frequency-weighted hand-arm vibration exposure limits for various daily exposure times. The procedures for the present study were as follows. (1) The prevalence of vibration-induced white finger (VWF) as well as the vibration exposure were investigated in various groups of workers operating hand-held vibrating tools. The vibration magnitude of various tools was measured and the results were presented as the energy-equivalent frequency-weighted root mean-square (m/s2 rms) acceleration. There was a statistically significant positive correlation between the prevalence of VWF and the measured vibration magnitude (R2 = 0.5, P < 0.05). Hence, it was concluded that in decisions concerning quantitative recommendations for vibration exposure, the prevalence of VWF should be considered. (2) By a careful selection of available publications which contain useful information on duration of vibration exposure of < or = 2 h/day and the occurrence of VWF, a significant correlation between the prevalence of VWF and the vibration magnitude could be observed. The regression equation was estimated as: y = -18.5 + 4.6 (x), R2 = 0.8. On the basis of this equation, it was speculated that the prevalence of VWF in workers using vibrating tools might be restricted to the prevalence of Raynaud's phenomenon in the Japanese general population if the 2-h daily vibration exposure is about 4.5 m/s2 rms. (3) Regarding this speculation, the equation provided in the documentation of ISO 5349 was used and modified as: [alpha h,w)eq,t = (alpha h,w)eq,2(2/t)1/2 (m/s2 rms)] and then the vibration limit values for daily exposure of 1 min to 8 h were calculated. (4) In order to achieve compatibility with standards of other countries, and to formulate an easy method for using the recommended values presented here, the daily exposure time of 8, 4, 2, 1 and 0.5 h were selected. The correspondence vibration magnitudes were in the range 22-9.0 m/s2 rms, and the lower limit (2.2 m/s2 rms) was assumed as the permissible vibration exposure limit for an 8-h working period. The proposed daily vibration limits were then compared with those recommended by other institutions. PMID- 9215929 TI - Load pattern and pressure pain threshold in the upper trapezius muscle and psychosocial factors in medical secretaries with and without shoulder/neck disorders. AB - OBJECT: A current hypothesis for the genesis of muscular complaints in the shoulder/neck region postulates that short periods with a completely relaxed muscle are essential to avoid complaints. Another hypothesis is that these disorders are related to psychosocial conditions at work. In order to test these hypotheses, 23 medical secretaries were investigated. METHODS: The load pattern during work in the upper trapezius muscle bilaterally was assessed with electromyographic (EMG) technique and exposure variation analysis (EVA). In addition, pressure pain threshold (PPT) was measured on the trapezius muscle bilaterally and on the sternum. Psychosocial conditions at work were assessed with a questionnaire. RESULTS: The medical secretaries with complaints had significantly fewer episodes with totally or close to totally relaxed muscle compared with the healthy group. The group with complaints tended to have a more monotonous load pattern at low levels (approx. 1%-5% maximum voluntary contraction) while the healthy group had more frequent pauses but also somewhat more frequent short load peaks. The group with complaints showed lower PPT readings compared with the healthy group. However, the whole group had considerably lower PPTs than is usually reported in the literature. Of the 12 questions in the psychosocial questionnaire only one regarding work task satisfaction showed a significant difference between the two groups. CONCLUSION: Support is found for hypothesis that secretaries without complaints have more frequent episodes with totally relaxed muscle. A significant difference is found regarding work task satisfaction. PMID- 9215930 TI - Combined effects of hand-arm vibration and noise on temporary threshold shifts of hearing in healthy subjects. AB - OBJECT: To investigate whether hand-arm vibration and noise have a combined effect on temporary threshold shift (TTS) of hearing among healthy subjects. METHOD AND DESIGN: Nineteen healthy subjects with an average age of 25.7 (SD 7.7) years were exposed to vibration (30 m/s2, 60 Hz), noise [90 dB(A)] and both, respectively. The subject's right hand was placed on the plate of a vibrator and the right ear exposed to noise via headphones. Subjects were exposed to vibration and/or noise for 3 min and after a 1-min pause the exposure was repeated five times. Hearing thresholds at 1, 4 and 6 kHz were measured during the time periods before, between (during pauses) and after exposure. RESULTS: Exposure to vibration alone caused almost no hearing threshold changes at every frequency tested. But exposure to noise or a combination of vibration and noise caused a significant increase in TTSs at 4 and 6 kHz. Moreover, exposure to a combination of vibration and noise caused significantly higher TTSs than exposure to noise at 4 and 6 kHz. CONCLUSION: The present results demonstrate the combined effects of hand-arm vibration and noise can enhance exposure to hand-arm vibration and noise can enhance the TTS of hearing more than noise exposure, though hand-arm vibration alone may hardly affect TTS. PMID- 9215931 TI - The influence of biodynamic factors on the mechanical impedance of the hand and arm. AB - The purpose of this study was to investigate the mechanical impedance of the human hand-arm system during exposure to random vibration under various experimental conditions and to evaluate statistically whether these experimental conditions have any influence on magnitude and phase of the mechanical impedance. A further aim was to compare the obtained results with other investigations where sinusoidal excitation has been used. The mechanical impedance was estimated in ten healthy subjects during exposure to random vibration, with a constant velocity spectrum within the frequency range 4-2000 Hz, by use of a specially designed laboratory handle. In the study, the influence of various conditions, such as vibration direction (Xh, Yh, Zh), grip force (25-75 N), feed force (20-60 N), frequency-weighted acceleration level (3, 6, 9, 12 m/s2) and hand and arm posture (five flexions, two abductions) were studied. The outcome showed that the vibration direction and the frequency of the vibration stimuli have a strong significant influence on the impedance of the hand. An increased vibration level resulted in a significantly lower impedance for frequencies over 100 Hz. Increase grip and feed forced led on the other hand to an increased impedance for all frequencies. With regard to hand and arm posture, the results show that the flexion and abduction had a significant contribution for frequencies below 30 Hz. Furthermore, the influence of some of the studied variables had a non-linear effect on the impedance but also differed between different exposure directions. It was concluded, moreover, that the vibration response characteristics of the hand and arm differ, depending whether the signal is a discrete frequency signal or a signal consisting of several frequencies. PMID- 9215932 TI - Effects of obesity, current smoking status, and alcohol consumption on heart rate variability in male white-collar workers. AB - In order to examine the effects of mild to moderate obesity, moderate to heavy smoking, and moderate alcohol consumption on cardiac parasympathetic activities and systemic sympathetic activities, a cross-sectional survey was carried out in 282 healthy Japanese male white-collar workers. Their autonomic activities were assessed as amplitudes of spectral components of heart rate variability (HRV) which was measured in the annual physical examination at their work sites. Taking the effects of aging on HRV into account, the cardiac parasympathetic activity at supine rest and its response to a change in posture were reduced in mildly to moderately obese subjects with a body mass index of 21-36, whereas the sympathetic activity was not. The effects of smoking and alcohol consumption on HRV were not confirmed. The above results means that we should consider obesity as a covariate when we examine possible relationships between cardiac parasympathetic activity and other environmental factors. There is a need for further studies on the relationships among obesity, change in parasympathetic activity, and development of health problems. The dose-effect relationships between long-term smoking or alcohol consumption and chronic changes in autonomic activities also remain to be determined. PMID- 9215933 TI - Biological monitoring of styrene exposure and possible interference of acetone co exposure. AB - The object of this study is the evaluation of some of the toxicokinetic effects of exposure to low concentrations of styrene, and the possible influence of simultaneous exposure to acetone. To this end we studied 19 workmen simultaneously exposed to both solvents. During a week of 4-h work shifts, the workmen underwent daily personal environmental monitoring and the collection of urine samples, at both the beginning and the end of the work period, for the determination of mandelic acid (MA) and phenylglyoxylic acid (PGA). The presence of the solvents in the atmosphere was evaluated using passive personal monitoring and gas chromatography. Average exposure to styrene and acetone were respectively 72.2 mg/m3 and 225.7 mg/m3. MA and PGA were quantified by high-performance liquid chromatography (HPLC). The daily urinary concentration averages, both at commencement and at the end of work shifts, of both the metabolites studied and of the sum of the two were in statistically significant linear correlation with the average daily styrene exposure. Concentrations of MA and PGA in urine samples collected at the start of the work shift averaged 61.5 mg/g creatinine and 45.2 mg/g creatinine respectively, representing 41% and 72% of those at the endo of the work shift which were 148.3 and 62.6 mg/g creatinine, respectively. With equal exposure to styrene, the average urinary concentrations of MA and PGA at both the beginning and end of the work shift increased significantly (P < 0.001) during the working week. Moreover, we found that with equal exposure to styrene, urinary excretion of MA, PGA and MA + PGA at the end of the shift was inversely correlated with the intensity of acetone exposure (r = 0.4659, 0.3410 and 0.542 respectively, P < 0.001). In conclusion, these results express slower urinary kinetics of styrene metabolites than is usually described in the literature, and favor a tendency to accumulate MA and PGA in the organism as a consequence of the retardation of urinary excretion kinetics. Acetone apparently represents one of the determining factors in this interference. PMID- 9215934 TI - Quantifying work load in neck, shoulders and wrists in female dentists. AB - OBJECTIVE: To assess the work load in neck and upper limbs of dentists. METHODS: Twelve right-handed female dentists (six with and six without a history of definite neck/shoulder disorders, pair-wise matched for age) were studied when performing authentic dental work. Electromyography (EMG) was used to quantify the muscular load of the shoulders bilaterally and of the right forearm. Positions and movements of the head and wrists were measured, using inclinometers and electrogoniometers. RESULTS: During work, the median load for the right upper trapezium muscle was 8.4% of the maximal voluntary EMG activity (MVE); during 90% of the time the load was > or = 3.3% MVE ("static" load). The figures were somewhat lower on the left side (7.0% and 2.5% MVE, respectively). Subjects with disorders had over all lower load levels for the trapezius muscles, although not statistically significant at < 0.05, than those without disorders. During a standardized reference contraction for the trapezius, the load was 17% MVE, and the quotient between MVE and torque [normalized to maximal voluntary torque (MVC)] was 0.5. These figures may be used for transformations. The muscular load on the right forearm was similar to the loads on the trapezius. The head was, on average, forward tilted > or = to 39 degrees, and during 10% of the time > or = 49 degrees. The left hand was held in more static positions, with palmar flexion and ulnar deviation, also reflected by lower angular velocities and repetitiveness, as compared with the right one, which was dorsiflexed. CONCLUSIONS: Dentists are exposed to high load on the trapezius muscles bilaterally, and steep, prolonged forward bending of the head. Further, for the wrists the postures were constrained, but the dynamic demands were low. PMID- 9215935 TI - Evaluation of the effects of an ergonomic-educational programme. The assessment of "ergonomic errors" made during the performance of nursing tasks. AB - OBJECTIVES: The objectives of this study were (i) To establish whether it is possible to assess by means of a check-list in a reliable way errors which violated biomechanical and ergonomical principles during nursing tasks, and (ii) to study the effectiveness of an ergonomic-educational course by using this check list. MATERIALS AND METHODS: Trainees (n = 12) and a control group of nurses (n = 12) who did not attend the course, performed three nursing activities at three points in time under standardized conditions; once before and twice after the course had ended. Their performances were recorded on video. A check-list was developed to assess the number of ergonomic errors made during the test performances. Two observers completed the check-lists after having watched five videotapes, and one of them did this for a second time 3 weeks later, in order to assess inter- and intra-observer reliability. In addition the tapes of all nurses were scored and analysed on differences in the performance of the two groups at the three points in time. Percentage of agreement and kappa (kappa) was used to express inter- and intra-observer reliability. Student's t-test was used to analyse the differences in mean percentages of errors. RESULTS: The inter- and intra-observer reliability were 92% with kappa of 0.84 and 93% with kappa of 0.86, respectively. Further results showed that the mean percentages of errors made by the control group remained the same at the three measurement times. However, in the trainee group a significant decrease in errors was found. The trainees made fewer errors at all three points in time than the controls did. CONCLUSION: It appears feasible to create a check-list to assess ergonomic errors in a reliable way. Trainees make fewer errors after an ergonomic-educational course. It is necessary, however to evaluate whether trained nurses work more safely in their daily duties than during the study. PMID- 9215936 TI - Computer-assisted classification system for chest X-ray and computed tomography findings in occupational lung disease. AB - The ILO classification of pneumoconiotic changes in the lungs and pleura has become a standardized and widely accepted method of documentation in occupational medicine. Recently a classification system for computed tomography/high resolution computed tomography (CT/HRCT) findings in the lung has been proposed as well. For both classification systems, computed-assisted programs have been developed that allow the storage and archiving of the results as well as further statistical processing. The programs are compatible with usual hardware configurations, have a comprehensible and transparent structure, and are easy to learn and adaptable to individual needs. The use of the computer-assisted classification systems is presented in the example of an insulator exposed to asbestos. The system of data documentation and processing has proved to be very practicable in the more than 2000 patients studied thus far. PMID- 9215937 TI - The risk of occupational rhinitis. AB - OBJECTIVE: Reports on the aetiology and risk of occupational rhinitis in different occupations are scarce. METHOD: The purpose of this study was to find the occupations with an increased risk of occupational rhinitis. Age and gender differences in occupational rhinitis and occupational asthma were also compared, and the most common causes of occupational rhinitis were analysed. DESIGN: This study covered the cases of occupational rhinitis and asthma reported to the Finnish Register of Occupational Diseases during the years 1986-1991. The cases on the Register were linked to the longitudinal census data file from the Finnish censuses. RESULTS: During 1986-1991, 1244 new cases of occupational rhinitis (474 women and 497 men) and 1867 new cases of occupational asthma (916 women and 951 men) were reported to the Register. Animal dander, flours, wood dusts, textiles, phthalic acid anhydrides and storage mites were important causes of occupational rhinitis. The highest relative risk of occupational rhinitis was among furriers, the age-standardized rate ratio (SRR) was 30.0. Bakers and livestock breeders had also a markedly elevated relative risk (SRR = 22.0). Men had the highest incidence of occupational rhinitis at the age of 25-29 years and among women the incidence gradually increased and reached the peak in the group 40-44 years of age. CONCLUSION: Furriers, bakers, and livestock breeders had the most elevated relative risk of occupational rhinitis. Occupational rhinitis cases reported at a younger age than asthma, suggesting that rhinitis often precedes asthma. PMID- 9215938 TI - Urinary excretion of dimethylhippuric acids in humans after exposure to trimethylbenzenes. AB - The aim of this study was to determine the urinary excretion of dimethylhippuric acids (DMHAs) in humans after experimental chamber exposure to trimethylbenzene (TMB) vapor. The DMHAs have been put forward as suitable biomarkers of exposure to products containing TMBs such as white spirit and petrol. Ten healthy male volunteers were exposed to TMB vapor in an exposure chamber for 2 h at a work load of 50 W. The subjects were exposed on four occasions, to 25 ppm of 1,2,4 TMB, 1,2,3-TMB, and 1,3,5-TMB, respectively, and 2 ppm of 1,2,4-TMB. Urine was collected from the onset of exposure until the following morning. All six possible DMHA isomers were analyzed by high-performance liquid chromatography. About 22% of the inhaled amount of 1,2,4-TMB was excreted as DMHAs within 24 h, mainly as 3,4-DMHA. The 24-h recovery of 1,2,3-TMB as DMHAs was 11%. Only 3% of the absorbed amount of 1,3,5-TMB was excreted as 3,5,-DMHA. The half-times of the different DMHA isomers ranged from 4 to 16 h. In addition to analysis of DMHAs, the excretion of unconjugated dimethylbenzoic acids in urine was estimated to account for approximately 3% of the dose of all TMBs. In conclusion, the urinary excretion of DMHA isomers may serve as a good indicator of TMB exposure. In this controlled short-term-exposure study the sum of excretion rate of several DMHA isomers reflected exposure more closely than did the excretion rate of any single DMHA. PMID- 9215939 TI - A study of the quality of life in elderly people using psychological testing. AB - To study factors which influence the quality of life (QOL) in the elderly, we investigated the relationship between scores on the modified Philadelphia Geriatric Center (JPGC) Morale Scale and various other psychological tests in 51 elderly people residing in a long-term care facility. The JPGC Morale Scale score correlated with the scores for all sections of the Japanese version of the Cornell Medical Index (JCMI), but not with those for the Mini Mental State Examination, the Kohs block design test, the Bender Gestalt test and the activities of daily living (ADL). Both somatic and psychotic symptoms on the JCMI were correlated with the dementia behaviour disturbance scale score and walking ability according to the ADL. Subjects were further divided into four groups according to Fukamachi's neurotic discriminative diagram based on the JCMI. Scores for most sections of somatic and psychotic symptoms on the JCMI were elevated in proportion to the degree of neurotic tendencies in the elderly. These results indicate that the QOL of the elderly is influenced by subjective symptoms, but not by the degree of cognitive impairment or the ADL. PMID- 9215940 TI - Outcome of delirium: Part 1. Outcome of delirium diagnosed by DSM-III-R, ICD-10 and CAMDEX and derivation of the Reversible Cognitive Dysfunction Scale among acute geriatric inpatients. AB - OBJECTIVE: To study performance of DSM-III-R, ICD-10 and CAMDEX diagnoses of delirium as predictors of improvement in mental state in survivors, and to develop a brief rating scale which will predict reversibility of cognitive dysfunction. DESIGN: Prospective cohort study. SETTING: Acute geriatric inpatient units. PATIENTS: A random sample of consecutive acute admissions of patients over the age of 65 (N = 80). MAIN MEASUREMENTS: Serial assessments of mental state and cognitive function and observational data. Establishment of DSM-III-R, ICD-10, CAMDEX diagnoses. OUTCOME MEASURE: Patients with more than five points of 20% improvement in Mini Mental State Examination following the most severely impaired assessment operationally designated 'reversible cognitive dysfunction'. MAIN RESULTS: Diagnoses of delirium by DSM-III-R and ICD-10 do not predict improvement in cognitive function well; CAMDEX does rather better. Discriminant function analysis yielded the Reversible Cognitive Dysfunction Scale (RCDS), a simple clinical scale which accurately predicted improvement. This comprised reduced conscious level, poor attention, poor contact with the patient, incoherent speech, reduced psychomotor activity, lack of awareness of surroundings and poor orientation and memory. CONCLUSIONS: The concept of and diagnostic criteria for delirium should be reconsidered. The RCDS merits further evaluation. PMID- 9215941 TI - Outcome of delirium: part 2. Clinical features of reversible cognitive dysfunction--are they the same as accepted definitions of delirium? AB - OBJECTIVE: To describe the clinical features of reversible cognitive dysfunction. DESIGN: Prospective cohort study. SETTING: Acute geriatric inpatient units. PATIENTS: A random sample of consecutive acute admissions of patients over the age of 65 (N = 80). MAIN MEASUREMENTS: Serial assessments of mental state and cognitive function and observational data. OUTCOME MEASURE: Patients with more than five points of 20% improvement in Mini Mental State Examination following the most severely impaired assessment operationally designated 'reversible cognitive dysfunction'. The clinical features of those with RCD are compared with those with non-reversible cognitive dysfunction. MAIN RESULTS: Delusions, hallucinations, aggression, excitement, irritability and other 'active' symptoms were not commoner in RCD than in non-reversible cognitive dysfunction (non-RCD). By contrast, 'quiet' signs, such as plucking at bedclothes, poor attention, incoherent speech, abnormal associations, slow, vague thought and fluctuating mental state were more marked in RCD than in non-RCD. CONCLUSIONS: Reversible cognitive dysfunction is a quiet and unobtrusive disorder. PMID- 9215942 TI - The Clock Drawing Test for dementia of the Alzheimer's type: A comparison of three scoring methods in a memory disorders clinic. AB - OBJECTIVES: To examine the reliability and validity of the Clock Drawing Test when used as a cognitive screening instrument for mild to moderate dementia, and to compare different scoring mechanisms. DESIGN: Retrospective analysis of clock drawing performance using three published scoring methods (Shulman, Sunderland and Wolf-Klein). SETTING: Hospital-based memory disorders clinic. PARTICIPANTS: A sample of 28 consecutive patients attending the memory clinic for assessment who were given a diagnosis of Alzheimer's disease (mild or moderate) and 28 age- and sex-matched control subjects comprising 17 memory clinic attenders found to be normal and 11 community volunteers. MEASUREMENTS: Sensitivity and specificity of the three clock rating scales against memory clinic diagnoses of dementia using DSM-III-R; their respective interrater reliabilities; and comparisons of each with measures of cognitive impairment (the Mini-Mental State Examination and the Blessed Orientation-Information-Memory-Concentration Test), daily performance of basic and instrumental activities (the Blessed Dementia Scale) and depression (the Hamilton Rating Scale for Depression). RESULTS: All methods of scoring the Clock Drawing Test correlated well with measures of cognitive impairment (r = 0.57-0.73) and daily performance (r = 0.38-0.48), were independent of mild depression and demonstrated high sensitivity, specificity and interrater reliability. While all clock scales identified mild to moderate dementia reasonably well, the Shulman method performed best. In screening for dementia, clock drawing proved superior to the MMSE: 24/28 vs 20/28 cases identified. When compared with the MMSE, clock drawing provided additional diagnostic discrimination, identifying 7/8 AD patients with MMSE scores = 24. CONCLUSIONS: In a clinic population, clock drawing, especially if scored according to the Shulman scale and combined with the MMSE, is an extremely efficient test screening measure for mild to moderate dementia of the Alzheimer's type with low false negative and false positive rates. This may have implications for screening elderly populations. PMID- 9215943 TI - The relationship between two scales measuring aggressive behaviour among elderly nursing home residents. AB - Data from a large 6-month prospective study of aggressive behaviour in 11 nursing homes for the elderly are used to examine the relationship between two aggression scales (SOAS and RAGE). There was a strong and significant correlation between the SOAS and RAGE scales for the total and the subscale scores. Both these scales appear useful in measuring aggressive behaviour in nursing homes for the elderly and the nursing home staff can be trained in completing them effectively. PMID- 9215944 TI - Elder abuse in people with dementia in Northern Ireland: Prevalence and predictors in cases referred to a psychiatry of old age service. AB - OBJECTIVE: To establish the prevalence of elder abuse in community-dwelling patients with dementia and to test the hypothesis that there is no difference in carer and patient characteristics between the abused and non-abused populations. DESIGN: A cohort of consecutive referrals was formed and subdivided by the presence or absence of abuse and the two groups compared. SETTING: A rural psychiatry of old age service in N. Ireland. SUBJECTS: Each case had been newly referred, was 65 years old or over, lived at home, had an identifiable carer and met DSMIII-R criteria for a diagnosis of dementia. There were 49 such cases; 38 carers agreed to be interviewed. MAIN OUTCOME MEASURES: The General Health Questionnaire 28, the Gilleard Pre-Morbid Relationship Rating Scale and Gilleard's Problem Checklist were administered to the carer and the information/orientation sub scale of the Clifton Assessment Procedure for the Elderly used to measure cognitive impairment in the patient. RESULTS: Abuse was elicited in 14 (37%) cases; four (10.5%) of physical and 13 (34%) of verbal abuse. No cases of abuse by neglect were detected. A poor premorbid relationship, verbal or physical abuse by the dependant, problem behaviours in the dependant, the carer's level of anxiety and a perception of not receiving help were significantly associated with abuse. Alcohol consumption of the carer, physical dependence, severity of cognitive impairment or financial or social circumstances were not associated with elder abuse. CONCLUSIONS: Elder abuse is associated with aspects of the patient/carer relationship and should be regarded as a significant problem in patients with dementia referred to an old age service. PMID- 9215945 TI - Identifying older people with dementia: the effectiveness of a multiservice census. AB - A census of all relevant services in an area can be used to identify people with mental impairment suggestive of dementia. Two censuses in Tayside, Scotland, were used to test the effectiveness of this method. False positives accounted for 12% of returns. After excluding false positives, by comparison with expected dementia prevalence based on EURODEM, 66% of all sufferers and 50% of those living in the community were identified by the censuses. By pro-rating for non-response, the proportion of sufferers known to services was estimated as 72%. The characteristics of those not known to services are unclear and further research is needed on this. The cost of a census in an area of 250,000 population is under pounds 3000. A multiservice census offers a simple, inexpensive, practicable method of constructing a sample frame for population needs assessment. PMID- 9215946 TI - A population needs assessment profile for dementia. AB - The Tayside Profile for Dementia Planning is an instrument designed to obtain data for population needs assessment and planning. It provides a brief tool to collect a minimum dataset by non-specialists. Third-party informants-informal carers or involved professionals-are used as data sources. The key concept is the use of a descriptive profile rather than a summative score or categorization. The profile consists of a set of needs indicators, information on current service response and demographic and background data. Key levels of dependency are measured by time interval dependency. Validity, reliability, acceptability and usability are satisfactory, with the crucial exception that informal carers and professionals appear to perceive needs differently. Further research is needed to assess which type of informant provides the more useful data. PMID- 9215947 TI - What depressive symptoms are reported in Alzheimer's patients? AB - PURPOSE: To ascertain the nature of depression-related symptoms in AD. METHOD: The Hamilton Rating Scale for Depression (HAM-D) was administered as a semi structured interview to 30 consecutive Alzheimer's disease (AD) patients who also underwent independent psychiatric evaluation. The HAM-D was also administered with a caregiver as the informant. RESULTS: There was no relationship between the number of symptoms reported by patients or caregivers and patients' level of cognitive impairment. Symptom reports by caregivers living in the same household did not differ significantly from symptom reports by caregivers living elsewhere. Caregivers rated AD patients as having significantly more depressive symptoms than did patients themselves. The items most frequently endorsed by caregivers were psychic anxiety (77%), suspiciousness (50%), low energy (50%) and depression (43%). The items most frequently endorsed by AD patients were weight loss (36%), psychic anxiety (33%) and somatic anxiety (33%). Depression was endorsed by 20% of patients. Caregiver-respondent HAM-D scores suggested clinically significant depression in 27% of cases, but AD patients' scores suggested clinically significant depression in only 7% of cases. No case of major depression was found on psychiatric examination. CONCLUSIONS: Depressive symptoms seemed more an executive function loss than of primary mood disturbance in that guilt, suicidal rumination and self-perceived loss of interest were uncommon, suggesting that simple environmental measures might be the most appropriate treatment of these symptoms. PMID- 9215948 TI - Clock drawing test in elderly schizophrenia patients. AB - OBJECTIVE: Clock drawing has been studies in Alzheimer's disease but not in elderly schizophrenics. We examined clock drawing ability in elderly schizophrenia patients and sought possible correlations with demographic, clinical and cognitive variables. DESIGN: Retrospective analysis of the clock drawing item from the Cambridge Cognitive Examination (CAMCOG) presented to three independent raters. SETTING: Long-stay 'open' departments of a public psychiatric hospital in Israel. PATIENTS: Thirty-one physically well psychiatric inpatients suffering from schizophrenia (DSM-III-R, APA), between ages 60 and 76 years. MEASURES: The Clock Drawing Interpretation Scale (CDIS). RESULTS: The mean CDIS score was 14.4 (out of 20), and 61-84% of patients scored beneath the normal range (> 18). Interrater reliability was high (0.91-0.96). A moderate but significant correlation was found between CDIS and duration of illness as well as total scores on the Manchester Scale, the CAMCOG and the Mini-Mental State Examination, but not with the other variables studies. CONCLUSIONS: Clock drawing skills of a significant portion of long-term institutionalized elderly schizophrenics are impaired. When this test is used as a screening device for Alzheimer's disease in these patients, the results should be interpreted cautiously. Clock drawing abilities in these patients seem to be related to cognitive and non-cognitive (psychiatric state) factors, as well as to illness duration. PMID- 9215949 TI - Symptomatological characteristics distinguish between frontotemporal dementia and vascular dementia with a dominant frontal lobe syndrome. AB - OBJECTIVE: Our hypothesis was that patients with vascular dementia and a dominating frontal lobe syndrome have a symptomatology that reflects a more widespread lesion compared with patients with frontotemporal dementia. DESIGN: Patients with vascular dementia and a dominating frontal lobe syndrome (VAD-F; N = 11) were compared with regard to clinical symptoms and imaging features on CT scans of the brain with patients with frontotemporal dementia (FTD; N = 21). SETTING: A neuropsychiatric diagnostic ward. PATIENTS: Thirty-two inpatients, aged 48-78 years, with frontotemporal dementia or vascular dementia. MEASURES: Relatives were questioned about the initial symptoms. At the clinical investigation, mental and neurological symptoms and signs were recorded using the STEP method (stepwise comparative status analysis). CT scan features of the brain were evaluated by a trained neuroradiologist. The GBS-i (Gottfries-Brane-Steen, intellectual variables) scale was used to measure the degree of dementia. RESULTS: At the onset of dementia, loss of memory (p < 0.001), sudden onset (p < 0.001), confusion (p < 0.05) and unspecified neurological signs (p < 0.05) had been significantly more frequent in the VAD-F group. At the time of the clinical investigation, lack of social awareness and presence of primitive reflexes were more frequent in the FTD group (p < 0.01 and p < 0.05, respectively) and visuospatial deficits more frequent in the VAD-F group (p < 0.05). CT of the brain showed that, apart from brain infarcts (present only in the VAD-F group), paraventricular leukoaraiosis was significantly more pronounced in the VAD-F group (p < 0.05). The groups did not differ with respect to age, age at onset or level of dementia. CONCLUSION: The findings support our hypothesis. PMID- 9215950 TI - Psychiatric symptoms of dementia among elderly people with learning disabilities. AB - OBJECTIVE: To determine the rate of psychiatric symptoms among elderly people with learning disabilities who have dementia. DESIGN: Survey. SETTING: The general community of a county in the UK. PARTICIPANTS: The whole population of people with learning disabilities who were 65 years or over. The total population was 143, of whom 134 participated (93.7%). From the total population, those with dementia were determined (N = 29). MEASURES: Description of psychopathology, using a semi-structured psychiatric rating scale. RESULTS: Psychotic symptoms occurred in 27.6%, with the most common types being delusions of thefts, other persecutory delusions and visual hallucinations of strangers in the house. The onset of other psychiatric symptoms as part of the dementia was also common, in particular changed sleep pattern, loss of concentration, worry, reduced quantity of speech, change in appetite and onset of or increase in aggression. CONCLUSIONS: People with leaming disabilities are living longer, and so the number with dementia is rising. Psychiatric symptoms occur commonly in dementia, can cause significant distress and require recognition, understanding and the development of effective managements. PMID- 9215951 TI - 'Do not resuscitate': How? why? and when? AB - OBJECTIVE: The main objective was to discover who had 'Do Not Resuscitate' (DNR) status, why, how, when and by whom these decisions were made. DESIGN, SETTING AND PATIENTS: The medical and nursing notes of all inpatients (139) (age range 16-100 years) in an inner city district general hospital on a single day were examined to determine the resuscitation status, age, sex, and diagnosis of each patient. RESULT: A decision not to resuscitate had been taken in 28 (20%) of the cases. 'Do Not Resuscitate' (DNR) patients were significantly older and more likely to suffer from malignant and cardiorespiratory disease. Patients with dementia and other psychiatric disorders were not significantly more often labelled DNR. Evidence of consultation for these decisions was lacking and the recording erratic. CONCLUSIONS: (1) There is a great need to devise and implement comprehensive guidelines. (2) There is need for appropriate and comprehensive documentation outlining the reasons why and how the decision was taken, who was consulted and review date. (3) This is an important area for audit. PMID- 9215952 TI - Predictors of change and continuity in home care for dementia patients. AB - OBJECTIVE: To investigate predictors of change in the sense of competence of primary caregivers and continuity in home care for dementia patients. DESIGN: A prospective longitudinal study with a follow-up period of 10 months. SETTING: Dementia patients living in the community selected by Dutch general practitioners. SUBJECTS: Pairs of demented patients and their primary caregivers (N = 138). MAIN OUTCOME MEASURES: Sense of competence: a 27-item scale (alpha = 0.79) based on issues derived from the family crisis model and the Burden Interview. Continuity in home care is determined by the number of patient's admissions to a nursing or retirement home. RESULTS: Regression analysis revealed that a change in the caregiver's sense of competence was independently predicted by characteristics of the patient, the primary caregiver and the professional social network. A decreased sense of competence was associated with a longer duration of dementia and the patient's more agitated behaviour, the caregiver's higher initial sense of competence and being a female caregiver sharing a household with the demented patient. A positive influence on the change in the sense of competence was found when these females received a professional intervention consisting of support for the caregiver. Reporting to be a Catholic or a Protestant compared with not being religiously involved positively influenced the change in sense of competence. Logistic regression analysis identified that continuity in home care was predicted by characteristics of the demented patient and the professional social network of the patient. Predictors of continuation of home care were: lower severity of dementia, patient's higher ADL impairment, the intervention and involvement of regular home help. Institutionalization was more likely when the patient's behaviour was more apathetic and a district nurse was involved in the care. CONCLUSIONS: Caregiver characteristics influenced the change in sense of competence but did not influence the risk for institutionalization. Findings suggest that health professionals should pay attention to the negative consequences of agitated behaviour and to the most vulnerable group, females sharing a household with the demented patient. PMID- 9215953 TI - Aggressive behaviour in the Chinese elderly--validation of the Chinese version of the rating scale for aggressive behaviour in the elderly (RAGE) in hospital and nursing home settings. AB - OBJECTIVE: The objective is to examine the validity of the Chinese version of the Rating Scale for Aggressive Behaviour in the Elderly (RAGE) in Hong Kong. DESIGN: A cross-sectional study comparing the pattern of aggressive behaviour among residents of different elderly institutions. SETTING: A nursing home and a psychogeriatric inpatient unit. PATIENTS: Psychogeriatric inpatients and nursing home residents. Thirty subjects participated in the validation study of the Chinese version of the RAGE (CRAGE). Eighty-eight subjects were assessed by the CRAGE for pattern of aggressive behaviour. MEASURES: The CRAGE and the Chinese version of the Mini-Mental State Examination (CMMSE). RESULTS: The CRAGE showed satisfactory validity and reliability measures. Aggressive episodes were found in 57% of the subjects, mostly mild in nature. No significant difference was found in the CRAGE total scores in different institutions and across diagnoses. Hospital and schizophrenic patients had significantly higher ratings in overall aggressiveness. Demented subjects had higher CRAGE ratings with CMMSE scores from 11 to 15. CONCLUSIONS: CRAGE is a valid instrument for use in the Chinese elderly. Although there is no significant difference in total aggressive episodes among different elderly institutions, chronic psychiatric patients were more frequently regarded as aggressive. PMID- 9215954 TI - A roving clinic for residential homes. PMID- 9215955 TI - Determination of the binding of a beta 2-blocker drug, frusemide and ceftriaxone to serum proteins by capillary zone electrophoresis. AB - A modified Hummel-Dreyer method was used to study the binding of drugs with serum proteins by high performance capillary electrophoresis. The study was carried out to check the possible interaction between serum proteins and a highly selective beta 2-blocker, ICI 118551 (ICI). To prove the suitability of the method the protein binding of frusemide and ceftriaxone, drugs previously investigated, was also studied. The analyses were carried out by injecting a solution of s alpha(1) acidic glycoprotein (alpha(1)-AGP) or human serum albumin in 70 mM NaH2PO4/Na2HPO4(pH 7.4) buffer into an uncoated fused silica capillary filled with the same buffer. In the capillary, maintained at a working temperature of 35 degrees C, a known amount of the ICI, frusemide or ceftriaxone was added. The method allows the bound drug to be determined directly. PMID- 9215956 TI - Rapid resolution of drugs and related substances with an eCAP polyamine coated capillary. AB - The long term stability of a commercial polyamine coated capillary (eCAP) is described. The capillary, which can be used in the CZE and MEKC mode, is based on coating with a polyamine after conditioning with 1 M NaOH and regeneration of this coating after each run. The stability was tested over 6 months on the drug trimethoprim and the R.S.D. values for migration time and peak area were 2.86 and 3.62% respectively (n = 8, each time of determination) (> 600 sample injections over the period). This stability was utilised in the validated method developed for trimethoprim and four of its related impurities. The repeatability of peak area for trimethoprim (without normalisation or external standard) was, within day R.S.D. = 1.02% (n = 8) and between-days R.S.D. = 2.02% (n = 8 each day). Linearity was good (for 50 micrograms ml-1 target) (y = 249.6x + 17.3 (r = 0.992, n = 6). These results for trimethoprim and for other drug mixtures were comparison with conventional capillaries and the advantage of reducing the polyamine treated eCAP capillary to a minimum length is described, to achieve rapid assay of the 5 component timethoprim mixture in < 2 min. PMID- 9215957 TI - A chiral capillary electrophoresis method for ropivacaine hydrochloride in pharmaceutical formulations: validation and comparison with chiral liquid chromatography. AB - A capillary electrophoresis method for the determination of the enantiomeric purity of the local anaesthetic ropivacaine hydrochloride in injection solutions has been validated. The method showed the required limit of quantitation of 0.1% enantiomeric impurity. Good performances were shown for specificity, linearity, system repeatability, intermediate precision and accuracy. Robustness was tested via a full factorial design at two levels and the method proved to be robust. Comparison of the capillary electrophoresis method with the liquid chromatographic method currently used for several years at our laboratory on real samples of ropivacaine injection solutions showed that the techniques do not give significantly different results. PMID- 9215958 TI - Optimized methods for capillary electrophoresis of tetracyclines. AB - Optimized methods for the analysis of some tetracyclines by capillary electrophoresis are described. Different buffer systems were employed for the separation of tetracycline, oxytetracycline and demeclocycline from their respective major impurities, including the 2-acetyl-2-decarboxamido derivatives. The influence of buffer pH and buffer concentration was systematically investigated. Non-ionic surfactant Triton X-100 and methyl-beta-cyclodextrin were used to obtain improved selectivity in the case of oxytetracycline and demeclocycline. The results are compared with those of previously established liquid chromatography methods. Good correlations were obtained. PMID- 9215959 TI - Micellar electrokinetic chromatography stability indicating assay and content uniformity determination for a cholesterol-lowering drug product. AB - This study describes a specific, linear, precise, accurate and sensitive method for the determination of a developmental cholesterol-lowering drug formulated in capsules. The method can also determine two known hydrolytic degradants of the drug. Samples are dissolved in acetonitrile-phosphate buffer pH 4.5, diluted with water and assayed by micellar electrokinetic chromatography (MEKC) in a buffer containing 0.1 M borate-0.025 M SDS at 30 degrees C with an applied voltage of 25 kV. Detection is by UV absorbance at 200 nm. The method was cross validated by comparison with a gradient elution HPLC method. The MEKC method gave at least equivalent precision, accuracy and sensitivity to HPLC but was superior in the resolution of the known impurities and gave a considerably shorter analysis time. The method has been accepted as part of a regulatory submission to the US Food and Drug Administration (FDA). PMID- 9215960 TI - Validation of a simple method for the determination of oxytetracycline in ointment by non-aqueous capillary electrophoresis. AB - A non-aqueous capillary electrophoresis method for the determination of oxytetracycline in an ointment have been validated. The oxytetracycline (OTC) is separated from related impurities and degradation products as metal chelates with magnesium ions. Thus, the method show high selectivity. The sample preparation is performed as a single extraction step of OTC from the melted ointment. The test for linearity in the range from 0.2-3.0 mg ml-1 gave a straight line with a coefficient of correlation greater than 0.999. Precision and accuracy were investigated using standard addition at three different concentration levels and six separate determinations at each level. The precision is good and may be expressed as the coefficient of variation which was lesser than 3.6%. The recovery is close to 100%. PMID- 9215961 TI - Comparison of aqueous and non-aqueous capillary electrophoresis for quantitative determination of morphine in pharmaceuticals. AB - Two capillary electrophoresis methods involving aqueous and non-aqueous electrophoresis media, respectively, have been compared for the quantitative determination of morphine in pharmaceutical preparations. In the aqueous system the separation from other opium alkaloids was achieved using 2,6-di-O-methyl-beta cyclodextrin as an additive to the electrophoresis buffer. In the non-aqueous system no other additives than the electrolytes were necessary in order to achieve separation of the opium alkaloids. The two methods have been partially validated and compared with a currently used high-performance liquid chromatography method. From the overall point of view the validations show that the three methods are equivalent in performance and that they are appropriate for the purposes they are intended for. PMID- 9215962 TI - Validated capillary electrophoresis method for the analysis of a range of acidic drugs and excipients. AB - A capillary electrophoresis (CE) method employing a high pH borate buffer has been validated to allow analysis of a wide range of acidic compounds including active drugs, pharmaceutical formulations, excipients, starting materials and intermediates. An internal database has been established to demonstrate the wide applicability of the method. The method has been extensively validated and is in routine use in a number of our laboratories worldwide. In particular, acceptable injection precision is obtained through the use of internal standards and the method robustness was evaluated using an experimental design. The method allows a number of cost and time saving benefits. PMID- 9215963 TI - The use of capillary electrophoresis as part of a specificity testing strategy for mitoguazone dihydrochloride HPLC methods. AB - Capillary electrophoresis (CE) has been used as part of a validation experiment designed to prove the specificity of high performance liquid chromatography (HPLC) methods used for analysis of mitoguazone dihydrochloride drug substance. Data regarding accuracy, precision and sensitivity of the CE methods are presented as well as a comparison of results obtained from CE, HPLC and thin layer chromatography (TLC) analysis of samples stressed under a variety of conditions. It was concluded that, not only were the HPLC methods being investigated specific, but that CE could potentially be used to replace HPLC for the routine assay of mitoguazone dihydrochloride. PMID- 9215964 TI - Determination of six water-soluble vitamins in a pharmaceutical formulation by capillary electrophoresis. AB - A method was developed for the quantitative analysis of six water-soluble vitamins (thiamine, nicotinamide, riboflavine, pyridoxine, ascorbic acid and pantothenic acid) in a pharmaceutical formulation, using free solution capillary zone electrophoresis (CZE) in uncoated fused silica capillaries and UV detection. The influence of different parameters, such as the nature of the buffer anionic component and buffer concentration on the CZE separation of vitamins was investigated using four vitamins of the B group as model compounds. A good compromise between resolution, analysis time and analyte stability was obtained by use of a 50 mM borax buffer of pH 8.5. This CZE method was found to be very useful for the separation of more complex samples, a mixture of ten water-soluble vitamins being completely resolved in about 10 min. However, cyanocobalamine could not be separated from nicotinamide in this CZE system, the two compounds being in uncharged form at the pH used. These two compounds could easily be resolved by micellar electrokinetic chromatography (MEKC), the anionic surfactant dodecylsulfate being added to the running buffer at 25 mM concentration. In the pharmaceutical formulation, some excipients were found to be adsorbed to the capillary surface, giving rise to a progressive decrease of the electroosmotic flow and consequently to a simultaneous increase of analyte migration times. A capillary wash with sodium hydroxide had to be made between successive runs in order to minimize these effects. Good results with respect to linearity, precision and accuracy were obtained in the concentration range studied for the six vitamins, using nicotinic acid as internal standard. PMID- 9215965 TI - Qualitative and quantitative measurements of oligonucleotides in gene therapy: Part II in vivo models. AB - Part I of this review has already been presented on methods of processing and purification of crude or raw samples in cell-culture or cell free systems. Part II of the review focuses on the in vivo models of determination of input oligonucleotides in both non-primates and primates. Emphasis has been given to the techniques developed for quantification of oligonucleotides or their metabolites from biological samples including blood, plasma, serum, urine and other tissues. PMID- 9215966 TI - The effect of gamma radiation on the degradation of salbutamol. AB - The use of ionizing radiation for sterilization of pharmaceuticals is now a well established technology. Degradation of salbutamol was investigated after gamma irradiation using HPLC and ESR spectroscopy. HPLC evidenced the formation of radiolytic products after gamma irradiation. Salbutamol showed a degradation of nearly 2% at 25 kGy. Sterilization of salbutamol in the liquid state appeared not technically feasible. Simulation of the increase of free radicals versus dose was performed using linear and polynomial regression. These radicals could be detected even after a storage period of more than 12 months. PMID- 9215967 TI - Foundations of chemical microscopy. 1. Solid-state characterization of 5 nitrobarbituric acid (dilituric acid) and its complexes with group IA and group IIA cations. AB - 5-Nitrobarbituric acid (dilituric acid) has been used as a chemical microscopic reagent for the qualitative identification of alkali metal (Group IA) and alkaline earth (Group IIA) cations. This methodology was based on the characterization of observed crystal morphologies, since a unique crystal habit could be associated with each adduct product. To understand the scientific foundations which permitted chemical microscopy to function as a useful analytical technique, the products formed between dilituric acid and the Group IA and IIA cations were characterized using polarizing optical microscopy, powder X ray diffraction, thermal analysis and solid-state nuclear magnetic resonance. It was found that the origins of the different crystal morphologies associated with each of the adduct arose from the ability of the systems to form various hydrate species, which could also contain structural variations due to cation/diliturate packing patterns. PMID- 9215969 TI - Quantitation of cocaine and cocaethylene in small volumes of rat whole blood using gas chromatography-mass spectrometry. AB - A simple solid phase extraction (SPE) technique combined with gas chromatography mass spectrometry (GC/MS) operated in selected ion monitoring (SIM) mode is described for quantitation of cocaine and cocaethylene in small samples (250 microliters) of rat whole blood. Use of (N-[2H3C])-cocaine and (N-[2H3C]) cocaethylene internal standards resulted in high sensitivity and selectivity for this analytical method. Analysis was performed using a Hewlett-Packard 5890 GC equipped with a 7673A Automatic Liquid Sampler linked to a Hewlett-Packard 5972 Mass Selective Detector. Separation of analytes was accomplished on a cross linked methyl silicone gum capillary column (Ultra 1: 12m x 0.2mm (i.d.) x 0.33 microns). Linearity was established over a wide range of concentrations (5.0 2000.0 ng ml-1) with good precision. Limits of detection (LOD) were 1.0 and 2.0 ng ml-1 for cocaine and cocaethylene, respectively. This analytical method was designed for use in pharmacokinetic experiments studying the formation of cocaethylene following ethanol pretreatment in rats administered cocaine. PMID- 9215968 TI - A sensitive method for the determination of uranium in biological samples utilizing kinetic phosphorescence analysis (KPA). AB - Kinetic phosphorescence analysis is a technique that provides rapid, precise and accurate determination of uranium concentration in aqueous solutions. This technique utilizes a laser source to excite an aqueous solution of uranium, and measures the emission luminescence intensity over time to determine the luminescence decay profile. The lifetime of the luminescence decay profile and the linearity of the log luminescence intensity versus time profile are indications of the specificity of the technique for uranium determination. The luminescence intensity at the onset of decay (the initial luminescence intensity), which is the luminescence intensity at time zero after termination of the laser pulse used for excitation, is proportional to the uranium concentration in the sample. Calibration standards of known uranium concentrations are used to construct the calibration curve between the initial luminescence intensity and uranium concentration. This calibration curve is used to determine the uranium concentration of unknown samples from their initial luminescence intensity. We developed the sample preparation method that allows the determination of uranium concentrations in urine, plasma, kidney, liver, bone spleen and soft tissue samples. Tissue samples are subjected to dry-ashing in a muffle furnace at 600 degrees C and wet-ashing with concentrated nitric acid and hydrogen peroxide twice to destroy the organic component in the sample that may interfere with uranium determination by KPA. Samples are then solubilized in 0.82 M nitric acid prior to analysis by KPA. The assay calibration curves are linear and cover the range of uranium concentrations between 0.05 micrograms l-1 and 1000 micrograms l 1 (0.05-1000 ppb). The developed sample preparation procedures coupled with the KPA technique provide a specific, sensitive, precise and accurate method for the determination of uranium concentration in tissue samples. This method was used to quantify uranium in different tissue samples obtained over a period of 90 days following a single intraperitoneal uranium dose of 0.1 mg kg-1 in rats. PMID- 9215970 TI - Estimation of anthelmintic compound 81/470 in cow's milk by high-performance liquid chromatography: method development and validation. AB - CDRI compound 81/470 is a new broad spectrum anthelmintic agent and is being developed for veterinary use. HPLC assay method for 81/470 in cow milk was developed and validated. The sample preparation consisted of protein precipitation, followed by extraction with ether. Separation was achieved on a C18 column using acetonitrile-buffer (pH 6, 50 mM) mobile phase and the compound was quantitated using fluorescence detector. The recovery of 81/470 was above 90% and was consistent over the calibration range of 10-1000 ng ml-1. Accuracy and precision were determined by analyzing replicate samples and were found to be within acceptable limits. Five freeze-thaw cycles of spiked milk and storage of processed dry residues at -30 degrees C for 7 days did not have any detrimental effect on the stability of 81/470. PMID- 9215971 TI - Determination of captopril in human serum by high performance liquid chromatography using solid-phase extraction. AB - A rapid, simple and sensitive assay was developed for determination of captopril in human serum. We employed silica gel cartridge for efficient extraction of captopril adduct from human serum. Captopril was trapped with p-bromophenacyl bromide (pBPB) to give captopril-pBPB adduct. A 4-ml benzene extract of 1 ml acidified serum was passed through 1 ml silica gel cartridge. Potential interfering compounds were removed with 4-ml benzene wash. The captopril-pBPB adduct was eluted with 0.5 ml acetonitrile. Of this acetonitrile solution (100 microliters) was injected on an ODS reverse phase HPLC column (chromatography conditions; mobile phase; acetonitrile-water-acetic acid (225:270:5, v/v/v), flow rate; 1 ml min-1, detection; UV at 263 nm). It is found that this method is accurate and does not require time consuming evaporation-concentration steps. Recovery exceeds 94% and analytical responses are linear over captopril concentration range from 50 up to 1000 ng ml-1. The coefficients of variation from 108 ng ml-1 to 605 ng ml-1 varied between 3.7-7.7% and the relative error did not exceed 3.7%. Therefore, this method can be used for routine clinical monitoring and in pharmacokinetic studies of captopril. PMID- 9215972 TI - HPLC analysis for amobarbital N-glycosides in urine. AB - A study was undertaken to determine if humans excrete both amobarbital N glucuronides and N-glucosides in urine after an oral dose of amobarbital. Amobarbital N-glucuronides were synthesized and characterized. A reverse phase LC method using post-column pH adjustment and UV detection at 240 nm was developed and used for the quantification of the amobarbital N-glucosides and N glucuronides in human urine. Amobarbital was administered orally to seven male subjects and the total urine was collected for a period of 48-53 h after dosing. After filtration, the urine was injected directly onto the HPLC column to analyze for the presence of metabolites. The previously identified (5S)-amobarbital N glucoside was detected in all seven subjects. The (5R)-amobarbital N-glucoside was detected at lower concentrations in only four of the subjects. At the levels at which amobarbital N-glucosides were detected, there was no evidence for the formation and excretion of the amobarbital N-glucuronides. Amobarbital N glucuronidation is not a quantitatively significant pathway for the biodisposition of amobarbital in humans. PMID- 9215973 TI - Simultaneous determination of amoxycillin and clavulanic acid in pharmaceutical products by HPLC with beta-cyclodextrin stationary phase. AB - A simple, rapid and accurate method for simultaneous determination of amoxycillin and clavulanic acid using HPLC with beta-cyclodextrin stationary phase was developed. It involves the use of tetraethylammonium acetate (TEAA) as an additive reagent, methanol-buffer solution (pH 4.5) (35:65; v/v) as the mobile phase, detection at 225 mm and chromatogram within 12 min. Linearity and precision of the internal standard method have been obtained. Recoveries ranged from 99.25 to 105.63% for amoxycillin in the synthetic mixture. For clavulanic acid it was from 99.50 to 101.64%. This method is convenient and reproducible for analyses of these two components in different dosage forms. PMID- 9215974 TI - On randomized clinical trials. PMID- 9215975 TI - Use of the C-scan in evaluation of peripheral lymphedema. AB - The C-(cutaneous)-scan constitutes a simple way to evaluate the status of the lymphatic system of swollen and apparently normal extremities. We have termed the procedure C-scan because this expression emphasizes the injection of the radiotracer into the cutis. Only one scintiscan, which takes approximately 15 minutes, is performed 3 hours after injection, in contrast to other investigative procedures including standard lymphangioscintigraphy, which utilize multiple scannings that unnecessarily prolong the duration of the study. The C-scan method is semi-quantitative and distinguishes pathologic conditions of the lymphatic system by means of whole body scintigraphy and by measuring the radioactivity in specific regions of interest rather than by examining solely differences in delay of transport. Both a qualitative image and a quantitative assessment are thereby produced. The C-scan is performed using a general purpose gamma camera in line to a multiprocessor computer system. Scinti-scanning is performed 3 hours after an intracutaneous injection of 20 MBq of Tc99m nanocolloid (Solco) distally into the dorsum of the extremity. The image obtained is divided into regions of interest and the radioactivity in these regions is calculated as the percentage of the total radioactivity counted. The C-scan is easy to perform and can be repeated if necessary every 2 days. The effects of various treatments of lymphedema, either nonoperative or operative, can be assessed short-term, and examples are provided. In conjunction with limb volume measurements, the C-scan gives an accurate impression of changes in the lymphedematous extremity before, during, and after treatment. Applications of this imaging technique are shown after combined "decongestive" physiotherapy, lymph-venous anastomosis, ablative (debulking) surgery, liposuction, and thermotherapy. PMID- 9215976 TI - Expression of interleukin-6 receptors and NF-kappa B in AIDS-related Kaposi sarcoma cell strains. AB - AIDS-related Kaposi sarcoma (AIDS-KS) is the most common malignancy associated with HIV infection, with an incidence of 10-30% of all AIDS patients. As such, there have been a large number of AIDS-KS cell strains isolated and numerous studies conducted to elucidate the mechanisms of malignancy in this disease. We have reported histological grade associated differences in the ability of AIDS-KS cell strains to proliferate under conditions of minimal growth factor supplementation, with strains derived from high grade lesions having enhanced proliferation potential. Furthermore, we found that this difference in in vitro growth characteristics was not attributed to grade associated differences in autologous growth factor release. These current investigations explored the hypothesis that grade associated growth differences could be attributed to differences in the expression of the components of the IL-6 receptor, or expression/inducibility of the pleotrophic transcription factor NF-kappa B. We determined there were no significant grade associated differences in the expression of either component (IL-6R alpha chain or gpl30) of the IL-6 receptor. However, non-lesional oral derived cell strain lysates from AIDS-KS patients (n = 4) contained significantly lower concentrations of both components of the IL-6 receptor than AIDS-KS strains (n = 8) and lower concentrations of gp-130 than normal human oral derived fibroblasts (n = 2). Comparative analysis of sera concentrations of soluble components of the IL-6 receptor did not demonstrate significant differences between HIV+/KS+ (n = 7), HIV+/KS- (n = 9) and normal (HIV-/KS-) (n = 4) populations. Further, no differences were detected in the expression of NF-kappa B in AIDS-KS cell strains (n = 5) derived from both high and low histological grade lesions as compared to nonlesional AIDS-KS cell stain (n = 1) and normal human oral derived fibroblasts (n = 2) under conditions of: constitutive/proliferative growth, sera starvation, oxidative stress, and mitogen reintroduction after sera starvation. In conclusion, these investigations have eliminated two explanations for histological grade associated differences for in vitro growth potential of AIDS-related KS cell strains and further substantiated the lack of systemic paracrine cytokine/cytokine receptor effects in AIDS-KS pathogenesis. PMID- 9215978 TI - Lymphatic dysfunction in conjunction with dysregulated hyperdynamic blood flow (the hyperstomy syndrome). PMID- 9215977 TI - Validation of an optoelectronic limb volumeter (Perometer). AB - The Perometer, a device designed for the measurement of limb volume, has been rigorously assessed by comparison with other methods. Differences in the volume of geometric shapes and limbs determined by the Perometer and a tape measure/meter rule (i.e., Perometer minus direct measurement) were -0.8 to 2.4% (cylinders), -4.6% (truncated cone), -3.3% (mannequin limbs), 6.1% (normal human arms) and 6.8% (lymphedema arms). The larger differences were likely to be due to deviation from circular or elliptical cross-section (Perometer or tape method) and compression of the arm (tape method). Errors arising from incorrect positioning within the measuring frame were generally small, but large errors occurred when a cylinder was partially rotated within the frame (i.e., no longer perpendicular to the light beams). The Perometer was highly reproducible, each measurement taking only a few seconds. When recording the change in volume with time of a segment of arm during venous occlusion (blood flow measurement by venous occlusion plethysmography) using the Perometer plus a mercury strain gauge, between-method differences for individual blood flow recordings were apparent. The source of these differences is discussed. However, using the average of a number of blood flow recordings the Perometer and the strain gauge agreed fairly closely for both the normal and lymphedema arms. The Perometer is thus a reliable and convenient tool for the measurement of limb volume, and may also be used to measure the rate of swelling during venous occlusion plethysmography. PMID- 9215979 TI - [Investigation and action concerning health inequalities]. PMID- 9215980 TI - The relevance of axonal swellings and atrophy to gamma-diketone neurotoxicity: a forum position paper. AB - Traditionally, gamma-diketone neuropathy is classified as a distal axonopathy and has been characterized by giant axonal swellings in CNS and PNS tissues. These swellings contain neurofilamentous masses and are associated with thinning and retraction of the myelin sheath. It has been proposed that this axonopathy is caused by direct gamma-diketone modification of neurofilaments (NFs) involving pyrrolation of epsilon-amino groups on NF lysyl residues and possibly secondary autoxidation of the pyrrole rings with creation of covalent NF-NF crosslinks. Neurofilaments are thought to undergo chemical modification as they progress along the axonal axis and, eventually, accumulate at distal nodes of Ranvier where their proximodistal movement is impeded. Development of swelling presumably initiates axonal degeneration and subsequent functional deficits. However, other research suggests that axonal swellings are a non-specific effect related to subchronic gamma-diketone exposure. Such evidence draws into question the mechanistic relevance of these swellings. In contrast, research conducted over the past decade indicates axonal atrophy is a specific morphologic component of gamma-diketone neuropathy which might have both functional and mechanistic importance. In this overview, the potential neurotoxicological significance of both axonal swellings and atrophy are evaluated critically. Based on the evidence to be presented, we propose that axonal atrophy is the morphological consequence of the molecular mechanism of gamma-diketone neuropathy. Accordingly, several mechanistic scenarios related to the development of atrophy will be discussed. It is hoped that this Forum will stimulate scientific debate and initiate laboratory investigations which will either confirm or refute the involvement of axonal atrophy in gamma-diketone neurotoxicity. Investigating gamma-diketone atrophy might provide insight into the mechanism of other toxic axonopathies which are also associated with reduced axon caliber; e.g., acrylamide and carbon disulfide neuropathies. PMID- 9215981 TI - Specificity of neurofilament swellings and axonal atrophy: commentary on forum position paper. PMID- 9215983 TI - Protein crosslinking correlates with axonal atrophy, swelling and degeneration in gamma-diketone neuropathy: commentary on forum position paper. PMID- 9215982 TI - Axonal swellings vs atrophy: what is the relevance to gamma-diketone peripheral neuropathy? Commentary on forum position paper. PMID- 9215984 TI - Mechanistic importance of axonal atrophy and swelling in gamma-diketone axonopathy: commentary on forum position paper. PMID- 9215985 TI - Neurofilamentous swellings and axonal atrophy are of unclear significance to diketone toxicity: commentary on forum position paper. PMID- 9215986 TI - Neurotoxicology: from Cinderella to Cordelia's secret. PMID- 9215987 TI - Pb2+ inhibits NMDA receptor function at high and low affinity sites: developmental and regional brain expression. AB - Lead (Pb2+) is a potent inhibitor of the NMDA receptor complex. In the present study we have measured high and low affinity components of NMDA receptor inhibition by Pb2+ using [3H]-MI-801 binding as a biochemical indicator of NMDA receptor function. The inhibitory effects of Pb2+ were dependent on the age of the rat and the brain region analyzed. The number of [3H]-MK-801 binding sites associated with the high and low affinity sites of Pb2+ inhibition in the hippocampus increased as a function of age, peaking at postnatal day 28 and 21, respectively. In the cerebellum, a steady decrease in the number of both types of Pb(2+)-sensitive [3H]-MK-801 binding sites was measured from postnatal day 1 to adulthood. High and low affinity Pb(2+)-sensitive [3H]-MK-801 binding sites were measured in the cerebral cortex in early development, but the resolution of the two sites was not possible after postnatal day 14. The Pb2+ sensitivity of the NMDA receptor complex also appeared to be regulated developmentally. PMID- 9215988 TI - Structural neurotoxicologic investigation of the glycine antagonist 5-nitro-6,7 dichloroquinoxalinedione (ACEA-1021). AB - NMDA antagonists of glutamate have psychotomimetic side effects and structural side effects which have been shown to be lethal to CNS neurons in the cingulate and retrosplenial cortex of rodents, yet these compounds may reduce focal ischemic brain damage. This investigation used 38 Wistar rats to determine whether the structural toxicologic profile of a newly developed halogenated quinoxalinedione derivative, a pharmacologic antagonist of the glycine site on the NMDA receptor complex, is identical to that seen with MK-801. In the cingulate and retrosplenial cortex, examination of glutaraldehyde perfusion fixed, plastic-embedded tissue 4 to 6 hours after intravenous administration of 10 mg/kg of the glycine antagonist 5-nitro-6,7-dichloroquinoxalinedione (ACEA 1021), no changes were seen by light or electron microscopy. At a dose of 30 mg/kg, neurons were seen containing 1 to 2 microns granules in perikarya and axons. Vacuolated neurons, as described in NMDA-antagonist neurotoxicity, were exceedingly rare, comprising only 4 in the entire study. Electron microscopy of the granulated profiles showed intracytoplasmic areas containing grouped mitochondria and lysosomes, located in neuronal perikarya, and rarely in myelinated axons. Neuronal necrosis was evaluated in formaldehyde perfusion fixed, paraffin-embedded tissue at one week survival, and was absent. MK-801 5 mg/kg, in contrast, caused irreversible (necrotizing) neuronal changes. The results demonstrate that this glycine antagonist is devoid of lethal neurotoxicity, but causes a reversible alteration in a small proportion of cingulate and retrosplenial cortical neurons. Since previous studies have shown anti-ischemic efficacy of this compound in focal, but not global ischemia, it appears that the therapeutic profile of this antagonist of the strychnine insensitive glycine site is similar, but the toxicologic structural profile is different, from NMDA receptor antagonists. PMID- 9215989 TI - Tau in aluminum-induced neurofibrillary tangles. AB - Aluminum is a neurotoxin and in susceptible species induces a neurofibrillary pathology characterized by argentophilic masses in neuronal perikarya and in axonal spheroids. These inclusions are known to contain neurofilament proteins. Using immunocytochemistry and immunoblotting, we demonstrate that tau is a component of these aluminum-induced neurofibrillary tangles (Al-NFTs) in rabbits. Double-label immunocytochemistry experiments reveal co-localization of phosphorylated neurofilaments (using SMI31) and tau (using tau-1, tau-5, AT8, and PHF-1) in the perikaryal Al-NFTs. Non-phosphorylated tau (detected using tau-1) occupies a smaller area of the Al-NFT than the total pool of tau proteins (detected using tau-5). The area of total tau and non-phosphorylated tau immunolabeling in the Al-NFT increases as the size of the Al-NFT (i.e., the proportion of cell area occupied by the Al-NFT) increases. The proportion of cell area (outside of the Al-NFT) occupied by tau (as indicated by tau-5) decreases as the area of tau in the Al-NFT increases and as the size of the Al-NFT in the cell increases. Immunoblotting experiments demonstrate 1) the specificity of the tau antibody labeling and verify a lack of cross-reactivity of the tau-5 antibody to neurofilament proteins in rabbit tissue; and 2) no alterations in the levels of tau resulting from aluminum-treatment. These data suggest that as the size of the Al-NFT in a cell increases there is less tau in the neuronal perikarya. Therefore, there may be less tau in the perikarya available to perform normal functions such as microtubule polymerization and stabilization. Tau and neurofilament proteins are perturbed in a number of neurodegenerative disorders such as Alzheimer's disease, diffuse Lewy body disease, and Parkinson's disease. Aluminum-induced neurofibrillary pathology may provide a model to study perturbation in tau and neurofilaments, their phosphorylation and deposition into pathological inclusions. PMID- 9215991 TI - Histological, histochemical and autoradiographic evidence of in vitro neurotoxic effects of the novel antitumor agent, 9-methoxy-N2-methylellipticinium acetate. AB - 9-Methoxy-N2-methylellipticinium acetate (MMEA) exhibits selective cytotoxicity towards glial-derived human brain tumor cell lines comprising the U.S. National Cancer Institute preclinical drug screen. Neurotoxic potential of MMEA has been demonstrated in an in vitro model employing sagittal slices of rat brain. Histochemical staining of rat brain slices for lactate dehydrogenase (LDH) activity revealed decreased staining intensity following incubation with increasing concentrations of MMEA (0.1-100 microM). Cytological evaluation of paraffin sections stained with Cresyl Fast Violet revealed neuronal damage delineated by cytoplasmic vacuolation, and distention and fraying of the plasma membrane. No glial or vascular pathology could be discerned. Autoradiography, following exposure to 14C-MMEA, revealed distinct labelling of the large neurons of the brain stem, neurons in the thalamus and pyramidal neurons of the hippocampus, indicating neuronal uptake of the drug. PMID- 9215990 TI - Neuronal gene expression in aluminum-induced neurofibrillary pathology: an in situ hybridization study. AB - Alterations in cytoskeletal proteins such as the perikaryal accumulation of neurofilaments (NFs) occur in a number of human neurodegenerative diseases including Alzheimer's disease, Parkinson's disease, and amyotrophic lateral sclerosis and may contribute to their debilitating effects. The administration of aluminum salts to rabbits induces the aberrant accumulation of NFs within the proximal axons and perikarya of vulnerable neurons and is one animal model which has been extensively studied in an attempt to gain insight into the mechanism(s) of NF perturbations in human disease. Previous studies using Northern blotting techniques to examine mRNA levels in the aluminum-induced neuropathy model have led to seemingly contradictory results. We have used in situ hybridization which provides the cellular resolution needed to: 1) determine whether there are generalized decreases in the levels of mRNA expression or decreases in mRNA encoding specific proteins; 2) determine whether alterations in mRNA levels occur specifically in neurons with NF accumulations; and 3) begin to resolve some of the apparent contradictions in the literature. A moderate dose of aluminum lactate administered on two consecutive days produced neurofibrillary tangles in spinal cord neurons seven days after the first dose. Polyadenylated mRNA levels were not altered in spinal cord neurons in aluminum-treated compared to saline treated control animals or in tangle-bearing compared to non tangle-bearing neurons in aluminum-treated animals. Middle and high NF subunit (NFH) mRNA levels were not significantly different from polyadenylated mRNA levels in spinal cord neurons in aluminum-treated/control animals. NFH mRNA levels were decreased in neurons containing aluminum-induced NF accumulations. These results suggest that NFH gene expression may be down regulated by an inhibitory feedback mechanism induced by perikaryal accumulations of NFs. This inhibitory feedback regulation for NFH may have implications for neurodegenerative diseases in which NFs accumulate in neuronal perikarya. PMID- 9215992 TI - In vitro neurotoxicity of the antitumor agent 9-methoxy-N2-methylellipticinium acetate (MMEA): role of brain cytochrome P-450. AB - 9-methoxy-N2-methylellipticinium acetate (MMEA) is representative of a series of quaternized ellipticine derivatives that are selectively cytotoxic to human brain tumor cell lines derived from non-neuronal (glial) cells (Acton et al, 1994). In an attempt to determine whether MMEA may exhibit toxicity to normal brain cells, we have examined the effect of the drug, in vitro, using sagittal slices of rat brain. Incubation of rat brain slices in an artificial cerebrospinal fluid medium containing MMEA resulted in dose-dependent leakage of lactate dehydrogenase (LDH) into the surrounding medium. However, other subcellular marker enzymes such as Na(+)-K+ATPase (plasma membrane), cytochrome c oxidase, isocitrate dehydrogenase, NADH-dehydrogenase (mitochondrial), N-acetylglucosaminidase, acid phosphate (lysosomal), glyceraldehyde-3-phosphate dehydrogenase and enolase (glycolytic enzymes) were unaffected even at the highest tested concentrations of MMEA (10 and 100 microM). Preincubation of slices with reserpine (1 nM) or, dopamine or serotonin-specific reuptake inhibitors abolished MMEA-induced toxicity in brain slices. Pretreatment of slices with piperonyl butoxide and metyrapone, inhibitor of cytochrome P-450, also prevented the toxicity of MMEA. Further, brain slices prepared from phenobarbital-treated rats showed enhanced sensitivity to MMEA; significant leakage of LDH was observed at MMEA concentrations as low as 1 nM. The present studies demonstrate the toxicity of MMEA in rat brain slices, in vitro, and suggest a role for brain cytochrome P-450 in the neurotoxicity of MMEA [corrected]. PMID- 9215993 TI - Effects of the paralytic agent 2,4-dithiobiuret on viability and morphology of rat pheochromocytoma (PC12) cells. AB - In previous studies, 2,4-dithiobiuret (DTB) caused a delayed onset neuromuscular weakness in rats which was associated with decreased quantal content, alterations in postsynaptic ion channel properties, and abnormalities in nerve terminal ultrastructure. The latter include features typical of degenerating or diseased motor endplates as well as a marked proliferation of smooth endoplasmic reticulum (SER), swelling of mitochondria and evidence for a decreased in intraterminal calcium concentrations at early stages of intoxication (Jones, 1989, Acta Neuropathol. 78:72). These in vivo studies do not allow us to distinguish between the initial effects of DTB on the nerve terminal and those evolving as a result of disuse or secondary to its action on the muscle fiber or Schwann cells. To begin to distinguish between primary and secondary effects of DTB, we examined DTB-treated rat PC12 cells for comparable changes. The direct effects of DTB on PC12 cells included signs of general toxicity. Cell death in sparsely- plated cultures increased from 8-9% in controls to 13.7% at 10 microM for 24 hr exposure, and continued to increase in a concentration-dependent fashion to 25% mortality at 25 microM. However, between 25 and 100 microM there was little additional increase in mortality. 10 to 40 microM DTB slightly decreased the ability of both differentiated and undifferentiated cells to adhere to a substrate. This effect was independent of cell mortality. In moderately-differ entiated cells having processes up to 10 cell diameters and several varicosities, concentrations of DTB as high as those invoking increased cell mortality and comparable to those affecting the rat neuromuscular junction did not cause abnormalities in the structure of the SER. No masses of tubulovesicular profiles were seen with transmission electron microscopy, and large changes in the quantity or distribution were not detected at the light microscope with the fluorescent stains DiOC6 or rhodamine B. Other signs of neuronal degeneration (blebbing of the plasmalemma, large intracellular droplets, mitochondrial abnormalities) preceded or accompanied any evidence for abnormalities in the SER. Thus the effect of DTB on the SER at the rat motor nerve terminal may occur secondary to a more general toxic action on other cell types, or may be dependent on a level of neuronal activity not achieved in sparsely- plated cultures, or may require a greater degree of differentiation of the neuronal cells than provided by the PC12 cell model used in this study. PMID- 9215994 TI - Olfactory mucosal lesions following subcutaneous diethyldithiocarbamate (DDTC). AB - We found that a single 600 mg/kg subcutaneous dose of the chelating agent diethyldithiocarbamate (DDTC) in rats caused severe damage of the olfactory epithelium. Damage was characterized by degeneration of the receptor cells but sparing of basal cells. This degeneration was characterized centrally (in the olfactory bulb) by 50% shrinkage of glomeruli. Reactive gliosis, as judged by immunoreactivity for glial fibrillary acidic protein, was prominent in the glomeruli at one week. Glomeruli areas had recovered to control values and gliosis in glomeruli had decreased by five weeks after injection. This recovery corresponds to sparing of the regenerative cell of the olfactory epithelium. We hypothesized that DDTC may act by disrupting xenobiotic metabolic pathways requiring divalent cations. PMID- 9215995 TI - Neurotoxicity of tunicamycin on cultured cerebellar granule cells. AB - Tunicamycin (TM) is known to cause severe neurological diseases in the central nervous system of animals. Cultured cerebellar granule cells were used to investigate the direct cytotoxicity of this agent on cerebellar neurons. Results indicate that TM is a potent inhibitor of glycosylation and protein synthesis. This agent killed granule cells in a dose- and time-dependent manner. TM treatment of these neurons lead to atrophic cell bodies and condensed nuclei. This TM-induced cell death could be prevented by cycloheximide and aurintricarboxylic acid. In contrast, TM had no apparent toxicity toward nerve growth factor-differentiated PC12 cells under similar conditions. These results suggest that TM kills cerebellar granule cells in a manner similar to programmed cell death. PMID- 9215996 TI - Effect on the peripheral nervous system of the short-term intravenous administration of paclitaxel in the rat. AB - The effectiveness of paclitaxel (Taxol) in the treatment of different tumors is well-known but, on the other hand, there is little information regarding its neurotoxicity and the mechanism(s) underlying this potentially severe side effect. In this study, using behavioral, neurophysiological, morphological and morphometric methods, we evaluated the effect of intravenous administration of paclitaxel on the rat nervous system. After 2 pilot studies, 40 female Wistar rats were treated with intravenous paclitaxel via a catheter placed in the jugular vein, while 20 animals were used as controls. Paclitaxel dissolved in ethanol/Tween 80/saline (5/5/90%) was administered 5 times over a period of 10 days. At the end of the experiment half the surviving animals in each group were evaluated and sacrificed (day 11), while the rest of the rats were evaluated and sacrificed on day 25. On day 11 the treated animals had significant impairment in pain perception (tail-flick test), coordination (rota-rod test) and nerve conduction velocity in the tail nerve. At the light microscope minimal axonal damage and Schwann cell activation were observed in the sciatic nerve. At the electron microscope microtubular accumulation was present within the axon in dorsal and ventral spinal roots and in the sciatic nerve. On day 25 the behavioral tests were normal in treated rats, while the nerve conduction velocity was still moderately reduced in comparison with the controls. At the electron microscope a morphological examination evidenced that microtubular accumulation was less severe, but still evident, especially in the sciatic nerve. Morphometric determinations performed on days 11 and 25 did not evidence differences between paclitaxel-treated rats and controls. The results of this study, the first in which an extended examination of the nervous system of animals treated intravenously with paclitaxel has been carried out, suggest that short-term administration of the drug induces mainly reversible changes in the peripheral nerves and spinal roots. Microtubules seem to be the main target of paclitaxel neurotoxicity, in much the same way as has been described for its antineoplastic activity. Finally, no pathological changes were seen in the neuronal bodies of the spinal cord and dorsal root ganglia. This model may be used for further studies with combination treatments with other antineoplastic or neuroprotective agents. PMID- 9215997 TI - Trimethyltin exposure in the rat induces delayed changes in brain-derived neurotrophic factor, fos and heat shock protein 70. AB - Trimethyltin chloride (TMT) treatment in adult rats leads to limbic brain lesions that are detectable with classical neuropathological techniques 3 days after exposure. In particular, the hippocampal cells of the CA3c region are affected. The temporal and regional characteristics of TMT toxicity as reflected in changes of activity-dependent factors were studied in adult male Sprague-Dawley rats using quantitative in situ hybridization and immunohistochemistry. No significant alterations in the BDNF mRNA were detected in hippocampus and cerebral cortex 1 and 4 h after 8 mg TMT/kg. Three days after TMT, a significant increase in BDNF mRNA was detected in CA1, and increases in BDNF mRNA were also seen in cortical layers. An increase in BDNF hybridization signal was seen over scattered neurons within and outside CA3c at 3 days. Four h after 8 mg TMT/kg, BDNF immunoreactivity was reduced in the pyramidal cells of the CA3c and CA1 regions as well as in the dentate gyrus. No significant change in BDNF immunoreactivity was seen in hippocampus or cerebral cortex 3 days after TMT. BDNF interacts with the high-affinity receptor tyrosine kinase B (trkB). No immediate alteration in trkB mRNA was seen in hippocampus or cerebral cortex after 8 mg TMT/kg, while at 3 days trkB mRNA was significantly reduced in the CA3c pyramidal cell layer. No changes could be detected in neurotrophin-3 mRNA at either 1, 4 h or 3 days after TMT. Three days after 8 mg TMT/kg, a major induction of hsp70 mRNA occurred in a subset of neurons in the CA3c region, concomitant with an increased expression of c-fos mRNA as well as Fos protein in the hilar region of hippocampus. Hence, an early and transient decrease in BDNF appears to occur after TMT exposure, which is succeeded at 3 days by increases in BDNF, c-fos and hsp 70 mRNAs, concomitant with a decrease in trkB mRNA in regions known to be vulnerable to TMT. These results demonstrate that TMT causes a delayed, spatially restricted increase in activity-dependent gene expression, making TMT-induced disturbances an interesting model of neurodegenerative events. PMID- 9215998 TI - Synaptophysin immunoreactive axonal swelling in p-bromophenylacetylurea-induced neuropathy. AB - A single intraperitoneal dose of 300 mg/kg of p-bromophenylacetylurea (BPAU) induced progressive distal neuropathy in rats, prominently involving peripheral nerves and long central nervous system myelinated tracts such as the fasciculus gracilis and spinocerebellar pathways. Clinical signs assessed using a Functional Observational Battery (FOB) and in-cage observation included weakness and deficits in motor and sensory integration, definitively noted on post-dosing day 7. The signs were more pronounced upon repetition of the FOB on post-dosing day 12. The neuropathological substrate of these signs was a progressive axonopathy with regional swelling, leading to Wallerian-like degeneration of affected myelinated fibers. Immunocytochemical staining for synaptophysin revealed often striking increase in immunoreactivity for this synaptic vesicle glycoprotein in swollen and otherwise injured axons. Such accumulations were considered consistent with interruption of anterograde (and possibly retrograde) fast axonal transport systems secondary to toxicant-induced nerve fiber breakdown. PMID- 9215999 TI - L-2-chloropropionic acid inhibits glutamate and aspartate release from rat cerebellar slices but does not activate cerebellar NMDA receptors: implications for L-2-chloropropionic acid-induced neurotoxicity. AB - L-2-Chloropropionic acid (L-CPA), when orally administered at single high dose to rats produces a selective lesion in the cerebellum involving destruction of a high proportion of granule cells by a mechanism which involves N-methyl-D aspartate (NMDA) receptors. Receptor binding studies demonstrated that L-CPA a had low affinity at the glutamate and glycine binding sites at NMDA receptors (530-660 microM), respectively, whereas L-CPA did not displace [3H]AMPA, [3H]NBQX or [3H]kainate from AMPA or kainate receptors. Whole cell-patch clamp experiments using cultured granule cells failed to demonstrate changes in membrane potential of cultured granule cells when either L-CPA (0.25 or 1 microM) was added alone to the bathing solution, or in combination with glycine (10 microM). Furthermore L CPA did not alter the magnitude of the inward current produced by application of NMDA (100 microM)) to cultured granule cells, in the presence of glycine, as measured by patch clamp techniques. Experiments were also performed to discover whether L-CPA may alter the release of the excitatory amino acids from the cerebellum, which may then indirectly alter activity at glutamate receptors, leading to neuronal cell death. L-CPA (2 mM) did not affect either basal or stimulated (electrical or high potassium) endogenous aspartate release from superfused cerebellar slices nor did it alter the basal or stimulated release of [3H]aspartate from preloaded slices when introduced into the superfusion medium over 30 min. However, when cerebellar slices were preincubated with 2 mM L-CPA for 2 h at concentrations that are known to be neurotoxic to the brain in vivo, but not in vitro, the stimulated endogenous glutamate and aspartate net release was significantly attenuated, as compared to controls. Basal release was not significantly affected by the introduction of L-CPA-induced cerebellar neurotoxicity may be related to the inhibition of excitatory amino acid release from the cerebellum. In conclusion, although L-CPA does not appear to directly alter NMDA receptor activity the L-CPA-induced cerebellar neurotoxicity may be related to the inhibition of excitatory amino acid release from the cerebellum. PMID- 9216000 TI - Ethanol inhibition of reduced frequency-dependent rundown of calcium currents in acutely dissociated MS/nDB neurons from chronic in vivo lead-exposed adult rats. AB - Although it is well known that lead (Pb2+0 acutely blocks voltage-gated calcium currents (VGCCs) in mammalian neurons, little is known about the long-term effects of continuous exposure to this metal on VGCCs. In the present study, the effects of chronic lead exposure on VGCCs (with barium ions as the charge carrier) were studied using whole-cell patch-clamp electrophysiological techniques in acutely dissociated medial septum (MS)/nucleus diagonal band (nDB) neurons. Neither peak, end current amplitudes, nor the current-voltage relationship were affected by chronic lead exposure. However, VGCCs repetitively evoked at frequent 6 s intervals displayed diminished whole-cell current rundown after 2 min of stimulation in cells from chronic Pb-exposed rats compared to cells from control Na-exposed rats. Because rundown after repetitive stimulation at a slower rate (20 s intervals) was not different between Pb-exposed and Na exposed, reduced rundown at 6 s intervals was probably due to decreased slow inactivation of voltage-gated calcium channels. Interestingly, acute application of 60 mM ethanol reversed the reduced rundown in cells from Pb-exposed rats while having no effect on cells from Na-exposed rats. Clearly, acute ethanol treatment antagonized the effect of chronic lead exposure, unlike the additive interaction we observed previously with synaptic plasticity (Grover and Frye, 1996). Acute application of 1 microM Pb2+ completely blocked VGCCs similarly in neurons from Na-exposed and Pb-exposed rats. These findings do not suggest that major adaptive changes in VGCCs have occurred during chronic in vivo exposure to lead. But, subtle changes in channel efficiency only revealed under conditions of repetitive stimulation may exist, and are reversed by ethanol. These subtle changes may be sufficient to influence neuroplasticity such as LTP. PMID- 9216002 TI - Reduction of choline acetyltransferase mRNA in the septum of developing rat exposed to inorganic lead. AB - We previously reported that perinatal exposure to low levels of lead (Pb) results in a pronounced reduction of choline acetyltransferase (ChAT) activity in the septohippocampal pathway of the rat. In the present study we employed a ChAT specific 43-base oligonucleotide hybridization probe to compare the developmental expression of ChAT mRNA and ChAT activity in the septum of normal and Pb-exposed rats. Rats were exposed to Pb via maternal administration of 0.2% solution of Pb acetate in drinking water from one week before birth through postnatal day 21 (PN21). In normal animals, the developmental changes in the ChAT mRNA and activity levels were temporally correlated and approached adult levels by PN21. In the Pb-exposed pups, ChAT mRNA and activity levels were reduced respectively by 69% and 50% at PN7, 47% and 33% at PN14, and 47% and 22% at PN21. In contrast, Pb exposure had no significant effect on either ChAT mRNA or ChAT activity in the dams. These results indicate that cholinotoxicity of Pb in the septohippocampal pathway of the rat involves developmental stage-dependent disruption of ChAT gene expression. PMID- 9216001 TI - 2-Iminothiazolidine-4-carboxylic acid produces hippocampal CA1 lesions independent of seizure excitation and glutamate receptor activation. AB - We previously demonstrated that 2-iminothiazolidine-4-carboxylic acid (2-ICA), formed by cyanide reacting with cysteine, caused glutamate antagonist-sensitive seizures when injected i.c.v. (intracerebroventricular) in mice and produced hippocampal CA1 damage following i.c.v. infusion in rats. In this study, the ability of either 2-ICA, glutamate, proline or NMDA (N-methyl-D-aspartate) injected i.c.v. to produce hippocampal lesions sensitive to glutamate antagonists was compared in mice. Hippocampal CA1 damage was observed 5-days following either a seizure (3.2 mumol) or subseizure (1.0 mumol) dose of 2-ICA. Glutamate (3.2 mumol) or proline (10 mumol) also produced hippocampal damage; glutamate damage was primarily to the CA1 subfield, whereas proline damaged neurons throughout the entire hippocampal formation. NMDA (3.2 nmol) caused seizure activity in all animals with a 50% lethality. No hippocampal damage was observed in surviving mice. Neither MK-801 (dizocilpine maleate) nor CNQX (6-cyano-7-nitroquinoxaline 2,3-dione) pretreatment prevented hippocampal lesions produced by 2-ICA. In contrast, MK-801 significantly reduced the frequency of mice displaying glutamate hippocampal lesions, but failed to block seizures produced by glutamate. MK-801 also protected neurons in the CA2-3 zone and the dentate gyrus, but not in the CA1 region of proline-injected mice. Finally, pretreatment with the mixed metabotropic glutamate receptor (mGluR)1/mGluR2 antagonist-agonist (S)-4-carboxy 3-hydroxyphenylglycine (CHPG) prevented hippocampal damage produced by the mGluR1 agonist (RS)-3,5-dihydroxyphenylglycine (DHPG), but did not protect against 2-ICA hippocampal lesions. These results show that 2-ICA hippocampal CA1 damage is not mediated through ionotropic or metabotropic glutamate receptors. 2-ICA hippocampal damage may represent a neurotoxicity that is distinct from excitotoxic-mediated cell death. PMID- 9216003 TI - Postnatal lead exposure and MK-801 sensitivity. AB - Postweaning Pb exposure has been associated with subsensitivity to the stimulus properties of the non-competitive NMDA receptor complex antagonist MK-801 (Cory Slechta, 1995a). This study sought to determine whether Pb exposures occurring postnatally, i.e., during the primary period of development of many NMDA receptor subunits, would alter the nature of these glutamatergic system changes. Rat pups were exposed to Pb from 0-21 days of age via lactating dams consuming solutions of 0, 100 or 350 ppm Pb acetate. Beginning at 9 mos of age, rats were trained to discriminate 0.05 mg/kg MK-801 from saline using standard operant drug discrimination procedures. Following acquisition of the discrimination, various doses of MK-801, the non-competitive antagonist phencyclidine (PCP), the competitive antagonist 3-(2-carboxypiperazin-4-yl)propyl-1-phosphonate (CPP) and the agonist N-methyl-D-aspartate (NMDA) were substituted for 0.05 mg/kg MK-801 and percent MK-801 lever responding to each determined. Subsequently, a drug washout period was imposed, after which MK-801 dose-effect curves were re established. Increasing doses of MK-801 and PCP produced dose-dependent increases in MK-801 lever responding resulting in full substitution, whereas CPP and NMDA evoked primarily saline-appropriate responding. Pb exposure was associated with enhanced MK-801 sensitivity during the pre-washout phase, but attenuated sensitivity following MK-801 washout. In both cases, however these effects were of relatively modest magnitude. No systematic Pb-related changes in response to PCP, CPP or NMDA were observed. These data raise the possibility, particularly when considered in relation to studies based on other Pb exposure protocols, that NMDA receptor changes may depend upon ongoing or extant Pb exposures, or that postnatal exposure effects on this system may be largely reversible. In addition, the differential nature of the effects seen with postnatal vs postweaning exposure (Cory-Slechta, 1995a) underscores the significance of the developmental period of exposure to Pb effects on the NMDA receptor complex. PMID- 9216004 TI - Effect of long-term lead exposure on hematology, blood biochemistry, and growth curves in monkeys. AB - A total of 90 monkeys (Macaca fascicularis), comprising four study cohorts born over a seven-year period, were hand reared and dosed orally with lead or vehicle according to one of several protocols, in most cases from birth to 9-14 years. Blood lead concentrations of lead-exposed groups ranged from 10 to 90 micrograms/dl depending on dose and age. Routine hematology and blood biochemistry analyses were performed regularly. Comparison of treated groups at various ages to the appropriate control group revealed no strong indication of lead-related effects. In addition, body weight increase was modeled from days 30 3500 of age in subset of this larger group, including 52 monkeys exposed to vehicle or lead during development according to one of four regimens: vehicle, lead from birth onward, lead to 400 days of age, or lead beginning at 300 days of age. No effect of lead on body weight was found. These results suggest that lead exposure beginning early in life and continuing for as long as 14 years resulted in no overt toxicity, as measured by these parameters. PMID- 9216005 TI - Effects of low level lead exposure on cognitive function in children: a review of behavioral, neuropsychological and biological evidence. AB - Low level lead exposure, at levels currently found in significant numbers of children in the U.S., has been associated with decreases in IQ and other cognitive test scores in children, as well as with decreases in developmental test scores in infants. The precise nature of the cognitive deficits is not clear. This paper reviews epidemiological and developmental neurocognitive effects of lead and addresses methodological issues that may have contributed to differences in interpretation of previous research. In an attempt to provide a rationale for the lead-related deficits reported for humans, we have reviewed studies of lead-related behavioral and electrophysiological effects seen in animals as well as findings from studies that have examined the effects of lead exposure on neurochemical subcellular and cellular mechanisms. Based on these data, future strategies are suggested for determining the possible effects of low level lead exposure on neurocognitive functioning in children. PMID- 9216006 TI - Cell culture models of interspecies selectivity to organophosphorous insecticides. AB - In toxicology, the need to reduce uncertainties in human risk assessment is met by understanding why species and individuals within that species respond differently to chemical exposure. This kind of information is needed when extrapolating data from experimental (i.e., whole animal) systems to the human condition in terms of risk assessment. In 1993 the Neurotoxicology Division of the Environmental Protection Agency funded several investigators to examine this phenomenon (i.e., interspecies selectivity) using cell culture models. Organophosphorous (OP) insecticides were examined since they are characterized by an extremely divergent interspecies response. In 1995, a symposium entitled Novel Insights into Chemical Neurotoxicity, sponsored by the Society for In Vitro Biology featured this research. In it, a historical overview of the phenomenon of interspecies selectivity to OP insecticides was given, current explanations for it were discussed and contemporary in vitro models being used to explain it, were described. Data from these studies have helped to redefine the underlying mechanisms that characterize and influence the cross-species response to insecticides. These experiments have refocused the explanation of this phenomenon to include cellular metabolism, target enzyme baseline activities, and receptor mediated electrophysiological and second-messenger events. Several investigators on this panel also reported on the use of subcellular markers (e.g., target esterases, second messengers, ionic fluxes) to differentiate neuropathy-causing OP compounds from acetylcholinesterase inhibitors. After these presentations, technical considerations used in the designed of in vitro neurotoxicity studies were discussed. PMID- 9216007 TI - Dilated, miotic-resistant pupil and laser iridotomy in primary angle-closure glaucoma. AB - We analyzed 22 eyes with primary angle-closure glaucoma that underwent initial laser iridotomy to determine which factors could lead to subsequent trabeculectomy. Twenty-two eyes were divided into two groups: (1) the eyes in which intraocular pressure (IOP) could be controlled by iridotomy and/or topical medication (iridotomy success group, 15 eyes) and (2) the eyes that underwent trabeculectomy to control IOP in spite of a patent opening (iridotomy failure group, 7 eyes). The clinical variables between the two groups were analyzed. Age, sex, visual field defect, presenting IOP and cup/disk ratio were not significantly different between the iridotomy success and failure groups. However, presence of peripheral anterior synechiae (PAS) greater than 50% was found more frequently in the iridotomy failure group as compared with the iridotomy success group (4/7 vs. 1/15, p = 0.02). Dilated, miotic-resistant pupils were observed only in the iridotomy failure group (4/7 vs. 0/15, p = 0.004). PAS greater than 50% and dilated, fixed pupils were observed in these same cases (4 eyes). Our results suggest that laser iridotomy may not be helpful in cases with dilated and miotic-resistant pupils with formation of extensive PAS. PMID- 9216008 TI - Reproducibility of vitreous fluorophotometry in patients with type 1 diabetes mellitus. AB - We assessed the day-to-day reproducible of vitreous fluorophotometry in 7 type 1 diabetic patients and in 1 healthy control subject. The coefficient of variation for duplicate measurements was 33.7, 40.3 and 23.6%, for the right eyes, the left eyes and mean values of both eyes, respectively. Assessment of reproducibility in 1 healthy subject, measured 4 times, resulted in coefficients of variation of about 60%. These values are somewhat worse than those obtained by others in healthy subjects. Since technical problems do not seem to play a significant role in this regard, our reproducibility results reflect the true biological variability of the permeability of the blood-retina barrier assessed by vitreous fluorophotometry in diabetic patients. This variability is substantial. Therefore, we conclude that this method is not precise enough to be used in intervention studies with a limited number of subjects. PMID- 9216009 TI - Contrast sensitivity in patients with silicone intraocular lenses. AB - Contrast sensitivity in 64 patients aged from 50 to 69 years with an intraocular lens (IOL) was studied. There were 23 eyes with a silicone and 31 eyes with a polymethylmethacrylate (PMMA) IOL. Ten patients had a silicone IOL in one eye and a PMMA IOL in the other. Contrast sensitivity was examined with the Vistech far vision VCTS 6500 test. The contrast sensitivity test results in silicone and PMMA IOL eyes were in all cpd (cycles per degree) lines significantly worse than in normal eyes except in the 60-year-old group in the 18-cpd line. The contrast sensitivity test results were in all cpd lines better in the silicone IOL eyes than in the PMMA IOL eyes, but it reached significantly only in the 3-cpd line. In the 10 patients with the silicone lens in one and the PMMA lens in the other eye, the test results were also better in the silicone IOL eye; it also reached significance only in the 3-cpd line. PMID- 9216010 TI - Silicone oil for recurrent vitreous hemorrhage in previously vitrectomized diabetic eyes. AB - AIMS: To investigate the clinical course of vitrectomized patients with recurrent diabetic vitreous hemorrhage who were treated by revitrectomy with silicone oil (SO) as a hemostyptic tamponade. PATIENTS AND METHODS: Fifteen patients with recurrent vitreous hemorrhage due to proliferative diabetic vitreoretinopathy were included in this retrospective study. All eyes had had at least one vitrectomy prior to use of SO and the retina was completely attached at any time before revitrectomy with SO instillation. Thirteen patients had a blind fellow eye. There were 6 males and 9 females (mean age 62.7 years, range 45-76 years). The mean duration of SO tamponade was 25.8 months (range 9-35 months). The average follow-up period was 30.4 months (range 20-48 months). RESULTS: Ten out of 15 eyes (66.6%) improved postoperatively, 9 eyes had a visual acuity of > or = 0.02 at the latest follow-up visit. Secondary glaucoma occurred in 4 eyes, leading to phthisis in 1 eye. All 5 phakic eyes developed a cataract. CONCLUSION: A revitrectomy combined with a long-term hemostyptic SO tamponade offers a chance for restoration of useful visual acuity in diabetic eyes with persistent vitreous hemorrhage that fails to subside after cryocoagulation and vitrectomy without tamponade. Because of possible visual loss from secondary glaucoma related to intraocular SO this treatment should mainly be considered in patients with a blind fellow eye. PMID- 9216011 TI - Role of interleukin 8 in the pathogenesis of proliferative vitreoretinopathy. AB - We investigated the role of interleukin 8 (IL-8) in the pathogenesis of proliferative vitreoretinopathy (PVR). The IL-8 level in the vitreous and the blood of 20 patients with PVR was measured by a specific enzyme-linked immunoassay method. Vitreous from cadaver eyes (n = 20) was used as the control. The IL-8 level in the vitreous was between 31.3 and 65 pg/ml in 40% of eyes with PVR. IL-8 was detected neither in the blood of the patients with PVR nor in the vitreous from cadaver eyes. IL-8 levels in the vitreous did not correlate with either the grade of PVR or the duration of symptoms. These results suggest that IL-8 may play a role in the pathogenesis of PVR. PMID- 9216012 TI - Presence of growth factor in human vitreous. AB - The aqueous humor and the vitreous not only maintain integrity of the eye but also play a vital role in providing nutrients to ocular tissues. While the presence of epidermal growth factor, basic fibroblast growth factor, and transforming growth factor-beta (TGF-beta) have been confirmed in human aqueous humor, only TGF-beta has been found to exist in human vitreous. In this study I obtained eyes for keratoplasty from 20 male and 20 female donors, or a total of 40 subjects; upon removing the corneal button. I extracted 500 microliters of the vitreous humor in order to confirm, using the ELISA method, the presence of transforming growth factor-alpha (TGF-alpha) and epidermal growth factor (EGF) in the vitreous. TGF-alpha ranging from 78 to 250 pg/ml, or an average of 130.65 +/- 40.89 pg/ml, was found in all eyes. On the other hand, EGF was not confirmed in all eyes. This is the first study to confirm the presence of TGF-alpha in the human vitreous. PMID- 9216013 TI - Penetration of oral cefuroxime axetil into the human aqueous humor. AB - The purpose of this study was to investigate the penetration into the aqueous humor of cefuroxime after a single oral dose as cefuroxime axetil. Fourteen patients scheduled for cataract extraction received a single oral dose of cefuroxime axetil corresponding to 500 mg of cefuroxime 2-8 h preoperatively. Aqueous humor samples were obtained at the beginning of the cataract surgery and blood samples were drawn at the time of anesthesia. Cefuroxime levels were determined by high-performance liquid chromatography. The aqueous levels were (mean +/- SEM) 0.48 +/- 0.13 microgram/ml from 3 to 8 h after administration. Serum levels averaged 3.80 +/- 0.58 micrograms/ml. These data indicate that detectable levels of cefuroxime, exceeding the MIC of some bacterial species that frequently cause intraocular infections, may be achieved in uninflamed eyes after a low dose of cefuroxime axetil. PMID- 9216014 TI - Parameter estimation and dosage adjustment in the treatment with vancomycin of methicillin-resistant Staphylococcus aureus ocular infections. AB - The susceptibility of methicillin-resistant Staphylococcus aureus (MRSA) to vancomycin (VC) is excellent, but excessive dosing of VC leads to severe side effects. In this study, we present a way of administration planning for each patient based on personal data. To this end, we employed the Sawchuck-Zaske equation to calculate VC clearance (CL) and volume of distribution (Vd) on the basis of three different plasma concentrations of VC, and we used CL and Vd values as prior information before employing the parameter estimation and dosage adjustment program to treat a patient with postsurgical endophthalmitis caused by MRSA and a patient with acute dacryocystitis also caused by MRSA. They had good remissions without any severe side effects. PMID- 9216015 TI - Cystoid macular edema as an initial symptom of inflammatory orbital pseudotumor. AB - We report a patient with pseudotumor with cystoid macular edema (CME). The initial finding in our case was only CME with a bilateral visual acuity decrease to 20/25. Approximately 3 months later, the visual acuity dropped to light perception in the right eye and 20/200 in the left eye. Computed tomography scan revealed a mass in the right orbital apex and band-shaped enhancement in the cavernous sinus and along the upper margin of the petrous bone. However, no mass was found intraoperatively, and a biopsy specimen of the bulging levator muscle showed polymorphonuclear leukocyte infiltration. The CME resolved postoperatively. The tumor also seemed to resolve; however, after 1 year, the tumor recurred and invaded the brain tissue. A temporal lobectomy revealed widespread inflammatory cell infiltration. To ensure early diagnosis, pseudotumor should be considered in patients with CME of which the cause is uncertain. PMID- 9216016 TI - The impact of early lens opacity progression on visual acuity and refraction. AB - The purpose of the present study was to evaluate the change in visual acuity and refraction taking place in eyes with progressing early lens opacities. Four hundred and ten hypercholesterolemic men in Eastern Finland who participated in the Kuopio Atherosclerosis Prevention Study were followed up for 3 years. Lens opacities were graded using the lens opacity classification system II (LOCS II). The change of visual acuity and refractive error from baseline to a 36-month examination was compared for different types of lens opacities. During the 3-year period, progression in the LOCS II was observed in 9.2% of the eyes for nuclear, in 4.8% for cortical and in 0.5% of the eyes for posterior subcapsular opacities. Increasing nuclear sclerosis reduced visual acuity statistically significantly both with and without correction. Hypermetropization was seen to continue in eyes with no lens opacity progression. Myopization was more common in eyes with lens opacity progression, although this was not statistically significant. PMID- 9216017 TI - Late recurrence of retinal detachment. AB - We reviewed 1,136 cases of retinal detachment in 1,073 patients. Late recurrence was defined as the reappearance of subretinal fluid after at least 6 months of clear, total reattachment. Fifty-one patients (4.75%) had a late recurrence. New or reopened peripheral breaks were either definitely observed or presumed to be present in 40 eyes (78.5%), and none were definite peripheral holes or retinal dialyses. Three macular holes reopened after having initially been managed by vitrectomy. One additional macular hole occurred as a new break. Late severe proliferative vitreoretinopathy occurred in the last 7 cases. Only vitrectomy for initial detachment and lens extraction after reattachment correlated with late recurrence. Vitrectomy was still a significant risk factor when macular holes were excluded. PMID- 9216018 TI - Work-related accidents of ophthalmologic interest in Italy during 1986-1991. AB - The authors analysed accidents of ophthalmological interest obtaining information from the database of the INAIL (National Insurance Institute for Professional Casualties) concerning the period of 1986-1991. The INAIL registers all casualties that cause work disabilities exceeding 3 days. The investigation included all job types grouped into two major categories: agriculture (16% of all insured labour) and industry/craftsmanship (84%). Over 78% of the cases examined were in the industry/craftsmanship category. Approximately 22% of the cases were in the agriculture category. Every year in Italy, about 6% of regularly employed workers suffer casualties. The incidence of casualties of ocular interest has been a stable 0.37% in the years examined; 2.88% of these casualties produce permanent consequences (1/10,000 workers per year). The risk in 3 times higher in agriculture. PMID- 9216019 TI - Etiology of an unusual visual field deficit associated with a craniopharyngioma: case report. AB - We present an unusual case of a craniopharyngioma with a visual field deficit related to optic tract compression by the anterior cerebral artery. The presentation and management of this case are described. Previous cases of visual field deficits associated with craniopharyngiomas are reviewed. PMID- 9216020 TI - Andogsky syndrome and association of other genodermatoses. PMID- 9216021 TI - Psoriatic arthritis or overlap syndrome. AB - In this work we describe a case of papillophlebitis type II according to Hayreh in a young woman affected by psoriatic arthritis. On the basis of an analysis of the patient's HLA system--B7, DR7 (characteristics of psoriatic arthritis) and B51 (present in 81% of patients affected by Behcet's syndrome)--we hypothesize that this may be a case of an 'overlap syndrome'. In rheumatology, this term usually refers to the presence in a single patient of characteristic features of two or more diseases. In fact, papillophlebitis is a complication which has never been described in psoriatic arthritis, while, in Behcet's syndrome, retinal vasculitis is well known. PMID- 9216022 TI - Morphologic changes of the macula in a patient with Purtscher's retinopathy. AB - Purtscher's retinopathy is a rare complication after trauma to the chest or bone fractures. We report about a patient, which we examined 6 months after a serious car accident. Visual acuity was 20/20 in both eyes. Examination with the Amsler grid revealed a paracentral scotoma in the left eye. In the fluorescein angiography of the left eye performed with a scanning laser ophthalmoscope we found capillary dropout, which corresponded well to the scotoma. Measured by digital image analysis, the area of the foveal avascular zone was eccentrically enlarged by a factor of 4. The mean perifoveal intercapillary area was also enlarged in the corresponding quadrant. This reflects that focal capillary dropout may result in scotoma rather than in a decrease in visual acuity as reported in other diseases. PMID- 9216023 TI - Mass blindness has shifted from infection (onchocerciasis, trachoma, corneal ulcers) to cataract. PMID- 9216024 TI - Picture activity schedules and engagement of adults with mental retardation in a group home. PMID- 9216025 TI - Stimulus control and resistance to extinction in attention-maintained SIB. AB - A functional analysis of the self-injurious behavior (SIB) of a young man diagnosed with severe mental retardation demonstrated that SIB was sensitive to social attention as reinforcement. In addition, lower but consistent rates of SIB occurred in sessions where a person was present (Demand and Toy Play), and a gradual decrease in SIB was observed across sessions where a person was not present (Alone). Evaluation of the within-session trends of SIB during the functional analysis demonstrated that SIB maintained throughout each Social Attention session and declined within and across Alone sessions. This pattern of responding suggested that the presence of a person may have differentially affected rates of SIB independent of the programmed consequences for SIB. In a subsequent analysis, SIB was reduced to near-zero levels in the absence of a person, but maintained in the presence of a person even when attention was withheld, suggesting that the response was highly resistant to extinction. The results of these assessments then were used to develop a treatment to reduce the client's SIB. During treatment, a person was present and delivered attention only when the client appropriately communicated. SIB resulted in the removal of the antecedent stimulus that exerted control over the response (i.e., the person left the room). The findings of this investigation are discussed in terms of the differential effects of stimuli on interpretation of functional analysis results and the subsequent development of treatment. PMID- 9216026 TI - Predictors of BMI among adults with Down syndrome: the social context of health promotion. AB - The study explored the relationship of diet, exercise, disability status, and degree of social integration to Body Mass Index, an indicator of excess weight and health status. Subjects were adults with Down syndrome living at home with their families. Variables included a 110-item nutritional analysis and assessments of family demographics, severity of disability, and "lifestyle" variables, such as friendship and affiliation, access to recreation and social activity, and level of physical activity. A factor analysis reduced lifestyle variables into three distinct factors representing friendship, social opportunity, and physical competency. Factor scores were entered into a hierarchical regression model that compared the variance predicted by these factors to the variance accounted for by diet, exercise, and health and physical status variables. Although the overall regression was not statistically significant, the final block of predictors, which represented friendship and social opportunity effects, accounted for a significant increment in BMI variance. Thus, even after the effects of diet, exercise, and physical status variables were partitioned out, the lifestyle variables remained potent predictors of BMI. Study conclusions are described in the context of current paradigms of health in the field of mental retardation and their relationship to inclusion in the community. PMID- 9216027 TI - Analysis and intervention with two topographies of challenging behavior exhibited by a young woman with autism. AB - A functional analysis was conducted with a young woman who engaged in both self injury and aggression. Self-injury functioned to access preferred stimuli while aggression served an escape function. Intervention, a package consisting of gradually increasing the delay to reinforcement (access or escape), mand training, and extinction was effective for decreasing self-injury. However, this intervention was less effective in reducing aggression. A modification of the intervention, in which the gradual delay procedure was eliminated, resulted in reductions in aggression. The findings are discussed in the context of assessment and intervention selection with individuals who engage in multiple topographies of challenging behavior. PMID- 9216028 TI - A comparison of the Diagnostic Assessment for the Severely Handicapped-II (DASH II) and the Aberrant Behavior Checklist (ABC). PMID- 9216029 TI - The strategic approach to contraceptive introduction. AB - The introduction of new contraceptive technologies has great potential for expanding contraceptive choice, but in practice, benefits have not always materialized as new methods have been added to public-sector programs. In response to lessons from the past, the UNDP/UNFPA/WHO/World Bank Special Programme of Research, Development, and Research Training in Human Reproduction (HRP) has taken major steps to develop a new approach and to support governments interested in its implementation. After reviewing previous experience with contraceptive introduction, the article outlines the strategic approach and discusses lessons from eight countries. This new approach shifts attention from promotion of a particular technology to an emphasis on the method mix, the capacity to provide services with quality of care, reproductive choice, and users' perspectives and needs. It also suggests that technology choice should be undertaken through a participatory process that begins with an assessment of the need for contraceptive introduction and is followed by research and policy and program development. Initial results from Bolivia, Brazil, Burkina Faso, Chile, Myanmar, South Africa, Vietnam, and Zambia confirm the value of the new approach. PMID- 9216030 TI - Levels and determinants of gynecological morbidity in a district of south India. AB - This article presents the results of an assessment of gynecological morbidity among 385 women with young children residing in a district of Karnataka State, South India. All three main modes of assessment (clinical examination, laboratory tests, and self-reports) reveal a high burden of reproductive tract infections. The two most common conditions, identified by laboratory tests, were bacterial vaginosis and mucopurulent cervicitis. Approximately one-fourth of the women had clinical evidence of pelvic inflammatory disease, cervical ectopy, and fistula. The contribution of sexually transmitted diseases to overall gynecological morbidity appears to be relatively modest; 10 percent were so diagnosed. Associated conditions of anemia and chronic energy deficiency were common. Severe anemia was found in 17 percent of cases and severe chronic energy deficiency in 12 percent. These results indicate that radical improvements in women's health in India will require far more than the diagnosis and treatment of reproductive tract infections. PMID- 9216031 TI - Estimates and explanations of gender differentials in contraceptive prevalence rates. AB - This article examines gender differentials in the reporting of contraceptive use and offers explanations regarding the sources of these differences. Data from five countries where DHS surveys were conducted recently among men and women are used in exploring these differences. The gap exists in all five countries, with men (or husbands) reporting greater practice of contraception than women (or wives). Results from the bivariate analysis suggest that the gap is attributable to polygyny and to gender differences in how the purpose of contraception is understood, rather than to male extramarital sexual relations. Additionally, gender differences in the definition of certain contraceptive methods and differences in the interpretation of questions about contraception contribute to the observed gap. These findings are also consistent with results of the multivariate analysis. PMID- 9216032 TI - Acceptance, efficacy, and side effects of Norplant implants in four counties in north China. AB - This report attempts to present a comprehensive analysis of the acceptability, side effects, and efficacy of Norplant as used in rural areas, based on a field experiment conducted in four counties in Hebei and Shandong Provinces, China. The initial acceptance of Norplant was relatively high but waned after the first year in three of the four counties. Compared with clinical trials, the current study shows a lower prevalence but similar patterns of side effects. The pregnancy rate during the first two years of use is similar to that found in large-scale clinical trials conducted in China, but discontinuation due to other reasons is lower. A three-level logistic regression analysis shows significant variation in the probability of discontinuation due to side effects across counties. It also indicates an increase in the conditional probability of discontinuation with the duration of use. Whereas introducing Norplant and achieving a very low failure rate and high continuation rate in rural areas is feasible under diverse socioeconomic conditions, the results vary significantly across different areas. Particular attention should be paid to the local factors that may affect results. PMID- 9216033 TI - Accuracy of indirect estimates of maternal mortality: a simulation model. AB - A simulation model was developed to test the accuracy of indirect estimates of maternal mortality (the sisterhood method). The model generated a first generation of grandmothers, a second generation of mothers (with brothers and sisters), and a third generation of children (births). In the second generation, maternal mortality was introduced. Empirical values for the parameters of fertility and mortality were taken from a prospective survey in Senegal (Niakhar). Results based on 100 simulations of the same situation revealed several limitations of the sisterhood method: The indirect estimates could fall as far as 33 percent from the true values on individual cases; the indirect estimates tended to be systematically higher than the direct estimates; their range was wider, as were their confidence intervals; and biases were particularly strong for the younger age groups of respondents. Reasons for these biases are explored. PMID- 9216035 TI - Guatemala 1995: results from the Demographic and Health Survey. PMID- 9216034 TI - Assessing family planning service-delivery skills in Kenya. AB - This report demonstrates the use of Lot Quality Assurance Sampling (LQAS) to evaluate the technical competence of two cohorts of family planning service providers in Kenya trained with a new curriculum. One cohort had just finished training within two months of the study. The other cohort was the first group trained with the new curriculum about one year before the study. LQAS was adapted from industrial and other public health applications to assess both the individual competence of 30 service providers and the competence of each cohort. Results show that Cohorts One and Two did not differ markedly in the number of tasks needing improvement. However, both cohorts exhibited more tasks needing improvement in counseling skills as compared with physical examination skills or with all other skills. Care-givers who were not currently providing services accounted for most service-delivery problems. This result suggests that providers' use of their skills explains their ability to retain service-delivery skills learned in training to a greater degree than does the amount of time elapsed since they were trained. LQAS proved to be a rapid, easy-to-use empirical method for management decisionmaking for improvement of a family planning training curriculum and services. PMID- 9216036 TI - Uganda 1995: results from the Demographic and Health Survey. PMID- 9216037 TI - On evaluating mass media's impact. PMID- 9216038 TI - Midkine and secondary neurulation. AB - Midkine (MK) is a growth factor with documented neurotrophic activity for central nervous system neurons. It has also been shown to induce conversion of pluripotent mesenchymal P19 embryonic carcinoma cells into neuroepithelial derivatives. During the development of the chick embryonic neuraxis, two separate events occur. The anterior, greater portion of the neural tube, the primary neural tube, develops first from the neuroectoderm. The posterior section of the neural tube or the secondary neural tube forms next, from the cells of the tail bud, as tail bud cells undergo mesenchymal-neuroepithelial conversion. Disruptions in tail bud differentiation lead to defects of the secondary neural tube. In this study, antisense oligodeoxynucleotides (ODNs) were used to inhibit MK expression in order to determine if MK has a role in the mesenchymal neuroepithelial conversion process. Chick embryos at the tail bud anlagen stage were treated with antisense ODNs to MK mRNA by sub-blastodermal injection and reincubated for 24 or 48 hr. The antisense ODN treatment resulted in a significant increase in the incidence of secondary neural tube defects, compared to control treatments with the saline vehicle only, sense ODNs, scrambled antisense ODNs, or non-sense ODNs. The loss of MK mRNA in the antisense ODN treated embryos was confirmed by in situ hybridization. The results therefore suggest a role for MK in the mesenchymal-neuroepithelial conversion of tail bud mesenchyme into the secondary neural tube. PMID- 9216039 TI - Heterogeneity of spina bifida. AB - Splitting birth defects into dysmorphologically homogeneous groups might improve the ability to detect a genetic risk factor or teratogenic exposure. With regard to spina bifida, recent studies suggest that etiologic heterogeneity exists within the group of spina bifida, although exogenous risk factors have been sparsely evaluated for subgroups. In the present study, 210 spina bifida patients were classified into relatively homogeneous groups, based on retrospective information on appearance and functional aspects of the lesion abstracted from medical records of the patients. We compared high with low spina bifida, and open with closed spina bifida, and investigated whether risk factors for spina bifida such as maternal age, antiepileptic drug use, parental occupation, and genetic factors were specifically associated with these homogeneous subclasses. For these comparisons, a referent group of 671 children was used. Although classifying spina bifida into homogeneous subclasses presented some difficulties and numbers were small, this study provides some evidence for different risk profiles for subclasses of spina bifida. The sex ratio, the proportion of miscarriages of siblings, and maternal age did not differ among the different subclasses of spina bifida. However, children with a positive family history of neural tube defects (NTDs) had a higher risk of high spina bifida [odds ratio (OR) = 6.3, 95% confidence interval (CI): 1.7-19.2] than of low spina bifida (OR = 2.1, 95% CI: 1.0-4.2). Siblings with NTDs were more common in cases with high spina bifida and cases with open spina bifida. A strongly increased risk of high spina bifida was found for male welders (OR = 12.1, 95% CI: 1.5-64.2), whereas the risk of low spina bifida was much lower (OR = 1.6, 95% CI: 0.2-7.9). For mothers with agricultural occupations, a strongly increased risk was observed for open spina bifida (OR = 14.3, 95% CI: 2.9-77.7), whereas none of 107 cases with closed spina bifida had a mother with an occupation in agriculture. Due to small numbers, the results must be interpreted with caution. PMID- 9216041 TI - Vertebral column and spinal cord malformation in children with exstrophy of the cloaca, with emphasis on their functional correlates. AB - Exstrophy of the cloaca is a dramatic malformation whose embryology is poorly understood. While the management of this disorder has received significant attention in the urology and general surgery literature, the neurologic status of these children has been poorly addressed. In order to better characterize the spinal cord and vertebral column malformations found in children with exstrophy of the cloaca, we undertook a clinical review of 26 consecutive children with exstrophy of the cloaca who had been seen at a single institution over 28 years. The prevalence of vertebral malformations in the 25 children who could be evaluated was 25/25 (100%). Twenty (80%) of the children had at least one vertebral fusion, most frequently at T-7. Twenty-two (88%) of the children had at least one vertebra with deficient posterior elements, and the spinal levels most frequently involved were S-2, S-3, S-4 and S-5. Nine (36%) of the children had at least one vertebra with a narrowed interpedicular distance, most frequently at T 7. Nine (36%) of the children had at least one vertebra with atrophic facet anatomy, most frequently at L-3. The prevalence of myelodysplasia in the 19 children for whom spinal magnetic resonance imaging or intraoperative findings were available was 100%. Of these 19 children, 15 (79%) had myelocystocele, 2 (11%) had a lipomeningocele, 2 (11%) had a meningocele, 2 (11%) had hydromyelia, and 4 (21%) had a tethered cord. These data suggest that spinal cord and vertebral column malformations are very common in children with exstrophy of the cloaca. PMID- 9216040 TI - Forebrain overgrowth (fog): a new mutation in the mouse affecting neural tube development. AB - Forebrain overgrowth, fog, is a spontaneous autosomal recessive mutation in the mouse producing forebrain, lumbo-sacral, and facial defects. The defects appear to result from excessive growth or cellular proliferation leading to abnormalities in neural tube closure. Three unique features of the mutant are: (1) the growth of telencephalon cells into the surrounding mesenchyme, (2) presence of an encephalocele through the midline cleft in some mutants, and (3) dissociation of the tail defect from the caudal neural tube defect. We used an intersubspecific intercross between mice carrying the fog mutation and mice from an inbred Mus musculus castaneus strain (CAST/Ei) to map the fog mutation to mouse Chromosome 10 near D10Mit262 and D10Mit230 in a region with several potential candidate genes. PMID- 9216042 TI - Terminology of developmental abnormalities in common laboratory mammals (version 1). AB - This paper presents the first version of an internationally-developed glossary of terms for structural developmental abnormalities in common laboratory animals. The glossary is put forward by the International Federation of Teratology Societies (IFTS) Committee on International Harmonization of Nomenclature in Developmental Toxicology, and represents considerable progress toward harmonization of terminology in this area. The purpose of this effort is to provide a common vocabulary that will reduce confusion and ambiguity in the description of developmental effects, particularly in submissions to regulatory agencies worldwide. The glossary contains a primary term or phrase, a definition of the abnormality, and notes, where appropriate. Selected synonyms or related terms, which reflect a similar or closely related concept, are noted. Nonpreferred terms are indicated where their usage may be incorrect. Modifying terms used repeatedly in the glossary (e.g., absent, branched) are listed and defined separately, instead of repeating their definitions for each observation. Syndrome names are generally excluded from the glossary, but are listed separately in an appendix. The glossary is organized into broad sections for external, visceral, and skeletal observations, then subdivided into regions, structures, or organs in a general overall head to tail sequence. Numbering is sequential, and not in any regional or hierarchical order. Uses and misuses of the glossary are discussed. Comments, questions, suggestions, and additions from practitioners in the field of developmental toxicology are welcomed on the organization of the glossary as well as on the specific terms and definitions. Updates of the glossary are planned based on the comments received. PMID- 9216043 TI - Anatomy and physiology of the bull's reproductive system. AB - The control of bovine male reproduction is a finely orchestrated system. However, the mechanism has already been predetermined by the bull's genetic makeup and, to a lesser extent, the environment. Although the endocrine system appears robust, excessive pressures can inhibit or arrest the process. Similarly, structural limits to the bull's anatomy exist, and exceeding them can result in permanent damage. Hence, a thorough understanding of the anatomy and physiology allows successful management of the breeding bull. PMID- 9216044 TI - Common causes of infertility in the bull. AB - Fertility is a fragile parameter that may vary temporarily or be permanently depressed. The bull's ability to maintain body condition and the existence of conformational abnormalities may alter fertility. Abnormalities of the external genitalia may directly affect the bull's ability to copulate or may alter the quality of his semen. Testicular disease alters semen quality, sometimes transiently and other times permanently. In many cases, the fertility prognosis for testicular problems cannot be judged on the basis of one examination. Epididymitis and seminal vesiculitis are the two most common diseases of the secondary sex organs of the bull. They are often associated with each other or orchitis and are difficult to treat. PMID- 9216045 TI - Bovine seminal vesiculitis. A review and update. AB - Seminal vesiculitis is the most common inflammatory condition affecting the reproductive tract of the bull. It represents a serious source of economic loss. This ailment is frequently seen in young peripubertal bulls and occasionally in older bulls, and it can negatively affect semen quality. Multiple etiologic agents have been cultured from seminal vesiculitis cases. Medical therapy is often unsuccessful; however, surgery offers some hope for selected young bulls. PMID- 9216046 TI - Genetic causes of bull infertility. AB - Infertility and anatomic defects unique to breeding bulls can be influenced by genetics. Veterinarians and animal breeders need to report disorders that may be inherited to a central recording agent, usually the breed association or a veterinary specialist interested in characterizing the disease. Occurrence of these defects is rare, and a large population of animals should be studied to recognize patterns of inheritance early. Numbers of affected animals in a given practice area are often limited, leading to an underestimation of the condition's importance. Genetic defects occur regularly at low frequencies. New syndromes continually arise and most are silently eliminated. Cooperation between cattle breeders, veterinarians, breed associations, and the scientific community is essential in controlling these diseases. PMID- 9216047 TI - The significance to bull fertility of morphologically abnormal sperm. AB - Bull fertility depends partially on the quality of the spermatozoa (i.e., different spermatozoa defects) produced in the ejaculate, emphasizing the importance of the interpretation of the spermeogram results. This article discusses the production of normal and abnormal spermatozoa and the influence that spermatozoa defects may have on bull fertility. PMID- 9216048 TI - Scrotal/testicular thermoregulation and the effects of increased testicular temperature in the bull. AB - Scrotal/testicular thermoregulation is a complex process controlled by numerous local mechanisms that attempt to maintain the testes at conditions ideal for spermatogenesis. This article provides a background of the anatomy and physiology of the bovine scrotum and its contents with emphasis on thermoregulation. Experiments are cited that demonstrate scrotal/testicular thermoregulation mechanisms and the effect that changes in ambient temperature have on internal testicular temperature and subsequent seminal quality. PMID- 9216049 TI - The new Society for Theriogenology breeding soundness evaluation system. AB - In 1993, The Society for Theriogenology published revised standards for breeding soundness evaluation of bulls. These revised standards replaced the old semen score format with minimum acceptable scores for scrotal circumference (varies with age), motility (30% or fair), and morphology (70%). PMID- 9216050 TI - Breeding soundness evaluation of yearling bulls. AB - Breeding Soundness Evaluations (BSEs) are a useful tool to improve reproductive performance and profitability in beef herds. Veterinarians providing this service must be thorough in their evaluation, especially in yearling bulls. The 1993 Society for Theriogenology guidelines for BSE provide appropriate standards for classifying yearling bulls; veterinarians are advised to adhere to these standards as minimums for classification of the satisfactory potential breeder. PMID- 9216051 TI - Observations using the new bull-breeding soundness evaluation forms in adult and young bulls. AB - The new bull-breeding soundness forms were used to evaluate 1276 bulls; 62.85% of these bulls were deemed satisfactory. Unsatisfactory and deferred percentages were 28.92% and 8.23% respectively. Reasons for the various classifications were evaluated, and age and breed comparisons were made. PMID- 9216052 TI - Ancillary tests of bull semen quality. AB - Seminal quality examinations readily identify animals with low fertility, but rarely can these tests discriminate among males with moderate to high fertility. The new, automated semen analysis systems tend to be helpful in providing more reliable information. Such systems, especially the CASA systems that assess sperm morphology and motility, are innovative and certainly attractive. They may not, however, be cost effective. Similarly, although the combination of fluorescent staining and flow cytometry offers a very rapid and precise means of assessing the functional status of sperm organelles, it tends to be relatively expensive as a clinical tool. Such automated sperm quality assessments will likely become routine analyses at the larger semen-processing organizations. However, the true success of a breeding program can only be assessed by the number of live offspring. Thus, determination of true fertility, because of the complexity of the processes that unite the gametes, will continue to require more than a detailed examination of seminal cytology, no matter how sophisticated our methods become. PMID- 9216053 TI - Bull libido/serving capacity. AB - Bull libido, or sex drive, is a measurable trait with a large genetic component. It also represents an important aspect of bull reproductive performance, with positive effects on herd pregnancy rates and their patterns. Comparative, quantitative assessment of this trait requires a formalized testing procedure such as a test for libido or serving capacity score. Such tests are useful not only for obtaining quantitative information, but also for the detection of physical and pathologic problems that may interfere with normal bull mating ability. These tests should be conducted in such a manner that animal welfare is not unnecessarily compromised. The quantitative results should be interpreted with caution, especially when young, inexperienced bulls or those of Bos indicus breeds are being assessed. Libido does not necessarily work in concert with other traits known to separately influence bull fertility (e.g., BSE traits and social dominance). Until a single procedure is found that can adequately assess all of these factors, optimal bull appraisal requires separate evaluation of each of these factors. PMID- 9216054 TI - Pathogenesis, diagnosis, and management of trichomoniasis in cattle. AB - Trichomoniasis is a disease of the pregnancy, but apparently not of either the cow or the bull, except in the case of postcoital pyometra. Its self-limiting nature in the cow and chronic nature in the bull mean that a positive diagnosis for the herd can more easily be obtained from bulls than from cows. Incubation of preputial scrapings or washings (or pyometritic fluid, if available) in a selective growth medium such as the InPouch system is the diagnostic method of choice. The diagnosis is based on identification of the morphology and characteristic rolling motility of the trichomonad. "High tech" molecular approaches may eventually offer greater diagnostic sensitivity than can culture methods, but currently they are no more accurate. In addition, serologic screening of the female herd (but interestingly, not the bulls) may become possible and may allow the practitioner to at least determine whether exposure has occurred in an unvaccinated herd. Control in an infected herd involves no pharmacologic treatment but rather culling of infected bulls, retention of younger, culture-negative bulls, and segregation of the female herd by reproductive status. PMID- 9216055 TI - Group, assimilation, and increase in visibility association without a difference in features. AB - There is evidence that one group is associated with assimilation among its parts and an increase in visibility (IV) of at least one of its parts: the 1 group assimilation-IV position. The present research supports this position using physically identical stimuli, hence eliminating differences in features. This was accomplished by comparing the effects of large and small backgrounds on responding to physically identical stimuli that appeared on these backgrounds. Compared to the small background, the large background produced a stronger two line group according to a closure measure of grouping, more assimilation between two lines according to a same-different measure of perceived similarity, and a greater IV of one of two lines according to context+target versus context relative to target versus background discriminations. The large background was much larger than the two small lines, suggesting that it functioned as an anchor. PMID- 9216056 TI - Effects of cognitive ability and affect on school mathematics performance and feelings of difficulty. AB - Research on cognitive ability and affect has indicated that both of them may function at various levels of generality (LG). This study aimed to investigate the possible effects of LG on school mathematics performance and related feelings of difficulty. Two hundred forty-three students of seventh, eighth, and ninth grades of both genders participated in the study. They were tested with cognitive ability tasks, affective questionnaires, and two school mathematics batteries. The difficulty of each of the school mathematics task was also rated on a 4-point scale. The two school mathematics batteries and affective questionnaires were re administered 1 year after the first testing. Rates of difficulty were also taken. Path analysis showed that performance was mainly influenced by cognitive ability factors, whereas feelings of difficulty were influenced by performance, cognitive ability, and affective factors. The long-term relations between cognitive and affective factors are discussed. PMID- 9216057 TI - Scoring procedure, performance factors, and magnitude of sex differences in spatial performance. AB - The purpose of the present study was to demonstrate the influence of procedural factors on the magnitude of sex differences in a test of spatial ability. Two hundred and seven females and 155 males were administered the Mental Rotations Test (MRT) under timed and untimed conditions. Four different scoring procedures were used: total score, ratio of correct responses to the number of items attempted, score out of 24, and score out of 48. Significant sex differences were obtained in the timed condition but not in the untimed condition. Results also revealed that the magnitude of sex differences was reduced when a ratio score was used. Analysis of the pattern of responses provided insights into the causes of sex differences on the MRT. Results are interpreted in terms of their implications for research on sex differences in spatial ability. PMID- 9216058 TI - Perspectives on eye development. AB - The lens of the vertebrate eye was the classic model used to demonstrate the concepts of inductive interactions controlling development. However, it is in the Drosophila model that the greatest progress in understanding molecular mechanisms of eye development have most recently been mode. This progress can be attributed to the power of molecular genetics, an approach that was once confined to simpler systems like worms and flies, but is now becoming possible in vertebrates. Thus, the use of transgenic and knock-out gene technology, coupled with the availability of new positional cloning methods, has recently initiated a surge of progress in the mouse genetic model and has also led to the identification of genes involved in human inherited disorders. In addition, gene transfer techniques have opened up opportunities for progress using chick, Xenopus, and other classic developmental systems. Finally, a new vertebrate genetic model, zebrafish, appears very promising for molecular studies. As a result of the opportunities presented by these new approaches, eye development has come into the limelight, hence the timeliness of this focus issue of Developmental Genetics. In this introductory review, we discuss three areas of current work arising through the use of these newer genetic approaches, and pertinent to research articles presented herein. We also touch on related studies reported at the first Keystone Meeting on Ocular Cell and Molecular Biology, recently held in Tamarron Springs, Colorado, January 7-12, 1997. PMID- 9216059 TI - Late proliferation of retinal Muller cell progenitors facilitates preferential targeting with retroviral vectors in vitro. AB - During vertebrate neural retina development, the relationship between mitotic activity in progenitor cells and the acquisition of a mature cell phenotype remains an area of controversy. The Muller glial cell has long been recognized as one of the last cell types of the retina to mature, which occurs under the influence of cell-cell interactions. In this report we examine the acquisition of the Muller cell phenotype in relation to mitotic activity. Using immunohistochemical markers, we demonstrate that a gene product characteristic of mature Muller cells, the 2M6 antigen, is expressed in mitotically active cells, even after all the major retina architectural features have been laid down. Furthermore, we show that retroviral infection, a process that requires mitotically active cells, preferentially targets Muller cell progenitors when late embryonic retina is infected in vitro. The two lines of evidence are consistent with a model for Muller cell differentiation that includes a mitotically active progenitor that has already begun to express specific differentiation gene products. PMID- 9216060 TI - Retinal morphogenesis in Drosophila: hints from an eye-specific decapentaplegic allele. AB - Decapentaplegic (dpp) regulates many aspects of imaginal disc growth and patterning in Drosophila. We have analyzed the phenotype of an eye-specific dpp allele, dppblk, which causes a reduction in the size of the retina due to a loss of ventral ommatidia. Prior to the onset of differentiation, dppblk eye discs are normal regarding size, shape, and ability to express dorsal and ventral markers. However, expression of a dpp-lacZ reporter is reduced at the ventral margin. Additional dorsoventral asymmetry appears during retinal differentiation: the morphogenetic furrow (MF) initiates normally at the posterior tip of the disc, but fails to propagate into the ventral epithelium. This defect can be rescued by increasing dpp expression along the ventral margin by local removal of patched function. We propose that the primary defect in dppblk is an inability to activate dpp expression properly at the ventral margin. This has two consequences: it prevents initiation from the ventral margin, and it renders the ventral epithelium unresponsive to differentiation signals emanating from the MF. PMID- 9216062 TI - Evidence that tyrosine phosphorylation regulates N-cadherin turnover during retinal development. AB - N-cadherin, a member of the cadherin family of calcium-dependent cell adhesion molecules, mediates adhesive and signaling interactions between cells during development. N-Cadherin undergoes dynamic spatiotemporal changes in expression which correlate with morphogenetic movements of cells during organogenesis and histogenesis. We have previously shown that N-cadherin expression during development is regulated by several mechanisms, including mRNA expression, cytokine modulation, and proteolytically mediated turnover, yielding the NCAD90 protein. The present study was directed at determining the extent to which N cadherin in primary embryonic cells is the target of endogenous kinases and phosphatases, as well as the effects of modulation of these enzymes on NCAD90 expression. The results of phosphoamino acid analyses, peptide mapping, and measurements of N-cadherin and NCAD90 expression in embryonic tissues indicate that N-cadherin is indeed the target of endogenous kinase and phosphatase action, and that modulation of different classes of these enzymes can result in either stimulation or inhibition of NCAD90 production. These results provide a mechanistic explanation for observations that cadherin function is downregulated following expression of exogenously introduced viral tyrosine kinases and provide a function for the tyrosine phosphatases recently found in association with cadherins. The results indicate that N-cadherin expression during retinal development is possibly regulated in part by modulation of its phosphorylation state, the balance of which may determine whether N-cadherin remains stably expressed or is targeted for proteolytically mediated turnover to produce NCAD90. PMID- 9216061 TI - A goldfish Notch-3 homologue is expressed in neurogenic regions of embryonic, adult, and regenerating brain and retina. AB - Members of the Notch gene family are thought to be involved in the regulation of cell fate decisions in a variety of embryonic tissues, particularly in the developing central nervous system (CNS) in Drosophila and vertebrates. In goldfish the CNS continues to develop and add neurons well into adulthood and has the capacity to regenerate new neurons. Using probes derived from Xenopus Notch to screen an adult goldfish retinal cDNA library, followed by 5' RACE, we isolated a partial cDNA for a goldfish Notch homologue, G-Notch. Sequence alignment supported assignment of G-Notch to the Notch-3 class. Northern blot analysis revealed a single transcript of > 8 kb, and RNase protection assays indicated that G-Notch is expressed in eye and brain but not muscle of adult goldfish. The spatiotemporal pattern of expression of G-Notch was defined from early embryonic stages to adulthood by in situ hybridization. Expression in the embryonic CNS was localized to neurogenic regions and was downregulated in differentiated cell populations. In adult goldfish, expression persisted in and adjacent to the germinal zones in the retina and the brain. Weak expression was seen in scattered cells in the inner nuclear layer of the retina, which might include neurogenic stem cells. Following retinal lesions (puncture wounds or laser lesions restricted to photoreceptors in the outer nuclear layer), G-Notch was upregulated in proliferating cell populations throughout the retina, in association with a generalized mitogenic response. In the region of the laser lesion, where earlier studies have demonstrated that photoreceptors are regenerating at 1-3 weeks following the lesion, G-Notch expressing cells were abundant in the outer nuclear layer. These observations suggest that retinal regeneration involves the re-expression of an important developmental signaling molecule in neuroepithelial cells resident in the differentiated retina. PMID- 9216063 TI - Cloning and characterization of human homologue of Drosophila retinal degeneration B: a candidate gene for degenerative retinal diseases. AB - Mutations in the Drosophila retinal degeneration B (D-rdgB) gene cause light enhanced retinal degeneration. Here, we report the isolation of the cDNA encoding human homologue of the D-rdgB and initial characterization of the gene products. Like D-rdgB, the human rdgB homologue (H-rdgB) is a transmembrane protein with the N-terminus sharing high homology to two closely related cytosolic proteins, phosphatidylinositol transfer protein (PITP) alpha and beta, indicating that rdgB like proteins belong to the family of PITP proteins. Using Northern and Western blotting, we demonstrated that the rdgB homologue is expressed in rat retina, olfactory bulb, and brain, but not in nonneuronal tissues. In the rat retina, immunoreactivity of the rdgB homologue was observed in photoreceptors and throughout the inner nuclear and plexiform layers; the strongest staining was in the inner plexiform layer. In the photoreceptor cells, the rdgB homologue was located primarily in the inner segment where sorting and traffic of membranes required for outer segment assembly take place. These data, together with recent findings showing PITPs as on important component of intracellular membrane traffic apparatus in mammalian cells, suggest that rdgB homologue may play a role in photoreceptor membrane renewal and in neurotransmitter release. Furthermore, using somatic hybrid cell hybridization and fluorescence in situ hybridization H rdgB gene was mapped to human chromosome 11q13, a region known to contain several retinopathy loci, including Best disease and Bardet-Biedl syndrome I. Therefore, H-rdgB gene is an attractive candidate for several inherited retinal degenerative diseases. PMID- 9216064 TI - Defining intermediate stages in cell determination: acquisition of a lens-forming bias in head ectoderm during lens determination. AB - Cell determination in vertebrates requires integration of many events, although the mechanisms controlling the different stages in this process are poorly understood. While studies of lens determination have helped define some of these stages, we know very little about the intermediate steps involved in the commitment of ectoderm to lens formation. Lens determination begins during gastrulation when ectoderm is briefly competent to respond to lens-inducing signals and progresses to a point, at the neural tube stage, when the presumptive lens ectoderm is specified. Between these two stages important regulatory genes are activated in the presumptive lens ectoderm, including the transcription factor Pax-6, and transplantation experiments presented here suggest that the presumptive lens ectoderm is becoming "biased" toward lens formation. We show that competent ectoderm from Xenopus laevis embryos forms well-differentiated lenses in most cases when transplanted to the presumptive lens area of neural plate stage hosts, where the lens-inductive environment is strong. When placed into later, neural tube stage hosts, optimally competent ectoderm does form small lenses in about half the cases, but the overall response is much weaker. Even in this weakly inducing environment, however, lens ectoderm that is part way through the inductive process (at the neural plate stage) is shown to have a lens-forming bias, since it forms well differentiated lenses in nearly all cases. While we find that ectoderm surrounding the lens-forming area at neural plate stages does not have a lens-forming bias, non-lens head ectoderm at the neural tube stage does, suggesting that a large region of head ectoderm is biased during neurulation. Using Rana palustris embryos, a species used in the earliest lens induction studies, we were also able to show that the optic vesicle can induce lenses in non-lens head ectoderm at neural tube stages. These results lead us to refine the definition of lens cell determination and provide a context that should allow clarification of determination mechanisms. PMID- 9216065 TI - Lens-preferred activity of chicken delta 1- and delta 2-crystallin enhancers in transgenic mice and evidence for retinoic acid-responsive regulation of the delta 1-crystallin gene. AB - There are two tandemly linked delta-crystallin genes [5' delta 1 -delta 2 3'] in the chicken, with the delta 1-crystallin gene being expressed much more highly (50-100-fold) in the embryonic lens than the delta 2-crystallin gene. Previous transfection experiments have shown that a lens-preferred enhancer exists in the third intron of each chicken delta-crystallin gene. In the present investigation we have used transgenic mice to establish that both the chicken delta 1- and delta 2-crystallin enhancers are preferentially active in the mouse lens in combination with their homologous promoter and the chloramphenicol acetyltransferase (CAT) reporter gene. The promoter/ CAT constructs lacking the enhancers were inactive in the transgenic mice. In one case, a truncated delta 2 crystallin promoter (-308/+24) in combination with the enhancer was also active in the Purkinje cells of the cerebellum of the transgenic mice, which could prove useful in future experiments. Finally, retinoic acid receptors (RAR beta) activated the delta 1-crystallin, but not the delta 2-crystallin enhancer in teh recombinant plasmids in cotransfected embryonic chicken lens epithelial cells treated with retinoic acid. This activation did not occur when using the care enhancer (fragment B4) lacking surrounding flanking sequences (fragment B3 and B5) of the enhancer. Together these experiments show that the chicken delta crystallin enhancers show lens-preference in transgenic mice despite the absence of delta-crystallin in this species and add retinoic acid nuclear receptors to the growing list of transcription factors (including Pax-6, Sox-2, and delta EF3) that directly or indirectly contribute to the high expression of the delta 1 crystallin gene in the lens. PMID- 9216066 TI - Expression of Cdk5, p35, and Cdk5-associated kinase activity in the developing rat lens. AB - We have investigated the expression of Cdk5 and its regulatory subunit, p35, in the developing rat lens from embryonic day 16 (E16) to postnatal day 8 (P8). Reverse transcription and polymerase chain reaction (RT/PCR) detected Cdk5 and p35 mRNA expression in lens epithelial cells and in differentiating lens fibers throughout this developmental period. Subsequent sequencing of the RT/PCR products confirmed their identifies. In sity hybridization with Cdk5 and p35 riboprobes showed especially high expression of both mRNAs in the newly formed lens fiber cells in the bow region of the lens. Immunocytochemistry at E18 showed that Cdk5 was present in the cytoplasm of lens epithelial cells and fiber cells, with especially strong immunostaining at the anterior ends of the fibers. Fiber cells in the final stages of maturation, immediately prior to nuclear degeneration, showed positive staining for Cdk5 in the nucleus. Immunoprecipitation of proteins with Cdk5 antibody followed by immunoblotting with either N-terminal specific or C-terminal specific p35 antibodies demonstrated that p35 is complexed with Cdk5 in lens epithelial cells and lens fibers. Immunoprecipitates of Cdk5 from epithelia and fibers showed kinase activity in vitro using histone H1 as a substrate. These findings demonstrate that p35/Cdk5 activity is not restricted to neurons and raise the possibility that this kinase may play a role in lens fiber cell differentiation. PMID- 9216067 TI - Inhibition of cell death by lens-specific overexpression of bcl-2 in transgenic mice. AB - Previous studies on cell cycle regulation in the ocular lens using transgenic mice have shown that inactivation of the retinoblastoma tumor suppressor protein (pRb) can cause postmitotic lens fiber cells to enter the cell cycle. However, when the p53 gene and protein are intact, inactivation of pRb in this terminally differentiated cell type results in cell death, rather than continued proliferation. Since bcl-2 has been shown to act as a cell death repressor, the ability of this gene to block p53-dependent apoptosis in lenses was examined. Transgenic mice were generated that overexpress bcl-2 in a lens-specific fashion. Surprisingly, overexpression of bcl-2 was sufficient to interfere with normal fiber cell differentiation, inducing cataracts, microphakia, vacuolization, fiber cell disorganization, and inhibition of fiber cell denucleation. The bcl-2 mice were mated to mice exhibiting lens-specific expression of the N-terminal region of simian virus 40 large T antigen (termed truncT). The resulting double transgenic mice showed a marked reduction in the truncT-induced fiber cell death. Apoptosis in the truncT mice could also be suppressed by crossing these mice into a p53-deficient background. Either overexpression of bcl-2 or loss of p53 in truncT mice resulted in proliferation of fiber cells around the cortex of the lens. These proliferating fiber cells continue to express beta- and gamma crystallin proteins, which are normally only expressed following withdrawal from the cell cycle. The p53 protein is known to upregulate expression of certain target genes, including p21, a protein that can block cell cycle progression by inhibition of cyclin-dependent kinases. In order to assess whether bcl-2 interferes with the transcriptional activation activity of p53, transgenic lenses were assayed by in situ hybridization for levels of p21 expression. Lenses that expressed both truncT and bcl-2 showed elevated p21, implying that bcl-2 does not inhibit apoptosis by directly inhibiting p53, but instead may block a later step in the apoptosis pathway. In addition, overexpression of p21 is not sufficient to cause apoptosis. These experiments show that the lenses of transgenic mice represent a valuable in vivo setting for studies of both induction and inhibition of programmed cell death. PMID- 9216068 TI - Mutations affecting eye morphology in the developing zebrafish (Danio rerio). AB - The zebrafish (Danio rerio) has received considerable attention as a mainstream model for the molecular and genetic study of vertebrate development. In our laboratory, we have conducted a third-generation screen of chemically mutagenized zebrafish for recessive mutations affecting the visual system. This report describes the visible phenotypes and number of morphological mutants so far observed and presents a more detailed histological analysis of six of these mutations. Through analysis of mutant larvae, it was determined that several of the subtle morphological mutations resulted in degeneration of specific cellular layers of the retina. Other mutations resulted in some degeneration distributed diffusely across the entire retina or concentrated at the retinal margin. A single mutation affecting invagination of the optic cup and lens vesicle formation resulted in a failure to develop an anterior chamber. These results demonstrate the utility of a small-scale, highly focused screen for uncovering novel loci involved in retinal and eye development. PMID- 9216069 TI - The effect of superinfection on the distribution of the infectious period--a continued fraction approximation. AB - It has been shown that the density function of the duration of infection in a superinfection malaria model can be approximated by a mixture of exponential density functions. This is achieved by an application of continued fractions. Numerical calculations and graphs illustrate that this approach is effective. PMID- 9216071 TI - The King Solomon Lectures in Neuroethology. Deciding about motion: linking perception to action. PMID- 9216070 TI - Estimation of glomerular filtration rate in type II (non-insulin dependent) diabetes mellitus patients. AB - The aim of this research was to develop an estimation of glomerular filtration rates (GFRs) from a combination of simple parameters in a large group of type II diabetic patients. We selected 122 newly presenting, previously untreated, type II patients whose GFR was determined from the plasma clearance of 51Cr ethylenediamine tetraacetic acid (51Cr-EDTA) and simultaneous measurements of demographic variables, including fasting plasma glucose concentration, HbA1c, blood pressure, lipids, age, weight, body-mass index, body surface area, urea, and plasma creatinine concentration. The actual GFR values were compared with estimated values obtained from multiple regression and the Cockroft-Gault equations. Out of all the demographic variables, only plasma creatinine concentration (r = -0.56, p < 0.001), age (r = -0.50, p < 0.001), urea (r = 0.28, p < 0.01), and systolic blood pressure (r = -0.21, p < 0.05) showed significant correlations with the actual GFR values, for which the mean and standard deviation were 117.5 +/- 22.0 ml min-1 x 1.73 m-2. The estimated values are highly correlated with the actual values (r = 0.70), having an identical mean value of 117 +/- 15.3 and an unbiased regression relation (y = 0.000 + 1.000x). As standard measurements of the GFR are very time consuming and expensive, the use of the simple equation GFR1 = 218.1 - 0.916 x Age - 0.635 x Creatinine is recommended. The classification of GFR values into three ranges has also revealed the nonlinear characteristics of GFR in relation to other demographic variables: age and creatinine are the dominant variables in the middle GFR range, while the body-mass index and urea are dominant in the high and low ranges, respectively. PMID- 9216072 TI - Hearing in blind subterranean Zambian mole-rats (Cryptomys sp.): collective behavioural audiogram in a highly social rodent. AB - Thresholds for pure tone detection were examined in the common mole-rat, Cryptomys sp. (Bath-yergidae, Rodentia) using a positive reinforcement procedure. To bypass the problems connected with testing isolated individuals of this extremely social species, a collective behavioural audiogram was determined for a family group of seven mole-rats. Within the tested frequency range of 225 to 18 kHz, the lowest thresholds (as low as 7.5 dB SPL, on average 24 dB SPL) were found at 800 Hz, the upper limit of hearing (at the level of 60 dB SPL) was at 18 kHz. The behavioural audiogram combines the results of previous studies on hearing in this species. It resembles the distortion threshold curve but differs from neurophysiological data as far as the high frequency cutoff is concerned. On the other hand, the region of the best hearing sensitivity is narrow in behavioural audiogram and neurophysiological curves but rather broad in the distortion threshold curve. In general, the behavioural audiogram of Cryptomys is in many aspects comparable with the available audiograms of other subterranean rodents. PMID- 9216073 TI - Protein synthesis under conditions of anoxia and changing workload in ventricle strips from turtle heart. AB - An earlier study determined that protein synthesis in isolated perfused turtle (Trachemys [= Pseudemys] scripta elegans) hearts was three-fold lower under conditions of anoxia than under conditions of normoxia. However, the earlier study did not attempt to define the role of work in the isolated perfused preparation. In this study, the effects of varying workload, as defined by changing frequency of contraction, and anoxia on protein synthesis were examined. The ventricle strip preparation allows for comparison of multiple strips from a single heart, which aids in eliminating the variability found between individuals chosen from wild populations. Ventricle strips forced to contract at 24 contractions.min-1 under anoxic conditions failed more rapidly than strips forced to contract at 24 contractions.min-1 under normoxic conditions. Protein synthesis decreased by 32% when compared to normoxic controls. When stimulation was terminated after 2 hr of contraction, the rate of protein synthesis in strips under anoxic conditions was similar to that in strips under normoxic conditions. Also, returning strips to normoxic conditions after 2 hr of anoxia restored protein synthesis to the level of the normoxic controls. A significant correlation between pacing rate and protein synthesis was found under normoxic conditions but not under anoxic conditions when strips were paced at 12, 18 and 24 contractions.min-1. Protein synthesis increased by 30% at the 18 contractions.min-1 frequency and 45% at the 24 contractions.min-1 frequency over the rate at 12 contractions.min-1 frequency. Force-frequency studies revealed that under normoxic conditions force generation did not change until above 24 contractions.min-1, but under anoxic conditions there was a significant negative inotropic effect (20% decrease in force) at 24 contractions.min-1 and fell to 50% of initial at 36 contractions.min-1. These studies indicate that, in the turtle heart, anoxia per se is not the only determinant of protein synthesis but rather that work plays an important role in protein synthesis, as in the mammalian heart. PMID- 9216074 TI - Mitochondrial ammonia metabolism and the proton-neutral theory of hepatic ammonia detoxication. PMID- 9216075 TI - Isolation of histones and related chromatin structures from spermatozoa nuclei of a dasyurid marsupial, Sminthopsis crassicaudata. AB - The spermatozoa of a dasyurid marsupial, Sminthopsis crassicaudata, have two distinct nuclear regions: uniformly electron-dense chromatin (C1) in the interior and fissured chromatin (C2) at the periphery. To investigate whether the differences in morphology are due to incorporation of different packaging proteins, spermatozoa nuclear proteins were characterised by acetic acid-urea polyacrylamide gel electrophoresis (PAGE) and fractionated by reverse-phase high pressure liquid chromatography (HPLC). The main protein component was protamine I, but a complete histone complement (H1, H2A, H2B, H3, and H4) was also detected. Immunocytochemistry showed localisation of H4, H2B, and H2A histones to the periphery of the nuclei, a region that corresponded to the C2 chromatin. The fissures in the chromatin of this region disappeared following incubation with fish protamines, indicating that the nucleohistone C2 region may be incompletely condensed relative to nucleoprotamines. This observation is consistent with the view that 60% of phosphodiester charges remain negative in nucleohistone DNA, whereas all DNA charges are neutralised in highly compact nucleoprotamines. Treatment of spermatozoa with micrococcal nuclease showed that the C1 chromatin was resistant to digestion, whereas the C2 region was cleaved into 30- to 38-nm agglomerates and 11-nm nucleosomal-size structures. Thus, this study demonstrates that spermatozoa nuclei of this marsupial species contain peripherally localised histones, and the nucleohistone chromatin accounts for the different morphology of the C2 region compared with the rest of the nucleus. PMID- 9216076 TI - Suppression, accessibility of death-related thoughts, and cultural worldview defense: exploring the psychodynamics of terror management. AB - Previous research has shown that after a mortality-salience (MS) treatment, death thought accessibility and worldview defense are initially low and then increase after a delay, suggesting that a person's initial response to conscious thoughts of mortality is to actively suppress death thoughts. If so, then high cognitive load, by disrupting suppression efforts, should lead to immediate increases in death thought accessibility and cultural worldview defense. Studies 1 and 2 supported this reasoning. Specifically, Study 1 replicated the delayed increase in death accessibility after MS among low cognitive load participants but showed a reversed pattern among participants under high cognitive load. Study 2 showed that, unlike low cognitive load participants, high cognitive load participants exhibited immediate increase in pro-American bias after MS. Study 3 demonstrated that worldview defense in response to MS reduces the delayed increase in death accessibility. Implications of these findings for understanding both terror management processes and psychological defense in general are discussed. PMID- 9216077 TI - Lay dispositionism and implicit theories of personality. AB - Lay dispositionism refers to lay people's tendency to use traits as the basic unit of analysis in social perception (L. Ross & R. E. Nisbett, 1991). Five studies explored the relation between the practices indicative of lay dispositionism and people's implicit theories about the nature of personal attributes. As predicted, compared with those who believed that personal attributes are malleable (incremental theorists), those who believed in fixed traits (entity theorists) used traits or trait-relevant information to make stronger future behavioral predictions (Studies 1 and 2) and made stronger trait inferences from behavior (Study 3). Moreover, the relation between implicit theories and lay dispositionism was found in both the United States (a more individualistic culture) and Hong Kong (a more collectivistic culture), suggesting this relation to be generalizable across cultures (Study 4). Finally, an experiment in which implicit theories were manipulated provided preliminary evidence for the possible causal role of implicit theories in lay dispositionism (Study 5). PMID- 9216078 TI - Repressive coping: distraction using pleasant thoughts and memories. AB - To avoid exposure to unpleasant or unwanted emotional material, some people may distract themselves by summoning up pleasant thoughts such as happy memories. Manipulation of negative affect might therefore result in heightened accessibility of pleasant thoughts and memories, contrary to hypotheses of mood congruent recall. In Experiment 1, repressors were faster to recall happy memories after watching an unpleasant film than after watching a neutral film. Nonrepressors showed the opposite effect (i.e., mood-congruent memory). In Experiment 2, after an unpleasant film, repressors were faster to recall a happy memory than to recall a sad memory. In Experiment 3, repressors spontaneously generated pleasant thoughts after watching an unpleasant film, whereas nonrepressors did not. Thus, repressors apparently cope with exposure to negative affective material by accessing pleasant thoughts. Results are discussed in terms of cognitive defenses against emotional distress and the associative structure of repression. PMID- 9216079 TI - Cognitive and physiological antecedents of threat and challenge appraisal. AB - Cognitive appraisal theories of stress and emotion propose that cognitive appraisals precede physiological responses, whereas peripheralist theories propose that physiological arousal precedes cognitive processes. Three studies examined this issue regarding threat and challenge responses to potential stress. Study 1 supported cognitive appraisal theory by demonstrating that threat and challenge cognitive appraisals and physiological responses could be elicited experimentally by manipulating instructional set. Studies 2 and 3, in contrast, found that manipulations of physiological response patterns consistent with challenge and threat did not result in corresponding changes in cognitive appraisal. Appraisals in Study 3, however, were related to subjective pain independent of the physiological manipulation. These studies suggest a central role for cognitive appraisal processes in elicitation of threat and challenge responses to potentially stressful situations. PMID- 9216080 TI - Negative life events, marital interaction, and the longitudinal course of newlywed marriage. AB - Life events and problem-solving behavior were examined relative to longitudinal change in depressive symptoms and marital adjustment over 18 months in 60 newlywed couples. Spouses' problem-solving behavior moderated, but did not mediate, the relationship between life events and adjustment. Some behaviors contributed to spouses being more resilient to life events, and some behaviors made spouses more vulnerable. In particular, wives' anger facilitated their adjustment to major and interpersonal events such that their depressive symptoms declined and their marital satisfaction increased. Husbands' humor contributed to marital instability when spouses reported more major events. The results further specify the vulnerability-stress-adaptation model of marriage and expand on the role of behavior in marriage. PMID- 9216081 TI - Evidence for genetic influence on both cross-situation and situation-specific components of behavior. AB - An assumption often made in the study of personality and in social psychology is that methods variance and situation-specific effects, as key components of measured behavioral variance, are environmental effects. The results of the present research refute that assumption. Nine measures-3 aspects of temperament measured in each of 3 ways-were obtained at age 24 months for twin sibships participating in the Louisville Twin Study. This report describes a new model that captures the unique information potentially available in such data, by combining multitrait-multimethod and twin-family analytic designs. The results indicated significant genetic influence on methods-situations components of variance along with genetic influence on traits. The findings support heuristics that include both situation-specific patterns of behavior and cross-situational consistencies. PMID- 9216082 TI - Differential roles of neuroticism, extraversion, and event desirability for mood in daily life: an integrative model of top-down and bottom-up influences. AB - Top-down and bottom-up approaches were combined to assess the relative impact of extraversion, neuroticism, and daily events on daily mood. Ninety-six community residing men completed diaries for 8 consecutive nights. Extraversion predicted positive mood, whereas neuroticism predicted positive and negative mood. Undesirable events predicted negative mood and, more modestly, positive mood. Desirable events predicted positive mood. Negative dispositional and situational factors play a larger role in daily positive affect than positive factors do in daily negative affect. PMID- 9216083 TI - Avoidance achievement motivation: a personal goals analysis. AB - The present research investigated one antecedent and various consequences of pursuing avoidance personal achievement goals over the course of a semester. The authors assessed participants' achievement-relevant goals using the newly devised Achievement Goals Questionnaire. The motive to avoid failure, assessed with self report and projective measures, was established as an antecedent of avoidance goal pursuit. Avoidance regulation was shown to have deleterious consequences for a host of achievement-relevant and general well-being outcomes at the end of the semester, including longitudinal change in subjective well-being. Perceived competence was validated as a mediator of the direct relationships observed. The results highlight the need to attend to avoidance, as well as approach, forms of self-regulation and the need to consider both motive disposition and goal constructs in accounting for competence-relevant behavior. PMID- 9216084 TI - The crystal structure of a Fab fragment to the melanoma-associated GD2 ganglioside. AB - The GD2 ganglioside is a cell-surface component that appears on the surface of metastatic melanoma cells and is a marker for the progression of the disease. The ME36.1 monoclonal antibody binds to the GD2 ganglioside and has shown potential as a therapeutic antibody. ME36.1 is a possible alternative therapy to radiation, which is often ineffective in late-stage melanoma. The crystal structure of the Fab fragment of ME36.1 has been determined using molecular replacement and refined to an R factor of 20.4% at 2.8 A resolution. The model has good geometry with root-mean-square deviations of 0.008 A from ideal bond lengths and 1.7 degrees from ideal bond angles. The crystal structure of the ME36.1 Fab shows that its complementarity determining region forms a groove-shaped binding site rather than the pocket-type observed in other sugar binding Fabs. Molecular modeling has placed a four-residue sugar, representative of GD2, in the antigen binding site. The GD2 sugar moiety is stabilized by a network of hydrogen bonds that define the specificity of ME36.1 toward its antigen. PMID- 9216085 TI - Polymorphic fibrillar assembly of human amylin. AB - Human amylin forms fibrillar amyloid between pancreatic islet cells in patients with non-insulin-dependent (type 2) diabetes mellitus. Fibrillar assemblies also form in vitro in aqueous solutions of synthetic human amylin. We now report on the structural polymorphism of these fibrils. The thinnest fibril, referred to as the protofibril, has an apparent width of 5 nm but is only rarely observed by itself. These protofibrils spontaneously assemble into higher order fibrillar structures with distinct morphologies. Prominent among these is an 8-nm fibril with a distinct 25-nm axial crossover repeat which is formed by left-handed coiling of two 5-nm protofibrils. Coiling of more than two 5-nm protofibrils results in cable-like structures of variable width depending on the number of protofibrils involved. Lateral (side-by-side) assembly of 5-nm protofibrils is also observed and produces ribbons which may contain two, three, four, or more protofibrils and occasionally large single-layered sheets. The mass-per-length (MPL) of the 5-nm protofibril is 10 kDa/nm. This has been established in two ways: first, the 8-nm fibril, which is formed by coiling two 5-nm protofibrils around each other, has an MPL of 20 kDa/nm. Second, higher order fibrils differ by increments of 10 kDa/nm. Hence, about 2.6 human amylin molecules (3904 Da) are packed in 1 nm of protofibril length. Similarities exist between amylin fibrils and those formed from other amyloid proteins, suggesting that the in vitro assembly of synthetic protein may serve as a useful model system in advancing our understanding of amyloid formation in disease. PMID- 9216086 TI - Crystallization and preliminary X-ray analysis of the single-headed and double headed motor protein kinesin. AB - Crystals of the single-headed and double-headed kinesin motor domains of Rattus norvegicus have been grown by vapor diffusion using ammonium sulfate as the precipitant. Both crystal systems belong to the orthorhombic space group P2(1)2(1)2(1). Double-headed kinesin crystallized with unit cell constants of a = 72.2 A, b = 91.9 A, and c = 141.7 A, and so far the best crystals diffracted to a maximum resolution of 2.7 A. Using ammonium sulfate single-headed kinesin crystallized in two different crystal forms with cell constants of a = 73.1 A, b = 73.2 A, c = 84.0 A and a = 73.4 A, b = 74.1 A, c = 74.7 A, respectively. They were found to diffract to 2.1 A resolution. Crystals of monomeric kinesin were also obtained with lithium sulfate as precipitant. They have cell constants of a = 71.6 A, b = 73.7 A, and c = 74.1 A and diffract up to 1.7 A resolution. PMID- 9216088 TI - The toxicity of the Alzheimer's beta-amyloid peptide correlates with a distinct fiber morphology. AB - In an attempt to elucidate the relationship among aggregation properties, fiber morphology, and cellular toxicity several beta-amyloid peptides (A beta) were prepared according to a standardized procedure. Peptides either carried mutations inside the membrane anchor segment around amino acid position 35 or their carboxy terminus was shortened from 42 to 41, 40, or 39 amino acids. The time-dependent self-assembly of monomeric A beta into fibers was simultaneously monitored by electron microscopy, circular dichroism spectroscopy, analytical ultracentrifugation, and A beta-mediated cellular toxicity using the reduction of 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) to measure cell viability. The transition of A beta monomers into fibers was analyzed by more than 600 electron micrographs. Distinct morphological changes from seed-like structures to immature and mature fibers were observed. Seeds were of spherical appearance. Immature fibers were typically elongated structures with a rough surface and with varying thickness depending on the A beta sequence. Mature fibers were characterized by a periodic variation of their thickness along the fiber axis. The proportion of these different structures and the total amount of aggregated A beta was amino acid sequence-dependent. Wild-type A beta 1-42 and its oxidized derivative carrying a methionine sulfoxide residue at position 35 showed the highest rate of fiber formation and exerted toxic activity in the MTT assay at very low nanomolar concentrations. The fibers formed by these two peptides were predominantly of the mature type. In contrast, carboxyl-terminus truncated peptides A beta 1-41, A beta 1-40, and A beta 1-39 or most A beta 1-42 derivatives mutated around amino acid position 35 showed a reduced aggregation rate, the immature fibers predominated, and the toxicity was orders of magnitude lower. Thus, a correlation can be drawn among the chemical structure, aggregation properties, fiber morphology, and cellular toxicity. PMID- 9216087 TI - Visualization of prosomes (MCP-proteasomes), intermediate filament and actin networks by "instantaneous fixation" preserving the cytoskeleton. AB - A new "instantaneous" fixation/extraction procedure, yielding good preservation of intermediate filaments (IFs) and actin filaments when applied at 37 degrees C, has been explored to reexamine the relationships of the prosomes to the cytoskeleton. Prosomes are protein complexes of variable subunit composition, including occasionally a small RNA, which were originally observed as trans acting factors in untranslated mRNPs. Constituting also the proteolytic core of the 26S proteasomes, they are also called "multicatalytic proteinase (MCP) complexes" or "20S-Proteasomes." In Triton X-100-extracted epithelial, fibroblastic, and muscle cells, prosome particles were found associated primarily with the IFs (Olink-Coux et al., 1994). Application of "instantaneous fixation" has now led to the new observation that a major fraction of prosome particles, composed of specific sets of subunits, is distributed in variable proportions between the IFs and the microfilament/ stress fiber system in PtK1 epithelial cells and human fibroblasts. Electron microscopy using gold-labeled antibodies confirms this dual localization on classical whole mounts and on cells exposed to instantaneous fixation. In contrast to the resistance of the prosome-IF association, a variable fraction of the prosome particles is released from the actin cytoskeleton by Triton X-100 when applied prior to fixation. Moreover, in vitro copolymerization of prosomes with G-actin made it possible to observe "ladder-like" filamentous structures in the electron microscope, in which the prosome particles, like the "rungs of a ladder," laterally crosslink two or more actin filaments in a regular pattern. These results demonstrate that prosomes are bound in the cell not only to IFs but also to the actin cytoskeleton and, furthermore, not only within large M(r) complexes (possibly mRNPs and/or 26S proteasomes), but also directly, as individual prosome particles. PMID- 9216090 TI - Measurement of gastric emptying time by real-time ultrasonography in patients with Crohn's disease. AB - In order to gain some insight into the possible influence of gastric emptying on gallbladder hypomotility in patients with Crohn's disease, the gastric emptying time (GET) was measured by means of ultrasonography in 10 healthy controls and 10 patients with Crohn's disease. No significant difference was observed between both mean values for GET studies (GET: controls, 165.0 min +/- 12.8; Crohn, 142.0 min +/- 11.5; p = 0.208) after ingestion of a liquid meal. Thus, the gallbladder hypomotility described in patients with Crohn's disease, after a liquid fatty meal stimulus, can not be explained by prolonged gastric emptying time. PMID- 9216089 TI - Reproducibility of the ultrasound method for measurement of gallbladder volume. AB - Gallbladder motility has largely been studied in recent years. Since the ultrasonographic method can be used in gallbladder emptying studies, we investigated the reproducibility of the ultrasound method for measurement of gallbladder volume. The ultrasonographic method was highly reproducible (r = 0.97) and, due to its safeness and lack of use of radioactive agents, it is attractive option for gallbladder motility studies in conditions associated with increased frequency of cholelithiasis. PMID- 9216091 TI - Diarrhea and malabsorption in primary humoral immunodeficiency. AB - Patients with Humoral immunodeficiency syndromes frequently present recurrent infections, mainly of the digestive and respiratory tracts. This study carried out a clinical and laboratorial evaluation in 15 humoral immunodeficiency patients presenting chronic gastrointestinal symptoms. Out results emphasize the relevance of immunodeficiency syndromes in the differential diagnosis of chronic diarrhea. PMID- 9216093 TI - Pathogenesis of chagasic myocarditis: an experimental study. AB - With the propose to study the pathogenesis of chagasic myocarditis, white mice were inoculated with the Y strain of Trypanosoma cruzi. After confirmation of parasitemia, the animals were sacrificed: a) in the acute stage (first 30 days), 5 mice/day: b) in the subacute stage (days 30-90), 5 mice/week: c) in the chronic stage the remaining survivors, 365 days after inoculation. Parasitemia appeared on the sixth day and increased gradually until the maximum was reached in the third or fourth week, then decreasing until the thirtieth day, disappearing from the circulation after the 87th day. Parasitism occurred with parasitemia however, without a quantitative relationship. Starting from the sixth post-inoculation day the muscle fibers demonstrated parasites, with the presence of compact or loose amastigote nests. The nests, bursted during the evolution so that their contents (cytokine or interleukin?) could extravasate. While the nest was integral, there was no acute inflammatory reaction. During the chronic stage empty spaces remained and were occupied by the chronic inflammatory process: proliferation of fibroblasts. granulomas and residual fibrosis. The inflammatory edema replaced the interstitium. After rupture of the amastigote nests they were initially invaded by the acute inflammatory process and then by the chronic one with deposition of a fibrin network, proliferation of collagen, mastocytosis and terminating with fibrosis. It was not uncommon to find amastigote nests in various stages during the chronic phase in the myocardial fibers, characterizing the cyclical process of the parasitosis. The blood vessels were also, invaded by parasites. Causing thrombosis and necrotizing arteritis. The autonomous nervous system also was involved, with the presence of amastigote nests, ganglionitis, plexulitis with inflammation in scattered areas. PMID- 9216092 TI - Five day and ten day triple therapy (amoxicillin, furazolidone and metronidazole) in the treatment of duodenal ulcer. AB - This investigation aimed to compare bacterial eradication and healing in patients with active duodenal ulcer treated with a combination of furazolidone 600 mg/day and metronidazole 750 mg/day and amoxicillin 1.5 and g/day for 5 (TT5) or 10 (TT10) days. Fifty four (TT5 = 28 and TT10 = 26) patients were included in the study. Ulcer healing was observed in 77.8% of TT5 Group and in 75% of TT10 Group at week 4. H pylori eradication was observed in 51.9% and 65% respectively (p > 0.05). When all patients were grouped, a significantly healing rate was observed in those eradicated as compared to those not eradicated (p = 0.03). We concluded that extending the treatment to 10 days did not significantly influence the results of ulcer healing and eradication of Helicobacter pylori. PMID- 9216094 TI - Secretory, endoscopic and histopathologic changes and prevalence of Helicobacter pylori in the gastroduodenal mucosa in patients with chronic alcoholic pancreatitis. AB - Aiming at establishing the prevalence of peptic ulcer in chronic alcoholic pancreatitis and an eventual correlation with gastric acid secretion and endoscopic and histopathologic alterations as well as the presence of Helicobacter pylori in the gastroduodenal mucosa, thirty patients with chronic alcoholic pancreatitis (Group I) and ten control subjects (Group II) were prospectively studied. After upper gastrointestinal endoscopy was performed. Group I was subdivided according to the lack (Subgroup Ia) or a presence (Subgroup Ib) of peptic ulcer. The prevalence of peptic ulcer in these patients was 23.33% clearly higher than that reported in the general population. Baseline and stimulated acid secretion as well as baseline gastrinemia among the subgroups and groups were similar. There was no statistically significant difference in the other parameters evaluated. Due to the increased prevalence of asymptomatic peptic ulcer in patients with chronic alcoholic pancreatitis. Upper gastrointestinal endoscopy is suggested as a diagnosis routine and follow-up of this group of patients. PMID- 9216096 TI - Changes in capillary permeability during severe experimental acute pancreatitis in rats. AB - Abnormalities of microcirculation are thought to have an important role in the pathogenesis of severe acute pancreatitis (SAP) and in the associated multiple organic failure. We studied changes in capillary permeability during experimental SAP in rats. Necrotizing acute pancreatitis was induced by retrograde injection of sodium taurocholate in pancreatic ducts in rats (n = 30). The animals were distributed in three groups in which the spaces of compartmental distribution as well as the tissue distribution of labeled tracers were evaluated with the use of erythrocytes and albumin tagged with radioactive chromium, apart from determining the volume of total body water. All the studies were carried out four hours after the induction of acute pancreatitis and the administration of radioactive tracers. PMID- 9216095 TI - Clearance of hepatitis C viral RNA in cirrhotic patients with antiviral therapy. AB - Interferon is indicated in chronic infection by hepatitis C virus (HCV), however, cirrhosis has been reported as a bad response factor to the therapy. Fifteen cirrhotic patients with HCV, undergoing treatment with recombinant interferon alpha, ribavirin and/or ursodeoxycholic acid were studied. They were followed-up and evaluated with dosages of alanine aminotransferase and HCV RNA investigation by PCR technique. Of the 15 cirrhotic patients, seven were negative for HCV RNA after antiviral treatment, however ALT was normal in only three of them. Of the eight patients who were not negative, two had normal ALT. Biochemical-virological discrepancy in the follow-up of the patients after antiviral treatment observed in this study has also been reported by other authors. These reports show that the criteria for response to the treatment is to be established. PMID- 9216097 TI - Hyperbaric oxygen: a new alternative in the treatment of perianal Crohn's disease. PMID- 9216098 TI - Primary extranodal non-Hodgkin lymphoma of the extrahepatic bile duct mimmicking Klatskin tumor. AB - This report describes one case of primary non-Hodgkin lymphoma of the extrahepatic biliary tree. The main symptom was obstructive jaundice. Cholangiography demonstrated stricture of the bile duct which resembled the appearance of cholangiocarcinoma. The surgical approach allowed complete ressection. The histopathological analyses showed a centrocitic-centroblastic follicular non-Hodgkin lymphoma. She underwent chemotherapy, developed severe bone marrow hypoplasia, but 48 months after surgery, the patient is doing well. PMID- 9216100 TI - Cholesterol gallstone formation: supersaturation, nucleation and gallbladder motility. PMID- 9216099 TI - Squamous cell carcinoma of the pancreas with gastric metastasis. Case report. AB - The squamous cell carcinoma of the pancreas is an uncommon form of pancreatic cancer, with a frequency in the range of 0.5-3.5%. A rare case of a primary squamous cell pancreatic carcinoma, with gastric invasion and upper digestive bleeding, requiring surgical control is reported. The surgical technique to treat the bleeding is also detailed. PMID- 9216101 TI - Viral hepatitis prophylaxis. AB - Acute viral hepatitis is the most usual cause of jaundice and acute liver failure, whereas chronic viral hepatitis is the major cause of liver cirrhosis and hepatocellular carcinoma. Taking into the consideration the morbidity and mortality of such lesions, their prophylaxis is a mandatory procedure. In this review we discuss the general measures and the active and passive immunoprophylaxis against hepatitis A. B and Yellow fever, and the general management of hepatitis C. D. and E virus infection. PMID- 9216102 TI - Interferon in viral liver diseases: pharmacological aspects. AB - In 1957, two English investigators, Alick Isaacs and Jean Lindenmann made a very important discovery. They observed that when a virus species colonized cells in animals, this invasion interfered with the ability of other viruses, without any association with the former, to produce simultaneous infection. Both investigators observed that the substance responsible for the inhibition was secreted by the infected cells and called it interferon. They also observed that this protein did not directly interact with the virus, but induced the infected cells and surrounding cells to produce other proteins, which were in turn able to block the multiplication of the invading virus. PMID- 9216103 TI - Mercury contamination and health risk in the Brazilian Amazon. An ethical dilemma. PMID- 9216104 TI - Concomitant rotavirus serotypes 1 and 4 infections in a diarrhoeic child from Belem, Brazil. AB - Concomitant serotypes 1 and 4 infections were detected in a 15-month old female child with community-acquired diarrhoea which lasted 7 days and coursed with moderate dehydration. The evidence for dual rotavirus infection was offered by the following findings: a) enzyme-linked immunosorbent assay (ELISA) positive reactions to both 1 and 4 serotypes; and b) extra-migrating bands at electrophoresis of RNA in polyacrylamide gel (PAGE). These results suggest that children living under poor sanitation conditions are heavily exposed to rotavirus infections; in addition, the co-circulation of different serotypes in the same setting sustains the current concept that a rotavirus vaccine should be multivalent, in order to protect children against the four epidemiologically important rotavirus G serotypes. PMID- 9216105 TI - Active replication of hepatitis B virus (HBV) in HIV type 1 and in HIV type 2 infected patients. AB - To evaluate the effect of concurrent infection by HIV on HBV infection or immunity, we have studied a group of 66 HIV1+ symptomatic Caucasian patients and another of 38 African HIV2+ asymptomatic individuals, concerning their HBV status: serological markers of infection and presence of HBV-DNA in serum, the last taken as sign of hepatitis B virus active replication, were monitored. HIV+ groups were compared with seronegative controls, adequately matched for age, sex and ethnological background. HBV DNA was found in 7.6% of HIV1+ Caucasian patients and 3.2% of seronegative controls; in African HIV2+ individuals 2.6% were also HBV DNA+, a percentage close to that found in HIV2 seronegative controls (2.9%). No correlation was found between HIV infection and HBV active replication. Immunodepression that follows HIV infection over time may be compatible with a degree of T cell function capable of avoiding reinfection with or reactivation of HBV, even in symptomatic stages of acquired immunodeficiency syndrome. Our findings are relevant to the choice of preventive strategies in populations at risk for HIV and HBV infection. PMID- 9216106 TI - Immunoperoxidase for the detection of antibodies in cerebrospinal fluid in neurocysticercosis: use of Cysticercus cellulosae and Cysticercus longicollis particles fixed on microscopy slides. AB - The ORF strain of Cysticercus longicollis represents an important model for the study of heterologous antigens in the immunodiagnosis of neurocysticcreosis (NC). The immunoperoxidase (IP) technique was standardized using a particulate antigen suspension of Cysticercus longicollis (Cl) and Cysitcercus cellulosae (Cc). Cerebrospinal fluid (CSF) samples were incubated on the antigen fixed to microscopy slides; the conjugate employed was anti-IgG-peroxidase and the enzymatic reaction was started by covering the slides with chromogen solution (diaminobenzidine/H2O2). After washing with distilled water, the slide was stained with 2% malachite green in water. Of the CSF samples from 21 patients with NC, 19 (90.5%) were positive, whereas the 8 CSF samples from the control group (100%) were negative. The results of the [P-C] test applied to 127 CSF samples from patients with suspected NC showed 28.3% reactivity as opposed to 29.1% for the IP-Cc test. The agreement index for the IP test (Cl x Cc) was 94.2%, with no significant difference between the two antigens. PMID- 9216107 TI - Ascariasis in the subdistrict of Cavacos, municipality of alterosa (MG), Brazil: effect of mass treatment with albendazole on the intensity of infection. AB - The clinical and epidemiologic aspects of infection with Ascaris lumbricoides were studied in a random stratified sample of the population of the subdistrict of Cavacos, municipality of Alterosa (Minas Gerais, Brazil). The effect of mass treatment with a single dose of albendazole on the prevalence and intensity of infection was also studied six months later in the same population. During the first phase of the study, a questionnaire was applied to 248 individuals to obtain information about the socioeconomic, sanitary and clinical conditions of the population surveyed. A total of 230 fecal samples were also examined by the Kato-Katz technique in order to determine the intensity of A. lumbricoides infection. Two hundred and two individuals were simultaneously submitted to blood counts and 70 children aged 12 years or less were evaluated for nutritional status. The presence of A. lumbricoides and other helminth eggs was also determined in 22 soil samples collected in the urban zone of Cavacos. Infection with enteroparasitic helminths was detected in 29.1% of the sample, with a predominance of A. lumbricoides (23.9%). Parasitism and/or intensity of A. lumbricoides infection were significantly correlated with age range (15 years or less), social class, sanitary and living conditions (water, sewage and domiciliary area per person), and presence of abdominal pain. However, these parameters were not correlated with nutritional status or hematocrit levels. During the second phase of the study, a slight but not statistically significant decrease in the prevalence of A. lumbricoides infection was detected after treatment with albendazole. However, an important and significant reduction in the amount of A. lumbricoides eggs eliminated through the feces was detected, indicating that the intensity of A. lumbricoides infection was lower in all the age ranges of the Cavacos population, especially among younger individuals, even six months after administration of the anthelminthic agent. PMID- 9216108 TI - New records of Histoplasma capsulatum from wild animals in the Brazilian Amazon. AB - Twenty-eight isolates of Histoplasma capsulatum were obtained from eight species of forest mammals from the States of Amazonas, Para and Rondonia in the Amazon Region of Brazil. Primary isolates were obtained by inoculating triturated liver and spleen tissue intradermally and intraperitoneally in hamsters. Mycological diagnosis in hamsters presenting lesions was confirmed by histopathology and culture on Sabouraud dextrose-agar. Infected hamsters developed signs of disease within two to nine months; all had disseminated visceral lesions and most also had skin lesions at the sites of inoculation. None of the hamsters inoculated with skin macerates of the original hosts developed histoplasmosis, and histopathological examination of the viscera of the wild hosts failed to reveal H. capsulatum. Prevalence of infection was considerably higher in females than in males both for the opossum Didelphis marsupialis and for total wild animals (479) examined. It is proposed that canopy-dwelling mammals may acquire the infection from conidia borne on convective currents in hollow trees with openings at ground level. PMID- 9216110 TI - Tumor-like lesion due to Chagas' disease in a patient with lymphocytic leukemia. AB - A 73 year-old white male, living in the interior of the state of Mato Grosso do Sul, in central Brazil, after an initial diagnosis of sinusitis was transferred to the neurology service with a 3-day evolution of intracranial hypertension. Exams showed lymphocytic leukemia and a tumor-like lesion, either an expanding inflammatory process such as an abscess or a neoplasm. Treatment with Ceftriaxone and Decadron was started and intracranial hypertension was controlled. Methotrexate was injected on the occasion of the next puncture considering a possible leukemia infiltration. Flagellate forms of T. cruzi were observed in the CSF and treatment with Benznidazole was started. After 4 days the CSF presented fractionated forms of trypomastigotes. The protein level was 27%. Signs of intracranial hypertension ceased. Tomography and magnetic resonance images showed an important reduction of the tumor-like lesion. The clinical condition of the patient improved. PMID- 9216109 TI - Studies on prevalence of Strongyloides infection in Holambra and Maceio, Brazil, by the agar plate faecal culture method. AB - Prevalence of Strongyloides stercoralis infection in three areas of Brazil was surveyed by a recently developed faecal culture method (an agar plate culture). The Strongyloides infection was confirmed in 11.3% of 432 subjects examined. The diagnostic efficacy of the agar plate culture was as high as 93.9% compared to only 28.5% and 26.5% by the Harada-Mori filter paper culture and faecal concentration methods, when faecal samples were examined simultaneously by these three methods. Among the 49 positive samples, about 60% were confirmed to be positive only by the agar plate culture. These results indicate that the agar plate culture is a sensitive new tool for the correct diagnosis of chronic Strongyloides infection. PMID- 9216111 TI - Infection by Trypanosoma cruzi in mammals in Yucatan, Mexico: a serological and parasitological study. AB - In order to determine Trypanosoma cruzi infection among mammals in Yucatan, Mexico, 372 animals, both wild and synanthropic including carnivores, marsupials and rodents were studied. Serological studies by indirect haemagglutination (IHA) were carried out to detect antibodies to T. cruzi and a parasitological study was also performed (blood smear and histopathology). Of all the animals tested 18.54% were serologically positive, with a significantly higher frequency among the wild ones (33.33%) compared to the synanthropic ones (17.79%). To determine T. cruzi in positive animals, blood was inoculated into a white mouse (webster type) to prove myocardium colonization. The serological and parasitological positivity of these animals, as well as their behavior in the environment, taken together with the socioeconomic and cultural characteristics of the population, suggest that in Yucatan, Mexico, Canis familiaris, Didelphis marsupialis and Rattus rattus act as a link with the wild cycle. PMID- 9216112 TI - Effects of ivermectin on Culex quinquefasciatus larvae. AB - The effects of ivermectin, a semi-synthetic drug widely used for treatment of livestock parasite diseases, were observed on Culex quinquefasciatus larvae. Toxic effects and mortality evaluation were carried out after 5, 15, 30 and 60 minutes of exposure to 1, 5 or 10 ppm of ivermectin solutions. Observations were made 24 and 48 hours after the beginning of the experiment, and loss of mobility, progressive paralysis and high mortality of larvae were recorded. The observed effects of ivermectin on the mosquito larvae is probably correlated with chloride channel activation on cell membranes. PMID- 9216113 TI - A single method to stain Malassezia furfur and Corynebacterium minutissimum in scales. AB - The scales are collected by pressing small pieces of scotch tape (about 4 cm length and 2 cm width) onto the lesions and following withdrawal the furfuraceous scales will remain on the glue side. These pieces are then immersed for some minutes in lactophenol-cotton blue stain. Following absorption of the stain the scales are washed in current water to remove the excess of blue stain, dried with filter paper, dehydrated via passage in two bottles containing absolute alcohol and then placed in xylene in a centrifugation tube. The xylene dissolves the scotch tape glue and the scales fall free in the tube. After centrifugation and decantation the scales concentrated on the bottom of the tube are collected with a platinum-loop, placed in Canada balsam on a microscopy slide and closed with a cover slip. The preparations are then ready to be submitted to microscopic examination. Other stains may also be used instead of lactophenol-cotton blue. This method is simple, easily performed, and offers good conditions to study these fungi as well as being useful for the diagnosis of the diseases that they cause. PMID- 9216114 TI - Schistosoma mansoni: exacerbation of inflammatory granulomatous response in mice chronically infected and submitted to reinfection. PMID- 9216115 TI - Schistosoma mansoni Sambon, 1907: effects of dilation and constricting anesthetics drugs on adult worms localization in Swiss mice. PMID- 9216116 TI - Prevalence of hepatitis C antibodies among health care workers at high risk for blood exposure. PMID- 9216117 TI - Efficacy and safety of fenofibrate or gemfibrozil on serum lipid profiles in Chinese patients with type IIb hyperlipidemia: a single-blind, randomized, and cross-over study. AB - BACKGROUND: The purpose of the study was to evaluate the effects and safety of fenofibrate or gemfibrozil on plasma lipid profiles in Chinese patients with type IIb hyperlipidemia. METHODS: Patients with previously diagnosed type IIb hyperlipidemia were initially evaluated in this single-blind, randomized, cross over study. Only those who had persistent hyperlipidemia after a two-month diet control period were included. The patients were randomized to either fenofibrate 100 mg tid or gemfibrozil 600 mg bid for three months, then shifted to alternate drug for another three months after a two-month washout period. During the whole 10-month period, a prudent diet was maintained. Each patient was followed up at Out-patient Clinic regularly for diet interview, compliance and possible adverse events. RESULTS: A total of 12 patients, 11 males and 1 female, completed the whole study. After a three-month treatment of fenofibrate, there was significant reduction in plasma total cholesterol (22.7%, p < 0.001), total triglycerides (43.2%, p < 0.001), and low-density lipoprotein (LDL)-cholesterol (25.7%, p = 0.001). Serum high density lipoprotein (HDL) cholesterol was also increased by 30.2% (p = 0.055). On the other hand, only total triglyceride was significantly reduced by 36.5% (p = 0.005) with gemfibrozil treatment. There was no significant change of serum total, HDL- and LDL-cholesterol after the three-month treatment of gemfibrozil. Besides, serum uric acid and alkaline phosphatase were significantly reduced with fenofibrate but not with gemfibrozil treatment. There was no obvious adverse event noted during each treatment period with either drug. CONCLUSIONS: The findings suggest that in Chinese patients with type IIb hyperlipidemia, both fenofibrate and gemfibrozil can safely and significantly reduce serum triglyceride. Fenofibrate, but not gemfibrozil, could also reduce serum total cholesterol, LDL and uric acid. The results were compatible with the findings of previous studies in western patients with hyperlipidemia. PMID- 9216118 TI - Comparison of risk factors for lacunar infarcts and other stroke subtypes. AB - BACKGROUND: Lacunar infarction (LI) is an ischemic stroke subtype with unique clinical, radiological and pathological features. Its relation to other stroke subtypes is unclear. To better understand the underlying pathological process of LI, we compared the risk factors of LI with those of other stroke subtypes. METHODS: During the study period (from January 1, 1990 to December 31, 1991), 240 consecutive patients with first-ever strokes admitted to the stroke unit of our hospital were enrolled to the study and were classified into one of the four stroke subtypes (52 with LI, 80 atherothrombotic infarcts, 38 cardiogenic embolism and 70 brain hemorrhage) based on their computed tomography (CT) and clinical features using the guideline developed by the National Institute of Neurological Disorders. Eighty outpatients of similar age who had either low back pain or cervical spondylosis were recruited from the clinics of Neurology to serve as non-stroke controls. Data collected included demographics, lifestyle, and other vascular risk factors. Detailed physical and neurological examination, blood biochemistry and Doppler ultrasound on cervical vessels were performed. RESULTS: Our investigations revealed that LI is a common stroke subtype accounting for 21% of all first-ever strokes in our hospital. Like ischemic stroke patients, those with LI were much more likely to have hypertension, diabetes, heart disease and carotid disease when compared with non-stroke controls. Patients with brain hemorrhage had less history of diabetes and lower levels of cholesterol than LI patients. CONCLUSIONS: LI patients seemed to share more risk factors with ischemic stroke patients than with brain hemorrhage patients. These shared risk factors suggest a possibly similar underlying pathological process between ischemic strokes and LI patients. Careful screening for those risk factors should be part of the mandatory clinical management for the prevention of LI. PMID- 9216119 TI - The clinical experience of gaseous retroperitoneoscopic and gasless retroperitoneoscopy-assisted unroofing of renal cyst. AB - BACKGROUND: The aim of this study is to compare the application of gaseous retroperitoneoscopic (GR) and gasless retroperitoneoscopy-assisted (GLRA) unroofing of renal cysts. METHODS: Fourteen patients with symptomatic simple renal cysts had undergone unroofing of the cyst with GR in seven cases and GLRA in seven others. Three trocars (10 mm, 10 mm and 5 mm) were inserted in the GR procedure. A 3 cm flank muscle-split incision was made and retroperitoneoscopy was performed through the same incision in the GLRA procedure. Then, the cyst was unroofed. RESULTS: The mean operative time was 104.3 minutes in the GR group and 52.1 minutes in the GLRA group, respectively (p = 0.001). The mean requirement of postoperative meperidine hydrochloride injection was 21.4 mg in the GR group and 71.4 mg in the GLRA group, respectively (p = 0.017). In the GR group, the mean postoperative stay was 3.7 days, and the time needed for return to normal activity was 7 days. In the GLRA group, the mean postoperative stay was 4.6 days, and the time needed for return to normal activity was 8 days. CONCLUSIONS: GR and GLRA techniques for unroofing of renal cysts are safe, effective and minimally invasive. GLRA is easy to perform and a more time-saving procedure when compared to GR, however, the patients of GLRA suffered more postoperative pain than after GR. GLRA is recommended in patients who had received retroperitoneal surgery or who have multiple renal cysts. PMID- 9216120 TI - Giant-cell tumor of bone around the knee. AB - BACKGROUND: In most of the reported series, almost half of the giant-cell tumors involved the knee region. The characteristics of the lesions were usually benign but often locally aggressive and easily recurrent neoplasms. Some surgeons performed intralesional excision combined with local adjunctive chemical coagulant, and bone grafting or methylmethacrylate cement packing. These methods could both decrease local recurrence and retain the function of joint. METHODS: From January, 1984 to December, 1994, a review was made of the results for eighteen patients who had been managed consecutively at the Veterans General Hospital-Taipei for giant-cell tumor of bone around the knee. Fourteen instances had occurred in the proximal tibia and four, in the distal femur. According to the classification of Campanacci, nine lesions were Stage II and nine, Stage III. Eleven patients had been managed with intralesional excision of the tumor with local adjunctive application; the other seven had en bloc resection and reconstructive procedures. RESULTS: All patients had been followed for a mean of fifty-six months (range 22 to 125 months). The overall recurrence rate was 11% (2/18). The intralesional excision had 18% (2/11) recurrence; there was no recurrence in the en bloc resection (0/7). The complication rate was 16% (4/18); 9% (1/11) for intralesional excision and 42% (3/7) for en bloc resection, respectively. The mean functional score was 28 points (range, 22 to 30) in the intralesional excision group and 21 points (range, 11 to 30) in the en bloc resection group. CONCLUSIONS: En bloc resection with reconstruction had a lower rate of recurrence, but a higher rate of complication and poor functional results. Intralesional excision, combined with a local adjunctive application and packed with bone grafting or methylmethacrylate cement, was an acceptably good method with satisfactory results, which either decreased local recurrence or retained the function of the joint. PMID- 9216121 TI - Interferon-gamma in cerebrospinal fluid of children with aseptic meningitis. AB - BACKGROUND: Certain cytokines may contribute to the sequence of events that lead to meningeal inflammation in bacterial meningitis. However, their role in viral meningitis is not so less well defined. We determined the cytokines levels in the cerebrospinal fluid (CSF) of children with aseptic meningitis and discussed their relationship with clinical and laboratory findings. METHODS: We determined the concentrations of interleukin-1 beta (IL-1 beta), interferon-gamma (IFN-gamma) and granulocyte-macrophage colony-stimulating factor (GM-CSF) in the CSF of 62 patients with aseptic meningitis including 17 patients with culture-proved enteroviral meningitis, and from 19 control acute febrile patients without meningitis. RESULTS: The GM-CSF in the cerebrospinal fluid was detected from one of the 62 patients with aseptic meningitis and none of the 19 controls. Fourteen (23%) of the 62 patients with aseptic meningitis and 2 (10.5%) of 19 controls had detectable IL-1 beta. There was no significant difference in IL-1 beta levels between patients with aseptic meningitis (4.4 +/- 11.4 pg/ml) and control group (2.4 +/- 7.7 pg/ml). The CSF IFN-gamma level was detectable in 40 (65%) of 62 patients and 6 (31.6%) of 19 controls. The mean CSF IFN-gamma concentration was significantly higher in patients with aseptic meningitis when compared with that in control group (37.9 +/- 48.8 pg/ml vs 17.5 +/- 29.7 pg/ml; p = 0.007). CONCLUSIONS: IFN-gamma was detectable in the CSF in 65% of patients with aseptic meningitis and the role of interferon-gamma remains to be determined. PMID- 9216122 TI - Wide opening method for vocal fold retention cyst. AB - BACKGROUND: Vocal fold cyst is a common cause of dysphonia. In our reported series, most cases were retention cysts. Enucleation of the cyst under microlaryngoscopy is usually considered to be an ideal treatment despite the fact that cyst rupture is frequently encountered with this procedure. In this report, we present a wide opening method for vocal fold retention cyst. METHODS: Twenty consecutive patients with vocal fold retention cysts larger than 2 mm in diameter were operated upon using the wide opening method (marsupialization). The medial cyst wall was excised along with the overlying mucosa, while the lateral cyst wall was preserved on the vocal fold. The cyst was widely opened following this procedure. RESULTS: Perceptual voice improvement was noted postoperatively. Videostroboscopy and acoustic analysis were also applied to confirm the perceptual results. CONCLUSIONS: The wide opening method has the advantages of simplicity, minimal tissue injury, rapid functional recovery and low recurrence. This technique can be considered another standard treatment of choice for medium- or large-sized vocal fold retention cysts. PMID- 9216123 TI - Presumptive identification of streptococci by pyrrolidonyl-beta-naphthylamide (PYR) test. AB - BACKGROUND: Group A streptococci and enterococci can be differentiated from other streptococci on the basis of their ability to cleave L-pyrrolidonyl-beta naphthylamide. METHODS: In the present study, the L-pyrrolidonyl-beta naphthylamide (PYR) test, pigment medium and bile esculin medium have been used to presumptively identify the streptococci. In total, 114 strains of group A streptococci, 350 strains of non-group A streptococci, 202 strains of enterococci and 197 strains of non-enterococci have been tested. RESULTS: The results of the present investigation show that sensitivities of different test methods are: PYR broth, 99.08%; Murex PYR, 98.48%; bacitracin, 95%; bacitracin and sulfamethoxazole/trimethoprim (SXT), 95%; pigment medium, 99.23%; bile esculin medium, 99.26%. Additionally, specificities of various tests are: PYR broth and Murex PYR, 99.82%; bacitracin, 90.90%; bacitracin and SXT, 98.87%; pigment medium and bile esculin medium, 100%, respectively. CONCLUSIONS: PYR test has been observed to be very easy to use and may hence be considered as a rapid, reliable and cost-effective method for presumptive identification of group A streptococci and enterococci in the clinical laboratory. PMID- 9216124 TI - Malignant epithelial neoplasm consistent with primitive cystic hepatic neoplasm with mesothelial differentiation: a case report. AB - Mesothelioma are primary tumors of the celiomic cavity and are seen more often in adults than in children: only an estimated 2-5% of all cases present within the first two decades of life. To best knowledge of the reviewing world literature reported to date, no more than 80 proved cases of this tumor have occurred in children. One-third of mesothelioma originate in the peritoneum and two-thirds arise in the pleural cavity. Mesothelioma of the liver are extremely rare; a review of the English literature shows only three adult cases that have been reported as fibrous mesothelioma of the liver; experience with these cases suggests a high potential for recurrence, but no progression to malignancy. Cystic mesothelioma occur mainly in adults and are considered to be benign and curable. We describe a case of malignant epithelial neoplasm consistent with primitive cystic hepatic neoplasm with mesothelial differentiation arising in a 3 year-old boy, a condition which has never before been reported in childhood. Malignant primitive cystic mesothelioma is possible that some cases of intraabdominal mesenchymoma or hamartoma with malignant differentiation may have been misdiagnosed in the past; future cases should be fully evaluated, to establish the true incidence of mesothelioma disease in children. PMID- 9216125 TI - Cutaneous Rosai-Dorfman disease manifestating as recurrent breast tumor: a case report. AB - A case of cutaneous Rosai-Dorfman disease (sinus histiocytosis with massive lymphadenopathy) manifestating as a recurrent breast tumor is reported. The tumor occurred on the left breast of a 35-year-old woman. Before arriving at the correct diagnosis, four biopsies had been performed with various diagnoses of chronic inflammation, plasma cell mastitis and inflammatory pseudotumor. Numerous typical histiocytes with lymphophagocytosis appeared in the final excised specimen, and a correct diagnosis was made. Ultrastructural examination revealed no evidence of Birbeck granule. The literature concerning Rosai-Dorfman disease manifestating as breast tumor is reviewed. Since the diagnosis is often overlooked in the absence of lymphadenopathy, a high index of suspicion is required to recognize this rare cutaneous Rosai-Dorfman disease. PMID- 9216126 TI - The design of a ferrofluid magnetic pipette. AB - An electromagnetic pipette using a ferrofluid was designed to sample liquid volumes smaller than 0.2 microliter. Submicroliter sample sizes are desirable for reducing the amount of costly reagents and reducing sample requirement for large scale analysis. The pipette consists of four electromagnets arranged such that air-gaps are aligned to accommodate a tube. A light-hydrocarbon-based ferrofluid is contained in the tube and acts as a plunger. The position of the ferrofluid in the tube was controlled to within 0.2 mm by combining adjacent air-gap magnetic fields. The position of the ferrofluid as a function of time and magnetic pressure as a function of position was measured in one electromagnet air-gap from the device. Maximum pressure measured was 770 Pa which corresponds to a maximum velocity of 0.9 cm/s. The assembled pipette weighs approximately 25 g, and it measures 4 cm long, 1 cm wide, and 3 cm high. PMID- 9216127 TI - A triaxial accelerometer and portable data processing unit for the assessment of daily physical activity. AB - The present study describes the development of a triaxial accelerometer (TA) and a portable data processing unit for the assessment of daily physical activity. The TA is composed of three orthogonally mounted uniaxial piezoresistive accelerometers and can be used to register accelerations covering the amplitude and frequency ranges of human body acceleration. Interinstrument and test-retest experiments showed that the offset and the sensitivity of the TA were equal for each measurement direction and remained constant on two measurement days. Transverse sensitivity was significantly different for each measurement direction, but did not influence accelerometer output (< 3% of the sensitivity along the main axis). The data unit enables the on-line processing of accelerometer output to a reliable estimator of physical activity over eight-day periods. Preliminary evaluation of the system in 13 male subjects during standardized activities in the laboratory demonstrated a significant relationship between accelerometer output and energy expenditure due to physical activity, the standard reference for physical activity (r = 0.89). Shortcomings of the system are its low sensitivity to sedentary activities and the inability to register static exercise. The validity of the system for the assessment of normal daily physical activity and specific activities outside the laboratory should be studied in free-living subjects. PMID- 9216129 TI - A real-time microprocessor QRS detector system with a 1-ms timing accuracy for the measurement of ambulatory HRV. AB - The design, test methods and results of an ambulatory QRS detector are presented. The device is intended for the accurate measurement of heart rate variability (HRV) and reliable QRS detection in both ambulatory and clinical use. The aim of the design work was to achieve high QRS detection performance in terms of timing accuracy and reliability, without compromising the size and power consumption of the device. The complete monitor system consists of a host computer and the detector unit. The detector device is constructed of a commonly available digital signal processing (DSP) microprocessor and other components. The QRS detection algorithm uses optimized prefiltering in conjunction with a matched filter and dual edge threshold detection. The purpose of the prefiltering is to attenuate various noise components in order to achieve improved detection reliability. The matched filter further improves signal-to-noise ratio (SNR) and symmetries the QRS complex for the threshold detection, which is essential in order to achieve the desired performance. The decision for detection is made in real-time and no search-back method is employed. The host computer is used to configure the detector unit, which includes the setting of the matched filter impulse response, and in the retrieval and postprocessing of the measurement results. The QRS detection timing accuracy and detection reliability of the detector system was tested with an artificially generated electrocardiogram (ECG) signal corrupted with various noise types and a timing standard deviation of less than 1 ms was achieved with most noise types and levels similar to those encountered in real measurements. A QRS detection error rate (ER) of 0.1 and 2.2% was achieved with records 103 and 105 from the MIT-BIH Arrhythmia database, respectively. PMID- 9216128 TI - Wavelength selection for low-saturation pulse oximetry. AB - Conventional pulse oximeters are accurate at high oxygen saturation under a variety of physiological conditions but show worsening accuracy at lower saturation (below 70%). Numerical modeling suggests that sensors fabricated with 735 and 890 nm emitters should read more accurately at low saturation under a variety of conditions than sensors made with conventionally used 660 and 900 nm band emitters. Recent animal testing confirms this expectation. It is postulated that the most repeatable and stable accuracy of the pulse oximeter occurs when the fractional change in photon path lengths due to perturbations in the tissue (relative to the conditions present during system calibration) is equivalent at the two wavelengths. Additionally, the penetration depth (and/or breadth) of the probing light needs to be well matched at the two wavelengths in order to minimize the effects of tissue heterogeneity. At high saturation these conditions are optimally met with 660 and 900 nm band emitters, while at low saturation 735 and 890 nm provide better performance. PMID- 9216130 TI - Linear and nonlinear ARMA model parameter estimation using an artificial neural network. AB - This paper addresses parametric system identification of linear and nonlinear dynamic systems by analysis of the input and output signals. Specifically, we investigate the relationship between estimation of the system using a feedforward neural network model and estimation of the system by use of linear and nonlinear autoregressive moving-average (ARMA) models. By utilizing a neural network model incorporating a polynomial activation function, we show the equivalence of the artificial neural network to the linear and nonlinear ARMA models. We compare the parameterization of the estimated system using the neural network and ARMA approaches by utilizing data generated by means of computer simulations. Specifically, we show that the parameters of a simulated ARMA system can be obtained from the neural network analysis of the simulated data or by conventional least squares ARMA analysis. The feasibility of applying neural networks with polynomial activation functions to the analysis of experimental data is explored by application to measurements of heart rate (HR) and instantaneous lung volume (ILV) fluctuations. PMID- 9216131 TI - A dynamic neuromuscular model for describing the pendulum test of spasticity. AB - Both dynamic and static thresholds, as well as the gain in the stretch reflex loop, affect the sensitivity of motoneurons to muscle stretch. How the variation in each parameter will influence the mechanical behavior of patients with spasticity is not well understood because of the difficulty in experimentally isolating individual parameters. A neuromuscular dynamic model, based on the pendulum test of spasticity, has been developed to study the specific contribution of individual parameter abnormalities in stretch reflex loops to the observed mechanical abnormalities. The model contains detailed nonlinear dynamics of muscle force generation and stretch reflexes. A computer simulation of the model indicates that the stretch reflex thresholds and the gain have different influences on the leg swing in the pendulum test of spasticity. Individual changes in the static stretch reflex threshold, in the dynamic threshold, or in the gain can not stimulate the whole spectrum of spasticity severity. When simultaneous changes in all three parameters of the stretch reflex loop occur, a small variation of the gain coupled with changes in both static and dynamic thresholds can produce increasing severity of spasticity as the thresholds further decrease. The model is also successful in simulating the effect of posture changes on spasticity. PMID- 9216134 TI - Determination of the red blood cell ability to traverse cylindrical pores. AB - An expression relating the red blood cell (RBC) volume and membrane surface area to the pore minimum radius /maximum length which the cell is able to traverse is derived. The application of this analytical model to design/specification of filters for RBC deformability evaluation is presented. The passage of the RBC's through relatively long (10 microns or more) submicron radii pores, which the developed model demonstrates, is also discussed. PMID- 9216132 TI - Closed-loop drug infusion for control of heart-rate trajectory in pharmacological stress tests. AB - The diagnosis of coronary artery disease (CAD) is an important task in the management of cardiology patients. Recently, the use of pharmacological stress testing has become available as an alternative to exercise stress testing (ETT). A new system (device-drug combination) was developed specifically for the diagnosis of coronary artery disease. The system uses a novel catecholamine, arbutamine, which is infused intravenously to increase heart rate (HR) and cardiac contractility in order to evoke signs of ischemia. The development of a closed-loop control algorithm for the delivery of this drug and a pharmacodynamic (PD) model representing the HR response to arbutamine infusions are presented. Model parameters are estimated from clinical data on normal volunteers and patients. Based on this mathematical model, a rule-based control algorithm is designed. The structure of the control algorithm is discussed and testing of the algorithm based on simulations and animal and human trials are summarized. Results from clinical trials shows that the algorithm controls the HR increase according to a selected trajectory. The automated delivery of the drug can provide the cardiologist with an efficient, effective, and safe method for administering a pharmacological stress test. PMID- 9216133 TI - Sensitivity distributions of EEG and MEG measurements. AB - It is generally believed that because the skull has low conductivity to electric current but is transparent to magnetic fields, the measurement sensitivity of the magnetoencephalography (MEG) in the brain region should be more concentrated than that of the electroencephalography (EEG). It is also believed that the information recorded by these techniques is very different. If this were indeed the case, it might be possible to justify the cost of MEG instrumentation which is at least 25 times higher than that of EEG instrumentation. The localization of measurement sensitivity using these techniques was evaluated quantitatively in an inhomogeneous spherical head model using a new concept called half-sensitivity volume (HSV). It is shown that the planar gradiometer has a far smaller HSV than the axial gradiometer. However, using the EEG it is possible to achieve even smaller HSV's than with whole-head planar gradiometer MEG devices. The micro superconducting quantum interference device (SQUID) MEG device does have HSV's comparable to those of the EEG. The sensitivity distribution of planar gradiometers, however, closely resembles that of dipolar EEG leads and, therefore, the MEG and EEG record the electric activity of the brain in a very similar way. PMID- 9216135 TI - Efficient computation of amplitude and phase maps in nuclear medicine equilibrium gated cardiac studies. AB - The Goertzel algorithm is proposed as a method to obtain the first harmonic coefficient of time activity curves from equilibrium gated cardiac studies. The coefficients are used to produce functional images. The algorithm achieves an important reduction in the number of operations and memory accesses needed to compute the coefficients. PMID- 9216136 TI - Estimation of slowly changing components of physiological signals. AB - A method for the estimation of slowly changing components of physiological signals is presented in this communication. The method is based on a sequential approximation of slowly changing components by a low-order polynomial function. The polynomial coefficients are obtained by minimizing the distance between the expected zero crossing density (ZCD) value of the fast components of the physiological signal and the estimated ZCD value of these components. The method has been tested and preliminary results were satisfactory. PMID- 9216137 TI - The electrical conductivity of human cerebrospinal fluid at body temperature. AB - The electrical conductivity of human cerebrospinal fluid (CSF) from seven patients was measured at both room temperature (25 degrees C) and body temperature (37 degrees C). Across the frequency range of 10 Hz-10 kHz, room temperature conductivity was 1.45 S/m, but body temperature conductivity was 1.79 S/m, approximately 23% higher. Modelers of electrical sources in the human brain have underestimated human CSF conductivity by as much as 44% for nearly two decades, and this should be corrected to increase the accuracy of source localization models. PMID- 9216138 TI - A volume-conduction analysis of magnetic stimulation of peripheral nerves. AB - Magnetic stimulation is a method to study several nervous disorders as well as the intact nervous system in humans. Interest in magnetic stimulation of peripheral nerves has grown rapidly, but difficulties in locating the site of excitation have prevented it from becoming a routine clinical tool. It has been reasoned that the activating function of long and straight nerves is the first spatial derivative of the electric field component parallel to the nerves. Therefore, to predict the site of activation, one has to compute this field feature. We describe here an analytical mathematical model and investigate the influence of volume-conductor shape on the induced field. Predictions of the site of activation are given for typical stimulation coil arrangements and these results are compared with experimental and literature data. Comparisons suggest that the activating function is not simply the spatial gradient of the induced electric field, but that other mechanisms are also involved. The model can be easily utilized in the search for more efficient coil constructions and improved placements with respect to the target nerves. PMID- 9216139 TI - Numerical solution of the potential due to dipole sources in volume conductors with arbitrary geometry and conductivity. AB - The integral conservation equation for biological volume conductors with general geometry and arbitrary distribution of electrical conductivity is solved using a finite volume method. An effective conductivity was defined for the boundaries between regions with abrupt change of the conductivity to allow the simultaneous solution of the entire domain although the derivatives are not continuous. The geometrical singularities arising from the spherical topology of the coordinate system are removed using the conservation law. The resulting finite volume solution method is efficient both in central processing unit (CPU) time and memory requirements, allowing the solution of the volume conductor equation using a large number of mesh points (of the order of 10(5)) even on small workstations (like SGI Indigo). It results in very accurate solutions, as several comparisons with analytical solutions of head models reveal. The proposed finite volume method is an attractive alternative to the finite element and boundary element methods that are more common in bioelectric applications. PMID- 9216141 TI - Design criterions for active shielding of inhomogeneous magnetic fields for biomagnetic applications. AB - General analytical expressions for the field attenuation and the reaction factor for a spherical active compensated cabin are theoretically derived. The shielding effect of various materials and their thickness on external magnetic disturbances as well as the retroactive effect on the locally generated compensating fields of the compensating coils are then analyzed. A numerical evaluation of the analytical expressions developed is made and directives for practical measures are derived. Comparison with the experimental results obtained from the measurement of shielding properties of an equivalent cubic cabin is made. The so determined design criterions are then discussed in detail. PMID- 9216140 TI - Effect of source location on the scalp potential asymmetry in a numerical model of the head. AB - The correlation between source asymmetry in the brain and the potential amplitude asymmetry on the scalp was studied by a two-dimensional (2-D) numerical model of the head. The model employed computerized tomography (CT) images to define the different compartments of the head. The source was modeled by a dipole layer in the occiput for an occipital source (visual evoked potential generators) or a dipole layer around the cortex representing spontaneous activity generators. The volume conductor equation for the potential distribution was solved numerically using a finite volume method for two CT images; one had relatively symmetric left right anatomy while the other had a falx deviation of 6 degrees between the occiput and the nasion-inion line. By examining several arrangements of sources, it has been demonstrated that source asymmetry can cause nonnegligible asymmetry in the potential amplitude at the homotopic points on the scalp. This asymmetry, that is not related to real physiologic or psychological origin, should be taken into consideration in any EEG potential distribution analysis. PMID- 9216142 TI - Geometric approach for coupling enhancement of magnetically coupled coils. AB - This paper presents a geometric approach for the enhancement of the coupling coefficient between two magnetically coupled coils. It is demonstrated that the coupling coefficient can be considerably enhanced, if the turns of the coils are not concentrated at the circumferences, but distributed across the diameters. For analysis, each of the two coils is assumed to be composed of concentric circular loops. The experimental results are in very good agreement with the theoretical results. PMID- 9216143 TI - Input impedance and reflection coefficient in fractal-like models of asymmetrically branching compliant tubes. AB - A mathematical model is described, based on linear transmission line theory, for the computation of hydraulic input impedance spectra in complex, dichotomously branching networks similar to mammalian arterial systems. Conceptually, the networks are constructed from a discretized set of self-similar compliant tubes whose dimensions are described by an integer power law. The model allows specification of the branching geometry, i.e., the daughter-parent branch area ratio and the daughter-daughter area asymmetry ratio, as functions of vessel size. Characteristic impedances of individual vessels are described by linear theory for a fully constrained thick-walled elastic tube. Besides termination impedances and fluid density and viscosity, other model parameters included relative vessel length and phase velocity, each as a function of vessel size (elastic nonuniformity). The primary goal of the study was to examine systematically the effect of fractal branching asymmetry, both degree and location within the network, on the complex input impedance spectrum and reflection coefficient. With progressive branching asymmetry, fractal model spectra exhibit some of the features inherent in natural arterial systems such as the loss of prominent, regularly-occurring maxima and minima; the effect is most apparent at higher frequencies. Marked reduction of the reflection coefficient occurs, due to disparities in wave path length, when branching is asymmetric. Because of path length differences, branching asymmetry near the system input has a far greater effect on minimizing spectrum oscillations and reflections than downstream asymmetry. Fractal-like constructs suggest a means by which arterial trees of realistic complexity might be described, both structurally and functionally. PMID- 9216144 TI - Reduction of computational complexity in the butterfly search technique. AB - In the butterfly search technique, echoes from repeated firings of a transducer are resampled along a set of predetermined trajectories of constant velocities, called "butterfly lines," because of their intersection and crossing at a reference range. The slope of the trajectory on which the sampled signals satisfy a predetermined criterion appropriate for the type of signal in question, provides an estimate of the velocity of the target. The search for this trajectory is called "butterfly search," which can be carried out efficiently in a parallel processing scheme. The estimator can be based on the radio frequency (RF) A-lines, the envelopes, or the quadrature components. The butterfly search on quadrature components has shown outstanding noise immunity, even with relatively few successive scan lines, and was found to outperform all the common time domain and Doppler techniques in simulations and experiments with strong noise. It can be simply implemented using elementary digital signal processing hardware. However, it is possible to further improve upon its computational complexity to make the technique even simpler to implement, without any complex multipliers in the parallel channels. In this paper, we present some modifications that significantly reduce the computational complexity of butterfly search on quadrature components. PMID- 9216145 TI - Spectral composition of heart sounds before and after mechanical heart valve implantation using a modified forward-backward Prony's method. AB - This paper investigates the impact of the lung-thorax and heart-valve system on the overall spectral composition of the externally recorded heart sounds. The study concentrates in the case of the first and the second heart sounds for normal patients and patients before and after implantation of a mechanical valve in the mitral or aortic position. The analysis is performed using a modified forward-backward overdetermined Prony's method (MFBPM) which uses a forward backward mean filter and a modified procedure for estimating the position of the signal poles. In terms of the normalized cross-correlation coefficient, this method has an average modeling accuracy of 99.62% for representing the first and second heart sounds and an average least square time-domain error of 0.43%. Results obtained from 40 subjects show that the condition of the native mitral or aortic valve affects mostly the distribution of the amplitudes of the spectral components, whereas the number of the spectral components or their respective relative energy remains more or less unchanged. It has been found that the amplitudes of frequency components in the range 120-250 Hz are more affected by abnormalities of native mitral valves. Furthermore, in the case of the second heart sound the region 250-400 Hz has been found to be more affected by abnormalities in the aortic valve. It has also been found that the mechanical prosthetic heart valve affects mostly the spectrum beyond 400 Hz. A clear difference has been observed in the frequency spectrum above 400 Hz between both normally and abnormally functioning native valves and normally functioning mechanical valves. Preliminary results in some malfunctioning cases of mechanical prosthesis suggest that spectral components beyond 400 Hz can be used to monitor the condition of these prostheses. PMID- 9216146 TI - Detection of seizures from small samples using nonlinear dynamic system theory. AB - The electroencephalogram (EEG), like many other biological phenomena, is quite likely governed by nonlinear dynamics. Certain characteristics of the underlying dynamics have recently been quantified by computing the correlation dimensions (D2) of EEG time series data. In this paper, D2 of the unbiased autocovariance function of the scalp EEG data was used to detect electrographic seizure activity. Digital EEG data were acquired at a sampling rate of 200 Hz per channel and organized in continuous frames (duration 2.56 s, 512 data points). To increase the reliability of D2 computations with short duration data, raw EEG data were initially simplified using unbiased autocovariance analysis to highlight the periodic activity that is present during seizures. The D2 computation was then performed from the unbiased autocovariance function of each channel using the Grassberger-Procaccia method with Theiler's box-assisted correlation algorithm. Even with short duration data, this preprocessing proved to be computationally robust and displayed no significant sensitivity to implementation details such as the choices of embedding dimension and box size. The system successfully identified various types of seizures in clinical studies. PMID- 9216147 TI - Effect of high-power microwave on indicator bacteria for sterilization. AB - According to the superiority sterilization, a specially-designed microwave disinfector using high-power microwave energy was made. A series of sterilizing experiments have been made to determine the effect of microwave energy on several typical indicator bacteria such as Bacillus subtilis var. niger, Bacillus stearothermophilus, Bacillus pumilus E601, Staphylococcus aureus, Bacillus cereus. Under the conditions of different sterilization duration and unequal intensity of microwave power irradiation onto the bacteria, a useful result of killing bacteria has been observed, i.e., the Bacillus subtilis can be considered as an optimum indicator bacterium for microwave sterilization. PMID- 9216148 TI - Effects of record length selection on the accuracy of spectral estimates of heart rate variability: a simulation study. AB - To evaluate the effects of record length selection on the accuracy of spectral estimates of heart rate variability (HRV), a simulation study was carried out using a set of 58 signals obtained by autoregressive (AR) fitting a representative sample of real HRV signals. Four record lengths of 180, 300, 420, and 540 s were considered. Spectral estimation was performed by both the Blackman Tukey (B-T) and AR methods. Accuracy was assessed for: 1) point spectral estimates, by computing the normalized averaged bias (NAB) and variance (NAV); and 2) the most commonly used spectral parameters [total power (TP) and the powers in the bands: very low frequency (VLF) (0 divided by 0.04 Hz), low frequency (LF) (0.04 divided by 0.15 Hz), and high frequency (HF) (0.15 divided by 0.45 Hz)], by computing the normalized bias (NB) and variance (NV). The results are: whatever the record length considered, the 90th percentiles (90P) of the NAB were < 10%, whereas those of the NB were < 9% for TP, LF, and HF powers, and < 14% for the VLF power, in both methods. The NAV was proportional to the reciprocal of record length, showing high 90P values for the shortest record length (26.4% for B-T and 44.2% for AR). The NV showed the same trend but 90P values were much lower (< 8% for TP, LF, and HF powers and < 19% for VLF power, in both methods). In the final part of the paper a procedure for the computation of approximate upper bounds of the relative absolute error of spectral measures at each record length, based on the knowledge of the NB and NV, is presented. PMID- 9216149 TI - A mathematical study of some biomechanical factors affecting the oscillometric blood pressure measurement. AB - A mathematical lumped parameter model of the oscillometric technique for indirect blood pressure measurement is presented. The model includes cuff compliance, pressure transmission from the cuff to the brachial artery through the soft tissue of the arm, and the biomechanics of the brachial artery both at positive and negative transmural pressure values. The main aspects of oscillometry are simulated i.e., the increase in cuff pressure pulsatility during cuff deflation maneuvers, the existence of a point of maximum pulsations (about 1.5 mmHg) at a cuff pressure close to mean arterial pressure, and the characteristic ratios for cuff pressure pulsatility at systole and diastole (0.52 and 0.70, respectively, with this model, using basal parameters and an individual set of data for the arterial pressure waveform). Subsequently, the model is used to examine how alterations in some biomechanical factors may prejudice the accuracy of pressure measurement. Numerical simulations indicate that alterations in wall viscoelastic properties and in arterial pressure pulse amplitude may significantly affect the accuracy of pressure estimates, leading to errors as great as 15-20% in the computation of diastolic and systolic arterial pressure. By contrast, changes in arterial pressure mean value and cuff compliance do not seem to have significant influence on the measurement. Evaluation of mean arterial pressure through a characteristic ratio is not robust and may lead to misleading results. Mean arterial pressure may be better evaluated as the lowest pressure at which cuff pulse amplitude reaches a plateau. The obtained results may help to explain the nature of errors which usually limit the reliability of arterial pressure measurement (for instance in the elderly). PMID- 9216150 TI - Analysis of cerebral blood flow autoregulation in neonates. AB - The dynamic response of cerebral autoregulation to spontaneous changes in arterial blood pressure (ABP) is described by the relationship between cerebral blood flow velocity (CBFV) and resistance-area product (RAP). CBFV was measured with Doppler ultrasound in the middle cerebral artery and ABP with an intra arterial catheter in 66 neonates. Spontaneous changes in mean ABP were automatically detected and the maximum derivative was used to synchronize the coherent averaging of corresponding CBFV and RAP transients. These were classified into two groups corresponding to intact (group A) or impaired (group B) autoregulation. The cross correlation between RAP and CBFV indicates a significant relationship with a time delay of 5 s for group A. The frequency response of RAP was estimated by the cross spectra with CBFV. Groups A and B present a similar amplitude spectra but the phase spectra of group A lags that of group B. The impulse responses of the two groups are also markedly different and were used to simulate the velocity response to a 5% step change in ABP. Impulse responses were also obtained for four different levels of pCO2 showing that hypercapnia leads to an impulse response similar to that of group B (impaired autoregulation). This method can be used to extend the usual dichotomic classification adopted in clinical studies of autoregulation. PMID- 9216151 TI - Selective discrete Fourier transform algorithm for time-frequency analysis: method and application on simulated and cardiovascular signals. AB - The Selective Discrete Fourier transform (DFT) Algorithm [SDA] method for the calculation and display of time-frequency distribution has been developed and validated. For each time and frequency, the algorithm selects the shortest required trace length and calculates the corresponding spectral component by means of DFT. This approach can be extended to any cardiovascular related signal and provides time-dependent power spectra which are intuitively easy to consider, due to their close relation to the classical spectral analysis approach. The optimal parameters of the SDA for cardiovascular-like signals were chosen. The SDA perform standard spectral analysis on stationary simulated signals as well as reliably detect abrupt changes in the frequency content of nonstationary signals. The SDA applied during a stimulated respiration experiment, accurately detected the changes in the frequency location and amplitude of the respiratory peak in the heart rate (HR) spectrum. It also detected and quantified the expected increase in vagal tone during vagal stimuli. Furthermore, the HR time-dependent power spectrum displayed the increase in sympathetic activity and the vagal withdrawal on standing. Such transient changes in HR control would have been smeared out by standard heart rate variability (HRV), which requires consideration of long trace lengths. The SDA provides a reliable tool for the evaluation and quantification of the control exerted by the Central Nervous System, during clinical and experimental procedures resulting in nonstationary signals. PMID- 9216152 TI - Neural network analysis of flow cytometry immunophenotype data. AB - Acute leukemia is one of the leading malignancies in the United States with a mortality rate strongly influenced by the phenotype. This phenotype is based on detection of cell associated antigens normally expressed during leucopoietic differentiation. In this regard, leukemia classified as lymphoid or myeloid by phenotype is also classified as a candidate for the corresponding chemotherapy protocol. Additionally, the subtype of leukemia based on the degree of differentiation and cell maturity influence prognosis, response to treatment, and median survival times. In this paper, we analyze immunophenotype flow cytometry data toward categorization of leukemia into subcategories based on lineage and differentiation antigen expression. Twenty-eight inputs (derived from the mean fluorescence intensity of up to 27 antibodies, and an additional binary input denoting the past diagnosis of leukemia) are used as input to a neural classifier to categorize a total of 170 cases into the lineage and differentiation categories of leukemia. The neural classifier consisted of a feed forward network trained using back propagation. A complexity regulation term (weight decay) was used to improve the generalization performance of the neural classifier. A training error of 0.0% and a generalization error of 10.3% was obtained for categorization based on lineage, while a training error of 0.0% and a generalization error of 10.0% was obtained for categorization based on differentiation. These results indicate that objective classification of multifaceted phenotypes in leukemia can be achieved for analyzing multiparameter data in flow cytometry and further categorization into the prognostic subtypes. PMID- 9216153 TI - The time-sequenced adaptive filter for analysis of cardiac arrhythmias in intraventricular electrograms. AB - Implantable antitachycardia devices rely upon schemes for detecting cardiac arrhythmias which utilize rate and its variations; yet rate parameters often identify nonpathologic tachycardias as potentially dangerous and deliver unwarranted therapy. I have developed a predictive filter based upon the time sequenced adaptive algorithm to be used as a supplement to rate criteria for detecting and identifying serious arrhythmias. The method does not require a fixed template and is independent of a priori patient information. The algorithm also provides arrhythmia diagnosis immediately at the change in rhythm. Algorithmic parameters were determined based upon a training set of patient data, and performance of the technique was evaluated with a completely new test set of 20 arrhythmia passages. The new algorithm yielded a sensitivity and specificity for ventricular tachycardia of 91% and 82% and for ventricular fibrillation of 71% and 93%. Correlation waveform analysis was used to diagnose the same test set of arrhythmias. It yielded a sensitivity and specificity for ventricular tachycardia of 100% and 67% and for ventricular fibrillation of 50% and 100%. PMID- 9216154 TI - A novel family of compression algorithms for ECG and other semiperiodical, one dimensional, biomedical signals. AB - In this paper, a novel family of compression algorithms is presented, which is designed to exploit the redundancy of one-dimensional (1-D) semiperiodical biomedical signals resulting from the cyclic nature of the underlying physical process. The basic idea is that a pool of past-seen cycles is maintained and cycles to be encoded can be stored as transformed versions of those residing in the pool. Conceptually, this approach is an extension of dictionary-based coding schemes used for text compression to signal patterns residing in an n-dimensional space. A cycle transformation method is introduced in order to render the pattern matching process practical and to enable cycle substitution. Based on the principles of the algorithmic family and this transformation method, an electrocardiogram (ECG)-oriented algorithm is implemented and thoroughly tested. The performance of this implementation is examined theoretically and deductions about the optimal algorithm settings are made. The ECG compression algorithm is superior to the average beat subtraction algorithm as proposed by Hamilton and Tompkins in cases where high compression ratios are required. PMID- 9216155 TI - Determination of nerve conduction velocity distribution from sampled compound action potential signals. AB - The sampled compound action potential (CAP) data sequence was expressed as the circular convolution of the delay sequence and the sampled single fiber action potential (SFAP) data sequence. An algorithm, based on Hirose's method [1], was then developed to separate the delay sequence from the sampled CAP data sequence, and the nerve conduction velocity distribution (NCVD) was consequently calculated from the delay sequence. The NCVD was found to be the product of the amplitude of the SFAP and the number of fibers. Simulations show that the estimated results were in good agreement with the calculated results. Experiments were performed on ten sciatic nerves from five bullfrogs (Rana pipens) using two independent variables: interelectrode distance and stimulus current strength. The results estimated from CAP's recorded under each condition reflect the corresponding feature of NCVD of the condition. The advantage of the technique is to provide detailed information about both slow and fast conducting fibers. This technique also offers the possibility to directly calculate the nerve fiber diameter distribution from the sampled CAP data sequences. PMID- 9216156 TI - Dynamics of temperature dependent optical properties of tissue: dependence on thermally induced alteration. AB - Thermal damage in heated bovine myocardial tissue is assessed from measured changes in total reflection and transmission of light. Mathematical expressions, based on random walk analysis of light propagation within tissue slabs, are used to relate the diffuse reflection and transmittance to the absorption coefficient, mu a, and effective scattering coefficient, mu's, for samples of myocardial tissue which were subjected to rapid step changes in temperature. Time-dependent changes in mu's, indicate two processes, one with a fast and temperature dependent rate the other with a slow and apparently temperature-independent rate. For final temperatures above 56.8 degrees C and for the first 500 s after the temperature change, the optical parameters are well fit by exponential forms that exhibit temperature-dependent time constants as predicted by Arrhenius reaction rate theory of thermal damage. The scattering changes are associated with an apparent activation energy, delta E, of 162 kJ/mole and a frequency constant, A, of 3 x 10(23) s-1. This method provides a means for estimating optical coefficients which are needed to assess laser tissue dosimetry. PMID- 9216157 TI - A technique for identifying the total space or temperature dependent thermal parameters (TITP) of biological materials in vivo. AB - A new algorithm for simultaneously identifying the space or temperature dependent conductivity K, blood perfusion rate Wb, metabolic rate qm, and thermal diffusibility alpha in Pennes' equation is postulated. The obtained over determined equations are solved by way of Householder's transformation. The validity of the method is tested through an example. PMID- 9216158 TI - Position-selective activation of peripheral nerve fibers with a cuff electrode. AB - The degree of spatial selectivity which can be obtained with longitudinal dot tripoles in an insulating cuff was quantified in terms of the overlap between fiber populations activated by different tripoles. Previous studies have failed to take into account the relative influences of transverse current and longitudinal current on position-selective activation, and furthermore have not controlled for the differing sensitivities of large and small nerve fibers to electrical stimuli. In this study, these factors were taken into account. Transverse current from an anode positioned opposite the stimulating cathode was found to improve spatial selectivity, and selectivity was enhanced when the ratio of transverse current to longitudinal current was increased. Large fibers were excited before small fibers, irrespective of fiber position, indicating a combination of position and size selectivity. PMID- 9216159 TI - Quadrupolar stimulation for Cochlear prostheses: modeling and experimental data. AB - Cochlear implants are electrically driven in monopolar, bipolar, or common ground mode. Ideally, a quadrupolar mode is created with three colinear electrodes, where the outer poles are half the inverse polarity value of the center electrode. The resulting field is highly focused. Models of point sources show that the quadrupolar paradigm offers a greater choice of parameters to shape the field. Simulation with a lumped-parameter model of the cochlea confirms the focusing action of the quadrupole in the layers of the inner ear. Field measurements in saline solution and in the scala tympani of guinea pigs show that focusing occurs with the quadrupolar mode. It is conceivable that quadrupolar stimulation will affect the pitch place coding, reduce channel interaction and limit facial or tactile stimulation induced by current spread. PMID- 9216160 TI - Predictive pulmonary function of school children in an area of low air pollution in Taiwan. AB - The objective of this study was to determine the effect of different parameters on a predictive model of pulmonary function for elementary school children in an area of low air pollution in Taiwan. Healthy children aged 7 to 12 years from three elementary schools in low-air-pollution areas (Da-Chen, Mai-Liau and Tai Si) participated in the study. A total of 836 children (423 boys and 413 girls) were included in the study. During summer vacation, each child underwent a physical examination including a screening spirometry. A questionnaire regarding respiratory symptoms and indoor air pollutants was also completed by the children's parents. Air monitoring showed that the levels of outdoor pollutants were relatively low. Multiple linear regression analysis was performed with FVC (forced vital capacity) and FEV1 (forced expiratory volume in 1 sec) as dependent variables. Gender, age, height, weight, technician and indoor air pollution parameters were the independent variables. The results showed that gender,height, weight and technician were the most significant variables for predicting FVC and FEV1. The various indoor air pollution parameters seemed to have no influence on the pulmonary function of children, except that mildew in bedrooms mildly decreased FEV1. Regression analysis showed that all the pulmonary function parameters measured had a positive correlation with height, whereas weight correlated only with certain parameters. Because both indoor and outdoor air pollution was relatively low, we suggest that this model could be used as a basic predictive model of pulmonary function for elementary school children in Taiwan. PMID- 9216161 TI - Risk factors associated with coronary artery stenosis among patients with chest pain in eastern Taiwan. AB - To analyze the risk factors for coronary artery stenosis among patients who were admitted to the hospital with chest pain in eastern Taiwan, we retrospectively reviewed the clinical data of 444 patients who received coronary arteriography, including 62 aborigines. Results indicated that there were 268 patients (64%) with coronary stenosis. Male to female ratio, mean age, pack-years of cigarette smoking, and hypertension were significantly higher in patients with coronary stenosis (CS group) than in patients without coronary stenosis (control group). There were also significant differences in the serum levels of total cholesterol, low-density lipoprotein cholesterol and high-density lipoprotein cholesterol between the CS and control groups. The total cholesterol and low-density lipoprotein cholesterol of both the CS and control groups were lower than those reported in western Taiwan. The prevalence of coronary stenosis among Taiwanese patients who presented with chest pain was significantly higher than in aborigines (64% vs 38%), although there were no significant differences in common risk factors between these two ethnic groups. These results revealed that the common risk factors in eastern Taiwan were the same as those reported here and suggest that race may be an important risk factor for coronary stenosis. PMID- 9216163 TI - Penicillin-resistant pneumococcal infections in children. AB - Penicillin-resistant pneumococcal infections have been reported worldwide, but rarely reported in Taiwan. From 1990 to 1995, the rate of penicillin-resistant Streptococcus pneumoniae (PRSP) infections in our hospital increased from less than 10% during the first 2 years (1990-91) to 45% during the last 2 years (1994 95). From 1990 to 1995, twenty-four patients with systemic pneumococcal infections were diagnosed in the Department of Pediatrics at Mackay Memorial Hospital. Pneumococci were isolated from blood in 20 patients, cerebrospinal fluid in 12 patients, joint fluid in one patient and pleural effusion in one patient. Four patients had underlying diseases, including ileal atresia, Wiskott Aldrich syndrome, congenital heart disease, and perilymph fistula. Of the 24 isolates of S. pneumoniae, 17 (70.8%) were intermediately penicillin resistant (minimum inhibitory concentrations between 0.1 and 1.0 microgram/mL), and 7 (29.2%) were highly resistant (minimum inhibitory concentrations > 1.0 microgram/ml). Fourteen patients recovered completely, two had minor sequelae, two had major sequelae, and six died. Four of the 12 patients with meningitis died. In this study, both the rate of PRSP as well as the mortality of patients with PRSP meningitis were high, as compared to previous reports. To reduce the mortality and morbidity of systemic pneumococcal infections, the oxacillin disc diffusion test is important in addition to appropriate antibiotic therapy. PMID- 9216162 TI - Uptake of charged liposomes by the rat liver. AB - The influences of liposomal surface charge on hepatic uptake of liposomes in vivo and in vitro were studied in rats. 3H-inulin was entrapped in positively charged, negatively charged or neutral liposomes composed of phosphatidylcholine, cholesterol, and a charge lipid at a molar ratio of 4:4:1, or without a charge lipid. Plasma and liver concentrations of inulin were measured 5, 15, 30, and 60 minutes after an intravenous bolus dose of liposome. The in vitro study comprised incubation of liposomes with primary culture of isolated rat hepatocytes. The incubation was terminated at 30, 60, 90, 120, and 180 minutes, and cellular uptake of inulin was determined. The results showed that the positively charged liposomes resulted in the highest plasma liposome concentration. The negatively charged liposomes gave the highest liver liposome concentration, among the three liposomes. In vitro study also demonstrated that cellular uptake of a negatively charged liposome was higher than positively charged or neutral liposomes. These results suggest that positively charged liposomes might be preferable for maintaining higher plasma concentrations, while negatively charged liposomes might be more efficient for drug delivery to the liver. PMID- 9216164 TI - CATCH 22: deletion of locus 22q11 in velocardiofacial syndrome, DiGeorge anomaly, and nonsyndromic conotruncal defects. AB - DiGeorge anomaly (DGA) and velocardiofacial syndrome (VCFS) are frequently associated with monosomy of chromosomal subband 22q11. It is not clear whether individuals who present with only some of the features (e.g., isolated hypoparathyroidism or conotruncal defects) of these conditions also have the same deletion. In a prospective study of 30 children from 1994 to 1996, we used both high-resolution banding and fluorescence in situ hybridization (FISH) to assess the deletion status of children with a wide range of DGA-like or VCFS-like clinical features. Microdeletion of the chromosomal subband 22q11.22 was detected in 17 children by high-resolution banding and in two additional children with conotruncal defect (CTD) who had submicroscopic deletions proved by FISH analyses. Of the patients with microscopical deletion (n = 17), only six had classical DGA (n = 4) or VCFS (n = 2) phenotypes. The other 11 had various forms of congenital heart defects as the only presenting signs of deletion. One patient with DGA stigmata had another chromosomal aberration of monosomy 10p13. Only 10 patients were found to have neither cytogenetic nor molecular abnormalities. Therefore, it appeared that the majority, if not all, of the DGA and VCFS patients with the 22q11 deletion were identifiable using FISH with the single N25 (D22S75) probe. It would also be advisable that children with isolated CTD should be carefully examined to detect the other morphologic abnormalities of DGA and VCFS, or CATCH 22 (cardiac defects, abnormal facies, thymic hypoplasia/aplasia, cleft palate, hypocalcemia, and 22q11 deletion). Once these abnormalities are found, a molecular cytogenetic analysis for the so-called 22q11 region is indicated. Because of other associated chromosomal findings, routine or high resolution cytogenetic analysis should be performed on patients with suspected CATCH 22. PMID- 9216165 TI - Surgical treatment of congenital convex pes valgus. AB - Congenital convex pes valgus is a rare, complex and heterogeneous anomaly of the foot, which is difficult to treat. From 1985 to 1994, we treated 14 patients (20 feet) with this deformity. There were six boys and eight girls. Six patients had bilateral involvement. Their ages at the time of surgery ranged from 4 to 32 months (mean, 13 mo). In eight patients (10 feet), the etiology was arthrogryposis multiplex congenita. The eitiology was unknown in six patients (10 feet). Associated problems included hip dislocation in seven patients, flexion or extension contractures of the knee in six patients, and clubfoot deformity in three patients. One patient had undergone previous surgery for release of a knee contracture. To achieve a plantigrade and balanced foot, all patients had one stage surgical open reduction and circumferential release for the correction of deformities. At a minimum of 2 years following surgery, 11 patients (16 feet) had satisfactory results determined by a 10-point evaluation system based on both clinical and radiographic features. Satisfactory results and ambulatory prognosis were related to the etiology and severity of each patient's condition. Two patients (two feet) had scars with poor cosmetic appearance and two patients (three feet) had inadequate correction. We concluded that with adequate soft tissue release and complete talocalcaneonavicular joint reduction in one-stage surgery, proper postoperative maintenance, and physical therapy, satisfactory results in the treatment of congenital convex pes valgus can be achieved. PMID- 9216166 TI - Effect of electrical stimulation on callus maturation during callus distraction in rabbits. AB - Callus distraction is currently the most widely used technique for limb lengthening. Prolonged treatment time is its main shortcoming. In this study, we tested the effectiveness of electrical stimulation during various stages of lengthening in order to decrease the treatment time. Seventy-five New Zealand white rabbits, about 2 kg in body weight, were divided into five groups. All groups received single level tibial lengthening of 1 cm by callotasis using a mini-lengthener. Group 1 rabbits did not receive electrical stimulation and could move freely in their cages. Group 2 rabbits (sham control), with electrodes on their left legs, were restrained on a wooden frame for 8 hours every day without electrical stimulation. Group 3 rabbits received capacitively coupled electrical stimulation, 60 kHz, 500 mV, on the left leg for 8 hours every day during the distraction period. Group 4 rabbits received electrical stimulation during the neutralization period. Group 5 rabbits received electrical stimulation during both the distraction and neutralization period. All rabbits were restrained during electrical stimulation. Weekly radiographs were taken to determine the time of appearance of at least three neocortices in the lengthening callus. At that time, the fixators were removed. The healing indexes (total time in fixator divided by length gained, days/cm) in the five groups of rabbits were compared. The range of motion of the ipsilateral knees and ankles and complications of treatment were recorded. The results showed that electrical stimulation applied during leg lengthening by callus distraction can significantly reduce the treatment time and the healing indexes, but the electrical stimulation may contribute to decreased motion in the ipsilateral knee and ankle joints. PMID- 9216167 TI - Magnetic resonance imaging in patients with hemodialysis-related arthropathy. AB - From April 1993 to December 1993, we prospectively studied the knees of 15 patients (mean age, 48 +/- 11 yr) receiving long-term hemodialysis (mean duration, 9 +/- 4 yr) using magnetic resonance imaging (MRI) techniques including, T1 weighted spin-echo, multiplanar gradient recalled, and postcontrast T1 with chemical shift-selective, fat-saturation pulse sequences. Analysis of these images revealed that the three findings most indicative of hemodialysis related arthropathy were intramedullary, cortical and soft tissue lesions. Knee pain was significantly correlated with the presence of soft tissue lesions. Cortical lesions were usually associated with soft tissue lesions. Inflammatory changes adjacent to soft tissue lesions were demonstrated in postcontrast studies in all patients with soft tissue lesions. Increases in water content in those lesions appeared to increase the signal intensity. Our results indicate that MRI is useful in demonstrating the extent of hemodialysis-related arthropathy involvement, especially in hemodialysis patients suffering from knee pain. PMID- 9216168 TI - Automated perimetry in primary open-angle glaucoma. AB - Primary open-angle glaucoma (POAG) is one of the leading causes of blindness worldwide. Patients are usually asymptomatic until severe anatomic and functional damage occurs. Visual field examination is important in managing glaucoma patients, especially in the treatment and follow-up of moderate to advanced cases. This study was designed to investigate the patterns of automated visual field defects in POAG. We studied 296 POAG patients from the Glaucoma Clinic of the National Taiwan University Hospital from January 1986 to June 1996. The perimetry was performed using the Octopus program 32 or 38, or Humphrey program 30-2, and the results were analyzed according to the Glaucoma Hemifield Test design. The depth and frequency of locations of visual field defects were computed, and stratified by age and intraocular pressure (IOP). The results suggested that the most frequently affected locations in POAG were paracentral areas, while the deepest scotoma appeared in the upper and lower nasal areas. Patients with IOP higher than 25 mmHg or older than 50 years had higher mean sensitivity losses in both superior and inferior hemifields. The results of this study revealed the patterns of automated visual field defects in POAG may be a useful guide for the management of POAG patients. PMID- 9216169 TI - Predictive factors for early menarche in Taiwan. AB - The rapid increase of breast cancer in Taiwan has prompted the authors to evaluate the predictive factors of early menarche among contemporary Taiwanese girls. A total of 895 four-grade girls from eight elementary schools in Taipei City and County were identified as a closed cohort from the first semester of 1993. Data were collected from self-administered questionnaires and school records. A total of 799 girls who had not menstruated in the first year remained in the group through 1994. The effects of potential predictive factors were assessed by logistic regression. Among the 799 girls followed, 69 (8.6%) had first menstruation between the fourth and fifth grades. Height, weight, body mass index and maternal early onset of menarche were positively related to the onset of menarche within the preceding year. Energy consumption during exercise showed only moderate association after being adjusted for age and weight. Calorie intake from junk food was not associated with early menarche within the preceding year. Poor interpersonal family relationships and stressful life events also showed a moderate association with early menarche. The data obtained supported the hypothesis that height, weight, body mass index and maternal early menarche are positive predictive factors of early menarche. The effects of exercise and childhood stress are less prominent. PMID- 9216170 TI - Unusual manifestations in children with Kawasaki disease. AB - Between January 1983 and December 1992, the medical records of 187 patients (116 boys and 71 girls) with Kawasaki disease (KD) who were admitted to the hospital in the acute phase were retrospectively reviewed. Of these, 175 patients (93.6%) were under 4 years of age. Among the six principal symptoms of KD, the incidence of cervical lymphadenopathy (41.2%) was relatively low. Additionally, we found some unusual features including intussusception in a 4-month-old female, transient thrombocytopenia in seven children (3.7%) and isolated azotemia in five. KD is a systemic disease of unknown etiology. The diverse associated features make KD puzzling and difficult to diagnose. In caring for children with KD, physicians should be alert to the principal symptoms as well as the unusual associated manifestations. PMID- 9216171 TI - Tubo-ovarian abscess caused by multidrug resistant Bacteroides gracilis. AB - Bacteroides gracilis infections are very rare and have always been reported to have a polymicrobial etiology. The majority of these infections occur in the head and neck areas, the pleuropulmonary system, and the abdominal cavity. We report a case of tubo-ovarian abscess caused by B. gracilis. A literature search revealed no previous reports. Our patient, a 29-year-old woman, experienced fever and lower abdominal pain caused by a tubo-ovarian abscess. Her treatment consisted of surgical drainage and prolonged intravenous antibiotic therapy. Initial therapy with cefotaxime and metronidazole failed and she remained febrile after the laparotomy. Her clinical condition improved slowly after initiation of imipenem therapy. Culture of a pus specimen obtained during surgery yielded B. gracilis, which was resistant to imipenem but susceptible to clindamycin. Combination therapy with imipenem and clindamycin was then administered and she recovered completely. Clindamycin was subsequently prescribed for long-term bacterial suppression. The potential difficulties in treating B. gracilis infections were a major clinical concern in the treatment of this patient. PMID- 9216172 TI - Mixed meningococcal and tuberculous meningitis. AB - Meningococcal meningitis is one of the most common bacterial infections of the meninges worldwide, and tuberculous infection is the most common cause of chronic meningitis in Taiwan. However, mixed meningococcal and tuberculous meningitis is rare. We describe a 27-year-old woman with a case of culture-proven meningococcal and tuberculous meningitis verified by polymerase chain reaction on a cerebrospinal fluid specimen. The patient was initially treated with intravenous antibiotics including penicillin G and chloramphenicol. Though the patient responded well to therapy initially, her subsequent clinical deterioration was finally controlled by long-term antituberculous medications. PMID- 9216173 TI - Remotely controllable temporary afterloading brachytherapy for malignant brain tumors. AB - We present the results of six patients with recurrent malignant brain tumors who underwent an alternative system of interstitial brachytherapy. In each patient, the neurosurgeon initially inserted several small catheters into the tumor target through a stereotactic procedure or open craniotomy. Later, the patient was treated using an intracatheter temporary implant of a high-dose-rate 192Iridium. We delivered a mean dose of 6 Gy per fraction into the rim of the tumor margin and limited it to 2 Gy at a distance of 1 cm outwards. Each patient received one fraction, 3 to 5 minutes every 2 days, for a total of three fractions. In between these treatments, patients performed regular daily activities and had nursing care as usual. There was no surgical mortality in this study. One patient had late-onset anemia. This treatment modality has the advantage of combining cytodiagnosis/reduction and radiation within one very short therapeutic time period. PMID- 9216174 TI - Solitary ileal lipoma presenting with ileocolic intussusception: an unusual cause of enteritis cystica profunda. AB - We report a 63-year-old man who presented with a history of intermittent vague abdominal pain, diarrhea, and weight loss. Colonofibroscopic examination disclosed an ileocolic intussusception with a polypoid tumor on the leading edge of the intussusception. Both double contrast barium enema and computerized tomography of the abdomen confirmed this finding. The excised specimen had an ulcerated ileal lipoma with enteritis cystica profunda on the overlying epithelium. PMID- 9216176 TI - [Epidemiology of lung cancer in former poison-gas workers and molecular approaches to lung cancer]. AB - Former poison-gas workers were found to have a high incidence of respiratory neoplasia. The development of cancer in these people could be suppressed by the administration of Nocardia rubra cell-wall skeleton. Efforts to improve the prevention and treatment of lung cancer include studies of telomerase activity and genetic aberrations in samples from patients with cancer, monitoring of the concentration of anti-cancer drugs, and studies of the feasibility of gene therapy with adenovirus vectors. PMID- 9216175 TI - [Pathophysiology of acute lung injury]. AB - Almost all respiratory diseases except benign lung tumors and lung dysplasia entail acute lung injury. The many clinical conditions associated with acute lung injury include aspiration pneumonia, bacterial pneumonia, and sepsis. The fundamental cause of acute lung injury is pulmonary vascular hyperpermeability. Pulmonary vascular hyperpermeability can be attenuated by nitric oxide and cyclic GMP, and potentiated by oxygen radicals and elastase released from neutrophils. Adhesion molecule inhibition could become an effective therapy against acute lung injury, because the adhesion molecules are very important in the pathogenesis of this condition. Adhesion molecules could also be useful markers of disease activity in various lung diseases. Neutrophil elastase inhibitors may become important as therapeutic agents against acute exacerbations of idiopathic interstitial pneumonia, because this pathological condition is a type of acute lung injury. Similarly, N-acetyl cysteine could also become a useful therapeutic agent against idiopathic interstitial pneumonia, because it is a precursor of glutathione, which is the major antioxidant in the fluid lining of the bronchial epithelium. PMID- 9216177 TI - [Exercise-induced asthma]. AB - Exercise-induced asthma (EIA) is common in children. In our experience, the incidence of EIA in adults with asthma was 54.4% (37 of 68), and those with and without EIA similar in many ways. Anti-cholinergic drugs were effective in patients with central airway obstruction during episodes of EIA, but disodium cromoglycate protected 80% of EIA patients regardless of the site of airway obstruction. The relationship between chemical mediators and EIA remains controversial but our data show a close relationship between the production of neutrophil chemotactic factor and the severity of EIA. To investigate the mechanism of EIA, we used hyperpnea-induced bronchoconstriction in sensitized rabbits. In this model, bronchoconstriction followed inhalation of dry air regardless of temperature; there was a refractory period, and the bronchoconstriction was completely blocked by an anti-cholinergic drug. The results of studies of inhalation of hypertonic saline, hyperosomolar solutions and amiloride suggest that hyperpnea-induced bronchoconstriction is caused by degranulation of mast cells or by vagal stimulation secondary to changes in osmolarity and in sodium concentration in the airway surface, which result from water loss induced by inhalation of dry air. Vascular phenomena are probably not involved in EIA. PMID- 9216178 TI - [Pathophysiology of diffuse panbronchiolitis and progress of the therapy]. PMID- 9216179 TI - [Gene therapy--present status and future prospects]. PMID- 9216180 TI - The expression of adhesion molecule and ARDS. PMID- 9216182 TI - [Airway hyperresponsiveness and airway mucosal permeability]. AB - The relationship between airway mucosal permeability and airway hyperresponsiveness was examined with tachykinins, their selective antagonists, and superoxide dismutase in male Hartley guinea pigs. In animals with ozone induced airway inflammation, airway hyperresponsiveness and mucosal permeability increased concurrently but there was a time lag before the increase in airway vascular permeability. To study the role of tachykinins in the increases in mucosal permeability and in hyperresponsiveness, we used a neurokinin-receptor antagonist, CP-96345, and a neurokinin-2 receptor antagonist, SR-48968. CP-96345 had no significant effect, but SR-48968 reduced the increase in airway mucosal permeability; their effects on airway hyperresponsiveness were the opposite of their effects on airway permeability. Tachykinins themselves, both substance P and neurokinin A, significantly increased airway mucosal permeability. Superoxide dismutase, a scavenger enzyme of superoxide, reduced the ozone-induced airway hyperresponsiveness. These data suggest that the factors causing airway hyperresponsiveness differ from those that influence mucosal permeability, but it is possible that these pathophysiologic conditions are caused by the same substances or processes. PMID- 9216183 TI - [Meanings of the clinical parameters of airway hyperresponsiveness]. AB - Clinically, threshold concentrations of agonist causing 20% decrease in FEV1 (PC20 FEV1) or 35% decrease in airway conductance (PC35Gaw) are usually used as parameters of airway hyperresponsiveness. In this session, what do these parameters mean was discussed based on theoretical analysis, experimental data and clinical observations. Theoretically, using these parameters, it seems very difficult to distinguish whether hyperresponsiveness is due to the change in sensitivity or maximum responses. Experimental and clinical observations suggest that these parameters are affected by multiple factors, for example, genetic vs. acquired factors and functional vs. structural factors. There may be differences in the degree of contributions of each factors on clinical parameters of airway responsiveness between asthmatics and control, and mild and chronic severe asthma. Clinically, considering such differences may be important in interpreting the changes of these parameters. PMID- 9216184 TI - [Anti-inflammatory drugs for the treatment of bronchial hyperresponsiveness]. AB - To evaluate the effects of anti-inflammatory drugs on bronchial hyperresponse in patients with mild asthma, We examined inhaled beta 2-agonists, beclomethasone dipropionate (BDP) and suplatast tosilate. The results were as follows: 1. Inhaled beta 2-agonists, which have no direct anti-inflammatory effect, significantly improved bronchial hyperresponsiveness with inhalation of less than 6 puffs/day except tenoterol, but not with more than 6 puffs/day for two years. Patients with asthma therefore should use no more than 6 puffs/day of inhaled beta 2-agonists. Combined use of BDP and inhaled beta 2-agonists further improved bronchial hyperresponsiveness. This can be achieved by concurrent use of BDP, which has a potent anti-inflammatory effect. 2. Low dose BDP (200 micrograms/day), which is recommend in the Japanese guidelines for asthma treatment increased peak expiratory flow and improved bronchial hyperresponsivenss significantly during 8 weeks of treatment, but by 4 weeks after the end of BDP treatment, peak expiratory flow and bronchial hyperresponsiveness had worsened. Because we don't know the detail past history of patients with mild asthma, the patients must be treated with a relatively high dose of BDP (800 micrograms/day), which did improved bronchial hyperresponsiveness and significantly reduced the numbers of eosinophils and EG2 positive cells in the bronchial mucosa of patients with mild asthma. 3. Suplatast tosilate, which exerts its anti-inflammatory effect by suppressing IL-4 and IL-5 was given to patients with mild asthma for 6 weeks. Suplatast tosilate significantly increased peak expiratory flow and significantly improved bronchial hyperresponsiveness. It also signiticantly reduced the number of eosinophils and EG2 positive cells in the bronchial mucosa of eight patients with mild asthma. Therefore this drug may be useful for the treatment of patients with mild asthma. PMID- 9216185 TI - [Relation between the sialic acid/fucose ratio in airway secretions and the degree of airway remodeling in bronchial asthma--reversibility of airflow limitation by beta-agonists and airway remodeling in bronchial asthma]. AB - Airway remodeling caused by inflammation is believed to affect both airway hyperresponsiveness and the reversibility of airflow limitation. Airway remodeling entails changes in the bronchial secretion system. Mucin, one of the glycoprotein components of airway secretions, is a major product of submucosal glands and goblet cells. Airway remodeling causes both qualitative and quantitative changes in much production. We evaluated the relationship of airway remodeling, estimated by the change in glycoprotein in sputum, to airway hyperresponsiveness and to reversibility in patients with bronchial asthma. The %FEV1 after inhalation of a beta agonist was less in patients with severe asthma than in those with mild asthma. The sialic acid/fucose ratio in sputum correlated significantly with the slope of the dose-response curve during inhalation challenge, and with the %FEV1 after beta-agonist inhalation. These results suggest that the sialic acid/ fucose ratio in airway secretions reflects the degree of the airway remodeling in bronchial asthma. PMID- 9216186 TI - [Indices allowing early detection of chronic pulmonary emphysema]. AB - To establish criteria allowing early detection of pathologically significant alterations in pulmonary emphysema caused by smoking, pulmonary-function tests and high-resolution computed tomography were done in 104 subjects categorized into three groups: nonsmoking healthy adults, smokers with a normal FEV1%, and smokers with a low FEV1% (cross-sectional analysis). Fifty-six of the 104 patients underwent pulmonary-function testing and high-resdution computed fomography once per year for 3 years (longitudinal analysis). Cross-sectional and longitudinal analyses showed that abnormalities in functional residual capacity, in single-breath diffusing capacity for carbon monoxide, and in the average tomographic density of sections in the lower lung fields obtained after a deep inspiration could be used to predict whether the disease would reach an advanced stage, even if the patients had no significant symptoms at the time of testing. Relative areas of low-attenuation regions, which were alleged to directly reflect the size of emphysematous areas, appear not to be useful for early detection of pathological emphysema. PMID- 9216187 TI - [Clinical usefulness of helical computed tomography in evaluating chronic obstructive pulmonary diseases]. AB - To assess the usefulness of healical computed tomography (CT) in evaluating pathophysiological abnormalities in chronic obstructive pulmonary disease, we compared the results of helical CT with those of pulmonary-function tests in 46 subjects with pulmonary emphysema. We obtained 3-D images of emphysematous lung tissue by choosing voxels with values less than -930 HU because the mean CT score in 10 normal subjects was -869 +/- 29 HU. For each patient the total lung volume (TLV) was computed from the 3-D image of the entire lung with CT score less than 600 HU, and the volume of emphysematous lung tissue (ELV) was computed from the lung with CT score less than -930 HU. TLV measured on inspiration correlated significantly with TLC (r = 0.601, p < 0.001), and TLV measured on expiration correlated significantly with RV (r = 0.836, p < 0.0001). The difference between inspiratory TLV and expiratory TLV correlated significantly with VC (r = 0.781, p < 0.001). ELV both on inspiration and on expiration correlated significantly with FEV1/FVC (p < 0.001) and with RV (p < 0.01, p < 0.001, respectively). These results suggest that the volume data obtained by helical CT reflect functional abnormalities of the lung. In addition to volume data, the distribution of abnormal lung tissue is easily assessed by helical CT. We conclude that helical CT is useful in evaluating pathophysiological abnormalities in chronic obstructive pulmonary disease. PMID- 9216188 TI - [Neutrophil elastase and elastin-derived peptides in BAL fluid and emphysematous changes on CT scans]. AB - We examined the relationship between neutrophil elastase, elastin-derived peptides in bronchoalveolar lavage (BAL) fluid, and the development of pulmonary emphysema. The level of neutrophil elastase was higher in asymptomatic current smokers with emphysematous changes on computed tomographic scans than in current smokers without emphysematous changes, and was found to be correlated with the level of elastin-derived peptides in BAL fluid. Subjects with high levels of neutrophil elastase in BAL fluid had faster annual declines in FEV1. We conclude that the level of neutrophil elastase in BAL fluid can be used to differentiate asymptomatic cigarette smokers who are at risk for pulmonary emphysema from those who are not. PMID- 9216189 TI - [Pharmacologic therapy for patients with stable chronic obstructive pulmonary disease--emphasis on inhaled agents]. AB - One dose of an anticholinergic agents has about the same bronchodilatory effect as one of a beta 2-agonist. Anticholinergic agents may be used first in patients with stable chronic obstructive pulmonary disease, because of the possible adverse effects of beta 2-agonists. One puff of a metered-dose inhaler will not cause a maximal bronchodilatory effect. Both anticholinergic and beta-adrenergic drugs cause dose-dependent bronchodilation when given as aerosols from metered dose inhalers, and a combination of the two can provide better results. If the response to a single agent is unsatisfactory, use of higher doses is advised and the use of a combination of anticholinergic agents and beta-agonists is recommended. With regard to inhaled corticosteroids, a high dose of inhaled beclomethasone dipropionate (1,500 micrograms per day) can be a effective as oral corticosteroids. Step-by-step pharmacologic therapy with the drugs mentioned above should be used in outpatient management of patients with chronic obstructive pulmonary disease. PMID- 9216190 TI - [Clinical benefit of nutritional assessment and support in patients with chronic obstructive pulmonary disease]. AB - Weight loss is common in patients with chronic obstructive pulmonary disease (COPD). Comprehensive nutritional assessment was conducted in two large groups of patients with COPD who were enrolled in the Respiratory Failure Research Program sponsored by the Japanese Ministry of Health and Welfare and the Kinki COPD Research Group. The incidences of mild malnutrition (%IBW < 90%) were 74% and 62%, respectively. The incidences of hypoalbuminemia were low: 10.0% and 6.5%, respectively. The incidence of imbalance in plasma amino acids, which was defined as an abnormally low BCAA/AAA ratio, was as high as 93% in patients with COPD and chronic respiratory failure. The %IBW was significantly related to the FEV1 and to the DLco/VA. The moderately-malnourished subpopulation was characterized by a greater degree of hyperinflation and hypercapnea: the measured resting energy expenditure (REE) was significantly higher than the values in age-matched healthy controls. REE/REEpred was significantly and inversely related to BCAA/AAA and to Pimax. REE was inversely related to FEV1%. REE in the subgroup with severe hyperinflation was significantly higher than REE in those with milder hyperinflation. Among patients with an FEV1% of less than 50%, mortality tended to be higher in those with lower body weight, and this relationship was stronger in patients with an FEV1% of more than 50%. When patients were given a BCAA enriched enteral formula in addition to their usual diet for 3 months, there was a significant increase in body weight, transferrin level, and Pimax. PMID- 9216192 TI - [Chemotherapy for small-cell lung cancer]. AB - Here we review the current treatments for small-cell lung cancer. Cisplatin and etoposide, combined with concurrent or alternating thoracic irradiation, have been considered to be the standard therapy for patients with limited disease. Dose-intensive weekly chemotherapy and high-dose chemotherapy with autologous stem cell transplantation have failed to increase survival in patients with extensive disease. Promising new drugs such as irinotecan and taxol may improve survival in patients with extensive disease. PMID- 9216194 TI - [Surgical treatment for small cell lung cancer]. AB - Surgical treatment for small cell lung cancer had been largely discontinued until the early 1980s, when a retrospective analysis found a high cure rate in some patients. The Japan Clinical Oncology Group (JCOG) also performed a retrospective analysis of 114 patients with small cell lung cancer who underwent surgical resection from 1981 to 1985. The 5-year survival rate was 47% for those in clinical stage I and 20s% for those in stage II and stage IIIA. Whether chemotherapy should be given before or after surgery is unclear. Howevers, In a recent prospective study patients who underwent local radiotherapy soon after the start of chemotherapy survived longer than did those who underwent local radiotherapy after chemotherapy. A phase II trial of postoperative chemotherapy is now being done by the JCOG. The interim results are promising: the 3-year survival rate is 80% in patients whose disease is classified as pathological stage I. PMID- 9216193 TI - [Chemotherapy for non-small cell lung cancer]. AB - The combination of cisplatin and either a vinca alkaloid or etoposide is one of the most effective chemotherapeutic regimens for non-small cell lung cancer. Meta analyses of clinical trials in which cisplatin-based combination chemotherapy was compared with supportive care in patients with non-resectable non-small cell lung cancer showed that the chemotherapy reduced mortality. In the last decade, new agents, including vinorelbine, edatrexate, paclitaxel, docetaxel, irinotecan, topotecan and gemcitabine, have shown promise in the treatment of non-small cell lung cancer, and new agents combined with platinum compounds have reached the level of phase III testing. Randomized studies in which radiotherapy alone was compared with radiotherapy and cisplatin-based combination chemotherapy in patients with stage III disease showed that combined treatment can prolong survival and improve long-term outcome in patients with locally advanced non small cell lung cancer. Recently, new agents combined with radiotherapy have been tested. Regimens in which platinum compounds are combined with new agents hold promise for more successful treatment of this disease. PMID- 9216195 TI - [Recent advances in radiation therapy for non-small cell lung cancer]. AB - Several technical advances in radiotherapy for non-small cell lung cancer have recently been reported. For early superficial cancer in the hilar region, intracavitary brachytherapy with an Ir-192 source combined with external radiotherapy is very effective; the 5-year survival rate can exceed 85%. For peripheral-type early non-small cell lung cancer the local control rate with conventional external radiotherapy remains about 50%. Heavy-particle therapy with protons or heavy-ion beams is promising. Radiotherapy combined with chemotherapy is routinely used to treat locally advanced non-small cell lung cancer. The optimal combination of these treatments is an important subject for future research. PMID- 9216196 TI - [Laser photodynamic therapy for central-type lung cancer]. AB - Laser endoscopic surgery is recognized as an effective treatment for lung cancer. Attention has increasingly been focused on photodynamic therapy (PDT) with dihermatoporphyrin ethers. The Japanese government approved the use of this therapy in October 1995, and reimbursement through the national health insurance system began in April 1996. Over the past decade, 140 patients (283 lesions) with central type lung cancers have been treated at our hospital. Overall complete remission was obtained in 39.6% of 112 lesions, partial remission in 59.4% and no remission in 1.0% The indications for PDT are as follows: 1. Early stage lung cancer curative; among 95 early stage lesions complete remission was obtained in 79 (83.2%), and 71 patients were disease free at 3 to 176 months. 2. Advanced lesions opening of bronchi; overall, "effective" opening of bronchi was achieved in 61 of 81 lesions (75%), in the PDT group, and in 143 of 177 (81%) in the Nd YAG laser therapy groups. 3. Preoperative laser irradiation to increase operability and reduce the extent of operation; the extent of resectin was reduced, or inoperable lesions were made operable in 21 of 24 patients treated. 4. Multiple primary lung cancer. The success of clinical trials of PDT for treatment of lung cancers bodes well for its future use. More stable, definitive, and successful treatment will become possible with the use of new dyes that distribute more evenly in tumor tissue, and with the deeper tissue penetration made possible by longer-wavelength beams. PMID- 9216197 TI - [A new strategy for treating small cell lung cancer]. AB - Recent results from molecular biology have shown that lung cancer is characterized by multiple, sequentially appearing molecular changes that include genetic and epigenetic alterations. Among all types of lung cancer, small cell lung cancer (SCLC) is associated with the lowest rate of 5-year survival. In this symposium, we introduce our findings regarding the c-kit oncogenes in SCLC. We found that the c-kit gene is strongly expressed in SCLC. The c-kit gene was not expressed in normal bronchial epithelial cells, which indicates that this gene is abberantly transcribed in SCLC. In addition, c-kit-positive cases of SCLC showed autophosphorylation in response to recombinant human stem cell factor. Furthermore, adding rh stem cell factor of SCLC cell lines induced a significant chemotactic response and moderate in vitro cell growth. These results strongly suggest that abnormal expression of the c-kit gene may be involved in the pathogenesis of SCLC by autocrine/paracrine stimulation via the c-kit/SCF signal pathway. To overcome drug resistance, we assessed the efficacy of a chimeric toxin targeted to c-kit receptors. PMID- 9216199 TI - [The roles of adhesion molecules, cytokines, and chemokines in eosinophil activation during allergic inflammation]. AB - Patients with hypereosinophila have at least two subpopulation of eosinophils: "normodense" and "hypodense". Hypodense eosinophils can be distinguished by their increased expression of various membrane receptors including IL-5 receptors (J Exp Med 172: 1347) and by the expression of particular proteins (J Immunol 142: 4416). Recently, adhesion molecules have also been found to play an important role in the inflammatory processes in allergic disease. 1) Adhesion molecules were found to be strongly expressed on eosinophils from patients with asthma. 2) Platelet activating factor and induced the expression of adhesion molecules as did the supernatant of mononuclear cells from mite-allergic patients with asthma stimulated either with mite allergen or with a combination of recombinant IL-3, GM-CSF, and IL-5 (Immunol, Lett. 42: 25, '94, & 46: 241, '95). 3) Patients with bronchial asthma had a high level of soluble ICAM-1) (Lancet. 343: 1108, '94). Moreover, the presence of a large variety of membrane receptors and the identification of cytotoxic molecules (mainly granule basic proteins) indicate that eosinophils should be considered effector cells. Therefore the release of granule proteins in response to ICAM-1 and its ligands was studied. The concentrations of eosinophil cationic protein and eosinophil-derived neurotoxin in supernatants of eosinophils were significantly greater in the presence of recombinant soluble ICAM-1 than in its absence (p < 0.05). These results suggest that signals from ICAM-1 and its ligands induce eosinophil activation and are involved in degranulation of eosinophil granule proteins. In addition, reactive oxygen species generated by eosinophils have also been considered capable of causing airway injury at sites of inflammation. The effect of recombinant soluble ICAM-1 and its ligands on eosinophil-induced radical oxygen products was studied. Recombinant soluble ICAM-1 augmented eosinophil oxidative metabolism. Therefore, signaling via adhesion molecules might play an important role in the pathogenesis of allergic inflammation. Specifically, it may activate eosinophils and increase oxidative metabolism or cause degranulation of eosinophil granule proteins. PMID- 9216200 TI - [Rhinovirus infection and expression of adhesion molecules in human tracheal epithelium]. AB - Rhinovirus infection has attracted attention because it can lead to acute exacerbations of chronic inflammatory airway diseases such as bronchial asthma and chronic bronchitis. We established a culture system and inoculated human rhinovirus to human tracheal epithelial cells, and found that infection was augmented by up-regulation of intercellular adhesion molecule-1, which is the receptor for this virus. We also found that human airway epithelial cells infected with rhinovirus were susceptible to a chemical oxidant (H2O2) released by inflammatory cells, which would contribute to acute exacerbations of inflammatory airway diseases. Finally, we found that anti-ICAM-1 antibodies or dexamethasone can inhibit the infectivity to rhinovirus by suppressing ICAM-1, and diminish susceptibility to oxidants in the cultured human tracheal epithelium. PMID- 9216202 TI - [Interactions between bronchial epithelial cells and extracellular matrix proteins]. AB - Attachment and migration of bronchial epithelial cells are important in re epithelialization after tissue injury. We hypothesized that inflammatory cytokines alter bronchial epithelial cell attachment and migration. To test this hypothesis, we studied the effects of mononuclear-cell-conditioned medium on attachment and migration of bronchial epithelial cells in response to fibronectin in vitro. This medium was prepared from bovine blood mononuclear cells that were stimulated with concanavalin A; it stimulated bronchial epithelial cell migration but inhibited attachment to fibronectin. Sephadex G-75 column chromatography of the medium revealed two peaks of activity for stimulation of migration. Activity in the higher molecular weight peak was partially inhibited by anti-TNF-alpha antibodies. Activity in the low-molecular-weight peak was lipid-extractable, which suggests that it reflected an arachidonate metabolite. We also studied the effect of bovine herpes virus-1 infection on migration of bronchial epithelial cells. Infection with this virus reduced the migration of bronchial epithelial cells; by 6 hours after infection, staining of alpha v beta 3 integrins had become more diffuse and was not localized. Thus, mononuclear cells produce inflammatory cytokines with important effects on the migration of bronchial epithelial cells. Viral infection affects the interactions of bronchial epithelial cells with the extracellular matrix. PMID- 9216201 TI - [Role of oxidants in adhesion molecule expression and cytokine production]. AB - Reactive oxygen intermediates such as hydrogen peroxide play an important role in the pathophysiology of acute lung injury, not only as terminal effectors, but also as second messengers in signal transduction; we studied their role in adhesion molecule expression and cytokine production. N-acetylcysteine, an antioxidant, decreased the TNF alpha-induced expression of intercellular adhesion molecule-1 on cultured epithelial cells from human bronchi (BEAS-2A), and inhibited IL-8 production by those cells. In vivo, N-acetylcysteine attenuated the sequestration of polymorphonuclear neutrophils in rat lungs caused by intratracheal lipopolysaccharide. These findings suggest that adhesion molecule expression and cytokine production in the lung are mediated by the production of reactive oxygen intermediates. Because adhesion molecules and cytokines play a crucial role in the pathophysiology of neutrophil-mediated acute lung injury, the inhibition of adhesion molecule expression and cytokine production with anti oxidants such as N-acetylcysteine may be a useful therapeutic strategy. PMID- 9216203 TI - [Role of adhesion molecules in an animal model of endotoxin-induced acute lung injury]. AB - We studied the role of adhesion molecules in acute lung injury caused by lipopolysaccharide (LPS). Forty-eight female guinea pigs were divided into three groups: saline (n = 12); B464, an LPS antagonist, (0.2 mg/kg i.v.) (n = 12); LPS (0.02 mg/kg i.v.) (n = 12); and LPS + B464 (n = 12). The numbers of polymorphonuclear cells (PMN) in blood sampled over 4 hours were counted. Accumulation of PMNs in the lungs was determined by counting the number of PMNs in lung-tissue samples fixed for light-microscopic examination. The lung wet-to dry weight ratio and the 125I-albumin tissue-to-plasma ratio were used to assess lung injury. Human umbilical-vein endothelial cells were treated with B464 and then stimulated with either LPS or tumor necrosis factor. Expression of ICAM-1 and ELAM-1 was estimated by enzyme-linked immunosorbent assay. After LPS injection, light microscopy revealed decreases in peripheral PMN counts, and accumulation of PMNs in the lungs. Increases in the two indices of lung injury were also observed. These changes were attenuated by prior treatment with B464. The LPS-induced increases in ICAM-1 and ELAM-1 expression were dose-dependently suppressed by B464. These results suggest that pulmonary accumulation of activated PMNs plays an important role in LPS-induced lung injury. PMID- 9216205 TI - [Video-assisted bedside pleuroscopy under local anesthesia: use of a rigid cystoureteroscope in patients with undiagnosed pleural effusion]. AB - Up to 20% of pleural effusions remain undiagnosed despite history-taking, physical examination, thoracentesis, and percutaneous closed pleural biopsy. The next diagnostic procedure used is often thoracoscopy under general anesthesia in an operating room. We report a technique for beside pleuroscopy and pleural biopsy that can be done without assistance of surgeons. We performed video assisted pleuroscopy with a rigid cysto-ureteroscope in seven patients with pleural effusion that remained undiagnosed despite extensive clinical evaluation. A sterile 19.8 Fr. rigid cysto-ureteroscope was placed into the pleural space under local anesthesia. Pneumothorax was induced to enhance visualization of the surfaces. Forceps-biopsy specimens were taken of suspicious lesions on the parietal pleural. In three patients the pleural surface appeared smooth and in two the parietal pleural surface was studded. A localized coin-like lesion was seen in one patient, and extensive fibrinogenic adhesions and diffuse opacity of the parietal pleura was seen in another. Using this bedside procedure, we diagnosed pleural tuberculosis in three patients and pleural metastases of adenocarcinoma in one. When done under local anesthesia with a rigid cyst ureteroscopy, video-assisted pleuroscopy can be a safe and useful diagnostic aid in patients with undiagnosed pleural effusion. PMID- 9216204 TI - Neutrophil emigration in the lungs. AB - Neutrophil emigration into the lung occurs in response to inflammatory mediators in the interstitium and the airspace. Emigration through the pulmonary microvasculature occurs through two pathways, one that requires CD11/CD18 and ICAM-1 and one that does not: Which pathway is utilized is determined by the stimulus. The ability of a stimulus to upregulate ICAM-1 by inducing the production of pro-inflammatory cytokines including TNF-alpha appears to determine the selection of the CD11/CD18, ICAM-1, ICAM-1-dependent pathway Recently, a third pathway has been identified that requires CD11/CD18 but not ICAM-1. The ligand for this pathway, as well as the ligands for CD11/CD18, ICAM-1-independent adhesion have not been identified. During recurrent pneumonia, the adhesion molecules required for emigration are different than those utilize during acute inflammation in previously normal lung tissue. For example, studies investigating the role of CD11/CD18 in recurrent pneumonia induced by P. aeruginosa, a stimulus which elicits CD11/CD18-dependent emigration initially, showed that when the organisms are instilled at the same site 7 days after the initial instillation, most emigration occurs through CD11/CD18-independent mechanisms. These studies suggest that when an acute stimulus is placed at a site of resolving inflammation, new pathways of adhesion are recruited. Whether these molecules are the same ones mediating acute CD11/CD18-independent adhesion remains to be determined. In summary, neutrophil emigration in the lung can occur through several adhesion pathways, which pathway is utilized can change during the inflammatory process, and cytokines participate in the selection of the pathway. PMID- 9216206 TI - [Thoracoscopic surgery for diagnosis and treatment of pleural and mediastinal disease]. AB - We studied the usefulness of thoracoscopic surgery in the diagnosis and treatment of pleural and mediastinal lesions. 1. Thoracoscopic surgery was the best method for biopsy of pleural lesions or effusion-retaining disease. Thoracoscopic surgery allowed extensive intrathoracic inspection and collection of relatively large tissue samples, even though it is less invasive than thoracotomy. 2. Thoracoscopic biopsy was useful in the diagnosis of tumorous lesions of the anterior, middle, and posterior parts of the mediastinum. 3. Thoracoscopic surgery is indicated for treatment of the following five pleural diseases: (1) benign pleural tumors; (2) localized mesothelioma; (3) persistent pleural effusions; (4) pyothorax; and (5) chylothorax. 4. Benign mediastinal tumor is a clear indication for thoracoscopic surgery, but utmost caution must be exercised in applying this technique to the treatment of malignant tumors. 5. Thoracoscopic extended thymectomy with a collar incision of the neck is a useful and relatively non-invasive technique for the treatment of myasthenia gravis without thymoma. PMID- 9216207 TI - [Utility of thoracoscopy for diagnosis of pulmonary diseases in clinical pulmonary medicine]. AB - Thoracoscopy is useful for diagnosis of a number of lung diseases. We report our recent experience with thoracoscopy in 86 patients and with video-assisted thoracic surgery in 105 patients. In 70 patients with pleural effusion, thoracoscopic pleural biopsy was done under local anesthesia. All malignant pleural effusions that were not diagnosed by cytologic examination of pleural effusion fluid were diagnosed by thoracoscopy. Especially in malignant mesothelioma, thoracoscopy yielded correct diagnoses. No complication was observed. In 45 patients with a solitary pulmonary nodular lesion and in 27 patients with diffuse interstitial lung diseases, thoracoscopic lung biopsy was done with an "endo-stapler", under general anesthesia. The diagnostic accuracy of this procedure was excellent. Plasma neutrophil elastase and IL-6 levels were measured after surgery as markers of the invasiveness of the procedure. The results indicated that thoracoscopic lung biopsy is relatively safe and non invasive. Thoracoscopy has become an indispensable tool in the daily practice of pulmonology. PMID- 9216208 TI - [Diagnosis and treatment of pulmonary nodules by video-assisted thoracoscopic surgery--surgeons' view]. AB - We reviewed 48 cases of pulmonary nodules in which video-assisted thoracoscopic surgery was done at Jichi Medical School Hospital from 1992 through 1995. The pulmonary nodules comprised 14 malignant tumors (9 lung cancers and 5 pulmonary metastases), 10 benign tumors (7 hamartomas, 2 localized mesotheliomas and 1 tumorlet), 19 granulomas (8 inactive infectious tumors, 7 active infectious tumors, and 4 granulomas as sequelae of other diseases), 4 intrapulmonary lymph nodes, and 1 pulmonary cyst. Conventional operations for lung cancer were done in 7 cases, and 6 were found to be ST-I. Tumor resection by video-assisted thoracoscopic surgery allowed diagnosis of rare diseases and treatment of benign lung tumors and of lung metastases. We conclude this procedure is very useful for diagnosis and treatment of indeterminate pulmonary nodules. PMID- 9216209 TI - [Relief of dyspnea after lung volume reduction in patients with pulmonary emphysema]. AB - We studied dyspnea and pulmonary function during treadmill exercise in 10 patients with pulmonary emphysema before and at least 3 months after thoracoscopic volume reduction. Dyspnea was significantly less after surgery. Ventilatory function, gas exchange, respiratory muscle function, and exercise performance also improved significantly. The degree of volume reduction, estimated by the decrease in FRC as measured by body plethysmography correlated with the degree to which dyspnea was relieved. Volume reduction may be appropriate as a treatment for dyspnea in patients with pulmonary emphysema. PMID- 9216210 TI - [Indications and techniques for surgical treatment of pulmonary emphysema]. AB - We compared two surgical treatments for pulmonary emphysema: unilateral thoracoscopic surgery and bilateral volume reduction (pneumectomy). There were no significant complications with either technique. Symptoms were relieved and pulmonary function improved with both. Post-operative pain was more severe and postoperative blood loss was greater after pneumectomy, but the improvement in pulmonary function was also greater. Possible advantages of bilateral thoracoscopic surgery over pneumectomy should be studied. PMID- 9216211 TI - [Role of alveolar macrophages in the pathogenesis of idiopathic interstitial pneumonia]. AB - To clarify role of alveolar macrophages in the pathogenesis of idiopathic interstitial pneumonia (IIP), we studied (1) the localization and expression of monocyte chemoattractant protein-1 (MCP-1) in IIP by immunohistochemistry and in situ hybridization. (2) the role of MCP-1 in macrophage recruitment to be lung, and (3) the clinical usefulness of measuring MCP-1. (1) In IIP, MCP-1 was observed in cuboidal and flattened metaplastic epithelial cells, alveolar macrophages, and vascular endothelial cells. In contrast, no epithelial cells from patients without IIP stained positive for MCP-1, although alveolar macrophages and vascular endothelial cells were labeled. MCP-1 production by epithelial cells in IIP may be caused by the metaplastic nature of the epithelial cells and may be a main cause of the irreversible progression of IIP. (2) MCP-1 levels in bronchoalveolar lavage fluid (BALF) were significantly higher in the IIP, the interstitial pneumonia due to collagen vascular disease (IP-CVD) and sarcoidosis groups than in normal controls. In the IIP group, the MCP-1 level was significantly higher than in any other patient groups. In all three groups of patients, the monocyte chemotactic activity in BALF correlated positively with the MCP-1 levels in BALF, and were neutralized by anti-MCP-1. (3) BALF MCP-1 levels were significantly higher than serum MCP-1 levels in the IIP group, and they were lower in the IP-CVD and non-specific interstitial pneumonia groups. Serum MCP-1 levels reflected the activity of interstitial lung diseases, especially during treatment with corticosteroids. These results indicate (1) that MCP-1 plays a significant role in the recruitment of monocytes into the lung in IIP, (2) that measuring MCP-1 levels both in BALF and in serum may help discriminate IIP from other types of interstitial lung diseases, and (3) that monitoring the serum MCP-1 level may be useful in estimating the activity of interstitial lung diseases. PMID- 9216212 TI - [Clinical significance of serum surfactant proteins A and D in idiopathic interstitial pneumonia]. AB - Pulmonary surfactant is synthesized and secreted by alveolar type II cells. The major components of surfactant are phospholipids and four distinct surfactant specific proteins: SP-A, SP-B, SP-C, and SP-D. The concentrations of SP-D and SP A were measured in sera of patients with idiopathic interstitial pneumonia (IIP), by enzyme-linked immunosorbent assay with monoclonal antibodies against human SP D and SP-A. The concentrations of SP-D and SP-A in samples from the patients were, respectively, 5.7 and 2.8 times higher than those in samples from healthy volunteers; 86.2% and 71.4% of the patients had concentrations of SP-D and SP-A, respectively, that were more than 2 standard deviations greater than the mean of the control values. Moreover, the serum SP-D and SP-A concentrations appeared to reflect the disease activity of IIP. There was a negative significant correlation between the concentrations of SP-A in serum and in bronchoalveolar lavage fluid. These results suggest that SP-D and SP-A, which are primarily secreted from alveolar type II cells into the alveolar lumen, can enter the bloodstream easily due to injury at the alveolar-capillary membrane. We conclude that measurement of SP-D and SP-A in sera can provide an easily identifiable and useful clinical marker for the diagnosis of IIP, and can be used to predict the disease activity of IIP. PMID- 9216213 TI - [Clinical usefulness of KL-6 as a serum marker of idiopathic interstitial pneumonia]. AB - KL-6 is a high-molecular-weight glycoprotein that is classified in cluster 9 among pulmonary cell antigens. Levels of KL-6 were found to be abnormally high in sera from patients with various types of interstitial pneumonitis such as idiopathic interstitial pneumonia, which can progress to pulmonary fibrosis. Levels of KL-6 in serum can be useful serum markers in the diagnosis of idiopathic interstitial pneumonia, in monitoring in the evaluation of disease activity, as a prognostic factor, and as a predictor of response to therapy. PMID- 9216214 TI - [Effects of in vivo instillation of genes coding for cytokines on pulmonary fibrosis]. AB - Interstitial pneumonia is characterized by alveolitis that results in interstitial fibrosis. To study the role of humoral factors in the pathogenesis of interstitial fibrosis, we introduced expression vectors into Wistar rats via the trachea, to cause local overexpression of these humoral factors in the lung. Genes for human interleukin (IL)-6 and for the IL-6 receptor caused lymphocytic alveolitis without marked proliferation of fibroblasts. In contrast, overexpression of the genes for human transforming growth factor (TGF)-beta 1 and for human platelet-derived growth factor (PDGF)-B caused only mild cellular infiltration in the alveoli. However, both caused marked proliferation of fibroblasts and deposition of collagen fibrils. Introducing an expression vector that coded for a mutant form of the PDGF beta receptor that lacks its cytoplasmic domain markedly alleviated the pathohistologic changes caused by bleomycin in murine lungs. These findings show that TGF- and PDGF-B may be closely related to fibrosis in the lung, and that artificial regulation of them may be effective for treatment of lung fibrosis. PMID- 9216215 TI - [How can we monitor and treat patients with acute exacerbations of idiopathic interstitial pneumonia]. AB - Levels of KL-6 in serum were measured in 17 patients during acute exacerbations of idiopathic interstitial pneumonia (IIP). The patients were divided into three groups: "effectively" treated (n = 4), "ineffectively" treated (n = 4), and "untreated" with steroid pulse therapy (n = 9). In the "ineffective" group, the level of KL-6 in serum was significantly higher than that in the "effective" group. These results suggest that KL-6 is useful for assessing patients with acute exacerbations of IIP. In a separate study ONO-5046, a specific neutrophil elastase inhibitor, was given in 40 institutions to find out if it is useful in treating IIP. The PaO2/FIO2 ratio after the administration of ONO-5046 was significantly improved in high-dose group, which suggests that this agent is useful in treating patients with IIP. PMID- 9216216 TI - [Idiopathic interstitial pneumonia: profiles of the subsets, and prognosis]. AB - Idiopathic interstitial pneumonia (IIP) comprises diseases with different clinical courses and different histopathologic characteristics. Clinical differential diagnosis and differentiation based on histopathologic characteristics reflecting inflammatory lung injuries and fibrosis are used to classify the pneumonia and to decide on management. Corticosteroids and immunosuppressants are given, but knowing the correct indications, timing, and duration of therapy depends on knowledge of the natural course of the diseases and on long-term observation of the clinical courses. We compared the clinical profiles, prognoses, and histopathologic characteristics of subsets of idiopathic interstitial pneumonia, and therapeutic approaches based on their pathophysiological differences. PMID- 9216217 TI - [New strategies for the etiologic study of interstitial pneumonia: familial registration and sib-pair analysis]. AB - Etiology of idiopathic interstitial pneumonia (IIP) remains unknown. While many studies focused on the analysis of individual patients, available information is limited for the further study. Having experienced a case, 3 of 5 brothers suffered from IIP and 2 of them complicated with lung cancer, intensive familial history taking with chest photos and functional information for fibrotic lung was performed. Among 37 families in which at least 2 members were examined, 14 families (37.8%) had minimum 2 members with fibrotic lungs. Unexpectedly high familial accumulation of fibrotic lung indicates the possibility of multi-gene involvement for the pathogenesis. We need to start group studies to register sib pairs with fibrotic lungs and systematize the automated genetic analysis for the responsible genes. PMID- 9216218 TI - [Vitamin K deficiency syndrome caused by antituberculous agents]. AB - Vitamin K deficiency caused by antituberculous agents was examined in clinical patients and in experimental rats. When antituberculous agents given to the patients who had only total elental diet because of small intestinal dysfunction, a marked increase in plasma PIVKA-II and a decrease in thrombo-test value were observed. These changes were quickly normalized by administration of vitamin K, despite of succeeding or stopping of antituberculous agents. In rat experiments, effects of four agents (ethambutol, isoniazid, paraaminosalicylate, and rifampicin) on prothrombin time were studied. Among these agents, only rifampicin prolonged prothrombin time. This prolongation depended on drug doses and duration of administration. In addition to this hypoprothrombinemia, an increase in plasma PIVKA-II was also observed, and these changes were normalized within 24 hours of vitamin K administration. These data suggest that rifampicin inhibits vitamin K epoxide reductase interfering re-use of vitamin K and caused vitamin K deficiency in patients with total elental diet. PMID- 9216219 TI - [A case of giant aberrant pancreas with cyst formation]. PMID- 9216220 TI - [A case of gastric, pulmonary, and submandibular gland mucosa-associated lymphoid tissue lymphomas with colon and submandibular gland amyloidosis]. PMID- 9216221 TI - [Torsion of an accessory spleen]. PMID- 9216222 TI - [Long-term survival cases of hepatocellular carcinoma with portal tumor thrombus by repeated arterial infusion chemotherapy using implantable port system]. PMID- 9216223 TI - [Autopsy case of a 22-year-old female with hepatic failure due to alcoholic liver cirrhosis]. PMID- 9216225 TI - [A case of disseminated intravascular coagulation (DIC) during interferon-alpha treatment of chronic hepatitis C]. PMID- 9216224 TI - [Two cases of Basedow disease complicated by acute hepatitis]. PMID- 9216226 TI - [A case of acute hepatitis E]. PMID- 9216228 TI - [A case of peritoneum malignant mesothelioma responding to arterial infusion chemotherapy]. PMID- 9216227 TI - [A case of synchronous triple early cancer occurring in the stomach, colon and gallbladder]. PMID- 9216229 TI - [Human senescence, cancer and telomerase]. PMID- 9216231 TI - [Molecular mechanisms of self-incompatibility in Brassica]. PMID- 9216230 TI - [Chemical modification of histones during cell growth and differentiation]. PMID- 9216232 TI - [World smallest motor, ATP synthase]. PMID- 9216233 TI - [A mammalian telomerase component gene TLP1]. PMID- 9216235 TI - [Chemokines and HIV-1 second receptors]. PMID- 9216236 TI - Cloning of Dolly stirs hornet's nest. PMID- 9216234 TI - [Immunosuppressive substances induced by liver grafting]. PMID- 9216237 TI - Poor handling of mammography. PMID- 9216239 TI - Thalamic stimulation: an innovative treatment for tremor. PMID- 9216238 TI - Pediatrics and the Nashville General Hospital--Part I. PMID- 9216240 TI - Lymphogranuloma venereum vs. incarcerated inguinal hernia: the Gordian knot unraveled. AB - A 33-year-old woman presented with a two-day history of left inguinal pain and a palpable "knot." She was suspected of having an incarcerated inguinal hernia but instead was found to have lymphogranuloma venereum. The diagnosis is typically made from a history and physical examination, but may be supported by serologic complement fixation studies or by testing of lesion aspirates using immunofluorescent antibody testing. PMID- 9216243 TI - A new approach to nutrition education. PMID- 9216241 TI - Regional and ethnic differences in stroke in the southeastern United States population. PMID- 9216242 TI - Traumatic pseudoaneurysm of the superficial temporal artery. PMID- 9216245 TI - Opioid peptides derived from hemoglobin: hemorphins. AB - Investigation of hemoglobin peptic hydrolysate has revealed the presence of biologically active peptides with affinity for opioid receptors. Two peptides, VV hemorphin-7 and LVV-hemorphin-7, were resolved by a combination of size exclusion and reversed phase HPLC. A new spectroscopic method based on the second order derivative spectra analysis of aromatic amino acids has been developed. This method allows qualitative and quantitative evaluation of hemorphins generated by peptic hemoglobin hydrolysis. Using this method, a kinetic study of hemorphins appearance has been undertaken. In this paper, we also evidenced the generation of VV-hemorphin-7 from globin by peritoneal macrophages. In regard to this result, the putative physiological role of hemorphins is discussed. PMID- 9216244 TI - Indinavir-induced nephropathy. PMID- 9216246 TI - Milk protein-derived opioid receptor ligands. AB - Milk is mammalian characteristic and is of particular importance for humans: Mother's milk or its substitutes from cows' milk are absolutely essential nutriments for the neonate and cows' milk also represents a basic foodstuff for adults. However, in addition to their well-known nutritive role, milk constituents apparently are also able to carry specific information from the milk producer's to the milk receiver's organism: Thus, a number of milk protein fragments has been shown to behave like opioid receptor ligands able to address opioidergic systems in the adult's or in the neonate's organism. With respect to the proteins, which they are derived off these peptides have been named alpha casein exorphins or casoxin D (alpha-casein), beta-casomorphins or beta-casorphin (beta-casein), casoxin or casoxin A, B, or C (k-casein), alpha-lactorphins (alpha lactalbumin), beta-lactorphin (beta-lactoglobulin) or lactoferroxins (lactoferrin). Only casoxins and lactoferroxins display antagonistic properties; the other peptides behave like opioid receptor agonists. Most of the information available so far has been collected about beta-casomorphins. These peptides obviously can be released from beta-casein in the adult's or in the neonate's organism, where they might elicit opioid effects in the frame of a regulatory role as "food hormones". Several synthetic beta-casomorphin derivatives have been shown to be highly specific and potent mu-type opioid receptor ligands which frequently have been used as standard tools in opioid research. PMID- 9216247 TI - Biochemical properties of regulatory peptides derived from milk proteins. AB - Biologically active peptides derived from milk proteins are inactive within the sequence of the precursor proteins but can be released by enzymatic proteolysis. Based on structure-activity studies, peptides with a defined bioactivity show common structural features. Moreover, many milk protein-derived peptides reveal multifunctional bioactivities. Bioactive peptide fragments originating from milk proteins should be taken into account as potential modulators of various regulatory processes in the body. Opioid peptides are opioid receptor ligands with agonistic or antagonistic activities. Angiotensin converting enzyme (ACE) inhibitory peptides can exert an antihypertensive effect. Immunomodulating casein peptides have been found to stimulate the proliferation of human lymphocytes and the phagocytic activities of macrophages. Antimicrobial peptides have been shown to kill sensitive microorganisms. Antithrombotic peptides inhibit the fibrinogen binding to a specific receptor region on the platelet surface and also inhibit aggregation of platelets. Casein phosphopeptides can form soluble organophosphate salts and may function as carriers for different minerals, especially calcium. In relation to their mode of action, bioactive peptides may reach target sites (e.g., receptors, enzymes) at the luminal side of the intestinal tract or after absorption, in peripheral organs. The physiological significance of bioactive peptides as exogenous regulatory substances is not yet fully understood. Nevertheless, several bioactive peptides derived from milk proteins have been shown to exert beneficial physiological effects. Milk-derived peptides were already produced on an industrial scale and as a consequence these peptides have been considered for application both as dietary supplements in "functional foods" and as drugs. PMID- 9216248 TI - Antihypertensive peptides derived from food proteins. AB - This paper reviews the angiotensin I converting enzyme inhibitory peptides originated from food materials and enzymatic hydrolysate of different kinds of proteins. Focus was put on the peptides derived from milk casein by the action of the proteolytic system of lactic acid bacteria. Some of the peptides exhibit significant antihypertensive effects in spontaneously hypertensive rats. Some new topics relating to these antihypertensive peptides are introduced. The possible significance of bioactive peptides derived from food in vivo is also discussed. PMID- 9216250 TI - Bone marrow immunoregulatory peptides (myelopeptides): isolation, structure, and functional activity. AB - Myelopeptides (MPs) are bioregulatory mediators of bone marrow origin. Several individual MPs have been isolated from the supernatant of porcine bone marrow cell culture by successive solid phase extraction and reversed-phase high performance liquid chromatography. Two of them, MP-1 (Phe-Leu-Gly-Phe-Pro-Thr) and MP-2 (Leu-Val-Val-Tyr-Pro-Trp), were synthesized and their biological activities were comprehensively studied. Both hexapeptides display pronounced immunoregulatory activity but their final effects as well as mechanisms of action are different. Peptides MP-1 and MP-2 are identical to conservative fragments 33 38 alpha- and 31-36 beta-chains of hemoglobin, respectively. The sequences of other isolated MPs have no homology with any functional protein. The role of MPs in bioregulatory processes in vivo is discussed. PMID- 9216249 TI - Primary structure and biological activity of hemoglobin-related hypothalamic peptides. AB - Five individual fractions from bovine hypothalamic extract, displaying coronary constrictory activity, were isolated and sequenced. All of them belong to the hemorphin group of hemoglobin-derived peptides. These peptides bind calmodulin and activate calmodulin-dependent enzymes. The relationship of isolated peptides with other members of the hemorphin group is discussed. Several new fragments of hemoglobin alpha- and beta-chains with yet unidentified activity were obtained from the same source. Their amino acid sequences have considerable overlap with the known sequences of hemoglobin fragments isolated from other tissues. PMID- 9216251 TI - The hemorphins: a new class of opioid peptides derived from the blood protein hemoglobin. AB - Hemorphins are endogenous peptides belonging to the family of "nonclassical" or "atypical" opioid peptides. They are generated by enzymatic hydrolysis of the beta-, kappa-, delta-, or epsilon-chain of the blood protein hemoglobin. Originally, the hemorphins were isolated from enzymatically treated bovine blood. In recent years hemorphin structures have been identified as naturally occurring peptides in brain, plasma, and cerebrospinal fluid. This article will review recent studies of the hemorphins regarding their structures, mechanisms for their release, and their biological actions. A particular emphasis will be directed to their role in exercising human and their clinical relevance. PMID- 9216252 TI - Profiling of endogenous peptides as a tool for studying development and neurological disease. AB - The use of a method to follow changes in endogenous peptide production, as they occur in biological studies, is an excellent complement to other molecular techniques. It has the unique ability to characterize peptides that have been produced from protein precursors, and instrumentation is available that provides high resolution peptide separations that are quantitative, sensitive, and amenable to automation. All tissues express a large number of peptide species that can be visualized, or profiled, on chromatographic separations using reverse phase high-performance liquid chromatography. This large number of peptides offers many potential molecules that can be used to identify biological mechanisms associated with experimental paradigms. Peptide analysis has been used successfully in many types of studies. In this review, we outline our experience in using peptides as biological markers and provide a description of the evolution of peptide profiling in our laboratories. Peptide expression has been used in studies ranging from how brain regions develop to identifying changes in disease processes including Alzheimer's disease and models of stroke. Some of the findings provided by these studies have been new pathways of peptide processing and the identification of accelerated proteolysis on proteins such as hemoglobin as a function of Alzheimer's disease and brain insult. Peptide profiling has also proven to be an excellent technique for studying many well-known nervous system proteins including calmodulin, PEP-19, myelin basic protein, cytoskeletal proteins, and others. It is the purpose of this review to describe our experience using the technique and to highlight improvements that have added to the power of the approach. Peptide analysis and the expansion in the instrumentation that can detect peptides will no doubt make these types of studies a powerful addition to our molecular armamentarium for conducting biological studies. PMID- 9216253 TI - Hemoglobin as a source of endogenous bioactive peptides: the concept of tissue specific peptide pool. AB - Scattered literature data on biologically active hemoglobin-derived peptides are collected in the form of tables. Respective structure-functional correlations are analyzed and the general conclusion is reached that hemoglobin fragments must have a profound physiological function. Evidence is presented that generation of hemoglobin fragments starts inside the erythrocytes. At that stage alpha- and beta-globin chains of hemoglobin predominantly give rise to relatively long peptides containing ca. 30 amino acid residues. The primary proteolysis is followed by the next degradation step coupled with excretion of newly formed shorter peptides form red blood cells. Both the primary and the secondary proteolysis products are subjected to further stepwise C- and N-terminal chain shortening, giving rise to families of closely related peptides that are actually found in animal tissue extracts. The possible sites of primary proteolysis are compared with the positions of the exposed secondary structure elements within the monomeric alpha- and beta-globins as well as the tetrameric hemoglobin. Two tentative schemes are proposed for hemoglobin degradation, one of which starts at the globin loops exposed on the surface of the tetramer and the other, at monomeric globins where more sites are available for the action of proteases. The concept of a "tissue-specific peptide pool" is formulated, describing a novel system of peptidergic regulation, complementary to the conventional hormonal and neuromodulatory systems. According to that description, hemoglobin is only a single example, although an important one, of a vast number of functional proteins providing their proteolytically derived fragments for maintaining the tissue homeostasis. PMID- 9216254 TI - Characteristics of winter and summer aerosol mass and light extinction on the Colorado plateau. AB - This paper focuses on the spatial variability of fine mass and extinction budgets taking data from the winter and summer months of 1992. The study area included southern California, southern Nevada, southern Utah, Arizona, and parts of New Mexico. Two types of monitoring sites were operated: intensive and secondary or satellite. At the intensive sites, all major aerosol species were measured as well as extinction or scattering. At the satellite sites, trace elements including sulfur and hydrogen, absorption, and gravimetric fine mass were measured. Where all aerosol species are measured, the spatial variability of extinction budgets is examined assuming an externally mixed aerosol. At the satellite sites, an approximated fine mass budget is derived and the variability of these budgets in space and time are examined. This effort was part of a study called Project MOHAVE (Measurement of Haze and Visual Effects) carried out with the principal objective of understanding the relative contribution of regional and local sources to visibility impairment on the Colorado Plateau and specifically, the Grand Canyon. Generally, the contribution of sulfates, organics, and absorption to extinction are about equal at 20-30% with the coarse mass fraction being about 10-20%. The one exception is in southern California where the nitrate contribution tends to be higher in the winter than summer. During the summer, concentration gradients tend to be spread out across the study area, while during the winter months, variability in concentration and budgets tends to occur on a smaller scale. PMID- 9216255 TI - Human visual sensitivity to plumes with a Gaussian luminance distribution: experiments to develop an empirical probability of detection model. AB - This paper discusses results of a research project designed to develop an empirical model that could be used as a tool to predict human visual sensitivity to plumes. The resultant probability of detection algorithm (PROBDET) allows one to estimate the probability of a plume of known size, shape and contrast being detected visually. As a basis for the algorithm, a series of laboratory experiments using a high threshold signal detection procedure and computer generated images of plumes with Gaussian luminance distributions was conducted to measure human visual sensitivity to plumes. Results of the laboratory experiments are compared with results of contrast sensitivity experiments that examined visual sensitivity to stimuli with square and sine wave luminance distributions. An example of the PROBDET algorithm is presented to demonstrate its potential usefulness for assessing how probability of detection estimates change as plume size and contrast parameters vary. Since this research was designed to build on existing knowledge, a discussion of that knowledge and how it relates to the research conducted is also presented. The focus of this discussion is on the human visual system (HVS) and on how visual sensitivity is affected by factors such as the luminance of the stimulus and the surround, the luminance distribution of the stimulus, the size of the surround, and the size and spatial frequency characteristics of the stimulus. PMID- 9216256 TI - Organochlorine insecticide residues in African Fauna: 1971-1995. AB - Organochlorine insecticides (OCLs), which were introduced in the decade following World War II, were used extensively in Europe, the U.S., and other developed countries into the 1970s. However, data began to accumulate on their persistence in soils and aquatic sediments, their potential to be taken up into animal tissues and to bioconcentrate in birds and mammals in the higher tropic levels of food chains and even in humans. As a result, registration authorities phased out their use progressively, in Europe and the U.S., from 1973 onward. However, the production of OCLs in developed countries and their use in developing countries continued through the 1970s and 1980s into the 1990s because they were, no longer under patent agreement, were inexpensive to manufacture, and were very effective in pest control. In Africa, the use of OCLs continued well into the 1990s for the control of mosquitoes, tsetse flies, and desert locusts as well as to combat various crop, animal, and human pests. Some of these uses involved extensive spraying of large areas of nonagricultural land, thereby exposing many groups and species of wildlife to their residues. Although there is some evidence of a gradual decline in the use of OCLs in Africa, they are still being used in appreciable quantities. During the past 25 yr, there have been 50 published reports of OCL residues in the various groups of invertebrate and vertebrate animals constituting the African fauna. These have been based on a diverse range of surveys, target animals, sampling methods, and analytical techniques. Moreover, they are extremely regionally-biased, the most intense surveys being in Zimbabwe, Kenya, Egypt, and South Africa. DDT was the most commonly used OCL, accounting for about half the total use, followed closely by dieldrin and HCH. Birds and fish have been sampled most intensively, with relatively few studies on other taxa. We reviewed the OCL residue data on African fauna from these reports and summarized the maximum and mean residues in the various groups of terrestrial and aquatic invertebrates and vertebrates. Overall, residues in the fauna were the greatest for DDT, followed in turn by those of dieldrin, HCH, endosulfan, and endrin, with small amounts of aldrin and toxaphene being found in some animals. There were relatively few reports of OCL residues in terrestrial invertebrates and virtually none in aquatic invertebrates. Only a few reports demonstrated OCL residues in terrestrial vertebrates, although high levels of DDT, dieldrin, and HCH were found in crocodile eggs and large residues of dieldrin occurred in bats, squirrels, and monkeys. Considerable OCL residues were reported in a few species of fish, especially Barbus, Clarias, Hydrocynus, Labeo, Sarotherodon, Epiplatys, and Synodontis. These residues were at levels that could have caused chronic toxicity or behavioral changes. The calculated maximum and mean OCL residues in the various elements of the African fauna until 1995 were compared with those calculated for corresponding faunal groups in Europe and the U.S. from their development and introduction up to 1973. The OCL residues reported in African fauna between 1971 and 1975 tended to be significantly higher overall than those published for Europe and the U.S. In particular, residues of DDT and dieldrin in African birds and their eggs were greater than those that had been incriminated as causing significant eggshell thinning and reproductive failure in European and U.S. aquatic and terrestrial birds up to 1973. Additionally, high DDT and dieldrin residues were reported from some species of African fish at levels that could potentially affect their reproduction, have chronic toxic and behavioral effects, and even drastically affect populations. Holistic case studies on the use of OCLs to control tsetse flies and desert locusts were discussed. OCL levels in trophic levels of fauna associated with Lake Kariba (between Zambia and Zimbabwe) were summarized. (AB PMID- 9216257 TI - Rubber tire leachates in the aquatic environment. AB - Tires have a deleterious effect on the environment. This review discusses the background of scrap tires discarded in the environment, including tire composition, adverse environmental effects, threats to public health and safety, and solid waste management. Despite the widespread use of scrap tires in environmental applications, both land-based and aquatic, data on the indicators of environmental degradation are extremely scarce. Indicators of environmental degradation include analysis of chemicals within the water and sediment, analysis of contaminants within organisms, and analysis of the biological effects of these compounds on plants, animals, microbes, and organelles. Although these indicators are most useful when used in parallel, a review of the available information on chemical characterization of tire leachate from tire storage facilities, manufacturing, usage in recycling applications, and toxicity exposure studies, of vegetation surveys from waste tire areas and reviews of mammalian tire product toxicity, and of toxicity, mutagenicity, and carcinogenicity of tire exposure in experimental aquatic animals, microbes, and organelles is presented. The major characteristics of these studies are discussed in specific sections. The "Discussion and Conclusions" section discusses and summarizes the biological effects and chemical characterization of tire leachates. A global environmental perspective is included to improve our understanding of the deficiency of the current knowledge of tire leachate toxicity from various sources and to encourage interdisciplinary studies to establish the pattern of pollution associated with waste tire management. PMID- 9216258 TI - Factors affecting atrazine fate in north central U.S. soils. AB - Atrazine persistence and fate are influenced by many factors, the interactions of which are difficult to predict. Several models, such as LEACHP (Wagenet and Hutson 1989), have been used as tools to estimate losses and identify variables that will impact the magnitude of loss. The LEACHP model was evaluated for predicting atrazine movement in sandy loam, silt loam, and clay loam soils during three consecutive years (two dry and one wet) in Minnesota (Khakural et al. 1995). Considering the broad range in soil properties and climatic conditions used in testing, the model performed well. However, these are only estimates, and additional field studies need to be conducted to verify model results. In a report by Fausey et al. (1995), the amount of atrazine found in groundwater throughout the Midwestern region was reported to be much below the MCL. However, specific sites in the Midwest may struggle with atrazine problems from both point and nonpoint sources of contamination. Some states, such as South Dakota, have created groundwater protection areas that alert growers and the public to sensitive areas where contamination may occur because of soil type, depth to groundwater, and distance to public wellheads. Wisconsin has developed a tiered managerial strategy, or zoning approach, in which restrictions are matched to pollution detections (Wolf and Nowak 1996). The USEPA has mandates for states to implement generic management plans to prevent pesticide contamination of groundwater. Chemical-specific plans by states will be required for at least five pesticides, one of which will be atrazine. Best management practices have been and are continuing to be developed to aid the grower in lessening the adverse impacts of atrazine. With continuing research into understanding the problem and developing solutions, and with adaptation of these recommendations by growers, the use of effective, inexpensive herbicides may continue with minimal off-site environmental effects. PMID- 9216259 TI - Home care for venous leg ulcers. AB - More than a quarter of the adult population in the United States is afflicted with lower extremity venous insufficiency, and 1 in 100 have had, or now have, stasis ulcers. Most of these patients will be treated on an outpatient basis, with many of them requiring home health care. The cost to treat venous ulcers alone has been estimated at $750 million to $1 billion a year. Understanding the pathogenesis and treatment of the problem is imperative as home health care nurses move into an era of cost containment and demographic shift toward an increasingly larger elderly population. PMID- 9216260 TI - Unlicensed assistive personnel and lay caregivers in the home. AB - As a result of the changes in the health care delivery system, professional home health agency staff must provide clinical assessments, care planning, and patient outcome evaluation to higher acuity patients with limited resources. Because the number of home visits per patient is closely monitored by managed care companies, duplication of services may not be reimbursed, causing financial loss to home care agencies. Every aspect of care from admission to discharge must be coordinated to ensure that each patient receives the best care available within the limitation of the number of reimbursable or capitated visits. Professionals on an interdisciplinary team must know the legal scope of practice as well as the competencies of all members of the team. Decisions relating to delegation of patient care tasks to UAP and to lay caregivers are complex. Professional staff must consider the patient's condition, the caregiving environment, and the amount of social support available for the patient and family. The knowledge, skills, and abilities of the caregivers must then be matched to the needs of the patient. Because on-site supervision in the home setting is limited, nurses and therapists are required to know how to safely delegate patient care tasks and supervise others. PMID- 9216261 TI - Serving cultural communities in home health. PMID- 9216262 TI - Health care consumer education: alternative teaching methods. AB - Anything that can be handled, manipulated, or related to previous life experiences leaves a lasting impression with the health care consumer and therefore is remembered longer. The nurse also must consider the literacy level, ethnic background, and indigenous language of the consumer when choosing the educational technique. Everyone responds quickly to scenarios to which they can relate, in which their personal perception is explored, and in which the language and logic of the educational experience make sense and are consistent with the consumer's lifestyle. PMID- 9216264 TI - Heart attack: what you don't know can hurt! AB - Do belching, chills, fatigue, and pressure just above the ribs sound like the symptoms of a heart attack? To most patients, they probably don't. However, research is indicating that a victim's actual symptoms often do not match his or her expectations of what a heart attack should feel like. As a result, many sufferers do not seek medical attention, or they delay it, which can result in permanent damage to the heart muscle or even death. A recent research study indicated that most patients with recent heart attack (74%) interviewed had symptoms different from what they expected. As a result, medical treatment was significantly delayed. PMID- 9216263 TI - Home health nursing practice. AB - Home care practice demands role clarity and definition. To this end, the American Nurses Association addressed these standards in the Standards of Home Health Nursing Practice. These standards and the listed responsibilities should be a part of nursing orientation to home care practice and operations. In the ever changing health care environment, which has created multifaceted nursing roles unique to home care, these standards can become the framework for effective home care nursing activities. PMID- 9216265 TI - The role of pharmacists in home care. PMID- 9216266 TI - Designing an infection control program. AB - Designing and implementing an infection control program for home care poses many challenges. Some of these challenges stem from a long-standing lack of research and on-site expertise in the home care setting. Recent changes in health care and the proliferation of external regulations regarding home care practice further complicate the development of an effective and efficient infection control program. PMID- 9216267 TI - Snap judgment. What is it? A superficial phlebitis in a collateral vein. PMID- 9216269 TI - The world is your community. PMID- 9216268 TI - Patient Self-Determination Act: advocating for the patient's wishes. AB - The United States has a long tradition of protecting an individual's rights. A recent example is the Patient Self-Determination Act (PSDA), federal law (1992, P.L. 101-508). It specifically allows competent individuals to communicate their preferences for life-sustaining medical treatment before they become incapacitated and are unable to make them known. It requires health care agencies, including home health agencies, receiving funds through Medicaid or Medicare programs to provide information and education about advance directives (ADs) to their patients, staff, and the community. Home health care agencies need to have protocols in place to be in compliance with the law. Agencies have to inform patients about their rights to make health care decisions and must maintain written policies about the implementation of those rights. The federal law prohibits home care providers from making the patient's care conditional on the existence of an AD or in any way altering the quality of care provided because of his or her wishes. PMID- 9216270 TI - American Academy of Home Care Physicians. Are you ready for this? PMID- 9216272 TI - Stand and deliver! PMID- 9216271 TI - On accreditation of healthcare organizations. Determining eligibility for Accreditation of Home Pharmaceutical Services. AB - Many misperceptions exist in the health care community about the Joint Commission's accreditation of home pharmaceutical services, primarily related to when pharmacy services are considered home care and which patients or services are eligible for survey. This article concentrates on eligibility for survey and accreditation of just one of the six eligible home care services-home pharmaceutical services. The concept of home care and pharmaceutical services is difficult for most people to understand. Unlike other home care services, pharmacy services cannot be provided directly in the patient's home. Pharmacy laws prohibit pharmacists from compounding and dispensing directly in the patient's home. The pharmacists must do these activities in a licensed pharmacy. Also, most patients who receive medications from pharmacies take their own medications in their homes. So the differences between home care pharmacy and ambulatory or retail pharmacy are much less clear-cut than the delivery of more traditional home care services such as nursing or home health aide services. PMID- 9216273 TI - Caring ... for ourselves. PMID- 9216274 TI - Nurses should take action to avoid occupational latex allergy. PMID- 9216275 TI - Latex allergy. PMID- 9216276 TI - Pepper spray antidote successful in one emergency department. PMID- 9216277 TI - Latex allergy in children. PMID- 9216278 TI - Varicella-zoster virus pneumonia in the adult patient. PMID- 9216280 TI - The emergency nurse practitioner: an educational model. PMID- 9216281 TI - HIV occupationally exposed health care workers and prophylactic drug therapy. PMID- 9216279 TI - The effect of administered crystalloid fluid temperature on aural temperature of moderately and severely injured children. AB - OBJECTIVE: Warm intravenous fluid (W-IVF) administration is the standard of care to prevent hypothermia in injured adults. It is argued that such administration may not be helpful for treating injured children, because children often do not require as much intravenous fluid (i.v.f.) as adults. The purpose of this study was to compare the effects of W-i.v.f. to room temperature intravenous fluid (RT i.v.f.) administration on aural temperature (Ta) in injured children during the first hour of trauma resuscitation. DESIGN: A randomized, controlled repeated measures trial. SETTING: Emergency department, intensive care unit, and diagnostic areas in a level I pediatric trauma center. SAMPLE: Thirty moderately or severely injured children, ranging in age from 2 to 17 years (mean age = 8.9 years; SD = 4.4). METHODS: Eligible children were randomly assigned to receive either W-i.v.f. or RT-i.v.f. on ED arrival. Warmed IVF was administered with the Hotline fluid warmer (SIMS Level 1, Rockland, Mass). Aural temperatures were measured on arrival and every 10 minutes for 1 hour with a Core-Check Tympanic Thermometer (IVAC Medical Systems, San Diego, Calif). The level of significance for hypothesis testing was set at 0.05 (two-tailed). RESULTS: Groups were comparable in age, gender, weight, amount of infused i.v.f., Revised Trauma Score, room temperature, and baseline Ta. On average, Ta for the W-i.v.f. group increased by 0.25 degree C from baseline to final Ta, whereas Ta for the RT i.v.f. group decreased by 0.32 degree C from baseline to final Ta. Repeated measures analysis of covariance, treating baseline Ta as a covariate, demonstrated that Ta response profiles were similar (p = 0.06). CONCLUSIONS: When comparing the changes between baseline and final Ta for the W-i.v.f. and RT i.v.f. groups, the standardized difference in temperature change was 0.62. Although results of the repeated measures analysis of covariance were not statistically significant, the standardized difference in temperature changes was large enough to warrant administration of W-i.v.f., even at slow flow rates, to prevent hypothermia in injured children. PMID- 9216282 TI - Malaria in the emergency department. AB - Malaria can a be serious, life-threatening illness. Data suggest that there are thousands of cases in the United States each year and that infection rates are increasing. Initial patient presentation will often be at the ED triage desk; patients usually have flulike symptoms and a fever. A high index of suspicion for individuals at risk of malaria clarified by a brief but thorough travel history facilitates early identification of presumptive cases and prompt, effective care. Early antibiotic treatment and aggressive supportive care are the mainstays of treatment. Serious P. falciparum infection should always be presumed and treated until proven otherwise. PMID- 9216284 TI - Latex products in the hospital environment. PMID- 9216283 TI - Severe latex allergy: three first-person accounts. PMID- 9216285 TI - Latex allergy: policy, procedure, and latex-safe box. PMID- 9216286 TI - Response to ski lift disaster by rural Canadian clinic. PMID- 9216287 TI - The tapestry of care. PMID- 9216288 TI - Conscious sedation roundtable: a collaborative practice model for problem solving in the emergency department. PMID- 9216289 TI - DNA evidence collection. PMID- 9216290 TI - Our new infrared emergency department. PMID- 9216291 TI - Latex allergy sufferers--the clock is ticking. PMID- 9216292 TI - Workers' compensation for illness and injury. PMID- 9216294 TI - Teaching the teacher: first-semester notes of a new undergraduate nursing instructor. PMID- 9216293 TI - Workers' compensation and latex allergy: dos and don'ts. PMID- 9216295 TI - Mylar or bust. PMID- 9216296 TI - Testing a test. PMID- 9216297 TI - SANE program locations: pros and cons. Sexual Assault Nurse Examiner. PMID- 9216298 TI - Strategies for improving trauma documentation. PMID- 9216299 TI - A 62-year-old man with jaw pain and history of angina. PMID- 9216300 TI - Creating a latex-safe environment: Riddle Memorial Hospital's response to protect patients and employees. PMID- 9216301 TI - Born again. PMID- 9216302 TI - The ice capades. PMID- 9216303 TI - Bedside. PMID- 9216304 TI - ELASTIC and ALERT: help for nurses and others with latex allergies, help for those who work with latex. Education for Latex Allergy/Support Team and Information Coalition. Allergy to Latex Education and Resource Team. PMID- 9216305 TI - [Touch--a measure of tact]. PMID- 9216306 TI - [Unavoidable invasion into the patient's intimate sphere, no reason for shame]. PMID- 9216307 TI - [Latex allergy. When protective gloves make you ill]. PMID- 9216308 TI - [Health care in Havanna. Attention instead of aspirin]. PMID- 9216309 TI - [Every day is different]. PMID- 9216310 TI - [Discharge planning should always be part of it]. PMID- 9216311 TI - [Treatment of lower extremity arteriopathies. Stimulation of the spinal cord]. PMID- 9216312 TI - [Considerations on vomiting. Understanding for better care]. PMID- 9216313 TI - [Measurement of hemoglobin Alc in diabetic children. Combining education and play]. PMID- 9216314 TI - [Nursing story. The olive branch]. PMID- 9216315 TI - [Knotty aspects and strategies of response: the evolution process of nursing theory]. PMID- 9216316 TI - Diagnostic techniques in cardiac disorders: Part I. AB - Advances in technology over the past 15 years have given birth to a new field of multidisciplinary cardiovascular care and research, that of perinatal cardiology. Today's neonatal nurse must become familiar with the battery of tools that may be enlisted to assist in the assessment, diagnosis, intervention, and evaluation of cardiac disorders and therapeutic maneuvers in the neonate. In this first part of a two-part article, the history and physical examination, the chest x-ray, and the electrocardiogram are reviewed. The second part will discuss the role of both invasive and noninvasive "high-tech" diagnostic techniques in the management of cardiac disorders. PMID- 9216317 TI - Review of cerebral palsy, Part II: Identification and intervention. AB - Cerebral palsy (CP) can be identified and classified by thorough evaluation using multiple assessment techniques. Clinical signs, symptoms, associated disorders, and methods used to evaluate developmental disorders have been described. Because some of the previously used evaluation methods have not been adequate tools, new ways of measuring outcome have been proposed. Multidisciplinary treatment protocols have been recommended. Simplistic plans of care will not be appropriate because of the complexity of the disorder(s) and the unique characteristics of the individual and family. Therefore, thoughtful and individualized plans of care that may include multiple surgical and nonsurgical interventions must be developed for each child with CP. In addition, more data are needed, using new evaluation techniques, to determine the efficacy of early intervention. PMID- 9216318 TI - Maternal recall of the neonatal intensive care unit. AB - This study examined how mothers of prematurely born three-year-old children retrospectively recall their responses to their infant's hospitalization in the neonatal intensive care unit (NICU). Forty-four mothers of three-year-old prematurely born children were interviewed as part of a longitudinal study. Data from maternal interviews were analyzed using the analytic inductive method. Findings support the hypotheses that were based on the Parental Stress in the ICU model. Three years after the birth of their premature infants, mothers reported vivid memories of stress related to the appearance and behavior of their infants, the pain and procedures the infants endured, alterations in their role as parents, and stress related to the infant's illness severity and uncertainty about infant outcomes. Prenatal problems, such as high-risk pregnancy or birth, infant loss, and disturbances in family support, were also recalled as sources of stress. Findings have implications for family-centered nursing care in NICUs. PMID- 9216319 TI - An alternative approach for neonatal clinical pathways. AB - Defining a generic plan of care for neonatal intensive care patients seems like an impossible task. Many institutions have elected to purchase plans already developed by other practitioners. Implementing these plans can be a frustrating and discouraging experience. In our organization, administration supported multidisciplinary development of pathways, enabling us to review current practice and recommend enhancements in care provision that maintained quality while reducing resource utilization. This article describes how the staff in our 45 bed, Level III NICU, staffed with attending neonatologists, approached the challenge of defining care practices for a diverse patient population. PMID- 9216320 TI - A case history: apnea in the newborn. PMID- 9216321 TI - In our unit: skin care for VLBW infants. PMID- 9216322 TI - Hot issues in 1997. PMID- 9216323 TI - Welfare reform tracking--impact on children. PMID- 9216324 TI - Comfort measures and analgesia. PMID- 9216325 TI - Educational consortiums for NICU staff. AB - Remember that forced change can provide opportunities for creative endeavors. Take the initiative to contact your colleagues and identify potential collaborative projects. The temporary alliances formed for an educational consortium may develop into other exciting joint ventures. PMID- 9216326 TI - Syndrome of inappropriate antidiuretic hormone. PMID- 9216328 TI - The image of nurses as "caring angels". PMID- 9216327 TI - What are the legal rights of the unborn child? PMID- 9216329 TI - The waiting game. PMID- 9216330 TI - Our woman in Westminster. PMID- 9216331 TI - Turn down the heat. PMID- 9216332 TI - Yes, in our backyard. PMID- 9216333 TI - Don't go changin'. PMID- 9216334 TI - Cash concerns. PMID- 9216335 TI - Think 'care'. PMID- 9216336 TI - Mobile carers. PMID- 9216337 TI - The quality of mercy. PMID- 9216338 TI - Learning curve. PMID- 9216339 TI - The wonder drug. PMID- 9216340 TI - In celebration of nursing skills. PMID- 9216341 TI - Pressure picture. PMID- 9216342 TI - Broadcast. PMID- 9216343 TI - Research network for child health nursing. AB - A new network for RCN members interested in child health or children's nursing research is due to be launched at the end of June. The Research in Child Health (RiCH) network is part of the RCN Society of Paediatric Nursing and aims to promote research and research-based nursing in the specialty. Network co ordinator Sarah Neill explains how interested nurses can become involved. PMID- 9216344 TI - Polish and British nurses' responses to patient need. AB - This research study explored the differences between the responses of a cohort of British nurses and a cohort of Polish nurses to a patient's expression of need. Using a vignette to elicit the responses, the study showed that the cohorts gave similar responses in terms of cheering the patient up, offering explanation and showing warmth. Differences were demonstrated in relation to collecting information, giving advice and suggestions, and referring the patient to the doctor. Both cohorts, however, rarely gave responses which demonstrated empathy. The authors conclude that better training in therapeutic communication is needed. PMID- 9216345 TI - Influencing decision making. PMID- 9216346 TI - Establishing a hospital-based pressure care service. AB - The emphasis on pressure area care in all nursing specialties and the need to reduce actively the incidence of pressure sores, means that pressure areas must be monitored from the time the patient arrives in hospital to the time of discharge and afterwards. The use of effective pressure-relieving equipment plays a major part in the prevention of pressure sores (Hitch 1995). This paper examines the way Salisbury Health Care NHS Trust tackled this problem in hospitals and, in doing so, established an in-house pressure care service. PMID- 9216347 TI - Alternating pressure systems: reducing user error. AB - Pressure sores remain a major clinical challenge for nearly every sector of health care. The Department of Health has recognised this and in the 1994/95 NHS Priorities and Planning Guidance encouraged healthcare providers to set annual targets for the reduction of the prevalence of sores by 5 per cent. It has been argued that this is not straightforward (NHS CRD 1995) especially as many areas do not monitor the problem and so do not know how effective they are at preventing sores. Reduction cannot be attempted unless an effective prevention plan is in place. The causes of pressure sores are multifactorial, so the prevention strategies must reflect this. PMID- 9216348 TI - Pressure sore pilot scheme. PMID- 9216349 TI - The relevance of cultural diversity to patient education. PMID- 9216350 TI - Readability of patient information booklets for women with breast cancer. AB - Providing accurate information, in both verbal and written formats, is seen as an important component of patient care. For individuals diagnosed with cancer, acquiring information may be a particularly pertinent issue in terms of coping with the disease. Numerous information booklets are available for people with cancer which aim to provide information on various aspects of care and treatment. This British study examined the readability of 50 information booklets available to women with breast cancer using the SMOG and Flesch reading tests. Generally the information booklets were found to have a high reading age, arguably not suitable for the majority of the United Kingdom (UK) population. This study has implications for health care professionals who provide written information as a supplement or substitute for verbal information. PMID- 9216351 TI - Personalities and alimentary behaviors in obese patients. AB - The actual tendency in the care of obese patients is the association of dietetic information with an eating behavior therapy. Studies attempting to attribute the origin of obesity to psychiatric pathologies are contradictory. We studied whether certain eating disorders are more specific to a personality type. We studied eating disorders with the Eating Disorder Inventory (EDI) test in 281 obese women compared to 252 age-matched non-obese women. Both obese patients and non-obese volunteers were divided into four groups depending upon their personality (PERSONA test). This test defines four types of personality, based on the level of emotion (expansive or reserved) and the degree of power (dominant or consenting). According to our study, eating disorders vary between the four personality groups and were significantly higher in the facilitating group (consenting and expansive) compared to the three other obese groups. Neither promoting (expansive and dominant) nor controlling obese patients (dominant and reserved) present eating disorders. The analyzing obese patients (reserved and consenting) are reticent when it comes to consulting (18%) since they distrust others. Analyzing obese patients present an interpersonal distrust and an interoceptive awareness. The group which presents most eating disorders is that of facilitating obese patients (consenting and expansive). These present eating disorders of the compulsive types favored by interoceptive awareness, body dissatisfaction, ineffectiveness, and maturity fears. The diversity, even the absence, of eating disorders brought to evidence by our tests based upon different personality types should allow better understanding of the psychological and behavioral causes of weight gain and the means for improving compliance in the following of an obese patient. PMID- 9216352 TI - Incorporation of cognitive-behavioral treatment into the medical care of chronic low back patients: a controlled randomized study in German pain treatment centers. AB - Cognitive behavioral treatment has been incorporated into standard medical treatment procedures in German pain centers. Acceptance of the treatment by patients and outcome in terms of pain, coping, and disability was investigated. Components of the psychological treatment are education, relaxation and imagery, modifying thoughts and feelings, enhancement of pleasant activities, and training of good postural habits. The program was conducted in a group setting in accordance with a treatment manual and consists of 12 weekly 2.5-h sessions. A two-factor experiment with repeated measures on one factor was applied. Ninety four consecutive patients with low-back pain were randomly assigned to an experimental group having a combined medical and cognitive-behavioral treatment, or to a control group with medical treatment only. Assessments were taken pre treatment, post-treatment, and--in the treated group only--at a 6-months follow up. At each assessment, patients kept a pain diary over a period of 4 weeks, and filled in self-report questionnaires. The sample consisted of 36 experimental and 40 control subjects at post-treatment. Experimental subjects reported less pain, better control over pain, more pleasurable activities and feelings, less avoidance and less catastrophizing. In addition, disability was reduced in terms of social roles, physical functions and mental performance. The results were maintained at follow-up. Patients who only received medical treatment showed little improvement. Data indicate that the program meets the needs of the patients and should be continued. PMID- 9216353 TI - Promotion of healthy lifestyle: knowledge and attitudes of primary care physicians. AB - This study was conducted in Riyadh city to determine the knowledge and attitudes of primary health care physicians regarding promotion of certain healthy lifestyles. Using a stratification technique, 50% of the male physicians were selected at random. A validated self-administered questionnaire was distributed to (and recollected from) the participants. One physician refused to participate resulting in a response rate approaching 99%. Most of the physicians (77.5%) were shown to have satisfactory knowledge, while only 20% possessed reasonably positive attitudes. Physicians above 40 years of age were more knowledgeable than younger age group (P < 0.0001). There was a general pessimism about compliance of patients to lifestyle advice. The relatively acceptable knowledge possessed by physicians was not properly translated into positive attitudes towards promotion of healthy lifestyle. To improve the situation, a greater emphasis should be placed on health promotion during undergraduate and postgraduate training of physicians. PMID- 9216354 TI - Euthanatics: implementation of a protocol to standardise euthanatics among pharmacists and GPs. AB - The purpose of this study was to evaluate the implementation of a protocol to standardise euthanatics among pharmacists and general practitioners (GPs). Data over 1993 and 1994 were collected by means of an anonymous postal questionnaire sent to all pharmacists (n = 37) and all GPs (n = 283) working in the area in which the protocol was implemented. In total, 76% of the pharmacists and 63% of the GPs responded. All pharmacists and 65% of the GPs were aware of the existence of the protocol and all pharmacists and 42% of the GPs were also familiar with the content of the protocol. Both pharmacists and GPs had fairly positive attitudes towards the importance and possibility of the standardisation of euthanatics. Of the GPs who performed euthanasia or assisted with suicide during the research period, 59% made use of one or more standard packages. The majority of pharmacists and GPs were satisfied with the standard packages, and all GPs indicated that they intend to use the packages again in the future. This study shows that the implementation of standardised euthanatics was quite successful. PMID- 9216355 TI - Literacy assessment in a cardiovascular nutrition education setting. AB - We assessed functional literacy of hypercholesterolemic or hypertensive African Americans (n = 339) prior to their participation in a nutrition education program. A word pronunciation and recognition test using 20 common cardiovascular or nutrition terms was first developed based on correlations with standardized reading achievement test scores, then administered to program participants. Nearly half (48%) had word recognition scores equivalent to a < or = 8th grade reading level. Lower scores were associated with less education, lower income, unemployment, heavier work activity if employed, less healthy diets, history of heart disease or diabetes, and higher depression scores (all P < 0.01); several of these associations were independent of education. The educational materials were geared to a 5th to 8th grade reading level. However, when both audiotaped and printed instruction were provided, individuals with reading scores < or = 8th grade preferentially used the tapes. This brief and relatively unobtrusive literacy assessment may help to identify persons who can benefit most from audiovisual approaches to cardiovascular nutrition education. PMID- 9216356 TI - Patient education for the millennium: beyond control and emancipation? AB - This paper reports on a study carried out in Nottinghamshire, UK, which focused upon patient education for low back pain in general (family) practice. This study found that patient education could not be viewed simply as either a repressive social control mechanism or as a vehicle for patient empowerment and social change. The paper suggests that a new theoretical and practical orientation to patient education is required which transcends binary categorizations. The existing control/empowerment dichotomy offers a persuasive, yet restrictive conceptualization of patient education which has created rival camps of theoreticians and practitioners intent on demonizing each other. In the light of presented findings, the study suggests a new trajectory for patient education focusing on local possibilities for change. PMID- 9216357 TI - Freire revisited: old truths in new disguise. A reaction to Skelton's 'Patient education for the millennium'. PMID- 9216359 TI - Improved medical understanding of human health. AB - The time has come to examine aspects of health and disease on a broader basis than immediate cause and effect. Recognition of the biological processes of adaptation shaped by natural selection has become a requirement for improved understanding of human health in the modern environment, and the time has come for its incorporation into medical teaching and clinical practice. PMID- 9216360 TI - The use of problem-based learning in dealing with cultural minority groups. AB - Minority peoples like the Romanies have divergent cultures. Typical cultural aspects for medical personnel to consider would include greetings and other communication, family and social support, dressing and habits of cleanliness, marriage and sexuality, honor, and other issues of importance to any human being. Some minority cultures have no geographic boundaries but they still may adopt the lifestyles of the country they live in. Physicians have to reckon with these different cultural patterns when dealing with patients. Patients must be treated equally at the same time when their personal needs require individual consideration. This consideration is reflected in both verbal and non-verbal communication with the other. Both the sender and the receiver of a message would need to know of the other. Minority groups tend to know more about the majority groups than vice versa. Most health care providers belong to the majority group and would be expected to learn more about the other. Problem-based learning can help students to understand attitudes of minority patients (like the Romanies) and handle the situation. In this instance, the students collected theory base from existing legal, cultural, and other resources and interviewed a Romany woman to verify that the information pertaining to the female case was correct. This combination of theory and experience was considered useful in preparing a case presented to a seminar with 116 medical and dental students in 1994. PMID- 9216361 TI - [Developing a language for nursing. Interview with Patricia Benner]. AB - This (shortened) interview from 1995 contains Patricia Benner's thoughts on the relationship between nursing practice, language and nursing science. Benner alludes to the philosophical and theoretical sources of her work and uses a narrative in order to show her method of interpretation. PMID- 9216362 TI - [Nursing as an art in practice and science]. AB - This paper relies on the assumption that care and science are practices in MacIntyre's sense. Every practice can be carried out on different skill levels. Both practices, care and science are intricately linked to each other. Practice on the first three levels described by Dreyfus & Benner is theory driven, while the works of experienced and expert practitioners can be compared to art. Works from care and science are used to illustrate works of art, but also, for comparing and contrasting the competent and expert practices. PMID- 9216363 TI - [Nurse practitioners as counselors]. AB - Giving advice is among the most important tasks of the clinical nurse specialist. In this article, the authors examine various aspects of counselling in order to contribute to the further analysis and development of the specialist's professional agenda. Starting out with a brief discussion of recent changes in nursing, the authors analyse the main activities of the clinical nurse specialist on the basis of their respective areas of expertise. The actual process of counselling is examined against the background of the acquired level of professional counselling skills. The article sums up a thesis completed at the end of a course on organizational development, team counselling, supervision, and coaching under the auspices of the Swiss Association for Applied Psychology (SAAP). PMID- 9216364 TI - [Do clinical nurse practitioners affect the working environment and the quality of care?]. AB - This correlational study examined the influence of a new kind of advanced nurse practitioner in Switzerland with the role of providing complex care, consulting with patients and families, fostering the growth of staff, enhancing team functioning and contributing to nursing knowledge through research. Based on systems theory the perception of staff about their work environment and their motivation to engage in continuing education as well as the patients' satisfaction with care and trust in the nurses were measured outcomes. Instruments were constructed for this study, tested and standardized. Data from three hospital units with an advanced practitioner were compared to three units without a practitioner similar in size, staffing, patient acuity and system of nursing care. Staff reported better guidelines for care, more frequent feedback and better professional advice on units with an advanced practitioner while the quality of communication, professional stimulation and support with difficult patients was comparable on all units. Motivation to further one's education was generally high. In units with an advanced practitioner, there was increased interest in psychosocial topics, and the availability of such literature was better. Patients were highly satisfied in all units, but there was a slight difference in patient trust. It was somewhat easier for patients on units with an advanced practitioner to ask the nurses questions. The study suggested that the advanced practitioner should contribute most by strengthening team cohesion through constructive feedback as well as enhancing a climate for continuing learning through better resources, guided case discussions and opportunities for shared problem solving. PMID- 9216365 TI - [The concept of nursing as a basis for the practice of care. 1]. AB - The understanding of nursing can be seen as a basis for practising care and as a potential for its evolution. This survey aims at analysing registered nurses understanding of care. Using a qualitative approach seventeen nurses were interviewed. The author used 50 picture cards as a help for the interviewees to express their ideas in a narrative interview. Using a semi-structured interview the ideas were examined in more depth. The findings are described in the second part of this article. PMID- 9216366 TI - [Perspectives of nursing science. Formation of a theory in a practice discipline]. AB - In this article some problems about nursing theories are discussed. It is presumed that a science does not turn into a nursing science because it deals with nursing practice. Crucial is the way in which the practice is viewed, i.e. the question which theory is the starting point. Even normative nursing theories must be tested out in reality though this verification cannot be made exclusively with the methods of logical empiricism. The obligation for nursing theories to fit the metaparadigm as well as the concept of metaparadigm are discussed. It is also stated that nursing theories do not correspond to the diversity of everyday nursing. In order to create a complex everyday live, grounded theories are needed. For a better understanding, two grounded theories are described. PMID- 9216367 TI - [Ambulatory care in Germany--orientation towards community care?]. AB - In the German healthcare system, two major trends contribute to the growing need for ambulatory care. One is the demographic increase of the older population; the other is the growing preponderance of chronic diseases. The required services often surpass the human resources of the family and ask for professional nursing service. The combination of lay caregiving and professional work is a necessary growing phenomenon that calls for new concepts in home care. In this perspective the idea of the so called "Sozialstation" (social center), which started their work in 1970, is an interesting concept towards more systematic planning and interdisciplinary cooperation and reversing the trend towards hospitalization. This literature-based article looks critically at accomplishments and limitations of Sozialstationen. PMID- 9216368 TI - [Power in the hospital. Considerations on a structural problem]. AB - This article tries to establish elements for an analysis of the power relations in hospital. After an explanation of the notion of power with recurrence to Max Weber, it presents different forms of the exercise of power. The traditional authoritarian form of power is compared with modern forms that are less personal and more anonymous. Then, the article tries to explain the special relationship between power and medicine. The basis for the analysis of this relationship is taken from Michel Foucault's analysis of power who describes medicine as the switch position of modern forms of exercise of power. He defines this under two different points of view: firstly, medicine works as an institution of disciplinary power, secondly, medicine is the sustainer of the bio-power, a kind of power that works by giving life. PMID- 9216369 TI - [An occupation for small hands]. PMID- 9216370 TI - [Osteoporosis: a true problem, but mostly of prevention]. PMID- 9216372 TI - [Ambulatory care and space]. PMID- 9216371 TI - [10 years of emergency forensic medicine at the Hotel Dieu]. PMID- 9216374 TI - [Speaking of ambulatory care facilities in the United States]. PMID- 9216373 TI - [Ambulatory changes ... or new adventures coming from the Far West]. PMID- 9216375 TI - [Political stakes in ambulatory management]. PMID- 9216376 TI - [Economic stakes in ambulatory management]. PMID- 9216377 TI - [Recommendations for a patient before ambulatory surgery]. PMID- 9216378 TI - [How to conceive and organize an ambulatory surgery unit]. PMID- 9216379 TI - [Description of an autonomous ambulatory facility]. PMID- 9216380 TI - [Walking to have hand surgery: explanation of ambulatory surgery]. PMID- 9216382 TI - [The old gentleman]. PMID- 9216381 TI - [Multiple sclerosis, at the heart of medical and social realities]. PMID- 9216383 TI - [Obesity? A question of weight and measures]. PMID- 9216384 TI - [Seen, read and understood ... here and elsewhere. Painting studios at the hospital, a remarkable initiative]. PMID- 9216385 TI - [The authorities are mobilizing against hepatitis C]. PMID- 9216386 TI - [When therapy is leisure]. PMID- 9216387 TI - [Health in prison: what is new?]. PMID- 9216388 TI - [Wet wound debridement, an essential part of wound healing]. PMID- 9216390 TI - [Skin changes: diagnosis and actions]. PMID- 9216389 TI - [The debridement of ulcers: a delicate activity]. PMID- 9216391 TI - [A nursing consultation about chronic wounds in the hospital]. PMID- 9216392 TI - [Under what circumstances does the dressing have to be sterile?]. PMID- 9216393 TI - [Tomorrow's dressings]. PMID- 9216394 TI - [Functional and esthetic scars: an adventure]. PMID- 9216395 TI - [A nutrition program for autistic children]. PMID- 9216396 TI - [I wrote on my uniform]. PMID- 9216397 TI - [Right use of perfume in a nursing service]. PMID- 9216398 TI - [Breaking the isolation...]. PMID- 9216399 TI - [In the course of time ... here and elsewhere]. PMID- 9216400 TI - [AIDS: advantages and disadvantages of double dispensation]. PMID- 9216401 TI - [The hand in ... the glove]. PMID- 9216402 TI - [The church and AIDS]. PMID- 9216403 TI - [A patient's testimony]. PMID- 9216404 TI - [The wasting in AIDS. How to interrupt the cycle]. PMID- 9216405 TI - [The forgotten epidemic. AIDS in the aged]. PMID- 9216406 TI - [AIDS as a disease of body and spirit]. PMID- 9216407 TI - [Palliative care and AIDS]. PMID- 9216409 TI - [Towards a hospital without pain]. PMID- 9216408 TI - [Syphilis and AIDS]. PMID- 9216410 TI - [Pain. Neurophysiological basis]. PMID- 9216411 TI - [Psychological aspects of pain]. PMID- 9216412 TI - [The different types of pain]. PMID- 9216413 TI - [The UPSA Pain Institute]. PMID- 9216414 TI - [Methods of pain evaluation]. PMID- 9216415 TI - [Pain. Pharmacological treatment]. PMID- 9216416 TI - [Pain in children]. PMID- 9216417 TI - [Lies]. PMID- 9216418 TI - [Therapeutic isolation]. PMID- 9216419 TI - [Motivation, indifference and loss of motivation in nurses]. PMID- 9216420 TI - [The oral corticoids]. PMID- 9216421 TI - [Nursing and caring]. PMID- 9216422 TI - [Caring and drug addiction]. PMID- 9216423 TI - [Self-support of drug users. Caring]. PMID- 9216424 TI - [Family groups. Caring for families]. PMID- 9216425 TI - [Caring and drug addicts. Families are mobilizing]. PMID- 9216426 TI - [Physical activity and sports. How an activity can turn into nursing care]. PMID- 9216427 TI - [The social role of the psychiatrist]. PMID- 9216429 TI - [I am timid, but I am taking care of myself]. PMID- 9216430 TI - [At the heart of the reform is the process of contracting]. PMID- 9216431 TI - [From theory to practice]. PMID- 9216432 TI - How the university medical center survived by going "back to the future". PMID- 9216433 TI - Urbs in rure redux: changing risk factors for rural HIV infection. AB - The purpose was to ascertain risk factors for HIV infection in a predominantly rural population using descriptive epidemiologic studies performed at a university health sciences center. Participants included adult patients with HIV infection or AIDS who were cared for between January 1982 and January 1993. The relative frequency of cases in minority and female heterosexual patients increased significantly. The male to female ratio among blacks with HIV infection declined to 1.1:1 during the final 3 years of the study. Patients who believed they had acquired infection in Virginia were more likely to cite a rural area of acquisition and to have had multiple heterosexual partners but were less likely to have had male homosexual contact than patients who believed they had been infected in other states. HIV continued to spread into rural areas of Virginia, and the gender ratio among blacks with HIV declined throughout the study. Having multiple heterosexual partners, the main risk factor for HIV transmission worldwide, may now result in HIV infection in rural Virginia. PMID- 9216434 TI - Evidence of three clinical subgroups in patients with dual atrioventricular nodal pathways. AB - We attempted to test the hypothesis that dual atrioventricular (A-V) nodal pathways with second-degree atrioventricular block (2nd A-V block) present as a different clinical entity from those with A-V nodal reentranttachycardia (AVNRT). By evaluation with Holter monitoring (2.9 +/- 2.5 recordings/patient) and 12-lead electrocardiogram (11.9 +/- 11.6), 177 patients with dual A-V nodal pathways could be divided into three subgroups. Thirty-two patients had 2nd A-V block only (2nd A-V block group), 57 had AVNRT only (AVNRT group), 88 had neither 2nd A-V block nor AVNRT (silent group), and none had 2nd A-V block and AVNRT both. Electrophysiologic studies showed that the atrio-His interval was significantly greater (P < 0.0001) and the maximal 1:1 atrioventricular conduction rate was lower (P < 0.0001) in the 2nd A-V block group than in the other two groups. These differences were nullified after the administration of atropine. These results suggest that patients with dual A-V nodal pathways can be classified into three clinical subgroups based on the presence of either 2nd A-V block or AVNRT. We suggest also that patients of the 2nd A-V block group may have a more augmented vagal tone on the A-V node than the other two groups. PMID- 9216435 TI - Serum albumin level at admission: mortality and clinical outcome in geriatric patients. AB - We evaluated serum albumin at time of admission, within 72 hours, in 135 geriatric patients who were older than 70 years to establish its role as a predictor of death and clinical outcome at time of discharge. Serum albumin values were reduced significantly in patients who died compared with those who were discharged in unchanged/impaired and improved conditions (3.01 +/- 0.68 g/dL, 3.18 +/- 0.55 g/dL, and 3.65 +/- 0.52 g/dL respectively, P < 0.0001). A correlation between serum albumin concentration at admission and number of days elapsed from admission and death was found (r = 0.43, P < 0.05). Mortality rate was 38.6% in patients with serum albumin values < 3.3 g/dL compared with 14.1% in those with albumin values > or = 3.3 g/dL (P < 0.005). Similar results were obtained even when the main diagnostic conditions, such as cardiocerebrovascular disease and cancer, were considered. The results demonstrate that in geriatric patients the serum albumin level at admission may be a predictor of mortality and clinical outcome at discharge. PMID- 9216436 TI - ACE inhibition reduces left ventricular mass independent of pressure without affecting coronary flow and flow reserve in spontaneously hypertensive rats. AB - Systemic and regional (including coronary) hemodynamics were studied in spontaneously hypertensive and normotensive Wistar Kyoto rats after 3 weeks of treatment with one of the three doses of the angiotensin converting enzyme inhibitor, ramipril. The effects of respective treatments on cardiovascular mass and systemic, coronary, and regional hemodynamics (at rest, during maximal treadmill exercise, and during dipyridamole infusion) then were evaluated in conscious rats using radiomicrosphere techniques. Low-dose ramipril (10 micrograms/kg/day by gavage) neither decreased arterial pressure nor reduced cardiac mass. However, medium (100 micrograms/kg/day) and high (1 mg/kg/day) doses reduced total cardiac and left ventricular masses to the same extent in spontaneously hypertensive rats, despite a much greater fall in arterial pressure with a high dose. Resting cardiac index, and myocardial and all other organ blood flows remained unchanged in both strains. When compared with Wistar Kyoto rats, coronary circulation was impaired in untreated spontaneously hypertensive rats (ie, reduced coronary flow and flow reserve and increased minimal coronary vascular resistance during dipyridamole infusion). This remained unchanged by ramipril. Furthermore, significant (and comparable) increases in cardiac index and myocardial blood flow and decreases in coronary vascular resistance were produced by maximal treadmill exercise in both strains. This also was unaffected by ramipril. These data showed that angiotensin converting enzyme inhibition with suboptimal and optimal hypotensive doses of ramipril reversed left ventricular hypertrophy in spontaneously hypertensive rats, but coronary flow, flow reserve, and minimal coronary vascular resistance remained unchanged despite left ventricular hypertrophy reversal. PMID- 9216437 TI - Digoxinlike immunoreactive factor isolated from human pleural fluid is structurally similar to digoxin. AB - To further define the chemical structure of human endogenous digoxinlike immunoreactive factors (DLIF) we used human pleural effusions as a source of the substance. Digoxinlike immunoreactive factor activity was detected by radioimmunoassay in the pleural fluid of each of four patients; average concentration was 0.35 ng/mL. The chemical profile of DLIF was determined by initial extraction and concentration of DLIF by ion exchange chromatography followed by reverse phase-high-pressure liquid chromatography (RP-HPLC) separation and purification. Using high-pressure liquid chromatography cochromatography of DLIF, together with several radioactively marked glycosides, we observed a single peak of DLIF activity that was chromatographically identical to digoxin. The present study further supports the recent finding that DLIF is related structurally to the cardiac glycosides, and for the first time it has been proven that DLIF is present in pleural fluids. PMID- 9216438 TI - Use of glycoprotein IIb/IIIa inhibitors in acute coronary syndromes. PMID- 9216439 TI - The effects of potassium depletion and supplementation on blood pressure: a clinical review. AB - Nonpharmacologic treatment currently is recognized as an important part in the treatment of hypertension, and the role of dietary potassium intake in blood pressure (BP) control is becoming quite evident. Clinical studies have examined the mechanism by which hypokalemia can increase BP and the benefit of a large potassium intake on BP control. Epidemiologic data suggest that potassium intake and BP are correlated inversely. In normotensive subjects, those who are salt sensitive or who have a family history of hypertension appear to benefit most from the hypotensive effects of potassium supplementation. The greatest hypotensive effect of potassium supplementation occurs in patients with severe hypertension. This effect is pronounced with prolonged potassium supplementation. The antihypertensive effect of increased potassium intake appears to be mediated by several factors, which include enhancing natriuresis, modulating baroreflex sensitivity, direct vasodilation, or lowering cardiovascular reactivity to norepinephrine or angiotensin II. Potassium repletion in patients with diuretic induced hypokalemia improves BP control. An increase in potassium intake should be included in the nonpharmacologic management of patients with uncomplicated hypertension. PMID- 9216440 TI - Autoimmune hemolytic anemia, primary adrenal insufficiency, and the antiphospholipid syndrome. AB - Autoimmune hemolytic anemia and adrenal insufficiency are rarely associated with the antiphospholipid antibody syndrome. A 49-year-old woman with a history of deep venous thrombosis and recurrent miscarriages was found to have active autoimmune hemolytic anemia after being admitted to the hospital for cholelithiasis. The patient was treated with corticosteroids and underwent laparoscopic cholecystectomy 1 month later. Two weeks after surgery she had acute adrenal insufficiency. Activated partial thromboplastin time was prolonged, and antiphospholipid antibodies were detected in significant titer. Her illness responded well to corticosteroid therapy. Her direct Coombs' test remained positive. It appears that the antiphospholipid antibody syndrome contributed to the development of venous thrombosis, recurrent miscarriages, autoimmune hemolytic anemia, adrenal insufficiency, and indirectly, pigment stone cholelithiasis in this patient. PMID- 9216441 TI - Spontaneous healing of pancreatic abscess after fistulization to the duodenal bulb. AB - A 70-year-old man was admitted to the hospital because of sudden, upper abdominal and back pain. Laboratory and image data indicated acute pancreatitis. Shortly after the admission, pancreatic and liver abscess with bacteremia developed. Antibiotic therapy seemed effective. A month later, spontaneous fistulization of the pancreatic abscess to the duodenal bulb was found by gastroduodenal fiberscopy. Injection of contrast medium into the duodenal orifice showed that the fistula was draining the abscess and that no other fistula formed from the abscess. Endoscopic retrograde cholangiopancreatogram indicated no fistula formation to the pancreatic duct. The pancreatic abscess became smaller and was not visible using computerized tomography and ultrasonography 3 months later and thereafter. Closure of the duodenal orifice was ascertained by the endoscopy. It is suggested that retrograde infection from the fistula was prevented by the single fistulization to the acidic duodenal bulb, which is not supposed to allow most bacterial growth. Pancreatic abscess usually necessitates operative treatment, even with fistulization to the alimentary tract. It seems likely that the single, small fistulization to the bulb, in addition to the lack of underlying disease and medical and nutritional support, facilitated the spontaneous healing process. PMID- 9216442 TI - Acute massive postoperative pleural effusion associated with asymptomatic hepatitis C-induced cirrhosis of the liver. AB - The development of an acute pleural effusion during the immediate postoperative period after abdominal or pelvic surgery is not uncommon. In symptomatic patients, pleural effusions often are thought to result from a complication of pulmonary embolism or parapneumonic effusion. We present a patient in whom an acute unilateral hepatic hydrothorax developed after elective total abdominal hysterectomy. Pleural effusion continued to reaccumulate for several days. After extensive efforts failed to reveal the cause of effusion, intraperitoneal radioisotope study confirmed a peritoneopleural communication associated with unsuspected and asymptomatic hepatitis C-induced cirrhosis of the liver with portal hypertension. PMID- 9216443 TI - Fibromuscular dysplasia of the renal artery and adrenal adenoma in a 36-year-old woman with hypertension. AB - A 36-year-old woman had fibrous dysplasia of the left renal artery and an aldosterone-producing adenoma of the right adrenal gland. The patient was evaluated first for secondary hypertension using a renal angiogram that showed fibrous dysplasia with stenosis of the left renal artery; angioplasty was successful. However, 1 month after angioplasty, hypertension recurred. Initially, it was thought that it had restenosed but after a negative angiography, adrenal computed tomography showed a 0.8-cm x 1-cm tumor of the right adrenal gland. The tumor was removed surgically, markedly improving her hypertension. PMID- 9216444 TI - Standardization of APTT reagents for heparin therapy monitoring: urgent or fading priority? PMID- 9216445 TI - Mass-spectrometric approaches for DNA-based genetic screening. PMID- 9216446 TI - Doping in sport: misuse, analytical tests, and legal aspects. PMID- 9216447 TI - Methods for detection of point mutations: performance and quality assessment. IFCC Scientific Division, Committee on Molecular Biology Techniques. AB - We give an overview of current methods for the detection of point mutations as well as small insertions and deletions in clinical diagnostics. For each method, the following characteristics are specified: (a) principle, (b) major modifications, (c) maximum fragment size that can be analyzed, (d) ratio and type of mutations that can be detected, (e) minimum ratio of mutant to wild-type alleles at which mutations can be detected, and (f) detection methods. Special attention is paid to the possibilities of quality assessment and the potential for standardization and automation. PMID- 9216448 TI - Screening blood spots for inborn errors of metabolism by electrospray tandem mass spectrometry with a microplate batch process and a computer algorithm for automated flagging of abnormal profiles. AB - Metabolic profiling of amino acids and acylcarnitines from blood spots by automated electrospray tandem mass spectrometry (ESI-MS/MS) is a powerful diagnostic tool for inborn errors of metabolism. New approaches to sample preparation and data interpretation have helped establish the methodology as a robust, high-throughput neonatal screening method. We introduce an efficient 96 well-microplate batch process for blood-spot sample preparation, with which we can obtain high-quality profiles from 500-1000 samples per day per instrument. A computer-assisted metabolic profiling algorithm automatically flags abnormal profiles. We selected diagnostic parameters for the algorithm by comparing profiles from patients with known metabolic disorders and those from normal newborns. Reference range and cutoff values for the diagnostic parameters were established by measuring either metabolite concentrations or peak ratios of certain metabolite pairs. Rigorous testing of the algorithm demonstrates its outstanding clinical sensitivity in flagging abnormal profiles and its high cumulative specificity. PMID- 9216449 TI - Simple triple-label detection of seven cystic fibrosis mutations by time-resolved fluorometry. AB - We describe a simple hybridization assay performed in microtitration wells with use of DNA probes labeled with three different lanthanide chelates for detection of seven mutations that cause cystic fibrosis. The assay is based on DNA amplification of four fragments containing the mutations (delta F508, G1717-->A, G542X, R553X, 3905 insertion T, W1282X, and N1303K) by PCR, followed by hybridization with short, allele-specific oligonucleotide probes labeled with europium, terbium, or samarium chelates. Because the technology makes it possible to hybridize three DNA probes simultaneously in one reaction, all 14 mutation related alleles were detected in a total of five reaction wells. Blood spot specimens, obtained from children with cystic fibrosis, their parents, and their siblings, have been assayed, and for all the probes the positive signal-to-noise ratios are > 10. Solution hybridization utilizing triple-label time-resolved fluorometry combined with PCR is a suitable procedure for large-scale screening and automation. PMID- 9216450 TI - Detecting CFTR gene mutations by using primer oligo base extension and mass spectrometry. AB - A new method for the reliable identification of localized variations in DNA by detection of associated diagnostic products with matrix-assisted laser desorption ionization time-of-flight mass spectrometry is described. The diagnostic products are generated by the primer oligo base extension (PROBE) reaction, which requires a single detection primer complementary to a region down-stream of a target strand's variable site. On addition of a polymerase, three dNTPs, and the fourth nucleotide in dideoxy form, the primer is extended through the mutation region until the first ddNTP is incorporated; the mass of the extension products determines the composition of the variable site. Tests for five cystic fibrosis mutations, including two exon 11 sites measured in a biplex reaction, and for differentiating between three common alleles of the poly(T) tract at the intron 8 splice acceptor site of the CFTR gene are presented. All experimental steps required for PROBE are amenable to the high degree of automation desirable for a high-through-put diagnostic setting. Furthermore, it requires no fluorescent, chemiluminescent, or radioactive labeling; the mass signals measured offer a far more analytically definitive signal, leading in all cases to high-quality unambiguous and easily interpreted results. PMID- 9216451 TI - Direct colorimetric monoclonal antibody enzyme immunoassay for estradiol-17 beta in saliva. AB - We developed a direct microtiter plate enzyme immunoassay to measure estradiol-17 beta in saliva. The assay has a commercially available monoclonal antibody, raised against estradiol-17 beta-6-carboxymethyloximebovine serum albumin, and a homologous horseradish peroxidase conjugate measured colorimetrically. The detection limit (equivalent to B0-3 SD) is 365 amol/well or 7.3 pmol/L when 50 microL samples are assayed. Cross-reactivity with estrone and estriol, testosterone, or progesterone is < 0.2%. Estradiol-17 beta was measured in daily samples over five natural menstrual cycles and eight cycles stimulated as a preliminary to in vitro fertilization, and the concentrations and fluctuations found agreed with previously published data. This method gives results in approximately 3 h and may be useful for fertility monitoring and management. PMID- 9216452 TI - Analytical and clinical evaluation of the Immulite estradiol assay in serum from patients undergoing in vitro fertilization: estradiol increase in mature follicles. AB - We examined an immunoassay for estradiol (E2) on the Immulite-an automated, random access chemiluminescent immunoassay system-to determine its accuracy and precision required for in vitro fertilization (IVF) studies. The assay, which has a reportable range from 73 to 7300 pmol/L, demonstrated good linearity under dilution, a detection limit of 44 pmol/L, and interassay CVs of 12.6% and 7.6% at 466 and 6164 pmol/L, respectively. In a retrospective analysis of 545 serum samples, the assay showed adequate agreement with an antibody-coated-tube RIA. The two E2 assays showed good agreement, even on samples from patients receiving a variety of different estrogen replacement therapies. Longitudinal studies of individual IVF cycles showed good parallelism between the automated system and the RIA, and results by the automated assay correlated well with the total number of follicles. PMID- 9216453 TI - Mass spectrometric characterization of nicked fragments of the beta-subunit of human chorionic gonadotropin. AB - We describe the use of an HPLC/MS technique for the characterization of nicked fragments of hCG beta-subunit. After reductive alkylation of the nicked hCG beta subunit with vinylpyridine, endoproteinase Glu-C or trypsin was used to digest the protein to produce peptides that could be analyzed by HPLC/electrospray ionization MS. Human leukocyte elastase digestion was used to produce an experimentally nicked hCG. Two nicking sites were observed, between amino acids 42Thr and 43Arg and between 44Val and 45Leu. The former site has not been previously reported for elastase digestion. The structures of the fragments were confirmed by HPLC/MS after removal of the oligosaccharide by direct mass measurement and by mass determination of their proteolytic digests. Without the glycopeptidase treatment, the microheterogeneity of the two N-linked oligosaccharides could be deduced from the spectra of the proteolytic fragments. Nicking with elastase was found to alter the oligosaccharide structures. Nicked beta-subunit samples isolated from the urine of choriocarcinoma patients were also analyzed and the location of the nicking site(s) agreed with that determined by classical techniques. Important differences in the oligosaccharide structures were also observed in these samples, including the presence of triantennary oligosaccharides not found in hCG from healthy subjects. These findings demonstrate the potential of HPLC/MS for characterization of glycoprotein standard preparations. PMID- 9216454 TI - Serum enzymes in heat stroke: prognostic implication. AB - We measured serum creatine kinase (CK), lactate dehydrogenase (LD), aspartate aminotransferase (AST), and serum alanine aminotransferase (ALT) in 26 heat stroke (HS) victims and 10 control (non-heat-exhausted) subjects during annual Hajj in Makkah, Saudi Arabia. On admission to the HS treatment unit, serum CK, AST, ALT, and LD were higher in HS victims than controls (P < 0.05), and at 6, 12, and 24 h were higher than baseline concentration. The patient group was divided into three groups, (a) those who had a quick recovery, (b) those who were critically ill until the end of the Hajj period (7 days), and (c) those who died. Serum enzymes at the time of admission were significantly higher (P < 0.05) in the nonsurviving group (n = 6) and the severely ill (n = 9) than in those who had a quick recovery (n = 11). ROC curves were plotted for each enzyme. The most useful indicator was LD, as it could distinguish significantly between the groups who died and those who had a quick recovery (area under the curve = 0.991 +/- 0.0286). It was followed by CK and AST as useful prognostic factors. When compared with ROC curves for body temperature, anion gap, and serum potassium, the enzyme results were superior prognostic indicators. PMID- 9216455 TI - Serum concentrations of free ubiquitin and multiubiquitin chains. AB - Ubiquitin, which can conjugate with cellular proteins, is classified into two forms: free ubiquitin and multiubiquitin chains. The latter is active as a signal for degradation of the targeted proteins. We found both forms in human serum and, using two immunoassays, quantitated them in sera from healthy subjects and patients with some diseases. Because of putative leakage of erythrocyte ubiquitin, hemolytic serum and serum obtained after long incubation (> 1-2 h) of blood at room temperature were excluded. Serum concentrations of multiubiquitin chains and free ubiquitin were substantially higher in rheumatoid arthritis and hemodialysis patients, respectively, than healthy subjects. Additionally, in acute viral hepatitis, serum multiubiquitin chain concentrations were increased in the acute phase, decreased in the recovery phase, and correlated with alanine and aspartate aminotransferase activities (r = 0.676 and 0.610, P < 0.0001 and < 0.001, respectively). Therefore, serum ubiquitin may have prognostic value. PMID- 9216456 TI - Correlation between plasma 5-aminolevulinic acid concentrations and indicators of oxidative stress in lead-exposed workers. AB - 5-Aminolevulinic acid (ALA), a heme precursor accumulated in acute intermittent porphyria and lead poisoning, undergoes metal-catalyzed aerobic oxidation at physiological pH to yield reactive free radical species (O2., HO., and ALA.). We analyzed the relationships between plasma ALA concentrations, blood concentrations of lead, protoporphyrin IX (PP-IX), superoxide dismutase (SOD), and methemoglobin (metHb), and urine chemiluminescence (CL) in samples collected from lead-exposed workers. All variables measured were substantially (P < 0.01) higher (2-8-fold) in the lead-exposed workers (n = 60). Plasma ALA concentrations were, on average, 6-fold higher in lead-exposed workers. We observed positive linear relationships between ALA and lead (r = 0.992), ALA and PP-IX (r = 0.891), ALA and metHb (r = 0.984), lead and SOD (r = 0.948), ALA and urine CL (r = 0.987), and lead and PP-IX (r = 0.993). These data are consistent with our free radical hypothesis for lead poisoning, where ALA distribution to and accumulation in several organs may trigger oxidative stress responses. PMID- 9216457 TI - Analytical performance of the Tandem-R free PSA immunoassay measuring free prostate-specific antigen. AB - The analytical performance of the Tandem-R free PSA assay available from Hybritech Inc. was evaluated. Comparison of recoveries of purified free (unbound) prostate-specific antigen (PSA) diluted in female serum in the Tandem-R free PSA assay and the Tandem-R (total) PSA assay demonstrated a link in calibration between the assays and an accurate determination of percent free PSA. The cross reactivity of the assay to purified PSA-alpha 1-antichymotrypsin was determined to be < 1%. The minimum-detectable concentration was < 0.05 microgram/L. The within-run and between-day CVs were < or = 5% for samples with > 0.3 microgram/L free PSA. Dilution and recovery showed no significant deviations from linearity across the assay range. The assay was insensitive to interference from blood components. The Tandem-R free PSA kit was shown to be an accurate, precise, and reliable assay for the measurement of free PSA. PMID- 9216458 TI - Plasma malondialdehyde as biomarker for oxidative stress: reference interval and effects of life-style factors. AB - Malondialdehyde (MDA) is one of the most frequently used indicators of lipid peroxidation. To generate reliable reference intervals for plasma malondialdehyde (P-MDA), a reference sample group was established in Funen, Denmark. The group consisted of 213 individuals (107 men, 106 women), ages 20-79 years. P-MDA was measured in EDTA-treated plasma after derivatization by thiobarbituric acid (TBA) and separation on HPLC. UV detection was performed at 532 nm. A reference interval was calculated as recommended by IFCC with REFVAL 3.42. The estimated reference limits (0.025 and 0.975 fractals) for the group were 0.36 and 1.24 mumol/L. The data were analyzed for gender- and age-related differences. Analysis of variance showed no interaction between gender and age, but separate analyses showed an independent effect of gender (P = 0.03), but not of age (P = 0.11). Daily smokers had a slightly higher average concentration of P-MDA than nonsmokers (P = 0.05), and P-MDA correlated with daily exposure to cigarette smoke (r = 0.162; P = 0.03). A positive correlation was also demonstrated between P-MDA and weekly alcohol consumption (r = 0.153; P = 0.03). Within-subject and day-to-day variations of P-MDA indicated that the potential of P-MDA as a biomarker for individuals is questionable. However, on a group basis, the present data support that P-MDA may be a potential biomarker for oxidative stress. PMID- 9216459 TI - Preparation of lyophilized partial thromboplastin time reagent composed of synthetic phospholipids: usefulness for monitoring heparin therapy. AB - To contribute to the development of a reference reagent for monitoring heparin therapy, a lyophilized partial thromboplastin time (PTT) reagent was prepared from synthetic dioleoylphosphatidylcholine, dioleoylphosphatidylserine, and dioleoylphosphatidylethanolamine, with colloidal silica as activator. The reagent, coded 91/558, was contained in sealed glass ampoules; it deteriorated in a heat degradation experiment, but its activity remained constant for at least 4 years when stored at -70 degrees C. Within- and between-run precision with this reagent complied with the requirements proposed by the International Committee for Standardization in Haematology (ICSH) Panel on PTT. The response of this reagent and of two other reagents to heparin added to pooled normal plasma was nonlinear. Citrated samples from 58 patients receiving intravenous heparin and from 24 apparently healthy volunteers were tested with reagent 91/558, with Automated APTT (Organon Teknika), with Manchester APTT reagent, with an antifactor Xa assay, and with an anti-factor IIa assay. The correlation of APTT with anti-Xa and anti-IIa activity was poor. The best correlation was observed between reagent 91/558 and the Organon Teknika reagent. Correlations were improved when individual patients' samples were replaced by pooled plasmas from heparinized patients, in whom the effect of oral anticoagulation was minimal. These results suggest that preparation of a lyophilized synthetic phospholipid reagent is feasible for use in monitoring heparin therapy. PMID- 9216460 TI - Plasma protein abnormalities in nephrotic syndrome: effect on plasma colloid osmotic pressure and viscosity. AB - The concentrations of 25 plasma proteins were measured in 22 patients with membranous nephropathy. For some large proteins, the plasma concentrations were increased; there were also large proteins with low plasma concentrations, but small or medium-sized proteins showed uniformly lower plasma concentration than the controls. Plasma colloid osmotic pressure (pi) and viscosity (eta) were not interrelated but showed positive and significant correlations with plasma concentrations of small and medium-sized proteins (pi) and plasma concentrations of large proteins (eta), respectively. Nephrotic plasma is not efficient in maintaining plasma pi but highly efficient in maintaining plasma eta. High plasma fibrinogen concentrations and low antithrombin III concentrations may predispose to thrombosis, and low IgG concentrations may account for the higher predisposition to bacterial infection. The relative composition of nephrotic plasma is heavily dependent on the size of the different proteins. Plasma pi and eta are also maintained by the relative preponderance of different plasma proteins. PMID- 9216461 TI - Some historical perspectives on AACC. PMID- 9216462 TI - Comparison of commercial screening tests for glucose-6-phosphate dehydrogenase deficiency in the neonatal period. PMID- 9216463 TI - Topical application of eosin to burns produces interference in measurement of serum vancomycin by fluorescence polarization immunoassay. PMID- 9216464 TI - Five osteocalcin assays compared: tracer specificity, fragment interference, and calibration. PMID- 9216465 TI - Alternative, noninvasive tissues for quantitative screening of mutant mitochondrial DNA. PMID- 9216466 TI - Plasma renin activity: temperature optimum at approximately 45 degrees C. PMID- 9216467 TI - Urine screening for flunitrazepam: applicability of Emit immunoassay. PMID- 9216468 TI - Hydrogen peroxide interferes with detection of nitric oxide by an electrochemical method. PMID- 9216470 TI - A nonoccupational source of mercury intoxication. PMID- 9216469 TI - Blood lead screening (continued). PMID- 9216471 TI - Salicylate interference with the Roche Cobas Integra chloride electrode. PMID- 9216473 TI - Decline in blood lead in Ontario children correlated to decreasing consumption of leaded gasoline, 1983-1992. PMID- 9216472 TI - Falsely increased serum estradiol results reported in direct estradiol assays. PMID- 9216475 TI - Issues in detecting abuse of xenobiotic anabolic steroids and testosterone by analysis of athletes' urine. AB - Over the last decade the number of laboratories accredited by the International Olympic Committee (IOC) has grown to 25. Nearly half of the approximately 90,000 samples tested annually are collected on short notice-the most effective means to deter the use of anabolic androgenic steroids (AAS). The major urinary metabolites of AAS have been characterized and are identified by their chromatographic retention times and full or partial mass spectra. The process of determining if an athlete has used testosterone (T) begins with finding a T to epitestosterone (E) ratio > 6 and continues with a review of the T/E-time profile. For the user who discontinues taking T, the T/E reverts to baseline (typically approximately 1.0). For the extremely rare athlete with a naturally increased T/E ratio, the T/E remains chronically increased. Short-acting formulations of T transiently increase T/E, and E administration lowers it. Among ancillary tests to help discriminate between naturally increased T/E values and those reflecting T use, the most promising is determination of the carbon isotope ratio. PMID- 9216474 TI - Hormonal doping and androgenization of athletes: a secret program of the German Democratic Republic government. AB - Several classified documents saved after the collapse of the German Democratic Republic (GDR) in 1990 describe the promotion by the government of the use of drugs, notably androgenic steroids, in high-performance sports (doping). Top secret doctoral theses, scientific reports, progress reports of grants, proceedings from symposia of experts, and reports of physicians and scientists who served as unofficial collaborators for the Ministry for State Security ("Stasi") reveal that from 1966 on, hundreds of physicians and scientists, including top-ranking professors, performed doping research and administered prescription drugs as well as unapproved experimental drug preparations. Several thousand athletes were treated with androgens every year, including minors of each sex. Special emphasis was placed on administering androgens to women and adolescent girls because this practice proved to be particularly effective for sports performance. Damaging side effects were recorded, some of which required surgical or medical intervention. In addition, several prominent scientists and sports physicians of the GDR contributed to the development of methods of drug administration that would evade detection by international doping controls. PMID- 9216476 TI - Endocrine aspects of anabolic steroids. AB - Understanding of the mechanism of androgen action has been enhanced by advances in knowledge on the molecular basis of activation of the androgen receptor and the importance of tissue conversion of circulating testosterone to dihydrotestosterone and estradiol. New evidence supports the view that supraphysiological doses of anabolic steroids do have a definite, positive effect on muscle size and muscle strength. However, the nature of the anabolic action of androgens on muscle is currently unclear and may involve mechanisms independent of the androgen receptor. The dose-response relationships of anabolic actions vs the potentially serious risk to health of androgenic-anabolic steroids (AAS) use are still unresolved. Most of the adverse effects of AAS are reversible but some are permanent, particularly in women and children. The reported incidence of acute life-threatening events associated with AAS abuse is low, but the actual risk may be underrecognized or underreported; the exact incidence is unknown. The long-term consequences and disease risks of AAS to the sports competitor remain to be properly evaluated. PMID- 9216477 TI - Immunoprocedures for detecting human chorionic gonadotropin: clinical aspects and doping control. AB - The pregnancy hormone human chorionic gonadotropin (hCG) is also present at low concentrations in plasma and urine of men and nonpregnant women. hCG immunoreactivity occurs in various molecular forms: Besides the intact hCG heterodimer, considerable amounts of proteolytically cleaved forms, free subunits, and fragments are found in plasma and urine. Especially in urine, proteolytic fragments constitute a major part of the hCG immunoreactivity. The different forms of hCG cross-react to various degrees in immunoassays and constitute a problem for standardization of specific hCG determinations. After injection of hCG (10,000 IU of Pregnyl; Organon), above-normal concentrations of hCG can be detected in serum and urine for 7-11 days. Most immunoassays for hCG also measure hCG beta. Quantitative hCG determinations are mainly performed on serum samples, and very few commercial hCG determinations have been validated for determination of urine samples. Considerable care must therefore be exercised when utilizing such assays to analyze urines for doping control. PMID- 9216478 TI - Analytical advances in detection of performance-enhancing compounds. AB - The use and abuse of performance-enhancing substances has been an issue in sports since the ancient Greeks. The availability of numerous synthetic steroids and recombinant peptide hormones has made testing an analytical challenge. Recent advances in mass spectrometry have provided an opportunity to decrease detection limits. The Atlanta Olympic Games in 1996 marked the first time every specimen was screened by gas chromatography (GC) coupled to high-resolution mass spectrometry (MS). A further improvement may be seen with GC/MS/MS and quadrupole ion traps. Electrospray HPLC/MS has also been applied to the detection and confirmation of peptide hormones in urine. The ability to detect subtle differences in oligosaccharide structure may provide a way to detect abuse of recombinant glycoproteins. Simply decreasing detection limits is not enough; new technology also allows development of a foundation on which to base interpretation. Application of HPLC/MS/MS has allowed direct measurement of steroid conjugates in urine. The relative importance of sulfate, glucuronide, and other conjugates and metabolites of testosterone and epitestosterone can now be assessed. In the international sports arena, the impact of genetic metabolic disposition must also be considered if we are to provide an equitable system. Further research will establish more-refined criteria for the detection threshold of abused substances. PMID- 9216479 TI - Electrocortical and behavioral responses produced by acute electrical stimulation of the human centromedian thalamic nucleus. AB - Incremental, desynchronizing and spike-wave electrocortical responses and concomitant symptoms to acute electrical stimulation of the centromedian thalamic nucleus (CM) were studied in 12 patients with intractable complex partial and tonic-clonic generalized seizures. Low-frequency (6/s), 320-800 microA stimulation of the caudal-basal and central portions of CM elicited incremental recruiting and augmenting-like responses with a bilateral regional scalp distribution, with emphasis at the ipsilateral frontal (recruiting) and central (augmenting) regions, while ventral-basal CM stimulation elicited primary-like responses with a focal distribution at the ipsilateral parietal region. High frequency (60/s), 320-800 microA stimulation of caudal-basal and central, but not ventral-basal CM, elicited EEG desynchronization and a slow negative shift of the EEG baseline with scalp distribution similar to that showed by recruiting- and augmenting-like responses. Neither incremental nor desynchronization EEG responses were accompanied by evident patient sensory or motor responses. Low frequency (3/s), high-intensity (30 V = 2400 microA) combined stimulation of the right CM and left non-specific mesencephalic ascending pathways elicited a response similar to the typical absence attack with all EEG and clinical ingredients: S1, S2, P1 and W components of the individual spike-wave complex and generalized spike-wave discharges followed by sleep spindle EEG afterdischarges, accompanied by motionless stare, 3/s eye blinking, lip smacking and total failure to respond to visual stimuli in patients under a simple responding task. PMID- 9216480 TI - A method to quantify invariant information in depth-recorded epileptic seizures. AB - In the field of epilepsy, the analysis of stereoelectroencephalographic (SEEG) signals recorded with depth electrodes provides major information on interactions between brain structures during seizures. A methodology of comparing SEEG seizure recordings is applied in 4 patients suffering from temporal lobe epilepsy. It proceeds in 3 steps: (i) segmentation of SEEG signals, (ii) characterization and labeling of segments and (iii) comparison of observations coded as sequences of symbol vectors. The third step is based on a vectorial extension of Wagner and Fischer's algorithm to first, quantify similarities between observations and second, extract invariant information, referred to as spatio-temporal signatures. These are automatically extracted by the algorithm without the need to make a priori assumptions on the 'patterns' to be searched for. Theoretical results show that two observations of non-equal duration can be matched by deforming the first one (using insertion/deletion operations on vectors) to optimally fit the second, under a minimal cost constraint. Clinical results show that the study brings objective results on reproducible mechanisms occurring during seizures: for a given patient, quantified descriptions of seizure periods are compared and similar ictal patterns, or signatures, are extracted from SEEG signals. Some of these signatures (particularly those containing spikes, spike-and-waves, slow waves and rapid discharges) are relevant: they seem to reflect reproducible propagation schemes whose analysis may help in the understanding of epileptogenic networks. PMID- 9216481 TI - Mesial temporal versus lateral temporal interictal epileptiform activity: comparison of chronic and acute intracranial recordings. AB - Intracranial interictal epileptiform activity (EA) was recorded by chronic stereotactic depth electroencephalography (SDEEG) and acute electrocorticography (ECOG) in 22 patients with complex partial seizures of temporal lobe origin. Chronic SDEEG recordings defined two groups of patients with respect to the presence or absence of lateral temporal EA; 13 patients showed independent lateral temporal EA during chronic recordings and 9 patients did not. All patients had EA recorded from mesial temporal structures during SDEEG. The presence of lateral temporal EA was correlated with a higher pre-operative seizure frequency but not with ictal onset zones, structural pathology, age at onset of epilepsy, or duration of epilepsy. Results of acute ECOG recordings performed on the same patients 1-24 months after SDEEG accurately reproduced the mesial versus lateral distribution of EA within patients (P < 0.0003). Though ECOG was less sensitive than SDEEG in demonstrating EA confined to mesial structures, positive findings at ECOG were 100% specific with respect to SDEEG. These results suggest that, at least with respect to mesial temporal versus lateral temporal structures, there is a constancy within patients in the distribution of interictal EA recorded with chronic intracranial electrodes. In addition, acute ECOG provides an accurate representation of individual patients' interictal EA. PMID- 9216482 TI - The effects of external load on movement-related changes of the sensorimotor EEG rhythms. AB - The effects of external load opposing brisk voluntary extension of the right index finger on the EEG rhythms in the left and right sensorimotor hand area were studied in 13 right-handed subjects. Four levels of external loads corresponding to the weights of 0 g (no load), 30 g, 80 g and 130 g were used. The effects of external load on EEG rhythms were the following: (i) prior to movement, the desynchronisation of beta-rhythms (18-25 Hz) over the contralateral sensorimotor area was greater under the two largest loads as compared to the 0 g load. However, beta-desynchronisation at ipsilateral electrodes was larger under the 80 g load than under the 130 g load, presumably due to a transcallosally mediated inhibition exerted by the highly excited contralateral motor area; (ii) the mu rhythm desynchronisation continued over both hemispheres for about 0.3-0.4 s after movement and the largest load was accompanied by the longest mu-rhythm desynchronisation; (iii) the post-movement beta-synchronisation was also longer under the heaviest load (130 g) as compared to the no-load condition (0 g), especially in subjects who prolonged their total movement time under the heaviest load. The results show that (i) the movement-related desynchronisation and synchronisation of sensorimotor EEG rhythms is influenced by external load opposing finger movement, and (ii) the effects of external load differ for the mu and beta-rhythms. PMID- 9216483 TI - Motor cortical reflex myoclonus: a case study with MEG. AB - Cortical reflex myoclonus usually depends for its generation on the hyperexcitability of sensory cortex, which manifests itself as an enhanced somatosensory evoked potential (SEP). A 25-year-old female, presenting with involuntary jerky dorsiflexion of the left foot, was found to have cortical reflex myoclonus which was aggravated during intended movements. The jerks were also elicited by electrical stimulation of the posterior tibial nerve, although the SEP evoked by the same stimulus was normal in latency and amplitude. Both the spontaneous spikes and the premyoclonus spike demonstrated by jerk-locked back averaging were localized to the superior frontal gyrus, just anterior to the paracentral sulcus. Paired-pulse magnetic stimulation disclosed lack of inhibition in the right hemisphere leg motor area, whereas the excitability of sensory cortex as studied by paired SEP testing was normal. This suggests that the myoclonus was caused mainly by enhanced excitability of the motor cortex and that this alone was enough for the production of long loop reflexes. We propose to designate this type of cortical myoclonus as motor cortical reflex myoclonus. It is generated in the motor cortex, but is at the same time stimulus-sensitive. PMID- 9216484 TI - Poverty, cultural disadvantage and brain development: a study of pre-school children in Mexico. AB - Forty-two children, who had been studied previously at the age of 18-30 months, were studied again at 4 years of age. Twenty-two belonged to low socioeconomic strata and were classified as high-risk children (HRC) the other 20 were classified as low-risk children (LRC), and belonged to middle and middle-high socioeconomic strata. Ten minutes of EEG using reference derivations (with linked earlobes) were recorded from each subject. Twenty EEG segments of 3.2 s each were selected by visual inspection for Fourier analysis. Absolute power (AP) was computed for the total EEG energy (1.5-19 Hz) as well as each reference derivation in 4 frequency bands: delta (1.5-3.5 Hz), theta (4-7.5 Hz), alpha (7.5 12.5 Hz) and beta (12.5-19 Hz). HRC had significantly more delta AP than LRC in frontal and central leads, and higher values of theta AP in frontal leads. Alpha AP was higher in LRC in occipital areas and in F8 and T4. This study suggests a maturational lag in HRC. PMID- 9216485 TI - Functional involvement of central nervous system in mitochondrial disorders. AB - Thirty-nine patients with mitochondrial diseases were studied with somatosensory and motor evoked potentials. Sixteen patients (41%) had clinical and 12 (31%) had neuroradiological evidence of central nervous system involvement. The overall incidence of electrophysiological abnormalities was 64%. Abnormal evoked potentials were also found in a significant percentage (33%) of patients with pure myopathic forms of mitochondrial diseases and in an asymptomatic carrier of MERRF mutation. Of the individual tests, somatosensory evoked potentials were abnormal in 49% of the patients and motor evoked potentials were abnormal in 46% of the patients. The outcome is that electrophysiological evidence of central nervous system involvement is present in a high percentage of patients with mitochondrial disorders, and that the threshold for central nervous system electrophysiological abnormalities is well below that for clinical and/or radiological manifestations. PMID- 9216486 TI - Motor unit size estimation: confrontation of surface EMG with macro EMG. AB - Surface EMG (SEMG) is little used for diagnostic purposes in clinical neurophysiology, mainly because it provides little direct information on individual motor units (MUs). One of the techniques to estimate the MU size is intra-muscular Macro EMG. The present study compares SEMG with Macro EMG. Fifty eight channel SEMG was recorded simultaneously with Macro EMG. Individual MUPs were obtained by single fiber triggered averaging. All recordings were made from the biceps brachii of healthy subjects during voluntary contraction at low force. High positive correlations were found between all Macro and Surface motor unit potential (MUP) parameters: area, peak-to-peak amplitude, negative peak amplitude and positive peak amplitude. The MUPs recorded with SEMG were dependent on the distance between the MU and the skin surface. Normalizing the SEMG parameters for MU location did not improve the correlation coefficient between the parameters of both techniques. The two measurement techniques had almost the same relative range in MUP parameters in any individual subject compared to the others, especially after normalizing the surface MUP parameters for MU location. MUPs recorded with this type of SEMG provide useful information about the MU size. PMID- 9216487 TI - Clinical value of F-wave recordings in traumatic cervical spinal cord injury. AB - F-waves and motor/sensory nerve conduction (NCS) of the median and ulnar nerves were examined in 66 patients with traumatic motoneurone lesion due to acute and chronic cervical spinal cord injury (SCI). The examinations were performed in parallel in chronic tetraplegics once and in acute tetraplegic patients monthly for the first 3 months, after 6 months and 1 year post-trauma. A pathological reduction of the compound muscle action potential (CMAP) (in 10% even a complete loss of the CMAP) was present in about 50% of the patients. The mean CMAP values of tetraplegic patients with either acute or chronic SCI were significantly (P < 0.001) reduced compared to normal subjects. Because sensory nerve conduction in these patients was normal, the reduction of CMAP should be due to damage of intramedullar motoneurones or anterior nerve roots. While in all chronic SCI patients with preserved CMAP F-waves could be elicited, 50% of the acute SCI patients showed a complete loss of F-waves of both nerves during the initial examination due to spinal shock. After 6 months all acute SCI patients with preserved motor potentials regained F-waves. Therefore, the excitability of F waves is influenced by spinal shock in acute SCI. The mean F-wave latencies (Fmin response, Fmin-M response) revealed no significant difference between healthy subjects and SCI patients. However, the frequency of F-wave production was related to the severity of the motoneurone lesion. Furthermore, while the F-wave latencies and CMAP values did not change significantly with time after acute SCI, the frequency of F-wave production increased, but remained reduced compared to normal subjects. PMID- 9216488 TI - Reflexes evoked in human erector spinae muscles by tapping during voluntary activity. AB - We studied the stretch reflexes, an early R1 and a late R2, by tapping the voluntarily contracted erector spinae muscles and recording from the same spinal level with the subject in the prone position. The onset latencies increased progressively towards the caudal level from 8.8 +/- 0.7 ms at T5-6 to 15.9 +/- 1.1 ms at L4-5 for R1, and from 33.3 +/- 2.7 ms at T5-6 to 49.1 +/- 2.8 ms at L4 5 for R2. The latency changed significantly (P < 0.05) between two adjacent segments from T5-6/T6-7 through L1-2/L2-3 for R1 and T5-6/T6-7 through L3-4/L4-5 for R2. When recorded remote from the stimulus site, R1, considered segmental in origin, showed, as expected, only a small latency change consistent with the time required for the mechanical event to propagate to the recording site. In contrast R2 was shorter in latency with more rostral stimulation irrespective of the distance to the recording sites. This finding implies a centripetal propagation of the afferent impulse along the central pathway, which shortens with more rostral site of stimulation. Of the two components, the more reproducible R1 has a potential diagnostic value for segmental evaluation of thoracic nerve root compression and truncal neuropathies. PMID- 9216489 TI - Sequential changes of auditory processing during target detection: motor responding versus mental counting. AB - Brain potentials evoked to non-targets in an auditory target detection task changed in amplitude, duration, polarity, and scalp topography as a function of position in the stimulus sequence relative to the target. (1) A negative prestimulus readiness like-potential, or RP, the poststimulus N100, and a late slow wave to non-targets immediately after the target were reduced in amplitude compared to non-targets immediately before the target. The amplitudes of these potentials after the target then increased in size as a linear function of the number of non-targets in the sequence. (2) The amplitudes of the positive components, P50 and P200, were larger to non-targets immediately after the target than to non-targets immediately before the targets. P50 amplitude then decreased to subsequent non-targets in the sequence in a linear manner; P200 amplitude was reduced equivalently to all subsequent non-targets. (3) The duration of the P200 component could extend into the time domain when the P300 to targets would occur. The P200 component to non-targets was therefore designated 'P200/300'. The duration of the P200/300 component was shorter to non-targets immediately after the target than to non-targets immediately before the targets. P200/300 duration then extended in a linear manner to subsequent non-targets in the sequence and approached the peak latency of the P300 evoked by targets. (4) The anterior/posterior scalp distribution of P50 and the polarity of the late slow wave to non-targets changed as a function of non-target position in the sequence. The subject's response to the targets (button press or mental count) influenced these sequential effects. Linear trends for sequence were present in the press but not the count conditions for the amplitude of the RP, N100, and P300; linear trends for P50, P200/300 duration, and the late slow wave were found in both the press and count conditions. Reaction time was speeded as a function of the number of preceding targets. These dynamic changes in the processing of auditory signals were attributed to an interaction of attention and the subjective expectancies for both the appearance of a target stimulus and the requirement to make a motor response. PMID- 9216490 TI - Clinical multisegmental posturography: age-related changes in stance control. AB - Unlike conventional platform posturography, which analyses the sway in the projection of the body baricentre on a supporting plane, multisegmental posturography provides information about body segmental movements during stance, including those that keep the baricentre still. This paper presents a new technical approach to multisegmental posturography using Virtual Reality electromagnetic tracking devices. This device was used to study age-related differences in normal subjects in the control of upright posture. Body sway was studied by recording the oscillations of two trackers placed on the head and the hip during the Romberg test. The tracking device allowed us to detect age-related differences in postural stance strategies. Although the amplitude and velocity of the oscillations measured at the head did not differ in the two groups, the flexibility of the ankle-hip head axis differed significantly: elderly subjects exhibited a more rigid stance. Closing the eyes increased rigidity in both age groups and this change appear more pronounced in the young. PMID- 9216491 TI - Task-dependence of muscle afferent monosynaptic inputs to human extensor carpi radialis motoneurones. AB - The task-dependence of homonymous muscle afferent inputs was investigated in motor units of the extensor carpi radialis muscles during voluntary isometric contraction involving either the activation of agonist extensor muscles (wrist extension) or the co-activation of antagonist extensor and flexor muscles (hand clenching). The effectiveness of the muscle afferent monosynaptic inputs was tested by delivering either tendon taps or electrical stimulation to the radial nerve. In both cases, the motor unit responses, which took the form of narrow peaks in the peri-stimulus time histograms, were found to be significantly greater during hand clenching. The parallel enhancement of the responses to both mechanical and electrical stimulations observed during hand clenching could not be explained in terms of changes in the muscle spindle responsiveness. The enhancement of the motor units' responsiveness was apparent during the first 0.5 ms of the peaks in the peri-stimulus time histograms, taken to be uncontaminated by any polysynaptic components. It may therefore have reflected an increase in the amplitude of the excitatory monosynaptic potentials generated by the muscle spindle primary afferents. This is interpreted in terms of changes in the presynaptic inhibition, which might be depressed as the result of the large-scale activation of palm and finger cutaneous afferents liable to occur during hand clenching. PMID- 9216492 TI - Silent period following transcranial magnetic stimulation: a study of intra- and inter-examiner reliability. AB - Transcranial magnetic stimulation-evoked silent period (SP) has been attributed largely to the activity of intracortical inhibitory systems and recent reports provided evidence that it is a useful indicator of central motor disturbances. We studied the intra- and inter-examiner reliability of SP measurements in 28 healthy subjects. In 15 subjects SP measurements were performed by one single examiner and repeated by the same examiner 3 days and 7 days later, showing a high degree of intra-examiner reliability over time. In another subgroup consisting of 13 volunteers SP measurements were performed on the same subject by three different examiners, demonstrating a higher level of variability. In both subgroups we found a high interindividual variability ranging from 44-258 ms and a considerably lower side-to-side difference within subjects. Our results suggest that longitudinal assessments of the SP in patients with central motor involvement should optimally be performed by a single examiner. Regarding the wide range of possible SP durations in healthy subjects the intraindividual side to-side symmetry seems to be the most valuable parameter. PMID- 9216493 TI - Absence of facilitation or depression of motor evoked potentials after contralateral homologous muscle activation. AB - We have previously described post-exercise facilitation and post-exercise depression of motor evoked potentials (MEPs) to transcranial magnetic stimulation (TMS). To determine the presence of post-exercise facilitation after exercise of a contralateral muscle, MEPs were recorded from the resting right extensor carpi radialis (ECR) muscle while the left ECR muscle was activated, then immediately after brief left ECR activation, and, finally, immediately after brief right ECR activation. We repeated the experiment using the first dorsal interosseous (FDI) muscle. To determine the presence of post-exercise depression after exercise of a contralateral muscle, MEPs were recorded from the right ECR after prolonged exercise of the left ECR, followed by right ECR recording after its fatigue. The mean MEP amplitudes from the right ECR and the right FDI after brief activation were 187% and 266% of their pre-exercise values, respectively. There were no significant changes in MEPs recorded from the right ECR or FDI muscles during or immediately after brief activation of their left counterparts. The mean amplitude of MEPs recorded from the right ECR after it fatigued was approximately half the pre-exercise value, but there was no significant change in MEPs recorded from the right ECR after prolonged exercise of the left ECR. Therefore, neither post exercise facilitation nor post-exercise depression occurred after contralateral homologous muscle exercise. PMID- 9216494 TI - Rapid-rate transcranial magnetic stimulation of human frontal cortex can evoke saccades under facilitating conditions. AB - Rapid-rate transcranial magnetic stimulation (rTMS) of a localized area within premotor frontal cortex elicited short-latency, multistep eye movements during a double-step saccade task in 3 of 9 subjects. These evoked saccades occurred in 14 32% of trials when rTMS was delivered over premotor regions located 5-7 cm lateral and 2-4 cm anterior to the vertex. In trials without rTMS or when rTMS was delivered medial or posterior to these sites, multistep saccades occurred in less than 2% of trials. In two subjects, rTMS of the right premotor cortex evoked contraversive saccades. In a third subject, rTMS of the left premotor cortex evoked saccades whose trajectory depended on the direction of the double-step targets. The amplitude and likelihood of occurrence of evoked saccades varied between rTMS trials. The intervals between evoked saccades were proportional to the intervals between rTMS pulses delivered at 16, 20 or 25 Hz. rTMS did not elicit saccades during fixation on a stationary target or during straight-ahead gaze in darkness, suggesting that the double-step saccade task facilitated the activation of a discrete premotor area consistent with the human frontal eye fields. PMID- 9216495 TI - Putting more prevention into your practice. PMID- 9216496 TI - Accuracy of frozen sections in assessing margins in oral cancer resection. AB - PURPOSE: This study examined the accuracy of frozen section diagnosis of tissue samples from surgical margins compared with the final histologic diagnosis of the same tissue. The total resection specimen was also examined to see whether frozen sections were helpful in predicting negative margins for the entire cancer. The nature of positive and negative margins and their implications for the surgeon are discussed. PATIENTS AND METHODS: The records of 49 consecutive patients with previously untreated squamous carcinoma of the mouth were reviewed. All frozen and permanent sections were evaluated by one pathologist. Margins involved by carcinoma, carcinoma in situ, dysplasia, or with carcinoma within 5 mm were defined as positive. Histologic findings were compared with the patient's clinical course to define the relationship between positive margins and local recurrence. Patients were followed for 17 to 45 months or until death. RESULTS: Three hundred four of 307 frozen sections showed concordance with the permanent section of the same tissue sample (two false negative and one false positive), an accuracy rate of 99%. When the final margins of the resected surgical specimen were compared with the frozen section diagnoses, ten patients had positive final margins. In three patients, these were diagnosed by frozen section. Seven patients had final margins that were positive when the surgical resection specimen was examined but were not diagnosed by frozen section. A greater local recurrence note was found in patients with invasive carcinoma at the margin, dysplastic margins, and margins within 5 mm of the cancer. CONCLUSIONS: Although frozen sections are extremely accurate, they are not as reliable in eliminating positive margins in the final specimen as the surgeon might hope. PMID- 9216497 TI - Positional changes in the mandibular condyle and amount of mouth opening after sagittal split ramus osteotomy with rigid or nonrigid osteosynthesis. AB - PURPOSE: The purpose of this study was to investigate postoperative positional changes in the mandibular condyle and mouth opening in patients undergoing sagittal split ramus osteotomy with either rigid or nonrigid osteosynthesis. PATIENTS AND METHODS: The forty-six patients with mandibular prognathism underwent sagittal split ramus osteotomy for mandibular set back followed by fixation with one of four methods: circumferential wire (n = 11), lag screw technique (n = 10), positional screw technique (n = 10), or miniplates (n = 15). The changes in the condylar position were assessed by measuring the angle of the condylar long axis (the condylar angle) on submentovertex radiographs. Mouth opening was evaluated by measuring the interincisal distance immediately after the release of maxillomandibular fixation and by monitoring the duration of trismus. RESULTS: Regardless of the procedure used the condylar angle increased in most patients after surgery (80 of 92 condyles). Although the amount of increase tended to be higher with rigid osteosynthesis than with nonrigid osteosynthesis, no significant differences were observed among the groups. Mouth opening was not significantly influenced by the type of osteosynthesis, and no patient complained of limitation 1 year after surgery. CONCLUSIONS: Although inward rotation of the condyle frequently occurs after osteosynthesis regardless of the procedure used, the changes in condylar position are within the range of adaptability of the patient. PMID- 9216498 TI - Calcifying odontogenic cyst: a clinicopathologic study of 57 cases with immunohistochemical evaluation for cytokeratin. AB - PURPOSE: A clinicopathologic study of all cases accessioned as calcifying odontogenic cyst (COC) from 1971 to 1996 from the files of the Oral Pathology Laboratory at Temple University School of Medicine was undertaken. MATERIALS AND METHODS: Microscopic slides and clinical histories of cases diagnosed as calcifying odontogenic cyst were reviewed and analyzed. Ten cases were processed for cytokeratin immunohistochemical staining. RESULTS: Fifty-seven cases were reviewed, 28 males and 29 females. Patients' ages ranged from 7 to 83 years, with a mean age of 49.8 years. Thirty-four cases involved the mandible and 23 were from the maxilla. Seventeen were reported in peripheral locations, and 38 occurred centrally within the jaws. Two were found both centrally and peripherally. The most common clinical sign for central lesions was a radiolucency sometimes associated with a jaw expansion. The most common clinical complaint for peripheral lesions was a nodular growth on the gingiva. Although lining epithelial cells were strongly positive for cytokeratin, full-brown ghost cells and disintegrating ghost cells were nonreactive. CONCLUSION: Calcifying odontogenic cyst can occur in any age-group, intraosseously or extraosseously, and as a solid lesion. No recurrences were found after surgical removal in the current series. PMID- 9216499 TI - Morbidity and mortality with outpatient anesthesia: the experience of a residency training program. AB - PURPOSE: Previous studies regarding anesthetic-related morbidity and mortality rates in the oral surgery office have usually taken the form of a survey. This retrospective investigation of outpatient anesthetic morbidity and mortality was undertaken to compare the safety record of an oral and maxillofacial surgery training program with that of private practitioners. MATERIALS AND METHODS: Records from all outpatient general anesthesia cases performed in the Department of Oral and Maxillofacial Surgery at the Boston University Goldman School of Graduate Dentistry between August 13, 1990, and September 30, 1994, were reviewed for the incidence of nineteen separate categories of morbidity. RESULTS: There were 1,126 general anesthetics performed. There were 26 recorded incidents of morbidity (2.3%), none of which resulted in any postoperative sequelae. There were no deaths. The most common complication encountered was laryngospasm, with nine recorded incidents (0.8%). The second most common complication was cardiac dysrhythmia with eight recorded incidents (0.8%). CONCLUSIONS: The low incidence of anesthetic-related morbidity seen in this study can most likely be attributed to proper patient selection. A carefully reviewed medical history and physical examination are the two most useful methods to prevent anesthetic emergencies. Another factor considered when selecting the proper anesthetic method includes the length and difficulty of the surgical procedure, with outpatient general anesthesia being reserved for those procedures that are predicted to be relatively short (30 to 45 minutes), and with little potential for airway difficulties. PMID- 9216500 TI - Cross-sectional area of the mandible. AB - PURPOSE: The anatomy of the mandible was examined by measuring the cross sectional area (CSA) of multiple regions of 10 fully dentulous hemimandibles to provide a better understanding of regional structural differences that may have implications regarding biomechanical strength, surgical reconstruction, and fracture site frequency. MATERIALS AND METHODS: Fifteen cuts from the condyle to the symphysis were made of each hemimandible (n = 150 cuts). A Zeiss Videoplan digitizer was used to determine the CSA. RESULTS: The total CSA through the condyle was greater than the CSA through the condylar neck. The CSA through the ramus exceeded that of the condylar neck. The total CSA of the midramus was significantly greater than that of the upper ramus. The total CSA at the body, parasymphysis, and symphysis was significantly greater than at the mid-angle. The total CSA of the cortex increased anteriorly; these differences become significant between the condylar neck and the body, parasymphysis, and symphysis. The total CSA, and the CSA of the cortex and spongiosa, remained relatively constant from the inferior angle anteriorly. CONCLUSIONS: Significant differences exist in the CSA at different points, with an increase in the total, cortical, and spongiosal CSA anteriorly from the condylar neck to the angle. The total CSA and the CSA of the cortex and spongiosa remain relatively constant anterior to the inferior angle. These data suggest that bony CSA alone is not the sole factor in determining fracture site frequency. PMID- 9216501 TI - Facial growth and the need for orthognathic surgery after cleft palate repair: literature review and report of 28 cases. AB - PURPOSE: Controversy still exists regarding the optimal timing and surgical technique for primary cleft lip and palate (CLP) repair, and treatment protocols vary considerably. This study reviews the literature on timing and technique for primary repair and reports on the outcome for a consecutive group of patients treated by a single surgical protocol at the Sunnyview Cleft Palate Clinic. PATIENTS AND METHODS: Twenty-eight patients treated by a standardized clinical protocol from infancy through adolescence were evaluated with respect to the need for orthognathic surgery to correct jaw size discrepancy. For each patient, data was collected regarding type of cleft deformity, total number of surgical procedures from infancy, surgeon performing the primary repair, and the need or indication for orthognathic surgery. RESULTS: Twenty-five percent of patients treated by this protocol required orthognathic surgery because of anteroposterior jaw size discrepancy. The number of prior operations was not a significant factor. The need for orthognathic surgery was seen in all types of CLP deformity. Different primary surgeons varied considerably in the percentage of their patients who ultimately required orthognathic surgery. CONCLUSION: The results of this study parallel other larger cohort studies with respect to the percentage of patients requiring orthognathic surgery. The number of prior operations does not significantly affect the later need for orthognathic surgery. PMID- 9216502 TI - Comparison of habitual masticatory cycles and muscle activity before and after orthognathic surgery. AB - PURPOSE: The purpose of this investigation was to study the long-term effects of orthognathic surgery on mastication in patients before and after four surgical procedures: mandibular advancement, maxillary intrusion, maxillary intrusion with mandibular advancement, and maxillary inferior repositioning. MATERIALS AND METHODS: The components and timing of mandibular motion, electromyography (EMG), and estimated biting forces during mastication were studied in 61 patients who underwent orthognathic surgery for correction of four different deformities. The data were statistically compared with 38 control subjects using ANOVA. RESULTS: Preoperatively, there were no significant differences in the duration of the chewing cycles and mandibular excursions among the groups, nor did surgery have any affect on these variables. Before surgery, estimated occlusal forces in the patient groups were smaller than controls. Although these appeared to increase after surgery, the increases did not exceed changes in our untreated controls. CONCLUSIONS: The results of this study suggest that, with the exception of EMG and occlusal forces, mastication in orthognathic surgery patients is not significantly different from controls either before or after surgery. EMG during mastication, although significantly lower than in controls before surgery, showed significant increases after surgery, but these increases did not bring estimated occlusal forces up to control levels. PMID- 9216503 TI - Elution of proteins by continuous temporomandibular joint arthrocentesis. AB - PURPOSE: The purpose of this study was to determine whether specific proteins recovered from human temporomandibular joints (TMJs) by superior space arthrocentesis are eluted at different outflow volumes. MATERIALS AND METHODS: Twenty subjects with unilateral TMJ pain and restricted mandibular range of motion underwent superior space arthrocentesis of the affected TMJ. Sixteen serial fractions of the arthrocentesis outflow volume were collected for analysis. The protein content of each fraction was determined by a BCA protein assay and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). In addition, samples from each collected fraction were assayed for protease activity. RESULTS: The average amount of protein recovered in the total 32 mL of collected arthrocentesis fluid was 1.5 mg (0.72 to 2.1 mg). Significant differences (P = .03) in total protein recovered from arthrocentesis fluid were observed between males (0.824 +/- 0.43 mg/20 mL) and females (1.389 +/- 0.54 mg/20 mL). In general, protein concentration declined serially in collected TMJ lavage fluid fractions. Specific proteins and proteases detected in the lavage fluid were eluted at different outflow volumes. CONCLUSIONS: Although specific proteins are eluted from the TMJ at different outflow volumes during arthrocentesis, the procedure effectively reduces the protein concentration of the lavage fluid in a volume-dependent manner. Based on empirical assumptions, it is estimated that approximately 100 mL of total arthrocentesis volume is sufficient for a therapeutic lavage of the superior joint space of the human TMJ. PMID- 9216505 TI - Surgically-assisted rapid palatal expansion for management of transverse maxillary deficiency. PMID- 9216504 TI - Reconstruction of mandibular continuity defects in dogs using poly (L-lactide) mesh and autogenic particulate cancellous bone and marrow: preliminary report. AB - PURPOSE: This study evaluated the reconstruction of continuity defects in the canine mandible using a poly [L-lactide] (PLLA) mesh tray and particulate cancellous bone and marrow (PCBM). MATERIALS AND METHODS: Eight adult dogs were divided into two groups of four dogs each. In group A, each dog had a tray fixed with stainless steel wires on each side of the mandibular stumps with the concave surface of the tray attached to the inferior border of the mandible (U-fixation). In group B, the concave surface was attached to the superior border (inverted U fixation). Each tray was filled with PCBM from the ilium. After the operation, the dogs were radiographed, and specimens were examined histologically at 3-, 6-, and 12-month intervals. RESULTS: All of group A showed good clinical healing and the continuity of the mandibular bone was regained within 3 months postoperatively. However, fibrous tissue had invaded through the area above the tray, resulting in a poorly shaped alveolar ridges. In group B, the dogs showed good bony regeneration with well-shaped alveolar ridges. However, two animals in this group had partial exposure of the PLLA mesh tray into the oral cavity. CONCLUSION: It is suggested that a combination of the PLLA mesh and PCBM grafts might be a useful technique for functional reconstruction of the jaw bone, specifically using method A (U-fixation) as a technique to reconstruct continuity defects of the mandible, and method B (inverted, U-fixation) as a promising method for alveolar reconstruction to make wearing dentures possible. PMID- 9216506 TI - Segmental LeFort I osteotomy for management of transverse maxillary deficiency. PMID- 9216507 TI - Management of injuries to the auricle. PMID- 9216508 TI - An asymptomatic enlargement of the mandible causing marked root resorption. PMID- 9216509 TI - Dissecting aneurysm of the internal carotid artery after a mandibular osteotomy. PMID- 9216510 TI - Oral sebaceous carcinoma: report of a case. PMID- 9216511 TI - Epithelioid sarcoma of the temporomandibular region: a case report. PMID- 9216512 TI - Papillary cystadenoma of the palate: a case report and ultrastructural study. PMID- 9216513 TI - Adenocarcinoma originating in the sublingual gland: report of a case. PMID- 9216514 TI - Solitary intrabony neurofibroma of the maxilla. PMID- 9216515 TI - Surgical simulation for reconstruction of mandibular bone defects using photocurable plastic skull models: report of three cases. PMID- 9216516 TI - Cutaneous paresthesia. PMID- 9216517 TI - Continuing the use hydroylapatite for ridge augmentation. PMID- 9216518 TI - The role of surgery for temporomandibular disorders. PMID- 9216519 TI - Dermatoses of the glans penis and prepuce. AB - A wide range of infectious, neoplastic, and inflammatory dermatoses can affect the glans penis or prepuce. Some are unique to the genitalia. Other more common dermatoses may have a unique appearance when they involve genital skin and mucosa. A thorough understanding of regional anatomy and a systematic diagnostic approach are helpful in the management of a refractory penile dermatosis. We review embryology and regional anatomy, drug-induced eruptions, allergic and irritant dermatitis, infection, neoplasia, and traumatic and inflammatory dermatoses as they relate to the glans and prepuce. Our discussion focuses on the clinical features, office laboratory studies, and histopathologic findings that assist in diagnosis and treatment. PMID- 9216520 TI - A novel nonepidermolytic palmoplantar keratoderma: a clinical and histopathologic study of six cases. AB - BACKGROUND: Some hereditary palmoplantar keratodermas (PPK) have been defined at the molecular level. OBJECTIVE: Our purpose was to establish the cause of a hereditary PPK with unique histopathologic findings in the epidermis. METHODS: Investigative studies included light and electron microscopy and determination of genomic DNA sequence. RESULTS: Six patients with PPK were found to have unique changes in the epidermis characterized by orthokeratosis, parakeratosis, perinuclear vacuolization, and keratohyalin granules that varied in size and shape and were located in the cell periphery. Electron microscopy showed the perinuclear region contained many ribosomes and vacuoles and was surrounded by a tonofibril shell. Family involvement suggested a dominant disorder. However, no mutation of keratin genes 1, 6a, 9, or 16 was found. CONCLUSION: The histopathologic features of this unique PPK most closely resemble Curth-Macklin ichthyosis for which the genetic basis has not been established. Further genetic studies are needed. PMID- 9216521 TI - Relation between expression of adhesion molecules and clinical behavior in cutaneous follicle center cell lymphomas. AB - BACKGROUND: Primary cutaneous follicle center cell lymphomas (PCFCCLs) of the head or trunk have a much better prognosis than morphologically similar large cell lymphomas on the legs or follicle center cell lymphomas involving the skin secondarily (SCFCCLs). Recent studies suggest a relation between the expression of adhesion molecules and clinical behavior of malignant B-cell lymphomas. OBJECTIVE: Our purpose was to investigate a potential relation between the expression of adhesion molecules and clinical behavior of these three prognostically different groups of cutaneous B-cell lymphomas. METHODS: Immunohistochemical studies with a selected panel of monoclonal antibodies against adhesion molecules were performed on 10 PCFCCLs on the head or trunk, five PCFCCLs of the legs, and seven SCFCCLs. Expression of adhesion molecules was correlated with clinical and follow-up data. RESULTS: PCFCCLs of the head and trunk expressed ICAM-1 (80%) and LFA-1 (50%) much more frequently than PCFCCLs of the legs (40% and 20%, respectively) and SCFCCLs (14% and 14%, respectively). VLA 4 was expressed in 60% of PCFCCLs of the legs, but not by the PCFCCLs of the head or trunk. Absence of both ICAM-1 and LFA-1 on the neoplastic B cells correlated with a poor prognosis (seven of nine patients died of systemic lymphoma). In contrast, none of the patients with expression of LFA-1 or ICAM-1 have died of lymphoma thus far. CONCLUSION: Our results suggest a relation between the expression of adhesion molecules and the differences in clinical behavior between different groups of primary and secondary cutaneous follicle center cell lymphomas. PMID- 9216522 TI - Development and validation of a quality of life instrument for cutaneous diseases. AB - BACKGROUND: The Dermatology-Specific Quality of Life (DSQL) instrument is a new tool to quantify the effects of skin disease on physical discomfort and symptoms, psychologic well-being, social functioning, self-care activities, performance at work or school, and self-perceptions. OBJECTIVE: Our purpose was to describe the reliability and validity of the DSQL in two disease cohorts comprising patients with contact dermatitis and acne vulgaris. METHODS: Reliability was assessed from the internal consistency of the items, and correlations were made between DSQL scores from a 3- to 7-day retest. Validity was assessed from correlations of DSQL scales with global ratings of bothersome symptoms and perceived severity and by discrimination of clinically defined severity groups. RESULTS: The DSQL scales had high internal consistency (0.70 to > 0.90) and test-retest reliability (r = 0.81 to 0.89), and were moderately to highly correlated with patient global ratings of symptom distress (r = 0.25 to 0.67) and overall disease severity (r = 0.19 to 0.54). Patients rated with severe contact dermatitis or acne scarring had higher DSQL scores than those with less severe skin disease. Factor analyses found separate dimensions of physical, emotional, and social functioning involvement from skin disease. CONCLUSION: The DSQL provides valid and reliable assessments of quality of life impacts associated with acne and contact dermatitis. PMID- 9216523 TI - A comparison of subjective and objective measures of reduction of psoriasis with the use of ultrasound, reflectance colorimetry, computerized video image analysis, and nitric oxide production. AB - BACKGROUND: Studies of antipsoriatic therapy often rely on subjective scoring. Objective measures have been developed but have not previously been compared with subjective scoring. OBJECTIVE: Our purpose was to compare subjective and objective measures of reduction of psoriasis with topical therapy. METHODS: A 2 week, double-blind, left-to-right comparative trial of betamethasone valerate against white soft paraffin was performed in 12 patients. The subjective scores were erythema, elevation, scale, and a composite total. Objective measures were nitric oxide production measured by chemiluminescence; erythema reflectance; ultrasound scan for thickness, scale, and echo-poor zone; and computerized image analysis of video images. RESULTS: Subjective and objective measures had similar power to detect therapeutic effect. The subjective measures showed greater variation and relatively overestimated improvement. There was correlation between measures and estimates for area, redness, and thickness. Nitric oxide production was the most powerful objective measure. CONCLUSION: Thickness determined by ultrasound scan and nitric oxide production are useful measures of reduction of psoriasis, which lend themselves to more powerful statistical tests than subjective interval data. PMID- 9216525 TI - Epiluminescence microscopy: criteria of cutaneous melanoma progression. AB - BACKGROUND: Cutaneous melanoma develops through a series of evolutionary steps (intraepidermal, radial, and vertical growth phases) that are traceable in specific histologic features. Epiluminescence microscopy (ELM) is an in vivo technique that enables the visualization of morphologic structures in pigmented lesions correlated with specific histologic architectural characteristics. Many ELM criteria associated with cutaneous melanoma have been described, but their correlation with tumor progression has not yet been established. OBJECTIVE: In this preliminary study our purpose was to explore the possibility of recognizing ELM criteria that allow the in vivo detection of the various phases of melanoma progression as well as tumor depth. METHODS: Seventy-two cutaneous melanomas (41 "thin" melanomas [TnM], < 0.76 mm thickness, and 31 "thick" melanomas [TkM], > 0.75 mm thickness) were investigated with ELM for the presence of nine standard ELM criteria; their significance was determined by calculating the chi-square test of independence. RESULTS: A significant association is found between the presence of pigment network and TnM and between the presence of gray-blue areas, vascular pattern, and TkM. Moreover, pigment network plus radial streaming is the most significant association of ELM criteria in TnM, whereas gray-blue areas plus vascular pattern is the greatest in TkM. CONCLUSION: This study shows a good correlation between certain ELM criteria and the histologic architecture of cutaneous melanoma for a preoperative evaluation of the tumor thickness. Further investigation is needed for verifying on a larger number of cases our pilot estimates of sensitivity and specificity of ELM criteria in thin and thick melanomas. PMID- 9216524 TI - Prognostic factors and evaluation of mycosis fungoides and Sezary syndrome. AB - BACKGROUND: Staging evaluations of patients with mycosis fungoides (MF) and Sezary syndrome (SS) are performed to individualize therapy and to predict survival. OBJECTIVE: Our purpose was to determine the prognostic factors in patients with MF and SS. METHODS: A retrospective study of 101 patients was performed. For inclusion in the study, patients had to have been evaluated for MF or SS within 6 months of the initial definitive histologic diagnosis. The evaluation included physical examination, chest radiograph, peripheral blood smear, lymph node biopsy, bone marrow biopsy, gallium 67 scan, liver-spleen scan and computed tomography (CT) of the chest, abdomen, and pelvis. RESULTS: The type of skin disease present at initial diagnosis was a good prognostic indicator of survival and clinical outcome. Univariate adverse prognostic features included hepatosplenomegaly or adenopathy by CT scan, abnormal liver-spleen scan, abnormal gallium scan, adenopathy, and peripheral blood, bone marrow, and lymph node involvement. Independent prognostic factors in multivariate analysis were the type of skin involvement as well as peripheral blood and visceral involvement. CONCLUSION: Our study confirms previous reports that type of skin and peripheral blood and visceral involvement are important prognostic factors in patients with MF or SS. Our results support the finding that patients with T1 stage disease have an excellent survival outlook and clinical outcome. PMID- 9216526 TI - Pulsed laser treatment in children and the use of anesthesia. AB - BACKGROUND: The rationale for choosing certain anesthetic options in children when they are being treated with pulsed lasers is unclear. OBJECTIVE: Our purpose was to assess the safety and side effects of general anesthesia in the treatment of vascular lesions and to compare this to treatment outcome in the office setting. METHODS: We carried out a retrospective chart review of 179 patients, with an age range of 5 weeks to 18 years, who received laser treatment and underwent different anesthetic modalities. The age of the patient and the size, location, and severity of the vascular lesion were also noted. RESULTS: The factors determining the type of anesthesia to use included (1) the age of the patient, (2) the number of treatments, and (3) the size and location of the lesion. Our data showed minimal risk and sequelae of general anesthesia in the treatment of vascular lesions in children. CONCLUSION: Proper selection of anesthesia is a key factor in dealing with children. Office surgery can be performed safely when small lesions are treated. The use of general anesthesia in the treatment of port-wine stains in children does not appear to be accompanied by increased risk. PMID- 9216527 TI - Recurrent basal cell carcinoma treated with cryosurgery. AB - BACKGROUND: Although there are reports of cure rates achieved by cryosurgery for primary basal cell carcinomas (BCCs), there are few data on the cryosurgical treatment of recurrent BCCs. OBJECTIVE: Our purpose was to discuss case selection, cryosurgical management, and results of therapy. METHODS: Cryosurgery was performed in 54 patients with 56 recurrent BCCs. The treatment consisted of aggressive freezing including an adequate margin of surrounding tissue. RESULTS: Wound healing was favorable and the cosmetic results were excellent. Two recurrences were found and were referred for Mohs micrographic surgery. CONCLUSION: We conclude that cryosurgical treatment of selected recurrent BCCs yields results that compare favorably with other methods of treatment. PMID- 9216529 TI - Interactions between calcipotriene and ultraviolet light. AB - BACKGROUND: Calcipotriene is often used with UVB or PUVA, but interactions between UV radiation and calcipotriene have not been examined extensively. OBJECTIVE: Our purpose was to examine interactions between calcipotriene and UV light. METHODS: Minimal erythema doses (MEDs) were determined with UVB and immediate pigment darkening was measured for UVA. The effect of calcipotriene ointment applied before phototesting was examined. Thick and thin applications of calcipotrience were compared. Calcipotriene ointment was applied to a small area on the skin before phototherapy. Patients received either UVB, PUVA, UVA, or no phototherapy. After phototherapy, the ointment was collected and assayed by reverse-phase, high-performance liquid chromatography. RESULTS: MEDs for UVB and immediate pigment darkening for UVA were unaffected by calcipotriene. Thick application of calcipotriene, however, increased the MED, UVA caused substantial reductions in the concentration of detectable calcipotriene. CONCLUSION: When used in conjunction with PUVA, calcipotriene should be applied after exposure to UVA. PMID- 9216528 TI - Tazarotene gel, a new retinoid, for topical therapy of psoriasis: vehicle controlled study of safety, efficacy, and duration of therapeutic effect. AB - BACKGROUND: Topical therapy providing initial improvement and maintenance of effect after treatment of the large majority of patients with limited, mild to moderate psoriasis is not presently available. Previous topical retinoids have generally been either ineffective or too irritating for therapy of psoriasis. OBJECTIVE: Our purpose was to evaluate a new topical retinoid, tazarotene, in the treatment of stable plaque psoriasis during treatment and posttreatment periods. METHODS: In a double-blind manner, 324 patients were randomly selected to receive tazarotene 0.1% or 0.05% gel, or vehicle control, once daily for 12 weeks and were then followed up for 12 weeks after treatment. RESULTS: Of the total, 318 patients could be evaluated. Tazarotene gels were superior (p < 0.05) to vehicle, often as early as treatment week 1, in all efficacy measures: plaque elevation, scaling, and erythema; treatment response; percentage treatment success (patients with > or = 50% improvement); and time to initial success. Efficacy was equivalent on target lesion sites (trunk or limbs and knees or elbows) and overall. A sustained therapeutic effect was observed for 12 weeks after treatment. Tazarotene gel was cosmetically acceptable. There was low systemic absorption, limiting toxicity to local irritation. CONCLUSION: Once-daily tazarotene was effective and safe as a topical monotherapy for plaque psoriasis, providing rapid reduction of signs and symptoms. PMID- 9216531 TI - Eosinophils in fibrous tracts and near hair bulbs: a helpful diagnostic feature of alopecia areata. AB - BACKGROUND: When biopsy specimens lack a "swarm of bees" peribulbar lymphoid infiltrate, the diagnosis of alopecia areata depends on the recognition of other histologic features of the disease. OBJECTIVE: Our objective was to determine the frequency of the presence of eosinophils in biopsy specimens of alopecia area'a in relation to the other major histologic features of the disease. METHODS: Biopsy specimens from 71 patients with alopecia areata were studied. RESULTS: Eosinophils were present in 38 of the 71 cases. A peribulbar lymphoid infiltrate was absent in 27 of the 71 cases. Eosinophils were present in 13 of these cases. Multiple catagen hairs and pigment casts (features which may lead to confusion with trichotillomania) were present in 39 cases. The presence of eosinophils was found to be a helpful diagnostic feature in cases with potential for misdiagnoses as trichotillomania. CONCLUSION: Eosinophils are common in all stages of alopecia areata, both within the peribulbar infiltrate and within fibrous tracts. A "swarm of bees" peribulbar infiltrate may be absent. The presence of eosinophils is a helpful diagnostic feature of alopecia areata. PMID- 9216530 TI - The use of EMLA cream and 1% lidocaine infiltration in men for relief of pain associated with the removal of genital warts by cryotherapy. AB - BACKGROUND: Surgical procedures used to remove genital warts (cryotherapy, electrodesiccation) are painful. Attempts to reduce the discomfort of surgery by prior lidocaine infiltration anesthesia are compromised by the pain of the infiltration. OBJECTIVE: Our purpose was to determine the efficacy of topically applied lidocaine/prilocaine cream to reduce the pain of lidocaine infiltration and the pain associated with cryotherapy to remove genital warts. METHODS: Men, scheduled for removal of genital warts by cryotherapy, were randomly selected to receive one of three treatments: (1) lidocaine/prilocaine cream application, (2) 1% lidocaine infiltration, and (3) lidocaine/prilocaine cream application followed by infiltration of 1% lidocaine. RESULTS: Application of lidocaine/prilocaine cream for 15 minutes markedly reduced the pain of lidocaine infiltration. The combination of lidocaine/prilocaine cream followed by infiltration of 1% lidocaine gave greater pain relief from the cryotherapy than did either anesthetic alone. CONCLUSION: The application of lidocaine/prilocaine cream as an adjunct to lidocaine infiltration reduced the pain of infiltration and the pain associated with cryotherapy for the removal of genital warts. PMID- 9216532 TI - Kaposi's sarcoma-associated herpesvirus/human herpesvirus-8: a new virus in human pathology. AB - The discovery of a new human herpesvirus in Kaposi's sarcoma (KS) tissue of patients with AIDS has opened up new vistas in virology and oncology. This herpesvirus was first descriptively named KS-associated herpesvirus (KSHV), but was recently renamed human herpesvirus 8 (HHV8). KSHV/HHV8 DNA has been found in all forms of KS, suggesting that it might be involved in the pathogenesis of KS. In addition, KSHV/HHV8 can be detected in both malignant and benign lymphoproliferative disease. KSHV/HHV8 was also found in patients with angiosarcoma of the face and angiolymphoid hyperplasia with eosinophilia. Although only a limited portion of the virus has been sequenced, KSHV/HHV8 is equipped with genes that could confer oncogenic potential. The virus can now be cultured, providing the possibility for studies of viral replication and the mode of transmission. The recently developed serologic assays for antiviral antibodies suggest that infection with KSHV/HHV8 is not ubiquitous because KSHV/HHV8 seropositivity is limited to a small proportion of the population. PMID- 9216533 TI - Surgical pearl: easy and (usually) painless removal of adhesive dressings. PMID- 9216534 TI - Granulomatous-ulcerative vulvar cryptococcosis in a patient with advanced HIV disease. PMID- 9216535 TI - Injection site vasculitis in a patient receiving interferon alfa for chronic hepatitis C. PMID- 9216537 TI - Sweet's syndrome associated with Helicobacter pylori infection. PMID- 9216536 TI - D-penicillamine-induced pemphigus foliaceus with autoantibodies to desmoglein-1 in a patient with mixed connective tissue disease. PMID- 9216538 TI - Localized cutaneous amyloidosis associated with mycosis fungoides. PMID- 9216539 TI - Papular mucinosis associated with AIDS: response to isotretinoin. PMID- 9216540 TI - Generalized cutaneous metastases from breast adenocarcinoma. PMID- 9216542 TI - Acne lesion counts and global assessments. PMID- 9216541 TI - Cutaneous nodular reaction to oral mercury. PMID- 9216543 TI - Sign of Leser-Trelat. PMID- 9216544 TI - Irritant reactivity in noncutaneous atopy. PMID- 9216545 TI - Leishmaniasis. PMID- 9216546 TI - Messages for the masses: food and nutrition issues on television. PMID- 9216547 TI - Combating obesity in the United States. PMID- 9216548 TI - Dual Medicare-Medicaid eligibility spurs health care reform. PMID- 9216549 TI - Dietetics and nutrition: impact of scientific advances and development. PMID- 9216550 TI - Population nutrient intake approaches dietary recommendations: 1991 to 1995 Framingham Nutrition Studies. AB - OBJECTIVE: To estimate population nutrient intake levels and to assess adherence to current dietary recommendations for health promotion and disease prevention. DESIGN: Cross-sectional analysis of nutrient intake estimated from 3-day food records. Median macronutrient and micronutrient intake levels for men, women, and the total population are reported along with the proportions of men and women who achieved intakes compatible with nutrient goals defined by published guidelines. SETTING: Adult participants (2,520: 1,375 women and 1,145 men) in the Framingham Offspring-Spouse Study surveyed between 1991 and 1995. STATISTICAL ANALYSES: chi 2 Analyses were used to test for gender differences in the proportions of persons who had intakes that met nutrient guidelines. RESULTS: Population intake levels of certain key nutrients, including total and saturated fat, appear to be approaching recommended levels. High proportions of the Framingham population (70% or more) met current recommendations for intakes of protein, polyunsaturated and monounsaturated fat, cholesterol, alcohol, vitamins C and B-12, and folacin. About half or fewer met guidelines for carbohydrate; total and saturated fat; fiber; beta carotene; vitamins A, E, and B-6; calcium; and sodium. Important gender differences in the proportion of those meeting nutrient guidelines were observed for 12 of the 18 nutrients examined, including carbohydrate; total, saturated, and monounsaturated fat; cholesterol; fiber; sodium; calcium; and several vitamins. CONCLUSIONS: Although progress has been made toward achieving population adherence to preventive nutrition recommendations, large proportions of adults fall short of guidelines for some key nutrients. Differences in adherence rates between men and women suggest areas for gender-specific, targeted nutrition messages and behavioral interventions. PMID- 9216551 TI - Food preferences predict eating behavior of very young Mohawk children. AB - OBJECTIVE: To collect baseline data on energy and nutrient intake and nutrition knowledge, attitudes, and behavior of very young Mohawk children to assist the community in planning an appropriate, targeted nutrition and exercise intervention. DESIGN: Energy and nutrient intake data were collected from 24-hour recalls conducted in the children's homes. Nutrition knowledge, attitudes, and behavior were assessed using a 38-item questionnaire that asked children to report on what foods they like the best, eat most of the time, and think are healthful. The questionnaire was completed in an elementary school on the reservation. Before data collection, we hypothesized that the average diet of the Mohawk children would not meet national dietary recommendations. SUBJECTS: One hundred forty-three children, prekindergarten through third grade (aged 4 to 9 years), completed the 24-hour recalls and the questionnaire. An additional 136 children, also prekindergarten through third grade, completed the questionnaire (n = 279). STATISTICS: Analysis of variance with a Scheffe's multiple-comparison test was used to test for differences among grades and genders for energy and nutrient intake and questionnaire scores. Multiple regression analysis was used to assess the relationship between eating behavior and selected variables. RESULTS: A mean daily energy intake of 1,980 kcal consisted of 34% fat, 13% protein, and 52% carbohydrate with 13 g fiber and 235 mg cholesterol. Food preferences were the strongest predictor of behavior, they explained 71% of the variation in the behavior score. APPLICATIONS: The major finding of this study, that food preferences are the strongest predictor of reported eating behavior in very young Mohawk children, has implications for behavior change interventions. Focusing on changing what children like to eat, through repeated exposure to new foods in a positive social context, is more likely to change what foods they choose than is simple nutrition education. PMID- 9216552 TI - Among young adults, college students and graduates practiced more healthful habits and made more healthful food choices than did nonstudents. AB - OBJECTIVES: Health-related characteristics and habits and food choices of young adults were compared for three groups: college students, college graduates, and nonstudents. DESIGN: Subjects completed a mailed survey that included questions about demographics, attitudes, and behaviors and a food frequency questionnaire. Main outcome measures were health-related characteristics and habits and food choices. SUBJECTS: Female (n = 758) and male (n = 580) 18- to 24-year-olds in nine states who were selected randomly by zip code in each state. The response rate averaged 43% for all states. STATISTICAL ANALYSES PERFORMED: Analysis of variance of chi 2 tests were applied to health-related personal characteristic variables and the Kruskal-Wallis analysis of variance was applied to food consumption variables for women and men separately. RESULTS: According to self reported heights and weights, female nonstudents were more often overweight than female students or graduates. Nonstudents of both genders reported smoking more often than students or graduates. College students and graduates ate more grain foods high in dietary fiber, more fruits and dark-green vegetables, and more lower-fat milk and meats than nonstudents. CONCLUSIONS/APPLICATIONS: Nonstudents were at greater health risk for some chronic illnesses, because of poorer health habits and food choices, than were college students and graduates. The behavior of nonstudents implies weaker response to messages promoting weight control, smoking cessation, and observance of the Dietary Guidelines for Americans than behavior exhibited by students or college graduates. Health promotion efforts could be enhanced by identifying demographic, educational, situational, and formative influences on positive health and dietary behaviors of young adults. PMID- 9216553 TI - Nutrition Screening Initiative Checklist may be a better awareness/educational tool than a screening one. AB - OBJECTIVE: To evaluate the Nutrition Screening Initiative (NSI) checklist as a screening and an awareness/educational tool in an elderly population. DESIGN: Epidemiologic follow-up study. Information similar to the questions of the NSI checklist was collected by the Nutrition Status Survey of Boston elders between 1981 and 1984. Vital status of volunteers was obtained during 8 to 12 years of follow-up. SUBJECTS/SETTING: Community-dwelling men (n = 200) and women (n = 381) aged 60 years and older who participated in the survey. STATISTICAL ANALYSES PERFORMED: Multivariate analysis was used to assess the association between mortality and each of the NSI-similar questions and the cumulative score, which is the sum of the values assigned to each question. Attributable risk percent, a measure of association, was calculated to measure the percentage of deaths that could potentially be prevented if the risk factors or their consequences were eliminated. RESULTS: Eating meals alone, problems biting or chewing, difficulties with shopping or cooking, and taking more than three medications per day were positively associated with mortality (P < .05). The cumulative score, although significant, was a weaker predictor of mortality. Attributable risk percent of mortality was 19.9% and 51.2% for men and women, respectively. APPLICATIONS/CONCLUSIONS: Some but not all of the individual questions of the NSI checklist equivalent were significantly associated with mortality and identify specific problems that may have a long-term negative effect yet may be missed if the cumulative score were the sole criterion for screening people. The attributable risk percent suggests that the checklist may be best used as an awareness/educational tool as intended originally and could have an important public health effect on early death of community-dwelling elderly people. PMID- 9216554 TI - Creatine supplementation enhances muscular performance during high-intensity resistance exercise. AB - OBJECTIVE: This study was undertaken to investigate the influence of oral supplementation with creatine monohydrate on muscular performance during repeated sets of high-intensity resistance exercise. SUBJECTS/DESIGN: Fourteen active men were randomly assigned in a double-blind fashion to either a creatine group (n = 7) or a placebo group (n = 7). Both groups performed a bench press exercise protocol (5 sets to failure using each subject's predetermined 10-repetition maximum) and a jump squat exercise protocol (5 sets of 10 repetitions using 30% of each subject's 1-repetition maximum squat) on three different occasions (T1, T2, and T3) separated by 6 days. INTERVENTION: Before T1, both groups received no supplementation. From T1 to T2, both groups ingested placebo capsules. From T2 to T3, the creatine group ingested 25 g creatine monohydrate per day, and the placebo group ingested an equivalent amount of placebo. MAIN OUTCOME MEASURES: Total repetitions for each set of bench presses and peak power output for each set of jump squats were determined. Other measures included assessment of diet, body mass, skinfold thickness, and preexercise and 5-minute postexercise lactate concentrations. RESULTS: Lifting performance was not altered for either exercise protocol after ingestion of the placebos. Creatine supplementation resulted in a significant improvement in peak power output during all 5 sets of jump squats and a significant improvement in repetitions during all 5 sets of bench presses. After creatine supplementation, postexercise lactate concentrations were significantly higher after the bench press but not the jump squat. A significant increase in body mass of 1.4 kg (range = 0.0 to 2.7 kg) was observed after creatine ingestion. CONCLUSION: One week of creatine supplementation (25 g/day) enhances muscular performance during repeated sets of bench press and jump squat exercise. PMID- 9216555 TI - Common health problems among minority elders. AB - This article reviews the primary health problems of African-American, Hispanic American, Asian/Pacific Islander-American, and Native-American elders. The goal is to familiarize practicing dietitians with the differences in longevity, disease spectrum, and functional status (where data are available) for each of these ethnic groups. These data should be of assistance in making decisions regarding dietary counseling for ethnic elders. It is acknowledged that most data accumulated according to race do not accurately measure ethnicity. The degree of acculturation may vary widely among individuals. Therefore, it is recommended that dietitians solicit clients' perceptions of the factors that may contribute to illness and the barriers to implementing recommended remedies. PMID- 9216556 TI - Twenty years of WIC: a review of some effects of the program. AB - The Special Supplemental Nutrition Program for Women, Infants, and Children (WIC) began in 1974 after a 2-year pilot program. WIC links food assistance and nutrition education to health care for at-risk persons. The program had approximately 344,000 participants in 1975 and has grown to provide services to nearly 6 million participants. Infants born to women who participate in WIC during pregnancy tend to have a slightly higher mean birth weight than those born to women who were eligible but did not participate in WIC. Higher birth weight has been associated with a slightly higher mean gestational age. The prevalence of low birth weight and very low birth weight among infants and the prevalence of iron deficiency anemia among toddlers and preschool children is lower for those participating in WIC than for those not participating in WIC. PMID- 9216557 TI - Functions of dietitians providing nutrition support to patients with inherited metabolic disorders. AB - This article examines functions of dietitians who provide nutrition services to patients with inherited metabolic disorders. A survey questionnaire was developed and pilot-tested in a sample of dietitians, revised, and mailed to all dietitians in the United States who treat patients with inherited metabolic disorders. One hundred forty-two usable questionnaires were returned. Descriptive statistics were used to calculate response frequency. The highest academic degree attained by 37% of the dietitians was a bachelor's degree; 58% had earned a master's degree and 5% a doctorate. Dietitians provided nutrition services during diagnosis, critical illness, and long-term care. More than 90% of the dietitians performed these functions: evaluated nutrition status; prepared, implemented, and evaluated the nutritional support plan; revised the nutrition support plan as needed; monitored dietary compliance; coordinated care with other agencies; developed materials and educated parents, caregivers, and patients about the nutrition support plan; and recorded information in the medical record. Without nutrition support, patients with inherited metabolic disorders may become mentally retarded, experience neurologic or metabolic crises, or die. PMID- 9216558 TI - Profile of American-trained dietitians in the international setting: survey of the members of the American Overseas Dietetic Association. PMID- 9216559 TI - Honest but invalid: what subjects say about recording their food intake. PMID- 9216560 TI - 'We think your son has Lennox-Gastaut syndrome'--a case study of monosodium glutamate's possible effect on a child. PMID- 9216561 TI - The challenges of assessing fat intake in cancer research investigations. AB - Ecologic comparison of the incidence of cancer (eg, largescale differences between countries in the incidence of breast, prostate, and colon cancer) can be explained best by substantial differences in the intake of dietary fat. Additionally, there is a vast amount of animal and mechanistic data that strongly supports the hypothesis that ditary fat, independent of caloric intake, appears to have a major effect on the incidence and mortality rates for cancer. Yet, results from human case and cohort studies are inconsistent in linking carcinogenesis with fat intake. This is due to several factors. Reported intakes may not reflect previous long-term intakes and may be con-founded by several sources of error, including memory and estimates of portion size. Additionally, ongoing media reports of adverse health effects from high-fat diets may impart a social desirability bias to self-reporting of fat intake. These factors may be significant when investigating the relationship between dietary intake and cancer. Studies have shown considerable error in self-reported dietary data, with under-estimations in energy intake ranging from 3% to 18%. Such a wide range likely is due to differences in dietary assessment methodologies, which highlights the need to continue to develop improved techniques of data collection to relate nutrition better to health outcomes. The Women's Intervention Nutrition Study (WINS) is investigating the effect of dietary fat on the incidence of recurrence and survival in women with early-stage breast cancer. WINS is employing the multiple-pass 24-hour telephone recall system along with enhanced quality control measures to assess dietary intake. This dietary assessment method is particularly applicable when comparing two populations when one population is treated by an extensive dietary intervention. PMID- 9216562 TI - Dietary fat and chronic diseases: epidemiologic overview. AB - The association between dietary fat consumption and risk of cancer, especially colon, breast, prostate, and ovary cancer, has been debated for many years. Ecologic studies over the past 30 years have demonstrated the correlation of greater dietary fat intake with higher mortality due to various cancers. Migrant studies also have shown that increased fat consumption may be associated with increased risk of cancer. Specific saturated fatty acids raise blood cholesterol levels and, thereby, increase the risk of atherosclerosis. Greater fat, intake is a major cause of obesity and hypertension, diabetes, and gallbladder disease. Higher fat intake may heighten the risk of breast cancer directly through increased blood estrogen levels and/or secondarily through increased obesity. The critical experimental studies to determine the effects of a low-fat diet on disease risk have not been completed, but reducing fat in the US diet has the potential to decrease morbidity and mortality substantially. PMID- 9216563 TI - Dietary fat and risk of chronic disease: mechanistic insights from experimental studies. AB - The primary nutritionally linked diseases are coronary heart disease, stroke and cancers of the stomach, colon, pancreas, prostate, breast, ovary, and endometrium. Dietary fats operate through a promoting mechanism. An S-shaped dose response curve with a threshold has been demonstrated in models of breast and colon cancer in which the standard Western fat intake of 40% of energy yields a high level of promotion, and reduction of fat to 10% to 20% of energy (the traditional Japanese fat intake) has a low promoting action. In models of breast and colon cancer, saturated fats such as beef fat or lard, and monounsaturated oils, such as olive oil, display only a weak promoting effect, with the incidence of induced tumors being similar at intake levels of 40% and 10% of energy. On the other hand, the n-6-polyunsaturated oils display a strong promoting effect. Such findings may have a parallel in the low but definitely increasing slope of postmenopausal breast cancer incidence in the past 30 years as the American public decreased saturated fat intake to avoid heart disease and increased use of the n-6-polyunsaturated oils. Mechanisms underlying the cancer-promoting effect in the colon stem from increased hepatic production of bile acids, which are transferred to the intestinal tract via the bile. Ingestion of 40% fat calories yields higher concentrations of bile acids in the colon than lower levels of dietary fat ingestion. Cancer in the mammary gland is promoted through higher concentrations of fats and phospholipids in the gland as well as increased levels of estrogen secondary to production by the ovary and other endocrine tissues that, in turn, affect the generation of pituitary hormones such as prolactin and growth hormone. The n-3-fats, as found in fish and fish oils, have a pronounced inhibitory effect in models of colon and breast cancer, presumably through their shifting of prostaglandin metabolism to the generation of prostaglandins, which lower cell proliferation potential and, thus, decrease promotional effects. The role of dietary fat as a promoter can be modified by other nutritional components. Finally, one of the best pieces of evidence for an enhancing effect of many dietary fats in the nutritionally linked cancers is the current increase in the incidence of these diseases in Japan as the nutritional habits of people in that country become more Westernized. PMID- 9216564 TI - Fat, caloric intake, and obesity: lifestyle risk factors for breast cancer. AB - Dietary fat is a likely important determinant of postmenopausal breast cancer as part of an intricate and inseparable interaction of lifestyle cancer risk factors that include dietary fat, type of fat, energy intake and expenditure, and obesity. These factors possibly build upon individual susceptibilities derived from a complex array of polygenetic risk determinants. Epidemiologic studies have not provided conclusive evidence for a dietary fat-breast cancer association, partly because studies that focus on a single nutrient cannot always evaluate readily the interactive effects of other lifestyle factors. Further, persons generally underestimate their usual dietary intake, measured by either food frequency questionnaires (FFQs) or diet records. A dietary measurement model that accounts for this underreporting demonstrated that FFQs and diet records may not be able to detect a dietary fat-breast cancer association because of measurement error biases. Although meta-analysis of epidemiologic data across individual studies suggests only a week association between breast cancer and dietary fat, this result is compatible with the dietary measurement model and does not rule out a contributing role for dietary fat, either alone or with other causative factors. Research is needed that focuses on a comprehensive approach to dietary lifestyle choices and breast cancer risk and that emphasizes a fat-caloric intake obesity linkage. The best hope for a definitive answer may rest with randomized, controlled clinical trials. Two such trials, the Women's Health Initiative and the Women's Intervention Nutrition Study, are under way. PMID- 9216565 TI - The effect of dietary fat, antioxidants, and pro-oxidants on blood lipids, lipoproteins, and atherosclerosis. AB - A number of primary and secondary prevention trials, including angiographic studies, have indicated that a decrease in dietary saturated fat and cholesterol produces a decrease in the blood levels of cholesterol and low-density lipoprotein (LDL) cholesterol, leading to a decrease in coronary artery disease (CAD). Increasing evidence indicates that the oxidation of LDL in human beings is atherogenic. Of the three major antioxidants, vitamin E, beta carotene, and vitamin C, the evidence is strongest that vitamin E (at a minimum dose of 100 IU/day) has a strong and independent inverse association with CAD. Selenium and flavonoids also have antioxidant properties, but their association with CAD in human beings is equivocal. Two prooxidants, homocysteine and iron, have been found to be associated with CAD. Blood homocysteine levels can be lowered significantly by an increase in dietary folic acid. Clinical trials are needed to assess expeditiously the effect of antioxidants, particularly vitamin E, and of folic acid on CAD and atherosclerosis. The substitution of monounsaturated fat for saturated fat lowers LDL and makes it less susceptible to oxidation without decreasing high-density lipoprotein (HDL) cholesterol. Studies in transgenic mice indicate that apolipoprotein A-I, the major protein of HDL, may inhibit the oxidation of LDL. Dietary trans fatty acids at the level consumed by many Americans can increase LDL cholesterol and may decrease HDL cholesterol. Individuals who have CAD or have family members who have premature CAD have delayed clearance of dietary fat, as judged by studies of postprandial triglyceride metabolism. The importance of decreasing dietary saturated fat and cholesterol is well established, but a number of other factors appear to influence the risk of CAD significantly and provide important areas for future investigation to improve prevention and treatment through better nutrition. PMID- 9216566 TI - Dietary fat and human obesity. AB - When energy is in excess, the human body processes nutrients according to an oxidative hierarchy. Excessive carbohydrate and protein intakes are disposed of by increased oxidation. In contrast, excess fat intake does not promote its own oxidation in the short- and mid-term. This leads, in the long-term, to an increase in fat stores. Although increased adiposity represents the common response to increased fat intake, there are interindividual differences in lipid oxidation (probably genetically determined) that may protect from or predispose to obesity. PMID- 9216567 TI - Cardiovascular health risks related to overweight. AB - Cross-sectional surveys of the civilian noninstitutionalized population of the United States, including in-home interviews and clinical examinations, were employed to examine trends in consumption of energy and fat, prevalence of overweight in the population, the association of overweight with levels of blood pressure and blood cholesterol, and the prevalence of high blood pressure and high blood cholesterol among the overweight compared with the nonoverweight. Data from participants 20 years of age and older are reported. Study results suggest that total mean energy intake, although generally accepted to be underreported in dietary surveys, may have increased. Total fat and saturated fat intake as a percent of energy decreased, but remained above recommended levels. Overweight has increased in the population, despite decreases in the prevalence of high blood pressure and high blood cholesterol levels. Increased levels of overweight, reported as body mass index, are associated with increased cardiovascular risk factors of high blood pressure and high blood cholesterol. These data suggest the need for health care practitioners to emphasize the requirement for energy balance (or weight loss if overweight, ie, not at a "healthy weight"). A focus on fat intake alone without emphasis on energy balance is inadequate for achieving and maintaining recommended weight. PMID- 9216568 TI - Steps toward a fat-free future. AB - The most fundamental step in discussions about reducing fat in our diets is to emphasize a patient priority. With such a philosophy, we can take actions that best serve patients, while reducing the rising costs of care and making that care more affordable. A patient priority places the focus of efforts on empowering people and giving them the best value for their health dollars. It provides consumers with choices and information about those choices. PMID- 9216569 TI - From the Miocene to olestra: a historical perspective on fat consumption. AB - Given the extraordinary dietary and geographic diversity of Pleistocene hominids, there is no single "Paleolithic diet" or average pre-Holocene fat intake. Even the Neanderthals initially were scavengers, possibly becoming seasonal hunters of large game at a later period. Fat intakes of greater than 20 g/day (11% of total caloric intake) developed after the domestication of mammals and then by selective breeding of genetically fatter animals in suitably temperate climates. By the late 1940s, the percent of fat in the diet rose to more than 40% in many Western countries (including France), decreasing somewhat to about 35% by the late 1980s in the United States, following reduced consumption of whole milk, fried meats, and other high-fat foods. Overall, fat reductions to less than 30% may be facilitated by no-fat or low-fat substitutes or texturizers or (perhaps more effectively) by increased intakes of fiber and calcium and greater reliance on fats that are poorly absorbed because of their stearate content. PMID- 9216570 TI - Why do we like fat? AB - Dietary choices are strongly influenced by the taste and texture of foods. Fats are responsible for the sensory properties of many foods and greatly contribute to eating pleasure. Although diets rich in fats tend to be more flavorful and varied, they also are high in energy. Because excessive fat consumption has been associated with higher rates of obesity and coronary heart disease, nutrition education efforts have focused on replacing dietary fats with grains, vegetables, and fruit. However, preference for high-fat foods appear to be a universal human trait, and in the absence of efficient physiologic mechanisms regulating fat intake, fat consumption appears to be determined simply by the amount of fat available in the food supply. Fat consumption at national levels is determined largely by economic variables such as urbanization or income. The question is whether these barriers can be surmounted by appropriate nutrition education and intervention programs. PMID- 9216571 TI - Fat as a risk factor for overconsumption: satiation, satiety, and patterns of eating. AB - Many people experience great difficulty in preventing energy intake from outstripping energy expenditure. Eating high-fat foods can facilitate the development of short-term positive energy balances by influencing satiation and satiety, the processes that control the size of eating episodes and the strength of postingestive appetite inhibition, respectively. An important feature of these processes is the relative potency of orosensory, postingestive (preabsorptive), and postabsorptive signals. Foods high in dietary fat have a weak effect on satiation, which leads to a form of passive overconsumption, and a disproportionately weak effect on satiety (joule-for-joule compared with protein and carbohydrate). This overconsumption (high-fat hyperphagia) is dependent upon both the high energy density and the potent sensory qualities (high palatability) of high-fat foods. A positive fat balance does not appear to generate a tendency for behavioral compensation, and there appears to be almost no autoregulatory link between fat oxidation and fat intake. The Leeds High Fat Study has found a higher frequency of obesity among high-fat than low-fat consumers, but the relationship between fat consumption and obesity is not a biologic imperative: analysis of the pathways between daily fat intakes and patterns of eating has revealed high-risk eating episodes. The physiologic responses to fat ingestion appear to be weak compared with the potent orosensory properties of high-fat foods, and such responses cannot prevent overconsumption. A first stage in a health program should be to prevent passive overconsumption. PMID- 9216572 TI - One size fits all: implications for assessing dietary behavior. AB - Accurate assessment of dietary behavior is central to the design, implementation, and evaluation of intervention programs aimed at behavior change, and use of an Eating Behaviors Questionnaire (EBQ) has been suggested for measuring dimensions of dietary fat behavior. The EBQ has proven useful in characterizing fat-related dietary patterns among middle-class, highly educated, highly motivated white women. To investigate the generalizability of the instrument, we provide findings from a community-based sample of 235 African-Americans in Maywood, Illinois, a middle-class working community outside Chicago. The sample consisted of 159 women and 76 men with an average age of 47.4 +/- 13.8 years for women and 48.1 +/- 12.1 years for men (mean +/- standard deviation; range, 18 to 87 years). The EBQ is based on four broad behavioral domains (ie, avoidance, modification, substitution, and replacement) associated with fat-related eating patterns. These behavioral domains are composed of specific dietary behaviors (factors). Using a scoring system that allowed all participants to be included in all analyses, we identified a set of factors characterizing eating patterns in our sample that differed from those reported previously. When the factors were converted to scales using unit scoring, the average value suggested a tendency toward a higher fat eating pattern. Results indicate that although behavioral domains appear to be constant across populations, fat-related eating patterns are not. These observations have implications for understanding the diversity of fat-related dietary patterns across groups and for planning appropriate behavior change strategies. PMID- 9216573 TI - What do consumers really think about dietary fat? AB - Nutrition communications in recent years have placed a priority on reducing dietary fat. Survey findings suggest this priority has fostered consumer obsession with and confusion about dietary fat and contributed to misperceptions about healthful eating. The obsession, confusion, and misperceptions about dietary fat and healthful eating, in turn, have created obstacles to achieving dietary goals. This overview of consumer knowledge, attitudes, and behaviors regarding dietary fat may allow dietetics and other professionals to tailor nutrition communication efforts toward clearing the confusion and better fostering success in reaching dietary goals. Nutrition communicators are encouraged to work together to restore reason to nutrition messages and recommendations and joy to food and eating in an effort to help consumers truly achieve nutrition and health goals. PMID- 9216574 TI - Can you have your low-fat cake and eat it too? The role of fat-modified products. AB - The American public, along with the medical and scientific communities, has certain expectations about the consumption of fat-modified foods; specifically, that such consumption will result in positive health benefits for both the individual and the population. Initial attempts by consumers to reduce fat intake required elimination of favorite foods or substitution of those foods with less palatable offerings. The food industry now has developed more than 5,600 reduced fat products of varying palatability. However, recent questions have arisen regarding the potential use and anticipated health benefits of these products. This commentary explores the underlying assumptions and expectations surrounding the use of fat-modified products, examines current usage rates of these products, and reviews the reported impact of these products on overall diet by relating these issues to two theoretic frameworks (Diffusion of Innovations and Stages of Change). Finally, some suggestions regarding realistic expectations for these products in the context of an overall diet are presented. PMID- 9216575 TI - The role of low-fat diets and fat substitutes in body weight management: what have we learned from clinical studies? AB - The introduction of low-fat, high-complex carbohydrate diets far the prevention and treatment of obesity was based on the causal link established between dietary fat and body fatness. Observational and mechanistic studies show that because fat possesses a lower satiating power than carbohydrate and protein, a diet rich in fat can increase energy intake. The propensity to gain weight is enhanced in susceptible persons, particularly sedentary people who have a genetic predisposition to obesity. Low-fat diets cause weight loss proportional to pretreatment body weight in a dose dependent manner; that is, weight loss is correlated positively to the reduction in dietary fat content. A reduction of 10% fat energy produces an average 5-kg weight loss in obese persons. As with traditional caloric counting diets, obese persons lose weight only if they adhere to the prescribed low-fat diet. Failure to achieve a weight loss and to maintain it may be attributed in part to lack of adherence to the diet. After a major weight loss, an ad libitum low-fat diet program appears to be superior to caloric counting in maintaining the weight loss 2 years later. Replacing some fat with protein instead of carbohydrate may increase the weight loss further. Moreover, fat substitutes may make it easier to prevent and treat obesity by making the diet palatable. More randomized, controlled, long-term dietary intervention studies are warranted to identify the optimal diet composition for the treatment of obesity. PMID- 9216576 TI - Low-fat and delicious: can we break the taste barrier? AB - No matter how healthy a food or recipe might be, if it does not taste good, people will not eat it. The professional and personal challenge is to find ways to cook favorite foods in a low-fat and delicious style to maintain health while enjoying foods without guilt. A number of strategies can be employed to reach that goal, including ingredient choices, low-fat product substitutes, cookware, cooking techniques, seasonings, basic cooking and recipe modifications, and the creation of new recipes. PMID- 9216577 TI - Reducing dietary fat: putting theory into practice--conference summary. PMID- 9216578 TI - Abdominal aortic aneurysms: who should we be screening? PMID- 9216579 TI - "Best practice" in surgical management of breast cancer. PMID- 9216580 TI - Organ donation: how can we improve the rates? PMID- 9216581 TI - Surgical management of breast cancer in Australian women in 1993: analysis of Medicare statistics. AB - OBJECTIVE: To examine patterns of surgical management of breast cancer among Australian women. DESIGN: Retrospective survey of Medicare records (a national dataset of all services rendered on a "fee-for-service" basis for which a Medicare benefit has been paid). PATIENTS: All Australian women (4683) who underwent surgery consistent with being for breast cancer in 1993 and for which Medicare benefits were paid. MAIN OUTCOME MEASURES: Proportions of women undergoing different forms of mastectomy, breast-conserving surgery and axillary surgery by patient age and State and region (urban or rural) of residence. RESULTS: Modified radical mastectomy was the most common surgery, performed in 2097 of the 4683 women (44.8%), while 1868 (39.9%) had breast-conserving surgery. Frequency of breast conservation decreased significantly with age and varied significantly between States and region of residence. It ranged from 34% in Western Australia to 49% in South Australia and the Northern Territory, and from 34% among rural women to 42% among urban women. Axillary surgery was recorded for 83% of all women studied. CONCLUSIONS: There was substantial geographical variation in patterns of surgical management for breast cancer. The tendency for rural women to undergo mastectomy rather than breast-conserving surgery may reflect the relative lack of access to postoperative radiotherapy. We are unable to explain the variation between States. PMID- 9216583 TI - Phaeochromocytoma with adult respiratory distress syndrome mimicking septicaemic shock. AB - Phaeochromocytomas are rare tumours that most commonly present with chronic sustained hypertension and hypertensive paroxysms or crises. We report a 49-year old woman with unsuspected phaeochromocytoma who presented with sudden onset of profound hypotension and adult respiratory distress syndrome unresponsive to fluid and inotropic support. This case illustrates the diversity of presentations of phaeochromocytoma, depending on the type and amount of catecholamines secreted. PMID- 9216582 TI - Prevalence of abdominal aortic aneurysms in men with diabetes. AB - OBJECTIVE: To assess the prevalence of abdominal aortic aneurysm (AAA) in men with diabetes aged 60 years and over. DESIGN: Prospective screening study. PATIENTS: 303 eligible participants among the first 1000 recruited to a large, community-based study of diabetes. MAIN OUTCOME MEASURES: Aortic diameter > or = 30 mm on screening ultrasonography, or previous surgery for AAA. RESULTS: AAA was diagnosed in three of the 303 men screened, and four others had previously had surgery for AAA. The aorta was not visualised in three obese men. Only one AAA required surgery (> or = 50 mm diameter). The overall prevalence of AAA was 2.3% (7/300), lower than that reported previously in the general population. Multivariate logistic regression analysis showed statistically significant associations with fasting triglyceride levels, and a history of intermittent claudication. CONCLUSIONS: Although a small number of men with diabetes aged 60 or more have undiagnosed AAA, the prevalence does not appear to be high enough to warrant targeted ultrasound screening. PMID- 9216584 TI - The methodology of cost-effectiveness analysis: avoiding common pitfalls. AB - Clinicians need to be familiar with the core methodology of cost-effectiveness economic analysis, as this information will increasingly be used in clinical decision-making. Just as in clinical trials, there are rules to be followed, and many pitfalls for the novice. PMID- 9216585 TI - Withdrawal of nifedipine capsules: jeopardizing the treatment of acute severe hypertension in pregnancy? Australasian Society for the Study of Hypertension in Pregnancy. AB - Short-acting oral nifedipine has been withdrawn from the Australian market because of reports of its adverse effects after long-term treatment in non pregnant patients with heart disease. This will have a major impact on the treatment of acutely hypertensive pregnant women, in whom the drug has proven to be safe, effective and easy to administer. Should pregnant women be forced to use less suitable agents, thus threatening their own and their babies' health? PMID- 9216587 TI - Making gains in health. PMID- 9216586 TI - Guidelines for managing patients with unstable angina. Rating the evidence and rationale for treatment. PMID- 9216588 TI - Clinical spirometry. PMID- 9216589 TI - Spontaneous bilateral Achilles tendon rupture associated with ciprofloxacin. PMID- 9216590 TI - Sexually transmitted diseases and unwanted pregnancies in chronically ill psychiatric patients. PMID- 9216591 TI - Sexually transmitted diseases and unwanted pregnancies in chronically ill psychiatric patients. PMID- 9216592 TI - Sexually transmitted diseases and unwanted pregnancies in chronically ill psychiatric patients. PMID- 9216593 TI - Prevalence of childhood sexual abuse in a community sample of Australian women. PMID- 9216594 TI - Women and men and the medical workforce in Australia. PMID- 9216595 TI - Safe drinking water. PMID- 9216596 TI - Why aren't all pregnant women being tested for HIV? PMID- 9216597 TI - Protecting eyes from sun damage. PMID- 9216598 TI - Identifying the active general practice workforce through public domain databases. PMID- 9216599 TI - The malaria frontline. Pioneering malaria research by the Australian Army in World War II. PMID- 9216600 TI - Pleuritic pain and pulmonary embolism in the emergency department: diagnostic and treatment issues. PMID- 9216601 TI - Social class and male cancer mortality in New Zealand, 1984-7. AB - Social class differences in cancer mortality among New Zealand men aged 15-64 years are examined for the period 1984-7. Age-standardised rates are presented for all cancer deaths, and for 23 specific cancer sites. The strongest social class mortality gradients were found for cancers of the larynx, liver, buccal cavity/pharynx, oesophagus, lung and for soft tissue sarcoma. On the other hand, rectal cancer, malignant melanoma, colon cancer, brain/nervous system cancers, and multiple myeloma showed higher death rates for the more advantaged socioeconomic groups. Lung cancer accounted for 54.1% of the overall social class gradient, and the major smoking related cancers (these include buccal/pharynx, oesophagus, larynx, lung and bladder, although it should be stressed that not all cases of these cancers are caused by smoking) accounted for 77.6% of the overall gradient. PMID- 9216602 TI - Renal transplantation in children; the Auckland experience 1980-96. AB - AIM: To review the experience and outcome of renal transplantation in children who received a renal allograft at the Auckland Children's Hospital between January 1980 and June 1996. METHODS: Retrospective review of patient records and access to data collected through the ANZDATA Registry. Forty one renal transplants were undertaken in 39 children. Prednisone and Azathioprine were the mainstay of the immunosuppressive regimen from 1980 to 1985, then cyclosporin A was introduced in 1986. Median age of first graft was 11 years (range 1.6-14 years). There were 34 living related and 7 cadaveric grafts and 17 males and 22 females. Thirteen children (33%) had a primary glomerular disorder as the cause of renal failure. Another 9 (28%) had reflux nephropathy and a further 8 (18%) had congenital renal dysplasia/hypoplasia either as primary cause or secondary to lower urinary obstruction. RESULTS: Survival analysis showed that 93% of grafts were functioning at 12 months and 73% were still functioning at 5 years. Longest surviving transplant is 16 years. Ten primary grafts were lost with the most common cause being chronic rejection, accounting for 50% of all transplants lost. Recurrence of primary disease was the second most common (4 of 13) cause of graft failure. There was one malignancy in this series with a child developing a Kaposi's sarcoma. CONCLUSIONS: Renal transplantation is a successful treatment of endstage renal failure in children. Chronic rejection remains a major cause of graft loss and better immunosuppressive strategies are required to deal with this problem. PMID- 9216603 TI - Developmental outcome at 18 months of children less than 1000 grams. AB - AIMS: Aims of this paper were to carry out an audit of 105 extremely low birth weight infants at 18 months of age, identifying problems, disseminating the resulting information and providing a basis for future work. METHODS: Children born in 1990-2 were classified in categories I to IV according to outcome, and selected perinatal variables were analysed for these groupings. RESULTS: The disability rate (categories I and II) within this cohort was 21% a similar finding to that reported in other literature. For the group with slow motor development and/or tonal abnormalities the percentage was 15. Despite the high risk nature of this group 64% of children were progressing well (category IV) at this age. No significant differences in outcome were found between small for gestational age and appropriate weight for gestational age infants. Significant results were demonstrated in the adverse effects of chronic lung disease and intraventricular haemorrhage on subsequent development. CONCLUSION: The information obtained from this study provides support for the use of this type of audit in Units with extremely low birth weight populations. PMID- 9216604 TI - How many neonates in New Zealand would qualify for extracorporeal membrane oxygenation? AB - AIM: To estimate how many neonates in New Zealand would qualify for extracorporeal membrane oxygenation (ECMO), using both standard published criteria and locally derived criteria. METHODS: Retrospective chart review of all babies with a birth weight over 2000 g admitted to neonatal intensive care in Auckland from June 1990 to June 1993. Ventilation and blood gas indices were calculated for all babies who were ventilated in 100% O2 for more than 4 hours and who met the basic criteria for ECMO (less than 1 week old with no neurological or chromosomal problems). These indices were compared with published ECMO criteria. Using a threshold of an 80% mortality, Auckland criteria for ECMO were derived. RESULTS: Of the published criteria for ECMO, only an oxygenation index of greater than 40 for 4 hours predicted a mortality of more than 80% in our population. From our own findings a PaO2 < 6.5 kPa for 4 hours predicted a mortality of 79%. CONCLUSION: Approximately 19 neonates might qualify for ECMO in New Zealand each year. PMID- 9216605 TI - Can preterm twins breast feed successfully? AB - AIM: To compare the success of singleton and twin preterm infants in establishing and maintaining breast feeding, and to evaluate the effectiveness of current programmes to promote breast feeding. METHODS: All infants less than 37 weeks gestation discharged in one month from the special care baby unit at National Womens Hospital were studied. Data on the infants and their in hospital course was recorded from the neonatal records. The mothers were contacted by telephone 3 to 4 months after discharge, to elicit the subsequent breast feeding rates. RESULTS: Thirty of 33 preterm infants (29 to 36 weeks gestation) were breast fed at discharge from hospital: 93% of singletons, and 89% of twins. The twins were older and heavier at discharge (p < 0.004) due to their longer hospital stays (28.4 vs 16.3 days, p < 0.05). All but 2 infants progressed to exclusive breast feeding. There was a similar rate of decline in the rates of breast feeding in singletons and twins to 68% at 8-12 weeks and 49% at 12-16 weeks after birth. CONCLUSIONS: Preterm twins can breast feed as successfully as preterm singleton infants; as with sufficient assistance and encouragement, their rates of breast feeding were comparable to those of term infants. Although the resources of this hospital do not allow preterm infants to become fully breast fed before discharge, the current programme at National Womens Hospital is effective in establishing successful breast feeding in these high risk infants. PMID- 9216606 TI - The effect of a preanaesthetic information booklet on patient understanding and satisfaction. AB - AIM: To determine if a brief user friendly anaesthetic booklet compliments the anaesthetic service currently provided, in terms of greater patient understanding and satisfaction. METHOD: Two questionnaires were completed by participants in each group, one questionnaire preoperatively and the other postoperatively. The booklet group received the anaesthetic booklet in the mail with their booking card while the control group only received their booking card. RESULTS: Of the 209 eligible, 140 patients consented to and completed the preoperative questionnaire, of whom 53 were in the anaesthetic booklet group and 87 were in the control group. The postoperative questionnaire was completed by 38 and 65 respectively. The anaesthetic booklet group had better understanding of what a premed will do (p < 0.05) and how long after an anaesthetic to wait before driving (p < 0.025). The percentage of correct answers for the more general anaesthetic questions was high and very similar in both groups. There was no significant difference in the satisfaction scores between groups. Satisfaction scores for both groups rose significantly in the postoperative questionnaire when compared with the preoperative questionnaire (p < 0.001). CONCLUSION: The value of the anaesthetic booklet is in providing detailed anaesthetic information to the patient. This will aid the preanaesthetic consultation with the anaesthetist and provide a focus for further discussion about the intended anaesthetic. Patient satisfaction with the anaesthetic service was high in both groups pre- and postoperatively and was not altered by the anaesthetic booklet. PMID- 9216607 TI - Wooden foreign bodies in the hand. PMID- 9216608 TI - Cannabis and violence. PMID- 9216609 TI - Manuka honey and leg ulcers. PMID- 9216610 TI - Aetiology of Parkinson's disease. PMID- 9216611 TI - Carbon dioxide accumulation in cots. PMID- 9216612 TI - The power line-cancer debate: is it a conflict between physics and biology? PMID- 9216613 TI - Can low-level 50/60 Hz electric and magnetic fields cause biological effects? AB - Some epidemiological studies have suggested that exposure to ambient, low-level 50/60 Hz electric and magnetic fields (EMFs) increases risk of disease. Whether this association has a causal basis depends in part on whether the electrical, chemical and mechanical "signals" induced within living cells by ambient EMFs are detectable in the complex milieu of voltages, currents and forces present within the living organism. Magnetic responsiveness has been found in some animals and bacteria; aquatic animals (e.g. sharks and rays) can sense weak electric fields. We outline the physics of several mechanisms by which EMFs may interact: (1) Energy transfer by acceleration of ions and charged proteins modifies cell membranes and receptor proteins; however, EMF energies are far below those typical of biomolecules in the cell. (2) Electric fields induced inside the body exert force on electric charges and electric moments; however, these forces are considerably smaller than typical biological forces. (3) The magnetic moments of ferromagnetic particles and free radical molecules interact with magnetic fields, but magnetic-moment sensory cells have not been found in humans, and modification of radical recombination rates by EMFs in a biological system is highly problematic. (4) Resonant interactions involve EMFs driving vibrational or orbital transitions in ion-biomolecule complexes; these mechanisms conflict with accepted physics, and many experimental tests have not found the predicted effects. (5) Temporal averaging or spatial summation can improve the ratio of "signal" to "noise" in any system, but this "mechanism" requires biological structures and neural processes having the necessary capabilities of EMF detection and temporal averaging that have not been found in humans. In summary, biological effects in humans due to extremely low-frequency EMFs of the order of those found in residential environments [< or = 2 microT (< or = 20 mG)] are implausible based on current understanding of physics and biology. Biological effects in humans at higher fields [> 10 microT (> 100 mG)] might reach plausibility as a result of time-averaging in combination with a magnetic-moment transduction mechanism; but even here, neither specialized EMF transduction structures nor appropriate averaging networks have been demonstrated. The bypothesis that the epidemiological associations observed between 50/60 Hz EMFs and disease reflect a causal relationship is not supported by what is known about mechanisms. PMID- 9216614 TI - Interaction of DNA-dependent protein kinase and poly(ADP-ribose) polymerase with radiation-induced DNA strand breaks. AB - Two of the enzymes involved in the response of mammalian cells to ionizing radiation are the DNA-dependent protein kinase and poly(ADP-ribose) polymerase. These enzymes are known to be activated by binding to DNA strand breaks, but previous studies designed to look at strand break specificity have employed enzymatically generated strand breaks and not irradiated DNA. Using highly purified DNA-dependent protein kinase, we compared enzyme activation by a series of DNA substrates. Irradiated plasmid DNA activated DNA-dependent protein kinase in a dose-dependent manner. When calculated in terms of the molar concentration of double-strand breaks, the enzyme activation by irradiated DNA was comparable to that by restriction enzyme-cleaved DNA. Linear DNA purified after plasmid irradiation also activated DNA-dependent protein kinase to a comparable extent, but nicked DNA, either isolated from irradiated plasmid or generated by DNase I, failed to activate the enzyme. A comparison of the enzyme activation by plasmid molecules with different 3'- and 5'-terminal groups indicated that the chemical nature of the DNA termini at the double-strand break does not significantly influence the response of the DNA-dependent protein kinase. Similar experiments with poly(ADP-ribose) polymerase demonstrated that single- and double-strand breaks activate this enzyme with almost equal efficiency, but because of their greater number, single-strand breaks dominate the response of poly(ADP-ribose) polymerase to irradiated DNA. PMID- 9216615 TI - Radiation-induced DNA damage in tumors and normal tissues: IV. Influence of proliferation status and cell type on the formation of oxygen-dependent DNA damage in cultured cells. AB - Using a variety of techniques, several laboratories have recently demonstrated the feasibility of using radiation-induced DNA strand breaks and/or DNA-protein crosslinks (DPCs) to detect and/or quantify hypoxic cells in tumors and normal tissues. However, if strand breaks and/or DPCs are to be used to estimate the hypoxic fraction or the fractional hypoxic volume of tumors and normal tissues, their formation as a function of the oxygen concentration near the DNA must be relatively independent of the biological properties of these cells. In the present study, the shape of the oxygen dependence curves and the K(m) values for radiation-induced strand breaks and DPCs were measured by alkaline elution for proliferative (P) and quiescent (Q) cells of the mouse mammary adenocarcinoma, line 66. The sigmoidal shape of the oxygen dependence curves, the K(m) for strand breaks (approximately 0.027 mM) and the K(m) for the formation of DPCs (approximately 0.020 mM) were identical for the P and Q cells of line 66. Consequently, the proliferative status of these tumor cells had no measurable influence on the oxygen-dependent formation of radiation-induced strand breaks and DPCs. In addition, the percentage of the DNA retained on the filters after approximately 24 ml of elution without proteinase K in the lysis solution, a parameter equal to the sum of the strand breaks and DPCs that has been shown to be proportional to the percentage of hypoxic cells in the sample, was not significantly different for fully oxygenated or fully hypoxic populations from five tumor cell lines that varied in species, site of origin, proliferative status and/or properties of the proteins which are intimately associated with their DNA. These data indicate that the formation of radiation-induced strand breaks and DPCs depends predominantly on the oxygen concentration in the microenvironment around the DNA, and only minimally on the biological properties of the cells. PMID- 9216616 TI - Longevity of pimonidazole adducts in spontaneous canine tumors as an estimate of hypoxic cell lifetime. AB - The longevity of pimonidazole adducts in tumors was quantified as an estimate of the lifetime of hypoxic cells. Pimonidazole was given before irradiation to 12 dogs bearing spontaneous tumors, and tumors were biopsied 24, 48 and 72 h later. Pimonidazole antigen was quantified in the biopsies using ELISA and immunohistochemistry. Pimonidazole antigen was detectable in the initial biopsy in all dogs. In 5 dogs the amount of detectable antigen decreased to less than 50% of the initial amount, in 5 other dogs the amount of detectable antigen decreased to an amount between 50 and 100% of the initial amount, and in 2 dogs the amount of antigen appeared to increase relative to the initial amount. Tumors with high initial adduct concentration were characterized by greater decreases in adduct concentration than tumors with low initial adduct concentration. Immunohistochemically, labeled cells were present in 11 of 12 tumors. The geographic area in tumor biopsies labeled immunohistochemically with pimonidazole adducts (labeled area fraction) tended to decrease over time in 6 dogs, remain stable in 4 dogs and seemingly increase in 1 dog. There was no relationship in individual tumors between the relative change in antigen concentration and the relative change in labeled area fraction. Hypoxic cells which bind pimonidazole may persist for days during fractionated radiation therapy, and the potential exists for them to exert a negative effect on the host. PMID- 9216617 TI - Sonodynamic toxicity of gallium-porphyrin analogue ATX-70 in human leukemia cells. AB - Low concentrations (> or = 1 microM) of the gallium-porphyrin analogue ATX-70 significantly enhanced cellular toxicity in human leukemia HL-525 cells exposed to 50 kHz ultrasound. The mechanism of this ATX-70-dependent sonosensitization is unknown, but we have established the requirement of extracellular localization of ATX-70 molecules for sonosensitization. Short-lived toxic intermediates produced from ATX-70 by ultrasound are implicated in the mechanism, since no cytotoxicity was found when medium containing ATX-70 was sonicated and subsequently added to the cells. However, we were unable to demonstrate the existence of radical intermediates by EPR spin trapping with the nitroso spin trap, DBNBS, and ATX-70 dependent sonotoxicity could not be ameliorated by the addition of up to 70 mM POBN and DMPO spin traps during ultrasound exposure. PMID- 9216618 TI - Schedule dependence of the interaction of radiation and Taxol in HeLa cells. AB - Previous studies have indicated that the nature of the interaction of radiation and Taxol may be dependent on the cell line, drug dose and treatment schedule. The present study represents a systematic attempt to examine the schedule dependence of the interaction of radiation and Taxol in HeLa cells. The protocol used was radiation treatment (7 Gy), followed by a variable interval (0-24 h), followed by exposure to Taxol (7.5 nM, 24 h), followed by plating for a colony forming assay. Parallel samples were also taken for flow cytometric analysis of the distribution of cells in the phases of the cell cycle at the beginning and end of Taxol treatment. The results indicate that sub-additive, additive or supra additive interaction can be seen depending on the interval between the radiation and Taxol treatments. Full radiation survival curves were determined for cells exposed to Taxol either immediately or 10 h after the completion of radiation treatment, corresponding to sub-additive and supra-additive interactions, respectively. An examination of the data revealed that maximum cell killing occurred when the percentage of cells in G1 phase was at a minimum at the time of addition of Taxol. Studies of Taxol-induced toxicity using cells synchronized in G1 phase with mimosine and then released and allowed to progress through the cell cycle confirmed this observation. The conclusion of this study is that prior radiation treatment can modify the effect of subsequent Taxol treatment through alterations in the distribution of cells in the phases of the cell cycle. This finding has important implications for the clinical scheduling of these two cancer treatment modalities. PMID- 9216620 TI - A mortality study of employees of the nuclear industry in Oak Ridge, Tennessee. AB - An analysis was conducted of 27,982 deaths among 106,020 persons employed at four Federal nuclear plants in Oak Ridge, Tennessee, between 1943 and 1985. The main objectives were to extend the evaluation of the health effects of employment in the nuclear industry in Oak Ridge to include most workers who were omitted from earlier studies, to compare the mortality experience of workers among the facilities, to address methodological problems that occur when individuals employed at more than one facility are included in the analysis, and to conduct dose-response analyses for those individuals with potential exposure to external radiation. All-cause mortality and all-cancer mortality were in close agreement with national rates. The only notable excesses occurred for white males for lung cancer [standardized mortality ratio (SMR) = 1.18, 1,849 deaths] and non malignant respiratory disease (SMR = 1.12, 1,568 deaths). A more detailed analysis revealed substantial differences in death rates among workers at the Oak Ridge plants. Evaluation of internally adjusted log SMRs using Poisson regression showed that workers employed only at Tennessee Eastman Corporation or K-25 and at multiple facilities had higher death rates than similar workers employed only at X-10 or Y-12, and that the differences were primarily due to non-cancer causes. Analysis of selected cancer causes for white males indicated large differences among the workers at the different facilities for lung cancer, leukemia and other lymphatic cancer. Dose-response analyses for external penetrating radiation were limited to a subcohort of 28,347 white males employed at X-10 or Y-12. Their collective recorded dose equivalent was 376 Sv. There was a strong "healthy worker effect" in this subcohort-all-cause SMR = 0.80 (4,786 deaths) and all cancer SMR = 0.87 (1,134 deaths). Variables included in the analyses were age, birth cohort, a measure of socioeconomic status, length of employment, internal radiation exposure potential and facility. For external radiation dose with a 10 year lag, the excess relative risk was 0.31 per Sv (95% CI = -0.16, 1.01) for all causes and 1.45 per Sv (95% CI = 0.15, 3.48) for all cancer. The estimated excess relative risk for leukemia was negative but imprecisely determined. A preliminary dose adjustment procedure was developed to compensate for missing dose but not other dosimetry errors. Results of the analyses using the adjusted doses suggest that the effect of missing dose is an upward bias in dose-response coefficients and test statistics. PMID- 9216619 TI - Issues in the comparison of risk estimates for the population in the Techa River region and atomic bomb survivors. AB - Plutonium production in the former Soviet Union began in 1949 at the Mayak Production Association located between the cities of Chelyabinsk and Ekaterinbourg in the southern Ural mountains about 1200 km east to Moscow. During the first few years of Mayak's operation, almost 30,000 people living on the banks of the Techa River received significant internal and external exposures as a consequence of the release of large quantities of radioactive materials from Mayak. Studies of levels of radioactive contamination and health effects in this population began in the early 1950s. A systematic follow-up of a fixed cohort that includes all people who were living in Techa River villages in 1949 was begun about 30 years ago. In this paper we describe the Techa River cohort, outline the nature of the exposures and discuss the status of follow-up for the period from 1950 through 1989. While noting the limitations of the current epidemiological follow-up data, we also compare the demographic and mortality structure of the Techa River cohort with the Life Span Study cohort of Japanese atomic bomb survivors. It is seen that, despite a number of limitations, the current data suggest that the risks of mortality from leukemia and other cancers increase with increasing radiation dose in the Techa River cohort. This finding suggests that, with continued improvements in the quality of the follow-up and dosimetry, the Techa River cohort has the potential to provide quantitative estimates of the risks of chronic low-dose-rate radiation exposures for an unselected general population that will be an important complement to the estimates based on the Life Span Study that are used as the primary basis for numerical assessments of radiation risk. PMID- 9216621 TI - Rb and p53 gene deletions in lung adenocarcinomas from irradiated and control mice. AB - This study was conducted on mouse lung adenocarcinoma tissues that were treated with formalin and embedded in paraffin 25 years ago to investigate the large gene deletions of Rb and p53 in B6CF1 male mice. A total of 80 lung tissue samples from irradiated mice and 40 lung samples from nonirradiated controls were selected randomly and examined in the Rb portion of this study. The results showed a significantly (P < 0.05) higher percentage of Rb deletions in lung adenocarcinomas from mice exposed to 60 once-weekly gamma-ray doses than those from mice receiving 24 once-weekly gamma-ray doses at low doses and low dose rates; however, the percentage was not significantly different (P > 0.05) from that for spontaneous lung adenocarcinomas or lung adenocarcinomas from mice exposed to single-dose gamma irradiation at a similar total dose. Rb fragments 3 (71%) and 5 (67%), the parts of the gene that encoded the pocket binding region of Rb protein to adenovirus E1A and SV40 T-antigen, were the most frequently deleted fragments. Analysis of p53 gene deletion was carried out on normal lungs and lung adenocarcinomas that were initially found to bear Rb deletions. Exons 1, 4, 5, 6 and 9 were chosen to be analyzed. The data showed that 30 (97%) of 31 normal lungs and lung adenocarcinomas had p53 deletions. Exons 4 (83%) and 5 (90%) were the most frequently deleted among tested exons. Mice exposed to neutrons 60 times on a once-weekly schedule had a higher percentage of complete p53 deletions (5/8; 63%) than those exposed to gamma rays 60 times on a once weekly schedule (2/8; 25%). We conclude that p53 deletions may be one of the major mutational events in the tumorigenesis of lung adenocarcinomas in the irradiated B6CI, mice. PMID- 9216622 TI - Comparative clastogenic sensitivity of respiratory tract cells to gamma rays. AB - To understand the relationships between exposure and damage to different cell populations in the respiratory tract, methods were developed to culture deep-lung fibroblasts and epithelial cells from the nose, trachea and deep lungs. Female F 344 Fischer and male Wistar rats were exposed to 1-5 Gy of 60Co gamma rays at a dose rate of 0.4 Gy/min. Cells were isolated for short-term culture, and the incidences of binucleated cells and micronuclei were determined. The incidences of micronuclei were determined in cytochalasin-B-induced binucleated cells at 72 h for nasal and tracheal tissue and 96 h for deep-lung fibroblasts and epithelial cells. Maximum frequencies of binucleated cells were found in the control nonirradiated cells at these harvest times, and the frequencies were not significantly affected at these harvest times by radiation exposure. No significant differences were found in the frequencies of micronuclei induced in the nasal epithelial cells isolated from female F-344 Fischer or male Wistar rats. Fibroblasts cultured in different media and isolated from either female F 344 Fischer or male Wistar rats also showed a similar frequency of micronuclei. Over the doses tested, the frequency of micronuclei in the respiratory tract cells increased linearly with the dose. The slopes were 92.2 +/- 9.2, 76.2 +/- 7.9, 32.8 +/- 2.4 and 28.7 +/- 3.4 micronuclei/1000 binucleated cells/Gy for deep lung epithelial cells, deep-lung fibroblasts, tracheal epithelial cells and nasal epithelial cells, respectively. Deep-lung epithelial or fibroblast cells were about two to three times as sensitive for elastogenic damage as nasal and tracheal epithelial cells. The measurement of micronuclei in isolated respiratory tract cells is very useful in assessing cytogenetic damage induced in different cell types by radiation. PMID- 9216623 TI - Relationship between turnover of cesium-137 and dietary potassium content in potassium-restricted mice. AB - The biological half-life of 137Cs and its organ distribution were investigated in mice fed various potassium-deficient diets. The biological half-life, which was 6.1 days in mice receiving the normal level of potassium, became longer as the dietary potassium content decreased, and 137Cs was hardly excreted from the body when dietary potassium content was restricted to 200 mg/kg or less. The muscle showed the highest concentration of 137Cs in both mice that had sufficient amounts of potassium and those that were potassium-deficient. Clearance of 137Cs from tissues was generally suppressed when mice were fed a potassium-deficient diet, but the relative distribution pattern of 137Cs was not affected by dietary potassium content. These results suggest that dietary potassium intake, which may vary with eating habits, affects the biological half-life of 137Cs in humans. PMID- 9216624 TI - Sedation of children for emergency imaging. AB - Successful and safe sedation is an important technical aspect of pediatric imaging for the radiologist. Sedation of children requiring acute diagnostic evaluation presents additional challenges including uncertain past medical history and nothing-by-mouth status, unknown allergies, and unstable clinical status. This article focuses on the techniques and problems of sedating children in an acute setting. PMID- 9216626 TI - Pancreatic emergencies. AB - Sonography and CT scan remain the examinations of choice for evaluating pancreatic diseases. This article addresses the clinical and imaging features of the two most important pancreatic emergencies: (1) pancreatitis and (2) trauma. The rationale for selecting sonography or CT scan is also addressed. PMID- 9216625 TI - Acute hepatic disease. AB - Acute hepatobiliary dysfunction in children may occur as the result of trauma, infectious hepatitis, biliary obstruction, benign and malignant tumors, and acute vascular obstruction. The targeted and integrated use of modern imaging techniques has made a significant impact in the diagnosis and management of these disorders. PMID- 9216627 TI - Splenic emergencies. AB - Of medical and surgical emergencies in the pediatric abdomen, those involving the spleen are relatively less common than other abdominal organs, though equally important to recognize. A more sophisticated clinical understanding of the important role of the spleen in immunocompetence has developed in parallel with advancements in imaging. A healthy respect for the preservation of splenic tissue has emerged, altering traditional surgical management of splenic emergencies. Non invasive imaging has come to play a vital role in depicting acute abnormalities and in determining the need for conservative, interventional, or surgical management. PMID- 9216628 TI - Neonatal gastrointestinal emergencies. AB - Intestinal obstruction is the most common and important gastrointestinal emergency in the newborn period. This article presents a general approach to neonatal obstruction and discusses the most common specific cases of obstruction. The importance of the plain abdominal radiograph is emphasized. PMID- 9216629 TI - Gastrointestinal emergencies in older infants and children. AB - Gastrointestinal tract emergencies in older infants include traumatic and nontraumatic conditions. The imaging evaluation of these conditions strongly affects diagnosis and management. This article provides a clinical overview and reviews the rationale for imaging and important imaging features of these gastrointestinal tract emergencies. PMID- 9216630 TI - Acute gastrointestinal bleeding. AB - Diagnosis and intervention in pediatric GI bleeding is the shared responsibility of pediatric endoscopists, radiologists, and surgeons. Brisk hemorrhage, though alarming, is most often self-limited; few cases require urgent surgery before diagnostic evaluation is accomplished. The choice between endoscopic and radiographic evaluation varies with the differential diagnoses being considered and with local referral patterns. Many imaging options exist for assessing GI bleeding in children, but these options are generally narrowed by clinical history and age-appropriate differential possibilities. PMID- 9216631 TI - Uroradiologic emergencies in infants and children. AB - Selected topics are discussed that represent common reasons for performing emergency uroradiologic examinations in infants and children, including urinary tract infection, hematuria, urinary retention, intermittent ureteropelvic junction obstruction, spontaneous perforation of the augmented urinary bladder, and urethral trauma. Common complications of voiding cystourethrography in children are also discussed. PMID- 9216632 TI - Emergency obstetric and gynecologic ultrasound. AB - In conclusion, sonography plays a central role in imaging the obstetric and gynecologic emergencies of babies, girls, and adolescents. Transabdominal and transvaginal sonography each have their respective and combined strengths, which depend on the age, size, anatomy, and social and clinical situation of the patient to be imaged. With recent advances in sonographic technology, especially in the current use of higher-frequency transducers and the promise of three dimensional imaging, we are undoubtedly seeing more anatomy and pathology, and seeing it more clearly. In the sexually mature patient, transvaginal sonography provides an exceptional view of the normal uterus and adnexae and the myriad presentations of pelvic pathology. This article has reviewed the sonographic techniques that can be used in imaging the pediatric and adolescent pelvis, and has emphasized some of the many pathologic conditions that can be elucidated by pelvic sonography. PMID- 9216633 TI - The acute scrotum. AB - In the pediatric age group, color Doppler sonography appears to be as accurate as scintigraphy in evaluating causes of acute scrotal pain, provided that sensitivity for detecting low-velocity flow is adequate. Sonography has the advantage of providing anatomic information and it lacks radiation. Scintigraphy remains a reliable method of evaluating acute scrotal pain and should be used when color Doppler sensitivity for low-velocity, low-volume testicular blood flow is inadequate leading to doubt about the sonographic diagnosis. It is also advocated when examiner expertise with color Doppler sonography is limited. Regardless of which imaging study one prefers, it needs to be recognized that imaging of any type is not warranted in patients with a high clinical suspicion of torsion, because it delays immediate surgical treatment needed to prevent permanent damage. PMID- 9216634 TI - Interventional radiology in the acute pediatric abdomen. AB - This article addresses the most common types of interventional procedures performed in the pediatric abdomen. Nonvascular interventions are stressed because they are more common than vascular interventions. PMID- 9216635 TI - Transforming growth factor-beta receptors in human cancer cell lines: analysis of transcript, protein and proliferation. AB - Transforming growth factor-beta (TGF-beta) is a growth regulator which affects multiple cellular functions through the TGF-beta type I and type II receptor (TGF beta RI and TGF-beta RII) serine/threonine kinases. In this study, the expression of TGF-beta RI and RII was investigated by northern blot, reverse transcription polymerase chain reaction (RT-PCR) and western blot analyses in human breast (MCF 7 and T-47D), ovary (CP70 and OVT2) and prostate (LNCaP, DU145 and PC3) cancer cell lines. Northern blot analysis showed expression of TGF-beta RI and TGF-beta RII mRNAs in all cell lines examined except for MCF-7 and LNCaP, respectively. In contrast, RT-PCR showed that all of the cell lines examined express TGF-beta RI and RII transcripts using specific primer oligonucleotides. Western blot analysis demonstrated that all of the cancer cell lines examined express TGF-beta RI and RII proteins. Southern blot analysis showed there were differences in the restriction enzyme digestion patterns for TGF-beta RI and RII of T47D compared to MCF-7, PC3 and LNCaP. Also, the addition of TGF-beta 1 to the breast and prostate cancer cells inhibited colony formation in soft agarose. Our studies show that in cancer cells, relatively low levels of TGF-beta receptor transcripts may result in protein production and inhibition of cell proliferation by TGF-beta. PMID- 9216636 TI - Cellular targets of the anti-breast cancer agent Z-1,1-dichloro-2,3 diphenylcyclopropane: type II estrogen binding sites and tubulin. AB - Z-1,1-dichloro-2,3-diphenylcyclopropane (a.k.a. Analog II, AII) is a known anti breast cancer agent with apparent antiestrogenic effects and remarkably low toxicity in rodents. We have recently shown that AII and its major metabolite Z alpha-chlorochalcone (ZCC) inhibit proliferation of both estrogen-responsive and nonresponsive human breast cancer cells, suggesting its mechanism is not mediated by the type I estrogen receptor (ER). The present studies were performed to begin to define the molecular targets of AII and ZCC. Based on the compounds' structures and actions, we hypothesized that their effects could be due to interaction at type II estrogen binding sites (EBSII) and/or cellular microtubules. The affinities of AII ZCC and the positive control diethylstilbestrol (DES) for the ER (in MCF-7 and MCF-7/LY2 cells) and EBSII (in MCF-7, MCF 7/LY2, and MDA-MB231 cells) were determined with a whole cell assay for displacement of [3H]estradiol. The kinetics of their effects on cellular microtubules and cell cycle distribution of human breast cancer cells were measured by indirect immunofluorescence and flow cytometry. Their abilities to inhibit assembly of isolated tubulin in vitro were determined. AII, ZCC, and DES had similar affinities for the EBSII in the three cell lines. Neither AII nor ZCC displaced [3H]estradiol from the ER in MCF-7 cells, whereas DES did. The microtubule network of MDA-MB231 cells exposed to 100 microM AII or 10 microM ZCC began to disassemble within 1 hour of treatment and was completely diffuse after 6 hour of exposure to either drug. AII inhibited in vitro assembly of tubulin, with an IC50 of 6.7 +/- 0.9 microM, while ZCC was inactive below 40 microM. Both drugs caused accumulation of the cells in the G2/M phase of the cell cycle. The evidence suggests that the antitumor action of AII is mediated, at least in part, through the EBSII and/or perturbation of tubulin-microtubule dynamics. AII thus represents a new lead compound for design and discovery of novel antitumor agents directed against the EBSII and/or microtubules. PMID- 9216637 TI - Expression of CD44 standard and variant isoforms v5, v6 and v7 in human ovarian cancer cell lines. AB - The expression of standard CD44 protein (CD44std) and its splice variants v5, v6 and v7 was investigated in 43 human ovarian carcinoma cell lines by flow cytometry and immunocytochemistry using monoclonal antibodies raised against extracellular epitopes. Twenty six (60%) cell lines expressed CD44 std. Variant isoforms of CD44 were expressed in 12 of the 26 CD44 positive cell lines. All 12 cell lines expressed CD44 v5. In addition 6 cell lines expressed CD44 v6 and one of these expressed CD44 v7 simultaneously. No significant differences of CD44 expression were found between cell lines derived from solid tumors or ascites. In ovarian cancer cells splicing of CD44 v5 appears to be a prerequisite for expression of downstream variable exons. New acquisition of variable CD44 exons may be implicated in the tumorigenesis of ovarian cancer. The CD44 gene provides a biological model to study the role of alternative splicing in gynecologic malignancy. PMID- 9216638 TI - Effects of oophorectomy on interstitial fluid pressure, blood flow and vascularity in a rat mammary tumour. AB - Interstitial fluid pressure (IFP), tumour size, blood flow and vascularity were measured in dimethyl-benz-alpha-anthracene-induced rat mammary tumours after oophorectomy. IFP was measured in the tumours by the wick-in-needle technique, blood flow by the labelled microsphere technique and vascularity by fluorescence vascular staining. Tumour volume was determined by measuring two diameters. Oophorectomy resulted in a decrease in tumour volume preceded by a decrease in IFP, while no significant change was observed in total tumour blood flow. The vascular cross-sectional area seemed to decrease. Cell proliferation may be of importance for the high interstitial fluid pressure in this hormone dependent tumour. A reduced metabolic requirement in the regressing tissue, down-regulating the vascular capacity, may camouflage the influence of the reduced tissue pressure on blood perfusion. PMID- 9216639 TI - Effects of cytogenin, a novel microbial product, on embryonic and tumor cell induced angiogenic responses in vivo. AB - Cytogenin (8-hydroxy-3-hydroxymethyl-6-methoxyisocoumarin) is a new microbial product with antitumor and antirheumatoid arthritis effects in vivo when administered orally, although its mechanism(s) of action is not known well. Both neoplasia and rheumatoid arthritis are referred to as angiogenesis-dependent diseases. The aim of the present study was to investigate the effects of cytogenin on both physiological and pathological angiogenesis, using the growing chick embryo chorioallantoic membrane and mouse dorsal air sac assay systems, respectively. The microbial product at doses up to 100 micrograms/egg did not significantly affect embryonic angiogenesis when topically placed on the surface of the chorioallantoic membrane, suggesting that it has no effect on the physiological (or normal) angiogenic response. By contrast, systemic administration of cytogenin (100 mg/kg p.o., for 5 consecutive days) significantly suppressed angiogenesis induced by malignant tumor cells (S-180), one of pathological neovascularization, in a mouse dorsal air sac assay system. Pharmacokinetic studies in mice revealed that the maximal concentration of cytogenin in plasma after a single 100 mg/kg oral dose of the compound was 32 microM. In vitro experiments involving cultured vascular endothelial cells showed that cytogenin at concentrations determined by pharmacokinetic study, had little effect on plasminogen activator secretion, tube formation and the proliferation of endothelial cells. These results suggest that cytogenin is a novel oral antiangiogenic agent, that the mechanism of its antiangiogenic action contributes to its suppressive effects on both tumor growth and rheumatoid arthritis that we previously found, and that it could be developed as a potential therapeutic agent for cancer, rheumatoid arthritis and other angiogenesis-dependent disorders such as diabetic retinopathy. PMID- 9216640 TI - Synergistic effects of adriamycin and TALL-104 cell therapy against a human gastric carcinoma in vivo. AB - The single and combined antitumor effects of adriamycin and a major histocompatibility complex non-restricted human cytotoxic T cell line (TALL-104) were evaluated in severe combined immunodeficient (SCID) mice engrafted subcutaneously with a biopsy sample from a patient with a highly metastatic gastric carcinoma. Chemotherapy was initiated 3 weeks after tumor implantation, when local growth was advanced, and consisted of 2 weekly injections of adriamycin. gamma-irradiated (40 Gy) TALL-104 cells were administered daily for 2 weeks, starting 1 day after the end of chemotherapy. While TALL-104 cells or adriamycin alone did not inhibit tumor growth, synergistic antitumor effects were seen with the two treatments combined. These findings suggest that chemotherapy in conjunction with cell therapy are effective in overcoming tumor resistance to single therapeutic agents through mechanisms independent from the host immune system. PMID- 9216641 TI - Immunomodulating efficacy of combined administration of galactoside-specific lectin standardized mistletoe extract and sodium selenite in BALB/c-mice. AB - The immunomodulating abilities of commercially available mistletoe extract standardized for the galactoside-specific lectin (mistletoe lectin-1, ML-1) and sodium selenite (Se) were evaluated in BALB/c-mice and compared to non-treated control animals. Following the optimal schedule of administration (ML-1: 1ng/kg BW; days 1, 2, 3, 5 and Se: 3,5 micrograms/kg BW for 7 subsequent days before evaluation) yielded enhanced counts (thymocytes; peritoneal macrophages, MO; peripheral blood leukocytes; lymphocytes, PBL; monocytes, PBM) and activities (CD 25 positive PBL, MAC-3 positive PBM) of desired immune cells reaching statistical significance for most parameters. However, combined administration of ML-1 and Se proved to be superior to monotherapy since immune cell counts (thymocytes, leukocytes, PBL, PBM) and activities (CD-25 positive PBL) exceeded values obtained after monotherapy. These data are in favour of therapeutical strategies in complementary oncology and suggest that the combination of defined immunomodulating substances might positively enhance the efficacy. PMID- 9216642 TI - Optimal duration of whole body hyperthermia when combined with cis-diaminne-1,1 cychlobutane dicarboxylate platinum (II) (carboplatin). AB - Minimizing normal tissue toxicity can enhance the therapeutic gain of thermochemotherapy. For this purpose, we investigated the optimal duration of whole body hyperthermia (WBH) (41.5 degrees C) when administered simultaneously with carboplatin (CBDCA). Using a transplantable fibrosarcoma in Fischer 344 rats, we measured tumor growth delay (TGD) as well as normal tissue toxicities (body weight loss, thrombocytopenia) induced by various durations of WBH (0.5, 1.0, 1.5, 2.0 or 2.5 hours) when combined with CBDCA (30 mg/kg, i.v.). When combined with CBDCA, 1.0 hour WBH increased the TGD compared to 0.5 hour of WBH, but with WBH durations greater than 1.0 hour, the TGD did not further significantly increase. Measuring CBDCA-induced myelosuppression, the platelet count on day 6 post-treatment decreased from a control mean of 6.8 x 10(8)/ml to 1.8 x 10(8)/ml after 2.5 hour WBH exposure in a duration-dependent manner (p < 0.001). To estimate the specific therapeutic efficacy (STE), we calculated a ratio of TGD to myelosuppression (thrombocytopenia). Compared to other WBH exposure times, 1.0 hour duration of WBH combined with CBDCA produced the highest STE (2.8) and over 1.5 hour duration of WBH did not result in any additional increase in STE. We conclude that 1.0 hour WBH exposure is optimal when combined with CBDCA in order to maximize the therapeutic gain. PMID- 9216643 TI - Gliomas are driven by glycolysis: putative roles of hexokinase, oxidative phosphorylation and mitochondrial ultrastructure. AB - To elucidate the reasons for glycolytic deviation commonly found in brain tumors, hexokinase (HK) activity, mitochondria-HK binding, oxidative phosphorylation and mitochondrial ultrastructure were studied in 4 human xenografted gliomas. Lactate/pyruvate ratios were increased 3-4 fold and HK activity was of 2-4 fold lower than that of normal rat brain tissue, used as the control. The mitochondria bound HK (mHK) fraction varied considerably and represented 9 to 69% of the total HK of that normal rat brain. The respiratory activity of glioma mitochondria, assessed by polarography and spectrophotometry, was within the normal range. However, the mitochondrial content of gliomas was lower than in the rat brain tissue, as revealed by the markedly decreased, activities of two unrelated mitochondrial enzymes, cytochrome c oxidase and citrate synthase in glioma homogenates. Electron microscopical studies confirmed the reduced number of mitochondria in 3 out of the 4 gliomas. Profound alterations of mitochondrial ultrastructure, namely of cristae and matrix densities, were observed in the 4 gliomas. The intercrista space was wider in all gliomas and the crista area was larger in 3 out of the 4 gliomas than in normal rat brain. Finally, the outer membrane of glioma mitochondria interacted intimately and extensively with the rough endoplasmic reticulum (RER) and/or nuclear membrane. These results suggest that, because of the very low content of normally functioning mitochondria, gliomas shift their energy metabolism towards a high-level glycolysis to generate their cellular ATP supply, probably through RER-mitochondria interactions and transformation-dependent redistribution of particulate HK from non-mitochondrial to mitochondrial receptors. PMID- 9216644 TI - Chemosensitizing activity of caffeic acid in multidrug-resistant MCF-7/Dox human breast carcinoma cells. AB - The chemosensitizing activity of caffeic acid was examined in parent MCF-7 and multidrug-resistant MCF-7/Dox human breast carcinoma cells. In clonogenic assays, MCF-7/Dox cell was about 135-fold less sensitive to doxorubicin than MCF-7 cells. Caffeic acid (10 microM) slightly altered the colony-forming ability of MCF-7 cells, and markedly reduced the IC50 of doxorubicin (Dox) from 10.8 +/- 1.3 microM to 0.83 +/- 0.21 microM in MCF-7/Dox cells. When compared to MCF-7/Dox cells, intracellular accumulations of [14C] Dox in MCF-7 cells for 1 hour and 12 hours were elevated 2.3-fold and about 6.4-fold, respectively. Doxorubicin accumulations in MCF-7 and MCF-7/Dox cells were not altered in the presence of 10 microM caffeic acid. Both TGF beta 1 and TGF beta 2 isotypes were detected in MCF 7/Dox cells, while only TGF beta 1 was found in MCF-7 cells. The level of TGF beta 1 in MCF-7/Dox cells was about 3-fold greater than that in MCF-7 cells. In cells pretreated with caffeic acid (10 microM), TGF beta 1 and TGF beta 2 levels were overexpressed only in MCF-7/Dox cells by 90% and 60%, respectively. These results suggest that caffeic acid is potentially a chemosensitizing agent with greater selectivity to drug-resistant MCF-7/Dox cells over parent MCF-7 cells and that the chemosensitizing effect is not mediated by altered drug concentrations in the cells, but may be possibly correlated to the induction of TGF beta isotypes. PMID- 9216645 TI - Mechanism of growth inhibition by calpain inhibitor in MCF-7 cells. AB - BACKGROUND: A calpain inhibitor, calpeptin, inhibited the cell growth of ER (estrogen receptor) positive breast cancer cells, such as MCF-7, T-47D, and ZR-75 1 in the presence of E2. The mechanism of this inhibition has not been clarified yet. MATERIALS AND METHODS: MCF-7 cells were employed to investigate the mechanism of the inhibition. We studied the effect of calpeptin on the secretion of insulin-like growth factor-I (IGF-I) and transforming growth factor (TGF alpha). RESULTS: The secretion of IGF-I or TGF-alpha was not changed by calpeptin either in the presence or absence of E2. Moreover, the binding of IGF-I or TGF alpha to MCF-7 cells augmented by the addition of E2 was not affected by calpeptin. CONCLUSIONS: These results indicated that the inhibition of cell growth in MCF-7 by calpeptin was not due to the modulation of autocrine growth factors and their receptors. PMID- 9216646 TI - The epidermal growth factor receptor is involved in autocrine growth of human bladder carcinoma cell lines. AB - We have previously shown that human bladder tumor cell lines may be adapted to grow in the complete absence of serum or any other growth supplement and that this can be explained on the basis of autocrine stimulation. In the present study we have extended the number of cell lines that could be established as serum-free cultures and found this capacity to be correlated with tumor malignancy. We also used the receptor blocking monoclonal antibody, mAb 528, to study its effect on tumor cell growth. Inhibition was observed in all of seven bladder carcinoma cell lines tested. A similar effect was observed in two colon carcinoma cell lines but not in a melanoma line. The results show that the EGFR is involved in autocrine growth stimulation and that the acquirement of autonomous growth capacity is likely to be an important factor in the oncogenesis of bladder tumors. PMID- 9216647 TI - Transformation of human glioma cell lines with the p16 gene inhibits cell proliferation. AB - We examined the genomic status of the p16 gene in 5 human glioma cell lines by Southern blot analysis. The p16 gene was located in the 9p21 chromosomal region and homozygous deletion was detected in 4 of 5 (80%) human glioma cell lines and 5 of 15 (33%) clinical samples. We transfected the full-length human p16 gene into p16-null human glioma cell line, U251MG cells, using the plasmid vector pRc/CMV-p16 and evaluated the effect of p16 gene transfer on the growth suppression of malignant glioma cells. The transfection of p16 cDNA caused growth suppression through G1 cell cycle arrest in U251MG cells. We also examined the effect of p16 gene transfer on the chemosensitivity to cis diamminedichloroplatinum II (CDDP), 1-(4-amino-2-methyl-5-pyrimidinyl)methyl-3-(2 chloroethyl) -3-nitrosourea hydrochloride (ACNU), and 5'-azacytidine (AZC). We did not detect any change in them after p16 gene transfer. These results might suggest that deletion of p16 genes promoted unrestrained growth in human glioma but has no relationship to the chemosensitivity to CDDP, ACNU and AZC. PMID- 9216648 TI - The inhibitory effect of selenium on induction of tetraploidy by dimethylarsinic acid in Chinese hamster cells. AB - Arsenic is a human carcinogen. On the other hand, selenium supplementation can inhibit induction of carcinogenesis by chemical carcinogens. The effect of selenium compounds on the cytotoxicity of dimethylarsinic acid (DMA) and on the induction of tetraploidy by DMA were studied using Chinese hamster V79 cells. Two selenium compounds were tested, sodium selenite and trimethylselenonium iodide (TMSeI). Trimethylselenonium is a major excretory product of selenite metabolism. The cytotoxicity of sodium selenite was 1000-fold greater than that of TMSeI. The cytotoxicity of DMA was about the same as that of TMSeI. The mitotic index for DMA administration was increased by these selenium compounds at low concentrations and decreased by them at high concentrations. The tetraploid index for DMA decreased with increasing concentrations of these selenium compounds. Tetraploidy is a form of aneuploidy, and aneuploidy is known to induce carcinogenesis. The finding that selenium inhibited induction of tetraploidy by DMA may yield clues to the role of selenium in the chemoprevention of carcinogenesis by chemical carcinogens. PMID- 9216649 TI - Evaluation of the IMMULITE BR-MA and CEA assays and comparison with immunoradiometric assays for CA15-3 and CEA in breast cancer. AB - This study was designed to evaluate the performance of a new automated assay system, the IMMULITE Automated Immunoassay Analyzer in comparison with more commonly used IRMA assays for measuring circulating tumour marker levels in breast cancer patients. The assays investigated measure the MUC1 mucin (CA15-3 antigen) or CEA. Serum samples from breast cancer patients with various stages of disease and from a group of normal individuals were analysed. Significant correlations were found between tumour marker levels measured using the IMMULITE BR-MA and the CA15-3 assays and between levels measured using the two CEA assay formats. Levels of IMMULITE BR-MA and CEA correlated with stage of disease suggesting that both are markers of tumour burden Levels in Stage III breast cancer patients were found to be significantly higher than those of normals using the IMMULITE system but not the IRMA assays. This would suggest a role for the automated system in the monitoring of breast cancer at an earlier stage than that at which tumour markers are routinely used. Elevated marker levels did not correspond to any particular site of metastasis however, a greater proportion of patients with multiple sites of metastasis had elevated levels compared with those with secondary disease at a single site. We conclude that the IMMULITE Automated Immunoassay Analyzer is of value in the routine surveillance of tumour marker levels. PMID- 9216650 TI - Retinoic acid combined with GM-CSF induces morphological changes with segmented nuclei in human myeloblastic leukemia ML-1 cells. AB - The effects of all-trans retinoic acid (ATRA), either alone or in combination with GM-CSF, on the induction of differentiation of a human myeloblastic leukemia cell line, ML-1, were investigated. ATRA alone caused only slight induction of NBT-reducing activity even at a high concentration (10(-7) M), but when combined with GM-CSF, it led to remarkable increase in the induction of NBT-reducing activity. Synergistic effect of both agents was also observed on morphological changes and the inhibition of cell proliferation. When ATRA or GM-CSF was used alone, neither parameter was changed substantially for long periods of up to 9 days. However, the combination of both agents induced remarkable morphological changes with segmented nuclei and also suppressed DNA-synthesizing activity. PMID- 9216651 TI - Controlled release of cisplatin incorporated into biodegradable poly-d, l-lactic acid. AB - Using a melt-pressing technique, we produced a small solid cylinder containing cisplatin (CDDP) embedded in poly-d, l-lactic acid (CDDP-PLA). The in vitro release of CDDP from the polymer was examined in an immersion system. CDDP was released continuously for more than four weeks with no initial burst. Drug distribution for CDDP-PLA was compared with CDDP solution (CDDP-SOL) by subcutaneous administration into the back of rats. In the CDDP-PLA group, a high concentration of CDDP was maintained in the subcutaneous tissues near the implants for 20 days. However, in the CDDP-SOL group, the concentration of CDDP was low by 10 days after drug administration. CDDP-PLA may become a useful tool in locoregional chemotherapy as a solid type of drug delivery system with longlasting release. PMID- 9216653 TI - 5-FU-induced apoptosis correlates with efficacy against human gastric and colon cancer xenografts in nude mice. AB - Apoptosis may be an important mechanism by which cancer cells are killed by certain agents. It is reported here that apoptosis is a key event in the killing of human tumor cells by 5-fluorouracil (5-FU) in vivo. Apoptosis induced by 5-FU was determined using two human gastrointestinal tumor xenografts serially transplanted into nude mice: a gastric carcinoma (SC-1-NU) highly sensitive to 5 FU and a colon carcinoma (Co-4) less sensitive to 5-FU. Apoptosis was assayed using the terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-biotin nick end labeling method in paraffin-embedded tissue sections, and by flow-cytometric analysis. Apoptosis-positive cells increased gradually during treatment. 24 hours after the initiation of 5-FU treatment a maximum, of 15.4% of the Co-4 cells were apoptotic. 48 hours after the initiation of 5-FU treatment, apoptosis was found in 34% of the tumor cells in the SC-1-NU strain. Flow-cytometry demonstrated the increase of S-phase fractions in both strains after the administration of 5-FU, and this coincided with the appearance of apoptotic-positive cells. Although the intrinsic. TS activities of two strains differed, TS activities were markedly suppressed in both strains immediately after the administration of 5-FU. Concentration of 5-FU in RNA (F-RNA) increased gradually in both strains, reaching a maximum 24 hours after the administration of 5-FU. These results suggest that apoptosis and inhibition of DNA synthesis induced by 5-FU are closely associated with its antitumor effect. PMID- 9216652 TI - Antitumor mechanism of intradermal administration of lipopolysaccharide. AB - We earlier demonstrated that 50% of the lethal dose of lipopolysaccharide (LPS) from Pantoea agglomerans given by the intradermal (i.d.) route is about 300 times greater than that given by the intravenous (i.v.) route, and that 400 micrograms/kg of LPS administered i.d. significantly suppressed metastasis whereas administered i.v., it did not. To learn the specific mechanism involved in this i.d. administration, the fate of LPS at the skin following administration and the concurrent production of endogenous tumor necrosis factor (TNF) in serum was examined. Histological observation following the i.d. administration of LPS (40 micrograms/kg) revealed neutrophiles in the skin 6 hours later. After 24 or 48 hours inflammatory cells were assembled at the site of injection. Endogenous TNF activity was found in the skin 24 hours after the injection and was significantly detectable even after 48 hours. Endogenous TNF was induced around tumor lesions of Meth A fibrosarcoma, MH134 hepatoma and Lewis lung carcinoma by treatment of LPS administered i.d. Taken together, these findings suggest that the antitumor activity of i.d. administered LPS results from the continuous supply of a small amount of this substance producing free TNF and activating inflammatory cells such as macrophages having membrane bound proTNF on their surface from the injected site to the tumor lesion for more than 48 hours. PMID- 9216654 TI - State of differentiation affects the response of endometrial adenocarcinoma cells to retinoic acid. AB - BACKGROUND: Patients with poorly differentiated endometrial cancers have a worse prognosis than patients with well-differentiated endometrial cancers. If poorly differentiated cells in endometrial cancers could be induced to differentiate, they would be more responsive to hormonal manipulation, and survival rates would be increased. We set up an in vitro model system to examine the effects of retinoic acid on human endometrial adenocarcinoma cells at three states of differentiation. METHODS: Cells were treated with pharmacological doses of 13-cis or all-trans retinoic acid (0.5 microM, 1 microM or 5 microM), and stained for mucins or actin filaments. RESULTS: Untreated undifferentiated (KLE) cells lack organized actin filaments and cytoplasmic mucins. Treatment with 5 microM retinoic acid caused some reorganization of actin filaments, but cytoplasmic mucins remained absent. Moderately differentiated (RL95-2) cells differentiated the most with retinoic acid treatment evidenced by a dramatic reorganization of actin filaments and an increase in cytoplasmic mucins. Untreated or treated well differentiated (Ishikawa) cells possessed well organized actin filaments and exhibit positive staining for cytoplasmic mucins. CONCLUSION: Retinoic acid causes cellular differentiation in less differentiated human endometrial adenocarcinoma cells. PMID- 9216655 TI - A newly synthesized bifunctional inhibitor, CKA1083, enhances adriamycin activity against human ovarian carcinoma cells. AB - BACKGROUND: A newly synthesized drug, CKA1083 ((S)-N-[2-(4-benzyloxy carbonylpiperazinyl)-1-(P-methoxybenzyl) ethyl]-N-methyl-N(5 isoquinolinesulfonamide)), has the same glutathione-S-transferase (GST)-binding site structure as W-77, a bifunctional inhibitor that enhances the cytotoxicity of Adriamycin for human ovarian carcinoma cells. We examined the effects of CKA1083 on the cytotoxicity of Adriamycin and the resistance of human ovarian carcinoma cells to this drug. MATERIALS AND METHODS: We used GST-pi transfected cells and Adriamycin-sensitive or -resistant cells of human ovarian carcinoma. GST-pi activity, the intracellular Adriamycin content, and the cytotoxicity of Adriamycin in these cell lines in the presence or absence of CKA1083 were measured and compared to the findings obtained with W-77 or verapamil. RESULTS: CKA1083 inhibited GST-pi activity in an uncompetitive manner and more strongly than W-77. It enhanced the cytotoxicity of Adriamycin for GST-pi transfected cells by about 3-times. Further, CKA1083 increased the intracellular Adriamycin content about 3-fold in two Adriamycin-resistant cell lines (NOS2AR and NOS3AR). CKA1083 (10 microM) reduced the IC50 of Adriamycin to 1/38 in NOS2AR cells and 1/21 in NOS3AR cells, and overcame Adriamycin resistance more effectively than both W-77 and verapamil. CONCLUSIONS: CKA1083 enhanced the antitumor effect of Adriamycin more than W-77 by inhibiting both GST activity and P-glycoprotein. PMID- 9216657 TI - Localization of mineralocorticoid receptor and 11 beta-hydroxysteroid dehydrogenase type II in human breast and its disorders. AB - Mineralocorticoid receptors have been detected in the normal human breast and breast cancers. The expression of mineralocorticoid receptor (MR) and 11 beta hydroxysteroid dehydrogenase type II (11sHSD2), which confers specificity on MR for aldosterone, was examined by immunohistochemistry in 114 samples from normal human breast and benign and malignant breast lesions in order to study its possible biological significance. MR and 11sHSD2 were immunolocalized in the ductal epithelium in 39/40 (98%) and 36/40 cases (90%) of normal breast, 21/22 (95%) and 15/22 cases (68%) of fibrocystic changes, and 11/11 (100%) and 8/11 (73%) cases of fibroadenoma, respectively. Cases positive for 11 sHSD2 also expressed MR but the patterns of expression varied greatly among examples of normal breast and benign breast diseases. There was a significant correlation between labeling indices of MR and 11sHSD2 in normal breast (p < 0.01) and in benign breast disease (fibrocystic change (p < 0.05) and fibroadenoma (p < 0.05)). In invasive carcinomas, immunoreactivity for MR and 11sHSD2 was detected in malignant cells in 32/41(78%) and 16/41(39%) cases. Both MR and 11sHSD2 labeling indices were significantly higher in invasive ductal carcinoma (22 cases) than invasive lobular carcinoma (19 cases) (p < 0.01). There was a significant correlation between labeling indices of MR and 11sHSD2 when analyzing all infiltrating carcinomas (p < 0.01), but not when assessing invasive lobular or invasive ductal carcinomas separately. These results indicate that the 11 sHSD2 enzyme generally colocalizes with the MR in the ductal epithelial cells of human breast, which may allow aldosterone to occupy its physiological receptor, and the expression of MR and 11sHSD2 appears to be related to ductal differentiation of breast carcinomas. PMID- 9216656 TI - Metallopanstimulin is overexpressed in a patient with colonic carcinoma. AB - BACKGROUND: Human metallopanstimulin (MPS-1) is a 9.4-kDa multifunctional ribosomal S27 nuclear "zinc finger" protein which is expressed in a wide variety of actively proliferating cells and tumor tissues. In this paper, we present a case of overexpression of MPS-1 in colon cancer tissues of a seventeen year old male. METHODS: Biopsies at the anastomosis and adjacent normal colonic mucosa were obtained by colonoscopy twelve months after right hemicolectomy for an ascending colon well differentiated adenocarcinoma. Immunohistochemical localization of MPS-1 protein was performed by using specific anti-MPS-1 antibodies directed against the N-terminal region of this protein. RESULTS: Immunohistochemistry demostrated an overexpression of MPS-1 in colonic mucosa crypts in the samples obtained at the anastomosis. In contrast, no expression of MPS-1 was observed in the adjacent normal mucosa. Histopathology performed with hematoxilin and eosin staining revealed focal crypt distortion and proliferation, but no carcinoma. CONCLUSIONS: In this case, the overexpression of MPS-1 was a more definitive evidence of malignant transformation than histology, as demonstrated by the clinical course of the disease. The results support the hypothesis that increased levels of tissue MPS-1 may correlate with a more aggressive behavior of colon malignancy. PMID- 9216658 TI - In vitro characterisation of soft tissue tumor chemosensitivity. AB - BACKGROUND: The benefit of performing chemotherapy on soft tissue sarcomas remains controversial. The present study deals with the in vitro characterisation of the influence of 3 antitumoral agents on the growth of 8 sarcoma cell lines. MATERIALS AND METHODS: Cell growth was monitored by means of the MTT colorimetric assay, which was further validated by a direct cell counting method. The three drugs tested included doxorubicin (ADR), cisplatin (DDP) and dacarbazine (DTIC). ADR was tested at 10(-5) M, 10(-6) M and 10(-7) M; DDP at 10(-5) M, 10(-6) M and 10(-7) M; and DTIC at 10(-3) M, 10(-4) M and 10(-5) M. A combination of the three drugs was also tested in order to ascertain whether a synergistic effect on cell growth inhibition could be obtained. A potential antineoplastic agent-induced influence on cell growth was determined 3 days after the addition of the diverse drug(s) to the culture media. The cell concentration was specifically adapted to each cell line. The 8 cell lines included 3 leiomyosarcomas, 1 malignant mixed Mullerian tumour, 3 rhabdomyosarcomas and 1 fibrosarcoma. RESULTS: The results show that of the three drugs tested, ADR was the most efficient in terms of the level of cell growth inhibition obtained and the number of cell lines whose growth was significantly inhibited. Of the three drugs, the least active was DDP. A significant synergistic effect was observed when the three drugs were added together to the culture medium. This synergistic effect was evident at the lowest doses tested for each drug. Whatever the histopathological type, the 8 cell lines exhibited a wide range of response to chemotherapy. CONCLUSIONS: The present study shows that the inhibition induced by 10(-7) M ADR, 10(-7) M DDP and 10(-5) M DTIC on sarcoma cell line growth is significantly more efficient than if each agent is tested individually. The in vitro methodology used here fits in with clinical reality because it enables sarcoma cell heterogeneity to be taken into account. PMID- 9216659 TI - Effect of G:C-->A:T transition, potentially arising from O6-guanine alkylation, in the transcription regulation of c-fos serum response element. AB - O6-meGs, if not repaired before cell undergo DNA synthesis, can cause erroneous pairing of thymine resulting in a G:C-->A:T transition, after the next DNA replication. It is known that the presence of O6-meG in promoter sequences inhibits the specific binding of transcription factors. Little is known on the effect of G:C-->A:T transitions on this binding. c-fos SRE was used as a model to study the effect of different G:C-->A:T transitions (at the positions -305, -306, -316, -319 and -320) in terms of SRE specific DNA-binding and functional ability to activate transcription of a reporter gene. The electromobility shift assay and a transient transfection assay were used. The G:C-->A:T transition at -320 caused 92% inhibition, while mutations at the positions -305, -306, -316 and -319 caused respectively 55, 43, 19 and 44% inhibition. The findings indicate that some G:C- >A:T transitions, potentially arising from O6-guanine methylation, can impair the regulation of c-fos transcription. PMID- 9216660 TI - Activation of resting T cells against the CA 72-4 tumor antigen with an anti CD3/CA 72-4 bispecific antibody in combination with a costimulatory anti-CD28 antibody. AB - The poor prognosis of advanced gastric carcinoma requires new therapeutic approaches. Among these, the specific activation of resting lymphocytes by bispecific antibodies may be promising. Here we describe the generation and function of a bispecific monoclonal antibody (bi-mab) with specificity for CD3 and for the tumor antigen CA72-4 (TAG72) on various gastrointestinal tumors. We established a hybrid/hybridoma by somatic fusion of two hybridoma lines secreting antibodies against CA72-4 and CD3 respectively and characterized its bimab OKT3/B72.3. In combination with costimulatory anti-CD28 antibodies resting peripheral lymphocytes could be activated specifically by bi-mab OKT3/B72.3 with T cell proliferation and IL-2 secretion. The bi-mab OKT3/B72.3 could also trigger the cytotoxicity of these T cells toward, CA72-4+ cells in vitro. Our results indicate, that bi-mab OKT3/B72.3 in combination with an anti-CD28 mab can redirect T cell cytotoxicity specifically against CA72-4+ tumor cells implicating a novel strategy for the specific immunotherapy of CA72-4+ tumors. PMID- 9216661 TI - Effect of intraperitoneally, intravenously and intralesionally administered monoclonal anti-beta-FGF antibodies on rat chondrosarcoma tumor vascularization and growth. AB - The growth and vascularization of many tumours has been reported to be associated with the overexpression of the potent mitogenic and angiogenic polypeptide basic fibroblast growth factor (beta-FGF). Consequently, it has been proposed that inhibition of beta-FGF action would prevent the growth of beta-FGF-dependent tumours. In this study, cell culture assays were established to assess the ability of mouse monoclonal DG-2 anti-beta-FGF antibodies to inhibit the mitogenic action of beta-FGF in vitro. Following in vitro characterisation, the monoclonal DG-2 antibodies were used to evaluate the role of beta-FGF in promoting the vascularization and growth of rat chondrosarcoma tumours. The effect the monoclonal anti-B-FGF antibodies had on tumour vascularization and growth in vivo were monitored using a 99m Technetium (99mTc)-labelled red blood cell procedure. The characterization studies confirmed that the DG-2 monoclonal antibody recognised beta-FGF and inhibited its mitogenic action on mouse Balb/c cells and bovine endothelial cells in vitro. When examined in vivo, intralesional administration of mouse monoclonal DG-2 antibody significantly inhibited rat chondrosarcoma growth and vascularization. However when the monoclonal DG-2 antibody was administered intraperitoneally or intravenously no attenuation of rat chondrosarcoma tumour vascularization or growth was observed. This report has confirmed the potential effectiveness of anti-beta-FGF antibodies in the regulation of tumour growth. It has also demonstrated that further studies on the pharmacokinetics of administered antibodies and their mode of delivery are required so that the effectiveness of such anti-growth factor immunotherapy can be assured. PMID- 9216662 TI - High sensitivity to hepato-tumorigenesis in hypocatalasemic C3H/C(s)b/Gen mice exposed to low doses of 252Cf fission neutrons and 60Co gamma-rays. AB - The sensitivity of hypocatalasemic C3H/C(s)b/Gen mouse (C(s)b) a mutant of the C3H/He mouse, for radiation tumorigenesis was studied following the irradiations of 252Cf fission neutrons (252Cf) and 60Co gamma-rays (60Co). Mice of both sexes at six weeks of age were irradiated once with 252Cf and 60Co at the doses of 0 200 cGy. As the control, C3H/HeN mice of both sexes (C3H) were also irradiated with 252Cf and 60Co. The mice were observed for 13 months after irradiation. The incidence of liver tumors in male C(s)b increased semilogarithmically from 1 to 50 cGy in the 252Cf-irradiated group. By 60Co irradiation, the tumor incidence of male C(s)b was highest at 6 cGy and then decreased with increasing radiation doses up to 200 cGy. In contrast, the incidence of liver tumors in male C3H steadily increased by 0-200 cGy of both 252Cf and 60Co irradiations. The glutathione peroxidase activity in the liver of the male was higher in C(s)b than that in C3H although catalase activity in C(s)b was lower than that in C3H. Significantly higher TBA value and lowered antioxidant activities were also observed in neoplastic liver foci in comparison with those of its adjacent liver in male C(s)b. Therefore, the implication of oxygen radicals in hepato tumorigenesis was strongly suggested and high sensitivity of C(s)b to low doses of radiation would be useful to investigate the mechanism of radiation tumorigenesis. PMID- 9216664 TI - Evidence of difference between cytotoxicity, cell cycle and morphonuclear effects of polyamines on sensitive and multidrug resistant P388 cell lines. AB - This paper reports studies on the influence of multidrug resistance (MDR) on the mechanism of polyamine toxicity. The effects of putrescine (PUT), spermidine (SPD) and spermine (SPM) on morphonuclear parameters and cell cycle were studied by means of digital cell image analysis. This reveals that only SPD and SPM condense chromatin inducing a strong decrease in the nuclear area and a cell cycle arrest in phase G2 in P388/s and the two MDR sublines. A significant difference was observed between the sensitivity of the two phenotypes, which confirms results obtained by means of a microculture tetrazolium test which showed that SPD and SPM were highly, but very differently, cytotoxic on sensitive and MDR sublines, unlike PUT, which was not toxic. This encourages us to study more thoroughly possible differences in polyamine metabolic enzymes and uptake in these cells, to enable us to acquire a better understanding of the impact of MDR phenotype on the polyamine pathway. PMID- 9216663 TI - Influence of benzo[a]phenothiazines on the element content of two tobacco tissue cultures differing in hormone requirement. AB - Cell proliferation and tumor formation are closely connected with hormone metabolism. We report the effect of four benzo[a]phenothiazines (12H benzo[a]phenothiazine (1), 5-oxo-5H-benzo[a]phenothiazine (2), 10-methyl-12H benzo[a]phenothiazine (3), and 6-hydroxy-5-oxo-5H-benzo[a]phenothiazine (4) on the growth and changes in the element compositions (Ca, Cl, Cu, I, K, Mg, Mn and Na) of auxin autotrophic and heterotrophic tobacco tissue cultures. The concentration levels of these ions were followed by means of reactor neutron activation analysis. PMID- 9216665 TI - Suramin is synergistic with vinblastine in human colonic tumor cell lines: effect of cell density and timing of drug delivery. AB - Suramin is a multi-targeted antiproliferative drug developed for the treatment of African trypanosomiasis but with potential efficacy for the treatment of human cancer. Cell growth inhibition was determined in vitro for three human colonic tumor cell lines using three different doses of suramin (50, 100 and 200 microM). At the lower suramin concentration cell growth was stimulated relative to control cultures in all three cell lines. At the higher dose which is at the upper end of the tolerable dose in humans suramin reduced cell numbers by greater than 50%. Inhibition of cellular proliferation was reduced relative to increases in cell plating density. Addition of vinblastine six and to a lesser extent 72 hours post suramin (200 microM) resulted in an inhibition of cell growth and/or toxicity that exceeded that which occurred as a result of exposure to either suramin or vinblastine alone. To investigate the possible mechanism by which suramin sensitizes cells to vinblastine we determined the effect of suramin on expression of the multidrug resistance (mdr1) gene. A decrease in mdr1 mRNA was evident in one colon tumor cell line and a slight decrease detected in a second line. The results establish that suramin is effective in controlling growth in colonic tumor cells and confirms that suramin activity is synergistic with other chemotherapeutics. The effect of suramin on MDR is of potential value that needs to be more thoroughly investigated particularly in cancers such as the those of the colon that are often drug refractory. PMID- 9216666 TI - C6 cells cross-resistant to cisplatin and radiation. AB - Malignant gliomas are relatively resistant to radiation and chemotherapy. To investigate whether cisplatin (cis-diamminedichloroplatinum(II), CDDP) causes resistance we pretreated C6 cells with 10(-6) M CDDP for 24 hours and then tested their sensitivity in a colorimetric assay. Pretreated cells developed resistance to CDDP (resistance factor 2.0) and radiation (survival after 9 Gy 60Co: 36.4% +/ 5.5 versus 28.6% +/- 5.2, p = 0.005). Glutathione levels of pretreated cells were higher (51.7 +/- 13.8 ng/mg protein) than in wt cells (40.4 +/- 13.2, p = 0.029). Addressing the mechanisms we established 4 wild type subclones with different CDDP sensitivities. However, cross-resistance to CDDP (survival: 60.7% +/- 3.5 versus 7.2% +/- 0.5, respectively p = < 0.001) and radiation (29.7% +/- 2.6 versus 12.9% +/- 0.8, p = < 0.001) could also be induced in a subclone showing involvement of mutation. These data suggest that CDDP can induce resistance mediated via induced mutation and increased GSH levels. PMID- 9216667 TI - Effect of alpha-tocopherol on cytotoxicity induced by UV irradiation and antioxidants. AB - The addition of DL-alpha-tocopherol (vitamin E) at the time of UV irradiation only marginally protects cells from UV-induced cytotoxicity. However, a protective effect of alpha-tocopherol emerged when it was added to the cells before UV irradiation, alpha-Tocopherol was progressively and dose-dependently incorporated into the cells. Washout experiments showed that the intracellular concentration of alpha-tocopherol decreased with an approximate half-life of 14 20 hours, due to the release from the cells and dilution by cell proliferation. Pretreatment of the cells with alpha-tocopherol significantly increased the resistancy against the cytotoxic action of UV irradiation and antioxidants such as sodium ascorbate, gallic acid, n-propyl gallate and caffeic acid. ESR spectroscopy showed that alpha-tocopherol enhanced the ascorbyl radical intensity, whereas it reduced caffeic acid radical intensity, without affecting the radical intensity of gallic acid and n-propyl gallate. Both control and treated cell lysates scavenged superoxide anion (generated by xanthine-xanthine oxidase reaction) and hydroxyl radical (generated by Fenton reaction) to a comparable extent. The present study suggests that the protective effect of alpha tocopherol might be derived from its incorporation into the cell membranes rather than its scavenging activity. PMID- 9216668 TI - Enhanced potency of daunorubicin against multidrug resistant subline KB-ChR-8-5 11 by a pulsed magnetic field. AB - Tumor cell resistance to many unrelated anticancer drugs is a major obstacle during cancer chemotherapy. One mechanism of drug resistance is thought to be due to the efflux of anticancer drugs caused by P-glycoprotein. In recent years, magnetic fields have been found to enhance the potency of anticancer drugs, with favorable modulation of cancer therapy. In this study, KB-ChR-8-5-11, a multidrug resistant (MDR) human carcinoma subline, was used as a model to evaluate the ability of pulsed magnetic fields (PMF) to modulate the potency of daunorubicin (DNR) in vivo and to determine the appropriate order of exposure to drugs and PMF using an in vitro cytotoxicity assay. Solenoid coils with a ramped pulse current source were used at 250 pulses per second for both in vivo and in vitro experiments. For the in vivo study, KB-ChR-8-5-11 cells were inoculated into thymic Balbc-nu/nu female mice. Treatment was begun when the average tumor volume reached 250-450 mm3. Treatment consisted of whole body exposure to PMF for one hour, followed immediately by intravenous (i.v.) injection of 8 mg/kg DNR designated as day 0, and repeated on days 7 and 14. Among the various groups, significant differences in the tumor volume were found between PMF + saline and PMF + DNR groups (p = 0.0107) at 39 days and 42 days (p = 0.0101). No mice died in the PMF alone group, and no toxicity attributable to PMF was found during the experimental period. For the in vitro studies, the sulforhodamine blue (SRB) cytotoxicity assay was used to determine the effect of the sequence which cells are exposed to PMF and/or DNR. Cells were exposed to PMF either before (pre-PMF) or after (post-PMF) drug was added. Results showed that the IC50 was significantly different between controls and pre-PMF + DNR groups (P = 0.0096, P = 0.0088). The IC50 of the post-PMF + DNR group was not found to be significantly different from control groups. Thus, the data in this report demonstrates that PMF enhanced the potency of DNR against KB-ChR-8-5-11 xenograft in vivo, while the efficacy of DNR was potentiated in vitro by PMF exposure only when PMF exposure occurred in the presence of drug. The data in vitro suggest that the mechanism by which PMFs modulate DNR's potency may be by inhibition of the efflux pump, P-glycoprotein. Further work to determine conditions for maximum modulation of drug potency by PMFs is warranted. PMID- 9216670 TI - The effects of cantharidin analogues on xanthine oxidase. AB - Norcantharidin[3], the demethylated product of cantharidin[1] has been used for the treatment of hepatoma, carcinomas of esophagus and gastric cardia, leukopenia and hepatitis. Since the enzyme xanthine oxidase is involved in the diseases mentioned above, and the reactive oxygen species produced by the enzyme induces DNA damage and oxidative damage of tissues, fourteen cantharidin analogues and cantharidimide derivatives were tested for their effects on xanthine oxidase. The results showed that these compounds, listed in Figure 1, displayed very weak inhibitory effects on xanthine oxidase. Contrary to expectation, disodium cantharidate [2], Norcantharidin [3], dehydronorcantharidin [4], disodium dehydronorcantharidate [5], N-(2-pyridyl) cantharidimide [12], N-(3pyridyl) cantharidimide [13] and N-(4-pyridyl) cantharidimide [14] showed a slight stimulating effect on xanthine oxidase at several concentrations. PMID- 9216669 TI - Apoptosis of androgen-independent mammary and prostate cell lines induced by topoisomerase inhibitors: common pathway of gene regulation. AB - New treatments for hormone-independent tumor are urgently needed since androgen dependent cancer cells eventually progress to -independent cells after hormonal manipulation. In the present study, etoposide and camptothecin were used to induce proliferation-dependent death of these cells. Each of the agents, at the doses used, induces the apoptosis of AT-3, CS 2, and TSU-pr1 cells based upon the temporal sequence of DNA fragmentation, morphologic changes and loss of cell viability. Northern blot analysis was used to identify a series of genes whose expression is enhanced during the apoptotic pathway. During the apoptotic process induced by the agents, expression of cyclin-dependent kinase inhibitor p27 increased. Flow cytometric analysis showed that the treatment resulted in a block in G2/M of the cell cycle. These results demonstrate that these cells retain the ability to undergo apoptosis by etoposide and camptothecin, and cyclin-dependent kinase inhibitor plays some role during apoptotic pathway. PMID- 9216671 TI - In vitro bioeffects of the antiestrogen LY117018 on desmoid tumor and colon cancer cells. AB - BACKGROUND: Clinical and experimental evidence suggest that estrogen has a role in the natural history of desmoid tumor (DT) and colorectal carcinoma. METHODS: The biological effects of LY117018, a nonsteroidal antiestrogen benzothiophene derivative, were assessed on a human adenocarcinoma cell line (HCT8 cells), and on DT cells and colorectal cancer derived fibroblasts in primary culture. RESULTS: LY117018 inhibited cell proliferation and collagen type I synthesis in DT cells. The compound also reduced cell growth in HCT8 cells and colorectal cancer fibroblasts. Binding experiments revealed the presence of estrogen binding sites in DT cells and frozen tissues but LY117018 did not displace [3H]17 beta E2 binding to DT cells. CONCLUSIONS: Present results demonstrate that LY117018 inhibits epithelial and fibroblastic colon cancer cells proliferation and proliferation and differentiation of desmoid cells in vitro. The lack of displacement of [3H]17 beta E2 binding to desmoid cells by LY117018 suggests the existence of distinct LY117018 binding sites. PMID- 9216672 TI - Role of apoptosis, proliferating cell nuclear antigen and p53 protein in chemically induced colon cancer in rats fed corncob fiber treated with the fungus Pleurotus ostreatus. AB - The role of apoptosis, proliferative cell nuclear antigen (PCNA) and p53 protein in the preventive effects of dietary fiber treated with the fungus Pleurotus ostreatus on rat-colon tumorigenesis was studied. Tumors were induced by five subcutaneous injections of 1,2-dimethylhydrazine (DMH), 20 mg/kg rat, once a week. Rats were fed a semi-synthetic fiberfree diet (control) or a high-fiber diet (15%) derived from corncob treated or non-treated with the fungus. The rats we sacrificed 24 weeks after the first carcinogenic injection. The fungus treated corn-cob significantly decreased tumor incidence (to 26%) as compared to 44% and 57% in the other dietary groups. The apoptotic index (AI) significantly decreased in malignant tissue as compared to non-tumorous tissue. PCNA and cytoplasmic content of p53 protein exhibited an increasing trend in malignant tissue as compared to benign tissue (at 15% and 18%, respectively). The fungus-treated corncob significantly increased the content of p53 in the cell cytoplasm (to 33%) and its serum levels in tumor-bearing rats (to 38%). The cellular concentration of PCNA decreased to 61% in tumors obtained from rats fed the fungus-treated corncob as compared to controls. A high positive correlation was found between tumor grade and p53 protein in the serum (r = 0.97) or in the cell cytoplasm (r = 0.77) and between tumor grade and PCNA (r = 0.81). An inverse relationship was found between tumor grade and AI (r = -0.63). We found that 15% of corncob fiber alone seems not to be enough to prevent chemically induced tumorigenesis. The corncob fiber (15%) treated with the fungus had a significant protective effect against DMH-induced rat colon cancer, even at 15% and this effect was accompanied by the activation of some cellular mechanisms such as apoptosis, PCNA and p53 protein activation. Incubation of corncob with the fungus Pleurotus os, increased the dietary fiber content up to 78%. Thus corncob inhibits colon cancer development, and, therefore, may considered of potential use to the public. PMID- 9216673 TI - Antiproliferative effects of an organic extract from the marine diatom Skeletonema costatum (Grev.) Cleve. Against a non-small-cell bronchopulmonary carcinoma line (NSCLC-N6). AB - An organic extract of the marine diatom Skeletonema costatum was studied in vitro for its effect on asynchronous cells of a human non-small-cell bronchoplumanory carcinoma line (NSCLC-N6). Cell growth appeared to be inhibited in the G1 phase of the cell cycle, and kinetic studies in pretreated cells showed that this growth arrest was irreversible. These events are related to a terminal maturation induction. PMID- 9216674 TI - Cytogenetics of parametrial leiomyoma. AB - The cytogenetical findings from a parametrial leiomyoma are presented. The results, together with those of five previously presented cases, show obvious differences when compared to the chromosomal findings in uterine myomas. Ontogenic factors are proposed to be causative for the cytogenetical differences. PMID- 9216675 TI - p53 protein expression in breast carcinomas. Comparative study with the wild type p53 induced proteins mdm2 and p21/waf1. AB - The aim was to investigate the pattern of expression of p53 protein and two wild type (wt) p53-induced proteins (mdm2 and p21/waf1), as an indirect way of assessing p53 gene status in breast carcinomas. Formalin-fixed paraffin embedded tissue from 102 cases of breast carcinomas comprising mostly ductal carcinomas (88 cases) was stained by immunohistochemistry for p53, mdm2 and p21/waf1 proteins. We found p53, mdm2 and waf1/p21 protein expression in 33/102, 20/102 and 38/102 breast carcinomas, respectively. Parallel p53/mdm2 protein expression was found in 9 cases. Five were also p21/waf1 positive. Discordant p53+/ mdm2 protein expression was found in 24 cases. Nine were p21/waf1 positive and the remaining fifteen were p21/waf1 negative. The patterns mdm2+/p53-/p21- and p21+/p53-(+)/mdm2- were found in 6 and 20 cases, respectively. Parallel p53/mdm2/p21 protein expression may represent breast carcinomas with wt p53 gene since mdm2 and p21 proteins are inducible by wt p53 gene. In these cases p53 protein expression may be due to stabilisation to mdm2 protein. This could be important in the pathogenesis of these cases since mdm2 may deregulate the p53 dependent growth suppressive pathway. Discordant p53+/mdm2-/p21- protein expression may represent breast carcinomas with p53 gene mutations unable to activate expression of mdm2 and p21 proteins. Breast carcinomas with p53+/mdm2/p21+ protein expression may have either wt p53 with deregulated mdm2 gene expression or mutated p53 gene with p53-independent p21 expression. Cases with only mdm2 expression may represent tumours with mdm2 gene amplification or overexpression and cases with only p21 expression may reflect p53-independent regulation of p21 protein. PMID- 9216676 TI - Glutathione derivatives enhance adriamycin cytotoxicity in a human lung adenocarcinoma cell line. AB - We evaluated the effects of a panel of glutathione derivative (S-butyl, S-decyl, S-ethyl, S-heptyl, S-hexyl; S-methyl, S-nonyl, S-octyl, S-propyl and S-pentyl glutathiones) on glutathione-S-transferase activity in the cell lysates of a human lung cancer, PC-9. Glutathione derivatives inhibited glutathione-S transferase activity in PC-9 cell lysates by up to 67%. When PC-9 cells were incubated with the IC50 concentration of adriamycin (200 nM) and with nontoxic concentrations (1 microM) of the glutathione derivatives, cytotoxicity ranged from -20% to +55% of the control levels. Enhancement of adriamycin toxicity by glutathione derivatives was significantly correlated with the inhibition of glutathione-S-transferase activity. S-decyl-glutathione, which was one of the most potent inhibitors of glutathione-S-transferase activity, significantly enhanced the adriamycin-induced antitumor effect in vivo. Findings suggest that some glutathione derivatives, including the S-decyl, S-octyl, and S-hexyl glutathiones, enhance adriamycin-induced cytotoxicity in part by inhibiting glutathione-S-transferase and that these agents may be useful as chemosensitizers for adriamycin therapy. In conclusion, the present results showed that some glutathione derivatives enhanced sensitivity of tumor cells to ADR by inhibiting GST activity. The use of BSO and EA as sensitizers to chemotherapy is currently being evaluated in clinical trials. The present data suggest that the use of GSH derivatives to modulate GST activity may improve the response to ADR. PMID- 9216677 TI - Enhancement of tumor associated antigen expression during the regression phase of xenogenized tumor cell growth in vivo. AB - Rat fibrosarcoma cells infected with Friend leukemia virus (FV-KMT-17) grow for a short time and then regress spontaneously in syngeneic hosts. This regression was caused by immunological mechanisms, because the tumor cells were renogenized. In this study, we have tried to find out whether tumor-associated antigen (TAA) expression in these xenogenized tumor cells can be modulated by xenogenization. FV-KMT-17 cells (1 x 10(7)), which were subcutaneously transplanted into ten rats, spontaneously regressed after temporary growth. All rats which rejected FV KMT-17 cells showed strong resistance to rechallenge with KMT-17 (1 x 10(6)) cells. To reveal the chronological modulation of TAA and virus-associated antigen (VAA), a single-cell suspension was obtained from the subcutaneous tumors and expression of these antigens was chronologically measured. TAA, termed CE7 antigen, was examined by anti-CE7 monoclonal antibody (MoAb) and VAA was examined by anti-FK1 MoAb which recognizes the FV env gene product (gp 70). Expression of VAA was not modulated through either the progression or the regression phase, but expression of TAA was strongly enhanced in the regression phase. These results show that enhancement of TAA expression occurs during the regression phase of FV KMT-17 growth in vivo and that TAA-expressing cells may stimulate anti-tumor immunity, resulting in acquisition of resistance against parental KMT-17 cells. PMID- 9216678 TI - The LEC (Long-Evans Cinnamon) rat as an animal model for bilirubin-induced tooth pigmentation. AB - The LEC (Long-Evans Cinnamon) rat is well known as a useful animal model for hepatic disease. We noticed the green pigmentation in incisors 2-3 weeks after acute hepatitis accompanied by severe jaundice. This study was undertaken to elucidate the cause of this phenomenon. Half of the pigmented teeth were examined by histopathological analysis and microradiographic analysis. Pigmentation was observed as a green stripe that ran parallel to the incremental line in the dentine. The microradiographic analysis disclosed enhanced permeability of the pigmented area as compared with other areas. The rest of pigmented teeth were dried, powdered and bilirubin was extracted with chloroform /methanol/acetic acid, 30:10:0.5; v/v under sonication. After centrifugation, the supernatant was collected and evaporated. The residue was dissolved in chloroform and its absorption spectrum measured after diazo reaction to reveal the presence of bilirubin. The spectral characteristics indicated the presence of bilirubin in the pigmented teeth. Thus, the LEC rat may be useful animal model for bilirubin induced tooth pigmentation. PMID- 9216679 TI - Correlation of MDR1 expression and oncogenic activation in human epithelial ovarian carcinoma. AB - The expression of the MDR1 gene has been shown to correlate with tumor aggressiveness and oncogenic activation both in experimental tumor models and in human clinical specimens In order to verify whether this association also takes place in ovarian carcinoma, we studied tumor samples from 39 patients by means of immunohistochemistry for the overexpression of P-glycoprotein (MDR1), nm23, c-erb B2 and p53. MDR1 (p = 0.023), nm 23 (p = 0.037) and c-erb-B2 (p = 0.015) were expressed significantly more in specimens from patients with advanced stage of disease. There were no differences in p53 expression between both groups of patients. Furthermore, we found a significant coexpression of MDR1 and nm23 (p = 0.028), and of MDR1 and c-erb-B2 (p = 0.0077). There was no association between the expression of the MDR1 gene and p53. These results parallel those previously reported by us for mammary carcinoma, and seem to indicate that expression of the multidrug resistance gene (MDR1) is inherent to the development of the malignant phenotype in several human tumors. PMID- 9216680 TI - Anti-tumor effect of lipopolysaccharide by intradermal administration as a novel drug delivery system. AB - We examined the antitumor effect of lipopolysaccharide extracted from Pantoea agglomerans, a Gram-negative bacterium, using intradermal administration on murine syngeneic tumors, Meth A fibrosarcoma, MH134 hepatoma and Lewis lung (LL) carcinoma. The latter two tumors are known to be relatively low in immunogenicity, highly metastatic and to have low sensitivity to biological response modifiers. Although the intradermal administration of LPSp had a significantly suppressive effect on the growth of all tumors, including seventy five percent of complete regression of mice bearing Meth A tumor, no complete regression was observed in MH134 or LL tumors. In combination with cyclophosphamide given once prior to the administration of LPS, however, the antitumor effects by intradermal administration of LPS were significantly augmented and there was complete regression in all types of tumors. Pretreatment by anti-tumor necrosis factor antibody reduced the effect exerted by LPS, suggesting that induced tumor necrosis factor might have a crucial role. Toxicity of intradermal administration of LPS was 230-380 times less than that by the intravenous route. Thus clinical application of LPS administered intradermally in combination with chemotherapeutics such as cyclophosphamide appears promising in terms of its antitumor effect as well as toxicity. PMID- 9216681 TI - Cytokinetic and morphologic differences in ovarian cancer cells treated with ET 18-OCH3 and the DNA-interacting agent, etoposide. AB - New antineoplastic agents with different cytotoxic mechanisms are of interest for their ability to overcome resistance to conventional DNA-interacting agents. Ether lipids are known to be active against ovarian carcinoma both in vitro and in vivo, and the cell membrane is believed to be the target of their antitumor activity. In this study we have investigated the different cytokinetic and morphologic responses of human ovarian carcinoma cells (BG-1) to one of the ether lipids (ET-18-OCH3) and to etoposide. Etoposide induced a significantly greater G2/M block. However, the proportion of the cycling cell fraction decreased significantly in cells treated by ET-18-OCH3 and induction of the hypodiploid traction was strongly correlated with reduction of the cycling cell fraction. On the other hand, the hyperdiploid fraction was found to correlate with reduction of the cycling cell fraction in etoposide treated cells. Despite the significant appearance of the hypodiploid fraction, apoptosis was not observed by DNA-gel assay. Microscopic study showed that the hyperdiploid fraction represented cells with multiple nuclei. These observations support the unique lethal effect of ET 18-OCH3 on ovarian carcinoma cells, distinguishing it from the action of a typical DNA-interacting agent. The membrane-targeted ether lipids deserve consideration for the future chemotherapy of ovarian carcinoma, perhaps in combination with the appropriate DNA-interacting agent. New antineoplastic agents with different cytotoxic mechanisms are of interest not only for their unique inhibitory properties but also for their potential of overcoming resistance to conventional DNA-interacting agents. Ether lipids are known to be active against ovarian carcinoma both in vitro (1, 2, 3) and in vivo (4, 5), and the cell membrane is believed to be the target of their antitumor activity. Etoposide, a DNA-interacting agent, is also active against human ovarian cancer cells in vitro (6) or in clinical trials either as a single agent (7) or in combination with cisplatin (8). We have reported that a cytotoxic dose of one of the ether lipids, ET-18-OCH3, induces a G2/M block in BG-1 human ovarian cancer cells, and also a hypodiploid fraction as shown on DNA analysis by flow cytometry (FCM) (9). The G2/M block was also observed in BG-1 cells following etoposide treatment (6). In the present study, we have investigated the differences in the cytokinetic and morphologic responses of BG-1 cells to ET-18-OCH3 and to etoposide. PMID- 9216682 TI - Antioxidant enzyme activity in murine hematopoietic bone marrow following treatment with interleukin 1 alpha: influence of tumor. AB - To determine whether non-hematologic tumors influence the bone marrow's antioxidant enzyme response to the radioprotective cytokine interleukin 1 alpha (IL-1), studies were undertaken using BDF1 and Balb/c mice bearing small, medium or large Lewis lung carcinoma (LLCa) or EMT6 mammary carcinoma tumors, respectively. Results demonstrated that, similar to nontumor-bearing mice, treatment of tumor-bearing animals with IL-1 was associated with a significant increase in marrow MnSOD activity. However, the duration of this elevated activity was reduced as tumor burden increased, and this reduction may have an impact on IL-1's ability to radioprotect tumor bearing animals, especially when tumor burden is large. In addition to cytokine-mediated responses, significant tumor-related influences on the marrow's antioxidant enzyme status were seen. Notably, it was observed that the presence of tumor was correlated with a marked suppression of antioxidant enzyme activity. Surprisingly, however, the pattern of enzyme suppression was found to differ between the two tumor models studied both in temporal onset and in the number of enzymes involved. In conclusion, the data obtained from these studies on tumor-bearing animals demonstrate that there are both cytokine-related and tumor-related influences which can effect the antioxidant enzyme status of the hematopoietic marrow-influences which may have the potential to alter the marrow's ability to tolerate free radical-generating events, both endogenous (i.e inflammation, infection) and exogenous (i.e. radiation, certain chemotherapeutic drugs) in origin. PMID- 9216683 TI - Effect of cysteine, N-acetyl-L-cysteine and glutathione on cytotoxic activity of antioxidants. AB - The effect of twenty amino acids on the radical intensity of four antioxidants (sodium L-ascorbate, sodium 5,6-benzylidene-L-ascorbate, gallic acid, caffeic acid) was investigated, using ESR spectroscopy. Methionine and methional did not significantly affect the radical intensity of these antioxidants. Methionine sulfoxide slightly enhanced the radical intensity of sodium ascorbate and sodium 5,6-benzylidene-L-ascorbate, but did not that of gallic acid and caffeic acid. Cysteine, N-acetyl cysteine and glutathione significantly reduced the radical intensity and cytotoxic activity of these antioxidants except for sodium 5,6 benzylidene-L-ascorbate. The other amino acids were inactive. The present study further supports that these antioxidants induce cytotoxicity via their pro oxidant action. PMID- 9216684 TI - Effect of copper and iron ions on cytotoxicity induced by ascorbate, gallate and caffeate. AB - Four antioxidants, sodium ascorbate, gallic acid, n-propyl gallate and caffeic acid, induced apoptotic cell death in human promyelocytic leukemic HL-60 cells. The effects of all these compounds were enhanced by CuCl2 or deferoxamine mesylate, an iron chelator, but were reduced by FeCl3. ESR spectroscopy showed that both CuCl2 and FeCl3 enhanced the ascorbyl radical intensity, but reduced the gallate and caffeate radical intensity. The present data demonstrate that copper and iron ions modify the cytotoxic activity of these antioxidants differently and their radical intensity is not the sole determinant of cytotoxic activity. PMID- 9216686 TI - Human papillomaviruses are not associated with renal carcinoma. AB - In an attempt to better understand the troubling issue of renal human papillomavirus (HPV) infection, fresh tumour specimens obtained by surgical extirpation from tumours were compared for the presence of HPV. All of the renal carcinoma samples were examined by PCR using two sets of consensus primers and the specific primer pairs for HPV 16, 18, and 33. None of the 28 carcinomas and 17 corresponding normal tissues were found positive for HPV DNA in any of the applied analyses. Our results suggest that HPV does not play any role in the development of renal carcinoma in the general population. PMID- 9216685 TI - Characterization of two newly established cell lines derived from squamous cell carcinomas of the oesophagus. AB - This study describes characteristics of two newly established cell lines (OSC-1 and OSC-2), derived from two oesophageal squamous cell carcinomas. Morphologically, OSC-1 cells and OSC-2 cells grew in epithelial cobblestone patterns with cells piling up to 4 cells. Ultrastructurally, both cell lines showed formation of desmosomes; however, tonofilaments were only formed by OSC-2 cells. Immunohistochemical investigations revealed coexpression of vimentin and cytokeratin in OSC-1 cells and OSC-2 cells. A cytokeratin subtype typical for mature squamous epithelia (cytokeratin 13) was expressed only in OSC-2 cells. OSC 1 cells showed tumour formation in nude mice, whereas OSC-2 cells did not. Cytogenetic analysis revealed that OSC-1 cells had a hyperdiploid karyotype and OSC-2 cells had a near-triploid karyotype. In both cell lines, the formation of multicellular spheroids could be induced. In conclusion, in comparison with OSC-2 cells the OSC-1 cells were characterized by a poorer degree of differentiation and by a more aggressive growth behaviour in vivo. PMID- 9216688 TI - p53 protein overexpression in laryngeal squamous cell papillomas. AB - We retrospectively investigated p53 protein immunoreactivity in 103 laryngeal squamous cell papillomas (LP) previously revealed to be human papillomavirus type 6 or 11 positive by in situ hybridization and/or the polymerase chain reaction. 21 LP failed to show any detectable level of p53 protein reactivity. In 45 cases only occasional strongly positive cells were observed in almost the whole thickness of the epithelium. In 26 LP, p53 protein immunoreactivity was found to be almost exclusively restricted to the basal epithelial cells. Finally, in 11 cases, basal cell layer immunoreactivity was accompanied by aggregates of p53 positive cells in the lower two thirds of the epithelium. This staining pattern was found predominantly in LP with atypical hyperplasia. We think that the observed patterns of p53 immunoreactivity in a majority of cases are a result of immunohistochemical detection of the stabilized wild-type p53 protein rather than the mutated p53 protein. PMID- 9216687 TI - Phosphatidylinositol 3-kinase mediates heregulin-induced growth inhibition in human epithelial cells. AB - Activation of phosphatidylinositol 3-kinase (PI-3K) has been shown to be critical for heregulin induced mitogenesis of human breast epithelial cells. However, the inhibitory effects of HRG have not been linked to PI3K activation. The purpose of these experiments was to determine the effects of heregulin beta 1 (HRG) on growth and PI3K activation of a nonneoplastic human mammary epithelial cell line, 184B5. The results of these experiments indicated that low concentrations of HRG increased the growth of 184B5 cells three fold, while high concentrations inhibited growth by 50%. HRG at all concentrations tested increased tyrosine phosphorylation of erbB3. Similarly, HRG at all concentrations stimulated the association of PI3K with erbB3. Wortmannin, an inhibitor of PI3K enzymatic activity, reversed both the inhibitory and stimulatory effects of HRG on incorporation of [3H] thymidine into DNA. We conclude that both the growth inhibitory and stimulatory effects of HRG in nonneoplastic human mammary epithelial cells are mediated by activation of PI3K. PMID- 9216689 TI - Effect of blocking TNF on IL-6 levels and metastasis in a B16-BL6 melanoma/mouse model. AB - We studied the relationship between tumour necrosis factor (TNF) and interleukin 6 (IL-6) levels, and the metastatic process in C57BL/6 mice after intravenous inoculation of B16-BL6 melanoma cells. Bioactive TNF was not detectable in the sera of inoculated mice, but these animals did show higher TNF levels following intraperitoneal challenge with lipopolysaccharide (LPS) compared to control animals. Serum IL-6 levels were increased in inoculated animals. Injection of a hybrid molecule (p55-sf2) composed of the human p55 TNF receptor extracellular domain coupled to a human constant region backbone, decreased serum TNF (after LPS challenge) and IL-6 levels in inoculated animals. Lung metastases at 7-14 days were reduced, compared to human IgG-injected control animals, but this effect was lost at day 21 postinoculation. The results suggest that the reduction in the number of metastases may be related to the effect of blocking TNF activity. PMID- 9216690 TI - Hyperthermia and verapamil inhibit the growth of human colon cancer xenografts in vivo through apoptosis. AB - BACKGROUND: The long-term goal of this work is to develop a new therapeutic regimen for the treatment of colon cancer in humans which will include hyperthermic intraperitoneal perfusion of verapamil as an alternative to administration of chemotherapy. METHODS AND RESULTS: Hyperthermia and verapamil caused a significant inhibition of the growth of human colon cancer (HT-29) xenografts. Both apoptosis detection assays, TUNEL and H and E staining, have shown that approximately 50% of human colon cancer cells underwent apoptosis after hyperthermia and verapamil administration. The TUNEL assay has demonstrated that DNA strand breaks appeared fairly rapidly and maximum breakage occurred at 2 hours after the treatment. Histopathological assay has showed maximum apoptotic morphological changes at 12 hours after treatment. CONCLUSION: Thus, the results of our in vivo experiments confirmed our previously obtained in vitro data concerning the significant ability of the combination of hyperthermia and verapamil to inhibit human colon cancer cell growth through programmed cell death. PMID- 9216691 TI - Effect of combination therapy with UFT plus cisplatin (UFTP) on the survival of mice in the experimental model for wide-spread metastasis in the peritoneal cavity of gastrointestinal cancer using colon 26 PMF-15 cells. AB - The prognosis of unresectable advanced gastric cancer patients, especially with wide-spread metastasis in the peritoneal cavity, is very poor. In such cases, only a few treatment methods are available, and chemotherapy as a systemic therapy is often selected. We recently devised an experimental model for wide spread metastasis in the peritoneal cavity of gastrointestinal cancer, by intraperitoneally implanting colon 26 murine carcinoma PMF-15 cells. When the control mice were autopsied, a number of tumor nodules were formed in their mesenterium, pancreas, liver, etc. and pools of ascites were also occasionally seen. Using this model, oral UFT (combining tegafur and uracil at a molar ratio of 1:4) plus cisplatin (UFTP regimen) prolonged the survival of mice and maintained these mice in relatively better condition than with continuous infusion of 5-fluorouracil plus cisplatin (FP regimen). Since UFT can be used orally, patients receiving UFTP therapy are not required to stay in a hospital for long periods, but rather, are intermittently hospitalized for short periods of time. Better QOL and prolonged survival highlight the potential clinical usefulness of the UFTP therapy. PMID- 9216692 TI - Maintenance chemotherapy with oral treosulfan following first-line treatment in patients with advanced ovarian cancer: feasibility and toxicity. AB - PURPOSE: To evaluate the feasibility and toxicity of maintenance oral treosulfan chemotherapy for ovarian cancer patients after surgical treatment and response to first-line chemotherapy. PATIENTS AND METHODS: Thirty-nine patients were entered onto this trial. This was a pretreated patient population. The pretreatment consisted of radical surgery and chemotherapy. The treatment that immediately preceded oral treosulfan was standard-dose platin-based chemotherapy. Daily oral treosulfan was administered at a dose of 1250 mg for 5 consecutive days every five weeks for at least three cycles. All patients started daily oral treosulfan while in complete remission. RESULTS: A total of 322 cycles of oral treosulfan was administered, with a median of 6 cycles (range 3-24). Treosulfan in this schedule was generally well tolerated. The major toxic effects were leukopenia and thrombocytopenia, which, however, were manageable and rapidly reversible. There were no episodes of bleeding or leukopenic fever. No anti-emetic drugs were required. Alopecia was not observed, 20 patients had progressive disease (after 3 6 months: n = 8, after > 6 months: n = 12). The median survival for all patients was 24 (range 9.44+) months, and median time to progression 8 (range 3-24) months. CONCLUSIONS: Maintenance oral treosulfan was well tolerated in this pretreated patient population. In an attempt to further improve overall survival in ovarian cancer patients, prospective random assignment trials will be necessary to determine the benefit of this approach. PMID- 9216693 TI - Localization of aberrant messenger RNA of epidermal growth factor receptor (EGFR) in malignant glioma. AB - Human gliomas occasionally show rearrangements with deletions (exons II to VII) in the epidermal growth factor receptor (EGFR) gene, resulting in the expression of aberrant EGFR mRNA. This abnormality of EGFR gene expression is closely related to the malignancy of glioma. However, this EGFR gene abnormality has not been demonstrated directly in the glioma cells themselves, as gliomas consist of heterogeneous tissue components. In this study, we used in situ hybridization (ISH) to detect aberrant EGFR mRNA in the tumor cells in 26 human gliomas and 19 human glioma xenografts. We used digoxigenin (DIG)-labeled antisense oligonucleotide probes for ISH, which demonstrated aberrant EGFR mRNA in 2 of the 26 gliomas and in 3 of the 19 human glioma xenografts. ISH with an aberrant EGFR specific probe (oligo-PA) revealed that EGFR mRNA was absent from multinucleated giant cells and from proliferating endothelial cells, but this transcript was present in small glioma cells. Identical aberrant EGFR mRNA was confirmed in these glioma and human glioma xenografts by Southern blotting. Northern blotting, and by reverse transcription-polymerase chain reaction (RT-PCR). These findings suggest that small tumor cells specifically express the aberrant EGFR mRNA in certain high grade gliomas. PMID- 9216694 TI - Differential expression of apoptosis associated genes bax and bcl-2 in ovarian cancer. AB - The prognostic value of various molecular markers, which adequately account for the tumor biology and disease behaviour of ovarian cancer, is still unclear. Recent studies have focused on the role of genes regulating the balance between proliferation and cellular suicide, apoptosis. In the present study, tumor tissue from 215 patients with ovarian cancer was immunohistochemically analysed for Bax- and Bcl-2-expression. There was an association between Bcl-2-expression (30%) and factors of favourable prognosis. In contrast, Bax-expression (47%) was related to bad clinical outcome, especially in cases without concomitant Bcl-2-expression. In patients with Bcl-2-positive/Bax-negative tumors, overall survival was significantly longer (p = 0.0379) than in patients with Bcl-2- and Bax-negative tumors. Respectively, expression of Bax without Bcl-2-expression was correlated with bad clinical outcome (p = 0.033). The difference in overall survival was most striking (p = 0.0007) between patients with Bax-positive/Bcl-2-negative and Bcl-2-positive/Bax-negative tumors. This could also be demonstrated for the various subgroups of different tumor grade and stage. It may be speculated, that alteration of the Bax/Bcl-2-balance may influence the clinical course by deregulation of programmed cell death and altered sensitivity to chemotherapy. PMID- 9216695 TI - Reduced expression of molecules of the cadherin/catenin complex in the transition from colorectal adenoma to carcinoma. AB - E-cadherin is crucial to the intercellular adherens junctions which are involved in the organisation and maintenance of epithelial structure and suppression of tumour invasion. E-cadherin is associated with the actin cytoskeleton via cytoplasmic proteins, including alpha-, beta- and gamma-catenins, which together form the cadherin/catenin complex. To evaluate changes of the molecules of the cadherin/catenin complex in colorectal carcinogenesis, seventy-four sporadic adenomas, samples of histologically normal epithelium adjacent to 65 adenomas, and 52 carcinomas arising in adenomas were investigated by immunohistochemistry. All normal epithelial cells showed a uniform membranous staining pattern for E cadherin, alpha-, beta-, and gamma-catenin. Decreased expression of all 4 proteins occurred in parallel in adenomas and carcinomas (in all cases, p < 10( 5). Decreased expression of the cadherin/catenin complex in adenomas was associated with increasing severity of dysplasia (p < 0.001, for E-cadherin, alpha-, and gamma-catenin, p < 0.005 for beta-catenin). Carcinomas displayed significantly reduced expression of the cadherin/catenin complex compared with their associated adenomas (all p < 0.001). The results directly confirm that colorectal tumour progression and invasion is associated with disruption of the cadherin/catenin complex and suggest that the genetic changes and transcriptional modulation of catenins underlying this progression may affect all members of the complex. PMID- 9216696 TI - Coexpression of VEGF and bFGF in human epidermoid lung carcinoma is associated with increased vessel density. AB - Tumor specimens from 84 patients with untreated epidermoid lung carcinomas were analysed immunohistochemically for the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), tumor cell proliferation (PCNA index) and tumor vascularity (vessel density). The purpose of this study was to find out whether differences in tumor cell proliferation and tumor vascularity might be associated with differential angiogenic growth factor expression. The present results indicate that the proliferation of the tumors is closely related to their expression of VEGF, but not for bFGF. The PCNA labelling index in VEGF positive tumors (VEGF+/bFGF- or VEGF+/bFGF+) was significantly higher than that in VEGF negative tumors (VEGF-/bFGF- or VEGF-/bFGF+; Wilcaxon rank sum test, p < 0.0001). To investigate whether VEGF or bFGF is involved in lung tumor angiogenesis, the data of VEGF and bFGF expression were correlated with vessel density. It was found that the expression of VEGF and bFGF were associated with increment of vessel density, however, vessel density was significantly increased only when VEGF and bFGF were coexpressed (p < 0.02). It is suggested that VEGF and bFGF might act cooperatively in the neovascularization of human epidermoid lung carcinomas. PMID- 9216697 TI - Elevated levels of serum and plasma metalloproteinases in patients with gastric cancer. AB - BACKGROUND: Levels of serum and plasma metalloproteinases, especially type IV collagenases are important factors related to metastasis and invasion in various human cancer. The clinical significance of serum matrix metalloproteinase 2 (MMP 2, 72 kDa type IV collagenase) and plasma matrix metalloproteinase 9 (MMP-9, 92 kDa type IV collagenase) was evaluated as possible tumor markers in gastric cancer. MATERIALS AND METHODS: The precursor form of MMP-2 (proMMP-2) in serum and the precursor form of MMP-9 (proMMP-9) in plasma were examined prior to surgery on 70 patients with gastric cancer, one-step sandwich immunoassay (EIA) was used and serum carcinoembryonic antigen (CEA) levels were also examined in the same patients. Normal sera and plasma samples obtained from healthy individuals without cancer were used as controls. RESULTS: ProMMP-2 levels in patients (mean +/- standard deviation) with gastric cancer (602 +/- 200 micrograms/l) were significantly higher than levels (542 +/- 80 micrograms/l) in sera from 70 healthy individuals (P < 0.02). Plasma proMMP-9 levels (119 +/- 232 micrograms/l) in patients with cancer were also significantly higher than those (37 +/- 13 micrograms/l) in plasma (P < 0.004). On the other hand, there was no significant correlation between CEA and proMMP-2 levels, and between CEA and proMMP-9 levels. Neither proMMP-2 levels nor proMMP-9 levels significantly related to clinicopathologic features. CONCLUSIONS: Serum proMMP-2 levels and plasma proMMP-9 levels may serve as tumor markers, independent from known factors and CEA. PMID- 9216698 TI - DNA ploidy and HPV subtypes in cervical smears of HIV-sero-positive and negative patients. AB - Epidemiological studies have demonstrated that cervical HPV infection and precancerous lesions of the cervix are more common in HIV-seropositive patients. However little is known about the natural history of these lesions in this population. In the present study cervical smears from 36 patients, 18 HIV seropositive women and 18 matched controls were evaluated with the aim of quantifying morphological alterations and to evaluate DNA ploidy and HPV subtypes. Cervical lesions in HIV-seropositive patients were diploid in 50% of the cases compared to 25% in controls. The only HPVs identified by ISH were types 16/18 and no significant differences were observed in the control population. In contrast, cytological evidence of HPV infection and dysplastic changes was greatly increased in smears from HIV patients compared to HIV-seronegative women. Less than 5% of the cells showed HPV associated changes in controls while 10% to 30% of the cells were affected in HIV-patients. We suggest that the Papanicolaou test should be effective for detecting cervical disease and for a close follow-up of this population. Moreover, while additional studies with larger population groups and different population bases are needed, these findings are suggestive of the possible use of morphological criteria for the identification of HIV seropositive subjects. PMID- 9216699 TI - Para-aortic lymph node recurrence in patients with cervical cancer treated with postoperative irradiation to the pelvis. AB - A retrospective analysis of para-aortic lymph node (PALN) metastasis in patients with cervical squamous cell cancer who received prophylactic postoperative irradiation to the pelvis, was carried out. PATIENTS AND METHODS: Sixty-seven patients had no intra-pelvic lymph node (IPLN) metastasis (n 0), whereas 33 had IPLN (n 1). All patients received prophylactic external beam irradiation with a total dose of 50 Gy to the entire pelvis followed by intravaginal boost. Only 5 patients received a prophylactic PALN irradiation with a total dose of 30-50 Gy. RESULTS: No patients with n 0 developed para-aortic recurrence. However, 6 of 33 patients with n 1 developed PALNs recurrence but only 3 patients developed isolated PALN recurrence. There was no difference in survival between patients with PALNs irradiation and patients without PALNs irradiation. CONCLUSIONS: Routine prophylactic irradiation to PALNs is not necessary even for patients with IPLN metastasis except for the selected patients in the management of cervical cancer. PMID- 9216700 TI - Prognostic significance of immunophenotypes in adult lymphoblastic lymphomas. AB - Adult lymphoblastic lymphoma (LBL) can be of T-cell or B-cell lineage. However, the clinical significance of immunophenotypes is largely unknown. We conducted a retrospective study to compare T-cell LBL with its B-cell counterpart. Between 1983 and 1995, 50 adult patients were diagnosed as LBL at National Taiwan University Hospital. Twenty-seven patients (T-LBL:20 and B-LBL:7) had adequate clinical information and formed the basis of final analysis. Pertinent characteristics, including sex, age, and lymphoma stages of these two groups of patients were identical. Detailed clinical features were compared. Systemic involvements of lymphoma were similar except that T-cell LBL had significantly more mediastinal involvement (T:B = 70%:14.3%, p = 0.011). CNS involvement was high in both groups (T:B = 50%:28.6%, p = NS). B-cell LBL had a better overall survival than T-cell LBL, although the survival benefit became less significant after 30 months. The median survival of T- and B-cell LBL was 8 and 31 months, respectively. Both groups taken together, patients who had received prophylactic cranial irradiation had a better overall survival (p < 0.01). We suggest that: a) B-cell LBL has a relatively favorable prognosis than T-cell LBL, at least in the initial 2 to 3 years; b) except for mediastinal involvement, the clinical presentation of T- and B-cell LBL appears to be similar; c) treatment policy, such as the need of prophylactic cranial irradiation, of these two groups may also be similar. PMID- 9216702 TI - Gastric cancer and flow cytometry: the beginning of a promising match. AB - Gastric cancer (GC) is still a diffuse and aggressive disease in many countries where its frequency has not drastically decreased as in the USA. Furthermore, except in Japan and few other countries where the health care system has implemented effective strategies to diagnose the disease very early, GC therapy is unsatisfactory because of the advanced stage at which it is disclosed. Recent advances in the understanding of GC, resulting from identification of Helicobacter pylori as a critical factor in its development, seem not to have changed the situation. Studies on GC have often adopted flow-cytometry to investigate many parameters mostly related to prognosis. Unfortunately, the results obtained with this important technique have been disappointing and diverse, which has caused a fair amount of skepticism as to its role. Starting from a critical review of the vast Literature on the topic, this paper discusses a re-evaluation of the role of flow-cytometry in the fight against GC. PMID- 9216701 TI - A case of malignant schwannoma with overexpression of multidrug resistance gene (MDR1) after chemotherapy. AB - A 19-year-old:female patient with malignant schwannoma in the right knee was treated by combination chemotherapy including lipophilic anticancer compounds (cyclophosphamide, doxorubicin, vincristine and dacarbazine). The tumor was radically removed after chemotherapy, but metastatic lesions were noted in the right inguinal nodes. The patient died due to the cachexic state six months after surgery. In human neoplasm, P-glycoprotein (P-Gp) encoded by human multidrug resistance gene MDR1 is known to be related with multidrug resistant phenotype. Northern blot analysis revealed apparent MDR1 expression in the metastatic lesion, while the primary lesion showed faint MDR1 expression detected by only reverse transcriptase-polymerase chain reaction. P-Gp positive tumor cells were immunohistochemically detected both in the metastatic lesion and the primary lesion. The P-Gp-positive tumor cells in the metastatic lesion showed anaplastic features with highly atypical nuclei. These results suggest that MDR1 overexpression is related to the multidrug resistance phenomenon in the malignant schwannoma with morphological differences. PMID- 9216703 TI - Differential implications of the oncogene-tumor suppressor gene complex in the geneses of 19 human neoplasias. Evidence in support of the steroid carcinogenesis hypothesis. AB - The cancer risk changes of 19 human neoplasias over time and space, as expressed in terms of the logarithm of age-adjusted incidence rate (log AAIR), were found to hold a linear correlation with each other--a finding suggesting that the distribution pattern of log AAIR data sets of 2 cancers, when plotted on a two dimension diagram, may show a good fitness to the chemical equilibrium model a product of the law of mass action. On the basis of the statistical analysis of the data, we reached the conclusion that the risk changes of a given neoplasia in space represents the function of the centripetal force of an activated oncogene and the centrifugal force of an inactivated tumor suppressor gene, both of which should cooperate with each other to create a thermodynamic equilibrium under the law of mass action. The purpose of this study was to test the contribution of the oncogene-tumor suppressor gene complex to the sex discrimination of cancer risk in 19 human neoplasias. The results obtained are as follows: a) the correlation coefficient r seq of the sequential regression analysis, as applied to 47 log AAIR data sets of one tumor pair, served as a criterion in testing the balance of power between oncogene activation and tumor suppressor gene inactivation. Sole activation of the oncogene should give an r seq value of -1.0, whereas sole inactivation of the tumor suppressor gene should give an r seq value of +1.0. b) Esophageal cancer and laryngeal cancer, two sex-discriminating tumors with distinct male predominance were each associated with differential implications of the oncogene-tumor suppressor gene complex between the male and female populations: in both tumors, the male populations were associated with a complex of activated oncogene and inactivated tumor suppressor gene, whereas the female population was associated with another complex of weakly activated (esophageal cancer) or non-activated (laryngeal cancer) oncogene and inactivated tumor suppressor gene, as assessed by the r seq criteria. c) The intersex correlation of cancer risk in both esophageal cancer and laryngeal cancer for 47 populations throughout the world, was rather weak, when compared with other members of human neoplasias. The intersex difference of r seq as expressed in terms of t value of Student's t test for each of 19 human neoplasias, was negatively correlated with the correlation coefficient r of the intersex regression analysis with the same 47 populations. It was indicated that a change in the intersex linkage of cancer risk may be related to the differential implication of the oncogene-tumor suppressor gene complex in carcinogenesis. In summary, we conclude that the hormonal milieu of the host plays a cardinal role as the modifier of the oncogene tumor suppressor gene impact. PMID- 9216704 TI - Outcome of patients with lung cancer detected by mass screening versus presentation with symptoms. AB - Lung cancers in the early stages are frequently detected via mass screening in Japan. The aim of this study is to evaluate the outcome of patients with lung cancer detected via mass screening and to compare them to those in whom the malignancy was detected by symptoms. A total of 774 untreated patients with lung cancer who were admitted to Department of Respiratory Medicine, Tsukuba University Hospital over a 20 year period up to 1995, were analyzed with reference to their reasons for detection of the cancer. In the mass screened group(116 patients), 50.0% of lung cancer was detected at stage I of TNM classification, while only 8.2% of patients with symptoms(561 patients) had stage I lung cancer (p = 0.0001). As lung cancers detected via mass screening were more often at operable stage (stage I, II or IIIA) (p = 0.0001), surgical treatment was chosen more frequently in the mass screened group(p = 0.0001). The outcome of patients with lung cancer detected via mass screening was more favorable than that of the patients detected by their symptoms (p = 0.0002). The early detection of lung cancer via mass screening contributes to improvement of the outcome. PMID- 9216705 TI - Endometrial carcinoma: results of primary surgery on FIGO stages Ia-Ic and predictive value of histopathological parameters. AB - The aim of the present investigation was to see if alternative histopathological parameters could identify a smaller high risk group than commonly seen using routine histopathological parameters. The material consisted of 150 primary resected patients of FIGO Ia-Ic diagnosed as endometrial carcinoma and 12 cases of atypical hyperplasias which were suspected to contain small areas of carcinoma. The patients were treated from December 1979 to April 1993 at the Department of Obstetrics and Gynecology, University Hospital, Lund, Sweden. Those with deep myometrial invasion (> 50%) were given external radiotherapy (20-30 Gy) postoperatively. The follow-up period ranged from 2.5 to 5 years with 116 patients followed-up for more than 5 years. As no therapy was given before surgery we could investigate histopathologic variables such as degree of differentiation and cytology, number of mitoses per high power field (x 40), nuclear polymorphism, mode of invasion, the extension of myometrial invasion, vessel invasion as well as grade of lymphocyte reaction around the tumour cells. We found the degree of differentiation, vessel invasion, number of mitoses, mode of invasion and cytologic abberation to be significant prognostic parameters. The frequency of deep myometrial invasion (> 50%) was extremly high (51/150 = 33%). However, this usually strong parameter was only significant when comparing stage Ia with Ic. Thus the prognostic capacity of myometrial invasion is diminished in primary hysterectomized patients. In the regression analyses only vascular invasion remained significant. By combining vascular invasion with the degree of differentiation we diminished the high-risk group consisting of candidates for further investigation and treatment. Thus a high risk group consisting of poorly differentiated carcinomas with vascular invasion was constructed comprising 24 of 139 patients with a mortality rate of 60%. PMID- 9216706 TI - Supraclavicular lymph node metastases (SLM) from breast cancer as only site of distant disease: has radiotherapy any role? AB - BACKGROUND: Supraclavicular lymph node metastases (SLM) as the only site of metastatic disease from breast cancer is a rare and a poor prognostic event. In order to evaluate the role of Radiotherapy (RT) with "radical dose" to the supraclavicular fossa, we carried out a non randomized clinical trial comparing systemic therapy alone to integrated and aggressive treatment (systemic therapy plus radiotherapy). The primary end-point was time to progression (TTP). The second end-point was the overall survival (OS). METHODS: From 1/1/1989 to 31/12/1994 37 patients (with or without the presence of locoregional disease) were enrolled into two arms, of the study, but were allowed, when giving their consent, to change the arm of the study which they had been originally allotted to. Arm A, 18 patients, 15 evaluable: chemo +/- hormonotherapy for 6 courses; after the second course, if local progression disease was present, the pts. were submitted to RT and removed from the study (3 patients). Arm B, 19 patients all evaluable: chemo +/- hormonotherapy for 3 courses followed by RT with "radical" dose. Results were analyzed on 30/11/1995 and no interim analysis was performed. The potential median follow up for all patients was 56.5 months (range 11-83 months): for Arm A 61 months (range: 12-82); for Arm B 53 months (range: 11-83). The two groups were homogeneous and balanced, without statistical differences. RESULTS: Median TTP was 12.5 months in Arm A and 19.5 months in Arm B (p = 0.064). Median overall survival (OS) was 27.5 months in Arm A and 48 months in Arm B. T-status to the time of the diagnosis was found to be independent prognostic factor for TTP (p = 0.0029). Disease-free interval from diagnosis to recurrence was found to be a significant prognostic factor for OS (p = 0.009). CONCLUSION: The results in Arm B demonstrated the opportunity of a long term control in this subset of patients. Therefore we suggest the start of a wider multicenter study in order to define the biological significance of SLM, its importance in staging breast cancer and to consider the optimum treatment. PMID- 9216707 TI - Serum tissue polypeptide specific antigen (TPS) in patients with cervical carcinoma: preliminary report. AB - Tissue polypeptide specific antigen (TPS) was measured by TPS ELISA in the sera of 88 patients with FIGO stage II and III cervical cancer and 93 healthy Thai women as the control group. The mean serum TPS levels were 63.1 U/L in the control group, and 166.4 and 363.2 U/L in stage II and III cervical cancer respectively. The mean of the control group and stage II patients were not significantly different while the mean of stage III patients was significantly different from those two groups (p < 0.0005). With the cut-off value of 90 U/L, the rates of TPS elevation were 22/35 (65.7%) in stage II and 42/53 (79.2%) in stage III patients. As for the pathology, squamous cell carcinoma showed a statistical difference from adenocarcinoma and adenosquamous carcinoma of P = 0.001. For squamous cell carcinoma, there was no difference between the keratinized and non-keratinized type (P = 0.451). TPS is not sensitive in stage II. However, it might be useful for predicting prognosis if the elevation is significantly high, and distant metastases or local recurrence should be investigated. PMID- 9216708 TI - Effects of 5'-DFUR and OK-432 on cytokines and thymidine phosphorylase in tumor tissue of gastric cancer patients. AB - BACKGROUND: The purpose of the study was to verify whether OK-432 in combination with 5'-DFUR induced thymidine phosphorylase (TdR Pase) and cytokines in gastric cancer patients as well as in vitro. MATERIALS AND METHODS: Fifty patients with invasive gastric cancer were randomly assigned, upon admission using by a closed envelope technique, to either a group receiving 5'DFUR or OK-432 alone, to a group receiving both 5'DFUR and OK-432, or to a non- treated group up. Surgical specimens of the tumor and normal tissues were taken soon after gastrectomy to evaluate TdR Pase activity, IL-1 alpha and TNF-production. RESULTS: TdR Pase activities were several times higher in tumor than in normal tissues. In normal tissues, TdR Pase activities in the 5'-DFUR + OK-432 group were significantly higher than in the OK-432 group. TdR Pase activity in tumors, however, showed no significant difference between treated group. In the 5'-DFUR + OK-432 group, the level of IL-1 alpha production in tumor was significantly higher compared to the control group. In the 5'-DFUR + OK-432 group, the level of TNF alpha production in tumor was significantly higher than in normal tissue. TNF alpha production in tumor showed no significant difference in each treated group compared to the control. There was a significant correlation between TdR Pase activity and IL-1 alpha production levels in tumor. CONCLUSIONS: TdR Pase was induced by IL-1 alpha in tumor tissues of gastric cancer patients. OK-432 in combination with 5'-DFUR, however, did not induce TNF alpha and IL-1 alpha, and increase TdR Pase activity in gastric cancer tumors. PMID- 9216709 TI - Thrombomodulin is a new biological and prognostic marker for breast cancer: an immunohistochemical study. AB - BACKGROUND: Thrombomodulin (TM) is a natural anticoagulant which inhibits thrombin. Recent studies have reported that TM is correlated with vascular diseases and a few cancers. The aim of this study was to evaluate the role and the prognostic value of TM in breast cancer. PATIENTS AND METHODS: TM expression in samples from 60 invasive breast cancer patients was examined immunohistochemically with a polyclonal antibody against TM. RESULTS: TM staining was observed mainly on both the cytoplasm and cell surface in cancer cells and on endothelial cells around or in cancer tissue. TM expression in cancer cells was not correlated with the clinicopathological features. However, low TM expression was significantly correlated with a high relapse rate (p = 0.047 by the chi 2 test and 0.05 by the Kaplan-Meier method). CONCLUSIONS: TM might play an active role in cancer invasion and metastasis, and serve as a new prognostic factor in invasive breast cancer. PMID- 9216710 TI - Intensified M-VEC chemotherapy with G-CSF support as outpatient treatment for advanced bladder cancer. AB - The M-VAC regimen (methotrexate, vinblastine, doxorubicin and cisplatin) is widely used in the treatment of advanced or metastatic bladder cancer. In the present trial, an alternate week regimen of M-VEC (with epidoxorubicin instead of doxorubicin) supported by G-CSF (filgrastim) was evaluated. Eligible patients had metastatic or surgically unresectable bladder cancer, not previously treated with systemic chemotherapy. Treatment consisted of methotrexate 30 mg/m2 day 1, vinblastine 3 mg/m2 day 2, epidoxorubicin 60 mg/m2 day 2 and cisplatin 35 mg/m2 days 2 and 3. G-CSF was administered s.c. at the dose of 300 mcg from day 4 to day 11: the treatment was repeated every 2 weeks for six cycles. Twenty-one patients entered the study, and 19 received more than 1 cycle: 78.9% were able to receive full doses of M-VEC as scheduled. The treatment resulted feasible on an outpatient basis, with mild toxicity. Three complete responses (15.8%) and 9 partial responses (47.4%), with an overall objective response rate of 63.2%, were observed. PMID- 9216711 TI - Merkel cell carcinoma of the skin. Treatment of primary, recurrent and metastatic disease: review of clinical cases. AB - The clinical features of 10 cases of primary neuroendocrine carcinoma of the skin (Merker cell tumor) are reported. This cancer arises in the dermis and subcutaneous tissues of elderly individuals. Natural history is characterized by local recurrences (30%), regional lymph node metastases (65%) and distant metastases (40%). Surgery is elective treatment, chemotherapy and radiotherapy resulted only to short-term palliative response. PMID- 9216712 TI - Immunocytochemical detection of prostate specific antigen expression in human primary and metastatic melanomas. AB - Prostate-specific antigen (PSA), a 33 kD glycoprotein, was initially reported to be a tissue specific protein, detected in the seminal fluid and produced by normal and abnormal epithelial cells of the prostate gland, as well as other tissues in the human body. The expression of PSA has been described to be elevated during benign and neoplastic cell growth in the prostate, and in a number of other human malignancies. The presence and production of PSA in human primary cutaneous malignant melanomas (CMMs) and metastatic malignant melanomas (MMMs) has not been reported prior to the present study. We examined the expression of PSA employing a biotin-streptavidin based, alkaline phosphatase conjugated antigen detection technique in routine, neutral formalin fixed, paraffin-wax embedded, 3-4 microns thick tissue sections of 30 CMMs and 10 MMMs. Human postnatal thymic tissue, among others, was used as a negative tissue control, while normal prostate and prostate carcinomas (PCs) were included in the collection of antigen positive tissues. We observed the presence of PSA in 16/30 CMMs and 6/10 MMMs. The intensity of the staining was moderate (C to B) and localized to between 20% and 30% of the total tumor cell population in both CMMs and MMMs, with cells of similar immunoreactivity being clustered in groups within the tumor microenvironment. This result directly contradicts the previous opinion concerning the prostate epithelium specificity of PSA expression and production. The immunophenotype (IP) heterogeneity of malignant melanoma cells in further substantiated by the pattern of their PSA immunoreactivity. The establishment of the clinical significance of these findings necessitates further in vivo and in vitro research in malignant melanomas. PSA related, novel antineoplastic immunotherapy may also be recommended in the treatment of both CMMs and MMMs. PMID- 9216713 TI - Fine mapping of the human renal oncocytoma-associated translocation (5;11)(q35;q13) breakpoint. AB - Recent cytogenetic analysis of a series of human renal oncocytomas revealed the presence of a recurring chromosomal translocation (5;11)(q35;q13) as sole anomaly in a subset of the tumors. The molecular characterization of this translocation was initiated using two primary t(5;11)-positive renal oncocytomas and a panel of somatic cell hybrids derived from one of these tumors, in conjunction with fluorescence in situ hybridization (FISH) and Southern blot analysis. The breakpoint in chromosome band 11q13 could be located within a genomic interval of at maximum 400 Kb immediately centromeric to the BCL1 locus. PMID- 9216714 TI - A recurrent translocation, t(16;21)(q24;q22), associated with acute myelogenous leukemia: identification by fluorescence in situ hybridization. AB - Chromosome fluorescence in situ hybridization (FISH) analyses were performed on bone marrow cells in 3 adult patients with MDS or AML with a (16;21)(q24;q22) translocation. FISH analyses with AML1 probes at 21q22 proved in all 3 patients splitting of the AML1 gene at a region spanning exons 5 and 6 and the translocation of its 5' segment to distal 16q. Chromosome painting FISH analysis in patient 1 proved the translocation of the distal 21q segment to 16q, but it failed to prove the presumed translocation of the distal 16q segment to 21q, most likely because of its small size. PMID- 9216716 TI - Translocation (8;12;21)(q22.1;q24.1;q22.1): a new masked type of t(8;21)(q22;q22) in a patient with acute myeloid leukemia. AB - The translocation t(8;21)(q22;q22) is found in 40% of cases of acute myeloid leukemia (AML) designated as the subtype M2 in the French-American-British (FAB) classification. The 8;21 translocation is clinically of interest because patients with this subtype have a good prognosis. We describe a masked type of the translocation, t(8;12;21)(q22.1;q24.1;q22.1). The translocation was first interpreted as t(8;12)(q22;q24) based on cytogenetics, but was reevaluated as a result of Southern blot and fluorescence in situ hybridization (FISH) analyses. PMID- 9216715 TI - Band 1p36 abnormalities and t(1;17) in ovarian carcinoma. AB - In a series of 128 karyotyped ovarian carcinomas, 42% of cases with chromosome 1 clonal structural abnormalities had breaks at band 1p36 (usually involving translocations of unknown material). Fluorescent in situ hybridization (FISH) studies using combinations of 1 centromere and 1p36.3-specific probes (16 cases) or 1 centromeric and 17 whole-chromosome paint probes (11 cases with 1p+) revealed a trend toward deletion of 1pter relative to 1 centromere (63%); intratumor heterogeneity; and the origin of 1p+ in 3/11 cases (27%) from chromosome 17 [t(1;17)(p36;?)]. The frequency of this specific breakpoint and its involvement in recurrent translocations suggest that these regions are loci for genes important in the pathogenesis of a subset of sporadic ovarian carcinomas. PMID- 9216717 TI - Granulocytic sarcoma with translocation (9;11)(p22;q23): two cases. AB - Granulocytic sarcomas are localized deposits of myeloid leukemia cells that may precede or occur concurrently with disseminated disease. In either event, the origins of the cells comprising the malignancy are the same. Published reports of granulocytic sarcomas have described, in the majority of cases, a morphology typical of AML-M2 and the presence of the t(8;21)(q22;q21) typical of that FAB type. In a smaller number of cases, the inv(16)(p13q22) characteristic of AML-M4 has been recorded in cells with a myelomonocytic appearance. We report two patients with granulocytic sarcomas showing monocytic morphology in which the malignant cells showed t(9;11)(p22;q23) typical of AML-M5. This abnormality is seen in up to 7% of childhood AML, but has not previously been reported in granulocytic sarcoma. The detection of this cytogenetic abnormality facilitated the precise characterization of the malignant cells and selection of the most appropriate therapy, emphasizing the value of cytogenetic analysis in cases of granulocytic sarcoma. PMID- 9216719 TI - Characterization of chromosome changes in two human prostatic carcinoma cell lines (PC-3 and DU145) using chromosome painting and comparative genomic hybridization. AB - Using chromosome painting, a study of chromosomal abnormalities has been performed in two prostatic carcinoma cell lines, PC-3 and DU145. In PC-3, this analysis revealed a highly rearranged hypotriploid karyotype with 54 to 61 chromosomes and numerous rearrangements of chromosomes 1, 3, 5, 8, 10, and 14. At passage 73, DU145 had a hypotriploid karyotype with few rearrangements of chromosomes 1, 3, 5, 12, 13, and 20, whereas at passage 153, this cell line showed a near-tetraploid karyotype with a great number of rearrangements involving chromosomes 3, 6, 8, 10, 12, and 17. A single rearrangement was shared by the 2 cell lines, an i(5)(p10). A comparative genomic hybridization study demonstrated a noticeable amplification of bands 10q22.3-q23 and 14q22-q24 in the PC-3 cell line. No amplification signal was detected for DU145. PMID- 9216718 TI - Diffuse large cell, B-cell type lymphoma with a novel translocation (2;22)(p23;q11.2). AB - We observed a translocation (2;22)(p23;q11.2) in the bone marrow cells of a patient with multiple subcutaneous nodules. Tumor histology and immunohistochemical staining demonstrated a malignant lymphoma, diffuse large cell type, displaying a CD30 negative B cell immunophenotype. To our knowledge, this is the first report of this specific translocation in lymphoma, which may join the site of the anaplastic lymphoma kinase (ALK) gene at 2p23 to the region of the immunoglobulin lambda light chain gene at 22q11.2. The ALK gene was initially identified through its involvement in the t(2;5)(p23;q35) found most commonly in anaplastic large cell lymphoma. This observation in a CD30 negative large cell lymphoma of B cell lineage further extends the relationship of anaplastic large cell morphology, ALK activation, lymphoid lineage, and expression of the CD30 antigen. PMID- 9216720 TI - HMGIC expressed in a uterine leiomyoma with a deletion of the long arm of chromosome 7 along with a 12q14-15 rearrangement but not in tumors showing del(7) as the sole cytogenetic abnormality. AB - Cytogenetic studies on uterine leiomyomas have shown that more than 60% of these tumors possess a normal karyotype and that 30% have clonal chromosomal aberrations. The most frequent changes are aberrations involving 12q14-15 and show rearrangements of the long arm of chromosome 7. Recently, we were able to demonstrate that in a variety of mesenchymal tumors showing 12q14-15 aberrations the HMGIC gene is rearranged thus playing a role in tumorigenesis. Here we report the results of HMGIC expression studies by RT-PCR of five uterine leiomyomas with different karyotypes. The RT-PCR studies were performed on two primary tumors showing a 12q14-15 aberration, one of which with an additional del(7) and three tumors with del(7) as the sole aberration. The tumor with the 12q14-15 aberration as the sole alteration and the leiomyoma with 12q14-15 rearrangement plus deletion of the long arm of chromosome 7 were shown to express HMGIC. In contrast, in all three tumors with the del(7) as the sole aberration no expression of HMGIC was noted. PMID- 9216721 TI - Loss of heterozygosity at chromosome 9p21 in primary neuroblastomas: evidence for two deleted regions. AB - The genes responsible for the development of neuroblastoma following in vivo deletion or mutation are largely unknown. We have performed loss of heterozygosity studies on a series of 24 Portuguese primary neuroblastomas using 6 polymorphic markers located at chromosome 9p21 spanning the p16/MTS1/CDKN2, p15/MTS2/CDKN2B, and the interferon alpha and beta genes. Loss of heterozygosity was observed in 4 of the 24 tumors (17%), a somewhat lower percentage than a previous study that identified patients by a mass screening program. A correlation was also observed between 9p21 LOH and 1p36 LOH in our group of tumors. Two distinct regions of 9p21 deletion were observed: one located in the region adjacent to the markers D9S162 and D9S1747 and a second located centromerically of the p16 gene near the D9S171 marker. The latter region is exclusive of the p16 gene. This result suggests the presence of at least one other tumor suppressor gene at 9p21, apart from the p16 and p15 genes, which may be of importance to the development of neuroblastoma. PMID- 9216722 TI - Aneuploidy and consistent structural chromosome changes associated with transformation of Syrian hamster embryo cells. AB - To gain a better understanding of the role of specific numerical and structural chromosome changes in the multistage process of transformation of Syrian hamster embryo (SHE) cells, we analyzed seven benzo(a)pyrene (BP)-induced immortal SHE cell lines, and one spontaneously immortalized cell line. In addition, we analyzed chromosome changes in early passage tumor-derived cell lines induced by injection of four immortalized cell lines into neonate hamsters. Of particular interest was the observation of a deletion in the short arm of chromosome 2 in four of the seven BP-immortalized cell lines. Other types of alterations in chromosome 2 were observed in two other cell lines. Loss of one copy of chromosome 16 was also observed in more than 90 to 100% of the cells in three of seven BP-immortalized cell lines. In contrast, the only chromosome alteration seen in the spontaneously immortalized cell line was a deletion in the short arm of chromosome 20. Genetic instability, as indicated by increased numerical or structural chromosome changes, was observed in all tumor-derived cell lines compared to the immortal cell line from which they originated. These results, along with previous reports in the literature, suggest that alterations in specific chromosomes, like chromosome 2, may be involved in transformation of SHE cells. PMID- 9216723 TI - Routine karyotyping in Wilms tumor. AB - We describe the karyotypes of nine Wilms tumors (WT). Four tumors were initially karyotyped from diagnostic needle core biopsies, 3 after postchemotherapy tumor resection and the remainder from xenografts grown in nude mice. The 9 nephroblastomas were composed of 7 with favorable histology (intermediate-grade malignancy) and 2 with unfavorable histology (anaplastic or high-grade malignancy). The 7 tumors with favorable histology had karyotypes typical of WT, with the previously described nonrandom abnormalities +1q, +6, +7, +8, +12, +13, +18 and structural abnormalities of 1p and 16q present in at least 1 case. The most common abnormalities were trisomy 18 (4 cases) and +1q (3 cases). The 2 tumors with unfavorable histology both had complex karyotypes atypical for WT. We suggest that cytogenetics can act as a marker when histologic grade is in doubt. Karyotypic analysis from needle core biopsies was attempted in 6 samples, including 1 from a nephrogenic rest (NR) of the nonaffected kidney and provided a result on 5 occasions. The NR were present in the sole case with a constitutional abnormality, a mosaic partial duplication of 8q. However, both the tumor and the NR were apparently derived from the normal cell line. Here we demonstrate that a cytogenetic result can be routinely obtained from needle core biopsies and will thus facilitate true diagnostic tumor karyotypes in both WT and other tumors. PMID- 9216724 TI - Increased frequency of P-glycoprotein gene amplification in colchicine-resistant Rat-1 clones transformed by v-src. AB - A rat fibroblast cell line (Rat-1) carrying a temperature-sensitive mutation of v src was used to determine whether inducible cellular transformation altered the ability of cells to amplify the p-glycoprotein gene in response to colchicine selection. Transformed and nontransformed Rat-1 fibroblasts selected under 4 times the LD50 generated the same number of colchicine-resistant colonies. We next examined colchicine-resistant colonies derived from transformed cells and compared them to colchicine-resistant colonies derived from nontransformed cells. When Rat-1 cells were selected at 35 degrees C (transforming temperature), 7 out of 7 clones exhibited a 3- to 5-fold p-glycoprotein gene amplification. These results contrasted to those found at the nontransforming temperature (40 degrees C); none of the 8 colchicine-resistant clones examined had amplified the p glycoprotein gene. Thus in Rat-1 cells carrying a temperature-sensitive v-src gene, p-glycoprotein gene amplification was observed at a high frequency only in transformed clones selected at the temperature permissive for v-src activity. PMID- 9216725 TI - Karyotypic analysis of 32 malignant epithelial ovarian tumors. AB - The identification of recurrent specific cytogenetic findings in various malignancies has provided an improved means to diagnose and treat patients. To date, no characteristic markers have been found for epithelial ovarian cancer. This is due, in part, to several contributory factors, including the inability to identify optimal growth conditions for culture and the fact that most analyses of advanced-stage tumors are obtained from malignant effusions rather than from solid tissue. In addition, many reports include previously treated patients. In this study, 32 untreated solid epithelial ovarian tumors, including 8 tumors of low malignant potential (LMP), were obtained from primary and metastatic sites at initial surgical staging. Using a 2-culture plastic technique for tissue growth, we achieved a 96% short-term culture success rate. Only 4 normal 46,XX karyotypes were identified. Diploid or near-diploid genomes were associated with few cytogenetic alterations. Complex karyotypic morphologies were consistently associated with advanced or poorly differentiated tumors. Nonrandom cytogenetic aberrations most commonly involved chromosomes 1 and 6. A novel translocation, t(1;6)(p10;p10), was identified in both a metastatic LMP tumor and a poorly differentiated invasive tumor. This cytogenetic rearrangement can potentially be regarded as a clinically relevant early marker for tumorogenesis. Finally, karyotypes from both primary and metastatic sites were subject to a comparative analysis in 11 patients. In 4 cases, greater chromosomal complexity was associated with the primary site. PMID- 9216726 TI - A Ph-negative chronic myeloid leukemia patient with a non-classical BCR-ABL rearrangement characterized by fluorescence in situ hybridization. AB - A patient with chronic myeloid leukemia (CML), a normal karyotype and a BCR-ABL rearrangement is presented. Southern blot analysis detected the rearrangement, whereas RT-PCR with b2a2 and b3a2 primers did not. Fluorescence in situ hybridization (FISH) with an ABL probe (9q34.2) and an Mbcr probe (22q11) showed ABL and BCR signals on chromosome 22. Subsequent FISH studies with cosmids mapping to 9q34.3 showed normal hybridization patterns to chromosome 9, suggesting an interstitial insertion of ABL containing DNA sequences into chromosome 22 in this patient. The lack of reciprocal translocation sequences was investigated with RT-PCR, primers a1b and c7. The absence of ABL-BCR gene expression in this and other patients described in the literature with this subtype of Ph-negative CML, does not seem to have an impact on the clinical course of the disease. PMID- 9216727 TI - t(5;7)(q34;q21) in acute megakaryoblastic leukemia associated with Down syndrome. PMID- 9216728 TI - Involvement of 8q, 22q, and monosomy 21 in an epithelioid sarcoma. PMID- 9216729 TI - Addition (1)(q32) as the sole clonal chromosomal abnormality in a case of cardiac myxoma. PMID- 9216730 TI - Cytogenetic findings in a bladder chondrosarcoma. PMID- 9216731 TI - Cellular localization of thrombopoietin mRNA in the liver by in situ hybridization. AB - The expression of thrombopoietin (TPO) mRNA is observed in several tissues, including liver, kidney, brain, skeletal muscle, intestine, spleen, and bone marrow. Among these organs, the highest expression of TPO mRNA is detected in the liver. We identified cells producing TPO by means of in situ hybridization of adult rat liver using digoxigenin-11-UTP-labeled cRNA probes. We found that the cells expressing TPO mRNA also expressed serum albumin mRNA. TPO mRNA was detected in parenchymal cells (hepatocytes) but not in non-parenchymal cells (including endothelial cells, epithelial cells, and so forth). To determine the location of TPO expression in embryogenesis, sections of fetal mice were further analyzed by in situ hybridization. TPO mRNA was detected only in hepatocytes of fetal liver, which was also the major site of hematopoiesis. The expression of TPO mRNA in fetal liver was observed from 12.5 days postcoitus. Northern blot analysis showed that mouse liver transcribed the same size of TPO mRNA in the fetus and in the adult. These results clearly demonstrate that hepatocytes are the primary site of TPO production in the liver from fetus to adult. PMID- 9216733 TI - Nontransformed colony-derived stromal cell lines from normal human marrows. III. The maintenance of hematopoiesis from CD34+ cell populations. AB - Nontransformed stromal colony-derived cell lines (CDCLs) consist of a pure stromal cell population that differentiates following a vascular smooth-muscle cell repertoire. Here we study the maintenance of hematopoiesis by this cell population. We show that CDCLs allow the generation for several weeks of stroma adherent colonies (comprising a cobblestone area) from CD34+, CD34+/CD38+, and CD34+/CD38- cells. Stroma-adherent colony-forming cells (CFCs) from CD34+/CD38- cells reach a maximum at week 4 and limiting dilution analysis gives a frequency of 1 per 10 cells seeded; in contrast to this, CFCs from CD34+/CD38+ cells are optimal by week 2 and the frequency is then only 1 per 120 cells seeded. Stroma adherent colonies comprise hematopoietic cells from all lineages except the T lymphocytic, with a majority of granulomonocytes. CDCLs also allow the amplification of granulomonocytic colony-forming units (CFU-GMs), since cumulative outputs of CFU-GMs by week 6 are 190 and 8 times that observed at culture inception for the CD34+/CD38- and CD34+/CD38+ cell populations, respectively. Our results suggest that stromal cells from CDCLs allow the maintenance of primitive hematopoietic precursors and induce their proliferation and differentiation. This study underscores the potential role of one of the microenvironmental cell populations, that of myoid cells, in the regulation of hematopoietic precursor behavior. PMID- 9216732 TI - Effect of flt3 ligand on in vitro growth and expansion of colony-forming bone marrow cells from patients with aplastic anemia. AB - To determine the value of flt3 ligand (flt3L) in stimulating hematopoiesis in human hypoproliferative bone marrow disorders, we examined its in vitro effect on bone marrow cells from patients with aplastic anemia (AA). Growth response to flt3L, alone and in combination with other hematopoietic growth factors, was investigated in clonogenic methylcellulose assays, in long-term liquid and stroma cultures. Bone marrow cells were derived from 13 AA patients with persisting in vitro growth defect after immunosuppressive treatment and from nine normal bone marrow donors. In methylcellulose cultures, flt3L stimulated formation of hematopoietic colonies only weakly, whereas it had an additive effect when combined with erythropoietin (Epo), stem cell factor (SCF), interleukin-3 (IL-3), interleukin-11 (IL-11), or granulocyte colony-stimulating factor (G-CSF). Flt3L was less effective than SCF and did not further enhance the number of hematopoietic colonies formed in response to SCF-containing combinations of multiple cytokines. In long-term liquid suspension cultures, flt3L was less mitogenic than SCF but its effect on the maintenance of progenitors was superior that of SCF and of IL-3, IL-11, and G-CSF. The total number of clonogenic AA cells increased as much as four-fold during the first culture week and FACS analysis demonstrated expansion of the CD34+CD38+ progenitor cell subset. Despite this enhancement, survival of AA cells remained significantly poorer than that of normal cells, in which the primitive subset of CD34+CD38- cells was maintained up to 4 weeks when flt3L was used as a single factor. Both in normal and AA cultures, flt3L promoted differentiation of cells of the myeloid lineages. In cultures of bone marrow stroma, flt3L had almost no effect on growth and survival of AA progenitors, while in cultures of normal cells the number of colony-forming cells increased up to 10-fold. Although flt3L does not overcome the proliferative defect of AA precursors, we conclude that the ligand is capable of in vitro stimulation and expansion of the reduced progenitor cell pool in AA, when used in appropriate culture conditions. The in vitro effects of flt3L on AA cells differ in many aspects from those of the structurally related cytokine SCF, suggesting a benefit in use of a combination of these two early-acting growth factors. PMID- 9216734 TI - Macrophages from motheaten and viable motheaten mutant mice show increased proliferative responses to GM-CSF: detection of potential HCP substrates in GM CSF signal transduction. AB - Loss of functional hematopoietic cell phosphatase (HCP) underlies severe hematopoietic and immunologic abnormalities in mice homozygous for the motheaten and viable motheaten mutations. These mice die from pulmonary accumulation of macrophages that are regulated by macrophage colony-stimulating factor (M-CSF) and granulocyte (G)-M-CSF. We determined the growth response of motheaten macrophages to the two growth factors and looked for potential HCP substrates in these cells. Motheaten macrophages showed increased proliferative responses to GM CSF but not to M-CSF, demonstrating that HCP plays a critical role in downregulating GM-CSF mitogenic signaling. Despite the heightened growth responses of the motheaten macrophages to GM-CSF, there were no marked differences between motheaten macrophages and normal controls in GM-CSF-induced phosphorylation of GM-CSFR beta, Jak2, STAT5 and MAPK, indicating that these molecules are not major HCP substrates in GM-CSF signaling. Interestingly, several markedly hyperphosphorylated proteins were detected in the motheaten macrophages, including a novel 126-kDa phosphotyrosine protein that associated with the phosphatase via its SH2 domains, suggesting that these proteins depend on HCP for dephosphorylation and may mediate the heightened growth responses to GM-CSF. Our data indicate that macrophage hypersensitivity to GM-CSF may be a major factor in motheaten pathogenesis and that HCP may dephosphorylate novel substrates critical in GM-CSF mitogenic signaling. PMID- 9216735 TI - Membrane-bound proteindisulfide isomerase (PDI) is involved in regulation of surface expression of thiols and drug sensitivity of B-CLL cells. AB - The proteindisulfide isomerase (PDI), a multifunctional cytoplasmic enzyme with additional chaperone activity, has been shown recently, using monoclonal antibodies, to be located on the membrane of mature human B lymphocytes and B cell chronic lymphocytic leukemia (B-CLL) cells. Here, evidence is presented that this antigen exhibits catalytic activity as measured by the reductive degradation of insulin (release of A chain molecules) on intact B cells in patients suffering from B-CLL, as well as on JVM 13 cells (B-CLL cell line). More than 98% of these cells exhibited PDI activity which could be inhibited by bacitracin and also by monoclonal and polyclonal antibodies to PDI. Interestingly, surface PDI expression was strongly correlated in our study with protein-bound membrane SH groups. These surface protein thiols were specifically determined by using low concentrations of the chloromethyl-derivative based fluorescent probe 5-(and6) (((4-chloromethyl)-benzoyl)amino)-tetramethyl-rhodamine (CMTMR) at low temperature in the presence of sodium azide in flow cytometry. The highest PDI and SH expression was found on B lymphocytes, particularly B-CLL cells. The mean fluorescence intensity (MFI) of CMTMR-positive B cells in the B-CLL line was up to 10-fold higher than that of controls, indicating a strong elevation of cell membrane-located protein thiols on malignant B cells. The link between PDI and SH expression on cell surfaces points to a functional interaction between the two. Treatment with bacitracin resulted in a strong inhibition of PDI and a dramatic increase in surface protein thiol expression of B-CLL cells. Similar effects could be observed by cell treatment with anti-PDI antibodies, indicating that this enzyme system plays a crucial role in the regulation of protein-bound SH groups. Interestingly, artificially induced protein thiol expression led to significantly higher cellular resistance to the cytostatic drugs chlorambucil, vinblastin, and cisplatin in vitro as measured by cell growth. These data suggest for the first time a regulatory effect of PDI on the surface protein thiol status of B cells. The increased expression of PDI may play a crucial role in SH mediated protection and drug resistance in malignant B lymphocytes. PMID- 9216736 TI - Expression and function of the peptide transporters in escape variants of human renal cell carcinomas. AB - The transporter associated with antigen processing (TAP) complex is a heterodimeric transmembrane pump consisting of the TAP-1 and TAP-2 subunits; these subunits translocate peptides from the cytoplasm into the lumen of the endoplasmic reticulum, where they bind nascent major histocompatibility complex (MHC) class I molecules. Loss or reduced expression of the TAP genes results in the synthesis of unstable peptide free MHC class I molecules that are only weakly expressed on the cell surface. In a number of human tumor cell lines, downregulation of MHC class I expression has been found to be associated with reduced or absent TAP expression. To investigate whether alterations in MHC class I expression occur during transformation and subsequent progression and whether MHC class I suppression is caused by impaired TAP function, we analyzed the protein expression of MHC class I heavy and light chain and TAP-1 in three renal cell carcinoma (RCC) cell lines and short-term cultures from corresponding normal kidney tissue. In one case a cell line established from a metastatic lesion was also available. Compared with normal epithelial cells, suppression of TAP-1 and MHC class I molecules was detected in all three primary RCC cells and was even more pronounced in the metastatic cell line. In contrast to normal epithelial cells, MHC class I membrane expression of two RCC lines was enhanced by culture in the presence of MHC class I binding peptides or at low temperature (26 degrees C) instead of 37 degrees C. Unstable MHC class I surface expression is caused by dissociation of the MHC class I heavy and light chain molecules as a result of functional defects in the antigen processing machinery, e.g., impaired peptide transport. Attempts to counteract the reduced immunogenicity by transferring the TAP genes into these cells demonstrated that TAP-1-modified RCC cells expressed higher levels of MHC class I molecules. These data indicate that downregulation and instability of MHC class I surface expression in RCC cells is at least partially caused by deficient loading with endogenous peptides and can be restored by TAP gene transfer. PMID- 9216737 TI - Pathophysiology of bleeding in heat stress: an experimental study in sheep. AB - Widespread hemorrhagic manifestations commonly occur in patients with severe heat stroke. The pathogenesis of hemostatic disorders in these patients is not fully understood, although it is believed to be multifactorial in origin. The present investigation was designed to study the changes in blood platelets caused by heat stress in an experimental model of five merino sheep. The experiments were performed in two groups of five merino sheep each. In one group the sheep were subjected to a combination of heat (elevated environmental temperature) and exertional stress, and allowed to proceed throughout the experiment until a state of near collapse was reached (Task A). In the other group (Task B) the animals were heated in the same manner as those in Task A and also subjected to exertional heat; however, when the temperature reached 43.6 +/- 0.2 degrees C, the critical core temperature (CCT), they were subjected to evaporative cooling in a climatic chamber. Serial changes in the platelet counts and platelet functions were measured throughout the duration of the experiments. At the core temperature (CT) of 42.1 degrees C and above there was a significant impairment of adhesion of platelets to glass beads. During the early phases of elevation of CT, platelets showed hyperaggregation in the presence of different agonists (such as, collagen, ADP, ristocetin); this was followed by hypoaggregation when the CCT was raised above 43.6 +/- 0.2 degrees C. However, these impairments of platelet functions occurring at elevated CT and CCT were found to reverse to normal within 24 hours after the animals were cooled to 39 degrees C. It was also found that the hyperaggregation of platelets to different agonists induced by raised CT could be partially prevented by prior in vitro treatment of platelets with apyrase, a known enzyme destroying of ADP. The results of these experiments indicate that heat stress induced by exposing merino sheep to elevated controlled temperature directly activates the platelets. This may be an important contributing factor in causing altered hemostasis in heat stroke activated directly by heat. This mechanism may be operating in altered hemostasis in heat stroke. PMID- 9216738 TI - Loss of flk-2/flt3 expression during commitment of multipotent mouse hematopoietic progenitor cells to the mast cell lineage. AB - The expression of c-kit and flk-2/flt3 was analyzed in various stages of mast cell differentiation using reverse transcriptase polymerase chain reaction (RT PCR). Mouse fetal liver cells were sorted using antibodies for Sca-1 (Ly6A/E) and CD43 to obtain a population enriched for early progenitors; committed mast cell progenitors were absent from this population. Mouse fetal liver-derived, IL-3 dependent blast cell colonies provided a source of committed mast cell progenitors, and mast cell colonies provided mature mast cells. Comparison of these populations showed that some uncommitted cells express both c-kit and flk 2/flt3. At the time of commitment to the mast cell lineage, the expression of c kit increases compared to that of uncommitted progenitors, and the expression of flk-2/flt3 becomes undetectable. Previous studies have shown that steel factor, the ligand for c-kit, supports mast cell differentiation in vivo and in vitro. In contrast, the ligand for flk-2/flt3 is inactive on mast cells. Thus, receptor gene expression appears to be an important determinant of the response or lack of response of mast cells to the ligands for these two homologous receptors. PMID- 9216739 TI - Role of cytoprotective mechanisms in the photochemical purging of autologous bone marrow grafts. AB - The molecular basis of the differential sensitivity of normal hematopoietic stem cells and of leukemia, lymphoma, and neuroblastoma cells to merocyanine 540 (MC540)-mediated photodynamic therapy (PDT) is not yet completely understood. While the capacity to bind dye molecules appears to be the major determinant of a cell's susceptibility of MC540-mediated PDT, we here present evidence that under certain experimental conditions a cell's capacity to repair MC540-mediated photodynamic damage is also an important factor. Two parameters, temperature and intracellular glutathione (GSH) content, were varied to investigate the role of cellular defense mechanisms in the dye-sensitized photoinactivation of normal murine granulocyte/macrophage progenitors (CFU-GM) and K562, L1210, and melphalan resistant L1210/L-PAM1 leukemia cells. When exposed to MC540 and light at room temperature, the three leukemia cell lines bound similar amounts of dye and accumulated similar amounts of lipid hydroperoxide (LOOH) but differed markedly in their sensitivity to MC540-mediated PDT. Performing MC540-mediated PDT at 4 degrees C instead of at room temperature reduced dye binding and LOOH generation and enhanced cytotoxicity in some but not all cell lines. A brief (< or = 120 minutes) incubation at 37 degrees C immediately following MC540-mediated PDT accelerated the decay of LOOH in all leukemic cell lines and reduced cell kill by about 2 log in both CFU-GM and leukemia cells. The effect of post-PDT incubation at 37 degrees C on LOOH decay was most pronounced in K562 and least pronounced in L1210/L-PAM1 cells, whereas its effect on cell survival was less pronounced in L1210 cells than in the remaining cell types. L1210/L-PAM1 cells whose GSH content had been reduced from 8.2 to 1.6 micrograms/mg protein by incubation with buthionine sulfoximine recovered from potentially lethal photodynamic damage as rapidly as untreated L1210/L-PAM1 cells and more rapidly than wild-type L1210 cells with a GSH content of 4.5 micrograms/mg protein. Thus, with regard to capacity of L1210/L-PAM1 cells to recover from photodynamic damage, the cells' enhanced capacity to synthesize GSH appeared more decisive than intracellular GSH levels per se. Taken together, these data suggest that temperature-dependent cellular defense mechanisms are significant determinants of a cell's susceptibility to MC540-mediated PDT. The data emphasize the need for temperature control during and immediately after the photochemical purging of autologous bone marrow or peripheral blood stem cells. PMID- 9216740 TI - Localization of megakaryocytes in normal mice and following administration of platelet antiserum, 5-fluorouracil, or radiostrontium: evidence for the site of platelet production. AB - The relative contributions of various organs to platelet production is controversial. In this study, serial histologic sections of bone marrow, spleen, liver, and lung from normal C57BL/6J mice and mice that had received three different agents which perturb normal murine thrombopoiesis (platelet antiserum, 5-fluorouracil, and radioactive strontium) were examined for the presence of megakaryocytes, utilizing morphologic and immunohistochemical techniques for their identification. In liver and lung tissue, megakaryocytes (including their naked nuclei or large cytoplasmic fragments) were rare in whole cross-sections (which included blood vessels) from normal and perturbed mice, even during periods of strong stimulation of thrombopoiesis. In contrast, megakaryocyte numbers were greatly increased in bone marrow and/or spleen tissue in these circumstances. We conclude that: 1) the bone marrow and spleen are the major thrombopoietic organs in the mouse, and 2) an insignificant fraction of thrombocytopoiesis occurs in the murine liver or lung, even during periods of greatly increased platelet production or following loss of the spleen and/or bone marrow. PMID- 9216741 TI - Effects of marinating on heterocyclic amine carcinogen formation in grilled chicken. AB - This study compared heterocyclic aromatic amines in marinated and unmarinated chicken breast meat flame-broiled on a propane grill. Chicken was marinated prior to grilling and the levels of several heterocyclic amines formed during cooking were determined by solid-phase extraction and HPLC. Compared with unmarinated controls, a 92-99% decrease in 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) was observed in whole chicken breast marinated with a mixture of brown sugar, olive oil, cider vinegar, garlic, mustard, lemon juice and salt, then grilled for 10, 20, 30 or 40 min. Conversely, 2-amino-3, 8-dimethylimidazo[4,5 f]quinoxaline (MeIQx) increased over 10-fold with marinating, but only at the 30 and 40 min cooking times. Marinating reduced the total detectable heterocyclic amines from 56 to 1.7 ng/g, from 158 to 10 ng/g and from 330 to 44 ng/g for grilling times of 20, 30 and 40 min, respectively. The mutagenic activity of the sample extracts was also measured, using the Ames/Salmonella assay. Mutagenic activity was lower in marinated samples cooked for 10, 20 and 30 min, but higher in the marinated samples cooked for 40 min, compared with unmarinated controls. Although a change in free amino acids, which are heterocyclic amine precursors, might explain the decrease in PhIP and increase in MeIQx, no such change was detected. Marinating chicken in one ingredient at a time showed that sugar was involved in the increased MeIQx, but the reason for the decrease in PhIP was unclear. PhIP decreased in grilled chicken after marinating with several individual ingredients. This work shows that marinating is one method that can significantly reduce PhIP concentration in grilled chicken. PMID- 9216742 TI - Lipid peroxidation and DNA damage induced by morin and naringenin in isolated rat liver nuclei. AB - The pro-oxidant activities, as determined by lipid peroxidation and DNA strand breaks, of morin and naringenin, two polyphenolic flavonoids with molecular structures similar to quercetin, were investigated under aerobic conditions in a model system of isolated rat liver nuclei. Both flavonoids induced a concentration-dependent peroxidation of nuclear membrane lipids concurrent with DNA strand breaks. These reactions were enhanced by the metal ions iron or copper. Active oxygen scavengers catalase, superoxide dismutase and mannitol had no effect on the flavonoid-induced nuclear DNA damage in the presence of the metal ions; nuclear lipid peroxidation was partially inhibited only by mannitol. It appears that hydroxyl radicals are the initiators of the peroxidation of nuclear membrane lipids, producing peroxidation products such as peroxyl radicals that in turn may be responsible for the DNA strand breaks. Alternatively, the hydroxyl radicals produced close to the DNA backbone may induce direct site specific strand breaks that are insensitive to the free radical scavengers. These results demonstrate the pro-oxidant activities of polyphenolic flavonoids, which are generally considered as antioxidants and anticarcinogens, and suggest their possible dual role in mutagenesis and carcinogenesis. PMID- 9216743 TI - Inhibition of benzo[a]pyrene-induced mouse forestomach neoplasia and reduction of H2O2 concentration in human polymorphonuclear leucocytes by flavour components of Japanese-style fermented soy sauce. AB - Previously it was reported that 4-hydroxy-2 (or 5)-ethyl-5 (or 2)-methyl-3(2H) furanone (HEMF), a characteristic flavour component of Japanese-style fermented soy sauce that exhibits antioxidant activity, inhibits benzo[a]pyrene-induced forestomach neoplasia in mice. The antioxidant and anticarcinogenic activities of other structurally similar soy sauce flavour components are now reported. 4 Hydroxy-5-methyl-3(2H)-furanone (HMF) and 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) were found to be antioxidants. In particular, HMF and HDMF (as well as HEMF) reduced hydrogen peroxide concentration in human polymorphonuclear leucocytes stimulated by arachidonic acid or 12-O-tetradecanoylphorbol-13 acetate. HMF and HDMF were administered individually in semipurified diet to female ICR mice previously treated with benzo[a]pyrene (1.5 mg/wk, orally for 4 wk) to initiate forestomach neoplasia. The mice were killed at 30 wk of age. Both furanones reduced forestomach neoplasms, with HDMF exhibiting more potency. The data indicate that HDMF and HMF, like HEMF, inhibit carcinogenesis in this system by acting at the post-initiation stage. PMID- 9216744 TI - The effects of running stress on plasma vitamin A levels in rats. AB - The plasma concentrations of vitamin A, zinc and proteins and the hepatic level of vitamin A were determined in rats subjected to running as a model for stress and which were receiving standard or vitamin-A free diets. All rats showed a decrease in plasma vitamin A with running compared with non-running control animals. Hepatic levels of vitamin A were higher in these two test groups than in their weight- and age-matched non-running controls. The data support that running, like other forms of stress, decreases plasma vitamin A, consistent with the retention of vitamin A in the liver. PMID- 9216745 TI - Subacute and chronic oral toxicity of lornoxicam in cynomolgus monkeys. AB - Lornoxicam is a novel non-steroidal anti-inflammatory compound in the same chemical class as piroxicam and tenoxicam, with potent anti-inflammatory, antipyretic and analgesic activity. As part of the preclinical safety programme, its toxicity was evaluated in a dose-range-finding and 52-wk toxicity study in cynomolgus monkeys. In the dose-range-finding study, five groups of monkeys (two per sex per group) were dosed orally by gavage for 6 wk with 0, 0.25, 0.5, 1.0 or 2.0 mg lornoxicam/kg/day. Drug-related toxicity was observed in the 1.0 and 2.0 mg/kg/day dose groups only. This included mortality, diarrhoea, prostration, decreased body weight gain and food consumption, faecal occult blood, anaemia, leucocytosis, hypoalbuminaemia, gastrointestinal erosions and ulcerations. On the basis of these results, four groups of monkeys (six per sex per group) were given the compound orally by nasogastric intubation at dose levels of 0, 0.125, 0.25 or 0.5 mg/kg/day for 52 wk. The high-dose level was increased to 0.6 mg/kg/day from wk 39 to wk 52. Treatment was followed by a 4-wk recovery period for two animals per sex per group. Histologically, drug-related changes seen were gastrointestinal erosions, ulcerations and inflammation in males and females at 0.5/0.6 mg/kg/day. Treatment-related clinicopathological findings included decreased haematocrit and hypoproteinaemia (group 0.5/0.6 mg/kg/day males), and hypoalbuminaemia (group 0.5/0.6 mg/kg/day males and females). None of these changes were present after the recovery period, thus indicating reversibility. Plasma concentration of lornoxicam measured 2 hr after dosing increased in a dose proportional manner. The estimated area under the curve (AUC) at steady state increased in a dose-proportional manner and at 0.25 mg/kg was three- to fivefold higher than the human AUC following a 16 mg dose (8 mg b.i.d.). The no-observed effect level in the chronic toxicity study was 0.25 mg/kg/day. PMID- 9216746 TI - Lack of carcinogenicity of cyanoguanidine in F344 rats. AB - The carcinogenicity of cyanoguanidine, a compound used in the production of melamine, guanidine salts and guanamine derivatives, was examined in male and female Fischer 344 rats fed CRF-1 pulverized diets containing 0, 2.5 and 5% cyanoguanidine for up to 2 yr. The rats were randomly allocated to three groups, each consisting of 50 males and 50 females. The mean body weight gains in both sexes of the 5% group and in females of the 2.5% group were significantly lower than the control values after wk 1 of treatment. No other signs of toxicity were seen in any of the rats throughout the treatment period. Histopathologically, various tumours developed in all groups, including the control group, but these were all similar to those known to occur spontaneously in this strain of rats, and no toxicologically significant increase was found for any lesion type in the treated groups. On the basis of these results, it is concluded that cyanoguanidine exerts no carcinogenic potential in F344 rats when administered for up to 2 yr under the conditions of the present study. PMID- 9216747 TI - Effects of diethylenetriamine dihydrochloride following 13 weeks of dietary dosing in Fischer 344 rats. AB - Fischer 344 rats were fed a diet containing the dihydrochloride salt of diethylenetriamine (DETA.2HCl) at concentrations of 1000, 7500 or 15,000 ppm for 90 consecutive days. Based on food consumption and body weight, the mean corresponding dosages were 70, 530 and 1060 mg/kg/day for the males and 80, 620 and 1210 mg/kg/day for the females. Decreases in food consumption were observed intermittently throughout the dosing period in both sexes at 15,000 ppm. Dose related decreases in body weight or weight gain were observed for both sexes at 7500 and 15,000 ppm. Changes in clinical pathology measurements observed at 7500 and 15,000 ppm included increases in mean corpuscular volume and mean corpuscular haemoglobin in males, and increases in mean corpuscular volume, total leucocytes and urinary pH, and a decrease in serum glucose concentration in females. The relative kidney, brain and testes weights were increased in the 15,000 ppm males. In the females, the relative kidney, brain and liver weights were increased at 7500 and 15,000 ppm, and the relative heart and adrenal weights were elevated at 15,000 ppm. There were no treatment-related clinical signs, gross autopsy or histopathological findings for either sex at any dose level. Animals improved only slightly from the effects of treatment following a 4-wk recovery period. A no-observable-effect level of 1000 ppm DETA.2HCl in the diet was established in this subchronic toxicity study. PMID- 9216748 TI - Differential prenatal toxicity of one straight-chain and five branched-chain primary alcohols in rats. AB - One straight-chain alcohol (n-octanol) and five branched-chain alcohols [2 ethylhexanol (2-EH), isodecanol, two types of isononanol and a C7-9-11 alcohol] were investigated for developmental effects in Wistar rats at equimolar dose levels (0, 1, 5 and 10 mmol/kg by gavage from gestation day 6 to 15; 10 animals per group). n-Octanol and both isononanols were also investigated in a supplementary experiment at 7.5 mmol/kg/day. Pronounced maternal but no developmental toxicity was achieved with n-octanol. C7-9-11 alcohol, which is a mixture of isomers mostly of a low degree of branching (alpha-methyl), showed no adverse effects at any dose levels. The two types of isononanols (typical mixtures of two different sets of isomers originating from two different production routes) exhibited a marked degree of maternal and foetal toxicity at 7.5 and 10 mmol/kg and slight foetal effects at 5 mmol/kg. Because of maternal toxicity in the top dose, a statistically significant increase in malformations was obtained only in the dose window of 7.5 mmol/kg in the supplementary experiment. Isodecanol (a mixture of different isomers) elicited maternal toxicity at 10 mmol/kg and caused a low incidence of retardations and rare malformations at that dose level. Some maternal signs but no foetal effects were observed at 5 mmol/kg. 2-EH showed strong maternal and also foetal toxicity at 10 mmol/kg and slight maternal and foetal toxicity at 5 mmol/kg. The differential responses to the test materials indicate that, at present, within this chemical class of alcohols, the potential for developmental toxicity has to be investigated case by case for each individual structure. PMID- 9216749 TI - Differential prenatal toxicity of branched phthalate esters in rats. AB - Several phthalate esters with alcohol moieties ranging from C5-C10 chains were investigated for prenatal toxicity in 10 rats per group after gavage administration of 0, 40, 200 and 1000 mg/kg/day from gestation day 6 to 15. At 1000 mg/kg di(2-ethylhexyl) phthalate showed clear foetotoxicity, embryolethality and teratogenicity. No significant effects were recorded at 40 and 200 mg/kg. Di 711-phthalate, a phthalic ester with linear components and a predominantly alpha methyl branching type, and di-isopentylphthalate showed a very similar spectrum of effects. Interestingly, the alcohol moiety of di-711-phthalate, 711-alcohol was developmentally inactive in a previous experiment. Di-iso-decylphthalate and three types of di-iso-nonylphthalate showed foetal effects of borderline significance at 1000 mg/kg/day. PMID- 9216750 TI - Human skin in vitro percutaneous absorption of gaseous ethylene oxide from fabric. AB - Ethylene oxide, a colourless gas at ordinary room temperature and pressure, is widely used as a fumigant, coming in contact with clothing and human skin. It is genotoxic in somatic and germ cells. [1,2-14C]Ethylene oxide and fabric discs were sealed in a glass container; the fabric discs were then removed and placed on human skin mounted in glass diffusion cells. Percutaneous absorption was 1.3% of the dose when the fabric/skin surface was open to surrounding air, and increased to 46.0% when the surface was occluded with latex glove material. The absorption was rapid, occurring within the first 0-4 hr assay period. Absorbed chemical was confirmed to be unchanged ethylene oxide in the receptor fluid. This study also serves as a model for exposure of fabric/skin to any potentially hazardous gas. PMID- 9216751 TI - Inhibition by wheat bran cereals of the development of aberrant crypt foci and colon tumours. AB - As variation in both type of fibre and its physical properties can influence physiological effects, the effects of different dietary levels (1, 4, 8%, w/w) of unprocessed wheat bran (WB) were compared with those of two of its processed commercial formulations used in breakfast cereals, on the formation of aberrant crypt foci (ACF) and colon tumours in Fischer 344 rats following azoxymethane (AOM) administration. All diets were high in fat (20 g/100 g) and low in calcium (0.2%, w/w). The rats were fed the experimental diets for 2 wk before receiving two sc injections of AOM (15 mg/kg body weight/wk). 8 wk following the first injection of AOM, five rats per group were killed and the formation of ACF was measured. 23 wk following the first injection of AOM, 12 rats per group were killed and the colon tumour incidence in different dietary groups was measured. The results showed that increasing the dietary concentration of fibre from 1 to 8% (w/w), using all the wheat bran formulations, significantly reduced the number of ACF per rat. None of the diets showed any significant effect on the normal growth of rats. No statistically significant differences were observed between the protective properties of WB and the two commercial formulations under investigation in terms of the reduction of the number of ACF, or in terms of the reduction of the colon adenocarcinoma incidence. The results suggest that wheat bran and its two commercial formulations can offer protection against colon cancer even when they are consumed with a high-fat/low-calcium diet. The addition of any of these formulations of wheat bran fibre is likely to be equally effective in the prevention of colon cancer in human populations that habitually consume high-fat/low-fibre Western-style diets. PMID- 9216752 TI - Assessment of mutagenic potential of thiram. AB - Thiram is a widely used dithiocarbamate fungicide. In this study, the mutagenicity of thiram was investigated using the micronucleus and dominant lethal tests in Swiss albino mice. A single ip injection of 100 mg thiram/kg body weight, which is the maximum tolerated dose (MTD), significantly induced micronucleus formation in bone marrow cells after 30 and 48 hr of exposure; 50% and 25% of the MTD also induced micronucleus formation after the above time periods. A significant number of dead implants were induced when thiram was given to male mice in the diet at 10% of the oral LD50 during the whole spermatogenesis cycle (8 wk); this post-implantation loss indicates a dominant lethal mutation. PMID- 9216753 TI - Suspected scaphoid fractures in skeletally immature patients: application of MRI. AB - PURPOSE: The purpose of our study was to evaluate the MR findings in the wrists of pediatric patients who have sustained acute wrist injuries and to determine if this imaging method yields more information than combined serial radiographs and physical examinations. METHOD: Eighteen skeletally immature patients (11 boys and 7 girls, age range 8-15 years) who had presented to the emergency room within 2 days following acute wrist trauma underwent serial clinical, radiographic, and MR examinations if there was a suspicion of a scaphoid fracture. RESULTS: Ten patients had a scaphoid abnormality on MR images. Six had fractures and four had regional bone marrow edema. Initially, all but two fractures were radiographically occult, although the other fractures eventually became evident on later studies. Those with marrow edema did not progress to fractures. Obliteration of the scaphoid fat stripe occurred in five patients with a scaphoid fracture and in six patients who did not have a fracture. Dorsal soft tissue swelling occurred in eight patients, five of whom had scaphoid fractures. Seven patients had evidence of extensor tenosynovitis on MRI. CONCLUSION: A normal initial MR image had a negative predictive value of 100%. Persistent snuffbox pain may represent injury to the scaphoid, extensor tendons, or dorsal soft tissues. An outcome study evaluating the benefits of early application of MR in the pediatric population is warranted. PMID- 9216754 TI - MR arthrography of the elbow: normal anatomy and diagnostic pitfalls. AB - PURPOSE: Our goal was to determine the normal MR arthrographic appearance of the elbow and to identify the potential diagnostic pitfalls of this technique. METHOD: MRI with intra-articularly administered gadopentetate dimeglumine was performed in seven elbow joints derived from cadavers. Coronal, sagittal, and axial SE T1-weighted images with fat suppression and coronal 3D SPGR images with fat suppression were obtained in each elbow. The elbows were then sectioned. RESULTS: This technique allowed depiction of the cartilaginous surfaces of the elbow joint as well as clear delineation of the inner surface of the collateral ligaments. Potential diagnostic pitfalls included fat pads projecting into the joint, synovial folds, and cartilaginous pseudoerosions. CONCLUSION: MR arthrography allows clear delineation of the intraarticular structures of the elbow. However, diagnostic pitfalls exist that increase the difficulty of interpreting MR arthrographic images. PMID- 9216755 TI - Popliteal artery entrapment syndrome: aberrant origin of gastrocnemius muscle shown by 3D CT. AB - PURPOSE: The purpose of this report is to evaluate the usefulness of 3D CT in the diagnosis of popliteal artery entrapment (PAE) syndrome. METHOD: Three patients (three men, 22-70 years old) with suspected PAE syndrome were examined using helical CT. 3D images were reconstructed to evaluate the relationship between the popliteal artery and the gastrocnemius muscle. 3D arteriograms were simultaneously reconstructed. RESULTS: One patient had unilateral type 3 PAE syndrome. Bilateral PAE syndrome was seen in the other two cases: One had bilateral type 2 anomalies and the other had type 3 on the left and type 2 on the right. In all three cases the anomalous origin of the medial head of the gastrocnemius muscle and its relationship to popliteal artery were clearly visualized by 3D CT. In addition, CT arteriography could detect occlusion, deviation, or stenosis of the popliteal arteries. CONCLUSION: 3D CT is useful in the diagnosis and management of PAE syndrome. PMID- 9216756 TI - Red marrow recolonization induced by growth factors mimicking an increase in tumor volume during preoperative chemotherapy: MR study. AB - Growth factors associated with chemotherapy may induce red marrow reconversion. This may simulate a tumor increase on MRI. We report such a case. PMID- 9216757 TI - Peripheral nerve stimulation by time-varying magnetic fields. AB - PURPOSE: A study was performed to assess the stimulation threshold for healthy adults using sinusoidally oscillating gradients. METHOD: One hundred thirteen healthy adults were examined in the study. ECG and physiological parameters were measured. All measurements were performed of both the head and the abdomen. The subjects were measured in the supine position with the region of interest positioned in the center of the gradient coils. The measurement was performed for three orthogonal, four oblique, and double oblique orientations. RESULTS: No volunteer reported painful, severe stimulation. The mean thresholds for peripheral stimulation in head and body measurement were similar: 85.5% of stimulation during examination of the head and 87.6% during measurements of the abdomen were reported when the y-gradient was used. CONCLUSION: The greatest frequency of reported stimulations occurs when the y-gradient is used. This was confirmed by the results and supports the hypothesis that orthogonal to the y axis the body has its largest conductive loop, resulting in the strongest peripheral stimulation. PMID- 9216758 TI - X-ray microtomography (microCT) using phase contrast for the investigation of organic matter. AB - PURPOSE: We show that microtomography (microCT) using synchrotron radiation (SR) can be extended to include X-ray phase contrast, which is two to three orders of magnitude more sensitive than conventional attenuation contrast and better suited for the investigation of specimens consisting chiefly of light elements for photon energies ranging at least from 1 to 100 keV. METHOD: Phase contrast is generated by placing the specimen in one of the interfering beams of an X-ray interferometer. With use of 12-keV X-rays, phase projections of the specimen are recorded at 180 or 360 angular settings equally spaced between 0 and 180 degrees. One phase projection consists of four pairs of "associated" radiograms in the sense that one is taken with and the other without the specimen in the beam. Between pairs a parallel-sided phase-shifter plate is rotated for changing the relative phase of the two interfering beams by multiples of pi/2 rad. By calculating phase-weighted sums of all associated pairs of radiograms, true phase shift projections are obtained for all angular settings of the specimen, which are then reconstructed. RESULTS: Three-dimensional images have been obtained from rat cerebrum and rat trigeminal nerve, showing cell structures at 8- to 15-micron spatial resolution. Gray and white matter of cerebrum and neurons in the trigeminal nerve are clearly visible. CONCLUSION: X-ray phase-contrast microCT is becoming a valuable tool for studies of organic samples in medicine and biology. PMID- 9216759 TI - Comparison and evaluation of retrospective intermodality brain image registration techniques. AB - PURPOSE: The primary objective of this study is to perform a blinded evaluation of a group of retrospective image registration techniques using as a gold standard a prospective, marker-based registration method. To ensure blindedness, all retrospective registrations were performed by participants who had no knowledge of the gold standard results until after their results had been submitted. A secondary goal of the project is to evaluate the importance of correcting geometrical distortion in MR images by comparing the retrospective registration error in the rectified images, i.e., those that have had the distortion correction applied, with that of the same images before rectification. METHOD: Image volumes of three modalities (CT, MR, and PET) were obtained from patients undergoing neurosurgery at Vanderbilt University Medical Center on whom bone-implanted fiducial markers were mounted. These volumes had all traces of the markers removed and were provided via the Internet to project collaborators outside Vanderbilt, who then performed retrospective registrations on the volumes, calculating transformations from CT to MR and/ or from PET to MR. These investigators communicated their transformations again via the Internet to Vanderbilt, where the accuracy of each registration was evaluated. In this evaluation, the accuracy is measured at multiple volumes of interest (VOIs), i.e., areas in the brain that would commonly be areas of neurological interest. A VOI is defined in the MR image and its centroid c is determined. Then, the prospective registration is used to obtain the corresponding point c' in CT or PET. To this point, the retrospective registration is then applied, producing c" in MR. Statistics are gathered on the target registration error (TRE), which is the distance between the original point c and its corresponding point c". RESULTS: This article presents statistics on the TRE calculated for each registration technique in this study and provides a brief description of each technique and an estimate of both preparation and execution time needed to perform the registration. CONCLUSION: Our results indicate that retrospective techniques have the potential to produce satisfactory results much of the time, but that visual inspection is necessary to guard against large errors. PMID- 9216761 TI - Intracranial developmental venous anomalies: diagnosis using CT angiography. AB - Four cases in which the diagnosis of developmental venous anomaly (DVA) was made using CT angiography are illustrated. The diagnosis of DVA was confirmed by either MR (all cases), digital subtraction angiography (two cases), or both (one case). This is the first report, to our knowledge, of the CT angiographic appearance of DVA. PMID- 9216760 TI - Detection and mapping of abnormal brain structure with a probabilistic atlas of cortical surfaces. AB - PURPOSE: We have devised, implemented, and tested a technique for creating a comprehensive probabilistic atlas of the human cerebral cortex, based on high dimensional fluid transformations. The goal of the atlas is to detect and quantify subtle and distributed patterns of deviation from normal cortical anatomy, in a 3D brain image from any given subject. METHOD: Given a 3D MR image of a new subject, a high-resolution surface representation of the cerebral cortex is automatically extracted. The algorithm then calculates a set of high dimensional volumetric maps, fluidly deforming this surface into structural correspondence with other cortical surfaces, selected one by one from an anatomic image database. The family of volumetric warps so constructed encodes statistical properties of local anatomical variation across the cortical surface. Additional strategies are developed to fluidly deform the sulcal patterns of different subjects into structural correspondence. A probability space of random transformations, based on the theory of anisotropic Gaussian random fields, is then used to encode information on complex variations in gyral and sulcal topography from one individual to another. A complete system of 256(2) probability density functions is computed to reflect the observed variability in stereotaxic space of the points whose correspondences are found by the warping algorithm. Confidence limits in stereotaxic space are determined for cortical surface points in the new subject's brain. RESULTS: Color-coded probability maps are generated, which highlight and quantify regional patterns of deformity in the anatomy of new subjects. These maps indicate locally the probability of each anatomic point being as unusually situated, given the distributions of corresponding points in the scans of normal subjects. 3D MRI volumes are analyzed, from subjects with clinically determined Alzheimer disease and age matched normal subjects. CONCLUSION: Applications of the random fluid-based probabilistic atlas include the transfer of multisubject 3D functional, vascular, and histologic maps onto a single anatomic template, the mapping of 3D atlases onto the scans of new subjects, and the rapid detection, quantification, and mapping of local shape changes in 3D medical images in disease and during normal or abnormal growth and development. PMID- 9216762 TI - Intracranial aneurysm: inner view and neck identification with CT angiography virtual endoscopy. AB - We describe a virtual endoscopy tool applied on CT angiography acquisitions to analyze the neck and inner structures of an intracranial aneurysm. This technique, applied on a basilar artery aneurysm, accurately described its morphology and helped in making a choice between surgical and endovascular treatment. PMID- 9216764 TI - CT demonstration of an unusual aspect of the thyroid gland. PMID- 9216763 TI - PET for diagnosis of malignant lymphoma of the scalp: comparison of [11C]methyl-L methionine and [18F]fluoro-2-deoxyglucose. AB - A 43-year-old woman was admitted with a tumor mass in her forehead. Two months previously, a lump in her breast had been diagnosed as mastopathy. Palpation revealed an elastically hard immobile tumor mass in her forehead. MRI detected a tumoral lesion of generally uniform contrast involving frontal subcutaneous, cranial, and intracranial regions. PET demonstrated more intensive and wider accumulation of [11C]methyl-L-methionine (Met) than of [18F]fluoro-2-deoxyglucose (FDG). Biopsy of the forehead mass was performed, which was diagnosed as B-cell type malignant lymphoma. The tumor mass in the forehead then shrank spontaneously, as confirmed by palpation and MRI. The tumor mass in the left breast was totally extirpated and histologically diagnosed as B-cell-type malignant lymphoma, like the tumor mass in the forehead. Postoperatively, chemotherapy (VEPA) was performed. Although FDG accumulation had not been detected, postchemotherapy PET demonstrated slight Met accumulation, suggesting the presence of a residual tumor. PET served well to detect the lesion and evaluate therapeutic efficacy in malignant lymphoma. Met-PET was more sensitive to malignant lymphoma than FDG-PET. PMID- 9216765 TI - Bronchogenic carcinoma invasion of the chest wall: evaluation with dynamic cine MRI during breathing. AB - PURPOSE: Our goal was to assess the utility of breathing dynamic cine MR (BDCMR) in the evaluation of tumor invasion to the chest wall in bronchogenic carcinomas. METHOD: BDCMR imaging was performed preoperatively in 25 patients with bronchogenic carcinomas adjacent to the chest wall. Twelve sequential images were obtained in the same coronal and/or sagittal planes during one respiratory cycle with fast spoiled GRASS sequence, and analysis with cine-loop display was performed. RESULTS: In all 14 cases in which free movement of tumor along the parietal pleura on BDCMR was demonstrated, no chest wall invasion was evident at thoracotomy. However, of 11 patients with fixation of the tumor on BDCMR, 5 had benign pleural adhesions, 5 had chest wall invasion at thoracotomy, and 1 with an apical tumor had no benign pleural adhesion or chest wall invasion. CONCLUSION: Although BDCMR cannot distinguish benign pleural adhesions from chest wall invasion by tumor, this method accurately estimated the free movement of lung tumors with no invasion of chest wall from bronchogenic carcinomas prior to surgery. PMID- 9216766 TI - High resolution chest CT in tuberculosis: evolutive patterns and signs of activity. AB - PURPOSE: The purpose of our study was to determine evolutive patterns and signs of active tuberculosis on high resolution CT (HRCT) scans. METHOD: We followed up over 15 months 27 patients with postprimary pulmonary tuberculosis that was proven bacteriologically. CT scans were performed before, during, and after 6 months of anti-tuberculosis treatment. Both 10-mm-thick sections and 1.5-mm-thick HRCT scans were performed. RESULTS: Ground-glass pattern was noticed 26 times, 9 times after 2 month treatment and only 2 times after 6 month treatment. Among these two patients, one did not undergo his treatment properly and the other one had an additional bacterial infection. Centrilobular nodules (n = 17) and poorly marginated nodules (n = 21) were present only before treatment. Reticular pattern (intralobular and septal thickening), interstitial nodules, and fibrosis were seen both before and after treatment. Ground-glass pattern, poorly marginated nodules, and infiltrates as well as centrilobular nodules were related to an active infection. CONCLUSION: This HRCT may be helpful to demonstrate activity in patients suspected of having tuberculosis and to assess antituberculous treatment efficiency. PMID- 9216767 TI - Intrathoracic lymphadenopathy in patients with empyema. AB - PURPOSE: Our goal was to determine the prevalence of intrathoracic lymphadenopathy on chest CT in patients with empyema. METHOD: We retrospectively identified 27 patients (14 men, 13 women, mean age 43 years) with nontuberculous empyema examined with chest CT. All scans were reviewed by two of three board certified radiologists for the presence of intrathoracic lymphadenopathy (> or = 1 cm, short axis) in an American Thoracic Society (ATS) nodal station or the internal mammary region. Differences were resolved by consensus. RESULTS: Thirteen (48%) patients with empyema had lymphadenopathy on chest CT. The mean number of enlarged lymph nodes for the patients with lymphadenopathy was 3.2 (SD +/-2.3, range 1-8). The mean size of the largest lymph node was 1.4 cm (range 1.0 2.5 cm). The lymphadenopathy was unilateral and ipsilateral to the empyema in seven (54%), bilateral in five (38%), and unilateral contralateral to the empyema in one. The distribution of lymphadenopathy according to ATS nodal stations was 4R (n = 8), 7 (n = 6), 10R (n = 5); n = 2 each 2R, 10L, 11L; and n = 1 each 11R, 2L, 4L, and 6. Four patients had internal mammary lymphadenopathy. Pleural fluid and smooth pleural thickening were present in each case. Four patients had follow up CT after treatment. There was a decrease or resolution of the lymphadenopathy in each case. CONCLUSION: Intrathoracic lymphadenopathy is a common CT finding in patients with empyema and occurred in 48% of this series. In patients with smooth pleural thickening and pleural effusion, intrathoracic lymphadenopathy should not be used as a criterion to differentiate empyema from malignant or tuberculous pleural effusion. PMID- 9216768 TI - Pulmonary involvement of idiopathic hypereosinophilic syndrome: CT findings in five patients. AB - PURPOSE: The aim of this study was to assess the CT findings of pulmonary involvement in patients with idiopathic hypereosinophilic syndrome (HES). METHOD: The study included five patients with idiopathic HES who had pulmonary involvement proven by bronchoalveolar lavage (n = 3) or based on clinical and radiologic findings (n = 2). Four patients had high resolution CT and one had conventional CT. The CT scans were retrospectively reviewed by two chest radiologists for pattern and distribution of disease. RESULTS: All five patients had several small nodules in both lungs at CT scan. Four patients had nodules with a halo of ground-glass attenuation. Three patients had focal areas of ground glass attenuation in both lungs. These lesions were present in all lung zones and involved mainly the peripheral lung. There was neither lobar predilection nor peribronchovascular distribution. Other organs involved included bone marrow (n = 3), liver (n = 3), stomach (n = 1), and peritoneum (n = 1). CONCLUSION: The CT findings of pulmonary involvement in patients with idiopathic HES included small nodules with or without a halo of ground-glass attenuation and focal areas of ground-glass attenuation mainly in the lung periphery. PMID- 9216769 TI - Swyer-James syndrome documented by spiral CT angiography and high resolution inspiratory and expiratory CT: an accurate single modality exploration. AB - Spiral CT angiography was performed in a patient suspected of having pulmonary embolism. The right pulmonary system was normal. The left arterial system was small but patent. The left upper lobe was small and hyperlucent. The left lower lobe was collapsed and contained bronchiectasis. The bronchi were patent. High resolution CT in inspiration and expiration confirmed air trapping in the left upper lobe. A diagnosis of Swyer-James syndrome of the left upper lobe was made. PMID- 9216770 TI - Coronary artery calcification at CT as a predictor for cardiac complications of thoracic surgery. AB - PURPOSE: Our goal was to determine the predictive value of coronary artery calcification (CAC) on preoperative CT of the thorax for cardiac complications of noncardiac thoracic surgery. METHOD: Of 117 patients undergoing noncardiac thoracic surgical procedures between January 1, 1993, and June 1, 1995, at our institution, 75 had inpatient records and chest CTs available for retrospective review. Inpatient records were reviewed for postoperative cardiac complications (arrhythmia, hypotension with ECG changes, myocardial infarction, congestive heart failure, stroke, and death). The CT scans were scored for the presence and extent of CAC by an independent observer. RESULTS: Six of the 75 patients had cardiac complications including 1 death. Thirty-nine of the 75 patients had a CAC score of > or = 7. The sensitivity, specificity, positive predictive value, and negative predictive value of a CAC score of > or = 7 for cardiac complications were 100, 71, 23, and 100%, respectively. CONCLUSION: The presence of CAC on preoperative CT scanning is associated with cardiac complications of noncardiac thoracic surgery; however, the positive predictive value is low. The absence of CAC was a reliable predictor of a favorable postoperative cardiac course. PMID- 9216771 TI - Venous aneurysms: MR diagnosis with the "layered gadolinium" sign. AB - OBJECTIVE: Our goal was to present MR findings in venous aneurysms and introduce the "layered gadolinium" sign as an ancillary diagnostic finding. METHOD: Gadolinium-enhanced MR images of three patients with retroperitoneal venous aneurysms were retrospectively reviewed. Prior to MRI, venous aneurysm had been suspected clinically in only one patient. Surgical correlation was available in one patient. A phantom was constructed and imaged to investigate the cause of the layered gadolinium sign. RESULTS: A gradation of signal intensity, the layered gadolinium sign, was observed in three patients with venous aneurysms on postcontrast T1-weighted images. The anterior portion of the aneurysms demonstrated high signal intensity separated by a sharp interface from the low signal intensity posterior region. Unenhanced time-of-flight MR venography, color Doppler, and duplex sonography failed to demonstrate flow in the patient with surgical proof. CONCLUSION: The layered gadolinium sign may be helpful in the diagnosis of venous aneurysm and in differentiating these masses from solid neoplasms. PMID- 9216772 TI - Scimitar syndrome: MR assessment of hemodynamic significance. AB - We report a case of anomalous pulmonary venous drainage into the inferior vena cava (scimitar syndrome) in which the hemodynamic significance was noninvasively assessed by means of velocity-encoded cine MR. Left-to-right shunt was calculated from direct blood flow measurements performed in the ascending aorta, main pulmonary artery, and aberrant pulmonary vein. PMID- 9216774 TI - Spiral CT in the evaluation of flank pain: overall accuracy and feature analysis. AB - PURPOSE: Our goal was to assess test reliability and identify those features that have the strongest positive and negative predictive values in the diagnosis of renal colic using spiral CT. METHOD: Fifty non-contrast-enhanced CT scans (5 mm slice thickness) obtained in patients presenting with flank pain were reviewed by three radiologists blinded to the final diagnoses. The sensitivity, specificity, and positive and negative predictive values for nine pertinent findings were determined as compared to clinical follow-up. RESULTS: Twenty-nine cases had findings of ureteral obstruction. Findings with the strongest positive predictive values (> 0.90) were ureteral stone, hydronephrosis, hydroureter, periureteral stranding, and ureterovesical junction edema. Findings with the strongest negative predictive values (> 0.89) were absence of hydronephrosis and hydroureter. The areas under the receiver operating curves for Readers 1, 2, and 3 were 0.970 +/- 0.030, 0.942 +/- 0.036, and 0.982 +/- 0.020. CONCLUSION: Absence of hydroureter and hydronephrosis on spiral CT images should prompt a search for a diagnosis other than an obstructing ureteral stone. PMID- 9216773 TI - Dynamic breath-hold 3D gadolinium-enhanced MRI of intraarterial masses: findings in two patients. AB - We describe the use of dynamic breath-hold gadolinium-enhanced 3D MRI for the evaluation of intraarterial masses and its potential use in differentiating thrombus from neoplasm. This rapid technique overcomes the flow artifacts that may obscure intraarterial masses on contrast-enhanced ECG-gated SE MR and can be performed on patients who otherwise cannot be ECG gated. PMID- 9216775 TI - Tuberculosis of the prostate: MR appearance. AB - Tuberculosis of the prostate is uncommon. Here we present a case of tuberculosis of the prostate with its MR findings. PMID- 9216776 TI - Proximal ureter obstruction caused by a lower polar renal artery: demonstration with spiral CT angiography. PMID- 9216777 TI - Dynamic MRI using a surface coil in chronic cholecystitis and gallbladder carcinoma: radiologic and histopathologic correlation. AB - PURPOSE: The purpose of this study was to determine whether dynamic MRI could differentiate gallbladder carcinoma from chronic cholecystitis. METHOD: The dynamic MR findings of 50 patients with pathologically proven chronic cholecystitis and 13 with gallbladder carcinomas were correlated with the pathological findings. RESULTS: In chronic cholecystitis with thickened wall, mucosa and muscle were shown in early images as smoothly delineated enhancement except in one case, and the subserosa with fibrosis was enhanced in late or delayed images. Unenhanced foci in the wall correlated with Rokitansky-Aschoff sinuses or mural stones. In carcinomas, all tumors showed irregularly delineated enhancement in early images. In late or delayed images, noncancerous portions were also enhanced. The outer margin of early enhancement correlated with the extension of the tumor. CONCLUSION: Dynamic MRI is useful for the differentiation of chronic cholecystitis from carcinoma and for the evaluation of its local extension. PMID- 9216778 TI - CT in searching for abscess after abdominal or pelvic surgery in patients with neoplasia: do abdomen and pelvis both need to be scanned? AB - PURPOSE: This prospective study was undertaken to determine the incremental yield of combined abdominal and pelvic CT in searching for clinically suspected postoperative abscess in oncologic patients. METHOD: One hundred seventeen oncologic patients underwent CT to exclude a clinically suspected abscess within 30 days of abdominal or pelvic surgery during an 8 month period. Scans were evaluated for the presence of ascites, loculated fluid collections, or other possible sources of fever. The clinical course and any intervention in the abdomen or pelvis within 30 days after CT were recorded. RESULTS: After abdominal surgery, 44 of 69 [64%; confidence interval (CI) 51-75%] patients had loculated fluid collections in the abdomen; no patient (0%; CI 0-5%) had a loculated fluid collection present only in the pelvis. After pelvic surgery, 22 of 48 (46%; CI 31 61%) patients had loculated fluid collections in the pelvis; no patient (0%; CI 0 7%) had a loculated collection present only in the abdomen. Loculated collections were present in both the abdomen and the pelvis in 4 of 69 (6%; CI 1.6-14%) patients after abdominal surgery and 3 of 48 (6%; CI 1.3-17%) after pelvic surgery. CONCLUSION: Isolated pelvic abscesses after abdominal surgery and isolated abdominal abscesses after pelvic surgery appear to be very uncommon in oncologic patients. CT initially need be directed only to the region of surgery in this particular patient population. PMID- 9216780 TI - CT of appendicitis in children. AB - PURPOSE: Our goal was to review the CT findings and to help define the role of CT in the evaluation of appendicitis in children. METHOD: Of 730 children with surgically proven appendicitis, 22 underwent preoperative CT evaluation. Their CT scans and operative and pathology records were retrospectively reviewed. The CT scans were evaluated for appendiceal wall thickness, diameter, and location, appendicoliths, pericecal inflammation, phlegmon, abscess, free fluid, small bowel dilatation, and bowel wall thickening. Criteria for diagnosing appendicitis were (a) appendiceal wall thickening (> 1 mm) or (b) presence of abscess, phlegmon, or pericecal inflammation associated with appendicolith(s). Prospective reports of ultrasound examinations performed within 2 days of the CT scans were available in 14 children and were correlated with the CT findings. RESULTS: An abnormally thickened appendix, with a diameter ranging from 9 to 18 mm, was seen in four children. Three appendices were retrocecal and one was near the cecal tip, anterior to the iliac vessels. Appendicoliths were present in 10 children, multiple in 1. Abscesses were seen in 13 of 22 children, multiple in 5. Phlegmon was seen in five children and pericecal inflammation in two. Bowel wall thickening was present in seven children and small bowel dilatation was noted in six. Other findings included free fluid, hydronephrosis, thickening of urinary bladder wall, air in the uterus and vagina, adenopathy, and thickening of the abdominal wall musculature. CT was diagnostic of appendicitis in 11 of 22 children (50%). In 14 children with both ultrasound and CT studies, CT was slightly better in diagnosing appendicitis and visualizing the abnormal appendix and was superior in defining the presence and extent of abscess and inflammation in 9 of 14 children. CONCLUSION: CT is a useful adjunct in diagnosing appendicitis in children, with a major role in cases of complicated appendicitis. PMID- 9216779 TI - Liver metastases from colon cancer with intra-bile duct tumor growth: radiologic features. AB - PURPOSE: Our goal was to characterize the radiologic features of liver metastases from colon cancer with intrahepatic bile duct (IHBD) dilatation. METHOD: Radiologic findings of liver metastases from colon cancer with IHBD dilatation of four patients were compared with pathologic findings. RESULTS: The cause of bile duct dilatation in all cases was due to papillary tumor growth in the bile duct. In two patients, intra-bile duct tumor growth (IBDTG) was observed on imaging. In the other two patients, IBDTG was not observed, but a nontapered abrupt obstruction of a dilated bile duct was seen, corresponding to the microscopically proven papillary tumor growth in the ductal lumen. In three patients who underwent an extensive hepatic resection, there has been no recurrence. In one patient who had a nonanatomic limited resection, a recurrence was seen 1 year after surgery. CONCLUSION: When liver tumor with IBDTG is suspected on imaging, liver metastases should be considered in the differential diagnosis besides hepatocellular carcinoma or cholangiocellular carcinoma. Careful preoperative assessment for IBDTG by imaging is essential to determine the extent of surgical resection. PMID- 9216781 TI - Continuation of gas from the right perirenal space into the bare area of the liver. AB - PURPOSE: It is difficult to explain some CT findings pertaining the anatomic relationship between the bare area of the liver and the right retroperitoneal space on the basis of the traditional three compartment concept of retroperitoneal anatomy. The purpose of this article is to ascertain and illustrate possible communication between the bare area of the liver and the right perirenal space in patients with gas-forming retroperitoneal infection. METHOD: We reviewed CT findings in 24 cases with retroperitoneal gas-forming condition with particular emphasis on the extent of the infection and resultant distribution of gas. RESULTS: CT images showed gas within the right perirenal space extended upward directly into the bare area of the liver in six cases. CONCLUSION: The right perirenal space is considered open toward the bare area of the liver. Therefore, gas within the perirenal space may communicate directly with the bare area of the liver. PMID- 9216782 TI - Ossification of a rectal tumor: CT evaluation. AB - Adenocarcinoma of the colon is a common malignancy. Not infrequently, this tumor can calcify. An extremely rare occurrence in these tumors is ossification. This is the first reported case of rectal carcinoma in which ossification was demonstrated with CT. PMID- 9216783 TI - Colosplenic fistula in a patient treated with interleukin-2 for malignant melanoma. AB - Interleukin-2 (IL-2) therapy often causes gastrointestinal side effects and at least 8 cases of bowel perforation have been reported. The patient reported here developed a colosplenic fistula, diagnosed by CT, with no neoplastic involvement of these organs. Awareness of these complications of IL-2 can help lead to earlier diagnosis. PMID- 9216784 TI - Macronodular hepatic granulomas due to visceral leishmaniasis in an AIDS patient: imaging findings. PMID- 9216785 TI - Aunt Minnie's corner. Pulmonary hamartoma. PMID- 9216786 TI - Antimutagenesis and anticancer effects. AB - Cancer development is a multistage process wherein mutational events play an important role. Various antimutagen components, in particular magnesium and selenium, have been reported to have anticarcinogenic properties. These two oligoelements display different behaviors according to their concentrations. The different effects of magnesium and selenium depend on the different mechanisms of cell absorption. The importance of antimutagenesis studies in cancer prevention is reported and a review of the basic mechanisms operative in antimutagenesis, including some data on known antimutagens, is made. PMID- 9216787 TI - Potassium, sodium, and cancer: a review. AB - Agents known or believed to be carcinogenic decrease the concentration of potassium and increase the concentration of sodium in the cells. Anticarcinogenic agents have the opposite effect. In all cases where we have information on an agent's carcinogenicity or anticarcinogenicity and on that agent's effects on cellular potassium and sodium concentrations the above relationships have been found to be true. Dietary carcinogenic agents studied include sodium, cadmium, fat, cholesterol, calories, and alcohol; dietary anticarcinogenic agents include potassium, vitamins A, C, and D, selenium, and fiber. The effect of calcium intake is less clear as that effect depends on the concentrations on sodium and potassium. Not only dietary agents but also other carcinogenic and anticarcinogenic agents work in the same way. The cancer-causing drug dimethylhydrazine increases sodium and decreases potassium in the cells, whereas, for example, indomethacin, an anticarcinogen, has the opposite effect. In aging potassium leaves the cells, sodium enters them, and the rates of cancer increase. Patients with hyperkalemic diseases (Parkinson, Addison) have reduced cancer rates, and patients with hypokalemic diseases (alcoholism, obesity, stress) have increased cancer rates. PMID- 9216788 TI - Genotoxic effects of pesticides. AB - Epidemiologic data showed an increase in the number of cancer cases in persons involved in agricultural production using pesticides. According to IARC, more than 25% of pesticides are classified as oncogens. In recent years, the concept of malignant tumors developing after environmental contamination with chemicals has been accepted. Changes in genetic material are at the basis of this process because many environmental pollutants are chemical carcinogens and mutagens with the capacity of causing DNA damage. DNA damage was proposed as a useful parameter for assessing the genotoxic properties of environmental pollutants. The correlation between exposure to carcinogenic substance and the level of DNA damage is essential. Pesticides are highly biologically active chemicals. They may interact with DNA and damage its structure. Such interaction may be critical for the manifestation of carcinogenic properties of different chemicals. We report on the organotropic genotoxic effects of different chemical classes of pesticides (decis, cypermetrin, 2,4-D, polyram) studied by means of alkaline unwinding assay DNA. PMID- 9216789 TI - The cytogenetic effects of pyrimethamine on male mouse germ cells. AB - Pyrimethamine is an inhibitor of dihydrofolate reductase and is used in the treatment of malaria and toxoplasmosis. We examined the cytogenetic effects of this drug. Adult male mice were given doses of 20, 40, 80, and 120 mg/kg pyrimethamine intraperitoneally. Animals were killed by cervical dislocation on the 3rd, 6th, 9th, and 12th day after treatment, and the primary spermatocytes were harvested from their testes. These cells were analyzed for gaps, breaks, acentric fragments, and exchanges, as well as for numerical aberrations such as univalency. A dose-related increase in chromosomal aberrations was found in the pyrimethamine group compared with the control group. We suspect that pyrimethamine is a possible clastogen that may affect human germ cells. PMID- 9216790 TI - Hepatitis C virus genotypes in patients with hepatocellular carcinoma. AB - OBJECTIVES: Epidemiological studies have suggested a strong association between chronic hepatitis C virus (HCV) infection and the development of hepatocellular carcinoma (HCC). It has also been suggested that there is a relationship between HCV genotypes and the development of cirrhosis and HCC. To investigate the possible role of HCV genotypes in the development of HCC, we studied HCV genotypes in 22 HCV-seropositive patients with histologically proven cirrhosis of the liver and HCC. METHODS: Anti-HCV antibodies were detected by second generation enzyme-linked immunosorbent assay (ELISA). Serum HCV-RNA was detected by nested reverse transcription-polymerase chain reaction (RT-PCR) using primers derived from the highly conserved 5'-untranslated region. The HCV genotype was determined by nested RT-PCR using type-specific primers derived from the core region. RESULTS: Anti-HCV and HCV-RNA were detected in all patients with HCC. HCV genotyping was achieved in all of them. All patients had genotype 1b HCV. CONCLUSION: These findings suggest that HCV remains in replication and genotype 1b HCV is the predominant type in our HCV-seropositive patients with HCC. PMID- 9216791 TI - Carotenoids in cancer, mastalgia, and AIDS: prevention and treatment--an overview. AB - In 1980, two carotenoids, beta-carotene (BC) and canthaxanthine (CX) with and without pro-vitamin A activity, respectively, were orally administered to female Swiss albino mice and were found to substantially prevent skin carcinogenesis induced by benzo(a)pyrene (BP). This preventive effect was observed in darkness by means of photocarcinogenic enhancement (PCE) following UV (300 to 400 nm) irradiation. In 1984, the same experiment produced antitumorigenic activity when applied to breast carcinogenesis induced in mice by 8-methoxypsoralen (8-MOP) plus UV-A light and, in 1985, when directed toward gastric carcinogenesis induced in rats by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). These data suggested a rationale for human intervention to prevent, by carotenoid supplementation, a second primary malignancy after the primary malignancy has been radically excised. In the 1980s, a pilot clinical study (15 cases) showed a longer than expected disease-free interval in all surviving patients. It was also subsequently found that, if treated daily with 20 mg of BC and intermittently with retinol 150 to 300,000 IU daily for seven days just prior to menses, women suffering from cyclical mastalgia were relieved from pain, without any toxic side effects. When BC was given in high daily doses (60 mg) to 60 drug addicts suffering from AIDS-related complex (ARC), they recovered from their objective and subjective symptoms (but not from lymphadenopathy) with improvement in their general health and increased performance status. At higher doses, BC (with or without hyperthermia) was effective even in patients in advanced stages of AIDS. A debate has arisen concerning a recent statement by the U.S. Government that "beta-carotene supplements do not protect Americans against cancer or heart disease, and may actually increase the risk of deadly lung tumors in smokers". PMID- 9216792 TI - Environment, cancer, and molecular epidemiology: air pollution. AB - Most cancers result from human interaction with the environment. As may be expected, air pollution is the most frequent factor responsible for environmental carcinogenesis due to natural exposures (such as air contamination, background radiation, and asbestos) or man-made pollution (e.g., smoking). A challenging problem in clinical epidemiology has been the nonuniform distribution of cancer among populations equally exposed to carcinogenic circumstances. Recent findings made available through the development of molecular biology techniques have provided new insights into cancer susceptibility. The wide variations in the uptake and ability to activate xenobiotics are key phenomena in environmental carcinogenesis. The intracellular DNA repair systems are probably responsible for the end result of neoplastic transformation or normalcy in the presence of carcinogenic encounters. PMID- 9216793 TI - Environmental pollution and carcinogenic risk. AB - In this study, the relationship between environmental pollution and environmental carcinogens has been investigated. There is an alarming increase of diverse natural and man-made carcinogens in consumer goods and industrial wastes. Research carried out in Ankara and Istanbul has found detergents and arsenic in the drinking water to be 140 times higher than the World Health Organization (WHO standards. In addition, air pollutants such as sulfur dioxide, particles, and fumes have been determined to be 4 times higher than the WHO standards. A similar situation has been noted in the soil pollution of the country. Pesticides and insecticides used in agriculture are seriously harming human health. It is necessary that the government and institutions establish effective controls and protective measures and educate the people. PMID- 9216794 TI - Atmospheric pollutants in Uludag National Park. AB - Gas phase pollutants and major ions were measured in the Saryalan region of Uludag Mountain, which is experiencing severe deforestation. Short- and long-term trends in the concentration of the gas pollutants were computed from the samples obtained in order to identify the source of these pollutants. The higher concentration of O3 during summer months was consistent with the higher photochemical production from precursor gases (NOx) with increased solar flux. The diurnal pattern of O3, NO, NO2 and TSP describes a photochemical smog scenario. A lower pH in aerosols associated with high levels of SO4(2-) and NO3- is an indication of acid deposition. Wind sector analysis revealed that the major contributing source regions are north and south of the sampling site, that is, the city of Bursa, local industries, and the Orhaneli power plant. PMID- 9216795 TI - Air pollution profile of Bursa. AB - Rapid urbanization and industrial development are the most important causes of air pollution in Bursa. Smoke and sulfur dioxide concentrations were measured at five stations over a period of 20 months between 1986 and 1987; the concentrations of the total suspended particles were determined in the samples collected at two stations in June and October 1986. Some of the trace elements (Fe, Pb, Cd, Zn) were measured in October 1988 by atomic absorption spectroscopy of 28 samples from two stations. The first-order regression equations were calculated in order to find the relationship between the concentrations of smoke, sulfur dioxide, and meteorological conditions. The trends in the concentrations of measured air pollutants were compared by the long- and short-term limit values, as specified in the regulation. PMID- 9216797 TI - Mapping in epidemiological studies and control of cancer: experiences from Slovakia. AB - This article reviews our experience with the use of the cancer atlas of Slovakia, published in 1989 which presented simultaneously the cancer incidence and mortality rates derived from national population-based cancer registries. Contrary to all expectations, the role of the environmental pollution was confirmed only for nonmelanoma skin cancer and arsenic exposure. Valuable information was obtained also for the study of dynamics of cancer distribution, which revealed a shift in the incidence of stomach cancer to the east and its replacement with colorectal cancer in the western part of the country. Of enormous importance for the comprehensive cancer care and control is the knowledge of incidence and mortality rates at the level of individual districts. PMID- 9216796 TI - Environmental and health monitoring in Lithuanian cities: exposure to heavy metals and benz(a)pyrene in Vilnius and Siauliai residents. AB - The Environmental and Health Monitoring program in the large cities of Lithuania is aimed at the evaluation of the population health status in terms of chemically induced diseases. During the 1991 to 1995 period, this program was carried out in two Lithuanian cities, Vilnius, the capital of the country, and Siauliai. Data on the chemical pollution of ambient air, soil, and drinking water and the morbidity were mapped. Risk zones of environmental pathology threat were defined within each city on the basis of the mapped data. Subsequently, chemical pollutants, namely, heavy metals and benz(a)pyrene, were determined in the biomedia of selected population groups in the risk zones. Exposure analysis of heavy metals (Pb, Cd, Ni, Cr, Cu, Zn) and benz(a)pyrene was carried out for standardized groups of children and pregnant women in the risk zones and in a relatively safe (control) zone. The evaluation of exposure to heavy metals was based on the levels found in blood, urine, and hair. Benz(a)pyrene was tested in urine samples. The obtained data are applied in the process of environmental health monitoring in the large cities of Lithuania. PMID- 9216798 TI - Variations in avoidable mortality in relation to health care resources and urbanization level. AB - "Avoidable" mortality may be defined as causes of death whose occurrence is closely related to medical intervention. Areas with particular health care delivery problems can be identified through a geographical comparison of these "avoidable deaths." Mortality data for Valencia from 1982 to 1990 were examined to determine whether or not the availability of medical care resources in the area influenced the occurrence of avoidable deaths. We identified variations in mortality from avoidable causes, grouped according to the differences in levels of urbanization and health care resources, in the 537 municipalities of the Valencian community. (In Spain, the municipality is the lowest administrative division.) Linear regression analysis was performed to predict or estimate this relationship. Only in a small number of avoidable causes did the mortality trend for males differ significantly from 0 (p < 0.005) in relation to different levels of urbanization and health care resources. A direct association between these two variables was observed in males with regards to pneumonia, tuberculosis, chronic rheumatic heart disease, and bacterial infection. In females, a relationship between "avoidable" mortality rates and the differences in urbanization and health care resources was found in cervical cancer, pneumonia, abdominal hernias, and cholecystitis. Mortality from asthma and cardiovascular disease (in both males and females) declined faster in urbanized, high income areas than in rural areas. The results clearly demonstrate the considerable mortality risk associated with living in urban areas. On the contrary, we found very little correlation between health service access and mortality. PMID- 9216799 TI - Tobacco and cancer in Turkey. AB - Although the smoking epidemic is decreasing steadily in other parts of the world, it continues to spread at an accelerated rate in underdeveloped and developing countries. Turkey, among other developing countries, faces the increasing threat of tobacco-related cancers, particularly lung cancer, which is the leading cause of cancer death in both sexes. We investigated the relationship between cigarette consumption and the relative mortality rates due to lung cancer in men and women between 1965 and 1992. We found a parallelism between the increasing total and per capita cigarette consumption and the rising relative mortality from lung cancer in both sexes. Total per capita cigarette consumption rose from 1230 cigarettes per year in 1985 to 1495 in 1991, and the per capita yearly cigarette consumption over the age of 15 increased from 1850 in 1965 to 2600 in 1992. During the same period, the relative mortality from lung cancer increased from 25 to 40% in men and from 11 to 16% in women. The tar, nicotine, and carbon monoxide determinations of locally produced and imported cigarettes suggested that the high tar and carbon monoxide content of most locally produced cigarettes smoked over many years could also be a contributory factor to the increased mortality rates due to lung cancer. Only two brands of locally produced cigarettes contained lower than 12 mg of tar per cigarette as allowed in European community states, whereas half of the imported brands of cigarettes met this standard. Four of the six imported brands of cigarettes contained higher tar and carbon monoxide compared with the same brands sold in England. These findings indicate that urgent measures are necessary not only to ban all activities promoting the sale of cigarettes but also to establish standards for both national and foreign brands of cigarettes while making a greater effort to reduce active and passive smoking in the Turkish population. PMID- 9216800 TI - Cigarette smoking, serum lipids, folate, and vitamin B12 in lung cancer. AB - The purpose of our study was to evaluate the effects of cigarette smoking and serum lipids, folate, and vitamin B12 on the development of lung cancer in the Turkish population. The study group consisted of patients with histologically proven lung cancer and the control group comprised healthy smokers being followed in our smoking cessation outpatient department. Smoking history was obtained from all subjects and serum total cholesterol, HDL cholesterol, triglycerides, vitamin B12, and folate levels were measured. Pack/years of cigarettes smoked were significantly higher in the subjects with lung cancer than in the control group (p < 0.01). Serum total cholesterol, HDL cholesterol, triglyceride, serum folate, and vitamin B12 levels were within normal limits in both groups (p < 0.05), but serum vitamin B12 levels were statistically significantly higher (p < 0.01) in the cancer group than in the controls. In our study, we did not observe low levels of serum cholesterol, vitamin B12, or folate in the lung cancer patients. PMID- 9216801 TI - The smoking addiction of pregnant women and the consequences on their offspring's intellectual development. AB - Many scientists have studied the effects of smoking by pregnant women on intrauterine development. Because nicotine and other toxic substances in cigarette smoke are not stopped by the placental barrier, there is a risk that the development of the child could be hindered. It has been shown, for instance, that babies whose mothers smoked during pregnancy have lower size and weight at birth. Few authors have studied the consequences a mother's pre-natal smoking may have on the intellectual development of her child. We compared two samples of children, aged 4 to 5, and aged 6 to 7 (40 children in total), whose mothers had smoked during pregnancy, with two samples of 40 children of the same ages whose mothers had not smoked. We tested them on the Wechsler scale. The social and cultural levels were equivalent. We found a difference of more than 15 IQ points in favor of the children of nonsmoking mothers. These results permit us to suppose that smoking during pregnancy hinders the intellectual development of the child. PMID- 9216802 TI - Subliminal manipulation of smoking. AB - Subliminal advertising techniques have increased in usage and are commonly accepted, particularly regarding cigarette smoking. Considering the high cost of such subliminal methods, their use can be justified only by tangible results, measured by an increase in cigarette sales. The results of our studies confirm the physiological and psychological effects of subliminal stimulation that have already been reported in the specialized literature. Our research on smoking prevention led us to study the sophisticated advertising strategies used by the tobacco industry. We have shown that revealing the subliminal stimuli-at least the visual ones-is extremely useful for teenagers. The enlightened teenager becomes able to recognize the subliminal manipulation concealed in advertising, and the risk of becoming its victim. Such educational efforts have their merits, particularly at the school level. PMID- 9216803 TI - An epidemiological study in an Anatolian village in Turkey environmentally exposed to tremolite asbestos. AB - After several cases of malignant pleural mesothelioma (MPM) were detected in the village of Kureysler in the Kutahya district of western Turkey, an epidemiological study was conducted. A questionnaire was completed by 124 villagers who were older than 20 years and standard posteroanterior chest X-rays were taken. The films were evaluated by three chest physicians. Samples of the white stucco that had been used by almost all villagers for indoor painting for many years were mineralogically examined. Chest X-rays showed that 23 (18%) had pleural plaques and calcifications compatible with asbestos exposure. Male sex and old age were associated with occurrence of pleural plaques. An analysis of white stucco samples revealed tremolite asbestos. In conclusion, tremolite fibers might be the cause of the high incidence of pleural plaques and MPM cases in the village of Kureysler. PMID- 9216804 TI - Environmental fibrous zeolite (erionite) exposure and malignant tumors other than mesothelioma. AB - We studied the mortality in three villages in the Cappadocian region of Central Anatolia, Karain, Tuzkoy, and Sarihidir, which were exposed to fibrous zeolite (erionite), a known carcinogen more potent than the amphibole asbestos. Between 1970 and 1994, there were 305 deaths in Karain, and 177 (58%) were cancer related, including 150 (49.2%) malignant pleural mesothelioma, seven (2.3%) malignant peritoneal mesothelioma, and six (1%) gastroesophageal carcinoma. Four deaths (1.3%) from lung cancer included two nonsmoking females. There were three cases (1%) of leukemia and six of other malignancies (1.9%). Between 1980 and 1994, there were 519 deaths in Tuzkoy (T) and Sarihidir (S) (T = 432, S = 87). Of these, 257 were cancer related, and included 120 cases of malignant pleural mesothelioma and 64 cases of malignant peritoneal mesothelioma. Intraabdominal carcinoma was noted in 29 patients and 14 patients had lung cancer (four of whom were nonsmoking women). There were five cases of gastroesophageal cancer, five deaths due to leukemia, and 16 cases of various malignancies. These mortality figures support the hypothesis that erionite fibers cause cancer other than mesothelioma and lung cancer. Mineralogic analyses of the tissues should be performed to demonstrate this relationship. PMID- 9216805 TI - Malignant peritoneal mesothelioma following asbestos exposure. AB - Clinical, epidemiological, and pathological studies have demonstrated that asbestosis plays a major role in the etiology of mesothelioma. The direct exposure of workers in industrialized countries to asbestos fibers and nonoccupational household contact elevate the risk of malignant mesothelioma. An increased risk has been found in certain geographic areas of Turkey due to the presence of asbestos deposits and the use of the material known as "white soil" as an insulation. We present a malignant mesothelioma case from rural eastern Turkey with a history of asbestos exposure from using "white soil". We review the epidemiological aspects of asbestos as they relate to mesothelioma. PMID- 9216806 TI - Some etio-pathogenetic factors in laryngeal carcinogenesis. AB - Chemical influences, mainly heavy tobacco smoking, chewing snuff, excessive alcohol consumption, and some occupational hazards, are known to be important etiologic factors in laryngeal carcinogenesis. The synergistic or cooperative interaction of human papilloma virus (HPV) infection with these chemical factors are serious considerations in the development of laryngeal carcinoma. With the development during the last decade of Southern blot hybridization and polymerase chain reaction (PCR), extensive and comprehensive studies have been conducted to determine the presence and biological (etiologic) significance of HPV. Developed cancer, as well as juvenile and adult multiple and single papilloma of the larynx, have been the subject of clinical and molecular-pathological investigation. Our previous study showed that cancer may develop on the basis of leukoplakia and adult-onset papilloma. Extensive kilocytes, an indication of HPV infection, can be seen by histological examination in papillomas and carcinoma. Literary data suggest that in laryngeal squamous cell carcinoma, including varicoses carcinoma, HPV 16, HPV 18, and HPV 33 DNA have been detected. Both in juvenile and adult-onset respiratory papillomatosis, patients could have either HPV type 6 or 11 DNA sequences. Molecular biological and PCR studies indicate that HPV may play an etiologic role in the development of human malignancies of the upper aerodigestive tract and uterine (cervical) origin. However, evidence that unequivocally links HPV infection with laryngeal squamous cell carcinoma is still lacking. In laryngeal cancer, p53 abnormalities are related to smoking induced mutagenesis rather than HPV. Studies have postulated an interaction between HPV infection and chemical carcinogens and have concluded that HPV possibly are co-adjuvants during the multistage process of neoplastic transformation. PMID- 9216807 TI - Molecular and epidemiological approaches to the etiology of urinary tract tumors in an area with Balkan endemic nephropathy. AB - Recently, we completed a second biostatistical study of urinary tract tumors (UTT) in areas with Balkan endemic nephropathy (BEN) in the Vratza district, Bulgaria, during the period 1975 to 1991. We confirmed the positive correlation between the incidence of urinary tract tumors (UTT) and BEN demonstrated in our first population-based case control 1977 study. A UTT incidence of 98.9 per 100,000 men and 74.7 per 100,000 women was found in villages most affected by BEN when compared with 11.0 and 6.7 for men and women, respectively, in nonendemic villages. The relative risk (RR) of UTT in BEN villages showed tumors of kidney pelvis and ureters-29 in men and 35 in women and urinary bladder tumors-4 in men and 11 in women. The percentage of food and blood samples containing nephrotoxic and carcinogenic mycotoxin Ochratoxin A (OTA) correlated with the origin of the samples. The most contaminated samples were found in BEN villages and households, and the urinary excretion of OTA was higher in the group of BEN/UTT patients. The UTT DNA's were studied by the 32P-postlabeling method for the presence of OTA-DNA adducts. Some OTA-DNA adducts characteristic for endemic UTT and absent in control nonendemic UTT and nontumorous tissues were described for the first time. PMID- 9216808 TI - The epidemiology of gastrointestinal cancer in Turkey: a review of our accumulated experience. AB - Among all organ cancers, gastrointestinal tract cancers present an interesting pattern in distribution over the world. There are several hundred differences between some incidences of cancer. Probably due to different geographical and climatic differences between western and eastern regions of Turkey, we found varying incidences in esophageal, gastric, and colonic carcinomas. The type of diet, and an excess or lack of some essential nutrients and vitamins are probably the main causes in determining what kind of gastrointestinal tumor might occur. Besides diet, living areas, socioeconomic status, salinity of soil, drinking water and many other factors may play a role. Contrary to the findings of some authors, excessive tea and alcohol consumption has not been found to be a risk factor in our study. PMID- 9216809 TI - Helicobacter pylori and intestinal metaplasia. AB - Gastric carcinoma of the intestinal type is assumed to develop from precancerous gastric lesions. It is now widely accepted that Helicobacter pylori (HP) infection causes chronic gastritis and, after a period of time, intestinal metaplasia (IM). It was suggested that these gastric lesions may evolve into gastric carcinoma after a lengthy latency period. HP seropositivity is high in Turkey at early ages. This may explain the high incidence of gastric carcinoma in this geographic region. In this study, we examine the relationship between HP and IM in endoscopic gastric biopsy specimens. We examined 840 biopsies taken from 210 patients. HP positivity and the presence of IM were examined in these specimens by histopathologic methods. HP positivity was also determined by CLO testing. HP was positive in 156 of the 210 patients examined (74.3%). The distribution of HP seropositivity did not differ between age groups (p > 0.05). IM was present in 101 patients in the entire study group (48%). Among the 156 HP positive patients, the rate of IM was 44.8% (n = 70). The rate of IM among the 54 HP-negative patients was 57.4% (n = 31), which was not statistically significant (p > 0.05). IM positivity has been shown to increase in older age, which was statistically significant (p < 0.001). We were not able to show a relationship between HP seropositivity and IM. Increased HP seropositivity at an early age is a common risk factor in our population. We must consider other factors that may contribute to the increased rate of IM in older age groups. PMID- 9216810 TI - Subtypes of intestinal metaplasia and their relationship to Helicobacter pylori. AB - The presence of incomplete colonic type of intestinal metaplasia (IM) is regarded as a risk factor for gastric carcinoma. In this study, we attempted to classify the subtypes of IM in our patients and examine their relationship to Helicobacter pylori (HP). Gastric biopsies were obtained from 210 patients. The HP positivity and the presence and type of IM were determined by histopathologic methods. HP positivity was also tested by the CLO test. The subtypes of IM were classified according to Ming's classification. IM was present in 101 of 210 patients (48%). Complete type intestinal metaplasia was present in 72 of 101 patients (71.3%), incomplete type IM was seen in 29 of 101 patients (28.7%), and incomplete colonic type (Type IIc) was found in 7 of 101 patients (6.9%). No significant relationship was found between subtypes of IM and HP positivity (p > 0.05). Although our patient group is small, our findings suggest that the carcinogenity of HP is mostly related to its own mutagenic activity as well as the mutagenic activity of the inflammatory cells present in response to HP rather than to its supposed effect on precancerous gastric lesions. PMID- 9216811 TI - Regional epidemiological features of lip, oral cavity, and oropharyngeal cancer. AB - Lip, oral cavity, and oropharyngeal cancer are among the most common forms of the disease in the world. These types of cancer display significant geographic, ethnic, and socioeconomic variations. We examined the cases of cancer of the lip, oral cavity, and oropharynx diagnosed in the Department of Otolaryngology at the University of Uludag School of Medicine during the last 5 years, July 1990 to June 1995, and recorded the epidemiological features of these tumors. The Department of Otolaryngology treated a total of 26,225 in- and outpatients during the 5-year period. 320 of these patients (1.2%) were seen for head and neck cancer. 42 of the 320 patients (13.1%) were diagnosed with cancer of the lip, oral cavity, and oropharrynx. After the larynx, this was the second most frequent location of malignant head and neck tumors. We discovered the following epidemiological and pathological features: (1) The incident rate was highest in patients between 41 and 60 years of age. (2) 70% of the patients were male, and 76% of them had a history of tobacco/alcohol use. (3) Occupation had no apparent relevance (four of the patients were farmers). (4) Approximately one third of the patients had undergone medical therapy prior to diagnosis. (5) One third of the patients had initially seen a dentist for treatment, and approximately half had poor dental and oral hygiene. (6) The most frequent symptom was ulceration. (7) Histopathological examination revealed squamous cell carcinoma in 88% of the cases. (8) The cancer was localized to the lip in 31% of cases, oral cavity, 50%, and oropharynx, 19%. (9) Almost half of the cancer cases were diagnosed in advanced stage (stages III and IV). PMID- 9216812 TI - Topographic distribution of laryngeal cancer. AB - The anatomic distribution of laryngeal cancer (LC) among the compartments of the larynx shows geographic variations. In the U.S., glottic cancers are more frequently seen, whereas most cases in the Mediterranean countries are supraglottic. We reviewed the anatomic sites of involvement in patients with laryngeal cancer seen in our clinic and at eight other university clinics between 1990 and 1994. The majority of cases were supraglottic cancers, accounting for 60% of all laryngeal tumors. PMID- 9216813 TI - Thyroid pathology in residents of the Kiev region, Ukraine, during pre- and post Chernobyl periods. AB - We compared the results of thyroid surgery in residents of the Kiev region in the Ukraine during the 4 years preceding the 1986 Chernobyl nuclear power station (CNPS) accident with the results during the 8 years following the catastrophe. We examined 16,340 thyroid glands, removed from patients during the periods 1982 to 1986 and 1987 to 1995, and found increases in the rates of Hashimoto's disease, follicular adenoma, and thyroid cancer. The increased incidence of papillary microcarcinoma deserves special attention when compared to the rates of clinically manifested forms of thyroid cancer. The resulting data bear evidence to the effect of the environment on the thyroid diseases following the Chernobyl catastrophe. However, the role of certain subjective factors must also be considered. PMID- 9216814 TI - Latent thyroid pathology in residents of Kiev, Ukraine. AB - Thyroid glands from 162 patients were examined for occult thyroid pathology at autopsy. The patients were residents of Kiev, aged 16 to 82 years, without known thyroid disease. Abnormalities were observed in 66 of the 162 cases (40.7%) and included nodular hyperplasia (13.6%), follicular adenoma (4.9%), and thyroiditis (9.5%). Thyroid carcinoma was found in 11.7% of cases (18 papillary microcarcinomas and one follicular oncocytic carcinoma). No cancer was found in the thyroids of patients under 35 years of age. Cancer was predominant in women, but there were no significant differences in the frequency of occult carcinoma with increased age. The neoplasms varied in size from microscopic foci to 9 mm in diameter; most (63%) were over 5 mm. The cancer incidence rate in our study was lower than that found in Finland and Japan, but higher than in most other published series. The absence of a correlation between the rather high frequency of latent thyroid cancer in autopsy data and the relatively low annual incidence of clinically evident thyroid cancer in Ukraine residents may reflect the impact of initiating factors on carcinogenesis of the thyroid. Our results suggest that environmental factors in the area of Ukraine, contaminated by the Chernobyl accident, may play an important role in the etiology of thyroid cancer, but further research is needed. PMID- 9216815 TI - A retrospective analysis of thyroid cancer. AB - We reviewed the results of 2071 thyroidectomies performed during the past 11 years at Uludag University School of Medicine Hospital. Of all the patients 1602 (77.4%) were women and 469 (22.6%) were men (F:M = 3.4:1). Seventy-eight of the thyroid surgery patients (3.77%) had thyroid carcinoma, with a female to male ratio of 2.0:1. The relative risk of thyroid cancer in male versus female patients with thyroid nodules was determined to be 1.75:1. Patient distribution by thyroid carcinoma type was: papillary carcinoma 49%, follicular carcinoma 24%, undifferentiated carcinoma 10%, metastatic carcinoma 10%, and medullary carcinoma 6%. Fine needle aspiration biopsy (FNAB) has been used routinely in our hospital for the last 4 years. During this period, the average number of operations decreased from 201 to 130 per year and the surgical diagnosis of thyroid carcinoma increased from 2.85 to 7.65%. We conclude that papillary carcinoma is relatively less prevalent in our population and that fine needle aspiration biopsy is the preferred method of diagnosing nodular thyroid disease. PMID- 9216816 TI - Breast carcinoma in Pakistani women. AB - In many developed and developing countries including Pakistan, breast cancer is the most frequently diagnosed cancer and the leading cause of cancer death in women. Among 4575 women presenting to the Cancer Research Foundation of Pakistan between 1987 and 1994 with breast lumps, 1201 (26%) were found to have breast cancer. Their ages ranged from 19 to 79 years. The peak incidence was in the 30 to 39 age group. Most patients were multiparous with an average of five children. The size of the tumor was greater than 5 cm in 66% of the cases. Invasive ductal carcinoma was the most common morphological type. According to the Bloom and Richardson grading system, 58% of cases were grade III. Lymph node metastases were present in 73% of the cases. PMID- 9216818 TI - Pediatric malignancies in Bursa, Turkey. AB - Turkey is among the countries affected by ionizing radiation from the Chernobyl accident of April 26, 1986. The northern part of Turkey, where the city of Bursa is located, is presumed to be more influenced by the nuclear catastrophe. The radioactive elements in the atmosphere have been examined at various intervals after May 1, 1986 and barium-140 and lanthanum-140, fission agents of uranium 235, have been found in the atmosphere. Their exact concentration could not be measured. The aim of this report is to review the pediatric malignancies diagnosed in our institution between 1986 and 1995, with a view on any significant increase in the number of these cases. The patients were divided into three groups: acute leukemia patients (101 cases), lymphomas (44 cases), and solid tumors (31 cases). All three groups showed a tendency to increase after 1986; the increase in leukemia cases between 1986 and 1995 was found to be significantly higher (p < 0.001) when compared with the years before 1986. The increase in lymphoma and solid tumor cases after 1986 was not found to be significant (p > 0.05). We cannot rule out environmental causes other than the effects of the Chernobyl accident, and we feel that more intense epidemiological studies should be carried out on this subject in other areas of Turkey. PMID- 9216817 TI - Non-melanoma skin cancer: a case-control study on risk factors and protective measures. AB - There is considerable evidence that exposure to ultraviolet radiation increases the risk of many dermatological conditions, such as non-melanoma skin cancer (NMSC). In order to better understand this relationship, we examined the connection between quantitative measures of individual sun exposure and the risk of NMSC, and the benefits of some protective measures against sunlight, analyzing the differences by sex. A case-control study was conducted in Valencia, Spain during 1990 to 1992 that included 276 cases of histologically confirmed NMSC and 552 control subjects matched by age, sex, and area of residence. Quantitative ultraviolet exposure, phenotypic features, and protective measures from sunlight were estimated by means of a history questionnaire administered by interview. Logistic regression analysis was carried out for each variable and level of quantitative measures. We observed a statistically significant increase in the risk of NMSC proportional to an increase in the hours of occupational exposure to the sun (OR = 1.2, 2.5, and 5.3, respectively). An increased risk of NMSC was observed in men with high nonoccupational exposure (OR = 1.7; p < 0.05 in open air activities in the sun, OR = 2.1; p < 0.05 in the sun while on vacation). In women, we found instances of OR > 1, but without significance (p > 0.05). Wearing a hat at work appeared to be an important protective measure for men. Light phenotypic features predominated in our study, especially in women, and seems to be the major risk factor involved. It seems reasonable to presume that differences between the sexes are basically sociocultural, (i.e., different work activities and different use of leisure time. PMID- 9216819 TI - Multiple myeloma in the region of Bursa, Turkey: a retrospective analysis. AB - We evaluated the clinical and laboratory features of multiple myeloma in our patients and reviewed the factors that affected their survival. The study included 36 patients (12 women and 24 men) with multiple myeloma whom we followed up until death between October 1978 and June 1995. The age range was 34 to 75 years (mean age, 53.9). The chief complaints on admission were lumbar pain and pain in the extremities (77.8%) and generalized weakness (61.1%). The most common laboratory findings were severe anemia (hemoglobin < 8.0 g/dl) (50%), elevated erythrocyte sedimentation rate (75%), monoclonal spike in the serum protein electrophoresis (44.4%), and lytic skull lesions (72.2%). Twenty-three (64%) patients had a monoclonal IgG, 9 (25%) had IgA, 1 had IgD, 2 had light chain disease, and 1 was nonsecretory. Localized plasmacytoma was detected in 4 patients and 4 patients had amyloidosis in rectal and gingival biopsies. According to the Durie-Salmon staging system, 2 patients were in stage 1, 8 were in stage 2, and 26 were in stage 3. The mean survival was 31.4 +/- 4.3 months (range: 1 to 96). The 5-year survival rate was 11%. Sex, age at diagnosis, stage of the disease, hemoglobin level, platelet count, level of serum calcium, creatinine, serum paraprotein, and type of paraproteinemia were tested as prognostic parameters. We could not establish a statistically meaningful effect of these parameters on survival time. The first and second most common causes of death were renal failure and infection, respectively. PMID- 9216820 TI - Paraneoplastic neurological syndrome in systemic cancer. AB - Paraneoplastic syndrome refers to a group of disorders caused by or associated with cancers that are not direct effects of the primary tumor mass or a metastasis to the involved organs. Neurologically, the phrase describes a group of disorders that are diagnosed with increasing frequency in cancer patients. In this study, we investigated 36 patients with malignant diseases and various neurological paraneoplastic syndromes. Lung cancer is the most frequent malignancy associated with neurological paraneoplastic syndromes, and polyneuropathy is the most important manifestation among them. PMID- 9216821 TI - Second primary cancer due to radiotherapy and chemotherapy. AB - Cancer patients are treated successfully with chemotherapy, radiotherapy, or a combination of both. However, many agents used in cancer chemotherapy as well as ionizing radiation are known carcinogens. The long survival of cancer patients treated successfully for their primary cancer made possible the observations of late effects of radiotherapy and chemotherapy and, in particular, the occurrence of second primary cancers. In this report we review the cases of five patients with second primary malignancies and wish to emphasize the importance of a thorough follow-up of patients treated successfully for and possibly cured of a primary cancer. PMID- 9216822 TI - The usefulness of serum levels of CEA, CA 50, and ferritin in the management of renal cell cancer. AB - The aim of this study is to show the usefulness of serum CEA, CA 50, and ferritin levels in the treatment of patients with kidney cancer. Serum CEA and ferritin were determined by EIA assay using Hoffman-la Roche (Austria) kits and serum CA 50 by fluorometric assay using Delfia Pharmacia LKB (Sweden) kits. Cut-off values were 3 ng/mL for CEA, 17 U/mL for CA 50, and 180 ng/mL for ferritin. Increases in the serum levels of CA 50 and ferritin were found to herald the postoperative recurrence of kidney cancer. PMID- 9216823 TI - A comparison of the clinical usefulness of CA 19-9 and CA 50 in the diagnosis and monitoring of gastrointestinal cancers. AB - We compared serum levels of CA 19-9 and CA 50 in 108 patients with malignant neoplasms of the stomach, pancreas, liver, and colon with the serum levels in 60 patients with benign gastrointestinal diseases, and 10 healthy subjects. Increased serum levels of CA 19-9 and CA 50 were found in 51.8 and 62% of the cancer patients, respectively. The results of CA 19-9 and CA 50 assays in the nonneoplastic group showed less specificity. False positive results were noted in 11.7% of CA 19-9 tests and in 31.6% of CA 50 tests. We concluded that in gastrointestinal cancer, the CA 19-9 test should be performed initially. CA 50 determination can be useful, but the lower specificity of the test should be taken into consideration. CA 50 should be recommended only for postoperative monitoring, especially in patients with normal CA 19-9 serum levels. PMID- 9216824 TI - Therapeutic hyperthermia in cancer and AIDS: an updated survey. AB - The aim of this paper is to update with personal contributions the progress thus far accomplished in the clinical application of hyperthermia (HT) in cancer and chronic infectious diseases. The HT treatment has been successfully developed since the 1970s in cancer patients in whom it showed positive results consisting of complete or partial clinical remissions. Its rationale was based on the fact that core temperatures of > or = 42 degrees C induce cytotoxic effects that are higher in malignant cells than in normal cells. HT could be applied by different methods according to type, stage, and localization of the malignancies. Thus, systemic whole-body HT (WBH), through invasive or noninvasive techniques, was first used in disseminated cancers; local perfusion, infusion, and interstitial HTs have been applied in limb, skin, subcutaneous, or intracavitary tumors. The observation of a macrophagic lysosomal exocytosis and subsequent cancer cell death induced by HT, suggested that its mechanism of action involves an immune reaction. This suggested the possibility of associating HT with cytotoxic agents, antibiotics, antiviral drugs, and antioxidants, including beta-carotene (BC). The association of HT with BC at high doses are synergistic in patients with AIDS related complex (ARC) and improve its symptoms, preventing the progress of the disease into the severe stage of AIDS; the same synergism helped also to increase the survival time in patients with severe AIDS. PMID- 9216825 TI - The scientific platform and activity of the Ukrainian INEC Section. AB - The Ukrainian INEC Section is the most recently established chapter of the European Institute of Ecology and Cancer (INEC). We herewith review its organization and goals, membership, scientific platform, and areas of scientific activity. PMID- 9216826 TI - Exceptionally potent antispermatogenic compounds from 8-halogenation of (4aRS,5SR,9bRS)-hexahydroindeno-[1,2-c]pyridines. PMID- 9216827 TI - Inhibition of human telomerase by a G-quadruplex-interactive compound. PMID- 9216828 TI - (S)-(+)-4-[7-(2,2-dimethyl-1-oxopropoxy)-4-methyl-2-[4-[2-(1-piperidinyl) ethoxy]phenyl]-2H-1-benzopyran-3-yl]-phenyl 2,2-dimethylpropanoate (EM-800): a highly potent, specific, and orally active nonsteroidal antiestrogen. PMID- 9216829 TI - (1 alpha, 2 beta, 3 beta, 4 alpha)-1,2-bis[N-propyl-N-(4-phenoxybenzyl) amino]carbonyl]cyclobutane-3,4-dicarboxylic acid (A-87049): a novel potent squalene synthase inhibitor. PMID- 9216830 TI - 5,6-Cis-penems: broad-spectrum anti-methicillin-resistant Staphylococcus aureus beta-lactam antibiotics. AB - 5,6-cis-Penem derivatives have been synthesized and evaluated as anti-MRSA antibiotics. The cis-penems 5 and 6 showed potent activities against not only MRSA but also a wide variety of bacteria including beta-lactamase-producing microorganisms. These compounds were designed to have high affinity to the penicillin-binding protein 2a of MRSA and to form stable acyl intermediates with beta-lactamases by blocking the deacylating water molecule. PMID- 9216831 TI - Discovery of prototype peptidomimetic agonists at the human melanocortin receptors MC1R and MC4R. AB - [Nle4, DPhe7]-alpha-MSH (NDP-MSH), a highly potent analogue of alpha-melanocyte stimulating hormone (alpha-MSH), possesses nanomolar efficacies at all the melanocortin receptor subtypes except the MC2R. Evaluation of the melanocortin "message" sequence of [Nle4, DPhe7]-alpha-MSH was performed on the human melanocortin receptor subtypes designated hMC1, hMC3R, hMC4R, and hMC5R. Tetrapeptides and tripeptides were stereochemically modified to explore topochemical preferences at these receptors and to identify lead peptides possessing agonist activity and subtype selectivity. Four peptides were discovered to only bind to the hMC1 and hMC4 receptor subtypes. The tetrapeptide Ac-His-DPhe-Arg-Trp-NH2 (1) possessed 0.6 microM binding affinity at the hMC1R, 1.2 microM binding affinity at the hMC4R, and agonist activity at both receptors. The tripeptides Ac-DPhe-Arg-Trp-NH2 (6) and Ac-DPhe-Arg-DTrp-NH2 (7) possessed 2.0 and 9.1 microM binding affinities, respectively, only at the hMC4R, and both compounds effected agonist activity. The tetrapeptide Ac-His-Phe-Arg-DTrp-NH2 (4) possessed 6.3 microM affinity and full agonist activity at the hMC1R, while only binding 7% at the hMC3R, 36% at the hMC4R, and 11% at the hMC5R at a maximal concentration of 10 microM. These data demonstrate that the His-Phe-Arg-Trp message sequence of the melanocortin peptides does not bind and stimulate each melanocortin receptor in a similar fashion, as previously hypothesized. Additionally, this study identified the simplest structural agonists for the hMC1R and hMC4R receptors reported to date. PMID- 9216833 TI - Structural model of a cyclic dynorphin A analog bound to dodecylphosphocholine micelles by NMR and restrained molecular dynamics. AB - The compound c[Cys5,11]dynorphin A-(1-11)-NH2, 1, is a cyclic dynorphin A analog that shows similar selectivity and potency at the kappa-opioid receptor when compared to the native form of the peptide in central nervous system assays. Previous molecular mechanics calculations have shown that the ring portion of the isoform that is trans about the Arg9-Pro10 omega bond contains either a beta-turn from residues Arg6 to Arg9 or an alpha-helical conformation. Our results from solution state NMR indicate that the compound exhibits cis-trans isomerism about the Arg9-Pro10 omega bond in both aqueous solution and when bound to dodecylphosphocholine micelles. Restrained molecular dynamics calculations show that the cis isoform of the peptide contains a type III beta-turn from residues Arg7 to Pro10. Similar calculations on the trans isoform show it to contain a beta-turn from residues Cys5 and Arg8. In this report we describe the generation of three-dimensional models from NMR data for the ring portions of both the cis and trans isoforms of 1 bound to dodecylphosphocholine micelles. Comparison with other dynorphin A structural information indicates that both the cis and trans isoforms of the peptide may be active as kappa-opioid agonists. PMID- 9216834 TI - Substituted 3-(phenylsulfonyl)-1-phenylimidazolidine-2,4-dione derivatives as novel nonpeptide inhibitors of human heart chymase. AB - A series of 3-(phenylsulfonyl)-1-phenylimidazolidine-2,4-dione derivatives have been synthesized and evaluated for their ability to selectively inhibit human heart chymase. The structure-activity relationship studies on these compounds gave the following results. The 1-phenyl moiety participates in a hydrophobic interaction where an optimum size is required. At this position, 3,4 dimethylphenyl is the best moiety for inhibiting chymase and showed high selectivity compared with chymotrypsin and cathepsin G. A 3-phenylsulfonyl moiety substituted with hydrogen-bond acceptors such as nitrile and methoxycarbonyl enhances its activity. Molecular-modeling studies on the interaction of 3-[(4 chlorophenyl)sulfonyl]-1-(4-chlorophenyl)-imidazolidine-2,4-dione (29) with the active site of human heart chymase suggested that the 1-phenyl moiety interacts with the hydrophobic P1 pocket, the 3-phenylsulfonyl moiety resides in the S1' S2' subsites, and the 4-carbonyl of the imidazolidine ring and sulfonyl group interact with the oxyanion hole and the His-45 side chain of chymase, respectively. The complex model is consistent with the structure-activity relationships. PMID- 9216832 TI - Further definition of the D1 dopamine receptor pharmacophore: synthesis of trans 6,6a,7,8,9,13b-hexahydro-5H-benzo[d]naphth[2,1-b]azepines as rigid analogues of beta-phenyldopamine. AB - In an effort to define further the active geometry of the beta-phenyldopamine pharmacophore of certain dopamine D1 agonists, the title compounds have been synthesized as conformationally restricted homologues of the potent benzophenanthridine dopamine D1 agonist dihydrexidine 4a. The dihydroxy secondary amine 5b was evaluated as a potential agonist, whereas the N-methyl compounds 5a and 5c were hypothesized to be antagonists. Surprisingly, none of the three compounds had high affinity for dopamine D1 or D2 receptors. A comparison of the low-energy conformations of these molecules shows that the pendant phenyl ring of 5b is twisted about 28 degrees relative to that of the corresponding ring of 4a. Further, the additional methylene used to expand the C ring of 5b projects toward the alpha face of the molecule, perhaps suggesting that steric protrusion in this region of the molecule is not tolerated. Finally, the phenethylamine fragment incorporated into these molecules deviates about 30 degrees from the antiperiplanar conformation postulated to be necessary for agonist activity. On the other hand, the potential antagonist molecules 5a and 5c were compared with the dopamine D1 antagonist SCH 39166 2. The conformations of the former two structures differ quite dramatically from that of 2. The most notable differences lie in the relative orientations of the pendant phenyl rings in the two series, as well as the fact that the ethylamine fragment in 2 approximates a gauche conformation, while the comparable orientation in 5a and 5c more nearly approaches an antiperiplanar conformation. These findings will be used to refine further the model of the dopamine D1 agonist receptor that we have previously developed. PMID- 9216835 TI - Potent HIV protease inhibitors containing a novel (hydroxyethyl)amide isostere. AB - A series of HIV protease inhibitors containing a novel (hydroxyethyl)amidosuccinoyl core has been synthesized. These peptidomimetic structures inhibit viral protease activity at low nanomolar concentrations (IC50 < 10 nM for HIV-1 protease). The inhibition constant (Ki) for inhibitor 19 was determined to be 7.5 pM against HIV-1 and 1.2 nM against HIV-2 proteases, respectively. Several compounds (19-24) inhibited HIV-1 replication in cell culture assays with 50% effective concentrations (EC50) = 3.7-35 nM. This series of inhibitors was found to exhibit poor bioavailability (< 10%) in the rat, following oral administration. The synthesis and biological properties of these compounds are discussed. In addition, an X-ray structure of one of these inhibitors (23) in complex with HIV-2 protease provides insight into the binding mode of this novel class of HIV protease inhibitors. PMID- 9216836 TI - A facile, alternative synthesis of 4'-thioarabinonucleosides and their biological activities. AB - 4'-Thioarabinonucleosides, which are potential antiviral agents, were synthesized from D-glucose. 1,4-Anhydro-4-thioarabitol (8), which can be derived from diacetone glucose in nine steps, was subjected to Pummerer rearrangement after protection of the hydroxyl groups to give 1-O-acetyl-4-thioarabinose (11), which was condensed with nucleobases to give 4'-thioarabinonucleosides. The 5 substituted-4'-thioaraU (6a-e) derivatives showed anti-HSV-1 activity (ED50 = 0.43-3.50 micrograms/mL). 4'-ThioaraG (6h) and 2,6-diaminopurine 4' thioarabinonucleoside (4'-thioaraDAP, 6g) showed antiviral activity against several herpes viruses and were particularly potent against human cytomegalovirus (0.010 and 0.022 microgram/mL, respectively). PMID- 9216837 TI - Synthesis and cellular uptake of 2'-substituted analogues of (E)-5-(2 [125I]iodovinyl)-2'-deoxyuridine in tumor cells transduced with the herpes simplex type-1 thymidine kinase gene. Evaluation as probes for monitoring gene therapy. AB - A useful synthetic methodology was developed to synthesize and radiolabel a series of (E)-5-(2-[125I]iodovinyl)uracil nucleoside substrates for herpes simplex virus type-1 thymidine kinase (HSV-1 TK). (E)-5-(2-[125I]Iodovinyl)-2' deoxyuridine ([125I]IVDU, 10), (E)-5-(2-[125I]iodovinyl)-2'-fluoro-2' deoxyuridine ([125I]IVFRU, 11), (E)-5-(2-[125I]iodovinyl)-2'-fluoro-2' deoxyarabinouridine ([125I]IVFAU, 12), and (E)-5-(2 [125I]iodovinyl)arabinouridine ([125I]IVAU, 13) were synthesized in 63-83% radiochemical yield by reaction of the unprotected (E)-5-(2 (trimethylsilyl)vinyl) precursors (6-9) with [125I]ICl. Cellular uptake of these labeled compounds (10-13) was evaluated in vitro. All compounds showed minimal uptake in the KBALB cell line. However, increased uptake was observed for all compounds in KBALB-STK cells which are transduced with a replication incompetent Moloney murine leukemia virus vector encoding the HSV-1 TK gene. The results indicate that uptake of these compounds in KBALB-STK cells is variable and highly dependent on the nature of the sugar 2'-substituent. When a fluoro (12) or a hydroxy (13) substituent is present in the arabinofuranosyl (up) configuration at the 2'-position, there is diminished cellular uptake in KBALB-STK cells relative to hydrogen (10) or fluorine (11) in the ribofuranosyl (down) configuration at the 2'-position. Our results indicate that radiolabeled IVFRU (11) is most promising for further in vivo studies. PMID- 9216838 TI - Phosphodiester amidates of unsaturated nucleoside analogues: synthesis and anti HIV activity. AB - The effect of introduction of a lipophilic phosphodiester amidate moiety on the HIV activity of inactive unsaturated nucleoside analogues was investigated. Phosphodiester alaninates 5a, 5b, and 6 derived from unsaturated nucleoside analogues 3b, 3c, and 4a were synthesized and investigated as inhibitors of cytopathic effect and replication of HIV-1 in ATH-8 cells. Compound 5a is an inhibitor of HIV-1 whereas analogue 6 is inactive with cytotoxicity appearing above 10 microM and 5b is both inactive and nontoxic. Alkaline or enzymic hydrolysis of 5a gave phosphomonoester alaninate 14, a putative product of intracellular metabolism. Compound 14 as well as adenallene derivative 15c were devoid of anti-HIV activity, and they also failed to inhibit HIV reverse transcriptase. A new regioselective method for preparation of (Z)-4-(benzoyloxy) 1-hydroxy-2-butene, 7, a key intermediate for the synthesis of unsaturated nucleoside analogues of cis configuration such as 3a, 3b, and 3c, is also described. PMID- 9216839 TI - Potent tetracyclic guanine inhibitors of PDE1 and PDE5 cyclic guanosine monophosphate phosphodiesterases with oral antihypertensive activity. AB - Tetracyclic guanines have been shown to be potent and selective inhibitors of the cGMP-hydrolyzing enzymes PDE1 and PDE5. In general, these compounds are inactive or only weakly active as inhibitors of PDE3, which is a major isozyme involved in cAMP hydrolysis. Structure-activity relationships are developed at N-1, C-2, N-3, and N-5 on the core nucleus. Compound 31, with an IC50 of 70 pM, is the most potent inhibitor of PDE1, while 50, with an IC50 of 4 nM, is the most potent inhibitor of PDE5. Compounds 20, 22, 30, and 50 are potent dual inhibitors with IC50 values below 30 nM for both PDE1 and PDE5. Compounds 12, 20, and 28 reduced blood pressure by more than 45 mmHg when administered orally at 10 mg/kg to the spontaneously hypertensive rat (SHR). PMID- 9216840 TI - Iron-mediated generation of the neurotoxin 6-hydroxydopamine quinone by reaction of fatty acid hydroperoxides with dopamine: a possible contributory mechanism for neuronal degeneration in Parkinson's disease. AB - Exposure of dopamine to an excess of linoleic acid 13-hydroperoxide (13 hydroperoxyoctadecadienoic acid) in the presence of ferrous ions in Tris buffer, pH 7.4, resulted in a relatively fast, oxygen-independent reaction exhibiting first-order kinetics with respect to both catecholamine and metal concentrations. Product analysis in the early stages revealed the presence of significant amounts of the quinone of the neurotoxin 6-hydroxydopamine, together with some aminochrome and ill-defined melanin-like material. Quinone formation required the presence of iron, either in the ferrous or ferric form, and was unaffected by peroxidase, catalase, and hydroxyl radical scavengers, e.g. mannitol, as well as biologically relevant antioxidants, like ascorbate and glutathione. Hydrogen peroxide proved as effective as linoleic acid hydroperoxide in inducing dopamine oxidation and conversion to 6-hydroxydopamine quinone. Metal chelators, including EDTA and bipyridyl, markedly suppressed quinone formation without, however, inhibiting dopamine oxidation. These and other results are consistent with a hydroxyl radical independent hydroxylation/oxidation mechanism basically different from the Fenton reaction, which involves direct interaction of the peroxide with a dopamine-Fe(III) chelate generated during the process. PMID- 9216841 TI - Steroidal affinity labels of the estrogen receptor. 3. Estradiol 11 beta-n-alkyl derivatives bearing a terminal electrophilic group: antiestrogenic and cytotoxic properties. AB - With the aim of developing a new series of steroidal affinity labels of the estrogen receptor, six electrophilic 11 beta-ethyl (C2), 11 beta-butyl (C4), or 11 beta-decyl (C10) derivatives of estradiol bearing an 11 beta-terminal electrophilic functionality, i.e. bromine (C4), (methylsulfonyl)oxy (C2 and C4), bromoacetamido (C2 and C4), and (p-tolylsulfonyl)oxy (C10), were synthesized. The range of their affinity constants for binding the estrogen receptor was 0.4-37% that of estradiol; the order of increasing affinity (i) relative to the 11 beta alkyl arm was ethyl < butyl and (ii) relative to the electrophilic functionality was bromoacetamido < bromine < (methylsulfonyl)oxy. Regardless of the conditions used, including prolonged exposure of the receptor to various pH levels (7-9) and temperatures (0-25 degrees C), the extent of receptor affinity labeling by the 11 beta-ethyl and 11 beta-butyl compounds, if any, was under 10%. This was in sharp contrast to results obtained using 11 beta-((tosyloxy)decyl)estradiol which labeled from 60% to 90% of the receptor hormone-binding sites with an EC50 of approximately 10 nM. Estrogenic and antiestrogenic activities of the compounds were determined using the MVLN cell line, which was established from the estrogen responsive mammary tumor MCF-7 cells by stable transfection of a recombinant estrogen-responsive luciferase gene. The two 11 beta-ethyl compounds were mainly estrogenic, whereas the three 11 beta-butyl and the 11 beta-decyl compounds essentially showed antiestrogenic activity. The fact that the chemical reactivities of 11 beta-ethyl and 11 beta-butyl compounds were not compromised by interaction with the estrogen receptor made the synthesized high-affinity compounds potential cytotoxic agents which might be able to exert either (i) a specific action on estrogen-regulated genes or (ii) a more general action in estrogen-target cells. Therefore the ability of the compounds (1) to irreversibly abolish estrogen-dependent expression of the luciferase gene and (2) to affect the proliferation of MVLN cells were determined. All electrophiles were able to irreversibly suppress expression of the luciferase gene; the antiestrogenic electrophiles were more potent than the estrogenic ones but less efficient than 4 hydroxytamoxifen, a classical and chemically inert triphenylethylene antiestrogen. Only the antiestrogenic electrophiles decreased cell proliferation; however, they were less potent than 4-hydroxytamoxifen. In conclusion, the synthesized electrophilic estradiol 11 beta-ethyl and 11 beta-butyl derivatives (i) were not efficient affinity labels of the estrogen receptor and (ii) did not display significant cytotoxicity in estrogen-sensitive mammary tumor cells. However, since these derivatives displayed high affinity for the estrogen receptor, they could be used to prepare potential cytotoxic agents which might be selective for tumors affecting estrogen-target tissues, by coupling them with a toxic moiety. PMID- 9216842 TI - Design of a potent combined pseudopeptide endothelin-A/endothelin-B receptor antagonist, Ac-DBhg16-Leu-Asp-Ile-[NMe]Ile-Trp21 (PD 156252): examination of its pharmacokinetic and spectral properties. AB - The endothelins (ETs) are a family of bicyclic 21-amino acid peptides that are potent and prolonged vasoconstrictors. It has been shown that highly potent combined ETA/ETB receptor antagonists can be developed from the C-terminal hexapeptide of ET (His16-Leu17-Asp18-Ile19-Ile20-Trp21), such as Ac-(D)Dip16-Leu Asp-Ile-Ile-Trp21 (PD 142893) and Ac-DBhg16-Leu-Asp-Ile-Ile-Trp21 (PD 145065). However, these compounds are relatively unstable to enzymatic proteolysis as determined in an in vitro rat intestinal perfusate assay. This instability is thought to be due to carboxypeptidase activity. In fact, incubation of PD 145065 with carboxypeptidase inhibitors greatly increased its half-life in rat intestinal perfusate. By performing a reduced amide bond and N-methyl amino acid scan, it was discovered that N-methylation of Ile-20 resulted in a compound (Ac DBhg16-Leu-Asp-Ile-[NMe]Ile-Trp21, PD 156252) that retained full receptor affinity at both endothelin receptor subtypes along with enhanced proteolytic stability and cellular permeability. Interestingly, N-methylation of this bond allows the cis configuration to be readily accessible which greatly alters the preferred structure of the entire molecule and may be responsible for the observed enhanced metabolic stability. PMID- 9216843 TI - Lanthionine-somatostatin analogs: synthesis, characterization, biological activity, and enzymatic stability studies. AB - A series of cyclic somatostatin analogs containing a lanthionine bridge have been subjected to studies of structure-activity relationships. A direct synthesis of the thioether bridged analog (1) of sandostatin (SMS 201,995) and several lanthionine hexa-, hepta-, and octapeptides was carried out by using the method of cyclization on an oxime resin (PCOR) followed by condensation reactions in solution. The structures of the target peptides were analyzed by liquid secondary ion mass spectrometry (LSIMS) and subjected to high-energy collision-induced dissociation (CID) studies after opening of the peptide ring by proteolytic cleavage. The biological activities of these compounds have been evaluated by assaying their inhibitory potencies for the release of growth hormone (GH) from primary cultures of rat anterior pituitary cells, as well as by their binding affinities to cloned somatostatin receptors (SSTR1-5). The structural modification of sandostatin by introducing a lanthionine bridge resulted in a significantly increased receptor binding selectivity. The lanthionine octapeptide with C-terminal Thr-ol (1) showed similar high affinity for rat SSTR5 compared to somatostatin[1-14] and sandostatin. However, it exhibits about 50 times weaker binding affinity for mSSTR2b than sandostatin. Similarly, the lanthionine octapeptide with the C-terminal Thr-NH2 residue (2) has higher affinity for rSSTR5 than for mSSTR2B. Both peptides (compounds 1 and 2) have much lower potencies for inhibition of growth hormone secretion than sandostatin. This is consistent with their affinities to SSTR2, the receptor which is believed to be linked to the inhibition of growth hormone release by somatostatin and its analogs. The metabolic stability of lanthionine-sandostatin and sandostatin have been studied in rat brain homogenates. Although both compounds have a high stability toward enzymatic degradation, the lanthionine analog has a 2.4 times longer half-life than sandostatin. The main metabolites of both compounds have been isolated and identified by using an in vivo technique (cerebral microdialysis) and mass spectrometry. PMID- 9216845 TI - Conformational dynamics of thyroid hormones by variable temperature nuclear magnetic resonance: the role of side chain rotations and cisoid/transoid interconversions. AB - 1H NMR spectra of the thyroid hormone thyroxine recorded at low temperature and high field show splitting into two peaks of the resonance due to the H2,6 protons of the inner (tyrosyl) ring. A single resonance is observed in 600 MHz spectra at temperatures above 185 K. An analysis of the line shape as a function of temperature shows that the coalescence phenomenon is due to an exchange process with a barrier of 37 kJ mol-1. This is identical to the barrier for coalescence of the H2',6' protons of the outer (phenolic) ring reported previously for the thyroid hormones and their analogues. It is proposed that the separate peaks at low temperature are due to resonances for H2,6 in cisoid and transoid conformers which are populated in approximately equal populations. These two peaks are averaged resonances for the individual H2 and H6 protons. Conversion of cisoid to transoid forms can occur via rotation of either the alanyl side chain or the outer ring, from one face of the inner ring to the other. It is proposed that the latter process is the one responsible for the observed coalescence phenomenon. The barrier to rotation of the alanyl side chain is > or = 37 kJ mol-1, which is significantly larger than has previously been reported for Csp2-Csp3 bonds in other Ph-CH2-X systems. The recent crystal structure of a hormone agonist bound to the ligand-binding domain of the rat thyroid hormone receptor (Wagner et al. Nature 1995, 378, 690-697) shows the transoid form to be the bound conformation. The significant energy barrier to cisoid/transoid interconversion determined in the current study combined with the tight fit of the hormone to its receptor suggests that interconversion between the forms cannot occur at the receptor site but that selection for the preferred bound form occurs from the 50% population of the transoid form in solution. PMID- 9216844 TI - A refined model for the somatostatin pharmacophore: conformational analysis of lanthionine-sandostatin analogs. AB - We report the conformational analysis of a series of analogs of sandostatin (octreotide, D-Phe1-c[Cys2-Phe3-D-Trp4-Lys5-Thr6-Cys 7]-Thr8-ol) using 1H NMR spectroscopy and molecular modeling. Two active compounds in which the disulfide group is replaced by a monosulfide (lanthionine) bridge (D-Phe1-c[AlaL2-Phe3-D Trp4-Lys5-Thr6-A laL7]-Thr8-ol and D-Phe1-c[AlaL2-Phe3-D-Trp4-Lys5-Thr6-Al aL7] Thr8-NH2, where AlaL denotes each of the lanthionine amino acid ends linked by the monosulfide bridge) show different mSSTR2b/rSSTR5 receptor selectivities as compared to sandostatin. These new results have enabled us to reveal features of the somatostatin pharmacophore common to the model previously proposed in our laboratory on the basis of main chain and side chain chiral methylation studies. In addition, our studies provide new insight into the role of the disulfide bridge and of Thr8 in binding potency. We also show that the lanthionine group is a good mimetic of beta-VI turns and can be incorporated in sandostatin analogs maintaining the essential secondary structural features of sandostatin. These results facilitate the design of new sandostatin peptidomimetics. PMID- 9216846 TI - Antitumor agents. 174. 2',3',4',5,6,7-Substituted 2-phenyl-1,8-naphthyridin-4 ones: their synthesis, cytotoxicity, and inhibition of tubulin polymerization. AB - Two series of 2',3',4',5,6,7-substituted 2-phenyl-1,8-naphthyridin-4-ones and 2 phenylpyrido[1,2-alpha]pyrimidin-4-ones have been synthesized and evaluated as cytotoxic compounds and as inhibitors of tubulin polymerization. Most 2-phenyl 1,8-naphthyridin-4-ones showed potent cytotoxic and antitubulin activities, whereas 2-phenylpyrido[1,2-alpha]pyrimidin-4-ones showed no activity in either assay. In general, a good correlation was found between cytotoxicity and inhibition of tubulin polymerization in the 2-phenyl-1,8-naphthyridin-4-one series. The 2-phenyl-1,8-naphthyridin-4-ones (44-49) with a methoxy group at the 3'-position showed potent cytotoxicity against most tumor cell lines with GI50 values in the low micromolar to nanomolar concentration range in the National Cancer Institute's 60 human tumor cell line in vitro screen. Introduction of substituents (e.g. F, Cl, CH3, and OCH3) at the 4'-position led to compounds with reduced or little activity and substitution at the 2'-position resulted in inactive compounds. The effects of various A-ring substitutions on activity depend on the substitution in ring C. Compounds 44-50 were potent inhibitors of tubulin polymerization, with activity nearly comparable to that of the potent antimitotic natural products colchicine, podophyllotoxin, and combretastatin A-4. Compounds 44-49 also inhibited the binding of radiolabeled colchicine to tubulin, but the inhibition was less potent than that obtained with the natural products. Further investigation is underway to determine if substitution at the 3'-position and multisubstitutions in ring C will result in compounds with increased activity. PMID- 9216847 TI - Synthesis and cytotoxic evaluation of substituted sulfonyl-N-hydroxyguanidine derivatives as potential antitumor agents. AB - A series of sulfonyl-N-hydroxyguanidine derivatives was designed and synthesized for cytotoxic evaluation as potential anticancer agents on the basis of the lead compound LY-181984. Replacement of the ureido moiety of the lead compound with hydroxyguanidine provided a stable cytotoxic agent. The conformation of sulfonyl N-hydroxyguanidine derivatives, such as N-(4-chlorophenyl)-N' [(benzo[2,1,3]thiadiazol-4-yl)sulfonyl]-N"- hydroxyguanidine (4g), investigated utilizing HMBC NMR, theoretical calculations, and X-ray crystallography, indicated stacking of the two aromatic rings. The derivatives were evaluated for in vitro cytoxicity against five human tumor cell lines, including HepG2, TSGH 8302, COLO 205, KB, and MOLT-4. The cytotoxic activities of the derived compounds against the human tumor cell lines were equal to or greater than that of the lead compound. N-(4-Chlorophenyl)-N'-[[3,5-dichloro-4-(4-nitrophenoxy)phenyl]sulfonyl] N"- hydroxyguanidine (4n) and N-(4-chlorophenyl)-N'-[[3,5-dichloro-4-(2-chloro-4 nitrophenoxy)phenyl] sulfonyl]-N"-hydroxyguanidine (4o) exhibited the greatest growth inhibition of solid tumor cell lines. Compound 4o was found to possess antitumor activity against murine K1735/M2 melanoma xenografts. PMID- 9216848 TI - The road to the cure of acute lymphoblastic leukemia: a personal perspective. AB - Acute lymphoblastic leukemia was the first syngeneic systemic malignancy to be cured by chemotherapy (1962). The personal description of the key players and the major innovations are presented by one of those participating in this achievement. The principles developed over the subsequent 30 years have found wide application to the therapy of other systemic malignancies. PMID- 9216849 TI - Dacarbazine and interferon alpha for stage IV malignant melanoma. AB - Based on encouraging reports of improved response rates with the use of dacarbazine (DTIC) in combination with recombinant interferon alpha-2a (rIFN alpha-2a) in patients with metastatic malignant melanoma, we conducted a phase II study to determine the efficacy and feasibility of this treatment regimen. 31 patients were treated with an induction dose of rIFN-alpha-2a at 15 MIU/ m2 intravenously (i.v.) daily for 5 days per week for 3 consecutive weeks followed by a continuous maintenance dose of 10 MIU/m2 subcutaneously (s.c.) given 3 days per week; starting on day 22, in conjunction with rIFN-alpha-2a s.c., DTIC was started at a dose of 200 mg/m2 i.v. for 5 continuous days completing a 28-day cycle. Therapy was continued until progression was evidenced. Of the 29 evaluable patients, 7 (24.1%) achieved an objective response (complete plus partial remission) with the highest responses occurring in those patients assessed with pulmonary metastases. The median duration to treatment failure was 2.6 months, while the median survival was 6.9 months. Our data reveal that using rIFN-alpha 2a plus DTIC in combination does not yield better results than those achieved when using DTIC alone. However, 3 of the 7 responders experienced long-term survival ranging up to 42 months. Whether this benefit is achieved by the addition of rIFN-alpha-2a can only be answered by large randomized clinical trials. Conflicting results with some of the current literature are discussed. PMID- 9216850 TI - Weekly 24-hour infusion of high-dose 5-fluorouracil and leucovorin in the treatment of advanced gastric cancers. An effective and low-toxic regimen for patients with poor general condition. AB - Systemic chemotherapy for advanced gastric cancer is frequently associated with significant treatment-related toxicity, which is particularly serve in patients presenting with a poor general condition. A search for effective and low-toxic regimens for this group of patients is mandatory. A weekly 24-hour infusion of high-dose 5-fluorouracil (5-FU) and leucovorin (HDFL) has previously been demonstrated to be an effective treatment for advanced colorectal cancer with minimal toxicity. In the past 3 years, this regimen has been tested at our institutes in patients with advanced gastric cancer, the general condition of whom had made the use of intensive combination chemotherapy impossible. The regimen consisted of a weekly 24-hour infusion of 2,600 mg/m2 of 5-FU and 300 mg/m2 of leucovorin. From August 1992 to December 1995, 34 patients had been treated with this regimen for a total of 488 courses (average: 14.4 per patient). Hematological toxicity of this regimen was minimal, with grade 3 or 4 leukopenia developing in only 1 (2.9%) patient. Other nonhematological toxicities were also negligible except a reversible neurotoxicity which developed in 2 patients. Twenty-five patients were eligible for response analysis. One complete response, 11 partial responses, 5 stable diseases, and 8 progressive diseases were observed. The response rate was 48% (32-72%, 95% CI). The median overall survival (OS) of the whole group was 7 months (range: 1-18+). The median OS and time to progression of the responders were 8.5 months (range: 2-18) and 5 months (range: 2-10+), respectively. The palliative effect was satisfactory with the Karnofsky performance status of the responders improving from a median of 50% (range: 20 90%) to 70% (range: 50-100%). Our retrospective data suggested that HDFL is an effective and low-toxic palliative treatment even in patients with a very poor general condition. We advocated that this regimen should be further tested in ordinary patients with advanced gastric cancer. PMID- 9216851 TI - A randomized trial of cisplatin, vinblastine, and bleomycin versus cyclophosphamide, aclacinomycin, and cisplatin in epithelial ovarian cancer. AB - After primary cytoreductive surgery, a randomized clinical trial was conducted in women with epithelial ovarian cancer to compare the impact on survival between PVB chemotherapy, consisting of cisplatin, vinblastine and bleomycin, and CAP chemotherapy, consisting of cyclophosphamide, aclacinomycin and cisplatin. There were 148 evaluable patients. One hundred and five patients with stage II, III and IV were analyzed in this study, 49 of them received PVB chemotherapy while the remaining 56 patients received CAP chemotherapy. Sixty-four patients fulfilled the criteria for clinical remission set by the Tokai Ovarian Tumor Study Group [Gynecol Oncol 1993;48:342-348]. The remission rate was 73 and 50% in the PVB and CAP groups, respectively, and showed a significant advantage for the PVB group (p = 0.0139). Moreover, the recurrence rate was 44% in the PVB group and 61% in the CAP group after clinical remission, although there was no significant difference between the two groups. The final survival rate was 32% in the PVB group and 24% in the CAP group. There was a significant difference of survival rate between both groups at 24 months (p = 0.0378) and 48 months (p = 0.0450), but finally no significant difference was found at 96 months (p = 0.0660). Compared to the CAP regimen, the PVB combination has a significantly higher efficacy in remission, but there was no significant difference in the long-term survival rate. Furthermore, multivariate analysis demonstrated that the PVB chemotherapy improved the survival, but it was not significant. The authors conclude that PVB chemotherapy may be more effective than CAP chemotherapy for epithelial ovarian cancer. PMID- 9216852 TI - An evaluation of postoperative follow-up tests in colon cancer patients treated for cure. AB - BACKGROUND: Currently patients with colon cancer who are potentially cured by surgery are followed periodically with physical examinations, blood tests and imaging studies to detect tumor recurrence early, on the presumption that intervention can effect outcome. There is little information to indicate whether frequent visits to the doctor's office or frequent testing improves survival or quality of life. METHODS: Ninety-eight patients with resected stage B2, B3 or C (modified Astler-Coller) colon cancer who developed recurrent disease while enrolled in prospective adjuvant trials at Mayo Clinic sponsored by the North Central Cancer Treatment Group were studied to evaluate the utility of follow-up tests to detect the first recurrence of colon cancer and the outcome following various interventions for these recurrences. These patients had a history, physical examination, complete blood count, chemistry panel and chest x-ray approximately every 3-4 months in the 1st year and then every 6-12 months thereafter for a total of 5 years. Bowel evaluation was done at 6 months, 12 months and annually thereafter. In addition, a minority of patients had carcinoembryonic antigen (CEA) testing, and radioisotope liver scans at various intervals. RESULTS: Symptoms signaled the diagnosis of recurrent disease in 55 patients, physical examination in 4 patients, and abnormalities in chest x-ray in 18 patients. An elevated CEA was the initial abnormal test in 5 patients, abnormal liver scans in 5 patients, elevated liver function tests in 6 patients and laparotomy for other reasons in 2 patients. Hemoglobin, barium enema, and fecal blood testing were useful in 1 patient each. Thirty-one percent of recurrences were diagnosed between scheduled visits. In our series, histories, physical examinations, and chest x-rays led to the detection of 79% of the recurrences while liver function tests, liver scans and CEAs led to the detection of 16% of recurrences. Sixteen patients underwent resection for cure for their first recurrence; the diagnosis of recurrence was signaled by symptoms in 6 patients, chest x-ray in 6 patients and abnormal liver function tests, CEA, hemoglobin, and laparotomy for colostomy closure in 1 patient each. CONCLUSIONS: The majority of tumor recurrences were detected by symptoms, physical examinations and chest x-rays. Testing for asymptomatic tumor recurrences during the 1st follow-up year is likely to be much less fruitful for detecting resectable recurrences than testing patients in the 2nd through 4th follow-up years. Patients who had a disease recurrence in the 1st postoperative year were less likely to be candidates for curative intent surgery. Lower tumor grade at initial diagnosis correlated both with likelihood of undergoing secondary surgical resection and the chance of doing well following this. These data may be helpful for defining more appropriate follow-up test for detection of tumor recurrence in patients with resected colon cancer. PMID- 9216854 TI - Phase II study of 3-hour infusion of paclitaxel in patients with previously untreated stage III and IV non-small cell lung cancer. West Japan Lung Cancer Group. AB - Sixty patients with previously untreated non-small cell lung cancer of stages III and IV were treated with a 210 mg/m2 dose of paclitaxel by means of a 3-hour infusion. The objective response rate was 32% (95% confidence interval, 20-45%): 1 complete response and 18 partial responses. The median duration of response was 15 weeks, and the projected median survival duration of all patients was 30 weeks. Grade 3-4 neutropenia occurred in 73% of patients. Other grade 3-4 adverse events included anemia (5%), vomiting/nausea (8%), peripheral edema (2%), alopecia (7%), elevation of AST (2%), peripheral neuropathy (3%), allergic reaction (2%), arthralgia/myalgia (3%), and interstitial pneumonitis (3%). Paclitaxel administered at 210 mg/m2 by means of a 3-hour infusion every 3 weeks demonstrated a notable activity against previously untreated advanced non-small cell lung cancer, with a 32% major response rate. Major toxicity was neutropenia. Hypersensitivity, neurotoxicity, arthralgia/myalgia and cardiac toxicity were mild and easily managed. PMID- 9216853 TI - Weekly CAF chemotherapy for advanced breast cancer patients. AB - In a prospective phase II study, 102 women with advanced breast cancer were treated with low doses of cyclophosphamide, Adriamycin and 5-fluorouracil (CAF) at weekly intervals by intravenous injection. Seventy-five patients were evaluable for treatment response and the overall response rate was 52% (95% confidence interval, 41-63%). Of the evaluable patients, 15% had complete response and 37% had partial response. The median survival after therapy was 15.6 months, the median time to progression was 6.8 months and the median duration of response was 9.1 months. The main toxicities were mild vomiting and moderate myelosuppression. There was only 1 patient who experienced heart failure. Weekly CAF appears to have an efficacy with tolerable side effects comparable to standard CAF with an every-3-week schedule. PMID- 9216855 TI - Loss of the tumor suppressor p53 gene at the liver cirrhosis stage in Japanese patients with hepatocellular carcinoma. AB - The incidence of p53 gene aberrations is reported to be about 20-50% in hepatocellular carcinomas (HCCs). In most cases, HCC is clinically preceded by liver cirrhosis, but the genetic changes in cirrhosis are not known well. Therefore, we studied the loss of heterozygosity (LOH) of the p53 gene in cirrhotic and neoplastic foci in the livers of patients with HCC. To assess the relationship between the LOH status of the p53 gene in the liver cirrhosis stage and that in HCC, we analyzed the samples microdissected from paraffin-embedded tissues using the polymerase-chain-reaction-based assay. We studied 18 patients with HCC. Fourteen of the 18 cases showed constitutional heterozygosity for the microsatellite markers. In 8 (57%) of the 14 informative cases, LOH was detected in primary HCCs. Among these 8 doubly informative (informative and LOH positive in primary HCC) cases, 5 cases (63%) showed LOH in liver cirrhosis lesions. The pattern of p53 allelic loss in the cirrhotic foci was identical with that in the corresponding tumor. The remaining 6 cases without LOH of the p53 gene in HCC showed on p53 loss in any cirrhotic foci. LOH of the p53 gene may occur before the development of HCC. PMID- 9216856 TI - Correlation between the lymphocytic infiltration of tumors and the proliferative activity of cancer cells from surgically treated esophageal carcinoma. AB - To evaluate the correlation among the biological behavior of cancer cells, the local immune response of the host, and survival of patients with esophageal carcinoma, we investigated the proliferative activity of cancer cells by immunostaining with the Ki-67 monoclonal antibody in 95 cases of surgically resected esophageal squamous cell carcinoma and compared them with the extent of lymphocytic infiltration of the tumor (LI) in the same specimens. The mean Ki-67 score of 43 tumors with low-grade LI was 46.7% and it was not different from that of 52 tumors with high-grade LI (49.4%, p = 0.441). A significant correlation between the Ki-67 score and the clinicopathological characteristics of tumors (depth of tumor invasion, lymph node metastasis, and stage) was detected. However, in the same stage, the survival of patients with a low Ki-67 score was not different from that of patients with a high Ki-67 score. On the other hand, even no significant correlation between the extent of LI and the clinicopathological characteristics of tumors was found; the 5-year survival rate of 17 patients with high-grade LI (13.9%) was significantly better than that of 15 patients with low-grade LI (0%, p = 0.07) in stage III. These facts indicate that proliferative activity might correlate with the tumor progression, however, when the survival is compared in the same stage, the local immune response such as the extent of lymphocytic infiltration of the tumor might be the better prognostic factor than proliferative activity in patients with esophageal cancer. PMID- 9216857 TI - Value of quantitative DNA analysis in endocrine tumors of the pancreas. AB - BACKGROUND: The diagnosis of malignancy can be difficult in endocrine tumors of the pancreas. Moreover prognostically relevant factors are not available. The aim of this study was to evaluate retrospectively whether the DNA distribution pattern can differentiate between benign and malignant pancreatic endocrine tumors and secondly whether the DNA content of tumor cells gives prognostic information. METHOD: Image cytometry of paraffin-embedded tumor material of 42 pancreatic endocrine tumors. RESULTS: In 27 benign endocrine pancreatic tumors (25 insulinomas, 2 benign nonfunctioning endocrine tumors) we could differentiate between 6 diploid, 15 hypotriploid and 6 triploid DNA histograms. In 15 malignant endocrine tumors of the pancreas we could differentiate between 1 diploid, 1 hypotriploid, 9 triploid and 4 hypertriploid tumors. All 4 patients with a hypertriploid tumor died as a consequence of their disease in contrast to only 1 patient with a diploid, hypotriploid or triploid tumor even in case of a nonradical resection. CONCLUSION: With the help of quantitative DNA measurement a differentiation between malignant and benign endocrine pancreatic tumors is not possible even if the risk of malignancy increases with the DNA content. The DNA content of malignant endocrine pancreatic tumors has an influence on the long term survival. Hypertriploid tumors have a statistically significantly worse prognosis than diploid, hypotriploid or triploid tumors. PMID- 9216858 TI - Angiogenesis and expression of platelet-derived endothelial cell growth factor in oral squamous cell carcinoma. AB - It has been demonstrated that angiogenesis is required in the process of tumor progression and metastasis. Microvessel density (MVD) estimates tumor angiogenesis and is an independent indicator for predicting tumor metastasis in a variety of carcinomas. Platelet-derived endothelial cell growth factor (PD-ECGF) is known to be an angiogenic factor in vitro and in vivo. Of 55 patients with oral squamous cell carcinoma (OSCC), regional metastasis was absent in 35 and present in 20. Cases with lymph node metastasis showed significantly higher MVD (mean 61.0 +/- 28.8) than those without metastasis (mean 29.3 +/- 15.1; p < 0.001). A total of 37 cases (67.3%) were PD-ECGF-positive with a high MVD (mean 47.8 +/- 27.9) and 18 (32.7%) showed a negative PD-ECGF expression with a low MVD (mean 26.6 +/- 13.2). PD-ECGF expression was significantly correlated with the increment of MVD (p < 0.01). We suggest that MVD can be used as an independent prognostic indicator for predicting metastasis and that PD-ECGF activity plays an important role in the neovascularization of OSCC. PMID- 9216859 TI - Induction of acquired immunity in rats that have eliminated intracranial gliosarcoma cells by the expression of herpes simplex virus-thymidine kinase gene and ganciclovir administration. AB - We examined a possible antitumor response against 9L rat gliosarcoma cells induced by the expression of the herpes simplex virus-thymidine kinase (HSV-TK) gene and the ganciclovir (GCV) system. Based on the amount of the major histocompatibility complex (MHC) class I antigens expressed on 9L cells transduced by the HSV-TK gene (9L/HSV-TK) we selected two clones (clones H and L), which represent high and low expressors of class I antigens, respectively. By means of serial magnetic resonance imaging we followed the change of tumor volumes of each clone in syngeneic rats, and found that the intracranial tumor growth was inversely correlated with the expression of MHC class I antigens, although in vitro growths of the clones remained unchanged. Moreover, histological examination revealed significant lymphocyte infiltration in the 9L/HSV-TK tumor of high MHC expression but not in the wild-type tumor. The therapeutic effect of GCV on them was not different, but we observed a prolonged survival of the rats which had eliminated 9L/HSV-TK clone L tumors by the treatment of GCV and were rechallenged with the same cells compared with the survival of naive rats inoculated with clone L cells. These data collectively suggest that the immune response operates even in the brain previously described as an immunologically privileged site. PMID- 9216860 TI - Comparison of spontaneous and induced mutation rates in an immortalized human bronchial epithelial cell line and its tumorigenic derivative. AB - To determine the relationship between neoplastic transformation and increased genetic instability, spontaneous and induced mutation rates were compared in a nontumorigenic, immortalized human bronchial epithelial cell line (NL20) and a tumorigenic cell line (NL20T) spontaneously derived from the NL20 line. Using the hypoxanthine phosphoribosyltransferase (HPRT) locus as a marker for determining mutation rate, fluctuation analysis was utilized to evaluate the spontaneous mutation rate. Induced mutation rates were determined for each cell line after N methyl-N'-nitro-N-nitrosoguanidine exposure. Both the spontaneous and induced mutation rates were noted to be significantly higher in the nontumorigenic NL20 cell line. These findings suggest that increasing genetic instability, as measured by spontaneous or induced mutation rate in the HPRT locus, does not correlate with tumorigenicity in these cells. PMID- 9216861 TI - Overexpression of metallothionein correlates with chemoresistance to cisplatin and prognosis in esophageal cancer. AB - The aim of this study was to determine the correlation of metallothionein (MT) expression with resistance to cisplatin and to identify prognostic factors in esophageal cancer. Immunostaining for MT was performed on the specimens of squamous cell carcinoma of the esophagus resected from 68 patients with curative intent. The expression of MT was evaluated in terms of clinicopathologic variables, effect of cisplatin, and the patients' survival. Overexpression of MT in the tumor was found in 70.6% of the patients. Stage III and IV tumors were more common in MT-positive tumors (p = 0.0282) but there was no difference in other clinicopathologic variables between MT-positive and MT-negative groups. Stage, cisplatin therapy, and tumor length were the independent prognostic indicators by multivariate analysis. Among 43 patients treated with cisplatin, the 5-year survival rate was 56% for MT-negative and 26% for MT-positive patients (p = 0.0277). In the MT-negative patients, multivariate analysis revealed that curability, stage, cisplatin therapy, and tumor length were the independent prognostic factors. These findings suggest that MT expression in squamous cell carcinoma of the esophagus is a major determinant of the chemoresistance to cisplatin and may be a predictor of poor prognosis. PMID- 9216862 TI - Undifferentiated nasopharyngeal carcinoma: pattern of failure--experience at the Johannesburg Hospital (1989-1994). AB - The records of 18 African patients with recurrent undifferentiated nasopharyngeal carcinoma (UDNPC) were reviewed. The skeletal system was the most common site of distant metastases. The pattern of skeletal involvement conformed to the general pattern, with the spine and pelvis being the most common sites. We conclude that metastatic UDNPC seen in Southern Africa resembles its North African and Southeast Asian counterparts. PMID- 9216863 TI - Molecular models for the stereochemical structures of fumonisin B1 and B2. AB - Assessment of the structural configuration of fumonisin B1 (FB1) and B2 (FB2) would allow better understanding of the behavior of these molecules during isolation or other handling procedures, and their interaction with binding sites or antibodies. Molecular models of the absolute configurations of FB1 and FB2 were calculated using a CAChe WorkSystemtrade mark. The electrostatic potential energy surfaces of the minimum potential energy conformations for FB1 and FB2 also were obtained. The models reveal that the backbone and acid side chains for FB1 and FB2 form a spherical globular model. The folded region forms a cage-like feature. It is this feature that suggests that these molecules may be potential chelators. The electrostatic potential surfaces show that most of the exposed surfaces of these molecules are hydrophobic in nature, and that there is a distinct difference between the electrostatic potential surfaces of the FB1 models resulting from the stereochemistry proposed by Shier et al. (1995) and Boyle and Kishi (1995a). The electrostatic surfaces clearly show a different orientation of the hydrophobic tail region of the backbone for FB2 than in the models for FB1. PMID- 9216864 TI - The effects of motorway runoff on freshwater ecosystems: 3. Toxicant confirmation. AB - Previous studies have demonstrated that small streams receiving road runoff have reduced water and sediment quality. These changes in quality are associated with alterations in the structure and functioning of stream communities. Laboratory studies have indicated that the community changes are due to sediment-associated contaminants, and toxicant identification evaluations have shown that the major toxicants are contained probably in a fraction of sediment extract that contains polycyclic aromatic hydrocarbons (PAHs). The aim of the present study was to determine whether PAHs were indeed the major toxicants in sediment extracts. Toxicity tests were performed with PAH mixtures, the toxic fraction of an extract of runoff-contaminated sediment, and a whole sediment extract. These indicated that three PAHs accounted for the toxicity of a sediment extract: pyrene, fluoranthene, and phenanthrene. The possibility of spatial or temporal variation in major toxicants was also investigated and tests on a number of sediment extracts obtained from a number of sites at different times demonstrated that the three PAHs accounted for 30.8 to 120% of an extract's toxicity. When the PAHs were considered individually, pyrene was shown to account for most of the toxicity (44.9%), followed by fluoranthene (16%) and phenanthrene (3.5%). PMID- 9216865 TI - The urgent need for environmental sanitation and a safe drinking water supply in Mbandjock, Cameroon. AB - Studies were conducted to assess the physical, chemical, and bacteriological qualities of drinking water in Mbandjock, Cameroon. Study results indicated that the vast majority of drinking water sources possessed acceptable physical and chemical qualities, according to the World Health Organization standards. However, microbiological analyses revealed that only the waters treated by the Cameroon National Water Company (SNEC) and the Sugar Processing Company (SOSUCAM) were acceptable for human consumption. All spring and well waters presented evidences of fecal contamination from human and/or animal origin. Water from these sources should, therefore, be treated before use for drinking. Since the majority of the population gets its water from wells and springs, there is an urgent need to develop a health education program, within the framework of primary health care, with respect to environmental sanitation and safe drinking water supply in this community. PMID- 9216866 TI - Diffusive sampling and biological monitoring of 2-bromopropane. AB - The possibilities to apply personal ambient air monitoring by diffusive sampling and biological exposure monitoring by urinalysis for 2-bromopropane or its metabolites were explored. The abilities of carbon cloth to adsorb 2-bromopropane was examined by experimental vapor exposure followed by solvent extraction and FID-GC. Urine from factory workers and rats exposed to 2-bromopropane were analyzed for 2-bromopropane, acetone and isopropyl alcohol by FID-GC, and for bromide ion by ECD-GC after chemical methylation. Carbon cloth adsorbed 2 bromopropane in a manner linearly related to exposures up to 1500 mg/m3 and to 8 h. The adsorption could quantitatively detect a 15 min peak exposure at 3,000 mg/m3. In rat experiments, analyses of urine samples collected over a 4-h period after termination of a 4-h exposure to 2-bromopropane at 500, 1,000 or 1,500 mg/m3 showed that acetone and bromide ion were excreted dose-dependently. Essentially, no 2-bromopropane or isopropyl alcohol was detected. When the analytical methods were applied to urine samples from 5 male workers exposed to 2 bromopropane at a low level (3 mg/m3 as a geometric mean), acetone and bromide ion levels were within respective normal ranges in four cases, but were higher than the upper limits of the normal ranges in the fifth case of a foreman who probably had the highest exposure. Thus, diffusive sampling is applicable to monitor exposure to 2-bromopropane. Urinalysis for acetone and bromide ion in combination appears to be a promising selective tool for biological monitoring of occupational exposure to 2-bromopropane. PMID- 9216867 TI - Recovery following pulsed exposure to organophosphorus and carbamate insecticides in the midge, Chironomus riparius. AB - The importance of recovery following pulsed and continuous exposure was determined by measuring the acute toxicity of two organophosphorus (parathion and malathion) and four carbamate (aldicarb, carbaryl, carbofuran and propoxur) insecticides. Two 1-h pulses caused significantly fewer symptoms of intoxication than 2 h of continuous exposure if at least 2 to 6 h in clean water were provided between doses for the four carbamates. Two 1-h pulses were equally toxic as a single 2-h continuous exposure for the two organophosphorus insecticides. Acetylcholinesterase activity in midges given two 1-h pulses of carbaryl separated by 24 h in clean water showed reactivation to control levels between the two exposures. These results contribute to the belief that episodic exposure to insecticides is less toxic if recovery in clean water is provided. PMID- 9216868 TI - Effect of soil moisture on pesticide toxicity to an enchytraeid worm, Enchytraeus sp. AB - The aim of the study was to find out whether soil moisture affects toxicity of organic pesticides to an enchytraeid worm. Laboratory experiments were carried out with dimethoate and benomyl, using a small Enchytraeus sp. as the test species. Substrate was natural agricultural field soil cultivated without pesticides for several years. Experimental design consisted of three soil moistures (40, 55, and 70% of water holding capacity) and five pesticide concentrations, plus controls. Measured parameters were survival, size of the parent worms and number and size of juveniles produced. Dimethoate was relatively non-toxic to this species. Dimethoate did not decrease survival, but sublethal effects on adult size and number of juveniles were observed. Adverse conditions in dry soil masked these effects; dimethoate appeared to be less toxic in dry soil than in moist soil. Benomyl caused significant mortality and the effects were very abrupt. Toxicity of benomyl decreased with increasing soil moisture content; in moist soil the worms survived at higher benomyl concentrations than in drier soils. PMID- 9216871 TI - Metallothionein-like protein: is It an efficient biomarker of metal contamination? A case study based on fish from the Tunisian coast. AB - The aim of this work was to assess the relative importance of natural fluctuations in metallothionein-like protein (MTLP) levels associated with the sexual status of fish versus fluctuations due to metal exposure. In order to see fluctuations due to metal exposure, comparisons were made on the same fish species Scorpaena porcus sampled in polluted and unpolluted sites. The hermaphrodite fish Serranus scriba and Scorpaena porcus, in which sexes are separate, were compared at the unpolluted site to see fluctuations caused by the sexual status. In both species, metals and the MTLP levels were determined in the gills and liver. In these organs, Cd, Cu, and Zn distributions were examined in different fractions: the insoluble fraction (IF) and the cytosol divided into thermolabile compounds (HDF) and the heat stable compounds including MTLP. MTLP levels were higher in the liver (3.09 mg/g in S. porcus, 1.59 mg/g in S. scriba) than in gills (0.13 mg/g in S. porcus, 0.40 mg/g in S. scriba). For Scorpaena porcus, metals and MTLP levels varied with sex, whereas in Serranus scriba, which is a hermaphrodite species, inherent variations were also observed. At the polluted site, MTLP, Cd, and Cu concentrations in the gills of S. porcus increased but the supplementary metals were not associated with the heat stable compounds including MTLP. At this site, hepatic MTLP bound more metals than at the unpolluted site, but its binding capacity was not sufficient to avoid the binding of metals to the insoluble and the heat denaturable fractions. In light of these results and in spite of its hermaphrodism, it is questionable whether to consider S. scriba as a good candidate for biomonitoring based on MTLP. S. porcus could be useful for this purpose only if the MTLP capacity in binding metals is not exceeded. The MTLP could be considered as a biomarker only if it is investigated in relatively unpolluted sites. PMID- 9216869 TI - Organophosphates in the zebra mussel Dreissena polymorpha: subacute exposure, body burdens, and organ concentrations. AB - Subacute exposures (10 d) of the freshwater mollusc Dreissena polymorpha to disulfoton (10 mg/L), thiometon (6 mg/L), and its activated oxygen analogue demeton-S-methyl (6 mg/L) corroborate earlier findings of organophosphate resistance and accumulation in the organism. Mortality occurred not before the ninth day of exposure. Mortality was induced at high ambient water concentrations and must be due to unknown specific organophosphate effects. Body burdens reached saturation levels within one week being around 40 mg/kg wet weight for thiometon and 60 mg/kg for disulfoton. Mussels dying during the tests showed lower tissue concentrations. Elimination of accumulated organophosphates was so low in the mussel, that an efficient metabolism of these compounds in the mussel was unlikely. Different organs of Dreissena previously acutely exposed (96 h) to the organophosphate thiometon (6, 12, 25, 50 mg/L) were analyzed for their thiometon content. Thiometon could be found in all organs, but were highest in the anterior part of the viscera (230 mg/kg), where it was accumulated either in the digestive gland and/or in the gonadal tissue. PMID- 9216872 TI - Spatial patterns in a bioindicator: heavy metal and selenium concentration in eggs of herring gulls (Larus argentatus) in the New York Bight. AB - Concentrations of selenium and five heavy metals (lead, cadmium, mercury, chromium, and manganese) in the eggs of herring gulls (Larus argentatus) were studied at six breeding colonies in the New York Bight to detect locational differences and to explore their use as a bioindicator of point source or nonpoint source pollution. The herring gull is widespread in North America, Europe, and Asia, and has urban-adapted counterparts in the southern hemisphere as well. We anticipated that the chromium contamination at Jersey City and high levels of manganese in industrial releases to the Passaic River would be reflected in the nearest colony (Shooter's Island), and that lead contamination from bridge remediation would be apparent in the Jamaica Bay colonies. There were significant locational differences in all metal levels, although the patterns were not the same for all metals. Shooter's Island in Newark Bay ranked first or second for five of the elements, but inexplicably had the lowest mercury level. Cadmium levels were highest at Canarsie Pol in Jamaica Bay, but mercury levels were highest at the relatively isolated Lavallette colony in northern Barnegat Bay. Chromium and manganese levels were indeed highest at Shooter's Island, but the lead levels in Jamaica Bay were only intermediate. We predicted that the essential trace elements, manganese, chromium, and selenium, which are known to be present at relatively high concentrations in various animal species, would have relatively low coefficients of variation, reflecting homeostatic mechanisms. This was confirmed. In conclusion, herring gull egg contents can be used to monitor metal concentrations at nearby colonies to indicate areas of concern for particular metals. They may confirm suspected associations or identify hitherto unsuspected problems. PMID- 9216874 TI - Environmental exposure and distribution of lead in four species of raptors in Southeastern Spain. AB - The purpose of this study was to monitor exposure to lead in four species of raptors in Southeastern Spain (Murcia Region). Samples of liver, kidney, brain, blood, and bone from two species of diurnal raptors (European kestrel (Falco tinnunculus) and European buzzard (Buteo buteo)) and two species of nocturnal raptors (Eagle owl (Bubo bubo) and Little owl (Athene noctua)) were obtained during 1994. Relationships were found between size and age of the birds, the nearness to areas of human activity and lead concentrations in tissues. The lead distribution pattern reveals that the bone is the principle organ for accumulation (0.62-43 mg/Kg, dry weight), followed by the kidney (0.03-0.66 mg/Kg, wet weight), and liver (0. 017-0.05 mg/Kg, w.w.), and to lesser extent, the brain (0.013-0.223 mg/Kg, w.w.). This distribution pattern indicates that raptors in Southeastern Spain were exposed to environmental low lead levels continuously over an extended period of time. Correlations between lead in bone and lead in soft tissues were higher in European buzzards (r = 0.87-0.95) and Eagle owl (r = 0.71-0.86) than those found in European kestrels (r = 0.53-0.58) and Little owls (r << 0). However, correlations between lead concentrations in soft tissues and in blood were high (r = 0.85-0.99). PMID- 9216875 TI - Mixed function oxidases in an Australian marsupial, the brushtail possum (Trichosurus vulpecula). AB - Investigation of the mixed function oxidase system of the brushtail possum was undertaken to provide fundamental information about this detoxication enzyme system in a marsupial. Brushtail possum hepatic cytochrome P450, cytochrome b5 and NADPH-cytochrome c reductase levels, 7-ethoxyresorufin O-deethylase and (EROD) 7-ethoxycoumarin O-deethylase (ECOD) activities were in the range of values reported for eutherian mammals. Hepatic cyrochrome P450 content was significantly greater (p < 0.01) in brushtail possums from a non-urban population in comparison to an urban population, as was ECOD activity (p < 0.0001). EROD activity was significantly greater in female brushtail possums in comparison to males (p < 0.01). The factors potentially influencing the population- and sex specific expression of cytochrome P450 isoenzymes in the brushtail possum are discussed and include exogenous dietary xenobiotics and endogenous hormonal alterations influenced by reproductive status. PMID- 9216876 TI - Effect of the day of administration on the developmental toxicity of tributyltin chloride in rats. AB - The objective of this study was to determine the susceptible day for the teratogenicity of tributyltin chloride (TBTCI) by a single administration on one of the days during organogenesis. Pregnant rats were given a single dose of TBTCI by gastric intubation at 100 mg/kg on either day 7, day 8, or day 9 and at 200 mg/kg on either day 7, day 8, day 9, day 10, day 11, day 12, day 13, day 14, or day 15 of pregnancy. The maternal body weight gain in the period immediately following administration in all TBTCI-treated groups was significantly decreased. A significant increase in the incidence of postimplantation loss was found after administration of TBTCI on day 7, day 8, and day 9 at 100 and 200 mg/kg and on day 10 and day 11 at 200 mg/kg. A significantly increased incidence of fetuses with external malformations was detected when TBTCI was given on day 8 at 100 and 200 mg/kg and on day 11, day 12, day 13, and day 14 at 200 mg/kg, and the most pronounced effect occurred after administration on day 13 of pregnancy. Cleft palate was observed exclusively after administration during late organogenesis. It could be concluded that the manifestation and susceptibility of the developmental toxicity of TBTCI vary with the developmental stages at the time of administration and that TBTCI has the biphasic sensitivity to teratogenicity on day 8 and days 11-14 of pregnancy. PMID- 9216877 TI - Variation of PCB congener levels during lactation period and relationship to their molecular structure. AB - The levels of 22 polychlorinated biphenyl (PCB) congeners, including 13 coplanars (non-, mono-, and di-ortho-substituted chlorine) and 9 noncoplanar chirals (tri- and tetra-ortho-substituted chlorine), were measured in mothers milk from different mothers throughout their lactation periods. Important variations were found in the total PCB levels and PCB congener levels. The maximum PCB variations were found during the first weeks of lactation. Of the 22 PCBs analyzed, the most abundant, and those which had the highest variations were PCB-101, 118, 138, 151, 170, and 180, while the least abundant were the PCB-77, 126, 169, and 167, which also showed the lowest variation. Among the nonortho-substituted PCB congeners, PCB-77 was the predominant PCB in three of the four lactation periods studied, while PCB-126 was dominant in the last case. A relationship has been found between the levels and variations of total PCBs in the milk supplied by the different mothers and their dietary habits, weight changes and illness suffered during their pregnancy and lactation. The 22 PCB congeners investigated have been grouped in five categories according to their concentration variations in milk throughout the four different lactation periods. The PCB congeners included in each of the five groups have similar molecular structures. It has been found that coplanarity, number of chlorines and their molecular distribution, and neighbor H atoms were determinant factors in the process of PCB mobilization from the mothers' fats (where it had been able to accumulate throughout her life in the case of her first child) to the breast milk she delivered. PMID- 9216878 TI - Blood serum levels of PCBs and PCDFs in Yucheng women 14 years after exposure to a toxic rice oil. AB - In 1979, a mass poisoning of more than 2000 people occurred in central Taiwan due to consumption of rice-bran oil contaminated with PCBs and their heat-degraded byproducts. The incident was later referred to as Yucheng (oil disease). Serum samples from 56 women with the 1979 exposure were collected in February 1992 and analyzed for their contaminant content using sample enrichment and isotope dilution mass spectrometry. In most of the samples, levels of PCDFs and PCBs were detectable, and the median values of 2,3,4,7,8-PCDFs and 1,2,3,4,7,8-PCDFs were 1,030 and 2,220 ng/kg serum lipid, respectively. The median level of the total PCBs on a whole weight basis was 8,730 ng/kg. The PCB/PCDF concentrations in Yucheng women 14 years after the toxic exposure were still one to two orders of magnitude higher than controls. Concentrations of PCB levels in 1992 were positively correlated with the 1980-1981 measured PCB levels in these women and both PCBs and PCDFs were negatively correlated with the total duration when these women breast fed their children between 1979 and 1992. It is concluded that serum levels of congener-specific PCBs/PCDFs in exposed women are good indicators of previous exposure and may provide important information for more reliable estimation of dose-response relationship. PMID- 9216879 TI - Isolation of Bacillus megaterium mutants that produce high levels of heterologous protein, and their use to construct a highly mosquitocidal strain. AB - A xylose-regulated plasmid expression system for producing high levels of recombinant proteins in Bacillus megaterium has recently been described [Appl Microbiol Biotechnol 35:594, 1991]. Using an antibiotic resistance protein as the expressed protein, we have been able to select mutant plasmids that produce increased levels of heterologous protein. The mutant plasmids show increased segregational stability and have lost the ability to be transformed into Escherichia coli. The same selection protocol has been used to isolate a mutant strain producing high levels of the Bacillus sphaericus mosquitocidal binary toxin. This strain shows toxicity to Culex quinquefasciatus larvae that is comparable toB. sphaericus 2362 and higher than a B. megaterium strain with the original expression plasmid. This approach may be generally useful for high-level regulated protein expression in B. megaterium. PMID- 9216880 TI - Aerobic gram-positive and gram-negative bacteria exhibit differential sensitivity to and transformation of 2,4,6-trinitrotoluene (TNT). AB - A systematic evaluation of the ability of different bacterial genera to transform 2,4,6-trinitrotoluene (TNT), and grow in its presence, was conducted. Aerobic Gram-negative organisms degraded TNT and evidenced net consumption of reduced metabolites when cultured in molasses medium. Some Gram-negative isolates transformed all the initial TNT to undetectable metabolites, with no adsorption of TNT or metabolites to cells. Growth and TNT transformation capacity of Gram positive bacteria both exhibited 50% reductions in the presence of approximately 10 microg TNT ml-1. Most non-sporeforming Gram-positive organisms incubated in molasses media amended with 80 microg TNT ml-1 became unculturable, whereas all strains tested remained culturable when incubated in mineral media amended with 98 microg TNT ml-1, indicating that TNT sensitivity is linked to metabolic activity. These results indicate that the microbial ecology of soil may be severely impacted by TNT contamination. PMID- 9216881 TI - The (F1F0) ATP synthase of Buchnera aphidicola (endosymbiont of aphids): genetic analysis of the putative ATP operon. AB - Buchnera aphidicola is an intracellular, non-cultivable prokaryotic symbiont of the aphid Schizaphis graminum. A 6.8-kilobase fragment from B. aphidicola was cloned and sequenced and was found to contain genes encoding for proteins of the ATP synthase. The order of the genes, atpBEFHAGDC, is identical to that found inEscherichia coli and many other prokaryotes. This genetic organization is different from that observed in organelles such as mitochondria and chloroplasts, in which the genes are partitioned between the organellar and nuclear genomes. One difference between B. aphidicola and E. coli was the absence of atpI, a gene of unknown function, which in E. coli precedes atpB. As is the case of many other prokaryotes, atpBEFHAGDC appears to constitute a single transcription unit. The detection in B. aphidicola of the genes encoding the ATP synthase as well as past observations indicating that this organism is capable of respiration are consistent with the utilization byB. aphidicola of a proton gradient for the generation of ATP. PMID- 9216882 TI - The effect of nisin and monensin on ruminal fermentations In vitro. AB - When mixed ruminal bacteria and alfalfa were incubated in vitro, monensin and nisin both inhibited methane production so long as the concentrations were greater than 1 microM. Monensin- and nisin-dependent methane depressions caused a decrease in the acetate to propionate ratio (4.5 to 3.0). Total volatile fatty acid production was decreased by both monensin and nisin addition at concentrations greater than 2 microM. Starch-digesting ruminal bacteria were initially inhibited by monensin and nisin, but this effect disappeared after two to four transfers. Nisin always inhibited cellulolytic bacteria, but the nisin dependent inhibition of cellulose digestion was no greater than the inhibition caused by monensin. Monensin and nisin also inhibited amino acid degradation, and nisin was more effective than monensin in controlling the growth of Clostridium aminophilum, an obligate amino acid-fermenting ruminal bacterium that can tolerate low concentrations of monensin. Because nisin was as potent as monensin, bacteriocins such as nisin may have potential as feed additives. PMID- 9216883 TI - Detection and molecular analysis of plant- and insect-associated bacteria harboring aconitate isomerase involved in biosynthesis of trans-aconitic acid as antifeedant in brown planthoppers. AB - The activity of aconitate isomerase, which is involved in the biosynthesis of trans-aconitic acid as antifeedant in brown planthoppers, was detected in Pseudomonas fluorescens LRB3W1 and Pseudomonas putida MAFF301685 but not in Pseudomonas putida MAFF301684. The enzyme activity was induced in the presence of trans-aconitate, and therefore bacteria showing the enzyme activity were easily detected by their ability to grow on the minimal medium containing trans aconitate as the sole carbon source (ACO agar medium). Experiments on growth of plant- or insect-associated bacteria on ACO agar medium showed that most of the Gram-negative bacteria displayed the aconitate isomerase activity unlike most of the Gram-positive bacteria isolated mainly from insects. Mini-Tn5 transposon derivatives of P. fluorescens LRB3W1 lacking completely or partially their ability to grow on ACO agar medium were obtained. Southern blot analysis with a mini-Tn5 DNA probe definitely showed that the genes responsible for the biosynthesis of aconitate isomerase present on chromosomal DNA. Thus, it was suggested that genes for aconitate isomerase biosynthesis are commonly present in Gram-negative plant- or insect-associated bacteria, and also the DNA fragments including the genes were detected in P. fluorescens LRB3W1. PMID- 9216884 TI - The role of fluoroquinolones in the promotion of alginate synthesis and antibiotic resistance in Pseudomonas aeruginosa. AB - Treatment of nonmucoid Pseudomonas aeruginosa with gyrase inhibitors such as ciprofloxacin, norfloxacin, and ofloxacin, which target the A subunit of topoisomerase II, resulted in 100% conversion to the mucoid phenotype. However, antibiotics that partially inhibited growth and macromolecular synthesis (DNA, RNA, protein, or peptidoglycan) of nonmucoid isolates in a gluconate-limited chemostat culture system did not promote conversion to mucoid subpopulations. An increase in resistance was observed in populations that expressed the mucoid phenotype. Both mucoid conversion and antibiotic resistance were completely reversible when ciprofloxacin pressure was withdrawn, but only partially reversible by the removal of norfloxacin and ofloxacin. Thus, these experiments indicate that in the presence of some fluoroquinolones, a conditional response resulting in mucoid conversion and antibiotic resistance may occur. PMID- 9216885 TI - Isolation and characterization of Vibrio carchariae, a causative agent of gastroenteritis in the groupers, Epinephelus coioides. AB - An outbreak of serious mortality among the cultured groupers Epinephelus coioides, characterized by a swollen intestine containing yellow fluid, occurred in the summer of 1993 in Taiwan. A motile strain EmI82KL was isolated from the intestinal yellow fluid of the moribund groupers with tryptic soy agar supplemented with 2% NaCl and/or thiosulfate citrate bile salt sucrose agar. This strain was characterized and identified as Vibrio carchariae and was susceptible to chloramphenicol, doxycycline-HCl, nalidixic acid, oxolinic acid, oxytetracycline, and sulfonamide while resistant to ampicillin and penicillin G. In addition, the strain was neither auto-agglutinating nor hemagglutinating, but it was hemolytic against erythrocytes from sheep, rabbit, tilapia, and grouper. The bacteria could be reisolated from kidney, liver, and the transparent yellow fluid of swollen intestine of moribund groupers after bacterial challenge and re identified as the same species. The LD50 value was 2.53 x 10(7) colony forming units/g grouper body weight. PMID- 9216886 TI - Heat-shock response in Methanosarcina mazei S-6. AB - The dnaK locus of Methanosarcina mazei S-6, a mesophilic organism of the phylogenetic domain Archaea, contains the heat-shock genes 5'-grpE-dnaK-dnaJ-3'. Parameters known to affect the response of these genes in organisms of the other two domains, Bacteria and Eucarya, were tested to determine their effects on the archaeal homologs. The mRNA from the three genes increased after heat shock more in lamina than in single cells (these S-6 morphologic stages can be grown in the same substrate). Single cells in early stationary phase showed the highest levels of dnaK mRNA after heat shock, as compared with cells in exponential, or in late stationary, phase. The dnaK mRNA always had the size of a monocistronic transcript. dnaK was also found in the thermophileMethanosarcina thermophila TM 1, and its response to heat shock showed distinctive characteristics. However, dnaK was not revealed in other archaea: three hyperthermophiles (Methanothermus fervidus,Methanococcus jannaschii, and Sulfolobus sp.), and one mesophilic methanogen (Methanospirillum hungateii). PMID- 9216887 TI - Buchnera aphidicola (endosymbiont of aphids) contains nuoC(D) genes that encode subunits of NADH dehydrogenase. AB - A two-kilobase DNA fragment from Buchnera aphidicola, the endosymbiont of aphids, was cloned and sequenced. One open reading frame was detected, coding for a putative protein of 600 amino acids. The N-terminal portion of this protein corresponded to NuoC, while the C-terminal portion corresponded to NuoD. These proteins are constituents of the membrane-associated NADH dehydrogenase. Our results suggest that these two proteins are fused in Buchnera aphidicola, a result consistent with their previously postulated spatial association. PMID- 9216888 TI - The effect of ethanol and oxygen on the growth of Zymomonas mobilis and the levels of hopanoids and other membrane lipids. AB - Zymomonas mobilis (ATCC 29191) was grown either aerobically or anaerobically in the presence of 2% (wt/vol) glucose and 0, 3, or 6% (vol/vol) ethanol. The rates of growth and the composition of hopanoids, cellular fatty acids, and other lipids in the bacterial membranes were quantitatively analyzed. The bacterium grew in the presence of 3% and 6% ethanol and was more ethanol tolerant when grown anaerobically. In the absence of ethanol, hopanoids comprised about 30% (by mass) of the total cellular lipids. Addition of ethanol to the media caused complex changes in the levels of hopanoids and other lipids. However, there was not a significant increase in any of the hopanoid lipid classes as ethanol concentration was increased. As previously reported, vaccenic acid was the most abundant fatty acid in the lipids of Z. mobilis, and its high constitutive levels were unaffected by the variations in ethanol and oxygen concentrations. A cyclopropane fatty acid accounted for 2.6-6.4 wt % of the total fatty acids in all treatments. PMID- 9216936 TI - Invited editorial on "Acute and chronic effects of exercise on leptin levels in humans". PMID- 9216937 TI - Acute and chronic effects of exercise on leptin levels in humans. AB - The acute (single bout of exercise) and chronic (exercise training) effects of exercise on plasma leptin were investigated in 97 sedentary adult men (n = 51) and women (n = 46) participating in the HERITAGE Family Study. Exercise training consisted of a standardized 20-wk endurance training program performed in the laboratory on a computer-controlled cycle ergometer. Maximal oxygen uptake, body composition assessed by hydrostatic weighing, and fasting insulin level were also measured before and after training. Pre- and posttraining blood samples were obtained before and after completion of a maximal exercise test on the cycle ergometer. Exercise training resulted in significant changes in maximal oxygen uptake (increase in both genders) and body composition (reduction of fat mass in men and increase in fat-free mass in women). There were considerable interindividual differences in the leptin response to acute and chronic effects of exercise, some individuals showing either increase or reduction in leptin, others showing almost no change. On average, leptin levels were not acutely affected by exercise. After endurance training was completed, leptin levels decreased significantly in men (from 4.6 to 3.9 ng/ml; P = 0.004) but not in women. However, after the training-induced changes in body fat mass were accounted for, the effects of exercise training were no longer significant. Most of the variation observed in leptin levels after acute exercise or endurance training appears to be within the confidence intervals of the leptin assay. We conclude that there are no meaningful acute or chronic effects of exercise, independent of the amount of body fat, on leptin levels in humans. PMID- 9216938 TI - Restricted postexercise pulmonary diffusion capacity and central blood volume depletion. AB - Pulmonary diffusion capacity for carbon monoxide (DLCO), regional electrical impedance (Z0), and the distribution of technetium-99m-labeled erythrocytes together with concentration of plasma atrial natriuretic peptide (ANP) were determined before and after a 6-min "all-out" row in nine oarsmen and in six control subjects. Two and one-half hours after exercise in the upright seated position, DLCO was reduced by 6 (-2 to 21; median and range) %, the thoracic-to thigh electrical impedance ratio (Z0 thorax/Z0 thigh) rose by 14 (-1 to 29) %, paralleled by a 7 (-3 to 11) % decrease and a 3 (-5 to 12) % increase in the thoracic and thigh blood volume, respectively. These responses were associated with a decrease in the plasma ANP concentration from 15 (13-31) to 12 (9-27) pmol/l (P < 0.05). Similarly, in the supine position, Z0 thorax/Z0 thigh increased by 10 (-5 to 28) % when DLCO was reduced 12 (6-26) % (P < 0.05), whereas DLCO remained stable in the control group. The increase in Z0 thorax/Z0 thigh and the corresponding redistribution of the blood volume in both body positions show that approximately one-half of the postexercise reduction of DLCO is explained by a decrease in the pulmonary blood volume. The role of a reduced postexercise central blood volume is underscored by the lower plasma ANP, which aids in upregulating the blood volume after exercise in athletes. PMID- 9216939 TI - Endotoxin priming of thromboxane-related vasoconstrictor responses in perfused rabbit lungs. AB - In prior studies of perfused lungs, endotoxin priming markedly enhanced thromboxane (Tx) generation and Tx-mediated vasoconstriction in response to secondarily applied bacterial exotoxins. The present study addressed this aspect in more detail by employing precursor and intermediates of prostanoid synthesis and performing functional testing of vasoreactivity and measurement of product formation. Rabbit lungs were buffer perfused in the absence or presence of 10 ng/ml endotoxin. Repetitive intravascular bolus applications of free arachidonic acid provoked constant pulmonary arterial pressor responses and constant release reactions of TxA2 and prostaglandin (PG) I2 in nonprimed lungs. Within 60-90 min of endotoxin recirculation, which provoked progressive liberation of tumor necrosis factor-alpha but did not effect any hemodynamic changes by itself, both pressor responses and prostanoid release markedly increased, and both events were fully blocked by cyclooxygenase (Cyclo) inhibition with acetylsalicylic acid (ASA). The unstable intermediate PGG2 provoked moderate pressor responses, again enhanced by preceding endotoxin priming and fully suppressed by ASA. Vasoconstriction also occurred in response to the direct Cyclo product PGH2, again amplified after endotoxin pretreatment, together with markedly enhanced liberation of TxA2 and PGI2. In the presence of ASA, the priming-related increase in pressor responses and the prostanoid formation were blocked, but baseline vasoconstrictor responses corresponding to those in nonprimed lungs were maintained. Pressor responses to the stable Tx analog U-46619 were not significantly increased by endotoxin pretreatment, but some generation of TxA2 and PGI2 was also noted under these conditions. We conclude that endotoxin priming exerts profound effects on the lung vascular prostanoid metabolism, increasing the readiness to react with Tx-mediated vasoconstrictor responses to various stimuli, suggesting that enhanced Cyclo activity is an important underlying event. PMID- 9216940 TI - Exaggerated pulmonary hypertension with monocrotaline in rats susceptible to chronic mountain sickness. AB - Hilltop (H) strain Sprague-Dawley rats are more susceptible to chronic mountain sickness than are the Madison (M) strain rats. It is unclear what role pulmonary vascular remodeling, polycythemia, and hypoxia-induced vasoconstriction play in mediating the more severe pulmonary hypertension that develops in the H rats during chronic hypoxia. It is also unclear whether the increased sensitivity of the H rats to chronic mountain sickness is specific for a hypoxia effect or, instead, reflects a general propensity toward the development of pulmonary hypertension. Monocrotaline (MCT) causes pulmonary vascular remodeling and pulmonary hypertension. We hypothesized that the difference in the pulmonary vascular response to chronic hypoxia between H and M rats reflects an increased sensitivity of the H rats to any pulmonary hypertensive stimuli. Consequently, we expected the two strains to also differ in their susceptibility to MCT-induced pulmonary hypertension. Pulmonary arterial pressures in conscious H and M rats were measured 3 wk after a single dose of MCT, exposure to a simulated high altitude of 18,000 ft (barometric pressure = 380 mmHg), and administration of a single dose of saline as a placebo. The H rats had significantly higher pulmonary arterial pressures and right ventricular weights after MCT and chronic hypoxia than did the M rats. The H rats also had more pulmonary vascular remodeling, i.e., greater wall thickness as a percentage of vessel diameter, after MCT and chronic hypoxia than did the M rats. The H rats had significantly lower arterial PO2 than did the M rats after MCT, but the degree of hypoxemia was mild [arterial PO2 of 72.5 +/- 0.8 (SE) Torr for H rats vs. 77.4 +/- 0.8 Torr for M rats after MCT]. The H rats had lower arterial PCO2 and larger minute ventilation values than did the M rats after MCT. These ventilatory differences suggest that MCT caused more severe pulmonary vascular damage in the H rats than in the M rats. These data support the hypothesis that the H rats have a general propensity to develop pulmonary hypertension and suggest that differences in pulmonary vascular remodeling account for the increased susceptibility of H rats, compared with M rats, to both MCT and chronic hypoxia-induced pulmonary hypertension. PMID- 9216941 TI - Oxidation of lactate and acetate in rat skeletal muscle: analysis by 13C-nuclear magnetic resonance spectroscopy. AB - The balance between carbohydrate and fatty acid utilization in skeletal muscle previously has been studied in vivo by using a variety of methods such as arteriovenous concentration differences and radioactive isotope tracer techniques. However, these methodologies provide only indirect estimates of substrate oxidation. We used 13C-nuclear magnetic resonance (NMR) spectroscopy and non-steady-state isotopomer analysis to directly quantify the relative oxidation of two competing exogenous substrates in rat skeletal muscles. We infused [1,2-13C]acetate and [3-13C]lactate intravenously in anesthetized rats during the final 30 min of 35 (n = 10) or 95 (n = 10) min of intense, unilateral, rhythmic hindlimb contractions. 13C-NMR spectroscopy and isotopomer analysis were performed on extracts of gastrocnemius and soleus muscles from both the contracting and contralateral resting hindlimbs. We found that 1) [13C]lactate and [13C]acetate were taken up and oxidized by both resting and contracting skeletal muscles; and 2) high-intensity muscle contractions altered the pattern of substrate utilization such that the relative oxidation of acetate decreased while that of lactate remained unchanged or increased. Based on these findings, we propose that 13C-NMR spectroscopy in combination with isotopomer analysis can be used to study the general dynamics of substrate competition between carbohydrates and fats in rat skeletal muscle. PMID- 9216942 TI - Neuromuscular factors contributing to in vivo eccentric moment generation. AB - Muscle series elasticity and its contribution to eccentric moment generation was examined in humans. While subjects [male, n = 30; age 26.3 +/- 4.8 (SD) yr; body mass 78.8 +/- 13.1 kg] performed an isometric contraction of the knee extensors at 60 degrees of knee flexion, a quick stretch was imposed with a 12 degrees step displacement at 100 degrees /s. The test was performed at 10 isometric activation levels ranging from 1.7 to 95.2% of maximal voluntary contraction (MVC). A strong linear relationship was observed between the peak imposed eccentric moment derived from quick stretch and the isometric activation level (y = 1.44x + 7.08; r = 0.99). This increase in the eccentric moment is consistent with an actomyosin-dependent elasticity located in series with the contractile element of muscle. By extrapolating the linear relationship to 100% MVC, the predicted maximum eccentric moment was found to be 151% MVC, consistent with in vitro data. A maximal voluntary, knee extensor strength test was also performed (5-95 degrees, 3 repetitions, +/-50, 100, 150, 200, and 250 degrees/s). The predicted maximum eccentric moment was 206% of the angle- and velocity-matched, maximal voluntary eccentric moments. This was attributed to a potent neural regulatory mechanism that limits the recruitment and/or discharge of motor units during maximal voluntary eccentric contractions. PMID- 9216944 TI - Almitrine and doxapram decrease fatigue and increase subsequent recovery in isolated rat diaphragm. AB - The effects of almitrine bimesylate and doxapram HCl on isometric force produced by in vitro rat diaphragm were studied during direct muscle activation at 37 degrees C. Doxapram and almitrine ameliorate respiratory failure clinically by indirectly increasing phrenic nerve activity. This study was carried out to investigate possible direct actions of these agents on the diaphragm before and after fatigue of the fibers. Two age groups of animals were chosen [6-14 wk (group 1) and 50-55 wk (group 2)] because it is known that increasing age decreases a muscle fiber's resistance to fatigue. Muscle strips were isolated from both group 1 and group 2 and directly stimulated (2-ms pulse duration, 5-15 V) to produce twitch tensions of 1.3 and 2.1 N/cm2, respectively. At low concentrations, doxapram ( O > L, whereas maximum velocity for oxidative enzymes (CS, cytochrome-c oxidase) was lowest in subjects with NIDDM. The ratio between glycolytic and oxidative enzyme activities within skeletal muscle correlated negatively with insulin sensitivity. The HK/CS ratio had the strongest correlation (r = -0.60, P < 0.01) with insulin sensitivity. In summary, an imbalance between glycolytic and oxidative enzyme capacities is present in NIDDM subjects and is more severe than in obese or lean glucose-tolerant subjects. The altered ratio between glycolytic and oxidative enzyme activities found in skeletal muscle of individuals with NIDDM suggests that a dysregulation between mitochondrial oxidative capacity and capacity for glycolysis is an important component of the expression of insulin resistance. PMID- 9216961 TI - Effects of vigorous exercise training and beta-agonist administration on bone response to hindlimb suspension. AB - The effectiveness of dobutamine (Dob) in preventing bone loss during 14 days of hindlimb suspension (Sus) was tested in exercise-trained (Ex; n = 25) and sedentary (Sed; n = 22) rats (age 155 days). One-half of each group was given Dob (2 mg . kg-1 . day-1) or saline (Sal). Histomorphometric measurements at midfemur revealed a 17% smaller cortical bone area (CBA) and a 32% lower periosteal mineral apposition rate (MAR) in suspended vs. nonsuspended Sed/Sal rats. Dob abolished this decline in CBA in Sed/Sus rats, probably via an attenuation of the decrease in periosteal MAR; similar but nonsignificant effects on cross-sectional moment of inertia were observed. Nonsuspended Ex rats had no change in bone CBA when CBA is indexed to body weight. Sus appeared to uncouple the relationship between soleus weight and CBA. Dob attenuated the 43% decline in soleus weight after Sus in Ex but not in Sed rats. In summary, vigorous Ex before Sus does not affect loss of bone mass due to unloading; Dob effectively maintains CBA in Sed rats subjected to suspension. PMID- 9216962 TI - Respiratory impedances and acinar gas transfer in a canine model for emphysema. AB - We examined how the changes in the acini caused by emphysema affected gas transfer out of the acinus (Taci) and lung and chest wall mechanical properties. Measurements were taken from five dogs before and 3 mo after induction of severe bilateral emphysema by exposure to papain aerosol (170-350 mg/dose) for 4 consecutive wk. With the dogs anesthetized, paralyzed, and mechanically ventilated at 0.2 Hz and 20 ml/kg, we measured Taci by the rate of washout of 133Xe from an area of the lung with occluded blood flow. Measurements were repeated at positive end-expiratory pressures (PEEP) of 10, 5, 15, 0, and 20 cmH2O. We also measured dynamic elastances and resistances of the lungs (EL and RL, respectively) and chest wall at the different PEEP and during sinusoidal forcing in the normal range of breathing frequency and tidal volume. After final measurements, tissue sections from five randomly selected areas of the lung each showed indications of emphysema. Taci during emphysema was similar to that in control dogs. EL decreased by approximately 50% during emphysema (P < 0.05) but did not change its dependence on frequency or tidal volume. RL did not change (P > 0.05) at the lowest frequency studied (0.2 Hz), but in some dogs it increased compared with control at the higher frequencies. Chest wall properties were not changed by emphysema (P > 0.05). We suggest that although large changes in acinar structure and EL occur during uncomplicated bilateral emphysema, secondary complications must be present to cause several of the characteristic dysfunctions seen in patients with emphysema. PMID- 9216963 TI - Fish oil and vitamin E supplementation in oxidative stress at rest and after physical exercise. AB - Fish oil supplementation and physical exercise may induce oxidative stress. We tested the effects of 8 wk of alpha-tocopherol (vitamin E) and fish oil (FO) supplementation on resting and exercise-induced oxidative stress. Rats (n = 80) were divided into groups supplemented with FO, FO and vitamin E (FOVE), soy oil (SO), and SO and vitamin E (SOVE), and for FOVE and SOVE they were divided into corresponding exercise groups (FOVE-Ex and SOVE-Ex). Lipid peroxidation [thiobarbituric acid-reacting substances (TBARS)] was 33% higher in FO compared with SO in the liver, but oxidative protein damage (carbonyl levels) remained similar in both liver and red gastrocnemius muscle (RG). Vitamin E supplementation, compared with FO and SO, markedly decreased liver and RG TBARS, but liver TBARS remained 32% higher in FOVE vs. SOVE. Vitamin E also markedly decreased liver and RG protein carbonyl levels, although levels in FOVE and SOVE were similar. Exercise increased liver and RG TBARS and RG protein carbonyl levels markedly, with similar levels in FOVE-Ex and SOVE-Ex. FO increased lipid peroxidation but not protein oxidation in a tissue-specific manner. Vitamin E markedly decreased lipid peroxidation and protein oxidation in both FOVE and SOVE, although liver lipid peroxidation remained higher in FOVE. Despite higher levels of hepatic lipid peroxidation at rest in FOVE compared with SOVE, liver appeared to be relatively less susceptible to exercise-induced oxidative stress in FOVE. PMID- 9216964 TI - Inputs from upper airway affect firing of respiratory-associated midbrain neurons. AB - In 16 decerebrated unanesthetized cats, we studied effects of neural inputs from upper airway on firing of 62 mesencephalic neurons that also developed respiratory-associated (RA) rhythmic firing when respiratory drive was high [Z. Chen, F. L. Eldridge, and P.G. Wagner. J. Physiol. (Lond.) 437: 305-325, 1991] and on firing of 16 neurons that did not develop the rhythmic firing (non-RA neurons). Activity in RA neurons increased after mechanical expansion of pharynx (45% of those tested) or larynx (68%) and after stimulation of glossopharyngeal (50%) or superior laryngeal nerves (77%). The increased neuronal firing occurred despite decreases or abolition of respiratory activity (expressed in phrenic nerve). Neuronal firing also increased after mechanical stimulation of nasal mucosa (66%) or by jets of air directed into the nares (48%) and after light brushing of nasal skin ( approximately 40%). Most stimuli led to decreased firing in a smaller number of neurons, and some neurons showed no response. None of the non-RA neurons developed an increase of firing after any of the stimuli, although one had decreased firing after stimulation of the superior laryngeal nerve. We conclude that inputs from the upper airway and nasal skin have independent modulatory effects on the same mesencephalic neurons that are stimulated by ascending rhythmic RA input from the medulla. These findings may have relevance to generation of the sensation of dyspnea. PMID- 9216965 TI - Absorption from different intestinal segments during exercise. AB - This study evaluated intestinal absorption from the first 75 cm of the proximal small intestine during 85 min of cycle exercise [63.6 +/- 0.7% peak O2 consumption (VO2 peak)] while subjects ingested either an isotonic carbohydrate electrolyte beverage (CHO-E) or a water placebo (WP). The CHO-E beverage contained 117 mM (4%) sucrose, 111 mM (2%) glucose, 18 meq Na+, and 3 meq K+. The two experiments were performed a week apart by seven subjects (6 men and 1 woman; mean VO2 peak = 53.5 +/- 6.5 ml . kg-1 . min-1). Nasogastric and multilumen tubes were fluoroscopically positioned in the gastric antrum and duodenojejunum, respectively. Subjects ingested 23 ml/kg body weight of the test solution, 20% (383 +/- 11 ml) of this volume 5 min before exercise and 10% (191 +/- 5 ml) every 10 min thereafter. By using the rate of gastric emptying (18.1 +/- 1.1 vs. 19.2 +/- 0. 7 ml/min for WP and CHO-E, respectively) as the rate of intestinal perfusion, intestinal absorption was determined by segmental perfusion from the duodenum (0-25 cm) and jejunum (25-50 and 50-75 cm). Water flux was different (P < 0.05) between solutions in the 0- to 25- and 25- to 50-cm segments for WP vs. CHO-E (30.7 +/- 2.7 vs. 15.0 +/- 2.9 and 3.8 +/- 1.1 vs. 11.9 +/- 3.3 ml . cm-1 . h-1, respectively). Furthermore, water flux differed (P < 0.05) for WP in a comparison of the 0- to 25- to the 25- to 50-cm segment. Total solute flux (TSF) was not significantly different among segments for a given solution or between solutions for a given segment. There was no difference between trials for percent change in plasma volume. These results indicate that 1) fluid absorption in the proximal small intestine depends on the segment studied and 2) solution composition can significantly effect water absorption rate in different intestinal segments. PMID- 9216966 TI - Postnatal lung function and protein permeability after fetal or maternal corticosteroids in preterm lambs. AB - We evaluated postnatal lung function and intravascular albumin loss to tissues of 123-days-gestation preterm surfactant-treated and ventilated lambs 15 h after direct fetal (n = 8) or maternal (n = 9) betamethasone treatment or saline placebo (n = 9). The betamethasone-treated groups had similar increases in dynamic compliances, ventilatory efficiency indexes, and lung volumes relative to controls (P < 0.05). The losses of 125I-labeled albumin from blood, a marker of intravascular integrity, and the recoveries of 125I-albumin in muscle and brain were similar for control and betamethasone-exposed lambs. Betamethasone-treated lambs had lower recoveries of 125I-albumin in lung tissues and in alveolar washes than did controls (P < 0.01). Although blood pressures were higher for the treated groups (P < 0.05), all groups had similar blood volumes, cardiac outputs, and organ blood flows. Maternal or fetal treatment with betamethasone 15 h before preterm delivery equivalently improved postnatal lung function, reduced albumin recoveries in lungs, and increased blood pressures. However, prenatal betamethasone had no effects on the systemic intravascular losses of albumin or did not change blood volumes. PMID- 9216967 TI - Sequential arousal and airway-defensive behavior of infants in asphyxial sleep environments. AB - Infants are prone to accidental asphyxiation. Therefore, we studied airway defensive behaviors and their relationship to spontaneous arousal behavior in 41 healthy sleeping infants (2-26 wk old), using two protocols: 1) infant was rebreathing expired air, face covered by bedding material; and 2) infant was exposed to hypercarbia, face uncovered. Multiple measurements of respiratory and motor activities were recorded (video, polygraph). The infants' response to increasing hypercarbia consisted of four highly stereotyped behaviors: sighs (augmented breaths), startles, thrashing limb movements, and full arousal (eyes open, cry). These behaviors occurred abruptly in self-limited clusters of activity and always in the same sequence: first a sigh coupled with a startle, then thrashing, then full arousal. Incomplete sequences (initial behaviors only) occurred far more frequently than the complete sequence and were variably effective in removing the bedding covering the airway. In both protocols, as inspired CO2 increased, incomplete arousal sequences recurred periodically and with increasing frequency and complexity until the infant either succeeded in clearing his/her airway or was completely aroused. Spontaneous arousal sequences, identical to those occurring during hypercarbia, occurred periodically during sleep. This observation suggests that the infant's airway-defensive responses to hypercarbia consist of an increase in the frequency and complexity of an endogenously regulated, periodically occurring sequence of arousal behaviors. PMID- 9216968 TI - Appendicular skeletal muscle mass: effects of age, gender, and ethnicity. AB - This study tested the hypothesis that skeletal muscle mass is reduced in elderly women and men after adjustment first for stature and body weight. The hypothesis was evaluated by estimating appendicular skeletal muscle mass with dual-energy X ray absorptiometry in a healthy adult cohort. A second purpose was to test the hypothesis that whole body 40K counting-derived total body potassium (TBK) is a reliable indirect measure of skeletal muscle mass. The independent effects on both appendicular skeletal muscle and TBK of gender (n = 148 women and 136 men) and ethnicity (n = 152 African-Americans and 132 Caucasians) were also explored. Main findings were 1) for both appendicular skeletal muscle mass (total, leg, and arm) and TBK, age was an independent determinant after adjustment first by stepwise multiple regression for stature and weight (multiple regression model r2 = approximately 0.60); absolute decrease with greater age in men was almost double that in women; significantly larger absolute amounts were observed in men and African-Americans after adjustment first for stature, weight, and age; and >80% of within-gender or -ethnic group between-individual component variation was explained by stature, weight, age, gender, and ethnicity differences; and 2) most of between-individual TBK variation could be explained by total appendicular skeletal muscle (r2 = 0.865), whereas age, gender, and ethnicity were small but significant additional covariates (total r2 = 0.903). Our study supports the hypotheses that skeletal muscle is reduced in the elderly and that TBK provides a reasonable indirect assessment of skeletal muscle mass. These findings provide a foundation for investigating skeletal muscle mass in a wide range of health related conditions. PMID- 9216969 TI - CO2 transport in normovolemic anemia: complete compensation and stability of blood CO2 tensions. AB - Isovolemic hemodilution does not appear to impair CO2 elimination nor cause CO2 retention despite the important role of red blood cells in blood CO2 transport. We studied this phenomenon and its physiological basis in eight New Zealand White rabbits that were anesthetized, paralyzed, and mechanically ventilated at a fixed minute ventilation. Isovolemic anemia was induced by simultaneous blood withdrawal and infusion of 6% hetastarch in sequential stages; exchange transfusions ranged from 15-30 ml in volume. Variables measured after each hemodilution included hematocrit (Hct), arterial and venous blood gases, mixed expired PCO2 and PO2, and blood pressure; also, O2 consumption, CO2 production, cardiac output (Q), and physiological dead space were calculated. Data were analyzed by comparison of changes in variables with changes in Hct and by using the model of capillary gas exchange described by Bidani (J. Appl. Physiol. 70: 1686-1699, 1991). There was complete compensation for anemia with stability of venous and arterial PCO2 between Hct values of 36 +/- 3 and 12 +/- 1%, which was predicted by the mathematical model. Over this range of hemodilution, Q rose 50%, and the O2 extraction ratio increased 61% without a decline in CO2 production or a rise in alveolar ventilation. The dominant compensations maintaining CO2 transport in normovolemic anemia include an increased Q and an augmented Haldane effect arising from the accompanying greater O2 extraction. PMID- 9216970 TI - When does the lung die? Kfc, cell viability, and adenine nucleotide changes in the circulation-arrested rat lung. AB - Lungs harvested from cadaveric circulation-arrested donors may increase the donor pool for lung transplantation. To determine the degree and time course of ischemia-reperfusion injury, we evaluated the effect of O2 ventilation on capillary permeability [capillary filtration coefficient (Kfc)], cell viability, and total adenine nucleotide (TAN) levels in in situ circulation-arrested rat lungs. Kfc increased with increasing postmortem ischemic time (r = 0.88). Lungs ventilated with O2 1 h postmortem had similar Kfc and wet-to-dry ratios as controls. Nonventilated lungs had threefold (P < 0.05) and sevenfold (P < 0.0001) increases in Kfc at 30 and 60 min postmortem compared with controls. Cell viability decreased in all groups except for 30-min postmortem O2-ventilated lungs. TAN levels decreased with increasing ischemic time, particularly in nonventilated lungs. Loss of adenine nucleotides correlated with increasing Kfc values (r = 0.76). This study indicates that lungs retrieved 1 h postmortem may have normal Kfc with preharvest O2 ventilation. The relationship between Kfc and TAN suggests that vascular permeability may be related to lung TAN levels. PMID- 9216971 TI - Ventilatory and metabolic responses to ambient hypoxia or hypercapnia in rats exposed to CO hypoxia. AB - We have investigated at ambient temperatures (Tam) of 25 and 5 degrees C the effects of ambient hypoxia (Hxam; fractional inspired O2 = 0.14) and hypercapnia (fractional inspired CO2 = 0.04) on ventilation (V), O2 uptake (VO2), and colonic temperature (Tc) in 12 conscious rats before and after carotid body denervation (CBD). The rats were concomitantly exposed to CO hypoxia (HxCO; fractional inspired CO = 0.03-0.05%), which decreases arterial O2 saturation by approximately 25-40%. The results demonstrate the following. 1) At Tam of 5 degrees C, in both intact and CBD rats, V/VO2 is larger when Hxam or CO2 is associated with HxCO than with normoxia. At Tam of 25 degrees C, this is also the case except for CO2 in CBD rats. 2) At Tam of 5 degrees C, the changes in VO2 and Tc seem to result from additive effects of the separate changes induced by Hxam, CO2, and HxCO. It is concluded that, in conscious rats, central hypoxia does not depress respiratory activity. On the contrary, particularly when VO2 is augmented during a cold stress, both V/VO2 during HxCO and the ventilatory responses to Hxam and CO2 are increased. The mechanisms involved in this relative hyperventilation are likely to involve diencephalic integrative structures. PMID- 9216972 TI - Anaerobic capacity and muscle activation during horizontal and uphill running. AB - Anaerobic capacity as measured by the maximal or peak oxygen deficit is greater during uphill than during horizontal running. The objective of this study was to determine whether the greater peak oxygen deficit determined during uphill compared with horizontal running is related to greater muscle volume or mass activated in the lower extremity. The peak oxygen deficit in 12 subjects was determined during supramaximal treadmill running at 0 and 10% grade. Exercise induced contrast shifts in magnetic resonance images were obtained before and after exercise and used to determine the percentage of muscle volume activated. The mean peak oxygen deficit determined for uphill running [2.96 +/- 0.63 (SD) liters or 49 +/- 6 ml/kg] was significantly greater (P < 0.05) than for horizontal running (2.45 +/- 0.51 liters or 41 +/- 7 ml/kg) by 21%. The mean percentage of muscle volume activated for uphill running [73.1 +/- 7. 4% (SD)] was significantly greater (P < 0.05) than for horizontal running (67.0 +/- 8.3%) by 9%. The differences in peak oxygen deficit (liters) between uphill and horizontal running were significantly related (y = 8.05 x 10(-4)x + 0.35; r = 0.63, SE of estimate = 0.29 liter, P < 0.05) to the differences in the active muscle volume (cm3) in the lower extremity. We conclude that the higher peak oxygen deficit during uphill compared with horizontal running is due in part to increased mass of skeletal muscle activated in the lower extremity. PMID- 9216973 TI - Physiological adaptations to a weight-loss dietary regimen and exercise programs in women. AB - Thirty-one women (mean age 35.4 +/- 8.5 yr) who were overweight were matched and randomly placed into either a control group (Con; n = 6), a diet-only group (D; n = 8), a diet+aerobic endurance exercise training group (DE; n = 9), or a diet+aerobic endurance exercise training+strength training group (DES; n = 8). After 12 wk, the three dietary groups demonstrated a significant (P 5% of the fibers sampled showed a hypertrophic response in both FO groups. One week of FO increased the percentage of hybrid (containing both type I and type IIa MHC) fibers and of fibers containing embryonic MHC. By 10 wk, the percentage of pure type I MHC fibers was approximately 40% in both FO groups compared with 15% in controls, and the percentage of fibers containing embryonic MHC was similar to that in controls. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analyses showed an increase in type I MHC and a decrease in type IIb MHC in both FO groups at 10 wk, whereas little change was observed at 1 wk. These data are consistent with hypertrophy and transformation from faster to slower MHC isoforms in chronically overloaded muscles. The additional overload imposed by daily endurance treadmill training employed in this study (1.6 km/day; 10% incline) results in a larger hypertrophic response but appears to have a minimal effect on the MHC adaptations. PMID- 9216976 TI - A simplified strong ion model for acid-base equilibria: application to horse plasma. AB - The Henderson-Hasselbalch equation and Stewart's strong ion model are currently used to describe mammalian acid-base equilibria. Anomalies exist when the Henderson-Hasselbalch equation is applied to plasma, whereas the strong ion model does not provide a practical method for determining the total plasma concentration of nonvolatile weak acids ([Atot]) and the effective dissociation constant for plasma weak acids (Ka). A simplified strong ion model, which was developed from the assumption that plasma ions act as strong ions, volatile buffer ions (HCO-3), or nonvolatile buffer ions, indicates that plasma pH is determined by five independent variables: PCO2, strong ion difference, concentration of individual nonvolatile plasma buffers (albumin, globulin, and phosphate), ionic strength, and temperature. The simplified strong ion model conveys on a fundamental level the mechanism for change in acid-base status, explains many of the anomalies when the Henderson-Hasselbalch equation is applied to plasma, is conceptually and algebraically simpler than Stewart's strong ion model, and provides a practical in vitro method for determining [Atot] and Ka of plasma. Application of the simplified strong ion model to CO2-tonometered horse plasma produced values for [Atot] (15.0 +/- 3.1 meq/l) and Ka (2.22 +/- 0.32 x 10(-7) eq/l) that were significantly different from the values commonly assumed for human plasma ([Atot] = 20.0 meq/l, Ka = 3.0 x 10(-7) eq/l). Moreover, application of the experimentally determined values for [Atot] and Ka to published data for the horse (known PCO2, strong ion difference, and plasma protein concentration) predicted plasma pH more accurately than the values for [Atot] and Ka commonly assumed for human plasma. Species-specific values for [Atot] and Ka should be experimentally determined when the simplified strong ion model (or strong ion model) is used to describe acid-base equilibria. PMID- 9216977 TI - Differences between estimates and measured PaCO2 during rest and exercise in older subjects. AB - Arterial PCO2 (PaCO2) has been estimated during exercise with good accuracy in younger individuals by using the Jones equation (PJCO2) (J. Appl. Physiol. 47: 954-960, 1979). The purpose of this project was to determine the utility of estimating PaCO2 from end-tidal PCO2 (PETCO2) or PJCO2 at rest, ventilatory threshold (VTh), and maximal exercise (Max) in older subjects. PETCO2 was determined from respired gases simultaneously (MGA 1100) with arterial blood gases (radial arterial catheter) in 12 older and 11 younger subjects at rest and during exercise. Mean differences were analyzed with paired t-tests, and relationships between the estimated PaCO2 values and the actual values of PaCO2 were determined with correlation coefficients. In the older subjects, PETCO2 was not significantly different from PaCO2 at rest (-1.2 +/- 4.3 Torr), VTh (0.4 +/- 2.5), or Max (-0.8 +/- 2.7), and the two were significantly (P < 0.05) correlated at Vth (r = 0.84) and Max (r = 0.87) but not at rest (r = 0.47). PJCO2 was similar to PaCO2 at rest (-1.0 +/- 3.9) and Vth (-1. 3 +/- 2.3) but significantly lower at Max (-3.0 +/- 2.6), and the two were significantly correlated at Vth (r = 0.86) and Max (r = 0. 80) but not at rest (r = 0.54). PETCO2 was significantly higher than PaCO2 during exercise in the younger subjects but similar to PaCO2 at rest. PJCO2 was similar to PaCO2 at rest and Vth but significantly lower at Max in younger subjects. In conclusion, our data demonstrate that PaCO2 during exercise is better estimated by PETCO2 than by PJCO2 in older subjects, contrary to what is observed in younger subjects. This appears to be related to the finding that PETCO2 does not exceed PaCO2 during exercise in older subjects, as occurs in the younger subjects. However, PaCO2 at rest is best estimated by PJCO2 in both younger and older subjects. PMID- 9216978 TI - Spiral nerve cuff electrode for recordings of respiratory output. AB - The feasibility of using the spiral nerve cuff electrode design for recordings of respiratory output from the hypoglossal (HG) and phrenic nerves is demonstrated in anesthetized, paralyzed, and artificially ventilated cats. Raw neural discharges of the HG nerve were analyzed in terms of signal-to-noise ratios and frequency spectra. The rectified and integrated moving average activity of the HG nerve had a peak value of 1.74 +/- 0.21 microV and a baseline value of 0.72 +/- 0.11 microV at elevated respiratory drive induced by increases in CO2 or oxygen deprivation when recorded with 10-mm-long cuffs. The frequency content of the HG electroneurogram extended from several hundred hertz to 6 kHz. Spiral nerve cuff recordings without desheathing of the nerve provided large enough signal-to-noise ratios that allowed them to be used as a measure of respiratory output and had much wider frequency bandwidths than the hook electrode preparations. A major advantage of the cuff electrode over the hook electrode was its mechanical stability, which significantly improved the reproducibility of the recordings both in terms of signal amplitudes and frequency contents. PMID- 9216979 TI - Decreased [3H]ouabain binding sites in skeletal muscle of rats with chronic heart failure. AB - Abnormalities intrinsic to skeletal muscle are thought to contribute to decrements in exercise capacity found in individuals with chronic heart failure (CHF). Na+-K+-adenosinetriphosphatase (the Na+ pump) is essential for maintaining muscle excitability and contractility. Therefore, we investigated the possibility that the number and affinity of Na+ pumps in locomotor muscles of rats with CHF are decreased. Myocardial infarction (MI) was induced in 8 rats, and a sham operation was performed in 12 rats. The degree of CHF was assessed approximately 180 days after surgery. Soleus and plantaris muscles were harvested, and Na+ pumps were quantified by using a [3H]ouabain binding assay. At the time of muscle harvest, MI and sham-operated rats were similar in age (458 +/- 54 vs. 447 +/- 34 days old, respectively). Compared with their sham-operated counterparts, MI rats had a significant amount of heart failure, right ventricular-to-body weight ratio was greater (48%), and the presence of pulmonary congestion was suggested by an elevated lung-to-body weight ratio (29%). Left ventricular end-diastolic pressure was significantly increased in the MI rats (11 +/- 1 mmHg) compared with the sham operated controls (1 +/- 1 mmHg). In addition, mean arterial blood pressure was lower in the MI rats compared with their control counterparts. [3H]ouabain binding sites were reduced 18% in soleus muscle (136 +/- 12 vs. 175 +/- 13 pmol/g wet wt, MI vs. sham, respectively) and 22% in plantaris muscle (119 +/- 12 vs. 147 +/- 8 pmol/g wet wt, MI vs. sham, respectively). The affinity of these [3H]ouabain binding sites was similar for the two groups. The relationship between the reduction in Na+ pump number and the reduced exercise capacity in individuals with CHF remains to be determined. PMID- 9216981 TI - Motor neurone disease and animal models. PMID- 9216982 TI - APP gene family: unique age-associated changes in splicing of Alzheimer's betaA4 amyloid protein precursor. AB - The betaA4-amyloid protein precursor (APP) is a transmembrane glycoprotein that is the source of the characteristic betaA4-amyloid deposits of Alzheimer brains. It exists in eight isoforms generated by alternative splicing of exons 7, 8 and 15, of which the L-APP mRNAs lacking exon 15 are significantly expressed in non neuronal cells and tissues, but not in neurones. Recently, it was shown that APP is a member of a multigene family of which the amyloid precursor-like protein 2 (APLP2) is the nearest relative. Analysis of APLP2 expression revealed regulated alternative splicing of the Kunitz protease inhibitor domain (KPI, homologous to exon 7 of APP) and a non-homologous insert of 12 amino acids on the NH2-terminal side of the transmembrane domain. While expression of the KPI encoding exon of APLP2 is abundant in neurones and thus differs from APP, L-APLP2 mRNA isoforms lacking the latter, non-homologous insert show a tissue-specific expression pattern similar to that of exon 15 of APP. Comparison of alternatively spliced APP and APLP2 mRNA isoforms in rat brain regions from early post-natal and adult rats revealed significantly higher relative amounts of KPI-encoding APP isoforms in the adult rat brain and an even more pronounced augmentation of L-APP mRNAs. Both effects were not observed for APLP2. This indicates an APP-specific age associated regulation pattern within the APP gene family which has intriguing implications for the development of Alzheimer's disease in humans. PMID- 9216983 TI - Scrapie infection of transgenic mice leads to network and intrinsic dysfunction of cortical and hippocampal neurones. AB - The human prion encephalopathy Creutzfeldt-Jakob disease often is manifest as rapidly progressing dementia with myoclonus and synchronous, periodic discharges. To investigate the electrophysiology of prion disease we used intra- and extra cellular recordings from brain slices from Tg(SHaPrP+/+) 81 mice, which express Syrian hamster prion protein and which are susceptible to hamster-passaged scrapie isolates. Forty days after intracerebral inoculation with scrapie isolate Sc237, we recorded prolonged, epileptiform discharges in cortex and hippocampus. Neurological signs were subtle and histopathology was minimal. Central nervous system (CNS) dysfunction progressed; by 57 days the mice were ataxic, had spongiform histopathology and they died in <63 days. During the terminal phase, intrinsic neuronal properties changed dramatically and action potentials broadened from <4 to 20-100 ms in 30% of cortical cells. We conclude that brain dysfunction in experimental scrapie precedes clinical signs and spongiform histopathology, and is preserved in slices maintained in vitro, making it accessible to electrophysiological analysis. PMID- 9216984 TI - Induced spreading depressions in energy-compromised neocortical tissue: calcium transients and histopathological correlates. AB - Mechanisms causing gradual recruitment of damaged cells in the penumbra zone around the core of a focal ischaemic lesion may encompass irregularly occurring depolarization waves of the spreading depression (SD) type, each leading to transient loading of cells with calcium. It has been speculated that, when elicited in an underperfused or otherwise energy-compromised tissue, such depolarization waves lead to cell damage. We assessed under what conditions the calcium transients during KCl-induced SDs are prolonged, and explored if marked prolongation of the transients leads to brain damage. Cerebral blood flow (CBF) was reduced by marked hypocapnia. Tissue oxygenation was reduced by arterial hypoxia, without or with unilateral carotid artery occlusion, or by occlusion of the carotid arteries in normoxic, anaesthetized rats. In all animals the DC potential and extracellular calcium concentration (Ca2+e) were measured before and during a series of SDs. The animals were recovered for histopathological assessment. Hypoxia alone (Pao2, 32.5 +/- 3.8 mmHg) increased mean and total depolarization times, but repeated SDs elicited over 1.7 (+/-0.4) h failed to induce cell damage. Unilateral carotid artery occlusion further prolonged the SD waves but, in spite of total depolarization times of up to 40 min during 2 h, only two out of seven animals showed damage, localized to caudoputamen and parietal cortex, as well as to the subiculum, CA1 and CA3 sectors of the hippocampus. Bilateral carotid artery occlusion was associated with the most pronounced prolongation of the DC potential shifts and Ca2+ transients, with total depolarization times of up to 70 min. In spite of this, only four out of 13 animals showed brain damage and in two of these the damage was contralateral. The results justify modification of the hypothesis stating that SD-like depolarizations in the perifocal penumbra zone per se is what leads to gradual recruitment of such tissues in the infarction process. It is suggested that additional factors are required, such as a larger reduction in CBF, or the proximity of cells at risk to necrotic tissue. PMID- 9216985 TI - Cortical neurones with Ca2+ permeable AMPA/kainate channels display distinct receptor immunoreactivity and are GABAergic. AB - A minority subset of cortical neurones exhibit kainate-activated Co2+ uptake, a marker for AMPA/kainate receptor gated Ca2+-permeable channels. Consistent with enhanced Ca2+ influx through these channels, Co2+-positive neurones are unusually vulnerable to death induced by exposure to either AMPA or kainate. Here we show that Co2+-positive cortical neurones express a distinctive profile of AMPA receptor subunits as determined by immunostaining. Co2+-positive neurones were much less likely to express GluR2/GluR3, and much more likely to express GluR1 or GluR4, than the general cortical neuronal population. Thus expression of AMPA receptors lacking the GluR2 subunit may explain the Co2+ staining, and selective vulnerability to kainate exhibited by Co2+-positive cells. Almost all GABAergic neurones, identified by immunostaining for glutamic acid decarboxylase, were Co2+ positive. The widespread presence of Ca2+-permeable AMPA/kainate receptor-gated channels on cortical GABAergic neurones may have important implications for the fate of cortical inhibition in disease states associated with the excitotoxic overstimulation of glutamate receptors. PMID- 9216986 TI - NOS induction by NGF in basal forebrain cholinergic neurones: evidence for regulation of brain NOS by a neurotrophin. AB - Nerve growth factor (NGF) acts through trkA receptors to serve as a trophic factor for cholinergic neurones in the medial septal nucleus (MSN) and vertical limb of the diagonal band (VDB). Herein, we show that brain nitric oxide synthase (NOS), which synthesizes the neuromodulator nitric oxide, is selectively expressed in a large fraction of trkA-containing neurones in the MSN and VDB. Axotomy of these neurones gave evidence that NOS expressing cholinergic neurones innervate the hippocampus. NGF infusion induced a robust, specific increase in NOS expression in basal forebrain cholinergic neurones. These results indicate that brain NOS can be regulated by a neurotrophic factor and suggest that NGF influences forebrain function by regulating production of nitric oxide as well as acetylcholine. PMID- 9216987 TI - Identification of new human myelin basic protein transcripts in the immune and central nervous systems. AB - Five new myelin basic protein (MBP) transcripts were identified which each have preferential sites of expression in adult human brain and immune system. They contain a novel 5' coding region which presents extensive sequence similarity to the mouse 0' region. One of these ribonucleic acid (RNA) species, HMBPR1, is found essentially, if not only, in haemopoietic and immune cells. Two alternatively spliced transcripts called MBP2a and c are only expressed in the central nervous system (CNS). In addition, the two other transcripts are expressed in both immune and nervous systems. Thus, the MBP locus can generate multiple forms of RNA, whose start sites and splicing depend on the tissue in which they are expressed. The presence of an MBP transcript specifically expressed in the adult human immune system suggests previously unsuspected functions related to the pathogenesis of multiple sclerosis. PMID- 9216988 TI - Somatic gene transfer to the adult primate central nervous system: in vitro and in vivo characterization of cells genetically modified to secrete nerve growth factor. AB - Somatic gene transfer offers a means of delivering substances to the central nervous system (CNS) in a regionally specific, high-dose, chronic and well tolerated manner. Studies in rats have shown that genetically modified cell grafts can prevent neuronal degeneration and promote functional recovery after injury and can improve cognitive function in aged subjects. To assess the potential utility of somatic gene transfer techniques in primate models, retroviral vectors were used to modify genetically monkey and human primary skin fibroblasts to produce and secrete human nerve growth factor (NGF). In vitro, all cell types produced NGF and sustained this production through cell growth to confluency, as determined by both Northern blot analysis and ELISA. Adult human fibroblasts produced as much NGF as did young human fibroblasts. Monkey fibroblasts genetically modified to produce NGF were then grafted to intact adult rhesus and cynomolgous monkey brains. Among nine primates that received a total of 76 grafts, surviving cells were found in all subjects up to the maximal experimental timepoint of 6 months. Cholinergic fibres from the host brain penetrated NGF-secreting grafts up to 6 months after grafting, but showed little penetration in control grafts lacking the NGF gene. Autografts survived better than allografts. These findings indicate that both human and primate fibroblasts can be transduced to produce and secrete NGF, can maintain this production whether in a growing or quiescent state and can elicit robust sprouting responses when primate fibroblasts are grafted to the adult brain. Cells genetically modified to produce trophic factors are a useful model for studying in vitro and in vivo CNS plasticity and for testing potential therapies for neurodegenerative conditions. PMID- 9216989 TI - The gene for Machado-Joseph disease maps to the same 3-cM interval as the spinal cerebellar ataxia 3 gene on chromosome 14q. AB - Machado-Joseph disease (MJD) is an autosomal dominant neurodegenerative disorder in families of Portuguese-Azorean ancestry. The gene responsible for MJD has been assigned to a 29-cM interval on chromosome 14q. A large Brazilian family with MJD was genotyped with six new microsatellite markers spanning 19 cM on chromosome 14q. Linkage analysis and haplotype reconstruction reduced the MJD candidate region to a 3-cM interval between markers D14S280 and D14S81, permitting positional cloning. This interval also contains the spinal cerebellar ataxia 3 (SCA3) gene, responsible for a genetic subtype of the type I autosomal dominant cerebellar ataxias, clinically related to MJD. This result supports the hypothesis that abnormalities in the same gene may be responsible for both disorders. The minor clinical differences between the two diseases may result from allelic heterogeneity. PMID- 9216990 TI - Infection of sympathetic and sensory neurones with herpes simplex virus does not elicit a shut-off of cellular protein synthesis: implications for viral latency and herpes vectors. AB - Infection of non-neuronal cell types with herpes simplex virus type 1 (HSV-1) results in the degradation of host mRNA (Kwong & Frenkel 1987) and a shutoff in host protein synthesis (Roizman et al. 1965). This effect is mediated by a virion associated protein that is encoded by the viral vhs gene (Read & Frenkel 1983). This virion host shutoff (VHS) helps regulate viral gene expression and promotes efficient viral replication during the lytic cycle (Kwong & Frenkel 1987). Cultured sympathetic and sensory neurones, in contrast to primary rat fibroblasts, PC-12 cells, and Vero cells, showed no reduction in protein synthesis following infection with HSV-1. The resistance of neurones to VHS may be important in allowing establishment of a latent infection. In addition, this finding has a favourable impact on the idea of using HSV as a vector to deliver foreign genes into neurones. PMID- 9216991 TI - A nonsense mutation in the alpha4 subunit of the nicotinic acetylcholine receptor (CHRNA4) cosegregates with 20q-linked benign neonatal familial convulsions (EBNI) AB - Benign Familial Neonatal Convulsions (BFNC) is an epileptic disorder with an autosomal dominant mode of transmission. It has been shown that about 80% of BFNC pedigrees are linked to a genetic defect on chromosome 20q13.3. A candidate gene for the epilepsies, the gene coding for the alpha4 subunit of the nicotinic cholinergic receptor (CHRNA4), has previously been localized on chromosome 20. Here we report a single point mutation converting a serine codon to a stop codon in the exon 5 of CHRNA4, in one BFNC family. Identification of CHRNA4 as the defective gene in 20q-BFNC represents the first example of a human idiopathic epilepsy caused by a mutation directly affecting a neurotransmitter receptor in the central nervous system. PMID- 9216992 TI - A role for ultraspiracle, the Drosophila RXR, in morphogenetic furrow movement and photoreceptor cluster formation. AB - Many of the same genes needed for proper eye and limb development in vertebrates, such as hairy, hedgehog, patched and cyclic AMP-dependent protein kinase A, are responsible for patterning Drosophila imaginal discs, the tissues that will give rise to the adult cuticle structures. This is well demonstrated in the control of morphogenetic furrow movement and differentiation in the eye imaginal disc. We report that ultraspiracle, the gene encoding the Drosophila cognate of the Retinoid X Receptor, is required for normal morphogenetic furrow movement and ommatidial cluster formation. Examination of the expression of genes involved in regulating the furrow suggests that ultraspiracle defines a novel regulatory pathway in eye differentiation. PMID- 9216993 TI - Myogenin can substitute for Myf5 in promoting myogenesis but less efficiently. AB - The myogenic basic Helix-Loop-Helix transcription factors, including Myf5, MyoD, myogenin (myg) and MRF4, play important roles in skeletal muscle development. The phenotypes of mutant mice deficient in either gene are different, suggesting that each gene may have a unique function in vivo. We previously showed that targeting myogenin into the Myf5 locus (Myf5(myg-ki)) rescued the rib cage truncation in the Myf5-null mutant, hence demonstrating functional redundancy between Myf5 and myogenin in skeletal morphogenesis. Here we present the results of crossing myogenin knock-in (myg-ki) mice with either MyoD-null or myogenin-null mutants. The Myf5(myg-ki) allele rescued early myogenesis, but Myf5(myg-ki/myg-ki);MyoD(-/ ) mutant mice died immediately after birth owing to reduced muscle formation. Therefore, myogenin, expressed from the Myf5 locus, is not able to completely replace the function of Myf5 in muscle development although it is capable of determining and/or maintaining myogenic lineage. Myf5(myg-ki/myg-ki);myg(-/-) mutant mice displayed the same phenotype as myg(-/-) mutants. This indicates that the earlier expression of myogenin cannot promote myogenic terminal differentiation, which is normally initiated by the endogenous myogenin. Thus, our results are consistent with the notion that Myf5 and myogenin are functionally interchangeable in determining myogenic lineage and assuring normal rib formation. Our experiment revealed, however, that some aspects of myogenesis may be unique to a given myogenic factor and are due to either different regulatory sequences that control their temporal and spatial expression or different functional protein domains. PMID- 9216994 TI - The differentiation of the serotonergic neurons in the Drosophila ventral nerve cord depends on the combined function of the zinc finger proteins Eagle and Huckebein. AB - The Drosophila ventral nerve cord (vNC) derives from a stereotyped population of neural stem cells, neuroblasts (NBs), each of which gives rise to a characteristic cell lineage. The mechanisms leading to the specification and differentiation of these lineages are largely unknown. Here we analyse mechanisms leading to cell differentiation within the NB 7-3 lineage. Analogous to the grasshopper, NB 7-3 is the progenitor of the Drosophila vNC serotonergic neurons. The zinc finger protein Eagle (Eg) is expressed in NB 7-3 just after delamination and is present in all NB 7-3 progeny until late stage 17. DiI cell lineage tracing and immunocytochemistry reveal that eg is required for normal pathfinding of interneuronal projections and for restricting the cell number in the thoracic NB 7-3 lineage. Moreover, eg is required for serotonin expression. Ectopic expression of Eg protein forces specific additional CNS cells to enter the serotonergic differentiation pathway. Like NB 7-3, the progenitor(s) of these ectopic cells express Huckebein (Hkb), another zinc finger protein. However, their progenitors do not express engrailed (en) as opposed to the NB 7-3 lineage, where en acts upstream of eg. We conclude that eg and hkb act in concert to determine serotonergic cell fate, while en is more distantly involved in this process by activating eg expression. Thus, we provide the first functional evidence for a combinatorial code of transcription factors acting early but downstream of segment polarity genes to specify a unique neuronal cell fate. PMID- 9216995 TI - beta3-tubulin is directly repressed by the engrailed protein in Drosophila. AB - In Drosophila, Engrailed is a nuclear regulatory protein with essential roles during embryonic development. Although Engrailed is a transcription factor, little progress has been achieved in identifying its target genes. We report here the identification of an effector gene, the beta3-tubulin gene, as a direct target of Engrailed. The cytological location of beta3-tubulin, 60C, is a strong site of Engrailed binding on polytene chromosomes. Immunostaining analysis of a transgenic line containing a P[beta3-tubulin-lacZ] construct shows an additional site of Engrailed binding at the location of the transgene. Molecular analysis allowed identification of several Engrailed binding sites, both in vitro and in vivo, within the first intron of the beta3-tubulin locus. Engrailed binding sites identified in vitro are active in larvae. Furthermore, expression of beta3 tubulin is derepressed in the ectoderm of engrailed mutant embryos. Repression of beta3-tubulin by Engrailed is also obtained when Engrailed is ectopically expressed in embryonic mesoderm. Finally, two different sets of Engrailed binding sites are shown to be involved in the early and late regulation of beta3-tubulin by Engrailed during embryogenesis. PMID- 9216996 TI - Gli1 is a target of Sonic hedgehog that induces ventral neural tube development. AB - The vertebrate zinc finger genes of the Gli family are homologs of the Drosophila gene cubitus interruptus. In frog embryos, Gli1 is expressed transiently in the prospective floor plate during gastrulation and in cells lateral to the midline during late gastrula and neurula stages. In contrast, Gli2 and Gli3 are absent from the neural plate midline with Gli2 expressed widely and Gli3 in a graded fashion with highest levels in lateral regions. In mouse embryos, the three Gli genes show a similar pattern of expression in the neural tube but are coexpressed throughout the early neural plate. Because Gli1 is the only Gli gene expressed in prospective floor plate cells of frog embryos, we have investigated a possible involvement of this gene in ventral neural tube development. Here we show that Shh signaling activates Gli1 transcription and that widespread expression of endogenous frog or human glioma Gli1, but not Gli3, in developing frog embryos results in the ectopic differentiation of floor plate cells and ventral neurons within the neural tube. Floor-plate-inducing ability is retained when cytoplasmic Gli1 proteins are forced into the nucleus or are fused to the VP16 transactivating domain. Thus, our results identify Gli1 as a midline target of Shh and suggest that it mediates the induction of floor plate cells and ventral neurons by Shh acting as a transcriptional regulator. PMID- 9216997 TI - Xmsx-1 modifies mesodermal tissue pattern along dorsoventral axis in Xenopus laevis embryo. AB - This study analyzes the expression and the function of Xenopus msx-1 (Xmsx-1) in embryos, in relation to the ventralizing activity of bone morphogenetic protein-4 (BMP-4). Expression of Xmsx-1 was increased in UV-treated ventralized embryos and decreased in LiCl-treated dorsalized embryos at the neurula stage (stage 14). Whole-mount in situ hybridization analysis showed that Xmsx-1 is expressed in marginal zone and animal pole areas, laterally and ventrally, but not dorsally, at mid-gastrula (stage 11) and late-gastrula (stage 13) stages. Injection of BMP 4 RNA, but not activin RNA, induced Xmsx-1 expression in the dorsal marginal zone at the early gastrula stage (stage 10+), and introduction of a dominant negative form of BMP-4 receptor RNA suppressed Xmsx-1 expression in animal cap and ventral marginal zone explants at stage 14. Thus, Xmsx-1 is a target gene specifically regulated by BMP-4 signaling. Embryos injected with Xmsx-1 RNA in dorsal blastomeres at the 4-cell stage exhibited a ventralized phenotype, with microcephaly and swollen abdomen. Histological observation and immunostaining revealed that these embryos had a large block of muscle tissue in the dorsal mesodermal area instead of notochord. On the basis of molecular marker analysis, however, the injection of Xmsx-1 RNA did not induce the expression of alpha globin, nor reduce cardiac alpha-actin in dorsal marginal zone explants. Furthermore, a significant amount of alpha-actin was induced and alpha-globin was turned off in the ventral marginal zone explants injected with Xmsx-1. These results indicated that Xmsx-1 is a target gene of BMP-4 signaling, but possesses a distinct activity on dorsal-ventral patterning of mesodermal tissues. PMID- 9216998 TI - Induction of cardiac myogenesis in avian pregastrula epiblast: the role of the hypoblast and activin. AB - An in vitro assay has been developed to investigate tissue interactions regulating myocardial cell specification in birds. Explants from the posterior region of stage XI-XIV blastulas were found to form heart muscle at high frequency with a timing that corresponded to onset of cardiac myocyte differentiation in vivo. Isolation and recombination experiments demonstrated that a signal from the hypoblast was required to induce cardiac myogenesis in the epiblast, and regional differences in epiblast responsiveness and hypoblast inductiveness restrict appearance of cardiac myocytes to the posterior region. Explantation studies provided evidence that myocardial cell specification is underway by stage 3, indicating that the hypoblast-derived signal occurs shortly before specification is detected. Recombinations were also performed to compare cardiac-inducing capacities of pregastrula hypoblast and stage 5 anterior lateral endoderm. The hypoblast possessed broad capacity to induce heart muscle cells in pregastrula and mid-gastrula epiblast, and modest ability to induce cardiac myogenesis in stage 4 posterior primitive streak. Stage 5 anterior lateral endoderm, in contrast, showed no ability to induce heart development in epiblast cells but was a potent inducer of cardiac myogenesis in cells from stage 4 posterior primitive streak. These findings suggest that the hypoblast-derived signal likely acts upstream of proposed heart-inducing signals provided by anterior lateral endoderm. Experiments were also performed to investigate whether activin, or an activin-like molecule, is involved in regulating cardiac myogenesis. Follistatin blocked cardiac myogenesis in stage XI-XIV posterior region explants and activin induced cardiac myogenesis in a dose-dependent fashion in posterior epiblast. These findings indicate that activin, or an activin-like molecule, is required for and is sufficient to stimulate cardiac myogenesis in posterior region pregastrula epiblast. Three models are presented to explain these results. PMID- 9216999 TI - Genes that guide growth cones along the C. elegans ventral nerve cord. AB - During nervous system development, growth cone pioneering and fasciculation contribute to nerve bundle structure. Pioneer growth cones initially navigate along neuroglia to establish an axon scaffold that guides later extending growth cones. In C. elegans, the growth cone of the PVPR neuron pioneers the left ventral nerve cord bundle, providing a path for the embryonic extensions of the PVQL and AVKR growth cones. Later during larval development, the HSNL growth cone follows cues in the left ventral nerve cord bundle provided by the PVPR and PVQL axons. Here we show that mutations in the genes enu-1, fax-1, unc-3, unc-30, unc 42 and unc-115 disrupt pathfinding of growth cones along the left ventral nerve cord bundle. Our results indicate that unc-3 and unc-30 function in ventral nerve cord pioneering and that enu-1, fax-1, unc-42 and unc-115 function in recognition of the PVPR and PVQL axons by the AVKR and HSNL growth cones. PMID- 9217000 TI - A role for Siamois in Spemann organizer formation. AB - The vertebrate body plan is specified in the early embryo through the inductive influence of the organizer, a special region that forms on the dorsalmost side of the embryo at the beginning of gastrulation. In Xenopus, the homeobox gene Siamois is activated prior to gastrulation in the area of organizer activity and is capable of inducing a secondary body axis when ectopically expressed. To elucidate the function of endogeneous Siamois in dorsoventral axis formation, we made a dominant repressor construct (SE) in which the Siamois homeodomain was fused to an active repression domain of Drosophila engrailed. Overexpression of 1 5 pg of this chimeric mRNA in the early embryo blocks axis development and inhibits activation of dorsal, but not ventrolateral, marginal zone markers. At similar expression levels, SE proteins with altered DNA-binding specificity do not have the same effect. Coexpression of mRNA encoding wild-type Siamois, but not a mutated Siamois, restores dorsal development to SE embryos. Furthermore, SE strongly blocks axis formation triggered by beta-catenin but not by the organizer product noggin. These results suggest that Siamois function is essential for beta catenin-mediated formation of the Spemann organizer, and that Siamois acts prior to noggin in specifying dorsal development. PMID- 9217001 TI - The Drosophila G-protein-coupled receptor kinase homologue Gprk2 is required for egg morphogenesis. AB - G protein signaling is a widely utilized form of extracellular communication that is mediated by a family of serpentine receptors containing seven transmembrane domains. In sensory neurons, cardiac muscle and other tissues, G protein-coupled receptors are desensitized through phosphorylation by a family of kinases, the G protein-coupled receptor kinases (GRKs). Desensitization allows a cell to decrease its response to a given signal, in the continued presence of that signal. We have identified a Drosophila mutant, gprk2(6936) that disrupts expression of a putative member of the GRK family, the G protein-coupled receptor kinase 2 gene (Gprk2). This mutation affects Gprk2 gene expression in the ovaries and renders mutant females sterile. The mutant eggs contain defects in several anterior eggshell structures that are produced by specific subsets of migratory follicle cells. In addition, rare eggs that become fertilized display gross defects in embryogenesis. These observations suggest that developmental signals transduced by G protein-coupled receptors are regulated by receptor phosphorylation. Based on the known functions of G protein-coupled receptor kinases, we speculate that receptor desensitization assists cells that are migrating or undergoing shape changes to respond rapidly to changing external signals. PMID- 9217002 TI - Limb proprioceptive deficits without neuronal loss in transgenic mice overexpressing neurotrophin-3 in the developing nervous system. AB - The role of neurotrophin-3 (NT3) during sensory neuron development was investigated in transgenic mice overexpressing NT3 under the control of the promoter and enhancer regions of the nestin gene, an intermediate filament gene widely expressed in the developing nervous system. Most of these mice died during the first postnatal day, and all showed severe limb ataxia suggestive of limb proprioceptive dysfunction. Tracing and histological analyses revealed a complete loss of spindles in limb muscles, absence of peripheral and central Ia projections, and lack of cells immunoreactive to parvalbumin in the dorsal root ganglion (DRG). Despite these deficits, there was no neuronal loss in the DRG of these mice. At birth, transgenic DRG showed increased neuron numbers, and displayed a normal proportion of neurons expressing substance P, calcitonin gene related peptide and the NT3 receptor trkC. Transgenic dorsal roots exhibited an increased number of axons at birth, indicating that all sensory neurons in transgenic mice projected to the dorsal spinal cord. Despite the absence of central Ia afferents reaching motorneurons, several sensory fibers were seen projecting towards ectopic high levels of NT3 in the midline of transgenic spinal cords. These findings suggest novel roles for NT3 in differentiation of proprioceptive neurons, target invasion and formation of Ia projections which are independent from its effects on neuronal survival. PMID- 9217003 TI - Reciprocal signaling between Drosophila epidermal muscle attachment cells and their corresponding muscles. AB - Directed intercellular interactions between distinct cell types underlie the basis for organogenesis during embryonic development. This paper focuses on the establishment of the final somatic muscle pattern in Drosophila, and on the possible cross-talk between the myotubes and the epidermal muscle attachment cells, occurring while both cell types undergo distinct developmental programs. Our findings suggest that the stripe gene is necessary and sufficient to initiate the developmental program of epidermal muscle attachment cells. In stripe mutant embryos, these cells do not differentiate correctly. Ectopic expression of Stripe in various epidermal cells transforms these cells into muscle-attachment cells expressing an array of epidermal muscle attachment cell-specific markers. Moreover, these ectopic epidermal muscle attachment cells are capable of attracting somatic myotubes from a limited distance, providing that the myotube has not yet been attached to or been influenced by a closer wild-type attachment cell. Analysis of the relationships between muscle binding and differentiation of the epidermal muscle attachment cell was performed in mutant embryos in which loss of muscles, or ectopic muscles were induced. This analysis indicated that, although the initial expression of epidermal muscle-attachment cell-specific genes including stripe and groovin is muscle independent, their continuous expression is maintained only in epidermal muscle attachment cells that are connected to muscles. These results suggest that the binding of a somatic muscle to an epidermal muscle attachment cell triggers a signal affecting gene expression in the attachment cell. Taken together, our results suggest the presence of a reciprocal signaling mechanism between the approaching muscles and the epidermal muscle attachment cells. First the epidermal muscle attachment cells signal the myotubes and induce myotube attraction and adhesion to their target cells. Following this binding, the muscle cells send a reciprocal signal to the epidermal muscle attachment cells inducing their terminal differentiation into tendon-like cells. PMID- 9217004 TI - Genetic evidence that heparin-like glycosaminoglycans are involved in wingless signaling. AB - We have identified the Drosophila UDP-glucose dehydrogenase gene as being involved in wingless signaling. Mutations in this gene, called kiwi, generate a phenotype identical to that of wingless. UDP-glucose dehydrogenase is required for the biosynthesis of UDP-glucuronate, which in turn is utilized in the biosynthesis of glycosaminoglycans. By rescuing the kiwi phenotype with both UDP glucuronate and the glycosaminoglycan heparan sulfate, we show that kiwi function in the embryo is crucial for the production of heparan sulfate in the extracellular matrix. Further, injection of heparin degrading enzyme, heparinase (and not chondroitin, dermatan or hyaluronic acid degrading enzyme) into wild type embryos leads to the degradation of heparin-like glycosaminoglycans and a 'wingless-like' cuticular phenotype. Our study thus provides the first genetic evidence for the involvement of heparin-like glycosaminoglycans in signal transduction. PMID- 9217005 TI - Development of branchiomotor neurons in zebrafish. AB - The mechanisms underlying neuronal specification and axonogenesis in the vertebrate hindbrain are poorly understood. To address these questions, we have employed anatomical methods and mutational analysis to characterize the branchiomotor neurons in the zebrafish embryo. The zebrafish branchiomotor system is similar to those in the chick and mouse, except for the location of the nVII and nIX branchiomotor neurons. Developmental analyses of genes expressed by branchiomotor neurons suggest that the different location of the nVII neurons in the zebrafish may result from cell migration. To gain insight into the mechanisms underlying the organization and axonogenesis of these neurons, we examined the development of the branchiomotor pathways in neuronal mutants. The valentino b337 mutation blocks the formation of rhombomeres 5 and 6, and severely affects the development of the nVII and nIX motor nuclei. The cyclops b16 mutation deletes ventral midline cells in the neural tube, and leads to a severe disruption of most branchiomotor nuclei and axon pathways. These results demonstrate that rhombomere-specific cues and ventral midline cells play important roles in the development of the branchiomotor pathways. PMID- 9217006 TI - TETRASPORE is required for male meiotic cytokinesis in Arabidopsis thaliana. AB - In flowering plants, male meiosis occurs in the microsporocyte to produce four microspores, each of which develops into a pollen grain. Here we describe four mutant alleles of TETRASPORE (TES), a gene essential for microsporocyte cytokinesis in Arabidopsis thaliana. Following failure of male meiotic cytokinesis in tes mutants, all four microspore nuclei remain within the same cytoplasm, with some completing their developmental programmes to form functional pollen nuclei. Both of the mitotic divisions seen in normal pollen development take place in tes mutants, including the asymmetric division required for the differentiation of gametes; some tes grains perform multiple asymmetric divisions in the same cytoplasm. tes pollen shows a variety of abnormalities subsequent to the cytokinetic defect, including fusion of nuclei, formation of ectopic internal walls, and disruptions to external wall patterning. In addition, ovules fertilized by tes pollen often abort, possibly because of excess paternal genomes in the endosperm. Thus tes mutants not only reveal a gene specific to male meiosis, but aid investigation of a wide range of processes in pollen development and function. PMID- 9217008 TI - Evolution of phosphagen kinase V. cDNA-derived amino acid sequences of two molluscan arginine kinases from the chiton Liolophura japonica and the turbanshell Battilus cornutus. AB - The cDNAs of arginine kinases from the chiton Liolophura japonica (Polyplacophora) and the turbanshell Battilus cornutus (Gastropoda) were amplified by polymerase chain reaction (PCR), and the complete nucleotide sequences of 1669 and 1624 bp, respectively, were determined. The open reading frame for Liolophura arginine kinase is 1050 nucleotides in length and encodes a protein with 349 amino acid residues, and that for Battilus is 1077 nucleotides and 358 residues. The validity of the cDNA-derived amino acid sequence was supported by chemical sequencing of internal tryptic peptides. The molecular masses were calculated to be 39,057 and 39,795 Da, respectively. The amino acid sequence of Liolophura arginine kinase showed 65-68% identity with those of Battilus and Nordotis (abalone) arginine kinases, and the homology between Battilus and Nordotis was 79%. Molluscan arginine kinases also show lower, but significant homology (38-43%) with rabbit creatine kinase. The sequences of arginine kinases could be used as a molecular clock to elucidate the phylogeny of Mollusca, one of the most diverse animal phyla. PMID- 9217007 TI - TGFbeta2 knockout mice have multiple developmental defects that are non overlapping with other TGFbeta knockout phenotypes. AB - The growth and differentiation factor transforming growth factor-beta2 (TGFbeta2) is thought to play important roles in multiple developmental processes. Targeted disruption of the TGFbeta2 gene was undertaken to determine its essential role in vivo. TGFbeta2-null mice exhibit perinatal mortality and a wide range of developmental defects for a single gene disruption. These include cardiac, lung, craniofacial, limb, spinal column, eye, inner ear and urogenital defects. The developmental processes most commonly involved in the affected tissues include epithelial-mesenchymal interactions, cell growth, extracellular matrix production and tissue remodeling. In addition, many affected tissues have neural crest derived components and simulate neural crest deficiencies. There is no phenotypic overlap with TGFbeta1- and TGFbeta3-null mice indicating numerous non-compensated functions between the TGFbeta isoforms. PMID- 9217009 TI - Detection and identification of NAD-catabolizing activities in rat tissue homogenates. AB - NAD may be degraded in several ways. A large number of investigations have shown that at least those catabolic routes which involve the formation of ADP-ribose are related to regulatory processes. In this study a rapid assay was utilized that permits identification of NAD-degrading enzymes directly in sodium dodecylsulfate polyacrylamide gels. Enzymatic activities were recovered by washing the gels in the presence of mild detergents such as lauryl dimethylamine N-oxide or Triton X-100. Subsequent incubation of the gels in the presence of the fluorescent analog 1,N6 etheno-NAD visualized NAD-degrading enzymes. Following excision of the fluorescent bands from the gels, the actual activity of the proteins was established by incubating the gel slices with 14C-labeled NAD and subsequent product analysis by thin layer chromatography (TLC). Homogenates from rat renal cortex and spleen were analyzed by this procedure. While in the spleen homogenate only a single band could be 'activity-stained', in the kidney three bands were detected. Kidney proteins with apparent molecular masses of about 210,000 and 105,000 Da were identified as phosphodiesterase and NAD pyrophosphatase (alkaline phosphodiesterase I), respectively. The third protein exhibited an apparent molecular mass of 41,000. The spleen protein (apparent molecular mass 45,000 Da) cleaved NAD to nicotinamide and ADP-ribose identifying it as NAD glycohydrolase. The procedure is suitable to screen for NAD-converting activities in crude extracts. It is specific for proteins which function as monomers or homo-oligomers. PMID- 9217010 TI - COS-1 cell expression and one-step affinity protein purification and activity of epitope-tagged human erythropoietin and of site-directed mutants. AB - Recombinant human erythropoietin (rhEPO) is an important glycoprotein hormone which has been successfully used in the treatment of anaemia. To facilitate the rapid evaluation of wild-type and mutant forms of rhEPO in structure-function studies, we have developed an expression system in which the recombinant hormone is tagged at the C-terminus with a c-myc peptide. One-step affinity purification of culture supernatants on an anti-myc antibody column yielded proteins which were greater than 50% pure with a specific activity of 300,000 U/mg, in agreement with the value of wild-type protein. We conclude that the additional myc-peptide does not affect receptor binding. The expression system was used to study three mutants in which the N-glycosylation sites were changed to cysteines (Asn24Cys, Asn38Cys and Asn83Cys). Specific activities of these cysteine mutants were significant, but reduced (60%, 22% and 70%, respectively), compared to wild-type. The reduction in specific activity may be due to reduced stability of the mutant proteins. PMID- 9217011 TI - Characterization of the heparin-binding domain of human extracellular superoxide dismutase. AB - The C-terminal, heparin-binding domain of human extracellular superoxide dismutase (hEC-SOD) has been studied as a fusion to human carbonic anhydrase II (HCAII). This technique allows the properties of the EC-SOD domain to be characterized. At the same time, it allows us to differentiate the contributions from the domain, from those properties originating from other parts of EC-SOD. The fusion of the 27 C-terminal amino acids of hEC-SOD to the C-terminal of HCAII (FusCC) resulted in the formation of a monomeric protein, which binds to heparin Sepharose with approximately the same affinity as the tetrameric hEC-SOD. The structure of the fused C-terminal was characterized by CD and NMR spectroscopy and the data were compatible with the presence of alpha-helical structures as suggested by secondary structure predictions. The NMR data show that the C terminal of FusCC moves independently from the rest of the protein and that its central part is involved in conformational exchange. The NOESY spectra demonstrate that the C-terminal in both FusCC and hEC-SOD binds to heparin, and that arginine side chains take part in the binding. PMID- 9217012 TI - Isolation, partial characterization and localization of a dihydrolipoamide dehydrogenase from the cyanobacterium Synechocystis PCC 6803. AB - A dihydrolipoamide dehydrogenase (LPD; dihydrolipoamide:NAD oxidoreductase, EC 1.8.1.4.) activity has been detected in the cyanobacterium Synechocystis PCC 6803. The enzyme was isolated from the membraneous fraction after detergent solubilization and shown to be homogenous on the basis of SDS-PAGE and N-terminal sequencing. The isolated enzyme had a specific activity of 75 U (mg protein)(-1) and was shown to be a homodimer with an apparent molecular mass of 104 kDa for the dimer and 55 kDa for the subunits. The enzyme contains 1.75 mol noncovalently bound FAD (mol enzyme)(-1) suggesting that each subunit contains 1 mol FAD and that the FAD is fairly tightly associated with the enzyme. N-terminal sequencing gave a contiguous amino acid sequence of 17 residues and showed that the N terminus of the LPD from Synechocystis PCC 6803 has significant homologies to other LPDs sequenced so far. Immunoblot experiments indicated that the enzyme is mainly present in the membrane fraction, and immunocytochemical investigations gave evidence that the LPD in Synechocystis PCC 6803 is located in the periplasma space between the cytoplasma membrane and the peptidoglycan layer. This is the first report on an extracellular, membrane-bound LPD in a cyanobacterium. PMID- 9217014 TI - Conformational isomerism of IgG antibodies. AB - The purpose of this study was to determine why apparently homogeneous IgG antibodies were, in some cases, fractionated into at least two components by liquid-liquid partition chromatography (LLPC) in an aqueous two-phase system. Four mouse monoclonal IgG antibodies, two against albumin, one against IgG and one against thyroxine, were shown to adopt different conformational isomeric forms. The four antibodies existed in an equilibrium between two or three conformational forms, the proportion of which could also be estimated by LLPC. Since LLPC detects mainly conformational differences within the antigen-binding sites of IgG antibodies, it could be concluded that the conformational forms differed with respect to their combining sites. Moreover, the isomeric forms of an antibody directed against a protein antigen, formed antigen-antibody complexes with almost identical surface properties. In contrast, complexes with different surface properties were formed when the hapten or hapten conjugated to BSA was bound. Thus, both the conformational isomers could bind antigen, at least when the antigen was a small hapten or a hapten conjugated to a carrier protein. Our results suggest that six out of 57 monoclonal IgG antibodies exist in equilibrium between at least two conformational forms and the biological significance of this isomerism is discussed. PMID- 9217013 TI - Formation of a ternary complex between GM2 activator protein, GM2 ganglioside and hexosaminidase A. AB - The GM2 activator is a 17 kDa protein required for the hydrolysis of GM2 ganglioside by the lysosomal enzyme hexosaminidase A (HexA). The activator behaves as a substrate binding protein, solubilizing GM2 ganglioside monomers from micelles (in vitro) or membranes (in vivo). However, the activator also shows a high order of specificity for activation of lysosomal hydrolases and has been predicted to form a ternary complex with the heterodimeric enzyme (alphabeta) Hex A and GM2 ganglioside. We demonstrated a transient interaction between HexA and the GM2 activator. A chimeric protein containing the FLAG epitope sequence upstream of the GM2 activator was expressed in Escherichia coli and purified using the M1 immunoaffinity (anti-FLAG) column. Binding of the FLAG GM2 activator (FLAG-AP) fusion protein to the M1 column led to the specific retardation of Hex A applied to the column. Other proteins were not retarded by the column nor did they compete with Hex A for binding to FLAG-AP. Hex A and GM2 ganglioside could be simultaneously bound to the column, but the binding of each ligand was independent of the other. The homodimeric (beta beta) isozyme Hex B did not bind to the immobilized activator. The alpha alpha homodimer, HexS, bound weakly, confirming that a hexosaminidase alpha subunit is required for interaction of enzyme and activator. PMID- 9217015 TI - Kinetic studies on the binding of 1,N6-etheno-NAD+ to glutamate dehydrogenase from Clostridium symbiosum. AB - The mechanism of the binding of reduced coenzyme (NAD+) to clostridial glutamate dehydrogenase (GDH) was determined by transient kinetics. The fluorescent 1,N6 ethenoadenine analogue of NAD+ (epsilonNAD+) was used as a probe of nucleotide binary and ternary complex formation because the binding of NAD+ is optically silent. The kinetics of epsilonNAD+ binding were consistent with a 3-step binding process. The enzyme was found to oscillate between two conformational forms, termed E1 and E2, in the presence and absence of L-glutamate. However, L glutamate shifted the equilibrium from 96.8% to 99% of the enzyme in the E1 form. The rapid-equilibrium binding of epsilonNAD+ to the E2 form was rate limited by a slow isomerisation of the ternary complex as the binary complex became saturated with epsilonNAD+. The L-glutamate binary complex had a greater affinity for the coenzyme (Kd = 11 microM) than the free enzyme (Km = 39 microM), indicative of a positive interaction of the substrate and coenzyme binding sites. Steady-state studies were also indicative of a positive interaction in the formation of the catalytic complex, with this complex having a Kd for epsilonNAD+ of 6.8 microM. Consequently, there is stabilization of successive complexes on the reaction pathway. PMID- 9217016 TI - Prediction of secondary structures of proteins using the sequence and spectroscopical data. AB - An algorithm is presented which modifies the parameter of given methods of prediction of secondary protein structures by comparing the predictions with the frequency of secondary structures derived from infrared spectra in a way that the predictions align to the given data. Depending on the prediction method and accuracy of the given secondary structures a 1-6% increase in accuracy can be reached. The algorithm compares the difference between the predicted and real frequency of a given secondary structure in the protein and modifies accordingly the parameter used in the prediction method in order to give a new, more accurate prediction. A correlation between the accuracy of the prediction and increasing correctness between the prediction and infrared data was found using a set of 39 proteins. PMID- 9217017 TI - Characterization of a natural larger form of the antifungal protein (AFP) from Aspergillus giganteus. AB - Two major proteins, alpha-sarcin and an antifungal polypeptide (AFP), are secreted by the mould Aspergillus giganteus MDH 18894 when it is cultured for 70 80 h. A third major protein is also found in the extracellular medium at 48-60 h, but it disappears as the culture proceeds. This protein has been isolated and characterized in terms of apparent molecular mass, electrophoretic and chromatographic behaviour, NH2-terminal primary structure, amino acid content, spectroscopical features, reactivity against anti-AFP antibodies, and antifungal activity. Based on the obtained results it would be an extracellular inactive precursor form of AFP, designated as the large form of AFP (lf-AFP). Its amino acid composition is identical to that of AFP but containing six extra residues. NH2-terminal sequence analysis of the first eight amino acid residues of this polypeptide revealed that the extra residues can be perfectly accommodated within the DNA-deduced sequence of the precursor form of AFP. Its alignment with precursor sequences of different proteins, secreted by a variety of Aspergillus spp., reveals the existence of a common tetrapeptide at the carboxy-terminal end of their leader peptides. This sequence would be Ile/Leu-Xaa-Yaa-Arg, being mostly Xaa and Yaa an acid residue (Asp/Glu) and alanine, respectively. The presence of lf-AFP as an extracellular protein would be in perfect agreement with the existence of this tetrapeptide motif, that can be involved in the protein secretion mechanisms of filamentous fungi. PMID- 9217018 TI - Effect of site-directed mutagenesis of His373 of yeast enolase on some of its physical and enzymatic properties. AB - The X-ray structure of yeast enolase shows His373 interacting with a water molecule also held by residues Glu168 and Glu211. The water molecule is suggested to participate in the catalytic mechanism (Lebioda, L. and Stec, B. (1991) Biochemistry 30, 2817-2822). Replacement of His373 with asparagine (H373N enolase) or phenylalanine (H373F enolase) reduces enzymatic activity to ca. 10% and 0.0003% of the native enzyme activity, respectively. H373N enolase exhibits a reduced Km for the substrate, 2-phosphoglycerate, and produces the same absorbance changes in the chromophoric substrate analogues TSP1 and AEP1, relative to native enolase. H373F enolase binds AEP less strongly, producing a smaller absorbance change than native enolase, and reacts very little with TSP. H373F enolase dissociates to monomers in the absence of substrate; H373N enolase subunit dissociation is less than H373F enolase but more than native enolase. Substrate and Mg2+ increase subunit association in both mutants. Differential scanning calorimetric experiments indicate that the interaction with substrate that stabilizes enolase to thermal denaturation involves His373. We suggest that the function of His373 in the enolase reaction may involve hydrogen bonding rather than acid/base catalysis, through interaction with the Glu168/Glu211/H2O system, which produces removal or addition of hydroxyl at carbon-3 of the substrate. PMID- 9217019 TI - Isolated Bacillus subtilis HemY has coproporphyrinogen III to coproporphyrin III oxidase activity. AB - Oxidation of coproporphyrinogen III to coproporphyrin III is found in extracts of Escherichia coli cells containing the Bacillus subtilis HemY protein (M. Hansson and L. Hederstedt, J. Bacteriol. 176, 5962-5970). We have analysed whether this activity is due to the heterologous expression system, since it in vivo would lead to disruption of the heme biosynthetic pathway. B. subtilis hemY was fused in its 3'-end to a polynucleotide encoding six histidine residues and expressed from plasmids in both E. coli and B. subtilis. The His6-tagged HemY protein extracted from membranes using non-ionic detergent was purified by Ni2+ affinity chromatography. Isolated HemY fusion protein synthesised in E. coli and B. subtilis oxidised coproporphyrinogen III to coproporphyrin III. No direct formation of protoporphyrin IX from coproporphyrinogen III could be detected. Our results suggest that the coproporphyrinogen III to coproporphyrin III activity of HemY is either avoided in B. subtilis in vivo or that coproporphyrin III is a heme biosynthetic intermediate in this bacterium. PMID- 9217020 TI - The possible role of myosin A1 light chain in the weakening of actin-myosin interaction. AB - The effects resulting from the removal of the N-terminus of myosin A1 by limited papain cleavage are investigated. The myosin and heavy meromyosin K+-ATPase and Ca2+-ATPase activities, and actin-activated ATPase activity of heavy meromyosin (HMM) and subfragment-1, are studied. Myosin and HMM preparations devoid of the A1 N-terminus exhibits lower Ca2+-ATPase activities at low ionic strength whereas no differences in K+- or Ca2+-ATPase activities are observed at high ionic strength. Direct binding of actin to monomeric myosin under K+-activated ATPase conditions is much more effective for myosin containing a shortened A1 light chain. The kinetic constants K(app) for actin and V(max) are calculated from actin-activation curves for HMM and subfragment-1. The kinetic constants for HMM are determined under conditions assuring saturation of regulatory light chains (RLC) either with Mg2+ or Ca2+. The removal of the A1 N-terminus influences the actin-myosin interaction in a Ca2+- and phosphorylation-dependent manner; in most cases, this leads to an increase in affinity. In the case of subfragment-1, the removal of the N-terminus of A1 led to a decrease in affinity. It is reasonable to assume that the intact A1 light chain may cause weakening of the actin-myosin interaction under certain conditions. This weakening may be regulated by RLC phosphorylation and RLC-bound calcium-for-magnesium exchange. Such an effect requires a structural minimum that is present in HMM but not in subfragment-1. Implications of such a role for the A1 N-terminus in the myosin-actin interaction are discussed. PMID- 9217021 TI - Microheterogeneity and intrahepatic localization of human and rat liver cytosolic alanine aminotransferase. AB - Native human cytosolic alanine aminotransferase (EC 2.6.1.2) was found to be a homodimer consisting of two 55 kDa subunits. The human enzyme was more cationic and more susceptible to heat inactivation and heavy metal-inhibition than its rat homologue. Isoelectric focussing separated three human isoforms and four rat isoforms of cALT that differed in their isoelectric points. Two subtypes of the enzyme which differed in apparent molecular weight on sodium dodecylsulfate polyacrylamide gel electrophoresis were detected in rat, the liver type and the muscular type. This microheterogeneity was not found for human cytosolic alanine aminotransferase. Immunohistochemical studies revealed that the rat liver enzyme was exclusively localized in periportal hepatocytes, consistent with its role in gluconeogenesis. In human hepatocytes the plasma membrane was intensely stained, implicating an intracellular localization near the plasma membrane. PMID- 9217022 TI - Inhibition of human serine proteases by SPAAT, the C-terminal 44-residue peptide from alpha1-antitrypsin. AB - SPAAT has previously been shown to be a competitive inhibitor of the model serine protease, chymotrypsin. We now present evidence that SPAAT is likewise a competitive inhibitor of human neutrophil elastase and cathepsin G with Ki's of 15-20 and 40 microM, respectively. The mechanism of this inhibition was investigated by comparing the relative effectiveness of the 23-residue N-terminal fragment of SPAAT (N-SPAAT) to inhibit chymotrypsin and human neutrophil elastase. N-SPAAT, which does not contain the primary chymotrypsin cleavage site, was approximately 10-fold less effective as an inhibitor of chymotrypsin than SPAAT (Ki of 65 microM versus 7.5 microM). In contrast, this fragment, which contains the primary human neutrophil elastase cleavage site, was found to competitively inhibit human neutrophil elastase with a Ki of 24 microM which was comparable to that of SPAAT (Ki = 15-20 microM). Thus it appears that SPAAT is a reversible inhibitor of these enzymes rather than an irreversible, stoichiometric one like its parent protein, AAT. Such fragmentation of AAT, however, might provide a mechanism whereby a cascade of decreasingly potent, but increasingly specific SPAAT-related inhibitory peptides could be generated. These results further substantiate the view that SPAAT may play a role in vivo in the protection of extracellular proteins from inappropriate attack by proteases which are elevated during various pathophysiological conditions. PMID- 9217023 TI - One-step purification of histidine-tagged cytochrome bo3 from Escherichia coli and demonstration that associated quinone is not required for the structural integrity of the oxidase. AB - The cytochrome bo3 ubiquinol oxidase from Escherichia coli is a member of the heme-copper superfamily of proton-pumping respiratory oxidases. An improved preparative protocol was desired that would minimize the potential damage during protein isolation of labile mutants of the oxidase. Variants of the oxidase containing a histidine tag at the carboxy-terminus of either subunit I, II or III were constructed. The constructs with the histidine tag on either subunit I or II successfully allowed the enzyme to be isolated with high purity in one step using Ni2+ affinity chromatography. The enzyme with the histidine tag on subunit II is particularly useful insofar as the enzyme isolated in this manner has little, if any, heterogeneity resulting from the presence of heme O in the low spin heme binding site, i.e., cytochrome oo3 is minimized. The enzyme can be prepared in virtually any quantity very rapidly and is suitable for biophysical characterization. Cytochrome bo3 was prepared in either Triton X-100, sucrose monolaurate, or dodecyl maltoside. The enzyme isolated in the presence of either sucrose monolaurate or dodecyl maltoside contains approximately one equivalent of associated ubiquinone, whereas this is absent when Triton X-100 is used. However, the UV/vis absorbance and steady-state kinetic properties of the enzyme are virtually identical regardless of which detergent is used. These data are consistent with previous reports that cytochrome bo3 contains an equivalent of 'tightly associated' ubiquinone, but clearly demonstrate that this quinone can be removed without damaging the enzyme and is not critical to the maintenance of the native structure of the oxidase. PMID- 9217024 TI - Synthetic peptides derived from the central loop of fasciculin: structural analysis and evaluation as inhibitors of acetylcholinesterase. AB - Fasciculins are peptides present in the venom of green and black mamba snakes, with potent inhibitory activity towards acetylcholinesterase. In order to determine the role of fasciculin loop II in the acetylcholinesterase inhibition, two fasciculin fragments were synthesized by the solid phase procedure using N alpha-Boc protected amino acids. The two peptides, Fas-A and Fas-B, span the 26 32 and 22-35 sequences of fasciculin and a disulfide bridge links each peptide end, thus ensuring the formation of a looped structure. Both peptides were characterized chemically, structurally and functionally. Circular dichroism indicated the existence of 19.4 and 24.9% of beta-sheet for Fas-A and Fas-B, respectively; SDS-PAGE patterns and mass spectrometry disclosed the intramolecular disulfide formation in both peptides. An inhibitory effect on eel acetylcholinesterase was observed with the longer peptide (Ki = 15.1 microM), without reaching the affinity level of the parent native toxin (Ki = 0.3 nM). This study confirms that fasciculin central loop residues strongly contribute to toxin interaction with acetylcholinesterase. PMID- 9217026 TI - Depth of invasion of colon carcinoma, lymphatic spread, and laparoscopic surgery. PMID- 9217025 TI - Purification and characterization of diamine oxidase from porcine kidney and intestine. AB - Diamine oxidase, the enzyme catalyzing the oxidative deamination of histamine and other diamines, was purified from porcine kidney and porcine intestine. During all purification steps the enzymes from both tissues showed identical binding and elution characteristics. The native enzymes are homodimeric glycoproteins with an apparent molecular weight of 186 kDa. Under reducing conditions the subunits migrate at 104 kDa on SDS polyacrylamide gels and the deglycosylated subunits migrate at 93 kDa which corresponds to a carbohydrate content of 11%. The native and deglycosylated forms of kidney and intestinal diamine oxidase migrate to the same positions, respectively, on two-dimensional isoelectric focussing/SDS polyacrylamide gels. The sequences of the 21 N-terminal amino acids of both proteins are identical. A polyclonal antibody raised against the kidney enzyme binds equally well to diamine oxidase from both kidney and intestine, inhibits the enzymatic activity, and precipitates all diamine oxidase activity from tissue homogenates. The kidney and intestinal enzymes have identical substrate specificities, efficiently converting aliphatic diamines, histamine, and spermidine. For both enzymes the Km values for histamine, putrescine, and spermidine are 0.02 mM, 0.35 mM, and 3.3 mM, respectively. Spermine, aliphatic monoamines, and aromatic mono- and diamines are poor substrates. In conclusion, the diamine oxidase proteins from porcine kidney and intestine are very likely identical and constitute the only diamine oxidase activity present in these tissues. The structural identity implies identical functions of the proteins in these organs, namely the protection of the organism against high concentrations of diamines. PMID- 9217027 TI - An immunohistochemical assessment of cathepsin D in gastric carcinoma: its impact on clinical prognosis. AB - BACKGROUND: In the context of tumor-associated proteolysis, the prognostic value of cathepsin D in breast carcinoma has been studied but its role is controversial in relation to gastrointestinal carcinoma. The aim of the current study was to determine whether cathepsin D is a prognostic parameter for gastric carcinoma, and also to consider interaction with the urokinase-plasminogen activator (uPA) system as an established risk factor for tumor-associated proteolysis. METHODS: In a consecutive prospective series of 203 gastric carcinoma patients, expression of cathepsin D in tumor cells was semiquantitatively analyzed with immunohistochemistry (scored 0-3). Median follow-up time was 31 months (range, 9 56 months). Kaplan-Meier (log rank) and multivariate Cox analyses were used to analyze survival. RESULTS: Kaplan-Meier analysis (log rank statistics) revealed significant association of increasing cathepsin D detection with poorer disease free survival (P = 0.0042) and poorer overall survival (P = 0.0018) of curatively resected patients. Overall survival of all patients was not significantly correlated. Multivariate analysis of established risk factors for gastric carcinoma, including the uPA system, identified cathepsin D as a new and independent prognostic parameter for disease free survival (P = 0.020; relative risk, 2.98; 95% confidence interval, 1.28-6.91). Plasminogen activator inhibitor type-1 as a representative of the uPA system was confirmed as a strong independent factor for disease free and overall survival. Chi-square analysis showed significant correlation of higher cathepsin D levels with Lauren's diffuse type carcinomas and strong evidence of uPA receptor in tumor cells. However, a subgroup analysis performed according to Lauren's classification revealed a univariate prognostic impact of cathepsin D on both diffuse and intestinal types without independent value. For patients with high levels of uPA receptor (scores of 2 and 3, n = 132), a highly significant association of increasing evidence of cathepsin D with disease free survival (P < 0.0001) and overall survival (P < 0.0001) was observed for curatively resected patients. Significant association with survival was also observed for all patients (P = 0.0407). CONCLUSIONS: Cathepsin D is a new functional prognostic parameter for gastric carcinoma patients with independent value for disease free survival. Moreover, this study indicates that consideration of more than one tumor-associated protease could lead to a more individualized estimation of risk for carcinoma patients. PMID- 9217028 TI - The extent of lymph node dissection for colon carcinoma: the potential impact on laparoscopic surgery. AB - BACKGROUND: The surgeon is no longer able to palpate the mesocolon for lymph node metastases during laparoscopic colectomy. The extent of lymph node dissection should be determined beforehand for cancer control. METHODS: The distribution of lymph node metastases was obtained by the clearing method on colon carcinomas for 164 patients. RESULTS: For pericolic spread: for pT1 tumors, the distance from the primary tumor to a metastatic lymph node was 2.5 cm; for pT2, the distance was within 5 cm; for 97.0 % of pT3 tumors with lymph node metastases, the distance was within 7 cm; for 93.3 % of pT4 tumors with lymph node metastases, the distance was within 7 cm. For central spread: for pT1 tumors, the rate of metastasis to central lymph nodes was 0 %; for pT2, the rate of metastasis was 20.0 % to intermediate lymph nodes; for pT3, the rate of metastasis was 30.6 % to intermediate lymph nodes and 15.3 % to main lymph nodes; for pT4, the rate of metastasis was 44.4 % to intermediate lymph nodes and 22.2 % to main lymph nodes. CONCLUSIONS: Central lymph node dissection is not required for patients with T1 carcinomas, but proximal and distal 3-cm margins of resection are required. For T2, central lymph node dissection that includes the intermediate lymph node should be performed, as well as 5-cm proximal and distal margins of resection. For T3 and T4, central lymph node dissection including the main lymph node should be performed, as well as 7-cm proximal and distal margins of resection. [See editorial on pages 177-8, this issue.] PMID- 9217029 TI - The relation of age, race, and gender to the subsite location of colorectal carcinoma. AB - BACKGROUND: Characteristics that determine the anatomic site within the colorectum where carcinoma is most likely to occur would be useful in choosing optimal modalities for cancer screening and in the study of etiology. The aim of this study was to identify any association of gender, race, or age with colorectal carcinoma in the proximal or distal colorectum. METHODS: A descriptive, population-based epidemiologic study was conducted on colorectal carcinoma (CRC) cases identified through the Illinois Tumor Registry. Among CRC patients diagnosed between 1986 and 1991, age at diagnosis (40-90 years), gender, white or African American race, and subsite of CRC location were identified. Incidence rates were determined with demographic data from the 1990 Illinois census. Logistic regression was used to identify significant subsite specific risk factors for CRC. RESULTS: During the study period, 38,931 cases of CRC occurred in Illinois among whites and African Americans between the ages of 40 and 90 years. Age, race, and gender were independently significantly associated with both proximal and distal CRC risk (P < 0.0001). The greatest risk for white males was for distal CRC, followed by the risk for African American males of developing CRC in the distal colorectum. African Americans were more likely than whites to develop proximal CRC (odds ratio [OR] = 1.19), whereas whites were more likely to have distal CRC than African Americans (OR = 1.26). Regarding the development of CRC anywhere in the colorectum, male incidence rates adjusted for age and race were greater than rates for females (OR = 1.32 for proximal and 1.68 for distal). CONCLUSIONS: The division of the colorectum anatomically in these analyses at the junction of the descending and sigmoid colon, and including the rectum above the anal canal with "distal" CRC, demonstrated a predominance of African Americans among those at risk for proximal colon carcinoma and a predominance of white males among those at risk for distal CRC; these predominances were clearer than in previous reports. These findings may have a role in the selection of optimal CRC screening strategies for each gender or racial group. PMID- 9217030 TI - Thallium-201 single photon emission computed tomography as an indicator of prognosis for patients with lung carcinoma. AB - BACKGROUND: The uptake of thallium-201 (T1-201) in malignant tumors is associated with malignant potential (metastatic potential and proliferative activity). The grade of accumulation of T1-201 in malignant tumors may provide information regarding prognosis. METHODS: A T1-201 single photon emission computed tomography was conducted 120 minutes after intravenous injection of 111 megabecquerel of T1 201 chloride. The authors calculated the uptake ratio to evaluate the degree of T1-201 uptake in the primary tumor. This ratio was compared with survival time and other prognostic factors. RESULTS: The authors studied 152 patients (125 men and 27 women). The group of patients with the low T1-201 uptake ratio survived longer than the group of patients with the high T1-201 uptake ratio (median survival, 58 weeks vs. 33 weeks; P = 0.0138 by the log rank test). The multivariate analysis confirmed that the T1-201 uptake ratio was an independent prognostic factor for survival. CONCLUSIONS: These results suggest that the T1 201 uptake ratio provides independent and objective prognostic information for patients with lung carcinoma. PMID- 9217031 TI - Tumor specific Epstein-Barr virus infection is not associated with leiomyosarcoma in human immunodeficiency virus negative individuals. AB - BACKGROUND: Recent studies have suggested that the Epstein-Barr virus (EBV) is associated with leiomyosarcoma in children with human immunodeficiency virus (HIV) and in organ transplant recipients. To determine whether EBV is associated with leiomyosarcoma in HIV negative patients, the authors examined resected leiomyosarcomas for EBV and HIV. METHODS: Twenty-four leiomyosarcomas were studied and their diagnosis confirmed on pathologic review. From these specimens DNA was isolated. Tumor samples were analyzed for EBV and HIV using a polymerase chain reaction (PCR) technique followed by gel electrophoresis and Southern blot analysis. DNA from an EBV-infected human Burkitt's lymphoma cell line and peripheral blood from an HIV positive patient were used as positive controls for the presence of EBV and HIV, respectively. Immunohistochemistry was performed using an antibody to Epstein-Barr nuclear antigen. RESULTS: HIV was not present in any of the patients analyzed. EBV DNA was detected in tumor tissue; however, 80 cycles of PCR were used before EBV sequences were detected. Therefore, the data indicate that tumor tissue was not infected with EBV. The positive results observed after 80 cycles of PCR were likely due to infiltrating lymphocytes. Immunohistochemistry confirmed the lack of active or latent EBV infection in tumor cells. CONCLUSIONS: The results indicate that EBV is not associated with sporadic leiomyosarcoma in HIV negative patients. Therefore, the biology of leiomyosarcoma associated with HIV may be substantially different from the more common sporadic form. PMID- 9217032 TI - Molecular determination of the malignant potential of smooth muscle neoplasms. AB - BACKGROUND: The determination of the malignant potential of smooth muscle neoplasms remains ambiguous, and yet has far reaching clinical, therapeutic, and social implications. METHODS: In this pilot study, the authors examined smooth muscle isoactin gene expression by polymerase chain reaction in a variety of smooth muscle tumors. RESULTS: A lack of gamma-smooth muscle isoactin gene expression correlated 100% with a pathologic diagnosis of sarcoma. These results suggest that gamma-smooth muscle isoactin gene expression represents a unique molecular marker of oncogenic transformation. CONCLUSIONS: gamma-Smooth muscle isoactin gene expression provides a valuable molecular adjunct to the diagnosis of smooth muscle neoplasms. PMID- 9217034 TI - Effects of medical and psychotherapeutic treatment on the survival of women with metastatic breast carcinoma. AB - BACKGROUND: The authors previously reported a statistically significant effect of psychosocial intervention on survival time of women with metastatic breast carcinoma. In this study, the authors investigated whether this effect could be explained by differences in the medical treatment patients received subsequent to their group participation or differences in causes of death. METHODS: Of the original 86 study participants, medical treatment charts for 61 and death certificates for 83 were available for further analysis. The authors reviewed the course of the medical treatment they received subsequent to their entry into the randomized psychotherapy trial. RESULTS: Although there were no statistically significant differences with regard to chemotherapy and hormone therapy between the control and treatment groups, women in the control group tended to have received more adrenalectomies, although this procedure did not account for the difference in survival time between the control group and the treatment group. Furthermore, women in the control group developed more bone and lung metastases than the women in the treatment group. CONCLUSIONS: Differences in disease course between the control and treatment groups appeared to be independent of any differences in medical treatment received. PMID- 9217033 TI - A randomized phase II trial of two dosage levels of letrozole as third-line hormonal therapy for women with metastatic breast carcinoma. AB - BACKGROUND: It is common practice to utilize a series of different hormonal agents in the treatment of postmenopausal women who, despite disease progression, continue to be candidates for hormonal therapy on a clinical basis. Letrozole is a new highly selective and potent aromatase inhibitor. There are limited data on third-line hormonal therapy in general, and this study was undertaken to evaluate letrozole in this context. METHODS: A randomized trial involving two independent Phase II trials of two letrozole dosage levels, 0.5 mg and 2.5 mg per day, was performed. Eligibility requirements included failure on two prior hormonal therapies and measurable or evaluable disease. RESULTS: Ninety-one patients, 46 receiving 0.5 mg and 45 receiving 2.5 mg of letrozole per day, were assessable for response. At the lower dose, 9 patients (20%) achieved an objective response; 6 patients (13%) had this documented on 2 occasions separated by 3 months. At the higher dose, 10 patients (22%) achieved a response; 8 patients (18%) had this documented on 2 occasions separated by 3 months. The median times to progression were 97 days for the lower dose and 154 days for the higher dose. Toxicity was considered acceptable. CONCLUSIONS: Letrozole has definite antitumor activity as third-line hormonal therapy for women with metastatic breast carcinoma at doses of 0.5 and 2.5 mg per day. It is an effective and generally well-tolerated hormonal agent. PMID- 9217035 TI - Racial and urban/rural differences in cervical carcinoma in Georgia Medicaid recipients. AB - BACKGROUND: The authors conducted a study of racial and geographic differences in the occurrence of cervical carcinoma in a population of uniformly low economic status: Georgia Medicaid recipients. METHODS: Medicaid reimbursement claims data for 1992 were used to calculate counts, rates, and black-to-white risk ratios for newly and previously diagnosed cases of cervical carcinoma in metropolitan Atlanta and in the remainder of the state. RESULTS: Among 615,787 female Georgia Medicaid recipients in 1992, 2050 women (333 per 100,000) had a diagnosis of carcinoma of the cervix. Of 111,208 women who had received Medicaid assistance continuously from 1988 to 1992 (5-year eligibles), a new claim for cervical carcinoma was submitted for 110 (99 per 100,000). In both analyses, rates were higher in metropolitan Atlanta than in the remainder of the state. Black women had significantly higher claims rates than white women only in metropolitan Atlanta; risk ratios were 3.7 (95% confidence interval [CI], 1.3-10.8) for new claims among 5-year eligibles, and 3.5 (95% CI, 3.0-4.1) for prevalence. There was no racial disparity in cervical carcinoma rates in rural areas. CONCLUSIONS: The current study data suggest a high risk of cervical carcinoma among metropolitan Atlanta Medicaid recipients, particularly blacks. Data from rural Georgia (but not Atlanta) support the hypothesis that racial differences in cervical carcinoma rates would largely disappear in a population of uniform economic status. PMID- 9217036 TI - Effects of endocrine therapy on the primary lesion in patients with prostate carcinoma as evaluated by endorectal magnetic resonance imaging. AB - BACKGROUND: Little effort has been made at the quantitative and qualitative evaluation of patients with prostate carcinoma, including downsizing and downstaging of the primary lesion, after conservative therapy. The current study was undertaken to investigate the qualitative and quantitative effects of endocrine therapy on the primary prostate carcinoma using magnetic resonance imaging (MRI). METHODS: The primary prostate carcinoma was evaluated by endorectal MRI approximately 4 months after the initiation of endocrine therapy in 48 patients with histologically confirmed prostate carcinoma detected by endorectal MRI before therapy. RESULTS: The volumes of the prostate gland, the carcinoma, and the noncarcinomatous components were reduced to 60.2 +/- 2.7%, 25.5 +/- 2.9%, and 83.2 +/- 6.3% of their pretreatment volumes respectively after endocrine therapy, indicating that the tumors are more susceptible to endocrine therapy than the nontumorous components. The number of prostate carcinomas that demonstrated low signal intensity compared with the normal peripheral zone on T2 weighted images decreased after endocrine therapy and the number of carcinomas with enhancement of T1-weighted contrast-enhanced images increased after therapy. Seven of the 48 patients underwent downstaging after endocrine therapy, based on the endorectal MRI evaluation. CONCLUSIONS: The results of the current study suggest that downsizing and occasionally downstaging of the carcinoma may occur after endocrine therapy in patients with prostate carcinoma. In addition, the androgen sensitivity of the prostate carcinoma tissue is relatively high compared with the residual noncancerous prostate gland. PMID- 9217037 TI - Retinoblastoma protein expression and MIB-1 correlate with survival of patients with malignant astrocytoma. AB - BACKGROUND: Regulatory coupling of cell proliferation and apoptosis is suggested by recent findings with regard to certain tumors, such as the finding of tumor resistance to anticancer therapy caused by inhibition of apoptosis. The processes leading to apoptosis appear to be regulated by a variety of oncogenes and tumor suppressor genes. In the current study, the relation between apoptosis and expression of retinoblastoma protein (pRB) was assessed in 50 primary anaplastic astrocytomas (AAs) and 46 recurrent tumors in the same patients as the primary tumors after macroscopic total surgical resection and chemoradiotherapy. METHODS: Apoptotic cells were identified by the in situ 3'-end labeling technique. Proliferative potential, pRB expression, and p53 expression were evaluated immunohistochemically using anti-Ki-67 (MIB-1), anti-pRB, and anti-p53 antibodies, respectively. The prognostic value of these biologic markers in AA patients undergoing treatment was also evaluated. RESULTS: The mean apoptotic index (AI) was 0.91 +/- 0.70% for specimens obtained at the initial surgery and 2.32 +/- 1.71% for those obtained at recurrence. There was no apparent correlation between the AI and the MIB-1 staining index (MI) in primary AAs, whereas significantly higher AI and MI were observed in recurrent pRB negative cases than in their pRB positive counterparts. The survival of patients with AAs showing a high MI and negative pRB immunostaining was significantly shorter than in the other cases. Neither the size of the apoptotic fraction nor the p53 expression in primary tumor correlated with the overall survival. CONCLUSIONS: The clinical outcome of patients with AA may be associated with aberrant pRB function and increased proliferative activity rather than an inability of tumor cells to undergo apoptosis. PMID- 9217038 TI - Efficacy of postacute brain injury rehabilitation for patients with primary malignant brain tumors. AB - BACKGROUND: Patients with primary malignant brain tumors (PMBT) often have neurobehavioral deficits due to the tumor, subsequent surgery, and therapies that interfere with their ability to live independently or work. Previous studies have shown that such patients generally have a progressive decline in functioning from diagnosis to death. Consequently, PMBT patients have not been considered good candidates for rehabilitation services. The current study is a preliminary, retrospective investigation of the effectiveness of postacute brain injury rehabilitation methods, originally developed for traumatic brain injury survivors, in a sample of patients with PMBT. METHODS: The subjects were 13 patients with a history of surgical resection of PMBT and subsequent radiation and chemotherapy. There were 8 males and 5 females with a mean age of 34.3 +/- 10.0 years and a mean educational level of 15.1 +/- 1.7 years. Mean time from tumor diagnosis to the commencement of rehabilitation was 75.4 +/- 87.9 months. All patients had cognitive deficits documented with neuropsychologic tests. Patients received an average of 2.6 +/- 1.9 months of postacute brain injury rehabilitation. RESULTS: Six patients had increased independence during the time from the start of rehabilitation to discharge, six were unchanged, and one patient had decreased independence. Eight patients had increased productivity during the same time period, four were unchanged, and one had decreased productivity. Treatment gains were maintained at follow-up 8.0 +/- 7.6 months after discharge. CONCLUSIONS: The results of the current study offer preliminary support for the effectiveness of postacute brain injury rehabilitation in the management of PMBT patients. Although additional investigation is needed, such treatment appears to be an attractive, relatively low cost option for these patients. PMID- 9217039 TI - Self-reported comprehensive health status of adult brain tumor patients using the Health Utilities Index. AB - BACKGROUND: The comprehensive health status of adult survivors of brain tumors is largely unexplored. METHODS: Using a multiattribute approach embodied in a 15 item self-assessment questionnaire, the overall burden of morbidity was measured in 50 brain tumor patients who were attending a neurooncology outpatient clinic. The comprehensive health status was accorded utility scores, and comparisons were made with health status measurements of the general population. RESULTS: The questionnaire was completed with ease by 90% of the respondents. Among the respondents, only 10% of the patients did not report some form of morbidity, and 80% reported multiple impairments. The most prevalent impairments occurred in the attributes of sensation, emotion, and cognition (in this predominantly ambulant group); each of these elements was limited in the majority of patients. A surprising finding was the self-report of pain by nearly 50% of the respondents. CONCLUSIONS: In this group of patients, the burden of morbidity and its complexity greatly exceeded that reported for the general population and were inadequately revealed by Karnofsky performance scores. The use of multiattribute health status measurement tools offers numerous advantages and should be employed in the routine clinical management of cancer patients. PMID- 9217040 TI - Hyperfractionated radiation therapy and 5-fluorouracil, cisplatin, and mitomycin C (+/- granulocyte-colony stimulating factor) in the treatment of patients with locally advanced head and neck carcinoma. AB - BACKGROUND: The authors had previously reported preliminary results of a treatment regimen of concurrent hyperfractionated radiation therapy and chemotherapy in patients with locally advanced head and neck carcinoma that demonstrated both feasibility and high local control. In an attempt to reduce acute mucosal and hematologic toxicity, granulocyte-colony stimulating factor (G CSF) was added during the second phase of this study. METHODS: Seventy patients (53 with Stage IV and 17 with Stage III disease) were entered between May 1988 and June 1995 into a Phase I/II trial of concurrent radiation therapy (74.4 gray (Gy) total dose; 1.20 Gy twice daily), 5-fluorouracil (1000 mg/m2/24 hours for 72 hours), and cisplatin (50 mg/m2) for 3 cycles with the addition of mitomycin C (8 mg/m2) in Cycle 2. G-CSF was added after the initial entry of 34 patients. RESULTS: At a median follow-up of 41 months (range, 12-80 months), 44 patients were alive with a projected median overall survival of 54 months. Grade 3/4 mucositis, observed in 65% of patients, was equally prevalent and prolonged in both G-CSF-treated (+) and G-CSF-naive (-) patients. Grade 3/4 leukopenia was present in 45% and 36% of G-CSF- and G-CSF+ patients, respectively. The 3-year locoregional control and cause specific survival rates were 68% and 75%, respectively. CONCLUSIONS: This regimen was feasible and effective but caused severe mucositis. No benefit was derived from the addition of G-CSF. This regimen deserves further modification to reduce acute mucositis toxicity yet maintain the high locoregional control rate. PMID- 9217041 TI - Signet-ring sinus histiocytosis: a reactive disorder that mimics metastatic adenocarcinoma. AB - BACKGROUND: Signet-ring sinus histiocytosis is a rare and distinctive reactive disorder recently observed in the axillary lymph nodes of patients with breast carcinoma. This form of sinus histiocytosis closely resembles and can easily be confused with metastatic adenocarcinoma. METHODS: To determine the incidence of this reactive process in lymph nodes from different anatomic sites, broaden its morphologic spectrum, and discuss the differential diagnosis, the authors examined lymph nodes from 316 radical prostatectomy specimens, 184 modified radical mastectomy specimens, 108 colectomy specimens, 57 gastrectomy specimens, and 27 radical hysterectomy specimens. These surgical procedures were performed to treat carcinoma of the prostate, breast, colon, stomach, and uterine cervix, respectively. A total of 9741 lymph nodes were histologically examined. The lymph nodes containing sinus signet-ring cells were stained with mucicarmine, Alcian blue, and periodic acid-Schiff stains (PAS). Immunostains for epithelial, lymphoid, and histiocytic markers were also performed. In two cases, tissue was retrieved from the paraffin block and subsequently processed for electron microscopic examination. RESULTS: Only 4 of 316 radical prostatectomy specimens (1.2%) and 2 of 184 axillary dissections (1.08%) contained lymph nodes with signet-ring sinus histiocytosis. Of 9741 lymph nodes reviewed, 37 (24 pelvic and 13 axillary lymph nodes) had signet-ring sinus histiocytosis, for an incidence of 0.38%. Microscopically, the signet-ring histiocytes lacked nuclear atypia and were mucin negative. In two cases, clusters of histiocytes with rounded, eosinophilic, diastase resistant, PAS positive cytoplasmic globules were observed. Both types of signet-ring cells showed reactivity for histiocytic markers and were negative for cytokeratin and lymphoid markers. By electron microscopy, most histiocytes were shown to have a large empty vacuole that displaced the nucleus. Granular material was observed in some of the vacuoles. Some histiocytes exhibited a rounded cytoplasmic body composed of central electron dense, granular material and a rim of microfibrils. No lipid droplets were identified. CONCLUSIONS: Signet-ring sinus histiocytosis is a rare and distinctive reactive disorder found incidentally in the pelvic and axillary lymph nodes of patients with carcinoma of the prostate and breast, respectively. Although this histiocytic reaction mimics metastatic adenocarcinoma and signet ring cell lymphoma, it can be identified by careful cytologic analysis together with positive reactivity for histiocytic markers, negative mucin stains, and lack of reactivity for epithelial and lymphoid markers. The etiology and pathogenesis of this unusual form of sinus histiocytosis remains unclear. PMID- 9217042 TI - Fatal pulmonary toxicity resulting from treatment with gemcitabine. AB - BACKGROUND: Pulmonary toxicity reported with gemcitabine is usually mild and self limiting. The authors report a series of three patients who had life-threatening pulmonary toxicity after receiving gemcitabine. METHODS: The three patients presented to two major teaching hospitals with significant pulmonary dysfunction while receiving gemcitabine. Case data were obtained from patient records. A review of the literature was done to seek reports of pulmonary toxicity with gemcitabine and cytosine arabinoside (ara-C). RESULTS: The common features of the respiratory illnesses of the three patients in this study were tachypnea, marked hypoxemia, and an interstitial infiltrate on chest radiograph consistent with pulmonary edema. There was no evidence of underlying heart disease in any patient. In addition, there was no evidence of infection, metabolic causes, or lymphangitic carcinomatosis to explain the clinical findings. Two patients died, and postmortem examination confirmed acute RDS (respiratory distress syndrome), whereas in the third patient a transbronchial biopsy showed interstitial pneumonitis. These findings were consistent with drug-induced pulmonary toxicity. Diuretics and corticosteroids were useful measures for treating the patients' symptoms, and one patient survived after gemcitabine was withdrawn. CONCLUSIONS: These three cases of acute RDS may be the result of a capillary leak phenomenon due to treatment with gemcitabine, as observed in patients given intermediate dose and high dose ara-C, a drug similar in structure and metabolism to gemcitabine. The authors suggest caution in repeated administration of gemcitabine to patients who develop unexplained noncardiogenic pulmonary edema. Withdrawing gemcitabine and administering corticosteroids and diuretics may help to avert a fatal outcome. PMID- 9217043 TI - Primary gastric T-cell lymphoma with and without human T-lymphotropic virus type 1. AB - BACKGROUND: Gastric T-cell lymphomas are rare, and their incidence and viral status have not yet been fully clarified. METHODS: Sixty-seven cases of surgically resected gastric lymphomas from city hospitals in Tokyo were evaluated. The surface phenotype was determined by immunohistochemistry, gene rearrangement by Southern blot hybridization, association with Epstein-Barr virus (EBV) by EBV-encoded small RNAs in situ hybridization, and the presence of human T-cell lymphotropic virus type 1 (HTLV-1) by serology, Southern blot hybridization, and polymerase chain reaction analysis. RESULTS: Five of the 67 cases were T-cell lymphoma (7%): 3 cases were HTLV-1 negative (-) and 2 were HTLV 1 positive (+). Systemic eosinophilia was observed in the three HTLV-1(-) gastric lymphomas. Neoplastic cells were morphologically similar in both groups, but a granulomatous reaction with marked eosinophilia was observed only in the two cases of HTLV-1(-) lymphoma. They also had characteristics of natural killer (NK) cell-like T-cell lymphoma, expressing NK markers and TCRgamma gene rearrangement. Positivity with HML-1 (specific for intestinal epithelial T-cells lymphoma was observed in one HTLV-1(+) lymphoma. The EBV gene was detected in only one case of B-cell lymphoma but not in any case of T-cell lymphoma. CONCLUSIONS: Gastric T cell lymphoma occurs in 7% of gastric lymphomas in Japan and is comprised of HTLV 1-related lymphomas and lymphomas unrelated to HTLV-1, including NK cell-like lymphomas with eosinophilia. PMID- 9217044 TI - Prenatally diagnosed neuroblastoma. AB - BACKGROUND: Prenatally diagnosed neuroblastomas have been reported in increasing numbers over the past several years, and there are now a few reviews based on up to 21 cases. The purpose of this article is to review the clinical and biologic features of prenatally diagnosed neuroblastoma based on a review of 55 cases. METHODS: A review was conducted of 3 cases seen at the study institution and 52 other cases reported thus far in the literature. RESULTS: Prenatal diagnosis was made usually after 32 weeks of gestation. Approximately 93% of the tumors were adrenal in origin, and 44% of these were cystic. Thirty-seven patients (67%) had Stage I disease, 12 (22%) had Stage IV-S disease, and only 3 (5%) had Stage IV disease. The DNA index was favorable (> 1) in 14 of 16 patients studied. None of these 16 patients studied had amplification of the N-myc oncogene. Catecholamines were elevated in only 33% of the patients. The liver was the most common site of dissemination, which was observed in 25% of patients; bone involvement was not observed in any patient. Ultrasonography failed to detect existing hepatic metastasis in three patients. Primary surgical resection was performed in 47 patients (85%). Chemotherapy was given to five patients and radiotherapy to three. Of the 50 patients for whom follow-up information was available, 45 (90%) were alive at a range of 2-120 months from diagnosis. CONCLUSIONS: Prenatally diagnosed neuroblastomas are predominantly adrenal in origin and frequently cystic. The liver is the most common site of dissemination and bone involvement is notably absent. The vast majority of these infants have a favorable stage of disease (I, II, and IV-S) and favorable biologic features, and consequently have an excellent prognosis. Although surgery alone is curative for most patients, a period of observation may avoid surgery in some individuals who may achieve spontaneous regression. PMID- 9217045 TI - A phase II study of carboplatin as a treatment for children with acute leukemia recurring in bone marrow: a report of the Children's Cancer Group. AB - BACKGROUND: Carboplatin is an analogue of cisplatin with less nonhematologic toxicity and a similar spectrum of antineoplastic activity. Although cisplatin has not been found to be an active agent against leukemia, carboplatin-induced complete remissions have been observed in adults with acute myelogenous leukemia (AML), and antileukemic activity has been observed in a Phase I trial involving children with acute lymphoblastic leukemia (ALL) and AML. Therefore, a pediatric Phase II study was undertaken to determine the degree of activity of carboplatin in childhood ALL and AML. METHODS: Between October 1991 and November 1994, the Children's Cancer Group conducted a Phase II study of carboplatin given by 5-day continuous intravenous infusion to children with acute leukemia recurring in bone marrow. RESULTS: Minimal antileukemic activity was demonstrated in patients with ALL and AML. One of 21 eligible patients with ALL achieved a partial response. Of 23 eligible patients with AML, including 1 patient with chronic myelogenous leukemia in blast crisis, 1 had hypocellular M1 bone marrow with a platelet count of 15,000/mm3, and 2 achieved partial responses. Nonhematologic toxicities, which were infrequent, included mild hepatic and renal dysfunction. CONCLUSIONS: In this pediatric Phase II trial of carboplatin as a treatment for acute leukemia, minimal activity was demonstrated in patients with ALL and AML recurring in bone marrow. Further evaluation of carboplatin as a treatment for childhood leukemia, using the dose schedule of 216 mg/m2/day given by 5-day continuous intravenous infusion, does not appear warranted. PMID- 9217046 TI - A phase I/IB trial of murine monoclonal anti-GD2 antibody 14.G2a plus interleukin 2 in children with refractory neuroblastoma: a report of the Children's Cancer Group. AB - BACKGROUND: The murine monoclonal antibody (MoAb) 14.G2a recognizes GD2, a disialoganglioside expressed in tumors of neuroectodermal origin, and facilitates antibody dependent cellular cytotoxicity (ADCC) in vitro. When given in vivo, interleukin-2 (IL-2) can increase ADCC by enhancing the activity and number of circulating lymphocytes. METHODS: Thirty-three pediatric patients with GD2 positive malignancies, ranging in age from 2 to 17 years (median, 9.9 years), received IL-2 and 14.G2a in this Phase I/IB study of the Children's Cancer Group (CCG) and were monitored for toxicities and response to therapy. Seven of these patients also received granulocyte-macrophage-colony stimulating factor. RESULTS: The maximum tolerated dose (MTD) of 14.G2a with IL-2 was 15 mg/m2/day. The most prevalent Grade 3-4 toxicities were generalized pain (n = 14 [42%]) and fever without documented infection (n = 17 [52%]). IL-2 was thought to be the causative agent in most cases of fever. Toxicities attributed to 14.G2a included pain, allergic or anaphylactic reactions, and rash. Human antimouse antibodies were demonstrated in 9 of 21 evaluated patients. One patient with neuroblastoma had a partial response, and one patient with osteosarcoma had a complete response. Immunocytology demonstrated that the number of neuroblastoma cells in bone marrow decreased in three patients. CONCLUSIONS: The murine MoAb 14.G2a was well tolerated at the MTD and appeared to have some antitumor activity. Further development of this approach will involve additional engineered forms of the antibody as well as testing in the adjuvant and minimal residual disease setting. PMID- 9217047 TI - Noncytotoxic drug therapy in children with unresectable desmoid tumors. AB - BACKGROUND: Antiestrogens and nonsteroidal antiinflammatory drugs have been shown to be effective in adult patients with unresectable or recurrent desmoid tumors. It appears that the growth of these tumors is influenced by estrogen, and that antiestrogen treatment may inhibit further proliferation of tumor cells. Nonsteroidal antiinflammatory drugs are thought to be effective through their interference with prostaglandin metabolism. METHODS: Two children with unresectable desmoid tumors (aggressive fibromatosis) were treated with tamoxifen (1 mg/kg orally, twice daily) and diclofenac (2 mg/kg rectally, twice daily). RESULTS: At last follow-up, tumor regression and growth arrest were maintained for more than 51 months in 1 child with rapidly growing recurrent fibromatosis of the thoracic wall. Another child with an inoperable desmoid tumor of the submandibular region had stable disease since the initiation of treatment. CONCLUSIONS: This is the first report describing this treatment approach in childhood fibromatosis. Combined therapy with endocrine therapy and nonsteroidal antiinflammatory drugs may be a nonaggressive alternative to chemotherapy and radiotherapy in the treatment of children with inoperable desmoid tumors. PMID- 9217048 TI - Adjuvant chemotherapy for the treatment of intracranial ependymoma of childhood. AB - BACKGROUND: Current treatment for childhood intracranial ependymomas with surgery and radiation therapy (RT) yields 5-year survival rates ranging from 50-70% after complete resection to 0-30% after incomplete surgical resection. The role of chemotherapy in the treatment of ependymoma has not been established. In this pilot study, children with newly diagnosed intracranial ependymoma were treated with RT and chemotherapy using agents comparable to those found to be active in the treatment of intracranial ependymoma in infants. METHODS: Nineteen children age 3-14 years (median, 7.5 years) were treated with postoperative RT and chemotherapy. Chemotherapy was comprised of carboplatin, 560 mg/m2, with vincristine, 1.5 mg/m2, weekly for 3 weeks, alternating at 4-week intervals with ifosfamide, 1.8 g/m2, and etoposide, 100 mg/m2, for 5 consecutive days for a total of 4 cycles. RESULTS: The 5-year progression free survival (PFS) estimate was 74%. The extent of surgical resection was not a significant prognostic factor in this study. By contrast, ependymomas located in the posterior fossa were associated with a higher rate of progression (P = 0.036). Toxicity, limited predominantly to myelosuppression, was manageable. CONCLUSIONS: The PFS for children with postoperative residual ependymoma treated with RT and chemotherapy in this study was higher than published survival results for RT alone. These results suggest a role for multialkylator chemotherapy in incompletely resected intracranial ependymoma and provide the rationale for a randomized trial comparing this strategy with conventional postoperative RT. PMID- 9217049 TI - Analysis of six DNA components of the faba bean necrotic yellows virus genome and their structural affinity to related plant virus genomes. AB - Faba bean necrotic yellows virus (FBNYV) has a multicomponent circular ssDNA genome. In addition to a previously described genome component (C1) coding for a replicase-associated protein (Rep), five further components (C2 to C6) have now been identified. Each of the six components is about 1 kb in size, contains one major open reading frame (ORF) in the virion sense with a TATA box and polyadenylation signal, and has a noncoding region containing a highly conserved sequence possibly forming a stem-loop structure. Similar to C1, C2 encodes another putative Rep of 33.1 kDa, which is closely related to the Rep of banana bunchy top virus (BBTV). Based on bacterial expression and immunoblot analysis, the ORF of C5 encodes the capsid protein (CP) with a deduced molecular mass of 19 kDa. The FBNYV CP shares the highest amino acid (aa) identity (56.2%) with that of subterranean clover stunt virus (SCSV). The ORF of C4 potentially codes for a hydrophobic protein which appears to be structurally and functionally similar to the BBTV-C4 and SCSV-C1 proteins. No protein sequence similarities were found in databases for the C3 and C6 ORFs of FBNYV. FBNYV is clearly distinct from any known virus but is taxonomically related to BBTV and SCSV. PMID- 9217051 TI - Ultrastructure of HIV-1 genomic RNA. AB - The HIV-1 RNA genome is a dimer which consists of two identical strands of RNA linked near their 5' ends by a dimer linkage structure (DLS). We have structurally characterized full-length HIV-1 genomic RNA isolated from HIV-1 virions by electron microscopy. As in other retroviruses, the HIV-1 RNA genome contains a central dimer linkage structure and additional loop structures within each monomer subunit. In contrast to the DLS of other retroviruses, the DLS region of HIV-1 contains a loop of 323 +/- 44 nucleotides. The free 5' ends of the two RNA strands were not visualized, suggesting that the 5' end regions are involved in interstrand complementary base pairing. Computer modeling identified a single stable structure that was consistent with the electron microscopy data. In this model, the two RNA strands are linked at their 5' ends by two contact points derived from "kissing-loop" interactions between r-u5 and SL1 stem-loops and their counterparts on the second strand. These interactions may contribute to the formation of stable HIV-1 RNA dimers in vivo. PMID- 9217050 TI - Kinetics of cytokine mRNA expression in the central nervous system following lethal and nonlethal coronavirus-induced acute encephalomyelitis. AB - The potential role(s) of cytokines in the reduction of infectious virus and persistent viral infection in the central nervous system was examined by determining the kinetics of cytokine mRNA expression following infection with the neurotropic JHM strain of mouse hepatitis virus. Mice were infected with an antibody escape variant which produces a nonlethal encephalomyelitis and compared to a clonal virus population which produces a fulminant fatal encephalomyelitis. Infection with both viruses induced the accumulation of mRNAs associated with Th1 and Th2-type cytokines, including IFN-gamma, IL-4, and IL-10. Peak mRNA accumulations were coincident with the clearance of virus and there was no obvious differences between lethally and nonlethally infected mice. TNF-alpha mRNA was induced more rapidly in lethally infected mice compared to mice undergoing a nonfatal encephalomyelitis. Rapid transient increases in the mRNAs encoding IL-12, iNOS, IL-1alpha, IL-1beta, and IL-6 occurred following infection. Nonlethal infections were associated with increased IL-12, IL-1beta, and earlier expression of IL-6, while lethal infections were associated with increased iNOS and IL-1alpha mRNA. These data suggest a rapid but differential response within the central nervous system cells to infection by different JHMV variants. However, neither the accumulation nor kinetics of induction provide evidence to distinguish lethal infections from nonlethal infections leading to a persistent infection. Accumulation of both Th1 and Th2 cytokines in the central nervous system of JHMV-infected mice is consistent with the participation of both cytokines and cell immune effectors during resolution of acute viral-induced encephalomyelitis. PMID- 9217052 TI - Characterization of human endogenous retrovirus type K virus-like particles generated from recombinant baculoviruses. AB - The family of human endogenous retrovirus type K (HERV-K) comprises members with long open reading frames (ORF) for retroviral proteins. The existence of a biologically active provirus with replicative capacities has not yet been demonstrated. To confirm the assumption that HERV-K codes for the previously observed retrovirus-like particles (human teratocarcinoma-derived virus, HTDV) in human teratocarcinoma cells, we have constructed recombinant full-length HERV-K cDNA-based baculoviruses with gag, pro, pol, and env ORFs. Two viral constructs were used for infections of insect cells, one bearing 67 bp of the 5' untranslated region upstream of the 5' splice donor (SD) site and of the retroviral genes, the second omitting the SD sequence. For both recombinant viruses, indirect immunofluorescence and laser scan analyses revealed expression of HERV-K Gag protein. Electron microscopy studies demonstrated efficient production of virus-like particles (VLPs) at the cytoplasmic cell membranes. These VLPs are morphologically identical with the HTDV phenotype. In immunoelectron microscopy of ultrathin frozen sections, anti-HERV-K Gag antibodies specifically reacted with HERV-K VLPs. In Western blots, in addition to the 76-kDa precursor protein, the putative major core protein with an apparent molecular mass of 32 kDa exhibited predominant immunoreactivity with anti-Gag antiserum. In contrast, neither HERV-K Env nor cORF proteins could be detected due to inefficient mRNA splicing. Purified particles from insect cell culture supernatants tested in an ultrasensitive reverse transcriptase assay revealed weak polymerase activity. The data demonstrate that HERV-K codes for retroviral particles of the HTDV phenotype. PMID- 9217053 TI - A role for baculovirus GP41 in budded virus production. AB - Insect cells infected with tsB1074, a temperature-sensitive mutant of Autographa californica nuclear polyhedrosis virus, exhibit a "single-cell-infection" phenotype whereby the infection progresses through the very late phase culminating in occlusion body formation, but neighboring cells do not become infected. Marker rescue mapping and DNA sequencing correlated a single nucleotide substitution within the baculovirus gp41 gene with the temperature-sensitive phenotype of tsB1074. The product of the gp41 gene, GP41, is an O-glycosylated protein found in occluded but not budded virions [M. Whitford and P. Faulkner (1992) J. Virol. 66, 3324-3329]. However, budded virus was not produced in tsB1074-infected cells at the nonpermissive temperature of 33 degrees, indicating an additional role for GP41 in budded virus formation. Electron microscopy revealed that nucleocapsids were produced but retained in the nucleus of tsB1074 infected cells at 33 degrees. Thus, GP41 was required for the egress of nucleocapsids from the nucleus in the pathway of budded virus synthesis. PMID- 9217054 TI - Kinetics of cytokine expression and regulation of host protection following infection with molecularly cloned Venezuelan equine encephalitis virus. AB - In the mouse model, the arbovirus Venezuelan equine encephalitis virus (VEE) replicates in lymphoid tissues prior to either inducing protective immunity (attenuated VEE mutant) or progressing to lethal encephalitis (virulent parent VEE). To investigate the mechanism of the protective response, cytokine gene expression was examined during the course of the primary in vivo immune response to molecularly cloned, virulent VEE and a single-site attenuated VEE mutant, using a quantitative reverse transcriptase-polymerase chain reaction assay. VEE induced cytokine gene expression was 100-fold elevated over that of untreated controls for IFN-gamma and IL-6 and 10-fold increased for IL-12, IL-10, and TNF alpha. There was no qualitative difference in cytokine gene induction comparing mice infected with the attenuated and the virulent VEE; however, there were significant differences in the cytokine gene expression kinetics. In mice infected with the attenuated VEE, elevated cytokine gene expression was delayed 24 hr when compared to mice infected with the virulent parent VEE clone at the same dose. Further, IFN-gamma protein secretion by cells from the draining lymph node mimicked the pattern of IFN-gamma gene induction by cells harvested from the same site. IFN-gamma gene expression was elevated at an earlier time point in mice given virulent V3000 24 hr after attenuated V3032 injection compared to mice infected with virulent V3000 alone. The combined V3000/V3032 infection resulted in host protection. Treatment of mice with IL-12 prior to infection with virulent VEE failed to reduce the severity of infection, while anti-IL-12 antibody did not prevent the early protective effect of attenuated virus. In contrast, administration of anti-IFN-alpha/beta antibody prior to VEE infection worsened virulent VEE disease. These results indicate that the attenuated VEE strain elicits a similar but delayed cytokine response compared to the virulent strain, suggesting that the kinetics of cytokine expression and the particular cytokine produced may influence the development of a host protective response. Furthermore, IFN-alpha/beta, but not IL-12, seems to be a major factor in the induction of early protection against VEE infection and disease. PMID- 9217055 TI - A human IgG1 (b12) specific for the CD4 binding site of HIV-1 neutralizes by inhibiting the virus fusion entry process, but b12 Fab neutralizes by inhibiting a postfusion event. AB - The human b12 IgG1, specific for the CD4 binding site of the gp120 of HIV-1, was prepared by recombinant DNA technology. It had a high neutralization rate constant (-3.5 x 10(5) M(-1) sec(-1)), although this is about 10-fold less than the values for the best poliovirus or influenza A virus MAbs. The recombinant b12 Fab neutralized well, with about one-tenth of the activity of b12 IgG. The mechanisms by which b12 IgG1 and its Fab neutralize HIV-1 IIIB on C8166 cells have been investigated. Neither inhibited attachment of virus to the target cell as judged by FACS, immunofluorescence, and ELISA data. This was controlled using MAb F105, another human IgG1, that did neutralize by inhibiting attachment under our conditions. The interactions of b12 IgG- and Fab-neutralized virions with target cells were compared with those of nonneutralized virus using a number of different techniques (fluorescence dequenching of R18-labeled virions, immunofluorescence of virion gp41 and p24 antigens, and acquisition of resistance to removal of virions from the cell by protease). These and the inhibition of HIV 1-mediated cell-cell fusion all demonstrated that b12 IgG neutralized by inhibiting the primary fusion-uncoating mechanism. However, the interactions of b12 Fab-neutralized and nonneutralized virions with C8166 cells were indistinguishable. Thus b12 Fab did not inhibit fusion uncoating, and by inference inhibited a stage of infection that occurs after the entry of the virion core into the cytoplasm. It is therefore possible that b12 IgG kills HIV-1 twice over, by fusion-inhibition and by inhibiting the postentry event proposed for the Fab. The mechanism of neutralization of b12 Fab and of other MAbs that neutralize in a similar way and why b12 Fab and IgG neutralize by different mechanisms are discussed. PMID- 9217056 TI - Expression of interferon-gamma by a coronavirus defective-interfering RNA vector and its effect on viral replication, spread, and pathogenicity. AB - A defective-interfering (DI) RNA of the murine coronavirus mouse hepatitis virus (MHV) was developed as a vector for expressing interferon-gamma (IFN-gamma). The murine IFN-gamma gene was cloned into the DI vector under the control of an MHV transcriptional promoter and transfected into MHV-infected cells. IFN-gamma was secreted into culture medium as early as 6 hr posttransfection and reached a peak level (up to 180 U/ml) at 12 hr posttransfection. The DI-expressed IFN-gamma (DE IFN-gamma) exhibited an antiviral activity comparable to that of recombinant IFN gamma and was blocked by a neutralizing monoclonal antibody against IFN-gamma. Treatment of macrophages with DE-IFN-gamma selectively induced the expression of the cellular inducible nitric oxide synthase and the IFN-gamma-inducing factor (IGIF) but did not affect the amounts of the MHV receptor mRNA. Antiviral activity was detected only when cells were pretreated with IFN-gamma for 24 hr prior to infection; no inhibition of virus replication was detected when cells were treated with IFN-gamma during or after infection. Furthermore, addition of IFN-gamma together with MHV did not prevent infection, but appeared to prevent subsequent viral spread. MHV variants with different degrees of neurovirulence in mice had correspondingly different levels of sensitivities to IFN-gamma treatment in vitro, with the most virulent strain being most resistant to IFN-gamma treatment. Infection of susceptible mice with DE-IFN-gamma-containing virus caused significantly milder disease, accompanied by more pronounced mononuclear cell infiltrates into the CNS and less virus replication, than that caused by virus containing a control DI vector. This study thus demonstrates the feasibility and usefulness of this MHV DI vector for expressing cytokines and may provide a model for studying the role of cytokines in MHV pathogenesis. PMID- 9217057 TI - Bovine immunodeficiency virus tat gene: cloning of two distinct cDNAs and identification, characterization, and immunolocalization of the tat gene products. AB - cDNAs encoding the bovine immunodeficiency virus (BIV) transactivator gene (tat) were cloned from virally infected cells and characterized. BIV expresses two distinct tat mRNAs composed of three exons that are derived by alternative splicing. The BIV tat mRNA splice variants encode Tat proteins of 103 (Tat103) and 108 (Tat108) amino acids. The Tat103 coding region is specified only by exon 2, while that of Tat108 is specified by a truncated exon 2 and the first 30 nt of exon 3. Thus, the first 98 amino acids of each Tat are identical, and have amino terminal, cysteine-rich, conserved core, basic, and carboxyl-terminal domains similar to Tats encoded by primate lentiviruses. BIV-infected bovine cells express a 14-kDa phosphorylated Tat protein identical in size to recombinant Tat expressed in bacteria. BIV Tat was shown to localize exclusively in the nucleoli of virally infected and Tat-expressing cells. Reporter gene assays indicated that Tat103 and Tat108 can strongly transactivate the BIV long terminal repeat (LTR) in virally permissive canine Cf2Th and nonpermissive HeLa and mouse NIH 3T3 cells, but not in permissive lapine EREp cells. However, an intact BIV tat gene is required for viral replication in both Cf2Th and EREp cells. Strong LTR activation by BIV Tat requires a TAR (transactivation responsive) element delimited by viral nt +1 to +31 and the Tat basic domain. BIV Tat strongly cross transactivates the HIV-1 LTR in a TAR-dependent manner in Cf2Th, but not in EREp, HeLa, or NIH 3T3 cells. In contrast, strong, TAR-dependent cross-transactivation of the BIV LTR by HIV-1 Tat could not be demonstrated in any of these cell types. In Cf2Th cells Tat108 effects a moderately stronger transactivation of the BIV LTR than Tat103, indicative of a functional difference in BIV Tat proteins encoded by the mRNA splice variants. The present studies demonstrate that BIV Tat parallels the primate lentiviral Tats in structure and biochemistry but is not interchangeable with the latter. PMID- 9217059 TI - Modulation of stress protein (hsp27 and hsp70) expression in CD4+ lymphocytic cells following acute infection with human immunodeficiency virus type-1. AB - This study was designed to assess the impact of acute human immunodeficiency virus (HIV-1) infection on host intracellular expression of the heat shock family of stress proteins (hsps). Experimental conditions were established wherein CD4+ lymphocytic cell lines undergo a synchronous HIV-1 infection cycle. During the early phase of infection, HIV-1 mRNA expression was restricted to singly and multiply spliced subspecies, with no genomic viral RNA present until 30 hr following infection. In contrast, hsp27 and hsp70 mRNA transcription appeared as early as 3-8 hr following viral infection. No corresponding induction was observed in mock-infected cells. Notably, hsp27 and hsp70 mRNA transcripts were down-regulated by 24 hr, concomitant to the first appearance of full-length genomic HIV-1 mRNA. Hsp27 and hsp70 mRNA transcripts reemerged at end stages of the viral replicative cycle, coincident to virion release and CD4 cell death. Similarly, a transient induction of de novo hsp27 protein expression occurred between 12 and 24 hr. The generated hsp27 stress response was viral dose-related, suppressed by heat-inactivation of virus, and abrogated by neutralizing antibodies to HIV-1. Acute infection did not alter levels of hsp60, hsp70, and hsp90 protein synthesis. However, two-dimensional Western blot analysis did show the appearance of novel hsp70 homologues between 6 and 24 hr following infection. CEM.NKR, Jurkat, H9, and MT-2 cells showed similar patterns of viral-associated modulation of host hsp27 and hsp70 protein and RNA expression. Thus, host hsp27 and hsp70 stress pathways are selectively implicated in the HIV-1 viral life cycle. PMID- 9217058 TI - The murine cytomegalovirus (MCMV) homolog of the HCMV phosphotransferase (UL97(pk)) gene. AB - The murine cytomegalovirus (MCMV) M97 gene is homologous with both eukaryotic protein kinases and the phosphotransferases of herpesviruses. The gene conserves the domain structure of protein kinases and of the human cytomegalovirus UL97 (phosphotransferase) gene. An M97 transcript of 2.5 kb is present predominantly at late times, and much smaller quantities of the transcript are detected at early times postinfection. Comparison of the DNA sequences of the complete M97 genes from 12 ganciclovir-sensitive and aciclovir-sensitive strains of MCMV showed that the sensitive isolates strongly conserve the sequence of the catalytic domains, but have only moderate conservation of the sequence of the amino-terminal (regulatory) region. MCMV provides a useful model for studying the in vivo function of the phosphotransferase genes of the betatherpesviruses and has potential for use in studies of antiviral resistance. PMID- 9217060 TI - Biologic properties of hepatitis B viral genomes with mutations in the precore promoter and precore open reading frame. AB - It is now well recognized that mutations in the hepatitis B virus (HBV) genome occur during the natural course of chronic viral infection. Regions of the viral genome that are frequently affected by such mutations, rearrangements, and/or deletions generally involve the precore promoter, precore, and core as well as the preS gene regions. However, little is known regarding the biologic consequences of these mutations on the functional properties of the variant viral strains with respect to effects on viral replication. In this study, we investigated the functional significance of precore promoter and precore gene mutations that reduce or abolish the synthesis of hepatitis B e antigen (HBeAg). We found that precore promoter mutations diminished the expression of HBeAg but did not affect the synthesis of pregenomic RNA. However, these precore mutations were associated with a modest increase in HBV replication. In contrast, a naturally occurring mutant that carries a termination codon in position 28 of the precore open reading frame demonstrated increased encapsidation of pregenomic mRNA into nucleocapsid particles. Consequently, this variant viral strain demonstrated a substantial increase in the level of viral replication compared to "wild-type" HBV and other precore promoter mutant viral strains. These studies suggest that substitutions in the precore promoter and precore gene not only alter the synthesis of HBeAg but also affect the level of viral replication. PMID- 9217061 TI - Varicella-zoster virus glycoproteins E and I expressed in insect cells form a heterodimer that requires the N-terminal domain of glycoprotein I. AB - Varicella-zoster virus (VZV) glycoproteins E and I (gE and gI), which are major components of the virion envelope, form a noncovalently linked complex. To understand their properties and functions, we expressed and purified soluble forms of gE and gI in the baculovirus system. Extracellular domains of gE and gI were cloned into baculoviruses, using either native or insect-derived signal peptides. Each recombinant virus yielded soluble protein in culture medium although a higher level of secretion was achieved with insect-derived signal peptides in recombinant gE baculoviruses. A soluble gE-gI complex was formed by co-infecting insect cells with recombinant gE and gI baculoviruses and detected by immunoprecipitation followed by Western blotting analyses. By gel filtration and cross-linking studies, we showed that the VZV gE-gI complex expressed in insect cells is a heterodimer. Interestingly, two recombinant gI proteins in which signal peptides were replaced with insect-derived signal peptides did not associate with gE. Amino-terminal sequencing and site-specific mutational studies showed that the replacement of only the signal peptides did not prevent complex formation but alterations in the processed amino-terminus of gI abrogated its ability to complex with gE. These findings indicate that the mature amino terminus of gI is required for gE-gI complex formation by the external domains of VZV gE and gI. PMID- 9217062 TI - The elevation of cellular phosphatidic acid levels caused by polyomavirus transformation can be disassociated from the activation of phospholipase D. AB - Middle T (mT), the oncogene of murine polyomavirus, causes transformation of rat fibroblasts by activating a number of signal transducing pathways usually used by polypeptide growth factors and their receptors. Here, we report data regarding the activation of signal transducing pathways involving phospholipase D (PL-D). The hydrolysis of phospholipids by PL-D produces phosphatidic acid (PA), a compound with multiple biological effects. The PA content of cells expressing wild-type mT, introduced via a number of different methods, is approximately 50% higher than their untransformed counterparts. This increase in cellular PA content is associated with an approximately 65% increase in PL-D activity in cells expressing wild-type mT. We have also examined the effects of a number of site-directed mutants of mT, on both cellular PA levels and on PL-D activity. Mutants that do not produce mT (Py808A) or that produce a truncated, nonmembrane bound mT (Py1387T) have PA levels similar to that of control cells. Cells expressing the 322YF mutant of mT (which abolishes interaction of mT with phospholipase C gamma1) show increases in both PA levels and PL-D activity that are similar to those seen with wild-type mT. Expression of mutants that abolish the interaction of mT with either shc or with phosphatidylinositol 3-kinase (250YS and 315YF, respectively) cause an increase in PL-D activity comparable to that seen with wild-type mT. However, the PA content of cells expressing these mutants is not elevated. These results suggest that mT causes activation of cellular PL-D, but this activation alone is not sufficient to cause an increase in cellular PA content. Therefore, wild-type mT must affect another, as yet unknown, step in PA metabolism. PMID- 9217063 TI - Adaptation of egg-grown and transfectant influenza viruses for growth in mammalian cells: selection of hemagglutinin mutants with elevated pH of membrane fusion. AB - A series of eight transfectant influenza viruses was generated by reverse genetics for studies of the palmitylated cysteine residues in the cytoplasmic tail of the hemagglutinin glycoprotein (HA). Following amplification of these viruses in MDCK cells we found that all had developed an elevated pH of membrane fusion--an unexpected result since previous mutant HA expression studies had shown that substitutions of the cysteine residues had no effect on fusion properties. Sequence analyses revealed that each of the viruses had at least one additional mutation in the ectodomain of HA which was responsible for the increase in fusion pH. Similarly, when we passaged egg-grown wild-type X-31 virus in three different lines of MDCK cells or in MDBK cells, high pH fusion mutants were selected within a few passages in every case. The locations of the substitutions in the HA structure are in or near the "fusion peptide" or at subunit interfaces throughout the length of the trimer--reminiscent of the changes selected in earlier studies on amantadine resistance. The observation that passage of certain viruses in mammalian cells can result in the selection of mutants with elevated fusion pH has potential implications both for reverse genetic experiments and, perhaps more importantly, for the choice of substrates for propagation of vaccine viruses. PMID- 9217064 TI - Unique sequence and lesional tropism of a new variant of neuropathogenic friend murine leukemia virus. AB - FrC6 murine leukemia virus (MuLV) is a replication-competent, neuropathogenic variant derived from Friend MuLV (F-MuLV) complex. The A8 virus (a molecular clone of the FrC6 virus) induced marked spongiform degeneration in the brain similar to the FrC6 virus, but only mild lesions were found in the spinal cord. In contrast, PVC211 virus, which is also a neuropathogenic F-MuLV variant, caused marked spongiform degeneration in the spinal cord. Virus recovery from the spinal cord of A8 virus-infected rat was the same as that of PVC211-infected rat, indicating that there is no direct correlation between the titer of virus and the intensity of lesions. Furthermore, rats infected with the A8 virus at 3 weeks of age did not undergo spongiform degeneration, although recovery of high titer of virus occurred in the central nervous system (CNS). Studies using chimeric viruses between the A8 virus and nonneuropathogenic F-MuLV clone 57 also indicated that the sequences responsible for virus titers in the CNS and neuropathogenicity are different. The chimeric virus studies proved that the env gene and the LTR and/or 5' leader sequence of A8 are critical for the induction of neuropathogenicity. These sequences in A8 and PVC211 were compared, focusing in on the sites that account for neurovirulence and viral lesional tropism. PMID- 9217065 TI - Cloning and sequencing of the cellular-viral junctions from the human adenovirus type 5 transformed 293 cell line. AB - The human-viral junctions of integrated adenovirus type 5 (Ad5) DNA in 293 cells have been cloned and sequenced. The Ad5 sequences extend from nucleotides (nt) 1 to 4344 and are located in the pregnancy-specific beta-1-glycoprotein 4 (PSG 4) gene. This maps the insertion of Ad5 DNA to human chromosome 19 (19q13.2). The Ad5 sequences are represented as a single collinear insertion of viral DNA with no rearrangements. There were 19 bp of PSG4 DNA deleted at the site of insertion and an extra 3 nt (GTC) at the left viral/cellular junction. A short patch of homology between viral DNA and PSG4 DNA (6 nt with 1 mismatch) was present at the right junction. PMID- 9217066 TI - Turnip yellow mosaic virus RNA-dependent RNA polymerase: initiation of minus strand synthesis in vitro. AB - An RNA-dependent RNA polymerase (RdRp) activity was detergent-solubilized from the chloroplast membranes of Chinese cabbage leaves infected with turnip yellow mosaic virus (TYMV). The template-dependent, micrococcal nuclease-treated activity synthesized full-length minus strands from TYMV RNA and 3'-fragments as short as a 28-nucleotide-long RNA comprising the amino acid acceptor stem of the 3'-tRNA-like structure (TLS). Minus strands were shown to arise by de novo initiation with the insertion of GTP opposite the penultimate (C) residue of the 3'-terminal -CCA. The TYMV RdRp activity was template specific in that poly(A) RNA was not copied, and alfalfa mosaic virus (AIMV) RNA, which does not contain a 3'-TLS, was a very poor template. However, other viral RNAs with a 3'-TLS and in vitro transcripts of tRNAs were copied to varying degrees. Fully modified tRNAs were either inactive or poorly active templates, and AIMV 3'-RNA, even when provided with a 3'-terminal -ACCA, was not copied detectably. A potential role of the acceptor stem pseudoknot as a promoter element was assessed with mutations that drastically altered the structure and sequence of the pseudoknot, revealing only a twofold effect in decreasing template activity. The data show that RNAs with both a tRNA-like conformation and a -CCA 3'-terminus are potential templates for TYMV RdRp and suggest that promoter elements are not limited to the acceptor stem pseudoknot. PMID- 9217067 TI - Roles of N-glycans with alpha2,6 as well as alpha2,3 linked sialic acid in infection by polyoma virus. AB - Specific glycosylation inhibitors were used to show that N-glycans, and not O glycans or glycolipids, constitute the major class of receptors on 3T3 cells for polyoma virus. Sialic acid (SA)-specific lectins were used in attempts to confirm the expected SA linkage on N-linked glycans. A lectin specific for alpha2,6 linked SA, a non-receptor type, was slightly more effective in blocking infection than a lectin specific for alpha2,3 linked SA which corresponds to the known receptor specificity. Possible explanations are suggested for this unexpected result. PMID- 9217068 TI - Pharmacological activities of glial cell line-derived neurotrophic factor (GDNF): preclinical development and application to the treatment of Parkinson's disease. PMID- 9217069 TI - Local changes in vascular architecture following partial spinal cord lesion in the rat. AB - Lesions of the CNS induce a complex cascade of tissue reactions. The purpose of this study was to determine the response of the vasculature to partial spinal cord transection. Adult rat spinal cords were lesioned and then examined during acute, subacute, and chronic periods for the presence of endothelial cells and blood vessels at the lesion site. The association of endothelial cells and astrocytes was examined immunohistochemically (RECA-1 and glial fibrillary associated protein, respectively). During the first 48 h following an incision lesion of the dorsal spinal cord, the vasculature was significantly decreased, concurrently with the tissue loss due to primary and secondary degeneration. Subsequently, at 4 days postlesion, vasculature repair processes were evidenced by a significant increase in the number of vessels present at the lesion center. Blood vessels even formed in areas densely packed with macrophages and tissue debris. After 1 week, the number of blood vessels declined in the lesion center and at the place of the forming caverns. These results show significant initial attempts at repair of the vasculature which do not, however, lead to the restoration of a compact tissue and cannot prevent the subsequent formation of caverns. PMID- 9217070 TI - Regulation by interleukin-1beta of formation of a line of delimiting astrocytes following prenatal trauma to the brain of the mouse. AB - The regulation of perinatal glia limitans (GL) reformation by interleukin-1beta (IL-1beta) following prenatal neural trauma in the mouse was studied in lesioned fetal mice by immunocytochemistry and computer-assisted image analysis for presence and distribution of astrocytes and IL-1beta immunoreactivity (ir). Astrocytes stained with anti-glial fibrillary acidic protein (GFAP) were observed as a line of delimiting astrocytes (LDA) near the lesion edge on Postnatal Day 0 (P0, 2 days postlesion). At P6, a new and complete GL composed of GFAP-positive astrocytes was continuous with that of adjacent undamaged tissue. The new GL was located in the same area at P6 as was the LDA at P0, suggesting that the LDA is the precursor structure to a reformed GL. Astrocytes comprising the new GL were positive for anti-IL-1beta. The IL-1 receptor antagonist (IL-1ra), administered acutely into the lesion, produced a significantly decreased optical density of IL 1beta-ir at the LDA at P0 compared to animals that received injections of vehicle, human recombinant IL-1beta, or a combination injection of IL-1ra + IL 1beta. Furthermore, although GFAP-stained cells appeared at the lesion site, an organized LDA was not visible at P0 in IL-1ra-treated animals. Vehicle-, IL-1beta , and combination-injected animals showed a robust LDA at the lesion site at P0. These data suggest that upregulation of IL-1beta in astrocytes and interaction of IL-1beta with the neural IL-1 receptor are important for reconstruction of the GL following prenatal lesion in the murine brain. PMID- 9217072 TI - Age-dependent induction of nitric oxide synthase activity in facial motoneurons after axotomy. AB - The facial nerve was transected in rats at different postnatal ages, from birth to early adulthood. NADPH-diaphorase histochemistry was performed to analyze the induction of nitric oxide synthase, the synthetic enzyme of the free radical nitric oxide, in injured facial motoneurons. In addition, in situ nick-end labeling of DNA fragmentation (TUNEL technique) was performed after axotomy at birth, to verify the occurrence of apoptosis in the damaged facial motoneurons. A striking age-dependency was found in the induction of nitric oxide synthase activity in axotomized facial motoneurons. NADPH-diaphorase positivity was not detectable in these neurons 1 and 2 days after axotomy at birth, when apoptotic changes were evident and marked. In addition, NADPH-diaphorase staining was hardly detectable in the facial nucleus 4 days after axotomies at birth, when extensive motoneuron loss was evident. NADPH-diaphorase positivity was instead induced in the facial motoneurons axotomized from the end of the first postnatal week to adulthood, when the nerve cell loss was less severe than in newborns. However, the time course of the enzyme activity induction varied considerably in relation to the animals' age. These findings are discussed in relation to the role of nitric oxide in motoneuron death or protective response to injury and of oxidative stress in neurodegeneration. PMID- 9217073 TI - Cardiopulmonary interactions following REM sleep deprivation in Sprague-Dawley rats. AB - We characterized the effects of 48 h of rapid-eye-movement (REM) sleep deprivation on cardiovascular and respiratory variables and on sleep-related cardiopulmonary interactions in adult male Sprague-Dawley rats. Rats were instrumented for monitoring EEG, EMG, and aortic blood pressure. Respiratory rate and minute ventilation were measured by unrestrained single-chamber plethysmography. By using radiotelemetry to monitor blood pressure we clearly demonstrated progressive decreases in mean blood pressure with transitions from wakefulness to non-rapid-eye-movement and REM sleep which were unaffected by REM sleep deprivation. Mirror-image state-dependent increases in heart period suggest that baroreflexes were augmented during sleep with respect to wakefulness. REM sleep deprivation was also associated with lower blood pressure and longer heart period over all sleep/wake states, although this achieved statistical significance only during REM sleep and only during the first hour of recovery sleep. These cardiovascular changes coupled with the observed decreases in respiratory rate and minute ventilation suggest a further augmentation of baroreflexes following REM sleep deprivation. PMID- 9217071 TI - Survival, integration, and differentiation of neural stem cell lines after transplantation to the adult rat striatum. AB - The in vivo properties of four different neural stem cell lines, generated from embryonic striatum or hippocampus by immortalization with the temperature sensitive (s) A58/U19 allele of the SV40 Large T-antigen, have been studied with respect to their ability to survive, differentiate, and integrate after transplantation to the adult rat striatum. The cells were labeled with [3H]thymidine prior to grafting, and combined autoradiography and immunohistochemistry was used to characterize their phenotypic differentiation within the adult brain environment. The results show that all four types of cells survived well, up to at least 1.5-6 months postgrafting, without any signs of tissue perturbation or tumor formation. The cells underwent, on average, 2-3 cell divisions during the first 5 days after implantation and exhibited extensive migration over a distance of 1-1.5 mm from the injection site to become morphologically integrated with the surrounding host striatum. The cell number and tissue distribution attained by 2 weeks remained stable for up to 6 months postgrafting with the exception of one cell line, which showed a 40% loss of cells between 2 and 6 weeks. Twice the number of [3H]thymidine-labeled cells were recovered when the cells were grafted into a 1-week-old excitotoxic striatal lesion, probably due to an increased proliferation of the cells in response to the neuron-depleting depleting lesion. The immortalized cells behaved as multipotent neural progenitors. The vast majority of the cells developed a glial like morphology, 6-14% being clearly GFAP-positive; however, a small but consistent proportion of them (1-3%) expressed MAP-2 and exhibited neuron-like morphology. In mature transplants about 75-80% of the grafted cells were located in the striatal grey matter, and 10-15% in white matter, some of which are proposed to have differentiated into oligodendrocytes. Remaining 5-10% occurred around small blood vessels (resembling pericytes) and in the subventricular zone underneath the ependyma of the lateral ventricle. It is concluded that the ts cell lines are highly suitable for intracerebral transplantation and that they allow the creation of a regionally confined cellular chimeras where the graft derived glial cells become stably integrated with the resident glial cell matrix. PMID- 9217074 TI - The responses of mammalian spinal axons to an applied DC voltage gradient. AB - We have imposed a steady, rostrally negative, weak (ca 0.4 mV/mm) voltage gradient across transections of ascending white matter tracts in the adult guinea pig using an implanted stimulator and electrodes for about 1 month. We have evaluated the projections of these axons relative to the transection approximately 2 months postinjury by anterograde transport of injected tetramethylrhodamine-conjugated dextran and the use of an indwelling marker device which locates the plane of the original transection. Tract tracing was accomplished with conventional epifluorescence microscopy and confocal laser microscopy. Sham-treated control spinal cords contained well-filled lateral and dorsal column ascending tracts terminating caudal to the lesion which formed at the level of the hemisection. Electric field-treated spinal cords contained similarly labeled columns of axons that penetrated the lesion within the caudal segment of the spinal cord, branched within it, and in some cases such branches projected across the plane of transection. Ascending axons also passed around the lesion through undamaged parenchyma, branched repeatedly at the plane of the hemisection, and passed into the rostral segment of the spinal cord. Spear-shaped endings typical of growth cones were found at the terminals of these processes which often branched again within the rostral segment. Centrally projecting fibers, their processes, and the overall level of branching in these projections was not observed in our previous studies using high molecular weight horseradish peroxidase tracers. PMID- 9217075 TI - Effect of perinatal asphyxia on systemic and intracerebral pH and glycolysis metabolism in the rat. AB - The effects of perinatal asphyxia on systemic and brain pH and glycolysis metabolism were studied in the rat. Perinatal asphyxia was induced by immersing pup-containing uterus horns, obtained by cesarean section from rats within the last day of gestation, in a water bath at 37 degrees C for various periods of time (0-23 min). Subcutaneous levels of pyruvate (Pyr), lactate (Lact), glutamate (Glu), and aspartate (Asp) were monitored with microdialysis 40-80 min after delivery. In parallel experiments, the pups were sacrificed 40 min after delivery and the heart and brain were removed for measuring pH. Brain (striatum) Pyr, Lact, Glu, and Asp levels were also analyzed. A decrease in the rate of survival was first observed following asphyctic periods longer than 16 min, and no survival could be observed after 22 min of asphyxia. In control (cesarean delivered) pups, heart and brain pH were 7.36 +/- 0.01 (N = 8) and 7.30 +/- 0.01 (N = 8), respectively. Significant decreases in pH were first observed following 5-6 and 10-11 min of asphyxia, in heart and brain, respectively. In both regions pH decreased along with the length of asphyxia, but a decrease below 7 was only observed in the brain, following asphyctic periods longer than 16 min. A significant increase in subcutaneous Lact levels was first observed following 2-3 min of asphyxia, with a maximum after 20-21 min of asphyxia. In the brain, the increase in Lact levels was delayed compared to that observed in subcutaneous tissue. Pyr and Asp levels increased in subcutaneous tissue following perinatal asphyxia and decreased in brain tissue following > 15 min of asphyxia. Glu levels were increased subcutaneously by moderate (5-16 min) asphyctic periods, but, in the brain, were only transiently increased by 10-11 min of asphyxia. Thus, changes in systemic pH, glycolysis, and excitatory amino acid metabolism are observed following shorter asphyctic periods than are changes in the brain. In particular, increases in subcutaneous Lact levels precede: (i) a decrease in brain pH, (ii) an increase in brain Lact levels, (iii) a decrease in the rate of survival, and, probably, (iv) brain damage. It is suggested that monitoring Lact levels by subcutaneous microdialysis is a useful method for predicting the outcome produced by hypoxic-ischemic insults. PMID- 9217076 TI - Integration of transplanted cultured Schwann cells into the long myelinated fiber tracts of the adult spinal cord. AB - A suspension of about 10,000 purified Schwann cells cultured from the neonatal rat sciatic nerve was transplanted into a discrete site in the upper cervical level of the corticospinal tract of one side in adult rats. From 4 days after transplantation immunostaining for p75 (low-affinity neurotrophin receptor) showed that the transplants consisted of a central mass of Schwann cells and cuffs of elongated Schwann cells along the perivascular space of curving blood vessels (most of which had been formed in response to the transplantation). Schwann cells leaving the central mass and perivascular cuffs migrated in strictly linear orientation along the rostrocaudal axis of the host corticospinal tract. According to the territory through which they migrated, the transplanted Schwann cells adopted two quite different forms: (1) The row Schwann cells, which migrated singly or in groups within the rows of host oligodendrocytic and astrocytic cell bodies, were non-process-bearing, rather cuboidal, brick-like cells (about 8 x 12 microm in size). (2) In contrast, the interfascicular Schwann cells, which migrated singly or intertwined in rope-like small groups interspersed among the axons of the host corticospinal tract, were larger, symmetrically bipolar cells, with processes about 100-120 microm long and 2 microm wide and bulging, ovoid nuclei, located in centrally placed cell bodies about 10 microm across. After about 6 weeks, the p75 immunoreactivity of the interfascicular Schwann cells had become down-regulated. However, from as early as 10 days after transplantation, immunostaining for the peripheral myelin protein, P0, semithin sections, and electron microscopy showed that these Schwann cells were not lost, but that they had myelinated the segments of the host corticospinal axons in the region of the transplant. In contrast, the row Schwann cells did not express P0 or form myelin. They retained their p75 immunoreactivity at long survivals (presumably because they were secluded from contacting the tract axons). The row Schwann cells also migrated farther than the interfascicular Schwann cells (possibly a function of their maintained p75 expression), becoming dispersed singly for at least 8 mm from the original transplant site. Our previous study of corticospinal tract lesions had shown the formation of a "closed" scar formed by hypertrophic astrocytic processes, which walled off a central astrocyte-free region and totally disrupted the normal longitudinal alignment of the tract astrocytic processes. In contrast, while the present Schwann cell transplants induced a comparable astrocytic hypertrophy over the same time course, the astrocytic processes remained able to penetrate the transplant site, which was not walled off, so that the longitudinal arrangement of the host corticospinal tract astrocytic skeleton was preserved intact across the region of the transplant. These observations show that Schwann cells can be intimately integrated into the cytoarchitecture of the myelinated adult host corticospinal tract. This integration is not a random dispersal in damaged areas: it involves direct interaction with the cell elements present in the host tract, it respects the complex and regular organization of the host tract glial cells, and it results in the formation of a precisely arranged mosaic of central and peripheral tissue. PMID- 9217077 TI - Expression of NMDA receptor-1 (NR1) and huntingtin in striatal neurons which colocalize somatostatin, neuropeptide Y, and NADPH diaphorase: a double-label histochemical and immunohistochemical study. AB - The subset of striatal neurons which colocalize SS/NPY/NADPH-d are selectively resistant to neurodegeneration in Huntington's Disease (HD) and to excitotoxic cell death induced experimentally with NMDA receptor (NMDAR) agonists. Here we have analyzed the expression of immunoreactive NMDAR-1 (NR1) subunit (as an index of NMDAR protein) and of huntingtin (the normal product of the HD gene) in primary cultures of rat striatum to see if differential expression of the two antigens in the subset of SS/NPY/NADPH-d and other striatal neurons can explain their selective resistance or vulnerability. Double-label histochemical and immunocytochemical studies were carried out using conventional and confocal laser scanning microscopy to characterize the cellular and subcellular expression of NR1 and SS, or NPY or bNOS, together with NADPH-d histochemistry. The percentages of cultured striatal neurons that were positive for NADPH-d, SS, NPY, bNOS, and NRI were, respectively, 3.8, 8.4, 10.2, 5.1, and 80%. The majority of striatal NADPH-d neurons coexpressed SS and NPY; 17% of SS-producing neurons were strongly positive for NR1; the remaining cells (approximately 80%) exhibited only weak NR1 expression. Comparable data were obtained for NPY-positive neurons, 15% of which colocalized NR1 strongly and 70-80% weakly. By double-label immunofluorescence, huntingtin was nonselectively expressed in virtually all striatal neurons including SS/NPY/NADPH-d neurons. These results show that the majority of striatal SS/NPY/NADPH-d neurons express NR1. The relative abundance of NR1 in SS/NPY/NADPH-d neurons, however, varies between a small subset of neurons that are receptor rich and the remainder that express low levels only and may determine susceptibility to NMDAR-mediated neurotoxicity. Huntingtin is nonselectively expressed in virtually all striatal neurons and does not appear to be a determinant of the selective resistance of normal striatal SS/NPY/NADPH-d neurons to NMDA toxicity. PMID- 9217078 TI - The glycine antagonist GV150526 protects somatosensory evoked potentials and reduces the infarct area in the MCAo model of focal ischemia in the rat. AB - The neuroprotective activity of the novel, selective glycine antagonist GV150526 was assessed in the middle artery occlusion (MCAo) model of focal ischemia. Postischemia administration of GV150526 (3 mg/kg i.v.) up to 6 h post-MCAo resulted in a significant reduction of the infarct volume measured histologically 24 h later. The neuronal protection by GV150526 was accompanied by functionally significant protection determined by somatosensory evoked potential (SEP) responses recorded from the primary somatosensory cortex of rats under urethane anesthesia. Experimental occlusion of the MCA 7 days prior to electrophysiological testing induced a clear reduction in the SEP amplitude. GV150526 (3mg/kg, i.v.) was able to protect SEP responses recorded from the hindpaw cortical field in two groups of animals treated either 1 (n = 9) or 6 h (n = 10) post-MCAo. SEP responses recorded from the forepaw cortical field, an area closer to the core of the ischemic damage, were significantly protected only in the group treated 1 h post-MCAo. Histological evaluation of the rat brain regions showed a correlated decrease in the ischemic area of GV150526-treated groups. The volumes of the ischemic brains of both GV150526 groups were statistically different from the MCAo group (P < 0.05). These findings demonstrate that GV150526 is able to prevent the ischemic damage assessed histologically and affect the functional correlates of the ischemia evaluated by the electrophysiological SEP measurements. PMID- 9217079 TI - Catecholaminergic development of fetal rat ventral mesencephalon: characterization by high-performance liquid chromatography with electrochemical detection and immunohistochemistry. AB - We determined dopamine (DA), noradrenaline (NA), and adrenaline (A), as well as immunohistochemically stained tyrosine hydroxylase (TH) and DA in dissected rat ventral mesencephalon (VM) tissue from Embryonic Day (ED) 14 to Postnatal Day (P) 17. Whole VM tissue DA, NA, and A contents increased with advancing age. VM DA/protein increased from ED15 to ED16, whereas NA/protein increased from ED15 to ED16 and from ED20 to P4. VM DA/NA ratio increased from ED14 to ED15 and decreased from ED18 to P4. VM cell suspensions exhibited higher DA/NA ratios than whole VM tissue. Washed cell suspensions had higher DA/NA than unwashed counterparts. We conclude that data from both VM immunohistochemistry and catecholamine assays relate to VM development. VM DA is contained mainly in cells, whereas VM NA is located in fibers that channel at the dorsal side of the VM. Determination of tissue catecholamine contents may be helpful for the biochemical characterization of tentatively identified VM grafts. PMID- 9217080 TI - Anticonvulsant action of both NMDA and non-NMDA receptor antagonists against seizures induced by homocysteine in immature rats. AB - Seizures were induced in immature 18-day-old rats by i.p. administration of homocysteine (11 mmol/kg) and the effects of selected antagonists of NMDA receptors [MK-801 (0.5 mg/kg), AP7 (0.33 mmol/kg), CGP 40116 (10 mg/kg)] and non NMDA receptors [GDEE (4 mmol/kg), NBQX (two doses, 30 mg/kg each)] were studied. The effect of MgSO4 (two doses, 2 mmol/kg each) was also tested. The anticonvulsant effect was evaluated not only from the behavioral manifestations of seizures, but also in terms of some indicators of brain energy metabolism. Rat pups were sacrificed during generalized clonic-tonic seizures, corresponding to 16-45 min after homocysteine administration. Comparable time intervals were used for sacrificing the pups which had received the protective drugs. In contrast to neonatal rats, in which only NMDA antagonists could prevent homocysteine-induced seizures, both NMDA and non-NMDA receptor antagonists exerted an anticonvulsant effect in 18-day-old rats. In addition, the pronounced anticonvulsant effect could be achieved by the combined treatment with low subthreshold doses of NMDA (MK-801) and non-NMDA (NBQX) receptor antagonists. The protection was evident not only in suppressing behavioral symptoms of seizures, but also in preventing most of the metabolic changes accompanying seizures, mainly glycogen degradation. More than a sevenfold accumulation of lactate occurring during seizures was markedly reduced by all the tested drugs, but was not completely eliminated. All antagonists, when given alone in the same doses as those used for seizure protection, remained without any effect on lactate levels. Comparison of the present data with previous findings concerning neonatal rats suggests that there may be a developmental change in anticonvulsant efficacy of non-NMDA receptor antagonists against homocysteine-induced seizures in rats. PMID- 9217081 TI - Immediate early gene expression and delayed cell death in limbic areas of the rat brain after kainic acid treatment and recovery in the cold. AB - Systemic injection of kainic acid (KA) results in characteristic behaviors and programmed cell death in some regions of the rat brain. We used KA followed by recovery at 4 degrees C to restrict damage to limbic structures and compared patterns of immediate early gene (IEG) expression and associated DNA binding activity in these damaged areas with that in spared brain regions. Male Wistar rats were injected with KA (12 mg/kg, i.p.) and kept at 4 degrees C for 5 h. This treatment reduced the severity of behaviors and restricted damage (observed by Nissl staining) to the CA1 and CA3 regions of the hippocampus and an area including the entorhinal cortex. DNA laddering, characteristic of apoptosis, was first evident in the hippocampus and the entorhinal cortex 18 and 22 h after KA, respectively. The pattern of IEG mRNA induction fell into three classes: IEGs that were induced in both damaged and spared areas (c-fos, fos B, jun B, and egr 1), IEGs that were induced specifically in the damaged areas (fra-2 and c-jun), and an IEG that was significantly induced by saline injection and/or the cold treatment (jun D). The pattern of immunoreactivity closely followed that of mRNA expression. Binding to the AP-1 and EGR DNA consensus sequences increased in all three regions studied. This study describes a unique modification of the animal model of KA-induced neurotoxicity which may prove a useful tool for dissecting the molecular cascade that ultimately results in programmed cell death. PMID- 9217082 TI - Neurotoxicity of soluble macrophage products in vitro--influence of dexamethasone. AB - When macrophage conditioned medium is added to neurons in vitro, there is a loss of cell membrane integrity, a loss of cell processes, and a large increase in apoptotic neurons. We tested the influence of a potent anti-inflammatory steroid on the interaction between macrophages and neurons. Dexamethasone was applied to macrophages in culture for 24 h while the culture was stimulated with lipopolysaccharide and hypoxia. Conditioned medium was collected after dexamethasone was removed. The dexamethasone pretreated medium was not toxic to hippocampal neurons in contrast to medium from stimulated macrophages not treated with steroid. The dexamethasone effect was concentration dependent. Pretreatment of macrophages with indomethacin and transforming growth factor beta had similar but less impressive effects when compared to dexamethasone. The effect of dexamethasone may have been mediated by inhibiting the synthesis or release of neurotoxic macrophage protein(s), as a combination of medium from steroid pretreated macrophages with medium from nontreated macrophages was not neuroprotective. The toxin(s) did not appear to be tumor necrosis factor alpha or arginase. A role for most neutral proteases was also excluded. We also assessed the consequence of stressing neurons with a mild hypoxic exposure immediately prior to conditioned medium application. Medium from dexamethasone-treated macrophages did not exaggerate hypoxic neuronal injury, unlike medium from non dexamethasone-treated macrophages. It did not, however, block the exaggerating effect when coapplied in equal volume with medium from nontreated macrophages. Dexamethasone at 100 nM had no impact when applied directly to neurons while they were being exposed to conditioned medium. This in vitro protection by dexamethasone may be relevant to the demonstrated benefit of glucocorticoids in selected brain and spinal cord conditions. Suspicion of a potential link between this in vitro finding and in vivo CNS injury justifies an assessment of more specific agents acting on macrophage protein synthesis or secretion. PMID- 9217083 TI - Embryonic tissue induces growth of adult axons from myelinated fiber tracts. AB - Suspensions of late embryonic hippocampal tissue were microinjected so as to be completely enclosed within the myelinated fiber bundles of the adult rat fimbria. Previous studies have shown that the axons from such transplanted neurons readily cross the graft/host interface and extend rapidly through the host fiber tract. The present study shows that the adult axons from the host fiber tract can also cross this interface in the opposite direction and enter the transplants. Biotin dextran tracing shows that the adult host fimbrial axons traverse the embryonic grafts and also form terminal arborizations within the transplants. Electron microscopy of orthograde electron-dense degeneration confirms that these host axons form synaptic terminals accounting for at least 6.6% of the synapses in the neuropil of the transplant. Thus, contact with embryonic nervous tissue can induce elongative growth by the adult fibers in a myelinated central tract. PMID- 9217084 TI - Anticipatory control of manipulative forces in Parkinson's disease. AB - In a previous study we found that subjects with Parkinson's disease (PD) had an impaired capability to initiate and sequence successive movement phases during lifts of small objects using the precision grip, and that they had regular oscillations in the force rates. The present study examined whether these subjects could use anticipatory control, in which the force output is scaled prior to liftoff, based on the object's physical properties. Subjects lifted an instrumented test object between the tips of the thumb and index finger while the employed grip force, load force (vertical lifting force), and corresponding time derivatives were recorded. In the first experiment, the object's weight was varied to assess its influence on the isometric force output. Subjects with PD scaled the isometric force increase according to the object's weight. In another experiment, the weight changed in proportion to the volume to determine whether subjects could make associative transformations between visual size information and the weight of the object. Subjects with PD still scaled the forces toward the expected weight, proportional to the volume of the object. Finally, programmed adjustments in force to sudden self-induced load changes were examined while subjects dropped a disk with one hand into a plate attached to the bottom of the grip instrument, held with the other hand. Subjects with PD had preparatory increases in the grip force prior to the disk contact, which matched the change in load, though may have been more dependent on visual feedback. We conclude that subjects with PD are capable of using anticipatory control to parameterize the isometric force output during a familiar lifting task. PMID- 9217085 TI - Coordination of manipulative forces in Parkinson's disease. AB - The coordination of manipulative forces was examined in 10 subjects with Parkinson's disease (PD) both OFF and ON medication while they grasped and lifted a small object using the precision grip. The development of grip (squeeze) force and load (vertical lifting) force was recorded and compared to a group of age matched control subjects. Subjects with PD often exhibited a prolonged delay between the first digit contact with the object and initiation of the lifting drive. These subjects also exhibited stepwise increases in force, with regular oscillations in the force rates. However, once the vertical drive began, the main increase in grip and load force generally was in parallel and most other temporal aspects of the force coordination were similar to those of the control subjects. The extent to which the movement initiation was delayed was related to the stage of the disease, and most subjects improved ON medication. When the object was held in the air, subjects with PD used a grip force level which was similar to that of the control subjects, and all subjects adjusted their grip force according to the surface texture. Furthermore, they exhibited proper reflexive corrections to sudden changes in load (object perturbations), suggesting intact sensorimotor integration. We conclude that the most obvious impairments in the coordination of this task were delayed initiation of the grip-lift sequence and tremor-like oscillations superimposed on otherwise normal force. PMID- 9217086 TI - Clip compression injury in the spinal cord: a correlative study of neurological and morphological alterations. AB - Rats subjected to experimental spinal cord compression of different degrees induced by aneurysm clips were neurologically tested 3 and 5 weeks postinjury. The development of spinal cord tissue destruction over time was similar to what has been described for other experimental spinal cord injuries with characteristics such as early edema, axonal swelling, and later necrosis. Three weeks after injury a reactive gliosis was found at the injury epicenter and regenerating axons could be identified in the otherwise necrotic cavity. The extent of degeneration was highly correlated with the closing force of the aneurysm clip. The results of a number of neurological tests were correlated to the degree of clip-induced compression, to lesion volume, and to the remaining area of white matter at the epicenter. The neurological tests with the highest correlation to morphological descriptors were beam walk (r(s) = 0.89-0.95) and motor performance score (r(s) = 0.88-0.92). We conclude that the motor performance score, previously validated for photochemically induced ischemic spinal cord injuries, is equally suitable for clip compression injuries as a fast and reliable neurological test paradigm. PMID- 9217087 TI - Astrocytes promote or impair the survival and function of embryonic ventral mesencephalon co-grafts: effects of astrocyte age and expression of recombinant brain-derived neurotrophic factor. AB - Intrastriatal grafting of dopamine-rich embryonic ventral mesencephalon (VM) is a potential therapeutic treatment for Parkinson's disease. However, it has been suggested that the efficacy of this procedure might be improved by enhancing the survival and/or degree of neurite outgrowth by the grafted VM, since these parameters are currently suboptimal. In the present study, we tested the ability of astrocytes retrovirally transduced to produce recombinant brain-derived neurotrophic factor (BDNF) to enhance the survival and/or function of embryonic VM in the unilateral 6-hydroxydopamine (6-OHDA) lesioned rat, a well characterized rodent model of Parkinson's disease. In culture, primary astrocytes derived from Postnatal Day 0 (P0) rat striatum and transduced with the BDNF vector increased the survival of Embryonic Day 15 (E15) dopaminergic VM neurons by approximately threefold and reduced the loss of dopaminergic neurons following 6-OHDA treatment by approximately 20%. The cultured astrocytes were then mixed 1:1 with freshly dissociated E15 VM and co-grafted into the dopamine-denervated striatum. Unexpectedly, the control nontransduced astrocytes reduced the survival of dopaminergic neurons by 60% and restricted the pattern of neurite outgrowth by the co-grafted VM, compared to grafts of VM alone at 7 weeks postgrafting. These effects were paralleled by an attenuated rate and degree of behavioral recovery. The detrimental effects of the control astrocytes were partially reversed when the astrocytes were transduced to express BDNF, although dopaminergic neuron survival was still reduced by 30% compared to that within VM-only grafts. To begin to assess whether the detrimental effects of the astrocytes were related to the maturational state of the cultured astrocytes, astrocytes were obtained from E18 striatum and maintained in short-term culture (9 days vs several weeks for P0 cultures) prior to co-grafting with VM. Interestingly, the younger astrocytes did not reduce graft survival and allowed for better graft integration. These results suggest that primary astrocytes maintained in long-term culture are detrimental to embryonic neural grafts, an effect that is not completely overcome by expression of recombinant BDNF, and that astrocyte age may be an important consideration in the use of these cells as CNS gene delivery vehicles. PMID- 9217088 TI - Collateral sprouting of central noradrenergic neurons during aging: histochemical and neurochemical studies in intraocular triple transplants. AB - The sprouting capacity of aged noradrenergic neurons of the brain-stem nucleus locus coeruleus (LC) was examined using intraocular transplants of fetal tissues. Fetal hippocampal tissue (E18) and LC tissue (E15) were transplanted together as a double transplant into the anterior chamber of the eye of young adult Fischer 344 rats. The double transplants were allowed to mature for 14-18 months, after which an additional fetal hippocampal transplant was placed next to the LC graft. The triple transplants were monitored for overall growth and vascularization for an additional 2-6 months. Immunohistochemical examinations showed that both young (2-6 months old) and aged (16-24 months old) hippocampal cografts contained a plexus of thin varicose tyrosine hydroxylase (TH)-immunoreactive fibers extending throughout the grafted hippocampal tissues. However, the aged hippocampal grafts contained a denser uniform plexus of TH-positive fibers compared to the young transplants. Immunohistochemistry with synapsin antibodies demonstrated that both the young and the aged hippocampal transplants contained much higher densities of synaptic elements than the LC grafts. In vivo electrochemical measurements of potassium-evoked overflow of norepinephrine (NE) in the grafts showed that similar amounts of NE overflow were detected in both the young and the aged hippocampal grafts. HPLC-EC measurements of NE levels in the grafts revealed that there were similar amounts of NE in the young and the aged grafts, and the grafts did not contain serotonin or dopamine. In summary, the findings of the present study show that aged LC neurons are capable of undergoing collateral sprouting producing a functional NE neuronal system when introduced to an appropriate young target. PMID- 9217089 TI - Long-term transgene expression in fetal rat suprachiasmatic nucleus neurografts following ex vivo adenoviral vector-mediated gene transfer. AB - Ex vivo gene transfer to fetal suprachiasmatic nucleus (SCN)-containing solid piece neurografts was explored using a first-generation prototype adenoviral vector containing the reporter gene LacZ (Ad-LacZ). Transgene expression was examined at different intervals following grafting in the IIIrd ventricle of rat brain and was compared to that of explant cultures. Large numbers of beta galactosidase-positive cells were observed 8 days postgrafting. The number of stained cells had decreased considerably at 21 days but transduced cells were still present at 70 days. In vitro culturing of infected SCN tissue revealed high expression up to 21 days, indicating that the in vivo and in vitro fates of Ad LacZ-infected cells were different. The main reason for this difference appeared to be cell loss by necrosis in the initial phase after transplantation, a phenomenon not related to the infection with Ad-LacZ since it similarly occurred in control grafts. In vivo inflammatory responses, observed after immunostaining for macrophages and T-lymphocytes, were also comparable in control and Ad-LacZ treated transplants, except that cytotoxic T-cells were observed in the Ad-LacZ treated transplants and not in controls. The recruitment of these cells was, however, minor and primarily observed at 8 days postgrafting, indicating that a major immunological rejection of the transduced graft did not occur. In both control and Ad-LacZ-infected transplants similar survival and intraimplant neuritic growth of SCN cells were visible. Ex vivo gene transfer of solid piece fetal SCN grafts with adenoviral vectors therefore appeared to be a nontoxic long term gene-introducing procedure. This would in principle enable the local production of neurotrophic factors within the transplant and has the potential to improve functional SCN neurografting. PMID- 9217090 TI - Delivery of recombinant tetanus-superoxide dismutase proteins to central nervous system neurons by retrograde axonal transport. AB - The nontoxic C fragment of tetanus toxin (TC) can transport other proteins from the circulation to central nervous system (CNS) motor neurons. Increased levels of CuZn superoxide dismutase (SOD) are protective in experimental models of stroke and Parkinson's disease, whereas mutations in SOD can cause motor neuron disease. We have linked TC to SOD and purified the active recombinant proteins in both the TC-SOD and SOD-TC orientations. Light microscopic immunohistochemistry and quantitative enzyme-linked immunosorbant assays (ELISA) of mouse brainstem, after intramuscular injection, demonstrate that the fusion proteins undergo retrograde axonal transport and transsynaptic transfer as efficiently as TC alone. PMID- 9217092 TI - Delayed antagonism of AMPA/kainate receptors reduces long-term functional deficits resulting from spinal cord trauma. AB - Excitatory amino acid (EAA) receptors play a significant role in delayed neuronal death after ischemic and traumatic injury to the CNS. Focal microinjection experiments have demonstrated that 2,3-dihydro-6-nitro-7-sulfamoyl benzo(f)quinoxaline (NBQX), a highly selective and potent antagonist of non-N methyl-D-aspartate ionotropic EAA receptors, i.e., those preferring alpha-amino-3 hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) or kainate, can reduce histopathology and functional deficits when administered at 15 min after traumatic spinal cord injury (SCI). Similarly, intravenous infusion of NBQX, beginning at 15 min postinjury (p.i.), results in a significant amelioration of the functional deficits produced by experimental SCI. However, if antagonists of AMPA/kainate receptors were to be used therapeutically for patients with SCI, administration would likely be delayed for several hours after injury. We therefore examined the effects of NBQX administered at 4 h after SCI on functional deficits and histopathology in a standardized rat model of contusive SCI. An incomplete SCI was produced in Sprague-Dawley rats at T8 with a weight drop device (10 g x 2.5 cm). NBQX (15 nmol), or vehicle alone, was microinjected into the injury site 4 h later. Recovery of hind limb reflexes, postural control, and locomotor function was determined by a battery of behavioral tests performed for 8 weeks. Spinal cord tissue was then fixed by perfusion and used for morphometric and immunocytochemical analyses. Previous studies with acute NBQX treatment showed significant functional improvement by 1 week; the effects of delayed NBQX treatment on functional deficits were not discernible until 3-4 weeks after SCI. Thereafter, significant reductions in hindlimb deficits were demonstrated in two independent studies. The nature and magnitude of the reductions in chronic deficits were similar to those observed previously when NBQX was administered acutely at 15 min after SCI. Morphometric analyses showed that delayed treatment with NBQX resulted in sparing of gray matter adjacent to the injury site but no significant effect on the area of white matter at the epicenter. However, serotonin immunoreactivity below the lesion, used as a marker for preservation of one supraspinal pathway, was significantly higher in the NBQX treated group. These results support a therapeutic potential for NBQX, and presumably other AMPA antagonists, in SCI by demonstrating effectiveness in a clinically relevant time frame. They indicate the importance of assessing chronic functional deficits in evaluating the therapeutic potential of a treatment paradigm. Further, they suggest the intriguing hypothesis that mechanisms underlying early functional recovery after SCI are, at least in part, distinct those from those involved in reducing chronic functional deficits. PMID- 9217091 TI - Regional and cellular localization of presenilin-2 RNA in rat and human brain. AB - In situ hybridization probes selective for presenilin-2 (PS-2) were used to determine the regional and cellular expression pattern of PS-2 mRNA in rat and human brain. In rat brain, the greatest expression of PS-2 mRNA is in the granule cell layers of the dentate gyrus and cerebellum. Molecular layers within these structures are virtually devoid of signal. Cortical expression of PS-2 message is restricted to neuronal layers, while the hybridization signal is weak or absent in molecular layers and white matter. Kidney, liver, and spleen display moderate levels of PS-2 message. A PS-2 sense strand probe produced no specific signals in any tissue. In human brain, the greatest hybridization signal for PS-2 is present in the granule cells of the cerebellum. Within hippocampus, the granule cell layer of dentate is strongly labeled, with CA3 pyramidal neurons also clearly visible. A laminar expression pattern is seen in the neuronal layers of human frontal and temporal cortex, with the deeper laminae having the strongest signals. These data are consistent with a primarily neuronal localization of PS-2 mRNA within the brains of both rat and human. Within the limitations of the analysis, it appears that virtually every neuron is labeled, and differences in the intensity of labeling are associated with both neuron size/density and brain region. The distribution of PS-2 RNA is not restricted to those regions having the greatest pathology in Alzheimer's disease. However, one unusual pathological feature of PS-2 mutations causing AD is the presence of cerebellar amyloid plaques in some cases. It is intriguing, in this context, that PS-2 RNA is enriched in the cerebellum, especially in human specimens. PMID- 9217093 TI - Factors determining the size frequency distribution of beta-amyloid (A beta) deposits in Alzheimer's disease. AB - The size frequency distributions of discrete beta-amyloid (A beta) deposits were studied in single sections of the temporal lobe from patients with Alzheimer's disease. The size distributions were unimodal and positively skewed. In 18/25 (72%) tissues examined, a log normal distribution was a good fit to the data. This suggests that the abundances of deposit sizes are distributed randomly on a log scale about a mean value. Three hypotheses were proposed to account for the data: (1) sectioning in a single plane, (2) growth and disappearance of A beta deposits, and (3) the origin of A beta deposits from clusters of neuronal cell bodies. Size distributions obtained by serial reconstruction through the tissue were similar to those observed in single sections, which would not support the first hypothesis. The log normal distribution of A beta deposit size suggests a model in which the rate of growth of a deposit is proportional to its volume. However, mean deposit size and the ratio of large to small deposits were not positively correlated with patient age or disease duration. The frequency distribution of A beta deposits which were closely associated with 0, 1, 2, 3, or more neuronal cell bodies deviated significantly from a log normal distribution, which would not support the neuronal origin hypothesis. On the basis of the present data, growth and resolution of A beta deposits would appear to be the most likely explanation for the log normal size distributions. PMID- 9217094 TI - Levels of L-methionine S-adenosyltransferase activity in erythrocytes and concentrations of S-adenosylmethionine and S-adenosylhomocysteine in whole blood of patients with Parkinson's disease. AB - In the present study, levels of S-adenosylmethionine (SAM) and S adenosylhomocysteine (SAH) in whole blood as well as L-methionine S adenosyltransferase (MAT) activity in erythrocytes were assayed in a series of 20 patients with Parkinson's disease and 12 healthy control subjects. A significant difference was found with regard to SAM levels between patients and controls, with the detected levels being 383.1 +/- 41.5 nM for the parkinsonian patients and 680.6 +/- 30.9 nM for the controls. With regard to SAH, we found no difference between the groups. The catalytic activity of MAT was increased by 30% in patients compared to controls, with the Vmax for methionine being 17.9 +/- 3.7 and 13.9 +/- 2.2 pmol/mg/h, respectively. PMID- 9217095 TI - Cellular delivery of NGF does not alter the expression of beta-amyloid immunoreactivity in young or aged nonhuman primates. AB - The present study determined whether grafts of nerve growth factor-producing fibroblasts alter the expression of beta-amyloid in young or aged nonhuman primates. Aged monkeys serve as an animal model which normally exhibits beta amyloid-laden plaques. Three young adult (7-12 years of age) and three aged (24 29 years of age) rhesus monkeys received intraventricular implants of polymer encapsulated cells that were genetically modified to secrete human recombinant nerve growth factor (NGF). Three young adult and three aged rhesus monkeys received identical treatment except that the grafted cells were not genetically modified and thus differed only by a single gene construct. Five additional aged rhesus monkeys were ungrafted and also served as controls. Three to four weeks posttransplantation, young monkeys did not display beta-amyloid-immunoreactive profiles within any CNS structure regardless of treatment. Qualitative observations revealed that aged monkeys displayed numerous beta-amyloid plaque like structures within the amygdala and hippocampus as well as limbic and neocortices. The amount of beta-amyloid immunoreactivity (beta-amyloid load) was quantified bilaterally within the temporal neocortex of these animals. The beta amyloid load within the temporal neocortex of aged monkeys was highly variable but did not differ across treatment groups. These data indicate that chronic short-term administration of NGF does not affect the expression of beta-amyloid in the young or the aged primate brain. PMID- 9217096 TI - An acid-treatment method for the enhanced detection of GDNF in biological samples. AB - Glial cell line-derived neurotrophic factor (GDNF), a distant member of the transforming growth factor-beta (TGFbeta) family, is a protein that is essential for the survival of dopaminergic, motor, and peripheral neurons. To facilitate its study, we and others have developed sensitive (low pg/ml) enzyme-linked immunosorbant assays (ELISA) to quantitate endogenous concentrations of GDNF, along with neurotrophin-3 (NT-3) and nerve growth factor (NGF). However, endogenous tissue levels of GDNF in adult animals are not readily detected by ELISA and do not correlate well with message RNA. Based upon previously described methods for the extraction of TGFbeta from tissue samples, we have developed an acid-treatment procedure to allow the quantification of total endogenous GDNF. This procedure also was evaluated for use when measuring total endogenous levels of NT-3 and NGF from biological samples. The acid-treatment procedure increases the detectable amounts of GDNF, NT-3, and NGF in all tissue samples and most of the serum samples tested. Moreover, these values were as much as 35 times greater than those detected using traditional extraction buffers. Such elevated concentrations likely resulted from the acid treatment promoting the dissociation of ligands from receptors or binding proteins, thereby making more of the analyte available to be measured in the ELISA. These findings indicate that appropriate sample treatment is essential for the measurement of total endogenous neurotrophic factors. PMID- 9217097 TI - Driving predictive modelling on a risk assessment path for enhanced food safety. AB - How do we best protect our citizens to allow the highest quality of life? Where do we put our food safety resources so that we gain the greatest positive impact? Risk assessment provides the critical scientific basis for these types of important risk management decisions. Increasingly, risk assessment is used to guide legislated and voluntary changes intended to improve safety, yet its formal application for enhanced food safety is in its infancy. Risk assessment includes disease characterization. dose-response assessment, exposure assessment, and risk characterization. Quantitative data is critical for risk assessment to realize its full value, yet much of our knowledge about the incidence of pathogens or toxins in foods, dose-response knowledge, incidence of acute food-borne illness, incidence of chronic sequelae, and cost of food-borne illness is qualitative or estimates are controversial. Predictive modelling should help to improve estimates and thereby allow quantitation of food safety risks. Predictive modelling will also find application for assessing prevention strategies in risk management. PMID- 9217098 TI - Use of predictive microbiology in microbial food safety risk assessment. AB - Microbial risk assessment is a newly emerging discipline in the area of food safety. One of the difficulties associated with microbial risk assessment is in determining the number of microorganisms in food at a given time, i.e.. estimating exposure of an individual to the microorganism. Numbers of bacteria in food can change at all stages of food production and processing, depending on the nature of the food and the way it is handled, stored and processed. Predictive microbiology can be used to estimate changes in bacterial numbers, allowing exposure of an individual to a pathogen to be assessed. A survey was sent to scientists in the food industry to determine their perspective on the role of predictive microbiology in conducting microbial risk assessments. In this paper, responses to that survey are presented, as well as examples of the potential risk of foodborne illness from a cooked meat product contaminated with Staphylococcus aureus and hamburger contaminated with Salmonella. PMID- 9217099 TI - Use of predictive microbiology in meat hygiene regulatory activity. AB - New Zealand is a supplier of refrigerated raw meat to world markets. To maintain this supply, from regulatory and commercial perspectives, production standards need to deliver products that are both hygienically adequate and commercially viable. A dynamic Temperature Function Integration (TFI) model, as a form of predictive microbiology, was used jointly by regulators and processors to develop justifiable criteria for the management of refrigeration during the production of hot and warm-boned meat, the post-slaughter handling of ovine carcasses and the handling of offals. Current processes operating according to accepted standards for Good Manufacturing Practice (GMP) were quantified in terms of TFI. The hygienic adequacy of new processes were similarly determined using the TFI model and compared to relevant GMP standards. From a regulatory perspective, the dynamic TFI model has provided a rapid and cost effective method of quantifying a temperature dependent process in terms of the potential for microbial proliferation. It has also produced a method for determining parameters for new or intended processes by comparing the potential for microbial proliferation with previously validated outputs, and has complemented traditional quantitative microbiology to provide a rapid, cost effective method of verifying that a process is performing according to design parameters. However, it could not be used to validate standards for processing in the absence of existing standards for GMP, or in the absence of microbial standards previously established using the principles of risk assessment. PMID- 9217100 TI - Development of a quantitative risk assessment model for Salmonella enteritidis in pasteurized liquid eggs. AB - The performance of hazard analyses and the establishment of critical limits by the food industry are both hampered by the inability to directly relate food processing operations from farm-to-table with their public health impact. Using a 'unit operations' and stochastic simulation approach, data on the frequency of pathogens in raw ingredients, predictive microbiology models for growth and inactivation (thermal and non-thermal), and dose-response models for infectivity were integrated to create a quantitative risk assessment model for a Salmonella enteritidis infection from thermally processed liquid whole eggs made into mayonnaise in the home. The risk assessment indicated pasteurization provides sufficient consumer protection from a high incidence of infected birds and from temperature abuse between the farm and the egg breakers. However scenarios showed how inadequate pasteurization temperatures and/or temperature abuse during storage leads to a hazardous product. This dynamic approach to modeling risk should aid in identification and setting critical control points and assessing the impact of altering food formulations or processes. PMID- 9217101 TI - A regulatory perspective on the potential uses of microbial risk assessment in international trade. AB - The recent ratification of the World Trade Organisation Agreement will arguably be the most important factor in developing new sanitary measures for the international trade in food over the next decade. There is a markedly increased desire for quantitative data on the microbial risks associated with different classes of foods, and traditional good manufacturing practice (GMP)-based food hygiene requirements are coming under increasing challenge. As the risk assessment paradigm is increasing applied and as decision-making criteria for risk management become established, more emphasis will be placed on predictive microbiology as a means of generating exposure data and establishing critical limits for Hazard Analysis Critical Control Point (HACCP) plans. In this respect, developing international guidelines for risk management arguably presents the greatest challenge in establishing and maintaining quantitative Sanitary and Phytosanitary (SP) measures for food in international trade, and for judging their equivalence. Where specific industry sectors and regulators do not have jurisdiction over the entire food chain, from production of raw materials through to consumption, it will be difficult to apply the risk assessment paradigm in the design of HACCP plans. Thus, it appears that default to food safety objectives for many segments of food production chains subject to application of HACCP plans is inevitable in the medium term. PMID- 9217102 TI - MPN-PCR-quantification method for staphylococcal enterotoxin c1 gene from fresh cheese. AB - PCR detection methods have been extensively used in diagnostic microbiology. However, a lack of a simple and reliable method for quantification of the PCR products has partly hindered the use of PCR in routine food laboratories. The quantification of PCR products can be done by combining the principles of MPN statistics and PCR technique. We have developed a simple and sensitive MPN-PCR assay for detection and enumeration of enterotoxin C producing Staphylococcus aureus NCTC 10655 from fresh cheese. By amplifying single copy chromosomal enterotoxin C gene fragment, we could detect as little as 20 cfu/g. By Moran's test, most of the DNA dilution series appeared to fulfill the basic mathematical assumptions of ordinary MPN methods. The analysis with MPN-PCR took one day to perform compared with three days analysis time with plate counting. This MPN-PCR method can be readily applied with different primer systems without extensive development work. PMID- 9217103 TI - Weak acid inhibition of fermentation by Zygosaccharomyces bailii and Saccharomyces cerevisiae. AB - The inhibition kinetics of fermentation by Zygosaccharomyces bailii and Saccharomyces cerevisiae were evaluated for weak carboxylic acids. Several regression equations were tried to fit the experimental data, most cases being best fitted to exponential curves. The following parameters were determined: i) acid concentration responsible for 50% inhibition of fermentation (C50%); ii) area under the regression curve up to that concentration (A50%) and iii) exponential inhibition constant (k(i)). These parameters were compared according to their ability to express the inhibitory effect of each acid. In broad terms, the values of k(i) in association with minimum inhibitory concentrations (x(min)), were found best to express the inhibitory effect of the weak acids. However, C50% values were satisfactorily correlated with k(i). The value of A50% more precisely reflected the occasional stimulatory effect of low concentrations of weak acids. Comparison of inhibition parameters for Z. bailii and for S. cerevisiae revealed a higher resistance of the former to acetic, propionic, butyric and benzoic acids and similar resistance to caproic, caprylic and sorbic acids. Previous cultivation in the presence of acetic, propionic and benzoic acids showed a distinct influence on the resistance of both yeasts, although it did not always induce cellular adaptation. Fermentation inhibition showed a good correlation with the lipid solubility of weak acids suggesting that the acids interact with the hydrophobic regions of cell membranes. PMID- 9217104 TI - Antilisterial activity of three bacteriocins used at sub minimal inhibitory concentrations and cross-resistance of the survivors. AB - The kinetics of inhibition of Listeria innocua strain Lin11 in a nutrient broth containing either nisin, pediocin AcH, or enterococcin EFS2 at concentrations ensuring a 2-3 log reduction of the population were first investigated. The rate of inhibition differed considerably between the bacteriocins. At the time that maximum viability loss occurred in the cultures containing either nisin (12 AU ml(-1)) or pediocin AcH (50 AU ml(-1)) the survivors resumed growth, although the medium retained most of its initial inhibitory activity. The survivors displayed increased resistance not only toward the bacteriocin they were in contact with, but toward the two other bacteriocins under study. PMID- 9217105 TI - Extremely halotolerant bacteria characteristic of fully cured and dried cod. AB - Gram-positive cocci were isolated in high numbers from salted codfish during processing. They were found to be the main bacterial type in fully cured and dried salted cod. Phenotypic characterization of 37 strains showed them to belong to the novobiocin resistant staphylococci, most likely Staphylococcus arlettae or xylosus. Based on sequencing of 16S rDNA and comparison of 700 bases it was concluded that they should be assigned to the species Staphylococcus arlettae. They were found to be extremely halotolerant, growing well at salt concentrations from 0.06 M NaCl, and even displaying clear growth at 4.5 M NaCl. Likewise, the strains grew over a wide temperature range, from 8 to 45 degrees C. Optimal growth conditions were found to be at 0.4-0.6 M NaCl and 30-32 degrees C. This is all in accordance with findings for related staphylococci that have been isolated from other heavily salted meat or fish products. PMID- 9217106 TI - Assessment of the hygienic performances of hamburger patty production processes. AB - The hygienic conditions of the hamburger patties collected from three patty manufacturing plants and six retail outlets were examined. At each manufacturing plant a sample from newly formed, chilled patties and one from frozen patties were collected from each of 25 batches of patties selected at random. At three, two or one retail outlet, respectively, 25 samples from frozen, chilled or both frozen and chilled patties were collected at random. Each sample consisted of 30 g of meat obtained from five or six patties. Total aerobic, coliform and Escherichia coli counts per gram were enumerated for each sample. The mean log (x) and standard deviation (s) were calculated for the log10 values for each set of 25 counts, on the assumption that the distribution of counts approximated the log normal. A value for the log10 of the arithmetic mean (log A) was calculated for each set from the values of x and s. A chi2 statistic was calculated for each set as a test of the assumption of the log normal distribution. The chi2 statistic was calculable for 32 of the 39 sets. Four of the sets gave chi2 values indicative of gross deviation from log normality. On inspection of those sets, distributions obviously differing from the log normal were apparent in two. Log A values for total, coliform and E. coli counts for chilled patties from manufacturing plants ranged from 4.4 to 5.1, 1.7 to 2.3 and 0.9 to 1.5, respectively. Log A values for frozen patties from manufacturing plants were between < 0.1 and 0.5 log10 units less than the equivalent values for chilled patties. Log A values for total, coliform and E. coli counts for frozen patties on retail sale ranged from 3.8 to 8.5, < 0.5 to 3.6 and < 0 to 1.9, respectively. The equivalent ranges for chilled patties on retail sale were 4.8 to 8.5, 1.8 to 3.7 and 1.4 to 2.7, respectively. The findings indicate that the general hygienic condition of hamburgers patties could be improved by their being manufactured from only manufacturing beef of superior hygienic quality, and by the better management of chilled patties at retail outlets. PMID- 9217107 TI - A heterogeneous population model for the analysis of bacterial growth kinetics. AB - A two-compartment, heterogeneous population model (HPM) was derived using the simulation software SB ModelMaker to describe the growth of Listeria monocytogenes in bacteriological media at 5-35 degrees C. The model assumed that, at time t = 0, the inoculum was distributed between two distinct compartments, Non-Growing and Growing, and that growth could be described by four parameters: initial total cell population (N0), final maximum cell population (Nmax), maximum specific growth rate (mu(max)), and initial cell population in the Growing compartment (G0). The model was fitted to the data by optimizing the four parameters, and lag phase duration (lambda) was calculated. The resulting values of mu(max) and lambda were similar to those determined using the modified Gompertz equation. A new parameter, w0, was defined which relates to the proportion of the initial cell population capable of growth, and is a measure of the initial physiological state of the cells. A modified model in which mu(max) was replaced with a temperature function, and w0 replaced G0, was used to predict the effect of temperature on the growth of L. monocytogenes. The results of this study raise questions concerning the current definition of the lag phase. PMID- 9217108 TI - Water availability and the survival of Salmonella typhimurium in porous systems. AB - The survival of Salmonella Typhimurium LT2 in randomly packed beds of glass beads, microporous silica particles and Sephadex microspheres is examined. It is shown that the decrease in the percentage cell recovery in these porous materials at reduced water content is not correlated with the global water activity as determined by conventional vapour pressure measurements but rather with the osmotic shock induced by the sudden redistribution of water and air among the microscopic pores in the matrix surrounding the cells. For this reason the bacterial survival and growth data correlates best with physical measurements, such as NMR and electrical conductivity, which are sensitive to the microscopic air-water distribution. The implications of this observation in food safety and preservation are discussed. PMID- 9217109 TI - Identification and quantification of risk factors regarding Salmonella spp. on pork carcasses. AB - The main elements of a descriptive epidemiological model for Salmonella spp. in Dutch pig slaughterlines, and the subsequent quantification of risk factors regarding the contamination of carcasses, are described. There is a strong correlation between the number of live animals that carry Salmonella spp. in their faeces and the number of contaminated carcasses at the end of the slaughterline. Live animals that carry Salmonella spp. are 3-4 times more likely to end up as a positive carcass than Salmonella-free animals. Currently, about 70% of all carcass contamination results from the animals themselves being carriers, and 30% because other animals were carriers (i.e. cross contamination). Furthermore, it is estimated that in general between 5-30% of the carcasses produced may contain Salmonella spp. With respect to carcass contamination with Enterobacteriaceae and Salmonella spp., inadequately cleaned polishing machines (odds ratio, OR, 6) and 'inapt procedures during evisceration' (OR 11), i.e. faulty evisceration and hygiene practices, are the most important risk factors. An estimated 5-15% of all carcass contamination with Salmonella spp. occurs during polishing after singeing. The remainder is the result of current evisceration practices (55-90%) and, to a lesser extent, further processing (5 35%), i.e dressing, splitting and meat inspection. Less likely Salmonella spp. already present on the skin of the live animals survive scalding and singeing. However, because pigs are the only important source for the Salmonella contamination of the line and the carcasses produced, it can also be concluded that if Salmonella-free pigs were produced, consumers could be provided with virtually Salmonella-free pork. As long as Salmonella-positive animals enter abattoirs, there will always be transmission of Salmonella spp. to consumers, even if the process is carried out according to stringent codes of good manufacturing practices (GMP). EU regulations should, therefore, allow for the decontamination of caracasses with a safe substance, e.g. lactic acid, on the condition that the slaughterhouse strictly adhers to GMP principles. PMID- 9217110 TI - Hydrolysis of esters by staphylococci. AB - The objective of this work was to characterize the hydrolysis of esters by staphylococci in order to understand if they could contribute to the release of free fatty acids in sausage. Cell-free extracts and extracellular concentrates of staphylococci were examined for esterase activities against p-nitrophenyl esters and for lipolytic activities against triolein. Staphylococci showed intracellular and extracellular esterase activities with different esterase electrophoretic patterns. Cell-free extracts of S. xylosus, S. warneri and S. saprophyticus preferentially hydrolysed p-nitrophenyl butyrate whereas their extracellular concentrates were mainly active against p-nitrophenyl butyrate, p-nitrophenyl caproate and p-nitrophenyl caprylate. In addition their extracellular concentrates hydrolysed triolein. The two strains of S. carnosus differed as they did not show pronounced p-nitrophenyl substrate specificity and did not hydrolyse triolein. Staphylococci hydrolysed esters at a high rate between 15 and 25 degrees C; acidic conditions inhibited the hydrolysis. The hydrolysis was also reduced when the water activity was decreased by addition of polyethylene glycol or glycerol. PMID- 9217111 TI - Detection of aflatoxinogenic fungi in figs by a PCR reaction. AB - A PCR reaction was used to detect aflatoxinogenic Aspergillus flavus strains in contaminated figs. The reaction records the presence of three aflatoxin biosynthesis genes, namely the norsolorinic acid reductase (nor-1), versicolorin A dehydrogenase (ver-1) and sterigmatocystin-o-methyltransferase: (omt-A). The reaction gave a triplet pattern in the presence of DNA from A. flavus isolated from pure cultures. The reaction gave the same PCR products when pure fungal DNA was mixed with pure DNA isolated from figs, but the sensitivity was reduced by a factor of 10. The same set of bands was observed when isolated DNA from infected figs was used as template DNA but no signal was visible when DNA from uninfected figs was used as template. PMID- 9217112 TI - Effect of temperature and agitation on enrichment of Escherichia coli O157:H7 in ground beef using modified EC broth with novobiocin. AB - The effects of temperature and agitation on the enrichment of Escherichia coli O157:H7 in meat using modified EC broth with novobiocin (mEC + n) were studied. Enrichment at 37 degrees C was compared to 42 degrees C, both with and without shaking. Incubation at 42 degrees C without shaking effectively suppressed ground beef microflora while allowing good growth of E. coli O157:H7 cells. Cells inoculated into ground meats (beef, pork, turkey) were readily detected by enrichment for 24 h in mEC + n at 42 degrees C without shaking, followed by screening the enrichment cultures using a rapid and inexpensive commercially available enzyme immunoassay system, the E. coli O157 Rapitest. PMID- 9217114 TI - Influence of aging treatment on the bacterial quality of South African springbok (Antidorcas marsupialis marsupialis) wholesale cuts. AB - The left sides of 20 springbok carcasses were aged with the skin on (treatment 1), while the right sides were aged without the skin (treatment 2). Another 20 carcasses were skinned, halved and cut into wholesale cuts. The loin and leg cuts from the right sides were deboned before vacuum packaging (treatment 3), while the loin and leg cuts from the left sides were vacuum packed with the bone in (treatment 4). All the carcass sides (36 h post mortem) and vacuum packed cuts (36 h post mortem) were aged for either 2, 5, 12 and 19 days respectively (ca. 0 degrees C). The examined groups of bacteria indicate that, for an ageing period of 12 days or less, vacuum packaging does not have an advantage over the hung in air method. However, when considering an extended ageing period ( > 12 days) vacuum packaging ensures that spoilage bacteria are inhibited and that the Enterobacteriaceae group of bacteria do not increase (ca. 0 degrees C), while the results clearly show that this is not the case with the hung in air ageing treatments (1 and 2). PMID- 9217113 TI - Toxinogenicity of heat-resistant fungi detected by a bio-assay. AB - The ciliostatic activity of exo- and endometabolites of 125 (100%) heat-resistant fungal isolates was evaluated in vitro by a bioassay on tracheal tissue cultures of 1 day old chicks. For 11 (9%) of the isolates investigated chloroform extractable secondary metabolites were detected in the medium. For 29 (23%) they were detected in mycelium and spores and for 25 (20%) they were detected in the medium, mycelium and spores at the same time. Especially Dichotomomyces cejpii and Eupenicillium baarnense strains displayed ciliostatic activity. Extracts of Gilmaniella humicola, Talaromyces avellaneus and Talaromyces bacillisporus isolates were not found ciliostatic in our experiment. The chloroform-extractable metabolites of Neosartorya fischeri ATCC 96469 and T. avellaneus ATCC 96465 strains had no ciliostatic activity, either, but G. humicola ATCC 96467 produced ciliostatic metabolites detected in the medium, the mycelia and the spores. PMID- 9217115 TI - A modified agar medium for the screening of proteolytic activity of starter cultures for meat fermentation purposes. AB - An agar medium, used in the screening of proteolytic activity of dairy-related bacteria, was adapted for assessing the proteolytic capacity of bacteria which were of possible use in meat fermentations. Freeze dried myofibrils, extracted from pork muscle, were incorporated in the medium. The agar plates were inoculated with 20 microl of overnight cultures of different starter strains, and incubated at 30 degrees C for 48 h. After incubation, proteolytic bacteria produced clear zones. Coomassie brilliant blue stain was employed to facilitate the detection of these zones. Proteolytic activity was confirmed in an enzymatic test. PMID- 9217116 TI - Ecology of inoculated and spontaneous fermentations in Rioja (Spain) musts, examined by mitochondrial DNA restriction analysis. AB - Analysis of mitochondrial DNA restriction patterns was used to study the introduction of a selected strain of Saccharomyces cerevisiae for fermentation of non-sterile musts of La Rioja (Spain). All of the isolates from the inoculated musts showed the restriction pattern of the selected strain. The same technique was used to study the spontaneous fermentation of musts, showing that a few strains were responsible for the fermentations. One of the strains identified from the spontaneous fermentations had been identified in a previous vintage. PMID- 9217117 TI - Tracheotomy and the intensive care unit patient. AB - Transportation of the intensive care unit (ICU) patient to the operating room for tracheotomy has been implicated as an unnecessary source of complications and has been cited as a relative indication for percutaneous tracheotomy. However, there is very little evidence in the literature to support this claim. We evaluated 100 consecutive patients who were transported from the ICU to the operating room for tracheotomy. There were no complications related to patient transportation. A total of five complications occurred, all unrelated to patient transportation. Two patients receiving pressure control ventilation developed a pneumothorax on postoperative days 7 and 8, respectively. There were three minor complications directly related to the tracheotomy: peristomal cellulitis, tracheitis, and hemorrhage of less than 25 cc on postoperative day 1. The minor complications were treated appropriately and resolved without any adverse sequelae. We provide a detailed review of 100 consecutive ICU patient tracheotomy cases and compare this with 109 tracheotomies in non-ICU patients. Transportation of the ICU patient does not appear to increase the risk of complications during tracheotomy and should not be cited as a cause of complications in the percutaneous tracheotomy literature. The results with standard surgical tracheotomy in the controlled setting of the operating room should serve as the standard by which other procedures are judged. PMID- 9217118 TI - Electrocautery versus carbon dioxide laser for uvulopalatoplasty in the treatment of snoring. AB - Laser-assisted uvulopalatoplasty is a popular method for reducing snoring. Drawbacks are the large initial expense of the laser unit and related equipment and required safety precautions. The equipment required for electrocautery for cautery-assisted uvulopalatoplasty is significantly less expensive to obtain and operate compared with the carbon dioxide laser. Ninety-eight patients were randomly assigned to one of two treatment groups to undergo uvulopalatoplasty: one performed with the carbon dioxide laser and the other with electrocautery. We compared postoperative pain, time off work, efficacy, and the number of treatments required to achieve a satisfactory result. We found no statistically significant difference in any of these parameters between the two treatment groups (P > 0.05). Our data show that the use of the carbon dioxide laser offers no advantage over electrocautery in performing uvulopalatoplasty to treat snoring. PMID- 9217119 TI - Rhinocerebral mucormycosis: evolution of the disease and treatment options. AB - Rhinocerebral mucormycosis is recognized as a potentially aggressive and commonly fatal fungal infection. The classic presentation is involvement of nasal mucosa with invasion of the paranasal sinuses and orbit. Mucormycosis is most commonly seen in association with diabetic ketoacidosis, but disease demographics have changed with the onset of AIDS and the advent of powerful immunosuppressive drugs. Treatment includes aggressive debridement, systemic antifungal therapy, and control of underlying comorbid factors. Although surgical intervention remains essential, advances in medical therapy have permitted a more limited surgical approach to minimize functional loss without compromising survival. We present the UCLA experience with rhinocerebral mucormycosis from 1955 to 1995, with emphasis on the evolution of disease presentation and alternative treatment options. PMID- 9217120 TI - Intracranial complications of sinusitis. AB - Intracranial suppurative complications of sinusitis remain a challenging and contemporary topic. To determine the prevalence of sinogenic sources in intracranial infectious complications, we reviewed the records at a large public hospital between 1985 and 1995. There were 203 patients with 212 suppurative intracranial infections. Sinogenic sources were identified in 12 patients with 19 infections. Most patients had ethmoid or frontal sinusitis. We discuss the presentation, microbiology, diagnosis, treatment, and clinical course of these 12 cases. The diagnosis of intracranial complications of sinusitis requires a high index of suspicion and radiographic imaging of the head and paranasal sinuses. The mean hospital stay was 31.4 days and all 12 patients survived, although three patients had significant neurologic sequelae. PMID- 9217121 TI - Suprastomal granulation tissue and pediatric tracheotomy decannulation. AB - Although numerous decannulation techniques have been reported, often involving costly sleep studies, repetitive laser procedures, and tracheotomy tube "downsizing," no established standard of care exists. We advocate the following simple, minimally invasive decannulation protocol. After excluding concomitant airway lesions, suprastomal granulation is removed transtomally by an endoscopically guided rongeur. A tracheotomy tube is then fashioned with a fenestration centered in the tracheal lumen. Decannulation occurs if the patient maintains adequate ventilation over a 12- to 24-hour observation period with the fenestrated tracheotomy capped. Over 18 months we prospectively followed 10 consecutive children presenting as potential decannulation candidates. Using the aforementioned technique, nine of 10 patients were successfully decannulated (average follow-up, 11.5 months). The postoperative capped fenestrated tracheotomy trial provides a realistic assessment of preparedness for decannulation. We recommend this protocol as a rapid, efficient, and cost effective means of achieving decannulation. PMID- 9217122 TI - Efficacy of tympanomastoid surgery for control of infection in active chronic otitis media. AB - The efficacy of surgery in controlling infection in 272 tympanomastoidectomy procedures for chronic otitis media (COM) was assessed by means of a four-point rating scale that incorporated both symptoms and signs, such as the presence or absence of otorrhea and granulation tissue. Of the 272 procedures, 170 were performed for COM with cholesteatoma and 102 were for active COM with granulation tissue but no cholesteatoma. Forty-seven percent were primary procedures, and 53% were revisions. Minimum follow-up was 12 months for all cases, with a mean of 30 months. Adequate control of infection occurred in 248 (91%) of the 272 cases. Of the 24 cases (9%) that developed persistent infection, 10 were controlled with a combination of oral and topical antibiotics and/or delayed skin grafting in the office. Thus overall satisfactory control of infection was achieved in 258 of 272 cases (95%). The outcome was influenced by the diagnostic category of COM: COM with cholesteatoma did significantly better than COM with granulation tissue (P = 0.02). The outcome was not influenced by the following variables: primary versus revision surgery, canal wall-up versus canal wall-down surgery, and extent of disease. The results suggest that active COM with granulation tissue may be more difficult to control than COM with cholesteatoma. PMID- 9217123 TI - Can sensitized lymphocytes retain reactivity to inner ear antigens after retrieval from frozen storage? AB - Immune inner ear disease results in rapidly progressive, bilateral sensorineural hearing loss and is one of the few forms of sensorineural hearing loss that can be treated medically. The purpose of this study is to identify and preserve several populations of sensitized lymphocytes from patients with immune inner ear disease as a first step toward cloning autoreactive T cells, in order to study the pathogenesis of disease. Lymphocytes from four patients with high reactivity (stimulation index of 2.5 or greater) were placed in frozen storage. At 8 to 14 months they were thawed and restimulated. All four samples were viable. Two reacted again to inner ear homogenate, but with different intensities. Some lymphocytes sensitized to inner ear antigens can retain reactivity after frozen storage. This methodology may be useful to clone highly reactive T cells. PMID- 9217124 TI - Periauricular cysts and sinuses. AB - Periauricular cysts, sinuses, and fistulas occur commonly in the pediatric population. They arise from developmental defects of the first branchial cleft and first branchial arch. In most instances the diagnosis and management of these conditions are straightforward, but exceptional presentations sometimes occur. Failure to recognize these unusual cases may result in inadequate treatment and subsequent recurrence, and even if the correct diagnosis is made, surgical management of these lesions may be complicated. A series of 15 cases of periauricular congenital lesions is reviewed, of which three cases illustrating a diagnostic or surgical challenge are presented. The embryology, presentation, and management of these anomalies are discussed. This is one of the largest series of first branchial cleft anomalies reported in the literature, and our paper uniquely discusses first branchial cleft anomalies and preauricular sinuses together, with an emphasis on the surgical management of facial nerve, external ear, and middle ear involvement. PMID- 9217126 TI - Midline mandibular osteotomy: an analysis of functional outcomes. AB - Although the oncologic validity and perioperative complications of midline mandibular osteotomy are well described, little attention has been directed toward the long-term functional problems that may be associated with its use. Thirty-one patients who had undergone this procedure were examined to assess postoperative sensation, temporomandibular joint (TMJ) function, occlusion, and cosmesis. The majority (27 of 31) patients had some sequelae but these were minor in nature. Twenty of 31 patients had abnormal sensation, 24 of 31 noted a changed occlusion, and 15 of 31 had signs or symptoms of TMJ myofascial pain. Although patients should be advised of the potential for functional problems with this procedure, they can be reassured that these are likely to be relatively minor in significance. If technically feasible and if an exact restoration of occlusion is a priority, a prefabricated lingual splint should be used. PMID- 9217125 TI - Factors affecting smoking cessation in patients with head and neck cancer. AB - The role of tobacco in the etiology of upper aerodigestive tract carcinomas is well established. Smoking decreases the effectiveness of cancer therapy and increases the risk of all treatment modalities. Smoking adversely affects the general health of the cancer survivor and places the patient at risk of developing additional primary tumors. The smoking habits of head and neck cancer patients were evaluated using a questionnaire administered at two tertiary head and neck cancer centers. Demographic factors, level of exposure, tumor stage and location, treatment modalities, concomitant alcohol use, and cessation methods were examined. Results demonstrate a high rate of smoking cessation at the time of cancer diagnosis. Significant demographic factors were not identified. Physical barriers to continued smoking because of cancer treatment as well as counseling at the time of tumor diagnosis were the most effective deterrents to continued tobacco use. Heavy alcohol use was a negative predictor of smoking cessation. Pharmacologic aids alone were found to be of no value. This study demonstrates the difficulties with smoking cessation in head and neck cancer patients, and emphasizes the importance of intervention by the otolaryngologist head and neck surgeon. PMID- 9217127 TI - Ingestion of caustic hair relaxer: is endoscopy necessary? AB - Hair relaxer, a commercially available alkaline product, is commonly the offending agent in caustic ingestion. These patients often experience oral cavity and facial burns; however, no clinically significant esophageal injuries have been reported. Therefore, we questioned the therapeutic and economic efficacy of the "standard treatment protocol" that includes hospitalization and endoscopic evaluation. Twenty-six patients over a 7-year period presented to our institution having ingested hair relaxer. Presenting signs and symptoms, esophageal findings, and cost of the standard treatment protocol were reviewed. Also, we analyzed the caustic potential and current packaging of hair relaxer. Our findings support modifications in the standard treatment protocol for hair relaxer ingestion including elimination of hospitalization and endoscopy in most patients. We also question compliance with childproof packaging laws and suggest avenues for prevention of hair relaxer ingestion. PMID- 9217128 TI - Hyperparathyroidism associated with a chronic hypothyroid state. AB - Reports of the coexistence of hyperparathyroidism and thyroid disease have raised the issue of a possible etiologic relationship. The present study tests the hypothesis that chronic elevation of thyroid-stimulating hormone (TSH) is related to the development of hyperparathyroidism. Four groups of 60 female rats were treated as follows: group 1, control; group 2, propylthiouracil (PTU) 0.0025%; group 3, PTU 0.0025% plus thyroxine, 5 microg two times per week; and group 4, only thyroxine. The animals' serum calcium, phosphorus, TSH, thyroxine, and parathyroid hormone (PTH) levels were evaluated at 0, 6, 12, and 18 months. Significant elevation of TSH was sustained throughout the 18 months in groups 2 and 3. The PTH levels were also significantly elevated in both group 2 and group 3 animals (P = 0.02). The histopathologic features of the parathyroids were evaluated at 18 months. In the group 2 (PTU only) animals, which had profound hypothyroid, 44% developed parathyroid adenomas. In the group 3 (PTU plus thyroxine) animals, who had mildly elevated TSH levels, 53% developed parathyroid adenomas. These findings are consistent with the hypothesis that prolonged TSH stimulation may lead to hyperparathyroidism in the rat model. PMID- 9217129 TI - Congenital encephalocele of the medial skull base. AB - Meningoencephaloceles of the temporal bone are rare. Although most often seen following otologic surgery or trauma, congenital meningoencephaloceles can exist. The clinical presentation, diagnostic evaluation, and surgical management of three patients with congenital meningoencephalocele are presented. Two of the three patients presented to our institution with recurrent episodes of meningitis; one presented with partial complex seizures. Diagnostic evaluation included temporal bone computed tomography with magnetic resonance imaging. In two patients, defects were imaged following high-pressure subarachnoid cisternography with computed tomography. All three patients were found to have congenital defects in the area of Meckel's cave. Early recognition of congenital meningoencephalocele is important to avoid delay of definitive surgical management and neurologic sequelae. PMID- 9217131 TI - Stapedectomy in patients with small air-bone gaps. AB - Controversy exists concerning stapedectomy for patients with small air-bone gaps. The purpose of this study was to examine the results for patients who had a stapedectomy to correct a small (10 dB or less) air-bone gap. One hundred fifty four patients with suspected otosclerosis were explored and a stapedectomy was performed in 136 (88.3%) of these cases. The mean pure-tone average (PTA) improved 16.7 dB and overdosed the preoperative bone conduction PTA by 8.1 dB. The majority of the stapedectomy patients (89.7%) had a PTA closure greater than or equal to 0 dB. These results showed that stapedectomy can be an effective procedure for eliminating and overdosing even small air-bone gaps due to otosclerosis. PMID- 9217130 TI - Pediatric respiratory papillomatosis: prognostic role of viral typing and cofactors. AB - Children with recurrent respiratory papillomatosis vary greatly in their clinical disease course. Many have mild disease with eventual remission while others present with an early aggressive airway obstructive course. This study consisted of 24 pediatric patients whose specimens underwent polymerase chain reaction analysis for cytomegalovirus (CMV), herpes simplex virus (HSV), and human papillomavirus (HPV) type. Nineteen of 24 specimens contained enough DNA for this study. None of the specimens were found to contain DNA from HPV-16, -18, -31, 33; CMV; or HSV, which contrasts with our previous findings in adults. Ten patients were infected by HPV-11 and seven of these underwent tracheotomy because of an aggressive tumorigenic clinical course. Nine patients were infected by HPV 6 alone of whom only two required a tracheotomy (P = 0.05, Fisher's Exact Test). The early airway obstructive course associated with HPV-11, however, had no bearing on achieving eventual disease remission, with decannulation achieved in eight of nine children. PMID- 9217132 TI - Adenovirus and respiratory syncytial virus in chronic sinusitis using polymerase chain reaction. AB - The aim of this study is to investigate the role of adenovirus and respiratory syncytial virus (RSV) in chronic sinusitis using the polymerase chain reaction (PCR) to assay for the viruses. PCR has proved to be more sensitive and specific than viral cultures and immunoassays in the detection of viruses. Adenovirus and RSV are among the most common viruses to cause upper respiratory tract infections. Sinus mucosa biopsies from 20 patients undergoing endoscopic sinus surgery were sterilely collected. Four specimens (20%) tested positive for RSV by PCR and none tested positive for adenovirus. Only one specimen tested positive for RSV and one for adenovirus by viral culture and immunofluorescence. Bacterial cultures tested positive in 40% of the 20 specimens. PCR can be used to detect RSV in patients with chronic sinusitis and PCR is more sensitive than viral culture and immunofluorescence techniques on sinus polyps and mucosa. PMID- 9217133 TI - Myofibroblast accumulation induced by transforming growth factor-beta is involved in the pathogenesis of nasal polyps. AB - Myofibroblasts that express alpha-smooth muscle actin (alpha-SMA) are detected in many chronic inflammatory diseases. Transforming growth factor-beta (TGF-beta) is a potent inducer of myofibroblast accumulation in tissues. In this study, scattered myofibroblasts and TGF-beta were quantified and localized in nasal polyps (NPs) and normal nasal mucosa (NM). NPs were sampled in 16 patients during ethmoidectomy and NM was obtained from 10 control subjects during rhinoplasty. alpha-SMA and TGF-beta were detected using immunohistochemistry and the numbers of labeled cells were quantified (alpha-SMA and TGF-beta indices) and compared between NPs and NM. In eight NPs, in which the pedicle was preserved, alpha-SMA and TGF-beta were evaluated and compared in the pedicle, central, and tip areas. Finally, TGF-beta expression was compared between low (zone 1), moderate (zone 2), and high (zone 3) zones of alpha-SMA positivity. alpha-SMA and TGF-beta indices were significantly higher in NPs than in NM. In the eight selected NPs, alpha-SMA-positive cells were significantly more abundant in the pedicle than in the central and tip areas, whereas TGF-beta-positive cells were significantly more numerous in the pedicle than in the tip area. The number of TGF-beta positive cells was significantly higher in zone 3 than in zone 1 of alpha-SMA positivity. Myofibroblasts, which are abundant in NPs but rare in NM, could be involved in the growth of NPs by inducing extracellular matrix accumulation. The local development of myofibroblasts in NPs could be controlled by TGF-beta, locally produced by inflammatory cells. PMID- 9217134 TI - Soluble intercellular adhesion molecule-1 level in sera is elevated in perennial allergic rhinitis. AB - Soluble intercellular adhesion molecule-1 (sICAM-1) in sera was measured in some allergic disorders, but serum sICAM-1 levels in perennial allergic rhinitis remain to be determined. Our study was aimed at elucidating whether the serum sICAM-1 levels in patients with perennial allergic rhinitis are different from those in nonatopic healthy volunteers and whether immunotherapy can modulate sICAM-1 levels. Serum sICAM-1 was determined in 20 nonallergic volunteers and 137 patients with perennial allergic rhinitis by a sandwich enzyme-linked immunosorbent assay. Our study demonstrated that the level of sICAM-1 in untreated patients is significantly elevated, as compared with nonatopic subjects. Immunotherapy could decrease sICAM-1 in perennial allergic rhinitis, but this suppressive effect became apparent only after many years of immunotherapy. In patients on immunotherapy, a close correlation was observed between sICAM-1 and nasal symptom scores. To take these lines of evidence together, a decrease in sICAM-1 might be related to the working mechanism of immunotherapy, and serum sICAM-1 could be used to monitor the effect of immunotherapy. PMID- 9217135 TI - Management of laryngotracheobronchial sequelae and complications of relapsing polychondritis. AB - Relapsing polychondritis is a rare multisystem disorder of unknown etiology characterized by recurrent inflammation and degeneration of cartilage and connective tissue. Laryngotracheobronchial complications are the most severe manifestations of the disease and present the most challenging management decisions. We present four cases of relapsing polychondritis with laryngotracheobronchial manifestations that illustrate the clinical features and review the treatment options. PMID- 9217136 TI - Coinfection of HPV-11 and HPV-16 in a case of laryngeal squamous papillomas with severe dysplasia. AB - Human papillomavirus (HPV) types 6 and 11 have been associated with benign laryngeal papilloma, while HPV-16 is occasionally associated with laryngeal carcinoma. In this study, a case of laryngeal squamous papillomas with severe dysplasia was evaluated for the presence of HPV infection. The biopsy specimens were taken from a 58-year-old female patient at two different time points 3 months apart. Architecturally, the tumor showed papillary configuration reminiscent of squamous papilloma. Cytologically, the lesion showed morphologic features characteristic of severe squamous epithelial dysplasia. HPV infection was determined by DNA in situ hybridization using type-specific HPV-DNA probes. HPV-11 probes demonstrated homogeneous nuclear staining, suggesting productive viral replication. In contrast, HPV-16 probe produced a speckled pattern, suggesting HPV-16 DNA integration. Normal laryngeal epithelium did not yield specific hybridization. The presence of HPV-11 and HPV-16 was confirmed by PCR using HPV type-specific primers. Immunocytochemical staining was performed to detect Ki-67, a proliferation marker, and p53. Ki-67 expression was demonstrated throughout the whole thickness of epithelium. Staining for p53 was negative. This study suggests that multiple HPV infections can occur in the same lesion and that HPV-16 infection and its DNA integration may contribute to the occurrence of severe dysplasia in the lesion described. PMID- 9217137 TI - Treatment of ventricular dysphonia with botulinum toxin. AB - Ventricular dysphonia, traditionally known as dysphonia plica ventricularis, is a voicing disorder in which the false vocal folds are used as a vibratory source in addition to or instead of the true vocal folds. Traditional treatment of ventricular dysphonia has been voice therapy, which may be slow to produce results if the false fold activity masks an underlying problem of the true folds, is long standing, or has produced hypertrophy of the supraglottic structures. We present seven cases of ventricular dysphonia treated with botulinum toxin injection into the false vocal folds followed by speech therapy. The addition of botulinum toxin to the treatment regimen speeds recovery of normal voicing and allows immediate evaluation of dynamic true vocal fold function by the treating professional. PMID- 9217138 TI - A convenient and efficient moldable dressing for skin grafts. AB - In this paper we present our positive experience with use of Aquaplast thermoplastic as a tie-down dressing for securing and maintaining skin grafts in good position against their nutrient recipient beds. This unique polymer is safe to use, simple to apply, and maintains uniform pressure across the entire surface of the skin graft, no matter how contoured the shape of the anatomical region. PMID- 9217139 TI - Biomechanical effects of e-PTFE implant structure on soft tissue implantation stability: a study in the porcine model. AB - Successful implantation of biocompatible materials depends on physical aspects of its structure. Meshed implants are stable but cannot be easily removed. Nonporous materials are easily removed, but subject to extrusion. We hypothesized that the microporous structure of expanded polytetrafluoroethylene (e-PTFE) would permit limited fibrous ingrowth into the substance of the material, and that tubular implant shape would increase tissue integration while preserving ease of removal. A two-tailed in vivo study was done comparing implant retention, strength of fixation, and removability between tubular and solid-strip e-PTFE implants. Differences in implant retention within tissues were assessed by implanting 396 implants subcutaneously in five swine for observation periods ranging from 3 weeks to 12 months. Strength of implant attachment to host soft tissues was measured at 52 sites by extraction with a tensiometer with forces both parallel and perpendicular to the implant used. Implant porosity was assessed with scanning electron micrography of tubular and solid-strip e-PTFE implants. Measurements of the force and stress tolerances of the implant-tissue interface demonstrated significantly stronger attachment in tubular than strip-shaped implants (P < 0.005). The 11 N (2.75 lb) force sustained by the tubular implant exceeded the 3.4 N (<1 lb) force for the e-PTFE strip by a statistically significant margin on two-tailed Student's t-test (P < 0.005). Even greater forces were tolerated when applied at right angles to the axis of the tubular implant, emulating tissue suspension (21 N, 5.25 lb). The forces and stresses tolerated by both e-PTFE implants far exceeded the fracture stress measured for the implants. Implant extrusion rates were significantly smaller in tubular (0.85%) than in strip-shaped (4.4%) e-PTFE implants (P < 0.05). Standard error of the mean (SEM) demonstrated lesser porosity in tubular than strip implants, suggesting lesser direct tissue attachment. Tubular e-PTFE implant structure facilitates ingrowth of soft tissue through the tube's lumen. This increases the attachment to surrounding soft tissues, increasing fixation strength, decreasing extrusion rate, but still allowing easy removal. These properties may improve clinical applications in facial implantation. PMID- 9217140 TI - Nasal and sinus polyposis in children. AB - Nasal and sinus polyposis in the pediatric population is uncommon and its etiology is unclear. In this 11-year retrospective study, the authors describe the etiologic features and evaluate the effectiveness of endoscopic sinus surgery in 46 children. Patients were divided into three groups according to whether nasal and sinus polyposis was isolated (n = 14), or associated with either asthma (n = 5) or cystic fibrosis (n = 27). An allergy was present in 10% of patients with isolated polyposis, 80% of patients with polyposis associated with asthma, and 22% of patients with polyposis associated with cystic fibrosis. The indications for surgery were disabling symptoms, especially chronic nasal obstruction, rhinorrhea, and mouth breathing, and failure to respond to medical treatment. No surgical complications were encountered. Most patients reported improvement in quality of life with reduction of nasal obstruction in 83% of cases and rhinorrhea in 61%. Minor asymptomatic recurrence (i.e., a few micropolyps localized on the roof of the ethmoid cavity) was observed in 24% of the cases in this series, and major recurrence with the same functional symptoms as before surgery in 12%. However, recurrences were higher in patients with cystic fibrosis, because minor recurrence with no clinical manifestation was observed in 32% of these cases and major recurrence in 16%. Endoscopic sinus surgery must be decided in collaboration with the pediatric and pulmonary physicians, and must be performed skillfully. With a mean follow-up of 3.7 years, results in this series are encouraging. PMID- 9217141 TI - Surgical exposure of the petrous internal carotid artery: practical application for skull base surgery. AB - When exposing the horizontal petrous carotid artery in preparation for intrapetrous carotid bypass, the surgeon has no definite landmarks to localize the perimeter of the cochlea. The results of this study provide a practical, consistent, and safe method to maximize carotid artery exposure while minimizing cochlear injury. We measured the carotid-cochlea distance (mean, 4.3 mm) and the carotid-cochlear angle (mean, 10.8 degrees) in 33 temporal bones in which the extended middle fossa approach had been performed. We correlated this distance to the width of a Sheehy weapon knife, which can be easily measured intraoperatively. Twenty-five temporal bones were imaged prior to surgical exposure using a new computed tomography (CT) protocol that can be used for preoperative assessment of the carotid-cochlear anatomy. The carotid-cochlea distance and carotid-cochlear angle measured on CT are compared with postsurgical measurements. PMID- 9217142 TI - Anterior petrosectomy approach to infraclinoidal basilar artery aneurysms: the emerging role of the neuro-otologist in multidisciplinary management of basilar artery aneurysms. AB - Aneurysms of the basilar artery are uncommon. Historically, because of the central location of these basilar lesions, surgical access has been difficult. Moreover, while this disease and its surgical management inherently carry a high risk of patient morbidity, the presence of neighboring vital neural and vascular structures introduces additional intraoperative challenges. Since 1986 we have employed a transpetrous approach for access to selective aneurysms involving the basilar artery. Removal of the petrous apex has provided an expanded deep window through which infraclinoidal basilar artery aneurysms can be controlled. Reported herein are our results utilizing an anterior petrosectomy approach to the management of infraclinoidal artery aneurysms. PMID- 9217143 TI - A new technique for hypoglossal-facial nerve repair. AB - Hypoglossal reinnervation of the facial nerve may be required after a proximal facial nerve injury. The classic hypoglossal-facial graft procedure involves transection of the donor hypoglossal nerve, resulting in hemiglottic paralysis that, in association with paralysis of other cranial nerves, may cause speech and swallowing difficulties. Multiple lower cranial nerve palsies in conjunction with facial paralysis, as may occur after procedures such as skull base surgery, contraindicate the use of such techniques. The successful use of XII-VII "interposition jump grafts" without hemiglossal weakness has been described However, a prolonged recovery period and weaker facial reanimation have been seen. In order to attain maximum facial reinnervation while preserving hypoglossal function, we have developed a new technique of XII-VII repair. This method involves mobilization of the intratemporal portion of the facial nerve remnant, achieving a single anastomosis with the hypoglossal nerve, which has been partially incised. This technique has been used in three patients to date, with 6 to 11 months follow-up. In all cases facial tone and symmetry have been restored and voluntary facial expression accomplished. The authors conclude that by employing the techniques described highly satisfactory cosmetic and functional results may be expected, without compromising hypoglossal nerve function. PMID- 9217144 TI - Simultaneous treatment with BDNF and CNTF after peripheral nerve transection and repair enhances rate of functional recovery compared with BDNF treatment alone. AB - The objective was to investigate the effects of brain-derived neurotropic factor (BDNF) and ciliary neurotropic factor (CNTF) on peripheral nerve regeneration. Thirty Sprague-Dawley rats underwent left sciatic nerve transection and repair according to three experimental groups: epineurial coaptation (EC), EC with BDNF delivered by an osmotic pump (EC-BDNF), and EC with BDNF and CNTF delivered similarly (EC-BDNF/CNTF). Nerve regeneration was assessed using sciatic functional indices, quantitative histomorphology, and molecular analysis for proteins associated with nerve regeneration. Analysis of variance (ANOVA) comparing all groups at each time point demonstrated significant differences between groups on days 20, 30, 40, 50, 60, and 80. A paired, two-tailed Student's t-test with the Bonferroni correction for multiple comparisons demonstrated that at 40 days postoperatively, animals in the EC-BDNF/CNTF group (n = 7) manifested superior functional recovery compared with those in the EC group (n = 9) and those in the EC-BDNF group (n = 9) (P < 0.001 and P < 0.05, respectively). At 80 days, the animals in both the EC-BDNF (P < 0.01) and EC-BDNF/CNTF (P < 0.05) groups demonstrated greater functional recovery compared with those in the EC group, with no significant difference between the two factor groups at the endpoint. Morphometric analysis demonstrated that nerves from animals in the EC BDNF/CNTF group had the largest mean axon diameters as compared with those from the EC (proximal: P < 0.001, distal: P < 0.05) and EC-BDNF (proximal: P < 0.01) groups. No significant differences were seen in nerve cross-sectional area. In distal nerve segments, Western blot analysis revealed that expression of myelin associated glycoprotein was higher than control for the EC group and lower than control for both the EC-BDNF and EC-BDNF/CNTF groups. We conclude that BDNF/CNTF combined treatment increases the early rate of functional sciatic nerve regeneration over treatment with BDNF alone, although the degree of maximal recovery was similar at the conclusion of the experiment. PMID- 9217145 TI - CNRS returns fire on ethics review panel. PMID- 9217146 TI - AIDS 'cure' researchers guilty of misconduct. PMID- 9217147 TI - Just follow the acid chain. PMID- 9217148 TI - Apoptosis. Placing death under control. PMID- 9217149 TI - Alfred Hershey (1908-97) PMID- 9217150 TI - Lifetime of plasma cells in the bone marrow. PMID- 9217151 TI - Importance of ancestral DNA ages. PMID- 9217152 TI - Proximal-distal axis formation in the Drosophila leg. AB - Limb development requires the formation of a proximal-distal axis perpendicular to the main anterior-posterior and dorsal-ventral body axes. The secreted signalling proteins Decapentaplegic and Wingless act in a concentration-dependent manner to organize the proximal-distal axis. Discrete domains of proximal-distal gene expression are defined by different thresholds of Decapentaplegic and Wingless activities. Subsequent modulation of the relative sizes of these domains by growth of the leg is required to form the mature pattern. PMID- 9217153 TI - Properties of ideal composite knots. AB - The shortest tube of constant diameter that can form a given knot represents the 'ideal' form of the knot. Ideal knots provide an irreducible representation of the knot, and they have some intriguing mathematical and physical features, including a direct correspondence with the time-averaged shapes of knotted DNA molecules in solution. Here we describe the properties of ideal forms of composite knots-knots obtained by the sequential tying of two or more independent knots (called factor knots) on the same string. We find that the writhe (related to the handedness of crossing points) of composite knots is the sum of that of the ideal forms of the factor knots. By comparing ideal composite knots with simulated configurations of knotted, thermally fluctuating DNA, we conclude that the additivity of writhe applies also to randomly distorted configurations of composite knots and their corresponding factor knots. We show that composite knots with several factor knots may possess distinct structural isomers that can be interconverted only by loosening the knot. PMID- 9217154 TI - The dynamics of partially extended single molecules of DNA. AB - The behaviour of an isolated polymer floating in a solvent forms the basis of our understanding of polymer dynamics. Classical theories describe the motion of a polymer with linear equations of motion, which yield a set of 'normal modes', analogous to the fundamental frequency and the harmonics of a vibrating violin string. But hydrodynamic interactions make polymer dynamics inherently nonlinear, and the linearizing approximations required for the normal-mode picture have therefore been questioned. Here we test the normal-mode theory by measuring the fluctuations of single molecules of DNA held in a partially extended state with optical tweezers. We find that the motion of the DNA can be described by linearly independent normal modes, and we have experimentally determined the eigenstates of the system. Furthermore, we show that the spectrum of relaxation times obeys a power law. PMID- 9217155 TI - Evolution of genetic redundancy. AB - Genetic redundancy means that two or more genes are performing the same function and that inactivation of one of these genes has little or no effect on the biological phenotype. Redundancy seems to be widespread in genomes of higher organisms. Examples of apparently redundant genes come from numerous studies of developmental biology, immunology, neurobiology and the cell cycle. Yet there is a problem: genes encoding functional proteins must be under selection pressure. If a gene was truly redundant then it would not be protected against the accumulation of deleterious mutations. A widespread view is therefore that such redundancy cannot be evolutionarily stable. Here we develop a simple genetic model to analyse selection pressures acting on redundant genes. We present four cases that can explain why genetic redundancy is common. In three cases, redundancy is even evolutionarily stable. Our theory provides a framework for exploring the evolution of genetic organization. PMID- 9217156 TI - Distinct cortical areas associated with native and second languages. AB - The ability to acquire and use several languages selectively is a unique and essential human capacity. Here we investigate the fundamental question of how multiple languages are represented in a human brain. We applied functional magnetic resonance imaging (fMRI) to determine the spatial relationship between native and second languages in the human cortex, and show that within the frontal lobe language-sensitive regions (Broca's area), second languages acquired in adulthood ('late' bilingual subjects) are spatially separated from native languages. However, when acquired during the early language acquisition stage of development ('early' bilingual subjects), native and second languages tend to be represented in common frontal cortical areas. In both late and early bilingual subjects, the temporal-lobe language-sensitive regions (Wernicke's area) also show effectively little or no separation of activity based on the age of language acquisition. This discovery of language-specific regions in Broca's area advances our understanding of the cortical representation that underlies multiple language functions. PMID- 9217157 TI - Representation of motion boundaries in retinotopic human visual cortical areas. AB - Edges are important in the interpretation of the retinal image. Although luminance edges have been studied extensively, much less is known about how or where the primate visual system detects boundaries defined by differences in surface properties such as texture, motion or binocular disparity. Here we use functional magnetic resonance imaging (fMRI) to localize human visual cortical activity related to the processing of one such higher-order edge type: motion boundaries. We describe a robust fMRI signal that is selective for motion segmentation. This boundary-specific signal is present, and retinotopically organized, within early visual areas, beginning in the primary visual cortex (area V1). Surprisingly, it is largely absent from the motion-selective area MT/V5 and far extrastriate visual areas. Changes in the surface velocity defining the motion boundaries affect the strength of the fMRI signal. In parallel psychophysical experiments, the perceptual salience of the boundaries shows a similar dependence on surface velocity. These results demonstrate that information for segmenting scenes by relative motion is represented as early as V1. PMID- 9217158 TI - The synaptic activation of kainate receptors. AB - L-Glutamate, the principal excitatory neurotransmitter in the vertebrate central nervous system, acts on three classes of ionotropic glutamate receptors, named after the agonists AMPA, NMDA and kainate. AMPA receptors are known to mediate fast synaptic responses and NMDA receptors to mediate slow synaptic responses at most excitatory synapses in the brain. Kainate receptors are formed from a separate set of genes (GluR5-7, KA-1 and KA-2) and are widely distributed throughout the brain. They are implicated in epileptogenesis and cell death. However, the physiological functions of kainate receptors are not known. The development of 2,3-benzodiazepine antagonists that are selective for AMPA receptors enables kainate receptors to be specifically activated by exogenous ligands, such as kainate. Here we demonstrate that high-frequency stimulation of mossy fibres in rat hippocampal slices, in the presence of the highly selective AMPA receptor antagonist GYKI 53655 plus NMDA- and GABA-receptor antagonists, activates an inward current in CA3 neurons that has a pharmacology typical of kainate receptors. The finding that kainate receptors can be activated synaptically adds to the diversity of information transfer at glutamatergic synapses. PMID- 9217159 TI - Kainate receptors mediate a slow postsynaptic current in hippocampal CA3 neurons. AB - Glutamate, the neurotransmitter at most excitatory synapses in the brain, activates a variety of receptor subtypes that can broadly be divided into ionotropic (ligand-gated ion channels) and metabotropic (G-protein-coupled) receptors. Ionotropic receptors mediate fast excitatory synaptic transmission, and based on pharmacological and molecular biological studies are divided into NMDA and non-NMDA subtypes. The non-NMDA receptor group is further divided into AMPA and kainate subtypes. Virtually all fast excitatory postsynaptic currents studied so far in the central nervous system are mediated by the AMPA and NMDA subtypes of receptors. Surprisingly, despite extensive analysis of their structure, biophysical properties and anatomical distribution, a synaptic role for kainate receptors in the brain has not been found. Here we report that repetitive activation of the hippocampal mossy fibre pathway, which is associated with high-affinity kainate binding and many of the kainate receptor subtypes, generates a slow excitatory synaptic current with all of the properties expected of a kainate receptor. This activity-dependent synaptic current greatly augments the excitatory drive of CA3 pyramidal cells. PMID- 9217160 TI - Muscle force is generated by myosin heads stereospecifically attached to actin. AB - Muscle force is generated by myosin crossbridges interacting with actin. As estimated from stiffness and equatorial X-ray diffraction of muscle and muscle fibres, most myosin crossbridges are attached to actin during isometric contraction, but a much smaller fraction is bound stereospecifically. To determine the fraction of crossbridges contributing to tension and the structural changes that attached crossbridges undergo when generating force, we monitored the X-ray diffraction pattern during temperature-induced tension rise in fully activated permeabilized frog muscle fibres. Temperature jumps from 5-6 degrees C to 16-19 degrees C initiated a 1.7-fold increase in tension without significantly changing fibre stiffness or the intensities of the (1,1) equatorial and (14.5 nm)(-1) meridional X-ray reflections. However, tension rise was accompanied by a 20% decrease in the intensity of the (1,0) equatorial reflection and an increase in the intensity of the first actin layer line by approximately 13% of that in rigor. Our results show that muscle force is associated with a transition of the crossbridges from a state in which they are nonspecifically attached to actin to one in which stereospecifically bound myosin crossbridges label the actin helix. PMID- 9217161 TI - Inhibition of death receptor signals by cellular FLIP. AB - The widely expressed protein Fas is a member of the tumour necrosis factor receptor family which can trigger apoptosis. However, Fas surface expression does not necessarily render cells susceptible to Fas ligand-induced death signals, indicating that inhibitors of the apoptosis-signalling pathway must exist. Here we report the characterization of an inhibitor of apoptosis, designated FLIP (for FLICE-inhibitory protein), which is predominantly expressed in muscle and lymphoid tissues. The short form, FLIPs, contains two death effector domains and is structurally related to the viral FLIP inhibitors of apoptosis, whereas the long form, FLIP(L), contains in addition a caspase-like domain in which the active-centre cysteine residue is substituted by a tyrosine residue. FLIPs and FLIP(L) interact with the adaptor protein FADD and the protease FLICE, and potently inhibit apoptosis induced by all known human death receptors. FLIP(L) is expressed during the early stage of T-cell activation, but disappears when T cells become susceptible to Fas ligand-mediated apoptosis. High levels of FLIP(L) protein are also detectable in melanoma cell lines and malignant melanoma tumours. Thus FLIP may be implicated in tissue homeostasis as an important regulator of apoptosis. PMID- 9217162 TI - Tom5 functionally links mitochondrial preprotein receptors to the general import pore. AB - Most mitochondrial proteins are synthesized as preproteins on cytosolic polysomes and are subsequently imported into the organelle. The mitochondrial outer membrane contains a multisubunit preprotein translocase (Tom) which has receptors on the cytosolic side and a general import pore (GIP) in the membrane. Tom20 Tom22 and Tom70-Tom37 function as import receptors with a preference for preproteins that have amino-terminal presequences or internal targeting information, respectively. Tom40 is an essential constituent of the GIP, whereas Tom6 and Tom7 modulate the assembly and dissociation of the Tom machinery. Here we report the identification of Tom5, a small subunit that has a crucial role importing preproteins destined for all four mitochondrial subcompartments. Tom5 has a single membrane anchor and a cytosolic segment with a negative net charge, and accepts preproteins from the receptors and mediates their insertion into the GIP. We conclude that Tom5 represents a functional link between surface receptors and GIP, and is part of an 'acid chain' that guides the stepwise transport of positively charged mitochondrial targeting sequences. PMID- 9217163 TI - X-chromosome-counting mechanisms that determine nematode sex. AB - Sex is determined in Caenorhabditis elegans by an X-chromosome-counting mechanism that reliably distinguishes the twofold difference in X-chromosome dose between males (1X) and hermaphrodites (2X). This small quantitative difference is translated into the 'on/off' response of the target gene, xol-1, a switch that specifies the male fate when active and the hermaphrodite fate when inactive. Specific regions of X contain counted signal elements whose combined dose sets the activity of xol-1. Here we ascribe the dose effects of one region to a discrete, protein-encoding gene, fox-1. We demonstrate that the dose-sensitive signal elements on chromosome X control xol-1 through two different molecular mechanisms. One involves the transcriptional repression of xol-1 in XX animals. The other uses the putative RNA-binding protein encoded by fox-1 to reduce the level of xol-1 protein. These two mechanisms of repression act together to ensure the fidelity of the X-chromosome counting process. PMID- 9217165 TI - Oligosaccharide sequencing technology. PMID- 9217166 TI - Diagnosis of cobalamin deficiency: the old and the new. PMID- 9217167 TI - Human eosinophils: their accumulation, activation and fate. PMID- 9217168 TI - Long-term stromal cultures produce dendritic-like cells. AB - A stromal cell-dependent long-term culture (LTC) system has been developed from spleen which continuously generates non-adherent cells with dendritic cell (DC) characteristics. Bioassays using factor-dependent cell lines have revealed that both spleen and thymic stromal cultures secrete interleukin (IL)-3 and IL-6-like growth factors. Conditioned medium from LTC also contains factors which appear to be unrelated to IL-3 and induces growth of stromal cells from bone marrow. Non adherent cells generated in LTC were not T or B lymphoid cells or granulocytes, nor were mast cells detectable. Morphological and electron microscopic examination has also excluded the presence of macrophages (Mo). Cells with DC morphology have been detected by both light and electron microscopy. The majority of cells in the non-adherent population were found to have multiple membrane pseudopodia, with a small acentric nucleus. These appear to be the precursors of DC. They expressed cell surface markers detectable with DC-specific antibodies and antibody specific for major histocompatibility complex Class II molecules. A proportion of cells also expressed myeloid markers, but since this expression was not supported by histochemical staining for myeloperoxidase or non-specific esterase, it was concluded that the cells produced are not typical of the myeloid lineage. Cells generated in LTC were shown to be potent stimulators for allogeneic and syngeneic MLR and for antigen-specific T-cell proliferation. PMID- 9217169 TI - Differential modulation of IL-1-induced endothelial adhesion molecules and transendothelial migration of granulocytes by G-CSF. AB - Granulocyte colony stimulating factor (G-CSF) is widely used for mobilization of haemopoietic stem cells into the peripheral blood. However, little is known about the mechanisms involved in mobilization and the immune modulatory effects of this growth factor. In this report we show that G-CSF down-regulated intercellular adhesion molecule 1 (ICAM-1) induced by Interleukin-1 (IL-1) on human endothelial cells. Interestingly, the G-CSF-mediated down-modulation of IL-1-induced ICAM-1 appeared to be biphasic. In pharmacological concentrations (> 300 ng/ml), and in dose ranges of plasma G-CSF levels above that of nonfebrile healthy individuals (30 pg/ml), a significant decrease in surface ICAM-1 could be observed. This could be explained, at least in part, by an increased autocrine G-CSF production by endothelial cells in response to IL-1 and exogenous G-CSF. In contrast to ICAM 1, IL-1-triggered VCAM-1 expression was superinduced by G-CSF with the optimal concentration of 30 pg/ml. To evaluate the functional significance of these findings, 51Cr adhesion assays with peripheral blood mononuclear cells (PBMC) or granulocytes known to lack the VCAM-1 counter-receptor very late antigen 4 (VLA 4) and IL-1-stimulated endothelial cells, in the presence or absence of G-CSF, were performed. G-CSF could not inhibit the IL-1-induced adhesion of PBMC to endothelial cells, which may be due to the differential adhesion molecule modulation. In contrast, granulocyte adhesion induced by IL-1 could effectively be blocked by co-incubation with G-CSF. Finally, G-CSF also inhibited transendothelial migration of granulocytes through IL-1-activated endothelial cells in a concentration-dependent manner. PMID- 9217170 TI - Bone mineralization and turnover in children with congenital neutropenia, and its relationship to treatment with recombinant human granulocyte-colony stimulating factor. AB - Bone mineral content (BMC) of the radius was measured using single photon absorptiometry (SPA) in nine children with congenital neutropenia. Five had normal values. Two children with severe congenital neutropenia (SCN) had low BMC, and two boys with Schwachman syndrome had biochemistry suggestive of rickets. PMID- 9217171 TI - The role of immunoglobulin G and fibrinogen in platelet aggregation by Streptococcus sanguis. AB - Previous work has shown that the type strain of Streptococcus sanguis, NCTC 7863, induces aggregation of normal platelets by a complement-dependent mechanism. We investigated the roles of IgG and fibrinogen in the aggregation process. Plasma depleted of IgG by passage through protein A-sepharose failed to support platelet aggregation, as did plasma absorbed at 0 degrees C with whole bacteria. However, absorption of plasma with a non-aggregating strain of S. sanguis, SK96, did not remove aggregating activity for NCTC 7863. Supplementing 0 degrees C-absorbed plasma with purified IgG restored the aggregation supporting activity. A monoclonal antibody to the Fc gammaRII receptor inhibited platelet aggregation by the bacteria, indicating a requirement for bacteria-IgG complexes interacting with the Fc receptor in platelet aggregation. There was a lag time to the onset of platelet aggregation of 7-19 min depending upon the platelet donor, but the length of this lag did not correlate with either total IgG concentration recognizing NCTC 7863 in subjects' plasma, or the concentration any of the four IgG subclasses or with IgG avidity levels. Fibrinogen was shown to bind rapidly to the bacterial cell surface. Monoclonal antibody to GPIIb/IIIa, RGDS peptide, and a specific antagonist for the platelet fibrinogen receptor, GPIIb/IIIa, FK633, inhibited platelet aggregation by NCTC 7863, indicating that platelet aggregation is fibrinogen dependent. These data suggest that platelet aggregation by some strains of S. sanguis requires multiple stimuli/agonists, including IgG Fc receptor interaction, complement and fibrinogen. PMID- 9217172 TI - Flow cytometric analysis of platelets from children with the Wiskott-Aldrich syndrome reveals defects in platelet development, activation and structure. AB - The pathophysiology of platelet dysfunction in the Wiskott-Aldrich immune deficiency syndrome (WAS) remains unclear. Using flow cytometry, we have characterized the functional properties of platelets from 10 children with WAS. Patients with WAS had thrombocytopenia, small platelets, increased platelet associated IgG and reduced platelet-dense granule content. Levels of reticulated 'young' platelets were normal in the WAS patients. Although the mean numbers of platelet glycoprotein (GP) Ib, GPIIbIIIa and GPIV molecules per platelet appeared lower in WAS patients than in healthy controls, analysis of similar-sized platelets revealed the mean number of GPIb molecules per platelet to be comparable in patients and normal controls. Surface GPIIbIIIa and GPIV expression was, however, significantly lower on the WAS platelets than on normal platelets. Compared with normal platelets, WAS platelets showed a reduced ability to modulate GPIIbIIIa expression following thrombin stimulation. In addition, thrombin- and ADP-induced expression of CD62P and CD63 was defective in WAS platelets. Phallacidin staining of the WAS platelets revealed less F-actin content than in normal platelets. Together, these data suggest that the reduced platelet number and function in WAS reflects, at least in part, a defect in bone marrow production as well as an intrinsic platelet abnormality. PMID- 9217173 TI - Detection of drug-dependent, platelet-reactive antibodies by solid-phase red cell adherence assays. AB - We developed a simple modification of the solid-phase red cell adherence (SPRCA) assay system that can be used to identify drug-dependent platelet antibodies (DDPAs) reactive by either the hapten or immune complex reaction mechanisms. Between January 1994 and August 1996 we tested sera from 173 patients [123 (71%) with unexplained thrombocytopenia and 50 (29%) because of poor responses to platelet transfusions not explicable by alloimmunization or the clinical situation] for DDPAs possibly associated with the receipt of 61 different drugs. We correlated positive results with patients' clinical courses. DDPAs were identified in samples from 138 (80%) of the patients tested. Antibodies reactive only by the hapten mechanism were identified in 51 (37%) of those sera exhibiting positive reactions. The clinical courses of 108 (78%) patients were evaluable. Discontinuation of the implicated drug(s) resulted in prompt (<5 d) resolution of the thrombocytopenia or improvement in response to transfusion in all of these patients. In four cases thrombocytopenia returned upon re-exposure to the implicated drug. This adaptation of SPRCA provides a simple means of investigating the possibility of DDPAs and documents a higher frequency of these antibodies than has previously been suspected. PMID- 9217174 TI - Proteolytic degradation of high molecular weight kininogen in acute thrombotic thrombocytopenic purpura. AB - Thrombotic thrombocytopenic purpura (TTP) is a disorder characterized by thrombocytopenia and schistocytic haemolytic anaemia. The majority of patients have normal coagulation parameters including the activated partial thromboplastin time (APTT). An intracellular cysteine protease, calpain, has been found in the plasma of many patients with acute TTP, and we hypothesize that it participates in the pathogenesis of the disorder. However, certain aspects of the disorder remain unresolved. For example, high molecular weight kininogen (HK) is one of the primary plasma inhibitors of calpain, and it also can act as a substrate for calpain. Consequently, one might anticipate that the HK could be defective or altered in TTP. In this report we describe the analysis of HK in plasma from live patients with acute TTP and following recovery. The HK was studied immunogenically and functionally. We observed that the HK in plasma samples from patients with acute TTP was proteolysed. This degradation was not observed in remission samples from the same patients. However, both acute and remission TTP samples had normal HK coagulant activity (acute samples, x = 0.84 +/- 0.26 U/ml; remission samples, x = 0.89 +/- 0.21 U/ml; control samples, x = 0.87 +/- 0.05 U/ml). Although the TTP plasmas were able to inhibit calpain activity, less inhibition activity was found in the acute samples (acute: mean inhibition 71 +/- 2.4%; remission: mean 92 +/- 2.1%; control samples: mean 93 +/- 5.4%; P < 0.001). Normal HK treated with calpain also had reduced calpain inhibition capacity (mean 58%). This study suggests that although HK is proteolysed during acute TTP, the proteolysis occurs without a major loss in the coagulation function or depletion of the protease inhibitory activity of HK. PMID- 9217175 TI - Antiphospholipid antibody: functional specificity for inhibition of prothrombin activation by the prothrombinase complex. AB - The antiphospholipid syndrome (APS) is associated with production of autoantibodies with lupus anticoagulant (LA) activity. These antibodies cause prolongation of in vitro clotting tests by inhibition of the conversion of prothrombin to thrombin in the presence of anionic phospholipid (PL). The extent to which this inhibition reflects antibody binding to, or functional inhibition of, phospholipids alone, prothrombin alone, or a prothrombin-phospholipid complex is pertinent to our understanding of the pathophysiology of this syndrome. Immunoglobulin fractions (IgG) from 18 patients with LA activity were tested for inhibitory activity against prothrombin activation by either factor Xa, in a purified prothrombinase system, or by purified fractions of snake venoms (E. carinatus, E. multisquamatus) which cleave prothrombin at the same initial site as the prothrombinase complex but do not require anionic phospholipid as a cofactor. Parallel testing of the same IgG samples for prothrombin binding by immunoassay was performed. Although all IgG samples inhibited the prothrombinase reaction, only three exhibited any inhibition of venom protease prothrombin activation in either the presence or absence of PL. Only one sample exhibited prothrombin binding by Western blot. These results suggest that lupus anticoagulant antibodies are heterogenous and that many, if not most, of the autoantibody populations responsible for LA activity impair prothrombin activation by interaction either with phospholipid alone or with a restricted range of prothrombin-phospholipid epitopes expressed by prothrombin only as part of the intact prothrombin-prothrombinase complex. PMID- 9217176 TI - Epidemiology of coagulation factors, inhibitors and activation markers: the Third Glasgow MONICA Survey. I. Illustrative reference ranges by age, sex and hormone use. AB - Coagulation factor activity (fibrinogen, VII, VIII and IX), coagulation inhibitor activity (antithrombin, protein C, protein S), and coagulation activation markers (prothrombin fragment F1, 2; thrombin-antithrombin complexes) were measured in 747 men and 817 women aged 25-74 years, randomly sampled from the north Glasgow population in the Third MONICA Survey. Significant effects of age, sex, menopause and hormone use were observed and specific reference ranges are presented to illustrate these effects. Significant correlations were observed between several coagulation factors and inhibitors. Increased levels of factors VII, VIII and IX and decreased levels of protein C were associated with increased coagulation activation. In general, increases in coagulation factors with age were greater than increases in coagulation inhibitors, especially in men; this imbalance may favour increased coagulation activation and hence increased thrombotic risk with age. PMID- 9217178 TI - A comparison between two activated protein C resistance methods as routine diagnostic tests for factor V Leiden mutation. AB - The most common commercially available test measuring activated protein C (APC) resistance relies on the the anticoagulant response to added APC in an activated partial thromboplastin time (APTT) based method. Another method is a Russell Viper venom time (RVVT) based system. To improve the specificity for factor V Leiden of the APTT based method, pre-dilution of test plasma in FV-deficient plasma has recently been recommended. In this study we tested the relative suitabilities of the APTT-based system, the RVVT-based system and their corresponding assays modified by pre-dilution in FV-deficient plasma, for screening asymptomatic subjects, a group of thrombophilic patients (in particular those with low APC ratios), patients on oral anticoagulants, and patients with lupus anticoagulant (LAC). We found the RVVT-based assay to be superior to the APTT-based method in the separation of normals from those with FV Leiden mutation both in asymptomatic subjects and in the thrombophilic patient group. Both modified assays demonstrated a sensitivity and specificity of 100% for FV Leiden, as verified by genotyping in asymptomatic subjects, thrombophilic patients and patients on oral anticoagulants, with the modified RVVT-based assay giving better separation between normals and FV Leiden. Inhibition of phospholipid-dependent coagulation by LAC antibodies rendered the APTT-based system less suitable than the phospholipid-rich RVVT-based one, and as nine of the 20 LAC-positive patients were on warfarin, we showed only the modified RVVT assay to be a reliable predictor of factor V Leiden in this patient group. PMID- 9217177 TI - Epidemiology of coagulation factors, inhibitors and activation markers: The Third Glasgow MONICA Survey. II. Relationships to cardiovascular risk factors and prevalent cardiovascular disease. AB - Coagulation factor activity (fibrinogen, VII, VIII and IX), coagulation inhibitor activity (antithrombin, protein C, protein S), and coagulation activation markers (prothrombin fragment F1, 2; thrombin-antithrombin complexes) were measured in 746 men and 816 women aged 25-74 years, randomly sampled from the north Glasgow population in the Third MONICA Survey. After age-adjustment, significant associations with cardiovascular risk factors were observed. Serum cholesterol and triglyceride were associated with increases in factors VII and IX, as well as antithrombin, protein C and protein S; and with increased fibrinogen and factor VIII in women. Apart from factor VIII (related to blood pressure in men, but not in women), similar associations were observed for blood pressure and body mass index. Smoking status and/or smoking markers were related to fibrinogen, factor IX, antithrombin and protein S. Alcohol intake was related to protein S, and inversely to fibrinogen and antithrombin in men. Low social class was associated with fibrinogen, factor VIII, factor IX, and with antithrombin, protein S, and low protein C in men. Serum vitamin C was associated inversely with coagulation factors and coagulation inhibitors. The only associations of activation markers were with low serum vitamin C, and with alcohol consumption and low social class in men. Prevalent cardiovascular disease was associated only with fibrinogen. These associations of coagulation factors and inhibitors with cardiovascular risk factors are plausibly relevant to thrombotic risk in cardiovascular disease. In general, 'worse' values of risk factors are associated with increased plasma levels of both coagulation factors and inhibitors, without significant increase in coagulation activation markers. However, the association of lower serum vitamin C with increased coagulation activation markers is of potential therapeutic interest. PMID- 9217179 TI - The VITA project: C677T mutation in the methylene-tetrahydrofolate reductase gene and risk of venous thromboembolism. AB - We evaluated the hypothesis that a common polymorphism of the methylenetetrahydrofolate reductase gene (C677T), which results in increased levels of plasma homocysteine, may be a putative risk factor for venous thromboembolism (VT). Sixty-five cases of VT and 130 controls, both identified within the framework of an epidemiologic survey on thrombophilia, the Vicenza Thrombophilia and Arteriosclerosis (VITA) Project, were genotyped for the mutation. No increased risk of VT was found in carriers of the mutation. We conclude that screening for the C677T mutation of the methylenetetrahydrofolate reductase gene should not be recommended in unselected patients with VT. PMID- 9217180 TI - Prevalence of anti-FVIII antibodies in severe haemophilia A patients with inversion of intron 22. AB - The aim of our study was to investigate whether haemophilia A patients with inversion of intron 22 are at high risk for non-inhibitory anti-FVIII antibodies development detected by ELISA. It is known that patients with severe forms of haemophilia A are more likely to develop anti-FVIII antibodies. The incidence of inhibitory anti-FVIII antibodies in patients with factor VIII gene inversion has been extensively evaluated, but if this defect has to be considered a predisposing factor is still debatable. Non-inhibitory anti-FVIII antibodies are attracting interest, due to the potential influence on FVIII half-life. Our data show that FVIII gene inversion was a major predisposing factor for anti-FVIII antibodies development. PMID- 9217181 TI - Prognostic features of asymptomatic multiple myeloma. AB - Approximately 20% of patients with multiple myeloma are recognized by chance without significant symptoms. In order to prevent morbidity with timely therapy, reliable criteria are needed that distinguish those likely to show early or late disease progression. Multiple clinical features were assessed in 101 consecutive, asymptomatic and previously untreated patients. Patients with one or more lytic bone lesions were excluded because this feature had been found previously to be associated with early progression. Multivariate analysis indicated that only serum myeloma globulin > 30 g/l, IgA protein type, and Bence Jones protein excretion > 50 mg/d remained as significant independent variables. The presence of two or more of these features signified high-risk disease with early progression (median 17 months) whereas the absence of any adverse variable was associated with prolonged stability (median 95 months) (P < 0.01). Magnetic resonance (MR) imaging of the spine was useful only in patients with one adverse feature and an intermediate time to progression (median 39 months). An abnormal pattern (40% of patients) helped to distinguish patients with an imminent complication from those with more stable disease. Because a serious complication (fracture, hypercalcaemia) occurred in 35% of patients with early disease progression, chemotherapy seems justified for selected patients with asymptomatic disease at diagnosis. The remaining patients were at such low risk for progression (median 6 years) that they may be followed safely at long intervals without treatment. PMID- 9217182 TI - Distribution of T-cell signalling molecules in human myeloma. AB - It is controversial whether altered levels of TCR/CD3-associated signalling molecules play a role in the T-cell dysfunction of cancer patients. In multiple myeloma (MM), peripheral blood T (PBT) lymphocytes are functionally impaired by prolonged exposure to tumour cells, and so we investigated the organization of the TCR/CD3-associated signal transduction machinery. The aim of this study was two-fold: first, to investigate the levels of CD3zeta, p56(lck), p59(fyn), ZAP 70, protein kinase C-alpha (PKC-alpha) and phospholipase C-gamma (PLC-gamma) in MM PBT cells; second, to determine whether levels of expression were correlated with clinical or prognostic factors. Forty-four MM patients were studied and 25 age-matched normal donors served as controls. On average, PKC-alpha was the only significantly decreased (P<0.001) signalling molecule, whereas levels of CD3zeta, p56(lck), p59(fyn), PLC-gamma and ZAP-70 were not statistically different. However, there was wide variation between individual patients, and levels for each single protein also varied. A 75% or greater decrease in protein expression was observed, ranging from 8% (p59(fyn)) to 68% (PCK-alpha) of MM patients. When patients were grouped according to the cut-off values of prognostic factors such as the serum levels of C reactive protein (CRP), beta2-microglobulin (beta2M), neopterin (NPT) and the labelling index (LI%) of bone marrow (BM) plasma cells, the only difference observed was the lower PKC-alpha expression in patients with high serum NPT values. None of the T-cell signalling molecule levels was affected by the duration of tumour exposure, calculated on the number of years and/or months that had elapsed since diagnosis, or by disease status. In conclusion, there was a significant decrease of PCK-alpha in MM T cells; however, neither this decrease nor the heterogenous levels of the other T-cell signalling molecules were clearly correlated with prognosis, duration of tumour exposure, and disease status. PMID- 9217183 TI - CD56+ NK lymphomas: clinicopathological features and prognosis. AB - The surface molecule CD56 marks a category of malignant lymphoma of putative natural killer (NK) cell origin. We conducted a retrospective analysis of 24 cases of CD56+ NK lymphoma/leukaemia to define the clinicopathologic and prognostic features of this specific group of lymphomas. 56 cases of nasal lymphomas and 204 cases with an initial diagnosis of peripheral T-cell lymphoma were retrospectively analysed. To specifically examine lymphomas of putative NK origin, only those that were negative for surface expression of CD3 but positive for CD56 were analysed. 24 cases were identified. The initial predominant sites of involvement were nasal (n = 18), palate (n = 1), nodal (n = 1) and multi-organ (n = 4). Clinically, in patients with disease localized to one anatomical site (n = 20), most had symptoms confined to the nose, with a high percentage in early stage (I: 91%; IV: 9%). The marrow was not involved in any of these cases. However, patients with multi-organ involvement at presentation (n = 4) behaved differently. All presented acutely with pancytopenia, hepatosplenomegaly, and marrow infiltration with haemophagocytosis. A leukaemic phase was observed in one case. Anthracycline containing combination chemotherapy resulted in complete remission in 75% of patients with localized disease, but only in 25% with multi organ involvement. The median survival of patients with localized disease was 12 months, compared with 2 months in the multi-organ group (P = 0.06); the disease free survival was significantly better in the former (P < 0.01). The overall median survival of all patients was still poor at 11 months. We conclude that CD56+ NK lymphomas could be divided into two main patterns of disease presentations: localized (predominantly nasal), and multi-organ involvement. Each has different clinicopathologic and prognostic features. Conventional chemotherapy appeared ineffective for the majority of patients, and innovative treatment modalities are needed to improve outcome. PMID- 9217184 TI - Immunoglobulin heavy chain diversity genes rearrangement pattern indicates that MALT-type gastric lymphoma B cells have undergone an antigen selection process. AB - Gastric MALT lymphoma usually develops from chronic gastritis, the vast majority of which (>90%) is associated with Helicobacter pylori infection. We sequenced the third complementarity determining region (CDR3) of immunoglobulin heavy chain genes in 19 gastric MALT lymphoma clones to determine the pattern of variable (V), diversity (D) and joining (J) gene utilization during immunoglobulin gene rearrangement. DNA was extracted from paraffin-embedded sections and the rearranged CDR3 regions were amplified using a semi-nested polymerase chain reaction (with primers complementary to the conserved framework-three segment of the variable region [FR3A] and J regions). The DNA used for cloning and sequencing was obtained after purification of monoclonal bands excised from polyacrylamide gels. The N-D-N region specific to each clone was compared with known germline D sequences. Similarly to that observed in normal and leukaemic B cells, our series of gastric MALT lymphomas showed apparent preferential utilization of genes from the DXP family. In two cases no similarity between the CDR3 nucleotide sequences of the neoplastic clones and the known germline D sequences could be found. In 10/19 analysed alleles the lymphoma B-cell clones appeared to contain two D gene segments (D-D recombination), a rare occurrence in normal individuals but one which has been described as a significant event in the determination of idiotype expression and antigen-binding affinity. Remarkably, despite the use of different D and J segments, the resultant amino acid sequences matched in two patients, suggesting the presence of a common selecting antigen. The observed pattern of D gene rearrangement suggests that MALT lymphoma B-cell clones have undergone antigen selection, which seems to indicate that the antigen stimulation plays a pivotal role in the development of the lymphoma. PMID- 9217185 TI - Heterogenous expression of bcr-abl fusion mRNA in a patient with Philadelphia chromosome-positive acute lymphoblastic leukaemia. AB - We performed molecular and cytogenetic analysis on a 56-year-old woman with Philadelphia chromosome (Ph1)-positive acute lymphoblastic leukaemia (ALL) having two types of major and minor bcr-abl (M-bcr-abl, m-bcr-abl/fusion mRNA at diagnosis. In the course of her disease, unexpected heterogenous bcr-abl fusion mRNA was detected by sequential analysis using the reverse transcription and polymerase chain reaction (RT-PCR). The RT-PCR analysis showed both M-bcr-abl (b2 a2 type) and m-bcr-abl at diagnosis. Although b2-a2 type M-bcr-abl disappeared after complete remission (CR) was achieved with intensive induction chemotherapy, expression of both m-bcr-abl and a new type of M-bcr-abl mRNA (b1-a2 type), which may have been produced through splicing out of exon b2, was detected in the early stage of CR. The leukaemic cells contained these heterogenous bcr-abl mRNAs in both the CR and relapsed state, and showed more stable expression of the m-bcr abl type mRNA than that of the b2-a2 type. These findings of heterogenous bcr-abl mRNA due to alternative or missplicing during the disease course in the present ALL case may provide important evidence of disease progression. PMID- 9217186 TI - Myeloperoxidase gene expression in acute lymphoblastic leukaemia. AB - The FAB classification of acute leukaemias is based on the light microscopic detection of myeloperoxidase (MPO) activity in blast cells. Cases with MPO activity in > or = 3% of blasts are classified as acute myeloid leukaemia. We describe two cases of acute leukaemia in which the blast cells had lymphoid morphology, ultrastructure, immunophenotype and molecular rearrangements, but expressed significant levels of the MPO gene (MPO mRNA) by reverse transcription polymerase chain reaction (RT-PCR), in the absence of translation to protein. Both cases presented a short remission duration. We discuss the incidence, features and outcome of MPO mRNA positive acute lymphoblastic leukaemia (ALL). PMID- 9217187 TI - Lack of clonal BCRA2 gene deletion on chromosome 13 in chronic lymphocytic leukaemia. AB - Chromosome 13q deletion is among the most common cytogenetic abnormalities in chronic lymphocytic leukaemia (CLL). We investigated the 13q14.3 deletion in 44 CLL patients by Southern blotting following purification of clonal B CLL cells to >90%. Two sets of probes were used to investigate the site of clonal deletion, the D13S25 and D13S319 markers (at 13q14.3) and probes for exons 11 and 26-27 of the BRCA2 gene (at 13q12). Homozygous and heterozygous deletion at the 13q14.3 region was found in five and 17 patients, respectively. Despite the recent report of the BRCA2 gene involvement in >80% of CLL patients, we failed to detect a single case of homozygous or heterozygous deletion involving the 13q12 region. Our data support previous findings that the 13q14.3, and not the 13q12 region, is the major site of candidate tumour suppressor gene(s) in CLL. PMID- 9217188 TI - Photopheresis in paediatric patients with drug-resistant chronic graft-versus host disease. AB - Photopheresis (ECP) is a new type of photochemotherapy, used for the treatment of oncological and autoimmune diseases. Lymphocytes are drawn from the patients by leukapheresis, treated with 8-methoxypsoralen (8-MOP) and ultraviolet light A (UVA) in an extracorporeal system and then reinfused. Skin exposure to 8-MOP and UVA (PUVA) has been shown to relieve cutaneous symptoms of graft-versus-host disease (GVHD) in bone marrow transplant (BMT) recipients. ECP, which is similar in some ways to PUVA, has been used in this study to treat four paediatric patients who developed chronic GVHD following BMT and in whom GVHD had failed to respond to conventional immunosuppressive therapy. Following ECP, skin lesions cleared almost completely and pulmonary function tests improved in two of three patients with cutaneous and lung involvement. Serum bilirubin and transaminases gradually normalized, and gammaGT decreased considerably in the remaining patient who had a severe cholestatic hepatopathy. The Karnofsky performance score increased to 90% in the three patients with positive responses to ECP and remained unchanged (40%) in the patient who did not respond. Immunosuppressive therapy was reduced in three patients and eventually discontinued in two. No significant side-effects were observed during the treatment. Our results suggest that ECP is a non-aggressive treatment that may benefit patients with chronic GVHD who do not respond to standard immunosuppressive therapy. PMID- 9217189 TI - IBMTR Severity Index for grading acute graft-versus-host disease: retrospective comparison with Glucksberg grade. AB - Acute graft-versus-host disease (GVHD) severity is graded by pattern of organ involvement and clinical performance status using a system introduced by Glucksberg and colleagues 21 years ago. We examined how well Glucksberg grade predicted transplant outcome and constructed a Severity Index not requiring subjective assessment of performance in 2881 adults receiving an HLA-identical sibling T-cell-depleted (n = 752) or non-T-cell-depleted (n = 2129) bone marrow transplant for leukaemia between 1986 and 1992. Relative risks (RR) of relapse, treatment-related mortality (TRM) and treatment failure (TF) (relapse or death) were calculated for patients with (Glucksberg Grade I, II or III/IV acute (GVHD) versus those without acute GVHD and for patients with distinct patterns of organ involvement regardless of Glucksberg grade. Using data for non-T-cell-depleted transplants, a Severity Index was developed grouping patients with patterns of organ involvement associated with similar risks of TRM and TF. Higher Glucksberg grade predicted poorer outcome; however, patients with the same grade but different patterns of skin, liver or gut involvement often had significantly different outcomes. The revised Severity Index groups patients in four categories, A-D. Compared to patients without acute GVHD, RRs (95% confidence interval) of TF were 0.85 (0.69, 1.05) for patients with Index A, 1.21 (1.02, 1.43) with B, 2.19 (1.78, 2.71) with C, and 5.69 (4.57, 7.08) with D. Prognostic utility of the Index was tested in patients receiving T-cell-depleted transplants; similar RRs of TF were observed. An acute GVHD Severity Index is proposed to enhance design and interpretation of clinical trials in the current era of allogeneic blood and bone marrow transplantation. PMID- 9217190 TI - Avascular necrosis of bone after allogeneic bone marrow transplantation: analysis of risk factors for 4388 patients by the Societe Francaise de Greffe de Moelle (SFGM). AB - Increasing numbers of patients are surviving after allogeneic bone marrow transplantation and are therefore at risk for developing late complications. Among these complications, avascular necrosis of bone has been reported, but only two single-centre studies included sufficient patients to enable analysis of the risk factors for developing avascular necrosis. In this multicentre retrospective study the aim was to assess risk factors for this complication in a large number of patients. The population consisted of 4388 patients, recorded in the Societe Francaise de Greffe de Moelle (SFGM) data base, who had undergone a single allogeneic bone marrow transplant between January 1974 and December 1993. 77 patients developed avascular necrosis leading to a 4.3% projected incidence at 5 years. Symptoms developed 2-132 months after transplantation. In these 77 patients a mean of 1.87 joints per patient were affected (range 1-7). The hip joint was the most often affected (88% of patients) and 48% of the patients required joint replacement. All but two patients received steroids for acute and/or chronic graft-versus-host disease (GvHD) over a mean period of 15 months. In univariate analysis, among eight factors tested as risk factors for developing avascular necrosis, six were shown to be linked to an increased risk: older age, diagnosis of aplastic anaemia or acute leukaemia, an irradiation-based conditioning regimen, type of GvHD prophylaxis regimens, acute and chronic GvHD. In multivariate logistic regression analysis, five factors remained significantly associated with an increased risk for developing avascular necrosis: chronic GvHD (odds ratio (OR) 3.52), acute GvHD (OR 3.73), age > 16 years (OR 5.81), aplastic anaemia (OR 3.90), and acute leukaemia (OR 1.72). Avascular necrosis is not a rare late complication of allogeneic bone marrow transplantation causing significant morbidity. In this study, older age, initial diagnosis, and GvHD and/or its treatment with steroids emerged as significant risk factors. PMID- 9217191 TI - Multilineage mobilization of peripheral blood progenitor cells in humans following administration of PEG-rHuMGDF. AB - The most important physiological regulator of megakaryocytopoiesis is the ligand for the c-mpl receptor (thrombopoietin/megakaryocyte growth and development factor, MGDF). We examined the effect of pegylated-recombinant human MGDF (PEG rHuMGDF): patients received PEG-rHuMGDF at doses of 0.03, 0.1, 0.3 or 1.0 microg/kg/d or placebo for 10d maximum in a double-blinded randomized study. There was a dose-dependent elevation in circulating platelet counts but no alteration in erythrocyte or total leucocyte counts. The number of bone marrow megakaryocytes was increased approximately 2-fold. The frequency of bone marrow progenitor cells was not altered. In contrast, both to the bone marrow results and to published pre-clinical data, there was a dose-dependent mobilization into the blood of progenitor cells of multiple cell lineages. Increased levels of Meg CFC (maximum increase 30-fold), day 7 and day 14 GM-CFC and BFU-E were demonstrated at doses of 0.3 and 1.0 microg/kg/d PEG-rHuMGDF. At 0.1 microg/kg/d, mobilization of Meg-CFC alone occurred in two-thirds of patients. Maximum blood levels of progenitor cells occurred at day 12. Thus, administration of PEG rHuMGDF to humans resulted in mobilization of progenitor cells of multiple lineages despite its 'lineage-specific' activity on mature cell development. PMID- 9217192 TI - Immunomagnetic selection of CD34+ peripheral blood stem cells for autografting in patients with breast cancer. AB - Contamination of transplants with tumour cells may contribute to relapse after peripheral blood stem cell transplantation (PBSCT). We studied the feasibility of CD34+ cell selection from blood-derived autografts obtained following G-CSF supported cytotoxic chemotherapy in a group of 25 patients with breast cancer (10 with high-risk stage II/III and 15 with stage IV without bone or bone marrow involvement). Using immunomagnetic beads (Isolex 300 SA. Baxter) CD34+ cells were enriched and released by chymopapain resulting in a median purity of 95% (range 82-99%) and a median recovery of 80% (range 27-132%). The enrichment procedure did not change the proportion of CD34+ subsets coexpressing HLA-DR, CD38 and Thy 1, while L-selectin was removed from the cell surface following selection. Using a sensitive immunocytological technique with a cocktail of epithelial-specific antibodies (anti-cytokeratin 8, 18 and 19; HEA125; BM7 and BM8), five leukaphereses products contained epithelial cells, whereas the selected CD34+ cell fraction was free of tumour cells. A neutrophil count of 0.5 x 10(9)/l and a platelet count of 20 x 10(9)/l was reached after a median time of 14 and 10d following 40 high-dose chemotherapy (HDC) cycles. Our results indicate that immunomagnetic selection of CD34+ cells yields highly purified autografts devoid of tumour cells whereas the engraftment ability of the progenitor and stem cells is fully retained. PMID- 9217193 TI - Variations in culture pH affect the cloning efficiency and differentiation of progenitor cells in ex vivo haemopoiesis. AB - Haemopoietic cultures may experience pH variations of as much as 0.5 units depending on culture duration and cell density. Since pH is a potent modulator of cellular proliferation and differentiation, we examined its effects on the performance of both semisolid and liquid haemopoietic cultures. Culture pH was found to have substantial effects both on progenitor cloning efficiency (as measured in liquid cultures) and on progenitor cell differentiation (as measured in methylcellulose cultures). Liquid cultures were conducted with both peripheral blood (PB) mononuclear cells (MNCs) and cord blood (CB) MNCs using growth factor combinations that promote either erythroid expansion (IL-3/IL-6/SCF/Epo) or granulocyte/macrophage expansion (IL-3/IL-6/SCF/G-CSF/GM-CSF). Reduced pH was found to have either a positive or neutral effect on the expansion and cloning efficiency of progenitors in ex vivo liquid cultures. Cloning efficiencies of PB BFU-E in the erythroid combination were 9-fold higher at low pH (7.1) when compared to high pH (7.6). A small pH increase of 0.2 units over physiological values consistently produced significant reductions (42-85%) in cloning efficiencies for all cell types and cytokine combinations tested. Methylcellulose cultures conducted using CB MNC and PB MNC indicated that differentiation of CFU GM into progeny was optimal between pH 7.2 and 7.4. The differentiation of erythroid progenitors (BFU-E) progressively increased as pH was increased from 6.95 (no colonies detected) to 7.4 (maximum colonies detected), to 7.6 (maximum haemoglobin content). Methylcellulose cultures using PB CD34+ cells exhibited similar patterns to the MNC cultures. We conclude that even small variations in pH substantially affected the performance of human haemopoietic cultures. The erythroid lineage was particularly sensitive, with its extent of differentiation increasing with increasing pH. PB progenitors are more sensitive to pH variations than CB progenitors. PMID- 9217194 TI - Treatment of adults with acute lymphoblastic leukaemia in first bone marrow relapse: results of the ALL R-87 protocol. AB - Sixty-one adults aged <55 years with acute lymphoblastic leukaemia (ALL) in first bone marrow relapse were enrolled in an Italian cooperative study (ALL R-87 protocol) from 12 GIMEMA Institutions. The treatment programme consisted of: (1) an induction phase with intermediate-dose cytarabine (IDARA-C 1 g/m2, 6 h daily infusion x 6 d) plus idarubicin (IDA; 5 mg/m2/d x 6 d) and prednisone (40 mg/m2/d x 21 d), (2) a consolidation phase followed by (3) bone marrow transplant (BMT). Median first complete remission (CR) duration was 8.5 months (range 1-54 months). 34/61 patients achieved CR (56%); 24 (39%) failed to respond and three (5%) died during induction. Most responders (24 patients) could not enter the BMT programme; 15 relapsed early (median time to relapse 2 months); nine were withdrawn due to toxicity and one died in CR of infection. Nine of the 34 CRs underwent BMT (five autologous and four allogeneic). Three of the four allotransplanted patients are alive in continuous CR at 22, 43 and 63 months; only one of the five who underwent an autologous BMT is alive in CR at 46 months. The estimated disease-free survival (DFS +/- SE) at 36 months was 0.16 +/- 0.08 for all responders. Univariate analysis showed that previous therapy was the only prognostic factor influencing DFS. The estimated probabilities of event-free survival (EFS +/- SE) and survival +/- SE at 37 months were 0.09 +/- 0.04 and 0.10 +/- 0.04, respectively. The EFS was significantly better in patients with a preceding CR > or = 24 months, compared to those with a shorter first remission. Our results confirm the tolerance and efficacy of IDARA-C plus IDA in inducing CR in poor-risk adult ALL. Even though the number of transplanted patients was small, allogeneic BMT seems to give a real opportunity of cure in this category of patients. PMID- 9217195 TI - Prevalence and clinical significance of hepatitis G virus infection in adult beta thalassaemia major patients. AB - The risk of polytransfused patients for hepatitis C virus (HCV) infection is likely to extend to another recently identified member of the Flaviviridae, hepatitis G virus (HGV). We investigated the prevalence of HGV in 40 adult Italian patients with transfusion-dependent thalassaemia and evaluated the clinical significance of HGV infection. HGV-RNA was detected in 9/40 patients (22.5%). HGV infection was significantly associated with HCV viraemia (P = 0.0012), with all patients positive for HGV being also viraemic for HCV. Overall, the clinical picture of patients with HCV/HGV co-infection was not different from that of patients with isolated HCV. However, patients co-infected with both viruses had lower values of alanine-transferase (P = 0.035) and a lower titre of HCV viraemia (P = 0.042) in the absence of other evident factors which could influence the clinical expression of HCV infection. In conclusion, HGV is highly prevalent among Italian polytransfused patients. No evidence of a clinically significant pathogenic role for HGV in liver disease could be found in these patients. In a subset of cases a possible interference of HGV with HCV infection was observed. PMID- 9217196 TI - Types and sources of fuels for platelets in a medium containing minimal added fuels and a low carryover plasma. AB - The storage of platelets in synthetic media can result in plasma savings and reduced transfusion reactions. Accordingly, a wide range of storage formulations have been developed with the aim of replacing at least a proportion of the plasma in the storage medium. However, the concentrations and types of fuels in the carryover plasma, and the utilization of these fuels by platelets in storage, has not been investigated. We have developed a system which can measure total ATP turnover, and the contribution to total ATP turnover by the oxidation of various fuels and by lactate production, in a bag of partially purified platelets in a buffered saline with minimal carryover citrate phosphate double dextrose (CP2D) plasma. Carryover plasma was about 1% and the final platelet suspension contained, on average, 0.62 mM glucose, 9.6 mg/l free fatty acids, 32 mg/l triglycerides and 0.23 mM total amino acids. The oxidation of carbohydrate (glucose, glycogen and lactate) accounted for 60% of total ATP turnover. The platelets also produced lactate (<6% of total ATP turnover) and consumed free fatty acids and amino acids/proteins (15.2% of total ATP turnover). Therefore we have identified the fuels that account for about 80% of oxygen consumption and ATP turnover by platelets in a medium with low carryover plasma. The implications of these data for storage strategies are discussed. PMID- 9217197 TI - Is antenatal antibody screening worthwhile in Chinese? AB - A total of 1997 pregnant women were screened during their first antenatal visit for irregular antibodies for the prevention of haemolytic disease of the newborn. Patient sera were tested against a panel of group O screen cells including one with the expression of Miltenberger determinants GP.Mur. 17 women (0.85%) had irregular antibodies of which four were of potential clinical significance, including one with anti-D, two with anti-E and one with anti-D, anti-E and anti G. Although antenatal antibody screening is mandatory in Western populations, our results suggest that this may not be necessary in the Chinese population except for those who are Rh D-negative or who have a history of haemolytic disease of the newborn. PMID- 9217199 TI - Royal College of Physicians of Edinburgh/Royal College of Obstetricians and Gynaecologists Consensus Conference on anti-D prophylaxis, 7-8 April 1997. PMID- 9217198 TI - Clonal haemopoiesis in normal elderly women: implications for the myeloproliferative disorders and myelodysplastic syndromes. AB - Studies of X chromosome inactivation patterns are central to many aspects of our understanding of the pathogenesis of haematological malignancies. In patients with myeloproliferative disorders and myelodysplastic syndromes the demonstration of skewed X inactivation patterns in multiple haemopoietic lineages has been taken to indicate a stem cell origin for these groups of diseases. However, stem cell depletion or selection pressures can also produce skewed X inactivation patterns and might increase with age. We have therefore used the HUMARA assay to study X inactivation patterns of elderly patients with myeloproliferative disorders together with an age-matched control group of normal elderly women. A clonal pattern (clonal granulocytes and polyclonal T cells) was observed in 23.1% of normal women and 63.4% of patients with myeloproliferative disorders. This is the first report of X inactivation patterns in purified subpopulations of blood cells in normal elderly women. These results have three significant implications. Firstly, the finding of clonal granulocytes and polyclonal T cells in normal elderly women is likely to reflect age-related stem cell depletion or selection pressures. Secondly, the demonstration of clonal granulocytes and polyclonal T cells is not a useful diagnostic marker for myeloproliferative disorders or myelodysplastic syndromes in elderly women. Thirdly, our data raise the possibility that clonal blood cell patterns may precede rather than follow mutations which subsequently give rise to myelodysplastic or myeloproliferative phenotypes. PMID- 9217200 TI - Combination of interferon-alpha and hydroxyurea in the treatment of idiopathic hypereosinophilic syndrome. PMID- 9217201 TI - Provision of platelets for severe neonatal alloimmune thrombocytopenia. PMID- 9217202 TI - Ongoing somatic mutation in mantle cell lymphomas questioned. PMID- 9217203 TI - Cytokine-receptors repertoire in a pre-osteoclast cell line. PMID- 9217204 TI - Effect of sample storage on the International Normalized Ratio. PMID- 9217205 TI - Incidence of Ph1 positive ALL in unselected, population-based study of adult ALL. PMID- 9217206 TI - Tests for heparin-induced thrombocytopenia in primary antiphospholipid syndrome. PMID- 9217207 TI - Factor V Leiden: prevalence in the indigenous population and cases of thrombosis in North India. PMID- 9217208 TI - Earliest evidence for arthrogryposis multiplex congenita or Larsen syndrome? AB - A sixteenth-century illustrated pamphlet from Great Britain suggests that documentary evidence may permit accurate diagnosis of pathological conditions in earlier societies. The document is of particular importance, since the presented congenital abnormalities, including cleft lip, spina bifida cystica, genu recurvatum, and talipes deformity are reported rarely in archaeological skeletal material. It is suggested that the combination of abnormalities may represent the earliest case of arthrogryposis multiplex congenita or Larsen syndrome. PMID- 9217209 TI - Rapid identification of marker chromosomes using primed in situ labeling (PRINS). AB - Primed in situ labeling (PRINS) is a relatively new technology with wide-ranging applications in clinical cytogenetics. Using PRINS, we have identified the chromosomal origin of marker chromosomes in three patients. In the first patient with primary amenorrhea, we were able to confirm the marker chromosome as originating from an X. In the second (prenatal) case, PRINS allowed us to determine rapidly the origin of the marker as a Y chromosome. In the third patient with minor anomalies, the marker was identified as derived from a chromosome 18. In all three cases, application of PRINS permitted us to characterize the marker chromosomes within 1 hour after the slides were prepared. The methodology is simple, has added advantages over conventional fluorescence in situ hybridization (FISH), and can be used as a viable and effective alternative to FISH in clinical cytogenetic diagnosis. PMID- 9217210 TI - Gillespie syndrome: a report of two further cases. AB - We describe two unrelated patients with Gillespie syndrome (partial aniridia, cerebellar ataxia, and mental retardation). The typical presentation is the discovery of fixed dilated pupils in a hypotonic infant. The iris abnormality is specific and seems pathognomonic of Gillespie syndrome. It can be distinguished clinically from other forms of aniridia and a presumptive diagnosis of Gillespie syndrome can be made in the first months of life on the basis of the ocular findings. Neurological involvement includes marked motor delay, hypotonia, disabling ataxia, and usually mental retardation. Cerebral and cerebellar atrophy with white matter changes on MRI scan were present in our second patient suggesting that patients with Gillespie syndrome may have more extensive CNS involvement than previously described. The parents of this child were first cousins; thus Gillespie syndrome may be heterogeneous with autosomal recessive and autosomal dominant forms. PMID- 9217211 TI - Partial 9p monosomy in a girl with a tdic(9p23;13p11) translocation, minor anomalies, obesity, and mental retardation. AB - We report on a case with a partial monosomy for the regions 9p23 --> pter and 13p11 --> pter as a result of a de novo translocation (9p23;13p11). The patient, a 16-year-old girl, has mental deficiency, obesity, and minor anomalies, including trigonocephaly, hypertelorism and a short, broad neck. Cytogenetic and microsatellite marker analysis allowed us to assign the breakpoint to the chromosomal region 9p23, flanked by the markers D9S144 and D9S157. In an attempt to establish a phenotype-genotype correlation, the clinical manifestations present in our patient are compared to those with partial 9p monosomy and breakpoint in p23, referred to in the literature. PMID- 9217212 TI - A newly recognized autosomal dominant ectodermal dysplasia syndrome: the odonto tricho-ungual-digital-palmar syndrome. AB - We report on two brothers, their mother, and 18 other relatives of five generations presenting an apparent newly recognized syndrome involving natal teeth, trichodystrophy, prominent interdigital folds, simian-like hands with transverse palmar creases, and ungual digital dystrophy, inherited as an autosomal dominant trait. In addition, the patients had hypoplasia of the first metacarpal and metatarsal bones and distal phalanges of the toes. PMID- 9217213 TI - Autosomal dominant microcephaly with normal intelligence, short palpebral fissures, and digital anomalies. AB - We describe a family segregating an autosomal dominant mutation producing a syndrome comprising microcephaly with normal intelligence and short palpebral fissures together with variable signs including thumb hypoplasia, shortness of the middle phalanges of the second and fifth fingers, small feet, a gap between the first and second toes, and mild syndactyly of the toes or fingers. A characteristic radiologic finding in our family is thinning of the proximal end of the first metacarpal and shortening of that metacarpal. The severity of these findings was asymmetric in our patients. This syndrome is similar to patients described by Brunner and Winter [1991: J Med Genet 28: 389-394], Feingold [1975: Synd Ident 3:16-17, 1978: Hosp Prac 13:44-49], and Konig et al. [1990: Dysmorphol Clin Genet 4:83-86]. PMID- 9217214 TI - Correlation between the incidence of myotonic dystrophy in different groups in Israel and the number of CTG trinucleotide repeats in the myotonin gene. AB - Myotonic dystrophy (DM) is associated with an increased number of CTG repeats in the 3' untranslated region of the myotonin gene. Because DM has been observed less frequently in Ashkenazic Jews and non-Jews than in North African and Yemenite Jews in Israel, a study of the CTG repeat polymorphism was undertaken in these four groups. Alleles from 126 unrelated healthy North African Jews, 103 Yemenite Jews, 103 Ashkenazic Jews, and 106 Israeli Moslem Arabs were studied by PCR analysis of the trinucleotide repeat in the DM gene, and the size distribution of the CTG repeat was determined. The alleles ranged in length from 5-28 repeats in the Yemenite Jews, 5-26 in Muslim Arabs and North African Jews, and 5-23 in the Ashkenazic Jews. North African and Yemenite Jews were found to have significantly more large repeats in the normal range than Ashkenazic Jews and Muslim Arabs (for over 18 repeats: 9.1% and 13%, respectively, compared to 2.4% and 3.3%, respectively; P < 0.0001). It is suggested that the more frequent occurrence of large CTG repeats in the normal range may represent a greater predisposition to DM. PMID- 9217215 TI - Maternal balanced translocation leading to partial duplication of 4q and partial deletion of 1p in a son: cytogenetic and FISH studies using band-specific painting probes generated by chromosome microdissection. AB - A 9-month-old boy with pre- and post-natal growth retardation, microcephaly, plagiocephaly, and several minor anomalies had the initial karyotype: 46,XY,der(1)t(1;?) (p36.1;?). Further analysis showed that the der(1) was derived from an unfavorable segregation of a maternal complex chromosome rearrangement, i.e., 46,XX,der(1)t(1;?) (p36.1;?), der(4)t(4;?)(q?;?). Whole chromosome fluorescence in situ hybridization (FISH) and chromosome microdissection were used to clarify the maternal karyotype as: 46,XX,der(1)t(1;4)(4qter- >4q33::1p36.13-->1qter),der( 4)t(1;4)inv(4)(4pter-->4q31.3::1p36.33- >1p36.13::4q33 -->4q31.3::1p36.33-->1pter). Therefore, the karyotype of the boy actually was 46,XY,der(1)t(1;4) (p36.13;q33). Clinical comparison of the patient's clinical findings showed similarities to individuals with partial del(1p) and dup(4q). To our knowledge the above cytogenetic abnormalities have not been described previously. This case further demonstrates the advantages of chromosome microdissection and FISH in the identification of anomalous chromosome regions and breakpoints. PMID- 9217216 TI - Multiple, juxtasutural, cranial hyperostoses and cardiac tumor: a new hamartomatous syndrome? AB - We report on a Japanese girl with multiple cranial hyperostoses and a cardiac tumor, both of which manifested in early childhood. Unique juxtasutural lesions characterized the cranial findings, including a chain of almost symmetrical osseous protuberances involving the frontozygomatic and frontoparietal junctions, and discrete bony bumps on the right occipitoparietal junction and left temporo occipital junction. These lesions histologically consisted of thickened mature bone intervened with sparse fibrous tissues, mimicking osteoma. The cardiac mass remained pathologically unknown, but was shown to have fatty elements on magnetic resonance imaging (MRI). The patient showed no evidence of gnathic hyperostoses, ophthalmological abnormalities, skin lesions, or other visceral abnormalities, which ultimately precluded known hamartomatous syndromes with craniofacial hyperostoses, such as Gardner and Proteus syndromes. Yet regional Proteus syndrome could not be completely excluded. The craniofacial deformity as a sequel of hyperostoses in our patient superficially resembled that of X-linked calvarial hyperostosis; however, the vacuolated histiocytes that characterized the hyperostotic lesions were not found in our patient. The present disorder may represent a hitherto unknown hamartomatous syndrome. PMID- 9217217 TI - Identification of two novel and four uncommon missense mutations among chinese Gaucher disease patients. AB - Gaucher disease is the most prevalent lysosomal storage disease. It is panethnic and results from an inherited deficiency of glucocerebrosidase. Most mutations to date have been identified among Jewish and non-Jewish Caucasian patients; mutations in Chinese patients are largely unknown. We have performed nucleotide sequence analysis of PCR-amplified glucocerebrosidase genomic DNA from five unrelated Chinese patients affected with type 1 (non-neuropathic) Gaucher disease. A novel heterozygous C --> T mutation at cDNA nucleotide position 475 (R120W) was detected in a patient who is also heterozygous for a C --> T transition at cDNA nucleotide position 259 (R48W). In a second patient, a novel, heterozygous T --> G transversion at cDNA 226 (F37V) was detected. Mutation 1448 (L444P), the most prevalent mutation among non-Jewish Caucasian Gaucher patients, was found in the heterozygous form in four patients. The mutations in the second Gaucher allele in the other three patients are mutations 254 (G46E), 680 (N188S), and 754 (F213I), which were recently reported in Korean, Arab, and Chinese (Taiwanese) patients. We have developed screening methods that utilize PCR amplification of glucocerebrosidase genomic DNA and Eco571, Nci1, Hinc11, BsaJ1, and Bsr1 restriction endonuclease analyses for the detection of each of these mutations. The population genetics of some of these Gaucher alleles and their implications in genotype/phenotype correlation are discussed. PMID- 9217218 TI - Restrictive dermopathy: report and review. AB - Restrictive dermopathy (RD) is a lethal autosomal recessive genodermatosis (MIM No. 275210) in which tautness of the skin causes fetal akinesia or hypokinesia deformation sequence (FADS). Polyhydramnios with reduced fetal movements is followed by premature delivery at around 31 weeks gestation. Manifestations include a tightly adherent, thin, translucent skin with prominent vessels, typical facial changes, generalized joint contractures, enlarged fontanelles, dysplasia of clavicles, respiratory insufficiency, and an enlarged placenta with short umbilical cord. Histologic abnormalities of the skin include thin dermis with paucity and hypoplasia of the appendages and abnormally arranged collagen bundles. Elastic fibers are nearly missing. The subcutaneous fat is slightly increased. These skin findings usually appear after 22 or 24 weeks of gestation, which is why prenatal diagnosis with skin biopsy may fail. This disease is easily differentiated from other congenital FADS, such as Pena-Shokeir syndrome, COFS syndrome, Parana hard-skin syndrome, etc. We report on an affected boy of consanguineous parents and 30 previous cases are reviewed. PMID- 9217219 TI - Schopf-Schulz-Passarge syndrome with an unusual pattern of inheritance. AB - Schopf-Schulz-Passarge syndrome is a rare form of ectodermal dysplasia comprising hypotrichosis, hypodontia, unusual eyelid cysts, palmar-plantar keratosis, and nail dystrophy. To date, ten cases have been reported; all except one are compatible with autosomal recessive inheritance. We report on a family in which three full sibs and one half-sib have Schopf-Schulz-Passarge syndrome, yet there is no other evidence of dominant transmission in prior or subsequent generations. Possible explanations are discussed. PMID- 9217220 TI - Severe clinical phenotype due to an interstitial deletion of the short arm of chromosome 1: a brief review. AB - We report on a newborn girl with malformed ears, bilateral cleft lip and cleft palate, complex congenital heart disease, absent left thumb, and rib abnormalities. Cytogenetic analysis demonstrated a de novo interstitial deletion of the short arm of chromosome 1 [46,XX,del(1)(p21p22.3)]. Reports of interstitial deletions on the short arm of chromosome 1 are rare. However, when comparing this patient's phenotype to others with deletions of 1p, we found that the current case was much more severely affected than previously reported cases. PMID- 9217221 TI - A further case of vertical transmission of proximal femoral focal deficiency? AB - We report on a newborn boy with congenital asymmetrically hypoplastic fibulae, lateral oligodactyly, and mild left ectrodactyly. The patient's grandfather had a short femoral shaft with a slightly smaller collodiaphyseal angle on the left as compared to the right side, probably a proximal focal femoral deficiency (PFFD). The upper limbs were not affected in either patient. PFFD in the grandfather and hypoplastic fibulae with lateral ray defects in the grandson raise the possibility of genetic transmission, specifically autosomal-dominant inheritance with variable penetrance and expressivity. This case gives further support to the fibular developmental field concept postulated by Lewin und Opitz [1986: Am J Med Genet (Suppl) 2:215-238]. PMID- 9217222 TI - Dilemmas of anonymous predictive testing for Huntington disease: privacy vs. optimal care. AB - Some persons at risk for Huntington disease (HD) seek predictive testing under the protection of anonymity to reduce the risk of insurance discrimination for themselves and their families. While Canadian and European health care systems seem to limit insurance discrimination to life and disability insurance, U.S. residents do not have national health insurance and are concerned about health insurance discrimination. Two persons residing outside Canada requested predictive testing anonymously. Their primary reason for doing so was to avoid the risks of medical insurance discrimination. After a detailed preparatory session and agreement to counselling and to receipt of results in person, we agreed to provide anonymous testing to these persons. One participant, whose psychological assessment was unremarkable, coped well with the predictive testing process and did not have the CAG expansion. The other participant had considerable emotional problems prior to testing, which necesitated postponement of discussion of results and referral for psychiatric assessment and support. Both participants had difficulty maintaining anonymity. The provision of anonymous predictive testing raises several problems. With anonymous testing, clinicians cooperate with participants to exclude insurance companies from information. This may invalidate the contract with insurance companies. A policy response by insurance companies or a universal health care system to protect individuals is preferable. Individuals who request anonymous testing may be precisely those most vulnerable and in need of additional support and counselling. However, the preservation of anonymity is a burden to participants and may frustrate the clinicians' ability to establish rapport in counselling and to provide appropriate follow-up typically available through genetic counselling in predictive testing programs. PMID- 9217223 TI - Neonatal severe hyperparathyroidism, secondary hyperparathyroidism, and familial hypocalciuric hypercalcemia: multiple different phenotypes associated with an inactivating Alu insertion mutation of the calcium-sensing receptor gene. AB - Neonatal severe hyperparathyroidism (NSHPT) is considered an autosomal-recessive disorder, attributable in many cases to homozygous inactivating mutations of the Ca++-sensing receptor (CASR) gene at 3q13.3-21. Most heterozygotes are clinically asymptomatic but manifest as familial (benign) hypocalciuric hypercalcemia (FHH) with a laboratory profile that is variably and sometimes only marginally different from normal. In 5 NSHPT cases from 3 Nova Scotian families, we found homoallelic homozygosity for an insertion mutation in exon 7 of CASR that includes an Alu repeat element with an exceptionally long polyA tract. Four of the 5 NSHPT infants were treated by parathyroidectomy more than a decade ago and are well now. A fifth went undiagnosed until adulthood and has profound musculoskeletal and neurobehavioral deficits. Among 36 identified FHH heterozygotes are 3 individuals with an unexpected degree of hypercalcemia and elevated circulating parathyroid hormone levels consistent with secondary hyperparathyroidism. Two are obligately heterozygous offspring of NSHPT mothers with surgical hypoparathyroidism and variable compliance with vitamin D therapy. The other is an adult with coexistent celiac disease in whom hyperparathyroidism, probably secondary to vitamin D deficiency, led to surgery. In counseling affected families, the heterozygous state should not be considered entirely benign, since FHH heterozygotes, particularly infants, may be prone to secondary hyperparathyroidism and symptomatic hypercalcemia. In such families, molecular diagnosis will allow for unambiguous identification of at-risk individuals. PMID- 9217224 TI - Expansion of the phenotype in Hennekam syndrome: a case with new manifestations. AB - We report on a female with lymphedema, facial anomalies, intestinal lymphangiectasia, and moderate mental retardation consistent with the diagnosis of Hennekam syndrome. In addition, she had a number of other anomalies not previously described in this autosomal recessive disorder, including a congenital heart defect, atretic ear canals, vesicoureteral reflux, and rectal prolapse. PMID- 9217225 TI - Frequency of associated anomalies in congenital hypoplasia of depressor anguli oris muscle: a study of 50 patients. AB - Aside from congenital heart disease, anomalies associated with unilateral hypoplasia of the depressor anguli oris muscle have not been well-documented in large series. We evaluated the associated anomalies in 50 infants or children with this disorder (male:female = 2:1) and found accompanying anomalies in 35 (70%) of 50 cases. They included anomalies of the head and neck (48%), heart (44%), skeleton (22%), genitourinary tract (24%), central nervous system (10%), gastrointestinal tract (6%), and miscellaneous minor anomalies (8%). Nearly half of our cases (22/50) had at least 2 associated systemic anomalies. Failure to thrive and psychomotor retardation were found in 5 (10%) and 3 (6%) patients, respectively, on follow-up. Three infants died neonatally of severe heart disorders, and the other one died of central nervous system anomalies. The above findings indicate that a thorough search for associated anomalies, particularly in the cardiovascular system, should be performed in all newborns with asymmetric crying face. PMID- 9217226 TI - Cervical ribs in fetuses with Ullrich-Turner syndrome. AB - The purpose of this study was to analyze the cervical skeleton in fetuses with Ullrich-Turner syndrome (45,X) in a search for skeletal characteristics in the neck region affected by hygroma. In connection with requested autopsies, 9 second trimester human fetuses were investigated radiographically by whole-body and special radiography of the spine. The presence of unilateral or bilateral cervical ribs was a constant finding which seems applicable as a phenotypic characteristic of Ullrich-Turner syndrome. This study may also be important in the diagnosis of newborn infants with Ullrich-Turner syndrome. PMID- 9217228 TI - Trichorrhexis nodosa and lip pits in autosomal dominant ectodermal dysplasia- central nervous system malformation syndrome. AB - A Dandy-Walker-like malformation was observed in a retarded girl who had signs of hidrotic ectodermal dysplasia. This is the third report of the rare triad ectodermal dysplasia-CNS malformation-mental retardation. We observed additional findings, such as submucous cleft palate with lip pits and trichorrhexis nodosa. The proposita's mother had similar hair and facial changes. Two maternal relatives had cleft palate. Autosomal dominant inheritance is suggested. PMID- 9217227 TI - Pseudodiastrophic dysplasia type Burgio in a newborn. AB - Pseudodiastrophic dysplasia is a distinct disorder that differs from diastrophic dysplasia on the basis of elbow and proximal interphalangeal joint dislocations, platyspondyly, and scoliosis. We report on a new patient with this rare skeletal dysplasia and two previously undescribed major malformations: omphalocele and complex heart defect. PMID- 9217229 TI - Familial patent ductus arteriosus: a further case of CHAR syndrome. AB - We report on a family with patent ductus arteriosus, a distinctive facial appearance with eyebrow flare, a short nose and "duck-bill lips," polydactyly, and fifth finger clinodactyly. The facial traits were consistent with CHAR syndrome. We provide further evidence for evolution of the phenotype with age and describe the previously unreported finding of interstitial polydactyly in this syndrome. PMID- 9217230 TI - Short stature, Robin sequence, cleft mandible, pre/postaxial hand anomalies, and clubfoot: another affected Brazilian patient born to consanguineous parents. PMID- 9217231 TI - Cryptic pericentric inversion of chromosome 17 detected by fluorescence in situ hybridization study in familial Miller-Dieker syndrome. PMID- 9217232 TI - Questions and problems in direct predictive testing for Huntington's disease. PMID- 9217233 TI - Major factors determining the frequencies of hemoglobinopathies in Oman. PMID- 9217234 TI - Pfeiffer mutation in an Apert patient: how wide is the spectrum of variability due to mutations in the FGFR2 gene? PMID- 9217235 TI - Novel mutation of the myelin Po gene in a pedigree with Charcot-Marie-Tooth disease type 1B. PMID- 9217236 TI - Unsuspected mutation in a family with congenital adrenal hyperplasia. PMID- 9217237 TI - Pro-oxidant effects of delta-aminolevulinic acid (delta-ALA) on Chinese hamster ovary (CHO) cells. AB - delta-Aminolevulinic Acid (delta-ALA) is a heme precursor accumulated in lead poisoning and acute intermittent porphyria. Although no single mechanism for lead toxicity has yet been defined, recent studies suggest at least some of the lead induced damage may originate from delta-ALA-induced oxidative stress. The present study was designed to test the hypothesis that delta-ALA accumulation in Chinese hamster ovary (CHO) cells contributes to the cumulative oxidative challenge of lead poisoning as indicated by the oxidative stress parameters glutathione (GSH), glutathione disulfide (GSSG), malondialdehyde equivalents (MDA), and catalase (CAT). It will also examine the possibility that this oxidative challenge can be reversed by treatment with an antioxidant such as N-acetylcysteine (NAC). First in vitro administration of delta-ALA to CHO cells was found to have a concentration-dependent inhibitory effect on colony formation and cell survival. NAC administration was shown to alleviate this inhibition in CHO survival. The oxidative status of CHO cell cultures exposed to increasing concentrations of delta-ALA was then examined. Decreases in GSH levels (P < 0.05) were observed in the delta-ALA-treated cultures as compared to the controls, while GSSG and MDA levels were significantly increased in delta-ALA-treated cells (P < 0.05). CAT activity was not significantly affected. NAC administration concurrent with delta ALA exposure resulted in GSH and GSSG levels similar to the control levels, while no significant improvement in MDA was observed. These results indicate a state of oxidative stress and suggest that the delta-ALA- induced inhibitory effect on CHO colony formation may be due to its pro-oxidant effect. To assess whether this oxidative challenge would induce antioxidant increases during extended exposure to delta-ALA, CHO cells were exposed to 5 mM delta-ALA for increasing time periods. The GSH and GSSG levels were measured and a rebound effect was observed after 12 h of delta-ALA exposure. PMID- 9217238 TI - The involvement of metallothionein in the intestinal absorption of cadmium in mice. AB - Female mice were exposed to a single dose of 0.005 to 5 microg Cd/kg body wt., in order to test the hypothesis that once the Cd-binding capacity of intestinal metallothionein is saturated. Cd becomes more readily available for transfer from the mucosa to the circulatory system, causing an increase in Cd absorption. In this case the binding of Cd to MT would act as a barrier against Cd absorption, thus protecting the organism from accumulation and toxic effects of Cd in target organs such as the kidney. In mice the fractional Cd uptake (% of dose) in the duodenum, which was the main site of Cd uptake in the intestine, was not influenced by the Cd dose 6 h after dosage. However, the percentage of cytosolic Cd associated to MT in the duodenum decreased when the Cd dosage increased from 0.005 or 0.025 microg/kg to 5 microg/kg. Concomitantly, the percentage bound to low-molecular-weight (LMW) ligands increased, indicating saturation of the Cd binding capacity of MT. Nevertheless, the fractional absorption was not dosage dependent in the dosage interval studied. Moreover, there was no statistically significant correlation between the percentage of cytosolic Cd bound to MT and the percentage of Cd absorbed. Thus, our results do not support the hypothesis that the intestinal Cd absorption is increased when the Cd-binding capacity of intestinal MT is saturated. PMID- 9217239 TI - Sodium lauryl sulfate and triclosan: in vitro cytotoxicity studies with gingival cells. AB - Triclosan and sodium lauryl sulfate (SLS) are antimicrobial agents used, both singularly and in combination, in dentifrices and mouth-rinses. Studies by Waaler et al. (Scand. J. Dent. Res. 101 (1993) 192-195) with human volunteers showed that the adverse side-effects induced by SLS in mouth-rinses, i.e. desquamation of oral epithelium and a burning sensation, were lessened by the addition of triclosan. However, Baert et al. (Int. J. Exp. Pathol. 77 (1996) 73-78) showed that triclosan did not protect the hamster cheek pouch mucosa from irritation caused by SLS. The studies presented herein further evaluated, using a cell culture system, the triclosan-SLS interaction. The in vitro cytotoxicities of triclosan and SLS, alone and in combination, were determined with human gingival S-G epithelial cells and GF fibroblasts. The 24-h midpoint (NR50) cytotoxicity values towards the S-G cells were 0.052 mM triclosan and 0.0075% SLS and for the GF fibroblasts the respective values were 0.095 mM triclosan and 0.0127% SLS. Both agents at their NR50 values induced vacuolization. Coexposures of triclosan and SLS were additive in their cytotoxicities towards the S-G epithelial cells and GF fibroblasts. Pretreatment with triclosan potentiated the toxicity of a subsequent exposure of SLS to the S-G cells; a similar pretreatment of the GF fibroblasts with triclosan had no effect on a subsequent challenge with SLS. PMID- 9217240 TI - Vehicle-dependent oral absorption and target tissue dosimetry of chloroform in male rats and female mice. AB - Chloroform-induced toxicity in rodents depends on oral dose regimen. We evaluated the absorption and tissue dosimetry of chloroform after gavage administration in various vehicles to male Fischer 344 rats and female B6C3F1 mice. Animals received a single dose of chloroform in corn oil, water, or aqueous 2% emulphor at doses (15-180 and 70-477 mg/kg for rats and mice) and dose volumes (2 and 10 ml/kg for rats and mice) used in previously reported toxicity studies. Blood, liver, and kidney chloroform concentration-time courses were determined. Gavage vehicle had minimal effects on chloroform dosimetry in rats. In mice, however, tissue chloroform concentrations were consistently greater for aqueous versus corn oil vehicle. At the low dose volume used for rats (2 ml/kg) gavage vehicle may not play a significant role in chloroform absorption and tissue dosimetry, at the higher dose volume used for mice (10 ml/kg), vehicle may be a critical factor. PMID- 9217241 TI - Effect of the route of benzo[a]pyrene administration on sister chromatid exchange and DNA binding in bone marrow of mice differing with respect to cytochrome P450 1A1 induction. AB - The effects of the route of benzo[a]pyrene administration on sister chromatid exchange (SCE) and B[a]P diol epoxide (B[a]PDE)-DNA adducts formation in bone marrow cells of Ah responsive (C57BL/6; B6) and Ah non-responsive (DBA/2; D2) mice were determined. Animals were treated intraperitoneally (i.p.), intragastrically (i.g.) or topically with two 100 mg/kg doses of benzo[a]pyrene 24 h apart and killed 96 h after the first treatment. Significant increase in the frequencies of SCE and the level of B[a]PDE-DNA adducts as measured by synchronous fluorescence spectrophotometry were detected in D2 mice as compared to B6 mice. The route of administration had little effect on SCE levels in bone marrow cells in D2 mice. In B6 mice higher levels of SCE were observed following i.p. administration as compared to i.g. or topical administration. In both strains the highest level of B[a]PDE-DNA adducts was formed after i.p. administration of B[a]P. We conclude that the i.p. route of B[a]P administration is the most effective in inducing SCE and B[a]P-DNA adducts formation. SCE induction does not correlate linearly with the amount of B[a]PDE-DNA adducts formed in these cells after administration of the above dose of B[a]P. PMID- 9217242 TI - Disruptions of the hypothalamo-pituitary-adrenal axis increase anticancer drug lethality in the rat. AB - We have previously shown that toxicity of the anticancer agent hydroxyurea (HU) in the rat is markedly increased by hypophysectomy or adrenalectomy. In this study, we investigated whether increased toxicity in ablated animals is a unique feature of HU or it is shared with other anticancer agents; the toxic effects of five such drugs have been compared in intact, hypophysectomized (HYX) and adrenalectomized (ADX) rats. Bis-chloroethyl-nitrosourea (BCNU, 5-10 mg/kg), busulfan (0.1-10 mg/kg), cyclophosphamide (25-125 mg/kg), 5-fluorouracil (15-75 mg/kg) and vindesine (0.1-0.5 mg/kg) were given to intact and endocrine-ablated rats, and lethality was recorded over 3 weeks. It was found that mortality was low or absent in intact rats, whereas (with the exception of HYX rats receiving the highest dose of busulfan) it was dramatically increased by both hypophysectomy and adrenalectomy. However, replacement treatments with long acting tetracosactrin and corticosterone to HYX and ADX rats respectively afforded significant protection against BCNU toxicity only. We conclude that the integrity of the hypothalamo-pituitary-adrenal axis is needed to tolerate the toxicity of various anticancer drugs, although complex mechanisms appear to underlie such protective effect. PMID- 9217243 TI - Clastogenicity and mutagenicity of hexavalent chromium in lacZ transgenic mice. AB - The clastogenic and mutagenic effects of the hexavalent chromium compound K2CrO4 in lacZ transgenic mice (Muta Mouse) were investigated. Male Muta mice were administered an intraperitoneal dose of 40 mg/kg of K2CrO4 once on each of 2 consecutive days. The K2CrO4 induced a significant increase in the peripheral blood micronucleated reticulocyte count. Also, K2CrO4 induced a statistically significant increase in mutant frequency in the liver but not in the bone marrow on day 7 after the second treatment. The reason for the failure to increase the mutant frequency in the bone marrow may have been the rapid cell turnover rate there. The mutation induced by K2CrO4 in the bone marrow may have occurred in more differentiated cells than stem cells, and the rapid proliferative activity may have caused a rapid decrease in mutated cells by day 7. Further study with a sampling point earlier than day 7 is needed. The results obtained in the present study indicate that K2CrO4 has clastogenic and mutagenic potential in vivo. PMID- 9217244 TI - Summary of the II International Consensus Symposium on Combined Antiviral Therapy and implications for future therapies. PMID- 9217245 TI - Metabolism of ganciclovir and cidofovir in cells infected with drug-resistant and wild-type strains of murine cytomegalovirus. AB - Murine cytomegalovirus (MCMV) has been used extensively as an animal model for human cytomegalovirus (HCMV). Understanding drug resistance and its treatment in MCMV may lead to more effective treatments of HCMV disease. Most ganciclovir resistant HCMV clinical isolates exhibit a decreased capacity to induce ganciclovir phosphorylation (to its biologically active form) in infected cells. Using an MCMV strain resistant to both ganciclovir and cidofovir, the intracellular metabolism of these drugs was studied to determine if MCMV resistance correlates with decreases in drug phosphorylation. The wild-type (WT) MCMV used for comparison was inhibited in plaque reduction assays, by ganciclovir and cidofovir by 50% at 5.1 and 0.24 microM, respectively; the resistant strain was inhibited at 72 and 2.7 microM, respectively. In uninfected, WT, or resistant virus-infected cells, the extent of metabolism of 10 microM ganciclovir or 1 microM cidofovir to intracellular triphosphorylated species was similar. Phosphorylation and catabolism (following drug removal) rates over time were also similar. Intracellular levels of ganciclovir triphosphate and cidofovir diphosphate increased less than two-fold with increasing multiplicity of virus infection. Because few differences in drug phosphorylation between WT and resistant virus-infected cells were found, virus resistance to ganciclovir and cidofovir apparently is not linked to altered drug phosphorylation. Since the viral DNA polymerase is the antiviral target for these compounds, the resistant MCMV is most likely a DNA polymerase mutant. PMID- 9217246 TI - Safety and efficacy of Virend for topical treatment of genital and anal herpes simplex lesions in patients with AIDS. AB - Virend (SP-303), a new topical antiviral agent with activity against herpesvirus, was evaluated in a multicenter, double-blind, placebo-controlled Phase II study for safety and effectiveness against recurrent genital herpes lesions in patients with AIDS. The primary endpoints of this study were complete healing of lesions and time to healing. Patients had a history of recurrent genital or anogenital herpes with at least one lesion and positive HSV culture at enrollment. Participants received Virend (15% ointment; 24 patients) or matching placebo (21 patients) three times a day for 21 days. Excluding two patients in the Virend group who received an initial treatment but were lost to follow-up, 9 of 22 (41%) patients treated with Virend experienced complete healing of their lesions compared with three (14%) patients in the placebo group (P = 0.053). Viral culture revealed that 50% of Virend-treated patients and 19% of placebo-treated patients became culture-negative during treatment (P = 0.06). Based on these preliminary clinical findings, further evaluation of Virend for topical treatment of genital herpes in patients with AIDS is planned. PMID- 9217248 TI - Bioavailability and metabolism of cidofovir following topical administration to rabbits. AB - The bioavailability and metabolism of the antiviral nucleotide analog cidofovir (HPMPC) were examined in New Zealand white rabbits following topical administration to normal and abraded skin. Male rabbits (four per group) received 14C-cidofovir (100 microCi/kg) intravenously (1 mg/kg) as a solution or topically (2 mg/animal) as a 1% w/w gel containing hydroxyethylcellulose (HEC) with or without propylene glycol (PG). The same PG/HEC formulation was applied topically to an abraded skin site in a fourth group of animals. All radioactivity detected in plasma and skin was accounted for by cidofovir. Plasma concentrations of radioactivity declined multiexponentially following intravenous administration, with a terminal half-life of 5.4 h. For intact skin, the absolute bioavailabilities of the HEC and PG/HEC formulations were 0.2 and 2.1%, respectively. For abraded skin, the bioavailability for the PG/HEC gel was 41%. Radioactivity in kidneys was attributed to cidofovir ( > 95%) and cyclic HPMPC. Concentrations in kidney following topical administration of cidofovir to normal skin were < 4% of those following intravenous dosing. Topical application of cidofovir to intact skin led to negligible systemic exposure to the drug. The topical bioavailability and hence the flux of cidofovir through intact skin was enhanced by the presence of PG in the formulation. Abrasion of the skin removed the principal barrier to absorption and led to significant systemic exposure to cidofovir. PMID- 9217247 TI - Effective treatment of experimental cytomegalovirus-induced encephalo-meningitis in immunocompromised rats with HPMPC. AB - Cytomegalovirus (CMV)-induced encephalomeningitis is a dramatic complication in patients with the acquired immunodeficiency syndrome (AIDS) and treatment of this infection remains a major clinical problem. In order to study the pathogenesis and treatment of CMV-induced encephalomeningitis, we experimentally induced intracranial rat CMV (RCMV) infection in rats that were immunosuppressed by total body X-irradiation. CMV infection was monitored by viral plaque assay for estimation of the viral load. CMV-induced pathology, the presence of CMV-infected cells, as well as the presence of T-lymphocytes and monocytes/macrophages were studied by histopathologic and immunohistochemical staining techniques. The meninges showed CMV infection in mononuclear infiltrative cells and in endothelium of small blood vessels 8 days after intracerebral inoculation. This was accompagnied by multiple haemorraghes and inflammatory cell infiltration. The infection and inflammatory response persisted for at least 21 days p.i. Animals were treated with (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine (HPMPC), 9 (1,3-dihydroxy-2-propoxymethyl)guanine (DHPG), hyperimmune serum (HIS) and both DHPG and HIS combined. Treatment with one dosage of HPMPC at 20 mg/kg effectively reduced virus titers. However, all other treatment modalities were not effective. In conclusion, the pathology of RCMV-induced encephalomeningitis in immunocompromised rats closely resembles that of AIDS patients. The infection is effectively treated by HPMPC. PMID- 9217249 TI - Evaluation of anti-AIDS drugs in conventional mice implanted with a permeable membrane device containing human T cells infected with HIV. AB - We now report the confirmation of the work of Hollingshead et al. (1995) on development of a cell based hollow fiber (HF) system for evaluating potential anti-AIDS drugs in vivo using conventional mice rather than SCID mice. CD4 +, CEM SS cells infected with HIV/1, strain RF, at a multiplicity of infection of 0.1 were placed into HFs. The fibers were implanted into the peritoneal cavity of outbred Swiss mice. Using this model, the antiviral activity of azidothymidine (AZT) at doses of approximately 150, 75 and 37.5 mg/kg/day was evaluated by administering AZT to the mice in drinking water. Upon fiber removal on day 6, AZT treatment was shown to significantly increase CEM cell viability over the untreated, virus control group and significantly reduced the levels of HIV p24 and HIV RT activity. PMID- 9217250 TI - Control of actin dynamics in cell motility. AB - Actin polymerization plays a major role in cell movement. The controls of actin sequestration/desequestration and of filament turnover are two important features of cell motility. Actin binding proteins use properties derived from the steady state monomer-polymer cycle of actin in the presence of ATP, to control the F actin/G-actin ratio and the turnover rate of actin filaments. Capping proteins and profilin regulate the size of the pools of F-actin and unassembled actin by affecting the steady-state concentration of ATP-G-actin. At steady state, the treadmilling cycle of actin filaments is fed by their disassembly from the pointed ends. It is regulated in two different ways by capping proteins and ADF, as follows. Capping proteins, in decreasing the number of growing barbed ends, increase their individual rate of growth and create a "funneled" treadmilling process. ADF/cofilin, in increasing the rate of pointed-end disassembly, increases the rate of filament turnover, hence the rate of barbed-end growth. In conclusion, capping proteins and ADF cooperate to increase the rate of actin assembly up to values that support the rates of actin-based motility processes. PMID- 9217251 TI - Voltage sensing in the PhoE and OmpF outer membrane porins of Escherichia coli: role of charged residues. AB - The porins PhoE and OmpF form anion and cation-selective pores, respectively, in the outer membrane of Escherichia coli. Each monomer of these trimeric proteins consists of a 16-stranded beta-barrel, which contains a constriction at half the height of the channel. The functional significance of a transverse electrical field that is formed by charged amino acid residues within the constriction zone was investigated. For this purpose, the PhoE residues R37, R75, K18 and E110 were substituted by neutral amino acids. The mutant pores allowed an increased permeation of beta-lactam antibiotics across the outer membrane in vivo, although the single channel conductance, measured in planar lipid bilayers, was not increased or even slightly decreased. Replacement of the positively charged residues resulted in a decreased voltage sensitivity, whereas the substitution of a negatively charged residue resulted in an increased voltage sensitivity. Similar substitutions in OmpF caused the opposite effects, i.e. the substitution of positive and negative charges resulted in increased and decreased voltage sensitivity, respectively. Together, the results suggest that opposite charges, i.e. positive charges in anion-selective and negative charges in cation-selective porins, act as sensors for voltage gating. PMID- 9217252 TI - Role of the carboxy-terminal phenylalanine in the biogenesis of outer membrane protein PhoE of Escherichia coli K-12. AB - Most bacterial outer membrane proteins contain a phenylalanine at their C terminus. It has been shown that this residue has an important role in the efficient and correct assembly of PhoE protein into the Escherichia coli outer membrane, since its substitution or deletion resulted in the accumulation of trypsin-sensitive monomers of this normally trimeric protein. Here, the role of the C-terminal Phe in the assembly of PhoE was studied in further detail. Immunocytochemical labelling on ultrathin cryosections revealed that a mutant PhoE protein that lacks the C-terminal Phe accumulates in the periplasm. However, when the expression levels of the altered species were reduced, the efficiency of outer membrane incorporation was increased and the lethal effects were alleviated. The role of the C-terminal Phe in protein folding, trimerization and outer membrane incorporation was further studied in vitro. Deletion of this residue interfered with the efficiency of the formation of an assembly-competent folded monomer, and the stability of this PhoE form was affected. The in vitro trimerization and insertion into outer membranes were not affected by the mutation. PMID- 9217253 TI - The equilibrium intermediate of beta-lactoglobulin with non-native alpha-helical structure. AB - It is generally considered that intermediates of protein folding contain partially formed native-like secondary structures. In contrast, we recently reported that the kinetic folding intermediate of bovine beta-lactoglobulin contains non-native alpha-helical structures. To understand the mechanism that stabilizes the non-native intermediate, we characterized by circular dichroism (CD) the equilibrium unfolding transition of beta-lactoglobulin induced by guanidine hydrochloride (Gdn-HCl) at pH 2 and 4 degrees C. The unfolding transition measured by near-UV CD preceded the transition measured by far-UV CD, indicating the accumulation of the intermediate state. The far-UV CD spectrum of the intermediate, obtained by global fitting analysis of the CD spectra in the presence of various concentrations of Gdn-HCl, was similar to the burst-phase intermediate observed in the refolding kinetics, and contained non-native alpha helical structures. Addition of 10% (v/v) 2,2,2-trifluoroethanol (TFE) increased the helical content of the equilibrium intermediate, although the protein still assumed the native structure in the absence of Gdn-HCl. A phase diagram of the conformational states, i.e. the alpha-helical intermediate, unfolded and native states, against the concentration of TFE and Gdn-HCl was constructed. This indicated that, because of the high helical preference of the amino acid sequence of beta-lactoglobulin, the helical region protrudes into the boundary between the native and unfolded states, resulting in non-monotonic accumulation of the helical intermediate upon equilibrium unfolding of the native beta-sheet structure. This is the first observation to indicate that a non-native alpha helical intermediate accumulates during equilibrium unfolding of a predominantly beta-sheet protein. PMID- 9217254 TI - Stability of type I collagen CNBr peptide trimers. AB - As indices of triple helix stability of type I collagen CNBr peptide homotrimers, deltaG degrees for monomer-trimer transitions and melting temperatures were obtained from circular dichroism measurements at increasing temperatures. The data were compared with the stability of the parent native molecule. Peptides were found to have a lower stability than the whole collagen molecule. The general implication is that the coordinated water molecules play a key role in determining collagen triple helical stability and high cooperativity at melting. Other factors (monomer stability, ionic and hydrophobic factors, variations of composition, specific sequences) could also contribute towards peptide stability; these factors could explain the data obtained in the case of peptide alpha1(I) CB3. PMID- 9217255 TI - Control of methylation spreading in synthetic DNA sequences by the murine DNA methyltransferase. AB - Methylation spreading, which involves a propensity for the mammalian DNA (cytosine-5)-methyltransferase to de novo methylate cytosine-guanine dinucleotides (CpGs) near pre-existing 5-methylcytosine bases, has been implicated in the control of numerous biological processes. We have assessed methylation spreading by the murine DNA methyltransferase in vitro using synthetic copolymers and oligonucleotides which differ only in their methylation state. Double-stranded oligonucleotides were found to undergo higher levels of de novo methylation overall than otherwise identical single-stranded oligonucleotides. This difference reflects the greater number of de novo methylatable cytosine bases in double-stranded than single-stranded sequences. All tested oligonucleotides containing pre-existing 5-methyl-cytosine(s) were de novo methylated at several fold the rates of non-methylated controls. No mammalian proteins besides the DNA methyltransferase were required for this observed enhancement of de novo methylation. Studies using oligonucleotides differing in patterns of pre-methylation showed that methylation spreading can be initiated by hemimethylated or duplex methylated CpGs indicating that recognition of 5-methylcytosine by the enzyme is sufficient to stimulate methylation spreading. Double and single-stranded oligonucleotides with several bases between CpGs underwent considerably more de novo methylation per CpG than sequences containing sequential uninterrupted methylatable sites. Spacing preferences by the DNA methyltransferase were also observed in hemimethylated oligonucleotides, suggesting that this is a general property of the enzyme. Although methylation spreading outside of CpG dinucleotides was relatively rare, single-stranded DNA incurred higher levels of de novo methylation at sites other than CpG as compared to double-stranded DNA. This indicates less specificity of methylation spreading in single-stranded sequences. Finally, enhanced de novo methylation in the presence of fully methylated CpG sites in double-stranded oligonucleotides was not as high as the rates of methylation of hemimethylated CpGs in otherwise identical oligonucleotides. These studies provide further elucidation of the mechanisms and regulation of the methylation spreading process and its potential role in the biological processes it influences. PMID- 9217256 TI - Partitioning of plasmid R1. The ParM protein exhibits ATPase activity and interacts with the centromere-like ParR-parC complex. AB - The parA system of plasmid R1 consists of two genes, parM and parR, and a cis acting centromere-like site parC. The ParM protein exhibits similarity with a superfamily of ATPases that includes actin, hsp70 and hexokinase. ParM was purified to near-homogeneity and assayed for in vitro ATPase activity. The wild type ParM protein was found to posses ATPase activity. Mutant ParM derivatives that exhibited decreased in vitro ATPase activity were non-functional in vivo, indicating that the ATP turnover by ParM is essential for correct plasmid partitioning. The mutant ParM proteins exhibited trans-dominance, suggesting that ParM participates as a structural component of the partitioning apparatus. The ATPase activity of ParM was activated slightly by the presence of ParR and activated to a much greater extent when ParR was bound to the centromere-like parC region. An analysis using the yeast two-hybrid system indicated that ParM and ParR interact, and demonstrated that ParR interacts with itself. Thus our results suggest a direct interaction of ParM and ParR at the natural partition site parC, and that the ATPase activity of ParM is specifically stimulated by this interaction. PMID- 9217257 TI - The quaternary geometry of transcription termination factor rho: assignment by chemical cross-linking. AB - Transcription termination factor rho from Escherichia coli is a ring-shaped homohexamer of 419 amino acid subunits and catalyzes an ATP-dependent release of nascent RNA transcripts. Previous chemical cross-linking studies suggested that the rho hexamer might have D3 symmetry with three isologous dimers as protomers. However, our recent mutational analysis of rho alongside its putative structural homology to F1-ATPase rather argued for C6 symmetry. To resolve this discrepancy, we have re-investigated the pattern of cross-linking of rho using various cross linkers with different functional groups and spacer lengths. Upon reaction with dimethyl suberimidate followed by SDS-polyacrylamide gel electrophoresis, rho protein generated a series of cross-linked oligomers up to hexamers, of which dimers migrated as distinct doublet bands of approximately equal intensities. However, the lower band became much stronger than the upper one with dimethyl adipimidate and difluorodinitrobenzene, and vice versa with disuccinimidyl glutarate, disuccinimidyl suberate and disulfosuccinimidyl tartarate. Furthermore, the trimeric products also produced doublet bands, whose relative intensities were again variable with cross-linkers, but in an inverse correlation with those of the dimer bands. These results combined with theoretical considerations support a C6 symmetry model in which cross-linking is assumed to occur stochastically at one of two alternative sites within each subunit interface with variable relative frequencies depending on cross-linkers. The D3 symmetry is excluded, for the putative trimeric subspecies should always retain mutually equal intensities in that case. Detailed inspections of the cross linking kinetics further revealed a moderate characteristic of C3 symmetry for the rho hexamer such that the collective as well as relative rates of cross linking at the two available sites could fluctuate between alternating interfaces. The final model designated as C3/6 is also compatible with other functional and structural properties known for rho. PMID- 9217258 TI - Structural characterisation of two forms of procyclic acidic repetitive protein expressed by procyclic forms of Trypanosoma brucei. AB - A procyclic acidic repetitive protein (PARP) fraction was purified from long-term cultures of Trypanosoma brucei procyclic forms by a solvent-extraction and reverse phase chromatography procedure. The PARP fraction yielded small quantities of a single N-linked oligosaccharide with the structure Man alpha1 6(Man alpha1-3)Man alpha1-6(Man alpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc (Man5GlcNAc2). Fractionation of PARP on Con A-Sepharose revealed that the majority (80 to 90%) of the PARP fraction did not bind to Con A and was composed of the parpA alpha gene product that contains repeats of -Glu-Pro-Pro-Thr- (GPEET PARP) and that lacks an N-glycosylation site. This form of PARP has not been previously identified at the protein-level. The minor Con-A-binding fraction was shown to be rich in the previously described form of PARP, encoded by the parpAbeta and/or parpB alpha genes, that contains a -Glu-Pro- repeat domain (EP PARP) and an N-glycosylation site. Analysis of longer and shorter-term cultures suggested that procyclic cells initially express predominantly EP-PARP that is gradually replaced by GPEET-PARP. Both forms of PARP were shown to contain indistinguishable glycosylphosphatidylinositol (GPI) membrane anchors, where the conserved GPI core structure is substituted by heterogeneous sialylated branched polylactosamine-like structures that are predicted to form a dense surface glycocalyx above which the polyanionic -Glu-Pro-Pro-Thr- and -Glu-Pro- repeat domains are displayed. The phosphatidylinositol (PI) component of the GPI anchor was shown to be a mixture of 2-O-acyl-myo-inositol-1-HPO4-(sn-1-stearoyl-2-lyso glycerol) and 2-O-acyl-myo-inositol-1-HPO4-(sn-1-octadecyl-2-lyso-glycerol), where the acyl chain substituting the inositol ring showed considerable heterogeneity. Mass spectrometric and light scattering experiments both suggested an average mass of approximately 15 kDa for GPEET-PARP, with individual glycoforms ranging from about 12 kDa to 20 kDa, that is consistent with its amino acid and carbohydrate composition. A measured translational diffusion coefficient of 3.9 x 10(7) cm2 s(-1) indicates that this molecule has a highly elongated shape. The possible functions of these unusual glycoproteins are discussed. PMID- 9217259 TI - Conformational changes due to calcium-induced calmodulin dissociation in brush border myosin I-decorated F-actin revealed by cryoelectron microscopy and image analysis. AB - Brush border myosin I (BBMI) is a single-headed molecular motor. Its catalytic domain exhibits extensive sequence homology to the catalytic domain of myosin II, while its tail lacks the coiled-coil nature of myosin II. The BBMI tail domain contains at least three IQ motifs and binds calmodulin. Addition of calcium removes one of these calmodulin light chains, with effects on ATPase activity and motility in in vitro assays. Using the techniques of cryoelectron microscopy and helical image analysis we have calculated three-dimensional (3D) maps of BBMI decorated actin filaments prepared in the presence and absence of calcium. The 3D maps describe a BBMI catalytic domain that is strikingly similar to the catalytic domain of myosin II subfragment 1 (S1), with the exception of a short amino terminal region of the heavy chain, which is absent from BBMI. The tail domains of BBMI and S1 are highly divergent in structure, continuing on from their respective motor domains with very different geometries. Addition of calcium to BBMI, and the concomitant loss of a calmodulin light chain, results in an extensive reorganization of mass in the tail domain. PMID- 9217260 TI - Human alpha-thrombin inhibition by the highly selective compounds N ethoxycarbonyl-D-Phe-Pro-alpha-azaLys p-nitrophenyl ester and N-carbobenzoxy-Pro alpha-azaLys p-nitrophenyl ester: a kinetic, thermodynamic and X-ray crystallographic study. AB - Kinetics, thermodynamics and structural aspects of human alpha-thrombin (thrombin) inhibition by newly synthesized low molecular weight derivatives of alpha-azalysine have been investigated. The thrombin catalyzed hydrolysis of N ethoxycarbonyl-D-Phe-Pro-alpha-azaLys p-nitrophenyl ester (Eoc-D-Phe-Pro-azaLys ONp) and N-carbobenzoxy-Pro-alpha-azaLys p-nitrophenyl ester (Cbz-Pro-azaLys-ONp) was investigated at pH 6.2 and 21.0 degrees C, and analyzed in parallel with that of N-alpha-(N,N-dimethylcarbamoyl)-alpha-azalysine p-nitrophenyl ester (Dmc azaLys-ONp). Decarboxylation following the enzymatic hydrolysis of these p nitrophenyl esters gave the corresponding 1-peptidyl-2(4-aminobutyl) hydrazines (peptidyl-Abh) showing properties of thrombin competitive inhibitors. Therefore, thermodynamics for the reversible binding of D-Phe-Pro-Abh, Cbz-Pro-Abh and Dmc Abh to thrombin was examined. These results are consistent with the minimum four step catalytic mechanism for product inhibition of serine proteinases. Eoc-D-Phe Pro-azaLys-ONp and Eoc-D-Phe-Pro-Abh display a sub-micromolar affinity for thrombin together with a high selectivity versus homologous plasmatic and pancreatic serine proteinases acting on cationic substrates. The three dimensional structures of the reversible non-covalent thrombin:Eoc-D-Phe-Pro-Abh and thrombin:Cbz-Pro-Abh complexes have been determined by X-ray crystallography at 2.0 A resolution (R-factor = 0.169 and 0.179, respectively), and analyzed in parallel with that of the thrombin:Dmc-azaLys acyl-enzyme adduct. Both Eoc-D-Phe Pro-Abh and Cbz-Pro-Abh competitive inhibitors are accommodated in the thrombin active center, spanning the region between the aryl binding site and the S1 primary specificity subsite. PMID- 9217261 TI - Crystal structure of a decameric complex of human serum amyloid P component with bound dAMP. AB - Serum amyloid P component (SAP) is a glycoprotein that binds in a calcium dependent fashion to a variety of ligands including other proteins, glycosaminoglycans and DNA. SAP is universally associated with the amyloid deposits in all forms of amyloidoses including Alzheimer's disease. Small molecule ligands that displace SAP from amyloid fibrils and thereby expose the fibrils to proteolytic clearance mechanisms hold potential as drugs for the prevention and treatment of amyloidosis. We have carried out a screen for novel SAP ligands and have identified 2'-deoxyadenosine-5'-monophosphate (dAMP) as a ligand. The crystal structure of the SAP-dAMP complex determined at 2.8 A resolution (R = 0.232, R(free) = 0.252) reveals a decamer in which all interactions between SAP pentamers are mediated by the ligand. The stability of the decamer in solution has been demonstrated by gel filtration chromatography. The two calcium ions of SAP are bridged by the dAMP phosphate group and five hydrogen bonds are formed between the protein and the ligand, including specific interactions made by the adenine base. This mode of dAMP binding is not compatible with the nucleotide being part of double-helical DNA. The SAP-dAMP decamer is stabilized mainly by base-stacking of adjacent ligand molecules and possibly by electrostatic interactions involving the dAMP phosphate groups; decamerization buries 1000 A2 (2.6%) of the pentamer solvent-accessible surface. Ligand-induced decamerization of SAP, which utilizes the high cooperativity of a multiple-site interaction, may be a strategy to overcome the problems for drug design associated with the rather modest affinities of SAP for small-molecule ligands. PMID- 9217262 TI - The solution structure of the pleckstrin homology domain of mouse Son-of sevenless 1 (mSos1). AB - The solution structure of the pleckstrin homology (PH) domain of mouse Son-of sevenless 1 (mSos1), a guanine nucleotide exchange factor for Ras, was determined by multidimensional NMR spectroscopy. The structure of the mSos1 PH domain involves the fundamental PH fold, consisting of seven beta-strands and one alpha helix at the C terminus, as determined for the PH domains of other proteins. By contrast, the mSos1 PH domain showed two major characteristic features. First, the N-terminal region, whose amino acid sequence is highly conserved among Sos proteins, was found to form an alpha-helix, which interacts with the beta-sheet structure of the fundamental PH fold. Second, there is a long unstructured loop between beta3 and beta4. Furthermore, the mSos1 PH domain was found to bind phosphatidylinositol-4,5-bisphosphate by a centrifugation assay. The addition of inositol-1,4,5-trisphosphate to the mSos1 PH domain induced backbone amide chemical shift changes mainly in the beta1/beta2 loop and the N- and C-terminal parts of the long beta3/beta4 loop. This inositol-1,4,5-trisphosphate-binding mode of the mSos1 PH domain is somewhat similar to those of the PH domains of pleckstrin and phospholipase Cdelta1, and is clearly different from those of other PH domains. PMID- 9217263 TI - Automated analysis of protein NMR assignments using methods from artificial intelligence. AB - An expert system for determining resonance assignments from NMR spectra of proteins is described. Given the amino acid sequence, a two-dimensional 15N-1H heteronuclear correlation spectrum and seven to eight three-dimensional triple resonance NMR spectra for seven proteins, AUTOASSIGN obtained an average of 98% of sequence-specific spin-system assignments with an error rate of less than 0.5%. Execution times on a Sparc 10 workstation varied from 16 seconds for smaller proteins with simple spectra to one to nine minutes for medium size proteins exhibiting numerous extra spin systems attributed to conformational isomerization. AUTOASSIGN combines symbolic constraint satisfaction methods with a domain-specific knowledge base to exploit the logical structure of the sequential assignment problem, the specific features of the various NMR experiments, and the expected chemical shift frequencies of different amino acids. The current implementation specializes in the analysis of data derived from the most sensitive of the currently available triple-resonance experiments. Potential extensions of the system for analysis of additional types of protein NMR data are also discussed. PMID- 9217264 TI - The rate of isomerisation of peptidyl-proline bonds as a probe for interactions in the physiological denatured state of chymotrypsin inhibitor 2. AB - There are four peptidyl-proline bonds in the 64-residue protein chymotrypsin inhibitor 2 (CI2), all of which are in the trans conformation in the native structure. The isomerisation of one or more of these peptidyl-proline bonds to the cis conformation in the denatured state gives rise to heterogeneity, leading to both fast and slow-folding species. The refolding of the fast-folding species, which has all trans peptidyl-proline bonds, is much faster than that of the slow folding species, which have one or more cis peptidyl-proline bonds. In CI2, the slow-folding species can be classified into two groups by their rates of refolding, temperature-dependence, pH-dependence and [GdmCl]-dependence of the rate constants and the effect of peptidyl-prolyl isomerase on the rate constants. The replacement of Pro6 by Ala removes one of the slow refolding phases, suggesting that the cis peptidyl-Pro6 conformation is solely responsible for one of the slow-folding species. Pro6 is located in a region of the protein where non random interactions have been found in a series of N-terminal fragments of CI2 (residues 1 to 13, 1 to 25, 1 to 28 and 1 to 40). In addition, NMR studies on a mutant fragment, (1-40)T3A, have confirmed that this non-native interaction is associated with the bulky side-chain of Trp5. The atypical rate of cis to trans isomerisation of the peptidyl-Pro bond is indicative of the presence of a similar hydrophobic cluster in the physiological denatured state of intact CI2. PMID- 9217266 TI - Protein thermal stability, hydrogen bonds, and ion pairs. AB - Researchers in both academia and industry have expressed strong interest in comprehending the mechanisms responsible for enhancing the thermostability of proteins. Many and different structural principles have been postulated for the increased stability. Here, 16 families of proteins with different thermal stability were theoretically examined by comparing their respective fractional polar atom surface areas and the number and type of hydrogen bonds and salt links between explicit protein atoms. In over 80% of the families, correlations were found between the thermostability of the familial members and an increase in the number of hydrogen bonds as well as an increase in the fractional polar surface which results in added hydrogen bonding density to water. Thus increased hydrogen bonding may provide the most general explanation for thermal stability in proteins. The number of ion pairs was also found to increase with thermal stability in two-thirds of the families tested; however, their rate of addition was only about one-sixth that for internal hydrogen bonds amongst the protein atoms. The preferred residue exchanges and surface atom types useful in engineering enhanced stability were also examined. PMID- 9217265 TI - Directed mutagenesis shows that the preceding region of the chloroplast fructose 1,6-bisphosphatase regulatory sequence is the thioredoxin docking site. AB - The alignment of the six higher plant photosynthetic fructose-1,6-bisphosphatases (FBPases) so far sequenced shows a lack of homology in the region which just precedes the cluster engaged in light modulation. Earlier experiments suggested that this region is the docking point in FBPase-thioredoxin (Trx) binding, and could be responsible for the interspecific differences in the enzyme-Trx interaction and Trx ability for FBPase activation. Using a pea chloroplast FBPase coding cDNA, we have prepared two chimeric clones for FBPase. One of them (pDELFBP) shows a deletion of the 17 amino acids (Leu154 to Glu170) coding sequence, whereas in the second (pPFBPW) the above sequence was substituted by the corresponding one of the wheat enzyme. After Escherichia coli overexpression in pET-3d and later purification, both modified FBPases showed FBPase activity when determined under non-reducing conditions. However, only DELFBP lost the Trx f modulatory effect, indicating the important role played by this fragment in FBPase-Trx interaction and activity. Under these conditions the substituted PFBPW enzyme retains FBPase activity, even though clearly diminished. Superose 12 filtration experiments after preincubating the wild-type and modified FBPases with Trx f, showed the existence of an enzyme-Trx f binding with the wild-type and the substituted PFBPW, but not with the deleted DELFBP protein. Similarly, gradient PAGE under native conditions, followed by Western blot and developing with FBPase and Trx f antibodies, indicated the existence of such a binding between the wild-type and PFBPW, on the one hand, and both Trxs f and m, on the other, although never with the deleted DELFBP enzyme. These results show the central role played by the regulatory site preceding fragment of chloroplast FBPase in its binding with Trx. Computer-aided tridimensional models for the wild type and modified FBPases are proposed. PMID- 9217267 TI - Neurotransmitter aberrations in schizophrenia: new perspectives and therapeutic implications. AB - The dopamine hypothesis has dominated schizophrenia research for decades but is now yielding to a more diversified view, where the interaction of several neurotransmitters in complex circuitries is under scrutiny. Especially, glutamatergic and serotonergic mechanisms are attracting attention. However, the role of dopamine also needs further exploration and may still turn out to have novel therapeutic applications. In the present minireview an attempt is made to integrate preclinical and clinical data on neurotransmitter aberrations in schizophrenia and to discuss their therapeutic implications. PMID- 9217268 TI - Biochemical synthesis, purification and preliminary pharmacological evaluation of normorphine-3-glucuronide. AB - Normorphine was synthesised from morphine by thermal decomposition of an N-alpha chloroethylchloroformate adduct, and purified (> 98% purity) using semi preparative HPLC with ultraviolet detection. Normorphine-3-glucuronide (NM3G) was biochemically synthesised using the substrate normorphine, uridine diphosphoglucuronic acid and Sprague-Dawley rat liver microsomes in a 75% yield (relative to normorphine base). The synthesised NM3G was purified by precipitation and washing with acetonitrile. Determinations of purity using HPLC with electrochemical and ultraviolet detection confirmed that the NM3G produced was of high (> 99%) purity. Mass spectrometry, fourier transform infrared spectrophotometry and nuclear magnetic resonance spectrometry confirmed the structure, especially placement of the glucuronide moiety at the 3-phenolic position and not at the 17-nitrogen. Administration of NM3G by the intracerebroventricular (icv) route to rats in doses of 2.5 and 7.5 microg resulted in the development of central nervous system (CNS) excitatory behavioural effects including myoclonus, chewing, wet-dog shakes, ataxia and explosive motor behaviour. At an icv dose of 7.5 microg, NM3G also induced short periods of tonic-clonic convulsive activity. Thus, NM3G elicits CNS excitation following supraspinal administration in a manner analogous to morphine-3 glucuronide (M3G), the major metabolite of morphine (1). Further studies are required to determine whether NM3G attenuates morphine-induced antinociception in a similar manner to M3G. PMID- 9217269 TI - Effects of biochanin A on the growth and differentiation of myeloid leukemia WEHI 3B (JCS) cells. AB - The effects of biochanin A on the growth and differentiation of a recently characterized myeloid leukemia cell line WEHI-3B (JCS) were investigated. Biochanin A not only inhibited the growth of JCS cells in a dose-dependent manner (0 - 200 microM) but also induced the morphological differentiation of JCS cells. The phagocytic activity of biochanin A-treated JCS cells was also increased. Flow cytometric analysis showed that the expression of macrophage differentiation markers Mac-1 and F4/80 was up-regulated in biochanin A-treated JCS cells. The expression level of Mac-1 was higher than that of F4/80. The expression of cytokine genes was studied by reverse transcription-polymerase chain reaction (RT PCR) and cycle titration. mRNA levels of IL-1alpha, IL-1beta and IL-4 were found to be up-regulated at 46 hours after incubation of JCS cells with biochanin A. Although the expression of LIF was also up-regulated, the LIF receptor gene was not expressed in the uninduced or induced JCS cells. Our results suggest that IL 1alpha, IL-1beta and IL-4 may act on the later stage of biochanin A-mediated differentiation of JCS cells. PMID- 9217270 TI - Handling stress does not affect the expression of hepatic heat shock protein 70 and conjugation enzymes in rainbow trout treated with beta-naphthoflavone. AB - A response in heat shock protein 70 (hsp 70) expression in the beta naphthoflavone (BNF) treated rainbow trout (Oncorhynchus mykiss) corresponded to altered metabolic status of the liver as evidenced by the lower phosphoenolpyruvate carboxykinase (PEPCK), lactate dehydrogenase and 3 hydroxyacylcoA dehydrogenase activities. The BNF-induced increase in hsp70 levels and conjugation enzyme activities (phase I and phase II) were not modified by handling stress. Indeed handling stress did not affect either hsp 70 levels or conjugation enzyme activities in trout liver. The decrease in hepatic PEPCK activity in the BNF group may be responsible for the attenuation of the increase in liver glucose concentration after a 3 min handling stress in this species, suggesting that BNF affects liver gluconeogenic capacity in this species. Handling stress elicited a plasma cortisol and glucose response in both the sham and BNF group, however, the cortisol response with BNF was erratic compared with the sham, implying alterations in the cortisol dynamics post-stress. These results show for the first time that BNF affects cellular metabolic responses to stress and suggests the possibility of using hsp 70 as a biomarker for toxic effects in trout. PMID- 9217272 TI - Suppression of polarity, locomotion and F-actin levels of Walker carcinosarcoma cells by the inhibitor CI-959. AB - Locomotor activity of tumor cells is an important factor for the capacity for invasion and metastasis. Therefore, inhibitors interfering with cellular mechanisms regulating spontaneous cell locomotion are of particular interest for cancer therapy. CI-959, a new benzothiophene cell activation inhibitor, has the capacity to suppress spontaneous polarity and locomotion of Walker carcinosarcoma cells. Suppression of polarity and locomotion was closely associated with a reduction in the relative amount of F-actin. The mechanisms involved in suppression of motility are Ca2(+)-independent and not related to cell-substratum adhesion. Walker carcinosarcoma cells appear to be able to locomote at very low (nM) levels of free [Ca2+]i. PMID- 9217271 TI - Extraordinary potency of a novel delta opioid receptor agonist is due in part to increased efficacy. AB - A new cyclic opioid peptide of sequence Tyr-D-Pen-Gly-Phe-Cys-Phe (HBP2) was examined in the mouse isolated vas deferens (MVD) bioassay. Studies with receptor selective opioid antagonists showed the peptide to be highly selective for delta opioid receptors. HBP2 and the standard delta agonist DPDPE were simultaneously compared using the technique of partial irreversible receptor blockade; data were analyzed using the operational model of pharmacologic agonism. HBP2 was approximately 160 times as potent as DPDPE; estimation of the affinity and efficacy of the two peptides revealed that the potency increase was due to a 5.3 fold increase in efficacy, as well as a 37-fold increase affinity. This contrasts with our previous findings with other cyclic enkephalin analogs, in which increased affinity was achieved without a change in apparent efficacy. Analysis of concentration-response curve shape suggested in addition the possibility of heterogeneity in transduction mechanisms for MVD delta receptors. PMID- 9217273 TI - The effect of 1,25(OH)2 vitamin D3 on CD4+/CD8+ subsets of T lymphocytes in postmenopausal women. AB - The effect of exogenous 1,25(OH)2 vitamin D3 (1,25(OH)2D3) on the CD3+, CD4+ and CD8+ subsets (counts/ul) of T lymphocytes was investigated in two randomized groups of postmenopausal women. Group one (16 subjects) received 1 ug/day of the secosteroid for 14 days, while group two (14 participants) was treated with 2 ug/day for the same period. The placebo group comprised another 10 postmenopausal women. Compliance of the treatment was controlled by serum intact parathyroid hormone (PTH) levels, which markedly declined at the end of the treatment (p<0.01 for both doses). The vitamin D status of the women before the treatment was defined by serum 25(OH) vitamin D (25(OH)D) levels. The lower dose of the secosteroid did not change any of the measured immune parameters. After a higher dose of 1,25(OH)2D3 the mean values of CD3+ and CD8+ increased (p<0.05 for the both parameters), but no changes in total lymphocytes and the CD4+ subset were observed. There were no correlations between the immune response (delta CD3+, delta CD4+ and delta CD8+) and basal circulating 25(OH)D. Briefly, then, 1,25(OH)2D3 slightly but significantly increases CD3+ and CD8+ subsets independently on the initial vitamin D status of the postmenopausal women. PMID- 9217274 TI - Effect of the P-glycoprotein inhibitor, SDZ PSC 833, on the blood and brain pharmacokinetics of colchicine. AB - The effect of the multidrug resistance-reversing agent, SDZ PSC 833, on blood and brain pharmacokinetics of a P-glycoprotein substrate, colchicine, was investigated using simultaneous blood and brain microdialysis in freely moving rats. The use of microdialysis for pharmacokinetic studies was validated by comparing the blood concentrations of colchicine obtained by microdialysis with those obtained by direct blood sampling. The rats received either SDZ PSC 833 (2.3 mg/kg i.v. bolus followed by 16.7 microg/min/kg i.v. infusion during all the experiment) and colchicine (1 mg/kg i.v. bolus followed by 12.5 microg/min/kg i.v. infusion during 2 hours) or colchicine alone (the same dosage with SDZ PSC 833 vehicle). The SDZ PSC 833 treatment resulted in important modifications of colchicine blood pharmacokinetics: the unbound colchicine blood concentration at steady-state was enhanced from 149.6 +/- 9.9 to 333.5 +/- 81.7 ng/ml indicating a two-fold decrease in colchicine clearance. Moreover the coadministration of SDZ PSC 833 increased the brain penetration of colchicine by a factor of 10, at least. This enhancement could not be exactly assessed because the brain dialysate concentrations of control group were below the limit of detection. Nevertheless, the large increase of colchicine brain penetration is consistent with the hypothesis that SDZ PSC 833 is able to inhibit the P-glycoprotein pump present at the blood-brain barrier. PMID- 9217275 TI - Estrogen and parathyroid hormone regulate insulin-like growth factor binding protein-4 in SaOS-2 cells. AB - The effect of 17beta-estradiol and parathyroid hormone (PTH) on the expression of insulin-like growth factor-binding protein-4 (IGFBP-4) messenger RNA (mRNA) was studied in the cultured human osteoblast-like SaOS-2 cells. Treatment of SaOS-2 cells with PTH for 3 h caused 3.3-fold increase in IGFBP-4 mRNA levels which was determined by reverse transcription-polymerase chain reaction. 17beta-Estradiol had no effect on either the stimulation of mRNA level by PTH or the basal level. Together with our previous report that 17beta-estradiol inhibits the PTH-induced reduction of IGFBP-4 proteolysis in these cells, the results obtained may help to explain the mechanisms of determining IGFBP-4 availability by systemic hormones in osteoblast cells. PMID- 9217276 TI - Expression of cytochrome P450 isoforms in rat hepatic stellate cells. AB - The current study evaluated the expression and the inducibility of cytochrome P450 isoforms in rat hepatic stellate cells (HSCs). Immunoblotting study revealed that HSCs expressed several P450s and CYP2C11, 3A2, and 2D1 were major isoforms. The levels of CYP2B1, 2C11, 2D1, 2E1, and 3A2 in HSCs were 14 - 38% of those in hepatocytes. CYP1A2 content was similar in each cell type. These P450 levels in HSCs gradually decreased during culture as seen in hepatocytes; the level of CYP3A2 rapidly, whereas that of CYP2D1 slowly decreased. Phenobarbital, a typical inducer of CYP3A2 and 2B1 increased CYP3A2 level as well, but had less potency in the induction of CYP2B1 in HSCs. These results indicate that multiple P450 isoforms were present in HSCs, but their content and inducibility were different between HSCs and hepatocytes. PMID- 9217277 TI - Different experimental conditions which regulate type II 5'-deiodinase mRNA in rat Harderian gland. AB - In the present study, we describe the modifications in the expression of type II 5'deiodinase activity (5'D) in Xenopus laevis oocytes by injection of polyadenylated (poly A) mRNA from hypothyroid rat Harderian gland. The time course study showed that the expression of the enzyme was dependent on time. Thus, enzyme activity was observed in oocytes 6 and 12 hours after the injection with poly A mRNA, reaching a maximal value at 24 hours. The activity was partially inhibited by 6-n-propyl-thiouracil, completely inhibited by iopanoic acid and exhibited a higher affinity for the T4 (Km=1.5 nM) than rT3 (Km=20 nM). The expression of the enzyme was modified in different experimental conditions: (a) exhibited diurnal variations with maximal peak values at night, (b) was inhibited by light at night and, (c) was activated by isoproterenol. On the other hand, we have also identified, for the first time, the size of mRNA capable of inducing 5'D in rats. PMID- 9217278 TI - Effect of triiodothyronine on glucose transport in rat adipocytes. AB - The in vitro effect of thyroid hormones on glucose transport in insulin stimulated muscle cells or adipocytes is still unclear. The objective of the present study was to assess the direct effect of 3,3',5-triiodothyronine (T3) on glucose transport and on the translocation of insulin-regulatable glucose transporter (GLUT4) in insulin-stimulated rat adipocytes. This evaluation was performed using an in vitro assay to avoid the well-known systemic effects of this hormone ( e.g.: hyperinsulinemia). Adipocytes were isolated from epididymal adipose tissue of Sprague-Dawley rats. Glucose transport assay and immunoblot analysis of GLUT4 were carried out in insulin-stimulated and unstimulated adipocytes after treating with or without T3. The results were as follows; 1) T3 inhibited the glucose transport in insulin-stimulated and unstimulated adipocytes in a dose-dependent manner. 2) T3 decreased the maximal response level (Vmax) but did not alter the sensitivity (Km) of glucose transport to insulin. 3) T3 did not affect the translocation of GLUT4 from the intracellular pool to the plasma membrane. We concluded that T3 inhibits the glucose transport in insulin stimulated adipocytes in a post-receptor level without affecting the translocation of GLUT4 from the intracellular pool to the plasma membrane. This suggests that T3 acts by decreasing the intrinsic activity or the accessibility of GLUT4 in the plasma membrane. PMID- 9217279 TI - Comparison of susceptibility to apoptosis induced by rhTNF-alpha and cycloheximide between human circulating and exudated neutrophils. AB - To determine whether exudated neutrophils differ from circulating ones in their apoptosis, rhTNF-alpha plus cycloheximide-induced apoptosis in human salivary neutrophils was compared to that in human neutrophils in peripheral blood. Concomitant treatment of peripheral blood neutrophils with rhTNF-alpha and cycloheximide-induced apoptosis in blood neutrophils within 3 hr, as evaluated both by light microscopic changes characteristic to apoptosis and by DNA fragmentation, whereas the same treatment failed to induce any apoptosis in salivary neutrophils. These results indicate that the exudation of neutrophils from blood into tissue is associated with marked changes in their functions such as alteration in their sensitivity to apoptosis-inducing stimuli. PMID- 9217281 TI - Induction of ICE and inhibition of c-fos, jun D and zif 268 in 12-month old spontaneously hypertensive rats. AB - A semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) assay was used to examine ICE, c-fos, jun D and zif 268 mRNA expression in the aortic and renal artery of 12-month old SHRs and wistar rats. Using this assay system, it was observed that the levels of aortic and renal artery expression of ICE were markedly higher in SHRs than in wistar rats. In contrast, the aortic and renal artery expression of immediate early genes (IEGs), c-fos, jun D and zif 268, were significant lower in SHRs than in wistar rats. Thus, our results suggest that differential regulation of death gene ICE and IEGs such as c-fos, jun D and zif 268 might be involved in the mechanism of pathogenesis of hypertension. PMID- 9217280 TI - Functional characterization of peripheral muscarinic subtypes in anesthetized cats. AB - This investigation was undertaken to characterize the muscarinic receptor subtypes involved in methacholine-induced vasodilation, vagal bradycardia, neurally-evoked sudomotor responses and sympathetic muscarinic ganglionic transmission in anesthetized cats. Dose-response curves were constructed using the putatively selective antagonists pirenzepine (M1), gallamine (M2) and 4-DAMP (M3: 4-diphenyl-acetoxy-N-methylpiperidine) and compared with the non-selective blocker, atropine. Methacholine hypotension and evoked sudomotor responses exhibited an M3 muscarinic receptor profile with the following potency relationships: atropine > or = 4-DAMP > pirenzepine >> gallamine. Vagal bradycardia (M2) was antagonized by gallamine and exhibited a lower relative sensitivity to 4-DAMP when corrected for atropine effect. Pirenzepine was inactive in inhibition of bradycardia but was highly potent against transmission in the sympathetic ganglion (M1) with the following potency relationships: atropine > or = pirenzepine > 4-DAMP >> gallamine. In comparison with atropine, 4 DAMP exhibited a significantly lower potency for M1 and M2 muscarinic receptors as compared to its effect on the M3 muscarinic receptor subtypes. PMID- 9217282 TI - Inhibition of nitric oxide does not affect reperfusion-induced myocardial injury, but it prevents lipid peroxidation in the isolated rat heart. AB - To examine if inhibition of nitric oxide (NO) synthesis influences myocardial ischemia-reperfusion injury, male Sprague Dawley rats were administered the NO synthesis inhibitor N -nitro-L-arginine methyl ester (L-NAME, 10 mg/kg, i.p.) or saline 6 hours prior to excising the heart and aorta. Aortic ring contractile response to norepinephrine (NE) was more pronounced and relaxation in response to acetylcholine was abolished in L-NAME-treated group (P<0.05 vs. saline-treated group), indicating inhibition of NO synthesis in the vascular tissues. In the isolated perfused Langendorff hearts, force of cardiac contraction (FCC) and coronary perfusion pressure (CPP) were higher and coronary flow was lower in the L-NAME-treated group, again suggesting inhibition of NO synthesis. Global ischemia (40 min) followed by reperfusion (30 min) resulted in a decrease in FCC and coronary flow and an increase in CPP in all hearts. Myocardial CK also decreased similarly in all hearts. However, ischemia-reperfusion-induced decline in myocardial superoxide dismutase (SOD) activity and increase in malondialdehyde were prevented in the L-NAME-treated group (P<0.01 vs. saline-treated hearts). Thus treatment with L-NAME with resultant inhibition of NO synthesis does not affect ischemia-reperfusion-induced cardiac dysfunction and injury in the isolated rat hearts, although the reduction in SOD activity and the rise in lipid peroxidation following reperfusion are attenuated. PMID- 9217283 TI - KC8851, a tedisamil analogue with mixed channel blockade, exhibits antiarrhythmic properties against ischemia- and electrically-induced arrhythmias. AB - KC8851, a structural analogue of tedisamil, has previously been found to exhibit mixed blockade of K+ and Na+ currents in isolated rat ventricular myocytes. We have now investigated the antiarrhythmic actions of this compound in the anaesthetized rat and isolated rat heart. In the rat, KC8851, at concentrations as low as 0.2 micromol kg(-1) min(-1), widened the QT intervals of the ECG and prolonged the effective refractory period in a dose-dependent manner. Such actions were consistent with blockade of repolarizing K+ currents. At relatively higher doses (above 0.5 micromol kg(-1) min(-1)), KC8851 increased RSh amplitude suggesting blockade of Na+ currents. The compound was found to be effective against occlusion-induced arrhythmias at doses of 0.5 to 2 micromol kg(-1) min( 1). In isolated hearts, the effects of KC8851 on PR and QRS intervals were potentiated by elevated concentrations of K+ and H+. Overall, KC8851 was found to exhibit antiarrhythmic actions consistent with inhibition of both K+ and Na+ currents. PMID- 9217284 TI - Specific detection of kappa light chain in uric acid stones. AB - Proteins were extracted from uric acid stones with 6M guanidine chloride (pH 8.5), which were successively developed by 12% polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE). Amino acid sequence analysis of each band on SDS-PAGE revealed that major components in uric acid stones were immunoglobulin alpha heavy and kappa light chains. Although immunoglobulin heavy chain (gamma and mu, as well as alpha) and a kappa light chain were clearly detected in uric acid stones by Western blotting using their specific antibodies, no lambda chains whatsoever could be detected. The results suggest that immunoglobulins selectively containing kappa light chain might have specific functions in uric acid stone formation as stone matrices. PMID- 9217285 TI - Absence of ET(B)-mediated contraction in Piebald-lethal mice. AB - Activation of the endothelin (ET) ET(B) receptor can mediate opposite effects, endothelium-dependent vasodilation but also direct vasoconstriction. So far one gene encoding an ET(B) receptor has been identified and associated with endothelium-dependent relaxation. It has been suspected that the presence of another ET(B) gene could explain ET(B)-mediated contraction. The goal of the present study was to evaluate in Piebald-lethal (s[1]) mice, a naturally occurring mutant with deletion of the known ET(B) receptor gene, whether ET(B) receptor-mediated constriction is lost. Piebald-lethal (s[1]) mice, in contrast to control mice, completely lacked ET(B) specific ligand binding. The pressor effect of the ET(B) receptor selective agonist sarafotoxin S6c was completely absent. In vitro, contraction of stomach strips induced by sarafotoxin S6c was also abolished in Piebald-lethal (s[1]) mice. These results demonstrate the responsibility of the known ET(B) receptor gene in ET(B)-mediated constriction. PMID- 9217286 TI - Metabolic defects caused by treatment with the tetrahydropyridine analog of haloperidol (HPTP), in baboons. AB - Mounting evidence suggests that compromised cellular energy production is a major contributor to idiopathic and drug-induced degenerative processes. Our interest in neurotoxins have prompted us to examine in the baboon the effects of HPTP, the tetrahydropyridine dehydration product of haloperidol, on urinary chemical markers that reflect defects in mitochondrial respiration. Urinary dicarboxylic acid and conjugate profiles, similar to those seen in humans with inborn errors of mitochondrial metabolism and toxin-induced Jamaican vomiting sickness (JVS) were observed in the treated baboons. We interpret these results as evidence that HPTP and/or HPTP metabolites inhibit mitochondrial respiration in the baboon and speculate that analogous effects may occur in haloperidol-treated individuals. PMID- 9217287 TI - Effects of neurotropic pyrimidine heterocyclic compound, MS-430, on cultured hepatic parenchymal and stellate cells. AB - MS-430 is a novel synthetic pyrimidine derivative that stimulates regeneration of the nerve as a promoter for various growth factors such as epidermal growth factor (EGF) and nerve growth factor, and differentiation of astrocytes. The effects of MS-430 on the liver were tested using hepatocytes and stellate cells in primary culture isolated from rats. MS-430 enhanced EGF-induced DNA synthesis in hepatocytes while it alone failed to increase the basal DNA synthesis. Albumin mRNA expression in the cells and its amount in the medium were not changed by addition of EGF or MS-430 alone or both. Basic fibroblast growth factor (bFGF) increased DNA and but not collagen synthesis by hepatic stellate cells. Addition of MS-430 inhibited DNA synthesis by hepatic stellate cells at either presence or absence of bFGF, and collagen synthesis at the presence of bFGF. However, MS-430 had no effects on basal or bFGF-stimulated TGFbeta mRNA expression in the cells. These results suggest that MS-430 stimulated proliferation of hepatocytes as a comitogen for EGF without affecting albumin synthesis, and suppressed proliferation of activated hepatic stellate cells and their collagen synthesis without affecting TGFbeta expression. PMID- 9217288 TI - Immunotherapy for insulin-dependent diabetes mellitus in the "BB" rat. AB - We have previously reported that weekly administration of the adenosine deaminase inhibitor, 2'-deoxycoformycin (dCF), reduces the incidence of insulin-dependent diabetes mellitus (IDDM) in the BB Wistar rat, and this effect is likely due to immunosuppression by dCF. In the present study, we examined the effect of altering the dose and scheduling of dCF on prevention of IDDM in the BB rat. When rats were treated from day 25 of age with 2.5, 4, or 10 mg of dCF/kg/week, the percentage of diabetes-free animals at 120 days of age was 40, 60, and 80% respectively, compared with 10% for control animals, demonstrating increased protection against IDDM with increased dCF dose. Histological assessment of the pancreata from animals that became diabetic revealed a marked mononuclear infiltrate and a loss of positive staining for beta cell granules. In contrast, pancreata from animals that remained diabetes-free appeared normal. Protection against IDDM by dCF was time dependent and only occurred if treatments were initiated by day 30 of age. In addition, the protective effect persisted after drug withdrawal. Further studies are required to determine the optimum duration of therapy with dCF to prevent IDDM and to examine the immunological mechanism responsible for this effect. PMID- 9217289 TI - Primary cultured chondrocytes of different origins respond differently to bFGF and TGF-beta. AB - Responses of rib and ear chondrocytes to basic fibroblast growth factor (bFGF) and transforming growth factor-beta (TGF-beta) were investigated using high density primary culture isolated from the same rabbit. Degrees of tritiated thymidine, leucine, and proline incorporation were used as indicators of DNA, protein and collagen syntheses, respectively. 10 ng/ml bFGF increased thymidine, proline, and leucine incorporation into rib, but not ear, chondrocytes. 1 ng/ml TGF-beta enhanced thymidine incorporation into both chondrocytes but did not affect proline or leucine incorporation into the ear cells. When both growth factors were added simultaneously, both cells showed rises in syntheses of DNA, protein and collagen. Incorporation of [35S]sulfate, used as indicator of proteoglycan synthesis, was elevated by TGF-beta but was reduced by bFGF especially in the rib cells. This inhibitory effect of bFGF was not reversed by cotreatment with TGF-beta in both cell types. Thus, the origin and cellular differentiation states of chondrocytes seem to cause different responses to these growth factors. PMID- 9217290 TI - Glucocorticoid receptors in human peripheral blood mononuclear cells in relation to age and to sport activity. AB - Glucocorticoid receptors (GR) are ubiquitous molecules and are present also in the hippocampus and in several other nervous and immune tissues. Peripheral blood mononuclear cells (PBMCs) are a good model for studies of GR in humans. Glucocorticoids are important for maintaining cellular and humoral homeostasis and are key mediators of neuroendocrine-immune regulatory interactions. The increase of cortisol is immunosuppressive and reduces GR concentration both in nervous and immune systems. Variation of glucocorticoids in healthy aged subjects and athletes has been shown. Prompted by these results, we have investigated in man a possible relationship between GR binding capacity in the PBMCs and age, in relation also to plasma testosterone and cortisol. The same parameters have been examined in a group of soccer players for comparison with the sedentary group. GR binding capacity was higher in younger subjects than in older ones, and lower in the group of athletes than in the younger and older sedentary subjects. In the sedentary group a negative correlation was present between GR binding capacity and age. Plasma cortisol was higher and testosterone lower in the athletes; they were negatively correlated in athletes and positively correlated in the sedentary subjects. The results for athletes agree with their lower anabolic/catabolic balance. The mechanism of reduced GR levels in relation to age and sport activity could involve a loss or an involution of receptor synthesis. However other possibilities, such as altered distribution of lymphocyte subpopulations with different receptor concentrations and with different cytokine production, cannot be excluded. Several neuroendocrine-immune interactions could be responsible for reduced GR levels with age and sport activity in man. PMID- 9217291 TI - 5-HT receptor subtypes involved in the serotonin-induced inhibition of L-leucine absorption in rabbit jejunum. AB - The aim of the present study was to determine the 5-HT receptor subtypes involved in the serotonin-induced inhibition of L-leucine absorption across rabbit jejunum in vitro. A number of agonists and antagonists were used to characterize the receptors through which serotonin inhibits this absorption. The results show that 2.5x10(-6) M 5-HT inhibits the amino acid absorption by about 20%. The 5-HT receptor agonists, alpha-methyl-5-HT (5-HT2), 2-methyl-5-HT (5-HT3) and zacopride (5-HT4) at concentrations 2.5x10(-6) and 2.5x10(-5) M produced 10-30% inhibition on L-leucine intestinal absorption. 5-carboxyamidotryptamine (5-HT1) did not produce any inhibition. The 5-HT antagonists, GR 113808A (5-HT4) at 2.5x10(-6) M and ritanserin (5-HT2) and ondansetron (5-HT3) at 2.5x10(-5) M completely blocked the effect of 5-HT. However, methiothepin (5-HT1) did not produce any effect on serotonin action in the intestinal absorption of amino acid. It can be concluded that 5-HT2, 5-HT3 and 5-HT4 receptors could mediate inhibition of L-leucine absorption across rabbit jejunum. PMID- 9217292 TI - Effect of estradiol on phospholipid lipoprotein levels and fatty acid composition in the rat. AB - Phospholipid content and fatty acid composition in the different serum lipoproteins showed specific variations after castration and estradiol administration. Only the levels of phospholipids in HDL, the principal lipoprotein carrying phospholipids, increased after castration and were further enhanced by estradiol treatment, especially at low doses. Fatty acid composition showed many variations and an irregular pattern. The EFA/NEFA, EFA/ME and SAT/ME ratios were calculated. EFA/ME increased in VLDL after both doses of estradiol, while EFA/NEFA and EFA/ME of LDL enhanced at the low dose and decreased at the high one in a dose-dependent manner. HDL showed higher EFA/ME and SAT/ME ratios after castration and lower values of all ratios after both doses of estradiol. The correlation with diseases more frequent in men is discussed. PMID- 9217293 TI - Sex-specific effects of growth hormone on hepatic 11beta-hydroxysteroid dehydrogenase activity and gene expression in hypothyroid rats. AB - To investigate the effects of growth hormone (GH) on 11beta-HSD1, we determined changes in hepatic 11beta-HSD1 activity in hypothyroid rats following treatment with subcutaneous (s.c) injection of GH for periods ranging from 24 h to 7 days. In male rats, hypothyroidism markedly reduced the hepatic 11beta-HSD1 activity and serum testosterone levels (p < 0.01). Subcutaneous injection of GH once daily to male hypothyroid rats for 48 h inhibited hepatic 11beta-HSD1 activity. However, the same daily dose of GH administered to male hypothyroid rats for 7 days, resulted in a marked increase in hepatic 11beta-HSD1 activity and gene expression (p < 0.01). Furthermore, daily s.c injections of GH to castrated male hypothyroid rats for 7 days reduced hepatic 11beta-HSD1 activity rather than inducing it, the same response seen in hypothyroid female rats. In addition, the treatment of castrated male hypothyroid rats with testosterone for 7 days significantly increased this enzyme activity (p < 0.01). The changes in hepatic 11beta-HSD1 were demonstrated to be associated with the testes in hypothyroid male rats following treatment with GH for 7 days. Moreover, the prolonged exposure to GH required to induce hepatic 11beta-HSD1 in intact hypothyroid male rats and the lack of a similar effect in castrated male hypothyroid rats suggests that this action is indirect and that it may be mediated by androgen production from Leydig cells of the testes and induced by the daily injections of GH. Treatment of hypothyroid male rats with GH at 6-h intervals, however, feminized the hepatic 11beta-HSD1 gene expression. PMID- 9217294 TI - Role of dihydropyridine sensitive calcium channels in glucose transport in skeletal muscle. AB - Glucose transport in skeletal muscle is a carrier-mediated process activated by insulin and by contractile activity. Since previous evidence suggests a role for calcium influx in the activation of this process, the purpose of this study was to determine if glucose transport is mediated by muscle's voltage dependent (dihydropyridine sensitive) calcium channels. Soleus and extensor digitorum longus (EDL) muscles, isolated from rats, were incubated with the calcium channel blocker nifedipine. Basal glucose transport was decreased in both soleus and EDL by nifedipine. Treatment with nifedipine effectively blocked both insulin and contraction stimulated glucose transport in soleus. Conversely, glucose transport in EDL, although reduced, was still significantly increased over basal by both insulin and contraction, due, perhaps, to a relatively greater number of dihydropyridine receptors in EDL. These results provide evidence that contraction stimulated, as well as insulin stimulated, glucose transport is mediated in-part by dihydropyridine receptors in skeletal muscle. PMID- 9217295 TI - Participation of central kappa-opioid receptor in arrhythmogenesis. AB - The effect of kappa receptors agonists and antagonists was studied in the model of epinephrine induced arrhythmias. Kappa receptor agonists U-50,488 and [D-Ala2] Dynorphin A (1-13) administered I.C.V. potentiate the arrhythmogenic effect of epinephrine. The effect of U-50,488 was completely blocked by kappa receptor antagonist, nor-binaltorphine. Administration of N-cholinergic receptor inhibitor, hexamethonium, prevented pro-arrhythmic effects of U-50,488 and [D Ala2]-Dynorphin A (1-13). The data support the hypothesis that central kappa opioid receptors play an important role in the arrhythmogenesis. PMID- 9217296 TI - Occlusal variables, bruxism and temporomandibular disorders: a clinical and kinesiographic assessment. AB - Seventy-five patients suffering from myofascial pain, headaches and anterior disc displacement were assessed clinically and with a kinesiograph. Twenty-eight asymptomatic dental staff served as a control group. The prevalence of awareness of bruxism was significantly greater in our TMD patients than the controls. Bruxism patients recorded a higher prevalence of incisor dentine wear suggestive of a forward mandible posture. Class II, Division 1 malocclusions formed a significantly higher proportion of the TMD patient group than the controls. Kinesiographic recordings showed that the vertical and lateral components of movement from postural position to intercuspal were significantly greater in the patient group. PMID- 9217297 TI - Hardness and fracture toughness of four commercial visible light-cured composite resin veneering materials. AB - Four commercial visible light (VL)-cured composite resin veneering materials with a dentine shade were examined for their Knoop hardness and fracture toughness. Composite specimens were classified into three groups. The first group was cured by VL only, the second group was cured by VL and post-cured by VL and the third group was cured by VL and post-cured by heat. It became evident that one composite containing four-functional urethane monomer had both hardness and fracture toughness greater than those of the other three composites containing two-functional urethane monomer. The filler content (vol%) in the composite tended to be linearly proportional to both hardness and fracture toughness. Post curing by VL and heat were proven to effectively increase both hardness and fracture toughness of once light-cured composites. These results suggest that the clinical performance (e.g. wear resistance and colour stability) of VL-cured composite resin veneering materials might be improved with the aid of post curing. PMID- 9217298 TI - Experimental occlusal interferences. Part V. Mandibular rotations versus hemimandibular translations. AB - Frontal plane mandibular rotations and corresponding hemimandibular translations were studied in vitro by using direct observations of a human cadaver mandible and in vivo by using the indirect observations of rotational electrognathography. A comparison between the two methods showed that rotational electrognathography erred in measuring the clinically relevant hemimandibular translations resulting from mandibular rotations having a unilateral molar point (simulated occlusal interference) as the pivot of frontal plane torque. In vitro frontal plane rotations about a unilateral mandibular molar tooth (simulated occlusal interference) suggested that the resulting hemimandibular upward translations of the lateral portion of the mandibular condyle, contralateral to the molar tooth, would cause considerable compressive loading of the temporomandibular joint disc. PMID- 9217299 TI - The fracture resistance of dental materials. AB - A novel test procedure was developed that provides the basis for a standard plain strain fracture toughness test to evaluate fracture resistance of resin-based materials. This procedure utilizes a notched disc specimen. Using this technique, a new concept of Torque to initiate fracture (T)' was suggested. Also, fracture resistance can be assessed by means of critical stress intensity factor (K[IC]). Using this method, the fracture resistance of direct restorative materials, resin based inlay/onlays and luting cements can be evaluated. PMID- 9217300 TI - General health factors and denture function in patients with burning mouth syndrome and matched control subjects. AB - A total of 30 denture-wearing patients with burning mouth syndrome (BMS) referred to a Pain Clinic Unit and 26 age- and sex-matched control subjects were examined and compared with respect to general health factors and denture function. The study demonstrated a significantly higher frequency of multiple chronic diseases, psychosocial stress factors, and tenderness/pain in masticatory, neck, shoulder, and suprahyoid muscles in patients with BMS. Denture function differed also between the two groups as patients with BMS had significantly less daily use of dentures, reduced tongue space, incorrect placement of occlusal table and increased vertical dimension. Pain interview with the use of the McGill Pain Questionnaire demonstrated that pain in parts of the body other than the oral cavity were reported more frequently and that the intensity of past pain experiences was not rated higher except for pain in the head in patients with BMS. The results suggested a complex interaction between several general health factors, psychosocial stressors and denture dysfunction in order to explain an idiopathic burning pain in the anterior part of the oral cavity. The existence of demonstrable load factors does not seem to support the suggestion that BMS is primarily a psychogenic disorder. PMID- 9217301 TI - Stress in bone adjacent to dental implants. AB - Finite element analysis (FEA) was employed to assess patterns of stress in bone adjacent to an implant after application of loads through an attached distal extension cantilever. Under all loading conditions, the highest stresses occurred at the distal cervical bone margin adjacent to the cantilever. In clinical studies, this is not consistently the site of the greatest bone changes seen radiographically. This suggests that extrapolation of FEA studies to clinical implantology should be approached with caution until further data become available on both mechanical properties of bone and patterns of bone remodelling induced by defined functional stresses in mandible and maxillae. PMID- 9217302 TI - Sella-Nasion line revisited. AB - The anterior cranial base or the Sella-Nasion (SN) line is often used by orthodontists as a reference line for assessment of dentofacial deformities. Its most important value is its relative stability, practicality and the ease of location of both points Sella and Nasion. Reliability of the SN line as a suitable assessor for exact facial measurements has been discussed before. The intent of this study was to identify unusual SN rotations for subjects acquiring normal and abnormal skeletal profiles and hence correct values associated with SN line, both in vertical and horizontal planes. A random sample of 150 British white school children aged 9-12 years old was selected. Lateral head films of the children were traced and digitized. Point Nasion was joined to posterior nasal spine and also to point Sella, thus producing a Z angle when the posterior nasal spine is extended to anterior nasal spine. Means and standard deviations were calculated. The mean angle S-N-PNS was 38 degrees +/- 5 while the mean of angle N PNS-ANS was 47 degrees +/- 3. Comparison of these angles for patients of the same age group with the mean Z angle can identify rotation of the palatal and Sella Nasion planes. Hence corrections of angular measurements involving Sella point can be performed. PMID- 9217304 TI - Polychlorinated biphenyl congener 153-induced ultrastructural alterations in rat liver: a quantitative study. AB - Alterations in the liver of male and female Sprague-Dawley rats fed PCB congener 153 (2,2',4,4',5,5'-hexachlorobiphenyl) at 0.5, 5, or 50 ppm concentrations in diets for 13 weeks were determined morphometrically. A dose-dependent increase in hepatocyte volume was detected; the cytoplasmic compartment contributed to the increase in cell volume in an overwhelming fashion. Eighty percent and 250% increase in smooth endoplasmic reticulum volume and its surface area in hepatocytes were estimated in animals of both genders from 5- and 50-ppm groups, respectively; the organelle played the largest part in the increase in cytoplasmic volume. Rough endoplasmic reticulum alteration was shown to depend on gender, where the volume per hepatocyte was augmented by 40% and 45% in females of 5- and 50-ppm groups, respectively, however, 30% and 20% decreases in volume of this organelle were noted in males at those congener concentrations. A decrease of 13% in normal mitochondria volume at 50 ppm concentration was observed, which may have been a consequence of a transformation of these mitochondria to abnormal types. Two types of abnormal mitochondria, named Type I and Type II, were defined: the former comprised mitochondria that had cristae which laying parallel to the long axis of the organelle and the latter showed C- or ring-shaped profiles. Data analysis revealed a trend toward an increase in abnormal mitochondria volume in the cells as the congener concentration elevated. In addition, a threefold increase in the volume of lysosomal elements per hepatocyte was noted in 50 ppm PCB-fed rats of both genders. Also, a significant increase in peroxisome volume per cell in female rats was detected at a lower concentration than it was in the male. This study, which is a first ultrastructural quantitative investigation on the effects of a PCB that included many parameters. The methodology, and the data may prove useful to provide better understanding of pathology in the evaluation and regulation of toxic chemicals. PMID- 9217303 TI - Changes in neuroreceptor function of tracheal smooth muscle following acute ozone exposure of guinea pigs. AB - We studied the effect of in vivo ozone inhalation (3 ppm, 2 h) on neuroreceptor function in guinea pig tracheal smooth muscle in vitro and the role of the epithelial layer in this process. Changes in smooth muscle tension after stimulation of the muscarinic- and beta-adrenergic receptor were recorded isometrically and stained tracheal tissue sections were histologically evaluated for changes in the epithelial and smooth muscle layer. Ozone exposure resulted in an increase in maximal contraction following stimulation of the muscarinic receptor, whereas pD2 values remained unchanged. After stimulation of the beta adrenergic receptor no increase in maximal relaxation but only an increase in pD2 value was observed after correction for differences in precontraction level in control- and ozone-exposed situations. Mechanical removal of the epithelial layer resulted in a slight increase of the maximal contraction level after stimulation with methacholine in the control situation, whereas exposure to ozone resulted in a strong decrease of the maximal contraction level under these conditions. Histological stainings showed a slight and focal influx of neutrophilic granulocytes in the epithelial layer, submucosal layer and airway lumen after exposure to ozone. These data support the idea that ozone is able to increase the maximal degree of airway narrowing upon muscarinergic stimulation, i.e. a hyperreactivity response. The results also suggest that functionally altered epithelium plays an important role in the process of ozone-induced hyperreactivity, possibly linked with an early inflammatory response. PMID- 9217305 TI - Airborne particulate matter modulates the production of reactive oxygen species in human polymorphonuclear granulocytes. AB - Causal relationships between airborne particles (especially particulate matter < 10 microm in diameter) and increases in prevalences and symptoms of respiratory diseases have been postulated in many epidemiologic studies. Polymorphonuclear leukocytes (PMN) in the nasal or bronchial epithelium can be exposed to particulate matter (PM) and may upon exposure produce reactive oxygen species (ROS). Release of ROS can result in cellular and tissue damage and initiate or exacerbate inflammation. To elucidate the effect of PM on inflammatory reactions, we exposed human PMN to PM extracts. PM were collected with high volume samplers in two cities, Dusseldorf and Duisburg, in Germany and reflect sites with high traffic and industrial emissions respectively. The collected particles were extracted using water and then dichloromethane, resulting in an aqueous and an organic extract. The production of ROS was determined using luminol-enhanced chemiluminescence (LCL) of resting and zymosan-stimulated PMN. The present study shows that extracts of PM alone significantly stimulated the production and release of ROS in resting PMN. The effects of the PM extracts were inhibited by superoxide dismutase (SOD), catalase and sodium azide (NaN3). Zymosan-induced LCL was, however, diminished by coincubation with PM extracts. The chemical composition is important when considering the effects of particles. Our study shows that only organic substances adsorbed to particles stimulate LCL. SOZ induced LCL is inhibited by both types of extracts, but aqueous extracts have a stronger inhibitory effect. It is at present unclear which substances are responsible for these effects. PMID- 9217306 TI - Modulation of cytochrome P450 by 5,5'-bis-trifluoromethyl-2,2'-dichlorobiphenyl, a unique environmental contaminant. AB - 5,5'-Bis-trifluoromethyl-2,2'-dichlorobiphenyl (5,5'CF3-2,2'PCB) is representative of a unique class of trifluoromethyl polychlorinated biphenyls (CF3-PCBs) found in sediments and fish of Lake Ontario, the Niagara river, and their tributaries. The potential hazard of 5,5'CF3-2,2'PCB was assessed by exposing male Wistar rats to this agent in corn oil at a dose of 1 mg/kg/day or 75 mg/kg/day or corn oil alone (control) by oral intubation for 7 consecutive days. No lethality occurred during the course of exposure. A significant increase in liver weight and liver/body weight ratio and significant decrease in body weight gain were observed following exposure to the high dose of CF3-PCB, relative to control. Exposure to the CF3-PCB also resulted in a dose-related increase in the total hepatic cytochrome P450 content. This was associated with a dose-related increase in the O-deethylation of ethoxycoumarin, an activity which is mediated by several cytochrome P450s and thus provides a general representation of cytochrome P450 status. Specifically, a dose-dependent induction of the cytochrome P450s 1A1 and 1A2 (CYP1A1 and CYP1A2) proteins and their respective activities was observed, with significant induction occurring in both the low and high dose CF3-PCB groups, compared to control. Additionally, CYP2B1 and CYP2B2 proteins and activities were induced following treatment with the high dose of 5,5'CF3-2,2'PCB. Since, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and related compounds selectively induce CYP1A1 and CYP1A2. the results suggest that 5,5'CF3-2,2'PCB has significant dioxin-like activity. Furthermore, 5,5'CF3-2,2'PCB appears to be acting as a mixed-type inducer since phenobarbital like induction of CYP2B1 and 2B2 was also associated with exposure. PMID- 9217307 TI - A small peptide from durum wheat gliadin prevents cell agglutination induced by prolamin-peptides toxic in coeliac disease. AB - A peptide (m.w. 1157.5 Da) able to prevent the agglutination of K562(S) cells induced by the peptic-tryptic prolamine digests of the cereals toxic in coeliac disease (i.e. bread wheat, rye, barley and oat) was characterized as one of the components of the peptic-tryptic digest of durum wheat gliadin. This peptide was synthesized in a high degree of purity with the solid phase method with the Applied Biosystem 431A. An amino acid sequence was identified in the 1157.5 Da peptide as being related to the largest common sequences previously detected in a series of bread wheat toxic peptides by other authors. PMID- 9217308 TI - Cadmium uptake by a human hepatic cell line (WRL-68 cells). AB - A hepatic human cell line (WRL-68 cells) was employed to investigate the uptake of the toxic heavy metal cadmium. Cd accumulation in WRL-68 cells is a time-, temperature- and concentration-dependent process. A rapid initial phase of uptake was followed by a second slower phase. The transport does not require energy and 55% of Cd transport occurs by temperature-insensitive processes, possibly by diffusion. The rest of Cd transport (45%) occurs by temperature-sensitive processes, probably ion channels and carriers, that involve interaction with sulfhydryl groups. The calcium channel blockers nifedipine and verapamil inhibit the uptake of cadmium, with an inhibition of 35% after 30 min incubation with 100 microM verapamil and 10 microM Cd. These data suggest that about one third of the Cd enters WRL-68 cells through the calcium channels. The toxic metals appear to use the transport pathways that exist for biologically essential metals. Our results in human hepatic cells are very similar to those reported in cultured rat hepatocytes. It appears that transport pathways available for Cd uptake are similar and independent of the species of hepatocyte origin. Moreover, the WRL-68 cell line seems to be an excellent in vitro model to study the mechanism of cell damage due to Cd. PMID- 9217309 TI - Effect of fructose on the biochemical toxicity of hydrazine in isolated rat hepatocytes. AB - Rat hepatocyte suspensions were incubated with various concentrations of hydrazine (0, 8, 12, 16, 20 mM) for 1, 2 and 3 h. In some experiments fructose (10 mM) was added either during the preincubation period, or 1 h after the start of hydrazine treatment. In certain experiments in which fructose was added, the glycolytic inhibitor sodium fluoride (3 mM) was also added during the preincubation period. Hepatocytes incubated with hydrazine alone demonstrated both a concentration- and time-dependent loss of cell viability as measured by increased Trypan blue uptake and lactate dehydrogenase (LDH) leakage. These parameters were reduced and delayed by fructose when added either before or 1 h after hydrazine treatment. There was also both a concentration- and time dependent loss of ATP and reduced glutathione (GSH) content with hydrazine treatment. Moreover, fructose caused an initial rapid depletion of ATP but thereafter ATP levels were increased in control hepatocytes. Fructose reduced both the depletion of ATP and GSH in hydrazine- treated hepatocytes. Urea synthesis was inhibited by all concentrations of hydrazine studied but fructose treatment after 1 h did not alter this. This study also demonstrated that fluoride, an enolase inhibitor, abolished the protection against depletion of ATP levels provided by fructose, without affecting cell viability or GSH levels. These findings suggest that the cytotoxicity of hydrazine and its effects on urea synthesis and GSH levels are not a direct result of ATP depletion. The protective effects of fructose against the cytotoxicity may be due to a direct interaction with hydrazine. PMID- 9217310 TI - Prenatal exposure to cypermethrin modulates rat NK cell cytotoxic functions. AB - The synthetic pyrethroid insecticide, cypermethrin, was given during gestation to pregnant rats by gavage in corn oil. Peripheral blood and spleen cytotoxic activity of dams and their offspring were then evaluated at different times (30, 60, 90, 120 days) after birth. Pups showed a significant increase in peripheral blood natural killer (NK) and antibody-dependent (ADCC) cytotoxic activity paralleled with a similar increase in the percentage of NK-RP1+ cells and decreased activity in the spleen. Pregnant cypermethrin-exposed dams showed no changes in peripheral blood or spleen cytotoxic function during the postnatal period. Overall, these results suggest that immunomodulation of cytotoxic activity observed in the offspring is likely attributable to a specific effect of cypermethrin administered during the prenatal period. PMID- 9217311 TI - Toxicology of blast overpressure. AB - Blast overpressure (BOP) or high energy impulse noise, is the sharp instantaneous rise in ambient atmospheric pressure resulting from explosive detonation or firing of weapons. Blasts that were once confined to military and to a lesser extent, occupational settings, are becoming more universal as the civilian population is now increasingly at risk of exposure to BOP from terrorist bombings that are occurring worldwide with greater frequency. Exposure to incident BOP waves can cause auditory and non-auditory damage. The primary targets for BOP damage are the hollow organs, ear, lung and gastrointestinal tract. In addition, solid organs such as heart, spleen and brain can also be injured upon exposure. However, the lung is more sensitive to damage and its injury can lead to death. The pathophysiological responses, and mortality have been extensively studied, but little attention, was given to the biochemical manifestations, and molecular mechanism(s) of injury. The injury from BOP has been, generally, attributed to its external physical impact on the body causing internal mechanical damage. However, a new hypothesis has been proposed based on experiments conducted in the Department of Respiratory Research, Walter Reed Army Institute of Research, and later in the Department of Occupational Health, University of Pittsburgh. This hypothesis suggests that subtle biochemical changes namely, free radical-mediated oxidative stress occur and contribute to BOP-induced injury. Understanding the etiology of these changes may shed new light on the molecular mechanism(s) of injury, and can potentially offer new strategies for treatment. In this symposium. BOP research involving auditory, non-auditory, physiological, pathological, behavioral, and biochemical manifestations as well as predictive modeling and current treatment modalities of BOP-induced injury are discussed. PMID- 9217312 TI - The pathology of primary blast overpressure injury. AB - Primary blast injury occurs in civilian and military detonations and from the firing of weapon systems. The pathology of primary blast injury has been reported for the last 70 years and has primarily been limited to descriptions of gross pathology and histology. Commonly accepted tenets have not been confirmed as blast overpressure experiments in enclosures and with multiple detonations have been conducted. Organ systems other than the ear and the lung are playing a greater role in injury definition and research importance. This paper is an overview and update of the current understanding of the pathology of primary blast injury. PMID- 9217313 TI - Blast overpressure induced structural and functional changes in the auditory system. AB - Blast overpressure of sufficient intensity can produce injury to various organ systems. Unprotected ears result in the auditory system being the most susceptible. The injuries to the auditory system include: rupture of the tympanic membrane, dislocation or fracture of the ossicular chain, and damage to the sensory structures on the basilar membrane. All these injuries can be characterized as a form of mechanical damage to the affected structure. Injury to the sensory structures on the basilar membrane leads to temporary and permanent loss of hearing sensitivity. The temporary component of the hearing loss shows a time course after removal from the noise which frequently will include an initial increase in hearing loss followed by a recovery period during which threshold may return to preexposure levels or stabilize at a higher level which represents a permanent loss of hearing sensitivity. This type of recovery function suggests that there are damage processes which continue after the traumatic event and that intervention might mitigate some of the damage and hearing loss. PMID- 9217315 TI - Maximal exercise performance-impairing effects of simulated blast overpressure in sheep. AB - Lung contusion has been identified as a primary blast injury. These experiments addressed a fundamental and overt endpoint of primary blast injury, incapacitation (performance decrement). Respiration, hemodynamics, and blood gases were measured in sheep undergoing incremental exercise challenge before and 1 h after simulated blast exposure of the thorax. Pathologic examination of lung tissue was performed after exposure and exercise testing. Blast overpressure was simulated in the laboratory using a compressed air-driven shock tube. Three levels of lung injury (Levels 1-3, 'Trivial', 'Slight', and 'Moderate' injury, respectively) were examined for effects on maximal oxygen consumption (VO[2max]), an index of cardiorespiratory fitness. Resting hemodynamics and blood gases were relatively normal an hour after exposure, immediately before exercise. However, Levels 1-3 lung injury were associated with average 4.8, 29.9 and 49.3% VO(2max). decreases, respectively. These performance decrements for Levels 2 and 3 were significantly different from respective controls (non-exposed). Exercise caused significant hemoconcentration in sheep under control conditions, before exposure (resting 9.5 +/- 0.9, end-exercise 11.8 +/- 0.9 g/100 ml). Blast exposure resulted in average decreases of 4.9 +/- 3.4, 12.8 +/- 4.0, and 12.6 +/- 3.3% in exercise-induced hemoconcentration for Levels 1-3 injury, respectively. Normal exercise-induced hemodynamic increases were also attenuated after exposure. Levels 2 and 3 injury resulted in average 22.6 +/- 2.9 and 18.5 +/- 11.2% stroke volume decreases, and also 22.3 +/- 8.4 and 29.0 +/- 14.2% cardiac output decreases, respectively, during exercise. While blast lung pathology and pulmonary function changes could account for post-blast performance decrements, these experiments suggest that in sheep, early after exposure, diminished hemoconcentration and cardiac disfunction may also contribute to decreased exercise performance. PMID- 9217314 TI - Visual system degeneration induced by blast overpressure. AB - The effect of blast overpressure on visual system pathology was studied in 14 male Sprague-Dawley rats weighing 360-432 g. Blast overpressure was simulated using a compressed-air driven shock tube, with the aim of studying a range of overpressures causing sublethal injury. Neither control (unexposed) rats nor rats exposed to 83 kiloPascals (kPa) overpressure showed evidence of visual system pathology. Neurological injury to brain visual pathways was observed in male rats surviving blast overpressure exposures of 104-110 kPa and 129-173 kPa. Optic nerve fiber degeneration was ipsilateral to the blast pressure wave. The optic chiasm contained small numbers of degenerated fibers. Optic tract fiber degeneration was present bilaterally, but was predominantly ipsilateral. Optic tract fiber degeneration was followed to nuclear groups at the level of the midbrain, midbrain-diencephalic junction, and the thalamus where degenerated fibers arborized among the neurons of: (i) the superior colliculus, (ii) pretectal region, and (iii) the lateral geniculate body. The superior colliculus contained fiber degeneration localized principally to two superficial layers (i) the stratum opticum (layer III) and (ii) stratum cinereum (layer II). The pretectal area contained degenerated fibers which were widespread in (i) the nucleus of the optic tract, (ii) olivary pretectal nucleus, (iii) anterior pretectal nucleus, and (iv) the posterior pretectal nucleus. Degenerated fibers in the lateral geniculate body were not universally distributed. They appeared to arborize among neurons of the dorsal and ventral nuclei: the ventral lateral geniculate nucleus (parvocellular and magnocellular parts); and the dorsal lateral geniculate nucleus. The axonopathy observed in the central visual pathways and nuclei of the rat brain are consistent with the presence of blast overpressure induced injury to the retina. The orbital cavities of the human skull contain frontally-directed eyeballs for binocular vision. Humans looking directly into an oncoming blast wave place both eyes at risk. With bilateral visual system injury, neurological deficits may include loss or impairments of ocular movements, and of the pupillary and accommodation reflexes, retinal hemorrhages, scotomas, and general blindness. These findings suggest that the retina should be investigated for the presence of traumatic or ischemic cellular injury, hemorrhages, scotomas, and retinal detachment. PMID- 9217316 TI - Exposure to sublethal blast overpressure reduces the food intake and exercise performance of rats. AB - Exposure to blast overpressure can typically inflict generalized damage on major organ systems, especially gas-containing organs such as the lungs and the gastrointestinal tract. The purpose of the present study was to use rat's food intake and exercise wheel running as behavioral correlates of the perhaps more subtle damage to these organ systems induced by sublethal blast overpressure. Toward this end, all rats were exposed to a 12-h light/dark cycle and food was available only in the dark period. Prior to exposure, rats in the (E)xercise group were required to execute five rotations of an activity wheel for a food pellet; wheel turns that occurred at times other than when a rat was feeding were recorded separately and labeled exercise running. In the (S)edentary and (A)nesthesia groups, wheel running was not possible and rats were required to execute five leverpresses for a single pellet. A compressed air-driven shock tube was used to expose rats to a supra-atmospheric wave of air pressure. The tube was separated into two sections by a polyester membrane, the thickness of which determined peak and duration of overpressure. All rats were anesthetized with 50 mg/kg of phenobarbital. After reaching a deep plane of anesthesia, they were individually tied in a stockinet across one end of the shock tube. In preliminary tests, the membrane thickness was 1000 (A)ngstroms and rats in Group L(ethality) were exposed to a 129 kPa (peak amplitude) wave of overpressure. Three of six rats survived exposure to this peak pressure; pathology was evident in the lungs and gastrointestinal tract of all non-survivors. Rats in Groups E and S were tested with a 500 A membrane, which resulted in an 83 kPa peak amplitude. All rats survived exposure to this lower peak pressure. On the day of exposure to blast, the relative reduction of intake during the first 3 h of the dark period was significantly greater for Group E than for Groups S and A; the intake of Groups E and S remained reduced for four additional recovery days. Bodyweight was not significantly affected. Exercise wheel running also was reduced significantly on the day of exposure and during subsequent recovery days. These preliminary findings suggest that exposure to sublethal blast overpressure can reduce food consumption and exercise performance, perhaps as a consequence of damage to the gastrointestinal tract and lungs. PMID- 9217317 TI - A proposed biochemical mechanism involving hemoglobin for blast overpressure induced injury. AB - Blast overpressure (BOP) is the abrupt, rapid, rise in atmospheric pressure resulting from explosive detonation, firing of large-caliber weapons, and accidental occupational explosions. Exposure to incident BOP waves causes internal injuries, mostly to the hollow organs, particularly the ears, lungs and gastrointestinal tract. BOP-induced injury used to be considered of military concern because it occurred mostly in military environments during military actions or training, and to a lesser extent during civilian occupational accidents. However, in recent years with the proliferation of indiscriminate terrorist bombings worldwide involving civilians, blast injury has become a societal concern, and the need to understand the biochemical and molecular mechanism(s) of injury, and to find new and effective methods for treatment gained importance. In general, past BOP research has focused on the physiological and pathological manifestations of incapacitation, thresholds of safety, and on predictive modeling. However, we have been studying the molecular mechanism of BOP-induced injury, and recently began to have an insight into that mechanism, and recognize the role of hemoglobin released during hemorrhage in catalyzing free radical reactions leading to oxidative stress. In this report we discuss the biochemical changes observed after BOP exposure in rat blood and lung tissue, and propose a biochemical mechanism for free radical-induced oxidative stress that can potentially complicate the injury. Moreover, we observed that some antioxidants can interact with Hb oxidation products (oxy-, met- and oxoferrylHb) and act as prooxidants that can increase the damage rather than decrease it. PMID- 9217318 TI - Biological response to blast overpressure: a summary of modeling. AB - A soldier in training is exposed to a variety of blast sources that can adversely affect his auditory and nonauditory systems. While auditory standards have been formulated for many decades, knowledge about nonauditory effects of blast have not been captured in a criterion that can be applied to all circumstances. For the past 15 years, JAYCOR, working together with the Walter Reed Army Institute of Research, has been using modeling, simulation, and data analysis to determine the nature of injury in animal models, capture that understanding in physiologically correct mathematical models, and extend the findings to objective criteria that can be used to set exposure limits. This paper summarizes the accomplishments of that effort. PMID- 9217319 TI - Management of primary blast injury. AB - Blast waves are produced following the detonation of munitions, the firing of large caliber guns, or from any type of explosion. These blast waves can be powerful enough to injure the individuals exposed to them. This type of injury is called primary blast injury (PBI) and the organs most vulnerable to PBI are the gas-filled organs, namely the ear, the lungs and the gastrointestinal tract. The approach to the casualty with PBI is the same as it would be for any trauma victim, i.e. the initiation of life support measures. Attention should be directed to the common life-threatening manifestation of thoracic and abdominal PBI. Pulmonary manifestations would include hemorrhage, barotrauma and arterial air embolism, while abdominal manifestations would include hemorrhage and hollow organ rupture. Therapy is directed at the specific manifestations as well as avoiding additional iatrogenic injury. PMID- 9217320 TI - When helicase and topoisomerase meet! AB - Several examples of direct interactions between helicases and topoisomerases have recently been described. The data suggest a possible cooperation between these enzymes in major DNA events such as the progression of a replication fork, segregation of newly replicated chromosomes, disruption of nucleosomal structure, DNA supercoiling, and finally recombination, repair, and genomic stability. A first example is the finding of a strong interaction between T antigen and topoisomerase I in mammalian cells, that may trigger unwinding of the parental DNA strands at the replication forks of Simian Virus 40. A second example is the reverse gyrase from thermophilic prokaryotes, composed of a putative helicase domain, and a topoisomerase domain in the same polypeptide. This enzyme may be required to maintain genomic stability at high temperature. A third example is the finding of an interaction between type II topoisomerase and the helicase Sgs1 in yeast. This interaction possibly allows the faithful segregation of newly replicated chromosomes in eukaryotic cells. A fourth example is the interaction between the same helicase Sgs1 and topoisomerase III in yeast, that may control recombination level and genetic stability of repetitive sequences. Recently, in humans, mutations in genes similar to Sgs1 have been found to be responsible for Bloom's and Werner's syndromes. The cooperation between helicases and topoisomerases is likely to be extended to many aspects of DNA mechanisms including chromatin condensation/decondensation. PMID- 9217321 TI - Cartilage associated protein (CASP) is a novel developmentally regulated chick embryo protein. AB - A subtracted cDNA library was generated to identify cDNAs specific for chondrocyte mRNAs preferentially expressed at the hypertrophic stage with respect to early differentiation stages. The characterization of a cDNA isolated from this library that hybridizes with a 1.8 kb mRNA is described here. This mRNA is expressed at extremely low levels in dedifferentiated chondrocytes cultured in adherent conditions, at very low levels in differentiating chondrocytes and at very high levels in hypertrophic chondrocytes cultured in suspension conditions. In the developing chick embryo this mRNA is detectable in RNAs extracted from several other tissues besides cartilage. The described cDNA contains a complete open reading frame coding for a polypeptide of about 33 kDa. Homology searches with known cDNA and protein sequences have revealed that the chicken protein is related to the amino-terminal half of two mammalian nuclear antigens. By immunohistochemistry with specific rabbit antisera a strong signal was detected in the cartilage extracellular matrix of selected regions of the developing skeleton. Because of this localization of the antigen we named this protein cartilage associated protein (hereafter referred to as CASP). PMID- 9217322 TI - Ligand recruitment by vinculin domains in transfected cells. AB - Vinculin, a prominent protein component of microfilament-membrane attachment sites, consists of three major domains: an N-terminal, compact head and a C terminal rod-like tail that are connected by a flexible, proline-rich hinge. In vitro, the protein has been shown to interact with numerous ligands, including other components of the microfilament system. To characterize the ligand recruitment ability of the different vinculin domains in a cellular environment, we used a novel approach of comprising chimeric proteins of either the vinculin head, hinge or tail regions, fused to the membrane anchor sequence of ActA, a surface protein of the intracellular bacterial pathogen Listeria monocytogenes. When PtK2 cells were transfected with the corresponding constructs, the ActA membrane anchor directed the chimeric polypeptides to mitochondrial membranes. In this position, they accumulated microfilament proteins, as seen by immunofluorescence analysis. A chimera comprising the full length vinculin clone recruited a substantial amount of the cellular F-actin, the vasodilator stimulated phosphoprotein (VASP) and paxillin, but little alpha-actinin and talin. The presence of only the vinculin head directed some of the fusion protein to focal contacts, and alpha-actinin recruitment was still ineffective. Prominent recruitment of F-actin and of VASP required the presence of the tail and proline rich hinge, respectively. Reducing the vinculin tail to short pieces harboring only one of the two F-actin binding sequences, which were defined by in vitro experiments, resulted in loss of activity, possibly by incorrect polypeptide folding. The proline-rich hinge domain could be exchanged for the analogous region of the ActA protein, and the number of such proline-clusters, containing an FPPPP motif, correlated with the extent of VASP recruitment. The results show that this system can be used to analyze in vivo the activity of vinculin domains responsible for the assembly of various cytoskeletal ligands. PMID- 9217324 TI - Calcium ion dependency and the role of inositol phosphates in melatonin-induced encystment of dinoflagellates. AB - The unicellular eukaryotic dinoflagellates shed their flagella and form a new pellicle cyst wall in response to environmental stress. This encystment process can also be induced by indoleamines such as melatonin and 5-methoxytryptamine. To decipher the complex signaling events which lead to encystment, we have investigated the functional roles of Ca2+ and inositol phosphates in indoleamine induced encystment of the dinoflagellates Alexandrium catenella and Crypthecodinium cohnii. Pretreatment with EGTA, but not with EDTA, effectively blocked the indoleamine-induced encystment of A. catenella in a dose-dependent manner. Conversely, agents that facilitate the influx of Ca2+ (Bay K 8644, A23187 and ionomycin) dose-dependently induced encystment of A. catenella. Endoplasmic Ca2+-ATPase inhibitors such as thapsigargin and the peptide toxin melittin also induced encystment of A. catenella. These results suggest that an elevation of intracellular [Ca2+] may be involved in the encystment response. In terms of the regulation of phospholipase C, melatonin dose- and time-dependently stimulated the formation of inositol phosphates in C. cohnii. The rank order of potency for several indoleamines to stimulate inositol phosphates formation was 2 iodomelatonin > 5-methoxytryptamine > or = melatonin >> N-acetylserotonin > 5 hydroxytryptamine. This rank order was the same as for the indoleamine-induced encystment of C. cohnii as previously reported. Our results indicate that indoleamine-induced activation of phospholipase C and elevation of intracellular [Ca2+] may be proximal steps in the signal transduction pathway leading to encystment in dinoflagellates. Moreover, this is the first demonstration of the possible involvement of Ca2+ and inositol phosphates as second messengers in dinoflagellates. PMID- 9217323 TI - Microtubule-entrained kinase activities associated with the cortical cytoskeleton during cytokinesis. AB - Research over the past few years has demonstrated the central role of protein phosphorylation in regulating mitosis and the cell cycle. However, little is known about how the mechanisms regulating the entry into mitosis contribute to the positional and temporal regulation of the actomyosin-based contractile ring formed during cytokinesis. Recent studies implicate p34cdc2 as a negative regulator of myosin II activity, suggesting a link between the mitotic cycle and cytokinesis. In an effort to study the relationship between protein phosphorylation and cytokinesis, we examined the in vivo and in vitro phosphorylation of actin-associated cortical cytoskeletal (CSK) proteins in an isolated model of the sea urchin egg cortex. Examination of cortices derived from eggs or zygotes labeled with 32P-orthophosphate reveals a number of cortex associated phosphorylated proteins, including polypeptides of 20, 43 and 66 kDa. These three major phosphoproteins are also detected when isolated cortices are incubated with [32P]ATP in vitro, suggesting that the kinases that phosphorylate these substrates are also specifically associated with the cortex. The kinase activities in vivo and in vitro are stimulated by fertilization and display cell cycle-dependent activities. Gel autophosphorylation assays, kinase assays and immunoblot analysis reveal the presence of p34cdc2 as well as members of the mitogen-activated protein kinase family, whose activities in the CSK peak at cell division. Nocodazole, which inhibits microtubule formation and thus blocks cytokinesis, significantly delays the time of peak cortical protein phosphorylation as well as the peak in whole-cell histone H1 kinase activity. These results suggest that a key element regulating cortical contraction during cytokinesis is the timing of protein kinase activities associated with the cortical cytoskeleton that is in turn regulated by the mitotic apparatus. PMID- 9217325 TI - Receptor-mediated endocytosis of urokinase-type plasminogen activator is regulated by cAMP-dependent protein kinase. AB - Internalization of the urokinase-type plasminogen activator (uPA) requires two receptors, the uPA receptor (uPAR) and the low density lipoprotein receptor related protein (LRP)/alpha2-macroglobulin (alpha2M) receptor. Here, we address whether protein kinases are involved in the internalization of uPA by human melanoma cells. Initially, we found that the internalization of uPA was significantly inhibited by the serine/threonine protein kinase inhibitors staurosporine, K-252a and H-89, but not by the tyrosine kinase inhibitors, genistein and lavendustin A. Internalization of uPA was also inhibited by a pseudosubstrate peptide for cAMP-dependent protein kinase (PKA), but not by a pseudosubstrate peptide for protein kinase C. We confirmed a requirement for PKA activity and implicated a specific isoform by using an antisense oligonucleotide against the regulatory subunit RI alpha of PKA which suppresses PKA-I activity. Exposure of cells to this oligonucleotide led to a specific, dose-dependent decrease in RI alpha protein and to a significant inhibition in the rate of uPA internalization. We further demonstrate that treatment of melanoma cells with either H-89 or PKA RI alpha antisense oligonucleotides also resulted in a decreased internalization of two other ligands of LRP, activated alpha2M and lactoferrin, indicating that PKA activity is associated with LRP. Finally, we demonstrate that PKA activity is also required for the internalization of transferrin, but not for the internalization of the epidermal growth factor or adenovirus 2, suggesting that in melanoma cells, PKA activity is not generally required for clathrin-mediated endocytosis, but is rather associated with specific internalization receptors. PMID- 9217326 TI - Crystalline mitochondrial inclusion bodies isolated from creatine depleted rat soleus muscle. AB - Rats were fed a 2% guanidino propionic acid diet for up to 18 weeks to induce cellular creatine depletion by inhibition of creatine uptake by this creatine analogue. Ultrastructural analysis of creatine depleted tissues showed that mitochondrial intermembrane inclusion bodies appeared in all skeletal muscles analysed, after 11 weeks of feeding. Heart had relatively few even after 18 weeks of analogue feeding and none were evident in kidney, brain or liver. These structures were strongly immuno-positive for sarcomeric mitochondrial creatine kinase and upon removal from mitochondria, the inclusion bodies were shown to diffract to a resolution of 2.5 nm. Two-dimensional image analysis and three dimensional reconstruction revealed arrays of creatine kinase octamers with additional components between the octameric structures. The same mitochondria had a 3-fold higher extractable specific creatine kinase activity than controls. Molecular mass gel filtration of inclusion body containing mitochondrial extracts from analogue fed rat solei revealed mitochondrial creatine kinase eluting as an aggregate of an apparent molecular mass > or = 2,000 kDa. Mitochondrial creatine kinase of control soleus mitochondrial extract eluted as an octamer, with a molecular mass of 340 kDa. Respiration measurements of control solei mitochondria displayed creatine mediated stimulation of oxidative phosphorylation that was absent in analogue-fed rat solei mitochondria. The latter also had 19% and 14% slower rates of state 4 and maximal state 3 respiration, respectively, than control mitochondria. These results indicate that mitochondrial creatine kinase co-crystallises with another component within the inter membrane space of select mitochondria in creatine depleted skeletal muscle, and is inactive in situ. PMID- 9217327 TI - Glycosaminoglycans modulate fibronectin matrix assembly and are essential for matrix incorporation of tenascin-C. AB - We have investigated the role of glycosaminoglycans in fibronectin matrix assembly and the incorporation of tenascin-C into matrix fibrils. Chinese hamster ovary cell mutants with a total block in heparan and chondroitin sulfate production failed to assemble a fibronectin matrix, and incorporated no tenascin C. Another mutant with reduced heparan sulfate produced a normal fibronectin matrix but failed to incorporate tenascin-C. Excess soluble glycosaminoglycans inhibited the binding of tenascin-C to purified fibronectin in ELISA, and completely blocked incorporation into matrix fibrils. Treating cultured cells with xyloside, which interferes with glycosaminoglycan attachment to proteoglycans, also completely blocked their ability to incorporate tenascin-C into matrix fibrils. We conclude that proteoglycans bound to fibronectin fibrils play a major role in binding tenascin-C to these fibrils. We examined more closely the large heparan sulfate proteoglycan, perlecan, and found that it co localizes with tenascin-C and fibronectin in the matrix. The perlecan binding site in tenascin-C was mapped to the fibronectin type III domains 3-5, but this binding was strongly enhanced for the small splice variant, which is the major form incorporated into the matrix. Apparently when the alternative splice segment is inserted after domain 5 it inhibits perlecan binding. Thus heparan sulfate glycosaminoglycans, and perlecan in particular, may play a role in incorporation of the small splice variant of tenascin-C into fibronectin matrix fibrils. PMID- 9217328 TI - NPXY motifs control the recruitment of the alpha5beta1 integrin in focal adhesions independently of the association of talin with the beta1 chain. AB - With the exception of the divergent beta4 and beta8 chains, the integrin beta subunit cytoplasmic domains are short and highly conserved sequences. Consensus motifs are found among the different cytoplasmic beta chains. Experiments using chimeric receptors demonstrated that the 47 amino acids of the beta1 subunit cytoplasmic domain contain sufficient information to target integrins to adhesion plaques. Three clusters of amino acids, named cyto-1, cyto-2 and cyto-3, seem to contribute to this localization. Cyto-2 and cyto-3 exhibit NPXY motifs. At present, the exact function of these motifs remains unknown but it is likely that these sequences are involved in protein-protein interactions. Although NPXY motifs often act as internalization signals at the cytoplasmic tail of membrane receptors, our previous results showed that the two NPXY motifs are not responsible for the alpha5beta1 integrin endocytosis. Herein, we address the question of the role of the two highly conserved NPXY motifs found in the beta1 cytoplasmic domain, and which correspond to the conserved domains cyto-2 and cyto 3. We demonstrate that, within the integrin beta1 cytoplasmic tail, the two NPXY motifs are required for the recruitment of the integrin in focal adhesions. In addition, our results indicate that these two motifs control but do not belong to the talin-binding sites. Finally, the analysis of the phenotypes of NPXY mutants reveals that the interaction of talin with the beta1 cytosolic domain is not sufficient to target the integrins to focal adhesions. PMID- 9217329 TI - The effects of state intergovernmental transfers on local public mental health services. AB - The provision of public mental health services is shifting increasingly from states to local areas. Yet state governments continue to bear financial responsibility for the majority of these services. One implication of this trend is that the success of state policies become dependent on a state's ability to influence the behavior of local areas. This paper discusses the different options states have in designing intergovernmental grant contracts with local areas, and describes likely impacts of the different strategies. These propositions are then tested using data from the Ohio state mental health system from 1989-1993. This study finds that the design of grants affects public expenditures, local revenue generation, and the mix of services provided at the local level. PMID- 9217330 TI - Carving-out mental health benefits to Medicaid beneficiaries: a shift toward managed care. AB - Several states have designed and implemented innovative programs for Medicaid beneficiaries that carve-out the provision of mental health from general health care. This paper describes several such programs and outlines the choices states face in designing these services. Major decisions include the selection of a public or private agency, how that agency is chosen, reimbursement schemes, eligibility criteria, and benefits to be covered. While carve-out programs have yielded initial savings, more research is needed on their effect on quality of care and general health care costs. PMID- 9217331 TI - Contracting between public and private providers: a survey of mental health services in California. AB - This paper reports on a public authority's decision to "make" or "buy" mental health services. Data come from key informant interviews with California county contract or program managers. The questionnaire measures the extent of contracting and the importance of factors that are hypothesized to affect the relative costs of contracting. The percent of contracting by programs ranges from zero to 100, averaging 41%. Sixty-two percent of rural programs perceive little or no competition for public mental health contracts, and contract significantly less than urban programs. The extent of contracting is related to economic and public organizational factors. PMID- 9217332 TI - How well do ambulatory care groups predict expenditures on mental health and substance abuse patients? AB - This paper evaluates the ability of Ambulatory Care Groups (ACGs) to prospectively predict mental health and substance abuse expenditures and total health care expenditures of persons enrolled in the New Hampshire Medicaid Program during fiscal years 1993 and 1994. A series of multi-part models is estimated separately for adults and children and a synthetic R-squared and the mean absolute predictive error are calculated. The results show that with the exception of predicting total expenditures for children, ACGs do not perform as well as simple models containing various demographic and prior mental health/substance abuse utilization measures. PMID- 9217333 TI - Make or buy: HMOs' contracting arrangements for mental health care. AB - Many health maintenance organizations (HMOs) are contracting with external vendors for mental health care, rather than maintaining an internal mental health department. We develop a framework for analyzing HMOs' contracting choices, rooted in transaction cost economics. Applying this framework, external contracting seems most likely to appeal to smaller, newer HMOs and those located in areas with multiple vendors. Pressure from value-oriented buyers may make it harder for HMOs to provide mental health internally, without costly reforms to their product. HMO contracting arrangements deserve further study, given their implications for cost and the quality of care. PMID- 9217334 TI - 'Le roi le veult'. AB - Following the fiftieth anniversary of the National Health Service Act 1946, I make the case in this article that professional provision of health care must remain the property of the citizen under common law and must not become a commodity that is bought and sold by third parties, including the state. PMID- 9217335 TI - Child dental care. PMID- 9217336 TI - Latex allergy. PMID- 9217337 TI - The future of dental amalgam: a review of the literature. Part 5: Mercury in the urine, blood and body organs from amalgam fillings. AB - This is the fifth article in a series of seven on the future of dental amalgam. This covers the studies of mercury distribution to the blood, body organs and the fetus and its excretion in the urine and faeces of humans and experimental animals. It firstly describes the clinical studies comparing the blood and urine mercury levels in patients with and without amalgam fillings and goes on to consider attempts which have been made to calculate tolerable mercury thresholds for the urine. It secondly describes the studies on the body distribution of mercury from amalgam restorations in experimental animals and human cadavers. It finally describes the studies of mercury distribution to the fetus during pregnancy and includes both studies of experimental animals and human clinical studies. The factors affecting the accuracy of these calculations and the relevance of these results is also extensively discussed. PMID- 9217338 TI - An assessment of capitation in the General Dental Service Contract. 1. The level of caries and its treatment in regularly attending children and adolescents. AB - OBJECTIVE: To assess the dental health of regularly attending 7-8- and 14-15-year olds registered under capitation in 1994. To compare the dental health of regularly attending 14-15-year-olds registered under capitation in 1994 with regularly attending patients of a similar age treated under fee-for-service in 1989. DESIGN: Random samples of 7-8- and 14-15-year-olds. Data were recorded on decayed and filled teeth, extracted for caries and teeth fissure sealed. RESULTS: Prevalence of caries in first permanent molars of 7-8-year-olds was 15-16%. Mean caries experience of deciduous posterior teeth was 1.78-2.51. 83-86% had no more than two untreated, decayed, posterior deciduous teeth. Prevalence of caries in 14-15-year-olds was 44-60% while mean caries experience was 1.29-1.83. Reductions in caries experience of 30-39% in 14-15-year-olds for 1989-1994 were due mainly to falls of 42-45% in mean numbers of teeth filled. Increases in mean numbers of decayed, untreated teeth were 0.07-0.11. The proportion of patients with teeth extracted because of caries was 2.4-3.8%. 30-50% had fissure sealants in 1994 compared with 13-25% in 1989. CONCLUSIONS: Dental health of regularly attending capitation patients is generally satisfactory with little evidence of 'supervised neglect'. Prevalence of fissure sealants has increased while the numbers of filled teeth has reduced. Numbers of decayed, untreated teeth have increased but the numbers of teeth extracted for caries have remained low. PMID- 9217339 TI - The use by general dental practitioners of an oral pathology diagnostic service over a 20-year period: the Birmingham Dental Hospital experience. AB - OBJECTIVE: To investigate the use of an oral pathology service by general dental practitioners over a 20-year period. DESIGN: Retrospective analysis of the number of cases received for histological examination in 1975, 1984 and 1994. SETTING: Birmingham Dental Hospital. SUBJECTS AND METHODS: 1101, 2395 and 3366 specimens were accessed respectively for the three years studied. Number and proportion of specimens received, number and proportion that were hard or soft tissue specimens and the information on the request form were recorded. A comparison was made between the provisional clinical and histological diagnoses. RESULTS: Although the number of specimens accessed increased approximately 3-fold, the number of specimens accessed from general dental practitioners increased 5-fold. The variety of histological categories increased by over 50%, most being soft tissue specimens. The number of correct provisional diagnoses increased steadily but the percentage with inappropriate provisional diagnoses remained the same. Information on request forms steadily improved. CONCLUSIONS: The increased number of specimens received from general dental practitioners over the 20-year period reflects an increased demand for an oral pathology diagnostic service. The referral pattern most likely indicates an increased awareness by general dental practitioners of the need to biopsy lesions arising within the oral cavity. PMID- 9217340 TI - Subdural empyema resulting from displacement of a root into the maxillary antrum. AB - Subdural empyema is a rare but serious complication of paranasal sinusitis which may result in death or permanent disability in a significant proportion of cases. A case is presented in which displacement of a premolar root in the maxillary sinus led to a subdural empyema and a resultant left-sided hemiplegia. This case illustrates the necessity of early surgical intervention to remove displaced roots in the maxillary antrum in order to prevent the serious complications of paranasal sinusitis. PMID- 9217341 TI - Dental futurescope. PMID- 9217342 TI - The introduction of dental team education. AB - An initial experiment of introducing trainee dental nurses to assist dental undergraduates for one session a week has led to the introduction of dental team education within the department of restorative dentistry, University of Sheffield. At the same time, self-directed and problem based learning was introduced for the team. This article outlines the development and results of the exercise. PMID- 9217343 TI - Interactive psychoeducational group therapy for traumatized women. AB - A specialized form of outpatient group therapy with traumatized women is described. Interactive psychoeducational group therapy (IPGT) aims to help the survivor differentiate her self-representations from traumatic schemata that she may have assimilated since the traumatic event. Such assimilation is viewed as leading to a number of negative effects, including shame, social isolation, distorted body image, and sense of meaninglessness. Using a membership with heterogeneous trauma, cognitive-distancing techniques, corrective interpersonal enactments, and specifically designed ceremonies, IPGT attempts to encourage survivors to alter their relationship to the traumatic event and the illness of posttraumatic stress disorder. Clinical examples demonstrate members' improvement in self-image, interpersonal relationships, and sense of belonging to the community at large. PMID- 9217344 TI - Predicting patient benefit from a group-oriented, evening treatment program. AB - Patients with coexisting mood and personality disorders are a challenge to service care providers who offer treatment. The response of 190 patients with one or both disorders to an intensive group-oriented evening treatment program was investigated. Benefit was assessed using factors derived from pre-post outcome measures and from general impressions of program usefulness. A predictive model consisting of patient levels of psychological mindedness and psychodynamic work was applied. Results indicated that psychological mindedness was significantly related to psychodynamic work in the program, and work was related to general impressions of program usefulness. These results cross-validated findings from a previous study of a day-treatment program. Restricted range may have prevented finding significant results for the pre-post outcome factors. Clinical implications of the findings are discussed. PMID- 9217345 TI - An object relations perspective on couples group therapy. AB - This article presents a couples group therapy treatment approach that uses analytic object relations concepts such as transference/countertransference, projective identification, containment, and the holding environment. Object relations theory is seen as the most useful theory with which to view couple interaction because it is based on a two-person psychology and focuses on the impact of relational systems on the development of the person. This includes the idea that the person grows within the attachment to another person. Group therapy is seen as a more effective treatment approach because the group is a resilient holding environment that provides an avenue in which projective identifications can be understood and contained, power can be redefined, isolation of the couple can be decreased, and the couple's responses can become more versatile. The model described is illustrated with clinical vignettes from an open-ended couples group. PMID- 9217346 TI - The pivotal group member: a study of treatment-destructive resistance in group therapy. AB - This article defines the pivotal group member (PGM) and highlights the differences between the PGM and other central figures in group therapy. Whereas central figures adopt predictable roles that serve therapeutic functions, the PGM produces a treatment-destructive atmosphere within the group, one that shrouds the group in feelings of heaviness and hopelessness. The PGM dynamic can only occur in a three-way collusion among the group, the therapist, and one special member. When not properly addressed, this situation can contribute to the stagnation and possible dissolution of the group. Thus, the PGM provides a key to understanding the perils of unrecognized emotional undercurrents and their consequences. PMID- 9217348 TI - An analytic group for sexually abused men. PMID- 9217347 TI - Enhancing Latin American women's self-concept: a group intervention. AB - This study attempts to explore the self-concept of Latin American immigrant adult women and measure the effectiveness of an "Enhancing Self-Concept" workshop on improving their self-concept. The sample consisted of 74 adult women aged 20 to 60 who were selected from first-level second-language courses from various educational institutions in Montreal. Participants were randomly assigned to either an experimental group who received an "Enhancing Self-Concept" workshop or a control group who received a series of information sessions. Outcome measures consisted of scores on the Tennessee Self-Concept Scale (TSCS) and a Post Intervention Questionnaire (PIQ). Participants were tested before and after their respective treatments, depending on the measure used. The results are discussed in relation to effectiveness of the "Enhancing Self-Concept" workshop in improving immigrant Latin American women's self-concept. Implications and suggestions for future research are included. PMID- 9217349 TI - The risk of relapse after multidrug therapy in leprosy. PMID- 9217350 TI - Detection of S-100 protein and anticeramide antibodies in leprosy patients by ELISA. AB - The status of assay for S-100 antigen protein and anticeramide antibodies in serum in understanding nerve damage in different forms of leprosy were evaluated by the enzyme immunoassay. Based on the clinical and smear examination, patients were classified as indeterminate (Ind), tuberculoid (TT), borderline tuberculoid (BT), borderline lepromatous (BL) and lepromatous (LL). Antibody levels against ceramide were observed in sera of leprosy patients with 37.5% of Ind, 28% of TT, 66% BT, 78% BL and 62% LL patients positive as against 8% endemic normal sera. The mean OD ranged from 0.141 to 0.275 in different groups of leprosy. In contrast, S-100 was detected in 71.4% Ind, 88.8% TT, 76.4% BT, 100% BL and 95.8% LL, while 5% of ENL samples were positive for S-100 antigen. Mean S-100 levels in these different categories of patients were significantly higher Ind--0.45 ng/ml, TT--0.32 ng/ml, BT--0.23 ng/ml, BL--0.23 ng/ml, LL--0.19 ng/ml as compared to that of normal 0.07 ng/ml. In general S-100 seems to be a more sensitive and reliable marker than anticeramide antibodies for nerve damage. Five out of 7 indeterminate cases show increased levels of S-100, showing an extent of nerve damage similar to that of TT and could be a useful marker for assessing nerve damage in indeterminate patients for better management. PMID- 9217351 TI - Lymphostimulatory and delayed-type hypersensitivity responses to a candidate leprosy vaccine strain: Mycobacterium habana. AB - Lymphostimulatory and delayed-type hypersensitivity (DTH) immune responses to a candidate antileprosy vaccine Mycobacterium habana have been quantified in inbred AKR mice. M. habana vaccine in three physical states, live, heat-killed and gamma irradiated, was given intradermally to separate groups of mice and after 28 days these mice were given subcutaneous challenge with heat-killed M. leprae and heat killed M. habana in the left hind footpad. Live BCG vaccine alone and in combination with gamma-irradiated M. habana were also compared similarly. A sufficient degree of DTH response was generated in mice by M. habana vaccine in all physical forms against two challenge antigens (lepromin and habanin). The BCG combination with M. habana did not increase the DTH response indicating internal adjuvanticity endowed in M. habana. The active hypersensitivity of immunized mice was transferable to syngeneic mice by the transfer of sensitized cells from the donor to the recipient mice intravenously. M. leprae-infected Rhesus monkey PBMC have shown comparable stimulatory response with M. habana (sonicate), and M. leprae (sonicate) antigens. The possibility of developing M. habana as a candidate antileprosy vaccine is discussed. PMID- 9217352 TI - Higher incidence of viable Mycobacterium leprae within the nerve as compared to skin among multibacillary leprosy patients released from multidrug therapy. AB - As identified by a significant growth in the footpads of immunosuppressed mice, the incidence of viable bacteria in a group of 26 multibacillary (BL-LL) patients released from multidrug (MDT) treatment was found to be two times more in the nerves (46%) as compared to skin (23%). Evidently there was a positive correlation between the overall bacterial load and the incidence of viable organisms. Bacterial growth was also observed in two out of five cases where neither the skin nor the nerve homogenate had shown any presence of acid-fast bacilli. Histopathology of biopsies, skin as well as nerve, including those having viable bacteria did not show any features of active disease. PMID- 9217353 TI - Tuberculosis control is good for established leprosy programmes. AB - Tuberculosis (TB) control was introduced into part of the Danish Bangladesh Leprosy Mission's large leprosy control programme in 1994. This was in line with the Government's policy of combining leprosy and TB control. We report our experience with integration. Leprosy case-finding has increased during the period, and staff satisfaction and morale has also risen despite the larger workload. We observed that the field work skills of leprosy workers was brought to bear in a very positive way on TB control. TB patients suffer considerable impoverishment as a result of their illness, paralleling the social dehabilitation often seen in leprosy sufferers. TB control is good for established leprosy programmes. PMID- 9217354 TI - Surgery for neuritis in leprosy: indications for and results of different types of procedures. AB - From December 1988 to December 1992, 129 surgical procedures were performed on the peripheral nerves of 64 leprosy patients at the Hospital Cardinal Leger de l'Institut Fame Pereo for leprosy control in Haiti. Sixty-four patients totalizing 129 nerves with sufficient clinical data form the basis of this study. Based on the retrospective analysis of the operated cases, a new classification built on macroscopic findings of the involved nerves is presented. Five grades, according to the presenting aspects of these nerves, are set up as guides for different surgical procedures to be performed on the nerves: external decompression for the lesser grades I and II, intraneural neurolysis, interfascicular neurolysis for the higher grades III and IV, cleaning, and debridement for grade V. The final results are discussed. This new macroscopic grading done at surgery helps to minimize the aggressive procedures performed on nerve trunks, decrease the morbidity of surgical action on the nerve vascular structures, and consequently, preserves all possible sensory and motor functions of a nerve. PMID- 9217355 TI - An investigation of attitudes, beliefs and behaviour of leprosy patients, family members and PHC workers towards multidrug therapy in Yangzhou and Dongtai Districts of China. AB - To improve the operational efficiency of multidrug therapy (MDT) implementation in rural areas, an investigation into the attitudes, beliefs and behaviour of leprosy patients and their family members as well as primary health care (PHC) workers towards MDT was carried out in Yangzhou and Dongtai Districts of China. A sample of 370 leprosy patients, 594 family members and 730 PHC workers was interviewed or investigated individually using questionnaires. The results showed that: 1, the presently used MDT is acceptable to a wide range of patients although a small number of patients have various problems in their treatment; 2, the patients' habit in daily drug administration, their awareness of the risk of default and confidence in MDT have a positive influence in increasing drug compliance; and 3, the supervision and encouragement of family members to patients' treatment which is associated with their knowledge on MDT is also beneficial to patients' drug compliance. However, only half of the PHC workers had a basic knowledge of MDT and a desire to participate in MDT implementation, a finding which clearly calls for urgent attention and improvement. In order to ensure the effective implementation of MDT, there is a need to educate leprosy patients and their family members as well as PHC workers to establish the patients' correct awareness of MDT, obtain the family support and motivate the PHC workers. PMID- 9217356 TI - Relapse in a borderline-tuberculoid case of leprosy 5 years after the release from rifampicin monotherapy. PMID- 9217357 TI - Transepidermal elimination of lepromatous granuloma: a mechanism for mass transport of viable bacilli. AB - A 35-year-old male with lepromatous leprosy showed significant progression of the disease on initial examination. Along with typical lepromatous skin lesions, many scar-forming lesions were present, mainly on his extremities. Some lesions showed erosive surfaces. From clinicopathological findings, these lesions were suspected to be due to the partial excretion of intradermal lepromatous granulomata by 'transepidermal elimination'. Increased local volume, which might be due mainly to rapidly growing lepromatous infiltration before chemotherapy, is suspected of triggering this phenomenon. There is no doubt that many fresh Mycobacterium leprae were included in these excretions. After the initiation of chemotherapy, no new scar-forming lesions were observed. PMID- 9217358 TI - Ulnar abscess: 4 months after release from control with paucibacillary-multidrug therapy. PMID- 9217359 TI - Need for intensive leprosy case finding for the elimination of leprosy. PMID- 9217360 TI - An assessment of performance 'contract leprosy workers' in a National Leprosy Eradication Programme. PMID- 9217361 TI - Individual goals and training in leprosy: need for revision of current strategy. PMID- 9217362 TI - [Gastric ulcer and cancer]. PMID- 9217363 TI - [Combined effect of adoptive immunotherapy (AIT) with lymphokine-activated killer (LAK) cells and interleukin-2 (IL-2) and chemotherapy in tumor-bearing mice]. AB - We investigated beneficial effects of AIT with anticancer agents on survival of subcutaneous tumor-bearing mice and suppression of artificial lung metastasis, and optimal schedule of administration of each treatment in vivo. 7-8 weeks old C 57 BL/6 mice were inoculated s.c. with 5 x 10(6) B 16 melanoma cells, or i.v. with 2 x 10(5) B 16-F 10 melanoma cells. Mouse splenocytes were cultured with 3.5 x 10(3) JRU/ml interleukin 2 for 14 days, and induced LAK cells were harvested. Anticancer agents (Cx), CDDP or MMC were given i.p. in mice. 1 x 10(7) or 5 x 10(7) LAK cells were given either s.c. around the tumor or i.v. respectively, and 1.4 x 10(5) JRU/mouse IL-2 was administered s.c. for 6 days after LAK cell injection. Therapy groups were as follows #1: Cx day 3, AIT day 3-8. #2: Cx day 3, AIT day 6-11, #3: Cx day 8, AIT day 3-8. In therapy groups #1 and #2, we observed additive effects of AIT and anti-cancer agents in life-prolongation and suppression of lung metastasis. It was also shown that LAK induction in vivo was augmented by anticancer agents in groups #1 and #2, which might represent one of the mechanisms behind observed additive effects. Furthermore, our results suggest that therapeutic effects of the combination of AIT and anticancer agents depend on the schedule of administration. PMID- 9217364 TI - [Inhibitory effects of portal venous injection of type II collagen in collagen induced arthritis]. AB - Collagen-induced arthritis (CIA) is useful animal model for human rheumatoid arthritis. We investigated the inhibitory effects of portal venous (p.v.) injection of type II collagen (CII) in CIA. The arthritis was suppressed by p.v. injection of CII before immunization for CIA induction. The p.v. route was more effective than intravenous or intragastric routes in the induction of tolerance in CIA. The dose of CII necessary for CIA suppression was 10 micrograms/20 g body weight in p.v. injection. Both anti-CII IgG and anti-CII IgG 2 a levels in serum were reduced in mice injected CII before induction of CIA. However, anti-CII IgG 1 levels did not differ between mice injected with CII and mice injected with buffer alone. Thus, the specific reduction in anti-CII IgG 2 a levels in mice treated by p.v. injection before immunization suggests that the suppression of CIA could be responsible for hypofunction of Th 1 cells. Reduction of anti-CII IgG and suppression of arthritis were observed when CII was injected through portal vein after immunization for CIA as well. PMID- 9217365 TI - [Induction of passive cutaneous anaphylaxis by oral administration of antigen]. AB - Passive cutaneous anaphylaxis (PCA) reaction was developed by Ovary et al. as an animal model of mainly human type I allergic inflammation reaction. This is the most sensitive reaction for the detection of cutaneously sensitizing antibodies and provides a very effective means by which to investigate immunological reactions concerning the mechanisms of development and inhibition of allergic reactions, levels and specificity of antibodies, and the structure of antigens. These cutaneous inflammations mimic atopic dermatitis. Food ingestion has been pointed out as one of the worsened factors of atopic dermatitis. However, the body is generally protected against the invasion of high molecular substances such as non-ingested food by several barriers including digestive enzymes that break down complex food molecules into simpler substances and gastrointestinal mucosae. Accordingly, oral ingestion of food antigen seems to be physiologically and immunologically in conflict with the occurrence of dermatitis. With a view to determining whether allergic dermatitis occurs after oral ingestion of food, the present study was carried out on animals, utilizing the PCA reaction. C 57 BL/6 Ncr j mice were immunized with immunogen derived from commercially obtained eggs. Wistar rats were used as a model of PCA reaction. The results of the present investigation are summarized as follows. 1) Blue spots of 10 mm diameter were observed as a PCA reaction 50 min or more after oral administration of antigen, and the blue spots reached maximum size (20-21 mm, 1.8-1.7 micrograms) after 90 120 min. The PCA reaction was induced 20 min or more after intravenous administration of antigen. When the maximum reaction was compared between the oral and the intravenous routes after 50 min (29.5 micrograms) and 90 min (1.8 micrograms) respectively, there was about a 16-fold difference in the capacity to induce inflammations. 2) The maximum PCA reaction values were 100 and 1,600 for the oral and intravenous routes, respectively, there being a 16-fold difference between the two routes. 3) The minimum antigen concentration required to induce the PCA reaction was 10 mg/ ml for the oral route and 0.01 mg/ml for the intravenous, there being a 1,000-fold difference between the two routes. 4) Reactions with anti-egg mixture antibody and main egg constituents were specific. The PCA inhibition test results confirmed that the undigested structural portion of antigen was associated with the induction of PCA. The present investigation demonstrated that there existed a mechanism by which type I allergy is induced via the gastro-intestinal tract. This fact indicates that part of the food ingested undergoes no change in its molecular structure when transferred to the blood, thus acting as a PCA inducing antigen. This phenomenon suggests that this animal model of human type I allergic dermatitis is a useful system that strongly suggests the association of food antigen with the development of allergic dermatitis. PMID- 9217366 TI - [Allergic contact dermatitis due to eye drops. Their clinical features and the patch test results]. AB - We studied the clinical manifestations and causative agents in cases with allergic contact dermatitis due to eye drops who consulted the Department of Dermatology, Nippon Medical School during the period of 9 years between January, 1987 and December, 1995. Among 66,165 cases who visited the department during the studied period, 3,903 were suspected as contact dermatitis or drug eruption and underwent patch test. 141 of 3,903 cases (3.6%) were patch tested with eye drops and 49 cases (34.8%) reacted positively and were diagnosed as allergic contact dermatitis. Allergic ingredients were tested in 36 cases by the patch test with every ingredient. The 49 cases diagnosed as allergic contact dermatitis by eye drops, included 17 males and 32 females. 81.6% of the patients were aged over 40 years. The most frequent ages were between 60 and 69 years. The period from onset to the first visit to us was less than two weeks in 54.1% of the cases, but 3 cases consulted us after more than six months. The determined allergens are noted in order of frequency as follows: 1) fradiomycin sulfate in 14 cases, 2) ketotifen fumarate in 6 cases, 3) dibekacin sulfate in 3 cases, 4) befunolol hydrochloride in 3 cases, 5) phenylephrine hydrochloride in 3 cases and 6) benzalkonium chloride, the preservative of eye drops in 3 cases. Fradiomycin sulfate showed cross reactions to other aminoglycosides in the present cases. Every ingredient should be tested as much as possible, because there is a possibility that a preservative or a vehicle ingredient may be the allergen, though the incidence is low. PMID- 9217367 TI - The effect of dacarbazine and vincristine sulfate on human melanoma cell lines. In vitro analysis of interaction on DNA synthesis, RNA synthesis and protein synthesis. AB - The effect of anticancer agents, dacarbazine [DTIC (dimethyl-triazeno-imidazole carboxamide)] and vincristine sulfate (VCR) on two human melanoma cell lines (G 361 and MeWo) was investigated. First we estimated the growth curve of two cell lines by using an isotope labelling technique with DNA, RNA and protein precursors to find the most appropriate conditions, and then, DTIC and VCR were added at various concentrations. When the drug was added alone, DNA, RNA and protein synthesis were suppressed in a dose-dependent fashion and the pattern of responses was similar between the two. When the two agents were administered in combination, responses were variable depending on the combinations of the concentrations of these agents. It was also observed that the response on DNA synthesis is not similar to that on RNA or protein synthesis, and that unbalanced growth can happen when two anticancer agents, whose mechanisms of action against tumour cells, are different, are administered at the same time. These observations also differed between the two cell lines. It suggests that these diverse responses of melanoma cells against chemotherapeutic agents hinder the establishment of a sensitive combination chemotherapeutic regimen. PMID- 9217368 TI - [Whether allergic contact dermatitis is necessary to induce systemic immunization by applied hapten]. AB - Hapten specific lymphocyte proliferative assay was used to measure systemic immunization induced by epicutaneously applied hapten, instead of the assay measured by ear swelling or footpad swelling. DNFB was painted on the shaved back of C 57 BL/6 on two consecutive days and 4 days later. DNFB sensitized lymph node cells (LNC) were obtained from the regional lymph nodes. Spleen cells were pulsed with an antigen after irradiation and used as antigen presenting cells (APC). Various numbers of LNC were cultured with various number of APC to determine the highest lymphocyte proliferation. This was observed when 2 x 10(5) cells/well APC were cultured 4 x 10(5) cells/well LNC. In next step, this assay was used to investigate whether contact dermatitis is a required subject or not to sensitize. FB could not induce contact dermatitis on the painted region not only clinically but also histologically. FB, however, was able to induce hapten specific lymphocyte proliferation. This fact demonstrates that contact dermatitis is not always necessary in sensitization through epicutaneous application of hapten. PMID- 9217370 TI - [Somatosensory evoked potentials (SEPs) as an intraoperative monitor in lower abdominal surgery during lumbar epidural anesthesia]. PMID- 9217369 TI - [The role of hepatic sinusoidal cells in homing of lymphocytes. Preferential hepatic trapping of PNA(+) lymphocyte and its modulation during liver regeneration]. AB - It was demonstrated that the reaction of some cell species to peanut agglutinin (PNA) involved the maturity of the cells. We investigated the selective adhesion of circulating hemopoietic cells and lymphoid cells to hepatic sinusoidal cells. We experimented with thymocyte (TYC) models that could be easily isolated into PNA positive [PNA(+)] and PNA negative [PNA(-)] subsets, and transfused these subsets, labeled with 51Cr into the isogenic mice, then measured the radioactivity in the kidney, lung, spleen, liver, Peyer's patch (P.P.), and thymus, after 48 hrs. The radioactivity (% total dose recovered) in the liver and spleen represented the degree of cellular distribution and indicated that a significantly higher dose had accumulated in the liver and spleen than in other organs. Additionally, TYC-PNA(+), recognized to be immature cells, demonstrated high aggregation in the liver, while TYC-PNA(-), recognized to be mature cells, demonstrated high aggregation in the spleen. Similar results were observed in our subsequent experiment using wheat germ agglutinin (WGA), other indicator of maturity, and spleen cells (SPC). Next, we transfused TYC-PNA(+) to mice at various intervals after 70% hepatectomy. The degree of aggregation (radioactivity/weight) in the liver was reduced 1 day after hepatectomy, and that in the spleen increased proportionately. Whereas the aggregation degree of TYC PNA(-) in the same experiment did not change significantly. We postulated that hepatic sinusoidal cells recognize and, sequentially, trap specific carbohydrates on lymphoid cells by a lectin, PNA or WGA, like receptor. Reviewing our previous report, adhesion to Kupffer cells induced SPC activation and proliferation, this recognition and trapping mechanism seems to play an important role of intrahepatic hematolymphoid system. PMID- 9217371 TI - [Catheter ablation therapy for cardiac arrhythmias]. PMID- 9217372 TI - [Primary malignant lymphoma of the ischium]. PMID- 9217373 TI - On the mechanism of cAMP-dependent modulation of single inward-rectifier K+ channel kinetics in the mammalian heart. PMID- 9217374 TI - [Evaluation of coronary artery bypass grafts with magnetic resonance imaging]. AB - Currently, the efficacy of magnetic resonance imaging (MRI) for evaluating coronary artery disease has been reported. In this study, we have evaluated the usefulness and the problems of MRI for evaluating the patency of coronary artery bypass grafts. Thirty-five patients who received coronary artery bypass grafting (CABG) were evaluated by using MRI for determining the graft patency compared with conventional coronary angiography. There were 30 men and 5 women. The mean age was 61.2 years (range 45 to 75). The 35 patients had a total of 92 grafts (28 internal thoracic artery, 7 gastroeploic artery and 57 saphenous vein grafts). Magnetic resonance coronary angiogram (MRCA) was performed with SIGNA HORIZON 1.5 T (GE Inc.) by using 2D-FASTCARD sequence. All patients underwent imaging in the transverse and coronal planes, most had imaging in the sagittal plane, and a few had in the oblique plane. By using MRCA, 82 of 90 grafts were diagnosed correctly as patent, and 1 of 2 grafts were diagnosed correctly as occluded. Thirty-four of 40 LAD grafts (85%), 20 of 22 RCA grafts (91%) and 29 of 30 Cx grafts (97%) were correctly evaluated. The efficacy of MRCA for evaluating the patency of coronary artery bypass grafts was recognized. But the sternal wire (stainless steel) and hemoclip interfere with the interpretation and reduce the sensitivity. Higher sensitivity may be obtained by changing the material of the sternal wires and hemoclips at coronary surgery. PMID- 9217375 TI - [A clinical study of retrograde cerebral perfusion in long duration]. AB - The safety limit of retrograde cerebral perfusion (RCP) in aortic arch reconstruction was evaluated in comparison with long (> or = 60 min: group A = 28 cases) and short (< 60 min: group B = 88 cases) RCP time (RCPT). RCPT was 60-94 min in group A and 24-59 min in group B. Hospital mortality was 7.1% and 6.8% in group A and B, respectively. There was no case who was died due to cerebral insufficiency. Postoperative neurological complication was observed 3 in group A and 3 in group B. There was no significance between 2 groups. There were no significant differences in the time to awake, ICU stay, electroencephalogram, and cognitive functions between 2 groups. Based on these results, the safety limit of RCP is over than 60 min and, maybe, about 90 min. We monitored the increase of deoxygenated hemoglobin (HbR) during RCP using a near infrared spectroscopy (n = 55). Since HbR increased gradually during RCP, HbR reflects oxygen metabolism non invasively in the brain. HbR can be one of useful indexes of the brain protection during RCP. PMID- 9217376 TI - [Area reduction effects on atrial fibrillation inducibility]. AB - Atrial fibrillation inducibility and sustenance depend upon the atrial electrophysiologies such as wavelength and upon the anatomical factors such as atrial area. To determine the mechanism of prevention of atrial fibrillation by atrial mass reduction, 20 adult mongrel dogs underwent the right atrial isolation procedure and atrial fibrillation duration, functional refractory period, conduction velocity, wavelength, atrial area and weight were examined pre- and postoperatively. The animals were divided into two groups: right atrial isolation group (RAI-group, n = 10) and control group (C-group, n = 10) in which cardiopulmonary bypass was undergone for the identical duration and the right atrial isolation was not performed. Atrial fibrillation duration significantly prolonged from 9.2 +/- 2.5 sec preoperatively to 43.6 +/- 0.4 sec postoperatively in the control group. However, in the RAI group, the duration significantly shortened from 11.4 +/- 2.7 preoperatively to 1.2 +/- 10 sec postoperatively. The wavelength at the non-isolated atrium did not show significant difference between the control group and the RAI group. The atrial area decreased from 61.9 +/- 4.1 cm2 to 46.3 +/- 3.5 cm2 postoperatively in the RAI group, while the area did not change in the control group. We conclude that the reduction in the effective atrial area affects the inducibility of atrial fibrillation. PMID- 9217377 TI - [Effect of aprotinin on bleeding during and graft patency after coronary artery bypass grafting]. AB - The purpose of this study was to evaluate the effect of aprotinin on intraoperative bleeding and graft patency in 200 patients undergoing elective CABG. Patients were classified into three groups; patients receiving 1500000 KIU of aprotinin (group A), 300000 KIU of aprotinin (group B) and no aprotinin as the control (group C). The groups were almost the same as to age, sex, number of grafts and use of ITA. Intraoperative bleeding and reexploration were significantly decreased in group A compared with group B and C. All patients underwent coronary angiography between the 4th and 8th post-operative weeks. Graft patency was significantly decreased in group A (SVG: 90.4%, ITA: 91.1%) compared with group B (SVG: 95.3%, ITA: 100%) and C (SVG: 95.5%, ITA: 99.2%). Perioperative myocardial infarction increased in group A (3.7%). Aprotinin was effective in reducing intraoperative bleeding in CABG, but decreased early graft patency after CABG. It is suggested that aprotinin should not be used routinely in CABG unless a patient has bleeding tendency which is likely to be seen in emergency and redo surgery. PMID- 9217378 TI - [Effects of pre-operative administration of steroids on the serum interleukin (IL)-6, IL-8 and organ injuries in replacement of thoracic aorta]. AB - To investigate whether pre-operative steroids administration decreases the post operative serum IL-6, IL-8 and prevents organ injuries, we prospectively studied patients undergoing elective replacement of thoracic aorta using an extracorporeal circulation. Six of 10 patients (group S) were pretreated with methylprednisolone 500 mg 2 hours before operation while the other 4 patients (group C) were not. Though post-operative serum IL-6, IL-8, C-reactive protein and amylase elevated in group C, the elevations were significantly decreased in group S (p < 0.05). These findings show that in replacement of thoracic aorta, pre-operative steroid administration decreases the post-operative elevations of serum inflammatory cytokines (IL-6 and IL-8) and may prevent organ injuries. PMID- 9217379 TI - [Relapse after video-assisted thoracoscopic surgery for spontaneous pneumothorax]. AB - Postoperative relapse was noted in 8 (14.8%) of 54 patients who underwent video assisted thoracoscopic surgery for spontaneous pneumothorax followed for more than one year. Five of the eight patients with relapse were among the 15 initial stage cases. Concerning treatment, only drainage was performed in 1 patient, and thoracotomy was performed in 5 patients. Findings on additional surgery consisted of missed cystic lesions, in 2 patients, a small opening adjacent to a suture caused by an autosuture device in 1 patient, and a cyst around a suture in patients. There was no missed cystic lesion after the 15th case. In subsequent cases, there were changes at the periphery of a suture. In patients with other diseases, there were no changes observed when an autosuture device was used under thoracoscopic guidance. Therefore, the changes may be associated with hypertonus of the emphysematous lung at the periphery of the site where an autosuture device was used. Thoracoscopic surgery for spontaneous pneumothorax using an autosuture device has limitations such as the presence of a dead angle, insufficient evaluation of emphysematous changes in the collapse lung, a large volume of resected tissue, and hypertonus of the peripheral lung. These limitations may contribute to a high recurrence rate of spontaneous pneumothorax. To reduce the recurrence rate, conditions and problems in cases showing relapse should be clarified before selecting treatment including pleurodesis in individual cases. PMID- 9217380 TI - [Clinical evaluation of heparin concentration and activated clotting time monitoring (HEPCON HMS) system]. AB - The HEPCON HMS system provides both activated clotting time (ACT) and accurate whole blood heparin concentration measurements. We evaluated the impact of heparin and protamine administration using this system on the incidence and treatment of bleeding after performing a cardiopulmonary bypass. Patients were randomly divided into two groups. Heparin and protamine administration during extracorporeal circulation was determined by classical ACT management in Group A (n = 15) and by HEPCON HMS system in Group H (n = 19). There were no statistical differences in the coagulation factor, the postoperative chest tube drainage or the blood products used between the two groups. Patients in Group H received a higher dosage of heparin and a lower dosage of protamine compared with Group A. By facilitating the maintenance of a therapeutic heparin concentration and by determining an appropriate protamine dosage, the HEPCON HMS system may be useful in managing extracorporeal circulation. PMID- 9217381 TI - [Experimental study of the effectiveness of immunosuppression therapy using FK506 and 15-deoxyspergualin in concordant xenotransplantation]. AB - The effect of combination therapy with FK506 (FK) and 15-deoxyspergualin (DSG) was investigated in a concordant xenotransplantation model in which hamster hearts were implanted into LEW rats. Forty xenograft procedures were carried out in this study. Recipients were divided into eight groups. Control Group received no therapy. Group A, B, and C were administered different dosages of FK; 0.75, 1.0 and 1.25 (mg/kg/day), respectively. In Group D and E, DSG at 10 and 20 mg/kg/day was administered after the transplantation. In Group F and G received combined administration of FK and DSG (Group F; FK 0.5 + DSG 10, Group G; FK 0.5 + DSG 5). Recipients were injected with immunosuppressive agent intraperitoneally from day 0 until cessation of heart beat. At the time of grafting, splenectomy was performed in all groups. FK treatment of the recipients did not lead to significant improvement in graft survival, 7.2 +/- 0.8 and 6.3 +/- 0.7 days, compared with 3.5 +/- 1.23 days in Control. DSG groups showed xenograft survival of 11.5 +/- 0.9 and 8.1 +/- 1.6 days, respectively. The combined therapy prolonged graft survival to 33.67 +/- 11.37 days in Group F, and to 47.3 +/- 11.02 days in Group G. Combined therapy produced complete suppression of antibody formation, but in FK 0.5 + DSG 10 mg/kg/ day treated group, there was abrupt elevation of anti-hamster antibody titer at the time of rejection. Only in combined therapy groups did recipients obtain long-term graft survival, but recipients receiving treatment with FK 0.5 + DSG 10 (mg/kg/day) suffered from severe emaciation, losing nearly 31% of their body weight. However, animals that received a low dose of DSG in combined therapy groups did not lose body weight over 8 weeks. Combination therapy with FK and DSG was shown to prolong graft survival time effectively, and FK 0.5 + DSG 5 (mg/kg/day) was an appropriate dose in xenotransplantation with minimal side effects. PMID- 9217382 TI - [Investigation of complications of bronchial artery embolization using superselective catheter system and platinum coil-major complications as vascular detachment]. AB - We have performed bronchial artery embolization (BAE) in patients with hemoptysis using a superselective catheter system and a permanent-emboli coil. We performed an investigation because major complications were observed. Of a total of 57 BAE using this system, 7 cases of major complications as detachment of the large vessel were found. Although BAE using this system is effective because permanent emboli can be selectively in-dwelled in abnormally developing vessels, detachment of vessels were developed in 7 cases. They are found more frequently for embolization of the left bronchial artery and full attention should be paid to this phenomenon when performing this surgery. PMID- 9217383 TI - [Urgent thoracotomy for injuries to the tracheobronchial tree due to blunt trauma -a seven cases report and a literature review of 32 cases in Japan]. AB - Surgery for injuries to the tracheobronchial tree due to blunt trauma occurs over three periods: an emergency period requiring emergency room thoracotomy, an urgent period, and a late period. We observed seven cases in which thoracotomy (tracheobronchial plasty and pneumonectomy/lobectomy) was carried out during the urgent period. Thirty-two similar cases reported in Japan possessed features with the same patterns. It has been considered that lesions were frequently injured near the carina and that the bronchus would be injured symmetrically. However, both our experience and a literature review shows that only 59% of the cases (23/39) were injured near the carina and that 88% of bronchial injuries (14/16) occurred on the right side. This report also reviews the following clinical symptoms and roentogenological signs that contributed to the diagnosis: 1) dyspnea or respiratory distress, 2) subcutaneous and/or mediastinal emphysema, 3) fallen lung sign, and 4) upper rib fractures. Although a flexible bronchofiberscope was performed only in 24 cases (75%) out of the 32 reviewed patients and only 20 (83%) out of the 24 was diagnosed, the examination should be performed for all the patients even with minor or possible injury of the tracheobronchial tree. The results also suggested that an immediate and accurate diagnosis and subsequent emergency thoracotomy may improve the prognosis of the injury for patients during emergency period. PMID- 9217384 TI - [Serratia marcescens prosthetic mitral valve endocarditis associated with hemolytic anemia]. AB - A 57-year-old female who had been performed mitral valve replacement (MVR) using 31 mm prosthetic valve 32 months before entered the hospital for the evaluation of long standing severe hemolytic anemia without infectious sign. Transesophageal echocardiogram revealed a moderate sized vegetation on the atrial site of the prosthetic valve. The size and number of the vegetation were increased after deterioration of infectious illness. Blood culture grew serratia marcessans and alpha-hemolytic Streptococcus. Re-MVR was carried out with the diagnosis of prosthetic valve endocarditis (PVE). As the symptom of PVE, hemolytic anemia without infectious sign is a rare condition. TEE is an useful method to make diagnosis of PVE by detecting the vegetations and evaluating their change of size and methods and to evaluate the effectiveness of the treatment. PMID- 9217385 TI - [A case of ruptured aortic arch aneurysm with hemorrhagic cardiac tamponade]. AB - The ruptured thoracic aortic aneurysm is still a dramatic even with very poor outcome, whereby its survival depends largely on early diagnosis and operation. We report a successful case of aortic arch replacement for ruptured aortic arch aneurysm with cardiac tamponade. Lethal hemopericardium causing cardiac tamponade is most commonly seen as a complication of acute myocardial infarction or acute aortic dissection, and subsequent rupture of the heart or ascending aorta leads to the rapid accumulation of blood within the poorly distensible pericardial sac. Our case was operated upon emergency basis due to hemopericardium. On operative findings, the aortic aneurysm located the minor curvature of aortic arch and was a huge saccular shape. In surgical procedure, the total arch replacement was completed using selective cerebral antegrade perfusion with deep hypothermia. Postoperative course was uneventful and no cerebral complication was observed after surgery. PMID- 9217386 TI - [A surgical case of total anterior papillary muscle rupture after acute myocardial infarction]. AB - Mitral regurgitation (MR) due to rupture of the papillary muscle is one of the most serious complications of acute myocardial infarction (AMI) as well as ventricular septal perforation and ventricular free wall rupture. We experienced a case of complete rupture of the anterior papillary muscle. A 68-year-old man experienced an episode of dyspnea. Electrocardiographic findings were consistent with postero-lateral infarction. Massive MR is present on color Doppler imaging. He was transferred to our hospital for urgent operative indication because of papillary muscle rupture due to AMI. Six hours after the onset, the operation was performed with intra-aortic balloon pump in place. The anterior papillary muscle was ruptured completely in the mid portion. He underwent a mitral valve replacement with a SJM prosthetic valve. There is a few cases with successful urgent surgery for a complete rupture of anterior papillary muscle. PMID- 9217387 TI - [Urgent coronary reoperation under the beating heart via the left thoracotomy]. AB - An 80-year-old female, who had received coronary artery bypass grafting (CABG) 12 years ago was admitted to our hospital because of unstable angina. The vein grafts to the left anterior descending artery (LAD) and circumflex artery (Cx) were both occluded. Chest computerized tomography showed a severely calcified aorta. An intra-aortic balloon pump was inserted and an urgent reoperation was performed. The patient was positioned for an anterolateral left thoracotomy. The chest was entered through the forth intercostal space. The pericardium was opened parallel to the phrenic nerve. Femorofemoral bypass was instituted. Anastomosis of the saphenous vein graft and the diagonal branch and anastomosis of a second saphenous vein to the old SVG just proximal to the anastomotic site of Cx were performed under a beating heart. Proximal anastomosis of the two saphenous veins and the left subclavian artery was then performed. The patient had an uncomplicated postoperative recovery. A postoperative angiogram showed that the new SVG anastomosed to the diagonal branch was patent while the other new SVG anastomosed to the old SVG/Cx was occluded. The patient was discharged and is now free from angina. PMID- 9217388 TI - [A case of total arch replacement for traumatic aortic arch rupture]. AB - We report a case of aortic arch rupture due to blunt chest trauma. A 64-year-old woman was crushed under a wood when she was working. On her arrival to our hospital, she was in shock state due to cardiac tamponade. Her chest roentogenogram showed fracture of her right-sided ribs and mediastinal widening. Her chest CT revealed hematoma around the aortic arch. Her transesophageal echography (TEE) demonstrated an intimal flap in the aortic arch. An emergency operation was performed immediately under the hypothermic circulatory arrest with a selective cerebral perfusion. As the intimal disruption was found to be extended to three fourths of the circumference of aortic arch, total arch replacement was performed. In this case, the TEE was useful for the immediate diagnosis of the aortic rupture. This was the first successful case of total arch replacement for a traumatic arch rupture. PMID- 9217389 TI - [Giant localized fibrous pleural mesothelioma accompanying lung metastasis--a case report]. AB - We experienced a case of giant localized fibrous pleural mesothelioma accompanying lung metastasis. The patient was a 26-year-old woman, who was pointed out elevation of the left diaphragma by the chest roentgenogram in 1992 but left it as it was. In 1995, a giant tumor occupying 1/2 or more of the left thoracic cavity was detected by chest CT and MRI. A giant localized pleural mesothelioma was suspected since there was no accumulation in Ga scintigram and an operation was performed. The tumor originated without pedicle and with invasion of the lung from the pleura of the left S8 and it was excised by separating the left lower lobe at 7-cm distance from the tumor. The tumor was 20 x 12 x 11 cm in size weighing 1700 g and the histopathological diagnosis was localized fibrous pleural mesothelioma. A lung metastasis of 1.0 cm in size was found in a part of the left lower lobe resected at the same time. The postoperative course has been satisfactory and there is no sign of recurrence at present. There are occasional cases of recurrence associated with a localized fibrous pleural mesothelioma even though it is judged benign. In the present case, a metastatic focus was already present at the time of initial surgery and is considered a valuable case. PMID- 9217390 TI - [Successful surgical treatment of double-outlet right ventricle with intramural coronary artery, using the modified Aubert procedure--a case report]. AB - The modified Aubert procedure and left ventricular outflow tract reconstruction were performed successfully for double-outleft right ventricle with subpulmonary ventricular septal defect, which showed an unusual Shaher type 5a coronary artery pattern. This pattern was characterized by two coronary ostia arising from the right septal sinus and intramural segment in a left trunk of the coronary artery. In this case, the neo-pulmonary artery was reconstructed without prosthetic materials. However, postoperative echocardiography showed no evident supravalvar pulmonary stenosis. The modified Aubert procedure without prosthetic marerids can be used for double-outlet right ventricle and transposition of the great arteries which show dilated prelimonary artery. PMID- 9217391 TI - [Repair of D-transposition of the great arteries associated with double aortic arch]. AB - We report a case of D-transposition of the great arteries associated with double aortic arch. An aortic root angiogram revealed that the left-sided aortic arch was anterior and smaller than the right one and the descending thoracic aorta located on the left side of the spine. Balloon atrial septostomy was performed at one day of age. The patient underwent a simultaneous arterial switch procedure and division of the vascular ring at the isthmus of the left aortic arch through a median sternotomy incision at the age of 16 days. There was persistent postoperative difficulty in weaning the patient from the ventilator. Magnetic resonance images showed re-formation of pseudovascular ring by the connective tissue grown around the divided arch. At the age of 23 days, resection of the remnant of left aortic arch including the left subclavian artery and the diverticulum concomitant with vascular suspension procedure was performed through a left lateral thoracotomy. The patient was subsequently extubated without difficulty and was discharged from the hospital. It is though that a vascular suspension procedure and resection of the subclavian artery are necessary to avoid respiratory obstruction when the great arteries are in an anteroposterior position and Lecompte procedure is performed. PMID- 9217392 TI - [Successful open commissurotomy and resection of dysplastic myxomatous tissue on the leaflet edges in a neonate and an early infant with critical aortic stenosis]. AB - A neonate and an early infant with critical aortic stenosis successfully underwent open commissurotomy and resection of dysplastic myxomatous tissue on the leaflet edges using cardiopulmonary bypass. Case 1: A 31-day-old boy admitted to our unit with shock. Echocardiography demonstrated critical aortic stenosis and severe left ventricular dysfunction (EF = 15%). Case 2: A 12-day-old boy suddenly deteriorated and required resuscitation with ventilation and inotropic support. Emergency operation was required in both cases using cardiopulmonary bypass with systemic hypothermia (30 degrees C). In both cases, the aortic valve was bicuspid and dysplastic with gelatinous myxomatous tissue on the leaflet edges. Commissurotomy and resection of myxomatous tissue were performed. Their postoperation courses were uneventful and they have been free from medication at present. These results suggest that aortic commissurotomy and resection of myxomatous tissue under direct vision may be preferable for critical aortic stenosis with dysplastic aortic valve. PMID- 9217393 TI - [Mitral valvuloplasty for the repair of mitral valve prolapse due to tendon rupture of the posterior leaflet in a father and son--case reports]. AB - The use of mitral valvuloplasty for the repair of mitral valve prolapse in members of a family has been reported only rarely. Patient 1 was a 71-year-old man, and patient 2 the son of patient 1 was a 47-year-old man. Both patients had a grade 3/4 mitral valve regurgitation due to tendon rupture of the posterior leaflet. The procedure was quadrangular resection of the posterior leaflet with annulorrhaphy and ring annuloplasty. The results were good in both cases. Histopathological analysis of a specimen resected from the mitral valve revealed myxomatous degeneration in both cases. These case reports suggest that there is a genetic basis for mitral valve prolapse. PMID- 9217394 TI - [A surgical case of vascular ring (Edwards IIIB) with dysphagia evaluated by esophageal scintigraphy]. AB - Severe esophageal compression due to a vascular ring rarely develops after childhood. We report a case of a 35-year-old man with dysphagia associated with a vascular ring. Radionuclide transit studies were performed before and after surgery for quantitative evaluation of deglutition. Preoperative aortogram disclosed a right sided aortic arch and a retroesophageal left subclavian artery arising from a diverticulum of the arch. Because of the severity of his symptoms, he was taken to surgery. He underwent posterolateral thoracotomy through the fourth intercostal space and division of the ligamentum arteriosum. The esophagus was mobilized from the aortic arch, the arch diverticulum, the pulmonary artery, and the trachea. Postoperatively, he experienced immediate resolution of dysphagia and quickly began eating a regular diet. Although a postoperative esophagogram revealed the esophageal compression, esophageal scintigraphy using 185 MBq 99mTc pertechnetate revealed shortening of the transit time of swallowed water by 1.0 second after surgery. Quantitative evaluation of deglutition in the esophagus by scintigraphy was useful for this patient, since he suffered from psychiatric problems. PMID- 9217395 TI - [A successful surgical case report of acute aortic dissection involving entire sinus of Valsalva]. AB - We successfully performed aortic root replacement for acute aortic dissection, Stanford type A involving the entire sinus of Valsalva, associated with acute anterior wall myocardial infarction and aortic valve insufficiency. A 57-year-old man was admitted complaining of chest pain. An emergency operation was performed after a perfusion catheter was inserted to 99% stenotic lesion of the left anterior descending artery (LAD) on the same day. The dissection extended to both ostia of the coronary arteries and disrupted all commissures of the aortic valve, resulting in severe prolapse of the aortic valve leaflets. Aortic root replacement was performed using a valved conduit. The left main coronary artery was reattached to the graft using interposition technique with a 8 mm diameter woven Dacron tube graft. In addition, the LAD and right coronary artery were bypassed using saphenous vein. The postoperative course was uneventful and the patient was discharged from hospital on the 35th postoperative day. Retaining no aortic sinus and adequate coronary artery reconstruction is important for surgical repair of aortic dissection involving the entire sinus portion of the ascending aorta. PMID- 9217396 TI - [A surgical case of aortic valve stenosis associated with systemic lupus erythematosus]. AB - A 46-year-old woman was hospitalized for aortic valve stenosis (AS) associated with systemic lupus erythematosus (SLE) on April 12, 1996. She had a syncopal episode six months before admission. She was found to have thrombocytopenia, and was diagnosed with SLE by further examination. Irregular genital bleeding was also seen on admission while her SLE was being controlled with steroid therapy. Aortic valve replacement was performed after the steroids had been reduced to avoid excessive bleeding. The aortic valve was the bicuspid with raphe. There was much calcification on the cusps and the annulus, but there were no degenerative changes. The postoperative course was uneventful, and her SLE has been in remission two years after the operation. The management of steroid therapy for SLE patients complicated with cardiovascular disease is discussed. PMID- 9217397 TI - [Ventricular septal perforation due to coronary artery spasm--a case report]. AB - A 70-year-old female was referred due to chest pain and cardiogenic shock. Echocardiogram showed ventricular septal perforation (VSP) with large left to right shunt. ECG indicated ischemia on the left anterior descending region. Instantaneous coronary angiogram was done under the support of IABP. Neither obstructive nor stenotic lesions were found in the coronary arteries. Coronary artery spasm was a likely cause of VSP. The patient underwent emergency surgery within 12 hours from the onset. The VSP, 2 cm in diameter, was located on the postero-apical portion of the ventricular septum. The defect was closed by endocardial patch method using an equine pericardium. Postoperative course was uneventful, and the patient was transferred 3 weeks after the operation. PMID- 9217398 TI - [Stent graft treatment for two cases of DeBakey IIIb dissecting aortic aneurysm]. AB - Two patients with DeBakey IIIb dissecting aortic aneurysms were treated with transluminally placed endovascular stent grafts. Surgery was required for both patients because the false lumens were not thrombosed for several months. Stent graft devices composed of several units of self-expandable Z stents covered with ultra-thin woven Dacron were inserted through 18 Fr sheathes via femoral arteries. The stent grafts were deployed successfully and blood flow into the false lumens was reduced immediately and finally thrombosed without blood leakage from the entries. The endoluminal stent graft treatment is minimally invasive operation in comparison with former surgical operations, and is useful for aortic aneurysms especially in high risk patients. However, improvement of the stent graft devices, including the delivery systems such as the dilator, sheath and pushing rod, which are incomplete, and developing better devices, is required to reduce delivery failure and to make the stent graft treatment more reliable. PMID- 9217399 TI - Clinical ethics. An important role for the consultation-liaison psychiatrist. PMID- 9217400 TI - Consultation-liaison psychiatry and clinical ethics. Historical parallels and diversions. AB - As clinical ethicists increasingly populate hospital settings, a definition of their roles and responsibilities vis a vis those of consultation-liaison (C-L) psychiatrists remains a matter of both interest and uncertainty. Both fields share certain evolutionary and ideological features, yet until very recently, psychiatry has ignored medical ethics, leaving the field to other medical specialties. This estrangement can be explained by psychiatry's traditional suspicion and devaluation of moral philosophy and its more recent wish to be identified more with biomedicine than with the "softer" social sciences and humanities. C-L psychiatry has both a lot to offer and a lot to learn from clinical ethics. PMID- 9217401 TI - Psychiatry and bioethics. An exploration of the relationship. AB - Psychiatrists have been extensively involved in ethics in the general hospital over the past two decades and have functioned in that area in a variety of roles. The basis for psychiatry's strong interest in bioethics can be understood as related to three factors: familiarity with many of the clinical problems that lead to bioethics consultation, the frequent importance of psychiatric aspects of ethics, and the observation that psychiatrists already possess many of the clinical skills necessary for doing ethics work. The particular value of training psychiatrists to serve as ethics consultants, in addition to the importance of their continuing role on hospital ethics committees, is discussed. PMID- 9217402 TI - Consultation-liaison psychiatry and clinical ethics. Representative cases. AB - The skills of the consultation-liaison psychiatrist are enormously valuable in the emerging field of clinical ethics consultation. Expertise in evaluating decision-making capacity is crucial, as is the larger issue of addressing the role that emotional factors play in making life or death decisions. Three cases are reviewed that illustrate the way in which the psychiatric perspective enhances the process of clinical ethics consultation. PMID- 9217403 TI - Making a situational diagnosis. Psychiatrists at the interface of psychiatry and ethics in the consultation-liaison setting. AB - Psychiatrists bring a unique understanding to clinical ethics, but psychiatrists need a precise awareness of the difference between exercising their clinical expertise and facilitating ethical decisionmaking. The author outlines a schema for recognizing and honoring that distinction and illustrates "pseudoethics," "pseudopsychiatry," and "psychiatry/ethics" consultations. The author describes how to make a "situational diagnosis" that includes patient/family issues, staff issues, joint issues, legal/regulatory issues, and ethical issues, thus enabling the psychiatrist to institute an appropriate "hierarchy of interventions": educational, psychological, and ethical. The literature on ethics education for psychiatric practitioners is reviewed and a program is suggested. PMID- 9217404 TI - Neuroticism and extraversion as predictors of health outcomes in depressed primary care patients. AB - Depressed primary care patients (N = 217) were assessed to determine if certain personality characteristics predict health domains independent of chronic disease, demographics, depression, and psychiatric diagnoses. Eleven health variables were used to create three outcome factor scores: disability (e.g., days missed work); somatization (e.g., medically unexplained symptoms); and subjective pain (severity, interference). Neuroticism explained significant variance in all health outcomes independent of the other predictors. Depression and neuroticism interacted in the disability and pain models. Depression was related to health in neurotic patients, while in the absence of neuroticism, little relation between depression and health was observed. Neuroticism may explain why persons with similar health problems have differing levels of disability, pain, and somatization. PMID- 9217405 TI - What, why, and how of consultation-liaison psychiatry. An analysis of the consultation process in the 1990s at five urban teaching hospitals. AB - There is controversy about the role and function of a consultation-liaison (C-L) psychiatrist, as reflected in the ongoing debate about what to call ourselves. To clarify the essential elements of our function, the authors analyzed the process and content of the entire consultation experience from the time of initial consultation to the time of discharge in 50 patients across 5 urban teaching hospitals. The common components of the C-L process, in this pilot study, were identified to be facilitative, consensus-seeking, and interpretative. Implications of these findings for the C-L psychiatrist's role in the general hospital are discussed. PMID- 9217406 TI - Cognitive behavior therapy for functional somatic complaints. The example of chronic fatigue syndrome. AB - Somatic complaints such as pain and fatigue that are unexplained by conventional disease are common in medical practice and are referred to as functional, somatoform, or somatization symptoms. Despite frequent chronicity, disability, and high associated medical costs, patients with these complaints are rarely offered either constructive explanations or effective treatment. In this perspective, a cognitive-behavioral approach to the problem is described, using chronic fatigue syndrome as an example. It is concluded that the utility of the cognitive-behavioral theory and the proven effectiveness cognitive behavior therapy provide the basis for a new evidence-based approach to psychosomatics. PMID- 9217407 TI - Organic mental disorders in the consultation-liaison psychiatry setting. A multi site study. AB - Interventions recommended by consultation-liaison psychiatrists for inpatients they diagnosed as having DSM-III-R organic mental disorder (OMD) were studied to see to what extent specific variables distinguished the OMD patients and differentiated the subgroups of patients with OMD. Prospective data and Mini Mental State Exam (MMSE) scores on 625 consecutive referrals at 3 general hospitals in Australia and the United States were collected by using the MICRO CARES database system. The OMD group differed from the other patients because they were significantly more likely to have been referred for "organic brain syndrome" or "agitation," had less mood disorder and lower MMSE scores, and received more recommendations for antipsychotics and for ward-environment manipulation and fewer recommendations for psychological management. The many differences among the OMD subgroups were also consistent with their DSM constructs. A pilot exploration of the validity of the DSM-IV constructs of cognitive disorder and its subgroups performed on the redistributed data suggested that these constructs have similar usefulness. PMID- 9217409 TI - Recurrent obsessive-compulsive disorder associated with pregnancy and childbirth. PMID- 9217408 TI - Preburn psychiatric history affects posttrauma morbidity. AB - A sample of inpatient, burn-injured adults (N = 95) were assessed upon discharge, and 4 and 12 months later with a structured interview and DSM-III-R criteria. The prevalence of disorder in this sample was contrasted with published data on a representative national community-dwelling comparison group in the National Comorbidity Study. The prevalence of lifetime affective, alcohol, and substance use disorders was significantly higher, and lifetime anxiety disorders significantly lower, in the burn-injured sample. The 12-month postburn prevalences of alcohol, and substance use disorders were significantly greater in the burn-injured sample. The risk of postburn disorder was significantly greater for the subjects who had a preburn history of affective, alcohol, or substance use disorder. The risk for developing posttraumatic stress disorder (PTSD) was elevated in the subjects with a preburn affective disorder but not preburn anxiety disorder. Finally, postburn PTSD was associated with a greater length of stay, and greater preburn comorbidity predicted preburn employment status and tended to lengthen hospitalization. PMID- 9217410 TI - Buspirone in the management of disruptive behaviors due to Huntington's disease and other neurological disorders. PMID- 9217411 TI - Psychiatric manifestations of pituitary tumors. PMID- 9217412 TI - CADASIL presenting as bipolar disorder. PMID- 9217413 TI - Aggressive lithium treatment for mania in a hemodialysis patient with end-stage renal disease. PMID- 9217414 TI - A nifedipine-induced inhibition of lithium clearance. PMID- 9217415 TI - Short gut syndrome, tricyclic antidepressants, and prolonged QT interval syndromes. PMID- 9217416 TI - [Trigeminal neuralgia caused by tortuous vertebrobasilar system--the clinical and imaging features]. AB - Ten (6.8%) out of 146 patients with trigeminal neuralgia (TN) who underwent SPGR MRI and 3D-TOF-MRA from August 1993 to October 1996, were found to have vascular compression caused by a tortuous vertebrobasilar system (TVBS). They were mostly males, demonstrated left-sided predominance, and had ipsilateral hemifacial spasm, compared with other 52 patients whose offending arteries were either superior cerebellar artery (SCA), anterior inferior cerebellar artery (AICA)or posterior inferior cerebellar artery (PICA). The patients who showed vascular compression by TVBS, presented an artery which compresses and dislocates the rootentry zone (REZ) of the trigeminal nerve, presses the brain stem at REZ and simultaneously compresses the REZ of the facial nerve. In addition, the diameters of the two branches of vertebrobasilar artery were not equal. These features indicate that the atherosclerotic change of the offending artery in TN caused by TVBS is more severe than that caused by SCA, AICA or PICA. This change causes an irregular running of artery which leads a strong compression of the trigeminal nerve REZ and of the brain stem. Consequently, the facial nerve REZ is severely affected leading to the presence of tic convulsif in TN caused by TVBS. PMID- 9217417 TI - [Automatic movements of extremities induced in primary massive brain lesion with apneic coma]. AB - In three cases of primary massive brain lesion with apneic coma, various automatic movements of the extremities were elicited by physical or sensory stimulation. In each case, these movements appeared after a period of cessation of spontaneous respiration followed by flaccid tetraplegia. Brainstem reflexes were absent throughout in all cases. The movements were induced mainly by ventroflexion of the neck, and each case showed movements as described below: in the first case, the patient flexed her elbows and raised both arms slowly, a typical Lazarus sign; in the second case, the patient raised both arms and showed myoclonic movements; and the third case showed abduction of both legs and extension in the upper extremities. Pathology in the first case showed ischemic changes in the entire brain and brainstem. Although ischemic change was also found in the anterior horn cells and white matter of the spinal cord of C1-C4 and of T4 and below, the spinal cord of C5-T3 was relatively well-preserved. These movements appear to have essentially originated in spinal neurons; however, it is assumed that they must have recovered from spinal shock which occurred due to upper level transection. These movements were induced by ventroflexion of the neck, so mechanical extension of the spinal roots, mechanical compression of the spinal cord, and various modalities of the sensation afferent might have some relation to these movements. As tonic neck reflex might also be a cause of these, movements, association with the lower medulla could not be ruled out completely. These movements appeared nearing or after brain death. Although in each case of brain death the spinal cord may have been affected by specific conditions, such as impaired circulation of whole central nervous sysytem, it might have been transversed at upper level, which then causes spinal automatism. These movements might appear even in the state of brainstem death. In each case, the distribution and severity of hypoxic changes in the spinal cord may have resulted in variations in the type and characteristics of these movements. PMID- 9217418 TI - [Clinical characteristics and muscle histopathology in polymyositis positive anti hepatitis with C virus antibody]. AB - In 14 patients with polymyositis (PM), 5 patients (2 males and 3 females) were positive for anti-hepatitis C virus (HCV) antibody measured by a second generation assay. We analysed the clinical characteristics and histopathological findings of the biopsied muscles from those 5 patients. They aged from 42 to 65 years averaging 53.6 years. Two asymptomatic patients visited our hospital due to elevated muscle enzyme levels, who had slight weakness in their orbicularis oculi and neck muscles on physical examination. The other 3 patients had moderate weakness of the proximal muscles. Anti-nuclear antibody was positive in 2 of the 5 patients and anti-Jo 1 antibody was negative in all patients. The serum enzymes elevated were creatine kinase (215-2, 207 (IU/l)) and glutamate oxaloacetate transaminase (40-119 (KU)). HCV-RNA was positive in the sera of 4 patients examined. All muscle biopsy specimens revealed variation in fiber size with inflammatory cellular infiltration and observed degenerating and regenerating fibers. The scant infiltration type was observed in 2 asymptomatic patients in whom the infiltrated cells were CD4 positive. The endomysial infiltration type was observed in 3 symptomatic patients; CD8 positive cells were found focally to diffusely in 2 patients examined. The expression of class 1 molecules from the major histocompatibility complex was detected mainly in infiltrated fibers to variable degrees. All of the patients showed a good response to the initial steroid therapy. The present study suggests that autoimmune reaction related to HCV infection causes myositis, therefore anti-HCV antibody should be checked in cases of PM. PMID- 9217419 TI - [Difference of new mutation rates in dystrophin gene between deletion and duplication mutation in Duchenne and Becker muscular dystrophy]. AB - To clarify new mutational rates in the dystrophin gene between deletion and duplication mutations, carrier diagnosis was performed on 123 mothers of probands suffered from Duchenne (DMD) and Becker (BMD) muscular dystrophy. Quantitative Southern blot analysis with cDNA probes was applied in this study. Out of 108 mothers of DMD/BMD patients with deletion mutation in dystrophin gene, 69 were carriers and 39 were non-carriers. On the other hands, all of 15 mothers of probands with duplication mutation were carriers. The fact that no new mutation occurred in oogenesis in the families with duplication mutations in dystrophin gene indicates that duplications arise in spermatogenesis. The risk of the mother of an isolated case of DMD/BMD with duplication mutation of being a carrier is significantly higher than the estimated risk based on the equality of new mutation in oogenesis and spermatogenesis. PMID- 9217420 TI - [Acute acalculous cholecystitis as a complication of cerebrovascular disease]. AB - Acute acalculous cholecystitis (AAC) is a potentially life-threatening complication, which is sometimes found in patients with multiple injuries, burns, or after an operation. It is unclear, however, whether AAC occurs after cerebrovascular disease (CVD). We studied the incidence of AAC complicating CVD and the clinical characteristics of AAC that occurs after CVD. One thousand three patients with CVD were studied who had been admitted at the acute stage to Kenwakai Hospital from January 1989 through September 1995 and to Seguchi Hospital of Neurosurgery from January 1993 through September 1995. There were 557 patients with cerebral infarction, 273 with cerebral hemorrhage, 94 with subarachnoid hemorrhage, and 79 with TIA/RIND. Twelve patients developed acute cholecystitis, ten of whom had AAC. Of the ten patients with AAC, six had cerebral infarction, two cerebral hemorrhage, and two TIA/RIND. Eight of ten were male. The incidence of AAC was 1.0% in the CVD patients studied. The majority of the AAC patients showed severe hemiparesis. The time interval from CVD to the onset of AAC ranged from 1 to 89 days, with a mean of 25.1 days. AAC occurred 0 to 16 days (mean 5.8 days) after the start of oral or tube food intake in five patients. The most common initial symptom was fever (70%), whereas abdominal pain was infrequent (20%). All the patients showed elevated CRPs and abnormal ultrasonographic findings for the gallbladder and some also had leukocytosis (60%) and elevated aminotransferase of more than 100 IU/l (30%). Cholecystectomy was performed on four AAC patients, but five were successfully treated with antibiotics. The cause of AAC complicating CVD seems to be multifactorial and probably is related to fasting, increased bile concentration, and arteriosclerosis. Our results strongly suggest that AAC is an unrecognized but important complication during acute stage CVD patients. PMID- 9217421 TI - [Axonal motor neuropathy associated with anti-SGPG antibody]. AB - We report a case of axonal motor neuropathy associated with anti-sulfated glucuronic paragloboside (SGPG) antibody which has not been reported yet. A 49 year-old man was admitted with asymmetrical patchy weakness which started in his upper extremities. The deep tendon reflexes were absent in the upper extremities, but normal in the lower extremities. There was a slight decrease in the vibratory sensation in the distal portions of the lower extremities. The general laboratory tests including the protein level of the cerebrospinal fluid revealed no abnormalities. Motor conduction studies showed the low amplitudes of CMAP and no demonstrable conduction block in the limbs. Sensory conduction studies showed no abnormalities except for slightly decreased amplitude of SNAP in the limbs. Electromyography showed active denervation in the upper extremities. Serial electrophysiological studies suggested that the predominant process was axonal degeneration of the motor nerves. Thin-layer chromatography with immunostaining showed that his serum IgM reacted with SGPG. Treatment with cyclophosphamide and corticosteroids was unsuccessful. In this case, this anti-SGPG antibody may be involved in the pathogenesis of chronic axonal degeneration of the motor nerves. PMID- 9217423 TI - [The diagnosis of amyotrophic lateral sclerosis supported by motor evoked potential and brain MRI studies]. AB - A 57-year-old man developed severe muscle weakness and atrophy of the upper extremities within a five-month period. Neurological examination revealed severe weakness and atrophy in the scapular muscles and proximal and distal muscles of the upper extremities. Fasciculations were also observed in the various muscles of the upper extremities. There was neither muscle weakness, atrophy nor fasciculation in either his face, neck muscles or lower extremities. He had no pseudobulbar or bulbar signs. Tendon reflexes were mildly hyperactive in the jaw and lower extremities, and normal in the upper extremities. There were no pathological reflexes, spasticity or sensory disturbances. The needle EMG study revealed denervation potentials in all muscles of the upper extremities examined. The nerve conduction study revealed no findings of the conduction block. Cervical spine X-rays revealed the narrowing of the spinal foramens at the left C3/C4 and bilateral C4/C5, C5/C6, and C6/C7 intervertebral levels. In addition, magnetic resonance imaging (MRI) revealed compressions of the cervical cord at C4/C5 and C5/C6 intervertebral levels. These clinical and neuroradiological findings resembled those of the cervical spondylotic amyotrophy (CSA). However, the motor evoked potential (MEP) study revealed the pyramidal tract dysfunction above the levels of the pyramidal decussation. Furthermore, brain MRI revealed abnormal foci in both internal capsules which were characterized by hyperintense relative to cortical gray matter on T2-weighted images and still hyperintense to white matter on proton-density-weighted images. In addition, T2-weighted images demonstrated a low signal within the motor cortex and hyperintense lesions in the white matter of the precentral gyri. These MRI findings indicated the degeneration of the pyramidal tract and corresponded to those found in the patients with amyotrophic lateral sclerosis (ALS) which have been recently reported. It has been difficult to distinguish ALS from CSA. However, MEP and brain MRI studies were useful for distinguishing these two diseases in this patient. In addition, this patient showed typical MRI findings suggesting the degeneration of the pyramidal tract, although this patient had a relatively short course of illness and did not show obvious physical findings suggesting pyramidal tract dysfunction. PMID- 9217422 TI - [A case of chronic inflammatory demyelinating neuropathy associated with antibodies to gangliosides GM1 and GalNAc-GD1a]. AB - In January 1993, a 43-year-old man was admitted to our hospital for left wrist drop. Neurological examinations revealed asymmetrical distal weakness in the upper limbs. Deep tendon reflexes were normal in all 4 limbs. Sensory and autonomic nervous functions were intact. CSF examinations were within normal limits. Thin-layer chromatography with immunostaining revealed serum antibodies that reacted with GM1 and GalNAc-GD1a. Motor nerve conduction studies revealed abnormal temporal dispersion, and a low amplitude of compound muscle action potential in the left radial nerve. Neurological symptoms gradually improved with prednisolone over one and a half years. He was hospitalized again in January 1995, because of right wrist-drop and slight sensory loss of the limbs. Those findings were improved by methylprednisolone (1,000 mg/day) for 3 days. The interval until maximal disability in this patient was more than one month for each admission. This case must belong to inflammatory demyelinating neuropathy. PMID- 9217424 TI - [A case of potassium-sensitive periodic paralysis with cardiac dysrhythmia]. AB - The authors reported a case of potassium-sensitive periodic paralysis with cardiac dysrhythmia. The patient was a 21-year-old male and had periodic paralysis and asymptomatic cardiac dysrhythmia since the age of 12. His attacks worsened in frequency and intensity which brought him to our hospital at the age of 21. Physical examination on admission revealed slight dysmorphic features such as hypoplastic mandible and high-arched palate. He had slight proximal muscle atrophy with no myotonia. Electrocardiogram showed multifocal ventricular arrhythmia. The serum potassium levels during his paralytic attacks were normal or slightly decreased (3.6-4.2 mEq/l). Both potassium and glucose tolerance tests provoked paralytic attacks. Glucose tolerance test also aggravated his cardiac dysrhythmia. Acetazoramide administration improved his paralytic attack. Potassium-sensitive periodic paralysis with cardiac dysrhythmia can not be defined by the classification of periodic paralysis based on the serum potassium concentration. Provocative tests should be done to make a definite diagnosis and treatment should be done taking into consideration both paralytic attack and cardiac dysrhythmia. PMID- 9217425 TI - [Relapsing polychondritis with mental disorders: a case report]. AB - A 60-year-old man developed disorientation, euphoria, hyperactive behavior, mental confusion, and a moderate decrease of cognition during the course of relapsing polychondritis with bilateral auricular chondritis, sensorineural hearing loss, episcleritis and nasal chondritis. An electroencephalogram showed diffuse slow wave abnormality. The CSF analysis revealed pleocytosis and increased protein. MRI with contrast showed enhancement and edema in bilateral medial temporal regions. These abnormal findings improved markedly with the treatment of corticosteroids. This is the first Japanese case report of relapsing polychondritis presenting as mental disorders. PMID- 9217426 TI - [A case of corticobasal degeneration presenting with primary progressive aphasia]. AB - We report a 64-year-old right-handed woman whose initial symptom was slowly progressive aphasia without generalized dementia and who was subsequently diagnosed as having corticobasal degeneration (CBD). Neurological examination revealed disturbed vertical gaze, dysarthria, rigidity of the right upper extremity, and bilateral instinctive grasp reaction. Neuropsychological assessment disclosed Broca's aphasia, buccofacial apraxia, and memory disturbance. MRI of the brain showed atrophy of the frontotemporal lobes, which was more severe on the left than on the right, especially the left inferior frontal gyrus. In most reported cases of CBD, the initial symptom is motor dysfunction of the unilateral upper or lower extremity. However, we should be cautious that among cases with CBD, there have been rare cases that begin with progressive aphasia alone. In our case, the atrophied region of the cerebral cortex was most severe around the left inferior frontal gyrus. In a few reported cases with the initial symptom of aphasia, the atrophied region corresponded considerably to the type of the aphasia. On the other hand, in those whose initial symptom was mainly motor dysfunction of the unilateral extremity, the atrophied region was remarkable in the posterior part of the frontal lobe and parietal lobe. Therefore, we suggest that in CBD the distribution of the cerebral cortical lesions differs in accordance with whether the initial symptom is motor disturbance or aphasia, and that the type of aphasia corresponds to the location of the cortical lesion. PMID- 9217427 TI - [A case of vertebral artery dissection with lateral inferior pontine syndrome and lateral medullary syndrome]. AB - A 55-years-old woman had left neck pain and headache, dizziness, left Horner's sign, left abducens palsy, diplopia, left peripheral facial palsy, left loss of hearing, left tinnitus, left paralysis of vocal cord and soft palate, dysphagia, left limb ataxia, truncal ataxia, disturbance of temperature and pain sensation over Th10 on the right involving the right face. Left vertebral angiography revealed tapering occlusion of the left vertebral artery. Right vertebral angiography showed normal angiogram of the basilar artery and bilateral anterior inferior cerebellar arteries. MRI disclosed infarcts in the left lateral inferior pons, left lateral medulla, and cerebellum of territories in the anterior inferior cerebellar artery and posterior inferior cerebellar artery. T2 weighted image showed septum (intimal flap) in the left vertebral artery. This is the very rare case of lateral inferior pontine syndrome and lateral medullary syndrome due to the vertebral artery dissection. PMID- 9217428 TI - [A case of left subclavian artery occlusion with transient ischemic attacks probably in the internal carotid artery system]. AB - The subclavian steal syndrome is known to steal blood flow from the vertebrobasilar system. However, we experienced a case of subclavian artery occlusion presenting transient ischemic attacks in left internal carotid system. A left handed 41-year-old man developed transient dysarthria and right hemiparesis including face several times when he physically used his arms. He had no symptoms of the vertebrobasilar system. A brain MRI revealed an old cerebral lacuna at the left putamen supplied by perforating arteries of the middle cerebral artery. The angiography demonstrated a complete occlusion of the proximal portion of the left subclavian artery without a reverse flow from the vertebral artery. Instead, descending cervical branches and deep cervical branches of the ipsilateral external carotid artery supplied collateral pathways to the occluded subclavian artery. On the basis of above observations, we speculated that he developed symptoms of the internal carotid system due to the steal through the collateral network of the cervical arteries directed to the subclavian artery. We should consider not only the vertebrobasilar system but also the internal carotid system, especially its cervical artery network, when exploring collateral pathways for the subclavian steal syndrome. PMID- 9217429 TI - [Sibling cases of opsoclonus-polymyoclonus syndrome]. AB - We described a 34-year-old man who developed opsoclonus-polymyoclonus syndrome (OPS) associated with benign encephalitis. His older sister also suffered from the same syndrome 12 years ago. We examined HLA types in 6 patients with OPS who were admitted to our hospital, including these sibling cases. All 6 patients have type A2 antigen and statistical analysis suggested that there is a positive relationship between OPS and HLA A2 antigen. Considering the two facts that the existence of the sibling cases in this relatively rare disorder and sharing of HLA A2 antigen in our 6 cases, some genetic factors might be involved in the development of OPS. PMID- 9217430 TI - [L-dopa/benserazide during pregnancy in a patient with juvenile parkinsonism]. AB - We report a 34-year-old woman with juvenile parkinsonism who had been treated with five tablets daily of l-dopa/benserazide (100 mg/25 mg). She became pregnant in May 1995 and continued the same treatment. She delivered a healthy boy (3,798 g, 52 cm) with Apgar scores of 9 at 38 weeks gestation by normal delivery. In animal study, there is a report of teratogenicity (bone abnormality) of l dopa/benserazide in rabbits at doses of 30 mg/kg and 120 mg/kg. However, there have been several reports of uneventful delivery of normal children in patients with juvenile parkinsonism who had been treated with l-dopa/decarboxylase inhibitor (DCI) during pregnancy. There seems to be no firm reason to avoid pregnancy during the treatment of l-dopa/DCI or to withhold l-dopa/DCI during pregnancy. PMID- 9217431 TI - [Secondary parkinsonism following midbrain hemorrhage]. AB - We report a patient who developed right sided cogwheel rigidity and resting tremor after left midbrain hemorrhage. Brain magnetic resonance imaging (MRI) showed left midbrain old hemorrhage including substantia nigra. I-123 iodoamphetamine single photon emission computed tomography (IMP-SPECT) images showed reduced radioisotope (RI)-uptake in the left striatum, thalamus and frontal lobe. Our report shows that focal midbrain lesion can produce parkinsonism. PMID- 9217432 TI - Potential health economic benefits of vitamin supplementation. AB - This study used published relative risk estimates for birth defects, premature birth, and coronary heart disease associated with vitamin intake to project potential annual cost reductions in U.S. hospitalization charges. Epidemiological and intervention studies with relative risk estimates were identified via MEDLINE. Preventable fraction estimates were derived from data on the percentage of at-risk Americans with daily vitamin intake levels lower than those associated with disease risk reduction. Hospitalization rates were obtained from the 1992 National Hospital Discharge Survey. Charge data from the 1993 California Hospital Discharge Survey were adjusted to 1995 national charges using the medical component of the Consumer Price Index. Based on published risk reductions, annual hospital charges for birth defects, low-birth-weight premature births, and coronary heart disease could be reduced by about 40, 60, and 38%, respectively. For the conditions studied, nearly $20 billion in hospital charges were potentially avoidable with daily use of folic acid and zinc-containing multivitamins by all women of childbearing age and daily vitamin E supplementation by those over 50. PMID- 9217433 TI - Geriatrics: the effect of time in medicine. PMID- 9217434 TI - Antiretroviral therapy for human immunodeficiency virus infection in 1997. AB - It has become clear that the acquired immunodeficiency syndrome follows continuous replication of the human immunodeficiency virus (HIV) and a decrease in immune capability, most obviously a decline in the number of CD4 lymphocytes. An understanding of key elements in the infectious life cycle of HIV has led to the development of potent antiretroviral drugs selectively targeting unique reverse transcriptase and protease enzymes of the virus. Completed clinical trials have shown that antiretroviral therapy for HIV infection, begun early, reduces viral replication and reverses the decline in CD4 lymphocyte numbers. Recent studies of combination therapies have shown that decreases in plasma HIV viremia to low levels and sustained increases in CD4 cell numbers are associated with longer survival. Potent combination regimens including protease inhibitors and non-nucleoside reverse transcriptase inhibitors suppress detectable viral replication and have demonstrated clinical benefits in patients with advanced disease. Progress in antiretroviral therapy and methods to monitor responses to treatment are providing new hope in the treatment of HIV infection. PMID- 9217436 TI - Bacterial resistance and the dilemma of antibiotic usage. PMID- 9217435 TI - Who gets a heart? Rationing and rationalizing in heart transplantation. AB - National policy on organ transplantation is made by the United Network for Organ Sharing (UNOS), a representative body composed of health care professionals and patients. Standardized criteria for determining when a patient should be placed on the waiting list for heart transplantation are now in effect nationwide. Current and future directions to maximize the utilization of available donated organs are explored. PMID- 9217437 TI - Advances in the treatment of respiratory failure in newborns. PMID- 9217438 TI - Helicobacter pylori infection: pediatric aspects. PMID- 9217439 TI - Update on childhood asthma. PMID- 9217440 TI - Immunization update. PMID- 9217441 TI - The use of umbilical cord blood stem cells for hematopoietic reconstitution. PMID- 9217442 TI - Attention disorders: the advance is a return to basics. PMID- 9217443 TI - Coccidioidomycosis and sarcoidosis. Multiple recurrences. PMID- 9217444 TI - Diminutive Russian prosthetic heart valve as an iatrogenic cause of mitral stenosis. PMID- 9217445 TI - Gastrointestinal bleeding and gastric outlet obstruction from Peutz-Jeghers polyposis. Diagnosis and treatment. PMID- 9217446 TI - Children, adolescents, and television. A call for physician action. PMID- 9217447 TI - Physician-hastened death. Advisory guidelines for the San Francisco Bay area from the Bay Area Network of Ethics Committees. AB - Recent high court opinions and pending Supreme Court rulings on the legality of physician-hastened death necessitate a pragmatic response from the medical profession. Adopting a "harm reduction" perspective on this contentious topic, the Bay Area Network of Ethics Committees developed practice guidelines for responding to a patient request for hastened death. The guidelines will be offered to the local medical community for use by individuals and health care institutions if the practice of physician-hastened death becomes legal. A multidisciplinary consensus process was used in developing the guidelines, which address clinical, ethical, and procedural concerns. PMID- 9217448 TI - The Northern California Conference for Guidelines on Aid-in-Dying. AB - End-of-life care in the United States is inadequate. Long-standing and unresolved issues in the care of the terminally ill have led to debates that have become major bioethical issues. Recognizing that practical solutions to deal with these issues are desperately needed, the Stanford University Center for Biomedical Ethics organized and convened a consensus development conference for health care professionals and health care institutions on Sept. 27 and 28, 1996. PMID- 9217449 TI - Report of the Northern California Conference for Guidelines on Aid-in-Dying: definitions, differences, convergences, conclusions. AB - In September 1996, the Stanford University Center for Biomedical Ethics convened a conference entitled "Comprehensive Care of the Terminally Ill: The Northern California Consensus Development Conference for Guidelines on Aid-in-Dying." The regionally based, multidisciplinary conference gathered people from a variety of disciplines and diverse perspectives on physician aid-in-dying. This report documents important points of convergence, disagreement, and uncertainty that emerged from the conference and provides commentary on crucial issues: the definition of terminal illness, ensuring adequate palliative care, psychiatric challenges, coping with family pressures, the doctor-patient relationship, the managed care context, the role of ethics committees, and institutional challenges. Should physician aid-in-dying become a legal practice in California, the report will provide guidance to health care organizations, health professionals, and public policy officials engaged in local or state guideline or policy development. PMID- 9217451 TI - Physician-assisted suicide. Finding common ground. AB - In Washington state, practicing physicians have been forced to confront the emotional, complex issue of physician-assisted suicide sooner than physicians elsewhere in the US. The Washington State Medical Association has struggled at length with the issue and ultimately delineated a policy on safeguards for physician-assisted suicide. The Washington experience may prove instructive to other professional physician organizations even before the US Supreme Court rules on the issue. PMID- 9217450 TI - Futile care policy. Lessons learned from three years' experience in a community hospital. AB - After reviewing intensive and life supportive care for patients in whom it was probably inappropriate, the hospital bioethics committee established a futile care policy. The issues we encountered in the first 3 years since the policy became effective should be instructive for other hospitals, physicians, and families. PMID- 9217452 TI - The Oregon Death With Dignity Act: implementation issues. AB - Passage of the Oregon Death With Dignity Act in 1994 raises nationally relevant questions for health care organization, state agencies, and clinicians. As debate over physician-assisted suicide continues in the United States, the experiences in Oregon may offer insight into the clinical complexities of legalizing physician-assisted suicide. PMID- 9217453 TI - Physician-assisted suicide. Overview of the ethical debate. PMID- 9217454 TI - Protecting the innocents. People with disabilities and physician-assisted dying. PMID- 9217455 TI - Is the slippery slope steeper for people with disabilities? PMID- 9217456 TI - Increasing the brain's tolerance to ischemia in the operating room. PMID- 9217457 TI - Recent advances in the management of spinal cord injury. PMID- 9217458 TI - Management of internal carotid artery occlusion. PMID- 9217459 TI - Traumatic brain injury. PMID- 9217460 TI - Clostridium perfringens outbreak at a juvenile detention facility linked to a Thanksgiving holiday meal. PMID- 9217461 TI - Reactive arthritis to Clostridium difficile in a child. PMID- 9217462 TI - Heart rate response to industrial work at different outdoor temperatures with or without temperature control system at the plant. AB - Different outdoor temperatures, the association between indoor temperature control at the workplace and working heart rates of industrial employees were evaluated. The subjects, 6,016 male and female employees in 21 industrial plants in Israel, were screened for cardiovascular risk factors between 1985-87 (The CORDIS Study). The data collected included resting heart rate, working heart rate (based on one hour ambulatory ECG), outdoor temperatures, temperature control (TC) status of the plant, workload, age and health-related habits. At outdoor temperatures below or above 22-28 degrees C, subjects working in plants with TC had lower mean working heart rate HR (-2 bpm) than those working in plants without TC (p < 0.0004 after adjustment for confounders). No statistically significant differences in mean working HR were found between subjects working with TC (at all the outdoor temperatures) and those without TC within the outdoor temperature range 22-28 degrees C. Based on working heart rate, indoor temperature control in industrial plants appears to moderate the cardiovascular strain in working subjects during both cold and hot days. PMID- 9217463 TI - The baculovirus 10-kDa protein. PMID- 9217465 TI - Justify this. PMID- 9217464 TI - Intramolecular proteolytic cleavage of Bacillus thuringiensis Cry3A delta endotoxin may facilitate its coleopteran toxicity. AB - The Cry3A delta-endotoxin protein inclusion synthesized by Bacillus thuringiensis subsp. tenebrionis is soluble in alkaline and acid buffer solutions but the toxin precipitates when returned to neutral pH conditions. The midgut pH of susceptible beetle larvae is neutral to slightly acidic, a pH environment in which the Cry3A toxin is insoluble. To investigate this paradox we studied the Cry3A toxin after various proteolytic treatments. In many cases the toxin was cleaved into polypeptides that remained associated under non-denaturing conditions. Interestingly a chymotrypsinized Cry3A product was soluble under neutral pH conditions, retained full activity against susceptible beetle larvae, and exhibited specific binding to Leptinotarsa decemlineata midgut membranes. PMID- 9217466 TI - Inefficiency experts? PMID- 9217467 TI - Managed care. Up and away. PMID- 9217468 TI - Education. Crowded fields. PMID- 9217469 TI - Quality watch. Physician, accredit thyself. PMID- 9217470 TI - Quality watch ... looking for ways to measure your health plan or network? PMID- 9217471 TI - Litigation. Shady operations. PMID- 9217472 TI - Compensation ... hospital executives. PMID- 9217473 TI - Home health ... if your home health agency has been good this year. PMID- 9217474 TI - Mergers/acquisitions ... health care not-for-profits in Columbia, S.C. PMID- 9217475 TI - Leadership ... chief executives nearing retirement. PMID- 9217476 TI - Public health ... people are looking to their own neighborhoods. PMID- 9217477 TI - Human resources ... hospitals are less likely to fire employees this year. PMID- 9217478 TI - Policy ... limiting acute care for the very old at the end of their lives saves tons of money. PMID- 9217479 TI - Liability ... review of radiology-related malpractice claims. PMID- 9217480 TI - Second thoughts on selling. Interview by Chris Serb. PMID- 9217481 TI - Capital: who's got it? How to get it! AB - Cruel. Fickle. Demanding. While hospitals and HMOs face tougher scrutiny by credit-rating agencies, Wall Street has slowed the flow of capital to practice management and assisted-living start-ups. Meanwhile, some investor-owned hospitals now emphasize debt over equity. In this special report, we examine the capital markets sector by sector. PMID- 9217482 TI - Foundations. Show us the money. AB - Sales of hospitals to for-profit chains have spawned nearly 100 new community foundations. Consumers and state officials increasingly want to know: How will these new charities spend their funds. PMID- 9217483 TI - The new VA. AB - Once the epitome of bloat, the Veterans Administration has slashed and slimmed its health care operations. The tactics mirror the marketplace: regional networks, consolidated purchasing, more outpatient care, capitation-like payment rates, and stiff performance targets. VA execs say it may actually work. PMID- 9217484 TI - The good death. The people of Missoula, Montana, open a dialogue on better ways to die. AB - Missoula, Mont., is abuzz with talk about dying. A long-term project, the brain child of a local hospice doctor, is fostering a community dialogue on better ways to die. Hospitals, nursing homes and others have joined in. PMID- 9217485 TI - Consumers. The gate swings wider. PMID- 9217486 TI - Disability benefits. Testing the limits. PMID- 9217487 TI - Alliances. Survival of the biggest. PMID- 9217488 TI - Policy. Up in smoke. PMID- 9217489 TI - Information systems. A hospice goes high-tech. PMID- 9217490 TI - Outreach. Health care spoken here. PMID- 9217491 TI - Assisted living. Shaking hands for a change. PMID- 9217492 TI - Cardiac care. A line drawn in the sand. PMID- 9217493 TI - More on "right-sizing residencies". PMID- 9217494 TI - More on "right-sizing residencies". PMID- 9217495 TI - More on "right-sizing residencies". PMID- 9217496 TI - More on "right-sizing residencies". PMID- 9217497 TI - Reassessing the OSCE. PMID- 9217498 TI - Reassessing the OSCE. PMID- 9217499 TI - IMGs and cultural diversity training. PMID- 9217500 TI - Chart review for residents. PMID- 9217501 TI - A progressive geriatrics curriculum. PMID- 9217502 TI - Medical universities and academic health centers: lessons of history. PMID- 9217503 TI - Med-peds--three decades of the generic primary care physician. PMID- 9217504 TI - The future supply of physicians. AB - Many health services researchers point to a growing surplus of physicians by the end of the century. The author discusses in detail a variety of policy positions, from the Flexner Report onward, that have affected the present and projected supplies of U.S. physicians. These include the American Medical Association's decades of efforts to control the numbers and types of U.S. medical students; effects of Medicare and Medicaid; changes in immigration and naturalization laws that increased the number of international medical graduates (IMGs); the medical community's non-response to the 1981 GMENAC Report's forecasts on physician oversupply; growth in the numbers of specialists; the fall and subsequent rise in the numbers of applicants to medical schools; the changing composition of the physician workforce; the refusal of the medical profession to consider a shorter training period for physicians; and other events from the past that can inform today's policymakers. The author then evaluates four policy recommendations that have evolved to deal with the problem of physician oversupply, and concludes that (1) reliance on the market to contain physician supply is unwarranted; (2) there is little prospect that Congress will soon reduce the inflow of IMGs, and even if it did, such action would have a marginal effect; (3) there is no prospect that 20-25% of U.S. medical schools will be closed by 2005, since the forces militating against such action are overwhelming; and (4) it remains to be seen whether the new health care environment will have more than a marginal effect in altering the current ratio of primary care to specialist physicians in the years ahead. In fact, if future outlays for health care increase as predicted, there should be sufficient funds for physician supply to continue to grow and for specialists to continue to make good incomes. PMID- 9217505 TI - The continuing dilemma in clinical investigation and the future of American health care: a system-wide problem requiring collaborative solutions. AB - American medicine faces a paradox: on the one hand, decades of basic science research have produced fundamental insights into disease mechanisms. On the other, there has rarely been a more difficult environment for the training and employment of clinical investigators, who perform the research that translates basic biomedical knowledge into practical advances in patient care. The authors explain the historical roots of this crisis and the lack of data about specific workforce needs for clinical investigators, discuss long-standing difficulties of recruiting, training, and retaining these scientists (e.g., time-consuming training, inadequate emphasis on clinical research in medical school, fewer role models) and why these processes are becoming more difficult (e.g., a coming flattening of federal support for research; the impact of managed care on academic health centers). In confronting this problem, the authors stress the importance of (1) carefully defining the major subtypes of clinical investigation (i.e., physiologic investigation, outcomes research, and clinical trials) and noting which are and which are not endangered; (2) understanding the potential solutions to the problem that have been offered in the past; and (3) defining and hopefully marshaling a coalition of those institutions whose resources are currently available to address the problem and that have an important stake in its solution: academic health centers, the National Institutes of Health, health care providers, foundations and educational societies, medical schools, and industry. The authors stress that finding solutions to the current problem are a shared responsibility that must be carried out, for without well-trained and innovative clinical investigators, the social contract of biomedical research-to keep society well-cannot be fulfilled. PMID- 9217506 TI - Importance and difficulty of determining the cost of clinical research. AB - The cost of clinical research has become increasingly important over the last few years because of changes in the health care market and the resulting decline in hospital margins. The emergence of contract research organizations has also caused academic medical centers to pay attention to costs. Although the consideration of clinical research costs has become vital, a number of factors make cost evaluation a difficult process. These include confusion about terminology, lack of specificity, estimation of the cost of complications, and duration of patient participation, as well as the assignment of personnel costs and infrastructure costs. PMID- 9217508 TI - The teaching matrix: a tool for organizing teaching and promoting professional growth. AB - Despite barriers of limited time and lack of formal preparation for teaching, physician-teachers want to do a good job in the classroom. However, without appropriate feedback or self-reflection, physician-teachers have no systematic way to think about their role both in what students learn and in how well they understand important information. With this in mind, the authors developed a model, the Teaching Matrix, designed to encourage clinician-teachers to reflect on their teaching before, during, and after each teaching session. The Matrix helps teachers to address five central questions: Who am I teaching? What am I teaching? How will I teach it? How will I know if the students "got it"? And how will I improve my teaching for the next time? In this paper, the authors describe how the Teaching Matrix may be used as a tool for planning actions, as a "suggestion box" of ideas, advice, and questions, and, most importantly, as a guide to systematic reflection on teaching. PMID- 9217507 TI - Preserving medical schools' academic mission in a competitive marketplace. AB - To gain a better understanding of the effects on medical schools of transformations in medical practice, science, and public expectations, the AAMC in 1994 formed the Advisory Panel on the Mission and Organization of Medical Schools and appointed six working groups to address relevant issues. This article is a report of the findings of the Working Group on Preserving Medical Schools' Academic Mission in a Competitive Marketplace, which was charged with exploring how medical schools could acquire and/or preserve an adequate patient base for teaching, research, and income generation in a competitive marketplace. The other groups' reports will appear in future issues of Academic Medicine. To understand the diversity of approaches that schools have taken to achieve this goal and to preserve their missions, the group interviewed representatives of nine medical schools, selected to represent a cross section of U.S. medical schools. The interviews took place on four occasions between June 1995 and March 1996. The information and comments shared by participants helped the working group gain insight into the fundamental issues it had been charged to address, including those of new delivery structures, what value schools offer to delivery structures, how education and research can be incorporated and supported financially, possible new pressures on relationships between medical schools and teaching hospitals, changes in faculty physicians' employment relationships and terms, and the role of the medical school in graduate medical education. In collecting and analyzing the data, the working group focused on the distinction between protecting an institution's existing enterprise and preserving an institution's core mission. This article gives a detailed overview of the information and comments each school presented, organized under the appropriate question. The working group's conclusions and commentaries on the findings follow. An appendix presents more detailed summaries of the schools' presentations, organised as case studies. The picture that emerges is complex. The working group concluded that medical schools will take a variety of approaches to define and preserve their missions. Most, but not all, medical schools will be able to secure the patient bases necessary to fulfill their missions even in a competitive marketplace. However, the nature of many of the schools is likely to change, and it is not clear whether the core missions of education and research will continue at their present levels at all schools. PMID- 9217509 TI - Prevention and treatment of alcohol-related problems: an international medical education model. AB - Alcohol abuse and alcoholism are among the world's most pressing public health concerns. Research has shown that while primary care physicians are in a good position to screen for alcohol-use disorders and to aid in treating these problems, they tend to identify only a small percentage of patients with such disorders and they rarely intervene with these persons. This situation is probably attributable to the fact that medical students worldwide are taught very little about alcohol-related problems. Clearly there is an urgent need to educate the world's doctors about preventing, diagnosing, and treating alcohol abuse and addiction. In this paper, the authors describe a model international program for educating physicians about alcohol-related problems that was developed by the National Institute on Alcohol Abuse and Alcoholism (NIAAA) in cooperation with the Center for Addiction Research and Education (CARE) at the University of Wisconsin-Madison. They describe the components of the initiative's "trainer development" approach and critical issues in implementing the program in other countries. Finally, they discuss how the program was successfully implemented in Poland and describe the NIAAA's plans for introducing the model in several other countries. PMID- 9217510 TI - Physics for physicians: integrating science into the medical curriculum, 1910 1950. AB - One of the most difficult problems in twentieth-century medical education has been finding ways to successfully integrate the basic and applied sciences into the medical curriculum. Not only have medical students regarded basic sciences such as physics and biochemistry with distaste, but these subjects traditionally have been taught by pure scientists with little interest in the needs of medical students. In this paper, the author reviews the history of physics teaching at the Queen's University Faculty of Medicine in Canada, placing particular emphasis on the work of J.K. Robertson (1885-1958), professor of physics. Although physics no longer has the relevance to general medical training that it once had a study of Robertson's ideas and methods provides insight into the process of integrating basic science into medical training. Robertson's success in the endeavor was based largely on two factors his "sympathetic understanding" of the needs of medical students and his innovative combination of basic and applied science in one course--factors that are as important to medical teaching today as they were 50 years ago. PMID- 9217511 TI - Medical students as patients: a pilot study of their health care needs, practices, and concerns. AB - BACKGROUND: The personal health experiences of medical students may contribute in important but previously unacknowledged ways to their well-being and education. This pilot study surveyed medical students about their health care needs, practices, insurance status, and concerns about seeking care. METHOD: A questionnaire was developed and distributed to 151 students at the University of New Mexico School of Medicine in 1993-94. Participant privacy was protected. Responses were compiled and analyzed using logistic regression models and odds ratios. RESULTS: A total of 112 students responded. Most reported health care needs and half routinely received care at their training institution. One-third had informally requested prescriptions or diagnostic tests from medical school faculty and housestaff; one-fourth used such informal consultation as their "usual" method of obtaining care. Eighteen students were uninsured. The students reported that they had not sought care for several reasons, and many had experienced difficulty in obtaining care. The students indicated concern about confidentiality and about the dual role as both student and patient at the training institution. They believed that their academic standing would be jeopardized if they developed certain health problems. When asked about hypothetical scenarios, a majority preferred to avoid the dual role of medical student-patient. When asked about scenarios in which medical student peers exhibited suicidal depression or severe drug abuse, the students overwhelmingly preferred not to notify the medical school administration. Significant differences in responses were found with respect to gender and training level. CONCLUSION: This pilot study examined the health care needs, practices (including the use of informal consultation), insurance status, and concerns of students at one medical school. The findings highlight the students' perceptions of illness and vulnerability during medical school training. Constructive implications for academic medicine are discussed regarding initiatives in the areas of policy, research, and the resources and structure of student health care services. PMID- 9217512 TI - Faculty attitudes and opinions about problem-based learning. AB - BACKGROUND: Systematic research on faculty attitudes toward problem-based learning (PBL) has focused exclusively on the opinions of tutors. The purpose of the present study was to examine the attitudes of faculty at a single medical school who either (1) did not participate in the first year of a new PBL curriculum or (2) participated in ways other than as PBL tutors. METHOD: In 1993 94, at the end of the first year of a new PBL curriculum, a questionnaire used in an earlier, larger study of PBL tutors was sent to all 494 faculty at the University of Missouri-Columbia School of Medicine. RESULTS: The response rate was better for participants (76%, 115 of 151) than for non-participants (28%, 96 of 343). Overall, nonparticipants judged the new curriculum to be approximately equal to the "old" curriculum that preceded it. In contrast, participants were significantly more positive and judged the new PBL curriculum to be superior in most respects. For both groups the new curriculum received its highest ratings in the areas of student interest, clinical preparation, and medical reasoning and its lowest ratings in the teaching of factual knowledge in the basic sciences and efficiency of learning. The attitudes of participating faculty varied with their teaching roles in the new curriculum. Those whose primary roles were as PBL tutors or as leaders of other small discussion groups were more favorable to the new curriculum than those who primarily served as lecturers. Faculty who served in several different roles were more positive than faculty who served in only one role. There were also plausible qualitative differences among the teaching-role groups in what they liked and disliked about the new curriculum. CONCLUSION: In general, the attitudes and opinions of the faculty varied with the degrees and types of participation in the new curriculum. The attitudes and opinions of faculty with different teaching roles were plausibly related to differences in these roles. The opinions of the faculty about the strengths and weaknesses of PBL that emerged in this survey are much like those found in prior research. In addition, student performances on Step 1 of the United States Medical Licensing Examination suggest that the faculty may have underestimated the value of the new PBL curriculum for helping students acquire factual knowledge in the basic sciences. PMID- 9217513 TI - Pelvic examination instruction and experience: a comparison of laywoman-trained and physician-trained students. AB - PURPOSE: To evaluate medical student performances of pelvic examinations after completion of the obstetrics-gynecology (ob-gyn) clinical clerkship in order to compare the effectiveness of training by laywomen serving as both teachers and patients with the effectiveness of training by an attending physician as teacher, with a lay-woman serving only as the patient. The study also examined whether students were given additional training and opportunities for practice during their clinical clerkships in other disciplines. METHOD: Following completion of their ob-gyn clerk-ships in 1993 and 1994, a total of 81 students at two North Carolina medical schools answered a questionnaire eliciting demographic information, pelvic examination experience, and the content of the training they had received. The students then performed a pelvic examination on a standardized patient (SP). Their performances were evaluated by the SP using a 35-item scale, subdivided into technical and interpersonal skills. The data were analyzed by two tailed t-tests, analysis of variance, and chi-square tests. RESULTS: The laywoman trained students demonstrated better interpersonal skills than did the physician trained students (p = .01). No significant difference was found in technical skills. Nearly one-fourth of the students reported that communication skills had not been taught during their ob-gyn clerkships. The students reported performing pelvic examinations often on their ob-gyn rotation but infrequently on other rotations. CONCLUSION: The authors recommend that teaching by laywomen be incorporated into the teaching of pelvic examinations and other aspects of a women's health curriculum. Interpersonal skills taught by laywomen in preclinical courses on pelvic examination may have a lasting effect that can be demonstrated after exposure to clinical clerkships. Clinical clerkships should then reinforce these skills. PMID- 9217514 TI - Competing on price: the economics of managed competition. AB - PURPOSE: To describe the economics of teaching hospitals in an increasingly price conscious managed care marketplace by determining the relationships between a teaching hospital's operations and cost per discharge. METHOD: A quantitative correlational regression analysis was undertaken of 1993 operational and financial data from the Health Care Financing Administration for a national sample of 100 major urban, non-federal teaching hospitals. The sample was systematically selected from membership in the Association of American Medical Colleges' Council of Teaching Hospitals. RESULTS: The analysis indicated that the new economics of managed competition requires teaching hospitals to focus on reducing costs through five main areas: decreasing poorly utilized beds, increasing the numbers of discharges, renovating facilities to modernize and streamline patient flow, utilizing fewer employees and thus boosting productivity, and improving the internal financing of operations and investments through working-capital management. CONCLUSION: Achieving efficiency in operations in each of the five main areas will help teaching hospitals to survive the turbulence of market evolution toward managed care. PMID- 9217515 TI - Perceptions of teaching behaviors by primary care and non-primary care residents. AB - PURPOSE: To ascertain the most helpful and least helpful faculty teaching behaviors as perceived by primary care and non-primary care residents and to assess differences in those perceptions between the two resident groups. METHOD: A cross-sectional mailed survey was undertaken in 1993-94 of all 1,046 residents (including interns) in U.S. Army graduate medical education programs. The survey asked the residents to rate the 38 teaching behaviors of Wolverton and Bosworth. Mean ratings were calculated for each teaching behavior, and the ratings of the two resident groups were compared using Kendall's coefficient of concordance. RESULTS: In all, 490 (47%) of the residents responded: 191 (45%) of 421 in primary care, and 299 (48%) of 625 in non-primary care. The primary care and non primary care groups had a high degree of concordance in ranking of the 38 teaching behaviors (W = .953). The exact rank orders differed slightly, but there was disagreement on only one behavior each within the rankings of both the ten most helpful and the ten least helpful behaviors. CONCLUSION: Army residents in all of the major specialties have similar perceptions of what they consider helpful behaviors from their faculty. PMID- 9217516 TI - Reassessing medical students' willingness to treat HIV-infected patients. AB - BACKGROUND: Past research has demonstrated that some physicians do not feel obligated to care for patients infected with the human immunodeficiency virus (HIV). This study sought to characterize the attitudes that affect medical students' willingness to treat HIV-infected patients and to determine which attitudes are most amenable to intervention. METHOD: All 414 matriculating medical students at three Chicago schools were surveyed in 1994. After reliability-testing the attitudinal scales, the authors created a predictive model by using multiple regression analysis. RESULTS: A total of 297 (72%) of the matriculating students responded. Ninety-two percent of the students agreed that patients with HIV would be welcome in their medical practices. As in past studies, a strong sense of professional obligation was associated with willingness to treat, and fear of infection and homophobia were associated with decreased willingness to treat. The authors' measure of concern about social stigma was also associated with decreased willingness. Together, these factors accounted for 53% of the variance in the Willingness to Treat scale. CONCLUSION: In addition to confirming the predictors found in previous studies, this study demonstrated that perceived social stigma is a measurable predictor of decreased willingness to treat (with the understanding that willingness to treat is influenced by both personal and social factors). A comprehensive approach, not only in curriculum design but also in admission and policy, might better prepare students to treat HIV-infected patients. PMID- 9217517 TI - A progress report on accelerated residency programs in family practice. AB - BACKGROUND: In 1991 the American Board of Family Practice (ABFP) approved 12 programs to participate in an experiment in medical education. Selected students in 12 medical schools are able to complete their first year of family practice residency while completing their fourth year of school. This paper reports on the progress of the programs and residents participating in this project. METHOD: Data from the ABFP in-training examination and certification examination were compiled for all trainees and graduates through 1994. Performances were compared with national norms and the performances of traditional residents in the same programs. The program directors were surveyed to assess their experiences, program effectiveness, benefits, liabilities, and implementation problems. RESULTS: Accelerated residents performed better than their peers and national norms on the ABFP in-training and certification examinations. The directors rated the clinical performance of accelerated residents as equal to or better than the clinical performance of traditional residents by the end of the program. Advantages of accelerated residency included improvements in recruiting, image, and morale. Problems occurred in order and prescription writing and acceptance of the accelerated residents by nurses, other residents, and physicians in other disciplines. CONCLUSION: Early entry into residency training of bright, highly motivated, and mature students appears to offer benefits for trainees and programs alike. PMID- 9217518 TI - Organizational models of medical school relationships to the clinical enterprise. AB - The authors analyzed existing relationships between medical schools and clinical enterprises in order to develop models of these relationships. The conceptual framework for the models uses three variables to assess the nature of the relationships: (1) high academic control-high clinical enterprise control; (2) high academic influence-low academic influence; and (3) self-contained system open system (i.e., the extent to which the resources needed for clinical education are provided by the relationship between the clinical enterprise and the medical school). The authors present four conceptual models of the relationship between the medical school and the clinical enterprise: (1) The "single ownership; owned integrated system" is characterized by a closed clinical delivery system owned or controlled by the academic institution. (2) The "general partner" organization emphasizes an open clinical environment in which the medical school forms alliances with clinical entities, and the school is a dominant partner. (3) The "limited partner" organization operates with an open clinical delivery system that the school relates to through affiliations and contractual relationships, and the school is a less dominant partner. (4) The "wholly owned, subsidiary" organization operates in a controlled clinical environment in which the medical school is a subsidiary of the larger integrated delivery system. Each model is presented in its pure organizational form, then augmented with descriptions of the different ways that the medical school and other components may relate to each other. Also, the advantages and disadvantages of each model for the medical school are discussed. The authors emphasize that no model is superior to the others; instead, the best choice for a medical school depends on the history, local circumstances, and leadership of the school and other organizations. The authors' intent is to assist the leaders of medical schools as they design strategies for the future relationships of their institutions. PMID- 9217520 TI - Interhospital patient transfer: a quality improvement indicator for prehospital triage in mass casualties. AB - The need for interhospital patient transfer after mass casualties may be a consequence of triage errors. Indications for interhospital patient transfer following seven suicidal bus bombings in Israel were reviewed to identify possible errors in triage at the scene. Medical records of victims arriving to hospitals were analyzed for age, injury description, injury Severity Score (ISS), and indication and destination of interhospital transfer. A total of 473 victims were involved, 74 of whom died at the scene (15.6%). Mean victim age was 29 +/- 16 (SD) years. Interhospital transfer was necessary for 29 patients. Indications for transfer included (1) mandatory lifesaving procedures on route to trauma center (n = 14), (2) underdiagnosis at the scene (n = 1), (3) insufficient local resources (n = 9), and (4) triage-related errors (n = 5). The ratio between interhospital transfer due to triage errors and the victim population who may need to be transferred is suggested as quality assurance (QA/QI) indicator for triage. PMID- 9217519 TI - Haloperidol, lorazepam, or both for psychotic agitation? A multicenter, prospective, double-blind, emergency department study. AB - Rapid tranquilization is a routinely practiced method of calming agitated psychotic patients by use of neuroleptics, benzodiazepines, or both in combination. Although several studies have examined the efficacy of the three approaches, none have compared these treatments in a prospective, randomized, double-blind, multicenter trial. Ninety-eight psychotic, agitated, and aggressive patients (73 men and 25 women) were prospectively enrolled during an 18-month period in emergency departments in five university or general hospitals. Patients were randomly assigned to receive intramuscular injections of lorazepam (2 mg), haloperidol (5 mg), or both in combination. Patients in each treatment group received 1 to 6 injections of the same study drug within 12 hours, based on clinical need. They were evaluated hourly after the first injection until at least 12 hours after the last. Efficacy was assessed on the Agitated Behavior Scale (ABS), a modified Brief Psychiatric Rating Scale (MBPRS), Clinical Global impressions (CGI) scale, and an Alertness Scale. Effective symptom reduction was achieved in each treatment group with significant (P < .01) mean decreases from baseline at every hourly ABS evaluation. Significant (P < .05) mean differences on the ABS (hour 1) and MBPRS (hours 2 and 3) suggest that tranquilization was most rapid in patients receiving the combination treatment. Study event incidence (side effects) did not differ significantly between treatment groups, although patients receiving haloperidol alone tended to have more extrapyramidal system symptoms. The superior results produced by the combination treatment support the use of lorazepam plus haloperidol as the treatment of choice for acute psychotic agitation. PMID- 9217521 TI - Suicidal ideation in emergency department chest pain patients: panic disorder a risk factor. AB - Most patients who present to the emergency department (ED) for chest pain do not have a cardiac disorder. Approximately 30% of noncardiac chest pain patients suffer from panic disorder (PD), a disabling, treatable, yet rarely detected psychiatric condition. Although still controversial, PD may be a risk factor for suicidal ideation and attempts. The prevalence of recent suicidal ideation (ie, past week) was studied in 441 consecutive ED chest pain patients who underwent a structured psychiatric interview. To examine the controversial link between panic and suicidal behavior, logistic regression analyses were conducted in which current psychiatric diagnoses (Axis I) as well as pertinent medical and demographic information were assessed as risk factors for suicidal ideation. Participants were interviewed with the Anxiety Disorders Interview Schedule Revised to establish psychiatric diagnoses. Recent suicidal ideation (ie, past week) was assessed with question 9 of the Beck Depression Inventory. Ten percent of patients had recent suicidal ideation. Sixty percent of patients with suicidal thoughts met criteria for PD. In the patients with PD, suicidal ideation could not be explained by the presence of comorbid psychiatric or medical conditions or medication. In the total sample, only diagnoses of PD (odds ratio [OR] = 4.3; 95%, confidence interval [CI], 2.09-8.82; P = .0001) and dysthymia (OR = 9.98; 95% CI, 4.00-24.8; P = .00001) were significant and independent risk factors for suicidal ideation. PD, the most common psychiatric condition in ED chest pain patients, may be an independent risk factor for suicidal ideation, further supporting the need for recognition and treatment of these patients. PMID- 9217522 TI - Factors and methodology in achieving ideal delivery temperatures for intravenous and lavage fluid in hypothermia. AB - A study was undertaken to determine the relationship between temperature and delivery rate of warmed intravenous fluid using standard intravenous infusion equipment and tubing. One-liter bags of 0.9% NaCl were warmed to 60 degrees C and run through standard microdrip tubing for 1 hour at rates of 1,000, 800, 600, and 400 mL/h. Thermistor probes were placed into the bag and into the tubing at 0, 100, 180, 230, and 280 cm from the intravenous bag. Separate fluid bags were also warmed to 39.3 degrees and 75 degrees C, and the fluid was run through the same apparatus at 1,000 mL/h and 200 mL/h, respectively. Temperatures were recorded at each site at the start of the infusion and every 10 minutes thereafter for 1 hour, Subsequently, 60-mL syringes of fluid warmed to 39.5 degrees C were eluted through 50 cm tubing over 10 minutes at 300 mL/h and 360 mL/h. Mean delivery temperature over each 10-minute infusion was determined. Fluid preheated to 39.3 degrees C approached room temperature at delivery even at a flow rate of 1,000 mL/h and tubing lengths as short as 100 cm. Fluid preheated to 60 degrees C was delivered at near 37 degrees C using tubing lengths as long as 280 cm when eluted at 1,000 mL/h. Fluid preheated to 39 degrees C in 60-mL syringes and eluted through 50 cm of tubing over a period of 10 minutes at 300 mL/h or 360 mL/h was delivered near a mean temperature of 37 degrees C. These results show that warmed fluid can be delivered through standard intravenous tubing at or near 37 degrees C if the fluid is preheated to 60 degrees C and eluted through long tubing (280 cm) at high flow rates (1,000 mL/h). Alternatively, fluid warmed to 37 degrees C to 42 degrees C can be delivered at or near 37 degrees C via intermittent bolus through short tubing (50 cm) either by hand or syringe pump. The latter approach would be particularly beneficial in the pediatric population, in whom it is not advisable to administer fluid at flow rates as high as 1,000 mL/h. PMID- 9217523 TI - False-positive preliminary radiograph interpretations in a pediatric emergency department: clinical and economic impact. AB - A prospective, case control study at a university-affiliated, academic pediatric emergency department was undertaken to determine the clinical impact and cost of false-positive preliminary radiograph interpretations and to compare the cost of false-positive interpretations with the estimated cost of a 24-hour on-site pediatric radiologist. Data were collected on all patients undergoing radiography of the chest, abdomen, lateral (soft tissue) neck, cervical spine, or extremities during a 5-month period. A total of 1,471 radiograph examinations was performed, and 200 (14%) misinterpretations (false-positive and false-negative) by the pediatric emergency medicine physicians were identified. As reported previously, 20 (10%) of the false-negative interpretations were noted to be clinically significant, in the current analysis, 103 (7%) false-positive radiograph interpretations were identified. False-positive interpretations were noted more frequently (14%) for soft tissue lateral neck radiographs than for any other radiograph type. Of the 103 total false-positive radiographs, nine (0.6%) resulted in an increased patient cost totaling $764.75. These data show that false-positive radiograph interpretations have limited economic and clinical impact. PMID- 9217524 TI - Child and adolescent suicide attempts: an opportunity for emergency departments to provide injury prevention education. AB - The study objectives were to ascertain whether caretakers of suicidal children and adolescents received emergency department (ED) injury prevention education and to determine if injury prevention education and the medical outcome after a drug overdose are associated with caretakers restricting access to means of suicide. Participants were adult caretakers of children and adolescents who deliberately ingested a drug and received ED evaluation. Information was obtained by poison center chart review and phone interview. Fourteen percent of caretaker reported receiving injury prevention education concerning restriction of access to potential means of suicide at home. ED injury prevention education is significantly associated with caretakers restricting access to suicidal means, even when controlling for medical outcome from the attempt. Because parents are less likely to restrict access to means of suicide without education, injury prevention education about restricting access to means of suicide should be given in the ED. PMID- 9217525 TI - Neural network and linear regression models in residency selection. AB - For many years, multiple linear regression models have been used at a residency program to generate preliminary rank lists of residency applicants. These lists are then used by the admissions committee as an aid in developing a final ranking to submit to the National Residency Match Program (NRMP). A study was undertaken to compare predictions made using linear regression with those generated by a newer technique, an artificial neural network. A prospective cohort design was used. Seventy-four applicants to an emergency medicine program were evaluated by faculty and resident interviewers with regard to medical school grades, autobiography, interviews, letters of recommendation, and National Board scores. Normalization of these scores (by linear transformation of interviewer means) was used to correct for differences among interviewers. Multivariate linear regression and neural network models were developed using data from the previous 5 years' applicants. These models were used to forecast provisional rank orderings of the candidates. These rankings were combined into a single hybrid list that was used by the admissions committee as the starting point for development of the final rank list by consensus. Each model's predictions were tested for goodness of fit against the final NRMP rank using Wilks' test. Using the final submitted NRMP rank order as the dependent variable, the neural network yielded a correlation coefficient of 0.77 and an R2 of 59.4%. The linear regression model exhibited a correlation coefficient of 0.74 and an R2 of 54.0%. No significant difference was found (chi 2 = 1.08, P = .7). A neural network performs as well as a linear regression model when used for forecasting the rank order of residency applicants. PMID- 9217526 TI - The hemolytic uremic syndrome presenting as bilateral leg ischemia. AB - The hemolytic uremic syndrome in adults is an uncommon clinical entity consisting of microangiopathic hemolytic anemia, thrombocytopenia, and renal dysfunction. A previously healthy 42-year-old man, after a 2-day prodromal phase, developed severe pain and coldness in both legs, with purpura in the face and extremities. On admission, hepatorenal dysfunction and disseminated intravascular coagulation were evident. These complicated signs and symptoms led to nonspecific supportive therapy because of delayed diagnosis. The patient's condition gradually improved except for ischemia of the legs, which progressed into symmetrical necrosis; eventually, bilateral below-knee amputation was required. This is the first reported case of the hemolytic uremic syndrome complicated by bilateral leg ischemia. A presumed cause of the ischemia was disseminated intravascular coagulation, a rare complication of the hemolytic uremic syndrome. PMID- 9217527 TI - Complication rates of tube thoracostomy. AB - This study compared the complication rates of tube thoracostomy performed in the emergency department (ED) versus the operating room (OR) and the inpatient ward (IW). A retrospective case series of all patients at an urban, university-based level 1 trauma center hospital who received tube thoracostomy for any indication between 1/1/93 and 12/31/93 was conducted. Complications were defined as empyema, unresolved pneumothorax (persistent air leak or residual pneumothorax), persistent effusion, or incorrect placement. The data for age and duration of tube placement were weighted for analysis of variance (ANOVA). A total of 352 tube thoracostomies was placed in 239 patients. Twenty-three patients had three or more chest tubes placed, 65 had two placed, and the remaining 181 had a single tube. Ninety-nine tubes were placed in the ED, 87 in the OR, and 166 on IW. The mean age of patients in the ED was 37 years, and differed significantly (P < .015) from those in the OR (48 years) and the IW (44 years). The duration of tube placement was similar for all groups (mean = 6.5 days). The overall complication rates related to tube insertion were: ED, 14.0%; OR, 9.2%; IW, 25.3%. Significance was achieved when comparing complication rates between the ED and IW, with less complications in the ED (P = .0436). When comparing complication rates between the ED and OR, there was no significant difference (P = .3643). A power calculation indicated too small of a sample size to truly determine an insignificant difference between complication rates between the ED and OR. Placement of emergent thoracostomy tubes in the ED does not result in an increased complication rate as compared to placement in the IW. PMID- 9217528 TI - Ductus diverticulum interpreted as traumatic aortic injury. AB - A 50-year-old woman was the victim of a motor vehicle accident. An aortogram obtained for suspicion of an aortic injury was interpreted as an intimal disruption at the level of the aortic isthmus. At thoracotomy the patient was found to have only a ductus diverticulum. A brief discussion of the angiographic appearance of a ductus diverticulum and its significance in the setting of trauma is presented. PMID- 9217529 TI - "Refuses to walk" as an ED presentation of acute childhood leukemia. AB - New onset of childhood acute leukemia may present to the emergency department in a variety of ways. This report describes the case of a 3-year-old boy who refused to walk and had minimal physical findings, normal X-rays, and nearly normal lab screening results. His white blood cell count differential led to the diagnosis of acute leukemia. The presentation and evaluation of acute leukemia in children is reviewed. Emergency physicians must be prepared to rule out malignancy in the child who refuses to walk when other more common causes, such as infection and trauma, seem unlikely. PMID- 9217530 TI - Atypical presentation of Henoch-Schoenlein purpura in two children. AB - Henoch-Schoenlein purpura (HSP) is a common vasculitic disorder of childhood. Patients with this disorder typically present with palpable purpura or petechia associated with one or more of the following signs and symptoms: abdominal pain, arthritis/arthralgias, and nephritis. The diagnosis may be difficult to make, however, when a patient presents with isolated symptoms such as abdominal pain without the typical rash. A high index of suspicion must be maintained to diagnose HSP in this setting and to avoid unnecessary interventions. This report describes two unusual patients with the presenting complaint of abdominal pain who had delayed onset of the purpuric rash, making the diagnosis of HSP difficult. PMID- 9217531 TI - Ascertaining hypoglycorrhachia in an acute patient. AB - A study was undertaken in an urgent clinical setting to determine whether the use of a cerebrospinal fluid (CSF) to blood glucose ratio is appropriate for describing the relationships between CSF glucose and blood glucose in patients who had not fasted. Blood glucose levels were obtained before a lumbar puncture in 79 adults who had normal CSF findings. Regression analysis of CSF glucose and blood glucose levels of these patients who had not fasted, as well as data from four published studies of normal blood and CSF glucose levels, indicated that a ratio was not a valid measure of the normal relationship between CSF and blood. Only when the blood glucose level was between 89 and 115 mg/dL was the relationship within the expected "ratio" of 0.60 to 0.70. In hyperglycemic states, the normal relationship may be substantially lower than 0.50. a nomogram is presented which is useful in determining hypoglycorrhachia when the patient is hyperglycemic. PMID- 9217532 TI - Retropharyngeal and epidural abscess from a swallowed fish bone. AB - Retropharyngeal abscess is not uncommon, but the incidence of epidural extension of a retropharyngeal abscess is very rare. Intraspinal involvement of the deep neck infection should be suspected if the patient has neurologic deficits. Emergent surgical drainage and aggressive antibiotic treatment are necessary. The outcome is strongly associated with the level of neurologic function at the time of diagnosis. Contrast-enhanced computed tomography is an excellent diagnostic method for any deep neck infection. A case is presented in which a perforating pharyngeal foreign body (fish bone) induced a retropharyngeal and epidural abscess. The literature is reviewed to improve the early recognition and treatment of this complication of deep neck infection. PMID- 9217533 TI - A difference of 5 degrees C between ear and rectal temperatures in a febrile patient. AB - A 4-year-old boy with a history of seizures triggered by fever presented at an emergency department (ED) with tachycardia, skin vasoconstriction, and a rectal temperature of 42.2 degrees C. However, his ear temperature (as repeatedly measured in two ears, by two experienced nurses, and with two infrared thermometers) was between 36.4 degrees C and 37.6 degrees C. Antipyretic therapy resulted in skin vasodilation, a rapid decrease of rectal temperature, restoration of heart rate, and disappearance of the difference between the two temperatures. Seizures did not occur. This case shows that infrared ear thermometry cannot be recommended in EDs as the procedure of choice for detecting fever in small children, especially when they are vasoconstricted. PMID- 9217534 TI - Traumatic carotid-cavernous sinus fistula with spontaneous resolution. AB - A 77-year-old woman presented with unilateral ocular pain, exophthalmos, vascular tinnitus, and chemosis several weeks after a minor closed head injury. Cerebral angiography showed a carotid-cavernous sinus fistula. One week later the patients's symptoms abruptly ceased. A brief discussion of the presentation and management of these fistulas is presented. PMID- 9217535 TI - Air bag-related ocular trauma. AB - Automobile air bags have recently gained acceptance as an effective measure to reduce the morbidity and mortality associated with motor vehicle accidents. This report describes 11 cases of air bag-related ocular trauma and reviews cases previously reported in the literature, for a total of 32 patients and 39 eyes. This is the first comprehensive report on various types of ocular trauma related directly to air bag deployment. The most common type of ocular injuries seen are to the eyelids (23 patients, 28 eyes), conjunctiva (21 patients, 25 eyes), and cornea (23 patients, 28 eyes). Hyphema was frequently seen (10 patients, 11 eyes). Several serious cases of vision-threatening injuries, including retinal detachment, retinal dialysis, scleral rupture, and dislocated lens, were also reported. The following patterns were found: 55% of patients were male and 45% female; ages ranged from 2 to 81 years with a mean age of 36 years; the right eye was involved in 35% of cases, the left in 38%, and 27% were bilateral. Based on these findings, it is recommended that all patients who present with air bag related ocular trauma undergo a complete ophthalmologic examination because the high-velocity blunt trauma causes ocular injuries that may be more serious than they initially appear. Further refinements in design and deployment need to be made to reduce the frequency and severity of air bag-related ocular injuries. PMID- 9217536 TI - Creating a home page on the World Wide Web: an inexpensive means to promote medical education and physician recruitment. AB - The World Wide Web (WWW) is generally used as an information resource. It can also be used as a national and international promotional (advertising) resource, at minimal cost, to assist in physician recruitment, such as for residency training programs. Currently, only a few residency training programs have home page sites describing their programs on the WWW. Creating a WWW site that can be viewed nationally and internationally requires an internet service provider and hypertext markup language (HTML) document that designs the WWW home page site. This article provides a step-by-step method for creating a simple WWW site (including an HTML template) to promote a residency program and assist in resident recruitment. As more young physicians graduate with more extensive computer skills and familiarity, use of the WWW for physician recruitment will become a more important source of information for physician applicants. PMID- 9217537 TI - Bedside diagnostic testing of body fluids. AB - Numerous bedside diagnostic modalities are appropriate for the practice of emergency medicine. The proliferation of sophisticated technology is likely to increase both the availability and accuracy of commercial testing products. If health care reform in the United States results in a relaxation of the CLIA regulations, there will be a rapid expansion of research and development aimed at the biotechnology market. How much this would pertain to hospital-based emergency practice remains to be seen. Cost containment pressures may act in both directions on the utilization of available bedside technology. Although these tests are often less expensive than centralized laboratory determinations, the ready availability of near-patient testing may result in an increase in use that negates the lower cost. As with other diagnostic modalities, a thoughtful, considered approach based on scientific evidence will be necessary to formulate the appropriate use of bedside testing in individual emergency practice settings. PMID- 9217538 TI - Dorsal dislocation of the four ulnar metacarpals. AB - A patient presented to the emergency department (ED) with hand pain after striking a wall with a partially clenched fist. X-rays showed small dorsal carpal avulsion fractures and a dorsal dislocation of the ulnar four metacarpals. Closed reduction maneuvers reduced all but the second metacarpal. The patient underwent open reduction, internal fixation of this joint, and was healed within 6 weeks. Although typically associated with significant amounts of kinetic energy, the particular position of the hand and wrist during this injury may have allowed this dislocation pattern to occur with relatively minor trauma. A discussion of this injury, the carpometacarpal joint, and important radiographic features is presented. PMID- 9217539 TI - Isolated talus fractures: description of a new clinical sign. AB - Isolated talus fractures are very uncommon and are usually associated with severe trauma. Five cases of isolated talus fracture associated with relatively minor trauma are reported. These five patients had one clinical sign in common, pain out of proportion to the severity of their injury. The treatment of talus fractures is reviewed. PMID- 9217540 TI - Cases in electrocardiography. PMID- 9217541 TI - Dilemmas in the acute pharmacologic treatment of uncontrolled atrial fibrillation. AB - A recently conducted observational study of the prehospital treatment of uncontrolled atrial fibrillation brought to light therapeutic inconsistencies by emergency providers in dealing with this dysrhythmia. A review of the literature suggests that digoxin lacks efficacy in controlling ventricular rate in atrial fibrillation and that the slow onset of digoxin makes its use in the emergency setting questionable. Because of their demonstrated ability to rapidly slow ventricular rate, the calcium channel blocker, diltiazem, or the beta-adrenergic blocker, esmolol, should be the preferred agents for treating rapid atrial fibrillation in the emergency department or the paramedic ambulance. PMID- 9217542 TI - Medical-legal considerations related to symptom duration and patient outcome after bacterial meningitis. PMID- 9217543 TI - Emergency medicine in Cuba: report from a country in isolation. AB - Cuba is one of the poorest countries of the world. For the past 34 years the United States has maintained an economic embargo against Cuba. Because of this, Cuba has not been able to share in advances in American medical technology, pharmaceuticals, or research. The country has also been abandoned by the former Soviet Union. Recent political developments have assured continuation of the embargo. This report describes the current state of prehospital and emergency medical care in Cuba, how the embargo has affected emergency services, and possibilities for the future. PMID- 9217544 TI - Critical care medicine: an annotated bibliography of the recent literature. PMID- 9217545 TI - Research directions in emergency medicine. PMID- 9217546 TI - Transient apnea with intramuscular ketamine. PMID- 9217547 TI - Totally implantable vascular access and emergent management of refractory recurrent grand mal status epilepticus. PMID- 9217548 TI - Hypomagnesemia. PMID- 9217549 TI - Caustic instillation into the ear as a form of child abuse. PMID- 9217550 TI - Spontaneous rupture of the spleen in amyloidosis. PMID- 9217551 TI - A new phenytoin prodrug, Cerebyx. PMID- 9217552 TI - Ruptured abdominal aortic aneurysm masquerading as testicular pain. PMID- 9217553 TI - Office evaluation of the patient with musculoskeletal complaints. AB - Many musculoskeletal complaints are accompanied by classic signs and symptoms that can be readily diagnosed by the primary care physician. Others are much less obvious and present a diagnostic challenge. In the office evaluation of patients with musculoskeletal complaints, the history is the most informative element. Least helpful are laboratory tests. Although erythrocyte sedimentation rate (ESR), rheumatoid factor, and other widely available tests are sensitive to the presence of rheumatic diseases, they are not specific for any of them. In the initial office evaluation, helpful points of differentiation include the number of joints involved, their location, and, when multiple joints are involved, whether they are symmetric or asymmetric. An acute monarthritis is associated mainly with trauma, infection, or a crystal-induced synovitis such as gout or pseudogout. Patients with polyarthritis may have symptoms that come and go very quickly, sometimes in < 24-36 hours. This migratory pattern characterizes diseases such as gonococcal arthritis, viral disease, and sarcoidosis. "Rheumatoid variants" such as Reiter's syndrome, psoriatic arthritis, and spondylitis may affect no more than a few joints and are accompanied by other signs, such as nail and skin lesions (psoriasis) or urogenital and enteric infections (Reiter's). Like erosive osteoarthritis, the rheumatoid variants may also cause swelling and inflammation of the distal interphalangeal joints. The classic example of symmetric joint disease is rheumatoid arthritis (RA). While RA often occurs in a progressive and additive pattern, its onset may be followed by a remission several months later. Patients who present with the "algias" may have no physical signs but manifest extensive musculoskeletal pain. Fibromyalgia occurs typically in younger women; polymyalgia rheumatica rarely occurs in patients < 50 years of age and is usually accompanied by a strikingly high ESR. Age and gender should be noted in the office evaluation because they can provide clues not only to these "algias," but other rheumatic diseases seen more frequently in one age or gender group than another. PMID- 9217555 TI - A clinician's approach to acute low back pain. AB - Two important goals in treating acute low back pain are to return the patient to regular activity as quickly as possible and to do so in a manner that is cost effective. By following a logical treatment protocol, the clinician is often able to provide the treatment necessary to provide the patient with relief. Referral to an orthopedist or neurosurgeon may be appropriate in only a minority of cases. Thus, after the initial history and physical examination, ruling out (or in) conditions that require urgent or emergent care is essential. These conditions include cauda equina syndrome, circulatory collapse due to expanding abdominal aortic aneurysm, and tumor, infection, and other underlying disorders as a cause of low back pain. Patients without these conditions can be started on conservative therapy-without radiographic or laboratory tests-regardless of the specific diagnosis. Conservative therapy consists of passage of time, controlled physical activity, physical modalities (e.g., cryotherapy or thermotherapy), local injections, nonsteroidal anti-inflammatory drugs, and muscle relaxants. Because low back pain is so common, even the small proportion of patients who do not improve after 6 weeks of conservative therapy represents a sizable number. The location and radiation of pain are used as initial guides to classifying these patients into four groups: those with localized pain, sciatica, anterior thigh pain, or posterior thigh pain. Each follows a different diagnostic path, which will be described herein. PMID- 9217554 TI - Emerging treatments for rheumatoid arthritis. AB - Rheumatoid arthritis was considered for centuries to be a nuisance condition, limiting in its effects on an individual's range of motion and the source of considerable distress, but not a life-threatening disease. Recently, however, it has become apparent that patients with severe rheumatoid arthritis may have a decreased life span. Current pharmacologic therapies for patients with rheumatoid arthritis, which include nonsteroidal anti-inflammatory drugs, disease-modifying antirheumatic drugs, methotrexate, and corticosteroids, have been moderately successful in alleviating the discomforts associated with swollen, painful joints. Many practitioners have sought to improve use of these agents and slow joint destruction by challenging traditional treatment paradigms, altering the sequence in which drugs are given. Nevertheless, most standard medical approaches to treatment have had little or no impact on the course of rheumatoid disease. Innovative strategies, particularly those based on new concepts in the immunobiology of rheumatoid arthritis, are being developed to target cellular inflammatory mechanisms and actually prevent disease progression. Some agents, such as inhibitors of 5-lipoxygenase-omega-3 fatty acid and zileuton-may be most useful in treatment of milder disease manifestations such as moderate synovitis. Other agents, such as oral type II collagen, minocycline, subcutaneous interleukin-1ra, and anti-CD4 monoclonal antibodies, have produced such inconsistent results that substantial additional research will be required before any conclusions may be drawn about their value. Among the most promising agents, and the most extensively studied, are tumor necrosis factor-alpha monoclonal antibodies, immunosuppressive drugs such as cyclosporine and mycophenolate mofetil, and the novel compound tenidap, which has both cytokine-modulating and anti-inflammatory properties. PMID- 9217556 TI - Soft-tissue rheumatism: diagnosis and treatment. AB - Soft tissue rheumatism is one of the most common and most misunderstood categories of disorders facing the primary care physician. Among the more common types are subacromial bursitis, epicondylitis, trochanteric bursitis, anserine bursitis, and fibromyalgia. The keys to the diagnosis of soft-tissue rheumatism are the history and, more importantly, the physical examination. Extensive laboratory testing and radiographs are not as helpful in evaluating patients with these complaints. Treatment consists of nonsteroidal anti-inflammatory drugs (NSAIDs) and nonnarcotic analgesics. Especially in patients with localized disorders, intralesional injections of corticosteroids are particularly effective and safe and should be part of the armamentarium of the primary care practitioner. Fibromyalgia is a particularly challenging form of nonarticular rheumatism. The clinical presentation is rather characteristic, with the patient typically being a woman 30-60 years of age who presents with diffuse somatic pain. Patients often give a history of sleep disturbance, may be depressed, and show characteristic tender areas, or trigger points. Laboratory findings are normal. Management includes reassurance, correction of the underlying sleep disturbance with low doses of a tricyclic antidepressant, treatment with muscle relaxants and nonnarcotic analgesics or NSAIDs, and an exercise program with a strong aerobic component. PMID- 9217559 TI - Liability issues in the treatment of patients with rheumatic diseases. AB - In clinical practice, risk management focuses primarily on concerns regarding adverse occurrences and the possibility of resulting civil litigation. Tort law, one of the major branches of civil law, is most relevant to clinical risk management. Duty, breach, causation, and damages are the four elements of the tort of professional liability, i.e., medical malpractice. Liability concerns in clinical practice commonly involve adverse drug reactions. Other potential liability concerns include informed consent, adverse outcomes, and economic considerations. PMID- 9217557 TI - Monarthritis: differential diagnosis. AB - Acute monarthritis should be regarded as infectious until proved otherwise. Early evaluation is crucial because of the capacity of some infectious agents to destroy cartilage rapidly. The history and physical examination can provide highly suggestive clues, but a definitive diagnosis may depend on arthrocentesis and analysis of synovial fluid. The diagnosis of acute monarthritis is rarely established by radiography. The most common cause of bacterial arthritis is Neisseria gonorrhoeae. Staphylococcus aureus and streptococci are the organisms most frequently implicated in nongonococcal bacterial arthritis, although the possibility of Gram-negative bacteria or anaerobes should not be overlooked in intravenous drug users or immunocompromised patients. Inflammation in a large joint, particularly the knee, might arouse suspicion of Lyme disease. Other, less frequently encountered infectious causes of acute monarthritis include tuberculosis and other mycobacteria, fungi, and viruses. Arthroscopic examination and synovial tissue biopsy may be necessary to diagnose such processes. Microscopic examination of the synovial fluid may reveal a crystalline etiology for monarthritis. Monosodium urate crystals induce gout, usually in the toe, ankle, or midfoot, while calcium pyrophosphate crystals cause pseudogout, most often in the knee or wrist. Acute monarthritis is sometimes a manifestation of osteoarthritis or an early sign of a systemic arthritis such as rheumatoid or reactive arthritis. Processes underlying acute monarthritis can also evolve into a more chronic clinical picture as exemplified by the spondyloarthropathies. PMID- 9217558 TI - Osteoporosis: prevention, diagnosis, and management. AB - Osteoporosis is a public health scourge that is usually eminently preventable. Some risk factors, such as low calcium intake, vitamin D deficiency, and physical inactivity, are amenable to early interventions that will help maximize peak bone density. Other risk factors subject to modification are cigarette smoking and excessive consumption of protein, caffeine, and alcohol. Hip fractures are the most serious outcome of osteoporosis, with enormous personal and public health consequences. The ongoing Study of Osteoporotic Fractures has identified additional independent predictors of hip fracture risk, including maternal hip fracture, absence of significant weight gain since age 25, height, hyperthyroidism, use of long-acting benzodiazepines or anticonvulsants, spending < 4 hours a day on one's feet, inability to rise from a chair without using one's arms, poor visual depth perception and contrast sensitivity, and tachycardia. In an individual perimenopausal woman, the risk of osteoporotic fracture and the urgency of estrogen replacement therapy can be best estimated on the basis of bone mineral density, as measured by dual-energy x-ray absorptiometry, coupled with the presence or absence of existing fractures and clinical risk factors evident from the history and physical examination. Estrogen, calcitonin, and bisphosphonates have all been proved effective in retarding postmenopausal bone loss and therefore reducing the risk of fracture. The use of sodium fluoride is more controversial, although a recent study has suggested a possible role for slow-release fluoride combined with high-dose calcium supplementation. PMID- 9217560 TI - Creating solutions to meet the challenges facing biomedical research: report of the 1996 APM Fall Symposium. PMID- 9217561 TI - How long, oh how long? PMID- 9217562 TI - The opinion of current and recent internal medicine residents regarding a fourth year of training and the future of general internal medicine. American College of Physicians National Council of Associates. AB - OBJECTIVE: To determine the opinion of current residents and recent graduates of internal medicine training programs regarding an additional mandatory year of residency training. METHODS: A survey was made of 2,000 associate members of the American College of Physicians from five geographic regions. RESULTS: Of 917 respondents, 70.3% thought a fourth year of training would impact negatively on their choice of a career in internal medicine, and 82.9% believed a mandatory fourth year should not be required of residents choosing a subspecialty career. Furthermore, 58.1% of physicians surveyed thought a mandatory fourth year would discourage individuals from pursuing subspecialty careers. If a mandatory fourth year of training were required, 50.7% respondents indicated that it should consist of ambulatory training in a number of general fields, while 49.6% physicians believed the training should focus on one or two subspecialty fields. CONCLUSIONS: A mandatory fourth year of training is not supported by residents and recent graduates of the programs surveyed. PMID- 9217563 TI - Calcium antagonists in cardiovascular disease: a necessary controversy but an unnecessary panic. PMID- 9217564 TI - Myocardial infarction in newly diagnosed hypertensive Medicaid patients free of coronary heart disease and treated with calcium channel blockers. AB - PURPOSE: A retrospective cohort analysis of 1,406 newly diagnosed hypertensive patients, aged 18 to 59, without prior coronary heart disease and initially treated with calcium channel blockers (CCBs) or eight other drug regimens was conducted to evaluate the relative risk of acute myocardial infarction (AMI) among patients on CCBs alone or with a diuretic. MATERIALS AND METHODS: Administrative claims data from Pennsylvania's Medicaid program were the data source. Patients were followed up from 1987 to 1994. RESULTS: There was a highly significant trend towards prescribing CCBs between 1988 and 1991 (P = 0.0001). A total of 67 AMIs occurred, 33 of which were during original drug therapy. Compared with those treated with beta blockers, the relative risk of AMI among patients treated only with CCBs was 0.49 (95%) confidence interval [CI] 0.11 to 2.20). Compared with diuretic therapy, the AMI relative risk associated with CCB therapy was 0.60 (95% CI 0.16 to 2.32) when patient drug regimen was classified based on the first six prescriptions. Several alternative drug regimen classification schemes were tested to evaluate the sensitivity of relative risk of AMI to classification of drug therapy. CONCLUSIONS: The measurement of relative risk of AMI was highly dependent on the study design, including patient selection criteria and classification of patient drug therapy. Previous findings of elevated risk of AMI from CCB antihypertensive therapy could not be confirmed. PMID- 9217565 TI - Predictors of coronary artery disease in patients with cocaine-associated myocardial infarction. Cocaine-Associated Myocardial Infarction (CAMI) Study Group. AB - PURPOSE: To identify clinical criteria predictive of underlying coronary artery disease in patients with cocaine-associated myocardial infarction. PATIENTS AND METHODS: Using a retrospective cross-sectional study design at 29 acute care hospitals, we identified 70 patients with cocaine-associated myocardial infarction who had a determination of the presence or absence of coronary artery disease. Clinical characteristics of patients with coronary artery disease (> 50% stenosis on cardiac catheterization or reversible ischemia on stress test) were compared with patients without coronary artery disease (< 50% stenosis on cardiac catheterization). RESULTS: Compared with patients without coronary artery disease (n = 21), patients with coronary artery disease (n = 49) were older (42 versus 31 years; P < 0.001), had more traditional cardiac risk factors (2.3 versus 1.5; P < 0.001), more frequent history of hypertension (odds ratio [OR], 5.3; 95% confidence interval [CI], 1.4 to 20.4); more frequent family history of myocardial infarction (OR, 4.4; 95% CI, 1.3 to 15.1), more bradydysrhythmias (OR, 8.0; 95% CI, 1.0 to 65.5), and more likely to have an inferior infarct location (P = 0.04). CONCLUSION: Age, number of cardiac risk factors, location of myocardial infarction, and bradydysrhythmias predict underlying coronary artery disease in patients with cocaine-associated myocardial infarction. If validated, this knowledge may be used to develop a medically appropriate, cost-effective evaluation strategy for patients following cocaine-associated myocardial infarction. PMID- 9217566 TI - Nonadherence in tuberculosis treatment: predictors and consequences in New York City. AB - BACKGROUND: Poor adherence to antituberculosis treatment is the most important obstacle to tuberculosis control. PURPOSE: To identify and analyze predictors and consequences of nonadherence to antituberculosis treatment. PATIENTS AND METHODS: Retrospective study of a citywide cohort of 184 patients with tuberculosis in New York City, newly diagnosed by culture in April 1991-before the strengthening of its control program-and followed up through 1994. Follow-up information was collected through the New York City tuberculosis registry. Nonadherence was defined as treatment default for at least 2 months. RESULTS: Eighty-eight of the 184 (48%) patients were nonadherent. Greater nonadherence was noted among blacks (unadjusted relative risk [RR] 3.0, 95% confidence interval [CI] 1.1 to 8.6, compared with whites), injection drug users (RR 1.5, 95% CI 1.1 to 2.0), homeless (RR 1.4, 95% CI 1.0 to 1.8), alcoholics (RR 1.4, 95% CI 1.0 to 1.9), and HIV infected patients (RR 1.4, 95% CI 1.1 to 1.9); also, census-derived estimates of household income were lower among nonadherent patients (P = 0.018). In multivariate analysis, only injection drug use and homelessness predicted nonadherence, yet 46 (39%) of 117 patients who were neither homeless nor drug users were nonadherent. Nonadherent patients took longer to convert to negative culture (254 versus 64 days, P < 0.001), were more likely to acquire drug resistance (RR 5.6, 95% CI 0.7 to 44.2), required longer treatment regimens (560 versus 324 days, P < 0.0001), and were less likely to complete treatment (RR 0.5, 95% CI 0.4 to 0.7). There was no association between treatment adherence and all cause mortality. CONCLUSIONS: In the absence of public health intervention, half the patients defaulted treatment for 2 months or longer. Although common among the homeless and injection drug users, the problem occurred frequently and unpredictably in other patients. Nonadherence may contribute to the spread of tuberculosis and the emergence of drug resistance, and may increase the cost of treatment. These data lend support to directly observed therapy in tuberculosis. PMID- 9217567 TI - Natural history of blood pressure in sickle cell disease: risks for stroke and death associated with relative hypertension in sickle cell anemia. AB - PURPOSE: Blood pressure in individuals who have sickle cell disease has been reported to be lower than published normal values. We determine whether and to what degree this is true, using data obtained as part of a large natural history study. PATIENTS AND METHODS: Blood pressure was measured annually for 3,317 subjects with sickle cell disease who were 2 years old or older. Values obtained were compared with those reported by the National Health and Nutrition Examination Survey I and II (NHANES I and II). They were further analyzed with respect to age, sex, height, weight, hematologic diagnosis, blood urea nitrogen and creatinine, stroke, and death. RESULTS: Blood pressure was significantly lower in subjects with sickle cell anemia than published norms for age, race, and sex, a difference that increased with age. It correlated with body mass index, hemoglobin, measures of renal function and age, but the strength of the correlation varied among age and sex subgroups. The risk for occlusive stroke increased with systolic but not diastolic pressure. Mortality was related to elevated blood pressure in males (P < 0.05) and to a lesser extent in females (P = 0.10). In subjects with hemoglobin SC disease, blood pressure also deviated from normal but to a lesser degree. CONCLUSION: Blood pressure is generally lower than normal in individuals with sickle cell anemia. Those with high values relative to this population had an increased risk of stroke and death. Blood pressure should be monitored but values obtained must be assessed relative to the lower values expected for patients with this disease. Those with blood pressure values above 140/90 mm Hg should be evaluated and considered for treatment. PMID- 9217568 TI - Antiendothelial cell antibodies: useful markers of systemic sclerosis. AB - BACKGROUND: Systemic sclerosis (SS) encompasses a wide spectrum of clinical presentations. Antiendothelial cell antibodies (AECA) in patients with primary Raynaud's phenomenon (PRP), limited SS (lSSc), or diffuse SS (dSSc) may help to determine the long-term prognosis of the disease. METHODS: Twenty-seven normal controls, 13 patients with PRP, 36 with lSSc, and 31 with dSSc were included in the study. Sera were examined for the presence of AECA, using a cellular enzyme linked immunosorbent assay (ELISA). Angiotensin-converting enzyme (ACE) activity, plasma von Willebrand factor antigen (vWfAg), and thrombomodulin (Tm) concentrations were also evaluated. The medical records of 50 of the lSSc and dSSc patients were reviewed and the organ system involvement noted. RESULTS: Antiendothelial cell antibodies were present in 3 patients with PRP, 16 patients with lSSc, and 26 patients with dSSc. These autoantibodies were mainly of the IgG isotype. There was no difference in ACE activity between patients and controls. In contrast, vWfAg and Tm concentrations were higher in patients with PRP relative to controls, and higher in patients with lSSc compared with those with PRP. The presence of AECA was associated with digital scars and ulcers (P < 0.004 and P < 0.003, respectively), severe RP (P < 0.01), grade 3 tortuosity of vessels (P < 0.0004), and lung involvement (P < 0.02). CONCLUSION: The significant trend for AECA to increase with disease severity across the three groups of patients studies suggests that the AECA test can identify subsets of SSc with differing prognoses. PMID- 9217569 TI - Clinical features and outcome of 95 patients with hypersensitivity vasculitis. AB - PURPOSE: To evaluate the clinical features and outcome of patients with isolated hypersensitivity vasculitis (HV). PATIENTS AND METHODS: Retrospective study of patients with cutaneous vasculitis followed up at a University Hospital from 1975 to 1994. Patients with vasculitis secondary to collagen vascular diseases, neoplasia, or major infections were excluded. Patients were classified as HV according to the differential criteria proposed by Michel et al (J Rheumatol. 1992;19:721-728). RESULTS: Ninety-five patients were classified as HV. The mean age was 42.7 +/- 21.7 years, with similar disease frequency in both sexes. In 43 patients, the precipitating event was drug therapy, either alone or as a treatment for a coexistent infection, usually an upper respiratory tract infection. The most frequent clinical manifestation was palpable purpura followed by joint symptoms. Systemic involvement was infrequent: 7 patients had nephropathy, manifested almost exclusively by microhematuria, and 5 patients had gastrointestinal symptoms. In 54 subjects the vasculitis did not require treatment; 26 patients were treated with NSAIDs, and 14 required corticosteroids (associated to immunosuppressive agents in 2 of them). After a mean follow-up of 15.5 +/- 28.9 months (median 6), only 2 patients had slight renal impairment, whereas the remaining had a complete recovery. CONCLUSION: Hypersensitivity vasculitis is usually a benign syndrome, often secondary to drugs or infections, or both. Its main clinical manifestations are skin and joint symptoms. The systemic involvement is scarce and its prognosis is excellent. PMID- 9217570 TI - Analysis of long-term endoscopic surveillance during follow-up after variceal sclerotherapy from a 13-year experience. AB - PURPOSE: To evaluate the course of patients with bleeding esophageal varices treated with endoscopic sclerotherapy after obliterating varices and to determine the cost benefits of long-term endoscopic surveillance from a retrospective analysis of a 13-year experience. LOCATION: University-affiliated teaching hospital and county facility. METHODS: Patients whose varices were obliterated by endoscopic sclerotherapy were considered for the study if they had a minimum of 12 months of follow-up. Sclerotherapy was initially performed weekly, increasing intervals to eventual yearly treatments. Varices were reobliterated if they reformed. Variables assessed were rebleeding, mortality, employment status, and cost based on allowable and reimbursed Medicare rates. RESULTS: Of 324 patients who achieved variceal obliteration, analysis included 104 eligible patients who were followed up for > 12 months (41 +/- 28). Varices reformed in 73 patients (71%), mostly in the first year after obliteration or reobliteration. Abstinent alcoholic patients were least likely to reform varices. Nineteen patients (18%) had 23 rebleeding episodes, and in 10 patients (10%) portalsystemic shunt was placed. Survival was 84% and bleeding-related mortality was 6%. Significantly more patients were employed while on the program compared with entry. The yearly cost of treating variceal reformers ($2,117) was significantly higher than variceal nonreformers ($1,735), but the overall cost of maintaining a patient on a chronic sclerotherapy program was relatively small. CONCLUSIONS: The low rebleeding, low mortality, and relatively low cost in patients managed long term by chronic sclerotherapy underscores the benefits of this treatment program. PMID- 9217571 TI - Pathophysiology of Helicobacter pylori-induced gastritis and peptic ulcer disease. AB - Helicobacter pylori causes persistent infection and inflammation in the human stomach, yet only a small fraction of persons harboring this organism develop peptic ulcer disease. An important question is why this variation in infection outcome exists. Recent studies have demonstrated that H pylori isolates possess substantial phenotypic and genotypic diversity that may engender differential host inflammatory responses that influence clinical outcome. Further investigation in this field may help to define which H pylori-infected persons bear the highest risk for subsequent development of peptic ulcer disease, and thus enable physicians to focus eradication therapy. PMID- 9217572 TI - Toxicity of nonsteroidal anti-inflammatory drugs in the elderly: is advanced age a risk factor? AB - We reviewed the pharmacokinetic, physiologic and epidemiologic data concerning nonsteroidal anti-inflammatory drug (NSAID)-induced gastropathy and renal insufficiency in the elderly through a structured critical reading of the literature. References were collected through a search of MEDLINE and consultation with experts in the field. While there is an abundance of pharmacokinetic data comparing relevant parameters in young and old subjects, methods are not uniform and findings are inconsistent. Prostaglandin physiology appears to be altered in older versus younger subjects. Most surprisingly, there is a scarcity of epidemiologic data examining the contribution of age as a risk factor for NSAID-induced ulcers and/or renal insufficiency. The data that do exist do not clearly support age as an independent risk factor; and we believe that comorbidities, comedications and past history are more important predictors of NSAID-induced toxicity than age and more relevant in regard to therapeutic decision-making for this patient population. PMID- 9217573 TI - Leukocytoclastic vasculitis associated with nizatidine therapy. PMID- 9217574 TI - Azithromycin-induced intrahepatic cholestasis. PMID- 9217575 TI - Hepatitis C-associated osteosclerosis after blood transfusion. PMID- 9217576 TI - Iatrogenic ferroma: a cause of failure of serum ferritin to respond to phlebotomy. PMID- 9217577 TI - Prophylaxis of visceral leishmaniasis in human immunodeficiency virus-infected patients. PMID- 9217578 TI - Epidemiology and treatment of hypercholesterolemia: where we are today. Introduction. PMID- 9217579 TI - Hypercholesterolemia in the United States: how far have we come? AB - In a series of clinical trials and epidemiologic studies over the past 30 years, clear associations have been established between saturated fat in the diet, elevated serum cholesterol levels, and the risk of coronary heart disease (CHD). This understanding has been applied in intervention trials using diet, exercise, and drugs to lower cholesterol. The National Cholesterol Education Program was established in 1985 to coordinate efforts to make U.S. citizens aware of the risks of elevated cholesterol and to provide guidelines for lowering it. Trials over the last several years have demonstrated benefit of cholesterol lowering in secondary prevention and producing regression of coronary lesions. While much progress has been made, we are still far from the goal of eliminating hypercholesterolemia in the United States. Improved educational efforts and new strategies for cholesterol management are needed. PMID- 9217580 TI - Nonlipoprotein risk factors for coronary heart disease: evaluation and management. AB - Appropriate prevention and management of coronary heart disease (CHD) requires an integrated approach to the reduction of risk factors. These principally include reduction of elevated lipids, control of blood pressure, and cessation of smoking. In addition, appropriate exercise, diet, and weight reduction (where necessary) are also important. Control of diabetes and stress management may also be helpful. Aspirin therapy is appropriate for all patients with known CHD and selected patients without CHD who have several risk factors, including nonmodifiable risk factors such as age, a positive family history, and male gender. PMID- 9217581 TI - Management of high serum cholesterol and related disorders in patients at risk for coronary heart disease. AB - Cholesterol lowering has been shown to be of benefit in reducing the risk of coronary heart disease (CHD) in both patients with established CHD (secondary prevention) and those without (primary prevention). In secondary prevention trials, moderate cholesterol lowering reduced the rate of new events and decreased both morbidity and mortality from cardiovascular disease. In primary prevention, a reduction of cholesterol by 20% has produced a 31% reduction in recurrent coronary morbidity, a 33% reduction in coronary mortality, and 22% less total mortality. The target of therapy is low-density lipoprotein (LDL) cholesterol: in patients with established CHD, goal LDL is < or = 100 mg/dL. In high-risk patients without established CHD, the target goal for LDL cholesterol is < or = 130 mg/dL. Nondrug measures, bile acid sequestrants, nicotinic acid, and 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors all play important roles in cholesterol-lowering therapy. PMID- 9217582 TI - A population-based approach to cholesterol control. AB - The large proportion of the population who have only modest or moderate hypercholesterolemia will experience more coronary events than the smaller percentage of people who are at higher risk from more extreme elevations of serum cholesterol. The high-risk individual strategy for prevention of coronary heart disease (CHD) can result in impressive declines in cholesterol, but the benefits will be concentrated at the upper end of the population distribution. On the other hand, a population strategy for coronary disease prevention will achieve a much more modest reduction of cholesterol, but these changes will be over the entire distribution and will reduce the risk of the much larger proportion of people with average cholesterol levels who otherwise would likely go untreated. In light of the fact that epidemiologic studies and long-term clinical trials predict that a 10% reduction in serum cholesterol will result in a 30% reduction in coronary events, the population strategy has the potential for an enormous impact in reducing CHD, which, despite our great success over the past two decades, still remains the number one killer in the United States. PMID- 9217583 TI - Cholesterol screening should be targeted. AB - For primary prevention of coronary heart disease (CHD), the American College of Physicians (ACP) has recommended that initial cholesterol screening be targeted to people who have other risk factors in addition to elevated cholesterol. This would include those with symptoms of heart disease, asymptomatic men 35-65 years old and women 45-65 years old, or younger people who have > or = 2 risk factors or who might benefit from treatment for high blood cholesterol. After the age of 75, cholesterol is no longer a risk factor, so there is no rationale for testing. In primary prevention, lipoprotein fractionation should be performed in men and women who have been identified as having elevated blood cholesterol levels, not as part of initial testing. In secondary prevention, some studies indicate that cholesterol reduction may be beneficial after age 65. In asymptomatic younger people without other risk factors, the low prevalence of CHD and rapid response to cholesterol reduction once it is initiated suggest that early screening and treatment are unnecessary. Everyone should adopt the lifestyle modifications conducive to cardiovascular health, but the ACP believes that, for primary prevention, universal screening is neither cost effective nor the best use of the patient's and physician's time. PMID- 9217584 TI - Adults aged 20 and older should have their cholesterol measured. AB - The guidelines of the National Cholesterol Education Program recommend that adults > or = 20 years of age should have their total and high-density lipoprotein cholesterol measured. This recommendation, which has been endorsed by representatives of > 40 medical and health organizations, is based on a large and diverse body of scientific evidence derived from animal, pathologic, genetic, biochemical, metabolic, and epidemiologic studies and clinical trials. Elevated cholesterol levels raise the risk of coronary heart disease (CHD) in men and women and in younger and older adults. Recent clinical trials have confirmed that cholesterol lowering reduces CHD morbidity and mortality and total mortality, without an increase in noncardiovascular mortality, in patients with and without CHD. Measuring cholesterol levels in adults > or = 20 years of age is necessary to provide an accurate assessment of CHD risk to an individual; to identify individuals who should lower their cholesterol levels, using diet and lifestyle changes as the primary treatment; and to reinforce population recommendations. Atherosclerosis begins early in life, and cholesterol levels in young adults predict CHD risk 30-40 years later. Cholesterol measurement can be used to motivate lifestyle changes that will reduce the long-term risk for CHD. Waiting until mid-life to find an elevated cholesterol loses a significant portion of the benefit. Cholesterol is a CHD risk factor in women and older adults, and recent trials show significant CHD risk reduction in these groups. While drug treatment is properly directed to patients with high CHD risk, in whom drugs are cost effective, cholesterol measurement and lifestyle-based cholesterol lowering are necessary on a broader scale to reduce long-term CHD risk in adults aged > or = 20 years. PMID- 9217585 TI - Current status of cholesterol treatment in the community: the Minnesota Heart Survey. AB - There is a consensus on the importance of lowering blood cholesterol in individuals and populations. To determine trends in the detection and treatment of elevated cholesterol, a series of studies known as the Minnesota Heart Survey evaluated cardiovascular disease, risk, and health behavior among adults in the upper Midwest between 1980 and 1992. Over 25,000 adult residents of large and small communities were surveyed for information on risk factors and health habits, including status of cholesterol detection and treatment. During those years, population levels of blood cholesterol declined significantly for both men and women, largely as the result of changes in diet. Levels of clinical detection of hypercholesterolemia, initially low, also rose. However, subjects who had been informed that they had increased lipids reported that recommendations from their physicians for dietary therapy declined, while recommendations for weight loss increased during the survey period. Medication use for elevated blood cholesterol, always low, rose slightly, but many subjects discontinued medications due to side effects, the perception that their cholesterol was controlled, lack of perceived benefit, or cost. A total of 274 primary care physicians were also surveyed. Physicians reported that they screen more frequently than in the past and initiate drug therapy at a lower threshold. Despite improving trends in detection, treatment, and follow-up for elevated blood cholesterol in the general population, > 50% of U.S. citizens are still unaware of their elevated cholesterol levels and a growing segment of the population that has been identified as having elevated blood cholesterol remains untreated. Dietary therapy needs to be better utilized. Physicians also need to educate their patients about the importance of maintaining desirable cholesterol levels and to encourage compliance with medications for those who require them. PMID- 9217586 TI - Compliance with treatment regimens in chronic asymptomatic diseases. AB - At least one third of hospital admissions for heart failure result from noncompliance with therapeutic regimens, both dietary and pharmacologic. In chronic diseases, noncompliance with both lifestyle modification and medication regimens is a major health problem. Patients frequently stop taking their medications because they consider them ineffective or because they experience unpleasant side effects. In asymptomatic conditions, patients may believe they do not need the medication and may not even fill their prescription. If they do obtain the medications, they may forget to take them regularly. Educational efforts and behavioral techniques can improve patient compliance in chronic, asymptomatic conditions, but one of the most effective strategies remains improved patient-physician communication. PMID- 9217587 TI - Healthcare decisions and product labeling: results of a consumer comprehension study of prototype labeling for proposed over-the-counter cholestyramine. AB - This study was designed to determine whether randomly selected subjects could comprehend prototype consumer-oriented package labeling and inserts for over-the counter cholestyramine, a nonsystemic lipid-lowering agent. The primary messages communicated in the label were that consumers should see their doctor before taking cholestyramine and should read the full package insert. In addition, the label communicated indication, dosage, and preparation, as well as key warnings about use with other medications. The insert reinforced the message about seeing the doctor before taking cholestyramine and before taking concurrent medications, further explained the purpose of the drug and its correct use, and provided information about the two types of cholesterol, risk factors for heart disease, and the importance of diet and exercise. A total of 1,806 randomly selected subjects were interviewed in their homes in 40 geographic regions. After examining the product container with one of the three labels being tested, they were given a questionnaire to test their understanding of the label communication points. They were then asked to read the package insert and tested on their understanding of the messages it communicated. Responses were analyzed by gender, age, and education level, as well as by label format. For the education-level subgroup analysis, the high-school-nongraduate group was supplemented by 419 subjects, for a total of 2,225 subjects. Statistical analyses were performed on completed questionnaires by an independent data analysis company. The net correct response to the two primary communication objectives was 99% among the total population. Responses to questions addressing the use of concurrent medications; types of cholesterol; purpose, dosage, and preparation of the medication; and diet and exercise were also clearly understood. This study showed a high level of comprehension of the purpose and correct use of cholestyramine among both high school graduates and nongraduates. PMID- 9217588 TI - The more things change the more they stay the same. PMID- 9217589 TI - Vascular abnormalities associated with long-term cigarette smoking identified by arterial waveform analysis. AB - PURPOSE: Consistent changes in the arterial pulse contour are found with aging and disease states that impair the compliance characteristics of blood vessels that buffer pulsatile phenomena in the arterial tree. We assessed whether vascular adaptation in structure or tone of blood vessels associated with long term cigarette smoking would influence steady state or pulsatile hemodynamics at a preclinical stage. PATIENTS AND METHODS: We analyzed intraarterial brachial artery waveforms in 35 healthy long-term cigarette smokers and 32 nonsmoking control subjects matched for age and gender. The diastolic pressure decay was segmented into two components: an exponential decay that reflects the compliance characteristics of the large arteries and an oscillatory diastolic waveform generated principally by pulse-wave reflections from small arteries and arterioles. RESULTS: Resting heart rate was higher in smokers than nonsmokers, mean +/- SD (66 +/- 9 versus 60 +/- 10; P < 0.05). Systolic, diastolic, and mean arterial pressures were lower in smokers compared with nonsmokers (P < 0.01 for all). No differences in cardiac output, large artery compliance, or systemic vascular resistance estimates where apparent between groups. A decrease in the amplitude and duration of the diastolic wave, produced by peripheral pulse-wave reflections in the arterial system, was found in smokers compared with nonsmokers (0.04 +/- 0.02 versus 0.7 +/- 0.03; P < 0.001). CONCLUSIONS: Quantitative changes in the arterial waveform were found in long-term smokers compared with nonsmoking control subjects. The altered arterial wave shape marks the presence of abnormal structure or tone in the peripheral vasculature that affects pulsatile arterial function. This measure of vascular injury is detectable at a preclinical stage and may relate to the subsequent risk of morbid events in chronic smokers and aid in clinical risk stratification. PMID- 9217591 TI - Reversible hypercapnia in chronic obstructive pulmonary disease: a distinct pattern of respiratory failure with a favorable prognosis. AB - PURPOSE: Hypercapnia is regarded as a poor prognostic indicator in chronic obstructive pulmonary disease (COPD), but many patients hospitalized with hypercapnia associated with an acute exacerbation of COPD revert to normocapnia during recovery. We wished to determine if this reversible hypercapnia represents a distinct pattern of respiratory failure in COPD, or simply a stage in the progression to chronic hypercapnia. We therefore compared the long-term clinical progression and survival of COPD patients with reversible hypercapnic respiratory failure (defined as type 2.1) to those with normocapnic (PaCO2 < 50 mm Hg; type 1) and also to those patients with chronic hypercapnic (PaCO2 > 50 mm Hg) respiratory failure (defined as type 2.2). PATIENTS AND METHODS: We prospectively followed for 5 years a cohort of 85 patients who had been admitted as emergencies during a 1-year period to the respiratory unit of a University teaching hospital with an exacerbation of COPD complicated by respiratory failure (PaO2 < 60 mm Hg). The main long-term outcome measures were survival and blood gas changes. RESULTS: Sixty-eight (80%) patients survived the initial admission, and 17 (27%) survived 5 years. PaCO2 rose substantially more during exacerbations in type 2.1 patients (mean 15.8 mm Hg), compared with type 2.2 (mean 6.8 mm Hg) and type 1 patients (mean 1.5 mm Hg). We analyzed 149 subsequent admissions among the survivors over the following 5 years. Type 2.1 patients had a better 5-year survival (28%) than type 2.2 (11% survival; P < 0.05), and similar to type 1 patients (33% 5-year survival). Only 24% of reversible hypercapnic patients developed chronic hypercapnia during long-term followup. CONCLUSIONS: The data support reversible hypercapnia being a distinct manifestation of respiratory failure in COPD, with a similar prognosis to that of normocapnic respiratory failure. PMID- 9217590 TI - Cocaine effects on digital blood flow and diffusing capacity for carbon monoxide among chronic cocaine users. AB - PURPOSE: To determine the acute effects of intravenous (i.v.) cocaine on primarily digital skin blood flow and diffusion capacity for carbon monoxide (CO), and secondarily on subjective and cardiovascular measures. PATIENTS AND METHODS: A double-blind, Latin-square, placebo-controlled, dose-response study was conducted in an inpatient general clinical research center and clinical pharmacology unit of a university teaching hospital. Twelve adult males with histories of illicit drug use including i.v. cocaine received 0, 25, and 50 mg of i.v. cocaine given as 1-minute infusions, on 3 consecutive test days. Digital cutaneous blood flow was determined via laser doppler flowmetry and skin temperature. Diffusing capacity for carbon monoxide (DCO) was measured with standard techniques. Subjective responses were measured by oral report of a numerical ranking of strength of drug effect. Heart rate and blood pressure responses were measured by electronic sphygmomanometer. RESULTS: A maximal decrease in skin blood flow occurred at 2 to 3 minutes after infusion, and was not distinguished among drug conditions. Blood flow returned to baseline more rapidly after placebo than after cocaine: 7 minutes (placebo), 35 minutes (25 mg cocaine), 50 minutes (50 mg cocaine). Skin temperature decreased by 1.25 degrees C after placebo and by 2.75 and 3.25 degrees C after 25 and 50 mg of cocaine, respectively. DCO changed by -1.02 (mean) +/- 0.25 (standard deviation), 0.16 +/- 1.22, and 0.21 +/- 1.63 ml/min/mm Hg following placebo, 25, and 50 mg of cocaine, respectively. Typical subjective, chronotropic, and pressor responses to cocaine were demonstrated, and these occurred in close temporal relationship to digital blood flow and skin temperature responses. CONCLUSIONS: The digital cutaneous circulation is highly sensitive to vasoconstrictor effects of cocaine. Pulmonary blood volume tends to be preserved after i.v. cocaine. Subjective effects and cardiovascular responses occur in concert with peripheral blood flow changes. The peripheral vasoconstrictor effects have implications for cocaine users with concurrent vasospastic or vasculopathic disorders. PMID- 9217592 TI - Coronary vasomotor reactivity among normotensive African and white American subjects with chest pain. AB - BACKGROUND AND OBJECTIVES: Excess cardiovascular morbidity and mortality among African (black) Americans is the subject of intensive investigation but the etiology remains speculative. One hypothesis proposes that inherent, or intrinsic, differences in coronary vascular reactivity and endothelial function predispose African Americans to enhanced vasoconstriction and/or depressed vasodilation, resulting in excess ischemia. The objective of this study was to establish whether coronary vasoreactivity differs among normotensive, nondiabetic African and white Americans with normal arteries referred for coronary arteriography because of chest pain. PATIENTS AND METHODS: Eleven African American (8 female, 3 male) and 28 white American (9 female, 19 male) normotensive, euglycemic patients with normal coronary arteries were prospectively recruited for invasive testing of coronary artery and microvascular relaxation using the endothelium-dependent and -independent agents, acetylcholine and adenosine; a Doppler tipped intracoronary guidewire; and quantitative coronary angiography. RESULTS: The study cohort consisted of 17 women (44%) and 22 men (56%) with a mean age of 46 +/- 10 yrs. Of 8 African American women, 6 were premenopausal and 2 were postmenopausal on estrogen replacement therapy. Of 9 white American women, 2 were premenopausal, 1 was 46-year old with a previous history of hysterectomy without ovariectomy, 2 were postmenopausal on estrogen replacement therapy, 2 were perimenopausal and 44- and 54-year old, and 2 were postmenopausal without estrogen replacement therapy. In response to maximal infusion of acetylcholine, epicardial coronary arteries and resistance vessels dilated similarly in black and white subjects. Dose-response curves revealed no significant racial differences during submaximal graded infusion of acetylcholine. In response to peak effect of adenosine, there were no racial differences in dilation of the microcirculation. CONCLUSIONS: In the absence of hypertension, diabetes mellitus, and angiographic evidence of coronary artery disease, African American women demonstrate no evidence of intrinsic predisposition to enhanced coronary conduit vasoconstriction or depressed microcirculatory dilation in response to the endothelium-dependent and independent vasodilator agonists-acetylcholine and adenosine-when compared with responses of similar white men and women. Because of low enrollment of black males, definitive conclusions cannot be drawn regarding this group. PMID- 9217593 TI - Cardiac valvular vegetations in cancer patients: a prospective echocardiographic study of 200 patients. AB - PURPOSE: Nonbacterial thrombotic endocarditis can complicate various malignancies and may cause morbidity and mortality mainly as a result of systemic embolism. The antemortem diagnosis of nonbacterial thrombotic endocarditis is rare. The purpose of our study was to assess the frequency, echocardiographic characteristics, and clinical correlation of nonbacterial thrombotic endocarditis in cancer patients. PATIENTS AND METHODS: A prospective echocardiographic screening of 200 nonselected ambulatory patients with solid tumors was performed. Patients were evaluated for evidence of thromboembolic events and for plasma D dimer levels. A cohort of 100 consecutive patients without overt heart disease referred to echocardiography for the detection of an occult arterial embolic source served as a control group. It consisted of 52 males and 48 females, median age 60 years. RESULTS: The study group included 87 women and 113 men, median age 64 years (range 21 to 91). The frequent malignancies were lymphoma (26%), carcinoma of the gastrointestinal tract (20%), and carcinoma of the lung (16%). Cardiac valvular vegetations were found in 38 patients (19%) compared with only in 2 patients in the control group (2%, P < 0.001). Vegetations were found on the mitral or on the aortic valve in 19 and 18 patients, respectively. Isolated tricuspid valve vegetation was found in 1 patient. Valvular lesions were mostly common in patients with carcinoma of the pancreas (3 of 6, 50%), carcinoma of the lung (9 of 32, 28%), and lymphoma (10 of 52, 19%). Thromboembolism was diagnosed in 22 (11%) patients (12 deep vein thrombosis, 4 emboli to extremities, 2 cerebrovascular accidents, and 4 "silent" segmental left ventricular wall motion abnormalities on echocardiography). Thromboembolism was noticed in 9 of 38 patients (24%) with vegetations compared with 13 of 162 patients without vegetations (8%; P = 0.013). Plasma D-dimer level was examined in a subgroup of 170 patients. D-dimer level was increased in 19 of 21 patients (90%) with thromboembolism compared with 76 of 149 patients without thromboembolism (51%; P = 0.001). CONCLUSIONS: This study demonstrated a high prevalence of cardiac valvular lesions in patients with solid tumors. Vegetations were associated with thromboembolism. Plasma D-dimer level was significantly increased in patients with thromboembolism. PMID- 9217594 TI - Divergent trends in obesity and fat intake patterns: the American paradox. AB - PURPOSE: To compare recent changes in diet and physical activity with trends in body weight and obesity prevalence, using large survey studies representative of the US population. MATERIALS AND METHODS: Secular-trends survey studies were made from databases of NHANES II and III, USDA Nationwide Food Consumption Survey, Behavioral Risk Factor Survey System, and Calorie Control Council Report providing data on obesity prevalence, body mass index, calorie and fat intake, exercise-related physical activity, and consumption of low-calorie food extracted from surveys for the adult US population and specific subgroups. RESULTS: In the adult US population the prevalence of overweight rose from 25.4% from 1976 to 1980 to 33.3% from 1988 to 1991, a 31% increase. During the same period, average fat intake, adjusted for total calories, dropped from 41.0% to 36.6%, an 11% decrease. Average total daily calorie intake also tended to decrease, from 1,854 kcal to 1,785 kcal (-4%). Men and women had similar trends. Concurrently, there was a dramatic rise in the percentage of the US population consuming low-calorie products, from 19% of the population in 1978 to 76% in 1991. From 1986 to 1991 the prevalence of sedentary lifestyle represented almost 60% of the US population, with no change over time. CONCLUSIONS: Reduced fat and calorie intake and frequent use of low-calorie food products have been associated with a paradoxical increase in the prevalence of obesity. These diverging trends suggest that there has been a dramatic decrease in total physical activity related energy expenditure. Efforts to increase the average American's total exercise- and nonexercise-related physical activities may be essential for the prevention of obesity. PMID- 9217595 TI - Impact of obesity on allogeneic stem cell transplant patients: a matched case controlled study. AB - PURPOSE: To determine the impact of obesity on survival after high-dose therapy followed by allogeneic stem cell transplant in adults and children with various malignancies as well as metabolic disorders. PATIENTS AND METHODS: A matched case controlled evaluation of 322 allogeneic patients from a single institution with a median follow-up of 296 and 120 days among nonobese and obese patients, respectively, was conducted between April 1983 and June 1995 at the University of Kentucky. The overall survival distributions among subsets defined as either obese or nonobese were measured. RESULTS: The overall survival among the nonobese and obese was 35% and 20%, respectively (P = 0.0045). When patients were separated by age, the adult patients maintained this difference, while the children did not. When patients were stratified according to donor status, both the histocompatible and the nonhistocompatible adults had an inferior outcome among obese patients. The difference, however, was significant only among the histocompatible group (P = 0.0007). Causes of deaths were insignificantly distributed among both relapse as well as nonrelapse mechanisms. CONCLUSION: Adult obese patients undergoing high-dose chemotherapy with stem cell rescue have a more adverse outcome. Both relapse and nonrelapse causes are responsible for the different outcome between obese and nonobese groups. PMID- 9217596 TI - Pain during hospitalization is associated with continued pain six months later in survivors of serious illness. The SUPPORT Investigators. Study to Understand Prognoses and Preferences for Outcomes and Risks of Treatments. AB - PURPOSE: To determine the level of pain reported by survivors of serious illness 2 and 6 months after study enrollment and to identify variables associated with later pain. PATIENTS AND METHODS: Observational cohort study of patients with interviews during hospitalization (5,652) and 2 (3,782) and 6 (2,984) months later admitted between June 1989 and January 1994 with 1 or more of 9 high mortality diagnoses admitted to 5 tertiary care academic centers in the United States. Patients' level of pain during the hospitalization and 2 and 6 months later was determined from interviews with patients and surrogates (most often family members). Separate ordinal logistic regressions were constructed with level of pain at months 2 or 6 as the dependent variable and 22 demographic, psychological, chronic, and acute illness measures at the time of hospitalization as independent variables. RESULTS: Of patients reporting level 4 (moderately severe pain occurring most of the time or extremely severe pain occurring half of the time) or 5 (moderately severe pain occurring most or all of the time or extremely severe pain occurring at least half of the time) pain to 5 during hospital interviews, 39.5% and 39.7% reported level 4 or 5 pain 2 and 6 months later, respectively. Level of hospital pain was the variable most strongly associated with later pain. Compared with patients with level 1 hospital pain, those with level 2 (not at all severe pain or moderate, occasional) had a 2.91 (95% confidence interval [CI] 2.50, 3.37) and 1.75 (CI 1.48, 2.07) times greater adjusted odds of increased levels of pain 2 and 6 months later, respectively. Compared with patients with level 1 hospital pain, those with level 5 pain had a 9.20 (CI 7.27, 11.65) and 4.40 (CI 3.39, 5.71) times greater adjusted odds of increased levels of pain 2 and 6 months later, respectively. Age, number of dependencies in activities of daily living, depression, and type of comorbid illnesses were also independently associated with level of pain both 2 and 6 months later. CONCLUSION: Survivors of the serious and common illnesses that we studied have a high level of pain during hospitalization and up to 6 months after hospitalization. Level of hospital pain was most strongly associated with later pain. Better pain control both during hospitalization and after discharge should be given a high priority. Pain during hospitalization should trigger future inquiries about pain and its treatment. PMID- 9217597 TI - Body mass index as a correlate of postoperative complications and resource utilization. AB - PURPOSE: To describe the relationship of body mass index (BMI) with postoperative complications and resource utilization. PATIENTS AND METHODS: Two thousand nine hundred and sixty-four patients 50 years or older undergoing elective noncardiac surgery with an expected length of stay > or = 2 days were enrolled in a prospective cohort study to measure major cardiac complications, noncardiac complications, length of stay, and costs. The setting was an urban teaching hospital. A preoperative history, physical, electrocardiogram (ECG), and chart review were performed by study personnel. Postoperative complications were detected by ECGs, creatine kinase and creatine kinase MB levels, and daily chart review. Total costs were obtained from the hospital's computerized database. RESULTS: Complication rates were not different among BMI groups (underweight < 20, normal 20 to 29, overweight 30 to 34, most overweight > 34), but patients with BMI 30 to 34 and > 34 who underwent abdominal or gynecologic procedures had significantly higher wound infection rates (11% each) than normal weight patients (4.7%) or the underweight (0%). After adjusting for age, race, gender, smoking history, comorbid diseases, procedure type, and insurance status, there were nonsignificant trends toward increased resource utilization by the most overweight patients (BMI > 34). These patients stayed 0.8 days longer (P = 0.13) and had total costs that were $843 higher (P = 0.17) than patients of normal weight (BMI 20 to 29). The underweight patients stayed 0.9 days longer (P = 0.23) and had total costs that were $3,150 higher (P = 0.04) than patients of normal weight. Quadratic models to test for a U-shaped relationship found no correlation between BMI and length of stay, but did find that BMI was significantly correlated with total costs (P = 0.04). This relationship persisted when patients who had complications were excluded from the analysis. CONCLUSIONS: Overall, BMI was not significantly correlated with postoperative complications or length of stay. However, overweight patients who underwent abdominal or gynecologic procedures had higher wound infection rates, and patients with the highest and lowest BMIs had significantly higher adjusted total costs. PMID- 9217598 TI - Vancomycin-resistant enterococci. AB - Enterococci have been recognized as an important cause of nosocomial infections for almost 20 years and as a cause of endocarditis for almost a century. While long known for their capacity of displaying multiple antibiotic resistant traits, the extent to which this could occur was not fully appreciated until the emergence of enterococci with acquired resistance to vancomycin; this resistance has been particularly problematic because it often occurs in the uncommon subset of enterococci that are also highly resistant to ampicillin-a combination with devastating therapeutic consequences. The observation that vancomycin resistance can be transferred to and expressed in other gram-positive organisms, for which vancomycin is often considered the primary therapeutic alternative, is a chilling reminder of just how close we may be to a wide array of potentially untreatable "killer" microbes. PMID- 9217599 TI - Euthanasia: an unbiased decision? PMID- 9217600 TI - Tuberculous brain abscess in a patient with HIV infection: case report and review. PMID- 9217601 TI - Vasculitis due to cholesterol embolism. PMID- 9217602 TI - Failure of cefotaxime treatment in an adult with Streptococcus pneumoniae meningitis. PMID- 9217603 TI - Scabies mimicking systemic lupus erythematosus. PMID- 9217604 TI - Spontaneous rupture of popliteal artery cyst with relief of claudication. PMID- 9217605 TI - My hope for medicine. PMID- 9217606 TI - Fever and renal failure in a 31-year-old male with AIDS. PMID- 9217607 TI - Antiphospholipid antibodies and essential thrombocythemia. PMID- 9217608 TI - Antiphospholipid antibodies and thrombosis. PMID- 9217609 TI - IgM monoclonal gammopathy, lymphoid proliferations and lupus anticoagulant. PMID- 9217610 TI - Low-dose dopamine in acute oliguric renal failure. PMID- 9217611 TI - Clinical trials in departments of internal medicine: ingredients for success. PMID- 9217612 TI - Studies show that it is now time to vigorously promote screening for colorectal cancer. PMID- 9217613 TI - Risk factors and predictive models of giant cell arteritis in polymyalgia rheumatica. AB - OBJECTIVE: To identify in polymyalgia rheumatica the best set of predictors for a positive temporal artery biopsy and to define predictive models with either a high or low probability of giant cell arteritis (GCA). PATIENTS AND METHODS: Retrospective study of 227 patients, 137 with polymyalgia rheumatica unassociated with arteritis (group A) and 90 with polymyalgia associated with biopsy-proven giant cell arteritis (group B or training set). Data on demographic features, clinical and laboratory abnormalities were collected. Risk factors for arteritis were estimated by nonlinear logistic regressions. Simple predictive models were constructed with those predictors more related to arteritis by multivariable analysis. These models were then tested in group B and in 89 cases of arteritis without polymyalgia rheumatica (group C or test set). RESULTS: The best predictors of arteritis were a new headache odds ratio (OR) 13.6 (95% confidence interval [CI] 4.7 to 39.3); age at onset < 70 years OR 0.11 (CI 0.04 to 0.35); abnormal temporal arteries OR 4.2 (CI 1.3 to 13.7); raised liver enzymes OR 2.9 (CI 1.1 to 7.8), and jaw claudication OR 4.8 (CI 1.0 to 22.7). Amaurosis was only observed in patients with arteritis. Three subsets had a very high risk of arteritis: (1) Patients with recent headache, abnormal arteries, and > or = 70 years at disease onset: sensitivity 44%, positive predictive value (PPV) 93%, likelihood ratio (LR) 20.3; (2) patients with a new headache, jaw claudication, and abnormal arteries: sensitivity 34.4%, PPV 96.9%, LR 47.2; and (3) those, that in addition to the last 3 features, were > or = 70 years of age at disease onset: sensitivity 26.7%, PPV 100%. We could also identify a subset with a very low risk of arteritis constituted by patients < 70 years, without headache, and with clinically normal temporal arteries: sensitivity 1.1%, PPV 1.7%, LR 0.03. In group C or the test set, these four predictive models correctly identified 57.3%, 29.2%, 23.6, and 3.4% of patients, respectively. CONCLUSIONS: In polymyalgia rheumatica it is feasible to identify subsets with a very high likelihood of GCA. Although in some of these subsets the diagnosis of arteritis is almost certain, we suggest that even then it should be confirmed by temporal artery biopsy. By contrast, in those patients with polymyalgia < 70 years and without cranial features of giant cell arteritis, the risk of vasculitis is so low that the biopsy could be initially avoided and the patient treated with low-dose corticosteroids. PMID- 9217614 TI - The clinical impact of echocardiography on antibiotic prophylaxis use in patients with suspected mitral valve prolapse. AB - PURPOSE: To determine the impact of echocardiography on the use of antibiotic prophylaxis in patients with suspected mitral valve prolapse (MVP). PATIENTS AND METHODS: We evaluated 147 consecutive patients who were referred for "rule out mitral valve prolapse" to a university hospital echocardiography laboratory. Chart review and phone contact were used to determine the demographic characteristics of the patients; past diagnosis of MVP, symptoms, and exam at referral; practice specialty of referring MD; echocardiographic findings; and change in prophylaxis usage as a result of the echocardiogram (ECHO). Prophylaxis was considered to be indicated if the echocardiogram demonstrated MVP with at least mild regurgitation or abnormal thickening of at least one mitral leaflet. RESULTS: Based on the ECHO a change in antibiotic prophylaxis was indicated in 20 of 147 (14%) patients including initiation of prophylaxis in 6, and discontinuation of prophylaxis in 14. However, only 4 of 20 patients (20%) actually changed their prophylaxis habits leading to an actual yield of 4 management changes per 131 ECHOs ordered (3%). This corresponded to 1 change in management per $36,250 in hospital and physician costs. Younger age, female gender, and presence of symptoms were associated with a benign ECHO. Indications for a change in management were not significantly different between physician specialities: 18% for generalists (internal medicine and family practice), 12% for cardiologists, and 7% for other specialists, P = 0.3. CONCLUSIONS: In patients referred for evaluation of MVP, echocardiography infrequently resulted in changes in antibiotic prophylaxis management and was associated with significant expense. PMID- 9217615 TI - Prognostic significance of coronary calcific deposits in asymptomatic high-risk subjects. AB - PURPOSE: To determine the predictive value of coronary calcifications for coronary heart disease events in high-risk, asymptomatic adults: PATIENTS AND METHODS: A prospective cohort study of 1,461 high-risk, asymptomatic subjects were followed for 55 months with a 98% success rate. Coronary risk factor assessment and cardiac fluoroscopy with digital subtraction enhancement were performed to determine the number of calcified coronary arteries. RESULTS: Fifty eight percent of this cohort (852 subjects) had fluoroscopically detectable coronary calcification: 437 (30%) had calcium in one, 253 (17%) in two, and 162 (11%) in all three coronary vessels. There were 90 (6%) deaths, 35 (39%) attributable to coronary heart disease, and 43 (3%) nonfatal myocardial infarctions. Subjects with calcification in more than one major coronary artery were 2.2 times more likely to suffer coronary death or nonfatal infarction (P = 0.001) than were subjects with one or no calcified arteries. Multivariable logistic regression analysis showed that only the number of calcified arteries, age, total cholesterol, history of diabetes, and left ventricular hypertrophy by electrocardiogram were associated independently with the incidence of coronary death or infarction in these subjects. CONCLUSIONS: Coronary calcification predicts coronary heart disease death or infarction in high-risk asymptomatic adults as well as do standard risk factors. PMID- 9217616 TI - Development and validation of a clinical score to estimate the probability of coronary artery disease in men and women presenting with suspected coronary disease. AB - PURPOSE: Guidelines for the management of patients with suspected coronary disease have emphasized stratification into groups with low, intermediate, and high probability of significant coronary disease. Previously derived clinical prediction rules have been difficult to apply in clinical settings. The purpose of this study was to develop and validate a clinical score that facilitates this stratification process. PATIENTS AND METHODS: We performed a retrospective analysis of prospectively acquired data from 915 patients with suspected coronary disease and normal resting electrocardiograms who presented for exercise testing at a university hospital. All patients subsequently underwent coronary angiography. Analysis included logistic regression with significant coronary disease (> or = 1 vessel with a > or = 50% lesion) presence as the dependent variable and clinical variables as independent variables. From this analysis, a coronary disease score was developed to estimate prevalence of coronary disease from clinical variables. Validation of this score was performed in a separate prospectively acquired cohort of 348 patients. RESULTS: For the entire validation group, the prevalence of significant coronary disease was 16% (10/63) in the low probability group, 44% (86/195) in the intermediate probability group, and 69% (62/90) in the high probability group. Both men and women were stratified equally well into the 3 probability groups. CONCLUSION: The clinical score is an easily memorized and accurate method for categorizing patients with suspected but not proven coronary disease and normal resting electrocardiograms into clinically meaningful probability groups upon which decisions concerning appropriate diagnostic test selection could potentially be based. PMID- 9217617 TI - Does the chronic fatigue syndrome involve the autonomic nervous system? AB - PURPOSE: To investigate the role of the autonomic nervous system in the symptoms of patients with chronic fatigue syndrome (CFS) and delineate the pathogenesis of the orthostatic Intolerance and predisposition to neurally mediated syncope reported in this patient group. PATIENTS AND METHODS: Twenty-three CFS patients and controls performed a battery of autonomic function tests. The CFS patients completed questionnaires pertaining to autonomic and CFS symptoms, their level of physical activity, and premorbid and coexisting psychiatric disorders. The relationship between autonomic test results, cardiovascular deconditioning, and psychiatric disorders was examined with multivariate statistics and the evidence that autonomic changes seen in CFS might be secondary to a postviral, idiopathic autonomic neuropathy was explored. RESULTS: The CFS subjects had a significant increase in baseline (P < 0.01) and maximum heart rate (HR) on standing and tilting (both P < 0.0001). Tests of parasympathetic nervous system function (the expiratory inspiratory ratio, P < 0.005; maximum minus minimum HR difference, P < 0.05), were significantly less in the CFS group as were measures of sympathetic nervous system function (systolic blood pressure decrease with tilting, P < 0.01; diastolic blood pressure decrease with tilting, P < 0.05; and the systolic blood pressure decrease during phase II of a Valsalva maneuver, P < 0.05). Twenty-five percent of CFS subjects had a positive tilt table test. The physical activity index was a significant predictor of autonomic test results (resting, sitting, standing, and tilted HR, P < 0.05 to P < 0.009); and the blood pressure decrease in phase II of the Valvalsa maneuver, P < 0.05) whereas premorbid and coexistent psychiatric conditions were not. The onset of autonomic symptoms occurred within 4 weeks of a viral infection in 46% of patients-a temporal pattern that is consistent with a postviral, idiopathic autonomic neuropathy. CONCLUSION: Patients with CFS show alterations in measures of sympathetic and parasympathetic nervous system function. These results, which provide the physiological basis for the orthostatic intolerance and other symptoms of autonomic function in this patient group, may be explained by cardiovascular deconditioning, a postviral idiopathic autonomic neuropathy, or both. PMID- 9217618 TI - Comparative assessment of peripheral sympathetic function by postural vasoconstriction arteriolar reflex and sympathetic skin response in NIDDM patients. AB - PURPOSE: The aim of the study was to compare peripheral sympathetic adrenergic and cholinergic nerve function in NIDDM (non-insulin-dependent diabetes mellitus) patients with various degrees of diabetic neuropathy and neuropathic foot ulceration. The parameters used were postural vasoconstriction arteriolar reflex (VAR) and sympathetic skin response (SSR), respectively. PATIENTS AND METHODS: Forty-seven NIDDM patients were studied. No patients had clinically significant peripheral vascular disease. They were divided according to peripheral somatic neuropathy, assessed by clinical score and vibration perception threshold (VPT). Twenty-two patients showed no significant evidence of peripheral neuropathy and normal VPT (DN-); 15 had signs and symptoms of neuropathy and VPT alteration (DN+); 10 had diabetic neuropathy and foot ulceration (DNU). Twenty-two normal subjects were also examined as a control group. Resting arteriovenous shunt skin blood flow, measured using laser-Doppler flowmetry, and the VAR of the big toe on lowering the foot were studied. Sympathetic skin response was assessed by an EMG apparatus. Autonomic function was also investigated by using standard cardiovascular reflex tests. RESULTS: Resting blood flow values were similar in the three NIDDM groups and in the control group. VAR to foot lowering was significantly impaired in all NIDDM groups by comparison with controls (72.8 +/- 2.1%, mean +/- SEM), this impairment being progressively more pronounced in DN- (58.8 +/- 2.3%, P < 0.001), DN+ (33.3 +/- 3.0%, P < 0.001 versus DN-) and DNU (8.6 +/- 2.7%, P < 0.001 versus DN+). Sympathetic skin response was assessed in 28 patients and was significantly impaired in DN-compared with the control group (2.53 +/- 0.04 versus 2.71 +/- 0.04 log mcV, P < 0.01). This impairment was severe in the DNU compared with the DN+ group (1.36 +/- 0.05 versus 2.26 +/- 0.04 log mcV, P < 0.005). A positive correlation was found between VAR values and SSR (P < 0.001), and these measurements were also closely correlated with several parameters of central autonomic and somatic neuropathy. CONCLUSION: These results indicate that peripheral sympathetic adrenergic and cholinergic fibers simultaneously undergo early alterations in diabetic patients, even when there is no clinical neuropathy. Our data also show almost complete abolition of peripheral sympathetic activity in NIDDM patients with foot ulceration. PMID- 9217619 TI - A randomized trial of office-based screening for common problems in older persons. AB - PURPOSE: To test the effectiveness of a 10-minute office-staff administered screen to evaluate malnutrition/weight loss, visual impairment, hearing loss, cognitive impairment, urinary incontinence, depression, physical limitations, and reduced leg mobility among older persons seen in office practice. This screen was coupled with clinical summaries to assist the physician in further evaluating and managing the screen-included problems. PATIENTS AND METHODS: Twenty-six community based office practices of internists and family physicians in Los Angeles were randomized to intervention or control groups. Two hundred and sixty-one patients aged > or = 70 years and seeing these physicians for a new visit or a physical examination participated in the study. At the enrollment visit intervention group patients were administered the screening measure and their physicians were given the pertinent clinical summaries. Outcome measures were detection of, and intervention for conditions screened, and health status 6 months after the intervention. RESULTS: Hearing loss was both more commonly detected (40% intervention versus 28% control) and further evaluated (29% versus 16%) by physicians in the intervention group (P < 0.05). No other differences in the frequency of problem detection or intervention were noted between groups. Six months after the intervention no differences were noted in health status between groups. CONCLUSIONS: A brief measure to screen for common conditions in older persons was associated with more frequent detection and follow-up assessment of hearing loss. Although the measure was well accepted by physicians and their staffs, it did not appear to affect detection and intervention in regard to the other screen-included conditions, or health status at 6 months. PMID- 9217620 TI - Neurologic manifestations in Staphylococcus aureus endocarditis: a review of 260 bacteremic cases in nondrug addicts. AB - PURPOSE: To investigate the neurologic manifestations of infective endocarditis caused by Staphylococcus aureus in a population of nondrug addicts with special emphasis on the clinical presentation, epidemiology, and mortality. PATIENTS AND METHODS: During the period from 1982 to 1991 a total of 8,514 cases of bacteremia with S aureus were reported to the Staphylococcus Laboratory, Copenhagen, Denmark. The medical records of cases of suspected infective endocarditis were retrospectively reviewed and classified according to the new diagnostic criteria for endocarditis proposed by Durack. RESULTS: A total of 260 cases from 63 hospitals fulfilled the diagnostic criteria. Overall, 91 patients (35%) experienced neurologic manifestations. Sixty-one presented with neurologic symptoms, whereas 30 patients developed neurologic complications at various intervals (median: 10 days) after the debut of the disease. The most frequent neurologic manifestation was unilateral hemiparesis, which occurred in 41 patients (45%). Forty-two percent of the females had neurologic manifestations compared to only 30% of the males (P = 0.06). Cases with native mitral valve infection had a significantly higher frequency of neurologic manifestations compared with all other valvular involvement (44% versus 29%, P = 0.02) but the frequency of neurologic complications was only nonsignificantly higher in those patients with native mitral valve infection than in those patients with native aortic valve infection (44% versus 31%, P = 0.10). Only two of the patients with tricuspid valve infection and none of those with congenital heart disorder experienced neurologic manifestations. A neurologic manifestation occurred in 22 (35%) of the 63 episodes in which vegetations were detected on the echocardiograms, compared with 17 (26%) of the 65 episodes without vegetations (P = 0.38). The mortality was 74% in patients with major neurologic manifestations and 56% in patients without neurologic manifestations (P = 0.008). In patients with neurologic complications the mortality was significantly higher among those treated with antibiotics alone as compared with those treated surgically (65 of 81, 80% versus 2 of 10, 20%; P = 0.0003). CONCLUSIONS: In a population of nondrug addicts with infective endocarditis caused by S aureus the following main conclusions can be drawn: neurologic manifestations occur with a higher frequency in patients with native mitral valve infection. The presence of vegetations on echocardiograms is not a risk factor for developing neurologic complications but this conclusion is based on the results of transthoracic echocardiograms performed in only one half of the patients. The majority of the neurologic manifestations occur on presentation or shortly thereafter and the risk of recurrent embolism is low. Mortality is increased in patients with neurologic manifestations. A neurologic event per se may constitute an indication for surgical treatment. PMID- 9217621 TI - The efficacy of lovastatin in lowering cholesterol in African Americans with primary hypercholesterolemia. AB - PURPOSE: To evaluate the efficacy of lovastatin in African Americans (AA) diagnosed with primary hypercholesterolemia. PATIENTS AND METHODS: Forty-seven AA patients from the King/Drew Medical Center in Los Angeles were recruited from the Hypertension, Family Practice, and General Medicine Clinics for a double-blinded, placebo-controlled trial. Forty-one patients completed the 10 week study. Eligibility for entrance into the study was determined by patient lipid profiles meeting the criteria for pharmacological intervention outlined by the National Cholesterol Education Program II guidelines. Patients were randomized into 2 groups: lovastatin 20 mg per day, or placebo. A registered dietitian counseled both groups on two visits during the study to ensure compliance with a low fat, low cholesterol diet. Lipid levels were compared at the first and last visit of the study. RESULTS: The lovastatin-treated group demonstrated significant reductions in mean total cholesterol (TC) (14.7%, 95% confidence interval [CI] 6.6 to -22.8, P < 0.01) and low-density lipoprotein (LDL) cholesterol (20.0%, 95% CI-7.9 to -32.1, P < 0.01) from baseline. Plasma triglyceride (TG) levels decreased by 10.5% (95% CI-2.4 to -18.6) and total cholesterol/high density lipoprotein (HDL) ratio fell below five in the lovastatin group, but neither reduction reached statistical significance. Placebo administration was not associated with any significant changes in TC, LDL, or TG. There were no significant differences between baseline and post-treatment hepatic transaminase levels in either group. CONCLUSIONS: The HMG-CoA (3-hydroxyl-3 methylglutary coenzyme A) reductase inhibitor lovastatin in a dose of 20 mg per day was effective in decreasing TC, LDL, and TG levels in an AA population. Considering that the AA population is at substantially increased risk for hypertension and cardiovascular morbidity, more aggressive and wider use of HMG-CoA reductase inhibitors should be employed in reducing elevated plasma cholesterol levels. PMID- 9217622 TI - Management of early diabetic nephropathy. PMID- 9217623 TI - Asking patients what they like: overlooked attributes of patient satisfaction with primary care. AB - PURPOSE: Ask patients to describe important attributes of their primary health care; and use the responses to develop a taxonomy for classifying patient satisfaction. DESIGN: Open-ended questions were administered to patients immediately after a clinic visit. SETTING: Primary care clinics at an academically affiliated Veterans Affairs Medical Center in New England. PATIENTS: Two hundred two of 204 randomly selected English-speaking patients who agreed (and were able) to participate. INTERVENTIONS: Clinimetric methods were used to obtain responses to three open-ended questions about what patients liked, disliked, and would like to see changed about their care. These "raw" descriptions were then combined into pertinent groups and arranged as a taxonomy of patient satisfaction. RESULTS: The taxonomy was divided into five main axes referring to physician staff, nonphysician staff, attributes of the clinic, related services, and the institution. The axes contained a total of 34 items related to patient satisfaction. The items have demonstrable face validity, and are likely to be "transparently" sensible to clinicians and policy makers, but many of the items-such as problems with parking-were not included in either of two existing psychometric instruments used to measure patient satisfaction in the same clinics. CONCLUSIONS: The clinimetric strategy leads to a simple, clinically relevant, and easily understood assessment of patient satisfaction with health care services. The assessment can be done with three simple questions; and the responses can be catalogued, when desired, in a suitable taxonomy. PMID- 9217625 TI - Babesiosis in a patient with sickle cell anemia. PMID- 9217624 TI - A review of hereditary breast cancer: from screening to risk factor modification. AB - The identification of genetic mutations thought to be directly responsible for the development of breast cancer represents a major advance in our understanding of this disease. Mutations in BRCA1 and BRCA2 are thought to be responsible for the majority of inherited breast cancer. Although these mutations account for approximately 5% of breast cancer cases, the identification of these genes will have a profound impact on the way patients and their physicians view breast cancer risk. Genetic testing for BRCA1 and BRCA2 mutations is already available. Interpreting results of genetic tests for these mutations is problematic and the clinical management of women carrying these gene mutations is far from straightforward. The purpose of this paper is to review recent developments in the genetic aspects of breast cancer, including genetic testing, to critically review risk factor modification, and to discuss screening and potential prophylactic measures. PMID- 9217626 TI - Estrogen-induced pancreatitis after discontinuation of concomitant medroxyprogesterone therapy. PMID- 9217627 TI - A case report of carotid artery dissection presenting as cluster headache. PMID- 9217628 TI - Etiology of chronic fatigue syndrome. PMID- 9217629 TI - Strategic business planning for internal medicine. PMID- 9217630 TI - Plasma thrombomodulin in atherosclerosis and its risk factors. PMID- 9217631 TI - End-stage renal disease in systemic lupus erythematosus. PMID- 9217632 TI - Epidemiology of acquired immunodeficiency syndrome: advancing to an endemic era. AB - Acquired Immunodeficiency syndrome (AIDS) is now endemic in the United States. Human immunodeficiency virus (HIV) is the leading cause of death in U.S. adults aged 25-44 years. While the rate of infection among men who have sex with men has stabilized, that among women, minorities, and young people is increasing. Improved methods for identifying these subpopulations at risk are needed. More than half the people infected with HIV do not know that they are. Recent studies demonstrate rapid viral replication during clinically asymptomatic disease and support the need for early initiation of combination antiretroviral therapy to slow disease progression and to improve patient longevity. However, significant numbers of individuals, particularly those at highest risk, continue to be diagnosed late in their illness. A recently approved oral test for HIV-1 samples mucosal transudate and provides a less invasive, but equally reliable, alternative to blood testing that may increase the acceptability and accessibility of HIV testing. Nonetheless, it remains to be determined whether this oral testing approach will have a significant public health impact among risk groups with low access to screening and care. PMID- 9217633 TI - Oral diagnostic testing for detecting human immunodeficiency virus-1 antibodies: a technology whose time has come. AB - An oral fluid-based test for antibodies to human immunodeficiency virus (HIV), equivalent to serum in its accuracy but safer and easier to use, is now available in the United States. The development of the oral test involved overcoming technical obstacles to the use of oral fluid as a testing medium, including low immunoglobulin G (IgG) titers, suboptimal assay performance, protease degradation of IgG, high viscosity, and lack of a standardized method of specimen collection, all of which contribute to suboptimal assay performance. The currently available oral HIV test utilizes a collection device to isolate a mucosal transudate component of oral fluid rich in IgG. A vial containing a preservative solution facilitates the transport of stable, low-viscosity specimens to the laboratory for testing with an ELISA and confirmatory Western blot assay, specifically designed for use with oral fluid. Non-HIV medical conditions and oral pathologies do not appear to affect oral test results. Hopefully, the availability of a more patient-friendly, portable diagnostic test for antibodies to HIV will facilitate identification of greater numbers of infected individuals with the ultimate goals of early identification, early treatment, and prevention of disease transmission. PMID- 9217634 TI - Accuracy of oral specimen testing for human immunodeficiency virus. AB - Antibodies to human immunodeficiency virus (HIV) can be detected in oral fluid with great accuracy, due to technical advances in both the collection of oral samples and assays. Reported sensitivities of 97.2-100% and specificities of 97.7 100% compare well with those of serum-based assays and qualify oral fluid for the screening and diagnosis of HIV infection in both high- and low-risk populations. In addition, oral fluid offers several advantages over serum as a testing medium for HIV, including greater safety, ease of collection, and patient acceptability. PMID- 9217635 TI - Future applications of oral fluid specimen technology. AB - Research has demonstrated that oral mucosal transudate (OMT), a serum-derived fluid that enters saliva from the gingival crevice and across oral mucosal surfaces, can be preferentially concentrated by a novel collecting system to yield detectable levels of immunoglobulins (i.e., IgG and IgM antibodies) against various bacterial and viral diseases. Assays based on OMT can aid in the diagnosis of disease and in the management of therapeutic drugs. A reliable and accurate OMT-based test to detect human immunodeficiency virus (HIV) antibodies is commercially available. Additional tests based on similar technologies may aid in the diagnosis of viral hepatitis, measles, mumps, and rubella as well as in monitoring levels of therapeutic drugs such as theophylline. The future use of OMT-based testing will likely increase because of the inherent advantages of this technology: convenience; avoidance of inadvertent transmission of blood-borne pathogens; ease of use in pediatric and geriatric populations; as well as the potential for blood-free home and workplace collection of patient samples. PMID- 9217636 TI - Virucidal activity of an oral fluid human immunodeficiency virus-1 antibody preservative. AB - Diagnosis of human immunodeficiency virus-1 (HIV-1) infection requires the collection of either serum or oral fluid that is subsequently tested for the presence of antibodies to HIV-1. The effective use of oral fluid for the detection of HIV antibodies is contingent on stabilization of immunoglobulins in the sample through the use of preservatives. Oral fluid preservatives also contain agents that can disrupt and inactivate viruses. This study demonstrates the virucidal activity of a commercially available oral fluid preservative against HIV-1 using a sensitive 28-day cell culture assay designed to detect infectious virus. The results demonstrate that a 5-log reduction in viral titer is obtained when equal volumes of HIV-1 viral stocks and the preservative are mixed. The data provide strong evidence that preserved oral fluid samples from infected individuals are noninfectious for HIV-1. PMID- 9217637 TI - Evidence-based medicine: a powerful educational tool for clerkship education. PMID- 9217638 TI - Population-based approaches to health care: a way to improve health for select groups. PMID- 9217639 TI - Screening in special populations: a "case study" of recent Vietnamese immigrants. AB - PURPOSE: To determine how the medical and social profile of a particular special population, Vietnamese immigrants, should be used to tailor screening protocols that differ from those designed for the general population. PATIENTS AND METHODS: A consecutive series of Vietnamese immigrants living in the United States for less than 6 months were evaluated by interviewer-administered standardized questionnaire and medical record review. A total of 99 new Vietnamese immigrants (47 women and 52 men) aged 19 to 71 years presenting for primary care to two neighborhood health centers between October 1994 and June 1995 were identified. Data collected included smoking status, alcohol use (CAGE questionnaire), depression (Vietnamese Depression Scale [VDS]), PPD status, stool ova and parasites, hepatitis B and syphilis serologies. RESULTS: Overall, 32% were smokers and significantly more men than women smoked (54% vs. 9%) (P < .00001). Although 24% of patients used alcohol, none responded positively to any of the CAGE questions. Using the VDS, 17% (17 of 99) were depressed; age 40 and older was the only sociodemographic factor associated with depression (P < .00001). Ova or parasites were found in 51% (41 of 80), and 63% of those infected (26 of 41) required treatment for pathogenic infections. Seventy percent (66 of 94) tested positive on the tuberculin skin test (PPD), and antituberculous medication was recommended in 39% (37 of 94). Eighty-three percent (80 of 96) had been exposed to hepatitis B, and 14% (13 of 96) were chronic hepatitis B carriers. CONCLUSIONS: Caring for special populations provides an opportunity to institute appropriate unique screening tests not recommended for the general population. In the case of new Vietnamese immigrants, routine screening protocols should include the following: testing for tuberculosis by PPD, stool ova and parasite examinations, hepatitis B serologies, and assessment for depression and smoking status. The CAGE questionnaire may not be an effective instrument for detecting alcohol abuse in this particular population. PMID- 9217640 TI - A community collaborative practice experience between Med/Peds and family practice. AB - PURPOSE: The medical literature has followed educational outcomes of Medicine Pediatric (Med/Peds) physicians, but limited studies exist as to clinical outcomes for these combined specialty physicians. Although a variety of practice environments are available for a growing number of Med/Peds physicians, a collaborative practice setting with family physicians may optimize the Med/Peds practice potential. This study investigates clinical practice outcomes and utilization efficiencies of collaborative Med/Peds family practice physicians within a community, which should provide an effective model in a growing managed care environment. PATIENTS AND METHODS: Two collaborative practice settings in a moderate size Midwest community were analyzed with respect to patient demographics and utilization scores provided by a practice management group and a nationally based health care network. Current Procedural Terminology (CPT) coding was used to follow demographic trends for over 45,000 patient visits for 1 year. Efficiency ratings (Z-scores) were used over the same year for over 6,000 health care network patient visits to 10 collaborative practice-based physicians, which were then compared to 141,101 community family practice patient encounters, 26,617 general internist patient encounters, and 29,995 patient encounters to pediatricians for utilization trends. RESULTS: Med/Peds and Family Practice patient care data reflected nearly identical patient demographics between specialties with only a few exceptions. Med/Peds physicians cared for three times the total number of children less than 2 years old. Med/Peds physicians experienced a higher complexity of illness, in part due to a 40% increase in internal referrals from family practice colleagues in the ambulatory care setting, while maintaining a third of the proportion of outpatient referrals. Cost-effective interoffice utilization was still maintained, supported by a more optimal efficiency rating for Med/Peds physicians compared to collaborative family practice colleagues. Inpatient efficiency was demonstrated for Med/Peds specialists even though a threefold increase in hospitalizations was observed, in part resulting from physicians within these collaborative practices arranging all newborn nursery and pediatric admissions be covered by Med/Peds physicians. Both collaborative primary care specialists demonstrated more cost-effective overall practice utilization scores when compared to community-based primary care specialists. CONCLUSIONS: Med/Peds physicians in this study have been trained to provide cost-effective patient care in both outpatient and inpatient settings. Decreased outside referrals by collaborative family practice physicians through utilization of Med/Peds colleagues serves to optimize practice economy by eliminating the threat of competition that exists among community-based generalists. Such a model helps to control an overused referral system to subspecialists. PMID- 9217641 TI - Clinical significance of eosinophilia in HIV-infected individuals. AB - PURPOSE: To assess the clinical significance of peripheral eosinophilia in HIV infected individuals. METHODS: In a retrospective case-control study we compared 42 HIV-infected patients (cases) with peripheral eosinophilia (absolute eosinophil count > 500 cells/mm3) with 84 HIV-infected controls without eosinophilia. Cases were matched to controls by date, and by CD4 cell count. Data on clinical parameters possibly associated with eosinophilia were collected and compared among cases and controls. RESULTS: Eosinophilia was seen in patients with late-stage HIV disease (median CD4 cell count of 26 cells/mm3). Cases were more likely to be black (52% versus 18%, P = 0.0001), have pruritus (50% versus 20%, P = 0.002), and have a physician-documented rash (76% versus 52%, P = 0.02). Specific cutaneous diagnoses that were more prevalent in cases versus controls were eosinophilic folliculitis (24% versus 1% P = 0.0001), atopic dermatitis (14% versus 1%, P = 0.01), and prurigo nodularis (7% versus O, P = 0.07). Other parameters commonly associated with eosinophilia such as allergic reactions, parasitic infection, malignancy, and adrenal insufficiency were not found at higher incidence in cases. CONCLUSIONS: Eosinophilia in AIDS patients is associated with cutaneous disease, but not with other conditions commonly associated with eosinophilia including parasitic infections, allergic reactions, or malignancy. Extensive work up for asymptomatic eosinophilia in patients with AIDS and cutaneous disease is not warranted. PMID- 9217642 TI - Lithium treatment in amiodarone-induced thyrotoxicosis. AB - PURPOSE: Amiodarone hydrochloride is an iodine-rich drug effective in the control of various tachyarrhythmias. It is known to cause refractory to thyrotoxicosis, which usually does not respond to regular antithyroid drugs. Lithium bicarbonate is a medication used to treat psychiatric disorders; it also influences thyroid production and release of hormones. We tried it in combination with propylthiouracil (PTU) for the treatment of amiodarone-induced thyrotoxicosis. PATIENTS AND METHODS: Twenty-one patients were studied. The first group (n = 5) was treated by amiodarone withdrawal only. The second group (n = 7) received PTU (300 to 600 mg), and the third (n = 9) PTU (300 mg) and lithium (900 to 1350 mg) daily. Patient selection was not randomized. The PTU + lithium group had more severe symptoms and signs of thyrotoxicosis, as well as thyroxine levels at least 50% above the upper limit of normal. They also had been on a longer course of amiodarone treatment (34.3 +/- 11.9 months) than the PTU-only (11.4 +/- 7.5) and the no-treatment (7.8 +/- 4.2) groups. RESULTS: While there was no difference between the first two groups in time until recovery (10.6 +/- 4.0 versus 11.6 +/- 0.5 weeks, respectively), the group receiving lithium normalized their thyroid function tests in only 4.3 +/- 0.5 weeks (P < 0.01 versus both other groups). T3 levels normalized even earlier-by 3 weeks of lithium treatment. No adverse effects of lithium were encountered, and the medication was stopped 4 to 6 weeks after achieving a normal clinical and biochemical state. CONCLUSIONS: We conclude that lithium is a useful and safe medication for treatment of iodine-induced thyrotoxicosis caused by amiodarone. We would reserve this treatment for severe cases only. Further studies are needed to find out whether in patients with this troublesome complication lithium therapy could permit continuation of amiodarone treatment. PMID- 9217643 TI - Inflammatory myocardial diseases and cardiomyopathies. AB - Inflammatory myocardial disease has been associated with a variety of infectious and noninfectious etiologies. It is associated with the development of dilated cardiomyopathy in some patients. Given its imprecise diagnosis, varied clinical presentation and undefined natural history, it is quite difficult to make broad generalizations regarding its evaluation and treatment. It is hoped continued application of new molecular biological and other techniques will shed further light on the pathophysiologic mechanisms of myocarditis in humans, thus pointing to therapeutic interventions. PMID- 9217644 TI - Developing strategic plans for academic general internal medicine. AB - There is an increasing need to train more primary care physicians. Therefore, it would be advantageous for academic general internal medicine (GIM) to develop strategies to meet these demands. Our GIM division developed a strategic planning process with the participant groups being the division faculty, a pertinent literature review (the surrogate expert), and selected medical administrators. The IDEALS systems design provided the conceptual framework for the strategic planning process. This process used the Delphi technique to develop the theoretically ideal work system based on the division's vision for its future role, and the Nominal Group Process Technique to create the recommended work system, using the Delphi results. The strategic planning process was economical and division faculty agreed that it was useful. PMID- 9217645 TI - Metabolic complications of urinary diversions: an overview. AB - Patients with urinary diversions present unique challenges to internists who have an important role in their long-term management. Advances in surgical techniques over the past 30 years have given rise to a number of urinary diversion procedures that use various intestinal segments. In its normal function, the intestine absorbs water and solutes. When placed in contact with the urinary stream, the intestine can create numerous metabolic abnormalities. These include bone disease, hepatobiliary disease, infection, malignancy, neurologic complications, nutritional deficiencies, and a number of electrolyte and acid base disorders. An overview of these metabolic abnormalities and their causes is provided, as well as recommendations for screening and management of patients. PMID- 9217646 TI - A patient with multiple myeloma presenting with severe polyneuropathy caused by necrotizing vasculitis. PMID- 9217647 TI - Survival after metformin-associated lactic acidosis in peritoneal dialysis- dependent renal failure. PMID- 9217648 TI - Subdural hematoma following roller coaster ride while anticoagulated. PMID- 9217649 TI - Esophageal stricture associated with alendronate therapy. PMID- 9217650 TI - Doctor Lively Day. PMID- 9217651 TI - Cardiomyopathy, nephropathy, and death in a 44-year-old man. PMID- 9217652 TI - Atrial fibrillation: prospective role of calcium channel blockers in open heart surgery. PMID- 9217653 TI - Yield of laboratory tests for case-finding in the ambulatory general medical examination. PMID- 9217654 TI - Yield of laboratory tests for case-finding in the ambulatory general medical examination. PMID- 9217655 TI - Perspectives on human papillomavirus infection. PMID- 9217656 TI - Epidemiology of genital human papillomavirus infection. AB - Although it is difficult to estimate the overall prevalence of genital human papillomavirus (HPV) infection, current figures suggest that visible genital warts are present in approximately 1% of sexually active adults in the United States and that at least 15% have subclinical infection, as detected by HPV DNA assays. Genital HPV infection is thus extremely common. The highest rates of genital HPV infection are found in adults 18-28 years of age. Although risk factors for infection are difficult to assess because of the high frequency of subclinical infection, it is clear that major risk factors for acquiring genital HPV infection involve sexual behavior, particularly multiple sex partners. Other possible risk factors for acquisition of genital HPV infection include oral contraceptive use, pregnancy, and impairment of cell-mediated immunity. Strong epidemiologic and molecular data link HPV infection to cervical and other anogenital cancers. The types of HPV most commonly detected in cancers are HPV-16 and HPV-18. In summary, genital HPV infection is common among sexually active populations and causes both benign and malignant neoplasms of the genital tract. PMID- 9217657 TI - Human papillomavirus and human disease. AB - Human papillomaviruses (HPVs) are associated with a spectrum of different diseases in humans, including common warts and genital warts. Of more serious concern is the connection between certain HPV types and some malignancies, particularly cervical and anal cancer. DNA from HPV-16 and HPV-18, two types frequently found in cervical cancer tissue, can immortalize cells in laboratory cultures, unlike DNA from HPV types associated with benign genital lesions. Although it is unclear how high-risk HPV types cause cancer, studies indicate that malignant transformation involves the viral E6 and E7 gene products, which may exert their effect by interfering with the cellular proteins that regulate cell growth. The vast majority of those infected do not develop malignancies, indicating that HPV infection alone is not enough to cause cancer. Cofactors such as cigarette smoking, may be required before neoplasia can occur. The potential seriousness of HPV infections is suggested by the observations that the number of genital HPV infections diagnosed is increasing and that cervical cancer is the second leading cause of cancer deaths in women throughout the world. PMID- 9217658 TI - Clinical presentation and natural course of anogenital warts. AB - The natural history of human papillomavirus (HPV) genital tract lesions is complex, partly because infection can appear in several forms and often follows a fluctuating course. The primary mode of transmission of genital strains of HPV is through sexual contact. Most clinically apparent genital warts are caused by HPV type 6 or 11. The main manifestations of anogenital warts are cauliflower-like condylomata acuminata that usually involve moist surfaces; keratotic and smooth papular warts, usually on dry surfaces; and subclinical "flat" warts, which can be found on any mucosal or cutaneous surface. Latent infections that can be detected only by the presence of HPV DNA, with neither macroscopic nor histologic abnormality, are probably the most common form of anogenital HPV infection, regardless of HPV type. Most untreated genital tract lesions eventually resolve spontaneously, but it is likely that latent or subclinical infection persists indefinitely. The natural history of anogenital HPV infection is likely influenced by the cell-mediated immune system. PMID- 9217659 TI - Diagnosis of human papillomavirus genital tract infection. AB - Although visible anogenital lesions are present in some persons infected with human papillomavirus (HPV), the majority of individuals with HPV genital tract infection do not have clinically apparent disease. Conventional viral detection assays, including serologic assays and growth in cell culture, are not available for the diagnosis and tracking of HPV infection. Papanicolaou tests are a valuable screening tool, but they miss a large proportion of HPV-infected persons. Accordingly, HPV DNA detection assays have become a key research tool in the detection of HPV infection, particularly in asymptomatic individuals. Several types of HPV DNA tests are now available, including Southern blots, dot blots, in situ hybridization, polymerase chain reaction, and solution hybridization (Hybrid Capture assay). Of these, the polymerase chain reaction assay is the most sensitive, whereas dot blots and solution hybridization are the least labor intensive. HPV DNA detection assays are not routinely used in screening patients, in part because the clinical relevance of asymptomatic infection is unclear. Nevertheless, these tests may be beneficial in confirming differential diagnoses and in providing prognostic information, particularly with respect to the HPV type involved. PMID- 9217661 TI - Counseling patients with genital warts. AB - Counseling patients about any sexually transmitted disease (STD) is difficult, for both the physician and the patient, but a diagnosis of genital warts presents particular challenges. For many patients, being told that they have any STD comes as a shock. Although fear is a common reaction, the relationship between human papillomavirus (HPV) and cancer has made the presence of genital warts especially frightening. This fear is heightened by the fact that treatment will not eradicate the underlying HPV infection, and the threat of recurring warts provides a constant reminder that the patient may never be truly cured. Thus a diagnosis of HPV involves many difficult issues, including poorly understood psychological sequelae in the patient, discomfort on the part of the physician, and notification of the patient's partner(s). Finally, issues of communication, lifestyle modification, and long-term management must be addressed. PMID- 9217660 TI - Therapeutic approaches to genital warts. AB - Although many treatments are available for genital warts caused by human papillomavirus (HPV), none are uniformly successful in the treatment of this disease. Most current treatment options work by destroying affected tissue, either by a cytotoxic or a physically ablative mode of action. Interferons have antiviral, antiproliferative, and immunomodulatory activities, but these have not translated into a high level of cure rates against warts. With all current treatments, recurrent warts are common. Therapies currently being investigated include a 5-fluorouracil/epinephrine collagen gel that achieves high concentrations of 5-fluorouracil at the site of injection. Other new treatment modalities focus on activating the host's immune system or improving the delivery of therapeutic compounds to the affected site. Imiquimod, a novel immune-response modifier, induces interferon and a number of other endogenous cytokines. A cream formulation containing 5% imiquimod resulted in good total clearance rates and generally tolerable side effects in controlled clinical trials of patients with external genital warts. Perhaps the most effective means for managing HPV disease would be a vaccine that prevents the occurrence of genital warts. Although it is unlikely that such a vaccine will be introduced in the near future, preliminary studies indicate that it may be possible to develop suitable prophylactic and therapeutic vaccines. PMID- 9217662 TI - What internists need to know: core competencies in women's health. ABIM Subcommittee on Clinical Competence in Women's Health. PMID- 9217664 TI - Giant cell arteritis in polymyalgia rheumatica. PMID- 9217663 TI - The Med/Peds physician in contemporary medical practice. PMID- 9217665 TI - High plasma levels of the soluble form of CD30 activation molecule reflect disease activity in patients with Wegener's granulomatosis. AB - PURPOSE: To determine the plasma levels of soluble CD30 (sCD30) in Wegener's granulomatosis (WG) patients, and to investigate the possible correlation of sCD30 with disease extent and activity. PATIENTS AND METHODS: sCD30 was determined by radioimmunoassay in 57 WG patients, 25 patients with rheumatoid arthritis (RA), 23 patients with bacterial infections and 21 healthy controls (HC). The extent and activity of WG disease were assayed according to disease extent index (DEI) and standard laboratory parameters. RESULTS: Plasma sCD30 levels in generalized WG (22.5 +/- 1.5 U/mL), but not in initial phase WG (12.1 +/- 4.0 U/mL), were significantly increased compared with HC (8.8 +/- 0.9 U/mL, P < 0.0001). Furthermore, of 11 generalized WG patients who received long-term follow-up, sCD30 levels declined when the disease activity changed from active disease to remission (29.1 +/- 1.9 U/mL to 15.9 +/- 1.8 U/mL, P = 0.0001). Similar results were observed in the whole group of generalized WG, eg, sCD30 levels in active disease (29.4 +/- 1.4 U/mL) were significantly higher than in partial remission (17.9 +/- 1.9 U/mL, P < 0.001) and in complete remission (13.7 +/- 3.3 U/mL, P < 0.001). No significant difference was noted between complete remission and HC. In addition, sCD30 levels were correlated with other parameters of disease extent and activity such as DEI, plasma levels of sIL-2R, PR3-ANCA, ESR and CRP. The sCD30 levels were increased in RA patients compared with HC (15.2 +/- 2.1 U/mL, P < 0.05), but no correlation was found between disease activity parameters and sCD30 levels. In contrast, in patients with bacterial infections sCD30 levels (6.9 +/- 0.9 U/mL) were not significantly different compared with HC. CONCLUSION: Plasma levels of sCD30 are not only significantly increased but also correlate with disease extent and activity in generalized WG. These findings suggest that sCD30 can act as a useful marker for evaluation of disease extent and activity, and that generalized WG may be associated with Th2 type immune response. PMID- 9217666 TI - Gender differences in patient preferences may underlie differential utilization of elective surgery. AB - PURPOSE: To study gender-specific preferences regarding timing of elective total joint replacement (TJR) surgery in patients with moderately severe osteoarthritis (OA) of the hip or knee. PATIENTS AND METHODS: Focus group discussions regarding TJR surgery were conducted among 18 women and among 12 men with moderately severe OA of the hip or knee. Discussions were tape recorded, transcribed, coded for themes, and evaluated semiquantitatively and qualitatively for gender differences. RESULTS: In general, men were more likely to choose surgery earlier in the disease than women and had higher expectations for surgical success. Women were more fearful of surgery. Women preferred to suffer arthritis pain rather than risk surgery, and indicated they would delay surgery to await better technology and to avoid disrupting caregiving roles for dependent spouses and others. CONCLUSION: Men and women differ in their willingness to accept continued functional decline, risks of surgery, and disruption of usual role. Gender differences may influence decisions regarding utilization of TJR. PMID- 9217667 TI - Microalbuminuria in patients with non-insulin-dependent diabetes mellitus relates to nocturnal systolic blood pressure. AB - PURPOSE: Microalbuminuria predicts early mortality in non-insulin-dependent diabetes mellitus patients (NIDDM). Our objective in the present study was to compare and assess the relationship between 24-hour, day and nocturnal ambulatory blood pressure (BP) and urinary albumin excretion rate (UAE) in microalbuminuric and normoalbuminuric NIDDM and in normal control subjects. PATIENTS AND METHODS: In the present cross-sectional study, 24 hour ambulatory BP (daytime BP and nocturnal BP) and HbA1c were compared in microalbuminuric (n = 10) and nonmicroalbuminuric NIDDM patients (n = 10) and in nondiabetic controls (n = 9). None of the patients were taking antihypertensive agents. RESULTS: In the microlbuminuric group, whereas 24 hour and daytime systolic BP differed significantly from control values (P < 0.025 and P < 0.05 respectively), there was no difference between diabetic groups. However, nocturnal systolic BP in the microalbuminuric group was significantly higher than in the normoalbuminuric diabetic patients (139 vs. 125) (P < 0.05) and a significant difference was also found between the NIDDM patients and the control group (139, 125 vs. 114) (P < 0.025). In multiple regression analysis, only nocturnal systolic BP showed a significant relationship with UAE (P < 0.05). CONCLUSIONS: We suggest that the higher nocturnal systolic blood pressure seen in our microalbuminuric NIDDM patients may contribute to the increased morbidity in this group. PMID- 9217668 TI - Postmenopausal estrogen replacement and tooth retention. AB - PURPOSE: To determine if estrogen replacement therapy (ERT) is associated with improved tooth retention and lower risk of edentulism (no natural teeth remaining) in a cohort of elderly women. PATIENTS AND METHODS: Subjects were 488 women, aged 72 to 95, who participated in the 23rd examination cycle (1994 to 1995) of the Framingham Heart Study, a population-based study begun in 1948. The number of teeth remaining and their location were recorded by a trained observer. History and duration of ERT were obtained from records kept since cycle 10 (1960 to 1963). Third molars were excluded from all analyses. RESULTS: Women who ever used ERT were younger than nonusers by 1 year (80 +/- 4 years, n = 184, versus 81 +/- 4 years, n = 304, P = 0.019). Estrogen users had more teeth remaining than nonusers (12.5 +/- 0.8 versus 10.7 +/- 0.8 versus 10.7 +/- 0.6 teeth, P = 0.046, mean +/- SE) after controlling for age, smoking status, and education. Duration of estrogen use was an independent predictor of the number of teeth remaining (P = 0.015) such that each 4.2-year interval of use was associated with an increased mean retention of 1 tooth. Long-term estrogen users (more than 8 years, n = 48) had an average of 3.6 more teeth than women who never used estrogen (14.3 +/- 1.5 versus 10.7 +/- 0.6 teeth, P < 0.02). The association with duration of use was present among different types of teeth (incisors, canines, and premolars) but less strong for molars. The odds of being edentulous were reduced by 6% for each 1-year increase in duration of estrogen use (odds ratio = 0.94, P = 0.038, 95% confidence interval = 0.90 to 0.99). CONCLUSIONS: These data suggest that ERT protects against tooth loss and reduces the risk of edentulism. The associations of estrogen use and tooth retention are evident for all but the molars. PMID- 9217669 TI - Optimal dietary calcium intake in primary hyperparathyroidism. AB - PURPOSE: The purpose of this study was to investigate whether dietary calcium intake in primary hyperparathyroidism is associated with differences in bone mineral density and biochemical parameters, and to determine whether these observations are related to 1,25-dihydroxyvitamin D. PATIENTS AND METHODS: Dietary calcium intake was determined from diet records in 71 unselected patients enrolled in an ongoing longitudinal study on the natural history of primary hyperparathyroidism. Subjects were placed into one of three dietary calcium groups based on their mean dietary calcium intake: very low (< 300 mg/day; mean = 199 +/- 14), low (300 to 800 mg/day; mean = 529 +/- 21), and US RDA (> 800 mg/day; mean = 1023 +/- 73). Biochemical indices were indicative of patients with modern day primary hyperparathyroidism, showing mild hypercalcemia (2.79 +/- 0.02 mmol/L), low normal serum phosphorus (0.90 +/- 0.03 mmol/L), elevated parathyroid hormone levels by midmolecule (764 +/- 69 pg/mL) and immunoradiometric (118 +/- 8 pg/mL) assays, and high normal 1,25-dihydroxyvitamin D (60 +/- 3 pg/mL) and urinary calcium excretion (6.3 +/- 0.4 mmol/day). Bone mineral density was measured by dual energy x-ray absorptiometry at the lumbar spine, right femoral neck and distal third of the nondominant radius for each subject. RESULTS: Over the entire range, there was no significant effect of dietary calcium on serum parathyroid hormone levels, calcium, phosphorus, 25-hydroxyvitamin D, 1,25 dihydroxyvitamin D, urinary calcium excretion, or bone mineral density of the lumbar spine, femoral neck or distal one-third radius. Serum 1,25 dihydroxyvitamin D was elevated in 37 patients (52%). Despite similarly low dietary calcium intake among patients with normal and elevated levels of 1,25 dihydroxyvitamin D (477 +/- 50 mg/day vs. 533 +/- 40 mg/day), patients with elevated levels of 1,25-dihydroxyvitamin D had higher parathyroid hormone levels by immunoradiometric assay (136 +/- 11 pg/mL vs. 97 +/- 10 pg/mL; P < .05), and urinary calcium (7.4 +/- 0.05 mmol/day vs. 5.1 +/- 0.05 mmol/day; P < .05 or 0.82 +/- 0.04 mmol/mmol creatinine vs. 0.56 +/- 0.04 mmol/mmol creatinine; P < .01). CONCLUSIONS: The data suggest that patients with normal levels of 1,25 dihydroxyvitamin D can liberalize their calcium intake without adverse consequences. However those with elevated levels of 1,25-dihydroxyvitamin D are advised to be more restrictive in order to prevent hypercalciuria. PMID- 9217670 TI - Immunochemical fecal occult blood test is not suitable for diagnosis of hemorrhoids. AB - PURPOSE: This study was conducted to clarify the diagnostic value of an immunochemical fecal occult blood test for hemorrhoids. PATIENTS AND METHODS: In a case-control study, an immunochemical fecal occult blood test with a 2-day method was carried out on 82 subjects with hemorrhoids, on 82 subjects with colorectal cancer, and on 82 healthy subjects. In a population-based cross sectional study, 29,714 subjects who received an immunochemical occult blood screening with a 2-day method were divided into two groups, according to the results of a questionnaire on hemorrhoids, and the positivity rate of an immunochemical test as well as the predictive value for colorectal cancer were compared in the two groups. Moreover, both an immunochemical occult blood test with a 2-day method and colonoscopy were conducted at the same time on asymptomatic subjects during a medical checkup. RESULTS: In the case-control study, the test was positive in 13.4% subjects with hemorrhoids, in 84.1% subjects with colorectal cancer, and in 4.9% healthy subjects, respectively, showing a significant difference in the detection rate between the two diseases (P < 0.001). In the population screening program, the test was positive in 6.9% subjects with hemorrhoids and in 6.5% subjects without hemorrhoids, and the predictive value was 3.2% in subjects with and without hemorrhoids, respectively, indicating no significant difference in the positivity rate as well as the predictive value between the two groups. Among 232 subjects in a medical checkup, 28 patients with hemorrhoids and 21 patients with colorectal polyp 1 cm or larger were diagnosed by colonoscopy, and the occult blood test was positive in 16.7% patients with hemorrhoids and in 52.4% patients with colorectal polyp, respectively. There was a significant difference in the sensitivity between the two disease groups (P < 0.05). CONCLUSIONS: These findings indicate that the immunochemical fecal occult blood is unsuitable for the diagnosis of the patients with hemorrhoids and an examination of the colorectum is necessary in cases where the occult blood test is positive but there is a sign of hemorrhoids. PMID- 9217671 TI - The diagnosis and management of fluid collections associated with pancreatitis. AB - Pancreatitis may be acute or chronic, mild or severe. Acute necrotizing pancreatitis remains the most serious form of acute pancreatitis and accounts for the majority of complications. Although there is an established nomenclature for pancreatitis and pancreatic fluid collections, such as pancreatic pseudocysts, it is not widely understood or recognized by physicians, including gastroenterologists. Because nonspecialists will be increasingly called upon to treat and appropriately refer patients with pancreatitis and its complications for more specialized care, it is important to understand the evolving treatment options for managing these patients. This article addresses and summarizes pancreatitis and its complications, particularly pancreatic collections. PMID- 9217672 TI - National survey of Clerkship Directors in Internal Medicine on the competencies that should be addressed in the medicine core clerkship. AB - PURPOSE: To prioritize competencies that should be addressed in the medicine core clerkship, assess factors influencing this prioritization, and estimate the percentage of clerkship time that should be devoted to inpatient versus outpatient care. METHODS: A national survey of the Clerkship Directors in Internal Medicine (CDIM) was used. Using explicit criteria, respondents assigned priority scores, on a 1 to 5 scale, to 17 general competencies and 60 disease specific clinical competencies pertinent to care of adult patients in inpatient. ambulatory, intensive care, and emergency settings. RESULTS: Ninety-three (75%) of 124 CDIM members responded. The highest mean priority scores were assigned to 6 general competencies: case presentation skills (4.65), diagnostic decision making (4.64), history and physical diagnosis (4.61), test interpretation (4.47), communication with patients (4.35), and therapeutic decision-making (4.12). Disease-specific clinical competency areas receiving the highest mean priority scores were: hypertension (4.57), coronary disease (4.53), diabetes mellitus (4.45), heart failure (4.42), pneumonia (4.39), chronic obstructive pulmonary disease (4.26), acid-base/electrolyte disorders (4.19), and acute chest pain (4.08). Priorities for general competencies were moderately correlated with importance to the practice of general internists (mean Spearman rho 0.49) and with importance to students pursuing careers outside internal medicine (mean Spearman rho 0.45), but only weakly correlated with the adequacy with which a competency was addressed in other parts of the curriculum. Respondents' mean recommended allocation of clerkship time was: 52% inpatient, 33% ambulatory care, 8% intensive care, and 7% emergency medicine. This time allocation did not differ by any characteristics of respondents. CONCLUSION: There is consensus among medicine clerkship directors that the medicine core clerkship should emphasize fundamental competencies and devote at least one third of the time to clinical competencies pertinent to ambulatory care. PMID- 9217674 TI - Doctor Weed in Montrose. PMID- 9217673 TI - The secondary prevention of coronary artery disease. AB - Randomized clinical trials demonstrate the efficacy of medical secondary prevention in coronary disease patients. The magnitude of risk reduction with exercise, diet, lipid modification, and smoking cessation is similar to other medical therapies for coronary disease such as aspirin, beta blockers, as well as coronary bypass surgery, (Table VI) In contrast to these therapies, however, secondary prevention stabilizes angiographic progression in about 50% of patients and induces regression in about 25% of patients. Both symptoms and perceived quality of life also are beneficially altered by secondary prevention programs, although possibly not by the magnitude reported for bypass surgery. These clinical trial results have led the American Heart Association, and the American College of Cardiology to strongly endorse secondary prevention. A reasonable projection based on these clinical trial data is that widespread use of these recommendations in the 12 million established coronary disease patients would significantly reduce coronary mortality and morbidity. PMID- 9217675 TI - Intravenous lidocaine for treatment-resistant pruritus. PMID- 9217676 TI - Thoracic complications following snake envenomization. PMID- 9217677 TI - Limbic encephalitis and small cell lung cancer. Clinical and immunological features. AB - Paraneoplastic limbic encephalitis (LE) is considered a particular manifestation of paraneoplastic encephalomyelitis (PEM), a remote effect of cancer almost always associated with anti-neuronal antibodies (anti-Hu; also called ANNA 1) and small cell lung carcinoma (SCLC). In order to define the frequency of anti-Hu antibodies in LE with SCLC and to analyse possible clinical differences between patients with and without anti-Hu antibodies, the charts of 16 patients with LE and SCLC were reviewed. Eight patients (50%) had anti-Hu antibodies (anti-Hu+) whereas eight patients (50%) had no detectable anti-neuronal antibodies (anti-Hu ). The clinical and laboratory features of LE and time to diagnosis of SCLC were similar in the anti-Hu+ and anti-Hu- groups. Involvement of other areas of the nervous system compatible with the diagnosis of PEM was observed in seven (87.5%) patients of the anti-Hu+ group but in only one (12.5%) of the anti-Hu- group (P = 0.012). Five patients, including four of the anti-Hu- group, had a partial improvement of the LE after treatment of the SCLC. Another anti-Hu- patient improved spontaneously. Six patients of the anti-Hu+ group died from the neurological disorder, whereas in the anti-Hu- group the cause of death was progression of the SCLC in the three patients who died. The results of this study indicate that the absence of anti-Hu antibodies does not rule out the presence of an underlying SCLC in patients with a clinical diagnosis of LE. Patients with LE and SCLC who are without anti-Hu antibodies are less likely to develop PEM and seem to improve more often after treatment of the cancer than those who present anti-Hu antibodies. PMID- 9217678 TI - Cytotoxic mechanisms in inflammatory myopathies. Co-expression of Fas and protective Bcl-2 in muscle fibres and inflammatory cells. AB - Expression of the Fas 'death receptor', Fas (CD95/APO-1) renders cells susceptible to programmed cell death ('apoptosis'), whereas Bcl-2 protects cells from apoptosis. Using fluorescence immunohistochemistry, we analysed Fas and Bcl 2 expression in muscle from five patients with polymyositis (PM), four patients with inclusion body myositis (IBM), three patients with dermatomyositis (DM), three patients with Duchenne muscular dystrophy (DMD) and three nonmyopathic controls. Fas (CD95) and Bcl-2 were not detected in control muscle, but expressed in muscle fibres and inflammatory cells in PM, IBM, DM and DMD. The proportion of Fas+ muscle fibres ranged from < 1 to 50%, and was higher in PM and IBM than in DM and DMD. On average, the Fas+ muscle fibres were smaller (median diameter, 10 microns; range, 7-32 microns) than the Fas- fibres (median, 36 microns; range, 10 60 microns). Less than 10% of the Fas+ muscle fibres co-expressed the regeneration marker CD56 (neural cell adhesion molecule N-CAM). In PM and IBM, the proportion of Fas+ muscle fibres was higher among fibres invaded or contacted by T cells than among fibres not contacted by T cells (P < 0.01). The proportion of Fas+ fibres co-expressing Bcl-2 was 76 +/- 16% in PM, 100% in IBM and 63 +/- 23% in DM. Fas and Bcl-2 expression was also noted in inflammatory cells in PM, IBM, DM and DMD. Using the terminal deoxytransferase-catalysed DNA nick end labelling technique for detection of nuclear DNA fragmentation, none of myonuclei, and < 0.1% of inflammatory cell nuclei, showed signs of apoptosis. Our results suggest that, although Fas expression confers susceptibility to Fas mediated apoptosis, Fas-expressing muscle fibres and inflammatory cells are protected by the anti-apoptotic protein Bcl-2. PMID- 9217679 TI - Ring chromosome 20 and nonconvulsive status epilepticus. A new epileptic syndrome. AB - Six cases of epilepsy associated with ring chromosome 20 are presented. The study of these cases and 20 cases reported in the literature revealed that they constitute a distinct epileptic syndrome: frequent seizures consisting of a prolonged confusional state, with or without additional motor seizures, and an ictal EEG pattern of long-lasting bilateral paroxysmal high-voltage slow waves with occasional spikes. Neurological examination results were normal, and neuroimaging studies often failed to disclose a brain lesion. The seizures were resistant to antiepileptic drug therapy. Comparison of the electroclinical features of nonconvulsive status epilepticus in six patients with and four patients without ring chromosome 20 revealed that the group with the chromosomal anomaly had more frequent, comparatively brief episodes of confusion associated with a less prominent spike component on the EEG. We propose that epilepsy associated with ring chromosome 20 constitutes a new syndrome that may provide an opportunity to scrutinize a genetic mechanism of epilepsy. PMID- 9217680 TI - Differences between hereditary motor and sensory neuropathy type 2 and chronic idiopathic axonal neuropathy. A clinical and electrophysiological study. AB - To evaluate whether chronic idiopathic axonal polyneuropathy (CIAP) should be considered as hereditary motor and sensory neuropathy type 2 (HMSN type 2), we compared the clinical features of 48 patients with CIAP with those of 47 patients with HMSN type 2. In addition, we studied electrophysiological data in 20 patients with CIAP and in 20 patients with HMSN type 2. We found, in patients with HMSN type 2, that the initial symptoms were predominantly motor and that weakness and handicap were more severe and skeletal deformities more frequent, compared with those of CIAP patients. Electrophysiologically, the tibialis anterior muscle showed more denervation in patients with HMSN type 2, consistent with the predominance of motor symptoms. There was no important effect of age of onset on clinical features in HMSN type 2 patients. We conclude that in an individual patient with a sensory or sensorimotor idiopathic axonal polyneuropathy and no family history of polyneuropathies, the diagnosis HMSN type 2 is unlikely. However, if motor symptoms predominate, the diagnosis of HMSN type 2 should be considered. PMID- 9217681 TI - Evidence for lateral premotor and parietal overactivity in Parkinson's disease during sequential and bimanual movements. A PET study. AB - Patients with Parkinson's disease have great difficulty in performing sequential and bimanual movements. We used H2(15)O PET to study the regional cerebral blood flow associated with performance of sequential finger movements made unimanually and bimanually in a group of Parkinson's disease patients and a group of control volunteers. In controls, sequential finger movements led to activation of the contralateral motor cortex and inferior parietal cortex (Brodmann area 40), the lateral premotor cortex and bilateral supplementary motor area. No prefrontal activation was seen. Sequential finger movements in the Parkinson's disease group were associated with a similar pattern of activation but there was relative impairment of activation in the mesial frontal and prefrontal areas. A novel finding was the presence of relative overactivity in the lateral premotor and inferolateral parietal regions. We conclude that in Parkinson's disease there is a switch from the use of striato-mesial frontal to parietal-lateral premotor circuits in order to facilitate performance of complex finger movements. PMID- 9217682 TI - Proprioceptive control of wrist movements in Parkinson's disease. Reduced muscle vibration-induced errors. AB - The effects upon the trajectories of practised slow (approximately 9 degrees/s) voluntary wrist-extension movements of applying vibration to the tendon of an antagonist muscle (flexor carpi radialis) during the course of the movement have been studied in patients with idiopathic Parkinson's disease and age-matched healthy individuals. In both patient and control groups, flexor vibration elicited undershooting of wrist-extension movements. Wrist extensor and flexor surface EMG recordings indicated that, in patients and controls, such undershooting resulted principally from sustained reductions in extensor (prime mover) activity. Small vibration reflexes were commonly elicited in the wrist flexors which, in both Parkinson's disease and healthy subjects, were usually otherwise virtually quiescent during these slow extension movements. The amplitudes of such vibration reflexes did not differ systematically between patient and control groups and appeared inadequate to have exerted an appreciable braking action upon the extension trajectories. However, the extent of vibration induced undershooting was, on average, significantly less in the Parkinson's disease group. In a subgroup of patients with asymmetrical parkinsonism the effects of antagonist vibration upon wrist movements of the more and less affected limb were compared. The degree of vibration-induced undershooting was significantly smaller on the more affected side. This finding suggests that disturbed proprioceptive guidance of voluntary movements in Parkinson's disease is related to the severity of clinical motor deficits. A small number Parkinson's disease patients were studied 'ON' and 'OFF' their routine anti-parkinsonian medication. A non-significant tendency was found for vibration-induced errors to be less marked in the 'OFF' state. In a separate series of experiments, under isometric conditions, vibration-induced EMG changes were recorded whilst subjects attempted to maintain a steady (15% maximum) voluntary wrist extensor effort. Results in control subjects suggested that prolonged flexor vibration produced significant tonic reflex reciprocal inhibition of the extensor muscles. However, the strength of reflex inhibition appeared sufficient to account for only a small fraction of the undershooting observed during the movement tasks. Thus, our results are consistent with the existence of an abnormality of higher-level proprioceptive integration in Parkinson's disease in which there is a mismatch of sensory (proprioceptive) and motor (corollary discharge) information. PMID- 9217683 TI - Involvement of spinal recurrent inhibition in spasticity. Further insight into the regulation of Renshaw cell activity. AB - Changes in the excitability of the soleus H-reflex are were studied after oral administration of L-acetylcarnitine, a cholinomimetic substance, in eight healthy control subjects and 23 spastic patients presenting with slowly progressive paraparesis (n = 10), a cord lesion (n = 9) and a cerebral lesion (n = 4). Changes in the amount of recurrent inhibition of soleus motor neurons at rest were also estimated in order to assess the level of activity of Renshaw cells before and after L-acetylcarnitine administration. Recurrent inhibition elicited by a conditioning reflex discharge (H1) was assessed by a subsequent test reflex (H'). Four patients lacked an H' reflex. In approximately 50% of the remaining patients, recurrent inhibition was normal, while in the other half there was evidence of reduced or absent inhibitory activity at rest. Pooling the data relative to the effect of L-acetylcarnitine on the H-reflex in relation to the strength of recurrent inhibition disclosed that the ratio of peak-to-peak amplitude values of the maximum H reflex to maximum M wave responses (Hmax:Mmax) was reduced in all the cases in which the recurrent inhibition at rest was normal, while such a reduction was never observed in the patients in whom recurrent inhibition was found to be decreased at rest. In the former cases, the size of the H' reflex evoked by the same conditioning H1 discharge was further depressed after L-acetylcarnitine, pointing to a potentiating effect of the drug on Renshaw cells; in the latter cases no such effect was seen. A significant decrease in the mean Hmax:Mmax ratio after L-acetylcarnitine intake was also seen in the healthy control subjects. Possible changes in the amount of presynaptic inhibition on Ia terminals on soleus motor neurons after L-acetylcarnitine were ruled out. It is proposed that the differential effect of the drug on the H reflex excitability is directly related to the level of Renshaw cell activity, a reduction of which probably follows a lesion interrupting reticulo-spinal pathways with tonic facilitatory influences on Renshaw cells. These findings support the hypothesis that Renshaw cell excitability is set via cortico-reticulo spinal systems. PMID- 9217684 TI - Effect of otolith dysfunction. Impairment of visual acuity during linear head motion in labyrinthine defective subjects. AB - Visual symptoms emerging after the loss of vestibular function are usually attributed to the dysfunction of semicircular canal vestibulo-ocular reflexes, as they have been shown to stabilize vision during angular head movements. However, natural head displacements involve both angular and linear motion, and therefore visual instability may occur because of defective otolith-ocular reflexes (OORs) which are the eye movements evoked by linear head acceleration. In this paper, the relationship between OORs and visual acuity during linear head motion was studied in normal subjects and 14 patients with bilateral loss of caloric responses. OORs were elicited in darkness by step acceleration (0.24 g) of the whole body along the interaural axis. Latency, slow phase velocity and asymmetry of the OOR were measured from the desaccaded and averaged electrooculographic trace. Visual acuity was assessed during sinusoidal lateral oscillation of the subject viewing an earth-fixed target, and vice versa with the subject stationary and the target moving at 0.5, 1.0 and 1.5 Hz. The task was to recognize numbers flashing up on a three digit light-emitting diode visual display. Normal subjects had symmetrical OORs with short latencies (< 130 ms). In patients, OORs were either absent (n = 2) or abnormal with asymmetries (n = 8), diminished velocities (n = 4) or prolonged latencies (n = 6). At high frequency oscillation (1.5 Hz), normal subjects invariably recognized more numbers during self-motion compared with target motion, whereas most patients did not. In patients, abnormal dynamic visual acuity was correlated with absent or delayed OOR responses. This is the first demonstration of a functional role of the OORs in that they contribute to visual stabilization during high frequency linear head motion. Bilateral vestibular failure commonly affects the OORs and thereby compromises dynamic visual acuity. PMID- 9217685 TI - Memory and executive functions in amnesic and non-amnesic patients with aneurysms of the anterior communicating artery. AB - Ruptured and repaired anterior communicating artery (ACoA) aneurysm can result in devastating impairments involving memory, executive function, confabulation and personality change. Importantly, traditional cerebral areas implicated in amnesia are not damaged, yet amnesia can still be manifested. While ACoA patients show normal visual-constructional skills (i.e. copy scores) on the Rey-Osterrieth complex figure test, recall is often impaired. What is unclear is whether impaired recall is attributable to problems in encoding, accelerated rates of forgetting, retrieval or some combination. To disentangle these issues, we examined 10 patients with ruptured aneurysms of the ACoA, using the Rey organizational and extended memory procedure which uses an organizational procedure for enhancing immediate recall and provides added sensitivity by combining recall with non-recall measures (e.g. recognition, spatial discrimination and spatial assembly). The major findings were: (i) immediate recall in amnesics was improved by providing an organizational strategy; (ii) following the organization trials, amnesics and non-amnesics retained information to a comparable extent over a 30-min delay; (iii) two subgroups of amnesics emerged, those subjects impaired in acquisition and a second group with impaired retrieval; (iv) all subjects showed preserved memory on non-recall measures. These findings have important implications with respect to using organizational strategies in cognitive treatments and in using non-recall measures in improving the validity and reliability of patient assessment. PMID- 9217686 TI - The temporal variant of frontotemporal dementia. AB - Frontotemporal dementia is a dementia syndrome with diverse clinical characteristics. Based upon clinical parameters and single photon emission computed tomography, we identified 47 frontotemporal dementia subjects. In 10 of these 47 the primary site of brain dysfunction was anterior temporal and orbital frontal with other frontal regions relatively spared. In this temporal lobe variant (TLV) of frontotemporal dementia, five of the subjects had more severe left-sided, and five had more right-sided, hypoperfusion. The clinical, neuropsychological and neuropsychiatric features of predominantly left-sided (LTLV) and right-sided (RTLV) TLV subjects are discussed and contrasted with more frontal presentations of frontotemporal dementia. In LTLV, aphasia was usually the first and most severe clinical abnormality RTLV patients presented with behavioural disorders characterized by irritability, impulsiveness, bizarre alterations in dress, limited and fixed ideas, decreased facial expression and increased visual alertness. These findings suggest that: (i) frontotemporal dementia is clinically heterogeneous with bitemporal and inferior frontal lobe dysfunction contributing to the clinical presentation; (ii) behavioural disturbance and aphasia are the most prominent features of predominantly temporal subtypes of frontotemporal dementia; (iii) the right and left anterior temporal regions may mediate different behavioural functions. The results of this study suggests that TLV offers a valuable source of information concerning the behavioural disorders seen with combined anterior temporal and inferior frontal lobe dysfunction. PMID- 9217687 TI - Differentiation of hypoglycaemia induced cognitive impairments. An electrophysiological approach. AB - Effects of hypoglycaemia on distinct cognitive processes were assessed with event related brain-potential (ERP) measures and reaction times in a hybrid selective attention/response-choice task. One group of subjects received a euglycaemia hypoglycaemia-euglycaemia treatment order and a second group a euglycaemia placebo(euglycaemia)-euglycaemia sequence of treatments. During hypoglycaemia, ERP measures of selective attention (selection negativity), response choice (lateralized readiness potential) and reaction time were delayed compared with baseline performance. After restoration of euglycaemia, the onset of the selection negativity returned to baseline, whereas the lateralized readiness potential was still delayed, and error frequencies remained elevated. These results suggest that hypoglycaemia delays both the stimulus selection and the motor-response selection. They further suggest that the stimulus-selection process recovers quickly after restoration of euglycaemia, but that the response selection process does not. Slow shifts in cortical potentials with a broad frontal distribution that occurred during hypoglycaemia, are discussed in relation to frontal lobe mechanisms involved in the control of subordinate, modality-specific selection mechanisms. PMID- 9217688 TI - The right brain hemisphere is dominant in human infants. AB - The development of functional brain asymmetry during childhood is confirmed by changes in cerebral blood flow measured at rest using dynamic single photon emission computed tomography. Between 1 and 3 years of age, the blood flow shows a right hemispheric predominance, mainly due to the activity in the posterior associative area. Asymmetry shifts to the left after 3 years. The subsequent time course of changes appear to follow the emergence of functions localized initially on the right, but later on the left hemisphere (i.e. visuospatial and later language abilities). These findings support the hypothesis that, in man, the right hemisphere develops its functions earlier than the left. PMID- 9217689 TI - The neuroendocrine axis in patients with multiple sclerosis. AB - We investigated the basal and dynamic regulation of the hypothalamo-pituitary adrenal (HPA), hypothalamo-pituitary-thyroid (HPT) and hypothalamo-pituitary gonadal axes and prolactin secretion in 52 patients with clinically definite multiple sclerosis. These patients also had gadolinium enhanced brain MRI scans and were divided into relapsing-remitting, secondary progressive and primary progressive subgroups. These subgroups were compared with healthy controls and a group of patients with other neurological diseases. The cortisol diurnal rhythm was preserved in all groups of patients. The time-integrated cortisol response to human corticotropin-releasing hormone (CRH) stimulation was lower in the patients with secondary progressive multiple sclerosis, compared with patients with primary progressive multiple sclerosis and healthy subjects. The time-integrated beta-endorphin response to CRH was greater in the patients with relapsing remitting multiple sclerosis compared with the others. Feedback regulation assessed by dexamethasone suppression was normal. Serum testosterone was low in 24% of male multiple sclerosis patients and oestradiol was low in 25% of pre menopausal female multiple sclerosis patients, whereas prolactin and the HPT function were normal. Correlations with C-reactive protein (CRP) and MRI suggest that activation of the HPA axis in multiple sclerosis patients is secondary to an active inflammatory stimulus. PMID- 9217690 TI - Impaired cortico-bulbar tract function in dysarthria due to hemispheric stroke. Functional testing using transcranial magnetic stimulation. AB - We investigated cortico-lingual and cortico-orofacial tract function utilizing transcranial magnetic stimulation in 18 consecutive patients with dysarthria due to hemispheric stroke. Delayed responses (conduction time > mean + 2.5 SD of that of 43 controls) or absent responses were considered abnormal. In all patients, motor-cortex stimulation of the lesion side demonstrated absent (13 patients) or delayed (five patients) responses to the tongue bilaterally (17 patients) or unilaterally (one patient). In 14 patients the contralateral orofacial responses were either absent (13 patients) or delayed (one patient). According to the electrophysiological findings, all lesions revealed by CT or MRI, were located within the pyramidal tract at the lower motor cortex (n = 4), the corona radiata (n = 7), and the genu of the internal capsule (n = 3) or its posterior limb (n = 4). We conclude that interruption of the cortico-bulbar tract fibres to muscles involved in articulation is a frequent cause of dysarthria in hemispheric stroke. PMID- 9217691 TI - Primary progressive multiple sclerosis. AB - Patients with multiple sclerosis who develop progressive disability from onset without relapses or remissions pose difficulties in diagnosis, monitoring of disease activity and treatment. There is a need to define the diagnostic criteria for this group more precisely and, in particular, to describe a comprehensive battery of investigations to exclude other conditions. The mechanisms underlying the development of disability and the role of MRI in monitoring disease activity in this clinical subgroup require elucidation, particularly in view of the lack of change on conventional imaging in the presence of continuing clinical deterioration. The prognosis is poor and there are currently no treatment trials for this form of the disease. PMID- 9217692 TI - Predicting survival in the intensive care unit. PMID- 9217693 TI - Glatiramer acetate for relapsing multiple sclerosis. PMID- 9217694 TI - Tizanidine for spasticity. PMID- 9217695 TI - Topical butenafine for tinea pedis. PMID- 9217696 TI - Lymphocyte subset changes between 3 and 15 months of age in infants born to HIV seropositive women in South Africa. AB - The evolution of T-lymphocyte subsets during infancy in perinatally HIV-infected African babies has not been previously described. In a hospital-based cohort study, T-lymphocyte subset changes were investigated in 72 South African black children born to HIV seropositive mothers. Sixteen (22.2%; children were classified as infected and 56 (77.8%) as uninfected by 18 months of age. Four (25%) of the infected infants died before the age of 9 months from HIV-related disease. The CD4 and CD8 T-lymphocyte subsets, expressed in absolute numbers, as percentages, percentiles or as ratios, were clear indicators of HIV infection at all ages between 3 and 15 months. The most marked changes were a decreased percentage of CD4 cells and an increase in percentage of CD8 cells in the infected group. In the 4 infected infants who died, CD8 count and CD4:CD8 ratio clearly predicted poor clinical outcome at 3 months. Taken together, both CD4:CD8 ratio and CD4 percentage are reliable markers of HIV infection in an African paediatric population; however, a raised CD8 lymphocyte count rather than a CD4 count is a more specific prognostic marker of disease progression in HIV infected children. PMID- 9217697 TI - Urban epidemic of bubonic plague in Majunga, Madagascar: epidemiological aspects. AB - After an absence of 62 years, an epidemic of plague occurred in the harbour city of Majunga (Madagascar) from July 1995 to March 1996, following sporadic cases in March and May 1995. By 15 March 1996, 617 clinically suspected cases of bubonic plague had been notified. Laboratory testing was carried out for 394 individuals: 60 (15.2%) were confirmed to have bubonic plague and 48 (12.2%) were considered as presumptive cases. The incidence was significantly higher in males in all age groups and in both sexes in the 5-19 age group. Twenty-four deaths were related to plague, but early treatment with streptomycin has confirmed its effectiveness insofar as the case-farality ratio was only 8.7% among confirmed and presumptive cases admitted to hospital. The difficulty of clinically diagnosing bubonic plague was affirmed. The disease met favourable conditions through the poverty and low level of hygiene prevalent in most parts of Majunga. PMID- 9217698 TI - Increasing prevalence of penicillinase-producing Neisseria gonorrhoeae and the emergence of high-level, plasmid-mediated tetracycline resistance among gonococcal isolates in The Gambia. AB - One hundred and three strains of Neisseria gonorrhoeae isolated from a periurban STD clinic in The Gambia were studied for antimicrobial susceptibility, plasmid profile, and serogroup using standard procedures. Seventy-nine (77%) were penicillinase producers (PPNG) and fully resistant to penicillin (MIC > or = 8 mg/l). One isolate showed chromosomally induced resistance to penicillin (MIC 2 mg/l). None of the isolates was sensitive to tetracycline; 16 (16%) showed intermediate resistance (MICs 1-8 mg/l) and 87 (84%) showed high-level plasmid mediated resistance (TRNG) (MICs > 10 mg/l). This is the first report of TRNG in The Gambia. Only 6 (6%) strains were fully sensitive to trimethoprim sulphamethoxazole (MIC < 8 mg/l); 78 (76%) showed intermediate level resistance (MICs 8-16 mg/l) and 19 (18%) were fully resistant (MIC > 32 mg/l). Indications of an increase in MIC to ciprofloxacin and ceftriaxone were found in 6 (6%) and 1 (1%) strains, respectively, although all remained fully sensitive (MICs 0.004 0.03 mg/l and 0.001-0.015 mg/l). All PPNG and TRNG strains carried the 3.2 MDa and 25.2 MDa plasmids, respectively. All isolates carried the 2.6 MDa cryptic plasmid and 9 (3 PPNG and 6 non-PPNG) carried the 24.5 MDa conjugative plasmid. Forty-four (43%) strains were typed group W1, 58 (56%) W11/111 and 1 had cross reacting antigens. Because PPNG are frequently encountered and high-level TRNG is now prevalent, the newer cephalosporins and quinolones must now be considered as first-line drugs for the treatment of gonorrhoea in The Gambia. PMID- 9217699 TI - Diagnosis of visceral leishmaniasis by the polymerase chain reaction using blood, bone marrow and lymph node samples from patients from the Sudan. AB - We have evaluated the sensitivity of the polymerase chain reaction (PCR) as a diagnostic tool for Leishmania donovani using blood, bone marrow and lymph node samples from Sudanese patients with a confirmed infection. Forty patients were diagnosed by microscopic examination of bone marrow or lymph node samples. The PCR was able to detect parasite DNA in 37 out of 40 blood samples. In bone marrow and lymph node samples, the PCR was able to detect parasite DNA in all 7 and 6 samples, respectively. We suggest that the PCR should be considered as a valuable and sensitive tool for the diagnosis of L. donovani infection. However, if PCR diagnosis is to supplement or even replace microscopic diagnosis in developing countries, a large number of patients with no apparent signs of infection and patients with other diseases have to be tested in order to evaluate its true potential. PMID- 9217700 TI - Road traffic injuries in developing countries: a comprehensive review of epidemiological studies. AB - Motor vehicle accidents are the leading cause of death in adolescents and young adults worldwide. Nearly three-quarters of road deaths occur in developing countries and men comprise a mean 80% of casualties. This review summarizes studies on the epidemiology of motor vehicle accidents in developing countries and examines the evidence for association with alcohol. PMID- 9217701 TI - Efficacy of artemisinin and mefloquine combinations against Plasmodium falciparum. In vitro simulation of in vivo pharmacokinetics. AB - The activity of artemisinin in combination with mefloquine was tested in vitro against a chloroquine-sensitive (F32) strain of Plasmodium falciparum. A method of repetitive dosing and extending the culture observation period to 28-30 days was used to mimic the in vivo pharmacokinetic situation. Plasmodium falciparum was exposed to artemisinin from 10(-8) to 10(-5) M, mefloquine from 3 x 10(-9) to 10(-5) M and their combinations. The exposure time for artemisinin was 3 hours twice daily and for mefloquine 24 hours. The drug-dosing duration was 3 days. Neither artemisinin nor mefloquine alone provided radical clearance of P. falciparum, even when maximum concentrations (10(-5) M) were applied. The antiparasitic activity of artemisinin and mefloquine were significantly higher when dosed alone. Effective concentrations for different degrees of inhibition (EC 50, 90 and 99) of both artemisinin and mefloquine respectively were significantly lower when used in combination. At concentrations normally reached in vivo, this effect was clearly synergistic (P = 0.016) Our in vitro model of intermittent dosing of artemisinin and mefloquine combinations for 3 days provides significant evidence of positive interaction between the two compounds. Lower combination concentrations around the MIC-values for the individual compounds showed synergistic effect, and high concentrations showed additive effect. This indicates that such drug combinations may provide radical clearance at concentrations lower than those required for single-drug treatment. PMID- 9217702 TI - Molecular typing of HLA class I and class II antigens in Indian kala-azar patients. AB - HLA has been shown to be associated with many diseases. To find out whether host genetic factors like the HLA are involved in susceptibility to kala-azar (visceral leishmaniasis) in India, we formulated an association study with genetically related controls. All samples were typed by PCR SSOP (sequence specific oligonucleotide probes) for HLA class I (A and B) and class II (DR) antigens. The test of association we used was the transmission disequilibrium test (TDT). No significant evidence for association with any of the three HLA loci was obtained. PMID- 9217703 TI - Onchocerca volvulus: immunohistochemical and immunoelectron microscopical distribution of a polyamine oxidizing enzyme. AB - We studied the distribution of a polyamine oxidizing enzyme (PAO) in Onchocerca volvulus and other nematode parasites by immunohistochemistry and electron microscopy with immunogold technique using a polyclonal antiserum raised against purified PAO from Ascaris suum. In adult O. volvulus the protein was localized in the outer zone and the area of the basal labyrinth of the hypodermis and occasionally in the outer zone of the uterine epithelium. Further, the fluid in the body cavity was strongly stained. No specific labelling was observed in the cuticle, muscles, epithelia of intestine, ovaries, testis and vas deferens or in sperm, oocytes and embryos. Third-stage larvae of O. volvulus in Simulium soubrense showed strong staining; the same was observed in Anisakis sp. larvae, where the inner and outer zone of the hypodermis were strongly labelled. All mature, intact and dead microfilariae in nodules, skin and lymph nodes were well stained and it was possible to show that the cytoplasm of the hypodermal cells, but not the mitochondria, nuclei or other organelles of muscle cells, was preferentially labelled by immunogold particles. Investigation of adult A. suum presented specific labelling of the hypodermis, but the basal labyrinth was more strongly marked than the outer zone. PMID- 9217704 TI - Protective effect of Trypanosoma rangeli against infections with a highly virulent strain of Trypanosoma cruzi. AB - We investigated the protective effect of Trypanosoma rangeli against infection with Trypanosoma cruzi in animal models of various ages and with different doses of inoculum. The age of the mice and the dose of parasites determined the course of the infection. When T. cruzi was inoculated into mice after challenge with T. rangeli, parasitaemia was more controlled, mortality decreased and histopathology showed lower inflammatory infiltration and pseudocysts. This study proposes a new murine model of the protective effect of recombinant proteins of T. rangeli for possible application in the vaccines field. PMID- 9217705 TI - Malaria-related beliefs and behaviour in southern Ghana: implications for treatment, prevention and control. AB - A research infrastructure was established in two ecological zones in southern Ghana to study the variables of malaria transmission and provide information to support the country's Malaria Action Plan (MAP) launched in 1992. Residents' beliefs and practices about causes, recognition, treatment and prevention of malaria were explored in two ecological zones in southern Ghana using epidemiological and social research methods. In both communities females constituted more than 80% of caretakers of children 1-9 years and the illiteracy rate was high. Fever and malaria, which are locally called Asra or Atridi, were found to represent the same thing and are used interchangeably. Caretakers were well informed about the major symptoms of malaria, which correspond to the current clinical case definition of malaria. Knowledge about malaria transmission is, however, shrouded in many misconceptions. Though the human dwellings in the study communities conferred no real protection against mosquitoes, bednet usage was low while residents combatted the nuisance of mosquitoes with insecticide sprays, burning of coils and herbs, which they largely considered as temporary measures. Home treatment of malaria combining herbs and over-the-counter drugs and inadequate doses of chloroquine was widespread. There is a need for a strong educational component to be incorporated into the MAP to correct misconceptions about malaria transmission, appropriate treatment and protection of households. Malaria control policies should recognize the role of home treatment and drug shops in the management of malaria and incorporate them into existing control strategies. PMID- 9217706 TI - Acceptability and use of insecticide impregnated bednets in northern Ghana. AB - A district-wide study was undertaken in a rural population of northern Ghana to identify factors influencing the acceptance and use of insecticide-impregnated bednets (IIBNs). A series of focus group discussions were conducted during 2 years of implementation of IIBNs to gauge community reactions to the introduction of the nets and a structured questionnaire was administered to approximately 2000 randomly selected individuals. Although the IIBNs were accepted and used because they provided protection from mosquito bites, seasonal factors, patterns of use, and questions of cost were key factors likely to influence the dissemination and effectiveness of bednets. Use of the bednets was highly seasonal. Almost all recipients used their IIBNs in the rainy season (99%), corresponding to the period of high mosquito density and 20% used them in the dry seasons, the period of low mosquito density. Mothers with young children were more likely to wash the bednets frequently (because the children soiled the bednets with faeces and urine), resulting in no protection from the insecticide. Provision of wider bednets, or the provision of plastic sheets with the bednets or possible incorporation of the insecticide in washing soaps could improve protection for young children. The success of the promotion of IIBNs in malaria control programmes will depend on the cost of the package and the time of year that it is delivered. Financing mechanisms for individual and village groups are discussed. Social research effectively monitored the intervention in this study, and it should be included as an important component of national malaria control programmes. PMID- 9217707 TI - Tobacco: promises are not enough. PMID- 9217708 TI - Research into intensive-care admissions. PMID- 9217709 TI - Ivermectin where Loa loa is endemic. PMID- 9217710 TI - How to stop ACE-inhibitor-induced cough. PMID- 9217711 TI - Lancet ombudsman's first report. PMID- 9217712 TI - Mortality among appropriately referred patients refused admission to intensive care units. AB - BACKGROUND: The provision of intensive care is a perplexing issue for clinicians and the public. Concerns about the apparent lack of beds and the appropriateness of the patients admitted are tempered by the high cost of providing this service. As part of a study commissioned by the UK Department of Health, we tested the hypothesis that there is excess mortality among patients who are refused admission to intensive-care units. METHODS: All referrals to six intensive-care units with different numbers of beds were monitored during a 3-month period. Data on mortality 90 days after first referral were obtained from family physicians for all patients known to be alive at hospital discharge. We adjusted, where possible, for confounding, including for age, sex, appropriateness of referral, disease severity, surgery and emergency categories, and bed provision. We did multivariate analysis by multiple logistic regression to compare the adjusted 90 day mortality rates for patients who were refused admission and for those admitted. FINDINGS: 480 patients were admitted and 165 were refused admission. 90 days after referral there had been 178 (37%) deaths among the admitted group and 75 (46%) among the refused group. After multivariate adjustment, 113 patients appropriately referred for intensive care but refused admission to their first choice intensive-care unit had a relative risk of death of 1.6 (95% CI 1.0-2.5), compared with the group of appropriately admitted cases with medium APACHE II scores for disease severity. Age, the assessed need for treatment or monitoring interventions, and emergency status also contributed to differences in mortality among all referrals. Bed provision did not contribute significantly to excess mortality. INTERPRETATION: Although this study is observational and case-mix adjustment is incomplete, we found a higher rate of attributable mortality in patients who were refused intensive care, particularly for emergency cases. We question whether the provision of more beds alone would be a solution and conclude that there is an urgent need for more appropriate admission and discharge criteria. PMID- 9217713 TI - Retinopathy of prematurity in middle-income countries. AB - BACKGROUND: In the 1940s and 1950s retinopathy of prematurity (ROP) was the single commonest cause of blindness in children in many industrialised countries; it now accounts for only 6-18% of blindness registrations. It is not known what proportion of blindness is due to ROP in countries that do not have blindness registers. Information on blindness in children in these countries can be obtained by examining children in schools for the blind. METHODS: Between 1991 and 1996, 4121 children in 23 countries with a visual acuity in the better eye of less than 6/60 were examined with a standard method. FINDINGS: The proportion of severe visual impairment or blindness due to ROP ranged from 0% in most African countries to 38.6% in Cuba. INTERPRETATION: These data suggest that ROP is becoming a major cause of potentially preventable blindness among children in middle-income countries that have introduced neonatal intensive-care services for preterm and low-birthweight babies. PMID- 9217714 TI - Thromboxane antagonism and cough induced by angiotensin-converting-enzyme inhibitor. AB - BACKGROUND: The increased prostaglandin synthesis that might follow stimulation of the arachidonic acid cascade by angiotensin-converting-enzyme inhibition (ACE I) has been suggested to underlie the appearance of cough on ACE-I treatment. We investigated whether the prostanoid thromboxane was involved. METHODS: Nine patients with essential hypertension who had cough after enalapril 20 mg once a day (coughers) were treated, while continuing the enalapril, in a double-blind crossover study with placebo or picotamide, 600 mg twice daily. Picotamide is a platelet antiaggregant that acts through both inhibition of thromboxane synthase and thromboxane-receptor antagonism. Thirteen hypertensive patients with no history of ACE-I-induced cough were also treated with enalapril and served as controls. Cough frequency was measured by a visual analogue scale and by a daily cough diary. 24 h urinary recovery of 11-dehydro-thromboxane-B2 and 6-keto-PGF1 alpha were measured to assess any changes in endoperoxide metabolism during the study periods. FINDINGS: 11-dehydro-thromboxane-B2 (TXB2) recovery was significantly reduced by picotamide, which led to the disappearance of cough in eight patients within 72 h. Picotamide urinary recovery data suggested incomplete absorption in the non-responder. At baseline and after rechallenge with enalapril, 11-dehydro-TXB2 excretion was in the same range in the controls and in the coughers, but the latter showed significantly lower excretion of 6-keto-PGF1 alpha, and their ratio of 11-dehydroTXB2 to 6-keto-PGF1 alpha was twice that of the controls (1.40 [95% CI 0.86-1.95] vs 0.61 [0.37-0.84]). INTERPRETATION: A thromboxane antagonist is effective in ACE-I-induced cough. An imbalance between thromboxane and prostacyclin may represent a marker of patients susceptible to ACE-I-induced cough. PMID- 9217716 TI - Exposure of infants to phyto-oestrogens from soy-based infant formula. AB - BACKGROUND: The isoflavones genistein, daidzein, and their glycosides, found in high concentrations in soybeans and soy-protein foods, may have beneficial effects in the prevention or treatment of many hormone-dependent diseases. Because these bioactive phyto-oestrogens possess a wide range of hormonal and non hormonal activities, it has been suggested that adverse effects may occur in infants fed soy-based formulas. METHODS: To evaluate the extent of infant exposure to phyto-oestrogens from soy formula, the isoflavone composition of 25 randomly selected samples from five major brands of commercially available soy based infant formulas were analysed, and the plasma concentrations of genistein and daidzein, and the intestinally derived metabolite, equol, were compared in 4 month-old infants fed exclusively soy-based infant formula (n = 7), cow-milk formula (n = 7), or human breast-milk (n = 7). FINDINGS: All of the soy formulas contained mainly glycosides of genistein and daidzein, and the total isoflavone content was similar among the five formulas analysed and was related to the proportion of soy isolate used in their manufacture. From the concentrations of isoflavones in these formulas (means 32-47 micrograms/mL), the typical daily volume of milk consumed, and average bodyweight, a 4-month-old infant fed soy formula would be exposed to 28-47 per day, or about 4.5-8.0 mg/kg bodyweight per day, of total isoflavones. Mean (SD) plasma concentrations of genistein and daidzein in the seven infants fed soy-based formulas were 684 (443) ng/mL and 295 (60) ng/mL, respectively, which was significantly greater (p < 0.05) than in the infants fed either cow-milk formulas (3.2 [0.7] and 2.1 [0.3] ng/mL), or human breast-milk (2.8 [0.7] and 1.4 [0.1] ng/mL), and an order of magnitude higher per bodyweight than typical plasma concentrations of adults consuming soy foods. INTERPRETATION: The daily exposure of infants to isoflavones in soy infant formulas is 6-11 fold higher on a bodyweight basis than the dose that has hormonal effects in adults consuming soy foods. Circulating concentrations of isoflavones in the seven infants fed soy-based formula were 13000-22000 times higher than plasma oestradiol concentrations in early life, and may be sufficient to exert biological effects, whereas the contribution of isoflavones from breast milk and cow-milk is negligible. PMID- 9217717 TI - A mechanic who coughed up blood for 15 months. PMID- 9217715 TI - Serious reactions after mass treatment of onchocerciasis with ivermectin in an area endemic for Loa loa infection. AB - BACKGROUND: In 1995, the World Bank launched an African Programme for Onchocerciasis Control to eliminate Onchocerca volvulus disease from 19 African countries by means of community-based ivermectin treatment (CBIT). Several cases of encephalopathy have been reported after ivermectin in people heavily infected with microfilariae of Loa loa (loiasis). We assessed the incidence of serious events in an area where onchocerciasis and loiasis are both endemic. METHODS: Ivermectin (at 150 micrograms/kg) was given to 17877 people living in the Lekie area of Cameroon. 50 microL samples of capillary blood were taken during the daytime before treatment from all adults (aged > or = 15 years), and the numbers of L loa and Mansonella perstans microfilariae in them were counted. Patients were monitored for 7 days after treatment. Adverse reactions were classified as mild, marked, or serious. Serious reactions were defined as those associated with a functional impairment that required at least a week of full-time assistance to undertake normal activities. We calculated the relative risk of developing marked or serious reactions for increasing L loa microfilarial loads. Risk factors for serious reactions were identified and assessed with a logistic regression model. FINDINGS: 20 patients (0-11%) developed serious reactions without neurological signs but associated with a functional impairment lasting more than a week. Two other patients were in coma for 2-3 days, associated with L loa microfilariae in cerebrospinal fluid. Occurrence of serious reactions was related to the intensity of pretreatment L loa microfilaraemia. The relative risk of developing marked or serious reactions was significantly higher when the L loa load exceeded 8000 microfilariae/mL; for serious reactions, the risk is very high (odds ratio > 1000) for loads above 50000 microfilariae/mL. INTERPRETATION: Epidemiological surveys aimed at assessing the intensity of infection with L loa microfilariae should be done before ivermectin is distributed for onchocerciasis control in areas where loiasis is endemic. In communities at risk, monitoring procedures should be established and adhered to during CBIT so that people developing serious reactions may receive appropriate treatment. PMID- 9217718 TI - Percutaneous MRI-guided laser-induced thermotherapy for hepatic metastases for colorectal cancer. PMID- 9217719 TI - Is the cholesterol-lowering effect of simvastatin influenced by CYP2D6 polymorphism? PMID- 9217720 TI - Prevention of neural-tube defects. PMID- 9217721 TI - Does the use of chopsticks for eating transmit Helicobacter pylori? PMID- 9217722 TI - Acute non-cardiogenic lung injury in benign tertian malaria. PMID- 9217723 TI - Extracorporeal photopheresis and interferon-alpha in advanced cutaneous T-cell lymphoma. PMID- 9217724 TI - An experimental model of necrotising enterocolitis. PMID- 9217725 TI - Hair abnormalities in Alzheimer's disease. PMID- 9217726 TI - Polymyalgia rheumatica. PMID- 9217727 TI - Country profile: United Kingdom. PMID- 9217728 TI - Hong Kong, 1894: the role of James A Lowson in the controversial discovery of the plague bacillus. PMID- 9217729 TI - Landmines: time for a ban. PMID- 9217730 TI - Medical ethics: the Israeli Medical Association. PMID- 9217731 TI - Rwanda: physician complicity and rebuilding the medical community. PMID- 9217732 TI - Should paediatric intensive care be centralised? Steering Group for Pan Thames Study of Critically ill Children. PMID- 9217733 TI - Should paediatric intensive care be centralised. PMID- 9217734 TI - Should paediatric intensive care be centralised. PMID- 9217736 TI - Should paediatric intensive care be centralised. PMID- 9217735 TI - Should paediatric intensive care be centralised. PMID- 9217737 TI - Fieldwork in the tropics: duplicate frozen samples. PMID- 9217738 TI - Are Hib booster vaccinations redundant? PMID- 9217739 TI - Diagnosis of new variant Creutzfeldt-Jakob disease by tonsil biopsy. PMID- 9217740 TI - Dangers of non-sedating antihistamines. PMID- 9217741 TI - Dangers of non-sedating antihistamines. PMID- 9217742 TI - The only good Helicobacter pylori is a dead Helicobacter pylori. PMID- 9217743 TI - The only good Helicobacter pylori is a dead Helicobacter pylori. PMID- 9217745 TI - Disclosing conflicts of interest. PMID- 9217744 TI - The only good Helicobacter pylori is a dead Helicobacter pylori. PMID- 9217746 TI - Passive smoking. PMID- 9217747 TI - European research on Alzheimer's disease. PMID- 9217748 TI - Zolpidem in Parkinson's disease. PMID- 9217749 TI - Injury to diabetic feet by thumb tacks. PMID- 9217750 TI - In the shadow of epilepsy. PMID- 9217751 TI - ACE-inhibitors: panacea for progressive renal disease. PMID- 9217752 TI - Colon adenomatous polyps--do they grow inward? PMID- 9217753 TI - Whither digitalis? PMID- 9217754 TI - Microbubble echo-enhancers: a new direction for ultrasound? PMID- 9217755 TI - Could diuretics in hypoxic states be bad for the pulmonary circulation? PMID- 9217756 TI - Randomised placebo-controlled trial of effect of ramipril on decline in glomerular filtration rate and risk of terminal renal failure in proteinuric, non diabetic nephropathy. The GISEN Group (Gruppo Italiano di Studi Epidemiologici in Nefrologia) AB - BACKGROUND: In diabetic nephropathy, angiotensin-converting-enzyme (ACE) inhibitors have a greater effect than other antihypertensive drugs on proteinuria and the progressive decline in glomerular filtration rate (GFR). Whether this difference applies to progression of nondiabetic proteinuric nephropathies is not clear. The Ramipril Efficacy in Nephropathy study of chronic nondiabetic nephropathies aimed to address whether glomerular protein traffic influences renal-disease progression, and whether an ACE inhibitor was superior to conventional treatment, with the same blood-pressure control, in reducing proteinuria, limiting GFR decline, and preventing endstage renal disease. METHODS: In this prospective double-blind trial, 352 patients were classified according to baseline proteinuria (stratum 1: 1-3 g/24 h; stratum 2: > or = 3 g/24 h), and randomly assigned ramipril or placebo plus conventional antihypertensive therapy targeted at achieving diastolic blood pressure under 90 mm Hg. The primary endpoint was the rate of GFR decline. Analysis was by intention to treat. FINDINGS: At the second planned interim analysis, the difference in decline in GFR between the ramipril and placebo groups in stratum 2 was highly significant (p = 0.001). The Independent Adjudicating Panel therefore decided to open the randomisation code and do the final analysis in this stratum (stratum 1 continued in the trial). Data (at least three GFR measurements including baseline) were available for 56 ramipril-assigned patients and 61 placebo-assigned patients. The decline in GFR per month was significantly lower in the ramipril group than the placebo group (0.53 [0.08] vs 0.88 [0.13] mL/min, p = 0.03). Among the ramipril-assigned patients, percentage reduction in proteinuria was inversely correlated with decline in GFR (p = 0.035) and predicted the reduction in risk of doubling of baseline creatinine or endstage renal failure (18 ramipril vs 40 placebo, p = 0.04). The risk of progression was still significantly reduced after adjustment for changes in systolic (p = 0.04) and diastolic (p = 0.04) blood pressure, but not after adjustment for changes in proteinuria. Blood-pressure control and the overall number of cardiovascular events were similar in the two treatment groups. INTERPRETATION: In chronic nephropathies with proteinuria of 3 g or more per 24 h, ramipril safely reduces proteinuria and the rate of GFR decline to an extent that seems to exceed the reduction expected for the degree of blood-pressure lowering. PMID- 9217757 TI - Sentinel-node biopsy to avoid axillary dissection in breast cancer with clinically negative lymph-nodes. AB - BACKGROUND: Axillary lymph-node dissection is an important staging procedure in the surgical treatment of breast cancer. However, early diagnosis has led to increasing numbers of dissections in which axillary nodes are free of disease. This raises questions about the need for the procedure. We carried out a study to assess, first, whether a single axillary lymph node (sentinel node) initially receives malignant cells from a breast carcinoma and, second, whether a clear sentinel node reliably forecasts a disease-free axilla. METHODS: In a consecutive series of 163 women with operable breast carcinoma, we injected microcolloidal particles of human serum albumin labelled with technetium-99m. This tracer was injected subdermally, close to the tumour site, on the day before surgery, and scintigraphic images of the axilla and breast were taken 10 min, 30 min, and 3 h later. A mark was placed on the skin over the site of the radioactive node (sentinel node). During breast surgery, a hand-held gamma-ray detector probe was used to locate the sentinel node, and make possible its separate removal via a small axillary incision. Complete axillary lymphadenectomy was then done. The sentinel node was tagged separately from other nodes. Permanent sections of all removed nodes were prepared for pathological examination. FINDINGS: From the sentinel node, we could accurately predict axillary lymph-node status in 156 (97.5%) of the 160 patients in whom a sentinel node was identified, and in all cases (45 patients) with tumours less than 1.5 cm in diameter. In 32 (38%) of the 85 cases with metastatic axillary nodes, the only positive node was the sentinel node. INTERPRETATION: In the large majority of patients with breast cancer, lymphoscintigraphy and gamma-probe-guided surgery can be used to locate the sentinel node in the axilla, and thereby provide important information about the status of axillary nodes. Patients without clinical involvement of the axilla should undergo sentinel-node biopsy routinely, and may be spared complete axillary dissection when the sentinel node is disease-free. PMID- 9217758 TI - Reduction of concentration of HIV-1 in semen after treatment of urethritis: implications for prevention of sexual transmission of HIV-1. AIDSCAP Malawi Research Group. AB - BACKGROUND: Transmission of HIV-1 is predominantly by heterosexual contact in sub Saharan Africa, where sexually transmitted diseases (STDs) are also common. Epidemiological studies suggest that STDs facilitate transmission of HIV-1, but the biological mechanism remains unclear. We investigated the hypothesis that STDs increase the likelihood of transmission of HIV-1 through increased concentration of the virus in semen. METHODS: HIV-1 RNA concentrations were measured in seminal and blood plasma from 135 HIV-1-seropositive men in Malawi; 86 had urethritis and 49 controls did not have urethritis. Men with urethritis received antibiotic treatment according to the guidelines of the Malawian STD Advisory Committee. Samples were analysed at baseline and at week 1 and week 2 after antibiotic therapy in urethritis patients, and at baseline and week 2 in the control group. FINDINGS: HIV-1-seropositive men with urethritis had HIV-1 RNA concentrations in seminal plasma eight times higher than those in seropositive men without urethritis (12.4 vs 1.51 x 10(4) copies/mL, p = 0.035), despite similar CD4 counts and concentrations of blood plasma viral RNA. Gonorrhoea was associated with the greatest concentration of HIV-1 in semen (15.8 x 10(4) copies/mL). After the urethritis patients received antimicrobial therapy directed against STDs, the concentration of HIV-1 RNA in semen decreased significantly (from 12.4 x 10(4) copies/mL to 8.91 x 10(4) copies/mL at 1 week [p = 0.03] and 4.12 x 10(4) copies/mL at 2 weeks [p = 0.0001]). Blood plasma viral RNA concentrations did not change. There was no significant change in seminal plasma HIV-1 RNA concentrations during the 2-week period in the control group (p = 0.421). INTERPRETATION: These results suggest that urethritis increases the infectiousness of men with HIV-1 infection. HIV-1-control programmes, which include detection and treatment of STDs in patients already infected with HIV-1, may help to curb the epidemic. Targeting of gonococcal urethritis may be a particularly effective strategy. PMID- 9217759 TI - Safety of surfaces and equipment for children in playgrounds. AB - BACKGROUND: The safety of playgrounds is important to protect children from injury, but studies are mostly done mainly under laboratory conditions without epidemiological data. We investigated the safety of different playground surfaces, and types and heights of equipment in public playgrounds in the City of Cardiff, UK. METHODS: We did a correlational study of 330 children aged between 0 and 14 years. All children were hurt when playing in playgrounds in Cardiff and presented to the Accident and Emergency Department in Cardiff Royal Infirmary during summer (April to September) 1992 and 1993, and the whole of 1994. We studied the children's hospital records to establish the type of injury and interviewed their parents to find out the playground and type of equipment involved. The main outcome measures were the number of children injured whilst playing, and injury rates per observed number of children on different surfaces, types, and heights of equipment. FINDINGS: Children sustained significantly more injuries in playgrounds with concrete surfaces than in those with bark or rubberised surfaces (p < 0.001). Playgrounds with rubber surfaces had the lowest rate of injury, with a risk half that of bark and a fifth of that of concrete. Bark surfaces were not significantly more protective against arm fractures than concrete. Most injuries were equipment related. Injury risk due to falls from monkey bars (suspended parallel bars or rings between which children swing) was twice that for climbing-frames and seven times that for swings or slides. The height of the equipment correlated significantly with the number of fractures (p = 0.005) from falls. INTERPRETATION: Rubber or bark surfacing is associated with a low rate of injuries and we support their use in all public playgrounds. Bark alone is insufficient, however, to prevent all injuries, particularly arm fractures. Rubberised impact-absorbing surfaces are safer than bark. We believe that playing on monkeys bars increases the risk of injury in playgrounds and that they should generally not be installed. Safety standards should be based on physical and epidemiological data. Our data suggest that the proposed raising of the maximum fall height from 2.5 m to 3.0 m in Europe is worrying. PMID- 9217761 TI - A 44-year-old woman with diarrhoea and double vision. PMID- 9217760 TI - Distinction of idiopathic Parkinson's disease from multiple-system atrophy by stimulation of growth-hormone release with clonidine. AB - BACKGROUND: Idiopathic Parkinson's disease is a common neurodegenerative disease that is difficult to distinguish from other parkinsonian syndromes such as multiple-system atrophy (MSA). In MSA, autonomic dysfunction is common and is associated with either parkinsonian or cerebellar features, or both. Differentiation of idiopathic Parkinson's disease from MSA is important because prognosis, complications, and response to therapy vary according to disorder. Our aim was to find out whether clonidine/growth hormone (GH) testing distinguishes idiopathic Parkinson's disease from MSA. METHODS: Clonidine is a centrally active alpha 2-adrenoceptor agonist that raises concentrations of GH in serum in healthy people and those with pure autonomic failure (with peripheral lesions), but not in those with MSA (with a central autonomic deficit). We investigated the effects of clonidine on 14 people with idiopathic Parkinson's disease (without autonomic deficits). 31 people with MSA of the three different clinical forms (parkinsonian, cerebellar, and mixed), 19 people with pure autonomic failure, and 27 healthy participants. In nine people with parkinsonian MSA (MSA-P), the GH response to levodopa was also assessed. FINDINGS: Clonidine raised serum GH concentrations in patients with idiopathic Parkinson's disease (median increase 8.98 [IQR 6.6-16.6] mU/L), normal participants (13.2 [7.0-18.6] mU/L), and patients with pure autonomic failure (12.5 [5.6-18.2] mU/L). In those with MSA who had central autonomic failure, GH concentrations were unchanged (MSA-P; 0.41 [-0.30 to 2.09] mU/L and cerebellar MSA [MSA-C] 1.67 [0-4.49] mU/L). The GH response to clonidine in idiopathic Parkinson's disease was significantly different from that in MSA-P (p < 0.0002). In MSA-P, the dopamine precursor levodopa raised GH concentrations (from mean 2.7 [SE 1.0] mU/L to mean 18.2 [6.0] mU/L, p < 0.05) and GH-releasing hormone (GHRH) concentrations (from mean 20.6 [3.25] ng/L to mean 68.0 [10.6] ng/L, p < 0.05), excluding dysfunction of pituitary somatotrophs or GHRH neurons as a cause for the absent GH response to clonidine in MSA. INTERPRETATION: The GH responses to clonidine clearly differentiated idiopathic Parkinson's disease from MSA-C and MSA-P. Together with the levodopa studies they indicated a specific alpha 2-adrenoceptor-hypothalamic deficit in MSA. The clonidine-GH test may provide further insight into central neurotransmitter and alpha 2-adrenoceptor-hypothalamic abnormalities in MSA. PMID- 9217762 TI - Randomised trial of temporary cardiac pacing with semirigid and balloon-flotation electrode catheters. PMID- 9217763 TI - Dual effect of CCR5 delta 32 gene deletion in HIV-1-infected patients. Copenhagen AIDS Study Group. PMID- 9217764 TI - Late-onset fatal acute leucoencephalopathy in liver transplant recipient. PMID- 9217765 TI - Multidrug resistance protein in recurrent breast cancer. PMID- 9217766 TI - Complement C3 and factor B cerebrospinal fluid concentrations in bacterial and aseptic meningitis. PMID- 9217767 TI - Nonsense mutation of prostacyclin synthase gene in a family. PMID- 9217768 TI - Colonic perforation and serosal tears associated with colonoscopy. PMID- 9217769 TI - USA outlines standard of care for HIV/AIDS. PMID- 9217770 TI - The alcohol withdrawal syndrome. PMID- 9217771 TI - Methicillin-resistant Staphylococcus aureus and vancomycin-resistant enterococci: therapeutic realities and possibilities. AB - During the past decade much effort has been devoted worldwide to limiting the spread of methicillin-resistant Staphylococcus aureus. However, the recent emergence of almost untreatable vancomycin-resistant enterococci has led to a new and unexpected public health problem in hospitals and the community. Moreover, the threat of transfer of glycopeptide resistance to S aureus means that development of alternative antimicrobial strategies has become urgent. Whereas major advances have been made in our understanding of methicillin and vancomycin resistance mechanisms, we still need to identify the sources and reservoirs of the genetic determinants of resistance and to discover how they disseminate in the environment. The outcome of the battle between antimicrobials and bacteria is still uncertain, but the challenge is worth meeting. PMID- 9217772 TI - Euthanasia and the potential adverse effects for Northern Territory aborigines. PMID- 9217773 TI - Invasive cervical cancer after treatment for cervical intraepithelial neoplasia. PMID- 9217774 TI - Invasive cervical cancer after treatment for cervical intraepithelial neoplasia. PMID- 9217775 TI - Invasive cervical cancer after treatment for cervical intraepithelial neoplasia. PMID- 9217776 TI - Pneumococcal bacteraemia in Sweden. PMID- 9217777 TI - Bayesian interim analysis of randomised trials. PMID- 9217778 TI - Bayesian interim analysis of randomised trials. PMID- 9217779 TI - Bayesian interim analysis of randomised trials. PMID- 9217780 TI - Genetic influence on cytokine production in meningococcal disease. PMID- 9217781 TI - Sick-building syndrome. PMID- 9217782 TI - Sick-building syndrome. PMID- 9217783 TI - Replication of zolpidem test for catatonia in an adolescent. PMID- 9217784 TI - Antiplatelet therapy in ischaemic events. PMID- 9217785 TI - AIDS epidemic in Kaliningrad. PMID- 9217786 TI - Health inequality: the UK's biggest health issue. PMID- 9217787 TI - Misdiagnosis of multiple sclerosis and beta-interferon. PMID- 9217789 TI - Quality control of cochlear implants. PMID- 9217788 TI - Health care under the Taliban. PMID- 9217790 TI - Effects of pentachlorophenol exposure. PMID- 9217791 TI - Future directions in treatment of amblyopia. PMID- 9217792 TI - Are we overinvestigating appendicitis. PMID- 9217793 TI - Escherichia coli O157 in abattoirs. PMID- 9217794 TI - How to "peer review" a medical journal manuscript. AB - BACKGROUND: The accuracy, validity, and appropriateness of what gets published in the medical literature depends upon a pool of competent peer referees. However, the requisite skills needed to become such a peer are not taught at the residency training program level or subsequently by the journals themselves, who offer no training and little written instruction. OBJECTIVE: To outline a logical, orderly approach to the task of reviewing a "raw" medical manuscript. By breaking the overall endeavor down into smaller, step-by-step components, the novice reviewer should attain the direction and skills to complete a review with confidence. METHODS: Explore the role of the reviewer, discuss the philosophy of review, identify the key elements to look for when reading and rereading the manuscript, and outline a orderly approach to the decision making process. CONCLUSIONS: A "peer" is a physician with expertise on the subject under scrutiny who spends sufficient time and thought to fulfill two main obligations: to render an honest, unbiased decision on whether or not the manuscript should be published and, if it is acceptable, to help make it better. It is possible to perform the review process in a relatively simple, organized, and logical manner. PMID- 9217795 TI - Scar revision. AB - BACKGROUND: Scars of cosmetic or functional importance may form following cutaneous surgery, trauma, or inflammation. Many factors interplay in the formation of these scars. Knowledge and proper planning can help eliminate these consequences. Various scar revision techniques, both surgical and nonsurgical, are now available for treating undesirable scarring. OBJECTIVE: To review the various scar revision options for the various types of scars. LEARNING OBJECTIVE: After reading this review the participant should have a better approach and understanding of the appropriate scar revision techniques. PMID- 9217796 TI - The molecular basis of xeroderma pigmentosum. AB - BACKGROUND: Xeroderma pigmentosum is an extremely rare, autosomal recessive disease characterized by a more than 1000-fold increase in nonmelanoma skin cancer. Individuals with this disease can be divided into eight complementation groups: A-G and V for variant. Each one represents a different genetic defect in DNA repair. OBJECTIVE: To review the molecular basis of xeroderma pigmentosum. RESULTS: Deficiencies in various gene products in the nucleotide excision repair pathway cause xeroderma pigmentosum in complementation groups A-G. The molecular basis of the variant group remains to be elucidated. CONCLUSIONS: Research into the genetic defects underlying xeroderma pigmentosum have led to an increased understanding of nucleotide excision repair. PMID- 9217797 TI - Lip augmentation with preserved fascia lata. AB - BACKGROUND: Presently available techniques for lip augmentation have an assortment of limitations. OBJECTIVE: To provide a safe, reliable method of lip augmentation on a long-term basis. METHODS: Through a stab incision in each quadrant, chips of human cadaver, banked fascia lata were inserted into intralabial pockets. RESULTS: Fascia lata grafting proved to be a simple effective technique of lip enhancement. Over the period of follow-up, enhancement was evident in most cases and no allergic reactions or infections occurred. Lip motion was satisfactory and paresthesia were minor. CONCLUSIONS: Fascia lata grafting is a simple, controlled technique for graded lip enhancement. PMID- 9217798 TI - The use of antidesmoglein stains in Mohs micrographic surgery. A potential aid for the differentiation of basal cell carcinoma from horizontal sections of the hair follicle and folliculocentric basaloid proliferation. AB - BACKGROUND: Histopathologic differentiation between benign and malignant tissue is of utmost importance for the Mohs surgeon. Folliculocentric basaloid proliferation (FBP) shares many histologic features with basal cell carcinoma (BCC). It is most commonly associated with tumors of areas with abundant hair follicles such as nasal and perinasal skin. Residual BCC incorrectly identified as a horizontally sectioned hair follicle undoubtedly increases the risk of tumor recurrence. Excision of additional layers of normal tissue to remove "funny looking follicles" may have profound impacts on tissue conservation, preservation of function, and cosmesis. Electron microscope studies of BCC revealed a significant reduction of desmosomes compared with normal basal cells and hair follicle keratinocytes. OBJECTIVE: This study has assessed the potential of rapid staining with monoclonal antidesmoglein antibody (33-3D) to discriminate between BCC, horizontally sectioned hair follicles, and FBP. METHODS: A rapid immunoperoxidase technique with 33-3D antidesmoglein antibody was performed on Mohs frozen sections. We selected 18 patients with BCC of nasal and perinasal locations where histologic discrimination between residual tumor and tumor-free margins with FBP or horizontally sectioned hair follicle was equivocal. RESULTS: Fourteen sections disclosed the preservation of desmoglein marker delineating the cell membranes ("perimembranous" pattern) consistent with normal hair follicles. The sections were identified as tumor-free and no additional stages were performed. The remaining four sections revealed absent perimembranous pattern but presence of diffuse cytoplasmic staining. These were diagnosed as positive for residual BCC requiring the excision of another layer of tissue to obtain tumor free margins. A follow-up period ranging from 6 to 24 months revealed no instance of recurrent disease. CONCLUSION: Rapid detection of desmoglein with 33-3D antibody is a promising tool for discrimination between residual BCC and FBP or horizontally sectioned hair follicles. It may enhance the sensitivity of Mohs surgery by disclosing the hidden foci of BCC, thus preventing tumor recurrence and unnecessary excision of normal tissue. PMID- 9217799 TI - Experience with a make-it-yourself scalp extender with hooks. AB - BACKGROUND: Silicone scalp extenders have been shown to facilitate the process of scalp reduction surgery. OBJECTIVE: This report demonstrates how scalp extender devices can easily be made in the physician's office out of materials that are commercially available. METHODS: A scalp extender is made by bonding a narrow strip of Dacron-reinforced Duralastic silicone sheeting to each end of a long strip of plain Duralastic silicone sheeting. A stainless steel plate of hooks is then secured to each end of the extender. At the time of the first standard scalp reduction surgery, one extender device about 3.0 x 5.0 cm is placed in the subgaleal plane, in the transauricular direction, with hooks piercing the galea 1 cm caudad to the dense-in-quantity hair fringe margin. The circumferencial approach will accommodate the placement of a second extender device in the anterior-posterior direction to elevate the posterior hair fringe. To prevent adhesion of the galea to the periosteum around the devices, a sheet of plain silicone 15 x 20 cm is placed over the periosteum before the extenders are inserted. The extenders are replaced with shorter devices 3-4 weeks later. RESULTS: Usually the average width of bald scalp can be closed fringe to fringe in three operations as compared with five or six operations when using standard scalp reduction procedures. CONCLUSIONS: The scalp extender device described can be custom made in many different sizes. Its use has significantly changed the author's surgical treatment of pattern alopecia. PMID- 9217800 TI - Fractional cryosurgery. A new technique for basal cell carcinoma of the eyelids and periorbital area. AB - BACKGROUND: Cryosurgery is an established method to treat malignant tumors of the eyelids and periorbital area. Nevertheless, it has been abandoned for tumors greater than 10 mm, because it gives irregular esthetic results and, in some cases, lagophthalmos. OBJECTIVE: To devise a new method for the treatment of such tumors. METHOD: Fractional cryosurgery is performed in stages: the center of the lesion is frozen, resulting in a reduction of the tumor; this procedure is repeated, as necessary, until the lesion's diameter is smaller than 10 mm; the standard cryosurgical procedure is then carried out. RESULTS: The treatment of the first 20 basal cell carcinomas with diameters between 10 and 24 mm is described, with excellent clinical and cosmetic results. CONCLUSION: With fractional cryosurgery, the final scar bears no relation to the size of the original tumor but, instead, corresponds to the size of the lesion preceding the final cryosurgical procedure. PMID- 9217801 TI - Cryosurgical treatment of professional chronic radiodermatitis. AB - BACKGROUND: Chronic x-ray dermatitis in professionals is a frequent problem for doctors in our country due to the fact that many of them widely used radiotherapy without any protection 15-20 years ago. Surgery has been the most accepted treatment, though it generally decreases hand function. OBJECTIVE: Up to now, cryosurgery was not usually considered as a possible treatment if the lesions were located on fingers. In this study, the advantages of cryosurgery for the treatment of professional chronic radiodermatitis with incipient pretumoral lesions are emphasized. METHODS: Cryosurgery was performed on six patients affected with chronic professional radiodermatitis that showed keratomas and ulcerations, using both spray (keratomas) and a probe 0.5 cm in diameter (ulcerations, in situ squamous cell carcinoma). Nerve block anesthesia with mepivacaine 1% was used in all cases. Before the treatment, all suspected lesions were biopsied; if invasive squamous cell carcinoma was revealed in the dermatopathological study, the patient was rejected. Variables such as blister and necrosis formation, pain, and achromatic, sensibility, and mobility disorders were studied. The follow-up period was 2 years. RESULTS: Immediate postoperative results showed great pain and blistering in all cases. Residual achromias were observed early postoperatively in all cases, but were repigmented 1 year after therapy in four cases (66%). Sensory alterations (hypo- and hyperthesias) were found in four cases (66%) 1 month after treatment, although this complication was not observed 6 months after treatment. Finger mobility was perfect in all cases 2 months after treatment, and there was no recurrence in any case after 2 years of follow-up. CONCLUSIONS: We believe cryosurgery must be considered as an excellent treatment for professional chronic radiodermatitis with keratomas, ulcerations, and incipient squamous cell carcinomas. Its use may prevent further dramatic surgical treatment, like amputations, allowing the preservation of finger function. PMID- 9217802 TI - Squamous cell carcinoma in situ arising within clear cell acanthoma. AB - BACKGROUND: A brief discussion of the subject. OBJECTIVE: The purpose of the work to be described. METHODS: How the work was performed. RESULTS: The outcome of the work. CONCLUSION: The conclusion that can be reached based on the work described. PMID- 9217803 TI - Carcinoma of the mammary crease simulating rodent ulcer basal cell carcinoma. Report of a case with immunohistochemical analysis. PMID- 9217804 TI - Diagnostic problems of desmoplastic melanoma in a boy with xeroderma pigmentosa. PMID- 9217805 TI - How many skin cancers require Mohs micrographic surgery? PMID- 9217806 TI - Problems of tumescent anesthesia for dermabrasion. PMID- 9217807 TI - Hebiclens keratitis and the TCA Masque. PMID- 9217808 TI - What of the clinical significance of the cryoimmune response? PMID- 9217809 TI - In vitro reconstructed skin models for wound coverage in deep burns. PMID- 9217810 TI - Immunohistochemical analysis of the skin in junctional epidermolysis bullosa using laminin 5 chain specific antibodies is of limited value in predicting the underlying gene mutation. AB - The anchoring filament protein laminin 5 is composed of three polypeptide chains (alpha 3, beta 3 and gamma 2) each encoded by separate genes (LAMA3, LAMB3 and LAMC2, respectively). Mutations in any of these three genes may give rise to the autosomal recessive blistering skin disease, junctional epidermolysis bullosa. At present, there is no easy way of predicting which of these three genes might harbour the pathogenetic laminin 5 mutations in a case of junctional epidermolysis bullosa. In this study, we assessed whether immunohistochemistry might be helpful in this regard. We performed immunohistochemical labelling of the dermal-epidermal junction using alpha 3, beta 3 and gamma 2 chain-specific antibodies in 11 patients with junctional epidermolysis bullosa, in whom the laminin 5 mutations had been previously delineated. Although, labelling for the laminin 5 chain bearing the mutations was attenuated or undetectable in all cases, a complete absence of labelling or a reduction in the staining intensity for the other two chains was also seen in all cases. The results showed that immunohistochemical labelling of the dermal-epidermal junction using alpha 3, beta 3 and gamma 2 chain-specific antibodies is not a specific indicator for which of the laminin 5 chain genes contains the pathogenetic mutations, and is therefore unreliable in screening for individual laminin 5 gene mutations in cases of junctional epidermolysis bullosa. PMID- 9217811 TI - Detection of Epstein-Barr virus in primary cutaneous amyloidosis. AB - To determine the association of Epstein-Barr virus (EBV) with primary cutaneous amyloidosis (PCA), a retrospective study was conducted on skin tissue from 27 Chinese patients with lichen amyloidosus and macular amyloidosis. In situ hybridization with oligonucleotide probes was used to detect the expression of EBV-encoded RNAs (EBERs). Eleven of 27 cases (40.7%) were found to contain the EBV genome. No EBV genome was detected in the skin of the control groups, including three cases of secondary cutaneous amyloidosis, two cases of primary systemic amyloidosis, and four cases of lichen simplex chronicus. Our study showed no correlation between the presence of EBV in PCA patients and the patients' age, sex, clinical type or severity of the skin lesions. Although our results suggest that EBV may be associated with some cases of PCA, the true aetiological role of EBV in PCA remains unknown. PMID- 9217812 TI - Lack of evidence of human herpesvirus 8 DNA sequences in HIV-negative patients with various lymphoproliferative disorders of the skin. AB - Human herpesvirus 8 (HHV-8) is a new virus which has been reported in Kaposi's sarcoma and some lymphoproliferative disorders such as Castleman's disease and body-cavity-based lymphoma. Because HHV-8 shares homology with Epstein-Barr virus (EBV), we searched for the presence of HHV-8 DNA sequences in various cutaneous T and B-cell lymphoma by the polymerase chain reaction (PCR). Forty-seven HIV negative patients with cutaneous lymphoma or large plaque parapsoriasis were enrolled in the study. For the detection of HHV-8 DNA sequences we used PCR followed by a hybridization with a digoxigenin-labelled probe and nested-PCR. HHV 8 DNA sequences could only be detected in a patient with large plaque parapsoriasis. Our study does not suggest any direct implication of HHV-8 in the pathogenesis of most cutaneous lymphoma. Serological studies will be helpful to appreciate if there is an epidemiological link between HHV-8 and cutaneous lymphomas. PMID- 9217813 TI - Interferon-gamma differentially regulates CD80 (B7-1) and CD86 (B7-2/B70) expression on human Langerhans cells. AB - CD80 (B7-1) and CD86 (B7-2/B70) have recently been identified in cultured human Langerhans cells (LCs), although their role and regulatory properties remain unclear. We present our comparison of the expression of the molecules, mRNAs and the function between CD80 and CD86 in human LCs treated by interferon gamma (IFN gamma). We examined the regulatory properties of CD80 and CD86 expression in human LCs pretreated with IFN-gamma. Flow cytometric analysis indicated that the mean fluorescence intensity of CD86 but not CD80 was enhanced. However, the percentage modulation of both CD80 and CD86 positive cells were significantly up regulated in a dose-dependent manner, after 48-h culturing with IFN-gamma. The regulatory properties of CD80 and CD86 mRNA expressions in human LC were studied using polymerase chain reaction methods. We found that both CD80 and CD86 mRNA of enriched LCs following IFN-gamma pretreatment for 12 h were higher than those without pretreatment. We have demonstrated that the primary allogeneic mixed epidermal cell-lymphocyte reaction induced by human LCs treated by IFN-gamma increased in a dose-dependent manner. There was a 61.5% inhibition by anti-CD86 monoclonal antibody and a 32.5% inhibition by anti-CD80 monoclonal antibody. These data indicate that the CD80 and CD86 expression of human LCs may be differently regulated by IFN-gamma. PMID- 9217814 TI - Functional CD86 (B7-2/B70) is predominantly expressed on Langerhans cells in atopic dermatitis. AB - Recently, we reported the functional expression of CD86 on cultured human Langerhans cells derived from normal epidermis. In the present study, we investigated the expression and function of co-stimulatory molecules in the pathogenesis of atopic dermatitis. In immunohistochemical analysis, CD80 and/or CD86 were detected on dendritic-shaped cells not only in the epidermis but also in the dermis in the inflammatory lesions of atopic dermatitis (n = 12). CD80 was expressed in only five cases (42%), while CD86 was expressed in all cases (100%). These molecules were not detected in normal control subjects (n = 8). In non lesional skin of atopic dermatitis (n = 4), CD86 but not CD80 was detected in one case. CD86 was preferentially induced on dendritic-shaped cells in positive patch test sites to Dermatophagoides pteronyssinus or house dust allergen in atopic dermatitis (n = 4). The CD80- or CD86-positive cells were confirmed as Langerhans cells by double immunostaining using anti-CD1a monoclonal antibody. Neither CD86 nor CD80 was detected on keratinocytes. Similar results of the stronger expression of CD86 over that of CD80 were obtained from psoriasis vulgaris (n = 11) and from contact dermatitis (n = 7), although CD86 was expressed only in 57% of the contact dermatitis cases. The percentage of Langerhans cells positive for CD86 was higher than for CD80, i.e. 48% compared with 9%, respectively, in the epidermis of lesional skin of atopic dermatitis (n = 8). The expression rate of these molecules on Langerhans cells increased in the dermis. To investigate the function of co-stimulatory molecules on Langerhans cells in atopic dermatitis, we conducted an inhibition test with antibodies. Anti-CD86 monoclonal antibody almost completely inhibited T-cell proliferation stimulated with crude extract of D. pteronyssinus in the presence of epidermal cells as antigen-presenting cells, whereas anti-CD80 monoclonal antibody produced less of an inhibitory effect. These data indicate that CD86 expressed on Langerhans cells may play an important part in the pathogenesis of atopic dermatitis. PMID- 9217815 TI - Blockade of costimulatory molecules B7-1 (CD80) and B7-2 (CD86) down-regulates induction of contact sensitivity by haptenated epidermal cells. AB - The hapten, trinitrobenzene sulphonic acid, induced weak B7-1 (CD80) and moderate B7-2 (CD86) expression on Langerhans cells and mRNA expression of both molecules in organ-cultured murine skin. The intradermal injection of hapten-treated epidermal cells induced hapten-specific contact sensitivity in synergic mice. Cells of the keratinocyte cell line, Pam 212, or epidermal cells treated with a mixture of anti-Ia/thy1.2/gamma delta antibody plus complement, did not show any sensitizing ability. When hapten-treated epidermal cells were injected into mice after incubation with anti-B7-2 (CD86) or B7-1 (CD80) antibody the resultant contact sensitivity reaction was decreased to less than 50% of the control reaction, a reduction which was similar to that seen with the anti-ICAM-1 and anti-LFA-1 antibody-induced inhibition of contact sensitivity. Anti-B7-1 (CD80) or anti-B7-2 (CD86) antibody also inhibited hapten-specific lymphocyte proliferation or the allogenic mixed lymphocyte and epidermal cell reaction in vitro, although the inhibitory effect of anti-B7-1 antibody was not as significant as that of anti-B7-2 antibody. These results indicate that costimulatory signals induced by a hapten on epidermal Langerhans cells play an important role in the induction of hapten-specific contact sensitivity in mice. PMID- 9217816 TI - Evidence that activation of protein kinase A inhibits human hair follicle growth and hair fibre production in organ culture and DNA synthesis in human and mouse hair follicle organ culture. AB - We have investigated the possibility that protein kinase A (PKA) may play a part in regulating the activity of human and mouse hair follicles in whole organ culture. Human hair follicles were isolated from facial skin by microdissection, and hair follicle and hair fibre length measurements were made daily during suspension culture. Incubation of human hair follicles with dibutyryl-cAMP (db cAMP) resulted in a dose-dependent inhibition of total cumulative follicle growth (IC50 = 100 mumol/L, 85% inhibition at 1 mmol/L). db-cAMP (0.5 mmol/L) also caused a rapid, partial inhibition of follicular DNA synthesis (20.3% inhibition at 6 h, 48.0% inhibition at 24 h). Human hair follicle growth was inhibited by the phosphodiesterase inhibitors 3-isobutyl-1-methylxanthine and Ro 20-1724, and by the adenylate cyclase activator, forskolin. In addition, db-cAMP inhibited DNA synthesis in organ cultures of whisker follicles isolated from neonatal mice by microdissection. Taken together, these findings indicate that agents which increase cAMP levels are potent inhibitors of human and mouse hair follicle growth, and suggest that PKA may play a part in the regulation of hair follicle activity in vivo. PMID- 9217817 TI - Localization of endothelial proliferation and microvascular expansion in active plaque psoriasis. AB - Expansion of the dermal microvasculature is a prominent feature of psoriasis. Although the pathogenetic process resulting in vascular morphological changes remains unclear, considerable evidence suggests the involvement of angiogenesis. To assess the degree and site of psoriatic microvascular expansion, immunohistochemical studies were performed on paired lesional and non-lesional specimens from 10 patients with active, untreated plaque psoriasis. Five-micron thick sections were labelled with monoclonal antibody JC/70A specific for the endothelial marker CD31, and vascular quantification was achieved using hue saturation-intensity image analysis. Assessment of vasculature in the papillary dermis (superficial plexus) demonstrated a fourfold increase in endothelial surface area of lesional compared with non-lesional skin (P < 0.01), while there was no significant increase in vasculature of the upper reticular dermis. Subsequently, 18-micron thick sections were double-labelled with MIB-1 antibody to the nuclear proliferation marker Ki-67 and JC/70A. Endothelial cell proliferation was identified in the vertical limbs of capillary loops in eight out of 10 lesional biopsies and in no non-lesional biopsies. The endothelial MIB 1 labelling index was 3.1% of total endothelial cells of the superficial plexus. These findings confirm endothelial proliferation underlying psoriatic microvascular expansion, and indicate that this process is limited to a specific site in the dermal capillary bed. PMID- 9217818 TI - Psoriasis patients have highly increased numbers of tryptase-positive mast cells in the duodenal stroma. AB - We have shown that the number of tryptase-positive mast cells in the duodenal mucosa in psoriasis is increased and that a subgroup of psoriasis patients showed elevated levels of antibodies to gliadin (some of whom also had increased lymphocytes in the duodenal epithelium). Duodenal biopsy specimens from 37 patients with psoriasis (eight mild, 13 moderate and 16 severe) and 22 patients with irritable bowel syndrome (IBS) were examined regarding the presence of tryptase + mast cells. Intraepithelial infiltration by lymphocytes was evaluated and scored from 0 to 3. Patients with psoriasis had 131 +/- 58 mast cells/mm2 (mean +/- SD) and those with IBS 28 +/- 18. Only in four of the 37 psoriasis patients was the number within the range of that in the IBS group. There were no signs of stromal inflammation except in one psoriasis patient. No relationship was found between degree of severity of psoriasis and number of mast cells. In 25 of the 37 specimens from psoriasis patients there was no increase in intraepithelial lymphocytes, whereas seven showed a slight increase (score 1-2) and five a moderate increase (score > or = 2-3). The number of tryptase + mast cells was similar in patients with or without increased intraepithelial lymphocytes. The number of mast cells showed no relation to the presence or absence of antibodies to gliadin. We hypothesize that there are at least two types of abnormalities in the duodenal mucosa in psoriasis, one type that is present in most psoriasis patients and characterized by an increase in mast cells and eosinophils, and another that is present in a subgroup of patients with antibodies to gliadin and an increased number of duodenal intraepithelial lymphocytes. The mechanisms underlying the increase in the number of mast cells and its relevance are not yet known. PMID- 9217819 TI - Quantitative analysis of tryptase- and chymase-containing mast cells in atopic dermatitis and nummular eczema. AB - The distribution of mast cells (MCs) containing tryptase (T) and chymase (C) was studied in the non-lesional and lesional skin of 26 patients with atopic dermatitis (AD) and 23 patients with non-atopic nummular eczema (NE), and in the skin of eight healthy controls. T and C activities were demonstrated enzymehistochemically using Z-Gly-Pro-Arg-MNA and Suc-Val-Pro-Phe-MNA as substrates, respectively. The T- and C-containing MCs were counted separately in the epidermis, in contact with the basement membrane, in the papillary dermis and in different dermal levels (0.2 mm each). Also, the C protein was determined immunohistochemically. T-positive MCs were similarly distributed in non-lesional and lesional skin of both AD and NE. The MC number was relatively high in the upper dermis (papillary dermis and levels I and II) of non-lesional and lesional skin of AD. In the upper dermis of non-lesional AD and NE skin and in normal skin, about 50% of T-positive MCs displayed C activity, whereas the percentage in lesional AD and NE skin was only about 30%. In this respect, the non-lesional and lesional samples differed significantly from each other in both dermatoses (in AD p = 0.003; in NE p = 0.002, Students' t-test). In all samples the MC number decreased in the deeper dermal levels, although numerous T-containing MCs were still counted in the deeper dermis (dermal levels IV-VII) of lesional AD and NE skin, differing significantly from the MC number in normal skin (In AD p = 0.005, In NE p = 0.041). In the deeper dermis, the percentage of MCs containing active C was about 70% in non-lesional and lesional AD and NE, and about 90% in normal healthy skin. However, in the upper dermis of non-lesional and lesional skin of both AD and NE, about 80% of all MCs contained the C protein, which differed significantly from the value of 100% in normal skin (p < 0.05). In conclusion, the increased number of T-positive MCs in the upper dermis of non-lesional and lesional AD contributes to promoting inflammation. C apparently loses its activity in the upper dermis of lesional AD and especially in NE. Thus, the enzyme partially lacks its capability to suppress inflammation, such as degradation of neuropeptides and proteins. The dysregulation of these proteinases exists already in non-lesional skin of AD and NE. PMID- 9217820 TI - The location of binding sites of pemphigus vulgaris and pemphigus foliaceus autoantibodies: a post-embedding immunoelectron microscopic study. AB - Pemphigus is a life-threatening autoimmune blistering disease of skin and mucous membranes that has two major subtypes based on clinical and histological features, pemphigus vulgaris (PV) and pemphigus foliaceus (PF). Autoantibodies against the PV antigen (desmoglein 3) and the PF antigen (desmoglein 1) are involved in the pathogenesis of blister formation. In the present study, the location of epitopes recognized by autoantibodies of patients with PV and PF was studied by postembedding immunogold electron microscopy. PV and PF autoantibodies were observed bound predominantly to the intercellular domains of desmosomes, but not to the non-desmosomal keratinocyte cell surface. The relationship between the location of PF antigen and other constitutive desmosomal proteins, desmocollin, desmoplakin and plakoglobin, in normal human skin was investigated using a double immunogold labelling technique. It was observed that PF antigen and desmocollin co-localize within the intercellular domain of the desmosomes. In contrast, the antibodies against desmoplakin and plakoglobin bound predominantly to the intracellular desmosomal attachment plaque with the binding site of the antibody against plakoglobin closer to the desmosomal cell membrane than that of the antibody to desmoplakin. We show that the LR White postembedded immunogold electronmicroscopy technique is convenient and easily applied to studies of autoimmune bullous skin diseases. We have used it to demonstrate the precise localization of the binding sites of PV and PF autoantibodies and their relationship with other constitutive desmosomal proteins. PMID- 9217821 TI - Depletion of stratum corneum intercellular lipid lamellae and barrier function abnormalities after long-term topical corticosteroids. AB - The intercellular lipid lamellae of the stratum corneum (SC) is believed to provide the permeability barrier of the epidermis. Previous functional studies have demonstrated an increase in the transepidermal water loss (TEWL) after long term use of topical corticosteroids (TCS): however, direct morphological confirmation of this barrier abnormality is still lacking. The aim of this study was to determine whether any abnormality could be detected in the structure of the SC intercellular lipid lamellae in patients after long-term TCS. Atrophic skin and untreated normal skin of 10 patients after long-term TCS were examined by transmission electron microscopy using ruthenium tetroxide-fixed tissue for the multilamellar lipid sheets of SC, and oil red O stain for neutral lipids of the SC. Layers of the SC were evaluated by 0.1% methylene blue stain after alkaline expansion, and TEWL was measured by Evaporimeter EP1. The TCS-treated atrophic skin had fewer layers of horny cells, mean 9.4 layers, than the normal control skin, 18 layers (P < 0.001) and increased TEWL of 21.3 g/m2 compared with the control skin TEWL of 6.7 g/m2 (P < 0.01). The mean neutral lipid content of the SC was also significantly lower (P < 0.001). Moreover, ultrastructural studies revealed a marked decrease in both the numbers of intercellular lipid lamellae of SC and membrane-coating granules of stratum granulosum in the atrophic skin. These results suggest that the diminution in the SC intercellular lipid lamellae and SC cell layers play an important part in the pathogenesis of barrier dysfunction after long-term use of TCS. PMID- 9217822 TI - Effect of topical tretinoin on non-sun-exposed human skin connective tissue: induction of tenascin but no major effect on collagen metabolism. AB - The effects of topical tretinoin on collagen synthesis and degradation were studied in 29 volunteers. The subjects applied 0.1% tretinoin cream on their non sun-exposed abdominal skin once a day for 1 week (n = 10) (experiment 1) or twice a day for 2 weeks (n = 8) (experiment 2) or once a day for 2 months (n = 11) (experiment 3). After the treatments, suction blisters were induced and amino terminal propeptides of type I and III procollagens (PINP, PIIINP, respectively) (experiments 1 and 3) and carboxy-terminal propeptide of type I procollagen (PICP) (experiment 2) were assayed as an index of de novo collagen synthesis by radioimmunoassays. Matrix metalloproteases 2 (MMP-2) and 9 (MMP-9) were assayed by the zymography method in experiment 2. In experiment 3, histology, immunohistochemistry of type I and III procollagens, tenascin, mRNA levels of type I collagen alpha 1-chain [alpha 1 (I)], interstitial collagenase (MMP-1), MMP-2, MMP-9 by slot-blot analysis and the levels of alpha 1 (I) collagen mRNA by a quantitative polymerase chain reaction method were studied. The proportional area of elastic fibres visualized in Verhoeff-stained sections was analysed by computerized digital image analysis. The results indicated that treatment with topical tretinoin does not markedly induce de novo synthesis of collagen in vivo or affect matrix metalloproteases. In the immunohistochemical staining, tenascin was increased in the papillary dermis. As it has been suggested that tretinoin could counteract the atrophogenic effect of corticoids on the dermis, the effect of a combination of betamethasone-17-valerate (once a day) and tretinoin (once a day) on the propeptide levels was also studied. Betamethasone alone caused a 60% decrease in the concentrations of PINP and PIIINP, and a similar decrease was found after the combination treatment, indicating that topical tretinoin administered during short treatment periods does not counteract the inhibitory effect of a potent corticoid on collagen propeptides. PMID- 9217823 TI - Treatment of burns and donor sites with human allogeneic keratinocytes grown on acellular pig dermis. AB - The absence of a dermal component predisposes cultured epidermal sheets to instability, contractibility, and makes them difficult to handle. In order to overcome these drawbacks, we developed recombined human/pig skin (RHPS) composed of human keratinocytes cultured on cell-free pig dermis. The original intention to prepare a permanent skin substitute composed of xenodermis and autologous epidermis was not achieved, but it has been proved that RHPS can serve as an effective, ready to use keratinocyte delivery system when applied 'upside-down', i.e. with epidermal cells facing the wound surface. The keratinocyte layer establishes a direct contact with the wound bed, while the dermal layer mechanically protects the wound. Twenty deep dermal burns were grafted with RHPS: 13 (65%) healed completely in 4-14 days, three (15%) healed partially and four (20%) did not heal. Of five full thickness burn wounds only one healed after repeated RHPS grafting within 18 days. Thirty-one (100%) donor sites treated with any of the three forms of RHPS, subconfluent, confluent meshed or confluent unmeshed, healed within 6-8 days compared with 14-18 days in control sites. Seven donor sites (100%) of immunodeficient patients with prolonged wound healing epithelialized in 7-10 days under RHPS compared with 32-90 days in areas treated with tulle gras and dry gauze. PMID- 9217824 TI - Increased level of c-erbB-2/neu/HER-2 protein in cutaneous squamous cell carcinoma. AB - Overexpression of c-erbB-2/neu/HER-2 oncoprotein, a receptor tyrosine kinase, has been demonstrated in a variety of human cancers. To elucidate the involvement of c-erbB-2 in human skin carcinogenesis, we examined expression of the protein in skin samples from five cases of keratoacanthoma (KA), 10 of actinic keratosis (AK), 24 of squamous cell carcinoma (SCC) and 10 of basal cell carcinoma (BCC) and five samples of normal epidermis, using an immunohistochemical method on formalin-fixed, paraffin-embedded sections. Expression of c-erbB-2 was also examined in cultured SCC cell lines, a premalignant cell line and in cultured normal keratinocytes. Normal epidermal cells showed no or very little c-erbB-2 protein, but the covering epidermal layer of some tumours showed a few strongly positive cells. Samples of KA and AK showed barely detectable c-erbB-2 protein in only a few cases. Twenty of the 24 cases of SCC had elevated expression of c-erbB 2 protein, with a tendency to more positive cells in metastatic lesions. Five of the 10 cases of BCC stained for c-erbB-2 but more weakly than those of SCC. Reaction products of the positive cells were seen in the cytoplasm. All three cultured SCC cell lines stained for c-erbB-2 protein more strongly than the premalignant HaCaT or normal keratinocytes. Our results indicate the possible involvement of c-erbB-2 overexpression in the malignant conversion of keratinocytes. PMID- 9217826 TI - Wells' syndrome: a pathogenic role for circulating CD4+CD7- T cells expressing interleukin-5 mRNA. AB - Wells' syndrome, or eosinophilic cellulitis, is a rare dermatosis characterized histologically by a dermal infiltrate of eosinophils, lymphocytes and histiocytes between collagen bundles and amorphous or granular eosinophilic deposits on collagen, constituting flame figures. We report a 54-year-old woman with eosinophilic cellulitis whose peripheral blood showed a marked eosinophilia and a high proportion of CD4+CD7- cells before treatment. Reverse transcriptase polymerase chain reaction revealed that CD4+CD7- cells, but neither CD4+CD7+ nor CD4-CD8+ cells, in the circulating mononuclear cells expressed mRNA for interleukin (IL)-5, the major cytokine involved in eosinophilia. The proportion of CD4+CD7- cells decreased, and expression of mRNA for IL-5 disappeared in the peripheral blood, when the disease was treated by the administration of intravenous recombinant interferon-gamma. These findings suggest that circulating CD4+CD7- T cells play a pivotal role in the pathogenesis of eosinophilic cellulitis by producing IL-5. PMID- 9217825 TI - Muir-Torre syndrome: clinical features and molecular genetic analysis. AB - We report a 62-year-old man with rectal cancer, two keratoacanthomas and multiple sebaceous adenomas, epitheliomas and sebaceous hyperplasia. His brother and father died from colorectal cancer. A subgroup of patients with the Muir-Torre syndrome (MTS) is allelic to the cancer family syndrome. This genetic disorder is caused by an autosomal dominant inherited germline mutation in one of the DNA mismatch repair genes. It is thought that a somatic mutation of the other allele leads to a genomic instability responsible for tumorigenesis. In the patient presented here the instability was detected in two characteristic skin lesions; sebaceous adenoma and epithelioma. The search for a causal germline mutation revealed a frameshift mutation in the mismatch repair gene hMSH2 leading to a truncated protein. A presymptomatic molecular diagnosis can be offered to the children of the patient. PMID- 9217827 TI - Simultaneous onset of primary cutaneous B-cell lymphoma and human herpesvirus 8 associated Kaposi's sarcoma. AB - We report the simultaneous occurrence of Kaposi's sarcoma (KS) and primary cutaneous B-cell lymphoma (CBCL) of the leg in a 79-year-old woman, seronegative for HIV-1, HTLV-1 and HTLV-2. The CBCL underwent complete clinical remission after local radiotherapy, whilst the KS became disseminated within a year following diagnosis. However, 2 years after the diagnosis of KS, the patient died with neurological symptoms. These were presumed to be due to involvement of the central nervous system by lymphoma, although in the absence of an autopsy, this could not be proven. Skin biopsies from the original KS and CBCL lesions, as well as short-term culture of spindle cells from the KS lesion and peripheral blood mononuclear cells (PBMC), were studied by semiquantitative polymerase chain reaction (PCR) using primers specific for DNA sequences of a novel gamma herpesvirus-8 (HHV-8). PCR studies were strongly positive for the virus on KS cells and PBMC; conversely, a low viral load was found on CBCL cells. A high titre of serum IgG antibodies reacting with the nuclei of the HHV-8 positive cell line BCP-1 was found. These data suggest that reactivation of latent infection with HHV-8 had occurred in this patient, and that HHV-8 is directly involved in KS, but not in CBCL of the leg, an aggressive variant of CBCL. PMID- 9217828 TI - Active psoriasis and profound CD4+ lymphocytopenia. AB - We report the case of a patient with a long-standing history of widespread chronic plaque psoriasis, who was recently found to have a profound CD4+ lymphocytopenia. He is human immunodeficiency virus (HIV) negative. His psoriasis remains active and widespread, and he has had 60 cutaneous malignancies, including many squamous cell carcinomas, excised over the last 10 years. In the past he has had numerous cutaneous viral warts. Despite a low peripheral blood CD4+ T-cell count, similar numbers of activated T cells, identified by double labelling for CD4 and HLA-DR antigens, were found in the epidermis of our patient as other individuals with psoriasis. Thus, there appear to be sufficient activated CD4+ T cells in our patient's psoriatic plaques to maintain the psoriatic process. PMID- 9217829 TI - Ectopic respiratory epithelium associated with multiple malformations. AB - We report a patient with unique cutaneous plaques of ectopic respiratory epithelium. The epithelium was located superficially as raised erythematous plaques on the right lateral surface of the neck with some viscous secretion. Underlying branchial cysts or sinuses were excluded. The occurrence of ectopic respiratory epithelium was associated with congenital deafness and a hare-lip in our patient, suggesting multiple malformation during early embryonic development. PMID- 9217830 TI - Olmsted syndrome: report of a new case. AB - We report the case of a 20-year-old man, who was born with an intense erythema of the genital area, unresponsive to any treatment employed. When he was 9 months old, he presented with well-defined hyperkeratotic erythematous plaques around the mouth, eyes, nose, and perianal area, with similar plaques on the lateral aspect of the neck and axillae. At the same time the erythema of the genital area became hyperkeratotic. When he was 2 years old, he presented with a disabling palmoplantar keratoderma, initially focal, and later diffuse, also unresponsive to local or systemic treatments employed. The lesions have varied during the course of the disease without ever clearing completely. The axillary and inguinal plaques have shown spontaneous resolution on occasion. Six skin biopsies have been performed with no conclusive histological diagnosis of any of the typical disorders of keratinization. All treatments, topical and systemic, including etretinate and acitretin, have failed to improve the condition. We believe that this patient has Olmsted syndrome, a rare form of palmoplantar keratoderma with periorificial keratotic plaques. PMID- 9217831 TI - Systemic involvement in scleredema of Buschke associated with IgG-kappa paraproteinaemia. AB - Scleredema is a rare primary cutaneous mucinosis. Systemic involvement is uncommon and histological confirmation is often lacking. We report a case of a 60 year-old man with scleredema and evidence of mucin deposition on biopsies from multiple extracutaneous sites. The bone marrow, nerve, hepatic and salivary gland involvement seen on histology in our patient has not, to our knowledge, been previously reported in this condition. PMID- 9217832 TI - Contact dermatitis with cervical lymphadenopathy following exposure to the hide beetle, Dermestes peruvianus. AB - Contact with beetles of the family Dermestidae can produce a variety of disorders including skin, gastrointestinal and respiratory tract disease. Dermatological disorders include dermatitis, vesicular, pustular and vasculitic lesions. In addition, there may be pruritus, desquamation and urticaria. We report a patient who developed dermatitis, a vasculitic eruption, cervical lymphadenopathy and pulmonary nodular interstitial infiltration as a result of contact with the 'hide beetle' Dermestes peruvianus. PMID- 9217833 TI - Unusual skin ulceration in an HIV-positive patient who had cutaneous syphilis and neurosyphilis. AB - We present a case of a patient coinfected with syphilis and the human immunodeficiency virus (HIV) who had unusual and severe cutaneous ulceration. The profound immune defects associated with HIV may lead to an altered clinical presentation and a more aggressive course in patients infected with Treponema pallidum. Despite non-confirmatory histological findings, we feel our patient's cutaneous ulcers probably represent superficial gummata, which have failed to resolve completely following currently accepted high-dose antisyphilis chemotherapy. PMID- 9217834 TI - Subcutaneous lesions and bacteraemia due to Stenotrophomonas maltophilia in three leukaemic patients with neutropenia. AB - Subcutaneous lesions were seen in three of 13 neutropenia patients who had Stenotrophomonas (Xanthomonas) maltophilia bacteraemia. The characteristic clinical presentation resembled leukaemic infiltrates, and were different from deep ulcers or subcutaneous nodules caused by Pseudomonas aeruginosa. The three patients had acute leukaemia and were treated with intensive combination chemotherapy. All had previously been treated with broad-spectrum antibiotics, and each patient recovered after proper combination antibiotic treatment given according to sensitivity testing. PMID- 9217836 TI - Large patches of Bowen's disease treated by topical aminolaevulinic acid photodynamic therapy. AB - Large patches of Bowen's disease (intraepidermal carcinoma in situ) can be difficult to treat by conventional methods. Photodynamic therapy (PDT) uses the combination of a photosensitizer, which preferentially accumulates in malignant cells, and photoactivation by visible light to kill the malignant cells. 5 aminolaevulinic acid (ALA) PDT uses excess exogenous ALA, which produces, via the haem synthesis pathway, a build up of the photosensitizer protoporphyrin IX. We describe the use of topical ALA PDT to treat three patients with three especially large patches of Bowen's disease. Following two treatments all three lesions achieved a complete clinical and histological response with a good cosmetic result. ALA PDT is a simple, effective and well tolerated treatment for large patches of Bowen's disease. PMID- 9217835 TI - Atypical eumycetoma caused by Phialophora parasitica successfully treated with itraconazole and flucytosine. AB - Phialophora species are occasional pathogens causing subcutaneous and invasive disease. We report the first case of eumycetoma caused by P. parasitica in an otherwise healthy U.K. resident who visited India. She failed to respond to surgical excision and itraconazole, 400 mg daily, but responded to itraconazole, 400 mg daily, and flucytosine, 1 g three times daily, for 12 months. In vitro susceptibility testing predicted a response. PMID- 9217838 TI - Normal recovery of the stratum corneum barrier function following damage induced by tape stripping in patients with atopic dermatitis. AB - Patients with atopic dermatitis (AD) constantly inflict mechanical damage to their skin by scratching induced by pruritus. On excoriated lesions of the cheek we found exceedingly high levels of transepidermal water loss (TEWL) as compared to those in the normal skin of healthy subjects. However, it is not clear whether the skin of patients with AD also shows an abnormally slow recovery after mechanical damage. We compared the recovery of the barrier function of the stratum corneum (SC), after its complete removal by tape stripping, in patients with AD and age-matched healthy control subjects. On the normal-looking skin of the flexor forearm, we found no difference in the recovery process of the water barrier function of the SC between the two groups. This suggests that ability to reconstruct SC barrier function after mechanical damage is not impaired in AD patients. PMID- 9217837 TI - Quantitative assessment of feline epidermal Langerhans cells. AB - The densities of feline epidermal dendritic cells expressing CD18, MHC class II and CD1a antigens were determined for four anatomical locations in 19 cats of European breed in blind conditions. The densities (+/- SD) of CD1a+ Langerhans cells in the skin of the abdominal wall (269 +/- 68 cells/mm2), the back (363 +/- 19), the internal side of the ear (572 +/- 30) and the external side of the ear (502 +/- 32) were significantly different, with young and old animals displaying less stained cells than adults. No significant differences in the mean densities were found with regard to sex, colour or antibody used. PMID- 9217839 TI - Seasonal patterns in the diagnosis of cutaneous malignant melanoma: analysis of the data of the German Central Malignant Melanoma Registry. PMID- 9217840 TI - Validation of the recommendations on clinic size made by the British Association of Dermatologists. PMID- 9217841 TI - Palpebral oedema associated with sensitization to cobalt in a dental prosthesis. PMID- 9217842 TI - Actinic prurigo: limited effect of PUVA. PMID- 9217843 TI - Pharmacodynamic interaction with phototoxic plants during PUVA therapy. PMID- 9217844 TI - Calcium antagonist-induced photo-exposed telangiectasia. PMID- 9217845 TI - Toxic pustuloderma induced by intracavernous prostaglandin E1. PMID- 9217846 TI - Cholestatic jaundice due to terbinafine. PMID- 9217847 TI - Vitamin D receptor polymorphism and treatment of psoriasis with calcipotriol. PMID- 9217848 TI - Granuloma faciale entirely in an extrafacial location. PMID- 9217849 TI - Uveitis in psoriasis associated with HLA-B51; a link to Behcet's disease? PMID- 9217850 TI - Urticaria and hepatitis C. PMID- 9217851 TI - Localized argyria caused by silver earrings. PMID- 9217852 TI - Localised pain following contact with liquids. PMID- 9217853 TI - Neurotic disorders in the elderly: often missed, poorly treated. PMID- 9217854 TI - Intensive therapy of acute ischaemic stroke. PMID- 9217855 TI - Rhinosinusitis. AB - Rhinosinusitis, inflammation of the lining of the nose and paranasal sinuses, has a range of different causes. This article considers the classification, diagnosis and treatment of rhinosinusitis. PMID- 9217856 TI - Update on vaccination guidelines. AB - Updated guidelines for the immunization of children and adults in the UK, incorporating a number of important changes, have recently been published. This article reviews the areas where recommendations have changed significantly and summarizes immunization guidelines in other problem areas. PMID- 9217857 TI - Left ventricular volume reduction for end-stage heart failure: new horizon or false dawn? PMID- 9217858 TI - Keyhole coronary bypass surgery. PMID- 9217859 TI - Dynamic cardiomyoplasty. PMID- 9217860 TI - Long-term implantable circulatory support. PMID- 9217861 TI - Percutaneous tracheostomy: how to do it. AB - Percutaneous tracheostomy has become a commonly used technique in the management of the critically ill. It is a relatively simple procedure to perform at the bedside but it does have, as with any practical procedure, potential pitfalls and complications. Two commonly used methods are described. PMID- 9217862 TI - Rocuronium and cisatracurium. AB - Rocuronium and cisatracurium are the two most recent muscle relaxant additions to our pharmacopocia. These two drugs are significant advances and are likely to have an increasingly important role in clinical anaesthesia in the future. PMID- 9217863 TI - Women doctors: multiprofessional working in health care. AB - In the NHS today, professionals and agencies have to work together to achieve patient focus and integrate care. A barrier to this is the male-dominated culture which endures both in medicine and management. Such work cultures are not only damaging to women doctors, but to health care. There is a need for gender balance at all levels of medicine. PMID- 9217864 TI - Somatic gene therapy. PMID- 9217865 TI - Hypotensive anaesthesia. PMID- 9217866 TI - Patient/doctor relationships. PMID- 9217867 TI - Diabetes mellitus and the cardiovascular system. PMID- 9217868 TI - Actions of insulin on the mammalian heart: metabolism, pathology and biochemical mechanisms. PMID- 9217869 TI - Regulation of energy substrate metabolism in the diabetic heart. AB - The effects of diabetes on myocardial metabolism are complex in that they are tied to the systemic metabolic abnormalities of the disease (hyperglycemia and elevated levels of free fatty acid and ketone bodies), and changes in cardiomyocyte phenotype (e.g., down-regulation of glucose transporters and PDH activity). The cardiac adaptations appear to be driven by the severity of the systemic abnormalities of the disease. The diabetes-induced changes in the plasma milieu and cardiac phenotype both cause impaired glycolysis, pyruvate oxidation, and lactate uptake, and a greater dependency on fatty acids as a source of acetyl CoA. Studies in isolated hearts suggest that therapies aimed at decreasing fatty acid oxidation, or directly stimulating pyruvate oxidation would be of benefit to the diabetic heart during and following myocardial ischemia. PMID- 9217871 TI - Regulation of intracellular Ca2+ in the heart during diabetes. AB - Cardiovascular disease is a significant medical problem. The diabetic population is even more susceptible to cardiovascular complications and heart failure than non-diabetic patients. Atherosclerotic complications, a neuropathy and microvascular lesions have all been implicated causally in the accelerated cardiovascular disease during diabetes. However, one mechanism which may participate in the abnormalities in heart performance demonstrated during diabetes and may also contribute to heart failure in the diabetic is a derangement in the capacity of the myocardial cell to regulate its [Ca2+]. The purpose of this treatise is to identify the current controversies and conclusions available regarding the specific defects in Ca2+ flux thought to contribute to these cardiac defects during diabetes mellitus. PMID- 9217870 TI - Regulation of contractile proteins in diabetic heart. AB - Diabetes is one of the most prevalent chronic conditions that has a high association with death from cardiovascular disease(s). An impaired cardiac function independent of vascular disease suggests the existence of a primary myocardial defect in diabetes mellitus. We and others have documented that myocardial performance is impaired in the hearts of chronically diabetic rats and rabbits. Abnormalities in the contractile proteins and regulatory proteins could be responsible for the mechanical defects in streptozotocin (STZ)-diabetic hearts. The major focus of research on contractile proteins in the diabetic state has been on myosin ATPase and its isoenzymes. However, in the contractile protein system, this could be only one of the mechanisms that might be a controlling factor in myofilament contraction in diabetes. To define the role of cardiac contractile as well as regulatory proteins (troponin-tropomyosin) as a whole in the regulation of actomyosin system in diabetic cardiomyopathy, individual proteins of the cardiac system were reconstituted under controlled conditions. Enzymatic data confirmed a diminished calcium sensitivity in the regulation of the cardiac actomyosin system when regulatory protein(s) complex was recombined from diabetic hearts. This diminished calcium sensitivity along with shifts in cardiac myosin heavy chain (V1-->V3) could contribute to the impaired cardiac function in the hearts of chronic diabetic rats. It has also been reported that sarcomeric proteins such as myosin light chain-2 (MLC-2) and troponin I (TnI) could be involved in regulating muscle contraction and in calcium sensitivity. Since phosphorylation of cardiac TnI is associated with altered maximum enzymatic activity and calcium force relationship in isolated muscle preparations. TnI phosphorylation could contribute to depressed myocardial contractility in experimental diabetes. While we have yet to understand the exact function of each component in cardiac muscle and their behavior in concert where all of them act in tandem, we have focussed on the role of contractile proteins and their regulation in diabetes in this review. We have also included a brief discussions on other relevant intracellular components. In summary, there is substantial evidence to suggest that there are independent processes associated with diabetes which effect cardiac performance in experimental animals and in man. The focus of this review has been the explication of a biochemical defect which underlies cardiac contractile dysfunction in experimental models of diabetes. PMID- 9217872 TI - The regulation of intracellular pH in the diabetic myocardium. PMID- 9217873 TI - Endothelial dysfunction and pathogenesis of diabetic angiopathy. AB - OBJECTIVE AND METHODS: To review, from the clinical perspective, the contribution of dysfunction of the vascular endothelium to the pathogenesis of diabetic micro- and macroangiopathy. RESULTS: Available data indicate that endothelial dysfunction in diabetes complicated by micro- or macroalbuminuria (renal microangiopathy) is generalised. The close linkage between microalbuminuria and endothelial dysfunction is an attractive explanation for the fact that microalbuminuria is a risk marker for atherosclerotic cardiovascular disease in diabetes. Endothelial dysfunction precedes the occurrence of even early diabetic microangiopathy. However, it is not clear whether endothelial dysfunction is a feature of the diabetic state per se or whether additional factors are required to induce endothelial dysfunction given the presence of diabetes. Convincing data from animal and in vitro models exist to indicate that endothelial dysfunction in diabetes may be related to hyperglycaemic pseudohypoxia, activation of protein kinase C, increased expression of transforming growth factor-beta and vascular endothelial growth factor, non-enzymatic glycation, oxidative stress, activation of the coagulation cascade, increased expression of tumour necrosis factor-alpha, and high levels of insulin and insulin precursor molecules. However, the importance of these proposed mechanisms have not yet been extensively assessed in diabetes in man. CONCLUSIONS: Endothelial dysfunction plays a key role in the pathogenesis of diabetic angiopathy in man. The biochemical basis of endothelial dysfunction in diabetic man, however, has yet to be fully elucidated. PMID- 9217874 TI - Insulin secretion and its modulation by antiarrhythmic and sulfonylurea drugs. AB - Cardiovascular drugs such as antiarrhythmic agents with Vaughan Williams class Ia action have been found to induce a sporadic hypoglycemia. Recent investigation has revealed that these drugs induce insulin secretion from pancreatic beta-cells by inhibiting ATP-sensitive K+ (KATP) channels in a manner similar to sulfonylurea drugs. The mechanism underlying block of KATP channels by antiarrhythmic drugs was different, however, from that of sulfonylureas: firstly, because binding of radioactive glibenclamide could not be inhibited by unlabelled antiarrhythmic agents, and vice versa; secondly, because the two compounds differ in the kinetics and sidedness of drug action-antiarrhythmic drugs act on the channel from the inner surface of the cell membrane, whereas glibenclamide binds through the intramembrane pathway; finally, it was shown that functional KATP channels in beta-cells are composed of two distinct molecules-a sulfonylurea receptor (SUR) and a channel pore-forming subunit, an inwardly-rectifying K channel with two transmembrane regions (Kir6.2). Antiarrhythmic drugs reversibly inhibit the K+ conductance displayed by the Kir6.1 (a putative KATP channel clone)-transfected NIH3T3 cells. Therefore they appear to interact directly with the pore-forming subunit, thereby inhibiting KATP channel currents and exerting an insulinotrophic effect. PMID- 9217875 TI - Sulfonylurea derivatives in cardiovascular research and in cardiovascular patients. AB - Sulfonylurea derivatives are hypoglycemic drugs frequently used in the treatment of non-insulin-dependent diabetes mellitus (NIDDM). In the beta-cell sulfonylureas act by blocking ATP-sensitive potassium channels (K.ATP channels). In several organ systems, including the cardiovascular system, sulfonylurea receptors and functional K.ATP channels have been identified. In the heart their role is not clear: an endogenous cardioprotective effect has been suggested. There is no doubt that K.ATP channels are effectively blocked by sulfonylureas. In the last decade sulfonylureas have been widely used as a pharmacological tool in experimental (cardiac) research. Blockade of K.ATP channels is the proposed cellular mechanism of action for all sulfonylurea-related effects. However, other membrane currents are affected as well. In addition, myocardial metabolism is modified by sulfonylurea pretreatment. Hence, it should seriously be questioned whether these drugs are suitable in assessing involvement of cardiac K.ATP channels in, for example, ischemia-related events. The detrimental effects of sulfonylureas in experimental studies on myocardial ischemia have led to speculation whether the widespread use of these drugs in patients with NIDDM, most often suffering from accompanying ischemic heart disease, should be reconsidered. However, a review of the clinical literature reveals that the most consistent finding is a lower incidence of ventricular arrhythmias associated with the use of glibenclamide, while no excess mortality has been shown for this agent in NIDDM with ischemic heart disease. Despite some direct effects on systemic and coronary vasculature, there are, at present, no firm clinical data on the basis of which sulfonylurea derivatives should be withheld from the cardiac patient. PMID- 9217876 TI - Volume reflex in diabetes. AB - After examining various components of the volume reflex in different models of diabetic rats, it is clear that the neural component of the volume reflex is altered in the early stage of diabetes. Nevertheless, the mechanisms involved in the neural component need to be examined further in diabetes. In terms of the humoral component of the effector limb of the volume reflex, the actions of ANF within the kidney appear to contribute to the altered volume reflex in diabetes. Results of our preliminary studies suggest that intrarenal factors may also contribute to the blunted renal excretory responses to acute VE in early diabetes. Finally, the present review has outlined the major abnormalities in the volume reflex in diabetes and the gaps in our knowledge of the various components of the volume reflex in diabetes. PMID- 9217877 TI - Intracellular calcium levels are unchanged in the diabetic heart. AB - The quality control indices of myocyte isolation (viability, yield, survival time, cell response, etc.) suggest that the adult rat myocyte model is stable and useful in [Ca2+]i measurements and functional studies at the cellular level. Moreover, diabetic cardiomyocytes are a valuable model for studying cellular functions of the diabetic heart as they retain most of the features of cardiac dysfunction of intact rat. Data from our studies indicate that the basal [Ca2+]i in both quiescent and electrically-stimulated cells is not changed. Thus, resting levels of [Ca2+]i and basal [Ca2+]i transients may not reflect the abnormalities observed in diabetes until the system is challenged by certain stimuli. [Ca2+]i responses to isoproterenol are depressed in both resting and stimulated diabetic cells. This suggests an alteration in the beta-adrenergic pathway, possibly related to the beta-adrenoceptor deficiency reported in the diabetic heart. SR Ca ATPase is also involved in the isoproterenol-induced [Ca2+]i changes. Moreover, the decreased maximum response to 8-bromo-cAMP provides evidence of a post receptor alteration in the pathway. Diabetic myocytes are more sensitive to ouabain, whereas the maximum response to ouabain was depressed. This may be the result of depressed Na-K ATPase and increased [Na+]i. In diabetic myocytes, rapid cooling contractures and caffeine contractures are depressed, whereas caffeine induced Ca2+ transients are decreased. Ryanodine binding suggests a decreased number of high-affinity binding sites in the SR of diabetic myocytes. Additionally, there are indications that SR releasable calcium is reduced and that the major functions of SR, notably uptake, release and storage, may be depressed in diabetic myocytes. Finally, L-type Ca(2+)-channels are quantitatively and qualitatively altered in diabetes. Insulin treatment normalizes most of the diabetes-induced changes in cardiomyocytes, suggesting that metabolic alterations due to insulin deficiency play an important role in diabetic cardiomyopathy. Results from several studies show that in diabetes the function of major organelles which handle [Ca2+]i in myocytes is depressed, which in turn causes the alteration of [Ca2+]i mobilization in myocytes. Different second messenger systems involved in E-C coupling may also be altered due to the metabolic impairments. The rapid increase in our understanding of the pathophysiology of calcium homeostasis in cardiomyocytes will be forthcoming as the powerful new tools of molecular and structural biology are used to investigate the regulation of the Ca2+ transport system. PMID- 9217878 TI - Cytosolic Ca2+ concentration decreases in diabetic rat myocytes. PMID- 9217879 TI - The diabetic heart is more sensitive to ischemic injury. AB - Clinical studies have suggested that the diabetic heart is more sensitive to ischemic injury than the non-diabetic heart. However, results from a number of experimental studies using animal models of diabetes reported no change, increased or decreased sensitivity to ischemia. The purpose of this review is to discuss the possible explanations for this apparent discrepancy. Analysis of the conflicting literature on this subject reveals a pattern which suggests that the disparity of experimental findings stems from differences in the duration and severity of the diabetic state, the ischemic flow rate and whether fatty acids are provided as an exogenous substrate. It appears that short-term or mild diabetes is associated with decreased sensitivity to zero-flow ischemic injury. However, as the duration or severity of diabetes increases, this beneficial effect disappears. The diabetic heart also appears to be more vulnerable to injury during low-flow ischemia and when elevated fatty acids are present. PMID- 9217880 TI - Controversies on the sensitivity of the diabetic heart to ischemic injury: the sensitivity of the diabetic heart to ischemic injury is decreased. AB - Controversy exists as to whether the diabetic heart is more or less sensitive to ischemic injury. Although a considerable number of experimental studies have directly determined the effects of ischemia on the diabetic heart, there is still no general agreement as to whether metabolic changes within the myocardium contribute to the severity of ischemic injury. This paper reviews the evidence suggesting that the diabetic heart can actually be less sensitive to an episode of severe ischemia. Possible reasons for this decreased sensitivity to injury are discussed, which include a decreased accumulation of glycolytic products during ischemia (lactate and protons), as well as alterations in the regulation of intracellular pH in the diabetic heart. Based on existing studies, we suggest that although impaired glucose metabolism in the diabetic heart contributes to injury in hypoxic hearts or in hearts subjected to low-flow ischemia, diabetes induced decreases in glycolysis can actually be beneficial to the diabetic heart during and following a severe ischemic episode. A decreased clearance of protons via the Na+/H+ exchanger may also contribute to the decreased sensitivity to ischemic injury in the diabetic heart. PMID- 9217881 TI - Chronic T-type Ca2+ channel blockade with mibefradil in hyperinsulinemic, insulin resistant and hypertensive rats. AB - OBJECTIVES: To determine the effects of calcium antagonists on hyperinsulinemia, hypertriglyceridemia and hypertension, we examined the long-term effects of a new calcium channel blocker, mibefradil, on plasma insulin levels, plasma triglyceride levels and systolic blood pressure in insulin-resistant and hyperinsulinemic fructose-hypertensive (FH) rats. To this aim, both prevention and reversal protocols were employed. METHODS: Prevention study: Male Sprague Dawley rats were procured at 6 weeks of age and were divided into: control (C, n = 6), control-treated (CT, n = 5), fructose (F, n = 7) and fructose-treated (FT, n = 6). Baseline measurements of plasma glucose, insulin and systolic blood pressure were conducted in all groups. At week 7, chronic mibefradil treatment (30 mg/kg/day, orally for 6 weeks) was initiated in the CT and FT groups. At week 8, the rats in the F and FT groups were started on a 66% fructose diet to induce hyperinsulinemia and hypertension. Weekly measurements of plasma insulin, plasma triglycerides and systolic blood pressure were conducted for the following 4 weeks. Reversal protocol: In a separate study, 8-week-treated FH rats and their age-matched controls were used to examine the effects of mibefradil on reversing fructose-induced hyperinsulinemia and hypertension. RESULTS: The F group exhibited hyperinsulinemia (3.2 +/- 0.1 vs. C 2.3 +/- 0.07 ng/ml, P < 0.05), hypertension (148 +/- 3 vs. C 121 +/- 1 mmHg, P < 0.002) and elevated triglyceride levels (5.4 +/- 0.8 vs. C 1.6 +/- 0.3 mM, P < 0.05). Chronic mibefradil treatment prevented the development of hyperinsulinemia (1.6 +/- 0.08 ng/ml, P < 0.004 vs. F) and hypertension (123 +/- 1 mmHg. P < 0.001 vs. F) and attenuated the development of hypertriglyceridemia. In the reversal study, mibefradil treatment reversed the development of hyperinsulinemia, hypertriglyceridemia and elevated BP in FH rats. Treatment did not affect the plasma glucose levels in any group (prevention or reversal). CONCLUSIONS: Long term treatment with the calcium antagonist, mibefradil, both prevents and reverses the development of hyperinsulinemia, hypertriglyceridemia and hypertension in FH rats. These data indicate beneficial effects of mibefradil on carbohydrate and lipid metabolism in hyperinsulinemic and insulin-resistant states. PMID- 9217882 TI - Mechanisms underlying depressed Na+/Ca2+ exchanger activity in the diabetic heart. AB - OBJECTIVES: Depression in Na+/Ca2+ exchanger activity is an important factor in the development of the diabetic cardiomyopathy. Since the mechanism underlying this depression remains unknown, the aim of this study was to determine the contribution of hyperglycemia and insulinopenia towards the observed impairment in Na+/Ca2+ exchanger activity. METHODS: Non-insulin-dependent diabetes was induced in neonatal Wistar rats by injection of 90 mg/kg streptozotocin. Na+/Ca2+ exchange in sarcolemmal vesicles and isolated cardiomyocytes was determined by Na(+)-dependent 45Ca2+ transport. To assess the role of insulin deficiency and hyperglycemia on Na+/Ca2+ exchanger activity, neonatal cardiomyocytes were incubated for 3 days in media containing either 5 mM glucose and 56 U/l insulin (Control), 30 mM glucose and 56 U/l insulin (High glucose) or 5 mM glucose and 0 insulin (Insulin deficiency). Since hyperglycemia has been shown to affect protein kinase C activity, Ca(2+)-dependent isoforms of protein kinase C were examined in non-diabetic and diabetic heart using hydroxylapatite chromatography. Also examined was Na+/Ca2+ exchanger mRNA levels in diabetic and non-diabetic hearts using Northern slot blot analysis. RESULTS: Acute insulin produced a dose dependent increase in Na+/Ca2+ exchanger activity, which was dramatically attenuated in diabetic membrane. Myocytes incubated in media containing 30 mM glucose exhibited a 33% reduction in Na+/Ca2+ exchanger activity, while insulinopenia reduced activity by 63%. Exchanger mRNA levels of the diabetic heart were normal; however, diabetes was associated with major changes in protein kinase C activity. CONCLUSIONS: Reduced Na+/Ca2+ exchanger activity resulting from diabetes, hyperglycemia or insulinopenia may be related to changes in protein kinase C activity, but is not caused by altered expression of the transporter. PMID- 9217883 TI - Na+/K(+)-ATPase activity in vascular smooth muscle from streptozotocin diabetic rat. AB - OBJECTIVES: Insulin-deficient diabetes impairs carbohydrate metabolism in a variety of tissues. Vascular smooth muscle may be susceptible to the diabetes induced disturbance in glycolysis since Na+/K(+)-ATPase in this tissue preferentially utilizes ATP generated by glycolysis. The purpose of this study was to determine if chronic exposure to the metabolic alterations associated with insulin-deficient diabetes directly inhibited Na+/K(+)-ATPase activity, or its regulation, in vascular smooth muscle. METHODS: Diabetes was induced by intravenous administration of streptozotocin (60 mg/kg). After 12 weeks, Na+/K(+) ATPase activity in aorta and superior mesenteric artery was evaluated under a variety of conditions. Na+/K(+)-ATPase was estimated by measuring the influx of rubidium-86 (86Rb) in the presence or absence of the Na+/K(+)-ATPase inhibitor, ouabain. The metabolism of [3H]glucose and [14C]glucose was used to estimate glycolysis or glucose oxidation, respectively. RESULTS: Glycolysis and glucose oxidation were decreased in aortic smooth muscle (27 and 34%, respectively). An intact endothelium was associated with a marked decrease in ouabain-sensitive (pump-mediated) 86Rb uptake in diabetic aorta. However, ouabain-sensitive 86Rb uptake was similar in de-endothelialized aorta and superior mesenteric artery from diabetic and non-diabetic rats under both unstimulated conditions and during maximal stimulation. Removal of glucose or oxygen reduced ouabain-sensitive 86Rb uptake to a similar extent in both groups. In contrast, the receptor-mediated stimulation of ouabain-sensitive 86Rb uptake by insulin was decreased. CONCLUSIONS: These results suggest that intrinsic Na+/K(+)-ATPase activity is not diminished in diabetic vascular smooth muscle under physiological conditions and that the impairment of cellular metabolism in diabetic blood vessels does not limit stimulation of Na+/K(+)-ATPase activity. However, modulation of Na+/K(+) ATPase activity by endothelial factors or insulin appears to be altered in aorta from diabetic rats. PMID- 9217884 TI - Diabetic-induced endothelial dysfunction in rat aorta: role of hydroxyl radicals. AB - OBJECTIVE: Previous studies suggest a role of superoxide anion radicals (.O2-) in impaired endothelium-dependent relaxation of diabetic blood vessels; however, the role of secondary reactive oxygen species remains unclear. In the present study, we investigated a role of various potential reactive oxygen species in diabetic endothelial dysfunction. METHODS: Thoracic aortic rings from 8-week streptozotocin-induced diabetic and age-matched control rats were mounted in isolated tissue baths. Endothelium-dependent relaxation to acetylcholine (ACH) and endothelium-independent relaxation to nitroglycerin (NTG) were assessed in precontracted rings. RESULTS: ACH-induced relaxation was impaired in diabetic compared to control rings and was not improved with either indomethacin or daltroban. ACH-induced relaxation in both control and diabetic rings was completely blocked with the nitric oxide synthase inhibitors, L-nitroarginine methyl ester or L-nitroarginine (L-NA). NTG-induced relaxation was insensitive to L-NA and was unaltered by diabetes. Pretreatment with superoxide dismutase (SOD) at activities which did not alter contractile tone failed to alter response to ACH in diabetic rings. Similar results were obtained using either catalase or mannitol. In contrast, the combination of SOD plus catalase or DETAPAC, an inhibitor of metal-facilitated hydroxyl radical (.OH) formation, markedly enhanced relaxation to ACH in diabetic but not in control rings. Neither the combination of SOD plus catalase nor DETAPAC altered the sensitivity or relaxation to NTG in control rings with or without endothelium. In diabetic rings with endothelium, both DETAPAC or SOD plus catalase increased sensitivity but not maximum relaxation to NTG. In diabetic rings without endothelium, relaxation and sensitivity to NTG were unaltered by either treatment. In L-NA-treated diabetic rings with endothelium, sensitivity and relaxation to NTG was unaltered by either DETAPAC or SOD plus catalase. CONCLUSION: Diabetic endothelium produces increases in both .O2- and H2O2 leading to enhanced intracellular production of .OH. Thus, .OH are implicated in diabetes-induced endothelial dysfunction. PMID- 9217886 TI - Endothelial dysfunction of conduit arteries in insulin-dependent diabetes mellitus without microalbuminuria. AB - OBJECTIVE: Previous studies have shown that endothelial dysfunction, an early sign of atherosclerosis, occurs in animal models of diabetes mellitus and in resistance vessels of patients with insulin-dependent diabetes. In the present study we examined whether young patients with insulin-dependent diabetes without microalbuminuria present abnormal endothelial function of large peripheral arteries. METHODS: Twenty-six patients with insulin-dependent diabetes without microalbuminuria were compared with 26 normal controls and 5 patients with insulin-dependent diabetes with microalbuminuria. Brachial artery diameter was measured at rest, during reactive hyperaemic flow (endothelium-dependent dilatation) and after sublingual isosorbide dinitrate (endothelium-independent dilatation). RESULTS: Baseline artery diameter and flow as well as the degree of reactive hyperaemia were similar in all groups compared to controls. Flow mediated dilatation was lower in patients with diabetes without microalbuminuria (5.8 +/- 7 vs 11 +/- 7%. P = 0.01) as well as in patients with diabetes without microalbuminuria (0.75 +/- 2.5 vs 11 +/- 7%, P = 0.003); nitrate-induced dilatation was normal in patients without microalbuminuria and attenuated in patients with microalbuminuria. In the group of diabetes patients without microalbuminuria, those with disease duration > 10 years and HbAlc > 6% had the worse endothelial function. CONCLUSIONS: Our results demonstrate that endothelial dysfunction of conduit arteries can be detected in patients with insulin dependent diabetes mellitus without microalbuminuria, probably contributing to the high prevalence of atherosclerosis in these patients. PMID- 9217885 TI - Characteristics of coronary endothelial dysfunction in experimental diabetes. AB - OBJECTIVE: To study the influence of diabetes on the endothelium-dependent vasodilation in the coronary arterial bed. METHODS: The effects of acetylcholine (ACh 2-36 pmol.kg-1; 18 nmol.1(-1)-9.8 mumol.1(-1); 0.1-10 mumol.1(-1), L arginine (1 mmol.1(-1) and sodium nitroprusside (1 nmol.1(-1)-100 mumol.1(-1)) were measured on coronary conductivity, vascular tone and cGMP release (RIA) in healthy and diabetic dogs. RESULTS: ACh-mediated (in cumulative intra-arterial infusion) increase in coronary conductivity was reduced (P < 0.01) in the diabetic dogs in vivo, whereas no increase in cGMP release was observed in isolated diabetic coronaries (P < 0.05) which could not be enhanced by L-arginine (P < 0.05). Inhibition of cyclo-oxygenase after 20 min further impaired (P < 0.01) responsiveness to ACh in vivo and diminished the ACh response in isolated coronary strips of the diabetic dogs, but not in those of the controls. Relaxation in response to sodium nitroprusside was not altered by diabetes. CONCLUSIONS: Diminished vasodilation in diabetes is due to a defect in endothelial nitric oxide production and action. Vasodilating prostanoids do not sufficiently compensate this defect. PMID- 9217887 TI - Glucose elevations alter bradykinin-stimulated intracellular calcium accumulation in cultured endothelial cells. AB - OBJECTIVE: Diabetes selectively injures receptor-mediated endothelium-dependent relaxation. In this study, we investigated the effect of elevated glucose concentrations on intracellular calcium (Ca2+i) signal transduction in response to stimulants of EDRF/nitric oxide release in cultured bovine aortic endothelial cells. METHODS: [Ca2+i] was measured in cell suspensions using Fura-2 and fluorescence spectroscopy while nitric oxide production was evaluated using radioimmunoassay of cGMP production. RESULTS: After 24 h exposure to 25 mM glucose in Ham's F-12 media, the increase in endothelial cell [Ca2+i] in response to 100 nM bradykinin was attenuated by 40% while the response to ionomycin was unaltered. When RMPI medium was used, no reduction in response to bradykinin was observed at 25 mM glucose, but a significant reduction in [Ca2+i] signal was observed after exposure to 35 mM glucose for a similar time period. Defective [Ca2+i] signaling was also seen in cells using MEM medium. [Ca2+i] signal responses to ionomycin and NaF, a G-protein activator of extracellular calcium entry via calcium channels, were unaltered by elevated glucose exposure. The defect in [Ca2+i] signal was not mimicked by either mannose or sucrose, but was prevented by co-incubation with cytochalasin B to inhibit glucose uptake. Neither superoxide dismutase nor catalase nor the extracellular hydroxyl radical scavenger, mannitol, blocked the reduction in the bradykinin-induced increase of [Ca2+i] in elevated glucose-exposed cells; however, the reduction was completely blocked by the cell-permeable hydroxyl radical scavenger, dimethylthiourea. Bradykinin-stimulated (but not ionomycin-stimulated) cGMP production within endothelial cells or in RFL-6 detector cells was attenuated by elevated glucose exposure. CONCLUSIONS: Hyperglycemia may contribute to defective endothelium dependent relaxation in diabetes via an attenuated increase in Ca2+i signal transduction for the release of nitric oxide by endothelial cells. This defect possibly arises as a consequence of hydroxyl radicals formed intracellularly. PMID- 9217888 TI - Effects of glycosylated hemoglobin on vascular responses in vitro. AB - Vascular responses to endothelium-dependent vasodilators are greatly impaired in vivo, while isolated blood vessels from animals with diabetes mellitus demonstrate less consistent degrees of impairment. Glycation of proteins, such as hemoglobin, has been implicated in the vascular abnormalities associated with diabetes. OBJECTIVE: The purpose of this study was to test the hypothesis that glycosylated hemoglobin is capable of reducing endothelium-dependent vasodilator responses, possibly explaining impaired dilation observed in vivo. METHODS: To test this hypothesis, the effect of glycosylated hemoglobin (GH) on vascular responses was studied in several vascular beds, including ventricular microvessels and coronary, mesenteric, femoral, and renal arteries. Coronary arterioles were isolated and mounted between two glass pipettes in a pressurized (30 cmH2O) organ chamber. Isolated artery segments were studied using a standard isometric ring technique. RESULTS: In ventricular microvessels, 10 nM nGH (non GH) and GH both attenuated the relaxation to Ach. A lower concentration, 1 nM nGH or GH, did not alter dilation to Ach. In coronary, femoral, mesenteric and renal artery segments, endothelium-dependent responses were not altered by the presence of 10 or 100 nM nGH or GH. CONCLUSION: In coronary microvessels, and coronary, femoral, mesenteric and renal arteries, GH is not responsible for the impaired endothelial function associated with diabetes mellitus. PMID- 9217889 TI - Reduction of 0.1 Hz microcirculatory fluctuations as evidence of sympathetic dysfunction in insulin-dependent diabetes. AB - OBJECTIVE: Loss of spontaneous fluctuations in resting microcirculatory flow has been described in diabetes mellitus, but its mechanism remains unexplained. METHODS: The autonomic control of forearm skin microcirculation was investigated in 23 insulin-dependent diabetic human subjects (median age 39 years, range 27 50) and in 23 age-matched controls (median age 38 years, range 20-57), by laser Doppler flowmetry. Using spectral analysis of spontaneous microvascular fluctuations, we measured the power of 0.1 Hz ('10-second rhythm') fluctuations, dependent on sympathetic control, and of respiration-related, high-frequency fluctuations, due to the transmission of mechanical chest activity. Autonomic function abnormalities were assessed by 5 tests of cardiovascular reflexes. RESULTS: Abnormalities in cardiovascular autonomic tests were present in 7/23 patients: deep breathing was abnormal 4 in patients, standing in 2, handgrip in 3, cross-correlation in 4, and Valsalva ratio in 0. The power of 0.1 Hz microcirculatory fluctuations was significantly lower in diabetic than in control subjects (2.57 +/- 0.16 vs 3.48 +/- 0.09 In-mV2, mean +/- s.e.m., P < 0.001), whereas that of respiratory fluctuations was similar (2.60 +/- 0.24 vs 2.56 +/- 0.19 In-mV2, P = n.s.). The 0.1 Hz power was 2 standard deviations below the mean of controls (P < 0.05) in 13/23 diabetic patients; this abnormality was significantly more frequent than abnormalities in any other autonomic test (P < 0.001). CONCLUSIONS: Since the observed reduction was confined to those microvascular fluctuations under autonomic control, but not to those dependent on passive mechanical transmission, the reduction in spontaneous microcirculatory vasomotion appears to be determined mainly by sympathetic dysfunction. Sympathetic impairment of skin microvascular control seems to be a common finding, and is probably an early index of autonomic dysfunction in insulin dependent diabetes. PMID- 9217890 TI - Peripheral blood flow and noradrenaline responsiveness: the effect of physiological hyperinsulinemia. AB - OBJECTIVE: Insulin seems to have vasodilator properties, but it is unclear if insulin in postprandial concentrations is a specific vasodilator of skeletal muscle resistance arterioles only or that various types of vessels are affected. The aim of the present study was to determine the direct effects and the time course of regional/local physiological hyperinsulinemia on skeletal muscle arterioles, skin blood flow and peripheral venous tone and the responsiveness of these different vascular beds to noradrenaline. METHODS: In protocol I (n = 12) we infused insulin into the brachial artery for 180 min (3.5 mU/min) and evaluated the effects on forearm (muscle) blood flow (FBF) and skin blood flow (SBF). Furthermore, noradrenaline (0.025, 0.01 and 0.4 microgram/min) was infused (i.a.) at baseline, at 90 and 180 min after the start of insulin. In protocol 2 (n = 10) the same regional forearm hyperinsulinemia was achieved, but now the local venous responsiveness to noradrenaline (1.7-55 ng/min, at baseline and at 90 and 180 min) was measured in a dorsal hand vein. In protocol 3 we evaluated the local effects of different doses of insulin (1-100 mU/min) infused directly into hand veins preconstricted with phenylephrine. RESULTS: Forearm hyperinsulinemia (approximately 50 mU/l) led to a significant increase in FBF after 180 min (median 26%, interq ranges 5-50, P < 0.05), while SBF was not altered. Forearm hyperinsulinemia did not affect the noradrenergic responsiveness in skeletal muscle or skin. Infused locally into hand veins only the highest dose of insulin (100 mU/min) caused a minor venodilation (7% [2.4-12.5], P < 0.05). CONCLUSION: Regional forearm physiological hyperinsulinemia has a vasodilator effect on resistance vessels in skeletal muscle, but is slow in onset (180 min). However, skin vasculature and peripheral veins are not affected by this hyperinsulinemia. PMID- 9217891 TI - Strain differences in susceptibility to streptozotocin-induced diabetes: effects on hypertriglyceridemia and cardiomyopathy. AB - OBJECTIVE: Streptozotocin (STZ)-induced diabetes in Wistar rats results in severe hyperlipidemia and a characteristic cardiomyopathy. However, Wistar-Kyoto (WKY) rats made diabetic with a similar dose of STZ did not develop heart dysfunction or hypertriglyceridemia at 12 weeks post-STZ. We investigated whether an apparent resistance of the WKY strain to develop diabetic cardiomyopathy and hypertriglyceridemia following chronic diabetes could be due to a reduced susceptibility to the diabetogenic effects of STZ. METHODS: Adult male WKY and Wistar rats were made diabetic with a moderate (55 mg/kg) or high (75 mg/kg) dose of STZ. At 6 weeks of diabetes, glucose tolerance, cardiac function, pancreatic insulin content and basal and post-heparin plasma lipolytic activity were determined. RESULTS: Administration of a moderate dose of STZ produced cardiac dysfunction in Wistar but not WKY rats at 6 weeks after diabetes induction. The same dose of STZ in WKY rats also resulted in a lesser degree of hyperglycemia and glucose intolerance, and significantly higher pancreatic insulin content relative to Wistar rats. Following a high dose of STZ, the apparent resistance to developing cardiomyopathy was lost in the WKY rats. As well, the WKY rats demonstrated an equal degree of hyperglycemia and glucose intolerance as Wistar rats. However, unlike the Wistar strain, WKY rats did not demonstrate either hypertriglyceridemia or a reduced heparin-releasable plasma lipoprotein lipase (LPL) activity following a high dose of STZ. CONCLUSIONS: These results suggest that the incidence of diabetes-related cardiomyopathy and hypertriglyceridemia in rats may be independently influenced by strain-dependent susceptibilities to the beta-cytotoxic effects of STZ. The absence of hypertriglyceridemia in severely diabetic WKY rats may be linked to the maintenance of a critical level of plasma LPL activity. PMID- 9217892 TI - Effects of physical training on heart rate variability in diabetic patients with various degrees of cardiovascular autonomic neuropathy. AB - OBJECTIVE: To investigate the effects of regularly performed endurance training on heart rate variability in diabetic patients with different degrees of cardiovascular autonomic neuropathy (CAN). METHODS: Bicycle ergometer training (12 weeks, 2 x 30 min/week, with 65% of maximal performance) was performed by 22 insulin-requiring diabetic patients (age 49.5 +/- 8.7 years; diabetes duration 18.6 +/- 10.6 years; BMI 25.1 +/- 3.4 kg/m2): i.e., by 8 subjects with no CAN, 8 with early CAN and by 6 patients with definite/severe CAN. A standard battery of cardiovascular reflex tests was used for grading of CAN, a short-term spectral analysis of heart rate variability for follow-up monitoring of training-induced effects. RESULTS: While the training-free interval induced no changes in spectral indices, the 12-week training period increased the cumulative spectral power of the total frequency band (P = 0.04) but to a different extent (P = 0.039) in different degrees of neuropathy. In patients with no CAN the spectral power in the high-frequency (HF) band (0.15-0.50 Hz) increased from 6.2 +/- 0.3 to 6.6 +/- 0.4 In [ms2]; P = 0.016, and in the low-frequency (LF) band (0.06-0.13 Hz) from 7.1 +/- 0.1 to 7.6 +/- 0.3 in [ms2]; P = 0.08 which resulted in an increase of total spectral power (0.06-0.50 Hz) from 7.5 +/- 0.1 to 8.0 +/- 0.3 in [ms2] (P = 0.05). Patients with the early form of CAN showed an increase of spectral power in HF (5.1 +/- 0.2 to 5.8 +/- 0.1 in [ms2], P = 0.05) and LF bands (5.6 +/- 0.1 to 6.3 +/- 0.1 in [ms2], P = 0.008), resulting in an increase of total power from 6.1 +/- 0.1 to 6.6 +/- 0.1 in [ms2] (P = 0.04), whereas those with definite/severe CAN showed no changes after the training period. Training improved fitness in the whole patient cohort. The increased autonomic tone as assessed by spectral indices disappeared after a training withdrawal period of 6 weeks. CONCLUSIONS: In diabetic patients with no or early CAN, regularly performed endurance training increased heart rate variability due to improved sympathetic and parasympathetic supply, whereas in subjects with definite/severe CAN no effect on heart rate variability could be demonstrated after this kind of training. PMID- 9217894 TI - Prostaglandins and nitric oxide mediate insulin-induced vasodilation in the human forearm. AB - OBJECTIVE: To determine the involvement of prostaglandins, nitric oxide, and beta adrenoceptor activation in insulin-induced vasodilation in the human forearm. METHODS: Fifteen normal subjects were studied. Insulin was administered into the brachial artery in the presence of saline, the cyclo-oxygenase inhibitor indomethacin (0.65 microgram/kg/min), the inhibitor of nitric oxide synthase NG mono-methyl-L-arginine (L-NMMA, 30 micrograms/kg/min), or the non-selective beta adrenoceptor blocker propranolol (0.2 microgram/kg/min). Forearm vascular resistance (FVR) was derived from forearm blood flow and concomitantly measured intra-arterial blood pressure. RESULTS: Insulin decreased FVR by 32 +/- 5% (P < 0.01). Both indomethacin and L-NMMA inhibited insulin-induced vasodilation, while propranolol had no effect. Single infusion of indomethacin was without effect on vascular tone, while single infusion of L-NMMA increased FVR by 81 +/- 19% (P < 0.01). CONCLUSIONS: Insulin has vasodilating properties in skeletal muscle vasculature that is mediated by increases in nitric oxide, that subsequently stimulates prostaglandin release. The latter appears to be a novel vascular action of insulin. PMID- 9217893 TI - L-arginine does not reverse impaired agonist-induced increases in macromolecular efflux during diabetes mellitus. AB - OBJECTIVE: The first goal of this study was to determine the effect of diabetes mellitus on agonist-induced increases in venular macromolecular permeability of the hamster cheek pouch. The second goal was examine the role for an alteration in the availability of L-arginine to nitric oxide synthase in impaired agonist induced increases in macromolecular permeability during diabetes. METHODS: We used intravital fluorescent microscopy and fluorescein isothiocyanate dextran (FITC-dextran; mw = 70 K) to examine macromolecular extravasation from post capillary venules in non-diabetic and diabetic (2 weeks after injection of streptozotocin) hamsters in response to histamine and substance P. Increases in extravasation of macromolecules were quantitated by counting the number of venular leaky sites and by calculating the clearance (ml/s x 10(-6)) of FITC dextran-70 K. RESULTS: In non-diabetic hamsters, histamine (1.0 and 5.0 microM) and substance P (50 and 100 nM) increased permeability of the cheek pouch microcirculation to FITC-dextran-70 K. In contrast, histamine- and substance P induced increases in macromolecular extravasation were markedly reduced in diabetic hamsters. Next, we investigated whether alterations in histamine- and substance P-induced changes in macromolecular extravasation in diabetic hamsters may be related to an alteration in the availability of L-arginine. We examined whether exogenous application of L-arginine (100 microM) could restore impaired histamine- and substance-P-induced increases in macromolecular extravasation in diabetic hamsters. We found that L-arginine potentiated agonist-induced increases in macromolecular extravasation in non-diabetic hamsters, but did not alter responses in diabetic hamsters. CONCLUSION: These findings suggest that short term diabetes mellitus alters agonist-induced alterations in microvascular permeability. The mechanism of altered microvascular permeability during diabetes mellitus does not appear to be related to an impaired availability of L-arginine. PMID- 9217895 TI - Prolonged ejection duration helps to maintain pump performance of the renal hypertensive-diabetic rat heart: correlations between isolated papillary muscle function and ventricular performance in situ. AB - OBJECTIVE: The purpose of this study was to examine the degree to which mechanical alterations in left ventricular papillary muscles of renal hypertensive-diabetic rat hearts correlate with functional measurements made on the same hearts in situ. METHODS: Female Wistar rats weighing 170-200 g were made hypertensive by placing a 0.24 mm clip on the left renal artery, and made diabetic 1 week later by a single intravenous injection of streptozotocin (60 mg/kg). Approximately 3-5 months later hemodynamic measurements including left ventricular pressure and dP/dtmax, arterial pressure and aortic flow were made on control and hypertensive-diabetic hearts in situ and correlated with mechanical measurements in left ventricular papillary muscles isolated from the same hearts. Body and tissue weights and biochemical and histological measurements were made at the time of sacrifice. RESULTS: Hypertensive-diabetic rats which survived to the time of study had decreased body weights, increased left ventricular weights and increased right ventricular weight to body weight and lung weight to body weight ratios. Those rats which died before the scheduled in-situ measurements had significantly more severe hypertension, greater left ventricular hypertrophy, increased right ventricular and lung weights, and more interstitial fibrosis than either surviving hypertensive-diabetics or controls. Rates of isometric tension development (normalized) and relaxation as well as shortening and relaxation velocities were significantly depressed in papillary muscles from hypertensive diabetic rat hearts despite unchanged developed tension and peak shortening. Time to peak tension and time to peak shortening were markedly prolonged. Mean aortic flow was maintained in the hypertensive-diabetic group despite significant depression of left ventricular dP/dtmax (normalized), peak aortic flow, peak aortic flow acceleration and heart rate. There was also significant depression of left ventricular-dP/dtmax. Ejection duration was markedly prolonged and correlated with both time to peak shortening in vitro and with stroke volume in vivo. CONCLUSIONS: Surviving hypertensive-diabetic rats were not in overt congestive heart failure; nevertheless, their hearts showed abnormal contractile performance which was qualitatively and quantitatively similar to that of left ventricular papillary muscles obtained from them. Depression of peak aortic flow, peak aortic flow acceleration and heart rate in the hypertensive-diabetic group was offset by increased ejection duration, resulting in normal mean aortic flow. The close correlation of ejection duration with time to peak shortening of the isolated papillary muscles suggests that it is a manifestation of an intrinsic change in the myocardium. To the extent that this prolongation is already maximized, further decreases in contractile speed would be expected eventually to cause depressed pump function and congestive heart failure. The possibility that this sequence of events occurred in the dying animals needs to be examined by evaluating in-vitro and in-vivo myocardial function at various stages of this disease model. PMID- 9217896 TI - Prevalence and prediction of silent ischaemia in diabetes mellitus: a population based study. AB - OBJECTIVES: The aim of the study was to estimate the prevalence of silent ischaemia in diabetic subjects in the population, to compare the prevalence of silent ischaemia in diabetics and non-diabetics and to attempt to predict the presence of silent ischaemia in diabetic subjects. METHODS: A random sample of 120 users of insulin and 120 users of oral hypoglycaemic agents aged 40-75 years living in the Danish municipality of Horsens were asked to participate in the study. Corresponding to the youngest half of the sample two non-diabetic controls were randomly selected from the Central Population Register. ST-depression of horizontal or descending character of at least 0.1 mV measured 80 ms after the J point on either exercise ECG or Holter ECG was considered indicative of myocardial ischaemia. Angina pectoris was considered present if the Rose questionnaire was positive, or chest pain was registered simultaneously with ECG evidence of ischaemia. Individuals with ischaemia, but without angina pectoris, were defined as persons with silent ischaemia. RESULTS: Seventy-four percent of the invited group were included. The observed prevalence of silent ischaemia in diabetics was 13.5% (95% CI = 8.5-19.8%). No association was found between silent ischaemia and gender (P = 0.83) or diabetes type (P = 0.67). In the group of diabetics who had controls, the prevalence was 11.4%, and among the controls the prevalence was 6.4% (OR = 1.87, one-sided P = 0.079). Systolic blood pressure was highly predictive of silent ischaemia in the diabetic subjects (P = 0.005). No predictive value could be shown for other variables. CONCLUSION: This is the first population-based study of silent ischaemia in diabetes. The prevalence of silent ischaemia in diabetic subjects was 13.5%. The frequency of silent ischaemia did not differ significantly between diabetics and non-diabetics. Systolic blood pressure was predictive of silent ischaemia in diabetes. PMID- 9217898 TI - Medical humanities in medical education. PMID- 9217899 TI - Medical education reform in South-East Asia: WHO perspectives. PMID- 9217897 TI - Mortality prediction in diabetic patients with myocardial infarction: experiences from the DIGAMI study. AB - OBJECTIVES: We analysed predictors of 1-year mortality following acute myocardial infarction in patients with diabetes mellitus by applying uni- and multivariate statistics on the DIGAMI cohort. BACKGROUND: Diabetic patients with acute myocardial infarction have a poor prognosis. This may depend on a poor metabolic control, a hypothesis that was tested in DIGAMI, a prospective randomised study. In this trial institution of immediate intensive insulin treatment reduced 1-year mortality by 30%. METHODS: We recruited 620 diabetic patients with acute myocardial infarction, 314 of whom served as controls, while the remaining 306 patients were treated with an acute insulin-glucose infusion followed by multidose subcutaneous insulin. RESULTS: Age, previous myocardial damage, duration of the diabetes and previous insulin therapy were significantly related to 1-year mortality, while conventional risk factors lacked independent prognostic weight. Female sex was not linked to mortality when controlling for the confounding effects of other predictors. One of the strongest predictors of a fatal outcome, in particular during the hospital phase, was blood glucose at hospital admission. Beta-blockade appeared to exert a striking, independent secondary-preventive effect. CONCLUSIONS: It seems that good metabolic control and not conventional risk factors is of major importance for diabetic patients sustaining acute myocardial infarction. Also treatment with beta-blockade seems to be of special importance in this category of patients. PMID- 9217900 TI - International co-operation for medical education and practice: a view from Thailand. AB - Medical practice and medical education are undergoing revolutionary changes all over the world. Even though the denominators of change are usually viewed at national and institutional levels, global trends also play an important role, and international co-operation is needed now, more than ever. Thailand with its rapid rate of change serves as an interesting example for review. PMID- 9217901 TI - A learning resources centre: its utilization by medical students. AB - Innovations in medical education have led to the increasing use of problem-based learning techniques, a committee system organization, and more time for independent study in many undergraduate programmes. There has been an increase in availability of alternative methods for presentation of information. To encourage self-directed learning, resources such as computers, videos and models, among others, should be readily available to students. The University of Calgary Faculty of Medicine has provided various resources contained in one area, called the Bacs Learning Resource Centre (BLRC). Since the maintenance and further development of such a centre requires significant resources, it is important to determine student utilization of the various components used in their learning. For those who are about to set up such a learning resource centre this information gives guidance on which materials are most useful. The utilization of the centre by 69 first year medical students was studied using questionnaires. The utilization during a specific course was determined by analysing the entries in the individual log books given to the students at the beginning of the Integrative Course. With the exception of one student, all those who responded to the questionnaire used the Centre, with 20% or less of their total study time being spent there. The BLRC was most used for the Musculoskeletal, Cardiovascular and Reticulo-Endothelial courses. All categories of resources were found to be useful, with the tape/slides least utilized. Utilization was most influenced by the quality of resources available and recommendations by peers. The development of a centre such as the BLRC, with a variety of resources concentrated in one area, suitable for individual or group study and accessible 24 hours a day, should be considered by all medical schools to enhance self-directed learning in medical students. PMID- 9217902 TI - Teaching colloquial Australian English to medical students from non-English speaking backgrounds. AB - Lack of fluency in the language of instruction can form a barrier to medical education. There has been an effort within Australian universities to teach English to students from non-English speaking backgrounds (NESB), but little systematic attention has been given to the teaching of informal or colloquial English. This paper provides evidence that colloquial language is a pervasive and important aspect of doctor-patient communication. It describes a teaching project for NESB medical students which aimed to introduce them to colloquial English, and to provide them with a contextual approach to learning this form of language. Forty-four first year medical students enrolled at the University of Adelaide were required to gather examples of colloquial language by interviewing a native English speaker. Ninety-four examples of colloquial sayings were recorded. These were compiled in the form of a handbook which served as a student resource. Student evaluation of this exercise was positive. The benefits of an interactive method of teaching local and setting-specific language are discussed, and the implications of this approach for clinical teaching and for medical practice are explored. PMID- 9217903 TI - Medical school entrance and career plans of Malaysian medical students. AB - This study investigates the reasons for entry to medicine and the career perspectives of phase III medical students of the Universiti Sains Malaysia (USM). The majority of the students were Malays from low socio-economic backgrounds who entered medical school after completing a 2-year matriculation course. An interest in medicine and helping people were the two main stated reasons for entry to medical school. A group of students wishing to work in private practice was identified. In comparison to the rest of the study body, students in the group were: not well prepared to enter medical school; dissatisfied with the course; and subject to family influences. A desire for monetary gain motivated their choice of medicine as a career. Overall, 13% of the students wished to change career because they were dissatisfied with their experience of medicine as undergraduates. The study did not find a significant difference in career intentions between female and male medical students. However, women were less likely to seek entrance into private practice or pursue formal postgraduate education. The choice of surgery as a career was confined to men. About 90% of the students had already decided on their future specialty. Four well-established specialties were their most popular choices. The gender of the students had no significant influences of the decision to continue into postgraduate education. The proportion of female students who wished to marry doctors was significantly higher than for male students. PMID- 9217904 TI - European medical schools and tobacco. AB - Following a survey in 19 European countries of the habits, attitudes and knowledge of medical students regarding tobacco, World Health Organisation European Office and the International Union against Tuberculosis and Lung Disease jointly circulated to the Deans of all European medical schools a summary of the results, including figures for mortality for smoking-related diseases in their countries and a brief questionnaire concerning faculty action on the tobacco problem. The response rate was just over 50%, higher in Northern Europe (66%) than in Southern (35%) or Eastern (38%). Only 8% of faculties had a specific teaching module on tobacco. In most it was either systematically (35%) or unsystematically (55%) integrated in other teaching. Teaching hospitals, teaching areas and faculty meetings were said to be smokefree by over 90%; figures were lower for other areas. Seventy-seven per cent of Deans intended to discuss our approach with their teaching staff; 72% gave the name of a staff member with a particular tobacco interest. PMID- 9217905 TI - Voluntary active euthanasia and doctor-assisted suicide: knowledge and attitudes of Dutch medical students. AB - The objective of the study was to gain insight into the knowledge of and attitudes towards voluntary active euthanasia and doctor-assisted suicide (EEDAS) of Dutch medical students, and to determine whether knowledge and attitudes change after a 1-day informative conference about EDAS. Data were collected by means of two self-administered questionnaires. Questionnaire 1 had to be completed before the start of the conference and questionnaire 2 after the conference. In both questionnaires, students were asked by means of two open ended questions to define euthanasia and doctor-assisted suicide. They were also asked to indicate which of eight statements met with the requirements for prudent practice. Finally, the students were asked to what extent they agreed or disagreed with each of seven statements about attitudes towards EDAS. To determine if a selection occurred among students who returned both questionnaires, their background characteristics, and knowledge and attitudes towards EDAS were compared with those who returned only the first questionnaire. Forty-seven students returned only the first questionnaire, while both questionnaires were returned by 137 students. No differences were found between students who returned both questionnaires and those who returned only the first questionnaire with regard to age, religion, knowledge of and attitudes towards EDAS. Students' knowledge of the definitions of EDAS and the requirements for prudent practice improved significantly. Students' reactions to the statements on attitudes towards EDAS showed that a large majority had a fairly positive attitude towards EDAS. There was no significant difference before and after the conference. Male students and students with a religion were more opposed to EDAS than female students and students without a religion. The fact that the students' knowledge of EDAS improved after a 1-day conference does not imply sufficient understanding of the issue. Because EDAS is allowed only under strict conditions in the Netherlands, medical students require special training. Only then will they be equipped to deal with requests for EDAS during their future careers. PMID- 9217906 TI - Death certification: production and evaluation of a training video. AB - The purpose of this study was to produce an effective training video on death certification suitable for use by medical students and postgraduates. A 15-minute video was commissioned from a video production unit and two authors (PA and CP) provided advice and support in the process of script writing and production. An evaluation by means of a randomized controlled trial took place among 185 first year medical students at the University of Leicester. The video was shown as an addition to the usual lecture on death certification. Performance in a test of knowledge, skill and motivation was recorded in each of the two groups. Students assigned to see the video scored slightly better overall in a test of knowledge and skill (difference in medians = 3, in a test marked out of 68, P = 0.046). The intervention group also gave a significantly higher priority to avoiding distress caused to relatives as a reason for certifying death accurately (60% vs. 35%, difference in proportions = 24%, P = 0.002). There was no evidence that enjoyment or views about the nature or content of the video had an impact on performance in the test. It is concluded that adding the video to the usual lecture had a limited effect on the overall knowledge and skills of undergraduate students but was highly effective in conveying the message that inaccurate death certification can cause distress to relatives. The randomized controlled trial is a practical and simple means of evaluating teaching methods for medical undergraduates. PMID- 9217907 TI - Competency-based learning in neonatology. AB - An educational model integrating structured teaching with clinical experience, or clerkship, has been designed to enable students to learn the core knowledge, skills and attitudes necessary to care for the newborn. The programme is run for fifth year medical students as part of four, 9-week periods in Obstetrics and Gynaecology each year. A SCORPIO teaching session is held in week 1 to introduce students to the core competencies in the subject areas of newborn examination, breastfeeding, resuscitation, respiratory distress and anthropometry. Groups of four students rotate through each topic, which is conducted by a neonatologist or registrar in training. Eight problem-based learning sessions are held during weeks 2-9. Several students assess a clinical problem, identify learning issues and meet colleagues and a facilitator to share their learning experiences and resolve the problem. The clinical experience, or clerkship, is based in the neonatal nursery where 2-3 students spend a week consolidating their clinical and procedural skills. A study group was assessed at the end of the programme by an Objective Structured Clinical Examination (OSCE) and a Multiple Choice Questionnaire (MCQ). A control group did the same assessment in week 1. All students were asked to rate the educational value of the three learning methods on a 5-point Likert scale. The study group (n = 20) achieved a mean composite mark of 66% (SD 10%). This was significantly higher (P < 0.001) than that of the control group (n = 18), mean 45% (SD 7%). All students (100%) rated the educational value of SCORPIO as high or very high, and the comparative rating for problem-based learning and clerkship was 65%, respectively. The programme was enthusiastically received by the students and resulted in mastery of a range of core competencies necessary for care of the newborn. PMID- 9217908 TI - American medical students in Israel: stress and coping. AB - Medical students studying abroad have to adapt to a new cultural environment in addition to the usual stresses of medical school. This study explored the perceived stress and coping ability of students of the New York State/American Programme, Sackler School of Medicine, Tel Aviv University, who study medicine in Israel but are expected to return to America to practice. Students were surveyed using the Ways of Coping Checklist (WCCL), Appraisal Dimension Scale (ADS) and two instruments specifically designed for the study. The results supported the view that students having difficulty adapting to their new cultural environment also have difficulty at medical school. This pattern is a negative spiral in which anxiety and depression impair cognitive performance, which leads to academic difficulties and emotional distress. Improvements in student social support and primary prevention were implemented as a result of the study. Limitations of the study are discussed. PMID- 9217909 TI - Dental and life science students: a comparison of approaches to study and course perceptions. AB - Approaches to study and course perceptions were investigated and compared in both junior and senior students on a vocational course (dentistry) and on a non vocational course (life sciences) using standard questionnaires. Junior dental students reported significantly greater tendencies towards both Deep and Surface Approaches than junior life science students. They also had significantly higher scores for Achieving Approach, but in senior students this trend was reversed. Final year dental students experienced greater workloads and less freedom in learning than life science students and they also reported less organized study methods. These course perceptions demonstrate similar trends to those reported by Ramsden (1991). The implications for the courses concerned and for curriculum development are considered. PMID- 9217910 TI - Research fellowships for medical students in Norway. PMID- 9217911 TI - Medical education in the Russian Federation. AB - The process of becoming a medical doctor in the Russian Federation is detailed in this paper. There has been a decline in the number of students entering the medical profession, as well as a marked decrease in the faculty members at the medical institutes since perestroika. This is secondary to a marked decrease in financial support as well as falling morale. PMID- 9217913 TI - Generally applicable competences. PMID- 9217914 TI - Long-term effect of minor surgery training workshops for general practice registrars. PMID- 9217912 TI - Junior doctors and HIV--are they aware? PMID- 9217915 TI - The hemodynamic manifestation of normal myocardial relaxation. A framework for experimental and clinical evaluation. AB - Myocardial relaxation clinically manifests itself as left ventricular pressure (LVP) fall. The transition from contraction to relaxation is the precise moment at which 81-84% of peak isometric force has developed or the equivalent timing early during ejection. Defining the completion of relaxation and distinguishing relaxation from diastole appears merely semantic. Diastole is not a passive phase of the cardiac cycle. During diastole mechanical left ventricular properties still change due to incomplete relaxation, due to creep and stress relaxation, and due to autoregulation by preload and by nitric oxide. Description of timing and rate of LVP fall may provide useful information on underlying cardiac diseases such as ischaemia and hypertrophy. This information will however only be reliable if systolic cardiac function and systolic load are normal, and in the absence of a significant degree of nonuniformity, such as induced by conduction disturbances or by regional myocardial ischemia. The various effects of load and of nonuniformity on myocardial relaxation in the normal heart are reviewed. Coupling of timing and rate of LVP fall are explained in terms of cross-bridge mechanics. Specific effects of systolic pressure on LVP fall and their relation to systolic cardiac function are emphasized. These data constitute a conceptual framework for the analysis of myocardial relaxation in cardiovascular research and in the cardiac patient. Comparison of clinical and experimental data during manipulation of afterload should lead to an improved understanding of clinical relaxation disturbances and to a therapeutic approach, which is relevant from the physiopathological point of view. LVP fall may provide useful and quantitative information on systolic LV function if measurements are performed under different conditions of systolic load. This information is similar to systolic pressure volume relations, but can be performed with the sole use of a micromanometer in the LV cavity. PMID- 9217917 TI - Different secretion profiles of atrial and brain natriuretic peptides after acute volume loading in patients with ischemic heart disease. AB - In order to clarify the different secretion profiles of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP) in response to acute hemodynamic change by volume expansion, we measured plasma ANP and BNP levels after intravenous isotonic saline infusion for 3 min at a rate of 3 ml/kg body weight/min in 15 patients with ischemic heart disease. Plasma ANP and BNP levels before the volume loading were 30.7 +/- 16.7 and 19.4 +/- 24.6 pg/ml, respectively. Five and 10 minutes after infusion, plasma ANP levels rose significantly to 43.5 +/- 20.7 and to 46.0 +/- 22.5 pg/ml, respectively (p < 0.01), and plasma BNP levels rose significantly to 27.3 +/- 30.8 and 24.8 +/- 23.2 pg/ ml, respectively (p < 0.01). The BNP/ANP ratio was not affected by volume loading. The maximum increments of plasma ANP level correlated significantly with those of the mean pulmonary capillary wedge pressure (mPCWP, r = 0.78, p < 0.01) or left ventricular end-diastolic pressure (LVEDP, r = 0.86, p < 0.01). However, there were no significant correlations between the maximum increments of plasma BNP levels and those of mPCWP or LVEDP. Plasma ANP level can be a useful parameter for atrial pressure even if the hemodynamic state change rapidly. However, in an early phase of ventricular overload BNP secretion is not increased sufficiently despite the raised LVEDP, and plasma BNP level may not always reflect ventricular hemodynamics. PMID- 9217916 TI - The SF-36 questionnaire: a convenient way to assess quality of life in angina pectoris patients. AB - Over 4400 stable angina pectoris patients were followed for six months, after the addition of nitroglycerin patches to their usual treatment. Qol items were assessed by means of the SF-36 questionnaire and some clinical parameters were followed as well: during the study, the frequency of attacks decreased regularly and significantly (from 3.49/ week to 0.99/week), as did the amount of sublingual nitroglycerin tablets (from 2.65/day to 0.50/day). All 8 items of the SF-36 questionnaire improved significantly. Moreover, there was a modest, but significant correlation between the decrease in attacks and rescue-medication and the improvement in the Qol. The SF-36 questionnaire appears to be an adequate tool to follow the changes in Qol in stable angina pectoris patients over time. PMID- 9217918 TI - Caffeine and cardiac arrhythmias. An experimental study in dogs with review of literature. AB - In spite of widespread belief among clinicians that caffeinated drinks are linked with palpitations, tachycardia and dysrhythmia there is paucity of documentary evidence. We investigated the arrhythmogenic activity of caffeine in canine model. The alkaloid was given i.v. to perform 51 experiments in 13 anesthetized dogs in three different doses. The Low dose generated significant Sinus (S) bradycardia (70%; p < 0.006, S. arrhythmia (70%; p < 0.02), S. arrest (50%; p < 0.04), Atrial (A) ectopics (40%; p < 0.016), Wandering of pacemaker (WPM) (50%; p < 0.04), and Ventricular premature contractions (VPC--unifocal 40%; p < 0.05) as compared with control ECGs. Medium dose induced significant S. arrhythmia (62%; p < 0.001), A. ectopics (25%; p < 0.01), A. tachycardia (25%; p < 0.01), WPM (25%; p < 0.01), VPCs--unifocal (50%; p < 0.002), multifocal (25%; p < 0.01), couplets (25%; p < 0.01) and interpolated (25%; p < 0.01). High dose of caffeine revealed significant S. arrhythmia (56%; p < 0.002), A. ectopics (44%; p < 0.005), A. tachycardia (32%; p < 0.01), WPM (32%; p < 0.01) and VPCs--unifocal (64%; p < 0.001), multifocal (32%, p < 0.01), couplets (32%, p < 0.01), interpolated (32%; p < 0.01) and Ventricular tachycardia (VT) (20%; p < 0.01). A. flutter and fibrillation each were observed in two experiments only. In conclusion, these data indicate a dose dependent arrhythmogenecity of caffeine. Small dose, mostly, generated, benign arrhythmias due to vagal stimulation. More severe arrhythmias like VT, multifocal VPC. A. flutter and A. fibrillation were generated with higher dose of caffeine. Mechanism remains uncertain as caffeine has multiple actions. Further studies in human beings with normal and compromised myocardium may elucidate arrhythmogenic effects of caffeine. PMID- 9217919 TI - Prolongation of the QTc interval during alcohol withdrawal syndrome. AB - BACKGROUND: Cardiac arrhythmias might explain cases of sudden death in alcoholics during ethanol abstinence. AIM OF STUDY: To evaluate QT interval (and its outcome) in patients with alcohol withdrawal syndrome. PATIENTS AND METHODS: Sixty-two patients (52 male and 10 female, mean age 43.7 years, range 18-71 years), admitted to the hospital as a result of alcohol abstinence syndrome (tremulousness in 7 cases, agitation in 12 cases, delirium tremens in 11 cases, and seizures in 32) were studied. The QT interval was measured on 12 lead ECGs performed on admission in all cases. QTc was obtained using Bazett's formula. In 27 patients a second ECG was performed during hospital stay. Results. Mean QTc interval on admission was 439 +/- 32 ms (range 350-525 ms); 29 patients (46.8%) showed a prolonged (> 440 ms) QTc interval. No significant differences were observed between patients with normal and high QTc values as regards to age, sex, type of withdrawal syndrome, duration of abstinence, liver function tests, serum electrolytes or blood cell counts. In cases where two ECG recordings could be evaluated (n = 27), the mean QTc interval was significantly shorter in the latter than in the former (417 +/- 26 ms versus 447 +/- 30 ms, respectively, p < 0.001). Eight patients found to have prolonged QTc on admission had a second ECG performed on them after complete recovery from withdrawal symptoms. In all these cases the QTc interval eventually became normal. CONCLUSION: The QTc interval is frequently prolonged during alcohol withdrawal syndrome and tends to become normal over time, along with remission of abstinence symptoms. PMID- 9217920 TI - Complete left bundle branch block with left QRS axis deviation: defining its clinical importance. AB - Aim of this study was to elucidate the diagnostic significance of left axis deviation (LAD) in patients (pts) with chronic (> 6 months) left bundle branch block (LBBB). We retrospectively studied 2094 consecutive pts who underwent cardiac catheterization. All pts had left heart catheterization and coronary angiography, whereas right heart catheterization or endomyocardial biopsy were performed on indication. Our study group consisted of 43 pts with LBBB (29 men, 14 women, mean age 60.3 +/- 7.9 years). Pts with acute myocardial infarction or prior high degree AV-block were excluded. Initial evaluation included history, physical examination, chest X-ray, serial ECGs, 2D-echo and Doppler studies. ECG criteria for LBBB were a QRS duration of > 0.12 secs, a predominantly negative QRS deflection in V1 and a widened R-wave in V6. LAD was considered present when the mean frontal QRS axis was between -30 degrees and -90 degrees. The mean frontal QRS axis was considered normal if it was between -29 degrees and +90 degrees. Twenty-nine pts had normal axis and 14 had LAD. According to angiographic data, among coronary disease pts, 12 (31.48%) had normal axis and 4 (28.57%) LAD (p = 0.041). Among mitral valve disease pts, 3 (10.35%) had normal axis and none LAD. Among pts with aortic valve disease, I (3.45%) had normal axis and 8 (57.15%) LAD (p = 0.0001). Among pts with dilated cardiomyopathy, 2 (6.9%) had normal axis and 1 (7.14%) LAD. Among pts with no organic heart disease, 11 (37.93%) had normal axis and 1 (7.14%) LAD (p = 0.035). The presence of LAD had a 41.9% sensitivity and a 91.6% specificity for the presence of organic heart disease. These findings point towards a statistically significant difference in the presence of organic heart disease in LBBB pts with LAD compared to normals. Aortic valve disease in LBBB pts seems to be frequently accompanied by LAD. PMID- 9217921 TI - Impact of hemodialysis on comprehensive ventricular repolarization. AB - Cardiac arrhythmia are one of the most important problems in haemodialysis patients. An important cause of the arrhythmias is inhomogenous myocardial repolarization. In this study, the ventricular repolarization parameters (QT, QTc, JT and JTc) and dispersions (d) of the parameters (QT-d, QTc-d, JT-d and JTc d) were evaluated. Also were recorded the right-sided leads (RV3-6) and posterior leads (V7-9) in addition to the standard 12 lead ECG to assess comprehensive ventricular repolarization. The leads were divided in three groups: Group A (Standard ECG leads), Group B (Right-sided leads) and Group C (All of the leads). Among the above mentioned parameters, only JT and JTc intervals decreased significantly in all groups. There was no significant difference between the groups in evaluation of the parameters. It was concluded that in assessment of ventricular repolarization, the most important ECG intervals may be JT and JTc intervals, and the standard 12 lead ECG record is sufficient in evaluation of ventricular repolarization in hemodialysis patients. PMID- 9217922 TI - Advances in understanding the pharmacological properties of antisense oligonucleotides. PMID- 9217923 TI - Targeted tumor cytotoxicity mediated by intracellular single-chain anti-oncogene antibodies. PMID- 9217924 TI - In vivo gene therapy with adeno-associated virus vectors for cystic fibrosis. PMID- 9217925 TI - Engineering herpes simplex virus vectors for human gene therapy. PMID- 9217926 TI - Human adenovirus vectors for gene transfer into mammalian cells. PMID- 9217927 TI - Anti-oncogene ribozymes for cancer gene therapy. PMID- 9217928 TI - Cytokine gene transduction in the immunotherapy of cancer. PMID- 9217929 TI - Gene therapy approaches to enhance antitumor immunity. PMID- 9217930 TI - Modified steroid receptors and steroid-inducible promoters as genetic switches for gene therapy. PMID- 9217931 TI - Strategies for approaching retinoblastoma tumor suppressor gene therapy. PMID- 9217933 TI - Antisense inhibition of virus infections. PMID- 9217932 TI - Immunoliposomes for cancer treatment. PMID- 9217934 TI - Two products join ranks of smoking cessation treatments. PMID- 9217935 TI - Study suggests antioxidants slow decline in Alzheimer's disease. PMID- 9217936 TI - Practice guidelines cover management of Alzheimer's disease. PMID- 9217937 TI - Thalidomide heals HIV-related oral ulcers, but caution is required. PMID- 9217938 TI - Program on medication misuse airs on PBS. PMID- 9217939 TI - The pharmacist as a health education coordinator. PMID- 9217940 TI - Making software work for pharmacy. PMID- 9217941 TI - Building the clinical workstation: software for the health-system pharmacist. PMID- 9217942 TI - Building clinical decision-support systems to profile physician practice and develop guidelines in a staff-model HMO. PMID- 9217943 TI - Clinical information system in a mixed-model HMO. PMID- 9217944 TI - Satisfaction among residents in ASHP-accredited pharmacy residency programs. AB - The level of work satisfaction among pharmacists in ASHP-accredited residencies was studied. In March 1996 a questionnaire designed to measure residency satisfaction was mailed to 697 individuals in ASHP-accredited pharmacy practice and specialty practice residencies. Subjects responded to 16 statements relating to intrinsic and extrinsic determinants of work satisfaction on a scale of 1 to 5, where 1 = strongly disagree and 5 = strongly agree. Questionnaires were returned by 413 (59%) of the residents. The respondents were predominantly women (76%), and most (86%) had at least a Pharm. D. degree. Hospitals were the primary work setting (88%). Of the 413 residents, 305 were in pharmacy practice residencies and 108 were in specialized residencies. None of the mean scores indicated disagreement (scores < 3) with the positively worded statements or agreement (scores > 3) with the negatively worded statements. The median and mode were equal to 2 (disagree) for the three negatively worded items and 4 (agree) for all but three positively worded items. Only 8% of the residents indicated that they would not accept the residency again if given the chance. Specialized residents tended to rate positively worded statements higher and negatively worded statements lower than pharmacy practice residents. Female residents indicated greater satisfaction than male residents. Pay and benefits were rated slightly better than neutral. Pharmacy residents appeared generally satisfied with their residencies. Specialized pharmacy residents were more satisfied than pharmacy practice residents, and women were more satisfied than men. PMID- 9217945 TI - Quality measurement for health systems: accreditation and report cards. AB - Efforts to measure the quality of care provided by health systems, including the accreditation processes used by the National Committee for Quality Assurance (NCQA) and the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) and three report card systems-NCQA's Health Plan Employer Data and Information Set (HEDIS), JCAHO's requirements for performance data (Oryx), and a consumer-oriented system of performance measures created by the Foundation for Accountability (FACCT)-are described. NCQA reviews and accredits all types of health maintenance organizations, as well as point-of-service health plans and certain physician-hospital organizations. NCQA accreditation survey teams consist primarily of physicians. The fee for the process-oriented survey depends on the size and complexity of the plan. HEDIS was created to standardize the way in which health plans calculate and report information about their performance, thereby enabling comparisons among plans. NCQA is including more outcome measures as HEDIS is revised. Several HEDIS measures relate to medication use. In the future, health plans will be required to submit HEDIS data as part of the NCQA survey process. JCAHO's accreditation program for health care networks began in 1994. Survey teams and fees depend on the size and complexity of the network. A survey covers the central office, all hospitals, and selected components (excluding those that are accredited by other bodies recognized by JCAHO), and practitioners' offices. NCQA and JCAHO have no pharmacy-specific accreditation standards, but pharmacy information may be requested by the surveyors. In 1997 JCAHO began phasing in requirements for performance data as part of its accreditation process; networks must begin collecting data for at least 10 measures from five approved systems. FACCT performance measures are based on patients' health care experiences and outcomes; FACCT hopes these measures will be adopted by NCQA, JCAHO, and others. As health care purchasers and consumers increasingly demand evidence of quality, pharmacists will have ample opportunity to improve health outcomes. PMID- 9217946 TI - Restarting trimethoprim-sulfamethoxazole after a hypersensitivity reaction in patients infected with the human immunodeficiency virus. PMID- 9217947 TI - Probable interaction of warfarin and acarbose. PMID- 9217948 TI - Repeated courses of steroids in preterm membrane rupture do not increase the risk of histologic chorioamnionitis. AB - Antenatal administration of steroids (betamethasone 12 mg i.m. twice q 24 hr) enhances fetal lung maturation and reduces neonatal morbidity in preterm prelabor rupture of membranes (PROM). However, the risks of repeated administration of steroids 7 days after the initial course are unknown. We evaluated the prevalence of histologic evidence of chorioamnionitis in patients receiving single versus multiple courses of steroids for fetal lung maturation. We performed a retrospective analysis of consecutive cases of preterm PROM at < 32 weeks' gestation prospectively collected between July 1988 and March 1994. Obstetric and clinical information were obtained for women who did not receive steroids for fetal lung maturity (n = 55), those who received a single course (n = 47), and those with > or = 2 courses of steroids (n = 89). Placental pathology examination was performed after delivery, and histologic evidence of acute placental inflammation was determined and scored semiquantitatively on a scale of 0-4, as previously described. Potential confounding variables considered were: presence of oligohydramnios (vertical pocket of amniotic fluid < or = 1 cm at ultrasound), onset of labor prior to delivery, gestational age at delivery, and mode of delivery. The three groups were comparable for gestational age at membrane rupture and at delivery, rate of oligohydramnios, labor prior to delivery, and mode of delivery. Administration of multiple courses of steroids was associated with a decrease in the rate of clinical chorioamnionitis (p < 0.0001) and severity of histologic acute placental inflammation (mean +/- SD total score of acute inflammation: 10.7 +/- 7.2 vs. 7.3 +/- 6.0 vs. 6.9 +/- 6.0, p = 0.005) compared with the groups receiving no steroids or administration of a single course of steroids. In preterm PROM at < 32 weeks, repeated administration of courses of steroids is not associated with an increase in the prevalence of clinical or histologic evidence of infectious outcome. These findings may reflect a greater likelihood for noninfected patients to remain quiescent and thus receive repeated courses of steroids. PMID- 9217949 TI - Intrapartum sonography of the lower uterine segment in patients with breech presenting fetuses. AB - Uterine leiomyomas have been associated with an increased incidence of fetal malpresentation. Intrapartum sonographic examination, currently advocated in the assessment of patients with a breech presenting fetus includes assessment of flexion of the fetal head, sonographic estimated fetal weight, and the precise type of breech presentation. Data obtained during this evaluation is utilized in deciding the preferred mode of delivery. We report two cases in which intrapartum sonographic assessment of a women with breech-presenting fetuses at 39 and 40 weeks of gestation depicted large lower uterine segment leiomyomas, which were considered an indication for cesarean. The association between uterine leiomyomas and fetal malpresentation, and our cases support consideration of sonographic scanning of the lower uterine segment in patients being evaluated for vaginal delivery with a breech-presenting fetus. PMID- 9217950 TI - Isolated persistent fetal bradycardia in complete A-V block: a conservative approach is appropriate. A case report and a review of the literature. AB - Persistent fetal bradycardia is rarely encountered during pregnancy. When it is associated with a complete atrio-ventricular (A-V) block, it may prove dangerous to the fetus or newborn. The prenatal diagnosis is vital because it necessitates close follow-up during pregnancy to detect fetal compromise and proper preparation for delivery. We describe a woman who was found to be suffering from systemic lupus erythematosus during pregnancy. The fetus was diagnosed as having persistent fetal bradycardia due to complete A-V block at 28 weeks of gestation and was delivered at term with conservative management. The problems entailed in managing pregnancy and delivery of such fetuses are discussed. PMID- 9217951 TI - Spontaneous rectus sheath hematoma during pregnancy mimicking abruptio placenta. AB - Sudden disruption of a deep epigastric vessel may result in an abdominal wall hematoma, which, depending upon its location and size, can produce symptoms and clinical findings compatible with a variety of acute intra-abdominal conditions. The literature has noted a predominance of pregnant patients among those affected with this malady. Such hematomas are infrequently encountered and early accurate diagnosis could prevent surgical intervention. Unfortunately, the clinical manifestations of rectus muscle hematoma are sometimes so dramatic that laparotomy is performed under the belief that intra-abdominal pathology is present. We present a case of a suspected abruptio placenta misdiagnosed by clinical and ultrasound examination that was subsequently discovered to be a rectus sheath hematoma at the time of surgery. PMID- 9217953 TI - Trauma and pregnancy. AB - Trauma and/or accidental injury complicates 6-7% of all pregnancies. The management protocols for trauma in pregnancy are based largely on case reports and small series. The purposes of this study were to: describe the demographics of pregnant trauma patients at a tertiary care center and a large community hospital; identify variables predictive of fetal outcome including an examination of Kleihauer-Betke and nonstress testing; and recommend an evaluation and management protocol after trauma based on empirical data rather than anecdotal reports. Data from pregnancies complicated by trauma from July 1987 through October 1993 were retrospectively reviewed. Statistical analysis included Chi square and Kruskall-Wallis testing. There were 476 medical records available for review. Of the trauma cases, 54.6% were motor vehicle accidents, 22.3% were domestic abuse and assaults, 21.8% were associated with falls, and 1.3% were secondary to burns, puncture wounds, or animal bites. Mean maternal age was 24 years, 49.9% were Caucasian, and 43% were primigravid. Mean gestational age at occurrence of trauma was 25.9 weeks and mean gestational age of delivery was 37.9 weeks. Domestic abuse occurred most frequently before 18 weeks, falls between 20 30 weeks' gestation, and motor vehicle accidents occurred with equal frequency throughout gestation. Uterine contractions occurred in 39.8% of patients and as often as every 1 to 5 min in 18% of patients. Preterm labor occurred in 11.4%, preterm delivery in 25%, and abruptions in 1.58% of the trauma population. Fetal heart rate monitoring was abnormal in 3% of cases. Twenty-seven perinatal deaths were noted and in 14 pregnancies the deaths were related to trauma. Eight of these perinatal deaths were associated with motor vehicle accidents, four with domestic violence, and two with falls. The only preventable perinatal deaths were a twin pregnancy transferred with nonreassuring fetal heart tones. Early warning symptoms of vaginal bleeding, uterine contractions, and/or abdominal and/or uterine tenderness were not predictive of either preterm delivery or adverse pregnancy outcome, sensitivity 52%, specificity 48%. Abnormal monitoring and positive Kleihauer-Betke tests were also not predictive of adverse pregnancy outcome. However, there were no adverse outcomes directly related to trauma when monitoring was normal and early warning symptoms were absent (negative predictive value 100%). Two hundred eighty-nine Kleihauer-Betke tests were performed and only one affected management. Repetitive monitoring over several days did not uncover any patients whose heart rate tracings evolved from normal to abnormal monitoring. Given our findings that prolonged monitoring was not helpful in management of pregnant trauma patients, we support the recommendation that initial external fetal monitoring be performed for 4 hr, and, if reassuring, the patient may be sent home with precautions. We also recommend an Rh-immunoglobulin work-up for all Rh-negative pregnant trauma patients, but do not recommend Kleihauer-Betke testing for Rh-positive women. Given the frequency with which trauma affects pregnancy and the difficulty encountered with identifying variables predictive of pregnancy outcome, there may be great benefits of incorporating trauma prevention into routine prenatal care. PMID- 9217952 TI - Amnioinfusion for prevention of pulmonary hypoplasia in second-trimester rupture of membranes. AB - We conducted a study to evaluate the feasibility and benefits of transabdominal amnioinfusion in preterm premature rupture of membranes with persistent oligohydramnios for the prevention of pulmonary hypoplasia. To this purpose, we designed a cohort study in which the pregnancy outcome of women with rupture of membranes at < or = 25 weeks and persistent (> or = 4 days) oligohydramnios managed with serial amnioinfusions (n = 18) was compared with that of a historic cohort group (controls) with similar characteristics but managed expectantly (n = 16). Pulmonary hypoplasia was diagnosed at birth in the presence of strict radiological and pathological criteria. No amnioinfusion-related complications occurred. The prevalence of pulmonary hypoplasia was significantly lower among the amnioinfused cases compared with the controls (46% [6 of 13] vs 86% [12 of 14], odds ratio [OR] = 0.4, 95% confidence interval [CI] 0.2-0.9), despite a lower gestational age at rupture of membranes in the treated group. Within the group undergoing amnioinfusions, those in which the infused solution was rapidly lost had a higher rate of pulmonary hypoplasia compared with those in which amnioinfusion alleviated oligohydramnios for > 48 hours (considered successful) (0 of 4 vs. 6 of 9, OR = 2.3, 95% CI 1-5.5). Cases of successful amnioinfusion had a longer interval between membrane rupture and appearance of oligohydramnios than those in which the procedure failed to correct oligohydramnios, even though both groups had similar gestational age at appearance of oligohydramnios. This suggests that the rate of loss of amniotic fluid after membrane rupture may predict the rate of loss of the infused solution, and therefore identify a subset of patients who may benefit from the procedure. PMID- 9217954 TI - Fetal breathing movements within 24 hours of delivery in prematurity are related to histologic and clinical evidence of amnionitis. AB - Our objective was to determine the association of fetal breathing movements (FBM) within 24 hr of delivery, with the clinical diagnosis of chorioamnionitis and histologic evidence of severe acute amnionitis and umbilical-chorionic vasculitis in women delivering at < 32 weeks' gestation. We performed a cohort study of patients with singleton gestations delivered at < 32 weeks' following preterm labor with intact membranes and sonographically assessed biophysical profile within 24 hr of delivery (n = 111). Patients with FBM were compared with those without FBM, with regard to prevalence of clinical chorioamnionitis (CA) and histologic diagnosis of acute amnionitis and umbilical vasculitis. Maternal and neonatal charts were reviewed and the diagnosis of clinical CA made by previously established criteria. Histologic presence and extent of acute intrauterine inflammation was assessed and scored by a single pathologist blinded to clinical information. Results are presented as chi 2 values and odds ratios with 95% confidence intervals. Of the patients included in the study, FBM were absent in 56 and present in 55. The prevalence of CA was 13% (15 of 111), severe acute amnionitis 34% (38 of 111), and severe umbilical vasculitis 23% (26 of 111). Severe umbilical vasculitis was significantly less frequent in cases with FBM as compared to cases without FBM (15% [8 of 55] vs. 32% [18 of 56], p = 0.049). However, the difference in rate of CA (22% [12 of 55] vs. 34% [19 of 56], p = 0.22) and histologic severe amnionitis (29% [16 of 55] vs. 39% [22 of 56], p = 0.4) between cases with and without FBM was not significant. In the presence of preterm labor with intact membranes, absence of FBM had sensitivities of 73 and 72%, and specificities of 54 and 56% in the prediction of CA and histologic evidence of umbilical vasculitis, respectively. We conclude that absence of FBM is associated with histologic evidence of fetal inflammation in intrauterine infection in patients with preterm labor and intact membranes delivering at < 32 weeks. However, the low positive predictive value of absent FBM in predicting fetal inflammation in intrauterine infection should discourage the guidance of clinical management in patients < 32 weeks' gestation with preterm labor and intact membranes. PMID- 9217956 TI - Laerdal infant resuscitators are unreliable as free-flow oxygen delivery devices. AB - Laerdal Infant Resuscitators (Laerdal Medical Co., NY) are commonly used as free flow oxygen delivery devices during neonatal resuscitation in situations where oxygen but not mechanical ventilation is desired. This study evaluates the performance of these resuscitators as free-flow oxygen devices. Efficiency was measured by comparing oxygen flow entering the resuscitator to oxygen flow delivered by the resuscitator. Clinical impact was assessed by measuring simulated patient fiO2. Three randomly selected resuscitator bags were tested for oxygen delivery efficiency by comparing oxygen inflow to outflow over an inflow range of 1 to 15 liters per minute (lpm). Measured outflow was 18-24% of inflow, demonstrating that as much as 82% of the oxygen flow escapes via the safety air inlet, safety blow-off valves, and other leaks. With the patient valve assembly removed, efficiency improved to 53-59%. Simulated fiO2 ranged from 0.23 to 0.68 at 5 lpm oxygen flow. We conclude that use of the Laerdal Infant Resuscitator for the delivery of free-flow oxygen, even with the valve assembly removed, generates highly variable patient fiO2. The use of self-inflating bags for delivery of oxygen without manual ventilation should be reconsidered. PMID- 9217955 TI - Attitudes toward health-care, HIV infection, and perinatal transmission interventions in a cohort of inner-city, pregnant women. AB - The objective of this article is to explore attitudes of an inner-city, pregnant cohort about general and HIV-related prenatal care. Responses to an interview at initial prenatal care enrollment were compared using Chi-square and Fisher's exact tests. Of 75 women, drug users (51%) were more likely to say that they would defer initiating prenatal care (P = 0.03) and to minimize the risk of drug or alcohol use to the fetus (P = 0.04). Most (85%) viewed pregnancy as inappropriate for HIV infected women and primarily drug users (P = 0.06) would abort if HIV infected. Over half thought HIV transmission occurred most times or always. Only 20% had heard of a drug to reduce this risk, but 95% would take such a therapy. These inner-city, pregnant women disapproved of pregnancy if HIV infected and thought the risk of transmission was high. They knew little of how to reduce this risk but nearly all would accept a drug to prevent transmission. PMID- 9217957 TI - Laryngeal obstruction caused by lingual thyroglossal duct cyst presenting at birth. PMID- 9217958 TI - Association of parvovirus infection with isolated fetal effusions. AB - The association of parvovirus B19 infection and hydrops fetalis is well known. However, the association of parvovirus and fetal pleural or pericardial effusions has not been reported. We present five cases of isolated pleural or pericardial effusion with documented maternal parvovirus infection in four of these pregnancies. In the absence of structural or karyotypic abnormalities, spontaneous resolution of the effusion portends for a successful pregnancy outcome. PMID- 9217959 TI - Oligohydramnios and the appropriately grown fetus. AB - The perinatal implications of oligohydramnios prior to 37 weeks of gestation, in the absence of intrauterine growth restriction (IUGR), rupture of membranes or fetal anomalies, are unknown. We compared the outcomes of 65 women with oligohydramnios (amniotic fluid index ([AFI] < or = 8 cm) by sonography to those of a control group matched by sonogram indication. Study patients were between 17 and 37 weeks of gestation, with appropriately grown fetuses on index sonogram and no other detected explanation for amniotic fluid abnormalities. Patients were managed expectantly with fetal testing and follow-up sonograms for fetal growth. Delivery was not recommended solely for oligohydramnios until 37 weeks of gestation. Patients with isolated oligohydramnios prior to 37 weeks of gestation, when compared to a control group with normal amniotic fluid volume, had a significantly higher incidence of premature delivery (odds ratio [OR] 3.23, 95% confidence interval [CI] 1.4-7.3) but did not appear to be at increased risk of IUGR, intrauterine death, or birth asphyxia. PMID- 9217960 TI - Ultrastructure of human colostral cells. AB - The ultrastructural architecture of colostral cells of mothers of pre- and full term infants is described. The polymorphonuclears were engaged in vivid phagocytosis of fat droplets. Similar findings were observed on the macrophages. The lymphocytes appeared normal in size and ultrastructure. A small number of eosinophils and basophils were also detected. The number of colostral cells was higher in the colostrum of mothers of preterm newborns. The number of the cells in the colostrum in mothers of both groups decreased with advancement of lactation. PMID- 9217961 TI - Dubowitz assessment of gestational age and agreement with prenatal methods. AB - We compared assessment of gestational age by Dubowitz score with ultrasonic measurement of the biparietal diameter (BPD), and then evaluated how infants were classified by these methods as small-for-gestational age (SGA), and as pre- or post-term births. BPD gestational age was assessed at week 17 to 20 of pregnancy while the Dubowitz scoring was done at birth. "Limits of agreement" between methods and kappa values were calculated and used to evaluate agreement. Among 839 included infants, there was moderate agreement between Dubowitz score and BPD (limits of agreement; -2.3; +2.1 weeks; weighted kappa: 0.46) in the assessment of gestational age. Agreement between Dubowitz score and BPD in the classification of SGA (kappa: 0.75, 95% confidence interval [CI]: 0.69-0.81) and preterm infants (kappa: 0.68, 95% CI: 0.56-0.80) was good, whereas agreement on infants born post-term was no better than chance (kappa: 0.14, 95% CI: -0.02( )+0.30). We conclude that despite moderate agreement between Dubowitz score and BPD in the assessment of gestational age, agreement in the classification of low birth-weight infants as SGA and as premature births was good. PMID- 9217962 TI - Cellular immune recognition and the biological role of major transplantation antigens. AB - The experiments leading to the discovery of MHC-restricted T-cell killing of virus-infected cells are described. The implications of MHC- and HLA-restricted T cell killing for the development of new vaccines and for the understanding of autoimmune disease are discussed. PMID- 9217963 TI - Genes of human ATP synthase: their roles in physiology and aging. AB - The reaction of ATP synthase (F0F1) is the final step in oxidative phosphorylation (OXPHOS). Although OXPHOS has been studied extensively in bacteria, no tissue-specific functions nor bioenergetic disease, such as mitochondrial encephalomyopathy and aging occur in these organisms. Recent developments of the Human Genome Project will become an important factor in the study of mammalian bioenergetics. To elucidate the physiological roles of human F0F1, genes encoding the subunits of F0F1 were sequenced, and their expression in human cells was analyzed. The following results were obtained: A. The roles of the residues in F0F1 are not only to transform the energy of the electrochemical potential (delta mu H+) across the membrane, but also to respond rapidly to the changes in the energy demand by regulating the intramolecular rotation of F0F1 with the delta mu H+ and the inhibitors of the ATPase. B. The roles of the control regions of the F0F1 genes, are to coordinate both mitochondrial DNA (mtDNA) and nuclear DNA (nDNA) depending on the energy demand of the cells, especially in muscle, C. The cause of the age-dependent decline of ATP synthesis has been attributed to the accumulation of mutations in mtDNA. However, the involvement of nDNA in the decline is also important because of telomere shortening in somatic cells, and age-dependent mtDNA expression analyzed with rho degree cells (cells without mtDNA). PMID- 9217964 TI - Nucleotide transport through the cystic fibrosis transmembrane conductance regulator. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) is a member of the superfamily of ATP-binding cassette (ABC) transporters, also known as traffic ATPases, which are implicated in the movement of various substrates. Recent studies indicate that CFTR and other closely related ABC transporters are also implicated in the movement of cellular ATP. This is the subject of current controversy. Therefore, evidence for the movement of cellular nucleotides by expression of CFTR and related molecules, as well as the potential significance of ATP-permeable channels in cell physiology, are reviewed in this study. The hypothesis is thus forwarded for the improper delivery of cellular ATP to the extracellular milieu by a dysfunctional CFTR, to be a relevant factor in the onset of cystic fibrosis. PMID- 9217965 TI - Significance of the beta-subunit in the biogenesis of Na+/K(+)-ATPase. AB - This review summarizes our experiments on the significance of the beta-subunit in the functional expression of Na+/K(+)-ATPase. The beta-subunit acts like a receptor for the alpha-subunit in the biogenesis of Na+/K(+)-ATPase and facilitates the correct folding of the alpha-subunit in the membrane. The alpha subunit synthesized in the absence of the beta-subunit is subjected to rapid degradation in the endoplasmic reticulum. Several assembly sites are assigned in the sequence of the beta-subunit from the cytoplasmic NH2-terminal domain to the extracellular COOH-terminus: the NH2-terminal region of the extracellular domain, the conservative proline in the third disulfide loop, the hydrophobic amino acid residues near the COOH-terminus and the cysteine residues forming the second and the third disulfide bridges. Upon assembly, the beta-subunit confers a resistance to trypsin on the alpha-subunit. The conformations induced in the alpha-subunit of Na+/K(+)-ATPase by Na+/K(+)- and H+/K(+)-ATPase beta-subunits are somehow different from each other and are named the NK-type and KH-type, respectively. The extracellular domain of the beta-subunit is involved in the folding of the alpha-subunit leading to trypsin-resistant conformations. The sequences from Cys150 to the COOH-terminus of the Na+/K(+)-ATPase beta-subunit and from Ile89 to the COOH-terminus of the H+/K(+)-ATPase beta-subunit are necessary to form trypsin-resistant conformations of the NK- and HK-type, respectively. The first disulfide loop of the extracellular domain of the beta-subunits is critical in the expression of functional Na+/K(+)-ATPase. PMID- 9217966 TI - The GS-X pump in plant, yeast, and animal cells: structure, function, and gene expression. AB - This review addresses the recent molecular identification of several members of the glutathione S-conjugate (GS-X) pump family, a new class of ATP-binding cassette (ABC) transporters responsible for the elimination and/or sequestration of pharmacologically and agronomically important compounds in mammalian, yeast and plant cells. The molecular structure and function of GS-X pumps encoded by MRP, cMOAT, YCF1, and AtMRP genes, have been conserved throughout molecular evolution. The physiologic function of GS-X pumps is closely related with cellular detoxification, oxidative stress, inflammation, and cancer drug resistance. Coordinated expression of GS-X pump genes, e.g., MRP1 and YCF1, and gamma-glutamylcystaine synthetase, a rate-limiting enzyme of cellular glutathione (GSH) biosynthesis, has been frequently observed. PMID- 9217967 TI - Characterisation of the role of calcium in AlF4-induced long-term potentiation in area CA1 of rat hippocampus. AB - We investigated calcium influx in the long lasting potentiation induced in area CA1 of rat hippocampus by brief bath application of the G-protein activator A1F4 (NaF/AlCl3). Brief (10 min) bath application of A1F4 in standard saline (with 2 mM Ca2+) consistently induced a long lasting potentiation which was not observed if A1F4 was bath-applied in nominally calcium free saline. Increasing the potential calcium influx, either by raising extracellular calcium concentration to 3.5 mM or by addition of the voltage operated calcium channel (VOCC) agonist BayK8644, failed to increase the number of slices exhibiting potentiation or the mean level of potentiation. Bath application of AlF4 in the presence of the VOCC antagonist failed to block the potentiation and A1F4- readily induced a long lasting potentiation under voltage clamp conditions, strongly suggesting that the calcium influx required for A1F4-induced potentiation is not through NMDA receptors or VOCC channels. It is suggested that the calcium required may be provided by an ongoing recharging and emptying of IP3 sensitive intracellular Ca2+ stores. PMID- 9217968 TI - Generation of H2O2 by human neutrophils and changes of cytosolic Ca2+ and pH of rat thymocytes in response to galactoside-binding proteins (lectins or immunoglobulins). AB - In contrast to plant agglutinins, biological activities of animal/human lectins are not well defined yet. Testing a panel of seven mammalian carbohydrate-binding proteins we have found that the dimeric lectin from chicken liver (CL-16) was a stimulator of H2O2 release from human neutrophils as well as effector for induction of cytosolic Ca2+ and pH increase in rat thymocytes. Activity of this lectin was comparable to potent galactoside-specific plant lectins such as Viscum album L. agglutinin. The activities of the tested plant lectins depended significantly on their nominal carbohydrate specificity as well as on the source. The results indicate that endogenous lectins may be involved in the regulation of neutrophil and lymphocyte functions by elicitation of selective biosignaling reactions. PMID- 9217969 TI - Vitamin E decreases the order of the phospholipid model membranes in the gel phase: an FTIR study. AB - The effect of alpha-tocopherol on the frequency of the CH2 stretching bands of infrared spectra of dipalmitoylphosphotidylcholine multibilayers has been investigated, both in H2O and 2H2O buffer, to determine the reason for the discrepancy in the literature between the results of different spectroscopic techniques relating to the effect of alpha-tocopherol on membrane order in the gel phase. In contrast to previous FTIR studies, the present FTIR results indicate that alpha T increases the frequencies of the CH2 stretching bands in the gel phase, which implies an increase in the number of gauche conformers (increase in disordering), which is in agreement with other ESR and NMR spectroscopic studies. PMID- 9217970 TI - Neuropathogenesis of chimeric simian/human immunodeficiency virus infection in pig-tailed and rhesus macaques. AB - We recently reported that a chimeric simian/human immunodeficiency virus (SHIVKU 1) developed in our laboratory caused progressive depletion of CD4+ T lymphocytes and AIDS within 6 months of inoculation into pig-tailed macaques (M. nemestrina). None of the pig-tailed macaques showed productive SHIV infection in the central nervous system (CNS). In this report, we show that by further passage of the pathogenic virus in rhesus macaques [M. mulatta], we have derived a new strain of SHIV (SHIVKU-2) that has caused AIDS and productive CNS infection in 3 of 5 rhesus macaques infected with the virus. Productive replication of SHIV in the CNS was clearly shown by high infectivity titers and p27 protein levels in brain homogenates, and in 2 of the 3 rhesus macaques this was associated with disseminated, nodular, demyelinating lesions, including focal multinucleated giant cell reaction, largely confined to the white matter. These findings were reminiscent of HIV-1 associated neurological disease, and our immunohistochemical and in situ hybridization data indicated that the neuropathological lesions were associated with the presence of SHIV-specific viral antigens and nucleic acid respectively. However, the concomitant reactivation of opportunistic infections in these macaques suggested that such pathogens may have influenced the replication of SHIV in the CNS, or modified the neuropathological sequelae of SHIV infection in the rhesus species, but not in pig-tailed macaques. Our findings in the two species of macaques highlight the complexities of lentiviral neuropathogenesis, the precise mechanisms of which are still elusive. PMID- 9217972 TI - Association of EGFR gene amplification and CDKN2 (p16/MTS1) gene deletion in glioblastoma multiforme. AB - Glioblastoma multiforme (GBM) can be divided into genetic subsets: approximately one-third of GBM, primarily in older adults, have EGFR amplification; another one third, primarily in younger adults, have TP53 mutation. The majority of GBM also have homozygous deletions of the CDKN2 (p16/MTS1) gene, resulting in cell cycle deregulation and elevated proliferation indices. We evaluated the relationship between CDKN2 deletions and the GBM subsets as defined by EGFR amplification or TP53 mutation in 70 GBM. Twenty-eight cases (40%) had EGFR amplification, 21 (30%) had TP53 mutation, and 21 (30%) had neither change. CDKN2 deletions were present in 36 (51%) GBM. Of the 28 GBM with EGFR amplification, 20 (71%) had CDKN2 deletion (p = 0.0078). The remaining 16 cases with CDKN2 loss were divided between GBM with TP53 mutations (6 cases) and GBM with neither EGFR amplification nor TP53 mutation (10 cases). Thus, CDKN2 deletions occur twice as commonly in GBM with EGFR amplification (71%) than in GBM with TP53 mutation (29%). CDKN2 deletions occurred in GBM from patients somewhat older than those patients with GBM lacking CDKN2 deletion (mean age 53 vs. 48 years). Specifically among GBM with EGFR amplification, those with CDKN2 deletions also occurred in patients slightly older than those few GBM without CDKN2 deletions (mean age 55 vs. 51 years). The presence of CDKN2 deletions in most GBM with EGFR amplification and in generally older patients may provide one explanation for the potentially more aggressive nature of such tumors. PMID- 9217971 TI - Fas ligand expression in glioblastoma cell lines and primary astrocytic brain tumors. AB - Fas/APO-1 (CD95) is a cell surface receptor that mediates apoptosis when it reacts with Fas ligand (FasL) or Fas antibody. We previously reported that Fas expression is predominantly induced in perinecrotic glioma cells, suggesting that Fas induction is associated with apoptosis and necrosis formation, a histological hallmark of glioblastomas. In this study, we assessed the expression of FasL in 10 glioblastoma cell lines and in 14 astrocytic brain tumors (three low-grade astrocytomas and 11 glioblastomas). Reverse transcriptase (RT)-PCR revealed that all glioblastoma cell lines and primary astrocytic brain tumors express FasL. Immunohistochemically, FasL was predominantly expressed on the plasma membrane of glioma cells. These results suggest that FasL expression is common in human astrocytic brain tumors and may cause apoptosis of glioma cells if Fas expression is induced. PMID- 9217973 TI - Inherited neurodegenerative disorders caused by CAG/polyglutamine tract expansions: symposium introduction. PMID- 9217974 TI - Clinical aspects of CAG repeat diseases. AB - Seven neurodegenerative disorders are known to be caused by unstable expansions of the trinucleotide CAG within human genes, and more will be discovered in the coming years. These disorders share some clinical similarities, as well as some differences, which are summarized here. These diseases have unusual clinical genetic properties related to the dynamic nature of CAG repeat expansions, including instability of the repeat expansion in meiosis, particularly male meiosis; a strong correlation between onset age and size of the repeat expansion; anticipation (earlier disease onset in succeeding generations); new mutations arising from unstable, mutable alleles with a high-normal CAG repeat number; and reduced penetrance for alleles in the low-affected range. Much more remains to be learned about the molecular biology and clinical pathophysiology of this new class of genetic diseases. PMID- 9217975 TI - The neuropathology of CAG repeat diseases: review and update of genetic and molecular features. AB - Classification of inherited neurodegenerative diseases is increasingly based on their genetic features, which supplement, clarify, and sometimes replace the older clinical and pathologic schemata. This change has been particularly rapid and impressive for the CAG repeat disorders. In Huntington's disease, X-linked spinobulbar muscular atrophy, dentatorubropallidoluysian atrophy, and a series of autosomal dominant cerebellar atrophies, genetic advances have resolved many nosologic issues, and opened new avenues for exploration of pathogenesis. In this review, we summarize classic and current concepts in neuropathology of these CAG repeat diseases. PMID- 9217976 TI - The CAG/polyglutamine tract diseases: gene products and molecular pathogenesis. AB - In the past few years, a new type of genetic mutation, expansion of trinucleotide repeats, has been shown to cause neurologic disease. This new class of mutations was first identified in 1991 as the underlying genetic defect in spinal and bulbar muscular atrophy and the fragile X syndrome, and in recent years, trinucleotide repeat expansions have been found to be the causative mechanism in 10 other neurologic diseases. These mutations are produced by heritable unstable DNA and are termed "dynamic mutations" because of changes in the number of repeat units inherited from generation to generation. In the normal population, these repeat units, although polymorphic, are stably inherited. To date four types of trinucleotide repeat expansions have been identified: (1) long cytosine-guanine guanine (CGG) repeats in the two fragile X syndromes (FRAXA and FRAXE), (2) long cytosine-thymine-guanine (CTG) repeat expansions in myotonic dystrophy, (3) long guanine-adenine-adenine repeat expansions in Friedreich's ataxia and (4) short cytosine-adenine-guanine repeat expansions (CAG) which are implicated in eight neurodegenerative disorders and are the focus of this review. Diseases that are caused by trinucleotide repeat expansions exhibit a phenomenon called anticipation that can not be explained by conventional Mendelian genetics. Anticipation is defined as increase in the severity of disease with an earlier age of onset of symptoms in successive generations. Anticipation is often influenced by the sex of the transmitting parent, and for most CAG repeat disorders, the disease is more severe when paternally transmitted. The severity and the age of onset of the disease have been correlated with the size of the repeats on mutant alleles, with the age of onset being inversely correlated with the size of the expansion. In all eight disorders caused by CAG repeat expansion, the repeat is located within the coding region of the gene involved and in all cases it is translated into a stretch of polyglutamines in the respective proteins. All the proteins are unrelated outside of the polyglutamine stretch and most are novel with exception of the androgen receptor and the voltage gated alpha 1A calcium channel, which are mutated in spinal and bulbar muscular atrophy and spinocerebellar ataxia type 6. It is intriguing that the proteins are ubiquitously expressed in both peripheral and nervous tissue but in each disorder only a select population of nerve cells are targeted for degeneration as a consequence of the expanded CAG repeat. Current thinking among scientists working on the molecular mechanisms of neurodegeneration in these diseases is that the presence of an expanded polyglutamine confers a gain of function onto the involved protein. To understand the mechanisms underlying the pathogenesis of these diseases, investigators have turned to generating transgenic mice which recapitulate some of the features of the human disease and hence are excellent model systems to study the progression of the disease in vivo. PMID- 9217977 TI - Trinucleotide repeat instability: genetic features and molecular mechanisms. AB - Trinucleotide repeat expansions are an important cause of inherited neurodegenerative disease. The expanded repeats are unstable, changing in size when transmitted from parents to offspring (intergenerational instability, "meiotic instability") and often showing size variation within the tissues of an affected individual (somatic mosaicism, "mitotic instability"). Repeat instability is a clinically important phenomenon, as increasing repeat lengths correlate with an earlier age of onset and a more severe disease phenotype. The tendency of expanded trinucleotide repeats to increase in length during their transmission from parent to offspring in these diseases provides a molecular explanation for anticipation (increasing disease severity in successive affected generations). In this review, I explore the genetic and molecular basis of trinucleotide repeat instability. Studies of patients and families with trinucleotide repeat disorders have revealed a number of factors that determine the rate and magnitude of trinucleotide repeat change. Analysis of trinucleotide repeat instability in bacteria, yeast, and mice has yielded additional insights. Despite these advances, the pathways and mechanisms underlying trinucleotide repeat instability in humans remain largely unknown. There are many reasons to suspect that this uniquely human phenomenon will significantly impact upon our understanding of development, differentiation and neurobiology. PMID- 9217978 TI - Mouse models of human CAG repeat disorders. AB - Expansions of CAG trinucleotide repeats encoding glutamine have been found to be the causative mutations of seven human neurodegenerative diseases. Similarities in the clinical, genetic, and molecular features of these disorders suggest they share a common mechanism of pathogenesis. Recent progress in the generation and characterization of transgenic mice expressing the genes containing expanded repeats associated with spinal and bulbar muscular atrophy (SBMA), spinocerebellar ataxia type 1 (SCA1), Machado-Joseph disease (MJD/SCA3), and Huntington's disease (HD) is beginning to provide insight into the underlying mechanisms of these neurodegenerative disorders. PMID- 9217979 TI - Toward understanding the molecular pathology of Huntington's disease. AB - Huntington's Disease (HD) is caused by expansion of a CAG trinucleotide beyond 35 repeats within the coding region of a novel gene. Recently, new insights into the relationship between CAG expansion in the HD gene and pathological mechanisms have emerged. Survival analysis of a large cohort of affected and at-risk individuals with CAG sizes between 39 and 50 repeats have yielded probability curves of developing HD symptoms and dying of HD by a certain age. Animals transgenic for the first exon of huntingtin with large CAG repeats lengths have been reported to have a complex neurological phenotype that bears interesting similarities and differences to HD. The repertoire of huntingtin-interacting proteins continues to expand with the identification of HIP1, a protein whose yeast homologues have known functions in regulating events associated with the cytoskeleton. The ability of huntingtin to interact with two of its four known protein partners appears to be influenced by CAG length. Caspase 3 (apopain), a key cysteine protease known to play a seminal role in neural apoptosis, has also been demonstrated to specifically cleave huntingtin in a CAG length-dependent manner. Many of these features are combined in a model suggesting mechanisms by which the pathogenesis of HD may be initiated. The development of appropriate in vitro and animal models for HD will allow the validity of these models to be tested. PMID- 9217980 TI - Huntington's disease and dentatorubral-pallidoluysian atrophy: proteins, pathogenesis and pathology. AB - Each of the glutamine repeat neurodegenerative diseases has a particular pattern of pathology largely restricted to the CNS. However, there is considerable overlap among the regions affected, suggesting that the diseases share pathogenic mechanisms, presumably involving the glutamine repeats. We focus on Huntington's disease (HD) and Dentatorubral-pallidoluysian atrophy (DRPLA) as models for this family of diseases, since they have striking similarities and also notable differences in their clinical features and pathology. We review the pattern of pathology in adult and juvenile onset cases. Despite selective pathology, the disease genes and their protein products (huntingtin and atrophin-1) are widely expressed. This presents a central problem for all the glutamine repeat diseases how do widely expressed gene products give rise to restricted pathology? The pathogenic effects are believed to occur via a "gain of function" mechanism at the protein level. Mechanisms of cell death may include excitotoxicity, metabolic toxicity, apoptosis, and free radical stress. Emerging data indicate that huntingtin and atrophin-1 may have distinct protein interactions. The specific interaction partners may help explain the selective pathology of these diseases. PMID- 9217981 TI - January 1997--7 year old girl with seizures. AB - A seven year old girl presented with six month history of seizures. An MRI scan showed a cortical lesion in the left temporal lobe which was resected. Neuropathologic examination demonstrated meningioangiomatosis, an unusual hamartomatous condition sometimes associated with neurofibromatosis 2. Approximately 50 cases of meningioangiomatosis have been reported in the literature. These are reviewed and compared to the current case. PMID- 9217982 TI - February 1997--9 year old girl with hydrocephalus. AB - A nine year old girl presented with nausea vomiting and hydrocephalus. Imaging studies demonstrated a large right thalamic mass with areas of cystic change or necrosis. Sections of resected material showed perivascular pseudorosettes, necrosis, endothelial proliferation, numerous mitoses, densely cellular areas and numerous foci with a clear-cell morphology. This case is used to illustrate the features of anaplastic ependymomas and to discuss the relationship of the histologic features to prognosis. PMID- 9217983 TI - March 1997--4 year old girl with ring chromosome 22 and brain tumor. AB - A four year old Caucasian girl with a constitutional ring chromosome 22 abnormality and developmental delay presented with increasing ataxia and a six week history of non-specific symptoms. Imaging studies demonstrated a large third ventricular tumor with apparent involvement of the septum. Microscopic and immunohistochemical studies demonstrated an atypical teratoid/rhabdoid tumor. This tumor is compared and contrasted to peripheral malignant rhabdoid tumors and central primitive neuroectodermal tumors. The role of a putative tumor suppressor gene on the long arm of chromosome 22 in the pathogenesis of these tumors is also discussed. PMID- 9217984 TI - Nonlinear dynamics of epileptic seizures on basis of intracranial EEG recordings. AB - PURPOSE: An understanding of the principles governing the behavior of complex neuronal networks, in particular their capability of generating epileptic seizures implies the characterization of the conditions under which a transition from the interictal to the ictal state takes place. Signal analysis methods derived from the theory of nonlinear dynamics provide new tools to characterize the behavior of such networks, and are particularly relevant for the analysis of epileptiform activity. METHODS: We calculated the correlation dimension, tested for irreversibility, and made recurrence plots of EEG signals recorded intracranially both during interictal and ictal states in temporal lobe epilepsy patients who were surgical candidates. RESULTS: Epileptic seizure activity often, but not always, emerges as a low-dimensional oscillation. In general, the seizure behaves as a nonstationary phenomenon during which both phases of low and high complexity may occur. Nevertheless a low dimension may be found mainly in the zone of ictal onset and nearby structures. Both the zone of ictal onset and the pattern of propagation of seizure activity in the brain could be identified using this type of analysis. Furthermore, the results obtained were in close agreement with visual inspection of the EEG records. CONCLUSIONS: Application of these mathematical tools provides novel insights into the spatio-temporal dynamics of "epileptic brain states". In this way it may be of practical use in the localization of an epileptogenic region in the brain, and thus be of assistance in the presurgical evaluation of patients with localization-related epilepsy. PMID- 9217985 TI - Visual evoked potentials relating to imagery: words for concrete objects versus absolute concepts. AB - We recorded visual evoked potentials (VEPs) elicited with high or low imaginable Chinese characters (HIC or LIC), representing concrete objects or absolute concepts, respectively. A closed circle (CC) acts as control stimulus. These were displayed (at 1.6 degrees visual angle) for 35 ms on a TV monitor. Twenty-one channel VEPs (band-pas filter: 0.05-60 Hz), using balanced non-cephalic electrodes, were recorded from -100 to 924 ms for 11 right-handed male volunteers. The VEPs were analyzed by multivariate analysis of variance (MANOVA) and comparison of topographies at four remarkable peaks (P110, N160, P230 and N320). MANOVA showed significant differences (p < 0.001) for both conditions of channel and stimuli (HIC, LIC or CC). P100 for the CC-VEPs, N160 for the HIC- and LIC-VEPs, P230 for the CC-VEPs, and N320 for the HIC-VEPs were remarkable in the posterior scalp regions. Topographies at P100 and N160 showed no difference between the HIC- and LIC-stimuli. However, those at N320 showed difference between the HIC- and LIC-stimuli over the occipital and posterior temporal areas. Those results suggest that the responses at P100 and N160 might segregate Chinese characters from non Chinese characters. N320 suggested certain processes in imagery on recognizing Chinese characters over the occipital and posterior temporal areas. PMID- 9217986 TI - P300 topography of amplitude/latency correlations. AB - The correlational association from 19 electrode sites between peak amplitude and latency for the P3(00) event-related brain potential (ERP) for n = 80 homogeneous subjects was assessed using a simple auditory discrimination task. The correlation strength varied systematically across scalp topography in different ways for the various ERP components. For the target stimuli, P3 amplitude and latency were negatively correlated and most tightly coupled over the frontal central and right medial/lateral recording sites. In contrast, the N1 produced negative correlations that were strongest over the left and right central/lateral locations; P2 demonstrated a positive correlation that was strongest frontally and centrally; N2 demonstrated a positive correlations that was strongest over the central and parietal sites. ERPs from the standard stimuli produced generally similar patterns for the P3 and P2 components, with only weak or no reliable effects observed for the N1 and N2 potentials. Taken together, the findings suggest that analysis of amplitude/latency correlational relationships can provide information about ERP component generation. Theoretical implications are discussed. PMID- 9217987 TI - Age-related changes in cognitive ERPs of attenuation. AB - This investigation explored developmental changes in passive and effortful components of ERPs associated with a visual attention task in children, adolescents, and adults. The task was a 'go-go' version of a continuous performance task, coupled with a passive attending phase in which the subjects merely watched the stimuli of the task. The three age groups featured a constellation of ERP components that shared the same general morphological appearance and distribution, but differences were seen with respect to latencies and amplitudes. Consistent with other studies, there was an inverse relationship with respect to age and peak latencies of the major passive and effortful components. With respect to peak amplitudes, however, the most impressive changes with age were observed in the passive processing components. For example, the P150 and P250 components presented greater amplitudes in children, whereas the N200 component presented its greatest amplitude in adults. While passive in the sense that their appearances were independent of the 'decision-making' process, these components were found to be upwardly adjustable by effort. The late positive component was found to be a combination of a passive P350 and an effortful P450. The P350 component was judged to be largely passive in character as it was well developed in subjects of all age groups when passively attending to the visual stimuli. There was no marked amplitude difference between the child and adult P450 components, but the components peaked in amplitude later in the children. Finally, the children's ERPs featured a distinct frontal negativity (FN) that was present in the Passive phase, but greatly enhanced during the Effortful phase. This study, as have many others, showed that there are reliable developmental changes in the components of visual ERPs. Therefore, the characteristics of the various components of cognitive ERPs may be effective markers of neurodevelopmental status, especially of those neuronal systems vital to attentional processing and effort regulation. PMID- 9217988 TI - A robust assessment of the NoGo-anteriorisation of P300 microstates in a cued Continuous Performance Test. AB - The Continuous Performance Test (CPT) is successfully applied in clinical routine to evaluate attentional performance. The aim of the present study was to investigate the features of the ERPs related to the conditions of a cued CPT, in particular the Go- and the NoGo-condition demanding either the execution or the inhibition of a prepared motor response. For that purpose, 21-channel-ERPs of ten healthy subjects elicited by the Go, NoGo, primer and distractor cues were analyzed with reference-independent methods. The P300 microstates were identified by means of a data-driven segmentation of the ERPs based on the individual peaks of the Global Field Power (GFP). The topographical assessment of the P300 fields yielded an extraordinarily robust result consisting of a more anterior location of the positive centroid in the NoGo compared to the Go condition in every single subject. In conclusion, this result is an impressive validation of the applied reference-independent spatial analysis which reveals the rapid changes of the brain electrical field configurations related to the execution/inhibition paradigm within the cued CPT. Because of the stability of the NoGo anteriorisation we propose to use this parameter as a topographical standard index, analogous to the amplitude effect between oddball targets and nontargets which defines the classical P300. PMID- 9217989 TI - Computerized EEG topography of normal preadolescent twins--correlating similarity of background activity with genetic relatedness. AB - The influence of genetic relatedness on the similarity degree of topographical EEG parameters was studied in a sample of 26 sets of monozygotic (MZ) and 46 sets of dizygotic (DZ) twins. All 144 subjects were healthy, primary school children, aged 7-15 years, 69 boys and 75 girls. Correlation coefficients were calculated for 50 quantitative EEG parameters of paired values obtained at each of 16 active electrode sites, in four groups of paired tracings: 1. MZ twins, 2. DZ twins, 3. The autocorrelated (A) group formed by correlating the spectral parameters from the same subjects in two different analyzed sequences, 4. The random (R) control group of 1200 unrelated pairs formed from DZ twin pairs. Sets of MZ twins and A group showed the highest degrees of similarity of spectral parameters over all brain areas except for significant differences only for some background features over posterior regions. In contrast, highly significant differences in topographic parameters were evident in comparison of MZ sets with DZ sets, particularly when MZ sets were compared with DZ subsets of opposite sex. Both number and degree of significant differences increased progressively in comparisons with groups 3 vs 2, 1 vs 4, and 3 vs 4. The data gave strong evidence for a complex polygenic determination of normal human EEG topography. PMID- 9217991 TI - The generation and subtraction of sensory expectations within cerebellum-like structures. AB - The generation of expectations about sensory input and the subtraction of such expectations from actual input appear to be important features of sensory processing. This paper describes the generation of sensory expectations within cerebellum-like structures of four distinct groups of fishes: Mormyridae; Rajidae; Scorpaenidae; and Apteronotidae. These structures consist of a sheet like array of principal cells. Apical dendrites of the principal cells extend out into a molecular layer where they are contacted by parallel fibers. The basilar regions of the arrays receive primary afferent input from octavolateral endorgans, i.e., electroreceptors, mechanical lateral line neuromasts, or eighth nerve endorgans. The parallel fibers in the molecular layer convey various types of information, including corollary discharge signals associated with motor commands, sensory information from other modalities such as proprioception, and descending input from higher stages of the sensory modality that is processed by the structure. Associations between the signals conveyed by the parallel fibers and particular patterns of sensory input to the basal layers lead to the generation of a negative image of expected sensory input within the principal cell array. Addition of this negative image to actual sensory input results in the subtraction of expected from actual input, allowing the unexpected or novel input to stand out more clearly. Intracellular recording indicates that the negative image is probably generated by means of anti-Hebbian synaptic plasticity at the parallel fiber to principal cell synapse. The results are remarkably similar in the different fishes and may generalize to cerebellum-like structures in other sensory systems and taxa. PMID- 9217992 TI - A quantitative analysis of passive electrolocation behavior in electric fish. AB - Weakly electric fish of the families Gymnotidae and Hypopomidae (Gymnotiformes) are able to locate the electric discharges from conspecifics or from dipole electrodes, and they demonstrate this by making rapid, well-directed approaches toward these electrical sources. A video tracking system was used to follow the movements of electric fish in a large tank and an analytic method was used for computing the direction and magnitude of the electric field anywhere within the cylindrical test tank. Using a static analysis method, we describe the posture of test fish relative to the electric fields during their approaches to stationary or moving electrical stimuli. Using a dynamic analysis, we examine the movements of the fish including the sign and magnitude of velocity and bending in response to electric fields. Electric fish seek to maintain a zero error angle between their body orientation and the local electric field. They do so by bending their body in the direction of the local electric field. The response has a delay of approximately 0.5 s. Swimming in reverse inverts the direction of the bend. These fish also use 'V-turns' to redirect their swim directions when encountering rapidly-changing electric fields. PMID- 9217990 TI - Phenotypic specification of hindbrain rhombomeres and the origins of rhythmic circuits in vertebrates. AB - This essay considers the ontogeny and phylogeny of the cranial neural circuitry producing rhythmic behaviors in vertebrates. These behaviors are characterized by predictable temporal patterns established by a neuronal network variously referred to as either a pacemaker, neural oscillator or central pattern generator. Comparative vertebrate studies have demonstrated that the embryonic hindbrain is divided into segmented compartments called rhombomeres, each of which gives rise to a distinct complement of cranial motoneurons and, as yet, unidentified populations of interneurons. We now propose that novel rhythmic circuits were innovations associated with the adoption of cardiac and respiratory pumps during the protochordate-vertebrate transition. We further suggest that the pattern-generating circuits of more recent innovations, such as the vocal, electromotor and extraocular systems, have originated from the same Hox gene specified compartments of the embryonic hindbrain (rhombomeres 7-8) that gave rise to rhythmically active cardiac and respiratory circuits. Lastly, we propose that the capability for pattern generation by neurons originating from rhombomeres 7 and 8 is due to their electroresponsive properties producing pacemaker oscillations, as best typified by the inferior olive which also has origins from these same hindbrain compartments and has been suggested to establish rhythmic oscillations coupled to sensorimotor function throughout the neuraxis of vertebrates. PMID- 9217993 TI - Biogenic amines and aggression: experimental approaches in crustaceans. AB - This review summarizes our experimental approaches attempting to link amines and their metabolites to aggression in crustaceans. The results demonstrate (i) that agonistic behavior in crustaceans can be quantified, (ii) that the amines themselves have telling and subtle effects on the fighting behavior of animals, (iii) that pharmacological interventions are possible that might allow a biochemical dissection of the underlying mechanisms involved in processes like decision making in these animals, and (iv) that selective metabolites of amines are excreted in the urine of lobsters where they may serve behavioral roles. Many of the studies presented here are preliminary. Nonetheless, we believe the results are provocative and nicely complement previous detailed physiological, morphological and biochemical studies exploring the roles of amines in aggression in crustaceans. We expect that the continued use of this invertebrate model system will allow us to gain considerable insight into, and understanding of, the role served by biogenic amines in a complex behavioral process like aggression. PMID- 9217994 TI - Neural mechanisms of behavioral plasticity: metamorphosis and learning in Manduca sexta. AB - This review summarizes our current understanding of the neural circuit underlying the larval proleg withdrawal reflex (PWR) of Manduca sexta and describes how PWR function changes in two contexts: metamorphosis and learning. The first form of PWR plasticity occurs during the larval-pupal transformation, when the reflex is lost. One mechanism that contributes to this loss is the weakening of monosynaptic excitatory connection from proleg sensory neurons to proleg retractor motor neurons. This change is associated with the hormonally-mediated regression of proleg motor neuron dendrites, which may break synaptic contacts between the sensory and motor neurons. After pupation, some of the proleg motor neurons die in a segment-specific pattern that persists even after individual motor neurons are isolated from the nervous system and exposed to hormones in vitro. The second form of PWR plasticity involves short-term, activity-dependent changes in neural function during the larval stage. The nicotinic cholinergic connections from proleg sensory neurons to motor neurons exhibit several forms of plasticity including facilitation, depression, post-tetanic potentiation and two types of muscarinic modulation. Larval PWR behavior exhibits two simple forms of learning-habituation and dishabituation-which involve alterations in the central PWR circuit. These studies of a simple circuit illustrate neural mechanisms by which behaviors undergo both short- and long-term modifications. PMID- 9217995 TI - In celebration of the life of Walter F. Heiligenberg. PMID- 9217997 TI - Expression of human O6-methyl guanine methyl transferase (MGMT) in post replication repair (PRR) deficient CHO-UV-1 cells: compensation for hypersensitivity to methylating and ethylating agents but not to mitomycin C. AB - The cDNA for human MGMT was transfected into and expressed in CHO cells and the post-replication repair deficient mutant CHO-UV-1 cell, both of which are devoid of endogenous MGMT activity. Expression of MGMT activity was demonstrated by measurement of activity and by immunoblot analysis. The mutant phenotype of UV-1 is characterized by extreme hypersensitivity to killing by methylating and ethylating agents as well as the antitumor antibiotic mitomycin C (MMC). MGMT expression conferred equivalent, supra-normal levels of resistance to killing by MNNG (N-methyl-N'-nitro-nitrosoguanidine) or EMS (ethyl methanesulfonate) on CHO and UV-1, but had no effect on the lethality of MMC. So, even though a mutated gene other than MGMT is known to underlie the pleiotropic phenotype of UV-1, expression of MGMT compensates for part of this phenotype. This result indicates that attempts to concordance map and clone the gene(s) responsible for the UV-1 phenotype can be complicated when using MNNG selection due to compensation by the MGMT gene. These results also indicate that the post-replication repair deficient phenotype characterized in CHO-UV-1 cells, will be masked in cells normally expressing MGMT due to MGMT-mediated resistance to methylating and ethylating agents. PMID- 9217998 TI - Evidence that resistance to osmotic stress is mediated by gene amplification. AB - The mechanism for resistance to osmotic stress in V79 Chinese hamster cells is unknown. Previous experiments have provided little or no support for mechanisms based on gene mutation or stable changes in gene expression. An alternative explanation, based on gene amplification is offered in the present study. The evidence comes from experiments with PALA, which is N(Phosphonoacetyl)L aspartate. Since no other mechanism than gene amplification is known for resistance to PALA, PALA resistance can be used as a reporter for gene amplification in other systems if cross-resistance occurs. Clonal lines resistant to hypertonic NaCl or to hypertonic mannitol were cross-resistant in dose response tests with graded PALA. PALA resistant lines were also obtained by direct exposure of sensitive stock V79 cells to appropriate levels of PALA. In subsequent tests these lines were found to be refractory to osmotic stress when exposed to hypertonic NaCl or hypertonic mannitol. These results suggest that PALA and osmotic stress act as inductors as well as selective agents. The induction of gene amplification is not confined to the system under selection, but occurs at other points in the genome. It is this more general induction that explains cross-resistance between two otherwise unrelated characteristics. PMID- 9217996 TI - Complementation group assignments in Fanconi anemia fibroblast cell lines from North America. AB - Fanconi anemia is a rare autosomal recessive disease characterized by developmental defects of the thumb and radius, childhood onset of pancytopenic anemia and increased risk of leukemia. At least five complementation groups (A-E) have been defined but only the FAC gene has been cloned. Cells can be assigned to complementation group C by direct mutation analysis. To facilitate the search for additional FA genes and to measure the frequency of complementation groups, we have established new genetically marked immortalized FA-A and FA-D fibroblast cell lines and show their usefulness as universal fusion donors. These reference FA cell lines facilitated somatic cell fusion analysis and enabled us to assign the complementation group in 16 unrelated FA patients from North America. The majority of patients, belong to FA complementation group A (69%), followed by FA C (18%), FA-D (4%) and FA-B or FA-E (9%). PMID- 9217999 TI - A respiration-deficient Chinese hamster cell line with a defect in mitochondrial protein synthesis: rapid turnover of some mitochondrial transcripts. AB - Translational initiation and elongation in mammalian mitochondria is still poorly understood, and genetic approaches are expected to be helpful in the elucidation of the mechanism. This study describes a further characterization of a Chinese hamster mutant cell line which is severely defective in mitochondrial protein synthesis. Additional proof is provided that the mutation is nuclear. It is shown that there is no dramatic depletion of mitochondrial DNA in these cells, and the mtDNA appears to have neither significant deletions nor rearrangements. Transcription is not affected, and the polycistronic transcripts are processed normally. However, mt mRNAs are differentially quite unstable, and some are at very low steady state levels. Several nuclear transcripts encoding mitochondrial proteins were also investigated and found to be present at normal levels, and in particular, it was demonstrated that complex II (and succinate dehydrogenase activity), encoded entirely by nuclear genes, was only moderately affected by the absence of all the other complexes of the electron transport chain. PMID- 9218000 TI - Site-directed substitution of Ser1406 of hamster CAD with glutamic acid alters allosteric regulation of carbamyl phosphate synthetase II. AB - Ser1406 of the allosteric region of the hamster CAD enzyme, carbamyl phosphate synthetase II (CPSase), is known to be phosphorylated in vitro by cAMP-dependent protein kinase (PKA). Metabolic labeling experiments described here demonstrate that CAD is phosphorylated in somatic cells in culture. Phosphorylation is stimulated by treating cells with 8-bromo-cAMP, a PKA activator. The stimulation is essentially prevented by pretreatment with H-89, a PKA specific inhibitor. Substitution of Ser1406 with alanine results in an enzyme with kinetics and allosteric regulation indistinguishable from unsubstituted CAD. However, substitution to glutamic acid increases CPSase activity by reducing the apparent Km (ATP). The UTP concentration required to give 50% inhibition is increased rendering this altered enzyme significantly less sensitive to feedback inhibition, but allosteric activation by PRPP is unaffected. While these data do not prove that Ser1406 is phosphorylated in vivo, they do indicate that a specific alteration at this residue can affect allosteric regulation. PMID- 9218001 TI - A role for certain mouse Aprt sequences in resistance to toxic adenine analogs. AB - A mouse embryonal carcinoma cell line hemizygous for the adenine phosphoribosyltransferase gene (aprt) was exposed to ultraviolet light (UV) or to the alkylating agent, ethyl methanesulfonate (EMS). Thirty eight cell lines retaining the aprt gene were isolated by selecting for resistance to 2,6 diaminopurine (DAP), an adenine analogue which selects against aprt activity. Of these, six cell lines distinguished by significant levels of aprt enzymatic activity after selection in DAP, were found to carry mutations in the aprt gene affecting the apparent Km of the enzyme for adenine in every cell line, and the apparent Km for phosphoribosylpyrophosphate in two of the six cell lines. The results indicate that the ability of these cells to survive in the presence of toxic adenine analogues while maintaining significant levels of aprt enzyme activity may be due to a reduced affinity for the adenine analogue, DAP. This biochemical analysis along with results obtained from sequencing the aprt gene from 31 DAP resistant cell lines with no detectable aprt activity were used to implicate certain amino acids within aprt in substrate binding. It was also determined that, in contrast to UV, EMS did not appear to exhibit any strand bias in the distribution of mutations. PMID- 9218002 TI - Isolation of DNA markers for the rat Sai 1 gene for suppression of anchorage independence by using representational difference analysis. AB - We have applied the representational difference analysis (RDA) to isolate genetic markers for a deletion on the rat chromosome RNO5q22-33. This deletion occurred in anchorage independent sublines of a normal rat fibroblast x mouse hepatoma cell hybrid (BS181) (Islam 1989). Normal rat tissue DNA provided the "tester" and the BS181 hybrid DNA the "driver" in the RDA hybridization/selection reactions. Out of twelve RDA derived DNA sequences that were analyzed in detail using a rat X mouse cell hybrid panel for chromosome mapping, nine (75%) were found to represent RNO5 deletions, whereas the other three were new RFLPs mapping to other chromosomes. In two cases, the RDA sequences were also analyzed by fluorescence in situ hybridization (FISH) and found to give distinct signals in the RNOq22-33 region. This result emphasizes teh significance of the previous cytogenetic analysis of this hybrid, which indicated the presence of a gene for the suppression of anchorage independence, Sai 1, in this deletion region. The RDA derived sequences isolated by this work will provide a valuable source of new genetic markers for the further detailed analysis of the Sai 1 deletion region. PMID- 9218003 TI - Homologous junctions formed between a vector and human genomic repetitive LINE-1 elements as a result of one-sided invasion. AB - Studies on homologous recombination in mammalian cells between an exogenous DNA molecule containing a double-strand break and a homologous genomic sequence have indicated that there were at least two distinct types of homologous recombination processes, one that involved the formation of two homologous junctions and another that involved the formation of one homologous junction and one illegitimate junction. Both types of events are produced in gene targeting experiments. We have proposed a model to account for the later process called one sided invasion. One-sided invasion has now been reported in numerous species belonging to different phyla and appears to be a universal mechanism. It has also been observed in normal human germ cells. The role of one-sided invasion is still unknown. Using a recombination assay between LINE-1 elements from the human genome and exogenous LINE-1 sequences, we have characterized the process of homologous junction formation in one-sided invasion. We found that at each of the homologous junctions, variable lengths of the vector L1 sequences had been replaced by genomic L1 sequences. We also found a homologous junction that involved three partners, suggesting that the homologous end could be released and become available for a second round of interaction. PMID- 9218004 TI - Serum markers of bone metastases in postmenopausal breast cancer patients treated with formestane. AB - Bone metabolism marker evaluation is expected to play an auxiliary role in the diagnosis and follow-up of bone metastases in patients affected by different types of neoplasms. In this study we have evaluated osteoblastic and osteoclastic markers in 18 patients with bone metastases from breast cancer at diagnosis and for 1 year of follow-up during treatment with the aromatase inhibitor formestane. Osteoblastic markers include the carboxy-terminal propeptide of type I procollagen, the bone-specific alkaline phosphatase and the bone GLA protein. The carboxy-terminal cross-linked telopeptide of type I collagen (ICTP) was evaluated as a marker of osteoclastic activity. The patients were classified into three groups according to clinical response. A good correlation between marker level modifications and clinical evolution of skeletal metastases was observed for all the examined markers. Patients with progressive disease showed increasing levels of all markers, whereas patients in regression showed a reduction compared to the basal levels; patients with stable disease fell in between these two categories. We also found that basal ICTP values have prognostic significance: in the stable and progressive disease group they were higher than in the partial response group. PMID- 9218005 TI - Expression of E-cadherin and alpha- and beta-catenin mRNAs in uterine cervical cancers. AB - To show the mRNA expressions of E-cadherin and alpha- and beta-catenin-which mainly compose the adherens junction-associated with invasion and metastasis of uterine cervical cancers, we studied the expression of E-cadherin and alpha- and beta-catenin mRNAs in cancers in comparison with normal counterparts. The integral expression of E-cadherin and alpha- and beta-catenin mRNAs was suppressed in the metastatic lesions of advanced uterine cervical cancers, while it was not in the primary tumors. Therefore, the suppressed expression of main adhesion molecules in the adherens junction might contribute to adherens junctional dysfunction, which might lead to invasiveness and metastatic potential of advanced uterine cervical cancers as one rate-limiting step. PMID- 9218006 TI - Tumoricidal activity of antiseptics with assessment of cell viability in mice with severe combined immunodeficiency. AB - Previously, we have assessed the efficacy of different cytotoxic agents on the viability of SW620 human colonic carcinoma cells in vitro. In this study, we have investigated the tumoricidal efficacy of some antiseptic agents on SW620 human colonic carcinoma cells which were subsequently inoculated into severe combined immunodeficient (SCID) mice. An inoculum of 5 x 10(6) cells suspended in 200 microliters buffer solution was found to be the minimum number of cells needed to result in tumour growth within 8 weeks after subcutaneous injection into SCID mice. The integrity of the cells was assessed in vitro with the trypan blue exclusion test after 30 min incubation in distilled water (DW), chloramine 0.5% in DW and polyvinyl pyrrolidone iodine (PVP-I) 0.01, 0.05, 0.1 and 5% in DW. DW and PVP-I 0.01% were not tumoricidal, neither in vitro nor in vivo. In contrast, PVP-I 5% and chloramine 0.5% 'killed' all or almost all tumour cells in vitro and prevented their growth in vivo. PVP-I 0.05 and 0.1% were less effective in vitro than 5%, but could prevent in vivo proliferation unless an adjustment of the residual number of viable tumour cells was performed. These data indicate the importance of the amount of the tumour inoculum and hence the need to use a maximally effective 'killing' agent. PMID- 9218007 TI - Regulation of gelatinase production and invasiveness by organ-specific fibroblasts in high- and low-metastatic clones from murine RCT sarcoma. AB - Regulation of gelatinase production, invasiveness and migration activity by organ specific fibroblasts from embryo, subcutaneous and lung tissues of mice were investigated in high-metastatic RCT+ and low-metastatic RCT- clones established from a poorly differentiated murine sarcoma. In the conditioned media of RCT+ cells, mouse skin fibroblasts (MSF) obtained from the tissue of tumor origin (orthotopic) stimulated the production of the 105-kD gelatinase more than C3H/ 10T1/2 clone 8 (C3H/10 T1/2 CL8) or mouse lung fibroblasts (MLF). In the conditioned media of RCT- cells, however, cocultivation with fibroblasts showed only slight stimulatory effects on the production of the 105-kD gelatinase. In the invasion assay, using a reconstituted basement membrane (matrigel), RCT+ cells cocultivated with MSF showed significantly higher invasiveness than those cocultivated with C3H/10T1/2 CL8 or MLF. However, no significant differences were shown in the invasiveness of RCT- cells in cocultivation with three types of fibroblasts and in cultivation without fibroblasts. There was no significant difference in migration activity between RCT+ and RCT- cells cultivated alone. But in the cocultivation of both clones with MSF, the migration activity of RCT+ cells was significantly higher than that of RCT- cells. These findings suggest that MSF might delineate the difference in characteristics related to the metastatic potential of RCT+ and RCT- cells through regulation by organ-specific factors. PMID- 9218008 TI - Effect of paclitaxel (Taxol) on CA 125 expression and release by ovarian cancer cell lines. AB - Two ovarian cancer cell lines, OVC432 and the newly established CVU4I, were used to study the effect of Taxal on cell growth and simultaneous CA 125 antigen expression. Growth of both cell lines was effectively inhibited by drug concentrations of 0.1 microM and higher. Complete inhibition of cell growth may result from high concentrations of Cremophor EL present in the Taxol formulation. Immunohistochemical analysis demonstrated that both cell lines retained the CA 125 expression on the cell surface during exposure to paclitaxel. This was reflected in a constant statistically significant correlation between cell numbers and CA 125 concentrations found in cell lysates. CA 125 levels in the culture medium showed a significant relation to cell numbers and, consequently, to the response of the cell line to the administered anticancer drug. It may be concluded from this study that CA 125 seems to be a reliable tumor marker in monitoring tumor response during paclitaxel treatment. PMID- 9218009 TI - Prognostic significance of tumour markers in endometrial cancer. AB - Serum cancer antigen (CA) 125, CA15.3, CA19.9, carcinoembryonic antigen and tissue polypeptide antigen were analyzed in 100 normal subjects, 47 patients with benign gynaecological diseases and 97 patients with endometrial cancer. The incidence of individual elevated tumour markers (> 2SD) was 21.5-30.9% in cancer patients. Elevations of CA125 and CA15.3 were significantly associated with poor prognostic clinical factors. Univariate anaylses showed that elevated CA125, CA15.3 and CA19.9 were significantly associated with shorter survival. In multivariate analysis, CA15.3 was highly significant and had a larger hazard ratio. In conclusion, CA15.3 is a useful marker for the prognosis of patients with endometrial cancer. PMID- 9218010 TI - Hyperthermia potentiates antitumor effect of thermosensitive-liposome encapsulated melphalan and radiation in murine melanoma. AB - Malignant melanoma are chemoresistent tumors with poor prognosis. The aim of this study was to determine whether multimodality therapy of murine melanoma involving a combination of radiation with thermosensitive-liposome-encapsulated melphalan and local hyperthermia would result in enhancement of therapeutic efficacy for a more effective management of melanoma. Melphalan was entrapped in thermosensitive liposomes prepared from natural lipids: egg phosphatidyl choline, cholesterol and ethanol to show phase transition at 42 +/- 0.5 degrees C and used in combination with localized heating of B16F10 murine melanoma transplanted into the legs of C57B1/6 mice for selective drug targeting at the tumors and/or radiation for treatment of melanoma. Murine melanoma transplanted into C57B1/6 mice were subjected to bimodality treatments involving a combination of radiation, hyperthermia or melphalan. Partial tumor regression was observed in mice receiving a combination of hyperthermia and radiation (median tumor volume 427.3 mm3) or a combination of free melphalan and radiation (512.1 mm3) as compared to untreated controls (630.9 mm3). Each group consisted of 18 animals, and the results are expressed as median tumor volume +/- SD. Animals receiving multimodality therapy comprising irradiation followed by injection of thermosensitive liposomal melphalan and hyperthermic treatment of the tumor bearing leg at 42 +/- 0.5 degrees C for 1 h showed marked tumor regression in comparison with untreated controls or animals treated with a combination of radiation and hyperthermia or radiation and free-drug melphalan. Animals receiving thermoradiochemotherapy also showed prolonged survival; 70% of animals survived for more than 3 months. The study shows greater tumor cell killing, tumor growth delay and prolonged survival produced by a combination of radiation, thermosensitive-liposome-entrapped melphalan and hyperthermia compared with animals receiving single-modality or bimodality treatments. It is concluded that this multimodality approach will be potentially useful for more effective management of melanoma. PMID- 9218011 TI - Citrate metabolism of normal and malignant prostate epithelial cells. PMID- 9218012 TI - Biologic variability of prostate-specific antigen and its usefulness as a marker for prostate cancer: effects of finasteride. The Finasteride PSA Study Group. AB - OBJECTIVES: The effects of finasteride on prostate-specific antigen (PSA) variability and usefulness in prostate cancer detection were examined. METHODS: Percent change and crossover of PSA levels between the low (1.0 to 3.9 ng/mL) and high (4.0 to 10.0 ng/mL) ranges were evaluated in 72 men with benign prostatic hyperplasia (BPH) and 77 men with both BPH and prostate cancer (PCa) treated with finasteride or placebo for 6 months. Patients with PCa were studied as a model for evaluating the effects on PSA levels in patients with BPH and latent PCa. As recommended on the product label, PSA levels for finasteride-treated patients were doubled for interpretation. RESULTS: In patients with BPH, most placebo- and finasteride-treated patients with low PSA levels at baseline had subsequent PSA levels below 4.0 ng/mL throughout the study. Among patients with high baseline PSA levels, only 1 of 17 finasteride-treated patients compared with 8 of 13 placebo-treated patients crossed into the low range. In the BPH/PCa study, most placebo-treated patients maintained PSA levels in the same range (15 of 19 less than 4.0 ng/mL; 14 of 16 greater than 4.0 ng/mL). Almost one third of finasteride treated patients with low PSA levels at baseline crossed into the high range (8 of 22), whereas most patients with high PSA levels at baseline were not masked with treatment, with PSA levels remaining high (12 of 15). CONCLUSIONS: PSA levels cross between the low and high PSA ranges in both finasteride- and placebo treated patients with BPH and those with both BPH and PCa. Doubling the PSA levels in finasteride-treated patients allows appropriate interpretation of PSA values and does not mask the detection of PCa. PMID- 9218013 TI - A posterior lumbar approach for retroperitoneoscopic adrenalectomy: assessment of surgical efficacy. AB - OBJECTIVES: To compare the efficacy of retroperitoneoscopic adrenalectomy by a posterior lumbar approach (RPA) with that obtained by a transperitoneal anterior approach (TAA) or retroperitoneal lateral flank approach (RLA). METHODS: Fifty one patients underwent endoscopic adrenalectomy by three approaches, including laparoscopic adrenalectomy by TAA in 33, retroperitoneoscopic adrenalectomy by RLA in 5, and retroperitoneoscopic adrenalectomy by RPA in 13. RESULTS: The average adrenal tumor size was 27 mm (range 8 to 65). The average number of trocars required for RPA was 3.2 which was significantly less than that for TAA and for RLA (4.2 and 4.1, respectively). The conversion rate to open surgery was 9.1% by TAA, 0% by RLA, and 7.7% by RPA. The average operating time for TAA was 252 minutes, which was significantly shortened to 194 minutes by RLA and 142 minutes by RPA (P < 0.02). The average blood loss was 101 mL for TAA and was negligible by RLA and RPA (22 and 32 mL. respectively). CONCLUSIONS: RPA allowed direct access to the main adrenal vascular supply before the gland was greatly manipulated. Endoscopic adrenalectomy by TAA or even by RLA required extra ports for retraction of liver, spleen, vena cava, or adrenal gland, with higher chance of vein avulsion. RPA was technically feasible and most effective for retroperitoneoscopic adrenalectomy in regard to the simplicity of vascular control. The operating time, perioperative morbidity, and cost were reduced with this approach. PMID- 9218014 TI - Prospective evaluation of fine needle aspiration of small, solid renal masses: accuracy and morbidity. AB - OBJECTIVES: To determine the accuracy and clinical utility of fine needle aspiration (FNA) of small, solid renal masses. METHODS: A total of 25 patients with small (less than 5.0 cm), solid, clinically localized renal masses were prospectively identified and evaluated with computed tomography guided FNA with analysis for presence of malignant cells and determination of nuclear grade. The final pathologic findings were used for comparison in each case. All patients had renal cell carcinoma and were managed with radical or partial nephrectomy; 3 had low-grade lesions (Fuhrman's grade 1/4), 2 had high-grade lesions (Fuhrman's grade 4/4), and all other patients had intermediate-grade lesions (Fuhrman's grade 2/4 or 3/4) on final histopathologic assessment. RESULTS: Overall, 10 aspirations yielded diagnostic malignant cells, and 9 were read as rare as rare atypical cells suspicious for malignancy. The remainder were negative (n = 6). Correlation with final nuclear grade was observed in eight instances and discordance in two instances. Subcapsular hematomas were observed at the time of surgery in 10 patients, but in no instance was the operation adversely affected. CONCLUSIONS: The diagnostic yield of FNA of small, solid renal masses appears to be too low to justify the potential morbidity of the procedure. PMID- 9218015 TI - Cystic pyeloureteritis: review of 34 cases. Radiologic aspects and differential diagnosis. AB - OBJECTIVES: To refine the clinical and radiologic description of an unusual benign disease, cystic pyeloureteritis (CPU), consisting of the appearance of suburothelial cysts that raise the mucosa layer of the urothelium. We also studied its relationship with various types of inflammation, including chronic infection, that may be the stimulus for the appearance of CPU. METHODS: We compiled 34 cases of CPU covering the period 1976 to 1994, analyzing the clinical manifestations, diagnostic procedures, differential diagnosis, and evolution. RESULTS: There are no specific symptoms associated with the presence of cysts. The average age of the patients was 59 years (range 30 to 77). Urinary tract infection was detected in 18 (53%). The pyeloureteritis was unilateral in 27 (79%) and bilateral in 7 (21%) of the patients. The location of the cysts was as follows: 1 pyelic (3%); 6 pyeloureteral (18%); and 27 (79%) ureteral. Resolution of the radiologic alterations depends on the resolution of the associated pathology: infections, lithiasis, and obstruction. CONCLUSIONS: We conclude that CPU is a benign pathology with indolent evolution and variable duration; it is not associated with sequelae. Diagnosis is made on the basis of radiologic findings, mainly intravenous urography; in view of the minor entity of the pathology, biopsy is not advisable if the radiologic findings are conclusive. PMID- 9218016 TI - Efficacy of pentosan polysulfate in the treatment of interstitial cystitis: a meta-analysis. AB - OBJECTIVES: To determine the efficacy of pentosan polysulfate (Elmiron) compared to placebo in the treatment of interstitial cystitis. METHODS: The data sources used were MEDLINE, Excerpta Medica, and International Pharmaceutical Abstracts databases, and the manufacturer. Bibliographies of articles obtained were reviewed. The keywords used were pentosanpolysulfate, pentosanpolysulfate sodium, and pentosan. Inclusion criteria were blinded selection of English language, prospective, randomized, placebo-controlled comparative trials; > or = 8 weeks' duration; > or = 300 mg daily; adult humans with > or = 1 symptoms including pain, urgency, frequency, and nocturia; symptoms for > or = 12 months; normal urinalysis; negative findings for urine culture and cytology. Exclusion criteria were hemorrhagic cystitis; drug-, microbial-, or radiation-induced cystitis; carcinoma in situ; other influencing diseases. The outcome of success was defined as a > or = 50% decrease in pain, urgency, frequency, and nocturia. The number of successes was extracted by blinded investigators, treating withdrawals as failures. The percentage difference in success rates of pentosan polysulfate and placebo, and the number needed to treat (NNT) were determined for each variable; P values and 95% confidence intervals (CIs) were determined for combined data. Homogeneity of effect was determined by calculating Q (chi-squared). Article quality was assessed using the Chalmers scale to determine if quality affected outcome. Effective inter-rater reliability was determined using Rosenthal's method. Significance was set at P < 0.05. RESULTS: Four studies were included. Data were extracted from all four studies for pain (n = 398), three for urgency (n = 306), two for frequency (n = 160), and one study for nocturia (n = 106). The differences (95% confidence limits) were pain: 16.6% (95% CI 8%, 25%), NNT = 7; urgency: 13.0% (1.0%, 25%), NNT = 7.5; frequency: 16.7% (2.3%, 31.1%), NNT = 6; nocturia: -1% (-19.8%, 21.8%). P values from homogeneity tests were not significant. Mean quality scores were 63.8%, 48.1%, 50.4%, and 65.6%, respectively, in the four studies; the effective inter-rater reliability was 0.96. Results did not differ when weighted by quality score. CONCLUSIONS: Pentosan polysulfate is more efficacious than placebo in the treatment of pain, urgency, and frequency associated with interstitial cystitis. Pentosan polysulfate is not significantly different from placebo in treating nocturia associated with interstitial cystitis. PMID- 9218017 TI - Holmium:yttrium-aluminum-garnet laser cystolithotripsy of large bladder calculi. AB - OBJECTIVES: Patients with large bladder calculi (4 cm or larger) have traditionally been managed with open cystolithotomy. Endoscopic management with cystolitholapaxy or electrohydraulic lithotripsy risks complications. In an effort to spare patients the morbidity of open cystolithotomy, the results of holmium:yttrium-aluminum-garnet (YAG) laser cystolithotripsy for bladder calculi 4 cm or larger were reviewed. METHODS: Consecutive patients with bladder calculi of 4 cm or larger were managed with holmium:YAG laser cystolithotripsy. Laser energy was delivered using either the 365-micron end-firing fiber or the 550 micron side-firing fiber. RESULTS: Fourteen consecutive patients were managed with holmium:YAG cystolithotripsy. All patients were rendered stone free, regardless of stone composition or size. Median anesthesia time was 57 minutes. Twelve of 14 patients were discharged by the first postoperative day. The procedure times normalized for stone size (mean +/- standard deviation) for the end-firing versus the side-firing fibers were 13 +/- 6 min/cm versus 6 +/- 1 min/cm, respectively; P = 0.04. CONCLUSIONS: Holmium:YAG laser cystolithotripsy of large bladder calculi is effective, technically facile, and safe. The 550 micron side-firing fiber may be better suited for large bladder calculi compared with the 365-micron end-firing fiber. Holmium:YAG cystolithotripsy may obviate open cystolithotomy in selected patients. PMID- 9218018 TI - Comparison of the Bard BTA test with voided urine and bladder wash cytology in the diagnosis and management of cancer of the bladder. AB - OBJECTIVES: To compare the Bard BTA test, a simple latex-agglutination test for cancer of the bladder (BC) that can be performed in less than 3 minutes in the urologist's office, with voided urine or bladder wash cytology in the diagnosis of subjects suspected of having BC on the basis of symptoms or recent abnormal cystoscopy or intravenous urography. METHODS: The study was performed at three medical centers in 414 subjects (147 female and 267 male; mean age 60 years), 345 of whom (83%) had no prior history of BC. The cytologic examinations were performed by pathologists unaware of the results of the BTA test. RESULTS: Cystoscopy or cystoscopy and biopsy revealed BC in 71 subjects (17%). The overall sensitivities of the BTA test and cytology were 70% and 25%, respectively. The specificities of the BTA test and cytology in the 337 subjects without BC were 90% and 100%, respectively. The sensitivities of the BTA test by tumor grade were 17%, 64%, and 92% for grades 1, 2, and 3, respectively; those of cytology were 17%, 14%, and 44%. Regression analysis suggests that tumor grade but no other study variable explains the sensitivity of the BTA test. CONCLUSIONS: The BTA test is considerably more sensitive than cytology in the detection of BC. For urologists who use cytology in the diagnosis and follow-up of patients with BC, the BTA test may replace cytology. PMID- 9218019 TI - Urodynamic and clinical effects of noninvasive and minimally invasive treatments in elderly men with lower urinary tract symptoms stratified according to the grade of obstruction. AB - OBJECTIVES: We investigated the symptomatic and urodynamic effects of several noninvasive and minimally invasive treatment modalities to quantify these effects and to compare subjective and objective results within groups with various degrees of obstruction. METHODS: In a prospective study at one center, 487 patients who completed a full screening program including urodynamic investigation started treatment with watchful waiting, terazosin, transurethral microwave thermotherapy, or laser treatment of the prostate; they were re evaluated symptomatically and urodynamically after 6 months of therapy. The symptomatic and urodynamic results of 87 patients from another center who underwent transurethral resection of the prostate and who had their second urodynamic evaluation 6 months after surgery were also included. RESULTS: In patients without bladder outlet obstruction (BOO), improvement in maximum flow and symptom scores with little change in the degree of obstruction was most apparent, whereas a decrease of detrusor pressure at maximum flow was observed mainly in patients with BOO. The urodynamic effect but not the symptomatic effect of treatments depended on the initial grade of BOO. Urodynamic changes were more marked in the minimally invasive treatment groups compared with the noninvasive treatment groups. CONCLUSIONS: In symptomatic patients with benign prostatic hyperplasia, symptomatic improvement in the short term does not seem to depend on changes in urodynamic parameters. Future well-controlled studies focusing on the durability of symptomatic and urodynamic effects will be needed to illustrate the relative potential of urodynamic and other clinical parameters to predict a favorable response to current and innovative treatments. PMID- 9218020 TI - Prospective multicenter ProLase II clinical trial of neodymium:yttrium-aluminum garnet laser prostatectomy. AB - OBJECTIVES: To assess the clinical efficacy of neodymium:yttrium-aluminum-garnet (YAG) laser coagulation prostatectomy using a broad-angle, divergent-beam, side firing fiber. METHODS: Eighty adult men with voiding symptoms caused by benign prostatic hyperplasia were enrolled in a prospective multicenter study of free beam neodymium:YAG laser prostatectomy performed with the ProLase II side-firing delivery fiber. Voiding outcomes were assessed at 3, 6, and 12 months postoperatively. RESULTS: At 1-year follow-up, peak urinary flow rates were increased by 105%, postvoid residual urine volumes had decreased by 38%, and the AUA symptom index had decreased by 60%. Serious treatment-related complications occurred in 3 of 80 patients (3.8%). The reoperation rate through 1-year follow up was 2.7%. CONCLUSIONS: Neodymium:YAG laser prostatectomy performed with the ProLase II delivery fiber has proven safe and efficacious with durable results through 1 year in the relief of symptomatic bladder outlet obstruction due to benign prostatic hyperplasia. PMID- 9218021 TI - Role of PSA and its indices in determining the need for repeat prostate biopsies. AB - OBJECTIVES: To retrospectively study evaluated prostate-specific antigen (PSA) and four PSA indices as criteria for performing a repeat biopsy when the initial biopsy findings are negative for cancer. METHODS: One, two, or more repeat biopsy sessions were performed on 193, 54, and 14 men, respectively, all of whom had an initially negative biopsy at our institution. We compared the usefulness of PSA, PSA density, age-referenced PSA, volume-referenced PSA, and PSA velocity for predicting the presence of cancer. RESULTS: Overall, 51 men (26%) were found to have cancer on repeat biopsy. Cancer was found in 17% (33 of 193) of the men on the first repeat biopsy and 26% (14 of 54) of the men who had a second repeat biopsy. Of all the indices, PSA and volume-referenced PSA had the highest sensitivity, missing the fewest cancers; however, this was achieved at the expense of saving the least number of biopsies. We evaluated the usefulness of indices among 9 patients who had a normal PSA at the initial biopsy; volume referenced PSA was the earliest predictive index indicating that a repeat biopsy be performed. Analysis of the area under the receiver-operating characteristic curves revealed no significant advantage for any index over PSA alone in determining who should undergo a repeat biopsy. CONCLUSIONS: In determining the need for repeat biopsies, the established PSA threshold value of 4 ng/mL is equivalent or superior to age-referenced PSA, volume-referenced PSA, PSA density, and PSA velocity. However, volume-referenced PSA has the potential to be the earliest index predicting cancer in men with normal PSA level. PMID- 9218023 TI - Approaches to prostate cancer by managed care organizations. AB - OBJECTIVES: Managed care organizations (MCOs) are developing population-based approaches to illnesses with large numbers of patients, wide variations in care and outcomes, and high costs. This is the first survey that evaluates current prostate cancer approaches by MCOs. METHODS: Case studies and a survey of corporate medical directors at large MCOs were conducted. RESULTS: Two approaches, broadly based on disease management strategies for men with prostate cancer, have been implemented in managed care settings on the West Coast. While both have provided comprehensive approaches to the disease, assessment of improvement in outcomes will require longer follow-up. A survey of corporate medical directors of MCOs indicates that population-based disease approaches nationwide for malignant prostate disease lag behind more well-developed efforts for nonmalignant illnesses such as diabetes and asthma. CONCLUSIONS: Prostate cancer may be a feasible area for development and evaluation of population-based approaches. MCOs have the potential to improve clinical care and outcomes for large numbers of men with prostate cancer. While limitations exist related to specific managed care considerations of data needs and lack of medical and surgical consensus on disease management, programs based on shared decision making have the potential to improve patient care and outcomes. PMID- 9218022 TI - Re-examining the role of prostate-specific antigen density in predicting outcome for clinically localized prostate cancer. AB - OBJECTIVES: To evaluate the prognostic significance of prostate-specific antigen density (PSAD) in clinically localized prostate cancer and determine whether this index is independent of or superior to prostate-specific antigen (PSA) in predicting outcome of patients treated with external beam radiotherapy. METHODS: Between January 1989 and December 1993, 175 evaluable patients with clinically localized prostate cancer received definitive radiotherapy using computed tomography (CT)-guided conformal techniques. PSAD was defined as the ratio of the pretreatment serum PSA to the prostate volume measured from CT treatment planning scans by one investigator. All PSA values were determined using the Hybritech assay. Biochemical failure was defined as two consecutive elevations in PSA separated by at least 3 months and a final PSA value greater than 1 ng/mL. RESULTS: Multivariate analysis including PSA and Gleason score revealed both to be statistically significant predictors of biochemical disease-free survival (P = 0.048 and P < 0.001, respectively). PSAD did not achieve significance on regression analysis. A direct multivariate analysis including PSA and PSAD required dichotomization in order to reduce high correlation. This analysis demonstrated a relative risk (RR) for failure of 1.27 (NS) for high PSA versus low PSA compared with a RR of 1.20 (NS) for high PSAD versus low PSAD. A regression model containing all three variables indicated only the Gleason score as significant in predicting biochemical failure. CONCLUSIONS: These data do not suggest that PSAD is either an independent prognostic factor or a stronger discriminant of outcome than PSA in patients with clinically localized prostate cancer treated with definitive external beam radiotherapy. Larger patient numbers with longer follow-up data, use of a clinical end point, or an analysis restricted to the appropriate subgroup may demonstrate the utility of PSAD in the future. PMID- 9218024 TI - Conformal external beam treatment of prostate cancer. AB - OBJECTIVES: This study reports the 5-year outcomes of treatment for patients with prostate cancer treated largely with conformal three-dimensional radiation therapy. METHODS: Results are presented for 456 consecutive patients treated prior to December 31, 1993 whose pretreatment prostate-specific antigen (PSA) levels are known. Biochemical failure was defined as two consecutive rises in the PSA that equals or exceeds 1.5 ng/mL. Kaplan-Meier product limit methods, the log rank test, and Cox regression models were used in evaluating the data. No patient was lost to follow-up. RESULTS: The 5-year biochemically free of failure (bNED) rate for all patients was 61% and 57% at 7 years. In the group with pretreatment PSA less than 10 ng/mL, the 5-year bNED rate for patients with localized disease (T1,2AB disease, Gleason sum of 6 or less) was 85% and for those with locally advanced disease (T2C,3), 70%. In the group with pretreatment PSA of 10 to 19.9 ng/mL, the 5-year bNED rate for patients with localized disease was 66% and for those with locally advanced disease, 44%. In the group with pretreatment PSA of 20 ng/mL or above, the patients with localized or locally advanced disease had 5 year bNED rates of 31% and 21%, respectively. CONCLUSIONS: The results of largely conformal three-dimensional external beam treatment of localized prostate cancer produced 5-year bNED results that are comparable to recent reports of nerve sparing prostatectomy. Preliminary 7-year bNED results in all patients and in patients with localized tumors indicated a modest decrease in the cancer-free rate from that observed at 5 years, suggesting the results are durable. PMID- 9218025 TI - Hazard rates for progression after radical prostatectomy for clinically localized prostate cancer. AB - OBJECTIVES: We calculated the annual hazard rate (HR) for prostate cancer recurrence after radical prostatectomy (RP) to elucidate the pattern of treatment failure over time and to assess the efficacy of definitive therapy. METHODS: We calculated the progression-free probabilities (PFP) and HRs after RP for a cohort of 611 consecutive men with clinically localized (cT1-2, NX, M0) prostate cancer and no other treatment before documented progression. RESULTS: PFP for the entire study population was 78% at 5 and 76% at 10 years. The highest HR (0.09) was observed in the year immediately after surgery and dropped to 0 by year 7 (no patient recurred after year 6). Average annual HRs calculated for 3-year intervals resulted in steadily declining HRs over time for the entire study population and for all subsets, except those with a cancer pathologically confined to the prostate. Overall, the more ominous the prognostic factor, the higher the initial HR. For poorly differentiated cancers (biopsy Gleason sum 8 to 10), the HR was high in years 1 and 2 and dropped rapidly to 0 thereafter. CONCLUSIONS: Prostate-specific antigen (PSA) progression after RP usually occurred early (77% within the first 2 years) and was largely due to understaging. Late recurrences were rare in patients who were regularly evaluated with PSA. However, because the confidence intervals in our study were broad, larger patient populations with longer follow-up are needed for a definitive establishment of the time, course, and pattern of recurrence after surgery. PMID- 9218026 TI - Prognostic significance of detection of prostate-specific antigen transcripts in the peripheral blood of patients with metastatic androgen-independent prostatic carcinoma. AB - OBJECTIVES: To evaluate the prognostic significance of reverse transcriptase polymerase chain reaction (RT PCR) detection of prostate-specific antigen (PSA) mRNA in relation to survival in patients with metastatic androgen-independent prostatic carcinoma (AIPC). METHODS: Peripheral blood from 122 men (64 from Memorial Sloan-Kettering Cancer Center [MSKCC] and 58 from the Dana Farber Cancer Institute [DFCI]) with metastatic (Stage D2) AIPC was analyzed for PSA mRNA using RT PCR. Forty-one controls without prostatic carcinoma were also evaluated. RESULTS: RT PCR positivity for PSA mRNA was present in 24 of the 64 (38%) patients seen at MSKCC and in 26 of the 58 (45%) patients followed at DFCI. All control individuals were PSA PCR negative. There was a significant correlation between RT PCR positivity and decreased survival in each of the Memorial and Dana Farber population (P = 0.028 and 0.039, respectively). Serum PSA (at time of blood collection for PCR) was not predictive of survival as a continuous variable in the MSKCC [P = 0.31] and the DFCI (P = 0.09) groups. RT PCR for PSA mRNA was found to be independent from and superior to serum PSA in predicting survival in both the MSKCC and DFCI populations (P = 0.048 and P = 0.027, respectively). CONCLUSIONS: The detection of PSA mRNA in the peripheral blood by RT PCR is a predictor of survival in patients with metastatic AIPC, and PCR is superior to a single serum PSA measurement. Further studies are needed to test the value of this factor in comparison to and coupled with other prognostic parameters. PMID- 9218027 TI - Upsizing of the artificial urinary sphincter cuff to facilitate spontaneous voiding. AB - OBJECTIVES: Following placement of an artificial urinary sphincter (AUS) in the male child, functional and mechanical alterations can ensue, resulting in an inability to void spontaneously. One possible mechanical etiology in the patient entering puberty is prostatic growth within the fixed mechanical confines of the AUS cuff, resulting in progressive bladder outlet obstruction. Unrecognized infravesical obstruction can, in turn, lead to upper urinary tract deterioration, sepsis, or renal failure. We evaluated the effect of somatic growth and maturation of the male urethra on voiding dynamics in boys with an AUS to specifically determine whether revision of the sphincter cuff (ie, upsizing) is beneficial in restoring the ability to void spontaneously. METHODS: A retrospective review of 124 children with an AUS was performed. Eleven boys were identified whose bladder neck cuffs were later upsized in an attempt to improve bladder emptying. All boys were prepubertal at the time of original cuff placement. The average interval between the initial operation and cuff upsizing was 5 years. RESULTS: Following original sphincter placement, 8 patients emptied to completion spontaneously and 3 patients emptied by intermittent catheterization. All eight of the spontaneous voiders experienced progressive difficulty emptying after they entered puberty and ultimately had to rely on clean intermittent catheterization to empty completely. Follow-up subsequent to cuff exchange averaged more than 5 years (range 1 to 10 years). Despite an average increase of 10 mm in cuff size, all patients continued to depend on intermittent catheterization to empty completely. CONCLUSIONS: Upsizing the bladder cuff in the maturing male who experiences difficulty with bladder emptying does not restore the ability to void spontaneously. PMID- 9218028 TI - Use of a de-epithelialized local skin flap in hypospadias repairs accomplished by tubularization of the incised urethral plate. AB - OBJECTIVES: Snodgrass recently described a form of urethral tubularization with longitudinal incision of the urethral plate to create an elliptical meatus. To prevent fistulae, a transverse island of dorsal subcutaneous tissue was used to cover the repair. The generation of this flap may compromise blood supply to the skin that is used in the skin closure, and predispose to penile torsion. We have modified the technique to address these concerns, while accomplishing the major goal of functional success-particularly minimizing or eliminating fistulae. METHODS: Eighteen boys, aged 6 months to 6 years, with distal or midshaft hypospadias, underwent a one-stage repair using a modification of Snodgrass' technique. Rather than a transverse island flap of subcutaneous tissue, a local de-epithelialized skin flap was used to cover the urethroplasty. RESULTS: Sixteen patients have returned for follow-up, and all patients have an excellent cosmetic and functional result with an elliptical glanular meatus. There have been no cases of fistula or meatal stenosis. CONCLUSIONS: Distal hypospadias repair utilizing a de-epithelialized local skin flap to cover a tubularized incised urethral plate gives an excellent cosmetic and functional result. Optimal blood supply to the ultimate skin coverage is preserved, and penile torsion is avoided. PMID- 9218029 TI - Combined percutaneous antegrade and cystoscopic retrograde ureteral stent placement: an alternative technique in cases of ureteral discontinuity. AB - We describe an alternative method of double J stent placement for ureteral transection following the failure of traditional antegrade and retrograde approaches. Cystoscopically, a guidewire was placed across the distal ureteral segment and was advanced into a urinoma cavity at the level of the transected ureter. Subsequently, an antegrade approach was used to place a gooseneck snare through the proximal ureteral segment into the urinoma cavity. The guidewire was grasped with the snare and pulled through the percutaneous access site. A double J ureteral stent was then placed using the typical antegrade method. PMID- 9218030 TI - Ureteroscopic biopsy: technique and specimen preparation. AB - Because tissue samples obtained ureteroscopically are small, the techniques for biopsy and for handling and processing the samples are crucial. Our aim is to describe the biopsy technique in great detail so other centers can reproduce it. Patients were evaluated by retrograde ureteropyelography and ureteroscopy for diagnosis. The entire collecting system was examined using a combination of small diameter rigid and flexible ureteroscopes. Samples were retrieved by aspiration, saline lavage, or, when possible, biopsy of visible tumor by a basket or cup forceps. Multiple samples were obtained from all patients. Fresh specimens were hand delivered to the cytopathology laboratory, where they were evaluated with the cytospin technique. A cell block was prepared whenever there was any visible tissue in the sample. Since we have practiced this technique of handling specimens, our ability to diagnose and grade upper tract neoplasms ureteroscopically has improved markedly. Use of this technique can improve the diagnostic accuracy of ureteroscopic biopsy. PMID- 9218031 TI - Bilateral perinephric urinomas. PMID- 9218032 TI - Collagen mass in prostatic bed after radical retropubic prostatectomy. PMID- 9218033 TI - An uncommon urologic presentation of a supernumerary breast. AB - Polythelia and polymastia are common developmental abnormalities of the breast and nipple which usually present as small lesions along the mammary line, an embryologic line that extends bilaterally from the axillary regions to the inguinal ligaments. These lesions have been reported in various locations outside the mammary line. We report an unusual location of polymastia in the perineum of a newborn male. A brief discussion of the clinical relevance of such lesions is also included. PMID- 9218034 TI - Adrenal insufficiency from bilateral adrenal hemorrhage after total knee replacement surgery. AB - Adrenal hemorrhage is a rare cause of adrenal insufficiency in adults. We examine the incidence, etiology, diagnosis, and therapy of adrenal insufficiency secondary to adrenal hemorrhage. This case illustrates the nonspecific presentation of adrenal insufficiency and the necessity of maintaining a high index of suspicion in a clinically confusing scenario. PMID- 9218035 TI - Bilateral renal vein thrombosis in a newborn: a case of prenatal renal vein thrombosis. AB - Neonatal renal vein thrombosis (RVT) is a well described entity that typically presents after a variety of neonatal stresses. Prenatal RVT is a less common entity found incidentally on prenatal imaging. We report a case of neonatal RVT and a probable prenatal RVT. PMID- 9218036 TI - Nephron-sparing surgery in a renal allograft. AB - A 48-year-old woman underwent renal transplantation of an organ from a living related donor in 1978. She experienced excellent graft function for 18 years. Recent evaluation for hypertension revealed two large solid masses involving the allograft. Nephron-sparing excision of two renal cell carcinomas was performed with preservation of renal function. Genitourinary malignancies in transplant recipients and partial nephrectomy in renal allografts are reviewed. PMID- 9218037 TI - Obstructive anuria and dermatomyositis: a unique association. AB - This case is the first reported presentation of obstructive anuria due to extrinsic ureteral compression in a patient with dermatomyositis. Further evaluation revealed disseminated sigmoid colon adenocarcinoma. Conservative treatment of ureteral obstruction by internal double-J stents achieved appropriate palliation. PMID- 9218038 TI - A pilot study of energy utilization patterns during different transurethral electrosurgical treatments of the prostate. AB - OBJECTIVES: During a prospective randomized study of prostatic and periprostatic heating during transurethral electrosurgical treatment, energy utilization was studied with respect to electrode configuration and prostate size. METHODS: Patients were stratified for gland volume (transrectal ultrasound [TRUS] 50 cc or less and more than 50 cc) and randomized to treatment either with loop resection (transurethral resection of the prostate [TURP]) or electrovaporization (transurethral electrovaporization [TUEVAP]. VaporTrode-Grooved Bar, CIRCON ACMI). Power was provided by a radiofrequency unit (Force FX, Valleylab) initially set at 150 W. A passive feed-through system was connected to the patient circuit to record current and voltage at 10 Hz during each activation of the cut mode in real time. RESULTS: Patients (6 per group) were well matched for prostate volume (P < 0.57) and operating time (P < 0.33). Power settings were also similar (120 to 190 W). Both total energy utilization (P < 0.025) and energy used per minute of treatment (P < 0.004) were greater for TUEVAP than for TURP. The higher energy deposition per unit time for TUEVAP was not associated with undesirable periprostatic heating. For TURP, more energy was used per unit time for each gram resected in small prostates than in larger glands. Comparing energy consumption per minute per cubic centimeter of prostate, we found a 2:1 ratio between TUEVAP and TURP in large prostates, which increased to 3.4:1 (P < 0.049) in small glands. CONCLUSIONS: For the same panel power settings, more energy is deposited at the tissue interface during TUEVAP than during TURP. This extra energy provides better surface hemostasis without undesirable deep heating and can be explained by the larger contact surface and contact time (slower speed of excursion) of the VaporTrode than a regular loop. The novel observation that more energy is required for small prostates during both treatments suggests that these glands have different electrical properties and higher tissue impedance than larger glands. PMID- 9218040 TI - Human papillomavirus detection by polymerase chain reaction in benign and malignant prostate tissue is dependent on the primer set utilized. AB - OBJECTIVES: Prior investigations evaluating the presence of human papillomavirus (HPV) in prostatic tissue by polymerase chain reaction (PCR) technology have yielded detection rates of 0% to 100%. Contamination by viral DNA from prostatic urethral colonization or less than optimal laboratory conditions have been suggested to explain this discrepancy. In addition, the various investigations have differed in the specific oligonucleotide primers utilized for amplification and, therefore, have searched for different segments of the viral genome. The objective of this study is to address these differences. METHODS: Forty-one archival radical prostatectomy specimens were evaluated, identifying areas of normal and abnormal histology. Meticulous technique was used during tissue acquisition, histologic confirmation, DNA isolation, PCR amplification, polyacrylamide gel electrophoresis, and staining. Primers for a 126- and 99-base pair (bp) fragment of the E6 portion of HPV 16 as well as a consensus primer for the L1 portion of the papillomavirus genome were utilized. RESULTS: Of the normal prostatic tissues, 13.5% (5/37) contained the 126-bp E6 viral DNA as did 33.3% (7/21) of benign prostatic hyperplasia (BPH) samples, 25% (5/20) of dysplasia, 18.2% (2/11) of Gleason grades 1 and 2 adenocarcinoma, 25.9% (7/27) of Gleason grade 3 adenocarcinoma, and 6.7% (1/15) of Gleason grade 4 adenocarcinoma. Sections from the urethras of the prostatectomy specimens contained viral DNA in 31.7% (13/41). Viral detection was variable among different specimens in the same patient. With amplification for the 99-bp fragment of HPV 16, 1 of 37 normal (2.7%), 2 of 21 BPH (9.5%), 1 of 20 dysplasia (5.0%), and 2 of 53 cancer (3.8%) specimens revealed HPV DNA. In none of the specimens was DNA amplified using primers for a 450-bp fragment of the L1 portion of HPV. CONCLUSIONS: Previously published discrepancies in HPV detection may be solely due to the differences in primer sets utilized. PMID- 9218039 TI - Preservation of bladder tissue in different solutions for varying times. AB - OBJECTIVES: To evaluate three popular storage media and the effect of 24-hour cold storage on bladder tissue. METHODS: Guinea pig bladders were stored in three solutions: UW solution (a media used for transplant organs), Reznikoff solution [cell culture medium], and Krebs' solution with and without aeration. RESULTS: Cell potassium and sodium concentrations and total tissue water (a measurement of cell swelling) are important parameters for evaluating tissue damage. Reznikoff solution and Krebs' solution without gases maintained tissues for 24 hours with the least tissue damage; these solutions require no special equipment or attention. Twenty-four hour uniterrupted aeration of Krebs' solution caused the greatest degree of cell swelling with possible redistribution of receptors and required adjustment and regulation of the preservation apparatus. UW solution induced dehydration of cells, required the longest recovery period after cold storage, and is far more expensive than the other solutions. CONCLUSIONS: Reznikoff solution caused consistent relative changes in smooth muscle receptors and was superior to aerated Krebs' and UW solutions for 24-hour bladder tissue storage. It is unnecessary to aerate Krebs' solution during 24-hour cold storage. PMID- 9218041 TI - Hippocrates and urology: the first surgical subspecialty. AB - Hippocrates, who is generally considered a focal point for the start of western medical tradition, left behind a corpus of medical writings that constituted the first recorded comprehensive health system. The pivotal point of the Hippocratic corpus was the Hippocratic Oath, which outlined the duties of healers of his school, but demarcated lithotomy as a practice that was off limits to his fellow physicians. Surgery for bladder stone, urology in essence, was thus the first specifically identified surgical subspecialty. PMID- 9218042 TI - Reduction of paraphimosis with hyaluronidase. PMID- 9218043 TI - Balloon-wire retrieval of a migrated urethral stent in the early postoperative period. PMID- 9218044 TI - Circumcision in infancy. PMID- 9218045 TI - [The Bengmark principal. Placement and fixation of jejunal catheters in enteral feeding]. PMID- 9218046 TI - [Current knowledge on epilepsy, schizophrenia and spastic conditions. Neuro Forum, Dresden, 5 April 1997]. PMID- 9218047 TI - [Current knowledge on epilepsy, schizophrenia and spastic conditions. Neuro Forum, Dresden, 5 April 1997]. PMID- 9218048 TI - [Urinary incontinence--diagnosis, therapy, management]. PMID- 9218049 TI - [Management of seborrhoic dermatitis]. PMID- 9218050 TI - [Fungal infections of the skin. Itraconazol means a great advance in therapy]. PMID- 9218051 TI - [Rational step-by-step therapy of diabetic polyneuropathy]. PMID- 9218053 TI - [Atherosclerosis and nephropathy--calcium antagonists in preventive therapy]. PMID- 9218052 TI - [Lansoprazol: acid blocking in peptic disorders]. PMID- 9218054 TI - [HIV-related aspects in research and therapy]. PMID- 9218055 TI - [Value of acetylcholine in the management of Alzheimer disease]. PMID- 9218056 TI - [Drug therapy of panic disorders. Kava-specific extract WS 1490 compared to benzodiazepines]. PMID- 9218057 TI - [Therapy of the status epilepticus]. PMID- 9218058 TI - [Therapy of dementia]. PMID- 9218059 TI - [Special aspects of antidepressive therapy]. PMID- 9218060 TI - [Zotepin--its value and potentials for further development]. PMID- 9218061 TI - [From James Parkinson to modern Parkinson-therapy]. PMID- 9218062 TI - [Muscle pain--a multifaceted symptom]. PMID- 9218063 TI - [Enteral MRI contrast media--advances in magnetic resonance tomography of the abdomen and pelvis]. PMID- 9218064 TI - [10 years of AICD (Automated Implanted Cardiac Defibrillator) therapy in Germany]. PMID- 9218065 TI - Review article: the management of severe ulcerative colitis. AB - Severe ulcerative colitis is a potentially life-threatening condition but the mortality has fallen dramatically over the past 30-40 years. It is now less than 2%, including surgical mortality, and should only be seen in patients with significant co-existing disease. Early recognition of the severity of the colitis, intensive medical therapy, close liaison between physician and surgeon, and prompt surgery when necessary have all contributed to this improved outcome. Despite the use of high-dose intravenous corticosteroids, 20-30% of patients will make a poor response and will require urgent surgery. The use of intravenous cyclosporin has proved effective at reducing the immediate surgical rate in this group of unresponsive patients and appears safe. Whether cyclosporin reduces the need for surgery in the longer term is much less certain. Clinical, radiological, endoscopic and laboratory parameters can now be used to predict the course of a severe attack. These help in the timing of urgent surgery and are potentially helpful in determining when to begin other therapies such as cyclosporin. PMID- 9218066 TI - Review article: the therapy of gastrointestinal infections associated with the acquired immunodeficiency syndrome. AB - Although there have been dramatic strides in the therapy of human immunodeficiency virus infection over the last few years, the number of infected people world-wide is tremendous and, at least in developing countries, continues to expand. Complications which involve the gastrointestinal tract are common in these patients, because the gut is a major site for involvement by opportunistic infections and neoplasms in patients with the acquired immunodeficiency syndrome. It is important to recognize the clinical spectrum of gastrointestinal diseases, as well as the appropriate and most cost-effective diagnostic strategies, as therapies for a number of these disorders are both widely available and high effective. This review summarizes the major gastrointestinal infections which are seen in patients with the acquired immunodeficiency syndrome, and their treatment. PMID- 9218067 TI - Clinical economics review: liver transplantation. AB - The economic assessment of liver transplantation is complex and involves the cost of the transplant procedure, the cost of alternative treatments and the positive economic contribution of successful outcomes. The one-year survival rates are currently in the order of 80-90% for elective transplants and 60-75% for emergency transplantation. The average cost of a liver transplant operation in the UK is between pounds 32,000 and pounds 45,000. Factors which have a major influence on costs are aetiology, patient status at the time of transplantation, retransplantation and duration of hospital stay. Rejection episodes, renal impairment and surgical complications are of intermediate importance. The cost of transplantation must be balanced against the cost of alternative treatments that would be required if the patient were not transplanted and the positive economic benefit derived from excellent rehabilitation in the majority of recipients. PMID- 9218068 TI - Clinical economics review: nutritional support. AB - Nutritional support currently accounts for about 1% of the total health care costs in the USA. Interestingly, most of the prospective randomized controlled trials to date have not been able to demonstrate that this therapeutic intervention alters morbidity or mortality. In fact, parenteral nutritional support may predispose the recipients to developing systemic infections. There have been a few areas in which nutritional support may be of benefit. Enteral supplements given to underweight women who suffer hip fractures reduce the hospital stay and, presumably, overall cost. Preoperative parenteral nutritional support may produce a small absolute reduction in post-operative morbidity, but its cost becomes prohibitive. Preoperative enteral nutritional support, especially if carried out in the home, may be of benefit (using the most optimistic interpretation of a small number of trials); if so, it is an economically defensible intervention. Particular nutrients or diets may have specific effects on certain disease processes. Indirect comparisons have suggested that elemental diets can be used to treat flares of Crohn's disease (perhaps because putative food antigens are removed). However, corticosteroid therapy is more efficacious. Furthermore, it is less expensive to employ 6 mercaptopurine as the next modality in steroid failures. Branched-chain amino acid infusions may have some effect on hepatic encephalopathy, but again, lactulose is less expensive. Nutritional support is one area of medicine in which there has been far more enthusiasm than the data justify. Disease-associated malnutrition probably is a secondary phenomenon, not an important cause of morbidity. The widespread use of this modality cannot be justified in a cost constrained health care system. PMID- 9218069 TI - Prognostic factors influencing relapse of oesophagitis during maintenance therapy with antisecretory drugs: a meta-analysis of long-term omeprazole trials. AB - BACKGROUND: This meta-analysis investigated factors that may affect the risk of relapse of oesophagitis, and evaluated the predictive value of symptoms for the presence of relapse during long-term treatment. METHODS: Individual data from 1154 patients included in five independently conducted, randomized, long-term clinical trials of the efficacy of different dosage regimens of omeprazole, standard ranitidine treatment and placebo for the prevention of relapse of oesophagitis were pooled for this meta-analysis. The therapeutic regimens studied were omeprazole 20 mg o.m. (OME20) in 366 patients, omeprazole 10 mg o.m. (OME10) in 225 patients, omeprazole 20 mg weekends (OMEW) in 235 patients, ranitidine 150 mg b.d. (RAN) in 179 patients, or placebo (PLA) in 149 patients. RESULTS: OME20 maintained 82.4% (95% CI: 78.2-86.6%) of patients in endoscopic remission over the 6-month period compared to 71.9% (95% CI: 65.5-78.3%) for OME10, 52.3% (95% CI: 44.4-60.1%) for RAN, 42.7% (95% CI: 35.8-49.5%) for OMEW, and 10.6% (95% CI: 5.0-16.3%) for PLA. A Cox's regression analysis of time to recurrence of oesophagitis showed that four factors were associated with a higher relapse rate during placebo and active maintenance therapy: (a) pre-treatment severity of oesophagitis (P < 0.0001), (b) young age (P = 0.01), (c) non-smoking (P = 0.02) and (d) moderate/severe regurgitation before entry into the trials (P = 0.03). Asymptomatic relapse of oesophagitis was uncommon, being found in only 8.6% of the patients. CONCLUSIONS: Maintenance treatment with omeprazole 10 and 20 mg daily is superior to all other regimens tested, and is only marginally influenced by the pretreatment severity of oesophagitis. Age contributes to the factors that influence the outcome during long-term treatment with omeprazole. Symptom relief is highly predictive for the maintenance of healing. PMID- 9218070 TI - The effect of decaffeination of coffee on gastro-oesophageal reflux in patients with reflux disease. AB - BACKGROUND: Patients with reflux disease often complain of heartburn after ingestion of coffee. Induction of gastro-oesophageal reflux has been demonstrated by pH-metry following the intake of coffee in healthy volunteers. The reflux was reduced when the coffee had undergone a decaffeination process. The aim of this study was to investigate the effect of decaffeination of coffee on reflux in patients with reflux disease. METHODS: Seventeen reflux patients underwent two osesophageal 3-h pH measurements. The patients received, in a double-blind study design in a randomized order, 300 mL of either regular or decaffeinated coffee together with a standardized breakfast. The fraction time oesophageal pH < 4 was calculated during the three postprandial hours. RESULTS: For regular coffee the fraction time was calculated to a median of 17.9% with a range of 0.7-56.6%. The fraction time was significantly reduced to 3.1% (0-49.9%) after ingestion of decaffeinated coffee. CONCLUSION: The amount of gastro-oesophageal reflux induced by the intake of regular coffee in patients with reflux disease can be reduce by the decaffeination of coffee. PMID- 9218071 TI - Maintenance therapy with cisapride after healing of erosive oesophagitis: a double-blind placebo-controlled trial. AB - BACKGROUND: There are few data on the role of prokinetic agents as maintenance therapy in moderately severe reflux oesophagitis despite the high relapse rate of this condition after healing. AIMS: To determine whether cisapride is more effective than placebo as maintenance therapy after healing of moderate erosive oesophagitis in two respects: first, in preventing symptomatic relapse and preserving quality of life; and, second, in improving oesophageal motor function. PATIENTS: Forty-two patients whose grade II-III oesophagitis had been healed with omeprazole were randomized to receive either cisapride 20 mg nocte or placebo for 6 months. Oesophageal pH monitoring and manometry were performed before starting maintenance therapy and after 4 weeks, and symptomatic status and quality of life were assessed at weeks 0, 4, 13 and 26. RESULTS: After 4 weeks of maintenance therapy, lower oesophageal sphincter pressure improved in the cisapride group (16.4-21.9 mmHg, P = 0.01) but not in the placebo group (25.5-22.7 mmHg, P = 0.2). Oesophageal pH monitoring showed no significant changes in either group. Sixteen (76%) cisapride patients and 12 (57%) placebo patients withdrew within 4 weeks owing to symptomatic relapse (P = 0.2). After 26 weeks, 21 (100%) cisapride and 17 (81%) placebo patients had relapsed (log-rank analysis of survival time P = 0.07). Quality of life parameters deteriorated in both treatment groups to a similar degree. CONCLUSION: Maintenance therapy with cisapride 20 mg nocte improves the lower oesophageal sphincter pressure in patients whose oesophagitis has been healed with omeprazole. However, cisapride is no better than placebo in preventing symptomatic relapse or deterioration in quality of life. PMID- 9218072 TI - Comparison of two dosing regimens of cisapride in the treatment of reflux oesophagitis. AB - AIM: In an international, multicentre, double-blind trial, to document the therapeutic equivalence of two dosing regimens of cisapride on endoscopic healing and symptom improvement in patients with proven reflux oesophagitis grade I or II (Savary-Miller). METHODS: Four hundred and seven patients were randomly allocated to treatment with either cisapride 10 mg q.d.s. or cisapride 20 mg b.d. for 8-12 weeks depending on whether complete healing was found at endoscopy. The primary parameters of efficacy were cure of oesphagitis and improvement of the reflux symptom score. RESULTS: The healing rates at endpoint were 73% in both treatment groups. The mean total reflux symptom score decreased from baseline to endpoint from 7.9-2.1 (cisapride 10 mg q.d.s) and 7.9-2.5 (cisapride 20 mg b.d.). Each of the two treatment regimens was well tolerated. The most frequently (6.9%) reported adverse event (diarrhoea) was mild or moderate in most cases and can be explained by pharmacological action of cisapride. CONCLUSIONS: The results of the study demonstrate that cisapride 10 mg q.d.s. and 20 mg b.d. are equivalent in terms of efficacy and safety in the treatment of reflux oesophagitis. PMID- 9218073 TI - The long-term management of patients with bleeding duodenal ulcers. AB - BACKGROUND: Gastrointestinal haemorrhage is a common complication of duodenal ulcers. Patients who bleed are at substantial risk of recurrent bleeding. AIM: To determine whether appropriate therapeutic steps were taken to reduce the risk of recurrent haemorrhage in patients with a bleeding duodenal ulcer. METHODS: The management of patients surviving a duodenal ulcer bleed in the University Hospital. Nottingham, was assessed by case-note review before (1993) and after (1995-1996) institution of clinical guidelines. The following measures aimed at reducing the risk of recurrent haemorrhage were considered appropriate: stopping non-steroidal anti-inflammatory drugs (NSAIDs) when these were implicated in bleeding; successful eradication of Helicobacter pylori if present; and long-term maintenance acid-suppression therapy. RESULTS: In 1993, appropriate steps were taken to reduce the risk of recurrent haemorrhage in only 48% of cases. Following the institution of guidelines, management improved significantly in 1995-1996 (appropriate in 83% of cases, P < 0.001), was associated with increased referral to gastroenterologists (P < 0.001), improved patient compliance with follow-up (P < 0.05), and more rigorous attempts to identify (P < 0.001) and ensure clearance (P < 0.001) of H. pylori. CONCLUSION: In this study, inadequate long-term management of patients with a bleeding duodenal ulcer was common. This was to a failure to adopt strategies aimed at reducing the risk of ulcer relapse and rebleeding. The quality of care improved significantly following the institution of guidelines and encouragement to refer to gastroenterologists. PMID- 9218074 TI - Comparison of ranitidine and lansoprazole in short-term low-dose triple therapy for Helicobacter pylori infection. AB - AIM: To evaluate the efficacy and safety of two 1-week low-dose triple-therapy drug regimens involving antisecretory drugs for Helicobacter pylori infection. 99 patients with H. pylori infection were treated with either lansoprazole or ranitidine used together with clarithromycin and metronidazole. METHODS: The drug combination and administration periods in the proton pump inhibitor group were lansoprazole 30 mg o.m., clarithromycin 200 mg b.d. and metronidazole 250 mg b.d., all given for 7 days (LCM group). The ranitidine group received ranitidine 150 mg b.d., clarithromycin 200 mg b.d. and metronidazole 250 mg b.d. also for 7 days (RCM group). The presence or absence of H. pylori was determined from gastric biopsy specimens taken from both the antrum and the body, by smear, culture and tissue section (Giemsa stain). Cure was defined as failure to find evidence of H. pylori infection 4 weeks after antimicrobial therapy had ended. RESULTS: The cure of H. pylori infection was 88% in the LCM group (44 of 50; 95% confidence interval (CI) = 79-97%) and 92% in the RCM group (45 of 49; 95% CI = 84-99%). The incidence of adverse events was 16% and 18% for the two groups, respectively. CONCLUSIONS: No significant differences in cure rate and safety profiles were noted between the two regimens, suggesting that moderate acid inhibition using an H2-blocker is sufficient to achieve optimal H. pylori eradication. PMID- 9218075 TI - Doubling the omeprazole dose (40 mg b.d. vs. 20 mg b.d.) in dual therapy with amoxycillin increases the cure rate of Helicobacter pylori infection in duodenal ulcer patients. AB - BACKGROUND: Several studies have shown that dual therapy with omeprazole and amoxycillin may cure Helicobacter pylori infection. However, the optimum dose of omeprazole has still to be established. METHODS: An international, randomized, double-blind multicentre trial was conducted in patients with duodenal ulcers to compare the H. pylori cure rates obtained by dual therapy consisting of either omeprazole 20 mg b.d. plus amoxycillin 750 mg b.d. or omeprazole 40 mg b.d. plus amoxycillin 750 mg b.d. for 2 weeks. Dual therapy was followed by omeprazole 20 mg once daily for 2 weeks. Before entering the trial and 4 weeks after cessation of treatment H. pylori infection was assessed by histology and a 13C-urea breath test. RESULTS: 381 patients were randomized into the study, of whom 345 were evaluable for the all-patients-treated analysis of efficacy and 378 were valid for the evaluation of safety. Histology results showed that H. pylori infection was cured in 64 out 174 patients treated with omeprazole 20 mg b.d. plus amoxycillin and in 102 out of 171 patients treated with omeprazole 40 mg b.d. plus amoxycillin (37% vs. 60%; P < 0.001). Both treatment regimens were well tolerated, with adverse events reported by 29 (15.2%) and 35 patients (18.7%), respectively. CONCLUSIONS: This study has shown that dual therapy with amoxycillin 750 mg b.d. and omeprazole 40 mg b.d. is superior to dual therapy with amoxycillin and omeprazole 20 mg b.d. in patients with H. pylori-positive duodenal ulcers. Thus, a true dose-response relationship exists between omeprazole and treatment success. However, a combination of omeprazole with two of amoxycillin, clarithromycin and nitroimidazole is a preferable alternative for routine clinical use. PMID- 9218076 TI - Successful low-dose amoxycillin, metronidazole and omeprazole combination therapy in a population with a high frequency of metronidazole-resistant Helicobacter pylori. AB - AIM: Effective anti-Helicobacter pylori therapies with few side-effects are needed. We studied the effectiveness of a low-dose combination of metronidazole, amoxycillin and omeprazole for treatment of ulcer patients in Seoul, Korea. METHODS: Patients with gastric or duodenal ulcer received metronidazole (125 mg b.d.), amoxycillin (500 mg b.d.) and omeprazole (20 mg at bedtime) for 2 weeks. Endoscopic examinations were performed before treatment and at least 6 weeks after completion of antimicrobial therapy. H. pylori status was confirmed by histological examination of two gastric biopsies using the Genta stain. RESULTS: Seventy-nine patients (64 men, 15 women, mean age 46 years) with peptic ulcer were enrolled. H. pylori infection was cured in 56 (71%) 95% CI: 60-81%). The cure rate in non-smokers was significantly higher than in smokers (88% vs. 65%, P = 0.035). Twelve pre-treatment isolates were available and metronidazole resistance was noted in all; H. pylori infection was cured in 10. Thirty-six patients cured of H. pylori have been followed for 1 year (mean of 361 days) and 2 cases became reinfected (5.5%, 95% CI: 1-18%). CONCLUSIONS: The low-dose combination of metronidazole, amoxycillin and omeprazole was effective even the in face of metronidazole resistance. Recurrence of H. pylori infection is infrequent even in countries with a high prevalence of H. pylori infection. PMID- 9218077 TI - Furazolidone, amoxycillin, bismuth triple therapy for Helicobacter pylori infection. AB - BACKGROUND: Metronidazole-resistant Helicobacter pylori are generally the rule in developing countries such as Colombia. Developing countries need an effective, simple and inexpensive non-metronidazole therapy for H. pylori infection. AIM: To evaluate the combination of bismuth, furazolidone and amoxycillin for the treatment of H. pylori infection in Colombia. METHODS: Thirty patients with histologically documented H. pylori infection received the combination of bismuth subcitrate 240 mg b.d., furzolidone 100 mg q.d.s. and amoxycillin 500 mg q.d.s. for 14 days. Four or more weeks after ending therapy patients were re-endoscoped and gastric biopsies were obtained and examined using the Genta stain. Each slide was scored for presence, absence and density of H. pylori, active and chronic inflammation, intestinal metaplasia, erosions and atrophy. Cure was defined as the absence of H. pylori. RESULTS: All patients completed the course of therapy. Twenty-five patients were cured (86%, 95% CI: 65-94%). Mild, well-tolerated side effects were reported by six patients (20%). CONCLUSIONS: This combination of bismuth, furazolidone and amoxycillin fulfills the criteria for successful H. pylori therapy and appears particularly well suited for developing countries since it is simple, inexpensive and effective. Furazolidone-containing therapies may become especially useful in the face of a world-wide increase in H. pylori resistance to metronidazole and macrolides. PMID- 9218078 TI - Low cure rate of Helicobacter pylori infection with omeprazole and furazolidone dual therapy for one week. AB - BACKGROUND: Furazolidone is an inexpensive antibiotic that has considerable anti Helicobacter pylori activity in vitro. METHODS: Twenty-three patients with culture-proven H. pylori infection were treated for one week with a dual therapy containing omeprazole and furazolidone. RESULTS: Eradication succeeded in 10 of the first 20 evaluable patients (50%; 95% CI: 27.2-72.8%). This percentage was regarded as too low, and the study was terminated. Side-effects were mild. CONCLUSION: With the possible increase in resistance to metronidazole and clarithromycin world-wide, furazolidone may be useful alternative in the treatment of H. pylori infection. Dual therapy for one week, however, is not sufficient. PMID- 9218079 TI - Helicobacter pylori eradication using a 7-day regimen of low-dose clarithromycin, lansoprazole and amoxycillin. AB - AIM: To evaluate the efficacy of a 7-day regimen of clarithromycin 250 mg b.d., amoxycillin 1 g b.d., and lansoprazole 30 mg b.d. as a treatment for Helicobacter pylori infection. METHODS: H. pylori status of dyspeptic patients was assessed by 13C-urea breath test and at endoscopy by histology, culture and rapid urease testing of gastric biopsies. Fifty-one H. pylori-positive patients were treated with the above regimen. H. pylori status was reassessed by 13C-urea breath test not less than 28 days after completing treatment. Adverse events and compliance were evaluated. RESULTS: On an intention-to-treat basis. H. pylori infection was cured in 77% (95% CI: 65-88%) of patients. Minor side-effects including diarrhoea, nausea and taste disturbance were reported by 64% of patients. Ninety five per cent of patients consumed > 95% of tablets. Metronidazole resistance was 29% but all cultures were sensitive to amoxycillin and clarithromycin. CONCLUSION: This 7-day treatment with low-dose clarithromycin was moderately effective in curing H. pylori infection. Although compliance was excellent, there was a high frequency of minor adverse events. PMID- 9218080 TI - Lansoprazole 30 mg daily versus ranitidine 150 mg b.d. in the treatment of acid related dyspepsia in general practice. AB - AIM: To compare lansoprazole 30 mg daily with ranitidine 150 mg b.d. in the treatment of acid-related dyspepsia in general practice. METHODS: In a double blind, parallel group, randomized, mutlicentre study conducted in 32 general practices in the UK, 213 patients were randomized to receive lansoprazole 30 mg daily, and 219 to receive ranitidine 150 mg b.d., for 4 weeks. All patients had experienced symptoms of reflux-like or ulcer-like dyspepsia on at least 4 of the 7 days prior to the study; 75% had experienced dyspepsia in the past, and 74 of the lansoprazole patients and 77 of the ranitidine patients had documented histories of acid-related disorders, investigating by either radiology or endoscopy. RESULTS: After 2 weeks 55% of the lansoprazole patients and 33% of the ranitidine group were symptom-free (P = 0.001, chi 2 = 7.12) with corresponding 4 week figures of 69% and 44%, respectively (P = 0.001, chi 2 = 18.03). Similar figures were found at both 2 and 4 weeks for daytime and night-time heartburn and epigastric pain scores; in the lansoprazole group, at 4 weeks, 80% of patients were free of daytime heartburn and 81% of night-time epigastric pain, compared with 55% (P = 0.001, chi 2 = 15.44) and 65% (P = 0.01, chi 2 = 6.10) in the ranitidine group. CONCLUSION: Superior symptom relief for patients presenting with ulcer-like and reflux-like symptoms in general practice is provided by lansoprazole 30 mg daily compared with ranitidine 150 mg twice daily. PMID- 9218081 TI - Evaluation of a new quality of life questionnaire for patients with irritable bowel syndrome. AB - BACKGROUND: We describe the development and evaluation of a new disease-specific instrument, the Irritable Bowel Syndrome Quality of Life Questionnaire (IBSQOL), which was designed for use in patients with irritable bowel syndrome. The IBSQOL measures 10 domains found to be relevant to patients with irritable bowel syndrome: emotional health, mental health, health belief, sleep, energy, physical functioning, diet, social role, physical role, and sexual relations. METHODS: During its development and evaluation, the IBSQOL was administered to over 500 patients with irritable bowel syndrome--two groups of patients from tertiary care centres, three focus groups of 8-12 patients each, and 287 patients in a national irritable bowel syndrome support network. As a control, the IBSQOL was also administered to 37 patients who did not have irritable bowel syndrome but had other gastrointestinal disorders. Statistical analyses to test the reliability and validity of the IBSQOL were performed using Cronbach's alpha coefficient. RESULTS: Responses from the focus groups indicated that the IBSQOL was easy to complete and did not require too much time to fill out (approximately equal to 25 min). Statistical analyses of the final 30-item version of the IBSQOL demonstrated that it had both adequate validity and reliability (alpha > or = 0.60). A comparison of mean IBSQOL scores of persons with and without irritable bowel syndrome (but with other gastrointestinal conditions) showed no difference between the two groups with irritable bowel syndrome; however, scores for both irritable bowel syndrome groups were considerably lower than for the non irritable bowel syndrome group, suggesting better health-related quality of life in patients who do not have irritable bowel syndrome. This further demonstrated the validity of the IBSQOL in targeting questions and domains specific to patients with irritable bowel syndrome. CONCLUSIONS: Evaluation of the IBSQOL included testing the questionnaire in a large number of patients, which resulted in a revised and well-constructed instrument that demonstrated both adequate validity and reliability. The IBSQOL is currently being used in large-scale clinical trials to measure changes in quality of life in patients with irritable bowel syndrome following treatment intervention. PMID- 9218082 TI - Patient-perceived severity of irritable bowel syndrome in relation to symptoms, health resource utilization and quality of life. AB - AIM: In this study of patients with irritable bowel syndrome (IBS), we evaluated the relationship between patient-rated severity of IBS and patients' physical and psychological symptoms, health care resource use and quality of life. METHODS: One hundred and twenty-six patients diagnosed with IBS were administered a series of questionnaires, including the Bowel Symptom Checklist, the Symptom Checklist 90-R (a psychological symptom checklist), the IBSQOL (a disease-specific quality of life instrument), the SF-36 (a general health status instrument), and a health resource utilization assessment that measured health care use, time loss from work, impact on productivity, and days worked with symptoms. RESULTS: No relationship was found between IBS severity and gastrointestinal symptoms, except for a feeling of unpassed stool. IBS severity was also not related to psychological symptom severity. Direct traditional indicators of resource use (e.g. physician visits, hospital admissions and emergency room visits) were not significantly associated by severity level; however, indirect measures of resource use (e.g. number of days with pain, productivity and number of bed days) were related to severity. Quality of life was clearly associated with perceived IBS severity. Patients who rated themselves as very severe reported the lowest scores and had the poorest health for all quality of life dimensions measured. CONCLUSIONS: These findings suggest that perceived IBS severity is defined by the limitations the disease imposes, rather that by the symptoms. Patients with reduced productivity and decreased functioning for most of the quality of life indicators were those who rated their IBS as very severe. PMID- 9218083 TI - Zamifenacin (UK-76, 654) a potent gut M3 selective muscarinic antagonist, reduces colonic motor activity in patients with irritable bowel syndrome. AB - BACKGROUND: Zamifenacin is a new potent gut M3 selective muscarinic antagonist developed for possible use in irritable bowel syndrome. METHODS: In this multicentre, double-blind, parallel group, placebo-controlled study, the effect of a single dose of zamifenacin 10 mg or 40 mg on both fasting (30 min) and fed (60 min) colonic motor activity was assessed in 36 patients with irritable bowel syndrome (aged 25-68 years; 19 male). Colonic motility was recorded using a five channel solid-state catheter introduced by colonoscopy to a depth of 35 cm in an unprepared colon. RESULTS: Zamifenacin 40 mg profoundly reduced colonic motility, particularly after the meal (P < 0.05). This was reflected by a significant reduction in the mean amplitude of contractions, number of contractions, percentage duration of contractions, activity index and the motility index (P < 0.05). A smaller reduction in all the motility parameters was obtained with 10 mg zamifenacin, but these changes were not statistically significant. Three patients each on placebo and zamifenacin reported side-effects, but these were mild and transient. CONCLUSION: A single 40 mg dose of zamifenacin significantly reduces colonic motility in irritable bowel syndrome patients without significant antimuscarinic effects. The results of this study confirm that the concept of developing selective antimuscarinic agents may be a promising approach to the treatment of irritable bowel syndrome. Not only would such compounds benefit from not having the usual side-effects of anticholinergics but they might also offer much more in the way of dose flexibility. PMID- 9218084 TI - The absorption of low-dose methotrexate in patients with inflammatory bowel disease. AB - BACKGROUND: Recent clinical trials have demonstrated that methotrexate may have an important therapeutic role in the treatment of patients with inflammatory bowel disease, who are either refractory or intolerant to traditional medical therapy. The aim of this study was to evaluate the pharmacokinetics of low-dose oral methotrexate in patients with inflammatory bowel disease. METHODS: Methotrexate (12.5 mg) was given orally to nine patients with inflammatory bowel disease: five with Crohn's disease, and four with ulcerative colitis, and to six patients with rheumatoid arthritis who served as a control group. Blood samples were drawn at specific intervals to evaluate methotrexate plasma levels. RESULTS: Methotrexate was rapidly absorbed in all patients. Peak concentrations (Cmax) varied considerably, ranging from 0.25-0.87 micro M. The mean Cmax values were similar in all patient groups (0.59 +/- 0.12, 0.69 +/- 0.16 and 0.54 +/- 0.18 micro M, P not significant) for Crohn's disease, ulcerative colitis and rheumatoid arthritis, respectively. The mean area under curve in 120 min (AUC0 120) was also similar in all patient groups (32.9 + 11.3, 43.6 + 9.9 and 41.8 + 14.9 ng.min/mL, P not significant) for Crohn's disease, ulcerative colitis and rheumatoid arthritis, respectively. The mean time to reach Cmax, (tmax), varied between patient groups (84, 112 and 95 min, respectively, with a significant difference, P < 0.02, between the Crohn's disease and ulcerative colitis groups. A negative correlation was found between methotrexate dosage/kg and Cmax (r = 0.74) only in Crohn's disease patients but not in the other patient groups. CONCLUSIONS: Orally administered methotrexate is well absorbed in patients with inflammatory bowel disease including those with severe small bowel disease or resection. If methotrexate is proven to be effective in inflammatory bowel disease, it should be administered orally. PMID- 9218085 TI - Effect of acute pentoxifylline treatment in an experimental model of colitis. AB - BACKGROUND: The effect of acute pentoxifylline treatment in an experimental model of colitis was assessed using the trinitrobenzene sulphonic acid (TNBS)-induced rat model of colitis. METHODS: Animals were treated with intracolonic injection (250 microliters) of TNBS (50 mg in 50% ethanol) to induce inflammation and ulcers. Animals received pentoxifilline (100 mg/kg intracolonically) or saline 24 and 48 h following TNBS treatment. Five days following TNBS treatment, colons were dissected and scored according to the morphology damage score. The colons were then rolled longitudinally, fixed in formalin and embedded in paraffin. The collagen content of colonic sections were determined by a Sirius red-Fast green technique. RESULTS: Animals treated with TNBS alone had significantly higher gross morphology damage scores compared to animals treated with saline. Pentoxifylline significantly reduced the gross morphology damage score in animals receiving TNBS. Colonic collagen levels were significantly elevated in TNBS treated animals compared to animals receiving saline. Pentoxifylline treatment did not alter the collagen content of colons from TNBS-treated animals. CONCLUSION: TNBS treatment significantly elevates morphology damage score compared to controls. The results also suggest that colonic collagen was significantly elevated in animals treated with TNBS compared to controls. Pentoxifylline treatment was not sufficient to reduce the elevation in colonic collagen, although pentoxifylline treatment was sufficient to reduce the pathological changes due to TNBS, thus bringing the morphology damage score down to control levels. PMID- 9218086 TI - Studies of water and electrolyte movement for oral rehydration solutions (rice- and glucose-based) across a normal and secreting gut using a dual isotope tracer technique in a rat perfusion model. AB - AIMS: To establish a model to measure bidirectional flow of water from a glucose oral rehydration to solution (G-ORS) and a newly developed rice-based oral rehydration solution (R-ORS) using a dual isotope tracer technique in a rat perfusion model. To measure net water, sodium and potassium absorption from the ORS. METHOD: In vivo steady-state perfusion studies were carried out in normal and secreting (induced by cholera toxin) rat small intestine (n = 11 in each group). To determine bidirectional flow of water from the ORS the animals were initially with tritium, and deuterium was added to the perfusion solution. Sequential perfusate and blood samples were collected after attainment of steady state conditions and analysed for water and electrolyte content. RESULTS: There was significant increase in net water absorption from the R-ORS compared to the G ORS in both the normal (P < 0.02) and secreting intestine (P < 0.05). Water efflux was significantly reduced in the R-ORS group compared to the G-ORS group in both the normal (P < 0.01) and the secreting intestine (P < 0.01). There was an increase in sodium absorption in the R-ORS group compared to the G-ORS. The G ORS produced a significantly greater blood glucose level at 75 min compared to the R-ORS (P < 0.03) in the secreting intestine. CONCLUSIONS: This study demonstrates the improved water absorption from a rice-based ORS in both the normal and secreting intestine. Evidence that the absorption of water may be influenced by the osmolality of the ORS was also demonstrated. PMID- 9218087 TI - Transdermal delivery of erythromycin lactobionate--implications for the therapy of gastroparesis. AB - BACKGROUND: The treatment of many diseases may be complicated by abnormalities in gastric emptying. Gastric motor dysfunction may lead to unpredictable food and medication delivery to the small intestine, their site of absorption. Prokinetic agents improve gastric motility, but orally administered drugs are unreliably absorbed, thereby limiting their effectiveness. A method of delivering prokinetic agents which bypasses the gastrointestinal tract could lead to more effective treatment. METHODS: Skin samples from rat, hairless mouse and man were placed in an in vitro diffusion chamber. The epidermal side of the skin was exposed to erythromycin lactobionate and passage of the drug across the skin sample monitored and quantitated by high-performance liquid chromatography with UV detection. RESULTS: Erythromycin passes across all skin types tested. Steady state flux across hairless mouse skin was greater than for rat, full thickness human skin and human epidermis. In the first 3 h following introduction of erythromycin lactobionate, 1.85 mg/cm2 crossed human epidermis. Given that a dose of 50 mg may exert prokinetic effects in vivo in man, increasing the patch size to approximately equal to 28 cm2 should provide therapeutic levels of drug within 3 h. CONCLUSIONS: Erythromycin lactobionate, when administered transdermally, can be delivered at levels sufficient to treat gastroparesis. This technique warrants in vivo investigation. PMID- 9218088 TI - Gastric emptying of semisolid meal unaltered by 3 days' administration of a sustained-release preparation of the nitric oxide donor, isosorbide dinitrate. AB - BACKGROUND: Evidence has accumulated that nitric oxide is involved in the regulation of gastrointestinal motor activity. We investigated whether nitric oxide derived from a sustained-release isosorbide dinitrate (Cedocard retard) had an effect on gastric emptying and on subjective feelings. METHODS: Twelve healthy males aged 23-32 years received at weekly intervals, for 3 days twice daily, either 20 mg isosorbide dinitrate, 40 mg isosorbide dinitrate, or placebo, under random double-blind conditions. After a further dose on day 4, subjects ate a 1168 kJ semisolid meal, the emptying of which was recorded scintigraphically for 50 min. RESULTS: Neither dosage of isosorbide dinitrate had an effect on emptying which differed from the effect of placebo and the effects of the two dosages were the same. The radioactivity remaining in the stomach 50 min postprandially was 68.5% +/- 4.5 S.E.M. after placebo, 65.4 +/- 5.6% after 20 mg isosorbide dinitrate and 66.1 +/- 4.4% after 40 mg isosorbide dinitrate. With 40 mg isosorbide dinitrate, all 12 subjects complained of persistent headache, whereas only slight headache was reported by 7 subjects on 20 mg isosorbide dinitrate and by 1 subject on placebo. CONCLUSION: Twenty and 40 mg doses of sustained-release isosorbide dinitrate twice daily had no effect on the gastric emptying of a semisolid meal, but dose-dependently induced headaches. PMID- 9218089 TI - Cisapride and erythromycin prokinetic effects in gastroparesis due to type 1 (insulin-dependent) diabetes mellitus. AB - BACKGROUND: Erythromycin, a macrolide antibiotic, has been shown to have gastric prokinetic effects and has been proposed as an alternative therapeutic option for diabetic gastroparesis. However, its efficacy has not yet been compared with that of other prokinetic drugs. AIMS: The purpose of the present study was to compare the effects of erythromycin (250 mg 60 min before meals) and cisapride (10 mg 30 min before meals) on gastric emptying of healthy subjects and insulin-dependent diabetics. PATIENTS AND METHODS: Six type 1 diabetic patients with a previous scintigraphic demonstration of gastroparesis and five healthy subjects were recruited for the study. Gastric emptying was scintigraphically studied by labelling the solid component of a standard test meal. Three scintigraphic studies, spaced at least 3 days apart, were carried out on each subject, basally and after erythromycin or cisapride. RESULTS: Cisapride significantly accelerated gastric emptying in both the healthy subjects and the diabetic patients without any significant effect on the lag-time, whereas erythromycin in addition to a significant improvement of the overall gastric emptying also showed a pronounced effect on the lag-time in both groups (controls 25 +/- 5 vs. 37 +/- 8 min, P < or = 0.04; diabetics 65 +/- 11 vs. 112 +/- 16 min, P < 0.03). CONCLUSIONS: Erythromycin may represent an effective therapeutic alternative to more established forms of treatment in patients with diabetic gastroparesis, especially when other drugs have failed. PMID- 9218090 TI - Cytoprotective effect of bismuth subsalicylate in indomethacin-treated rats is associated with enhanced mucus bismuth concentration. AB - BACKGROUND: Bismuth compounds prevent gastric injury from the short-term administration of nonsteroidal anti-inflammatory drugs. We studied the mechanisms underlying the gastroprotective actions of bismuth subsalicylate against indomethacin-induced injury in rats. METHODS: An in vivo microscopic technique was used in which acid output, surface cell intracellular pH (pHi), gastric mucus gell thickness and mucosal blood flow were measured simultaneously. Concentrations of bismuth in mucus were measured by atomic absorption. RESULTS: Indomethacin (60 mg/kg) significantly thinned the mucus gel layer and augmented the decrease of pHi during luminal acid superfusion, consistent with a weakened gastric mucosal barrier to acid. Bismuth subsalicylate partially reversed this effect of indomethacin on pHi, consistent with gastroprotection. Neither a prostaglandin-inhibiting but non-injurious dose of indomethacin (5 mg/kg), bismuth subsalicylate, or their combination affected mucus gel thickness or pHi homeostasis. In separate experiments, indomethacin (60 mg/kg) significantly increased gastric mucus bismuth concentration in rats given bismuth subsalicylate. CONCLUSION: Bismuth accumulation in the gastric mucus during the evolution of mucosal injury may play an important role in the gastroprotective effect of bismuth subsalicylate against indomethacin injury. PMID- 9218091 TI - Suppressive effect of tetraprenylacetone on gastric atrophy induced by short-term administration of N-methyl-N'-nitro-N-nitrosoguanidine in rats. AB - BACKGROUND: Several studies have been reported on the effects of various therapeutic agents in enhancing or suppressing the carcinogenic activity of N methyl-N'-nitro-N-nitrosoguanidine (MNNG). However, it is still unknown whether a mucosal protective agent could suppress its carcinogenic activity. METHODS: Twenty-five Wistar male rats were divided into four groups: group 1, MNNG alone; group 2, MNNG + tetraprenylacetone; group 3, control; group 4, tetraprenlacetone alone. MNNG 100 mg/mL, was freely given to groups 1 and 2, and tetraprenylacetone (200 mg/kg intraperitoneal) was additionally administered every other day to the rats in groups 2 and 4. The animals were sacrificed at 10 weeks and the gastric mucosa examined. RESULTS: Atrophic changes were observed in the antrum after 8 weeks of oral administration of MNNG. Furthermore, using immunohistological analysis with 5-bromo-2'-deoxyuridine (BrdU), the proliferative zone was found to be enlarged and shifted upward, although the BrdU labelling index of the proliferative zone was unaltered. Intraperitoneal administration of tetraprenylacetone every other day suppressed the MNNG-induced atrophic change and the alterations proliferative markers. Tetraprenylacetone alone did not have an effect either on morphological or proliferative markers. CONCLUSION: These observations suggest that gastric mucosal defensive factors may play critical roles in suppressing atrophic change inducing carcinogenesis by an exogenic carcinogen. PMID- 9218093 TI - Role of vasopressin in long-term preferences of sucrose and polycose under ad libitum and food-restricted conditions. AB - Vasopressin-deficient (DI) and vasopressin-containing (LE) rats were given continuous access to 32% sucrose and 32% polycose solutions under ad-libitum and food-restricted conditions in a long-term preference test. Although all animals preferred Polycose to sucrose in both conditions, food restriction introduced a stress that significantly increased the consumption of Polycose in DI rats. Considering total caloric intake, Polycose was preferred by both strains only under food restriction and the effect was greatly exacerbated in DI animals compared to LE animals. The differences observed between DI and LE animals in response to ad-libitum feeding and food-restriction stress indicate that vasopressin, directly and/or indirectly, influences the physical and metabolic functions and processes of the animals, which, in turn, affect their intake of Polycose. PMID- 9218092 TI - NSAIDs upregulate beta 2-integrin expression on human neutrophils through a calcium-dependent pathway. AB - BACKGROUND: Margination of circulating neutrophils (PMN) into the gastric microcirculation is an early and critical event in the pathogenesis of non steroidal antinflammatory drug (NSAID)-induced gastropathy. This effect is mediated through the upregulation of beta 2 integrins on the PMN surface. AIMS: To investigate whether indomethacin modulates: (1) Mac-1 expression; (2) Ca2+ mobilization ([Ca2+]i), protein kinase C and nitric oxide accumulation; and (3) mitogen-associated protein kinase phosphorylation in human PMN. METHODS: Human PMN were isolated by centrifugation through a double Ficoll gradient. [Ca2+]i was measured in PMN loaded with fura-2 and Mac-1 expression by flow cytometry. RESULTS: Indomethacin caused a concentration- and time-dependent upregulation of CD11b and CD18 expression and PMN adhesion to endothelial cells. Maximal upregulation of Mac-1 expression (40-50%) occurred after a 30-min incubation with 0.1mM indomethacin. The effect was prevented by removing the Ca2+. Ionomycin and thapsigargin caused a 7-10-fold increase in [Ca2+]i and a 2-4-fold increase in Mac-1 expression. Indomethacin induced a concentration-dependent phosphorylation of a 41-kDa mitogen-associated protein kinase. Tyrosine kinase inhibitors prevented the effect of indomethacin on Mac-1 expression and Ca2+ mobilization. Indomethacin and ionomycin increased superoxide generation, myeloperoxidase secretion and PMN adherence to endothelial cells and stimulated nitric oxide production. Indomethacin-induced Mac-1 upregulation was prevented by a nitric oxide synthase inhibitor. CONCLUSIONS: Indomethacin-induced upregulation of Mac-1 is mediated by changes in [Ca2+]i and nitric oxide. Phosphorylation of the 41-kDa mitogen-associated protein isoform is a previously unreported target of NSAID action. These effects might help to explain the ability of indomethacin to cause gastric neutrophil margination. PMID- 9218094 TI - Geometric analysis of macronutrient selection in the rat. AB - A conceptual framework is introduced which has been derived from work on insects. The scheme is intended to integrate studies of diet selection, regulation of amounts eaten, nutrient utilization, body composition and animal performance. Aspects of framework are illustrated with published data on macronutrient selection in the rat. An animal is viewed as moving through a multidimensional nutrient space, which is bounded by axes representing each required nutrient and within which lie optimal points of intake and nutrient allocation ("targets"). The aim is first to estimate the location of these functional optima experimentally, and then to interpret the responses of animals which are constrained from reaching them ("decisions of best compromise"). The framework can then be used to interpret data from animals reared under differing environmental conditions and to compare animals of differing developmental stage, genotype or nutritional state. PMID- 9218095 TI - Effects of deprivation level on humans' self-control for food reinforcers. AB - Deprivation level was manipulated in fourteen food- and water-deprived adult human females to examine its effects on self-control for food (choice of larger, more delayed access to apple juice over smaller, less delayed access to apple juice). Each subject was exposed to two treatments: (1) Consumption of a 500 g tomato soup preload just prior to self-control testing and (2) no soup preload. When subjects had consumed soup, they reported significantly less hunger and showed significantly more self-control as compared to when not having consumed soup. Additionally, when subjects had consumed soup, self-control decreased as a function of session time. Subjects who reported that they were currently dieting drank significantly less juice when they had previously consumed soup than when they do not previously consumed soup. Together, the results indicate that when subjects are more deprived they may be less able to wait for food reinforcers (i.e., show less self-control). Such behaviour may be adaptive in situations in which energy is needed to survive periods of food scarcity. PMID- 9218096 TI - Increasing willingness to taste novel foods: effects of nutrition and taste information. AB - This study was designed to examine the effects of various kinds of information on willingness to ingest novel foods in individuals varying in the extent to which they reported that nutritional concerns affected their food choices. Male and female volunteers ranging in age from 10 to 79 (N = 401), saw six familiar and six novel foods, and received no information, taste likability information, general nutrition information, or specific nutrition information about the whole set of foods. They rated their willingness to taste each food, with the clear implication that their willingness ratings would determine which foods they would taste later in the study. On a separate questionnaire, they also rated the factors influencing their food everyday choices, and these ratings were used to compute an "importance of nutrition" score for each individual. Results indicated that older subjects were generally more willing to try novel foods than younger ones, that general nutrition information was effective for high school and college students, and that specific nutrition information was influential for young adults. It was also found that general nutrition information increased willingness to taste novel food in subjects for whom nutrition is important and decreased such willingness in subjects for whom nutrition is not important. PMID- 9218097 TI - Temperament and food neophobia in children and their mothers. AB - We obtained measures of behavioral neophobia, rated neophobia, temperament, and liking for novel and familiar foods for 81 pairs of siblings (ranging in age from 5-11 years) and their mothers. The children's data revealed a decrease with age in behavioral but not rated neophobia, and there were positive correlations between both measures of neophobia and the temperament dimensions of emotionality, shyness, and negative reactions to foods. In addition, liking for good-tasting novel foods was negatively related to shyness and to negative reactions to foods. For mothers, both measures of neophobia were related to negative reactions to foods and unrelated to any other temperament dimensions. The data revealed little evidence of family resemblance (mother-child or sibling sibling) in food neophobia. PMID- 9218098 TI - Perceptions of starchy food dishes: application of the Repertory Grid Method. AB - The Repertory Grid Method (RGM) was applied to obtain an understanding of the characteristics used by U.K. consumers in discriminating amongst different common starchy food dishes, including potatoes, rice and pasta. Twenty-nine subjects generated a large number of constructs, relating to perceived nutrition, health physiological effect, sensory, and use attributes of these products. Coupling of RGM with Generalized Procrustes Analysis produced detailed qualitative and quantitative information describing common and individual characteristics of particular dishes. The results indicate that starchy foods are in general seen as "filling", but specific products are clearly discriminated along two dimensions, predominantly relating to nutritional value ("healthy", "fatty", "fattening") and sensory/functional characteristics ("versatile", "bland", "boring", "a meal in itself"). Along with further analysis of the sensory descriptors, these results indicate the utility and efficiency of RGM for clarifying consumer views of broad food categories, while identifying the potential acceptability of particular starchy foods in fulfilling current dietary goals. PMID- 9218099 TI - A question of balance: nutrition, health and gastronomy. AB - Given the higher proportion of manufactured foods now available which meet current dietary recommendations, the food supply in developed countries like Australia could be said to be "healthier". Yet the "health" of the diet is often achieved at the expense of the "health" of the environment since ecological problems created a current food production and distribution methods remain unaddressed. Further, nutritional modifications which produce foods that are low in fat, sugar, salt and high in fibre do not necessarily address the concerns consumers have about the food supply. An emphasis solely on the physical health of populations, through improved diet, is out of keeping with current views on health which recognise the importance of overall well-being. Through the development of the concept of "sustaining gastronomy", consumers, food manufacturers and producers, and food regulators can better address the problems inherent in the food system, including those of an environmental nature. PMID- 9218100 TI - The insulin story: a 25-year perspective. PMID- 9218102 TI - Detection of Kell blood groups: molecular methods in the diagnostic laboratory. AB - The introduction of molecular biology techniques to the field of transfusion medicine has allowed more detailed analysis of the basis for the expression of several blood-group antigens. The application of the polymerase chain reaction for the determination of red-cell blood-group genotype is a highly sensitive technique that can be used to determine the likelihood of a fetus being affected by haemolytic disease of the fetuses or newborn. This alone makes polymerase chain reaction based techniques highly desirable for such applications. The determination of the genetic basis for the Kell/Cellano polymorphism has enabled the development of polymerase chain reaction-based techniques for genotyping. Several other red-cell blood-group antigen polymorphisms can also be analysed in this way. PMID- 9218101 TI - New drugs in essential thrombocythemia and polycythemia vera. AB - Among the chronic myeloproliferative disorders, polycythemia vera and essential thrombocythemia are unique because of their association with thrombohemorrhagic manifestations and their relatively indolent clinical course. Patients with essential thrombocythemia may not have a significant shortening of life expectancy and most may not require specific therapy. However, patients with polycythemia vera have a significant risk of transformation of polycythemia vera into acute leukemia or postpolycythemic myelofibrosis (or both). 'High-risk-for thrombosis' patients with either polycythemia vera or essential thrombocythemia require specific therapy with a platelet-lowering agent to prevent thrombotic complications. Currently, the standard agent used for this is hydroxyurea. However, its tetratogenic and leukemogenic potential has been of concern. As a result, new platelet-lowering agents are being evaluated in the treatment of polycythemia vera and essential thrombocythemia. Anagrelide and interferon alfa are two such agents and have been shown to be effective in reducing platelet counts in patients with chronic myeloproliferative disorders. The putative mechanism of action of these drugs, their specific activity in polycythemia vera and essential thrombocythemia, side-effect profile, and current indications are discussed herein. PMID- 9218103 TI - Use of haemopoietic growth factors: commentary on the ASCO/ECOG guidelines. American Society of Clinical Oncology/Eastern Co-operative Oncology Group. AB - In the past two to three years, a number of clinical guidelines have been issued that have attempted to recommend specific indications for the use of haemopoietic colony-stimulating factors. Some of which have recently been updated. These guidelines are, for the most part, a welcome initiative to guide clinicians as to the most appropriate use of these powerful and expensive drugs. Here, we have attempted to offer a perspective on these guidelines based upon both a review of the literature and our own clinical experience, concentrating primarily on their use following conventional and high-dose therapy and for the purpose of peripheral blood stem cell mobilization. For the latter, it is worth remembering that a product licence for this indication was only obtained in Europe in 1995 and that this only covers autologous mobilization and not the mobilization of normal donors. Both the American Society of Clinical Oncology and the Eastern Co operative Oncology Group guidelines rightly confine their comments to best practice. However, certainly in the United Kingdom and probably elsewhere, it is impossible to divorce this entirely from the economic realities involved in the widespread use of colony-stimulating factors, hence the increasing number of research papers in which the authors emphasize not only the clinical but also the financial implications of their findings. Here, we have taken note of both clinical and economic considerations in framing our comments. PMID- 9218105 TI - High-dose cytosine arabinoside in the treatment of acute myeloid leukaemia. AB - Cytosine arabinoside (Ara-C) has earned its recognition as the most important antileukaemic drug currently available for the treatment of acute myeloid leukaemia. Approximately 60-80% of patients less than 60 years of age can be expected to enter complete remission if treated with standard-dose Ara-C (100-200 mg/m2) in combination with an anthracycline. The efficacy of Ara-C appears clinically to be dose and schedule dependent. A 15-30 fold dose escalation in Ara C can elicit a therapeutic response in patients who have previously failed treatment with standard-dose Ara-C or other anti-leukaemic drugs. The use of high dose cytosine arabinoside (HDAC 3 gm/m2) appears rational based on cytosine pharmacology. Drug-scheduling is used to exploit drug synergy when HDAC is given in combination with asparaginase or fludarabine (+/- G-CSF) in a schedule dependent fashion. Toxicity following Ara-C is also dose- and schedule-dependent. Central nervous system toxicity--particularly cerebellar dysfunction--although rare, is particularly serious because it may preclude further use of the drug. Older patients are particularly susceptible. This article will describe the rationale for and the value of HDAC alone or in combination with other cytotoxics in the treatment of acute myeloid leukaemia. PMID- 9218104 TI - Expression of adhesion molecules in malignant plasma cells in multiple myeloma: comparison with normal plasma cells and functional significance. AB - Malignant plasma cells in multiple myeloma are predominantly confined to the bone marrow, where they stimulate cytokine production by stromal cells and bone cells leading to osteoclast activation and formation of the characteristic lytic lesions in the skeleton. Adhesion molecules are critically involved in the cellular interactions between myeloma cells and stromal elements and may represent novel therapeutic targets to reduce osteolytic bone disease in multiple myeloma. Here, we review the literature on the adhesion molecule repertoire expressed by malignant plasma cells and discuss the evidence that adhesive interactions between myeloma cells and stromal cells stimulate production of bone resorbing cytokines. PMID- 9218106 TI - Monosomy 7 and 7q--associated with myeloid malignancy. AB - An association between the complete or partial loss of chromosome 7 and preleukaemic myelodysplasia or acute myeloid leukaemia has been recognized from the early days of tumour cytogenetic analysis. Detection of such abnormalities usually heralds a poor prognosis. The loss of DNA on chromosome 7 has led to speculation that tumour-suppressor genes may play a significant role in this form of leukaemogenesis, although it may be part of a multistep process. A further association with leukaemia secondary to carcinogen exposure including previous chemotherapy or a number of congenital anaemias has increased the interest in discovering the gene or genes on chromosome 7. Banded chromosome analysis has suggested that there are two broad critical regions on the long arm of chromosome 7 at bands 7q22 and 7q34-q36 that may contain the relevant genes. Initial molecular analysis has confirmed these two regions to be of significance. The advent of fluorescence in-situ hybridization techniques has facilitated some definition of the 7q22 region, with identification of candidate genes for further functional analysis. It is becoming clear that there will be more than one gene on chromosome 7 involved in the leukaemic process and with the definition of these genes it may be possible to look for associations with different phenotypes and prognosis. As for the reason for chromosome 7 showing a particular predisposition to total or partial loss we may speculate that the DNA sequence and structure may confer a 'fragility' on the chromosome. A greater understanding of the DNA structure of the long arm may provide real insight into the mechanisms of leukaemia. We would like to speculate in the long term that this could lead to the ability to screen for leukaemia susceptibility and avoidance of 'inducers' in those at risk. PMID- 9218107 TI - Finite element analysis of the cranial base in subjects with Class III malocclusion. AB - The association between cranial base morphology and Class III malocclusion is poorly understood. This study analyses local shape- and size differences in cranial base configurations of Class I and Class III subjects, employing finite element (FEM) analysis. Seventy-three prepubertal European-American children with Class III malocclusion were compared to their counterparts with a normal, Class I molar occlusion. Lateral cephalographs were traced, checked and subdivided into age- and sex-matched groups. Thirteen points on the cranial base were identified and digitized, providing a geometrical cranial base representation. Average cranial geometries were scaled to an equivalent size and a FEM analysis, capable of depicting and quantifying local shape- and size-change, employed to highlight regionalized, morphological differences. While the anterior cranial base was more homogeneous for shape-change, significant, localized anisotropy in the posterior regions of the cranial base and around sella turcica was evident. For size change, areas of negative allometry were located posteriorly, but dilations in the mid- and anterior cranial base also were apparent. It is concluded that morphological alterations within the petro-occipital complex accompanied by changes in the sphenoidal and ethmoidal regions induce deviation from a normal cranial base configuration to one associated with deficient orthocephalization and an appearance of Class III malocclusion. PMID- 9218108 TI - A cautionary tale of simplified retention. AB - Attention is drawn to a possible serious unwanted sequela of a simplified bonded retainer. It is illustrated by three brief case histories. PMID- 9218109 TI - The orthodontic management of patients with profound learning disability. AB - The orthodontic management of patients with profound learning disability is described including a report of two cases. Close co-operation between the orthodontist and other providers of dental treatment is essential for the management of this group of patients, and a general anaesthetic may be needed to allow fixed appliances to be placed. The orthodontic treatment is co-ordinated with any necessary restorative or periodontal treatment. The moral and ethical questions raised by the treatment offered to this group of patients are discussed. PMID- 9218110 TI - An investigation into the placement of force delivery systems and the initial forces applied by clinicians during space closure. AB - This in vitro investigation was designed to establish not only how clinicians apply forces for space closure when using the straight wire appliance and sliding mechanics, but also to quantify the initial force levels produced. A single typodont, with residual extraction space in each quadrant, was set up to simulate space closure using sliding mechanics. On two occasions, at least 2 months apart, 18 clinicians were asked to apply three force delivery systems to the typodont, in the manner in which they would apply it in a clinical situation. The three types of force delivery system investigated were elastomeric chain, an elastomeric module on a steel ligature, and a nickel-titanium closed coil spring. A choice of spaced or unspaced elastomeric chain produced by a single manufacturer was provided. The amount of stretch which was placed on each type of system was measured and, using an Instron Universal Testing Machine, the initial force which would be generated by each force delivery system was established. Clinicians were assessed to examine their consistency in the amount of stretch which each placed on the force delivery systems, their initial force application and their ability to apply equivalent forces with the different types of force delivery system. The clinicians were found to be consistent in their method of application of the force delivery systems and, therefore, their force application, as individuals, but there was a wide range of forces applied as a group. However, most clinicians applied very different forces when using different force delivery systems. When using the module on a ligature the greatest force was applied, whilst the nickel titanium coil springs provided the least force. PMID- 9218112 TI - The design and analysis of reliability studies for the use of epidemiological and audit indices in orthodontics. PMID- 9218111 TI - Alternatives to ceramic brackets: the tensile bond strengths of two aesthetic brackets compared ex vivo with stainless steel foil-mesh bracket bases. AB - The mean tensile/peel bond strengths were evaluated for three types of aesthetic brackets (a ceramic-reinforced bracket and two generations of a ceramic/polycarbonate combination bracket). These were found to be significantly lower than the mean tensile/peel bond strength of a convention foil-mesh stainless steel bracket base. Failure of the ceramic-reinforced polycarbonate brackets occurred predominantly by fracture of the tie wings during testing. With the ceramic/polycarbonate combination brackets, the majority of the specimens failed due to separation of the ceramic and polycarbonate parts of the bracket. PMID- 9218113 TI - Severe Infra-occlusion and failed eruption of deciduous molars associated with eruptive and developmental disturbances in the permanent dentition: a report of 28 selected cases. AB - Retrospective analysis of 28 children suffering from severe infra-occlusion and/or primary failure of eruption of deciduous molars revealed an association with eruptive and developmental disturbances in the permanent dentition, including ectopically placed teeth and aplasia of teeth. Taurodont permanent molars were evident in 19 of the 28 selected cases which suggests a possible developmental relationship between these factors. Problems in relation to treatment of these cases are discussed. PMID- 9218114 TI - A combined orthodontic/restorative clinic. Rationale, evolution, and experience. AB - The rationale for running combined clinics between orthodontics and restorative dentistry is given, together with the history of the development and experience of such a clinic at the Dental School and Hospital in Cardiff. Brief details of the organization of the combined clinic and possible future developments are also given. PMID- 9218115 TI - A survey of the opinions of orthodontic specialist trainees in the U.K. AB - A questionnaire survey was carried out to ascertain a profile of orthodontic postgraduates in training in the United Kingdom during 1993. Information about the postgraduates, their programmes and their career plans was collected. Eighty nine questionnaires were distributed to those enrolled in 13 of the training programmes in the U.K. at that time from which the response rate was 64 per cent. The results can be compared with a similar survey carried out in the United States of America in 1992 (Keith and Proffit, 1994). PMID- 9218116 TI - The development of a speciality training programme. PMID- 9218117 TI - Bracket recycling--who does what? AB - At present there is debate concerning the practice of recycling orthodontic components. A survey of 300 members of the British Orthodontic Society showed that 47.5 per cent of respondents recycled metal brackets and that more specialist practitioners than consultants did so (p < 0.001). Only 7.2 per cent of the orthodontists who recycled brackets informed their patients that recycled brackets were used. PMID- 9218118 TI - Maternity rights and the law. AB - This article outlines the main points relating to Maternity Rights and the law. It emphasises the wisdom of being knowledgeable of these aspects, and ensuring they are understood by relevant members of staff. PMID- 9218119 TI - The 'hands free' method for the accurate placement and bonding of a multistrand wire splint. PMID- 9218120 TI - An evaluation of strategies available for the identification of GTP-binding proteins required in intracellular signalling pathways. AB - Strategies which can be used to elucidate the nature of a GTP-binding regulatory protein (G-protein) involved in an intracellular pathway of interest in the complex environment of the cell are described and evaluated. A desirable strategy is considered to be one in which the first stage indicates a requirement for one or more G-proteins, provides information on whether a monomeric, trimeric or other type of G-protein is involved, and gives some idea of the G-protein sub class. In the second stage the specific G-protein involved is identified. Approaches available for investigations in the first stage include the use of analogues of GTP and GDP, AlF4-, inhibitors of G-protein isoprenylation, bacterial toxins which covalently modify G-proteins, and the introduction of a purified GDP dissociation inhibitor, GDP exchange and/or GTP-ase activating protein. Identification of the specific G-protein in the second stage can be achieved using anti G-protein antibodies, G-protein-or receptor-derived peptides, antisense G-protein RNA and over-expressed, constitutively-active or dominant negative G-protein mutants. The correct interpretation of results obtained with GTP and GDP analogues and AlF4- in the first stage is complex and often difficult, and requires a thorough understanding of the functions and mechanisms of activation of G-proteins. Nevertheless, it is important to reach the correct conclusion at this stage since considerable time and expense are usually required for investigations in the second stage. PMID- 9218121 TI - Glutamate receptors and gene induction: signalling from receptor to nucleus. AB - Activation of glutamate receptors has been linked to a diversity of lasting physiologic and pathologic changes in the mammalian nervous system. The cellular and molecular mechanisms underlying permanent modifications of nervous system structure and function following brief episodes of neuronal activity are unknown. Immediate early genes (IEGs) have been implicated in the conversion of short-term stimuli to long-term changes in cellular phenotype by regulation of gene expression. Many of the long-term consequences of glutamate receptor activation correlate with increases in specific IEGs; the intracellular signalling pathways coupling activation of receptors at the cell surface with induction of IEGs in the nucleus are incompletely understood. Analysis of mechanisms of how extracellular factors control gene expression implicate activation of second messenger systems and protein kinases. Activation of glutamate receptors results in an initial increase in intracellular calcium; the route of calcium influx may differ depending on the specific receptor subtype activated. Intracellular calcium is often the first messenger in response to an extracellular stimulus and can be the trigger for activating numerous other signalling pathways. Results obtained over the past several years support a hypothesis where selective activation of distinct intracellular signalling pathways and IEG responses, following activation of different glutamate receptor subtypes, involve spatial restriction of key enzymes to sites of local calcium increases. The specificity in long-term neuronal responses following brief synaptic activation may depend on the specific intracellular signalling mechanisms triggered and the unique array of IEGs transcribed. PMID- 9218122 TI - Proposal for pharmacologically distinct conformers of PDE4 cyclic AMP phosphodiesterases. AB - cAMP-specific phosphodiesterase inhibitors display a range of activities in vitro and in vivo which suggest they may be useful in the treatment of inflammatory diseases. However, these compounds elicit a number of side-effects which may limit their therapeutic potential. Certain side-effects of PDE4 inhibitors such as emesis and gastric acid secretion are associated with their actions at a high affinity rolipram binding site (HARBS). In contrast, a number of anti inflammatory actions of PDE4 inhibitors are better correlated with inhibition of PDE4 catalytic activity than with displacement of [3H] rolipram from HARBS. This suggests that native PDE4s in different cell-types can be discriminated pharmacologically. Although known to be associated with PDE4, the nature of HARBS is uncertain. The majority of evidence suggests it represents particular conformational states of PDE subtypes with which rolipram interacts with high potency (KD approximately 2 nM) (High-affinity PDE4, HPDE4). Rolipram is generally moderately or weakly active (IC50-200 nM-2000 nM) in inhibiting catalytic activity of the majority of crude, partially-purified or recombinant PDE4-preparations (Low-affinity PDE4, LPDE4). Solubilization or V/GSH treatment of particulate eosinophil PDE4, cAMP-dependent kinase activation of RNPDE4D3 and membrane association of HSPDE4A4 increase the potencies of some (e.g., rolipram) but not other (e.g., trequinsin) inhibitors. In eosinophils, the changes in enzyme properties brought about by solubilization result in a close correlation between the potency order of compounds in inhibiting cAMP hydrolysis and displacing [3H] rolipram from HARBS. The identification of distinct pharmacological PDE4 forms may have therapeutic consequences since it may be possible to synthesize potent inhibitors of LPDE4 with low affinity for HARBS which should, theoretically, be less emetic. Most inhibitors synthesized to date (rolipram, denbufylline nitraquazone, etc.) display high-affinity for HARBS but are much weaker in inhibiting cAMP hydrolysis. Other compounds (RP 73401, trequinsin, CDP 840) display slightly higher potency against LPDE4 or do not discriminate between the two putative PDE4 forms. Recently, inhibitors have been synthesized which are considerably more active against LPDE4 than HPDE4. Such compounds with appropriate pharmacokinetic properties may retain anti inflammatory activity but have a reduced capacity to cause nausea and emesis and, consequently, have a wider therapeutic window than compounds currently undergoing clincial evaluation. PMID- 9218123 TI - G proteins and opioid receptor-mediated signalling. AB - Most opioid receptor-mediated functions appear to be mediated through G protein interactions, therefore an understanding of opioid signalling requires knowledge of those interactions. This review chronicles the studies examining these interactions for all the opioid receptor subtypes, both in vivo and in vitro. PMID- 9218124 TI - Oncoprotein signalling and mitosis. AB - Studies of the roles of oncoproteins in cell cycle progression have concentrated on G1 because transformation is frequently associated with loss of G1 checkpoint control. However, it has become evident that G2 and mitotic checkpoints are often compromised in transformed cells and that many tumour suppressor proteins and oncoprotein kinases regulate and/or are activated in G2 and M. Disruption of p53 and ATM tumour suppressor protein functions can eliminate G2 and M checkpoints. The Src family kinases are activated in mitosis and collectively play an indispensable role in progression through G2/M. In addition, evidence suggests that Mos and elements of the Ras/Raf/MAPK cascade are also active in mitosis and appear likely to regulate G2 and/or M. Potential targets of these kinases include likely regulators of gene expression and microtubule dynamics such as Sam68 and Oncoprotein 18/stathmin. The ability of some oncoproteins to perturb orderly progression through both G1 and/or S and G2 and/or M is probably important for transformation. PMID- 9218125 TI - Phosphatidic acid-mediated regulation of neutrophil plasma membrane CD45 phosphotyrosine phosphatase. AB - CD45-phosphotyrosine phosphatase (PTPase) constitutes the major portion of thr PTPase activity within plasma membranes of neutrophilic leukocytes, where it regulates signals leading to functional activation. We have previously demonstrated that the catalytic component of neutrophil plasma membrane CD45 PTPASE is regulated by a cytosolic inactivator which itself is attenuated upon cellular stimulation, allowing enzyme translocated from granule stores to express full activity. The present study investigated mechanisms of cytosolic inactivator attenuation. Preincubation of plasma membranes of stimulated neutrophils with cytosol from resting cells resulted in a rapid loss of membrane-associated PTPase activity. Phosphatidic acid had no direct effect on plasma membrane PTPase activity but blunted in a dose dependent manner the effects of the PTPase inactivator. Inactivator attenuation was not observed with equivalent concentrations of either diacylglycerol or lysophosphatidic acid. Optimal attenuation of inactivator activity was obtained with long chain, soluble ligands, such as dicapryl phosphatidic acid. Inhibitors of neutrophil plasma membrane ecto-phosphatidic acid phosphohydrolase did not block inactivator attenuation, suggesting that phosphatidic acid and not one of its metabolites was the entity responsible. In conclusion, neutrophil plasma membrane PTPase is dynamically regulated by a cytosolic inactivator, the inhibition of which may potentiate the effects of PTPase translocated during cellular stimulation. Phosphatidic acid generated as a consequence of cellular stimulation may mediate this inhibition and thereby regulate the effects of tyrosine kinases activated during the initial phases of cell stimulation. PMID- 9218126 TI - Stimulation of insulin release by D-glucose in depolarized pancreatic islets. AB - This study reevaluates the relevance of plasma membrane depolarization in the pancreatic islet B-cell to the stimulation of insulin release by D-glucose. In rat pancreatic islets exposed to increasing concentrations of D-glucose, a sigmoidal relationship is observed between the output of insulin and concentration of the hexose. The release of insulin recorded in the absence of D glucose or at a non-stimulatory concentration (2.5 mM) of the hexose is significantly enhanced when the islets are incubated in the presence of the meglitinide analogue A-4166 (20 microM) or at a high concentration (30 microM) of extracellular K+. Nevertheless, even in the presence of A-4166 or at the high K+ concentration, increasing concentrations of D-glucose, up to 20 microM, still enhance, in a sigmoidal manner, the secretion of insulin, with a 15-fold or greater increment in insulin output relative to the value found in the absence of the hexose. These findings indicate that D-glucose provides a concentration related signal for insulin release, that cannot be attributed solely to either the closing of ATP-sensitive K+ channels, the depolarization of the plasma membrane, or the role of D-glucose as a nutrient to cover the energy expenditure in the islet cells. PMID- 9218127 TI - Signal transduction in gastrointestinal smooth muscle. AB - Signal transduction in gastric and intestinal smooth muscle is mediated by receptors coupled via distinct G proteins to various effector enzymes, including PI-specific PLC-beta 1 and PLC-beta 3, and phosphatidylcholine (PC)-specific PLC, PLD and PLA2. Activation of these enzymes is different in circular and longitudinal muscle cells, generating Ca(2+)-mobilizing (IP3, AA, cADPR) and other (DAG) messengers responsible for the initial and sustained phases of contraction, respectively. IP3-dependent Ca2+ release occurs only in circular muscle. Ca2+ mobilization in longitudinal muscle involves a cascade initiated by agonist-induced transient activation of PLA2 and formation of AA, AA-dependent depolarization of the plasma membrane and opening of voltage-sensitive Ca2+ channels. The influx of Ca2+ induces Ca2+ release by activating sarcoplasmic ryanodine receptor/Ca2+ channel and stimulates cADPR formation which enhances Ca(2+)-induced Ca2+ release. The initial [Ca2+]i transient in both muscle cell types results in Ca2+/calmodulin-dependent activation of MLC kinase, phosphorylation of MLC20 and interaction of actin and myosin. The sustained phase is mediated by a Ca(2+)-independent isoform of PKC, PKC-epsilon DAG for this process is generated by PLC- and PLD-mediated hydrolysis of PC. Relaxation is mediated by cAMP-and/or cGMP-dependent protein kinase which inhibit the initial [Ca2+]i transient and reduce the sensitivity of MLC kinase to [Ca2+]i. Relaxation induced by the main neurotransmitters, VIP and PACAP, involves two cascades, one of which reflects activation of adenylyl cyclase. A distinct cascade involves G protein-dependent stimulation of Ca2+ influx leading to Ca2+/calmodulin-dependent activation of a constitutive eNOS in muscle cells; the generation of NO activates soluble guanylyl cyclase. The resultant activation of PKA and PKG is jointly responsible for muscle relaxation. PMID- 9218128 TI - Hormonal responsiveness of hepatocytes after hypothermic preservation in University of Wisconsin solution. AB - The hormonal responsiveness of freshly isolated rat hepatocytes was compared to that of a) cold-preserved isolated hepatocytes and b) hepatocytes isolated from cold-preserved whole liver. Cold-preserved hepatocytes and cells isolated from cold-preserved whole liver increased phosphorylase alpha activity in response to norepinephrine (plus propranolol), vasopressin, angiotensin II and glucagon. However, the maximal response to these agents was smaller than that of freshly isolated hepatocytes. Basal phosphorylase alpha activity was increased in cold preserved hepatocytes. Similarly, cold preservation decreased the accumulation of cyclic AMP induced by glucagon and the effects of norepinephrine (plus propranolol), vasopressin and angiotensin II on the production of inositol phosphates. Basal levels of cyclic AMP were similar in the three conditions studied but basal production of [3H]IP2 plus [3H]IP3 was increased in cold preserved hepatocytes. There was a very small effect of beta-adrenergic activation on phosphorylase activity and a small accumulation of cyclic AMP in response to isoproterenol in the conditions studied. PMID- 9218129 TI - Cationic events stimulated by D-glucose in depolarized islet cells. AB - Pancreatic islets prelabelled with either 86Rb or 45Ca were exposed to a rise in D-glucose concentration from 2.8 to 16.7 mM whilst perifused in the presence of 2 microM glibenclamide, 30 mM extracellular K+ and both 30 mM K+ and 250 microM diazoxide. In all three situations, the rise in glucose concentration provoked a dramatic increase in insulin output, despite unchanged or even increased efflux of 86Rb from the prelabelled islets. Also in all three situations, glucose sharply decreased effluent radioactivity from islets prelabelled with 45Ca but perifused in the absence of extracellular Ca2+, while augmenting 45Ca efflux, to a variable extent, from islets perifused at normal extracellular Ca2+ concentration (1.0 mM). It is proposed, therefore, that the insulinotropic action of D-glucose in depolarized islets, and presumably also under normal conditions, may involve the gating of voltage-insensitive Ca2+ channels. PMID- 9218130 TI - Modifications of p53 protein and accumulation of p21 and gadd45 mRNA in TGF-beta 1 growth inhibited cells. AB - Transforming growth factory beta (TGF-beta) is a potent growth inhibitor of epithelial cells. One of the strategies used to elucidate the anti-proliferative mode of action of TGF-beta is to find out whether the receptor-generated signals interact with components of the basic machinery of the cell cycle. In this study we examined whether p53 and two other cycle inhibitory genes that can be transactivated by p53 are affected by TGF-beta 1 in epithelial cells. We show that TGF-beta 1 signalling controls the intracellular localization as well as the phosphorylation pattern and the stability of p53 protein. TGF-beta signalling also elevates the expression of p21/waf-1 and gadd45. The observed modifications in the protein suggest that p53 is involved in mediation of TGF-beta 1 growth inhibition. However, in TGF-beta 1 growth inhibited cells, wild type p53 is not required for the accumulation of the two p53 downstream targets p21/waf-1 and gadd45. PMID- 9218131 TI - Annexin II binds to the membrane of A549 cells in a calcium-dependent and calcium independent manner. AB - We investigated the nature of annexin II binding to the biological membranes using a lung epithelium-derived cell line A549. The cytosolic and membrane fractions of A549 cells were separated in the presence of 5 mM EGTA. Both fractions contain annexin II monomer and tetramer as evaluated by western blots using specific monoclonal antibodies against p36 and p11 subunits of annexin II. A substantial amount of annexin II was associated with the membrane fraction even after extensive washing with EGTA buffer, indicating the presence of two pools of annexin II. The EGTA-resistant membrane-bound annexin II could be partially extracted by 1% Triton X-100 or 60 mM n-octyl-beta-D-glucopyranoside, and completely by 30 mM CHAPS or 0.1% deoxycholate. This fraction of annexin II was also extracted by 0.1 M Na2CO3, pH 11 and partitioned into the aqueous phase after being treated with Triton X-114, demonstrating that the EGTA-resistant annexin II is a peripheral membrane protein. When the cells were lysed in varying concentrations of Ca2+, annexin II translocated from cytosolic fraction to membrane fraction at 4-25 microM Ca2+. To identify proteins closely associated with annexin II the membrane fraction was treated with the bifunctional chemical cross-linker disulfosuccinimidyl tartarate, followed by western blot analysis using anti-p36 or anti-p11 antibodies. We find that both p36 and p11 were cross linked to a 51 kDa protein. In addition, p11 also binds to several proteins with molecular mass of 91, 65, 40 and 36 kDa. Our results suggest that annexin II may bind to the A549 cell membranes via specific membrane-associated proteins. PMID- 9218132 TI - Role of phosphatidylinositol 3-kinase in degranulation induced by IgE-dependent and -independent mechanisms in rat basophilic RBL-2H3 (ml) cells. AB - We have examined the role of phosphatidylinositol 3-kinase (P13-kinase) in the degranulation induced by the antigen, an IgE-dependent stimulant, and by carbachol and thapsigargin, IgE-independent stimulants, in the muscarine ml receptor-transfected mast cell line RBL-2h3 (ml) cells. These stimulants commonly increased P13-kinase activity in the anti-phosphotyrosine immunoprecipitate. The P13-kinase inhibitors wortmannin and LY294002 inhibited induced by these stimulants. The membrane ruffling induced by the antigen or carbachol was also inhibited by wortmannin. In contrast, thapsigargin induced by membrane ruffling but induced microspikes, which was not affected by wortmannin. In the permeabilized RBL-2H3 (ml) cells, wortmannin the GTP gamma S-induced membrane ruffling without inhibiting the GTP gamma S-induced degranulation. These findings suggest that P13-kinase is involved not only in IgE-dependent degranulation but also in IgE-independent degranulation, and that the GTP gamma S-sensitive protein at the downstream of P13-kinase is responsible for the degranulation but not for the membrane ruffling. PMID- 9218133 TI - Platelet-derived growth factor activates a mammalian Ste20 coupled mitogen activated protein kinase in airway smooth muscle. AB - We have investigated the mechanisms regulating p38MAPK in airway smooth muscle cells. Incubation of cells with platelet-derived growth factor (PDGF) stimulated MAPKA kinase-2 activity, a kinase immediately down-stream of P38MAPK. Preincubation of the cells with forskolin (10 microM), which stimulated a 20-fold increase in intracellular cAMP formation, inhibited this response. An antibody raised against subdomain VI of yeast Ste20 kinase co-immunoprecipitated p38MAPK from cell lysates. The immunoprecipitated kinase(s) was shown to catalyse the phosphorylation of myelin basic protein (MBP) and to activate purified MAPKAP kinase-2. Incubation of cells with PDGF did not increase the amount of p38MAPK isolated in the anti-Ste20 immunoprecipitate. However, the kinase phosphorylated MBP and stimulated purified MAPKAP kinase-2 activity more effectively than kinase from control cells. The preincubation of cells with forskolin (10 microM) reduced the amount of P38MAPK in the immunoprecipitate and this correlated with a decrease in kinase activity. We conclude the PDGF induces the activation of p38MAPK, whereas forskolin elicits its dissociation from the complex with Ste20. Therefore, Ste20/p38MAPK may form part of a signal transduction pathway linked to activation of MAPKAP kinase-2 in ASM cells. PMID- 9218134 TI - Negative regulation of MAP kinase by diacylglycerol-dependent mechanisms via G protein-coupled receptors in rat basophilic RBL-2H3 (ml) cells. AB - Carbachol and 5'-(N-ethylcarboxamido)-adenosine (NECA), stimulants of G protein coupled receptors, induce MAP kinase activation in the muscarinic ml receptor transfected mast cell line, RBL-2H3 (ml) cells. The phospholipase C inhibitor neomycin and the phosphatidate phosphohydrolase inhibitor propranolol augmented MAP kinase activation induced by carbachol and NECA without affecting the antigen induced MAP kinase activation. Furthermore, the duration of MAP kinase activation induced by carbachol or NECA was also prolonged by neomycin and propranolol. The specific protein kinase C inhibitor Ro 31-8425 enhanced the carbachol- or NECA induced MAP kinase activation. These findings suggest that the MAP kinase activation mediated by the G protein-coupled receptors is negatively regulated by diacylglycerol and activated protein kinase C(s). PMID- 9218135 TI - Cyclic AMP inhibitors inhibits PDGF-stimulated mitogen-activated protein kinase activity in rat aortic smooth muscle cells via inactivation of c-Raf-1 kinase and induction of MAP kinase phosphatase-1. AB - In rat aortic smooth muscle cells (RASMC), pretreatment with forskolin inhibited the activation of p42/44 isoforms of mitogen-activated protein kinase (MAP) kinase stimulated in response to low concentrations of PDGF (10 ng/ml). This correlated with a strong inhibition of PDGF-stimulated MEK and C-Raf-1 kinase activity. However, the effect of forskolin could be surmounted by increasing the concentration of PDGF. Under such conditions forskolin was only effective against prolonged MAP kinase activation. The ability of forskolin to inhibit the late phase of MAP kinase activity was reversed by pretreatment of the cells with cycloheximide, suggesting the involvement of a protein synthesis step. This was not due to effects upstream of MAP kinase since PDGF-stimulated MEK activation was decreased by cycloheximide, an effect potentiated by forskolin. Forskolin stimulated the induction of the dual specific phosphatase MAP kinase phosphatase 1 (MKP-1), although this effect was small relative to levels induced by PDGF and angiotensin II. However, PDGF stimulated induction of MKP-1 was abolished by the protein kinase A inhibitor H89 and this correlated with the reversal of forskolin mediated inhibition of PDGF-stimulated MAP kinase activity. These studies implicate a role for intracellular cyclic AMP in at least two aspects of MAP kinase signaling, including both the inhibition of Raf-1 activation and the induction of MKP-1. PMID- 9218136 TI - Odor hedonics: connection with emotional response estimated by autonomic parameters. AB - The aim of this paper is to analyse the relationship between self-report hedonic evaluations and the physiological expression of emotion in response to odorants. We try to solve the following questions: (1) Is it possible to find any experimental evidence that the sense of smell is linked with emotion? (2) What kind of odorants can be distinguished by autonomic analysis? (3) Is there a link between hedonics and autonomic information? The effects of odorants on the emotional process were estimated, in terms of autonomic nervous system (ANS) activity. Fifteen subjects inhaled five odorants as olfactory stimuli: lavender (LAV), ethyl acetoacetate (EAA), camphor (CAM), acetic acid (AA) and butyric acid (BA). After inhaling the odorant, subjects were requested to fill out an 11-point hedonic scale to rate its pleasantness versus unpleasantness. ANS parameters were as follows: two electrodermal responses, skin potential (SP) and resistance (SR); two thermovascular parameters, skin blood flow (SBF) and skin temperature (ST); and two cardiorespiratory parameters; instantaneous respiratory frequency (IRF) and instantaneous heart rate (IHR). Simultaneous recording of six parameters showed that specific autonomic patterns were associated with each odorant. An analysis of variance made it possible to differentiate among the five odorants. Two-by-two odorant comparisons for autonomic responses using Tukey's HSD multiple comparison test only permitted differentiation between pleasant odorants (LAV and EAA) and unpleasant (AA and BA) ones, but camphor was differentiated from both pleasant and unpleasant odorants. Each odorant elicited responses in the different parameters, yet subjects responded through their preferential channels; an average of two channels was used by each subject. These results when compared with those obtained with other senses (visual and auditory), did not evidence the postulated preferential link between olfaction and emotion. A strong link between hedonics and ANS response could be demonstrated when considering each subject and mainly through his/her preferential channel(s); conversely a weak correlation (SR duration excepted) was obtained between inter-subjects' hedonic evaluation. It seems that for a given population the autonomic response reflect the odor valence only through some parameters related to the main preferential channel(s) and thus the global autonomic pattern has to be considered. PMID- 9218137 TI - Capsaicin modifies responses of rat chorda tympani nerve fibers to NaCl. AB - Single-fiber preparations of the rat chorda tympani (CT) nerve were used to study the mechanism of action of capsaicin on salt-taste transduction. Capsaicin selectively suppressed the responses to NaCl of the CT nerve fibers (N-fibers) that are sodium-specific (insensitive or poorly sensitive to potassium). Among the more broadly responsive, cation-sensitive fibers (E-fibers) there are two subtypes, both of which responded to capsaicin but in different ways ('enhanced' type and 'suppressed' type). In both N- and E-fibers, 5% ethanol (the vehicle for capsaicin) slightly reduced the response to 100 mM NaCl. The suppressive effect of capsaicin on the response of the N-type fibers to 100 mM NaCl was significantly stronger than the effect of 5% ethanol. The suppression lasted for at least 20 s after the simultaneous application of 100 p.p.m. capsaicin-100 nM NaCl. These results indicate that 100 p.p.m. capsaicin can modify the response of CT fibers to NaCl. The observed effect of capsaicin on gustatory fibers could be the net result of opposite suppressive and enhancing processes in the taste buds cells and excited intra- or extragemmal trigeminal nerve endings. PMID- 9218138 TI - Perceived irritation during ingestion of capsaicin or piperine: comparison of trigeminal and non-trigeminal areas. AB - The aim of this study was to investigate the perception of chemosensory irritation in the oropharyngeal region during the ingestion of irritants. In two experiments subjects sipped and swallowed small samples of an ascending concentration series of capsaicin or piperine and rated the intensity of sensations or irritation perceived at four locations: the anterior tongue, the posterior tongue, the roof of the mouth and the throat. Both experiments revealed that the responsiveness to irritation from capsaicin was significantly higher in the throat than at either the front or back of the tongue. There was no difference between irritation ratings for the throat and the roof of the mouth. Compared with capsaicin, the responsiveness to piperine was more uniform along the rostro-caudal axis; for example, irritation ratings for the throat were similar to those for the anterior tongue. These results support previous findings which indicated that the oral mucosae were not uniformly sensitive to chemical irritants, and suggest further that the throat, which is innervated by both the glossopharyngeal and vagus nerves, plays an important role in the perception of chemesthetic stimuli during ingestion. PMID- 9218139 TI - Amiloride effects on taste quality: comparison of single and multiple response category procedures. AB - Although there is compelling evidence that amiloride reduces the intensity of Na+ and Li+ salts in humans, its effects on saltiness are conflicting. Many salts elicit not only a salty taste but also one or more side tastes (sweetness, sourness or bitterness). Some studies have shown a suppression of saltiness by amiloride; others show no effect on saltiness but a significant reduction in sourness. In the experiments demonstrating a reduction of saltiness, subjects estimated only saltiness; in those showing an amiloride effect on sourness and not saltiness, subjects estimated all qualities on each trial. The present study examines the role of the psychophysical method in these conflicting results. We have investigated the effects of amiloride on taste quality by modifying only the instructions to the subjects, keeping all other variables constant. One group of subjects (intensity-only) gave magnitude estimates of the overall intensity of a LiCl concentration series. A second group (salty-only) was instructed to estimate the saltiness of the stimuli, and a third group (sour-only) estimated their sourness. Finally, a fourth group (profile) rated all of the taste qualities on each stimulus presentation, using a modified taste profile method. The ratings of all groups were made comparable by the use of 0.1 mM quinine-HCl as a modulus. When subjects used only one response category, amiloride reduced their estimates (of intensity, saltiness or sourness), but if subjects attended to all four qualities, amiloride specifically reduced the sourness of LiCl and had no significant effect on its saltiness. Comparison of the saltiness estimates of the salty-only group to the sum of the salty and sour estimates of the profile group demonstrated that subjects combined these sensations when presented with only one response alternative. To reveal the effect of amiloride on a specific quality of a salt, the psychophysical method must allow subjects to attend to all qualities on each trial. These data and previous results suggest that apical Na+ channels on the taste receptor cell membrane mediate the sourness but not the saltiness of Na+ and Li+ salts. PMID- 9218140 TI - Rapid classical conditioning of odor response in a physiological model for olfactory research, the tiger salamander. AB - In recent years there have been a number of important advances in the understanding of cellular mechanisms related to olfactory function. Hypotheses regarding the complex relationships among odorant structure, physiological activity and behavioral outcome generated by these findings, however, remain largely untested due to a paucity of psychophysical data on stimulus discrimination in the same experimental species. Comparisons between behavioral and physiological responses are essential for elucidating the critical aspects of stimulus coding in sensory systems. We have developed a method for generating psychophysical data in one of the primary model species used in olfactory research, the tiger salamander, Ambystoma tigrinum. These psychophysical experiments are carried out under the same conditions as physiological experiments in our laboratory. Using classical conditioning, individual salamanders are trained over a period of 2-3 h to show skin potential responses to odor and not air. Failure to train using backward pairing demonstrates that the response is not due to sensitization or pseudoconditioning. The conditioned response is mediated by the olfactory pathway, as it is blocked by olfactory nerve section. We show that salamanders detect three odorants that are commonly used stimuli in physiological experiments (butyl alcohol, butyl acetate and amyl acetate), but cannot detect a fourth common experimental odorant, camphor. This method should be a powerful tool for studying olfactory information processing by providing data on discriminability of stimuli used in salamander physiological studies. PMID- 9218141 TI - Broad tuning of rat taste cells for four basic taste stimuli. AB - The breadth of responsiveness of rat taste cells to the four basic taste stimuli was studied using the entropy measure (H) proposed by Smith and Travers. H values range from 0.0 for narrow tuning to 1.0 for broad tuning. Based on the responses of depolarizing receptor potentials of 26 rat taste cells to the four basic taste stimuli, taste cells were classified into nine NaCl-best, four Q-HCl (quinine HCl)-best, 10 HCl-best and three sucrose-best cells. NaCl-best cells were narrowly tuned to the four basic taste stimuli, but the other three stimuli-best cells were broadly tuned to the stimuli. In all, 85% of the taste cells responded to more than one of four basic taste stimuli. The mean H values for NaCl-best, Q HCl-best, HCl-best and sucrose-best cells were 0.285, 0.832, 0.781 and 0.796 respectively. The mean H value for all 26 taste cells was 0.621. This was larger than H in rat gustatory fibers. Transformation of large H values in taste cells into small H values in taste fibers may be due to a non-random interaction between taste cells and taste fibers during the synaptic formation. Broad tuning properties of rat taste cells suggest that the across-taste cell response pattern may play an important role in taste quality coding mechanisms. PMID- 9218142 TI - Ultrastructural aspects of olfactory signaling. AB - The olfactory area of the nasal cavity is lined with olfactory receptor cell cilia that come in contact with incoming odor molecules. Ultrastructural immunocytochemical studies in rodents have shown that these cilia contain all the proteins necessary to transduce the odorous message into an electrical signal that can be transmitted to the brain. These signaling proteins include putative odor receptors, GTP binding proteins, type III adenylyl cyclase and cyclic nucleotide-gated channels. The rest of the cells, including dendrites and dendritic knobs, showed no discernible labeling with antibodies to these signaling proteins. Furthermore, freeze-fracture and freeze-etch studies have shown that the membrane morphology of olfactory cilia differs substantially from that of non-sensory cilia. Olfactory cilia have many more membrane particles. Transmembrane signaling proteins, such as odor receptors, adenylyl cyclase and cyclic nucleotide-gated channels, conceivably appear as membrane particles. Thus, the long-standing supposition that olfactory cilia are peculiarly adapted to deal with the reception and initial transduction of odorous messages has now been verified in terms of both ultrastructural morphology and cytochemistry. Emerging studies on vomeronasal receptor cell microvilli indicate that the same is true for this organ, even though the actual signaling components differ from those of the main olfactory system. PMID- 9218143 TI - Glucocorticoid (type II) receptors in the olfactory mucosa of the guinea-pig: RU 28362. AB - The glucocorticoid RU 28362 was employed to identify glucocorticoid receptors in the olfactory mucosa of the guinea-pig. Results demonstrate significant binding of RU 28362 and suggest that the olfactory mucosa is a target site for glucocorticoid action. PMID- 9218144 TI - Olfactory receptor database (ORDB): a resource for sharing and analyzing published and unpublished data. AB - An olfactory receptor database (ORDB) is being developed to facilitate analysis of this large gene family. ORDB currently contains over 400 olfactory receptor sequences and related information, and is available via the World Wide Web. We plan to incorporate functional data, structural models, spatial localization and other categories of information, toward an integrated model of olfactory receptor function. PMID- 9218145 TI - Predictive value of MCV for hazardous drinking in the community. AB - A recent study suggested that general practitioners (GPs) do not see the necessity of investigating MCVs which unexpectedly and only slightly exceed the reference limit, despite the association between MCV and alcohol abuse. Because a literature search could not find a study of the predictive value of the MCV for hazardous drinking in the community, such a study was undertaken among a random sample of 338 adults living in a regional Australian city. Twenty-nine of the adults admitted drinking hazardously. The MCV with the optimum sensitivity and specificity for identifying the hazardous drinkers was determined. An MCV of > 94 fl identified as many as 35% of the hazardous drinkers whilst misclassifying only 6% of the non-hazardous drinkers. The predictive value was even greater among males, 67%. We conclude that inquiring into MCVs > 94 fl will lead to GPs identifying a significant proportion of adults in the community admitting to hazardous drinking. PMID- 9218146 TI - Automated reticulocyte counting: evaluation of the Coulter STKS Haematology Analyser reticulocyte counting function. AB - This study evaluated reticulocyte counting with the automated reticulocyte function of the Coulter STKS Haematology Analyser. This is an upgrade option for Coulter STKS and MAXIM haematology analysers. Reticulocyte counts obtained with the automated reticulocyte counting function were compared with those obtained by visual counting. Reticulocyte counting with both methods gave excellent comparability with a correlation coefficient of 0.98. Results were consistent with the well documented imprecision of the manual method with a coefficient of variation (CV) of 16-22%. In contrast, the automated reticulocyte counting function was more precise with a CV of 12.3%. In both cases, counts were stable after storage for 24 h at room temperature and 4 degrees C. Our results suggest that the use of this upgrade will be beneficial for many laboratories. PMID- 9218147 TI - Serum transferrin receptor levels in patients undergoing evaluation of iron stores: correlation with other parameters and observed versus predicted results. AB - Serum transferrin receptor (sTfR) concentrations were measured in specimens from 77 patients undergoing serum ferritin determination, and the results correlated with serum ferritin, serum iron, serum total iron-binding capacity (TIBC) saturation, erythrocyte mean corpuscular volume (MCV), and mean corpuscular haemoglobin (MCH). All parameters exhibited the expected inverse correlation with sTfR; this correlation was statistically significant for all parameters except serum iron concentration. The frequency with which iron deficiency (defined as absence of stainable marrow iron) is observed in patients with particular ferritin values in this centre was determined and used to estimate the expected number of iron deficient patients in the present study. In no setting were significantly fewer sTfR levels > 3.05 micrograms/ml observed than expected. However, significantly greater than expected numbers of elevated sTfR values were observed in patients with serum ferritin > 220 micrograms/l (P = 0.002). The results suggest that the sTfR level is probably not useful as a single test for identification of iron deficiency in unselected patients. PMID- 9218148 TI - The spun micro-haematocrit and mean red cell volume are affected by changes in the oxygenation state of red blood cells. AB - In this study we examined the effects of in vitro oxygenation and deoxygenation on the spun micro-haematocrit (packed cell volume or PCV) and the electronically measured mean cell volume (MCV) and haematocrit (Hct) of red blood cells. Because PCV and/or MCV and Hct are being used for calibration of haematology analysers, it is important that their potential variability caused by outside factors is known. We found that these parameters did vary with the oxygenation state of the erythrocyte and conclude that the effect is most likely caused by a combination of water shifts due to intracellular carbamate and bicarbonate formation, and conformational changes in haemoglobin. The results of the study have implications for whole blood calibration and for short term imprecision assessments of automated haematology analysers. To ensure consistent results, we recommend that blood specimens be fully oxygenated before reference work or reproducibility studies of MCV, Hct and/or PCV are attempted. PMID- 9218149 TI - Successful treatment of autoimmune neutropenia with recombinant human granulocyte colony stimulating factor (R-metHuG-CSF). AB - Autoimmune neutropenia (AIN) is characterized by antibody mediated peripheral destruction of neutrophils. Since there is no effective treatment, antibiotics have to be used frequently for recurrent infections. Five selected patients with serologically proven AIN were treated with r-metHuG-CSF at 5-8 micrograms/kg body weight (300-480 micrograms) daily; the dose and frequency of r-metHuG-CSF was reduced after neutrophil counts above 1.0 x 10(9)/l were obtained. R-metHuG-CSF is effective in AIN and causes a sustained rise in ANC which can be maintained on a low dose administered twice or thrice weekly. PMID- 9218150 TI - A simple monoclonal antibody based ELISA for free protein S. Comparison with PEG precipitation. AB - A simple monoclonal antibody based ELISA for free Protein S, compatible with out existing ELISA for total Protein S has been developed, and its performance compared with the conventional PEG precipitation method of free Protein S assay. The normal range (mean +/- 2 SD) was 0.19-0.54 iu/ml free Protein S. The mean intra assay variation was 5.24% and the mean inter assay variation was 5.50%. A total of 102 routine diagnostic samples from patients referred for prothrombotic investigation (six assays for each method) were assayed by PEG precipitation (mean 0.32 iu/ml, SD 10.60), and the monoclonal ELISA (mean 0.34, SD 0.9). Paired t-test analysis of the two data sets indicated no significant difference between them (P < 0.001). In this sample population, there was no significant difference in free Protein S values when assayed by monoclonal based ELISA or by PEG precipitation. The monoclonal assay has proved to be reliable, accurate and precise. The monoclonal ELISA is simpler, quicker and easier to perform in routine use. Data generated is directly comparable to that generated by PEG precipitation. This methodology would be suitable for laboratories currently measuring Protein S by ELISA. PMID- 9218151 TI - Expression of von Willebrand factor, P-selectin (CD62P) and thrombomodulin in human endothelial cells in culture modulated by cyclosporin A. AB - Endothelial cells were isolated from human umbilical veins and from saphenous veins and were cultured in the presence and absence of cyclosporin A (CSA). Cell morphology and growth characteristics were monitored continuously by video recording microscopy. The cells and culture supernatants were analysed for von Willebrand Factor (VWF), P-Selectin (CD62P) and thrombomodulin (TM) by immunocytochemistry and immunoassay. CSA had little effect on confluent cells in culture. In contrast, the characteristics of both umbilical vein and saphenous vein cells were markedly altered when CSA was added to subconfluent cultures. Under these conditions, CSA did not appear to be directly cytotoxic at the concentrations used but marked changes in cell morphology including the appearance of large multinucleate forms were recorded. Increased amounts of VWF and TM were detected in the culture supernatants of cells grown continuously in the presence of CSA and correspondingly depressed expression of cell associated antigens was observed. Heterogeneous and depressed distribution of CD62P was seen after 5-6 days culture but cellular expression of this ligand did not appear to be affected specifically by the presence of CSA. In addition, mechanically disrupted confluent endothelial cells failed to recover in the presence of CSA as effectively as similar cells grown in the absence of CSA. The results showed that CSA disturbed actively growing cells more severely than quiescent cells and might help to clarify the role of CSA in vascular disturbances in vivo. PMID- 9218152 TI - Insulin receptor substrate-1 gene polymorphism and cardiovascular risk in non insulin dependent diabetes mellitus and patients undergoing coronary angiography. AB - Clustering of risk factors for cardiovascular disease related to insulin resistance may account for the increased incidence of vascular disease in these conditions and in non-diabetic subjects. To investigate the relationship between a coding polymorphism in the insulin receptor substrate-1 gene and the presence of cardiovascular risk factors, 209 patients with NIDDM and 452 subjects investigated for coronary artery disease (CAD) were studied. In the NIDDM subjects 22 (10.5%) were heterozygous at codon 972 for a polymorphism which codes for a glycine to arginine substitution and 187 (89.5%) were homozygous for the wild type. Patients with the mutation had lower levels of cholesterol compared with wild type (mean, 95% confidence intervals), 5.3 (4.9-5.8) vs 6.0 (5.9-6.2) mmol/l, respectively (P = 0.002), triglyceride 1.7 (1.4-2.1) vs 2.2 (2.0-2.4) mmol/l (P = 0.051), factor VII:C activity 109.5 (85.5-133.5) vs 133.5 (127-140)% (P = 0.057) and PAI-1 antigen, 16.0 (10.5-24.3) vs 22.2 (20.0-24.6) ng/ml (P = 0.054). There were no differences in body mass index, indices of glycaemic control, fasting insulin or the prevalence of hypertension. In patients with CAD, 55 (12.7%) were carriers of the mutation (including three homozygotes) (NIDDM vs CAD, NS). Although similar trends in cholesterol, factor VII, PAI-1 antigen and triglyceride existed between carriers of the mutation and the wild type, none reached statistical significance. The results indicate that the IRS-1 gene is not implicated in the pathogenesis of NIDDM or CAD. PMID- 9218153 TI - An EIA method on single donor solubilized HLA antigens for the identification of anti-HLA antibodies. AB - An enzyme immunoassay (EIA) method based on solubilized human leucocyte antigens (HLA) derived from single donor platelets is described. The EIA results on these solubilized single donor HLA antigens (SDszHLA) correlated well with the complement dependent cytotoxicity (CDC) results on the lymphocytes of the same donors and also with the panel reactivity (PRA) in CDC. A concordancy rate of 78% was found for individual HLA specificities. The EIA+/CDC- ('false positive') discrepancies were more pronounced than EIA-/CDC+ ('false negative') discrepancies and varied for the different donors. To confirm discrepancies, our method was compared with a commercial PRA-STAT EIA method (based on secreted soluble HLA antigens). The same discrepancies between CDC and PRA-STAT EIA were found and are probably due to the higher and different sensitivity (e.g. non complement fixing antibodies) of EIA methods. A SDszHLA EIA method allows the identification of HLA specificities of HLA-antisera. The possibility of using individual and selected donors for the production of SDszHLA allows the directed search for well defined HLA specificities in order to confirm anti-HLA specificities found in other anti-HLA screening methods. An individualized HLA panel can be established with the support of blood banks that have HLA typed blood and platelet donors. PMID- 9218154 TI - Measurement of feto-maternal haemorrhage: a comparative study of three Kleihauer techniques and tow flow cytometry methods. AB - In the UK a Kleihauer test is routinely performed on all RhD-negative women after the birth of an RhD-positive child to ensure that an adequate dose of anti-D immunoglobulin is given. The results of Kleihauer testing are interpreted to assess the volume of any feto-maternal haemorrhage and additional anti-D immunoglobulin is administered if necessary. A similar procedure is followed ante natally when incidents occur known to be associated with alloimmunization. The performance of Kleihauer tests is difficult to standardize and there can be problems in interpreting the volume of feto-maternal haemorrhage. The use of flow cytometry to measure feto-maternal haemorrhage is reported to give more accurate and reliable results. This study compared three different Kleihauer methods and two different flow cytometry techniques all performed using the same maternal sample and within a single laboratory. The results demonstrated variability between the Kleihauer tests used and also in the flow cytometry measurements. A well-performed Kleihauer test would still appear to be useful as a screening technique for detection of feto-maternal haemorrhage. However, accurate quantitation of size of feto-maternal haemorrhage is more reliably determined by flow cytometry. PMID- 9218155 TI - The Rhnull phenotype in an English individual: haematological, serological and immunological studies. AB - We have characterized the first case of Rhnull phenotype to be identified in England. The red cells were serologically negative for C, c, Cw, D, E, e, f, hr B, Rh17, Lw a, Lw ab and Duclos, while the patient's serum contained anti-Rh29, which was subsequently boosted by transfusion. The Rh phenotype of the patient's son (R1r) confirmed that this was a regulator type of Rhnull in the patient. Follow up studies confirmed the presence of a mild chronic anaemia with stomatocytes and spherocytes; electron microscopy revealed the presence of cells with deep central indentations. Osmotic fragility was increased to a level intermediate between normal and hereditary spherocytic controls. The presence of ongoing haemolysis was indicated by a mild reticulocytosis and splenomegaly. The potent anti-Rh29 has made the provision of compatible blood difficult and autologous units have been frozen. The case illustrates the rare phenomenon of the Rhnull phenotype which not only causes problems for the transfusionist but should also be recognized as a cause of haemolytic anaemia secondary to a membrane defect. Blood film and Rh phenotyping are useful preliminary investigations in suspected cases. PMID- 9218156 TI - The place of myeloid growth factors in the treatment of hairy-cell leukaemia. AB - A patient with hairy-cell leukaemia was treated with granulocyte colony stimulating factor lenograstim (Granocyte) 300 micrograms daily by subcutaneous injections. His pre-existing neutropenia remitted and the therapy was continued for a total of 4 months. When the therapy was discontinued the neutropenia returned. There was no evidence that the growth factor itself had any disease modifying activity. PMID- 9218157 TI - Myelodysplasia occurring after fludarabine treatment for chronic lymphocytic leukaemia. AB - The purine analogue, fludarabine, is of proven efficacy in the treatment of lymphoid malignancies. The drug appears to be well tolerated with minimal side effects, and few toxicities have been observed. A case of myelodysplasia occurring after therapy with fludarabine is presented and its implications are discussed. PMID- 9218158 TI - Ultrastructure of myeloma cells producing parathyroid hormone-related peptide. AB - We examined the ultrastructure of myeloma cells producing parathyroid hormone related peptide. The nucleus was mature and the cytoplasm was well-developed, being classified into the mature type with slight nucleo-cytoplasmic asynchrony. Although various nuclear and cytoplasmic abnormalities commonly observed in myeloma cells were recognized, a multilamellar body-like structure which has not been reported previously in myeloma cells was characteristically observed. Numerous nuclear and cytoplasmic abnormalities observed in the myeloma cells of this case were considered to have resulted from increased and aberrant proliferation of myeloma cells. We reported previously that the immature nucleus and various nuclear and cytoplasmic abnormalities are related to poor prognosis. Thus, the ultrastructural findings of myeloma cells in this case is not inconsistent with the short survival time. PMID- 9218159 TI - The use of low dose Orgaran in heparin-induced thrombocytopenia associated with in vitro platelet aggregation at higher Orgaran concentrations. AB - We report a case of heparin-induced thrombocytopenia with in vitro antibody cross reactivity by platelet aggregometry to both low molecular weight heparin and the heparinoid Org 10172 (Orgaran). The in vitro reactivity with Orgaran was only present at the upper limit of concentrations that would normally be used therapeutically. Low dose Orgaran therapy was initiated, allowing successful renal replacement therapy without invoking further thrombocytopenia or thrombosis. Interestingly, in vitro platelet aggregometry following treatment did not reveal increasing sensitivity to Orgaran. This case indicates that negative in vitro platelet aggregometry at defined lower concentrations of Orgaran may predict in vivo safety at the same levels despite positive platelet aggregometry reactions at higher concentrations of Orgaran. PMID- 9218160 TI - Uniformity of anticoagulation for full blood counting. PMID- 9218161 TI - What are the therapeutic needs in schizophrenia and how are they satisfied by new antipsychotics? AB - Although considerable progress has been made in the management of schizophrenia, much remains to be achieved in meeting the therapeutic needs of patients with this illness. Conventional antipsychotics have revolutionized treatment, but a substantial proportion of patients derive inadequate benefit and many suffer from adverse effects. Compliance with medication remains an enormous problem. With a number of new drugs soon to be available, it is hoped that valuable alternatives can be provided, increasing the possibility of good therapeutic response and increasing the benefit-to-risk ratio. PMID- 9218162 TI - Improvement of negative symptoms: Concepts, definition and assessment. AB - The development of new medications for the treatment of schizophrenia is closely tied to the concept of the disorder and its characteristic symptoms. In recent years, the symptoms have been divided into two broad categories: positive and negative. Positive symptoms tend to command clinical attention and to be treatment-responsive, which negative symptoms are more insidious and disabling, but less spectacular. Negative symptoms, which are similar to the core symptoms of schizophrenia defined by Krapaelin and Bleuler, have not received much attention until recently because of concern about reliability. However, rating scales with good reliability are now available for use in clinical and neurobiological studies. Clinical drug trials are also meeting the challenge of documenting the response to carefully rated negative symptoms. In addition, they are exploring the effects of medication on negative symptoms that are primary rather than secondary to reduced extrapyramidal side effects, or of medication that lowers levels of depression or decreases the demoralizing effects of positive symptoms. One ideal strategy for identifying the effects of medication on primary negative symptoms is to study patients with high levels of negative symptoms and low levels of positive symptoms, who are in the early stages of the illness and have been given minimal treatment with classical neuroleptics. Such samples are highly informative, but difficult to collect. Alternatively, when all confounders cannot be eliminated, the effects of medication can be explored using statistical techniques to examine covariance. PMID- 9218163 TI - Amisulpride compared with standard neuroleptics in acute exacerbations of schizophrenia: three efficacy studies. AB - Three double-blind studies that included acutely ill schizophrenic patients were undertaken to assess the efficacy of amisulpride, an atypical antipsychotic with a selective affinity for dopamine D2, and D3, receptors. In the first study fixed doses of amisulpride (400, 800 and 1200 mg/day) and 16 mg/day of haloperidol were compared with 100 mg/day of amisulpride for a 4-week period. Efficacy was assessed using the Brief Psychiatric Rating Scale (BPRS), subscales of the Positive and Negative Symptom Scales (PANSS) and the Clinical Global Evaluation (CGI). Data were obtained on 319 subjects with the revised third-edition Diagnostic and Statistical Manual of Mental Disorders (DSM-III-R) criteria for schizophrenia. The highest improvement, in terms of BPRS total scores, occurred in the two groups taking 400 mg or 800 mg amisulpride. These doses also induced less extrapyramidal disturbance compared with haloperidol. The second study compared amisulpride with haloperidol: 95 patients were treated for 6 weeks with 800 mg/day of amisulpride and 96 patients with 20 mg/day of haloperidol. Efficacy criteria were the same as in the first study. Amisulpride at 800 mg/day was at least as efficacious as 20 mg haloperidol for the positive symptoms of schizophrenia, but was significantly more effective against negative symptoms (PANSS negative subscale, P = 0.038). There was also a significantly lower incidence of parkisonism in the amisulpride group. The third study compared amisulpride with flupenthixol: 70 patients were treated with 1000 mg/day of amisulpride and 62 patients with 25 mg/day of flupenthixol for a 6-week period. The mean improvement in the BPRS total score was greater in the amisulpride group, although this difference just failed to reach statistical significance (P = 0.059). Both drugs improved negative symptoms (Scale for the Assessment of Negative Symptoms), but the effect was more marked with amisulpride. Differences were statistically different in favour of amisulpride for positive symptoms. These studies show that amisulpride has optimal efficacy at doses of between 400 and 800 mg/day. It was at least as effective as reference drugs in the treatment of positive symptoms, and also reduced negative symptoms. Furthermore, amisulpride provoked fewer extrapyramidal side effects than standard reference drugs. PMID- 9218164 TI - Clinical update on amisulpride in deficit schizophrenia. AB - Amisulpride is an atypical antipsychotic drug. Low doses increase dopaminergic transmission via presynaptic blockade and are effective in patients with predominantly negative (deficit) symptoms of schizophrenia. In three double-blind studies, two short-term and one medium/long-term, comparing amisulpride with placebo, 272 patients with schizophrenia were carefully selected for a predominance of negative symptoms (low severity of positive symptoms, depression and extrapyramidal side effects). Fixed daily doses of amisulpride (100 and 300 mg) were used in a 6-week dose-range finding study (104 patients). Daily doses of between 50 and 100 mg amisulpride were used in a second 6-week study (27 patients), and in a third study, 100 mg amisulpride was administered for 6 months (141 patients) with an extension of up to 1 year. Mean total scores for the Scale for the Assessment of Negative Symptoms (SANS) at baseline in the different treatment groups varied from 98 to 74, and improved significantly in the amisulpride groups (mean change from 24 to 40 points) compared with the placebo groups. Positive symptoms measured by the Scale for the Assessment of Positive Symptoms (SAPS) were low at baseline, and the change at the end of the studies was minimal. Extrapyramidal side effects were of low severity in these studies and parkinsonism scores for amisulpride were not different from placebo. Overall, these findings indicate that the improvement in negative symptoms was not linked to a concomitant improvement in positive symptoms, parkinsonism or depressive symptoms as would be expected in the case of secondary negative symptoms. The results of these studies thus confirm the efficacy of amisulpride in schizophrenic patients with primary negative or deficit symptoms at a dose of 100 mg/day. PMID- 9218165 TI - Amisulpride: from animal pharmacology to therapeutic action. AB - Amisulpride is a benzamide derivative with a unique neurochemical and psychopharmacological profile. This compound has selective affinity for human dopamine D3 and D2 receptor subtypes in vitro (binding constant, K approximately 3 nmol/l) and blocks functional responses mediated by these receptors. In ex vivo binding studies, amisulpride is twice as selective for D3 as for D2 receptors. At low doses, it preferentially blocks presynaptic dopamine autoreceptors (increase in dopamine release in vivo in the rat olfactory tubercle, 50% effective dose, ED50 3.7 mg/kg), while postsynaptic dopamine receptor antagonism is apparent at higher doses (decrease in striatal acetylcholine levels, ED50 approximately 60 mg/kg). Anisulpride preferentially stimulates dopamine synthesis and displaces 3H raclopride binding in vivo in the limbic system rather than the striatum. It antagonizes apomorphine-induced hypothermia in mice and amphetamine-induced hypermotility in rats at low doses (ED50 2-3 mg/kg), blocks apomorphine-induced climbing and spontaneous grooming in mice, blocks apomorphine-induced gnawing in rats at higher doses (ED50 19-115 mg/kg) and does not induce catalepsy at 100 mg/kg. The preferential antagonism by amisulpride of presynaptic D2/D3 receptors is reflected behaviourally in the potent blockade of apomorphine-induced effects mediated by dopamine autoreceptors (yawning and hypomotility: ED50 0.2 and 0.3 mg/kg, respectively) compared with those medicated by postsynaptic D2 receptors (e.g. gnawing: ED50 115 mg/kg). Moreover, low doses of amisulpride induce prohedonic (potentiation of food-induced place preference) effects in rats. The atypical neurochemical and psychopharmacological profiles of amisulpride may explain its therapeutic efficacy on both positive and negative symptoms of schizophrenia. PMID- 9218166 TI - Conformation of nucleoplasmin and its interaction with DNA-protamine complex as a simple model of fish sperm nuclei. AB - Nucleoplasmin was isolated from Xenopus laevis eggs and purified by an improved method using an open column. Its conformation was investigated spectrophotometrically by UV, CD and fluorescence. It was shown that alpha-helix content of nucleoplasmin was 30-40%, and one of the two tryptophan residues in nucleoplasmin located in the hydrophobic surroundings and the other in the relatively hydrophilic surroundings. The isolated nucleoplasmin was found to decondense sperm nuclei of salmon also, suggesting a possibility of the existence of nucleoplasmin-like protein in fish as well. Collapse of the protamine (salmine)-DNA complex as a simple model for fish sperm nuclei by nucleoplasmin was directly observed by measuring OD320 of aqueous protamine-DNA mixtures. This is a molecular level observation for the removal of protamine from DNA-protamine complex. PMID- 9218167 TI - Associative behaviour of hydrophobically modified carboxymethylpullulan derivatives. AB - Hydrophobically modified carboxymethylpullulan (HMCMP) samples were obtained by reaction of small amounts of C16 alkylamine on carboxylic groups of the corresponding polyacid. The molar contents of alkyl chains ranged from 1.3 to 6.8% with respect to the anhydroglucose units (AGU) and the degree of substitution (DS) of carboxylic groups varied from 0.76 to 0.84. Solution properties of the sodium salt of HMCMPs were studied mainly by viscometric and size-exclusion chromatography/light-scattering methods. The low-shear viscosity of modified pullulans in 0.1 M NaCl solutions drastically increases with the hydrophobic content and polymer concentrations and the 6.8% modified sample has a quite pseudoplastic behaviour. These data showed that the polymers aggregated intermolecularly and displayed a compact globular structure in dilute solution. Furthermore, addition of NaCl or ethanol induced a decrease in viscosity although the molecular weights remained approximately constant. These results are consistent with a collapse of the polysaccharide aggregates. PMID- 9218169 TI - The effects of ageing of amylopectin gels on the diffusion of BSA. AB - Amylopectin gels, aged for different times (0-39 days), were used to investigate the effects of ageing on the diffusion of bovine serum albumin. It was found that the rate of the diffusion initially decreased rapidly as the ageing time increased (< or = 10 days). Thereafter, it continued to decrease but more slowly. PMID- 9218168 TI - DSC studies on bovine serum albumin denaturation. Effects of ionic strength and SDS concentration. AB - This work analyzed the thermal denaturation process of defatted bovine serum albumin (BSA). DSC measurements were performed on changing the pH, the ionic strength and the sodium dodecyl sulfate (SDS) concentration. These data have been compared with those previously obtained by us and other authors. The purpose of these measurements was to study the correlation between the three-dimensional organization of BSA native protein structure and its thermodynamic stability and to clarify the non-covalent interactions between the globular proteins and amphipathic molecules. These measurements have shown that the thermal denaturation is always irreversible regardless of pH, ionic strength and SDS concentration. The nature of the irreversible process superimposed on the protein unfolding is discussed. The strong stabilizing effect of NaCl on the BSA native structure has been found for the range 0-1.0 M. It is worth noting that the calorimetric curves, confined to the pH region studied, could not be represented by a two-state transition model; they were deconvoluted as the sum of two independent two-state transitions. These transitions were correlated to the domain structure of BSA. Sodium dodecyl sulfate has a net stabilizing effect up to a molar ratio of 10:1 (ligand to protein). In this range of concentrations the presence of SDS cause a biphasic profile of excess heat capacity. A simple thermodynamic model was developed in attempt to reproduce the experimental DSC profiles and collect information regarding the binding equilibrium of SDS. PMID- 9218170 TI - A cyclic disulfide peptide reproduces in solution the main structural features of a native antigenic site of foot-and-mouth disease virus. AB - A cyclic disulfide peptide corresponding to the G-H loop sequence 134-155 [replacement Tyr136 and Arg153 with Cys] of the capsid protein VP1 of foot-and mouth disease virus (FMDV) isolate C-S8c1 was examined by proton 2D-NMR spectroscopy in water and in 25% HFIP/water. In water, NMR data supported the presence of a non-canonical turn in the central, conserved cell adhesion RGD motif and suggested the presence of a nascent helix in the C-terminal part, stabilized and slightly extended upon addition of 25% HFIP, a secondary structure stabilizing cosolvent. The formation of the C-terminal helix was evidenced by combined analysis of NOE connectivities, H alpha chemical shifts, 3JNH-H alpha coupling constants and amide temperature coefficients. Surprisingly, these global structural features of the cyclic peptide in solution show similarities to previous X-ray structure analysis of (a) a shortened linear peptide complexed with a antivirus antibody and (b) the G-H loop represented on the chemical reduced viral surface of a different serotype. Thus, even in entirely different biological environments the cyclic peptide reflect similar structural features, reinforcing the concept that this viral loop behaves as an independent structural and functional unit. PMID- 9218171 TI - Influence of substituent of direct dye having bisphenylenebis(azo) skeletal structure on structure of nascent cellulose produced by Acetobacter xylinum [I]: different influence of direct red 28, blue 1 and 15 on nascent structure. AB - The difference of influence of a certain kind of direct dye on the structure of nascent microbial cellulose was examined, with Direct Red 28 have a biphenylenebis(azo) skeletal structure; Direct Blue 1 having two hydroxyl, two methoxy and two sulfonate groups more than Direct Red 28; and Direct Blue 15 whose sulfonate groups position are different compared to Direct Blue 1. It became clear that the product in the presence of a direct dye (in particular, Direct Red 28) has the structure in which the dye molecule is included between the monolayer in the cellulose sheets corresponding to the (110) plane of microbial cellulose. On the other hand, the structure of the product in the presence of Direct Blue 1 and 15 contains conceived cellulose II structure which occurred due to be removal of dye during the rinsing process as a result of larger hydrophilicity than its affinity toward cellulose. Solid state 13C NMR and deuteration-IR measurements showed that the product in the presence of direct dye is in a noncrystalline state, although X-ray measurements indicated that they are in a crystalline state. These results support the inclusion of a dye between the (110) planes. Solid state 13C NMR and deuteration-IR reveal that the crystal structure of cellulose regenerated from the product in the presence of Direct Red 28 is similar to cellulose IVI, while that from each Direct Blue 1 and 15 product is cellulose II. The difference of the influence of the former and the latter on the nascent cellulose seemed to be caused mainly by the number of sulfonate groups, although the influence of hydroxyl and methoxy groups is not clear at present. PMID- 9218172 TI - Effect of molecular weight and urea on the conformation of chitosan molecules in dilute solutions. AB - The effects of molecular weight and urea on the magnitude of the Mark-Houwink exponent a and the relative stiffness parameter B of chitosan molecules in dilute solutions were analyzed. The results show that in solutions with ionic strengths between 0.01 and 0.3 M, the relative chain stiffness parameter B and the Mark Houwink exponent a of chitosans whose molecular weights were between 22.3 x 10(4) and 91.4 x 10(4) fell between 0.143 and 0.152 and from 0.404 to 0.497, respectively; whereas for chitosans whose molecular weights were between 7.8 x 10(4) and 14.8 x 10(4) these values fell between 0.110 and 0.138 and from 0.653 to 1.009, respectively. Both results indicate that the stiffness and conformations of small molecular weight chitosans were more stiff and extended, respectively, than higher molecular weight ones, and that molecular weight induced conformational transition occurred. Chitosans in solutions containing 4 M urea possessed a rod-shaped conformation in both molecular weight domains, and no molecular weight-induced conformational transitions occurred. PMID- 9218173 TI - Marked effect of DNA on collagen fibrillogenesis in vitro. AB - Growth of collagen fibrils in the presence of DNA was more rapid than that in the absence of DNA. Some of collagen fibrils formed in the presence of DNA were significantly wider than those in the absence of DNA. Moreover, the cross bandings were also very distinct in spite of using pepsin-digested collagen. These results suggest that DNA not only adsorbs to collagen but induces the extraordinary fibrillogenesis of collagen. PMID- 9218174 TI - Exercise and blood pressure: Walk, run or swim? PMID- 9218175 TI - How to assess vascular remodelling in small and medium-sized muscular arteries in humans. AB - The study of vascular wall changes in humans has generated great interest with the increasing realization that, independently of the potential contribution to mechanisms involved in blood pressure elevation, these structural alterations (remodelling) or functional changes may contribute to the complications of elevated blood pressure. Moreover, some of these changes may be corrected partially or totally by administration of antihypertensive agents and other drugs. This has fuelled interest in the techniques used to evaluate changes in the vascular wall in humans, which are reviewed critically here with a focus on human studies in hypertension. Remodelling of large and small arteries has different characteristics, and is studies with different techniques. In hypertensive patients, small arteries less than 400 microns in diameter exhibit a reduction in lumen diameter, accompanied sometimes but not always by an increase in media width or in media cross-section. The study of capillaries and small arteries of the skin or the eye can be performed non-invasively, but for the sake of obtaining the information of interest in hypertension, at present invasive techniques are required to investigate small arteries. These consist of a biopsy of subcutaneous tissue, usually from the gluteal region, and the study of vessels after they have been mounted on a 'wire myograph' or on a pressurized system. In contrast to small arteries, large arteries from hypertensive humans present increases in media width without a significant reduction in the lumen diameter (when studied under conditions isobaric relative to those in normotensive subjects). Conduit arteries may be studies non-invasively with the use of ultrasound techniques. The study of large elastic arteries is not addressed here. The use of echo-tracking devices to study muscular medium-sized arteries such as the radial artery is described. The relative advantages and disadvantages of these techniques, the questions which may be asked and the relevance of the information obtained using these approaches are discussed. PMID- 9218176 TI - Is the 'clinic-home blood pressure difference' associated with psychological distress? A primary care-based study. AB - OBJECTIVE: To determine whether there is an association between the 'clinic-home blood pressure difference' (CHBPD) and psychological distress in a sample not selected without regard to blood pressure and hypertension status. DESIGN: A cross-sectional study. SETTING: An academic family medicine department in Toronto, Canada. PARTICIPANTS: Consecutive attenders (n = 214) of the primary care facility. Subjects aged less than 16 years and those being administered psychotropic or blood pressure-lowering agents were excluded. MAIN OUTCOME MEASURES: The CHBPD was calculated from clinic blood pressure readings and self measurements by subjects at home; psychological distress was measured by the 30 item version of the General Health Questionnaire (GHQ). RESULTS: No significant association between the CHBPD and psychological distress could be shown for systolic and diastolic blood pressures. The same applied to GHQ subdomains and the CHBPD modelled on several independent variables by multiple linear regression analyses. CONCLUSION: The results from this study, using a large sample drawn from a community, support the view that the CHBPD is not related to anxiety, depression and other forms of psychological distress, but rather is a reaction specific to the clinic setting itself. PMID- 9218177 TI - Sex differences in the pharmacological treatment of hypertension: a review of population-based studies. AB - OBJECTIVE: To summarize all available literature on sex differences in the pharmacological treatment of hypertension with respect to the percentage of hypertensive patients treated pharmacologically and the selection of antihypertensive drugs. The influences of the calendar period, age, definition of hypertension, prevalence of hypertension and country on these sex differences were examined. DATA IDENTIFICATION: A secondary analysis of data from 46 population-based studies in 22 countries on the prevalence of pharmacologically treated hypertension was conducted to estimate sex ratios for the prevalence of drug treatment for hypertension. RESULT: Overall, women with hypertension were 1.33-fold [95% confidence interval (CI) 1.32-1.34] more likely to be treated pharmacologically for hypertension than were hypertensive men. With increasing age, the female: male ratio for pharmacological treatment of hypertension decreased from 2.26 (95% CI 1.56-3.27) at ages 20-29 years to 1.22 (95% CI 1.11 1.34) at ages 60-69 years. In all countries more women than men were treated for hypertension, with the biggest difference observed in the USSR (1983-1986), where about twice as many women as men were treated for hypertension. Women more frequently used diuretics, whereas men more often used beta-blockers, angiotensin converting enzyme inhibitors and calcium antagonists. CONCLUSIONS: Hypertensive women are more often treated for hypertension than hypertensive men and their pattern of use of antihypertensive drugs differs from that of men. Further research is required in order to explain sex differences in the treatment of hypertension with respect to the prevalence of pharmacological treatment of hypertension and choice of antihypertensive drugs, and to investigate the consequences of this difference for long-term outcomes. PMID- 9218178 TI - Genetic studies of the renin-angiotensin system in arterial hypertension associated with non-insulin-dependent diabetes mellitus. AB - OBJECTIVE: To determine whether angiotensinogen (AGT) and angiotensin II type 1 (AT1) receptor genes contribute to the development of arterial hypertension in members of French Caucasian families and in subjects with hypertension associated with non-insulin-dependent diabetes mellitus (NIDDM). METHODS: Sibpair linkage analyses were performed with microsatellites near the AGT and AT1 receptor genes in 179 hypertensive sibpairs from 69 NIDDM kindreds. In addition, population/association studies were performed with the M235T and T174M polymorphisms of the AGT gene, and the A1166C polymorphism of the AT1 receptor gene. RESULTS: No evidence for linkage between the AGT and AT1 receptor loci and hypertension was observed. In addition, the distributions of genotypes of AGT and AT1 receptor gene polymorphisms did not differ significantly among a group of unrelated individuals with both hypertension and NIDDM (n = 188) and three groups of unrelated control subjects with NIDDM (n = 117), hypertension (n = 75) or none of these conditions (n = 125). CONCLUSIONS: These results suggest that the AGT and AT1 receptor genes are not major genetic determinants of hypertension associated with NIDDM in this population, although we can not exclude the possibility that these loci make a minor contribution in a polygenic context. PMID- 9218179 TI - Molecular variant M235T of the angiotensinogen gene is associated with essential hypertension in Taiwanese. AB - OBJECTIVE: To examine the association of the molecular variants of the angiotensinogen (AGT) gene with essential hypertension in Taiwanese. METHODS: We conducted a case-control study concerning 151 subjects, 102 hypertensives and 49 normotensives. We created a rapid mini-sequencing method based on dye-terminator cycle sequencing to simultaneously detect the M235T and T174M variants of the AGT gene for each subject. RESULTS: The genotype and allele distribution of the M235T variant differed significantly in hypertensives and normotensives (chi 2 = 11.106, P = 0.004 and chi 2 = 6.453, P = 0.011, respectively), whereas those of the T174M variant did not differ (chi 2 = 0.004, P = 0.998 and chi 2 = 0.032, P = 0.858, respectively). The odds ratio for hypertension was 3.64 (95% confidence interval 1.56-8.49) for subjects with the C/C genotype of the M235T variant compared with other genotypes of 2.87 (95% confidence interval 1.76-4.68) for those carrying allele C versus those carrying allele T. CONCLUSION: The molecular variant M235T, but not T174M, of the AGT gene is associated significantly with essential hypertension in this Taiwanese population. The genotype C/C or allele C is a risk factor for hypertension. The underlying mechanism of this association needs to be elucidated further. PMID- 9218181 TI - Structural changes of small resistance arteries in spontaneously hypertensive rats after treatment with various doses of lacidipine. AB - OBJECTIVE: To evaluate the modifications of the morphology of mesenteric small resistance vessels in spontaneously hypertensive rats (SHR) induced by lacidipine treatment. METHODS: Lacidipine was administered at three different dosages, 20, 10, and 0.3 mg/kg per day. Fifty rats were studied. Nine SHR and 11 Wistar-Kyoto (WKY) rats were not treated. Each lacidipine dose was administered to 10 SHR. The drug and the placebo were administered by gavage from age 4 to age 12 weeks. The blood pressure was measured noninvasively every week. The animals were killed when they were aged 13 weeks, and the relative left ventricular mass (left ventricular weight plus septum weight/body weight) was calculated. Small mesenteric resistance vessels were dissected and mounted on a micromyograph (Mulvany's technique), and morphological parameters of the vessels were studied (media thickness and media: lumen ratio). RESULTS: The systolic blood pressure of SHR administered 20 and 10 mg/kg lacidipine per day was reduced significantly during the treatment period, whereas that of rats treated with 0.3 mg/kg lacidipine per day did not change. A significant reduction in media: lumen ratio was observed for all three groups of treated rats, including those to which 0.3 mg/kg lacidipine per day had been administered, and no reduction in systolic blood pressure could be detected. The relative left ventricular mass was reduced significantly only in rats to which 20 and 10 mg/kg lacidipine per day had been administered. CONCLUSION: A significant reduction in magnitude of vascular structural alternations was observed even in SHR treated with a low, nonhypotensive dose of lacidipine. PMID- 9218180 TI - Endothelial dysfunction in spontaneously hypertensive rats: consequences of chronic treatment with losartan or captopril. AB - BACKGROUND: Hypertension is associated with endothelial dysfunction characterized by decreased endothelium-dependent relaxations and increased endothelium dependent contractions. Angiotensin converting enzyme inhibitors and thromboxane A2 receptor antagonists decreased the endothelium dysfunction in hypertensive animals. OBJECTIVE: To investigate the effects of prolonged treatment with losartan on endothelium-dependent and -independent relaxations and contractions in aortic rings from spontaneously hypertensive rats (SHR). MATERIAL AND METHODS: Male SHR aged 16 weeks were treated for 12 consecutive weeks either with 10 mg/kg losartan per day or with 60 mg/kg captopril per day administered via their drinking water. The systolic blood pressure was evaluated basally and during week 12. At the end of the treatment period, the vascular reactivity in aortic rings was studies. A group of rats treated with captopril was studies as a reference group. RESULTS: Losartan and captopril reduced the blood pressure significantly and comparably. Both drugs enhanced acetylcholine-induced relaxations and reduced the maximal contractile response to acetylcholine in the presence of NG-nitro-L arginine methyl ester (L-NAME). Contractile responses to phenylephrine, endothelin-l and U46619 were not affected by these treatments. Increased relaxing responses to superoxide dismutase were observed only in captopril-treated rats. Losartan reduced the contractile response to angiotensin II. By contrast this contractile response was elevated in rats treated with captopril. CONCLUSIONS: Prolonged antihypertensive treatments with losartan and captopril decreased the endothelial dysfunction in aortic rings from SHR not only by enhancing NO dependent relaxations but also by reducing the contractions in response to an endothelium-derived contracting factor. The results further confirm that an endothelium-derived contracting factor plays a role in vascular dysfunction in SHR and the relationships between this factor and angiotensin II. PMID- 9218182 TI - Arginine vasopressin inhibits interleukin-1 beta-stimulated nitric oxide and cyclic guanosine monophosphate production via the V1 receptor in cultured rat vascular smooth muscle cells. AB - BACKGROUND: It has been reported that various vasoactive substance modulate cytokine stimulated nitric oxide (NO) production in many cell types. OBJECTIVE: To examine the effects of arginine vasopressin (AVP) on the production of NO and cyclic GMP (cGMP), and on inducible nitric oxide synthase (INOS) in cultured rat vascular smooth muscle cells (VSMC). DESIGN: Because VSMC possess the V1 receptor which clauses vascular contraction and respond to various cytokines for producing NO, we used rat VSMC and selected interleukin-1 beta (IL-1 beta) as a potent stimulator of NO production among various cytokines. We also measured cGMP production, which is the final mediator of NO-induced vascular relaxation, in order to evaluate the physiologic meaning of the present study. METHODS: VSMC were incubated with test agents for 24 h except for a time-course study. Nitrite as a stable end product of NO was measured in the medium. Intracellular cGMP contents were assayed by enzyme immunoassay. INOS messenger RNA expression was analyzed by Northern blotting. RESULTS: AVP inhibited IL-1 beta-induced nitrite production in a dose- and time-dependent manner with concomitant changes in intracellular cGMP contents. On the other hand, AVP did not affect nitrite and cGMP production in the absence of IL-1 beta. Inhibition of nitrite and cGMP production by AVP was reversed by administration of the specific V1 receptor antagonist [1-(beta-mercapto-beta,beta- cyclopentamethylene propionic acid), 2-(O methyl)-tyrosine] -Arg8-vasopressin) but not by the oxytocin (OXT) receptor antagonist [d(CH2(5)), TyrMe2, Orn8]-Vasotocin. Administration of the V1 receptor antagonist or OXT receptor antagonist alone did not affect IL-1 beta-stimulated nitrite and cGMP production. Although administration of AVP inhibited IL-1 beta induced INOS messenger RNA expression, administration of the V1 receptor antagonist but not of the OXT receptor antagonist reversed this inhibition. CONCLUSION: It is suggested that AVP inhibits IL-1 beta-induced NO and cGMP production via the V1 receptor but not via the OXT receptor in VSMC. AVP can cause vascular contraction not only through direct action but also through indirect action by inhibiting NO production under some inflammatory conditions. PMID- 9218183 TI - Role of kinins and angiotensin II in the vasodilating action of angiotensin converting enzyme inhibition in rat renal vessels. AB - OBJECTIVE: To assess directly the vasodilating effects of angiotensin converting enzyme (ACE) inhibition in different renal vessels and to determine the role of kinins and angiotensin II (ANGII) therein. METHODS: Lumen diameters of different vessels and glomerular blood flows were measured in cortical and juxtamedullary glomeruli by in-vivo microscopy in the split hydronephrotic kidney of anesthetized female Wistar rats. RESULTS: Injection of the ACE inhibitor quinapril at a dose of 0.9 mg/kg intravenously, which blocks conversion of locally applied angiotensin I (1 mumol/l), increased glomerular blood flows by 39 +/- 6 and 18 +/- 4% in cortical and juxtamedullary glomeruli, respectively, due to vasodilatation in all renal vessels. The most pronounced vasodilatation was observed in interlobular arteries (19 +/- 2%) and in cortical afferent arterioles (16 +/- 3%). Pretreatment of the hydronephrotic kidney by local application of 40 nmol/l Hoe140, a bradykinin B2 receptor antagonist, or 3 mumol/l valsartan, an ANGII type 1 receptor antagonist, attenuated the vasodilatation in response to quinapril. ANGII receptor blockade affected only weakly, whereas bradykinin receptor blockade blunted markedly, the quinapril-induced vasodilatation, suggesting that kinins play an important role in our experimental model. Administration of valsartan, which abrogated the renal vasoconstriction induced by 10 nmol/l ANGII completely, caused vasodilation of magnitude similar to that caused by administration of quinapril. Yet, the vasodilatation induced by the combination of valsartan and quinapril was significantly larger than that induced by administration of quinapril alone in interlobular arteries, afferent arterioles, and cortical efferent arterioles. CONCLUSIONS: Our results indicate that kinins and ANGII can contribute to the renal vasodilatation in response to ACE inhibitors, but ACE inhibitors appear to have only minor effects on ANGII levels in those renal vessels, which are the well-known sites of renin expression. PMID- 9218184 TI - Elevated vascular angiotensin converting enzyme mediates increased neointima formation after balloon injury in spontaneously hypertensive rats. AB - OBJECTIVE: To measure the effect of hypertension on neointima formation after balloon injury of rat aorta and its association with the local angiotensin converting enzyme (ACE) concentration. Balloon angioplasty of the thoracic aorta using a 2 French Fogarty catheter was performed in spontaneously hypertensive rats (SHR) and normotensive Sprague-Dawley (SD) rats. RESULTS: The injured aortic wall of SHR had already significantly higher ACE concentrations than did the uninjured aortic wall of normotensive SD rats (media: 729 +/- 37 dpm/mm2 in SHR versus 496 +/- 38 dpm/mm2 in SD rats, P < 0.01; intima: 83 +/- 5 dpm/mm2 versus 68 +/- 6 dpm/mm2 in SD rats, P < 0.01). Fourteen days after injury of the aorta the hypertensive rats had significantly higher neointima: media ratios than did the normotensive rats (0.83 +/- 0.09 versus 068 +/- 0.01, P < 0.01). This was associated with a significant increase in vascular media and neointima ACE concentrations in SHR (media 965 +/- 25 dpm/mm2, neointima 614 +/- 48 dpm/mm2) compared with those in normotensive SD rats after balloon angioplasty (media 669 +/- 23 dpm/mm2, neointima 287 +/- 33 dpm/mm2, P < 0.01). ACE inhibitor treatment with 10 mg/kg body weight lisinopril daily for 14 days by gavage reduced neointima proliferation in hypertensive and normotensive rats (neointima: media ratio: 0.35 +/- 0.02 for SHR, P < 0.01, versus untreated SHR with balloon injury; 0.28 +/- 0.01 for SD, P < 0.01, versus untreated SD rats with balloon injury). This was associated with significant vascular media ACE inhibition (SHR 149 +/- 9 dpm/mm2; SD rats 118 +/- 7 dpm/mm2; P < 0.01 versus untreated controls with balloon injury) and neointima ACE inhibition (SHR 73 +/- 4 dpm/mm2, SD rats 63 +/ 7 dpm/mm2, P < 0.01, versus untreated controls with balloon injury), but also lowered the blood pressure in SHR significantly (to 148 +/- 5 mmHg, P < 0.01, versus untreated SHR with balloon injury). When this drop in blood pressure was prevented by feeding the rats a high-salt diet (SHR with ACE inhibitor plus high salt-diet group blood pressure 193 +/- 3 mmHg, P = 0.57, versus untreated SHR) hypertension per se without the local ACE increase (ACE concentration in SHR with ACE inhibitor high-salt diet rats' media 167 +/- 10 dpm/mm2 and neointima 81 +/- 9 dpm/mm2) had only a mild effect on neointima formation after balloon angioplasty (neointima: media ratio 0.4 +/- 0.01 for SHR with ACE inhibitor plus high-salt diet versus 0.35 +/- 0.02 for SHR with ACE inhibitor plus normal-salt diet P < 0.05). Treatment with 10 mg/kg body weight angiotensin II subtype 1 receptor antagonist losartan potassium daily for 14 days by gavage was associated with a reduction in neointima formation similar to that observed with the ACE inhibitor both for SHR and for SD rats (neointima: media ratio 0.32 +/- 0.04 for SHR with losartan, 0.27 +/- 0.03 for SD rats with losartan; P < 0.01, versus untreated controls with balloon injury) suggesting that ACE inhibitor prevented neointima formation, at least in part by, reducing the local production of angiotensin II. CONCLUSION: Neointima formation after balloon angioplasty in SHR is increased compared with that in normotensive SD rats. This is due mainly to there being a higher degree of activation of the renin-angiotensin system in the aorta of the SHR before and after balloon injury compared with that in normotensive SD rats measured in terms of the increased vascular ACE concentrations. Blood pressure alone had only a moderate effect on neointima formation. PMID- 9218185 TI - Swimming training lowers the resting blood pressure in individuals with hypertension. AB - BACKGROUND: Despite the fact that swimming is often recommended for the prevention and treatment of hypertension, no study has examined the potential efficacy of regular swimming exercise for lowering the blood pressure in hypertensive humans. OBJECTIVE: To test the hypothesis that regular swimming exercise lowers the resting blood pressure. DESIGN: A 10-week closely supervised swimming training program compared with a non-exercising control group. PATIENTS: Eighteen previously sedentary men and women [aged 48 +/- 2 years (mean +/- SEM)] with stage 1 or 2 essential hypertension. RESULTS: The resting heart rated, an index of cardiovascular adaptation, decreased in the swimming training group from 81 +/- 4 to 71 +/- 3 beats/min (P < 0.01). The body mass and body fat percentage did not show statistically significant changes. The systolic blood pressure of patients in the seated position fell significantly (P < 0.05) from 150 +/- 5 to 144 +/- 4 mmHg. The seated diastolic blood pressure did not change significantly. A similar magnitude of reductions in systolic blood pressure (P < 0.05) was also found in patients in the supine position. No significant changes in plasma catecholamine concentrations, casual forearm vascular resistance, plasma volume and blood volume were observed. There were no significant changes in any of these variables in the control group. CONCLUSION: Swimming training elicits significant reductions in arterial blood pressure at rest in individuals with hypertension. This is a clinically important finding since swimming can be a highly useful alternative to land-based exercises for hypertensive patients with obesity, exercise-induced asthma, or orthopedic injuries. PMID- 9218186 TI - Reduction in external K causes increased action potential shortening in ventricular myocytes from the spontaneously hypertensive rat. AB - OBJECTIVES: (1) To study the effect of low external K and combined low K plus low Mg on the action potential of hypertrophied left ventricular myocytes isolated from the spontaneously hypertensive rat (SHR) and cells from normotensive control rats (NCR). (2) To identify differences in the response of SHR and NCR ventricular myocytes to low K and low K plus low Mg that could contribute to the increased number of arrhythmias observed in hypertrophied hearts. METHODS: Cells were superfused with Tyrode's solution containing 6 mmol/l K and stimulated at 1 Hz. Action potentials were recorded using the patch clamp technique. The effect of low K (2.4 mmol/l) and combined low K plus low Mg (2.4 mmol/l K and nominally zero Mg) on the characteristics of the action potential was measured in nine SHR, eight Wistar-Kyoto rat and 5 Wistar rat cells. RESULTS: With 6 mmol/l K, the action potential was prolonged significantly in SHR cells at 50, 70, 90 and 95% repolarizations. Both low K and low K plus low Mg shortened the action potential significantly only at 70% repolarization in NCR cells. In contrast, both low K and low K plus Mg led to marked action potential shortening at 70 and 90% repolarizations in SHR cells (P < 0.01) and also at 50 and 95% repolarizations (P < 0.05). Combined low K plus low Mg produced no additional effect on the action potential shape either in SHR or in NCR cells compared with that produced by low K alone. Although in SHR cells low K and combined low K plus low Mg produced a greater shortening of the action potential, which extended across a longer period of repolarization, a significant prolongation of the action potential still remained with low K at 50 and 95% repolarizations in SHR cells compared with that in NCR cells. CONCLUSIONS: Low K shortened the action potential both in NCR and in SHR cells but had a much greater effect on the action potential shape in SHR cells. Low K attenuated the difference in action potential shape between SHR and NCR by reducing the longer plateau observed in SHR cells. Combined low K plus low Mg exerted an effect on the action potential shape similar to that of low K alone in all cells. This increased shortening of the action potential produced by low K in SHR cells might be relevant to the increased incidence of arrhythmias associated with left ventricular hypertrophy, perhaps by shortening the wavelength of excitation and encouraging re-entrant arrhythmias, or by increasing the heterogeneity of repolarization within the ventricle. PMID- 9218187 TI - Myocardial and forearm blood flow reserve in mild-moderate essential hypertensive patients. AB - BACKGROUND: Structural readaptation of systemic resistance-sized arterioles in response to an elevated blood pressure reduces the forearm vasodilator reserve in patients with essential hypertension. The development of a similar process at the coronary microvascular level has frequently been hypothesized, but little information about coronary remodeling during the uncomplicated stage of hypertension has been obtained, and the relationship with concomitant changes in forearm blood flow reserve is not known. OBJECTIVE: To assess the minimal myocardial resistance and its relationship with the minimal forearm resistance in a group of male patients with mild-to-moderate uncomplicated hypertension and carefully matched controls. MATERIAL AND METHODS: The minimal myocardial resistance (Rmin(myocardia), the mean arterial pressure: hyperemic myocardial flow ratio after administration of 0.84 mg/kg dipyridamole, measured by using positron emission tomography and [3N]-ammonia), minimal forearm vascular resistance (Rmin(forearm), a hemodynamic index of arteriolar structure derived from the mean blood pressure and maximal postischemic forearm blood flow by venous plethysmography), echocardiographic cardiac mass and wall thickness were measured in 25 male patients with mild-to-moderate uncomplicated essential hypertension, most of whom had previously been treated, and in seven sex- and age matched normotensive controls. RESULTS: Rmin(myocardial) (and hyperemia: baseline myocardial flow ratios) did not differ significantly between the two groups, whereas Rmin(forearm) was significantly higher in hypertensives. There was no significant intra-individual correlation between the two parameters. The left ventricular mass index was greater in patients and was related previously to Rmin(forearm) but not to Rmin(myocardial) for the overall sample. In a subgroup analysis, Rmin(forearm) values were 2SD above control values in nine patients and within the normal range in the remaining 16. The myocardial reserve was very similar in the two subgroups. CONCLUSIONS: The myocardial vasodilator reserve appeared to be preserved in these mild-to-moderate uncomplicated hypertensive patients, whereas the forearm vasodilator capacity was reduced, suggesting that the hypertensive readaptation process was not distributed homogeneously over the two vascular beds. PMID- 9218188 TI - Purification and characterization of a kinin- and angiotensin II-forming enzyme in the dog heart. AB - OBJECTIVE: To purify and characterize a kinin-forming enzyme in the dog heart and to examine the ability of this enzyme to generate angiotensin (Ang) II from Ang I. METHODS: The enzyme was isolated from heart homogenate using a diethylaminoethyl-Sepharose column, an aprotinin affinity column and a wheat germ lectin-Sepharose 6MB column. Kininogenase activity was assessed with a kinin radioimmunoassay after samples had been incubated with bovine low-molecular-mass kininogen at 37 degrees C for 1 h. Ang I-converting activity was assessed by the quantitation of Ang II formed by incubation of the sample with Ang I at 37 degrees C for 3 h, using high performance liquid chromatography. The enzyme was subjected to 12.5% sodium dodecyl sulphate-polyacrylamide gel electrophoresis, stained by Coomassie brilliant blue and transferred electrically to a membrane with glycoprotein staining. RESULTS: The purified enzyme is a glycoprotein with an apparent relative molecular mass of 65 kDa by sodium dodecyl sulphate polyacrylamide gel electrophoresis. Its kininogenase activity was approximately 20 micrograms bradykinin/h per mg protein at an optimal pH of 8.0. The enzyme also converted Ang I to Ang II at an optimal pH of 6.5. Its specific activity was approximately 2 micrograms Ang II/h per mg protein. Both activities were inhibited by aprotinin, a tissue kallikrein inhibitor. Western blot analysis using polyclonal antibody against this enzyme demonstrated that this enzyme exists both in the myocardium and in the coronary artery. CONCLUSIONS: The present study showed that the kinin-forming enzyme in the dog heart is a kallikrein-like enzyme that is different from cathepsin D, cathepsin G and chymase. It is also able to Ang I to Ang II. This enzyme might play a role in regulating myocardial perfusion, mainly by generating kinins and in part by forming Ang II. PMID- 9218189 TI - A critique of the 1996 World Health Organisation expert committee report on hypertension. PMID- 9218190 TI - Modern combination therapy: the way forward in the management of hypertensive patients. Proceedings of an international symposium. Barcelona, Spain, 1-2 November 1996. PMID- 9218191 TI - Coronary disease in hypertension: a new mosaic. AB - HYPERTENSION-ASSOCIATED ABNORMALITIES THAT PROMOTE CORONARY DISEASE: Although antihypertensive treatment has been effective in reducing premature cardiovascular mortality, the effect on various organ-specific morbid events has been unequal; the effect is much more impressive on stroke reduction than on reduction of coronary events. A student of pathophysiology would have anticipated such an outcome since blood pressure elevation is only one of multiple abnormalities in hypertension. Even in its mildest form hypertension is associated with the metabolic syndrome of dyslipidemia/insulin resistance which is conducive to early atherosclerosis. A large proportion of patients also have increased sympathetic and decreased parasympathetic tone, a constellation conducive to arrhythmias and, ultimately, to sudden death. An elevated hematocrit is also found in a substantial proportion of male patients and excessive platelet aggregability has also been described in hypertension. These hematologic abnormalities are conducive to coronary thrombosis. Angiotensin II and norepinephrine, two of the most potent trophic hormones, are frequently elevated in hypertension. The effect of these hormones on the cardiac and vascular structure further increases the predilection for negative outcomes. Left ventricular hypertrophy is a potent risk factor of coronary mortality, congestive heart failure and sudden death. Vascular hypertrophy reduces the coronary reserve and at the level of skeletal muscles contributes to the evolution of the metabolic syndrome. ORGAN-SPECIFIC HYPERTENSION TREATMENT: Because of these abnormalities we are entering a new era of treatment in hypertension. Whereas an effective fall in blood pressure remains the main goal of treatment, differential effects of various antihypertensive agents on organ-specific morbidity are being actively explored. If this research proves that certain drugs have a specific advantage in defined subgroups of patients, clinical practice will change. It is reasonable to expect that in the next century we will witness a further improvement in the impact of antihypertensive treatment on public health. PMID- 9218192 TI - Success in the treatment of hypertension: a status report. AB - BACKGROUND: Pharmacological intervention studies aimed at lowering elevated blood pressure have shown that stroke, and to a lesser degree coronary heart disease, morbidity and mortality are positively affected. These beneficial effects appear to have been obtained mainly with diuretics and beta-adrenoceptor blocking agents, as shown in a meta-analysis of 13 large prospective intervention trials in hypertension. PREVIOUS STUDIES: Benefits of pharmacological antihypertensive treatment were observed in the treatment of patients with malignant hypertension in the late 1950s and early 1960s. Many of these studies were conducted before diuretics became clinically available and long before the first reports of the antihypertensive effect of beta-blockers were published. PROSPECTIVE STUDIES: Prospective intervention trials with newer antihypertensive agents such as calcium antagonists and angiotensin converting enzyme inhibitors are currently in progress. Three recently, open, retrospective case-control studies dealing with comparisons of various antihypertensive drug classes with regard to their effect on cardiovascular mortality have attracted interest. FUTURE BENEFITS: Another question of considerable clinical importance is how far blood pressure should be lowered in order to extract the maximum benefit of treatment. This issue is currently being investigated. PMID- 9218193 TI - Low-dose combinations versus monotherapies in the treatment of hypertension. AB - RATIONALE FOR DRUG COMBINATIONS: Essential hypertension is a heterogeneous disease. Different factors might interact, in an individual patient, to increase blood pressure. This explains why a drug that lowers blood pressure by a given mechanism may normalize blood pressure in some patients but not in others. combining two drugs with complementary mechanisms of action can increase the antihypertensive efficacy. DOSAGES USED IN DRUG COMBINATIONS: When administered in combination, lower doses of the separate components are generally necessary than when the drugs are prescribed as monotherapies, which may improve tolerability. Fixed-dose combination therapy is therefore an attractive approach to the treatment of hypertensive patients. It may be an appropriate choice, not only for second-line, but also for first-line therapy. PMID- 9218194 TI - Angiotensin converting enzyme inhibitors, calcium channel blockers, and their combination in the treatment of glomerular disease. AB - BACKGROUND: Glomerular diseases may progress to end-stage renal failure via the development of glomerulosclerosis. Systemic hypertension and intraglomerular hypertension are important, although not the only, determinants of this process. Proteinuria (albuminuria) is a surrogate marker for glomerular damage and renal prognosis. EFFICACY OF ANGIOTENSIN CONVERTING ENZYME (ACE) INHIBITORS: In animal experiments and in controlled clinical studies, ACE inhibitors have proved to be effective in reducing proteinuria and in preventing glomerulosclerosis or progression to end-stage renal failure, possibly more than can be explained by their effects on blood pressure. EFFICACY OF CALCIUM CHANNEL BLOCKERS: In contrast to the uniform efficacy of ACE inhibitors, the effect of calcium channel blockers is less uniform. It depends on the model of renal damage used, the extent of the fall in systemic blood pressure and the type of calcium channel blocker used. Nevertheless, clinical studies have shown a reduction in proteinuria and at least an attenuation of progression with the use of long acting calcium channel blockers. RATIONALE FOR THE COMBINATION OF ACE INHIBITORS AND CALCIUM CHANNEL BLOCKERS: If angiotensin II influences the progression of renal failure, as is universally accepted, then the combination of an ACE inhibitor (reducing the generation of angiotensin II) and a calcium channel blocker (reducing target-organ responsiveness to angiotensin II) appears a promising one. EVIDENCE FOR THE EFFECTIVENESS OF THIS COMBINATION: In animal experiments, co-administration of an ACE inhibitor and a calcium channel blocker caused a more marked reduction in glomerulosclerosis, and this was seen in the stroke-prone spontaneously hypertensive rat model even at non-antihypertensive doses. In human diabetic nephropathy at least, proteinuria (measured as a surrogate marker of the illness) was lowered more effectively by the combination of an ACE inhibitor and a calcium channel blocker than either drug used as monotherapy despite a similar fall in blood pressure. PMID- 9218196 TI - Complementary actions and risk reduction: the rationale for combination of an angiotensin converting enzyme inhibitor with a non-dihydropyridine calcium antagonist. AB - REQUIREMENTS FOR DRUG REGISTRATION: Current drug registration procedures by the United States Food and Drug Administration (FDA) require at least two placebo controlled factorial- and/or parallel-study designs. The statistical and graphical analysis of a factorial-design study allows the investigator to identify the optimal dose of the combination but these comparisons require considerable overall patient numbers for statistical power. In contrast, parallel design studies require about 150 matched patients in each of the four arms. DRUG COMBINATIONS: In an effect to compare combination studies across antihypertensive drug classes, we examined data from studies that formed the basic of past registration applications to the FDA. Our review findings indicated that combined antihypertensive effects were, at best, approximately additive for a number of different combinations. However, this takes no account of the advantages of drug combinations in allowing reduced dosages of each drug with fewer adverse effects than monotherapy. CALCIUM ANTAGONISTS: In the case of calcium antagonists, physicians must differentiate between rapid-onset or short-acting dihydropyridines, which raise the heart rate and plasma noradrenaline, and longer acting newer calcium antagonists. Recent evidence indicates that the combination of a long-acting calcium antagonist and an angiotensin converting enzyme inhibitor is particularly effective in reducing cardiovascular risk. CONCLUSIONS: Combining drugs with different mechanisms of action seems to be a reasonable and effective way of treating hypertension. It is essential, however, to establish the efficacy and safety of all new drug combinations by performing carefully designed clinical studies that fulfill the stringent FDA requirements. PMID- 9218195 TI - Preclinical considerations and results with the combination of verapamil and trandolapril: blood pressure reduction and beyond. AB - OBJECTIVE: Preclinical studies were designed to investigate whether the combination of verapamil and trandolapril was more potent than either drug alone for the treatment of hypertension and concomitant cardiovascular or metabolic diseases. HYPERTENSION: In spontaneously hypertensive rats (SHR) oral treatment with the combination of verapamil plus trandolapril not only significantly reduced elevated blood pressure for 24 h but, after terminating treatment, the antihypertensive effect lasted longer than 48 h, indicating potentiation of each compound's effect. In stroke-prone SHR, life-long oral treatment with low doses of verapamil, trandolapril or their combination significantly prolonged life in each case, but results obtained with the combination were additive compared with monotherapy. HYPERTENSION AND DIABETES: In an animal model of non-insulin dependent diabetes, the obese Zucker rat, 2 weeks oral treatment with verapamil, trandolapril or both combined significantly improved leukocyte rheology, but only the combination treatment normalized the microcirculation. In insulin-dependent diabetes (streptozotocin-induced) in SHR, oral treatment with the combination for 12 weeks additively reduced blood pressure and totally normalized proteinuria and albuminuria. Verapamil was least effective. Trandolapril showed intermediate protection against insulin-dependent diabetes-related renal failure. CHRONIC RENAL FAILURE: In SHR, almost in parallel with the progression of hypertension, progressive renal failure develops, as indicated by increasing proteinuria and albuminuria. With lifelong treatment in these rats, verapamil + trandolapril combined inhibited the progression towards end-stage renal failure more effectively than either compound used as monotherapy and also significantly prolonged survival. In a second study, in older rats with hypertension and proteinuria, chronic treatment with the combination reduced renal insufficiency in parallel with a reduction in glomerulosclerosis independently of a blood pressure reduction and significantly more potently than each drug alone. CORONARY ARTERY DISEASE: In pigs subjected to consecutive left anterior descending artery occlusions, intravenous verapamil dose-dependently reduced ischaemic myocardial K+ loss. Trandolapril alone was ineffective. Combining two ineffective doses (0.02 mg/kg verapamil plus 0.1 mg/kg trandolapril) led to a significant, additive reduction in extracellular K+ concentrations during ischaemia. Combined intracoronary application of 0.5 mg/kg trandolapril and 0.1 mg/kg verapamil before occlusion in dogs improved myocardial contractility 3 h after reperfusion to a greater extent than monotherapy and reduced the incidence of reperfusion arrhythmias. In dogs exposed to hypoxaemia after ischaemic reperfusion injury, paradoxical coronary constriction was observed. Treatment with trandolapril (0.05 mg/kg) plus verapamil (0.1 mg/kg) inhibited this vasoconstriction in response to repeated hypoxia. In anaesthetized dogs, cyclic coronary flow reductions were induced. Trandolapril reduced these by 70-80% from a dose of 0.1 mg/kg onwards. Verapamil (0.2 mg/kg) had no effect. Combining ineffective doses of trandolapril (0.05 mg/kg) and verapamil (0.1 mg/kg) led to a 70% fall in the flow reductions. Further experiments using either the bradykinin B2 antagonist HOE 140 or the angiotensin II AT1-receptor antagonist Exp 3174 showed that this effect was not mediated by increases in bradykinin levels but by a decrease in angiotensin II. CONGESTIVE HEART FAILURE: Two weeks after occlusion of the left anterior descending artery, surviving rats were randomly allocated to no treatment or to verapamil at 20 or 40 mg/kg per day or to trandolapril at 0.3 or 0.7 mg/kg per day or to combinations of both drugs, orally for 24 weeks. (ABSTRACT TRUNCATED) PMID- 9218197 TI - Fixed low-dose combination of an angiotensin converting enzyme inhibitor and a calcium channel blocker drug in the treatment of essential hypertension. AB - AIM: To review the possibilities of combination treatment in hypertension with special emphasis on the combination of an angiotensin converting enzyme (ACE) inhibitor and a calcium channel blocker. RESULTS: An increasing number of combination of drugs has become available. Fixed combinations, often low-dose, have proved suitable for the treatment of hypertension, even as first-line therapy. CONCLUSION: A fixed combination of an ACE inhibitor and a calcium channel blocker, in particular the combination of trandolapril and verapamil, is a suitable alternative to existing treatment modalities. PMID- 9218198 TI - Assessment of antihypertensive treatment by ambulatory blood pressure. AB - ADVANTAGES OF AMBULATORY BLOOD PRESSURE MONITORING: Ambulatory blood pressure monitoring is now used widely to assess the efficacy of antihypertensive drugs in daily life conditions. These 24-h measurements have a number of advantages compared to conventional sphygmomanometric readings. Although a small placebo effect is observed in the first few hours after placebo administration, 24-h average blood pressure is substantially devoid of any placebo effect. Moreover, ambulatory blood pressure is not affected by the alerting reaction usually observed during the doctor's visit. When the 24-h average value is considered, ambulatory blood pressure is more reproducible than clinic blood pressure. Finally, ambulatory blood pressure is prognostically more important than clinic blood pressure, since the end-organ damage associated with hypertension is more closely related to 24-h than to clinic blood pressure. Ambulatory blood pressure monitoring is therefore particularly useful when testing the efficacy of new antihypertensive agents on 24-h blood pressure. TESTING THE COMBINATION OF VERAPAMIL AND TRANDOLAPRIL: In a recent study we evaluated the efficacy of a fixed combination of verapamil and trandolapril using both clinic and ambulatory blood pressure measurements. Ambulatory blood pressure monitoring showed that the effect of the combination of verapamil and trandolapril was greater than the effect of either of the two drugs administered alone. However, the clinic blood pressure measurements failed to show any systemically greater effect with the combination versus monotherapy. This further indicates that ambulatory blood pressure is superior to conventional blood pressure in the assessment of antihypertensive drugs. PMID- 9218199 TI - Safety profile of the combination of verapamil and trandolapril. AB - ADVANTAGES OF DRUG COMBINATIONS: The particular advantage usually sought with antihypertensive drug combinations is an improvement in blood pressure control. However, at least as important is a consideration of adverse reactions and the safety of the combination compared to monotherapy. VERAPAMIL AND TRANDOLAPRIL: The fixed-dose combination of the calcium antagonist verapamil and the angiotensin converting enzyme (ACE) inhibitor trandolapril consists of a sustained-release (SR) tablet formulation of verapamil, and an instant-release granulation of trandolapril, a prodrug of the active metabolite trandolaprilat. The two drugs are suitable for combined administration since the half-life, time to maximal plasma concentration and peak: trough ratio are very similar for each drug. ADVERSE EFFECT PROFILE: In randomized placebo-controlled studies adverse events were observed in 25.6% of patients on placebo, 34.2% on verapamil SR, 27.3% on trandolapril and 27.9% on the combination of 180 mg verapamil + 2 mg trandolapril per day. Constipation and cough, which are considered to be specific adverse effects, occurred in 3.4% and 2.4% of patients on monotherapy with verapamil and trandolapril, respectively, but in only 2.9% and 1.9% of patients on combination therapy, respectively. Electrocardiograph recordings have not shown a prolongation of the P-Q interval to > 200 ms (placebo 1.1%, verapamil 1.2%, trandolapril 1,8%, combination 1.2%). No specific changes were found in the laboratory parameters. CONCLUSIONS: In summary, the profile of adverse drug reactions of a fixed-dose combination of verapamil and trandolapril consists the typical side effects of the monocompounds. The frequencies of adverse events were equal to or even lower than those for the monocompounds or of placebo. PMID- 9218200 TI - Diabetes and hypertension: the bad companions. AB - DIABETES AND HYPERTENSION: Diabetes mellitus and hypertension are interrelated diseases that strongly predispose people to atherosclerotic cardiovascular disease. Hypertension is about twice as frequent in individuals with diabetes as in those without. The prevalence of coexisting hypertension and diabetes appears to be increasing in industrialized nations because populations are aging, and both hypertension and non-insulin-dependent diabetes mellitus (NIDDM) increase with age. An estimated 35-75% of diabetic cardiovascular and renal complications can be attributed to hypertension. ESSENTIAL HYPERTENSION: Essential hypertension accounts for the majority of hypertension in individuals with diabetes, particularly those with NIDDM, who constitute over 90% of those with a dual diagnosis of diabetes and hypertension. Diabetic nephropathy, which occurs after 15 years of diabetes in one-third of those with insulin-dependent diabetes and 20% of those with NIDDM, is an important contributing factor to the development of hypertension in the diabetic. New investigations should focus increasingly on identifying appropriate antihypertensive agents that not only lower blood pressure but also reduce cardiovascular risk and retard the rate of progression of diabetic renal disease. PMID- 9218201 TI - How far should blood pressure be reduced in diabetic hypertensive patients? AB - EPIDEMIOLOGY OF DIABETES: Diabetes mellitus and arterial hypertension are closely related diseases that strongly predispose an individual to atherosclerotic cardiovascular disease and to renal failure. High blood pressure is twice as frequent in diabetics compared with the general population, and often precedes and contributes to the development of diabetic nephropathy. The prevalence of coexisting arterial hypertension and non-insulin-dependent diabetes mellitus (NIDDM) is increasing as populations age, giving an increased prevalence of both diseases. TREATMENT OF HYPERTENSIVE DIABETIC PATIENTS: The goal of treating arterial hypertension in diabetic patients is to prevent death and disability associated with high blood pressure. In addition, other reversible risk factors for cardiovascular disease, seen so frequently in hypertensive diabetics, also need to be addressed. The optimal goal of blood pressure control in diabetics has not been established, but there are indications that it should be lower than the 130/85 mmHg systolic/diastolic pressure recommended by current guidelines. In the presence of multiple associated risk factors, most guidelines suggest a threshold for intervention of > or = 140/90 mmHg. In particular, in hypertensive diabetic patients intervention must be early and aggressive. PMID- 9218202 TI - Requirements for antihypertensive therapy in diabetic patients: metabolic aspects. AB - AIM OF ANTIHYPERTENSIVE TREATMENT IN DIABETICS: Prevention or treatment of hypertensive in diabetic patients reduces the incidence and progression of diabetic complications of retinopathy and nephropathy, cerebro- and cardio vascular disease, and widespread macroangiopathy. Therefore, in patients with diabetes and hypertension beside good glucose control, the basic and probably major intervention steps is to normalize blood pressure. Antihypertensive treatment usually means life-long use of antihypertensive drugs. METABOLIC EFFECTS OF DIFFERENT DRUG CLASSES: Given the known diabetogenic properties of several antihypertensive drugs and their high rate of use, in probably a substantial proportion of patients with diabetes or prone to develop diabetes, treating arterial hypertension with conventional diuretics and/or beta-blockers might, in the long term, offset the beneficial effects of lowering blood pressure. Furthermore, there are conflicting reports of increased mortality in patients treated with diuretics, beta-blockers or calcium antagonists. Consequently, metabolic aspects and side effects of antihypertensive drugs are key elements in determining the preference for a specific antihypertensive regimen. Although the impact of hyperinsulinemia/insulin resistance on morbidity and mortality is an open question, it is preferable that antihypertensive treatment does not increase insulin resistance and/or hyperinsulinemia. Chronic beta-blocker treatment can be accompanied by an increase in insulin resistance. Calcium antagonists and angiotensin converting enzyme (ACE) inhibitors and alpha(1)-blockers are neutral or might even improve insulin resistance and lipid profile. Thiazides impair glucose tolerance, increase low-density lipoprotein cholesterol and decrease potassium, although these side effects are dose dependent. Unless diuretics are needed for reasons other than hypertension, treatment of diabetics with thiazides should be avoided until the influence of these agents on prognosis is clarified. If the addition of a diuretic is needed, the metabolically neutral indapamide would seem a reasonable choice. PREFERRED FIRST-LINE TREATMENT: On the basis of favorable pharmacological profiles, ACE inhibitors and certain calcium antagonists have emerged as the preferred first line drugs in the treatment of the hypertensive diabetic patient. In diabetics with nephropathy, therapy is usually initiated with an ACE inhibitor. Moreover, the combination of an ACE inhibitor and a calcium antagonist that lowers the heart rate (such as verapamil) might offer even greater advantages than either class of drug alone, since they combine metabolic neutrality with added antihypertensive and renal protective efficacy. PMID- 9218203 TI - Socioeconomic impact of diabetic nephropathy: can we improve the outcome? AB - BACKGROUND: The prevalence of non-insulin-dependent diabetes mellitus-associated nephropathy is increasing worldwide. Obviously, a greater commitment of time is required from health providers to care for such patients. Moreover, when these patients develop end-stage renal disease, healthcare costs increase geometrically when viewed in the total context of lost wages and increased health-care expenditures. INTERVENTIONS: Strict control of glucose as well as a low-protein and low-salt diet are important, but ultimately, aggressive blood pressure reduction is required to markedly decrease the time to dialysis. TREATMENT: Angiotensin converting enzyme (ACE) inhibitors should be part of the blood pressure-lowering therapy in all such patients. Recent data support the concept that addition of an ACE inhibitor to other blood pressure-lowering regimens delays the time to dialysis. Moreover, non-dihydropyridine calcium channel blockers should also be added for blood pressure control in these patients. STUDIES: Recent evidence from long-term studies in patients with nephropathy from non-insulin-dependent diabetes suggest that this subclass of calcium blockers is similar in efficacy to ACE inhibitors. CONCLUSIONS: The use of these strategies to reduce arterial systolic/diastolic pressure to < 130/80 mmHg will provide long term benefit both to the patient and to society. PMID- 9218204 TI - Can we further slow down the progression to end-stage renal disease in diabetic hypertensive patients? AB - HYPERTENSION AND DIABETES: Hypertension occurs about twice as frequently in diabetics compared with the general population, with a prevalence of approximately 25% in young patients with insulin-dependent diabetes mellitus (IDDM) and 50% in newly diagnosed non-IDDM (NIDDM) patients. Studies strongly suggest that hypertensions is involved in the progression and perhaps the onset of diabetic nephropathy, which is a major cause of illness and premature death in diabetic patients, largely through accompanying cardiovascular disease and end stage renal failure. TREATMENT: A large body of evidence has accumulated which emphasizes the beneficial effects of antihypertensive treatment in reducing proteinuria and preserving renal function in both IDDM and NIDDM. It appears that angiotensin converting enzyme inhibitors and certain calcium antagonists, notably the non-dihydropyridine type and second-generation dihydropyridine calcium antagonists, produce a more beneficial effect on nephropathy in terms of reducing proteinuria and slowing progression in renal failure. These drugs have displayed a nephroprotective capacity beyond their systemic blood pressure-lowering effects, and initial clinical trials with combinations of different antihypertensive drug classes have revealed additive effects in reducing albuminuria together with the lowest rate of decline in glomerular filtration rates with the lowest incidence of adverse effects. AVAILABLE STUDIES: The studies available on antihypertensive treatment in IDDM and NIDDM patients with incipient or overt diabetic nephropathy are mainly prospective. There have also been some preliminary trials with antihypertensive combinations in diabetic patients. PMID- 9218205 TI - Large artery stiffness in hypertension. AB - EFFECTS OF HYPERTENSION ON LARGE ARTERIES: The mechanical properties of large arteries make a major contribution to cardiovascular haemodynamics through the buffering of stroke volume and by propagation of the pressure pulse. A sustained increase in blood pressure often leads to stiffness of the large arteries, especially when other risk factors are present. The increased stiffness, in turn, aggravates hypertension by increasing systolic blood pressure and can induce cardiac hypertrophy and arterial lesions. Epidemiological studies strongly suggest that subjects with stiffer arteries have a high pulse pressure, and that stiffening of large arteries is associated with excess morbidity and mortality independently of other cardiovascular risk factors. ENVIRONMENTAL AND GENETIC FACTORS: Apart from high blood pressure and ageing, various environmental and genetic factors that influence the composition of the extracellular matrix of the arterial wall can increase arterial stiffness. Clinical studies suggest that the presence of some genotypes may be a particularly important risk marker for arterial stiffness, and may modulate the effects of hypertension, ageing and lipids on large arteries. EFFECTS OF ANTIHYPERTENSIVE DRUGS: The development of accurate, non-invasive methods has now made it possible to detect alterations of the large arteries. Among antihypertensive drugs, angiotensin converting enzyme inhibitors and calcium channel blockers have proved to be highly effective in improving large artery compliance, and have shown no adverse effects on metabolic factors that can alter arterial structure and function such as lipids, plasma glucose and insulin tolerance. Therefore these drugs may be particularly suitable for treating patients with increased arterial stiffness. Finally, a determination of genotypes may be helpful in the future in choosing antihypertensive therapy. PMID- 9218206 TI - Non-invasive evaluation of arterial abnormalities in hypertensive patients. AB - ARTERIAL ABNORMALITIES IN HYPERTENSION: Morbidity and mortality in hypertension are mainly determined by arterial lesions which may occur in different regional circulations (e.g. kidney, cerebral, coronary circulations, causing nephro angiosclerosis, stroke or myocardial infarction, respectively). Despite arterial heterogeneity, structural and functional abnormalities are usually observed at an early stage of hypertension in both large and small arteries. These alterations modify physiological and mechanical properties of the arterial wall, which may become clinically evident by increasing arterial pulsatility or pulse pressure; the alterations facilitate the establishment and progression of atherosclerosis and arteriosclerosis. METHODS OF ASSESSING ARTERIAL ABNORMALITIES: Several non invasive techniques can be used to assess haemodynamic properties of arteries: (1) casual and ambulatory blood pressure measurements can be used to evaluate pulse pressure; (2) pulse pressure can be measured directly in different sites of the arterial tree using the Tonometer device; (3) ultrasound techniques can be applied, including Doppler signals to assess the arterial flow, video-echo signals to analyse the arterial structure such as the intimal-medial thickness and echo-tracking systems for direct measurements of arterial wall distension and thickness; (4) pulse wave velocity is widely used as index of arterial distensibility; this parameter, assessed by the Complior device, has shown that hypertensive patients have decreased arterial distensibility and that antihypertensive treatment does not always reverse this abnormality. TREATMENT: It is important to evaluate the effect of cardiovascular risk-reduction measures on the arterial wall. Large therapeutic trials are necessary to show whether an evaluation of arterial abnormalities can identify patients with a high cardiovascular risk and contribute to their treatment and prognostic improvement. PMID- 9218207 TI - Angiotensin converting enzyme inhibitor-calcium antagonist combination: an alliance for cardioprotection? AB - MECHANISMS OF ACTION IN SMOOTH MUSCLE: Calcium antagonists and angiotensin converting enzyme (ACE) inhibitors act synergistically in reducing blood pressure. Calcium antagonists counter an excess of calcium entry through the voltage-operated channels of the vascular smooth muscle. ACE inhibitors reduce the vasoconstrictive properties of local and circulating angiotensin II. In addition, they improve endothelium-dependent vasodilation by increasing the expression of endothelial inositol by a bradykinin-related mechanism. Thus, at the smooth muscle level, calcium antagonists cause dilation by reducing external calcium entry and ACE inhibitors cause dilation by reducing internal calcium cycling and improving nitric oxide production. EFFECTS AT MOLECULAR LEVEL: The two agents have synergistic antagonists exert a potent cardioprotective action; this effect requires prophylactic administration and relies on the ATP-sparing capacity of these drugs. Moreover, prophylactic administration of verapamil but not to other calcium antagonists in patients who have suffered an acute myocardial infarction reduces overall mortality, provided no overt heart failure is present. The molecular effect of ACE inhibitors on the ischaemic heart is less well known but seems to be related to a reduction in noradrenaline release or to an improvement in bradykinin; the effect is thus complementary to that of calcium antagonists. OTHER ACTIONS OF ACE INHIBITORS: There is evidence that ACE inhibitors may have additional cardioprotective properties in being able to improve coronary flow, prevent arrhythmias, reduce the toxicity of oxygen free radicals, improve myocardial energy metabolism and prevent remodelling of the heart. ACE inhibitors can also protect the ischaemic heart by inhibiting bradykinin breakdown and by interfering with the central and peripheral nervous and hormone systems such as the kallikrein-kinin, prostaglandin and sympathetic nervous systems. The administration of ACE inhibitors to patients with acute myocardial infarction failed to reduce mortality. However, prophylactic administration to patients with myocardial infarction and heart failure resulted in a significant reduction in mortality and in hospitalization for cardiac disease. CONCLUSIONS: The combination of verapamil with an ACE inhibitor is promising not only for the treatment of hypertension, but also for ischaemic heart disease. PMID- 9218208 TI - Treatment with verapamil and trandolapril in patients with congestive heart failure and myocardial infarction. The Danish Verapamil Infarction Trial (DAVIT Study Group). AB - EFFECTS OF VERAPAMIL AND TRANDOLAPRIL: Progression of heart failure, sudden death and death from re-infarction are the major cause of the increased mortality in postinfarct patients with congestive heart failure. Angiotensin converting enzyme (ACE) inhibitors such as trandolapril can prevent the progression of heart failure and thus improve survival. The calcium antagonist verapamil has been shown to prevent sudden death and re-infarction in postinfarct patients without congestive heart failure. HYPOTHESIS: The Danish Verapamil Infarction Trial (DAVIT) study group hypothesized the combined treatment with trandolapril and verapamil might prevent cardiac events in postinfarct patients with coronary heart disease. The first double-blind randomized trial included 100 patients and supported this hypothesis, as the cardiac event rate was significantly lower after 3 months in patients treated with the combination than in those treated with trandolapril alone (14 versus 35%, respectively; P = 0.01, hazard ratio 0.35, 95% confidence interval 0.15-0.85). PMID- 9218209 TI - Thrombin-dependent calcium signalling in single human erythroleukaemia cells. AB - 1. A combination of single cell fluorescence and patch clamp techniques were used to study the mechanisms underlying thrombin-evoked Ca2+ signals in human erythroleukaemia (HEL) cells, a leukaemic cell line of platelet-megakaryocyte lineage. 2. Thrombin caused a transient increase in intracellular Ca2+ ([Ca2+]i), consisting of both release of Ca2+ from intracellular stores and influx of extracellular Ca2+. Mn2+ quench studies indicated that the thrombin-evoked divalent cation-permeable pathway was activated during, but not prior to, release from internal stores. 3. Thapsigargin (1 microM) irreversibly released internal Ca2+ from the same store as that released by thrombin and continuously activated a Ca(2+)-influx mechanism. The amplitude of the thrombin- and thapsigargin induced Ca2+ influx displayed a marked single cell heterogeneity which showed no correlation with the size of the store Ca2+ transient. 4. In whole-cell patch clamp recordings, both thrombin and thapsigargin evoked an inwardly rectifying Ca2+ current which developed with little or no increase in current noise, showed no reversal in the voltage range -110 to +60 mV and was blocked by 1 mM Zn2+. The apparent divalent cation permeability sequence of this pathway was Ca2+ > > Ba2+ > Mn2+, Mg2+. The thapsigargin-evoked current density at -100 mV varied between 0.42 and 2.1 pA pF-1 in different cells. Thrombin failed to activate additional Ca2+ current if it was added after the thapsigargin-induced inward current had fully developed. 5. These studies indicate that thrombin activates Ca2+ influx in HEL cells entirely via a Ca(2+)-store-release-activated Ca2+ current (Icrac) rather than via receptor-operated or second messenger-dependent Ca2+ channels. The level of expression of Icrac appears to be a major factor in determining the duration of the thrombin-evoked [Ca2+]i response and therefore represents a means by which cells can exert control over [Ca2+]i-dependent events. PMID- 9218210 TI - Regulation of the cytosolic Ca2+ concentration by Ca2+ stores in single smooth muscle cells from rat cerebral arteries. AB - 1. There is no general agreement on the presence or role of Ca(2+)-induced Ca2+ release in smooth muscle. In this paper, Ca(2+)-induced Ca2+ release has been investigated in rat resistance-sized superior cerebral arteries to determine its role in regulating the cytosolic Ca2+ concentration ([Ca2+]i). 2. Pressurized superior cerebral arteries developed spontaneous oscillations in diameter. These oscillations were abolished by ryanodine (an inhibitor of Ca(2+)-induced Ca2+ release) and removal of extracellular Ca2+. This suggests, indirectly, that Ca(2+)-induced Ca2+ release may regulate [Ca2+]i in the resistance arteries. 3. To determine if Ca(2+)-induced Ca2+ release could regulate [Ca2+]i, single smooth muscle cells were isolated from the superior cerebral artery, voltage clamped in the whole cell configuration and high temporal resolution [Ca2+]i measurements made. The relationship between the Ca2+ current (ICa) and rise in [Ca2+]i was examined. 4. Depolarization triggered ICa and increased [Ca2+]i. The time course of the measured increase in [Ca2+]i closely followed the increase in [Ca2+]i expected from the time-integrated ICa, although about 140-fold more Ca2+ entered the cytosol than appeared as free Ca2+. When the cells were dialysed with ryanodine (30 microM), the Ca2+ transient evoked by the ICa was substantially reduced indicating that Ca2+ influx triggered Ca2+ release from an internal store. 5. Voltage pulses to negative membrane potentials were more effective in triggering Ca2+ release than pulses to positive potentials suggesting that the Ca(2+)-induced Ca2+ release was voltage dependent. However, the release of Ca2+ from the internal store triggered by caffeine was voltage independent. These results suggest that the voltage dependence of Ca2+ release is indirect and possibly related to the plasmalemma unitary Ca2+ current magnitude. 6. The results establish that Ca(2+)-induced Ca2+ release contributes to depolarization evoked increases in [Ca2+]i in rat resistance-sized superior cerebral arteries over the physiological [Ca2+]i range (100-200 nM). Compared with more positive membrane potentials the efficacy of Ca2+ in triggering release is high at physiological membrane potentials. PMID- 9218211 TI - Effect of the immunosupressant FK506 on excitation-contraction coupling and outward K+ currents in rat ventricular myocytes. AB - 1. We examined the effects of the immunosupressant drug FK506 on excitation contraction coupling in isolated rat ventricular myocytes. [Ca2+]i transients were recorded using the intracellular Ca2+ indicators fluo-3 and indo-1 while action potentials (APs) or membrane currents were recorded using patch-type microelectrodes in the whole cell mode. 2. FK506 (25 microM) rapidly and reversibly increased the magnitude of the [Ca2+]i transient in intact cells without changing resting [Ca2+]i or the kinetics of the [Ca2+]i transient, a finding consistent with previous reports that investigated the actions of FK506 on the sarcoplasmic reticulum Ca2+ release channel. 3. The 36% increase in the [Ca2+]i transient produced by FK506 was accompanied by a 293% increase in AP duration (by 293%). Importantly, the addition of FK506 had no effect on the [Ca2+]i transient when the depolarizing duration was controlled in voltage clamp experiments. The increased AP duration could be explained by a marked inward shift in the net membrane current that was observed in these experiments. 4. The net inward current change was not directly responsible for a change in Ca2+ influx, since no change in L-type Ca2+ current (ICa) was observed. Instead, FK506 inhibited both the transient outward K+ current (Ito) and the delayed rectifier K+ current (IK). 5. We conclude that FK506 increases the [Ca2+]i transient during normal contractions by an indirect action: it prolongs the action potential. This action does not appear to depend on the established action of FK506 on the ryanodine receptor. Instead, the inhibition of outward K+ currents prolongs the AP which secondarily increases Ca2+ influx and/or decreases Ca2+ efflux. PMID- 9218212 TI - Stimulation of anion secretion by beta-adrenoceptors in the mouse endometrial epithelium. AB - 1. Regulation of anion secretion by adrenoceptors in primary culture of mouse endometrial epithelium was investigated using the short circuit current (ISC) technique. 2. Adrenaline stimulated a sustained increase in the ISC in a concentration-dependent manner. The adrenaline-induced ISC could be inhibited by pretreatment with diphenylamine 2,2'-dicarboxylic acid (DPC) or replacement of external Cl- and HCO3-, but not by amiloride or replacement of Na+ in apical solution. 3. The concentration-dependent responses of the adrenaline-induced ISC to the Cl- channel blockers glibenclamide and DPC were examined and exhibited IC50 values of 380 and 960 microM, respectively. 4. The effect of various adrenoceptor agonists on the ISC was examined. The order of potency appeared to be isoprenaline > adrenaline > noradrenaline, while no response was elicited by the alpha-adrenoceptor agonist methoxamine, indicating a predominant involvement of beta-adrenoceptors. 5. The beta-adrenoceptor antagonist propranolol was found to be much more effective than the alpha-adrenoceptor antagonist phentolamine in inhibiting the ISC responses induced by all adrenoceptor agonists examined. 6. The effect of adrenaline on the ISC was mimicked by an adenylate cyclase activator, forskolin, but suppressed by the adenylate cyclase inhibitor MDL 12,330A, indicating the involvement of cAMP. 7. Our results demonstrate that anion secretion by the mouse endometrial epithelium is regulated by beta adrenoceptors and involves a cAMP-dependent mechanism. PMID- 9218213 TI - Effects of acetylcholine on the Na(+)-K+ pump current in guinea-pig ventricular myocytes. AB - 1. The whole-cell patch clamp technique was used to study the effects of acetylcholine (ACh) on Na(+)-K+ pump current (Ip) in acutely isolated guinea-pig ventricular myocytes. Studies were performed in the absence and presence of the beta-agonist isoprenaline (Iso). 2. ACh had no effect on Ip at low or high [Ca2+]i at any voltage in the absence of Iso. Iso alone inhibited Ip at low [Ca2+]i and shifted the Ip-V relationship at high [Ca2+]i in a negative direction. Addition of 1 microM ACh reversed these effects of Iso. K0.5 for the effects of ACh was about 16 nM, regardless of [Ca2+]i. 3. The actions of ACh on the heart are usually mediated via muscarinic receptors. Atropine, a muscarinic antagonist, blocked the effects of ACh on Ip in the presence of Iso, suggesting that these effects are also mediated by muscarinic receptors. 4. Muscarinic receptors are usually coupled to a Gi protein, leading to inhibition of adenylyl cyclase and a reduction of cAMP levels. We have shown previously that basal levels of cAMP are very low in guinea-pig ventricular myocytes, and that a membrane-permeant cAMP analogue, chlorophenylthio-cAMP (CPTcAMP), mimics the effects of Iso. ACh did not reverse the effects of CPTcAMP, supporting the hypothesis that the effects of ACh on Ip are also mediated via inhibition of adenylyl cyclase. 5. The present results suggest that a high level of parasympathetic tone alone does not affect the activity of ventricular Na(+)-K+ pumps. However, if sympathetic tone is high, then muscarinic stimulation can reciprocally modulate Na(+)-K+ pump activity. PMID- 9218215 TI - In vivo downregulation of M2 receptors revealed by measurement of muscarinic K+ current in cultured guinea-pig atrial myocytes. AB - 1. Muscarinic K+ current (IK(ACh)) elicited by acetylcholine (ACh) was measured in guinea-pig atrial myocytes, which were either freshly isolated or cultured for up to 8 days. 2. The half-time of activation of inward IK(ACh) by a saturating concentration (10 microM) of ACh decreased from approximately 400 ms (in freshly isolated cells) to 250 ms after 6 days in culture. This was paralleled by an increase in the fast desensitizing component of IK(ACh). The density of steady state currents was not changed. Downregulation of M2 receptors by long-term treatment of isolated myocytes with carbachol in vitro had opposite effects. 3. The EC50 of ACh for the activation of steady-state IK(ACh) was reduced from 5 x 10(-7) M (day 0) to 8 x 10(-8) M (day 6). The shift in EC50 occurred with a half time of about 2 days, similar to the recovery from downregulation induced by treating atrial myocytes with carbachol in vitro. 4. The increase in sensitivity to ACh can be accounted for by an approximately 6-fold increase in the density of M2 receptors. 5. It is concluded that sensitization in culture reflects recovery from downregulation of M2 receptors due to the tonic vagal input to the heart in the intact animal. PMID- 9218214 TI - Characterization of apical potassium channels induced in rat distal colon during potassium adaptation. AB - 1. Chronic dietary K+ loading stimulates an active K+ secretory process in rat distal colon, which involves an increase in the macroscopic apical K+ conductance of surface epithelial cells. In the present study, the abundance and characteristics of K+ channels constituting this enhanced apical K+ conductance were evaluated using patch clamp recording techniques. 2. In isolated non polarized surface cells, K+ channels were seen in 9 of 90 (10%) cell-attached patches in cells from control animals, and in 247 of 437 (57%) cell-attached patches in cells from K(+)-loaded animals, with a significant (P < 0.001) shift in distribution density. Similarly, recordings from cell-attached patches of the apical membrane of surface cells surrounding the openings of distal colonic crypts revealed identical K+ channels in 1 of 11 (9%) patches in control animals, and in 9 of 13 (69%) patches in K(+)-loaded animals. 3. In isolated surface cells and surface cells in situ, K+ channels had mean slope conductances of 209 +/- 6 and 233 +/- 14 pS, respectively, when inside-out patches were bathed symmetrically in K2SO4 solution. The channels were sensitive to 'cytosolic' Ca2+ concentration, were voltage sensitive at 'cytosolic' Ca2+ concentrations encountered in colonic epithelial cells, and were inhibited by 1 mM quinidine, 20 mM TEA or 5 mM Ba2+ ions. 4. The data show that dietary K+ loading increases the abundance of Ca(2+)- and voltage-sensitive large-conductance K+ channels in the apical membrane of surface cells in rat distal colon. These channels constitute the enhanced macroscopic apical K+ conductance previously identified in these cells, and are likely to play a critical role in the active K+ secretory process that typifies this model of colonic K+ adaptation. PMID- 9218216 TI - A swelling-activated chloride current in rat sympathetic neurones. AB - 1. We have tested whether neurones show a swelling-induced Cl- current following hypotonic shock, by recording membrane current responses and cell volume changes in voltage clamped isolated rat sympathetic neurones during application of hypotonic solutions. 2. Using both whole-cell and perforated patch recording methods, hypotonic solution caused cell swelling and the activation of an inward Cl- current at -60 mV. This current showed weak outward rectification with no obvious time dependence. It was inhibited by SITS (0.3-1 mM), NPPB (30-300 microM) and niflumic acid (50-200 microM), but not by tamoxifen (10 microM). 3. Hypotonic solution did not cause a rise in intracellular Ca2+ concentration as measured by simultaneous indo-1 fluorescence. Also, neither the volume change nor Cl- current were affected by the removal of external Ca2+ or internal Ca2+ buffering to < or = 1 nM with EGTA. 4. The Cl- current was unaffected by an inhibitor of protein kinase C (PKC; GF109203X, 3 microM) or by omission of ATP from the pipette solution. 5. Cells exhibited a regulatory volume decrease during sustained exposure to hypotonic solution. This was completely inhibited by 0.5 mM niflumic acid. 6. It is concluded that osmotic swelling induces an outwardly rectifying, Ca2(+)- and PKC-independent Cl- current in these nerve cells. It is suggested that this current may be involved in volume regulatory mechanisms. PMID- 9218217 TI - Activation of f-channels by cAMP analogues in macropatches from rabbit sino atrial node myocytes. AB - 1. The action of the two diastereometric phosphorothioate derivatives of cAMP, Rp cAMPs and Sp-cAMPs, was investigated on hyperpolarization-activated 'pacemaker' current (i(f)) recorded in inside-out macropatches from rabbit sino-atrial (SA) node myocytes. 2. When superfused on the intracellular side of f-channels at the concentration of 10 microM, both cAMP derivatives accelerated i(f) activation; their action was moderately less pronounced than that due to the same concentration of cAMP. 3. The measurement of the i(f) conductance-voltage relation by voltage ramp protocols indicated that both cAMP analogues shift the activation curve of i(f) to more positive voltages with no change in maximal (fully activated) conductance. 4. Dose-response relationships of the shift of the i(f) activation curve showed that both Rp-cAMPs and Sp-cAMPs act as agonists in the cAMP-dependent direct f-channel activation. Fitting data to the Hill equation resulted in maximal shifts of 9.6 and 9.5 mV, apparent dissociation constants of 0.82 and 5.4 microM, and Hill coefficients of 0.82 and 1.12 for Sp-cAMPs and Rp cAMPs, respectively. 5. The activating action of Rp-cAMPs, a known antagonist of cAMP in the activation of cAMP-dependent protein kinase, confirms previously established evidence that f-channel activation does not involve phosphorylation. These results also suggest that the cAMP binding site of f-channels may be structurally similar to the cyclic nucleotide binding site of olfactory receptor channels. PMID- 9218218 TI - Decay of calcium transients after electrical stimulation in rat fast- and slow twitch skeletal muscle fibres. AB - 1. Calcium transients were calculated from fura-2 fluorescence signals (corrected for kinetic delays in the Ca(2+)-fura-2 reaction) from single rat skeletal muscle fibres, either fully dissociated from the fast-twitch flexor digitorum brevis (FDB) muscle or in small bundles from the slow-twitch soleus muscle. Fibres or bundles were embedded in agarose gel to inhibit movement and stimulated by single or trains of 1-2 ms electrical pulses (100 Hz, 2-400 ms train duration). 2. The rate constant of decay of [Ca2+] determined from single-exponential fits to the final decay phase of [Ca2+] after a single action potential was considerably faster in FDB fibres than in soleus fibres. As the stimulation duration increased, the rate constant of [Ca2+] decay decreased for both the FDB and soleus fibres, but the effect was greater in FDB than in soleus fibres. 3. Using the magnitude of the decline in the rate constant of [Ca2+] decay with increasing stimulation duration as an index of relative contribution of the saturable Ca2+ binding sites on parvalbumin, subpopulations termed 'high', 'medium' and 'low', referring to estimated parvalbumin content, were determined within each group of FDB and soleus fibres. In fibres assigned to the 'high' and 'medium' groups, parvalbumin was the major contributor (50-73%) to the [Ca2+] decay rate constant after a single action potential. In fibres in the 'low' group, parvalbumin contributed only 0-28% to the rate constant of [Ca2+] decay. 4. Fluorescence recordings using mag-fura-2, a lower-affinity Ca2+ indicator expected to be in equilibrium with myoplasmic Ca2+, gave similar values for both the [Ca2+] decay rate constant after a single action potential and the decrease in this rate constant with increased stimulation duration, as found for the fura-2 [Ca2+] transients from FDB and soleus fibres. Thus, the observed differences in decay rate of Ca2+ were not introduced by kinetic correction of the fura-2 recordings, but are attributed to differences in the Ca2+ binding and transport properties of fast- and slow-twitch mammalian fibres. PMID- 9218219 TI - Dual actions of tetracaine on intramembrane charge in amphibian striated muscle. AB - 1. The effects of graded concentrations of tetracaine on the steady-state and kinetic properties of intramembrane charge were examined in intact voltage clamped amphibian muscle fibres. 2. The micromolar tetracaine concentrations that were hitherto reported to abolish Ca2+ transients in skeletal muscle failed to affect significantly the steady-state charge. Maximal reductions of such intramembrane charge required relatively high, 1-2 mM, concentrations of tetracaine. 3. The plots of maximum charge against tetracaine concentration suggested a saturable 1:1 drug binding that spared a fixed amount of tetracaine resistant (q beta) charge but inhibited a discrete fraction of susceptible (q gamma) charge with a KD between 0.1 and 0.2 mM. 4. The q beta charge thus isolated by 2 mM tetracaine was conserved through a wide range of applied test voltages and pulse durations and regardless of whether the imposed transition from the holding potential (-90 mV) to the test potential took place in one or more steps. 5. Similarly, 'on' and 'off' q beta currents that were elicited by voltage steps from fixed conditioning to varying test levels mapped onto non linear phase-plane trajectories that nevertheless depended uniquely upon voltage. In contrast, the currents that followed voltage steps made from varying prepulse levels to fixed -90 or -20 mV test potentials mapped onto identical q beta phase plane trajectories that were independent of the prepulse history. 6. The charge movements that followed strong depolarizing voltage clamp steps to test potentials in the range -50 to 0 mV approximated simple monotonic decays that could empirically be described by a single time constant. Nevertheless, a complete inhibition of a tetracaine-sensitive (q gamma) charge movement by 2 mM tetracaine that left only q beta charge, sharply altered both the magnitude and the voltage dependence of these time constants. This establishes a distinct contribution of the q gamma species to overall charge kinetics even at such test voltages. 7. Under such a criterion for the voltage dependence of charging kinetics, even the micromolar (0.05-0.2 mM) tetracaine concentrations that failed to markedly alter the steady-state charge consistently increased the charging time constants yet did not influence their voltage sensitivity. 8. These findings demonstrate the existence of separate kinetic and steady-state effects of tetracaine on intramembrane charge movements, at micromolar and millimolar anaesthetic concentrations, respectively. These parallel earlier effects of tetracaine that have been reported upon the transient and sustained components of sarcoplasmic reticular Ca2+ release. They also establish that maximally effective concentrations of tetracaine isolate a single distinct species of conserved (q beta) intramembrane charge. PMID- 9218220 TI - Sarcomere length dependence of the rate of tension redevelopment and submaximal tension in rat and rabbit skinned skeletal muscle fibres. AB - 1. We examined the hypothesis that in skeletal muscle the steep relationship between twitch tension and sarcomere length (SL) within the range 2.30 to 1.85 microns involves SL-dependent alterations in the rate of tension development. 2. In skinned preparations of both rat slow-twitch and rabbit fast-twitch skeletal muscle fibres the rate of tension redevelopment (ktr) at 15 degrees C was reduced at short SL (approximately 2.00 microns) compared with a longer SL (approximately 2.30 microns). In submaximally activated fibres, the decrease in ktr over this range of lengths was greater in fast-twitch fibres (38% reduction) than in slow twitch fibres (14% reduction). 3. Ca2+ sensitivity of tension, as assessed as the pCa (-log[Ca2+]) for half-maximal activation, or pCa50, decreased to a greater extent in rabbit fast-twitch skeletal muscle fibres than in slow-twitch fibres from both rabbit and rat when SL was reduced from approximately 2.30 to approximately 1.85 microns. The delta pCa50 over this SL range was 0.24 +/- 0.07 pCa units in fast-twitch fibres from rabbit psoas muscle. The delta pCa50 for slow-twitch fibres from rabbit and rat soleus muscle was 0.08 +/- 0.02 and 0.10 +/- 0.04 pCa units, respectively. 4. Osmotic compression of both slow-twitch and fast-twitch fibres at a SL of 2.00 microns increased ktr to values similar to those obtained at a SL of 2.30 microns in the absence of dextran. This result indicates that the slower rate of tension redevelopment at short SL is due in large part to the increase in interfilament lattice spacing associated with shorter SL. 5. Taken together, these results suggest that length dependence of twitch tension is, in part, due to length dependence of isometric cross-bridge interaction kinetics, an effect that is mediated by length-dependent changes in interfilament lattice spacing. PMID- 9218221 TI - Rhythmic motor activity and interlimb co-ordination in the developing pouch young of a wallaby (Macropus eugenii). AB - 1. The forelimb motor behaviour of developing wallaby was studied. A clock-like alternating movement was reactivated whenever the animal was removed from the pouch. 2. Forelimb stepping frequency increased during the first 3 weeks of development, while the phase relationship remained constant. Forelimb activity could be affected by altering the afferent feedback from the contralateral limb, or an increase in ambient temperature. 3. In vitro experiments were performed using an isolated brainstem-spinal cord preparation from animals up to 6 weeks postnatal. Fictive locomotor activity could be evoked by electrical stimulation or bath-applied NMDA (< 10 microM). 4. Bath-applied strychnine (10-25 microM) and bicuculline (10-50 microM) disrupted the phase relationship between motor pools, while rhythmic motor discharge remained in the absence of these inhibitory pathways. 5. The present findings indicate that the pattern generator that underlies the robust forelimb movement during the first journey to the pouch is retained for different motor functions during in-pouch development. The neural network that underlies such behaviour can be divided into two major components, a rhythm generator within each hemicord, and a pattern co-ordinating pathway which involve both glycinergic and GABAergic interneurones. PMID- 9218223 TI - Evidence for the involvement of histaminergic neurones in the regulation of the rat oxytocinergic system during pregnancy and parturition. AB - 1. Previous studies have shown that histaminergic neurones of the tuberomammillary nucleus project directly to hypothalamic magnocellular nuclei and that intracerebroventricular administration of histamine increases the synthetic activity of magnocellular oxytocin neurones. 2. Histaminergic neurones of the dorsomedial tuberomammillary nucleus that project to the magnocellular region of the paraventricular nucleus are activated during late pregnancy and lactation, as measured by an increase in mRNA for the synthetic enzyme histidine decarboxylase. 3. There is a concomitant increase in oxytocin mRNA in magnocellular neurones of the paraventricular nucleus. This increase in mRNA contributes to an accumulation of oxytocin before birth and to continued oxytocin synthesis during lactation. 4. Intracerebroventricular administration of mepyramine, a specific antagonist of the H1 histamine receptor, causes a delay in the birth of subsequent pups if given to the mother during parturition. Vehicle or the H2 receptor antagonist cimetidine has no effect. Thus, histamine acts centrally, via H1 receptors, during parturition and may have an excitatory effect on oxytocin release. 5. These results suggest that afferent histaminergic neurones show increased activity during pregnancy and may be responsible for the increase of synthesis in magnocellular oxytocin neurones at this time. If, as previously reported, these histamine neurones can reduce the electrical activity of oxytocin neurones via H2 receptors, then they may have a dual effect, increasing the synthesis of oxytocin while inhibiting its premature release. At term, any inhibitory effects of histamine are overcome to allow oxytocin secretion. PMID- 9218222 TI - Fluid transport across cultures of human tracheal glands is altered in cystic fibrosis. AB - 1. There is evidence that defective submucosal gland secretion contributes to the airway pathology of cystic fibrosis (CF). Using a capacitance probe technique, we have compared fluid transport across submucosal gland cultures from individuals with and without CF. 2. Under baseline conditions, approximately 60% of non-CF cultures secreted fluid; the rest absorbed. In secreting tissues, amiloride increased secretion, whereas in absorbing tissues it reduced or reversed absorption. 5-Nitro-2(3-phenylpropylamino)-benzoate (NPPB) a blocker of the CF transmembrane conductance regulator (CFTR), converted secretion to absorption. Thus, the direction and magnitude of baseline fluid movement depended on a balance between active absorption of Na+ and cAMP-dependent secretion of Cl-. 3. 8-(4-Chlorophenylthio)-adenosine 3',5'-cyclic monophosphate (CPT-cAMP), methacholine and luminal uridine 5'-triphosphate (UTP) all induced or increased fluid secretion across non-CF cultures. Results with NPPB and with 4,4' diisothiocyanatostilbene-2,2'-disulphonate (DIDS), a blocker of Ca(2+)-activated Cl- channels, suggested that fluid secretion induced by CPT-cAMP was mediated primarily by CFTR; UTP acted entirely via Ca(2+)-activated Cl- channels, and methacholine activated both pathways. 4. All CF cultures showed baseline fluid absorption, which was abolished by amiloride. 5. CF cultures showed a normal secretory response to UTP, a reduced response to methacholine, and no response to CPT-cAMP. 6. Thus, the absorptive processes of airway glands are retained in CF, but the cAMP-dependent secretory process is lost. This would markedly reduce the water content of gland secretions. The resulting change in viscosity would contribute to the accumulation of airway mucus which is characteristic of this disease. PMID- 9218224 TI - Differing effects of histamine and serotonin on microvascular permeability in anaesthetized rats. AB - 1. We have investigated simultaneous changes in the hydraulic permeability (Lp) and the retention of perfusate macromolecules in single mesenteric venules of anaesthetized rats during perfusion with either histamine or serotonin. 2. The venules were microperfused in situ. Retention of macromolecules was assessed from the effective oncotic pressure (omega delta pi) exerted by the perfusate across the vessel walls. Lp and omega delta pi were estimated by the red cell microperfusion technique. 3. Perfusion with histamine (at concentrations between 16 microM and 3.26 mM) and serotonin (at concentrations between 26 microM and 1.3 mM) transiently increased Lp and reduced omega delta pi. Maximal changes were seen at 6-9 min with histamine and at 3 min with serotonin. 4. Maximal increases in Lp were greater with histamine (approximately 3-fold) than with serotonin (1.5 to 2-fold). Serotonin, however, decreased omega delta pi from a baseline of 14 15 cmH2O to one of 6-7 cmH2O whereas the fall of omega delta pi with histamine was only from 14-15 cmH2O to 10-11 cmH2O. 5. The data are consistent with the hypothesis that serotonin increases permeability by inducing openings in the venular endothelium which do not retain macromolecules. If histamine also increases permeability by gap formation, these gaps are able to retain macromolecules to a significant extent. PMID- 9218225 TI - Renal tubular lithium reabsorption in potassium-depleted rats. AB - 1. In order to identify the tubular sites responsible for the reduced fractional excretion of lithium (FELi) during potassium depletion, free-flow micropuncture was performed in anaesthetized rats that had been fed a low potassium (low-K+) diet or a control diet for 5-6 days. FELi in low-K+ rats was 0.09 +/- 0.02, compared with 0.25 +/- 0.01 in control animals. 2. Fractional water reabsorption in proximal convoluted tubules was enhanced in potassium-depleted rats. However, fractional lithium reabsorption was not. Consequently, the tubular fluid-to plasma lithium concentration ratio at the late proximal convoluted tubule was raised in the low-K+ animals (1.50 +/- 0.03 vs. 1.18 +/- 0.02; P < 0.001). 3. Fractional lithium delivery to the early distal tubule in low-K+ rats (0.31 +/- 0.01) was similar to that in control animals (0.30 +/- 0.02). However, whereas in control rats there was no significant difference between early and late distal tubular deliveries of lithium, late distal fractional lithium delivery in the low K+ group was reduced markedly (to 0.10 +/- 0.01). 4. Treatment of potassium depleted rats with amiloride had no effect on lithium reabsorption in the proximal convoluted tubule or loop of Henle. However, fractional lithium delivery to the end of the distal tubule was increased slightly (to 0.15 +/- 0.02; P < 0.05) and FELi was increased substantially (to 0.22 +/- 0.01; P < 0.001). 5. It is concluded that two factors contribute to the reduced FELi seen in potassium depleted rats: lithium reabsorption in the superficial distal tubules and amiloride-sensitive lithium reabsorption in the collecting ducts. The data also suggest heterogeneity with respect to lithium handling between superficial and deep nephrons during potassium depletion. PMID- 9218226 TI - Sympathetic nerve traffic correlates with the release of nitric oxide in humans: implications for blood pressure control. AB - 1. Resting human sympathetic vasoconstrictor traffic displays large reproducible inter-individual differences which are similar in nerves to muscle, heart and kidney. In spite of this there is no correlation between levels of blood pressure and sympathetic traffic. To test the hypothesis that the pressor effect of the vasoconstrictor activity is counteracted by a circulating dilating factor we measured muscle nerve sympathetic activity (MSA) and an indicator of nitric oxide release (plasma nitrate) in healthy young males. 2. Sympathetic activity was recorded with the microneurographic technique in the peroneal nerve and a forearm venous plasma sample was obtained in twenty-one normotensive males aged 21-28 years. Plasma nitrate was analysed by gas chromatography and mass spectrometry. 3. There was a positive linear correlation between the plasma nitrate concentration and the strength of MSA both when the nerve activity was expressed as bursts per minute and bursts per 100 heart beats (r = 0.51, P = 0.02 and r = 0.46, P = 0.04, respectively). 4. The data suggest that the stronger the sympathetic activity the higher the release of the dilating substance, nitric oxide. This would be expected to counteract vasoconstrictor effects of the nerve traffic and thereby contribute to the lack of relationship between resting levels of MSA and blood pressure. We speculate that altered coupling between sympathetic traffic and nitric oxide release may cause abnormal peripheral resistance, e.g. in hypertension. PMID- 9218227 TI - The interplay of central and peripheral factors in limiting maximal O2 consumption in man after prolonged bed rest. AB - 1. The effects of bed rest on the cardiovascular and muscular parameters which affect maximal O2 consumption (VO2,max) were studied. The fractional limitation of VO2,max imposed by these parameters after bed rest was analysed. 2. The VO2,max, by standard procedure, and the maximal cardiac output (Qmax), by the pulse contour method, were measured during graded cyclo-ergometric exercise on seven subjects before and after a 42-day head-down tilt bed rest. Blood haemoglobin concentration ([Hb]) and arterialized blood gas analysis were determined at the highest work load. 3. Muscle fibre types, oxidative enzyme activities, and capillary and mitochondrial densities were measured on biopsy samples from the vastus lateralis muscle before and at the end of bed rest. The measure of muscle cross-sectional area (CSA) by NMR imaging at the level of biopsy site allowed computation of muscle oxidative capacity and capillary length. 4. The VO2,max was reduced after bed rest (-16.6%). The concomitant decreases in Qmax (-30.8%), essentially due to a change in stroke volume, and in [Hb] led to a huge decrease in O2 delivery (-39.7%). 5. Fibre type distribution was unaffected by bed rest. The decrease in fibre area corresponded to the significant reduction in muscle CSA (-17%). The volume density of mitochondria was reduced after bed rest (-16.6%), as were the oxidative enzyme activities ( 11%). The total mitochondrial volume was reduced by 28.5%. Capillary density was unchanged. Total capillary length was 22.2% lower after bed rest, due to muscle atrophy. 6. The interaction between these muscular and cardiovascular changes led to a smaller reduction in VO2,max than in cardiovascular O2 transport. Yet the latter appears to play the greatest role in limiting VO2,max after bed rest (> 70% of overall limitation), the remaining fraction being shared between peripheral O2 diffusion and utilization. PMID- 9218230 TI - "Shotgunning" as an illicit drug smoking practice. AB - There has been a rise in illicit drug smoking in the United States. "Shotgunning" drugs (or "doing a shotgun") refers to the practice of inhaling smoke and then exhaling it into another individual's mouth, a practice with the potential for the efficient transmission of respiratory pathogens. Three hundred fifty-four drug users (239 from a syringe exchange and 115 from a drug detoxification program) were interviewed about shotgunning and screened for tuberculosis (TB). Fifty-nine (17%; 95% CI 12.9%-20.9%) reported shotgunning while smoking crack cocaine (68%), marijuana (41%), or heroin (2%). In multivariate analysis, age < or = 35 years (OR 2.0, 95% CI 1.05-3.9), white race (OR 1.2, 95% CI 1.2-4.8), drinking alcohol to intoxication (OR 2.2, 95% CI 1.1-4.3), having engaged in high risk sex (OR 2.6, 95% CI 1.04-6.7), and crack use (OR 6.0, 95% CI 3.0-12) were independently associated with shotgunning. Shotgunning is a frequent drug smoking practice with the potential to transmit respiratory pathogens, underscoring the need for education of drug users about the risks of specific drug use practices, and the ongoing need for TB control among active drug users. PMID- 9218228 TI - Sprint training enhances ionic regulation during intense exercise in men. AB - 1. This study investigated the effects of 7 weeks of sprint training on changes in electrolyte concentrations and acid-base status in arterial and femoral venous blood, during and following maximal exercise for 30 s on an isokinetic cycle ergometer. 2. Six healthy males performed maximal exercise, before and after training. Blood samples were drawn simultaneously from brachial arterial and femoral venous catheters, at rest, during the final 10 s of exercise and during 10 min of recovery, and analysed for whole blood and plasma ions and acid-base variables. 3. Maximal exercise performance was enhanced after training, with a 13% increase in total work output and a 14% less decline in power output during maximal cycling. 4. The acute changes in plasma volume, ions and acid-base variables during maximal exercise were similar to previous observations. Sprint training did not influence the decline in plasma volume during or following maximal exercise. After training, maximal exercise was accompanied by lower arterial and femoral venous plasma [K+] and [Na+] across all measurement times (P < 0.05). Arterial plasma lactate concentration ([Lac-]) was greater (P < 0.05), but femoral venous plasma [Lac-] was unchanged by training. 5. Net release into, or uptake of ions from plasma passing through the exercising muscle was assessed by arteriovenous concentration differences, corrected for fluid movements. K+ release into plasma during exercise, and a small net K+ uptake from plasma 1 min post-exercise (P < 0.05), were unchanged by training. A net Na+ loss from plasma during exercise (P < 0.05) tended to be reduced after training (P < 0.06). Release of Lac- into plasma during and after exercise (P < 0.05) was unchanged by training. 6. Arterial and venous plasma strong ion difference ([SID]; [SID] = [Na+] + [K+] - [Lac-] - [Cl-]) were lower after training (mean differences) by 2.7 and 1.8 mmol l-1, respectively (P < 0.05). Arterial and femoral venous CO2 tensions and arterial plasma [HCO3-] were lower after training (mean differences) by 1.7 mmHg, 4.5 mmHg and 1.2 mmol l-1, respectively (P < 0.05), with arterial plasma [H+] being greater after training by 2.2 nmol l-1 (P < 0.05). 7. The acute changes in whole blood volume and ion concentrations during maximal exercise were similar to previous observations: Arterial and femoral whole blood [K+] and [Cl-] were increased, whilst [Na+] was lower, across all observation times after training (P < 0.05). 8. Net uptake or release of ions by exercising muscle was assessed by arteriovenous whole blood concentration differences, corrected for fluid movements. A net K+ uptake by muscle occurred at all times, including exercise, but this was not significantly different after training. An increased net Na+ uptake by muscle occurred during exercise (P < 0.05) with greater Na+ uptake after training (P < 0.05). Net muscle Lac- release and Cl- uptake occurred at all times (P < 0.05) and were unchanged by training. 9. Sprint training improved muscle ion regulation, associated with increased intense exercise performance, at the expense of a greater systemic acidosis. Increased muscle Na+ and K+ uptake by muscle during the final seconds of exercise after training are consistent with a greater activation of the muscle Na(+) - K+ pump, reduced cellular K+ loss and the observed lesser rate of fatigue. The greater plasma acidosis found after sprint training was caused by a lower arterial plasma [SID] due to lower plasma [K+] and [Na+], and higher plasma [Lac-]. PMID- 9218231 TI - Length of stay as an outcome in an era of managed care. An empirical study. AB - Longer length of stay (LOS) in substance abuse treatment, a standard measure of treatment success, conflicts with pressures from managed care. To maintain LOS as an outcome, we identified, for four modalities, LOS categories such that program completion rates were relatively constant within category and differed among categories. We validated the cutoffs by showing that future utilization over a 2 year period by clients differed by category. Clients in the long-LOS category used the system in a way consistent with more successful treatment. Thus, rather than using increase in LOS as an outcome, one can use increase in the percentage of clients reaching the long-LOS category. Categories were developed and utilization analyzed for discharges from publicly funded Boston treatment programs between 1/92 and 12/94 from the following modalities: short-term residential (5,462 discharges), long-term residential (5,086 discharges), outpatient (13,656 discharges), and detox (19,965 discharges). PMID- 9218229 TI - Enhanced pulmonary and active skeletal muscle gas exchange during intense exercise after sprint training in men. AB - 1. This study investigated the effects of 7 weeks of sprint training on gas exchange across the lungs and active skeletal muscle during and following maximal cycling exercise in eight healthy males. 2. Pulmonary oxygen uptake (VO2) and carbon dioxide output (VCO2) were measured before and after training during incremental exercise (n = 8) and during and in recovery from a maximal 30 s sprint exercise bout by breath-by-breath analysis (n = 6). To determine gas exchange by the exercising leg muscles, brachial arterial and femoral venous blood O2 and CO2 contents and lactate concentration were measured at rest, during the final 10 s of exercise and during 10 min of recovery. 3. Training increased (P < 0.05) the maximal incremental exercise values of ventilation (VE, by 15.7 +/ 7.1%), VCO2 (by 9.3 +/- 2.1%) and VO2 (by 15.0 +/- 4.2%). Sprint exercise peak power (3.9 +/- 1.0% increase) and cumulative 30 s work (11.7 +/- 2.8% increase) were increased and fatigue index was reduced (by -9.2 +/- 1.5%) after training (P < 0.05). The highest VE, VCO2 and VO2 values attained during sprint exercise were not significantly changed after training, but a significant (P < 0.05) training effect indicated increased VE (by 19.2 +/- 7.9%), VCO2 (by 9.3 +/- 2.1%) and VO2 (by 12.7 +/- 6.5%), primarily reflecting elevated post-exercise values after training. 4. Arterial O2 and CO2 contents were lower after training, by respective mean differences of 3.4 and 21.9 ml l-1 (P < 0.05), whereas the arteriovenous O2 and CO2 content differences and the respiratory exchange ratio across the leg were unchanged by training. 5. Arterial whole blood lactate concentration and the net lactate release by exercising muscle were unchanged by training. 6. The greater peak pulmonary VO2 and VCO2 with sprint exercise, the increased maximal incremental values, unchanged arterial blood lactate concentration and greater sprint performance all point strongly towards enhanced gas exchange across the lungs and in active muscles after sprint training. Enhanced aerobic metabolism after sprint training may contribute to reduced fatigability during maximal exercise, whilst greater pulmonary CO2 output may improve acid-base control after training. PMID- 9218232 TI - Naltrexone plus group therapy for the treatment of opiate-abusing health-care professionals. AB - PURPOSE: The success rate of a treatment program tailored to opioid-abusing health-care professionals that included oral naltrexone and group therapy was studied. METHODS: 20 opioid-abusing health professionals were treated over a 5 year-period. Clients received an initial assessment, supervised administration of naltrexone, and weekly attendance at a psychotherapy group for health professionals. Naltrexone was administered for the first several months, then patients continued the program without naltrexone. RESULTS: 18 patients were referred to the program after being caught diverting medication. Two patients came spontaneously. Of the 18 referred patients, 12 had no relapses, and 5 had only one relapse, followed by long-term sobriety. Mean overall duration of naltrexone administration was 8 months, and the mean duration in the program was 1.9 years. 94% of referred clients had long term abstinence, and 66% were working in their profession during the program. CONCLUSIONS: Naltrexone in the setting of a structured program is helpful in the treatment and professional reinstatement of opioid abusing health professionals. PMID- 9218233 TI - Motivating methadone patients to include drug-free significant others in treatment: a behavioral intervention. AB - The present study introduced a novel behavioral approach for encouraging methadone-treated patients to bring drug-free significant other support into treatment. Seventy-five patients referred to high-intensity psychosocial treatment due to chronic drug use were given 3 weeks to identify a drug-free significant other. Patients noncompliant with this intervention were started on a methadone dose taper that was stopped when significant other support was identified. Patients and their significant others were required to attend a significant other group one time per week for a minimum of 6 weeks. Eighty-five percent of the patients brought a drug-free significant other into treatment. Significant others included family members, partners, and friends. Patients who identified significant other support complied with 77% of their scheduled sessions. The results demonstrated that most methadone patients have drug-free support people who are willing to participate in their treatment. These individuals can be utilized to help patients initiate the process of building new drug-free social support networks. PMID- 9218234 TI - Patterns of service use and treatment involvement of methadone maintenance patients. AB - Although methadone maintenance is an effective treatment for opiate addiction, variations in treatment outcome are evident. These variations may be explained in part by the rehabilitative experiences of patients as reflected in their use of collateral services. This study examined service involvement of 409 methadone maintenance patients at four clinics in order to identify the types of services used and the extent to which potentially rehabilitative services were used. Aside from welfare, there was a strikingly low level of service utilization. Even when services were used, the levels of this use were so low as to be virtually ineffective. These findings regarding treatment and social service utilization suggest that there may not be any attempt to match service provision with patient needs for services. A more rational approach to matching patient needs and available services is thus called for. PMID- 9218235 TI - Personal construct psychotherapy of addictions. AB - Personal construct psychotherapy and its utility in understanding and treating addictions is explored. Several clinical phenomena related to chemical dependency are discussed from the perspective of constructivism. Framed within the context of Prochaska, DiClemente, & Norcross' (1992) stages of change model, several psychotherapeutic techniques are outlined. These concepts and techniques are offered as theoretically based heuristics and are intended to illustrate the potential utility of a clinical approach based upon personal construct theory. PMID- 9218236 TI - Mapping-enhanced drug abuse counseling: urinalysis results in the first year of methadone treatment. AB - Urinalysis (UA) tests for opiates and cocaine were obtained over a 12-month period for a total of 155 long-term clients who participated in treatment in one of three urban methadone centers. At admission, clients were randomly assigned to "node-link mapping" (n = 82) or "standard" (n = 73) counseling treatment. Node link mapping is a strategy for visually representing interrelationships between clients' ideas, feelings, and experiences. These multirelational maps are developed (usually by counselors) during individual and group counseling sessions to clarify clients' issues and problems. The results revealed that (a) mapping clients had significantly fewer opiate-positive UAs during months 2-6 of treatment and (b) session attendance was a significant predictor of cocaine positive UAs over months 2-12 for mapping clients. PMID- 9218237 TI - Facilitation of internal locus of control in adolescent alcoholics through a brief biofeedback-assisted autogenic relaxation training procedure. AB - The purpose of the study was to determine if autogenic relaxation training facilitated through biofeedback promotes an increase in internal levels of locus of control. The participants were residents of two Southwest Missouri alcohol treatment centers and ranged in age from 18 to 21 years. Treatment and control groups were compared on their responses on the Drinking Related Locus of Control Scale (DRIE) and fingertip temperature pre- and posttraining. The training was effective in teaching autogenic relaxation as demonstrated by increased fingertip temperature for the treatment group posttraining, while no differences were observed for the control group. Most importantly, the treatment group was not only significantly more internal in their locus of control after training but were also significantly more internal than the control group posttraining. Given that alcoholics are significantly more external in their locus of control than nonalcoholics, and that an internal locus of control implies an individual's belief that he or she has control and is responsible for his or her behavior, autogenic relaxation facilitated through biofeedback may be a very important component in therapeutic intervention for adolescent alcoholics. PMID- 9218238 TI - Acute dually diagnosed inpatients: the use of self-report symptom severity instruments in persons with depressive disorders and cocaine dependence. AB - A total of 84 adult inpatient dual-diagnosis (cocaine dependence and unipolar depressive disorder) patients' charts were reviewed. Patients were administered the BDI, SCL-90-R with Additional Items for Major Depressive Syndrome, and the DAST-20 within 5 days of admission as standard assessment practice in a hospital program. Depression scores for the BDI and the SCL-90-R were generally consistent, respectively, across each of the depressive disorder diagnostic groups with the exception of organic (cocaine-induced) mood disorder, which had lower mean scores on both instruments. BDI mean score differences were statistically significant regarding the depressive and organic (substance induced) patients. DAST-20 mean scores were consistent across the diagnostic groups (substantial drug abuse range). Results suggest potential discriminating ability of the BDI in particular in distinguishing genuine unipolar depressive disorders from organically (cocaine) induced mood symptomology when assessments are done rapidly following cocaine cessation. PMID- 9218239 TI - Self-disclosure and the treatment of substance abuse. AB - Identification and treatment of the substance-abusing physician has led to outcome studies focusing on years of abstinence and resultant work performance, but little has been written addressing the therapeutic changes recovery brings in the personal lives of these physicians or in their approach to similarly addicted patients. The unique position of recovering psychiatrists engaged in the treatment of substance-abusing patients has not been addressed. Self-disclosure is a major issue and becomes unavoidable if the psychiatrists meet their patients at Twelve-Step meetings. This paper begins a discussion of the issues of self disclosure and anonymity for recovering psychiatrists. The informal method used to gather information highlights the sensitivity of these issues for both patients and psychiatrists. The clinical examples show the importance of evaluating the psychodynamic impact of self-disclosure on the patient, the psychiatrist, and their treatment relationship. PMID- 9218240 TI - Participation in community residential treatment and substance abuse patients' outcomes at discharge. AB - OBJECTIVE: The study sought to identify patient characteristics that predict participation in substance abuse treatment in community residential facilities (CRFs) and to examine the association between patient characteristics, participation in treatment, and outcomes at discharge from CRFs. METHODS: A sample of 2,794 patients with substance abuse disorders was assessed at entry into and discharge from a representative set of 88 CRFs nationwide. RESULTS: In general, patients' psychological distress, motivation for treatment, prior involvement in self-help, and social resources predicted more engagement in CRF services and activities; prior inpatient treatment and the history of a psychiatric disorder predicted less engagement. These patient characteristics also predicted outcomes at discharge; more important, participation in treatment was positively and independently associated with such discharge outcomes as completion of the program and moving into stable residence. In addition, there was some evidence that participation in treatment counteracted the negative effects of high-risk patient characteristics on outcome. CONCLUSIONS: Participation in treatment is as important a predictor of outcomes at discharge from CRFs as are patient characteristics at intake to treatment. Suggestions are made about how providers can enhance patients' motivation to participate and remain in treatment. PMID- 9218241 TI - Controlled drinking as a treatment goal in Australian alcohol treatment agencies. AB - Under the broad umbrella of harm minimisation, the Australian National Drug Strategy has emphasised the development of services aimed at reducing hazardous alcohol consumption in problem drinkers thereby shifting the focus of treatment from abstinence to moderation goals. The objective of the present study was to determine the extent to which alcohol treatment agencies offered advice and treatment aimed at moderation of alcohol consumption (i.e., controlled drinking). Of the 179 agencies (40% of identified treatment agencies across Australia) approached, 66% (115) reported giving advice about controlled drinking as a treatment goal. The reported therapeutic strategies used to assist in the attainment of a controlled drinking goal are empirically supported. Thus, controlled drinking as a treatment goal is widely offered by Australian treatment agencies, by workers who appear well versed in validated strategies and techniques used to obtain such a goal. This finding is discussed in relation to comparison studies conducted in the UK and the USA. PMID- 9218243 TI - Interaction of eye-, head-, and trunk-bound information in spatial perception and control. AB - This article reviews the author's investigations on the perception and control of spatial relations if the carriers of the relevant sense organs are mobile and controlled independently of each other. In the dragonfly, head rotation is controlled by the head's inertia, as well as by cervicocollic, optokinetic, and dorsal light reflexes and, in turn, controls trunk rotation by means of neck reflexes on the wings. In humans, invariance of head-referenced visual direction under eye-to-head rotation is attained by feedforward of an efference copy. In the pigeon, invariance of responses to trunk tilt under head-to-trunk rotation is, in flight, achieved by feedforward of head-to-trunk information yielded by neck receptors. But in standing or walking, this is accomplished by means of gravity sense organs in the trunk. Such organs are also shown to exist in the human trunk by means of experiments on a sled centrifuge. From tests with paraplegic and neuromectomized subjects, it is concluded that truncal graviception 1) is not influenced by mechanoreceptors in the legs, the skin, and between the vertebrae, but 2) is affected by at least two afferent inputs, one originating in the kidneys, another in the tissues that support the large blood vessels against the gravitational load. These conclusions are corroborated by experiments with bilaterally nephrectomized subjects and by means of positive air pressure to the legs, respectively. Recent results under application of positive and negative air pressure to the entire lower body indicate that yet another source of somatic graviception may exist, for example, one that exploits the hydrostatics of blood pressure or the inertia of the mass of the abdominal viscera. PMID- 9218242 TI - Afrocentric treatment in residential substance abuse care. The Iwo San. AB - Alcohol and other drug treatment programs continue to report relatively low success rates among African-American participants. We propose that there is a need to consider treatment approaches that are more culturally competent. An Afrocentric paradigm is suggested and instituted as the central theme of a residential drug treatment program. Elements of an Afrocentric orientation and how these principles are used to guide the development of a treatment philosophy are discussed. PMID- 9218245 TI - Interactions within and between the spatial senses. AB - This paper reviews five types of interaction between sources of spatial information within and between sense organs; 1) nested, 2) opponent, 3) comparison, 4) covariation, and 5) multicue interactions. Efference copy is treated as a type of sensory input. Examples of each type of interaction are provided, with an emphasis on visual-vestibular interactions. In the first type of interaction, inputs from nested sensory systems are summed like vectors. For instance, the 3-D vector sum of inputs from the joints and muscle spindles of the arm allows one to judge the position of the hand. In the second type, inputs from opponent systems are combined to form a signed difference signal with respect to a norm. For instance, the push-pull linkage between the vestibular organs on the two sides of the head provides the signal for head rotation. The third type involves comparisons based on the detection of differences between stimuli presented to different regions of the same sense organ or to distinct sense organs. The fourth type involves the extraction of products or ratios between stimuli used in the detection of invariant high-level features. For instance, the linear size of an object can be derived from the constant product of the distance of the object and the size of its image. Similar systems are used to scale the response to one stimulus feature with respect to a second feature. For instance, vestibular inputs evoking eye nystagmus are scaled by viewing distance. Judgments based on all of the above mechanisms are relational, meaning that they require information from several sources. The fifth type involves multicue systems in which alternative cues are available for the same judgment. The cues are sometimes combined as a weighted mean. For instance, the direction of an object is derived from the mean position of the images in the two eyes, or a judgment of the rotation of the body may be based on combined inputs from the vestibular system and from visual motion. For distinct types of cue, averaging is less common than cue dominance, dissociation, or cue reinterpretation. PMID- 9218244 TI - The role of reafference in recalibration of limb movement control and locomotion. AB - The reafference model has frequently been used to explain spatial constancy during eye and head movements. We have found that its basic concepts also form part of the information processing necessary for the control and recalibration of reaching movements. Reaching was studied in a novel force environment--a rotating room that creates centripetal forces of the type that could someday substitute for gravity in space flight, and Coriolis forces which are side effects of rotation. We found that inertial, noncontacting Coriolis forces deviate the path and endpoint of reaching movements, a finding that shows the inadequacy of equilibrium position models of movement control. Repeated movements in the rotating room quickly lead to normal movement patterns and to a failure to perceive the perturbing forces. The first movements made after rotation stops, without Coriolis forces present, show mirror-image deviations and evoke perception of a perturbing force even though none is present. These patterns of sensorimotor control and adaptation can largely be explained on the basis of comparisons of efference copy, reafferent muscle spindle, and cutaneous mechanoreceptor signals. We also describe experiments on human locomotion using an apparatus similar to that which Mittelstaedt used to study the optomotor response of the Eristalis fly. These results show that the reafference principle relates as well to the perception of the forces acting on and exerted by the body during voluntary locomotion. PMID- 9218247 TI - Vestibular bibliography. PMID- 9218246 TI - Vestibular-neck interaction and transformation of sensory coordinates. AB - The article considers findings and concepts on vestibular-proprioceptive interaction for self-motion perception and postural control under the form of simple describing models. It points out that vestibular-neck interaction is only a small fraction of an extended mechanism of co-ordinate transformations. This links together the different parts of our bodies, so that sensory information arising in one part of the body can be used for perceptual or motor tasks in other parts. Particular emphasis is put on the problems that arise from imperfect signal transduction in the vestibular semicircular canal systems at low stimulus frequencies/velocities. Also, a "down-and-up-channeling" principle is suggested, by which the body support is linked via coordinate transformations to the internal notion of physical space provided by the vestibular system. Furthermore, the following question is addressed: how does the brain use visual input to overcome the vestibular deficiencies, at the risk of visual self-motion illusions? Finally, a conceptual model of postural control is presented in which a proprioceptive feedback loop that links the body to its support surface is merged with a loop for postural stabilization in space. PMID- 9218248 TI - Receptor biology of the melanocortins, a family of neuroimmunomodulatory peptides. AB - Melanocortins, melanocyte-stimulating hormones (MSH) and adrenocorticotropic hormone (ACTH) are homologous natural peptides derived from pro-opiomelanocortin (POMC). Recent breakthroughs in melanocortin receptor (MCR) biology are relevant to neuroimmunomodulation because melanocortins are known to modulate fever, inflammation and immunity, by acting both on peripheral targets and within the brain. During fever, endogenous melanocortins exert antipyretic effects by acting on MCR located within the brain, suggesting a protective counterregulatory role of the central melanocortin system. MCR are also found in melanocytic cells and adrenal cortical cells, the classical targets for alpha-MSH and ACTH, respectively, in myelogenous and lymphoid tissues, and in various endocrine and exocrine glands, adipocytes, and in autonomic ganglia. In the CNS, MCR are prominently distributed in close proximity to the terminal fields of melanocortinergic neurons that innervate neuroendocrine and autonomic motor nuclei as well as other subcortical brain regions important in neuroendocrine and autonomic regulation, sensory processing and various aspects of behavior. Furthermore, the presence of MCR in circumventricular organs of the brain provides direct access of systemic melanocortin hormones to central MCR. Together, these attributes provide an anatomical basis for bidirectional MCR mediated communication between brain and periphery. A group of five G-protein associated MCR subtypes, each of which is positively coupled to adenylate cyclase, has been identified. Among these, the adrenal ACTH receptor (MC2-R) is selectively activated by ACTH. In contrast, the other MCR subtypes (MC1-R, MC3-R, MC4-R, MC5-R) recognize a common group of ligands that includes various forms of MSH as well as ACTH; nevertheless they do exhibit important differences in ligand selectivity. MCR concentrations and MCR mRNA levels are influenced by availability of cognate ligands, by drugs, and by pathological stimuli. Two types of endogenous MCR antagonist proteins have been discovered: agouti protein and the corticostatins. Agouti protein dramatically alters coat color in mammals by antagonizing melanocytic MC1-R. Moreover, spontaneous dominant mutations of the agouti gene in several strains of mice lead to its ubiquitous overexpression and produces not only yellow coat color, but also obesity and insulin resistance, perhaps as a result of its antagonism of other MCR subtypes. The recent emergence of synthetic MCR antagonists, and the feasibility of molecular approaches for targeted inactivation of individual MCR subtypes, should facilitate elucidation of the roles and mechanisms of neuroimmunomodulation by endogenous melanocortins, and the determination of whether selective pharmacological targeting of MCR may ultimately have therapeutic utility. PMID- 9218249 TI - DHEAS inhibits TNF production in monocytes, astrocytes and microglial cells. AB - We previously reported that neurosteroids, including dehydroepiandrosterone sulfate (DHEAS), inhibit the production of TNF in vitro and in vivo. In this paper we evaluated the effect of DHEAS on TNF production by cultured rat astrocytes and murine glial cell clones, and compared it with the effect on monocytic THP-1 cells. We found that DHEAS at a concentration of 10(-4)-10(-7) M inhibits TNF production induced by lipopolysaccharide (LPS, 1 microgram/ml) in these cells. Since the inhibitory effect of DHEAS is not mediated by the glucocorticoid (GC) receptor and DHEAS is an allosteric antagonist of the GABAA receptor, we investigated the possible role of GABAA receptors in this effect. The results showed that the inhibitory effect of DHEAS (10(-6) M) on TNF production by THP-1 cells was completely reversed by addition of 10(-6) M GABA. However, a GABAA receptor antagonist (bicuculline) did not mimic the action of DHEAS. In conclusion, DHEAS can inhibit TNF production in astrocytic and microglial cells suggesting it could be an endogenous regulator of TNF production in the brain. PMID- 9218250 TI - Neuronal expression of tumor necrosis factor alpha in the murine brain. AB - We have examined the expression of the inflammatory cytokine, tumor necrosis factor alpha (TNF alpha) in the mouse brain. Using immunohistochemical methods developed, we found anti-TNF alpha immunoreactivity localized in the basal ganglia and other discrete brain structures. Constitutive immunoreactivity, present in normal, unstimulated brain, was observed in glial and microglial-like cells, but it was predominant in neuronal-like cells. Intravenous administration of lipopolysaccharide (LPS) increased TNF alpha transcript levels detected by RT PCR in specific brain subregions in which contaminating blood cells were removed. The maximal increase occurred within 2 h of LPS injection; transcripts diminished to near control levels in the next 4 h. Immunocytochemical analysis and single cell RT-PCR analysis of primary cultures of cortical neuronal cells confirmed expression of TNF alpha in cells that also express neuronal-specific enolase RNA. Addition of LPS or recombinant TNF alpha protein to neuronal cultures enhanced expression of TNF alpha transcripts. Our results indicate that in addition to glial and microglial cells, a well-defined subset of neuronal cells also express TNF alpha constitutively; this expression can be altered by both extrinsic (LPS) and intrinsic (TNF alpha itself) factors. PMID- 9218252 TI - [IC-IC bypass]. PMID- 9218251 TI - Interleukin-1 receptor accessory protein transcripts in the brain and spleen: kinetics after peripheral administration of bacterial lipopolysaccharide in mice. AB - Receptors for IL-1, type I (IL-1R1) and type II (IL-1R2), have been characterized by pharmacological and molecular techniques in the mouse brain. High densities are mainly found in the cortex, dentate gyrus and choroid plexus. It was therefore of interest to investigate the expression of mRNA IL-1 receptor accessory protein (IL-1R AcP), which is a part of the IL-1 receptor complex and has been shown to interact specifically with IL-1R1. IL-1R AcP transcripts were detected under basal conditions following RT-PCR amplification in the mouse brain, as well as in the pituitary, spleen, adrenal and liver. IL-1R AcP transcripts were found in higher amounts than IL-1R1 transcripts in all tissues except the spleen, where their expression was minor. Following bacterial lipopolysaccharide (LPS) stimulation (3-48 h), IL-1R AcP transcripts were not changed in the brain, while IL-1R1 transcripts were increased for 3-6 h. In the spleen, a slight increase in IL-1R AcP and IL-1R1 was observed during the first hours following LPS stimulation. In conclusion, IL-1R AcP mRNA is expressed in the brain and in other tissues where IL-1R1 transcripts are found. However, the regulation of its expression is distinct from IL-1R1. The high level of expression and the lack of regulation of IL-1R AcP transcripts in the brain under inflammatory conditions suggest that the protein might be constitutively expressed in excess. PMID- 9218253 TI - [Efficacy of the TURBO-fluid attenuated inversion recovery spin echo sequence of MRI as a preoperative neuroradiological examination]. AB - Whenever the extirpation of intracranial tumors is planned, neurosurgeons always keep their eyes on the cerebrospinal fluid (CSF) space around intracranial tumors. If enough space exists in the neighborhood of the tumors, the damage to adjacent parenchyma may be reduced by the procedure through the CSF space. A newly advanced MRI pulse sequence: the FLAIR (fluid attenuated inversion recovery) imaging, in which a long TE spin echo sequence is used with suppression of the CSF with an inversion pulse, displays the CSF space as a no-signal intensity area. There have been only a few reports, however, on the FLAIR pulse sequence of brain tumors as yet. We examined 34 cases of intracranial tumors by FLAIR images and analyzed the advantages and disadvantages of the FLAIR pulse sequence for decision making on tumor removal. Making use of the FLAIR pulse sequence, the CSF space is depicted as a no-signal intensity area and much more information about perifocal edema and the invasion area around the tumors can be provided than that provided by the other ordinary pulse sequences (T1 weighted images, T2 weighted images and Proton weighted images). Therefore, operative strategies can be more easily worked out on the FLAIR images. Furthermore, the difference between arachnoid and epidermoid is able to be detected on the FLAIR images. Nevertheless, on FLAIR images, the tumors without perifocal edema or invasion to adjacent parenchyma were not apparent and the difference between tumoral dissemination into multi-ventricular space and the periventricular artifact of FLAIR images could not be distinguished. The FLAIR pulse sequence has other artifacts like intraventricular flow related enhancement and so on. If the images are carefully checked up on the above-mentioned points, the FLAIR pulse sequence of MRI can not fail to be useful in making plans for operations on intracranial neoplasms. PMID- 9218254 TI - [Clinicopathological studies of craniocerebral gunshot injuries]. AB - Gunshot wounds are rare in Japan because of few regulatory laws against the possession of guns. Nevertheless such wounds are increasing in prevalence these days. Reports on the microscopic findings concerning these intracerebral lesions are fewer than those on the macroscopic findings in the scalp, the skull and the intracranial cavity. In this study we evaluated computed tomographical and histopathological findings in craniocerebral gunshot injuries. CASES: Nine patients with gunshot wounds to the head were presented. All were male and the age ranged from 17 to 66 years. Four were suicides and four were attempted murders and the last one was of unknown etiology. Morphological examination was performed on 5 autopsy cases. The distance of the bullet from the cranial cavity was as follows: long distance, 4 cases; close contiguity, 5 cases. The calibers of the weapons were as follows: 38 mm in 6 cases, 45 mm in 1 case and unknown in 2 cases. RESULTS: CT scans were examined in six cases, which revealed a missile track, hemorrhagic contusion, traumatic subarachnoid hemorrhage and marked tension pneumocephalus. In some cases, CT scan also revealed bony and metallic fragments, some deep within the cranial cavity. In the histopathological study, we found marked swollen brain (brain weight over 1500 mg) and hemorrhagic contusion in the vicinity of the missile track and interhemispheric fissure, and widespread traumatic subarachnoid hemorrhage and intraventricular hematoma. We would like to emphasize especially the remote contusion seen in the distant part of the missile track as well as massive exsudation and hemorrhage around the nerve fiber bundles. Remote contusion was observed in the inferior surface of the fronto-temporal lobes, and bilateral hemorrhagic contusion was seen in the vicinity of the superior longitudinal fissure on CT scans and autopsy findings. In one case, the bullet rotated within the intracranial cavity. In conclusion, nine cases of craniocerebral gunshot injuries were examined, while we also reviewed the medical literature concerning the shearing injury produced by gunshot brain wounds. The head injuries were further delineated by the correlation between autopsy and computerized tomography findings. PMID- 9218255 TI - [Preservation of olfaction in frontal transbasal approach]. AB - Lesions in the frontal base and clival area have conventionally been approached using the transbasal approach described by Derome and Guiot. However, this approach necessitates removal of the crista galli and sectioning of the olfactory rootlets with the associated risk of anosmia and cerebrospinal fluid leak and, in addition, complex reconstruction of the frontal base is required. We describe a new approach to deeply situated tumors in the frontal base, parasellar and clival area which is a modification of Spetzler's craniofacial approach with preservation of olfaction. In this approach, circumferential osteotomy cuts are made around the cribriform plate to permit en block removal with its attachment to both the dura and underlying mucosa. Opening of the dura is avoided and the cribriform bone is used to reconstruct the frontal base. Three patients underwent surgery using this approach for treatment of recurrent pituitary adenoma in two cases and for clivus chordoma in one. In one patient, olfactory function was not preserved because resection of nasal mucosa was small. In the other two patients, however, olfaction was preserved by creating a cribriform plate complex with a sufficient area of resection of nasal mucosa and tumors were completely removed. Olfaction can be preserved, CSF leakage can be prevented, and facial skin incision and complex frontal base reconstruction can be avoided when this technique for maintaining normal olfactory-cribriform anatomy is used in frontal transbasal approaches. PMID- 9218256 TI - [Dural arteriovenous fistula involving the superior sagittal and transverse sigmoid sinuses, treated by thrombolysis: case report]. AB - A rare case of dural arteriovenous fistula (DAVF) in the superior sagittal sinus (SSS), the transverse sinus and the sigmoid sinus is reported. A 64-year-old man, who had had an episode of temporary visual disturbance after moderate fever for a week about 20 years before, was aware of loss of visual acuity and reduced field of view in the right eye. When he was introduced to our outpatient service, increased intracranial pressure (ICP) was detected by lumbar puncture. Cerebral angiograms showed bilateral DAVFs both in the posterior fossa and the SSS concomitant with thrombosis in the transverse sinus, sigmoid sinus and SSS. Afterwards, endovascular transarterial embolization through bilateral occipital, posterior auricular and left middle meningeal, superior temporal arteries was carried out. In addition, transvenous thrombolytic therapy using a catheter inserted into SSS resulted in the improvement of his visual problems. Although he was discharged at once, he was readmitted to our department with Foster Kennedy syndrome and increased ICP. Cerebral angiograms showed recurrence of both DAVF and sinus thrombosis. Transarterial embolization was performed again resulting in a significant reduction of DAVF, and his visual acuity was recovered to a moderate degree. The origin of DAVF is still controversial. Although two theories, "congenital" and "acquired", are put forward, it has been thought that both factors play important roles. In our case, the patient had stenosis in the jugular canal portions of the sigmoid sinus. In addition, sinus thrombosis seemed to have occurred. It is thought that increased intrasinus pressure may have lead to communication with surrounding arteries through existing dural vessels. We applied transvenous thrombolytic therapy in this case. Our result suggests that we should consider this therapy for some cases of DAVF. PMID- 9218257 TI - [Laparoscopic retrieval of a dislocated ventriculoperitoneal shunt catheter: report of three cases and a review of the literature]. AB - Described are 3 cases of a disconnected ventriculoperitoneal shunting system that was successfully retrieved by using a laparoscopic procedure, with a review of the literature. All patients had symptoms of increased intracranial pressure. Roentgenograms showed disconnection of a ventriculoperitoneal shunt catheter at the connecting device and its migration into the peritoneal cavity. A laparoscope was introduced into the peritoneal cavity using the double puncture procedure and the catheter was extracted in less than 15 minutes. The use of a laparoscope enabled exploration of the entire space of the cavity without any large laparotomy incision. Furthermore, the laparoscopic procedure also easily enabled introduction of a replaceable ventriculoperitoneal shunt catheter into the appropriate portion in the cavity and confirmed the CSF flow into the cavity. Because catheters which have migrated into the cavity might cause an acute abdomen, it is important that they should be removed as soon as possible. It should be kept in mind, during the procedures of extracting catheters, that the inner absorptive surface of the peritoneal cavity must be preserved as much as possible. In this regard, laparoscopic retrieval of disconnected shunt catheters is a promising method. PMID- 9218258 TI - [A penetrating cranio-facial injury due to a nail-gun accident]. AB - We report a case of a penetrating cranio-facial injury due to a nail-gun accident. An 18-year-old worker was admitted to our hospital as an emergency patient. He was working using a nail-gun when a nail ricocheted off a wall and pierced the right side of his face. Skull X-rays and a CT scan showed that a 9 cm nail had pierced his right frontal lobe through the right maxillary bone via the orbital space. The patient was alert without any neurologically abnormal findings. A small stab wound was recognized on his face. The nail was removed six hours after the injury through a sublabial approach and a fronto-temporal craniotomy. The nail was very tightly fixed in the maxillary bone and skull base bones, and a screw driver normally used in orthopedic surgery proved to be very useful for removing the nail. The patient returned home 14 days later without any neurological deficits. The technical problems associated with such a nail-gun injury in the face and skull are also discussed. PMID- 9218259 TI - [A case of foramen magnum meningioma in which case enhanced three-dimensional CT scan was valuable for preoperative evaluation of the surgical approach]. AB - We report a case of foramen magnum meningioma in which case enhanced three dimensional CT scan was valuable for preoperative evaluation of the surgical approach. A 53-year-old woman had suffered from stiffness and pain in the left occipital region and numbness of the left side of the face for about 2 years before admission. She had also weakness and numbness of the left side of her body for about 2 months before admission, and dysphagia and pain in the occipital region and in the posterior region of the neck produced by straining for about 1 month before admission. Neurological examination revealed left hemiparesis, and hypalgesia and tactile hypesthesia of the left side of the body, including the face. Plain X-P was normal. Enhanced CT scan and gadolinium enhanced MRI revealed a well-enhanced mass attached to the left anterolateral part of the foramen magnum. The left occipital condyle was observed at the lateral side of the attachment part of this mass. Angiography revealed tumor feeders from the meningeal branches of the left vertebral artery and the left ascending pharyngeal artery. Enhanced three-dimensional CT scan clearly showed that the tumor was attached to the left anterolateral part of the foramen magnum, that the left occipital condyle was at the lateral side of the attachment part of this mass and that the jugular foramen and jugular tubercle were situated superolateral to the attachment part of this mass. Considering these factors, we decided that removal of the posterior part of the left occipital condyle was necessary, but removal of the left jugular tubercle was not necessary for a good operative view from the left posterior lateral direction. The tumor was totally removed successfully and good results were obtained by the transcondylar approach without removal of the jugular tubercle. Histology of the tumor revealed meningothelial meningioma. In this case, preoperative evaluation with enhanced three-dimensional CT scan was helpful for deciding the surgical approach. With enhanced three-dimensional CT scan, it is easy to judge whether removal of the posterior part of the occipital condyle and/or the jugular tubercle is necessary for a good operative view, and we can get good images revealing the relationships between the tumor and surrounding structures. Preoperative evaluation with enhanced three-dimensional CT scan is very useful especially in this kind of situation. PMID- 9218260 TI - [Large cystic neurinoma: a case report]. AB - We report a case of large cystic acoustic neurinoma. A 52-year-old male was admitted to hospital with a history of progressive dysphagia, gait disturbance and diplopia for 2 months. On admission, neurological examinations revealed Bruns' type nystagmus to the left side, hypesthesia in the distribution of the second and third divisions of the left trigeminal nerve, and partial paresis of cranial nerves IX, X, and XII on the left side, and truncal ataxia. A pure-tone threshold audiogram indicated the presence of 32 dB hearing loss in the left ear. Speech discrimination was 80%. Caloric vestibular responses were absent on the left side. Skull radiographs with polytomographs of the internal auditory canal (IAC) were normal. Bony changes in the IAC were not found by high-resolution bone window computed tomography (CT) scan. A plain CT scan revealed a large low attenuated cystic mass in the left cerebellopontine angle (CPA), which was associated with displacement of the fourth ventricle. An enhanced CT scan demonstrated a thin rim-enhancement in the cyst wall. Magnetic resonance imaging (MRI) scans disclosed a large rim-enhanced cystic mass extending superiorly into the tentorial incisura and inferiorly into the foramen magnum. At surgery via a left suboccipital approach, a large cystic mass was found at the left CPA arising from the VIIIth nerve, and compressing the Vth, VIth, VIIth and lower cranial nerves. The cyst was filled with a xanthochromic fluid and was firmly attached to the internal auditory meatus (IAM). However no tumor extension into the IAM was confirmed. The tumor was excised completely. The postoperative course was uneventful, except for impairment of the VIIth and VIIIth nerves. At 6 months after the first operation, the facial nerve had improved up to grade III (Hause Brackmann stage). Histological examinations revealed a typical benign acoustic neurinoma with predominant representation of Antoni B tissues. The cyst wall contained numerous abnormal sinusoid and telangiectasia-like vessels which showed occasional thromboses. The vessel walls displayed endothelial proliferations and were frequently hyalinized. Hemosiderin deposits and hemosiderin-containing phagocytes were also found near these vessels. Myxoid degeneration and necrosis were evident in vast areas. These degenerative changes appeared to be the principal causes of the large cystic formation. 16 cases including our case have been reported. The broad characteristics of the clinical symptoms and radiological findings of these tumors are discussed. PMID- 9218261 TI - [A case of malignant lymphoma after renal transplantation]. AB - A case of a primary brain malignant lymphoma after renal transplantation and immunosuppressive therapy is reported. A 41-year-old male patient had been treated with 125 mg/day of azathioprine and 10 mg/day of prednisolone after renal transplantation. He had also been suffering from various infectious diseases. Multiple brain tumors were found and diagnosed as having B-cell, diffuse large cell type malignant lymphoma. In spite of moderate response to irradiation, he died of pneumonia. The anti-Epstein Barr virus antibodies changed from a negative to a positive level after renal transplantation and they increased markedly after brain malignant lymphoma had been found. The number of T- and B-lymphocytes also decreased markedly at that time. So the Epstein-Barr virus was suspected to be the cause of the malignant lymphoma. PMID- 9218262 TI - [Transient decrease in the size of an enhanced anaplastic astrocytoma seen on magnetic resonance imaging: a case report]. AB - We report unusual radiographic findings which were seen during the management of a patient with anaplastic astrocytoma. An enhanced region in a gyrus of the right frontal lobe was demonstrated in a 38-year-old woman who had had a generalized seizure. Following treatment with steroid- and osmotherapy, this enhanced region decreased clearly on magnetic resonance imaging (MRI). Six months later, an enhancing mass lesion appeared in the same position. After surgery, this was diagnosed as being an anaplastic astrocytoma. It is speculated that the initial enhancement was caused by transient dysfunction of the blood-brain following the seizure. In this case, the most important radiologic image was a T2-weighted image of MRI which was able to demonstrate the existence of the lesion until the time of its removal by surgery. PMID- 9218263 TI - [A malignant rhabdoid tumor appearing simultaneously in the kidney and the brain of an infant: case report]. AB - A 6-month-old female was admitted to the hospital with bad temper and decreased sucking power. CT scans revealed tumors in her right kidney and left cerebellum. The patient underwent right radical nephrectomy to excise the kidney tumor. The pathological diagnosis was malignant rhabdoid tumor (MRT). Seven days later, the patient underwent left suboccipital craniectomy for total excision of the cerebellar tumor. The cerebellar tumor existed extraaxially, and consisted of a solid mass lesion and a cystic lesion. Histological examination revealed that it was also a malignant rhabdoid tumor. A follow-up CT, 1.5 months after surgery, revealed a recurrence of the kidney tumor and metastasis to the chest wall and lung. The patient received 16.9 Gy radiotherapy to the abdominal tumor and chemotherapy with etoposide, carboplatin, and ifosfamide. However, she died of respiratory insufficiency 4 months after surgery, though neither recurrence nor metastasis was found in the brain. Nor was there evidence of leptomeningeal dissemination. MRT is a highly malignant tumor that occurs most frequently in the kidney. However, it can also occur in other tissues, including the brain. This tumor occurs most commonly in children under 2 years of age. There is a 3:2 male predominance. The median length of overall survival of MRT in the brain is 6 months. MRT contains nests or sheets of rhabdoid cells. A typical rhabdoid cell has an eccentric round nucleus with a prominent nucleolus and a plump cell body. MRT is composed entirely or partly of rhabdoid cells. Many MRTs have other components, such as PNET areas, mesenchymal area, and epithelial areas. For this reason, they are sometimes called atypical teratoid/rhabdoid tumors. MRTs in the brain contain fewer rhabdoid cell areas than MRT in the kidney. This makes diagnosing MRT in the brain more difficult. A careful search of the entire specimen for variations in pattern and cell type, along with application of immunohistochemical methods is the most useful method of obtaining a diagnosis. In our case, the cerebellar tumor consisted of rhabdoid cell areas, mesenchymal areas, and PNET areas. The cerebellar tumor contained fewer rhabdoid cell areas than the kidney tumor. However, the rhabdoid cell areas in the cerebellar tumor were almost the same as those in the kidney tumor. Furthermore, immunohistochemical staining was positive for vimentin and keratin in the rhabdoid cell areas. Therefore, we were able to make a diagnosis of MRT. It is possible that some of the previously reported cases diagnosed as CNS PNET were actually MRT in the brain, especially if the cases were associated with MRT in the kidney. PMID- 9218264 TI - Intravenous cocaine self-administration in mice lacking 5-HT1B receptors. AB - The present experiment tested the hypothesis that 5-HT1B receptors are involved in the reinforcing effects of cocaine. Transgenic mice lacking 5-HT1B receptors were used as subjects and compared with wild-type mice for the acquisition and maintenance of intravenous (IV) cocaine self-administration. Male 129/Sv-ter and 5-HT1B-minus 129/Sv-ter inbred mice (Columbia University, New York) were initially trained to press a lever under a fixed-ratio schedule 2, first for sweetened condensed milk as reinforcer and subsequently for cocaine (2.0 mg/kg/infusion). When a stable baseline of responding was obtained, each subject was tested under different doses of cocaine (1.0, 2.0, and 4.0 mg/kg), with the number of reinforcers per hour used as the dependent variable. Both strains successfully acquired food-shaping and cocaine self-administration, but the mutant mice presented a significantly shorter latency to meet IV cocaine self administration acquisition criteria (p < 0.05). However, both wild-type and mutant mice had similar dose-response to cocaine. These results suggest that the 5-HT1B receptors may be implicated in the propensity to self-administer cocaine, but other mechanisms might be involved in the maintenance of cocaine self administration. PMID- 9218265 TI - Modeling drug dependence behaviors for animal and human studies. AB - Laboratory models are available to study drug reinforcement in animals and humans, but few are available to study other drug dependence phenomena, e.g., difficulty stopping or use despite harm. The present paper is a first attempt to illustrate the feasibility of developing such models for use in both nonhuman and human research and discusses their possible utility in research to understand and treat stimulant and other drug dependencies. PMID- 9218266 TI - The influence of alternative reinforcers on cocaine use and abuse: a brief review. AB - This report reviews experimental studies conducted with nonhuman and human subjects demonstrating that: a) cocaine's abuse liability is, in part, a function of its positive reinforcing effects, b) cocaine use is operant behavior, c) the degree of behavioral control that cocaine exerts as a reinforcer is malleable and dependent on environmental context, and d) increasing the availability of alternative, nondrug reinforcers can significantly disrupt the acquisition and maintenance of cocaine use and abuse. Implications of these observations for effective prevention and treatment of cocaine abuse are discussed. PMID- 9218267 TI - Differences in the liability to self-administer intravenous cocaine between C57BL/6 x SJL and BALB/cByJ mice. AB - Application of animal models of psychostimulant abuse for experimentation in mice is becoming increasingly important for studying the contribution of genetic differences, as well as the roles of selected (targeted) genes, in specific behaviors. The purpose of this study was to investigate strain differences in cocaine self-administration behavior between C57BL/6 x SJL hybrid mice and BALB/cByJ mice. These two strains were chosen because BALB/cByJ mice have a well developed behavioral pharmacological profile, and hybrid strains on a C57BL/6 background are commonly used for generating transgenic expressing and knockout mutant mice. C57BL/6 x SJL mice dose-dependently acquired cocaine self administration (1.0 mg/kg/injection but not 0.25 mg/kg/injection) by responding selectively in the active nose-poke hole and maintaining stable levels of daily drug intake; they also exhibited a characteristic inverted-U-shaped cocaine dose effect function. BALB/cByJ mice failed to acquire cocaine self-administration at either dose under the same test conditions. The strain differences observed in self-administration did not seem to be attributed to other behavioral differences because the two strains exhibited similar amounts of spontaneous nose-poking in the absence of reinforcers, and BALB/cByJ mice responded more than C57BL/6 x SJL mice in a food-reinforced nose-poke operant task. Importantly, the dose-effect function for the motor stimulating effects of cocaine (3.8-30 mg/kg intraperitoneally) suggests enhanced sensitivity but reduced efficacy of cocaine in stimulating motor activity in BALB/cByJ mice relative to the C57BL/6 x SJL hybrid mice. These results indicate that the decreased liability of BALB/cByJ mice to acquire cocaine self-administration is not the result of differences in spontaneous activity or performance, but may reflect different sensitivities to the reinforcing, or rate-disrupting, properties of cocaine. The data support an influence of genetic background in the liability to self-administer cocaine. Thus, a hypothesis is proposed that the decreased liability of BALB/cByJ mice to acquire cocaine self-administration is related to differences in brain monoamine systems linked to the high "emotionality" profile of BALB/c mice in novel or fearful situations, including perhaps cocaine administration. PMID- 9218268 TI - A critique of fixed and progressive ratio schedules used to examine the neural substrates of drug reinforcement. AB - This paper is a critique of fixed and progressive ratio schedules used to examine the neural substrates of cocaine reinforcement. The discussion focuses on problems encountered while examining the effects of neurotoxic lesions and pharmacological pretreatments on cocaine reinforcement. We review the theoretical and interpretational problems associated with the use of the fixed ratio (FR) schedules that have been used in the majority of studies, and we conclude that rate of drug intake cannot directly address the issue of increased or decreased reinforcer efficacy. The progressive ratio (PR) schedule offers some advantages over FR schedules, although it is now clear that the same implementation cannot be applied across all drug classes. It is likely that the motivation to self administer psychostimulant vs. opiate drugs is qualitatively different. We conclude that there is no single schedule that can quantify all aspects of drug reinforcement and that behavioral paradigms will need to be adapted according to the particular question under study. PMID- 9218269 TI - Sensitization to the behavioral effects of cocaine: modulation by dynorphin and kappa-opioid receptor agonists. AB - Several lines of evidence suggest an involvement of the mesolimbic dopamine (DA) system in the mediation of psychostimulant-induced sensitization. It is also apparent that endogenous opioid peptide systems can modulate the activity of this same DA system. Psychostimulant-induced alterations in opioid peptide gene expression have also been reported. In this review, evidence will be presented that demonstrates that the administration of kappa-opioid agonists can prevent the initiation of behavioral sensitization to cocaine and that such treatment is also effective in preventing alterations in mesolimbic DA neurotransmission that occur as a consequence of repeated cocaine administration. The putative role of opioid-DA interactions in the modulation of psychostimulant-induced sensitization will also be discussed. PMID- 9218270 TI - Psychostimulant abuse: the case for combined behavioral and pharmacological treatments. AB - Behavioral and pharmacological therapies have been used alone and in combination for the treatment of substance abuse; however, to date, no single treatment approach for psychostimulant abuse has demonstrated widespread efficacy. This paper describes the various functions that are served by both behavioral therapies and pharmacotherapies and their respective mechanisms of action. It is argued that combined treatments can be expected to produce additive effects because the two approaches operate through different and potentially complementary mechanisms. Illustrations of these underlying principles and experimental support for the use of combined treatments are drawn from smoking cessation research, which has broadly applied combined behavioral and pharmacological therapies for treating abuse of nicotine, a mild stimulant. In addition, the results of recent studies that have evaluated the efficacy of behavioral techniques and/or potential pharmacotherapies for treating cocaine abuse are reviewed. Finally, methodological strategies are recommended for future evaluations of combined therapy approaches to conclusively evaluate separate and combined efficacy of treatments for psychostimulant abuse. PMID- 9218271 TI - Long-term cocaine self-administration decreases striatal preproenkephalin mRNA in rhesus monkeys. AB - The consequences of long-term exposure to cocaine on striatal preproenkephalin mRNA were assessed using quantitative in situ hybridization in monkeys that self administered cocaine for approximately 2 years. Autoradiograms revealed high levels of preproenkephalin hybridization signal in both the caudate and putamen of the dorsal striatum, as well as in the shell and core subdivisions of the nucleus accumbens of drug-naive control monkeys. In general, there was a medial to lateral and ventral to dorsal gradient of preproenkephalin mRNA observed within the striatum of normal controls. Preproenkephalin mRNA was significantly reduced in widespread portions of both the dorsal and ventral striatum following chronic long-term cocaine self-administration when compared with levels in normal controls. These data confirm those observed in human drug abusers and suggest that long-term abuse of cocaine can result in significant alterations in the opioid regulation of striatal efferent neurons. PMID- 9218272 TI - Use of positron emission tomography to study the dynamics of psychostimulant induced dopamine release. AB - Microdialysis studies have shown that psychostimulants act through a common neurochemical mechanism of elevating synaptic dopamine content in the mesocorticolimbic dopaminergic system. However, little information is available regarding the dynamics of the interaction between the elevated synaptic dopamine levels induced by a psychostimulant and postsynaptic dopamine receptors. The goal of the current investigation was to determine if positron emission tomography (PET) studies using the dopamine D2-selective radioligand [18F]4' fluoroclebopride ([18F]FCP) could be used to measure synaptic dopamine levels. Rhesus monkeys were used because our previous studies revealed that [18F]FCP has a low test/retest variability in this species. Under control conditions, [18F]FCP had a high uptake and slow rate of washout from the basal ganglia, a region of brain that expresses a high density of D2 receptors, reaching kinetic equilibrium at approximately 40 min. Challenge studies, each separated by at least 1 month, were conducted by administering an intravenous dose of (-)cocaine, d-amphetamine, methylphenidate, or d-methamphetamine (1.0 mg/kg) at 40 min post-IV injection of a no-carrier-added dose of [18F]FCP. In each case, the psychostimulant caused an increase in the rate of washout of [18F]FCP from the basal ganglia. Methamphetamine and amphetamine had more pronounced effects on the washout kinetics of [18F]FCP relative to cocaine and methylphenidate, a result that is consistent with the ability of each drug to elevate synaptic dopamine levels. Our results indicate that challenge studies with [18F]FCP may be a useful technique for studying the dynamics of the interaction between psychostimulant-induced increases in synaptic dopamine and postsynaptic D2 receptors. PMID- 9218273 TI - Behavioral sensitization to ethanol: genetics and the effects of stress. AB - Some aspects of drug abuse syndromes may be influenced by sensitization to some drug effects. This enhancement of drug effect has been associated with prior drug exposure and with exposure to stressful stimuli. It has been postulated that sensitization to psychomotor stimulant drug effects influences sensitivity to drug reward. The drugs of abuse best characterized for sensitization phenomena include cocaine, amphetamine, and morphine. In general, ethanol's molecular mechanisms of action have been difficult to define relative to drugs with known receptor or transporter binding sites and, likewise, ethanol sensitization has been less thoroughly examined. Evidence supporting the existence of behavioral sensitization to ethanol, for genetic differences in the occurrence of ethanol sensitization, and for the influence of corticosterone on the development of ethanol sensitization is reviewed herein. There appear to be different genetic determinants of acute drug sensitivity and sensitization. Cross-sensitization between stress and ethanol suggest a potential role for hypothalamic-pituitary adrenal (HPA) axis associated changes in ethanol sensitization, consistent with mechanisms likely contributing to sensitization to other abused drugs. Furthermore, glucocorticoid receptors appear to mediate both ethanol- and stress induced sensitization to ethanol. A biological link between drug reward and drug sensitization involving HPA axis hormones may exist and, thus, study of the sensitization process may elucidate mechanisms relevant to drug abuse. PMID- 9218274 TI - Cellular mechanisms underlying reinforcement-related processing in the nucleus accumbens: electrophysiological studies in behaving animals. AB - Numerous investigations have implicated the nucleus accumbens (NA) as an important neural substrate involved in mediating reinforcement-related processing. Electrophysiological studies in behaving animals enable a direct examination of cellular mechanisms underlying this process via characterization of NA activity at critical times during responding for food, water, or drug reward. Electrophysiological studies are reported that examined the activity of NA neurons during water- and cocaine-reinforced responding in rats. These studies reveal that some NA neurons exhibit changes (increases or decreases) in firing rate synchronized to the response-contingent delivery of water or cocaine. Furthermore, the sampled population of NA neurons exhibited less synchronized cell firing during the response for cocaine than for the water reward. NA activity during cocaine self-administration was explicitly coupled to the behavioral state of the animal and was markedly influenced by the stimulus context in which the drug was delivered. These findings are discussed with respect to the dynamic properties of NA activity and its importance as an underlying cellular substrate mediating reinforcement-related events in the behaving animal. PMID- 9218275 TI - Relations between heterogeneity of dopamine transporter binding and function and the behavioral pharmacology of cocaine. AB - Both in vitro binding studies and studies of dopamine uptake have indicated that there is a heterogeneity of action of cocaine and cocaine analogs. Both high- and low-affinity binding sites have been identified. Some drugs that bind to the dopamine transporter show both high- and low-affinity components whereas others do not. Behavioral studies have indicated that the high-affinity component appears to be the one most directly involved in the actions of cocaine related to abuse. These conclusions are based on correlations of affinities and psychomotor stimulant effects. In addition, tolerance to the psychomotor stimulant effects of cocaine occurs with a concomitant change in only the high-affinity component for dopamine uptake. Certain dopamine uptake inhibitors may have only actions mediated by the low-affinity component. These drugs bind to the dopamine transporter and inhibit dopamine uptake; however, they do not have behavioral effects like those of cocaine. This finding is a critical point of inquiry for the dopamine hypothesis because, based on the neurochemical data, these drugs should have behavioral actions like those of cocaine. In contrast, some of these drugs antagonize the behavioral effects of cocaine, suggesting that the low affinity site somehow modulates the actions mediated by the high-affinity site. Recently, some benztropine analogs have been discovered that bind to the dopamine transporter and inhibit dopamine uptake monophasically but have behavioral effects that are dissimilar to those of cocaine. These compounds may prove useful in determining the behavioral significance of heterogeneity of actions at the dopamine transporter. Further, these studies may provide leads to novel therapeutics for the treatment of cocaine abuse. PMID- 9218276 TI - Opponent process model and psychostimulant addiction. AB - There are many sources of reinforcement in the spectrum of cocaine dependence that contribute to the compulsive cocaine self-administration or loss of control of cocaine intake that constitutes the core of modern definitions of dependence. The development of withdrawal has long been considered an integral part of drug addiction but has lost its impact in the theorization of drug dependence because of new emphasis on the neurobiological substrates for the positive-reinforcing properties of drugs. The present treatise reviews the neurobiological substrates for the acute positive reinforcing effects of cocaine and what is beginning to be known about the neurobiological substrates of cocaine withdrawal. The concept of motivational or affective withdrawal is reintroduced, which reemphasizes opponent process theory as a model for the motivational effects of cocaine dependence. The same neural substrates hypothesized to be involved in the acute reinforcing properties of drugs (basal forebrain regions of nucleus accumbens and amygdala) are hypothesized to be altered during chronic drug treatment to produce the negative motivational states characterizing drug withdrawal. Within these brain regions, both the neurochemical system(s) on which the drug has its primary actions and other neurochemical systems may undergo adaptations to chronic presence of the drug. An understanding of the adaptations of the motivational systems of the brain accompanying cocaine dependence leads to important predictions not only about the etiology, treatment, and prevention of cocaine addiction but also about the vulnerability of these motivational systems in non drug-induced psychopathology. PMID- 9218277 TI - Dose-effect functions for cocaine self-administration: effects of schedule and dosing procedure. AB - Research related to determining how procedural variables can alter dose-effect functions for cocaine self-administration is limited. Toward clarifying the role of procedural variables, responding was maintained in rats under either variable interval (VI) or fixed-ratio (FR) schedules of cocaine infusion. In addition to free-operant FR schedules, discrete-trial FR schedules were evaluated. The dose effect functions were obtained by either substituting a dose for the usual daily dose, instituting a particular dose for several sessions, or making all doses available within a session. Dose-effect functions for response rate (or number of trials with infusions for the discrete-trial FR) were bitonic for the VI and discrete-trial FR schedules but tended to be strictly decreasing for the free operant FR schedules. Responding was maintained under FR schedules by a low dose (0.083 mg/infusion) if the dose was substituted for a higher daily dose but not when made available daily. Rate of response was higher under ratio schedules at 0.17 mg/infusion when this dose occurred within the context of other higher doses within a session than when the dose was simply substituted for a higher daily dose. These data indicate that procedural variables can alter dose-response curves for cocaine self-administration. PMID- 9218278 TI - The role of dopamine in the behavioral effects of caffeine in animals and humans. AB - Dopamine has been proposed to mediate some of the behavioral effects of caffeine. This review discusses cellular mechanisms of action that could explain the role of dopamine in the behavioral effects of caffeine and summarizes the results of behavioral studies in both animals and humans that provide evidence for a role of dopamine in these effects. Caffeine is a competitive antagonist at adenosine receptors and produces a range of central and physiological effects that are opposite those of adenosine. Recently, caffeine has been shown to enhance dopaminergic activity, presumably by competitive antagonism at adenosine receptors that are colocalized and interact functionally with dopamine receptors. Thus, caffeine, as a competitive antagonist at adenosine receptors, may produce its behavioral effects by removing the negative modulatory effects of adenosine from dopamine receptors, thus stimulating dopaminergic activity. Consistent with this interpretation, preclinical behavioral studies show that caffeine produces behavioral effects similar to classic dopaminergically mediated stimulants such as cocaine and amphetamine, including increased locomotor activity, increased turning behavior in 6-hydroxydopamine-lesioned animals, stimulant-like discriminative stimulus effects, and self-administration. Furthermore, caffeine potentiates the effects of dopamine-mediated drugs on these same behaviors, and some of caffeine's effects on these behaviors can be blocked by dopamine receptor antagonists. Although more limited in scope, human studies also show that caffeine produces subjective, discriminative stimulus and reinforcing effects that have some similarities to those produced by cocaine and amphetamine. PMID- 9218279 TI - Sensitization and tolerance in psychostimulant self-administration. AB - Under some conditions, stimulant preexposure sensitizes rats to the reinforcing effects of cocaine and other stimulants, whereas under other conditions exposure decreases the reinforcing efficacy of cocaine. This paper reviews the literature on the effects of stimulant preexposure on self-administration, focusing on methodological and interpretative issues. It is concluded that both sensitization and tolerance occur following stimulant preexposure but that these two effects can be dissociated temporally, with sensitization occurring during the development of drug self-administration and tolerance occurring in response to high doses of stimulants administered to experienced self-administering rats. The relative contribution of both of these effects to compulsive drug-taking is discussed, with emphasis on the development of cocaine as a reinforcer, maintenance of self-administration, and relapse to drug-taking. PMID- 9218280 TI - Opiate and cocaine addictions: challenge for pharmacotherapies. AB - Neurobiological and behavioral studies, as well as basic and applied clinical research studies, may all contribute to the development of a pharmacotherapy for a specific addictive disease. This paper reviews recent findings from research work, primarily from one laboratory along with collaborative laboratories, that could have some relevance for the development of pharmacotherapy for cocaine dependency. The much earlier experiences of this laboratory in the development of a pharmacotherapy for opiate addiction will be addressed in the context of providing both some specific suggestions for addictive disease pharmacotherapy development and some warnings about the complexities of the introduction and implementation of a pharmacotherapy once developed. Finally, based on both the earlier perspectives and the more recent research findings, some very specific, though speculative, suggestions will be made about the development of novel pharmacotherapies for early opiate addiction, especially for cocaine abuse or addiction and prevention of relapse to cocaine use. The complex and diverse nature of the challenge for pharmacotherapy for the addictive diseases is presented, including specifically a mandate for broadening educational efforts concerning the basis of addictive diseases and the need for treatment, in parallel with the scientific efforts to develop increasingly sophisticated and targeted pharmacotherapies. PMID- 9218281 TI - Cocaine's effects on neuroendocrine systems: clinical and preclinical studies. AB - This review examines the effects of cocaine on the neuroendocrine system and summarizes findings from clinical studies of cocaine abusers and preclinical studies in rodents and rhesus monkeys. The effects of acute and chronic cocaine administration on anterior pituitary, gonadal, and adrenal hormones are described, and the functional consequences of chronic cocaine exposure are discussed. Many of cocaine's acute effects on the endocrine system are consistent with its actions as a monoamine reuptake inhibitor. Acute cocaine administration stimulates release of gonadotropins, ACTH, and cortisol or corticosterone and suppresses prolactin levels. It has been difficult to detect changes in basal levels of most hormones or alterations in hormone responsiveness to a challenge dose of cocaine or other agents after chronic cocaine treatment. Interpretation of clinical data is often complicated by polydrug abuse involving opiates and alcohol as well as cocaine. However, preclinical studies of the effects of chronic cocaine exposure on integrated neuroendocrine function have revealed disruptions of the estrous cycle in rats and the menstrual cycle in rhesus monkeys. Furthermore, the menstrual cycle disorders observed in rhesus monkeys parallel those reported in women who abuse cocaine. Much remains to be learned about cocaine's interactions with the endocrine system and the consequences of cocaine abuse for reproductive function. PMID- 9218282 TI - Mediation of the discriminative stimulus properties of cocaine by mesocorticolimbic dopamine systems. AB - This paper provides a brief review of the scientific evidence implicating the mesocorticolimbic dopamine (DA) system in modulating the discriminative stimulus properties of cocaine in rats. Briefly, systemic administration of DA releasers, reuptake inhibitors, and DA D1, D2, and putative D3 receptor agonists engendered partial to full substitution for the discriminative stimulus effects of cocaine. Dopamine D1 and D2 receptor antagonists attenuate this behavioral property of cocaine. Intracranial microinjection studies have indicated certain key limbic nuclei us loci of action for DA in mediating the discriminative stimulus effects of cocaine. Microinjections of cocaine into either DA cell body (i.e., ventral tegmental area, substantia nigra) or DA terminal regions (i.e., prefrontal cortex, central amygdala, caudate putamen) have failed to reproduce the systemic cocaine discriminative stimulus. Only infusion of cocaine into the nucleus accumbens has been demonstrated to substitute fully for the systemic effects of this psychostimulant. Interestingly, microinjections of the DA D1 receptor antagonist SCH 23390 into either the prefrontal cortex, nucleus accumbens, or central or basolateral amygdala have been demonstrated to block the discriminative stimulus properties of cocaine. Although a determination of the antagonism of the cocaine discriminative stimulus following intra-accumbens microinjection of DA D2 receptor antagonists has not been made, intra-accumbens administration of the DA D2 receptor antagonist sulpiride blocked the discriminative stimulus effects of another psychostimulant, amphetamine. 6 Hydroxydopamine lesions of DA terminals in the nucleus accumbens also attenuated the dose-effect curve for systemic administration of cocaine. Taken together, this intracranial evidence suggests that DA D1 and D2 receptors in the mesocorticolimbic system are involved in modulating the discriminative stimulus properties of psychostimulants and that the nigrostriatal DA system is not primarily involved. PMID- 9218283 TI - A global review of rabies vaccines for human use. AB - Rabies is one of the oldest known diseases of mankind, yet it has been only slightly more than 100 years since Pasteur developed the first vaccine for post exposure treatment. Since this first crude nerve tissue vaccine, numerous other rabies vaccines for human use have been developed and used with varying degrees of effectiveness and safety. When used appropriately, new cell culture vaccines provide nearly 100% protection with a high degree of safety: yet over 40,000 people world-wide die from rabies each year. Several pre- and post-exposure controlled vaccine trials and clinical studies have shown that the purified chick embryo cell (PCEC) vaccine, Rabipur, is as safe and effective as the rabies human diploid cell vaccine (HDCV), which is currently considered the gold standard. Additionally, PCEC vaccine does not result in immune-mediated hypersensitivity reactions following booster doses seen in about 6% of those receiving HDCV boosters following an initial series of HDCV. PMID- 9218284 TI - Progress and achievement of rabies control in Thailand. AB - The breakthrough in production of highly efficacious human and animal rabies vaccines has led to successful rabies control in developed countries, but rabies is still a major health problem in many developing countries. In Thailand, the new cell culture vaccines-purified chick embryo cell (PCEC), purified Vero cell (PVRV), purified duck embryo (PDEV) and human diploid cell (HDCV) are available, and since 1993 have completely replaced the nervous tissue vaccines. The impact of these cell culture vaccines has been considerable, with the number of human rabies deaths decreasing from 200-300 in the early 1980s to 74 in 1995. Rabies prevention has also focused on the vaccination of dogs, and since 1992, the year the Rabies Prevention Act was announced, every owned dog has to be vaccinated at 2-4 months of age annually thereafter. However, only about 20% of dogs have been vaccinated. In 1995, the Ministry of Agriculture and Cooperation collaborated with the Ministry of Public Health to set up a target of no human rabies deaths in 1996, and a rabies-free Thailand by the year 2000. An extensive educational campaign is underway. PMID- 9218285 TI - An ethical dilemma in rabies immunisation. AB - Rabies continues to be an important public health problem in India and many other developing countries. In India, annually some 700,000 persons are given post exposure vaccine prophylaxis using Semple (sheep brain) vaccine. It is manufactured by government institutions and given free to the public. It is presumed to be cheap, although the actual cost of production may not be low. However, it is not a safe vaccine as it causes demyelinating central or peripheral nervous system side-effects in 1/3000-7000 persons vaccinated; this adverse reaction is occasionally fatal. Cell culture rabies vaccines are also available in India; unlike the Semple vaccine they are safe and can be used for pre-exposure vaccination, but they are more expensive. The dilemma is whether it is ethically acceptable to continue to use the Semple vaccine in humans while safer products are available. What is urgently needed is a decision tree which would enable economical use of cell culture vaccines together with the backing of professional bodies in medical practice, who will declare that cost is not the only factor in choosing a rabies vaccine-safety is also of paramount importance. We must also strive to reduce the cost of cell culture vaccines. PMID- 9218286 TI - The current state of rabies prevention in Europe. AB - In Europe, from the late 1970s, the main reservoirs of rabies virus have been wild animals, mainly foxes, except in Turkey, where dog rabies account for almost all cases. Between 1977 and 1994, 198 human rabies cases were reported in Europe. In most of these cases vaccination had not been performed. In Europe, both tissue culture and nerve tissue vaccines are used with or without rabies immunoglobulin or serum, though the tissue culture vaccines have largely replaced the nerve tissue vaccines. In Hungary, where immune serum or immunoglobulin is not prescribed, no cases of human rabies among actively vaccinated individuals have been reported in the last 40 years of observation. There have, however, been two cases in which the cat, the source of infection, proved to be negative for rabies using both the fluorescent antibody and mouse inoculation tests, and therefore vaccination either did not start or was interrupted. These cases clearly question the recommendation, that a report from a reliable laboratory, indicating negative results justify cessation of treatment. PMID- 9218287 TI - Rabies and rabies research: past, present and future. AB - Rabies is probably the oldest recorded infection of mankind. The development of the first rabies vaccine by Pasteur surely had been hoped to eliminate or at least drastically reduce its incidence. However, this goal has not been achieved because rabies is maintained in many animal reservoirs, including both domestic and wild. There are still many aspects of the pathogenicity of rabies that are unknown. For example, we have no explanation for the long incubation period (up to 6 years). Furthermore, new patterns of rabies infection present a problem for epidemiologists and virologists alike. There are several cases of human rabies in which there was no history of a bite. Despite these continuing problems, there has been tremendous progress in the control of rabies. Cheap and safe vaccines for animals as well as humans have been developed. Oral vaccination of wildlife with recombinant rabies virus vaccines is beginning to reduce the incidence of rabies among foxes and raccoons. Vaccination of stray dogs could lead to the eradication of rabies in countries where dog rabies is the sole source of human exposure. PMID- 9218288 TI - The world of neurosurgery. PMID- 9218289 TI - Surgical and neurological complications in a series of 708 epilepsy surgery procedures. AB - OBJECTIVE: There are few modern data on the complications of surgery for epilepsy from the neurosurgeon's point of view. A survey of complications observed in a large current epilepsy surgery series is presented to facilitate the assessment of a risk:benefit ratio, which must be known when planning for epilepsy surgery and counseling patients. METHODS: A series of 429 consecutive patients operated on during 6.5 years in the newly established University of Bonn epilepsy surgery program was, in part, retrospectively, and, in larger part, prospectively analyzed for complications originating from 279 invasive diagnostic procedures and 429 therapeutic procedures. Neuropsychological and psychiatric complications as well as the rate of failure to control seizures are not addressed in this article. RESULTS: Two hundred and seventy-nine temporal operations, 59 frontal operations, 22 other extratemporal operations, 33 callosotomies, 3 multilobectomies, and 33 hemispherectomies were performed. Complications were grouped into general surgical and neurological complications. No mortality resulted from 708 invasive procedures. Two hundred and seventy-nine invasive diagnostic procedures (various combinations of strip, grid, and depth electrode insertions) resulted in 3.6% transient morbidity (2.9% surgical complications, 0.7% neurological complications) and 0.7% permanent morbidity (dysphasia). During 429 therapeutic procedures, 33 surgical complications were encountered. None of these resulted in permanent morbidity, except for the necessity for permanent shunt insertion in three patients. Wound infection was the most frequent surgical complication, but we were able to demonstrate a steady decrease during the 6.5 year observation period. The total rate of neurological complications in 429 therapeutic procedures was 5.4%, with 3.03% causing transient morbidity and 2.33% causing permanent morbidity. CONCLUSION: Our data indicate that epilepsy surgery can be performed with an acceptable rate of resultant morbidity. The indications for epilepsy surgery, the learning curve determined, and the results from other series are discussed in the light of these figures. PMID- 9218290 TI - Continuous assessment of the cerebral vasomotor reactivity in head injury. AB - OBJECTIVE: Cerebrovascular vasomotor reactivity reflects changes in smooth muscle tone in the arterial wall in response to changes in transmural pressure or the concentration of carbon dioxide in blood. We investigated whether slow waves in arterial blood pressure (ABP) and intracranial pressure (ICP) may be used to derive an index that reflects the reactivity of vessels to changes in ABP. METHODS: A method for the continuous monitoring of the association between slow spontaneous waves in ICP and arterial pressure was adopted in a group of 82 patients with head injuries. ABP, ICP, and transcranial doppler blood flow velocity in the middle cerebral artery was recorded daily (20- to 120-min time periods). A Pressure-Reactivity Index (PRx) was calculated as a moving correlation coefficient between 40 consecutive samples of values for ICP and ABP averaged for a period of 5 seconds. A moving correlation coefficient (Mean Index) between spontaneous fluctuations of mean flow velocity and cerebral perfusion pressure, which was previously reported to describe cerebral blood flow autoregulation, was also calculated. RESULTS: A positive PRx correlated with high ICP (r = 0.366; P < 0.001), low admission Glasgow Coma Scale score (r = 0.29; P < 0.01), and poor outcome at 6 months after injury (r = 0.48; P < 0.00001). During the first 2 days after injury, PRx was positive (P < 0.05), although only in patients with unfavorable outcomes. The correlation between PRx and Mean index (r = 0.63) was highly significant (P < 0.000001). CONCLUSION: Computer analysis of slow waves in ABP and ICP is able to provide a continuous index of cerebrovascular reactivity to changes in arterial pressure, which is of prognostic significance. PMID- 9218291 TI - Direct intraoperative measurement of human brain temperature. AB - OBJECTIVE: Because hypothermia enhances human tolerance for cerebral ischemia, profound hypothermia is induced in many centers so that the circulation can be arrested while clips are applied to high-risk giant cerebral aneurysms. Brain temperature is measured directly with an intracerebral probe that avoids the uncertainty of surrogate monitoring. However, when there is a large thermal gradient between brain temperature and that of the operating room, even direct measurements can sometimes be misleading. This study was undertaken to determine how deeply a thermal sensor must be embedded in the cerebral parenchyma to ensure that the ambient environment does not distort the measurement of brain temperature. METHODS: Each of 39 normothermic patients had a thermocouple sensor inserted into a temporal lobe seizure focus just before its resection. Brain temperature was measured as the sensor was withdrawn in stages. RESULTS: At both 3 and 2 cm beneath the cortical surface, the temperature of the brain was essentially the same. However, when the sensor was withdrawn to 1 cm, recorded temperature decreased from 35.7 +/- 0.9 to 34.3 +/- 1.4 degrees C (P < 0.001) and irrigation in the vicinity caused major thermal change. At shallower depths, even lower brain temperatures were recorded. No morbidity was attributable to the temperature measurements. CONCLUSION: Direct intraoperative measurement of human brain temperature is feasible and safe, but accuracy requires that the temperature sensor be inserted at least 2 cm into the cerebral cortex. PMID- 9218292 TI - Neurological and psychosocial outcome 4 to 7 years after subarachnoid hemorrhage. AB - OBJECTIVE: Previous studies have demonstrated that many patients with good neurological outcomes still experience excessive fatigue, cognitive impairments, and lowered work status 1 year after subarachnoid hemorrhage (SAH). Does recovery continue for many years or are survivors of SAH left with permanent disabilities? We describe the long-term outcome. METHODS: Approximately 50% (n = 123) of the patients who survived SAH for more than 3 years from a population of 1.5 million and who had participated in research studies at the time of their SAH were interviewed 4 to 7 years later by telephone or questionnaire. Participants did not differ from the 126 unsurveyed survivors in age, gender, SAH grades, aneurysm sites, or complications. RESULTS: Most patients thought their recovery to be satisfactory to good, although some reported memory problems (41%), headaches (16.5%), daytime sleepiness (35%), problems sleeping at night (26%), a reduced ability to work (20%), and a changed personality (48.3%). Many had reduced their smoking and drinking. Each of 24 of the 121 participants for whom seizure data were available (all with clipped aneurysms) had suffered at least one seizure, but only each of 10 had suffered two or more seizures since hospital discharge. Relative youth was the only significant predictor of seizures, with strong trends observed between seizures and a poor Glasgow Outcome Scale score at 10 weeks or between seizures and an ischemic neurological deficit. No evidence for the effectiveness of prophylactic anticonvulsants was demonstrated. CONCLUSION: Survivors of SAH continue to recover for years and develop good coping skills and a positive attitude toward their recovery, even when they experience ongoing problems. Few are left with disabling headaches or epilepsy. PMID- 9218293 TI - Hemodynamic instability after carotid endarterectomy: risk factors and associations with operative complications. AB - OBJECTIVE: To examine the incidences of hypertension, hypotension, and bradycardia after carotid endarterectomy (CEA) and to identify any hemodynamic variables predictive of postoperative stroke, death, or cardiac complications. METHODS: Retrospective population-based cohort study of 291 consecutive patients undergoing CEA using hospital chart review. Hemodynamic data collected from time of arrival in the recovery room until the end of the 1st postoperative day. Primary and secondary outcome events were stroke or death within 30 days of surgery and any postoperative cardiac complication (angina, congestive heart failure, dysrhythmia, or myocardial infarction), respectively. RESULTS: The incidences of postoperative hypertension (systolic blood pressure > 220 mm Hg), hypotension (systolic blood pressure < 90 mm Hg), and bradycardia (pulse < 60 beats/min) were 9% (26 of 290 cases), 12% (36 of 290 cases), and 55% (159 of 290 cases), respectively. The stroke or death rate was 5.2% (15 of 291 cases). Postoperative hypertension was associated significantly with stroke or death (P = 0.04) and by a statistical trend with cardiac complications (P = 0.07). Independent preoperative risk factors for postoperative hypertension by multivariate analysis included angiographic intracranial carotid stenosis greater than 50%, cardiac dysrhythmia, preoperative systolic blood pressure greater than 160 mm Hg, neurological instability, and renal insufficiency. Postoperative hypotension and bradycardia did not correlate with primary or secondary outcomes. CONCLUSION: Hemodynamic instability was commonly observed after CEA, but only postoperative hypertension was associated with stroke or death and, possibly, with cardiac complications. Patients undergoing CEA, especially those at risk for postoperative hypertension, may be monitored best in settings suited to the expeditious management of neurological and cardiovascular emergencies. PMID- 9218294 TI - Interstitial chemotherapy with carmustine-loaded polymers for high-grade gliomas: a randomized double-blind study. AB - OBJECTIVE: To find out the effect of carmustine (bischloroethyl-nitrosourea) combined with a biodegradable polymer in the treatment of malignant (Grades III and IV) gliomas, applied locally, at the time of the primary operation. METHODS: Prospective, randomized double-blind study of an active treatment group versus a placebo group. Conducted at the Departments of Neurosurgery of the University Hospitals of Helsinki, Tampere, and Turku in Finland and Trondheim in Norway. The study consisted of 32 patients (16 in each treatment group) enrolled between March 23, 1992, and March 19, 1993. The study was planned to include 100 patients but had to be terminated prematurely, because the drug that was being used had become unobtainable. The main outcome measures included the survival times of patients after the operations and the application of an active drug or placebo. RESULTS: The median time from surgery to death was 58.1 weeks for the active treatment group versus 39.9 weeks for the placebo group (P = 0.012). For 27 patients with Grade IV tumors, the corresponding times were 39.9 weeks for the placebo group and 53.3 weeks for the active treatment group (P = 0.008). At the end of the study, six patients were still alive, five of whom belonged to the active treatment group. CONCLUSION: Carmustine applied locally in a biodegradable polymer at the time of primary operation, seems to have a favorable effect on the life span of patients with high-grade gliomas. PMID- 9218296 TI - Cerebellar abscess: the significance of cerebrospinal fluid diversion. AB - OBJECTIVE: Cerebellar abscesses that are often ominously silent have a significant mortality. Sudden total occlusion of cerebrospinal fluid (CSF) pathways makes an aggressive surgical approach mandatory. Our neurosurgical unit at Wentworth Hospital, Durban, South Africa, prospectively instituted a protocol for patients with cerebellar abscesses with reference to CSF diversion with the aim of improving outcome. Our 13-year experience with this approach to cerebellar abscesses is presented. METHODS: Since 1983, patients with cerebellar abscesses have been managed according to a standard protocol. In 1987, a policy of aggressive CSF diversion was prospectively instituted. This involved immediate CSF diversion in any patient with over or incipient hydrocephalus, even if fully conscious. The associated hydrocephalus was diagnosed on initial computed tomographic scans. CSF diversion was performed by means of a ventricular drain, inserted in the reception area under local anesthesia. The period from January 1983 to December 1995 was analyzed, and the impact of aggressive CSF diversion on patient outcome was evaluated. RESULTS: Seventy-seven patients with cerebellar abscesses during the 13-year period under review were studied. Thirty-four patients were treated before the introduction of the policy of aggressive CSF diversion. Of these patients, 10 died, resulting in a mortality of 29% and a morbidity of 21%. Forty-three patients were treated after the institution of the new policy of CSF diversion. Of these patients, five died, resulting in a mortality rate of 11.6% and a morbidity rate of 14%. CONCLUSION: Although surgical drainage of a cerebellar abscess and eradication of the primary septic source and appropriate antibiotic coverage are necessary, the management of hydrocephalus, or even incipient hydrocephalus, is of paramount importance. PMID- 9218295 TI - Acoustic neuromas: results of current surgical management. AB - OBJECTIVE: In this article, we review the surgical outcomes of 179 patients with acoustic neuromas. METHODS: Most of the tumors (84%) were operated on using a retrosigmoid, transmeatal approach. A transpetrosal, retrosigmoid approach was used in 10% of the patients, most of whom had large tumors. The translabyrinthine (4%) and transmastoid, transpetrosal, partial labyrinthectomy approaches (2%) were used selectively. The operative approaches are discussed. Tumors were categorized according to their cerebellopontine angle dimensions as small (< 2 cm), medium (2.0-3.9 cm), and large (> or = 4 cm). RESULTS: House-Brackmann evaluation of postoperative facial nerve function revealed excellent results (Grade I or II) in 96% of small tumors, 74% of medium tumors, and 38% of large tumors. A fair postoperative function (Grade III or IV) was achieved in 4% of small tumors, 26% of medium tumors, and 58% of large tumors. Functional hearing preservation, defined as Gardner-Robertson Class I or II, was achieved in 48% of small tumors and 25% of medium tumors. Hearing was not preserved in any of the three patients with large tumors in whom hearing preservation was attempted. Treatment complications consisted mainly of cerebrospinal fluid leakage (15% of the patients). The majority of the patients who experienced cerebrospinal fluid leakage were treated successfully with lumbar spinal drainage; only four patients (2% of the total group) required subsequent surgery for correction of cerebrospinal fluid leakage. There were two deaths (1%) in this series. One death occurred as the result of myocardial infarction and the other as the result of severe obstructive lung disease. One patient sustained disability because of cerebellar and brain stem injury. Complete tumor resection was accomplished in 99% of the patients, and there was no evidence of recurrence in this group. Only 1 of the 179 patients underwent incomplete tumor resection; he required subsequent surgery for symptomatic tumor regrowth. Our patient follow-up had a mean duration of 70 months and a median of 65 months (range, 3-171 mo). CONCLUSION: Our results are similar to those of other large microsurgical series of acoustic neuromas. Unless a patient has major medical problems, microsurgery by an experienced team of surgeons is preferred over radiosurgery. PMID- 9218297 TI - Idiopathic spinal epidural lipomatosis. AB - OBJECTIVE: Spinal epidural lipomatosis (SEDL) is a rare disorder often associated with the administration of exogenous steroids or the elevation of endogenous steroids. Spinal epidural lipomatosis develops in some patients in the absence of elevated steroid levels. The limited information known about idiopathic SEDL comes predominantly from isolated case reports. We proposed to study our experience with idiopathic SEDL and to review the literature. METHODS: We identified eight symptomatic patients with idiopathic SEDL treated at our institution, which is the largest series reported. All patients were male and obese by body mass index (> 27.5 kg/m2). The mean age of the patients was 35.4 years. Idiopathic SEDL was equally distributed between the thoracic and lumbar spine. Six patients underwent laminectomy and fat debulking with good postoperative results; two patients were treated with a weight loss diet, which resulted in the relief of symptoms after losing > 15 kg each. RESULTS AND CONCLUSION: A review of our patients in conjunction with other reported cases reveals the following: 1) idiopathic SEDL occurs almost exclusively in the obese population; 2) idiopathic SEDL seems to occur with equal frequency between the thoracic and lumber spine; 3) a strong male predominance exists; 4) thoracic SEDL presents at an earlier age compared with lumbar SEDL; 5) surgical decompression remains the treatment of choice for the immediate relief of symptoms. Our experience suggests that idiopathic epidural lipomatosis may be a pathological entity that has been underdiagnosed. PMID- 9218298 TI - Median nerve compression can be detected by magnetic resonance imaging of the carpal tunnel. AB - OBJECTIVE: Clinically symptomatic carpal tunnel syndrome is not necessarily accompanied by impaired nerve conduction values. Surgical decompression, however, may immediately lead to complete and lasting relief of symptoms in these patients. Because minimally invasive techniques have reduced perioperative morbidity and vocational impairment related to operative decompression, the decision to decompress symptomatic patients (despite still unimpaired nerve conduction values) might be subject to discussion in the future. New diagnostic tools may be helpful in deciding which therapeutical options to choose. When the wrist is held either in flexion or in extension, the carpal tunnel pressure increases. To investigate the dynamic changes of the carpal tunnel shape during wrist motion, as well as the variations of space for the median nerve and its signal intensity in T2-weighting, magnetic resonance imaging (MRI) was performed on patients and healthy volunteers alike. Restitution and the persistence of pathological findings were assessed pre- and postoperatively. METHODS: MRI (1.0 T) was performed on 20 wrists of patients with clinical symptoms of carpal tunnel syndrome (CTS) and pathological nerve conduction values. Healthy volunteers (20 wrists) were matched according to sex and age. MRI was performed in neutral, 45 degree extension, and 45-degree wrist flexion positions. T2-weighted signal intensity of the median nerve was measured in 18 patients pre- and postoperatively. RESULTS: The cross-sectional area of the carpal tunnel in patients with CTS tends to be smaller than that found in nonsymptomatic volunteers. The cross-sectional area of the carpal tunnel decreases during wrist flexion at the pisiform and hamate level. During wrist extension, the cross sectional area of the carpal tunnel decreases at the level of the pisiform. During extension, it increases at the level of the hamate. The cross-sectional area of the median nerve showed an increase at the pisiform level (P < 0.05), a flattening of the median nerve at the hamate hook level (P < 0.05), and palmar deviation of the flexor retinaculum at the pisiform and hamate hook level (P < 0.001). This was significantly greater in CTS patients than in individuals with normal wrists. Postoperatively, the distal flattening of the median nerve recovered in 94% of the cases reviewed. Although the signal intensity of the median nerve on T2-weighted images decreased by 67%, the motor latency recovered in only 39% of the cases. CONCLUSION: The carpal tunnel was smaller in CTS patients than in healthy volunteers. During flexion and extension, the space available for the median nerve narrows. This may lead to potential median nerve compression. MRI is accurate and reliable for diagnosis and postoperative follow up of carpal tunnel syndrome. In cases with obvious clinical symptoms and yet not measurably impaired median nerve conduction values, it may be helpful in making a decision for surgical decompression. PMID- 9218299 TI - Decompressive bifrontal craniectomy in the treatment of severe refractory posttraumatic cerebral edema. AB - OBJECTIVE: The management of malignant posttraumatic cerebral edema remains a frustrating endeavor for the neurosurgeon and the intensivist. Mortality and morbidity rates remain high despite refinements in medical and pharmacological means of controlling elevated intracranial pressure; therefore, a comparison of medical management versus decompressive craniectomy in the management of malignant posttraumatic cerebral edema was undertaken. METHODS: At the University of Virginia Health Sciences Center, 35 bifrontal decompressive craniectomies were performed on patients suffering from malignant posttraumatic cerebral edema. A control population was formed of patients whose data was accrued in the Traumatic Coma Data Bank. Patients who had undergone surgery were matched with one to four control patients based on sex, age, preoperative Glasgow Coma Scale scores, and maximum preoperative intracranial pressure (ICP). RESULTS: The overall rate of good recovery and moderate disability for the patients who underwent craniectomies was 37% (13 of 35 patients), whereas the mortality rate was 23% (8 of 35 patients). Pediatric patients had a higher rate of favorable outcome (44%, 8 of 18 patients) than did adult patients. Postoperative ICP was lower than preoperative ICP in patients who underwent decompression (P = 0.0003). Postoperative ICP was lower in patients who underwent surgery than late measurements of ICP in the matched control population. A statistically significant increased rate of favorable outcomes was seen in the patients who underwent surgery compared to the matched control patients (15.4%) (P = 0.014). All patients who exhibited sustained ICP values above 40 torr and those who underwent surgery more than 48 hours after the time of injury did poorly. Evaluation of the 20 patients who did not fit into either of those categories revealed a 60% rate of favorable outcome and a statistical advantage over control patients (P = 0.0001). CONCLUSION: Decompressive bifrontal craniectomy provides a statistical advantage over medical treatment of intractable posttraumatic cerebral hypertension and should be considered in the management of malignant posttraumatic cerebral swelling. If the operation can be accomplished before the ICP value exceeds 40 torr for a sustained period and within 48 hours of the time of injury, the potential to influence outcome is greatest. PMID- 9218300 TI - Cine magnetic resonance imaging of spinal intradural arachnoid cysts. AB - OBJECTIVE: The diagnostic value of cine magnetic resonance imaging (CMRI) that visualizes fluid and tissue movement was evaluated in patients with spinal intradural arachnoid cysts, a rare but increasingly detected cause of spinal cord dysfunction. METHODS: Four patients with thoracic spinal intradural arachnoid cysts were investigated with conventional T1- and T2-weighted and cardiac-gated CMRI. Four normal volunteers also underwent CMRI for comparison. RESULTS: Sagittal T1- and T2-weighted imaging showed lesions as an abnormal widening of the posterior spinal subarachnoid space, but mixed high- and low-signal intensities on T2-weighted imaging suggested cystic lesions. CMRI, using 16 to 20 sagittal gradient echo images during the cardiac cycle of normal volunteers, indicated synchronous signal changes along the subarachnoid space, suggesting a smooth cerebrospinal fluid flow. CMRI of patients detected that the caudal or cranial direction of the high-signal propagation suddenly reversed at some locations (as if rebounding) in an asynchronous fashion along the lesion (asynchronous rebound phenomenon), which was well demonstrated by the closed-loop video mode. Cystectomy revealed that the cysts consisted of multiple lobules and that the asynchronous rebound phenomenon corresponded with some boundaries of cyst lobules. CMRI also visualized dynamic spinal cord compression by the cyst. CONCLUSION: CMRI can demonstrate abnormal fluid flow and spinal cord compression caused by a spinal intradural arachnoid cyst. PMID- 9218301 TI - Thrombotic, infectious, and procedural complications of the jugular bulb catheter in the intensive care unit. AB - OBJECTIVE: An assessment of the thrombotic, infectious, and technical complications of continuous jugular bulb catheter monitoring in the intensive care unit (ICU) was made. METHODS: Over a 1-year period, 44 patients suffering from traumatic brain injury, subarachnoid hemorrhage, or stroke received jugular bulb catheter monitoring in the ICU. They were followed for catheter insertion complications and the development of bacteremia. In 20 patients chosen randomly, an ultrasonographic evaluation was performed after removal of the catheter for an assessment of internal jugular vein thrombosis. RESULTS: Of the 44 patients, 1 became bacteremic; the source was identified as a thoracostomy site. Among the complications related to the 44 catheter insertions, there were 2 instances of carotid artery puncture (4.5%), 1 misplaced catheter (thoracic placement), and 1 clinically insignificant hematoma. Of the 20 patients investigated with ultrasonography, 8 (40%) had nonobstructive, subclinical internal jugular vein thrombi after jugular bulb catheter monitoring (95% confidence interval, 19-61%). The median monitoring duration was 3 days (range, 1-6 d). No clinical factor was identified to be associated with thrombus formation. CONCLUSION: We conclude the following: 1) the risk of bacteremia related to the jugular bulb catheter was negligible; 2) complications related to catheter insertion were rare and clinically insignificant; and 3) the incidence of subclinical internal jugular vein thrombosis after jugular bulb catheter monitoring is considerable. Although it is worthy to note this complication, no patient with a thrombus became symptomatic in the present series. The risk-benefit assessment of this monitoring technique must include consideration of subclinical thrombosis. PMID- 9218302 TI - Complications in patients with palmar hyperhidrosis treated with transthoracic endoscopic sympathectomy. AB - OBJECTIVE: To assess the complications in a group of patients with palmar hyperhidrosis treated with transthoracic endoscopic sympathectomy. The extraordinarily high incidence of postoperative compensatory hyperhidrosis in our series is stressed and explained. METHODS: The retrospective study included chart reviews and outpatient assessments. Seventy-two patients underwent T2 or T2-T3 endoscopic sympathectomy for primary palmar hyperhidrosis. Patients' hyperhidrosis severity, precipitating factors, postoperative complications, surgical results, and satisfaction were assessed. Severity of palmar hyperhidrosis and compensatory hyperhidrosis was classified by two grading scales. RESULTS: The success rate of sympathectomy was 93%. All patients except one suffered from compensatory sweating, which was the main cause of patients' dissatisfaction postoperatively. Seventeen percent of the patients (12 of 72 patients) experienced new symptoms of gustatory sweating (facial sweating associated with eating). Twenty-one patients experienced other complications, including pneumothorax, Horner's syndrome, nasal obstruction, and intercostal neuralgia. CONCLUSION: Transthoracic endoscopic sympathectomy is an effective and simple modality to treat palmar hyperhidrosis. However, all patients need to be warned of the common complications, particularly compensatory hyperhidrosis, before surgery. PMID- 9218303 TI - Smoking and the human vertebral column: a review of the impact of cigarette use on vertebral bone metabolism and spinal fusion. AB - Chronic cigarette consumption has significant adverse effects on the human spinal column. Multiple mechanisms induced by tobacco use lead to less strong, less healthy, mineral-deficient vertebrae with reduced bone blood supply and fewer and less functional bone-forming cells among chronic smokers. Compared to nonsmokers, chronic smokers develop advanced bony degradation, are more likely to suffer from spinal column degenerative disease, and seem more susceptible to traumatic vertebral injury. Spinal fusion procedures in chronic smokers are less often clinically and radiographically successful, compared to similar procedures performed among nonsmokers for definitive biological, physiological, and mechanical reasons. PMID- 9218304 TI - Three-dimensional computed tomographic angiography in detection of cerebral aneurysms in acute subarachnoid hemorrhage. AB - OBJECTIVE: Three-dimensional computed tomographic angiography (CTA) is a recently developed imaging modality. We demonstrate the value of this noninvasive method in replacing digital subtraction angiography (DSA) in the detection of aneurysms of the circle of Willis in patients with subarachnoid hemorrhage admitted to our institution. METHODS: A helical acquisition was performed for computed tomographic scans obtained for 120 patients with a 1 mm per second table speed and a 1-mm collimation, 1:1 pitch. Axial source images were transferred on a console Advantage Windows workstation (General Electric, Milwaukee, WI) and CTA was obtained using maximum intensity projection reconstruction. All patients had undergone DSA of the circle of Willis (80 patients preoperatively and 40 postoperatively). RESULTS: A total of 129 aneurysms were detected in 107 patients. Three-dimensional CTA disclosed nothing abnormal in 13 patients. Ninety two patients sustained one aneurysm, 10 patients sustained two, 3 patients sustained three, and 2 patients sustained four. All results were confirmed by DSA. In two cases, aneurysms of the middle cerebral artery were defected by CTA but not by DSA. When using angiographic views, the aneurysm was always masked by a branch of the middle cerebral artery. CONCLUSION: The sensitivity of three dimensional CTA is comparable with that of DSA, and its specificity is 100%. Because CTA is simple, quick, noninvasive, and reliable, we think that it can eventually replace DSA. PMID- 9218305 TI - Stereotactic localization with magnetic resonance imaging: a phantom study to compare the accuracy obtained using two-dimensional and three-dimensional data acquisitions. AB - OBJECTIVE: To compare the accuracy of stereotactic localization using magnetic resonance imaging (Siemens 1.5-T Magnetom; Siemens, Erlangen, Germany) with two dimensional and three-dimensional data acquisition techniques. METHODS: A phantom study was performed in which the coordinates of an array of rods were determined from images in both two-dimensional and three-dimensional studies and compared with measured values in a series of transverse, coronal, and sagittal images. RESULTS: The results demonstrated a distinct advantage in using three-dimensional acquisition; an error greater than 2 mm was identified in only 0.8% of the imaged volume, compared with 12% of the imaged volume in the two-dimensional study. CONCLUSION: The results indicated that more accurate stereotactic localization is achieved with a three-dimensional acquisition. PMID- 9218306 TI - A new subarachnoid hemorrhage grading system based on the Glasgow Coma Scale: a comparison with the Hunt and Hess and World Federation of Neurological Surgeons Scales in a clinical series. AB - OBJECTIVE: Although the Hunt and Hess Scale (HHS) and World Federation of Neurological Surgeons Scale (WFNSS) are the most widely used subarachnoid hemorrhage (SAH) grading systems, neither system has achieved universal acceptance. We propose a simplified grading system based entirely on the Glasgow Coma Scale (GCS), which compresses the 15-point GCS into five grades that are comparable with those of the HHS and WFNSS. We refer to this system as the GCS grading system and present a direct comparison with the HHS and WFNSS for predictive value regarding patient outcome and interrater reliability. METHODS: We reviewed 291 consecutive patients with aneurysms treated at our institution between January 1992 and January 1996 and compared the admission grades from the GCS, WFNSS, and HHS with outcome measures at discharge from hospitalization. The Glasgow Outcome score was used as the major outcome measure to evaluate the predictive value of the three scales. Mortality and length of stay (LOS) were also evaluated as outcome measures. The predictive value of each scale was tested with an ordinal logistic regression model for Glasgow Outcome score, a logistic regression model for mortality data, and a linear regression model for LOS. RESULTS: Using the logistic regression model, the GCS was the best predictor of discharge Glasgow Outcome score, with an odds ratio of 2.585 (P = 0.0001), compared with 2.311 (P = 0.0001) for the WFNSS and 2.262 (P = 0.0001) for the HHS. Using mortality data in the logistic model, the HHS was the best predictor, with an odds ratio of 3.391 (P = 0.0001), compared with 2.859 (P = 0.0001) for the GCS and 2.560 (P = 0.0001) for the WFNSS. Each of the three scales had a high predictive value for LOS, using a linear model. We discuss, however, the problematic nature of LOS as an outcome measure for SAH. Interrater reliability for each scale was evaluated using kappa statistics, based on 15 additional patients evaluated prospectively, and showed that the GCS grade also had the greatest interrater reliability, with a kappa of 0.46 (P = 0.0002), compared with 0.41 (P = 0.0005) for the HHS and 0.27 (P = 0.027) for the WFNSS. CONCLUSION: We conclude that the GCS grade has equal or greater predictive value regarding outcome after SAH than do the currently used grading systems and that it has greater reproducibility across observers. Broader familiarity with the GCS among medical and paramedical personnel may further enhance the usefulness of the GCS grade over the HHS and WFNSS in providing a standardized, universally accepted grading system for SAH. PMID- 9218307 TI - The jugular foramen: microsurgical anatomy and operative approaches. AB - The jugular foramen, based on these studies of microsurgical anatomy, is divided into three compartments: two venous and a neural or intrajugular compartment. The venous compartments consist of a larger posterolateral venous channel, the sigmoid part, which receives the flow of the sigmoid sinus, and a smaller anteromedial venous channel, the petrosal part, which receives the drainage of the inferior petrosal sinus. The petrosal part forms a characteristic venous confluens by also receiving tributaries from the hypoglossal canal, petroclival fissure, and vertebral venous plexus. The petrosal part empties into the sigmoid part through an opening in the medial wall of the jugular bulb between the glossopharyngeal nerve anteriorly and the vagus and accessory nerves posteriorly. The intrajugular or neural part, through which the glossopharyngeal, vagus, and accessory nerves course, is located between the sigmoid and petrosal parts at the site of the intrajugular processes of the temporal and occipital bones, which are joined by a fibrous or osseous bridge. The glossopharyngeal, vagus, and accessory nerves penetrate the dura on the medial margin of the intrajugular process of the temporal bone to reach the medial wall of the internal jugular vein. The operative approaches, which access the foramen and adjacent areas and are demonstrated in a stepwise manner, are the postauricular transtemporal, retrosigmoid, extreme lateral transcondylar, and preauricular subtemporal infratemporal approaches. PMID- 9218308 TI - Radioprotective effects of the 21-aminosteroid U-74389G for stereotactic radiosurgery. AB - OBJECTIVE: Future improvements in the results of stereotactic radiosurgery will be related to better patient selection, dose planning, radiosensitization of the target, and, possibly, protection of the brain surrounding the target. 21 Aminosteroids may provide protection against brain radiation injury by inhibition of lipid peroxidation and a selective action on vascular endothelium. We hypothesized that the 21-aminosteroid U-74389G would reduce radiosurgery-related brain injury without attenuating the target volume response. METHODS: One hundred and forty-five rats were divided into four experimental groups before undergoing radiosurgery: control (n = 47); low-dose U-74389G (5 mg/kg of body weight, n = 30); high-dose U-74389G (15 mg/kg, n = 20); and methylprednisolone (2 mg/kg, n = 48). The drug was administered 1 hour before radiosurgery (4-mm gamma knife collimator) of the normal rat frontal lobe (single-fraction maximum doses of 50, 100, or 150 Gy) was performed. All brains underwent histological examination at 90 or 150 days to evaluate the diameters of necrosis and the findings of radiation-induced vasculopathy, brain edema, and gliosis. RESULTS: None of the animals that received 50-Gy radiation developed histological changes, whereas all of the animals that received 150-Gy radiation developed radiation necrosis without drug-induced protection from vascular changes or edema. In animals receiving 100-Gy radiation, high-dose aminosteroid reduced radiation-induced vasculopathy at 90 days (P = 0.06) and at 150 days (P = 0.02) and prevented regional edema at 90 days (P = 0.01) and at 150 days (P = 0.03). Low-dose aminosteroid and corticosteroid provided no protection. CONCLUSION: The 21 aminosteroid U-74389G provided protection after a single intravenously administered dose of 15 mg/kg against radiation-induced vasculopathy and edema. High-dose 21-aminosteroids seem to have optimal properties for radiosurgery, surrounding brain protection without reducing the therapeutic effect desired within the target volume. PMID- 9218309 TI - The protective effect of dexamethasone against radiation damage induced by interstitial irradiation in normal monkey brain. AB - OBJECTIVE: The protective effect of dexamethasone against radiation damage is unclear. We examined the effect of early treatment of high-dose dexamethasone on iridium-192-induced damage to normal brain tissue. METHODS: Brain damage induced by interstitial irradiation with iridium-192 was evaluated with sequential magnetic resonance imaging and proton magnetic resonance spectroscopy in 11 adult monkeys, with or without short-term high-dose dexamethasone treatment. Dexamethasone (1 mg/kg of body weight/d) was administered intramuscularly to five irradiated animals every 24 hours, beginning 2 days before and ending 7 days after irradiation. Magnetic resonance imaging and proton magnetic resonance spectroscopy were performed 1 week, 1 month, and 3 months after irradiation. RESULTS: Magnetic resonance imaging performed 1 week after irradiation revealed marked edema in five nontreated animals. In dexamethasone-treated animals, the volume of edema was reduced significantly, compared to that of nontreated animals, 1 week and 1 month after irradiation. The volume of ring enhancement in dexamethasone-treated animals was also reduced significantly, compared to that of nontreated animals, 3 months after the irradiation. Proton magnetic resonance spectroscopy spectra revealed that N-acetylaspartate and choline peaks were reduced 1 week after irradiation in both groups. However, there were no statistically significant differences between the two groups at any time points. CONCLUSION: These results suggest that dexamethasone treatment may have an antiedema effect at an early stage and may modify subsequent development of vascular and inflammatory changes but may have no effect of preventing radiation induced necrosis and the reduction of N-acetylaspartate after brachytherapy. PMID- 9218310 TI - Rapid saccular aneurysm induction by elastase application in vitro. AB - OBJECTIVE: To develop a new saccular aneurysm model in vitro using elastase to study aneurysm initiation, growth, and rupture and to create a new in vivo aneurysm model to test endovascular therapies. METHODS: Seventeen common carotid arteries excised from freshly killed pigs and sheep were treated with seven different methods of elastase delivery. The arteries were mounted in a saline filled flow chamber. They received pulsatile flow for 48 hours, or until the resulting aneurysms ruptured. Changes were continuously monitored with video camera recordings and validated with histological sections. RESULTS: All eight arteries treated topically, either on the intimal or on the adventitial surface, with elastase concentrations greater than 1 U/mm2, developed saccular aneurysms; five of them ruptured within 48 hours. All four arteries treated with surface concentrations of 0.1 U/mm2 via microcatheter infusion into the lumen developed fusiform aneurysms. None of the arteries that received surface concentrations less than 0.1 U/mm2 developed aneurysms. Histological sections revealed a reduced number of cellular element in a stretched collagen matrix at the dome of the saccular aneurysms. CONCLUSION: After empirically testing several methods of elastase delivery, we were able to induce saccular, bifurcation-type aneurysms in animal arterial specimens. These aneurysms are histologically similar and more authentic than surgical models. The procedure is easy and reproducible. Our results suggest a possible enzymatic role in aneurysm formation and highlight the dramatic effects of selective arterial elastic damage. Also, the rapid growth of our experimental aneurysms may reflect the speed of the natural process. PMID- 9218311 TI - Morphological changes of intraparenchymal arterioles after experimental subarachnoid hemorrhage in dogs. AB - OBJECTIVE: Morphological and microcirculatory changes in intraparenchymal vessels after subarachnoid hemorrhage (SAH) have not yet been fully clarified. We conducted this experimental study to investigate the serial morphological changes of intraparenchymal arterioles after SAH. METHODS: SAH was produced by injecting autologous arterial blood into the cisterna magna twice at 48-hour intervals in 30 dogs. The dogs were killed 3, 7, or 14 days after SAH, and then perfusion fixed specimens of both anterior sylvian giri were obtained by using two methods. Microvascular corrosion casts produced by arterial injection of polyester resin were examined using scanning electron microscopy, and the widths of 40 arterioles of each animal were measured. Sectioned slices from the brain surface to 500 microns deep were examined by light microscopy, and external diameter, internal diameters, and wall thickness of the arterioles at depths of 50, 200, and 500 microns from the brain surface were morphometrically evaluated in 40 arterioles of each animal. In control animals receiving cisternal injections of mock cerebrospinal fluid (n = 10) and in healthy control animals (n = 10), the same examination and evaluation were performed. RESULTS: Corrosion casts of arterioles showed tapered narrowing with folding after SAH, and the width of the arterioles significantly decreased 3 and 7 days after SAH (P < 0.01). Morphometric examination by light microscopy showed a significant decrease of internal diameter of arterioles associated with a significant increase of wall thickness at any depth from the brain surface 3 and 7 days after SAH (P < 0.05 or P < 0.01). These findings improved 14 days after SAH. Control animals receiving cisternal injections of mock cerebrospinal fluid showed no significant differences compared with healthy control animals. CONCLUSION: These results suggest that constriction of intraparenchymal arterioles occurs after SAH and may contribute to delayed cerebral ischemia. PMID- 9218312 TI - Tumor blood flow and the cytotoxic effects of estramustine and its constituents in a rat glioma model. AB - OBJECTIVE: Estramustine (EaM) is a conjugate of nor-nitrogen mustard (NNM) and 17 beta-estradiol (E2) that has cytotoxic and radiosensitizing effects on experimental malignant glioma. Its mechanism of action is only partly understood. To further investigate the mechanism in vivo, the effects on tumor blood flow (TBF) and tumor growth were analyzed. METHODS: TBF was measured by radioactive microspheres, and tumor growth was measured by weight. Apoptosis was evaluated by in situ end labeling and gel electrophoresis. The effects of the constituents NNM and E2 were also evaluated. RESULTS: EaM increased TBF to 153.8 ml/100 g/min after 3 days and to 153.9 ml/100 g/min after 10 days of treatment, compared with 94.0 ml/100 g/min in untreated controls. Cerebral blood flow did not change after EaM treatment. NNM increased TBF but also showed a tendency to increase cerebral blood flow. E2 increased TBF, whereas cerebral blood flow was unchanged. EaM resulted in a rapid reduction in tumor weight from 230 mg in untreated animals to 146 mg after 3 days of treatment. EaM induced an early transient fragmentation of deoxyribonucleic acid in glioma but not in the normal brain. Neither NNM nor E2 affected tumor weight. CONCLUSION: EaM increases TBF in the BT4C rat glioma model with a concomitant rapid antitumoral effect. The increase in TBF could partially be induced by an estrogen-like action of EaM, but the rapid cytotoxic effect of the drug is obviously attributed to the intact EaM compound. This cytotoxic effect might be attributable to the induction of programmed cell death. PMID- 9218313 TI - Impairment of the modulatory role of nitric oxide on the endothelin-1-elicited contraction of cerebral arteries: a pathogenetic factor in cerebral vasospasm after subarachnoid hemorrhage? AB - OBJECTIVE: Nitric oxide (NO) and endothelin-1 (ET-1) are two endothelium-derived factors probably involved in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage (SAH). Our aim was twofold, i.e., to ascertain whether endothelial and nonendothelial NO modulates the contractile response of cerebral arteries to ET-1 and to analyze whether this relationship might be impaired after experimental SAH. METHODS: Rings of middle cerebral artery from goats in the control group and from goats with SAH were set up for isometric tension recordings. SAH was induced 3 days before the experiments by infusion of 10 ml of autologous arterial blood through a catheter previously inserted into the subarachnoid space (basal cistern). In goats in the control group, the response to ET-1 was obtained as follows: 1) in control arteries (unrubbed and nonincubated arteries); 2) in rubbed arteries (arteries in which the endothelium was mechanically removed); 3) during incubation with NG-nitro-L-arginine (L NOArg) alone or plus L- or D-arginine; and 4) in rubbed arteries plus incubation with L-NOArg. In goats with SAH, that response was obtained in control arteries, rubbed arteries, and during incubation with L-NOArg. Specimens of middle cerebral artery were processed for transmission electron microscopy study. RESULTS: In goats in the control group, ET-1 elicited concentration-dependent contraction of the middle cerebral artery that was significantly potentiated after endothelium denudation or during incubation with L-NOArg. The latter effect was reversed by L arginine but not by D-arginine. Combined endothelium denudation and incubation with L-NOArg produced a contractile response to ET-1 significantly higher than that induced by each treatment separately. Hyperreactivity to ET-1 was observed in goats with SAH. Endothelium denudation did not alter the enhanced response to ET-1, but it was further significantly increased after incubation with L-NOArg. CONCLUSION: These results demonstrate that an ET-1-NO interaction exists in control cerebral arteries in such a way that endothelial and nonendothelial NO partially counteract the contractile response to ET-1 and that although SAH did not modify the effect of nonendothelial NO, the absence of endothelial NO after SAH may contribute to the hyperreactivity of cerebral arteries to ET-1 and, thereby, to the development of cerebral vasospasm. PMID- 9218314 TI - Do spinal meningiomas penetrate the pial layer? Correlation between magnetic resonance imaging and microsurgical findings and intracranial tumor interfaces. AB - OBJECTIVE: To study the relationships between spinal dura-arachnoid and tumor cord interfaces in spinal meningiomas and to investigate whether a disruption of the pial layer and penetration of the tumor in the spinal cord occurs. METHODS: Fifteen patients with histologically proven meningiomas underwent magnetic resonance imaging (MRI) preoperatively. All patients underwent microsurgery. The histological characteristics of the tumors were compared with MRI and microsurgical findings. RESULTS: At surgery, the peritumoral hypointense rim revealed by MRI in 10 of 15 patients corresponded to a well-defined cerebrospinal fluid-containing space confined between the outer arachnoidal layer and the inner leptomeningeal layer. In those patients in whom the hypointense peritumoral rim was absent, the inner layer was either difficult to identify or clearly absent, and the blood vessels were extremely adherent to the tumor, requiring a more cautious dissection. Penetration of the tumors through disruption of the pial surface was not documented. CONCLUSION: Previous anatomic and electron microscopy studies demonstrated, in human spinal meninges, the presence of an intermediate layer attached to the inner aspect of the arachnoid, extending laterally over the dorsal surface of the spinal cord and arborizing over the nerve roots and blood vessels. The intermediate layer is not present in human cerebral leptomeninges. The presence/absence of this layer might explain the hypointense rim detected by MRI and might also explain why no penetration and no peritumoral edema is observed in spinal meningiomas as compared with intracranial meningiomas. PMID- 9218315 TI - Yi-Cheng Zhao: a founder of neurosurgery in China. AB - Yi-Cheng Zhao was trained in neurosurgery at the Montreal Neurological Institute by Wilder Penfield in 1938. This article presents Zhao's great contributions to the development of neurosurgery in China. He set up the first independent neurosurgical departments in Tianjin (1952) and in Beijing (1954). A basic research unit for neurosurgery, Beijing Neurosurgical Institute, was also established in 1960 under Zhao's insistent efforts. To raise the clinical level, he emphasized subspecialization of neurosurgery and divided the service into tumor, trauma, pediatrics, and miscellaneous groups. He was also enthusiastic about training neurosurgeons. More than 200 students have been trained by him, and most of them have set up centers in different cities in China. At present, the Beijing Neurosurgical Institute is the largest neurosurgical research unit. It plays an important role in the development of Chinese neurosurgery. Zhao devoted nearly 40 years to neurosurgery and died in 1974. The chinese Neurosurgical Association has honored Zhao as "a founder of neurosurgery in China." PMID- 9218316 TI - Benjamin Winslow Dudley and early American trephination for posttraumatic epilepsy. AB - Benjamin Winslow Dudley (1785-1870) was a Kentucky frontier surgeon who received basic medical education in the United States and extensive surgical training in Europe. He returned to Lexington to become a dominant figure and the most prominent surgical teacher in the Mississippi Valley. Written evidence of Dudley's operative accomplishments are sparse, but he seems to have combined the finest French (Dominique Jean Larrey, Guillaume Dupuytren) and British (Henry Cline, John Abernethy, Astley Cooper) surgical training with conservative and thoughtful patient selection. His operative endeavors in the preantiseptic era included trephination for posttraumatic epilepsy in six patients (1819-1832). This was the largest recorded series of such cases, and it stimulated other American surgeons to trephine for relief of posttraumatic seizures. Trephination for decompression and debridement was undertaken at the site of original injury to remove the cause of "cerebral excitement" and restore "corporeal and intellectual function." Dudley considered this a safe operation in "cautious, firm, and intelligent hands." He thought crowded urban hospitals were unsafe and attributed his better surgical results to the clean, rural Kentucky air. Dudley's achievement is contrasted with other Early American preantiseptic trephinations for posttraumatic epilepsy. PMID- 9218317 TI - Maffucci's syndrome associated with a cranial base chondrosarcoma: case report and literature review. AB - OBJECTIVE AND IMPORTANCE: Our objective was to study the diagnosis and management of this rare condition. A review of the literature concerning chondrosarcomas related to Maffucci's syndrome is reported. Cause and management are discussed. CLINICAL PRESENTATION: We report a case of Maffucci's syndrome associated with a cranial base chondrosarcoma. To our knowledge, only five similar cases have been reported in the literature. The differential diagnosis between Ollier's disease and Maffucci's syndrome and the causes of these conditions are not clear. INTERVENTION: An 18-year-old female patient presented with a giant tumor involving the posterior fossa, clivus, middle fossa, and cavernous sinus. The lesion could be totally removed through a transzygomatic approach. The histological diagnosis was chondrosarcoma. It was confirmed by immunohistochemical studies. There were no postoperative complications. CONCLUSION: Maffucci's syndrome is a rare clinical condition that presents difficulties concerning its diagnosis and management. It is characterized by the presence of multiple enchondromas and cutaneous hemangiomas. Intracranial chondrosarcomas may be associated with this syndrome. Immunohistochemical studies are necessary to differentiate chondrosarcomas from chordomas. The treatment of choice for cranial base chondrosarcomas is total removal of the lesion. Total removal may be very difficult to achieve because of the involvement of neurovascular structures. Alternative therapies, such as proton beam radiosurgery, should be considered. In this case, radical removal of the tumor was possible using a transzygomatic approach. Gross total removal of large cranial base chondrosarcomas is possible, but a longer follow-up period is necessary to ascertain that radical resection was achieved. PMID- 9218318 TI - Eosinophilic granuloma of the clivus: case report, follow-up of two previously reported cases, and review of the literature on cranial base eosinophilic granuloma. AB - OBJECTIVE AND IMPORTANCE: To our knowledge, this is the first reported case of the use of stereotactic radiotherapy for an eosinophilic granuloma (EG) of the clivus. We report follow-up information on two previously reported cases and suggest a management plan for this rare lesion. CLINICAL PRESENTATION: We report the case of a 4.5-year-old boy who presented with a complete abducens palsy on the right with an associated head turn. A computed tomographic scan of his head revealed a lytic lesion on that side, and magnetic resonance imaging showed the mass to be of low intensity on T1-weighted images and of high intensity on T2 weighted images with heterogeneous enhancement. INTERVENTION: A transnasal stereotactic biopsy was performed, revealing an EG. The patient was treated with stereotactic radiotherapy, and he became symptom-free with radiographic resolution of his lesion. Reviewing the literature, we found 13 series with 87 cases of EG in the petrous portion of the temporal bone. EG in the cranial base occurring outside of the temporal bone or in the temporal bone and extending intracranially is, however, quite rare, with only nine other cases reported, two of them clival. CONCLUSION: These findings suggest a classification schema in which cranial base EG lesions be grouped with either the more common extracranial petrous temporal bone lesions or the very rare intracranial lesions. Although there are few cases in the literature, treatment results indicate that clival EG, and perhaps all intracranial cranial base EGs, be treated by a biopsy alone, followed by surgery or stereotactic radiotherapy if there is an incomplete resolution of the symptoms or if there is a recurrence. PMID- 9218319 TI - Primary intracranial metatypical basal cell carcinoma: case report. AB - OBJECTIVE AND IMPORTANCE: A case of primary intracranial metatypic basal cell carcinoma in a 20-year-old man is described. CLINICAL PRESENTATION: A 20-year-old man presented with palsies of the left cranial nerves VI through XII, including complete facial and vestibulocochlear nerve palsy and signs of cerebellar dysfunction, which included left-sided brachydiadochokinesis and nystagmus when looking to the left. There was no evidence of extracranial tumor manifestation. Imaging showed a tumor located in the left pyramidal bone, filling the left cerebellopontine cistern and compressing the brain stem with an extension into the middle cranial fossa as far as the internal carotid artery. INTERVENTION: Subtotal tumor removal was accomplished by a combined neurosurgical otolaryngological procedure through a transpetrosal approach. A histopathological examination revealed a metatypical basal cell carcinoma. Postoperatively, a total dose of 60 Gy of radiation therapy was administered over a period of 6 weeks. CONCLUSION: Although it is rare, primary intracranial basal cell carcinoma should be considered in the differential diagnosis of tumors of the temporal bone. PMID- 9218321 TI - Two unusual cases of multiple dural arteriovenous fistulas. AB - OBJECTIVE AND IMPORTANCE: Two patients with dural arteriovenous fistulas involving the transverse sinus and superior sagittal sinus are described, with a focus on the unique type of venous drainage of the fistula. CLINICAL PRESENTATION: Both patients presented with papilledema and progressive visual disturbance. Angiography and magnetic resonance imaging showed that the fistulas involving the superior sagittal sinus had a dilated venous channel, separate from the sinus lumen, located within the wall of the sinus. INTERVENTION: Transvenous embolization of the venous channel of the fistula, proximal to its drainage into the superior sagittal sinus, resulted in closure of the fistula and restoration of the superior sagittal sinus function. The clinical symptoms were reversed; the symptoms are believed to have reflected venous hypertension in the superior sagittal sinus, resulting from the shunted flow and interfering with normal venous drainage. CONCLUSION: This unique type of dural arteriovenous fistula may be a variant, occurring in the developmental process of the fistula. It is significant clinically because transvenous embolization can be used to close the fistula and restore sinus function. PMID- 9218320 TI - Trochlear nerve schwannomas occurring in patients without neurofibromatosis: case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: Despite their predilection for sensory nerves, intracranial schwannomas have been reported in a number of mixed and purely motor cranial nerves, especially in association with Type 2 neurofibromatosis. We report the rare occurrence of a trochlear nerve schwannoma in a patient without neurofibromatosis and review 17 other case reports from the literature. CLINICAL PRESENTATION: A 35-year-old woman presented with an 8-week history of evolving left hemiparesis, bilateral bulbar paresis, and out-of-character emotional lability. INTERVENTION: She underwent a left temporal craniotomy and a subtemporal, transtentorial approach to the tentorial hiatus, with complete excision of a cisternal trochlear nerve schwannoma. CONCLUSION: Postoperative complications included temporary oculomotor and abducens nerve palsies and temporary right hemiparesis and mild expressive dysphasia, which were resolved at 23-month follow-up. Preoperative symptoms and signs completely resolved, but a postoperative complete trochlear nerve palsy required inferior oblique myectomy for correction of diplopia. A review of the literature showed no preoperative trochlear nerve involvement in at least 45% of cases. The tumor is isointense on T1- and T2-weighted magnetic resonance images and enhances brightly with gadolinium. The most frequently used approach for surgical excision is the subtemporal approach, and the tumor is almost always totally excised. Long-term follow-up suggests recovery of preoperative deficit, and persisting or new trochlear nerve palsy is the rule. PMID- 9218322 TI - Severe rhabdomyolysis with good recovery in a patient with head injury: case report. AB - OBJECTIVE AND IMPORTANCE: We report a case of head injury, in which a hyperosmolar state evolved during the course of treatment, complicated by severe rhabdomyolysis and acute renal failure, which subsequently resulted in a good recovery after intensive supportive treatment. To our knowledge, such high levels of creatine kinase in a patient with head injury and rhabdomyolysis have not been reported. CLINICAL PRESENTATION AND INTERVENTION: A 19-year-old male patient with head injury sustained a compound fracture of the frontal region. He received a hyperosmolar agent to treat brain edema and developed a hyperosmolar state and diabetes insipidus 1 day after the accident. There were no obvious associated injuries at physical examination. After admission to the intensive care unit, the patient developed myoglobinuria and rhabdomyolysis; serum creatine kinase was elevated to a peak of 650,000 IU/L. Four days later, acute renal failure was noted. The patient's myoglobinuria and rhabdomyolysis gradually declined, and he eventually recovered from acute renal failure after supportive treatment and dialysis. CONCLUSION: We postulate that the hyperosmolar state of the patient was the major cause of his severe rhabdomyolysis. Associated hypokalemia and hypophosphatemia are also predisposed to rhabdomyolysis. The most serious complication in rhabdomyolysis is acute renal failure, but most patients who receive supportive treatment and can survive despite the complications can expect to have normal renal function restored. PMID- 9218324 TI - Acrylic cranioplasty with alginate molding: technical note. AB - OBJECTIVE: Acrylic cranioplasty for cranial defects is a common neurosurgical procedure that is performed when the original bone flap becomes infected or is unusable for other reasons. We developed a simple technique to produce a complete copy of the original bone flap from acrylic, using alginate impression material as a mold. METHODS: Alginate impression material was used to form a mold of the patient's original bone flap. Methylmethacrylate was then placed inside the mold to create an exact duplicate of the bone flap. The acrylic flap was sterilized with gamma irradiation and used for cranioplasty. Two patients who had cranial defects secondary to infections of their craniotomy bone flaps underwent cranioplasty with this technique. RESULTS AND CONCLUSION: A perfect copy of the patient's original bone flap was easily and quickly created with this technique, and excellent cosmetic results were obtained. Operative time was shortened because the prosthesis was preformed before the operation. This technique can also be used to mold large or complex cranial defects as long as the original bone flap is available and there is no major cranial remodeling around the defect. PMID- 9218323 TI - Craniocephalic disproportion with increased intracranial pressure and brain herniation: a new clinical syndrome in anemic patients: report of two cases. AB - OBJECTIVE AND IMPORTANCE: We describe a new clinical syndrome in two patients with chronic anemia. The major manifestation of the syndrome is herniation of the brain resulting in death caused by longstanding craniocephalic disproportion. The disproportion was caused by extreme thickening of the cranium because of erythroid hyperplasia. CLINICAL PRESENTATION: Two patients with known chronic anemia presented with chronic increase in intracranial pressure with acute deterioration resulting in brain herniation. INTERVENTION: Despite maximum medical therapy, both patients died as a result of uncontrollable increase in intracranial pressure. CONCLUSION: Patients with chronic anemia presenting with progressive headaches should be monitored for this newly described clinical phenomenon. PMID- 9218325 TI - Partial pediculectomy in the treatment of lumbar spinal stenosis: technical note. AB - OBJECTIVE: We note an additional pathological condition associated with lumbar spinal stenosis that may be responsible for significant postoperative pain. Recognizing that nerve roots are stretched around hypertrophic pedicles in some cases of spinal stenosis, we have altered our surgical management of these cases to address what may be a previously unrecognized but significant anatomic pathological finding. SURGICAL TECHNIQUE: After ipsilateral posterior bony decompression of the spinal canal, the nerve root is examined as it courses around the pedicle. If the root appears stretched, the medial part of the pedicle is removed using first a diamond bit and then a curet. The nerve root is retracted and protected during this procedure. RESULTS: Inspection of the root after partial pediculectomy frequently reveals lateral movement of the root into space previously occupied by the pedicle. Anatomically, the nerve is better decompressed and free of obstruction. This technique adds little time to the overall duration of the operation. CONCLUSION: Anatomic evidence obtained through intraoperative examination and preoperative imaging techniques indicates that partial pediculectomy may play a role in the treatment of some cases of lumbar stenosis. PMID- 9218326 TI - Removal of a lateral disc herniation with malleable endoscopic forceps: technical note. AB - OBJECTIVE: The use of "seeing" endoscopic malleable pituitary forceps in the removal of a lateral herniated lumbar disc is evaluated. METHODS: A malleable pituitary forceps with an attached endoscope was used to explore the neuroforamina and remove laterally herniated disc fragments without a lateral approach or disruption of the facet joint. RESULTS: In the described case, endoscopic forceps provided easy localization and removal of the disc fragments. CONCLUSION: Because this instrument was used successfully in this case, further evaluation of the use of the endoscopic pituitary forceps should be made. PMID- 9218327 TI - Wilder Penfield (1891-1976). PMID- 9218329 TI - New jet irrigation bipolar system. PMID- 9218328 TI - Surgical anatomy of the anterior cervical spine: the disc space, vertebral artery, and associated bony structures. PMID- 9218330 TI - Computer modeling of intracranial saccular and lateral aneurysms for the study of their hemodynamics. PMID- 9218331 TI - Extent of medial temporal resection on outcome from anterior temporal lobectomy: a randomized prospective study. PMID- 9218332 TI - Isolated cerebral hypothermia by single carotid artery perfusion of extracorporeally cooled blood in baboons. PMID- 9218333 TI - Nomenclature of carbohydrates (recommendations 1996). PMID- 9218334 TI - Thioglycosides as glycosyl donors in oligosaccharide synthesis. PMID- 9218336 TI - Sugars and nucleotides and the biosynthesis of thiamine. PMID- 9218337 TI - Molecular architecture of polysaccharide helices in oriented fibers. PMID- 9218339 TI - Sexual activity and contraceptive use among children entering out-of-home care. AB - This study explored the prevalence of reported sexual activity of a cohort of children entering out-of-home care and the ability of selected factors to explain reported sexual activity and use or nonuse of contraceptives. It found that children as young as age 8 reported sexual activity, and that more than one-third of the children age 8 to 18 reported being sexually active. Of those who were sexually active, more than one-third were not using contraceptives. Using logistic regression, five variables are identified as having importance in explaining sexual activity. Two variables had some limited ability to explain contraceptive use. Implications of these findings are discussed and suggestions for policy and practice are made. PMID- 9218340 TI - Family empowerment: one outcome of cooperative preschool education. AB - Although longitudinal evaluations of academic preschools have consistently demonstrated significant and substantial long-term benefits for children, the effect of the preschool experience on parents who are required to participate has not been examined. This article reviews a qualitative investigation that explored family empowerment as one outcome of cooperative preschool education. The article highlights the personal growth and acquisition of skills that occurred during the midphase of the empowerment process. PMID- 9218341 TI - Outcomes for infants exposed in utero to illicit drugs. AB - The study reported here sought to determine whether substance-exposed infants who are maltreated have a higher risk of out-of-home placement than substance-exposed children who are not abused and rejected, as well as a higher risk of death than children in the general population. In a sample of 513 infants born at a Chicago medical center from 1985 through 1990, 480 (93.6%) had complete sociodemographic data available for analysis. Identifying data were used to search the Illinois death registry and a computerized central registry of child abuse reports. Both out-of-home placement and death were distinctly more likely if children had been exposed to drugs and maltreated. Such children should be closely followed. PMID- 9218342 TI - The experience of family foster care in Malawi: a preliminary investigation. AB - This preliminary investigation into family foster care in Malawi set out to provide information on the Malawi government family foster care structure primarily through the qualitative experience of foster caregivers. A semistructured interview schedule was used to interview 24 foster parents, who were selected from a government family foster care register, in two regions of Malawi. The study revealed that cultural factors influenced various aspects of family foster care, ranging from the caregivers' decision to foster children to the caregivers' determination not to disclose to the children that they were fostered. Social and economic factors also played a role, particularly with regard to some shortcomings in service delivery. PMID- 9218343 TI - Shelters for runaway and homeless youths: capacity and occupancy. AB - Data from a nationally representative sample of shelters for runaway and homeless youths (N = 160) were analyzed to determine shelter capacity, occupancy, and occupancy ratios. Analysis focused in particular on occupancy ratios by funding status, shelter size, metropolitan statistical area (MSA), season, and day of the week. PMID- 9218345 TI - Liquid chromatographic determination of cocaine, benzoylecgonine, and cocaethylene in whole blood and serum samples with diode-array detection. AB - An extraction with Bond Elut Certify solid-phase extraction (SPE) columns is developed for the isolation of cocaine, benzoylecgonine, and cocaethylene from whole blood and serum followed by reversed-phase liquid chromatography with diode array detection. Two internal standards (2'-methylbenzoylecgonine and 2' methylcocaine) with close structural resemblance to benzoylecgonine (a carboxylic acid) and to the two esters, cocaine and cocaethylene, are used in the analytical procedure. A thorough evaluation of this SPE and a comparison with different liquid-liquid extractions clearly show the superiority of the SPE. A linear response (correlation coefficient greater than 0.998) over a broad concentration range (0.025-5.0 micrograms/mL) is obtained. The sensitivity, specificity, precision (coefficients of variation less than 4.9% for within-day reproducibility and less than 5.3% for total reproducibility), and accuracy of the method are excellent for each analyte. Forensic blood samples from people suspected of cocaine abuse are analyzed and show the usefulness of the method, even for degraded postmortem samples. PMID- 9218344 TI - Detection of tricyclic antidepressants in whole blood by headspace solid-phase microextraction and capillary gas chromatography. AB - A simple method for the extraction of four tricyclic antidepressants from whole blood by headspace solid-phase microextraction (SPME) is presented. The whole blood samples contain four drugs (amitriptyline, chlorimipramine, imipramine, and trimipramine) and are heated at 100 degrees C in a septum-capped vial in the presence of distilled water and NaOH solution; a polydimethylsiloxane-coated SPME fiber is exposed to the headspace of the vial to allow adsorption of the drugs before capillary gas chromatography (GC) with flame-ionization detection. The headspace SPME-GC produces intense peaks for each drug with very little background noise. Recoveries of the four drugs by the present method are 5.3 12.9%. The calibration curves for the drugs show linearity in the range of 31 1000 ng/0.5 mL. The detection limits of each drug are 16-25 ng/0.5 mL. Imipramine is detectable from rat blood 5 h after oral administration of imipramine (500 mg/kg body weight); the concentration is 1.44 +/- 0.209 micrograms/mL. PMID- 9218346 TI - Two-dimensional proton chemical-shift imaging of human muscle metabolites. AB - Large lipid signals and strong susceptibility gradients introduced by muscle-bone interfaces represent major technical challenges for in vivo proton MRS of human muscle. Here, the demonstration of two-dimensional proton chemical-shift imaging of human muscle metabolites is presented. This technique utilizes a chemical shift-selective method for water and lipid suppression and automatic shimming for optimal homogeneity of the magnetic field. The 2D1H CSI technique described facilitates the acquisition of high-spatial-resolution spectra, and allows one to acquire data from multiple muscle groups in a single experiment. A preliminary investigation utilizing this technique in healthy adult males (n = 4) revealed a highly significant difference in the ratio of the creatine to trimethylamine resonance between the fast and slow twitch muscle groups examined. The technique is robust, can be implemented on a commercial scanner with relative ease, and should prove to be a useful tool for both clinical and basic investigators. PMID- 9218347 TI - Optimization of the water-PRESS pulse sequence and its integration into pulse sequences for studying biological macromolecules. AB - In this paper, the recently developed "Water-PRESS" method of water suppression [W. .S. Price and Y. Arata (1996), J. Magn. Reson. B 112, 190] in which homospoil pulses are used to manipulate the effects of radiation damping on the water resonance and thereby selectively alter the effective relaxation times of the water resonance with respect to the solute (e.g., biological macromolecules) resonances is further developed and applied. In the present work, methods for optimization in terms of degree of water suppression and in temporal terms (important for the application of Water-PRESS to multidimensional experiments) are considered so that recycle delays of less than 2.3 s (including the acquisition time) are possible. Also, a simple modification which allows the observation of solute resonances with relaxation times similar to that of the water resonance is presented. Finally, the inclusion into more complicated pulse sequences is also discussed. Experimental examples using aqueous samples of lysozyme and immunoglobulin are given. Compared to most other NMR water suppression techniques, this method is extremely simple to implement and optimize and does not require accurately calibrated RF pulses or perfect lineshape. PMID- 9218348 TI - Effect of long TE on T1 measurement in STEAM progressive saturation experiment. PMID- 9218349 TI - Kinematics and functional morphology of aquatic feeding in Australian snake necked turtles (Pleurodira;Chelodina). AB - Head kinematics during aquatic feeding of the Australian long-necked turtle (Chelodina) were studied by means of high speed video recordings. Buccal expansion was assessed by calculation of elliptical cross-sectional surfaces. Further, displacements of head, carapace, and prey in the earth bound frame, of the prey relative to the center of the gape, and of the head relative to the carapace were determined. Rates of change (velocities) of all these variables were calculated. These data are combined with information on the osteology and myology of the head. The robust development of the large hyobranchial apparatus, the massive intercornuatus muscle, and the presence of the branchiosquamosus muscle were related to aquatic feeding skills. Head kinematics are variable in amplitude and relative timing, but proceed always in a rostrocaudal sequence. According to their effect on the prey, two components are distinguished in the process of expansion. The first compensates for head/body movements (compensatory suction). The second causes distinct acceleration of water and prey (inertial suction). The latter component is mainly driven by the abduction of the second branchial arch. In spite of largely different structural solutions, optimal feeding conditions as deduced for suction in feeding fishes are also employed by Chelodina. This further promotes the assumption that hydrodynamics constrain evolutive solutions for aquatic feeding. PMID- 9218351 TI - Killer toxins of certain yeast strains have potential growth inhibitory activity on gram-positive pathogenic bacteria. AB - The killer yeast strains which are encountered most frequently among species in the genera Saccharomyces, Candida, Hansenula, Pichia and Kluyveromyces (ten killer strains in toto) were tested for the activity of their toxins on the growth of some pathogenic bacteria (four Gram-positive and six Gram-negative strains) in a test in which purified toxins were not required. Neither toxins of the killer Saccharomyces cerevisiae and Pichia membranefaciens strains were active against the bacterial strains. In contrast the killer toxins of Hansenula anamola, Hansenula mrakii, Candida tropicalis, Kluyveromyces drosphilarum and Kluyveromyces lactis showed potential growth inhibitory effects on Gram-positive pathogenic bacteria. Thus, yeast killer toxins were found to be active only on Gram-positive bacterial cell types. PMID- 9218352 TI - A Staphylococcus aureus enterotoxin-B induced type 1 immediate hypersensitivity reaction in a goat. AB - A Nigerian dwarf goat intended for the production of antibodies to staphylococcal enterotoxin-B exhibited classical signs of a type 1 anaphylactic reaction 2 to 3 min after parenteral introduction of this antigen. The exquisite sensitivity demonstrated in concert with the pathognomonic signs appeared to make the breed and species an excellent model for the further study and interpretation of this syndrome in domestic food animals. PMID- 9218353 TI - Alternative splicing of the rabbit beta-globin pre-mRNA in vivo after transient transfection of myoblast and myotube cells. AB - The relationship between preferences among alternative 5' splice sites and their sequences was investigated using as model mouse myoblasts and myotubes after transient transfection with the rabbit beta-globin gene. The preferences for the use of two different 5' splice sites, acting with different efficiencies to direct splicing in vivo are reported. The predominant selection of the upstream splice site has been shown in normal mouse myoblasts. In the case of differentiated myotubes the downstream splice was 1.4 times better used. The results indicate that there were differences in the preferences for the use of the two alternative splice sites between non-differentiated and terminally differentiated cells, within the same-cell line. PMID- 9218354 TI - Antibacterial and antifungal activity of aromatic constituents of essential oils. AB - Five aromatic constituents of essential oils (cineole, citral, geraniol, linalool and menthol) were tested for antimicrobial activity against eighteen bacteria (including Gram-positive cocci and rods, and Gram-negative rods) and twelve fungi (three yeast-like and nine filamentous). In terms of antibacterial activity linalool was the most effective and inhibited seventeen bacteria, followed by cineole, geraniol (each of which inhibited sixteen bacteria), menthol and citral aromatic compounds, which inhibited fifteen and fourteen bacteria, respectively. Against fungi the citral and geraniol oils were the most effective (inhibiting all twelve fungi), followed by linalool (inhibiting ten fungi), cineole and menthol (each of which inhibited seven fungi) compounds. PMID- 9218355 TI - Production, purification and characterization of an extracellular alpha-amylase enzyme isolated from Aspergillus flavus. AB - Filamentous fungi isolated from cereals were screened for their ability to produce alpha-amylase (1,4-alpha-glucan glucanohydrolase, EC 3.2.1.1). A selected strain identified as Aspergillus flavus showed high enzymatic activity. A single extracellular alpha-amylase was purified to homogeneity by a starch adsorption method. The molecular weight (M(r)) of the A. flavus alpha-amylase was approximately 75,000 +/- 3,000 by polyacrylamide gel electrophoresis (PAGE) and that of the subunit was approximately 75,000 +/- 3000 SDS-PAGE. The optimal activity of the purified enzyme was achieved at pH 7.0 and 30 degrees C. K+ ions increased the alpha-amylase activity, but Mg2+ did not greatly affect enzyme activity. Mn2+, Zn2+, Cu2+ and Fe3+ ions strongly inhibited the enzyme activity. The products of hydrolysis of native starch by the A. flavus enzyme were mainly glucose as well as unidentified oligosaccharides. PMID- 9218356 TI - Quantitative bioanalysis utilizing high-performance liquid chromatography/electrospray mass spectrometry via selected-ion monitoring of the sodium ion adduct [M+Na]+. AB - A high-performance liquid chromatography (HPLC)/electrospray mass spectrometric method for quantitative determination of a compound in dog plasma was developed and validated via the selected-ion monitoring of the electrospray-generated [M+Na]+ adduct of the compound. The plasma samples were acidified with HCl and then extracted with methyl tert-butyl ether. The reconstituted extracts were injected into an HPLC/positive-ion electrospray ionization mass spectrometry system. The HPLC mobile phase consisted of acetonitrile, water, formic acid (3 mM) and sodium acetate (0.3 mM). This composition of mobile phase provided the optimum electrospray condition for the formation of the [M+Na](+)-ion. This work demonstrates that the addition of sodium acetate into the HPLC mobile phase and the subsequent selected-ion monitoring of the sodium ion adduct of the analyte is a viable approach in quantitative bioanalysis. The facile formation of the sodium ion adduct of the analyte, which does not contain functional groups that are known to be strong proton acceptors, appears to be a function of the particular electrospray instrument used. PMID- 9218357 TI - Low picogram determination of Ro 48-6791 and its major metabolite, Ro 48-6792, in plasma with column-switching microbore high-performance liquid chromatography coupled to ion spray tandem mass spectrometry. AB - A coupled liquid chromatography/tandem mass spectrometry assay was developed for simultaneous determination of Ro 48-6791 and its secondary amine metabolite in human plasma samples with a quantification limit for both compounds of 1 pg/mL using a 1 mL plasma aliquot. The method exploits the enhanced mass sensitivity of a microbore (300 microns i.d.) reversed-phase capillary column coupled to an ion spray probe combined with tandem mass spectrometry. A straightforward column switching system was utilized to focus the analytes onto a microbore trapping column following solid-phase extraction of a 50 microL plasma sample extract from liquid/liquid extraction. Backflushing of the retained analytes from the trapping column onto the microbore capillary column provided the requisite high peak concentration for high sensitivity. The inter-assay precision and accuracy for Ro 48-6791 and its metabolite, at 10 pg/mL, were found to be 3.4%, and 105%, and 9.1%, and 99.9%, respectively. The calibration curves were linear over the range 1 to 1000 pg/mL. The method proved to be sufficiently rugged for analysis of samples. PMID- 9218358 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry as a rapid screening method to detect mutations causing Tay-Sachs disease. AB - Matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) has been used as a rapid method for the detection of human genetic polymorphisms. In particular, the mutations in the human HEXA gene that cause the infantile Tay Sachs disease have been studied using MALDI-MS to demonstrate the feasibility of this technique for use in clinical and diagnostic analysis. The protocols involved in this approach include, polymerase chain reaction for the amplification of the mutation site from buccal cell DNA, followed by restriction enzyme digestion of the amplified regions of the template cells. The products of amplification and digestion were studied using MALDI-MS. MALDI-MS experiments are shown to provide essentially the same information as obtained from gel electrophoresis but orders of magnitude faster. PMID- 9218359 TI - Persistence of MS-2 and PRD-1 bacteriophages in an ultrapure water system. AB - The persistence of bacteriophages MS-2 and PRD-1 was evaluated in tap water, in reverse osmosis (RO) permeate, and in three locations within an ultrapure water system; ultrapure samples included pre- and post-UV sterilization and post-mixed bed ion exchange tank. The inactivation rates for MS-2 were calculated as log10 reduction per hour and per day: k = -(log 10 Ct/C0)/t. PRD-1 was found to persist with no significant loss of infectivity in all water purity environments evaluated. Inactivation of MS-2 was dependent on water quality and pH. Short-term inactivation rates for chlorinated tap water, post-RO, pre-UV, post-UV and post ion exchange sample locations were 0.028, 0.455, 0.231, 0.191 and 0.168 log10 h 1, respectively. Long-term inactivation rates for chlorinated tap water, post-RO, pre-UV, post-UV and post-ion exchange sample locations were 0.485, 0.911, 0.605, 0.632 and 0.684 log10 day-1, respectively. Since phages were found to remain intact as well as to lyse in the ultrapure water environment, the phages have the potential to contaminate the ultrapure water environments of the microelectronics, pharmaceutical and power generation industries in both colloidal and dissolved form. Further work is proceeding to generate standardized and cost-effective methods to detect viruses in water environments. PMID- 9218360 TI - Microbial diversity: the importance of exploration and conservation. AB - Microbial diversity is fundamental to maintenance and conservation of global genetic resources. As extreme environments are explored, the richness of microbial diversity is increasingly evident. Measures must be taken to estimate, record, and conserve microbial diversity, not only to sustain human health but also to enrich the human condition globally through wise use and conservation of genetic resources of the microbial world. PMID- 9218361 TI - Effects of medium composition and nutrient limitation on loss of the recombinant plasmid pLG669-z and beta-galactosidase expression by Saccharomyces cerevisiae. AB - The effects of medium composition, nutrient limitation and dilution rate on the loss of the recombinant plasmid pLG669-z and plasmid-borne beta-galactosidase expression were studied in batch and chemostat cultures of Saccharomyces cerevisiae strain CGpLG. The difference in growth rates between plasmid-free and plasmid-containing cells (delta mu) and the rate of segregation (R) were determined and some common factors resulting from the effect of medium composition on plasmid loss were identified. Glucose-limited chemostat cultures of CGpLG grown on defined medium were more stable at higher dilution rates and exhibited delta mu-dominated plasmid loss kinetics. Similar cultures grown on complex medium were more stable at lower dilution rates and exhibited R-dominated plasmid loss kinetics. Overall plasmid stability was greatest in phosphate limited chemostat cultures grown on defined medium and was least stable in magnesium-limited cultures grown on defined medium. delta mu decreased and R increased with increased dilution rate, irrespective of medium composition. Increased plasmid loss rates at high or low dilution rates would appear to be characteristic of loss kinetics dominated by R or delta mu, respectively. Growth of glucose-limited chemostat cultures on complex medium decreased delta mu values but increased R values, in comparison to those cultures grown on defined medium. Any increased stability that a complex medium-induced reduction of delta mu may have conferred was counteracted by an increased R value. Increased beta galactosidase productivity was correlated with increased plasmid stability only in glucose-limited chemostat cultures grown on defined medium and not in those grown on complex medium. Previous studies have yielded contrasting responses with regard to the effect of dilution rate on recombinant plasmid loss from S. cerevisiae. Our findings can account for these differences and may be generally valid for the stability of similar yeast plasmid constructs. This information would facilitate the design of bioprocesses, where recombinant plasmid instability results in reduced culture productivity. PMID- 9218362 TI - Production methods for rabies vaccine. PMID- 9218363 TI - A home electrocardiography and blood pressure telemonitoring system. AB - A home electrocardiography (ECG) and blood pressure telemonitoring system for cardiac patients was installed in the Macau region. The monitoring centre was established in the emergency unit at the Government Hospital of Macau. The first users were 10 cardiovascular patients selected by a physician. The average age of these users was 61 years (range 30-78). The results of a three-month trial showed that the system was easy to operate and technically reliable. It was found to be helpful for cardiac patients. The most significant problem during the trial was electrical noise from the ECG electrodes. PMID- 9218364 TI - Telepsychiatry: 'tele' yes, but what about the 'psychiatry'? AB - To investigate what is lost or gained in a psychiatric evaluation when it takes place via telepsychiatry we compared the interrater reliability between two psychiatrists interviewing 63 subjects in an observer/interviewer split configuration in telepsychiatry and same-room settings. The measures used were the BPRS and interviewer ratings from a semi-structured interview. Patients also rated their experience. There were some clear differences between the telepsychiatry and same-room evaluations. Despite these variations, diagnoses were as reliably made by telepsychiatry. Patient acceptance of telepsychiatry was high. PMID- 9218365 TI - Interpersonal communications and telemedicine: hypotheses and methods. AB - Demonstration interviews between a psychiatrist who has used videolinks for a range of clinical interactions and a simulated patient facilitate a better understanding of interpersonal communication, both face to face and mediated. This might allow users to be trained to maximize the use of the available communications technology and provide a source of more rational reassurance for the technophobic professional. PMID- 9218366 TI - Telemedicine for airline passengers, seafarers and islanders. AB - Airline passengers, seafarers and islanders are three different examples of people who can benefit from telemedicine. However, the peculiar characteristics of each group require different applications. In 60 years of activity, the CIRM has assisted more than 37,000 patients. In the 10 years from 1986 to 1996 the Centre provided radio medical assistance to 7647 patients, of whom 6981 (91.3%) were sailors, 642 (8.4%) were people living in isolated areas (small italian islands) with few medical facilities, while only 24 patients (0.3%) were airline passengers. In the same period, the telecommunication service received or transmitted almost 80,000 medical messages. PMID- 9218367 TI - A pilot study in medical education using interactive television. AB - Medical students in the United Arab Emirates do not receive postmortem teaching. This is because postmortems are not normally carried out, for cultural reasons. In order to address this problem a collaborative project was established between the medical schools of Aberdeen University and the United Arab Emirates University to evaluate the feasibility, acceptability and effectiveness of telepathology teaching. A videoconferencing link was established between the UK and the Middle East using ISDN at a transmission speed of 384 kbit/s. Although some technical problems relating to line continuity were encountered, the results relating to feasibility, acceptability and effectiveness were very positive. Although expensive, this form of teaching may still be cost-effective in relation to the benefits. PMID- 9218368 TI - A referrer and patient evaluation of a telepsychiatry consultation-liaison service in South Australia. AB - A study was carried out to describe the patient population assessed by a telepsychiatry consultation-liaison service in rural South Australia, and to assess the referrers' and patients' satisfaction rating with this service. The study was performed in two parts, with retrospective and prospective components. The author completed a semi-structured interview for each patient (n = 75) with a Brief Psychiatric Rating Scale (BPRS) for the prospective group (n = 32). A questionnaire was also sent to all referrers seeking an evaluation of the usefulness of the telepsychiatry interview in terms of assessment and management recommendations and outcome. Patients from the prospective group were sent a questionnaire examining their evaluation of the usefulness of the interview in terms of assessment and management recommendations, and difficulties with the technology. The patient population was characterized by high rates of affective disorder and personality disorder, and high indices of developmental disturbance. Referrers reported high rates of satisfaction with the service. Nursing staff rated the service more positively than general practitioners. The usefulness for assessment was rated more highly than for management. PMID- 9218369 TI - Is there a role for telemedicine in an urban environment? AB - Semi-structured interviews were carried out to determine the expectations of potential participants in a feasibility study of teleconsulting. General practitioners produced high positive scores in all areas. Hospital consultants produced high scores but were more critical of technical performance and administrative arrangements. Patient satisfaction with teleconsulting was generally good. The results suggested that teleconsulting was both feasible and acceptable to the users. PMID- 9218370 TI - Expert pathology consultation through the Internet: melanoma versus benign melanocytic tumours. AB - Twenty consecutive cases of melanocytic lesions were chosen from the archives of the archives of the institute of Anatomic Pathology at Santa Chiara Hospital, Trento. Representative images were acquired at a spatial resolution of 512 x 512 pixels, saved in IPEG format and delivered to the remote pathologist by multimedia internet electronic mail. Six cases were diagnosed as benign melanocytic lesions by the local pathologist. Of the 20 cases transmitted, each with an average of 5.3 images, the remote pathologist suggested a diagnosis of malignancy in nine cases while 10 cases were thought to be benign. In one case the images were not considered sufficient for diagnosis. Overall, the diagnostic agreement between local and remote pathologist was 79% (kappa = 0.58, P = 0.002). This preliminary study suggest that telepathology by internet electronic mail can be a valuable tool for remote consultation in dematopathology, as well as for other diagnostic fields where expert consultation is necessary. PMID- 9218371 TI - A cost analysis of a tele-oncology practice. AB - Costs were monitored for three different types of oncology practice: a telemedicine clinic and a fly-in outreach clinic, both held in rural areas, and a traditional clinic held in a city hospital. Total expenses were calculated over the year May 1995 to April 1996. The average cost per telemedicine visit was $812. The average cost per outreach clinic visit was $897. Flying in oncology support for this practice was therefore about 10% more costly than telemedicine. While the outreach cost may have been inappropriately high due to a slow start-up phase, it was still less expensive during this period to be seen via telemedicine. For comparison, the average cost per traditional oncology clinic visit was $149. However, this figure does not take into account the costs of access to a city-based service by rural patients. PMID- 9218372 TI - Continuous automated telecare assessment of the elderly. AB - An automated scheme is proposed to assess the ability of elderly people to live alone in the community. It employs an Enhanced Activities of Daily Living index based on a computerized questionnaire form. In addition, a number of low-cost sensors have been developed which provide electronic measures of certain activities; these can provide additional inputs to the assessment form using telemetry. The sensors are capable of measuring a wide range of functional performance, thus providing the means of continuously and objectively assessing a patient's condition following hospitalization. PMID- 9218373 TI - High-quality television links for home-based support for the elderly. AB - Since 1991, 17 elderly people have been connected via a broadband video communication system to a telecare centre. The new services used videophones based on domestic television sets, including set-top boxes with cameras and a microphone. The services had the overall aim of promoting the ability of elderly and mobility-impaired people to live independently and to reduce the demand on social service resources required to implement this. Service components included: remote response to emergencies; active information and care; information and assistance service; remote care on demand; remote access to expertise (counselling); training and exercise service; and support for carers. Some 14,000 calls have been made. The important qualitative aspects were: clients' satisfaction; replacement of direct social contacts; privacy and data protection; and improving the effectiveness of social services. For a successful market implementation of video-based social support and telecare services, it is essential to integrate them into existing outpatient services. PMID- 9218374 TI - Realtime ultrasound diagnosis over a wide-area network (WAN) using off-the-shelf components. AB - A trial system was developed for relaying realtime ultrasound images from an obstetric referral centre, the Maitland Hospital, to a tertiary-care centre, the John Hunter Hospital in Newcastle. The sites were approximately 30 km apart and connected by a microwave link at 2 Mbit/s. The pilot study demonstrated the feasibility of realtime ultrasound transmission using commonly available components. PMID- 9218375 TI - Telecare equipment in the home. Issues of intrusiveness and control. AB - Telecare in the home offers substantial benefits to users. However, the manner of its development and technological configuration will determine the extent to which it is acceptable and will meet clinical and social objectives. There are important issues of intrusiveness and control. An ethical framework for telecare is needed. PMID- 9218376 TI - Evaluating the alliance in videolink teletherapy. AB - The use of videoconferencing in psychotherapy remains largely unexplored. Videoconferencing compromises the range and quality of interactional information and thus might be expected to affect the working alliance (WA) between client and therapist, and consequently the process and outcome of therapy. A single case study exploring the effect of videoconferencing on the development of the WA in the psychological treatment of a female-male transsexual is described. The self rated Working Alliance Inventory (WAI) was used to measure client and therapist perceptions of the WA after each session over 10 sessions of eclectic therapy conducted over a videolink. The serial WAI measurements charting the development of the WA in 4 cases of 10-session, face-to-face therapy by Horvath and Marx were used as a quasi-control. Therapist and client impressions of teletherapy are described. WAI scores were essentially similar to the face-to-face control group except for lower client-rated bond subscale scores. It is suggested that client personality factors accounted for this difference and that videoconferencing did not impair the development of an adequate working alliance or successful therapeutic outcome. PMID- 9218377 TI - Report of a national neurosurgical teleradiology system. AB - An emergency neurosurgical teleradiology system was initially installed in two referring hospitals in ireland to transmit images to the neurosurgical department in Cork. It was subsequently expanded to six major referring hospitals transmitting to both neurosurgical departments in ireland serving the entire population of 3.5 million people, effectively becoming a national teleradiology system. The system was based on PCs interconnected by leased data circuits and ISDN. The network was operational 24 hours a day. Over 750 emergency computerized tomography scans were transmitted and transmission failures occurred in only 6% of cases. We conclude that current PC technology can be used to form a peer-to peer wide-area network upon which a robust emergency teleradiology system can be based. PMID- 9218378 TI - Lessons learned during the INSURRECT project (INteractive SURgical Teaching at REmote CeNtres). AB - The INSURRECT project began in 1993 as a collaboration between six UK universities in distance teaching of undergraduate surgery. The first year was spent in testing the network and preparing the course material. This was followed by a two-year pilot teaching course. During this phase, 108 teaching sessions were conducted, involving more than 1300 students in all. It was found that successful teaching depended on increasing the amount of audience interaction was much as possible and transmitting high-quality video pictures. Although the time taken to deliver material by interactive video was greater than for conventional lectures, both students and teachers responded favourably to the project. PMID- 9218379 TI - Education and training of practice nurses. AB - Seventeen nurses in eight rural general practices participated in a distance education project. Low-cost videoconferencing equipment was assessed for its suitability in two training sessions, concerning asthma and travel immunization. The intended learning outcomes were reached and although initially apprehensive, the nurses quickly became accustomed to the medium. Videoconferencing has now become an accepted part of in-service training. Technical reliability remains the most important problem. PMID- 9218380 TI - Telemedicine demonstration projects in the Western Pacific. AB - Telemedicine demonstration projects in the Western Pacific are attempting to provide distance medical consulting and distance medical education for the isolated health-care workforce of the region. The Picasso Phone system is a low cost telemedicine system which requires only the standard telephone network for its operation, needs little additional equipment and is user-friendly. The demonstration projects have shown the utility of the system in both international, inter-island and on-island communications. All demonstration activities used the various existing island telephone systems. With the adaptation of such a user-friendly telemedicine system as the Picasso Phone system to the Peacesat public-interest educational network, the capacity of the telemedicine network will expand to include isolated islands with Peacesat capacity which do not have access to the commercial island telephone systems. PMID- 9218381 TI - Telemedicine and the New Children's Hospital (Royal Alexandra Hospital for Children). AB - Telemedicine is an important factor in the future strategy of the Royal Alexandra Hospital for Children. As a state, national and international centre of excellence, it is the hospital's role to assist in the development of the best child health services. An important aim is to provide easy access to a full range of paediatric services through the provision of a comprehensive telemedicine service, which encompasses videoconferencing and tele-imaging, and positions the child as the centre of service provision. Experience so far suggests that in Australia, as in other countries, the adoption of telemedicine may outstrip the ability of the legislative and administrative frameworks to keep pace. Thus, the enablers appear to be cultural and technological while the obstacles are rooted in the way in which health systems are financed and administered. PMID- 9218382 TI - Comprehensive standardized ophthalmic telemedicine. AB - The Electrodiagnostic Neurophysiological Automated Analysis (ENAA) telematic system was developed in the EU EUREKA project. To validate the system 2500 electrodiagnostic tests were administered in a standardized manner during a three year period. The tests were performed on 70 normal subjects and 500 patients in five laboratories in three European countries. The data were transmitted to the Bristol Telematic Electrodiagnostic Centre in the Bristol Eye Hospital. Data from normal subjects were not significantly different between laboratories. Data from patients were reported upon and the conclusions transmitted to the place of origin. The system provided the remote consultant with multimedia data, including medical images such as colour fundus photography and angiography, video and sound. PMID- 9218383 TI - Telepsychiatry, the satellite system and family consultation. AB - A pilot telepsychiatry session was conducted with the US Department of Defense Satellite Communication System. The subjects were a family incompletely divorced many years before. There were two satellite interviews with this family. Bringing together all members of the original family so that questions could be addressed as to what happened when the children were very young unblocked a 13-year-old communication problem. PMID- 9218384 TI - Remote consultation for computerized tomography and magnetic resonance studies by means of teleradiology--experience at the Singapore General Hospital. AB - A teleradiology link was established between Singapore General Hospital in Singapore and Stanford University in California, USA. Over eight months, a total of 28 cases (involving 27 magnetic resonance investigations and three computerized tomography scans) were transmitted by ISDN to California. Our initial experience with teleradiology for remote consultation was encouraging, although the data transmission cost was higher than we anticipated. however, costs could be reduced by using data compression. Long-distance telecommunication charges continue to fall, so intercontinental teleradiology of this type may be financially viable in future. PMID- 9218385 TI - Effect size and experimental power analysis in a paediatric cardiology telemedicine system. AB - A telemedicine system was installed between the University of North Carolina Hospitals and the New Hanover Regional Medical Center. It allowed the transmission of neonatal echocardiograms for immediate reporting. During a six month study period the system was used for the interpretation of 110 echocardiograms from 48 babies. There were 38 babies studied in a retrospective control period. Hospital length of stay decreased by an average of six days in the telemedicine group, representing an annual saving of some $1.3 million. However, these apparent differences were not significant (P = 0.2) and a power analysis suggested that a sample size of some 600 would have been necessary. PMID- 9218386 TI - Telemedicine in emergency home care--the 'Shahal' experience. AB - Shahal serves over 40,000 cardiac, pulmonary and blood pressure subscribers. The system combines emergency home care and telemedicine in a patient-initiated system geared towards the prevention of cardiac and pulmonary complications. About 150,000 calls are received per year. The median time from onset of symptoms to a call for help is 44 min. It is a unique system which has been shown to facilitate improved home health-care control, enabling patients to manage their own health condition and providing them with a higher quality of life and enhanced peace of mind. PMID- 9218387 TI - Towards a comprehensive telecardiology monitoring centre for community-based services. AB - In a pilot study of primary-care telecardiology, 2563 consultations were carried out over 18 months. Following teleconsultation, 2076 patients (81%) were found to be suitable for management entirely by the general practitioner, without the need for referral to hospital. The system identified 487 patients (19%) with cardiac problems who required either admission to hospital or outpatient assessment. There was a resultant saving of referrals to hospital accident and emergency departments. Extension of the telecardiology service to include tele echocardiography may result in faster access to diagnosis and better management of patients in heart failure, improving patients' quality of life and reducing hospitalization. PMID- 9218388 TI - The contribution of telemedicine to cardiology. AB - The Telemedicine Centre at Sismanoglion Hospital of Athens is connected to seven primary-care units (HCCs) on the mainland and six HCCs on Aegean islands. Telemedicine activity from 1992 to 1995 was reviewed. During the study period, the data relating to 1947 cardiac patients were transmitted through the telemedicine network: 681 (35%) of the patients presented with cardiological problems and 333 (17%) of them had an urgent cardiac event. The telemedicine network brought a number of benefits, including better access to cardiology specialists, improved decisions about patient transportation, a reduction in isolation and continuing professional education. PMID- 9218389 TI - Telemedicine applications for home health care. AB - Home health care in the USA is one of the most rapidly growing segments of the health-care market. Telehealth seeks to reduce some of the inefficiencies of home health care in various ways, including replacing certain nursing visits with video visits, collecting vital-signs data remotely, improving medication compliance and patient education. The use of telehealth in home health-care settings will provide a means of interacting in a client-centred manner, promoting client autonomy through education and improved communications. PMID- 9218390 TI - Home telenursing in Kansas: patients' perceptions of uses and benefits. AB - Elderly individuals involved in a home telenursing project were studied. The project nurses provided home health services from 'telenursing cockpits' located in three separate sites in Kansas. A cable television-based interactive video system was used to transmit video pictures at 30 frames/s, with 288 horizontal lines of resolution. During phase 1 of the study, interview data were collected from 22 subjects (4 men, 18 women). During phase 2, the original participants were contacted but only 9 (1 man, 8 women) were still receiving home health services. Contrary to expectations, the technology was not an important issue for the participants. They did not express any particular worry or excitement about it. Nor did they describe difficulties in adapting to its use. Use of telemedicine technology did not appear to have any negative effects on communication. The results suggests that further thought needs to be given to defining clearly the purpose and goals of telemedicine projects. PMID- 9218391 TI - Use of neural networks in telemedical monitoring of divers. AB - An artificial neural network (ANN) has been developed to predict and classify the risk of medical disorders for certain dive profiles. The telemedical data applied to the ANN represent different physiological and physical parameters that are transmitted acoustically from a diver to a remote receiver. The telemetered data include dive time, maximum depth and decompression stop times and depths. Preliminary tests demonstrated the successful performance of the ANN where classical mathematical and statistical methods had failed due to the complex nature and variability of the parameters involved. PMID- 9218392 TI - Preliminary results from the Northern Ireland arms of the UK Multicentre Teledermatology Trial: effect of camera performance on diagnostic accuracy. AB - The diagnostic accuracy of realtime teledermatology was measured using two different video cameras. One camera was a relatively low-cost, single-chip device (camera 1), while the other was a more expensive three-chip camera (camera 2). The diagnosis obtained via the videolink was compared with the diagnosis made in person. Sixty-five new patients referred to a dermatology clinic were examined using camera 1 followed by a standard face-to-face consultation. A further 65 patients were examined using camera 2 and the same procedure applied. Seventy-six per cent of conditions were correctly diagnosed by telemedicine using camera 2 compared with 59% using camera 1. A working differential diagnosis was obtained in 12% of cases using camera 2 compared with 17% using camera 1. The percentage of 'no diagnosis', wrong and missed diagnoses was halved using camera 2 compared with camera 1. These results suggest that the performance of camera 2 was superior to that of camera 1 for realtime teledermatology. PMID- 9218393 TI - Telemedicine delivery to developing countries. AB - This paper highlights current activities with regard to telemedicine activities in and for developing countries. The paper reviews: the preparation of a telemedicine report by a study group of the International Telecommunication Union (ITU), the aim of which is to provide recommendations and guidelines for developing countries; the formation of the European Telemedicine Collaboration Group (ETCG), which is undertaking telemedicine pilot projects in developing countries; and telemedicine delivery via inmarsat, which is coordinating production of the ITU report and is a participant in the ETCG. PMID- 9218394 TI - Tele-education: the virtual medical laboratory. AB - The virtual medical laboratory (VML) was conceived to provide an Internet accessible resource, offering access for clinicians and scientists to an invaluable data archive at the institute of Laryngology and Otology, London. The Institute is home to the largest collection of temporal bone, laryngeal, skull and sinus sections in Europe. The skull and sinus collections include an extensive section consisting of animal material. These were contributions from zoos around the world. Over the last 50 years, samples have been carefully sectioned and stained by specialized technicians to produce histology slices of most regions of the head and neck. The aim of the project is to create a virtual medical laboratory, which will provide access to archived histological material as well as computerized tomography and magnetic resonance data. Central to this aim is the reconstruction of the internal anatomy of the temporal bone from two dimensional histology slices, to create three-dimensional views that can be used for anatomical simulation and surgical training in otolaryngology. State-of-the art three-dimensional reconstruction and rendering technology allows us to develop such a model. Computer-generated simulation could be made available to all hospitals in which otolaryngology is practised, via digital communication networks. We aim to develop core technology in our own specialty that is applicable to other fields of higher education, which have not been exposed to such modern teaching modalities. This has the potential to become an invaluable teaching resource for anatomists, surgeons and other scientists. PMID- 9218395 TI - A portable digital imaging system in dermatology: diagnostic and educational applications. AB - Digital photographs were taken by a trainee dermatologist of the presenting lesions of 100 unselected, consecutive new patients. For the 38 patients presenting with rashes there was clinical disagreement in only four cases (10%). For the 62 patients with tumours there was clinical disagreement in three cases (3.8%). In a further three cases both clinicians agreed on a differential diagnosis which was subsequently disproved by histological findings and clinical progress. The study demonstrated that an affordable, low-resolution, fixed-focus digital camera with close-up lenses could provide diagnostically useful images suitable for telediagnosis in dermatology. PMID- 9218396 TI - Remote, mobile telemedicine: the satellite transmission of medical data from Mount Logan. AB - The purpose of this investigation was to demonstrate the potential of remote, mobile telemedicine during a four-week, high-altitude mountaineering expedition to Mount Logan, Canada's highest summit. Using a mobile satellite terminal and a laptop computer (both powered by a photovoltaic solar panel), ECG tracings and blood pressure measurements, in addition to colour images, short-segment video and audio clips were transmitted during the course of the ascent. The data were transmitted via a mobile communications satellite to a ground station in Ottawa, a distance of over 4000 km. The data were then transferred to the public switched data network and delivered to the University of Ottawa Heart Institute for analysis. Similarly, data were transmitted from the ground station to the expedition team on Mount Logan throughout the ascent. Using this technique, medical diagnosis and emergency care can be facilitated in extreme and isolated locations lacking a telecommunications infrastructure. Such technology has applications in developing countries, disaster response efforts, remote civilian and military operations, and in space operations. PMID- 9218397 TI - A telemedical record model. AB - Medical record issues appear to be the weak link in telemedicine. A telemedical record model is proposed which would fit both a managed-care and fee-for-service health-care environment. In this model, the originator of the teleconsultation referral is the party responsible for creation of the record, since that is the initial point of patient contact. The telemedical record model prevents duplication of information gathering and provides a consistent way to document teleconsultations. Telemedical record policies and procedures should be in place before starting a telemedicine programme, to ensure consistency in the documentation of doctor-patient encounters. PMID- 9218398 TI - Too much food or too little exercise? PMID- 9218400 TI - Unravelling the mystery of obesity. PMID- 9218399 TI - An oil rig worker presenting with acute gout. PMID- 9218401 TI - Why is losing weight so hard? PMID- 9218402 TI - Managing insulin-dependent patients. PMID- 9218403 TI - Impotence in the diabetic patient. PMID- 9218404 TI - A protocol for thyrotoxicosis. PMID- 9218405 TI - Spotting diabetic retinopathies. PMID- 9218406 TI - New thinking on the treatment of rhinitis. PMID- 9218407 TI - Where next for you--and the MRCGP? PMID- 9218408 TI - A slimming group outing to the chip shop. PMID- 9218409 TI - Sequences and topology: Predicting evolution. PMID- 9218410 TI - Web alert: Nucleic acids Sequences and topology. PMID- 9218411 TI - Platelet-activating factor acetylhydrolases. PMID- 9218412 TI - SH3-mediated Hck tyrosine kinase activation and fibroblast transformation by the Nef protein of HIV-1. AB - Tyrosine kinases of the Src family are regulated via their Src homology 2 (SH2) and SH3 domains. The Nef protein of human immunodeficiency virus-1 (HIV-1) has previously been shown to bind with high affinity and specificity in vitro to the SH3 domain of Hck, a Src family member expressed primarily in myeloid cells. However, the effect of Nef on Hck activity in living cells is unknown. Here we show that Rat-2 fibroblasts co-expressing Hck and Nef rapidly developed transformed foci, whereas control cells expressing either protein alone did not. Nef formed a stable complex with Hck and stimulated its tyrosine kinase activity in vivo. Mutagenesis of the Nef proline-rich motif essential for SH3 binding completely blocked complex formation, kinase activation, and transformation, indicating that the Nef SH3-binding function is required for its effects on Hck. These results provide direct evidence that SH3 engagement is sufficient to activate a Src family kinase in vivo and suggest that Hck may be activated by Nef in HIV-infected macrophages. PMID- 9218413 TI - A new metabolic link. The acyl carrier protein of lipid synthesis donates lipoic acid to the pyruvate dehydrogenase complex in Escherichia coli and mitochondria. AB - Lipoic acid is an essential enzyme cofactor that requires covalent attachment to its cognate proteins to confer biological activity. The major lipoylated proteins are highly conserved enzymes of central metabolism, the pyruvate and alpha ketoglutarate dehydrogenase complexes. The classical lipoate ligase uses ATP to activate the lipoate carboxyl group followed by attachment of the cofactor to a specific subunit of each dehydrogenase complex, and it was assumed that all lipoate attachment proceeded by this mechanism. However, our previous work indicated that Escherichia coli could form lipoylated proteins in the absence of detectable ATP-dependent ligase activity raising the possibility of a class of enzyme that attaches lipoate to the dehydrogenase complexes by a different mechanism. We now report that E. coli and mitochondria contain lipoate transferases that use lipoyl-acyl carrier protein as the lipoate donor. This finding demonstrates a direct link between fatty acid synthesis and lipoate attachment and also provides the first direct demonstration of a role for the enigmatic acyl carrier proteins of mitochondria. PMID- 9218414 TI - A combinatorial approach defines specificities of members of the caspase family and granzyme B. Functional relationships established for key mediators of apoptosis. AB - There is compelling evidence that members of the caspase (interleukin-1beta converting enzyme/CED-3) family of cysteine proteases and the cytotoxic lymphocyte-derived serine protease granzyme B play essential roles in mammalian apoptosis. Here we use a novel method employing a positional scanning substrate combinatorial library to rigorously define their individual specificities. The results divide these proteases into three distinct groups and suggest that several have redundant functions. The specificity of caspases 2, 3, and 7 and Caenorhabditis elegans CED-3 (DEXD) suggests that all of these enzymes function to incapacitate essential homeostatic pathways during the effector phase of apoptosis. In contrast, the optimal sequence for caspases 6, 8, and 9 and granzyme B ((I/L/V)EXD) resembles activation sites in effector caspase proenzymes, consistent with a role for these enzymes as upstream components in a proteolytic cascade that amplifies the death signal. PMID- 9218415 TI - Insight into lipid surface recognition and reversible conformational adaptations of an exchangeable apolipoprotein by multidimensional heteronuclear NMR techniques. AB - Apolipophorin III (apoLp-III) from the insect Manduca sexta is a 166-residue (Mr 18,340) member of the exchangeable apolipoprotein class that functions to stabilize lipid-enriched plasma lipoproteins. In the present study, we present the secondary structure and global fold of recombinant apoLp-III derived from three-dimensional heteronuclear NMR spectroscopy experiments. Five discrete alpha helical segments (21-30 residues in length) with well defined boundaries were characterized by four NMR parameters: medium range nuclear Overhauser enhancement contacts between proton pairs, chemical shift index, coupling constants, and amide proton exchange rates. An antiparallel arrangement of helical segments has been obtained based on the long range interhelical nuclear Overhauser enhancement contacts. The NMR solution structure reveals a globular, up and down helix bundle organization similar to that of Locusta migratoria apoLp-III (Breiter, D. R., Kanost, M. R., Benning, M. M., Wesenberg, G., Law, J. H., Wells, M. A., Rayment, I., and Holden, H. M. (1991) Biochemistry 30, 603-608). However, a short helix (comprised of 5 amino acids) has been identified in the region between helix 3 and helix 4. This helix is postulated to play a role in lipid surface recognition and/or initiation of binding. Our results also indicate the existence of buried polar and charged residues in the helix bundle, providing a structural basis for the relatively low stability of apoLp-III in its lipid-free state. It is suggested that the intrinsic low stability of lipid-free apoLp-III may be important in terms of its ability to undergo a reversible, lipid binding-induced, conformational change. This study underscores the striking resemblance in molecular architecture between insect apoLp-III and the N-terminal domain of human apolipoprotein E. The potential for application of NMR techniques to studies of the exchangeable apolipoproteins, possibly in their biologically active, lipid-associated state, has broad implications in terms of our understanding of the molecular basis of their physiological functions. PMID- 9218416 TI - Autoinduction of retinoic acid metabolism to polar derivatives with decreased biological activity in retinoic acid-sensitive, but not in retinoic acid resistant human breast cancer cells. AB - Previous studies have shown that all-trans-retinoic acid (RA) inhibits in vitro proliferation of hormone-dependent human breast cancer cells but not the growth of hormone-independent cells. Here we report on RA metabolism in breast cancer cells as examined by high performance liquid chromatography analysis and found a correlation with sensitivity to growth inhibition by RA. RA-sensitive T-47D and MCF-7 cells exhibited high rate metabolism to polar metabolites, whereas RA resistant MDA-MB-231 and MDA-MB-468 cells metabolized RA to a much lesser extent, and almost no polar metabolites could be detected. The high metabolic rate in RA sensitive cells appears to be the result of autoinduction of RA metabolism, whereas RA-resistant cells showed no such induction of metabolism. We observed furthermore that transfection with retinoic acid receptor-alpha expression vectors in RA-resistant MDA-MB-231 cells resulted in increased RA metabolism and inhibition of cell proliferation. Metabolism of RA, however, seems not to be required to confer growth inhibition of human breast cancer cells. The biological activity of the polar metabolites formed in RA-sensitive cells was found to be equal or lower than that of RA, indicating that RA itself is the most active retinoid in these cells. Together our data suggest that RA-sensitive cells contain mechanisms to activate strongly the catabolism of RA probably to protect them from the continuous exposure to this active retinoid. PMID- 9218417 TI - Apoptosis induced by gliotoxin is preceded by phosphorylation of histone H3 and enhanced sensitivity of chromatin to nuclease digestion. AB - The fungal toxin gliotoxin induces apoptotic cell death in a variety of cells. Apoptosis induced in thymocytes by gliotoxin is rapid, and DNA fragmentation is observable within 4 h treatment. Apoptosis induced by gliotoxin is calcium independent and unaffected by protein synthesis inhibitors. We have previously shown that gliotoxin results in phosphorylation of a 16.3-kDa protein within 10 min treatment of thymocytes. Here we show that this protein is histone H3 and phosphorylation occurs on Ser-10. Cyclic AMP levels and activity of protein kinase A (PKA) are raised in cells treated with gliotoxin. Apoptosis is inhibited by genistein which also inhibits PKA and histone H3 phosphorylation. Apoptosis is also inhibited by a number of specific inhibitors of PKA suggesting apoptosis induced by gliotoxin is modulated by this kinase. The agents forskolin and cholera toxin do not induce rapid phosphorylation of H3 although some increase in phosphorylation of H3 does occur after 8 h with these agents. Forskolin and cholera toxin also induce apoptosis but over a longer time course than gliotoxin. In all cases levels of apoptosis correlate with degree of H3 phosphorylation. Cells treated with gliotoxin show an early sensitivity to micrococcal nuclease and DNase I digestion indicating a functional relationship between DNA fragmentation and H3 phosphorylation. PMID- 9218418 TI - Role of the GLUT 2 glucose transporter in the response of the L-type pyruvate kinase gene to glucose in liver-derived cells. AB - Twenty-six different hepatoma cell lines established from cancer-prone transgenic mice exhibited a close correlation between expression of the GLUT 2 glucose transporter and activation of the L-type pyruvate kinase (L-PK) gene by glucose, as judged by Northern blot analyses and transient transfection assays. The L-PK gene and a transfected L-PK construct were silent in GLUT 2(+) cells and active in GLUT 2(-) cells cultured in glucose-free medium. Transfection of GLUT 2(-) cells with a GLUT 2 expression vector restored the inducibility of the L-PK promoter by glucose, mainly by suppressing the glucose-independent activity of this promoter. Culture of GLUT 2(-) cells, in which the L-PK gene is constitutively expressed, in a culture medium using fructose as fuel selected GLUT 2(+) clones in which the L-PK gene responded to glucose. The expression of the L-PK gene in GLUT 2(-) cells cultured in the absence of glucose was correlated with a high intracellular glucose 6-phosphate (Glu-6-P) concentration while under similar culture conditions Glu-6-P concentration was very low in GLUT 2(+) cells. Consequently, a role of GLUT 2 in the glucose responsiveness of glucose-sensitive genes in cultured hepatoma cells could be to allow for Glu-6-P depletion under gluconeogenic culture conditions. In the absence of GLUT 2, glucose endogeneously produced might be unable to be exported from the cells and would be phosphorylated again to Glu-6-P by constitutively expressed hexokinase isoforms, continuously generating the glycolytic intermediates active on the L-PK gene transcription. PMID- 9218419 TI - Stimulation of intracellular Ca2+ levels in human neutrophils by soluble immune complexes. Functional activation of FcgammaRIIIb during priming. AB - Soluble immune complexes bind to unprimed neutrophils and generate intracellular Ca2+ transients but fail to activate the NADPH oxidase. Following priming of the neutrophils with either tumor necrosis factor alpha or granulocyte-macrophage colony-stimulating factor, stimulation of the cells with the soluble immune complexes leads to an enhanced Ca2+ signal and significant secretion of reactive oxidants. The enhanced Ca2+ signal observed in primed neutrophils results from the influx of Ca2+ from the external environment and is partly sensitive to tyrosine kinase inhibitors. This is in contrast to the Ca2+ signal observed in unprimed neutrophils, which arises from the mobilization of intracellular stores. When the surface expression of FcgammaRIIIb on primed neutrophils was decreased either through incubation with Pronase or phosphoinositide-specific phospholipase C, the extra enhanced Ca2+ mobilization seen in primed cells was significantly lowered, while the initial rise in intracellular Ca2+ was unaffected. Depletion of FcgammaRIIIb had no significant effect on the Ca2+ transients in unprimed neutrophils. Cross-linking FcgammaRII, but not FcgammaRIIIb, induced increases in intracellular Ca2+ in unprimed neutrophils, while cross-linking either of these receptors increased Ca2+ levels in primed neutrophils. The FcgammaRII-dependent intracellular Ca2+ rise in primed cells was unaffected by incubation in Ca2+-free medium, whereas the FcgammaRIIIb-dependent transient was significantly decreased when Ca2+ influx was prevented in Ca2+-free medium supplemented with EGTA. Cross linking either FcgammaRII or FcgammaRIIIb in primed or unprimed cells failed to stimulate substantial levels of inositol 1,4,5-trisphosphate production. These results indicate that following stimulation of primed neutrophils with soluble immune complexes the enhanced Ca2+ mobilization observed is the result of a functional activation of the glycosylphosphatidylinositol-linked FcgammaRIIIb. PMID- 9218420 TI - Identification of the histidine protein kinase KinB in Pseudomonas aeruginosa and its phosphorylation of the alginate regulator algB. AB - The exopolysaccharide alginate is an important virulence factor in chronic lung infections caused by the bacterium Pseudomonas aeruginosa. Two positive activators for alginate synthesis, algB and algR, are members of a superfamily of response regulators of the two-component regulatory system. AlgB belongs to the NtrC subfamily of response regulators and is required for high-level production of alginate. In this study, an open reading frame encoding a polypeptide of 66 kDa, designated kinB, was identified immediately downstream of algB. The sequence of KinB is homologous to the histidine protein kinase members of two-component regulatory systems. Western blot analysis of a P. aeruginosa strain carrying a kinB-lacZ protein fusion and studies of kinB-phoA fusions indicate that KinB localizes to the inner membrane and has a NH2-terminal periplasmic domain. A KinB derivative containing the COOH terminus of KinB was generated and purified. In the presence of [gamma-32P]ATP, the purified COOH-terminal KinB protein was observed to undergo progressive autophosphorylation in vitro. Moreover, the phosphoryl label of KinB could be rapidly transferred to purified AlgB. Substitutions of the residues conserved among histidine protein kinases abolished KinB autophosphorylation. These results provide evidence that kinB encodes the AlgB cognate histidine protein kinase. PMID- 9218421 TI - Binding of the volatile anesthetic chloroform to albumin demonstrated using tryptophan fluorescence quenching. AB - The site(s) of action of the volatile general anesthetics remain(s) controversial, but evidence in favor of specific protein targets is accumulating. The techniques to measure directly volatile anesthetic binding to proteins are still under development. Further experience with the intrinsic protein fluorescence quenching approach to monitor anesthetic-protein complexation is reported using chloroform. Chloroform quenches the steady-state tryptophan fluorescence of bovine serum albumin (BSA) in a concentration-dependent, saturable manner with a Kd = 2.7 +/- 0.2 mM. Tryptophan fluorescence lifetime analysis reveals that the majority of the quenching is due to a static mechanism, indicative of anesthetic binding. The ability of chloroform to quench BSA tryptophan fluorescence was decreased markedly in the presence of 50% 2,2,2 trifluoroethanol, which causes loss of tertiary structural contacts in BSA, indicating that protein conformation is crucial for anesthetic binding. Circular dichroism spectroscopy revealed no measurable effect of chloroform on the secondary structure of BSA. The results suggest that chloroform binds to subdomains IB and IIA in BSA, each of which contains a single tryptophan. Earlier work has shown that these sites are also occupied by halothane. The present study therefore provides experimental support for the theory that structurally distinct general anesthetics may occupy the same domains on protein targets. PMID- 9218422 TI - Investigation of the pyrimidine preference by the c-Myb DNA-binding domain at the initial base of the consensus sequence. AB - The principal determinant of the pyrimidine preference by the c-Myb DNA-binding domain at the initial base of the consensus sequence was investigated by mutation of both the protein and the DNA base pairs, with analysis by a filter binding assay. Amino acid residue 187 was revealed to interact with the pyrimidine base position, as estimated from our previous complex structure. Unexpectedly, since the pyrimidine preference is retained even in the Gly187 mutant, the principal origin of the base specificity should not occur via the direct-readout mechanism, but by an indirect-readout mechanism, namely in the intrinsic "bendability" of the pyrimidine-purine step of the DNA duplex. A significant but rather small positive base pair roll is detectable in the conformation of DNA in complex with the c-Myb DNA-binding domain. Following the conventional chemical rules of the direct-readout mechanism, amino acid mutagenesis at position 187 yielded several new base preferences for the protein. PMID- 9218423 TI - Targeted replacement of the mouse apolipoprotein E gene with the common human APOE3 allele enhances diet-induced hypercholesterolemia and atherosclerosis. AB - Apolipoprotein (apo) E, a constituent of several lipoproteins, is a ligand for the low density lipoprotein receptor, and this interaction is important for maintaining cholesterol and triglyceride homeostasis. We have used a gene replacement strategy to generate mice that express the human apoE3 isoform in place of the mouse protein. The levels of apoE mRNA in various tissues are virtually the same in the human apoE3 homozygous (3/3) mice and their littermates having the wild type mouse allele (+/+). Total cholesterol and triglyceride levels in fasted plasma from the 3/3 mice were not different from those in the +/+ mice, when maintained on a normal (low fat) chow diet. We found, however, notable differences in the distribution of plasma lipoproteins and apolipoprotein E between the two groups: beta-migrating lipoproteins and plasma apoB100 levels are decreased in the 3/3 mice, and the apoE distribution is shifted from high density lipoproteins to larger lipoprotein particles. In addition, the fractional catabolic rate of exogenously administered remnant particles without apoE was 6 fold slower in the 3/3 mice compared with the +/+ mice. When the 3/3 and +/+ animals were fed a high fat/high cholesterol diet, the 3/3 animals responded with a dramatic increase (5-fold) in total cholesterol compared with the +/+ mice (1.5 fold), and after 12 weeks on this same diet the 3/3 animals developed significantly (at least 13-fold) larger atherosclerotic plaques in the aortic sinus area than the +/+ animals. Thus the structural differences between human APOE3 and mouse ApoE proteins are sufficient to cause an increased susceptibility to dietary-induced hypercholesterolemia and atherosclerosis in the 3/3 mice. PMID- 9218424 TI - Plasma membrane Ca2+ pump isoforms 2a and 2b are unusually responsive to calmodulin and Ca2+. AB - The full-length a and b variants of the rat plasma membrane calcium pump, isoform 2 (rPMCA2a and rPMCA2b), were constructed and expressed in COS-7 cells. To characterize these isoforms, calcium transport was determined in a microsomal fraction. Both rPMCA2a and rPMCA2b had a much higher affinity for calmodulin than the corresponding forms of hPMCA4, and rPMCA2b had the highest affinity among the isoforms that have been tested so far. When analyzed at a relatively high calmodulin concentration, rPMCA2b and, to a lesser extent, rPMCA2a showed higher apparent calcium affinity; i.e. they were more active at lower Ca2+ concentrations than hPMCA4b. This indicates that these two variants of rat isoform 2 will tend to maintain a lower free cytosolic Ca2+ level in cells where they are expressed. Both variants also showed a higher level of basal activity (in the complete absence of calmodulin) than hPMCA4b, a property which would reinforce their ability to maintain a low free cytosolic Ca2+ concentration. Experiments designed to determine the source of the higher apparent Ca2+ affinity of rPMCA2b showed that it came from the properties of the carboxyl terminus, rather than from any difference in the catalytic core. PMID- 9218425 TI - Insulinotropic glucagon-like peptide-1-mediated activation of non-selective cation currents in insulinoma cells is mimicked by maitotoxin. AB - Maitotoxin (MTX) activates a Ca2+-dependent non-selective cation current (ICa-NS) in insulinoma cells whose time course is identical to non-selective cation currents activated by incretin hormones such as glucagon-like peptide-1 (GLP-1), which stimulate glucose-dependent insulin secretion by activating cAMP signaling pathways. We investigated the mechanism of activation of ICa-NS in insulinoma cells using specific pharmacological reagents, and these studies further support an identity between MTX- and GLP-1-activated currents. ICa-NS is inhibited by extracellular application of genistein, econazole, and SKF 96365. This inhibition by genistein suggests that tyrosine phophorylation may play a role in the activation of ICa-NS. ICa-NS is not inhibited by incubation of cells in glucose free solution, by extracellular tetrodotoxin, nimodipine, or tetraethylammonium, or by intracellular dialysis with 4-aminopyridine, ATP, ryanodine, or heparin. ICa-NS is also not significantly inhibited by staurosporine, which does, however, partially inhibit the MTX-induced rise of intracellular Ca2+ concentration. These effects of staurosporine suggest that protein kinase C may not be involved in the activation of ICa-NS but that it may regulate intracellular Ca2+ release. Alternatively, ICa-NS may have a small component that is carried through separate divalent cation-selective channels that are inhibited by staurosporine. ICa-NS is neither activated nor inhibited by dialysis with KF, KF + AlF3 or GTPgammaS (guanosine 5'-O-(3-thiotriphosphate)), suggesting that GTP-binding proteins do not play a major role in the activation of this current. PMID- 9218426 TI - Distinctive functions of Syk and Lyn in mediating osmotic stress- and ultraviolet C irradiation-induced apoptosis in chicken B cells. AB - By taking advantage of the established chicken B cell line, DT40 cells, which do not express tyrosine kinase Syk or Lyn, functional roles of Syk and Lyn in apoptotic response elicited by cellular stress were investigated. DT40 cells underwent apoptosis after hyperosmotic stress. In Syk-deficient DT40 cells, this apoptotic process was significantly enhanced. Ectopic expression of wild type, but not kinase-inactive, porcine Syk in Syk-deficient cells rescued cells from osmotic stress-induced apoptosis, demonstrating that the presence of functionally active Syk is necessary to protect cells from osmotic stress-induced apoptosis. In comparison, there was no effect on osmotic stress-induced apoptosis in Lyn deficient DT40 cells. Interestingly, while Syk was not involved in ultraviolet C (UVC)-induced apoptosis, a deficiency of Lyn rendered cells resistant to UVC irradiation. These observations defined Syk and Lyn as important mediators of apoptosis in DT40 cells in response to osmotic stress and UVC irradiation, respectively. Furthermore, osmotic stress, but not UVC irradiation, could activate c-Jun N-terminal kinase (JNK) in DT40 cells. A deficiency in either Syk or Lyn did not affect the osmotic stress-induced activation of JNK. We, therefore, concluded that Syk and Lyn regulate the apoptotic responses to osmotic stress and UVC irradiation independently of the JNK pathway in DT40 cells. PMID- 9218427 TI - The interaction of the transforming growth factor-betas with heparin/heparan sulfate is isoform-specific. AB - We have undertaken a comparative study of the interaction of the three mammalian transforming growth factor-betas (TGF-beta) with heparin and heparan sulfate. TGF beta1 and -beta2, but not -beta3, bind to heparin and the highly sulfated liver heparan sulfate. These polysaccharides potentiate the biological activity of TGF beta1 (but not the other isoforms), whereas a low sulfated mucosal heparan sulfate fails to do so. Potentiation is due to antagonism of the binding and inactivation of TGF-beta1 by alpha2-macroglobulin, rather than by modulation of growth factor-receptor interactions. TGF-beta2.alpha2-macroglobulin complexes are more refractory to heparin/heparan sulfate, and those involving TGF-beta3 cannot be affected. Comparison of the amino acid sequences of the TGF-beta isoforms strongly implicates the basic amino acid residue at position 26 of each monomer as being a vital binding determinant. A model is proposed in which polysaccharide binding occurs at two distinct sites on the TGF-beta dimer. Interaction with heparin and liver heparan sulfate may be most effective because of the ability of the dimer to co-operatively engage two specific sulfated binding sequences, separated by a distance of approximately seven disaccharides, within the same chain. PMID- 9218428 TI - The acidic region of the factor VIII light chain and the C2 domain together form the high affinity binding site for von willebrand factor. AB - A binding site for von Willebrand factor (vWf) was previously localized to the carboxyl terminus of the C2 domain of the light chain (LCh) of factor VIII (fVIII). The acidic region of the LCh, residues 1649-1689, also controls fVIII.vWf binding by an unknown mechanism. Although anti-acidic region monoclonal antibodies prevent formation of the fVIII.vWf complex, the direct involvement of the acidic region in this binding has not been demonstrated. By limited proteolysis of LCh with Staphylococcus aureus V8 protease, we prepared 14- and 63 kDa LCh fragments, which begin with fVIII residues 1672 and 1795, respectively. Using surface plasmon resonance to measure binding interactions, we demonstrated that the 14-kDa fragment binds to vWf, but its affinity for vWf (Kd 72 nM) was 19 fold lower than that of LCh. This was not due to an altered conformation of the acidic region within the 14-kDa fragment, since its affinity for an anti-acidic region monoclonal antibody was similar to that of LCh. All LCh derivatives lacking the acidic region (thrombin-cleaved LCh, recombinant C2, and 63-kDa fragment) had also greatly reduced affinities for vWf (Kd 564-660 nM) compared with LCh (Kd 3.8 nM). In addition, the similar affinities of these derivatives for vWf indicated that apart from its acidic region, the LCh contains no vWf binding site other than the one within C2. The reduced affinities of the LCh derivatives lacking the acidic region for monoclonal antibody NMC-VIII/5 (epitope, C2 residues 2170-2327) indicated that removal of the acidic region leads to a conformational change within C2. This change is likely to affect the conformation of the vWf binding site in C2, which overlaps the epitope of NMC VIII/5; therefore, the acidic region also appears to be required to maintain the optimal conformation of this vWf binding site. Our results demonstrate that the acidic region and the C2 domain are both directly involved in forming a high affinity binding site for vWf. PMID- 9218429 TI - Characterization of a Goalpha mutant that binds xanthine nucleotides. AB - Several GTP binding proteins, including EF-Tu, Ypt1, rab-5, and FtsY, and adenylosuccinate synthetase have been reported to bind xanthine nucleotides when the conserved aspartate residue in the NKXD motif was changed to asparagine. However, the corresponding single Goalpha mutant protein (D273N) did not bind either xanthine nucleotides or guanine nucleotides. Interestingly, the introduction of a second mutation to generate the Goalpha subunit D273N/Q205L switched nucleotide binding specificity to xanthine nucleotide. The double mutant protein GoalphaD273N/Q205L (GoalphaX) bound xanthine triphosphate, but not guanine triphosphate. Recombinant GoalphaX (GoalphaD273N/Q205L) formed heterotrimers with betagamma complexes only in the presence of xanthine diphosphate (XDP), and the binding to betagamma was inhibited by xanthine triphosphate (XTP). Furthermore, as a result of binding to XTP, the GoalphaX protein underwent a conformational change similar to that of the activated wild type Goalpha. In transfected COS-7 cells, we demonstrate that the interaction between GoalphaX and betagamma occurred only when cell membranes were permeabilized to allow the uptake of xanthine diphosphate. This is the first example of a switch in nucleotide binding specificity from guanine to xanthine nucleotides in a heterotrimeric G protein alpha subunit. PMID- 9218430 TI - Fumonisin B1-induced sphingolipid depletion inhibits vitamin uptake via the glycosylphosphatidylinositol-anchored folate receptor. AB - The folate receptor, like many glycosylphosphatidylinositol-anchored proteins, is found associated with membrane domains that are insoluble in Triton X-100 at low temperature and that are enriched in cholesterol and sphingolipids. Depletion of cellular cholesterol has been shown to inhibit vitamin uptake by this receptor (Chang, W. -J., Rothberg, K. G., Kamen, B. A., and Anderson, R. G. W. (1993) J. Cell Biol. 118, 63-69), suggesting that these domains regulate this process. In this study, the importance of sphingolipids for folate receptor function was investigated in Caco-2 cells using fumonisin B1, a mycotoxin that inhibits the biosynthesis of these lipids. The folate receptor-mediated transport of 5 methyltetrahydrofolate was almost completely blocked in cells in which sphingolipids had been reduced by approximately 40%. This inhibition was dependent on the concentration and duration of the treatment with the mycotoxin and was mediated by the sphingolipid decrease. Neither receptor-mediated nor facilitative transport was inhibited by fumonisin B1 treatment, indicating that the effect of sphingolipid depletion was specific for folate receptor-mediated vitamin uptake. A concurrent loss in the total amount of folate binding capacity in the cells was seen as sphingolipids were depleted, suggesting a causal relationship between folate receptor number and vitamin uptake. These findings suggest that dietary exposure to fumonisin B1 could adversely affect folate uptake and potentially compromise cellular processes dependent on this vitamin. Furthermore, because folate deficiency causes neural tube defects, some birth defects unexplained by other known risk factors may be caused by exposure to fumonisin B1. PMID- 9218431 TI - Characterization of a novel Na+-dependent, guanosine-specific, nitrobenzylthioinosine-sensitive transporter in acute promyelocytic leukemia cells. AB - NB4 cells are the only bona fide in vitro model of human acute promyelocytic leukemia. We have examined cytidine and guanosine transport in this cell line and characterized a novel guanosine-specific transporter. Cytidine transport occurred predominately by equilibrative nitrobenzylthioinosine (NBMPR)-sensitive (es) transport. In the presence of Na+, guanosine at various concentrations accumulated at least 6-fold above equilibrium. The initial rate of guanosine transport in Na+ buffer decreased by 75% with the addition of 1 microM NBMPR and the IC50 for NBMPR inhibition was 0.7 +/- 0.1 nM. Replacement of Na+ with choline also resulted in a 75% decrease in total guanosine transport. The potent inhibition of guanosine transport by NBMPR and the loss of transport in choline suggested that a Na+-dependent NBMPR-sensitive transporter was responsible for the majority of guanosine uptake. This concentrative, sensitive transporter is Na+ dependent with a stoichiometric coupling ratio of 1:1. This novel transporter, referred to as csg, is guanosine-specific with total guanosine transport inhibited by only 50% in the presence of 1 mM competing nucleosides. HL 60, acute myelocytic leukemia cells, do not exhibit csg activity while L1210, murine acute lymphocytic leukemia cells, exhibit csg transport. The presence of the csg transporter suggests an important role for guanosine in particular forms of leukemia and may provide a new target for cytotoxic therapy. PMID- 9218432 TI - Hsp70 prevents activation of stress kinases. A novel pathway of cellular thermotolerance. AB - Harmful conditions including heat shock, oxidative stress, UV, and so forth cause programmed cell death, whose triggering requires activation of the Jun N-terminal kinase, JNK. High levels of Hsp72, a heat-inducible member of Hsp70 family, protect cells against a variety of stresses by a mechanism that is unclear at present. Here we report that elevated levels of Hsp72 inhibit a signal transduction pathway leading to programmed cell death by preventing stress induced activation of JNK. Stress-induced activation of another stress-kinase, p38 (HOG1), is also blocked when the level of Hsp72 is increased. Similarly, addition of a purified recombinant Hsp72 to a crude cell lysate reduced p38 kinase activation, while depletion of the whole family of Hsp70 proteins with a monoclonal antibody enhanced such activation. In addition, we have found that accumulation of abnormal proteins in cells upon incubation with amino acid analogs causes activation of JNK and p38 kinases, which can be prevented by overproduction of Hsp72. Taken together, these data suggest that, in regulation of JNK and p38 kinases, Hsp70 serves as a "sensor" of the build-up of abnormal proteins after heat shock and other stresses. The inhibitory effect of an increased level of Hsp70 on JNK appears to be a major contributor to acquired thermotolerance in mammalian cells. PMID- 9218433 TI - NMR spectroscopic studies of the DNA-binding domain of the monomer-binding nuclear orphan receptor, human estrogen related receptor-2. The carboxyl-terminal extension to the zinc-finger region is unstructured in the free form of the protein. AB - Unlike steroid and retinoid receptors, which associate with DNA as dimers, human estrogen related receptor-2 (hERR2) belongs to a growing subclass of nuclear hormone receptors that bind DNA with high affinity as monomers. A carboxyl terminal extension (CTE) to the zinc-finger domain has been implicated to be responsible for determining the stoichiometry of binding by a nuclear receptor to its response element. To better understand the mechanism by which DNA specificity is achieved, the solution structure of the DNA-binding domain of hERR2 (residues 96-194) consisting of the two putative zinc fingers and the requisite 26-amino acid CTE was analyzed by multidimensional heteronuclear magnetic resonance spectroscopy. The highly conserved zinc-finger region (residues 103-168) has a fold similar to those reported for steroid and retinoid receptors, with two helices that originate from the carboxyl-terminal ends of the two zinc fingers and that pack together orthogonally, forming a hydrophobic core. The CTE element of hERR2 is unstructured and highly flexible, exhibiting nearly random coil chemical shifts, extreme sensitivity of the backbone amide protons to solvent presaturation, and reduced heteronuclear (1H-15N) nuclear Overhauser effect values. This is in contrast to the dimer-binding retinoid X and thyroid hormone receptors, where, in each case, a helix has been observed within the CTE. The implications of this property of the hERR2 CTE are discussed. PMID- 9218434 TI - Characterization of Escherichia coli NrdH. A glutaredoxin-like protein with a thioredoxin-like activity profile. AB - Ribonucleotides are converted to deoxyribonucleotides by ribonucleotide reductases. Either thioredoxin or glutaredoxin is a required electron donor for class I and II enzymes. Glutaredoxins are reduced by glutathione, thioredoxins by thioredoxin reductase. Recently, a glutaredoxin-like protein, NrdH, was isolated as the functional electron donor for a NrdEF ribonucleotide reductase, a class Ib enzyme, from Lactococcus lactis. The absence of glutathione in this bacterium raised the question of the identity of the intracellular reductant for NrdH. Homologues of NrdH are present in the genomes of Escherichia coli and Salmonella typhimurium, upstream of the genes for the poorly transcribed nrdEF, separated from it by an open reading frame (nrdI) coding for a protein of unknown function. Overexpression of E. coli NrdH protein shows that it is a functional hydrogen donor with higher specificity for the class Ib (NrdEF) than for the class Ia (NrdAB) ribonucleotide reductase. Furthermore, this glutaredoxin-like enzyme is reduced by thioredoxin reductase and not by glutathione. We suggest that several uncharacterized glutaredoxin-like proteins present in the genomes of organisms lacking GSH, including archae, will also react with thioredoxin reductase and be related to the ancestors from which the GSH-dependent glutaredoxins have evolved by the acquisition of a GSH-binding site. We also show that NrdI, encoded by all nrdEF operons, has a stimulatory effect on ribonucleotide reduction. PMID- 9218435 TI - Binding of activated cyclosome to p13(suc1). Use for affinity purification. AB - Previous studies have indicated that a approximately 1,500-kDa complex, designated the cyclosome or anaphase-promoting complex, has a regulated cyclin ubiquitin ligase activity that targets cyclin B for degradation at the end of mitosis. The cyclosome is inactive in the interphase of the embryonic cell cycle and is converted to the active form in late mitosis in a phosphorylation dependent process initiated by protein kinase Cdc2-cyclin B. We show here that the active, phosphorylated form of the cyclosome from clam oocytes binds to p13(suc1), a protein known to associate with Cdc2. The following evidence indicates that the binding of the cyclosome to p13(suc1) is not mediated via the Cdc2-cyclin B complex: (a) activated cyclosome binds to p13(suc1)-Sepharose following its separation from Cdc2-cyclin B by gel filtration chromatography; (b) cyclosome from interphase extracts, activated by a kinase in which cyclin B has been replaced by an N-terminally truncated derivative fused to glutathione S transferase, binds well to p13(suc1)-Sepharose but not to glutathione-agarose. An alternative possibility, that the phosphorylated cyclosome binds directly to a phosphate-binding site of p13(suc1), is supported by the observation that the cyclosome is efficiently eluted from p13(suc1)-Sepharose by phosphate-containing compounds. This information was utilized to develop a procedure for the affinity purification of the cyclosome. A factor abundant in the fraction not adsorbed to p13(suc1)-Sepharose stimulates the activity of purified cyclosome. It is suggested that binding of Suc1 may have a role in the regulation of cyclosome activity. PMID- 9218436 TI - Two distinct TATA-less promoters direct tissue-specific expression of the rat apo B editing catalytic polypeptide 1 gene. AB - The species and tissue specificity of apolipoprotein (apo) B mRNA editing is determined by the expression of apoB editing catalytic polypeptide 1 (APOBEC-1), the cytidine deaminase that catalyzes apoB mRNA editing. To understand the molecular mechanisms that regulate the transcription of APOBEC-1, we characterized rat APOBEC-1 cDNA and genomic DNA. cDNA cloning and RNase protection analysis showed two alternative promoters for the tissue-specific expression of APOBEC-1 in the liver and intestine, Pliv and Pint. Both promoters lack a TATA box, and Pint belongs to the MED-1 class of promoters, which initiate transcription at multiple sites. We also identified two allelic forms of the APOBEC-1 gene from the characterization of two rat APOBEC-1 P1 genomic clones, RE4 and RE5. The RE4 allele is 18 kilobases long and contains six exons and five introns, whereas the RE5 allele contains an additional approximately 8 kilobases of intron sequences and an extra exon encoding a 5'-untranslated region; however, the APOBEC-1 transcripts from the two alleles appear to have similar, if not identical, functions. Transgenic mouse studies showed that Pliv was preferentially used in the liver, kidney, brain, and adipose tissues, whereas Pint was preferentially used in the small intestine, stomach, and lung. Our results suggest that the tissue-specific expression of APOBEC-1 is governed by multiple regulatory elements exerting control over a single coding sequence. The presence or absence of these regulatory elements may determine the tissue specific expression of APOBEC-1 in other mammalian species. PMID- 9218437 TI - Elastin degradation by matrix metalloproteinases. Cleavage site specificity and mechanisms of elastolysis. AB - Insoluble elastin was used as a substrate to characterize the peptide bond specificities of human (HME) and mouse macrophage elastase (MME) and to compare these enzymes with other mammalian metalloproteinases and serine elastases. New amino termini detected by protein sequence analysis in insoluble elastin following proteolytic digestion reveal the P'1 residues in the carboxyl-terminal direction from the scissile bond. The relative proportion of each amino acid in this position reflects the proteolytic preference of the elastolytic enzyme. The predominant amino acids detected by protein sequence analysis following cleavage of insoluble elastin with HME, MME, and 92-kDa gelatinase were Leu, Ile, Ala, Gly, and Val. HME and MME were similar in their substrate specificity and showed a stronger preference for Leu/Ile than did the 92-kDa enzyme. Fibroblast collagenase showed no activity toward elastin. The amino acid residues detected in insoluble elastin following hydrolysis with porcine pancreatic elastase and human neutrophil elastase were predominantly Gly and Ala, with lesser amounts of Val, Phe, Ile, and Leu. There were interesting specificity differences between the two enzymes, however. For both the serine and matrix metalloproteinases, catalysis of peptide bond cleavage in insoluble elastin was characterized by temperature effects and water requirements typical of common enzyme-catalyzed reactions, even those involving soluble substrates. In contrast to what has been observed for collagen, the energy requirements for elastolysis were not extraordinary, consistent with cleavage sites in elastin being readily accessible to enzymatic attack. PMID- 9218438 TI - Synergy between anions and farnesyldiphosphate competitive inhibitors of farnesyl:protein transferase. AB - Investigation of the comparative activities of various inhibitors of farnesyl:protein transferase (FPTase) has led to the observation that the presence of phosphate or pyrophosphate ions in the assay buffer increases the potency of farnesyl diphosphate (FPP) competitive inhibitors. In addition to exploring the phenomenon of phosphate synergy, we report here the effects of various other ions including sulfate, bicarbonate, and chloride on the inhibitory ability of three FPP competitive compounds: Cbz-His-Tyr-Ser(OBn)TrpNH2 (2), Cbz HisTyr(OPO42-)-Ser(OBn)TrpNH2 (3), and alpha-hydroxyfarnesyl phosphonic acid (4). Detailed kinetic analysis of FPTase inhibition revealed a high degree of synergy for compound 2 and each of these ions. Phosphorylation of 2 to give 3 completely eliminated any ionic synergistic effect. Moreover, these ions have an antagonistic effect on the inhibitory potency of compound 4. The anions in the absence of inhibitor exhibit non-competitive inhibition with respect to FPP. These results suggest that phosphate, pyrophosphate, bicarbonate, sulfate, and chloride ions may be binding at the active site of both free enzyme and product bound enzyme with normal substrates. These bound complexes increase the potency of FPP competitive inhibitors and mimic an enzyme:product form of the enzyme. None of the anions studied here proved to be synergistic with respect to inhibition of geranylgeranyl transferase I. These findings provide insight into the mechanism of action of FPP competitive inhibitors for FPTase and point to enzymatic differences between FPTase and geranylgeranyl transferase I that may facilitate the design of more potent and specific inhibitors for these therapeutically relevant target enzymes. PMID- 9218439 TI - cca1 is required for formation of growth-arrested confluent monolayer of rat 3Y1 cells. AB - A novel cDNA fragment, named cca1 (confluent 3Y1 cell-associated 1), was previously isolated on the basis of preferential accumulation of the corresponding mRNA in growth-arrested confluent but not in growing subconfluent rat 3Y1 cells. The cca1 cDNA was found to consist of 5022 nucleotides with an open reading frame of 1905 nucleotides, encoding a protein of 635 amino acids. Unlike the 3Y1 cell case, cca1 mRNA was not detected in confluent 3Y1 BU, 3Y1 BU/pTK, 3Y1-16E6, or F2408 cells, whose growth patterns monitored by phalloidin staining and bromodeoxyuridine incorporation were different from those of the confluent 3Y1 cells. A restoration of the confluent 3Y1-type growth pattern was observed in the cca1 cDNA-introduced 3Y1 BU and 3Y1 BU/pTK cells after reaching confluence but not in the cDNA-introduced 3Y1-16E6 or F2408 cells. The results allow us to conclude that cca1 is required but not sufficient for formation of growth-arrested confluent monolayer of 3Y1 cells. PMID- 9218440 TI - An RNA polymerase III-defective mutation in TATA-binding protein disrupts its interaction with a transcription factor IIIB subunit in drosophila cells. AB - A subunit of the Drosophila RNA polymerase III transcription factor IIIB (TFIIIB) complex has been identified using antibodies directed against the analogous human protein, hIIIB90. This protein has an apparent molecular mass of 105 kDa and has been designated dTAFIII105. Drosophila S-2 cell extracts that were immunodepleted of dTAFIII105 were substantially reduced in their capacity to support tRNA gene transcription. A protein (far Western) blot analysis revealed that dTAFIII105, present in a TFIIIB fraction, directly interacts with TATA-binding protein (TBP). Coimmunoprecipitation assays demonstrated that this protein associates with TBP in S-2 cell extracts. Our previous studies have identified a mutation at position 332 within Drosophila TBP that changes a highly conserved arginine residue to a histidine residue, which renders it specifically defective in its ability to support RNA polymerase III transcription in S-2 cells (Trivedi, A., Vilalta, A., Gopalan, S., and Johnson, D. L. (1996) Mol. Cell. Biol. 16, 6909-6916). We further demonstrate that extracts prepared from a stable cell line expressing epitope-tagged wild-type TBP exhibit an increase in tRNA gene transcription, whereas extracts derived from cells expressing the mutant TBP protein do not. Coimmunoprecipitation assays and far Western blot analysis demonstrate that this mutation in TBP abolishes its ability to stably interact with dTAFIII105. Thus, we have identified both a Drosophila protein that is directly associated with TBP in the TFIIIB complex, dTAFIII105, and an amino acid residue within the highly conserved carboxyl-terminal region of TBP that is critical for dTAFIII105-TBP interactions. PMID- 9218441 TI - Construction of a full-length Ca2+-sensitive adenylyl cyclase/aequorin chimera. AB - Ca2+-sensitive adenylyl cyclases are key integrators of Ca2+ and cAMP signaling. To selectively probe dynamic changes in [Ca2+]i at the plasma membrane where adenylyl cyclases reside, a full-length, Ca2+-inhibitable type VI adenylyl cyclase/aequorin chimera has been constructed by a two-stage polymerase chain reaction method. The expressed adenylyl cyclase/aequorin chimera was appropriately localized to the plasma membrane, as judged by biochemical fractionation and functional analysis. The chimera retained full adenylyl cyclase activity and sensitivity to inhibition by physiological [Ca2+]i elevation. The aequorin portion of the chimeric construct was also capable of measuring changes in [Ca2+] both in vitro and in vivo. When the plasma membrane-tagged aequorin and cytosolic aequorin were compared in their measurement of [Ca2+]i, they showed contrasting sensitivities depending on whether the [Ca2+]i originated from internal stores or capacitative entry. This is the first full-length enzyme aequorin chimera that retains the full biological properties of both aequorin and a Ca2+-sensitive adenylyl cyclase. This novel chimeric Ca2+ sensor provides the unique ability to directly report the dynamics of [Ca2+]i that regulates this Ca2+-sensitive enzyme under a variety of physiological conditions. Since this chimera is localized to the plasma membrane, it can also be used to assess local changes in [Ca2+]i at the plasma membrane as distinct from global changes in [Ca2+]i within the cytosol. PMID- 9218442 TI - Cystine knot of the gonadotropin alpha subunit is critical for intracellular behavior but not for in vitro biological activity. AB - The common alpha subunit of glycoprotein hormones contains five disulfide bonds. Based on the published crystal structure, the assignments are 7-31, 59-87, 10-60, 28-82, and 32-84; the last three comprise the cystine knot, a structure also seen in a variety of growth factors. Previously, we demonstrated that the efficiency of secretion and the ability to form heterodimers by alpha subunits bearing single cysteine residue mutants in the cystine knot were significantly reduced. These results suggested that the cystine knot is critical for the intracellular integrity of the subunit. To assess if the presence of the free thiol affected the secretion kinetics, we constructed paired cysteine mutants of each disulfide bond of the alpha subunit. The secretion rate for these monomers was comparable with wild type except for the alpha-10-60 mutant, which was 40% lower. The recovery of the alpha7-31 and alpha59-87 mutants was greater than 95%, whereas for the cystine knot mutants, it was 20-40%. Co-expression of the wild-type chorionic gonadotropin beta subunit with double cysteine mutants did not enhance the recovery of alpha mutants in the media. Moreover, compared with wild-type, the efficiency of heterodimer formation of the alpha10-60 or alpha32-84 mutants was less than 5%. Because subunit assembly is required for biological activity, studies on the role of these disulfide bonds in signal transduction were not possible. To bypass the assembly step, we exploited the single chain model, where the alpha and beta subunits are genetically fused. The recovery of secreted tethered gonadotropins bearing mutations in the cystine knot was increased significantly. Although dimer-specific monoclonal antibodies discriminated the conformation of single chain alpha10-60 and alpha32-84 mutants from the native heterodimer, these mutants were nevertheless biologically active. Thus, individual bonds of cystine knot are important for secretion and heterodimer formation but not for in vitro bioactivity. Moreover, the data suggest that the native heterodimer configuration is not a prerequisite for receptor binding or signal transduction. PMID- 9218443 TI - Expression of the type II iodothyronine deiodinase in cultured rat astrocytes is selenium-dependent. AB - The iodothyronine deiodinases are a family of selenoproteins that metabolize thyroxine and other thyroid hormones to active and inactive metabolites in a number of tissues including brain. Using primary cultures of rat astroglial cells as a model system, we demonstrate that the mRNA for the type II iodothyronine deiodinase (DII) selenoenzyme is rapidly and markedly induced by forskolin and 8 bromo-cAMP. The induction of DII activity, however, was significantly impaired by culturing cells in selenium-deficient medium for 7 days. Under such conditions, the addition of selenium resulted in a rapid increase in cAMP-induced DII activity that was dose-dependent, with maximal effects noted within 2 h. Cycloheximide blocked this effect of selenium on restoring cAMP-induced DII activity, whereas actinomycin D did not. These data demonstrate that the DII selenoenzyme is expressed in cultured astrocytes and that the induction of DII activity by cAMP analogues appears to be mediated, at least in part, by pretranslational mechanisms. Furthermore, selenium deprivation impairs the expression of DII activity at the level of translation. PMID- 9218444 TI - Catalytic mechanism of the oxidative demethylation of 4-(methoxymethyl)phenol by vanillyl-alcohol oxidase. Evidence for formation of a p-quinone methide intermediate. AB - The catalytic mechanism for the oxidative demethylation of 4 (methoxymethyl)phenol by the covalent flavoprotein vanillyl-alcohol oxidase was studied. Using H218O, it was found that the carbonylic oxygen atom from the product 4-hydroxybenzaldehyde originates from a water molecule. Oxidation of vanillyl alcohol did not result in any incorporation of 18O. Enzyme-monitored turnover experiments revealed that for both substrates a process involving flavin reduction is rate determining. During anaerobic reduction of vanillyl-alcohol oxidase by 4-(methoxymethyl)phenol, a relatively stable spectral intermediate is formed. Deconvolution of its spectral characteristics showed a typical pH independent absorption maximum at 364 nm (epsilon364 nm = 46 mM-1 cm-1). A similar transient species was observed upon anaerobic reduction by vanillyl alcohol. The rate of flavin reduction and synchronous intermediate formation by 4 (methoxymethyl)phenol is 3.3 s-1 and is fast enough to account for turnover (3.1 s-1). The anaerobic decay of the intermediate was too slow (0.01 s-1) to be of catalytical relevance. The reduced binary complex is rapidly reoxidized (1.5 x 10(5) M-1 s-1) and is accompanied with formation and release of product. Oxidation of free-reduced enzyme is an even faster process (3.1 x 10(5) M-1 s-1). The kinetic data for the oxidative demethylation of 4-(methoxymethyl)phenol are in accordance with a ternary complex mechanism in which the reduction rate is rate-limiting. It is proposed that, upon reduction, a binary complex is produced composed of the p-quinone methide of 4-(methoxymethyl)phenol and reduced enzyme. PMID- 9218445 TI - MNN6, a member of the KRE2/MNT1 family, is the gene for mannosylphosphate transfer in Saccharomyces cerevisiae. AB - In yeast Saccharomyces cerevisiae the N-linked sugar chain is modified at different positions by the addition of mannosylphosphate. The mnn6 mutant is deficient in the mannosylphosphate transferase activity toward mannotetraose (Karson, E. M., and Ballou, C. E. (1978) J. Biol. Chem. 253, 6484-6492). We have cloned the MNN6 gene by complementation. It has encoded a 446-amino acid polypeptide with the characteristics of type II membrane protein. The deduced Mnn6p showed a significant similarity to Kre2p/Mnt1p, a Golgi alpha-1, 2 mannosyltransferase involved in O-glycosylation. The null mutant of MNN6 showed a normal cell growth, less binding to Alcian blue, hypersensitivity to Calcoflour White and hygromycin B, and diminished mannosylphosphate transferase activity toward the endoplasmic reticulum core oligosaccharide acceptors (Man8GlcNAc2-PA and Man5GlcNAc2-PA) in vitro, suggesting the involvement of the MNN6 gene in the endoplasmic reticulum core oligosaccharide phosphorylation. However, no differences were observed in N-linked mannoprotein oligosaccharides between Deltaoch1 Deltamnn1 cells and Deltaoch1Deltamnn1Deltamnn6 cells, indicating the existence of redundant genes required for the core oligosaccharide phosphorylation. Based on a dramatic decrease in polymannose outer chain phosphorylation by MNN6 gene disruption and a determination of the mannosylphosphorylation site in the acceptor, it is postulated that the MNN6 gene may be a structural gene encoding a mannosylphosphate transferase, which recognizes any oligosaccharides with at least one alpha-1,2-linked mannobiose unit. PMID- 9218446 TI - Mechanism of quenching of phototransduction. Binding competition between arrestin and transducin for phosphorhodopsin. AB - Quenching of phototransduction in retinal rod cells involves phosphorylation of photoactivated rhodopsin by the enzyme rhodopsin kinase followed by binding of the protein arrestin. Although it has been proposed that the mechanism of arrestin quenching of visual transduction is via steric exclusion of transducin binding to phosphorylated light-activated rhodopsin (P-Rh*), direct evidence for this mechanism is lacking. In this study, we investigated both the role of rhodopsin phosphorylation in modulating its interaction with arrestin and transducin and the proposed binding competition between arrestin and transducin for P-Rh*. While the beta-adrenergic receptor kinase promotes significant arrestin binding to rhodopsin at a phosphorylation stoichiometry of >/=2 mol/mol, rhodopsin kinase promotes arrestin binding at a stoichiometry of approximately 0.9 mol/mol. Moreover, while beta-adrenergic receptor kinase phosphorylation of rhodopsin only modestly decreases transducin binding and activation, rhodopsin kinase phosphorylation of rhodopsin significantly decreases transducin binding and activation. Finally, arrestin competes effectively with transducin for binding to P-Rh* (50% inhibition at approximately 1:1 molar ratio of arrestin:transducin) but has no effect on transducin binding to nonphosphorylated light-activated rhodopsin (Rh*), paralleling the functional inhibition by arrestin on P-Rh*-stimulated transducin activation (50% inhibition at approximately 1.7:1 molar ratio of arrestin:transducin). These results demonstrate that a major role of rhodopsin phosphorylation is to promote high affinity arrestin binding and decrease transducin binding thus allowing arrestin to effectively compete with transducin for binding to photoactivated rhodopsin. PMID- 9218447 TI - 17beta-estradiol potently suppresses cAMP-induced insulin-like growth factor-I gene activation in primary rat osteoblast cultures. AB - Insulin-like growth factor-I (IGF-I) is a key factor in bone remodeling. In osteoblasts, IGF-I synthesis is enhanced by parathyroid hormone and prostaglandin E2 (PGE2) through cAMP-activated protein kinase. In rats, estrogen loss after ovariectomy leads to a rise in serum IGF-I and an increase in bone remodeling, both of which are reversed by estrogen treatment. To examine estrogen-dependent regulation of IGF-I expression at the molecular level, primary fetal rat osteoblasts were co-transfected with the estrogen receptor (hER, to ensure active ER expression), and luciferase reporter plasmids controlled by promoter 1 of the rat IGF-I gene (IGF-I P1), used exclusively in these cells. As reported, 1 microM PGE2 increased IGF-I P1 activity by 5-fold. 17beta-Estradiol alone had no effect, but dose-dependently suppressed the stimulatory effect of PGE2 by up to 90% (ED50 approximately 0.1 nM). This occurred within 3 h, persisted for at least 16 h, required ER, and appeared specific, since 17alpha-estradiol was 100-300-fold less effective. By contrast, 17beta-estradiol stimulated estrogen response element (ERE)-dependent reporter expression by up to 10-fold. 17beta-Estradiol also suppressed an IGF-I P1 construct retaining only minimal promoter sequence required for cAMP-dependent gene activation, but did not affect the 60-fold increase in cAMP induced by PGE2. There is no consensus ERE in rat IGF-I P1, suggesting novel downstream interactions in the cAMP pathway that normally enhances IGF-I expression in skeletal cells. To explore this, nuclear extract from osteoblasts expressing hER were examined by electrophoretic mobility shift assay using the atypical cAMP response element in IGF-I P1. Estrogen alone did not cause DNA-protein binding, while PGE2 induced a characteristic gel shift complex. Co-treatment with both hormones caused a gel shift greatly diminished in intensity, consistent with their combined effects on IGF-I promoter activity. Nonetheless, hER did not bind IGF-I cAMP response element or any adjacent sequences. These results provide new molecular evidence that estrogen may temper the biological effects of hormones acting through cAMP to regulate skeletal IGF-I expression and activity. PMID- 9218448 TI - Membrane potential-generating malate (MleP) and citrate (CitP) transporters of lactic acid bacteria are homologous proteins. Substrate specificity of the 2 hydroxycarboxylate transporter family. AB - Membrane potential generation via malate/lactate exchange catalyzed by the malate carrier (MleP) of Lactococcus lactis, together with the generation of a pH gradient via decarboxylation of malate to lactate in the cytoplasm, is a typical example of a secondary proton motive force-generating system. The mleP gene was cloned, sequenced, and expressed in a malolactic fermentation-deficient L. lactis strain. Functional analysis revealed the same properties as observed in membrane vesicles of a malolactic fermentation-positive strain. MleP belongs to a family of secondary transporters in which the citrate carriers from Leuconostoc mesenteroides (CitP) and Klebsiella pneumoniae (CitS) are found also. CitP, but not CitS, is also involved in membrane potential generation via electrogenic citrate/lactate exchange. MleP, CitP, and CitS were analyzed for their substrate specificity. The 2-hydroxycarboxylate motif R1R2COHCOOH, common to the physiological substrates, was found to be essential for transport although some 2 oxocarboxylates could be transported to a lesser extent. Clear differences in substrate specificity among the transporters were observed because of different tolerances toward the R substituents at the C2 atom. Both MleP and CitP transport a broad range of 2-hydroxycarboxylates with R substituents ranging in size from two hydrogen atoms (glycolate) to acetyl and methyl groups (citromalate) for MleP and two acetyl groups (citrate) for CitP. CitS was much less tolerant and transported only citrate and at a low rate citromalate. The substrate specificities are discussed in the context of the physiological function of the transporters. PMID- 9218449 TI - Cell type-specific inhibition of keratinocyte collagenase-1 expression by basic fibroblast growth factor and keratinocyte growth factor. A common receptor pathway. AB - Collagenase-1 is invariantly expressed by migrating basal keratinocytes in all forms of human skin wounds, and its expression is induced by contact with native type I collagen. However, net differences in enzyme production between acute and chronic wounds may be modulated by soluble factors present within the tissue environment. Basic fibroblast growth factor (bFGF, FGF-2) and keratinocyte growth factor (KGF, FGF-9), which are produced during wound healing, inhibited collagenase-1 expression by keratinocytes in a dose-dependent manner. However, KGF was >100-fold more effective than bFGF at inhibiting collagenase-1 expression, suggesting that this differential signaling is transduced via an FGF receptor that binds these ligands with different affinities. Reverse transcriptase-polymerase chain reaction analysis of human keratinocyte mRNA for fibroblast growth factor receptors (FGFRs) revealed expression of only FGFR-2 IIIb, the KGF-specific receptor, which also binds bFGF with low affinity, and FGFR-3 IIIb, which does not bind bFGF or KGF. FGFRs that bind bFGF with high affinity were not detected. Our results suggest that bFGF and KGF inhibit collagenase-1 expression through the KGF cell-surface receptor (FGFR-2 IIIb). Because bFGF induces collagenase-1 in most cell types, cell-specific expression of FGFR family members may dictate the regulation of matrix metalloproteinases in a tissue-specific manner. PMID- 9218450 TI - ATP-, K+-dependent heptamer exchange reaction produces hybrids between GroEL and chaperonin from Thermus thermophilus. AB - Chaperonin from Thermus thermophilus (Tcpn6014.Tcpn107) splits at the plane between two Tcpn607 rings into two parts in a solution containing ATP and K+ (Ishii, N., Taguchi, H., Sasabe, H., and Yoshida, M. (1995) FEBS Lett. 362, 121 125). When Escherichia coli GroEL14 was additionally included in the solution described above, hybrid chaperonins GroEL7.Tcpn607 and GroEL7. Tcpn607.Tcpn107 were formed rapidly (<20 s) at 37 degrees C. The hybrid was also formed from Tcpn6014 and GroEL14 but not from a mutant GroEL14 lacking ATPase activity. The hybrid formation was saturated at approximately 300 microM ATP and approximately 300 mM K+. These results imply that GroEL14 also splits and undergoes a heptamer exchange reaction with Thermus chaperonin under nearly physiological conditions. Similar to parent chaperonins, the isolated hybrid chaperonins exhibited ATPase activity that was susceptible to inhibition by Tcpn107 or GroES7 and mediated folding of other proteins. Once formed, the hybrid chaperonins were stable, and the parent chaperonins were not regenerated from the isolated hybrids under the same conditions in which the hybrids had been formed. Only under conditions in which GroEL in the hybrids was selectively destroyed, such as incubation at 70 degrees C, Thermus chaperonin, but not GroEL14, was regenerated from the hybrid. Therefore, the split reaction may not be an obligatory event repeated in each turnover of the chaperonin functional cycles but an event that occurs only when chaperonin is first exposed to ATP/K+. PMID- 9218451 TI - Binding of different divalent cations to the active site of avian sarcoma virus integrase and their effects on enzymatic activity. AB - Retroviral integrases (INs) contain two known metal binding domains. The N terminal domain includes a zinc finger motif and has been shown to bind Zn2+, whereas the central catalytic core domain includes a triad of acidic amino acids that bind Mn2+ or Mg2+, the metal cofactors required for enzymatic activity. The integration reaction occurs in two distinct steps; the first is a specific endonucleolytic cleavage step called "processing," and the second is a polynucleotide transfer or "joining" step. Our previous results showed that the metal preference for in vitro activity of avian sarcoma virus IN is Mn2+ > Mg2+ and that a single cation of either metal is coordinated by two of the three critical active site residues (Asp-64 and Asp-121) in crystals of the isolated catalytic domain. Here, we report that Ca2+, Zn2+, and Cd2+ can also bind in the active site of the catalytic domain. Furthermore, two zinc and cadmium cations are bound at the active site, with all three residues of the active site triad (Asp-64, Asp-121, and Glu-157) contributing to their coordination. These results are consistent with a two-metal mechanism for catalysis by retroviral integrases. We also show that Zn2+ can serve as a cofactor for the endonucleolytic reactions catalyzed by either the full-length protein, a derivative lacking the N-terminal domain, or the isolated catalytic domain of avian sarcoma virus IN. However, polynucleotidyl transferase activities are severely impaired or undetectable in the presence of Zn2+. Thus, although the processing and joining steps of integrase employ a similar mechanism and the same active site triad, they can be clearly distinguished by their metal preferences. PMID- 9218452 TI - Characterization of PKIgamma, a novel isoform of the protein kinase inhibitor of cAMP-dependent protein kinase. AB - Attempts to understand the physiological roles of the protein kinase inhibitor (PKI) proteins have been hampered by a lack of knowledge concerning the molecular heterogeneity of the PKI family. The PKIgamma cDNA sequence determined here predicted an open reading frame of 75 amino acids, showing 35% identity to PKIalpha and 30% identity to PKIbeta1. Residues important for the high affinity of PKIalpha and PKIbeta1 as well as nuclear export of the catalytic (C) subunit of cAMP-dependent protein kinase were found to be conserved in PKIgamma. Northern blot analysis showed that a 1.3-kilobase PKIgamma message is widely expressed, with highest levels in heart, skeletal muscle, and testis. RNase protection analysis revealed that in most tissues examined PKIgamma is expressed at levels equal to or higher than the other known PKI isoforms and that in several mouse derived cell lines, PKIgamma is the predominant PKI message. Partial purification of PKI activities from mouse heart by DEAE ion exchange chromatography resolved two major inhibitory peaks, and isoform-specific polyclonal antibodies raised against recombinant PKIalpha and PKIgamma identified these inhibitory activities to be PKIalpha and PKIgamma. A comparison of inhibitory potencies of PKIalpha and PKIgamma expressed in Escherichia coli revealed that PKIgamma was a potent competitive inhibitor of Calpha phosphotransferase activity in vitro (Ki = 0.44 nM) but is 6-fold less potent than PKIalpha (Ki = 0.073 nM). Like PKIalpha, PKIgamma was capable of blocking the nuclear accumulation of Flag-tagged C subunit in transiently transfected mammalian cells. Finally, the murine PKIgamma gene was found to overlap the murine adenosine deaminase gene on mouse chromosome 2. These results demonstrate that PKIgamma is a novel, functional PKI isoform that accounts for the previously observed discrepancy between PKI activity and PKI mRNA levels in several mammalian tissues. PMID- 9218453 TI - Ligand-induced internalization of cholecystokinin receptors. Demonstration of the importance of the carboxyl terminus for ligand-induced internalization of the rat cholecystokinin type B receptor but not the type A receptor. AB - Internalization of a variety of different heptahelical G protein-coupled receptors has been shown to be influenced by a number of different structural determinants of the receptors, including the carboxyl terminus. To investigate the role of the carboxyl terminus of cholecystokinin (CCK) receptors in receptor internalization, the rat wild type (WT) CCK-A receptor (WT CCKAR) and the rat WT CCK-B receptor (WT CCKBR) were truncated after amino acid residue 399 (CCKAR Tr399) and 408 (CCKBR Tr408), thereby deleting the carboxyl-terminal 45 and 44 residues, respectively. All WT and mutant CCK receptors were stably expressed in NIH/3T3 cells. Internalization of the CCKAR Tr399 was not significantly different from the WT CCKAR. In contrast, internalization of the CCKBR Tr408 was decreased to 26% compared with the WT CCKBR internalization of 92%. The mutation of all 10 serine and threonine residues (as potential phosphorylation sites) in the carboxyl terminus of the CCKBR to alanines (mutant CCKBR DeltaS/T) could account for the majority of this effect (39% internalization). All mutant receptors displayed similar ligand binding characteristics, G protein coupling, and signal transduction as their respective WT receptors, indicating that the carboxyl termini are not necessary for these processes. Thus, internalization of the CCKBR, unlike that of the CCKAR, depends on the carboxyl terminus of the receptor. These results suggest that, despite the high degree of homology between CCKAR and CCKBR, the structural determinants that mediate the interaction with the endocytic pathway reside in different regions of the receptors. PMID- 9218454 TI - Renin-1 is essential for normal renal juxtaglomerular cell granulation and macula densa morphology. AB - The secretion of renin from granules stored in renal juxtaglomerular cells plays a key role in blood pressure homeostasis. The synthesis and release of renin and the extent of granulation is regulated by several mechanisms including signaling from the macula densa, neuronal input, and blood pressure. Through the use of a gene-targeting vector containing homology arms generated using the polymerase chain reaction, we have inactivated the Ren-1(d) gene, one of two mouse genes encoding renin, and report that lack of renin-1(d) results in altered morphology of the macula densa of the kidney distal tubule and complete absence of juxtaglomerular cell granulation. Furthermore, Ren-1(d-/-) mice exhibit sexually dimorphic hypotension. The altered growth morphology of the macula densa in Ren 1(d)-null mice should provide a tool for the investigation of the JG cell-macula densa signaling. Furthermore, the current data indicate that expression of the Ren-1(d) gene is a prerequisite for the formation of storage granules, even though the related protein renin-2 is present in these mice, suggesting that renin-1(d) and renin-2 are secreted by distinct pathways in vivo. PMID- 9218455 TI - The influence of antisense oligonucleotide-induced RNA structure on Escherichia coli RNase H1 activity. AB - The ability of Escherichia coli RNase H1 to hydrolyze structured substrates containing antisense oligonucleotides preannealed to a 47-mer RNA was compared with its ability to hydrolyze unstructured substrates containing antisense oligonucleotides duplexed with 13-mer RNA. These results demonstrate that when antisense oligonucleotides were bound to structured RNA, the resultant duplexes were cleaved at rates significantly slower than when the same oligonucleotides were bound to unstructured oligoribonucleotides. Structured substrates exhibited fewer cleavage sites, and each cleavage site was cleaved less rapidly than in unstructured substrates. Furthermore, the enzymatic activity of E. coli RNase H1 for the structured substrates was most affected when the cleavage sites corresponding to the enzymatically most active sites on the unstructured substrates were blocked in the structured substrates. Molecular modeling suggests that the observed ablation of RNase H activity was due to the steric hindrance of the enzyme by the structured RNA, i.e. steric interference of the phosphate groups on the substrate and/or the binding site of the enzyme. When chimeric oligonucleotides composed of a five-base deoxynucleotide sequence flanked by chemically modified nucleotides were bound to structured RNA, the resultant duplexes were even worse substrates for RNase H. These results offer further insights into the role of antisense-induced RNA structure on RNase H activity and may facilitate the design of effective antisense oligonucleotides. PMID- 9218456 TI - Syk is required for BCR-mediated activation of p90Rsk, but not p70S6k, via a mitogen-activated protein kinase-independent pathway in B cells. AB - The tyrosine kinases Syk and Lyn are activated in B lymphocytes following antibody induced cross-linking of the B cell receptor for antigen (BCR). It has been suggested that activation of Syk is dependent on Lyn. We tested this hypothesis by comparing the phosphorylation and activation of several downstream effector molecules in parental DT40, DT40Syk- and DT40Lyn- B cells. The phosphorylation and activation of p90Rsk was ablated in Syk-deficient B cells but unaffected in Lyn-deficient B cells while the phosphorylation/activation of Ras GTPase activating protein (Ras GAP) and mitogen activated protein (MAP) kinase required both Syk and Lyn. Thus, these data indicate that Syk can be activated in the absence of Lyn after BCR cross-linking and results in the activation of p90Rsk via a MAP kinase-independent pathway in DT40Lyn- cells. We also demonstrated that BCR mediates the activation of p70S6k. However, activation of p70S6k in DT40Syk- and DT40Lyn- cells was comparable with that observed in parental cells. Thus, either Syk or Lyn may be sufficient for activation of p70S6k, or activation of p70S6k occurs independently of both Syk and Lyn. The kinase activity of Syk was required for the phosphorylation/activation of each of these downstream effector molecules but only the phosphorylation of Ras GAP was affected in cells expressing a mutant of Syk in which tyrosines 525 and 526 were substituted to phenlyalanines. PMID- 9218457 TI - Structure of the murine CD156 gene, characterization of its promoter, and chromosomal location. AB - The murine cell surface antigen mCD156 is a glycoprotein that is expressed in monocytic cell lines and consists of a metalloprotease domain, a disintegrin domain, a cysteine-rich domain, and an epidermal growth factor-like domain in the extracellular region. The mCD156 gene is composed of 24 exons and 23 introns and spans approximately 14 kilobases. The first exon encodes most of the signal peptide sequence, and the transmembrane region is encoded by a single exon (19). In contrast, the other regions are composed of multiple exons. Of these, exons 7 12 and 12-15 encode a metalloprotease domain and a disintegrin domain, respectively. Sequence analysis of the 5'-flanking DNA revealed many potential regulatory motifs. Chloramphenicol acetyltransferase analysis demonstrated that nucleotides at positions -183, -334, and -623 contained cis-acting enhancing elements in a mouse monocytic cell line, aHINS-B3. Nucleotides at positions -183 and -390 contained elements responsible for lipopolysaccharide (LPS) inducibility, although several other 5'-flanking regions were also involved in LPS responsiveness. Regions -202, -507, and -659 play a role in interferon-gamma inducibility. Some of the potential regulatory motifs and other unknown cis elements may be involved in the constitutive expression, and LPS and interferon gamma inducibilities. The mCD156 gene was mapped to chromosome 7, region F3-F4. PMID- 9218458 TI - Structure of cardiac muscle troponin C unexpectedly reveals a closed regulatory domain. AB - The regulation of cardiac muscle contraction must differ from that of skeletal muscles to effect different physiological and contractile properties. Cardiac troponin C (TnC), the key regulator of cardiac muscle contraction, possesses different functional and Ca2+-binding properties compared with skeletal TnC and features a Ca2+-binding site I, which is naturally inactive. The structure of cardiac TnC in the Ca2+-saturated state has been determined by nuclear magnetic resonance spectroscopy. The regulatory domain exists in a "closed" conformation even in the Ca2+-bound (the "on") state, in contrast to all predicted models and differing significantly from the calcium-induced structure observed in skeletal TnC. This structure in the Ca2+-bound state, and its subsequent interaction with troponin I (TnI), are crucial in determining the specific regulatory mechanism for cardiac muscle contraction. Further, it will allow for an understanding of the action of calcium-sensitizing drugs, which bind to cardiac TnC and are known to enhance the ability of cardiac TnC to activate cardiac muscle contraction. PMID- 9218459 TI - Organization and myogenic restricted expression of the murine serum response factor gene. A role for autoregulation. AB - Serum response factor (SRF), a member of an ancient family of DNA-binding proteins, is generally assumed to be a ubiquitous transcription factor involved in regulating growth factor-responsive genes. However, avian SRF was recently shown (Croissant, J. D., Kim, J.-H., Eichele, G., Goering, L., Lough, J., Prywes, R., and Schwartz, R. J. (1996) Dev. Biol. 177, 250-264) to be preferentially expressed in myogenic lineages and is required for regulating post-replicative muscle gene expression. Given the central importance of SRF for the muscle tissue restricted expression of the striated alpha-actin gene family, we wanted to determine how SRF might contribute to this muscle-restricted expression. Here we have characterized the murine SRF genomic locus, which has seven exons interrupted by six introns, with the entire locus spanning 11 kilobases. Murine SRF transcripts were processed to two 3'-untranslated region polyadenylation signals, yielding 4.5- and 2.5-kilobase mRNA species. Murine SRF mRNA levels were the highest in adult skeletal and cardiac muscle, but barely detected in liver, lung, and spleen tissues. During early mouse development, in situ hybridization analysis revealed enrichment of SRF transcripts in the myotomal portion of somites, the myocardium of the heart, and the smooth muscle media of vessels of mouse embryos. Likewise, murine SRF promoter activity was tissue-restricted, being 80-fold greater in primary skeletal myoblasts than in liver-derived HepG2 cells. In addition, SRF promoter activity increased 6-fold when myoblasts withdrew from the cell cycle and fused into differentiated myotubes. A 310-base pair promoter fragment depended upon multiple intact serum response elements in combination with Sp1 sites for maximal myogenic restricted activity. Furthermore, cotransfected SRF expression vector stimulated SRF promoter transcription, whereas dominant-negative SRF mutants blocked SRF promoter activity, demonstrating a positive role for an SRF-dependent autoregulatory loop. PMID- 9218460 TI - Calumenin, a Ca2+-binding protein retained in the endoplasmic reticulum with a novel carboxyl-terminal sequence, HDEF. AB - We have identified and characterized a cDNA encoding a novel Ca2+-binding protein named calumenin from mouse heart by the signal sequence trap method. The deduced amino acid sequence (315 residues) of calumenin contains an amino-terminal signal sequence and six Ca2+-binding (EF-hand) motifs and shows homology with reticulocalbin, Erc-55, and Cab45. These proteins seem to form a new subset of the EF-hand protein family expressed in the lumen of the endoplasmic reticulum (ER) and Golgi apparatus. Purified calumenin had Ca2+-binding ability. The carboxyl-terminal tetrapeptide His-Asp-Glu-Phe was shown to be responsible for retention of calumenin in ER by the retention assay, immunostaining with a confocal laser microscope, and the deglycosylation assay. This is the first report indicating that the Phe residue is included in the ER retention signal. Calumenin is expressed most strongly in heart of adult and 18.5-day embryos. The calumenin gene (Calu) was mapped at the proximal portion of mouse chromosome 7. PMID- 9218461 TI - Molecular cloning of human plasma membrane phospholipid scramblase. A protein mediating transbilayer movement of plasma membrane phospholipids. AB - The rapid movement of phospholipids (PL) between plasma membrane leaflets in response to increased intracellular Ca2+ is thought to play a key role in expression of platelet procoagulant activity and in clearance of injured or apoptotic cells. We recently reported isolation of a approximately 37-kDa protein in erythrocyte membrane that mediates Ca2+-dependent movement of PL between membrane leaflets, similar to that observed upon elevation of Ca2+ in the cytosol (Basse, F., Stout, J. G., Sims, P. J., and Wiedmer, T. (1996) J. Biol. Chem. 271, 17205-17210). Based on internal peptide sequence obtained from this protein, a 1,445-base pair cDNA was cloned from a K-562 cDNA library. The deduced "PL scramblase" protein is a proline-rich, type II plasma membrane protein with a single transmembrane segment near the C terminus. Antibody against the deduced C terminal peptide was found to precipitate the approximately 37-kDa red blood cell protein and absorb PL scramblase activity, confirming the identity of the cloned cDNA to erythrocyte PL scramblase. Ca2+-dependent PL scramblase activity was also demonstrated in recombinant protein expressed from plasmid containing the cDNA. Quantitative immunoblotting revealed an approximately 10-fold higher abundance of PL scramblase in platelet ( approximately 10(4) molecules/cell) than in erythrocyte ( approximately 10(3) molecules/cell), consistent with apparent increased PL scramblase activity of the platelet plasma membrane. PL scramblase mRNA was found in a variety of hematologic and nonhematologic cells and tissues, suggesting that this protein functions in all cells. PMID- 9218462 TI - Characterization of the rat thyroid iodide transporter using anti-peptide antibodies. Relationship between its expression and activity. AB - Anti-peptide antibodies directed against the C-terminal portion (amino acids 603 618) of the rat thyroid iodide transporter (rTIT) have been produced to characterize the molecular forms of rTIT in the rat thyroid and in the functional rat thyroid cell line, FRTL-5. rTIT is located on the basolateral membrane of rat thyroid follicular cells and randomly distributed on the plasma membrane of FRTL 5 cells that do not exhibit cell polarity. The major rTIT component corresponds to an 80-90-kDa glycosylated protein. After treatment of cell membrane fractions with N-glycosidase F or incubation of FRTL-5 cells with tunicamycin, rTIT has an apparent molecular mass of about 55 kDa. FRTL-5 cells cultured in the presence of TSH exhibit a high rTIT content and a high iodide uptake activity (IUA). Upon either removal of TSH or addition of cycloheximide, IUA declines more rapidly than rTIT. The half-life of rTIT was about 4 days. Re-exposure of 7-day TSH deprived FRTL-5 cells to TSH causes a rapid synthesis of the glycosylated rTIT but a delayed re-induction of IUA. Tunicamycin totally prevents the TSH-dependent re-expression and activity of rTIT. Our data bring basic information on the location, structure, and turnover of rTIT and suggest that its activity is subjected to diverse control mechanisms including regulatory proteins. PMID- 9218463 TI - Functional interactions between the estrogen receptor and the transcription activator Sp1 regulate the estrogen-dependent transcriptional activity of the vitellogenin A1 io promoter. AB - Two distinct, TATA box-containing promoters regulate the transcriptional activity of the Xenopus vitellogenin A1 gene. These two promoters are of different strength and are separated by 1.8 kilobase pairs of untranslated sequence. Estrogen receptor (ER) and its ligand, 17beta-estradiol, induce the activity of both promoters. The estrogen response elements (EREs) are located proximal to the downstream i promoter while no ERE-like sequences have been identified in the vicinity of the upstream io promoter. We show here, that transcriptional activity of the upstream io promoter is Sp1-dependent. Moreover, we demonstrate that estrogen inducibility of the io promoter results from functional interactions between the io bound Sp1 and the ER bound at the proximity of i. Functional interactions between Sp1 and ER do not require the presence of a TATA box for transcriptional activation, as is demonstrated using the acyl-CoA oxidase promoter. The relative positions that ER and Sp1 occupy with respect to the initiation site determines whether these two transcription activators can synergize for transcription initiation. PMID- 9218464 TI - Specific activation of a c-Jun NH2-terminal kinase isoform and induction of neurite outgrowth in PC-12 cells by staurosporine. AB - Staurosporine, a protein kinase inhibitor, is known to mimic the effect of nerve growth factor (NGF) in promoting neurite outgrowth. To elucidate the mechanism by which staurosporine induces neurite outgrowth in PC-12 cells, we performed an in gel kinase assay using myelin basic protein as a substrate, and found that staurosporine induced the activation of a kinase with an apparent molecular mass of 57 kDa. The dose of staurosporine required to activate this kinase was consistent with that required to induce neurite outgrowth. Interestingly, the staurosporine-activated kinase was immunoprecipitated by anti-c-Jun NH2-terminal kinase (JNK) isoforms antibody, but not by anti-JNK1-specific antibody or anti ERK1 antibody, raising the possibility that this kinase is a novel JNK isoform. The substrate specificity of the kinase was distinct from those of osmotic shock activated JNKs and NGF-activated ERK1. The kinase phosphorylates transcription factors including c-Jun, Elk-1, and ATF2, as well as myelin basic protein, suggesting that it plays a role in gene induction. Furthermore, staurosporine induced immediate-early genes including Nur77 and fos, but not jun. The activation of the staurosporine-activated kinase, as well as the induction of neurite outgrowth, did not require Ras function, while Ras was required for the activation of ERKs and neurite outgrowth induced by NGF. Taken together, these results indicate staurosporine specifically activates a JNK isoform, which may contribute to biological activities including neurite outgrowth. PMID- 9218465 TI - Mutational analysis reveals that all-trans-retinoic acid, 9-cis-retinoic acid, and antagonist interact with distinct binding determinants of RARalpha. AB - Retinoids exert their pleiotropic effects on cell differentiation and proliferation through specific nuclear receptors, the retinoic acid receptors (RARs) and retinoid X receptors (RXRs). Two biologically highly active natural retinoids have been identified, all-trans-retinoic acid (t-RA) and 9-cis-retinoic acid (9-cis-RA). The RXRs exclusively bind 9-cis-RA, whereas the RARs bind both isomers of RA with comparable affinity. Recently published results suggest that RARs have the same binding site for t-RA and 9-cis-RA but with different determinants (1-3). Antagonist binding on RARalpha has been suggested to induce distinct conformational changes in comparison with agonist binding. To elucidate the region minimally required for efficient binding of agonist (t-RA and 9-cis RA) and antagonist Ro 41-5253 to the RARalpha, we generated N- and C-terminally truncated mutants of the receptor. Characterization of these deletion mutant proteins using protease mapping and ligand binding experiments revealed that different parts of the ligand-binding domain are necessary for t-RA, 9-cis-RA, and antagonist binding. Three distinct regions of the ligand-binding domain of the human retinoic acid receptor-alpha are required for binding of t-RA (RARalpha187-402), 9-cis-RA (RARalpha188-409), and the antagonist Ro 41-5253 (RARalpha226-414). PMID- 9218466 TI - Regulation of G protein-coupled receptor kinases by calmodulin and localization of the calmodulin binding domain. AB - G protein-coupled receptor kinases (GRKs) specifically phosphorylate and regulate the activated form of multiple G protein-coupled receptors. Recent studies have revealed that GRKs are also subject to regulation. In this regard, GRK2 and GRK5 can be phosphorylated and either activated or inhibited, respectively, by protein kinase C. Here we demonstrate that calmodulin, another mediator of calcium signaling, is a potent inhibitor of GRK activity with a selectivity for GRK5 (IC50 approximately 50 nM) > GRK6 >> GRK2 (IC50 approximately 2 microM) >> GRK1. Calmodulin inhibition of GRK5 is mediated via a reduced ability of the kinase to bind to both receptor and phospholipid. Interestingly, calmodulin also activates autophosphorylation of GRK5 at sites distinct from the two major autophosphorylation sites on GRK5. Moreover, calmodulin-stimulated autophosphorylation directly inhibits GRK5 interaction with receptor even in the absence of calmodulin. Using glutathione S-transferase-GRK5 fusion proteins either to inhibit calmodulin-stimulated autophosphorylation or to bind directly to calmodulin, we determined that an amino-terminal domain of GRK5 (amino acids 20-39) is sufficient for calmodulin binding. This domain is abundant in basic and hydrophobic residues, characteristics typical of calmodulin binding sites, and is highly conserved in GRK4, GRK5, and GRK6. These studies suggest that calmodulin may serve a general role in mediating calcium-dependent regulation of GRK activity. PMID- 9218467 TI - Interactions of nucleotide release factor Dss4p with Sec4p in the post-Golgi secretory pathway of yeast. AB - SEC4 is an essential gene encoding a small GTPase that is involved in Golgi to cell surface transport in Saccharomyces cerevisiae and is a paradigm for studies on the mode of action of Rab proteins. We describe here the features of interaction of Sec4p with the accessory protein Dss4p. Dss4p is found both on membranes and in the cytosol; however, it is the membrane fraction that is complexed to Sec4p. Dss4p, like its mammalian counterpart, Mss4, binds zinc, and disruption of the zinc-binding site disrupts the ability of the protein to interact with Sec4p. DSS4 overexpression can rescue the lethal phenotype of two alleles of SEC4, corresponding to dominant mutations of Ras. We demonstrate that this suppression is due to the ability of Dss4p to form a tight complex with the mutant forms of Sec4p and hence sequester the mutant protein from its inhibitory effect. These results imply an in vivo role for Dss4p as a guanine nucleotide dissociation stimulator. In vitro the protein has the ability to stimulate the dissociation rate of both GDP and GTP from Sec4p. We examined the relationship of GDI1 and DSS4 with SEC4 both genetically and biochemically. These results exclude a role for DSS4 in the recruitment of Sec4p/GDI onto membranes. PMID- 9218468 TI - Reconstitution of bile acid transport in a heterologous cell by cotransfection of transporters for bile acid uptake and efflux. AB - The rat liver canalicular bile acid transporter/ecto-ATPase/cell CAM 105 (CBATP) is a 110-kDa transmembrane phosphoglycoprotein that is thought to have bile acid efflux, ecto-ATPase, and cell adhesion properties. Its extracellular amino terminal domain is highly homologous to carcinoembryonic antigen (CEA), a glycophosphatidyl inositol-anchored membrane protein with cell adhesion properties and a marker for adenocarcinoma. In the current study, we examined the possibility of more clearly defining the role of CBATP in bile acid efflux by cotransfecting a heterologous cell, the COS cell, with cDNAs for a bile acid importer, the ileal bile acid transporter (IBAT), as well as for CBATP. The results show that when IBAT mediates uptake of [3H]taurocholate to a level 20 fold higher than that achieved previously by nonspecific pinocytosis, CBATP mediates time-, temperature- and concentration-dependent efflux. Efflux of [3H]taurocholate mediated by CBATP in the cotransfected COS cells is saturable and has curvilinear kinetic characteristics (Vmax = 400 pmol/mg protein/min, Km = 70 microM). It is inhibited by 4,4'-diisothiocyanostilbene-2,2-disulfonic acid and dependent on ATP but not dependent on membrane potential. Although CEA could not mediate bile acid efflux in COS cells cotransfected with IBAT and CEA, efflux of [3H]taurocholate was detected in COS cells cotransfected with IBAT and a chimeric molecule having the carboxyl-terminal tail and membrane spanning domain of CBATP and the amino-terminal extracellular tail of CEA. Taken together, these data provide further evidence that CBATP confers bile acid efflux properties on heterologous cells and that its cytoplasmic tail and membrane spanning segment are integral to this property. The data also establish a model system for more clearly defining the molecular determinants of bile acid transport mediated by this molecule. PMID- 9218469 TI - Characterization of the nucleoside triphosphate phosphohydrolase and helicase activities of the reovirus lambda1 protein. AB - Previous studies have shown that the reovirus lambda1 core protein harbors a putative nucleotide-binding motif and exhibits an affinity for nucleic acids. In addition, a nucleoside triphosphate phosphohydrolase activity present in reovirus cores has been recently assigned to lambda1 using gene reassortment analysis. In this study, it was demonstrated that the recombinant lambda1 protein, expressed in the yeast Pichia pastoris, is able to hydrolyze nucleoside 5'-triphosphates or deoxynucleoside 5'-triphosphates. This activity was absolutely dependent on the presence of a divalent cation, Mg2+ or Mn2+. The protein can also unwind double stranded nucleic acid molecules in the presence of a nucleoside 5'-triphosphate or deoxynucleoside 5'-triphosphate. These results provide the first biochemical evidence that the reovirus lambda1 protein is a nucleoside triphosphate phosphohydrolase/helicase and strongly support the idea that lambda1 participates in transcription of the viral genome. PMID- 9218470 TI - Ligand-independent dimerization of the extracellular domain of the leptin receptor and determination of the stoichiometry of leptin binding. AB - The leptin receptor is a class I transmembrane protein with either a short or a long cytoplasmic domain. Using chemical cross-linking we have analyzed the binding of leptin to its receptor. Cross-linking of radiolabeled leptin to different isoforms of the leptin receptor expressed on COS-1 cells reveals leptin receptor monomer, homodimer, and oligomer complexes. Cotransfection of the long and short form of the leptin receptor did not provide any evidence for the formation of heterodimer complexes. Soluble forms consisting of either the entire extracellular domain or the two cytokine receptor homologous domains of the leptin receptor were purified to homogeneity from recombinant baculovirus infected insect cells by leptin affinity chromatography. Gel filtration chromatography showed that these proteins exist in a dimeric form. Analysis of the complex formed between soluble leptin receptor and leptin by native polyacrylamide gel electrophoresis, and data obtained from the amino acid composition of the complex provide direct evidence that the extracellular domain of the leptin receptor binds leptin in a 1:1 ratio. PMID- 9218471 TI - Recombinant expression of the MAL proteolipid, a component of glycolipid-enriched membrane microdomains, induces the formation of vesicular structures in insect cells. AB - The MAL proteolipid has been identified as a component of glycolipid-enriched membrane microdomains resistant to detergent solubilization in epithelial Madin Darby canine cells, as well as in T lymphocytes and in myelin-forming cells. To study the function of the MAL proteolipid we have ectopically expressed a tagged form of MAL in both mammalian and insect cellular backgrounds. Immunofluorescence analysis in transiently transfected COS-7 cells showed the presence of MAL in large vesicular structures, and biochemical analysis identified MAL in the fraction of membranes resistant to Triton X-100 solubilization. Electron microscopic analysis showed that the expression of MAL in Sf21 cells morphologically resulted in the intracellular accumulation of large vesicles with a diameter from 200 to greater than 700 nm that were absent in uninfected or control infected cultures. Thus, ectopic expression of MAL in this heterologous expression system was sufficient to drive the formation of vesicles with a size similar to that of the vesicles detected in mammalian cells. These vesicles were clearly different from the caveolae-like vesicles induced by caveolin expression, as evidenced by co-infection experiments using a recombinant caveolin baculovirus. Taken together, these results suggest that the MAL proteolipid might play a role as a component of the machinery of vesiculation of glycolipid enriched membranes. PMID- 9218472 TI - Binding of upstream stimulatory factor and a cell-specific activator to the calcitonin/calcitonin gene-related peptide enhancer. AB - The calcitonin/calcitonin gene-related peptide (CT/CGRP) gene is selectively transcribed in thyroid C cells and neurons. We have previously shown that the rat CT/CGRP cell-specific enhancer is synergistically regulated by a helix-loop-helix (HLH) protein and the OB2 octamer-binding protein. In this report, we show that the HLH-OB2 enhancer is required for full promoter activity, even in the context of other HLH elements. Since this enhancer appears to be a major controlling element, we have characterized the HLH and OB2 DNA binding proteins. We have identified the major HLH complex as a heterodimer of the ubiquitous upstream stimulatory factor (USF)-1 and USF-2 proteins. USF bound the enhancer with a reasonably high affinity (KD 1.6 nM), comparable to other genes. Characterization of a series of mutations revealed that a portion of the HLH motif is also recognized by OB2 and confirmed that HLH activity requires OB2. We have shown that OB2 is a single DNA binding protein based on UV cross-linking studies. The 68-kDa protein-DNA complex was detected only in C cell lines, including a human C cell line that has robust HLH-OB2 enhancer activity. These results suggest that the calcitonin/CGRP gene is controlled by the combinatorial activity of a ubiquitous USF HLH heterodimer and an associated cell-specific activator. PMID- 9218473 TI - Mapping the ends of transmembrane segments in a polytopic membrane protein. Scanning N-glycosylation mutagenesis of extracytosolic loops in the anion exchanger, band 3. AB - Band 3, the anion exchanger of human erythrocytes, contains up to 14 transmembrane (TM) segments and has a single endogenous site of N-glycosylation at Asn642 in extracellular (EC) loop 4. The requirements for N-glycosylation of EC loops and the topology of this polytopic membrane protein were determined by scanning N-glycosylation mutagenesis and cell-free translation in a reticulocyte lysate supplemented with microsomal membranes. The endogenous and novel acceptor sites located near the middle of the 35 residue EC loop 4 were efficiently N glycosylated; however, no N-glycosylation occurred at sites located within sharply defined regions close to the adjacent TM segments. Acceptor sites located in the center of EC loop 3, which contains 25 residues, were poorly N glycosylated. Expansion of this loop with a 4-residue insert containing an acceptor site increased N-glycosylation. Acceptor sites located in short (<10 residues) loops (putative EC loops 1, 2, 6, and 7) were not N-glycosylated; however, insertion of EC loop 4 into EC loops 1, 2, or 7, but not 6, resulted in efficient N-glycosylation. Acceptor sites in putative intracellular (IC) loop 5 exhibited a similar pattern of N-glycosylation as EC loop 4, indicating a lumenal disposition during biosynthesis. To be efficiently N-glycosylated, EC loops in polytopic membrane proteins must be larger than 25 residues in size, with acceptor sites located greater than 12 residues away from the preceding TM segment and greater than 14 residues away from the following TM segment. Application of this requirement allowed a significant refinement of the topology of Band 3 including a more accurate mapping of the ends of TM segments. The strict distance dependence for N-glycosylation of loops suggests that TM segments in polytopic membrane proteins are held quite precisely within the translocation machinery during the N-glycosylation process. PMID- 9218474 TI - Characterization of translation initiation factor 5 (eIF5) from Saccharomyces cerevisiae. Functional homology with mammalian eIF5 and the effect of depletion of eIF5 on protein synthesis in vivo and in vitro. AB - Eukaryotic translation initiation factor 5 (eIF5) interacts in vitro with the 40 S initiation complex (40 S.AUG.Met-tRNAf.eIF2.GTP) to mediate the hydrolysis of ribosome-bound GTP. In Saccharomyces cerevisiae, eIF5 is encoded by a single copy essential gene, TIF5, that encodes a protein of 45,346 daltons. To understand the function of eIF5 in vivo, we constructed a conditional mutant yeast strain in which a functional but a rapidly degradable form of eIF5 fusion protein was synthesized from the repressible GAL promoter. Depletion of eIF5 from this mutant yeast strain resulted in inhibition of both cell growth and the rate of in vivo protein synthesis. Analysis of the polysome profiles of eIF5-depleted cells showed greatly diminished polysomes with simultaneous increase in free ribosomes. Furthermore, lysates of cells depleted of eIF5 were dependent on exogenously added yeast eIF5 for efficient translation of mRNAs in vitro. This is the first demonstration that the TIF5 gene encodes a protein involved in initiation of translation in eukaryotic cells. Additionally, we show that rat eIF5 can functionally substitute yeast eIF5 in translation of mRNAs in vitro as well as in complementing in vivo a genetic disruption in the chromosomal copy of TIF5. PMID- 9218475 TI - A cryptic DNA binding domain at the COOH terminus of TFIIIB70 affects formation, stability, and function of preinitiation complexes. AB - TFIIIC-dependent assembly of yeast TFIIIB on class III genes unmasks a high avidity of TFIIIB for DNA. TFIIIB contains TATA-binding protein (TBP), TFIIIB90/B", and TFIIIB70/Brf1, which is homologous to TFIIB. Using limited proteolysis, we have found that the COOH terminus of TFIIIB70 (residues 510-596) forms a protease-resistant domain that binds DNA tightly as seen by Southwestern, DNase I footprinting, and gel shift assays. Consistent with a role for this DNA binding activity, preinitiation complexes were formed less efficiently with truncated TFIIIB70 lacking the COOH-terminal domain and displayed an increased sensitivity to heparin. B' (TFIIIB70 + TBP).TFIIIC.DNA complexes were also particularly unstable. In addition, TFIIIB.TFIIIC.DNA complexes containing truncated TFIIIB70 were impaired in promoting transcription initiation. PMID- 9218476 TI - Phosphorylation controls the three-dimensional structure of plant light harvesting complex II. AB - The most abundant chlorophyll-binding complex in plants is the intrinsic membrane protein light-harvesting complex II (LHC II). LHC II acts as a light-harvesting antenna and has an important role in the distribution of absorbed energy between the two photosystems of photosynthesis. We used spectroscopic techniques to study a synthetic peptide with identical sequence to the LHC IIb N terminus found in pea, with and without the phosphorylated Thr at the 5th amino acid residue, and to study both forms of the native full-length protein. Our results show that the N terminus of LHC II changes structure upon phosphorylation and that the structural change resembles that of rabbit glycogen phosphorylase, one of the few phosphoproteins where both phosphorylated and non-phosphorylated structures have been solved. Our results indicate that phosphorylation of membrane proteins may regulate their function through structural protein-protein interactions in surface-exposed domains. PMID- 9218477 TI - Isolation and molecular cloning of wortmannin-sensitive bovine type III phosphatidylinositol 4-kinases. AB - Agonist-sensitive phosphoinositide pools are maintained by recently-identified wortmannin (WT)-sensitive phosphatidylinositol (PI) 4-kinase(s) (Nakanishi, S., Catt, K. J., and Balla, T. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 5317-5321). Two loosely membrane-associated WT-sensitive type III PI 4-kinases were isolated from bovine adrenal cortex as [3H]WT-labeled 110- and 210-kDa proteins. Based on peptide sequences from the smaller enzyme, a 3. 9-kilobase pair (kb) cDNA with an open reading frame encoding a 90-kDa protein was isolated from a bovine brain cDNA library. Expression of this cDNA in COS-7 cells yielded a 110-kDa protein with WT-sensitive PI 4-kinase activity. Northern blot analysis of a human mRNA panel showed a single approximately 3.8-kb transcript. Peptide sequences obtained from the 210-kDa enzyme corresponded to those of a recently described rat 230-kDa PI 4-kinase. A 6.5-kb cDNA containing an open reading frame of 6129 nucleotides that encoded a 230-kDa protein, was isolated from brain cDNA. Northern blot analysis of human mRNA revealed a major 7.5-kb transcript. The molecular cloning of these novel WT-sensitive type III PI 4-kinases will allow detailed analysis of their signaling and other regulatory functions in mammalian cells. PMID- 9218478 TI - E2F-mediated growth regulation requires transcription factor cooperation. AB - Previous studies have indicated that the presence of an E2F site is not sufficient for G1/S phase transcriptional regulation. For example, the E2F sites in the E2F1 promoter are necessary, but not sufficient, to mediate differential promoter activity in G0 and S phase. We have now utilized the E2F1 minimal promoter to test several hypotheses that could account for these observations. To test the hypothesis that G1/S phase regulation is achieved via E2F-mediated repression of a strong promoter, a variety of transactivation domains were brought to the E2F1 minimal promoter. Although many of these factors caused increased promoter activity, growth regulation was not observed, suggesting that a general repression model is incorrect. However, constructs having CCAAT or YY1 sites or certain GC boxes cloned upstream of the E2F1 minimal promoter displayed E2F site-dependent regulation. Further analysis of the promoter activity suggested that E2F requires cooperation with another factor to activate transcription in S phase. However, we found that the requirement for E2F to cooperate with additional factors to achieve growth regulation could be relieved by bringing the E2F1 activation domain to the promoter via a Gal4 DNA binding domain. Our results suggest a model that explains why some, but not all, promoters that contain E2F sites display growth regulation. PMID- 9218479 TI - Identification of functional elements in the promoter region of the human gene for thymidylate synthase and nuclear factors that regulate the expression of the gene. AB - To identify the essential motifs of the promoter of the human gene for thymidylate synthase (TS), we constructed a set of deletion mutants from the 5' terminal region of the human TS gene. From the results of assays of the expression of chloramphenicol acetyltransferase (CAT), we identified two functional elements with positive effects on the promoter activity: a CACCC box (CCACACCC) and an Sp1-binding motif (GAGGCGGA) that was homologous to the Sp1 binding site in the mouse TS gene. In addition, negative regulatory sequences were identified between the two positive elements and in the region upstream of the CACCC box. The results of gel mobility shift analyses suggested that Sp1 binds to the Sp1-binding motif of the human TS gene promoter and that multiple nuclear factors that are related to Sp1 bind to the CACCC box. Furthermore, the binding of Sp1 to mutated Sp1-binding motifs in the promoter region of the human TS gene was correlated with the promoter activity, as measured by the CAT assay. Therefore, the Sp1 motif seems to be a major contributor to the basic promoter activity of the human TS gene, although multiple positive and negative regulatory elements are involved in the regulated expression of this gene. PMID- 9218480 TI - Phosphorylation at conserved carboxyl-terminal hydrophobic motif regulates the catalytic and regulatory domains of protein kinase C. AB - Mature protein kinase C is phosphorylated at a conserved carboxyl-terminal motif that contains a Ser (or Thr) bracketed by two hydrophobic residues; in protein kinase C betaII, this residue is Ser-660 (Keranen, L. M., Dutil, E. M., and Newton, A. C. (1995) Curr. Biol. 5, 1394-1403). This contribution examines how negative charge at this position regulates the function of protein kinase C. Specifically, Ser-660 in protein kinase C betaII was mutated to Ala or Glu and the enzyme's stability, membrane interaction, Ca2+ regulation, and kinetic parameters were compared with those of wild-type protein phosphorylated at residue 660. Negative charge at this position had no significant effect on the enzyme's diacylglycerol-stimulated membrane interaction nor the conformational change accompanying membrane binding. In contrast, phosphate caused a 10-fold increase in the enzyme's affinity for Ca2+ and a comparable increase in its affinity for phosphatidylserine, two interactions that are mediated by the C2 domain. Negative charge also increased the protein's thermal stability and decreased its Km for ATP and peptide substrate. These data indicate that phosphorylation at the extreme carboxyl terminus of protein kinase C structures the active site so that it binds ATP and substrate with higher affinity and structures determinants in the regulatory region enabling higher affinity binding of Ca2+. The motif surrounding Ser-660 in protein kinase C betaII is found in a number of other kinases, suggesting interactions promoted by phosphorylation of the carboxyl terminus may provide a general mechanism for stabilizing kinase structure. PMID- 9218481 TI - DNA binding characteristics of CrtJ. A redox-responding repressor of bacteriochlorophyll, carotenoid, and light harvesting-II gene expression in Rhodobacter capsulatus. AB - Previous genetic analysis indicated that the photosynthesis gene cluster from Rhodobacter capsulatus coded for the transcription factor, CrtJ, that is responsible for aerobic repression of bacteriochlorophyll, carotenoid, and light harvesting-II gene expression. In this study, we have heterologously overexpressed and purified CrtJ to homogeneity and shown by gel mobility shift assays that CrtJ is biologically active. DNase I footprint analysis confirms molecular genetic studies by showing that CrtJ binds to conserved palindromic sequences that overlap the -10 and -35 promoter regions of the bchC operon. Graphs of the percentage of DNA bound versus protein concentration show sigmoidal curves, which is highly indicative of cooperative binding of CrtJ to the two palindromic sites. A binding constant for interaction of CrtJ with the palindrome that spans the -10 region was calculated to be 4.8 x 10(-9) M, whereas affinity for the palindrome that spans the -35 region was found to be 2.9 x 10(-9) M. Binding of CrtJ to the bchC promoter region was also found to be redox-sensitive, with CrtJ exhibiting a 4.5-fold higher binding affinity under oxidizing versus reducing conditions. PMID- 9218482 TI - The regulation of the cGMP-binding cGMP phosphodiesterase by proteins that are immunologically related to gamma subunit of the photoreceptor cGMP phosphodiesterase. AB - The cGMP phosphodiesterase from retinal rods (PDE-6) is an alphabetagamma2 heterotetramer. The alpha and beta subunits contain catalytic sites for cGMP hydrolysis, whereas the gamma subunits serve as a protein inhibitor of the enzyme. Visual excitation of photoreceptors enables the activated GTP-bound form of the G-protein transducin to remove the inhibitory action of the gamma subunit, thereby triggering PDE-6 activation. The type 5 phosphodiesterase (PDE-5) isoform shares a number of similar characteristics with PDE-6, including binding of cGMP to noncatalytic sites, the cyclic nucleotide specificity, and inhibitor sensitivities. Although the functional role of PDE-5 remains unclear, it has been shown to be activated by protein kinase A (PKA) (Burns, F., Rodger, I. W. & Pyne, N. J. (1992) Biochem. J. 283, 487-491). Here we report that both the recombinant gamma subunit and a peptide corresponding to amino acids 24-46 in this protein inhibited the activation of PDE-5 by PKA. Furthermore, immunoblotting airway smooth muscle membranes with a specific antibody against amino acids 24-46 of the PDE-6 gamma subunit identified two major immunoreactive small molecular mass proteins of 14 and 18 kDa (p14 and p18). These appear to form a complex with PDE 5, because PDE activity was immunoprecipitated using antibody against the PDE-6 gamma subunit. p14 and p18 were also substrates for phosphorylation by a unidentified kinase that was stimulated by a pertussis toxin-sensitive G-protein. Phosphorylation of p14/p18 in membranes treated with guanine nucleotides correlated with a concurrent reduction in the activation of PDE-5 by PKA. We suggest that p14 and p18 share an epitope common to PDE-6 gamma and that this region may interact with PDE-5 to prevent its activation by PKA. PMID- 9218483 TI - Critical role of glutamate in a central leucine-rich repeat of decorin for interaction with type I collagen. AB - The chondroitin/dermatan sulfate proteoglycan decorin is known to interact via its core protein with fibrillar collagens, thereby influencing the kinetics of fibril formation and the final diameter of the fibrils. To define the binding site(s) for type I collagen along the core protein, which is mainly composed of leucine-rich repeat structures, decorin cDNAs were constructed and expressed in human kidney 293 cells. The constructs encoded (i) C-terminally truncated molecules, (ii) core proteins with deletions of selected leucine-rich repeats, or (iii) various point mutations. The deletion of the sixth leucine-rich repeat Met176-Lys201 and the mutation E180K drastically interfered with the binding to reconstituted type I collagen fibrils. In contrast, the deletion of the seventh repeat Leu202-Ser222 led at the most to a marginally impaired binding, although the secretion of this proteoglycan was abnormally low. Decorin with two other point mutations in the sixth leucine-rich repeat, Lys187 --> Gln and Lys200 --> Gln, respectively, bound type I collagen either normally or even better than the normal recombinant proteoglycan. These data suggest that a major collagen-binding site of decorin is located within the sixth leucine-rich repeat and that glutamate-180 within this repeat is of special importance for ionic interactions between the two matrix components. PMID- 9218484 TI - Thiol compounds interact with nitric oxide in regulating heme oxygenase-1 induction in endothelial cells. Involvement of superoxide and peroxynitrite anions. AB - Thiols are very important antioxidants that protect cells against oxidative insults. Recently, a different and new physiological role has been defined for these compounds because of their involvement in nitric oxide (NO) binding and transport in biological systems. In view of these characteristics, we examined the effect of thiols and NO on the expression of the inducible form of heme oxygenase (HO-1), a stress protein that degrades heme to carbon monoxide and biliverdin. Cultured bovine aortic endothelial cells exposed to the NO donors sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine (SNAP) resulted in increased heme oxygenase activity and HO-1 expression. Co-incubation with N acetylcysteine, a precursor of glutathione synthesis, significantly attenuated heme oxygenase induction by SNP and SNAP, and a reduction in heme oxygenase activity was also observed when cells were preincubated with N-acetylcysteine for 16 h prior to exposure to NO donors. This effect appears to be associated with NO stabilization by thiols through the formation of S-nitrosothiols. Hydroxocobalamin, a specific NO scavenger, significantly decreased endothelial heme oxygenase activity, indicating a direct involvement of NO released by NO donors to regulate the expression of this stress protein. Moreover, superoxide anion (O-2) and its reaction product with NO, peroxynitrite (ONOO-), were found to partially contribute to the observed NO-mediated activation of endothelial heme oxygenase. Thus, we suggest the existence of a dynamic equilibrium among free NO, O-2, and endogenous glutathione, which might constitute an interactive signaling mechanism modulating stress and adaptive responses in tissues. PMID- 9218485 TI - Syndecan-1 expression is down-regulated during myoblast terminal differentiation. Modulation by growth factors and retinoic acid. AB - Syndecan-1 is an integral membrane proteoglycan involved in the interaction of cells with extracellular matrix proteins and growth factors. It is transiently expressed in several condensing mesenchymal tissues after epithelial induction. In this study we evaluated the expression of syndecan-1 during skeletal muscle differentiation. The expression of syndecan-1 as determined by Northern blot analyses and immunofluorescence microscopy is down-regulated during differentiation. The transcriptional activity of a syndecan-1 promoter construct is also down-regulated in differentiating muscle cells. The decrease in syndecan 1 gene expression is not dependent on the presence of E-boxes, binding sites for the MyoD family of transcription factors in the promoter region, or myogenin expression. Deletion of the region containing the E-boxes or treatment of differentiating cells with sodium butyrate, an inhibitor of myogenin expression, had no effect on syndecan-1 expression. Basic fibroblast growth factor and transforming growth factor type beta, which are inhibitors of myogenesis, had little effect on syndecan-1 expression. When added together, however, they induced syndecan-1 expression. Retinoic acid, an inducer of myogenesis, inhibited syndecan-1 expression and abolished the effect of the growth factors. These results indicate that syndecan-1 expression is down-regulated during myogenesis and that growth factors and retinoic acid modulate syndecan-1 expression by a mechanism that is independent of myogenin. PMID- 9218486 TI - The acidic carboxyl terminus of the bacteriophage T7 gene 4 helicase/primase interacts with T7 DNA polymerase. AB - The gene 4 proteins of bacteriophage T7 provide both primase and helicase activities at the replication fork. Efficient DNA replication requires that the functions of the gene 4 protein be coordinated with the movement of the T7 DNA polymerase. We show that a carboxyl-terminal domain of the gene 4 protein is required for interaction with T7 DNA polymerase during leading strand DNA synthesis. The carboxyl terminus of the gene 4 protein is highly acidic: of the 17 carboxyl-terminal amino acids 7 are negatively charged. Deletion of the coding region for these 17 residues results in a gene 4 protein that cannot support the growth of T7 phage. The purified mutant gene 4 protein has wild-type levels of both helicase and primase activities; however, DNA synthesis catalyzed by T7 DNA polymerase on a duplex DNA substrate is stimulated by this mutant protein to only about 5% of the level of synthesis obtained with wild-type protein. The mutant gene 4 protein can form hexamers and bind single-stranded DNA, but as determined by native PAGE analysis, the protein cannot form a stable complex with the DNA polymerase. The mutant gene 4 protein can prime DNA synthesis normally, indicating that for lagging strand synthesis a different set of helicase/primase DNA polymerase interactions are involved. These findings have implications for the mechanisms coupling leading and lagging strand DNA synthesis at the T7 replication fork. PMID- 9218487 TI - Hin-mediated inversion on positively supercoiled DNA. AB - Hin recombinase requires negatively supercoiled DNA for an efficient inversion. We have generated positively supercoiled plasmid DNA using reverse gyrase from Sulfolobus shibatae and subjected it to the Hin-mediated inversion reaction. Both Hin and Fis showed the same DNA binding activity regardless of the superhelical handedness of the substrate plasmid. However, inversion activity on positively supercoiled DNA was less than 1% of negatively supercoiled DNA. Assays designed to probe steps in inversion, showed that on positively supercoiled DNA, Hin was able to cleave the recombination sites with the same efficiency shown on negatively supercoiled DNA but was not able to exchange the cleaved DNA. Based on the theoretical differences between positive and negative supercoiling, our data may suggest that unwinding of the double helix at recombination sites is needed after DNA cleavage for strand exchange to occur. PMID- 9218488 TI - Interaction of the nuclear matrix-associated region (MAR)-binding proteins, SATB1 and CDP/Cux, with a MAR element (L2a) in an upstream regulatory region of the mouse CD8a gene. AB - Matrix-associated regions (MARs), AT-rich DNA segments that have an affinity for the nuclear matrix, have been shown to play a role in transcriptional regulation of eukaryotic genes. The present study demonstrates that a DNA element, called L2a, which has been implicated in the transcriptional regulation of the mouse CD8a gene encoding an important T cell coreceptor, is a MAR. Moreover, the identities of two nuclear proteins, L2a-P1 and L2a-P2, previously shown to bind to the L2a element, have been determined. The L2a-P1 protein found to be present in all CD8-positive T cell lines tested is SATB1, a known MAR-binding protein. The widely expressed L2a-P2 protein is CDP/Cux, a MAR-binding protein that has been associated with repression of gene transcription. Interaction of both proteins with the L2a element was studied using the missing nucleoside approach, DNase I footprinting, and electrophoretic mobility shift assays with wild type and mutant L2a elements. The data suggest that CDP/Cux bound to the L2a element is displaced by binding of SATB1 and the accompanying conformational change in the DNA lying between the primary binding sites of SATB1 and CDP/Cux. We suggest that displacement of CDP/Cux by SATB1 favors transcription of the CD8a gene, possibly by enhancing or altering its association with the nuclear matrix. PMID- 9218489 TI - Selective induction of heparin-binding epidermal growth factor-like growth factor by methylglyoxal and 3-deoxyglucosone in rat aortic smooth muscle cells. The involvement of reactive oxygen species formation and a possible implication for atherogenesis in diabetes. AB - Methylglyoxal (MG) and 3-deoxyglucosone (3-DG), reactive dicarbonyl metabolites in the glyoxalase system and glycation reaction, respectively, selectively induced heparin-binding epidermal growth factor (HB-EGF)-like growth factor mRNA in a dose- and time-dependent manner in rat aortic smooth muscle cells (RASMC). A nuclear run-on assay revealed that the dicarbonyl may regulate expression of HB EGF at the transcription level. The dicarbonyl also increased the secretion of HB EGF from RASMC. However, platelet-derived growth factor, another known growth factor of smooth muscle cells (SMC), was not induced by both dicarbonyls. The dicarbonyl augmented intracellular peroxides prior to the induction of HB-EGF mRNA as judged by flow cytometric analysis using 2',7'-dichlorofluorescin diacetate. N-Acetyl-L-cysteine and aminoguanidine suppressed both dicarbonyl increased HB-EGF mRNA and intracellular peroxide levels in RASMC. DL-Buthionine (S, R)-sulfoximine increased the levels of 3-DG-induced HB-EGF mRNA. Furthermore, hydrogen peroxide alone also induced HB-EGF mRNA in RASMC. These results indicate that MG and 3-DG induce HB-EGF by increasing the intracellular peroxide levels. In addition, the pretreatment with 12-O-tetra-decanoylphorbol-13-acetate failed to alter dicarbonyl-induced HB-EGF mRNA expression in RASMC, suggesting that the signal transducing mechanism is not mediated by protein kinase C. Since HB-EGF is known as a potent mitogen for smooth muscle cells and is abundant in atherosclerotic plaques, the induction of HB-EGF by MG and 3-DG, as well as the concomitant increment of intracellular peroxides, may trigger atherogenesis during diabetes. PMID- 9218490 TI - Biosynthesis of mycobacterial lipoarabinomannan. AB - The mycobacterial lipoglycans, lipomannan (LM) and lipoarabinomannan (LAM), are potent immunomodulators in tuberculosis and leprosy. Little is known of their biosynthesis, other than being based on phosphatidylinositol (PI), and they probably originate in the phosphatidylinositol mannosides (PIMs; PIMans). A novel form of cell-free incubation involving in vitro and in situ labeling with GDP [14C]Man of the polyprenyl-P-mannoses (C35/C50-P-Man) and the simpler PIMs of mycobacterial membranes, reisolation of the [14C]Man-labeled membranes, and in situ chase demonstrated the synthesis of a novel alpha(1-->6)-linked linear form of LM at the expense of the C35/C50-P-Man. There was little or no synthesis under these conditions of PIMan5 with its terminal alpha(1-->2)Man unit or the mature LM or LAM with copious alpha(1-->2)Man branching. Synthesis of the linear LM, but not of the simpler PIMan2, was susceptible to amphomycin, a lipopeptide antibiotic that specifically inhibits polyprenyl-P-requiring translocases. A mixture of P[3H]I and P[3H]IMan2 was incorporated into the linear LM, supporting other evidence that, like the PIMs, LM and LAM, it is a lipid-linked mannooligosaccharide and a new member of the mycobacterial glycosylphosphatidylinositol lipoglycan/glycolipid class. Hence, the simpler PIMs originate in PI and GDP-Man, but further growth of the linear backbone emanates from C35-/C50-P-Man and is amphomycin-sensitive. The origin of the alpha(1- >2)Man branches of mature PIMan5, LM, and LAM is not known at this time but is probably GDP-Man. PMID- 9218491 TI - Analysis of the functional domain of the rat liver mitochondrial import receptor Tom20. AB - Tom20 is an outer mitochondrial membrane protein and functions as a component of the import receptor complex for the cytoplasmically synthesized mitochondrial precursor proteins. It consists of the N-terminal membrane-anchor segment, the tetratricopeptide repeat (TPR) motif, a charged amino acids-rich linker segment between the membrane anchor and the TPR motif, and the C-terminal acidic amino acid cluster. To assess the functional significance of these segments in mammalian Tom20, we cloned rat Tom20 and expressed mutant rat Tom20 proteins in Deltatom20 yeast cells and examined their ability to complement the defects of respiration-driven growth and mitochondrial protein import. Tom20N69, a mutant consisting of the membrane anchor and the linker segments, was targeted to mitochondria and complemented the growth and import defects as efficiently as wild-type Tom20, whereas a mutant lacking the linker segment did not. In vitro protein import into mitochondria isolated from the complemented yeast cells revealed that the precursor targeted to yeast Tom70 was efficiently imported into the mitochondria via rat Tom20N69. Thus the linker segment is essential for the function of rat Tom20, whereas the TPR motif and the C-terminal acidic amino acids are not. PMID- 9218492 TI - Regulation of the epithelial brush border Na+/H+ exchanger isoform 3 stably expressed in fibroblasts by fibroblast growth factor and phorbol esters is not through changes in phosphorylation of the exchanger. AB - The epithelial brush border Na+/H+ exchanger isoform 3 (NHE3) is regulated by growth factors and protein kinases. When stably expressed in PS120 fibroblasts, NHE3 is stimulated by serum and fibroblast growth factor (FGF) and inhibited by phorbol esters. To examine the role of phosphorylation of NHE3 in growth factor/protein kinase regulation, NHE3 was C-terminally tagged with an 11-amino acid epitope of the vesicular stomatitis virus glycoprotein (VSVG) and stably expressed in Na+/H+ exchanger null PS120 fibroblasts (PS120/NHE3V). NHE3V was regulated by serum, FGF, and phorbol ester in a manner identical to wild type non VSVG-tagged NHE3. Phosphorylation of NHE3V was evaluated via immunoprecipitation with anti-VSVG antibody after in vivo labeling of PS120/NHE3V cells with [32P]orthophosphate. NHE3V was phosphorylated under basal conditions. However, FGF and PMA, under conditions in which these agonists regulate NHE3V, altered neither the amount of phosphorylation of NHE3V as analyzed by one-dimensional SDS polyacrylamide gel electrophoresis and autoradiography nor two-dimensional phosphopeptide maps of tryptic digests of NHE3V. In contrast, while changes in NHE3V phosphorylation were not observed with serum exposure by one-dimensional SDS-polyacrylamide gel electrophoresis, two-dimensional studies showed increases in two phosphopeptides. Under all these conditions, phosphoamino acid analysis showed that NHE3V was phosphorylated only on serine residues. By cell surface protein biotinylation studies under basal conditions, at least 27% of the NHE3V was expressed on the cell surface. To further analyze the phosphorylation status of the surface and intracellular forms of NHE3V under basal conditions and determine whether the amount of phosphorylation of the surface form changes upon serum, FGF, and PMA regulation, the surface form of NHE3V was separated from intracellular form by biotinylation/avidin-agarose precipitation. Under basal conditions, both intracellular and surface forms of NHE3V were phosphorylated. However, the amount of phosphorylation of the surface form of NHE3V did not change upon stimulation by serum and FGF and inhibition by PMA based on one dimensional SDS-polyacrylamide gel electrophoresis and autoradiography. Thus, we conclude that when expressed in PS120 cells, while NHE3 is a phosphoprotein under basal conditions, its regulation by FGF and PMA is not by changes in the phosphorylation of NHE3, while regulation by serum may involve changes in its phosphorylation. Regulation of NHE3 probably involves intermediate associated regulatory proteins. The function of basal phosphorylation of NHE3 is not known. PMID- 9218493 TI - Molecular cloning and expression of cDNA encoding 3alpha,7alpha,12alpha trihydroxy-5beta-chole stanoyl-CoA oxidase from rabbit liver. AB - The steroid side chain cleavage in bile acid formation is catalyzed by liver peroxisomal enzymes (Pedersen, J. I. and Gustafsson, J. (1980) FEBS Lett. 121, 345-348; Kase, F., Bjorkhem, I., and Pedersen, J. I. (1983) J. Lipid Res. 24, 1560-1567). We here describe the cloning and sequencing of a cDNA coding the first of these enzymes, a 3alpha,7alpha,12alpha-trihydroxy-5beta-choles tanoyl CoA oxidase (THCA-CoA oxidase) from rabbit liver peroxisomes. After tryptic digestion of purified protein in a polyacrylamide gel, five peptides were isolated and sequenced. Using two oligonucleotides deduced from the amino acid sequence data, two overlappping clones were isolated from a rabbit liver cDNA library, which together made up a unique cDNA sequence of 2139 base pairs. It contained an open reading frame of 2046 base pairs encoding a protein of 681 amino acids with a molecular mass of 76,209 daltons. All five peptides could be localized within the sequence. Transfection of COS cells with the coding part of the cDNA resulted in a significant expression of THCA-CoA oxidase activity. We were not able to demonstrate 3alpha, 7alpha-dihydroxy-5beta-cholestanoyl-CoA oxidase activity under the same conditions. The obtained sequence showed 73.6% similarity with a proposed rat THCA-CoA oxidase and 81% similarity with a recently reported human branched chain acyl-CoA oxidase, indicating that these three proteins represent the same enzyme. The similarity with rat palmitoyl-CoA oxidase was 41.8%. The C-terminal tripeptide of the protein was SNL, a previously undescribed variant of the main class of peroxisomal targeting signals. Northern blot analysis revealed that the gene is transcribed in liver and kidney, and the major mRNA fraction had a size of approximately 2.6 kilobase pairs. PMID- 9218494 TI - Two distinct factor-binding DNA elements in cardiac myosin light chain 2 gene are essential for repression of its expression in skeletal muscle. Isolation of a cDNA clone for repressor protein Nished. AB - The expression of the cardiac myosin light chain 2 (MLC2) gene is repressed in skeletal muscle as a result of the negative regulation of its transcription. Two regulatory elements, the cardiac specific sequence (CSS) located upstream (-360 base pairs) and a downstream negative modulatory sequence (NMS), which function in concert with each other, are required for repression of the MLC2 promoter activity in skeletal muscle. Individually, CSS and NMS have no effect. Transient transfection analysis with recombinant plasmids indicated that CSS- and NMS mediated repression of transcription is position- and orientation-dependent and is transferable to heterologous promoters. A minimal conserved motif, GAAG/CTTC, present in both CSS and NMS, is responsible for repression as the mutation in the core CTTC sequence alone was sufficient to abrogate its repressor activity. The DNA binding assay by gel mobility shift analysis revealed that one of the two complexes, CSSBP2, is significantly enriched in embryonic skeletal muscle relative to cardiac muscle. In extracts from adult skeletal muscle, where the cardiac MLC2 expression is suppressed, both complexes, CSSBP1 and CSSBP2, were present, whereas the cardiac muscle extracts contained CSSBP1 alone, suggesting that the protein(s) in the CSSBP2 complex accounts for the negative regulation of cardiac MLC2 in skeletal muscle. A partial cDNA clone (Nished) specific for the candidate repressor factor was isolated by expression screening of the skeletal muscle cDNA library by multimerized CSS-DNA as probe. The recombinant Nished protein binds to the CSS-DNA, but not to DeltaCSS-DNA where the core CTTC sequence was mutated. The amino acid sequence of Nished showed a significant structural similarity to the sequence of transcription factor "runt," a known repressor of gap and pair-rule gene expression in Drosophila. PMID- 9218495 TI - Oligomerization of the Fes tyrosine kinase. Evidence for a coiled-coil domain in the unique N-terminal region. AB - The c-fes proto-oncogene encodes a non-receptor tyrosine kinase (Fes) that has been implicated in cytokine receptor signal transduction and myeloid differentiation. Previous work from our laboratory has shown that Fes autophosphorylates via an intermolecular mechanism more commonly associated with growth factor receptor tyrosine kinases. Analysis of the Fes amino acid sequence with the COILS algorithm indicates that the N-terminal region of the protein has a very high probability of forming coiled-coil structures often associated with oligomeric proteins. These findings suggest that oligomerization may be a prerequisite for trans-autophosphorylation and activation of Fes. To establish whether the active form of Fes is oligomeric, we performed gel-filtration experiments with recombinant Fes and found that it eluted as a single symmetrical peak of approximately 500 kDa. No evidence of the monomeric, 93-kDa form of the protein was observed. Deletion of the unique N-terminal domain (amino acids 1 450, including the coiled-coil homology region) completely abolished the formation of oligomers. Furthermore, co-precipitation assays demonstrated that an immobilized glutathione S-transferase fusion protein containing the Fes N terminal region bound to full-length Fes but not to a mutant lacking the N terminal region. Similarly, a recombinant Fes N-terminal domain protein was readily cross-linked in vitro, whereas the SH2 and kinase domains were refractory to cross-linking. Incubation of wild-type Fes with a kinase-inactive Fes mutant or with the isolated N-terminal region suppressed Fes autophosphorylation in vitro, suggesting that oligomerization may be essential for autophosphorylation of full-length Fes. The presence of an oligomerization function in the Fes family of tyrosine kinases suggests a novel mechanism for non-receptor protein-tyrosine kinase regulation. PMID- 9218497 TI - Extracellular interaction of the voltage-dependent Ca2+ channel alpha2delta and alpha1 subunits. AB - The role of the extracellular domain of the voltage-dependent Ca2+ channel alpha2delta subunit in assembly with the alpha1C subunit was investigated. Transiently transfected tsA201 cells processed the alpha2delta subunit properly as disulfide linkages and cleavage sites between the alpha2 and delta subunits were shown to be similar to native channel protein. Coimmunoprecipitation experiments demonstrated that in the absence of delta subunits, alpha2 subunits do not assemble with alpha1 subunits. Furthermore, the transmembrane and cytoplasmic sequences in delta can be exchanged with those of an unrelated protein without any effect on the association between the alpha2delta and alpha1 proteins. Extracellular domains of the alpha2delta subunit are also shown to be responsible for increasing the binding affinity of [3H]PN200-110 (isopropyl-4 (2,1, 3-benzoxadiazol-4-yl)-1,4-dihydro-2, 6-dimethyl-5-([3H]methoxycarbonyl) pyridine-3-carboxylate) for the alpha1C subunit. Investigation of the corresponding interaction site on the alpha1 subunit revealed that although tryptic peptides containing repeat III of native alpha1S subunit remain in association with the alpha2delta subunit during wheat germ agglutinin chromatography, repeat III by itself is not sufficient for assembly with the alpha2delta subunit. Our results suggest that the alpha2delta subunit likely interacts with more than one extracellular loop of the alpha1 subunit. PMID- 9218496 TI - Two potent nuclear localization signals in the gut-enriched Kruppel-like factor define a subfamily of closely related Kruppel proteins. AB - The gut-enriched Kruppel-like factor (GKLF) is a newly identified transcription factor that contains three C2H2 Kruppel-type zinc fingers. Previous immunocytochemical studies indicate that GKLF is exclusively localized to the nucleus. To identify the nuclear localization signal (NLS) within GKLF, cDNA constructs with various deletions in the coding region of GKLF were generated and analyzed by indirect immunofluorescence in transfected COS-1 cells. In addition, constructs fusing regions representing putative NLSs of GKLF to green fluorescent protein (GFP) were generated and examined by fluorescence microscopy in similarly transfected cells. The results indicate that GKLF contains two potent, independent NLSs: one within the zinc fingers and the other in a cluster of basic amino acids (called 5' basic region) immediately preceding the first zinc finger. In comparison, putative NLSs within the zinc fingers and the 5' basic region of a related Kruppel protein, zif268/Egr-1, are relatively less efficient in their ability to translocate GFP into the nucleus. A search in the protein sequence data base revealed that despite the existence of numerous Kruppel proteins, only two, the lung Kruppel-like factor (LKLF) and the erythroid Kruppel-like factor (EKLF), exhibit similar NLSs to those of GKLF. These findings indicate that GKLF, LKLF, and EKLF are members of a subfamily of closely related Kruppel proteins. PMID- 9218498 TI - Perspectives series: host/pathogen interactions. Invasion and intracellular sorting of bacteria: searching for bacterial genes expressed during host/pathogen interactions. PMID- 9218499 TI - Oxidized or acetylated low density lipoproteins are rapidly cleared by the liver in mice with disruption of the scavenger receptor class A type I/II gene. AB - Oxidized low density lipoprotein (LDL) and acetyl LDL are recognized by the scavenger receptor class A type I/II (SR-AI/II) on macrophages and liver endothelial cells. Several investigators have suggested that there are additional receptors specific for oxidized LDL, but characterization of these alternate receptors for oxidized LDL and evaluation of their quantitative importance in uptake of oxidized LDL has been difficult because of overlapping ligand specificity with SR-AI/II. The purpose of this study was to determine the importance of SR-AI/II in the removal of modified LDL from the bloodstream in vivo. The clearance rate of oxidized LDL from plasma in normal mice was very rapid, and > 90% of injected dose was removed from the blood within 5 min. Clearance rates of oxidized LDL were equally high in SR-AI/II knockout mice, indicating that this receptor is not required for removal of oxidized LDL from plasma. Surprisingly, there was no difference in the clearance rate of acetyl LDL in wild-type and SR-AI/II knockout animals. The plasma clearance of radioiodinated acetyl LDL was almost fully blocked by a 50-fold excess of unlabeled acetyl LDL, but the latter only inhibited oxidized LDL clearance by approximately 5%. Both modified LDLs were cleared mostly by the liver, and there was no difference in the tissue distribution of modified LDL in control and knockout mice. Studies in isolated nonparenchymal liver cells showed that Kupffer cells accounted for most of the uptake of oxidized LDL. Extensively oxidized LDL and LDL modified by exposure to fatty acid peroxidation products were efficient competitors for the uptake of labeled oxidized LDL by SR-AI/II-deficient Kupffer cells, while acetyl LDL and malondialdehyde-modified LDL were relatively poor competitors. PMID- 9218500 TI - Compartmentalization of angiotensin II generation in the dog heart. Evidence for independent mechanisms in intravascular and interstitial spaces. AB - Angiotensin-converting enzyme inhibitors have beneficial effects that are presumably mediated by decreased angiotensin II (ANG II) production. In this study, we measure for the first time ANG I and ANG II levels in the interstitial fluid (ISF) space of the heart. ISF and aortic plasma ANG I and II levels were obtained at baseline, during intravenous infusion of ANG I (5 microM, 0.1 ml/min, 60 min), and during ANG I + the angiotensin-converting enzyme inhibitor captopril (cap) (2.5 mM, 0.1 ml/min, 60 min) in six anesthetized open-chested dogs. ISF samples were obtained using microdialysis probes inserted into the left ventricular myocardium (3-4 probes/dog). ANG I increased mean arterial pressure from 102+/-3 (SEM) to 124+/-3 mmHg (P < 0.01); addition of cap decreased MAP to 95+/-3 mmHg (P < 0.01). ANG I infusion increased aortic plasma ANG I and ANG II (pg/ml) (ANG I = 101+/-129 to 370+/-158 pg/ml, P < 0.01; and ANG II = 22+/-40 to 466+/-49, P < 0.01); addition of cap further increased ANG I (1,790+/-158, P < 0.01) and decreased ANG II (33+/-49, P < 0.01). ISF ANG I and ANG II levels (pg/ml) were > 100-fold higher than plasma levels, and did not change from baseline (8,122+/-528 and 6,333+/-677), during ANG I (8,269+/-502 and 6, 139+/ 695) or ANG I + cap (8,753+/-502 and 5,884+/-695). The finding of very high ANG I and ANG II levels in the ISF vs. intravascular space that are not affected by IV ANG I or cap suggests that ANG II production and/or degradation in the heart is compartmentalized and mediated by different enzymatic mechanisms in the interstitial and intravascular spaces. PMID- 9218501 TI - Vitamin D receptor genotype is associated with radiographic osteoarthritis at the knee. AB - Osteoporosis and osteoarthritis are age-related disorders of the skeleton with genetic components. Low bone density is a risk factor for osteoporotic fracture while osteoarthritis is associated with increased bone density. The 1,25 dihydroxyvitamin D3 receptor (VDR) gene locus was previously found to be associated with bone density. We therefore studied the relationship between radiographic osteoarthritis at the knee and VDR genotype in a population-based sample (n = 846), using molecular haplotyping of anonymous intragenic DNA polymorphisms. Radiographic osteoarthritis was defined using the Kellgren score, which is based on the assessment of osteophytes and joint space narrowing (JSN). We show that one VDR haplotype allele is significantly overrepresented in individuals with knee osteoarthritis and associated with a 2.27-fold increased relative risk (95% confidence interval 1.46, 3.52). Adjustment for bone density at the femoral neck did not change these results, indicating that the association is not mediated by bone density. The association appeared to be largely explained by the presence of osteophytes rather than JSN. Our results indicate a role of the VDR gene in the pathogenesis of osteophytes while linkage disequilibrium with another nearby gene, i.e., the collagen type IIa1 gene encoding the most abundant protein in cartilage, might contribute to the association. PMID- 9218502 TI - The subtype 2 (AT2) angiotensin receptor mediates renal production of nitric oxide in conscious rats. AB - The angiotensin AT2 receptor modulates renal production of cyclic guanosine 3',5' monophosphate (cGMP; J. Clin. Invest. 1996. 97:1978-1982). In the present study, we hypothesized that angiotensin II (Ang II) acts at the AT2 receptor to stimulate renal production of nitric oxide leading to the previously observed increase in cGMP. Using a microdialysis technique, we monitored changes in renal interstitial fluid (RIF) cGMP in response to intravenous infusion of the AT2 receptor antagonist PD 123319 (PD), the AT1 receptor antagonist Losartan, the nitric oxide synthase (NOS) inhibitor nitro--arginine-methyl-ester (-NAME), the specific neural NOS inhibitor 7-nitroindazole (7-NI), or Ang II individually or combined in conscious rats during low or normal sodium balance. Sodium depletion significantly increased RIF cGMP. During sodium depletion, both PD and -NAME caused a similar decrease in RIF cGMP. Combined administration of PD and -NAME decreased RIF cGMP to levels observed with PD or -NAME alone or during normal sodium intake. During normal sodium intake, Ang II caused a twofold increase in RIF cGMP. Neither PD nor -NAME, individually or combined, changed RIF cGMP. Combined administration of Ang II and either PD or -NAME produced a significant decrease in RIF cGMP compared with that induced by Ang II alone. Combined administration of Ang II, PD, and -NAME blocked the increase in RIF cGMP produced by Ang II alone. During sodium depletion, 7-NI decreased RIF cGMP, but the reduction of cGMP in response to PD alone or PD combined with 7-NI was greater than with 7-NI alone. During normal sodium intake, 7-NI blocked the Ang II induced increase in RIF cGMP. PD alone or combined with 7-NI produced a greater inhibition of cGMP than did 7-NI alone. During sodium depletion, 7-NI (partially) and -NAME (completely) inhibited RIF cGMP responses to -arginine. These data demonstrate that activation of the renin- angiotensin system during sodium depletion increases renal nitric oxide production through stimulation by Ang II at the angiotensin AT2 receptor. This response is partially mediated by neural NOS, but other NOS isoforms also contribute to nitric oxide production by this pathway. PMID- 9218503 TI - Receptor-mediated regional sympathetic nerve activation by leptin. AB - Leptin is a peptide hormone produced by adipose tissue which acts centrally to decrease appetite and increase energy expenditure. Although leptin increases norepinephrine turnover in thermogenic tissues, the effects of leptin on directly measured sympathetic nerve activity to thermogenic and other tissues are not known. We examined the effects of intravenous leptin and vehicle on sympathetic nerve activity to brown adipose tissue, kidney, hindlimb, and adrenal gland in anesthetized Sprague-Dawley rats. Intravenous infusion of mouse leptin over 3 h (total dose 10-1,000 microg/kg) increased plasma concentrations of immunoreactive murine leptin up to 50-fold. Leptin slowly increased sympathetic nerve activity to brown adipose tissue (+286+/-64% at 1,000 microg/kg; P = 0.002). Surprisingly, leptin infusion also produced gradual increases in renal sympathetic nerve activity (+228+/-63% at 1,000 microg/kg; P = 0.0008). The effect of leptin on sympathetic nerve activity was dose dependent, with a threshold dose of 100 microg/kg. Leptin also increased sympathetic nerve activity to the hindlimb (+287+/-60%) and adrenal gland (388+/-171%). Despite the increase in overall sympathetic nerve activity, leptin did not increase arterial pressure or heart rate. Leptin did not change plasma glucose and insulin concentrations. Infusion of vehicle did not alter sympathetic nerve activity. Obese Zucker rats, known to possess a mutation in the gene for the leptin receptor, were resistant to the sympathoexcitatory effects of leptin, despite higher achieved plasma leptin concentrations. These data demonstrate that leptin increases thermogenic sympathetic nerve activity and reveal an unexpected stimulatory effect of leptin on overall sympathetic nerve traffic. PMID- 9218504 TI - CD4(+) T-lymphocytes mediate ischemia/reperfusion-induced inflammatory responses in mouse liver. AB - The success of orthotopic liver transplantation is dependent on multiple factors including MHC tissue compatibility and ischemic/reperfusion injury. Ischemic/reperfusion (I/R) injury in the liver occurs in a biphasic pattern consisting of both acute phase (oxygen free radical mediated) and subacute phase (neutrophil-mediated) damage. Although numerous studies have given insights into the process of neutrophil recruitment after I/R injury to the liver, the exact mechanism that initiates this subacute response remains undefined. Using a T cell deficient mouse model, we present data that suggests that T-lymphocytes are key mediators of subacute neutrophil inflammatory responses in the liver after ischemia and reperfusion. To this end, using a partial lobar liver ischemia model, we compared the extent of reperfusion injury between immune competent BALB/c and athymic nu/nu mice. Studies evaluating the extent of liver damage as measured by serum transaminases (GPT) demonstrate similar acute (3-6 h) post-I/R responses in these two mouse models. In contrast, the subacute phase (16-20 h) of liver injury, as measured by both serum GPT levels and percent hepatocellular necrosis, was dramatically reduced in T cell-deficient mice as compared with those with an intact immune system. This reduction in liver injury seen in nu/nu mice was associated with a 10-fold reduction in hepatic neutrophil infiltration. Adoptive transfer of T cell-enriched splenocytes from immune competent mice was capable of reconstituting the neutrophil-mediated subacute inflammatory response within T cell-deficient nu/nu mice. Furthermore, in vivo antibody depletion of CD4(+) T-lymphocytes in immune competent mice resulted in a reduction of subacute phase injury and inflammation as measured by serum GPT levels and neutrophil infiltration. In contrast, depletion of CD8(+) T-lymphocytes had no effect on these indexes of subacute inflammation. Kinetic analysis of T cell infiltration in the livers of BALB/c mice demonstrated a fivefold increase in the number of hepatic CD4(+) T-lymphocytes within the first hour of reperfusion with no significant change in the number of CD8(+) T-lymphocytes. In summary, these results implicate CD4(+) T-lymphocytes as key regulators in initiating I/R induced inflammatory responses in the liver. Such findings have implications for therapy directed at the early events in this inflammatory cascade that may prove useful in liver transplantation. PMID- 9218505 TI - Role of nitric oxide in obesity-induced beta cell disease. AB - Here we report that free fatty acid-induced suppression of insulin output in prediabetic Zucker diabetic fatty (ZDF) rats is mediated by NO. When normal islets were cultured in 2 mM FFA, NO production and basal insulin secretion increased slightly. In cultured prediabetic ZDF islets, FFA induced a fourfold greater rise in NO, upregulated mRNA of inducible nitric oxide synthase (iNOS), and reduced insulin output; both nicotinamide and aminoguanidine, which lower NO, prevented the FFA-mediated increase in iNOS mRNA, reduced NO, and minimized the loss of insulin secretion. In vivo nicotinamide or aminoguanidine treatment of prediabetic ZDF rats prevented the iNOS expression in islets and decreased beta cell dysfunction while blocking beta cell destruction and hyperglycemia. We conclude that NO-lowering agents prevent adipogenic diabetes in obese rats. PMID- 9218506 TI - Role of intestinal epithelial cells in the host secretory response to infection by invasive bacteria. Bacterial entry induces epithelial prostaglandin h synthase 2 expression and prostaglandin E2 and F2alpha production. AB - Increased intestinal fluid secretion is a protective host response after enteric infection with invasive bacteria that is initiated within hours after infection, and is mediated by prostaglandin H synthase (PGHS) products in animal models of infection. Intestinal epithelial cells are the first host cells to become infected with invasive bacteria, which enter and pass through these cells to initiate mucosal, and ultimately systemic, infection. The present studies characterized the role of intestinal epithelial cells in the host secretory response after infection with invasive bacteria. Infection of cultured human intestinal epithelial cell lines with invasive bacteria, but not noninvasive bacteria, is shown to induce the expression of one of the rate-limiting enzymes for prostaglandin formation, PGHS-2, and the production of PGE2 and PGF2alpha. Furthermore, increased PGHS-2 expression was observed in intestinal epithelial cells in vivo after infection with invasive bacteria, using a human intestinal xenograft model in SCID mice. In support of the physiologic importance of epithelial PGHS-2 expression, supernatants from bacteria-infected intestinal epithelial cells were shown to increase chloride secretion in an in vitro model using polarized epithelial cells, and this activity was accounted for by PGE2. These studies define a novel autocrine/paracrine function of mediators produced by intestinal epithelial cells in the rapid induction of increased fluid secretion in response to intestinal infection with invasive bacteria. PMID- 9218507 TI - Neuronal cell death in Alzheimer's disease correlates with apoE uptake and intracellular Abeta stabilization. AB - The brains of individuals with Alzheimer's disease (AD) are characterized by extracellular deposition of beta-amyloid protein (Abeta), intracellular neurofibrillary tangles, and loss of neurons. To study molecular markers associated with dying cells in the AD brain, in situ DNA labeling techniques were used to visualize cells with DNA fragmentation. We observed that intracellular accumulation of apolipoprotein E (apoE) is correlated with the detection of intracellular Abeta-like immunoreactivity within the same cytoplasmic granules, suggesting that uptake of lipids may have stabilized the hydrophobic Abeta protein within the cell. These apoE-containing neurons also exhibit high expression of a cell surface receptor, gp330, which is known to bind apoE. Cells containing significant nuclear DNA fragmentation express the highest level of cell surface gp330. Extracellular deposition of Abeta is detected only upon neuronal cell death, initially as halos of Abeta immunoreactivity around individual dying neurons, and subsequently as Abeta plaques containing numerous neuronal cell ghosts. Based on our in situ analysis of nuclear DNA fragmentation, we conclude that neuronal cell death likely occurs before the extracellular deposition of Abeta in AD brains. PMID- 9218509 TI - Fragile-X: neuropsychological test performance, CGG triplet repeat lengths, and hippocampal volumes. AB - We compared cognitive performance and hippocampal volumes using magnetic resonance imaging (MRI) in adult fragile-X [fra(X)] males and females with either premutation (pM) or full mutation (fM) (n = 10 in all groups). Cognitive performance of fM males in the Wechsler Adult Intelligence Scale-Revised was worse than that of pM males, and the deficits in fM females were qualitatively similar, but less severe. In a visual memory test, both fM groups were impaired. In a list learning test, fM males were impaired in the learning phase and in delayed recognition. In a logical memory test, fM males and females were not significantly different from pM subjects. Hippocampal volumes normalized for intracranial or brain area did not significantly differ between fM and pM groups. However, positive correlations between left normalized hippocampal volumes and performance in many delayed memory tests observed in pM subjects were absent in fM subjects. Furthermore, in > 50% of the fM subjects, nonspecific changes, such as enlargement of ventricles and perivascular spaces, focal hyperintensities in temporal pole white matter, and/or subjectively assessed atypical appearance of hippocampal morphology, were observed in MRI. The data suggest minor abnormalities in temporal lobe structures in adult fra(X) subjects with fM. PMID- 9218508 TI - Ectopic induction of tendon and ligament in rats by growth and differentiation factors 5, 6, and 7, members of the TGF-beta gene family. AB - Little is known about the regulatory signals involved in tendon and ligament formation, and this lack of understanding has hindered attempts to develop biologically based therapies for tendon and ligament repair. Here we report that growth and differentiation factors (GDFs) 5, 6, and 7, members of the TGF-beta gene superfamily that are most related to the bone morphogenetic proteins, induce neotendon/ligament formation when implanted at ectopic sites in vivo. Analysis of tissue induced by GDF-5, 6, or 7, containing implants by currently available morphological and molecular criteria used to characterize tendon and ligament, adds further evidence to the idea that these GDFs act as signaling molecules during embryonic tendon/ligament formation. In addition, comparative in situ localizations of the GDF-5, 6, and 7 mRNAs suggest that these molecules are important regulatory components of synovial joint morphogenesis. PMID- 9218510 TI - Immunodominance of a low-affinity major histocompatibility complex-binding myelin basic protein epitope (residues 111-129) in HLA-DR4 (B1*0401) subjects is associated with a restricted T cell receptor repertoire. AB - The pathogenesis of multiple sclerosis (MS) is currently ascribed in part to a T cell-mediated process targeting myelin components. The T cell response to one candidate autoantigen, myelin basic protein (MBP), in the context of HLA-DR15Dw2, has been previously studied in detail. However, the characteristics of cellular immunity in the context of other MS-associated HLA-DR haplotypes are scarcely known. MBP-specific T cell lines (TCL) were generated from HLA-DR4 (B1*0401) positive MS subjects. Out of 275 MBP-specific TCL, 178 (64. 7%) specifically recognized region MBP(111-129), predominantly in the context of DRB1*0401. The major T cell epitope for MBP recognition corresponded to residues MBP(116-123). These TCL expressed disparate profiles of cytokine secretion and cytotoxicity. T cell receptor analysis, on the other hand, revealed a strikingly limited heterogeneity of rearrangements. In contrast to MBP(81-99), which binds with high affinity to HLA-DR15 and is recognized by a diverse T cell repertoire, MBP(111 129) binds weakly to DRB1*0401, suggesting that only high affinity T cell receptors might be able to efficiently engage such unstable MHC/peptide complexes, thus accounting for the T cell receptor restriction we observed. This study provides new insight about MBP recognition and proposes an alternative mechanism for immunodominance of self-antigen T cell epitopes in humans. PMID- 9218511 TI - Nitric oxide released from activated platelets inhibits platelet recruitment. AB - Vessel injury and thrombus formation are the cause of most ischemic coronary syndromes and, in this setting, activated platelets stimulate platelet recruitment to the growing thrombus. Recently, a constitutive nitric oxide synthase (NOS) has been identified in human platelets. To further define the capacity of platelets to produce nitric oxide (NO), as well as to study the role of this NO in platelet recruitment, we adapted a NO-selective microelectrode for use in a standard platelet aggregometer, thereby permitting simultaneous measurement of platelet aggregation and NO production. Treatment of platelets with the NO synthase inhibitor -NG-nitroarginine methyl ester (L-NAME), reduced NO production by 92+/-8% in response to 5 microM ADP compared to control but increased aggregation by only 15+/-2%. In contrast, L-NAME had a more pronounced effect on platelet recruitment as evidenced by a 35+/-5% increase in the extent of aggregation, a 33+/-3% decrease in cyclic GMP content, and a 31+/-5% increase in serotonin release from a second recruitable population of platelets added to stimulated platelets at the peak of NO production. To study platelet recruitment accurately, we developed an assay that monitors two platelet populations simultaneously. Nonbiotinylated platelets were incubated with L-NAME or vehicle and activated with ADP. At peak NO production, biotinylated platelets were added. As measured by three-color flow cytometry, there was a 56+/-11% increase in the number of P selectin- positive platelets in the nonbiotinylated population treated with L-NAME as compared to control. When biotinylated platelets were added to the L-NAME-treated nonbiotinylated population, the number of P selectin positive biotinylated plate-lets increased by 180+/-32% as compared to biotinylated platelets added to the control. In summary, stimulated platelets produce NO that modestly inhibits platelet activation but markedly inhibits additional platelet recruitment. These data suggest that platelet-derived NO may regulate platelet recruitment to a growing thrombus. PMID- 9218512 TI - Cytokine-dependent gp130 receptor subunit regulates human fetal pituitary adrenocorticotropin hormone and growth hormone secretion. AB - We have shown recently that leukemia inhibitory factor (LIF) and oncostatin M (OSM), two members of the gp130-dependent cytokine family, stimulate murine proopiomelanocortin (POMC) transcription and adrenocorticotropin hormone (ACTH) secretion. LIF and corticotropin-releasing hormone (CRH) also synergistically induced in vivo ACTH secretion in fetal nonhuman primates. To elucidate the role of the gp130-related cytokines in human pituitary hormone regulation, we tested expression of gp130-related cytokine receptors in human fetal pituitaries. Using RT-PCR, mRNA expression of receptors for LIF, IL-6, and CRH, and the gp130 subunit, were all detected in fetal pituitaries of 18- and 31-wk gestation. Recombinant human IL-6, LIF, and OSM treatments of primary human fetal pituitary cultures (16-31 wk) increased ACTH secretion by up to 48% (P < 0.05) using doses of 1 nM, and when fetal cultures were cotreated with CRH, ACTH was induced five- to sixfold as compared to CRH alone (three- to fourfold; P = 0.01). Incubation with gp130-specific antibody suppressed basal and cytokine-stimulated ACTH secretion (alone or with CRH) from human fetal cells. Human POMC promoter -879/+6 fused to the luciferase reporter gene and transfected into AtT-20 cells, was stimulated by LIF (7-fold), which also exerted strong (22-fold) synergy with CRH on POMC transcription. Growth hormone (GH) release from fetal cultures was modestly stimulated (15-31%, P < 0.05), while other anterior pituitary hormones were not altered by these cytokines. Thus, physiologic concentrations of the gp130-related cytokines have direct effects on ACTH and GH regulation in the human pituitary, indicating that gp130-dependent signals serve as a paracrine system controlling early human pituitary function. PMID- 9218513 TI - Induction of protective immunity after escherichia coli bladder infection in primates. Dependence of the globoside-specific P-fimbrial tip adhesin and its cognate receptor. AB - Clinical observations suggest that immune mechanisms affect etiology and course of recurrent cystitis. A primate infection model was used to show that primary bladder infection with a uropathogenic P-fimbriated strain (binding to globoside present in the bladder wall) protects against rechallenge with homologous as well as heterologous Escherichia coli strains for up to 5-6 mo. In contrast, mutant derivatives producing P-fimbriae either lacking the tip adhesin protein or carrying an adhesin for which no bladder receptor was present, were unable to induce protection, even though they generated bladder infections of similar duration as the wild type. Therefore, the protective effect mediated by the adhesin seemed to depend upon the presence of its cognate receptor. Since the wild strain also mediated protection against mutants that lacked the adhesin, our data suggest that the globoside-binding PapG adhesin acts as an adjuvant during infection to enhance a specific response against other bacterial antigens. In fact, the globoside-binding strain DS17, but not the mutant DS17-1, unable to bind to membrane-bound globoside, elicited a secretory IgA response to LPS in urine. These in vivo findings suggest that bacterial adhesin-ligand interactions may have signaling functions of importance for the immune response. PMID- 9218514 TI - The RHD gene is highly detectable in RhD-negative Japanese donors. AB - Recent molecular studies on the Rh blood group system have shown that the Rh locus of each haploid RhD-positive chromosome is composed of two structural genes: RHD and RHCE, whereas the locus is made of a single gene (RHCE) on each haploid RhD-negative chromosome. We analyzed the presence or absence of the RHD gene in 130 Japanese RhD-negative donors using the PCR method. The RhD-negative phenotypes consisted of 34 ccEe, 27 ccee, 17 ccEE, 26 Ccee, 19 CcEe, 1 CcEE, and 6 CCee. Among them, 36 (27.7%) donors demonstrated the presence of the RHD gene. Others showed gross or partial deletions of the RHD gene. These results were confirmed by Southern blot analysis. Additionally, the RHD gene detected in the RhD-negative donors seemed to be intact through sequencing of the RhD polypeptide cDNA and the promoter region of RHD gene. The phenotypes of these donors with the RHD gene were CC or Cc, but not cc. It suggested that there is some relationship between the RHD gene and the RhC phenotypes in RhD-negative individuals. In Caucasian RhD-negative individuals, the RHD gene has not been found outside of the report of Hyland et al. (Hyland, C.A., L.C. Wolter, and A. Saul. 1994. Blood. 84:321-324). The discrepant data on the RHD gene in RhD-negative donors between Japanese and Caucasians appear to be derived from the difference of the frequency of RhD-negative and RhC-positive phenotypes. Careful attention is necessary for clinicians in applying RhD genotyping to clinical medicine. PMID- 9218515 TI - Overexpression of the rat sarcoplasmic reticulum Ca2+ ATPase gene in the heart of transgenic mice accelerates calcium transients and cardiac relaxation. AB - The Ca2+ ATPase of the sarcoplasmic reticulum (SERCA2) plays a dominant role in lowering cytoplasmic calcium levels during cardiac relaxation and reduction of its activity has been linked to delayed diastolic relaxation in hypothyroid and failing hearts. To determine the contractile alterations resulting from increased SERCA2 expression, we generated transgenic mice overexpressing a rat SERCA2 transgene. Characterization of a heterozygous transgenic mouse line (CJ5) showed that the amount of SERCA2 mRNA and protein increased 2. 6-fold and 1.2-fold, respectively, relative to control mice. Determination of the relative synthesis rate of SERCA2 protein showed an 82% increase. The mRNA levels of some of the other genes involved in calcium handling, such as the ryanodine receptor and calsequestrin, remained unchanged, but the mRNA levels of phospholamban and Na+/Ca2+ exchanger increased 1.4-fold and 1.8-fold, respectively. The increase in phospholamban or Na+/Ca2+ exchanger mRNAs did not, however, result in changes in protein levels. Functional analysis of calcium handling and contractile parameters in isolated cardiac myocytes indicated that the intracellular calcium decline (t1/2) and myocyte relengthening (t1/2) were accelerated by 23 and 22%, respectively. In addition, the rate of myocyte shortening was also significantly faster. In isolated papillary muscle from SERCA2 transgenic mice, the time to half maximum postrest potentiation was significantly shorter than in negative littermates. Furthermore, cardiac function measured in vivo, demonstrated significantly accelerated contraction and relaxation in SERCA2 transgenic mice that were further augmented in both groups with isoproterenol administration. Similar results were obtained for the contractile performance of myocytes isolated from a separate line (CJ2) of homozygous SERCA2 transgenic mice. Our findings suggest, for the first time, that increased SERCA2 expression is feasible in vivo and results in enhanced calcium transients, myocardial contractility, and relaxation that may have further therapeutic implications. PMID- 9218516 TI - Vasoactive intestinal peptide prevents excitotoxic cell death in the murine developing brain. AB - Excitotoxic damage may be a critical factor in the formation of brain lesions associated with cerebral palsy. When injected at birth, the glutamatergic analog ibotenate induces mouse brain lesions that strikingly mimic human microgyria. When ibotenate is injected at postnatal day 5, it produces transcortical necrosis and white matter cysts that mimic human perinatal hypoxic-like lesions. Vasoactive intestinal peptide (VIP) has potent growth-related actions and neuroprotective properties that influence mitosis and neuronal survival in culture. The goal of this study was to assess the protective role of VIP against excitotoxic lesions induced by ibotenate in developing mouse brain. VIP cotreatment reduced ibotenate-induced microgyric-like cortical lesions and white matter cysts by up to 77 and 85%, respectively. VIP protective effects were reproduced by a peptide derived from activity-dependent neurotrophic factor (ADNF), a trophic factor released by VIP-stimulated astrocytes, and by stearyl norleucine VIP, a specific VIP agonist that does not activate adenylate cyclase. Neither forskolin, an adenylate cyclase activator, nor pituitary adenylate cyclase-activating peptide, provided VIP-like protection. VIP and neurotrophic analogs, acting through a cAMP-independent mechanism and inducing ADNF release, could represent new avenues in the understanding and prevention of human cerebral palsy. PMID- 9218517 TI - The insulinotropic potency of fatty acids is influenced profoundly by their chain length and degree of saturation. AB - Lowering of the elevated plasma FFA concentration in 18- 24-h fasted rats with nicotinic acid (NA) caused complete ablation of subsequent glucose-stimulated insulin secretion (GSIS). Although the effect of NA was reversed when the fasting level of total FFA was maintained by coinfusion of soybean oil or lard oil (plus heparin), the more saturated animal fat proved to be far more potent in enhancing GSIS. We therefore examined the influence of individual fatty acids on insulin secretion in the perfused rat pancreas. When present in the perfusion fluid at 0.5 mM (in the context of 1% albumin), the fold stimulation of insulin release from the fasted pancreas in response to 12.5 mM glucose was as follows: octanoate (C8:0), 3.4; linoleate (C18:2 cis/cis), 5.3; oleate (C18:1 cis), 9.4; palmitate (C16:0), 16. 2; and stearate (C18:0), 21.0. The equivalent value for palmitoleate (C16:1 cis) was 3.1. A cis--> trans switch of the double bond in the C16:1 and C18:1 fatty acids had only a modest, if any, impact on their potency. A similar profile emerged with regard to basal insulin secretion (3 mM glucose). When a subset of these fatty acids was tested in pancreases from fed animals, the same rank order of effectiveness at both basal and stimulatory levels of glucose was seen. The findings reaffirm the essentiality of an elevated plasma FFA concentration for GSIS in the fasted rat. They also show, however, that the insulinotropic effect of individual fatty acids spans a remarkably broad range, increasing and decreasing dramatically with chain length and degree of unsaturation, respectively. Thus, for any given level of glucose, insulin secretion will be influenced greatly not only by the combined concentration of all circulating (unbound) FFA, but also by the makeup of this FFA pool. Both factors will likely be important considerations in understanding the complex interplay between the nature of dietary fat and whole body insulin, glucose, and lipid dynamics. PMID- 9218518 TI - A novel pancreatic endocrine tumor suppressor gene locus on chromosome 3p with clinical prognostic implications. AB - The molecular pathogenesis of pancreatic endocrine tumors is largely unknown. Such tumors are more likely to develop in individuals with the von Hippel-Lindau (VHL) syndrome. We sought to determine whether allelic loss of the recently identified VHL tumor suppressor gene on chromosome 3p25-26 occurs in the more common sporadic forms of these tumors. Allelic loss on chromosome 3p was identified in 33% of 43 patients with endocrine tumors of the pancreas. The smallest common region of allelic loss, however, centered not at the VHL locus, but rather at 3p25, centromeric to VHL. Furthermore, no mutations of the VHL gene were identified in these tumors. Loss of alleles on chromosome 3p was associated with clinically malignant disease, whereas tumors with retained 3p alleles were more likely to be benign. Thus, the VHL gene does not appear to play a pathogenic role in the development of sporadic pancreatic endocrine tumors. Instead, a locus at chromosome 3p25 may harbor a novel pancreatic endocrine tumor suppressor gene, and allelic loss of this chromosomal region may serve as a molecular marker that helps distinguish benign from clinically malignant disease. PMID- 9218519 TI - Identification of functional endothelial protein C receptor in human plasma. AB - The endothelial cell protein C receptor (EPCR) binds protein C and facilitates activation by the thrombin-thrombomodulin complex. EPCR also binds activated protein C (APC) and inhibits APC anticoagulant activity. In this study, we detected a soluble form of EPCR in normal human plasma. Plasma EPCR appears to be approximately 43, 000 D, and circulates at approximately 100 ng/ml (98.4+/-27.8 ng/ml, n = 22). Plasma EPCR was purified from human citrated plasma using ion exchange, immunoaffinity, and protein C affinity chromatography. Flow cytometry experiments demonstrated that plasma EPCR bound APC with an affinity similar to that previously determined for recombinant soluble EPCR (Kdapp = 30 nM). Furthermore, plasma EPCR inhibited both protein C activation on an endothelial cell line and APC anticoagulant activity in a one-stage Factor Xa clotting assay. The physiological function of plasma EPCR is uncertain, but if the local concentrations are sufficiently high, particularly in disease states, the present data suggest that the soluble plasma EPCR could attenuate the membrane-bound EPCR augmentation of protein C activation and the anticoagulant function of APC. PMID- 9218520 TI - Cytosolic domain of the type I interleukin-1 receptor spontaneously recruits signaling molecules to activate a proinflammatory gene. AB - Immediate postreceptor events activated by IL-1-IL-1R interaction remain undefined. We have initiated studies to identify candidate signal transducers that associate with the cytosolic domain (cd) of the IL-1R. Immunocomplex kinase assays demonstrated an IL-1-activated myelin basic protein kinase activity that coprecipitated with the IL-1R from rat mesangial, mouse EL-4, and HeLa cells. Using glutathione-S-transferase (GST) fusion proteins, HeLa cell lysates next were assayed for kinases that associated with IL-1R cytoplasmic sequences. A GST IL-1R fusion protein containing the entire cd (amino acids 369-569; GST-IL-1Rcd) recruited a kinase activity in the absence and presence of IL-1 stimulation. In contrast, a GST-IL-1R membrane-proximal region mutant (amino acids 369-501; GST IL-1RcdDelta), which lacks COOH-terminal amino acid residues required for nuclear factor-kappaB activation, poorly phosphorylated MBP. In gel, kinase assays demonstrated 63-, 83-, and 100-kD kinases that specifically coprecipitated with the HeLa IL-1R and the GST-IL-1Rcd, but not GST-IL-1RcdDelta. 35S-labeled proteins, with Mrs identical to the kinase activities, stably associated with GST IL-1Rcd. Transient transfection assays of 293 cells were used to evaluate the functional significance of these findings. Simply increasing IL-1cd expression in 293 cells stimulated 5'-IL-6 flanking region-regulated CAT activity threefold above control, an effect blocked by the kinase inhibitors staurosporine and calphostin C. In summary, we have identified two previously unrecognized 63- and 83-kD kinases as well as a protein with an Mr similar to the recently cloned IL 1R-associated kinase, all of which associate spontaneously with the IL-1Rcd. Ectopic IL-1Rcd expression was sufficient to trigger cellular activation, suggesting that the extracellular domain of the intact receptor represses signal transduction until IL-1 is bound. Given that the IL-1Rcd signaling domain has been conserved in a functionally diverse group of transmembrane receptors, further characterization of this signaling process may define novel molecular mechanisms controlling cellular function and differentiation. PMID- 9218521 TI - Prevention of an increase in plasma cortisol during hypoglycemia preserves subsequent counterregulatory responses. AB - The aim of this study was to determine whether preventing increases in plasma cortisol during antecedent hypoglycemia preserves autonomic nervous system counterregulatory responses during subsequent hypoglycemia. Experiments were carried out on 15 (8 male/7 female) healthy, overnight-fasted subjects and 8 (4 male/4 female) age- and weight-matched patients with primary adrenocortical failure. 5 d before a study, patients had their usual glucocorticoid therapy replaced with a continuous subcutaneous infusion of cortisol programmed to produce normal daily circadian levels. Both groups underwent identical 2-d experiments. On day 1, insulin was infused at a rate of 1.5 mU/kg per min, and 2 h clamped hypoglycemia (53+/-2 mg/dl) was obtained during the morning and afternoon. The next morning, subjects underwent an additional 2-h hypoglycemic (53+/-2 mg/ dl) hyperinsulinemic clamp. In controls, day 2 steady state epinephrine, norepinephrine, pancreatic polypeptide, glucagon, growth hormone, and muscle sympathetic nerve activity were significantly blunted (P < 0.01) compared with day 1 hypoglycemia. In marked contrast, when increases of plasma cortisol were prevented in the patient group, day 2 neuroendocrine, muscle sympathetic nerve activity, hypoglycemic symptoms, and metabolic counterregulatory responses were equivalent with day 1 results. We conclude that (a) prevention of increases of cortisol during antecedent hypoglycemia preserves many critical autonomic nervous system counterregulatory responses to subsequent hypoglycemia; (b) hypoglycemia-induced increases in plasma cortisol levels are a major mechanism responsible for causing subsequent hypoglycemic counterregulatory failure; and (c) our results suggest that other mechanisms, apart from cortisol, do not play a major role in causing hypoglycemia-associated autonomic failure. PMID- 9218522 TI - Inhibition of constitutive nitric oxide synthase (NOS) by nitric oxide generated by inducible NOS after lipopolysaccharide administration provokes renal dysfunction in rats. AB - Excess NO generation plays a major role in the hypotension and systemic vasodilatation characteristic of sepsis. Yet the kidney response to sepsis is characterized by vasoconstriction resulting in renal dysfunction. We have examined the roles of inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS) on the renal effects of lipopolysaccharide administration by comparing the effects of specific iNOS inhibition, -N6-(1-iminoethyl)lysine (L-NIL), and 2,4 diamino6-hydroxy-pyrimidine vs. nonspecific NOS inhibitors (nitro- -arginine methylester). cGMP responses to carbamylcholine (CCh) (stimulated, basal) and sodium nitroprusside in isolated glomeruli were used as indices of eNOS and guanylate cyclase (GC) activity, respectively. LPS significantly decreased blood pressure and GFR (112+/-4 vs. 83+/-4 mmHg; 2.66+/-0.29 vs. 0. 96+/-0.22 ml/min, P < 0.05) and inhibited the cGMP response to CCh. GC activity was reciprocally increased. L-NIL and 2, 4-diamino-6-hydroxy-pyrimidine administration prevented the decrease in GFR (2.71+/-0.28 and 3.16+/-0.18 ml/min, respectively), restored the normal response to CCh, and GC activity was normalized. In vitro application of L-NIL also restored CCh responses in LPS glomeruli. Neuronal NOS inhibitors verified that CCh responses reflected eNOS activity. L-NAME, a nonspecific inhibitor, worsened GFR (0.41+/-0.15 ml/min), a reduction that was functional and not related to glomerular thrombosis, and eliminated the CCh response. No differences were observed in eNOS mRNA expression among the experimental groups. Selective iNOS inhibition prevents reductions in GFR, whereas nonselective inhibition of NOS further decreases GFR. These findings suggest that the decrease in GFR after LPS is due to local inhibition of eNOS by iNOS, possibly via NO autoinhibition. PMID- 9218523 TI - Alterations in skeletal muscle protein-tyrosine phosphatase activity and expression in insulin-resistant human obesity and diabetes. AB - Obese human subjects have increased protein-tyrosine phosphatase (PTPase) activity in adipose tissue that can dephosphorylate and inactivate the insulin receptor kinase. To extend these findings to skeletal muscle, we measured PTPase activity in the skeletal muscle particulate fraction and cytosol from a series of lean controls, insulin-resistant obese (body mass index > 30) nondiabetic subjects, and obese individuals with non-insulin-dependent diabetes. PTPase activities in subcellular fractions from the nondiabetic obese subjects were increased to 140-170% of the level in lean controls (P < 0.05). In contrast, PTPase activity in both fractions from the obese subjects with non-insulin dependent diabetes was significantly decreased to 39% of the level in controls (P < 0.05). By immunoblot analysis, leukocyte antigen related (LAR) and protein tyrosine phosphatase 1B had the greatest increase (threefold) in the particulate fraction from obese, nondiabetic subjects, and immunodepletion of this fraction using an affinity-purified antibody directed at the cytoplasmic domain of leukocyte antigen related normalized the PTPase activity when compared to the activity from control subjects. These findings provide further support for negative regulation of insulin action by specific PTPases in the pathogenesis of insulin resistance in human obesity, while other regulatory mechanisms may be operative in the diabetic state. PMID- 9218524 TI - Stimulation of suppressive T cell responses by human but not bacterial 60-kD heat shock protein in synovial fluid of patients with rheumatoid arthritis. AB - In several animal models of rheumatoid arthritis (RA), T cell responses to self 60-kD heat-shock protein 60 (hsp60) protect against the induction of arthritis. The nature of this suppressive T cell activity induced by self hsp60 is not clear. In the present study, T cell responses to human (self) hsp60 in RA in terms of type 1 (T1) and type 2 (T2) T cell activity were assessed. The results show that human and not bacterial hsp60-reactive synovial fluid (SF) T cells of patients with RA proliferate in the presence of the T2 cell growth factor IL-4. SF T cells stimulated with human hsp60 produced significantly lower amounts of IFN-gamma and higher amounts of IL-4 than SF T cells stimulated with bacterial hsp60 (P H(II) phase transition occurred from an osmotic pressure of 76.1 MPa for DOPE:DOPC (50:50) to 20 MPa in DOPE:DOPC:beta-sitosterol:CER (22.5:22.5:50:5) and 8 MPa in DOPE:DOPC:beta-sitosterol:CER (15:15:50:20). Experiments examining the effects of CER on the thermally-induced formation of the H(II) phase in fully hydrated mixtures and examining the influence of CER on the formation of the H(II) phase in DOPE:DOPC mixtures lacking beta-sitosterol suggested that CER facilitated the L(alpha) --> H(II) phase transition by effecting a decrease in bilayer hydration and by increased lateral packing pressures within the acyl domain of the bilayer. Taken in sum, these data indicate that the differential propensity of the rye and oat plasma membranes to undergo freeze-induced formation of the L(alpha) --> H(II) phase cannot be attributed to one lipid species. Rather, the propensity towards freeze-induced membrane destabilization is a consequence of the summation of physical characteristics of the membrane lipid components that included bilayer hydration, packing pressures within the hydrophobic domain of the membrane, the propensity of the lipid components to demix, and the relative proportions of the various lipid components. PMID- 9218554 TI - Grafted poly-(ethylene glycol) on lipid surfaces inhibits protein adsorption and cell adhesion. AB - Monolayers of dipalmitoyl-phosphatidylethanolamine (DPPE) mixing with various mole percentages of distearoyl-phosphatidylethanolamine (DSPE)-conjugated poly (ethylene glycol) (PEG m.w. 750-5000) were deposited on DPPE-coated glass surfaces by the Langmuir-Blodgett method. Increasing percentages of grafted PEG in these supported lipid surfaces increasingly inhibit the adsorption of bovine serum albumin (BSA), laminin, and fibronectin. Increasing percentages of grafted PEG also inhibit the adhesion of erythrocytes, lymphocytes, and macrophages to these supported lipid surfaces. The adsorption of proteins on lipid coated glass surfaces were assayed by the fluorescence of FITC-labelled proteins. Cell adhesion was measured mainly by microscopic counting. The concentration of PEG grafted lipids required for the inhibition of erythrocyte adhesion decreases with increasing molecular weight of the grafted PEG. The inhibitory effects are strongly dependent on the graft density of PEG at low concentrations, but weakly dependent on graft density at higher concentrations. For DSPE-PEG5000, the change of graft density dependency occurs approximately at the complete coverage of the lipid surface by the grafted polymer in the mushroom conformation (0.7 mol%), and the transition to partial brush conformation. The change-overs become less distinctive for grafted PEG of lower molecular weights, probably due to the failure of strictly mushroom and brush models of the polymer. The relative inhibitory efficiency is protein or cell dependent. The implication on the function of stealth liposomes is discussed. PMID- 9218555 TI - Exploring an antifungal target in the plasma membrane H(+)-ATPase of fungi. AB - The plasma membrane H(+)-ATPase is a promising new antifungal target that is readily probed with the sulfhydryl-reactive reagent omeprazole. Inhibition of the H(+)-ATPase by omeprazole is closely linked to cell killing, and it has been suggested that enzyme inhibition may result from a covalent interaction within the first two transmembrane segments (M1 and M2) (Monk et al. (1995) Biochim. Biophys. Acta 1239, 81-90). In this study, the molecular nature of this interaction was examined by screening a series of 26 well-characterized pma1 mutations residing in the first two transmembrane segments of the H(+)-ATPase from Saccharomyces cerevisiae. Only two pma1 mutants, A135G and G158D,G156C, were found to significantly decrease the sensitivity of cells for omeprazole. In contrast, enhanced sensitivity was observed at a number of positions, with D140C(A) and M128C producing the most significant increases in sensitivity. The introduction of cysteine at various locations within this region only marginally affected omeprazole sensitivity, suggesting that this region was not a direct site of covalent modification. Rather, its conformation influences omeprazole binding at some other locus. In order to determine the sidedness of the omeprazole interaction, a novel in vitro assay system was exploited that utilized liposomes co-reconstituted with the H(+)-ATPase and the light-driven proton pump bacteriorhodopsin. Omeprazole was found to completely inhibit proton transport by the H(+)-ATPase at 50 microM in this system. An asymmetrically-distributed chemical trap system involving glutathione was used to demonstrate that this inhibition appears localized to the extracellular portion of the enzyme. This work indicates that omeprazole can inhibit the H(+)-ATPase from its extracellular face, and this inhibition is influenced by changes in the M1, M2 region of the protein. PMID- 9218556 TI - The molecular area characteristics of the HIV-1 gp41-fusion peptide at the air/water interface. Effect of pH. AB - The putative fusion peptide of HIV-1 is a highly surface active substance. Relevant measurements with the Langmuir monolayer technique have been carried out for a broad range of the pH in the aqueous subphase. The data are processed towards a quantitative analysis of the partitioning equilibrium between the interfacial and aqueous moieties. Our results reveal a pronounced decrease of the surface area per peptide molecule upon monolayer compression. This phenomenon could be interpreted in terms of an orientational transition experienced by an alpha-helical peptide structure. The area requirements at any fixed lateral pressure pass through a distinct minimum at a pH of 5.5 (+/-0.5). Such an apparent isoelectric point was confirmed by isoelectric focusing of peptide aggregates. Accordingly a drastic drop of the pK-values of the two basic amino acid residues in comparison with an aqueous medium is indicated. It can be readily explained based on an inherent decrease of the effective dielectric constant. The observed low pH in favor of an enhanced surface affinity of the peptide may be a significant factor concerning its function as a fusion promoting agent. PMID- 9218557 TI - Ursodeoxycholate stabilizes phospholipid-rich membranes and mimics the effect of cholesterol: investigations on large unilamellar vesicles. AB - Ursodeoxycholate is used to treat primary biliary cirrhosis and is incorporated into hepatocyte plasma membranes. Its steroid nucleus binds to the apolar domain of the membrane, in a similar position to cholesterol. Therefore the question arises whether ursodeoxycholate has a similar effect on membrane structure and stability as cholesterol. Using differential scanning calorimetry the thermotropic behavior of egg phosphatidylcholine and dimyristoylphosphatidylcholine were studied after incubation with cholesterol or ursodeoxycholate. Large unilamellar vesicles were prepared with cholesterol contents of 0-50%. Following incubation of these vesicles with different amounts of ursodeoxycholate, vesicle stability in a gravitational field was investigated by measuring the phospholipid and cholesterol release. Vesicle size was studied by laser light scattering after incubation with cheno- and ursodeoxycholate, and the release of entrapped carboxyfluorescein was measured by means of fluorescence spectroscopy. Increasing cholesterol diminished the enthalpy of the phase transition in the membrane. Ursodeoxycholate decreased the enthalpy of the phase transition at even lower concentrations. Lipid release from vesicles in a high gravitational field diminished with increasing cholesterol content of the vesicles. Ursodeoxycholate had a comparable effect, which increased as the cholesterol content of the vesicles was decreased. Chenodeoxycholate damaged vesicles, whereas ursodeoxycholate did not. Cholesterol and ursodeoxycholate (below its critical micellar concentration) decreased the carboxyfluorescein release from vesicles induced by chenodeoxycholate. Thus like cholesterol, ursodeoxycholate is incorporated into phospholipid model membranes and reduces the change in enthalpy of the gel to liquid-crystalline phase transition. Like cholesterol ursodeoxycholate also maintains membrane stability and prevents membrane damage induced by mechanical and chemical stress. PMID- 9218558 TI - A predicted beta-sheet from class S components of staphylococcal gamma-hemolysin is essential for the secondary interaction of the class F component. AB - Site-directed mutagenesis was performed on genes encoding HlgA and HlgC, two of the three proteins expressed from the staphylococcal y-hemolysin locus, which originate two pore-forming toxins (HlgA + HlgB, HlgC + HlgB). As related proteins, HlgA and HlgC were found to bind first to cell membranes. Amino acid substitutions concerned residues that would predictably disrupt a 13 amino acid conserved beta-sheet of the Chou and Fasman secondary structure prediction. The mutation of a threonin into an aspartic acid residue from HlgA (T28D) and from HlgC (T30D) that would break this predicted N-terminal structure lowered dramatically the biological activities on purely lipidic vesicles, erythrocytes and polymorphonuclear cells. The change in secondary structure was confirmed by Fourier Transformed Infrared spectroscopy. The binding of mutated and native proteins at the same kind of sites onto polymorphonuclear cells was evidenced with flow cytometry and fluorescein-labelled anti-class S antibodies or wild type HlgA or HlgC. However, the subsequent binding of fluorescein-labelled HlgB to membrane-bound mutated HlgA or HlgC complexes was inhibited. In conclusion, the first binding of class S components is essential for the subsequent binding of class F components, and a predicted beta-sheet seems to be at least one of the functional domains involved. PMID- 9218559 TI - Uptake and export of citric acid by Aspergillus niger is reciprocally regulated by manganese ions. AB - The uptake as well as the export of citric acid by Aspergillus niger occur by active, deltapH-driven, H(+)-symport dependent systems. They are inhibited by nonmetabolizable tricarboxylic acid analogues and phthalic acid, and by several other mono-, di- and tribasic organic acids. However, citrate export could only be demonstrated in a mycelium cultivated under manganese-deficient growth conditions, whereas the uptake of citrate from the medium was only detectable upon precultivation of A. niger in a medium supplemented with Mn2+ ions. In addition, the uptake of citrate was dependent on the presence of Mn2+ ions in the assay, and inhibited by EDTA. This requirement for Mn2+ could also be partially fulfilled by Mg2+, Fe2+ or Zn2+, whereas Cu2+ ions inhibited citrate transport. The observed divergent effects of manganese ions on citrate uptake and citrate export may be a major reason for the well documented requirement for manganese deficiency of citric acid accumulation. PMID- 9218560 TI - Self-association of Band 3, the human erythrocyte anion exchanger, in detergent solution. AB - Dimeric Band 3 purified in n-dodecyl octaethyleneglycol (C12E8) underwent an irreversible, temperature-dependent association, resulting in a complex with a Stokes radius slightly larger than a native tetramer, before forming a higher molecular weight aggregate. Self-association occurred with a half-time of about 1 h at 37 degrees C but did not occur at 0 degrees C after several days. No change in the secondary structure of Band 3, as observed by circular dichroism, occurred during the association process. However, self-association of Band 3 was accompanied by loss of the stilbene disulfonate inhibitor binding site. No association or loss of inhibitor binding occurred with the dimeric membrane domain under similar incubation conditions. The membrane domain dimer was also stable over a wide range of pH (5.5-9.5) and buffer conditions, while Band 3 aggregated below pH 6.5. Inhibitors of anion transport, which stabilize the membrane domain, slowed the association. Band 3, depleted of phospholipids by extensive washing of resin-bound protein with detergent or, incubated with excess detergent, was more prone to aggregation. The membrane domain also showed some aggregation when depleted of lipids. Preparations could be stabilized by adding dimyristoylphosphatidylcholine (DMPC) prior to the 37 degrees C incubation. The effect of inhibitors and DMPC was additive, with a combination of 1 mM 4,4' dinitrostilbene-2,2'-disulfonate (DNDS) and 1:1 (wt/wt) DMPC:Band 3 stabilizing 90% of the protein to a 24-h incubation at 37 degrees C. The results suggest that self-association of Band 3 dimers is promoted by the cytoplasmic domain but results in alterations to the membrane domain involving the loss of essential phospholipids. Addition of phospholipid or inhibitors to Band 3 results in a stable preparation of the intact protein that may be suitable for crystallization studies. PMID- 9218561 TI - Outer membrane cytochromes of Shewanella putrefaciens MR-1: spectral analysis, and purification of the 83-kDa c-type cytochrome. AB - The metal-reducing bacterium Shewanella putrefaciens MR-1 is known to localize a majority of its membrane-bound cytochromes to its outer membrane when grown under anaerobic conditions. In this study, pyridine hemochrome spectra confirmed that these outer membrane cytochromes are c-type, and electrophoretic data demonstrated the presence of four distinct outer membrane cytochromes, with apparent molecular masses of 150, 83, 65, and 53 kDa. Fourth-order derivative analysis of 77 K spectra of the outer membrane revealed four spectrally distinct c-type hemes, with peaks at 545.4, 548.0, 550.6, and 552.6 nm. Outer membrane cytochromes in the reduced state were rapidly re-oxidized by oxidized iron and manganese, which have previously been shown to serve as electron acceptors for anaerobic respiration in this bacterium. The 83-kDa outer membrane cytochrome was purified and a specific polyclonal antibody was generated against this protein. Western blot analysis demonstrated that the vast majority of this protein was localized to the outer membrane and an intermediate density membrane fraction of similar composition. Its levels, but not its subcellular distribution, were somewhat influenced by the electron acceptor used to support anaerobic growth, with levels higher in fumarate-grown cells relative to iron(III)- or trimethylamine N-oxide-grown cells. Its specific content in cells grown under aerobic conditions was only 14% of that of fumarate-grown cells, suggesting that a switch to anaerobic conditions significantly increases the de novo synthesis of this outer membrane cytochrome. PMID- 9218562 TI - Changes in the expression of voltage-gated K+ currents during development of human megakaryocytic cells. AB - We distinguished four distinct groups of megakaryocytic cells on the basis of their voltage-gated membrane currents. One group of 32 cells (15%), exhibited an inward rectifying current and had a diameter of 12 +/- 3.5 microm (mean +/- S.D.). A large group of 85 cells (39%) exhibited only a 'leakage-like' current and had a diameter of 15.8 +/- 3.7 microm. The other two groups of cells exhibited voltage-gated outward currents. One group consisted of 43 'I-type' cells (19%), with a diameter of 22.3 +/- 3.4 microm, for which the maximal outward current occurred for a voltage step from -60 to either 0 or +20 mV. For the last group of 60 'M-type' cells (27%), which had a diameter of 26.7 +/- 2.9 microm, the maximal outward current occurred for a voltage step from -60 to +80 mV, the largest voltage step used. The currents recorded in this study, from megakaryocytes having 'leakage-like' currents and 'I-type' currents, were indistinguishable from the voltage-gated currents of the megakaryocytes from myelogenous leukemia patients, in which voltage-gated currents were suppressed (Kapural, L., O'Rourke, F., Feinstein, M.B. and Fein, A. (1995) Blood 86, 1043), suggesting that the megakaryocytes from the myelogenous leukemia patients are a dedifferentiated or less mature form of megakaryocyte. PMID- 9218563 TI - Biodistribution of liposomes containing synthetic galactose-terminated diacylglyceryl-poly(ethyleneglycol)s. AB - We describe the synthesis of biodegradable poly(ethyleneglycol)-coupled galactolipids in which the galactose moiety is separated from a diacylglyceride lipid anchor by poly(ethylene glycol) chains of 10, 20 or 40 oxyethylene residues (PEG10/20/40). These Gal-PEG lipids (Gal-PEG-Lip) were incorporated in the bilayer of liposomes. The surface exposure of the galactose was investigated by aggregation experiments with ricinus communis agglutinin 120. Only the liposomes containing the PEG10 galactolipid aggregated with the lectin. Therefore liposomes were prepared containing Gal-PEG10-Lip and a trace amount of [3H]cholesteryl oleyl ether with an average diameter of approximately 100 nm and injected intravenously into rats. The Gal-PEG10-Lip liposomes were cleared from plasma with a T1/2 of 0.3 h. Identically sized and composed control liposomes without the Gal-PEG10-Lip had a T1/2 of approximately 12 h. The rapid plasma elimination of the Gal-PEG10-Lip liposomes could be attributed entirely to increased uptake by the liver amounting to more than 90% of injected dose. Uptake by the spleen was decreased to less than 1% of injected dose. A single injection of N acetylgalactosamine 1 min prior to Gal-PEG-Lip liposome administration reduced the initial rate of plasma clearance to control levels. The increased liver uptake was almost entirely attributable to increased uptake by the Kupffer cells. Incorporation of PEG-DSPE in the Gal-PEG10-Lip liposomes only partially reversed the effect of the galactolipid with respect to liver and spleen uptake as well as intrahepatic distribution. These experiments demonstrate that liposome surface exposed galactose residues, even if attached at the distal end of a poly(ethyleneglycol) chain anchored in the liposomal bilayer are effectively recognized by the galactose particle receptor on the Kupffer cells but fail to achieve significant targeting to the asialoglycoprotein receptor on the hepatocytes. PMID- 9218564 TI - Phosphoinositide metabolism in hereditary ovalocytic red blood cell membranes. AB - Metabolic depletion of hereditary ovalocytes leads, similar to normal red cells, to decreased intracellular concentrations of ATP and GSH as well as degradation of the phosphoinositides to phosphatidylinositol and diacylglycerol. In contrast to normal red cells, however, loss of ATP does not induce any gross shape transformations; even after extensive depletion the ovalocytes retain their initial elongated stomatocytic character. The mechanical properties of hereditary ovalocytes are associated with a deletion of nine amino acid residues in band 3. Since the deletion appears to increase the stiffness of a normally flexible region of band 3, connecting the N-terminal cytoplasmic domain with the membrane spanning domain, our results indicate that shape changes require a flexible attachment of the cytoskeleton to the membrane-spanning band 3. The results also imply that metabolism of phosphoinositide cannot be the only determinant of cell shape, as suggested by the bilayer-couple hypothesis, but also other factors are involved in metabolically induced shape transformations. PMID- 9218565 TI - Concurrent or sequential delta and beta TCR gene rearrangement during thymocyte development: individual thymi follow distinct pathways. AB - Analysis of TCR rearrangement profiles of well-defined thymocyte populations in a number of individual thymi provides evidence for a new pathway of lineage commitment. In all of the thymi analyzed, alphabeta thymocytes have rearrangements in the delta locus that are enhanced for out-of-frame rearrangements. Thus, not only did alphabeta thymocytes pass through a stage in differentiation that included delta rearrangement, but they also constitute a population that was relatively unsuccessful at these rearrangements. Interestingly, in some thymi, gammadelta thymocytes have out-of-frame beta rearrangements. This represents a novel pathway in which delta and beta rearrangement happens concurrently. In this pathway, selection favors whichever gene is in frame, thus improving the chances of generating useful T cells. In other thymi, gammadelta cells showed no obvious beta gene rearrangement, indicating a sequential rearrangement. Thus, alphabeta/gammadelta lineage commitment can follow at least two distinct pathways in different individuals. PMID- 9218566 TI - Dexamethasone potently enhances phorbol ester-induced IL-1beta gene expression and nuclear factor NF-kappaB activation. AB - The synthetic glucocorticoid dexamethasone, an immunosuppressive and anti inflammatory agent, was investigated for its effect on PMA-mediated expression of the inflammatory cytokine IL-1beta in the human monocytic leukemic cell line THP 1. PMA alone induced the production of low levels of IL-1beta in THP-1 cells, whereas dexamethasone alone had no effect. However, dexamethasone potently enhanced PMA-mediated IL-1beta production. Using a selective and potent inhibitor of protein kinase C, we found that synergistic interaction between PMA and dexamethasone requires protein kinase C activation. PMA has been known to activate nuclear factor NF-kappaB in THP-1 cells. Using an oligonucleotide probe corresponding to an NF-kappaB DNA-binding motif of the IL-1beta gene promoter in gel electrophoresis mobility shift assays, we demonstrated that PMA-induced NF kappaB activation was greatly potentiated by dexamethasone. Our results indicate that glucocorticoids can be positive regulators of inflammatory cytokine gene expression during monocytic cell differentiation. PMID- 9218567 TI - Reconstitution of the extrathymic intestinal T cell compartment in the absence of irradiation. AB - Extrathymic development of intestinal intraepithelial lymphocytes was studied using a reconstitution model that does not require irradiation. WBB6F1/J Kit(W)/Kit(W-v) mice were reconstituted with normal fetal liver cells. In this system, reduced c-Kit activity in host hemopoietic progenitors imparts normal precursors with a growth advantage and, thus, chimerism can be established without irradiation. In control mice, TCR gammadelta and TCR alphabeta intraepithelial lymphocytes (IEL) developed efficiently from fetal liver cells, with a predominance of TCR alphabeta over TCR gammadelta IEL. In contrast, development in reconstituted thymectomized mice was heavily skewed toward TCR gammadelta IEL generation. In thymectomized mice, development of CD4+ 8- and CD4+ 8+ TCR alphabeta IEL did not occur, while TCR alphabeta CD8 alphabeta development was nearly absent. The results indicated that without irradiation the majority of TCR alphabeta IEL were thymus dependent, whereas TCR gammadelta IEL developed extrathymically. Thus, the discrepancies observed between different models of athymic development may be explained by the induction of T cell development as a result of irradiation. PMID- 9218568 TI - Characterization of transport of newly assembled, T cell-stimulatory MHC class II peptide complexes from MHC class II compartments to the cell surface. AB - In human B cells MHC class II molecules acquire antigenic peptides in lysosome related compartments called the MHC class II compartments (MIIC). How assembled complexes, capable of activating T cells, then reach the cell surface has not been fully resolved. We have used selective ablation of early and recycling endosomes to determine whether newly peptide-loaded class II molecules require functional recycling endosomes to exit to the cell surface. Cellular compartments accessed by transferrin-horseradish peroxidase conjugates were functionally inactivated by cross-linking with diaminobenzidine and hydrogen peroxide. Cells with ablated endosomal compartments were unable to recycle transferrin to the cell surface and could not deliver exogenous Ag for processing and presentation to T cells. In contrast, cells that had taken up Ag and assembled intracellular class II-peptide complexes before endosome ablation were still able to deliver class II-peptide complexes to the cell surface and stimulate T cell proliferation. This delivery was abolished in the presence of brefeldin A. These data show that the parts of the endocytic apparatus necessary for Ag delivery to the MIIC are not required for functional class II-peptide complexes to reach the cell surface. Moreover, the brefeldin A sensitivity of this final step in the class II molecule biosynthetic pathway suggests a vesicular intermediate for transport between the MIIC and the plasma membrane. PMID- 9218569 TI - The generation of MHC class I-associated peptides is only partially inhibited by proteasome inhibitors: involvement of nonproteasomal cytosolic proteases in antigen processing? AB - The proteasome is believed to participate in the generation of a large percentage of peptide ligands for MHC class I molecules. This conclusion is based largely on the activities of peptidyl aldehydes that block proteasome activity. We tested the ability of a panel of proteasome inhibitors to affect the generation of MHC class I binding peptides in mouse L929 cells. Included in the panel are peptidyl aldehydes and a microbial product, lactacystin, that blocks proteasome activity in a distinct and more specific manner. Contrary to expectations, proteasome inhibitors failed to block the generation of a large portion of high affinity peptides as inferred by measuring cell surface expression of newly synthesized MHC class I molecules. These findings were confirmed by examining the effects of the inhibitors on the presentation of individual antigenic determinants from endogenously synthesized or exogenously delivered influenza virus proteins. Presentation of peptides derived from exogenous basic polymerase 1, endogenous basic polymerase 1, and nonstructural-1 proteins was decreased by inhibitors in a manner consistent with proteasomal involvement. Presentation of peptides derived from endogenous nucleoprotein was not significantly affected by the proteasome inhibitors, while presentation of exogenous hemagglutinin and nucleoprotein was enhanced by the proteasome inhibitors. These data are consistent with the involvement of both proteasomes and nonproteasomal cytosolic proteases in the generation of a significant portion of MHC class I binding peptides. PMID- 9218570 TI - Dendritic cell subtypes in mouse lymphoid organs: cross-correlation of surface markers, changes with incubation, and differences among thymus, spleen, and lymph nodes. AB - Freshly isolated, mature dendritic cells (DC) from mouse lymphoid organs were analyzed by immunofluorescent labeling and flow cytometry to determine the number of discrete subpopulations and to assess possible lineage markers. The permanence of surface markers was then determined by overnight culture of the DC. Three DC subtypes were discerned, CD8alpha- DEC-205-, CD8alpha+ DEC-205+, and CD8alpha- DEC-205+, with different tissue distributions. The majority of DC expressed high levels of class II MHC, expressed CD11c, and expressed the costimulator molecules CD80, CD86, and CD40; CD80 and CD40 were further up-regulated on culture. DC also expressed low levels of L-selectin that were up-regulated on culture. Thymus contained predominantly CD8alpha+ DEC205+ CD11b- DC, resembling a major subpopulation of DC in other tissues but unique in expressing BP-1. Spleen contained predominantly two DC populations in equal proportions: one CD8alpha+ DEC-205+ CD11b- as in the thymus, and the other CD8alpha- DEC-205- CD11b+. Lymph nodes contained the same two DC populations as in spleen, but in addition a third population of CD8alpha- DEC-205+ CD11b- DC. The CD8alpha expression of splenic DC subpopulations did not change on culture. Although DEC-205 was up-regulated on culture so all DC became positive, the difference in the level between subpopulations was maintained. However, CD11b was up-regulated on culture, so all subpopulations became positive and finally expressed equivalent levels. Some aspects of this complex, but discrete, pattern of surface marker expression can be correlated with differences in lineage origin and functional activity of the DC. PMID- 9218571 TI - Melatonin enhances IL-2, IL-6, and IFN-gamma production by human circulating CD4+ cells: a possible nuclear receptor-mediated mechanism involving T helper type 1 lymphocytes and monocytes. AB - This paper shows that melatonin is able to activate human Th1 lymphocytes by increasing the production of IL-2 and IFN-gamma in vitro. Th2 cells appear not to be affected by melatonin, since IL-4, which is mostly produced by Th2 cells, is not modified by the hormone. Melatonin also enhances IL-6 production by PBMCs. The activation by melatonin of IL-6 production is apparently related to the presence of monocytes, rather than to Th2 cells, in the cell preparation, since PBMCs depleted of monocytes (CD14+ cells) were not activated. Activation of PBMCs by melatonin was dependent on the dose and, measured by cytokine production, was observed only when cells were either not activated or only slightly activated by low concentrations of PHA, or when cell activation was achieved by incubating the cells with previously irradiated cells. Using a different approach to identify what type of cells among the PBMC subsets was activated by melatonin, the expression of CD69, a marker of cell activation, was studied. Melatonin increased the percentage of cells expressing the CD69 Ag in CD4+ but not in CD8+ cells. We have also achieved enhanced production of IL-2 and IL-6 using CGP 52608, a specific ligand of the putative nuclear melatonin receptor RZR/ROR, raising the possibility of direct effects of melatonin on gene regulation in both Th1 cells and monocytes. The results suggest that melatonin may be involved in the regulation of human immune functions by modulating the activity of Th1 cells and monocytes via nuclear receptor-mediated transcriptional control. PMID- 9218572 TI - The role of protein kinase C in CD8+ T lymphocyte effector responses. AB - Rare CD8+ T cells present in beta2microglobulin-deficient (beta2m-/-) mice reject allogeneic tumors but not syngeneic wild-type tumors. The lack of syngeneic tumor rejection in vivo is correlated with a partial response of beta2m-/- CD8+ cell lines to syngeneic tumor cells in vitro. This partial response is characterized by perforin/granzyme-mediated cytolytic activity in the absence of cytokine secretion or proliferation. Allogeneic tumors induce cytolysis, cytokine secretion, and proliferation. Cytokine secretion may therefore be an important effector mechanism for tumor rejection by CD8+ T cells. To determine the missing signaling events needed for cytokine secretion as well as the events inducing the isolated cytotoxic response, we attempted to restore cytokine secretion of beta2m /- CD8+ cells to syngeneic MHC class I. Phorbol ester and syngeneic tumor cells acted synergistically to induce full responsiveness of beta2m-/- CD8+ cells. However, this synergistic induction of cytokine secretion used a different pathway than that induced by alloantigen. Protein kinase C (PKC) inhibitor prevented the syngeneic class I plus PMA-induced cytokine secretion, but not allo class I-induced cytokine secretion. In contrast to the PKC independent alloantigen-induced cytokine secretion, cytolysis of both allogeneic and syngeneic targets was PKC dependent. The differential dependence of effector functions on PKC activation was also found in beta2m+/+ CD8+ T cells. Thus, two distinct signaling pathways (PKC dependent and PKC independent) may ultimately converge to induce cytokine secretion in CD8+ cells. The TCR engagement-initiated pathway is PKC independent, whereas the phorbol ester-activated pathway is PKC dependent. PMID- 9218573 TI - Inflammatory cytokines enhance the in vivo clonal expansion and differentiation of antigen-activated CD4+ T cells. AB - Despite the wealth of information on the signals required for T cell activation in vitro, the signals required for the generation of functional Th cells in vivo are poorly understood. We addressed this by directly tracking the behavior of adoptively transferred CD4+ TCR transgenic T cells following Ag administration in vivo. Injection of soluble Ag induced a transient accumulation of Ag-specific T cells in lymphoid tissue. If bacterial LPS was present during this period, enhanced numbers of Ag-specific T cells accumulated, migrated into B cell-rich follicles, and provided help for Ab production. The ability of LPS to enhance the accumulation and follicular migration of Ag-activated T cells was mimicked by the proinflammatory cytokines, TNF-alpha and IL-1, and the capacity of LPS to promote the generation of IFN-gamma-secreting T cells, which provide help for IgG2a production, was mimicked by IL-12. Thus, the in vivo generation of functional Th cells can arise from Ag-dependent clonal expansion in the context of inflammatory cytokines. PMID- 9218574 TI - Targeting of natural killer cells to mammary carcinoma via naturally occurring tumor cell-bound iC3b and beta-glucan-primed CR3 (CD11b/CD18). AB - Previous reports have suggested that malignant cells frequently generate a humoral immune response that is ineffective in tumor destruction. Despite coating tumors with IgM and IgG that activate the C system via the classical pathway, normal membrane regulators of C (e.g., membrane cofactor protein and CD59) prevent cytotoxicity. Moreover, C3 deposition on tumors does not result in cytotoxic recognition by phagocytes or NK cells bearing C3 receptors capable of mediating destruction of C3-opsonized bacteria or yeast. The current investigation showed that freshly excised mammary tumors bore IgM, IgG, and C3 detectable by flow cytometry. Normal sera contained natural IgM and IgG Abs reactive with breast tumor cell lines, and IgG Ab titers were increased in patients with breast cancer. Breast tumor cell lines incubated in normal serum from AB+ individuals activated the classical, but not the alternative, pathway of C and became coated with C3. Despite exhibiting membrane-bound C3, serum opsonized breast tumor cell lines were not killed by CR3 (CD11b/CD18)-bearing NK cells. Priming of NK cell CR3 with small soluble yeast beta-glucan polysaccharides enabled CR3-dependent killing of these same C3-bearing tumor cell lines. Tests of mammary carcinoma cells from freshly excised tumors demonstrated that they also bore sufficient amounts of opsonic C3 for cytotoxic recognition by NK cells bearing polysaccharide-primed CR3, whereas they were largely resistant to NK cells bearing unprimed CR3. This study demonstrates the potential utility of using naturally occurring opsonic C3 on tumor cells for specific immunotherapeutic targeting by NK cells and phagocytes bearing polysaccharide primed CR3. PMID- 9218575 TI - Cytokine-deficient CD8+ Tc1 cells induced by IL-4: retained inflammation and perforin and Fas cytotoxicity but compromised long term killing of tumor cells. AB - After antigenic stimulation, naive CD8+ T cells differentiate into cytotoxic Tc1 cells secreting the cytokines IL-2, IFN-gamma, and TNF, which aid their proliferation and effector functions. We have previously shown that IL-4 acts directly on differentiated Tc1 cells to impair subsequent Con A-induced IL-2 production. As IL-4 may be produced in the vicinity of Tc1 cells during normal immune responses, we have further analyzed the short and long term functions of IL-4-treated Tc1 cells. We now show that these cells also have a defect in the synthesis of IFN-gamma, TNF, and IL-10 in response to antigenic stimulation. IL-2 synthesis was the most sensitive, as stimulation of IL-4-treated Tc1 cells with higher numbers of APCs partially restored IFN-gamma, TNF, and IL-10, but not IL 2, synthesis. Injection of allo-specific Tc1 cells into mice expressing the target Ag revealed reduced cytokine synthesis in vivo by IL-4-treated Tc1 cells. Loss of cytokine synthesis did not impair the short term effector functions of Tc1 cells, as they induced adoptively transferred delayed type hypersensitivity in recipient mice and retained both perforin- and Fas-dependent cytolytic mechanisms in vitro. Long term coculture of tumor targets and tumor-specific Tc1 cells indicated that normal Tc1 cells proliferated and killed tumor cells, whereas IL-4-treated Tc1 cells failed to proliferate and hence were unable to curtail the proliferation of tumor cells. These results suggest that IL-4 synthesis in vivo would not affect immediate effector functions of differentiated Tc1 cells, but would compromise immunity by reducing their long term functional capability. PMID- 9218576 TI - The 25-kDa soluble CD23 activates type III constitutive nitric oxide-synthase activity via CD11b and CD11c expressed by human monocytes. AB - CD23, a low-affinity receptor for IgE, was recently shown to bind to CD11b and CD11c molecules on human monocytes. The 25-kDa soluble fragment of CD23 (sCD23), was tested for its capacity to elicit various signaling pathways in human monocytes. sCD23 was found to trigger an early increase in cGMP accumulation, through the generation of nitric oxide. This was a result of activating the L arginine pathway, since the sCD23-mediated effect was inhibited in the presence of substituted nonmetabolizable L-arginine analogues. In addition, the increase in cGMP was suppressed by calcium chelators and inhibitors of the calcium/calmodulin complex, suggesting the involvement of a constitutive, calcium dependent nitric oxide synthase (NOS). Indeed, the presence of an endothelial constitutive type III NOS mRNA was detected in nonactivated human monocytes, and the corresponding protein has been detected by flow cytometry. Moreover, sCD23 was shown to induce a calcium influx in monocytes, in accordance with an activation of a constitutive NOS through a transient increase in [Ca2+]i. As expected, these events were mimicked by mAbs against CD11b and CD11c, the macrophage receptors for CD23. In addition to these early events, sCD23 and the agonistic anti-CD11b and CD11c mAbs, which all trigger the release of proinflammatory mediators such as TNF-alpha, were shown to act through an NO dependent process. PMID- 9218577 TI - Compromised allograft rejection response in transgenic mice expressing antisense sequences to retinoic acid receptor beta2. AB - Transgenic mice expressing antisense sequences to retinoic acid receptor beta2 (AS-RARbeta2 mice) were generated. The transgene was expressed in T cells, macrophages, and non-T spleen cells. These AS-RARbeta2 mice had four- to sevenfold higher endogenous RARbeta2 messages in their macrophages, comparing with their nontransgenic littermates, but the RARbeta protein level was significantly lower in these cells. This suggests that the antisense message interferes with RARbeta2 translation, and there is a feedback control of the RARbeta2 mRNA level in the macrophages. The endogenous RARbeta2 message was not detectable in T and B cells of either the transgenic or nontransgenic mice. These mice appeared to be healthy upon visual inspection. When transplanted with allogenic cardiac grafts heterotopically, the mice showed significantly compromised rejection response. These mice had a decrease of the macrophage population in spleen, using the nontransgenic littermates as references. The generation of alloantigen-specific T cell cytotoxicity was decreased in these AS RARbeta2 mice. These results suggest that macrophage development and T cell function are affected by antisense expression of RARbeta2, and that anomaly in these cells might be contributing factors to the compromised immune response observed in the AS-RARbeta2 mice. PMID- 9218578 TI - Activation of human dendritic cells following infection with Mycobacterium tuberculosis. AB - Dendritic cells (DC) play an essential role in the initiation of primary T cell responses to foreign Ag. It is likely that these potent APC are critical in the initiation of immune responses to pathogens, such as bacteria or parasites. However, little is known about the interaction of these important APC with pathogens. To address this issue, the interaction of the bacterium Mycobacterium tuberculosis with human DC was studied. DC generated from human peripheral blood by short term culture in medium containing recombinant human cytokines granulocyte-macrophage-CSF and IL-4 were capable of phagocytosing M. tuberculosis. Infection of DC with live M. tuberculosis bacilli resulted in increased APC surface expression of the costimulatory molecules CD54, CD40, and B7.1, as well as MHC class I molecules. In addition, infected DC secreted elevated levels of inflammatory cytokines, including TNF-alpha, IL-1, and IL-12. M. tuberculosis-infected human monocytes also secreted inflammatory cytokines, but exhibited no enhancement of costimulatory or MHC class I molecule expression. These data indicate that infection with M. tuberculosis results in the direct activation and maturation of these DC. In vivo, such activation may facilitate migration to the lymph nodes, and enhance presentation of Ag to T cells, thereby facilitating the induction of the immune response against this pathogen. PMID- 9218579 TI - Induction of tolerance to human gamma-globulin in FcR gamma- and Fc gammaRII deficient mice. AB - FcR gamma-deficient mice were used to examine the role of Fc gamma receptors in the induction of peripheral tolerance to human gamma-globulin (HGG). FcR gamma deficient mice injected with HGG in adjuvant demonstrated a CD4+ T cell response to in vitro challenge with HGG, as assayed by proliferation, cytokine secretion, and Ag-specific help for B cell Ab production. In vitro kinetics of Ag-specific proliferation were similar in both conventional and knockout mice. Peripheral tolerance could be established in these mice with a single dose of deaggregated protein, despite the lack of functional Fc gammaRI, the high affinity receptor for monomeric IgG. Establishment of unresponsiveness was observed at both the T and B cell levels. T cell tolerance was manifested in the reduction of T cell helper function and Ag-induced release of Th1- and Th2-like cytokines, as well as decreased proliferation to Ag-specific stimulation. B cell tolerance was demonstrated in knockout and normal mice by failure to detect HGG-specific Ab production using an immunization protocol for Ab production that bypasses the need for Ag-specific T cells. These results demonstrate that induction of tolerance in CD4+ cells to HGG does not require transduction of a signal through Fc gammaRI. Furthermore, the ability to induce tolerance to HGG in B cells in Fc gammaRII-deficient mice suggests that down-regulation of Ag-specific B cells through Fc gammaRII is not the mechanism by which B cell tolerance is induced. However, Fc gammaRII plays a role in regulating the immune response since the Ab response to immunogenic HGG in Fc gammaRII-deficient mice is markedly enhanced. PMID- 9218580 TI - Migration and activation of antigen-specific CD8+ T cells upon in vivo stimulation with allogeneic tumor. AB - The response of CD8+ T cells to allogeneic tumor was studied by adoptive transfer of cells from TCR transgenic 2C mice specific for Ld alloantigen. Transferred cells were monitored during the course of a response to i.p. challenge with live P815 (H-2d) using the 1B2 mAb specific for the 2C TCR. Tumor was present in the draining LN and spleen within 3 to 4 days of challenge. The first changes in 1B2+ cells occurred in the spleen on day 4; VLA-4 expression increased in an Ag specific manner and L-selectin expression decreased in an Ag-nonspecific manner. The number of 1B2+ cells in the spleen declined over days 4 to 6. The first detectable increase in CD25 expression and blast transformation was in the peritoneal cavity beginning days 5 and 6. Clonal expansion was largely limited to this site and was maximal on day 8. As expansion occurred in the peritoneal cavity; the number of 1B2+ cells in the draining LN and spleen also increased. These cells had an activated phenotype (CD44(high), VLA-4(high)) but most did not express CD25 and were not blasts. These results suggest that initial Ag recognition in the spleen results in altered expression of adhesion receptors so that cells gain access to the peritoneal cavity where they undergo clonal expansion and differentiation. Following the response, 1B2+ cells decline in number but a memory population (CD44(high), L-selectin(high and low)) persists for long times in the spleen and LN. PMID- 9218581 TI - Type 1 versus type 2 cytokine release by Vbeta T cell subpopulations determines in vivo antitumor reactivity: IL-10 mediates a suppressive role. AB - We have previously reported that CD8+ tumor-draining lymph node (TDLN) cells activated with anti-CD3 and IL-2-mediated tumor regression in adoptive immunotherapy. In this study, we examined the TCR Vbeta repertoire usage of TDLN cells with respect to cytokine release profiles and therapeutic efficacy in vivo. The majority of the whole population of TDLN cells after activation with anti-CD3 were composed of Vbeta3+, -5+, -7+, -8+, and -11+ cells. Enrichment of Vbeta subsets of TDLN cells by in vitro activation with anti-Vbeta mAb revealed Vbeta8+ cells released high amounts of IFN-gamma and granulocyte/macrophage-CSF (GM-CSF) with minimal amounts of IL-10 in response to tumor and mediated tumor regression in vivo. In contrast, enriched populations of Vbeta5+, Vbeta7+, and Vbeta11+ cells released low amounts of IFN-gamma and GM-CSF with high levels of IL-10 and had no in vivo antitumor reactivity. In vitro depletion of specific Vbeta subsets from the whole TDLN pool confirmed that the profile of cytokines released correlated with in vivo antitumor function. Therapeutic efficacy mediated by TDLN cells required the release of IFN-gamma and GM-CSF since in vivo neutralization of both cytokines inhibited tumor regression. The administration of anti-IL-10 mAb abrogated the suppressed antitumor response manifested by adoptively transferred TDLN cells, which elaborated increased levels of IL-10. Our study documents that type 1 cytokine release (i.e., IFN-gamma and GM-CSF) promotes in vivo tumor Ag recognition, in contrast to type 2 release (i.e., IL-10), which suppresses this interaction, and discriminates the functional activity of Vbeta subpopulations of effector cells. PMID- 9218582 TI - Increased lymphocyte apoptosis in Fas ligand transgenic mice. AB - Fas ligand (fasL) transgenic (Tg) mice were produced by introducing a murine fasL cDNA under the control of a TCR beta-chain enhancer minigene. Higher levels of fasL expression with increased biologic activity were observed in the Tg mice compared with non-Tg mice. Numbers of CD4+ CD8+ T cells in the thymus and T cells in the lymph node and spleen were lower in the fasL Tg mice compared with the non Tg mice. This is consistent with a reduction in the size of the T cell areas in fasL Tg mice compared with non-Tg mice. Conversely, in fasL Tg mice, there was an increase in the number and size of apoptotic foci associated with phagocytic cells, as determined by in vivo TUNEL (TdT-mediated dUTP nick-end labeling) staining. Stimulation of non-Tg mice in vivo with anti-CD3 Ab for 3 days resulted in greatly increased apoptosis of CD4+ CD8+ thymocytes and lymph node T cells. Surviving thymocytes and T cells of lymph node and spleen expressed Fas at low levels. After similar stimulation of fasL Tg mice, however, a discreet population of surviving cells expressed high levels of Fas, indicating that a novel population of Fas apoptosis-resistant cells develops in these mice. These results indicate that high levels of fasL can result in both increased Fas-mediated apoptosis and the development of T cells that express high levels of Fas, but are resistant to Fas-mediated apoptosis. PMID- 9218583 TI - Adenoviral gene delivery elicits distinct pulmonary-associated T helper cell responses to the vector and to its transgene. AB - Replication-deficient adenovirus (Ad) vectors are effective to specifically target the respiratory epithelium for either corrective gene therapy such as cystic fibrosis or for mucosal immunization. As a consequence of transducing the lower respiratory tract with an E1/E3 deleted Ad5 vector, host responses have been characterized by the duration of transgene expression and by the induction of CTL responses. However, limited emphasis has been devoted to understanding the contribution of CD4+ T cell responses to the Ad vector. Both CD4+ and CD8+ T cells migrate into the lung following sequential intratracheal Ad5 transgene instillations. Isolated CD3+ T lymphocytes from the lungs were predominantly of the Th2 type, and after cell sorting, the IL-4-producing T cells were largely CD4+, while IFN-gamma expression was associated with both CD4+ and CD8+ T cells. Ab responses to the Ad5 vector and to the expressed transgene beta-galactosidase (beta gal) revealed elevated bronchial and serum IgA and IgG Abs with low neutralization titers. Analysis of serum IgG subclass responses showed IgG1 and IgG2b with lower IgG2a Abs to Ad5 and IgG2a and IgG2b Ab responses to beta gal. Ad5-specifc CD4+ T cells produced both Th1 (IFN-gamma and IL-2)- and Th2 (IL-4, IL-5, IL-6)-type cytokines, while beta gal-specific CD4+ T cells secreted IFN gamma and IL-6. This study provides direct evidence for the concomitant induction of Th2- with Th1-type responses in both the pulmonary systemic and mucosal immune compartments to the Ad5 vector as well as a Th1-dominant response to the transgene. PMID- 9218584 TI - Fetal liver contains committed NK progenitors, but is not a site for development of CD34+ cells into T cells. AB - The presence of T and NK cells in the human fetal liver and the fact that fetal liver hemopoietic progenitor cells develop into T and NK cells suggest a role for the fetal liver compartment in T and NK cell development. In this work, we show that the capacity of fetal liver progenitors to develop into T cells, in a human/mouse fetal thymic organ culture system, is restricted to an immature subset of CD34+ CD38- cells. No T cell-committed precursors are contained within the more differentiated CD34+ CD38+ population. This conclusion is supported by the observations that no TCR-delta gene rearrangements and no pre-TCR-alpha expression can be detected in this population. However, NK cells were derived from CD34+ CD38- and CD34+ CD38+ fetal liver cells cultured in the presence of IL 15, IL-7, and Flt-3 ligand. Eighty to ninety percent of cells arising from the CD34+ CD38+ population expressed the NK cell-associated markers CD56, CD16, CD94, and NKR-P1A. Several subpopulations of NK cell precursors were identified by differential expression of these receptors. Based on the detection of populations with a similar antigenic profile in freshly isolated fetal liver cells, we propose a model of NK cell differentiation. Collectively, our findings suggest that CD34+ cells differentiate into NK cells, but not into mature T cells, in the human fetal liver. PMID- 9218585 TI - Different modes of peptide interaction enable HLA-DQ and HLA-DR molecules to bind diverse peptide repertoires. AB - The role of HLA-DQ molecules in Ag presentation has, thus far, remained elusive. Here we report that two DQ allotypes, DQ7 (DQA1*0501/B1*0301) and DQ9 (DQA1*0201/B1*0303), are capable of binding peptide repertoires in complementarity with DR molecules. The results reflect fundamental differences in the binding modes of these two HLA class II isotypes, in that DQ7 and DQ9 but not DR molecules appear to have the capacity to bind peptide structures without type 1-like anchor residues. Consistent with this is our observation that none of the amino acid side chains of the class II-associated invariant chain peptides (CLIP) are required for association with DQ7 and DQ9, even though many of them are essential for CLIP-DR interaction. Together, these data reveal a functional complementarity of HLA-DR and -DQ molecules in Ag presentation. PMID- 9218586 TI - Structural analysis of human antibodies to proteinase 3 from patients with Wegener granulomatosis. AB - We determined the structure of five IgM autoAbs to proteinase-3 (PR3). These Abs are highly specific for Wegener's granulomatosis (WG) and may be involved in the pathogenesis of vasculitis in WG. Five clonal lymphoblastoid cell lines secreting Abs to PR3 were derived from four patients' B cells. From 3 to 5% of supernatants from wells contained detectable anti-PR3 Abs, indicating that anti-neutrophil cytoplasmic Ab specificity represents a sizable part of the IgM B cell repertoire in patients with WG. Mu heavy chains of WG1, WG4-1, and WG4-2 clones belonged to the VH3 subgroup. WG4-1 and WG4-2 heavy chains were identical, indicating an oligoclonal expansion of autoreactive B cells in this patient. WG4-1 (and WG4-2) used the VH3-23 V(H) gene, the product of which was shown to directly bind PR3. Heavy chains of WG2 and WG3 derived from VH4-59 and VH1-2 genes, respectively. Comparison with germline sequences showed that three of the five V(H) genes from clonal lines were somatically mutated with a R:S ratio in complementarity determining regions of 3:0, 5:1, and 5:1, respectively. Three kappa light chains derived from the Vkappa4 gene, and one derived from a Vkappa1 gene. In these four Vkappa genes, there were overall R:S ratios of mutation of 8:1 and 0:7 in complementarity-determining regions and framework regions, respectively. These data suggest that the production of these autoantibodies, which are increasingly important in the diagnosis and management of WG, are influenced by an Ag-driven process. PMID- 9218587 TI - A cloned chicken lymphokine homologous to both mammalian IL-2 and IL-15. AB - IL-2, cloned in several mammalian species, plays a critical role in immune system function. Indirect evidence suggests that IL-2-like molecules also exist in lower vertebrates, although none has been cloned. In view of the commercial importance of poultry and the lack of any IL-2 sequence data in lower vertebrates, we undertook to clone a chicken IL-2-like molecule. To this end, a cDNA library was constructed from activated chicken spleen cells and screened for T cell proliferative activity. Three clones of a single gene were isolated that produced a protein stimulatory for activated T cell blasts. This protein had 24 to 25% amino acid identity and 44 to 46% similarity to both bovine IL-2 and IL-15. The homology with IL-15 was unexpected, since primate IL-15 was reported to have no homology with IL-2. However, genetic distance analysis indicated that the chicken interleukin homology is closer to the mammalian IL-2 than to the mammalian IL-15 homology. Furthermore, the chicken gene has several characteristics of mammalian IL-2s: e.g., expression only by activated T cells, mRNA with a short 5' region preceding the open reading frame, and a short leader sequence. It has one characteristic that is unique to IL-15: four conserved cysteines allowing for two intrachain disulfide bonds. PMID- 9218588 TI - Transgenic expression of a novel thymic epithelial cell antigen stimulates abberant development of thymocytes. AB - We previously reported a novel thymic stromal cell Ag, HS9, as a potent molecule participating in intrathymic T cell development. HS9 Ag is expressed on thymic stromal cells especially in the cortex but not on thymocytes. In the present study, we isolated and characterized a novel cDNA, N14, encoding HS9 Ag. Sequencing analysis of N14 cDNA has revealed it to be a novel one without any significant homology to previously reported functional molecules. COS7 cells transfected with expression vectors harboring N14 cDNA became reactive with HS9 specific mAb. Northern blot analysis and in situ hybridization revealed that several tissues that are positive for HS9 mAb expressed N14 mRNA. To examine the role of this molecule in T cell development, transgenic mice were generated. In situ hybridization and immunohistochemical study showed that the transgene was significantly overexpressed on both cortical and medullar thymic stromal cells but not on thymocytes. Flow cytometric analyses showed that the percentages of mature CD4- CD8+ or CD4+ CD8- thymocytes in transgenic mice were approximately twice and triple, respectively, those in control littermates. Moreover, substantial CD4+ CD8+ thymocytes appeared to have high levels of TCR compared with peripheral T cells. Histologic examination revealed that transgenic mice had thin cortex and relatively developed medulla. These data indicate the critical role of the N14 gene in T cell development. PMID- 9218589 TI - Evolution of proteasome subunits delta and LMP2: complementary DNA cloning and linkage analysis with MHC in lower vertebrates. AB - The class II region of the mammalian MHC harbors two proteasome subunit genes, LMP2 and LMP7. These genes are induced by IFN-gamma, and their products are incorporated into proteasomes substituting for their closest relatives, the delta and X subunits, respectively. This substitution is believed to change the proteolytic specificity of proteasomes, making it more suitable for generation of peptides to be presented by class I molecules. To elucidate the phylogenetic origin of LMP2 and the linkage of its gene with the MHC, reverse transcriptase PCR amplification of Xenopus laevis and lamprey liver mRNA was performed with primers designed to amplify both the mammalian LMP2 and delta sequences. Both LMP2 and delta were amplified from X. laevis, whereas only delta was amplified from lamprey, suggesting that delta/LMP2 gene duplication occurred after divergence of cyclostomes but before divergence of amphibians. The linkage between the LMP2 gene and the MHC was observed in a diploid Xenopus species, Xenopus tropicalis, but not in a tetraploid species, X. laevis, indicating that this linkage was established before the divergence of amphibian from higher vertebrates, but that this linkage was lost in X. laevis, probably by a gene reorganization accompanying the tetraploidization. The X. laevis LMP2 and LMP7 mRNA showed a similar tissue distribution, indicating that the genetic linkage is not required for apparently coordinated tissue-specific expression of these genes. Sequence and linkage analyses suggest that LMP2 may not play as vital a role as LMP7 in Ag presentation. PMID- 9218590 TI - A new 12-kilodalton dimer associated with pre-TCR complex and clonotype independent CD3 complex on immature thymocytes. AB - We have characterized the pre-TCR/CD3 complex and the clonotype-independent CD3 (CIC) complex expressed on the cell surface of the immature cell line KKF which we previously identified as the cell line expressing only TCR beta-chain associated with the CD3 complex on the cell surface. We now show that KKF expresses the pT alpha-beta heterodimer-CD3 complex and also the CIC complex on the cell surface by surface biotinylation. In addition, not only the CIC complex but also the pT alpha-beta complex was found to be associated with the molecular chaperon calnexin. Whereas the CD3 zeta-chain was very weakly associated with the pre-TCR/CD3 complex, a 12-kDa molecule (designated pTAC12 as pre-TCR-associated chain) was more stably associated as a dimer with the pre-TCR complex as well as the CIC complex on KKF cells. The significance of pTAC12 dimer in the CD3 complex of immature thymocytes in vivo was demonstrated by coprecipitation of pTAC12 with the CD3 complex in thymocytes from SCID mice. These results show that pTAC12 is a component of the pT alpha-beta complex as well as the CIC complex, and they suggest that pTAC12 may play a role in signaling through and/or in the expression of the pre-TCR/CD3 complex. PMID- 9218591 TI - Deviated overexpression of TCR-beta, TCR-gamma, CD4, and CD8 on thymic lymphomas induced by 1-propyl-1-nitrosourea: destruction of the allelic exclusion of TCR beta and expression of functional TCR-betagamma heterodimer on a lymphoma, cFTL53. AB - Thymic lymphomas (FTLs) induced by the chemical carcinogen 1-propyl-1-nitrosourea (PNU) in F344 rats showed deviated overexpression of TCR-beta, TCR-gamma, CD4, and CD8. Even though most FTLs were in the CD4+ CD8+ stage, all FTLs expressed TCR-beta mRNA with TCR-gamma mRNA, but without TCR-alpha mRNA or TCR-delta mRNA. One of the FTLs, cFTL53, expressed two kinds of TCR-beta mRNA and two kinds of TCR-gamma mRNA, but did not express any mRNA of TCR-alpha or TCR-delta. Both alleles of TCR-beta loci were rearranged on cFTL53. cDNA cloning and sequencing analysis showed that one TCR-gamma mRNA, Vgamma4-Jgamma1-Cgamma1, and both TCR beta mRNA, Vbeta2-Dbeta2-Jbeta2.1 and Vbeta19-Dbeta2-Jbeta2.1-Cbeta2, on cFTL53 were in the productive form, while the other TCR-gamma mRNA, Vgamma1-Jgamma4 Cgamma4L, was not. Both TCR beta-chains and a TCR gamma-chain were expressed on cFTL53, making a novel set of TCR-betagamma heterodimer. Cross-linking of TCR betagamma heterodimer on cFTL53 resulted in a calcium flux, indicating that TCR betagamma works as a signal transduction receptor. Thus, there are four strange phenomena on FTLs; CD4 and CD8 are expressed without TCR-alphabeta or TCR gammadelta, TCR-beta mRNA and TCR-gamma mRNA were expressed simultaneously without TCR-alpha and TCR-delta mRNA on FTLs, the allelic exclusion of TCR-beta was destroyed in cFTL53, and a novel set of functional TCR-betagamma heterodimer was expressed on cFTL53. PMID- 9218592 TI - Normal junctional diversification of immune receptors in p53-deficient mice. AB - One of the strategies that the immune system utilizes to generate Ab and TCR diversity is programmed imprecision of coding joint formation. This is accomplished by both nucleotide loss and random nucleotide addition (N segments) to the termini of immune receptor coding segments before resolution. Although it has been known for more than a decade that terminal deoxynucleotidyl transferase is the enzyme responsible for N segment addition, the enzymes responsible for nucleotide loss have not been identified. Recently, the p53 tumor suppressor protein was shown to have an intrinsic exonuclease activity; we reasoned that p53 as an exonuclease might be responsible for coding end processing during V(D)J recombination. Thus, we examined nucleotide loss from Ig and TCR-beta coding joints in mice lacking p53. We find no significant difference in the degree of nucleotide loss in coding joints isolated from these animals as compared with littermate controls. Thus, we conclude that p53 does not play a role in removal of nucleotides from coding termini during V(D)J recombination. Additionally, recent evidence has surfaced suggesting that p53 may play an important checkpoint role in early thymocyte differentiation. More specifically, it has been suggested that p53 is required to prevent thymocytes from maturing to the double-positive stage in the absence of a functionally rearranged TCR-beta allele. Our data suggest that TCR-beta selection is not affected in p53-deficient, V(D)J rearrangement-proficient mice. PMID- 9218593 TI - Human colorectal cancer (CRC) antigen CO17-1A/GA733 encoded by adenovirus inhibits growth of established CRC cells in mice. AB - The human colorectal carcinoma (CRC)-associated Ag CO17-1A/GA733, originally defined by mAbs CO17-1A and GA733, has been a useful target in passive immunotherapy of CRC patients with mAb and in active immunotherapy with anti idiotypic Abs mimicking the CO17-1A or GA733 epitope. Both approaches have targeted single epitopes. We investigated the capacity of full-length CO17 1A/GA733 Ag expressing multiple potentially immunogenic epitopes and encoded by recombinant adenovirus 5 (Ad5 GA733-2) to induce humoral, cellular, and/or protective immunity in mice. Ad5 GA733-2 induced Ag-specific Abs that reacted predominantly to CO17-1A- and GA733-unrelated epitopes on the Ag and lysed Ag positive CRC targets in conjunction with effector cells. Ad5 GA733-2-immune mice developed Ag-specific, proliferative lymphocytes of Th1 type and cytolytic lymphocytes. The use of Ad5 GA733-2 to immunize mice bearing established syngeneic CRC cells transfected with the human Ag induced significant and specific tumor regression. Cured mice resisted rechallenge with human CO17 1A/GA733 Ag-negative parental CRC cells, suggesting that targeting the human Ag on the murine transfectants induced protective immunity to other Ag expressed by the parental tumor. These results may explain the high potency of the recombinant vaccine. Thus, rAd5 GA733-2 may have potential as a vaccine for CRC patients. PMID- 9218594 TI - Interleukin-10 prevents dendritic cell accumulation and vaccination with granulocyte-macrophage colony-stimulating factor gene-modified tumor cells. AB - A wide variety of human tumors express IL-10 for reasons poorly understood. We have analyzed the effect of spontaneous IL-10 expression by a mouse tumor (J558L) on its immunoparalyzing effect. Because "cross-priming" of T cells by host Ag presenting cells for MHC class I-restricted tumor Ags is a major pathway for induction of tumor immunity and that is enhanced by granulocyte-macrophage (GM) CSF, we expressed this cytokine in J558L cells. GM-CSF-secreting cells were not effective when used for immunization against challenge with the parental tumor. Inhibition of IL-10 expression through an IL-10 antisense retrovirus restored the vaccine efficacy of GM-CSF-producing J558L cells, demonstrating a direct role of IL-10 in paralyzing the GM-CSF-induced antitumor immune response. Since the tumor used for challenge produced IL-10, we conclude that IL-10 interfered primarily with the initiation but not the effector phase of the immune response. Immunohistochemical analysis of the vaccine site showed a GM-CSF-induced accumulation of dendritic cells (DC) (MHC class II+ and DEC-205+) in the absence of IL-10. In the presence of IL-10, DC accumulation was completely inhibited. Together, our results demonstrate an antagonistic effect of IL-10 with respect to GM-CSF-induced DC accumulation and tumor immunity and suggest a new mechanism by which tumors escape immune recognition: namely by preventing APC from obtaining access to tumor Ags. PMID- 9218595 TI - IL-4 protects against TNF-alpha-mediated cachexia and death during acute schistosomiasis. AB - To examine the role of the Th2-type response during schistosomiasis mansoni we compared disease progression in wild type (wt), and Th2-response deficient IL-4( /-) mice. Whereas wt C57BL/6 mice tolerate infection and develop chronic disease, IL-4(-/-) C57BL/6 animals are highly susceptible, exhibiting severe acute cachexia followed by death. Data point toward morbidity in the IL-4(-/-) C57BL/6 mice being mediated by TNF-alpha, possibly through the uncontrolled production of nitric oxide in target organs such as the ileum. We propose that IL-4 prevents severe disease during schistosomiasis by regulating macrophage activation. PMID- 9218596 TI - IL-2 and IL-12 act in synergy to overcome antigen-specific T cell unresponsiveness in mycobacterial disease. AB - IL-12 secretion by APC is critical for the development of protective Th1-type responses in mycobacterial (Mycobacterium avium and Mycobacterium tuberculosis) infections in mice. We have studied the role of IL-12 and IL-2 in the generation of Mycobacterium leprae-specific T cell responses in humans. Leprosy patients were defined as low/nonresponders or high responders based on the level of T cell proliferation in M. leprae-stimulated PBMC. In high responders, M. leprae-induced proliferation was markedly suppressed by neutralizing anti-IL-12 mAb (inhibition 55 +/- 6%). Neutralization of IL-2 activity resulted in an inhibition of 77 +/- 4%. Given the importance of endogenous IL-2 and IL-12 in M. leprae-induced responses, we investigated the ability of rIL-2 and rIL-12 to reverse T cell unresponsiveness in low/nonresponder patients. Interestingly, rIL-12 and rIL-2 strongly synergized in restoring both M. leprae-specific T cell proliferation and IFN-gamma secretion almost completely to the level of responder patients. A similar synergy between rIL-2 and rIL-12 was also observed in high responders when suboptimal M. leprae concentrations were used for T cell stimulation. Our data demonstrate a crucial role for endogenous IL-12 and IL-2 in M. leprae induced T cell activation. Most importantly, we show that rIL-2 and rIL-12 act in synergy to overcome Ag-specific Th1 cell unresponsiveness. These findings may be applicable to the design of antimicrobial and antitumor vaccines. PMID- 9218597 TI - Interferons up-regulate STAT1, STAT2, and IRF family transcription factor gene expression in human peripheral blood mononuclear cells and macrophages. AB - IFN signaling is mediated by binding of IFNs to their receptors and subsequent activation of Janus tyrosine kinase (JAK)-STAT signaling pathway. Stimulation of cells with IFN-alpha leads to the assembly of IFN-stimulated gene factor 3 transcription factor complex formed by STAT1, STAT2, and p48 protein. IFN-gamma signaling is mediated by homodimeric STAT1 protein. Although these signaling molecules are expressed constitutively, there is also evidence of transcriptional regulation by IFNs. We have characterized the expression of STAT and IFN regulatory factor (IRF) family transcription factors in primary human blood mononuclear cells and macrophages in response to IFN-alpha and IFN-gamma stimulation. We show that IFN-alpha and IFN-gamma rapidly and efficiently enhanced STAT1, STAT2, p48, and IRF-1 gene expression. IFN-gamma induced IRF-1 gene expression more strongly than IFN-alpha. Stimulation experiments in the presence of protein synthesis inhibitor, cycloheximide, suggested that these genes were activated directly by IFNs. IRF-2 gene was apparently only weakly responsive to IFNs in these cells. When macrophages were pretreated with low doses of IFN-gamma and then stimulated with IFN-alpha, clearly enhanced formation of specific transcription factor complexes was detected. This suggests that higher intracellular levels of STAT1, STAT2, and p48 protein may result in enhanced signal transduction for cytokines utilizing these transcription factors. PMID- 9218598 TI - Role of adhesion molecules of the selectin-carbohydrate families in antibody dependent cell-mediated cytoxicity to schistosome targets. AB - The role of adhesion molecules in antibody-dependent cell-mediated cytotoxicity (ADCC) of macrophages toward the extracellular parasite Schistosoma mansoni was investigated by using 1) a panel of blocking mAbs directed against adhesion molecules and 2) different soluble ligands as candidate inhibitors of ADCC. The results show that the beta2-integrin Mac-1 (CD11b/CD18), L-selectin (CD62-L), and the carbohydrate determinant sialyl Lewis(x) (sLe(x); sCD15) are required for macrophage effector function toward schistosomula targets. On the other hand, the parasite counter-receptors involved in ADCC were found to share common motifs with the mammalian selectin-carbohydrate families. One family of parasite receptor(s) involved in ADCC carries the Lewis(x) (Le(x); CD15) carbohydrate structure, whereas a second family of receptor(s) appears to display selectin like properties with affinity for the sLe(x) tetrasaccharide. Immunostaining experiments confirmed that schistosomula express on their surface hostlike molecules recognized by anti-Le(x) (CD15) and by anti-human E-selectin (CD62-E) mAbs. The double receptor-ligand interaction between macrophages and parasite targets provides new insights into the biologic roles of selectins and Le(x) related structures in immunity against helminthic parasites. PMID- 9218600 TI - CD50 monoclonal antibodies inhibit neutrophil activation. AB - The constitutive high expression of CD50 (ICAM-3) on resting leukocytes, coupled with the observation that CD50 is the primary LFA-1 ligand on resting T cells, suggests that CD50 may be an important LFA-1 ligand in the initiation of the immune/inflammatory response. CD50 mAbs have been reported to increase homotypic adhesion of lymphocytes, and lymphocyte adhesion to HUVEC and extracellular matrix proteins. In this study, the effects of CD50 mAbs on neutrophil activation were examined. CD50 mAbs were found to inhibit neutrophil adhesion induced by FMLP and 12-O-tetradecanoyl-phorbol-13-acetate to resting and TNF-activated HUVEC. CD50 mAbs also inhibited neutrophil adhesion stimulated by CD66a, CD66b, CD66c, and CD66d mAbs to HUVEC. CD50 mAbs inhibited the up-regulation of CD11b/CD18 to the neutrophil surface, and the down-regulation of surface CD62L expression. The potential contribution of src family kinases to the previously described tyrosine kinase activity associated with CD50 in neutrophils was also examined. hck and lyn were found to account for much of the tyrosine kinase activity associated with CD50 in neutrophils. The data indicate that CD50 in neutrophils functions not only as a potential ligand for LFA-1, but also regulates the surface expression and activity of CD11b/CD18 and CD62L. In contrast to the effects in lymphocytes, CD50 appears to function as a negative regulator of neutrophil activation. PMID- 9218599 TI - The CDK inhibitor, p27Kip1, is required for IL-4 regulation of astrocyte proliferation. AB - IL-4 is a pleiotrophic cytokine that has been shown to affect cells of the central nervous system. We have demonstrated that IL-4 inhibits DNA synthesis and proliferation in human astroglia expressing IL-4 receptors. In this study, we sought to identify mechanisms that could account for the antimitogenic effects of IL-4. Epidermal growth factor (EGF)-stimulated human astroglia were arrested in G1 phase by IL-4, even though IL-4 stimulated levels of the G1 cyclins, D1 and E. Histone H1 kinase activity of cdk2 immunoprecipitates, however, was sharply reduced by IL-4; impairment of kinase activity was also evident in cyclin E immunoprecipitates, which contained evidence of hypophosphorylated (inactive) cdk2 product. Reduced cyclin E-associated cdk2 activity was not due to impaired cyclin-dependent kinase-activating kinase (CAK) activity, which was unaffected by IL-4. Inactive cyclin E/cdk2 complexes from IL-4 + EGF-treated cells contained, however, strikingly elevated p27Kip1 cdk inhibitor. Elevated p27 was also detectable in whole cell lysates after 24 and 48 h of IL-4 treatment; by 72 h, p27 was no longer elevated. Pretreatment with antisense but not mismatch p27 oligonucleotides attenuated the inhibitory effects of IL-4 on DNA synthesis and histone kinase activity of cyclin E/cdk2 complexes. Antisense p27 also abrogated IL-4-mediated elevation of p27 in whole cell lysates and cyclin E/cdk2 complexes. These findings demonstrate that IL-4 regulates the cell cycle machinery of astroglial cells via a p27Kip1 braking mechanism. PMID- 9218601 TI - Activation of phosphatidylinositol 3'-kinase by insulin-like growth factor-I rescues promyeloid cells from apoptosis and permits their differentiation into granulocytes. AB - Insulin-like growth factor-I (IGF-I) promotes cell division and prevents programmed cell death in hemopoietic progenitors. Human HL-60 promyeloid cells differentiate toward the granulocytic lineage when stimulated with retinoic acid (RA) in serum-containing medium. When deprived of serum, however, we found that these cells differentiate poorly in the presence of RA, as assessed by expression of the alpha subunit of the beta2 integrin heterodimer, CD11b/CD18. However, when IGF-I is added to RA-treated cells, the proportion of CD11b-positive cells increases to a level similar to that in RA-treated cells cultured in serum containing medium. Cells treated with RA alone not only differentiate poorly but also undergo apoptosis, as assessed by flow cytometry using propidium iodide and HO33342. In serum-free medium, one-third of RA-treated cells become apoptotic compared with only 5% apoptotic cells in the absence of RA. However, addition of IGF-I to RA-treated cells prevents the appearance of this apoptotic population and increases phosphatidylinositol 3'-kinase (PI 3-kinase) activity by fivefold. Wortmannin, a PI 3-kinase inhibitor, potently decreases this IGF-I-induced lipid kinase activity, blocks the ability of IGF-I to prevent apoptosis, and inhibits IGF-I-enhanced CD11b expression. These data demonstrate that IGF-I acts on RA treated progenitors to promote their differentiation along the granulocytic lineage. IGF-I acts by rescuing these cells from apoptotic cell death via a downstream pathway that is dependent upon PI 3-kinase. PMID- 9218602 TI - Recombinant human granulocyte-macrophage colony-stimulating factor (CSF), but not recombinant human granulocyte CSF, down-regulates the recombinant human stem cell factor-dependent differentiation of human fetal liver-derived mast cells. AB - The effects of recombinant human granulocyte CSF (rhG-CSF) and recombinant human granulocyte-macrophage CSF (rhGM-CSF) on the recombinant human stem cell factor (rhSCF)-dependent development of human mast cells from fetal liver progenitors were examined. Mast cells were identified by immunohistochemical staining for tryptase and by flow cytometric analysis of surface Kit expression. Only rhGM-CSF affected mast cell development. When rhGM-CSF (1, 10, or 100 ng/ml) and rhSCF (50 ng/ml) were added to cell cultures from day 0, both the percentage and absolute numbers of mast cells were diminished after 4 wk compared with cultures exposed to rhSCF alone. Half of the maximal response was achieved at a dose of rhGM-CSF between 0.1 and 1 ng/ml. The Kit+ cells developing in the presence of rhGM-CSF and rhSCF exhibited an intensity of surface Kit expression comparable to that of cells exposed to rhSCF alone. Also, if the initial exposure to rhGM-CSF was delayed for 1 to 3 wk, attenuation of mast cell development waned. These findings are consistent with uncommitted progenitor cells being diverted to nonmast cell lineages by rhGM-CSF, while cells committed to a mast cell lineage, albeit immature, appear to be resistant to the lineage directives of rhGM-CSF. Exposure of fetal liver cells to rhGM-CSF for 1 to 3 days before addition of rhSCF further diminishes the number of mast cells that develop compared with the simultaneous addition of these growth factors on day 0. Whether administration of rhGM-CSF to humans before or together with rhSCF diminishes the mast cell hyperplasia that occurs with rhSCF alone remains to be determined. PMID- 9218603 TI - IL-4 and IL-10 modulation of CD40-mediated signaling of monocyte IL-1beta synthesis and rescue from apoptosis. AB - Previous studies have demonstrated that the interaction of CD40 on monocytes with CD40 ligand, present on activated CD4+ T cells, induces monocyte inflammatory cytokine synthesis and rescues monocytes from apoptosis. These findings suggest a role for CD40 signaling of monocyte activation in the maintenance and/or exacerbation of nonseptic (e.g., autoimmune) inflammatory responses. In the present study the effects of the modulatory cytokines IL-4 and IL-10 on CD40 mediated signaling of monocyte IL-1beta synthesis and rescue from apoptosis were examined. Both IL-4 and IL-10 decreased CD40-dependent IL-1beta synthesis in a dose-dependent manner individually and synergized in this effect when used concurrently, with minimal effect on CD40 surface expression. CD40 signaling of IL-1beta synthesis was shown to be dependent on the induction of protein tyrosine kinase (PTK) activity, and both IL-4 and IL-10 diminished CD40-mediated tyrosine phosphorylation of monocyte cellular proteins. However, IL-4, but not IL-10, blocked CD40-mediated rescue from apoptosis, an event that we have demonstrated previously to be dependent on PTK activity as well. Together these results suggest that in monocytes 1) both IL-4 and IL-10 target CD40-induced PTK activity in the down-regulation of IL-1beta synthesis; and 2) IL-4 and IL-10 have divergent effects on the CD40 signaling pathway, in that these cytokines are synergistic with respect to their abilities to inhibit CD40-mediated IL-1beta synthesis and differ in their abilities to block CD40-mediated rescue from apoptosis. PMID- 9218604 TI - The role of gammadelta T lymphocytes in lipopolysaccharide-induced eosinophil accumulation into the mouse pleural cavity. AB - LPS induces an accumulation of eosinophils in the pleural cavity that requires resident macrophages and lymphocytes, but is independent of IL-5 production. In the present study we investigated the involvement of different T lymphocyte subsets on the modulation of LPS-induced eosinophil accumulation into the pleural cavity of mice. Within 4 h after LPS injection the number of neutrophils in the pleural cavity increased significantly. Mononuclear cell counts increased after 12 h, while a significant rise on eosinophil counts was observed only after 24 h. T lymphocytes counts were increased in the pleural cavity 24 and 48 h after LPS administration. This T lymphocyte accumulation was accounted for by an influx of the gammadelta+ subset, while CD4+ and CD8+ subsets did not accumulate in the pleural cavity after LPS stimulation. All those changes had resolved 96 h after LPS injection. Depletion of T lymphocytes by treatment with mAb anti-Thy 1.0 inhibited the eosinophil accumulation triggered by LPS. Aiming to clarify which T lymphocyte subset would be involved in the LPS-induced eosinophil accumulation, we depleted mice of various T lymphocyte subpopulations using specific Abs. Depletion of either CD4+ or CD8+ subsets failed to inhibit LPS-induced eosinophil migration. In contrast, when mice were treated with anti-gammadelta+ T lymphocyte mAb, a significant reduction of LPS-induced eosinophil accumulation was observed. Similarly, the administration of LPS in BALB/c-nu/nu mice induced the expected significant influx of eosinophils into the pleural cavity. Our results indicate that the gammadelta+ T lymphocytes are centrally involved in LPS-induced eosinophil accumulation in mice. PMID- 9218605 TI - TNF-alpha-mediated expression of the receptor for anaphylatoxin C5a on neurons in experimental Listeria meningoencephalitis. AB - The anaphylatoxin C5a has been implicated in the pathogenesis of bacterial meningitis as a potent mediator of inflammation in the subarachnoid space. We investigated the expression of the receptor for C5a (C5aR) in brains of mice with experimental Listeria monocytogenes (LM) meningoencephalitis. In the course of the disease, infiltrating cells in the meninges and the ventricles were found to express C5aR mRNA and protein. In the brain parenchyma, very low constitutive C5aR expression was seen on pyramidal neurons and Purkinje cells. However, in LM infected mice, a dramatic increase in C5aR expression occurred on neurons starting 6 h after infection and was maximal between 24 and 36 h. TNF-alpha was identified as an essential mediator of neuronal C5aR expression, since mice lacking the genes for TNF and lymphotoxin-alpha (TNF/lymphotoxin-alpha -/- mice) showed significantly attenuated C5aR expression after LM infection. Furthermore, i.p. injection of recombinant TNF-alpha induced enhanced C5aR expression in the brains of TNF/lymphotoxin-alpha -/- mice and in normal animals even in the absence of a bacterial infection. We also assessed the levels of anaphylatoxin C5a in the cerebrospinal fluid of patients with infectious meningitis. C5a was detected in all patients with bacterial meningitis (n = 9), in 6 of 18 patients with aseptic meningitis, and in 1 of 66 control patients. The finding of TNF alpha-mediated C5aR expression on neurons in experimental Listeria meningitis and the detection of the ligand, C5a, in the cerebrospinal fluid of human patients with infectious meningitis present new directions in the investigation of the pathophysiologic sequelae leading to secondary brain damage. PMID- 9218606 TI - Chemokine expression in experimental tubulointerstitial nephritis. AB - Chemokines may be important in the pathogenesis of leukocyte infiltration in tubulointerstitial nephritis associated with glomerular disease. We studied the renal cortical expression of the C-C (macrophage inflammatory protein-1alpha (MIP 1alpha)), monocyte chemotactic protein-1 (MCP-1), and RANTES) and C-X-C (interferon-inducible protein-10 (IP-10), MIP-2, and cytokine-induced neutrophil chemoattractant (CINC)) chemokines 4, 6, 8, 10, 14, and 21 days after the induction of puromycin aminonucleoside (PAN) nephrosis. There was a 7- to 10-fold increase in the steady state mRNA expression of IP-10 and MCP-1 in the renal cortex of rats 6 to 8 days after the administration of PAN that declines thereafter reaching control values by day 21. The site of IP-10 and MCP-1 mRNA production was localized to intrinsic tubulointerstitial cells and not to infiltrating monocytes or macrophages. By comparison, there was a low basal expression of RANTES mRNA in the renal cortex of nephrotic rats that did not differ from those of control rats. In contrast, CINC, MIP-2, and MIP-1alpha mRNAs were not detected. Translation of MCP-1 mRNA into protein was confirmed with an ELISA. These changes in chemokine gene expression were associated with a tubulointerstitial T lymphocyte and macrophage infiltration beginning on day 6 that peaked on day 10. Administration of a neutralizing Ab to rat MCP-1 (n = 5) beginning on day 4 resulted in a 45% decline in tubulointerstitial macrophage infiltration from 8.4 +/- 1.3% to 4.6 +/- 0.4% (p < 0.001) on day 6. These data provide evidence that MCP-1, and possibly IP-10, are important in the pathogenesis of monocyte/macrophage infiltration in the tubulointerstitial nephritis associated with PAN nephrosis. PMID- 9218607 TI - STAT5 involvement in the differentiation response of primary chicken myeloid progenitor cells to chicken myelomonocytic growth factor. AB - We have investigated the activation of the STAT5 protein during the differentiation of myelomonocytic cells. In the human U937 promonocytic cell line, STAT5 activation occurred in response to several inducers of monocytic differentiation (phorbol ester, 1alpha,25-dihydroxyvitamin D3, and retinoic acid). In the promyelocytic HL60 cell line, STAT5 was activated in the course of either phorbol ester-induced monocytic differentiation or DMSO-induced granulocytic differentiation. To test for involvement of STAT5 in the differentiation of primary nonimmortalized cells, chicken myeloid progenitor cells transfected with the ts21-E26 avian retrovirus were studied. At 37 degrees C the temperature-sensitive oncoprotein of the ts21-E26 virus (p135(gag-myb-ets)) installs a differentiation block that is released by a shift to the nonpermissive temperature (42 degrees C). Both proliferation and differentiation of ts21-E26 transfected myeloblasts require the continuous presence of chicken myelomonocytic growth factor (cMGF). Addition of cMGF to growth factor-starved cells rapidly caused STAT5 tyrosine phosphorylation, DNA binding, and transcriptional activity at both permissive and nonpermissive temperatures. Moreover, the temperature shift-induced onset of myelomonocytic differentiation strictly correlated with the appearance of activated STAT5 in ts21-E26-infected myeloblasts, but not in cells infected with wtE26 retrovirus. These data position STAT5 in a nuclear signaling cascade induced by the cMGF receptor and suggest a contribution of STAT5 to the process of myelomonocytic differentiation or to the functional changes that accompany the maturation of myeloid progenitor cells to a terminally differentiated stage. PMID- 9218608 TI - Stimulation of Fc gamma receptors in rat peritoneal macrophages induces the expression of nitric oxide synthase and chemokines by mechanisms showing different sensitivities to antioxidants and nitric oxide donors. AB - The induction of nitric oxide (NO) production and the expression of cytokine induced neutrophil chemoattractant (CINC-1) were studied in rat peritoneal adherent cells stimulated with insoluble immune complexes containing rabbit IgG Ab and OVA as the cognate Ag (IC). Incubation with IC at concentrations as low as 10 microg/ml induced NO production and the expression of inducible NO synthase (iNOS) protein. This was accompanied by the expression of CINC-1 mRNA and the activation of nuclear factor-kappaB (NF-kappaB). However, the expression of iNOS and CINC-1 mRNA induced by IC showed a different temporal pattern and a different sensitivity to both the antioxidant agent pyrrolidine dithiocarbamate (PDTC) and modulation by NO itself. Whereas iNOS mRNA and protein expression were blunted by PDTC and NO-generating compounds, CINC-1 mRNA expression was either enhanced or not affected by PDTC and NO donors. The time course of NF-kappaB activation was parallel to that of iNOS induction and was influenced in the same sense as iNOS induction by antioxidants, NO donors, the protease inhibitor N-tosyl phenylalanine chloromethyl ketone, and inhibitors of protein tyrosine phosphorylation reactions. These data indicate the existence in rat macrophages of a signaling mechanism triggered by Fc gammaR occupancy that leads to nuclear signaling, is initiated by protein tyrosine phosphorylation reactions, and shows specific sensitivities to antioxidants and NO. Whereas trans-activation of the iNOS gene can be fully explained by the stimulation of NF-kappaB, induction of CINC-1 mRNA expression seems influenced by additional regulatory elements. PMID- 9218609 TI - Suppressive effects of recombinant human monokine induced by IFN-gamma (rHuMig) chemokine on the number of committed and primitive hemopoietic progenitors in liquid cultures of CD34+ human bone marrow cells. AB - Studies in this report investigated potential hemopoietic suppressive effects of human monokine induced by IFN-gamma (HuMig), a CXC chemokine that is chemotactic for activated lymphocytes. rHuMig was purified from Trichoplusia ni cells after infection with a recombinant baculovirus. The recombinant protein was added to liquid cultures of CD34+ human marrow cells stimulated with IL-3 alone or with both IL-3 and either insulin-like growth factor II (IGF-II) or stem cell growth factor (SCF). The number of committed progenitors, colony-forming units for granulocytes and macrophages (CFU-GM), and primitive progenitors, long term culture-initiating cells (LTC-IC) derived from liquid cultures of CD34+ cells, was determined. rHuMig abrogated the IGF-II-dependent enhancement of CFU-GM and long term culture-initiating cell numbers. Additional studies demonstrated that in liquid cultures of CD34+ cells both rHuMig and IFN-inducible protein-10, another CXC chemokine that is related to HuMig, inhibited the production or expansion of CFU-GM. For a subset of marrows, rHuMig also abrogated SCF enhancement of CFU-GM numbers in cultures of CD34+ cells stimulated with both IL 3 and SCF. These studies are the first to demonstrate that rHuMig can act as a negative regulator of in vitro hemopoiesis, that both rHuMig and IP-10 can suppress an increase in the number of committed progenitors from CD34+ cells, and that chemokines can abrogate hemopoietic stimulatory effects of IGF-II. PMID- 9218610 TI - Functional expression of the CXC-chemokine receptor-4/fusin on mouse microglial cells and astrocytes. AB - The mRNA for the seven-transmembrane-spanning G protein-coupled receptor fusin/CXCR-4 is expressed in primary mouse astrocyte cultures and the transformed mouse microglial cell line, N9. Cell surface expression of fusin in these cells was confirmed by staining with a polyclonal anti-fusin Ab. The functional capacity of this chemokine receptor was examined by evaluating the calcium responses following stimulation of glial cells with the CXC-chemokine, stromal derived cell factor-1alpha (SDF-1alpha). Both astrocytes and microglial cells mobilized calcium following stimulation with chemically synthesized SDF-1alpha. SDF-1alpha- and carbachol-mediated calcium responses of astrocytes were partially inhibited by treatment with pertussis toxin (PTx), suggesting receptor coupling to a combination of G alpha(i) and other G proteins. In contrast, the calcium responses of microglial cells to SDF-1alpha were completely PTx sensitive, while responses to carbachol stimulation were PTx resistant. The ability of SDF-1alpha to induce glial cell migration was also examined. Synthetic SDF-1alpha was a potent chemoattractant for mouse microglial cells at ligand concentrations of 10 to 500 ng/ml; peak responses were noted at 100 ng/ml. In contrast, astrocytes did not migrate toward a gradient of SDF-1alpha. The failure of SDF-1alpha to induce astrocyte migration was specific, as another chemokine, macrophage inflammatory protein-1alpha, triggered astrocyte chemotaxis. PMID- 9218611 TI - Cytokine-induced neutrophil-mediated injury of human endothelial cells. AB - To understand the pathogenesis of vasculitides, we analyzed how cytokine stimulation of HUVEC in vitro activates the cytotoxic capacity of polymorphonuclear (PMN) granulocytes. IL-1beta, IFN-gamma, or TNF-alpha caused highly significant dose and time-dependent HUVEC injury. TNF-alpha-treated HUVEC activated the PMN by means of phospholipase C-related event, since coincubations conferred PMN to react with a rise of cytosolic calcium concentrations, [Ca2+]i. Ab blockade of ICAM-1 on HUVEC inhibited 50 to 70% of the injury induced by these cytokines, whereas a mAb to E-selectin reduced 45 to 65% of IL-1beta- and TNF alpha-, but not IFN-gamma-induced cytotoxicity. The role of nitric oxide (NO) was of significance since injury induced by each cytokine was reduced by 60 to 87% by specific NO-synthase inhibitors, as well as by scavenging extracellular NO by oxyhemoglobin. In contrast, injury induced by TNF-alpha was inhibited by neither superoxide dismutase or catalase, alpha1-antitrypsin, alpha2-macroglobulin, nor the platelet-activating factor receptor antagonist WEB-2086. Moreover, PMN from a patient with chronic granulomatous disease were fully capable of mediating cytotoxicity. The possibility that IL-8, produced by HUVEC in response to TNF alpha, mediated activation of PMN was not corroborated since addition of an IL-8 blocking mAb did not modify HUVEC injury. Nonetheless, the IL-8 mAb (but not WEB 2086) blocked the rise of [Ca2+]i. Thus, in this in vitro model of vasculitis, the effect of IL-1beta, IFN-gamma, and TNF-alpha as promotors of cytokine mediated neutrophil-dependent injury to HUVEC is a process dependent on expression of adhesion molecules and probably associated with NO produced in the system. PMID- 9218612 TI - CD44 regulates phagocytosis of apoptotic neutrophil granulocytes, but not apoptotic lymphocytes, by human macrophages. AB - Phagocytosis of apoptotic neutrophil granulocytes by macrophages at inflammatory sites is an important determinant of the process by which inflammation resolves. We demonstrate that phagocytosis of apoptotic neutrophils, but not apoptotic lymphocytes, by human monocyte-derived macrophages is augmented rapidly following ligation of CD44 by bivalent Abs in vitro. Previously defined inhibitors of apoptotic cell recognition did not affect CD44-augmented phagocytosis of apoptotic neutrophils, suggesting that unique molecular recognition pathways are involved. These observations, together with the lack of effect of CD44 Abs upon macrophage phagocytosis of zymosan or Ig-opsonized erythrocytes, imply that CD44 may regulate the differential clearance of apoptotic leukocytes during evolution of inflammatory responses. This represents a novel role for CD44 in inflammation and tissue repair. PMID- 9218613 TI - Roles of CD9 molecules in survival and activation of human eosinophils. AB - CD9 is a cell surface glycoprotein belonging to the transmembrane 4 superfamily. In this report, we demonstrate that cross-linking CD9 with different forms of mAb activates distinct functions of human eosinophils. Anti-CD9 mAb (clone ALB6) immobilized onto tissue culture plates induced eosinophil degranulation. This effect of anti-CD9 mAb (clone ALB6) was unique because other immobilized clones of anti-CD9 mAb (clones FMC56 and ML13) or anti-HLA Class I mAb failed to induce degranulation. In addition, neutrophil degranulation was not provoked by the immobilized clone ALB6, consistent with a lack of expression of CD9 on neutrophils. Eosinophil degranulation induced by anti-CD9 mAb (clone ALB6) was abolished by pretreatment of eosinophils with anti-CD18 mAb, suggesting that beta2 integrins are involved in the reaction. In contrast to the results with immobilized clones, all clones of anti-CD9 mAb in soluble form enhanced eosinophil survival. This enhanced survival was inhibited by anti-granulocyte macrophage-CSF (GM-CSF) mAb, suggesting that autocrine production of GM-CSF by eosinophils mediated the enhanced survival. In fact, by ELISA and RT-PCR, GM-CSF protein and mRNA were detected in supernatants and cell lysates of eosinophils stimulated by soluble anti-CD9 mAb, but not by immobilized mAb. These results indicate that CD9 serves as a molecule that delivers stimulation signals on eosinophils. The eosinophil's cellular responses may differ and be dependent on the manner of receptor ligation. PMID- 9218614 TI - CD7-mediated regulation of integrin adhesiveness on human T cells involves tyrosine phosphorylation-dependent activation of phosphatidylinositol 3-kinase. AB - The functional activity of integrin receptors on T cells is dynamically regulated so that T cells can alternate rapidly between adhesive and nonadhesive states. The CD7 Ag is one of several molecules on T cells that can transduce intracellular signals that rapidly up-regulate integrin-mediated adhesion. We demonstrate in this report that the signaling pathway that CD7 utilizes to regulate integrin activity involves the lipid kinase phosphatidylinositol 3 kinase (PI 3-K). CD7 stimulation of both Jurkat T cells and resting human peripheral blood CD4+ T cells results in rapid association and activation of PI 3 K with CD7. Phosphopeptide competition assays demonstrate that the association of CD7 with PI 3-K is dependent on tyrosine phosphorylation of the SH2 binding motif Tyr-Glu-Asp-Met (YEDM) in the CD7 cytoplasmic domain. A role for PI 3-K in the regulation of integrin function by CD7 is demonstrated by: 1) the ability of two structurally distinct PI 3-K inhibitors, wortmannin and LY294002, to inhibit CD7 mediated increases in beta1 integrin function of human T cells; and 2) inhibition of CD7-mediated activation of beta3 integrin function in human T cells by expression of a dominant negative form of the p85 subunit of PI 3-K. These results demonstrate that the CD7 Ag on human T cells is coupled to PI 3-K and that this association is relevant to CD7-mediated signaling events, specifically CD7-induced increases in integrin adhesiveness. Furthermore, these studies provide important new evidence implicating PI 3-K in the regulation of integrin adhesiveness by multiple cell surface signaling receptors. PMID- 9218615 TI - Binding of human peripheral blood polymorphonuclear leukocytes to E-selectin (CD62E) does not promote their activation. AB - E-selectin (CD62E) is a cytokine-inducible endothelial cell adhesion molecule that tethers polymorphonuclear leukocytes (PMNs) and supports PMN rolling under conditions of flow. We examined whether interaction of PMNs with E-selectin also leads to activation of CD11b/CD18 (Mac-1, alphaMbeta2), an event that can promote firm adhesion. PMNs were added to monolayers of IL-1beta-activated HUVECs and Chinese hamster ovary (CHO) cells transfected with E-selectin cDNA. PMN activation was assessed by 1) increased CD11b/CD18 surface expression, 2) appearance of activation epitope on CD11b/CD18 (CD11b*) detected by mAb CBRM1/5, and 3) decreased L-selectin (CD62L) expression, as determined by flow cytometry. Both adherent and nonadherent supernatant PMNs became activated on IL-1beta pretreated HUVECs. This activation was not affected by CD62E-blocking mAb P6E2. The activation state of PMNs adhered to CHO cells transfected with E-selectin cDNA was not increased over background and was similar to that of PMNs exposed to parent CHO cells. The findings were confirmed using confocal microscopy, which allowed staining of the cells for CD11b* in situ. In concert, the results suggest that PMN binding to E-selectin does not elicit inter-receptor signaling that could result in strengthening of PMN adhesion to endothelium. PMID- 9218616 TI - Ligation of L-selectin through conserved regions within the lectin domain activates signal transduction pathways and integrin function in human, mouse, and rat leukocytes. AB - L-selectin is a cell surface adhesion receptor that mediates leukocyte rolling along vascular endothelium at sites of inflammation and lymphocyte attachment to endothelial cells within peripheral lymphoid tissues. Since ligation of L selectin through its ligand recognition region may mimic physiologic ligand binding, a new panel of mAbs that engaged a conserved ligand-binding region within the lectin domains of human, mouse, and rat L-selectin were generated using L-selectin-deficient mice. Indeed, appropriate ligation of L-selectin generated transmembrane signals that resulted in immediate intercellular adhesion following cell-cell contact of lymphocytes, neutrophils, and L-selectin cDNA transfected cells. Ab binding to only some epitopes within the lectin domain of L selectin induced adhesion, while mAb binding to numerous other epitopes or other domains of L-selectin had no effect. PPME (yeast polyphosphomonoester core polysaccharide), a complex carbohydrate that mimics the natural L-selectin ligand, also induced potent intercellular adhesion. The induction of intercellular adhesion required cellular energy, an intact cytoskeleton, and cytoplasmic kinase activity as well as the membrane proximal and cytoplasmic domains of L-selectin. Therefore, ligation of L-selectin through specific ligand binding epitopes identified by the mAbs generated in this study can generate transmembrane signals. Signaling through L-selectin may enhance leukocyte endothelial cell interactions by serving as an activation/priming step during rolling, thereby promoting subsequent firm cell-cell adhesion in the presence of inflammatory mediators. PMID- 9218617 TI - Effect of an ectokinase inhibitor, K252b, on degranulation and Ca2+ signals of RBL-2H3 cells and human basophils. AB - We examined the effects of K252b, an ectoprotein kinase inhibitor of microbial origin, on the activation process of RBL-2H3 cells by cross-linking of IgE receptors by the endoplasmic reticulum Ca2+-ATPase inhibitor 2,5-di(tert-butyl) 1,4-hydroquinone or by the Ca2+ ionophore A23187. Analysis of phosphorylation of ectoproteins following IgE receptor cross-linking revealed that K252b mainly inhibited the phosphorylation of a 130-kDa protein. The inhibitor simultaneously inhibited degranulation and the sustained increase in the cytosolic calcium ion concentration even after addition of Ag. In contrast, K252b did not inhibit the increase in degranulation and cytosolic calcium ion concentration caused by stimulation with 2,5-di(tert-butyl)-1,4-hydroquinone and A23187. Permeation of K252b into RBL-2H3 cells, assessed by fluorescence intensity, was very low. K252b also inhibited degranulation caused by IgE receptor cross-linking in human basophils, but did not inhibit the degranulation caused by A23187. Thus, our findings suggest that the effects of K252b may be mediated by outer surface-bound or -anchored K252b-sensitive molecules on RBL-2H3 cells and human basophils, and that the phosphorylation of ectoprotein may involve a transmembrane influx of Ca2+ by IgE receptor cross-linking. PMID- 9218618 TI - Endotoxin shock in antibody-deficient mice: unraveling the role of natural antibody and complement in the clearance of lipopolysaccharide. AB - While the significance of natural Ab is not entirely clear, one proposed role is clearance of bacterial Ags. To determine whether natural Ab was involved in clearance of endotoxin, we have examined novel strains of mice with either a total or selective deficiency in Ig. Recombinase-activating gene-2 (RAG-2(-/-)) deficient mice, which have no serum Ig due to arrested development of B cells at the pro-B stage, demonstrate increased sensitivity to endotoxin that correlates with an impaired clearance. When RAG-2(-/-) mice are reconstituted with pooled sera from normal mice, both survival and clearance of circulating endotoxin are enhanced. To further define the nature of the protective Ab, Bruton's tyrosine kinase (Btk)-deficient mice were characterized in the high dose LPS model. Like RAG-2(-/-) mice, they are highly sensitive to endotoxin and have an impaired clearance of LPS. Reconstitution of Btk(-/-) mice, which have reduced levels of IgG3 and IgM, with purified normal mouse IgM dramatically enhances their ability to clear endotoxin compared with mock (saline)-reconstituted littermates. The cellular source of natural anti-LPS IgM was identified as the peritoneal-residing B-1 cell by enzyme-linked immunospot (ELISPOT) assay. Taken together, these studies demonstrate the important role of natural Ab and complement in the clearance of pathogenic substances from the circulation. PMID- 9218619 TI - Increased susceptibility to endotoxin shock in complement C3- and C4-deficient mice is corrected by C1 inhibitor replacement. AB - Endotoxin shock is a life-threatening syndrome associated with a Gram-negative infection and mediated by a systemic inflammatory response. As a major effector of inflammation, the complement system has been implicated in both the pathogenesis and the protection from endotoxin shock. To clarify the role of complement in endotoxin shock, we have used mice totally deficient in either complement component C3 or C4. We found that both the C3- and C4-deficient mice were significantly more sensitive to endotoxin than wild-type controls. The endotoxin-challenged complement-deficient mice failed to clear endotoxin efficiently from the circulation and this led to excess consumption of C1 inhibitor protein (C1 INH), a major regulator of both complement and the contact system of blood coagulation. Replacement of C1 INH rescued the endotoxin challenged complement-deficient mice from shock and death. These findings suggest a novel therapy for treatment of endotoxemia with C1 INH protein. PMID- 9218620 TI - Role of the P2 residue of complement 1 inhibitor (Ala443) in determination of target protease specificity: inhibition of complement and contact system proteases. AB - A dysfunctional C1 inhibitor (C1 INH) from a family in whom the propositus presented with systemic lupus erythematosus but without angioedema previously was shown to have diminished inhibitory activity toward isolated C1r and C1s, and intact C1. The mutation was identified as replacement of Ala443 (P2) with Val. This study further analyzed the reactivity of this mutant and characterized two mutants with Ser or Asp at this position. Ser at P2 does not interfere with binding of target proteases. However, the mutant with Asp at this position is unable to bind C1r and beta factor XIIa, and also has a decreased rate of reaction with C1s and kallikrein. Therefore, alteration of polarity alone had no effect on binding, while a bulky and/or charged side chain was not tolerated. Although defective in inhibition of C1r and C1s, the P2 A-->V mutant had acquired the ability to complex with trypsin. It also completely retained the ability to complex with kallikrein and factor XIIa. None of the 10 individuals expressing this mutant protein has ever had angioedema. This observation, combined with normal inhibition of contact system proteases and defective inhibition of complement proteases, suggests that angioedema is caused by bradykinin generated from contact system activation. PMID- 9218621 TI - Myeloid differentiation treatment to diminish the presence of immune-suppressive CD34+ cells within human head and neck squamous cell carcinomas. AB - Within human head and neck squamous cell carcinomas (HNSCC) that produce granulocyte-macrophage CSF are CD34+ cells that exhibit natural suppressive (NS) activity. The present study aimed to identify how these NS cells mediate suppression and how to diminish their presence. CD34+ cells that were immunomagnetically isolated from fresh surgical HNSCC specimens produced a soluble product that blocked normal T cell stimulation through the TCR/CD3 complex. This inhibitory activity could be neutralized with Abs to TGF-beta1. Since prior studies showed that the CD34+ NS cells within HNSCC cancers are myelomonocytic progenitor cells, the feasibility of using cytokines that can induce myeloid cell differentiation to diminish the presence of CD34+ NS cells was tested. Adding low doses of 100 U/ml IFN-gamma plus 10 U/ml TNF-alpha to bulk cultures of dissociated HNSCC cancers diminished the frequency of CD34+ cells. Studies with CD34+ cells that were isolated from the HNSCC cancers showed that this cytokine treatment induced differentiation of the CD34+ cells predominantly into monocytic cells. The consequence of treating CD34+ NS cells with the myeloid differentiation treatment was the loss of suppressive activity, a decline in TGF beta production, and the production of TNF-alpha by the resulting monocytic cells. In HNSCC bulk cultures containing high levels of CD34+ NS activity, IFN gamma/TNF-alpha not only reduced CD34+ cell levels, but also increased the capacity of the intratumoral T cells to express the p55 IL-2R. These studies show that IFN-gamma/TNF-alpha can induce differentiation of TGF-beta-secreting CD34+ NS cells into nonsuppressive monocytic cells that secrete TNF-alpha. PMID- 9218622 TI - Involvement of laminin and its receptor in abrogation of heart graft rejection by autoreactive T cells from Trypanosoma cruzi-infected mice. AB - Extracellular matrix ligands and receptors have been identified as determining in vivo lymphocyte positioning and activation, including effector functions in alloreactive responses. Herein we evaluated the involvement of laminin and its receptor, the very late antigen 6 (VLA-6) integrin, in CD4+ T cell-dependent autoreactivity, using a transplantation model for the autoimmune myocarditis occurring in mice chronically infected with Trypanosoma cruzi. Previous work showed that syngeneic mouse hearts grafted in the ears of chronic chagasic recipients were rejected through a CD4+ T cell-dependent mechanism. Rejection also occurred when cells from chagasic animals were transferred adjacent to hearts transplanted into naive recipients. Here, we observed the formation of a thick laminin network during rejection, with donor-derived CD4+ T cells concentrated in the laminin-rich areas. Most importantly, anti-laminin as well as anti-laminin receptor Ab inhibited the rejection of syngeneic hearts by T cells from chagasic animals. Our results suggest that interaction of the VLA-6 molecule with laminin is involved in triggering the antimyocardial autoreactive process by driving the influx of CD4+ T cells to the heart. They also support the concept that an Ag-specific T cell response, even an autoreactive one, can be modulated by in vivo interactions involving extracellular matrix ligands and receptors. In this regard, our study represents, to our knowledge, the first in vivo evidence for laminin-mediated T cell echotaxis, with simultaneous experimental demonstration of ligand and receptor involvement. Lastly, our findings indicate that treatment with anti-VLA-6 Abs can be effective in suppressing autoimmune disease activity. PMID- 9218623 TI - Genetic susceptibility to experimental autoimmune uveitis involves more than a predisposition to generate a T helper-1-like or a T helper-2-like response. AB - This study examines whether genetic susceptibility vs resistance to experimental autoimmune uveoretinitis (EAU) in mice is associated with dominant type 1 vs type 2 cytokine response profiles. Mice from six strains were immunized with the uveitogenic retinal Ag IRBP. EAU was evaluated by histopathology. As judged by disease scores, three of the strains were susceptible, one was minimally susceptible, and two were resistant. Ag-specific type 1 vs type 2 cytokine responses (protein and/or mRNA) in draining lymph node cells, and IgG2a vs IgG1 Ab isotypes to IRBP, were measured as indicators of Th1-like vs Th2-like responses, respectively. The three susceptible strains (B10.A, C57BL/10, and BALB/k) showed a dominant Th1-like response profile characterized by high IFN gamma and IL-12p40 (but not IL-4) responses, and a predominance of IgG2a Abs. The minimally susceptible strain (A/J) had an IFN-gamma response detectable only at the mRNA level, but produced predominantly IgG2a Abs. One of the two resistant strains (BALB/c) showed a characteristic Th2-like response with dominant Ag specific IL-4 and IL-10 responses but no IFN-gamma, and predominantly IgG1 Abs. However, the other resistant strain (AKR) did not show a Th2-dominated response pattern, in that it had low, or no, IL-4 and IL-10 responses, and made predominantly IgG2a Abs to IRBP. These results suggest that whereas a Th1 response is required for susceptibility, resistance is not dependent on a Th2 response pattern. We suggest that regulatory influences other than skewing the response toward the Th2 pathway may be equally effective at conferring genetic resistance to EAU. PMID- 9218624 TI - CD30 induction and cytokine profiles in hepatitis C virus core-specific peripheral blood T lymphocytes. AB - Since an efficient control of virus infections may depend on the appropriate lymphokine profile, we studied cytokine responses and CD30 induction, a recently proposed surrogate marker of type 2 cells, in 10 healthy anti-hepatitis C virus (HCV)-seropositive blood donors without viremia (group A) and in 15 patients with hepatitis C (group B). Intracytoplasmic IFN-gamma, IL-2, IL-4, and IL-10 were determined by triple-color flow cytometry in the CD3+ and CD3+/CD30+ lymphocyte subsets after stimulation of PBMC with rHCV core protein and five core-derived peptides corresponding to the four immunodominant Th epitopes C.T1 to C.T4. In group A, more type 1 cytokines were induced by the rHCV core protein and all immunodominant core peptides (p < 0.05), whereas IL-10-producing T cells were found more frequently in group B. Induction of CD30+ T cells was found almost exclusively in group B (p < 0.01). The difference in cytokine responses was due to the CD3+/CD30- T cell subset and not the CD3+/CD30+ subset, which predominantly produced both IL-10 and IFN-gamma, but only small amounts of IL-2 and IL-4. We conclude that immunodominant HCV core peptides induce preferentially type 1 cytokines in healthy anti-HCV-positive blood donors and CD30 expression in patients with chronic hepatitis C. However, in both groups, CD30+ T lymphocytes produce an intermediate Th0-like cytokine profile. Thus, chronicity in HCV infection may reflect a lack of type 1 cytokine production. PMID- 9218625 TI - Molecular bases of C7 deficiency: three different defects. AB - The molecular basis of C7 deficiency has been investigated in two Irish families and a number of Israeli families of Moroccan Sephardic Jewish origin. Exon PCR and sequencing revealed a heterozygous point mutation at the 3' splice acceptor site of intron 1 in one Irish family. In the other Irish family, exons 7 and 8 failed to amplify and they were shown to be deleted. Marker haplotype studies of the C6 and C7 gene region and Southern blots show that the Irish family with the splice defect also segregate for the deletion, which is not easily detected in heterozygotes. The Israeli C7-deficient cases all share a C7 haplotype and are homozygous for a mis-sense mutation in exon 9. However, one individual is heterozygous for markers at adjacent C6 loci, showing that there has been an intergenic recombination and suggesting that the deficiency mutation is of appreciable antiquity. PMID- 9218626 TI - B cell genotype determines the fine specificity of autoantibody in lpr mice. AB - Anti-Sm Abs are specific markers of human systemic lupus erythematosus (SLE) and of murine models of this disease. In humans, anti-Sm Abs are mostly IgG1, and in MRL/lpr mice, IgG2a; both are T-dependent isotypes. Other lpr strains, such as B6/lpr, do not produce anti-Sm Ab spontaneously. The present study was aimed at identifying the cellular expression of background genes responsible for generation of the anti-Sm Ab response in MRL/lpr mice. We used double chimeric mice made by transferring MRL/lpr and B6/lpr bone marrows into irradiated allotype heterozygous F1 mice. Five mo after reconstitution, FACS analysis of lymph node (LN) and spleen cells revealed that both MRL/lpr and B6/lpr cells coexisted in roughly equal numbers. Ab produced by each donor could be distinguished by allotype-specific assays. IgG2a anti-Sm was made only by MRL derived B cells despite the presence of T cells that might potentially provide help to the B6/lpr B cells. The frequency of anti-Sm Ab-producing individuals was similar to that of unmanipulated MRL/lpr mice (about 25%). IgG2a anti-chromatin and total IgG2a was mostly dominated by the MRL-derived B cells. B6-derived B cells produced more rheumatoid factor (RF) against their own IgG2b(b), while RF against IgG2a was dominated by MRL-derived B cells. This suggests that the control of the production of particular autoantibody specificities, such as anti Sm, is determined by the expression of MRL or B6 background genes in B cells. PMID- 9218627 TI - Induction of antigen-specific tolerance for the treatment of ongoing, relapsing autoimmune encephalomyelitis: a comparison between oral and peripheral tolerance. AB - Experimental autoimmune encephalomyelitis (EAE) is a T cell-mediated autoimmune demyelinating disease of the central nervous system that serves as an animal model for multiple sclerosis. Various forms of Ag-specific tolerance have been used prophylactically to prevent development of acute EAE. Here we compare the induction of Ag-specific tolerance using two regimens, proteolipid protein 139 151 (PLP139-151) peptide-coupled splenocytes and oral administration of PLP139 151, for efficacy in the reduction of established, chronic clinical EAE. PLP139 151-coupled splenocytes and not oral administration of PLP139-151 was able to down-regulate established EAE, including subsequent relapses. PLP139-151 peptide coupled splenocytes were effective at reducing Ag-specific T cell proliferation and IL-2 and IFN-gamma production, while concomitantly increasing IL-4 production. Oral administration of PLP139-151 did not reduce IL-2 or IFN-gamma production and appeared to increase Ag-specific T cell proliferation. Neither multiple high nor low doses of PLP139-151 were effective at decreasing ongoing clinical EAE or PLP139-151-specific IL-2 and IFN-gamma production. These results suggest that PLP139-151 peptide-induced tolerance is an efficacious treatment for ongoing, R-EAE when the peptide is coupled to chemically fixed splenocytes and not when given orally. PMID- 9218628 TI - Interleukin-6 is not altered in cerebrospinal fluid of first-degree relatives and patients with Alzheimer's disease. AB - We investigated interleukin-6 (IL-6) levels in cerebrospinal fluid (CSF) of 25 patients with clinically diagnosed sporadic Alzheimer's disease (AD) and 19 healthy control subjects (HC). For comparison 19 clinically healthy subjects with at least one first-degree relative with clinical or autopsy confirmed AD (CF/AD) were examined. CSF levels of IL-6 did not show statistically significant differences between AD patients, CF/AD and HC subjects. There was no correlation between age, gender, age of onset, degree of cognitive impairment, blood-brain barrier dysfunction and IL-6 values. We could not demonstrate altered CSF concentrations of IL-6 that may indicate an inflammatory response or capability to support neuronal survival in the central nervous system (CNS) of first-degree relatives and patients with AD. We suggest that combined measurement of all parameters of the IL-6-receptor complex could yield more insight in a probably altered IL-6 function. PMID- 9218629 TI - Trimethyltin induces gelatinase B and urokinase in rat brain. AB - The neurotoxicant trimethyltin (TMT) increases mRNA levels for cytokines, tumor necrosis factor-alpha, interleukin-1alpha, and interleukin-6. Cytokines induce matrix metalloproteinases (MMPs) and urokinase-type plasminogen activator (uPA). MMPs and uPA disrupt extracellular matrix. Since matrix damage may play a role in the neuropathological changes seen with TMT toxicity, we determined the effect of TMT on proteolytic enzyme production. Adult rats were injected with 8.0 mg TMT/kg. At different times after TMT injection, tissue samples from frontal lobe and hippocampus were assayed for MMPs and uPA, using gelatin-substrate and casein/plasminogen-substrate zymography. Gelatinase B (92 kDa type IV collagenase) production increased significantly in frontal lobe tissue at 24, 48 and 96 h, and in hippocampus at 48 h compared to saline-injected controls. Gelatinase A (72 kDa type IV collagenase) was significantly decreased at 12 and 24 h in frontal lobe compared to controls. Urokinase-type PA was significantly increased in hippocampus at 12 and 96 h, and in frontal lobe at 96 h compared to controls. Gelatinase B and uPA are up-regulated by TMT in frontal lobe and hippocampus, suggesting that they may contribute to the neuropathology of TMT. PMID- 9218630 TI - Postischemic expression of P-selectin immunoreactivity in rat brain. AB - Change of immunoreactive P-selectin was examined in rat brain after transient middle cerebral artery (MCA) occlusion (O) with anti-P-selectin monoclonal antibody using brain samples of sham control and after ischemia. Temporal, spatial, and cellular changes of immunohistochemical expressions of P-selectin were evaluated with rat brain sections at 2 and 8 h, 1, 3, and 7 days of reperfusion after 1 h of MCAO. Western blot showed a single band at molecular weight of 140 kDa for P-selectin after ischemia. P-selectin immunoreactivity was not normally present in rat brain sections. However, it was expressed mainly in the post-capillary venules of the cerebral cortex and caudate in the MCA territory with a peak at 8 h-1 day. The expression was diminished by 3 days of reperfusion. The present results indicate that P-selectin was expressed from an earlier stage of reperfusion in post-capillary venules, and the expression became maximum at the same time both in the cerebral cortex and caudate. PMID- 9218631 TI - Neural network models and the visual cortex: the missing link between orientation selectivity and the natural environment. AB - Orientation selectivity is a basic property of neurones in the visual cortex of higher vertebrates. Such neurones can be seen to act as 'feature detectors', which provide an efficient cortical representation of the outside world. More recently, the removal of correlations between the signals of cortical neurones has been suggested as suitable theoretical concept for explaining the development of receptive fields. Corresponding neural network simulations yielded oriented 'receptive field' structures resembling those observed by neurophysiologists. The findings suggest that the 'decorrelation approach' can provide a causal relationship between characteristics of the physical world and brain function. However, we were able to reveal a basic deficit of the decorrelation approach which we illustrate by the construction of two artificial 'worlds', a 'Gaussian' one and an 'orientation-only' one. We show that, according to the decorrelation approach, oriented environmental features would be neither necessary nor sufficient for the development of oriented receptive fields. Thus the link between environmental structure and cortical orientation selectivity still awaits a theoretical explanation. PMID- 9218632 TI - Subtyping of alpha1-adrenoceptors responsible for the contractile response in the rat corpus cavernosum. AB - The subtyping of alpha1-adrenoceptors responsible for mediating contraction in isolated corpus cavernosum of mature male Wistar rats was studied pharmacologically. Concentration-response studies of the cavernosal smooth muscle to three agonists: methoxamine, norepinephrine and octopamine showed that methoxamine exhibited the highest potency in inducing contractile response; the respective pD2 values were: 6.22, 5.83 and 5.38. In the presence of 2-(2,6 dimethoxyphenoxyethyl)aminomethyl-1,4-benzodioxane (WB4101), a specific antagonist for alpha1A-adrenoceptors, a parallel rightward shift of the concentration-response curve to methoxamine was observed. On the other hand, chloroethylclonidine (CEC) caused a rightward shift of the concentration-response curve to methoxamine with significant suppression of the maximum response. The pA2 value for WB4101 obtained from Schild plot was 9.03 +/- 0.06 with slope (95% CL) equal to 0.955 (1.088-0.832). In the absence of extracellular calcium ions, the methoxamine-induced contraction was reduced by 92%. Ca2(+)-Channel blockers, nifedipine 10(-6) M and diltazem 10(-6) M decreased the contractile response by 18 and 23%, respectively. The present findings suggest that alpha1A-adrenoceptors are responsible for the methoxamine-induced contraction of the rat cavernosal smooth muscle. PMID- 9218633 TI - Copper-zinc superoxide dismutase and isolectin B4 binding are markers for associative and transhemispheric diaschisis induced by focal ischemia in rat cortex. AB - Copper/zinc-superoxide dismutase (Cu/Zn-SOD) belongs to a class of enzymes, identified as essential and highly effective endogenous scavengers of cytotoxic oxygen radicals. These radicals contribute to postlesional neurotoxicity. In order to determine the superoxide-scavenging potential of regions affected by unilateral cortical photothrombosis, we studied the changes in the distribution of Cu/Zn-SOD and the appearance of activated microglia by immunohistochemistry and isolectin B4 binding. Four hours postlesion, Cu/Zn-SOD increased significantly within a homotopic area of the contralateral hemisphere and in ipsilateral thalamic nuclei, whereas isolectin B4-positive microglia were upregulated at days 5 and 7 postlesion within the same regions. The contralateral increase in the amount of the superoxide-scavenging Cu/Zn-SOD indicates that this enzyme is induced by a retrograde reaction carried through callosal connections. PMID- 9218634 TI - Evoked transmitter release at neuromuscular junctions in wild type and cysteine string protein null mutant larvae of Drosophila. AB - Cysteine string proteins (CSPs) are synaptic vesicle proteins thought to be involved in neurotransmitter release. To obtain more information about the function of these proteins motor nerve terminals of wild type and CSP null mutant Drosophila larvae were depolarized and excitatory postsynaptic currents (EPSCs) were recorded with an extracellular electrode at 16-18 degrees C. At this temperature the amplitude of average EPSCs was reduced and the time constant of the exponential fit of the current decay was increased in CSP null mutant compared to wild type larvae. The number of quanta released per pulse was not different but the time course of release was distributed differently in CSP null mutant and wild type larvae. In measurements of the latency of quantal EPSCs the probability of release after a pulse reached a lower peak value and the decay after the peak was delayed in CSP null mutant compared to wild type larvae. In addition facilitation in response to twin-pulse stimulation was slightly increased at low levels of release in CSP null mutant larvae. It is concluded that CSPs are involved in neurotransmitter release and help to synchronise evoked release at nerve terminals. PMID- 9218635 TI - Possible involvement of G-proteins in the regulation of striatal dopamine D2 receptor affinity by cholecystokinin octapeptide. AB - A G(i)-protein antibody AS/7 at 1:10 dilution significantly increased the K(d) values of the D2 agonist [3H]N-propylnorapomorphine (NPA) binding sites in the rat striatal membranes, and coincubation with sulphated cholecystokinin octapeptide (CCK-8; 1 nM) did not further increase the K(d) values. A GTP analogue guanylyl-imidodiphosphate (GMP-PNP) at 100 microM markedly increased the K(d) values of the [3H]NPA binding sites in the rat forebrain sections, and coincubation with CCK-8 (1 nM) again did not produce a further increase in the K(d) values. The present results indicate that abnormal activity of G-proteins abolished the ability of CCK-8 to reduce the D2 receptor affinity in the brain. PMID- 9218636 TI - Magnesium sulphate improves neurologic outcome following severe closed head injury in rats. AB - While recent evidence suggests that brain intracellular free magnesium concentration declines following severe diffuse traumatic brain injury, no studies have examined whether magnesium administration following such injury can improve subsequent neurologic outcome. The present study shows that MgSO4 administered as a bolus at 30 min following severe closed head injury in rats significantly improves posttraumatic neurologic outcome as assessed by both rotarod and angleboard tests. Moreover, this improvement in outcome was evident with both intravenous and intramuscular drug administration. We conclude that parenteral administration of magnesium sulphate may be neuroprotective following severe closed head injury of a diffuse nature. PMID- 9218637 TI - Geniculo-cortical afferents form synaptic contacts with vasoactive intestinal polypeptide (VIP) immunoreactive neurons of the rat visual cortex. AB - The lateral geniculate nucleus of the rat was injected with the anterograde tracer Phaseolus vulgaris leucoagglutinin (PHA-L) to see if geniculo-cortical axons terminate on vasoactive intestinal polypeptide immunoreactive (VIP-IR) neurons of the primary visual cortex. PHA-L-labelled boutons attached to VIP-IR perikarya and dendrites were identified as presynaptic parts of asymmetrical synapses. This geniculo-cortical projection to VIP-IR cells in the visual cortex and comparable findings in the somatosensory cortex suggest that sensory input from specific thalamic nuclei may influence local circuit inhibition and the metabolic state within the cortical domain via VIP-IR neurons. PMID- 9218638 TI - Brain functional activity during gait in normal subjects: a SPECT study. AB - The purpose of this study was to evaluate changes in brain activity during voluntary walking in normal subjects using technetium-99m-hexamethyl propyleneamine oxime single photon emission computed tomography. This study included 14 normal subjects. Statistical parametric mapping analysis revealed that the supplementary motor area, medial primary sensorimotor area, the striatum, the cerebellar vermis and the visual cortex were activated. These results suggested that the cerebral cortices controlling motor functions, visual cortex, basal ganglia and the cerebellum might be involved in the bipedal locomotor activities in humans. PMID- 9218639 TI - Attenuation of the catecholamine responses by electroacupuncture on Jen-Chung point during postoperative recovery period in humans. AB - In this study, Jen-Chung (J-C) point was stimulated by electroacupuncture (EA) in 10 patients, and by placebo treatment in 10 controls, immediately after termination of inhalation for 15 min. During the postoperative recovery period, plasma catecholamine (CA) levels were assessed before (0) and 15 and 30 min after treatment. The time from cessation of inhalation to the first eye opening and to extubation did not differ between groups. The plasma catecholamine levels increased by 30% from 0 to 15 min in the control group but decreased by 6% in the EA group. The levels at 30 min were approximately the same as at time 0. The change in catecholamine levels from 0 to 15 min was significantly lower (P < 0.02) in the EA groups than the control group. PMID- 9218640 TI - L-AP4 inhibition of depolarization-evoked cGMP formation in rat cerebellum. AB - The effects of the group III mGluR agonist, L-2-amino-4-phosphonobutyrate (L AP4), on depolarization-stimulated cGMP levels in adult rat cerebellar slices were determined. L-AP4 elicited a concentration-dependent, complete inhibition of cGMP formation stimulated by 4-aminopyridine (4-AP; 1 mM), yielding an IC50 value of 4.2 +/- 1.6 microM (n = 3). The 4-AP response was also reduced by the P-type Ca2+ channel toxins omega-conotoxin MVIIC (3 microM; 39 +/- 7% inhibition) and omega-Agatoxin IVA (30 nM; 53 +/- 4%), and was abolished in the absence of Ca2+ or in the presence of Co2+. The inhibitions of the 4-AP cGMP response by 10 microM L-AP4 and 30 nM omega-Agatoxin IVA were not additive, indicating that part of the actions of L-AP4 in the cerebellum involves the modulation of P-type Ca2+ channels. PMID- 9218641 TI - Immunohistochemical localization of calbindin D28k in the periodontal Ruffini endings of rat incisors. AB - It was examined whether calbindin D28k (CB) might be located in the rat incisor periodontal Ruffini ending, an essential mechanoreceptor in periodontal ligament, by light- and electron-microscopic immunohistochemistry. Some thick nerve fibers showing CB-like immunoreactivity (LI) entered the lingual half of the periodontal ligament of the incisor and showed the dendritic terminal arborization. Electron dense immunoreaction products indicating CB-LI were distributed diffusely in axoplasm of the axon terminals, no mitochondria, however, were not labeled. Neither cell bodies nor cytoplasmic extensions of the terminal Schwann cells exhibited CB-LI. CB was presumed to be involved in the maintenance of Ca2+ homeostasis in the mechano-electric transduction in mechanoreceptors in the periodontal ligament. PMID- 9218642 TI - Altered skin sensitivity in chronic itch: role of peripheral and central mechanisms. AB - Nodular prurigo (NP) is a chronic skin disease causing severe itch of unknown origin in restricted skin areas surrounded by healthy skin areas. In the present investigation we studied cutaneous sensibility in five NP-patients and in five control subjects. Pain thresholds were determined with short argon laser pulses using two different sizes of stimulus surface (diameters 2 and 4 mm), tactile threshold with calibrated monofilaments and skin blood flow with a laser Doppler flowmeter. We also studied the effect of prolonged capsaicin treatment which should predominantly impair the function of nociceptive C-fibers. In both the itching and healthy skin areas the pain thresholds were lower in NP-patients than in healthy control subjects. Before capsaicin, an increase in stimulus area produced an equal decrease in pain threshold in all subjects. Following prolonged capsaicin treatment the pain threshold obtained with a large but not a small stimulus surface was elevated to control levels in NP-patients. Tactile thresholds in NP-patients were lower than in control subjects, and this abnormality was reversed by capsaicin. The basal skin blood flow level was more labile (fluctuating) in itching skin areas than in healthy skin areas of NP patients. Capsaicin reduced blood flow fluctuation in the itching area. A lowered pain threshold not only in the itching area but also in the healthy skin area of NP-patients suggests that central convergence of itch and pain may contribute to increased pain sensitivity in chronic itch. Capsaicin-reversible abnormal fluctuation of the blood flow in the itching skin area might be explained by abnormal spontaneous activity of nociceptive peripheral nerve fibers and a consequent release of vasoactive agents from their terminals (axon reflex). The decreased tactile threshold and the elevation of it by capsaicin indicates that also the mechanisms underlying tactile sensibility are changed in chronic itch patients. PMID- 9218643 TI - Effect of stimulation of nicotinic cholinergic receptors on cortical cerebral blood flow and changes in the effect during aging in anesthetized rats. AB - The effect of intravenous injection of nicotine on cortical cerebral blood flow (CBF) was examined in urethane anesthetized rats. Nicotine (3-30 microg/kg) increased cortical CBF, independent of mean arterial pressure. This response was attenuated to about a half of the control one after lesioning the nucleus basalis of Meynert (NBM) bilaterally. The response was not significantly influenced after blocking the muscarinic receptors, but was abolished after blocking the nicotinic receptors in the parenchyma of the brain. It is concluded that the nicotine induced cortical vasodilation was mediated by activation of the nicotinic receptors in the NBM and also in the cortex of the brain. The threshold dose of nicotine for increasing cortical CBF was shifted in aged rats of 23-26 months, and the nicotine-induced increase in cortical CBF was much reduced in aged rats of 32-36 months. Activation of nicotinic receptors in the brain may be of therapeutic value in aged subjects in facilitating the cholinergic neural vasodilative system. PMID- 9218644 TI - Riluzole promotes survival of rat motoneurons in vitro by stimulating trophic activity produced by spinal astrocyte monolayers. AB - In the present study we have assessed whether riluzole stimulates the production of trophic activities for motoneurons by spinal astrocyte cultures. Astrocyte monolayers prepared from new-born rats were exposed to vehicle or riluzole (1-10 microM) for 30-36 h, then washed and further incubated without riluzole for 24 h in L15 medium to obtain the astrocyte conditioned media (ACM). Motoneuron enriched cultures were used to test the ability of the ACM to support motoneuron viability. Astrocyte monolayers exposed to 1 microM riluzole did not show changes in morphology or in DNA or protein synthesis. However, the conditioned medium obtained from astrocyte monolayers after this treatment increased motoneuron survival compared to that from vehicle-treated cultures. A similar effect was found when astrocytes were exposed to a higher riluzole concentration (10 microM) but with greater dilutions of the conditioned medium. This trophic activity was abolished by boiling or after treatment with trypsin. These findings strongly suggest the existence of a new trophic mechanism, through which riluzole may exert motoneuron protection. PMID- 9218646 TI - Fatal human plague--Arizona and Colorado, 1996. AB - In 1996, five cases of human plague, of which two were fatal, were reported in the United States; both decedents had septicemic plague that was not diagnosed until after they died. This report summarizes the investigation of the two fatal cases and underscores the need for health-care providers in areas with endemic plague to maintain a high level of awareness about the risk for plague in their patients. PMID- 9218645 TI - A polymorphism in the presenilin 1 gene does not modify risk for Alzheimer's disease in a cohort with sporadic early onset. AB - Mutations in the presenilin 1 gene account for many cases of early onset familial Alzheimer's disease. Homozygosity for the 'T' allele of a polymorphism in the presenilin 1 gene has previously been reported to double the risk for Alzheimer's disease in a late onset Caucasian sample. Here we report that this polymorphism does not incur risk in a case control sample of early onset Alzheimer's disease, possibly suggesting a different disease etiology between the early and late onset forms. PMID- 9218647 TI - Transmission of HIV possibly associated with exposure of mucous membrane to contaminated blood. AB - In February 1996, transmission of human immunodeficiency virus (HIV) by an unknown route involving an HIV-infected man and his previously uninfected female sex partner was reported to CDC. This report summarizes the epidemiologic investigation of this transmission, which suggests that the woman was infected through mucous membrane exposure to contaminated blood. PMID- 9218648 TI - Reduced susceptibility of Staphylococcus aureus to vancomycin--Japan, 1996. AB - Staphylococcus aureus is a virulent microorganism responsible for many serious infections among the general population. Since recognition of vancomycin resistant enterococci (VRE), the emergence of vancomycin resistance in S. aureus has been anticipated. This report describes the first documented case of infection caused by S. aureus with reduced susceptibility to vancomycin and includes the initial characterization of this isolate (1); the case occurred in a pediatric patient in Japan. The emergence of reduced vancomycin susceptibility in S. aureus increases the possibility that some strains will become fully resistant and that currently available antimicrobial agents will become ineffective for treating infections caused by such strains. PMID- 9218649 TI - Interim guidelines for prevention and control of Staphylococcal infection associated with reduced susceptibility to vancomycin. AB - Staphylococci are one of the most common causes of community- and hospital acquired infection. In many U.S. hospitals, strains of staphylococci (i.e., Staphylococcus aureus or coagulase-negative staphylococci) are resistant to all available antimicrobials except vancomycin. Rare cases of infection in the United States (1) have been caused by coagulase-negative staphylococci with reduced susceptibility to vancomycin (minimum inhibitory concentration [MIC] > or = 8 microg/mL)(2). PMID- 9218651 TI - Ecological monitoring helps researchers study disease in environmental context. PMID- 9218650 TI - Investigational treatments. How strict should we be? PMID- 9218652 TI - Psychiatrist explores apocalyptic violence in Heaven's Gate and Aum Shinrikyo cults. PMID- 9218653 TI - Belief in alien UFOs deep in American psyche. PMID- 9218654 TI - From the Centers for Disease Control and Prevention. Red blood cell transfusions contaminated with Yersinia enterocolitica--United States, 1991-1996, and initiation of a national study to detect bacteria-associated transfusion reactions. PMID- 9218655 TI - From the Centers for Disease Control and Prevention. Update: syringe-exchange programs--United States, 1996. PMID- 9218657 TI - Advantages and disadvantages of neonatal circumcision. PMID- 9218656 TI - A piece of my mind. Subject to payment? PMID- 9218658 TI - Advantages and disadvantages of neonatal circumcision. PMID- 9218659 TI - Advantages and disadvantages of neonatal circumcision. PMID- 9218660 TI - Advantages and disadvantages of neonatal circumcision. PMID- 9218661 TI - Advantages and disadvantages of neonatal circumcision. PMID- 9218662 TI - Advantages and disadvantages of neonatal circumcision. PMID- 9218663 TI - Advantages and disadvantages of neonatal circumcision. PMID- 9218664 TI - Should reimbursement be denied for liver transplantation in patients with hepatocellular carcinoma? PMID- 9218665 TI - Fifteen-year survival with prostate cancer in Sweden. PMID- 9218666 TI - Maternal infection and cerebral palsy in infants of normal birth weight. AB - CONTEXT: Exposure to maternal or placental infection is related to risk of preterm birth and, in premature infants, of brain lesions predictive of cerebral palsy (CP). Few studies have investigated whether maternal infection is associated with risk of CP in children of normal birth weight. OBJECTIVE: To investigate maternal infection during the admission for delivery as a possible risk factor for CP in infants born weighing 2500 g or more. DESIGN: Population based case-control study. SETTING: All hospitals in 4 northern California counties, 1983 through 1985. PARTICIPANTS: A total of 46 children with disabling spastic CP who had no recognized prenatal brain lesions and 378 randomly selected control children weighing 2500 g or more at birth and surviving to age 3 years. MAIN OUTCOME MEASURES: Disabling spastic CP and signs of neonatal morbidity. RESULTS: Maternal fever exceeding 38 degrees C in labor was associated with increased risk of unexplained CP (odds ratio [OR], 9.3; 95% confidence interval [CI], 2.7-31.0), as was a clinical diagnosis of chorioamnionitis. One or more indicators of maternal infection were present in 2.9% of control children, 22% of children with CP (OR, 9.3; 95% CI, 3.7-23.0), and 37% of those with the spastic quadriplegic subtype of CP (OR, 19.0; 95% CI, 6.5-56.0). Newborns exposed to maternal infection, both cases and controls, had 5-minute Apgar scores below 6 more often than those unexposed. Among children with CP, those born to infected women were more often hypotensive, needed intubation, had neonatal seizures, and received a clinical diagnosis of hypoxic-ischemic encephalopathy. CONCLUSION: Intrauterine exposure to maternal infection was associated with a marked increase in risk of CP in infants of normal birth weight. Maternal infection was also linked with low Apgar scores, other evidence of hypotension [corrected] and need for resuscitation, and neonatal seizures-signs commonly attributed to birth asphyxia. PMID- 9218667 TI - Prevention of heart failure by antihypertensive drug treatment in older persons with isolated systolic hypertension. SHEP Cooperative Research Group. AB - CONTEXT: Heart failure is often preceded by isolated systolic hypertension, but the effectiveness of antihypertensive treatment in preventing heart failure is not known. OBJECTIVE: To assess the effect of diuretic-based antihypertensive stepped-care treatment on the occurrence of heart failure in older persons with isolated systolic hypertension. DESIGN: Analysis of data from a multicenter, randomized, double-blind, placebo-controlled clinical trial. PARTICIPANTS: A total of 4736 persons aged 60 years and older with systolic blood pressure between 160 and 219 mm Hg and diastolic blood pressure below 90 mm Hg who participated in the Systolic Hypertension in the Elderly Program (SHEP). INTERVENTION: Stepped-care antihypertensive drug therapy, in which the step 1 drug is chlorthalidone (12.5-25 mg) or matching placebo, and the step 2 drug is atenolol (25-50 mg) or matching placebo. MAIN OUTCOME MEASURES: Fatal and nonfatal heart failure. RESULTS: During an average of 4.5 years of follow-up, fatal or nonfatal heart failure occurred in 55 of 2365 patients randomized to active therapy and 105 of the 2371 patients randomized to placebo (relative risk [RR], 0.51; 95% confidence interval [CI], 0.37-0.71; P<.001; number needed to treat to prevent 1 event [NNT], 48). Among patients with a history of or electrocardiographic evidence of prior myocardial infarction (MI), the RR was 0.19 (95% CI, 0.06-0.53; P=.002; NNT, 15). Older patients, men, and those with higher systolic blood pressure or a history of or electrocardiographic evidence of MI at baseline had higher risk of developing heart failure. CONCLUSION: In older persons with isolated systolic hypertension, stepped-care treatment based on low-dose chlorthalidone exerted a strong protective effect in preventing heart failure. Among patients with prior MI, an 80% risk reduction was observed. PMID- 9218668 TI - Relationship between National Institutes of Health research awards to US medical schools and managed care market penetration. AB - CONTEXT: Medical research conducted in academic medical centers is often dependent on support from clinical revenues generated in these institutions. Anecdotal evidence suggests that managed care has the potential to affect research conducted in academic medical centers by challenging these clinical revenues. OBJECTIVE: To examine whether empirical evidence supports a relationship between managed care and the ability of US medical schools to sustain biomedical research. DESIGN: Data on annual extramural research grants awarded to US medical schools by the National Institutes of Health (NIH) from fiscal years 1986 to 1995 were obtained, and each medical school was matched to a market for which information about health maintenance organization (HMO) penetration in 1995 was available. MAIN OUTCOME MEASURES: Growth in total NIH awards, traditional research project (R01) awards, R01 awards to clinical and basic science departments, and changes in institutional ranking by NIH awards were compared among schools located in markets with low, medium, and high managed care penetration. RESULTS: Medical schools in all markets had comparable rates of growth in NIH awards from 1986 to 1990. Thereafter, medical schools in markets with high managed care penetration had slower growth in the dollar amounts and numbers of NIH awards compared with schools in markets with low or medium managed care penetration. This slower growth for schools in high managed care markets was associated with loss of share of NIH awards, equal to $98 million in 1995, and lower institutional ranking by NIH awards. Much of this revenue loss can be explained by the slower growth of R01 awards to clinical departments in medical schools in high managed care markets. CONCLUSIONS: These findings provide evidence of an inverse relationship between growth in NIH awards during the past decade and managed care penetration among US medical schools. Whether this association is causal remains to be determined. PMID- 9218669 TI - Relationship between market competition and the activities and attitudes of medical school faculty. AB - CONTEXT: Growth in health care market competition and the concomitant increasing dependence of academic health centers on clinical revenues may require medical school faculty to increase patient care activity, perhaps at the expense of research and teaching. However, the relationship between health care market competitiveness and the activities and attitudes of medical school faculty has not been established. OBJECTIVE: To examine the relationship between market competitiveness and the activities and attitudes of medical school faculty members. DESIGN: Mailed survey of 3394 life-science faculty in the 50 universities that received the most funding from the National Institutes of Health in 1993. SETTING: Medical schools in research-intensive universities. PARTICIPANTS: A total of 2167 faculty responded to the survey (response rate, 64%). We analyzed the responses of 1671 medical school research faculty located in markets of differing health care competitiveness, ranging from least competitive (stage 1) to most competitive (stage 4) markets. MAIN OUTCOME MEASURES: The number of publications in refereed journals in the last 3 years, the average number of hours per week of teaching contact, whether faculty in clinical departments had patient care responsibilities, and measures of departmental community, cooperation, and conflict. RESULTS: Clinical researchers in stage 1 and 2 markets published more scientific articles than those in stage 3 markets (14.5 vs 12.6, P=.04) or in stage 4 markets (14.5 vs 12.0, P=.03). Among 96 young faculty (professional age-<10 years) in clinical departments, 11 (44%) of those in stage 1 and 2 markets had patient care duties compared with 32 (56.1%) of young faculty members in stage 3 markets (P=.04) and 12 (85.7%) of those in stage 4 markets (P=.01). The percentage of senior faculty in clinical departments (n=691) with patient care responsibilities did not differ significantly by market stage. Compared with faculty in less competitive markets, faculty in stage 4 markets perceived lower levels of departmental cooperation and higher levels of conflict. CONCLUSIONS: Increased competitiveness of health care markets seems to hinder the capacity of academic health centers to conduct clinical research and to foster the careers of young clinical faculty. PMID- 9218670 TI - Funding for patient-oriented research. Critical strain on a fundamental linchpin. AB - CONTEXT: Interest in clinical investigative careers has declined over the past 2 decades. While several factors are likely involved in this decline, one is the perceived difficulty in obtaining support for investigator-initiated clinical research projects. OBJECTIVE: To analyze the priority scores and funding rates of patient-oriented research (POR) compared with laboratory-oriented research (LOR) when grant applications to the National Institutes of Health (NIH) are reviewed by study sections of the NIH Division of Research Grants. DESIGN: Research grant applications submitted to NIH were classified by the applicant as involving human subjects or not (LOR). Those classified as involving human subjects were divided into clinical (POR) and nonclinical research. The association of priority score and POR or LOR status was evaluated using chi2 statistical techniques. SETTING AND PARTICIPANTS: Twelve thousand investigator-initiated grant applications (RO1s) in 2 of the 1994 NIH review cycles. MAIN OUTCOME MEASURES: Grant application priority scores and funding rates. RESULTS: On the basis of the following 3 criteria, POR applications fare less well than LOR applications: (1) POR status and ranking in the total application pool; (2) percentage of POR vs LOR applications in the top 20th percentile; and (3) funding rates of POR applications. Furthermore, the fate of a POR application depended on which study section reviewed the application. Those applications that were reviewed in study sections that primarily reviewed POR applications fared equivalently to LOR applications; in contrast, POR applications reviewed in study sections that primarily reviewed LOR applications encountered a less favorable fate. CONCLUSIONS: These objective data provide strong support to the clinical research community's concern that investigator-initiated POR applications are not reviewed equitably at the NIH. By restructuring the review process, fairness is likely to be restored. Without restructuring, the POR component of the medical research community may be critically damaged. PMID- 9218671 TI - Recurrent angiotensin-converting enzyme inhibitor--associated angioedema. AB - CONTEXT: Angiotensin-converting enzyme (ACE) inhibitors are associated with an increased risk of angioedema, but the risk of recurrent angioedema if treatment is continued is not known. OBJECTIVE: To test the hypothesis that the association between ACE inhibitor use and angioedema may not be recognized and to determine characteristics of angioedema associated with continued use of ACE inhibitors. DESIGN: Retrospective cohort study. SETTING: Tennessee Medicaid program. PATIENTS: Medicaid enrollees aged 15 years or older who used an ACE inhibitor and had a first documented episode of angioedema between 1986 and 1992 were followed up for recurrent episodes through June 1993. MEASUREMENTS AND MAIN RESULTS: We previously identified 82 patients with a first confirmed diagnosis of angioedema during 51 752 person-years of ACE inhibitor use in this population (1.6 per 1000 person-years). Among these 82 patients, there were 16 outpatient recurrences of angioedema among 13 patients during 189 patient-years of follow-up (8.5 per 100 patient-years). The rate of angioedema was much higher in users of ACE inhibitors with continued exposure (18.7 per 100 patient-years) than in those whose use of the drug was discontinued (1.8 per 100 patient-years) (P=.001). Review of the medical records for patients taking ACE inhibitors who had recurrent angioedema revealed that physicians attributed angioedema to a number of causes not related to ACE inhibitor use, even after multiple recurrences. CONCLUSION: Continuing use of ACE inhibitors in spite of angioedema results in a markedly increased rate of angioedema recurrence with serious morbidity. PMID- 9218673 TI - Preventing the extinction of the clinical research ecosystem. PMID- 9218672 TI - Epidemiology of sepsis syndrome in 8 academic medical centers. AB - CONTEXT: Sepsis syndrome is a leading cause of mortality in hospitalized patients. However, few studies have described the epidemiology of sepsis syndrome in a hospitalwide population. OBJECTIVE: To describe the epidemiology of sepsis syndrome in the tertiary care hospital setting. DESIGN: Prospective, multi institutional, observational study including 5-month follow-up. SETTING: Eight academic tertiary care centers. METHODS: Each center monitored a weighted random sample of intensive care unit (ICU) patients, non-ICU patients who had blood cultures drawn, and all patients who received a novel therapeutic agent or who died in an emergency department or ICU. Sepsis syndrome was defined as the presence of either a positive blood culture or the combination of fever, tachypnea, tachycardia, clinically suspected infection, and any 1 of 7 confirmatory criteria. Estimates of total cases expected annually were extrapolated from the number of cases, the period of observation, and the sampling fraction. RESULTS: From January 4, 1993, to April 2, 1994, 12759 patients were monitored and 1342 episodes of sepsis syndrome were documented. The extrapolated, weighted estimate of hospitalwide incidence (mean+/-95% confidence limit) of sepsis syndrome was 2.0+/-0.16 cases per 100 admissions, or 2.8+/-0.17 per 1000 patient-days. The unadjusted attack rate for sepsis syndrome between individual centers differed by as much as 3-fold, but after adjustment for institutional differences in organ transplant populations, variation from the expected number of cases was reduced to 2-fold and was not statistically significant overall. Patients in ICUs accounted for 59% of total extrapolated cases, non-ICU patients with positive blood cultures for 11%, and non-ICU patients with negative blood cultures for 30%. Septic shock was present at onset of sepsis syndrome in 25% of patients. Bloodstream infection was documented in 28%, with gram-positive organisms being the most frequent isolates. Mortality was 34% at 28 days and 45% at 5 months. CONCLUSIONS: Sepsis syndrome is common in academic hospitals, although the overall rates vary considerably with the patient population. A substantial fraction of cases occur outside ICUs. An understanding of the hospitalwide epidemiology of sepsis syndrome is vital for rational planning and treatment of hospitalized patients with sepsis syndrome, especially as new and expensive therapeutic agents become available. PMID- 9218674 TI - Some imperatives for clinical research. PMID- 9218675 TI - Amniotic fluid infection and cerebral palsy. Focus on the fetus. PMID- 9218676 TI - Important new findings in sepsis. PMID- 9218677 TI - Effect of K+ channel modulation on mouse feeding behaviour. AB - The K+ channel antagonists, glucose (100 microg per mouse i.c.v.), tetraethylammonium (1 microg per mouse i.c.v.) and apamin (1 ng per mouse i.c.v.), reduced food intake of mice comparably to the two anorectic drugs, amphetamine (10 microg per mouse i.c.v.) and cocaine (50 microg per mouse i.c.v.). Conversely, the K+ channel openers, minoxidil (5 microg per mouse i.c.v.) and pinacidil (10 microg per mouse i.c.v.), elicited an orectic effect of the same intensity as that induced by 2-deoxyglucose (200 microg per mouse i.c.v.), aurothioglucose (200 microg per mouse i.c.v.) and neuropeptide Y (0.5 microg per mouse i.c.v.). The antisense oligodeoxyribonucleotide (1-3 nmol per injection) to mKv1.1 gene produced, at 72 h, a dose-dependent increase in food intake. A quantitative reverse transcription-polymerase chain reaction (RT-PCR) study demonstrated a reduction in cerebral mRNA levels only in the antisense oligodeoxyribonucleotide-treated group, indicating the absence of a sequence independent action. Mice receiving the K+ channel modulators or antisense oligodeoxyribonucleotide had unmodified motor coordination and inspection activity as revealed, respectively, by the rotarod and hole-board tests. The integrity and functionality of central K+ channels appears, therefore, to be fundamental in the regulation of food intake by mice. PMID- 9218678 TI - Distribution of apomorphine enantiomers in plasma, brain tissue and striatal extracellular fluid. AB - Steady-state concentrations of apomorphine enantiomers were measured in the extracellular fluid collected from rat brain striatum by microdialysis. The free and total concentrations of both enantiomers were also measured in plasma as well as the total concentrations in different brain regions (striatum, cortex and cerebellum). We noticed no regional difference in the total concentrations of the two enantiomers. The extracellular concentrations were much lower, amounting to 8% for R(-)-apomorphine and 4% for S(+)-apomorphine, of the total brain tissue concentrations. The microdialysis samples contained 12 times more (R(-) apomorphine and 5 times more S(+)-apomorphine than the free apomorphine measured in plasma. The extracellular concentrations of R(-)-apomorphine (129 +/- 20 pmol/ml) were significantly higher (P = 0.001, n = 6), than those of S(+) apomorphine (70 +/- 10 pmol/ml). These results indicate that both enantiomers of apomorphine concentrate equally in brain cells, and that a stereoselective uptake system could operate for R(-)-apomorphine at the blood-brain barrier level. PMID- 9218679 TI - Intrathecally administered c-fos antisense oligodeoxynucleotide decreases formalin-induced nociceptive behavior in adult rats. AB - c-fos antisense strategy was applied as a pharmacological approach to characterize its dose-dependent role and reversibility in the reduction of formalin-induced hyperalgesia. Nociceptive behavioral responses (weighted score, flinching response, licking/biting) following formalin (50 microl 5%) injection were assessed in adult Wistar rats receiving different doses (50 nM, 250 nM) of intrathecally administered c-fos antisense oligodeoxynucleotides at different times prior to formalin injections. The treatments dose dependently decreased both Fos immunoreactivity expression in dorsal horn of rat lumbar spinal cord and all nociceptive measures in the tonic phase of the formalin test. c-Fos correlated well with weighted pain score and/or flinching responses, but not with licking/biting behavior. With the exception of a 48-120 h period required for licking/biting behavior to be restored to its normal status, the suppressive effect on c-fos expression and other nociceptive behaviors disappeared 48 h following c-fos antisense oligodeoxynucleotide treatment. The results suggest a pharmacological potential of c-fos antisense oligodeoxynucleotides in the central nervous system to block immediate-early genes and their resulting physiological consequence following noxious stimulus. PMID- 9218680 TI - Pharmacological studies on YM992, a novel antidepressant with selective serotonin re-uptake inhibitory and 5-HT2A receptor antagonistic activity. AB - YM992 ((S)-2-[[(7-fluoro-4-indanyl)oxy]methyl]morpholine monohydrochloride) is a novel compound that has selective serotonin (5-hydroxytryptamine, 5-HT) re-uptake inhibition and 5-HT2A receptor antagonistic activity in vivo. YM992, fluoxetine and citalopram showed 5-HT uptake inhibition activity in l-5-hydroxy-tryptophan (l-5-HTP)-treated mice. YM992 and trazodone attenuated 5-HT2A/2C receptor agonist induced head-twitches in mice, indicating that these drugs had 5-HT2A receptor antagonistic activity. YM992 and amitriptyline were highly active in the mouse tail suspension test. In contrast, fluoxetine and citalopram showed only a tendency to reduce the immobility time. Single treatment with YM992 as well as trazodone and fluoxetine ameliorated the learning deficit of olfactory bulbectomized rats, whereas citalopram and amitriptyline showed an ameliorative effect only after chronic treatment. Although YM992 has moderate affinity for alpha1-adrenoceptors, alpha1-adrenoceptor antagonism of YM992 in vivo was 10 times weaker than that of trazodone. These results demonstrate that YM992 has 5 HT uptake inhibition and 5-HT2A receptor antagonistic activity in vivo, and suggest that YM992 may be a novel antidepressant with high efficacy in clinical use. PMID- 9218681 TI - Effects of ginsenosides on maze performance and brain choline acetyltransferase activity in scopolamine-treated young rats and aged rats. AB - In young adult rats with scopolamine-induced cognitive impairment, choline acetyltransferase activity was increased in the medial septum, but not in the diagonal band, caudate and hippocampus, 30 min after the injection of ginsenosides Rg1 or Re. Rb1 and Rd had no effect on choline acetyltransferase activity. Aged rats showed a smaller number of initial correct responses in the radial-arm maze and a lower choline acetyltransferase activity in the medial septum, diagonal band, caudate and hippocampus than did young adult rats. Repeated i.p. injections of Rg1 increased the number of initial correct responses and the activity of choline acetyltransferase in the medial septum, but not in the diagonal band, caudate and hippocampus, in aged rats. These results suggest that Rg1 and Re may contribute the ameliorative effects through an increase of choline acetyltransferase activity in the medial septum. PMID- 9218682 TI - Tranilast inhibits contraction of rat aortic smooth muscle. AB - Recently, the anti-allergic drug tranilast has been shown to reduce the rate of coronary restenosis after percutaneous transluminal coronary angioplasty. In this study, we investigated the effect of tranilast on contraction of and Ca2+ movement in vascular smooth muscle. We measured the isometric force and fura-2 estimated intracellular Ca2+ concentrations ([Ca2+]i) of rat aortic strips. Exposure of aortic strips to tranilast (0-500 microM) dose-dependently inhibited endothelin-1-induced increases in tension and [Ca2+]i elevation of the strips. Similar inhibition by tranilast was observed in response to high K+ stimulation. These results suggest that tranilast inhibits the contraction of vascular smooth muscle by inhibiting Ca2+ mobilization, which might be related to its preventive effect on coronary restenosis after percutaneous transluminal coronary angioplasty. PMID- 9218683 TI - Flunarizine, an anti-migraine agent, impairs nitroxidergic nerve function in cerebral arteries. AB - Flunarizine is an anti-migraine agent that blocks the Ca2+ entry across cell membrane. In order to obtain a clue of mechanisms underlying the migraine headache, modifications by flunarizine of the response to nitric oxide (NO), a cerebral vasodilator and algogenic agent, derived from perivascular nerves were evaluated. Relaxations due to nerve stimulation by electrical pulses (5 Hz) and nicotine (10(-4) M) in canine cerebral arterial strips were attenuated by treatment with flunarizine dose-dependently, whereas the responses to exogenous NO (10(-7)-10(-6) M) and nitroprusside (10(-8)-10(-6) M) were unaffected. The inhibition by the Ca2+ entry blocker of the response to electrical nerve stimulation and nicotine was obtained in a concentration (10(-6) M) that did not significantly relax the arterial strips. NO derived from perivascular nerve may be one of the factors involved in the genesis of migraine attack, which is expected to be relieved by a reduction of neural NO synthase activity associated with a decreased Ca2+ influx by flunarizine during nerve activation. PMID- 9218684 TI - Characterization of subtype of alpha1-adrenoceptor mediating vasoconstriction in perfused rat hind limb. AB - The subtype of alpha1-adrenoceptor mediating the exogenous noradrenaline-induced vasopressor response in perfused rat hind limb was determined by functional measurements and radioligand binding assays. The potencies (pA2 values) of alpha1A-adrenoceptor-selective antagonists, RS-17053 (N-[2-(2-cyclopropylmethoxy phenoxy) ethyl]-5-chloro-alpha,alpha-dimethyl-1H-indole-3-ethanamine hydrochloride), WB 4101 (2-(2,6-dimethoxyphenoxyethyl) aminomethyl-1,4 benzodioxane), 5-methyl-urapidil, and the alpha1D-adrenoceptor-selective antagonist, BMY 7378 (8-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8 azaspirol[4.5]de cane-7,9-dione), to inhibit the noradrenaline-induced vasopressor response determined by Schild plot were 9.47 +/- 0.21, 9.48 +/- 0.19, 8.10 +/- 0.27 and 6.66 +/- 0.14, respectively, with no slope significantly different from unity. The affinities (K(i) values) of these antagonists were determined by displacement of 125I-BE 2254 (2-beta(4-hydroxyphenyl) ethylaminomethyl)-tetralone) binding from the cloned alpha1a-, alpha1b-, alpha1d adrenoceptor, stably expressed in human embryonic kidney (HEK) 293 cells. The pA2 values of the above antagonists correlated well with the binding K(i) values only for alphaIA-adrenoceptors (r = 0.93), but not for alpha1B-adrenoceptors (r = 0.51) and alpha1D-adrenoceptors (r = 0.13). The concentration-vasopressor response curve for noradrenaline was not significantly affected by pretreatment with 50 microM chloroethylclonidine for 30 min. The results suggest that only alpha1A-adrenoceptors mediate the noradrenaline-induced vasopressor response in perfused rat hind limb. PMID- 9218685 TI - Mechanisms for histamine H1 receptor-mediated vasodilation in isolated canine lingual arteries. AB - Histamine and selective histamine receptor subtype agonists' effects on isolated and perfused canine lingual arteries were investigated with the cannula insertion method. In preparations preconstricted with phenylephrine, histamine and a selective histamine H1 receptor agonist, 2-pyridylethylamine induced a biphasic vascular response in a dose-related manner, i.e., vasoconstriction followed by vasodilatation. The biphasic responses to histamine and 2-pyridylethylamine were inhibited by diphenhydramine, a selective histamine H1 receptor antagonist, but were not influenced by cimetidine, a selective histamine H2 receptor antagonist. Dimaprit, a selective histamine H2 receptor agonist, induced only a slight vasoconstriction which was not modified by cimetidine. Dimaprit never induced vasodilation even at a large dose. A histamine H3 receptor agonist, R-alpha methylhistamine, did not produce any significant vascular responses. Moreover, histamine-induced vasodilation was in part inhibited by removal of the endothelium, and the vasodilation remaining was abolished by H1 blockade. Thus, it is concluded that in canine lingual arteries there are abundant histamine H1 receptors which mediate both vasoconstriction and vasodilation, and that the histamine-induced vasodilation is in part due to endothelium-dependent mechanisms. PMID- 9218686 TI - Transcriptional regulated plasticity of vascular contractile endothelin ET(B) receptors after organ culture. AB - The aim of the present study was to investigate the level of regulation of the contractile endothelin ET(B) receptor which appears spontaneously after organ culture of vascular segments. Endothelin-1 elicited a strong contraction while the selective endothelin ET(B) receptor agonist, sarafotoxin 6c, had a negligible effect on fresh ring segments of rat mesenteric artery. After organ culture in serum-free Dulbecco's modified Eagle's medium at 37 degrees C (for 1 or 2 days) the endothelin-1-induced contraction was unchanged, whereas sarafotoxin 6c induced, after 1 day, a marked contraction which was further increased at day 2. The contraction induced by sarafotoxin 6c was significantly attenuated by the transcriptional inhibitor, actinomycin D, or the translational inhibitor, cyclohexamide, while the endothelin-1-induced contraction was much less affected. mRNA for endothelin ET(A) and endothelin ET(B) receptors was present in fresh human omental arteries denuded of endothelium. However, after organ culture, endothelin ET(B) mRNA was more prominent than endothelin ET(A) mRNA. Furthermore, the mRNA for both receptors was decreased after treatment with actinomycin D but not with cyclohexamide. This suggests that the endothelin ET(A) receptor is the dominating contractile receptor in fresh arteries while organ culture induces transcription and subsequent translation of contractile endothelin ET(B) receptors. PMID- 9218687 TI - Neuropeptide Y Y1 receptors are involved in the vasoconstriction caused by human sympathetic nerve stimulation. AB - Neuropeptide Y, a novel neurotransmitter, interacts with selective membrane receptors to cause vasoconstriction. Frequency- and concentration-dependent isometric contractions were observed in human inferior mesenteric artery and vein mounted rings that were stimulated with either electrical pulses (70 V, 0.5 ms, 2.5-20 Hz) or noradrenaline. The antagonism elicited by 100 nM tetrodotoxin and 1 microM guanethidine confirmed the neuronal and sympathetic origins of the vasomotor response. Incubation with BIBP 3226 ((R)-N2-(di-phenacetyl)-N-(4 hydroxyphenyl)-methyl-D-arginineam ide), a selective neuropeptide Y Y1 receptor antagonist, significantly reduced the vasoconstriction. The incomplete antagonist activity of BIBP 3226 tends to support the hypothesis of sympathetic co transmission involving neuropeptide Y, adenosine 5'-triphosphate and noradrenaline. These findings were confirmed in parallel studies using rat superior mesenteric artery and vein ring preparations. PMID- 9218688 TI - Role of N-terminal active sites of galanin in neurally evoked circular muscle contractions in the guinea-pig ileum. AB - Synthetic galanin-related peptides and several galanin-(1-15)ol analogs were used to examine structure-function relationships of the N-terminal active site of galanin in more detail, using the guinea-pig ileum. The synthetic peptides examined showed their inhibitory activity on the neurally evoked circular muscle contractions with the following order of potency: rat, human and tuna galanin, galanin-(1-15)ol and [D-Trp8]galanin-(1-15)ol > N alpha-acetylated galanin-(2 15)ol, [Ala6,D-Trp8]galanin-(1-15)ol, galanin-(1-15) > tuna galanin-(1-15), [D Ala8]galanin-(1-15)ol, N alpha-acetylated galanin-(1-15)ol. In contrast, [D Thr6]galanin-(1-15)ol, [D-Tyr9]galanin-(1-15)ol and [D-Trp9]galanin-(1-15)ol were ineffective and showed no antagonistic activities to galanin. These results suggest that the L-configuration at positions 6 and 9 seems to be important for the inhibitory action of galanin on the neurally evoked guinea-pig circular muscle contractions. PMID- 9218689 TI - Differential effect of codeine on coughs caused by mechanical stimulation of two different sites in the airway of guinea pigs. AB - We studied the difference in the effects of codeine on coughs caused by mechanical stimulation to the larynx and to the bifurcation of the trachea in lightly anaesthetized guinea pigs. Mechanical stimulation to the larynx or the bifurcation of trachea caused a stable cough response. The response was reproducible over 60 min, when stimulation was repeatedly applied at 20-min intervals. No significant difference was found between the amplitudes of the responses to mechanical stimulation of the larynx and of the tracheal bifurcation. Codeine, 10, 20 and 50 mg/kg, dose dependently depressed the coughs caused by larynx stimulation. The antitussive, however, failed to depress the cough caused by stimulation to the tracheal bifurcation, although a large dose, 50 mg/kg, significantly depressed the cough. In capsaicin-treated guinea pigs, codeine at 20 mg/kg significantly depressed the cough caused by stimulation to the tracheal bifurcation. The present results suggest that cough caused by mechanical stimulation to the larynx might be more sensitive to codeine treatment than cough caused by stimulation to the bifurcation of trachea. Furthermore, it is suggested that coughs caused by mechanical stimulation to both sites might consist of at least two components as regards their pharmacological nature. PMID- 9218690 TI - [3H]8-OH-DPAT labels a 5-HT site coupled to inhibition of phosphoinositide hydrolysis in the dorsal raphe. AB - The present study was undertaken to compare the properties of the [3H]8-OH-DPAT (8-hydroxy-2-(di-n-propylamino)tetralin) binding site in the dorsal raphe nucleus with the hippocampal 5-HT1A receptor. In both tissues inclusion of 1 mM Mg2+ enhanced specific [3H]8-OH-DPAT binding, while 1 mM GTP decreased radioligand binding. [3H]8-OH-DPAT appears to bind to a single population of binding sites in both the hippocampus and the dorsal raphe nucleus, although the K(d) for the radioligand at the dorsal raphe site was five times that observed at the hippocampal 5-HT1A receptor. Similarly, although 5-HT and selective 5-HT1A receptor ligands displayed high affinity for the [3H]8-OH-DPAT binding site in the dorsal raphe nucleus, the affinity at the dorsal raphe site was less than that observed at the hippocampal 5-HT1A receptor. 8-OH-DPAT inhibited forskolin stimulated adenylyl cyclase activity in the hippocampus, but did not alter enzyme activity in the dorsal raphe nucleus. Conversely, 8-OH-DPAT inhibited the accumulation of [3H]inositol phosphates in the dorsal raphe nucleus, but not in the hippocampus. An inhibition of phosphoinositide hydrolysis in the dorsal raphe nucleus also was found with the putative 5-HT1A receptor selective ligands, flesinoxan and gepirone. However, addition of another putative 5HT1A receptor selective ligand, buspirone, did not alter the generation of [3H]inositol phosphates, but blocked the inhibitory effect of 8-OH-DPAT on phosphoinositide hydrolysis. These studies demonstrate that the 8-OH-DPAT binding site in the dorsal raphe nucleus displays a binding profile which is similar to the hippocampal 5-HT1A receptor, but unlike this 5-HT1A receptor the binding site in the dorsal raphe nucleus is negatively coupled to phosphoinositide turnover. PMID- 9218691 TI - Neonatal and preweanling rats are able to express short-term behavioral sensitization to cocaine. AB - The present study assessed the ability of suckling rats to express short-term behavioral sensitization to cocaine prior to weaning. Rat pups, aged either 3, 5, 10, 12, 17 or 19 days at the beginning of the experiment, were placed in a chamber after daily injection with cocaine (7.5 or 15 mg/kg. i.p.) for either 2 or 4 consecutive days, and were tested for behavioral responsiveness to cocaine in the same chamber 24 h later (at either 7, 14 or 21 days of age). Such a short post-treatment interval was adopted, along with a consistent pairing of the testing context with the drug effect and a sensitive technique of behavioral measurement (video recording), in order to maximize the possibility of detecting any cocaine sensitization. Locomotion was sensitized at all ages, after both regimens in 14-day-old pups, but solely after 2 injections in 21- and 4 injections in 7-day-old pups. Sensitization was also expressed via behaviors specific to each age. Four cocaine injections augmented cocaine-induced uncoordinated movements of head, paws and body (horizontal activity) in 7-day-old pups, and mouth movements in 14-day-old pups. In 21-day-old pups, sensitization was dose- and regimen-dependently expressed via adult-like stereotyped head movements. In neonatal 7-day-old pups, cocaine sensitization was also visible as reductions in immobility (both injection regimens). Contrary to previous studies, these results indicate that, given the use of an appropriate methodology, short term sensitization to the motoric effects of cocaine can be expressed by suckling rats prior to weaning, even after relatively short regimens of daily injections. PMID- 9218692 TI - N-methyl-D-aspartic acid-induced penile erection and yawning: role of hypothalamic paraventricular nitric oxide. AB - A dose of N-methyl-D-aspartic acid (NMDA, 50 ng) that induces penile erection and yawning when injected into the paraventricular nucleus of the hypothalamus, increased the concentration of NO2- from 1.10 +/- 0.28 microM to 7.32 +/- 1.12 microM and of NO3 from 4.96 +/- 0.69 microM to 10.5 +/- 1.61 microM in the paraventricular dialysate obtained from male rats by in vivo microdialysis. NO2- concentration was not increased by (+/-)-alpha-(amino)-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA, 100 ng) or by trans-(+/-)-1-amino-1,3 cyclopentanedicarboxylic acid (ACPD) (100 ng), which were unable to induce these behavioral responses. N-Methyl-D-aspartic acid effect on NO2- concentration, penile erection and yawning was prevented by dizolcipine (MK-801) (10-100 ng) or by the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (20 microg), but not by the oxytocin receptor antagonist [d(CH2)5,Tyr(Me)2,Orn8]vasotocin (100 ng), or by the guanylate cyclase inhibitor methylene blue (20 microg) given in the paraventricular nucleus 15 min before N methyl-D-aspartic acid or by the dopamine receptor antagonist haloperidol (0.5 mg/kg) given intraperitoneally 30 min before N-methyl-D-aspartic acid. In contrast, the nitric oxide scavenger hemoglobin (20 microg) given in the paraventricular nucleus prevented N-methyl-D-aspartic acid-induced NO2- concentration increase, but was unable to prevent penile erection and yawning. The results suggest that N-methyl-D-aspartic acid induces penile erection and yawning by increasing nitric oxide synthase activity in the paraventricular nucleus of the hypothalamus, possibly in the cell bodies of oxytocinergic neurons projecting to extra-hypothalamic brain areas and mediating these behavioral responses. PMID- 9218693 TI - Effects of modulation of nitric oxide on acoustic startle responding and prepulse inhibition in rats. AB - The nitric oxide-arginine pathway is intimately connected to the release of dopamine and glutamate, two neurotransmitter systems that may be dysfunctional in schizophrenia. In addition, nitric oxide synthase inhibitors share several behavioral effects with the psychotomimetic drug, phencyclidine. Previous research has found that phencyclidine-like drugs disrupt prepulse inhibition of the acoustic startle response, an animal model of sensorimotor gating, an attentional process that is disrupted in certain neuropsychiatric disorders in humans (e.g., acute schizophrenia). The purpose of the present study was to examine the effects of nitric oxide modulators in this model. Following injection with a nitric oxide modulator or phencyclidine, rats were placed in startle chambers in which they were exposed to acoustic pulses presented alone or preceded by a prepulse. As in previous reports, phencyclidine disrupted prepulse inhibition at doses that did not affect startle during pulse alone trials. In contrast, the nitric oxide synthase inhibitors, N(G)-nitro-L-arginine (L-NOARG) and N(G)-nitro-L-arginine methyl ester (L-NAME), dose-dependently decreased startle during pulse alone trials, but neither drug affected prepulse inhibition. A nitric oxide precursor, L-arginine, produced similar results. Sodium nitroprusside (a nitric oxide releaser) and 7-nitroindazole (a third nitric oxide synthase inhibitor) did not affect startle amplitudes during pulse alone or prepulse + pulse trials. The present results suggest that modulation of nitric oxide synthesis or availability does not disrupt sensorimotor gating of the acoustic startle response and is probably not involved in mediation of this type of attentional deficit in humans. PMID- 9218694 TI - Repeated activation of neurotensin receptors sensitizes to the stimulant effect of amphetamine. AB - Effects of repeated intracerebroventricular microinjections of 18 nmol/10 microl of neurotensin, [D-Tyr11]neurotensin, or saline were tested on motor activity in different groups of rats. One week after the fourth central injection, sensitivity to the behavioral stimulant effect of amphetamine (1 mg/kg, i.p.) was tested. As previously reported, neurotensin attenuated motor activity while [D Tyr11]neurotensin when compared to saline produced an initial suppression followed by an excitation. Despite such different behavioral effects, both peptides produced sensitization to the stimulant effect of amphetamine. These results show that repeated activation of neurotensin receptors produces long lasting changes in responsiveness to a psychostimulant drug. PMID- 9218696 TI - Polydeoxyribonucleotide (defibrotide) protects against post-ischemic behavioral, electroencephalographic and neuronal damage in the gerbil. AB - The effectiveness of defibrotide, a single-stranded polydeoxyribonucleotide compound, in preventing damage caused by cerebral ischemia was studied. Global ischemia was induced in anesthetized gerbils by bilateral carotid artery occlusion for 10 min. Defibrotide (100 mg/kg) or saline was injected, i.v., immediately after reperfusion. The following parameters were evaluated simultaneously: (1) electroencephalographic (EEG) spectral power, recorded before, during and after the ischemic period; (2) body temperature, monitored with a rectal thermistor probe after reperfusion for 120 min; (3) spontaneous motility, evaluated through a photocell system and quantified in terms of total distance travelled in 30 min, 1 h after recirculation and at periods over 15 days; (4) mnemonic functions assessed by passive avoidance test from 3 to 15 days after ischemia; (5) histological examination, 7 days after reperfusion, counting CA1 hippocampal neuronal cells. The ischemia-induced complete flattening of spectral power was significantly reversed (P < 0.01) by post-ischemic treatment with defibrotide between 30 and 90 min after ischemia. A complete recovery of total EEG spectral power was seen in the defibrotide group at 6 h and the saline ischemic group at 1 day. Seven days after bilateral carotid occlusion, there was a significant decrease in spectral power (-70% +/- 6) together with a loss of the number of CA1 cells in the saline ischemic group (-64%). Treatment with defibrotide significantly protected against the decrease in spectral power (-30% +/- 7) and cell loss (-9%). Finally, the number of animals found to be protected against the ischemia-induced flattening was significantly larger for defibrotide treated gerbils than for saline-treated animals throughout the experiment except for the third day. Body temperature was significantly decreased only at 30 min after reperfusion in both ischemic and sham-operated groups. Defibrotide reduced ischemia-induced hypermotility but only 6 h after the insult. The ischemia induced impairment of memory was partially reversed within 3 days in the defibrotide-treated animals and fully reversed within 7 days in the defibrotide group and 15 days in the saline group. Our results demonstrate that defibrotide, even when administered after the post-ischemic period, possesses anti-ischemic properties. The mechanism by which defibrotide protects the ischemic reperfused brain is still largely unknown. However, a neuroprotection via adenosine A1 and A2 subtype receptor interaction can be put forward. PMID- 9218695 TI - Adenosine A2 receptors modulate haloperidol-induced catalepsy in rats. AB - The effect of adenosine A1 and A2 receptor agonists and antagonists was investigated on haloperidol-induced catalepsy in rats. Pretreatment (i.p.) with the non-selective adenosine receptor antagonist, theophylline, or the selective adenosine A2 receptor antagonist, 3,7-dimethyl-1-propargylxanthine (DMPX), significantly reversed haloperidol-induced catalepsy, whereas the selective adenosine A1 receptor antagonists, 8-phenyltheophylline and 8-cyclopentyl-1,3 dipropylxanthine produced no effect. Similar administration of the adenosine A2 receptor agonists, 5'-(N-cyclopropyl)-carboxamidoadenosine and 5'-N ethylcarboxamidoadenosine (NECA), and the mixed agonists with predominantly A1 site of action, N6-(2-phenylisopropyl) adenosine or 2-chloroadenosine, potentiated haloperidol-induced catalepsy. Higher doses of the adenosine agonists produced catalepsy when given alone. However, N6-cyclopentyladenosine, a highly selective adenosine A1 receptor agonist, was ineffective in these respects. The per se cataleptic effect of adenosine agonists was blocked by DMPX and the centrally acting anticholinergic agent, scopolamine. Scopolamine also attenuated the potentiation of haloperidol-induced catalepsy by adenosine agonists. Further, i.c.v. administration of NECA and DMPX produced a similar effect as that produced after their systemic administration. These findings demonstrate the differential influence of adenosine A1 and A2 receptors on haloperidol-induced catalepsy and support the hypothesis that the functional interaction between adenosine and dopamine mechanisms might occur through adenosine A2 receptors at the level of cholinergic neurons. The results suggest that adenosine A2, but not A1, receptor antagonists may be of potential use in the treatment of Parkinson's disease. PMID- 9218697 TI - Carbamazepine increases extracellular serotonin concentration: lack of antagonism by tetrodotoxin or zero Ca2+. AB - Carbamazepine administration causes large increases in extracellular serotonin concentration and dose-related anticonvulsant effects in genetically epilepsy prone rats (GEPRs). In order to determine the generality of the effect on serotonin, we determined the anticonvulsant ED50 for carbamazepine against maximal electroshock seizures in outbred, non-epileptic Sprague-Dawley rats. We then administered anticonvulsant carbamazepine doses to Sprague-Dawley rats and observed extracellular serotonin concentration in hippocampi by way of microdialysis. We found that administration of carbamazepine, either systemically or through the dialysis probe, resulted in significant and dose-related increases in extracellular serotonin concentration. Basal serotonin release was decreased by tetrodotoxin administration through the dialysis probe. Tetrodotoxin administration through the dialysis probe did not decrease the effect of systemically or focally administered carbamazepine on extracellular serotonin concentration. Similarly, elimination of Ca2+ from the dialysate did not alter the release of serotonin caused by carbamazepine. These findings suggest that the serotonin releasing effect of carbamazepine does not take place by exocytosis and does not require action potentials in the brain area in which the release takes place. Further they suggest that the effect is mediated by an action of carbamazepine directly on serotonergic nerve terminals. PMID- 9218698 TI - Long-term therapy with trimetazidine in cardiomyopathic Syrian hamster BIO 14:6. AB - The cardiomyopathic Syrian hamster (CMH) of the strain BIO 14:6 is a model for both cardiac and skeletal muscle abnormalities. It has reduced longevity and noticeable hypertrophy of the heart and liver. At 220 days, CMHs display a total Ca2+ overload, 1.3-1.8-fold normal and a cytosolic Ca2+ concentration 2-4-fold higher than normal. Long-term oral treatment (18 mg/kg per day) with trimetazidine (anti-ischaemic drug), from age 30 to 350 days, was more efficient than the standard Ca2+ blocker verapamil. Trimetazidine increased the median survival time of CMH by 57% and the hypertrophy disappeared. The total Ca2+ level in CMHs reverted to that of normal Syrian hamsters (F1B). The cytosolic Ca2+ overload was limited to a factor of approximately 2. Therefore, trimetazidine possesses anti-Ca2+ properties and is effective in increasing survival and decreasing the heart and liver hypertrophy of CMH. This suggests that trimetazidine may be valuable in the prevention of congestive heart failure of similar aetiology. PMID- 9218699 TI - Endothelin-1 induces vasoconstriction and nitric oxide release via endothelin ET(B) receptors in isolated perfused rat liver. AB - Endothelin-1 (0.1, 1 and 10 nM) induced a significant increase in portal pressure and nitric oxide (NO) release in the isolated rat liver. The endothelin ET(B) receptor agonist, IRL 1620 (Suc-[Glu9,Ala(11,15)]endothelin-1-(8-21)) (0.1, 1 and 10 nM) also elicited a marked increase in portal pressure and NO release. The potency of endothelin-1 was higher than that of IRL 1620. The endothelin ET(A) receptor antagonist, BQ-123 (cyclo(-D-Trp-D-Asp-Pro-D-Val-Leu)) (1 and 10 microM), had no effect on the endothelin-1-induced change in portal pressure and NO current. In contrast, the endothelin ET(B) receptor antagonist, BQ-788 (N-cis 2,6-dimethylpiperidinocarbonyl-L-gamma-methyl-leucyl-D-1-++ +methoxycarbonyltryptophanyl-D-norleucine) (1 and 10 nM), attenuated the endothelin-1-induced change in portal pressure and NO current. Administration of N(G)-monomethyl-L-arginine (L-NMMA), a NO synthase inhibitor, completely abolished the endothelin-1- or IRL 1620-induced NO release. L-NMMA enhanced the increase in portal pressure and decrease in O2 consumption caused by endothelin 1. These results indicated that endothelin ET(B) receptors mediate both vasoconstriction and NO release and that NO plays a significant role in stabilizing microcirculation in isolated perfused rat liver. PMID- 9218700 TI - Antithrombotic effects in a rat model of aspirin-insensitive arterial thrombosis of desethyl KBT-3022, the main active metabolite of a new antiplatelet agent, KBT 3022. AB - The antithrombotic effect of desethyl KBT-3022, which is the main active metabolite of the new antiplatelet agent, KBT-3022 (ethyl 2-[4,5-bis(4 methoxyphenyl)thiazol-2-yl] pyrrol-1-ylacetate; a cyclooxygenase inhibitor), was determined using a photochemically induced arterial thrombosis model in the rat femoral artery. Pretreatment with desethyl KBT-3022 (0.1, 0.3 and 1 mg/kg, i.v.) prolonged the time required to achieve thrombotic occlusion in the femoral artery and inhibited collagen-induced platelet aggregation in whole blood ex vivo, each in a dose-dependent manner. In all 6 rats used, particularly at the highest dose (1 mg/kg, i.v.) tested, cyclic variations in blood flow were hardly ever observed and complete cessation of blood flow did not occur during the 30-min observation time. BM-13505 (1, 3 and 10 mg/kg, i.v.), a thromboxane A2 receptor antagonist, also prolonged the time to occlusion, but cyclic variations in blood flow did occur. On the other hand, aspirin (10 and 30 mg/kg, i.v.) had little effect in terms of preventing thrombosis, although it inhibited collagen-induced platelet aggregation to the same extent as did desethyl KBT-3022. Desethyl KBT-3022 inhibited the thrombin-induced aggregation of washed platelets in a concentration dependent manner (1-40 microM), whereas aspirin and BM-13505 did not. These findings suggest that the potent antithrombotic effect of desethyl KBT-3022 may be attributable in part to its additional ability to inhibit thrombin-induced platelet aggregation. Accordingly, thromboxane A2 and thrombin may be important thrombotic mediators in this rat model. PMID- 9218701 TI - Effects of niflumic acid on alpha1-adrenoceptor-induced vasoconstriction in mesenteric artery in vitro and in vivo in two-kidney one-clip hypertensive rats. AB - The influence of niflumic acid (3 and 10 microM), a Cl- channel antagonist, on cirazoline-induced vasoconstriction in isolated perfused mesenteric artery (5 ml/min) from two-kidney one-clip (2K1C) hypertensive and sham normotensive rats was examined. In addition, the effect of a single i.v. bolus injection of niflumic acid (3 mg/kg) on cirazoline-mediated reduction in vascular conductance in superior mesenteric artery was determined in pentobarbital-anaesthetized hypertensive and normotensive rats. Bolus injections of cirazoline induced a dose dependent transient increase in the perfusion pressure in vitro. In the presence of niflumic acid, cirazoline-mediated vasoconstriction was significantly inhibited. Cirazoline-induced vasoconstriction in isolated mesenteric beds was also significantly inhibited following perfusion with Cl(-)-free buffer. Pre perfusion of mesenteric blood vessels with Cl(-)-free buffer resulted in a significantly greater inhibition of cirazoline-mediated vasoconstriction in sham normotensive rats than in hypertensive rats. We found that in Cl(-)-free buffer, cirazoline-mediated vasoconstriction could be further inhibited by niflumic acid. Intravenous infusion of cumulative doses of cirazoline in vivo caused a dose dependent decrease in superior mesenteric vascular conductance. Pretreatment with niflumic acid significantly impaired cirazoline-mediated decreases in vascular conductance. Our results indicate that chloride ions play an important role in alpha1-adrenoceptor-mediated vasoconstriction in mesenteric blood vessels. In addition, the contribution of chloride ions in alpha1-adrenoceptor-mediated vasoconstriction in blood vessels from hypertensive rats appears to be reduced. PMID- 9218702 TI - Ondansetron facilitates neuromuscular transmission in the guinea-pig ileum. AB - The effects of ondansetron on the neuromuscular function of the guinea-pig ileum were investigated in vitro. Ondansetron, but not tropisetron or MDL 72222 (1alpha H-3alpha-5alpha-H-tropan-3-yl-3,5-dichlo robenzoate), enhanced submaximal electrically induced contractions (EC50) = 1.3 x 10(-5) M). Desensitization with 5-hydroxytryptamine (1 x 10(-5) M) or 2-methyl-5-HT (1 x 10(-5) M) abolished this facilitatory response, which remained unaltered after desensitization with 5 methoxytryptamine (1 x 10(-5) M) or addition of tropisetron, MDL 72222, N-acetyl 5-hydroxytryptophyl-5-hydroxytryptophan, SB203186 (1-piperidinylethyl-1H-indole-3 carboxylate hydrochloride), pirenzepine or hexamethonium. At higher concentrations, ondansetron decreased the electrically induced contractions (EC50 = 1 x 10(-4) M); the inhibitory response was unaffected by (-)-naloxone (1 x 10( 6) M) or idazoxan (1 x 10(-6) M). We conclude that, in the guinea-pig ileum, ondansetron elicits a biphasic response: facilitation of neuromuscular transmission mediated by a serotonergic receptor distinct from the 5-HT3, 5-HT4 or putative 5-HT1P receptors, and an inhibitory response that does not involve opiate or alpha2-adrenoceptors. PMID- 9218704 TI - Guanylin-, heat-stable enterotoxin of Escherichia coli- and vasoactive intestinal peptide-induced water and ion secretion in the rat intestine in vivo. AB - The heat-stable enterotoxin of Escherichia coli binds to an intestinal receptor, guanylyl cyclase-C, and produces cGMP to induce diarrhea. Guanylin is an endogenous ligand of this receptor. In the present in vivo study, the intestinal water and ion secretion induced by mucosal application of 2 nmol/ml guanylin or 5 or 10 units/ml heat-stable enterotoxin into closed loops was compared in the rat. The characteristics of secretion induced by cAMP following intravenous perfusion of 1.2 nmol/100 g per h vasoactive intestinal peptide were compared to those induced by cGMP. Unidirectional Na+ and Cl- fluxes were estimated by addition of 22Na into the loop and i.v. injection of 36Cl. Guanylin induced less water and ion secretion than that produced by heat-stable enterotoxin in the colon, confirming the results of in vitro studies, and also in duodenum and ileum. The cAMP- or cGMP-mediated response had a similar pattern, i.e., an inhibition of Na+ absorption and an increase in anion secretion. PMID- 9218703 TI - Anti-obesity effect of MPV-1743 A III, a novel imidazoline derivative, in genetic obesity. AB - MPV-1743 A III ((+/-)-4-(5-fluoro-2,3-dihydro-1H-inden-2-yl)-1H-imidazole) is a novel imidazoline derivative. In this study, it was shown to bind with high affinity to alpha2-adrenoceptor subtypes alpha2A (IC50) = 0.66 +/- 0.06 nM), alpha2B (IC50) = 3.8 +/- 0.53 nM), alpha2C (IC50) = 3.1 +/- 0.61 nM) in the recombinant S115 cells and to alpha2D (IC50 = 0.94 +/- 0.10 nM) in the rat submandibular gland. MPV-1743 A III also showed remarkably high affinity to alpha1-adrenoceptors (IC50 = 150 +/- 12 nM) in the rat cerebral cortex and to imidazoline I2b-binding sites (IC50) = 150 +/- 5.0 nM) in the rat liver. The functional alpha2-adrenoceptor antagonistic effect of MPV-1743 A III was demonstrated by studying the ability of orally administered MPV-1743 A III to reverse and prevent the alpha2-adrenoceptor agonist detomidine-induced mydriasis in rat. The anti-obesity effect of MPV-1743 A III was investigated in genetically obese (fa/fa) Zucker rats in two different phases of obesity. Chronic treatment with MPV-1743 A III (0.3 3 mg/kg per day p.o. for 3 weeks) dose dependently decreased weight gain in early-phase obesity. In fully established obesity, GDP binding to mitochondria and expression of uncoupling protein mRNA were increased in brown adipose tissue by MPV-1743 A III indicating an activation of non shivering thermogenesis. The present study shows that MPV- 1743 A III has a modest anti-obesity effect in the genetic rodent model of obesity. The relative importance of alpha2- and alpha1-adrenoceptors and imidazoline I2b-binding sites in mediating the effects of MPV-1743 A III needs further evaluation. PMID- 9218705 TI - Existence of dopamine D1 receptor on the sympathetic nerve endings in the guinea pig vas deferens. AB - The effects of selective dopamine receptor agonists and antagonists on sympathetic neuromuscular transmission were investigated in the guinea-pig vas deferens in order to test for the presence of presynaptic dopamine receptors. A single-pulse field stimulus induced a rapid monophasic contraction which was strongly inhibited by alpha,beta-methylene ATP, a P2X purinoceptor desensitizing agent. The contraction was also inhibited by 5-bromo-N-(4,5-dihydro-1H-imidazol-2 yl)-6-quinoxalinamine (UK 14,304), a selective alpha2-adrenoceptor agonist. This inhibition was antagonized by idazoxan, an alpha2-adrenoceptor antagonist, but not by R(+)-7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzaz epine hydrochloride (SCH-23390), a dopamine D1 receptor antagonist. Furthermore, the contractions were inhibited in a dose-dependent manner by R(+)-1-phenyl 2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8-diol hydrochloride (SKF-38393) and (+/ )-6-chloro-7,8-dihydroxy-3-allyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-be nzazepine hydrobromide (SKF-82958), dopamine D1 receptor agonists, and the inhibition was antagonized by both SCH-23390 and idazoxan, but not by spiperone, a dopamine D2 receptor antagonist. The results suggest that dopamine D1 receptors are located on the sympathetic nerve endings of guinea-pig vas deferens. PMID- 9218706 TI - Induction of fatty acid synthesis by pravastatin sodium in rat liver and primary hepatocytes. AB - We examined the effect of pravastatin sodium (pravastatin), a 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitor, on fatty acid synthesis in rat liver. The repeated administration of pravastatin to rats at 250 mg/kg for 7 days led to a 2.8-fold increase in fatty acid synthesis in the liver. The diurnal change of fatty acid synthesis was not affected by the treatment. Hepatic fatty acid synthase activity was increased 3.2-fold, while acetyl-CoA carboxylase activity was not changed by the repeated administration of pravastatin. In rat hepatocytes, the incubation with 2 microg/ml pravastatin for 24 h increased fatty acid synthase activity 1.5-fold, as well as HMG-CoA reductase activity 2.8-fold. These results suggest that HMG-CoA reductase inhibitors might increase fatty acid synthesis in vivo through the induction of hepatic fatty acid synthase. PMID- 9218707 TI - Detachment of glycolytic enzymes from cytoskeleton of melanoma cells induced by calmodulin antagonists. AB - Glycolysis, which is the primary energy source in cancer cells, is known to be controlled by allosteric regulators, as well as by reversible binding of glycolytic enzymes to cytoskeleton. We have previously found that different calmodulin antagonists decrease the levels of allosteric activators of glycolysis, and reduce ATP content and cell viability in B16 melanoma cells. Here we report of a novel, additional, mechanism of action of calmodulin antagonists in melanoma cells. We show that these drugs cause a detachment of the glycolytic enzymes, phosphofructokinase (ATP: D-fructose-6-phosphate 1-phosphotransferase, EC 2.7.1.11) and aldolase (D-fructose-1,6-bisphosphate D-glyceraldehyde-3 phosphate-lyase, EC 4.1.2.13), from cytoskeleton of B16 melanoma cells. This effect was dose- and time-dependent, and preceded the decrease in cell viability. The detachment of glycolytic enzymes from cytoskeleton would reduce the provision of local ATP, in the vicinity of the cytoskeleton-membrane and would affect cytoskeleton structure. Since the cytoskeleton is being recognized as an important modulator of cell function, proliferation, differentiation and neoplasia, detachment of the glycolytic enzymes from cytoskeleton induced by calmodulin antagonists, as well as their reported inhibitory action on cell proliferation, make these drugs most promising agents in treatment of cancer. PMID- 9218708 TI - Substance P and epibatidine-evoked catecholamine release from fractionated chromaffin cells. AB - Bovine chromaffin cells were separated by density gradient centrifugation into subfractions enriched with either > 90% adrenaline- or 70-80% noradrenaline producing cells. Concentrations of epibatidine (an alkaloid with nicotinic receptor activity) as low as 10 nM released adrenaline and noradrenaline from both fractions of cells maintained as monolayer cultures. The maximal effect was evoked by 30 nM epibatidine and was comparable to that evoked by 10 microM nicotine. The catecholamine release from the noradrenaline fraction was 30-40% higher than from the adrenaline fraction. Initial exposure to 50 nM epibatidine reduced release induced by a second exposure to the drug. There was cross desensitization between epibatidine and nicotine. Substance P inhibited the epibatidine-evoked catecholamine release from both fractions by up to 85% (IC50 = 3-5 microM). The release of noradrenaline was inhibited more than that of adrenaline. In addition, substance P protected the chromaffin cells against desensitization of the nicotinic receptor by epibatidine. The C-terminal heptapeptide sequence of substance P was 10 x less active, two N-terminal sequences did not modulate the catecholamine release. PMID- 9218709 TI - A partial agonist model used in the allosteric modulation of the NMDA receptor. AB - We used a partial agonist model to understand further the allosteric modulation of D,L-(E)-2-amino4-propyl-5-phosphono-3-pentenoic acid ([3H]CGP-39653) binding by glycine, 1-hydroxy-3-amino-2-pyrrolidone (HA-966) and 5,7-dichlorokynurenic acid at the NMDA receptor. Binding of [3H]CGP-39653 was investigated in homogenates of cortex, hippocampus and cerebellum of adult rat. Glycine, HA-966 and 5,7-dichlorokynurenic acid maximally decreased the binding of 10 nM of [3H]CGP-39653 by approximately 50, 40 and 22%, respectively. Glycine, HA-966 and 5,7-dichlorokynurenic acid reduced [3H]CGP-39653 binding with IC50 values of 0.31, 11 and 0.044 microM, respectively. The decrease in [3H]CGP-39653 binding was due to a reduced affinity (Kd) and number of binding sites (Bmax) by all three drugs at concentrations where approximately maximum inhibition was observed. Glycine, HA-966 and 5,7-dichlorokynurenic acid lowered the Bmax by approximately 29, 16 and 10%, respectively, whereas the Kd values were increased by approximately 84, 44 and 32%, respectively, in cortex and hippocampus. There was no change in the binding of [3H]CGP-39653 in the cerebellum. The model used revealed that neither 5,7-dichlorokynurenic acid nor HA-966 had partial agonist characteristics in respect with the allosteric modulation of [3H]CGP-39653 binding. Furthermore, the results showed that brain regions have different pharmacological profiles which may depend on the NMDA receptor subunit composition. PMID- 9218710 TI - Cloning and analysis of MART-1/Melan-A human melanoma antigen promoter regions. AB - The MART-1/Melan-A human melanoma tumor antigen can be recognized by T lymphocytes and appears to be involved in tumor regression. To study the transcriptional regulation of this important gene, the 5' untranslated (UT) region of the MART-1/Melan-A gene was cloned and sequenced. Human melanoma cell lines were screened for MART-1/Melan-A mRNA expression. Primer extension and northern analysis were performed to confirm the mRNA size and start site. Several overlapping fragments of 5'UT were isolated from genomic DNA by polymerase chain reaction (PCR) from the previously described sequence for an additional 700 bp upstream. The fragments isolated (ranging from 838 bp to 160 bp in length) were used to drive luciferase reporter gene expression in melanoma and non-melanoma cell lines. Tissue-specific promoter activity was found in a 233-bp fragment of 5' UT with an average index of induction of 35 fold. The 233-bp MART-1/Melan-A promoter does not appear to have cytokine (IL-2, IL-4, IL-7, GM-CSF, TNF-alpha or IFN-gamma) responsive elements when studied in transient transfection assays. PMID- 9218711 TI - A novel cDNA from Drosophila encoding a protein with similarity to mammalian cysteine-rich secretory proteins, wasp venom antigen 5, and plant group 1 pathogenesis-related proteins. AB - The CAP protein family is made up of a group of secreted proteins that share sequence similarity. Members of this family are found in animals, plants, and fungi, and their shared sequence similarity suggests that members share a common, but as yet unknown, molecular function. As a first step in defining the function of CAP family proteins, an 878 bp partial cDNA encoding a novel member of the CAP family was cloned by the polymerase chain reaction (PCR) from total RNA of adult Drosophila. The cDNA contained the complete coding sequence for a protein 256 amino acids in length, as well as the complete 3' untranslated region (UTR) and a portion of the 5' UTR. The protein, named Antigen 5-related (Agr), was most similar in sequence to antigen 5 (Ag5), a CAP family member found in social wasps and ants. The corresponding Agr RNA is about 1 kb in length and is present at all stages of development, with highest levels observed in adults. Agr RNA is transcribed from a single gene that is located within region 12F of the X chromosome. The identification of Agr in Drosophila expands the number of known CAP family members to well over four dozen. Further studies of Agr and the gene which encodes this protein using the Drosophila model system may help provide important insight into the molecular functioning of this little known, but increasingly significant protein family. PMID- 9218712 TI - Nucleotide sequence of a Drosophila melanogaster cDNA encoding a calnexin homologue. AB - A cDNA which encodes a calnexin (Cnx)-like protein from Drosophila melanogaster has been characterized. The deduced amino acid sequence shares several regions of homology with Cnx from other sources with two conserved motifs each repeated four times. The gene was found to be transcribed in various tissues and at all developmental stages. We have mapped the gene at chromosomal position 99A and we have also mapped the related gene coding for Drosophila calreticulin at 85E. PMID- 9218713 TI - An improved GFP cloning cassette designed for prokaryotic transcriptional fusions. AB - A new gfp cloning cassette designed for prokaryotic transcriptional fusions has been constructed. This cassette consists of gfp (containing the S65T 'red-shift' [Heim et al. (1995) Nature 373, 663-664] and F64L 'protein solubility' [Cormack et al. (1996) Gene 173, 33-38] mutations) flanked by convenient restriction sites, a translational enhancer, and a consensus ribosome binding site with an optimized spacer region. gfp fusion strains containing this cassette demonstrate from 40- to 80-fold greater fluorescence intensity than wild-type gfp fusion strains. Additionally, this cassette confers sufficient fluorescence to recipient cells to be used in low copy-number plasmids, with promoters conferring low levels of transcription, and in bacterial taxa other than Escherichia, such as Pseudomonas. PMID- 9218714 TI - The uvsF gene region in Aspergillus nidulans codes for a protein with homology to DNA replication factor C. AB - The UV-sensitive mutant uvsF201 of Aspergillus nidulans shows increased spontaneous and UV-induced mutation and generally resembles mutants defective in nucleotide excision repair (NER). Fully-complementing uvsF clones were isolated from cosmid and cDNA libraries for sequencing. The uvsF gene is approximately 3.75 kb long and codes for a predicted polypeptide of 1092 amino acids (aa). Three small introns are clustered early in the coding region of the protein. A major part of the sequence shows homology to human, mouse and yeast RFC1 genes which code for the large subunit of the DNA replication factor C. The uvsF gene product may therefore function primarily in general DNA replication but in addition be required for the replication step of DNA repair. Extended sequencing of the uvsF gene region identified a second closely adjacent gene of unknown function which is divergently transcribed from a small (0.2 kb) intergenic promoter region. PMID- 9218715 TI - Cloning and characterization of mouse Dhm2 cDNA, a functional homolog of budding yeast SEP1. AB - We have isolated mouse Dhm2 cDNAs encoding a homolog of budding yeast SEP1, whose product is involved in many cellular processes including meiosis, cellular senescence, and telomere maintenance. The putative Dhm2 protein (Dhm2p), which consists of 1687 amino acids and whose molecular weight is 191,400, matches the size of Sep1p and shares extensive homology with Sep1p especially in their N terminal regions. A multicopy plasmid containing of the Dhm2 cDNA complements the slow growth phenotype, sporulation defect, and DNA recombination defect caused by the sep1 mutation in yeast, indicating that Dhm2 is a functional homolog of SEP1. Since Dhm1, another SEP1 homolog we reported previously, only partially compensates for the sep1 mutation, we conclude that Dhm2 is a true homolog of SEP1. Northern analysis revealed that 5.8 kb mRNA corresponding to Dhm2 open reading frame is produced highly in testis. These results strongly suggest that Dhm2p participates in gametogenesis in mouse. PMID- 9218716 TI - Cloning of a rat cDNA encoding retinal dehydrogenase isozyme type I and its expression in E. coli. AB - Peptides sequenced from the purified rat liver cytosolic retinal dehydrogenase P1 [Posch, K.C., Burns, R.D. and Napoli, J.L., 1992. Biosynthesis of all-trans retinoic acid from retinal: recognition of retinal bound to cellular retinol binding protein (type I) as substrate by a purified cytosolic dehydrogenase. J. Biol. Chem. 267, 19676-19682] were used to design oligonucleotides for cloning its cDNA. The deduced amino acid sequence of P1, now designated retinal dehydrogenase type I or RalDH(I), has close similarity with mouse AHD-2 and rat kidney aldehyde dehydrogenase, but is distinct from rat phenobarbital-inducible aldehyde dehydrogenase (PIADH), the presumed rat liver homolog of mouse AHD-2. Rat kidney (100%) and lung (88%) show relatively high mRNA levels of RalDH(I), liver (34%) and brain (22%) have moderate levels, and testis (8%) has low levels. Retinoid status affects RalDH(I) mRNA levels differently in different tissues. E. coli-expressed RalDH(I) exhibits allosteric kinetics for retinal with a Hill coefficient of 1.7, a K0.5 value of 1.4 microM and a Vmax of 52 nmol min(-1) mg( 1) protein. These data establish the cospecificity of P1 and RalDH(I), show that retinoid status affects expression of its mRNA in a tissue-dependent manner, and illustrate that aldehyde dehydrogenase isozymes with extensive homology can participate in different metabolic paths, e.g., RalDH vs. PIADH. PMID- 9218717 TI - The human immunoglobulin kappa locus on yeast artificial chromosomes (YACs). AB - The human immunoglobulin kappa locus is a duplicated structure. Contigs of 600 kb with 40 Vkappa genes and 440 kb with 36 Vkappa genes had been established for the Ckappa proximal (p) and distal (d) copies, respectively. In addition the human genome contains more than 24 dispersed Vkappa genes, called orphons. In the present study, 22 kappa-locus derived YACs were analyzed in detail, while 30 orphon-derived YACs were characterized only with respect to some parameters. The kappa-locus derived YACs allowed three gaps to be closed which previously could not be bridged by cosmid and phage lambda cloning. At the 5' side, the p contig was extended in the YACs by 50 kb and the d contig by 16 kb. At the 3'side, the d contig was extended by 11.5 kb. Beyond the 3' end of the d contig a new Vkappa gene was found, which is located, according to pulsed field gel electrophoresis (PFGE) experiments, at a distance of at least 140 kb from the last Vkappa gene of the contig. This Vkappa gene, which was termed Z0, occurred on three YACs, albeit at distances smaller than 140 kb; this was probably due to deletions in the YACs caused by abundant repetitive sequences at the borders of the locus. According to its sequence and to the restriction map of its surroundings, Z0 is an orphon gene of the so-called Z family, of which several members are known to be dispersed throughout the genome. The possibility that Z0 has been the parent of the other Z orphons is discussed. PMID- 9218718 TI - Isolation and characterization of IS1 circles. AB - Transposase encoded by insertion sequence IS1 is produced from two out-of-phase reading frames by translational frameshifting that occurs in a run of adenines. An IS1 mutant with a single adenine insertion in the run of adenines efficiently produces transposase, resulting in generation of miniplasmids by deletion for a region adjacent to IS1 from a plasmid carrying the IS1 mutant. Here, we found that besides miniplasmids, cells harboring the plasmid contained minicircles without the region required for replication. Cloning and DNA sequencing of the minicircles revealed that most of them were IS1 circles consisting of the entire IS1 sequence and a sequence, 5-9 bp in length, which intervenes between terminal inverted repeats, IRL and IRR, of IS1. Analysis of more IS1 circles isolated by polymerase chain reaction revealed that the intervening sequence was derived from the region flanking either IRL or IRR in the parental plasmid, suggesting that IS1 circles are generated by an excision event from the parental plasmid. The IS1 circles may be formed due to the cointegration reaction occurring within the parental plasmid carrying IS1. PMID- 9218719 TI - General purpose tagging vectors for fission yeast. AB - We have designed a series of vectors for use in the fission yeast Schizosaccharomyces pombe that allow fusion of any protein of interest to a triple HA epitope or a GST domain. The HA epitope may be placed at the N terminus or the C terminus under three different versions of the nmt1 promoter, to allow varying levels of gene expression. The GST tag may be placed at the N terminus or C terminus under control of a fully active nmt1 promoter. This family of vectors has compatible restriction sites and modular design, so that the protein under study may be exchanged easily between different plasmids. Using the Cdc19p protein as a test case, we have demonstrated that these plasmids can express functional tagged proteins in the fission yeast cell. PMID- 9218720 TI - A group 3 LEA cDNA of rice, responsive to abscisic acid, but not to jasmonic acid, shows variety-specific differences in salt stress response. AB - A cDNA clone oslea3, encoding a group 3 late-embryogenesis abundant (LEA) protein was isolated from roots of rice seedlings (Oryza sativa L.). The encoded OSLEA3 protein has previously been found to accumulate to higher levels in roots of two salt-tolerant compared to a salt-sensitive rice variety in response to abscisic acid (ABA) [Moons et al., 1995. Plant Physiol. 107, 177-186]. The OSLEA3 protein (Mr 20.5, pI 6.5) characteristically contains ten imperfect 11-mer amino acid repeats. Exogenous application of ABA and exposure to salt shock (150 mM NaCl) rapidly induces a de novo, abundant oslea3 transcript accumulation in seedling roots, whereas application of jasmonic acid (9 microM) does not induce oslea3 expression. The stress-induced oslea3 transcript gradually declined upon prolonged salt shock, as wilting-induced damage became irreversible. oslea3 expression was compared for the salt-tolerant variety Pokkali and the salt sensitive cultivar Taichung N1. Higher maximal mRNA levels were found in roots of the tolerant variety, also declining less rapidly upon sustained salt shock, concomitant with a delayed drop in shoot water content. DNA blot analysis indicated the existence of a small oslea3 gene family in rice with an equal gene number in both ecotypes. The results suggest that a differential regulation of oslea3 expression is an aspect of the varietal differences in salt stress tolerance. PMID- 9218721 TI - Sequence of the 3' end of the simian hemorrhagic fever virus genome. AB - SHFV is a member of a new virus family which includes the genus arterivirus. We have cloned and sequenced 6,314 nt from the 3' end of the SHFV genome. This sequence encompasses nine complete ORFs which is three additional ORFs as compared to the other arteriviruses. We have numbered these ORFs 2a, 2b, 3, 4, 5, 6, 7, 8 and 9. At the 5' end of this sequence is a partial ORF (ORF 1b) of 1590 nt and at the 3' end is a poly(A) tract preceded by a 76 nt noncoding region. The coding capacity for each of the SHFV ORFs as well as the potential mass, pI and number of N-linked glycosylation sites for each of the encoded peptides was determined. PMID- 9218722 TI - Cloning and characterization of rat p27Kip1, a cyclin-dependent kinase inhibitor. AB - Cyclin-dependent kinase (Cdk) inhibitors play significant roles in the cell cycle control of various biological phenomena. To characterize the role of Cdk inhibitors in rat cells, we isolated a cDNA encoding rat p27Kip1, a 27-kDa Cdk inhibitor. The 1.04-kb cDNA of rat p27 contained an open reading frame of 197 amino acids that shared high homology with mammalian p27 and significant homology with mammalian p21Cip1 and p57Kip2. p27 mRNA was detected in most rat tissues and cell lines. The levels of p27 protein expression were similar in rat cell lines transformed by E1A and in normal cells. Rat p27 was able to interact with Cdk 2/4 and cyclin A/D in rat cells, but the amounts of rat p27 in Cdk2 complexes were different between transformed cells and normal cells. Thus, the formation of stable complexes of rat p27 may be modulated by E1A. Rat p27 protein could inhibit the increased Cdk2-associated kinase activity in transformed rat cells. PMID- 9218723 TI - Molecular cloning of 5-hydroxytryptamine (5-HT) type 1 receptor genes from the Japanese puffer fish, Fugu rubripes. AB - To characterize the structure of Fugu G-protein coupled receptor family and its evolutionary divergence, we have cloned and sequenced the Fugu 5-HT type 1 receptor genes by Polymerase Chain Reaction (PCR) with degenerate primers followed by phage library screening. The analysis of the deduced amino acid sequences showed that F1A alpha and F1A beta have the highest homology to the human 5-HT1A receptor (71.5% and 63.7%, respectively). Another clone, F1D, showed highest (70.5%) homology to the human type 1D receptor. The amino acid residues that are important for ligand binding have been conserved in these Fugu genes. The phylogenetic tree analysis suggests that the duplication event of the Fugu type 1A receptor may have occurred after the divergence of Fugu and the tetrapod lineage. PMID- 9218724 TI - Porin from the halophilic species Ectothiorhodospira vacuolata: cloning, structure of the gene and comparison with other porins. AB - The gene coding for the anion-specific porin of the halophilic eubacterium Ectothiorhodospira (Ect.) vacuolata was cloned and sequenced, the first such gene so analyzed from a purple sulfur bacterium. It encodes a precursor protein consisting of 374 amino acid (aa)-residues including a signal peptide of 22-aa residues. Comparison with aa sequences of porins from several other members of the Proteobacteria revealed little homology. Only two regions showed local homology with the previously sequenced porins of Neisseria species, Comamonas acidovorans, Bordetella pertussis, Alcaligenes eutrophus, and Burkholderia cepacia. Genomic Southern blot hybridization studies were carried out with a probe derived from the 5' end of the gene coding for the porin of Ect. vacuolata. Two related species, Ect. haloalkaliphila and Ect. shaposhnikovii, exhibited a clear signal, while the extremely halophilic bacterium Halorhodospira (Hlr.) halophila (formerly Ect. halophila) did not show any cross-hybridization even at low stringency. This result is in good accordance with a recently proposed reassignment within the family Ectothiorhodospiraceae, which included the separation of the extremely halophilic species into the new genus Halorhodospira. PMID- 9218726 TI - Hot-spot mutations in the p53 gene of liver nodules induced in rats fed DL ethionine with a methyl-deficient diet. AB - Male F-344 rats were fed for 15 weeks a methyl-deficient L-amino acid defined diet containing 0.05% DL-ethionine. Nodules protruding from the surface of the liver were dissected free of surrounding tissue, and polyadenylated RNA isolated from the nodules was reverse transcribed. The region of the p53 gene comprising codons 120-290 was amplified by the polymerase chain reaction, and cDNAs were sequenced. Mutations were detected in nodules obtained from 7 of 12 rats. In all seven cases, the same two point mutations were present. The first was at the first base of codon 246 and consisted of a C-->T transition (C:G-->T:A, Arg- >Cys), while the second was at the second base of codon 247 and consisted of a G- >T transversion (G:C-->T:A, Arg-->Leu). It is concluded that the hepatocarcinogen ethionine induces specific hot-spot p53 gene mutations; this is in contrast to the mutations at various sites previously observed to occur in rats fed a hepatocarcinogenic methyl-deficient diet alone. The results also provide the first evidence that ethionine is mutagenic in the rat. PMID- 9218725 TI - Li-Fraumeni syndrome--a molecular and clinical review. PMID- 9218727 TI - Gossypol inhibition of mitosis, cyclin D1 and Rb protein in human mammary cancer cells and cyclin-D1 transfected human fibrosarcoma cells. AB - The antiproliferative effects of gossypol on human MCF-7 mammary cancer cells and cyclin D1-transfected HT-1060 human fibrosarcoma cells were investigated by cell cycle analysis and effects on the cell cycle regulatory proteins Rb and cyclin D1. Flow cytometry of MCF-7 cells at 24 h indicated that 10 microM gossypol inhibited DNA synthesis by producing a G1/S block. Western blot analysis using anti-human Rb antibodies and anti-human cyclin D1 antibodies in MCF-7 cells and high- and low-expression cyclin D1-transfected fibrosarcoma cells indicated that, after 6 h exposure, gossypol decreased the expression levels of these proteins in a dose-dependent manner. Gossypol also decreased the ratio of phosphorylated to unphosphorylated Rb protein in human mammary cancer and fibrosarcoma cell lines. Gossypol (10 microM) treated also decreased cyclin D1-associated kinase activity on histone H1 used as a substrate in MCF-7 cells. These results suggest that gossypol might suppress growth by modulating the expression of cell cycle regulatory proteins Rb and cyclin D1 and the phosphorylation of Rb protein. PMID- 9218728 TI - A quantitative reverse transcriptase polymerase chain reaction-based assay to detect carcinoma cells in peripheral blood. AB - The presence of tumour cells in the circulation may predict disease recurrence and metastasis. To improve on existing methods of cytological or immunocytological detection, we have developed a sensitive and quantitative technique for the detection of carcinoma cells in blood, using the reverse transcriptase polymerase chain reaction (RT-PCR) identifying transcripts of the pancarcinoma-associated tumour marker EGP-2 (KSA or 17-1A antigen). The amount of EGP2 mRNA was quantified using an internal recombinant competitor RNA standard with known concentration and which is both reversely transcribed and co-amplified in the same reaction, allowing for a reliable assessment of the initial amount of EGP2 mRNA in the sample. Calibration studies, seeding blood with MCF-7 breast carcinoma cells, showed that the assay can detect ten tumour cells among 1.0 x 10(6) leucocytes. The PCR assay revealed that normal bone marrow expresses low levels of EGP2 mRNA, although immunocytochemistry with the anti-EGP2 MAb MOC31 could not identify any positively stained cell. Analyses using this RT-PCR assay may prove to have applications to the assessment of circulating tumour cells in clinical samples. PMID- 9218729 TI - Clinical relevance of bcl-2(MBR)/J(H) rearrangement detected by polymerase chain reaction in the peripheral blood of patients with follicular lymphoma. AB - We evaluated the prognostic role of peripheral blood polymerase chain reaction (PCR) assay for detection of the bcl-2(MBR)/J(H) rearrangement in 59 patients with follicular lymphoma (FL) treated at our centre since 1989. Thirty-five (59%) patients were bcl-2/J(H) positive and 24 (41%) were negative in the peripheral blood at diagnosis. Peripheral blood bcl-2/J(H) rearrangement detection at diagnosis had no relation to overall survival (OS) and time to progression (TTP). Peripheral blood PCR assay was performed post treatment in 17 patients who were bcl-2/J(H) positive at diagnosis. Fourteen of the patients (82%, 95% CI 56-96%) became bcl-2/J(H) negative. Nine of these patients were further analysed during follow-up and, after several months, circulating cells carrying the bcl-2/J(H) rearrangement reappeared in five of the nine patients. Peripheral blood clearance of bcl-2/J(H)-positive cells was correlated with better overall survival (log rank P < 0.05) but not with TTP. Our data confirmed that bcl-2(MBR)/J(H) rearrangement detection by PCR at diagnosis is not a prognostic factor in follicular lymphoma. In our series, clearance of circulating bcl-2/J(H)-positive cells appeared to correlate with better overall survival. Post-treatment examination of the peripheral blood by PCR may have clinical relevance for prediction of the survival pattern of the patients. PMID- 9218730 TI - Is a histological section representative of whole tumour vascularity in breast cancer? AB - The assessment of a tumour's angiogenic potential, by measuring the microvessel density in histological sections, assumes that a 4-microm section is representative of whole tumour vascularity. This study has examined this assumption by comparing the vessel density found radiologically, after injecting specimens with contrast, with that found immunohistochemically. Twenty-one breast angiograms were performed following mastectomy for carcinoma and graded 1-3 for vessel density. Sections (4 microm) from these carcinomas were labelled for endothelial cells using anti-CD34, and the vessel counts were compared with the radiological grades. A significant correlation was found between the densities (P < 0.003, Kruskal-Wallis one-way ANOVA). We therefore conclude that the microvessel density measured in histological sections is representative of whole tumour vascularity. PMID- 9218732 TI - Expression of high-affinity 67-kDa laminin receptors in primary breast cancers and metachronous metastatic lesions or contralateral cancers. AB - The presence of high-affinity 67-kDa laminin receptors, detected immunohistochemically, was determined on 63 primary breast cancers and on metachronous metastatic lesions or contralateral cancers from the same patients. A disagreement was observed in two-thirds of the cases. In particular, laminin receptor content was significantly lower (P = 0.02) in local recurrences and slightly higher in lymph node metastasis than in the corresponding primary tumours. PMID- 9218731 TI - p53 exon 5 mutations as a prognostic indicator of shortened survival in non-small cell lung cancer. AB - Inactivation of the tumour-suppressor gene p53 has been described as one of the most common molecular changes found in lung tumours. Our purpose was to study the prognostic value of p53 alterations and to determine whether some specific mutation type in the p53 gene could be associated with poor clinical evolution in non-small-cell lung cancer (NSCLC) patients. To this end, we studied 81 resected primary NSCLCs in order to detect p53 alterations. p53 protein accumulation was analysed using immunohistochemistry methods; p53 gene mutations in exons 5-9 were studied using polymerase chain reaction-single-strand conformation polymorphism and sequencing techniques. p53 protein was immunodetected in 46.9% of lung carcinomas and 44.7% of p53-immunopositive tumours showed p53 mutations. Survival analysis was performed on 62 patients. No survival differences were found for patients with or without p53 immunopositivity. A shorter survival was found in patients with underlying p53 gene mutations, mainly in patients with squamous cell lung tumours; the worst prognosis was found when mutations were located in exon 5 (P = 0.007). In conclusion, the location of p53 mutations might be considered as a prognostic indicator for the evaluation of poor clinical evolution in NSCLC patients. PMID- 9218733 TI - Prognostic significance of Ki-67 antigen and p53 protein expression in pancreatic duct carcinoma: a study of the monoclonal antibodies MIB-1 and DO-7 in formalin fixed paraffin-embedded tumour material. AB - Formalin-fixed paraffin-embedded material from 57 patients in whom curative resection of pancreatic carcinoma had been attempted was analysed by an immunohistochemical procedure to estimate proliferation and p53 protein expression. Using the monoclonal antibody (MAb) MIB-1, which recognizes a Ki-67 epitope, the proliferating cell index (PCI, percentage of immunoreactive tumour nuclei) and proliferating cell area (PCA, percentage of immunoreactive tumour nuclear area) were calculated using an interactive image analysis system and were compared with semiquantitative scoring of stainability. MAb DO-7, which recognizes both wild- and mutant-type p53 protein, was used to assess p53 expression in the same material. MIB-1 stainings were of high quality in 53 tumours. The median PCI was 29.7% (range 0.5-82.1%) and the median PCA was 10.6% (range 0.0-36.5%). There was a close correlation between PCI and PCA (P < 0.0001). PCI and PCA values were in conformity with the semiquantitative scoring (P < 0.0001). The p53 immunohistochemical stainings were successful in 48 tumours and the protein was expressed in 22 (46%). High PCI values (> 45%, n = 14) correlated with shorter survival time (P < 0.01). PCA (P < 0.05) and the expression of p53 protein (P < 0.001) were independent prognostic variables. PMID- 9218734 TI - Insulin-like growth factors and their binding proteins in human colonocytes: preferential degradation of insulin-like growth factor binding protein 2 in colonic cancers. AB - We have compared the expression of insulin-like growth factors (IGFs) and IGF binding proteins (IGFBPs) in ten paired samples of normal and tumour colonic tissue with regard to both mRNA and protein. We have compared sensitivity of these tissues to IGF-I using primary cultures of epithelial cells of colonic mucosa, and we have examined the production of IGFs and IGFBPs by these cells. In the tissues, IGFBP-2 mRNA was expressed in all normal and cancer samples but other IGFBPs showed variable expression. mRNAs for IGF-I were expressed in all normal and cancer tissues but IGF-II mRNA was only detected in cancer tissue (3 out of 10). Immunostaining of sections of normal and cancer tissue was negative for IGF-I and IGF-II; IGFBP-2 was positive in 2 out of 10 cancer tissues and 7 out of 10 normal tissues; IGFBP-3 was positive in 7 out of 10 cancer tissues and 7 out of 10 normal tissues; and IGFBP-4 was positive in 5 out of 10 cancer tissues and 6 out of 10 normal tissues. In the cells in culture, cancer cells showed increased incorporation of [35S]methionine into protein and [3H]thymidine into DNA (P < 0.02) when treated with IGF-I. Western blotting of serum-free conditioned media from cells in culture showed that 8 out of 10 normal and 3 out of 10 cancer cultures produced a 32-kDa immunoreactive IGFBP-2. No IGFBP-3 was secreted by any culture but 24-kDa IGFBP-4 was found in 3 out of 10 normal and 5 out of 10 cancer tissues. Because of the discrepancy between mRNA and protein expression for IGFBP-2, degradation of native IGFBPs was assessed using tissue extracts. Colon cancer extracts were able to degrade exogenous IGFBP-2, IGFBP-3 and IGFBP-4, whereas normal tissue extracts were without effect on IGFBP-2. We conclude that IGFBPs are synthesized and secreted by cells of the colonic mucosa but that proteolysis of secreted IGFBP-2 occurs in colon cancer tissue. This selective degradation may confer a growth advantage. PMID- 9218735 TI - Characterization of a clonal human colon adenocarcinoma line intrinsically resistant to doxorubicin. AB - Intrinsic low-level resistance to anti-cancer drugs is a major problem in the treatment of gastrointestinal malignancies. To address the problem presented by intrinsically resistant tumours, we have isolated two monoclonal lines from LoVo human colon adenocarcinoma cells: LoVo/C7, which is intrinsically resistant to doxorubicin (DOX); and LoVo/C5, which shows the same resistance index for DOX as the mixed parental cell population. For comparison, we have included in the study a LoVo-resistant line selected by continuous exposure to DOX and expressing a typical multidrug resistant (MDR) phenotype. In these cell lines we have studied the expression and/or activity of a number of proteins, including P-glycoprotein 170 (P-gp), multidrug resistance-associated protein (MRP), lung resistance related protein (LRP), glutathione (GSH)-dependent enzymes and protein kinase C (PKC) isoforms, which have been implicated in anti-cancer drug resistance. Intracellular DOX distribution has been assessed by confocal microscopy. The results of the present study indicate that resistance in LoVo/C7 cells cannot be attributed to alterations in P-gp, LRP or GSH/GSH-dependent enzyme levels. Increased expression of MRP, accompanied by alterations in the subcellular distribution of DOX, has been observed in LoVo/C7 cells; changes in PKC isoform pattern have been detected in both intrinsically and pharmacologically resistant cells. PMID- 9218736 TI - Relationship between folate-binding protein expression and cisplatin sensitivity in ovarian carcinoma cell lines. AB - It has been suggested that sensitivity of ovarian carcinomas to cisplatin is in part related to an endogenous folate deficiency. In this work, we investigated whether overexpression of the folate-binding protein (FBP), a receptor involved in folate transport, might be associated with cisplatin sensitivity. The results obtained on a panel of ten ovarian carcinoma cell lines that overexpress different levels of the FBP showed a statistically significant relationship between FBP overexpression and cisplatin responsiveness, with the most sensitive cell lines expressing higher FBP levels on their membrane than the less sensitive ones. The relationship was observed both in cells growing in standard medium containing high-folate concentrations (2.3 microM) and in cells adapted to growth in low-folate (20 nM) medium. Analysis of two cisplatin-resistant cell lines derived from the cisplatin-sensitive IGROV1 ovarian carcinoma cell line indicated that resistance was associated with a significant decrease in FBP expression. However, the receptor does not appear to be directly responsible for drug sensitivity per se as different cell lines transfected with FBP cDNA did not become more sensitive to the drug. Together, the data suggest the possible predictive value of FBP in ovarian carcinoma, as higher levels of expression can be indirectly but significantly associated with increased drug sensitivity. PMID- 9218737 TI - Modulation of doxorubicin resistance in a doxorubicin-resistant human leukaemia cell by an immunoliposome targeting transferring receptor. AB - Using a doxorubicin-resistant subline (K562/ADM) of human leukaemia K562 cells (Tsuruo et al, 1986), the effect of immunoliposomes that targeted a cellular transferrin receptor (TFR) was examined by neutralization of doxorubicin (DOX) resistance. OKT9-CIL, prepared by conjugation of DOX-encapsulated liposome with an anti-TFR monoclonal antibody, OKT9 (Aisenberg and Wilkes, 1980), showed similar binding to both K562 and K562/ADM. Although an 80-fold higher sensitivity to free DOX on cell growth inhibition in K562 than in K562/ADM was found, the difference was clearly diminished after OKT9-CIL treatment through the increased sensitivity of K562/ADM. The cellular DOX level 30 min after the exposure of free DOX was 45-fold lower in K562/ADM than in K562, whereas nearly equivalent DOX levels were detected in K562 and K562/ADM after OKT9-CIL treatment. In addition, DOX in K562/ADM in the free DOX treatment was efficiently excreted by 54% within 120 min of incubation, whereas almost all DOX supplied by OKT9-CIL remained uncleared. Fluorescence microscopic observation showed that OKT9-CIL was internalized into juxtanuclear vesicles in K562/ADM cells. These results suggest that OKT9-CIL has a potency to accumulate DOX, resulting in augmentation of DOX cytotoxicity in DOX-resistant tumour cells. PMID- 9218738 TI - Phase II study of high-dose dexamethasone-based association in acute and delayed high-dose cisplatin-induced emesis--JCOG study 9413. AB - Thirty-three patients with lung cancer receiving 80 mg m(-2) cisplatin were treated with high-dose dexamethasone (32 mg m(-2) on days 1-3, 16 mg m(-2) on day 4 and 8 mg m(-2) on day 5) combined with granisetron on day 1 and metoclopramide on days 2-5. Twenty-eight (85%) patients had no nausea or vomiting on day 1, and 16 (48%) achieved total control on days 1-5 with acceptable toxicity. High-dose dexamethasone for cisplatin-induced delayed emesis should be further evaluated in a phase III trial. PMID- 9218739 TI - Epirubicin combined with estramustine phosphate in hormone-resistant prostate cancer: a phase II study. AB - Twenty-four assessable patients with hormone-resistant prostate cancer (HRPC) were to receive daily doses of oral estramustine phosphate (EMP), 10 mg kg(-1), and intravenous epirubicin (EPR) infusions, 100 mg m(-2), every third week up to a cumulative dose of 500 mg m(-2). Biochemical response [> or = 50% reduction in pretreatment serum prostate-specific antigen (PSA) after three cycles of > or = 3 weeks' duration] was demonstrated in 13 of 24 patients included (54%). No objective response (WHO criteria) was observed, although seven of nine evaluable patients achieved a > or = 50% serum PSA reduction. Subjective improvement (pain score, performance status) occurred in 7 of 24 patients, whereas nine patients progressed subjectively. There was no correlation between subjective and biochemical response. Biochemical progression (> or = 50% increase of nadir PSA) occurred after a median of 12 weeks. All but two patients were alive after a median follow-up time of 8.7 months for surviving patients (range 3.3-13.2). Eight patients experienced grade 3/4 leucopenia, with no indication of cumulative myelosuppression. Cardiovascular toxicity was experienced by four patients. Two patients developed angioedema twice, in one patient requiring hospitalization at the intensive ward. Based on this limited series, the combination of EPR and EMP in patients with HRPC is tolerable and appears to be effective in terms of significant PSA reduction. The results warrant further investigations of the two drugs and, in particular, of the clinical significance of > or = 50% PSA decrease in patients with HRPC. PMID- 9218740 TI - Basal cell carcinoma of the face: surgery or radiotherapy? Results of a randomized study. AB - Basal cell carcinomas (BCCs) are very frequent cutaneous cancers, often located on the face. Cure rates with surgery and radiotherapy are high, but these treatments have never been compared prospectively. A randomized trial was initiated in 1982 to compare surgery and radiotherapy in the treatment of primary BCC of the face measuring less than 4 cm. The primary end point was the failure rate (persistent or recurrent disease) after 4 years of follow-up. The secondary end point was the cosmetic results assessed by the patient, the dermatologist and three persons not involved in the trial. In the course of the trial, 347 patients were treated. Of the 174 patients in the surgery group, 71% had local anaesthesia and 91% frozen section examination. Of the 173 patients in the radiotherapy group, 55% were treated with interstitial brachytherapy, 33% with contactherapy and 12% with conventional radiotherapy. The 4-year actuarial failure rate (95% CI) was 0.7% (0.1-3.9%) in the surgery group compared with 7.5% (4.2-13.1%) in the radiotherapy group (log-rank P = 0.003). The cosmetic results assessed by four of the five judges were significantly better after surgery than after radiotherapy. Eighty-seven per cent of the surgery-treated patients and 69% of the radiation-treated patients considered the cosmetic result as good (P < 0.01). Thus, in the treatment of BCC of the face of less than 4 cm in diameter, surgery should be preferred to radiotherapy. PMID- 9218741 TI - Patients in phase I trials of anti-cancer agents in Japan: motivation, comprehension and expectations. AB - We attempted to characterize the motivation, comprehension and expectations of patients who had given informed consent to participate in phase I trials of anti cancer agents at the National Cancer Center of Japan. Thirty-three patients were given a simple multiple-choice questionnaire and asked to return it at a later date. The completed survey was returned by 32 patients. The patients were surveyed before they had received any investigational phase I agents. Nineteen per cent of patients were motivated to participate in the phase I trials by the possibility of therapeutic benefit, 9% because participation seemed a better choice than no treatment and only 6% for altruistic reasons. Most patients comprehended the major features of a phase I trial, namely its investigational nature, the unknown effects of the agent investigated and the unclear benefit to the patients themselves. Fifty-nine per cent of the patients anticipated that they might suffer severe or life-threatening side-effects if they participated in the phase I trial, and 43% were able to indicate accurately the purpose of the phase I trial as a dose determination study. Although only a minority of the patients indicated that their motivation to participate was possible treatment benefit to themselves, when answering questions regarding expectations, more than half indicated that there might be personal benefits of varying degrees by participation. PMID- 9218742 TI - Trends of Kaposi's sarcoma at AIDS diagnosis in Europe and the United States, 1987-94. AB - As a proportion of AIDS-defining illnesses, Kaposi's sarcoma (KS) decreased from 1987-89 to 1993-94 in homosexual and bisexual men in all European regions and in the United States. Albeit underestimated, AIDS KS rates in the general male population at ages 25-49 are higher than those of the majority of cancer sites in the same age group. PMID- 9218744 TI - Cutaneous malignant melanoma in Northern Ireland. AB - The results of two 5-year studies, for 1974-78 and 1984-88, of cutaneous malignant melanoma (CMM) in Northern Ireland show changes in the presentation of the disease. Although there is some evidence of earlier diagnosis, the rise in incidence has produced an overall increase in the number of cases with advanced disease. PMID- 9218743 TI - Breastfeeding history, pregnancy experience and risk of breast cancer. AB - Epidemiological evidence suggests that breastfeeding protects against breast cancer. Whether an effect of age at first breastfeeding is independent of an effect of age at first birth is unclear. We hypothesized that nausea and vomiting in pregnancy, which are associated with elevated serum oestradiol levels during pregnancy, may increase risk. Cases were 452 parous, premenopausal women, 40 years or younger, diagnosed with breast cancer in Los Angeles County from July 1983 to December 1988. Control subjects were matched to cases on age, race, parity and neighbourhood. Pregnancy and breastfeeding histories were obtained from in-person interviews. Odds of breast cancer among women who breastfed for at least 16 months relative to those among women who did not breastfeed was 0.66 [95% confidence interval (CI) 0.41-1.05]. Number of children breastfed was not associated with risk. Risk was lower in women who first breastfed at older ages. Having ever been treated for nausea or vomiting during pregnancy was associated with an increased risk, especially in women experiencing recent pregnancies (OR = 2.03, 95% CI 1.05-3.92). These results support a protective role of breastfeeding and an adverse role of nausea or vomiting during pregnancy in the development of premenopausal breast cancer, especially in the years immediately following pregnancy. PMID- 9218745 TI - Effects of a low-fat high-carbohydrate diet on plasma sex hormones in premenopausal women: results from a randomized controlled trial. Canadian Diet and Breast Cancer Prevention Study Group. AB - We are conducting a long-term randomized controlled trial to determine if intervention with a low-fat high-carbohydrate diet reduces breast cancer risk. The present study examines the effects of 2 years of dietary intervention on serum sex hormone levels in premenopausal women. Subjects with extensive mammographic densities were enrolled in a dietary intervention trial. The intervention involved intensive individual counselling aimed at reducing total dietary fat to 15% of calories. Control subjects received general advice about diet but were not counselled to change their fat intake. Serum sex hormone levels were measured in 220 premenopausal subjects at entry and 2 years after randomization. Two years after randomization oestradiol levels were 20% (70 pmol l(-1)) lower (P = 0.04) and progesterone levels were 35% (1.0 nmol l(-1)) lower (P = 0.004) and follicle-stimulating hormone (FSH) levels were 7% (1 IU) higher (P = 0.38) in the intervention group than in the control group. The FSH oestradiol ratio was 13% higher in the intervention group (P = 0.18). Samples analysed accounting for the timing of the blood sample in relation to the menstrual cycle showed that, in the intervention group, oestradiol and progesterone levels were lower and FSH levels higher in subjects with blood samples taken more than 30 days after the last menstrual period. Because of the strong evidence linking ovarian hormonal activity to breast cancer risk, these results suggest that a low-fat high-carbohydrate diet may reduce risk of breast cancer by reducing exposure to ovarian hormones that are a stimulus to cell division in the breast. PMID- 9218746 TI - Confocal microscopy of idarubicin localisation. PMID- 9218747 TI - Colorectal carcinoma: some reflections on bile flow through the terminal ileum. PMID- 9218748 TI - Pathological changes in the islet milieu precede infiltration of islets and destruction of beta-cells by autoreactive lymphocytes in a transgenic model of virus-induced IDDM. AB - RIP-LCMV transgenic mice that express the viral glycoprotein (GP) or nucleoprotein (NP) from lymphocytic choriomeningitis virus (LCMV) under control of the rat insulin promoter (RIP) in pancreatic beta-cells develop autoimmune diabetes (IDDM) after infection with LCMV. Previous reports have described that the viral infection activates naive, potentially autoreactive CD8+ cytotoxic T lymphocytes (CTL) that are present in the periphery of these mice, thus leading to the breaking of immunological unresponsiveness to the viral self-antigen expressed on beta-cells. However, we find that adoptive transfer of such CTL that were active in vitro and in vivo into uninfected RIP-LCMV recipients rarely resulted in hyperglycemia nor in insulitis, despite their ability to home to the islets and induce peri-insulitis. These observations indicated that, in addition to activated autoreactive lymphocytes, other factor(s) were required for beta cell destruction. The present study shows that upregulation of MHC class II molecules associated with the attraction/activation of antigen presenting cells (APCs) to the islets occurs as soon as 2 days after LCMV inoculation of transgenic mice, clearly before CD4+ and CD8+ lymphocytes are found entering the islets (days 6 and 7 after LCMV inoculation). In contrast, although some MHC class II upregulation is also found in islets of non-transgenic mice 2-4 days after LCMV infection, no insulitis or IDDM develops and MHC is downregulated to normal (pre-infection) levels by day 7-10 in these mice. Associated with the activation of APCs and MHC upregulation observed in transgenic mice, viral (LCMV) infection of islets was detectable 2 days post-viral inoculation in some mice. Thus, beta-cell destruction by activated autoreactive lymphocytes is a multifactorial process that is likely to require changes within the islet milieu or dysfunction of islets. PMID- 9218749 TI - Effect of beta-cell toxins on genetically engineered insulin-secreting cells. AB - The betacyte is a genetically engineered insulin-secreting liver cell line that is glucose responsive. Whether this cell is affected by specific beta-cell toxins is unknown. To explore this possibility we exposed these cells and those from the NIT-1 beta-cell line (positive controls) to the toxins streptozotocin (STZ, 2.5 20 mM), alloxan (ALL, 2.5-20 mM), and pentamidine (PENT, 10(-6)-1 mM). STZ and ALL were added for 1 h and pentamidine for 24 h. Insulin secretion from betacytes during a period of 5 h after removal of the toxin was inhibited only by pentamidine; all agents were inhibitory to NIT-1 cells. Glucose metabolism, as determined by a colorimetric MTT reduction assay, was adversely affected in betacytes by ALL (20 mM) and PENT (1 mM), and in NIT-1 cells by STZ (20 mM) as well as by ALL (2.5 mM) and PENT (1 mM). The magnitude of inhibition was less for the betacytes-58 v. 99%. Confluence of cells in culture wells and cell viability as assessed by the fluorochromes propidium iodide and acridine orange was reduced to a lesser extent for the betacytes than for the NIT-1 cells. The metabolic and microscopic effects of the toxins were unchanged in the betacyte from those in the liver cell line, HEP G2, from which the betacyte was engineered. These results of general resistance of the betacyte to beta-cell toxins with differing modes of action offer hope that this cell, or cells created in a similar manner from primary hepatocytes, may be at least partly resistant to the adverse effect of beta-cell toxins involved in autoimmune destruction of the pancreas. This increases the potential of the use of these cells for reversal of diabetes. PMID- 9218750 TI - Comparison of IA-2 with IA-2beta and with six other members of the protein tyrosine phosphatase family: recognition of antigenic determinants by IDDM sera. AB - To study the expression of protein tyrosine phosphatases (PTPs) in pancreatic islets, a cDNA library from islet cells was constructed and analysed. Twenty-one different PTPs were found to be expressed in islet cells, including three previously unknown PTPs. One of these, IA-2beta, was cloned, sequenced, and found to be related to IA-2, a major autoantigen in insulin-dependent diabetes mellitus (IDDM). The intracellular and extracellular domains of IA-2beta were 74 and 27% identical, respectively, to the intracellular and extracellular domains of IA-2. Approximately 70 and 45% of sera from patients with IDDM had autoantibodies that immunoprecipitated recombinant IA-2 and IA-2beta, respectively. By use of deletion mutants, we were able to show that the autoantibodies reacted with the intracellular, and not the extracellular, domains of IA-2 and IA-2beta, and that the major antigenic determinants resided within the COOH-terminus of the intracellular domains. Further studies revealed that approximately 97% of the IDDM sera that reacted with IA-2beta also reacted with IA-2, whereas only 50% of IDDM sera that reacted with IA-2 also reacted with IA-2beta. In contrast to the reactivity of IDDM sera with the IA-2 and IA-2beta, IDDM sera did not react with six other members of the PTP family. It is concluded that many members of the PTP family are expressed in pancreatic islets, but thus far only IA-2 and IA-2beta appear to be recognized as autoantigens by IDDM sera. PMID- 9218751 TI - Systemic production of interferon-gamma inducing factor (IGIF) versus local IFN gamma expression involved in the development of Th1 insulitis in NOD mice. AB - We report that the onset of Th1 insulitis is preceded by a rise of interferon gamma inducing factor (IGIF) mRNA expression in the spleen. This systemic shift towards Th1 reactivities was underlined by a close correlation of IGIF and IL 12p40 mRNA levels in the spleen, as determined by semi-quantitative RT-PCR. Cyclophosphamide-induced IGIF expression was also observed in MHC congenic NOR mice, but not in MHC class II-incompatible NON mice. The systemic rise of IGIF was followed by the development of destructive Th1-associated intra-insulitis. Interestingly, immunohistochemistry showed IL-4-positive cells evenly dispersed throughout the infiltrate, while IFN-gamma-positive cells were restricted to the vicinity of beta-cells. We conclude that cyclophosphamide induces a systemic shift in antigen presenting cells towards favouring Th1 responses, in an MHC dependent manner. Despite this general bias in immune reactivity, activation of Th1 cells in insulitis occurs only close to beta-cells, indicating a crucial role of antigen presentation by beta-cells or in their immediate vicinity. PMID- 9218752 TI - Development of insulitis and diabetes in B cell-deficient NOD mice. AB - Insulin-dependent diabetes mellitus (IDDM) is believed to be an autoimmune disease that results from autoimmune destruction of the insulin-secreting beta cells of the pancreas. In addition to a lymphocytic infiltration (insulitis) of the islets, patients with IDDM have autoantibodies directed against the components of the islet cells. Several beta-cell proteins have been identified as candidate autoantigens. The non-obese diabetic (NOD) mouse is a murine model for spontaneous IDDM. It is generally accepted that IDDM in patients and NOD mice results from the T lymphocyte-mediated destruction of beta-cells. However, the direct role of B lymphocytes in the disease process has not yet been clarified. To test directly the role of B cells in IDDM, we have generated B cell-deficient NOD mice by backcrossing the microMT-/- B cell 'knockout mice' onto the NOD background. The mice had no evidence of functional B cells as determined by flow cytometry and antibody production. We show that two out of seven of these mice developed insulitis and diabetes. These results suggest that despite an absence of B cells some NOD mice can still develop insulitis and diabetes. PMID- 9218753 TI - Diabetes results from a late change in the autoimmune response of NOD mice. AB - IDDM in the NOD mouse is the result of a chronic autoimmune process. NOD mice are shown to express benign autoimmunity that converts to a state of malignant autoimmunity and the development of IDDM. Young disease-prone NOD mice are in a state of benign autoimmunity that is correlated with a non-destructive response to islet tissue and the preservation of insulin-containing beta-cells. A proportion of mice with benign autoimmunity convert to having malignant autoimmunity. Clinical diabetes is diagnosed approximately 3 weeks from the development of malignant autoimmunity which is correlated with a destructive response to grafted islet tissue and extensive beta-cell destruction. We conclude that the development of clinical disease is correlated with a change in the state of autoimmunity, that is, from benign to malignant autoimmunity. PMID- 9218754 TI - BCG vaccination prevents insulin-dependent diabetes mellitus (IDDM) in NOD mice after disease acceleration with cyclophosphamide. AB - We have previously shown that immunotherapy with complete Freund's adjuvant (CFA) or BCG is highly effective in the prevention of spontaneous insulin-dependent diabetes mellitus (IDDM) and in circumventing the rejection of syngeneic islet grafts in diabetic NOD mice. This protection is reversed by treatment with cyclophosphamide (Cy). The present study was undertaken to determine the effect of BCG vaccination on the progression of Cy-accelerated diabetes in NOD mice and to understand the mechanism of BCG immunotherapy. The time course of Cy and BCG administration showed that the progression of Cy-induced diabetes can only be blocked when BCG vaccination is given within 3 days of Cy administration. Mice given BCG 3 days before (-3 days) or 7 days after Cy treatment were not protected. BCG immunization 1 day after Cy treatment almost completely prevented insulitis in the islets of Cy-treated mice. Cy treatment reduced the endogenous production of anti-GAD67 antibody, whereas BCG vaccination 1 day after Cy treatment restored the production of anti-GAD67 antibody of IgG1 isotype. The comprehensive effect of BCG vaccination on cytokine production in Cy-treated mice was to increase IL-4 production and change the IL-4/IFN-gamma ratio in both serum and supernatant of spleen cell cultures. We found that BCG-induced protection was associated with increased splenic CD4+CD45 RB(high) T cells. Taken together, our results indicate that BCG treatment counteracts the effect of Cy on autoimmune process in IDDM. However, BCG immunotherapy has a narrow window of up to 3 days after Cy treatment to block the progression of Cy-induced diabetes and to allow for the induction of regulatory cells which may effectively downregulate the diabetogenic cells. In summary, our results suggest that BCG vaccination prevents IDDM if given in the prediabetic state. After the induction of diabetes, disease progression can only be prevented within a narrow window of opportunity by this treatment. PMID- 9218755 TI - Flow cytometric study of T cell development in NOD mice reveals a deficiency in alphabetaTCR+CDR-CD8- thymocytes. AB - As a result of failed induction of T cell tolerance to pancreatic B cells, non obese diabetic (NOD) mice develop spontaneous autoimmune insulin-dependent diabetes mellitus (IDDM). The thymic stroma, which plays a crucial role in thymic T cell maturation, undergoes extensive premature disorganization in NOD mice, so it is of interest to examine NOD T cell development. In this study, both major and minor developmental populations of thymocytes of NOD/Lt mice were studied and compared to those of BALB/c, C57BL/6 and CBA mice by multiparameter flow cytometry (FACS). These results are described in detail and reveal that most thymocyte subsets were normally represented, including alphaTcR-CD4-CD8- (triple negative; TN), alphabetaTcR-CD4+CD8- and alphabetaTcR-CD4-CD8+ (immature single positive; ISP), alphabetaTcR-/lowCD4+CD8+ (double positive; DP) and alphabetaTcR+CD4+CD8- and alphabetaTcR+CD4-CD8+ (mature single positive; SP) as well as gammadelta T cells. However, NOD mice exhibited a marked deficiency of thymic alphabetaTcR+CD4-CD8- (alphabeta+DN) T cells. alphabeta+DN T cells, which are included among NK1+ T cells in C57BL/6 mice, produce large amounts of IL-4 on primary stimulation. Given the potential significance of NKT cells in immunoregulation, it is possible that the scarcity of these cells in NOD mice plays a role in the pathogenesis of IDDM. PMID- 9218756 TI - Immunization therapies in the prevention of diabetes. AB - Insulin-dependent diabetes (IDD), being an autoimmune disease, offers several opportunities for immunological interventions that may result either in the reduction of disease severity or in delaying diabetes onset. Among the various experimental preventative approaches, parenteral immunization with islet-specific autoantigens appears to be practically simpler and promising. We have previously shown that immunization with insulin, insulin B chain and B chain epitope (p9 23), but not insulin A chain, in incomplete Freund's adjuvant (IFA) and in alum (with B chain) delayed/prevented diabetes onset in NOD mice. Here we demonstrate the protective efficacy of affinity purified GAD65 in IFA. While both insulin B chain and GAD65 significantly delayed the onset of diabetes (P=0.001), a recently described tyrosine phosphatase (IA-2) antigen did not (P=0.38). Interestingly, B chain immunization reduced the incidence of cyclophosphamide (CY)-accelerated diabetes by about 50-55%. We also provide further evidence that B chain, upon increased adsorption to alum, could improve on its protective capacity in NOD mice. PMID- 9218758 TI - Molecular role of TGF-beta, secreted from a new type of CD4+ suppressor T cell, NY4.2, in the prevention of autoimmune IDDM in NOD mice. AB - A new type of CD4+ T cell clone (NY4.2) isolated from pancreatic islet infiltrated lymphocytes of acutely diabetic non-obese diabetic (NOD) mice prevents the development of insulin-dependent diabetes mellitus (IDDM) in NOD mice, as well as the recurrence of autoimmune diabetes in syngeneic islet transplanted NOD mice. It has been demonstrated that the cytokine TGF-beta, secreted from the cells of this clone, is the substance which prevents autoimmune IDDM. This investigation was initiated to determine the molecular role TGF-beta plays in the prevention of autoimmune IDDM by determining its effect on IL-2 induced signal transduction in Con A-activated NOD mouse splenocytes and HT-2 cells. First, we determined whether TGF-beta, secreted from NY4.2 T cells, inhibits IL-2-dependent T cell proliferation in HT-2 cells (IL-2-dependent T cell line) and NOD splenocytes. We found that TGF-beta suppresses IL-2-dependent T cell proliferation. Second, we determined whether TGF-beta inhibits the activation of Janus kinases (JAKs), as well as signal transducers and activators of transcription (STAT) proteins, involved in an IL-2-induced signalling pathway that normally leads to the proliferation of T cells. We found that TGF-beta inhibited tyrosine phosphorylation of JAK1, JAK3, STAT3 and STAT5 in Con A blasts from NOD splenocytes and HT-2 cells. Third, we examined whether TGF-beta inhibits the cooperation between STAT proteins and mitogen-activated protein kinase (MAPK), especially extracellular signal-regulated kinase 2 (ERK2). We found that TGF-beta inhibited the association of STAT3 and STAT5 with ERK2 in Con A blasts from NOD splenocytes and HT-2 cells. On the basis of these observations, we conclude that TGF-beta may interfere with signal transduction via inhibition of the IL-2-induced JAK/STAT pathway and inhibition of the association of STAT proteins with ERK2 in T cells from NOD splenocytes, resulting in the inhibition of IL-2-dependent T cell proliferation. TGF-beta-mediated suppression of T cell activation may be responsible for the prevention of effector T cell-mediated autoimmune IDDM in NOD mice by TGF-beta-producing CD4+ suppressor T cells. PMID- 9218757 TI - Protection of NIT-1 pancreatic beta-cells from immune attack by inhibition of NF kappaB. AB - We have recently observed that inhibition of NF-kappaB in NIT-1 insulinoma cells protects them from tumour necrosis factor (TNF)-induced cell death in vitro, possibly because expression of interleukin-1 (IL-1)beta-converting enzyme (ICE), a member of the cysteine protease pathway of cell death, is decreased. In the current study we have examined the effect of the same inhibitor of NF-kappaB on class I major histocompatibility complex (MHC) protein expression in NIT-1 cells and shown that inhibition of NF-kappaB activation decreased basal and TNF-induced class I MHC levels. Although inducible nitric oxide synthase (iNOS) may also be inhibited by inhibition of NF-kappaB, this could not be demonstrated in NIT 1/delta sp cells because wild-type NIT-1 cells express very little iNOS. When NIT 1/delta sp12 cells, expressing high levels of the NF-kappaB inhibitor, are transplanted into immunodeficient NOD/scid mice, tumorigenesis and death by hypoglycemia proceed similarly to untransfected NIT-1 cells. Untransfected NIT-1 cells were killed by co-transfer of splenic T cells from diabetic but not non diabetic NOD mice. NIT-1/delta sp12 cells were protected from killing in vivo by T cells from diabetic mice, in that tumours developed in four out of five mice and the kinetics of tumour development were not significantly delayed. NIT 1/delta sp12 cells were not protected from killing by T cells from mice previously primed with NIT-1 cells. In conclusion, inhibition of NF-kappaB is likely to suppress several different pathways of immune-mediated cell death in beta-cells and protects NIT-1 cells from immune attack by diabetogenic T cells in vivo. Inhibition of NF-kappaB is a potentially effective strategy for protection of pancreatic beta-cells in autoimmune diabetes. PMID- 9218759 TI - Rat pancreatic islet and skin xenograft survival in CD4 and CD8 knockout mice. AB - The relative contributions of the CD4+ and CD8+ T cell subpopulations in xenotransplant rejection were studied using CD4 and CD8 knockout (KO) mice. Wistar Furth (WF, RT1a) rat pancreatic islet or skin xenografts were transplanted into either CD4 or CD8 KO recipients and compared to wild-type controls. Long term survival of WF islet xenografts was observed in the CD4 KO mice (MST, >66+/ 8 days) whereas CD8 KO mice rejected their islet xenografts within 8 days, similar to controls (MST, 7+/-0.2 days). In contrast, WF skin xenografts were rejected in both CD4 and CD8 KO recipients within 8 days. CD4 KO recipients which maintained xenoislets >90 days posttransplant rejected WF skin grafts within 9 days, without rejecting their original islet xenografts. These results suggest that CD4+ cells are essential for mediating islet xenograft rejection. These data also suggest that the absence of either CD4+ or CD8+ T cells is not sufficient to prevent rejection of skin xenografts. PMID- 9218760 TI - T cell receptor gene polymorphisms associated with anti-insulin, autoimmune T cells in diabetes-prone NOD mice. AB - The great majority of insulin-specific T cell clones isolated from islets of NOD mice react with insulin peptide B-(9-23) (amino acids 9-23 of the insulin B chain). The T cell receptors of these clones contain diverse beta-chains but restricted alpha-chains. The dominant alpha-chain motif is a V alpha 13 segment (10 out of 13) combined to either a J alpha 45 or a J alpha 34 segment (eight out of 13). Furthermore, nine out of 10 of these V alpha 13 segments are a product of a novel NOD TCR V alpha gene which we have termed V alpha 13.3. Analysis of V alpha 13 transcripts from splenic cDNA libraries from the NOD, BALB/c and C57BL/6 mice revealed significant differences between strains. The NOD sequences contained both V alpha 13.1 and the novel V alpha 13.3. The BALB/c contained the previously reported V alpha 13.1 and V alpha 13.2, but not the V alpha 13.3 sequence identified in the NOD anti-insulin T cell clones. The C57BL/6 had V alpha 13.1 and V alpha 13.3 plus two additional novel sequences which we have termed V alpha 13.4 and V alpha 13.5. These V alpha 13 subfamily members differed by two to four amino acids in either the CDR1 region or adjoining the CDR2 region. The frequency of utilization of the different V alpha 13 subtypes varied dramatically between strains. In the NOD spleen, V alpha 13.3 was detected 79% of the time, compared to 21% for V alpha 13.1. In contrast, the C57BL/6 spleen contained only 7% of V alpha 13.3 sequences compared to the other subfamily members present (V alpha 13.1: 27%; V alpha 13.4: 56%; V alpha 13.5: 10%). MHC polymorphisms or other unknown selective pressures may contribute to these differences in V alpha 13 utilization. We hypothesize that the presence and frequent utilization of the V alpha 13.3 T cell receptor element is involved in targeting insulin B-(9-23) and may be related to diabetes susceptibility of NOD mice. PMID- 9218761 TI - Roles for motility in bacterial-host interactions. AB - The ability to move in a directed manner may confer distinct advantages upon host adapted prokaryotes. Potential benefits of motility include increased efficiency of nutrient acquisition, avoidance of toxic substances, the ability to translocate to preferred hosts and access optimal colonization sites within them, and dispersal in the environment during the course of transmission. The costs of motility also may be significant. These include the metabolic burden of synthesizing flagellar components, the energetic expense of fuelling flagellar motors and the presentation of polymeric and highly antigenic targets to the immune system. It is therefore not surprising that synthesis of the motility apparatus is usually subject to strict control. Studies of a variety of bacterial host interactions demonstrate roles for motility, and its regulation, at points throughout the infectious cycle. PMID- 9218762 TI - Sulphation of Rhizobium sp. NGR234 Nod factors is dependent on noeE, a new host specificity gene. AB - Rhizobia secrete specific lipo-chitooligosaccharide signals (LCOs) called Nod factors that are required for infection and nodulation of legumes. In Rhizobium sp. NGR234, the reducing N-acetyl-D-glucosamine of LCOs is substituted at C6 with 2-O-methyl-L-fucose which can be acetylated or sulphated. We identified a flavonoid-inducible locus on the symbiotic plasmid pNGR234a that contains a new nodulation gene, noeE, which is required for the sulphation of NGR234 Nod factors (NodNGR). noeE was identified by conjugation into the closely related Rhizobium fredii strain USDA257, which produces fucosylated but non-sulphated Nod factors (NodUSDA). R. fredii transconjugants producing sulphated LCOs acquire the capacity to nodulate Calopogonium caeruleum. Furthermore, mutation of noeE (NGRdelta noeE) abolishes the production of sulphated LCOs and prevents nodulation of Pachyrhizus tuberosus. The sulphotransferase activity linked to NoeE is specific for fucose. In contrast, the sulphotransferase NodH of Rhizobium meliloti seems to be less specific than NoeE, because its introduction into NGRdelta noeE leads to the production of a mixture of LCOs that are sulphated on C6 of the reducing terminus and sulphated on the 2-O-methylfucose residue. Together, these findings show that noeE is a host-specificity gene which probably encodes a fucose-specific sulphotransferase. PMID- 9218763 TI - mRNA stability is regulated by a coding-region element and the unique 5' untranslated leader sequences of the three Synechococcus psbA transcripts. AB - The psbAI and psbAIII transcripts in Synechococcus sp. strain PCC 7942 are subject to accelerated turnover when cells are exposed to high light intensities, but psbAII message stability is unaffected. We used a psbAI 'minigene' which has a part of the coding sequence removed as a reporter gene in order to identify the cis-acting elements of the transcript that determine stability. While engineering the minigene to optimally mimic the native gene, we identified a stabilizer element within the open reading frame, corresponding to the coding region for the first membrane span of the D1 protein, the presence of and translation through which was essential for normal psbA mRNA stability. We propose that this stabilizer is a site for ribosome pausing, and that accumulation of ribosomes on the transcript upstream of the pause site increases stability. To identify the elements that regulate the differential responses of the psbA transcripts to high light growth, sequences from psbAII and psbAIII were substituted in the psbAI minigene reporter. The chimeric reporter transcripts established that the psbAI and psbAIII untranslated leaders determine the faster turnover of these messages. The untranslated leader regions of the psbA transcripts may regulate mRNA stability by modulating translation and thereby stability, or by recruiting RNA binding proteins that affect mRNA turnover more directly. PMID- 9218764 TI - DNA binding of Escherichia coli arginine repressor mutants altered in oligomeric state. AB - The Escherichia coli arginine repressor (ArgR) controls expression of the arginine biosynthetic genes and acts as an accessory protein in Xer site-specific recombination at cer and related plasmid recombination sites. The hexameric wild type protein shows L-arginine-dependent DNA binding. In this work, ArgR mutants that are defective in trimer-trimer interactions and bind DNA as trimers in an L arginine-independent manner are isolated and characterized. Whereas the wild-type ArgR hexamer exhibits high-affinity binding to two repeated ARG boxes separated by 3 bp (each ARG box containing two identical dyad symmetrical 9 bp half-sites), the trimeric mutants bind to and footprint three adjacent half-sites of this 'idealized' substrate. Trimeric ArgR is impaired in its ability to repress the arginine biosynthetic genes and in Xer site-specific recombination. In the absence of L-arginine, residual wild-type ArgR-binding occurs as trimers. The binding of an N-terminal 77-amino-acid DNA-binding domain to idealized ARG boxes is also characterized. PMID- 9218765 TI - Genes involved in conjugative DNA processing of plasmid R6K. AB - The conjugative transfer region of the IncX plasmid R6K (TRA(x)) was analysed by transposon mutagenesis and DNA sequencing. Tn5tac1 insertional mutations localized TRA(x) to a 14.8 kb segment containing the alpha origin of transfer (oriT alpha), genes involved in conjugative DNA-processing (Dtr(x)) and genes involved in pilus synthesis and assembly (Mpf(x)). A second functional oriT, oriTbeta, was located at a distance of 5.3 kb from oriT alpha and was outside TRA(x). Mpf(x) occupied a segment of 10kb, as judged by the location of insertions conferring resistance to infection by the X pilus-specific phage X-2. At both sides of Mpf(x) there were insertions that were Tra but X-2 sensitive, suggesting that the mutations were in Dtr(x). This region was sequenced and three genes were identified: taxA, taxB, and taxC. The overall organization was oriT alpha-taxA-taxC-Mpf(x)-taxB. taxC coded for a oriT-relaxase that belongs to the VirD2 family. taxA coded for a protein of 181 amino acids that showed similarity to TraY of F-like plasmids and to the Arc-repressor superfamily. TaxB showed similarity to TraG-like proteins, a protein superfamily probably involved in coupling the relaxosome to the DNA-transport apparatus. TaxA and TaxC are required for oriT nicking in vivo. The nicking reaction was mistakenly assumed by Flashner et al. (1996) to represent a feature of the vegetative replication origins. However, insertions or deletions disrupting taxA and taxC affected conjugation but not replication of R6K. Conversely, protein pi, which is absolutely required for replication of R6K, was not required for conjugative transfer. In addition, protein DDP3, which is also assumed to have a role in replication, was found to be a positive modulator of bacterial conjugation. Taken together, these results rule out a direct and essential involvement of conjugation proteins in R6K vegetative replication, and also rule out the requirement of replication protein pi for conjugation. PMID- 9218766 TI - Regulation of the xcp secretion pathway by multiple quorum-sensing modulons in Pseudomonas aeruginosa. AB - The virulence of the opportunistic pathogen Pseudomonas aeruginosa is largely dependent upon the extracellular production of a number of secreted proteins with toxic or degradative activities. The synthesis of several exoenzymes is controlled in a cell-density-dependent manner by two interlinked quorum-sensing systems. Their secretion across the outer membrane occurs through the Xcp translocation machinery. The xcp locus located at 40 min on the chromosome consists of two divergently transcribed operons, namely xcpPQ and xcpR to xcpZ. In this study, transcriptional fusions were constructed between the xcpP and xcpR genes and the lacZ reporter. Transcriptional activation of the xcpP and xcpR genes in P. aeruginosa is growth-phase dependent and the lasR-lasI autoinduction system is required for this control. In the heterologous host Escherichia coli, the lasR gene product, together with its cognate autoinducer N-(3-oxododecanoyl) L-homoserine lactone (OdDHL), activates both the xcpP-lacZ and the xcpR-lacZ gene fusion. The second P. aeruginosa quorum-sensing modulon rhIR-rhII (vsmR-vsmI) is also involved in the control of the xcp genes. Expression of the lacZ fusions is strongly reduced in PANO67, a pleiotropic mutant defective in the production of N acyl-homoserine lactones responsible for the activation of RhIR. Furthermore, introduction of the lasR mutation in PANO67 results in additional diminution of xcpR transcription, indicating that the two systems can regulate their target genes independently. These data demonstrate that expression of the xcp secretion system depends on a complex regulatory network involving cell-cell signalling which controls production and secretion of virulence-associated factors. PMID- 9218767 TI - Adherence of Porphyromonas gingivalis to matrix proteins via a fimbrial cryptic receptor exposed by its own arginine-specific protease. AB - Porphyromonas gingivalis, a Gram-negative anaerobe, is known to be involved in the pathogenesis of periodontitis. P. gingivalis fimbriae, which are proteinaceous appendages extending from the cell surface, may contribute to the adherence of the organism to the host cell surface. We previously suggested that arginine-specific protease produced by P. gingivalis enhanced the adherence of purified fimbriae to fibroblasts or matrix proteins. In this study, we have revealed the mechanism of the enhanced binding of fimbriae by the protease in more detail. Arg-specific protease and fimbriae were obtained from P. gingivalis 381 cells and purified. We then analysed the interaction of fimbriae and immobilized fibronectins (intact or partially degraded fibronectin by the purified protease) by using the real-time biomolecular interaction analysis (BIAcore) system with an optical biosensor based on the principles of surface plasmon resonance. BIAcore profiles demonstrated an enhanced interaction between fimbriae and protease-degraded fibronectin. We also showed specific binding of fimbriae to the degraded fibronectin by means of BIAcore analysis. The binding of biotinylated fimbriae to immobilized fibronectin was examined by enzyme-linked biotin-avidin assay. The purified protease enhanced the fimbrial binding to the immobilized fibronectin. The enhancement was inhibited by the addition of L-Arg, or oligopeptides containing the Arg residue at the C-terminus in the fimbrial binding reaction, suggesting that the P. gingivalis fimbriae may potentially have an ability to bind tightly to the Arg residue at C-terminus. Taken together, these studies indicate that P. gingivalis arginine-specific protease can expose a cryptitope in the matrix protein molecules, i.e. the C-terminal Arg residue of the host matrix proteins, so that the organism can adhere to the surface layer in the oral cavity through fimbriae-Arg interaction (a novel host-parasite relationship). PMID- 9218768 TI - Mutations in a dispensable region of the UaY transcription factor of Aspergillus nidulans differentially affect the expression of structural genes. AB - The uaY gene encodes a transcriptional activator mediating uric acid induction of at least nine genes of the purine-utilization pathway. In this article, we characterize a loss-of-function mutation, uaY205, as a 16 bp deletion that results in premature translation termination, and substitutes the C-terminal 63 amino acids for 13 amino acid residues. Reversion analysis demonstrates that the C-terminal 63 amino acid residues are unnecessary for UaY function, and that the loss-of-function phenotype resulting from the uaY205 mutation is caused by the new amino acid sequence present in the mutant protein. Revertants in two different frames (wild type and +1) restore function but show subtle differences in the expression of genes controlled by the UaY protein. Two strains showing elevated expression of genes under UaY control were shown to carry, in addition to a mutation leading to the recovery of the wild-type open reading frame, mutations in unlinked genes. Using crude extracts of Aspergillus nidulans, we have been able to detect, for the first time, in transcription factors of this class, specific retardation of a promoter probe. The binding activity is at least partially dependent on the presence of inducer. The gel shift experiments show that the novel inhibitory sequence present in the UaY205 protein can act either by affecting the stability of the protein, or via an inter- or intramolecular interaction impairing the specific DNA-binding activity. PMID- 9218769 TI - Cloning and analysis of a Borrelia burgdorferi membrane-interactive protein exhibiting haemolytic activity. AB - We cloned the gene encoding a membrane-interactive protein of Borrelia burgdorferi by means of its haemolytic activity in Escherichia coli. The haemolytic activity was erythrocyte-species specific, with progressively decreasing activity for erythrocytes from horse, sheep, and rabbit, respectively. Genetic analysis of the haemolytic determinant revealed two borrelia haemolysin genes, blyA and blyB, that are part of a predicted four-gene operon which is present in multiple copies on the 30 kb circular plasmid(s) of B. burgdorferi B31. blyA encodes a predicted alpha-helical 7.4 kDa protein with a hydrophobic central region and a positively charged C-terminus, which is structurally homologous to a large group of pore-forming toxins with cytolytic activity. blyB encodes a soluble protein which stabilized BlyA and enhanced haemolytic activity. While the majority of BlyA in E. coli was membrane-associated, only soluble protein was haemolytically active. The haemolytic activity was shown to be highly protease sensitive, heat labile, independent of divalent cations, and extremely dependent on protein concentration, consistent with a requirement for oligomerization as the mechanism of action. BlyA was highly purified from E. coli in a single step utilizing Triton X-114 phase partitioning. Genetic analysis of blyA and blyB mutants indicated that the stability, membrane association, and activity of BlyA was dependent on subtle changes in its sequence and on the BlyB protein. The bly genes were found to be expressed at a very low level in cultured B. burgdorferi. PMID- 9218770 TI - Genetic analysis of pertussis toxin promoter activation in Bordetella pertussis. AB - Bordetella pertussis regulates expression of its virulence factors such as pertussis toxin (Ptx) via the bvg locus, which encodes a two-component system composed of a sensor protein, BvgS, and a transcription activator, BvgA. We used a ptx-lac fusion on the B. pertussis chromosome to analyse promoter activation by alteration of specific sequences upstream of and within the promoter. Our data demonstrate that a pair of heptanucleotide inverted repeats separated by a turn of the DNA helix within the upstream repeat region (centred around nucleotide 136.5) are crucial cis-activating elements, and probably represent the initial BvgA-binding site. In addition, we demonstrate that the sequence between these repeats and the promoter plays a role in activation. Our data are most consistent with a model of co-operative binding of BvgA dimers to this intervening region and interaction with RNA polymerase at the promoter to activate ptx transcription. In the core promoter region both the non-consensus 21 bp spacing and the specific sequence between the -35 and -10 elements are crucial for promoter activity. PMID- 9218771 TI - Isolation of a dnaE mutation which enhances RecA-independent homologous recombination in the Escherichia coli chromosome. AB - The mechanism of recombination of tandem repeats in the chromosome of Escherichia coli was investigated by genetic means. Tandem repeats 624 bp long were introduced into the lacZ gene of E. coli and the efficiency of deletion of one repeat was compared in different recombination mutants. No effects of the recA, recBC, recF, ruvA or ruvA recG mutations were detected. Hence, tandem repeat deletion appears to not proceed via the RecBCD or RecF homologous recombination pathways. A new mutant in which RecA-independent recombination is increased 15 fold was isolated. The mutation lies in the dnaE gene coding for the alpha subunit of polymerase III: it is a Gly to Asp change at codon 133. Another dnaE mutation, dnaE486, was tested and also shown to stimulate RecA-independent recombination. It is proposed that tandem-repeat recombination occurs by a replication slippage mechanism. RecA-independent recombination is also enhanced in a rep mutant, in which chromosomal replication is slowed down by the absence of the Rep helicase, suggesting that replication pausing may facilitate slippage. PMID- 9218772 TI - Role of arginine-43 and arginine-69 of the Hin recombinase catalytic domain in the binding of Hin to the hix DNA recombination sites. AB - The Hin recombinase mediates the site-specific inversion of a segment of the Salmonella chromosome between two flanking 26bp hix DNA recombination sites. Mutations in two amino acid residues, R43 and R69 of the catalytic domain of the Hin recombinase, were identified that can compensate for loss of binding resulting from elimination of certain major and minor groove contacts within the hix recombination sites. With one exception, the R43 and R69 mutants were also able to bind a hix sequence with an additional 4bp added to the centre of the site, unlike wild-type Hin. Purified Hin mutants R43H and R69C had both partial cleavage and inversion activities in vitro while mutants R43L, R43C, R69S, and R69P had no detectable cleavage and inversion activities. These data support a model in which the catalytic domain plays a role in DNA-binding specificity, and suggest that the arginine residues at positions 43 and 69 function to position the Hin recombinase on the DNA for a step in the recombination reaction which occurs either at and/or prior to DNA cleavage. PMID- 9218773 TI - Identification of Pseudomonas aeruginosa genes required for epithelial cell injury. AB - We have developed a simple, reproducible and rapid genetic screen for Pseudomonas aeruginosa-induced epithelial cell cytotoxicity in cultures of MDCK cells. This screen was used to isolate isogenic transposon-tagged non-cytotoxic mutants of a cytotoxic and lung-virulent strain of P. aeruginosa (PA103). The transposon insertion site was determined by using an inverse polymerase chain reaction followed by DNA-sequence analysis. On the basis of phenotype and sequence analysis, these mutants fell into four classes. One class had absent or defective pill, based on their resistance to phage PO4 and/or loss of twitching motility (twt-). A second class exhibited decreased adherence. A third class of mutants exhibited probable defects in the machinery or targets of type III protein secretion. A final class of mutants exhibited decreased but not absent cytotoxicity. This class included members of the first three classes as well as other mutants. These results suggest that localized cytotoxicity is likely to require several steps and several components, including pili and other (unidentified) extracellular proteins. The type III protein-secretion apparatus appears to be involved in this process. PMID- 9218774 TI - ftsW is an essential cell-division gene in Escherichia coli. AB - In the absence of exogenous promoters, plasmid-mediated complementation of the temperature-sensitive ftsW201 allele requires the presence of the full coding sequence of ftsW plus upstream DNA encompassing the C-terminus of mraY and the full coding sequence of murD. We used molecular and genetic techniques to introduce an insertional inactivation into the chromosomal copy of ftsW, in the presence of the plasmid-borne wild-type ftsW gene under the control of P(BAD). In the absence of arabinose, the ftsW-null strain is not viable, and a shift from arabinose- to glucose-containing liquid medium resulted in a block in division, followed by cell lysis. Immunofluorescence microscopy revealed that in ftsW-null filaments, the FtsZ ring is absent in 50-60% of filaments, whilst between one and three Z-rings per filament can be detected in the remainder of the population, with the majority of these containing only one Z-ring per filament. We also demonstrated that the expression of only ftsWS (the smaller of two ftsW open reading frames) from P(BAD) is sufficient for complementation of the ftsW-null allele. We conclude that FtsW is an essential cell-division protein in Escherichia coli, and that it plays a role in the stabilization of the FtsZ ring during cell division. PMID- 9218775 TI - Cloning, sequence and mutagenesis of the structural gene of Pseudomonas aeruginosa CysB, which can activate algD transcription. AB - Pseudomonas aeruginosa strains infecting patients with cystic fibrosis (CF) acquire a mucoid phenotype due to overproduction of alginate. The key enzyme in alginate synthesis is AlgD, whose promoter is transcriptionally active in mucoid strains and under the control of several trans-acting factors, including the integration host factor (IHF). The algD promoter (palgD) contains two IHF-binding sites (ihf1 and ihf2). Study of IHF binding to ihf2 of palgD, by electrophoretic mobility-shift assays, led to the discovery of a protein of 36 kDa (p36) able to bind downstream from ihf2, to the 3' region of palgD. The gene encoding p36 was isolated from the mucoid strain CHA of P. aeruginosa and sequenced. It can encode a 324-amino-acid protein, which shares a high degree of sequence identity (63%) with CysB from Escherichia coli and from Salmonella typhimurium, a transcriptional factor of the LysR superfamily. Furthermore, both p36 and S. typhimurium CysB bind the same site of palgD; p36 was therefore termed CysB and its structural gene was called cysB. Next to cysB, on the opposite DNA strand, cysH was capable of encoding a protein sharing 26% identity with CysH (PAPS reductase) of E. coli and an even greater identity (54%) with the nucleotide deduced protein from Arabidopsis. A CysB-deficient mutant of CHA, constructed by insertional inactivation of cysB, was a cysteine auxotroph and was unable to form a specific complex with palgD in vitro. Activity of palgD in the cysB mutant, in CHA and in the non-mucoid strain PAO was assessed by the use of a transcriptional algD-xylE fusion. Cells of PAO and of the cysB mutant grown in minimal media in the presence of 0.3 M NaCl exhibited a palgD activity, which was 10% or less that of the mucoid strain CHA. Thus, P. aeruginosa CysB can act as an activator of algD expression. PMID- 9218776 TI - Identification of a bacterial pectin acetyl esterase in Erwinia chrysanthemi 3937. AB - Erwinia chrysanthemi causes soft-rot diseases of various plants by enzymatic degradation of the pectin in plant cell walls. The structural complexity of pectin requires the combined action of several pectinases for its efficient breakdown. Three types of pectinases have so far been identified in E. chrysanthemi: two pectin methyl esterases (PemA, PemB), a polygalacturonase (PehX), and eight pectate lyases (PelA, PelB, PelC, PelD, PelE, PelL, PelZ, PelX). We report in this paper the analysis of a novel enzyme, the pectin acetyl esterase encoded by the paeY gene. No bacterial form of pectin acetyl esterases has been described previously, while plant tissues and some pectinolytic fungi were found to produce similar enzymes. The paeY gene is present in a cluster of five pectinase-encoding genes, pelA-pelE-pelD-paeY-pemA. The paeY open reading frame is 1650 bases long and encodes a 551-residue precursor protein of 60704Da, including a 25-amino-acid signal peptide. PaeY shares one region of homology with a rhamnogalacturonan acetyl esterase of Aspergillus aculeatus. To characterize the enzyme, the paeY gene was overexpressed and its protein product was purified. PaeY releases acetate from sugar-beet pectin and from various synthetic substrates. Moreover, the enzyme was shown to act in synergy with other pectinases. The de-esterification rate by PaeY increased after previous demethylation of the pectins by PemA and after depolymerization of the pectin by pectate lyases. In addition, the degradation of sugar-beet pectin by pectate lyases is favoured after the removal of methyl and acetyl groups by PemA and PaeY, respectively. The paeY gene was first identified on the basis of its regulation, which shares several characteristics with that of other pectinases. Analysis of the paeY transcription, using gene fusions, revealed that it is induced by pectic catabolic products and is affected by growth phase, oxygen limitation and catabolite repression. Regulation of paeY expression appears to be dependent on the KdgR repressor, which controls all the steps of pectin catabolism, and on the catabolite regulatory protein (CRP), the global activator of sugar catabolism. The contiguous pelD, paeY and pemA genes are transcribed as an operon from a promoter proximal to pelD which allows the regulation by KdgR and CRP. However, transcription can be interrupted at the intra-operon Rho independent terminator situated between pelD and paeY. The paeY mutant inoculated into Saintpaulia plants was less invasive than the wild-type E. chrysanthemi strain 3937, demonstrating the important role of PaeY in the soft-rot disease. PMID- 9218778 TI - Sequence interruptions in enterobacterial repeated elements retain their ability to encode well-folded RNA secondary structures. PMID- 9218777 TI - Proteolysis of the phage lambda CII regulatory protein by FtsH (HflB) of Escherichia coli. AB - Rapid proteolysis plays an important role in regulation of gene expression. Proteolysis of the phage lambda CII transcriptional activator plays a key role in the lysis-lysogeny decision by phage lambda. Here we demonstrate that the E. coli ATP-dependent protease FtsH, the product of the host ftsH/hflB gene, is responsible for the rapid proteolysis of the CII protein. FtsH was found previously to degrade the heat-shock transcription factor sigma32. Proteolysis of sigma32 requires, in vivo, the presence of the DnaK-DnaJ-GrpE chaperone machine. Neither DnaK-DnaJ-GrpE nor GroEL-GroES chaperone machines are required for proteolysis of CII in vivo. Purified FtsH carries out specific ATP-dependent proteolysis of CII in vitro. The degradation of CII is at least 10-fold faster than that of sigma32. Electron microscopy revealed that purified FtsH forms ring shaped structures with a diameter of 6-7 nm. PMID- 9218779 TI - Insertions within ERIC sequences. PMID- 9218780 TI - Crystal structure and mechanism of human L-arginine:glycine amidinotransferase: a mitochondrial enzyme involved in creatine biosynthesis. AB - L-arginine:glycine amidinotransferase (AT) catalyses the committed step in creatine biosynthesis by formation of guanidinoacetic acid, the immediate precursor of creatine. We have determined the crystal structure of the recombinant human enzyme by multiple isomorphous replacement at 1.9 A resolution. A telluromethionine derivative was used in sequence assignment. The structure of AT reveals a new fold with 5-fold pseudosymmetry of circularly arranged betabeta alphabeta-modules. These enclose the active site compartment, which is accessible only through a narrow channel. The overall structure resembles a basket with handles that are formed from insertions into the betabeta alphabeta-modules. Binding of L-ornithine, a product inhibitor, reveals a marked induced-fit mechanism, with a loop at the active site entrance changing its conformation accompanied by a shift of an alpha-helix by -4 A. Binding of the arginine educt to the inactive mutant C407A shows a similar mode of binding. A reaction mechanism with a catalytic triad Cys-His-Asp is proposed on the basis of substrate and product bound states. PMID- 9218781 TI - Crystal structure of human glyoxalase I--evidence for gene duplication and 3D domain swapping. AB - The zinc metalloenzyme glyoxalase I catalyses the glutathione-dependent inactivation of toxic methylglyoxal. The structure of the dimeric human enzyme in complex with S-benzyl-glutathione has been determined by multiple isomorphous replacement (MIR) and refined at 2.2 A resolution. Each monomer consists of two domains. Despite only low sequence homology between them, these domains are structurally equivalent and appear to have arisen by a gene duplication. On the other hand, there is no structural homology to the 'glutathione binding domain' found in other glutathione-linked proteins. 3D domain swapping of the N- and C terminal domains has resulted in the active site being situated in the dimer interface, with the inhibitor and essential zinc ion interacting with side chains from both subunits. Two structurally equivalent residues from each domain contribute to a square pyramidal coordination of the zinc ion, rarely seen in zinc enzymes. Comparison of glyoxalase I with other known structures shows the enzyme to belong to a new structural family which includes the Fe2+-dependent dihydroxybiphenyl dioxygenase and the bleomycin resistance protein. This structural family appears to allow members to form with or without domain swapping. PMID- 9218782 TI - Structure of the PH domain and Btk motif from Bruton's tyrosine kinase: molecular explanations for X-linked agammaglobulinaemia. AB - Bruton's tyrosine kinase (Btk) is an enzyme which is involved in maturation of B cells. It is a target for mutations causing X-linked agammaglobulinaemia (XLA) in man. We have determined the structure of the N-terminal part of Btk by X-ray crystallography at 1.6 A resolution. This part of the kinase contains a pleckstrin homology (PH) domain and a Btk motif. The structure of the PH domain is similar to those published previously: a seven-stranded bent beta-sheet with a C-terminal alpha-helix. Individual point mutations within the Btk PH domain which cause XLA can be classified as either structural or functional in the light of the three-dimensional structure and biochemical data. All functional mutations cluster into the positively charged end of the molecule around the predicted binding site for phosphatidylinositol lipids. It is likely that these mutations inactivate the Btk pathway in cell signalling by reducing its affinity for inositol phosphates, which causes a failure in translocation of the kinase to the cell membrane. A small number of signalling proteins contain a Btk motif that always follows a PH domain in the sequence. This small module has a novel fold which is held together by a zinc ion bound by three conserved cysteines and a histidine. The Btk motif packs against the second half of the beta-sheet of the PH domain, forming a close contact with it. Our structure opens up new ways to study the role of the PH domain and Btk motif in the cellular function of Btk and the molecular basis of its dysfunction in XLA patients. PMID- 9218783 TI - The crystal structure of plant acetohydroxy acid isomeroreductase complexed with NADPH, two magnesium ions and a herbicidal transition state analog determined at 1.65 A resolution. AB - Acetohydroxy acid isomeroreductase catalyzes the conversion of acetohydroxy acids into dihydroxy valerates. This reaction is the second in the synthetic pathway of the essential branched side chain amino acids valine and isoleucine. Because this pathway is absent from animals, the enzymes involved in it are good targets for a systematic search for herbicides. The crystal structure of acetohydroxy acid isomeroreductase complexed with cofactor NADPH, Mg2+ ions and a competitive inhibitor with herbicidal activity, N-hydroxy-N-isopropyloxamate, was solved to 1.65 A resolution and refined to an R factor of 18.7% and an R free of 22.9%. The asymmetric unit shows two functional dimers related by non-crystallographic symmetry. The active site, nested at the interface between the NADPH-binding domain and the all-helical C-terminus domain, shows a situation analogous to the transition state. It contains two Mg2+ ions interacting with the inhibitor molecule and bridged by the carboxylate moiety of an aspartate residue. The inhibitor-binding site is well adjusted to it, with a hydrophobic pocket and a polar region. Only 24 amino acids are conserved among known acetohydroxy acid isomeroreductase sequences and all of these are located around the active site. Finally, a 140 amino acid region, present in plants but absent from other species, was found to make up most of the dimerization domain. PMID- 9218784 TI - Crystal structure of UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase from Escherichia coli. AB - UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase (MurD) is a cytoplasmic enzyme involved in the biosynthesis of peptidoglycan which catalyzes the addition of D glutamate to the nucleotide precursor UDP-N-acetylmuramoyl-L-alanine (UMA). The crystal structure of MurD in the presence of its substrate UMA has been solved to 1.9 A resolution. Phase information was obtained from multiple anomalous dispersion using the K-shell edge of selenium in combination with multiple isomorphous replacement. The structure comprises three domains of topology each reminiscent of nucleotide-binding folds: the N- and C-terminal domains are consistent with the dinucleotide-binding fold called the Rossmann fold, and the central domain with the mononucleotide-binding fold also observed in the GTPase family. The structure reveals the binding site of the substrate UMA, and comparison with known NTP complexes allows the identification of residues interacting with ATP. The study describes the first structure of the UDP-N acetylmuramoyl-peptide ligase family. PMID- 9218785 TI - Aerolysin and pertussis toxin share a common receptor-binding domain. AB - We have discovered that the bacterial toxins aerolysin and pertussis toxin share a common domain. This is surprising because the two toxins affect cells in very different ways. The common domain, which we call the APT domain, consists of two three-stranded antiparallel beta-sheets that come together and wrap around a central pair of helices. The APT domain shares a common fold with the C-type lectins and Link modules, and there appears to be a divergent relationship among the three families. One surface region of the APT domain is highly conserved, raising the possibility that the domains have a common function in both proteins. Mutation of one of the conserved surface residues in aerolysin, Tyr61, results in reduced receptor binding and activity, thus providing evidence that the APT domain may be involved in interaction with the toxin's receptor. Structural and biochemical evidence suggests that the APT domain contains a carbohydrate-binding site that can direct the toxins to their target cells. PMID- 9218786 TI - CD66 carcinoembryonic antigens mediate interactions between Opa-expressing Neisseria gonorrhoeae and human polymorphonuclear phagocytes. AB - Colonization of urogenital tissues by the human pathogen Neisseria gonorrhoeae is characteristically associated with purulent exudates of polymorphonuclear phagocytes (PMNs) containing apparently viable bacteria. Distinct variant forms of the phase-variable opacity-associated (Opa) outer membrane proteins mediate the non-opsonized binding and internalization of N. gonorrhoeae by human PMNs. Using overlay assays and an affinity isolation technique, we demonstrate the direct interaction between Opa52-expressing gonococci and members of the human carcinoembryonic antigen (CEA) family which express the CD66 epitope. Gonococci and recombinant Escherichia coli strains synthesizing Opa52 showed specific binding and internalization by transfected HeLa cell lines expressing the CD66 family members BGP (CD66a), NCA (CD66c), CGM1 (CD66d) and CEA (CD66e), but not that expressing CGM6 (CD66b). Bacterial strains expressing either no opacity protein or the epithelial cell invasion-associated Opa50 do not bind these CEA family members. Consistent with their different receptor specificities, Opa52 mediated interactions could be inhibited by polyclonal anti-CEA sera, while Opa50 binding was instead inhibited by heparin. Using confocal laser scanning microscopy, we observed a marked recruitment of CD66 antigen by Opa52-expressing gonococci on both the transfected cell lines and infected PMNs. These data indicate that members of the CEA family constitute the cellular receptors for the interaction with, and internalization of, N. gonorrhoeae. PMID- 9218787 TI - Identification of amino acid residues contributing to the pore of a P2X receptor. AB - P2X receptors are ion channels opened by extracellular ATP. The seven subunits currently known are encoded by different genes. It is thought that each subunit has two transmembrane domains, a large extracellular loop, and intracellular N- and C-termini, a topology which is fundamentally different from that of other ligand-gated channels such as nicotinic acetylcholine or glutamate receptors. We used the substituted cysteine accessibility method to identify parts of the molecule that form the ionic pore of the P2X2 receptor. Amino acids preceding and throughout the second hydrophobic domain (316-354) were mutated individually to cysteine, and the DNAs were expressed in HEK293 cells. For three of the 38 residues (I328C, N333C, T336C), currents evoked by ATP were inhibited by extracellular application of methanethiosulfonates of either charge (ethyltrimethylammonium, ethylsulfonate) suggesting that they lie in the outer vestibule of the pore. For two further substitutions (L338C, D349C) only the smaller ethylamine derivative inhibited the current. L338C was accessible to cysteine modification whether or not the channel was opened by ATP, but D349C was inhibited only when ATP was concurrently applied. The results indicate that part of the pore of the P2X receptor is formed by the second hydrophobic domain, and that L338 and D349 are on either side of the channel 'gate'. PMID- 9218788 TI - Tetramerization of the AKT1 plant potassium channel involves its C-terminal cytoplasmic domain. AB - All plant channels identified so far show high conservation throughout the polypeptide sequence except in the ankyrin domain which is present only in those closely related to AKT1. In this study, the architecture of the AKT1 protein has been investigated. AKT1 polypeptides expressed in the baculovirus/Sf9 cells system were found to assemble into tetramers as observed with animal Shaker-like potassium channel subunits. The AKT1 C-terminal intracytoplasmic region (downstream from the transmembrane domain) alone formed tetrameric structures when expressed in Sf9 cells, revealing a tetramerization process different from that of Shaker channels. Tests of subfragments from this sequence in the two hybrid system detected two kinds of interaction. The first, involving two identical segments (amino acids 371-516), would form a contact between subunits, probably via their putative cyclic nucleotide-binding domains. The second interaction was found between the last 81 amino acids of the protein and a region lying between the channel hydrophobic core and the putative cyclic nucleotide binding domain. As the interacting regions are highly conserved in all known plant potassium channels, the structural organization of AKT1 is likely to extend to these channels. The significance of this model with respect to animal cyclic nucleotide-gated channels is also discussed. PMID- 9218789 TI - Identification of a motor protein required for filamentous growth in Ustilago maydis. AB - The phytopathogenic fungus Ustilago maydis exists in two stages, the yeast-like haploid form and the filamentous dikaryon. Both pathogenicity and dimorphism are genetically controlled by two mating-type loci, with only the filamentous stage being pathogenic on corn. We have identified two genes (kin1 and kin2) encoding motor proteins of the kinesin family. Kin1 is most similar to the human CENP-E gene product, while Kin2 is most closely related to the conventional kinesin Nkin of Neurospora crassa. Deletion mutants of kin1 had no discernible phenotype; delta kin2 mutants, however, were severely affected in hyphal extension and pathogenicity. The wild-type dikaryon showed rapid tip growth, with all the cytoplasm being moved to the tip compartment. Left behind are septate cell wall tubes devoid of cytoplasm. In delta kin2 mutants, dikaryotic cells were formed after cell fusion, but these hyphal structures remained short and filled with cytoplasm. A functional green fluorescent protein (GFP)-Kin2 fusion was generated and used to determine the localization of the motor protein by fluorescence microscopy. Inspection of the hyphal tips by electron microscopy revealed a characteristic accumulation of darkly stained vesicles which was absent in mutant cells. We suggest that the motor protein Kin2 is involved in organizing this specialized growth zone at the hyphal tip, probably by affecting the vectorial transport of vesicles. PMID- 9218790 TI - The Aspergillus nidulans sepA gene encodes an FH1/2 protein involved in cytokinesis and the maintenance of cellular polarity. AB - Cytokinesis (septation) in the fungus Aspergillus nidulans occurs through the formation of a transient actin ring at the incipient division site. Temperature sensitive mutations in the sepA gene prevent septation and cause defects in the maintenance of cellular polarity, without affecting growth and nuclear division. The sepA gene encodes a member of the growing family of FH1/2 proteins, which appear to have roles in morphogenesis and cytokinesis in organisms such as yeast and Drosophila. Results from temperature shift and immunofluorescence microscopy experiments strongly suggest that sepA function requires a preceding mitosis and that sepA acts prior to actin ring formation. Deletion mutants of sepA exhibit temperature-sensitive growth and severe delays in septation at the permissive temperature, indicating that expression of another gene may compensate for the loss of sepA. Conidiophores formed by sepA mutants exhibit abnormal branching of the stalk and vesicle. These results suggest that sepA interacts with the actin cytoskeleton to promote formation of the actin ring during cytokinesis and that sepA is also required for maintenance of cellular polarity during hyphal growth and asexual morphogenesis. PMID- 9218791 TI - Targeting and translocation of proteins into the hydrogenosome of the protist Trichomonas: similarities with mitochondrial protein import. AB - Trichomonads are early-diverging eukaryotes that lack both mitochondria and peroxisomes. They do contain a double membrane-bound organelle, called the hydrogenosome, that metabolizes pyruvate and produces ATP. To address the origin and biological nature of hydrogenosomes, we have established an in vitro protein import assay. Using purified hydrogenosomes and radiolabeled hydrogenosomal precursor ferredoxin (pFd), we demonstrate that protein import requires intact organelles, ATP and N-ethylmaleimide-sensitive cytosolic factors. Protein import is also affected by high concentrations of the protonophore, m chlorophenylhydrazone (CCCP). Binding and translocation of pFd into hydrogenosomes requires the presence of an eight amino acid N-terminal presequence that is similar to presequences found on all examined hydrogenosomal proteins. Upon import, pFd is processed to a size consistent with cleavage of the presequence. Mutation of a conserved leucine at position 2 in the presequence to a glycine disrupts import of pFd into the organelle. Interestingly, a comparison of hydrogenosomal and mitochondrial protein presequences reveals striking similarities. These data indicate that mechanisms underlying protein targeting and biogenesis of hydrogenosomes and mitochondria are similar, consistent with the notion that these two organelles arose from a common endosymbiont. PMID- 9218792 TI - Alternative lipid remodelling pathways for glycosylphosphatidylinositol membrane anchors in Saccharomyces cerevisiae. AB - Glycosylphosphatidylinositol (GPI)-anchored membrane proteins of Saccharomyces cerevisiae exist with two types of lipid moiety--diacylglycerol or ceramide--both of which contain 26:0 fatty acids. To understand at which stage of biosynthesis these long-chain fatty acids become incorporated into diacylglycerol anchors, we compared the phosphatidylinositol moieties isolated from myo-[2-(3)H]inositol labelled protein anchors and from GPI intermediates. There is no evidence for the presence of long-chain fatty acids in any intermediate of GPI biosynthesis. However, GPI-anchored proteins contain either the phosphatidylinositol moiety characteristic of the precursor lipids or a version with a long-chain fatty acid in the sn-2 position of glycerol. The introduction of long-chain fatty acids into sn-2 occurs in the endoplasmic reticulum (ER) and is independent of the sn-2 specific acyltransferase SLC1. Analysis of ceramide anchors revealed the presence of two types of ceramide, one added in the ER and another more polar molecule which is found only on proteins which have reached the mid Golgi. In summary, the lipid of GPI-anchored proteins can be exchanged by at least three different remodelling pathways: (i) remodelling from diacylglycerol to ceramide in the ER as proposed previously; (ii) remodelling from diacylglycerol to a more hydrophobic diacylglycerol with a long-chain fatty acid in sn-2 in the ER; and (iii) remodelling to a more polar ceramide in the Golgi. PMID- 9218793 TI - Lipid remodeling leads to the introduction and exchange of defined ceramides on GPI proteins in the ER and Golgi of Saccharomyces cerevisiae. AB - Previous experiments with Saccharomyces cerevisiae had suggested that diacylglycerol-containing glycosylphosphatidylinositols (GPIs) are added to newly synthesized proteins in the endoplasmic reticulum (ER) and that ceramides subsequently are incorporated into GPI proteins by lipid remodeling. Here we prove this hypothesis by labeling yeast cells with [3H]dihydrosphingosine ([3H]DHS) and showing that this tracer is incorporated into many GPI proteins even when protein synthesis and, hence, anchor addition, is blocked by cycloheximide. [3H]DHS incorporation is greatly enhanced if endogenous synthesis of DHS is inhibited by myriocin. Labeled GPI anchors contain three types of ceramides which, based on previous and present results, are identified as DHS C26:0, phytosphingosine-C26:0 and phytosphingosine-C26:0-OH, the latter being found only on proteins which have reached the Golgi. Lipid remodeling can occur both in the ER and in a later secretory compartment. In addition, ceramide is incorporated into GPI proteins a long time after their initial synthesis by a process in which one ceramide gets replaced by another ceramide. Remodeling outside the ER requires vesicular flow from the ER to the Golgi, possibly to supply the remodeling enzymes with ceramides. PMID- 9218794 TI - Coxsackievirus protein 2B modifies endoplasmic reticulum membrane and plasma membrane permeability and facilitates virus release. AB - Digital-imaging microscopy was performed to study the effect of Coxsackie B3 virus infection on the cytosolic free Ca2+ concentration and the Ca2+ content of the endoplasmic reticulum (ER). During the course of infection a gradual increase in the cytosolic free Ca2+ concentration was observed, due to the influx of extracellular Ca2+. The Ca2+ content of the ER decreased in time with kinetics inversely proportional to those of viral protein synthesis. Individual expression of protein 2B was sufficient to induce the influx of extracellular Ca2+ and to release Ca2+ from ER stores. Analysis of mutant 2B proteins showed that both a cationic amphipathic alpha-helix and a second hydrophobic domain in 2B were required for these activities. Consistent with a presumed ability of protein 2B to increase membrane permeability, viruses carrying a mutant 2B protein exhibited a defect in virus release. We propose that 2B gradually enhances membrane permeability, thereby disrupting the intracellular Ca2+ homeostasis and ultimately causing the membrane lesions that allow release of virus progeny. PMID- 9218795 TI - Minimal requirements for calcium oscillations driven by the IP3 receptor. AB - Hormones and neurotransmitters that act through inositol 1,4,5-trisphosphate (IP3) can induce oscillations of cytosolic Ca2+ ([Ca2+]c), which render dynamic regulation of intracellular targets. Imaging of fluorescent Ca2+ indicators located within intracellular Ca2+ stores was used to monitor IP3 receptor channel (IP3R) function and to demonstrate that IP3-dependent oscillations of Ca2+ release and re-uptake can be reproduced in single permeabilized hepatocytes. This system was used to define the minimum essential components of the oscillation mechanism. With IP3 clamped at a submaximal concentration, coordinated cycles of IP3R activation and subsequent inactivation were observed in each cell. Cycling between these states was dependent on feedback effects of released Ca2+ and the ensuing [Ca2+]c increase, but did not require Ca2+ re-accumulation. [Ca2+]c can act at distinct stimulatory and inhibitory sites on the IP3R, but whereas the Ca2+ release phase was driven by a Ca2+-induced increase in IP3 sensitivity, Ca2+ release could be terminated by intrinsic inactivation after IP3 bound to the Ca2+ sensitized IP3R without occupation of the inhibitory Ca2+-binding site. These findings were confirmed using Sr2+, which only interacts with the stimulatory site. Moreover, vasopressin induced Sr2+ oscillations in intact cells in which intracellular Ca2+ was completely replaced with Sr2+. Thus, [Ca2+]c oscillations can be driven by a coupled process of Ca2+-induced activation and obligatory intrinsic inactivation of the Ca2+-sensitized state of the IP3R, without a requirement for occupation of the inhibitory Ca2+-binding site. PMID- 9218797 TI - Acceleration of intracellular targeting of antigen by the B-cell antigen receptor: importance depends on the nature of the antigen-antibody interaction. AB - The B-cell antigen receptor (BCR) internalizes bound antigen such that antigen derived peptides become associated with emigrating major histocompatibility complex (MHC) class II molecules for presentation to T cells. Experiments with B cell transfectants reveal that BCR confers a specificity of intracellular targeting since chimeric antigen receptors which internalize antigen by virtue of a heterologous cytoplasmic domain do not necessarily give rise to presentation. In contrast, however, previous studies have shown that antigen binding to irrelevant cell surface molecules (e.g. transferrin receptor, MHC class I) can ultimately lead to presentation. The solution to this paradox appears to be that the intracellular targeting by BCR actually reflects an acceleration of antigen delivery. Depending on the nature of the BCR-antigen interaction, this accelerated targeting can be essential in determining whether or not internalization leads to significant presentation. Physiologically, the accelerated delivery of antigen by BCR could prove of particular importance early in the immune response when antigen-BCR interaction is likely to be poor. PMID- 9218796 TI - Modulation of murine melanocyte function in vitro by agouti signal protein. AB - Molecular and biochemical mechanisms that switch melanocytes between the production of eumelanin or pheomelanin involve the opposing action of two intercellular signaling molecules, alpha-melanocyte-stimulating hormone (MSH) and agouti signal protein (ASP). In this study, we have characterized the physiological effects of ASP on eumelanogenic melanocytes in culture. Following exposure of black melan-a murine melanocytes to purified recombinant ASP in vitro, pigmentation was markedly inhibited and the production of eumelanosomes was decreased significantly. Melanosomes that were produced became pheomelanosome like in structure, and chemical analysis showed that eumelanin production was significantly decreased. Melanocytes treated with ASP also exhibited time- and dose-dependent decreases in melanogenic gene expression, including those encoding tyrosinase and tyrosinase-related proteins 1 and 2. Conversely, melanocytes exposed to MSH exhibited an increase in tyrosinase gene expression and function. Simultaneous addition of ASP and MSH at approximately equimolar concentrations produced responses similar to those elicited by the hormone alone. These results demonstrate that eumelanogenic melanocytes can be induced in culture by ASP to exhibit features characteristic of pheomelanogenesis in vivo. Our data are consistent with the hypothesis that the effects of ASP on melanocytes are not mediated solely by inhibition of MSH binding to its receptor, and provide a cell culture model to identify novel factors whose presence is required for pheomelanogenesis. PMID- 9218798 TI - Activation of the novel stress-activated protein kinase SAPK4 by cytokines and cellular stresses is mediated by SKK3 (MKK6); comparison of its substrate specificity with that of other SAP kinases. AB - A cDNA was cloned that encodes human stress-activated protein kinase-4 (SAPK4), a novel MAP kinase family member whose amino acid sequence is approximately 60% identical to that of the other three SAP kinases which contain a TGY motif in their activation domain. The mRNA encoding SAPK4 was found to be widely distributed in human tissues. When expressed in KB cells, SAPK4 was activated in response to cellular stresses and pro-inflammatory cytokines, in a manner similar to other SAPKs. SAPK4 was activated in vitro by SKK3 (also called MKK6) or when co-transfected with SKK3 into COS cells. SKK3 was the only activator of SAPK4 that was induced when KB cells were exposed to a cellular stress or stimulated with interleukin-1. These findings indicate that SKK3 mediates the activation of SAPK4. The substrate specificity of SAPK4 in vitro was similar to that of SAPK3. Both enzymes phosphorylated the transcription factors ATF2, Elk-1 and SAP-1 at similar rates, but were far less effective than SAPK2a (also called RK/p38) or SAPK2b (also called p38beta) in activating MAPKAP kinase-2 and MAPKAP kinase-3. Unlike SAPK1 (also called JNK), SAPK3 and SAPK4 did not phosphorylate the activation domain of c-Jun. Unlike SAPK2a and SAPK2b, SAPK4 and SAPK3 were not inhibited by the drugs SB 203580 and SB 202190. Our results suggest that cellular functions previously attributed to SAPK1 and/or SAPK2 may be mediated by SAPK3 or SAPK4. PMID- 9218799 TI - A role for the divergent actin gene, ACT2, in nuclear pore structure and function. AB - We have identified a temperature-sensitive allele of the yeast divergent actin gene ACT2, act2-1, which displays defects in nuclear pore complex (NPC) structure and nuclear import at the restrictive temperature. Although defective in nuclear import, act2-1 cells still selectively retain reporter proteins in the nucleus, and by indirect immunofluorescence the actin cytoskeleton appears normal. Previous studies in Acanthamoeba and Saccharomyces cerevisiae reported that the cellular location of Act2p partially overlaps that of conventional actin, indicating that it has a cytoskeletal function. In this study, both immunofluorescence localization and cellular fractionation of different epitope tagged versions of Act2p also reveal an association with the nucleus, suggesting an independent nuclear function for Act2p. Analysis of act2-1 by electron microscopy, 30 min after a shift to the restrictive temperature (37 degrees C), reveals a striking aberration in NPC morphology; NPCs appear as abnormal densities on either side of, rather than spanning, the nuclear envelope. Immunoelectron microscopy confirms that these densities contain XFXFG nucleoporins. act2-1 is synthetically lethal in combination with a deletion in the XFXFG nucleoporin gene, NUP1, or a mutation in the nuclear localization sequence receptor gene, SRP1. Act2p and Srp1p co-immunoprecipitate, suggesting that the proteins exist in a complex. Together our data argue that Act2p plays an important role in NPC structure and function. PMID- 9218801 TI - High level transcription of a member of a repeated gene family confers dehydration tolerance to callus tissue of Craterostigma plantagineum. AB - An experimental system has been developed which allows the identification of intermediates in the abscisic acid (ABA) signal transduction pathway leading to desiccation tolerance in plants. Desiccation tolerance in callus of the resurrection plant Craterostigma plantagineum is mediated via the plant hormone ABA, which induces the expression of gene products related to desiccation tolerance. Based on T-DNA activation tagging, a gene (CDT-1) was isolated which encodes a signalling molecule in the ABA transduction pathway. Constitutive overexpression of CDT-1 leads to desiccation tolerance in the absence of ABA and to the constitutive expression of characteristic transcripts. CDT-1 represents a novel gene with unusual features in its primary sequence. The CDT-1 gene resembles in several features SINE retrotransposons. Mechanisms by which CDT-1 activates the pathway could be via a regulatory RNA or via a short polypeptide. PMID- 9218800 TI - The K nuclear shuttling domain: a novel signal for nuclear import and nuclear export in the hnRNP K protein. AB - Protein import into the nucleus and export from the nucleus are signal-mediated processes that require energy. The nuclear transport process about which the most information is currently available is classical nuclear localization signal (NLS) mediated nuclear import. However, details concerning the signal-mediated export of proteins and RNAs as well as alternative nuclear import pathways are beginning to emerge. An example of this is the heterogeneous nuclear ribonucleoprotein (hnRNP) A1 protein which, by virtue of its M9 domain, is actively exported from the nucleus and imported into the nucleus via a novel pathway mediated by the recently characterized transportin protein. Here we report that the shuttling hnRNP K protein contains a novel shuttling domain (termed KNS) which has many of the characteristics of M9, in that it confers bi-directional transport across the nuclear envelope. KNS-mediated nuclear import is dependent on RNA polymerase II transcription, and we show that a classical NLS can override this effect. Furthermore, KNS accesses a separate import pathway from either classical NLSs or M9. This demonstrates the existence of a third protein import pathway into the nucleus and thereby defines a new type of nuclear import/export signal. PMID- 9218802 TI - Regulation of NFKB1 proteins by the candidate oncoprotein BCL-3: generation of NF kappaB homodimers from the cytoplasmic pool of p50-p105 and nuclear translocation. AB - The candidate oncoprotein BCL-3 has been shown to function as a transcriptional co-activator for homodimers of NF-kappaB p50 and p50B. We expressed BCL-3 ectopically in pro-B cell lines and found that these cells exhibited a dramatic increase in nuclear kappaB motif binding activity of p50 homodimers containing BCL-3 in the complex. Co-transfection and in vitro reconstitution experiments revealed that the complex of p50 with its precursor p105 (p50-p105), which was shown to accumulate in the cytoplasm of the pro-B cell lines, is required for induction of DNA binding of p50 homodimers by BCL-3. However, we could see no in vivo or in vitro evidence of a BCL-3-induced increase in proteolytic processing. Instead, BCL-3-mediated reorganization of NFKB1 subunits was demonstrated in vitro. Immunofluorescence staining clearly demonstrated that the transition from cytoplasmic p50-p105 to nuclear p50 homodimers was induced by BCL-3 expression. Thus BCL-3 has versatile functions: cytoplasmic activation of p50 homodimers, their nuclear translocation and, as previously shown, modulation of the transcriptional machinery in the nucleus. PMID- 9218803 TI - Co-localization of Polycomb protein and GAGA factor on regulatory elements responsible for the maintenance of homeotic gene expression. AB - The Polycomb group and trithorax group genes of Drosophila are required for maintaining the differential expression state of developmental regulators, such as the homeotic genes, in a stable and heritable manner throughout development. The Polycomb group genes have been suggested to act by regulating higher order chromatin and packaging repressed chromosomal domains in a heterochromatin-like structure. We have mapped, at high resolution, the distribution of Polycomb protein on the bithorax complex of Drosophila tissue culture cells, using an improved formaldehyde cross-linking and immunoprecipitation technique. Polycomb protein is not distributed homogeneously on the regulatory regions of the repressed Ultrabithorax and abdominal-A genes, but is highly enriched at discrete sequence elements, many of which coincide with previously mapped Polycomb group response elements (PREs). Our results further suggest that Polycomb protein spreads locally over a few kilobases of DNA surrounding PREs, perhaps to stabilize silencing complexes. GAGA factor/Trithorax-like, a member of the trithorax group, is also bound at those PREs which contain GAGA consensus-binding sites. Two modes of binding can be distinguished: a high level binding to elements in the regulatory domain of the expressed Abdominal-B gene, and a low level of binding to Polycomb-bound PREs in the inactive domains of the bithorax complex. We propose that GAGA factor binds constitutively to regulatory elements in the bithorax complex, which function both as PREs and as trithorax group response elements. PMID- 9218804 TI - Transcriptional termination signals for RNA polymerase II in fission yeast. AB - Transcription 'run-on' (TRO) analysis using permeabilized yeast cells indicates that transcription terminates between 180 and 380 bp downstream of the poly(A) site of the Schizosaccharomyces pombe ura4 gene. Two signals direct RNA polymerase II (pol II) to stop transcription: the previously identified 3' end formation signals located close to the poly(A) site and an additional downstream element (DSE) located at the region of termination. The downstream signal (135 bp) appears to act by pausing the elongating polymerase. TRO analysis indicates that elevated levels of transcribing polymerases accumulate over the DSE and that removal of this signal leads to transcription proceeding beyond the normal termination region. Furthermore, when inserted between two competing polyadenylation signals, this DSE increases the utilization of upstream poly(A) sites in vivo. We show that polymerase pausing over an extended region of template ensures termination of pol II transcription close to the poly(A) site. PMID- 9218805 TI - Functional dissection of a transcriptionally active, target-specific Hox-Pbx complex. AB - Hox genes control cell fates and specify regional identities in vertebrate development. Hox proteins show a relaxed DNA-binding selectivity in vitro, suggesting that functional specificity is achieved in vivo through the action of transcriptional co-factors. Pbx proteins are good candidates for such a role, on the basis of both genetic and biochemical evidence. We report that the human Pbx1 and HOXB1 proteins can cooperatively activate transcription through a genetically characterized Hox target, i.e. an autoregulatory element directing spatially restricted expression of the murine Hoxb-1 gene (b1-ARE) in the developing hindbrain. On the b1-ARE, only a restricted subset of HOX proteins (HOXA1, HOXB1, HOXA2) are able to bind cooperatively with Pbx1 and activate transcription. Selective recognition of the b1-ARE is mediated by the N-terminal region of the HOX homeodomain. The DNA-binding and protein-protein interaction functions of HOXB1 and Pbx1 are all necessary for the assembly of a transcriptionally active complex on the b1-ARE. Functional dissection of the complex allowed the localization of the main activation domain in the HOXB1 N-terminal region, and of an additional one in the C-terminal region of Pbx1 contained in the Pbx1a but not in the alternatively spliced Pbx1b isoform. Our results indicate that Pbx1 acts as a transcriptional co-factor of Hox proteins, allowing selective recognition and cooperative activation of regulatory target sequences. PMID- 9218806 TI - FIS activates sequential steps during transcription initiation at a stable RNA promoter. AB - FIS (factor for inversion stimulation) is a small dimeric DNA-bending protein which both stimulates DNA inversion and activates transcription at stable RNA promoters in Escherichia coli. Both these processes involve the initial formation of a complex nucleoprotein assembly followed by local DNA untwisting at a specific site. We have demonstrated previously that at the tyrT promoter three FIS dimers are required to form a nucleoprotein complex with RNA polymerase. We now show that this complex is structurally dynamic and that FIS, uniquely for a prokaryotic transcriptional activator, facilitates sequential steps in the initiation process, enabling efficient polymerase recruitment, untwisting of DNA at the transcription startpoint and finally the escape of polymerase from the promoter. Activation of all these steps requires that the three FIS dimers bind in helical register. We suggest that FIS acts by stabilizing a DNA microloop whose topology is coupled to the local topological transitions generated during the initiation of transcription. PMID- 9218807 TI - Repressor induced site-specific binding of HU for transcriptional regulation. AB - Transcription from two overlapping gal promoters is repressed by Gal repressor binding to bipartite gal operators, O(E) and O(I), which flank the promoters. Concurrent repression of the gal promoters also requires the bacterial histone like protein HU which acts as a co-factor. Footprinting experiments using iron EDTA-coupled HU show that HU binding to gal DNA is orientation specific and is specifically dependent upon binding of GalR to both O(E) and O(I). We propose that HU, in concert with GalR, forms a specific nucleoprotein higher order complex containing a DNA loop. This way, HU deforms the promoter to make the latter inactive for transcription initiation while remaining sensitive to inducer. The example of gal repression provides a model for studying how a 'condensed' DNA becomes available for transcription. PMID- 9218808 TI - Consistent gene silencing in transgenic plants expressing a replicating potato virus X RNA. AB - Tobacco plants were transformed with constructs in which the transgene was a cDNA of replicating potato virus X (PVX) RNA. The constructs, referred to here as amplicons, were the intact genome of PVX and PVX constructs modified to carry the beta-glucuronidase (GUS) reporter gene either as an additional gene or as a replacement for the coat protein gene (PVX/GUS/CP and PVX/GUS respectively). Transformed plants carrying these constructs displayed several phenotypes that we attribute to post-transcriptional gene silencing. These phenotypes include the absence of viral symptoms, low accumulation of transgene-derived RNA, extreme strain-specific resistance against PVX, low and non-uniform GUS expression (in the PVX/GUS and PVX/GUS/CP plants) and suppression of transiently expressed RNA sharing homology with the transgene. Importantly, the amplicon-mediated gene silencing was exhibited in all lines tested. There was no evidence of gene silencing in seven lines expressing a PVX RNA that was unable to replicate. From these data we conclude that the replicating viral RNA is a potent trigger of gene silencing. Moreover, amplicon-mediated gene silencing provides an important new strategy for the consistent activation of gene silencing in transgenic plants. PMID- 9218809 TI - The structure of the isolated, central hairpin of the HDV antigenomic ribozyme: novel structural features and similarity of the loop in the ribozyme and free in solution. AB - The structure of an RNA hairpin containing a seven-nucleotide loop that is present in the self-cleaving sequence of hepatitis delta virus antigenomic RNA was determined by high resolution NMR spectroscopy. The loop, which is composed of only one purine and six pyrimidines, has a suprisingly stable structure, mainly supported by sugar hydroxyl hydrogen bonds and base-base and base phosphate stacking interactions. Compared with the structurally well-determined, seven-membered anticodon loop in tRNA, the sharp turn which affects the required 180 degrees change in direction of the sugar-phosphate backbone in the loop is shifted one nucleotide in the 3' direction. This change in direction can be characterized as a reversed U-turn. It is expected that the reversed U-turn may be found frequently in other molecules as well. There is evidence for a new non Watson-Crick UC base pair formed between the first and the last residue in the loop, while most of the other bases in the loop are pointing outwards making them accessible to solvent. From chemical modification, mutational and photocrosslinking studies, a similar picture develops for the structure of the hairpin in the active ribozyme indicating that the loop structure in the isolated hairpin and in the ribozyme is very similar. PMID- 9218810 TI - Rapamycin suppresses 5'TOP mRNA translation through inhibition of p70s6k. AB - Treatment of mammalian cells with the immunosuppressant rapamycin, a bacterial macrolide, selectively suppresses mitogen-induced translation of an essential class of mRNAs which contain an oligopyrimidine tract at their transcriptional start (5'TOP), most notably mRNAs encoding ribosomal proteins and elongation factors. In parallel, rapamycin blocks mitogen-induced p70 ribosomal protein S6 kinase (p70s6k) phosphorylation and activation. Utilizing chimeric mRNA constructs containing either a wild-type or disrupted 5'TOP, we demonstrate that an intact polypyrimidine tract is required for rapamycin to elicit an inhibitory effect on the translation of these transcripts. In turn, a dominant-interfering p70s6k, which selectively prevents p70s6k activation by blocking phosphorylation of the rapamycin-sensitive sites, suppresses the translation of the chimeric mRNA containing the wild-type but not the disrupted 5'TOP. Conversion of the principal rapamycin-sensitive p70s6k phosphorylation site, T389, to an acidic residue confers rapamycin resistance on the kinase and negates the inhibitory effects of the macrolide on 5'TOP mRNA translation in cells expressing this mutant. The results demonstrate that the rapamycin block of mitogen-induced 5'TOP mRNA translation is mediated through inhibition of p70s6k activation. PMID- 9218812 TI - Zepp, a LINE-like retrotransposon accumulated in the Chlorella telomeric region. AB - Six copies of insertion elements accumulate in the subtelomeric region immediately proximal to the telomeric repeats on Chlorella chromosome I. The elements, designated Zepps, bear the characteristic features of non-viral (LINE like) retrotransposons, including a poly(A) tail, 5'-truncations, a retroviral reverse transcriptase-like ORF and flanking target duplications. Detailed sequence analysis of the Chlorella subtelomeric region revealed a novel mechanism of Zepp transposition; successive insertions of each Zepp element into another Zepp as a target, leaving a tandem array of their 3'-regions with poly(A) tracts facing toward the centromere. Only the most distal Zepp copy was inverted to connect its poly(A) tail with the telomeric repeats. A similar Zepp cluster but without the telomeric repeats was also found at the terminus of another Chlorella chromosome. These structures contrast with that proposed for the addition of HeT A and TART elements to Drosophila telomeres. Expression of Zepp elements is induced by heat shock treatment. Possible roles of the subtelomeric retrotransposons in formation and maintenance of telomeres are discussed. PMID- 9218811 TI - The terminal DNA structure of mammalian chromosomes. AB - In virtually all eukaryotic organisms, telomeric DNA is composed of a variable number of short direct repeats. While the primary sequence of telomeric repeats has been determined for a great variety of species, the actual physical DNA structure at the ends of a bona fide metazoan chromosome with a centromere is unknown. It is shown here that an overhang of the strand forming the 3' ends of the chromosomes, the G-rich strand, is found at mammalian chromosome ends. Moreover, on at least some telomeres, the overhangs are > or = 45 bases long. Such surprisingly long overhangs were present on chromosomes derived from fully transformed tissue culture cells and normal G0-arrested peripheral leukocytes. Thus, irrespective of whether the cells were actively dividing or arrested, a very similar terminal DNA arrangement was found. These data suggest that the ends of mammalian and possibly all vertebrate chromosomes consist of an overhang of the G-rich strand and that these overhangs may be considerably larger than previously anticipated. PMID- 9218813 TI - Negative control of DNA replication by hydrolysis of ATP bound to DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli. AB - DnaA protein, the initiation factor for chromosomal DNA replication in Escherichia coli, is activated by ATP. ATP bound to DnaA protein is slowly hydrolyzed to ADP, but the physiological role of ATP hydrolysis is unclear. We constructed, by site-directed mutagenesis, mutated DnaA protein with lower ATPase activity, and we examined its function in vitro and in vivo. The ATPase activity of purified mutated DnaA protein (Glu204-->Gln) decreased to one-third that of the wild-type DnaA protein. The mutation did not significantly affect the affinity of DnaA protein for ATP or ADP. The mutant dnaA gene showed lethality in wild-type cells but not in cells growing independently of the function of oriC. Induction of the mutated DnaA protein in wild-type cells caused an overinitiation of DNA replication. Our results lead to the thesis that the intrinsic ATPase activity of DnaA protein negatively regulates chromosomal DNA replication in E. coli cells. PMID- 9218814 TI - Action of site-specific recombinases XerC and XerD on tethered Holliday junctions. AB - In Xer site-specific recombination, two related recombinases, XerC and XerD, mediate the formation of recombinant products using Holliday junction-containing DNA molecules as reaction intermediates. Each recombinase catalyses the exchange of one pair of specific strands. By using synthetic Holliday junction-containing recombination substrates in which two of the four arms are tethered in an antiparallel configuration by a nine thymine oligonucleotide, we show that XerD catalyses efficient strand exchange only when its substrate strands are 'crossed'. XerC also catalyses very efficient strand exchange when its substrate strands are 'crossed', though it also appears to be able to mediate strand exchange when its substrate strands are 'continuous'. By using chemical probes of Holliday junction structure in the presence and absence of bound recombinases, we show that recombinase binding induces unstacking of the bases in the centre of the recombination site, indicating that the junction branch point is positioned there and is distorted as a consequence of recombinase binding. PMID- 9218815 TI - The isomeric preference of Holliday junctions influences resolution bias by lambda integrase. AB - Lambda site-specific recombination proceeds by a pair of sequential strand exchanges that first generate and then resolve a Holliday junction intermediate. A family of synthetic Holliday junctions with the branch point constrained to the center of the 7 bp overlap region was used to show that resolution of the top strands and resolution of the bottom strands are symmetrical but stereochemically distinct processes. Lambda integrase is sensitive to isomeric structure, preferentially resolving the pair of strands that are crossed in the protein-free Holliday junction. At the branch point of stacked immobile Holliday junctions, the number of purines is preferentially maximized in the crossed (versus continuous) strands if there is an inequality of purines between strands of opposite polarity. This stacking preference was used to anticipate the resolution bias of freely mobile junctions and thereby to reinforce the conclusions with monomobile junctions. The results provide a strong indication that in the complete recombination reaction a restacking of helices occurs between the top and bottom strand exchanges. PMID- 9218816 TI - Are anti-ribosomal P protein antibodies a type of anti-lymphocyte antibody? PMID- 9218817 TI - Synovial fluid T cell clones from oligoarticular juvenile arthritis patients display a prevalent Th1/Th0-type pattern of cytokine secretion irrespective of immunophenotype. AB - The aim of the present study was to investigate the patterns of cytokine production by T cell clones raised from in vivo activated synovial fluid (SF) mononuclear cells (MNC) of five patients with oligoarticular juvenile arthritis (JA). Freshly isolated SF T cells were cultured in vitro with low dose recombinant IL-2 and subsequently cloned by limiting dilution. Sixty-four clones were obtained from the five patients studied. Fifty-nine clones were TCR alpha/beta+, either CD4+ (n = 43) or CD8+ (n = 15). The remaining five clones were TCR gamma/delta+, CD4-, CD8-. Clone immunophenotypes differed in the individual patients. Forty-four T cell clones were stimulated with phytohaemagglutinin (PHA) and phorbol myristate acetate (PMA) and supernatants tested for the presence of IL-2, IL-4, IL-5 and interferon-gamma (IFN-gamma) by ELISA or bioassays. Cytokine mRNA accumulation was tested by reverse transcriptase-polymerase chain reaction (RT-PCR). Most of 44 clones tested released large amounts of IFN-gamma irrespective of the immunophenotype. Of these, 27 were classified as Th1-type and 17 as Th0-type based upon the IFN gamma/IL-4 ratio in culture supernatants. Finally, when 10 representative T cell clones were tested for pro- and anti-inflammatory cytokines, gene expression by RT-PCR, all of them were found to express the granulocyte-macrophage colony stimulating factor (GM-CSF), tumour necrosis factor-alpha (TNF-alpha), IL-10 and transforming growth factor-beta 1 (TGF-beta1) genes, and half of them IL-6 and IL 8 mRNA. In conclusion, T cell clones, that represent the progeny of in vivo activated SF T cells from oligoarticular JA patients, display heterogeneous immunophenotypes, but all share the ability to produce large amounts of IFN gamma, with a predominant Th1/Th0 pattern. The expression of pro- and anti inflammatory cytokine genes in these clones suggests that in vivo activated SF T cells modulate joint inflammation in a complex fashion. PMID- 9218818 TI - Anti-ribosomal and 'P-peptide'-specific autoantibodies bind to T lymphocytes. AB - Patients with systemic lupus erythematosus (SLE) frequently have anti-lymphocyte autoantibodies, some of which also bind to surfaces of neurons. Since anti ribosomal P protein autoantibodies (anti-P) from SLE patients also bind to surfaces of neurons, we hypothesized that anti-P are anti-lymphocyte antibodies. A panel of human T lymphocytes was evaluated for anti-P binding by indirect immunofluorescence. Affinity-purified anti-ribosomal antibodies were used as a source of anti-P. These autoantibodies bound to the surfaces of all transformed T cell lines tested. This binding was not mediated by Fc receptors. It was inhibitable by ribosomes. Anti-P bound to circulating T lymphocytes from healthy adults and children. They also bound to thymocytes and cord blood T cells from normal neonates. Circulating T cells from SLE patients with anti-P bound less anti-P than cells from healthy controls. Two patients were studied on multiple occasions. The capacity of their T cells to bind anti-P correlated inversely with titres of anti-ribosomal antibodies. Anti-ribosomal antibodies, other than anti P, also appear to bind to T cells. The surface of T cells contains a protein with the size and antigenicity of the ribosomal P protein, P0. We conclude that anti ribosomal antibodies are a subset of anti-lymphocyte autoantibodies. Their possible role in the pathogenesis of lymphopenia or lymphocyte dysfunction in SLE has to be defined in further studies. PMID- 9218819 TI - Reduced protein tyrosine phosphatase (PTPase) activity of CD45 on peripheral blood lymphocytes in patients with systemic lupus erythematosus (SLE). AB - To disclose the mechanism of aberrant function of peripheral blood lymphocytes (PBL) in SLE, we focused on the catalytic function of CD45, and determined the CD45 PTPase activity in SLE patients. The sample population consisted of 32 SLE patients with different disease activity. PTPase activity of cell lysates immunoprecipitated by anti-CD45 MoAb was assayed against phosphotyrosine analogue PNPP, followed by measuring the release of para-nitro phenol at 410 nm. CD45 PTPase activity of PBL was significantly decreased in SLE patients, compared with that of normal controls and patients with systemic sclerosis (964 +/- 265, 1202 +/- 172, 1210 +/- 125, respectively; SLE versus normal, P<0.05). It was correlated with SLE Disease Activity Index (SLEDAI) score (r = 0.597, P = 0.0006), but not with the dose of prednisolone (r = 0.214, P = 0.2657), indicating that CD45 PTPase activity became reduced when the disease was active, but it was not affected by prednisolone. Moreover, it was not corrected by in vitro culture with or without stimulation. The expression of CD45 on PBL was comparable between normal and SLE, raising a possibility that it may be due to aberrant regulation of catalytic function of CD45 in SLE. Given the evidence that tyrosine phosphorylation of cellular proteins by tyrosine kinases and phosphatases is one of the key biochemical events in the signal transduction pathway, the decreased CD45 PTPase activity in SLE may account for the defective signal transduction via TCR/CD3, leading to dysregulated effector function of the lymphocytes. PMID- 9218820 TI - Specificity and immunochemical properties of antibodies to bacterial DNA in sera of normal human subjects and patients with systemic lupus erythematosus (SLE). AB - To elucidate the mechanisms of anti-DNA production, we assessed the binding of sera of normal human subjects (NHS) and patients with SLE to a panel of bacterial and mammalian DNA. Using single-stranded DNA as antigens in an ELISA, NHS showed significant binding to some but not all bacterial DNA, while lacking reactivity to calf thymus DNA. Among bacterial DNA, the highest levels of binding were observed with DNA from Micrococcus lysodeikticus and Staphylococcus aureus. In contrast, SLE sera showed high levels of binding to all DNA tested. To evaluate further immunochemical properties of the anti-DNA antibodies, the subclass distribution of these responses was evaluated by subclass-specific reagents. While NHS showed a predominance of IgG2 antibodies to bacterial DNA, SLE sera had a predominance of IgG1 antibodies to these antigens. Together, these results provide further evidence for the antigenicity of bacterial DNA and suggest that NHS and SLE anti-DNA differ in the patterns of epitope recognition as well as mechanisms of induction. PMID- 9218821 TI - Anti-Ro52 antibodies frequently co-occur with anti-Jo-1 antibodies in sera from patients with idiopathic inflammatory myopathy. AB - We analysed 112 idiopathic inflammatory myopathy (IIM) sera for the presence of anti-Ro, anti-La and anti-histidyl-tRNA synthetase (Jo-1) autoantibodies, and subsequently mapped B cell epitopes on the Ro52 protein recognized by anti-Ro52+ IIM sera. Sera were characterized by immunoblotting, ELISA and RNA precipitation. Both anti-Ro60 and anti-La activity was found in 4% of IIM sera. Anti-Ro52 antibodies were present in 20% of IIM sera. However, in anti-Jo-1+ IIM sera (21%), the frequency of the anti-Ro52 antibodies was found to be much higher (58%). No cross-reactivity between anti-Ro52 and anti-Jo-1 antibodies could be detected in these sera. To learn more about the nature of anti-Ro52 antibodies occurring in IIM sera, we analysed the major epitopes of the Ro52 protein targeted by anti-Ro52+ IIM sera by immunoprecipitation of in vitro translated Ro52 deletion mutants. The major epitope was mapped in the region bordered by amino acids 126 and 252. This part of the protein includes a long alpha-helical region which contains two potential coiled-coil domains as well as a leucine zipper motif. Although no difference in Ro52 epitope recognition between anti-Jo 1+ and anti-Jo-1- IIM sera could be observed, our results suggest that the autoimmune response against Ro52 and Jo-1 in IIM patients is coupled. PMID- 9218822 TI - Role of tumour necrosis factor-alpha (TNF-alpha) in the induction of HIV-1 gp120 mediated CD4+ T cell anergy. AB - The HIV-1 envelope glycoprotein (gp 120) is known to induce antigen-specific and non-specific CD4+ T cell anergy. We found that early T cell activation, as indicated by HLA-DP expression in the early G1 (G1A) phase of the cell cycle, and the inhibition of mitogen-mediated IL-2 production induced by gp120, required TNF alpha produced by gp120-stimulated macrophages. In the presence of an antibody to TNF-alpha, these changes induced by gp120 were inhibited, while recombinant TNF alpha induced similar abnormalities of CD4+ T cells, even in the absence of gp120. On the other hand, inhibition of the mixed lymphocyte reaction (MLR) in CD4+ T cells by gp120, which may be related to gp120-mediated down-regulation of CD4 expression on T cells and activation of protein tyrosine kinase p56(lck) in CD4+ T cells, was observed even in the absence of macrophage-derived TNF-alpha induced by gp120. These observations indicate that both TNF-alpha-dependent and independent events contribute to gp120-mediated CD4+ T cell anergy, and TNF-alpha appears to play an important role in inducing CD4+ T cell anergy in HIV-1 infection. PMID- 9218823 TI - Quantification of IgA and IgG and specificities of antibodies to viral proteins in parotid saliva at different stages of HIV-1 infection. AB - Paired sera and parotid saliva from 75 HIV-1-infected patients, divided in three equal groups with CD4+ cell counts > 500, 200-500 and < 200/mm3, respectively, were analysed for IgG, IgA and secretory IgA (sIgA) concentrations and for IgG and IgA antibody directed to HIV-1. Twenty-nine age-matched HIV-subjects were used as controls. In serum the concentrations of immunoglobulins were significantly increased in HIV-infected subjects compared with controls, and a progressive increase of IgA and sIgA was noticed while the CD4+ cell count decreased. In contrast, concentrations of IgA and sIgA were not different in parotid saliva between the four subject groups. By an ELISA test directed towards HIV-1 proteins, 73 of the 75 serum specimens from the HIV-infected subjects (97%) and 43 of the corresponding saliva (57%) were found positive for specific IgA antibodies to HIV-1, with an even distribution among the three groups of patients. By Western blotting multiple specificities of IgA to HIV-1 proteins were not frequently found in patients. By contrast, in spite of an IgG concentration in saliva about 100 times lower than that of IgA, reactivities were significantly higher for IgG than for IgA antibodies, especially to env and to pol HIV-1 products. Altogether, these data suggest that the regulation of IgA production in HIV-infected subjects is independent in serum and in parotid saliva. This imbalance of IgA/IgG antibodies to HIV-1 at the mucosal level appears to be a specific feature of HIV-1 infection, and may raise important issues in terms of local protection after immunization. PMID- 9218824 TI - The IL-1 system in HIV infection: peripheral concentrations of IL-1beta, IL-1 receptor antagonist and soluble IL-1 receptor type II. AB - The proinflammatory cytokine IL-1beta is thought to be involved in ongoing HIV disease. Furthermore, its naturally occurring inhibitors soluble IL-1 receptor type II (sIL-1RII) and IL-1 receptor antagonist (IL-1Ra) may play a pivotal role in regulating its biological action. To investigate the involvement of the IL-1 system we determined serum levels of IL-1beta, IL-1Ra and sIL-1RII in 90 HIV- patients. The obtained values were compared with markers of disease progression such as CD4+ count, 5'-neopterin. Beta2-microglobulin and soluble tumour necrosis factor receptors (sTNF-R) p55 and p75 and then compared with C-reactive protein (CRP), granulocyte count, IL-6 and TNF-alpha. While IL-1Ra concentrations increased significantly with progressive CDC disease stages, sIL-1RII and IL 1beta were not altered in our cohort. IL-1Ra showed statistical relation to decreasing CD4+ lymphocytes and increasing 5'-neopterin, beta2-microglobulin, sTNF-R p55, sTNF-R p75. Furthermore, IL-1Ra correlated positively with serum IL 6, TNF-alpha, CRP and granulocytes. In contrast, sIL-1RII and IL-1beta tended to show an inverse correlation or showed no significant relationship to all these parameters. IL-1beta was measurable only in a limited number of samples. IL-1Ra showed a clear relationship to acute inflammatory events as well as to the different disease stages. Our data suggest a dissociation between IL-1Ra and sIL 1RII serum levels which may indicate that the two IL-1 binding proteins have different pathophysiological roles in HIV infection. PMID- 9218825 TI - Naive and memory T cell infiltrates in chronic hepatitis C: phenotypic changes with interferon treatment. AB - The phenotypes of infiltrating lymphocytes in liver with chronic hepatitis C, including changes associated with interferon (IFN) treatment, were characterized. Specimens obtained from 22 patients treated with IFN were examined using avidin biotin-peroxidase immunohistochemistry. In areas of lobular and periportal inflammation, most lymphocytes were CD8+ T cells of the CD45RO+ (memory) subset. The centres of lymphoid follicles were occupied by CD20+ B cells and a few CD4+ T cells which were CD45RA+ (naive subset). Follicular centres were surrounded mainly with CD4+ T cells. CD8+ T cells, mostly CD45RO+, were scattered through the mantle zones of follicles and extended around them. No significant changes in CD45RA+ lobular infiltrates accompanied IFN treatment. On the other hand, the number of CD45RO+ lobular infiltrates decreased after IFN treatment in complete responders (P < 0.01). Moreover, there were significant correlations between CD45RO+ cell counts and serum alanine aminotransferase concentrations, CD45RO+ cell counts and the liver histologic grade and CD45RO+ cell counts and CD8+ cell counts. These results suggest that CD8+ memory T cells participate in hepatocyte injury in chronic hepatitis C, and that a decrease of CD8+ memory T cells correlates with the decreased liver inflammation with IFN treatment. PMID- 9218826 TI - Antipolysaccharide antibodies in 450 children with otitis media. AB - We have measured antibodies to pneumococcal and Haemophilus polysaccharides in a prospective study of 450 children aged 2-16 years with otitis media requiring grommets (ear tubes). Pneumococcal antibody levels were significantly higher in the 2-6 year (P < 0.004) and 7-10 year (P < 0.04) study groups in comparison with age-matched controls. There was no difference in Haemophilus antibody levels between the study and control group children for the age groups 2-6 years and 11 16 years. Haemophilus antibody levels were significantly lower in the 7-10 year (P < 0.003) group in comparison with age-matched controls. Eighty-eight out of 450 (19.6%) children had pneumococcal antibody levels below the 25th percentile. Nineteen out of 88 (21.6%) children with pneumococcal antibody levels below the 25th centile were test immunized with 23 valent Pneumococcal polysaccharide and unconjugated Haemophilus type b capsular polysaccharide. Of these 19 children (aged 4-11 years), five mounted suboptimal responses to both polysaccharide antigens, whilst one child failed to respond to Haemophilus polysaccharide alone. There was no significant difference in the prevalence of IgG subclass deficiency between the normal responders and poor responders to immunization (P = 0.12). We found no evidence of specific polysaccharide antibody deficiency in the vast majority of the 450 children studied. However, the significance of poor antibody responses to test immunization in a small minority of children with otitis media is unclear. Long-term follow up of these children is required to determine whether poor immunization responses herald the development of frank antibody deficiency. PMID- 9218827 TI - Protective effect of granulocyte colony-stimulating factor (G-CSF) in a granulocytopenic mouse model of Pseudomonas aeruginosa lung infection through enhanced phagocytosis and killing by alveolar macrophages through priming tumour necrosis factor-alpha (TNF-alpha) production. AB - We investigated the effects of G-CSF in a granulocytopenic mouse model of Pseudomonas aeruginosa lung infection. The model was prepared by intratracheal instillation of the bacteria, while granulocytopenia was induced by intraperitoneal injection of 4.0 mg of cyclophosphamide (CPA). There was no difference in the survival rate between G-CSF-treated animals and the normal group, and the number of neutrophils in the blood and lung recovered to normal in the former group. However, the phagocytic and killing activities of neutrophils were lower in G-CSF-treated mice than in controls. Interestingly, the mortality rate increased significantly when anti-TNF-alpha antibody was combined with G CSF, although it was intermediate between CPA alone and CPA-G-CSF-treated mice. However, the improved mortality was not associated with a change in the number of neutrophils in the circulation and lung. Administration of anti-TNF-alpha antibody resulted in a significant suppression of TNF-alpha in bronchoalveolar lavage fluid and of enhanced alveolar macrophage function (phagocytic and bactericidal activity) against P. aeruginosa in G-CSF-treated granulocytopenic mice. We showed also increased TNF-alpha mRNA expression and TNF-alpha production in vitro using G-CSF-pretreated alveolar macrophages compared with control untreated macrophages. Our results are the first evidence to suggest that G-CSF provides a synergistic protective effect against lethal P. aeruginosa lung infection in the granulocytopenic host. This effect is probably due to enhancement of alveolar macrophage function through endogenous TNF-alpha production, in addition to increasing the number of circulating neutrophils. PMID- 9218828 TI - Growth of recombinant Mycobacterium tuberculosis H37Ra in mouse macrophages. AB - Mycobacterium tuberculosis H37Rv and H37Ra were derived from the same parental strain but differ strikingly in their virulence for experimental animals. Transfer of genetic material between these closely related strains resulted in the isolation of a number of recombinant H37Ra clones bearing the in vivo growth promoting ivg locus of H37Rv. The recombinant strain was phagocytosed by murine peritoneal macrophages infected in vivo or in vitro and their intracellular growth rates were compared with the vector control. The intracellular growth of the recombinant was significantly faster than the vector control, but substantially slower than the wild-type H37Rv control, regardless of the method used to infect the macrophages. The slower intracellular growth observed for the recombinant strains was not due to a genetically induced metabolic defect, since they grew in synthetic liquid medium at rates equal to those observed for both H37Rv and H37Ra. Peritoneal macrophage monolayers provide a rapid and convenient assay by which to screen H37Ra recombinants for the presence of putative virulence genes. PMID- 9218829 TI - Elevated plasma levels of IgE in Plasmodium falciparum-primed individuals reflect an increased ratio of IL-4 to interferon-gamma (IFN-gamma)-producing cells. AB - People living in Plasmodium falciparum-endemic areas frequently have elevated levels of total as well as P. falciparum-specific serum IgE. This study aimed at investigating whether the elevated serum IgE levels reflect a shift in the balance between CD4+ T helper 1 (Th1) and T helper 2 (Th2) cells in individuals naturally exposed to the P. falciparum parasite. To investigate the role of Th1 and Th2 cells in the human P. falciparum system we used the ELISPOT assay to determine the ratio of IFN-gamma- and IL-4-producing cells after specific antigen or mitogen activation in vitro. The donors were individuals who had acquired immunity through natural exposure to the parasite. In response to the specific malaria antigens, very few IL-4-producing cells were seen. However, in the response of individual donors to the polyclonal T cell activator, leucoagglutinin (La), the anti-malarial IgE levels in plasma were correlated with an increased ratio of IL-4/IFN-gamma producing cells. Thus, donors with ratios of IL-4/IFN gamma > 1 exhibited mean plasma anti-malarial IgE levels significantly greater than those with ratios < 1. In individuals not living in P. falciparum-endemic areas the ratio of IL-4/IFN-gamma was always < 1. Taken together, our data suggest a shift in the balance between Th1 and Th2 cells in naturally P. falciparum-primed individuals, associated with elevated anti-P. falciparum plasma IgE levels. The role and biological significance of IgE (Th2-type immune response) for protection against P. falciparum and/or pathogenesis of malaria require further study. PMID- 9218830 TI - IL-5 expressed by CD4+ lymphocytes from Echinococcus multilocularis-infected patients. AB - IL-5 is a major factor inducing differentiation of B lymphocytes into immunoglobulin-producing cells as well as a main regulator of eosinophils. Recently, we have shown that peripheral blood mononuclear cells (PBMC) from patients with alveolar echinococcosis (AE) express IL-5 mRNA after stimulation with crude Echinococcus multilocularis (E.m.) antigen. To characterize the observed response in lymphocyte subpopulations, we cultured patients' PBMC in the presence of E.m. crude antigen for 18 h. PBMC were separated from seven patients by fluorescence-activated cell sorting (EPICSorter) into CD4+ and CD8+ subpopulations and from an additional seven patients by magnetic cell sorting (MACS) into CD4+, CD8+ and the CD4+/CD8+ depleted fractions. mRNA was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) for the cytokines IFN gamma, IL-2, IL-3, IL-4, IL-5, IL-6, IL-10, as well as for beta-actin as control. IL-4 and IFN-gamma expression was positive in all of the patients in the stimulated CD4+ subgroup. IL-5 mRNA expression was detected in eight out of 14 CD4+ samples (58%) and not observed in the other subpopulations, or the unstimulated and healthy controls. Co-expression of other Th2 cytokines in the eight patients expressing IL-5 mRNA was found in five patients for IL-3 and in seven for IL-10. Expression of IL-5 and both Th2 cytokines (IL-3 and IL-10) was only observed in patients judged as critically ill. Out of the six patients who were regarded as cured after radical operation or as stabilized with or without chemotherapy, only two expressed IL-5. Out of those eight patients considered as critically ill, six expressed IL-5 mRNA and five of these co-expressed IL-3 and IL-10. Thus, we conclude that specific antigenic challenge of PBMC from patients with active or previous AE induces an IL-5 response of CD4+ lymphocytes. The expression of Th2-type interleukin mRNA is significantly more frequent in patients clinically judged as progressive. Furthermore, IgE was elevated only in patients regarded as critically ill (six out of eight). In none of the patients were eosinophils elevated. These data support a Th2-type immune response in patients with chronic E. multilocularis infection. PMID- 9218831 TI - Hyperexpression of transporter in antigen processing-1 (TAP-1) in thyroid glands affected by autoimmunity: a contributing factor to the breach of tolerance to thyroid antigens? AB - According to the 'aberrant HLA expression' hypothesis, endocrine autoimmunity is driven by presentation of self antigens by target cells over-expressing HLA molecules. In autoimmune thyroid diseases (AITD), thyroid follicular cells (thyrocytes) over-express HLA class I and HLA class II molecules. Since efficient presentation of endogenous peptides via class I requires transporters that translocate endogenous peptides from the cytoplasm to the endoplasmic reticulum, i.e. transporters associated with antigen processing (TAP) -1 and -2, the capability of thyrocytes to express TAP and whether TAP is hyperexpressed in AITD glands are issues relevant to the above hypothesis. Results from immunofluorescence and Northern blotting studies on primary thyrocyte cultures and on a thyroid cell line demonstrate that thyrocytes express constitutively TAP 1 at a low level, and that this expression is readily induced by interferon-gamma (IFN-gamma) and to a lesser extent by IFN-alpha. In AITD, but not in non autoimmune glands, thyrocytes hyperexpress TAP-1, as demonstrated by both immunohistopathology and flow cytometry. The cytokine pattern does not bear, as assessed by reverse transcriptase-polymerase chain reaction (RT-PCR), a clear relationship with TAP-1 expression. These results have broad implications and suggest that the core concept of the 'aberrant HLA expression' hypothesis of endocrine autoimmunity could be incorporated in the currently prevailing view of 'autoimmunity by breach of peripheral tolerance'. PMID- 9218832 TI - Up-regulated beta1-integrin expression in autoimmune thyroid disorders. AB - Lymphocytic infiltration of the thyroid gland in autoimmune thyroid disorders requires, as a first step, their attachment to endothelial cells (EC) and, subsequently, interaction with thyrocytes and extracellular matrix proteins. Recent studies have focused on the pathophysiologic role of beta1-integrins as adhesion receptors for extracellular matrix proteins and as cell-to-cell adhesion receptors. In this study, we examine by flow cytometry and immunohistochemical techniques the differences in expression of beta1-integrins in thyrocytes and EC between normal thyroids and thyroid glands from patients with Graves' disease (GD) and Hashimoto's thyroiditis (HT). Remarkably, we found an up-regulated de novo expression of very late antigen (VLA)-alpha6 subunit in thyrocytes in close proximity to lymphocyte infiltrates in GD and HT thyroid glands, with no reactivity in control thyroids. Moreover, interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF-alpha) and IL-1beta produced a significant enhancement of VLA-alpha6 expression in vitro in thyrocytes in culture. In addition, an up regulated expression of VLA-alpha5 and beta1 subunits was found in thyrocytes from GD and HT glands, specifically in those areas more severely inflamed. VLA alpha2 was basally expressed in middle size and large vessels in control glands, with an increased expression in vessels of all sizes in HT and GD glands. Dendritic cells in thyroid lymphoid follicles were also positive for VLA-beta1, alpha2 and alpha6 subunits. These results indicate the existence of an up regulatory process in the expression of beta1-integrins, particularly the alpha6 subunit, in several cell types from inflamed GD and HT thyroid glands, suggesting that these integrins could play a relevant role in localizing and perpetuating the autoimmune response in the thyroid gland in autoimmune thyroid disorders. PMID- 9218833 TI - Streptozotocin-induced diabetes in mice lacking alphabeta T cells. AB - Multiple low-dose streptozotocin (MD-STZ) is widely used for the experimental induction of diabetes, but, as non-obese diabetic (NOD)-scid/scid mice have been found to display enhanced susceptibility to MD-STZ, whether or not the model is genuinely autoimmune and T cell-mediated has been unclear. Mice bearing a targeted mutation of the T cell receptor (TCR) alpha-chain were therefore used to assess whether TCR alphabeta+ cells are involved in the diabetogenic effects of MD-STZ injections. Young NOD mice lacking TCR alphabeta cells, when given five daily injections of 40 mg/kg STZ, developed diabetes at low frequency (2/12), despite the widespread destruction of pancreatic islet cells. By comparison, most normal control mice became hyperglycaemic (12/23). We conclude that whilst much of the tissue destruction observed in this model is due to the direct toxic effect of STZ, a significant amount is also due to the action of TCR alphabeta cells tipping the balance between tolerable and clinically damaging action on islet cells. PMID- 9218834 TI - Stimulation of peripheral blood lymphocytes with Campylobacter jejuni generates a gammadelta T cell response in patients with Guillain-Barresyndrome. AB - In three patients whose Guillain-Barre syndrome (GBS) was preceded by gastrointestinal infection due to Campylobacter jejuni, gammadelta T cells were generated from peripheral blood in response to in vitro stimulation with C. jejuni. In one of the patients, where a diagnostic sural nerve biopsy was performed, gammadelta T cells were also isolated following culture of the nerve tissue. Studies with healthy volunteers and C. jejuni gastroenteritis patients also showed preferential enrichment for gammadelta T cells in peripheral blood cells stimulated with C. jejuni, although the response was significantly lower than that seen in GBS patients. In two out of three GBS patients and all of the controls, gammadelta T cell receptor (TCR) gene usage was shown to be Vgamma9/Vdelta2+. In the GBS patient where nerve-infiltrating gammadelta T cells were isolated, these and C. jejuni-specific peripheral blood cells had similar TCR gene usage, predominantly consisting of Vgamma5/Vdelta1+ cells. Sequencing the Vdelta1 products from nerve and peripheral blood showed similarities in CDR3 length, but the single Vdelta1 sequence obtained from nerve was not identified in peripheral blood. These results suggest that the generation of gammadelta T cells is part of a normal immune response to C. jejuni, which, in patients with GBS, may contribute to the pathogenesis of their inflammatory neuropathy. PMID- 9218835 TI - Intracerebral injection of myelin basic protein (MBP) induces inflammation in brain and causes paraplegia in MBP-sensitized B6 mice. AB - Brain inflammation and paraplegia can be induced by an additional intraperitoneal (i.p.) and intracerebral (i.c.) restimulation in B6 mice after standard immunization with MBP in Freund's complete adjuvant (FCA) and Bordetella pertussis coadjuvant. Only the combination of i.p. MBP/FCA and i.c. MBP injection could induce clinical paraplegia; either one alone was not effective. Clinical symptoms would develop 2 days after the i.c. injection. The induction of paraplegia was MBP-specific, as irrelevant bovine serum albumin with the same protocol could not induce it. The i.p. restimulation was requisite and needed the MBP in FCA, as MBP in PBS was ineffective. Histopathological observation manifested cellular infiltration by leucocytes in perivascular spaces and cerebral cortex. Neutrophils were prominent at 12 h after i.c. injection, then were replaced by mononuclear cells 24 h later. There were dynamic changes in cell number and immunophenotype of VLA-4+ expression in cervical lymph node cells after i.c. injection. The cells derived from cervical lymph nodes had higher MBP stimulated proliferation than that of distal lymph nodes. This additional i.p. and i.c. stimulation provides a new manipulation to study brain inflammation. PMID- 9218836 TI - Mechanisms of T cell-induced glomerular injury in anti-glomerular basement membrane (GBM) glomerulonephritis in rats. AB - The effector mechanisms of T cell-dependent acute glomerular injury were studied in autologous phase anti-GBM glomerulonephritis (GN) in rats. Acute proliferative GN was induced in sensitized rats by a subnephritogenic dose of sheep anti-rat GBM antibody. Injury was manifested by proteinuria and glomerular leucocyte infiltration composed predominantly of macrophages but also CD4+ and CD8+ T cells. T cell depletion, using an anti-CD5 MoAb, demonstrated that glomerular leucocyte infiltration and proteinuria were T cell-dependent. Inhibition of T helper cell function using an anti-CD4 MoAb prevented proteinuria and glomerular macrophage and CD4+ T cell influx, but not accumulation of CD8+ T cells. Depletion of CD8+ T cells also prevented proteinuria and the influx of macrophages and CD8+ T cells, but not accumulation of CD4+ T cells. Macrophage depletion, using micro-encapsulated clodronate, prevented proteinuria and glomerular macrophage infiltration, but not the accumulation of CD4+ or CD8+ T cells, indicating that macrophages are the common cellular effectors for both CD4 and CD8 T cell-dependent injury. Evidence for cytotoxic mechanisms of injury (increased numbers of apoptotic cells or accumulation of natural killer (NK) cells in glomeruli) could not be demonstrated. These data suggest that acute glomerular injury in anti-GBM GN is the result of macrophage recruitment, which is dependent on both CD4 and CD8 T cells, and that direct T cell-mediated injury (cellular cytotoxicity) is not involved. PMID- 9218837 TI - Gene expression of CC chemokines in experimental crescentic glomerulonephritis (CGN). AB - CGN is a rapidly progressive glomerular disease. Monocytes/macrophages are frequently observed in glomeruli in cases of CGN and they are considered to play a crucial role in the pathogenesis of this disease. We described previously the glomerular expression of monocyte chemoattractant protein-1 (MCP-1), which is a potent chemoattractant for monocytes and a member of CC chemokine family, in an experimental model of CGN. In the present study we investigated the expression of mRNAs for other CC chemokines, namely, MCP-3, macrophage inflammatory protein 1alpha (MIP-1alpha), MIP-1beta, RANTES and TCA3, all of which are chemotactic for monocytes, in the CGN model. First, we established a reverse transcriptase polymerase chain reaction (RT-PCR) method by which mRNA for each of the CC chemokines could be amplified separately, and then we measured the levels of the expression of mRNAs for the chemokines in diseased glomeruli at several time points after induction of CGN. The mRNAs for all CC chemokines examined were expressed in glomeruli of rats with CGN. Moreover, induction of the gene expression of MIP-1alpha and MIP-1beta seemed to occur earlier than that of the others. CC chemokines may contribute to the recruitment and activation of monocytes in CGN, and each individual CC chemokine may play an overlapping but distinct role in the pathogenesis of this disease. PMID- 9218838 TI - Genetic differences in immune reactivity to mercuric chloride (HgCl2): immunosuppression of H-2d mice is mediated by interferon-gamma (IFN-gamma). AB - Upon treatment with HgCl2, H-2s mice, such as B10.S, develop an activation of B lymphocytes that depends, at least partially, on activation of T helper type 2 (Th2) cells and results in increased serum levels of IgG1 and IgE, appearance of IgG autoantibodies, and development of immune glomerulonephritis and vasculitis. Results of previous studies and of experiments presented here indicate that the B cell activation and systemic autoimmune disease fail to develop in MHC-congenic B10.D2 (H-2d) and B10.BR (H-2k) mice treated with HgCl2, although B10.D2 T cells showed signs of activation by and specificity for HgCl2 comparable to those seen in strain B10.S. Here, we report that following HgCl2 injections the antibody response to sheep erythrocytes is normal in B10.S, but suppressed in B10.D2 mice. This suppression was prevented by MoAb to mouse IFN-gamma. Conversely, treatment of B10.D2 mice with murine recombinant IFN-gamma (rIFN-gamma) was able to reproduce the immunosuppression seen after HgCl2 treatment. In B10.S mice, it took administration of both rIFN-gamma and HgCl2 to suppress the anti-sheep erythrocyte response. Although rIFN-gamma diminished the increase in IgE serum levels of HgCl2-treated B10.S mice, it failed to prevent their autoantibody production and immune glomerulonephritis. These findings further strengthen the concept that B10.S mice react to HgCl2 by preferential activation of their Th2 cells producing IL-4, whereas B10.D2 mice react to HgCl2 by preferential activation of their Th1 cells, which produce IFN-gamma and thus suppress antibody responses. PMID- 9218839 TI - Humoral response against heat shock proteins and other mycobacterial antigens after intravesical treatment with bacille Calmette-Guerin (BCG) in patients with superficial bladder cancer. AB - Few studies have analysed the antibody response during intravesical BCG immunotherapy for superficial bladder cancer. We have examined the evolution in serum antibody response against several heat shock proteins (hsp), including the recombinant mycobacterial hsp65 and the native protein P64 from BCG, GroEL from Escherichia coli (hsp60 family), recombinant mycobacterial hsp70 and the E. coli DnaK (hsp70 family), against purified protein derivative of tuberculin (PPD) and the AG85 complex of Mycobacterium bovis BCG, as well as against tetanus toxoid in 42 patients with a superficial bladder tumour, 28 treated with six intravesical BCG instillations and 14 patients used as controls. We also analysed the lymphoproliferative response of peripheral blood mononuclear cells against PPD in this population. Data of antibody responses at 6 weeks post BCG were available in all 28 patients, and at 4 month follow up in 17 patients. All patients who demonstrated a significant increase in IgG antibodies against PPD at 4 months follow up had a significant increase already at 6 weeks of follow up. In contrast, IgG antibodies against hsp increased significantly from 6 weeks to 4 months post-treatment. A significant increase in IgG antibodies against PPD, hsp65, P64, GroEL, and hsp70 at 4 months follow up was observed in 10/17, 8/17, 10/17, 4/17 and 8/17 patients. Native P64 protein elicited a higher antibody response than recombinant mycobacterial hsp65. No increase in antibody response was observed against Dnak from E. coli, against AG85 or tetanus toxoid after BCG therapy. An increase in IgG antibodies against P64 at 4 months follow up compared with pretreatment values was found to be a significant predictor of tumour recurrence (P<0.01). Further studies with a larger number of patients are needed to confirm the value of the antibody response against P64 as a clinical independent prognostic factor. PMID- 9218840 TI - Isolation of human anti-c-erbB-2 Fabs from a lymph node-derived phage display library. AB - An immunoglobulin phage display library constructed from a tumour-associated pericolic lymph node was panned against the extracellular domain of the oncoprotein c-erbB-2. Sixteen independent clones were confirmed as positive binders based on ELISA analysis of soluble Fabs. Nucleotide sequencing demonstrated that the V(H) region of 12 clones belonged to four different V gene families, and the clones demonstrated varying degrees of somatic mutation compared with germ-line sequences. Fab fragments were examined for cross reactivity by ELISA and shown to be negative against a panel of irrelevant self and non-self antigens, including bovine serum albumin (BSA), mouse immunoglobulin, tetanus toxoid, heregulin-PE40-FLAG and insulin. Reactivity of Fabs in vitro was verified by immunocytochemistry, which showed binding to the c erbB-2 over-expressing breast cancer cell line SKBR3 but not to the low expressing cell line MDA-MB-231. We conclude that a single lymph node library of moderate diversity (2 x 10(7) kappa light chain and gamma heavy chain clones), when derived from an individual whose colorectal tumour over-expressed c-erbB-2, can be successfully panned to isolate a number of unique Fabs specific for this antigen. The nature of the anti-c-erbB-2 Fabs recovered from this library suggests that they may have resulted from a humoral immune response in the individual, and that in vivo antibody responses to tumour-associated antigens may be exploited in vitro for the production of tumour-specific recombinant antibodies. PMID- 9218841 TI - Evidence for genetic heterogeneity in inflammatory bowel disease (IBD); HLA genes in the predisposition to suffer from ulcerative colitis (UC) and Crohn's disease (CD). AB - Family and epidemiological studies support a genetic susceptibility to UC and CD. Conflicting reports regarding associations between UC and HLA-DR2 and between CD and various HLA alleles have been published. The aim of this study was to determine whether molecularly defined HLA-DR genes are associated with these diseases in a Dutch group of patients. Fifty-nine unrelated Dutch UC patients and 89 CD patients were typed using DNA-based methods. A total of 2400 healthy local blood donors served as controls. The phenotype frequency of the HLA-DRB1*15 allele was increased in UC patients compared with controls (42% versus 26% in controls; P = 0.006; odds ratio (OR) = 2.1), and was predominantly found in female patients (53% versus 24%; P = 0.001; OR = 3.5). The DRB1*15 allele was increased in UC patients having a positive family history (P = 0.01; OR = 5.8). Among the 16 patients who showed an increase in extent of disease during follow up, 10 were DRB1*15+ (P = 0.002; OR = 4.8). The frequency of the DRB1*13 allele was decreased in patients with UC (15% versus 28% in controls; P = 0.04; OR = 0.5). In CD, no association was observed between disease or particular clinical subgroups and any allele tested. The present study provides additional evidence for the genetic association between UC and HLA-DRB1*15, and supports recent findings that the susceptibility gene(s) for CD is not located in the HLA class II region. PMID- 9218842 TI - Induction and suppression of anti-antibodies to syngeneic T cell-binding antibodies in mice. AB - Considerable effort is being invested in antibody gene technologies for production of species-adapted anti-T cell antibodies which can overcome formation of neutralizing anti-antibodies (anti-Ab). By establishing a mouse model for the generation of syngeneic anti-T cell MoAb, we addressed the question of whether ideally species-adapted T cell-binding antibodies can prime mice to produce anti Ab. Two anti-Thy-1.2 MoAbs of IgG2a (MmTC) or IgM (MmTC-IgM) isotype were generated in congenic C57B1/6-Thy-1.1 mice. Three injections of MmTC in C57B1/6 Thy-1.2 or (C57B1/6-Thy-1.2xC57B1/6-Thy-1.1)F1 hybrids where they act as syngeneic anti-T cell MoAbs induced low anti-Ab. When MmTC was injected as Igh mismatched antibody in CBA/J mice, high titre anti-MmTC antibodies were measured and fully mismatched skin allografts rejected within 26 days. MmTC injected twice weekly in syngeneic C57B1/6 mice induced low anti-MmTC and prolonged graft survival up to day 117. In contrast, retreatment induced anti-MmTC with graft rejection within 32 days. Thus, upon retreatment, the immunosuppressive effects of MmTC were inhibited to a similar extent as that seen after antibody treatment in Igh-mismatched mice. However, a recently analysed immunological principle to suppress anti-Ab against T cell binding allo- or xenoantibodies by preinjection of T cell-depleting antibody with species differences in heavy chains also suppressed syngeneic anti-Ab after MmTC retreatment. This effect was accompanied by prolonged graft survival. Thus, our data indicate that inhibitory anti-Ab must be considered in humans even when treated with fully species-adapted anti-T cell MoAb, but may be suppressed by preinjection of a Fc region-mismatched anti-T cell antibody. PMID- 9218843 TI - Rescue by cytokines of apoptotic cell death induced by IL-2 deprivation of human antigen-specific T cell clones. AB - The control of cell survival and cell death is of central importance in tissues with high cell turnover such as the lymphoid system. We have examined the effect of cytokines on IL-2 deprivation-induced apoptosis of human antigen-specific T helper clones with different cytokine production profiles. We found that IL-2, interferon-alpha (IFN-alpha), and IFN-beta inhibited IL-2 deprivation apoptosis in Th0, Th1, and Th2 clones. We also found that IL-2 protects T cell clones from IL-2 deprivation apoptosis accompanying active proliferation and enhanced expression of P53, Rb and Bcl-xL proteins. In contrast, IFN-alpha/beta rescued T cell clones from apoptosis without active proliferation, and expression of apoptosis-associated proteins tested so far was unaffected. This may be due to the fact that T cells treated with IL-2 contained those located in S + G2/M phases of the cell cycle, whereas the vast majority of T cells treated with IFN alpha/beta were located in G0/G1 phase. IFN-alpha/beta specifically induced tyrosine phosphorylation and translocation into nucleus of signal transducers and activators of transcription (STAT) 2 protein in the T cell clones. In addition, over-expression of STAT2 by transfection of the cDNA prevented apoptosis of the T cell clones. Our present study shows that IFN-alpha and -beta mediate anti apoptotic effect through other pathways than that of IL-2 in growth factor deprivation apoptosis. PMID- 9218844 TI - The normally expressed kappa immunoglobulin light chain gene repertoire and somatic mutations studied by single-sided specific polymerase chain reaction (PCR); frequent occurrence of features often assigned to autoimmunity. AB - The expressed human kappa light chain gene repertoire utilized by healthy individuals was studied by two different single-sided specific PCR techniques to avoid bias for certain V genes. A total of 103 rearranged kappa sequences from peripheral blood mononuclear cells from healthy individuals were cloned from cDNA and assigned to the Vkappa and Jkappa germ-line genes with the closest overall homology. The use of cDNA rather than genomic DNA focused the analysis on activated B cells rich in mRNA. Accordingly, the sequences represented the applied repertoire and almost all were somatically mutated. V genes from the Jkappa-proximal duplication unit of the kappa locus were almost exclusively used. A total of 65% of the sequences could be assigned to four or five genes: A27 (humkv325), L6 (Vg), L2 (humkv328), and A3 and/or A19. N additions and P nucleotides were quite common and found in 32% and 21% of the sequences, respectively. Extended CDR3s more than nine residues in length were found in 18% of the sequences, and in 71% of cases this was due to insertion of an extra proline residue. This proline was usually explained from the germ-line sequences involved. These results are in good agreement with those of previous repertoire studies using potentially V-gene-biased techniques. Thus, it is clear that restricted V-gene usage, common N and P additions, and extended CDR3 regions are normal features and not, as has been claimed, characteristics of pathological autoantibodies. PMID- 9218845 TI - Surface-related triggering of the neutrophil respiratory burst. Characterization of the response induced by IgG adsorbed to hydrophilic and hydrophobic glass surfaces. AB - Hydrophilic and hydrophobic glass surfaces precoated with human albumin, fibrinogen, or IgG were investigated with respect to their ability to activate the neutrophil NADPH-oxidase. We found that IgG-coated surfaces induced a substantial and prolonged neutrophil production of reactive oxygen species (ROS). When a hydrophilic surface was used to support protein binding, a somewhat lower neutrophil response (around 35%) was obtained, compared with the response induced by IgG on a hydrophobic surface. The production of ROS was completely eliminated when cytochalasin B was added to the measuring system, suggesting the involvement of the cell cytoskeleton in the activation process. The relation between the intra- and extracellular generation of ROS was further assessed, and we found that most of the ROS produced were released from the cells, in agreement with a model in which the activating surfaces induce a 'frustrated' phagocytic response. Serum totally inhibited 'frustrated' phagocytosis provided that the IgG molecules were sticking to a hydrophilic surface. PMID- 9218846 TI - Binding and internalization of human IgG by living cultured endothelial cells. AB - Interactions between circulating IgG and endothelial cells (EC) in humans have been described only in conditions associated with pathologic immunoglobulins and/or activated or damaged EC. In this study we provide evidence that normal human IgG includes one/some antibody species that bind to and are internalized by living EC in culture. This novel function of EC and natural autoantibodies is of potential importance for the understanding of physiologic interactions between vessels and the immune system and for the clarification of pathogenesis of vasculitis and mechanisms of action of pooled IgG used in the therapy of such conditions. PMID- 9218847 TI - Complement coating of erythrocytes is reduced following their interaction with neutrophils in vitro without loss of complement receptor 1 (CR1). AB - We have investigated the interaction of complement-coated erythrocytes (E) with neutrophils, in vitro, to determine the effects of erythrocyte CR1. Complement coating in vitro, to mimic immune complex uptake, caused a reduction in E-CR1, but subsequent interaction with neutrophils effected a removal of E-C3b and a return of E-CR1 to levels approximating that of uncoated erythrocytes. These data indicate that C3b, associated with immune complexes, may bind to E-CR1 in a reversible manner and that subsequent interaction with phagocytes need not necessarily result in cleavage (and loss of) E-CR1. Further, we conclude that E CRI can apparently be reduced by epitope masking. PMID- 9218848 TI - [New information on a classic antidepressive agent]. PMID- 9218849 TI - Injection of externally generated ions into an increasing trapping field of a quadrupole ion trap mass spectrometer. AB - Trapping ions injected into a quadrupole ion trap (QIT) by increasing the trapping r.f. voltage on a ring electrode is an effective and widely recognized method of interfacing an ion trap with pulsed ion sources such as matrix-assisted laser desorption/ionization (MALDI). In this paper, the problem of mass discrimination during the injection and trapping of ions by the increasing r.f. field was studied both experimentally and by numerical simulation using SIMION software. For a MALDI/QIT interface design with a remote external ion source described here, experiments with polyethylene glycol (PEG 1000 and PEG 1500) showed little mass discrimination for trapping ions in a wide mass range (500 2000 Dn) for a broad range of experimental conditions, which include kinetic energies of 5-40 eV for the injected ions and an r.f. voltage of 400-4000 Vo-p amplitude ramped at a rate of 30-140 Vo-p mus-1. In the numerical simulation, complex and sharp dependences of the trapping efficiency on the phase of the r.f. voltage and initial kinetic energy of ions were observed. However, after averaging over the r.f. phase and over a reasonable range of kinetic energy, the simulation resulted in relatively constant and high values for the trapping efficiency (normally 0.2-0.3) for any mass and kinetic energy considered, which are consistent with the weak sensitivity to injection parameters observed in the experiment. A simple model for the qualitative description of ion injection and trapping is suggested that relies on phase interaction of injected ions with the r.f. field rather than on collisions with the buffer gas molecules to decrease the ion kinetic energy. PMID- 9218851 TI - Current literature in mass spectrometry. PMID- 9218850 TI - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry as a tool to probe the reactions of trans-Hex-2-enal with proteins. PMID- 9218852 TI - How should we treat hepatitis C in 1997? PMID- 9218853 TI - Failure of ketoprofen and interferon combination therapy to improve interferon resistant chronic hepatitis C. AB - Preliminary reports suggest that patients with interferon (IFN)-resistant chronic hepatitis C respond better to a combination of IFN-alpha and nonsteroidal anti inflammatory drugs than to IFN alone. The efficacy of IFN combined with ketoprofen in the treatment of patients with IFN-resistant chronic hepatitis C was evaluated. Seventeen patients, nonresponsive after at least six months of treatment with IFN-alpha 2b and subsequently treated with the combination of IFN alpha 2b plus ketoprofen for four months, were studied. Serum aminotransferases (alanine aminotransferase [ALT] and aspartate aminotransferase [AST]) and serum hepatitis C virus (HCV) RNA were analyzed before and throughout treatment. No patient normalized serum aminotransferases after combination therapy. There were no significant differences in mean serum ALT and AST levels before and after ketoprofen intervention. Serum HCV RNA became undetectable after treatment in only one patient, but was detectable again three months after treatment cessation. These results provide no convincing evidence that the combination of IFN-alpha 2b with ketoprofen improves the response to IFN in patients nonresponsive to IFN alone. PMID- 9218854 TI - Helicobacter pylori: primary susceptibility to clarithromycin in vitro in Nova Scotia. AB - Resistance to antimicrobial agents is a major determinant of the efficacy of regimens to eradicate Helicobacter pylori. Clarithromycin (CLA) has become one of the most commonly used antibiotics for treatment of H pylori infection. In this study, the rate of primary resistance to CLA in H pylori isolated from patients was determined. One hundred sixty-two strains were recovered from patients before treatment. Strains were grown and inoculated onto Mueller-Hinton agar with 7% sheep blood. CLA epsilometer gradient agar diffusion test (E test) strips were used to test for susceptibility. Appropriate control organisms were tested to validate the assay. Plates were incubated at 37 degrees C in a microaerophilic atmosphere for up to five days. E test results were easy to interpret. Strains were considered resistant if the minimum inhibitory concentration (MIC) was 2 micrograms/mL or greater. Three strains were resistant (two strains with MIC 8 micrograms/mL and one strain with MIC 12 micrograms/mL) and 159 strains were sensitive (MICs ranged from less than 0.016 to 0.38 micrograms/mL). Ninety per cent of the strains had MICs of 0.023 micrograms/mL. Primary resistance was 1.8%. These susceptibility data support the use of CLA for the treatment of H pylori in the Nova Scotia population. PMID- 9218855 TI - Atypical perinuclear antineutrophil cytoplasmic antibodies after colectomy in inflammatory bowel disease. AB - A typical perinuclear antineutrophil cytoplasmic antibodies (p-ANCA) have been detected in most patients with ulcerative colitis and primary sclerosing cholangitis. Persistent atypical p-ANCA have been observed in ulcerative colitis patients with a prior proctocolectomy, especially with pouchitis, suggesting that this serological marker might be predictive of subsequent development of chronic or refractory pouchitis. This study prospectively evaluated this serological marker in 24 consecutive patients with inflammatory bowel disease and prior colectomies (12 with a clinical diagnosis of ulcerative colitis and 12 with a clinical diagnosis of Crohn's disease involving the colon). Of these, 14 were positive, including 11 with extensive ulcerative colitis and three with Crohn's disease. Although two of three ulcerative colitis patients with pouchitis were positive, eight of eight ulcerative colitis patients having a pelvic pouch with no] pouchitis were also positive, as was a patient who elected to have an end ileostomy (Brooke's ileostomy). Two patients had abnormal liver chemistry tests. Both had end-stage primary sclerosing cholangitis treated with liver transplantation and were positive for this serological marker. Although atypical p-ANCA may be a marker of persistent inflammation in pelvic pouch patients, a positive test result should not be used for prognosis or as a decision-making parameter for pelvic pouch procedures. PMID- 9218856 TI - Duodenal ulcer and Helicobacter pylori infection at high altitude: experience from southern Saudi Arabia. AB - OBJECTIVE: To study the clinical presentation, endoscopic features and prevalence of Helicobacter pylori in duodenal ulcer (DU) patients in southern Saudi Arabia, located 3150 m above sea level, and to compare results with those from low altitude regions of the Kingdom. METHODS: Prospective study of patients with proven DU referred for upper gastrointestinal endoscopy at Asir Central Hospital, Abha, southern Saudi Arabia over an 18-month period. RESULTS: Of 126 patients with proven DU, 72% were men and mean age was 40.4 years (range 18 to 68). Twenty eight per cent were smokers and only 5% used nonsteroidal anti-inflammatory drugs. Thirty-eight patients (30%) presented with hematemesis or melena, and the majority had a single ulcer. Nineteen per cent of patients with dyspepsia had DU and 96% had H pylori. These results are comparable with those reported from the low altitude, warmer regions of Saudi Arabia. CONCLUSIONS: Age of patients and the male:female ratio were similar to those in developing countries. The frequency of smoking is lower than in western countries and no patient in this report consumed alcohol. High altitude did not affect the prevalence of DU or the frequency of H pylori because the results were comparable with those from the low altitude areas of the Kingdom of Saudi Arabia and other lowland developing countries. Although great socioeconomic changes have increased the incidence of heart disease, the patterns of DU and H pylori infection assume those in developing nations. PMID- 9218857 TI - National survey of radionuclide gastric emptying studies. AB - A survey was mailed to all institutions in Canada licensed to use radiopharmaceuticals. Questions addressed meal type; mode of preparation; and means, ranges and SD of emptying times. Seventy-eight per cent of 222 facilities responded, including all 55 teaching centres. Eighty-five per cent of teaching and 56% of nonteaching centres perform solid phase gastric emptying studies (GES). The majority use 99mTc sulphur colloid (Tc-SC) added to eggs before cooking as the standard meal. Twenty-five per cent of teaching and 21% of nonteaching centres perform liquid phase GES. Most use a watery solution of 111Indiethylenetriamine pentaacetic acid. Gastric emptying for solid phase GES, expressed as time for 50% emptying (mean t1/2), varied from 42 to 105 mins for centres using the Tc-SC egg meal. Twenty-eight per cent of teaching centres used +/- 2 SD to define their normal range, 26% used +/- 1 SD, 6% used +/- 1.5 SD, and 40% did not know the number of SD used. Twenty per cent of nonteaching centres used +/- 2 SD, 12% used +/- 1 SD and 68% did not know how many SD were used. For liquid phase GES, mean t1/2 varied from 20 to 60 mins. Eighteen per cent of centres used healthy volunteers to establish or validate normal ranges. There is substantial variability among the normal ranges for radionuclide solid and liquid phase GES in both teaching and nonteaching centres across Canada. A minority of facilities have established or validated their own normal ranges in healthy volunteers. There is a need for a more standardized protocol and range of normal, with internal validation by each institution. PMID- 9218859 TI - Use of oral sodium phosphate colonic lavage solution by Canadian colonoscopists: pitfalls and complications. AB - Oral sodium phosphate (NaP) has become an attractive alternative to polyethylene glycol (PEG) for colonic cleansing before colonoscopy, but it potentially has greater complications. This study surveyed members of the Canadian Association of Gastroenterology (CAG) to determine how these colonic lavage agents are used and what complications have been encountered. The Dillman survey technique produced responses from 67% of the 400 members who perform colonoscopy. For the larger out patient group, respondents used NaP more frequently than PEG (46% versus 35%, respectively, P < 0.015). Respondents used NaP and PEG with similar frequencies for the in-patient group (44% versus 43%). Of respondents using NaP, 45% reported excluding its use in patients with renal failure, 30% with heart disease, 13% with incomplete bowel obstruction and 9% with extreme age. Symptoms suggestive of hypovolemia were reported in 9% of those using NaP compared with 3% using PEG (P < 0.02). Three patients receiving NaP developed acute renal failure. A greater proportion of those using NaP had small unexplained aphthous ulcers (16%) and excessive luminal bubbling (24%) compared with PEG users (3%, P < 0.00001 and 14%, P < 0.03, respectively). These data demonstrate that members of CAG use NaP more frequently than PEG as the colonic lavage solution before colonoscopy. A greater number reported complications with NaP versus PEG, and a significant proportion of the respondents appeared to be unaware of the potential for these complications in specific clinical circumstances. PMID- 9218858 TI - Cost effectiveness of alternative Helicobacter pylori eradication strategies in the management of duodenal ulcer. AB - Published data and techniques for decision analysis were used to construct a model to estimate the cost effectiveness of nine alternative strategies for the management of patients diagnosed with uncomplicated duodenal ulcer. Two strategies of intermittent therapy with either ranitidine or omeprazole, one strategy of continuous maintenance treatment with ranitidine, and six strategies for ulcer healing and eradication of Helicobacter pylori infection were considered. Healing time curves were estimated by using published data, allowing for estimation of expected time for acute healing episodes. The expected number of weeks to heal per patient, in a one-year period, was estimated by combining healing time data with probability of ulcer recurrence. It was found that patients that underwent any of the six H pylori eradication regimens had fewer days with ulcer per year than those who underwent maintenance or intermittent ranitidine. Four eradication regimens had lower costs and better outcomes than ranitidine therapy. In comparing H pylori strategies, the two strategies of omeprazole plus one antibiotic (either amoxicillin or clarithromycin) are most costly than omeprazole plus two antibiotics (specifically amoxicillin and metronidazole or clarithromycin and metronidazole) and result in similar outcomes. Although omeprazole-based eradication regimens are more costly than ranitidine bismuth triple therapy, they are associated with fewer recurrences of ulcer and days of symptoms. A limitation of the analysis is that it did not incorporate issues of compliance and metronidazole resistance; however, the former concern may be less of an issue as H pylori regimens become simpler and shorter in duration. PMID- 9218860 TI - Use of octreotide in the acute management of bleeding esophageal varices. AB - Acute hemorrhage from esophageal varices is a medical emergency; despite early diagnosis and treatment the associated hospital mortality remains high. The clinical research summarized in this paper shows that octreotide has a beneficial effect on portal hemodynamics in cirrhotic patients. In randomized controlled trials octreotide has been effective in halting initial hemorrhage and in preventing reoccurrence of bleeding. Somatostatin and octreotide appear to be equivalent in terms of therapeutic efficacy but octreotide is the less expensive option. For suspected variceal bleeding an octreotide infusion should be initiated immediately. To prevent further bleeding the drug should be continued for two to five days after endoscopic variceal ligation. PMID- 9218861 TI - Smoking and diseases of the gastrointestinal system: an epidemiological review with special reference to sex differences. AB - Smoking increases the risk of peptic ulcer disease and death from it. Smoking delays peptic ulcer healing, with or without treatment, and increases the risk of recurrence after healing. The effects of smoking on this disease are similar and equally pervasive in women and men. There is growing evidence that cigarette smoking is a risk factor for Crohn's disease (CD) in both women and men. However, women smokers appear to be at particular risk for this disease. In studies that examined this risk separately in women and men. At each level of smoking the excess risk in women smokers compared with nonsmokers clearly exceeded the excess risk in men smokers compared with nonsmokers. Smoking also appears to adversely affect the clinical course of CD in both women and men, but more so in women. The possible interaction between smoking and oral contraceptives with regard to the risk of CD deserves further study. There is growing evidence that current smoking protects against ulcerative colitis in both men and women. Although there is some evidence that smoking is a risk factor for gallstones, particularly in women, evidence to support a causal relationship is inadequate. Further studies, controlling for alcohol consumption in the analyses, are needed. Smoking does not appear to be a risk factor for cirrhosis of the liver. PMID- 9218863 TI - Bacterial glycosulphatases and sulphomucin degradation. AB - The presence of a high bacterial population in a region of the gastrointestinal tract is usually associated with the secretion of sulphomucins into the mucus gel covering that region. The term 'sulphomucin' is a histochemical description of the staining properties of mucin. At present this term can only be qualitatively related to the percentage of sulphate in the mucin molecule, which makes the term difficult to use in a biochemical and functional sense. Sulphomucins are thought to carry out the normal functions attributed to mucins; in addition, heavy sulphation rate-limits the degradation of mucins by bacterial mucin-degrading glycosidases. A number of mucin-specific glycosulphatases have been reported in bacteria, although only two such enzymes have been purified. These enzymes remove part of the sulphate content from sulphomucins and make them more susceptible to further enzymic degradation. The variety of chain locations and sugar attachment sites of sulphate esters on the mucin oligosaccharides, taken together with the data on the enzymes, suggest there will be a spectrum of bacterial glycosulphatases, with different properties, cellular locations and substrate specificities. Bacterial glycosulphatases have the potential to modify sulphated glycoconjugates at mucosal surfaces and should prove useful as biochemical tools for the study of sulphated glycoconjugates. PMID- 9218862 TI - Effect of psychoneural factors on intestinal epithelial function. AB - Stress has been associated with abnormal gastrointestinal function, including diarrhea and abdominal pain, and stress-associated gastric ulceration has frequently been documented. Stress can also exacerbate ongoing pathophysiology and often precedes relapses in patients with inflammatory bowel disease or irritable bowel syndrome. The relatively new field of psychoneuroimmunology is involved with the elucidation of mechanisms that explain the link between the central nervous system and immune-mediated pathophysiology. Recent progress examining the interaction among the nervous system, the immune system and the epithelium of the intestine is discussed, and the evidence for central nervous system control of this interaction is examined. PMID- 9218864 TI - Nausea, vomiting and diarrhea: an unusual presentation of multiple sclerosis. AB - The case of a young woman who presented with nausea, vomiting and diarrhea is outlined; the etiology turned out to be a first attack of multiple sclerosis. Plausible mechanisms are discussed. PMID- 9218865 TI - Dietary fads and gut mysteries versus nutrition with a grain of common sense. AB - Although nutritional self-help literature is directed at the general public, which usually allows the authors to evade critical review by the medical and scientific community, both doctors and lay people need to read with discernment and educated scepticism when major health claims are made. Many published claims are based in misconceptions and questionable logic, and it is important to be aware of the inconsistencies and wrong conclusions commonly found in dietary fads. Patients' questions and dietary practices over the past few years have helped the present authors become familiar with certain food fads and nutrition 'self-help' books, and develop responses to popular gut topics such as food allergies, food combinations and commercial food supplements. The authors also discuss whether fads can deliver on their promises and what to tell patients. PMID- 9218866 TI - The rise and fall of gastroenterology. PMID- 9218867 TI - Regulation of Myc-dependent apoptosis by p53, c-Jun N-terminal kinases/stress activated protein kinases, and Mdm-2. AB - Deregulated overexpression of c-Myc (Myc) confers susceptibility to apoptosis in several cell types, but the molecular regulation of these processes has not been well established. Here we have characterized several molecular changes that may modulate Myc-dependent apoptosis. Ectopic overexpression of Myc in both Rat1 fibroblasts and human osteosarcoma cells causes a dramatic increase of cellular p53 mRNA and protein, and this induction of p53 correlates with apoptosis triggered by withdrawal of serum. Stable transfection of a wild-type human p53 gene into Myc-transformed cells further potentiates apoptosis. Anticancer agents vinblastine and nocodazole also induce apoptosis in Myc-transformed Rat1 fibroblasts but are cytostatic to the same cells without Myc overexpression. We demonstrate that induction of Myc-dependent apoptosis in these cells is specifically associated with an activation of p46 c-Jun N-terminal kinase/stress activated protein kinase (JNK/SAPK) activity, whereas this JNK/SAPK activation is absent in stress-treated cells without Myc overexpression. Moreover, overexpression of the Mdm-2 gene in Rat1-myc cells significantly inhibits apoptosis induced by low serum but has little effect on apoptosis triggered by chemotherapeutic drugs. Interestingly, differential inhibition by Mdm-2 paralleled differential activation of p46 JNK/SAPK. Thus, our data support a functional involvement of p53 in Myc-dependent apoptosis and implicate potential regulatory roles for JNK/SAPK and Mdm-2 pathways in the regulation of apoptosis in Myc-transformed tumor cells. PMID- 9218868 TI - Differential regulation of human keratinocyte growth and differentiation by a novel family of protease-activated receptors. AB - Thrombin receptor (ThrR) and protease-activated receptor-2 (PAR-2) are members of a unique G protein-coupled receptor family, which are characterized by the unveiling of a tethered peptide ligand upon proteolysis of their NH2 terminus. We have previously shown that cultured human basal keratinocytes express both receptors (R.J. Santulli et al., Proc. Natl. Acad. Sci. USA, 92: 9151-9155, 1995); however, their functional role in epidermal physiology has yet to be described. In the present study, we determined the effects of receptor activation on keratinocyte cell growth and differentiation using thrombin (selective for ThrR), SLIGRL (selective for PAR-2), and SFLLRN (stimulates ThrR and PAR-2), as agonists. ThrR stimulation enhanced cell growth in a dose-dependent manner in the absence of growth factors (epidermal growth factor and bovine pituitary extract). In contrast, under the same conditions, activation of PAR-2 led to the inhibition of cell growth. This inhibitory activity by PAR-2 activation was also observed in the presence of growth factors. Activation of both receptors diminished protein expression of the differentiation marker transglutaminase type 1 induced by either calcium or IFN-gamma. Calcium-induced involucrin expression was also decreased. These results indicate that PAR-2 and ThrR differentially modulate keratinocyte function and may provide an important regulatory function in the epidermis by altering the functional state of keratinocytes. PMID- 9218869 TI - Mutational analyses of differentiation-dependent human papillomavirus type 18 enhancer elements in epithelial raft cultures of neonatal foreskin keratinocytes. AB - Human papillomaviruses (HPVs) reproduce only in differentiated squamous epithelia. Viral transcription is rather restricted in basal strata but increases dramatically in the spinous cells. Inopportune viral oncoprotein expression in the basal reserve cells can lead to dysplasias and carcinomas. Until now all studies to identify transcription factor binding sites within the upstream regulatory region (URR) that controls the expression of the oncogene have been conducted in proliferating cell cultures. We report the establishment of a reproducible and convenient system to examine cis elements important for differentiation-dependent transcriptional regulation. The bacterial lacZ gene under the control of the HPV URR-E6 promoter was transduced into primary human keratinocytes from neonatal foreskin by using high titer recombinant retroviruses. Acutely infected PHKs were then grown into stratified and differentiated epithelium on collagen rafts. lacZ expression was almost entirely restricted to the spinous cells, indicating that promoter activity was differentiation dependent, as seen in vivo. Using this system, we initiated a mutational analysis of previously identified promoter and enhancer elements within the HPV-18 URR. Three categories of mutation were observed: those that caused severe, moderate, or very small reduction in lacZ expressions. The results show both similarities and differences to previously published and present studies in proliferating primary human keratinocytes in monolayer cultures or in immortalized or transformed cell lines. This system is applicable to study both host and viral promoters that require squamous differentiation for their activity. PMID- 9218870 TI - Differentiation is inhibited and a senescence pathway is activated when simian virus 40 tsA 58-transformed human retinoblasts are grown at the restrictive temperature. AB - Neonatal human retina cells transformed by the SV40 tumor antigens were shown to leave the cell cycle and differentiate following treatment with agents that raise intracellular levels of cyclic AMP. This was true for both precrisis and immortal cell lines. However, with time, some of the differentiated retinoblasts withdrew neurites and returned to the cell cycle. Attempts to inhibit this process by developing cell lines transformed using SV tsA 58 with a temperature-sensitive phenotype for growth did not enhance but inhibited retinoblast-differentiating capacity. Growth restriction at the nonpermissive temperature was found to activate a senescence pathway. We propose that at the nonpermissive temperature, stable SV40 T-ag-p53 complexes fragment releasing p53, which transactivates p21waf1/cip1/sdi1 with the subsequent accumulation of p21 culminating in growth inhibition and senescence. PMID- 9218871 TI - Transcriptional defects underlie loss of E-cadherin expression in breast cancer. AB - Decreased expression of E-cadherin (E-cad), a calcium-dependent cell adhesion molecule, has been seen in many different epithelial cancers. Although somatic mutations in the E-cad gene have been identified in a small subset of tumors, in the majority of cancers, the mechanisms underlying loss of E-cad expression are poorly understood. We have cloned the human E-cad promoter and defined its critical components in functional assays. In eight human breast cancer cell lines, there was a striking correlation between endogenous E-cad gene expression and E-cad promoter activity observed following the introduction of reporter gene constructs into the lines. These and other observations suggest that defects in trans-acting pathways regulation E-cad expression are the primary basis for the loss of its expression in most breast cancers. The results have significant implications for understanding the gene expression differences that underlie tumor heterogeneity and progression events in breast and other epithelial cancers. PMID- 9218872 TI - 7-Hydroxystaurosporine (UCN-01) causes redistribution of proliferating cell nuclear antigen and abrogates cisplatin-induced S-phase arrest in Chinese hamster ovary cells. AB - A variety of agents, such as caffeine, have been shown to abrogate the DNA damage dependent G2 checkpoint and enhance cytotoxicity. However, these agents are too toxic for clinical use. We have reported that the potent protein kinase inhibitor 7-hydroxystaurosporine (UCN-01) at nontoxic doses abrogates the G2 arrest caused by the DNA-damaging agent cisplatin. Here, using Chinese hamster ovary cells, we show that cisplatin causes predominantly an S-phase arrest; UCN-01 abrogates this S-phase arrest, causing progression of cells to G2 and, subsequently, apoptotic cell death. In searching for an explanation for this accelerate DNA synthesis, we discovered that UCN-01 caused translocation of proliferating cell nuclear antigen (PCNA) to the detergent-insoluble, DNA-bound fraction. PCNA acts as a sliding clamp for DNA polymerase delta. Sequestering of PCNA by p21waf1/cip1 is required for p53-dependent G1 arrest in damaged cells. However, the S-phase arrest occurs independently of p53 and p21waf1/cip1. Our results suggest that PCNA is also a component of this S-phase checkpoint, despite the fact that CHO cells are defective for p53, and no increase in p21waf1/cip1 was observed. The mechanism by which PCNA is sequestered in the absence of p21waf1/cip1 and the mechanism by which UCN-01 disrupts this sequestration remain to be elucidated. PMID- 9218873 TI - Transcriptional regulation of intercellular adhesion molecule 1 by phorbol ester in human neuroblastoma cell line SK-N-SH involves jun- and fos-containing activator protein 1 site binding complex(es). AB - In this study, the regulatory elements involved in ICAM-1 transcriptional response to phorbol ester (12-0-tetradecanoylphorbol-13-acetate; TPA) have been investigated in the human neuroblastoma cell line, SK-N-SH. TPA induced intercellular adhesion molecule 1 (ICAM-1) protein expression in SK-N-SH cells within 24 h of treatment as judged by indirect immunofluorescence. Basal ICAM-1 mRNA levels were barely detectable in untreated SK-N-SH cells but were induced by TPA to a maximal level with 4 h and were reduced thereafter. Analysis of the 5' promoter sequence of ICAM-1 revealed two regions that functioned equally in the TPA induction of ICAM-1 transcription. The first region (-145 to -227) contained a nuclear factor-kappa B (NF kappa B) element. The second region (-316 to -390) contained a putative TPA-responsive element (TRE; TGATTCA) and a TATA box. Deletion and point mutation of the latter region indicated that the TRE was indeed the functional element within this region and acted fully and independently of all other elements including the TATA box at position -352. This TRE bound TPA induced specific nuclear complexes in vitro containing junD, c-jun, c-fos, and fra2 but not cAMP-responsive element binding/activating transcription factor family proteins. ICAM-TRE binding activity was induced within 30 min following TPA treatment. This preceded the appearance of ICAM-NF kappa B site binding activity. Cotransfection of c-jun and c-fos expression vectors into SK-N SH cells induced transactivation from ICAM-1 promoter constructs containing the intact but not mutated TRE site. Primer extension analyses revealed that TPA had induced transcription exclusively at two sites -40 and -41 bp upstream of the translation start site. These data show that the ICAM-TRE and its cognate jun- and fos-containing transcription factors play a predominant role in the transcriptional response of ICAM-1 to the protein kinase C activator TPA in SK-N SH cells. PMID- 9218874 TI - Induction of phosphorylation on BRCA1 during the cell cycle and after DNA damage. AB - BRCA1, the familial breast cancer susceptibility gene product, is a 220-kDA phosphorylated protein. BRCA1 immunoprecipitated from MCF7 cells blocked in G1-S phase or progressing through S-phase of the cell cycle migrated more slowly through SDS polyacrylamide gels than BRCA1 from cells maintained in serum supplemented media, serum-free media for 24 h, or delayed in G2-M phase by treatment with colchicine. Restoration of BRCA1 to the faster-migrating form, which occurred on release of cells from the G1-S-phase block, was prevented by the phosphatase inhibitor okadaic acid. Phosphatase treatment of immunoprecipitated BRCA1 resulted in the conversion of the slower-migrating form to the faster-migrating form. Although these results suggested that BRCA1 was preferentially hyperphosphorylated near the G1-S-phase boundary of the cell cycle, exposure of cells to DNA-damaging agents including UV light or treatment with hydrogen peroxide (H2O2) also promoted BRCA1 hyperphosphorylation. These same stimuli also eliminated the punctate nuclear staining pattern normally observed for BRCA1 in control cells. These results indicate that BRCA1 undergoes cyclic hyperphosphorylation during the cell cycle; however, this modification, as well as changes in BRCA1 nuclear staining, also occurs in response to DNA damage. PMID- 9218875 TI - Cyclin-dependent kinase activation and S-phase induction of the cyclin B1 gene are linked through the CCAAT elements. AB - Control of cell proliferation is dependent on the regulated expression of the cyclin genes. Induction of cyclin B1 gene expression in S phase has been shown to require sequences within the first 90 bp of the proximal promoter region. In this study, we defined the cell cycle regulatory elements within this region and explored the mechanism by which the cyclin B1 gene is activated. A CDE-like element that is important in S-phase regulation of other genes was not required for correct cell cycle expression of cyclin B1. Instead, two CCAAT boxes were essential for S-phase induction of cyclin B1 gene in both NIH3T3 and HeLa cells. Induction of cyclin B1 by cyclin/cyclin-dependent kinase (cdk) complexes were examined by cotransfection of the reporter along with appropriate expression vectors. Complexes of cdk4 with cyclin D1 or cdk2 with cyclin E or A can activate the cyclin B1 promoter, and activation is uniquely dependent on the CCAAT elements in both normal and heterologous contexts. This transcription factor NF-Y binds to both CCAAT elements. These findings suggest that S phase-specific induction of the cyclin B1 promoter is dependent upon NF-Y binding to the CCAAT elements and is correlated with activation by cyclin-dependent kinases. PMID- 9218877 TI - From genetics to prophylaxis. PMID- 9218878 TI - How to define the best efficacy parameters for migraine. PMID- 9218876 TI - Involvement of caspase family proteases in transforming growth factor-beta induced apoptosis. AB - Transforming growth factor-beta (TGF-beta) is a potent inducer of programmed cell death in liver as well as some hepatoma cell lines. To explore the mechanism by which TGF-beta induces apoptosis, we investigated the role of caspase family proteases in the apoptotic death of a human hepatoma cell line, Hep3B. We showed that TGF-beta-induced apoptosis was blocked by expression of the cowpox virus protein CrmA, a serpin-like pseudosubstrate for some of the caspase family proteases. CrmA expression, however, did not affect TGF-beta-induced regulation of promoter activities of the cyclin A and plasminogen activator inhibitor type I genes. These results indicate that CrmA inhibits a step specific for the apoptotic effect of TGF-beta. In addition to CrmA, a tripeptide caspase-protease inhibitor, z-Val-Ala-Asp-fluoromethylketone could also suppress TGF-beta-induced apoptosis in a dose-dependent manner. In TGF-beta-treated Hep3B cells, we observed a specific degradation of the catalytic subunit of DNA-dependent protein kinase, which was previously shown to be a substrate of caspase-3 but not several other members of the caspase family. This degradation was not seen in Hep3B cells transfected with CrmA nor in Hep3B cells pretreated with the tripeptide caspase inhibitor. Our study indicates a requirement of caspase family proteases in TGF beta-induced apoptosis. PMID- 9218879 TI - Is a central action of acute antimigraine drugs essential? PMID- 9218880 TI - Measuring central action of acute antimigraine drugs in humans. PMID- 9218881 TI - Would any acute treatment for migraine demonstrate recurrence? PMID- 9218882 TI - What is lacking in the treatment of paediatric and adolescent migraine? PMID- 9218883 TI - What is the role of international societies and what could they do better? PMID- 9218884 TI - Melanoma: stop the epidemic. PMID- 9218885 TI - A dermatologic diary. Portrait of a practice. PMID- 9218886 TI - Sargassum dermatitis. PMID- 9218887 TI - The sulfites: Part I. PMID- 9218888 TI - Hydrofluoric acid burns. AB - We observed seven patients who presented consecutively with hydrofluoric acid hand burns. The clinical characteristics of these patients and the course of their disease and treatment are reviewed. Dermatologists should be aware that a hydrofluoric acid burn constitutes a dermatologic emergency. Specialized treatment is required to prevent topical, system, and even lethal toxic effects. PMID- 9218890 TI - Generalized tuberous xanthoma with type IV hyperlipoproteinemia. AB - We report a case of generalized tuberous xanthoma with type IV hyperlipoproteinemia in a 42-year-old woman who had multiple, grouped, yellowish brown nodules bilaterally on the trunk, knees, elbows, palms, soles, and dorsal sides of multiple interphalangeal joints. On laboratory examination, serum triglyceride levels were elevated, and the pre-beta-lipoprotein band was increased on lipoprotein electrophoresis. Histopathologic findings showed many Touton giant cells and numerous foam cells in the dermis. PMID- 9218889 TI - A new triad: sensitivity to aspirin, allergic rhinitis, and severe allergic reaction to ingested aeroallergens. AB - Increasing attention has recently been paid to a group of patients who experience anaphylaxis after ingestion of foods prepared with mite-contaminated wheat flour. We present three cases of this syndrome, which occurs more often in young adults with allergic rhinitis and/or asthma. We have observed an increased frequency of sensitivity to aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs), manifested as urticaria or angioedema, in patients with this condition. PMID- 9218891 TI - Mees' lines in a patient with multiple parasitic infections. AB - Mees' lines, or transverse striate leukonychia, are classically associated with arsenic poisoning, but have been described in other cases of acute or chronic illness. Their pathogenesis is thought to be a disruption of nail plate keratinization secondary to systemic stress. Mees' lines are observed in a patient with helminthic and amebic infections and no history of arsenic exposure. This case demonstrates another clinical setting in which Mees' lines can appear, providing further evidence that Mees' lines may chronicle systemic disease. PMID- 9218892 TI - Aggressive giant keratoacanthoma of the face treated with intramuscular methotrexate and triamcinolone acetonide. AB - Keratoacanthoma is a common skin tumor characterized by rapid growth of a smooth dome-shaped nodule with a central plug of keratin, usually followed by spontaneous involution. We report the case of a giant keratoacanthoma on the face that continued to spread peripherally for several months despite vigorous therapeutic intervention. Treatment with methotrexate (25 mg intramuscularly) and Kenalog (triamcinolone acetonide, 40 mg intramuscularly) weekly finally slowed and stopped the expanding, deforming tumor growth and induced involution. PMID- 9218893 TI - Necrobiotic xanthogranuloma with paraproteinemia: an evolving presentation. AB - Necrobiotic xanthogranuloma with paraproteinemia is a progressive and destructive process that is often confused both clinically and histologically with other granulomatous and xanthomatous entities. It was first described by Kossard and Winkelmann in 1980. Prior to this, the entity was reported under a variety of names such as atypical multicentric reticulohistiocytosis with paraproteinemia, atypical xanthoma disseminatum, and atypical necrobiosis lipoidica. A 69-year-old woman experienced slightly pruritic and painful papules, plaques, and nodules. Initial biopsy specimens showed a granulomatous process consistent with granuloma annulare. Later biopsy specimens demonstrated histologic changes indicative of necrobiosis lipoidica. The most recent histologic findings are those of necrobiotic xanthogranuloma, with results of laboratory studies revealing a coexistent IgG kappa paraproteinemia. Patients with necrobiotic xanthogranuloma who demonstrate a benign monoclonal proteinemia and are evaluated for several years show a 9 to 11 percent risk of myeloma, amyloidosis, or macroglobulinemia. Multiple treatment regimens have been attempted, none of which are curative. We propose that this entity may be part of an evolutionary process that may start as a granulomatous entity and culminate in a xanthogranulomatous process with an accompanying paraproteinemia. PMID- 9218894 TI - Atrophodermia vermiculata. AB - We present a patient with atrophodermia vermiculata. A family tree study revealed an autosomal mode of inheritance with good penetrance. A slight improvement of the atrophic scars of the disease was noticed after local treatment with tretinoin cream, 0.05 percent, and cryotherapy. PMID- 9218895 TI - Cytologic features of neoplastic lesions in endocervical glands. AB - Cytologic criteria for classifying atypical endocervical cells on Pap smears are poorly defined. In this study we evaluated cytologic parameters that are useful in predicting the presence of neoplastic lesions (NL) and those that help distinguish squamous intraepithelial lesion (SIL) from glandular neoplastic lesions. The recently proposed Bethesda System (TBS) terminology for reporting atypical glandular cells of undetermined significance (AGUS) was also evaluated for its significance on patient management. Sixteen cases of biopsy-proven endocervical glandular NL that had cytologic smears available for review were included. Thirty-five smears with atypical endocervical cells and follow-up biopsies showing benign/reactive change (n = 22) and SIL involving glands (n = 13) were reviewed for comparison. Our results show that squamous NL often coexist with glandular NL. The presence of rosettes, hyperchromasia and increased N/C ratio is useful in distinguishing NL from benign/reactive conditions. Architectural features are helpful in distinguishing SIL from glandular NL. While a haphazard arrangement is more often seen with SIL, glandular NL are more likely to maintain polarity and to show glandular rosettes. Using TBS criteria, a conservative management seems justified in patients with AGUS-favor reactive and AGUS diagnosis on Pap smear, and colposcopy is indicated for patients with AGUS favor NL. PMID- 9218896 TI - Parotid fine-needle aspiration: a cytologic study of pediatric lesions. AB - Fine-needle aspiration (FNA) of parotid lesions in the pediatric age group presents interesting diagnostic problems. There is limited cytologic literature available on the subject. We studied the cytologic findings on FNA of parotid masses in seven such cases. Patients ranged in age from 1 to 17 years (mean = 10.4). Diagnoses included lymphoepithelial cyst (n = 1), mucoepidermoid carcinoma (n = 2), pilomatrixoma (n = 1), acinic-cell carcinoma (n = 1), rhabdomyosarcoma (n = 1), and Ewing's sarcoma (n = 1). Six cases were confirmed by subsequent histologic examination. The study elaborates on the cytologic features of each lesion and discusses the differential diagnosis of closely related entities peculiar to this age group. It highlights the role of FNA as the diagnostic modality of choice in dealing with pediatric parotid masses. PMID- 9218897 TI - Degree of dysplasia following diagnosis of atypical squamous cells of undetermined significance is influenced by patient history and type of follow-up. AB - Previous studies have shown that atypical squamous cells of undetermined significance (ASCUS) are a predictor of higher grade lesions when patients are followed up by biopsy. The purpose of this study was to examine follow-up on all patients with ASCUS to determine if the prediction of higher grade lesions seen in association with ASCUS is a function of bias in selection of patients for biopsy. The diagnosis of ASCUS, based on Bethesda System criteria, was made on 235 cases between June 1993 and December 1994 (2% of the total cases at Yale-New Haven clinics). Of these cases, 36 were biopsied and 94 were followed by cervical/vaginal smears (CVS). As has been seen in other retrospective studies, we found that 55% (20 of 36) biopsied after ASCUS on CVS showed condyloma or dysplasia. In cases with CVS follow-up instead of biopsy, only 26% (25 of 94) had significant lesions. We find that if ASCUS is evaluated by biopsy, the prediction for higher grade lesions is roughly twice that predicted by follow-up CVS. We purpose that nonstandardized management by gynecologists results, in many instances, in selection of patients with significant past history for biopsy. Consequently, the current literature finding of ASCUS as predictor of higher grade lesions should be reanalyzed, taking into account other parameters (past history, age, etc.) in the process of defining the clinical and biological implications of the ASCUS diagnosis. PMID- 9218898 TI - Reproducibility study of cervical cytopathology in Mexico: a need for regulation and professional accreditation. AB - Due to the high rate of false negative results in diagnosis of cervical cytopathology, in many countries its practice has been transformed through the application of several interventions aimed at medical regulation to improve diagnostic accuracy. Diagnostic reproducibility of gynecological cytopathology was evaluated in a series of 20 cytology specimens [Papanicolaou (Pap)] and 20 cervical biopsy. (CB) studies in different clinical stages, during 1994. The observation unit consisted of 30 pathologists who observed 2 groups of 20 Pap and 20 CB specimens. The standard was a cytopathologist certified by the Pathological Anatomy Council of Mexico. Intraclass reproducibility in gynecological cytopathology is low in Mexico. In a group analysis, concordance increased as clinical status of the cervical lesion increased. For moderate dysplasia, concordance in Pap was kappa = 0.04, compared to 0.23 in CB. Concordance of diagnosis of invasive cancer was 0.29 for Pap and 0.64 for CB. Using weighted kappa at the individual level for all possible diagnoses, concordance varied from 0.29 to 0.59 for Pap, and 0.42 to 0.65 for CB. The problem of reproducibility in cervical cytopathology in Mexico emphasizes the need for continuing education, uniform diagnostic criteria, and the advantages of a single operational classification-possibly the Bethesda System-since current classification systems are obsolete. PMID- 9218899 TI - Tall cell variant of papillary thyroid carcinoma: cytologic features and differential diagnostic considerations. AB - The cytologic features of the tall cell variant (TCV) of papillary thyroid carcinoma may be confused with those of other thyroid neoplasms with different prognoses and treatment modalities. Elucidation of the cytomorphology of this variant would be useful in planning treatment for this fairly aggressive variant of papillary carcinoma. The cytologic features of 20 cases of TCV were compared with those of 23 cases of the usual variant (UV) of papillary thyroid carcinoma and of 10 Hurthle-cell neoplasms (HCN). After a set of features was defined, the efficacy of employing it to distinguish TCV from UV and HCN was assessed by three cytopathologists (J.D.T., I.R., and M.O.), was independently examined 15 unknown cases selected by the first author. Aspirates of TCV showed some specific cytologic features which included large cell size with abundant granular cytoplasm and variably sized nuclei with granular chromatin. The cells were sometimes columnar, but more often were polygonal, and prominent cytoplasmic borders were present in 50% of cases. Intra-nuclear inclusions were more prominent in TCV than in UV. There was some overlap in the cytomorphology of some TCV and UV cases, and variable numbers of cells with UV features were encountered in TCV cases. Employing the cytologic features of TCV listed above, three cytopathologists examined the unknown cases, which included 7 cases of TCV, 4 cases of UV, and 4 cases of HCN. TCV was recognized as such by all three cytopathologists in 6 of 7 cases, and all UV and HCN were correctly typed by all three examiners. The cytologic features of TCV are sufficiently distinctive to enable separation from HCN and most cases of UV. Although the diagnosis of TCV may be rendered employing fine-needle aspiration biopsy, material, this diagnosis should be limited, in our opinion, to specimens which contain at least 30% of cells with typical TCV features. PMID- 9218901 TI - Fine-needle sampling of salivary gland lesions. III. Cytologic and histologic correlation of 75 cases of adenoid cystic carcinoma: review and experience at the Institut Curie with emphasis on cytologic pitfalls. AB - Fine-needle sampling (FNS) of 75 adenoid cystic carcinomas, including 44 primary tumors, 18 local recurrences, 10 lymph node, and 3 distant metastases, was performed in 66 patients. Concordant cytologic diagnoses were established in 68 tumors (90.7%), whereas 4 (5.4%) were classified as malignant (adenocarcinoma), 1 (1.3%) as suspicious, and 1 (1.3%) as pleomorphic adenoma. The material was insufficient for cytologic evaluation in 1 (1.3%) tumor. Adenoid cystic carcinoma is, in our opinion, easy to diagnose using the FNS technique. PMID- 9218900 TI - Fine-needle aspiration biopsy of gouty tophi: lessons in cost-effective patient management. AB - Gout, a disease resulting from the effects of hyperuricemia and a crystal-induced arthropathy, may produce soft tissue masses (tophi) which mimick neoplasia clinically and radiographically. We have recently diagnosed three cases of gouty tophus, two of which were clinically suspected to represent sarcomas, by fine needle aspiration biopsy (FNAB) after extensive radiologic and clinical evaluation. There were two women and one man. aged 71, 73, and 50 yr, with palpable soft tissue masses that involved the right forearm, right hand, and right foot, respectively, Biopsies were obtained by using 25-gauge needles without the aid of general anesthesia. Morphologically, aggregates and disassociated slender, needle-shaped crystals were abundant and easily recognized on both Diff-Quik and Papanicolaou stains. By using a polarizing microscope with a first-order red compensator, the crystals showed negative birefringence, characteristic of sodium urate. Benign-appearing histiocytes, foreign-body-type giant cells, neutrophils, and amorphous debris were scattered among the diagnostic crystals. The diagnosis of gouty tophus can be easily established with FNAB in conjunction with compensated polarizing microscopy. Application of FNAB in the initial evaluation of appropriate soft-tissue masses provides a cost effective diagnostic method, preventing more costly and often unnecessary clinical and radiologic tests. PMID- 9218902 TI - Adrenal histoplasmosis: diagnosis by fine-needle aspiration biopsy. AB - A 54-year-old man presented with a 6-month history of fever, night sweats, and weight loss. He had hepatosplenomegaly, and bilateral adrenal masses were discovered on computed tomographic (CT) scan. CT-guided fine-needle aspiration biopsy (FNAB) of the right adrenal mass demonstrated purulent material. Special stains done on this material showed organisms with morphologic features of Histoplasma capsulatum. The patient was started on antifungal therapy and discharged. FNAB of the adrenal gland is an effective method in the diagnosis of unusual infectious diseases. Special stains for micro-organisms proved helpful in the initial diagnosis of histoplasmosis. PMID- 9218903 TI - Breast and cutaneous mycobacteriosis: diagnosis by fine-needle aspiration biopsy. AB - The breast and skin are considered to be rare sites of extrapulmonary mycobacterial infection, comprising 0.1% to 0.5% of all tuberculosis cases, respectively. Fine-needle aspiration biopsy (FNAB) is a rapid and minimally invasive approach to diagnose extrapulmonary tuberculosis, and has been used successfully in identifying tuberculous lesions in the lymph nodes, thyroid, kidney, pancreas, vertebrae, and testis. Two cases of extrapulmonary mycobacteriosis diagnosed by FNAB are described: a 59-year-old Hispanic male with cutaneous mycobacterial infection of the head and neck region, and a 58-year-old white male with a unilateral tuberculous mastitis. In both instances, the FNAB material demonstrated acute neutrophilic exudate, few isolated aggregates of epithelioid histiocytes and lymphocytes, and on Fite-Farraco stain mycobacteria. Reported cases of tuberculosis diagnosed by FNAB have been few; this is the first case of cutaneous tuberculosis diagnosed by FNAB. PMID- 9218904 TI - Cytopathologic features of hepatic epithelioid hemangioendothelioma. AB - We present the cytological features of hepatic epithelioid hemangioendothelioma (EH), which is considered to be a vascular proliferation of intermediate malignant potential. The case report concerns a 52-yr-old previously healthy man discovered to have multiple hepatic masses upon evaluation for abnormal liver function tests. Fine-needle aspiration demonstrated a neoplasm composed of interanastomosing epithelioid cells that contained intracytoplasmic lumens. Histologic sections, immunohistochemistry, and ultrastructural evaluation were confirmatory. Although hepatic EH is a rare tumor, its characteristic cytological, histological, and ultrastructural features permit a straightforward diagnosis. It is important to distinguish this entity from adenocarcinoma and angiosarcoma because long-term disease-free survival is possible, especially in the setting of orthotopic liver transplantation. PMID- 9218906 TI - Fine-needle aspiration cytologic findings in a case of lymph node tumor of plasmacytoid monocytes. AB - We present a unique case of fine-needle aspiration (FNA) from a lymph node with subsequent histologic diagnosis of tumor of plasmacytoid monocytes (PMs). The patient had an associated myeloproliferative disease which terminated into an acute myelomonocytic leukemia. In a 95% ethanol-fixed, hematoxylin-eosin (H&E) stained smear, the tumor cells appeared monomorphic, medium size, with oval to indented nuclei, finely stippled chromatin, and small nucleoli; the cytoplasm was scanty and slightly eccentric. The cytologic picture suggested some subtypes of small-cell non-Hodgkin's lymphomas or a leukemic infiltration. In the FNA specimen, the cells appeared immunocytochemically negative for some B- and T-cell markers (MB2, L26, LN1, and UCHL1) and strongly positive to KP1, a known histyocyte-macrophage and myeloid marker. The FNA differentiated diagnoses are discussed. PMID- 9218905 TI - Cytologic diagnosis of fat emboli in peripheral blood during sickle cell infarctive crisis. AB - A diagnosis of fat emboli can be suspected in a patient presenting with the typical symptoms of the fat embolism syndrome, but is rarely proved pathologically, except at autopsy. We described a 25-yr-old man with sickle cell anemia who developed an infarctive crisis complicated by unexplained fever, neurologic change, and respiratory abnormalities. Blood drawn from the femoral vein and examined cytopathologically yielded necrotic bone marrow elements admixed with fat. The cytologic finding of fat emboli from necrotic bone marrow provided the diagnosis and helped guide subsequent medical intervention. This sample test is recommended for patients at risk for fat emboli to aid in the clinical diagnosis. PMID- 9218907 TI - Thymoma mimicking thyroid papillary carcinoma: another pitfall in fine-needle aspiration. AB - Thymomas are composed of a mixture of epithelial and lymphoid cells. When the lymphoid cells predominate, the possibility of malignant lymphoma must be considered. Recently, a thymoma mimicking lymphoblastic lymphoma was reported in this journal as a diagnostic pitfall in a mediastinal aspiration. We encountered another diagnostic pitfall at the other end of the spectrum: a thymoma with a dominant epithelial component in which a papillary carcinoma of thyroid had to be considered in the differential diagnosis. PMID- 9218908 TI - Classics in cytology. VIII: Diffuse meningeal sarcomatosis with invasion of the spinal cord and the spinal roots. Positive cytological results, especially of the cerebrospinal fluid. 1904. PMID- 9218909 TI - False-negative diagnosis in fine-needle aspirations of squamous-cell carcinoma of head and neck. AB - Squamous-cell carcinoma (SCC) is the most common malignancy encountered in the head and neck area. Fine-needle aspiration (FNA) is routinely performed in the diagnosis of primary, recurrent, and metastatic SCC. This paper describes several diagnostic problems and pitfalls that might be seen in FNAs of SCC. These include cystic changes, well-differentiated SCC, spindles SCC, and SCC with foreign-body giant cells, keratin plaques, and ghost cells. Recognition of these patterns along with clinical correlation enables the pathologist to prevent a false negative diagnosis. PMID- 9218910 TI - Immunocytochemistry controls using cell culture. AB - A continuing problem in cytology laboratories is the lack of adequate control material for immunocytochemical testing. Usual control procedures involve testing paraffin-embedded control materials along with the patient specimens. These control materials are fundamentally unlike cytologic preparations. We have developed a method to make control preparations for immunocytochemical analysis using cultured anaplastic cells with known antigenic features from commercial sources. Cell lines included melanoma, rhabdomyosarcoma, T-cell leukemia, and squamous-cell carcinoma. Modified Saccomano and acetone fixation coupled with the cytospin technique enabled good-quality preparations. Cell lines were tested with antibodies for HMB-45, actin, leukocyte common antigen (LCA) and cytokeratin, which avidin-biotin immunoperoxidase and diaminobenzidine (DAB) as chromogens. Our final preparations were easily interpretable with excellent morphologic preservation of cellular detail. Cultured cells provide a superior method for preparing almost unlimited numbers of control slides for immunocytochemistry for laboratories with access to a tissue culture facility. PMID- 9218911 TI - Intraoperative touch-imprint cytological diagnosis of follicular variant of papillary thyroid carcinoma. PMID- 9218912 TI - Stability of Legionella urinary antigens over time. AB - Twenty-two urine samples positive for Legionella pneumophila serogroup 1 antigen by EQUATE radioimmunoassay (RIA) (Binax, Portland, ME, USA) were stored at various temperatures and the RIA repeated at 1, 7, 30, 90, and 120 days to evaluate stability of the urinary antigens. The mean ratios of patient/negative control remained stable. Although there was a 10% decrease in the mean ratios after 1 month, changes were not significant. However, individual samples with ratios close to 3 may fall to < 3. PMID- 9218913 TI - Microbiologic and pharmacodynamic principals applied to the antimicrobial susceptibility testing of ampicillin/sulbactam: analysis of the correlations between in vitro test results and clinical response. AB - The correlation between various ampicillin/sulbactam in vitro antimicrobial susceptibility test results and the clinical outcome of patients treated with this agent have been examined. A survey of over 29,000 clinical isolates of the family Enterobacteriaceae found that the proportion of susceptible pathogens as assessed by current susceptibility testing interpretive guidelines (NCCLS) for disk diffusion and dilution (MIC) assays was significantly less than the proportion of patients cured or clinically improved in ampicillin/sulbactam clinical trials. Also, the results of two NCCLS methods differ greatly in the perceived percentages of susceptible strains (63.9% versus 72.2%; unacceptable variation). Furthermore, the current interpretive criteria resulted in high false susceptible (4.2%) and total (19.7%) error rates. When proposed interpretive guidelines were applied, approximately 73 to 87% of the Enterobacteriaceae strains were observed to be susceptible, the variation between methods was minimized, and the error rates were reduced. A retrospective analysis of data from clinical trials with ampicillin/sulbactam indicated that the proportion of patients who were cured or clinically improved and bacterially eradicated was not appreciably different in patients having baseline Enterobacteriaceae pathogens with MICs of 16 or 32 micrograms/ml (ampicillin MIC component) as compared to those with pathogens having MICs of < or = 8 micrograms/ml. Studies in animals, in vitro models, and pharmacokinetic considerations indicate that a change in the MIC breakpoint for ampicillin/sulbactam should be considered. The proposed interpretive guideline revisions for ampicillin/sulbactam susceptibility testing of the Enterobacteriaceae were 1) use current diagnostic reagents with criteria of < or = 16/8 micrograms/ml (> or = 14 mm) as susceptible and > or = 64/32 micrograms/ml (< or = 10 mm) as resistant; e.g., 75.9 to 76.0% spectrum and 1.3% false-susceptible error; 2) use alternative diagnostic reagents (1:1 ratio MIC; 20/20 micrograms disks) with criteria of < or = 8/8 micrograms/ml (> or = 18 mm) as susceptible and > or = 32/32 micrograms/ml (< or = 14 mm) as resistant; e.g., 73.3 to 76.9% spectrum and 1.8% false-susceptible error; or 3) use alternative diagnostic reagents with criteria of < or = 16/16 micrograms/ml (> or = 14 mm) as susceptible and > or = 64/64 micrograms/ml (< or = 10 mm) as resistant; e.g., 84.7 to 86.9% spectrum and 1.3% false-susceptible error. Data from a comprehensive in vitro survey of clinical isolates, retrospective analyses of clinical trials, and studies of animal models support the modification of contemporary interpretive guidelines for ampicillin/sulbactam antimicrobial susceptibility tests. The best short-term criteria would apply current in vitro diagnostic reagents and a modified susceptible breakpoint (< or = 16/8 micrograms/ml as susceptible; option 1 above) until new diagnostic reagents can be qualified by means of studies needed for quality assurance of standardized methods (NCCLS M23-A and FDA procedures). These changes would provide a better in vitro prediction of ampicillin/sulbactam efficacy in clinical practice. PMID- 9218914 TI - A simple polymerase chain reaction technique to detect and differentiate Shigella and enteroinvasive Escherichia coli in human feces. AB - A simple polymerase chain reaction (PCR) procedure using IS630-specific primers was developed as a general diagnostic probe to detect Shigella and enteroinvasive Escherichia coli (EIEC). However, IS630 and the other two previously reported molecular probes, ipaH and ial, cannot be used to differentiate among Shigella serotypes and EIEC strains that cause dysentery. The sensitivity of PCR protocol was determined to be 100-200 shigellae for each PCR reaction. An enrichment incubation would allow the detection of shigellae in stool samples with low bacterial concentration; i.e., < 10(4) CFU/gram. Serotype-specific primers derived from the rfc genes of differentiate among Shigella serotypes in the laboratory, such as S. sonnei, S. flexneri, and S. dysenteriae 1. It was demonstrated further that the multiplex PCR system containing rfc-specific primers can efficiently identify the most prominent Shigella serotypes in raw stool samples of acute diarrheal patients. PMID- 9218915 TI - Reproducibility of broth microdilution and disk diffusion susceptibility tests of nine antimicrobial agents against Streptococcus pneumoniae ATCC 49619. AB - Collaborative studies documented the reproducibility of broth microdilution susceptibility tests of Streptococcus pneumoniae against nine antimicrobial agents and of disk diffusion tests with six of those drugs. Replicate tests of Streptococcus pneumoniae ATCC 49619 in five different laboratories led to the following provisional quality control limits: cefdinir--0.03 to 0.25 microgram/ml and 26 to 31 mm; cefetamet--0.5 to 2 micrograms/ml and 20 to 25 mm; ciprofloxacin -0.25 to 1 microgram/ml and 20 to 26 mm; clinafloxacin--0.03 to 0.125 microgram/ml and 28 to 34 mm; grepafloxacin--0.06 to 0.5 microgram/ml and 21 to 28 mm; PD131628--0.125 to 0.5 microgram/ml and 24 to 29 mm; clindamycin--0.03 to 0.12 microgram/ml; cefpodoxime--0.03 to 0.12 microgram/ml; and trospectomycin--1 to 4 microgram/ml (disk tests were not evaluate for the latter three drugs). PMID- 9218916 TI - Comparison of the Vidas and Bio-Whittaker enzyme immunoassays for detecting IgG reactive with varicella-zoster virus and mumps virus. AB - A total of 215 sera, 164 positive and 51 negative for antibody reactive with varicella-zoster virus (VZV), were analyzed using two commercially available enzyme immunoassays (EIAs) for detecting VZV-IgG, the Vidas Varicella-Zoster IgG assay (bioMerieux Vitek, Inc., Hazelwood, MO, USA) and the Bio-Whittaker Varicella II assay (Bio-Whittaker, Inc., Walkersville, MD). The sensitivity and specificity of the Vidas and EIA for VZV-IgG was 98.9 and 100%, respectively. The sensitivity of the Bio-Whittaker VZV-IgG EIA assay was 98.8%, specificity 96.1%. Equivocal results were obtained with 4 and 2 sera, respectively. A total of 185 sera, 169 positive and 16 negative for mumps virus antibody, were analyzed with Vidas and Bio-Whittaker mumps IgG EIAs. The sensitivity of the two assays for detecting mumps IgG was 99.4% and 95.7%, respectively. Both assays demonstrated 100% specificity. Two of the 185 sera tested for mumps antibody yielded equivocal results with the Vidas EIA; four equivocal results were obtained with the Bio Whittaker EIA. PMID- 9218917 TI - Critical evaluation of the Vitek GNS F6 card results compared to standardized, reference susceptibility test methods. AB - A large number of Enterobacteriaceae (291 unselected and 30 clinical challenge isolates) were used to evaluate the accuracy of the Vitek GNS-F6 susceptibility testing card for 10 antimicrobial agents (ampicillin, ampicillin/sulbactam, aztreonam, ciprofloxacin, imipenem, mezlocillin, ofloxacin, piperacillin, ticarcillin, and ticarcillin/clavulanate). Results were compared to reference broth microdilution and disk diffusion methods. Highest interpretive error on initial processing were observed with ticarcillin/clavulanate (very major false susceptible error, 3.4%), imipenem (major false-resistant error, 1.9%), and ampicillin/sulbactam (minor error, 15.9%). Repeat testing resolved many very major errors (21 of 41 results), and the overall accuracy rate was improved from 92.1 to 93.4%. Analysis of all minor interpretive errors demonstrated that the trend for Vitek was to report slightly lower MICs (6 of 10 drugs; 97 of 159 results) in comparison to the reference test results. If a heavy inoculum was used in the Vitek cards, false resistance was observed more frequently with aztreonam and penicillins. These data demonstrate that susceptibility testing accuracy with Vitek GNS-F6 card based on categorical agreement varies from 83.2% (ampicillin/sulbactam) to 99.4% (ciprofloxacin) when testing enteric bacilli. Some antimicrobial agents (beta-lactamase inhibitor/penicillin combinations, antipseudomonal penicillins) may require slight modification of interpretive software. Finally, the overall accuracy of the Vitek System was greater than 91% for 9 of 10 drugs tested with a very low, acceptable rate of false-susceptible error (0.8%). PMID- 9218918 TI - Duration of fungal culture incubation in an area endemic for Histoplasma capsulatum. AB - To determine the optimum incubation period for recovery of fungi from clinical specimens in an area where Histoplasma capsulatum is endemic, we reviewed the results of 4259 consecutive fungal cultures. Of the total 1306 fungi isolated, 92.6% were detected by day 7, and 97.9% were detected by day 14. For the 1027 yeast isolates, only 1.4% were detected after day 7, none of which was considered clinically significant. Of the 281 mould isolates, 91.1% were detected by day 14; and of those detected after day 14, only isolates of H. capsulatum were considered clinically significant. An incubation protocol based on type of specimen is suggested. PMID- 9218919 TI - Comparison of BACTEC 12B vs solid media for the recovery of Mycobacterium avium complex from blood cultures in AIDS patients. AB - We compared liquid (BACTEC 12B) and solid culture media for the diagnosis of Mycobacterium avian complex (MAC) bacteremia among 258 AIDS patients with a positive blood culture. Neither culture media alone had adequate sensitivity; BACTEC 12B detected growth earlier. Use of both liquid and solid media may improve the yield of mycobacterial blood culture. PMID- 9218920 TI - Comparison of Etest and agar dilution for testing the activity of three macrolides against Bordetella parapertussis. AB - Agar dilution and Etest using Mueller-Hinton II agar supplemented with 5% whole defibrinated horse blood were compared for testing the activities of azithromycin, clarithromycin, and erythromycin against 34 clinical isolates of the fastidious species Bordetella parapertussis. There was good overall agreement (100.0% within +/- 1 log2 dilution step) between both methods. The Etest offers an excellent and convenient alternative to agar dilution for testing the activities of macrolides against B. parapertussis. PMID- 9218921 TI - "Academic drug-detailing": from project to practice in a Swedish urban area. AB - OBJECTIVES: To develop and test the long-term feasibility of an interdisciplinary independent drug information service providing both written and oral drug information to physicians in an urban area of Sweden (> 400,000 inhabitants). METHODS: A drug information service was developed encouraging a cooperative approach between a department of clinical pharmacology, general practitioners (GPs), pharmacists, and Drug and Therapeutic Committees. Scientifically-based drug information was condensed and interpreted by a team and presented in both written and oral form. In one part of the area, both oral and written information was provided, while in another part of the area, only written information was distributed. Questionnaires and one prescription survey were performed to elucidate the knowledge and attitudes of the GPs regarding drug treatment of one condition (urinary tract infection, UTI, and norfloxacin were used as examples), as well as their opinion of our services. RESULTS: Over a period of 10 years, 75 issues of a drug bulletin (2000 copies) were distributed. Oral producer independent drug information, provided jointly by a GP and a pharmacist, was given on 16 occasions in each of 30 health centres (150 GPs). Around 80% of the GPs participated in the meetings. Of these GPs, 75% found the service important for their daily work. A majority of the GPs had prescribed the test drug, norfloxacin, not a first-line drug according to local recommendations, 1 year after approval. A significantly lower proportion of prescribers were observed in the area where the GPs had been provided with both written and oral information regarding recommended treatment (including first-line drugs) for uncomplicated cystitis. The approximate cost for this service in 1995 was SEK 0.685 million (USD 0.1 million); the prescribing costs of the 150 GPs were estimated at SEK 255 million per year. This means that the cost of the service per GP is only around 0.3% of normal prescribing costs. CONCLUSION: Over a period of 10 years the information/education method described here has proven sustainable and feasible in terms of providing the information, regarding participation of the target group GPs in the oral sessions, and regarding integration of the service into the existing health care system. PMID- 9218922 TI - Valsartan, a new angiotensin II antagonist for the treatment of essential hypertension: efficacy and safety compared to hydrochlorothiazide. AB - OBJECTIVE: To compare the antihypertensive efficacy of a new angiotensin II antagonist, valsartan, with a reference therapy, hydrochlorothiazide (HCTZ). METHODS: In this double-blind study, 167 adult out-patients with mild-to-moderate essential hypertension were randomly allocated in equal number to receive valsartan 80 mg or HCTZ 25 mg for 12 weeks. In patients whose blood pressure (BP) remained uncontrolled after 8 weeks of monotherapy, atenolol 50 mg was added to the initial treatment. Patients were assessed at 4, 8 and 12 weeks. The primary efficacy variable was change from baseline in mean sitting diastolic BP (SDBP) at 8 weeks. Secondary variables included change in sitting systolic BP (SSBP) and responder rates (percentage of patients with SDBP < 90 mmHg or drop > or = 10 mmHg compared to baseline) at 8 weeks. RESULTS: Valsartan and HCTZ were both effective at lowering diastolic and systolic blood pressure at all time points. Similar falls were seen in both groups with no significant differences between treatments. For the primary variable (decrease in SDBP) there was no significant difference between treatments. For SSBP there was also no significant difference observed. Responder rates at 8 weeks were 74% for valsartan and 62% for HCTZ (P = 0.10). Both treatments were well tolerated, both as monotherapy, and when combined with atenolol 50 mg per day. CONCLUSION: The data show valsartan 80 mg to be as effective as HCTZ in the treatment of mild-to-moderate hypertension. The results also show valsartan to be well tolerated when taken alone or in combination with atenolol. PMID- 9218923 TI - Fibrinolytic treatment in suspected acute myocardial infarction--use and risks in clinical practice. AB - OBJECTIVES: To study the adherence to guidelines concerning fibrinolytic treatment of patients with suspected myocardial infarction and to obtain information on severe events in clinical practice. METHODS: Prospective reporting of all patients admitted for suspected acute myocardial infarction during 4 months in 1994 from 69 (73.4% of all) Swedish coronary care units. RESULTS: The study covers 10,652 admissions, representing 9726 patients. The mean percentage treated with fibrinolytic drugs of patients with a positive ECG (ST-elevation and/or bundle branch block), a delay < 12 h and no contraindications was 56%. The interhospital range was 18.1-94.1%. Fibrinolytic drugs were given with a delay time > 24 h in 12.5% of women and 15.7% of men, and 36.1% of patients with verified acute myocardial infarction were given fibrinolytic drugs. Streptokinase was used in 82.7% and alteplase in 15.5% of the patients, respectively (interhospital range 0-53.3%). CONCLUSIONS: Fibrinolytic therapy seems to be used in a non-rational way at several hospitals. Local quality systems may be a way to assure better care. PMID- 9218924 TI - Drug-induced thrombocytopenia: clinical data on 309 cases and the effect of corticosteroid therapy. AB - OBJECTIVE: To analyse the clinical picture and the course of thrombocytopenia induced by non-cytotoxic drugs, and to evaluate a possible therapeutic effect of corticosteroids. METHODS: A retrospective analysis of 309 well-documented cases of drug-induced thrombocytopenia was performed. Data sources were reports from the files of the Danish Committee on Adverse Drug Reactions and discharge summaries. RESULTS: The median length of exposure to the offending drug, before development of thrombocytopenia, was 21 days. The median nadir platelet count was 11 x 10(9).l-1, and 74% of the patients had clinical haemorrhage. Bone marrow examination generally showed hyperplastic reactive changes and a variable number of megakaryocytes. Slight leucopenia was present in 6% of the patients and 16% were anaemic. Complete recovery was seen in 87% of cases, with a median recovery rate of 8 days. The standard treatment was corticosteroids, which were administered in 53% of the cases. No difference in recovery between corticosteroid-treated and untreated patients was observed. No other clinical parameter affected the recovery rate. The mortality rate due to haemorrhage was 3.6%. CONCLUSION: Thrombocytopenia induced by non-cytotoxic drugs is characterised by heterogeneous clinical picture and recovery is generally rapid. Although corticosteroids seem inefficient, we still recommend that severe symptomatic cases of drug-induced thrombocytopenia are treated as idiopathic thrombocytopenic purpura due to the difficult initial differentiation between the two conditions. PMID- 9218925 TI - Prostaglandin E1 decreases human arterial accumulation of radiolabeled apo B containing lipoproteins in vivo. AB - OBJECTIVE: An increased apo B-containing lipoprotein influx and cholesterol ester accumulation in arteries are well-known events in human atherogenesis. In vitro and experimental animal studies have provided evidence of a beneficial effect of PGE1 on both vascular apo B-containing lipoprotein accumulation and cholesterol ester content. METHODS: We examined the effect of PGE1 (administered via an intravenous portable infusion pump at a rate of 5 ng PGE1 kg-1.min-1 for 5 days a week, 6 h daily, over a total of 5 weeks) in ten patients (eight males, two females) on 123I-apo B-containing lipoprotein accumulation into the large arteries in vivo. Apo B-containing lipoprotein isolation was carried out by immunoaffinity chromatography and radiolabeling with the iodine monochloride method. 123I-apo B-containing lipoprotein accumulation was imaged and quantified by means of special computer software before and after 5 weeks of PGE1 therapy. RESULTS: PGE1 led to a significant decrease in maximal arterial apo B-containing lipoprotein retention. The mean decrease in the carotid and femoral arteries in type I lesions amounted to between 16.9% and 30.4%, and in type II lesions between 22.4% and 30.7%, 20 h after injection of radiolabeled apo B-containing lipoprotein. The type of arterial apo B-containing lipoprotein kinetic curves, however, remained unchanged. CONCLUSION: These findings indicate that PGE1 decreases the apo B-containing lipoprotein influx in the large arteries and the vascular cholesterol content, suggesting that PGE1 may lead to regression of lipid-rich lesions in human in vivo. PMID- 9218926 TI - Cytotoxic activity of cyclosporin A and [3-keto-Bmt1]-[Val2]-cyclosporin (SDZ PSC 833) on tumour cells from patients with haematological malignancies. AB - OBJECTIVE AND METHOD: The fluorometric microculture cytotoxic assay was employed for characterisation of the cytotoxic effect of cyclosporin A (CsA) and its non immunosuppressive analogue SDZ PSC 833, [3-keto-Bmt1]-[Val2]-cyclosporin (PSC) in tumour cells from patients with haematological or solid tumours. RESULTS: Tumour cells from patients with chronic lymphocytic leukemia (CLL) or non-Hodgkin's lymphoma (NHL) were found to be more sensitive to both drugs than those of tumour cells from patients with acute lymphocytic leukemia (ALL), acute myoblastic leukaemia (AML) and various solid tumours. There was a close correlation between the effects of the two drugs (correlation coefficient 0.71), but CsA was slightly more active than PSC in most diagnoses. No tumour cells sample showed sensitivity to PSC without also being sensitive to CsA. There was a moderate level of correlation between the activity pattern of CsA and doxorubicin (correlation coefficient 0.66), whereas the correlations with other cytostatics, such as vincristine, cytarabine and melphalan, were low (correlation coefficient -0.11 to 0.33). CONCLUSION: The results indicate that PSC shares the direct cytotoxic properties of CsA, but is slightly less potent. Clinical testing of the cytotoxic effect of these agents in haematological malignancies seems warranted and the apparent non-cross-resistance with standard agents makes cyclosporins a potentially useful adjunct to chemotherapy in those diagnoses. PMID- 9218927 TI - Changes in the pattern of antidepressant use upon the introduction of the new antidepressants: a prescription database study. AB - OBJECTIVE: To study whether the newer antidepressants have changed the patterns of antidepressant use, and whether the claimed better adverse effect profile of the lower antidepressants is reflected in their use as monitored by a prescription database. METHOD: By means of a prescription database (OPED), the use of antidepressants from 1991 to 1993 in Odense, Denmark, was analysed. RESULTS: The 1-year prevalence of antidepressant use increased significantly from 1.60% to 2.00%, which still is below the claimed 1-year prevalence of depression of at least 5%. The increase was mainly due to a rapidly increasing use of the newer antidepressants, accompanied by a moderate decline in the use of older antidepressants (mainly tricyclic antidepressants). The patterns of antidepressant use were very polymorphic, with about 5% being on continuous use for all 3 years and groups of each 20-30% being treated with: (1) several series or (2) one series or (3) only by one prescription. The share of patients presenting only one prescription (20%) was the same for older and newer antidepressants. Likewise, the rate of shifts from older to newer antidepressants or vice versa was the same (7% vs 6%). The duration of treatment did not differ much between older and newer antidepressants. Relative to the defined daily dose (DDD), the older antidepressants were given in much lower doses (median 0.63 DDD) than the newer antidepressants (median 1.05 DDD). CONCLUSION: It is concluded that many depressed patients are still not receiving antidepressant treatment and that the claimed better adverse effect profile of the newer antidepressants was not clearly reflected in their use. PMID- 9218928 TI - Population pharmacokinetics of caffeine in premature neonates. AB - OBJECTIVE: To determine population pharmacokinetic parameters of caffeine in premature neonates. METHODS: This population analysis was done using 145 serum concentration measurements gathered from 75 hospitalized patients during their routine clinical care. The data were analysed by use of NONMEM (mixed effects modeling) according to a one-compartment open model with either zero or first order absorption and first-order elimination. The effect of a variety of developmental, demographic and clinical factors (gender, birth weight, current weight, gestational age, postnatal age, postconceptional age and concurrent treatment with phenobarbital and parenteral nutrition) on clearance and volume of distribution was investigated. Forward selection and backward elimination regression identified significant covariates. RESULTS: The final pharmacostatistical model with influential covariates were as follows: clearance (m1.h-1) = 5.81-current weight (kg) + 1.22.postnatal age (weeks), multiplied by 0.757 if gestational age < or = 28 weeks and 0.836 if the current primary source of patients' nutrition is parenteral nutrition, and volume of distribution (ml) = 911.current weight (kg). The inter-individual variability in clearance and the residual variability, expressed as coefficients of variation, were 14.8%, and 18.44%, respectively. Due to the lack of information on the data set we were unable to characterize the interindividual variability for volume of distribution. CONCLUSION: In this study, which involved on average only two serum concentrations of caffeine per patient, the use of NONMEM gave us significant and consistent information about the pharmacokinetic profile of caffeine when compared with available bibliographic information. Additionally, parenteral nutrition and low gestational age (< or = 28 weeks) may even come to be considered as risk factors, and their presence may serve as an indicator of the need for periodic monitoring of caffeine concentrations in premature infants. PMID- 9218929 TI - Interaction between carbamazepine and bromperidol. AB - OBJECTIVE: The interaction between carbamazepine and bromperidol was studied in 13 schizophrenic inpatients. METHODS: Before carbamazepine addition, the subjects were taking bromperidol 12-24 mg.day-1 for 1-20 weeks. Carbamazepine 400 mg.day-1 was coadministered for 4 weeks, and blood samplings were performed before carbamazepine addition and at weekly intervals after the addition. Plasma concentrations of bromperidol and its reduced metabolite were measured by high performance liquid chromatography. RESULTS: Carbamazepine significantly decreased plasma concentrations of both bromperidol and reduced bromperidol for all weeks. On average, the plasma concentrations of bromperidol and reduced bromperidol at 4 weeks were 37% and 23% of the corresponding precarbamazepine values. Despite these decreases in plasma concentration, the Clinical Global Impression scores decreased slightly but significantly after carbamazepine addition. CONCLUSION: The present study suggests that carbamazepine decreases plasma concentrations of bromperidol and its reduced metabolite by inducing the metabolism of these compounds. Nevertheless, adjunctive carbamazepine may be useful for schizophrenic patients treated with bromperidol. PMID- 9218930 TI - Pharmacokinetics of sertindole and dehydrosertindole in volunteers with normal or impaired renal function. AB - OBJECTIVE: To study the effect of renal impairment on the pharmacokinetics of sertindole. METHODS: A single 4 mg oral dose of sertindole was given to normal subjects (n = 6) and subjects with various degrees of impaired renal function (n = 18) classified into mild, moderate, and severe/hemodialysis based on their creatinine clearance). The relationships between the pharmacokinetic parameters and the degree of renal impairment were investigated using regression analysis with creatinine clearance as an explanatory variable along with body weight. Subjects were also genotyped for CYP2D6-A or 2D6-B mutations. RESULTS: The mean CL/f and t1/2 values of sertindole ranged from 14 to 31 1.h-1 and from 73 to 93 h, respectively, and were not significantly related to creatinine clearances. There was no indication of any influence of creatinine clearance on the fraction of sertindole (0.994-0.995) binding to plasma proteins. The total fraction of the sertindole dose removed by dialysis was less than 0.1% Subjects with B/B genotype (n = 2) for CYP2D6 were associated with a distinctly lower clearance of sertindole (6.3 vs 25.3 1.h-1) than subjects with wt/wt genotype for CYP2D6. CONCLUSIONS: Since the pharmacokinetics of sertindole are unchanged by renal impairment, dosage adjustment does not appear to be necessary for subjects with various degrees of renal insufficiency or subjects with renal failure requiring hemodialysis. PMID- 9218931 TI - Pharmacokinetics and pharmacodynamics of ranitidine in renal impairment. AB - OBJECTIVE: The pharmacodynamics and pharmacokinetics of ranitidine were examined in subjects with varying degrees of renal function to determine the effect of this condition on acid-antisecretory activity. METHODS: Subjects with creatinine clearances (Ccr) ranging from 0 to 213 m1.min-1 received single 50-mg and 25-mg i.v. doses of ranitidine. This was followed by determination of serum and urine ranitidine concentrations, and continuous gastric pH monitoring for 24 h. RESULTS: Serum ranitidine concentrations were described by a two-compartment model linked to a sigmoidal Emax model describing gastric pH. Ranitidine renal clearance, ranging from 0 to 1003 m1.min-1, correlated with CPAH (r2 = 0.707), while non-renal clearance was unaltered. Steady-state volume of distribution decreased by half in severe renal impairment. No changes in the effective concentration at half-maximal response (EC50), maximal response (Emax), or basal response (E0) were observed. Thus, renal elimination of ranitidine declined in parallel with renal function, while sensitivity to the pharmacologic effect (gastric pH elevation) was unaltered. Ranitidine was cell tolerated in these renally impaired subjects. CONCLUSION: These data indicate that the current recommendation for renal impairment dose reduction (by two-thirds when Ccr < 50 m1-min-1) might result in under-treating moderately impaired patients, and suggests a less conservative dose reduction (by half when Ccr < 10 m1.min-1) to avoid therapeutic failure while remaining within the wide margin of safety for this drug. PMID- 9218932 TI - Influence of an acidic beverage (Coca-Cola) on the absorption of itraconazole. AB - OBJECTIVE: To evaluate the effectiveness of Coca-Cola in enhancing the absorption of itraconazole. METHODS: Eight healthy volunteers were randomized to receive two treatment sequences in a two-way crossover design with a 1-week wash-out period separating each study treatment. Treatment 1, the control, consisted of 100 mg itraconazole with 325 ml water. Treatment II was identical to treatment I, except that itraconazole was administered with 325 ml of Coca-Cola (pH 2.5). RESULTS: Serum itraconazole concentrations, after administration with Coca-Cola (treatment II), were higher than after administration with water (treatment I). The mean AUC was 1.12 vs 2.02 micrograms.h.m1-1, the mean Cmax was 0.14 vs 0.31 micrograms.m1 1 and the mean tmax was 2.56 vs 3.38 h in treatments I and II, respectively. CONCLUSION: The absorption of itraconazole can be enhanced by Coca-Cola. PMID- 9218933 TI - Comparative pharmacology between habitual and non-habitual coffee-drinkers: a practical class exercise in pharmacology. PMID- 9218934 TI - Pharmacokinetic interaction study of citalopram and cimetidine in healthy subjects. PMID- 9218936 TI - Bladder consequences of prostatic obstruction. AB - The development of new imaging techniques has improved our knowledge of the cross sectional anatomy of the pelvis and thereby our understanding of the bladder's response to prostatic obstruction. It is now known that changes in the bladder are intimately linked to the development of irritative and obstructive symptoms of BPH. This paper illustrates the anatomical changes within the pelvic region that can develop as a consequence of prostatic enlargement. The anatomical position of the prostate is such that its enlargement inevitably affects neighbouring organs. Enlargement of the median lobe can cause irritation of the trigone and the physical trapping of urine behind the lobe. The urethra may also become mechanically obstructed leading to an increase in the detrusor muscle mass as it struggles to overcome the restriction of the bladder outlet. In time, a change in the collagen to muscle ration in favour of collagen may develop, with the consequence that bladder compliance is reduced and the ability to increase volume without increasing pressure is diminished. Thereafter one of two scenarios may develop: the 'high pressure conflict' bladder or the 'distended low pressure' bladder. Urinary disturbance in BPH therefore results from an inadequate balance between bladder contractility and urethral resistance. PMID- 9218935 TI - Lack of significant stereoselectivity in pharmacokinetics of tiaprofenic acid. PMID- 9218937 TI - Modification of bladder structure in response to outflow obstruction and ageing. AB - The morphology of the normal bladder has been compared with that of the obstructed and the ageing bladder, using both light and electron microscopy. In the normal bladder the smooth muscle cells are closely packed together with relatively little intervening connective tissue. The smooth muscle bundles are innervated predominantly by autonomic presumptive cholinergic nerve fibres, while adrenergic nerves are usually observed in association with blood vessels. In the obstructed bladder, many smooth muscle cells are surrounded by large amounts of connective tissue and some of the muscle cells change their function from contracting to seemingly being involved in collagen synthesis. A significant reduction in the innervation of the smooth muscle cells is observed. In the ageing bladder, the smooth muscle cells have a normal morphology which is in contrast to the findings in the obstructed bladder. A reduction in the innervation of the ageing bladder is observed, although to a considerably lesser extent than that observed in the obstructed bladder. As the morphology of the obstructed bladder differs from that of the ageing bladder, it is unlikely that the changes seen in the obstructed bladder simply reflect a function of age. PMID- 9218938 TI - Cellular and molecular aspects of bladder hypertrophy. AB - Bladder dysfunction secondary to benign prostatic hyperplasia (BPH) is a major affliction associated with ageing. As the disease slowly progresses, the bladder changes from a state of compensation to decompensation, in which there are severe, irreversible alterations in bladder function. Using a rabbit model of partial outlet obstruction we have identified three major cellular changes in the bladder which result from such obstruction. These include progressive denervation, mitochondrial dysfunction and disturbances of calcium storage and release from the sarcoplasmic reticulum. Our hypothesis is that outlet obstruction results in bladder hypertrophy which induces ischaemia. This leads to a release of intracellular calcium, leading to activation of specific enzymes and generation of free radicals. These then attack the membranes of nerves, sarcoplasmic reticulum and mitochondria. We have demonstrated that pretreatment of rabbits with Pygeum africanum extract (Tadenan) significantly reduced the severity of both the contractile and metabolic dysfunctions induced by partial outlet obstruction. Our current hypothesis is that Tadenan may either prevent the activation of degradative enzymes (or generation of free radicals), or protect the intracellular membranes against the destructive effects of free radicals or degredative enzymes. In conclusion, identifying cellular mechanisms responsible for bladder dysfunction induced by partial outlet obstruction provides new possibilities for non-surgical treatment of BPH. Our studies on Tadenan support this concept that the bladder provides a novel target for therapeutic intervention. PMID- 9218939 TI - Role of growth factors in benign prostatic hyperplasia. AB - Benign prostatic hyperplasia (BPH) is a common condition among older men. Although surgery is the most effective treatment for patients with severe symptoms of bladder outlet obstruction, many patients with less severe symptoms will benefit from pharmacological intervention. Traditional medical therapies have involved hormonal manipulation. However, evidence has now emerged that prostate growth is under the immediate control of specific growth factors and only indirectly modulated by steroids. In this review we present a hypothesis of the mechanism of action of these growth factors in the developmental of BPH. Many of the more bothersome symptoms of BPH are not directly caused by the outlet obstruction, but only the resulting bladder hypertrophy. The development of the rabbit model of partial bladder outlet obstruction has allowed investigation of the bladder changes likely to occur in BPH. These studies have revealed the important role of growth factors in the development of bladder hypertrophy. Therefore, targeting growth factors potentially represents a direct therapeutic approach to the regulation of abnormal enlargement of the prostate and the amelioration of other symptoms associated with BPH. PMID- 9218940 TI - Clinical relevance of growth factor antagonists in the treatment of benign prostatic hyperplasia. AB - Changing demography and expectations about maintaining quality of life mean that an increasing number of men will require treatment for benign prostatic hyperplasia (BPH). Many growth factors have a role in the development of BPH. Consequently growth factor antagonists offer an attractive therapeutic option. In double-blind randomised trials Tadenan, a drug known to have growth factor antagonist activity, conferred significant improvement of urinary symptoms, maximum flow rate and residual volume, with no serious side-effects. Therapeutic outcome could be enhanced in a number of ways including matching patients with particular cell type overgrowth for treatment with a growth factor antagonist specific for that cell type. Full exploitation of this approach awaits the development of less invasive means of determining the cell type affected. PMID- 9218941 TI - Animal models of bladder outlet obstruction and molecular insights into the basis for the development of bladder dysfunction. AB - In humans, chronic bladder outlet obstruction resulting from benign prostatic hypertrophy (BPH) can induce severe and irreversible upper urinary tract changes, especially in the bladder. BPH can be mimicked in rabbits by artificially creating a partial obstruction of the bladder outlet. The structural and functional changes initiated eventually elicit a pathology and symptomology similar to human BPH. The rabbit bladder responds to partial outlet obstruction in three characteristic stages: hypertrophy, compensation and decompensation Basic fibroblast growth factor, epidermal growth factor and transforming growth factor-beta appear to be involved in the hypertrophy phase. During this initial phase normal bladder function is maintained but in the later phases, it is drastically impaired. We propose that the various stages of bladder remodelling subsequent to partial outlet obstruction are essentially driven by reduction in blood flow to the bladder (ischaemia). Initially this ischaemia might stimulate a compensatory response enabling the bladder to deal with the stress outlet obstruction. However, the long-term reduction in blood flow resulting from cyclical filling of the obstructed bladder, ultimately induces the degenerative changes that impede bladder function. These findings have important implications for the development of novel therapies to prevent or reverse bladder dysfunction resulting from BPH. PMID- 9218943 TI - Fetal rat brain injury: effect of transient maternal hypoxemia. AB - OBJECTIVE: To determine whether transient acute maternal hypoxemia during the end of pregnancy may cause neuronal damage in fetal rat brains. STUDY DESIGN: Nine pregnant rats (4 study and 5 controls) at 16-17 gestational days were studied. The study rats were placed in a chamber and breathed a gas mixture of 11.8% oxygen, 4.95% CO2, and nitrogen for either 1 or 2 h, while the control animals breathed room air. Tail venous blood was collected and gases were evaluated at the beginning and conclusion of the exposure periods. After 72 h of recovery, at 19-20 days' gestation, the fetal cardiovascular systems were perfused with saline and formalin. The brains were embedded in paraffin, sectioned in coronal plane, and stained with hematoxylin and eosin Histologic assessment of sections was performed by a neuropathologist blinded to the protocol. Statistical analysis of the data was performed using analysis of variance and the chi-square test. RESULTS: Exposure to the gas mixture resulted in decreased maternal pO2 from 39.9 +/- 7.6 mm Hg in the control group to 28.8 +/- 2.0 mm Hg in the 2-hour hypoxia group (p < 0.05). No significant changes in maternal pH or pCO2 status have been noted. A total of 34 fetal rat brains served as controls and 26 brains as the hypoxia study group There was a significant increase in isolated neuronal damage, including necrosis and shrinkage of cells, with karyorrhexis (fragmentation and breakage of the nucleus) in the hippocampus, basal ganglia, thalamus, and hypothalamus in the hypoxemia rats as compared with controls. CONCLUSION: Transient maternal hypoxemia may cause neuronal necrosis in vulnerable regions of the fetal rat brain, including the hippocampus, basal ganglia, thalamus, and hypothalamus. PMID- 9218942 TI - Nonimmune hydrops fetalis caused by intrauterine human parvovirus B19 infection: a case of spontaneous reversal in utero. AB - We report a case of spontaneous reversal in utero of hydrops fetalis caused by parvovirus B19 maternal-fetal infection. The route leading to fetal hydrops is not fully understood. Severe anemia with hypoxemia and viral fetal myocarditis have been incriminated. Then the main issue is fetal death or spontaneous abortion. Cases of spontaneous reversal of hydrops fetalis are unusual. Fetal regenerative anemia is a good prognostic factor and emphasizes the place of conservative management. PMID- 9218944 TI - Fetal breathing movements are associated with changes in compliance of the left ventricle. AB - We have tested the hypothesis that the limited fetal ventricular distensibility is not only an intrinsic cardiac characteristic but is also contributed to by the pressure exerted by the intrathoracic organs. To this purpose we have studied the diastolic cardiac function by Doppler velocimetry in 11 fetuses during fetal breathing and in 22 fetuses during apnea, controlling for gestational age at examination and heart rate. Inspiration was associated a significant increase in left ventricular compliance, as measured by deceleration time (inspiration 182.6 +/- 15.5 s vs. expiration 137.1 +/- 13.1 s vs. apnea 151.7 +/- 51.8 s), and in ventricular filling, as measured by velocity time integral (inspiration 0.086 +/- 0.020 m vs. expiration 0.064 +/- 0.014 m vs. apnea 0.065 +/- 0.011 m), compared with both the expiration and apnea groups. These increases most likely reflect changes in venous return and ventricular end-diastolic volume secondary to a decrease in intrathoracic pressure during fetal breathing. PMID- 9218945 TI - Effect of endoscopic white light on the developing rat retina. AB - OBJECTIVE: Our goal was determine the effect of endoscopic white light on the developing mammalian retina by quantitative histological analysis. STUDY DESIGN: Albino rats at postnatal days 10 (n = 16) and 16 (n = 20) were exposed to endoscopic white light for 1 h, while an equal number of littermate controls were not exposed, but otherwise treated identically. The thickness of the outer nuclear layer (ONL) and the total retinal thickness were measured light microscopically using a computer-assisted morphometry program. RESULTS: We found no statistically significant decrease in the thickness of the ONL or the total retinal thickness, nor were any significant decreases found in the ONL or the total retinal thickness for male or female groups. CONCLUSION: Endoscopic white light does not cause damage to the developing rodent neural retina detectable at the light microscopic level. PMID- 9218946 TI - Chorioamniotic membrane separation: a potentially lethal finding. AB - Sonographic detection of chorioamniotic membrane separation (CMS) has been considered a benign incidental finding. We now report 6 cases of CMS identified by prenatal ultrasound; 1 in an otherwise normal pregnancy and 5 following fetal surgery. Following membrane separation, amniotic bands formed and compromised the umbilical cord in 4 cases leading to 2 fetal deaths. In the first case, CMS was detected by ultrasound at 22 weeks' gestation in an otherwise uncomplicated pregnancy. Because CMS was considered benign and umbilical cord blood flow was ample, the mother was followed by intermittent sonographic examinations. Fetal demise occurred 2 weeks later, clearly due to umbilical cord strangulation by an amniotic band. Surprised by this unexpected outcome, we reviewed our experience with CMS after hysterotomy for fetal surgery. Out of more than 40 fetal surgical cases, we have 5 cases in which CMS was recognized after hysterotomy. Three of these fetuses had umbilical cord compromise by a band of amniotic membrane leading to 1 fetal death. This experience demonstrates that membrane separation may be associated with amniotic band formation which can lead to cord strangulation and fetal compromise. Following fetal surgery, serial ultrasound evaluation and close fetal monitoring are indicated. In otherwise unremarkable pregnancies, clinician awareness of the possibility of amniotic band formation following CMS should be heightened. In either situation, knowledge of this potential life-threatening complication may identify cases in which cord compromise requires emergent delivery or fetoscopic release of the strangulating amniotic band. PMID- 9218947 TI - Doppler velocimetry of the umbilical artery as a predictor of pregnancy outcome in pregnancies characterized by elevated maternal serum alpha-fetoprotein and normal amniotic fluid alpha-fetoprotein. AB - OBJECTIVE: Women with elevated maternal serum alpha-fetoprotein (MSAFP) and normal amniotic fluid alpha-fetoprotein (AFAFP) are at an increased risk of an adverse pregnancy outcome. Such MSAFP elevations are probably the consequence of transplacental leakage caused by placental abnormalities. These may result in disturbed bloodflow through placental vessels. The purpose of this study was to assess whether measurement of such disturbances by Doppler velocimetry of the umbilical artery has a predictive value for pregnancy outcome. STUDY DESIGN: The study group consisted of 85 patients, in whom the only finding was elevated maternal serum alpha-fetoprotein. Systolic/diastolic (S/D) ratios were calculated using a continuous wave Doppler measurement of the umbilical artery, performed at 6 to 8-week intervals. Serial results for each individual were incorporated into a single 'Velocimetry Score'. RESULTS: In group B (14 patients) with an abnormally elevated umbilical S/D ratio, a higher incidence of intrauterine growth retardation (42.9%), preterm deliveries (78.6%), and fetal loss (42.9%) was noted, as compared with group A (71 patients) with a normal S/D ratio. CONCLUSIONS: Umbilical artery Doppler velocimetry may serve as a predictor of pregnancy outcome in the high-risk group characterized by elevated MSAFP. PMID- 9218948 TI - Erythromycin treatment for subclinical Ureaplasma urealyticum infection in preterm labor. AB - This study was undertaken to test the effects of erythromycin as an adjunct to tocolysis for preterm labor in women with vaginal cultures positive for Ureaplasma urealyticum. The study group consisted of 18 women in active preterm labor with pregnancies between 26 and 34 weeks of gestation and intact membranes who received 500 mg erythromycin orally every 8 h for 10 days. Seventeen women with similar characteristics served as controls and received no antibiotics. In all women contractions were suppressed with ritodrine. Erythromycin treatment resulted in a statistically significant greater mean delay of delivery (36.4 days) than among the control group (23.1 days). Higher proportion of term pregnancies (7 versus 3 pregnancies), higher mean birth weight (2,745 versus 2,474 g), lower neonatal morbidity (22.2 versus 42.2%) and shorter mean neonatal hospitalization time (9.6 versus 12.1 days) were observed, although these differences were not statistically significant. Adjunctive erythromycin treatment given to women treated for preterm labor with intact membranes and positive vaginal cultures for U. urealyticum appears to prolong gestation and to improve perinatal outcome. PMID- 9218949 TI - Results and views of women in population-wide pregnancy screening for trisomy 21 in east Finland. AB - The serum screening test for trisomy 21 was offered to all pregnant women in East Finland during 1993. The results and the women's views on trisomy screening are reported here. A total of 9,343 (89%) of the pregnant women participated in the screening. A total of 615 women were eligible for amniocentesis or chorionic villus sampling; among them 22 pregnancies were affected by Down Syndrome. Fifteen of them were detected by double screening; in 2 missed cases only maternal-serum alpha-fetoprotein had been determined. In all 5 cases missed by double screening the maternal age was less than 30 years. The women's attitudes were studied by means of a questionnaire, which was mailed to 214 women. The response rate to our questionnaire was 74%. A total of 87% of respondents highly or very highly valued the opportunity to participate in the screening. Despite the existing problems caused by late testing, 87% of respondents want to be tested again in their future pregnancies. PMID- 9218950 TI - The early amniocentesis study: a randomized clinical trial of early amniocentesis and midtrimester amniocentesis. II. Evaluation of procedure details and neonatal congenital anomalies. AB - The present study provides detailed neonatal and congenital malformation follow up from 695 women enrolled in a prospective randomized multicenter study comparing the safety and accuracy of early (11-12 weeks of gestation) and midtrimester amniocentesis (15-16 weeks of gestation). No differences were found for total pregnancy loss (difference 0.4%; CI -3.6 to 4.4%), obstetrical or neonatal outcome. The incidence of congenital anomalies was 2.4 and 2.6% for the early amniocentesis and midtrimester amniocentesis groups, respectively. Respiratory problems were present in 2.1 and 1.6%, respectively. Musculoskeletal problems were present in 0.9 and 2.4%, respectively. It must be emphasized that before early amniocentesis can be considered as an alternative to midtrimester amniocentesis or chorionic villus sampling, careful evaluation of any fetal effects must be considered and further large randomized trials are necessary. PMID- 9218952 TI - Coiled-coil assembly by peptides with non-heptad sequence motifs. AB - BACKGROUND: The seven-residue heptad repeat is the accepted hallmark of coiled coils. In extended filamentous proteins, however, contiguous patterns of heptads are often disrupted by 'skips' and 'stammers'. The structural consequences and roles of these digressions are not understood. RESULTS: In a cytoskeleton protein from Giardia lamblia, heptads flank eleven-residue units (hendecads) to give a 7 11-7 motif that dominates the sequence. Synthetic peptides made to the consensus sequence of this motif fold in solution to fully helical, parallel dimers. Both the sequence pattern and these experimental data are consistent with the coiled coil model. We note that breaks in other extended coiled coils can also be reconciled by hendecad insertions. CONCLUSIONS: The heptad paradigm for the coiled coil must be expanded to include hendecads. As different combinations of heptads and hendecads will give different overall sequence motifs, we propose that these provide a mechanism to promote cognate protein pairings during the folding of extended coiled coils in the cell. PMID- 9218951 TI - Effect of Hispanic ethnicity on interpretation of maternal serum screening. AB - Maternal serum concentrations of alpha-fetoprotein (MSAFP), unconjugated estriol (uE3) and human chorionic gonadotropin (hCG) were measured in the sera of 3,046 Hispanic women and of 15,154 Caucasian women from gestational weeks 14 through 20 between January 1990 and December 1995. Median values for analytes were calculated for each gestational week and the two ethnic groups compared. Our findings indicate that the median values for Hispanics are lower for MSAFP and hCG, but higher for estriol. These differences are not accounted for by differences in median weights of the two ethnic groups. Although these results would suggest that Hispanic women at risk for a fetus with a neural tube defect (NTD) may be missed when cut-offs derived from a Caucasian population are used, in fact, that was not the case in our screening population. However, a disproportionate number of Hispanic women may be identified with an abnormal serum screening test unrelated to a risk for neural tube defects. This does not infer that detection rates are different across different ethnic groups since other factors are involved. Adjustment of medians for Hispanic ethnicity may have a small but significant effect, especially with regard to low values. PMID- 9218953 TI - The future of protein secondary structure prediction accuracy. AB - BACKGROUND: The accuracy of secondary structure prediction for a protein from knowledge of its sequence has been significantly improved by about 7% to the 70 75% range by inclusion of information residing in sequences similar to the query sequence. The scientific literature has been inconsistent, if not negative, regarding chances for further improvement from the vast knowledge to be provided by genome sequencing efforts. RESULTS: By applying a prediction technique that is particularly sensitive to added sequence information to a standard set of query sequences with related primary structures taken from chronologically successive releases of the SWISS-PROT database, it is shown that prediction accuracy can be expected to reach 80-85% with a large 10-fold increase in present sequence knowledge. CONCLUSIONS: Even with present prediction approaches, improvement in prediction accuracy can still be expected, albeit limited to no more than 10%. PMID- 9218954 TI - Conformational transitions provoked by organic solvents in beta-lactoglobulin: can a molten globule like intermediate be induced by the decrease in dielectric constant? AB - BACKGROUND: It is known that nonnative states of protein molecules can exist in living cells and can be involved in a number of physiological processes. It has also been established that the membrane surface can be responsible for the partial denaturation of proteins due to negative charges on it. The local decrease in the effective dielectric constant of water near the organic surface has been suggested to be an additional driving force for protein denaturation in the membrane field, but data to confirm this suggestion were lacking. RESULTS: Conformational transitions induced in beta-lactoglobulin by methanol, ethanol, isopropanol, dimethylformamide and dioxane were studied by near and far UV circular dichroism, steady-state tryptophan fluorescence and fluorescence decay of 8-anilinonaphthalene-1-sulfonate (8-ANS). The existence of at least two noncoinciding cooperative transitions has been established in all solvent systems studied. The first of these transitions describes the disruption of rigid tertiary structure in protein molecules, while the second reflects the formation of an expanded helical conformation typical of proteins in concentrated organic solvents. This means that the organic solvents provoke the formation of a denatured intermediate state with pronounced secondary structure and native-like compactness. We show that the positions of maxima in fI versus dielectric constant dependence virtually coincide for all five solvent systems studied. CONCLUSIONS: The decrease in the dielectric constant of the solvent induces in beta-lactoglobulin an equilibrium intermediate state. This state, being denatured, is relatively compact and has pronounced secondary structure and high affinity for the hydrophobic fluorescent probe 8-ANS, i.e. possesses all the properties of the molten globule intermediate state. PMID- 9218955 TI - Direct evaluation of thermal fluctuations in proteins using a single-parameter harmonic potential. AB - BACKGROUND: An elastic network model is proposed for the interactions between closely (< or = 7.0 A) located alpha-carbon pairs in folded proteins. A single parameter harmonic potential is adopted for the fluctuations of residues about their mean positions in the crystal structure. The model is based on writing the Kirchhoff adjacency matrix for a protein defining the proximity of residues in space. The elements of the inverse of the Kirchhoff matrix give directly the auto correlations or cross-correlations of atomic fluctuations. RESULTS: The temperature factors of the C alpha atoms of 12 X-ray structures, ranging from a 41 residue subunit to a 633 residue dimer, are accurately predicted. Cross correlations are also efficiently characterized, in close agreement with results obtained with a normal mode analysis coupled with energy minimization. CONCLUSIONS: The simple model and method proposed here provide a satisfactory description of the correlations between atomic fluctuations. Furthermore, this is achieved within computation times at least one order of magnitude shorter than commonly used molecular approaches. PMID- 9218956 TI - Homology modelling of an antimicrobial protein, Ace-AMP1, from lipid transfer protein structures. AB - BACKGROUND: Plant nonspecific lipid transfer proteins (ns-LTPs) are small basic proteins that facilitate lipid shuttling between membranes in vitro. The function of ns-LTPs in vivo is still unknown. It has been suggested, in relation to their lipid binding ability, that they may be involved in cutin formation. Alternatively, they may act in the plant defence system against pathogenic agents. Ace-AMP1 is an antimicrobial protein extracted from onion seed that shows sequence homology with ns-LTPs but that is unable to transfer lipids. We have recently determined the three-dimensional structure of wheat and maize ns-LTPs. In order to compare the structural features of Ace-AMP1 and ns-LTPs, we have used the comparative modelling software MODELLER to predict the structure of Ace-AMP1. RESULTS: The global fold of Ace-AMP1 is very similar to those of ns-LTPs, involving four helices and a C-terminal tail without secondary structure elements. The structure of maize and wheat ns-LTP is characterized by the existence of a tunnel-like hydrophobic cavity in which a lipid molecule can be inserted. In the Ace-AMP1 structure, this cavity is blocked by a number of bulky residues. Similarly, the electrostatic potential contours of ns-LTPs show some common features that were not observed in Ace-AMP1. CONCLUSIONS: Although Ace AMP1 displays a similar global fold to ns-LTPs, it does not present a hydrophobic cavity, which may explain why Ace-AMP1 cannot shuttle lipids between membranes in vitro. The large differences in the electrostatic properties of Ace-AMP1 and ns LTPs suggest a different mode of interaction with membranes. PMID- 9218958 TI - Distance geometry based comparative modelling. AB - A distance geometry based protein modelling algorithm is presented which relies on the projection of simple model chain coordinates into Euclidean spaces with gradually decreasing dimensionality. Fast embedding was achieved by performing separate distance matrix projections on subsets of the model points. Structural equivalences between the unknown target and related proteins with known structures were deduced either from a mixed sequence/structure multiple alignment or from the output of various fold recognition (threading) approaches. These equivalences were mapped onto the model as structure-specific conserved C alpha atom distances and secondary structure assignments. Additional nonspecific distance restraints derived from general stereochemical properties of folded protein chains were used to guide the modelling process. The method quickly constructed a large number of low-resolution models which could then serve as starting conformations for full-atom refinement. Structure predictions for some targets in the 'Asilomar Challenge' (CASP2) are presented to illustrate potential applications of the approach. PMID- 9218957 TI - Geometrical factor and physical reasons for its influence on the kinetic and thermodynamic properties of RNA-like heteropolymers. AB - BACKGROUND: It has been shown that not only the primary sequence but also the geometry of the native conformation influences the kinetic and thermodynamic properties of a protein-like model. The purpose of this paper is to elucidate the geometrical factor, which affects the kinetic and thermodynamic properties of RNA like heteropolymers, and physical reasons for this phenomenon. RESULTS: It is shown that increasing the strength of long-range contacts can accelerate the finding of the native secondary structure for RNA-like heteropolymers. Decreasing the strength of long-range contacts results in deceleration of this process. The physical reason for this phenomenon is the increase in the average energy of non native structures and, as a consequence, the increase (in absolute value) of the relative value of native energy from strengthening long-range contacts under constant energy of the native state. Statistical analysis of natural RNAs has shown that the mean stability of helices formed by the pairing of regions that are remote along the chain prevails over the mean stability of helices formed by regions that are close along the chain. CONCLUSIONS: The kinetics simulations suggest that the folding of the RNA secondary structure depends on the structural factors of the native state. One of these factors accelerating large RNA folding is the existence of strong long-range helices in the native secondary structure. PMID- 9218960 TI - Extraction of well-fitting substructures: root-mean-square deviation and the difference distance matrix. AB - The extraction of well-fitting substructures of two or more sets of proteins has applications to analysis of mechanisms of conformational change in proteins, including pathways of evolution, to classification of protein folding patterns, and to evaluation of protein structure predictions. Many methods are known for extracting some substantial common substructure with low root-mean-square deviation (r.m.s.d.). A harder problem is addressed here: finding all common substructures with r.m.s.d. less than a prespecified threshold. Our approach is to consider the minimum value of the maximum distance between corresponding points, corresponding to superposition in the Chebyshev norm. Using the properties of Chebyshev superposition, we derive relationships between the r.m.s.d. and the maximum element of the difference matrix, two common measures of structural similarity. The results provide a basis for developing algorithms and software to identify all well-fitting subsets. PMID- 9218959 TI - Macromolecular mimicry in protein biosynthesis. AB - Elongation factor Tu (EF-Tu) is a G-protein which, in its active GTP conformation, protects and carries aminoacylated tRNAs (aa-tRNAs) to the ribosome during protein biosynthesis. EF-Tu consists of three structural domains of which the N-terminal domain consists of two special regions (switch I and switch II) which are structurally dependent on the type of the bound nucleotide. Structural studies of the complete functional cycle of EF-Tu reveal that it undergoes rather spectacular conformational changes when activated from the EF-Tu.GDP form to the EF-Tu.GTP form. In its active form, EF-Tu.GTP without much further structural change interacts with aa-tRNAs in the so-called ternary complex. The conformational changes of EF-Tu involve rearrangements of the secondary structures of both the switch I and switch II regions. As the switch II region forms part of the interface between domains 1 and 3, its structural rearrangement results in a very large change of the position of domain 1 relative to domains 2 and 3. The overall shape of the ternary complex is surprisingly similar to the overall shape of elongation factor G (EF-G). Thus, three domains of the protein EF-G seem to mimic the tRNA part of the ternary complex. This macromolecular mimicry has profound implications for the function of the elongation factors on the ribosome. PMID- 9218961 TI - Coherent topological phenomena in protein folding. AB - A theory is presented for coherent topological phenomena in protein dynamics with implications for protein folding and stability. We discuss the relationship to the writhing number used in knot diagrams of DNA. The winding state defines a long-range order along the backbone of a protein with long-range excitations, 'wring' modes, that play an important role in protein denaturation and stability. Energy can be pumped into these excitations, either thermally or by an external force. PMID- 9218962 TI - Protein structures sustain evolutionary drift. AB - A protein sequence folds into a unique three-dimensional protein structure. Different sequences, though, can fold into similar structures. How stable is a protein structure with respect to sequence changes? What percentage of the sequence is 'anchor' residues, that is, residues crucial for protein structure and function? Here, answers to these questions are pursued by analyzing large numbers of structurally homologous protein pairs. Most pairs of similar structures have sequence identity as low as expected from randomly related sequences (8-9%). On average, only 3-4% of all residues are 'anchor' residues. The symmetric shape of the distribution at low sequence identity suggests that for most structures, four billion years of evolution was sufficient to reach an equilibrium. The mean identities for convergent (different ancestor) and divergent (same ancestor) evolution of proteins to similar structures are quite close and hence, in most cases, it is difficult to distinguish between the two effects. In particular, low levels of sequence identity appear not to be indicative of convergent evolution. PMID- 9218963 TI - Improving contact predictions by the combination of correlated mutations and other sources of sequence information. AB - We have previously developed a method for predicting interresidue contacts using information about correlated mutations in multiple sequence alignments. The predictions generated with this method were clearly better than random but not enough for their use in de novo protein folding experiments. We assess the possibility of improving contact predictions combining information from the following variables: correlated mutations, sequence conservation, sequence separation along the chain, alignment stability, family size, residue-specific contact occupancy and formation of contact networks. The application of a protocol for combining these independent variables leads to contact predictions that are on average two times better than those obtained initially with correlated mutations. Correlated mutations can be effectively combined with other types of information derived from multiple sequence alignments. Among the different variables tried, sequence conservation and contact density are particularly relevant for the combination with correlated mutations. PMID- 9218964 TI - Nocturnal partial seizures and arousals/awakenings from sleep: an ambulatory EEG study. AB - The architecture of nocturnal sleep in twenty subjects (9 males, 11 females, mean age 27 years) affected by partial cryptogenic epilepsy was investigated by means of ambulatory EEG (A-EEG) recording performed at home. The study aimed in particular to ascertain the immediate effects of nocturnal partial epileptic seizures on sleep stability and continuity. Data for a total of 49 recorded seizures indicate that 72% of the partial seizures which occurred during sleep were followed by arousal and awakening, and that sleep interruption lasted significantly longer when the seizure occurred between 4 and 7 a.m. PMID- 9218965 TI - Cardiac autonomic involvement and peripheral nerve function in patients with diabetic neuropathy. AB - The aim of our study was to investigate the relationship between cardiac autonomic neuropathy and dysfunction of myelinated and unmyelinated nerve fibres in the peripheral nerve. We measured nerve conduction velocities, warmth/cold perception thresholds at the foot dorsum, sympathetic skin response (SSR), and performed the quantitative sudomotor axon reflex test (QSART). Forty-three diabetic patients with distal-symmetric polyneuropathy were included. According to the results of heart rate variation, 20 patients had cardiac autonomic neuropathy (CAN+). Apart from motor nerve conduction velocities, all tests were more often abnormal in CAN+ patients. Warmth thresholds (afferent C-fibres) and reduced compound muscle action potentials (CMAPs) of the tibial and peroneal nerve, indicating axonal damage, were more often abnormal in CAN+. Cold threshold and sural nerve conduction velocity were indicators of involvement of myelinated small and large nerve fibres, but not of the cardiac autonomic system. Ninety four percent (94%) of patients with absent SSR and 78% of patients with abnormal QSART had CAN+. SSR and QSART may be useful for assessment of autonomic neuropathy in diabetic patients with cardiac arrhythmia where direct measurement of heart rate variability is not possible. In the majority of our patients with CAN+, the vagal-cardiac and the sudomotor-sympathetic systems were involved simultaneously, although two entirely different systems were tested. This may reflect a C-fibre directed selectivity of the pathological process in autonomic diabetic neuropathy. In conclusion our results show that diabetics with and without cardiac autonomic neuropathy have a different profile of involvement of peripheral nerve fibres. PMID- 9218966 TI - Congenital (?) Horners syndrome and ipsilateral headache. AB - A 31-year-old woman had left-sided miosis, ptosis, and hypopigmented iris probably since birth. At 22, she developed intermittent headaches, always in the left frontotemporal region. These headaches lasted from 1 to 2 days and recurred every 1-2 months. Pain attacks were pressing-pulsatile in character, moderate in intensity, and frequently accompanied by nausea, vomiting, and moderate phono- and photophobia. Various treatment alternatives, such as conventional analgesics and ergotamine failed to improve the attacks. Pizotifen was partially effective. The results of pupillometry and evaporimetry studies were consistent with a 3rd neuron sympathetic hypofunction on the symptomatic side. Autonomic studies and clinical features were consistent with a congenital Horner's syndrome. Conceivably, a sympathetic hypofunction may play a role in the pathogenesis of such headache or in its lateralization. Indomethacin and sumatriptan both seemed to provide absolute pain relief. Some clinical features, the fact that the IHS criteria for migraine are fulfilled and that sumatriptan is efficient, demonstrate the similarity to migraine. The coexistence of strict unilaterality of pain and the probable, complete response to indomethacin indicate a similarity to hemicrania continua in its remitting form. Further information on the effect of sumatriptan in hemicrania continua will help clarify the position of this case vs. hemicrania continua. At this stage, it is probably not possible to classify this case properly. PMID- 9218967 TI - "Ottorino Rossi" Award 1997. The biology of nitric oxide. PMID- 9218968 TI - Alzheimer's disease: phenotypes and genotypes. PMID- 9218969 TI - The cholinergic system in Alzheimer's disease and dementia with Lewy bodies: from animal models to neuropathological data. PMID- 9218970 TI - Creutzfeldt-Jakob disease and related disorders: etiopathogenetic aspects. PMID- 9218971 TI - Clinical neuropathology of Creutzfeldt-Jakob disease. PMID- 9218972 TI - Glucocorticoid hormone, aging brain and dementia. PMID- 9218973 TI - Cognitive function at menopause: neuroendocrine implications for the study of the aging brain. PMID- 9218974 TI - Neuropeptides and senile dementia: focus on galanin. PMID- 9218975 TI - Manipulation of the cholinergic system. PMID- 9218976 TI - Noradrenaline loss selectivity exacerbates nigrostriatal toxicity in different species of rodents. PMID- 9218977 TI - The role of astroglial cell-derived nitric oxide and prostanoids in neurodegenerative disorders. PMID- 9218978 TI - Somatic pathology in elderly demented patients. PMID- 9218979 TI - Behavioral disorders in dementia. PMID- 9218980 TI - Pharmaceutical treatment of cognitive disorders in Alzheimer's disease. PMID- 9218981 TI - Comparison of the memory performances of young and old subjects with those of Alzheimer's disease patients: support for a discontinuity between natural and pathological aging. PMID- 9218982 TI - Non pharmacological treatment in Alzheimer's disease. PMID- 9218983 TI - Medium-term outcomes of Alzheimer patients in special care units. PMID- 9218984 TI - Treatment of impaired cognition with nootropic drugs: nicergoline versus the state of the art. PMID- 9218985 TI - Population-based care of depression: effective disease management strategies to decrease prevalence. AB - This paper reviews the concepts of population-based care and disease management of major depression. Population-based care and disease management strategies motivated by health care reform provide approaches for organizing health services to lower the prevalence of common medical and psychiatric illnesses in primary care populations. We apply these concepts to the organization of services for patients with major depression. PMID- 9218986 TI - Recognition and treatment of depressive disorders by internal medicine attendings and housestaff. AB - Depression is underdiagnosed and undertreated by nonpsychiatric practitioners. Research suggests improvement is needed in the recognition and treatment of depressive disorders by primary care physicians. This study was undertaken to better understand internists' ability to recognize depressive disorders, choice of appropriate medications, dosage, and treatment patterns. Questionnaires were distributed to 45 internal medicine attendings, 45 internal medicine housestaff, and 32 adult psychiatry residents. Each questionnaire contained four vignettes: major depressive disorder (MDD), MDD with melancholic features, MDD with atypical features, and MDD with psychotic features. Eleven questions per case covered diagnoses, management, and treatment. Data analysis with intragroup comparisons on 20 internal medicine attendings, 33 internal medicine housestaff, and 32 psychiatry residents suggested that many internal medicine attendings and housestaff had difficulty in recognizing major depression and its subtypes. Although the findings indicated that internists would initiate pharmacological treatment, they frequently made incorrect or questionable pharmacological choices. Psychiatric referral or consultation was often endorsed. Our findings among internists are consistent with previous research examining other primary care physicians suggesting that depression is underdiagnosed and undertreated. PMID- 9218987 TI - Chronic fatigue syndrome. A practical guide to assessment and management. AB - Chronic fatigue and chronic fatigue syndrome (CFS) have become increasingly recognized as a common clinical problem, yet one that physicians often find difficult to manage. In this review we suggest a practical, pragmatic, evidence based approach to the assessment and initial management of the patient whose presentation suggests this diagnosis. The basic principles are simple and for each aspect of management we point out both potential pitfalls and strategies to overcome them. The first, and most important task is to develop mutual trust and collaboration. The second is to complete an adequate assessment, the aim of which is either to make a diagnosis of CFS or to identify an alternative cause for the patient's symptoms. The history is most important and should include a detailed account of the symptoms, the associated disability, the choice of coping strategies, and importantly, the patient's own understanding of his/her illness. The assessment of possible comorbid psychiatric disorders such as depression or anxiety is mandatory. When the physician is satisfied that no alternative physical or psychiatric disorder can be found to explain symptoms, we suggest that a firm and positive diagnosis of CFS be made. The treatment of CFS requires that the patient is given a positive explanation of the cause of his symptoms, emphasizing the distinction among factors that may have predisposed them to develop the illness (lifestyle, work stress, personality), triggered the illness (viral infection, life events) and perpetuated the illness (cerebral dysfunction, sleep disorder, depression, inconsistent activity, and misunderstanding of the illness and fear of making it worse). Interventions are then aimed to overcoming these illness-perpetuating factors. The role of antidepressants remains uncertain but may be tried on a pragmatic basis. Other medications should be avoided. The only treatment strategies of proven efficacy are cognitive behavioral ones. The most important starting point is to promote a consistent pattern of activity, rest, and sleep, followed by a gradual return to normal activity; ongoing review of any 'catastrophic' misinterpretation of symptoms and the problem solving of current life difficulties. We regard chronic fatigue syndrome as important not only because it represents potentially treatable disability and suffering but also because it provides an example for the positive management of medically unexplained illness in general. PMID- 9218988 TI - Ethical and legal implications of managed care. AB - This article addresses several ethical, regulatory, and legal issues in managed care with attention to recent court cases that focus on physicians' responsibility, fiduciary duty, and the impact that these legal decisions have on physicians practicing in a managed care environment. Discussion of the impact of changes in the control of decision-making processes for physicians, the use of managed care protocols, restriction of resources, and gatekeeping systems are addressed as are the specific duties and obligations of physicians to their patients. PMID- 9218989 TI - Serious overdosers admitted to a general hospital: comparison with nonoverdose self-injuries and medically ill patients with suicidal ideation. AB - There are few psychiatric studies of serious self-injury patients admitted to general hospitals. In order to better characterize patients whose overdoses are serious enough to require hospitalization on a toxicology service, we describe 207 consecutively admitted, serious overdose patients (OD), all of whom were psychiatrically evaluated. They were compared with 53 nonoverdose self-injury cases (NO) and 79 medical/surgical patients with suicidal ideation (SI) who were routinely referred for consultation during the same 2-year period. All data were contemporaneously compiled into a computerized database and analyzed. The attempters (OD and NO) were younger than the ideation patients (SI). The OD group was predominantly female (60%) and the NO and SI groups were predominantly male (72%). More OD cases were separated and more SI cases were widowed. Similar to previous reports, prior psychiatric contact was high in all groups. DSM-III-R diagnoses of depression, adjustment disorders, and substance abuse were most common in each group, without group differences. Only borderline personality disorder distinguished the groups, and the OD group had significantly more borderline patients. There were no seasonal differences for admission dates between groups, SI cases had longer hospital lengths of stay and were least likely to require further psychiatric care after discharge from the general hospital. Attempter groups were more similar to each other than the ideation patients. Clinicians should maintain a high index of suspicion for Axis I disorders in suicidal general hospital patients, though female borderline patients are particularly associated with the serious overdose method. PMID- 9218990 TI - Geropsychiatric consultation for African-American and Caucasian patients. AB - We assessed differences in the referral rates of African-American and Caucasian elderly for geropsychiatric consultation. Reasons for referral, assigned diagnoses, and interventions made were also compared. A retrospective chart review of psychiatric consultations was completed for patients aged 65 years and older for a 2-year period. Significantly more consultations were requested Caucasian elderly (6.2%) than for African-American elderly (3.8%). African American elderly were referred for evaluation of psychosis significantly more often and for assessment of suicide potential significantly less often than Caucasians. Consultants diagnosed African-American elderly with psychotic disorders, specifically schizophrenia, and dementia significantly more often than Caucasians. Caucasian elderly were significantly more often diagnosed with mood disorders, especially depressive disorders. Interventions/recommendations made for Caucasian and African-American elderly did not differ for the most part. Recommendations for legal measures were suggested for African-American elderly more often than for Caucasians. Differences between Caucasian and African American elderly were observed in consultation referral rates, reasons for referral, and psychiatric diagnoses made. The potential impact of cultural variables and the racial and age differences between hospital staff and patients may account for some of these findings. Further awareness of the needs of African American elderly is required. PMID- 9218991 TI - Psychosis in medical conditions: response to risperidone. AB - We report the response to risperidone in seven hospitalized, adult patients who presented psychotic symptoms etiologically related to a general medical condition. The conditions included brain surgery in two, and anticardiolipin syndrome, renal failure, epilepsy, lupus, and metastatic carcinoma in one each. Four patients had failed previous treatment with at least one typical antipsychotic agent. Response to risperidone was assessed by the Brief Psychiatric Rating Scale (BPRS). Serum was collected for measurement of steady state trough risperidone and 9-hydroxyrisperidone concentrations at effective doses in three patients. Amelioration of psychotic symptoms was noted in all seven patients. Mean (+/- SD) BPRS scores were reduced significantly from baseline (63.0 +/- 15.1) to endpoint (27.0 +/- 3.5; p < 0.01). The mean effective daily dose of risperidone was 3.1 +/- .7 mg and time to response was 4.7 +/- 2.4 days. Risperidone was not present at detectable concentrations in the three patients studied. The mean steady-state trough serum concentration of 9 hydroxyrisperidone in the three patients assessed was 20.3 +/- 9.8 ng/ml. These preliminary findings, which suggest that risperidone is a safe and effective agent in patients with psychotic symptoms due to various medical conditions, need to be confirmed by randomized, antipsychotic comparison trials involving a larger number of patients. PMID- 9218992 TI - Alexithymic features do not predict compliance with psychotherapy in consultation liaison patients. AB - A 1-year follow-up study on 54 general hospital psychiatric consultation outpatients was carried out in order to determine whether alexithymic features, measured by the Toronto Alexithymia Scale (TAS) are predictive of psychotherapy recommendations and whether alexithymia is associated with patients' compliance with these recommendations. Contrary to what we expected, the presence of alexithymic features predicted neither treatment recommendations nor compliance. Psychological distress as measured by the Brief Symptom Inventory (BSI) proved to be a better predictor. PMID- 9218993 TI - Mean age of tumor onset in hereditary nonpolyposis colorectal cancer (HNPCC) families correlates with the presence of mutations in DNA mismatch repair genes. AB - Fourteen Italian families affected with hereditary nonpolyposis colorectal cancer (HNPCC) were screened for germline mutations at three DNA mismatch repair (MMR) genes, MSH2, MLHI, and GTBP, by using a combination of different methods that included an in vitro synthesized protein assay, single-strand conformation polymorphism analysis, and direct sequencing. DNA alterations were observed in six instances, including a single base deletion in MSH2 exon 14, an A-to-G transition in the splice donor site of MLHI exon 6, and two missense mutations in MLHI exons 5 and 9. A previously reported common mutation affecting the splice donor site of MSH2 exon 5 was identified in two families. No mutations were detected in the GTBP gene. In total, eight of 16 Italian HNPCC families (50%), including two previously reported kindreds, were found to carry a mutation in MMR genes. We compared the mean age of colorectal cancer onset in the index cases (three patients for each family) between the two groups of kindreds, those with identified mutation vs. those without, and found that the first had a significantly lower value (43.0 vs. 53.7 years, P = 0.014). This finding suggests that HNPCC families with a more advanced age of tumor onset are less likely to be associated with known MMR genes. PMID- 9218994 TI - Promiscuous translocations of chromosome arm 17q in human neuroblastomas. AB - Deletions of chromosome arm 1p and amplification of the MYCN oncogene are well recognized genetic changes in neuroblastoma cells. Technical difficulties in cytogenetic analysis of this tumour have hampered the recognition of other recurring abnormalities, but recent use of molecular cytogenetic techniques has indicated significant involvement of chromosome arm 17q. In primary tumours and in cell lines, a recurrent unbalanced translocation t(1p;17q) has been identified by fluorescence in situ hybridization. We confirm the occurrence of this translocation in primary tumours and, in addition, we describe seven new structural rearrangements all of which result in gain of 17q in tumour cells. These rearrangements involved chromosome arms 9p, 10q, 11p, 14q, and 16q. Triplication of the 17q arm was seen in one case. The 17q breakpoint was most commonly q21. All these 17q changes were found in near-diploid tumours. We have also reviewed the literature for neuroblastoma karyotypes involving 17q abnormalities; taken in conjunction with our findings this indicates a remarkable promiscuity of translocation partners, with more than 20 different chromosome regions involved in 17q translocations. PMID- 9218995 TI - Inversion of chromosome 11 inv(11)(p15q22), as a recurring chromosomal aberration associated with de novo and secondary myeloid malignancies: identification of a P1 clone spanning the 11q22 breakpoint. AB - We studied four patients with inv(11)(p15q22) associated with malignant myeloid diseases by using fluorescence in situ hybridization (FISH) with phage and cosmid probes mapped and ordered on 11q22-24. Two of the four patients had non-Hodgkin's lymphoma or acute lymphoblastic leukemia as the primary malignancy and had received cytotoxic chemotherapy, including topoisomerase II inhibitors. The other two had de novo acute myeloid leukemia or myelodysplastic syndrome. FISH analysis showed that all 11q breakpoints were located centromeric to the MLL gene and between cosmids CN2900 and CN1323. We identified a yeast artificial chromosome (YAC) clone that spanned the inv(11) breakpoints on 11q. From this YAC, we identified a P1 clone, which included the breakpoints in at least three of the four patients. It is highly likely that the same gene on the P1 clone is rearranged in leukemic cells of each patient. This gene may be one of the targets for topoisomerase II inhibitors. PMID- 9218996 TI - Two discrete regions of deletion at 7q in uterine leiomyomas. AB - This study further defines the region of consistent deletion of chromosome 7 in uterine leiomyomas. We have examined 74 leiomyomas for allelic loss of markers spanning the 7q22 region defined by markers D7S518 and D7S471. Forty tumors with cytogenetically defined 7q deletions, twenty-nine tumors without cytogenetically visible 7q deletions, and five tumors with no cytogenetic information were examined for allelic loss of D7S518, D7S666, D7S515, D7S658, D7S496, D7S692, and D7S471. Loss of heterozygosity for one or more of these loci was observed in twenty-eight leiomyomas with cytogenetically defined 7q deletions and in three leiomyomas with a normal karyotype. Allelic loss of D7S666 was common and was observed in all twenty-three informative tumors with 7q deletions and in two tumors with normal karyotypes. This study indicates the presence of a tumor suppressor gene in close proximity to the D7S666 locus. Eight tumors followed an unusual pattern of allelic loss. These tumors showed retention of heterozygosity for at least one locus flanked by deleted loci. These results suggest the possibility that two discrete regions of deletion at 7q22 are involved in the development of a subset of leiomyomas. PMID- 9218997 TI - Frequent gain of copy number on the long arm of chromosome 20 in human pancreatic adenocarcinoma. AB - We have used comparative genomic hybridization (CGH) to survey genomic regions with aberrant copy numbers of DNA sequences in pancreatic adenocarcinoma. In 12 cell lines and 6 primary tumors from 18 patients with pancreatic adenocarcinomas, highly frequent losses (> 60%) were observed on chromosome arms 6q, 9p, and 18q and the Y chromosome. Moderately frequent losses (40-60%) were observed on chromosome arms 3p, 4q, 8p, and 21q. Interestingly, these samples showed extremely high frequencies of increases in copy numbers of DNA sequences on the long arm of chromosome 20 (15/18, 83%). We further analyzed five cell lines by fluorescence in situ hybridization (FISH) with probes on chromosome 20 to define the increase in copy number more accurately, and we found that 20q was increased to between 5 and 8 copies per cell. These results suggest the existence of an oncogene or oncogenes in 20q that play a role in the development and/or the progression of pancreatic carcinogenesis. PMID- 9218998 TI - Regenerative lesions in ulcerative colitis are characterized by microsatellite mutation. AB - An increased risk of colon cancer has been observed in individuals with long standing ulcerative colitis (UC). In order to identify molecular genetic markers for the development of neoplasia in UC individuals, we isolated DNA from normal, regenerative, and dysplastic mucosa, as well as from colon carcinomas from UC patients, and evaluated it for the presence of mutations in microsatellite DNA sequences. DNAs isolated from regenerative mucosa displayed microsatellite mutation. These observations suggest that DNA mutation is an early event in the UC disease process. PMID- 9218999 TI - Novel regions of chromosomal loss in familial neuroblastoma by comparative genomic hybridization. AB - Childhood neuroblastoma, an embryonal neoplasm of sympathetic nervous system progenitors, occurs in a familial form with an autosomal dominant mode of inheritance. Genetic susceptibility to this disorder is thought to arise via a germline mutation affecting a tumor suppressor gene, in accord with the two-hit model established for familial and sporadic retinoblastoma. Surprisingly, the familial neuroblastoma predisposition locus does not map to chromosome band 1p36, a genomic region likely to contain one or more neuroblastoma suppressor genes. We reasoned that inherited point mutations affecting one allele would be unmasked in many cases by somatically acquired deletions of the second allele that included the target gene in the tumor cells from these patients. Thus, to identify chromosomal regions that might contain suppressor genes important in hereditary neuroblastoma, we analyzed six familial tumors by comparative genomic hybridization. Recurrent losses of genetic material were detected on chromosome arms 3p (consensus region, 3p24-pter), 10p (consensus, 10p12-p13), 10q (consensus, 10q25-qter), 16q (consensus, 16q12-q22), and 20q (consensus, 20q13.3 qter), in addition to the regions commonly deleted in sporadic neuroblastomas (1p36 and 11q). These chromosomal sites may harbor novel tumor suppressor genes that could aid in our understanding of the predisposition to and pathogenesis of familial neuroblastoma and potentially sporadic tumors as well. PMID- 9219000 TI - Loss of heterozygosity on chromosome arm 16q in breast cancer metastases. AB - One of the main genetic abnormalities associated with breast carcinogenesis is the loss of genetic material from chromosome arm 16q. Different groups have identified two regions (16q22.1 and 16q24-ter) that are frequently deleted in primary tumors, suggesting the presence of tumor suppressor genes in these regions. Little is known about the late stages of tumor progression in this respect, and we, therefore, analyzed biopsy specimens of breast cancer metastases for deletions in these critical regions of 16q. We examined fine needle cytopunctures from 24 metastases, each with lymphocyte DNA, for allelic imbalance on 16q by means of polymerase chain reaction (PCR) with 15 highly polymorphic markers. All the metastatic samples showed deletion of at least one informative locus on 16q. The loss of heterozygosity (LOH) pattern often indicated the loss of a complete long arm of chromosome 16 (13 cases); nevertheless, in the remaining 11 samples, partial LOH patterns were observed. A small region of overlap (SR02) in 16q22.1 was frequently involved, whereas another (SR01) in 16q24-ter was affected in only two cases. A third region of LOH in 16q22.2-q23.2 was found in 6/11 samples. These results suggest that at least three different regions are involved in allelic imbalance on chromosome arm 16q in breast cancer. Loss of material from the third region could be a major event in the genesis of metastases. PMID- 9219001 TI - Monosomy 22 in a mixed germ cell-sex cord-stromal tumor of the ovary. AB - We report the cytogenetic findings in a case of mixed germ cell-sex cord-stromal tumor of the ovary in a 5-month-old girl. Monosomy 22 was observed as the sole karyotypic abnormality. This result was confirmed by comparative genomic hybridization, which revealed no additional chromosomal imbalances. This is the first observation of a chromosomal aberration in a mixed germ cell-sex cord stromal tumor of the ovary. Monosomy 22 has been previously observed in granulosa cell tumors of the ovary. This could suggest a common pathogenetic pathway for both types of tumors. PMID- 9219002 TI - Unusual clonal chromosomal evolution in a breast carcinoma and its lymph node metastasis in a patient with Down syndrome. AB - A cytogenetic study was performed on a primary breast carcinoma and its axillary lymph node metastasis from a 53-year-old patient with trisomy 21, a carrier of a constitutional der(21;21). A translocation t(X;21) and the loss of the other X chromosome were shared by all karyotypes from tumor cells. The primary tumor was hyperdiploid with several gains of whole chromosomes. In contrast, most cells from the metastasis shared several rearrangements and losses leading to a hypodiploid karyotype. No normal chromosome 17 was present; instead, an i(17)(q10) and a fragment, detected by chromosome painting and presumably corresponding to a rearranged 17p, were found. Immunostaining for p53 was strongly positive in the metastasis but not in the primary tumor, suggesting a mutation of the TP53 gene in the metastasis. Finally, a small cell population of the metastasis was hyperdiploid like the clone in the primary tumor, suggesting that the node was colonized twice, at an early stage and a later stage of the clonal evolution of the tumor. PMID- 9219003 TI - Patterns of meiotic variability of the (CAG)n repeat in the Huntington disease gene. AB - We have collected 76 parent-offspring (CAG)n values in 60 French Huntington's disease (HD) pedigrees. The analysis of intergenerational alterations in CAG repeat length shows that there is a correlation between repeat instability and parental repeat length. Paternally inherited cases are characterized by a preferential trend towards an increase in range of repeat sizes in offspring of HD patients. PMID- 9219004 TI - Hereditary multiple benign cystic epithelioma (multiple trichoepithelioma) with onset at early age. AB - A mother and a daughter affected with multiple trichoepithelioma were studied. The age of onset of the symptomatology in both was 7-years-old, the daughter being more severely affected than the mother at this age. This early age of onset is an exceptional observation which could be explained by maternal imprinting. PMID- 9219005 TI - The oculo-dento-digital syndrome: male-to-male transmission and variable expression in a family. AB - We report two siblings--a 5 1/2 year old female and her 4 1/2 year old brother, both presenting the classical clinical findings of oculo-dento-digital dysplasia (ODD). 1. Digital anomalies: bilateral complete cutaneous syndactyly of fingers IV-V (III-IV-V at the left hand of the boy) and camptodactyly IV. 2. Facial and ocular anomalies: microphtalamos-epicanthal folds, small midfacies, thin nose with hypoplastic alae nasi and small nares. 3. Dental anomalies with partial dental agenesis and enamel hypoplasia. Examination of the parents showed a bilateral cutaneous syndactyly IV-V in the father as the sole partial manifestation of ODD. The findings in the present family confirm the autosomal dominant inheritance of ODD with great variability in clinical expression. Moreover, the facial morphology (thin, hypoplastic nose) observed in several ODD patients suggests nosological overlap with the Hallerman-Streiff syndrome and could indicate that both syndromes are variable expressions of a contiguous gene deletion syndrome. PMID- 9219006 TI - Partial trisomy 15q: report of a patient and literature review. AB - We report a girl with severe developmental delay, scoliosis and mild dysmorphism. She was found to have a partial duplication of the long arm of chromosome 15. Precise cytogenetic diagnosis was possible after additional in situ hybridisation. A Karyotype of 46,XX,dup (15) (pter-->q26.3::q24-->qter) was concluded. We compare her data with the literature. No specific phenotype was found. PMID- 9219007 TI - Deafness and Mondini dysplasia in Kabuki (Niikawa-Kuroki) syndrome. Report of a case and review of the literature. AB - Report of a case and review of the literature: We report the case of a seven-year old female kabuki patient suffering from severe bilateral deafness related to Mondini dysplasia and ossicular anomalies. A review of the literature in English confirms that hearing loss is a major component of Kabuki Syndrome (KS) with a frequency at around 32%. However the possible mechanisms have not been fully described and hearing loss is often attributed to otitis media, but one reported case had severe ossicular malformations, two had sensorineural deafness and three others had mixed deafness. Our observation is the first reported case of Mondini dysplasia in KS. Awareness by physicians of this problem has a major practical consequence as diagnosis of Mondini dysplasia implies searching for and surgical prevention and treatment of perilymphatic fistula in order to prevent meningitis. PMID- 9219008 TI - Penetrance of familial hypertrophic cardiomyopathy. AB - Familial hypertrophic cardiomyopathy (FHC) is an autosomal dominant cardiac disease for which the penetrance remains a much-debated issue. Since the recent identification of the genes involved in the disease, the penetrance of FHC has not been reassessed in a large genotyped population. The aim of our study was therefore to evaluate it, according to age and sex, in ten families with previously identified mutations. Among 178 individuals we studied, 90 were genetically affected (9 different mutations in 3 genes). We found that penetrance, assessed by classical echocardiographic and electrocardiographic criteria, was (1) incomplete: 69%; (2) age-related: 55% between 10 and 29 years old, 75% between 30 and 49 y. and 95% over 50 y.; (3) greater in males than in females: 77% vs 58%, age-adjusted odds ratio: 3.98, CI 95%: 1.34 to 11,48; (4) similar for the genes analyzed. The consequences of these results for genetic counseling and linkage analyses are discussed. PMID- 9219009 TI - Brachydactyly and short stature in a mother and her daughter with a fragile site at 16q22. AB - A girl with short stature and brachydactyly had a fragile site at 16q22. Her mother had the same phenotype and the fragile site, too. These cases open the discussion of a possible association between a rare fragile site and an abnormal phenotype. Fragile sites are divided into two major groups, those which are rate and those which are common. The fragile site at 16q22 is a rare inductible fragile site. It is possible that the association between brachydactyly and the fragile site in a mother and her daughter may be due to chance. However rare fragile sites predispose to phenotypic abnormalities. The abnormal features in these two cases could be the result of the disruption of a gene involved in a skeletal development. PMID- 9219010 TI - Hyperacusis in Williams syndrome: a sample survey study. AB - Williams syndrome is a true multiple congenital anomalies mental retardation syndrome affecting the vascular, connective tissue and the central nervous system. Affected individuals have a distinctive neuropsychological profile characterized by extremely poor visuospatial skills but relatively preserved verbal skills. A very striking characteristic is the hyperacusis or over sensitivity to particular sounds. Klein et al. (4) found high rates (95%) of auditory over-sensitivity in a sample of Williams patients. The cause and mechanisms of auditory-over-sensitivity in Williams syndrome remain unclear. Some association has been suggested between hyperacusis and the occurrence of otitis media and also between hyperacusis and hyperactivity. The present study reports the results of an investigation into the occurrence of hyperacusis, otitis media and hyperactivity in a large group (N = 82) of Dutch speaking subjects with Williams syndrome from Belgium and The Netherlands. Prevalence and characteristics of hyperacusis and co-occurrence with otitis media and hyperactivity will be discussed and some management strategies are offered. PMID- 9219011 TI - Brachytelephalangy with mental retardation, peculiar face and short stature in two sibs. A new MCA/MR syndrome? AB - A new MCA/MR syndrome?: Two sibs are described who shares as clinical features microcephaly, short stature, deep set eyes, thick eyebrow, straight long nose with prominent bridge, septum extending below alae nasi, short philtrum, small mouth, high palate, prominent everted lower lip, and brachytelephalangy. To the best of our knowledge, this association has not been reported before. PMID- 9219012 TI - On the phenotypic overlap between "severe" oto-palato digital type II syndrome and Larsen syndrome. Variable manifestation of a single autosomal dominant gene. AB - We report two familial cases of oto-palato-digital (OPD) Type II syndrome, a father and his son. This family shows that OPDII syndrome is inherited as an autosomal dominant condition. The similarities between the OPDII syndrome and the Larsen syndrome are discussed. PMID- 9219013 TI - A collaborative approach to the diagnosis of a lethal short limb skeletal dysplasia. AB - We report a male infant with a lethal short limb Skeletal Dysplasia, born in a District Hospital in the South of Portugal. Local paediatricians investigated clinical and radiographical data. At the Perinatal Pathology Unit of the Egas Moniz hospital the diagnosis of Atelosteogenesis type II was proposed by the Clinical Geneticist and was supported by histopathological findings. Only a collaborative approach turned possible the diagnosis of this unusual entity. Atelosteogenesis type II and Diastrophic Dysplasia are closely related diseases, with similarities in phenotypic and histopathological presentation. Recently, these similarities were extended to molecular levels. The DNA analysis, in progress, will be able to establish a final diagnosis for this affected family. PMID- 9219014 TI - Atrial septal defect in Hallermann Streiff syndrome. PMID- 9219015 TI - Democracy and compulsory vaccination. PMID- 9219016 TI - More on Martin. PMID- 9219017 TI - Two of a kind--or none? PMID- 9219018 TI - Medicine and public health, ethics and human rights. AB - There is more to modern health than new scientific discoveries, the development of new technologies, or emerging or re-emerging diseases. World events and experiences, such as the AIDS epidemic and the humanitarian emergencies in Bosnia and Rwanda, have made this evident by creating new relationships among medicine, public health, ethics, and human rights. Each domain has seeped into the other, making allies of public health and human rights, pressing the need for an ethics of public health, and revealing the rights-related responsibilities of physicians and other health care workers. PMID- 9219019 TI - Enhancing cognition in the intellectually intact. AB - As science learns more about how the brain works, and fails to work, the possibility for developing "cognition enhancers" becomes more plausible. And the demand for drugs that can help us think faster, remember more, and focus more keenly has already been demonstrated by the market success of drugs like Ritalin, which tames the attention span, and Prozac, which ups the competitive edge. The new drug Aricept, which improves memory, most likely will join them. Whether such drugs are good for individuals, or for society, is an open question, one that demands far more public discussion. PMID- 9219020 TI - Mistrust, racism, and end-of-life treatment. PMID- 9219021 TI - Giving scientists their due. The Imanishi-Kari decision. PMID- 9219022 TI - Why does removing machines count as "passive" euthanasia? AB - The distinction between "passive" and "active" euthanasia, though problematic and highly criticized, retains a certain intuitive appeal. When a patient is allowed to die, nature appears simply to be taking its course. Yet when a patient is killed by, say, a lethal injection, humans appear to be causing his or her death. Guilt seems to follow naturally from the latter act while not from the former. Yet this view only holds up if age-old and vague ideas about "nature" and "artifice" go unscrutinized. Once examined more closely the functional relevance of particular machines to particular bodies becomes evident. And the innocence and guilt less clear. PMID- 9219023 TI - Deliberating about bioethics. AB - In some sense, bioethics was built on conflicts. Abortion, physician-assisted suicide, patients' demand for autonomy all are staple and contentious issues. And the controversies continue to proliferate. What forum best serves such debates? A look at political theories of democracy can help answer that question. The most promising for bioethics debates are theories that ask citizens and officials to justify any demands for collective action by giving reasons that can be accepted by those who are bound by the action. This conception has come to be known as deliberative democracy. PMID- 9219024 TI - Evaluating chemical risks: results of a survey of the British Toxicology Society. AB - 1. Members of the British Toxicology Society participated in a survey to determine their attitudes, beliefs, and perceptions regarding risks from chemicals. Similar surveys had previously been conducted with toxicologists and members of the general public in the United States and Canada. Data from 312 completed questionnaires were analyzed. 2. In general, the British toxicologists judged risks to be quite low for most hazards, with the exception of cigarette smoking and asbestos. They tended to have quite favorable attitudes toward the use of chemicals and were confident about the adequacy of chemical regulations. 3. As in previous studies of toxicologists, women expressed higher perceptions of risk than did men and had consistently stronger anti-chemical attitudes. 4. Toxicologists working in industry had more favorable attitudes towards chemicals and their use than did those working in academic settings. 5. When asked to evaluate chemical technical summaries of various animals studies there was considerable disagreement among the respondents about the toxicity of the chemicals involved. 6. In general, British toxicologists were equivocal about the reliability of animal studies in predicting human effects (particularly carcinogenicity) probably because of the belief that animal studies overestimate risk. However, they were rather confident that human health risks could be assessed reasonably accurately. PMID- 9219025 TI - Effects of oxygen pressure and medium volume on the toxicity of paraquat in rat and human type II pneumocytes. AB - The herbicide, paraquat is highly toxic for mammals, with the lungs being the main target organ, because of the active accumulation of the compound in this organ. The cellular toxicity of paraquat has been shown to be an O2-driven process and hyperoxia is known to increase the lethality of paraquat. In this study we have examined the effect of various O2 concentrations on the toxicity of paraquat in rat and human type II pneumocytes in culture, and we have tested whether the thickness of the liquid layer above the cells would influence the toxicity of paraquat. Type II pneumocytes were isolated from rat or human lung tissue using trypsin digestion, percoll density gradient centrifugation and differential attachment. Adherent cells (day 2) were incubated for 20 h in different volumes of culture medium (thickness of liquid layer), whether or not in the presence of paraquat, in the presence of different O2 tensions. The viability of the cells was assessed by the release of LDH in the culture medium. In both rat and human type II pneumocytes the toxicity of paraquat was independent of the thickness of the liquid layer (2.5 to 10 mm height). The toxicity of paraquat in rat type II pneumocytes decreased from a TC50 value of 28 microM paraquat at 21% O2 to 107 microM at 10% O2 and increased to 12 microM and 8 microM at 60% and 85% O2, respectively. For human type II pneumocytes the TC50 values were 7 microM; 25 microM and > 1000 microM paraquat at 60%, 21% and 10% O2, respectively. In this study we have shown that the diffusion of O2 through a liquid layer does not limit the toxicity of paraquat and that, as in vivo, increasing O2 partial pressure enhances the toxicity of paraquat. PMID- 9219026 TI - Effects of high-level exposure to lead on NK cell activity and T-lymphocyte functions in workers. AB - 1. NK and T cell functions were measured in peripheral blood lymphocytes of workers occupationally exposed to lead. 2. No differences were found in these functions even in those workers with higher levels of blood and urine lead and urinary delta-ALA than the currently accepted biological limit values as compared to controls. 3. We conclude that high chronic exposure to lead is not associated with an impairment or either T- or NK cell functions in man. PMID- 9219027 TI - Study of oxidative-stress in rifampicin-induced hepatic injury in growing rats with and without protein-energy malnutrition. AB - 1. Rifampicin (RMP) induced hepatic injury was investigated in growing rats. The interaction of moderate and severe protein-energy malnutrition (PEM) was also investigated. 2. Status of oxidative/antioxidative profile was studied by the mechanistic approach, to enumerate the nature of injury. 3. Successful hepatic injury in rats was produced by giving intraperitoneal injection of RMP (50 mg/kg/day). 4. Hepatic lipid peroxidation was significantly increased in all the RMP treated rats. 5. Superoxide dismutase, catalase and glutathione peroxidase activities in the hepatic tissue decreased with RMP treatment. 6. Hepatic thiols represented as total and protein-bound thiols, showed significant elevation, whereas the non protein thiols remain unchanged with RMP treatment. 7. Glutathione-S-transferases also showed significant elevation against 1,2-dichloro 4 nitrobenzene (DCNB) and ethacrynic acid (EA) as substrates. 8. The oxidative/antioxidative profile was observed to be more severely affected with coexistence of malnutrition. 9. Histopathological correlation showed an additional fatty infiltration of hepatocytes with coexistence of malnutrition. 10. Thus, in conclusion, it can be speculated that an altered oxidative/antioxidative profile is the closely associated with production of RMP induced hepatic injury. PMID- 9219029 TI - Changes in the arterial blood pressure, heart rate and normal ECG parameters of rat after envenomation with Egyptian cobra (Naja haje) venom. AB - 1. The effect of Egyptian cobra (Naja haje) venom on the normal electrical activity of the cardiac muscles (ECG) and arterial blood pressure of envenomated rats were investigated in this study. 2. Rats were divided into three groups. The first group was injected im with saline and considered as control group. Rats of the second and third groups were injected IM with 0.02 micrograms and 0.04 micrograms cobra venom/gim b.wt, respectively. 3. Mean blood pressure (MBP), heart rate (HR) and four different ECG parameters (PR and QT intervals, R and T wave amplitudes) were measured over 1 h following envenomation. 4. The low dose (0.02 micrograms/g) of N. haje venom caused hypotension accompanied by an increase in the HR, whereas hypertension and bradycardia developed after injection of the high dose (0.04 micrograms/g) of venom. 5. There was a decrease in the P-R interval after administration of the low dose and prolongation of it after the high dose. The Q-T interval and R-wave amplitude were significantly increased after injection of both doses. T-wave amplitude was significantly elevated only after injection of the high dose. 6. The present results indicate that the Egyptian cobra (N. haje) venom significantly alters the arterial blood pressure and ECG parameters of envenomated rats. The suggests that impairment of the electrical activity of cardiac muscle may be one of the reasons why victims of cobra bite die. PMID- 9219028 TI - Defective neutrophil function in workers occupationally exposed to hexachlorobenzene. AB - In this work we have studied the respiratory burst and chemotaxis of polymorphonuclear leukocytes from 51 workers exposed to chlorinated compounds, which were compared with those of non-exposed, age- and sex-matched individuals. These two neutrophil functions were significantly reduced as compared to controls. No correlation was observed between the length of exposure, hexachlorobenzene (HCB) blood concentrations and neutrophil chemotaxis or the extent of nitroblue tetrazolium reduction. PMID- 9219030 TI - Trichloroethylene radicals generated by ionizing radiation. An EPR/spin trapping study. AB - Trichloroethylene (TCE) was exposed in the presence of the spin trap N-tert-butyl alpha-phenyl nitrone (PBN, 0.1 M) to ionizing radiation from two different sources in an attempt to determine the origin of the spin-trapped radicals generating the EPR spectra in precision cut liver slices. TCE samples were irradiated with 18 MeV electrons to a total dose of 1000 Gy in a linear accelerator (LINAC) or exposed to 60Co gamma-rays to total doses of 100 Gy and 1000 Gy. The results show that three PBN adducts were generated during the LINAC radiations. Two of these spin adducts correspond to the addition of carbon centered radicals to PBN, and the third adduct is consistent with a decomposition product of PBN. The predominant carbon-entered radical yields a PBN adduct that is more stable, persists for over 24 h and has identical hyperfine coupling constants (aN = 1.61 mT, aH beta = 0.325 mT) to the PBN adduct obtained when precision-cut liver slices were exposed to TCE. Gamma radiation (100 Gy) of TCE yields PBN adducts with lower primary nitrogen hyperfine coupling constants (aN = 1.45 mT and aN = 1.54 mT). The results (gamma-radiation) suggest that the carbon centered radical is formed on a single TCE carbon that is different than the predominant radical formed during LINAC radiations. This difference is confirmed by experiments using 13C-TCE. The results further suggest that, during gamma radiation of TCE, the radicals are formed by dechlorination at the TCE carbon containing two chlorine atoms. The results obtained during LINAC radiations suggest that the predominant radical is formed by dechlorination at the TCE carbon containing a single chlorine and a single proton. In addition, it is possible that this radical is the initial TCE radical formed during exposure of liver slices to TCE. PMID- 9219031 TI - Modulation of soluble CD40 ligand bioactivity with anti-CD40 antibodies. AB - The B cell surface molecule CD40 may be activated either by its ligand CD40L or by anti-CD40 antibodies. In this study, five new anti-CD40 monoclonal antibodies (MAb) were characterized. Bioactivity of the MAb was assessed using a receptor hybrid consisting of the extracellular domain of CD40 and the intracellular domain of the p55 TNF receptor as a model for CD40 activation. Two agonistic MAb were able to enhance the activation of this CD40 hybrid CD40L. These MAb bound to an epitope that was not located within the CD40L-binding region indicating that activation of CD40 occurs epitope-independent. A second pair of ligand mimetic anti-CD40 MAb which appeared to bind to the CD40L binding site decreased CD40L bioactivity. With regard to ligand mimetic effects binding of the CD40L epitope was not of advantage. Combining anti-CD40 MAb with different epitope specificities or cross linking anti-CD40 MAB with secondary antibodies enhanced ligand mimetic effects. These data clearly show that ligand or antibody-mediated receptor aggregation is the major mechanism by which CD40 is activated. Furthermore, our data support that an aggregate of activated receptors is favorable in regard to CD40 activation. PMID- 9219032 TI - Primary structure and functional scFv antibody expression of an antibody against the human protooncogen c-myc. AB - The immunoglobulin heavy- and light-chain variable region (Vh and Vl) genes were isolated from Myc1-9E10 hybridoma cells, which secreted monoclonal antibody against human oncogen c-myc. The expression vector pOPE52-c-myc was constructed for the recombinant production in E. coli. A 30 kDa single chain fragment (scFv) expression product was found in the periplasmic space by SDS-PAGE and immunoblotting. A significant fraction was processed correctly as demonstrated with an antiserum recognizing the processed aminoterminus only. The specific binding of the scFv fragment to the peptide epitope of the maternal monoclonal antibody was demonstrated and the primary sequence of the variable regions was determined. Sequence comparison with previously published partial Vh and Vl sequences from this hybridoma cell line revealed a genetic heterogeneity for the light chain variable region. The potential use of this scFv as a new tool for detection and purification of tagged proteins, for adding costimulatory signals to the surface of cancer cells as well as for analyzing c-myc function in the living cell by cytoplasmic expression is discussed. PMID- 9219033 TI - Primary structure and functional expression of heavy- and light-chain variable region genes of a monoclonal antibody specific for human fibrin. AB - The immunoglobulin heavy- and light-chain variable region (VH and VK) genes were isolated from 8E5 hybridoma cells, which secreted monoclonal antibody against human fibrin by RT-PCR. An expression vector pOPE51-8E5 was constructed for the recombinant VH-VK scFv expression. The primary sequence of the variable regions was determined. Expression product was found in the periplasmic space and inclusion bodies by SDS-PAGE and immunoblotting. It was a 30 KDa single chain fragment (scFv) with the antigen-binding specificity of the parental monoclonal antibody. A light chain shuffling with an unspecific VL did not result in a loss of fibrin binding specificity. PMID- 9219034 TI - Biodistribution of anti-breast mucin HuBrE3-derived inverted Fabs (IFabs) in nude mice carrying MX-1 xenografts. AB - In radioimmunotherapy, the long circulation times of antibody radioconjugates correlate with high relative radiation doses to nontumor tissues. Tumor/normal tissue ratios can be significantly improved by using targeting molecules with shorter circulation times. IFabs are multimers of VH-CH1-linker-VK-CK monomers. The lack of the Fc region in IFabs should lead to circulation times that are shorter than those of IgG molecules. The monomers assemble into disulfide-bond stabilized multimers, 90% of which are 100 kDa dimers (IFab2). IFab2s should not be rapidly eliminated through kidney filtration because their molecular weight is above the threshold for renal passage. We report the first experimental in vivo tests for 125I-IFab radioconjugates derived from a humanized version of the anti breast mucin monoclonal antibody BrE-3. Biodistributions are reported for athymic nude mice carrying human mammary tumor MX-1 xenografts. The T1/2 beta's for the different tissues ranged from 13.3 h for blood to 19.9 h for tumor. Therefore, IFab radioconjugates cleared the body with a rate comparable to that of F(ab')2 fragments. Except for stomach, tumor/nontumor dose ratios were significantly better for IFabs than for the parent antibody (BrE-3)4 days after injection. PMID- 9219035 TI - Characterization of a novel adhesion function blocking monoclonal antibody to rat/mouse P-selectin generated in the P-selectin-deficient mouse. AB - P-selectin is an important adhesion molecule involved in leukocyte migration. However, to date, no monoclonal antibodies (MAb) generated against rat P-selectin have been identified which block P-selectin mediated leukocyte adhesion. Most studies in the rat have utilized crossreacting antibodies generated against P selectin in higher species. In a P-selectin deficient mouse we generated an anti rat/mouse P-selectin MAb, designated RMP-1, by immunization with activated rat platelets. This IgG2a MAb immunoprecipitates a 140 kDa protein under reducing conditions from rat platelet lysate. By ELISA and immunofluorescence flow cytometry, MAb RMP-1 reacts with thrombin-activated but not unactivated rat platelets. In addition, by ELISA MAb RMP-1 binds to activated mouse platelets and recombinant rat and mouse P-selectin. MAb RMP-1 inhibited adhesion of HL-60 myeloid cells to immobilized mouse P-selectin by 97% and to activated rat and mouse platelets by 100% under static conditions, confirming the adhesion function blocking activity of MAb RMP-1. This novel MAb should be useful for studying P selectin function in vitro and in vivo in both rat and mouse inflammation models. PMID- 9219036 TI - Monoclonal antibodies specific for peptide epitopes of the epidermal growth factor receptor's extracellular domain. AB - The ErbB tyrosine kinase receptor family plays an important role in normal cellular growth and differentiation. In addition, ErbB receptor family members are commonly amplified and overexpressed in various human neoplasms and tumor derived cell lines, where it is believed that increased signalling as a result of receptor overexpression may play an important role in oncogenesis. Consequently, ErbB receptor family members are being investigated rigorously as potential biomarkers of cancer and as therapeutic targets in malignant tissues. Numerous studies now demonstrate the existence of "soluble" ErbB (sErbB) analogs in normal and cancerous tissues. These sErbB proteins embody the extracellular domain (ECD) of the receptor only; they are generated by either proteolytic cleavage or from truncated, alternatively spliced mRNA transcripts. Recently, we have identified an alternate transcript of the human c-erbB1 (Epidermal Growth Factor Receptor) proto-oncogene from placenta that encodes a sErbB1 protein of 60-kDa. This protein, p60 sErbB1, is glycosylated and secreted when expressed in transfected tissue culture cells in vitro. Although "soluble" receptor analogs may play important physiological roles in intercellular communication, tissue morphogenesis, tissue regeneration and repair, and embryogenesis by inhibiting or stimulating specific mitogenic and pattern forming signals, their mechanism of action has not been thoroughly elucidated. To further characterize sErbB1 expression in human tissues and cell lines and to better understand their role in carcinogenesis and normal development, we have generated monoclonal antibodies (MAbs) toward specific peptide epitopes of ErbB1 extracellular subdomains III and IV. These antibody reagents are described here and should be useful experimental, preparative, analytical, diagnostic, and therapeutic reagents for the study of sErbB1 molecules in normal development and cancer. PMID- 9219037 TI - Monoclonal antibody NM11 recognizes a C-terminal epitope shared by p300 and CBP. AB - The epitope recognized by the monoclonal antibody NM11, previously shown to recognize both CBP and p300, has been mapped here to the C-terminal third of p300 and CBP by Western analysis of p300 and CBP prokaryotic fusion proteins. More precise epitope mapping, carried out by screening a plasmid expression library derived from small randomly generated CBP cDNA fragments localizes the NM11 epitope to a 21 amino acid stretch spanning amino acids 2071-2091 near the CBP C terminus. CBP and p300 differ by three noncontiguous residues within this 21 amino acid region, a difference that does not detectably affect the reactivity of NM11. PMID- 9219038 TI - Monoclonal anti-fosB antibody specific for predetermined, nonstructural region of the fosB protein. AB - Comparison of the primary structures and theoretical prediction of the potential antigenic determinant of the deduced Fos proteins reveals the presence of a nonstructural and hydrophilic region juxtaposed to the leucine zipper and nonconserved among the Fos protein family. To develop monoclonal anti-peptide antibodies capable of distinguishing all Fos-proteins, synthetic peptides specific for the mentioned predicted region were synthesized manually by the "tea bag" method. Immunization of Balb/c mice with fosB-related synthetic peptide BSA gave rise to mouse hybridoma cell line K21 (IgG1, kappa) secreting highly specific antibodies against corresponding human fosB protein. Fine mapping of the MAb K21 indicated that the minimal epitope essential for the recognition is the sequence GPGPLAE. PMID- 9219039 TI - Functional characterization of monoclonal antibody inhibitors of alpha 2 antiplasmin that accelerate fibrinolysis in different animal plasmas. AB - In humans with acute thrombotic disease, thrombi often appear to resist fibrinolysis induced by plasminogen activators. To examine the potential role of alpha 2-antiplasmin (alpha 2AP) in thrombus resistance in vivo, we generated monoclonal antibody inhibitors of alpha 2AP. In a somatic cell fusion, 99 hybridomas were obtained that produced MAbs that bound to human 125I-alpha 2AP in a capture assay. Screening assays showed that 3 of these MAbs, 49, 70, and 77, neutralized the function of alpha 2AP. Immunoblotting experiments indicated that these MAbs recognized an epitope present in native alpha 2AP that was destroyed by denaturation with SDS. Each of these MAbs fully inhibited the binding of the other MAbs to alpha 2AP, but none of them competed with the binding of another anti-alpha 2AP MAb, RWR. When tested for their binding to nonhuman alpha 2APs in plasmas, all three MAbs were strongly crossreactive with all primate plasmas tested but showed an idiosyncratic pattern of binding to alpha 2AP in other plasmas, suggesting unique fine epitope specificities. In human plasma, all three MAbs amplified the lysis of human plasma clots induced by plasminogen activators, increasing the potency of urokinase by nearly 50- to 100-fold. These MAbs also markedly amplified the lysis of clots from baboon, cynomolgus, african green monkey plasmas, and to a lesser extent, ferret and dog. By virtue of their ability to potently inhibit alpha 2AP in other animal plasmas, these MAbs should be useful for examining the role of alpha 2AP in thrombus resistance to fibrinolysis in vivo. PMID- 9219040 TI - Rapid detection of bovine type II collagen-specific T-cell hybridomas. AB - T-cell hybridomas are powerful tools in studying the fine specificities of antigen recognition by the T-cell receptor (TCR), the structure and genetic basis of the CD3-TCR complex, and the size of the TCR alpha/beta repertoire used in response to various antigens. A technical challenge in establishing T-cell hybridomas is the early identification of antigen-specific ones. We have established a rapid and efficient ELISA method for detecting antigen-specific T cell hybridomas. Our ELISA technique significantly reduces the time and resources required for the primary screening of antigen-specific T-cell hybrids, eliminates the need of maintaining hundreds of rapidly growing nonspecific clones, and does not require the maintenance of IL-2/IL-4 dependent cell lines such as CTLL-2 or HT-2. In addition, the ELISA technique is designed to detect both types of CD4 T cells: Th1 and Th2, by using a mixture of anti-IL-2 and anti-IL-4 monoclonal antibodies. Therefore, we believe that our ELISA technique provides a faster, less expensive, and higher throughput screening method for the early identification of antigen-specific T-cell hybridomas than the current bioassays. PMID- 9219041 TI - The role of antipsychotic agents in the treatment of bipolar disorder patients. AB - Antipsychotic agents have been an important adjunct in the acute and at times prophylactic management of bipolar disorder patients. As a result of safety considerations, particularly the risk of tardive dyskinesia, there are concerns about their long-term use as adjunctive treatment. Although alternative agents have been proposed, some treatment-refractory cases may not respond as well to other regimens. This sub-group of patients has not been well characterized, but may result from overinclusive criteria in defining bipolar disorder. A comprehensive Medline search dating back to 1966 was conducted to identify published studies on antipsychotic utilization in bipolar disorder patients. We investigated the support for their use, and their effectiveness in this population, and attempted to define the group of patients requiring antipsychotic treatment. Additionally, we reviewed specific safety considerations, and suggested standards for their use in bipolar patients. PMID- 9219042 TI - Long-term treatment of depression: is there a use for depot antidepressants? AB - Depot neuroleptics have been widely introduced for long-term treatment of schizophrenia. The question of whether depot antidepressants should be developed for the treatment of chronic depression and for the prophylaxis of recurrent depression is addressed. This approach seems to be indicated in patients showing poor compliance to oral antidepressive medication and in patients suffering from secondary depression and who are already receiving depot antipsychotics, but it is also indicated in subgroups of patients who, for social, cultural or personality reasons, have problems with regard to a regular and long-term intake of oral medication. Before the development of depot antidepressants is initiated, the ethical issues relating to this form of medication should be discussed. Technically, the preparation of depot forms may represent a serious challenge. Depot antipsychotics are all esters of hydroxylated neuroleptics and long-chain fatty acids. Potential candidates for antidepressants are venlafaxine, flesinoxan and E-10-OH-nortriptyline. The possibility of developing forms for transdermal application is also discussed. In conclusion, clinical considerations support the idea of the usefulness of a depot preparation for antidepressants but ethical and technical aspects should not be neglected. PMID- 9219043 TI - Paroxetine and pindolol: a randomized trial of serotonergic autoreceptor blockade in the reduction of antidepressant latency. AB - A double-blind, randomized, placebo-controlled, parallel group study was performed in 80 adult outpatients meeting ICD-10 criteria for major depression and with a Montgomery-Asberg Depression Rating Scale (MADRS) score of at least 18 at baseline. All patients received paroxetine (20 mg once a day) plus either pindolol (2.5 mg three times a day) or matching placebo for 6 weeks. Analysis of the day 14 MADRS scores on an intent-to-treat basis revealed a treatment-by centre interaction, with a significant effect of pindolol being demonstrable at only one centre. At this centre, 25% of the paroxetine plus pindolol group and 0% of the paroxetine plus placebo group showed a decrease of at least 50% from baseline MADRS by day 4 (p < 0.05). At day 14, the proportions were 73% and 7%, respectively (p < 0.001). Analysis of covariance on a "perprotocol" population demonstrated a significant accelerator effect of pindolol at days 4 and 7 in the absence of a treatment-by-centre interaction, but a centre effect was apparent at later time-points. The results suggest that the latency of antidepressant action can be reduced with pindolol augmentation. A large multicentre study is in progress to investigate this effect further. PMID- 9219044 TI - Fast onset: an open study of the treatment of major depressive disorder with nefazodone and pindolol combination therapy. AB - Pindolol, a beta-adrenergic and presynaptic 5-HT1 vA antagonist, when added to specific serotonin reuptake inhibitors, potentiates the antidepressant action, leading to an earlier onset of effect. Following on from the suggestion that nefazodone, a specific serotonin reuptake inhibitor and antagonist of 5-HT2, improves 5-HT1A-mediated transmission, we used a pindolol and nefazodone combination treatment for major depressive disorder. Twenty outpatients underwent a 4-week trial. Patients were seen twice a week, and completed efficacy and safety measures including the 17-item Hamilton Depression Scale, the Montgomery Asberg Depression Rating Scale and the Clinical Global Impression scales. Results demonstrated significant improvement in all efficacy measures after one visit (2 4 days of treatment), with decreasing depression scores on all measures continuing throughout the trial. After 1 week of treatment, 15 out of 20 patients had experienced a 50% or greater reduction in their 17-item Hamilton Depression Scale scores. Remission rates were dramatic, with 40% of patients in remission after 1 week of treatment and 90% after 4 weeks. This open study of nefazodone pindolol combination therapy suggests that this may be a new treatment option for patients with major depressive disorder; however, it needs to be replicated in a double-blind trial before conclusions regarding efficacy and safety can be made. PMID- 9219045 TI - Milnacipran, a new serotonin and noradrenaline reuptake inhibitor: an overview of its antidepressant activity and clinical tolerability. AB - Milnacipran (Ixel) is a new antidepressant with essentially equal potency for inhibiting the reuptake of both serotonin and noradrenaline, with no affinity for any neurotransmitter receptor studied. A review of the studies comparing milnacipran, placebo and active comparator antidepressants provides clear-cut evidence of its efficacy in both severe and moderate depression in hospitalized and community settings. Meta-analyses of the original data of controlled trials involving 1032 patients, comparing milnacipran with imipramine or selective serotonin reuptake inhibitors (SSRIs), show that milnacipran provides antidepressant efficacy similar to that of imipramine and significantly superior to that of the SSRIs. An analysis of a database of over 3300 patients shows that both the general and cardiovascular tolerability of milnacipran are superior to those of the tricyclic antidepressants (TCAs) with fewer cholinergic side effects. The tolerability of milnacipran was comparable to that of the SSRIs, with a higher incidence of dysuria with milnacipran, and a higher frequency of nausea and anxiety with the SSRIs. Milnacipran is a new therapeutic option in depression, which offers a clinical efficacy in the range of the TCAs combined with a tolerability equivalent to that of the SSRIs. PMID- 9219046 TI - Response to clozapine in acute mania is more rapid than that of chlorpromazine. AB - The purpose of the present study was to compare the efficacy of clozapine with that of chlorpromazine in an open label manner (both given in association with lithium salts) in the treatment of acute mania. Thirty hospitalized manic patients were entered into the study. All patients met DSM-IV criteria for bipolar disorder, Manic Episode; 27 patients completed the study and three patients dropped for noncompliance. The duration of the study was 3 weeks. Patients were randomly assigned to two treatment groups; group 1 (n = 15) was treated with clozapine at a mean dose of 166 mg/day and group 2 (n = 12) was treated with chlorpromazine at a mean dose of 310 mg/day. Manic symptomatology was rated on Young Rating Scale for Mania (YRSM) each week; side effects were recorded on dosage records and treatment emergent symptoms; extrapyramidal acute side effects were rated on the Simpson-Angus Rating Scale performed at the beginning of the study and after 3 weeks of treatment. A two-way repeated measures analysis of variance on YRMS scores showed a significant time effect (p < 0.0001) and a significant time-group interaction (p < 0.0001). Post-hoc comparison between the two groups showed a significant difference after 2 weeks of treatment (p = 0.0001), with clozapine treated patients showing lower YRSM scores than chlorpromazine treated patients. YRSM scores at the end of the study were not significantly different. Patients treated with clozapine showed a more rapid trend toward amelioration. No clinically relevant side effect was observed during the study. PMID- 9219047 TI - Diltiazem, a calcium antagonist, partly attenuates the effects of dextroamphetamine in healthy volunteers. AB - Calcium antagonists have previously been shown to be effective in the treatment of mania. In the present study we used dextroamphetamine administered to humans as a model of mania, to determine whether the calcium antagonist diltiazem would prevent dextroamphetamine-induced changes. This may help determine whether diltiazem is likely to be useful in the treatment of mania. Ten healthy volunteers were enrolled in this double-blind, placebo-controlled, balanced crossover study. Subjects received either oral diltiazem (60 mg) and placebo, placebo and dextroamphetamine (20 mg), diltiazem and dextroamphetamine, or placebo alone. Subjective and sleep changes were measured using visual analogue scales. Attentiveness and visual reaction times were measured repeatedly as were diastolic and systolic blood pressure. The results showed that dextroamphetamine alone produced a number of subjective changes, cardiovascular changes and changes in reaction time. Diltiazem significantly attenuated the cardiovascular changes, but not the subjective or reaction time changes. It is hypothesized that these findings may represent effects of diltiazem on noradrenergic neurotransmission. The results are tentatively supportive of suggestions that diltiazem may be clinically useful in the treatment of mania, as is another calcium channel antagonist, verapamil. PMID- 9219048 TI - Citalopram-associated clitoral priapism: a case series. AB - Priapism is documented to occur in association with treatment with a variety of psychotropic agents. Three cases of clitoral priapism associated with treatment with citalopram are presented. PMID- 9219049 TI - Listening in the care environment--chaos or clarity for the hearing-impaired elderly person. AB - The reported study evaluates listening circumstances within long-stay wards in an attempt to understand the nature of the environment and so develop and subsequently evaluate a programme of nursing intervention for hearing-impaired residents. Evidence is presented which indicates that improvement of the residents' listening environment, promotes adaptation to hearing aids and enhances residents' quality of life. It is recommended that attending to the listening environment becomes a concern of nurses working within continuing care facilities for elderly people. PMID- 9219050 TI - Perceptions of the body interior by children with asthma and children with no known chronic disease. AB - Comparatively little is known about how children perceive their internal bodies. In particular there is a lack of British studies, both of children's normal perceptions and in children experiencing chronic ill health. Such studies are important if information is to be given to children in the most effective way. The current study presents the results of an investigation of 72 children, 36 of whom were chronic asthma sufferers, whilst the others had no known chronic disease. Children at ages 5-6 years, 7-8 years and 10-11 years were asked to draw their internal body parts and to answer questions about the functioning of nine body organs. Data were analysed to examine the effect of age, sex and presence of asthma on the children's responses. Five-year-olds knew significantly less about their lungs than did other groups, boys identified muscle and ribs more frequently than girls, and there was an increase in levels of conceptualisation with age. There were no significant differences between asthmatic and non asthmatic children. The findings are considered with particular reference to their importance for nursing practice. PMID- 9219051 TI - Dimensions of care in five United States nursing homes: identifying invisible work in care-giving. AB - This article reports on a study carried out to identify components of practical knowledge, which is the knowledge that is used to guide decision-making and actions, used by care-givers in nursing homes. The practical knowledge behind the giving of care in the day-to-day activities in five nursing homes was studied and models for such care were developed. One finding from this study was that a large portion of the work of caring as known and understood by care-participants (patients, families, and nursing staff) was excluded from prescribed routines established by administration for care. PMID- 9219052 TI - Occupational prestige for registered nurses in the Asia-Pacific region: status consensus. AB - Occupational prestige for nurses may result in improved job satisfaction, enhanced abilities in health promotion activities, and autonomy in decision making related to patient care. Regional consensus in the ascription of professional occupational status and consequent prestige, between health workers and within registered nurses, has been investigated within the Asia-Pacific region and other countries. Consensus for registered nurses and medical officers at the level of professional was supported for 66% of countries. Australia, although rapid and complete in its transition to degree level education, still reflects a paraprofessional status compared with the professional status of medical officers. Registered nurses from Thailand and the Philippines led the region in within-group consensus with 80% of registered nurses holding a degree as their minimum educational qualification. Findings highlight the need to promote professional status for all registered nurses within the Asia-Pacific region and outline areas for potential disjuncture between status and prestige achievement. PMID- 9219053 TI - Constructing international professional identity: what psychiatric nurses talk about on the Internet. AB - The Internet is in the very early stages of being exploited by psychiatric nurses. Following an explanation of e-mail lists, this paper describes an analysis of the first 16 months operation of such a list devoted to psychiatric nursing. Those topics having international resonance for psychiatric nurses were: nursing models, labelling theory, the relationship of psychiatric nursing to medical psychiatry, the risks of depersonalised care, community care, and service cutbacks. Costs and benefits of the e-mail list to individual psychiatric nurses are described and evaluated. It is concluded that e-mail lists have the capacity to contribute to an international psychiatric-nursing identity in a unique and highly personal way. The Internet has many more possibilities for exploitation by psychiatric nurses. PMID- 9219054 TI - 'Maintaining the balance'--nursing care of patients with chronic heart failure. AB - Nurses in different settings are involved in caring for patients with heart failure. In the clinic, hospital, nursing home or patients' home, the nurse has a role in detecting, identifying and treating heart failure. In order to provide optimal care, literature on possible and effective interventions should be available to nurses. This study gives an overview of nursing care for patients with heart failure as described in practice, literature and standard nursing care plans. Based on interviews of 45 nurses, a review of all pertinent literature published between 1983 and 1993 and a review of standard nursing care plans used in the Netherlands to care for patients with heart failure, four composite themes emerged; namely, basic nursing care, assessment and observation, symptom relieving interventions, and patient education. Caring for patients with heart failure is very complex and is often aimed at keeping a very delicate balance between, e.g. rest and activity, fluid intake and elimination, and therapeutic cost and benefit. Treatment strategies for optimizing care for these patients are described. PMID- 9219055 TI - Measuring clinical nurse performance: development of the King's Nurse Performance Scale. AB - The development of the King's Nurse Performance Scale to measure clinical nurse performance is described. Instrument construction was informed by the Slater Nursing Competencies Rating Scale [Wandelt, M. A. and Stewart, D. S. (1975) Slater Nursing Competencies Rating Scale. Appleton-Century Crofts, New York] together with key literature and the use of expert opinion. The instrument was utilised to observe the clinical performance of senior student nurses (n = 99) and data which were at the ordinal level were statistically analysed using a variety of non-parametric tests. Key findings of students' observed nursing practice are presented in a separate paper (While et al., unpublished document). Internal consistency testing of the King's Nurse Performance Scale using Cronbach's alpha coefficient revealed a promising alpha for the total instrument (r = 0.93). The subsection alphas indicated that further refinement may enhance the strength of the instrument as a tool for the measurement of performance in different domains of practice. The possible use of the Scale in the professional development of newly qualified nurses is suggested. PMID- 9219056 TI - Measuring the quality of life of older persons: a model with implications for community and public health nursing. AB - Measuring the quality of life (QOL) of older persons can assist health professionals in achieving a number of important objectives. These include assessing the effects of illness and treatment, identifying need for support services, and developing health enhancing environments. Most QOL models focus unduly on illness and disability, define QOL too narrowly, and do not consider aspects of personal control and potential opportunities for change. A new model of QOL with associated instrumentation, the Quality of Life Profile: Seniors Version (QOLPSV), is described. Administration of the QOLPSV to 205 older persons in Ontario, Canada found it to be reliable and valid. Limitations of the instrument are presented and potential uses explored. PMID- 9219057 TI - Comparison of three methods for 17 alpha-hydroxyprogesterone. AB - We compared the performance of three 17 alpha-hydroxyprogesterone kits: the double antibody method, the coated tube method (ACTIVE), both from Diagnostic Systems Laboratories, Inc. (DSL) and the coated tube method (COAT-A-COUNT) from Diagnostic Products Corporation (DPC). The assay performance of the two DSL kits was very similar in terms of sensitivity, intra- and inter-assay precision, linearity of dilution, recovery, and specificity. We also analyzed 190 samples for 17 alpha-hydroxyprogesterone values using the above three kits. Twenty-three subjects were from prepubertal population (ages 1 month-13 years), thirty subjects were normal adult males (ages 20-53 years) and the remaining subjects were females in different phase of menstrual cycle (n = 40), on oral contraceptives (n = 20), post-menopausal (n = 17), or pregnant women in their first, second, or third trimester (n = 60). In addition to these 60 pregnancy samples, we analyzed serial samples from 3 pregnancies. 17 alpha-OHP levels paralleled the progesterone levels in all three kits. Although there was reasonable correlation between the DPC and the two DSL kits, the 17 alpha-OHP values were found to be significantly higher with DPC kit during the 2nd and 3rd trimester of pregnancy indicating probable interference in the DPC assay by some structurally related steroids present during pregnancy. The DSL assays may be particularly well suited for measuring 17 alpha-OHP levels during pregnancy. PMID- 9219058 TI - Association between expression of intercellular adhesion molecule-1 and integration of human T-cell-leukemia virus type 1 in adult T-cell leukemia cells. AB - It is known that the expression levels of intercellular adhesion molecule-1 (ICAM 1) in adult T cell leukemia(ATL) cells are high, whereas those in T-lymphoid cells are not. In order to investigate the factors that influence the induction of ICAM-1 molecules, Northern blot analysis to measure the expression level of ICAM-1 mRNAs and Southern blot hybridization to analyze the integration of human T-cell-leukemia virus type 1 (HTLV-1) provirus were done. The levels of ICAM-1 mRNA expression of ATL cells were generally higher than those of T-lymphoid cells. However, ILT-mat cells and ATL16T(-) cells, although they were ATL cells, showed rather low surface ICAM-1 expression and ICAM-1 mRNA expression. Southern blot hybridization showed that only two and four bands were found in ILT-mat and ATL16T(-) cells, respectively, whereas > 10 bands were detected in other ATL cells. These results suggest that monoclonal integration of HTLV-1 provirus to the genome of T cell, especially the number of integration sites, is one of the factors for induction of ICAM-1 molecules. PMID- 9219060 TI - Enzyme immunoassays for specific IgG and IgE antibodies to Pichia pastoris components in normal humans. AB - We developed enzyme immunoassays for human anti-Pichia pastoris components (PPC) IgG and anti-PPC IgE antibody titers. Anti-PPC IgG antibody assay were performed using antigen-coated plate and anti-human IgG peroxidase conjugate. The intra- and interassay coefficients of variation (CV) of anti-PPC IgG antibody were 1.83 2.51% and 1.97-2.76%, respectively. The anti-PPC IgE antibody assay was performed using an anti-IgE monoclonal antibody-coated plate, biotin-labeled PPC and avidin labeled peroxidase, which was not subject to interference by the high titer of anti-PPC IgG antibody. The intra- and interassay CV were 3.83-5.34% and 3.56 5.84%, respectively. We determined and compared anti-PPC IgG antibody titers in the 40 normal individuals. We confirmed that a high titer of anti-PPC IgG antibody is contained in all normal human sera and that these antibodies are directed primarily to mannan by immunoblotting analysis. The ratio of the maximum to minimum anti-PPC IgG antibody titers in normal individuals was > 8,000. Anti PPC IgG antibody titers did not correlate with the age. However, we did not detect anti-PPC IgE antibody in normal individuals. PMID- 9219059 TI - Temporal variability in immunological parameters: peripheral blood mononuclear cell subsets, serum immunoglobulins, and soluble markers of immune system activation. AB - T-cell subsets and soluble factors of immune system activation are increasingly used as biologic markers of disease and predictors of disease progression. For example, changes in CD4 cells and CD4:CD8 ratio, sIL-2R, B2M, neopterin, and IgA have been used in predicting AIDS onset and progression. We examined the temporal variability of T-cell subsets, monocytes, natural killer cells, B cells, immunoglobulins, soluble interleukin-2 receptor (sIL-2R), neopterin, and beta-2 microglobulin (B2M) among 135 adults tested at two time points approximately 3 months apart. The purpose of the study was two-fold: (1) to assess the stability of these measures at two points in time, and (2) to investigate which parameters tend to track together over time, i.e., show significant longitudinal correlation. Mean population values for these immunologic parameters remained remarkably stable over the 3-month period. However, individual subjects exhibited significant temporal variability for many parameters. Unlike observations in patients with AIDS, changes in immunoglobulins and other soluble factors were not significantly correlated with changes in cellular subsets over the same period. However, change in B2M was correlated with change in neopterin (r = .35, p < or = .0001), and change in IgA was correlated with changes in IgG and IgM (r = .44, r = .54, P < or = .001 for both). Characterizing this temporal variability in a healthy population provides important information for researchers applying these tests in clinical and epidemiological studies. PMID- 9219061 TI - Rapid diagnosis of acute eosinophilic pneumonia (AEP) in a patient with respiratory failure using bronchoalveolar lavage (BAL) with calcofluor white (CW) staining. AB - A diagnosis of exclusion, acute eosinophilic pneumonia (AEP) is an acute febrile illness with respiratory impairment, diffuse pulmonary infiltrates, and bronchoalveolar lavage (BAL) fluid eosinophilia. Whether pulmonary eosinophilia in AEP is primary or secondary remains undetermined. We report here a 22-year-old auto mechanic with severe AEP and acute respiratory failure who required intubation and ventilatory support. The patient's bronchoalveolar lavage (BAL) fluid was analyzed using cultures, cytology, Wright/Giemsa, Gram, Gomori methenamine-silver (GMS), and calcofluor white (CW) stains (1). Despite extensive evaluation, no infectious etiology was found. CW staining helped us rapidly to exclude Pneumocystis carinii or fungal infection and to focus attention toward the diagnosis of AEP. Transbronchial biopsy was unnecessary and supportive therapy without systemic glucocorticoids was followed by recovery within a few weeks. In this case, bronchoalveolar lavage with CW staining was of great assistance in the rapid diagnosis and initial management of AEP. Our literature review found no prior article using CW staining for evaluation of AEP. PMID- 9219062 TI - Hydroxyurea-induced denaturation of normal and sickle cell hemoglobins in vitro. AB - Use of hydroxyurea (HU) to treat sickle cell disease is usually associated with increments in fetal hemoglobin (Hb F) production; however, in vitro studies show that HU may also induce hemoglobin denaturation. Whole blood samples from Hb AA, Hb AS, and Hb SS patients were treated in vitro with 100, 150, 200, 250, and 300 micrograms/mL HU, incubated at 30 degrees C for up to 12 days, and analyzed by high-performance liquid chromatography (HPLC). Hb AA levels show decrements of 91 to 14% with 100 micrograms/mL and 89 to 4% with 150 micrograms/mL after 12 days; 86 to 2% with 200 micrograms/mL after 10 days; 86 to 8% with 250 and 300 micrograms/mL after 8 days. Similar treatment and incubation times for Hb AS whole blood demonstrate that HU equally degrades the A and S components of Hb AS. A comparable approach for Hb SS whole blood samples, using a 300 micrograms/mL HU treatment, showed a hemoglobin denaturing pattern that went from 93% to 1% after 12 days. Globin chain analysis of these samples by reverse-phase HPLC showed that the denaturing effects occur mostly on the beta-globin chain. PMID- 9219063 TI - Evaluation of the second generation IMx Toxo IgG antibody assay for detection of antibodies to Toxoplasma gondii in human sera. AB - For an evaluation of the Abbott IMx Toxo IgG second generation, antibodies to Toxoplasma gondii were detected by Abbott IMx Toxo IgG and IgM, Vidas Toxo IgG and Toxo IgM (bioMerieux, France) with immunofluorescence assay verified by the dye-test for IgG, and immunosorbent agglutination assay (ISAGA) for IgM as references. The study included 507 serum samples collected over one month in two laboratories, 32 samples from HIV-infected patients, and 70 serial samples from 23 women surveyed for seroconversion or persistent IgM. After exclusion of nine equivocal results from the 507 samples, the sensitivity and specificity, respectively, were 100% (156/156) and 100% (342/342) for the IMx Toxo IgG and 98.1% (153/156) and 100% (342/342) for the Vidas Toxo IgG. Of the 32 HIV-infected patient samples, 7 gave false positive results with IMx Toxo IgG. This was because the samples had been heated. In 5 of the 70 serial samples. IMx Toxo IgG gave positive results earlier than Vidas Toxo IgG and in two cases earlier than IgM antibody assays. In this study IMx Toxo IgG second generation showed an increase in sensitivity and specificity in comparison with data reported previously for the first generation. PMID- 9219064 TI - Simultaneous analysis of microheterogeneity of immunoglobulins and serum protein fraction using high-voltage isoelectric focusing on six cellulose acetate membranes. AB - A systematic detection method for the single performance of cellulose acetate (CA) membrane isoelectric focusing to detect six different types of information on protein abnormalities was developed. High-voltage isoelectric focusing was carried out on six layers of CA membrane using a thermoelectric cooling apparatus. After electrophoresis, the proteins on the top, the third, the fourth, the fifth, and the bottom CA membrane were transferred to a polyvinylidene difluoride (PVDF) membrane by a simple contact printing procedure to detect IgM, kappa-chain, lambda-chain, IgA, and IgG, respectively. Each PVDF membrane revealed the microheterogeneity of these immunoglobulins using specified anti serum and enzyme immunostaining. The second CA membrane was stained with Coomassie brilliant blue G250 to detect serum protein patterns. All stained membranes showed clear electrophoretic patterns of immunoglobulin microheterogeneity. By our method, immunoglobulin abnormalities in serum could be screened out using six different types of information obtained simultaneously. PMID- 9219066 TI - Unusual interference from primary collection tube in a high-performance liquid chromatography assay of amiodarone. AB - We describe an unusual interference in our routine high-performance liquid chromatography (HPLC) assay of amiodarone and its active metabolite, desethylamiodarone, used to quantify the parent drug and the active metabolite in serum from the primary sample collection tube. The interfering peak had a retention time very similar to that of the authentic desethylamiodarone. Substitution of Corvac tubes with Vacutainer tubes for the collection and transportation of serum samples eliminated the source of interference. We routinely suggest the use of Vacutainer collection tubes for obtaining blood samples of cardiac patients undergoing amiodarone therapy. PMID- 9219065 TI - Molecular diagnosis of pancreas carcinoma. AB - Cellular protooncogenes, tumor suppressor genes (antioncogenes), and DNA mismatch repair mutators are generally the key molecular genetic biomarkers undergoing alterations during carcinogenesis, i.e., activation of oncogenes, inactivation of tumor suppressors, and DNA mismatch repair gene defects are essential events in cancer causation. In pancreas cancer, high incidence of oncogene K-ras point mutations at the codon 12th is associated with premalignant and malignant transformation. Mutation in p53 tumor suppressor is also detected in pancreas adenocarcinoma. Concurrent loss of p53 and K-ras function may contribute to the clinical aggressiveness of pancreas cancer. Microsatellite instability and DNA mismatch repair defects may represent new mutator phenotype for pancreas carcinogenesis. Mutation of cell cycle regulators, such as inhibitor of CDK4 or p16 tumor suppressor gene, is a new molecular event in pancreas cancer. Mutation of cyclin-dependent kinases also may be involved in pancreas carcinogenesis. Loss or mutation of a new candidate tumor suppressor, DPC4 (deleted in pancreas carcinoma locus 4), is reported in pancreas cancer. The protein products of these gene mutations are potential tumor antigens, thus genotype expression can be detected by phenotype. Most of these emerging molecular genetic biomarkers are associated with regulation of cell growth and recognition, as well as gene expression, and may offer new insight into the cellular precursors to and genesis of pancreas cancer. PMID- 9219067 TI - Differentiation and resolution of erythrocyte and muscle adenylate kinase activities in serum by electrophoresis. AB - Adenylate kinase activity originating from erythrocytes has been shown to be distinct from muscle adenylate kinase or myokinase activity, until now considered to be identical enzyme activities. The two activities can be differentiated by electrophoretic fractionation, thus making it possible to quantify the erythrocyte adenylate kinase activity present in serum. PMID- 9219068 TI - A comparison of the effects of four therapy procedures on concentration and responsiveness in people with profound learning disabilities. AB - This paper is an investigation into the efficacy of four therapeutic treatment procedures increasingly used with people with profound learning disabilities: snoezelen, hand massage/aromatherapy, relaxation, and active therapy (a bouncy castle). In particular, the effects of these procedures on concentration and responsiveness were examined. Eight subjects with profound learning disabilities took part in the study and each subject received each of the treatments. To assess the effects of the treatments, simple concentration tasks were administered and the subjects' responsiveness to each treatment was rated by independent observers. The results suggest that both snoezelen and relaxation had a positive effect on concentration and seemed to be the most enjoyable therapies for clients, whereas hand massage/aromatherapy and active therapy had no or even negative effects on concentration and appeared less enjoyable. PMID- 9219069 TI - Issues for consideration in dihydropteridine reductase (DHPR) deficiency: a variant form of hyperphenylalaninaemia. AB - An adult male with intellectual disabilities demonstrated deterioration in many skills over a number of years and an increase in his temper outbursts was also reported. At 18 years of age, he had been diagnosed as having dihydropteridine reductase (DHPR) deficiency, but treatment had proved unsuccessful in the short term and was discontinued. Dihydropteridine reductase deficiency is a recessively inherited disorder of the amino acid metabolism resulting in a deficiency of tetrahydrobiopterin, an essential cofactor for phenylalanine, tyrosine and tryptophan metabolism. This causes a severe deficiency of neurotransmitters in the brain. Following further neurological examinations, treatment for the subject was recommenced at the age of 30 years. Few reports of late-diagnosis DHPR have been documented. This paper outlines one case report of DHPR, highlighting the importance of diagnosis, medical treatment and nursing care. PMID- 9219070 TI - Sexual abuse perpetrated by men with intellectual disabilities: a comparative study. AB - This paper compares cases of sexual abuse of adults with intellectual disabilities, reported across the South East of England, which were perpetrated by men with intellectual disabilities, with those committed by other male perpetrators. The comparison provides some support for the findings of other studies, which have suggested that men with intellectual disabilities offend against more male victims than non-disabled sex offenders and that their offences are somewhat less serious, but otherwise indicates common patterns of abusive behaviour across this divide but differential service responses and support for victims. So called 'peer abuse' is a widespread problem which service agencies have failed to address: repeated offences are frequent and lack of appropriate intervention is the norm. PMID- 9219071 TI - 'What do you think?': a qualitative approach to evaluating individual planning services. AB - An established individual planning service was evaluated using a service-user centred approach, which looked at the extent to which people are involved in the process and their understanding of its nature and function. Responses recorded during interviews with service users, their keyworkers and some family members are complemented with data collected from participant observation of individual planning meetings. The findings suggest that the majority of service users able to speak for themselves who were interviewed have a good understanding of the planning process and find it a positive experience; this is supported by participant observation data. Those speaking on behalf of people unable to speak for themselves are unclear as to how much understanding the group has of the process. Observation data suggest that people needing others to speak on their behalf are excluded from discussion during meetings more often than they are included. Recommendations are made for developing the service and providing additional support for keyworkers, building upon the considerable progress made during the service's inception. PMID- 9219072 TI - Prevalence study of the randomized controlled trials in the Journal of Intellectual Disability Research: 1957-1994. AB - Systematic reviews of care are increasingly potent guides to clinical practice, and it is important that all relevant randomized controlled trials (RCTs) are identified by those within the speciality of learning disability who produce such works. All RCTs in the Journal of Intellectual Disability Research and its predecessor were identified by hand-searching (1957-1994), and the frequency, origin, intervention and quality of reporting of randomization were described. Electronic searches for the trials were undertaken for the years 1974-1994, and the quality of indexing was inspected and tested. These electronic searches were then compared to a 'gold standard' search. Fifty-six RCTs were identified. None contained the world 'randomized' in the title and only nine mentioned it in the abstract. Out of the 37 RCTs published between 1974 and 1994, 36 are in PsycLIT and 37 in MEDLINE. One MEDLINE record contained the wrong abstract. The methodological phrases used in the electronic records were poor, and thus, the precision of electronic searches, using both databases, was low. This international journal contains many relevant RCTs from around the world, involving several types of interventions. Unfortunately, these trials cannot be readily accessed electronically using methodological phrases designed to find RCTs. Improved quality of indexing would facilitate identification of RCTs and their dissemination. PMID- 9219073 TI - Schizophrenia in people with intellectual disability: the role of pregnancy and birth complications. AB - The literature suggests that mental illness is more common in people with intellectual disability than in the general population. Having reviewed the literature, Turner (1989) [Psychological Medicine 19, 301-14] suggested that about 3% of people with intellectual disability also have schizophrenia. As pregnancy and birth complications (PBCs) occur more commonly in people with intellectual disability than in the general population and are also implicated in the aetiology of schizophrenia, it is possible that these conditions share a common aetiology. This study reports on the occurrence of PBCs in those people with intellectual disability who develop schizophrenia. Fifty people with intellectual disability and schizophrenia were matched for age, sex, degree of intellectual disability and presence of epilepsy with a control group who did not suffer from schizophrenia or a schizophreniform psychosis. The obstetric history was obtained and events rated on a scale specifically designed for this study. This PBCs scale consists of six sub-scales covering areas of general maternal health, pregnancy, delivery, medication in labour, total medication score and neonatal score, as well as an overall total score. The study found that people with intellectual disability who develop schizophrenia have significantly higher rates of PBCs than controls. All of the sub-scales on the PBCs scale were significantly higher in people with schizophrenia, with the exception of the medication scales. Only five out of the 50 people with schizophrenia had not had a major obstetric complication, compared to 13 subjects from the control group. A number of abnormalities were specifically higher in people who later developed schizophrenia. These included: abnormally long or short labour; maternal episiotomy; maternal preeclamptic toxaemia; induction of labour; dysmaturity; maternal smoking in pregnancy; and a delay in neonatal crying. The results suggest that PBCs are important in the aetiology of schizophrenia in people with intellectual disability. PMID- 9219074 TI - Mortality and avoidable death in people with severe self-injurious behaviour: results of a Dutch study. AB - Mortality and avoidable death was studied in a cohort of 1168 people with severe self-injurious behaviour (SIB) in the Netherlands. Fifty-seven people died over a 5-year period (1990-1995). The observed mortality in the cohort studied was higher than the expected mortality. The age-specific mortality was highest in the 30-39-year-old age-group. Diseases of the respiratory system were found to be the most prevalent cause of death, followed by diseases of the nervous system and sensory organs. In six people (12%), the general practitioner and staff member considered SIB to be related to death. The causes of death were thought to be avoidable in two cases. The results are discussed in terms of data collection and avoidability of death. PMID- 9219075 TI - Staff strategies and explanations for intervening with challenging behaviours: a replication in a community sample. AB - Carers' beliefs about challenging behaviours may partially determine their behavioural responses to them. The present study replicated previous work on the beliefs of institution staff and their explanations about interventions for challenging behaviours (Hastings 1996) with a sample of 56 community staff. Many immediate intervention strategies, and the staff motivation for these choices, were in conflict with behavioural approaches to challenging behaviour and would be considered counter-habilitative from this perspective. Staff were able to describe appropriate longer-term interventions. These basic findings confirmed those of previous research with institution staff. However, tentative comparisons suggested that community staff were more likely than institution staff (from previous research) to describe interventions involving the building of relationships with service users and the identification of the underlying causes of the behaviours. PMID- 9219076 TI - Early menopause in women with Down's syndrome. AB - We used the AAMR's Adaptive Behavior Scale to ascertain current menstrual status in a population-based sample of 157 women with Down's syndrome (DS) and 187 women with other intellectual disability, all 40 years of age or older. The age adjusted likelihood of menopause was twice as high in women with DS syndrome as in women with other intellectual disability (OR = 2.3; 95% CI = 1.1-4.9). Treated thyroid conditions did not influence menstrual status and did not modify the relationship between DS and menstrual status. These findings support the hypothesis that women with DS experience menopause at an earlier age and that this may be associated with accelerated aging. PMID- 9219077 TI - Paroxetine in depressed adolescents with intellectual disability: an open label study. AB - The aim of this study was to evaluate the efficacy and side-effects of paroxetine treatment in adolescents with mild intellectual disability and major depressive disorder (MDD). Seven adolescents (14.7-18.4 years of age) were treated with paroxetine (dosage 20-40 mg day-1). Clinical changes were assessed at the beginning of the pharmacological treatment and after 9 weeks utilizing the DSM-IV diagnostic criteria and the Montgomery-Asberg Depression rating Scale (MADRS). Four out of the seven subjects did not fulfil the DSM-IV diagnostic criteria after the 9-week treatment. The mean decrease in the total score on the MADRS was significant (41%). Some items of the MADRS showed significant improvement: inner tension (66%); lassitude (55%); apparent sadness (53%); inability to feel (44%); and reported sadness (43%). Three subjects showed sedation, two subjects gastrointestinal complaints and one subject insomnia; all these symptoms were transitory and not severe. No behavioural activation was evident. This preliminary, uncontrolled study of a few cases suggests that adolescents with intellectual disability and MDD may respond to paroxetine, and that adverse side effects are mild. PMID- 9219078 TI - Adverse effects of summer amongst people with learning disabilities: neuroleptic malignant syndrome. AB - The existing literature regarding neuroleptic malignant syndrome (NMS) amongst people with learning disabilities is limited. We describe three case reports of people with learning disabilities from the same geographical area who developed NMS within 3 weeks of each other. This was during the hottest part of a very hot summer. The presentation of NMS is discussed. The importance of carers attending to adequate hydration for people with learning disabilities and poor communication skills during the summer months is stressed. PMID- 9219079 TI - Behaviour disorder in an adolescent with terminal deletion of chromosome 3. AB - Individuals with terminal deletion of the long arm of chromosome 3 are rare and survival into adulthood has not been previously reported. A 15-year-old with this condition was studied and the difficulties in management of the manifest behaviours are described. This chromosomal abnormality may be associated with mental and functional deterioration as well as severe self-injurious behaviour, commencing in early childhood. PMID- 9219080 TI - The role of anesthesia in the advancement of surgical technique. PMID- 9219081 TI - Surgical research specialist certification: it's time to expand the program. PMID- 9219082 TI - Cellular mechanisms of bone repair. AB - The extent of callus formation about a bone fracture depends on the rigidity of fracture fixation. The mechanism that converts the mechanical stimulus into the biologic response is unknown. On the basis of existing literature, an attempt has been made to define a model that explains this mechanobiologic transduction. Once integrity of the bone has been disrupted, a sequence of biochemical and cellular events commences that induces inflammatory reactions. Messengers (e.g., metabolites of the clotting or complement system, eicosanoids, or growth factors) are released or activated. They control the migration, proliferation, and protein synthesis of cells that are essential for angiogenesis and connective tissue formation. The key component in this inflammatory sequence seems to be the macrophage. Growth factors (e.g., released by macrophages) stimulate endothelial cells to form capillaries and mesenchymal cells to synthesize their matrix. In mechanically neutral areas, the fracture cavity is revascularized and osteoblasts proliferate and form bone. In mechanically instable fracture areas, spreading capillaries are disrupted by shear forces. In these areas, therefore, the milieu becomes hypoxic again. This milieu seems to support the differentiation of chondrocytes that stabilize the fracture by cartilage formation. If the strength of repair tissue is surpassed, the disrupture of the repair tissue triggers the mechanisms of inflammation again and additional cells immigrate and proliferate. Their protein synthesis increases repair callus. The increase of callus formation, however, stops when the tissue is capable of resisting motion. Links to the callus formation in osteitis are shown. PMID- 9219083 TI - Platelet activating factor and thromboxane B2 production after cardiopulmonary bypass. AB - Platelet-activating factor (PAF) is believed to have a central role in the pathogenesis of ischemia-reperfusion injury. The purpose of this study was to investigate the relationships among production of PAF, thromboxane B2 (T alpha B2), and cardiopulmonary sequelae in patients undergoing coronary artery bypass graft surgery (CABGS). Venous blood from nine patients (eight men, one woman) undergoing scheduled CABGS, was sampled from central venous catheters before anesthetic induction, pre-cardiopulmonary bypass (CPB), 10 and 30 minutes post CPB, 10 and 30 minutes post-aortic declamping, and 120 minutes and 24 hours after the conclusion of CPB. Plasma levels of PAF and T alpha B2 were determined by radioimmunoassay kits (Amersham Canada Ltd.). PAF and T alpha B2 were significantly different between high-risk patients (group I; Canadian Cardiovascular Society class 3-4; n = 4) and low-risk patients (group II; Canadian Cardiovascular Society class 2; n = 5): group I PAF = 960 pg/mL, group II PAF = 159 pg/mL (P = 0.0029); group I T alpha B2 = 320 pg/mL, group II T alpha B2 = 229 pg/mL (P = 0.0262). Group I PAF was significantly greater than group II PAF: before CPB = 825 pg/mL (group I), 138 pg/mL (group II); during initiation of CPB = 1600-2015 pg/mL (group I), 158-200 pg/mL (group II) (P = 0.0143). Correlations between duration of CPB and peak PAF (r = 0.7049, P = 0.0339), peak PAF level, and time to extubation (r = 0.8863, P = 0.0079) were significant. PAF levels were different in patients requiring postoperative epinephrine (2124 +/- 700 pg/mL) or dopamine (667 +/- 266 pg/mL) (P = 0.0042). The production of PAF is determined by preoperative patient characteristics and duration of CPB. Increased levels of PAF in patients with unstable angina, left main coronary artery disease, or recent myocardial infarction are associated with the need for increased inotropy and prolonged ventilatory support following CABGS. The degree of PAF production during CPB may be a factor in postoperative instability in high risk patients. PMID- 9219084 TI - Role of organized intestinal lymphoid aggregates in intestinal regeneration. AB - Intestinal lymphoid tissue has a complex interrelationship with the epithelium. The epithelia of intestinal crypts associated with lymphoid aggregates have an increased proliferation rate. In the present study, the authors tested the hypothesis that organized intestinal lymphoid tissue (Peyer's patches) enhances intestinal regeneration by studying this process with and without an adjacent Peyer's patch. Forty adult male Sprague-Dawley rats had full-thickness ileal defects patched with cecal serosa to allow regeneration of ileal mucosa. Control animals (group I) had the patch constructed adjacent to a Peyer's patch, whereas this Peyer's patch was excised in group II. Intestinal regeneration in both groups was evaluated on the third, fifth, seventh, and ninth days after operation. During the early phase of regeneration, both epithelial cell proliferation and migration were decreased in the patched defect after excision of the Peyer's patch. Crypt cell production rate in the adjacent normal mucosa also was decreased after excision of the Peyer's patch. Excision of the Peyer's patch resulted in less well-developed crypts and villi. Wound contraction, however, was greater in the intestinal defect adjacent to the Peyer's patch until day 7. In conclusion, Peyer's patches have a facilitative effect on the healing of intestinal wounds by promoting both epithelial cell migration on the defect and epithelial cell proliferation in the crypts adjacent to the wound and by decreasing the rate of wound contraction. These findings support a role for intestinal lymphoid tissue in the regulation of epithelial cell maintenance. PMID- 9219085 TI - A porcine model of gastroesophageal reflux. AB - The exact mechanism by which the lower esophageal sphincter (LES) prevents gastroesophageal reflux disease (GERD) remains controversial. The two main reasons for this controversy are the lack of a good animal model and the only recent technologic advances in this field. To date, no animal models exist that use 24-hour pH studies, manometry, and endoscopy in evaluating GERD. The purpose of this study is to create a good model of GERD that uses this technology and to delineate the true nature of the LES more clearly. Recently, three-dimensional vector volume studies were performed and through detailed, cadaveric, anatomic studies, the vector volume profiles were correlated to distinct groups of muscle fibers at the gastroesophageal junction in humans. In this study, an anatomic analysis of the porcine gastroesophageal junction revealed an arrangement of muscle fibers similar to that in humans. A myectomy of the porcine oblique fibers was performed in nine piglets weighing 10 to 12 kg. The authors were able to demonstrate manometric attenuation of the porcine LES, gastroesophageal reflux by ambulatory 24-hour pH monitoring (P < 0.0025), and the creation of GERD by documenting esophagitis grossly and histologically. In conclusion, this model of GERD works and lends further support to the notion that the LES may be a discrete anatomic entity. PMID- 9219086 TI - A rat model for the evaluation of small-caliber vascular grafts. AB - This article describes the development of a new experimental model using rats for the evaluation of small-caliber vascular grafts. By modifying heterotopic heart transplantation, two 1.5- to 2.0-cm long vascular prostheses were interposed between a syngeneic donor heart and the recipient abdominal vessels in the form of vascular bridges. Once blood flow through the vascular grafts was reestablished, the donor heart resumed normal beating. The status of the vascular grafts could be easily monitored by palpation. Occlusion of the grafts stopped donor heart beating without affecting survival of the animals. Once the surgical method was mastered, the postoperative mortality was approximately 10%, and the total procedure took less than 2 hours. Although microvascular surgical technique and equipment are required, this model has several advantages, including easy detection of thrombotic occlusion of the grafts, the use of small animals of defined genetic background, the absence of effect of graft occlusion on the recipient's life, and possible repeated operation on the same animal. PMID- 9219087 TI - Arthroplasty in the goat hip. AB - The goat is a reliable and practical animal model for study of the hip. The authors describe a safe and effective operative approach to the goat hip. PMID- 9219088 TI - Modified carotid artery transposition for repetitive arterial blood gas sampling in large animals. AB - A modified surgical procedure for creation of a carotid loop for repeated percutaneous sampling of arterial blood gas was performed on 8 Holstein heifers and 14 horses. This approach permitted sampling of blood gas via fine-needle aspiration (one to three times daily) and/or catheterization for extended periods. It offers several advantages over previously reported techniques, including greater accessibility, absence of postoperative complications such as hematoma formation, and absence of foreign materials supporting the loop. PMID- 9219089 TI - Can the infusion of elastase in the abdominal aorta of the Yucatan miniature swine consistently produce experimental aneurysms? AB - The intraluminal elastase perfusion model has been proved to be potentially effective in producing abdominal aortic aneurysm in rodents, but it produced unpredictable results in larger animals. The purpose of this study was to explore the potential ability of such a model to produce experimental aneurysm consistently in the Yucatan miniature swine. Six Yucatan miniature swine received infusion with porcine elastase into an isolated segment of the infrarenal aorta. The excised arterial segments were examined macroscopically to assess the luminal surface characteristics and histologically to describe the different pathologic injuries induced by the elastase treatment on the intima, media, and adventitia of the arterial wall. Histologic examination revealed that the elastic network of the media was destroyed. In the first week after perfusion, altered smooth muscle cells were located in the intima and innermost layer of the media in juxtaposition with the occlusive thrombus. Infiltration of inflammatory cells was observed in these regions of elastic network and smooth muscle cell alterations. In the arterial segments of swine sustained for 3 weeks, a reduction of smooth muscle cells was noted in some areas. An important number of necrotic lesions was observed, and they were associated with the development of calcium deposits. Significant intimal hyperplasic reaction was identified at day 19 and again at day 21. However, no aneurysmal development was observed. This study constituted the first experiment with infusion of porcine elastase in the Yucatan miniature swine infrarenal aorta. The present experimental protocol induced important elastic network and smooth muscle cell alterations leading to severe necrotic lesions associated with calcium deposition, but it produced no aneurysmal dilatation. This model requires further testing to obtain a more complete degradation of the elastic network in both the media and adventitia and more significant collagenolysis without early thrombotic events. PMID- 9219090 TI - Photoisomerization of retinoic acids in ethanol under room light: a warning for cell biological study of geometrical isomers of retinoids. AB - Photoisomerization of all-trans-retinoic acid and the geometrical isomers [9-cis retinoic acid, 11-cis-retinoic acid, 13-cis-retinoic acid and 9,13-di-cis retinoic acid] in ethanol and their biological effects on F9 teratocarcinoma cells were analyzed. The rates of photoisomerization of the retinoic acids illuminated by fluorescent lamps (1,200 lx) increased in inverse proportion to their concentrations. When the ethanolic solution of all-trans-retinoic acid (10( 5) M) was kept under illuminated condition, the equilibrium mixture of the geometrical isomers of retinoic acid [all-trans-retinoic acid 25%, 9-cis-retinoic acid 10%, 11-cis-retinoic acid 10%, 13-cis-retinoic acid 30%, 9,13-di-cis retinoic acid 5% and unidentified compound 20%] formed at around 30 min. The apparent velocity of the photoisomerization was approximately 8 x 10(-7) mol/L.min. Equilibrium mixtures with similar compositions were obtained by the photoisomerization of other geometrical isomers. The geometrical isomers produced by the photoisomerization possessed significantly different biological effects in the induction of differentiation of F9 cells into parietal endoderm-like cells: activities of 9-cis-retinoic acid (ED50, 8 x 10(-7) M), 11-cis-retinoic acid (ED50, 8 x 10(-7) M), and 13-cis-retinoic acid (ED50, 8 x 10(-7) M) were approximately 1/10 of all-trans-retinoic acid (ED50, 8 x 10(-8) M), and activity of 9,13-di-cis-retinoic acid (ED50, 1 x 10(-5) M) was 1/100 of the level of all trans-retinoic acid. Further, the retinoic acids acted with each other additively on F9 cells. PMID- 9219091 TI - Purification and some properties of cobalamin-dependent methionine synthase from rat liver. AB - Cobalamin-dependent methionine synthase was purified from rat liver. The enzyme activity was separated into two peaks upon Mono-Q column chromatography. Peaks I and II of the enzyme, eluted in this order, were purified 18,000- and 44,000-fold in overall yields of 0.7 and 1.8%, respectively. Peak II methionine synthase, the major fraction, was homogeneous as judged by SDS-polyacrylamide gel electrophoresis. The enzyme was a large monomeric protein with an apparent molecular weight of 143,000 Da. Interconversion of the enzyme between the two peaks was not observed during purification procedures. The enzyme required S adenosylmethionine and a reducing system for activity. Apparent K(m) values of the peak II enzyme for 5-methyltetrahydrofolate and homocysteine were 75 and 1.7 microM, respectively. PMID- 9219092 TI - Study on optimal fat content in total parenteral nutrition in partially hepatectomized rats. AB - In order to investigate the optimal fat content for total parenteral nutrition (TPN) solutions, male Wistar rats were subjected to 70% hepatectomy and then placed, for five days, on one of five TPN regimens in which fat represented 0%, 10%, 20%, 30% and 40%, respectively, of the total calorie content. As serum triglyceride levels in the fat-treated groups were lower than those in the non treated normal rats, it was concluded that the administered fat was sufficiently hydrolyzed. The greater the fat content, the higher the regeneration rate of the remnant liver. Significant differences were found between the 0%-fat group and 20%-plus fat groups. Hepatic triglyceride level was significantly lower in the 20%-fat group. Hepatic protein level was significantly elevated in all fat treated groups. Serum phospholipids and total cholesterol due to the lecithin contained in fat emulsion were significantly elevated in the 30 and 40%-fat groups, indicating that fat content of 30 and 40% was excessive. The results suggest that TPN containing fat is superior to fat-free TPN for liver regeneration after partial hepatectomy, and that optimal fat content is estimated to be about 20% of total calorie content in the case of this fat emulsion. PMID- 9219093 TI - Effects of whey protein on calcium and bone metabolism in ovariectomized rats. AB - We studied the effects of whey protein (WP) from cow's milk on calcium and bone metabolism in ovariectomized (OVX) rats. Six-week-old female Sprague-Dawley rats were ovariectomized and fed a low-calcium diet (0.03% Ca, 0.3% P) for 4 weeks. The OVX rats were divided into three groups and subjected to two experiments: Exp. 1, Cont group (20% casein, 0.3% Ca), WP (1%) group (19% casein, 1% whey protein, 0.3% Ca) and Low-Ca group (20% casein, 0.03% Ca); and Exp. 2, Cont group (20% casein, 0.3% Ca), WP (1%) group (19% casein, 1% whey protein, 0.3% Ca) and WP (2%) group (18% casein, 2% whey protein, 0.3% Ca). The rats were fed each experimental diet for 4 weeks. The final body weight, weight gain, food intake and food efficiency showed no significant difference between the Cont and WP (1%, 2%) groups in Exps. 1 and 2. There were no significant differences in the calcium balance, serum ALP or serum calcitonin levels between the Cont and WP groups in Exp. 1. But the breaking energies of the WP (1%, 2%) groups were higher than those of the Cont groups in Exps. 1 and 2. As for the amount of calcium, phosphorus and magnesium in the femur, there were no significant differences between the Cont and WP (1%, 2%) groups; however, the amounts of total amino acids in the femur of the WP (1%, 2%) groups were higher than those of the Cont groups in Exps. 1 and 2. The amounts of proline and hydroxyproline in the femur of the WP (1%, 2%) groups were also higher than those of the Cont groups in Exps. 1 and 2. These data indicate that the milk whey protein influence in OVX rats is an increase in bone proteins such as collagen and enhanced bone-breaking energy. PMID- 9219094 TI - Suppressive action of docosahexaenoic acid enriched-Euglena on reduction of endothelium-dependent relaxation in stroke-prone spontaneously hypertensive rats (SHRSP). AB - Stroke-prone spontaneously hypertensive rats (SHRSP) were fed a diet containing docosahexaenoic acid (DHA)-enriched Euglena glacilis (DHA-Euglena) as the protein source from 5 weeks of age. The effects on endothelial functions were investigated by perfusion experimentation using mesenteric vasculature, and compared with the effects of antihypertensive drugs. (1) At 13 weeks of age, SHRSP fed the DHA-Euglena diet showed an average blood pressure of 220 mmHg, which was 20 mmHg lower (p < 0.05) than that in the control group, while SHRSP of the captopril-treated group (angiotensin I converting enzyme inhibitor: 200 mg/L drinking water) and hydralazine-treated group (vasodilator: 60 mg/L drinking water) showed marked hypotensive effects with blood pressures of 150-160 mmHg and 180-190 mmHg, respectively. (2) The constriction response to norepinephrine (NE) was lower (p < 0.01) in the mesenteric vasculature isolated from the DHA-Euglena treated SHRSP than in that from the control group. (3) When the mesenteric vasculature isolated from 13-week-old SHRSP fed the DHA-Euglena diet was perfused with an acetylcholine solution (10(-6) M) in the presence of NE (8 x 10(-6) M), the relaxation rate was 81%, which was higher (p < 0.01) than that in the control group (61%). Among the antihypertensive-treated groups, the captopril-treated group gave nearly the same relaxation rate as the DHA-Euglena diet group, while the hydralazine-treated group indicated a slightly lower rate (65%). At 18 weeks of age, the endothelium-dependent relaxation of SHRSP in the control group was further reduced (28%), but in both the DHA-Euglena diet group and antihypertensive-treated groups, the relaxation rates were not substantially different from the levels at 13 weeks of age. Reduction of the endothelium function in SHRSP occurs due to aging and blood pressure elevation. However, by improving nutritional conditions by the feeding of a DHA-Euglena diet, the endothelial functions were protected without a fall in blood pressure unlike antihypertensive drugs. It is hence considered that nutritional improvement helps maintain a sound architecture for the vascular wall, thereby leading to the suppression and delay of onset of cerebrovascular diseases, and subsequently to the prolongation of life-span. PMID- 9219095 TI - A comparative study of high-fat diet containing fish oil or lard on blood glucose in genetically diabetic (db/db) mice. AB - The effects of high-fat diets containing fish oil or lard on blood glucose and plasma insulin after oral glucose loading were compared in genetically diabetic (db/db) mice, one of the animal models of non-insulin-dependent diabetes mellitus (NIDDM) with hyperinsulinemia. The blood glucose levels were significantly decreased in 27% of the mice fed high-fat diets containing 20% fish oil 30 and 60 min after the oral administration of glucose (both; p < 0.05). Conversely, the plasma insulin levels were significantly increased 30 min after the glucose loading as compared to 27% of the mice fed high-fat diets containing 20% lard (p < 0.01). In addition, a significant hypoglycemic effect was observed 60 min after the subcutaneous administration of insulin to mice on the fish oil diet (p < 0.05), whereas no effect was demonstrated in the case of those on the lard diet. The average body weight of the fish oil-treated mice was not significantly different from that of the lard-treated mice. The fish oil diet has a beneficial effect on glucose tolerance by increasing the insulin secretory capacity from pancreatic beta cells and also ameliorating insulin resistance. PMID- 9219096 TI - Change of tryptophan-niacin metabolism in D-galactosamine-induced liver injury in rat. AB - The change of tryptophan-niacin metabolism in D-galactosamine (D-galN) injected rats was investigated. Rats fed with niacin-free diets containing 40% casein for 11 days were injected with D-galN (0.8 g/kg body weight). The urinary excretions of nicotinamide and its metabolites, and the activity of liver alpha-amino-beta carboxymuconate-epsilon-semialdehyde decarboxylase (ACMSD) (EC 4.1.1.45), a key enzyme of tryptophan-niacin metabolism, were assayed. As the result, the urinary excretions of N1-methylnicotinamide (MNA), N1-methyl-2-pyridone-5-carboxamide (2 Py), N1-methyl-4-pyridone-3-carboxamide (4-Py) and their sum (nicotinamide+MNA+2 Py+4-Py) were higher in the D-galN-injected group than in the control group. Hepatic ACMSD activity in the D-galN-injected group was lower than that of the control group. These results suggest that the increase in urinary excretion of nicotinamide and its metabolites after the injection of D-galN is considered to be attributable to a decrease in liver ACMSD activity. PMID- 9219097 TI - Fumaric acid, anti-thrombin substance from Rhizopus javanicus. AB - An anti-thrombin substance (M2) was isolated from a culture broth of Rhizopus javanicus. Accumulation of M2 reached a maximum peak after 14 to 15 days of incubation and then decreased. The yield of M2 was 500 mg from 1 L of culture broth. M2 inhibited thrombin activity, and its 50% inhibition concentration in a reaction mixture containing 50 microL of 12.5 NIH unit/mL thrombin and 200 microL of 0.33% bovine fibrinogen was 63 microM. M2 had a specific activity for thrombin, but it was less responsive to plasmin, tissue-type plasminogen activator, urokinase, plasma kallikrein and glandular kallikrein. The structure of M2 was identified as fumaric acid by elementary analysis, FAB/MS, 1H-NMR, 13C NMR and IR spectra. PMID- 9219098 TI - Antioxidative mechanism and apoptosis induction by 3-hydroxyanthranilic acid, an antioxidant in Indonesian food Tempeh, in the human hepatoma-derived cell line, HuH-7. AB - Recently, a new potent antioxidant was isolated from Tempeh (a traditional fermented soybean food in Indonesia) and was identified as 3-hydroxyanthranilic acid (HAA). This study deals with the antioxidant mechanism of HAA under biological systems and the cytokilling function of HAA to human malignant cells. HAA eliminated free radicals and inhibited the formation of fatty acid hydroperoxide in vitro, suggesting that HAA would serve as an antioxidant in the initial reaction in lipid oxidation systems. Actually, HAA inhibited the formation of the dominant product of membrane lipids, 12-hydroxyeicosatetraenoic acid (12-HETE) at a high concentration, while HAA accelerated 12-HETE formation at a low concentration in mammalian tissue. HAA oxidized glutathione and inhibited superoxide dismutase in vitro. Furthermore, HAA inhibited cell growth and induced apoptosis to HuH-7, a human hepatoma-derived cell line. As long as HAA is taken as a component of Tempeh, and not in large doses as a chemical, it may possibly act as a prooxidant rather than an antioxidant in vivo. PMID- 9219099 TI - Effects of cabbage leaf protein concentrate on the serum and liver lipid concentrations in rats. AB - The effects of cabbage leaf protein concentrate (CLPC) on serum and liver lipid concentrations were determined in rats fed cholesterol-enriched and cholesterol free diets. In rats fed the cholesterol-enriched diet with CLPC, total cholesterol, triacylglycerol and phospholipid concentrations in both the serum and liver, as well as the atherogenic index diet were significantly lower than those of the rats fed a casein diet. A supplement of methionine to the CLPC diet raised serum HDL-cholesterol and body weight gain, indicating that the addition of methionine to the CLPC diet is not only available to improve the nutritive value of CLPC but also to lower the atherogenic index. In rats fed the cholesterol-free diet, the liver total cholesterol and triacylglycerol concentrations of the CLPC-fed rats also showed lower values than those of the casein-fed rats, however, the serum total cholesterol concentration of the CLPC fed rats did not differ from that of the casein-fed rats. PMID- 9219100 TI - The menopausal misnomer. PMID- 9219101 TI - Secular trends in menopause age. AB - The question of whether or not there has been a secular trend in age of menopause in Europe and the United States from the 1940s to the 1990s has intrigued many investigators. However, since 1978, no attempt has been made to update Flint's 1978 study, from 1948 to 1964, of menopause ages in Denmark, England, France, Germany and the United States. This paper will attempt to describe the problems of doing secular trend research. These include methodological problems such as the use of retrospective dates and population representations, as well as the factors that affect the natural biological age of this reproductive landmark. Suggestions are made for ways to standardize menopause age research cross culturally. These include having large enough populations studied to be statistically significant and then randomly selecting the same populations in specific areas of a country and studying them every 10 years or more; studying generations of women in same families in various areas of a particular country; or studying cohorts of women, both cross-sectionally and prospectively. An International Menopause Index for worldwide ages of menopause is also proposed. PMID- 9219102 TI - Menopause in different cultures. PMID- 9219103 TI - Medicalization of women's third age. AB - Medicalization usually refers to the process whereby the normal processes of pregnancy, childbirth, menstruation and menopause have been claimed and redefined by medicine. Rather than discussing medicalization and menopause in terms of the number of women taking hormones, or the percentage of physicians convinced they should prescribe them, this paper looks at the visual image of the menopausal woman as portrayed in the pharmaceutical literature and in the mass media. Unlike the depressed and sickly looking women shown in the pharmaceutical advertisements in the 1970s, this 1990s version of the menopausal woman is shown glowing with fitness, with well-maintained teeth, hair and skin, far too fit to break a hip, have a heart attack, or witness the slow destruction of their minds by Alzheimer's disease. This image is not to be confused with the reality of being a menopausal woman, yet the two are intimately intertwined, for the image determines how menopausal women see themselves and how they are seen in the wider society. The final section of the paper discusses how health is the new virtue for women as they age as each individual is held responsible for what happens to her body, particularly in terms of the decisions made at the time of menopause. PMID- 9219104 TI - The social construction of menopause as risk. AB - Menopause and postmenopause are increasingly defined as a period of health risks that require medical and pharmacological treatment. A social construction of menopause as risk is taking place in popular culture and in the medical field and implies a medicalization of women's lives. PMID- 9219105 TI - Well-being and the menopausal transition. AB - This paper examines the relationship between well-being, age, menopausal status, hormone levels and hot flashes. Data from the first 4 years of longitudinal observation from the Melbourne Women's Midlife Health project was utilized. This study involved a population-based sample of 405 women interviewed annually. Blood was taken during the follicular phase (if still menstruating) for estradiol, sex hormone binding globulin, follicle stimulating hormone and testosterone. A validated well-being scale was used. Positive affect increased with age while negative affect decreased with age but only in the postmenopausal category. Positive affect was significantly lower in the 2 years postmenopausal group but this effect of menopausal status did not remain when hot flashes were included in the analysis. Negative affect was highest in the 1-2 years postmenopausal group. Although hot flashes adversely affected negative moods, a significant effect of menopausal status remained. No direct association between any of the hormone levels and positive or negative affect scores was evident. In conclusion, this study found that well-being was decreased in the first 2 years after the first menstrual period but began to improve spontaneously after this time. PMID- 9219106 TI - Menopause and osteoporosis: theoretical aspects. Effects of pluriform practices for present day health care and for women. PMID- 9219107 TI - Menopause and cardiovascular disease. PMID- 9219108 TI - Menopause and the brain. AB - Estrogen may have a beneficial effect on the risk and course of Alzheimer's disease (AD) through several mechanisms, including improvement of cerebral blood flow, stimulation of the neuron, or gliacyte and interaction with genetic factors. In this paper, the therapeutic and etiologic research of the role of estrogen in cognitive function and dementia is reviewed. Findings to date are promising but far from conclusive. In therapeutic research, interpretation of studies is hampered by the small sizes of the studies and differences in methodology. Most etiological studies have been limited to retrospective studies in which the history of estrogen use was obtained from an informant. Follow-up studies conducted to date have yielded controversial results. Further research is needed to elucidate the role of estrogen in the pathogenesis and progression of dementia. Subjects genetically susceptible for AD may prove to be an important high-risk group to target in preventive, therapeutic and etiologic research. PMID- 9219109 TI - Hormones and sexuality in postmenopausal women: a psychophysiological study. AB - Sexual function, including vaginal atrophy, and hormonal status, were studied in 42 naturally postmenopausal women. Vaginal pulse amplitude and subjective sexual responses during self-induced erotic fantasy and during erotic films were compared with responses of a small number of premenopausal women. As predicted, vaginal atrophy was related to estrogens but not to complaints of vaginal dryness and dyspareunia. No significant relationship was found between hormones and sexual function. Unexpectedly, most of the few correlations that did reach significance involved prolactin. The fact that prolactin was negatively associated with sexual desire, sexual arousal and vaginal lubrication during sexual activity, suggests that psychosocial factors are more important than hormone levels in postmenopausal sexual function. Comparisons with a number of premenopausal women revealed that although postmenopausal women displayed lower vaginal pulse amplitude responses prior to erotic stimulation than the premenopausal women, this difference disappeared during subsequent erotic stimulation. We argued that this finding can be interpreted as being supportive of the notion that complaints of vaginal dryness and dyspareunia should not be attributed to vaginal atrophy associated with menopause. Rather, vaginal dryness and dyspareunia seem to reflect sexual arousal problems. PMID- 9219110 TI - Menopause, only for women? The social construction of menopause as an exclusively female condition. AB - Over the last three decades the menopause has continued to interest the medical profession, the pharmaceutical industry and the mass media. Although there exist many different views on the menopause, there is one common denominator. Menopause is depicted as an exclusively female condition. The medical discourse on menopause seems to exclude men. However, a closer look at the history of the medical sciences reveals that there have been and still are, attempts to classify symptoms of ageing men as male menopause or climacterium. Despite these attempts to put men on the menopausal agenda, most attention is focused on women. How can we understand this almost exclusive focus on female bodies? Why does there exist such an emphasis on the medicalization of the third age of women rather than of men? Maybe we might be inclined to think of a male conspiracy, as has been suggested by feminists: women take the pills, while men cash the bills. We might consider the enormous profits of the pharmaceutical industry. This paper is concerned with finding an alternative explanation for the almost exclusive attention for the female menopause. Based on historical data and more recent discussions in medical journals, the paper shows that the medicalization of the female menopause and the relative silence around the male climacterium can be understood in terms of the social and cultural processes that underly the classification of health problems as specific diseases. The imbalance in medical treatment of climacteric health problems in women and men is not simply rooted in biological sex differences, but can be ascribed to men's attitudes towards health problems and organizational infrastructures of the medical institutions. PMID- 9219111 TI - Prescribing of hormone therapy in menopause and postmenopause. AB - This article describes the use and prescribing of menopausal and postmenopausal hormone therapy (HT) in one example country, Finland, and the trends and levels of HT use in other western countries for comparison. Previously published studies were reviewed and reanalyzed, and some additional unpublished data from Finnish surveys were compiled. The use of HT increased in Finland up to 1994. In Finland the initiative for HT use came more often from physicians than women themselves, physicians valued HT more than women, women's period of use of HT was shorter than physicians' recommendations, women's reasons for using HT were usually to treat symptoms, but physicians considered HT also useful in the prevention of later diseases. Gynecologists were more favorable toward HT than other physicians. HT has become common in very different times in different countries, but with the exception of the US experience in the 1970s, the trend has been towards increasing use. One motivation to do surveys on physicians' prescribing or women's use of HT has been to facilitate HT use. The large variation in HT use may reflect the uncertainty concerning its true value. The reasons for the large scale prevention with HT have not been systematically studied, but it is likely due to various social and commercial forces. PMID- 9219112 TI - The marketing of estrogens for menopausal and postmenopausal women. AB - Menopausal and postmenopausal women are a targeted consumer group in the promotion of hormone drugs. The marketing of these pharmaceutical products aims directly at prescribing medical professionals and indirectly at women. The promotion seems intended to creating a collective consciousness that women over 40 need medical and pharmacological treatment. PMID- 9219113 TI - Hormone replacement therapy and the risk of reproductive cancers. PMID- 9219114 TI - Medicalization of menopause and public health. PMID- 9219115 TI - The menopause and the pharmaceutical industry. AB - Since 1975, pharmaceutical firms specializing in the endocrine field have made a vigorous attempt to develop a large market in estrogens for the long-term treatment of postmenopausal women. The success of the contraceptive 'pill' had demonstrated the commercial attractiveness of developing products for an essentially healthy population. The scientific challenge was to demonstrate the value of such treatment, e.g. in countering osteoporosis; another problem was the generally poor compliance observed when patients receive drugs other than for the relief of acute conditions. Techniques used included the selective emphasis to physicians of those research findings pointing to the value of hormonal replacement, the development of direct approaches to the public to stimulate interest in a treatment which might prolong life or delay some parts of the ageing process ('feminine forever'), and the creation of intermediary institutes to undertake selective research and influence opinion. PMID- 9219116 TI - Stress urinary incontinence. Laparoscopically assisted transvaginal needle suspension. AB - OBJECTIVE: To characterize laparoscopically assisted transvaginal needle suspension for stress urinary incontinence and describe the authors' initial experience with this approach. STUDY DESIGN: A retrospective analysis was performed on 19 cases of stress urinary incontinence treated with this technique. All patients were evaluated at least one year postoperatively. RESULTS: Laparoscopically assisted transvaginal needle suspension was successfully completed in all 19 cases. No procedure required conversion to laparotomy. No intraoperative morbidity attributable to prevesical dissection was noted. Average operating time for needle suspension was 25 minutes, and average blood loss was 50 mL. No postoperative infectious or hemorrhagic morbidity was noted. All patients were subjectively cured of incontinence one year postoperatively, although two patients experienced transient urinary retention, and one experienced transient detrusor instability within the first three postoperative months. CONCLUSION: This approach allows accurate, visually guided placement of suspension sutures and may induce extraperitoneal scarring similar to retropublic colposuspension. These factors may result in long-term cure rates comparable to those of retropublic colposuspension without the need for endoscopic suturing. PMID- 9219117 TI - Polycystic ovary syndrome. An autoimmune disease? AB - OBJECTIVE: To determine whether there is an autoimmune cause of polycystic ovary disease (PCOD) implicating antiovarian antibodies in the pathophysiology of the syndrome. STUDY DESIGN: This study examined 31 women diagnosed as having PCOD according to internationally recognized criteria and who attended our infertility clinic during 1994-1995. RESULTS: We could not confirm the findings of previous studies. Only one patient in our group with type II diabetes mellitus had antiovarian antibodies present in her serum. CONCLUSION: It seems that an autoimmune mechanism probably is not of prime importance in this multifaceted syndrome. PMID- 9219118 TI - ASCUS on cervical cytologic smears. Clinical significance. AB - OBJECTIVE: To determine the significance of the category atypical squamous cells of undetermined significance (ASCUS) in predicting the presence of underlying squamous intraepithelial lesion (SIL) and to determine the best follow-up method. STUDY DESIGN: Follow-up studies of all cervical cytologic smears with a diagnosis of ASCUS within 14 months were reviewed. RESULTS: SIL was diagnosed in 66.67% of ASCUS patients upon follow-up with biopsies with and without smears. SIL was diagnosed in 17.39% of ASCUS patients upon follow-up with smears only. CONCLUSION: Based on this study, ASCUS on smears serves as a good marker of underlying SIL. Follow-up studies with biopsies with and without smears appear to be more effective in detecting underlying low grade SIL than repeat smears only. PMID- 9219119 TI - Determining blood pressure in pregnancy. Positional hydrostatic effects. AB - OBJECTIVE: To evaluate positional hydrostatic effects on blood pressure determination during pregnancy. STUDY DESIGN: We studied 30 normotensive, pregnant women at 34-41 weeks of gestation. Blood pressures were taken in the sitting, left lateral, right lateral and supine positions with a two-minute stabilization period between positions. The bisacromial diameter was measured. Multivariate analysis of variance for repeated measures was used to evaluate the affect of position on blood pressure. RESULTS: Mean systolic pressure in the right arm was 2.6 mm Hg greater than that in the left arm (P < .05). There was no difference between the arms in diastolic blood pressure. Immediate blood pressure in the lower arm was no greater than in the higher arm in lateral positions, and there were no other significant positional effects. Observed blood pressures were significantly different than those theoretically expected on the basis of hydrostatic effects (P < .0001). CONCLUSION: Positional effects on blood pressure in the lateral positions do not appear immediately (within two minutes), indicating that hydrostatic pressure does not account for these changes. The well documented blood pressure reduction from longer duration in the lateral position does not appear to be an artifact of hydrostatic effect. Repositioning pregnant women in the supine position to have the cuff at the level of the heart is unnecessary and often undesirable when fetal perfusion is an important consideration. We suggest that American Heart Association blood pressure guidelines stating that all measurements be taken with the cuff at the level of the heart to avoid hydrostatic pressure change be revised for pregnancy. PMID- 9219120 TI - Hysteroscopy, hysterosalpingography and tubal ostial polyps in infertility patients. AB - OBJECTIVE: To compare the findings in infertility patients who underwent preoperative hysterosalpingography (HSG) followed by hysteroscopy and to determine the incidence of tubal ostial polyps, their HSG appearance and the results of hysteroscopic resection in our patient population. STUDY DESIGN: Sixty eight infertility patients were evaluated by HSG followed by hysteroscopy. HSG diagnoses were divided into groups: group 1, normal; group 2, bilateral tubal occlusion; group 3, unilateral tubal occlusion; group 4, filling defects; and group 5, abnormal cavity. HSG findings were compared to the hysteroscopy findings. For patients in whom tubal ostial polyps were found, the findings were described, including postsurgical interval to conception. RESULTS: The agreement rates were 90%, 50%, 69%, 73% and 71% for groups 1-5, respectively. The positive predictive value of an abnormal HSG was 65%, and the negative predictive value of a normal HSG was 90%. Six of 68 patients (11.3%) had polyps at the fallopian tube ostium. Three of these patients (50%) had had the finding of proximal tubal occlusion on the ipsilateral side predicted by HSG; three had had normal HSGs. Four of the six conceived following polypectomy. The mean interval from surgery to conception was 4.5 months. CONCLUSION: HSG was a specific but not sensitive predictor of uterine pathology in our patient population. Tubal ostial polyps may occur in a significant proportion of infertility patients and can cause proximal tubal occlusion on HSG. Their possible contribution to infertility and clinical significance deserve further investigation. PMID- 9219121 TI - Ultrasonographic fetal cardiac measurement in isoimmunized pregnancies. AB - OBJECTIVE: To investigate the predictive value of the ultrasonographically measured fetal biventricular outer dimension (BVOD) in diastole in detecting neonatal anemia in pregnancies complicated by isoimmunization. STUDY DESIGN: The records of all patients evaluated for isoimmunization in pregnancy from January 1992 to December 1994 were reviewed retrospectively. The fetal BVOD had been measured with real-time-directed M-mode fetal echocardiography. The BVOD measurement was plotted on a nomogram (with reference to biparietal diameter) and a percentile value determined graphically from the nomogram. Neonatal outcome was obtained prospectively and by chart review. RESULTS: Sixty-three singleton fetuses from the study period who met entry criteria were identified. Anti-D sensitization represented 66% of cases of isoimmunization. Twenty (32%) fetuses required subsequent neonatal transfusion, and 43 (68%) did not. Seventeen fetuses (27%) had BVOD measurements greater than the 95th percentile, and 10 (59%) required subsequent transfusion. Infants in this group also had significantly lower hematocrits at birth (37.7 +/- 13.0% vs. 46.6 +/- 9.0%) and prolonged neonatal intensive care unit stay (10.7 +/- 10.0 vs. 4.7 +/- 3.6 days), respectively, when compared to patients with a BVOD measurement less than the 95th percentile. A BVOD 95th percentile threshold had a sensitivity, specificity and positive predictive value of 50%, 84% and 59%, respectively, in predicting the need for neonatal transfusion. CONCLUSION: In patients with isoimmunization, a BVOD measurement in the 95th percentile or greater was associated with a relatively high likelihood of neonatal anemia and transfusion. Although the measurement is not sufficiently sensitive to be used as a single parameter in predicting neonatal compromise in these patients, it can be a useful, noninvasive adjunct to the management of isoimmunized pregnancies. PMID- 9219122 TI - Cesarean section for suspected fetal distress. Does the decision-incision time make a difference? AB - OBJECTIVE: To compare perinatal outcomes in patients at term (37 weeks) in whom the decision-incision time for cesarean delivery was due to suspected fetal distress. STUDY DESIGN: All parturients who underwent cesarean delivery primarily for possible fetal distress during a three-year period were identified retrospectively. Student's t test and the chi 2 test were utilized, and P < .05 was considered significant. A regression analysis of decision-incision time and umbilical arterial pH was performed. RESULTS: From 1991 to 1993, 1.3% (117/9,137) of term laboring patients underwent emergency cesarean delivery for the primary indication of possible fetal distress. In 61 patients (52%) the decision-incision time was 30 minutes, while it exceeded 30 minutes in the remaining 56 women. The two patient groups were similar in maternal demographics, antepartum complications, oxytocin usage, thick meconium, type of abnormal fetal heart rate tracing prompting surgery, use of amnioinfusion (41% vs. 36%), general anesthesia (97% vs. 93%), mean birth weight and Apgar score < 7 at five minutes. Three adverse outcomes were observed more frequently in association with decision incision time > 30 minutes: (1) lower mean (+/-SD) umbilical arterial pH (7.16 +/ 0.15 vs. 7.26 +/- 0.06, P = .001), (2) pH < 7.00 (8/61 vs. 0/56, P = .005), and (3) admission to the neonatal intensive care unit (P = .008). When the incision was made longer than 30 minutes after the decision, there was no apparent adverse neonatal or infant outcome. CONCLUSION: Although a cesarean decision-incision time < or = 30 minutes is a desirable goal for the fetus possibly in distress, failure to achieve this goal is not associated with a measurable negative impact on newborn outcome. PMID- 9219123 TI - Prenatal determination of genotypes Kell and Cellano in at-risk pregnancies. AB - OBJECTIVE: To evaluate the accuracy of a DNA-based testing methodology in determining the KEL1 and KEL2 (Kell and Cellano) genotype of fetuses at risk for Kell or Cellano hemolytic disease. STUDY DESIGN: DNA was extracted from chorionic villus samples (CVS) or amniotic fluid (AF) cells, a portion of the Kell gene was amplified, the amplified product was cut with a restriction enzyme that recognizes the KEL1 nucleotide substitution, and the digested product was run on a polyacrylamide gel to separate the fragments. This analysis was routinely run on uncultured cells to provide rapid results. Testing of parental DNA was performed in conjunction with fetal analysis to ensure that their alleles were detectable with this DNA test. RESULTS: We determined the fetal KEL1 and KEL2 genotype in 1 CVS and 65 AF specimens. Forty-eight of them were determined to be KEL2, 17 were KEL1/2, and 1 was KEL1. Among the fetuses born to date, follow-up information was available on 14 of them, 11 KEL2 and 3 KEL1/2. In all 14 there was complete correlation between the DNA analysis and the serotype or clinical course. CONCLUSION: Determination of the fetal KEL1 and KEL2 genotype using this DNA-based method provides accurate and timely information that can aid the prenatal care of women sensitized to these Kell antigens. PMID- 9219125 TI - A new laparoscopic technique for interstitial pregnancy resection. A case report. AB - BACKGROUND: The interstitial gestation prevalence ranges from 1 in 2,500-5,000 live births, with a mortality rate of 2-2.5%. Total abdominal hysterectomy is usually offered to clinically stable patients over 35 years old. For younger women who desire to preserve their child-bearing capacity, interstitial pregnancy excision with a salpingo-oopherectomy is offered. The case presented here differed from the classic approach in that the adnexal structures were preserved, the products of conception were excised laparoscopically, and myometrium reconstruction was performed by suturing and tying with a laparoscopic technique. CASE: A 34-year-old multipara presented with genital tract bleeding and gradual onset, diffuse pelvic cramps progressively worsening and associated with nausea and vomiting. As expected, the regular, last menstrual period was 8 weeks prior to the patient's hospital admission. The serum beta-human chorionic gonadotropin level was 20,159 mIU/mL and the progesterone level 12.6 ng/mL. High-resolution transvaginal ultrasound revealed no intrauterine gestational sac. A 1.2-cm fluid collection was observed in the right uterine cornual area, and posterior cul-de sac fluid was present. A hysteroscopic evaluation was inconclusive, and the endometrium frozen section histologic study failed to confirm the presence of chorionic villi. The patient underwent diagnostic and operative laparoscopic excision of the interstitial pregnancy with adnexa preservation. CONCLUSION: Laparoscopic resection of interstitial pregnancy with the preservation of the adnexal structures was a safe alternative to laparotomy in this case. PMID- 9219126 TI - Acetaminophen poisoning in late pregnancy. A case report. AB - BACKGROUND: Acetaminophen poisoning is a major cause of hospital admission and has been extensively reviewed. Its occurrence in pregnant women has been reported seldom, and the prognosis has been good except for one case, in which the fetus died. We report on a case of acetaminophen poisoning that resulted in the death of both the mother and the infant. CASE: A 38-year-old woman whose pregnancy was at 31 weeks' gestational age was evaluated for treatment of an acetaminophen overdose. She was admitted more than 26 hours after taking 35 g of acetaminophen. An emergency cesarean section was performed one hour after admission because of acute fetal distress. A grossly normal, 1,620-g, female infant was delivered and had Apgar scores at 1, 5 and 10 minutes of 0, 0 and 1, respectively, despite the initiation of resuscitation immediately following delivery. Acidosis was noted in the mother during the operation; it was followed by acute hepatorenal failure 16 hours after admission. That resulted in the mother's death 40 hours after admission. The infant also died 34 hours after delivery. CONCLUSION: Delays in administering the antidote treatment, N-acetylcysteine, after acetaminophen intoxication significantly increase the risk of mortality in both the mother and infant. The development of acidosis carries a poor prognosis in such patients and may necessitate liver transplantation to save the life of the mother. PMID- 9219124 TI - Severe bleeding from a pregnancy tumor. A case report. AB - BACKGROUND: Hyperplastic gingivitis and gingival hyperplasia accompanying gestation have been termed "pregnancy gingivitis" and "pregnancy tumor." The condition is benign but, rarely, is complicated by severe bleeding that is difficult to manage. A single case of a pregnancy tumor is reported. CASE: A 28 year-old woman in the third trimester was evaluated for treatment of a pregnancy granuloma with recurrent episodes of severe bleeding for two weeks. Conservative management by firm pressure on gauze packs was applied to control the bleeding, but in vain. Induction of labor was conducted at 37 weeks partially because of term pregnancy and uncontrollable bleeding from the gingiva. An emergency cesarean section was done because of acute fetal distress during induction of labor. A healthy infant was delivered. The gingival bleeding stopped spontaneously five days afterwards. The patient was given thorough dental prophylaxis and oral hygiene instructions. The buccal granulomatous tumor was decreased in size four weeks postpartum. CONCLUSION: Careful oral dental hygiene, removal of dental plaque and debris, and use of soft toothbrushes are important during pregnancy to avoid occurrence of a pregnancy tumor. If uncontrolled bleeding occurs, management should be based on the individual condition and should range from supportive therapy--such as desiccation of bleeders; local, firm compression and oral hygiene to blood transfusion--as well as medication to accelerate fetal lung maturity or even termination of pregnancy to save the patient's life, as with treatment of uncontrollable eclampsia. PMID- 9219127 TI - Surgically correcting a vesicouterine fistula with a myouterine flap. A case report. AB - BACKGROUND: The incidence of vesicouterine fistula has been increasing, most probably secondary to a corresponding increase in the use of low segment cesarean section. CASE: A 37-year-old woman with a history of two cesarean sections, 14 years and 5 months earlier, presented with urge incontinence, cyclic hematuria and amenorrhea. Hysterosalpingography demonstrated contrast with the bladder and suggested a vesicouterine fistula. Following exploratory laparotomy and dissection of the bladder from the uterus, a fistula was seen connecting the anterior surface of the uterus and the posterosuperior aspect of the bladder. The fistula, with a cuff of uterus and bladder, was excised and the remaining defects repaired. In addition, a myouterine flap was raised to reinforce the repair. Upon follow-up the patient reported no difficulty in urination, complete urinary continence, normal menses and no hematuria. CONCLUSION: This is the first case of vesicouterine fistula repaired with a myouterine flap. This technique strengthens the repair and is especially convenient due to its easy accessibility. A myouterine flap can be utilized if the omentum is of insufficient length or absent. The risk of postoperative bowel obstruction may be decreased as compared to omental interposition. PMID- 9219128 TI - Bilateral absence of the ovaries and distal fallopian tubes. A case report. AB - BACKGROUND: Bilateral tuboovarian absence is extremely rare and is associated with infantile sexual development, primary amenorrhea and primary infertility. CASE: A 23-year-old woman presented for evaluation of primary amenorrhea. Her examination revealed hypoplastic breasts, genitalia and uterus; ovaries could not be identified. Marked estrogen deficiency was confirmed by endocrinologic testing. The karyotype was normal female. The patient was started on combined hormone replacement therapy and subsequently developed normal menses; physical maturation progressed normally. At the age of 29 she underwent diagnostic laparoscopy for evaluation of her fertility potential, at which time the absence of both ovaries and distal fallopian tubes was confirmed. CONCLUSION: Bilateral tuboovarian absence is an extremely rare cause of primary amenorrhea and is associated with infantile sexual development and primary infertility. Its etiology includes tuboovarian torsion and congenital malformation. In this case, congenital malformation appears to have been the more likely cause. PMID- 9219129 TI - Felodipine use in pregnancy. Report of three cases. AB - BACKGROUND: Calcium channel blockers are effective agents for the management of chronic hypertension and are being used with increasing frequency. If their safety and efficacy during pregnancy can be documented, women can be counseled to continue their antihypertensive agent during pregnancy. To our knowledge, the use of felodipine, a calcium channel blocker of the dihydropyridine group, during pregnancy has not been described. CASES: We report three cases of felodipine use in pregnancy by women with chronic hypertension. CONCLUSION: In women with severe hypertension (diastolic blood pressure > 100 mm Hg) who require pharmacologic treatment of it during pregnancy, felodipine appears to be an acceptable option. PMID- 9219130 TI - The low incidence of suprascapular nerve injury after primary repair of massive rotator cuff tears. AB - We measured the incidence of cuff retear and injury to the suprascapular nerve after mobilization and repair of a massive rotator cuff tear. Of one hundred four rotator cuff repairs performed over a 5-year period, 10 patients (7 men and 3 women, age range 22 to 68 years) had primary repairs of massive rotator cuff tears requiring cuff mobilization and an acromioplasty as their only procedure. These patients were evaluated at a mean of 2.5 years (range 2.0 to 3.0 years) after surgery. At follow-up electromyographic examination confirmed that 1 of the 10 patients had an iatrogenic suprascapular nerve injury, whereas ultrasound evaluation revealed that 2 of 10 repairs failed. Pain relief was achieved in the eight patients with intact repairs and not in the two with recurrent tears. All patients had some limitation of active motion or strength, especially in external rotation. Thus 7 of 10 patients had neither evidence of nerve injury nor recurrent rotator cuff tears yet still showed limited active motion or weakness. It appears that operative injury to the suprascapular nerve during cuff mobilization can occur, but other factors such as inadequate cuff muscle function are more frequently responsible for the poor functional outcomes seen after successful repairs of massive rotator cuff tears. PMID- 9219131 TI - Shoulder symptoms in healthy athletes: a comparison of outcome scoring systems. AB - We used the Rowe, ASES, UCLA, Constant-Murley, and the Simple Shoulder Test scoring systems to determine the presence and severity of shoulder symptoms in "healthy" collegiate athletes at mid-season. Intercollegiate athletes were surveyed with a single, specific, comprehensive questionnaire regarding both of their shoulders at the mid-season of their respective sport. The questionnaire compiled the previously mentioned scoring systems and additional inquiries. Shoulders were divided into three groups for analysis: dominant-never injured, nondominant-never injured, and history of injury. Significant shoulder symptoms exist in athletes during full participation in their respective sport. Pain was the most frequent symptom, with 47% of all shoulder having some degree of pain. The frequency and degree of symptoms was significantly greater in shoulders with a history of injury. The UCLA scoring system is the most sensitive for evaluating "healthy" athletes at mid-season. However, the ideal shoulder scoring system for athletes has yet to be developed. To expect a "normal" or "symptom-free" shoulder after injury or surgery may be inappropriate. This information can serve as a reference for clinicians when evaluating the results of surgery and other treatment programs. PMID- 9219132 TI - Holmium:YAG laser-assisted capsular shift in a canine model: intraarticular pressure and histologic observations. AB - The purpose of this study was to determine the initial effects of Holmium:YAG laser energy on the shoulder joint capsule. A new surgical procedure to correct shoulder joint instability uses Holmium:YAG laser energy to cause "shrinkage" of joint capsular tissues. To date there has been no information concerning an intraoperative measurable end point for the application of laser energy at surgery or the resultant depth and degree of tissue alteration. Seven greyhound dogs were used in this study. Preoperative intraarticular pressures (IAP) were measured on entry and after injection of 10 ml of solution. Laser energy was applied to the cranial medial glenohumeral ligament and joint capsule of all right shoulders with arthroscopic visualization. The unoperated left shoulders served as the control group. Six weeks after surgery pressure measurements were performed on both shoulders. A "second look" arthroscopy was performed on the shoulders. After euthanasia was performed, the anterior capsular tissues were harvested from both shoulders for histologic examination. The specimens were inspected by three blinded examiners. After 6 weeks the postoperative laser treated IAP were higher than the same joint preoperative IAP in four of six dogs for both static nondistension and 10 ml distension measurements. At this same interval the marked tissue damage of the treated capsule was easily discerned by blinded observers. On histologic evaluation the laser-treated capsule showed synovitis and pericapsular tissue reactivity. The depth of the injury was beyond the joint capsule into the pericapsular tissue. It was not possible to determine the end point of the capsular "shrinkage" operation by combined pressure/volume intraoperative measurements. There was no uniform joint capsule compliance at 6 weeks. The histologic changes were extensive in both magnitude and depth. Future studies in this animal should include decreased laser energy plus other means of monitoring the intraoperative effects of laser use. PMID- 9219133 TI - Interobserver reliability of acromial morphology classification: an anatomic study. AB - One hundred ten acromial anatomic specimens were classified by three shoulder surgeons with the classification system described by Bigliani et al. to determine the interobserver reliability. These results demonstrated a fair to poor level of interobserver reliability. Given this relatively low level of agreement, the diagnosis of impingement and rotator cuff tears should be based on clinical findings supplemented, when indicated, by rotatory cuff imaging with less diagnostic reliance placed on the assessment of acromial morphology. PMID- 9219134 TI - Closed wound drainage in shoulder surgery. AB - To evaluate the effectiveness of closed wound drainage in shoulder surgery, 300 patients were enrolled in a prospective randomized study. Three operations were studied: rotator cuff repair, anterior reconstruction for instability, and arthroplasty. One hundred patients were included in each group. All patients were evaluated for wound hematoma, infection, variation in postoperative rehabilitation caused by wound problems, and length of hospital stay. No statistical difference was found between the patients whose wounds were drained and those whose wounds were not drained. This finding existed within each category. Our data do not support the routine use of closed wound drainage in elective shoulder surgery. PMID- 9219135 TI - Anatomic variation of the coracoacromial ligament: a macroscopic and microscopic cadaveric study. AB - The operative management of the subacromial impingement syndrome includes reconstruction in cases with ruptured tendons and enlargement of the subacromial space by arthroscopic or open resection of the coracoacromial ligament and acromioplasty. In nearly 20% of all cases, however, surgical treatment fails. This study was conducted with 124 cadaver shoulders of older specimens with a balanced male/female ratio. The coracoacromial ligament did not present homogenous morphologic characteristics. We found 25.8% of all ligaments undivided, 59.7% bipartite, and 14.5% consisted of three parts. The third part, located most medially, took a hidden path to the coracoid process and was not visible during dissection until after the clavicle was resected. This medially situated third part of the coracoacromial ligament has not been described in the medical literature before the time of our investigation. It could well be responsible for persisting subacromial pain after surgery, if it is not identified and resected during surgery. PMID- 9219136 TI - Release and reattachment of the coracoacromial ligament: a cadaveric study. AB - The purpose of this study was to examine the anatomy and ability to reattach the coracoacromial ligament (CAL) after acromioplasty. Twenty-eight fresh cadaveric shoulder specimens were dissected, and the CAL dimensions and pattern of attachment were examined. The ability to reattach the CAL to the anterior acromion after acromioplasty was investigated, comparing release along the anterior acromion (traditional anterior acromioplasty) versus subperiosteal elevation from the acromial undersurface. The influence of CAL length, acromial "type," and amount of acromioplasty on reattachment were examined. In 96% of the specimens (27 of 28) confluent medial and lateral bands of the CAL insertion along the acromion precluded "selective" release. The ability to anatomically reattach the CAL was directly related to the method of ligament release (anterior release versus subperiosteal elevation) (p < 0.0001). When directly released from along the anterior acromion, the CAL could not be anatomically reattached in any specimen. Nonanatomic reattachment to a more medial anterior acromial insertion site, however, was possible in 22 (79%) of 28 specimens. Reattachment required an average medial positioning of 60% (range 16% to 88%) along the anterior acromion from the anterolateral tip. Six (21%) specimens could not be reattached despite attempted medial positioning. In contrast, when subperiosteally elevated from the anterior inferior acromion, the CAL was reattachable in all specimens, anatomically in 26 (93%) of 28. The two nonanatomic reattachments required medially reattaching the ligament's normal acromial insertion by an average of 30%. There was no relationship between specimen sex, age, presence of rotator cuff tear, type of acromion, and CAL reattachment. The confluent anatomy of the CAL does not permit "selective" release of discretely identifiable portions of the CAL (i.e., the lateral band). In cases in which CAL restoration is important, careful subperiosteal elevation from the acromial undersurface will facilitate anatomic reattachment. Standard release of the CAL from the anterior acromion precludes anatomic, and in some cases any, reattachment of the CAL. PMID- 9219137 TI - Valgus osteotomy of the humeral neck: a technique for the treatment of humerus varus. AB - Proximal humerus varus is defined by both its radiographic and clinical characteristics. Clinically significant humerus varus has a proximal humeral neck shaft angle less than 140 degrees and causes limited active abduction or forward flexion as a result of impingement of the greater tuberosity on the acromion. Weakness of the shoulder girdle is often present as well. The condition may be congenital, developmental, idiopathic, or posttraumatic in origin. Previous treatments for humerus varus have included acromionectomy and wedge osteotomy with placement of the extremity in a shoulder spica cast. This article describes a technique for treatment involving valgus osteotomy of the humeral neck and tension-band fixation. Correction of the deformity allows markedly improved function of the extremity with significant increases in active and passive abduction, forward flexion, and internal rotation. PMID- 9219138 TI - Intraarticular osteoid osteoma of the distal humerus. AB - Osteoid osteoma is a small, benign, and painful tumor most commonly affecting the extraarticular portions of the long bones, especially the femur or tibia. Osteoid osteoma of the elbow is uncommon. In this article a 19-year-old man with an osteoid osteoma of the distal humerus is presented to illustrate the diagnostic problems and unusual radiographic features of an intraarticular osteoid osteoma. PMID- 9219139 TI - "Locked" shoulder. PMID- 9219140 TI - Pectoralis major rupture with simultaneous anterior dislocation of the shoulder. PMID- 9219141 TI - Heterotopic ossification of the elbow. PMID- 9219142 TI - The costs of drug abuse consequences: a summary of research findings. AB - The purpose of this article is to provide researchers, clinicians, and policymakers with a common source of published cost estimates for drug abuse consequences. Across the broad range of potential complications associated with drug abuse, some of the cost elements are specific and directly related to drug abuse per se (e.g., drug treatment costs), while other items may be related to drug abuse (e.g., typical emergency room cost). The results presented here are based on a review of completed and ongoing studies, which include published papers, monographs, conference presentations, working papers, reports, insurance company records, newsletters, and expert judgment. Whenever possible, we report an average cost estimate and the range of available estimates. Each cost element is normalized to 1994 dollars. The results are organized by type of cost and are presented in tabular format so that the findings can be easily understood and used. PMID- 9219143 TI - Follow-up of inpatient cocaine withdrawal for cocaine-using methadone patients. AB - Significant proportions of opiate-dependent persons entering methadone treatment are also addicted to cocaine and continue to use cocaine during treatment. One standard response to cocaine use has been inpatient detoxification. This study examined the effectiveness of this procedure by comparing pre- and posttreatment urine toxicologies for methadone patients who had been hospitalized for cocaine withdrawal. The results showed a negligible effect on cocaine abstinence (less than 1 out of 10 patients abstinent 12 weeks after detox) and a modest reduction in the frequency of cocaine use (one-quarter decline in urine tests positive after 12 weeks). These findings raise serious doubts about the cost-effectiveness of inpatient cocaine detoxification. Better strategies need to be implemented to enhance the chances of remaining abstinent once detoxified. PMID- 9219144 TI - Methodological investigations for a multisite trial of auricular acupuncture for cocaine addiction: a study of active and control auricular zones. AB - We evaluated objective criteria for defining points for needle insertion prior to conducting a multisite clinical trial of auricular acupuncture for cocaine addiction. Thirty-four cocaine-abusing subjects participated in a study in which the trial's active zones (Shenmen, Liver, Lung, and Sympathetic) and control zones (located on the ear helix) were divided into quadrants and assessed along four dimensions: electrical resistance, skin discoloration, skin topography, and tenderness. Acute effects of needles inserted into points of low electrical resistance in one ear and high electrical resistance in the other were also assessed. Results showed that the active zones had lower overall electrical resistance and more subcutaneous ridges than control zones. Zones did not possess significant variability along any single dimension. Acute effects of needling high and low resistance points were similar, differing only for "fullness." Based on these findings, and in view of the difficulty of accurately measuring electrical resistance at ear points, we do not recommend the use of electrical devices for point determination in the multisite study. At present, there seems to be little scientific basis for the preselection of specific points for needle insertion within auricular zones. Needle placement should be based upon clinical judgement. PMID- 9219146 TI - Drug treatment process indicators for probationers and prediction of recidivism. AB - Research has shown that substance abuse treatment is associated with reduced criminal activity as well as reduced drug use. An increasingly important aspect of treatment evaluation, however, is understanding more about the therapeutic process involved. Based on a sample of 279 probationers assigned to residential treatment for drug use problems, the purpose of this study is to examine several elements of treatment process and how they influence recidivism. Rearrest rate during a 2-year follow-up study period was 36%, and survival analysis showed it was directly related to poorer during-treatment ratings by probationers of self esteem, counselor competence, and peer support from others in the treatment program. These were better predictor measures than background and demographic characteristics generally used in other studies, suggesting the role of therapeutic engagement in the recovery process. PMID- 9219145 TI - Sexual HIV risk among gay and bisexual male methamphetamine abusers. AB - The current report examined HIV-related high risk sexual behaviors among a small sample of gay and bisexual male methamphetamine abusers in Los Angeles. Participants were 16 methamphetamine-abusing or -dependent gay or bisexual males who participated in a treatment demonstration project between 1989 and 1993. All participants completed the NIDA/WAVE survey, a detailed inventory of HIV-related risk behaviors. Findings indicate a strong connection between methamphetamine abuse and high-risk sexual behavior. For the 12 months prior to treatment 62.5% of participants reported having anal insertive sex without a condom, and 56.3% reported having sex with someone who had HIV. Drug use before or during sex, measured on a 5-point Likert scale, was frequent (M = 4.27, SD = 0.7). Implications for treatment of gay and bisexual male methamphetamine abusers and prevention of HIV among this population are discussed. PMID- 9219147 TI - A controlled trial of imipramine for the treatment of methamphetamine dependence. AB - At the Drug Detoxification Program of the Haight Ashbury Free Clinics, we conducted a randomized clinical trial of imipramine in the treatment of methamphetamine dependence. The purposes of the trial were to test the efficacy of imipramine as a treatment for methamphetamine dependence and to establish the feasibility of conducting a controlled clinical trial at the Clinic. Thirty-two subjects were randomly assigned to receive either 10 or 150 mg/day of imiprine for 180 days. Imipramine 10 mg/day was the control. Subjects received intensive counseling. Retention in treatment was significantly longer for subjects who were treated with 150 mg of imipramine compared to control (median days: 33.0 vs. 10.5). There were no consistent differences in percent of urine samples positive for methamphetamine, Beck Depression Inventory scores, or craving. Determination of the full extent of imipramine's utility in the treatment of methamphetamine dependence awaits a larger trial. PMID- 9219148 TI - Recreational drug use concurrent with abuse or dependence on other psychoactive substances. AB - In an effort to identify instances of the non-problematic use of a drug concurrent with the problematic use of one or more other drugs, we used structured interviews to obtain comprehensive drug use histories from 48 clients admitted to an intensive outpatient program. We classified clients on the basis of whether they demonstrated evidence of concurrent problematic and non problematic drug use (Index and Probable Index cases) or only problematic drug use patterns (Non-Index cases). Both Index and Non-Index drug use patterns were about equally common in our sample. Both Index and Probable Index cases used a variety of drugs in a non-problematic manner and were generally congruent in their self-labelling of their drug use relative to their DSM-IV status for each drug used. We discuss several limitations of the study, including our reliance on retrospective, self-report data; potential problems with generalization to other populations; and possible changes in drug use patterns over time. PMID- 9219149 TI - Sexual partners of substance abuse clients: strategies for HIV/AIDS prevention. PMID- 9219150 TI - Attachment transition, addiction and therapeutic bonding--an integrative approach. AB - In the light of the attachment theory and research outlines the authors develop the concept that addiction is a delayed maladaptive attachment transition in young adults. They look into the relevance of this theoretical framework for therapy in adulthood, especially addiction therapy, connect it with an approach, that understands addiction as fear of intimacy and discuss the clinical implications of therapeutic bonding as a specialized approach for this condition. PMID- 9219151 TI - Prescribing drug of choice to illicit heroin users: the experience of a U.K. community drug team. AB - Functioning across several life domains, in the first cohort of illicit heroin users to be prescribed injectable diamorphine (pharmaceutical heroin) as an adjunct to treatment within a community drugs service, was assessed in a cross sectional study with a 6-month follow-up. Case-control matching procedures were employed to compare outcomes in this group with an oral methadone-prescribed sample, attending different clinics within the same community service and geographical locale. The Heroin Prescribed (HP) group manifested lower levels of psychopathology and showed greater retention in treatment. Although reduced, illicit heroin misuse was not eliminated; the use of other illicit substances was comparable between groups but significantly more of the HP group were using illicit cocaine. Although no differences in current physical health were apparent, the sharing of used injecting equipment was reported only in the MP group. Criminal activity appeared significantly reduced, but not eliminated, in the HP group. Implications for prescribing practice are discussed. PMID- 9219152 TI - A modified therapeutic community for the dually diagnosed. Greenhouse Program at Bellevue Hospital. AB - Techniques used in therapeutic communities may be applicable to patients dually diagnosed with mental illness and a psychoactive substance use disorder (PSUD). This study was designed to evaluate the demographics, course, and outcome for 100 patients treated in one such residential program. One hundred indigent male patients admitted to a drug-free therapeutic community for the dually diagnosed were studied on admission and over the course of their treatment, and subjects were monitored throughout their stays on the basis of observed urine toxicology tests and a clinical assessment of drug or alcohol use. The mean age of the patients was 33.8 years, and the average length of stay was 121.0 days. Thirty three of the patients completed the full 6-month program and moved on to another stable living environment. Only 12 patients had urine toxicologies positive for illicit drugs or alcohol while in the program. These findings support the possibility of applying the residential drug-free therapeutic community to dually diagnosed patients. PMID- 9219153 TI - Ultrastructural localization of osteopontin in the kidney: induction by lipopolysaccharide. AB - Osteopontin is a secreted phosphoprotein that is expressed in the normal kidney and induced during various pathologic conditions associated with tubulointerstitial injury. However, the exact cellular location of osteopontin in the kidney has been a matter of controversy, and little is known about the role of osteopontin in the kidney. The purpose of this study was to establish the cellular and intracellular distribution of osteopontin in the rat kidney under normal conditions and after injection of a bacterial endotoxin lipopolysaccharide (LPS). Animals received injections of LPS or vehicle at different time intervals from 4 to 20 h before sacrifice. Kidneys were preserved by in vivo perfusion with paraformaldehyde-lysine-periodate (PLP) and processed for light and electron microscope immunocytochemistry using monoclonal antibodies to rat osteopontin. By light microscopy, immunostaining was observed in the descending thin limb and the papillary surface epithelium of both control and LPS-treated animals. After injection of LPS, osteopontin immunostaining was observed throughout the distal nephron and was also present in segments of the proximal tubule, where it was distributed in a punctate pattern. Staining was already present 4 h after injection of LPS and was maximal 6 h after injection. Electron microscopy revealed that osteopontin immunoreactivity in the descending thin limb and distal tubule cells was located in the Golgi apparatus and in small cytoplasmic vesicles, whereas in the proximal tubule labeling was observed in the vacuolar lysosomal system. Western blot analysis demonstrated a band at approximately 70 kD and confirmed the increase in osteopontin expression after administration of LPS. These results demonstrate that osteopontin is constitutively expressed in cells of the descending thin limb and papillary surface epithelium and is induced throughout the distal tubule after administration of LPS. PMID- 9219154 TI - ETA receptor blockade prevents hypertension associated with exogenous endothelin 1 but not renal mass reduction in the rat. AB - The purpose of this study was to determine the effect of chronic ETA receptor blockade, using the orally active antagonist A-127722 in rats with reduced renal mass. The initial series of experiments was designed to characterize the effects of the ETA-selective antagonist A-127722 on arterial pressure and renal function when administered via drinking water over a 4-wk period. Male Sprague-Dawley rats were acclimated to metabolism cages, and baseline 24-h urine collections were obtained. A-127722 was placed in the drinking water at concentrations that delivered doses of 1 to 10 mg/kg per d. The compound had no effect on any of the variables measured, including arterial pressure, food and water intake, urine volume, and sodium and potassium excretion. In a separate group of rats, ETA receptor blockade was verified after 3 d of drinking water containing A-127722. Rats were anesthetized, a jugular vein catheter was inserted for infusions, and a femoral artery catheter was used for monitoring arterial pressure. The pressor response to intravenous injection of Big endothelin-1 (1 nmol/kg, intravenously) was inhibited by > 50% in rats given A-127722 at 10 mg/kg per d, which confirms the efficacy of A-127722 in blocking ETA-mediated responses when placed in drinking water. In an additional series of experiments, rats were anesthetized, the right kidney was removed, and two of three major branches of the left renal artery were ligated. After recovery, rats were returned to their cages and given A-127722 in the drinking water to deliver 1 or 10 mg/kg per d. Control rats underwent the same surgical procedures but were given tap water to drink. After 4 wk, rats that were treated with A-127722 developed similar increases in arterial pressure and urinary protein excretion as rats that received tap water. Therefore, although the ETA receptor antagonist A-127722 can inhibit ETA-mediated hypertension, it has no effect on hypertension produced by a reduction in renal mass. It is concluded that ETA receptor activation does not play a significant role in the functional derangements associated with renal mass reduction in the rat. PMID- 9219155 TI - Renal protection by a dual ETA/ETB endothelin antagonist, L-754,142, after aortic cross-clamping in the dog. AB - Renal insufficiency is a significant complication that occurs after surgical procedures, requiring cross-clamping of the aorta. The mechanism for this renal dysfunction is currently not known, but studies suggest a potential role of endothelin in mediating the insufficiency. Accordingly, the role of endothelin was assessed using the nonpeptidyl, dual ETA/ETB endothelin antagonist L-754,142 in a model of renal insufficiency in the anesthetized dog induced by cross clamping the suprarenal aorta for 60 min, followed by 2 h of reperfusion. In vehicle-treated animals (saline, n = 8) after 2 h of reperfusion, plasma [ET-1] increased 66% and renal blood flow (RBF) was reduced by 38% compared with baseline. This decline was associated with an 84% increase in renal vascular resistance and a 54% reduction in GFR (baseline, 46 +/- 5 ml/min; 21 +/- 3 ml/min at 2 h; P < 0.01) and sodium reabsorption (baseline, 6.7 +/- 0.7 microEq/min; 3.0 +/- 0.5 microEq/min at 2 h, P < 0.01). After baseline measurements, pretreatment with L-754,142 at 0.3 mg/kg bolus + 0.1 mg/kg per h continuous infusion (low dose; n = 8) or 3.0 mg/kg bolus + 1 mg/kg per h infusion (high dose; n = 8) initiated 45 min before aortic cross-clamp led to a dose-dependent normalization of RBF and renal vascular resistance within 2 h of cross-clamp removal. GFR was also improved and returned to within 75% of baseline (P < 0.01 versus vehicle) by 2 h of reperfusion with L-754,142 (baseline, 55 +/- 5 ml/min; 42 +/- 5 ml/min at 2 h with the high dose). The improvement of GFR with L-754,142 treatment was associated with a preservation of sodium reabsorption compared with vehicle treated animals. This study supports a role of endothelin in the pathogenesis of renal insufficiency after aortic cross-clamping and demonstrates that pretreatment with the dual ETA/ETB endothelin antagonist L-754,142 preserves RBF and sodium reabsorption, leading to a significant improvement in GFR. PMID- 9219156 TI - Angiotensinogen gene expression is stimulated by the cAMP-responsive element binding protein in opossum kidney cells. AB - It has been reported previously that the addition of isoproterenol or forskolin stimulates the expression of the angiotensinogen (ANG) gene in opossum kidney (OK) 27 cells, an OK cell line with a fusion gene containing the 5'-flanking regulatory sequence of the rat ANG gene fused with a human growth hormone (hGH) gene as a reporter, pOGH (ANG N-1498/+18), permanently integrated into their genomes. To investigate whether the effect of isoproterenol or forskolin on the expression of the ANG gene is mediated via the nuclear 43-kD cAMP-responsive element binding protein (CREB), OK 27 cells were transiently transfected with an expression plasmid containing the cDNA for the 43-kD CREB (pRSV/CREB). The level of expression of the pOGH (ANG N-1498/+18) in OK 27 cells was estimated by the amount of immunoreactive hGH secreted into the culture medium. Transfection of pRSV/CREB alone stimulated the expression of pOGH (ANG N-1498/+18). The addition of isoproterenol or forskolin further enhanced the stimulatory effect of pRSV/ CREB on the expression of pOGH (ANG N-1498/+18). The enhancing effect of isoproterenol was inhibited by the presence of propranolol (an inhibitor of beta adrenoceptors) and (R)-p-adenosine 3'5'-cyclic monophospho-orthioate (Rp)-cAMP (an inhibitor of cAMP-dependent protein kinase A I and II). Transfection of pRSV/CREB had no effect on the expression of thymidine kinase growth hormone in OK 13 cells, an OK cell line with a fusion gene containing the promoter/enhancer DNA sequence of the viral thymidine-kinase gene fused with an hGH gene as a reporter, thymidine kinase growth hormone, permanently integrated into their genomes. These studies demonstrate that isoproterenol stimulates the expression of ANG gene via the cAMP-dependent protein kinase A and probably via the interaction of the 43-kD CREB with the 5'-flanking region of the ANG gene. Our data indicate that the nuclear 43-kD CREB may have a modulatory role on the expression of the ANG gene in OK cells. PMID- 9219158 TI - Neutrophil superoxide release is required for spontaneous and FMLP-mediated but not for TNF alpha-mediated apoptosis. AB - Polymorphonuclear leukocyte (PMN) lifespan is characterized by both rapid production and apoptotic cell death. The mechanisms triggering apoptosis in PMN are not completely understood. In this study, the relationship of neutrophil activation and apoptosis as related to released superoxide was investigated. PMN apoptosis was detected by DNA fragmentation, and ultraviolet and light microscopy, and was quantified by flow cytometry; superoxide release was measured by superoxide dismutase-inhibitable reduction of ferricytochrome C. Incubation of PMN with 20 ng/ml tumor necrosis factor (TNF)alpha induced superoxide release (8.8 +/- 7.5 nmol O2-/30 min, n = 7) in normal PMN and also resulted in apoptosis within 2 h, whereas a subactivating dose of 2 ng/ml TNF alpha, which did not trigger superoxide release (3.1 +/- 1.7 nmol O2-, n = 10), did facilitate apoptosis, although to a lesser degree. PMN cultured under nonstimulating conditions underwent apoptotic cell death after 8 h. Exogenous superoxide dismutase did not inhibit apoptosis induced by 20 ng/ml TNF alpha. No upregulation of endogenous manganese superoxide dismutase mRNA expression was observed in response to TNF alpha as measured by reverse transcription PCR. Formyl-methionyl-leucyl-phenylalanine (FMLP) stimulation (10(-7) M) resulting in superoxide release of 31.7 +/- 6.1 nmol O2-/30 min (n = 10) also significantly increased the percentage of apoptosis, but at 24 h (P < 0.05). Exogenous superoxide dismutase did inhibit FMLP-induced apoptosis, as well as apoptosis due to aging in culture. In conclusion, aging and FMLP-stimulated PMN undergo apoptosis by a superoxide release-dependent pathway, whereas TNF alpha facilitated apoptosis appears to be unrelated to respiratory burst oxidase activity. PMID- 9219157 TI - Calcium-regulated protein tyrosine phosphorylation is required for endothelin-1 to induce prostaglandin endoperoxide synthase-2 mRNA expression and protein synthesis in mesangial cells. AB - The role of endothelin (ET)-1-mediated cytosolic calcium ([Ca2+]i) elevation in regulating ET-1-induced prostaglandin endoperoxide synthase, prostaglandin G/H synthase (PGHS)-2 mRNA expression and protein synthesis was investigated in mesangial cells (MC). Ionomycin, a calcium ionophore, and thapsigargin, an inhibitor of calcium ATPase, mimicked the ET-1-stimulated PGHS-2 mRNA and protein induction. Inhibition of [Ca2+]i increases with (2-?C2-bis-(carboxymethyl)-amino 5 methylphenoxy]methyl?-6-methoxy-8-bis-(carboxymethyl)-aminoquinoline tetra (acetoxymethyl)ester (Quin/AM), a calcium chelator, or with the combined presence of [8-(diethylamino)-octyl-3,4,5-trimethoxybenzoate, HCl] (TMB), an inhibitor of intracellular calcium stores release, and ethyleneglycol-bis-(beta-aminoethyl)- N,N,N',N'-tetra-acetic acid (EGTA) suppressed ET-1, as well as ionomycin and thapsigargin-mediated PGHS-2 mRNA and protein formation. Also, the ET-1-, ionomycin-, and thapsigargin-induced PGHS-2 mRNA expression and protein formation was inhibited in MC pretreated with inhibitors of calcium calmodulin kinase. In contrast, these conditions did not inhibit interleukin (IL)-1-induced PGHS-2 mRNA expression and protein synthesis. Pretreatment with tyrosine kinase inhibitors abolished the ET-1-, ionomycin-, thapsigargin-, and IL-1-mediated PGHS-2 mRNA and protein induction. ET-1-, ionomycin-, and thapsigargin- induced protein tyrosine phosphorylation, but not IL-1-induced protein tyrosine phosphorylation, was suppressed by inhibiting either [Ca2+]i elevation or calcium calmodulin kinase activation. It was concluded that elevation of [Ca2+]i and activation of calcium calmodulin kinases are upstream mediators of ET-1-induced PGHS-2 gene expression through activation of non-receptor-linked protein tyrosine kinase in MC. PMID- 9219159 TI - Th2 responses induce humorally mediated injury in experimental anti-glomerular basement membrane glomerulonephritis. AB - Acute autologous phase anti-glomerular basement membrane glomerulonephritis was compared in Th1-prone (C57BL/6) and Th2-prone (BALB/c) mice. Sensitized BALB/c mice, given a subnephritogenic intravenous dose of anti-mouse glomerular basement membrane globulin, developed acute glomerulonephritis characterized by marked proteinuria and glomerular deposition of mouse immunoglobulin and complement. A significant glomerular neutrophil influx was observed, but few T cells and macrophages were present. C57BL/6 mice, given the same dose of disease-inducing globulin, also developed acute glomerulonephritis, although their proteinuria was significantly less. Glomerular deposition of mouse immunoglobulin and complement and the influx of neutrophils were also significantly less than in BALB/c mice. However, their glomerular accumulation of macrophages and T cells was significantly greater. Complement depletion attenuated neutrophil influx and proteinuria in BALB/c mice but did not affect T cell or macrophage accumulation or proteinuria in C57BL/6 mice. CD4+ T cell depletion significantly reduced glomerular macrophage, T cell influx, and proteinuria in C57BL/6 mice, but had no effect on proteinuria or neutrophil influx in BALB/c mice. Thus, immune responses to planted glomerular antigens in Th2-prone mice induce acute injury as a result of antibody deposition, complement activation, and neutrophil influx, whereas immune responses to the same antigen in Th1-prone mice induce delayed-type hypersensitivity-like lesions in affected glomeruli. PMID- 9219160 TI - Preservation of intercalated cell H(+)-ATPase in two patients with lupus nephritis and hyperkalemic distal renal tubular acidosis. AB - In patients with Sjogren's syndrome and a secretory-defect distal renal tubular acidosis (dRTA), absence of vacuolar H(+)-ATPase from collecting duct intercalated cells has been reported. The H(+)-ATPase was examined in two patients with lupus nephritis and hyperkalemic (presumed voltage defect) dRTA. Both patients had a positive urine anion gap, alkaline urine despite acidemia, no rise in urine PCO2 with alkaluria, a urine pH > 5.5, and urine potassium excretion rate not significantly increased after 80 mg of intravenous furosemide. In both patients, immunocytochemistry of renal biopsy frozen sections with an anti-H(+)-ATPase monoclonal antibody showed bright staining of the proximal tubule brush border and collecting duct intercalated cells. In one patient, routine immunofluorescence analysis of a frozen section of her kidney biopsy with antihuman IgG showed staining of the collecting duct, indicative of autoantibodies to this segment. Moreover, rat kidney sections incubated with her serum showed labeling of the intercalated cells. On immunoblots of human kidney microsomal membranes performed with serum from both patients, an immunoreactive polypeptide was observed at M(r) approximately 56 kD that was not seen with control serum. Neither patient's sera reacted with affinity-purified bovine H(+) ATPase or with lysates from 293 cell fibroblasts in which either of both isoforms of the human H(+)-ATPase B subunit (56 kD) were expressed. These findings demonstrate that the spectrum of dRTA includes the preservation of H(+)-ATPase in intercalated cells, in patients with presumed voltage defect dRTA. Moreover, some patients may have autoantibodies to the intercalated cells that are not directed to subunits of the H(+)-ATPase. PMID- 9219161 TI - Mutation in mitochondrial tRNA(Leu(UUR)) gene associated with progressive kidney disease. AB - Several studies show an association of a guanine for adenine substitution (A-->G) at position 3243 in mitochondrial DNA (mtDNA) with a recently recognized diabetic subtype designated maternally inherited diabetes and deafness (MIDD). This mutation shows heterogeneity in its phenotypic expression as is apparent from its association with several other syndromes. Screening for the 3243A-->G mutation in mtDNA was performed in those diabetic patients attending the Leiden University Hospital diabetics clinic who had a history of maternally inherited diabetes, sensorineural hearing loss, or both. Four individuals from three unrelated families were identified who suffered from progressive nondiabetic kidney disease in association with diabetes mellitus and hearing loss. The mode of inheritance suggested maternal transmission. The combination of renal failure and hearing loss had been misdiagnosed as Alport syndrome in three of the four individuals. Therefore, in addition to these three families, another 63 unrelated patients with possible Alport syndrome were selected at random. DNA from peripheral blood and other tissues from members of the three families and from the 63 additional Alport syndrome patients was examined for the presence of the 3243A-->G mutation in mtDNA. The mutation was detected in heteroplasmic form in the four patients and their maternal relatives. Also, one of the 63 suspected Alport syndrome patients showed heteroplasmy for the 3243 mutation. These data show the existence of a kidney disease that is characterized by the presence of the A-->G mutation at position 3243 in the mtDNA. PMID- 9219162 TI - Prevention of radiocontrast-induced nephropathy by adenosine antagonists in rats with chronic nitric oxide deficiency. AB - To evaluate therapeutic options for the prevention of radiocontrast media (RCM) induced nephropathy, a model was developed in which rats received NG-nitro-L arginine methyl ester (L-NAME) for 10 wk in order to inhibit nitric oxide (NO) synthetase. This study tests the hypothesis that infusion of an adenosine antagonist before RCM application may avoid the vasoconstrictive response in NO depleted rats. Rats received L-NAME for 10 wk orally (50 mg/L drinking water) to achieve NO depletion. Renal function was determined by [3H]inulin clearance for analysis of the GFR and by flowmetry for assessing renal blood flow (RBF). After a control clearance period (baseline clearance period), the renal response to RCM application (sodium diatrizoate, 2 ml/kg body wt) was measured two times every 30 min starting 30 min after RCM application (clearance periods 1 and 2). L-NAME rats and control rats received two adenosine antagonists. The nonselective adenosine antagonist theophylline was given as an initial bolus of 50 mumol/kg body wt within 10 min, followed by continuous infusion of 100 mumol/kg body wt per h, and the specific adenosine A1-receptor antagonist 8-cyclopentyl-1,3 dipropylxanthine (DPCPX) was given as a bolus of 100 micrograms/kg body wt before RCM application. Results were compared with vehicle infusion. In the control group, no significant change of GFR or RBF could be detected after application of RCM with or without prior infusion of DPCPX or theophylline. In L-NAME rats, RBF decreased significantly after RCM application (baseline, 5.6 +/- 0.2 ml/min; first clearance period, 4.6 +/- 0.3 ml/min [P < 0.05]; second clearance period, 4.3 +/- 0.3 [P < 0.01]). GFR was also reduced in L-NAME rats without previous infusion of theophylline or DPCPX (baseline, 0.95 +/- 0.1 ml/min; first clearance period, 0.83 +/- 0.1 ml/min; second clearance period, 0.69 +/- 0.1 ml/min [P = 0.058]). Prior treatment with either theophylline or DPCPX resulted in complete protection against a decline of RBF and GFR induced by RCM in L-NAME rats. Rats with chronic NO blockade showed a significant increase of the renal vasoconstrictive effect of contrast media. Application of L-NAME in rats seems to constitute a suitable animal model to study the pathophysiology of radiocontrast media-induced nephropathy. In this animal model, administration of adenosine antagonists prevented the decline of GFR and RBF. PMID- 9219163 TI - Enhanced collagen synthesis in cultured skin fibroblasts from insulin-dependent diabetic patients with nephropathy. AB - Excessive production and deposition of extracellular matrix proteins are characteristic features of diabetic nephropathy. This study tests the hypothesis that cells from diabetic patients who develop nephropathy have a disturbance in collagen metabolism compared with cells from diabetic patients without complications. Kinetics of overall collagen metabolism and total protein synthesis were examined in serially passaged, subconfluent, quiescent skin fibroblasts cultured in either normal (5 mM) or high (25 mM) glucose concentrations from 14 insulin-dependent diabetic (IDDM) patients with nephropathy; 14 IDDM patients without nephropathy matched for age, diabetes duration, and body mass index; and 14 healthy subjects. Fibroblasts were incubated in the presence of 2 microCi/ml [3H]proline, and after labeling the incorporation of [3H]proline into total protein, collagen (collagenase-sensitive material), and noncollagen proteins (collagenase-resistant material) was determined at different time points. Collagen degradation was determined in pulse chase experiments by following the residual collagen-bound radioactivity after incubation for 8 h with 10 microCi/ml [3H]proline. In high glucose concentrations (25 mM), overall collagen synthesis (measured as [3H]proline incorporation into extracellular and intracellular collagenase-sensitive material) was significantly greater in the patients with nephropathy (mean +/- SEM after a 24-h labeling period: 7189 +/- 671 dpm/10(6) cells) than in the patients without (4341 +/- 267 dpm/10(6) cells; P < 0.01) or healthy control subjects (3836 +/- 234 dpm/10(6) cells; P < 0.01). No significant differences were observed in noncollagen protein production or in collagen degradation rates among the three groups of subjects. In the presence of normal glucose concentrations (5 mM), collagen synthesis was lower in all groups studied, but the differences between IDDM patients with nephropathy and those without remained unaltered. These results suggest that long term cultured fibroblasts derived from diabetic patients with nephropathy exhibit an abnormality in collagen metabolism. Cells from long-standing diabetic patients without nephropathy have normal collagen metabolism. The increased collagen synthesis is likely to be intrinsic to those diabetic patients susceptible to nephropathy and may play an important role in the sclerotic processes that occur in the kidneys, arteries, and heart. PMID- 9219164 TI - Effect of angiotensin-converting enzyme inhibition in HIV-associated nephropathy. AB - Angiotensin-converting enzyme inhibition (ACEI) delays progression of diabetic and nondiabetic renal disease. This study examined the effect of fosinopril, 10 mg by mouth daily, in HIV-associated nephropathy (HIV-AN). Twenty patients with HIV-AN were studied. Of 11 patients with non-nephrotic-range proteinuria, 7 received treatment and 4 did not. Average baseline creatinine (mg/dl) for treated and nontreated patients was 1.3 +/- 0.24 and 1.0 +/- 0.25, respectively (P = 0.07). At 24 wk, creatinine of treated and nontreated patients was 1.5 +/- 0.34 and 4.9 +/- 2.4 (P = 0.006). Average baseline 24-h urine protein excretion (g/d) for treated and nontreated patients was 1.6 +/- 0.68 and 0.78 +/- 0.39, respectively (P = 0.02). At 24 wk, 24-h protein excretion of treated and non treated patients was 1.25 +/- 0.86 and 8.5 +/- 1.4 (P = 0.006). Of nine patients with nephrotic-range proteinuria, five were treated and four were not. Average baseline creatinine for treated and nontreated patients was 1.7 +/- 0.46 and 1.9 +/- 0.42, respectively (P = 0.4). At 12 wk, creatinine for treated and nontreated patients was 2.0 +/- 1.0 and 9.2 +/- 2.0 (P = 0.02). The baseline 24-h protein excretion for treated and nontreated patients was 5.4 +/- 1.6 and 5.2 +/- 0.97 (P = 0.9). At 12 wk, 24-h protein excretion for treated and nontreated was 2.8 +/- 1.0 and 10.5 +/- 3.5 (P = 0.008). These preliminary data suggest that treatment with ACEI may stabilize serum creatinine and 24-h protein excretion for up to 24 wk in patients with non-nephrotic-range proteinuria and for up to 12 wk in patients with nephrotic-range proteinuria when initial serum creatinine is < or = 2.0 mg/dl. Furthermore, the renin-angiotensin system may play a role in HIV-AN, and early treatment with ACEI may be beneficial in HIV-AN. PMID- 9219165 TI - Long-term effects of erythropoietin therapy on fistula stenosis and plasma concentrations of PDGF and MCP-1 in hemodialysis patients. AB - Among the adverse effects possibly associated with the use of erythropoietin (EPO) in hemodialysis patients is an increased incidence of thrombosis of the vascular access. However, little is known about the effect of EPO on the stenotic lesion in the venous outflow system, which is the leading cause of fistula thrombosis. This study was designed to explore the long-term effects of EPO treatment on progressive fistula stenosis and the plasma concentrations of some potential mediators of neointimal hyperplasia. A cross-sectional and 3-yr prospective, placebo-controlled, pilot study was performed in 30 hemodialysis patients with native arteriovenous fistula. Sixteen patients received EPO and 14 received a placebo. Venous dialysis pressure, urea recirculation, color Doppler sonography, and angiography were used to monitor vascular access patency. Compared with 60 healthy subjects, the hemodialysis patients had elevated plasma levels of platelet-derived growth factor, monocyte chemoattractant protein-1, and interleukin 6, three proteins that might be involved in the neointima formation regulating the proliferation of vascular smooth muscle cells. In addition, these patients had numerous endothelial and hemostatic abnormalities that indicated a thrombophilic state. Eleven patients, six (37.5%) receiving EPO and five (35.7%) taking placebo, developed a progressive stenosis in the venous circuit of the fistula. There was no significant difference in the vascular access, event-free survival over 36 mo between patients receiving EPO therapy and placebo. EPO induced a significant decrease in the plasma values of platelet-derived growth factor and vascular cell adhesion molecule-1 and an increase of monocyte chemoattractant protein-1 concentration. After EPO withdrawal, these parameters returned to pretreatment levels. In conclusion, long-term EPO therapy does not increase the risk of progressive stenosis of native arteriovenous fistula. The use of erythropoietin does not induce any prothrombotic change in hemostatic parameters, and further studies are required to elucidate the theoretically beneficial effects on the plasma concentration of some potential mediators of neointimal formation. PMID- 9219166 TI - Favorable effect of hemodialysis on decreased serum antioxidant activity in hemodialysis patients demonstrated by electron spin resonance. AB - There is considerable evidence that uremic patients are in a highly peroxidative state. The purpose of this study was to investigate the serum antioxidant activity that may regulate, or represent, the redox state in vivo. Serum from pre and posthemodialysis patients and from healthy control subjects was added to a system generating the hydroxyl radical, and then the signal intensities of reactive oxygen species were measured by electron spin resonance and spin trapping technique. The electron spin resonance signals of the reaction mixture containing prehemodialysis sera were significantly stronger than those of the reaction mixture containing healthy sera (P < 0.001, n = 19), and there was no significant difference in the signals between the reaction mixture containing posthemodialysis and healthy sera. These findings demonstrated that serum antioxidant activity in hemodialysis patients is significantly decreased, and this pathological condition is improved by hemodialysis treatment. PMID- 9219167 TI - Impact of pretransplantation GB virus C infection on the outcome of renal transplantation. AB - Among renal transplant recipients with posttransplantation liver disease, the etiology remains unknown in 10 to 16% of patients. The discovery of yet another parenterally transmitted hepatitis virus, GB virus C (GBV-C), has opened avenues to study the prevalence and risk factors for GBV-C infection among patients undergoing renal transplantation and its impact on posttransplantation clinical outcomes. A cohort of 103 randomly selected recipients of kidneys were examined from anti-hepatitis C virus (HCV)-negative donors between 1986 and 1990. Pretransplantation sera were available in 99 of 103 (96%) recipients and were tested for anti-HCV, using a second-generation ELISA, and for GBV-C RNA by reverse transcription PCR. Pretransplantation GBV-C RNA was present in 18 of 99 (18%, 95% confidence interval [CI], 17.2 to 18.8%) recipients. GBV-C RNA was present in 5 of 22 (23%) anti-HCV-positive recipients compared with 13 of 77 (17%) anti-HCV-negative recipients (P = 0.53). The median number of pretransplantation blood transfusion among recipients with GBV-C RNA before transplantation was significantly higher than among recipients without GBV-C RNA (10 versus 7, P = 0.05). Posttransplantation liver disease and non-A, non-B hepatitis (NANBH) was observed in 35 and 18%, respectively, of GBV-C RNA-positive recipients compared with 28 and 10%, respectively, of GBV-C RNA-negative recipients. Using Cox regression analysis, the relative risk (RR) of posttransplantation liver disease among recipients with GBV-C RNA before transplantation was 1.37 (95% CI, 0.55 to 3.41), and posttransplantation NANBH was 2.09 (95% CI, 0.64 to 6.79). The RR of graft loss and death were not increased (0.88 and 0.92, respectively). When adjusted for pretransplantation anti-HCV, the RR of posttransplantation liver disease, NANBH, graft loss, and death did not change appreciably. In summary, although a higher risk of posttransplantation liver disease was observed among recipients with pretransplantation GBV-C infection, the analyses presented here do not allow for a precise estimate of this risk. PMID- 9219168 TI - Suppression of anti-glomerular basement membrane nephritis by administration of anti-monocyte chemoattractant protein-1 antibody in WKY rats. AB - Anti-glomerular basement membrane (GBM) nephritis in WKY rats is characterized by an accumulation of CD8-positive lymphocytes (CD8+ lym) and monocytes/macrophages (Mo/M psi) in the glomeruli and crescent formation. In the study presented here, the involvement of a chemokine for Mo/M psi, monocyte chemoattractant protein-1 (MCP-1), was examined in this model. An intense induction of mRNA for MCP-1 in the glomeruli and a suppressive effect of anti-MCP-1 antibody administration on the glomerular Mo/M psi accumulation and proteinuria were found. MCP-1 mRNA was expressed intensely in the glomeruli 4 d after the anti-GBM antibody injection. When MCP-1 was neutralized with anti-MCP-1 antibody administration, the number of Mo/M psi infiltrating in the glomeruli decreased by 34.7% (19.6 +/- 7.1 versus 30.0 +/- 6.0 per glomerular cross-section, P < 0.01) and proteinuria by 66.2% (15.6 +/- 9.3 versus 46.1 +/- 15.1 mg/d, P < 0.01) at day 4. In contrast, the number of CD8+ lym accumulating in the glomeruli was not affected significantly (6.6 +/- 2.7 versus 6.0 +/- 3.0 per glomerular cross-section, P > 0.05). However, the treatment with the anti-MCP-1 antibody did not reduce Mo/M psi infiltration, urinary protein excretion, and crescent formation at day 8. These data suggest that MCP-1 plays a role in the glomerular accumulation of Mo/M psi, and that the infiltrating Mo/M psi cause glomerular injury and increased excretion of protein in the urine. PMID- 9219169 TI - Hypokalemia--consequences, causes, and correction. PMID- 9219170 TI - The natural history of the renal manifestations of systemic lupus erythematosus. 1964. PMID- 9219171 TI - Mesangial lupus nephritis with associated nephrotic syndrome. AB - Patients with mesangial proliferative lupus glomerulonephritis (World Health Organization class II) are generally believed to have only mild to moderate proteinuria and normal renal function. However, there have been several reports of patients with mesangial lupus with nephrotic-range proteinuria. In this report, we present two additional cases and review the literature. Of seven reported cases, persistent nephrotic syndrome was observed in four, morphologic transformation occurred in three, and all but one presented with varying degrees of azotemia. These cases reinforce the concept that in systemic lupus erythematosus, laboratory findings may not correlate well with the underlying glomerular lesion, and therefore, the renal biopsy is an essential clinical tool in the approach to lupus nephritis. PMID- 9219172 TI - Propylthiouracil-induced diffuse proliferative lupus nephritis: review of immunological complications. AB - Propylthiouracil (PTU), used to treat Graves' disease, occasionally induces a lupus-like syndrome. A 39-year-old woman developed clinical manifestations of systemic lupus erythematosus with rash, serositis, myocarditis, and acute renal insufficiency, associated with serologies for lupus, after 3 wk of exposure to the drug. Renal biopsy revealed diffuse proliferative lupus nephritis. This article reviews the side effects of PTU and the literature on PTU-induced nephrotoxicity. Possible mechanisms and management of drug-induced lupus nephritis are also reviewed. To facilitate early and specific intervention, clinicians should be aware of the propensity of PTU to cause lupus-like syndromes with renal involvement. PMID- 9219174 TI - Water household of the common carp, Cyprinus carpio, when submitted to an osmotic challenge, as determined by diffusion-weighted magnetic resonance imaging at 7 T. AB - In vivo diffusion-weighted magnetic resonance imaging (MRI) was used to determine the effects of an osmotic challenge (1% NaCl) to a freshwater fish, the common carp (Cyprinus carpio). The imaged region covered organs such as the swimbladder, the liver, the kidney, the intestine, the spinal cord, and muscle tissue. A striking difference between salt-treated and control fish was found in the liver. The apparent diffusion coefficient value of livers from control fish was (0.39 +/ 0.16) 10(-9) m2/s and of salt-treated fish was (1.23 +/- 0.14) 10(-9) m2/s, which points to an increase in extracellular water content. These results were partially confirmed by a decrease in dry/wet weight ratio of the liver tissue. We also found increased levels of stress proteins in liver tissue. The Q factor of the applied radiofrequency coil dropped dramatically when we performed experiments with salt-exposed fish, indicating an increased conductivity resulting from the increased ion concentration and osmolarity of the fish. The data on plasma osmolarity of salt-exposed fish confirm a significant osmolarity increase upon salt exposure (from 334 to 430 mOsm/kg) and exceeded the osmolarity of the salt water (324 mOsm/kg), indicating that carp tend to cope with an increased salinity by increasing the internal osmolarity (hyperosmotic regulation). These data demonstrate that diffusion-weighted MRI might be a useful and noninvasive tool in the study of osmotic challenges of aquatic organisms. PMID- 9219173 TI - Young Investigator Award presentation at the 13th annual meeting of the ESMRMB, September 1996, Prague. Quantification of pulmonary water compartments by magnetic resonance. AB - The purpose of this study was to quantify pulmonary water compartments of total, intravascular, and extravascular lung water in excised and perfused sheep lungs with the use of magnetic resonance imaging techniques. Total lung water was measured by proton density maps calculated from multi-spin-echo images. Intravascular lung water was evaluated by magnetic resonance angiography before and after injection of gadolinium diethylenetriamine penta-acetic acid polylysine, a macromolecular paramagnetic contrast agent. Intravascular lung water was calculated from signal intensity histogram changes comparing pre- and postcontrast angiograms. Extravascular water was calculated as the difference between total and intravascular lung water. Quantities of total and intravascular lung water measured by magnetic resonance techniques were compared to reference results obtained from wet/dry weight gravimetry and Evans blue dilution performed after imaging. Magnetic resonance and reference results correlated significantly (total lung water: r = 0.93, p < 0.001; intravascular lung water: r = 0.80, p < 0.001; extravascular lung water: r = 0.89, p < 0.001). Therefore, we conclude that quantitative magnetic resonance techniques are potentially useful for the clinical evaluation of pulmonary water compartments. PMID- 9219175 TI - High resolution magnetic resonance imaging application in anatomy: the extensor digitorum muscle insertion on the first phalanx. AB - Classical dissection may give unsatisfactory results because of the presence of artifacts due to both the embalming process and displacement of the anatomical structures. This spatial disturbance could explain the divergent descriptions found in the literature about the presence, or the absence, of an insertion of the extensor digitorum muscle (ED) at the first phalanx (P1). Preliminary experiments by Van Sint Jan et al. (1996) found the same contradiction: dissections did not show a real tendon attachment, whereas a functional experiment seemed to show that "something" should exist between ED and P1 to explain the results. This paper presents the results of an in vitro MRI study of this anatomical area. A 7-T NMR microscope was used to collect accurate, noninvasive data. Subsequently, surface rendering was performed to visualize the structures in a three-dimensional manner. The results of this MRI study, together with functional data obtained in an earlier study, showed that no real insertion of ED on P1 exists. However, some collagenic fibers were occasionally-observed running from the ventral aspect of ED to both P1 and the metacarpo-phalangeal joint capsule. Those few collagenic fibers would play a secondary role in the extension of P1. PMID- 9219176 TI - Is there a correlation in breast carcinomas between tumor size and number of tumor vessels detected by gadolinium-enhanced magnetic resonance mammography? AB - Tumor vessels are known as a sign of malignancy in breast tumors. Is there a correlation between tumor size and the number of vessels in cases of breast tumor examined by dynamic gadolinium (Gd)-enhanced MR imaging? Eighteen patients (mean age, 46 +/- 7 years) underwent dynamic Gd-enhanced MR imaging of the breast by three-dimensional gradient echo sequence using thin-layer technique (2.5 mm) at 1.5T. The dynamic study included one precontrast and four postcontrast sequences (every 90 seconds) in coronal slices. Postprocessing by subtraction method and reconstruction in both transverse and sagittal planes were performed. All carcinomas showed rapid Gd enhancement. Tumor size (0.5 to 31.5 cm3; mean, 6.3 +/ 3.7 cm3) and number of vessels (1 to 10; mean, 3 +/- 2.1) were detected in summation of all three directions. A significant correlation was found between number of vessels and tumor size (r = 0.787, p < or = 0.01). Breast tumor size significantly correlated with the number of vessels detected by Gd-enhanced MR mammography. The introduced method is a further important step in differentiating a carcinoma from a benign lesion. PMID- 9219177 TI - Low-frequency quadrature mode birdcage resonator. AB - The birdcage resonator is frequently used in conventional MRI because of its excellent attributes. Its use in low-field MRI is restricted to field strengths higher than, for example, 0.1 T, dependent on the size of the coil. This is because of the intrinsically low inductance value of the birdcage coils. Furthermore, the sensitivity of the birdcage at low field strengths is significantly lower when compared to, for example, the solenoid. Both problems can be overcome with the multiturn technique and a novel wound birdcage coil. The quadrature mode wound birdcage coil presented in this paper can be used at frequencies as low as 100 kHz. Its sensitivity is also increased when compared to the conventional strip-ring birdcage. Homogeneity, effective volume, and methods to increase the resonator bandwidth to match the signal bandwidth are left intact. The latter is a typical low-field problem. PMID- 9219178 TI - Magnetic resonance susceptibility contrast induced by capillaries: a numerical comparison of two models. AB - Monte-Carlo computer simulations have proven to be very powerful tools for the analysis of the magnetization decay induced by susceptibility gradients, as well for contrast agent characterization, as for the BOLD effect allowing fMRI. A recent vasculature model containing capillaries and venules uses homogeneous magnetized cylinders as models for the vessels. This modeling is questioned by comparing results obtained from simulation results based on two different models, one using homogeneous cylinders and another taking into account the existence of red blood cells, treated as homogeneous magnetized spheres. The results show the nonequivalence of both models, with the modeling by cylinders systematically overestimating the transverse relaxation rates, and the difference increasing with the adopted value of the diffusion coefficient. The discrepancy is attributed to the dominating role, regarding relaxation, of the local magnetic field in the immediate vicinity of the capillaries, which results in the suggestion of elaborating a "mixed modeling": the analytical expressions corresponding to the homogeneous cylinder model could be used except when the spin packets are wandering in the immediate vicinity of the capillaries, where accounting for the existence of individual red blood cells (whose motion may be neglected) seems unavoidable. PMID- 9219180 TI - Pulsed gradient analysis using a dedicated magnetometer. AB - In magnetic resonance imaging and spectroscopy, knowledge of the magnetic field gradient behavior is very important. This work describes a simple way to characterize the temporal and spatial dependence of the main magnetic field when a gradient is switched. Records are performed with a home-built magnetometer. This device is controlled by a personal computer for recording and processing the NMR signals from an array of small probes spatially distributed and switched by the magnetometer. We present results of measurements on a 2-T superconducting magnet. These results show the residual defects of an active shielded gradient coils system. PMID- 9219179 TI - Use of spin-traps during warm ischemia-reperfusion in rat liver: comparative effect on energetic metabolism studied using 31P nuclear magnetic resonance. AB - Detection of free radicals by electron spin resonance (ESR) proves the involvement of reactive oxygen species (ROS) in reperfused organ injuries. Spin traps are known to ameliorate hemodynamic parameters in an isolated postischemic heart. The effects of 5 mmol/L DMPO (5,5-dimethyl-1-pyrroline-N-oxide) or DEPMPO (5-(diethlphosphoryl)-5-methyl-1-pyrroline N-oxide) on intracellular pH (pHin) and ATP level were evaluated by 31P nuclear magnetic resonance on isolated rat liver submitted to 1 hour of warm ischemia and reperfusion. At the end of the reperfusion period, during which pHin recovered to its initial value (7.16 +/- 0.03) in all groups, the ATP recovery level (expressed in percentage of initial value) was similar in controls and DEPMPO (60% +/- 5%, n = 6 and 54% +/- 4%, n = 6, respectively), but only 37% +/- 1% in DMPO-treated livers (n = 6) (p < 0.05 versus controls and p < 0.05 versus DEPMPO). Oxidative phosphorylation was not affected by an addition of nitrones on isolated mitochondria extracted from livers not submitted to ischemia-reperfusion. In contrast, mitochondria extracted at the end of the ischemia-reperfusion showed an impairment in the phosphorylation parameters, particularly in the presence of DMPO. Mass spectrum of ischemic liver perchloric acid extracts evidenced probable catabolites in treated groups. The differences in the effect of the two nitrones on energetic metabolism may be explained by the production of deleterious catabolites by DMPO as compared to DEPMPO. Even though a specific radical scavenging effect could be operative in the liver, our results indicate that catabolic effects were predominant. The absence of deleterious effects of DEPMPO in contrast to DMPO on the liver energetic metabolism was evidenced, allowing the use of DEPMPO for ESR detection. PMID- 9219182 TI - Removal of the outer lines of the citrate multiplet in proton magnetic resonance spectra of the prostatic gland by accurate timing of a point-resolved spectroscopy pulse sequence. AB - Proton MR spectra of a healthy human prostatic gland show a major signal for citrate appearing as an AB-type multiplet. After application of multipulse localization sequences, the outer lines of this multiplet often appear with dispersion line shapes disturbing the baseline and interfering with proper quantification of citrate itself and other nearby resonances. Based upon analytical descriptions of the time evaluation of an AB spin system during a point resolved spectroscopy (PRESS) pulse sequence (90x-tau 1-180y-tau 2-180y-t), equations were derived representing the intensity of the absorption and dispersion line shape of the outer lines of the citrate multiplet at the top of echo t = tau 2-tau 1. From these equations, it was calculated that the outer lines of citrate can be removed almost completely using a PRESS pulse sequence with tau 1 = 11 ms and tau 2 = 60 ms. The theoretical description was confirmed by the almost complete disappearance of the two outer citrate resonances in in vitro and in vivo proton MR spectra acquired with this pulse sequence timing. PMID- 9219181 TI - Magnetic resonance imaging of the female pelvic floor and urethra: body coil versus endovaginal coil. AB - The anatomy of the female pelvic floor and urethra is complex. With the introduction of MRI, the discussion about the normal anatomy of this area has not diminished. The use of a body coil may be contributary to this. In the present study images obtained with an endovaginal coil are compared with those of a quadrature body coil series to study the possible advantage of endovaginal imaging. Axial and radial T2-w TSE images at a 1.0-T machine were obtained in seven healthy volunteers. The pelvic floor structures as well as the levator ani muscle and the urogenital diaphragm are excellently demonstrated with the endovaginal coil. Also, the urethrovaginal sphincter could be recognized in six volunteers, but only in three with the body coil. In six volunteers a new ligamentous structure, the urethropelvic sling, connecting the urethra to the levator ani muscle and contributing to the supporting mechanism of the urethra is shown with the endovaginal coil. The zonal anatomy of the urethra is excellently shown with the endovaginal coil. The urethral length could only be accurately measured with this coil and ranged from 3.1 to 3.6 cm. Compared with the body coil, endovaginal MRI is excellent in demonstrating the anatomy of the pelvic floor and urethra. PMID- 9219183 TI - Can magnetization transfer magnetic resonance imaging follow proteoglycan depletion in articular cartilage? AB - In this study we determined the efficiency of magnetization transfer magnetic resonance imaging (MT-MRI) to differentiate native and enzymatically degraded cartilage, using bovine sesamoid bones from the metacarpophalangeal joint as a model system. Gradual proteoglycan (PG) depletion was achieved by increasing incubation periods with testicular hyaluronidase. For native cartilage a Ms/Mo ratio of 0.303 +/- 0.09 (mean +/- SEM) was measured. Biochemically determined PG diminution up to 50% correlated strongly (r = 0.953) with changes in the Ms/Mo ratio. Further PG loss is not reflected in an equally drastic Ms/Mo increase, whereas subsequent treatment of PG-depleted cartilage samples with collagenase led to an additional rise in the Ms/Mo ratio. Proteoglycan depletion and the beginning destruction of the collagen structure were also assessed histochemically. Our study confirms that collagen contributes to the baseline MT effect observed in articular cartilage. However, the changes in the MT ratio in gradually PG-depleted cartilage with a largely intact collagen network indicate that PG contributes to the MT effect as well. Therefore MT-MRI might become a sensitive technique for the monitoring of subtle degradational changes in articular cartilage, the still inaccessible process in osteoarthritis. PMID- 9219184 TI - Preliminary experience with a new double-echo half-Fourier single-shot turbo spin echo acquisition in the characterization of liver lesions. AB - A new double-echo half-Fourier single-shot turbo spin echo technique has been implemented in which two images are obtained per excitation pulse, one with an echo time (TE) of 60 ms and another with a TE of 438 ms. The acquisition window per image is 380 ms and is determined by the echo spacing of 4.3 ms and the echo train length of 88 for images with resolution of 160 x 256. No breath holding was performed. The aim of the study was to test whether the additional information of the late TE image improves the characterization of liver lesions. Twenty-eight patients with 39 focal liver lesions (9 cysts, 11 hemangiomas, and 19 solid lesions) were imaged with the new technique, and signal intensity (SI) ratios of lesion and liver were obtained. A t-test analysis showed that in the TE 60 ms image, SI ratios of cysts and hemangiomas were not significantly different, whereas in the TE 438 ms images the two types of lesions can be classified. Signal intensity ratios of solid lesions were in both images clearly lower than those of cysts and hemangiomas. The technique, therefore, seems a promising and straightforward new tool for the characterization of liver lesions. PMID- 9219185 TI - Representation and analysis of medical decision problems with influence diagrams. AB - Influence diagrams are a powerful graphic representation for decision models, complementary to decision trees. Influence diagrams and decision trees are different graphic representations for the same underlying mathematical model and operations. This article describes the elements of an influence diagram, and shows several familiar decision problems represented as decision trees and as influence diagrams. The authors also contrast the information highlighted in each graphic representation, demonstrate how to calculate the expected utilities of decision alternatives modeled with an influence diagram, provide an overview of the conceptual basis of the solution algorithms that have been developed for influence diagrams, discuss the strengths and limitations of influence diagrams relative to decision trees, and describe the mathematical operations that are used to evaluate both decision trees and influence diagrams. They use clinical examples to illustrate the mathematical operations of the influence-diagram evaluation algorithm; these operations are arc reversal, chance node removal by averaging, and decision node removal by policy determination. Influence diagrams may be helpful when problems have a high degree of conditional independence, when large models are needed, when communication of the probabilistic relationships is important, or when the analysis requires extensive Bayesian updating. The choice of graphic representation should be governed by convenience, and will depend on the problem being analyzed, on the experience of the analyst, and on the background of the consumers of the analysis. PMID- 9219186 TI - Use of influence diagrams to structure medical decisions. AB - Influence diagrams are compact representations of decision problems that are mathematically equivalent to decision trees. The authors present five important principles for structuring a decision as an influence diagram: 1) start at the value node and work back to the decision nodes; 2) draw the arcs in the direction that makes the probabilities easiest to assess; 3) use informational arcs to specify which events will have been observed at the time each decision is made; 4) ensure that missing arcs reflect intentional assertions about conditional independence and the timing of observations; and 5) ensure that there are no cycles in the influence diagram. They then build an influence diagram for the problem of staging non-small-cell lung cancer as an illustration. Influence diagrams offer several strengths for structuring medical decisions. They represent graphically and compactly the probabilistic relationships between parameters in the model. Influence diagrams also allow the model to be structured in a fashion that eases the necessary probability assessments, regardless of whether the assessments are based on available evidence or on expert judgment. Influence diagrams provide an important complement to decision trees, especially for representing probabilistic relationships among variables in a decision model. PMID- 9219187 TI - Dollars may not buy as many QALYs as we think: a problem with defining quality-of life adjustments. AB - The scale of health state quality that should be used to compute quality-adjusted life years (QALYs) ranges from 0 (death) to 1.0 (excellent health); this is called the "Q" scale. But many cost-utility analyses (CUAs) in the literature use the upper anchor of the scale to denote only the absence of the particular health condition under investigation, and weight the disease state proportional to this endpoint; these are called "q" scales. Computations using q-scale health-state weights ignore the fact that the average patient is still subject to chronic and acute conditions comorbid with the condition being analyzed; the absence of a particular condition is not in general the same as excellent health, i.e., the Q scale is longer than a q scale. CUAs based on q scales yield "qALYs." Incremental $/qALY ratios are generally lower than $/QALY ratios; in the example presented, $/qALY must be inflated by about 15% to yield $/QALY. Other CUAs correctly weight disease states using the Q scale, but erroneously assign a quality weight of 1.0 to absence of the disease in the CUA computations. The results of such analyses are called "NP-QALYs," as the correction factor to compute QALYs is not a simple proportional adjustment. The authors suggest that analysis doing cost-utility analyses without access to primary data from treated patients use average age specific health-related quality-of-life weights from population-based studies to represent the state of not having a particular disease. Consumers of CUAs should closely examine the nature of the QALYs in any published analyses before making decisions based on their results. PMID- 9219188 TI - The use of confidence intervals for individual utilities: limits to formal decision analysis for treatment choice. AB - This paper discusses the use of confidence intervals for utility measurements. Classic test theory is applied to estimate confidence intervals for utilities. The theory is enhanced to calculate confidence areas for combined utilities and confidence bands for the threshold line. As an example it is shown that, if confidence intervals are taken into account, the implied preferred treatment of T3-larynx carcinoma patients is uncertain for a wide range of utilities, considering the mediocre reliability of most methods of utility assessment. This implies that although utility measurement and formal decision analysis can be a useful way to look at the decision problem, ambiguity, which must be resolved by other means, will often remain. PMID- 9219189 TI - Cardiologists' use of clinical information for management decisions for patients with unstable angina: a policy analysis. AB - Previous studies of management of unstable angina have revealed substantial differences in management between different hospitals, especially with respect to the use of coronary angiography. Physicians in a hospital with angiography facilities were more inclined to perform angiography than were physicians in hospitals without these facilities, even when differences in patient populations were taken into account. The authors compared the management strategies of 18 cardiologists, working in hospitals with and without angiography facilities, using a series of paper-case summaries, in order to assess the contribution of individual variability between physicians to practice differences. Physicians who worked in a hospital with in-house angiography facilities were more inclined to request angiography in similar case summaries, but the inter-individual variation exceeded the between-hospital variation. The variation in individual policies with respect to the decision to initiate coronary angiography could be associated with differences in weighting clinical information. These results confirm that practice variations may have many causes: variability in patients' characteristics, variations in how physicians react to these, differences in the availability of services, and variability in thresholds for action. PMID- 9219190 TI - Patient participation in deciding breast cancer treatment and subsequent quality of life. AB - This investigation of patients with early breast cancer examined relationships among patient involvement in deciding treatment (i.e., whether to undergo breast removal or breast conservation), perceptions of control over treatment decisions, and subsequent health-related quality of life. It was predicted 1) that patients who more actively participated in consultations to decide treatment would perceive more decision control than would more passive patients and 2) that patients who perceived greater decision control would report better health related quality of life following treatment than would patients perceiving less decision control. Sixty patients with stage I or II breast cancer allowed their consultations with surgeons to be audiorecorded. Following these visits, patients reported on their involvement in the consultation, optimism for the future, knowledge about treatment, and two aspects of perceived decision control, the perception of having a choice for treatment and the extent to which the doctor or patient was responsible for the decision. Six and 12 months postoperatively, 51 patients (85%) returned a follow-up survey assessing perceived decision control and health-related quality of life. The first prediction received some support. The patients who had more actively participated in their consultations, particularly in terms of offering opinions, assumed more responsibility for treatment decisions during the year following surgery than did less expressive patients. Also, the patients who reported more involvement in their consultations later believed they had had more of a choice for treatment. The second hypothesis was partially supported. Six and 12 months following treatment, the patients who believed they were more responsible for treatment decisions and believed they had more choice of treatment reported higher levels of quality of life than did the patients who perceived themselves to have less decision control. However, perceived control at the time of treatment did not predict later quality of life. Theoretical and clinical implications are discussed. PMID- 9219191 TI - Making treatment decisions with HIV infection: a pilot study of patient preferences. AB - The importance of understanding patient preferences in making treatment decisions is widely recognized. This pilot study utilized a forced-choice paired-comparison method in which 28 ambulatory HIV-infected patients were given a computer generated presentation of all possible pairs of eight different treatment options for their disease (FDA-approved medications, experimental and alternative treatments, no medication). Preferences were analyzed using binary multidimensional scaling analyses to determine the utility of paired-comparison models for the study of treatment-decision making and to identify factors influencing patient decision making. Results indicated that a three-dimensional model provided the best fit for the data. One dimension correlated with medications that raise CD4+ lymphocyte counts (r = 0.92, p < 0.001) and a second dimension correlated with frequency of dosing (r = 0.97, p < 0.0001). Patients' internal consistency of decision making was inversely correlated with severity of AIDS dementia symptoms as measured by performance on a neuropsychological test battery (r = -0.55, p < 0.0025). This finding indicates that AIDS dementia may significantly hinder patients' ability to use a rational (internally consistent) decision-making strategy in making treatment choices. Results also suggested that AIDS patients base treatment decisions primarily on the likelihood of raising CD4+ cell counts and restrictiveness of dosing regimens, but are not influenced by FDA approval status, volume of empirical support for the medications, or even possible harmful side effects. The implications of these findings for the treatment of patients with AIDS are discussed. PMID- 9219192 TI - Community-based research--a framework for problem formulation: the case of upper endoscopy for gastroesophageal reflux disease. AB - OBJECTIVE: To identify clinical hypotheses and information gaps underlying disagreement about the use of upper gastrointestinal endoscopy (EGD) for the diagnosis of gastroesophageal reflux disease (GERD), and to design a registry study to test these hypotheses. DESIGN AND SETTING: Structured group discussions with community-based practicing gastroenterologists. RESULTS: Thirty-three gastroenterologists from 17 sites discussed a set of clinical scenarios concerning the use of EGD in GERD patients with different clinical histories. Clinicians identified patient characteristics and outcome variables missing from the original problem formulation. Using decision tables, the combinations of patient characteristics that provoked disagreement among clinicians were determined. The resulting decision tables specified which characteristics and outcome variables should be measured to test competing clinical theories of when to use EGD in patients with GERD. Subsequently, the clinicians conducted a practice-based study measuring uncertain variables associated with disagreement about the need for EGD in specific clinical situations. CONCLUSION: A structured, but flexible, approach to group discussion may help identify factors that are important in decision making and the hypotheses that should be addressed in resolving variations in practice styles. Technology assessors can use these methods to identify variables underlying clinicians' concerns about the clinical validity of recommendations about practice. This experience with eliciting patient characteristics and uncertain variables underscores the importance of involving practicing clinicians in the process and could be a useful model for problem formulation in guideline development and in community-based research. PMID- 9219193 TI - Preference values for visual states in patients planning to undergo cataract surgery. AB - To assess how preference values that cataract surgery patients assign to their preoperative visual states relate to visual acuity and problems in specific aspects of daily life, the authors interviewed 47 patients scheduled to have cataract surgery. Using a rating-scale technique with a scale from 0 (death) to 1 (excellent health), the patients had a mean preference value of 0.68 for their preoperative vision. Patients' preference values for their preoperative vision were more closely related to problems in specific aspects of daily life (especially feelings of depression and problems interacting with people) than to visual acuity in the operative eye, better eye, or worse eye, or a weighted average of visual acuities in both eyes. These results provide a rationale for relying more on patients' views about the effects of visual impairment than on measures of visual acuity when assessing the need for cataract surgery. PMID- 9219194 TI - Exorcising protocol-induced spirits: making the clinical trial relevant for economics. AB - Economic evaluation frequently depends on estimates from clinical trials of both effectiveness of treatment and resource utilization accompanying it. Protocol driven events in the trial among other influences often imply that both estimates will be inaccurate. This paper indicates how one may supplement a trial with additional data to connect the artificial trial to the real world of clinical practice. It also shows that data required for this model may be estimated from other sources (via Bayesian modeling) if they are not directly available. The required data include (for example) the proportion of patients with disease who would have presented with clinical signs if they had not been part of a trial that allowed early detection and treatment based on subclinical testing mandated by trial protocol. Those presenting with clinical signs would use additional resources for treatment and/or confirmatory diagnostics. Those with subclinical disease either 1) would never use resources in the case where they never developed clinical manifestations or 2) would use resources of a different type or intensity and perhaps have different outcomes by virtue of their disease's being discovered at a later point in time. Basing resource use and ultimate effectiveness on this revised measure of outcome rather than the one in the trial should lead to more accurate predictions for economic purposes. PMID- 9219196 TI - Influence diagrams: a new dimension for decision models. PMID- 9219195 TI - Incorporating risk attitude into Markov-process decision models: importance for individual decision making. AB - Most decision models published in the medical literature take a risk-neutral perspective. Under risk neutrality, the utility of a gamble is equivalent to its expected value and the marginal utility of living a given unit of time is the same regardless of when it occurs. Most patients, however, are not risk-neutral. Not only does risk aversion affect decision analyses when tradeoffs between short and long-term survival are involved, it also affects the interpretation of time tradeoff measures of health-state utility. The proportional time tradeoff under- or overestimates the disutility of an inferior health state, depending on whether the patient is risk-seeking or risk-averse (it is unbiased if the patient is risk neutral). The authors review how risk attitude with respect to gambles for survival duration can be incorporated into decision models using the framework of risk-adjusted quality-adjusted life years (RA-QALYs). They present a simple extension of this framework that allows RA-QALYs to be calculated for Markov process decision models. Using a previously published Markov-process model of surgical vs expectant treatment for benign prostatic hypertrophy (BPH), they show how attitude towards risk affects the expected number of QALYs calculated by the model. In this model, under risk neutrality, surgery was the preferred option. Under mild risk aversion, expectant treatment was the preferred option. Risk attitude is an important aspect of preferences that should be incorporated into decision models where one treatment option has upfront risks of morbidity or mortality. PMID- 9219197 TI - Health state utility anchors: being clear on what "1" means. PMID- 9219198 TI - Patient participation and decision control: are patient autonomy and well-being associated? PMID- 9219200 TI - Impact of age and gender on peak oxygen uptake in chronic heart failure. AB - Anthropometric and demographic characteristics are important determinants of exercise performance in healthy subjects, but their influence has not yet been studied in severe chronic heart failure, although peak oxygen uptake is frequently assessed in such patients for prognostic purposes. The aim of the present analysis was to examine the association between peak oxygen uptake and age, gender, and measures of body size in patients with severe chronic heart failure. We selected 122 (99 male) adult heart transplant candidates who were able to perform a bicycle ergometer test with respiratory gas analysis until voluntary fatigue. Peak oxygen uptake was higher in male than in female patients, both before and after adjustment for weight. In single regression analysis on the total study population, peak oxygen uptake was positively related to weight and to height, but inversely to age (r = 0.59 (P < 0.001), 0.42 (P < 0.001), and 0.33 (P < 0.001), respectively). Multiple stepwise regression analysis identified weight (P < 0.001), age (P < 0.001), and gender (P < 0.01) as independent determinants of peak oxygen uptake (cumulative R2 = 0.45). Similar to the findings in healthy subjects, peak oxygen uptake of patients with severe chronic heart failure is influenced by anthropometric and demographic characteristics. PMID- 9219199 TI - Effect of acute bicarbonate administration on exercise responses of COPD patients. AB - Patients with severe chronic obstructive pulmonary disease (COPD) are limited in their exercise tolerance by the level of ventilation (VE) they can sustain. We determined whether acutely increasing blood bicarbonate levels decreased acid stimulation to the respiratory chemoreceptors during exercise, thereby improving exercise tolerance. Responses were compared with those obtained during 100% O2 breathing (known to reduce VE in these patients) and to the responses of healthy young subjects. Participants were six patients with severe COPD (forced expired volume in 1 s = 31 +/- 11% predicted) but without chronic CO2 retention and 5 healthy young subjects. Each subject performed three incremental cycle ergometer exercise tests: 1) control, 2) after ingestion of 0.3 g.kg-1 of sodium bicarbonate and 3) while breathing 100% O2. During these tests VE was measured continuously and arterialized venous blood (patients) or arterial blood (healthy subjects) was sampled serially to assess acid base variables. Bicarbonate loading increased standard bicarbonate by 4-6 mmol.L-1 and this elevation persisted during exercise. In both groups, bicarbonate loading resulted in a substantially higher arterial pH; arterial PCO2 was either unchanged (healthy subjects) or mildly (averaging 5 torr) higher (COPD patients). However, in neither group did bicarbonate loading result in an altered VE response to exercise or an increase in exercise tolerance. In contrast, superimposing hyperoxia on bicarbonate ingestion yielded, on average, 24% reduction in VE and 50% increase in peak work rate in the patients (but not in the healthy young subjects). We conclude that acute bicarbonate loading is not an ergogenic aid in patients with severe COPD. PMID- 9219202 TI - Thermoregulatory responses to cycling with and without a helmet. AB - This study examined the effects of wearing a helmet on selected body temperatures and perceived heat sensation of the head and body while cycling in a hot-dry (D) (35 degrees C, 20% relative humidity (RH) and hot-humid (H) (35 degrees C, 70% RH) environment. Ten male and four female cyclists (mean +/- SD: males = age 27 +/- 7 yr, peak O2 uptake (VO2) 4.10 +/- 0.54 L.min-1; females = age 26 +/- 3 yr, peak O2 uptake (VO2) 3.08 +/- 0.49 L.min-1) performed four randomized 90-min cycling trials at 60% of peak VO2 both with (HE) and without (NH) a commercially available cycling helmet in both D and H environments. VO2, core (Te), skin (Tsk), and head skin temperatures, heart rate (HR), rating of perceived exertion (RPE), and perceived thermal sensation of head (TSH) and body (TSB) were measured throughout exercise. For all measured variables, no significant difference was evident between HE and NH. However, Tc, Tsk, and mean head skin temperatures were higher (P < 0.001) in H than D. Likewise, RPE, TSH, TSB (P < 0.001), and sweat rates (H = 1.33 +/- 0.32, D = 1.14 +/- 0.23 L.h-1) (P < 0.01) were higher in H versus D. Results indicate that use of a commercially available cycling helmet while riding in a hot-dry or hot-humid environment does not cause the subjects to become more hyperthermic or increase perceived heat sensation of the head or body. PMID- 9219201 TI - Diaphragm structure and function in health and disease. AB - The diaphragm is the primary muscle of inspiration, and as such uncompromised function is essential to support the ventilatory and gas exchange demands associated with physical activity. The normal healthy diaphragm may fatigue during intense exercise, and diaphragm function is compromised with aging and obesity. However, more insidiously, respiratory diseases such as emphysema mechanically disadvantage the diaphragm, sometimes leading to muscle failure and death. Based on metabolic considerations, recent evidence suggests that specific regions of the diaphragm may be or may become more susceptible to failure than others. This paper reviews the regional differences in mechanical and metabolic activity within the diaphragm and how such heterogeneities might influence diaphragm function in health and disease. Our objective is to address five principal areas: 1) Regional diaphragm structure and mechanics (GAF). 2) Regional differences in blood flow within the diaphragm (WLS). 3) Structural and functional interrelationships within the diaphragm microcirculation (DCP). 4) Nitric oxide and its vasoactive and contractile influences within the diaphragm (MBR). 5) Metabolic and contractile protein plasticity in the diaphragm (SKP). These topics have been incorporated into three discrete sections: Functional Anatomy and Morphology, Physiology, and Plasticity in Health and Disease. Where pertinent, limitations in our understanding of diaphragm function are addressed along with potential avenues for future research. PMID- 9219203 TI - Heart rate performance curve during incremental cycle ergometer exercise in healthy young male subjects. AB - In 1992 Conconi et al. (20) presented an indirect and noninvasive method for the determination of anaerobic threshold (AnT) in an incremental field test for runners. This noninvasive method for the determination of anaerobic threshold is dependent on the occurrence of a deflection of the heart rate performance curve (HRPC). The aim of our study was to evaluate the degree and direction of the deflection of the HRPC and the relationship of the heart rate threshold (HRT) to the lactate turn point in a group of 227 healthy young subjects (age: 23 +/- 4 yr). The subjects were divided into three groups by means of second degree polynomial fitting (GI: regular deflection, kHR > 0.1; G II: no deflection, 0 < kHR < 0.1; G II: inverse deflection, k < -0.1). No significant differences between the groups were found in the anthropometric data or in the power output and the blood lactate concentration at both the first (LTP1) and second (LTP2) lactate turn points and at maximum performance (Pmax). Using the method of Conconi et al. (20), 85.9% of the subjects showed a "regular" deflection, 6.2% showed no deflection at all, and 7.9% showed even an inverted deflection of the HRPC. An HRT could be obtained in both G I and G III, and power output at HRT was not significantly different in comparison to that at the LTP2. No HRT could be assessed in G II. The heart rate at HRT and the LTP2 were significantly lower in G III compared with G I. The phenomenon of heart rate break point may be attractive in training regulation, but its application is limited because a heart rate deflection cannot be found even in young subjects in some cases. PMID- 9219204 TI - Beta 1-adrenoreceptors regulate resting metabolic rate. AB - This was a randomized, cross-over experiment designed to determine which beta adrenergic receptors, beta 1, beta 2, or both, regulate metabolic rate in humans. All subjects (3 women, 4 men) were administered a 7-d therapeutic dose of a selective beta 1-antagonist (atenolol 50 mg BID), a combined beta 1, beta 2 antagonist (propranolol 80 mg BID), and a placebo control (BID). Indirect calorimetry was determined before and after 1 h of submaximal exercise. Exercise was performed at 50% of the trial specific VO2peak because maximal exercise was significantly decreased in the presence of the nonselective beta 1, beta 2 antagonist (VO2peak placebo: 44.90 +/- 4.40 mL.kg-1.min-1 vs beta 1, beta 2 antagonism: 39.20 +/- 3.00 mL.kg-1.min-1; P < 0.05). Both the beta 1 and the combined beta 1, beta 2-adrenoreceptor antagonists reduced resting oxygen consumption to a similar extent (0.247 +/- 0.007 L.min-1 placebo, vs 0.218 +/- 0.007 L.min-1 beta 1-antagonism, vs 0.226 +/- 0.007 L.min-1 beta 1, beta 2 antagonism; P < 0.05). However, the 30-min and 60-min excess post-exercise oxygen consumption (mean EPOC) remained unchanged. It is concluded that the beta 1 receptors are regulating the effects of the sympathetic nervous system on resting but not exercise recovery metabolic rate. These metabolic side effects may suggest that changes need to be made in the nutritional requirements of patients using beta-adrenergic antagonists. PMID- 9219205 TI - Oxygen consumption of cycle ergometry is nonlinearly related to work rate and pedal rate. AB - The purpose of the study was to develop an equation to predict the oxygen cost of cycle ergometry. Forty subjects performed an incremental cycle ergometer test on three occasions at 50, 70, or 90 rpm in a counterbalanced order. Work rate was incremented every 5 or 6 min when steady rate values were achieved. To ensure accurate work rates, ergometer resistance was calibrated and flywheel revolutions were electronically measured. Oxygen consumption was measured with a computer interfaced system which provided results every minute. Oxygen consumption (mL.min 1) was the dependent variable, and independent variables were work rate (WR in kgm.min-1), pedal rate (rpm), weight (Kg), and gender (males, 0; females, 1). The following nonlinear equation was selected; VO2 = 0.42.WR1.2 + 0.00061.rpm3 + 6.35.Wt + 0.1136.RPM50.WR-0.10144.RPM90-WR-52-Gender, R2 = 0.9961, Sy.x = 106 mL.min-1, where RPM50: 50 rpm = 1, and RPM90: 90 rpm = 1, else = 0. It was concluded that the oxygen cost of cycle ergometry is nonlinearly related to work rate and pedal rate, linearly related to weight, and that females use less oxygen for a particular work rate. PMID- 9219206 TI - Predictive accuracy of bioimpedance in estimating fat-free mass of African American women. AB - The purpose of this study was to identify the BIA (bioimpedance analysis) equation that yields the best estimate of body composition for 122 premenopausal African-American women (18-40 yr). Total body density (Db) was determined by hydrodensitometry at residual lung volume and converted to %BFHD using the Siri (31) formula, %BFHD was used to calculate reference fat-free mass (FFM). Resistance and reactance were measured using a Valhalla bioimpedance analyzer. The predictive accuracy of generalized, age-gender, race-specific, fatness specific, and the Valhalla manufacturer's BIA equations was compared. There were significant correlations between FFMHD and FFMBIA for all BIA equations (r = 0.85 to 0.92). Except for the modified Segal fatness-specific equations, the prediction errors (SEE and E) exceeded 2.8 kg. For individuals, the %BF derived from FFMBIA predicted by the modified Segal equations was within +/- 3.5% BF for 69% of the subjects. This percentage was less (34-53%) for other equations. These results suggest that the predictive accuracy of BIA for estimating body composition of African-American women is improved when fatness-specific equations are used. We recommend using the modified Segal fatness-specific equations to assess FFM and %BF of premenopausal African-American women. PMID- 9219207 TI - Physical activity, sports participation, and risk factors in adolescents. AB - The purpose of this study was to analyze the relationships between physical activity (ACT), including sports participation (SP) and antecedent risk factors for coronary heart disease (CHD), in a representative sample of adolescents from Northern Ireland, a region of high coronary mortality. Biological and behavioral risk factors were measured in a random sample of 1015 school children aged 12 and 15 yr. ACT and SP were assessed by self-report questionnaire, and relationships with biological risk factors were analyzed with stepwise multiple linear regression after controlling for potential confounders. Results showed that in 15 yr-old males ACT was beneficially associated with systolic blood pressure (P < 0.05), lipid profile, and cardiorespiratory fitness (both P < 0.01). In 15-yr-old females, SP was associated beneficially with fatness and cardiorespiratory fitness. Odds ratios calculated from logistic regression revealed that for the older children, a relatively small drop (-20%) in ACT (boys) or SP (girls) was significantly related to the probability of exposure to multiple risk factors. Overall, relationships were stronger for males rather than females and for older rather than younger children. This study provides further evidence for beneficial associations between ACT, SP, and CHD risk status in adolescents. PMID- 9219208 TI - Mechanical characteristics of knee extension exercises performed on an isokinetic dynamometer. AB - The purpose of this study was to evaluate selected mechanical characteristics of knee extension exercises performed on a LIDO Active Isokinetic System. A female subject performed two repetitions of maximal effort knee extension at 16 different preset angular velocities (PAVs). The gravitational and inertial effects were included in the computation of the resultant knee torque. For each repetition, the knee flexion angle, the angular velocity and acceleration of the shank, and the knee torque throughout the range of motion were computed. The shank angular acceleration values indicated that if the inertial effect is not considered the knee torque will be underestimated in the initial phase and errors in knee torque up to about 6 N.m can be expected for the rest of the repetition. The durations when the shank angular velocity was within +/- 5% and +/- 10% of PAV (expressed as percentages of the repetition time) were found to decrease with increasing PAV. The difference between PAV and shank angular velocity at the instant of peak torque also increased with increasing PAV. The results demonstrate the limitations that may exist in an isokinetic dynamometers. PMID- 9219209 TI - Low back loads over a variety of abdominal exercises: searching for the safest abdominal challenge. AB - Abdominal exercises are prescribed for both the prevention and treatment of low back injury. However, these exercises sometimes appear to have hazardous effects on the lumbar spine. The purpose of this study was to identify quantitatively abdominal exercises that optimize the challenge to the abdominal muscles (rectus abdominis, external oblique, internal oblique) but impose minimal load penalty to the lumbar spine. Nine volunteers performed 12 different abdominal exercises. For a given task the maximum abdominal muscle EMG value was divided by the maximum compression value, resulting in an abdominal challenge versus spinal compression cost index. In general, the partial curl-ups generated the highest muscle challenge-to-spine cost indices. However, those exercises that generated the best challenge-to-cost indices did not necessarily record the lowest compression levels along with the highest EMG activations. No single exercise was found that optimally trained all of the abdominal muscles while at the same time incurring minimal intervertebral joint loads. It was concluded that a variety of selected abdominal exercises are required to sufficiently challenge all of the abdominal muscles and that these exercises will-differ to best meet the different training objectives of individuals. PMID- 9219210 TI - Physical fitness as a determinant of vagal modulation. AB - The association between increasing age and decreasing vagal modulation is well known. However, the importance of fitness as a determinant of the decline in vagal modulation with age is not established. To test the hypothesis that decreasing vagal modulation is largely a function of declining fitness rather than increasing age, we studied a sample of healthy volunteers with a wide range of fitness levels, but a narrow age range. We assessed fitness by measuring the maximal oxygen uptake (VO2max) achieved during incremental bicycle exercise. Vagal modulation was assessed by calculating high frequency power (0.15-0.40 Hz) of the RR variability power spectrum from 24-h ECG recordings. We studied 37 healthy volunteers who were 22-44 yr old. In our sample, VO2max ranged from 25 to 70 mL.min-1.kg-1 (mean of 45 +/- 13). Age was not significantly related to high frequency power, but VO2max was highly correlated with high frequency power (r = 0.74, P = 0.0001), indicating that physical fitness is strongly associated with vagal modulation. Thus, the decline in vagal modulation often attributed to increasing age may, instead, be the result of a decline in fitness. PMID- 9219212 TI - Cardiovascular responses to submaximal exercise in 7- to 9-yr-old boys and girls. AB - The purpose of this study was to investigate whether differences exist between boys and girls in submaximal cardiovascular responses to exercise on both the treadmill and cycle ergometer. Twenty-four (12 boys and 12 girls) 7- to 9-yr-old children participated in two maximal (one treadmill and one cycle) and four submaximal tests (two treadmill and two cycle). There were no significant differences between the boys and girls in maximal oxygen consumption (L.min-1 or mL.kg-1.min-1) or physical characteristics except for a significantly larger left ventricular mass in the boys versus the girls (78.8 vs 66.0 g, respectively). Submaximal cardiovascular variables were measured at three different work rates on both exercise modalities. Oxygen consumption at the different work rates was not different between boys and girls on either exercise modality. The trend was for heart rate to be lower and stroke volume higher in boys versus girls, but this difference was only significant for heart rate at 4 miles.h-1 (142.9 vs 155.5 beats.min-1, respectively). It is concluded that in this sample of 7- to 9 yr-old boys and girls there are few significant differences in submaximal cardiovascular responses to exercise on either exercise modality. PMID- 9219211 TI - Influence of different racing positions on metabolic cost in elite cyclists. AB - The spectacular improvements of the 1-h world record in cycling in the last four years have highlighted the importance of aerodynamics in modern bicycle racing. We have investigated the metabolic consequences of the low-crouched aero positions necessary to reduce air drag. In this study, 14 elite male bicycle racers (24.0 +/- 1.0 yr, VO2max 69.4 +/- 0.5 mL.kg-1.min-1) were tested for oxygen consumption (VO2) and heart rate (HR) at 70% (302.6 +/- 5.3 W) of their individual VO2max in three different riding positions during a single test run. The subjects rode their racing bicycles on a wind braked roller; the sequence of the three following positions was randomized: 1) upright cycling (UP), cadence 90 rpm; 2) hands on drops (DP), 90 rpm; and 3) hands on clip-on aero-handlebars (AP), 90 rpm. VO2 and HR values in AP were significantly higher by 1.5 mL.kg 1.min-1 and 5 beats.min-1, respectively, compared with UP. We concluded that riding a bicycle in an extreme aero-position increases the metabolic cost of cycling when wind resistance is not taken into account. However, when the mechanical power losses of 9 W (estimated by the VO2 increase) are compared with the expected aerodynamic power savings of approximately 100 W, it appears that aerodynamic advantages by far outweight their metabolic cost. PMID- 9219213 TI - Ramp and constant power trials produce equivalent critical power estimates. AB - The standard critical power test protocol on the cycle ergometer prescribes a series of trials to exhaustion, each at a different but constant power setting. Recently the protocol has been modified and applied to a series of trials to exhaustion each at a different ramp incremental rate. This study was undertaken to compare critical power and anaerobic work capacity estimates in the same group of subjects when derived from the two protocols. Ten male subjects of mixed athletic ability cycled to exhaustion on eight occasions in randomized order over a 3-wk period. Four trials were performed at differing constant power settings and four trials on differing ramp incremental rates. Both critical power and anaerobic work capacity were estimated for each subject by curve fitting of the ramp model and of three versions of the constant power model. After adjusting for inter-subject variability, no significant differences were detected between critical power estimates or between anaerobic work capacity estimates from any model formulation or from the two protocols. It is concluded that both the ramp and constant power protocols produce equivalent estimates for critical power and anaerobic work capacity. PMID- 9219214 TI - Effect of training on lactate/ventilatory thresholds: a meta-analysis. AB - The purpose of the investigation was to determine the effect of exercise training intensity on the lactate and ventilatory thresholds in sedentary and in active subjects using meta-analysis procedures. The original analyses included 85 study groups from 34 studies. The dependent variable was oxygen consumption at the specified threshold, and the independent variables were training intensity (control and four intensities ranging from below threshold to near maximum) and fitness level (sedentary and conditioned). Data were analyzed statistically using methods described by Hedges and Olkin (13). The results showed that sedentary subjects (effect size (ES) = 2.32) improved significantly over controls (ES = 0.15), while conditioned subjects (ES = 0.63) showed nonsignificant gains. There were no significant differences among training intensities within the fitness categories (Sed ES = 1.6 - 3.1; Cond ES = 0.3 - 1.1) although the conditioned subjects tended to respond better to high intensity training (ES of 1.1 vs 0.4). It was concluded that training at an intensity near the lactate or ventilatory threshold is an adequate training stimulus for improving the thresholds for sedentary subjects, but a higher intensity may be necessary for conditioned subjects. Detraining will reduce lactate and ventilatory thresholds. PMID- 9219215 TI - A review of the microcirculation of adipose tissue: anatomic, metabolic, and angiogenic perspectives. AB - Adipose tissue microcirculation is unique within the vascular system because of a capacity for this tissue to grow throughout most of adult life. A review of the microcirculation of adipose tissue has included a historical review of the early studies, which served as a foundation for later investigations on this topic, including basic hemodynamic measurements in mammalian adipose tissue. The various methods for measuring blood flow in white and brown adipose tissue are discussed with respect to studies of transport of substrates involved in adipose tissue metabolism. The role of innervation and vascular adrenergic receptors and the effects of diet and exercise on adipose tissue blood flow are also included. An in-depth analysis of the development of adipose tissue microvasculature indicates that angiogenesis often precedes adipogenesis. The clinical effects of hemodynamic adaptations to adipose tissue expansion are discussed in view of an epidemic increase in the prevalence of obesity and its co-morbidities. The recent discovery of sites of nuclear regulation of adipocyte differentiation, together with the identification of growth factors in adipose tissue, is an indication of the progress that is being made in the further understanding of molecular and cellular events that affect adipose tissue growth and, ultimately, adipose tissue microcirculation. PMID- 9219216 TI - Blood flow in the cerebral capillary network: a review emphasizing observations with intravital microscopy. AB - Capillary perfusion in the brain is characterized by an essentially continuous flow of erythrocytes and plasma in almost all capillaries. Rapid fluctuations and spatial heterogeneity or red blood cell (RBC) velocity (0.5-1.8 mm/s) within the capillary network are present. In addition, low-frequency (4-8 cpm) synchronous oscillations in RBC velocity in the capillary network emerge when perfusion to cerebral tissue is challenged. Despite the tortuous, three-dimensional architecture of microvessels, functional intercapillary anastomoses are absent. At rest, red cells travel through the capillary network in 100-300 ms along 150- to 500-micron-long paths. Physiological challenges elicit sizable changes in RBC velocity with a minor role for capillary recruitment, change in capillary diameter, or flow shunting. During acute hypoxia, RBC velocity increases in all capillaries; the corresponding response to hypereapnia is more complex and involves redistribution of capillary flow toward more homogeneous perfusion. The response of capillary flow to decreased perfusion pressure reflects autoregulation of cerebral blood flow but also involves intranetwork redistribution of RBC flow between two populations of capillaries, postulated as thoroughfare channels and exchange capillaries. Flow reserve may be provided by the thoroughfare channels and may help maintain flow velocity and capillary exchange and protect the microcirculation from perfusion failure. Isovolemic hemodilution increases RBC velocity three- to fourfold and increases RBC flux to a moderate degree with a relatively small decrease in capillary hematocrit, under normal and compromised arterial blood supply. In cerebral ischemia, leukocyte adhesion is enhanced and appears reversible when the ischemia is moderate but may be progressive when the injury is severe. The observed flow behavior suggests the presence of a physiological regulatory mechanism of cerebral capillary flow that may involve communication among various microvascular and parenchymal cells and utilize locally acting endothelial and parenchymal mediators such as endothelium derived relaxing factor or nitric oxide. PMID- 9219217 TI - Femoral artery ligation stimulates capillary growth and limits training-induced increases in oxidative capacity in rats. AB - OBJECTIVE: The purpose of this study was to assess the interaction of arterial insufficiency and exercise training on soleus and plantaris muscle capillarity and oxidative capacity in adult rats. METHODS: Arterial insufficiency was created by ligation (LIG) of the right femoral artery, and exercise training (TR) was performed on a rodent treadmill. The left hindlimb served as a normally (NORM) perfused control. Capillary:fiber ratio number of capillary contacts per fiber, and citrate synthase activity (CS) were evaluated in the plantaris (Plant) and soleus (Sol) muscles. RESULTS: In sedentary rats, CS was similar between LIG and NORM (Plant: 24.4 vs. 24.3 mumol.min-1.g-1; Sol: 16.6 vs. 16.9 mumol.min-1.g-1), but capillaries per fiber and capillary contacts per fiber were significantly elevated in the plantaris muscle of LIG (2.46 vs. 2.10 caps/fiber, 5.78 vs. 5.03 capillary contacts). CS was elevated in both limbs of TR but was lower in LIG than in NORM (Plant: 28.5 vs. 32.4 mumol.min-1.g-1; Sol: 21.1 vs. 24.9 mumol.min 1.g-1). Treadmill training did not significantly affect capillarity in NORM. However, muscles in the ligated limb of TR tended to have greater capillarity than comparable muscles in either NORM of TR of LIG in SED. CONCLUSIONS: These results demonstrate capillary proliferation in the plantaris but not soleus muscle of rat hindlimbs with femoral artery ligation. Capillarity and CS adaptations were not obligatorily related in LIG, and femoral artery ligation and exercise training appeared to have interactive effects on skeletal muscle capillarity. PMID- 9219218 TI - Remodeling of capillary network in left ventricular subendocardial tissues induced by intravenous vasopressin administration. AB - OBJECTIVE: The question of whether the coronary vasospasm induced by intravenous administration of vasopressin produces any remodeling of the capillary network in the left ventricle was investigated. To this end, cardiac tissues obtained from vasopressin-injected rats were stained to allow capillary counting and for basic fibroblast growth factor (bFGF). METHODS: Nine male Donryu rats were divided into three groups that received, respectively, 0.25 ml of saline containing 0, 0.5, or 1.0 U/kg vasopressin injected into the tail vein once daily for 4 days. Rats were killed 30 days after the last injection. Two additional rats each received a single intravenous injection of 1.0 U/kg vasopressin and were killed 24 hours later. The left ventricles were removed and 16- or 10-micron frozen sections were cut for differential staining and distribution of bFGF, respectively. Differential staining was used to classify the capillary portions, and bFGF was identified by immunohistological staining. RESULTS: Compared with the control group, total capillary density was increased in both vasopressin-treated groups, capillary to myocyte ratio was increased, and the capillary domain areas decreased in the three capillary portions. Arteriolar and intermediate capillary portions increased, while the venular capillary portion decreased. In rats killed 24 hours after vasopressin injection, a considerable amount of bFGF could be demonstrated immunohistochemically in the ventricular tissues, and the punctate distribution of bFGF was still found in rats killed 30 days after treatment. CONCLUSIONS: A remodeling of capillary network which would increase the oxygen transport capacity to cardiac tissues was produced in left ventricular tissues by intravenous injection of vasopressin. bFGF located around capillaries and in the interstitial space may have been involved in the capillary remodeling. PMID- 9219219 TI - Vascular endothelial cadherin (VE-cadherin): cloning and role in endothelial cell cell adhesion. AB - OBJECTIVE: To identify proteins responsible for intercellular junction integrity in human umbilical vein endothelial cells (HUVEC), we produced a monoclonal antibody that recognized an endothelial cell-specific, junctionally restricted protein. We characterized and cloned the antigen to study its functional properties. METHODS: The size and cellular distribution of the antigen were determined by immunofluorescence and immunoprecipitation. The molecule was cloned and transfected into cell lines, and its role in cell-cell adhesion and growth rate was determined. RESULTS: Monoclonal antibody hec1 recognizes VE-cadherin, an endothelial cell-restricted cell adhesion molecule. VE-cadherin is localized to the borders between apposing endothelial cells but is diffusely distributed on subconfluent or migrating cells. Transfection of fibroblasts with VE-cadherin imparts to them the ability to adhere to each other in a calcium-dependent homophilic manner. Expression of VE-cadherin over a several-log range does not change the growth rate of these cells. CONCLUSIONS: Despite the fact that VE cadherin is a "nonclassical" cadherin by structure, it functions as a classic cadherin by imparting to cells the ability to adhere in a calcium-dependent, homophilic manner. On HUVEC it appears to play a role in maintaining monolayer integrity. PMID- 9219220 TI - E-selectin is upregulated in proliferating endothelial cells in vitro. AB - OBJECTIVE: E-selectin is an endothelial cell-specific membrane glycoprotein that participates in leukocyte adhesion and has also been suggested to function in angiogenesis. To gain further insights into E-selectin, we analyzed E-selectin polypeptide in proliferating versus quiescent bovine capillary endothelial cells and its expression as a function of the cell cycle. METHODS: E-selectin polypeptide was analyzed by immunoadsorption from 35Scysteine-labeled endothelial cells, by enzyme-linked immunosorbent assay, and by fluorescence-activated cell sorting. The distribution of endothelial cells in Gzero/G1, S, and G2/M phases of the cell cycle was determined using propidium iodide staining of DNA. RESULTS: E selectin was upregulated in subconfluent proliferating bovine capillary endothelial cells compared to confluent quiescent cultures. The upregulation was independent of activation in that E-selectin was further increased by treatment with tumor necrosis factor alpha or lipopolysaccharide. In contrast to E selectin, P-selectin and platelet-endothelial cell adhesion molecule-1 did not appear to be regulated by the growth state of the endothelial cells. The distribution of E-selectin-positive cells in GzeroG1, S, and G2/M phases of the cell cycle differed from E-selectin-negative cells in that more of the E-selectin positive cells were in G2 and M. CONCLUSIONS: Increased E-selectin expression under noninflammatory conditions is correlated with cellular proliferation and G2/M phases of the cell cycle. The expression of E-selectin in proliferating endothelial cells in vitro is consistent with the presence of E-selectin in proliferating endothelial cells in vivo (Kraling et al. [18]). PMID- 9219221 TI - Shear rate dependency of red cell sequestration in skin capillaries in sickle cell disease and its variation with vasoocclusive crisis. AB - OBJECTIVE: To develop techniques for assessing the sequestration of red blood cells (RBCs) in skin capillaries of sickle cell disease (SCD) patients due to RBC adhesion to endothelium (EC) and RBC aggregation, and to determine the extent to which these processes correlate with onset of painful vasoocclusive crisis. METHODS: Video recordings of nailfold capillaries in the skin of patients with SCD were made during steady-state periods and episodes of painful crisis. A transient low-flow state was induced with a pressure cuff and reductions in RBC velocity were measured by spatial cross-correlation of light intensity along arterial and venous capillary limbs. An RBC accumulation index (AI) was calculated from RBC flow to represent the percentage of arterial in-flow sequestered in a capillary. An index of hematocrit (HI) was derived from axial distributions of light intensity, and a sequestration index (SI) was calculated to represent the relative increase of venous limb HI relative to that in the arterial limb with onset of stasis. RESULTS: Both AI and SI increased dramatically from zero at steady flow to a maximum as shear rates within the capillary were reduced to zero. The increase was small until shear rates (gamma) fell below a transition value (gamma T), following which both AI and SI increased sharply with onset of stasis. For 20 < or = gamma < gamma T the transient increase in AI was significantly elevated in the order AICRISIS > AISTEADY STATE > AICONTROL, thus reflecting increasing RBC sequestration in the venous limb due to either adhesion or aggregation. Compaction of RBCs in the venous limb was evidenced by increased SI that was greater than control for both steady-state and crisis subjects, but insignificantly elevated during crisis compared to steady state, thus supporting a lesser role of RBC aggregation. CONCLUSIONS: Transient sequestration of sickle RBCs in the low-flow state appears to be dominated by RBC EC adhesion, which becomes enhanced during crisis. Although aggregation may enhance adhesive contact of RBCs with EC, it does not increase to the same extent as the rate of sequestration, thus reflecting a greater role of RBC-EC adhesion. PMID- 9219222 TI - Arteriolar dilation produced by venule endothelium-derived nitric oxide. AB - OBJECTIVE: We conducted bioassay experiments to determine whether nitric oxide produced by endothelial cells (endothelial-derived nitric oxide, or EDNO) within large venules could act to dilate arterioles. METHODS: In these experiments parallel segments of first-order arterioles and venules were isolated from skeletal muscle and were cannulated in series with a glass connecting tube (length: 300-500 microns). Arterioles were mechanically denuded of endothelium by a delicate yet abrasive rubbing technique. Venular endothelium remained intact. Endothelial denudation of arterioles was confirmed by the absence of dilation during exposure to acetylcholine (10(-6) mol/L). The cannulated vessels were pressurized to 30 cm H2O and the arterioles pre-constricted by approximately 50% with norepinephrine (10(-10) mol/L). RESULTS: Topical applications of acetylcholine (10(-6) mol/L) or bradykinin (10(-9) mol/L) during luminal perfusion from venule to arteriole produced significant arteriolar dilation. In contrast, a slight arteriolar constriction was observed when the direction of flow was reversed (i.e., arteriole to venule) in the presence of either acetylcholine (10(-6) mol/L) or bradykinin (10(-9) mol/L). Inhibition of venular EDNO with NG-monomethyl-L-arginine (L-NMMA; 10(-5) mol/L; 1 hour) completely abolished the arteriolar dilation observed in response to acetylcholine or bradykinin during venule to arteriole perfusion. CONCLUSIONS: These results demonstrate that venular-derived EDNO can relax arteriolar vascular smooth muscle. PMID- 9219223 TI - Regulation of E-selectin, P-selectin, and intercellular adhesion molecule 1 expression in mouse cremaster muscle vasculature. AB - OBJECTIVE: To investigate the expression of P- and E-selectin and intercellular adhesion molecule 1 (ICAM-1) in the vasculature of mouse cremaster muscles both with and without tumor necrosis factor alpha (TNF-alpha) treatment. METHODS: Mice received injections of monoclonal antibody to P-selectin, E-selectin, or ICAM-1 before fixation to restrict detection to antigen expressed on the endothelial surface. Whole-mount preparations of mouse cremaster muscles were fixed in acetone and stained using biotinylated secondary antibody and peroxidase conjugated streptavidin. RESULTS: P-selectin is expressed on the endothelial surface of cremaster muscle venules within 10 minutes after exteriorization. Expression increases upon treatment with TNF-alpha (2 hours), reflecting transcriptional regulation of P-selectin expression in situ. The baseline E selectin expression is patchy and barely detectable but shows a significant upregulation after treatment with TNF-alpha. [CAM-1 is constitutively expressed in unstimulated mouse cremaster venules and slightly upregulated after 2 hours of TNF-alpha treatment. Under baseline conditions, neither E-selectin, P-selectin, nor ICAM-1 is detectable in arterioles or capillaries. After TNF-alpha treatment, arterioles stain faintly for P-selectin, but not E-selectin or ICAM-1. CONCLUSION: The temporal and spatial pattern of expression of P- and E-selectin and ICAM-1 is consistent with the functional role of these molecules in mediating preferential leukocyte rolling and adhesion in mouse cremaster muscle venules. PMID- 9219224 TI - Cloning of cold-inducible dehydrin-like genes from the blackcurrant (Ribes nigrum L.) using RT-PCR. AB - A method is described for the identification of conserved genes in one plant species by using sequence information on internal motifs from well-characterized clones from another species. This sequence information is used to design primers for reverse transcriptase polymerase chain reaction (RT-PCR) and to design oligonucleotide probes to identify genuine positive amplification products. The approach was successfully used to clone cDNAs encoding cold-inducible dehydrin like genes from the woody perennial blackcurrant, Ribes nigrum L. The strategy described can accelerate the cloning of heterologous cDNAs and is a convenient alternative to direct screening of cDNA libraries. PMID- 9219226 TI - Digoxigenin labeling. AB - Digoxigenin is increasingly used as a label for nonradioactive detection of nucleic acids and proteins. A variety of methods for labeling are described as well as numerous applications of technology. PMID- 9219225 TI - Quantification of the presence and activity of specific microorganisms in nature. AB - Traditional techniques for assessment of microbial numbers and activity generally lack the specificity required for risk assessment following environmental release of genetically engineered microbial inocula. Immunological and molecular-based techniques, such as DNA probing and genetic tagging, were initially used to determine the presence or absence of microorganisms in environmental samples. Increasingly they are being developed for quantification of populations of specific organisms, either indigenous or introduced, in the environment. In addition, they are being used to quantify the activity of particular organisms or groups of organisms, greatly extending the range of techniques available to the microbial ecologist. This article reviews the use of traditional techniques for the quantification of microbial population size and activity and the application of molecular techniques, including DNA probing, genetic marking, use of fluorescent probes, and quantitative PCR, in combination with advanced cell detection techniques such as confocal laser scanning microscopy and flow cytometry. PMID- 9219228 TI - Gene transfer by biolistic process. AB - Gene transfer into somatic tissues is a tool for both the study of gene function in the basic science laboratory and for gene therapy and genetic immunization in the clinic. Biolistic processes can be used to deliver both viral and nonviral vectors into somatic tissues. This review discusses the advantages and disadvantages of three biolistic processes: jet injection, microparticle bombardment, and needle and syringe injection. Jet injection and needle and syringe injection can be used to deliver both viral and nonviral vectors. Both jet injection and microparticle bombardment can be used to target a broad range of tissues. Needle and syringe injection has been most widely used in muscle tissue. The choice of which biolistic process to use is dependent on the specific application. PMID- 9219227 TI - Ribozymes. Their functions and strategies for their use. AB - The ability to alter genes in order to regulate their expression has become an undeniable reality. This can be performed in vitro and in cells, and the possibility of treating diseases and even preventing them now exists through such gene manipulation. A particularly intriguing form of manipulation that has been investigated for just over a decade is one that involves the use of ribozymes. These are short segments of RNA that form complementary base-pairing with mRNA. However, it is their enzymatic properties that set them apart from other antisense RNA molecules and allow them to cleave and destroy mRNA in a very specific manner. The ribozyme then dissociates from the cleaved substrate RNa, and repeatedly hybridizes to and cleaves additional substrate RNA molecules. Problems being addressed as this technology evolves involve optimization of ribozyme:substrate binding efficiencies and their effective transmission into cells. This article points out the origin of ribozymes, analyzes and summarizes the current strategies for designing ribozymes, and outlines a basic procedure for ribozyme development. PMID- 9219229 TI - The use of monoclonal antibodies with colloidal gold-labeled probes in postembedding immunoelectron microscopy. AB - This article focuses on procedures for the use of monoclonal antibodies (MAb) in immunoelectron microscopy (IEM) for the subcellular localization of antigens or to relate function with structure in prokaryotic and eukaryotic cells using postembedding immunoelectron microscopy techniques. The use of MAbs greatly increases the specificity and quality of information when used in combination with gold-labeled probes. Because of its specificity, the reactivity of MAb may be very sensitive to antigenic changes resulting from the process of sample preparation when performing IEM studies. Specific protocols for each particular combination of epitope/MAb must be usually specifically devised, since it is impossible to predict an experimental system that will successfully preserve structures and antigenic determinants for every combination. In this article, we discuss critical technical aspects that usually result in improved resolution when the procedure is used to identify structure in diverse prokaryotic and eukaryotic cells. PMID- 9219230 TI - The use of RNA probes for the analysis of gene expression. AB - The monomeric bacteriophage RNA polymerases allow the synthesis of virtually any RNA molecule in unlimited quantity. In this protocol, we describe the preparation of plasmid and PCR-derived templates. A basic transcription protocol is provided with several optional modifications. The use of RNA probes in Northern blot hybridization and in RNase protection assays is described. The relative advantages and pitfalls of these two methods to quantitatively detect mRNA targets are discussed. PMID- 9219231 TI - Nucleic acid in situ probing. Hybridization to human chromosomes of an alkaline phosphatase labeled centromeric probe. AB - The main principles that underly the use of nucleic acid probes for in situ hybridization are summarized. These include probe design, target preparation, hybridization formats and conditions, and signal generating systems. These principles underly the specific protocol that is described, namely the use of an akaline phosphatase-labeled cloned sequence of the alphoid repeated DNA family as a centomere probe for human chromosomes. PMID- 9219232 TI - Cryopreservation of mammalian embryos. AB - As an innovative method for embryo cryopreservation, vitrification not only reduced the cooling stage duration to a minimum, but also eliminated any injuries cased by extracellular ice, which is a major cause of cell injury. Therefore, if embryos are treated adequately, high survival can be obtained. As a component of a vitrification solution, a permeating cryoprotective agent is essential, and additional inclusion of a macromolecule and a small saccharide makes the solution more effective. The author's group composed a solution, designated EFS40, with ethylene glycol, Ficoll, and sucrose. This solution proved effective for the cryopreservation of various stages of embryos in many species. In this article, the author describes the detailed procedure for the vitrification of mouse morulae; related information is also described. PMID- 9219233 TI - Site-directed mutagenesis using positive antibiotic selection. AB - Various mutagenesis protocols have been established that use the hybridization of a mismatched oligonucleotide to prime DNA synthesis on an M13 phagemid template. For efficient mutagenesis, all of these methods require a means to select for the mutant strand before or during amplification in an Escherichia coli host. In the Altered Sites II protocol, the mismatched oligonucleotide and an oligonucleotide that restores antibiotic resistance to the phagemid are simultaneously hybridized to the template and coupled by DNA synthesis and ligation. The restored antibiotic resistance is then used to select only those phagemids which incorporate the antibiotic repair oligonucleotide. Generally, between 60 and 90% of the phagemids recovered will incorporate both oligonucleotides. This method provides a simple an efficient technique for introducing specific mutations into DNA. PMID- 9219234 TI - An efficient random mutagenesis technique using an E. coli mutator strain. AB - Random mutagenesis of a cloned gene remains a central method to understand many aspects of the gene products function and structure. Having the ability to introduce a limited number of changes within a gene in a controlled fashion allows one to evaluate single changes and study the effect these variants have on the gene of interest. The in vivo random mutagenesis strategy described in this article, using an E. coli host, is a convenient method to introduce a limited number of mutations in a controlled manner. PMID- 9219235 TI - Decontamination of polymerase chain reaction reagents for detection of low concentrations of 16S rRNA genes. AB - We describe a polymerase chain reaction (PCR) that allowed detection of rRNA consensus sequences from the DNA extracted from a wide range of bacterial species in amounts as low as 10 fg. To avoid false positive results with universal primers for 16S rRNA PCR, contaminating DNA had to be eliminated from the polymerase preparations. Decontamination was undertaken before PCR to optimize treatment with DNaseI, and was followed by DNase inactivation at 94 degrees C for 50 min, which eliminated contaminating DNA at concentrations of up to 100 pg. After optimization of PCR conditions for each polymerase. Deep Vent Exo polymerase (New England Biolabs, Beverly, MA), and super-Taq polymerase (HT Biotechnology, Cambridge, UK) were more effective than Ampli-Taq polymerase (Perkin-Elmer Cetus, Norwalk CT), Ampli-Taq LD polymerase (Perkin-Elmer Cetus) or Deep-vent polymerase (New England Biolabs). The technique described in this article might prove to be a universal method for PCR detection of small numbers of unidentified bacteria in usually sterile clinical sites, such as blood and cerebrospinal fluids, in which a broad spectrum of pathogens can be expected. PMID- 9219236 TI - The quantitative analysis of multiple mRNA species using oligonucleotide probes in an S1 nuclease protection assay. AB - The quantitative measurement of steady-state mRNA levels is fundamental to the analysis of gene expression. A variety of techniques are widely used to achieve this including Northern blotting, RNase protection, and S1 nuclease protection. We describe here in detail a relatively recent extension of the S1 nuclease protection technique (1) in which radiolabeled oligonucleotides are used as probes in a solution hybridization assay (2). The principle advantage of this technique is that it allows, in a single RNA sample, the simultaneous measurement of the relative levels of at least six mRNA species, including that of a control mRNA. Further, a large number of RNA samples can be analyzed at one time. PMID- 9219237 TI - High-level expression of a cDNA for human granulocyte colony-stimulating factor in Chinese hamster ovary cells. Effect of 3'-noncoding sequences. AB - We compared the production of recombinant human granulocyte colony-stimulating factor (rhG-CSF) by Chinese hamster ovary (CHO) cells in a transient expression system, using different analogous vectors carrying a human G-CSF-encoding cDNA under the transcriptional control of the murine cytomegalovirus (CMV) major immediate early promoter. Comparison of two transcription units carrying a human (h)G-CSF cDNA deleted of 3'-untranslated (UTR) sequences containing AT-rich elements (ARE) and using 3'-UTR sequences for processing of transcripts from the SV40 early region or from the rabbit beta 1-globin gene showed that use of the sequences from the rabbit beta 1-globin gene resulted in 7- to 12-fold higher levels of rhG-CSF production. Deletion of ARE of hG-CSF cDNA resulted in increased rhG-CSF synthesis when transcription units using 3'-UTR sequences from the rabbit beta 1-globin gene were compared. By contrast, deletion of ARE did not appear to affect rhG-CSF production when 3'-UTR sequences from the SV40 early region were used. The most efficient G-CSF transcription unit, fused to a dihydrofolate reductase (DHFR) marker gene and transfected into a CHO cell line, yielded initial transfectant CHO cell lines secreting up to 21 micrograms rhG CSF/1 x 10(6) cells in 24 h. After two rounds of DHFR gene amplification, a cell line was isolated that contains approx 12 copies of the vector and produces rhG CSF at a rate of 90 micrograms/1 x 10(6) cells in 24 h. PMID- 9219238 TI - Anti-HIV ribozymes. AB - HIV is an RNA virus that replicates intracellularly through various RNA intermediates. Several of these can be targeted by ribozymes (catalytic RNA molecules), and a number of investigators, including this group, have demonstrated the ability of ribozymes to suppress HIV replication in this way. It is argued that this gene therapy approach may be viewed as an adjunct to chemotherapeutic drugs, which may allow not just viral suppression, but also immune restoration. This can only finally be tested in clinical trials, and several are planned. The basic ribozyme unit, the potential of which was described less than 10 years ago, is about to be tested in an amunable disease state. PMID- 9219240 TI - Impact of molecular biology on the detection of foodborne pathogens. AB - Molecular biological methods that use antibodies and nucleic acids to detect specific foodborne bacterial pathogens were scarcely known a decade and a half ago. Few scientists could have predicted that these tools of basic research would come to dominate the field of food diagnostics. Today, a large number of cleverly designed assay formats using these technologies are available commercially for the detection in foods of practically all major established pathogens and toxins, as well as of many emerging pathogens. These tests range from very simple antibody-bound latex agglutination assays to very sophisticated DNA amplification methods. Although molecular biological assays are more specific, sensitive, and faster than conventional (often cultural) microbiological methods, the complexities of food matrices continue to offer unique challenges that may preclude the direct application of these molecular biological methods. Consequently, a short cultural enrichment period is still required for food samples prior to analysis with these assays. The greater detection sensitivity of molecular biological methods may also affect existing microbiological specifications for foods; this undoubtedly will have repercussions on the regulatory agencies, food manufacturers, and also consumers. PMID- 9219241 TI - Use of gas chromatography-ion trap tandem mass spectrometry for the detection and characterization of microorganisms in complex samples. AB - Gas chromatography-mass spectrometry (GC-MS) can be applied to detect and characterize microorganisms in clinical and environmental samples, and microbial contaminants in biotechnological production cultures. With this approach, unique microbial monomeric compounds, known as chemical markers, are used as analytes. In the present article, two GC-MS-based techniques, viz. GC-ion trap tandem MS (GC-MS-MS) and conventional quadrupole GC-MS used in the selected ion monitoring mode, were compared regarding their ability to detect 3-hydroxy fatty acids, muramic acid, and ergosterol (markers for endotoxin, peptidoglycan, and fungal biomass, respectively) in complex matrices. When using GC-MS-MS, daughter ion spectra were obtained for all markers present in amounts close to the detection limit of the GC-MS. Ion-trap GC-MS-MS shows great promise as a chemical marker analysis technique for application in clinical diagnosis, occupational and public health care, and biotechnology. PMID- 9219239 TI - Recent advances in transgenic technology. AB - Techniques that allow modification of the mammalian genome have made a considerable contribution to many areas of biological science. Despite these achievements, challenges remain in two principal areas of transgenic technology, namely gene regulation and efficient transgenic livestock production. Obtaining reliable and sophisticated expression that rivals that of endogenous genes is frequently problematic. Transgenic science has played an important part in increasing understanding of the complex processes that underlie gene regulation, and this in turn has assisted in the design of transgene constructs expressed in a tightly regulated and faithful manner. The production of transgenic livestock is an inefficient process compared to that of laboratory models, and the lack of totipotential embryonic stem (ES) cell lines in farm animal species hampers the development of this area of work. This article highlights recent progress in efficient trans gene expression systems, and the current efforts being made to find alternative means of generating transgenic livestock. PMID- 9219243 TI - Labeling of oligonucleotides with fluorescein. AB - The labeling of oligonucleotide probes using a fluorescein-labeled nucleotide is described. The reaction is characterized by careful control of the nucleotide and probe molar ratio in order to produce a tail that gives good detection sensitivity without compromising hybridization stringency control of the probe sequence. The labeling reaction can be easily monitored for incorporation of the fluorescent label and the probes can be used in many applications. PMID- 9219244 TI - Use of baculoviruses as biological insecticides. AB - Naturally occurring baculoviruses can be used to control a wide range of insect pests. Most baculoviruses are used as biopesticides, that is, they are sprayed onto high-density pest populations in a manner akin to the use of synthetic chemical pesticides. However, other strategies that use the biological features of the viruses are also possible and should increase as we expand our knowledge of baculovirus ecology. In order to develop a baculovirus control program, several areas need to be studied before progressing to large scale field studies and commercialization. These range from laboratory efficacy testing and the development of production systems to detailed study of pest behavior and the development of appropriate application strategies. PMID- 9219242 TI - Replication-competent retroviral vectors for expressing genes in avian cells in vitro and in vivo. AB - Replication-competent retroviral vectors based on Rous sarcoma virus (RSV) are becoming increasingly popular for expressing genes in both primary cell cultures and embryonic chick tissues in ovo. In this article, we review the features of RSV and its life cycle that make it suitable for use as a vector. We describe the design and use of the RCAS and RCAS (BP) series of vectors, which are currently the most widely used RSV-based vectors, illustrating both their strengths and weakness. Finally, we outline laboratory protocols suitable for the banding of these retroviral vectors. PMID- 9219245 TI - NMR spectroscopy of peptides and proteins. Practical considerations. AB - High resolution nuclear magnetic resonance (NMR) spectroscopy is the only method available for determining the three-dimensional structures of peptides and proteins in solution at atomic resolution. This article deals with a range of practical considerations associated with such studies, including sample preparation, instrumental setup, one- and two-dimensional NMR methods, interpretation of spectral data, and structure calculations. PMID- 9219247 TI - Chickens, cows, bulls, and bears. PMID- 9219246 TI - Efficient preparation of single-stranded DNA for in vitro selection. AB - In vitro selection of aptamers requires the reliable enzymatic preparation of large amounts of (+) single-stranded DNA molecules. This can be achieved by selective enzymatic digest of 5'-phosphorylated (-) strands from PCR products, a method already widely used in sequencing of PCR products. Here we present an adaptation of this method to prepare large pools of single-stranded DNA molecules for in vitro selection. PMID- 9219248 TI - Clinical investigators: the driving force behind drug discovery. PMID- 9219249 TI - Clinton's cloning ban may threaten genetic research. PMID- 9219250 TI - Sepsis therapy still shocking. PMID- 9219251 TI - Roche-Mannheim: cornering the diagnosis-led healthcare market. PMID- 9219252 TI - US bioethicists say continue human cloning moratorium. PMID- 9219254 TI - Biotech unruffled by setbacks. PMID- 9219253 TI - Europe/Japan face up to legal hurdles to cloning. PMID- 9219256 TI - Antibodies live long and prosper. PMID- 9219257 TI - Happy coupling: recruiting both antigen and effector function. PMID- 9219255 TI - Hemispherx: 300 patents, 20 staff. PMID- 9219258 TI - Accessories for naked DNA vaccines. PMID- 9219259 TI - Liposomal gene delivery: a complex package. PMID- 9219260 TI - Surface warfare against pathogens using mucosal vaccines. PMID- 9219261 TI - Entering the domain of neurotrophin binding. PMID- 9219262 TI - Molecular technologies for biodiversity evaluation: opportunities and challenges. PMID- 9219263 TI - Complement recruitment using bispecific diabodies. AB - We describe the engineering of antibody fragments produced in bacteria for recruitment of complement effector functions. From a phage display repertoire we isolated human antibody fragments directed against complement C1q, and linked these to lysozyme-specific antibody fragments, creating bispecific antibodies (diabodies). One diabody was able to recruit C1q, resulting in efficient lysis of lysozyme-coated sheep erythrocytes, and also induced rosette-formation of erythrocytes with human monocytes and phagocytosis after phorbol ester stimulation. These diabodies may have therapeutic applications requiring the activation of complement. PMID- 9219264 TI - Retargeting serum immunoglobulin with bispecific diabodies. AB - Monospecific antibody fragments produced in bacteria lack the Fc portion of antibodies, and are therefore unable to recruit natural effector functions. We describe the use of a bispecific antibody fragment (diabody) to recruit the whole spectrum of antibody effector functions by retargeting serum immunoglobulin (Ig). One arm of the diabody was directed against the target antigen, and the other against the serum Ig. The bispecific diabodies were able to recruit complement, induce mononuclear phagocyte respiratory burst and phagocytosis, and promote synergistic cytotoxicity towards colon carcinoma cells in conjunction with CD8+ T cells. Further, by virtue of binding to serum Ig their half-life (beta-phase) was increased fivefold compared to a control diabody of the same molecular weight. Such bispecific diabodies may provide an attractive alternative to monoclonal antibodies for serotherapy. PMID- 9219265 TI - Increasing the serum persistence of an IgG fragment by random mutagenesis. AB - The major histocompatibility complex (MHC) class I-related receptor FcRn is involved in regulating serum gammaglobulin (IgG) levels in mice. With the aim of increasing the serum half-life of a recombinant murine Fc gamma 1 fragment, the affinity for binding to FcRn at pH 6.0 has been increased by random mutagenesis of Thr252, Thr254, and Thr256 followed by selection using bacteriophage display. These residues were chosen as they are in proximity to the FcRn-IgG (Fc) interaction site. Two mutants with higher affinity (due to lower off-rates) than the wild-type Fc have been isolated and analyzed in pharmacokinetic studies in mice. The mutant with the highest affinity has a significantly longer serum half life than the wild type fragment, despite its lower off-rate from FcRn at pH 7.4. The results provide support for the involvement of FcRn in the homeostasis of serum IgGs in mice. The indications that a homologous FcRn regulates IgG levels in humans suggest that this approach has implications for increasing the serum persistence of therapeutic antibodies. PMID- 9219266 TI - Engineering of in vivo immune responses to DNA immunization via codelivery of costimulatory molecule genes. AB - Nucleic acid immunization is a novel vaccination technique to induce antigen specific immune responses. We have developed expression cassettes for cell surface markers CD80 and CD86, two functionally related costimulatory molecules that play an important role in the induction of T cell-mediated immune responses. Coimmunization of these expression plasmids, along with plasmid DNA encoding for HIV-1 antigens, did not result in any significant change in the humoral response; however, we observed a dramatic increase in cytotoxic T-lymphocyte (CTL) induction as well as T-helper cell proliferation after the coadministration of CD86 genes. In contrast, coimmunization with a CD80 expression cassette resulted in a minor, but positive increase in T-helper cell or CTL responses. This strategy may be of value for the generation of rationally designed vaccines and immune therapeutics. PMID- 9219267 TI - Improved DNA: liposome complexes for increased systemic delivery and gene expression. AB - To increase cationic liposome-mediated intravenous DNA delivery extruded DOTAP:cholesterol liposomes were used to form complexes with DNA, resulting in enhanced expression of the chloramphenicol acetyltransferase gene in most tissues examined. The DNA:liposome ratio, and mild sonication, heating, and extrusion steps used for liposome preparation were crucial for improved systemic delivery. Size fractionation studies showed that maximal gene expression was produced by a homogeneous population of DNA:liposome complexes between 200 to 450 nm in size. Cryo-electron microscopy examination demonstrates that the DNA:liposome complexes have a novel morphology, and that the DNA is condensed on the interior of invaginated liposomes between two lipid bilayers. This structure could account for the high efficiency of gene delivery in vivo and for the broad tissue distribution of the DNA:liposome complexes. Ligands can be placed on the outside of this structure to provide for targeted gene delivery. PMID- 9219268 TI - Oral vaccination of mice against tetanus with recombinant Lactococcus lactis. AB - To determine whether a protective immune response could be elicited by oral delivery of a recombinant bacterial vaccine, tetanus toxin fragment C (TTFC) was expressed constitutively in Lactococcus lactis and administered orally to C57 BL/6 mice. The antibody titers elicited were lower than those following intranasal immunization (a route already known to result in high-level systemic anti-TTFC immune responses) but the protective efficacy was the same order of magnitude. The serum antibody isotypes elicited were predominantly IgG1 and IgG2a. TTFC-specific fecal IgA responses could be detected following oral or intranasal immunization. Chemically killed lactococci administered via the intranasal route were also able to elicit serum antibody responses of similar levels and kinetics to those induced by live bacteria. PMID- 9219269 TI - Structure-based design and protein engineering of intersubunit disulfide bonds in gonadotropins. AB - Pairs of cystine residues were introduced in the alpha- and beta-subunits of human choriogonadotropin at positions with optimal geometries for the formation of disulfide bonds. Using the homology with luteinizing hormone and follicle stimulating hormone, similar mutations were carried out in these glycoprotein hormones. In nearly all mutants the corresponding disulfide bonds were formed leading to a non-natural, covalent linkage between the alpha- and beta-subunits. The mutants typically display wild-type receptor binding and bioactivity. The mutants with non-natural intersubunit disulfide bonds display enhanced thermostabilities relative to the corresponding heterodimeric glycoprotein hormones, rendering them candidates for long acting gonadotropins with enhanced shelf lives. PMID- 9219270 TI - Design of stable biologically active recombinant lutropin analogs. AB - Glycoprotein hormones are noncovalent heterodimers comprised of a common alpha subunit and a hormone-specific beta subunit. Secretion and biologic action of these hormones are dependent on the formation of the heterodimer. The human LH beta subunit is unique among the other beta subunits in that it assembles inefficiently with the alpha subunit. To bypass this rate-limiting step, we constructed the LH single chains where the carboxy terminus of beta was fused to the amino terminus of alpha subunit through a linker. Compared to the human LH heterodimer, the extent of secretion was greater for the tethers although the rate was dependent on the nature of the linker. The LH single chains were biologically active even though there was loss of recognition by a LH-specific monoclonal antibody. This suggests that receptor binding of the single chains is not impaired by changes in the heterodimeric configuration resulting from tethering the subunits. In addition, single chains exhibited a remarkably greater in vitro stability than the heterodimer, implying that these analogs will be useful as diagnostic reagents and that their purification will be facilitated. PMID- 9219271 TI - Immunoglobulin-like domains define the nerve growth factor binding site of the TrkA receptor. AB - Nerve growth factor (NGF) is involved in the development and maintenance of the nervous system. NGF binds with high affinity to the extracellular region of the tyrosine kinase receptor TrkA. This domain comprises leucine and cysteine rich motifs, followed by two immunoglobulin like (Ig-like) domains. We describe the expression and purification of recombinant Ig-like domains. Fluorescence and circular dichroism spectroscopy show that the protein is folded into a compact globular structure and contains mainly beta-sheet secondary structure. Recombinant protein binds to NGF and can inhibit NGF bioactivity both in vitro and in vivo. PMID- 9219272 TI - Regulation of the biological activity of glucagon-like peptide 2 in vivo by dipeptidyl peptidase IV. AB - Species-specific differences in the enzymatic inactivation of peptides is an important consideration in the evaluation of therapeutic efficacy. We demonstrate that glucagon-like peptide 2 (GLP-2), shown to be highly intestinotrophic in mice, promotes an increase in intestinal villus height but has no trophic effect on small bowel weight in rats. The reduced intestinotrophic activity of GLP-2 in rats is attributable to inactivation by the enzyme dipeptidyl peptidase IV (DPP IV). GLP-2(1-33) was degraded to GLP-2(3-33) following incubation with human placental DPP-IV or rat serum but not by serum from DPP-IV-deficient rats. Administration of rat GLP-2 to DPP-IV-deficient rats was associated with markedly increased bioactivity of rat GLP-2 resulting in a significant increase in small bowel weight. A synthetic GLP-2 analog, r[Gly2]GLP-2, with an alanine to glycine substitution at position 2, was resistant to cleavage by both DPP-IV and rat serum in vitro. Treatment of wild-type rats with r[Gly2]GLP-2 produced a statistically significant increase in small bowel mass. DPP-IV-mediated inactivation of GLP-2 is a critical determinant of the growth factor-like properties of GLP-2. PMID- 9219273 TI - Combining legal and business strategies to develop combinatorial chemistry businesses. PMID- 9219274 TI - Steps to building the virtual laboratory. PMID- 9219275 TI - Looking at thermocyclers. PMID- 9219276 TI - Critical issues in gene therapy commercialization. PMID- 9219277 TI - Prenatal diagnosis using fetal cells from the maternal circulation. AB - Current prenatal diagnosis relies on invasive methods such as amniocentesis and chorionic villus sampling. Because these methods carry a low, but finite risk of pregnancy loss, noninvasive genetic screening techniques are the focus of intense research. Isolating fetal cells from maternal blood for genetic analysis is the least invasive method currently being investigated. We discuss the various methods that have been used to isolate these cells. Nucleated red blood cells have emerged as the ideal fetal cell type. This is because they have the DNA material necessary for genetic analysis, they are consistently present in maternal blood, they can be easily identified based on their morphology, and they have a definite gestational life span. PMID- 9219278 TI - Peripartum pubic symphysis separation: a case series and review of the literature. AB - Peripartum pubic symphysis separation is a recognized complication of pregnancy with incidence estimates ranging from 1:300 to 1:30,000. Characteristic symptoms of symphyseal separation include suprapubic pain and tenderness with radiation to the back of legs, difficulty ambulating, and occasionally, bladder dysfunction. Clinical history, presenting symptoms, and response to therapy are sufficient to make the diagnosis, although radiographic documentation of symphyseal separation by x-ray or ultrasound are frequently used to confirm the diagnosis. The underlying etiology of symptomatic symphyseal separation has not been fully elucidated. Associations with multiparity, macrosomia, pathological joint loosening, and increased force placed on the pelvic ring have been suggested as possible etiologies. Conservative therapy, including bedrest, pelvic binders, ambulation devices, and mild analgesics usually result in a complete recovery with 4 to 6 weeks. The occurrence of a symphyseal separation should not significantly alter the management of subsequent pregnancies, and conservative therapy is recommended for any recurrence of symptoms. A retrospective review of our experience with 5121 deliveries from 1994 to 1995 found 9 cases of peripartum symphyseal separation, resulting in an incidence of 1 of 569 deliveries. Details regarding this case series and a review of the literature are presented. PMID- 9219279 TI - Prenatal ultrasonographic assessment of the middle cerebral artery: a review. AB - Published reports, case studies, and articles regarding ultrasonographic morphology, physiology, and pathophysiology of the fetal middle cerebral artery obtained from a MEDLINE search from 1966 through January 1997 were reviewed. Both transabdominal and transvaginal color Doppler ultrasonographic modalities may be used to assess fetal middle cerebral artery flow hemodynamics. Altered middle cerebral artery flow velocities may be noted in various medical conditions that include various behavioral states, term and preterm labor, maternal medications (anesthesia, tocolytics), fetal compromise (growth restriction and hypoxia), twin twin transfusion syndrome, invasive diagnostic procedures (amniocentesis and fetal blood sampling), fetal anemia and transfusion, in addition to intracranial fetal lesions (congenital anomalies and hemorrhage). In summary, knowledge of Doppler flow velocity of the fetal middle cerebral artery may assist prenatal diagnosis and management of complicated pregnancies. PMID- 9219280 TI - Low vision--optometry's opportunity. PMID- 9219281 TI - Colored filters and reading difficulties: a continuing controversy. PMID- 9219282 TI - National survey of the impact of low vision device use among veterans. AB - BACKGROUND: This report presents the results of a 2-year study of veterans' use of low vision devices (LVDs) which were prescribed and dispensed through he Blind Rehabilitation Centers (BRCs) and Visual Impairment Centers to Optimize Remaining Sight (VICTORS) of the Department of Veterans Affairs. METHODS: Two-hundred veterans using 740 LVDs were surveyed by telephone 12 to 24 months after the prescription/dispensing of the devices. Reliability (test-retest) and validity (content, criterion-related, and construct) were established for the survey. Primary analysis of the data was accomplished through tabular presentations. Factor analyses were used to describe prescription and use patterns. RESULTS: Most (85.4%) of the devices were still in use. Having a helper in the home was a demographic variable related to continued use. Neither age, acuity, nor etiology were related to continued use. Strong prescription and use patterns emerged. Most veterans reported receiving > 20 h of training and > 20 h of practice in the use of their LVDs. CONCLUSIONS: Most veterans who receive LVDs through the service delivery system of the Department of Veterans Affairs appear to use them for a wide variety of daily tasks and reported that they obtain a great deal of benefit from their use. PMID- 9219283 TI - Veterans' use of low vision devices for reading. AB - BACKGROUND: This report presents the results of a 2-year study of veterans' use of low vision devices (LVDs) which were prescribed and dispensed through the Blind Rehabilitation Centers (BRCs) and Visual Impairment Centers to Optimize Remaining Sight (VICTORS) of the Department of Veterans Affairs. METHODS: Two hundred veterans using 740 LVDs were surveyed by telephone 12 to 24 months after the prescription/dispensing of the devices. Reliability (test-retest) and validity (content, criterion-related, and construct) were established for the survey. Primary analysis of the data was accomplished through tabular presentations. Because most devices were used for reading, an exploratory data analysis was completed to further investigate successful use of LVDs for this task. Relationships of 21 variables with a definition of highly successful use, use and nonuse of LVDs for reading were evaluated. RESULTS: Only visual acuity provided a statistically significant predictor of use of LVDs for reading. LVDs in the lowest visual acuity grouping tend to be used either highly successfully, or fall into the nonuse category. The highly successful LVDs are primarily video magnifiers; the nonused LVDs tended to be spectacle magnifiers. CONCLUSIONS: This population is using devices extensively for reading, reporting frequencies of use of several times per day. PMID- 9219284 TI - Congenital nystagmus image motion: influence on visual acuity at different luminances. AB - In persons with congenital nystagmus (CN), the ability to integrate visual information over time can be limited by two factors--the duration of foveation periods and the temporal integration period of the visual system. The purpose of this study was to assess the relative importance of these two factors for visual acuity for targets of different luminances. We measured visual acuity using Landolt C targets at 5 luminance levels (50 to 0.005 cd/m2) in 6 observers with CN, and in 6 normal observers with comparable motion of the retinal image. To allow comparison, normal observers viewed the targets during image motion simulating jerk CN, with "foveation durations" ranging from 20 to 160 ms. In the normal observers, acuity improves as a function of the simulated foveation duration at all luminance levels. However, this improvement is larger and occurs at a faster rate at high than at low luminances. The more gradual improvement in acuity with simulated foveation durations at low luminances is consistent with a prolongation of the temporal integration period, which we estimate to range from approximately 140 to 380 ms over the 4 log unit range in luminance that we tested. In observers with CN, the change in acuity with luminance is similar, but not identical, to that in normal observers when the duration of the foveation periods is matched. We conclude that the integration of visual information may be limited by either the period of temporal integration or the duration of the foveation period in persons with CN, depending upon which is shorter at the luminance level under consideration. PMID- 9219285 TI - Preocular tear film characteristics of nonwearers and soft contact lens wearers. AB - The aims of the current investigation were to: (1) characterize (structure, volume, and stability) the preocular tear film of contact lens wearers and nonwearers and (2) test for any difference between contact lens wearers and nonwearers and between symptomatic and asymptomatic subjects. The tear film structure and stability were tested using the Tearscope in conjunction with the biomicroscope observation system. The tear prism height, which is indicative of the tear volume, was measured with the slitlamp. The study was carried out on 239 subjects (478 eyes) who attended our clinic for contact lens fitting. Of these, 184 were habitual daily soft contact lens wearers who had not been wearing contact lenses for at least 24 h; the other 55 were noncontact lens wearers. The results obtained showed that: (1) the stability of the tear film was correlated for two eyes of the same subject; (2) the structure, volume, and stability of the preocular tear film were similar for both groups; (3) no difference in tear film stability was found between asymptomatic and symptomatic contact lens wearers, but a significant difference was found between asymptomatic and symptomatic noncontact tact wearers; and (4) the stability of the tear film was influenced by the nature of the lipid layer present at the surface of the aqueous layer; the greatest stability was achieved when the lipid layer was thick and homogeneous (amorphous pattern). PMID- 9219286 TI - Size of cells collected from normal human subjects using contact lens cytology. AB - This paper describes how a soft contact lens can be used to harvest cells from the surface of the corneal epithelium. The procedure is called contact lens cytology (CLC). Cells were removed from a soft contact lens by irrigation and stained with acridine orange. Two methods for the measurement of cell size are described. First, cell size was measured using a computer-assisted technique, which calculated the area of the cell from its outline. The second method was simpler in that it required only a single measurement of the longest dimension of the cell (the cell length). To test the validity of this simpler method, cell area was compared with cell length in 185 cells. The resulting correlation (r = 0.92) suggests that the size of shed cells can be described adequately using cell length in place of the more time-consuming measurement of cell area. A mathematical relation can be used to convert cell length to cell area so that results from experimenters using different measures of size can be compared. When a large pool of cells collected by CLC was divided into four aliquots and cell length measured by two observers on two different days, there were no significant differences between observers or days. Thus, the technique does not depend on one observer, and it is unaffected by a 24-h delay in measurement. Cells were harvested from the corneal epithelium of normal human subjects. The number of cells collected from any single removal of the contact lens had a range of 10 to 175 cells, and a mean of 66.8 +/- 40.4 (N = 46). Cell length was measured and plotted as frequency histograms for both eyes of each subject. The range in cell length was from 10 to 80 microns. The mean cell length for individual subjects had a low of 26.5 +/- 9.0 microns and a high of 44.2 +/- 10.2 microns, with a grand mean for all right eyes of 36.0 +/- 5.1 microns, and a grand mean for all left eyes of 34.6 +/- 5.2 microns. The mean for all eyes was 35.3 +/- 5.1 microns. Composite histograms were created with the combined data from the 23 right eyes (N = 1310 cells), and the 23 left eyes (N = 1765 cells). Individual histograms and the composite histograms were not normally distributed. Peaks in the distributions suggest the presence of different subpopulations of cells, lending support to the hypothesis that there is more than one mechanism for cell shedding. PMID- 9219287 TI - Efficacy of hand washing procedures on bacterial contamination of hydrogel contact lenses. AB - The effect of various hand washing regimens on transfer of bacterial contaminants from the hands to a hydrogel contact lenses was evaluated. Each of 47 subjects performed 5 different hand washing procedures, and then handled a new, sterile hydrogel contact lens. The lenses were cultured to determine colony-forming units (CFUs) and microbial identity. Median CFUs on lenses handled after washing with water, soap and water, or soap and water followed by towel drying were higher than the median CFU for lenses handled after no hand washing. The median CFU for lenses handled after soap and water washing followed by an alcohol wipe was not different from the no washing group. The majority of the contaminants were identified as Staphylococcus epidermidis. These results show that ordinary hand washing alone does not decrease, and may even increase, the amount of contaminants transferred from the hands to a hydrogel lens. Use of an alcoholic wipe after hand washing reverses this effect. Hand washing is still recommended in contact lens hygiene for removal of more pathogenic contaminants. PMID- 9219288 TI - Prevalence of idiopathic corneal anomalies in a non contact lens-wearing population. AB - BACKGROUND: Successful clinic management of contact lens wear requires the differentiation of induced corneal changes from normal variation. Information concerning the type and prevalence of anomalous features occurring in the corneas of non contact lens wearers would assist the clinician to develop an accurate impression of the normal condition. METHODS: Seventy normal, asymptomatic, consecutively presenting, non contact lens wearers were subjected to an extensive biomicroscopic examination by a single clinician. RESULTS: Forty-nine percent of the sample were observed to have epithelial microcysts, 21% idiopathic scars, 20% endothelial bedewing, 10% epithelial vacuoles, 4% subepithelial microinfiltrates (SEMIs), and 4% Hudson-Stahli lines. Considerable day to day variation in the numbers of SEMIs was observed within subjects. CONCLUSION: These findings demonstrate that certain features which are frequently associated with contact lens wear occur idiopathically in the nonwearing eye. PMID- 9219289 TI - Irlen lenses do not improve accommodative accuracy at near. AB - The purpose of the experiment was to determine the influence of Irlen lenses on steady-state accommodation in successful users, as there is speculation that near accommodative dysfunction may be a factor in these patients. Monocular steady state accommodation in six successful Irlen patients was assessed for near blur stimuli (2, 3, and 4 D) either with nonfiltered spectacle refractive correction or their spectrally broad-band filtered spectacle refractive correction. Accommodation was measured subjectively using a Hartinger coincidence optometer. There was no significant difference in mean level of accommodation between the two conditions. Additionally, there was a small but significant increase in accommodative variability with the filtered prescription. The use of Irlen correction did not have any positive effect on monocular steady-state accommodation at near. PMID- 9219291 TI - Substance abuse versus substance dependence. PMID- 9219290 TI - Tonic vergence and vergence adaptation. AB - In the absence of an adequate visual stimulus, the eyes are typically converged by approximately 0.25 to 0.75 meter angles (MA). This vergence response (VR) was believed to reflect the level of tonic innervation to the extraocular muscles, and accordingly has been termed tonic vergence (TV). However, this estimation fails to consider the magnitude of the anatomical position of rest. The true typical value of TV is approximately 23 degrees. This paper will consider various aspects of this parameter, including both clinical and laboratory methods of measurement, and the relationship between TV and the distance heterophoria. In addition, the role of vergence (or prism) adaptation, i.e., the apparent change in TV after periods of sustained fixation, is discussed. This shift appears to result from the relatively prolonged decay of the slow fusional vergence response (VR), with no evidence for a change in the level of tonic innervation. On occasion, the decay of slow fusional vergence may take hours or even days to reach completion. This extended rate of decay will have a significant impact upon the clinical measurement of a number of binocular parameters, most notably the assessment of heterophoria under truly dissociated conditions (i.e., in the absence of any fusional VR). Furthermore, both the magnitude and rate of decay of vergence adaptation appear to vary with age, as well as the presence of oculomotor imbalance. It is concluded that the output of the slow fusional vergence mechanism, as reflected by the degree of vergence adaptation, makes a major contribution to the aggregate, sustained VR in most visually normal patients. PMID- 9219292 TI - Psychiatric research and the incompetent subject. PMID- 9219293 TI - Investigating the role of lipids in mood, aggression, and schizophrenia. PMID- 9219294 TI - The realities of clonazepam discontinuation. PMID- 9219295 TI - Setting behavioral health priorities: good news and crucial lessons from the Oregon Health Plan. PMID- 9219296 TI - Dual diagnosis treatment for patients with schizophrenia who are substance dependent. PMID- 9219297 TI - Assessing clinical outcomes: the community functioning of persons with serious mental illness. AB - OBJECTIVE: The demand to measure the clinical outcomes of persons with serious mental illness in the community is growing; however, there is no consensus about how to do this task. This paper identifies challenges in measuring the outcomes of persons with serious mental illness and reviews selected instruments that measure the community functioning of this population. METHODS: Papers in peer reviewed psychiatric journals for the years 1986 to 1996 were reviewed to select instruments that measure two or more domains of community functioning and for which data on reliability and validity have been published. Selected instruments were evaluated, focusing on their format, content, item scoring, length, and original sample population. RESULTS AND CONCLUSIONS: Challenges to measuring the community functioning of persons with serious mental illness include the multiplicity of domains that must be measured, the conflicting interests of various stakeholders involved in care, the limitations of self-report data, and other methodological problems. Nine instruments that met the study criteria were selected from the literature. Three are self-report instruments, and six are based on the report of an informant or independent rater. The instruments vary in length and in their original sample population. The content areas most consistently represented are self-care and social relationships. Life satisfaction, health status, psychiatric symptoms, and work skills are not consistently addressed. Individual instruments have additional limitations, including the absence of behavioral anchors for scale items and the lack of specificity to persons with serious mental illness. Effort should be directed toward sharing data across settings, measuring the effects of treatment interventions, and demonstrating the predictive validity of outcome data. PMID- 9219298 TI - Development of outcome indicators for monitoring the quality of public mental health care. AB - OBJECTIVE: The study attempted to develop a brief and integrated set of reliable and valid outcome measures that could be used by both consumers and providers to assess the quality of public mental health care. METHODS: A model of outcomes in four domains-consumer satisfaction, functioning, quality of life, and clinical status-was developed from the literature and from the priorities expressed by members of an advisory group of stakeholders. Based largely on extant measures, a consumer survey and a case manager survey were then created to assess these domains. A total of 236 adult consumers of mental health services from six community mental health centers in Washington State were surveyed. The four-item case manager survey to rate consumers' clinical status was completed by 163 of the participants' case managers. Scores and ratings on the survey were analyzed using correlational analysis and principal components analysis to determine whether the data provided empirical support for the four-domain model. RESULTS: Principal components analysis demonstrated support for the four-domain model. Internal consistency of the outcome indicators was adequate, and their concurrent validity was partly supported. CONCLUSIONS: The described outcome measures provide a practical, empirically supported structure for monitoring and improving public mental health services. PMID- 9219299 TI - Comprehensive case management in the private sector for patients with severe mental illness. AB - Intensive case management for severely psychiatrically ill patients is a relatively new phenomenon in the private sector. The authors describe a comprehensive case management program designed at Blue Cross Blue Shield of Massachusetts to meet the needs of the most severely ill psychiatric patients in a private managed care environment. The case management program emphasizes involvement of patients in creating comprehensive treatment plans; development of a relationship between case managers, patients and their families, and providers; and clinical coordination between the public and private sectors to create individualized treatment plans. The program's case managers are able to flex the benefit limitations of a managed care or indemnity plan to integrate public and private services and can enlist providers outside a managed care network. The paper describes service utilization by the first 33 patients who participated in the program for one year. PMID- 9219300 TI - The psychiatrist's role as medical director: task distributions and job satisfaction. AB - OBJECTIVES: Previous surveys of the alumni of Columbia University's fellowship in public psychiatry suggest that a large number of alumni fill positions as program medical directors. In contrast with agency medical directors, program medical directors work within team structures and maintain a high degree of clinical involvement. The fellowship faculty surveyed the alumni to catalog the tasks performed by program medical directors, agency medical directors, and staff psychiatrists and to determine the extent to which these tasks contribute to job satisfaction. METHODS: A survey form was developed using a list of tasks derived from the American Psychiatric Association's guidelines for psychiatrists working in organized mental health care delivery systems and from a recent article that surveyed job descriptions of psychiatrists in community mental health centers. The survey form was distributed to all current fellows and alumni in active practice (N = 89). RESULTS AND CONCLUSIONS: Seventy-two forms were returned, for a response rate of 81 percent. Respondents who were medical directors performed a greater variety of tasks and reported higher job satisfaction than those who were staff psychiatrists. Higher job satisfaction was related to a greater variety of tasks performed, especially tasks involving clinical collaboration. Most of the respondents were program medical directors rather than agency medical directors. The position of program medical director constitutes a relatively small and attainable step above that of staff psychiatrist. Agencies would do well to consider creating positions of program medical directors for their staff psychiatrists whenever feasible, and psychiatrists committed to public-sector careers should negotiate to have such positions. PMID- 9219301 TI - The impact of organizational factors on mental health professionals' involvement with families. AB - OBJECTIVE: Drawing on an organizational behavior framework, this study explored the impact of attitudinal, occupational, and organizational factors on mental health professionals' involvement with clients' families. METHODS: Data came from a survey conducted with psychiatric staff at the largest public hospital and the largest private hospital in Indianapolis between 1991 and 1993 as part of the Indianapolis network mental health study. Responses of 184 clinicians who provided direct care were analyzed using multiple regression to assess the impact of their attitudes toward families, job characteristics, and organizational work environment on the amount of contact they had with clients' families. RESULTS: Providers' attitudes toward families had no significant effect on the frequency of their contact with families. Job and organizational factors were the strongest predictors. Specifically, being a social worker or therapist and working on day and evening shifts were associated with increased involvement with families. Staff members' perceptions of how well their unit functioned were also positively correlated with frequency of contact with families. CONCLUSIONS: The organizational environment in mental health agencies has a significant influence on the extent to which professionals become involved with clients' families. Administrators and policy makers should give careful consideration to how the work environment encourages or limits mental health professionals' abilities and willingness to get more involved with clients' families. PMID- 9219302 TI - Effectiveness of treatment elements in a residential-work therapy program for veterans with severe substance abuse. AB - OBJECTIVE: This study evaluated outcomes of a residential-work therapy program for veterans with chronic, severe substance use disorders. METHODS: Admission and three-month outcome data were gathered for 496 veterans treated in the Department of Veterans Affairs' Compensated Work Therapy-Transitional Residence program. Multivariate techniques were used to assess the relationship between admission risk factors, treatment elements, and outcome measures. RESULTS: Substantial improvement was observed in substance abuse and most other outcome domains, with 65 percent of the sample reporting no substance use during the three months after discharge. The most powerful baseline prognostic factors were functional status and frequency of social contact. Of 14 significant relationships observed between the intensity of treatment elements and outcomes, 12 were in the expected direction, associating more intensive treatment with improvement. The largest number of significant relationships with outcome were observed for weekly toxicology screens, earnings, and length of stay. CONCLUSIONS: The study results support the effectiveness of a rehabilitative approach to the treatment of severe substance abuse that combines residential support with demands for responsible behavior. PMID- 9219304 TI - The moderating effects of race on return visits to the psychiatric emergency room. AB - OBJECTIVE: Racial differences in variables that predict return to the psychiatric emergency room were examined. METHODS: A random sample of 319 clients was obtained from the logs of a psychiatric emergency room of a state-operated, acute care psychiatric hospital. The dependent variable was a return visit to the psychiatric emergency room within 18 months of the index visit. Separate logistic regression equations were calculated for African Americans (N = 163) and Caucasians (N = 156) to estimate the moderating effects of race. RESULTS: Four variables predicted return to the emergency room for both African Americans and Caucasians: previous visits to the psychiatric emergency room, previous psychiatric hospitalizations, current receipt of outpatient treatment, and nonreceipt of aftercare following the index visit to the emergency room. Three unique predictors were found for African Americans: never having been married, not living in stable housing, and not being admitted at the index visit. CONCLUSIONS: Generally, repeat visitors from both racial groups tended to be chronic users of psychiatric services who may be using the psychiatric emergency room for routine psychiatric care. However, race was also an important moderator variable; several risk factors predicted a return visit only for African Americans. PMID- 9219303 TI - Housing status among formerly homeless dually diagnosed adults. AB - OBJECTIVE: Residential outcomes of homeless adults with severe mental illness and a substance use disorder were studied over 18 months during which participants received integrated dual diagnosis services and housing supports based on a continuum model. METHODS: Interviews with 158 participants at baseline and at six , 12-, and 18-month follow-ups assessed housing status, residential history, substance abuse and progress toward recovery, psychiatric symptoms, and quality of life. Complete data were available for 122 participants. If participants lived continuously in high-quality housing with no housing loss or nights of homelessness during the final six months of the study, they were classified as having stable housing. RESULTS: Of the 122 participants for whom complete data were available, 64 (52 percent) achieved stable housing. Most participants who achieved stable housing first entered staffed and supervised housing and then moved to independent arrangements by the end of the study. Stable housing during the final evaluation period was associated with lower substance use, greater progress toward substance abuse recovery, and higher quality of life. Final housing status was not predicted by baseline variables but was predicted by progress toward recovery during months 0 to 6 and 6 to 12 and by less severe drug use during months 6 to 12. Participants who abused no illicit drugs during months 6 to 12 were almost three times as likely to achieve stable housing as those who abused illicit drugs. CONCLUSIONS: Housing stability is strongly mediated by substance abuse and progress toward recovery. Nevertheless, when formerly homeless persons with dual diagnoses are provided integrated dual diagnosis treatment, they can gradually achieve stable housing. PMID- 9219305 TI - A survey of state and VA policies on psychiatrists as primary care providers. AB - Fifty-five state and territorial commissioners of mental health and chiefs of psychiatry at 158 Department of Veterans Affairs medical centers were surveyed about current policies related to psychiatrists' roles as primary care providers in state and VA facilities. About half the respondents indicated that psychiatrists provided primary medical care or indicated interest in having psychiatrists provide such care. Less than half of this group limited such care to specific patient populations, and less than 25 percent required specific training for providers. The survey results indicate that opportunities for psychiatrists to provide primary care exist in many state and VA facilities but that no generally accepted guidelines or training standards for such practice have been developed. PMID- 9219306 TI - Dependence on public financial support early in the course of schizophrenia. AB - Although most patients with schizophrenia rely on public financial support, little is known about how soon after the onset of illness such dependence occurs. Forty-eight patients with schizophrenia were followed for a mean of five years after their first hospitalization to examine their reliance on public support. At one year after their first hospitalization, 27 subjects (56 percent) were primarily supported by social service agencies. Once such support was initiated, it was maintained throughout the entire follow-up period for all patients except two. The findings indicate that dependence on public financial assistance begins very early in the course of illness for most hospitalized patients with schizophrenia. PMID- 9219307 TI - A county survey of mental health services in drug treatment programs. AB - Forty-five administrators of drug treatment programs in Los Angeles County were surveyed about the adequacy of mental health services within their program and the drug treatment system. Approximately half agreed that dually diagnosed clients are not served within the system, and the majority noted that their programs restrict admission of such clients. Administrators of outpatient drug free programs and methadone maintenance programs were more likely to characterize their mental health services as inadequate or unavailable than were administrators of other types of programs. The findings suggest the need to increase awareness of the treatment needs of dually diagnosed clients in drug treatment programs. PMID- 9219308 TI - Children are left out. AB - The author suggests that large numbers of children and adolescents in America are being left out of mental health and mental health care. He discusses areas, such as poverty, teenage pregnancy, and violence, in which public policy has failed. He calls for interdisciplinary collaboration in work with children and emphasizes the need for psychiatrists and allied health professionals to take major public health and government policy roles to ensure that child and adolescent health and mental health are taken seriously. PMID- 9219309 TI - Smoking and response to psychotropic drugs. PMID- 9219310 TI - Recidivism in Slovenia. PMID- 9219311 TI - Diversity in VA settings. PMID- 9219312 TI - Mental health parity. PMID- 9219313 TI - Mechanism of action of aspirin-like drugs. AB - Nonsteroid antiinflammatory drugs (NSAIDs) or aspirin-like drugs act by inhibiting the activity of the cyclooxygenase (COX) enzyme. Two isoforms of COX exist, COX-1, which is constitutively expressed, and COX-2, which is an inducible isoform. Prostaglandins synthesized by the constitutively expressed COX-1 are implicated in the maintenance of normal physiological function and have a 'cytoprotective' action in the stomach. COX-2 expression is normally low but is induced by inflammatory stimuli and cytokines. It is thought that the antiinflammatory actions of NSAIDs are caused by the inhibition of COX-2, whereas the unwanted side effects, such as gastrointestinal and renal toxicity, are caused by the inhibition of the constitutively expressed COX-1. Individual NSAIDs show different selectivities against the COX-1 and COX-2 isoforms. NSAIDs that are selective towards COX-2, such as meloxicam, may have an improved side-effect profile over current NSAIDs. In addition to their use as antiinflammatory agents in the treatment of rheumatoid arthritis and osteoarthritis, selective COX-2 inhibitors may also be beneficial in inhibiting colorectal tumor cell growth and in delaying premature labor. PMID- 9219315 TI - Nonsteroidal antiinflammatory drugs, ulcers and risk: a collaborative meta analysis. AB - Nonsteroidal antiinflammatory drugs (NSAIDs) are associated with a high prevalence of gastrointestinal toxicity. Comparisons of the risks associated with individual NSAIDs are needed, but most clinical studies in this area are not ideally suited for use in meta-analyses. These difficulties can be overcome by using strict criteria for the inclusion of studies, and by reanalyzing previous data, updated by authors where possible. In doing so, this meta-analysis compared the relative risk of serious toxicity associated with 14 NSAIDs from 12 studies with the risks of ibuprofen. These results were supported by a novel "summary ranking" analysis, which was weighted to limit the influence of smaller studies. Wide variations in relative risk among NSAIDs were observed with piroxicam and azapropazone being the most toxic. Ibuprofen was associated with the least risk, probably because of its widespread use as a low-dose analgesic. Five studies provided comparative data on NSAIDs at "high" and "low" doses (as defined in the original reports), showing that the risk of toxicity was dose related. Furthermore, at full antiinflammatory doses, the risk associated with ibuprofen was similar to that of naproxen and diclofenac. These analyses show that NSAIDs vary more in toxicity than in efficacy. First-line therapy should be started with the lowest effective dose of a less toxic NSAID, moving to higher doses or a more toxic NSAID only if the clinical situation demands it. Newer NSAIDs, such as selective cyclooxygenase-2 inhibitors, may provide safer antiinflammatory therapy. PMID- 9219314 TI - The gastroenterologist's caseload: contribution of the rheumatologist. AB - Rheumatological conditions often give rise to gastrointestinal (GI) symptoms and vice versa, but the greatest point of contact between rheumatologists and gastroenterologists arises through gastrointestinal toxicity resulting from the use of nonsteroidal antiinflammatory drugs (NSAIDs). Standard NSAIDs are toxic to the entire GI tract. The point prevalence of ulcers in patients on long-term NSAID treatment is about 20% and the annual incidence of complications in these patients is 1% to 4%; 1,200 patients in the United Kingdom die each year as a result. Withdrawing NSAID treatment, or reducing the dose, is not always possible, and approximately 25% of patients require continued long-term antiinflammatory treatment. Misoprostol and acid-suppressing drugs such as ranitidine and omeprazole are helpful as prophylactic treatment in high risk patients, and in healing established ulcers, especially in patients carrying Helicobacter pylori, but there is a pressing need for new, safer antiinflammatory drugs toease the burden of NSAID gastropathy. Selective inhibition of the COX-2 isoform of cyclooxygenase has been proposed as a new and safer therapeutic option. Clinical studies with meloxican, a selective COX-2 inhibitor, support this view. Meloxicam has been extensively studied in arthritis patients and combines antiinflammatory efficacy with a lower incidence of GI toxicity than currently available NSAIDs. PMID- 9219316 TI - Meloxicam: selective COX-2 inhibition in clinical practice. AB - Nonsteroidal antiinflammatory drugs (NSAIDs) exert their actions by inhibiting cyclooxygenase (COX). It has recently been postulated that NSAIDs' antiinflammatory efficacy arises from inhibition of the COX-2 isoform of cyclooxygenase, whereas inhibition of the COX-1 isoform produces the troublesome and sometimes serious gastric and renal side effects of NSAIDs. A relatively selective COX-2 inhibitor, such as meloxicam, may combine antiinflammatory efficacy with improved tolerability. In volunteers, indomethacin 75 mg, but not meloxicam 7.5 mg, inhibited renal prostaglandin E2 excretion and platelet aggregation (COX-1 mediated effects). Double-blind, randomized trials in osteoarthritis and rheumatoid arthritis patients have shown equivalent antiinflammatory efficacy among meloxicam 7.5 mg or 15 mg and diclofenac 100 mg, naproxen 750 mg, and piroxicam 20 mg. In a double-blind, placebo-controlled trial, meloxicam (7.5 or 15 mg) caused less endoscopically detected gastrointestinal (GI) damage (Lanza scale) than piroxicam 20 mg. The MELISSA study, a double-blind, randomized, 28-day trial in over 9,000 patients showed that meloxicam 7.5 mg caused statistically less total GI toxicity, dyspepsia, abdominal pain, nausea and vomiting, and diarrhea than diclofenac 100 mg, despite equivalent reductions in pain on movement for each treatment. A global safety analysis of clinical trials, representing over 5,600 patients and comprising 170 and 1,100 patient-years of exposure for meloxicam 7.5 mg and 15 mg, respectively, showed that meloxicam caused less GI toxicity and fewer peptic ulcers and GI bleeds than naproxen, diclofenac, or piroxicam. The renal safety profile and incidence of liver function abnormalities with meloxicam is equivalent to other NSAIDs available for clinical use. In conclusion, relatively selective COX-2 inhibition exemplified by meloxicam may offer effective symptom relief with an improved GI tolerability profile. PMID- 9219317 TI - Differentiating among nonsteroidal antiinflammatory drugs by pharmacokinetic and pharmacodynamic profiles. AB - Cyclooxygenase (COX)-1 inhibition by nonsteroidal antiinflammatory drugs (NSAIDs) is associated with gastrointestinal and renal toxicity, while COX-2 inhibition has beneficial antiinflammatory, analgesic, and antipyretic effects. The measurement of inhibitory potency of NSAIDs on COX isoforms is strongly influenced by variations in experimental conditions in different models. The concentration of protein in the assay medium is particularly important because NSAIDs are highly protein-bound in vivo, and assays based on human whole blood provide the most suitable test system available. Of the drugs investigated in the human whole blood assay, only meloxicam showed selectivity for COX-2 at therapeutically relevant concentrations. In contrast, standard NSAIDs, such as naproxen, inhibited both COX isoforms to an equal degree at therapeutically relevant concentrations. Thus, the selectivity of NSAIDs against COX-2 relative to COX-1 may provide a means of differentiating between them according to their risk of toxicity at therapeutic doses. This may be more clinically meaningful than the traditional classification of NSAIDs based on chemical structure. PMID- 9219319 TI - Biology and medicine of sailing. An update. AB - The sport scientist's understanding of the biomechanics and physiology of sailing, together with its application to nutrition, training and injury prevention in the elite competitor, has continued to develop over the past decade. Very large mechanical forces are imposed in the vertical axis of the body, which give rise to frequent complaints to low back and knee pain and, occasionally, even to muscle rupture. Training programmes should emphasise the development of isometric endurance in the relevant muscle groups, such preparation continuing throughout the winter months. The oxygen cost of sailing is relatively light, and development of aerobic fitness should be advocated for reasons of general health rather than competitive success. Because of the intense muscle contractions that are developed during competition, heart rates and blood pressures are high in relation to oxygen consumption. However, during normal sailing, tacking and fluctuations of wind speed limit the development of muscle fatigue. In contrast to the operation of small craft, the crew of large ocean going vessels may have a very high daily energy expenditure, probably related to difficulty in relaxing at any point of day or night. Windsurfers face similar physiological demands to the dinghy sailor and they also have frequent complaints of back pain. Knowledge of the relevant health issues remains limited, even among elite competitors, and there remains substantial scope for increased education of team members. PMID- 9219318 TI - Effects of exercise on chromium levels. Is supplementation required? AB - It is estimated that most individuals are not ingesting sufficient amounts of chromium in their diets. Although there is little information on chromium intake in athletes, many athletes ingest more calories than do non-athletes so their chromium intake should be adequate. However, athletes who restrict calories to maintain low bodyweights could compromise their chromium status. Some evidence also shows that exercise may increase chromium loss into the urine. At present, it is not known whether this loss necessitates additional chromium in the diet or whether the body will increase retention in response to the loss. Chromium deficiency is thought to contribute to glucose intolerance and unhealthy blood lipid profiles. The primary function of chromium is to potentiate the effects of insulin, and thereby alter glucose, amino acid and fat metabolism. Chromium supplements have been purported to increase muscle mass and decrease body fat. However, the preponderance of evidence has not supported this claim. There is little information available on the long term use of chromium supplements, but at present, supplements within the Estimated Safe and Adequate Daily Dietary Allowance (ESADDI) level do not appear harmful. The prudent course of action for athletes would be to ingest foods rich in chromium and perhaps take a multivitamin/mineral supplement containing no more than the ESADDI of chromium. PMID- 9219320 TI - The effect of endurance training on reproductive function in male runners. A 'volume threshold' hypothesis. AB - Investigations on reproductive function in male athletes are not as abundant in the literature as the research available on female athletes. The primary reason for this is the absence of an obvious clinical sign indicative of an alteration in reproductive function in male athletes. While alterations in the reproductive status of female athletes may be easily detected by the loss of menstrual regularity, a distinctive clinical sign reflective of reproductive dysfunction in the male is not apparent. In male runners, an effect of endurance training on reproductive function related to a specific 'volume threshold' of training is proposed. Data are supportive of this 'volume threshold' effect, provided careful and consistent definitions of volume of training are applied. In fact, if volume of training is carefully defined endurance-trained male runners exhibit a rather consistent range of subclinical modifications in the gonadal hormones and semen profile, and clinical (oligospermia) alterations in reproductive function. The precise mechanism responsible for these observed alterations remains unknown, although several peripheral and central mechanisms have been suggested. Clearly, more research is necessary to confirm, and to elucidate, the nature of the 'volume threshold' hypothesis in male runners. PMID- 9219321 TI - Anthropometric and physiological characteristics of rugby union football players. AB - Rugby union enjoys worldwide popularity, but there is a lack of comprehensive research into the anthropometric and physiological characteristics of its players and the demands of the game, particularly at the elite level. One of the possible explanations for this is that the sport has previously been primarily concerned with the aspects of skill related to the game, rather than the physical and physiological requirements. However, with the increased physiological demands being placed on the elite players (using the British Isles as an example), with the recent introduction of professionalism, regional championships, the World Cup and major tours, information about the demands of the game and the assessment of, and methods of improving, the anthropometric and physiological characteristics of its players, are of paramount importance. Match analysis has indicated that rugby is an interval or intermittent sport and players must be able to perform a large number of intensive efforts of 5 to 15 seconds' duration with less than 40 seconds' recovery between each bout of high intensity activity. These observations, together with the metabolic responses during the game, give some insight into its physiological demands and are a prerequisite in the development and prescription of training programmes by coaches in preparing individual players for competition. The results from studies reporting the anthropometric and physiological characteristics of rugby union players observed that these individuals had unique anthropometric and physiological attributes which depended on positional role and the playing standard. These have important implications for team selection and highlight the necessity for individualised training programmes and fitness attainment targets. PMID- 9219322 TI - Hamstring injuries. Current trends in treatment and prevention. AB - Pre-exercise stretching and adequate warm-up are important in the prevention of hamstring injuries. A previous mild injury or fatigue may increase the risk of injury. Hamstring muscle tear is typically partial and takes place during eccentric exercise when the muscle develops tension while lengthening, but variation in injury mechanisms is possible. Diagnosis of typical hamstring muscle injury is usually based on typical injury mechanism and clinical findings of local pain and loss of function. Diagnosis of avulsion in the ischial tuberosity, with the need for longer immobilisation, and a complete rupture of the hamstring origin, in which immediate operative treatment is necessary, poses a challenge to the treating physician. X-rays, ultrasonography or magnetic resonance imaging (MRI) may be helpful in differential diagnostics. After first aid with rest, compression, cold and elevation, the treatment of hamstring muscle injury must be tailored to the grade of injury. Conservative treatment is based on a knowledge of the biological background of the healing process of the muscle. Experimental studies have shown that a short period of immobilisation is needed to accelerate formation of the granulation tissue matrix following injury. The length of the immobilisation is, however, dependent on the grade of injury and should be optimised so that the scar can bear the pulling forces operating on it without re rupture. Mobilisation, on the other hand, is required in order to regain the original strength of the muscle and to achieve good final results in resorption of the connective tissue scar and re-capillarisation of the damaged area. Another important aim of mobilisation--especially in sports medical practice--is to avoid muscle atrophy and loss of strength and extensibility, which rapidly result from prolonged immobilisation. Complete ruptures with loss of function should be operated on, as should cases resistant to conservative therapy in which, in the late phase of repair, the scar and adhesions prevent the normal function of the hamstring muscle. PMID- 9219323 TI - Coagulation activation in patients undergoing directional coronary atherectomy. AB - Restenosis is a major problem of percutaneous transluminal coronary angioplasty (PTCA) and related procedures. To better understand the underlying pathophysiologic mechanisms, coagulation and fibrinolytic variables were analysed prospectively in 35 patients after directional coronary atherectomy (DCA) and in 20 control patients undergoing diagnostic heart catheterisation and coronary angiography. Blood samples were taken before and 1 h, 24 h and 48 h after the procedure. No subacute thrombosis or unstable angina were documented in any patient. In 8 out of these 35 patients late restenosis was diagnosed during follow-up angiography 3-6 months after DCA. In these 8 patients prothrombin fragments (F1 + 2) rose from 0.7 to 0.9 nmol/l (P < 0.01) and thrombin antithrombin III complexes (TAT) from 2.9 to 6.0 micrograms/l (P < 0.01), but not significantly in 27 patients without restenosis and in the control patients. In patients with late restenosis plasminogen activator inhibitor (PAI-1) also increased from 2.4 to 4.9 U/ml (P < 0.05) 24 h after DCA while there were no significant changes in patients without restenosis and in control patients. D Dimer/TAT ratio reflecting the balance between clotting activation and fibrinolysis was significantly lower after 24 h in restenosis patients. The findings suggest that coagulation activation and hypofibrinolysis during 48 h after DCA might be associated with the development of late restenosis. PMID- 9219324 TI - Effect of bezafibrate on hypercoagulability assessed by fluorogenic prothrombin time in hyperlipidemic patients with non-insulin-dependent diabetes mellitus. AB - We investigated the usefulness of the fluorogenic prothrombin time (FPT) for detection of hypercoagulability and its association with hyperlipidemia in 19 patients with non-insulin-dependent diabetes mellitus (NIDDM) and 10 healthy control subjects, compared with plasma levels of fibrinogen, plasminogen activator inhibitor-1 (PAI-1) and D-dimer. We also evaluated the effect of bezafibrate on hypercoagulability in 10 hyperlipidemic NIDDM patients. The plasma levels of FPT and fibrinogen were significantly higher in hyperlipidemic NIDDM patients than in normolipidemic NIDDM patients and controls. Plasma levels of PAI 1 and D-dimer were significantly higher in normolipidemic and hyperlipidemic NIDDM patients compared with controls. The FPT was correlated with the HbA1c, the body mass index, and levels of total cholesterol, fibrinogen and PAI-1. Six months therapy with bezafibrate reduced the levels of FPT, triglycerides and basal insulin, but did not alter levels of fibrinogen, PAI-1 and D-dimer. Our results showed that the FPT was useful for detection of hypercoagulability and evaluation of the effect of drugs. The increased FPT in patients with NIDDM suggested that hypercoagulability was present in association with hyperlipidemia. PMID- 9219326 TI - An improved test to identify aPC-resistant factor V-Leiden. AB - A functional clotting assay was recently reported to detect the factor V-Leiden mutation (R506Q) in patients receiving oral anticoagulants and in patients with Lupus Anticoagulant inhibitor. The original assay (Dahlback) to detect resistance to activated protein C (aPC-resistance) frequently gave unreliable findings in these patients. The change in the method is the use of bovine thromboplastin in a PT-derived assay, with a 1:10 sample dilution. In these conditions a reference normal plasma gives a prolongation of more than 35 seconds, while samples with a heterozygous or homozygous mutation give a prolongation respectively of 10-18 seconds or less 2 seconds. The test is easily automated and quickly performed on ACL/3000, and the reproducibility is good. PMID- 9219325 TI - Antithrombotic effects of NE-6505, a novel anion-binding exosite inhibitor. AB - NF-6505, a bi-O-Tyr-sulfated decapeptide, which specifically interacts with the anion-binding exosite of the thrombin molecule, was chemically synthesized and assessed for its antithrombotic effects in vitro and in vivo. The IC50 value of this peptide on fibrin-clot formation in vitro was about 0.05 microgram/ml, which indicated a potency similar to that of a recombinant hirudin. NF-6505 caused a 2 fold prolongation of activated partial thromboplastin time when intravenously administered at 1 mg/kg in rats. In a rat venous thrombosis model, a bolus intravenous administration of this peptide dose-dependently inhibited the thrombus formation with an ED50 value of 0.03 mg/kg, a value smaller than that of recombinant hirudin (ED50 = 0.1 mg/kg) or of argatroban (ED50 = 0.2 mg/kg). These results suggest that NF-6505 is a highly potent and safe agent for the clinical treatment of venous thrombosis diseases. PMID- 9219327 TI - Binding of the von Willebrand factor A1 domain to histone. AB - Activation of the von Willebrand Factor (vWF) A1 domain is a critical factor in regulating the interaction of vWF with its platelet membrane receptor, the glycoprotein (GP) Ib-IX-V complex. This activation controls vWF-dependent platelet adhesion at high shear. The vWF-GP Ib-IX-V interaction is induced in vivo by exposure of platelet-rich plasma to high shear force, or by association of vWF with one or more unidentified components of the subendothelial matrix. In vitro, soluble vWF is activated to bind to platelets by nonphysiological modulators, such as the bacterial glycopeptide, ristocetin, or the snake venom protein, botrocetin, or by removal of negatively-charged sialic acid residues. Analysis of vWF modulators and the very marked charge asymmetry of amino acid sequences within the A1 domain has led to an electrostatic model for vWF modulation. Endothelial membrane/matrix and detergent-soluble fractions of human placenta were screened for the ability to bind vWF by electrophoresis of extracts on SDS-polyacrylamide gels, electrotransferring to nitrocellulose and probing with fluid-phase 125I-labeled vWF or a 39/34-kDa vWF fragment (Leu-480-Gly-718) that encompasses the A1 domain. In the course of these studies, it was found that both vWF and the 39/34-kDa vWF fragment bound strongly to histone. Purified soluble histone also bound vWF since, like ristocetin, it induced vWF flocculation. Histone binding to vWF did not activate or inhibit vWF binding to platelets. While the vWF-histone interaction has no conceivable physiological role, it suggests that binding to the A1 domain of vWF alone is insufficient to modulate vWF adhesive activity. This implies that specific interactions of the vWF A1 domain with either ristocetin or botrocetin are required for GP Ib-IX-V recognition to occur. PMID- 9219328 TI - Vasomodulatory action of clopidogrel and ticlopidine. AB - Clopidogrel is a thienopyridine derived antiplatelet drug that has currently undergone extensive clinical trials in the management of various arterial disorders related to platelet activation. While the proposed mechanism of its pharmacologic action is believed to be the inhibition of ADP mediated direct and indirect actions on platelet adhesion/aggregation and other activation processes, several other observed pharmacologic actions suggest that this drug may also have additional sites of action. Ticlopidine also belongs to the same class of ADP receptor inhibitor and is extensively used for prevention of ischemic disorders. In order to investigate the vasomodulatory action of clopidogrel and ticlopidine, rabbit and rat isolated tissue preparation systems were used. Clopidogrel and ticlopidine were found to produce dose dependent vasomodulatory actions in rabbit or rat treated with 30 minutes intravenous administration. The aortas harvested from both the rabbits and rats treated with clopidogrel or ticlopidine exhibited marked desensitization to the serotonin, endothelin-1, serum and platelet rich plasma/arachidonic acid mixtures. Both control rabbit aortic rings and rat aortic strips did not produce any inhibition of the serotonin induced contraction. These data suggest that clopidogrel and ticlopidine plays an important role in producing these vasomodulatory actions. Furthermore these observations indicate that both the clopidogrel and ticlopidine also modulate the vascular sites which may be contributory to the observed clinical effects. PMID- 9219329 TI - Measurement of factor VII and of activated factor VII in healthy individuals and in prothrombin complex concentrates. AB - We established reference ranges for factor VII clotting activity (FVII:C), factor VII amidolytic activity (FVII:AM), and activated factor VII (FVIIa) in 102 healthy individuals. The reference ranges were 65-160 U/100 ml, 70-165 U/100 ml, and 30-170 mU/ml, respectively (2.5 and 97.5 percentiles). Freezing and thawing of the plasma samples had no influence on the assay results. Due to the small sample size, the results were not influenced by gender, age, smoking habits, and oral contraceptive use. The plasma levels of FVII:C, FVII:AM, and FVIIa were significantly correlated with each other. The significant correlation between FVIIa and FVII:AM indicates that FVIIa is not completely independent of circulating FVII mass. There was also a significant, though weak, correlation between FVIIa and FVII:C/FVII:AM ratios. Sixteen batches of prothrombin complex concentrates (PCC) from 3 manufacturers were also analysed. FVIIa could be detected in all preparations, with considerable variations from batch to batch. In contrast to the results obtained in plasma from normal individuals, there was a close correlation between FVIIa and FVII:C/FVII:AM ratios. The preparations could be characterized by their FVII and FVIIa potencies and by their FVII:C/FVII:AM ratios. In PCC, FVII:C was very strongly correlated with FVIIa, whereas no significant correlation was observed between FVII:AM and FVII:C and between FVII:AM and FVIIa, respectively. These results demonstrate that the FVII:C assay used is sensitive for detecting FVIIa. Thus, we cannot confirm that FVIIa sensitivity of one-stage clotting assays for FVII:C is low when a rabbit thromboplastin and a non-adsorbed FVII-deficient plasma is used. PMID- 9219330 TI - Impaired clot lysis by rt-PA catalyzed mini-plasminogen activation. AB - The fibrinolytic system contains a proenzyme plasminogen (Plg) which is converted to plasmin (Plm) by the action of Plg activators. Physiological Plg activators are: tissue-type plasminogen activator (t-PA) and urokinase-type plasminogen activator. Plg was shown to be further cleaved by leukocyte elastase producing several fragments, one of which is called mini-plasminogen (mini-Plg) or neo plasminogen Val442. In this paper we studied whether mini-Plg is able to produce clot lysis when it is activated by rt-PA in purified systems and in Plg depleted normal plasma. We found that mini-Plg clot lysis time was longer than that of Plg. Clot lysis times were 2.3 minutes +/- 0.06 for Plg and 9.8 minutes +/- 0.1 for mini-Plg. Mini-Plg is less efficient than Plg in producing clot lysis at all studied concentrations (0.1-1.2 microM). In Plg depleted normal human plasma mini Plg is unable to produce complete clot lysis in presence of rt-PA. Although mini Plg can be activated to mini-Plm by rt-PA, these results show that the activation process is insufficient to produce an efficient clot lysis. PMID- 9219331 TI - Effect of erythrocytes and prostacyclin production in the effect of fructose and sorbitol on platelet activation in human whole blood in vitro. AB - We analyzed the in vitro effects of sorbitol and fructose on platelet function. Sorbitol and fructose increased platelet aggregation induced with adenosine diphosphate (ADP) or collagen in whole blood, but had no effect in platelet-rich plasma. The concentration that increased basal aggregation by 50% with ADP as the inducer was 12.89 +/- 1.55 mmol/L for fructose, and 18.99 +/- 2.01 mmol/L for sorbitol. When collagen was the inducer, these concentrations were 15.02 +/- 0.98 mmol/L for fructose, and 12.94 +/- 1.57 mmol/L for sorbitol. Both sugars increased, in a concentration-dependent way, the proaggregatory effect of erythrocytes, and erythrocyte uptake of adenosine. Time to uptake of 50% adenosine was 2.1 +/- 0.3 min in control samples, 0.14 +/- 0.01 min in the presence of fructose, and 0.23 +/- 0.03 min with sorbitol. Both sugars reduced vascular prostacyclin synthesis, with 50% inhibitory concentrations of 26.48 +/- 1.97 mmol/L for fructose, and 39.53 +/- 2.81 mmol/L for sorbitol. Both sugars also increased arterial lipid peroxidation by 30% (sorbitol) and 23% (fructose). We conclude that these two sugars enhance platelet function and disrupt the thromboxane/prostacyclin ratio. PMID- 9219332 TI - Comparison of two ELISAs for platelet factor 4. PMID- 9219333 TI - BTN1, a yeast gene corresponding to the human gene responsible for Batten's disease, is not essential for viability, mitochondrial function, or degradation of mitochondrial ATP synthase. AB - The Saccharomyces cerevisiae gene BTN1, encodes a 408 amino acid putative integral membrane protein, which is 39% identical and 59% similar to the human Cln3p, whose mutant forms are responsible for Batten's disease and for a diminished degradation of mitochondrial ATPase synthase subunit c. Disruption experiments established that Btn1p is not essential for viability, mitochondrial function, or degradation of mitochondrial ATP synthase in yeast. PMID- 9219334 TI - Secretion of mouse alpha-amylase from Kluyveromyces lactis. AB - We constructed two mouse alpha-amylase secretion vectors for Kluyveromyces lactis using the well-characterized signal sequence of the pGKL 128 kDa killer precursor protein. Both PHO5 and PGK expression cassettes from Saccharomyces cerevisiae directed the expression of mouse alpha-amylase in YPD medium at a similar level of efficiency. K. lactis transformants secreted glycosylated and non-glycosylated alpha-amylase into the culture medium and both species were enzymatically active. The K. lactis/S. cerevisiae shuttle secretion vector pMI6 was constructed, and K. lactis MD2/1(pMI6) secreted about four-fold more alpha-amylase than S. cerevisiae YNN27 harboring the same plasmid, indicating that K. lactis is an efficient host cell for the secretion and production of recombinant proteins. PMID- 9219335 TI - Deregulation of CLN1 and CLN2 in the Saccharomyces cerevisiae whi2 mutant. AB - Wild-type cells of the budding yeast Saccharbmyces cerevisiae arrest in G1 upon nutrient exhaustion. Cell cycle arrest requires the WHI2 gene since whi2 mutants continue to divide and become abnormally small as nutrients are depleted. Here we show that CLN1 and CLN2 transcript levels in a whi2 strain are higher during exponential growth, and persist longer upon starvation, than in an isogenic wild type strain. In contrast to CLN1 and CLN2, CLN3 levels declined only at very high cell density and were unaffected by the whi2 mutation. Elevated CLN expression is sufficient to explain the whi2 phenotype since ectopic expression of CLN1 in a nutrient-depleted culture caused cells to continue dividing and interfered with the acquisition of heat resistance. These observations show that, either directly or indirectly, Whi2 negatively regulates G1 cyclin expression. Interestingly extremely high levels of Cln1 induced filamentous growth upon nutrient deprivation, suggesting a direct connection between G1 cyclin activity and morphological responses to poor nutrient conditions. PMID- 9219336 TI - Constitutive flocculation in Saccharomyces cerevisiae through overexpression of the GTS1 gene, coding for a 'Glo'-type Zn-finger-containing protein. AB - The product of the cloned GTS1 gene is characterized by structural features found in transcription factors. It contains one Zn-finger motif (CXXCX16CXXC) situated in the N-terminal end with a high degree of homology to the newly identified 'Glo' family of Zn-finger proteins (Ireland et al., 1994, EMBO J. 13, 3812-3821). The C-terminal end of the protein is characterized by poly (Ala-Gln) and poly-Gln stretches. Poly-Gln are part of trans-acting motifs in known transcription factors. Overexpression of the GTS1 gene results in constitutive flocculation. Whole cell electrophoretic mobility and hydrophobicity of GTS1 overexpressing cells was respectively lower and higher relative to control cells. GTS1-induced flocculation is hardly sensitive to mannose in contrast to FLO1-determined flocculation. Overexpression of the GTS1 gene in a flo1 background does not abolish flocculation, suggesting that the FLO1 gene is not linked with the GTS1 gene in a 'flocculation pathway'. PMID- 9219337 TI - Identification and preliminary characterization of p31, a new PSTAIRE-related protein in fission yeast. AB - One of the defining characteristics of the catalytic subunit of the cyclin dependent protein kinases (cdks) is the so-called PSTAIRE motif. Western blots of fission yeast cytosolic extracts using a monoclonal antibody against the PSTAIRE peptide revealed two bands at 34 kDa (p34cdc2) and 31 kDa (p31). Polyclonal antibodies to the C-terminus of p34cdc2 or to the full-length protein recognized the 34 kDa band but not p31. Overexpression of the cdc2+ gene resulted in the increase of the 34 kDa band but not p31. Like p34cdc2, the level of p31 revealed no obvious cell cycle regulation but the protein was present in spores where p34cdc2 was barely detectable. p31 expression was unaffected by removal of either phosphate or ammonium from the growth medium, although the level of p34cdc2 was reduced in the absence of phosphate. p31 was not associated with cyclin B, nor was it adsorbed to p13suc1 Sepharose beads, two characteristics of p34cdc2. p31 did, however, interact with p15, the starfish homologue of p13suc1. p31 was present in cells in which cdc2+ was replaced by its budding yeast homologue CDC28. When fission yeast cytosolic extracts were subjected to gel filtration chromatography, p31 eluted in two peaks, one at approximately 100 kDa, the other at approximately 30 kDa. We conclude that p31 is a novel fission yeast PSTAIRE protein and therefore, potentially, a new cdk. PMID- 9219338 TI - Characterization of the checkpoint gene RAD53/MEC2 in Saccharomyces cerevisiae. AB - Saccharomyces cerevisiae cells carrying mutations in RAD53/MEC2 fail to arrest in the S phase when DNA replication is blocked (the S/M checkpoint) or in the G2 phase when DNA is damaged (the G2/M checkpoint). We isolated and determined the DNA sequence of RAD53 and found that it is identical to the SPK1 gene previously identified by Stern et al. (1991). In addition to its checkpoint functions, we show here that RAD53 is essential for cell viability because null mutants are inviable. Weak genomic suppressors of the essential function do arise frequently, though they do not suppress the checkpoint defects of the null mutant. This genetically separates the essential and checkpoint functions. We show genetically that the protein kinase domain is essential for all RAD53-dependent functions tested because a site-specific mutation that inactivates the protein kinase activity results in a mutant phenotype indistinguishable from that of a null mutant. Overexpression of RAD53, or its kinase domain alone, resulted in a delay in cell-cycle progression that required the intact kinase function. The cell cycle delay did not require any of the checkpoint genes tested (e.g. rad9 or mecl), indicating that the cell-cycle delay is either unrelated to the checkpoint responses, or that it occurs constitutively because RAD53 acts further downstream of the checkpoint genes tested. Finally, elimination of sequences in the promoter region of RAD53 revealed complex regulatory elements. PMID- 9219340 TI - Applications of the long and accurate polymerase chain reaction method in yeast molecular biology: direct sequencing of the amplified DNA and its introduction into yeast. AB - A DNA fragment longer than 10 kb can be amplified by the long and accurate polymerase chain reaction (LA-PCR) method. We demonstrate here applications of this technique in molecular biological studies of Saccharomyces cerevisiae. We have shown that DNA fragments amplified by LA-PCR can be directly used as a template in the chain-termination sequencing protocol, making it possible to quickly identify the DNA insert of yeast genomic library clones. We have also shown that the amplified yeast DNA can easily be introduced into yeast by co transformation with linearized vector DNA. Overlapping DNA between the amplified yeast fragment and the vector must be more than 20 bp long in order to obtain 90% or more correct recombinant plasmids. These results suggest that simple amplification of yeast clones by LA-PCR can replace the previous procedures of yeast clone recovery, consisting of transformation of Escherichia coli, propagation of plasmids in E. coli and preparation of plasmid DNA. PMID- 9219339 TI - Cloning of the multicopy suppressor gene SUR7: evidence for a functional relationship between the yeast actin-binding protein Rvs167 and a putative membranous protein. AB - The rvs161 and rvs167 mutant cells exhibit several identical phenotypes including sensitivity to several different growth conditions and morphological defects such as alteration of the actin cytoskeleton and budding patterns. The selection of genes that, when overexpressed, are able to suppress the reduced viability upon carbon starvation of the rvs167 mutant strain, has allowed the cloning of the SUR7 gene (Accession Number Z46729x11). We showed that the suppressive ability of the overexpressed SUR7 gene concerns all the rvs167 phenotypes. However, this suppression is only partial since the rvs167-suppressed strain is not of wild type phenotype. Moreover, SUR7 is also able to suppress partially the phenotypes exhibited by the rvs161 and rvs167 and rvs161 mutant strains. The SUR7 gene encodes a putative integral membrane protein with four transmembrane domains. Furthermore, sequence comparisons revealed that Sur7p and two other proteins, Yn1194p and Yd1222p, present significant sequence and structural similarities. Taken together, these results strongly suggest that the Rvs161 and Rvs167 proteins act together in relation with Sur7p. Moreover, the putative transmembranous character of Sur7p suggests a membrane localization of the Rvs function, a localization which is consistent with the different rvs phenotypes and the actin-Rvs167p interaction. PMID- 9219341 TI - Sequence analysis of the Candida albicans ADE2 gene and physical separation of the two functionally distinct domains of the phosphoribosylaminoimidazole carboxylase. AB - An ADE2 genomic clone from the pathogenic fungus, Candida albicans, was isolated by complementation of an Escherichia coli purK mutant and the gene was analysed by DNA sequencing. A 1707 bp open reading frame was identified encoding a polypeptide of 569 amino acids with significant homology to all the known yeast ADE2 genes. Sequence homology to both the E. coli purE and purK genes suggests that the C. albicans ADE2 gene is the result of an evolutionary fusion. The amino acid sequence comparison showed that the N-terminal domain of the Ade2 protein has a 52.5% identity to purK, whereas the C-terminal domain has a distinct 64.3% identity to purE. In order to establish the functional relationship of these two regions, deletion mutants of the Ade2 protein were prepared by recombinant expression of the functional domains, which were tested by complementation of their respective E. coli auxotrophs. PMID- 9219342 TI - Cloning and characterization of the KlDIM1 gene from Kluyveromyces lactis encoding the m2(6)A dimethylase of the 18S rRNA. AB - The KlDIM1 gene encoding the m2(6)A rRNA dimethylase was cloned from a Kluyveromyces lactis genomic library using a PCR amplicon from the Saccharomyces cerevisiae ScDIM1 gene as probe. The KlDIM1 gene encodes a 320-amino acid protein which shows 81% identity to ScDim1p from S. cerevisiae and 25% identity to ksgAp from Escherichia coli. Complementation of the kasugamycin-resistant ksgA-mutant of E. coli lacking dimethylase activity demonstrates that KlDim1p is the functional homologue of the bacterial enzyme. Multiple alignment of dimethylases from prokaryotes and yeasts shows that the two yeast enzymes display distinctive structural motives including a putative nuclear localization signal. PMID- 9219343 TI - Labeling proteins at high specific activity using N-succinimidyl 4 [18F](fluoromethyl) benzoate. AB - High effective specific activity N-succinimidyl 4-[18F](fluoromethyl)benzoate was prepared using a reversed phase HPLC procedure. Reversed phase HPLC removed several additional impurities not removed by normal phase HPLC, thereby increasing the effective specific activity. Small amount (< 100 micrograms) of sensitive proteins such as erythropoietin can be labeled with this reagent in high yield without aggregation. PMID- 9219344 TI - The use of associated particle timing based on the D + D reaction for elemental analysis of bulk samples such as the human body. AB - The use of associated particle timing based on the D + D reaction has been demonstrated for elemental analysis of bulk samples such as the human body. The neutron energy of 2.8 MeV eliminates the background from organic matrices. The nanosecond timing of a HPGe detector renders it possible to identify the spatial origin of the measured gamma radiation limiting the sensitive area to a single pixel. By this technique the background could be reduced by a factor of > or = 1000, but the present set-up has achieved an effective factor only in the range 20-100, due to losses in the generation of timing signals. The very clean gamma spectra obtained permit the use of high efficiency scintillation detectors. Sensitivities for measuring Al, Ti, and Fe are presented at an extrapolated dose of 10 mSv. PMID- 9219345 TI - Oxidative stress effect on the integrity of lipid bilayers is modulated by cholesterol level of bilayers. AB - Large unilamellar vesicles (120-160 nm) composed of egg phosphatidylcholine (egg PC) containing approximately 22 wt% of polyunsaturated fatty acids (PUFA) and various mol% (0, 10, 22, or 45) of cholesterol were exposed to oxidative stress. The hydrophilic azo compound 2,2'-azobis-(2-amidinopropane)2HCl (AAPH) which was thermally decomposed to produce a constant flux of peroxy radicals was the source of the oxidative stress (< or = 48 h incubation at 37 degrees C). Cholesterol loss following the oxidation was up to 33%, while PUFA were more extensively damaged; loss was up to 52, 88, and 100% for C-18:2, C-20:4, and C-22:6, respectively. (ii) Oxidizability of cholesterol when quantified in absolute amount was three-fold higher when its level was 45 mol%. The interrelationship between bilayer structure, especially its lateral organization and free volume, and lipid peroxidation are discussed. Differential scanning calorimetry of oxidized multilamellar vesicles lacking cholesterol revealed that a high level of oxidative damage to egg phosphatidylcholine PUFA resulted in the loss of the gel to liquid-crystalline phase transition of egg PC (broad peak at around -8 degrees C). PMID- 9219346 TI - Novel cationic liposomes for DNA-transfection with high efficiency and low toxicity. AB - Liposomes containing the natural cationic amphiphile, sphingosine and some of its derivatives were used for transfection of DNA in vitro. Multilamellar liposomes comprised of dioleoylphosphatidylethanolamine (DOPE), different sphingosine derivatives, and diacylglycerols with varying fatty acid chains, preincubated with DNA, transfected efficiently the KK-1 murine granulosa cells. Most efficient transfection on this cell line was achieved with liposomes composed of phytosphingosine, DOPE, and dioctanoylglycerol (DC8G) (64:31:4.8, molar stoichiometry), which gave expression of the transfected gene 2-10-fold higher than the commercial reagent Lipofectin. At higher doses the new liposomes also caused markedly less cell death of KK-1 cells. On COS-7 cells these liposomes showed slightly, but significantly lower transfection, of approximately 70%, of that gained with Lipofectin. The murine Sertoli cells, MSC-1, selectively resisted transfection by the sphingosine derivative based liposomes tested, giving only 11-14% of the expression detected in Lipofectin transfected cells of the same line. In conclusion, the novel liposomes formulated offer an effective, technically easy and economical method of transfection for a variety of cultured cell lines. PMID- 9219347 TI - New nucleoside-5'-alkylphosphonophosphates and related compounds containing 2' deoxycytidine, thymidine and adenosine as nucleoside component. Syntheses and their effects on tumor cell growth in vitro. AB - Recent studies have shown that phosphono analogs of cytidine-5'-diphosphate diacylglycerol (CDP-DAG) possessing a structurally modified lipid moiety exhibit antiproliferative activity in vitro. As an extension of our previous work we tried to elucidate whether the presence of the cytidine component is necessary for cytostatic activity. In this context we have synthesized similarly structured nucleoside-phospholipid conjugates containing nucleoside components other than cytidine, which also do not exhibit cytostatic properties as such. The compounds include 5'-alkyldiphosphates and 5'-alkylphosphonophosphates of 2'-deoxycytidine, thymidine and adenosine with different alkyl chain length as well as selected 3 hexadecyl-2-chloro-2-deoxyglycero-(1)-diphosphates and -phosphonophosphates of these nucleosides. The chemical structures of the newly synthesized nucleoside phospholipid conjugates were confirmed by fast atom bombardment (FAB) and electrospray ionization (ESI) mass spectrometry. It was found that these compounds also inhibit the cell growth of different human cell lines, i.e. the presence of the cytidine component is not a necessary prerequisite for the antiproliferative activity of these nucleoside-phospholipid conjugates. PMID- 9219348 TI - Increased levels of lipid oxidation products in low density lipoproteins of patients suffering from rheumatoid arthritis. AB - 9-Hydroxy-10,12-octadecadienoic acid (9-HODE) and 13-hydroxy-9,11-octadecadienoic acid (13-HODE) are accumulated in the low density lipoproteins of patients suffering from rheumatoid arthritis for a factor of 20-50 compared to healthy individuals of the same age. Both acids, derived by lipid peroxidation of linoleic acid, induce the release of interleukin 1 beta. The latter induces bone degression. The genesis of 9- and 13-HODE seems therefore to be an important factor in the development and progression of rheuma; in addition 9-HODE was reported to be a stimulus of inflammation, comparable to leukotrienes. PMID- 9219349 TI - The concentrations of vitellogenin (vitellin) and protein in hemolymph, ovary and hepatopancreas in different ovarian stages of the freshwater prawn, Macrobrachium rosenbergii. AB - The objectives were to measure the concentrations of vitellogenin (vitellin) and protein in hemolymph, ovary, and hepatopancreas of the freshwater prawn, Macrobrachium rosenbergii, in different stages of ovarian development. The ovarian development of M. rosenbergii was classified into five developmental stages (Stages I-V). Vitellogenin concentrations increased in the hemolymph of prawns in the early stages of ovarian development (Stage I or II) and were maintained at high levels until Stage V. There was no close association between ovarian development and the concentrations of circulating vitellogenin. Concentrations of protein in hemolymph and hepatopancreas remained constant during various stages of ovarian development. The ovarian stages closely correlated with the gonadosomatic index, the concentrations of ovarian vitellin and protein, respectively. Vitellogenin levels in hepatopancreas remained very low in different stages of the prawn although the highest levels were observed in Stage IV. No close association between hemolymph vitellogenin and ovarian vitellin was observed. The increase of vitellogenin concentration in hemolymph occurred earlier than vitellin content in ovary. PMID- 9219350 TI - Temperature and neural development of the Atlantic herring (Clupea harengus L.). AB - Embryos of Atlantic herring (Clupea harengus L.) from the Buchan (Northern North Sea) stock were incubated from fertilisation until hatching at temperatures of 5, 8, 12, and 15 degrees C. The relative timing of development of the Kolmer-Agduhr (KA) neurons, the posterior lateral line nerve, the motor neurons, and myotubes were determined with respect to somite stage of the embryo. Development of the KA neurons, the lateral line nerve, and the myotubes was similar at all temperatures. In contrast, timing of outgrowth of the motor neuron axons with respect to somite stage was earlier at higher (> or = 12 degrees C) than at lower temperatures (< or = 8 degrees C) although it reached a similar point at all temperatures by the 58-somite stage. Our hypothesis to explain these observations is that delayed motor axon outgrowth in the lower temperature groups is probably due to a delay in a signalling interaction between motor neurons and the somite. PMID- 9219351 TI - Variation in response to growth factor stimuli in satellite cell populations. AB - Variation in response to growth factor stimuli in myogenic satellite cell populations was investigated using a clonal-derived satellite cell culture system. Satellite cell clones were established from one muscle from one individual animal. One clone ("Early") which reached confluence on day 19 and one clone ("Late"), which reached confluence on day 29, were chosen for further examination. In previous studies, these two clones were found to differ in their growth rates in serum-containing medium. In the present study, the influence of growth factors on the proliferation of the two clones was compared in serum-free defined medium. Although basic fibroblast growth factor (bFGF), insulin-like growth factor-I (IGF-I), insulin, and platelet-derived growth factor-BB (PDGF-BB) stimulated proliferation of both clones, the Early clone was more responsive to all growth factors tested than the Late clone (P < or = 0.05). The Early clone was also more responsive to the proliferative and differentiative depressing effects of administered transforming growth factor-beta (P < or = 0.05). Examination of properties of the PDGF, FGF, and IGF-I receptors on these two clones revealed no differences in either dissociation constants or receptor numbers (P > or = 0.05). The results suggest that there is heterogeneity in satellite cell response to growth factors. PMID- 9219352 TI - Identification of neuronal isozyme specific residues by comparison of goldfish aldolase C to other aldolases. AB - A 2061 bp cDNA encoding a goldfish (Carassius auratus) aldolase was isolated from a goldfish brain library. The deduced 362 amino acid sequence is more similar to vertebrate brain (aldolase C) and muscle aldolases (aldolase A) than to the liver isozymes (aldolase B). Northern blot analysis indicates strong expression of the mRNA in brain but not in liver or muscle, which indicates that this is aldolase C rather than aldolase A. Analysis of all known vertebrate aldolase amino acid sequences reveals five residues; Leu-57, Arg-314, Thr-324, Glu-332, and Gly-350 that are present exclusively in aldolase Cs. The goldfish clone possesses all five residues. The residues are primarily located in the carboxyl-terminal region of the enzyme and may play a role in determining the neuronal isozyme-specific properties of the enzyme. Furthermore, the existence of an aldolase C in a teleost fish has implications with respect to the timing of genome duplication events that are thought to have been critical in vertebrate evolution. PMID- 9219353 TI - Drinking in Atlantic salmon presmolts and smolts in response to growth hormone and salinity. AB - Drinking rate in freshwater Atlantic salmon presmolts (about 0.1 ml/kg/h) was unaffected by daily injections of ovine GH (50 micrograms/fish) for a week but upon transfer to sea water an immediate and full drinking response was developed compared to saline treated fish (3.34 +/- 0.16 vs. 2.23 +/- 0.27 ml/kg/h). Smolting did not affect drinking rates in freshwater but after 7 days in sea water, salmon smolts imbibed 3.88 +/- 0.25 ml/ kg/h, significantly higher than the rate for saline injected presmolts (p < 0.05, one-way ANOVA), but not significantly different from oGH treated presmolts. Smolting and oGH treatment were without effect on plasma Na+ levels in freshwater fish and 7 days after transfer to sea water both groups showed a better regulation of plasma Na+ levels compared to saline treated presmolts. Atlantic salmon smolts showed higher levels of plasma Cl- than presmolts in freshwater, and after 7 days in sea water, both oGH presmolts and smolts showed significantly lower levels of plasma Cl- than saline injected presmolts. GH treatment in freshwater presmolts improved hypoosmoregulatory capacity following transfer to seawater and these results are discussed in relation to the physiology of smolting, and control of drinking. PMID- 9219354 TI - Cold- and ouabain-resistance of renal Na,K-ATPase in cold-exposed and hibernating jerboas (Jaculus orientalis). AB - The temperature dependence and the ouabain sensitivity of Na,K-ATPase was examined in the nephron of normal, cold-exposed, and hibernating jerboas. The transport and hydrolytic activity of renal Na,K-ATPase displayed similar temperature dependence in rats and normal jerboas. Cold-resistance of Na,K-ATPase appeared in cold-exposed jerboas and further increased during hibernation. Three subpopulations of Na,K-ATPase displaying very high (Ki approximately 10(-13) M), high (Ki approximately 10(-9) M) and low sensitivity to ouabain (Ki approximately 10(-6) M) were detected in the thick ascending limb and collecting duct of jerboas. In thick ascending limbs, the subpopulation of very high sensitivity to ouabain disappeared in cold-exposed animals, which accounted for the previously reported decrease in Na,K-ATPase activity. In collecting ducts of cold-exposed animals, the subpopulation of very high sensitivity to ouabain also disappeared, but the resulting decrease in activity was overbalanced by the appearance of the subpopulation of high sensitivity. PMID- 9219355 TI - Specific dynamic action of a large carnivorous lizard, Varanus albigularis. AB - Varanus albigularis inhabits grasslands of southern and eastern Africa and experiences months of fasting during the dry season (May-December) followed by voracious feeding during the wet season (January-April). Previous studies have found that sit-and-wait foraging snakes, which also experience long intervals between large meals, exhibit unprecedented increases in post-feeding metabolism, which reflects the added cost of up-regulating a previously quiescent gut and digesting a large meal. Hence we measured pre- and post-prandial oxygen consumption rates (VO2) of adult V. albigularis in order to observe whether they exhibit similarly large metabolic responses to digestion as sit-and-wait foraging snakes. Following the consumption of meals consisting of ground turkey and snails, hard-boiled eggs, or juvenile rats, lizards rapidly increased their VO2 to peak within 24-27 hr at 7-10 times pre-feeding values (mean = 0.035 mL O2.g 1.h-1). During the 60-90 hr of significantly elevated VO2, the extra oxygen consumed (the specific dynamic action) represented an energy expenditure of 830 1260 kJ. For meals that were fully digested, specific dynamic action equalled 24% of ingested energy. The magnitudes of V. albigularis post-prandial metabolic responses are similar to those previously observed for sit-and-wait foraging snakes. Like sit-and-wait foraging snakes, V. albigularis may also down-regulate intestinal performance during their months of fasting (suggested by their relatively low standard metabolic rate) and then up-regulate their gut (bearing its high energetic cost) upon feeding. PMID- 9219356 TI - Binding of 125I-insulin on capillary endothelial and myofiber cell membranes in normal and streptozotocin-induced diabetic perfused rat hearts. AB - A heart-perfusion technique was employed to measure 125I-insulin binding on capillary endothelial and myocyte cell membranes in Sprague-Dawley rats. Animals were anesthetized, and the anterior chest wall excised to expose the mediastinal contents. The right and left superior and inferior venae cavae were dissected and tied, and another tie was passed around the aorta. A polyethylene catheter was introduced into the aortic lumen from cephalad to caudad to sit with its tip above the aortic valve. Another catheter was introduced into the cavity of the right atrium and both were anchored by sutures. Oxygenated Ringer-Lock buffer containing 20 mM/L K+ and 125I-insulin was perfused at a rate of 1 mL/min via the aortic catheter. Concomitantly, the distal ascending aorta and venae cavae were ligated. The effluent was collected from the right atrial catheter at the same infusion rate. Animals were divided into two groups, the normal group and streptozotocin-induced diabetic group. Heart perfusion was done on both groups either without or after treatment with detergent (CHAPS) to remove the capillary endothelial lining. A physical model for 125I-insulin sequestration as a ligand to its receptors on endothelial and/or myocyte plasma membranes was proposed. The model described a reversible binding of ligand on cellular surface receptor concentration to fit a conservation equation and a first order Bessel function. The binding constants (kn), reversal constants (k-n), dissociation constants kd = k-n/kn, and residency time constants tau = 1/k-n of 125I-insulin in normal untreated, normal CHAPS-treated, diabetic untreated, and diabetic CHAPS-treated hearts were estimated using a theoretically generated curve-fit to the data. Since insulin receptor binding on the capillary endothelial cell surfaces may serve to transport insulin from the intravascular to the subendothelial space, and since streptozotocin-induced diabetes was shown to diminish receptor autophosphorylation and kinase activity and hence internalization of insulin, then one can conclude the following from the data. In the normal heart, removal of the capillary endothelial lining with CHAPS did not alter kn, k-n, kd, and tau of insulin binding as compared to the normal untreated, whereas in the diabetic untreated heart these constants were altered, compared to the diabetic treated. Furthermore, the kn and k-n values in the diabetic CHAPS-treated hearts were the same as for the normals untreated and CHAPS-treated, respectively. In conclusion, the dissociation constants and residency time constants of all groups indicated the possible existence of two types of insulin receptors: the capillary endothelial cell surface insulin receptors with lower residency time (low affinity receptor or combination of insulin and IGF-1 receptors) and the myocyte plasma membrane insulin receptors with higher residency times (high affinity). PMID- 9219357 TI - Potential for non-shivering thermogenesis in perfused chicken (Gallus domesticus) muscle. AB - The humoral modulation of resting muscle heat production of chickens (Gallus domesticus) was investigated in vitro. The resting distal lower limb was perfused via the popliteal artery at 25 degrees C without erythrocytes at constant flow. The preparation was stable for at least 3 hr, showing a constant oxygen uptake (MO2) and perfusion pressure as well as adequately maintaining muscle energy charge and creatine phosphate: creatine ratio. Noradrenaline (NOR), adrenaline (ADR) and serotonin (5-HT) each caused a dose-dependent rise in perfusion pressure. NOR and ADR evoked increased MO2 at low doses eventually followed by decreased MO2 at higher agonist concentrations. 5-HT gave smaller but qualitatively similar MO2 effects. The actions of 50 nM NOR were blocked by prazosin (10 microM) and nitroprusside (0.5 mM), but not altered by propranolol (10 microM). NOR-induced stimulatory MO2 changes in the presence of pharmacological concentrations (1 microM) of glucagon were more pronounced and the thermogenic concentration range of NOR was increased. Taken together, these in vitro findings demonstrate a potential for vasoconstrictor-controlled muscle nonshivering thermogenesis in birds as in marsupials and mammals, suggesting that vascular control of muscle MO2 may be a widespread biological mechanism. The possible implications of these findings for avian nonshivering thermogenesis are discussed. PMID- 9219358 TI - Immunohistochemical demonstration of bleeding in decomposed bodies by using anti glycophorin A monoclonal antibody. AB - The usefulness of glycophorin A (GPA) as a marker of bleeding was investigated in decomposed bodies by using anti-human GPA monoclonal antibody immunohistochemically. Ninety-one specimens consisting of 37 skin and 54 muscle specimens were obtained from 21 autopsy cases with various degree of decomposition, which ranged from 12 h up to 2-3 months after death. The presence or absence of the bleeding in the specimens was evaluated macroscopically and was divided as follows: (1) specimens without bleeding (31 specimens), (2) specimens with bleeding (15 specimens), and (3) suspect specimens (45 specimens), in which the bleeding was not clear. By a peroxidase-labeled streptavidin-biotin method, positive reaction products for GPA were observed only within the blood vessels in the specimens without bleeding. On the other hand, in the specimens with bleeding, positive reaction products for GPA were seen not only within the blood vessels but also the extravascular tissues. Therefore, a specimen can be diagnosed as bleeding when GPA is distributed both within blood vessels and tissue outside the vessels. In application of GPA to 45 suspect specimens, 42 specimens (93%) were distinguished from the specimens with bleeding or without bleeding. These results prove that GPA is very useful as a marker of bleeding. The detection of GPA by the immunohistochemical method will help to differentiate between bleeding and hemoglobin (Hb) diffusion from blood vessels in a decomposed body. PMID- 9219359 TI - Fractures of the base of the skull in charred bodies--post-mortem heat injuries or signs of mechanical traumatisation? AB - Based on a recent case, in which an expert opinion had to be prepared, the question was investigated if fractures of the base of the skull can result from the influence of heat on the human skull. Neither the retrospective analysis of autopsy records nor the prospective examination of charred bodies revealed any cases with heat-induced fractures of the base of the skull. Observation of cremations showed that the changes caused by the fire followed certain rules: fractures of the calvaria were seen after approximately 20 min; the base of the skull became exposed after about 45 to 60 min. In none of the 20 cremations watched could any fractures of the base of the skull be detected. PMID- 9219360 TI - Infrared fluorescent detection of D1S80 alleles. AB - A genetic locus D1S80 containing a variable number of tandem repeats (VNTR) has been used extensively in forensic analysis and paternity testing. In the current research, the D1S80 locus was amplified using polymerase chain reaction (PCR) technology and the alleles detected using a high sensitivity infrared (IR) fluorescence automated DNA sequencer. IR-labeled amplification products were generated using oligonucleotide primers which were covalently linked to an infrared fluorescent dye (IRD41) at the 5'-end. Human genomic DNA (1.0 ng or less) isolated from blood and various simulated forensic samples was successfully amplified using this technology. Allelic bands were detected by incorporation of the IR fluorescent dye into PCR products. Both Long Ranger and polyacrylamide denaturing gels permitted clear resolution of individual alleles that differ by only one repeat unit. In the smaller gels a separation distance of only 15 cm allowed separation of the alleles in less than 2 h from sample loading to visualization. This system combines IR fluorescence chemistry and laser technology thus eliminating the need for post-electrophoretic gel handling for the detection of D1S80 alleles. Real-time detection is valuable for immediate visualization of the data and the alleles are displayed as familiar autoradiogram like images which can also be analyzed by computer. By loading a 64-lane gel twice it is possible to type at least 120 samples in 1 day using a single gel. PMID- 9219361 TI - Epidemiology of suicide in elderly people in Madrid, Spain (1990-1994). AB - The progressive aging of the population as a whole, the frequent appearance of degenerative diseases, and the greater frequency of suicide among persons older than 65 years than in younger age groups, are worrisome issues that deserve investigation. The aim of this study was therefore to analyze different epidemiological and social factors that influence suicide behaviour in elderly subjects in Madrid (Spain) during a 5-year period from January 1990 to December 1994. Post-mortem reports on all deaths that were examined at the Institute of Forensic Medicine in Madrid were studied. All cases of suicide in subjects aged > or = 65 years during these years (N = 461) were studied through the autopsy records and information from the coroner's inquest. Variables corresponding to demographic, clinical and interpersonal factors, method of suicide, scene of death, season, month and time of suicide were registered. In both sexes, jumping from a height was the most frequent method (63.6%). Family members had noted symptoms of depression in almost half of the cases (49.5%). Coexisting physical disorders were present in 68.9% of the subjects. Health care professionals have an important role to play in-suicide prevention. PMID- 9219362 TI - Molecular characterization of a defensin gene from the mosquito, Aedes aegypti. AB - Insect immune proteins, defensins, are inducible anti-Gram-positive bacterial peptides. We report here the identification of two defensin genes from the mosquito, Aedes aegypti, which encode a large 541 bp transcript (AaDef Ala) and a small 473 bp transcript (AaDef Asm). The cDNA corresponding to AaDef Ala was cloned, sequenced, and compared with the previously reported AaDef Asm cDNA. The AaDef Ala gene was isolated through genomic library screening and characterized. It putative regulatory region contains a 64 bp intron, a TATA box and a putative arthropod initiator. Two 150 bp long direct and several palindromic repeats are present in this sequence. Similar to other insect immune peptide genes, the AaDef Ala gene contains numerous putative regulatory motifs with impressive similarity to elements of vertebrate acute phase response protein genes. PMID- 9219363 TI - Transposition of the Hermes element in embryos of the vector mosquito, Aedes aegypti. AB - Using a plasmid-based transpositional recombination assay in vivo, we have demonstrated that Hermes, a short inverted repeat type transposable element from Musca domestica, can transpose in Aedes aegypti embryos. Hermes transpositions in Ae. aegypti have all the characteristics observed during Hermes transposition in its host M. domestica and in related species. These characteristics include an absolute dependence on the expression of the Hermes transposase and a preference for the integration site GTNCAGAC (P < 0.05). In addition, the rate of Hermes transposition in Ae. aegypti (0.286 transpositions per 10,000 donor plasmids screened) was comparable to that observed in Drosophila melanogaster under similar conditions. These results suggest that Hermes can be developed into a gene vector and genetic engineering tool for Ae. aegypti and related mosquitoes. PMID- 9219364 TI - Isolation and characterization of CYP6B4, a furanocoumarin-inducible cytochrome P450 from a polyphagous caterpillar (Lepidoptera:papilionidae). AB - Papilio glaucus (tiger swallowtail) is a generalist that rarely encounters plants containing furanocoumarins yet is constitutively capable of metabolizing low levels of these highly toxic allelochemicals. In larvae of this species, metabolism of linear (xanthotoxin, bergapten), and angular (angelicin, sphondin), furanocoumarins can be induced up to 30-fold by the presence of xanthotoxin in their diet. Degenerate primers corresponding to conserved amino acid sequences in three insect P450s, Musca domestica (CYP6A1), Drosophila melanogaster (CYP6A2) and Papilio polyxenes (CYP6B1), were used to clone xanthotoxin-induced P450 transcripts from P. glaucus larvae by a reverse transcription-polymerase chain reaction (RT-PCR) strategy. Positive clones encoding the highly conserved F--G-R C-G P450 signature motif were used to isolate a full-length CYP6B4v1 cDNA from a P. glaucus xanthotoxin-induced cDNA library. Sequence comparisons indicate the P. glaucus CYP6B4v1 protein sequence is 63% and 61% identical, respectively, to the P. polyxenes furanocoumarin-inducible CYP6B1v1 and CYP6B3v1 proteins. Northern analysis indicates that CYP6B4 and related transcripts are highly induced in response to xanthotoxin. Baculovirus-mediated expression of the CYP6B4v1 protein in lepidopteran cell lines demonstrates that this P450 isozyme metabolizes isopimpinellin, imperatorin, and bergapten at high rates, xanthotoxin and psoralen at intermediate rates and angelicin, sphondin, and trioxsalen only at very low rates. PMID- 9219365 TI - A specific prostaglandin E2 receptor and its role in modulating salivary secretion in the female tick, Amblyomma americanum (L.). AB - Prostaglandins of the 2-series (e.g. PGE2) are typically synthesized from arachidonic acid (AA) after AA is released from cellular phospholipids after activation of an intracellular phospholipase A2 (PLA2). Treatment of isolated salivary glands with PLA2 inhibitor oleyloxyethyl phosphorylcholine (OPC) or prostaglandin synthetase inhibitors reduced dopamine-induced fluid secretion and cyclic AMP (cAMP) levels in isolated salivary glands. PGE2 and its analog, 17 phenyl trinor PGE2, partly reversed the inhibition of secretion and cAMP level by OPC, suggesting that prostaglandins may have an autocrine effect in modulating tick salivary gland function. A specific PGE2 receptor was identified in the plasma membrane fraction of the salivary glands. The receptor exhibits a single, high affinity PGE2 binding site with a KD approximately 29 nM, is saturable, reversible, and specific for PGE2 and coupled to a cholera toxin-sensitive guanine nucleotide regulatory protein. Assay of adenylate cyclase activity in salivary gland membranes showed that PGE2 neither stimulated nor inhibited adenylate cyclase activity, indicating that the PGE2 effects on cAMP levels and possibly secretion are indirect, and that the PGE2 receptor stimulates an alternate "second messenger" pathway. PMID- 9219366 TI - Expression and characterization of a lepidopteran general odorant binding protein. AB - Olfaction in months involves the transport of volatile, hydrophobic odorant molecules through the aqueous interior of the antennal sensory hairs by soluble odorant binding proteins. Two subfamilies of the 17 kDa general odorant binding proteins, GOBP1 and GOBP2, are 47-57% identical to each other and 21-57% to the pheromone binding proteins (PBPs); identity within a GOBP subfamily exceeds 78% in all lepidopteran species examined. However, the ligands for GOBPs are unknown. In order to investigate odorant specificities of GOBPs, recombinant proteins were expressed in Escherichia coli using PCR-prepared expression cassettes based on the cDNA sequences of GOBP1 and GOBP2 from Manduca sexta. Both soluble and insoluble recombinant GOBPs (rGOBPs) were obtained, and the inclusion body GOBPs were solubilized, refolded and purified. The soluble and refolded rGOBPs were purified by preparative isoelectric focusing (IEF), gel filtration, and finally by ion-exchange fast protein liquid chromatography (FPLC). Only rGOBP2, but not rGOBP1, was crossreactive with an anti-GOBP2 (Antheraea polyphemus) antiserum. rGOBP2, but not rGOBP1, could be photoaffinity labelled by the diazoacetate pheromone analog [3H]-6E, 11Z, 16:Dza. For rGOBP2, plant odors such as (Z)-3 hexen-1-ol (3Z-6:OH), geraniol, geranyl acetate, and limonene showed significant competition for binding; binding specificity was sensitive to pH and to salt concentrations. Circular dichroism (CD) confirms that, as with the pheromone binding proteins, GOBP2 is predominantly alpha-helical. Although the characterization of rGOBP1 has resisted analysis, rGOBP2 is readily prepared and studied. We suggest that GOBP2 may be broadly tuned to a class of "green" and floral odors. PMID- 9219367 TI - Novel antibacterial peptides isolated from a European bumblebee, Bombus pascuorum (Hymenoptera, Apoidea). AB - We present here the isolation and characterization of four antimicrobial peptides produced by a European bumblebee Bombus pascuorum. A 51-residue insect defensin was characterized which, like the Apis mellifera defensins, had a highly conserved 12-residue extension to its C-terminal compared to defensins from other insects. Monoisotopic mass analysis of the C-terminal of B. pascuorum defensin confirmed that this molecule was C-terminally amidated. This defensin showed strong anti-Gram-positive activity and some anti-fungal activity; also, in contrast to other insect defensins, it showed anti-Gram-negative activity. A 17 residue apidaecin was characterized, showing anti-Gram-negative activity, and differing by a single amino acid substitution from the A. mellifera apidaecin. A 39-residue abaecin was isolated, the largest proline-rich antimicrobial peptide characterized to date, which showed activity against both Gram-negative and Gram positive bacteria. Finally, we isolated an N-terminally blocked molecule, with a molecular mass of 10,122 Da, which showed activity against Gram-negative bacteria only. These characteristics are reminiscent of hymenoptaecin from the honeybee A. mellifera, but a definitive characterization of this molecule awaits further work. No evidence of lysozyme activity was found in the haemolymph of challenged or naive B. pascuorum. PMID- 9219368 TI - Detection of expressed chloramphenicol acetyltransferase in the saliva of Culex pipiens mosquitoes. AB - Mosquito salivary glands play an important role in the transmission of arthropod borne pathogens. The ability to express genes in mosquitoes would be a powerful approach to characterize salivary gland genes, and to reveal important vector determinants of pathogen transmission. Here we report the use of a double subgenomic Sindbis (dsSIN) virus, designated TE/3'2J/CAT, and a packaged Sindbis replicon virus, designated rep5/CAT/26S, to express chloramphenicol acetyltransferase (CAT) protein in the salivary glands and saliva of transduced female Culex pipiens pipiens. Indirect immunofluorescence analysis revealed that salivary glands of these mosquitoes infected with either TE/3'2J/CAT or rep5/CAT/26S virus (4 or 6 days post-infection (p.i.)) were positive for both SIN E1 antigen and CAT protein. Saliva collected from mosquitoes transduced with TE/3'2J/CAT virus contained a unique 25 kDa protein that corresponded to the size of CAT protein. Additionally, CAT activity assays revealed that saliva collected from mosquitoes transduced with either TE/3'2J/CAT or rep5/CAT/26S virus could contain greater than 5.0 x 10(-5) units of CAT enzyme (3.0 x 10(6) CAT trimers). PMID- 9219369 TI - Expression of angiotensin-converting enzyme-related carboxydipeptidases in the larvae of four species of fly. AB - HieACE, a soluble 70 kDa protein related to the angiotensin-converting enzyme (ACE) has recently been identified, characterized and cloned from the adult buffalo fly (Haematobia irritans exigua). HieACE is enzymatically similar to the mammalian ACEs and its predicted amino acid sequence has 42% identity with the mammalian testicular ACEs. In adult H.i. exigua, HieACE expression is restricted to the compound ganglion and posterior midgut, and the maturing male reproductive system. Western blot analysis was used to investigate the expression of HieACE and its homologues in the larvae of H.i. exigua, Drosophila melanogaster, the sheep blowfly (Lucilia cuprina), the Old World screwworm fly (Chrysomya bezziana) and a secondary strike fly, Chrysomya rufifacies. Dipteran ACE homologues of 65 70 kDa were detected in all the larval instars investigated. Most of the immunoreactive proteins were concentrated in the soluble fraction. The first and second larval instars of L. cuprina and C. bezziana appeared to express two ACE homologues. These larvae were also found to secrete (or excrete) the ACE homologue in larval cultures. The presence of ACE-like enzymes in these larvae was confirmed by the measurement of carboxydipeptidase activity that was inhibited by the specific ACE inhibitor, captopril. The tissue distributions of the ACE homologues in the third instar larvae of H.i. exigua and L. cuprina were examined. As in adult H.i. exigua, HieACE was detected in the larval ganglion, but in contrast to the restricted distribution in the adult stage midgut, HieACE was found throughout the digestive system, and in the salivary glands of H.i. exigua larvae. The expression pattern in the gut of L. cuprina larvae was similar despite the differences in diet and habitat. The most striking difference from the adult stage H.i. exigua was the expression of HieACE and its L. cuprina homologues in the hindgut and Malpighian tubules of these larvae. These results suggest that the role(s) played by the dipteran ACE-like enzymes differ between the adult and larval stages. PMID- 9219370 TI - Structure and organization of the Bombyx mori sericin 1 gene and of the sericins 1 deduced from the sequence of the Ser 1B cDNA. AB - The sericin 1 primary transcript of the silkworm Bombyx mori is differentially spliced via a tissue- and developmentally-regulated process. From a middle silk gland cDNA library, we have elucidated the sequence of one of the four mRNAs, the 4.0 kb Ser1B mRNA. Determination of alternative or constitutive exons and intron exon boundaries allowed us to establish the nine exon-eight intron structure of the Ser1 gene. From these and previous data, it was possible to deduce the sequence of the sericins 1 and to predict the secondary structure and physiochemical properties of the different regions of the proteins. PMID- 9219371 TI - Educating workers on the need for a firm, safe footing. PMID- 9219372 TI - Thirst quenchers cool the summer worker. PMID- 9219373 TI - Spectrophotometric studies of complexation of C60 with aromatic hydrocarbons. AB - The equilibrium constants for complexation of C60 with naphthalene, phenanthrene and pyrene in toluene have been determined by UV visible spectroscopy. The magnitude of the equilibrium constants was found to increase with decreasing ionization potential of the donor. Values for complexation enthalpy have been determined for the first time for C60/aromatic hydrocarbons. Well-defined charge transfer (C-T) bands have been observed for complexes of C60 with a variety of aromatic hydrocarbons, with C-T band maxima moving to higher frequency with increasing donor ionization potential. PMID- 9219374 TI - Fourier-transform Raman spectroscopic study of human hair. AB - Fourier-transform Raman microscopic spectra of normal, untreated and bleached hair fibres are presented. Vibrational assignments are made and differences are ascribed to the production of cysteic acid from cysteine. Changes in conformation associated with the disulphide bond in the keratotic component are noted from the v(CSSC) vibrational modes at wave numbers near 500 cm-1. Raman spectra of hair root ends have also been investigated with a diminution in cysteine content being observed. Application of the technique to the biomedical investigation of healthy and diseased hair is proposed. PMID- 9219375 TI - Membership of tobacco industry scientists in scientific societies. PMID- 9219376 TI - Alcohol misuse prevention for young people: a systematic review reveals methodological concerns and lack of reliable evidence of effectiveness. AB - In a systematic review we assessed the methodological quality of evaluations of alcohol misuse prevention programmes for young people, and recorded evidence of effectiveness. Studies were identified through systematic searches of electronic databases; hand searches of all obtained papers, existing reviews and several journals; and mailshots to key organizations, conferences and individuals. Relevant papers were checked and cross-checked by members of the review team, and only those studies with an experimental or quasi-experimental design and positive attributes on a number of other quality criteria were included in the review. After pre-screening over 500 papers which reported prevention programmes, information was systematically abstracted from 155 papers. Only 33 studies merited inclusion in the review, and most of these had some methodological shortcomings. Twenty-one studies reported some significant short- and medium-term reductions in drinking behaviour. Of two studies which carried out longer-term evaluations, only one reported a significant longer-term effect, with small effect sizes. No factors clearly distinguished partially effective from ineffective or harmful prevention programmes. In conclusion, the lack of reliable evidence means that no one type of prevention programme can be recommended. In particular there is a need to carry out well-designed scientific evaluations of the effectiveness of current or new prevention efforts which target young people's alcohol misuse. PMID- 9219377 TI - Alcohol and aviation. AB - Aviation accidents due to alcohol consumption by aircrew appear to be rare, especially in commercial aviation. However, a small proportion of general aviation accidents are attributable to alcohol use by aircrew, and aircrew are not well informed about the metabolism of alcohol and its effects on performance. Furthermore, there is evidence that aircrew performance may be impaired by alcohol consumption even after their Blood Alcohol Concentration has returned to "zero" (i.e. < 5 mg/dl). Accidents caused by impairment of aircrew performance by alcohol may therefore not be attributed to alcohol use at all. Aviation safety relies upon faultless human performance and is thus highly sensitive to alcohol related impairment of performance. This paper provides a review of research regarding aircrew alcohol consumption, impairment of aircrew performance by alcohol, incidence of aviation accidents attributable to alcohol use by aircrew, and other related subjects. PMID- 9219378 TI - The impact of law enforcement activity on a heroin market. AB - It may be argued that seizing large quantities of heroin being imported into the country should decrease its supply and hence increase its price, resulting in a reduction in the quantity of the drug being purchased or consumed. To date, however, there has been no empirical evidence that heroin seizures in Australia have any effect on the price of heroin at street level. This article describes a 2-year research study during which the price and purity of street-level heroin were regularly monitored. It was found that heroin seizures had no effect on the price, purity or perceived availability of heroin at street level. It was further found that admissions to methadone treatment were not affected by the price or perceived availability of heroin or by local arrests for heroin use/possession, nor was any relationship found between these arrests and the price of street level heroin. Nevertheless, two-thirds of those who sought entry to local methadone programmes indicated the price as a reason for stopping using heroin. This paper argues that supply-side law enforcement should only be used as a strategy for maintaining high heroin prices if the demand for heroin can be shown to be price-elastic and, further, that the costs of such a strategy need to be weighted against the benefits. PMID- 9219379 TI - Correlates of college student marijuana use: results of a US National Survey. AB - This study examines which personal student background and college characteristics are associated with marijuana use. A self-administered survey was mailed to a national representative sample of 17592 students at 140 American colleges. One of four (24.8%) students reported using marijuana within the past year. Rates of use among the colleges ranged from zero per cent at the lowest use schools to 54% at the highest use schools. Multiple regression models, constructed to determine the college and student characteristics predicting marijuana use, suggest that use was higher among students at non-commuter colleges and at colleges with pubs on campus. Student characteristics associated with marijuana use included being single, white, spending more time at parties and socializing with friends, and less time studying. Marijuana use was higher among students who participate in other high risk behaviors such as binge drinking, cigarette smoking and having multiple sexual partners, and among students who perceived parties as important, and religion and community service as not important. The study points to the social nature of drug use in college, and demonstrates that this behavior is of continuing concern for public health. PMID- 9219380 TI - Adulthood functioning: the joint effects of parental alcoholism, gender and childhood socio-economic stress. AB - The popular literature has publicized the adjustment difficulties of adult children of an alcohol-dependent parent (ACOAs); however, empirical studies do not provide consistent support. We examined the impact of parental alcoholism, degree of childhood socio-economic stress and gender on three broad categories of adulthood functioning (psychopathology, socio-economic attainment and marital stability). These effects were investigated with a heterogeneous sample of 400 men and 226 women participating in studies at the University of Michigan Alcohol Research Center. Parental alcoholism and childhood socio-economic stress exerted significant independent effects on most adulthood functioning measures. Men and women differed substantially only on socio-economic attainment measures, and effects of parental alcoholism and childhood economic stress on men and women were generally similar. For marital stability, parental alcoholism and childhood socio-economic stress interacted. These results suggest that researchers who study the impact of family history for alcoholism on psychological functioning should consider other aspects of the family of origin that promote wellbeing. In addition, results of this study point to the need for more research on gender differences, protective factors that promote good adjustment and outcome measures reflecting general life adaptation. PMID- 9219381 TI - The effects of physical attractiveness on gaining access to alcohol: when social policy meets social decision making. AB - Despite numerous legal interventions, minors continue to purchase and consume alcohol. Prior research had suggested that the decision to request identification to prove legal age was susceptible to various judgement and decision heuristics. This research examined whether the physical attractiveness of the potential consumer and the presence or absence of others were significant predictors of alcohol accessibility. Bartenders (n = 130) rated a target individual who was either high or low in attractiveness. Results indicated that attractiveness was a significant predictor of "proofing likelihood". High levels of attractiveness were associated with a decrease in the likelihood of being asked to provide proof of legal age for the purchase of alcohol. Individuals presented alone were seen as significantly older than when grouped with others. Implications of these findings for the restriction of alcohol availability among minors are considered. PMID- 9219382 TI - Are reconstructed self-reports of drinking reliable? AB - When follow-up interviews are missed, researchers sometimes try to reconstruct the data that would have been obtained by asking clients to recall the missed interval when they are interviewed at a later point. Are such data reliable? The reliability of remote reconstruction was estimated by asking 57 participants in a clinical trial to recall their drinking for the 12-month follow-up interval when interviewed, on average, 33 weeks later. These reports were obtained after delays averaging 231 days. These reconstructed reports were compared with the same clients' self-reports obtained during the 12-month interview. Reconstructed data were found to be reasonably accurate estimates of clients' reports at the time of original interview on global alcohol use variables including percentage of drinking days and total volume of consumption. No systematic bias was found for over-reporting or under-reporting at the point of reconstruction. However, on some variables (e.g. total drinks consumed), clients on average reported more drinking at the reconstruction period than during the initial interview. Discrepancies between initial and reconstructed reports were found to be unrelated to the length of delay in the second interview or to client characteristics. PMID- 9219383 TI - Do work-place smoking bans cause smokers to smoke "harder"? Results from a naturalistic observational study. AB - The purpose of this study was to investigate whether smokers outside buildings with work-place smoking bans smoke "harder" than those smoking in social settings. An unobtrusive random observational study of smokers followed by structured interview was used, with 143 smokers taking smoking breaks outside their office buildings and 113 smokers in social settings. The main outcome measurements were number of puffs per cigarette and cigarette smoking duration. The mean number of puffs per cigarette for the office building group was 18.7% greater than that for the social settings group (10.7 +/- 3.2 vs. 8.7 +/- 2.7, t = 5.58, df = 253, p < 0.001); 74.8% of smokers outside offices took more than the mean number of puffs for the group compared to 42.5% of smokers in social settings (chi 2 df 1 = 26.31, p < 0.0001). Mean cigarette smoking duration was 30.4% shorter for the work-place group than the social settings group (3.9 +/- 1.2 minutes vs. 5.6 +/- 2.6 minutes). Of smokers outside offices, 55.2% had a cigarette smoking duration between 3 and 4.59 minutes, while 53.1% of smokers in social settings took > or = 5 minutes to smoke the observed cigarette (chi 2 df 2 = 31.55, p < 0.0001). Smokers who scored at the 75th percentile on the Fagerstrom Tolerance Scale took a mean 9.5 +/- 2.6 puffs per cigarette compared to 9.3 +/- 2.7 puffs by those who scored in the 25th percentile on the scale (t = 0.34, df = 145, p = 0.73). Regardless of degree of nicotine dependency, smokers leaving work stations to smoke outside buildings smoked their cigarettes nearly 19% "harder" than cigarettes smoked in social settings. The individual and public health benefits of reduced smoking frequency engendered by work-place smoking bans may be lessened by policies which allow smokers to take smoking breaks. PMID- 9219386 TI - Society for Research on Nicotine and Tobacco. AB - The proceedings of the second annual scientific conference of the Society for Research on Nicotine and Tobacco are summarized. The goal of the annual conference was to disseminate information about ongoing nicotine research from biological, behavioral and social perspectives. Data were presented describing our current understanding of the structure and function of neuronal nicotinic acetylcholine receptors, by which nicotine exerts most, if not all, of its effects in the brain. The conformational complexity of receptor subunits expressed in different brain areas contributes significantly to the complexity of responses observed to nicotinic agonists. Nicotine is being developed as a medication that might be used to maintain smoking cessation and to treat various medical diseases. The potential toxicity of nicotine, apart from cigarette smoking, is an important variable in assessing the benefits and risks of such therapeutic applications. The risks of nicotine-containing medications appear to be far less than those associated with tobacco use. Recent data indicate that cigarette smoking is increasing among young in the United States. Adolescent smokers are interested in quitting and make frequent quit attempts, but are usually not successful. Effective methods are needed to manage adolescent smokers before they become heavily addicted. Nicotine replacement as a pharmacological treatment for smoking cessation has made a significant contribution in improving quit rates. New medications have been developed that target specific populations of smokers. PMID- 9219387 TI - Addiction history. PMID- 9219388 TI - The prepared mind: Marie Nyswander, methadone maintenance, and the metabolic theory of addiction. AB - Marie Nyswander was the co-discoverer of methadone maintenance. In the 1960s she and Vincent Dole carried out clinical trials and established the first officially sanctioned methadone clinics, which became models for maintenance programs throughout the world. In 1967 she and Dole theorized that heroin addicts had undergone a permanent metabolic change. They needed narcotics in a visceral way, which explained why abstinence was not a realistic goal-a claim that remains hotly disputed. Nyswander's work in the 1960s marked an abrupt shift in her thinking about addiction. Her previous approach had been centered on psychoanalysis. Her embrace of methadone maintenance and the metabolic theory amounted to an intellectual conversion. Nyswander was prepared for that conversion by years of patient relapse; she was frustrated with orthodox therapy. At the same time the success of such drugs as chlorpromazine suggested that psychiatric disorders might yield more readily to chemotherapy than talking therapy. Her own heavy smoking may have led her to question whether addiction was a psychiatric disorder in the first place. Finally, Nyswander had a long history of self-reinvention, having previously changed her politics, her medical specialty, and even her name. She was temperamentally suited for a conversion experience. PMID- 9219389 TI - Opium revisited: a brief review of its nature, composition, non-medical use and relative risks. AB - Unlike the pure opioids such as morphine and heroin, opium is a complex and variable mixture of substances reflecting differences in both the starting material and the traditional practices of the regions in which it is produced. Analytical methods have improved greatly in recent years, to the point that the source of a preparation can often be identified by its opioid content and its impurities. Daily amounts used, both by smoking and by mouth, vary widely from less than a gram to 30 g, equivalent to 75-3000 mg of morphine. The effects of opium are essentially those of morphine but unexpected toxicities, such as oesophageal cancer associated with "dross opium" and polyneuropathy due to deliberate addition of arsenic, are problems in some specific regions. Prevalence of use in different areas and countries is governed by the same factors of ease of availability, price and social acceptance that apply to the use of alcohol and other drugs in western countries. The risk of addiction to opium smoking appears to be somewhat less than to parenteral use of heroin, but appreciably greater than to alcohol. Even in countries where its use is traditional, opium smoking carries substantial risks of harm to health and social functioning. PMID- 9219390 TI - Early onset cannabis use and psychosocial adjustment in young adults. AB - The relationships between early onset (prior to 16 years) cannabis use and later psychosocial adjustment was examined in a birth cohort of New Zealand children studied to age 18 years. Early onset users had significantly higher rates of later substance use, juvenile offending, mental health problems, unemployment and school dropout. The linkages between early onset cannabis use and later outcomes were largely explained by two routes that linked cannabis use to later adjustment. First, those electing to use cannabis were a high risk population characterized by social disadvantage, childhood adversity, early onset behavioural difficulties and adverse peer affiliations. Secondly, early onset cannabis use was associated with subsequent affiliations with delinquent and substance using peers, moving away from home and dropping out of education with these factors in turn, being associated with increased psychosocial risk. The implications of these results are examined. PMID- 9219391 TI - Measurement properties of quantitative urine benzoylecgonine in clinical trials research. AB - Psychometric data are presented which examine the validity of using the concentration of benzoylecgonine in urine, a major metabolite of cocaine, as a measure of drug use, in studies of drug abuse treatments. In such research the standard biological indicator of drug use is usually a qualitative urine drug test, which merely indicates the presence or absence of a drug or its metabolite. A quantitative (i.e. continuous) outcome measure, such as the concentration of a drug or its metabolite in a biological fluid, has substantially more statistical power than a dichotomous measure and should, therefore, prove a more sensitive measure of drug use when viewed from a measurement perspective. Data from two placebo-controlled clinical trials of fluoxetine as an adjunct to treatment for cocaine abuse are analyzed to address this issue. Results indicate that urine benzoylecgonine level is closely related to self-reports of drug use and is independent of levels of anxiety, depression and hopelessness. PMID- 9219392 TI - Solitary drinking: a risk factor for alcohol-related problems? AB - This paper investigates whether solitary drinking is a risk factor for alcohol related problems using data from a general population of drinkers in Montreal, Canada. Three indicators of solitary drinking were used: (1) having had a drink alone; (2) frequency of solitary drinking; and (3) having had five drinks or more in a solitary setting. Among the 2015 respondent drinkers of a telephone survey, 31% reported drinking alone, of whom 27% did so more than once a week, and 17% had had five drinks or more alone at least once. Problems with family or social relationships, physical health, work, budget, physical security and happiness or view of life, self-reported as being alcohol-related, were measured by seven binary items. Strong positive associations were found at the univariate level between overall alcohol-related problems and both solitary drinking and having had five or more drinks alone, whereas frequency of solitary drinking had no effect. Only the relationship with having five or more drinks alone remained statistically significant in logistic regressions controlling for potential confounders. No evidence was found that solitary drinking per se is a risk factor for alcohol-related problems unless large quantities of alcohol are involved. PMID- 9219393 TI - End-of-treatment self-efficacy: a predictor of abstinence. AB - Results of previous studies suggest that end-of-treatment self-efficacy in problem drinkers has limited predictive validity. One explanation for this finding has been the postulated existence of a ceiling effect, i.e. the possibility that subjects who rate themselves highly in terms of self-efficacy form a heterogeneous group with some subjects making inflated self-efficacy judgements based on an over-optimistic perception of their coping abilities. In the present study, end-of-treatment self-efficacy in 63 problem drinkers, as measured by the Situational Confidence Questionnaire and a newly designed Self Efficacy Questionnaire (SEQ), was predictive of abstinence status at 3 month follow-up. In those patients who on the SEQ had expressed great confidence in their ability to remain abstinent over the follow-up period, the additional consideration of keyworkers' confidence in their patients' ability to remain abstinent as well as patients' anticipated need for future help improved the prediction of abstinence status. These results are discussed with respect to the postulated ceiling effect. A prognostic tree using just three baseline variables predicted abstinence status correctly in 88% of all cases. PMID- 9219394 TI - The Swiss Hidden Population Study: practical and methodological aspects of data collection by privileged access interviewers. AB - In order to recruit heroin and/or cocaine users outside treatment settings, recruitment of subjects through Privileged Access Interviewers (PAI) was tested and implemented in the Swiss Hidden Population Study. This article discusses practical aspects of the PAI method as well as issues of reliability and validity. From June 1994 to June 1995, 31 Privileged Access Interviewers were recruited in the main regions of Switzerland. They conducted 943 standardized interviews altogether, of which 917 could be considered valid. Fifty-four per cent of the respondents correspond to the criteria of the target population. The PAI method collects reliable data in a relatively short amount of time, given adequate means of control. Analysis of the age distribution and of the patterns of drug use in our sample shows that the question of validity is mainly linked to the diversity of the milieus from which PAIs recruit the respondents. Encouraging PAIs to do as many interviews as possible did not skew the data. Hence, well founded inferences on a PAI generated database relies on the analysis of qualitative information on the ways in which the Privileged Access Interviewers have recruited their respondents. PMID- 9219395 TI - The Dutch instant lottery: prevalence and correlates of at-risk playing. AB - After a long and contentious political debate, the instant lottery was introduced in the Netherlands in 1994. One of the conditions for allowing the introduction was that an evaluation study should be conducted with regard to possible negative side effects of the instant lottery in terms of excessive playing or addiction. This article reports on the main results of this evaluation study. In a random sample of 4497 instant lottery players, at-risk players were differentiated from recreative players on the basis of level of involvement in the instant lottery, impaired control and the experienced negative consequences of playing. Of the sample, 4.1% could be classified as an at-risk player. Actual problems resulting from playing in the instant lottery were experienced by 0.7% of the players. At risk players and recreative players did not only differ substantially in their playing behaviour, but also with regard to their socio-economic background, playing motivation, participation in other games of chance, and involvement in alcohol use and use of marijuana. To summarize, at-risk players were more likely to come from a poor socio-economic background, to play the instant lottery with a negative playing motivation, to be heavily involved in other forms of gambling, to have used marijuana and to drink alcohol excessively. PMID- 9219397 TI - Test-retest reliability of the Severity of Dependence Scale. PMID- 9219396 TI - Saliva cotinine levels as a function of collection method. AB - Saliva cotinine is commonly used to estimate nicotine intake but laboratories use different methods of collection. In three small trials, comparisons were made between (1) sugar vs. unstimulated saliva production (n = 29), (2) wax chewing vs. unstimulated production (n = 15) and (3) between two consecutive unstimulated saliva samples (n = 10). Sugar-stimulated saliva cotinine scores were 26% below unstimulated levels (p < 0.001); correlation between measures was high (r = 0.90; p < 0.001). Wax stimulated saliva yielded levels 6% below unstimulated (p < 0.05; correlation: r = 0.98; p < 0.001). No differences were observed between two unstimulated samples taken within a approximately 20-minute period (correlation: r = 0.99; p < 0.001). It is postulated that changes in salivary flow can account for the findings. PMID- 9219398 TI - Comment on Hall & Dolan's editorial on contact tracing. PMID- 9219399 TI - Influenza vaccine and older people: an evidence-based policy? PMID- 9219400 TI - Exercise prescription in primary care. PMID- 9219401 TI - Guidelines for dyspepsia management in general practice using focus groups. AB - BACKGROUND: There is a paucity of published guidelines on managing dyspepsia in general practice. Existing guidelines emphasize the role of investigations and drugs rather than management approaches. Focus groups are a means of uncovering the way in which the participants think and work in the pragmatic-setting, and have not previously been formally used in creating guidelines. AIM: To develop guidelines for the management of dyspepsia and to assess the use of focus groups of general practitioners (GPs) in order to do so. METHOD: Initial evidence-based guidelines were proposed by a group of four GPs with an audit facilitator, and used for discussion in three focus groups using a standard format. An anthropological analysis of the proceedings led to modifications of the original guidelines, based on knowledge, perceptions and attitudes. The study was set in three distinct locations involving 30 GPs. The outcome measures consisted of feedback, categorized by types of responses, from the analysis of the focus groups and the creation of guidelines. RESULTS: The resulting guidelines were patient centred and based on the principles of good consultation. They encompassed patients' fears and doctors' clinical uncertainties, and allowed flexibility in the individual patient's management. The focus group methodology exposed a substantial number of GPs to guideline development, and had the added benefits of dissemination, peer review and educational challenge. CONCLUSION: It was possible to develop guidelines for dyspepsia using focus groups. The methodology had the added benefits of ownership, peer review, exposure of educational gaps and locality factors, and dissemination of good practice. It included steps from evidence review to implementation strategies. The development of this technique could lead to a strategy towards the creation and application of evidence-based and professionally acceptable clinical guidelines and practice on a locality basis nationally. PMID- 9219402 TI - A randomized controlled trial of antibiotics on symptom resolution in patients presenting to their general practitioner with a sore throat. AB - BACKGROUND: Sore throat is a common symptom presented to general practitioners (GPs), and there remains controversy about the appropriate use of antibiotics. AIM: To compare, in a randomized controlled trial, the effectiveness of penicillin, cefixime and placebo on symptom resolution in patients presenting with a sore throat in general practice. METHOD: Twenty-two GPs in Avon recruited 154 patients, aged 16-60 years, presenting to their GP with a sore throat, and for whom the GP would normally prescribe an antibiotic. Patients were randomized to one of three groups: penicillin V 250 mg four times a day; cefixime 200 mg daily; and placebo. Each was prescribed for five days. The main outcome measures were a diary of symptom resolution over seven days and eradication of group A beta-haemolytic streptococcus (GABHS). RESULTS: Of the 103 (67%) patients who completed symptom diaries, 40 were allocated to receive penicillin, 29 cefixime and 34 placebo. In the analysis including all patients, symptom resolution was greater by day 3 in the cefixime group than in the placebo group. Penicillin did not improve symptom resolution by day 3 compared with placebo, and cefixime was not statistically significantly different from penicillin. There were significant differences in the proportion of patients using analgesia at day 3, with the proportion being lowest in the cefixime group. The results for the subgroup of patients without GABHS were similar to those for all patients; in particular, the only statistically significant difference was between cefixime and placebo. Although numbers were too small for statistical significance, among patients with GABHS the effects of penicillin and cefixime were similarly raised in relation to placebo. CONCLUSION: Compared with placebo, cefixime can improve the rate of resolution of symptoms in patients with a sore throat who are selected for antibiotic treatment by their GP. The unexpected finding that cefixime was of benefit compared with placebo for patients without GABHS suggests that bacteria other than GABHS may be important in the pathogenesis of sore throat. PMID- 9219404 TI - Training for audit: lessons still to be learned. AB - BACKGROUND: Audit is a criterion for training in general practice, and registrars are reliant on their trainers' teaching of basic audit methods. Their ability to teach this had been assumed, but registrars' projects submitted as part of summative assessment offered an opportunity to test this. AIM: To test trainers' knowledge of basic audit methods. Their knowledge was based on an ability to recognize key audit criteria using a marking schedule that they had helped to create. METHOD: All 158 trainers in the west of Scotland were asked to mark five general practice registrar audit projects using a marking schedule consisting of five independent criteria. Each project had one criterion that was below a level of minimum competence, as agreed by a group of 'expert' assessors. RESULTS: A total of 114 trainers (72%) completed the marking exercise of five audit projects. Three (3%) correctly identified the five criteria that were below minimum competence. They did this by highlighting many other criteria not below minimum competence. For all trainers, there was a direct relationship between the number of criteria they correctly identified as being below minimum competence and the total number of other criteria that they incorrectly identified. CONCLUSION: Trainers are failing to recognize basic audit methodology using a marking schedule they themselves helped to design. This has implications for their ability to teach audit to their registrars and may explain some of the difficulty in implementing audit. PMID- 9219405 TI - Diagnostic ultrasound: a primary care-led service? AB - BACKGROUND: A training programme has been proposed for general practitioners (GPs) to perform ultrasound in primary care. This has generated considerable concern among radiologists as to the adequacy and appropriateness of such training. AIM: To assess the current provision of ultrasound services to primary care in the former Northern health region of England, the level of interest among GPs in undertaking recommended training, and the willingness or ability of radiology departments to provide it. METHOD: Postal questionnaires were sent to GPs (n = 334), their practice managers (n = 281) and all clinical directors/heads of radiology departments (n = 19) in the region. RESULTS: Altogether, 67% of GPs, 59% of practice managers, and 68% of radiologists returned questionnaires. Overall, 48% of GPs have open access to obstetric/gynaecological ultrasound compared with 77% for general diagnostic requests. A total of 73% of GPs would prefer an open access service and 15% a practice-based service. Some 48% of GPs were not interested, 36% moderately interested, and 16% very interested in participating in the training programme. Only two out of 13 radiology, departments were willing to provide such training. CONCLUSION: Despite recommendations from the Royal College of General Practitioners, around half the respondents in this survey do not have direct access to ultrasound for obstetric referrals, and a quarter for non-obstetric referrals. Interest shown by GPs in a primary care-led service is not mirrored by their radiology colleagues. Open access to ultrasound was considered the optimum service, suggesting that resources be targeted at improving hospital services rather than transferring facilities to primary care. PMID- 9219403 TI - A survey of atrial fibrillation in general practice: the West Birmingham Atrial Fibrillation Project. AB - BACKGROUND: The management of atrial fibrillation (AF) has changed substantially in recent years, especially with a greater appreciation of the prophylactic role of antithrombotic therapy against stroke. There is therefore a need for further information on the prevalence of AF in Britain, the prevalence of (and contraindications to) anticoagulant treatment, and the factors that influence doctors' decisions in treating AF, including the investigation of patients with this arrhythmia. AIM: To investigate the prevalence, clinical features and management of patients with AF in a general practice setting. METHOD: Cross sectional survey of patients using treatment prescriptions and clinical records in two general practices from the west of Birmingham (serving a patient population of 16,519) where 4522 subjects (27.4%) were aged > or = 50 years. RESULTS: One hundred and eleven (2.4%) patients who were aged > or = 50 years were found to be in AF (42 males; mean age 76.6, SD 9.1); 77.5% were Caucasian, 2.7% Afro-Caribbean, 0.9% Asian, and 0.9% mixed race; in 20 cases there was no information on ethnicity. Of the AF patients, 5.4% were aged 50-60 years, 16.2% aged 61-70 years, 20.7% aged 71-75 years, 20.7% aged 76-80 years, 24.3% aged 81 85 years, and 12.6% aged > 85 years old, with female patients being significantly older than males. Eighty-one patients (73%) had chronic AF, while 30 patients (27%) had paroxysmal AF. The most common associated factors were hypertension (36.9%) and ischaemic heart disease (28.8%), with no obvious cause for AF in six patients. Cardiac failure was associated with AF in 34 patients (30.6%), and stroke had occurred in 29 patients (18%). Only 20 patients (18%) had had an echocardiogram, 26 (23.4%) a chest X-ray, and 58 (52.3%) thyroid function test. Only 30.6% had ever presented to hospital practice. Warfarin was prescribed to 40 patients (36%), with anticoagulation intensity monitoring by the general practitioner (GP) in three cases (7.5%), by a hospital clinic in 30 (75%), and by both GP and hospital in seven cases (17.5%). Of those not anticoagulated (n = 71), only 12 patients (16.9%) had contraindications to warfarin therapy. Patients treated with warfarin were younger than those who were not prescribed warfarin (71.3 versus 79.6 years, P < 0.001). Aspirin was being prescribed for 21 patients (18.9%), primarily for previous myocardial infarction. Only five patients (4.5%) had ever had attempted cardioversion. CONCLUSION: Atrial fibrillation is a common arrhythmia in general practice, and is commonly associated with hypertension, ischaemic heart disease and heart failure. There is a suboptimal application of standard investigations and use of antithrombotic therapy or attempted cardioversion; and few patients have presented to hospital practice. Guidelines on the management of this common arrhythmia in general practice are required. PMID- 9219406 TI - How general practitioners manage children with urinary tract infection: an audit in the former Northern Region. AB - BACKGROUND: Urinary tract infections (UTIs) in childhood are common and may be difficult to diagnose because of non-specific symptoms and technical problems with urine collection. Active management is important because UTIs may cause permanent renal scarring in young children. AIM: To determine how general practitioners (GPs) manage children with suspected UTIs. METHOD: A postal questionnaire to 494 GPs in the former Northern Region (a random selection of 26.2%) asking how they manage children with suspected UTI and their perception of their training needs. RESULTS: A total of 333 (67.4%) GPs replied. On weekdays, up to 22.9% of GPs treated children who had symptoms suggestive of UTI without collecting a diagnostic urine sample, and up to 64.8% did so at weekends. Urine collection was satisfactory in 73.2% of boys and girls aged under one year, but in only 50.4% of older boys and 48.0% of older girls, caused in part by the use of unreliably 'cleaned' potties in the older group. On weekdays, up to 87.2% of GPs culture the urine, but up to 4.8% use dipsticks as the sole diagnostic test; at weekends, only up to 58.6% culture urines, and up to 19.1% rely on dipsticks alone. Up to 11.0% of GPs examine urine under a microscope for bacteria to test for UTI on weekdays and at weekends. Up to 23.8% of GPs who collect urines on weekdays wait for a positive culture result before starting antibiotics. At weekends, only 3.9% of GPs build in this delay to treatment, mainly because far fewer take urine samples at all. GPs refer younger children for diagnostic imaging more readily than older ones, and boys more readily than girls at all ages. Although virtually all GPs refer all children under five years, some still do so only after recurrent infections. Over half the GPs wanted more training in managing UTI in children. CONCLUSION: There is a wide variation in clinical practice by GPs. Some always appropriately collect and test urine samples, treat without delay and refer for imaging after one proven UTI. Some never collect urines, treat blindly and refer only young infants with recurrent UTIs. Many vary their standards of practice from weekdays to weekends. The provision for GPs of clear, local, practical guidelines, drawn up between paediatricians and GPs and backed up with study days, might produce a consistent improvement in standards. PMID- 9219407 TI - One-to-one teaching with pictures--flashcard health education for British Asians with diabetes. AB - BACKGROUND: Type 2 diabetes is up to four times more common in British Asians, but they know little about its management and complications. AIM: To design and evaluate a structured pictorial teaching programme for Pakistani Moslem patients in Manchester with type 2 diabetes. METHOD: A randomized controlled trial of pictorial flashcard one-to-one education in 201 patients attending a hospital outpatient clinic or diabetic clinics in ten general practices in Manchester. Patients' knowledge, self-caring skills and attitudes to diabetes were measured on four topics before the structured teaching, and compared with results six months later. RESULTS: All parameters of knowledge were increased in the study group; for example, percentage scores for correctly identifying different food values increased from 57% to 71% (Analysis of Variance (ANOVA) adjusted difference +11.8%) and knowledge of one diabetic complication from 18% to 78%. Self-caring behaviour improved, with 92% of patients doing regular glucose tests at six months compared with 63% at the start. Attitudinal views were more resistant to change, with patients still finding it hard to choose suitable foods at social occasions. Haemoglobin A1c control improved by 0.34% over six months (ANOVA adjusted difference, 95% CI -0.8% to +0.1%). CONCLUSION: It is concluded that this health education programme can empower Asian diabetics to take control of their diets, learn to monitor and interpret glucose results, and understand the implications of poor glycaemic control for diabetic complications. PMID- 9219408 TI - Clinical aspects of recurrent postpartum thyroiditis. AB - BACKGROUND: Postpartum thyroiditis (PPT), characterized by transient hyperthyroidism and transient hypothyroidism, occurs in 5-9% of women. It is accompanied by the presence of circulating antithyroid peroxidase antibodies (TPOAb) which have been associated with an increase in depressive symptomatology compared with TPOAb-negative women. AIM: To assess the frequency and nature of the syndrome in patients studied in detail after more than one pregnancy, as there are only sparse data on recurrence of PPT. METHOD: Fifty-four patients were identified who had participated in at least two of three detailed postpartum studies of thyroid and psychiatric function during the past 12 years in the Caerphilly and Cardiff regions of South Wales. They included two women who had had three pregnancies. All patients had been followed monthly postpartum for at least six months, and 44 had been followed for 12 months. RESULTS: Of the 13 patients who developed PPT after their first pregnancy, nine had a recurrence of dysfunction after a further pregnancy and four remained TPOAb positive. Of the 24 women who were euthyroid anti-TPO positive after the first pregnancy, six developed thyroid dysfunction after a subsequent delivery, 14 remained antibody positive and euthyroid, while four underwent seroconversion and were antibody negative. The control group of 17 women were antibody negative after the first pregnancy; 16 remained negative after a further pregnancy and one became anti-TPO positive. The severity of PPT was slightly, but not significantly worse after the second recorded pregnancy (67% hypothyroid versus 44% hypothyroid). Neither the maximum anti-TPO titre following the first pregnancy, nor the rise in titre during this period were predictive of outcome after a subsequent pregnancy. Data from 26 women showed that recurrent depression was seen in 15.4%; a further six were depressed after the first pregnancy only, and two during a further postpartum period. CONCLUSION: There was a 70% chance of developing recurrent PPT after a first attack, and a 25% risk even in women who were only anti-TPO positive without thyroid dysfunction during the first postpartum period. The recurrence of postpartum depression was not related to thyroid function. Patients noted to have thyroid dysfunction or just to be euthyroid but anti-TPO positive after pregnancy should be assessed carefully after a subsequent pregnancy. PMID- 9219409 TI - Warfarin anticoagulation in primary care: a regional survey of present practice and clinicians' views. AB - The demand for anticoagulation services is rising. Warfarin anticoagulation has been shown to reduce the risk of stroke in patients with non-valvular atrial fibrillation by 68%. This raises issues about how services are best organized to initiate and monitor anticoagulation in this potentially large group of patients. We report the results of a regional postal survey undertaken to describe the views of general practitioners and consultants regarding warfarin anticoagulation in light of this potentially high increase in demand. PMID- 9219410 TI - Do general practitioners inform patients of the association between Helicobactor pylori infection and gastric cancer prior to determining serostatus? AB - The availability of non-invasive tests for the detection of Helicobactor pylori infection in primary care is increasing. However, the World Health Organization (WHO) has recently labelled H. pylori as a Class 1 carcinogen. We describe how frequently general practitioners (GPs) inform patients of this association prior to offering an H. pylori serology test, and discuss the possible consequences of withholding this information. PMID- 9219411 TI - Repeat radiographs in GP referrals to an orthopaedic clinic. AB - Repeat radiographs are expensive, both financially and in terms of cancer risks. This study looks at new referrals to an orthopaedic clinic, and examines the extent and some of the causes of the problem. PMID- 9219412 TI - Should general practitioners refer patients directly to physical therapists? AB - Several advantages have been claimed for general practitioners having direct access to physical therapy (defined as having a practice-based physical therapist or open access to a hospital-based physical therapist), and general practice fundholders are increasingly committing resources to ensure such services are available to their patients. This may lead to potential increases in costs as a larger total number of patients are treated owing to improved access and awareness of such services. A review of the available published literature found eight studies that compared two or more models of providing physical therapy services. Analysis of the studies revealed that there are several advantages for patients who are referred directly for physical therapy. The main advantages are significant reductions in waiting times, convenience, reduced costs for the patient and a lower cost per treated patient. There is also some evidence that the recovery time may be slightly better for patients who have direct access to a physical therapist. PMID- 9219413 TI - The Australian Quality Assurance and Continuing Education Program as a model for the reaccreditation of general practitioners in the United Kingdom. AB - A Quality Assurance and Continuing Education Program has been developed in Australian general practice over the past nine years. This effectively integrates audit and education within a coherent strategy for quality improvement. The programme fulfils many of the same aims as current proposals for reaccreditation in the United Kingdom (UK). This report describes the operation of the programme and an analysis of the effects of the scheme. A similar quality assurance strategy is proposed for the UK, which would address many of the criticisms of postgraduate education and may provide a realistic model for reaccreditation. PMID- 9219415 TI - Which adolescents attend the GP? PMID- 9219416 TI - Evidence-based approach to treating depression. PMID- 9219414 TI - The implications of teenage pregnancy and motherhood for primary health care: unresolved issues. AB - Teenage pregnancy and motherhood have implications for several different aspects of primary health care. First, the provision of health education and contraceptive services is obviously relevant to the prevention of unplanned teenage pregnancy. Secondly, appropriate obstetric care should be provided for teenagers, who are at high risk of developing complications in pregnancy and childbirth. Thirdly, and perhaps even more significantly, there is the implication of care required to deal with longer-term adverse health consequences associated with teenage pregnancy. In each of these areas, certain issues remain unresolved. This paper identifies key questions that remain unanswered, including the possibility of long-term adverse physical and psychological health consequences for teenage mothers and their children. The conclusion is that further research addressing these unresolved issues is necessary in order to inform health professionals and allow the implications for primary care to be assessed. PMID- 9219417 TI - Exercise on the NHS: a cost-effective exercise for older adults. PMID- 9219418 TI - Complementary medicine. PMID- 9219419 TI - Ethnic monitoring in general practice. PMID- 9219420 TI - Grandma knew which medicines were best. PMID- 9219421 TI - Corneal exposure in herpes zoster ophthalmicus. PMID- 9219422 TI - PSA excess in the differential diagnosis of prostate carcinoma. AB - OBJECTIVE: To evaluate the efficiency of PSA excess in distinguishing prostate cancer (PC) in its early stages from benign prostatic hypertrophy (BPH) and compare it with the efficiency of serum PSA. METHODS: A cross-sectional study was carried out on 27 patients with PC and 46 with BPH, whose serum PSA and prostatic volume were determined. PSA excess was calculated as the difference between serum PSA and predicted PSA according to the gland volume, calculating the latter as the prostatic volume multiplied by factor 0.3 ng/ml/g. RESULTS: PSA excess values were significantly higher in patients with PC, while serum PSA levels were not different between the two populations studied. Receiver operating curves (ROC plots) showed a higher diagnostic utility for PSA excess, with a maximum efficiency of 73% at a cut-off point of -13 ng/ml. The predictive value of a positive result is slightly higher for serum PSA, but PSA excess showed a predictive value of a negative result superior to that of PSA. PMID- 9219423 TI - Survival of adults with AIDS in the United Kingdom. AB - Accurate estimates of expected survival times and survival rates of AIDS patients are important both for estimating the prognosis of individuals and for monitoring the progress of the HIV/AIDS epidemic as new treatments are introduced. They are also needed for projecting future numbers of AIDS cases. Data on reported AIDS cases held at the PHLS AIDS Centre at the Communicable Disease Surveillance Centre and the Scottish Centre for Infection and Environmental Health confirmed the time, age, and reporting delay effects identified in earlier analyses of the United Kingdom AIDS database. The duration of survival after AIDS is diagnosed has improved since the epidemic began--median survival was 10.6 months in cases diagnosed before 1987 and has been at least 18.4 months in cases diagnosed each year since then. People who are diagnosed younger live longer--median survival fell from 21.6 months at age 15 to 29 to 12.6 months at age 45 or over. Delay in reporting AIDS cases adversely affects survival estimates for cases reported in recent years. Survival was longer in cases reported over a year after diagnosis of AIDS--23.7 months compared with 16.9 months in those reported less than a year after diagnosis. The experience of the hospital, measured by its cumulative AIDS caseload, was an important factor in the survival of men who have sex with men presenting with Kaposi's sarcoma alone or 'other' diagnoses--survival was shorter for cases reported from smaller centres. Men who have sex with men with Pneumocystis carinii pneumonia alone or other opportunistic infections alone who were known to be HIV positive before being diagnosed with AIDS had a shorter survival after being diagnosed than those who were unaware of their HIV infection. This supports the hypothesis that treatment for HIV infection and prophylaxis may extend the period before AIDS develops but reduce the period between developing AIDS and dying. PMID- 9219424 TI - Outbreak of small round structured virus gastroenteritis arose after kitchen assistant vomited. AB - A wedding reception at a North Yorkshire hotel was followed by an explosive outbreak of gastroenteritis. The attack rate among the 111 guests was 50% and vomiting was a predominant feature. The results of laboratory and epidemiological investigations were consistent with a common source outbreak of small round structured virus (SRSV) infection genotype II. The source of the outbreak was traced to a kitchen assistant who suddenly became ill on the eve of the reception and vomited into a sink used for preparing vegetables. The sink was cleaned with a chlorine based disinfectant and used the next morning to prepare a potato salad, subsequently identified as the vehicle of infection in a cohort study of guests (odds ratio 3.21; CI 1.78-5.78, p = 0.0001). No other food was associated with illness. The outbreak provides further supporting evidence of the importance of vomiting in the transmission of SRSV infection, highlights the virulence of this group of viruses, and indicates their relative resistance to environmental disinfection and decontamination. It also highlights the need for the adequate training of catering staff and the implementation and enforcement of food hygiene regulations. PMID- 9219425 TI - Increased incidence of parvovirus B19. PMID- 9219426 TI - AIDS and HIV infection in the United Kingdom: monthly report. PMID- 9219427 TI - Molecular dynamics simulation of hydrated phospholipid bilayers. AB - Understanding of microscopic behaviour of biological membrane is crucial for designing of molecules to control transport properties of the membranes. Phospholipid-water forms a good model system to study ligand induced structural and dynamical changes in membrane. The review has its main focus on molecular dynamics (MD) simulation of phospholipid bilayers. A brief summary of the current status of structure of phospholipid membranes based on different physico-chemical measurements is given. We discuss here mainly results of MD simulations in the recent years on hydrated phospholipid bilayers and their interaction with ligands. Simulation parameters as: choice of initial system, force fields, protocols for simulation are compared. Main results on: order parameter, head group and chain conformation, water penetration profile, chain tilts, pair correlation function between atoms of lipid and water, diffusion of ions and ligands are discussed. The review gives application and limitation of MD method for studying lipid water system. PMID- 9219428 TI - Cucumber chloroplast trnL(CAA) gene: nucleotide sequence and in vivo expression analysis in etiolated cucumber seedlings treated with benzyladenine and light. AB - The nucleotide sequence of a 714 bp BamHI-EcoRI fragment of cucumber chloroplast DNA was determined. The fragment contained a gene for tRNA(Leu) together with its flanking regions. The trnL(CAA) gene sequence is about 99% in similarity to broad bean, cauliflower, maize, spinach and tobacco corresponding genes. The relative expression level of the gene was determined by Northern (tRNA) gel blot and Northern (total cellular RNA) slot-blot analyses using the trnL gene probe in 6 day old etiolated cucumber seedlings and the seedlings that had been kept in the dark (dark-grown), treated with benzyladenine (BA) and kept in the dark (BA treated dark-grown), illuminated (light-grown), and treated with BA and illuminated (BA-treated light-grown), for additional 4, 8 or 12 hr. The trnL transcripts and tRNA(Leu) levels in BA-treated dark-grown seedlings were 5 and 3 times higher, respectively after 4 hr BA treatment, while in the BA treated light grown seedlings the level of trnL transcripts was only 3 times higher and had no detectable effect on mature tRNA(Leu) when compared to the time-4 hr dark-grown seedlings. However, the level of mature tRNA(Leu) did not show marked changes in the light-grown seedlings, whereas the level of trnL transcripts increases 3 times after 8 hr illumination of dark-grown seedlings. These data indicate that both light and cytokinin can signal changes in plastid tRNA gene expression. The possible regulatory mechanisms for such changes are discussed. PMID- 9219429 TI - Alterations in brain tumor DNA detected by a fingerprinting probe. AB - We used a novel DNA fingerprinting probe O-chi-1 (ref. 1) to detect differences in the hybridization pattern of brain tumor DNA and paired normal tissue of a given individual. Representatives of meningiomas (two), glioblastoma multeforme (three) and astrocytoma (one) were studied. Alterations, which included amplification as well as the loss of a normal band in tumor DNA, were observed in four of the six tumours. While the increased intensity of a band can be taken to imply increased copy number, the disappearance of bands could either be due to loss of DNA sequence or rearrangement resulting in different sized bands. PMID- 9219430 TI - Molecular electrostatic potential mapping and structure-activity relationship for 3-methoxy flavones. AB - Molecular electrostatic potential (MEP) maps of certain 3-methoxy flavone derivatives having different anti-picornavirus activities have been studied. Geometries of the molecules were optimised and charge distributions computed using the AM1 molecular orbital method. Hybridization displacement charges (HDC) were combined with the Lowdin charge distributions to compute the MEP maps. Reliability of the method of computing MEP maps was tested by studying certain other molecules for which ab initio MEP results are available. The anti picornavirus activities of the flavones have been shown to be related with negative MEP values in two regions, one near the 3-methoxy group and another in a diagonally opposite region near the substituent attached to the C7 atom of the molecules. PMID- 9219431 TI - Studies on electron donation to photosystem I sites by exogenous donors in Spirulina thylakoids. AB - The kinetic parameters of different sites of electron donation to photosystem I (PS I) were evaluated in Spirulina platensis thylakoids. Reduced 2,6 dichlorophenolindophenol (DCIPH2) exhibited two sites of electron donation, with apparent K(m) values of 8 and 40 microM each. The corresponding value for reduced N-tetramethyl-p-phenylenediamine (TMPDH2) and diaminodurene (DADH2) which donate electrons at a single site to PS I were 103 and 48 microM, respectively. The electron donation by these three exogenous donors were differentially inhibited by KCN (70 mM) affecting the apparent K(m) and Rmax values to varying extent. This cyanide inhibition of PS I catalyzed electron transport suggests the presence of plastocyanin in the photosynthetic electron transport chain of Spirulina platensis. PMID- 9219432 TI - Effect of photoinhibition and temperature on carotenoids in sorghum leaves. AB - Changes in carotenoid composition, CO2 assimilation and chlorophyll fluorescence due to photoinhibition at 5 degrees C and 20 degrees C were studied in 12 day and 30 day old sorghum leaves. The old leaves had a higher violaxanthin (V) content and less beta-carotene. Photoinhibition at both temperatures caused significant increases in zeaxanthin (Z) and decreases in violaxanthin. However, in young leaves the increase in zeaxanthin was greater than the decrease in violaxanthin. In young leaves the V + A + Z pool size (A = antheraxanthin) almost doubled under photoinhibitory conditions (compared to controls) while in old leaves the V + A + Z pool remained approximately constant. After photoinhibition treatment changes in the levels of the xanthophylls were restored during a recovery period both in young and old leaves. When rephotoinhibited after a 48 hr recovery period, the young plants showed better protection against photoinhibition. We suggest that in young leaves zeaxanthin is newly synthesized under photoinhibitory conditions besides being de-epoxidized from violaxanthin and that the synthesis of V + A + Z pool is higher at 20 degrees C than at 5 degrees C in both young and old leaves. PMID- 9219433 TI - Fatty acid synthesis by isolated leucoplasts from developing Brassica seeds: role of glycolytic intermediates as the source of carbon and energy. AB - Fatty acid synthesis from Na [1-14C] acetate in leucoplasts isolated from developing seeds of Brassica campestris was completely dependent on exogenous supply of ATP. None of the intermediates of glycolysis or pentose phosphate pathway tested could replace ATP in the reaction mixture. In absence of exogenously supplied ATP, maximum activity was obtained with glu-6-P (68%) followed by fru-6-P (50%) and PEP (44%), respectively. With other intermediates as energy sources, the activity ranged from 1 to 38%. In complementary experiments (presence of ATP), none of the metabolites gave activity higher than the ATP control activity. Under optimum conditions for fatty acid synthesis from acetate, Brassica leucoplasts readily utilized labelled glucose as the substrate for fatty acid synthesis. Omission of NADH and NADPH individually from the reaction mixtures containing labelled glucose resulted only in 46 and 20% loss in activity, respectively, compared to the corresponding losses of 56 and 50%, when labelled acetate was used as the substrate. Similarly, deletion of ATP from the reaction mixture containing glucose as the substrate decreased the rate of fatty acid synthesis by about 65%, while the corresponding decrease with acetate as the substrate was 96%. Inclusion of 5 mM cold acetate, pyruvate, malate and glu-6-P in the reaction mixture containing glucose as the labelled substrate reduced label incorporation into fatty acids by 38 to 69%, maximum reduction being observed with pyruvate followed by glu-6-P, acetate and malate, respectively. With labelled acetate as the substrate, maximum reduction in label incorporation was obtained with cold glucose (5 mM) followed by glu-6-P, pyruvate and malate, respectively. The study demonstrated the operation of complete glycolytic pathway in Brassica leucoplasts, allowing the plastids to use glucose as a source of carbon, reducing power and energy for fatty acid synthesis. PMID- 9219434 TI - Possible role of calcium dependent protein phosphorylation in the modulation of wound induced HRGP gene activation in potatoes after gamma irradiation. AB - Hydroxyproline rich glycoprotein (HRGP) gene is induced in both control and gamma irradiated potato tubers after wounding. The enhanced RNA synthesis in response to wounding correlated well with the accumulation of both HRGP gene transcripts and protein. Initially, the level of HRGP gene expression in gamma irradiated potatoes in response to wounding was 30% more than the corresponding controls. After post irradiation storage of 3-5 weeks, HRGP gene expression in response to wounding was significantly lower than the unirradiated samples. This low level of HRGP gene expression in irradiated potatoes was partially retrieved by 5 mM Ca2+ treatment. Prior treatment with trifluoperazine, a calcium channel blocker resulted in 35% reduction in wound induced HRGP gene expression in control potatoes, further providing evidence for the involvement of Ca2+ dependency for HRGP gene activation. A comparative study on in vivo protein phosphorylation induced by wounding in control and irradiated potatoes exhibited significant differences. A good correlation was observed in the modulation of phosphorylation and HRGP gene expression by Ca2+ in irradiated potatoes. Wound induced signal transduction system and subsequent Ca2+ dependent protein phosphorylation for the activation of HRGP gene is affected in potatoes after gamma irradiation, thus impairing the wound healing process adversely. PMID- 9219435 TI - Serine proteinase from rice bean. AB - A trypsin like serine-proteinase of M(r) 16,000 Da, optimally active at pH 8.4 on N-benzoyl-arginine ethyl ester (BAEE) was purified from 4-day old germinated seeds of rice bean, Vigna umbellata (Thunb), by ammonium sulphate precipitation, gel filtration, ion-exchange chromatography and by high performance liquid chromatography (HPLC). The purity of the enzyme was checked by polyacrylamide gel electrophoresis (PAGE). The enzyme activity was studied on natural substrates like casein, haemoglobin and vicilin, a rice bean storage protein. The activity of the enzyme was completely inhibited by phenylmethylsulfonyl fluoride, but not by iodoacetamide and HgCl2, suggesting it to be a serine protease. Loss of activity in presence of EDTA was reversed by addition of Ca2+. PMID- 9219436 TI - Aggregation of banana pyrophosphate fructose 6-phosphate 1-phosphotransferase by glycerol. AB - Addition of glycerol during purification of banana (Musaceae, Musa cavendishii) pyrophosphate fructose 6-phosphate 1-phosphotransferase [(PFP), EC 2.7.1.90] initiated molecular aggregation of the enzyme. The aggregation process was dependent on the glycerol concentration. The native enzyme (66 kDa molecular mass) showed enhanced activity at 3% (V/V) or less of glycerol concentration. Glycerol concentration between 4 and 5% (V/V) affected a gradual and sequential aggregation of native form of the enzyme. These aggregated forms had molecular masses of 135, 200 and 270 kDa. The 135 and 200 kDa forms were stable for about 72 hrs and prolonged storage over 2 weeks resulted in the formation of the 270 kDa form. Concentration over 5% could reduce the time required for aggregation. Fru2.6 bis P activated the enzyme over ten fold, but did not help in the aggregation process. Studies on the role of glycerol on PFP specific activity suggested a difference in the activation process compared to that by Fru2.6bis P. Replacement of Hepes buffer by Tris increased the Fru2.6 bis P requirement for maximum activation by around 10 fold. Removal of glycerol from the buffer media resulted in almost complete inactivation of the enzyme. PMID- 9219437 TI - Characterization of a lectin from goat peripheral blood lymphocytes. AB - A D-glucose specific lectin was isolated from goat peripheral blood lymphocytes by affinity chromatography on N-acetyl D-glucosamine agarose gel. The fluorescence intensity of 4 methyl umbelliferyl D-glucose was quenched to about 62% on addition of the lectin. This lectin gave a single band corresponding to 112 kDa in SDS-PAGE irrespective of treatment with 2-mercaptoethanol. The molecular weight and the Stoke's radius of the lectin in the native conditions were found to be 114 kDa and 4.54 nm, respectively, as determined by gel filtration on Sephacryl S 500 column. The lectin was found to be a glycoprotein with 5.6% of neutral hexose content and 5.5% of sialic acid. The lectin agglutinated trypsinized rabbit erythrocytes and human type A erythrocytes. The hemagglutinating activity was dependent on the presence of divalent cations like Mn2+ and Ca2+. Optimum pH, ionic strength and temperature for rebinding of lectin to acid treated Sephadex G200 were found to be 7.5, 0.16 and 30-37 degrees C, respectively. PMID- 9219439 TI - Isatin (2,3-dioxoindole): a competitive inhibitor of Na(+)-dependent lysine uptake in rat instestine. AB - Isatin (2,3-dioxoindole) competitively inhibited (27-40%) Na(+)-dependent L lysine uptake in rat intestine. The value of Kt was increased from 3.04 mM in control to 5.88 mM in presence of 10 mM isatin. Effect of isatin on the Na(+) independent amino acid uptake was insignificant (12-18%). The inhibitory constant (Ki) was 2.8 mM under these conditions. The observed inhibition was unaffected by -SH group reacting agents. Isatin (1-10 mM) inhibited Na+, K(+)-ATPase activity in intestine in vitro, the maximum inhibition (66%) being at 10 mM isatin concentration. But the drug had no effect on enzyme activity under in vivo conditions. PMID- 9219440 TI - Aryl monooxygenase: a detoxifying enzyme from Candida pulcherrima MCMY2. AB - Characterization of partially purified aryl monooxygenase (1:14:14:1) from Candida pulcherrima MCMY2 was achieved using standard purification protocol. The molecular weight of the enzyme was 110 kDa and contained 3 subunits with pI 5.7, 7.4 and 7.6. The activity seemed to be related with pulcherrimin. The optimum pH and temperature for enzyme activity were 6.8 and 30 degrees C respectively. Activity was not substrate specific and Fe3+ FAD and NADH+ enhanced the activity substantially. PMID- 9219438 TI - Regulation and properties of purified glucose-6-phosphate dehydrogenase from rat brain. AB - Glucose-6-phosphate dehydrogenase from rat brain was purified 13,000 fold to a specific activity of 480 units/mg protein. The molecular weight was 121 kDa. The kinetics of brain glucose-6-phosphate dehydrogenase are compatible with a model involving two possible states of the enzyme with a low and high affinity for the substrate D-glucose-6-phosphate. NADP+ and ADP offered protection against p chloromercuribenzoate inhibition. NADPH is a powerful competitive inhibitor with respect to NADP+. The apparent Ki for NADPH inhibition was lower than the Km for NADP+. ADP inhibited the enzyme competitively with respect to NADP+. ATP inhibited the enzyme non-competitively with respect to NADP+, whereas kinetics of mixed inhibition was observed with respect to substrate D-glucose-6-phosphate. The interplay between NADP+ and NADPH leading to enzyme activation or inhibition according to their relative or absolute concentrations as well as the control of enzyme activity by the adenine nucleotide system may contribute a refined mechanism for the regulation of glucose-6-phosphate dehydrogenase and therefore the pentose phosphate pathway in brain. PMID- 9219442 TI - Changes in hydroxyproline levels in electric field tissue interaction. AB - Guinea pigs when exposed to external electric field (1.9 kV/m) for 9 hrs/day for 3 days registered rise in the levels of hydroxyproline in liver, lung and kidney, suggesting increased synthesis of collagen. PMID- 9219441 TI - Metabolism of proteins and glycoproteins in tumour bearing mice treated with Aeromonas L-asparaginase. AB - L-asparaginase, isolated in our laboratory, from Aeromonas had been found to be antileukaemic. In the present study, changes in the levels of proteins and glycoproteins in leukaemic mice and under treatment with Aeromonas L-asparaginase have been compared. Levels of protein bound hexose, fucose and sialic acid which were increased during leukaemia attained normal levels when treated with L asparaginase. The increased blood urea level declined significantly during enzyme therapy. Effects of L-asparaginase are compared with 'Leunase', a commercially available drug used in the treatment of leukaemia. PMID- 9219443 TI - Evaluation of Leishmania antigens preparation and storage for use in enzyme immunoassays. AB - The influence of time and temperature on the storage of an alkaline antigen of L. major-like and L.(V.) braziliensis promastigotes added or not of a proteases inhibitor (PMSF) was evaluated by means of an IgG-ELISA. Antibodies in assays using L. major-like antigen stored at -20 degrees C for 6 months had a statistically lower geometric mean titer (GMT) and different 95% confidence interval limits (CL) than antigens stored otherwise, as assessed by the "t" statistic. The PMSFL. major-like antigen after storage for 6 months at a temperature of 4 degrees C had the same GMT and 95% CL displayed at time zero as well as when storage for 4 and 6 months at -20 degrees C. Significant differences were not found when L.(V.) braziliensis antigens were stored at times and temperatures mentioned; the PMSF antigen stored for 2 months at -70 degrees C resulted in a lower serum GMT and 95% CL than any other, as assessed by the "t" statistic. Antigen performance did not show any statistical difference associated to the addition of PMSF within the same species; the largest difference between antigens was that between PMSF-L. (V.) braziliensis and L. major-like without PMSF. PMID- 9219444 TI - Acute Chagas' cardiopathy in a polar bear (Ursus maritimus) in Guadalajara, Mexico. AB - We report a 24-year-old female polar bear (Ursus maritimus) who contracted Chagas' infection at the Guadalajara Zoo, in Jalisco, Mexico, and died of acute Chagas' carditis 15 days later. The histopathological findings are described, as well as the presence of triatomids (Triatoma longipennis Usinger) infected with Trypanosoma cruzi collected within 5 meters from the place where the animal lived in the city of Guadalajara. PMID- 9219448 TI - Why are some children stunted at birth, and do they catch up with their peers in infancy? PMID- 9219449 TI - Intrauterine growth pattern by the tendency to repeat small-for-gestational-age births in successive pregnancies. AB - BACKGROUND: Fetuses of women who repeat small-for-gestational-age births in successive pregnancies may have a different intrauterine growth pattern than SGA birth of non-repeater mothers. Also repeated SGA births may grow differently depending on whether the tendency to repeat is due to some external factors such as cigarette smoking ("false repeaters") or due to genetic or intrinsic factors ("true repeaters"). MATERIAL AND METHODS: Fetal growth were compared in a "nested case-control" study within a longitudinal (cohort) study, comparing three types of SGA births, 23 of "true repeater" mothers, 46 of "false repeater" mothers and 65 of non-repeater mothers, and these were compared with 1017 non-SGA births. Fetal growth was compared using a regression analysis based on repeated measurements (four for each woman). RESULTS: For mean abdominal diameter the "true repeater" SGA births grew more slowly towards the end of pregnancy. However, the growth curves show only minor differences between the three types of SGA births, but the patterns are grossly different from the growth of non-SGA births (controls). CONCLUSION: The intrauterine growth retardation starts early in pregnancy, and is not strikingly different between births of repeater and non repeater mothers. PMID- 9219450 TI - The relationship between maternal characteristics and fetal and neonatal anthropometric measurements in women delivering at term: a summary. AB - BACKGROUND: We wanted to determine the relationship between a number of maternal characteristics and various fetal and neonatal anthropometric measurements determined by ultrasound and at birth. METHODS: A total of 1205 term singleton maternal-infant pairs were studied. Various ultrasound measurements obtained at 18, 24, 30 and 36 weeks' gestation and neonatal anthropometric measurements obtained at birth were studied in relationship to various maternal characteristics using univariate and multivariate techniques. RESULTS: Black race, female sex, cigarette smoking, drug use, having a previous low birthweight infant, maternal hypertension and being short or thin or failing to gain weight each resulted in a birthweight decrease of 100 to 300 g. The effect of each of these characteristics on each ultrasound measurement, the timing of the effect, and its ultimate effect on neonatal anthropometric measurements are described. CONCLUSION: The data presented in this paper provide a more complete understanding of the relationship between maternal characteristics, infant sex, and various fetal ultrasound and neonatal measurements. PMID- 9219451 TI - Psychosocial factors and small-for-gestational-age infants among parous Scandinavian women. AB - BACKGROUND: We wanted to analyze the association between small-for-gestational age (SGA) births, defined as a newborn with a birthweight below the 15th percentile-for-gestational age, and socioeconomic and psychosocial risk factors. METHODS: Information on social background, psychological status, and life events was collected prospectively by use of questionnaires in the second and third trimester of pregnancy. The respondents were 1552 women who expected their second or third child and took part in a Scandinavian multicenter study of fetal growth and perinatal outcome. RESULTS: No significant differences were found in relational stress, state and trait anxiety, depression, and physical strain between SGA and non-SGA births, whereas smoking around time of conception and low prepregnant body mass were significant SGA birth predictors. Maternal and paternal education of nine years or less increased the SGA birth risk (RR 1.46 (95% CL 1.12; 1.92) and RR 1.34 (95% CL 1.01; 1.79), respectively. The increased risk from a low maternal education was still significant when body mass and low paternal education were controlled, but not after adjustment for maternal smoking. A protective effect of paternal, but not maternal, education of 12 years or more was also observed and retained its effect when maternal smoking and body mass were controlled. CONCLUSION: In this seemingly homogeneous Scandinavian population, parental education and maternal body proportion and life style influenced the prevalence of small-for-gestational-age births. Relational stress, anxiety, depression, and physical strain did not influence birth outcome. PMID- 9219453 TI - Fetal growth impairment from smoking--is it influenced by maternal anthropometry? AB - BACKGROUND: It has been suggested that the effect of maternal smoking on fetal growth is partly mediated through nutritional factors. OBJECTIVE: To assess the effects of maternal smoking on birthweight in term pregnancies among mothers with different anthropometric stature. DESIGN: A prospective study from early pregnancy of healthy parous women and their infants. SETTING: Three Scandinavian university hospitals covering all deliveries from well defined geographic areas. SUBJECTS: Smoking (774) and non-smoking (325) mothers, para 1 and 2 and with > 36 weeks gestational length. MAIN OUTCOME MEASURE: Birthweight. RESULTS: Maternal age, smoking, pre-pregnancy weight, height, body mass index and pregnancy weight gain all independently influenced birthweight. Smoking mothers had significantly lower pre-pregnancy weight and lower body mass index compared to nonsmoking mothers. The negative influence of smoking on birthweight appeared to be uniformly distributed throughout all the different maternal height and weight groups. CONCLUSIONS: Birthweight was negatively related to amount cigarettes smoked per day, and no protective effect could be demonstrated from higher maternal weight, pregnancy weight gain or body mass index. PMID- 9219452 TI - Abbreviated scale for the assessment of psychosocial status in pregnancy: development and evaluation. AB - BACKGROUND: Data from five existing psychosocial scales were used to develop an abbreviated scale for the assessment of psychosocial status during pregnancy. METHODS: Scales were self-administered by 842 black and 381 white low-income multiparous women at risk for poor pregnancy outcome. Trait anxiety (Speilberger), self-esteem (Rosenberg), mastery (Pearlin), and depression (CES-D) were assessed at 24-26 weeks' gestation; subjective stress (Schar) was assessed at 30-32 weeks' gestation. The 59 pooled items were examined for redundancy and the discernment of primary factors using principal factor analysis. Regression analysis was used to determine if the resulting abbreviated scale (28 items) would provide information similar to that obtained with the 59 item pool (full scale) in predicting gestational age (GA), birth weight (BW), fetal growth restriction (FGR), and preterm delivery (PTD). RESULTS: The abbreviated scale was highly correlated (r = 0.97) with the 59-item pool and the six factors isolated were generally compatible with the major characteristics assessed by the five original scales. The distribution of FGR and PTD by scale quartile was similar for the abbreviated and the combined scales. Logistic regression analysis of scores for all women revealed that poor (high) scores on both the full (p = 0.0151) and the abbreviated scales (p = 0.0131) were positively associated with FGR, but not with PTD. In linear regression analysis poor (high) scores on both the full (p = 0.0024) and the abbreviated scale (p = 0.0019) were negatively related to BW, but not to GA. When data for black and white women were examined separately, the two scales provided comparable information. CONCLUSIONS: The abbreviated psychosocial scale provided information similar to that obtained with 59 pooled items in predicting GA, BW, FGR, and PTD. PMID- 9219454 TI - Birth weight of relatives by maternal tendency to repeat small-for-gestational age (SGA) births in successive pregnancies. AB - BACKGROUND AND METHOD: Small-for-gestational-age (SGA) infants represent a heterogeneous group of normal and growth-retarded children. To assess the familial aggregation of reduced fetal growth, birth weights in both maternal and paternal relatives of 1246 index children in the Scandinavian SGA Study were compared across groups defined by the SGA outcome of the index child as well as that of earlier siblings. RESULTS: Mean maternal birth weight +/- SEM was 3127 +/ 54 g for mothers who had experienced two SGA births as opposed to 3424 +/- 22 for mothers with no SGA births. Mean paternal birth weight was 3497 +/- 88 g and 3665 +/- 24 in the same two groups. The odds ratio (with 95% confidence interval) for having a mother with birth weight below the 10th percentile was 1.74 (0.85 3.58) for the group where two SGA births had occurred compared to no SGA births and it was 2.49 (1.22-5.07) for having a father with birth weight below the 10th percentile. There was no correlation between maternal and paternal birth weights. CONCLUSIONS: The association also to paternal birth weight suggests the presence of genetic or common environmental factors in explaining the tendency to have SGA children. Although taking parental birth weights into consideration will aid in diagnosing growth-retardation in a SGA child, SGA remains a heterogeneous group where familial and non-familial cases will be difficult to separate. PMID- 9219455 TI - Obstetric interventions and perinatal asphyxia in growth retarded term infants. AB - BACKGROUND: The monitoring of fetal growth during pregnancy is usually justified because of the increased perinatal risk of these babies. METHODS: In 1552 infants from the Scandinavian Small for Gestational Age Study the need for obstetric interventions, risk of fetal asphyxia and immediate neonatal outcome at term have been studied in relation to different types of fetal growth retardation, including sub-groups with low ponderal index or low amount of subcutaneous fat. RESULTS: The need for obstetric intervention indicated by suspected fetal asphyxia before or during labor was increased 3-fold (6-8%) for growth retarded infants both in SGA infants in general and infants with asymmetric body proportions. The immediate perinatal outcome, however, was favorable with Apgar below 8 at 5 min in only 2% irrespective of the type of growth retardation, in spite of the fact that less than 25% of the SGA pregnancies and 10% of those with asymmetric fetal growth had been eligible for close antenatal fetal monitoring. CONCLUSION: With a moderate increase in interventions at delivery, perinatal outcome was highly favorable for term infants with a weight for gestational age, weight for length or skinfold for weight below the 10th percentile in this population of Scandinavian parous mothers. PMID- 9219456 TI - Pre-pregnancy risk factors of small-for-gestational-age births and perinatal mortality. AB - The Scandinavian part of the NICHD study of successive small-for-gestational-age (SGA) births included 5722 parous women from Trondheim and Bergen (in Norway) and Uppsala (in Sweden). Study enrollment took place from January 1986 through March 1988. The aim of the main study was to investigate factors associated with inhibited intrauterine growth. This paper reports on the fetal, perinatal, and neonatal deaths among the births in the study in relation to different risk factors. The cause of deaths were analyzed to see if there were any associations with the risk factors. There was a total of 84 deaths, 65 of these were fetal, perinatal or neonatal deaths and included in this analysis. The remaining 19 are for different reasons excluded. Thirty-two (60%) of the autopsies regarded the high risk group who comprised only 42.4% of the total study population. The high risk group was selected using the following risk criteria: a previous low weight birth or perinatal death, maternal low weight (pre-pregnancy weight < 50 kg), the presence of a chronic maternal disease, and smoking at the time of conception. A significant association was found between perinatal mortality and the presence of one or more of the defined risk criteria (relative risk 2.0; 95% CI 1.2, 3.4). Asphyxia and related disorders was the most important single cause of death and was found to be associated with the maternal risk factors (RR 3.9; 95% CI 1.5, 9.8). A significant association was found between maternal risk factors and SGA autopsies (RR 3.9; 95% CI 1.7, 8.9). No association was found between asphyxia and SGA. It is concluded that women with risk factors based on complications in a previous pregnancy are more prone to stillbirths, perinatal, and neonatal deaths, and with asphyxia as the most prominent cause of death. PMID- 9219457 TI - Prediction of fetal growth based on maternal serum concentrations of human chorionic gonadotropin, human placental lactogen and estriol. AB - BACKGROUND: The purpose was to determine the usefulness of maternal serum concentrations of human chorionic gonadotropin (hCG), human placental lactogen (hPL) and estriol as predictors of fetal growth. METHOD: From a large cohort serum obtained serially at 17, 25, 33 and 37 weeks of gestation were analyzed for randomly selected pregnancies resulting in small for gestational age (SGA, n = 102) and non-SGA (n = 112) infants. RESULTS: There were no significant correlations between birthweight ratio (ratio of birthweight to mean weight for gestational age) and hCG, but between birthweight ratio on one hand and estriol for all stages of pregnancy (r = 0.19-0.38, p < 0.01 - p < 0.001) and hCL except at 33 weeks (r = 0.11-0.40, p ns-p < 0.001) on the other. There were statistically significant, but small median differences and substantial overlaps between the SGA and non-SGA infants for hCG at 17 and 37 weeks, for hPL at 17, 33 and 37 weeks, and for estriol at all the stages of pregnancy. The sensitivity and positive predictive value of low hormone concentrations (below the 10th percentile) in predicting the birth of an SGA infant were in the range of 6-26% and 17-39%, respectively. The corresponding specificity and prediction of a non SGA infant from normal levels were 91-93% and 85-88%. CONCLUSIONS: HPL and estriol, but not hCG concentrations, are positively related to the size of the fetus, but the relationships are too weak to be of predictive value in an unselected population. PMID- 9219458 TI - Maternal serum concentrations of human placental lactogen, estradiol and pregnancy specific beta 1-glycoprotein and fetal growth retardation. AB - BACKGROUND: To determine if maternal serum levels of human placental lactogen (hPL), estradiol, and pregnancy-specific beta 1-glycoprotein (SP1) measured at approximately 18 weeks' gestation were associated with fetal growth retardation (FGR) in infants delivered at or after 37 weeks. METHODS: Serum samples were obtained at a mean of 18 weeks' gestational age from 200 multiparous women with risk factors for FGR. Maternal serum concentrations of hPL, estradiol and SP1 were correlated with FGR. RESULTS: A total of 59 (29.5%) of the 200 infants were diagnosed postnatally with FGR. There were no significant differences in the prevalence of FGR among the lowest quartiles of estradiol, hPL or SP1. However, pregnancies in the highest quartile of estradiol levels at 18 weeks' (> 580 pg/ml) were associated with a significantly lower risk of FGR than those in the lower three quartiles, 8 out of 50 (16%) vs 51 of 150 (34%) (p = < 0.05). The prevalence of FGR associated with the highest quartile of hPL (> 1.73 micrograms/ml) was 12.2% compared to 35% in the lower three quartiles (p = 0.025) and the prevalence of FGR associated with the highest quartile of SP1 (> 43 ng/ml) was 14% compared to 34.7% in the lower three quartiles (p = 0.018). Only one out of 21 infants (4.5%) whose mothers had each value in the highest quartile of hPL, estradiol, and SP1 was diagnosed with FGR compared to 58 out of 178 (32.6%) of the remaining infants (p = 0.007). CONCLUSIONS: In pregnancies of women at high risk for FGR, higher levels of estradiol, hPL, and SP1 at 18 weeks are associated with a decreased prevalence of FGR. This finding indicates that high levels of these hormones are related to a lower risk of FGR, but that low levels do not predict FGR. PMID- 9219459 TI - Cell division in placentas of appropriate and small-for-gestational-age infants. A flow cytometry study. AB - BACKGROUND: The purpose of this study was to examine if placentas of small- for gestational-age (SGA) and non-SGA infants differ with respect to proliferative cell activity. METHOD: Cell cycle distribution was studied in placentas from 181 SGA (birthweight < 10th percentile) and 528 non-SGA births by flow cytometry measurements of relative DNA content. RESULTS: The fraction of cells in various cell cycle phases (G1-, S- and G2-phases) did not differ with gestational age from 30 to 43 weeks in either of the groups. The placentas of the SGA infants had a significantly lower mean (+/-1 SEM) growth fraction than placentas of non-SGA infants (S-phase 5.2 +/- 0.2 vs 5.5 +/- 0.1, p = 0.05, and G2-fraction 5.4 +/- 0.2 vs 6.3 +/- 0.1, p < 0.001), but the overlaps of the distributions were large. Thus sensitivity, specificity and predictive values of low fractions did not differ substantially-from a purely random prediction of SGA. CONCLUSIONS: Cell division in the placenta is maintained until and beyond term. Placentas of SGA infants have on average, lower proliferative activity than placentas of non-SGA infants, but the difference is too small to be of predictive value in identifying intrauterine growth retardation. PMID- 9219460 TI - Serum concentrations of zinc, folate, vitamins A and E, and proteins, and their relationships to pregnancy outcome. AB - OBJECTIVE: To review the relationships between various laboratory measures relating to nutrition and pregnancy outcome. The data were obtained during the investigation entitled "Successive small-for-gestational-age births study". METHODS: A total of 289 pregnant women of the 1545 who participated in the study between 1986 and 1988 in Birmingham, Alabama, USA. The following determinations were done using the serum samples obtained at 18 and 30 weeks of gestation: zinc, folate, vitamins A and E, and proteins (alpha-2-macroglobulin, retinol-binding protein, prealbumin, and albumin). These laboratory values were correlated with various measures of pregnancy outcome including the incidence of fetal-growth retardation and maternal infections during the perinatal period and birth weight and Apgar score of infants. RESULTS: Serum folate concentrations showed positive relationships with the incidence of fetal-growth retardation as well as birth weight of infants, and alpha-2-macroglobulin was negatively correlated with birth weight. These relationships were significant after adjusting for factors previously known to affect the birth weight of infants. The concentrations of serum zinc, vitamins A and E, and proteins did not show significant correlation with measures of pregnancy outcome. CONCLUSION: Among the laboratory measures evaluated in this study, serum folate and alpha-2-macroglobulin concentrations correlated with pregnancy outcome. Further research is warranted to investigate the mechanism(s) of the relationship between serum alpha-2-macroglobulin and birth weight of infants. PMID- 9219461 TI - The relationship between maternal dietary intake and infant birthweight. AB - BACKGROUND: Zinc and folate are important for fetal growth. However, the relationship between the dietary intake of these nutrients and pregnancy outcome is not settled. METHODS: A prospective study was conducted to ascertain the relationship between maternal dietary zinc and folate intake (n = 1398), serum zinc and folate levels (n = 289), and infant birthweight. Twenty-four hour recalls were used to measure energy, zinc, folate and other nutrient intakes at 18 and 30 weeks of gestation. Subjects in the study were offered daily folic acid (1.0 mg) and iron (60 mg as ferrous sulfate) at enrollment. RESULTS: Maternal zinc nutriture as assessed by serum and dietary intake was not associated with birthweight or length of gestation. There was a small but significant positive association between maternal folate intake and adjusted infant birthweight (beta = 0.05, p = 0.03). The indirect measures of maternal nutritional status including maternal pre-pregnancy weight (beta = 8.0, p = 0.0001) and weight gain during pregnancy (beta = 18.1, p = 0.0001) were stronger predictors of adjusted infant birthweight as compared to energy intake and intake of zinc and folate. An increase of 320, 290, and 48 g in infant birthweight was associated with the 90th 10th percentile difference for pre-pregnancy weight, weight gain during pregnancy, and folate intake respectively. CONCLUSION: These results indicate that pre-pregnancy weight and weight gain during pregnancy are both strong predictors of infant birthweight. Folate intake, although significantly associated with birthweight, was a weak predictor while maternal intake of zinc and other nutrients was not associated with birthweight. PMID- 9219462 TI - Body proportions and early neonatal morbidity in small-for-gestational-age infants of successive births. AB - BACKGROUND: We wanted to examine if infants who were small for gestational age (SGA) at term had increased perinatal mortality or morbidity compared to non-SGA infants, and if this could be related to the infant's body proportions, or to whether the mother previously had delivered a low-birthweight infant ("repeater") or not ("non-repeater"). METHODS: From a cohort of 5722 para 1 and para 2 women, we compared perinatal mortality in 541 SGA (birthweight < 10th percentile) and 4737 non-SGA infants. From the same cohort, early neonatal morbidity was studied in 368 SGA and 462 control infants without congenital malformations. RESULTS: SGA infants had a 6.4 (95% CI: 2.6-15.7) higher risk of perinatal death than controls, but when infants who died with congenital malformations were excluded, this risk was not significantly increased. SGA infants were more often transferred to an intensive care unit than controls (1.7, 95% CI: 1.0-2.9). Among SGA births, infants with asymmetric body proportions (i.e. low ponderal index) more often had symptoms in the neonatal period (RR: 2.5; 95% CI: 1.4-4.3) and were more often transferred to an intensive care unit (3.4; 95% CI: 1.6-7.4) than symmetric SGA infants, whereas there were no differences between SGA infants of repeaters and non-repeaters. CONCLUSIONS: We found that SGA infants had higher perinatal mortality than controls, but this was due to a higher prevalence of congenital malformations. Among SGA infants without malformations, our results indicated increased neonatal morbidity in infants with asymmetric body proportions. PMID- 9219463 TI - Home environment and cognitive abilities in infants born small-for-gestational age. AB - BACKGROUND: In the present study we analyzed the relationship between home environment and cognitive abilities in small-for-gestational-age infants. METHOD: A group of 142 small-for-gestational-age infants and a control group of 172 appropriate-for-gestational-age infants were tested on the Fagan Test of Infant Intelligence at 7 months. The Home Screening Questionnaire was completed by the mothers when their infants were 13 months. RESULTS: The group of small-for gestational age infants had significantly lower scores on both the Fagan test (p < 0.05) and on the Home Screening Questionnaire (p < 0.01). A significant relation between the Fagan test score and the home score was found for the small for-gestational-age group (p < 0.05). When the home score was controlled for, the difference in mean Fagan score between the two groups of infants disappeared. CONCLUSIONS: It is suggested that small-for-gestational-age infants may be more vulnerable to adverse social conditions that infants born with a normal birthweight for gestational age. Results also suggest that cognitive impairments among small-for-gestational-age infants may be an effect of their social environments and their parents' general intelligence. Possible physical and neurological effects of intrauterine growth retardation may be less important for cognitive functioning. PMID- 9219464 TI - Growth and development during the first year in a cohort of low income term-born American children. AB - BACKGROUND: Infants born small for gestational age (SGA) are at risk for poor postnatal growth and development. This study evaluates biologic and environmental determinants of outcome during the first year of life in a cohort of low income term-born American infants. METHODS: Seven hundred and seventeen of 949 (76%) singleton births to women followed from early pregnancy were studied over their first year of life and measures of growth, home environment, physical and cognitive development were obtained. Infants were categorized as SGA or non-SGA based on birthweight < 15th percentile for gestational age. SGA and non-SGA children's outcomes were analyzed by race, gender and symmetry. RESULTS: SGA infants were demographically similar to non-SGA infants but significantly lower in mean maternal height, weight and education. Birthweight, crownheel length and head circumference were all significantly smaller in SGA infants. By age 1 year, the SGA children were still shorter, lighter and had smaller head circumferences than the non-SGA children though their rate of growth during the first year was significantly greater for length and head circumference. Cognitive functioning as measured by the Bayley Scales of Infant Development and the Fagan Test of Infant Intelligence did not differ significantly except for a lower Bayley Psychomotor Development Index (PDI) in SGA infants. Since most of these children live in economically disadvantaged households, any negative consequences of poor intrauterine growth may be influenced by postnatal environment and longer term follow-up will be necessary to assess this relationship. PMID- 9219465 TI - Small-for-gestational-age (SGA) infants born at term: growth and development during the first year of life. AB - BACKGROUND: The purpose was to compare growth patterns and psychomotor development of healthy small-for-gestational-age (SGA) and non-SGA infants, and identify factors predictive of outcome at 13 months of age. METHOD: A total of 265 SGA infants and 329 non-SGA controls were identified from a multicenter cohort of 5722 para 1 and 2 women who had been followed during pregnancy. The infants were examined at 2 days and at 13 months of age. Psychomotor development at 13 months was assessed with The Bayley Scale of Infant Development. RESULTS: The SGA infants showed partial catch-up growth, but had still lower (mean +/- SEM, p < 0.0001) weight (9750 +/- 65 vs 10505 +/- 67 g), crown-heel length (75.9 +/- 0.2 vs 77.5 +/- 0.2 cm) and head circumference (46.9 +/- 0.1 vs 47.7 +/- 0.1 cm) than the non-SGA infants at 13 months. The SGA children scored equally well on the motor (PDI 106.8 +/- 1.0 vs 107.2 +/- 0.8) but lower on the mental scale (MDI 112.1 +/- 0.8 vs 116.5 +/- 0.7, p < 0.0001) of the Bayley Scale, and the asymmetric SGA scored lower than the symmetric SGA infants (MDI 110.2 +/- 1.3 vs 113.3 +/- 0.9, p = 0.05). In a multivariate regression analysis the parents' growth parameters had the greatest effect on growth measures at 13 months while education and maternal smoking had no significant effect. SGA vs non-SGA status had the greatest effect on growth velocities during infancy. For mental development only SGA vs non-SGA status and the mothers' education made significant contributions, but only accounted for 6% of the variance. CONCLUSION: The negative impact of intrauterine factors on growth are partly abolished by catch-up growth during infancy, and growth parameters at one year of age are mostly determined by genetic factors even in SGA infants. Decreased intrauterine growth may possibly have a negative effect on brain growth and mental developmental potential. PMID- 9219466 TI - Prospects of the collaborative small-for-gestational-age birth study for the five year old follow-up study. PMID- 9219467 TI - Coronary artery bypass conduits: review of current status. AB - Coronary artery bypass grafting (CABG) is the most common procedure performed in adult cardiovascular surgery today. In our Department of Surgery at Baylor College of Medicine, we have experienced, as have most large programs, a trend to older patients, more comorbidity, worse ventricular function, and more redo CABG procedures. Along with this shift has come an evolution in surgical techniques, cardioplegia and choice of coronary bypass graft conduits. The benefit of using an internal mammary artery (IMA) to the left anterior descending coronary artery (LAD) appears irrefutable at this time. Data for multiple arterial grafts is still evolving and conduit choice for other than the IMA to LAD graft is often debated. The purpose of this article is to review the current literature on conduit choice and to allow a rational, data driven approach to graft choice. PMID- 9219468 TI - Clinical importance of measuring coronary graft flows in the revascularized heart. Ultrasonic or electromagnetic? AB - BACKGROUND: In the past, routine coronary graft flow measurement at the end of coronary artery bypass grafting (CABG) was not universally adopted by cardiac surgeons due to the lack of reliable flow measurement techniques. The purpose of this study was to investigate the efficacy of ultrasonic and electromagnetic techniques in coronary graft flow measurements and to determine the relationship, if any, between intraoperative ultrasonic or electromagnetic coronary graft flows and postoperative early clinical outcome. METHODS: We studied 66 consecutive patients who underwent elective CABG using internal thoracic artery (ITA) and reversed saphenous vein graft (SVG) conduits. All patients were males with the mean age of 65 +/- 1 yrs (range = 45 to 80 yrs). Coronary bypass graft flows (both ITA and SVG) were determined by the use of both ultrasonic and electromagnetic flowmeters. In addition, the flow waveform pattern was continuously recorded and analyzed with the ultrasonic technique. In this prospective non-randomized study, the following variables were considered in the forward stepwise multivariate regression analysis of the data: age, weight, body surface area, ejection fraction, perfusion and ischemia times, number of grafts, amount of allogenic banked blood, platelets, fresh frozen plasma transfusions, cardiac output/index, ultrasonic (USF) and electromagnetic flows (EMF), length of intensive care unit (ICU) and hospital stays, and early (30-day) mortality. RESULTS: Based on their location, 226 grafts were divided into four groups: (I) ITA to left anterior descending (LAD) (n = 66) 34 +/- 2.5 ml/min USF and 45 +/- 4.4 ml/min EMF; (II) SVG to circumflex (CX) (n = 62) 33 +/- 2.4 m/min USF and 58 +/- 4.9 ml/min EMF; (III) SVG to diagonal (DIAG) (n = 37); 30 +/- 3.5 ml/min USF and 50 +/- 6.0 ml/min EMF; (IV) SVG to right coronary artery (RCA) (n = 61); 36 +/- 3.1 ml/min USF and 56 +/- 5.3 ml/min EMF. Electromagnetic flow measurements were higher than USF values in all locations (p < 0.05). Difficulties in obtaining proper contact with the vessel wall and finding suitable size probes were major drawbacks in measurement of ITA graft flow by the use of electromagnetic technique. All flow measurements were done within 10 minutes or less. There was no demonstrable correlation between the length of stay (ICU and hospital), and coronary graft flows at the ITA to LAD, SVG to DIAG, or SVG to CX locations. However, ultrasonic coronary graft flows at the SVG to RCA location had a significant inverse correlation with the length of ICU and hospital stays (r = -0.45, p < 0.0005 for both). Early mortality was unaffected by the intraoperative coronary graft flow values (p = NS). CONCLUSIONS: The ultrasonic flowmeter is well-suited for intraoperative assessment of arterial and venous coronary graft flows at the completion of CABG. There is a real potential for using intraoperative graft flow values to predict early outcome after coronary bypass. PMID- 9219469 TI - Surgical treatment of coronary artery aneurysm after percutaneous transluminal coronary angioplasty (PTCA). AB - The case of a 65-year-old man who developed a coronary artery aneurysm at the site of percutaneous transluminal coronary angioplasty (PTCA) in the proximal right coronary artery (RCA) is described. The coronary artery aneurysm was resected and a coronary artery bypass to the distal RCA was grafted to avert thromboembolism. PMID- 9219470 TI - Position-related factors in mitral and tricuspid bioprostheses degenerative changes. AB - We report clinicopathological findings in 15 patients in whom the same bioprosthesis (BP) had been implanted simultaneously in both mitral and tricuspid positions. The aim of the study was to investigate whether position-related factors played an important role in BP degeneration. There were 14 women and 1 man with a mean age of 34 +/- 11 years. The indications for the initial operation were rheumatic in 14 cases and endocarditis in one patient. The mean interval before reoperation was 7.5 +/- 3.3 years. Predominant cause of reoperation was: structural deterioration of both mitral and tricuspid BPs (6), mitral regurgitation (5), tricuspid BP dysfunction (1), para-aortic leak (1), mitro aortic thrombi (1). Calcific deposits were the principal cause of early deterioration of mitral BPs and the major cause of late tricuspid BPs dysfunction. This lesion was predominantly related to local factors. Cuspal tears were the principal cause of late (> 9 yrs) mitral BP failure and most probably related to mechanical stress. Extensive fibrosis affected only tricuspid bioprostheses. In 7 patients more extensive degenerative changes occurred in bioprostheses in the mitral rather than the tricuspid position (Group I). However, in the remaining eight the magnitude of the changes was very similar in the two positions (Group II). The interval before reoperation was significantly longer in patients of Group II (9.8 yrs, range 5-13) than patients in Group I (4.9 yrs, range 3-6), (p < 0.01). We concluded that position-related factors exert a major role in bioprosthetic failure. These factors are more deleterious in the mitral position than in the tricuspid position. PMID- 9219471 TI - Pulsatile and nonpulsatile extracorporeal circulation using Capiox E terumo oxygenator: a comparison study with Ultrox and Maxima membrane oxygenators. AB - An open randomised, prospective study was undertaken on 90 patients who underwent routine myocardial revascularization. The aim of the study was to demonstrate that the Capiox E polypropylene fiber membrane oxygenator with a conventional single pulsatile/nonpulsatile blood pump for cardiopulmonary bypass (CPB) was comparable in performance to that of the Maxima and the Ultrox membrane oxygenators using a double pump system. The patients were divided into six groups according to perfusion mode and oxygenator type. Laboratory parameters, fluid balance and oxygenation was examined at set times before during and after cardiopulmonary bypass. Net fluid input was lower in the Capiox E groups regardless of perfusion mode: 2932 +/- 562 ml (Capiox E), compared to 3646 +/- 531 ml (Ultrox) and 3593 +/- 582 ml (Maxima). Net fluid balance 1288 +/- 534 ml was lowest in the Capiox/NP group, compared to 1604 +/- 460 ml (Ultrox/NP) and 1881 +/- 594 ml (Maxima/NP), (p < 0.05). The higher net fluid balance in the Capiox E/PP group 1649 +/- 580 ml compared to 1592 +/- 583 ml (Ultrox E/PP) and 1494 +/- 542 ml (Maxima/PP) was attributed to a technicality whereby the recommended priming volume of the Capiox E oxygenator was exceeded for safety reasons. The values of plasma free Hb were slightly higher in the PP than NP groups: Maxima/PP 80 mg/dl, /NP 50 mg/dl; Ultrox/PP 62 mg/dl, /NP 48 mg/dl; Capiox E/PP 55 mg/dl, /NP 48 mg/dl. The FiO2 was higher in the Capiox E groups 0.77 (PP) and 0.88 (NP) compared to Maxima/PP (0.66), /NP (0.65) and Ultrox/PP (0.64), /NP (0.63). Reciprocally, the venous saturation was higher in the Ultrox and Maxima groups compared to Capiox E at end of CPB. The study demonstrated that the CapioxE oxygenator with a single blood pump system can compare to the Maxima and Ultrox oxygenators with a double blood pump for CPB with regard to blood handling, oxygenation and fluid balance in routine cardiac surgery. PMID- 9219472 TI - Individualized repair of the left atrioventricular valve in spectrum of atrioventricular septal defect. AB - From September 1977 to October 1995, 287 patients with atrioventricular septal defect (AVSD) aged from 2 months of 21 years underwent total repair in Kardiocentrum in prague. In 97 patients complete, in 20 transitional and in 170 patients partial form of AVSD was present. The repair consisted of closure of the defect and individually modified reconstruction of two atrioventricular (AV) orifices. In cases with a common orifice a two-patch technique was used. Fixation of undivided anterior and posterior common leaflets to patches in an appropriate level was essential in combination with complete closure of the cleft. Incomplete closure of the cleft was performed if potentially stenotic morphology was present. Commissuroplasty with pladgeted mattress stitches was done in patients with dilated annulus and commissuroplasty with a single stitch was performed if the annulus was not dilated. The methods were similar in cases with two AV orifices. The AV valve repair was difficult in the presence of severe regurgitation in valves with potentially stenotic morphology. Of the 287 operated patients 26 (9.1%) died during the early postoperative period. Mortality was 19.6% in the complete form and 3.7% in the partial and transitional forms. The mortality depended on morphology of the left atrioventricular valve. Potentially stenotic valvar morphology represented an important risk factor for death and reoperation. It was necessary to reoperate on 18 (6.3%) patients for significant "mitral" valve regurgitation. Reconstruction of a competent left AV valve is the most important step of AVSD repair which must always be modified according to individual morphological and functional abnormalities. PMID- 9219474 TI - The significance of oncometry for infusion therapy during pediatric heart surgery. AB - The colloid osmotic pressure (COP) is not routinely assessed during pediatric heart surgery. Two cases of unrecognized hyperoncotic states associated with renal failure have been observed after pediatric heart surgery. We studied the hypothesis that the COP cannot be estimated from the total plasma protein (TPP) or albumin level. The course of COP and its correlation to the TPP and albumin level were investigated in 25 children undergoing elective heart surgery. Infusion therapy was performed solely on the basis of clinical parameters and TPP/albumin levels. COP values were determined in a blinded fashion at the end of the study. No correlation between TPP/albumin and the COP could be determined preoperatively. On arrival at the ICU correlation was strong. A weak correlation was observed at 24 hours and 48 hours after surgery. However, the observed wide range of the confidential bands indicates that the COP cannot be estimated correctly, neither from the TPP, nor from the albumin level. Due to colloidal oversubstitution COP was significantly increased compared to preoperative level at 48 hrs following surgery. As estimation of COP from TPP or albumin level is inaccurate, oncometry should be performed during pediatric heart surgery. PMID- 9219473 TI - Long-term results of heart valve replacement with bileaflet prostheses. AB - Bileaflet cardiac prostheses (St. Jude, CarboMedics, Duromedics, Bicarbon, Jyros) have shown a low incidence of complications and good haemodynamic performance. In the last twelve years, 783 bileaflet prostheses were implanted in 690 patients at our Institution. The population of our study comprises 591 bileaflet prostheses (418 CarboMedics, 124 St. Jude, 49 Bicarbon) implanted in the mitral (MVR) (n = 305) or aortic (AVR), (n = 286) position. The follow-up study evaluated 292 male and 295 female patients with age ranging from 13 and 79 years (mean 50.4 +/- 14.7 years). Hospital mortality was 6.6%. Follow-up was 97% complete, with 1822 +/- 33 patient/years and a mean follow-up of 37 months (range 1 to 144 months). Twelve years actuarial freedom from complication according to prosthetic site were calculated as follows (linearized rates in parentheses): late mortality AVR 97.6% +/- 0.6% (2.3%), MVR 96% +/- 0.5% (2.1%); thrombosis AVR 100%, MVR 96% +/- 0.9% (0.8%); embolism AVR 97% +/- 0.5% (1.5%), MVR 96.6% +/- 0.7% (1.8%). Global freedom from anticoagulant-related haemorrhage was 95% +/- 1.2% (2.3%) and 94.5% +/- 0.7% (2.2%) following AVR, 94 +/- 0.6% (2.1%) following MVR. The difference of the haemorrhagic risk for prosthetic site was not significant (p > 0.05). Functional improvement was confirmed by the low postoperative NYHA functional class. According to our results, cumulative experience with bileaflet valves has shown very good long-term results in term of low rate of complication, long-term survival and quality of life. PMID- 9219475 TI - A case of Turner's syndrome associated with partial anomalous pulmonary venous return complicated by dissecting aortic aneurysm and aortic regurgitation. AB - We report a successful surgical case with Turner's syndrome associated with partial anomalous pulmonary venous return (PAPVR) complicated by aortic dissection and aortic regurgitation without coarctation of the aorta. The patient, a 30-year-old woman, is of a short stature who was diagnosed with Turner's syndrome at the age of 12. She has suffered from dyspnea and edema of the legs since a year ago and was admitted to our hospital in June 1994 as echocardiography revealed rapid dilatation of ascending aorta and aortic regurgitation. A chest X-ray showed cardiothoracic ratio of 63% and transesophageal echocardiogram revealed that ascending aortic diameter was extended up to 60 mm at its maximum and that it was possible to distinguish true lumen from false lumen. The aortic arch was found to be normal. Also revealed by cardiac catheterization was drainage of the left upper pulmonary vein to the innominate vein. The L-R shunt ratio was 2.2. The surgery was performed by the Bentall method. The composite graft with a 21 mm St. Jude Medical prosthetic heart valve placed on the annulus of aortic valve. The ostiums of the coronary arteries were directly anastomosed to the composite graft with Carrel patch. After declamp of the aorta, the left pulmonary vein was directly anastomosed to the left atrial appendage without causing stenosis. The postoperative course was uneventful, and the cineangiogram after surgery demonstrated successful repair. Reports of cases of Turner's syndrome like this are sparse. PMID- 9219476 TI - Management of radiation-induced occlusive arterial disease: a reassessment. AB - BACKGROUND: The goal of this study was to evaluate the operative hazards, therapeutic procedures, and late results of arterial reconstruction for radiation induced occlusive disease. METHODS: Twenty-five patients were referred to our institution for radiation-induced occlusive arterial disease. Group 1: carotid artery stenosis or occlusion was encountered in seven patients. The nine procedures employed included percutaneous transluminal angioplasty (PTA) (n = 2), carotid endarterectomy (n = 3), vein or prosthetic bypass (n = 4). Group 2: four patients presenting with subclavian and axillary artery occlusion were treated with a common carotid to brachial artery vein bypass, one after unsuccessful PTA. Group 3: Thirteen patients had aorto-iliac occlusion. Initial management included medical treatment (n = 1), PTA (n = 2), aorto-bifemoral bypass (n = 4), aortofemoral and iliofemoral bypass (n = 1 each), axillofemoral bypass (n = 3), femorofemoral bypass (n = 1). Group 4: One patient had femoral artery occlusion treated with PTA. RESULTS: Group 1: One of two PTA was successful. Endarterectomy or bypass were successful in all cases. One late vein bypass stenosis was treated by venous patch angioplasty. Group 2: All vein bypasses were successful. Group 3: Limb salvage was achieved in all patients but eight required repeat operations for prosthetic sepsis (n = 3), restenosis (n = 3), or thrombosis (n = 12). Two patients died of late sepsis. Group 4: Outcome after PTA was successful. CONCLUSIONS: 1) Surgery for radiation-induced arterial lesions is difficult because of arterial, periarterial, and cutaneous sclerosis. Some patients, however, are amenable to PTA or endarterectomy. When bypass is necessary, anastomosis should be performed in healthy arteries, for instance, the thoracic aorta for the proximal anastomosis, or the brachial artery approached through a lateral mid-arm incision. 2) The risk of early or late graft infection is enhanced by the presence of tracheostomy, colostomy, or ureterostomy and by repeat operation for thrombosis. PTA, endarterectomy, or vein bypass should be preferred whenever feasible. When prosthetic material is unavoidable, prevention of infection should include the use of omentoplasty, remote bypass, antibiotic bonded grafts or, in the case of major sepsis, allografts. 3) As restenosis remains a frequent complication, annual clinical and Duplex-scan surveillance is mandatory. PMID- 9219477 TI - Apico-aortic shunt: a support technique during surgery on the descending thoracic aorta. AB - To find out whether apico-aortic shunt may become an alternative support technique during surgery on the descending thoracic aorta, performance between apico-aortic shunt and aorto-aortic shunt was compared. In 5 sheep weighing 20-25 kg, apico-aortic shunt and aorto-aortic shunt were instituted with covalently bonded heparin coated polyvinyl tube (internal diameter 5 mm). After clamping the descending thoracic aorta, apico-aortic shunt and aorto-aortic shunt were opened for 30 minutes each. Proximal pressure was elevated to 200 mmHg and distal pressure was fallen to 55 mmHg after clamping the descending thoracic aorta. Opening of apico-aortic shunt and aorto-aortic shunt decreased proximal pressure to 178 +/- 14.8 and 173 +/- 12.0 mmHg, respectively (p = 0.57), and raised distal pressure to 82.4 +/- 7.8 and 90.0 +/- 3.5 mmHg, respectively (p = 0.83). The baseline blood flow of the descending thoracic aorta was 1.4-1.5 l/min. Apico aortic shunt and aorto-aortic shunt were 0.76 +/- 0.16 and 0.80 +/- 0.22 l/min, respectively (p = 0.67). Blood gas tension, pH and BE measurement showed no significant change and difference between apico-aortic shunt and aorto-aortic shunt, and before and after clamping the descending thoracic aorta. Using apico aortic shunt, interposition of bioprosthetic valved conduit in the descending thoracic aorta in 10 sheep was successfully performed without paraplegia and any other complications. We concluded that apico-aortic shunt may become an alternative support technique during surgery on the descending thoracic aorta in some specific situations. PMID- 9219478 TI - Cells in pseudointimal hyperplasia is migrated from extravascular space. AB - BACKGROUND: The aim of this study was to investigate the origin of the pseudointima (PI) formed in polytetrafluoroethylene (PTFE) tube grafts after implantation into the inferior vena cava (IVC) of rabbits. METHODS: A segment of the IVC of rabbits was replaced by PTFE tube graft (3 cm long, 3 mm inner diameter, 30 microns internodal distance, 0.3 mm thickness). The experimental group was divided into two groups as follows: (Group A) non-wrapped, (Group B) wrapped the outer wall of PTFE with impermeable vinyl. RESULTS: Grafts were harvested at three weeks after implantation and subjected to the following studies: patency, ultrastructural studies by light microscopy (LM) and immunostaining, scanning and transmission electron microscopy (SEM & TEM). The grafts were patient but the lumen of the control group was narrowed by PI. LM and immunostaining studies revealed the presence of thick PI composed of spindle-type cells in Group A, bust almost no PI in Group B. Only few erythrocytes, macrophage and protein-fibrin matrix was found in Group B. Endothelial like cell coverage, judged by SEM, was observed in only Group A. Only some macrophages and platelets were shown in the graft surface in Group B. TEM of PI revealed the presence of VSMCs, myofibroblasts and outer surface of grafts revealed the presence of myofibroblast in Group A. CONCLUSIONS: The formation of PI suppressed by blocking the cellular migration from perigraft space suggest that PI was mainly originated by myofibroblast located in the perigraft space. PMID- 9219479 TI - Experience with the use of polytetrafluoroethylene pericardial membrane (preclude pericardial membrane) for retroperitoneal closure after abdominal aortic surgery. AB - OBJECTIVE: The purpose of the present publication is to assess the outcome after closure of the retroperitoneum with a patch of polytetrafluorethylene surgical membrane (Preclude Pericardial Membrane. PPM) after abdominal aortic surgery. EXPERIMENTAL DESIGN: Retrospective review of 7 operated patients with a patch of PPM covering their abdominal aortic synthetic graft. Twenty-five months of follow up. SETTING: Institutional and University Hospital. PATIENTS AND PARTICIPANTS: Between June 1993 and February 1995, in 7 consecutive male patients (mean age 62 years) a patch of PPM was applied to their retroperitoneum after undergoing surgery in their infrarenal abdominal aorta. One patient had total obliteration of the distal abdominal aorta (Leriche syndrome) and the remaining 6 had a small infrarenal abdominal aortic aneurysm measuring in transverse diameter between 4 and 5 centimetres. INTERVENTIONS: The patient with Leriche Syndrome had an end to end aorto bifemoral bypass graft and the 6 cases with abdominal aortic aneurysm underwent resection of the aneurysm plus interposition of a straight vascular graft (3 cases), straight vascular graft and extension with other graft to the femoral artery (1 case) and bifurcated aorto-bifemoral graft (2 cases). MEASURES: Clinical outcome and evolution of the patients, absence of complications derived from the PPM. RESULTS: There was no hospital mortality. All patients are alive after a mean follow-up of 25 months. Two patients were reoperated in the early postoperative period, one of them required a limited resection of the jejunum. There have been no complications related to the PPM. CONCLUSIONS: In order to avoid secondary aorto-intestinal fistulas, in cases where complete coverage and isolation of an artificial vascular graft in the abdominal aorta can not be achieved, the use of a sheet of PPM separating the arterial graft from the gastrointestinal system may be an useful alternative. PMID- 9219480 TI - Postoperative inflammatory reactions to sealed Dracon prostheses: a comparison of Gelseal and Hemashield. AB - OBJECTIVE: The purpose of this study was to assess and compare inflammatory reactions to two types of sealed vascular prostheses used in the repair of abdominal aortic aneurysm. EXPERIMENTAL DESIGN: Randomized prospective study with a follow-up period of 21 days. SETTING: University hospital study. PATIENTS: Sixty patients admitted for elective abdominal aortic aneurysm repair. INTERVENTION: Each patient underwent repair of abdominal aortic aneurysm with either a gelatin-sealed knitted Dacron prosthesis (Gelseal, Vascutek, n = 30) or a collagen-sealed woven Dacron prosthesis (Hemashield, Meadox Medicals, n = 30). MEASURES: As indicators of the presence of inflammation, the peripheral blood cell count, platelet count, and plasma C-reactive protein (CRP) concentration were determined in each patient on postoperative days (PODs) 1, 7, 14 and 21. Body temperature was measured every 8 hours. RESULTS: Patients in both groups revealed a low-grade fever and leukocytosis during the first 7 PODs. Significantly higher leukocyte counts were demonstrated in the Gelseal group on PODs 7 and 14. However, no difference in the leukocyte count, CRP concentration, or body temperature existed between the two groups on POD 21. Biphasis changes in the platelet count were observed in both groups. No complications were encountered. CONCLUSION: The Gelseal prosthesis was associated with higher inflammatory response during the second week following implantation, but this difference had resolved by POD 21. This finding may reflect an inflammatory reaction against impregnated gelatin. PMID- 9219481 TI - Blood flow velocity measurements in the optic nerve of glaucomatous patients by elaboration of a constant to evaluate the evolutive risk. AB - METHODS: We performed Echo color Doppler examinations on 100 glaucomatous patients (197 eyes). Ophthalmic artery and central retinal artery were particularly considered. The study included eyes in every level and type of glaucoma, and in various types of treatment (topical therapy, Argon laser trabeculoplasty, Argon/YAG/surgical iridectomy, trabeculectomy). The parameters we considered were: ocular tension, anterior chamber angle, optic disc excavation, visual field, refraction and vascular risk factors. We measured the flow values of the ophthalmic artery and of the central retinal artery, i.e. maximal systolic velocity, diastolic velocity and vascular resistance index. RESULTS: We observed that if we divide the value of ocular tension by the systolic velocity and multiply the result by the resistance index, we obtain a number which may be considered as a prognostic factor of evolutive risk. We compared our cases with a population free from glaucomatous and cardiovascular problems. PMID- 9219482 TI - Exclusion of an internal carotid aneurysm by a covered stent. AB - The authors report a case of a patient with thromboembolic strokes caused by a high internal carotid artery aneurysm. Considering the position and the anatomic structure of this aneurysm the sac was excluded transluminally by placing an endovascular covered stent. PMID- 9219483 TI - Aneurysm of the right colic artery. AB - Aneurysms of the superior mesenteric artery branches are rare. Spontaneous intra abdominal hemorrhage resulting from rupture of a visceral artery aneurysm is difficult to diagnose and carries high mortality. We present a case in which a rupture of the right colic artery aneurysm presented as an acute abdomen. Diagnosis was established intraoperatively. Angiography and a high degree of suspicion are valuable diagnostic tools. Exact etiology was not determined in our case. Because of high risk of rupture, aneurysms of the superior mesenteric artery branches should be resected. PMID- 9219484 TI - Horton giant cell arteritis of the legs. Report of a case. AB - Horton giant cell arteritis of the legs is a very rare and unusual occurrence. A very interesting case of acute ischemia of the right leg in a 51-year-old woman treated with emergency thromboendarterectomy is described. Histological findings led to the diagnosis of Horton giant cell arteritis and the patient was submitted to steroid and vascular therapy. Good results were obtained and follow-up after five years confirms the good general condition of the patient. PMID- 9219486 TI - Surgical therapy of malignant thymoma. AB - OBJECTIVE: To give a modern concept for treating tumours of the thymic gland. EXPERIMENTAL DESIGN: Retrospective analysis of all patients treated for thymic disease in the last 6 years with a mean follow-up of 22.3 months (range: 3-71 months). SETTING: All patients were admitted to the Department of Surgery of the University of Cologne with an average hospitalisation period of 14 days. Patients with myasthenia gravis were transferred to the intensive care unit postoperatively. PATIENTS: A total of 34 patients were treated by radical thymectomy. All patients had a routine chest roentgenogram and a computed tomography. In 13 patients a malignant tumour of the thymic gland was diagnosed. In 6 patients the main symptom was myasthenia gravis. INTERVENTIONS: All tumours were successfully resected by an upper median sternotomy which was extended through a Kocher incision (n = 2) or a complete sternotomy (n = 3). In one patient the tumour was unresectable. RESULTS: In four patients histological examination showed a thymic carcinoma and in 9 patients an invasive thymoma. Four patients died during follow-up period. Stage II to IV thymoma should receive postoperative radiotherapy and an additionally chemotherapy in stage IV. Cumulative survival rate for a five years follow-up independent of the tumour stage was 46%. CONCLUSIONS: Malignant tumours of the thymic gland are a rare entity and should be treated by radical resection of the tumour and the adjacent tissue. In stage II and III postoperative radiation can lower the risk of recurrence. In stage IV radiation should combined with chemotherapy. Long-term follow-up is mandatory. PMID- 9219485 TI - Use of 111In-DTPA-octreotide scintigraphy in the diagnosis of neuroendocrine and non-neuroendocrine tumors of the lung. Preliminary results. AB - BACKGROUND: The authors report their preliminary experience and results of the use of 111In-DTPA-octreotide scintigraphy (octreoscan) in the staging of neuroendocrine and non-neuroendocrine tumors of the lung. MATERIALS AND METHODS: From July 1995 to May 1996 twenty-six scintigraphic studies were performed in patients affected by lung cancer at the Department of Thoracic Surgery and at the Service of Nuclear Medicine of the University of Turin. RESULTS: Scintigraphy made it possible to detect the lesion in all the patients affected by neuroendocrine tumors and in 63.2% of the patients affected by non-neuroendocrine neoplasm of the lung. Scintigraphy also revealed mediastinal lymphnodal metastases in patients in which thoracic CT scan was negative: this result was confirmed by postoperative TNM. CONCLUSIONS: The authors stress the importance of 111In-DTPA-octreotide scintigraphy in a correct procedure of staging of neuroendocrine and non-neuroendocrine tumors of the lung and in the follow-up of neoplastic patients. PMID- 9219487 TI - Serotonin reuptake inhibitor discontinuation syndrome: a hypothetical definition. Discontinuation Consensus panel. AB - Adverse events following discontinuation from serotonin reuptake inhibitors (SRIs) are being reported in the literature with increasing frequency; the frequency and severity of these symptoms appear to vary according to the half life of the SRI, e.g., the incidence appears higher with the shorter half-life agents than with fluoxetine, which has an extended half-life. Yet, there have been no systematic studies of the phenomenon to date. Therefore, a group of experts convened in Phoenix, Arizona, to develop a clear description or definition of the phenomenon based on these reports. The SRI discontinuation syndrome, referred to as "withdrawal symptoms" in many anecdotal case reports, is distinctly different from the classic withdrawal syndrome associated with alcohol and barbiturates. Anti-depressants are not associated with dependence or drug seeking behavior. SRI discontinuation symptoms tend to be short-lived and self limiting, but can be troublesome. They may emerge when an SRI is abruptly discontinued, when doses are missed, and less frequently, during dosage reduction. In addition, the symptoms are not attributable to any other cause and can be reversed when the original agent is reinstituted, or one that is pharmacologically similar is substituted. SRI discontinuation symptoms, in most cases, may be minimized by slowly tapering antidepressant therapy, but there have been several case reports where symptoms occurred consistently even through repeated attempts to taper therapy. Physical symptoms include problems with balance, gastrointestinal and flu-like symptoms, and sensory and sleep disturbances. Psychological symptoms include anxiety and/or agitation, crying spells, and irritability. Further analyses of data bases and clinical studies are needed to define this proposed syndrome more clearly. PMID- 9219489 TI - Newer antidepressants and the discontinuation syndrome. AB - Data on discontinuation phenomena associated with serotonin selective reuptake inhibitors (SSRIs) are derived primarily from (1) published case reports, (2) data bases of adverse drug reactions that have been spontaneously reported to national monitoring bureaus, and (3) clinical studies of drug discontinuation. Some of the symptoms seen on SSRI discontinuation, such as nausea, lethargy, insomnia, and headache, are similar to those reported with tricyclic discontinuation. However, SSRI discontinuation is also associated with novel symptom clusters, including problems with balance, sensory abnormalities, and possibly aggressive and impulsive behavior. Although generally mild and short lived, discontinuation symptoms can be severe and chronic and have a major impact on the patient's lifestyle. The incidence of discontinuation symptoms varies widely among the different SSRIs; the highest rate is seen with paroxetine. The variation in incidence might be explained by the different pharmacokinetic and pharmacodynamic profiles of the SSRIs. PMID- 9219488 TI - Antidepressant discontinuation: a review of the literature. AB - Sudden or tapered withdrawal from treatment with antidepressants, including monoamine oxidase inhibitors (MAOIs), tricyclic antidepressants (TCAs), and serotonin selective reuptake inhibitors (SSRIs), can produce phenomena consisting of somatic and psychological symptoms. The literature about these discontinuation phenomena consists mainly of case reports and a limited number of controlled prospective studies. The symptoms are generally mild and transient for the TCAs and the SSRIs but may be serious for the MAOIs. They are much more common with a shorter acting SSRI, such as paroxetine, than with the longer acting agent fluoxetine. Because the symptoms of antidepressant discontinuation include changes in mood, affect, appetite, and sleep, they are sometimes mistaken for signs of a relapse into depression. Thus, it is important to directly question patients about new symptoms that occur during antidepressant discontinuation to optimally manage treatment discontinuation. PMID- 9219490 TI - Possible biological mechanisms of the serotonin reuptake inhibitor discontinuation syndrome. Discontinuation Consensus Panel. AB - Although the number of documented serotonin reuptake inhibitor (SRI) discontinuation reactions is increasing, to date no systematic studies have been completed; therefore the mechanism of action for these reactions is not clearly understood. However, several hypotheses have been proposed. Researchers have postulated that discontinuation events result from a sudden decrease in the availability of synaptic serotonin in the face of down-regulated serotonin receptors. In addition, other neurotransmitters, such as dopamine, norepinephrine, or gamma-aminobutyric acid (GABA), may also be involved, although little research in this area has been published. Individual patient sensitivity, i.e., genetics or cognitive mindset, may also be a factor in SRI discontinuation phenomena. Finally, experts have hypothesized that since some symptoms associated with paroxetine withdrawal are similar to those of tricyclic antidepressant discontinuation, they may be caused by cholinergic rebound. PMID- 9219491 TI - Physicians' knowledge of antidepressant withdrawal effects: a survey. AB - BACKGROUND: While the incidence of discontinuation events in controlled studies of serotonin reuptake inhibitors ranges between 34.5% and 86%, only a small number of discontinuation reactions are reported to national data bases of spontaneously reported adverse drug reactions. It was hypothesized that the disparity was due to lack of knowledge amongst physicians about the potential for antidepressant discontinuation reactions. METHOD: Therefore, a questionnaire was mailed to 100 psychiatrists and 100 general practitioners (GPs) in northeast England to assess the knowledge base and to validate this assumption. RESULTS: Fifty psychiatrists (50%) and 53 GPs (53%) responded to the questionnaire. Of the respondents, 36 (72%) of the psychiatrists and 16 (30%) of the GPs were aware that patients may experience antidepressant discontinuation events; 33 (66%) psychiatrists and 22 (42%) GPs had had experience with patients who had discontinuation symptoms; and 10 (20%) psychiatrists and 9 (17%) GPs said they always caution patients about the possibility of discontinuations events. CONCLUSION: According to the results of the survey, a sizable minority of physicians denied being confidently aware of the existence of antidepressant withdrawal symptoms. Education about discontinuation reactions, including the hallmark features, symptoms, and course, is needed for both psychiatrists and family practice physicians. PMID- 9219492 TI - Antidepressant noncompliance as a factor in the discontinuation syndrome. AB - Compliance is generally defined as the extent to which a patient adheres to a treatment regimen and, specifically, takes medication as prescribed. While little research is available about the number of patients who consistently skip antidepressant doses, the literature indicates that about 30% of patients discontinue treatment suddenly within the first month. Both missed doses and abrupt stoppage of treatment place a patient at risk for experiencing discontinuation symptoms. A variety of reasons ranging from forgetfulness to lack of knowledge about the importance of taking every dose may lead to nonadherence to an antidepressant regimen. By spending time on patient education, providing reasons why patients should take every antidepressant dose, discussing alternative treatments, and conveying empathy, support for, and understanding of the patient, physicians may be able to minimize noncompliance and consequently decrease the likelihood that a patient may experience discontinuation symptoms. Discontinuation of an antidepressant can cause a patient to be irritable, experience severe dizziness, or act emotionally absent, which may have a sustained adverse impact both on job performance and on family and social relationships. PMID- 9219493 TI - Clinical management of antidepressant discontinuation. AB - To minimize the symptoms of antidepressant discontinuation, gradual tapering is necessary for all serotonin reuptake inhibitors (SRIs) except fluoxetine, which has an extended half-life. Agents with shorter half-lives such as venlafaxine, fluvoxamine, and paroxetine should be tapered gradually. Discontinuation symptoms, which frequently emerge after abrupt discontinuation or intermittent non-compliance and, less frequently, during dose reduction, are generally mild, short-lived, and self-limiting but can be distressing and may lead to missed work days and decreased productivity. The symptoms may be somatic (e.g., dizziness and light-headedness; nausea and vomiting; fatigue, lethargy, myalgia, chills, and other flu-like symptoms; sensory and sleep disturbances) or psychological (anxiety and/or agitation, crying spells, irritability). Mild symptoms can often be treated by simply reassuring the patient that they are usually transient, but for more severe symptoms, it may be necessary to reinstitute the dosage of the original antidepressant and slow the rate of taper. Symptoms of discontinuation may be mistaken for physical illness or relapse into depression; misdiagnosing the symptoms may lead to unnecessary, costly tests and treatment. Thus, health care professionals need to be educated about the potential adverse effects of SRI discontinuation. PMID- 9219494 TI - Defining primary care. Empirical analysis of the National Ambulatory Medical Care Survey. AB - OBJECTIVES: Efforts to contain health care costs have increased interest in defining which specialties provide primary care and in developing tools to assess the delivery of primary care services. METHODS: Using data from the 1985-1991 National Ambulatory Medical Care Surveys, the authors examined the activities of 29 physician specialty groups to determine whether the recent Institute of Medicine definition of primary care could be operationalized. Ten elements were identified that addressed comprehensiveness (first-contact care, a Herfindahl Index, previous contact for other problems, prevention, and care through the life cycle), coordination (referrals), continuity (any previous contact), and accessibility (care provided to black patients, those on Medicaid, and patients in rural areas). RESULTS: Principal component and factor analyses suggested that each element, except care through the life cycle, contributed to the construct of primary care. Principal component analysis enabled ordering of specialties according to their "primary careness," suggesting that specialties other than family/general practice, pediatrics, and internal medicine make significant contributions to primary care. Factor analysis suggested that two factors related to process and content underlie the definition of primary care and emphasize the importance of integration of services provided. This analysis provides a basis for further empirical work to develop measures of primary care performance. CONCLUSIONS: National surveys need to be modified to provide a more comprehensive assessment of primary care in the United States. PMID- 9219495 TI - Limitations of epidemiologically based needs assessment. The case of prostatectomy. AB - OBJECTIVES: The aim of this study was to make epidemiologically based estimates of the prevalent and incident "need" for prostatectomy for lower urinary tract symptoms, defined as the numbers of men who would both benefit from and want the operation. METHODS: The methods involved a consensus panel, a two-stage postal survey of 1,480 men aged 55 years or older from eight general practices to the northwest of London, United Kingdom, and a multistate life table. RESULTS: The overall response rate was 69% (initial survey: 78%, follow-up survey: 88%). A trial-based estimate of number of candidates for prostatectomy (men with symptoms that were at least moderately severe and bothersome and who would probably or definitely want surgery) was 610 men in a population of 250,000. The corresponding incidence estimate (including men with symptoms recurring after spontaneous remission or surgery) was approximately 200 per year, including approximately 110 new cases. Consensus-based estimation, including categories of patients who have not yet been subject to a trial, gave much higher figures of approximately 3,000, 650, and 200 candidates, respectively. Adding the number of men who said they were "inclined to" choose surgery would almost double these figures. CONCLUSIONS: Estimates of need were highly sensitive to choice of indications and assumptions about patients' attitudes toward surgery. Population needs assessment for specific procedures will always involve judgment as well as epidemiological data and modeling. PMID- 9219496 TI - Mortality, hospital admissions, and medical costs of end-stage renal disease in the United States and Manitoba, Canada. AB - OBJECTIVES: National registry data suggest that mortality rates among patients with end-stage renal disease are lower in Canada than in the United States. Casemix and treatment variables, although limited in such instances, do not explain this difference. Using a more complete set of casemix and treatment variables from clinical databases, this study assesses mortality, hospital admission, and the cost of medical care for patients with end-stage renal disease treated in Manitoba, Canada and the United States. METHODS: Mortality rates were compared in patients with end-stage renal disease treated in the Province of Manitoba and a random sample of US patients enrolled in the US Renal Data System Casemix Severity Study. Hospital admission rates and costs of care were compared in Manitoba patients and in patients with end-stage renal disease in a large health care organization in Detroit, Michigan. RESULTS: Levels of serum creatinine, urea, and estimated glomerular filtration rate indicated more severe renal impairment at the outset of treatment in Manitoba than in the United States. Manitoba patients were more than twice as likely to receive kidney transplants as US Renal Data System patients. No patients in Manitoba used reprocessed dialyzers, compared with 57% of US Renal Data System patients. After adjustment for all casemix and treatment variables, the mortality rate was 47% higher in the United States. The hospital admission rate in Detroit was 41% lower than the hospital admission rate in Manitoba, which primarily reflects the doubled rate of transplantation in Manitoba. Adjusted total monthly costs were $503 higher in Detroit than in Manitoba. CONCLUSIONS: The higher mortality rates in the United States cannot be fully explained by adjustments for observable casemix or treatment variables. Further research is needed to identify factors that explain how Manitoba achieves a lower mortality rate while paying less for end-stage renal disease care than the United States. PMID- 9219497 TI - Analysis of the variety in surgeons' decision strategies for the management of left colonic emergencies. AB - OBJECTIVES: The aim of this study is to analyze surgeons' decision strategies about the optimal treatment for acute sigmoid resection for different patients. In particular, the authors wished to determine the predominant accepted treatment choice among surgeons, to determine the importance of patient characteristics for surgeons' evaluations of the appropriateness of treatments, and to identify the variety in decision strategies. METHODS: A survey was carried out among all surgical members of the Netherlands Society of Gastro Intestinal Surgery, who evaluated 16 patient cases. Approximately 70% of the members completed the survey. RESULTS: Overall, the predominant accepted strategy is resection, delayed anastomosis, and colostomy (Hartmann procedure). Consensus in terms of preferred treatment, however, was low. The most important factors influencing surgeons' evaluations of the appropriateness of treatments were the age of a patient, the degree of peritonitis, and the degree of fecal contamination. Further analysis showed that the variety in surgeons' decision strategies could not be explained by differences in experience, but was shown to be related to the evaluation of the appropriateness of treatment for 60-year-old patients and patients with a local peritonitis. Except for these factors, surgeons did not differ fundamentally in the evaluation of the factors that make a treatment more appropriate. Surgeons agreed about the optimal treatment for older patients in poor condition, although there is no epidemiologic literature to support this consensus position. CONCLUSIONS: This study showed that lack of consensus in surgeons' choice of treatment could be explained partly by disagreement of the appropriateness of treatments for some, rather than all, patients. PMID- 9219499 TI - A new instrument to measure patient satisfaction with mammography. Validity, reliability, and discriminatory power. AB - OBJECTIVES: The benefit of mammography depends on repeated use. Therefore, surveying the mammographic quality as judged by the users addresses an important topic. The authors assess the practicality, validity, reliability, and discriminatory power of a new, brief, multidimensional questionnaire for measuring patient satisfaction with mammography. Items measuring discomfort and attitudes toward repeat adherence were included. METHODS: A self-administered questionnaire was given to women from six radiology departments in Norway. Four hundred eighty-eight out of 550 women referred for screening or diagnostic mammography were included. Seventy-seven patients also completed the test/retest study, and 44 women additionally completed an Australian questionnaire. Scores for patient satisfaction on the structure, process, discomfort, and general satisfaction scales of the questionnaire were used as the main outcome measures. RESULTS: Response rate was 89%, and rate of completion was more than 95%. Strict psychometric criteria for construct validity and reliability were satisfied. Because lower levels of satisfaction were detectable with the new questionnaire but not with the Australian questionnaire and because an acceptable degree of variability in response was detected, support for discriminatory power was found. CONCLUSIONS: The discomfort dimension contributed substantially to validity and discriminatory power. Patient behavior with time may be monitored with the new questionnaire, thus representing a valuable tool for scientific and practical purposes. PMID- 9219498 TI - The relationship of patient satisfaction with care and clinical outcomes. AB - OBJECTIVES: The authors examine the relationship between three dimensions of patient satisfaction (quality of care, hospital care, and physician time) and two ways of looking at outcomes: absolute (status at 6 months after surgery) and relative (difference between baseline and follow-up status). METHODS: A total of 2,116 patients undergoing cholecystectomy were interviewed before surgery and again at 6 months. The baseline interview addressed health status (general functioning and specific symptoms) and risk factors. The follow-up interview included health status and a series of satisfaction questions. Outcomes included both overall health status and specific symptoms. Potential confounding factors, in addition to baseline status, such as demographics, casemix, and procedure type, were accounted for in the analysis. RESULTS: Each of the outcomes was related significantly to each of the satisfaction scales; however, the relative outcomes were related more strongly to satisfaction than were the absolute versions. Although the regression coefficients were highly significant, none of the outcomes measures accounted for more than 8% of the explained variance in the several satisfaction scores. CONCLUSIONS: Although outcomes and satisfaction are related, more goes into satisfaction than just outcomes. When determining their satisfaction with the care they have received, patients are more likely to focus on their present state of health than to consider the extent of improvement they have enjoyed. PMID- 9219500 TI - Outcomes of hypertension care. Simple measures are not that simple. PMID- 9219501 TI - Antibody-directed enzyme prodrug therapy: pharmacokinetics and plasma levels of prodrug and drug in a phase I clinical trial. AB - Antibody-directed enzyme prodrug therapy (ADEPT) was administered to ten patients in a phase I clinical trial. The aim was to measure plasma levels of the prodrug 4-[(2-chloroethyl)(2-mesyloxyethyl) amino] benzoyl-L-glutamic acid (CMDA) and the bifunctional alkylating drug (CJS11) released from it by the action of tumour localised carboxypeptidase G2 (CPG2) enzyme. New techniques were developed to extract the prodrug and drug from plasma by solid-phase absorption and elution and to measure CPG2 activity in plasma and tissue. All extracts were analysed by high-performance liquid chromatography (HPLC) and liquid chromatography-mass spectrometry (LC-MS). CPG2 activity was found in metastatic tumour biopsies but not in normal tissue, indicating that localisation had been successful. The clearing agent SB43-gal, given at 46.5 mg/m2, achieved the aim of clearing non tumour-localised enzyme in the circulation, indicating that conversion of prodrug to drug could take place only at the site of localised conjugate. Plasma prodrug did not always remain above its required threshold of 3 microM for the "therapeutic window" of 120 min after dosing, but the presence of residual prodrug after the first administration of each day indicated that this could be achieved during the remaining four doses over the following 8 h. Despite considerable inter-patient prodrug plasma concentration variability, the elimination half-life of the prodrug was remarkably reproducible at 18 +/- 8 min. Rapid appearance of the drug in plasma indicated that successful conversion from the prodrug had taken place, but also undesirable leakback from the site of localisation into the bloodstream. However, drug plasma levels fell rapidly by at least 50% at between 10 and 60 min with a half-life of 36 +/- 14 min. Analysis of the plasma extracts by LC/MS indicated that this technique might be used to confirm qualitatively the presence of prodrug, drug and their metabolites. PMID- 9219502 TI - Pharmacokinetics and pharmacodynamics of MX2 hydrochloride in patients with advanced malignant disease. AB - The purpose of the present study was to investigate the pharmacokinetics and pharmacodynamics of the new morpholino anthracycline drug MX2. A total of 27 patients with advanced cancer participated in a dose-escalation study in the first cycle of treatment with drug given i.v. at doses of 10-50 mg/m2 (total dose 16.8-107.5 mg). The mean total systemic plasma clearance (CL) of MX2 was 2.98 +/- 1.68 l/min, the mean volume of distribution at steady state was 1460 +/- 749 l and mean elimination half-life was 10.8 +/- 5.1 h. The area under the plasma concentration-time curve (AUC) of MX2 was linearly related to the dose per kilogram and the dose per body surface area (r2 = 0.43, P < 0.01 and r2 = 0.44, P < 0.01, respectively). CL did not correlate with total body weight, lean body mass or body surface area. The mean elimination half-lives of the metabolites M1, M2, M3 and M4 were 11.8 +/- 5.0, 21.9 +/- 11.8, 19.0 +/- 11.3 and 12.3 +/- 6.3 h, respectively. The fractional Emax model produced a much better fit to the relative nadir neutrophil count versus dose data (r2 = 0.42) than to the relative nadir neutrophil count versus AUC or peak concentration (Cmax) data (r2 = 0.15 and 0.09, respectively). There seemed to be a threshold dose of about 65 mg of MX2 at or above which a large proportion of patients had a nadir neutrophil count of less than 0.5 x 10(9)/l. This study shows that the pharmacokinetics of MX2 are similar to those of other anthracyclines. With other anthracyclines the degree of myelosuppression seems to depend more on the AUC and Cmax than on the delivered dose; however, with MX2 the degree of myelosuppression depends more on the dose given than on drug exposure expressed as the AUC or Cmax. PMID- 9219503 TI - Human tumor models in the severe combined immune deficient (scid) mouse. AB - PURPOSE: To test a number of established human tumor cell lines and early passage breast cancer (UACC2150) and melanoma cells (UACC1273) for growth in the scid mouse and the tumors' response to conventional chemotherapeutic drugs. METHODS: Established melanoma (A375, C81-61), colon (SW480), lung (A549), lymphomoblastoid leukemia (LCL-B), promyelocytic leukemia (HL60), prostate (PC-3, DU145), and breast (MCF7) cell lines were injected at subcutaneous (s.c.), intraperitoneal (i.p.), or mammary fat pad (MFP) sites. Tumor volume growth curves and survival curves were established for the various tumor cell lines. Carmustine (BCNU), cisplatin (CDDP), cyclophosphamide (CPA), doxorubicin, dacarbazine (DTIC), tamoxifen and vincristine were injected s.c. or i.p.. The chemotherapeutic drug effects on tumor volumes and survival were determined. RESULTS: Tumor growth occurred with each cell type. After i.p. injection, 90% mortality occurred within 26 to 60 days except for the early passage melanoma cell line UACC1273 with which mortality occurred within approximately 90 days. In the MCF7 breast model, treatment with tamoxifen (P < 0.001) and CPA (P < 0.0001) resulted in significant tumor growth delay compared with control groups. BCNU and CDDP resulted in significant tumor growth delays relative to control in SW480 colon cancer (P < 0.0014) and A375 melanoma (P < 0.0001) models, respectively. CPA and doxorubicin improved survival in the HL60 leukemia model (P = 0.0018). CONCLUSIONS: These scid mouse human tumor models appear to reflect the clinical situation in that clinically active chemotherapeutic drugs are similarly active in the scid mouse models. Therefore, the scid mouse models may be useful for testing new chemotherapeutic agents against various human cancer types. PMID- 9219505 TI - Phase II study of pyrazoloacridine in patients with advanced colorectal carcinoma. AB - PURPOSE: Pyrazoloacridine (PZA) is an acridine derivative selected for clinical development because of broad preclinical antitumor activity and solid tumor selectivity. Phase I evaluations with PZA have demonstrated predictable toxicity and suggested clinical efficacy. A phase II trial in patients with previously untreated advanced colorectal cancer was conducted. METHODS: PZA was administered at a dose of 750 mg/m2 intravenously over 3 h every 21 days to patients who received a total of 31 courses of PZA. RESULTS: In 15 patients evaluable for response, no responses were observed (0% response rate, 95% confidence interval 0 22%). Toxicity to PZA consisted of myelosuppression and neurotoxicity that was treatment-limiting in several instances. CONCLUSION: PZA at this dose and schedule of administration is inactive in patients with colorectal cancer. PMID- 9219504 TI - Multienzyme-mediated stable and transient multidrug resistance and collateral sensitivity induced by xenobiotics. AB - BACKGROUND: Determinants of cellular sensitivity to anticancer drugs include enzymes that catalyze their biotransformation. Coordinated induction of some of these enzymes is known to be caused by a number of dietary constituents, environmental contaminants, pharmacological agents and other xenobiotics, e.g. 3 methylcholanthrene and catechol. Despite the potential for inducing simultaneous changes in tumor cell sensitivity to a wide range of drugs, scant attention has been paid to the impact that dietary constituents and other xenobiotics might have on the therapeutic outcome of cancer chemotherapy. PURPOSE: The aim of this investigation was to demonstrate the potential of xenobiotic-induced multienzyme mediated stable and transient multidrug resistance/collateral sensitivity in a model system. METHODS: Human breast adenocarcinoma MCF-7/0 cells and a stably oxazaphosphorine-resistant subline thereof, MCF-7/OAP, were grown in the presence of 3-methylcholanthrene (3 microM), catechol (30 microM), or vehicle for 5 days. Spectrophotometric and spectrofluorometric assays were used to quantify catalytic activities and thus cellular levels of cytosolic class 3 aldehyde dehydrogenase, glutathione S-transferase, DT-diaphorase, UDP-glucuronosyl transferase and cytochrome P450 1A1. A colony-forming assay was used to quantify cellular sensitivities to several anticancer drugs. RESULTS: Relative to their untreated counterparts, MCF-7/0 and MCF-7/OAP cells treated with 3-methylcholanthrene or catechol transiently expressed elevated levels of cytosolic class 3 aldehyde dehydrogenase, glutathione S-transferase, DT-diaphorase and UDP-glucuronosyl transferase, and were transiently, more resistant to mafosfamide, melphalan, and mitoxantrone, and more sensitive to EO9. Further, MCF-7/0 and MCF-7/OAP cells treated with 3-methylcholanthrene, but not those treated with catechol, transiently expressed elevated levels of cytochrome P450 1A1 and were transiently more sensitive to ellipticine. Relative to MCF-7/0 cells, MCF-7/OAP cells stably overexpressed all but cytochrome P450 1A1 and were stably, more resistant to mafosfamide, melphalan and mitoxantrone, and more sensitive to EO9. Inclusion of relatively specific inhibitors of, or alternative substrates for, the enzymes of interest during drug exposure negated the influence of enzyme overexpression on cellular sensitivities to these agents. Untreated, and 3-methylcholanthrene- or catechol-treated, MCF-7/0 and MCF-7/OAP cells were equisensitive to vincristine and nearly so to doxorubicin. CONCLUSIONS: Collectively, these experiments illustrate the potential for both stable and transient xenobiotic-induced multienzyme-mediated multidrug resistance/collateral sensitivity that, although also the result of a single event, is mechanistically different from, and pertains to a largely different group of anticancer agents than does, the multidrug resistance caused by cell surface multidrug transporters. PMID- 9219506 TI - Tubulin from paclitaxel-resistant cells as a probe for novel antimicrotubule agents. AB - PURPOSE: Treatment with paclitaxel (PTX) can lead to the appearance of drug resistance with accompanying changes in tubulin. The purpose of this study was to develop an assay for microtubule-active agents that are able to circumvent changes in tubulin that result in acquired resistance to paclitaxel. METHODS: The assay measured the promotion of microtubule polymerization when target agents were added to solutions containing tubulin purified from cultured cells. Tubulin was prepared from PTX-sensitive 1A9 ovarian carcinoma cells and from a PTX resistant clone. Polymerization was monitored spectrophotometrically and validated by electron microscopy. RESULTS: Exposure of tubulin isolated from PTX sensitive 1A9 ovarian carcinoma cells to substoichiometric PTX resulted in polymerization equivalent to that observed with brain tubulin. In contrast, tubulin from a PTX-resistant 1A9 clone failed to polymerize under identical conditions. If a C-2-modified analog of PTX (2-debenzoyl-2-(m azidobenzoyl)paclitaxel) was substituted for PTX in the same experiment, the tubulins from both sensitive and resistant cells polymerized as well as brain tubulin. As predicted from these results, the PTX analog was nearly as cytotoxic to the PTX-resistant cells as it was to the parental cells: the relative resistance of the resistant cells compared to the parental is only 3-5-fold for the PTX analog versus 25-30-fold for PTX. CONCLUSION: Polymerization of purified tubulin from the paclitaxel-resistant cells provided an assay for agents able to circumvent the tubulin alterations that result in acquired paclitaxel resistance. PMID- 9219507 TI - Dose intensity of uracil and tegafur in postoperative chemotherapy for patients with poorly differentiated gastric cancer. AB - A retrospective analysis of postoperative chemotherapy had shown the continuous administration of UFT, an oral preparation of 1-(2-tetrahydrofuryl)-5 fluorouracil (tegafur) and uracil at a molar ratio of 1:4, to be effective for poorly differentiated gastric cancer. We therefore sought to determine prospectively the effective dose of postoperative chemotherapy with UFT for patients with poorly differentiated gastric cancer following a curative resection. We determined the effect of the combined intravenous administration of mitomycin C (MMC) and oral treatment with protein-bound polysaccharide Kreha (PSK), extracted from the basidiomycete Coriolus versicolor, and UFT at a dose of either 8 mg/kg or 12 mg/kg daily for 1 year. A total of 224 patients with poorly differentiated stage II-IV gastric cancer were entered into this study after undergoing a curative resection. No differences were observed between the two treatment groups in terms of prognostic factors, the toxicity rate or the doses of the drugs prescribed, other than UFT. The higher dose of UFT in maintenance therapy led to a decrease in the recurrence rate (P < 0.05), and increases in disease-free survival and cause-specific survival (P < 0.05). UFT at 12 mg/kg in postoperative chemotherapy was thus found- to improve the postoperative results with no increase in toxicity for poorly differentiated gastric cancer, and is also cost-effective for outpatients. PMID- 9219508 TI - Antiestrogenic activities of alternate-substituted polychlorinated dibenzofurans in MCF-7 human breast cancer cells. AB - PURPOSE: 1,3,6,8-Substituted alkyl polychlorinated dibenzofurans (PCDFs), typified by 6-methyl-1,3,8-triCDF (MCDF), inhibit 17 beta-estradiol (E2)-induced responses in the rodent uterus and human breast cancer cells. The major purpose of the experiments reported here was to determine the structure-dependent antiestrogenic activities of several alternate-substituted (1,3,6,8- and 2,4,6,8 ) PCDFs. METHODS: The antiestrogenic activities were determined in MCF-7 human breast cancer cells using two assays, that is E2-induced cell proliferation and induction of chloramphenicol acetyl transferase (CAT) activity in cells transiently transfected with the E2-responsive Vit-CAT plasmid. RESULTS: MCDF (10(-5) M), 6-isopropyl-1,3,8-triCDF, 6-ethyl-1,3,8-triCDF, 3-isopropyl-6-methyl 1,8-diCDF, and 6-methyl-2,4,8-triCDF, inhibited both E2-induced cell proliferation and CAT activity in MCF-7 cells. All of the remaining ten congeners inhibited either E2-induced cell proliferation or CAT activity, but not both responses. CONCLUSIONS: The antiestrogenic activity of the alternate-substituted PCDFs involves interactions between the aryl hydrocarbon and estrogen receptor signaling pathways. Although these compounds exhibited antiestrogenic activity in MCF-7 cells, the effects of individual congeners were response-specific, and there were no apparent structure-activity relationships. PMID- 9219509 TI - Resistance to paclitaxel mediated by P-glycoprotein can be modulated by changes in the schedule of administration. AB - PURPOSE: Increasing use of paclitaxel in clinical oncology has stimulated interest in its mechanisms of resistance and ways to overcome these. Studies were performed with paclitaxel to determine the role of P-glycoprotein in drug sensitivity, and the effect of schedule on relative resistance. We have previously reported that prolonged exposure to P-glycoprotein substrates decreases relative resistance in multidrug resistant cells. METHODS: Using both unselected and drug-selected cell lines, cross-resistance and cytotoxicity reversal studies using cyclosporin A were performed. In multidrug-resistant cells, cross-resistance was evaluated after 3-, 24-, and 96-h exposures to paclitaxel. RESULTS: Cross-resistance to paclitaxel in P-glycoprotein-expressing sublines was shown to be comparable to that of other drugs transported by P glycoprotein. Sensitivity to paclitaxel could be modulated by cyclosporin A in unselected cell lines expressing P-glycoprotein and not in P-glycoprotein negative cell lines. Resistance to paclitaxel was reduced tenfold by increasing the duration of exposure in P-glycoprotein-expressing cells. This effect was not observed in a paclitaxel-resistant cell line which does not express P glycoprotein. CONCLUSIONS: These studies extend observations on the schedule dependence of paclitaxel cytotoxicity and the role of P-glycoprotein in mediating paclitaxel sensitivity. The schedule dependence of relative resistance suggests that infusional paclitaxel may help in overcoming P-glycoprotein-mediated resistance. PMID- 9219510 TI - Disposition of conjugate-bound and free doxorubicin in tumor-bearing mice following administration of a BR96-doxorubicin immunoconjugate (BMS 182248). AB - PURPOSE: The chimeric BR96-doxorubicin (DOX) immunoconjugate, BMS 182248, has induced remissions and cures of human lung adenocarcinoma (L2987) implanted in athymic mice. The purpose of this study was to evaluate the biodistribution of DOX after BMS 182248 administration to tumor-bearing mice and to evaluate the ability of BMS 182248 to target DOX to tumors. METHODS: For this evaluation, L2987-implanted mice were given BMS 182248 (5 mg DOX/kg; three doses 4 days apart) and the levels of both conjugate-bound and free DOX in plasma, tumor, liver and heart were determined. RESULTS: Conjugate-bound DOX comprised the majority of plasma DOX, with relatively low levels of free DOX present. From plasma, conjugate-bound DOX distributed to the tissues examined with the order of concentration (per gram of tissue) being tumor > liver > heart. Free DOX was also detected in liver and heart, but at concentrations lower than those present after an equivalent DOX dose (5 mg/kg; three doses 4 days apart). The total exposure of heart to free DOX after BMS 182248 administration was about one-quarter of that found after the administration of DOX alone. The elimination kinetics of both conjugate-bound and free DOX from heart and liver after BMS 182248 administration paralleled those observed from plasma, indicating that equilibrium had been attained between these nontumor tissues and plasma. The elimination kinetics of both entities from tumors, however, were different from those from plasma, liver and heart. BMS 182248 produced sustained levels of both conjugate-bound and free DOX which were present throughout the experiment. This suggested that, in contrast to normal tissues, tumor tissue retention of BMS 182248 by antigen promoted binding had occurred and that kinetics of free DOX in the tumors were controlled by the rate of release of DOX from tumor-associated BMS 182248. As a result of this retention, the tumor concentrations of free DOX after BMS 182248 administration exceeded those produced by i.v. administration of DOX at the same dose, a finding consistent with the greater antitumor activity of BMS 182248 relative to DOX. BMS 182248 also liberated DOX upon incubation with rat liver lysosomes and was accumulated by L2987 cells in culture, with the subsequent intracellular release of DOX. CONCLUSIONS: BMS 182248 effectively delivered DOX to L2987 xenografts implanted in athymic mice and produced higher and more prolonged tumor concentrations of free DOX than the administration of DOX alone. Following BMS 182248 administration, normal tissues (liver and heart) were exposed to lower overall concentrations of free DOX than were produced by administration of an equivalent DOX dose. PMID- 9219511 TI - Disposition of irinotecan and SN-38 following oral and intravenous irinotecan dosing in mice. AB - The present study was conducted to quantitate the disposition of irinotecan lactone and its active metabolite SN-38 lactone in mice following oral and intravenous administration, and to evaluate the systemic exposure of irinotecan lactone and SN-38 lactone associated with antitumor doses of irinotecan lactone in mice bearing human tumor xenografts. Nontumor-bearing mice were given a single oral or intravenous irinotecan dose (5, 10, 40, or 75 mg/kg), and serial plasma samples were subsequently obtained. Irinotecan and SN-38 lactone plasma concentrations were measured using an isocratic HPLC assay with fluorescence detection. The disposition of intravenous irinotecan lactone was modeled using a two-compartment pharmacokinetic model, and the disposition of oral irinotecan and SN-38 lactone was modeled with noncompartmental methods. Irinotecan lactone showed biphasic plasma disposition following intravenous dosing with a terminal half-life ranging between 1.1 to 3 h. Irinotecan lactone disposition was linear at lower doses (5 and 10 mg/kg), but at 40 mg/kg irinotecan lactone clearance decreased and a nonlinear increase in irinotecan lactone AUC was observed. The steady-state volume of distribution ranged from 19.1 to 48.1 l/m2. After oral dosing, peak irinotecan and SN-38 lactone concentrations occurred within 1 h, and the irinotecan lactone bioavailability was 0.12 at 10 mg/kg and 0.21 at 40 mg/kg. The percent unbound SN-38 lactone in murine plasma at 1000 ng/ml was 3.4 +/- 0.67%, whereas at 100 ng/ml the percent unbound was 1.18 +/- 0.14%. Irinotecan and SN-38 lactone AUCs in micebearing human neuroblastoma xenografts were greater than in nontumor-bearing animals. Systemic exposure to unbound SN-38 lactone in nontumor-bearing animals after a single oral irinotecan dose of 40, 10, and 5 mg/kg was 28.3, 8.6, and 2.9 ng h/ml, respectively. Data from the present study provide important information for the design of phase I studies of oral irinotecan. PMID- 9219512 TI - Effect of single and multiple administration of an O6-benzylguanine/temozolomide combination: an evaluation in a human melanoma xenograft model. AB - The purpose of the present study was to examine the effect of O6-benzylguanine (O6-BG) on the antitumour activity and toxicity of 8-carbamoyl-3-methylimidazo [5, 1-d]-1,2,3,5-tetrazine-4(3H)-one (temozolomide) in a human malignant melanoma xenograft model following single and multiple administration of the combination. O6-BG irreversibly inactivates the DNA-repair protein O6-alkylguanine-DNA alkyltransferase (AGT), which confers resistance to temozolomide. Preadministration of O6-BG (35 mg/kg, i.p.) 1 h prior to temozolomide (i.p.) was examined using single and daily x5 dosing regimens in athymic mice bearing subcutaneous A375P xenografts. The AGT activity of A375P tumors was 95 +/- 8 fmol/mg protein (mean +/- SE, n = 4). O6-BG alone completely suppressed xenograft AGT activity within 1 h of administration but had no effect upon tumor growth. O6 BG did not significantly increase the tumor growth delay induced by a single 200 mg/ kg dose of temozolomide (P > 0.05, two-tailed Mann-Whitney test) but did increase the associated mean body weight loss (P < 0.025). In contrast, when the same dose of temozolomide was divided into five equal fractions (40 mg/kg) and given with O6-BG on 5 consecutive days, a comparable increase in toxicity was accompanied by a very significant increase in tumor growth delay (P < 0.0025), equivalent to that produced by a 3-fold greater dose of temozolomide alone. O6-BG with temozolomide also produced a greater antitumour effect than an equitoxic dose of temozolomide alone on this schedule (P < 0.005). These data indicate that the enhancement of temozolomide antitumour activity by O6-BG preadministration is dependent upon the schedule of drug administration, with multiple dosing of O6-BG + temozolomide producing the greatest effect. The results also suggest that prolonged administration of the combination can lead to an increase in the therapeutic index of temozolomide. PMID- 9219513 TI - Feasibility trial of high-dose 7-day continuous-infusion ifosfamide given on an outpatient basis. AB - High-dose ifosfamide (HD-IFX) has shown significant antitumor activity in advanced sarcoma and breast carcinoma. The use of uroprotective agents and the availability of ambulatory continuous-infusion pumps has allowed dose escalation in the administration of ifosfamide (IFX) on an outpatient schedule. We report the results of a phase II trial of IFX given at high doses to heavily pretreated patients. IFX was infused at 2 g/m2 per day for a total of 7 days through a central venous access, with cycles being repeated every 21 days. Mesna was given concomitantly at equimolar doses. No hematopoietic support was used. A total of 27 heavily pretreated patients whose disease had progressed during conventional dose chemotherapy were included (14 sarcomas, 10 breast carcinomas, and 3 bladder carcinomas). Reversible neutropenia and gastrointestinal toxicity were the most frequently encountered toxicities. Only two patients developed transient renal failure, and two others developed central nervous system toxicity. No treatment related death was observed. Of 22 patients who were evaluable for response, 6 (27%) showed an objective response (OR), all ORs being partial responses (PRs) with a median duration of 6 months, and 12 patients had stable disease (SD; 55%) with a median duration of 3.5 months. The median overall survival (OS) was 6 months. Three patients underwent high-dose chemotherapy after showing a response to our IFX schedule. We conclude that continuous-infusion IFX given in an outpatient setting is a feasible and active regimen that produces, a manageable toxicity profile in heavily pretreated breast cancer and sarcoma patients. Early institution of this schedule in less advanced stages could improve the results obtained. PMID- 9219514 TI - The retinoblastoma protein: a master regulator of cell cycle, differentiation and apoptosis. AB - The retinoblastoma susceptibility gene is a tumour suppressor and its product retinoblastoma protein (pRb) has been known for 10 years as a repressor of progression towards S phase. Its major activity was supposed to be sequestration or inactivation of the transcription factor E2F which is required for activation of S phase genes. However, within recent years growing evidence has been accumulating for a more general function of pRb at both the transcriptional level and the cellular level. pRb not only regulates the activity of certain protein encoding genes but also the activity of RNA polymerase pol I and pol III transcription. This protein appears to be the major player in a regulatory circuit in the late G1 phase, the so-called restriction point. Moreover, it is involved in regulating an elusive switch point between cell cycle, differentiation and apoptosis. Here, it seems to cooperate with another major tumour suppressor, p53. Thus, pRb sits at the interface of the most important cell-regulatory processes and therefore deserves close attention by specialists from different fields of research. This review provides an introduction to the complex functions of pRb. PMID- 9219516 TI - Structural and functional domains of the troponin complex revealed by limited digestion. AB - Troponin (Tn), consisting of three subunits, TnT, TnC, and TnI, plays a crucial role in the calcium-dependent regulation of vertebrate striated muscle contraction. In the present study, we have applied limited proteolysis to the Tn complex in order to study domain structures and to detect conformational differences of Tn under different conditions. We found that both TnT and TnI were susceptible to chymotryptic digestion: while TnT was cleaved into TnT-(1-158) peptide and TnT-(159-259)-peptide irrespective of Ca2+ concentration, the cleavage sites of TnI were dependent on the Ca2+ occupancy of TnC. In addition, we characterized the effects of depletion of the C-terminal part of TnI on acto S1 ATPase activity. The TnT-(159-259)-peptide-TnC-TnICa-frag complex [TnICa-frag = (TnI-(1-134 and 1-140)-peptide], which was produced in the presence of CaCl2 and MgCl2, retains both the activating and inhibitory capabilities of whole Tn on the acto-S1 ATPase activity, while TnT-(159-259)-peptide-TnC-TnIMg-frag complex [TnIMg-frag = (TnI-(1-116)-peptide], which was obtained in the presence of MgCl2 and EGTA, lost its ability to activate acto-S1 ATPase activity. Our results indicate that residues 117-134 or 117-140 of TnI undergo structural changes upon Ca(2+)-binding to the regulatory sites of TnC and are necessary for the Ca(2+) dependent inhibitory action of the Tn complex on acto-S1 ATPase activity. We also showed that residues 135-181 or 141-181 of TnI are involved in the interaction of Tn with the tropomyosin-actin filament. PMID- 9219517 TI - Expression studies on the ba3 quinol oxidase from Paracoccus denitrificans. A bb3 variant is enzymatically inactive. AB - Expression of the quinol oxidase from Paracoccus denitrificans has been examined using a polyclonal antibody directed against subunit II and a promoter probe vector carrying the promoter region of the qox operon. Under aerobic conditions nitrate and nitrite act as specific inducers of the expression. To obtain an enzymatically competent quinol oxidase complex, an intact ctaB gene is required, which constitutes part of the cta operon coding for the aa3 cytochrome c oxidase of P. denitrificans. Deletion of ctaB leads to a change in heme composition of the quinol oxidase with heme b replacing the high-spin heme a of the binuclear center, causing loss of electron transport activity. PMID- 9219515 TI - Association of calmodulin with nuclear structures in starfish oocytes and its role in the resumption of meiosis. AB - The resumption of meiosis in prophase-arrested starfish oocytes is induced by the hormone 1-methyladenine, which has been shown previously to induce a calcium transient in the nucleus which at this stage is called the germinal vesicle. This transient precedes the breakdown of the germinal vesicle (GVBD). Experiments were performed to establish whether nuclear calmodulin (CaM) was involved in the progression of the meiotic cycle. CaM antagonists, antibodies, and an inhibitory peptide corresponding to the CaM-binding domain of myosin-light-chain kinase have been injected into the nucleus of prophase-arrested starfish oocytes. The antagonists failed to affect the final response to 1-methyladenine, i.e. GVBD, although two antagonists delayed it, whereas the peptide inhibitor and the antibodies completely inhibited it. The antibodies suppressed the nuclear Ca2+ spikes that were shown by previous work to be induced by the photoreleasing of caged adenosine 3',5'-(cyclic)diphosphate ribose in the germinal vesicle. Immunofluorescence staining of isolated starfish oocyte nuclei with CaM antibodies showed CaM in the envelope and in the nucleolus. Immunogold labelling of oocytes revealed aggregates of CaM and of a 36-kDa protein, of the heterogeneous ribonucleoprotein particles (hnRNP), in electron-dense hnRNP in the nuclear matrix. 1-Methyladenine induced the disappearance of these hnRNP from the nucleoplasm and the translocation of CaM and the 36-kDa protein previously associated with them to the cytoplasm, prior to the breakdown of the nuclear envelope. PMID- 9219519 TI - Identification of phosphopeptide ligands for the Src-homology 2 (SH2) domain of Grb2 by phage display. AB - We report here on the identification of phosphopetide ligands which interact with the Src-homology 2 (SH2) domain of the adapter protein Grb2 by screening a random peptide library established on phage. Phage were phosphorylated in vitro at an invariant tyrosine residue by a mixture of phosphotyrosine kinases c-Src, Blk and Syk. Selection of binding motifs was carried out by interaction of the library with the recombinant SH2 domain of Grb2 expressed as a glutathione S-transferase (GST) fusion protein. Several subsequent cycles of selection led to the enrichment of phage which bound to the GST-Grb2 SH2 domain only when previously phosphorylated. Sequence analysis revealed that all of the selected phage displayed peptides with the consensus motif Y*M/ENW (Y* denotes phosphotyrosine). One of these peptides, bearing the Y*ENW motif, bound the Grb2 SH2 domain with a threefold higher affinity than the peptide motif Y*VNV derived from the natural ligand Shc. Thus, phage display can be employed to rapidly identify high affinity ligands to SH2 domains. PMID- 9219518 TI - Characterization of potential antagonists of human interleukin 5 demonstrates their cross-reactivity with receptors for interleukin 3 and granulocyte macrophage colony-stimulating factor. AB - The ligand-binding alpha-chain of the human interleukin 5 (IL-5) receptor was expressed in its soluble form, lacking the transmembrane and cytoplasmic domains, from recombinant baculovirus. The soluble receptor was used in a scintillation proximity assay to identify two chemical compounds that inhibit binding of human IL-5 to the soluble receptor alpha chain with IC50 of 8 microM and 11 microM. These compounds also inhibited the interaction of human IL-5 with its membrane bound receptor, composed of the ligand-binding alpha chain and signal-transducing beta chain, and prevented signaling through the receptor. Analysis by surface plasmon resonance and matrix-assisted laser-desorption/ionization mass spectrometry showed that the identified compounds bound irreversibly to the receptor at a 1:1 (mol/mol) ratio, suggesting a covalent interaction with the alpha chain of the human IL-5 receptor. Both compounds also inhibited the interaction of the receptors for interleukin 3 (IL-3) and granulocyte-macrophage colony-stimulating factor (GM-CSF), which are involved in hematopoietic differentiation and activation of immune cells, thus eliminating them as potential therapeutic agents. The inhibition of the structurally closely related receptors for IL-5, IL-3 and GM-CSF by both compounds, while binding of interleukin-4 to its receptor was not affected, suggests that a similar reactive site exists in the ligand-binding domains of the receptors for IL-5, IL-3 and GM CSF. PMID- 9219520 TI - 13C-NMR studies of transmembrane electron transfer to extracellular ferricyanide in human erythrocytes. AB - Human erythrocytes are known to reduce ferricyanide (hexacyanoferrate) [Fe(CN)6]3 to ferrocyanide [Fe(CN)6]2- in an extracellular reaction that involves the transmembrane transfer of reducing equivalents; potentially these could be either electrons from NADH, formed in glycolysis inside the cells or transmembrane exchange of reduced solutes. The 13C-NMR resonance of [Fe(13CN)6]3- (which was synthesised in our laboratory) was seen to be very broad while that of ferrocyanide was narrow. This phenomenon formed the basis of a simple non invasive procedure to study ferricyanide reduction in high-haematocrit suspensions of erythrocytes. The method should be directly applicable to other cell types. In a series of experiments, erythrocyte metabolism was studied in the presence of ferricyanide, using 1H, 13C, and 31P NMR spectroscopy. Incubating the cells with 13C-labelled glucose enabled the rate of ferricyanide reduction, glucose utilisation, and lactate and bicarbonate production to be measured simultaneously. Various metabolic states were imposed as follows: glycolysis was inhibited with F- and iodoacetate; glucose transport was inhibited with phloretin and cytochalasin B; and anion transport was inhibited with dinitrostilbene 2,2' disulfonate and p-chloromercuriphenyl sulfonate. Earlier work was confirmed, showing that ascorbate is intimately involved in the reduction reaction; but its main action appears not to be mediated by membrane transport but in a membrane associated redox-protein complex that is functionally linked to glycolysis. Also, large differences (factors of three) in the rate of the reduction reaction were recorded in erythrocytes from different, apparently healthy, donors. PMID- 9219521 TI - Structure and function of a second gene cluster encoding the formate dehydrogenase of Wolinella succinogenes. AB - Wolinella succinogenes contains a single formate dehydrogenase, but two gene loci (fdhI and fdhII) code for the subunits of the enzyme. The nucleotide sequence of fdhII is almost identical with that of fdhI in the region comprising fdhEABCD. The sequences of fdhI and fdhII differ in the promotor regions upstream of fdhE. Deletion mutants lacking either fdhI or fdhII synthesize functional formate dehydrogenases, as shown by growth with formate as electron donor and either fumarate or polysulfide as acceptor substrates, and by the presence of the FdhA subunit and of enzyme activity. In the wild-type strain, the fdhI genes appear to be expressed preferentially during growth with formate and fumarate. The six times greater amount of the enzyme present upon growth with formate and polysulfide is due to the expression of both fdhI and fdhII. The transcription start sites were located 196-bp and 129-bp upstream of the fdhE start codons of fdhI and fdhII, respectively. An apparently single transcript (5.6 kbp) was detected in polysulfide-grown W. succinogenes by Northern-blot analysis, suggesting that the five open reading frames form operons. PMID- 9219522 TI - Aminopeptidase N from Bombyx mori as a candidate for the receptor of Bacillus thuringiensis Cry1Aa toxin. AB - Cry1Aa toxin-binding proteins from the midgut brush border membrane vesicles of Bombyx mori, a toxin-susceptible silkworm, were analyzed to find candidates for the toxin receptors. Ligand blotting showed that Cry1Aa toxin bound to a 120-kDa protein. A part of the 120-kDa protein was solubilized from the membrane vesicles with phosphatidylinositol-specific phospholipase C, resulting in a 110-kDa protein which therefore may be linked to a glycosyl-phosphatidylinositol anchor. The 120-kDa and 110-kDa Cry1Aa toxin-binding proteins were solubilized with detergent or pohosphatidylinositol-specific phospholipase C, respectively, and purified using anion-exchange chromatography. Scatchard plot analysis for the specific binding of purified 110-kDa protein to Cry1Aa toxin yielded a Kd value of 7.6 nM, which was similar to that for the binding of intact brush border membrane vesicles to the toxin. N-terminal and internal amino acid sequences of the 120-kDa and 110-kDa proteins showed high degrees of similarity to those of aminopeptidase N, a putative Cry1Ac toxin receptor, reported in Manduca sexta and Heliothis virescens. On this basis, the 120-kDa Cry1Aa toxin-binding protein from B. mori was identified as a member of the aminopeptidase family. PMID- 9219523 TI - Spectroscopic probes of the individual and combined effects of Triton X-100 and chloroform on serum albumins and serum-albumin.bilirubin complexes. AB - The effects of the non-ionic surfactant Triton X-100 on the biphasic induced CD spectra of bilirubin complexes of human and bovine serum albumins (HSA and BSA) are divergent. While Triton X-100 inverts the induced CD spectrum of HSA.bilirubin, this surfactant enhances the ellipticity values of induced CD of BSA.bilirubin without inversion. The effect of Triton X-100 on the characteristic ultraviolet-CD spectra of the albumins are similar; both the albumins are denatured from their native globular structures. The anionic surfactant SDS, unlike non-ionic Triton X-100, dislodges the ligand from its protein complexes, indicating that both electrostatic and hydrophobic forces are involved in binding of bilirubin to the albumins. The aprotic solvent chloroform inverts the biphasic induced CD spectra of HSA.bilirubin and BSA.bilirubin, whereas CHCl3 has relatively little effect on the ultraviolet CD spectra of the albumins. The combined effect of Triton X-100 and CHCl3 shows that the effect of CHCl3 predominates over that of Triton X-100. The perturbing effects of Triton X-100 and CHCl3 on the CD or induced CD spectra of the proteins or their bilirubin complexes are reversible, and independent of the order in which components were added. The observations suggest that the denaturation of the albumins by Triton X 100 or solvation of CHCl3 within albumins markedly alter the internal topography or dynamics of the receptor sites, triggering alterations of the chirality of the bound pigment in sign and/or magnitude. PMID- 9219524 TI - A determination of the solution conformation of secretoneurin, a neuropeptide originating from the processing of secretogranin II, by 1H-NMR and restrained molecular dynamics. AB - Secretoneurin is a 33-amino-acid polypeptide generated by proteolytic cleavage of secretogranin II at paired dibasic sequences. It has recently been shown that secretoneurin exerts biological activities such as stimulation of dopamine release from striatal neurons and activation of monocyte migration, suggesting that the peptide may modulate both neurotransmission and inflammatory response. In the present study, we have investigated the conformation of synthetic secretoneurin in methanol solution by two-dimensional 1H-NMR, circular dichroism and molecular modeling. Using sequential information, specific assignments have been made for resonances arising from all protons, except for the labile proton of the N-terminal Thr of the peptide. The solution structure of secretoneurin has been determined by distance geometry and restrained molecular dynamics, using distance and dihedral constraints derived from the NMR data. The conformation obtained is composed of two contiguous alpha-helices comprising residues Glu3 Gln8 and Pro11-Gly25. An excellent concordance was observed between these conformational data and prediction with the AGADIR program for the location for the helices in the sequence. These conformational data should help to elucidate the involvement of the tertiary interactions and to design secretoneurin analogs. PMID- 9219525 TI - Intermediates in the assembly of bacteriorhodopsin investigated by time-resolved absorption spectroscopy. AB - The in vitro folding and assembly kinetics of bacteriorhodopsin have been studied by absorption spectroscopy. Folding is initiated by rapid stopped-flow mixing of denatured apoprotein (bacterio-opsin) in SDS micelles and mixed dimyristoylglycerophosphocholine/Chaps micelles containing retinal. The apparent mixing rate of the two types of micelles has been determined by time-resolving the changes in light scattering by the micelles. Micelle mixing appears to occur in two stages: a fast phase with an apparent rate constant of about 420 s-1, and a second phase with an apparent rate constant of about 10 s-1. A rate constant of similar magnitude to the latter has previously been assigned to a protein-folding event on the basis of protein fluorescence studies [Booth, P. J., Farooq, A. & Flitsch, S. L. (1996) Biochemistry 35, 5902-5909]. However the results presented here show that this rate constant may be associated with a rearrangement of the mixed detergent/lipid micelles. When the changes in the retinal absorption band are time-resolved during assembly of bacteriorhodopsin, a retinal-protein intermediate, with an absorption maximum of about 430 nm, has been identified. This absorption maximum lies between that of unbound retinal (at about 380 nm) and the native chromophore (at about 560 nm). A comparison of fluorescence and absorption data, together with previous evidence [Booth, P. J., Flitsch, S. L., Stern, L. J., Greenhalgh, D. A., Kim, P. S., & Khorana, H. G. (1995) Nat. Struct. Biol. 2, 139-143], suggests that the covalent Schiff-base link to retinal is not formed in the 430-nm-absorbing intermediate. PMID- 9219526 TI - Man9-mannosidase from pig liver is a type-II membrane protein that resides in the endoplasmic reticulum. cDNA cloning and expression of the enzyme in COS 1 cells. AB - Man9-mannosidase, one of three different alpha 1,2-exo-mannosidases known to be involved in N-linked oligosaccharide processing, has been cloned in lambda gt10, using a mixed-primed pig liver cDNA library. Three clones were isolated which allowed the reconstruction of a 2731-bp full-length cDNA. The cDNA construct contained a single open reading frame of 1977 bp, encoding a 659-residue polypeptide with a molecular mass of approximately 73 kDa. The Man9-mannosidase specificity of the cDNA construct was verified by the observation that all peptide sequences derived from a previously purified, catalytically active 49-kDa fragment were found within the coding region. The N-terminus of the 49-kDa fragment aligns with amino acid 175 of the translated cDNA, indicating that the catalytic activity is associated with the C-terminus. Transfection of COS 1 cells with the Man9-mannosidase cDNA gave rise to a > 30-fold over-expression of a 73 kDa protein whose catalytic properties, including substrate specificity, susceptibility towards alpha-mannosidase inhibitors and metal ion requirements, were similar to those of the 49-kDa enzyme fragment. Thus deletion of 174 N terminal amino acids in the 73-kDa protein appears to have only marginal influence on the catalytic properties. Structural and hydrophobicity analysis of the coding region, as well as the results from tryptic degradation studies, point to pig liver Man9-mannosidase being a non-glycosylated type-II transmembrane protein. This protein contains a 48-residue cytosolic tail followed by a 22 residue membrane anchor (which probably functions as internal and non-cleavable signal sequence), a lumenal approximately 100-residue-stem region and a large 49 kDa C-terminal catalytic domain. As shown by immuno-fluorescence microscopy, the pig liver enzyme expressed in COS 1 cells, is resident in the endoplasmic reticulum, in contrast to COS 1 Man9-mannosidase from human kidney which is Golgi located [Bieberich, E. & Bause, E. (1995) Eur. J. Biochem. 233, 644-649]. Localization of the porcine enzyme in the endoplasmic reticulum is consistent with immuno-electron-microscopic studies using pig hepatocytes. The different intracellular distribution of pig liver and human kidney Man9-mannosidase is, therefore, enzyme-specific rather than a COS-1-cell-typical phenomenon. Since we observe approximately 81% sequence similarity between the two alpha-mannosidases, we deduce that the localization in either endoplasmic reticulum or Golgi is likely to be sequence-dependent. PMID- 9219527 TI - The product of the molybdenum cofactor gene mobB of Escherichia coli is a GTP binding protein. AB - The mob mutants of Escherichia coli are pleiotropically defective in molybdoenzyme activities because they are unable to catalyse the conversion of molybdopterin guanine dinucleotide, the active form of the molybdenum cofactor. The mob locus comprises two genes. The product of mobA, protein FA, has previously been purified to homogeneity and is able to restore molybdoenzyme activities following incubation with cell extracts of mob strains. The mobB gene, although not essential for the biosynthesis of active molybdoenzymes, encodes a protein which, sequence analysis strongly suggests, contains a nucleotide-binding site. We have overproduced the products of both the mobA and mobB genes in engineered E. coli strains and purified each to homogeneity. The preparation of protein FA (MobA) is simpler than that previously published and produces a much greater yield of active protein. The isolated MobB protein, which is dimeric in solution, acts in the presence of protein FA, to enhance the level of nitrate reductase activation achieved on incubation with mob cell extracts. Equilibrium dialysis experiments show that purified MobB binds 0.83 mol GTP/mol protein with a Kd of 2.0 microM. Isolated MobB also catalyses a low GTPase activity (turnover number of 3 x 10(-3) min-1) with a K(m) for GTP to GDP of 7.5 microM. Under the conditions tested, protein FA did not affect the GTP-binding or GTPase activity of MobB. Intrinsic (tryptophan) protein fluorescence measurements show that MobB also binds the nucleotides ATP, TTP and GDP, but with lower affinity than GTP. These results are consistent with a model whereby MobB binds the guanine nucleotide which is attached to molybdopterin during the biosynthesis of the molybdenum cofactor. PMID- 9219528 TI - Critical glutamic acid residues affecting the mechanism and nucleotide specificity of Vibrio harveyi aldehyde dehydrogenase. AB - Fatty aldehyde dehydrogenase (ALDH) from the luminescent marine bacterium, Vibrio harveyi, differs from other ALDHs in its unique specificity and high affinity for NADP+. Two glutamic acid residues, Glu253 and Glu377, which are highly conserved in ALDHs, were investigated in the present study. Mutation of Glu253 to Ala decreased the kcat for ALDH activity by over four orders of magnitude without a significant change in the K(m) values for substrates or the ability to interact with nucleotides. Both thioesterase activity and a pre-steady-state burst of NAD(P)H were also eliminated, implicating Glu253 in promoting the nucleophilicity of the cysteine residue(Cys289) involved in forming the thiohemiacetal intermediate in the enzyme mechanism. Mutation of Glu377 to Gln (E377Q mutant) selectively decreased the kcat for NAD(+)-dependent ALDH activity (> 10(2)-fold) compared to only a 6-fold loss in NADP(+)-dependent activity without comparable changes to the K(m) values for substrates. Consequently, the E377Q mutant had a very high specificity for NADP+(kcat/K(m) > 10(3) of that for NAD+) which was over 20 times higher than that of the wild-type ALDH. Although a pre-steady-state burst of NAD(P)H was eliminated by this mutation, thioesterase activity was completely retained. Using [1-2H]acetaldehyde as a substrate, a significant deuterium isotope effect was observed, implicating Glu377 in the hydride transfer step and not in acylation or release of the acyl group from the cysteine nucleophile. The increase in specificity of the E377Q mutant for NADP+ is consistent with a change in the rate-limiting step determining kcat from nucleotide-dependent NAD(P)H dissociation to hydride transfer. The results provide biochemical evidence that the two highly conserved Glu residues are involved in different functions in the active site of V. harveyi ALDH. PMID- 9219529 TI - Screening mixtures for biological activity by NMR. AB - Development of the new drugs often involves the screening of compound libraries for biological activity. Currently, the biologically active component can only be identified if either a pure compound is being tested or if the components of a mixture are spatially separated, for example, on beads. Here, we present an NMR technique based on the transferred nuclear Overhauser effect (transfer NOE) that allows identification and structural characterization of biologically active molecules from a mixture. As an example we demonstrate that from mixtures of oligosaccharides only alpha-L-Fuc-(1-->6)-beta-D-GlcNAc-OMe binds to Aleuria aurantia agglutinin. The sign of transferred NOEs is opposite to NOEs of small molecules that do not bind to the protein and, thus, an unequivocal identification of molecules with binding activity is possible. Normally, the selection of bound ligands is further facilitated in that the absolute intensity of transfer NOEs is much greater than that of NOEs of non-binding molecules. In addition, transfer NOEs provide information on the three-dimensional structure of the ligands in the bound state. Therefore, measuring transfer NOEs of mixtures of small molecules in the presence of large molecules, like proteins, should significantly enhance the options for screening mixtures of compounds for biological activity. PMID- 9219530 TI - Modulation of the mitochondrial permeability transition by nitric oxide. AB - The influence of nitric oxide on mitochondrial permeability transition (MPT) phenomenon was studied. NO was generated by photolysis of S-nitroso-N acetylcysteine, AcCys(NO), with green light (lambda = 550 nm). Two distinct effects of nitric oxide on rat liver mitochondria were identified. First, NO accelerated an onset of swelling in Ca2(+)-loaded mitochondria in a cyclosporin-A sensitive manner acting as an inducer of permeability transition. This was, apparently, a result of irreversible alteration of mitochondrial function accompanying the inhibition of respiratory chain in the presence of calcium. Formation of ESR-visible iron-sulfur dinitrosyl complexes (g = 2.041) could also contribute to the irreversible changes resulting in MPT induction. Second, NO changed significantly the response of mitochondria to Ca2+/phosphate-induced MPT, acting as a regulator of permeability transition. In this case the action of nitric oxide led to division of the mitochondria into two subpopulations: one which underwent the rapid permeability transition and another in which the MPT was inhibited. The effect of NO on Ca2+/Pi-induced MPT was transient and resulted from reversible inhibition of cytochrome oxidase followed by the changes in transmembrane potential and Ca2+ distribution. The characteristic time of duration of these NO modulated effects depended on nitric oxide as well as on oxygen concentrations. With increasing NO at fixed oxygen concentrations, this time levelled off to reach a maximum value which was inversely related to the oxygen concentration. It is concluded that under physiological condition the duration of reversible NO effects on mitochondrial function could be determined by oxygen concentration. PMID- 9219531 TI - Molecular cloning and expression of a hexamerin cDNA from the malaria mosquito, Anopheles gambiae. AB - During the last larval instar, dipteran insects synthesize two hexamerins rich in aromatic residues, typified by the larval serum proteins 1 and 2 (LSP-1 and LSP 2) of Drosophila melanogaster. We report here the characterization of a complete cDNA sequence encoding a LSP-1-like protein from a lower dipteran insect, the malaria mosquito Anopheles gambiae. The cDNA encodes the subunit of a homohexamer, A. gambiae hexamerin-1.1 (AgHex-1.1), which is a major pupal protein but only a minor constituent of late larval hemolymph. AgHex-1.1 is moderately rich in methionine (3.9%) and particularly rich in aromatic residues (21% Phe+Tyr). Cytogenetic analysis reveals AgHex-1.1 to be encoded by a single-copy gene localized to division 22F within the proximal 2La inversion breakpoint of chromosome 2 of A. gambiae. The AgHex-1.1 transcript is first detected in fourth instar larvae (L4) and disappears abruptly in early pupae. In situ hybridization shows accumulation of the transcript uniquely in the larval fat body. AgHex-1.1 mRNA is re-expressed in male and female adults at about 10% of the L4 level, with no effect of bloodfeeding in females. The potential roles of AgHex-1.1 in Anopheles development and reproductive maturation are discussed. PMID- 9219532 TI - Cloning of the mouse laminin alpha 4 cDNA. Expression in a subset of endothelium. AB - Endothelial cells express a 400-kDa or 240-kDa laminin alpha chain, depending on their tissue of origin or physiological state [1, 2]. Using differential display and subsequent screening of a mouse endothelial cell cDNA library we here identify the gene coding for the 240-kDa laminin chain as the laminin alpha 4 gene. The complete mouse laminin alpha 4 cDNA sequence is reported and compared with other laminin alpha chains. In situ hybridization of embryonic and new born mouse tissues revealed expression of laminin alpha 4 mRNA in a subset of endothelium, in particular aortic endothelium, endocardium and endothelium of blood vessels in the skin and in the brain. Strong laminin alpha 4 expression by aortic endothelia was confirmed by data obtained from cultured bovine aortic endothelial cells (BAEC). Isolation of laminin from BAEC conditioned medium revealed a Y-shaped molecule in rotary shadowing. Subsequent sequencing of BAEC laminin resulted in laminin alpha 4, beta 1 and gamma 1 amino acid sequences, confirming that laminin alpha 4 is one of the major laminin alpha chains expressed by aortic endothelium not only in the mouse. In addition, strong laminin alpha 4 mRNA expression occurred in peripheral nerves, cardiac muscle, fat, the dermis of the skin and lung stroma of mouse tissues. The data demonstrate a cytokine and progesterone-regulated differential expression of laminin alpha 4 mRNA in mouse endothelium, suggesting a distinct functional role for this laminin chain in endothelium. PMID- 9219534 TI - The effect of alpha-tocopherol on the synthesis, phosphorylation and activity of protein kinase C in smooth muscle cells after phorbol 12-myristate 13-acetate down-regulation. AB - Previous work had established that, in smooth muscle cells, alpha-tocopherol negatively regulates protein kinase C by preventing its activation [Tasinato, A., Boscoboinik, D., Bartoli, G. M., Maroni, P. & Azzi, A. (1995) Proc. Natl Acad. Sci. USA 92, 12190-12194]. In this study, the mechanism by which this event takes place has been analyzed. The regulation by alpha-tocopherol of protein kinase C expression, activity and phosphorylation has been followed during the synthesis of protein kinase C after its down-regulation by phorbol 12-myristate 13-acetate. The data show that protein kinase C isoenzyme alpha is synthesised significantly more (30% 72 h after down-regulation) in the presence of alpha-tocopherol. However, its activity is significantly less (45% diminution) and its phosphorylation state is also decreased (60% diminution). The effect of alpha tocopherol appears not to be shared by the analogue beta-tocopherol, provided with similar radical-scavenging properties. The data are interpreted in terms of a diminution of protein kinase C phosphorylation, specifically caused by alpha tocopherol, resulting in a decreased enzyme specific activity. PMID- 9219533 TI - Formation of the early-region-2 transcription-factor-1-retinoblastoma-protein (E2F-1-RB) transrepressor and release of the retinoblastoma protein from nuclear complexes containing cyclin A is induced by interferon alpha in U937V cells but not in interferon-alpha-resistant U937VR cells. AB - We have analysed the different regulation of cell-cycle-relevant proteins by interferon alpha (IFN alpha) in IFN alpha-sensitive and resistant U937 leukemic cell lines. In contrast to the INF alpha-sensitive U937 variant cell line U937V, the IFN alpha-resistant derivative (U937VR) is insensitive to the antiproliferative activity of IFN alpha. As we found no differences between these cell lines concerning the induction by IFN alpha of the pathway involving tyrosine-protein kinases and the signal transducer and activator of transcription (Jak-Stat), we examined whether cell-cycle-regulating proteins are differently affected by IFN alpha in U937VR and U937VR cells. In U937V cells IFN alpha induced the formation of the complex between early-region-2 transcription factor 1 (E2F-1) and retinoblastoma protein (RB) which is known to repress transcription of E2F-1-inducible genes, necessary for cell cycle progression. Formation of this complex was not inducible by IFN alpha in U937VR cells, although the suitable binding partners (E2F-1 and under-phosphorylated RB) were present. Interestingly, treatment of nuclear extracts from logarithmically growing U937V and U937VR cells with an antiserum against cyclin A that disrupts cyclin-A-containing complexes, led to the formation of the E2F-1-RB complex, suggesting the presence of under phosphorylated (active) RB, trapped in nuclear complexes that contain cyclin A. This suggestion was supported by combined immunoprecipitation/western blot experiments that revealed a physical interaction between phosphorylated as well as under-phosphorylated forms of RB and cyclin A complex(es) in U937V and U937VR cells. RB, especially the under-phosphorylated form, was released by treatment with IFN alpha from this complex(es) in the case of U937V cells but not U937VR cells. We conclude that the missing induction of the E2F-1-RB transrepressor by IFN alpha and the failure to release RB from cyclin-A-containing complexes might contribute to the resistance of U937VR cells to the antiproliferative effects of IFN alpha. PMID- 9219535 TI - Structure and expression of the gene encoding secretor-type galactoside 2-alpha-L fucosyltransferase (FUT2). AB - The expression and secretion of ABO antigens in epithelial cells of glands are controlled by secretor-type alpha (1,2)fucosyltransferase activity. We have examined the expression of the secretor-type alpha(1,2)fucosyltransferase gene (FUT2) and a pseudogene of FUT2 (Sec1) in several tumor cell lines by northern blot and/or reverse-transcription-PCR (RT-PCR) analyses. Transcripts of FUT2 were found in total RNA from ovarian, gastric and colonic cancer cell lines but not from six leukemic cell lines, including erythroleukemic HEL cells, by RT-PCR. On the other hand, RT-PCR indicated that Sec1 was expressed in all these tumor cells, including all hematopoietic cells studied. Northern blot analysis indicated that FUT2 transcripts with a similar size (3.3 kb) were expressed in cancer cell lines. Rapid amplification of cDNA ends suggested that the entire FUT2 cDNA is 3.1-kb long and has two Alu repetitive elements in its 3' untranslated region, including an inverted repeat. The mRNA, therefore, may form a large stem-and-loop structure (1.2 kb). Each stem contains about 300 bases, the loop contains 640 bases, and the percentage of complementary nucleotide sequences in the stem region is 85%. The presence of a large stem-and-loop structure in the 3' untranslated region may regulate the level of the FUT2 transcript by affecting the stability of the mRNA. PMID- 9219536 TI - Sequence and expression of an Eisenia-fetida-derived cDNA clone that encodes the 40-kDa fetidin antibacterial protein. AB - Fetidins are 40-kDa and 45-kDa hemolytic and antibacterial glycoproteins present in the coelomic fluid of the earthworm Eisenia fetida andrei. By screening a cDNA library with a polyclonal antifetidin serum, we have cloned a cDNA that encoded the 40-kDa fetidin. The clone contains an insert of 1.44 kb encoding a protein of 34 kDa, which corresponds to the size of deglycosylated fetidins. The recombinant protein inhibits Bacillus megaterium growth. Restriction fragment polymorphisms were observed on Southern blots and correspond to a known protein polymorphism. The sequence of the cDNA contains a peroxidase signature and fetidins from earthworm coelomic fluid have peroxidase activity. The 40-kDa and 45-kDa fetidins therefore represent two related polymorphic defence factors in invertebrates. PMID- 9219537 TI - Two ways of legumin-precursor processing in conifers. Characterization and evolutionary relationships of Metasequoia cDNAs representing two divergent legumin gene subfamilies. AB - Subunit monomers and oligomers of crystalloid-type legumins are major components of SDS-soluble fractions from Metasequoia glyptostroboides (Dawn redwood, Taxodiaceae) seed proteins. The subunits are made up of disulfide linked alpha polypeptides and beta-polypeptides with molecular masses of 33 kDa and 23-25 kDa, respectively. Unusually for legumins, those from Metasequoia are glycosylated and the carbohydrate moieties are residing in the C-terminal region of the respective beta-polypeptides. A Metasequoia endosperm cDNA library has been constructed and legumin-encoding transcripts representing two divergent gene subfamilies have been characterized. Intersubfamily comparisons reveal 75% identity at the amino acid level and the values range from 53-35% when the legumin precursors deduced were compared with those from angiosperms. The predicted sequences together with data from amino acid sequencing prove that post-translational processing of Metasequoia prolegumins is directed to two different processing sites, each of them specific for one of the legumin subfamilies. The sites involved differ in their relative position and in the junction to be cleaved: Metasequoia legumin precursors MgLeg18 and MgLeg26 contain the conventional post-translational Asn Gly processing site, which is generally regarded as highly conserved. In contrast, the MgLeg4 precursor is lacking this site and post-translational cleavage is directed to an unusual Asn-Thr processing site located in its hypervariable region, causing N-terminal extension of the beta-polypeptide relative to those hitherto known. Evidence is given that the unusual variant of processing also occurs in other conifers. Phylogenetic analysis reveals the precursors concerned as representatives of a distinct legumin subfamily, originating from duplication of an ancestral gene prior to or at the beginning of Taxodiaceae diversification. PMID- 9219538 TI - Bov-B long interspersed repeated DNA (LINE) sequences are present in Vipera ammodytes phospholipase A2 genes and in genomes of Viperidae snakes. AB - Ammodytin L is a myotoxic Ser49 phospholipase A2 (PLA2) homologue, which is tissue-specifically expressed in the venom glands of Vipera ammodytes. The complete DNA sequence of the gene and its 5' and 3' flanking regions has been determined. The gene consists of five exons separated by four introns. Comparative analysis of the ammodytin L and ammodytoxin C genes shows that all intron and flanking sequences are considerably more conserved (93-97%) than the mature protein-coding exons. The pattern of nucleotide substitutions in protein coding exons is not random but occurs preferentially on the first and the second positions of codons, which suggests positive Darwinian evolution for a new function. An Ruminantia specific ART-2 retroposon, recently recognised as a 5' truncated Bov-B long interspersed repeated DNA (LINE) sequence, was identified in the fourth intron of both genes. This result suggests that ammodytin L and ammodytoxin C genes are derived by duplication of a common ancestral gene. The phylogenetic distribution of Bov-B LINE among vertebrate classes shows that, besides the Ruminantia, it is limited to Viperidae snakes (Vipera ammodytes, Vipera palaestinae, Echis coloratus, Bothrops alternatus, Trimeresurus flavoviridis and Trimeresurus gramineus). The copy number of the 3' end of Bov-B LINE in the Vipera ammodytes genome is between 62,000 and 75,000. The absence of Bov-B LINE at orthologous positions in other snake PLA2 genes indicates that its retrotransposition in the V. ammodytes PLA2 gene locus has occurred quite recently, about 5 My ago. The amplification of Bov-B LINEs in snakes may have occurred before the divergence of the Viperinae and Crotalinae subfamilies. Due to its wide distribution in Viperidae snakes it may be a valuable phylogenetic marker. The neighbor-joining phylogenetic tree shows two clusters of truncated Bov-B LINE, a Bovidae and a snake cluster, indicating an early horizontal transfer of this transposable element. PMID- 9219539 TI - Solution structure of neuropeptide tyrosine 13-36, a Y2 receptor agonist, as determined by NMR. AB - The three-dimensional structure of neuropeptide tyrosine (NPY) 13-36, a specific Y2 receptor agonist, has been investigated by two-dimensional 1H-NMR spectroscopy in solution. Analysis of the double-quantum-filtered correlation spectroscopy (DQFCOSY), total correlation spectroscopy (TOCSY) and nuclear Overhauser enhancement spectroscopy (NOESY) spectra provided a complete assignment of the proton signals. The interproton connectivities observed in the NOESY spectra comprised 166 intraresidue and 95 interresidue distance ranges which were used as constraints for molecular modeling by distance geometry, simulated annealing and energy minimization. The optimal structures are characterized by a helical C terminal fragment Leu30-Tyr36 and a wide loop from Leu17 to Ser22. The structure of NPY 13-36 is analogous to the structure of NPY under the same solvent conditions. Comparison with other reported Y2 agonists suggests that the helical Leu30-Tyr36 fragment is the most critical for activity. PMID- 9219540 TI - Post-translational processing of rat ribosomal proteins. Ubiquitous methylation of Lys22 within the zinc-finger motif of RL40 (carboxy-terminal extension protein 52) and tissue-specific methylation of Lys4 in RL29. AB - The complete amino acid sequences of rat and yeast (Saccharomyces cerevisiae) ribosomal proteins derived from precursors containing an N-terminal ubiquitin or ubiquitin-like sequence (C-terminal extension proteins or CEPs) were determined and investigated for any post-translational modifications by reverse-phase HPLC purification, direct amino acid sequence and mass spectrometric analyses. Covalent modifications were detected in the rat liver proteins RS27a (CEP-80), RL29, RL37 and RL40 (CEP-52), while RS30 (CEP), RL36a, RL39 and RL41 were unmodified. Heterogeneity of RS27a was due to C-terminal truncations, with Lys80 missing from about 20% of the liver RS27a population; C-terminal processing was also detected with RL29 and RL37. No other covalent modifications of liver, brain or thymus RS27a were detected. The rat RL40 structure was identical to the cDNA predicted sequence except for complete stoichiometric N epsilon-trimethylation of Lys22 within its zinc-finger motif; this modification occurred in the ribosomes of all three rat tissues investigated but not in yeast ribosomes. The methylation characteristics of RL40 were distinct from those of ribosomal protein RL29 in the rat, which was differentially monomethylated at Lys4 in the liver, brain and thymus (27%, > 99% and 95% methylation, respectively). In the case of liver, there was no appreciable difference in the RL29 methylation status of free and membrane-bound ribosomes. The possibilities of an essential role for RL40 methylation in the formation of rat ribosomes, and a distinct regulatory role for RL29 methylation in the rat, are discussed. PMID- 9219541 TI - Translocation to the periplasm and signal sequence cleavage of preapocytochrome c depend on sec and lep, but not on the ccm gene products. AB - Post-translational maturation of soluble cytochrome c includes translocation of the precursor polypeptide and heme through the cytoplasmic membrane, proteolytic cleavage of the signal sequence, and covalent attachment of heme. Specific genes for cytochrome c maturation (ccmABCDEFGH in Escherichia coli) are required for holocytochrome c formation, among them genes encoding an ABC transporter (ccmABC). We investigated the requirements of apocytochrome translocation to the periplasm and characterized specific intermediates of the cytochrome c maturation pathway. Apocytochrome precursor was present in the membrane fraction. Translocation of the polypeptide was independent of ccm gene products, but dependent on a functional secretion machinery, as shown by accumulation of preapocytochrome c in the membranes of secA and secY mutants. After translocation, cleavage of the signal sequence allowed the release of apocytochrome into the periplasm, where heme was bound in a ccm-dependent manner. By contrast, non-cleaved holocytochrome c containing covalently bound heme accumulated in the membranes of a lepB mutant, which indicated that signal sequence cleavage and heme attachment are independent steps in the cytochrome c maturation pathway. PMID- 9219542 TI - Protein and gene structure of the NADH-binding fragment of Rhodobacter capsulatus NADH:ubiquinone oxidoreductase. AB - Membranes of aerobically grown Rhodobacter capsulatus contain only one type of NADH:ubiquinone oxidoreductase which is homologous to the proton-translocating complex I. The K(m) value of the enzyme for NADH was determined to be 8 microM. After solubilization of the membranes with an alkylglucoside detergent, two fragments of complex I with molecular masses of 110 kDa and 140 kDa were isolated by chromatographic steps in the presence of detergent. Both fragments contain at least two polypeptides with apparent molecular masses of 46 kDa and 42 kDa. FMN was identified as cofactor in the preparations. Degenerative oligonucleotide primers were used to amplify a part of the sequence coding for the NADH-binding subunit of complex I by PCR. With the PCR product as probe, a genomic fragment was cloned and sequenced containing the genes encoding the two purified polypeptides and additional reading frames. The two genes are named nuoE and nuoF and are homologous to nqo2 and nqo1 of Paracoccus denitrificans. However, NuoE contains a C-terminal extension of 149 amino acids compared with Nqo2. NuoE and NuoF have molecular masses of 41259 Da and 47133 Da and contain the NADH-, FMN- and FeS-cluster-binding motifs. PMID- 9219543 TI - The three-dimensional solution structure of the lantibiotic murein-biosynthesis inhibitor actagardine determined by NMR. AB - The three-dimensional solution structure of the lantibiotic actagardine was determined at high resolution by homonuclear and heteronuclear two-dimensional and three-dimensional NMR spectroscopy in [2H3]acetonitrile/H2O (7:3). 133 non trivial distance and 22 torsional-angle constraints were derived from the NMR data. An ensemble of 15 low-energy structures was calculated by distance geometry followed by an iterative relaxation-matrix-refinement procedure. The rmsd of the backbone coordinates with respect to the average structure was 17 pm. The two distinct thioether ring systems 1-6 and 7-19 were even better defined, with backbone rmsd of 10 pm and 14 pm, respectively. Actagardine shows a rigid compact globular shape based on the constraining bridging pattern, which is composed of an N-terminal lanthionine ring from residues 1-6 and three intertwined C-terminal methyllanthionine rings comprising residues 7-12, 9-17 and 14-19. In addition, this C-terminal ring system is stabilised by a short antiparallel beta sheet. A feature of the actagardine structure is the presence of two putative binding pockets. A pocket is generated by the covalent constraints of the C-terminal thioether ring system. The rim of this pocket is built up by a loop structure comprising residues 12-19, whose backbone amide protons are all directed to the centre of the pocket. The second pocket is formed by an L-shaped orientation of the N-terminal and C-terminal thioether ring systems. The only two hydrophilic amino acid residues of actagardine, Glu11 and Ser2, are directed to this pocket. A region of high sequence similarity with the related lantibiotic mersacidin is located exactly at the position of the second pocket (residues 3-12). This suggests that the second pocket is responsible for the antibiotic mode of action of actagardine and mersacidin as inhibitors of the murein biosynthesis of gram positive bacteria. PMID- 9219544 TI - Envelope protein of parasitic wasp symbiont virus, polydnavirus, protects the wasp eggs from cellular immune reactions by the host insect. AB - Cotesia kariyai polydnavirus (CkPDV) virions are present in the oviducts of C. kariyai wasp and are injected with eggs into the hemocoel of the host armyworm Pseudaletia separata larvae during parasitization. Evidence that the presence of polydnavirus particles on the surface of the wasp eggs may be essential for prevention of cellular immune reactions by the host hemocytes was obtained by isolating an immunoevasive factor from CkPDV virions. The purified proteinaceous factor protects foreign materials from adhesion and encapsulation by hemocytes of the host P. separata larvae but not by those of common cutworm Spodoptera litura larvae which is an incompatible host for the C. kariyai wasp. Purification procedures consisted of extraction with ethanol/trifluoroacetic acid and reverse phase high performance liquid chromatography. A factor with a molecular mass of approximately 50 kDa is demonstrated to be present on the envelope of CkPDV virion by immunoelectronmicroscopic observations. Furthermore, immunoreactive proteins are found in plasma of the armyworm larvae but not in the common cutworm larvae, indicating that only the natural host of C. kariyai wasp shares a similar epitope with CkPDV. The sequence of 23 amino acid residues at the amino terminus of the factor was determined to be Ile-Ser-Val-Glu-Asn-Val-Xaa-Thr-Thr-Gly-Ile Phe-Leu-Asp-Ser-Gly-Glu-Xaa- Val- Pro-Tyr-Ala-Thr-Lys-Pro. PMID- 9219545 TI - Methods for predicting carcinogenic hazards: new opportunities coming from recent developments in molecular oncology and SAR studies. AB - Without epidemiological evidence, and prior to either short-term tests of genotoxicity or long-term tests of carcinogenicity in rodents, an initial level of information about the carcinogenic hazard of a chemical that perhaps has been designed on paper, but never synthesized, can be provided by structure-activity relationship (SAR) studies. Herein, we have reviewed the interesting strategies developed by human experts and/or computerized approaches for the identification of structural alerts that can denote the possible presence of a carcinogenic hazard in a novel molecule. At a higher level of information, immediately below epidemiological evidence, we have discussed carcinogenicity experiments performed in new types of genetically engineered small rodents. If a dominant oncogene is already mutated, or if an allele of a recessive oncogene is inactivated, we have a model animal with (n-1) stages in the process of carcinogenesis. Both genotoxic and receptor-mediated carcinogens can induce cancers in 20-40% of the time required for classical murine strains. We have described the first interesting results obtained using these new artificial animal models for carcinogenicity studies. We have also briefly discussed other types of engineered mice (lac operon transgenic mice) that are especially suitable for detecting mutagenic effects in a broad spectrum of organs and tissues and that can help to establish mechanistic correlations between mutations and cancer frequencies in specific target organs. Finally, we have reviewed two complementary methods that, while obviously also feasible in rodents, are especially suitable for biomonitoring studies. We have illustrated some of the advantages and drawbacks related to the detection of DNA adducts in target and surrogate tissues using the 32P-DNA postlabeling technique, and we have discussed the possibility of biomonitoring mutations in different human target organs using a molecular technique that combines the activity of restriction enzymes with polymerase chain reaction (RFLP/PCR). Prediction of carcinogenic hazard and biomonitoring are very wide ranging areas of investigation. We have therefore selected five different subfields for which we felt that interesting innovations have been introduced in the last few years. We have made no attempt to systematically cover the entire area: such an endeavor would have produced a book instead of a review article. PMID- 9219546 TI - Comparison of the lethal effects of different actinomycins on a repair-deficient strain of Escherichia coli. AB - We report here the isolation and purification of a genotoxic antibiotic from the culture medium of a Streptomyces strain. The antibiotic was identified as actinomycin X2 by using a database search (AntiBase). In the previous studies a related compound, actinomycin D, has been shown to be non-mutagenic in the salmonella/microsome assay and several other bacterial systems. The fact that an actinomycin was detected by a bacterial repair assay seems to contradict these results. Therefore we tested several actinomycins by differential killing assay based on the same Escherichia coli strains that were used in our screening bioassays. According to our results the uvrA, recA double mutant sensitizes E. coli to the genotoxic effects of actinomycins. PMID- 9219548 TI - An improved method for the mouse liver micronucleus test. AB - We have developed a very practical method for performing the liver micronucleus test in mice. Using this method, we evaluated 11 different types of mutagens, including, 2-acetylaminofluorene, amsacrine, benzene, cyclophosphamide, diethylnitrosamine, 4-dimethylamino-3'-methylazobenzene, N-ethyl-N-nitrosourea, fluorouracil, mitomycin C, potassium chromate (VI) and selenious acid. In order to assess the sensitivity of our method, the peripheral blood reticulocyte micronucleus test was performed in the same mouse. Animals were given test chemicals once and underwent partial hepatectomy (PH) 24 h later in order to induce mitotic stimulation. Peripheral blood was sampled 0, 24, 48 and 72 h after treatment. The incidence of micronucleated hepatocytes was determined 5 days after PH. As a result, diethylnitrosamine and 4-dimethylamino-3' methylazobenzene, known as liver carcinogens, increased the incidence of micronucleated cells in the liver only. Positive reactions for benzene, on the other hand, were found in the peripheral blood reticulocytes only. The other chemicals showed positive reaction in the liver and peripheral blood reticulocytes with almost the same maximum response of micronucleus induction. Our method was found to have the advantage over Cliets' liver micronucleus test in that it required much less time and was easier to perform procedures and highly sensitive in detecting clastogens. It can be used in combination with the peripheral blood reticulocyte micronucleus test to evaluate test chemicals in two tissues, the liver and the bone marrow, in the same animal. We propose a method of combining this test with the peripheral blood reticulocyte micronucleus test for efficient screening for the clastogenic potential of new chemicals in vivo. PMID- 9219547 TI - Genotoxic effects of three Fusarium mycotoxins, fumonisin B1, moniliformin and vomitoxin in bacteria and in primary cultures of rat hepatocytes. AB - The genotoxic effects of three widespread Fusarium toxins, vomitoxin (VOM), moniliformin (MON) and fumonisin B1 (FB1) were investigated in bacterial tests and in micronucleus (MN) and chromosomal aberration (CA) assays with primary rat hepatocytes. All three toxins were devoid of activity in gene mutation assays with Salmonella typhimurium strains TA98 and TA100 and in SOS chromotests with E. coli strain PQ37 in the presence and absence of metabolic activation. FB1 and VOM gave negative results in differential DNA repair assays with E. coli K-12 strains (343/753, uvrB/recA and 343/765, uvr+/rec+); with MON, a marginal effect was seen in the absence of metabolic activation mix at relatively high concentrations (> or = 55 micrograms/ml). In metabolically competent rat hepatocytes stimulated to proliferate with EGF and subphysiological Ca2+ concentrations, a decrease of cell division was observed with all three toxins at concentrations > or = 10 micrograms/ml, VOM was strongly cytotoxic at 100 micrograms/ml. All three mycotoxins caused moderate increases of the MN frequencies at low concentrations (< or = 1 microgram/ml), but no clear dose-response effects were seen and at higher exposure levels the MN frequencies declined. In the CA experiments with hepatocytes, pronounced dose-dependent effects were observed with all three toxins. MON caused a 9-fold increase over the spontaneous background level after exposure of the cells to 1 microgram/ml for 3 h, with FB1 and VOM, the increases were 6- to 7-fold under identical experimental conditions. This is the first report on clastogenic effects of VOM and FB1 in mammalian cells, with MON induction of CAs in V-79 cells has been described earlier. Since all three mycotoxins caused CAs at very low concentration levels in liver cells in vitro, it is possible that such effects may also occur in humans and mammals upon consumption of Fusarium-infected cereals. PMID- 9219549 TI - Dairy cattle as a bioindicator of exposure to genotoxic substances in a heavily polluted area in northern Bohemia. AB - The exposure of dairy cattle to genotoxic agents in two districts with different levels of environmental pollution was estimated using cytogenetic analysis of bovine peripheral lymphocytes. The Teplice district represented an industrialized area where the air pollution rate is extremely high mainly in the winter, and the Prachatice district--an agricultural area with a relatively low level of pollution. The Ames test was used to examine feed samples for the content of mutagenic substances. Cows in the Teplice district showed a significantly higher count of aberrant cells (4.83 +/- 2.36) than cows in the Prachatice district (3.63 +/- 2.12). The sum of revertants induced by rinsings or extracts of feeds in both of the two test strains (Salmonella typhimurium TA 98 and TA 100) was significantly higher in the district of Teplice than in the district of Prachatice. The percentages of findings with mutagenic responses were 56.3 and 34.8% for the districts of Teplice and Prachatice, respectively. No mutagenic activity was found in milk samples collected in any of the districts. Apparently, the cows kept in the Teplice district were more exposed to genotoxic substances than the cows in the Prachatice district. The major source of this exposure was probably fresh fodder contaminated by industrial emissions. PMID- 9219550 TI - Exposure of organic extracts of air particulates to sunlight leads to metabolic activation independence for mutagenicity. AB - Air particulates were collected on Whatman, GFA glass fibre filters using a RADECO constant-flow air sampler from a car-parking basement and an open roadside adjacent to the basement. While the basement was not exposed to sunlight, the roadside from where air samples were collected was exposed to regular daylight in the month of July (peak summer month). The filters were soaked and sonicated in acetone to dislodge the particulates and then a residue was obtained after evaporation of acetone. The residues were either held in dark or exposed to natural sunlight or germicidal UV light before being tested for mutagenicity using the Salmonella tester strain TA98 with and without metabolic activation (S9 mix). The results showed that the addition of S9 mix resulted in only a slight increase in the frequency of histidine revertants/plate in the case of daylight exposed roadside air samples. On the other hand, a considerable increase in mutagenicity was observed in the case of the basement air samples, particularly at higher concentrations of the organic extracts when S9 mix was added. However, a pre-exposure of the organic extract of air from the basement to sunlight abrogated the need for S9 mix for showing mutagenic activity. A pre-exposure of the same extracts to germicidal UV light failed to produce a similar effect. These results suggested that long wavelengths of natural sunlight could be responsible for the conversion of certain promutagens in air particulates into direct-acting mutagens. The environmental impact of solar radiation as a modifier of air particulate mutagens in high-sun countries like Saudi Arabia needs to be carefully considered for assessment of air pollution-related health risks. PMID- 9219551 TI - Preferential formation and decreased removal of cisplatin-DNA adducts in Chinese hamster ovary cell mitochondrial DNA as compared to nuclear DNA. AB - Levels of DNA adducts in Chinese hamster ovary (CHO) cells exposed to cis diamminedichloroplatinum(II) (cisplatin) for 24 h, have been shown to be 4- to 6 fold higher in mitochondrial (mt) DNA as compared to nuclear (n) DNA (Olivero et al., Mutation Res., 346 (1995) 221). The aim of the present study was to understand if the preferential cisplatin binding in mtDNA is partially caused by lack of adduct removal in the mitochondria. Chinese hamster ovary cells were exposed for 6 h to 50 microM cisplatin, followed by incubation for 24 and 48 h in cisplatin-free medium. At the 30-h time point (6 h with cisplatin, 24 h without cisplatin), half of the cells from each plate were harvested and the remainder were cultured and harvested at 54 h (6 h with cisplatin, 48 h without cisplatin). The 30- and 54-h time points are called 'T30' and 'T54', respectively. Cisplatin DNA adducts were measured in DNA from nuclear and mitochondrial fractions by dissociation-enhanced lanthanide fluoroimmunoassay (DELFIA), a sensitive competitive microtiter-based immunoassay utilizing antiserum elicited against cisplatin-modified DNA. An initial higher level of cisplatin-DNA adducts was observed in mtDNA when compared to nDNA, at T30. In addition, a lack of removal of adducts in mtDNA was demonstrated in cells at T54. Dilution of DNA adducts by DNA replication was documented in pulse-chase experiments that employed [3H]thymidine incorporation. Adduct removal by repair-related mechanisms was considered to comprise the difference between total DNA adduct removal and adduct removal related to DNA replication. The final results demonstrated that both, higher initial binding and lack of removal of cisplatin-DNA adducts appear to contribute to the preferential cisplatin-mtDNA binding observed in CHO cells. PMID- 9219552 TI - A direct comparison of mouse and rat bone marrow and blood as target tissues in the micronucleus assay. AB - Rats and mice were treated concurrently with mitomycin C at a dose of 1 mg/kg/day i.p. for 3 days, a regimen known to induce micronuclei in polychromatic erythrocytes (MN-PCE) in the bone marrow of rats and mice and the peripheral blood of mice. The incidence of micronuclei was evaluated in the peripheral blood and the bone marrow of both species. Early reports suggested that the efficiency of the rat spleen in removing micronuclei from the circulation precluded the use of rat peripheral blood in the detection of chemically-induced micronuclei. The data in the present study demonstrate that the induction of micronuclei in polychromatic erythrocytes as the result of treatment with a clastogen can be demonstrated equally well in the bone marrow or the peripheral blood of both rats and mice. PMID- 9219553 TI - A comparative study of synchronised and conventional culture methods on the micronucleus dose-response curve. AB - Micronucleus (MN) dose-response curves have been studied in blood samples obtained from a healthy volunteer with both a methotrexate synchronised culture and with a conventional culture method. The curve obtained with the synchronised culture, showed better response at higher doses compared to that obtained with the conventional procedure. Generally, MN frequency obtained at a dose of 4.0 Gy with the conventional procedure is lower, compared to that obtained with dicentric (DC) frequency. The present study also showed that MN frequency obtained at this dose was lower (0.79 +/- 0.09) compared to that obtained with DC frequency (0.91 +/- 0.10). However, DC frequency obtained with the synchronised culture was almost the same as that obtained with the conventional method, whereas acentric frequency showed an increase with the synchronised culture. The study showed that the ratios of MN frequency/total aberration frequency observed with the synchronised (0.63) and that obtained with the conventional culture (0.59) was more or less the same indicating that higher acentric frequency may be the cause for higher MN frequency in the synchronised culture. The present study indicates that methotrexate may not be the cause of higher acentric frequency in synchronised culture. A possible reason for the higher acentric frequency in the synchronised culture, is highlighted. PMID- 9219554 TI - The use of a urine mutagenicity assay in the monitoring of environmental exposure to genotoxins. AB - Urinary bacterial mutagenicity was used as a biomarker of exposure to ambient air pollution in a group of women working outdoors in the city of Teplice (TP; Northern Bohemia) with higher levels of air pollution than a similar group of women in the city of Prachatice (PT; Southern Bohemia). The Salmonella typhimurium plate incorporation assay with the TA98 and YG1041 strains and microsuspension assay with the YG1041 strain were used for testing the urinary mutagenicity. PAH and their metabolites were analyzed by HPLC and GC/MS methods. The significantly higher values of most PAHs/metabolites detected in a TP group confirmed the differences of PAH exposures between both groups. In the plate incorporation assay, the TA98 strain was not able to detect the increase in urinary mutagenicity, but, for the YG1041 strain, the urinary mutagenicity was clearly determined with a significant difference in number of YG1041 + S9 revertants between the TP and PT groups. The microsuspension assay increased the mean response by about 10-fold over the standard plate test; however, no statistical difference between TP and PT groups was found due to high interindividual variability and small sample size. Comparing the urinary PAH/metabolites to urinary mutagenicity, significant correlations were observed between the plate incorporation mutagenicity results with the YG1041 revertants in the presence of metabolic activation and several of the urinary PAH/metabolites. On the contrary, in the microsuspension assay, several urinary PAH/metabolites correlated significantly with the YG1041 revertants only in the absence of metabolic activation. This may indicate the influence of different treatment conditions of assays on the urinary mutagenicity results. The results suggest the insufficient sensitivity of the TA98 tester strain to determinate low urinary level of mutagens. On the contrary, the use of the YG1041 tester strain increases the probability of detecting an effect of environmental exposure and seems to be applicable to biological monitoring. To definitely replace the standard plate incorporation assay with the microsuspension method is not possible without further comparative studies. PMID- 9219555 TI - Studies on the relationship between treatment condition and micronucleus induction in V79 cells exposed to silica and glass fibers. AB - Studies have been carried out to determine the relationship between treatment condition and frequencies of micronucleated cells (MNC) and multinucleated cells (MTC) in Chinese hamster lung fibroblasts (V79 cells) exposed to dusts and fibers. Cells were treated with Min-U-Sil 5 silica or Owens Corning AAA-10 glass fibers under three different conditions: 24-h exposure (24E), 24-h exposure followed by 24-h post-incubation in fresh medium (24E-24P), and 48-h exposure (48E). Results showed that the frequency of MNC increased in a concentration related manner in silica-treated V79 cells only under the condition of 24E-24P. The increase in MNC frequency after 24-h exposure was not concentration-related. No significant increase in MNC was detected in cells sampled after 48-h treatment. The frequencies of MTC in the treatment groups were higher than that in the control group. However, the increase was not statistically significant. Compared with silica, glass fibers were more active for MTC and MNC induction on a mass basis. The highest response was also observed under the condition of 24E 24P. These results indicate that 24-h exposure followed by 24-h post-incubation is a suitable treatment condition for the micronucleus assay on mineral dusts and fibers. PMID- 9219556 TI - Arsenic: a paradoxical human carcinogen. PMID- 9219557 TI - Relationship of urinary arsenic to intake estimates and a biomarker of effect, bladder cell micronuclei. AB - The purpose of this study was to investigate methods for ascertaining arsenic exposure for use in biomarker studies. Urinary arsenic concentration is considered a good measure of recent arsenic exposure and is commonly used to monitor exposure in environmental and occupational settings. However, measurements reflect exposure only in the last few days. To cover longer time periods exposure can be estimated using arsenic intake data, calculated by combining measures of environmental arsenic and inhalation/ingestion rates. We compared these different exposure assessment approaches in a population chronically exposed to arsenic in drinking water in northern Chile. The study group consisted of 232 people, some drinking water low in arsenic (15 micrograms/l) and others drinking water with high arsenic concentrations (up to 670 micrograms/l). First morning urine samples and questionnaire data, including fluid intake information, were collected from all participants. Exfoliated bladder cells were collected from male participants for the bladder cell micronuclei assay. Eight different indices of exposure were generated, six based on urinary arsenic (microgram As/l urine; microgram As/g creatinine; microgram InAs/l urine; microgram MMA/l urine; microgram DMA/l urine; microgram As/h, excreted), and two on fluid intake data (microgram As/day, ingested; microgram As/l fluid ingested-day). The relationship between the different exposure indices was explored using correlation analysis. In men, exposure indices were also related to a biomarker of effect, bladder cell micronuclei. While creatinine adjusted urinary arsenic concentrations had the strongest correlations with the two intake estimates (r = 0.76, r = 0.81), unadjusted urinary arsenic showed the strongest relationship with bladder cell micronuclei. These data suggest that, in the case of the bladder, unadjusted urinary arsenic concentrations better reflect the effective target organ dose compared to other exposure measures for biomarker studies. PMID- 9219558 TI - Arsenic methylation capacity, body retention, and null genotypes of glutathione S transferase M1 and T1 among current arsenic-exposed residents in Taiwan. AB - In order to elucidate the relationships among arsenic methylation capacity, body retention, and genetic polymorphisms of glutathione S-transferase (GST) M1 and T1, a total of 115 study subjects were recruited from Lanyang Basin located on the northeast coast of Taiwan. Specimens of drinking water, blood, urine, hair and toenail were collected from each study subject. Urinary inorganic and methylated arsenic were speciated by high performance liquid chromatography combined with hydride-generation atomic absorption spectrometry. Arsenic concentration in hair and toenail were quantitated by atomic absorption spectrophotometry. The polymerase chain reaction was used to determine genetic polymorphisms of GST M1 and T1. Arsenic concentrations in urine, hair, and toenail of study subjects were positively correlated with arsenic levels in their drinking water. Percentages of various arsenic species in urine (mean +/- standard error (SE) were 11.8 +/- 1.0, 26.9 +/- 1.2 and 61.3 +/- 1.4, respectively, for inorganic arsenic, monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA). Men and women had similar arsenic methylation capability. No associations were observed between arsenic methylation capability and arsenic content in either drinking water or urine. Ratios of arsenic contents in hair and toenail to urinary arsenic content (mean +/- standard error) were 6.2 +/- 0.7 and 16.5 +/- 1.7, respectively. Genetic polymorphisms of GST M1 and T1 were significantly associated with arsenic methylation. Subjects having the null genotype of GST M1 had an increased percentage of inorganic arsenic in urine, while those with null genotype of GST T1 had an elevated percentage of DMA in urine. Arsenic contents in hair and toenail were significantly correlated with the increase in arsenic concentrations of drinking water and urine, while no significant associations were observed between arsenic contents in hair and toenail and polymorphisms of GST M1 and T1. The relationship between arsenic methylation capability and body retention was modified by genetic polymorphisms of GST M1 and T1. Arsenic contents in hair and toenail were negatively associated with MMA percentage and positively associated with DMA percentage among subjects having null genotypes of GST M1 and T1, but not among those with non-null genotypes. PMID- 9219559 TI - Arsenic can mediate skin neoplasia by chronic stimulation of keratinocyte-derived growth factors. AB - Although numerous epidemiological studies have shown that inorganic arsenicals are human skin carcinogens, there is currently no accepted mechanism for its action or an established animal model for its study. We observed increased mRNA transcripts and secretion of keratinocyte growth factors, including granulocyte macrophage-colony stimulating factor (GM-CSF) and transforming growth factor alpha (TGF-alpha) and the proinflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) in primary human epidermal keratinocytes cultured in the presence of low micromolar concentrations of sodium arsenite. Total cell numbers, as well as c-myc expression and incorporation of [3H]thymidine, both indicators of cell proliferation, were also elevated in keratinocyte cultures treated with sodium arsenite. As an in vivo model, the influence of arsenic on mouse skin tumor development was studied in transgenic TG.AC mice which carry the v-Ha-ras oncogene, and can serve as a genetically initiated model for skin carcinogenesis. Following low-dose application of 12-O-tetradecanoyl phorbol-13-acetate (TPA), a marked increase in the number of skin papillomas occurred in transgenic mice receiving arsenic in the drinking water as compared to control drinking water. Papillomas did not develop in arsenic-treated transgenic mice that had not received TPA or arsenic-treated wild-type FVB/N mice, suggesting that arsenic is neither a tumor initiator or promoter but rather an enhancer. Injection of anti GM-CSF antibodies following application of TPA in transgenic mice reduced the number of papillomas. Consistent with that observed in human keratinocyte cultures, increases in GM-CSF and TGF-alpha mRNA transcripts were found within the epidermis of arsenic-treated mice when compared to controls within 6 weeks of treatment. These results suggest that arsenic enhances papilloma development via the chronic stimulation of keratinocyte-derived growth factors and represents the first example of a chemical carcinogen that acts in this manner. These studies suggest that in vitro studies with human keratinocyte cultures examined in conjunction with TG.AC transgenic mice can provide a useful model for examining the tumor enhancing properties of environmental chemicals. PMID- 9219560 TI - Cytogenetic effects in human exposure to arsenic. AB - The cytogenetic effects of arsenic exposure were studied among rural populations that live in the same geographical area and have similar socioeconomic status, but different degree of exposure to inorganic arsenic (As) via drinking water. A group of inhabitants of Santa Ana (408.17 micrograms/l of As in drinking water) were considered the exposed individuals and a group of inhabitants of Nazareno (29.88 micrograms/l) were considered as controls. Blood and urine samples were obtained from volunteers. Past and current exposure, health, and nutritional status as well as the presence of arsenic skin lesions were ascertained in study participants through questionnaires and physical examination. The frequencies and types of chromosomal aberrations in first-division metaphases were studied in whole blood lymphocyte cultures while the presence of micronuclei (MN) was studied in exfoliated epithelial cells obtained from the oral mucosa and from urine samples. Total arsenic (TAs) content, and the relative proportions of inorganic arsenic (IAs), and the metabolites monomethylarsonic (MMA) and dimethylarsinic (DMA) acid were determined in urine samples. Exposed individuals showed a significant increase in the frequency of chromatid and isochromatid deletions in lymphocytes and of MN in oral and urinary epithelial cells. Males were more affected than females, and a higher number of micronucleated oral cells were found among those individuals with skin lesions. The type of cytogenetic damage observed gives evidence of arsenic as a clastogenic/aneugenic carcinogen. PMID- 9219561 TI - Enzymatic methylation of arsenic compounds: IV. In vitro and in vivo deficiency of the methylation of arsenite and monomethylarsonic acid in the guinea pig. AB - Using an in vitro assay which measures the transfer of a radiolabeled methyl moiety of S-[methyl-3H]adenosylmethionine ([3H]SAM) to arsenite or monomethylarsonate (MMA) to yield [methyl-3H]MMA or [methyl-3H]dimethylarsinate (DMA) respectively, guinea pig liver cytosol was found to be deficient in the enzyme activities which methylate these substrates. Moreover, when guinea pigs were given a single intraperitoneal dose of [73As]arsenate (400 micrograms/kg body weight, 25 microCi/kg body weight), very little or no methylated arsenic species were detected in the urine after cation exchange chromatography. The urine collected 0-12 h after arsenate injection contained 98% inorganic arsenic and less than 1% DMA. No MMA was detected in the 0-12 h urine. Urine collected 12 24 h after injection contained approximately 93% inorganic arsenic, 2% MMA and 3% DMA in five of the six animals studied. However, in the 12-24 h urine of one guinea pig, 17% of the radioactivity was DMA, 80% was inorganic arsenic and 3% was MMA. The guinea pig, like the marmoset and tamarin monkeys and unlike most other animals studied thus far, appears to be deficient as far as the enzyme activities that methylate inorganic arsenite. The results of these experiments suggest that there may be a genetic polymorphism associated with the enzymes that methylate inorganic arsenite. PMID- 9219562 TI - Spontaneous and induced sister chromatid exchanges and delayed cell proliferation in peripheral lymphocytes of Bowen's disease patients and matched controls of arseniasis-hyperendemic villages in Taiwan. AB - A total of 15 newly-developed Bowen's disease patients and 34 age-sex-residence matched controls were recruited from three arseniasis-hyperendemic villages in Taiwan to compare spontaneous and arsenic-induced sister chromatid exchanges (SCEs), proportion of cells with high frequencies of SCEs (HFCs), and replication index (RI) in their peripheral lymphocytes. Arsenic-induced Bowen's disease patients were found to have significantly higher spontaneous SCEs and HFCs and a lower spontaneous RI than in matched controls without or with adjustment for age, gender, cigarette smoking, alcohol drinking, tea drinking, status of major diseases, HBsAg carrier status and arsenic exposure indices through multivariate analysis. Sodium arsenite was found to increase SCEs and HFCs and to decrease RI in a dose-response pattern for both cases and controls. The arsenic-induced decrease in RI was significantly greater in arsenic-induced Bowen's disease patients than in matched controls. The arsenic-induced increases in SCEs and HFCs were also consistently, but not statistically significantly, higher in arsenic induced Bowen's disease patients than in matched controls at all arsenite treatment levels of 0.5, 1.0 and 2.0 microM. The arsenic-induced increase in cytogenetic damages and decrease in cell proliferation among arsenic-induced Bowen's disease patients compared with matched controls may result from their long-term exposure to inorganic arsenic through consumption of high-arsenic artesian well water, elevated individual genetic and acquired susceptibility to arsenic-induced damage, or both. PMID- 9219563 TI - Arsenate suppression of human keratinocyte programming. AB - The human keratinocyte line SCC-9 has been used as a model for arsenate-induced perturbations of differentiation. Growth of these cells in 10 microM arsenate permitted the cultures to reach confluence, but prevented expression of 6 markers of suprabasal differentiation (involucrin, loricrin, filaggrin, spr 1, keratin 1 and keratin 10) as assayed by Northern blotting. By contrast, only slight alterations in mRNA levels were observed for one differentiation marker (keratinocyte transglutaminase) and for keratin 5, keratin 14, AP2 or glyceraldehyde phosphate dehydrogenase. The transition metal oxyanions vanadate and chromate had essentially the same suppressive effect on these markers as arsenate, while chronic treatment with tetradecanoylphorbol acetate was generally less effective in suppressing differentiation. To determine whether the previously observed arsenate-mediated alteration in AP1 and AP2 activities could account for the suppression of involucrin, a promoter analysis was conducted. Putative AP1 and AP2 response elements were identified in regions important for transcriptional activity of the 5'-flanking DNA. Mutations in two AP1 sites and one AP2 site were observed to decrease promoter activity significantly, and in combination, to reduce it to approximately 10% of that conferred by the native sequence. These results lend support to the working hypothesis that arsenate suppresses involucrin expression, and, more generally, keratinocyte programming, by altering the transcription factors AP1 and AP2. PMID- 9219564 TI - Arsenic alters cytosine methylation patterns of the promoter of the tumor suppressor gene p53 in human lung cells: a model for a mechanism of carcinogenesis. AB - Arsenic is a potent human carcinogen to which there is significant worldwide exposure through natural contamination of food and drinking water sources. Because arsenic is detoxified via methylation using a methyltransferase (MTase) and S-adenosylmethionine (SAM) as the methyl donor, we hypothesized that a mechanism of carcinogenesis of arsenic could involve alterations of MTase/SAM dependent DNA methylation of a tumor suppressor gene. We found that exposure of human lung adenocarcinoma A549 cells to sodium arsenite (0.08-2 microM) or sodium arsenate (30-300 microM), but not dimethylarsenic acid (2-2000 microM), produced significant dose-responsive hypermethylation within a 341-base pair fragment of the promoter of p53. This was determined by quantitative PCR/HpaII restriction site analysis to analyze methylation status of two CCGG sites. In experiments with arsenite, DNA sequencing using bisulfite to visualize 5-methylcytosine (5 MeC) over the entire promoter region confirmed data obtained by restriction analysis. Limited data using SssI methylase also suggested that over-methylation of CpG sequences may exist over the entire genome in response to arsenite exposure. We propose that alteration of DNA methylation by arsenic offers a plausible, unified hypothesis for the carcinogenic mechanism of action of arsenic, and we present a model for arsenic carcinogenesis that utilizes perturbations of DNA methylation as the basis for the carcinogenic effects of arsenic. PMID- 9219565 TI - Relative genotoxic potency of arsenic and its methylated metabolites. AB - Arsenic is one of the few identified human carcinogens that has yet to be shown to cause cancer in rodents when the standard bioassay protocols are used. The reasons for this apparent interspecies difference are unclear but may be related to differences between humans and rodents in their detoxification capabilities. Detoxification of arsenic may occur through a methylation pathway. If, in fact, methylation does detoxify arsenic, one would predict that the methylated arsenicals might be less genotoxic than the inorganic arsenicals. To evaluate the hypothesis that the inorganic arsenicals are more mutagenic than the organic arsenicals, we tested sodium arsenite, sodium arsenate, monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA) for their relative mutagenic and clastogenic potentials. We used the L5178Y/TK+/- mouse lymphoma assay which allows the detection of chemicals inducing a broad spectrum of different types of genetic damage. Sodium arsenite and sodium arsenate were active at concentrations of 1-2 micrograms/ml and 10-14 micrograms/ml, respectively. MMA was active between 2500 5000 micrograms/ml; while DMA required almost 10000 micrograms/ml to induce a genotoxic response. The organic arsenicals are thus much less potent as mutagenic agents than the inorganic arsenicals. All four of these arsenicals appear to act by mechanisms that cause chromosomal mutations. PMID- 9219567 TI - Variation in arsenic-induced sister chromatid exchange in human lymphocytes and lymphoblastoid cell lines. AB - This study was undertaken to compare the genotoxic effects of arsenite in cultured human lymphocytes and lymphoblastoid cell lines from a group of normal human volunteers. The goal was to determine whether, as found with other genotoxins, subgroups might exist which showed relative high or low sensitivity to induction of sister chromatid exchanges (SCEs) by this metal. Primary lymphoblast cultures were established by treatment with phytohemagglutinin (PHA L). Lymphoblastoid cell lines were established by transformation with Epstein Barr virus. Cultures were exposed for 40 h to sodium arsenite (AsIII) and SCEs assayed by 5-bromo-2'-deoxyuridine incorporation and staining by fluorescence plus Giemsa. SCEs were increased by arsenite in a dose-dependent manner over the concentration range of 10(-7)-10(-5) M. SCEs could not be scored above 10(-5) M because of cytotoxicity. Comparison of SCE frequency in primary lymphocyte cultures among individuals showed substantial variation in sensitivity to arsenite, with some showing no significant effect while others showed a 2-3-fold increase in SCE frequency. In one lymphoblastoid cell line especially sensitive to arsenite, arsenic acid (AsV) or dimethylarsinic acid (DMA) at concentrations up to 10(-5) M did not increase the SCE frequency suggesting that AsIII is the active form of arsenic. When pooled data from the primary lymphocytes was compared to that obtained with the lymphoblastoid cells, the slopes of the dose response curves for ASIII-induced SCEs were similar. The sensitivity of the majority of the individual primary lymphocyte cultures to SCE induction by arsenite was correlated with the sensitivity of the lymphoblastoid cultures established from the same individual. However, in three individuals no correlation was found. Individual lymphoblastoid cell lines retained their As sensitivity after cryopreservation and subsequent revival. Whether the genotoxic response to As is genetically controlled or the result of phenotypic selection is being explored in these stable lymphoblastoid cell lines. PMID- 9219566 TI - Disruption of microtubule assembly and spindle formation as a mechanism for the induction of aneuploid cells by sodium arsenite and vanadium pentoxide. AB - Arsenic and vanadium are important environmental and industrial pollutants. Due to their widespread occurrence and potential genotoxicity, we studied the aneuploidy-inducing effects of these elements in cultured human lymphocytes using a variety of techniques including fluorescence in situ hybridization (FISH) with DNA probes for chromosomes 1 and 7, immunostaining of the lymphocyte spindle apparatus, and an in vitro assay measuring the polymerization and depolymerization of tubulin. Dose-related increases in hyperdiploidy were seen in lymphocyte cultures treated with sodium arsenite (NaAsO2) or vanadium pentoxide (V2O5) over concentrations ranging from 0.001 to 0.1 microM. NaAsO2-treated cells from different donors exhibited similar hyperdiploid frequencies, whereas substantial inter-individual variability was seen in the V2O5-treated cells. Examination of the spindle apparatus using an anti-beta-tubulin antibody indicated that these compounds might disrupt spindle formation by interacting with microtubules. Additional in vitro assays using purified tubulin indicated that both compounds inhibited microtubule assembly and induced tubulin depolymerization. These results indicate that in vitro exposure to both NaAsO2 and V2O5 can induce aneuploidy in human lymphocytes, and that this effect may occur through a disruption of microtubule function. PMID- 9219568 TI - Human cells lack the inducible tolerance to arsenite seen in hamster cells. AB - Chinese hamster V79 cells and their arsenite-resistant variants were found to have an arsenite- and antimonite-inducible tolerance mechanism which protects against the subsequent cytotoxic effects of arsenate, arsenate and antimonite. Inducible tolerance requires de novo mRNA and protein synthesis, and is independent of the heat shock response. In contrast, we report that the arsenite hypersensitive variant line As/S27D lacks the inducible tolerance response. Numerous attempts were made to detect an inducible tolerance response to arsenite in a variety of human cells. An assay based on Neutral red uptake was used in order to study inducible tolerance in cells with poor clonability. Neither normal diploid cells nor human tumor cells of different origins were found to elicit an inducible tolerance response to arsenite. This finding may help to explain why rodents do not develop tumors after exposure to arsenite, while humans do. In addition, all human cell lines tested were much more sensitive to arsenite compared to Chinese hamster cells. Human keratinocytes were especially sensitive. In general, human cells resemble arsenic hypersensitive Chinese hamster As/R27D cells, which have lost a protective mechanism found in wild-type Chinese hamster cells. PMID- 9219569 TI - Effect of hepatic methyl donor status on urinary excretion and DNA damage in B6C3F1 mice treated with sodium arsenite. AB - This study evaluated the effect of hepatic methyl donor status on the ability of sodium arsenite (2.5, 5.0 and 10.0 mg/kg) administered by gavage once or on four consecutive days to induce DNA damage in male B6C3F1 mice. Maintenance on a choline-deficient (CD) diet prior to treatment resulted in mice with hepatic methyl donor deficiency (HMDD) and altered arsenical metabolism, as demonstrated by a decreased total urinary excretion of inorganic and organic arsenicals. The alkaline (pH > 13) Single Cell Gel (SCG) assay was used to evaluate for the induction of DNA damage (single strand breaks, alkali labile sites, DNA crosslinking) in blood leukocytes, liver parenchymal cells, and cells sampled from bladder, lung, and skin, while the bone marrow erythrocyte micronucleus (MN) assay was used to assess for the induction of chromosomal damage in bone marrow cells. Treatment with sodium arsenite once or four times induced a significant decrease in DNA migration (indicative of DNA crosslinking) in bladder and liver parenchymal cells of hepatic methyl donor sufficient (HMDS) mice, but in skin cells of HMDD mice. Both HMDD and HMDS mice exhibited a significant increase in the frequency of micronucleated polychromatic erythrocytes (MN-PCE) in bone marrow following four, but not following one, treatments. However, the positive response occurred at a lower dose for HMDS mice and, in these mice, bone marrow toxicity, as demonstrated by a significant reduction in the percentage of PCE, was present also. These results indicate that hepatic methyl donors deficiency significantly decreases the total urinary excretion of orally administered sodium arsenite and markedly modulates target organ arsenic-induced DNA damage, with an apparent shift from liver and bladder to skin. PMID- 9219570 TI - Study of arsenic mutagenesis using the plasmid shuttle vector pZ189 propagated in DNA repair proficient human cells. AB - Arsenic is considered a human carcinogen and although it is non-mutagenic in bacterial or human cells, arsenic interacts synergistically with genotoxic agents in the production of mutations. To gain insight into the possible mechanisms of action of arsenic in mutagenesis we studied the effects of sodium arsenite exposure on UV mutagenesis using the pZ189 shuttle vector system in DNA repair proficient GM 637 human fibroblasts. The purpose of the study was to determine whether arsenic alone induces mutations in the supF gene and whether the combination of arsenic and UV irradiation leads to a yield of mutants greater than the sum of the arsenic or UV treatments alone. Treatment of fibroblasts for 72 h with 5.0 microM of sodium arsenite alone produced significant increases in the pZ189 mutant frequency; 1 and 2.5 microM arsenite were not mutagenic. UV irradiation (320 J/m2) increased the yield of mutants 3.5-fold above the background rate. When UV-irradiated plasmid was allowed to replicate in fibroblasts treated with 1, 2.5, or 5.0 microM arsenite, the yields of mutations were significantly greater (p < 0.01) than the yield expected if the effects of each treatment were simply additive. The greatest potentiation of UV-induced mutations (4.9-fold) was observed at 1 microM arsenite, a concentration that was neither mutagenic itself nor cytotoxic. Restriction digest and DNA sequencing analyses indicated that arsenite alone produces both large-scale rearrangements, frameshifts and base substitutions. Hotspots for deletions were observed to be associated with a previously reported deletion hotspot involving 5'-CpC and runs of cytosines. Base substitutions observed involved A:T-->T:A transversions. The results indicate that arsenite alone is mutagenic in human cells using the supF reporter gene. The pZ189 shuttle vector may provide a model to study the molecular nature of co-mutagenesis of arsenic and other environmental agents. Further characterization of arsenic's effects on DNA repair and mutational spectra may be useful in the development of molecular markers in studies of arsenic carcinogenesis in human populations. PMID- 9219571 TI - Inhibition of poly(ADP-ribose) polymerase by arsenite. AB - Inorganic arsenic is considered a human carcinogen based principally on epidemiological evidence. Unlike most initiating chemicals, arsenic is inactive or extremely weak in its ability to directly induce gene mutations. Arsenite has been shown, however, to enhance mutagenicity when present with other agents such as UV radiation. Synergistic potentiation of chromosomal damage has been shown with co-treatment with DNA-crosslinking agents. Arsenite at low concentrations is known to be highly selective in reacting with closely spaced (vicinal) dithiol groups in proteins. Poly(ADP-ribose) polymerase (PARP) is known to contain such vicinal dithiol groups. Stimulation of PARP is an immediate response of eukaryotic cells to DNA strand breaks and has been implicated in DNA repair. The effect of treatment with sodium arsenite on PARP activity was assessed as follows: Molt-3 cells (a human T-cell lymphoma-derived cell line) in culture were treated for 24 h with concentrations of sodium arsenite ranging from 2.5 up to 25 microM. Speciation of inorganic arsenic and cell viability were determined. Cell cycle kinetics were measured by flow cytometry. Poly(ADP-ribose) synthesis was assayed using a palindromic decameric deoxynucleotide to stimulate enzyme activity. Results show that arsenite decreases PARP activity in a dose-dependent manner with an approximately 50% decrease in enzyme activity at 10 microM arsenite and 80% viability. The percent of cells in S-phase increases with increasing concentration of arsenite. These results provide further indication that arsenite may potentiate genetic damage through reaction with dithiols in DNA repair proteins such as PARP, perhaps resulting in interference with normal repair function. PMID- 9219572 TI - Possible carcinogenic potential of dimethylarsinic acid as assessed in rat in vivo models: a review. AB - The modifying effects of dimethylarsinic acid (DMA), the major metabolite of ingested arsenicals in most mammals, on chemical carcinogenesis were investigated using rat in vivo models and reviewed here. In a multi-organ bioassay, rats pretreated with 5 carcinogens were administered DMA at various concentrations in their drinking water. Significantly increased tumor induction due to DMA was observed in the urinary bladder, kidney, liver, and thyroid gland. This was associated with significantly elevated ornithine decarboxylase activity in the kidneys of DMA-treated animals. To estimate the hazard levels of its promoting influence, further examinations were carried out concerned with urinary bladder and liver carcinogenesis. Doses of 25 and 50 ppm, respectively, of DMA were found capable of enhancing lesion development in the two organs. In conclusion, our data indicate that DMA is a carcinogen or promoter in the urinary bladder, liver, kidney and thyroid gland, in line with previous epidemiological findings. PMID- 9219574 TI - Types of mutations induced by glyoxal, a major oxidative DNA-damage product, in Salmonella typhimurium. AB - We have analyzed the types of mutations induced by glyoxal, a major oxidative DNA damage product, in Salmonella typhimurium. A set of six strains, TA7001 to TA7006, was used to detect base-pair substitutions, and the TA98 strain was employed to detect frameshift mutations. Glyoxal did not induce mutations at A:T base pairs. The majority of the mutations induced by glyoxal were base-pair substitutions at G:C base pairs, and a small level of frameshift mutations was detected in the TA98 strain. PMID- 9219573 TI - Mutant frequency of lacI in transgenic mice following benzo[a]pyrene treatment and partial hepatectomy. AB - The Big Blue, transgenic mouse provides an in vivo mutation system that permits the study of pharmacodynamic parameters on mutant frequency (MF) following xenobiotic exposure. We have studied the effects of cellular proliferation on the frequency of mutations in the lacl transgene by evaluating the MF in the liver of male C57B1/6 Big Blue mice following treatment with benzo[a]pyrene (B[a]P) and a partial hepatectomy. Mice received either 40 mg/kg of B[a]P in corn oil or corn oil alone by i.p. injection on three consecutive days, followed by a partial hepatectomy on the fourth day. Three days later (i.e., 7 days following the initial B[a]P injection), the animals were sacrificed and the MF in the liver was compared to the MF observed in the liver of the same mouse at the time of hepatectomy. Induction of cytochrome P-450 1A (CYP1A) following B[a]P treatment was evident by Western blot analysis. The MF in untreated control animals was not significantly different at hepatectomy (4.7 +/- 0.8 x 10(-5)) and 3 days later, at sacrifice (3.0 +/- 0.4 x 10(-5)). Neither was the MF observed in the B[a]P treated mice at the time of sacrifice (12.0 +/- 2.1 x 10(-5)) significantly different from the MF observed at the time of hepatectomy (10.6 +/- 5.3 x 10( 5)). However, B[a]P-treatment resulted in a 4.0-fold increase in MF at sacrifice which was significantly different (p < 0.05), when compared to the untreated controls. The B[a]P-treated mice at hepatectomy showed a modest 2.2-fold increase in MF which was not statistically significantly different from the untreated controls. In addition, both control and B[a]P-treated tissues gave sectored mutant plaques. The sectored plaque frequency (SPF) was significantly elevated (p < 0.05) in the B[a]P-treated mice at hepatectomy (4.2 +/- 1.0 x 10(-5)) and sacrifice (7.3 +/- 2.4 x 10(-5)) as compared to the respective frequency in the control mice at hepatectomy (1.9 +/- 0.7 x 10(-5)) and sacrifice (1.4 +/- 0.2 x 10(-5)). One explanation for this data is the persistence of the B[a]P adducts in the mouse genomic DNA that was packaged into the lambda phage, and ultimately fixed as mutations in Escherichia coli. PMID- 9219575 TI - Antimutagenic effects of dehydrozingerone and its analogs on UV-induced mutagenesis in Escherichia coli. AB - Antimutagenic activities of dehydrozingerone, benzalacetone and its analogs substituted with the hydroxyl, methoxyl or methyl group on the benzene ring were investigated by the post-treatment for UV-induced mutagenesis in Escherichia coli WP2s (uvrA). In this assay, dehydrozingerone was a poor antimutagen. Among the test compounds except for 2-hydroxybenzalacetone, benzalacetone was the most strong antimutagen, showing that the ring substitutions decrease the antimutagenic activities. 2-Hydroxybenzalacetone was, however, more effective than benzalacetone. The antimutagenicity of 2-hydroxybenzalacetone might be dependent on the property that is known to associate inter-molecularly by hydrogen bonding between the hydroxyl group and the carbonyl group. The effects of the side chain, single, double or triple bond and its adjacent carbonyl group, were further investigated using benzylacetone with saturated carbonyl chain, benzalacetone with double-bonded carbonyl chain, 4-phenyl-3-butyn-2-one with triple-bonded carbonyl chain and trans-beta-methyl styrene with only double bond. Benzalacetone and 4-phenyl-3-butyn-2-one suppressed the UV-induced mutagenesis, but benzylacetone and trans-beta-methyl styrene scarcely inhibited the mutagenesis: an alpha, beta-double or triple-bonded carbonyl system was necessary for the antimutagenic activity. 4-Phenyl-3-butyn-2-one showed the potent antimutagenicity at one order lower concentrations than that of benzalacetone. Benzalacetone, 2-hydroxybenzalacetone and 4-phenyl-3-butyn-2-one that were effective on the UV-induced mutagenesis also decreased the gamma-induced mutagenesis in Salmonella typhimurium TA2638. PMID- 9219576 TI - Isolation of an APRT heterozygote from TK6 human lymphoblasts: predominance of multi-locus loss of heterozygosity among spontaneous APRT-mutants. AB - The TK6 human B lymphoblastoid cell line contains two easily and widely used selectable markers: the X-linked, hemizygous hprt locus, and the heterozygous tk locus on chromosome 17q. In this study, rare APRT heterozygotes were directly isolated from the TK6 population by clonal selection in cell culture medium supplemented with 5 micrograms/ml of 8-azaadenine. One of nine isolated heterozygotes, AZH1, was characterized extensively. APRT- mutants can be recovered from AZH1 at a mutation rate of 1.5 x 10(-7), similar to rates previously determined for the selection of TK- and HPRT- mutants from TK6. A unique sequence alteration was identified in the non-functional aprt allele at position 1930. A G:C to A:T transition at this site alters the canonical AG splice acceptor dinucleotide in exon 3, and also results in the destruction of a Stul recognition sequence. This polymorphism was used to analyze loss of heterozygosity in a set of 32 spontaneous APRT- mutants by restriction analysis following PCR amplification. Analysis of flanking microsatellite dinucleotide polymorphisms demonstrated that LOH occurring in spontaneous APRT- mutants is nearly always a multi-locus event extending at least 7.5 cM along chromosome 16q. This pattern of LOH among APRT- mutants differs from extensive LOH in spontaneous, normal-growth TK- mutants derived from TK6 cells (p < 0.0001), and suggests that cis-acting factors may be equally important in shaping the mutational spectrum as trans-acting factors such as cellular apoptotic capacity. PMID- 9219577 TI - Dichotomous relationship between DNA reactivity and the induction of sister chromatid exchanges in vivo and in vitro. AB - Structural analyses of the determinants associated with the induction of bone marrow sister chromatid exchanges in mice indicate that the phenomenon is based on an electrophilic attack on DNA. In that respect this phenomenon is different from the basis of the induction of SCE in cultured cells or of micronuclei in the bone marrow of rodents. The latter two phenomena involve other targets as well. Based on the recognition that the vast majority of recognized human carcinogens are genotoxic, the present finding indicates that the in vivo induction of SCE is a good biomarker, possibly even a biodosimeter, for exposure to potential carcinogens. PMID- 9219578 TI - Human Rad52 protein promotes single-strand DNA annealing followed by branch migration. AB - In the yeast, Saccharomyces cerevisiae, the Rad52 gene is important for both mitotic and meiotic recombination. Homologs of the Rad52 gene have been identified in several eukaryotic organisms, ranging from yeast to man. As reported here, human Rad52 protein binds to both single- and double-stranded DNA; and acting on a pair of single-stranded and partially duplex substrates it promotes annealing of complementary strands of DNA, which is followed by branch migration. PMID- 9219579 TI - Mutation load and human longevity. AB - Since paternal age at reproduction is considered to be the main factor determining human spontaneous mutation rate (Crow, J. (1993) Environ. Mol. Mutagenesis, 21, 122-129), the effect of paternal age on human longevity was studied on 8,518 adult persons (at age 30 and above) from European aristocratic families with well-known genealogy. The daughters born to old fathers (50-59 years) lose about 4.4 years of their life compared to daughters of young fathers (20-29 years) and these losses are highly statistically significant, while sons are not significantly affected. Since only daughters inherit the paternal X chromosome, this sex-specific decrease in daughters' longevity might indicate that human longevity genes (crucial, house-keeping genes) sensitive to mutational load might be located in this chromosome. PMID- 9219580 TI - Chromosome aberrations in Syrian hamsters following very low radiation doses in vivo. AB - This paper addresses a report of a large increase (approximately 6- to 11-fold) in chromosome aberrations in lymphocytes of persons in Salzburg attributed to their exposure to fallout from the Chernobyl cloud. Their additional exposure, approximately 0.3 mGy in 1 year, comprised about a 30% increase in their normal background radiation dose. The report has attracted considerable attention because, if correct, it seriously challenges assumptions of linearity in the low dose response for chromosomal damage and, by implication, the linear, no threshold hypothesis for risk of induced cancer. An experiment has been carried out with Syrian hamsters treated with caesium-137 to produce a range of doses comparable with those calculated for the persons in Salzburg. No significant elevation in lymphocyte aberration yields was found in the hamsters, thus arguing against the conclusions of the Salzburg study. PMID- 9219581 TI - Sister chromatid exchange induction in sheep peripheral blood mononuclear cells by helio-neon laser radiation. AB - The effects of laser light on the cellular DNA have not been extensively characterized. Low-power laser sources, such as the helium-neon (He-Ne) laser with a wavelength of 632.8 nm, have been found to produce photobiological and photodamage effects with evidence of interference with cell replication. We have investigated the effects of He-Ne laser irradiation on sister chromatid exchange (SCE) frequencies in sheep peripheral blood mononuclear cells (PBMC). Cultured cells were irradiated once at 6 selected energy intensities of laser irradiation and then stimulated with pokeweed mitogen and cultured in the presence of 5 bromodeoxyuridine (BrdUrd). The frequency of SCEs of both irradiated and non irradiated cells were analyzed. The mean SCE of irradiated cells significantly increased with growing energy density up to a laser dose of 24 J/cm2, whereas after an energy density of 24 J/cm2, the SCE frequency decreased with increasing energy densities. We concluded that the application of He-Ne laser irradiation at energy densities ranging from 2 to 96 J/cm2 produced a different effect on SCE frequency in sheep PBMC in vitro. PMID- 9219582 TI - Induction of micronuclei, hyperdiploidy and chromosomal breakage affecting the centric/pericentric regions of chromosomes 1 and 9 in human amniotic fluid cells after treatment with asbestos and ceramic fibers. AB - This article describes the induction of micronuclei, hyperdiploidy and chromosome breakage in human amniotic cells in vitro by amosite, chrysotile and crocidolite asbestos, and ceramic fibers. The response of human (amniotic fluid cells) and rodent (Syrian hamster embryo fibroblasts, SHE) cells to fiber treatment was compared using the micronucleus assay. The data of the rodent studies were taken from a previous investigation (Dopp, E. et al. (1995) Environ. Health Perspect., 103, 268-271). All types of mineral fibers caused a significant increase of micronucleated cells. The kinetochore analysis revealed that all three types of asbestos and ceramic fibers yielded similar effects. Approximately 50% of the induced micronuclei were kinetochore-negative indicating formation through clastogenic events. Human amniotic cells were much less susceptible than SHE cells to the induction of micronuclei by mineral fibers. This again demonstrates that SHE cells are more susceptible to chromosomal changes than human amniotic fluid cells. The application of fluorescence in situ hybridization (FISH) with tandem DNA probes yielded more detailed information about specific structural chromosome aberrations in the 1 (cen-q12) and 9 (cen-q12) regions and about abnormal numbers of chromosomes in interphase human amniotic fluid cells. Using this FISH approach we found a statistically significant increase of chromosomal breakage in the pericentric heterochromatin regions of chromosomes 1 and 9 in interphase human amniotic cells after exposure to asbestos and ceramic fibers compared to control cells. The number of hyperdiploid cells was also significantly increased. Our results show that asbestos fibers as well as ceramic fibers are inducers of structural and numerical chromosomal aberrations in human amniotic fluid cells. PMID- 9219583 TI - Increase of chromosomal aberrations induced by ionising radiation in peripheral blood lymphocytes of civil aviation pilots and crew members. AB - This study inquires if there is an increase of chromosomal aberrations by ionising radiation of cosmic origin in civil pilots and flight-crew members. Totals of 37,208 exposed cells and 10,950 control cells, from 192 and 55 donors respectively, were scored averaging 200 observations per subject. The analysis showed the increase of dicentric and ring chromosomes in peripheral blood lymphocytes of the flight personnel. The difference of the total aberration frequencies between exposed and control was statistically significant. PMID- 9219584 TI - Detection of DNA damage in human lymphocytes by alkaline single cell gel electrophoresis after exposure to benzene or benzene metabolites. AB - The alkaline single cell gel electrophoresis (Comet) assay was applied to study the occurrence of DNA damage in peripheral lymphocytes of human subjects with occupational exposure to low levels of benzene (twelve gasoline station attendants, with average benzene exposure of 0.3 mg/m3, 8 h TWA). The results obtained show a significant excess of DNA damage in lymphocytes of exposed workers, compared to matched unexposed controls (p = 0.028, Mann-Whitney U-test). Averaged tail moment values, based on 100 cells/individual, were 1.900 microns in the exposed and 0.936 micron in the unexposed group. In addition, exposed subjects showed a clearcut excess of heavily damaged cells, with tail moments > 90th percentile of the overall distribution (13.5 vs. 6.5%, p = 0.013, Mann Whitney U-test). No correlation was found between the extent of DNA damage and the ages or smoking habits of the subjects. In order to assess the plausibility of the involvement of benzene in the results of the ex vivo study, further experiments were performed treating in vitro peripheral lymphocytes from unexposed donors with benzene metabolites hydroquinone, benzoquinone and benzenetriol. In these experiments, all benzene metabolites exerted a marked effect on resting lymphocytes, the lowest effective concentrations being below 1 microgram/ml. Conversely, far greater concentrations were required for the induction of significant DNA damage in parallel experiments with hydroquinone on mitogen stimulated lymphocytes. Addition of the DNA repair inhibitor cytosine arabinoside (Ara-C, 1-10 micrograms/ml) partially restored the sensitivity of stimulated cells to hydroquinone, an indication of the active processing of induced DNA lesions in growing cells. These results are discussed also in relation to the role of peripheral lymphocytes as target tissue in the biomonitoring of human exposure to genotoxic agents. PMID- 9219585 TI - Correlation between micronuclei induction and cell survival in V79 cells exposed to paclitaxel (taxol) in conjunction with radiation. AB - Exposure of V79 cells to different doses of gamma-irradiation resulted in a dose dependent decline in their survival. The treatment of cells with paclitaxel before irradiation resulted in a further decline in the cell survival. Conversely, the micronuclei frequency increased with the increasing dose of radiation in both irradiated and paclitaxel + irradiated cultures at all the three post-irradiation time periods studied. However, no significant difference among the frequencies of micronuclei was found at 16, 22 and 28 h post-exposure. The trend for cell proliferation was similar to that of cell survival. The cell proliferation declined with increasing dose of radiation. The paclitaxel treatment further reduced cell proliferation compared to the irradiated control group. The dose response for all the parameters for both groups was linear quadratic. The biological response between micronuclei induction and cell survival was also determined. The cell survival and micronuclei formation were inversely related and the correlation between cell survival and micronuclei formation (biological response) was linear quadratic for both irradiated and paclitaxel + irradiated groups. PMID- 9219586 TI - The effects of maternal cigarette smoke exposure on somatic mutant frequencies at the hprt locus in healthy newborns. AB - We utilized the hprt T-cell cloning assay to prospectively determined the somatic mutant frequency at the hprt locus of fetal T-lymphocytes exposed in utero to maternal active and passive cigarette smoke. In addition, a maternal questionnaire was administered to evaluate a number of social and medical parameters that may effect hprt mutant frequency. Newborn cord blood plasma cotinine levels were determined on all subjects to compare in utero tobacco metabolite levels with maternal smoking histories. A total of 63 newborns were enrolled and placed into four groups: Group I (n = 21), newborns whose mothers had no history of active or passive cigarette exposure during the pregnancy; Group II (n = 12), newborns whose mothers actively smoked cigarettes throughout the pregnancy; Group III (n = 8), newborns whose mothers actively smoked cigarettes during first trimester only; and Group IV (n = 22), newborns whose mothers were exposed only to passive cigarette smoke. Our analysis showed no statistically significant difference in hprt mutation frequency between any of the four groups. A significant increase in plasma cord blood cotinine was detected in Group II, newborns whose mothers were active cigarette smokers throughout the pregnancy. Our data indicate that exposure to active and passive maternal cigarette smoke in utero does not result in a significant increase in somatic mutant frequency as determined by the hprt T-cell cloning assay. PMID- 9219587 TI - 'Spontaneous' genetic damage in man: evaluation of interindividual variability, relationship among markers of damage, and influence of nutritional status. AB - The 'spontaneous' frequency of genetic damage (normal background) and the possible relationship of this damage to nutritional variables in humans were investigated in 22 subjects using several indices of genetic damage. The subjects were chosen, out of 122 initially analyzed, for being at the extremes of the highest and lowest values of one index of genetic damage, the frequency of micronucleated erythrocytes in peripheral blood. This index reflects chromosomal damage and loss in bone marrow erythropoietic cells. The assay for micronuclei is convenient but is restricted to splenectomized individuals because the human spleen removes micronucleated cells. The initial 122 subjects were splenectomized, but all were normal and healthy at the time of this study and none had a previous history of neoplastic disease. Factors investigated were stability of micronucleus frequency as a function of time, correlations among multiple markers of genetic damage, and influence on damage indices of nutritional variables, including blood levels of folate, B12 and antioxidant vitamins. Among different individuals, the range of values was 10-fold or more in the erythrocyte micronucleus, glycophorin A, plasma ascorbate and urinary 8 hydroxydeoxyguanosine (oxo8dG) assays, was approximately 6-fold in the lymphocyte micronucleus assay, and was 2-fold in the lymphocyte sister chromatid exchange (SCE) assay. Red blood cell folate and plasma folate, B12 and alpha-tocopherol values varied by up to 10-fold among individuals. Micronucleus frequencies in erythrocytes and peripheral blood lymphocytes ranged from < 0.3 to 16.9/1000 in mature red blood cells, < 1 to 33/1000 in reticulocytes, and 2.5 to 15/1000 in binucleate lymphocytes. Frequencies of glycophorin A variant erythrocytes ranged from 5.6 to 77.3 x 10(6) N/0 cells and 3.2 to 16.2 x 10(6) N/N cells, and oxo8dG excretion varied from 32 to 397 pmol/kg/day. Although a wide range of values was observed in each genetic endpoint, the extreme values for various endpoints of genetic damage were not observed in the same individuals. The frequency of micronucleated erythrocytes varied over time within individuals and indicated that individuals with the highest levels of damage exhibit greater variability than those with lower levels. In some subjects, frequencies of micronucleated erythrocytes changed dramatically over an interval of 2-3 years: four subjects with initial micronucleated reticulocyte frequencies of 20.4, 5.9, 6.4 and 33/1000 changed to 2.5, 20.5, 18.5 and 12/1000, respectively. Among more than 150 individuals we have studied, including the 64 individuals studied by Everson et al. [(1988) J. Natl. Cancer Inst., 80, 525-529] and Smith et al. [(1990) Cancer Res., 50, 5049-5054], the seven individuals with the highest observed frequencies of micronucleated erythrocytes all had exceptionally low values of plasma folate, red cell folate, or plasma B12, suggesting that folate and B12 status are the major determinants of the types of damage that lead to spontaneous micronucleus formation in erythrocytic cells. PMID- 9219588 TI - In vivo antioxidant status, DNA damage, mutation and DNA repair capacity in cultured lymphocytes from healthy 75- to 80-year-old humans. AB - The accumulation of damage to cellular biomolecules, including DNA, over time may play a significant role in the aetiology of the ageing process. We have previously quantified DNA damage and mutation within cultured lymphocytes from healthy human male subjects in three different age groups (35-39, 50-54 and 65-69 years). The results of that study showed an age-related increase in DNA damage and mutations in lymphocytes. In addition, an age-related decrease in the capacity of the lymphocytes to repair H2O2-induced DNA damage was found. In this article, we report the findings of an extension to the earlier study. Thirty-one generally healthy male and female subjects between the ages of 75 and 80 years were recruited. Using a number of bioassays, we were able to determine; basal levels of DNA damage (for 18 subjects) and mutant frequency at the hypoxanthine phosphoribosyltransferase (hprt) gene locus (for 16 subjects) within cultured lymphocytes. In addition, in vivo antioxidant status (for all study subjects) and the capacity of lymphocytes to repair H2O2-induced DNA damage (for 18 subjects) were also assessed. The results obtained showed: that the mean basal level of DNA damage in lymphocytes from subjects in the 75- to 80-year age group (12.6 +/- 4.7%) was similar to that of the 35- to 39-year age group (13.3 +/- 3.3%), p = 0.42 (Mann-Whitney); there was no significant difference between log mean mutant frequency at the hprt gene locus in lymphocytes from the 75- to 80-year age group (0.31 +/- 0.33) compared to that observed in the 35- to 39-year age group (0.24 +/- 0.21; Student's t-test, t = 0.68, p > 0.05). Levels of the antioxidants glutathione peroxidase (GPx EC 1.11.1.9), catalase (CAT; EC 1.11.1.6) and caeruloplasmin (CPL; EC 1.16.3.1) were significantly elevated in the 75- to 80 year age group, compared to the 35- to 39-, 50- to 54- and 65- to 69-year age groups. Levels of bilirubin (BR) were reduced in the 75- to 80-year age group, the decrease being contributed by the female subjects. No differences in levels of superoxide dismutase (SOD; EC 1.15.1.1) or uric acid (UA) were found between the 4 age groups. Following treatment of lymphocytes with H2O2, we did not find any difference in the susceptibility of lymphocytes to DNA damage in the 75- to 80-year age group, compared to the other age groups. The DNA repair capacity in lymphocytes from individuals in the 75- to 80-year age group was similar to that of the 35- to 39-year age group, for all time points assessed. These results highlight the importance of DNA repair processes and antioxidant defence systems for maintaining genomic stability in vivo. PMID- 9219589 TI - Expression of the Pseudomonas aeruginosa uvrA gene is constitutive. AB - By complementation of an Escherichia coli uvrA-defective mutant, the uvrA gene of Pseudomonas aeruginosa has been isolated from a genomic library. The nucleotide sequence of the promoter and the 5'-coding region of this gene have been determined. A P. aeruginosa SOS consensus region, which functions as a binding site for the LexA repressor molecule, was not present in the 629-bp upstream region of the cloned uvrA gene. Analysis of transcriptional fusions with the lacZ gene demonstrates that, contrary to what happens with its homologous gene of E. coli, the P. aeruginosa uvrA gene is not DNA damage-inducible. However, the UvrA protein must be functional in P. aeruginosa cells because an uvrA-defective mutant is extremely sensitive to UV radiation. PMID- 9219590 TI - [Computerized tomography-guided location of atypical small lung nodules]. PMID- 9219591 TI - The functional damages of ischemic/reperfused skeletal muscle. AB - Skeletal muscle is frequently damaged by ischemia-reperfusion when exposed to direct injury or in the surgical practice. The purpose of the present experiments was to examine how the different types of skeletal muscles (fast & slow) react functionally to one and two hours of ischemia followed by two weeks of reperfusion. The fast-twitch (m. extensor digitorum longus/EDL) and the slow twitch (m. soleus/SOL) muscle were prepared. They were stimulated, in vivo, either directly or indirectly at different reperfusion times following tourniquet ischemia, and the contraction force (CF) was recorded. CF of the EDL was reduced over 40% and 90% of the control value during the first 24 hours of reperfusion after 1 and 2 hours of ischemia, respectively. It was about 50% at the end of the 2nd week in the one-hour group. CF increased significantly during the second week if ischemia lasted for two hours. Reduction of CF in the SOL muscle was over 50% and 90% following one and two hours of ischemia, respectively. It further decreased in the 1-hour group, and it started to regenerate from the second week after 2 hours of ischemia. It is concluded that 1. two hours of ischemia causes significantly more severe damages in both types of skeletal muscles than one hour. 2. There is a reperfusion injury in both muscles during the first week of reperfusion. 3. The two types of muscles regenerate differently, i.e. the SOL starts to regenerate earlier than the EDL. PMID- 9219592 TI - Effects of midazolam on blood glucose fibrinolysis-serum lipids in normoglycemic normolipidemic rats. PMID- 9219593 TI - Ipriflavone metabolite-III inhibits LPS-induced nitric oxide release from RAW 264.7 cells. AB - Ipriflavone [CAS No.: 35212-22-7] is a novel drug used in the treatment of osteoporosis successfully. However, its mechanism of action has not been fully clarified yet. We investigated the effects of ipriflavone and its metabolites (I, II, III, V, VI, VII) on lipopolysaccharide (LPS)-induced nitric oxide (NO) release from RAW-264.7 mouse macrophage cells. Our data show that the LPS-induced NO release from RAW-264.7 cells was significantly inhibited by ipriflavone metabolite-III [7-isopropoxy-3-(4-hydroxyphenyl)-4H-1-benzopyran-4-one], [CAS No.: 97846-18-9] in a dose (3 x 10(-8)-10(-5) M)-dependent manner. Ipriflavone itself and its other metabolites had much lower inhibitory effect on the LPS induced NO release from RAW-264.7 cells. The IC50-value of ipriflavone metabolite III (1.0 x 10(-6) M) was between the IC50-values of the two reference compounds dexamethasone (4.0 x 10(-8) M) and NG-Nitro-L-arginine (L-NNA, 7.5 x 10(-5) M). Our finding suggests that some of the beneficial effects of ipriflavone in the treatment of osteoporosis might be mediated by its metabolite-III. PMID- 9219594 TI - Regional heterogeneity and differential vulnerability of cerebral and spinal vascular CO2-responsiveness during graded haemorrhagic hypotension. AB - Regional inhomogeneity of cerebrovascular CO2-sensitivity as well as its changes at three different levels of standardized haemorrhagic hypotension were studied in ten distinct brain and spinal cord regions of anesthetized, ventilated cats. Regional cerebral blood flow was measured with radiolabelled microspheres in hypocapnic, normocapnic, and hypercapnic conditions, and CO2-responsiveness was determined from the equation of the slopes of the best fit regression lines to the obtained flow values. It was concluded that in normotensive, normoxic cats response of the cerebral and spinal vessels to PaCO2 alterations can be assigned to four major categories. The CO2-responsiveness of a brain region is not solely determined by the rate of its basal steady state blood flow: CO2-reactivity of the hypothalamus was significantly different from that of any other investigated regions with almost identical steady state flow values. Vulnerability of the cerebrovascular CO2-sensitivity during hypotension was different from region to region, with the vessels of the pons-medulla oblongata region being the most sensitive to haemorrhage. Reduced regional cerebral and spinal CO2-responsiveness during haemorrhage is not a consequence of a reduced L-arginine supply for nitric oxide generation since administration of an excess amount of the precursor L arginine failed to restore the haemorrhage-induced reduction of regional CO2 sensitivity at the 60 mm Hg mean arterial pressure level. PMID- 9219595 TI - Prevention of indomethacin-induced acute gastric mucosal injury by spermine in the rat. AB - The effect of spermine tetrahydrochloride was investigated on the indomethacin induced acute mucosal injury and mucosal lipid peroxidation in rats. Spermine was given orally at doses of 25, 75 and 150 mg/kg and, intraperitoneally at 10, 25 and 50 mg/kg 1 hour before 25 mg/kg indomethacin administration. Macroscopic injury was measured by the use of a stereomicroscope. Mucosal malondialdehyde (MDA) levels were determined by an HPLC method. Oral spermine reduced dose dependently the extent of macroscopic injury from 22.87 +/- 2.88 mm to 11.8 +/- 2.48 mm (p < 0.01, 25 mg/kg), 6.53 +/- 4.19 mm (p < 0.0007, 75 mg/kg) and 0.25 +/ 0.25 mm (p < 0.0001,150 mg/kg). Intraperitoneally administered spermine prevented dose-dependently the indomethacin produced lesions from 19.25 +/- 4.31 mm to 12.63 +/- 3.18 mm (p < 0.01, 25 mg/kg) and to 0.33 +/- 0.33 mm (p < 0.001, 50 mg/kg). Doses necessary for intraperitoneal protection were lower than those achieving similar effect, orally suggesting different mechanisms of action. Neither oral or intraperitoneal spermine influenced the mucosal MDA levels. PMID- 9219596 TI - ETB receptor mediating pulmonary hypertension and bronchoconstriction induced by endothelin-1 in the guinea pig. AB - Endothelin (ET-1) caused dose-related contraction of isolated superfused bronchus and pulmonary artery and bronchoconstriction and pulmonary vascular hypertension of the heart lung preparation (HLP) of guinea pig. The specific ETA receptor antagonist BQ 123 completely blocked the responses of the pulmonary artery, but failed to affect those of bronchus and of HLPs. The specific ETB receptor agonist Sarafotoxin S6c caused contractions of bronchus, but not of pulmonary artery, and bronchoconstriction and pulmonary hypertension in HLPs. It is concluded that non ETA subtype receptors, perhaps ETB, appear to be the main responsible for the potent pulmonary hypertensive effects of ET-1. PMID- 9219597 TI - Effect of granulocyte colony-stimulating factor on granulopoiesis of congenital neutropenic children. AB - We report on two patients with congenital neutropenia, who were treated with filgrastim (recombinant human granulocyte-colony stimulating factor, G-CSF). A poor growth of bone marrow colonies and low sensitivity of colony forming units to colony stimulating factor in vitro before treatment seemed to be associated with a requirement for higher doses of G-CSF to achieve good clinical response in vivo. PMID- 9219598 TI - Changes in the biliary excretion of exogenous organic anions by streptozotocin induced diabetes. AB - A transient depression of blood-glucose level was found after streptozotocin administration, which can be explained by insulin release due to the destroying effect of streptozotocin on beta-cells of pancreas, the biliary flow was elevated, when blood sugar level was low. After the transient decrease the blood glucose level was elevated and remained in a diabetic range (300-500 mg/100 ml). During the diabetic period a biphasic change was observed in the biliary flow: 23 48 hours after streptozotocin injection a depression, however, 5-10 days after streptozotocin administration an elevation was detected in the bile flow. Changes found in biliary excretion rate of exogenous organic anions were parallel with those of biliary flow. PMID- 9219599 TI - Effect of glimepiride and glibenclamide, inhibitors of ATP-dependent K(+) channel, on ischaemia-reperfusion induced arrhythmias in rats. AB - Glibenclamide or glimepiride pretreatment (5 mg/kg i.p. 30 min prior to coronary ligation) significantly improved the survival rate during reperfusion after 6 min myocardial ischaemia in rats (82% or 67% vs. 17% in controls). Smaller dose of glibenclamide (0.01 mg/kg) did not influence the survival rate (31%), while glimepiride still offered a significant protection (61%). These results suggest that different KATP inhibitors may increase the chance to survive life threatening arrhythmias during myocardial ischaemia-reperfusion. PMID- 9219600 TI - rho-Aminophenylalanine6 angiotensin II (AII) improves learning and memory in rats. AB - All and rho-aminophenylalanine6 angiotensin II, given i.c.v. (1 nmol) enhanced memory and stereotypic behaviour leaving gross motor activity unchanged. Both peptides showed also some anxiogenic action. PMID- 9219601 TI - The NMDA and GABA-A receptors in behavioral activity of rats. AB - The effects of bicuculline (0.25 mg/kg ip) and AP-7 (5 nmols icv) on the processes of retrieval, consolidation of conditioned reflexes, object recognition and locomotor activity were tested in rats. AP-7, bicuculline and AP-7 with bicuculline increased (but not significantly) locomotor and exploratory activity in the open field test. Only coadministration of AP-7 with bicuculline facilitated retrieval of passive avoidance in rats, but was without effect on consolidation in this test. AP-7 or bicuculline also did not influence on consolidation, when they were given alone. Object recognition was impaired (but not significantly) in groups of rats treated with bicuculline and bicuculline with AP-7. PMID- 9219602 TI - Losartan does not influence the blood platelet aggregation in normotensive rats. AB - Platelet aggregation was studied in PRP upon stimulation with ADP and collagen in normotensive rats treated with losartan (10 mg/kg). The acute and subchronic (5 days) losartan administration did not change the aggregating response of rat platelets. Similarly, in vitro study aggregation of platelets remained unaltered following incubation with losartan and its active metabolite EXP3174. In this study we presented the lack of influence of losartan and its main metabolite on rat platelet aggregation in normotensive rats. PMID- 9219603 TI - The non-peptide tachykinin NK1- and NK2-receptor antagonists SR 140333 and SR 48968 prevent castor-oil induced diarrhea in rats. AB - Castrol-oil induced diarrhea in rats was potently prevented by compounds SR 140333 and SR 48968, the first a tachykinin NK1- and the second a NK2-receptor antagonist. SR 48968 was more effective and also reduced fecal water content. PMID- 9219604 TI - Long-lasting treatment with adenosine receptor antagonists: effects on hypoxic tolerance and vascular responsiveness. AB - In electrically driven myocardial preparations obtained from chronically methylxanthine-[aminophylline (APH) and 8-phenyltheophylline (8-PT)] or solvent(DMSO)-treated guinea pigs no differences were found in alteration of mechanical activity under hypoxia and reoxygenation. The vasoconstrictor effects observed after in vitro exposure of pulmonary arterial preparations (excised from either methylxanthine- or solvent-treated guinea pigs) to both noradrenaline and PGF2 alpha were also similar. In methylxanthine-treated vascular tissues, however, nitroglycerin and NO exerted more pronounced vasorelaxant effect than in specimens prepared from solvent-treated guinea pigs. PMID- 9219605 TI - Longevity treatment with (-)deprenyl in female rats: effect on copulatory activity and lifespan. AB - Six months old ovariectomized female rats (n = 9) were treated with (-)deprenyl in a dose of 0.25 mg/kg s.c. three times a week, and (n = 9) with physiologic saline (0.1 ml/100 g) till decay. It was found that control females (n = 9) decayed within the age of fifteen months while the members of the (-)deprenyl treated group were all alive at that age. Moreover three (-)deprenyl treated female rats reached 36 months of age. Sexual activity was quite absent in both groups. The data suggests that (-)deprenyl extended the lifespan of female rats only in total absence of gonadal hormones and sexual activity. PMID- 9219607 TI - Effect of almokalant a specific inhibitor of IKr on myocardial ischaemia reperfusion induced arrhythmias in rabbits. AB - The antiarrhythmic effect of almokalant, a new type III antiarrhythmic agent, was examined by occluding and releasing the left circumflex coronary artery for 10 min, respectively, in openchest, pentobarbital-anaesthetized albino rabbits. Almokalant pretreatment increased the number of animals developing no arrythmias (5/9 vs. 1/12 in controls), and decreased the incidence of ventricular fibrillation (1/9 vs. 9/12) during reperfusion. According to our results almokalant can protect the heart against arrhythmias induced by ischaemia and reperfusion. PMID- 9219606 TI - Z1046: biochemical characterization of its dopaminergic activity. AB - Affinity of Z1046 for dopamine receptor subtypes, its ability to modulate D1- and D5-mediated AC stimulation and D1-induced cAMP accumulation were evaluated. On D1 like receptors Z1046 and fenoldopam (fen) showed a similar high affinity, being more potent than DP-5,6-ADTN and 5,6-ADTN. For the D2-like receptors, the affinity rank orders were: D2: Z1046 > or = DP-5,6-ADTN > fen = 5,6-ADTN; D3: Z1046 > DP-5,6-ADTN > fen = 5,6-ADTN; D4: Z1046 = DP-5,6-ADTN > fen = 5,6-ADTN. In AC studies the rank order was: Z1046 = fen > DP-5,6-ADTN > 5,6-ADTN. Z1046 was more efficient than fen in stimulating cAMP accumulation. These results make Z1046 an innovative agent combining D1-like and D2-like activities. PMID- 9219608 TI - Z1046: a new specific peripheral dopaminergic compound. AB - It is well established that peripheral dopamine receptors activation evokes vasodilation and neurohormonal modulation. Z1046 is a potent mixed dopaminergic agonist and is highly selective over adrenergic and serotoninergic (5-HT2) activities. In contrast, dopamine had agonist activities on all adrenergic receptors while it is known that dopexamine has a beta 2 agonist activity and is an inhibitor of the neuronal uptake. As far as fenoldopam is concerned, selective stimulation of D1-like receptors leads to an increase of renin release. In conclusion, Z1046 is a potent and specific drug for dopamine receptors. This profile makes Z1046 different from other dopaminergic agents. PMID- 9219609 TI - Peripheral serotonergic mechanisms in the cardiovascular system of epidermoid lung cancer patients. AB - The aim of the work was to evaluate the peripheral serotonergic mechanisms in patients with epidermoid lung cancer. The study was performed on lung cancer patients diagnosed on the basis of histopathological findings. The subjects were treated by radiotherapy. Whole blood 5HT in patients with epidermoid lung cancer was increased, whereas uptake of 3H-5HT by platelets was diminished in these patients when compared to control group. Radiotherapy caused a lowering of serotonin in blood to levels observed in healthy subjects. Platelet aggregation induced by ADP was diminished in patients with epidermoid lung cancer, however radiotherapy resulted in an increased sensitivity of platelets to ADP (an enhanced aggregation). On the other hand, serotonergic amplification of ADP induced platelet aggregation was higher in these patients. This effect was inhibited by radiotherapy. On the basis of our results, we may suggest that peripheral serotonergic mechanisms play an important role in pathogenesis and course of epidermoid lung cancer in humans. PMID- 9219610 TI - Effect of hyperglycemia on the intestinal elimination of p-nitrophenol in the rat. AB - Effect of hyperglycemia on the intestinal elimination of p-nitrophenol has been investigated in rats. Hyperglycemia was produced by a continuous i.v. infusion of glucose, p-nitrophenol was used as a model compound for the investigation of intestinal metabolism and excretion of drugs. Intestinal conjugation of p nitrophenol with glucuronic acid did not change significantly in hyperglycemic rats, however, formation of sulfoconjugate of p-nitrophenol was enhanced by hyperglycemia. Sum of metabolites (p-nitrophenol glucuronide and sulfate) appeared in the intestinal lumen in hyperglycemic rats was similar to the total luminal appearance of these metabolites of control rats. PMID- 9219611 TI - Do sodium nitroprusside and L-NAME affect pyrogen fever in rabbits? AB - Thermoregulatory responses after treatment with nitric oxide (NO) donor, sodium nitroprusside (SNP-3 mg/kg/h), or NO synthase inhibitor, NG-nitro-L-arginine methylester (L-NAME-100 mg/kg) were investigated in febrile rabbits (lipopolysaccharide E. coli-1 meg/kg). Pretreatment with SNP attenuated pyrogen fever as well as metabolic rate. L-NAME also inhibited postpyrogen increases in metabolism; however, this effect did not lead to antipyresis. PMID- 9219612 TI - Pharmacological analysis of nerves supplying the oesophagus. AB - Morphological studies indicate that cholinergic, peptidergic and "nitrergic" (nitric oxide releasing) nerves supply the oesophageal muscle. In the present in vitro study, intramural nerves of guinea pig and human oesophagus were activated by means of the ganglion stimulating drug nicotine and electrical field stimulation (EFS) in organ bath experiments. In general, stimulation-induced primary contractions were diminished by atropine, while non-adrenergic, non cholinergic (NANC) relaxations and after-contractions were inhibited by the NO synthase inhibitor NG-nitro-L-arginine (L-NNA). Thus we obtained pharmacological evidence for cholinergic muscarinic, as well as nitric oxide mediated mechanisms in the functional innervation of the oesophageal muscle. PMID- 9219613 TI - Nitric oxide activates an iberiotoxin-sensitive potassium channel in human saphenous vein. AB - Synthetic iberiotoxin (IBTX), an inhibitor of large conductance calcium-activated potassium channel (BKCa), was used to study the possible involvement of a specific hyperpolarizing ion channel in nitric oxide (NO)-induced venodilation. Serotonin (0.125 microM)-induced contraction of isolated human saphenous vein was dose-dependently relaxed with 50-1550 nanoM exogenous NO. 30 min preincubation of venous preparations with 90 nanoM IBTX decreased vasorelaxation induced by NO. In conclusion, a hyperpolarizing potassium channel, BKCa, may be involved in the venodilator effect of endogenous NO. PMID- 9219615 TI - Long-term ischaemic preconditioning of the heart induced by repeated beta adrenergic stress. AB - In the present study we tested the preconditioning effect of repeated beta adrenergic stress induced by Isoproterenol in chronically instrumented, conscious rabbits. We have found that at least 5 intravenous administrations of Isoproterenol, repeated at 10 min intervals, were necessary to induce a long-term cardiac adaptation manifested by a significant reduction of the harmful ischaemic changes due to cardiac stress 24 and 48 hours after preconditioning. These results suggest that a well-defined threshold level of the preconditioning stress is needed to trigger induction of metabolic changes leading to development of delayed and long-term cardiac adaptation. PMID- 9219614 TI - Electrophysiological characteristics of ventricular muscles from diabetic hearts. PMID- 9219617 TI - The enhancement and the inhibition of noradrenaline-induced cyclic AMP accumulation in rat brain by stimulation of metabotropic glutamate receptors. AB - The actions of several metabotropic glutamate receptor agonists and antagonists on noradrenaline (NA)-stimulated [3H]-cyclic AMP accumulation were investigated in rat cerebral cortical slices. Quisqualate (QUIS), L-2-amino-3 phosphonopropionic acid (L-AP3) and glutamate (GLU) elicited concentration dependent inhibition of (NA)-stimulated [3H]-cyclic AMP accumulation. In contrast (2S,3S,4S)-alpha-(Carboxy-cyclopropyl)glycine (L-CCGI), 1-Aminocyclo-pentane 1S,3R-dicarboxylate (1S,3R-ACPD), ibotenate (IBO) and (RS)-4-carboxy-3-hydroxy phenylglycine (CHPG) elicited a concentration-dependent enhancement of NA stimulated [3H]-cyclic AMP accumulation. A putative mGluR antagonist-L-AP3, inhibited the 1S,3R-ACPD-induced enhancement of the action of NA on [3H]-cyclic AMP accumulation in a biphasic manner with an IC50 of 4.5 microM for the high affinity site, which represented 65% of the total and an IC50 of 283 microM for the low affinity site. PMID- 9219616 TI - Effect of restacorin on the early and delayed after depolarization in dog Purkinje fibres. AB - The effect of restacorin and flecainide on the early afterdepolarization (EAD) and on the delayed afterdepolarization (DAD) was studied in dog cardiac Purkinje fibers by applying the conventional microelectrode technique. Restacorin and flecainide at 5 microM concentration partially abolished EAD. Further increase of the concentrations of the drugs to 10 microM EADs were completely abolished. The amplitude of DADs were significantly decreased by both restacorin (9.9 +/- 2.1 mV v.s. 2.9 +/- 1.8 mV, p < 0.01, n = 5) and flecainide (11.1 +/- 1.3 mV v.s. 0.6 +/ 0.6 mV, p < 0.01, n = 7). These results suggest that the well-established antiarrhythmic effect of restacorin and flecainide at least partially may be explained by their beneficial action against triggered abnormal automaticity. PMID- 9219618 TI - Intracellular Ca2+ imaging of cultured chicken telencephalic cells: characterization of ionotropic glutamate receptor activation. AB - In the present study we investigated the intracellular [Ca2+] increasing effect of the excitatory amino acid agonists kainate, AMPA and NMDA on fura-2/AM loaded chicken telencephalic cells in various conditions. Kainate (110 microM) increased [Ca2+]i to 256 +/- 23% of the basal level (n = 7). In Ca(2+)-free medium the effect of kainate on intracellular Ca2+ was completely abolished indicating that the primary source of the Ca2+ signal was the extracellular pool. Voltage dependent Ca2+ channel antagonism by Cd2+ decreased the intracellular Ca2+ elevation caused by 100 microM kainate indicating the involvement of voltage dependent Ca2+ channels (VDCC). PMID- 9219619 TI - Effect of ethyldeoxyuridine on 5-fluorouracil-induced neutropenia. AB - It is supposed that the toxic effect of 5-fluorouracil (5FU) on tumour cells may be increased by pretreatment with ethyldeoxyuridine (EDU). We studied the effect of this combination on neutrophil count in mice. Our present studies demonstrated that the neutropenia induced by 5 x 20 mg/kg 5FU became more severe when each dose of 5FU was preceded by 200 mg/kg EDU. PMID- 9219620 TI - Kainate-induced inhibition of voltage-activated potassium currents in cultured chick telencephalic neurons. AB - Kainate indirectly produces an inhibition of voltage-activated outward potassium currents on cultured chick telencephalic neurons, besides directly evoking activation of AMPA/kainate receptors (current with linear I-V curve, reversal potential near 0 mV). The former effect is also mediated by AMPA/kainate receptors. The role of [Ca2+]i in the inhibition was investigated by whole cell- and nystatin-perforated patch-clamp technique. Elevation of [Ca2+]i evoked by the Ca2+ ionophores ionomycin and A23187 caused an inhibition of outward currents as well. When the elevation of [Ca2+]i is prevented, the inhibition is decreased. These results suggest that [Ca2+]i elevation may be involved in the process. PMID- 9219621 TI - Development of somatostatin radioimmunoassay for the measurement of plasma and tissue contents of hormone. AB - A specific and sensitive radioimmunoassay was developed in our laboratory for measuring plasma and tissue somatostatin levels. The hormone content of arterial blood and skin samples of untreated and mustard oil (a specific agent causing neurogenic inflammation) treated animals was detected by this method. The somatostatin level of the inflamed tissue was significantly higher, but no difference was found between the plasma concentrations. PMID- 9219622 TI - Comparison of the effects of restacorin and flecainide on various cardiac transmembrane potassium currents. AB - The effect of restacorin (5 microM) and flecainide (5 microM) on the transmembrane ionic currents was compared in dog and rabbit ventricular muscle by applying the conventional microelectrode and patch-clamp techniques. Neither restacorin nor flecainide influenced significantly the ATP-sensitive and inward rectifier potassium currents. Flecainide moderately decreased the amplitude of the transient outward current while restacorin did not change this current. The delayed rectifier potassium current was depressed markedly by flecainide and moderately by restacorin. These results suggest that although flecainide and restacorin cause similar changes in the various transmembrane ionic currents, there are some differences between the effects of the two antiarrhythmic compounds. PMID- 9219623 TI - Effect of glutathione depletion on electrophysiological characteristics of guinea pig ventricles. AB - In this study the effect of acute and chronic glutathione depletion with L buthionine-S,R-sulphoximine (BSO) on action potential characteristics of guinea pig left ventricle papillary muscles were investigated. BSO caused significant decrease of maximum rate in rise of depolarization phase (Vmax) and duration of AP (APD) at 25%, 50%, 90% of repolarization in both cases of depletion and a slight but not significant decrease in the action potential amplitude, but did not modify the resting membrane potential. Pretreatment with bisaramil prevented the effect of BSO on APD in both cases of depletion. PMID- 9219624 TI - Influence of MCI-9042, a novel 5-HT2 receptor blocker on blood vessels of the rat. AB - We demonstrated that MCI-9042 potently inhibited the vasoconstrictory effects of serotonin in the isolated perfused hindlegs of the rat. Concentrations of MCI 9042 in the range of 0.01-1 mumol/l caused a dose-dependent inhibition of the vasoconstrictory effect of serotonin (1-30 micrograms/0.1 ml). The maximal inhibitory effect of MCI-9042 was about 97% (1 mumol/l). MCI-9042 caused a concentration-dependent shift to the right of concentration-response curve to serotonin in the rat tail artery. The present data demonstrate MCI-9042 as a potent 5-HT2 receptor antagonist. PMID- 9219625 TI - Effect of DV-7028, a novel serotonin 5-HT2 receptor antagonist on the cardiovascular system in rats. AB - The response of the cardiovascular system to DV-7028 demonstrated the complex action of this substance. In anaesthetized rats DV-7028 decreased the blood pressure and heart rate. These effects did not occur in pithed rats. The hypotensive action and bradycardia was partially reduced in vagotomized animals. DV-7028 potently inhibited the vasoconstrictory effects of serotonin in isolated perfused hindlegs of rat and caused a concentration-dependent shift to the right of response curve to serotonin in the rat tail artery. The present data demonstrate that DV-7028 is a potent 5-HT2 receptor antagonist. Besides its peripheral action, DV-7028 exerts central effects which cause hypotension and bradycardia. PMID- 9219626 TI - Influence of captopril on some haemostatic parameters in rats with ongoing process of venous thrombosis. AB - Recent laboratory findings strongly suggest that renin-angiotensin system plays an important role in regulation of haemostasis and fibrinolysis. In our previous study we showed that captopril exerts antithrombotic effect in venous thrombosis in rats. In this study we demonstrated that this effect is not a result of changes in platelet count, fibrinogen level alterations in blood coagulation pathways and fibrinolytic activity of the plasma. Further investigations are necessary to elucidate the mechanism of antithrombotic action of captopril. PMID- 9219628 TI - Non-adrenergic regulation of microcirculation evoked by antidromic stimulation of the saphenous nerve in the rat skin. AB - Electrical stimulation of the peripheral stump of the cut and perineurally capsaicin-pretreated saphenous nerve evokes antidromic vasodilatation preceded by a short vasoconstriction in the dorsal skin of the hindpaw in the rat. These microcirculatory changes were measured by laser-Doppler flowmetry. Blood flow increase induced by nerve stimulation was completely abolished by 1 microgram/kg resiniferatoxin (RTX), while the inicial blood flow decrease was significantly reduced or totally inhibited by subsequent treatments with an alpha adrenergic receptor antagonist (GYKI-12743) and a neuropeptide Y functional antagonist (alpha-trinositol) in response to 10 Hz and 3 HZ stimulations, respectively. PMID- 9219629 TI - Dose-dependent intestinal and hepatic glucuronidation and sulfatation of P nitrophenol in the rat. AB - The jejunum was able to metabolize p-nitrophenol (PNP) rapidly and to transport the metabolites efficiently back into the luminal solution. About 21, 16, 6 and 3.5% of recirculated amount of PNP could be detected in 90 minutes as glucuronide in the lumen of jejunal loop, when 20, 100, 500 or 1,000 microM PNP was perfused, which shows that the luminal appearance of p-nitrophenol-glucuronide (PNP-G) tended to saturability. Biliary excretion rate of PNP-G was lower than the luminal appearance of this metabolite, when PNP was recirculated at 20 or 100 microM concentrations, however, at higher concentrations (500, 1,000 microM) the biliary excretion exceeded the luminal appearance of this conjugate and no saturability was observed in the biliary glucuronidation of PNP. Biliary excretion of sulfate conjugate of p-nitrophenol (PNP-S) exceeded the luminal appearance of this metabolite. PMID- 9219627 TI - Effect of endothelin-1 on some hemostatic parameters in normotensive rats. AB - Some parameters of hemostasis and fibrinolysis were investigated in rats administered with endothelin-1 (ET-1). ET-1 (0.5, 1.0, 5.0 nmol/kg) dose dependently shortened the bleeding time (BT). Concomitantly significant shortening of the clotting time (CT) was observed. ET-1 produced prolongation of the activated partial thromboplastin time (APTT), whereas prothrombin time (PT) remained unchanged. ET-1 did not influence in vitro platelet aggregation induced by ADP and collagen. The euglobulin clot lysis time (ECLT) was significantly shortened after ET-1 administration. Our results suggest that ET-1 modulates the process of hemostasis and fibrinolysis in the rat. PMID- 9219630 TI - On the cardioprotective effect of adenosine. AB - In isovolumically perfused Langendorff heart preparations of guinea pigs adenosine-depending on the experimental protocol-more or less could prevent the hypoxia-induced decrease in myocardial adenosine triphosphate [ATP], creatine phosphate [CP], glycogen and increase in lactate, i.e. showed cardioprotection. PMID- 9219631 TI - Regulation of purinoceptors in guinea pig pulmonary artery: functional evidences. AB - In isolated guinea pig pulmonary arteries (precontracted with 1 microM noradrenaline) N6-cyclopentyladenosine (CPA), a selective A1 adenosine receptor agonist, exerted a concentration-dependent contraction, whereas 5'-N ethylcarboxamidoadenosine (NECA), a non-selective A1/A2 receptor agonist, in the presence of DPCPX (a highly selective A1 receptor antagonist), produced a concentration-related rapid relaxation. Pulmonary arteries obtained from guinea pigs treated with aminophylline (APH) or 8-phenyltheophylline (8-PT) for 10 consecutive days, displayed more pronounced contraction in response to CPA compared to those of solvent-treated animals. Relaxant action of NECA was, however, attenuated in arteries prepared from methylxanthine-treated guinea pigs. Opposite changes were found in vascular tissues excised from chronically dipyridamole(DP)-treated guinea pigs. PMID- 9219632 TI - Prolonged treatment with antidepressants increases the 5-HT1A-mediated inhibition of hippocampal neurons without changing the 5-HT1A receptor binding. AB - The effect of repeated treatment with antidepressant drugs imipramine, (+)oxaprotiline and paroxetine on neuronal responsiveness to 5-HT and the 5-HT1A receptor agonist 8-OH-DPAT was examined in the hippocampal slice preparation from the rat. 5-HT and 8-OH-DPAT decreased the amplitude of population spikes evoked in the CA1 cell layer by electrical stimulation of the stratum radiatum. The antidepressant drugs, administered for 2 weeks, produced a significant increase in the inhibitory effect of 5-HT and 8-OH-DPAT. Repeated treatment with imipramine did not change the density of 5-HT1A receptors in the hippocampus suggesting that the increase in 5-HT1A responsivity may not involve an increase in the receptor density. PMID- 9219633 TI - The effect of chronic diazepam treatment on hypoxia-induced alterations in functional activity of guinea pig myocardium. AB - The concentration-related sensitization of guinea pig left atrium to adenosine in the presence of diazepam is well established. It was found in our experiments that the cardiodepressive action of hypoxia is significantly enhanced by diazepam in the left atrial myocardium. In atrial preparations obtained from guinea pigs treated with diazepam for 10 days, the hypoxia-induced depression of myocardial contractility was not altered. These results indicate that diazepam-treatment does not impaire the hypoxic tolerance of myocardium. PMID- 9219634 TI - The role of NMDA receptors in central action of angiotensin II. AB - The influence of noncompetitive (MK-801), competitive (AP-7) and the antagonist of polyamines site of NMDA receptor (arcaine) on the central activity of angiotensin II (A II) was studied. The open field test, conditioning of active avoidance responses (CARs) and passive avoidance situation was used to investigate learning and memory in rats. All used antagonists decreased beneficial action of A II on these processes. PMID- 9219636 TI - Intra- and extracellular isoforms of PR-3 class chitinase in virus-infected cucumber plants. AB - Cucumber (Cucumis sativus L. cv. Laura) contains three different isoforms of chitinase (EC 3.2.1.14), thought to be involved in the defense against a pathogen. Using a highly specific rabbit antiserum raised against a predominant, extracellularly localized, virus-inducible acidic chitinase (p28), two additional enzyme isoforms with differential mode of compartmentalization were identified. Immunoblot analysis of the fractionated plant extracts separated by denaturing and native (anodic and cathodic) polyacrylamide gel electrophoresis (PAGE) revealed that the chitinase p25 (M(r) 25.5 K), is a basic, extracellular isoform and the chitinase p24 (M(r) 24.6 K) is an acidic, probably intracellular isoform of the enzyme. PMID- 9219635 TI - Combined protective effect of an immunostimulatory bacterial preparation and rimantadine in experimental influenza A virus infection. AB - The protective effect of an immunostimulatory bacterial preparation, cytoplasmic membranes of Escherichia coli WF stable protoplast type L-forms (CM) alone and in combination with the selective antiviral drug rimantadine was evaluated in experimental influenza A/Aichi/2/68 (H3N2) virus infection in mice. In sublethal infection, CM administered intraperitoneally (i.p.) 7 days before virus exposure in a single dose of 25 mg/kg did not reduce significantly the virus lung titers. In lethal infection, CM applied in the same way weakly reduced the mortality rate. The combined application of CM with rimantadine resulted in synergistically increased protection, determined on the basis of virus lung titers, lung consolidation, mortality rates, protective indices, and survival times. PMID- 9219637 TI - High concentration of recombinant murine interferon-beta enhances the growth of Orientia (formerly Rickettsia) tsutsugamushi Gilliam in mouse L929 cells. AB - We studied the effect of recombinant murine interferons (rMuIFNs) on the growth of Orientia (formerly Rickettsia) tsutsugamushi Gilliam in mouse L929 cells. Rickettsial growth was measured by flow cytometry. rMUIFN-gamma inhibited the growth of O. tsutsugamushi at the concentrations of 100 i.u./ml and 1,000 i.u./ml in accord with previous reports. Relatively low concentrations (10 i.u./ml and 100 i.u./ml) of rMUIFN-beta also inhibited the growth of O. tsutsugamushi. On the other hand, high concentrations (1,000 i.u./ml and 10,000 i.u./ml) of rMuIFN-beta enhanced the growth of the rickettsia. This enhancement of rickettsial growth was blocked by anti-murine IFN-beta monoclonal antibody (MoAb). rMuIFN-beta also enhanced the growth of Rickettsia sibirica 246 in L929 cells to some extent. PMID- 9219638 TI - Effects of brefeldin A on the expression and transport of influenza A virus haemagglutinin, M1 and M2 proteins within the cell. AB - Brefeldin A (BFA) decreased the expression of influenza A virus haemagglutinin (HA) and M2 protein on the plasma membrane of virus-infected MDCK cells. It caused a retention of M1 protein in the cell nucleus and a decrease of its expression on the plasma membrane. On the other hand, an increased labelling of the cytoplasmic domain of M2 protein on the plasma membrane in BFA-treated cells was observed in contrast to the labelling in BFA-untreated cells. The effects of BFA on the microtubules and cellular motors are discussed. PMID- 9219639 TI - Loss of thymidine kinase activity due to a base deletion in a candidate vaccine strain of bovine herpesvirus 2. AB - The nucleotide (nt) sequence of the thymidine kinase (TK) gene (a 918 nt long coding region) of two TK-deficient (TK) strains of bovine herpesvirus 2 (BHV-2) was determined. The candidate vaccine strain C290BU5, which was no longer able to cause disease, was found to have an A deletion after nt 887 of the TK gene with a predicted change of His 296 to Pro, altering the last 10 amino acids (aa) and extending the gene by another 34 aa. The strain which still caused disease, C290BU3, had a T insertion after nt 16 causing a predicted chain termination after only 16 aa. PMID- 9219640 TI - Summing up, weak immunomodulating signals of Tahyna virus and immunomodulating drugs induce immunosuppression in mice. AB - The successive injection of non-immunomodulating doses of Tahyna virus (100 LD50) and non-immunomodulating doses of immunomodulating drugs, such as purified staphylococcal toxoid or glucosaminylmuramyldipeptide (Likopid), to mice were accompanied by a decrease in the IgM plaque-forming cell response to sheep red blood cells. PMID- 9219641 TI - Screening of apple mosaic virus in hop cultivars in the Czech Republic by reverse transcription-polymerase chain reaction. AB - Thirteen cultivars of hop (Humulus lupulus L.) were tested by reverse transcription-polymerase chain reaction (RT-PCR) for the presence of apple mosaic virus (ApMV). The virus was detected in various amounts in all tested cultivars. Control hop clones derived from tissue cultures, treated by thermotherapy and maintained in greenhouse were virus-free. The procedure for sample preparation and RT-PCR of ApMV is described in detail. PMID- 9219642 TI - Stable transfection of provirus of human immunodeficiency virus into a murine packaging cell line. AB - In order to generate HIV (murine leukemia virus (MuLV)) pseudotypes, HIV genome was transfected into the ecotropic murine packaging cell line (GP+E86) and four of the nine transfected clones were extensively characterized. One clone (801), harbouring a full copy of integrated HIV sequences, exhibited a detectable level of intracellular HIV p24 antigen expression. Northern blot analysis revealed that clone 801 expressed all three classes of HIV mRNAs. Multispliced 2 kb mRNAs were detected in another clone (8.14). Two other clones (1.31 and 1.32) also exhibited a complete HIV provirus, but did not show any viral expression, as evaluated by Northern blot analysis or HIV p24 ELISA. Reverse transcription-polymerase chain reaction (RT-PCR) experiments revealed the presence of full length genomic RNA in four transfected clones, which were extensively characterized. A co-cultivation of clone 801 with human CD4' cells resulted in syncytia formation. By electron microscopy, mature HIV particles were observed after co-cultivation of uninfected C8166 cells with 801 cells. These results demonstrated that the murine clone was stably transfected with the complete HIV genome and was capable of shuttling infectious HIV to human cells. Clone 801 was co-cultivated with murine NIH-3T3 fibroblasts. In several experiments, HIV infection of NIH-3T3 cells was revealed by PCR technique. Thus, 801 cells appear to produce low levels of HIV (MuLV) pseudotypes capable of transferring the HIV genome into mouse cells. PMID- 9219643 TI - Detection and identification of the Newcastle disease virus infection by electron and immunoelectron microscopy. AB - Various electron microscopic (EM) and immunoelectron microscopic (IEM) techniques were used to demonstrate and identify the Newcastle disease virus (NDV) infection. By IEM, the number of virions in native allantoic fluids was increased 50-100 times in comparison with direct EM. The immunogold staining showed that a number of immunogold particles were specifically bound to the antigen determinants located on the virion surface and these results were much easier to interpret. The obtained results showed that the EM and IEM can be successfully employed for a precise and rapid detection of NDV as well as for identification of this infection among other viral or bacterial infections. PMID- 9219644 TI - Biology of tick-borne encephalitis virus. AB - Tick-borne encephalitis (TBE) virus is an important human pathogen belonging to the genus Flavivirus within the family Flaviviridae. The genome of the TBE virus is a single-stranded RNA (ssRNA) molecule of positive polarity encoding all the viral proteins within a single open reading frame (ORF). TBE virus shares common physical and genetic characteristic of the flavivirus genus. Two subtypes of the TBE virus have been described: (1) European, endemic in many parts of Europe and transmitted by Ixodes ricinus ticks, and (2) Far Eastern (Russian spring summer encephalitis (RSSE) virus), endemic in Far East and transmitted by Ixodes persulcatus ticks. PMID- 9219645 TI - Unacceptable "occupational" exposure to toxic agents among children in Ecuador. AB - To document the problem of child labor as a health issue, we report here three case-studies in Ecuador: exposure to mercury among gold washers, exposure to organophosphates and carbamates in the fruit-growing industry, and exposure to solvents among shoe cleaners. We measured the relevant biological indicators of exposure (mercury in urine, urinary levels of phenols, and acetylcholine esterase in erythrocytes) among selected samples of 10 children for each working place. In all the case studies, the values of the biological indicators showed elevated exposure to well-known toxicants, which are now rare in developed countries, even among adult workers. The findings meld with a previously reported case study of intoxication from inorganic lead among children employed in the manufacture of roof tiles in Ecuador. This study highlights the need to properly evaluate and control the potential health effects due to exposure to toxic substances among children employed in different occupations in several parts of the world. PMID- 9219646 TI - Implementing participatory ergonomics teams among health care workers. AB - Three participatory ergonomics teams have been established among healthcare workers in a metropolitan medical center. Three teams, consisting of orderlies, intensive care unit nurses, and laboratory workers, were selected to provide a diversity of work activities and educational backgrounds. The effectiveness of these teams was assessed by observations of team interactions, by team members' perceptions of their effectiveness, and by the teams' success in identifying problems and implementing solutions. After 1 year, one of the three groups has been highly effective by these measures. To varying degrees, the groups encountered competing time demands and obstacles in implementing solutions within current administrative structures. For some groups of health care workers, participatory ergonomics teams seem to be an effective strategy to improve health and safety. This approach may not be feasible in all areas of health care, especially in high-demand clinical areas where patient needs may take precedence over the safety of health care workers. PMID- 9219647 TI - Postal questionnaire study of disability associated with latex allergy among health care workers in Finland. AB - In this study, the association between natural rubber latex (NRL) sensitization and work ability index (WAI) among health care workers was investigated. Furthermore, the diagnostic sensitivity and specificity of a postal questionnaire as a screening device of NRL allergy was evaluated. The study population consisted of 32 female health care workers with an occupational latex allergy, and 51 control subjects who were individually matched for age and occupation. A self-administered two-part questionnaire, including seven items of a work ability index (WAI), as well as questions on glove-related symptoms, was mailed to the subjects. The median age for NRL allergic subjects was 40 years (range 23-62), and the diagnosis of occupational latex allergy had been made six years (range 2 16) before the present study. The WAI scores were on average lower among the sensitized subjects as compared with their nonsensitized controls. Even after removing the contribution of the presence of allergic eczema, diagnosed by a physician, from the original WAI score, the proportion of NRL allergic subjects and the control subjects in the good work ability category were 34% and 53%, respectively. Ten health care workers (31%) had changed occupation and one early retirement had occurred after sensitization to NRL. The sensitivity and specificity of the present self-administered questionnaire as an indicator for latex allergy was 84% and 98%, respectively. In conclusion, there is a clear association between NRL allergy and a decrease in the WAI among health care workers, which cannot be explained by age, gender, profession, or history of atopy. PMID- 9219648 TI - Sinonasal cancer and occupational exposure to textile dust. AB - Data from a case-control study conducted at 27 hospitals in France in 1986-88 were analyzed to examine the association between exposure to textile dust and sinonasal cancer. The study included 207 cases and 409 controls. Detailed information on occupational history and other potential risk factors for sinonasal cancer was collected. Exposure to textile dust (probability and level of exposure, type of textile fiber) was assessed by an expert in industrial hygiene. Among women, exposure to textile dust was associated with an elevated risk of squamous cell carcinoma (odds ratio (OR) = 2.45, 95% confidence interval (CI) = 0.85-7.06, nine exposed cases) and adenocarcinoma (OR = 3.70, 95% CI = 0.56-24.4, three exposed cases). For squamous cell carcinomas, the risk increased with the duration and the level of exposure (P < 0.05): the ORs for the low, medium, and high level of cumulative exposure were 1.00 (95% CI = 0.10-9.43), 2.43 (95% CI = 0.54-11.1), and 3.57 (95% CI = 0.92-13.8), respectively. There was also a limited evidence of an excess risk of squamous cell carcinomas among men exposed to high levels of textile dust (OR = 2.18, 95% CI = 0.65-7.30, four exposed cases). Because of the strong association between wood-dust exposure and adenocarcinoma, an independent effect of textile dust on this type of cancer could not be studied among men. The risks associated with the different types of textile fibers (cotton, wool, and synthetic fibers) were similar and the results did not permit to incriminate a particular type of textile. PMID- 9219649 TI - Acute arsine intoxication as a consequence of metal burnishing operations. AB - The report concerns a 30-year-old factory worker, employed in a small galvanizing plant for over ten years in the burnishing, copper- and nickel-plating of small metal articles for the shoe industry. Acute arsine poisoning was attributed to the use of a dilute solution of CuSO4 (3%), HCl (32%), and As2O3 (2%) for burnishing metal (Fe-Zn) shoelace eyelet holes, in the absence of local exhaust ventilation and with no respiratory protection. Arsine caused severe intravascular hemolysis with a rapid drop in hematocrit and hemoglobin levels. Other body organs were involved as a result of the hypoxic effect of anemia and hemolysis, or as a direct toxic effect of the arsine itself. Our experience confirms that exchange transfusion is capable of rapidly arresting the adverse effects of arsine. The importance of preventive measures and worker information to avoid acute arsine poisoning is emphasized. PMID- 9219650 TI - Endocrine profiles of male workers with exposure to trichloroethylene. AB - The objective of this study was to examine the endocrine profiles of a group of male workers chronically exposed to trichloroethylene (TCE) in an electronics factory. A total of 124 workers participated in a preliminary study, for which 85 satisfied the selection criteria and were recruited to take part in a more detailed study. Each of the 85 workers had urine collected and analyzed for trichloroacetic acids (TCA) on the day blood was taken for analysis of serum testosterone (T), follicle-stimulating hormone (FSH), dehydroepiandrosterone sulphate (DHEAS), and sex-hormone binding globulin (SHBG). Environmental TCE exposures were conducted for 12 workers. The geometric mean concentration of environmental TCE was 29.6 ppm (range 9-131) and the mean urine TCA was 22.4 mg/g creatinine (range 0.8-136.4). The results showed that years of exposure to TCE were significantly correlated with DHEAS and negatively correlated with SHBG and T levels. Serum FSH, T, and SHBG levels showed a gradual decline with increasing years of exposure to TCE. This dose-response decrease indicated that there was a disruption of peripheral endocrine function. This disruption could be a result of TCE-induced liver malfunction. The most dramatic change was that the increase in DHEAS concentration was associated with years of exposure to TCE, rising from 255 to 717.8 ng/ml for < 3 to > or = 7 years exposure, respectively. This evidence suggests that chronic exposure to TCE may also affect adrenal function. These findings, however, must be confirmed by further investigations. PMID- 9219651 TI - A cohort mortality study of foundry workers. AB - Since the 1970s, hygienic improvements have led to a reduction in the level of airborne pollutants in Danish foundries. This mortality study reflects the exposure situation prior to 1970, and the findings may be used as a baseline for future evaluations of the preventive impact of reduced exposure. Mortality data were derived from a historical cohort study in which 3,056 foundry workers were compared with 43,024 workers employed in other industries. The foundry workers' life-long risk of dying from pneumoconioses averaged 2% and the corresponding standardized mortality ratio (SMR) equaled 7,368 (95% confidence interval (95% CI): 4,029-12,363). Excess mortality was also seen for chronic bronchitis and emphysema (SMR = 132, 95% CI: 98-185). Nonsignificant increases were seen for buccal cancer, stomach cancer, colon cancer, and urothelial cancer. In conclusion, Danish foundry workers exposed prior to 1970 seem to suffer an excess risk of devastating lung disease of occupational origin. PMID- 9219652 TI - A cohort study of Swedish capacitor manufacturing workers exposed to polychlorinated biphenyls (PCBs). AB - Mortality and cancer incidence were investigated among 242 male capacitor manufacturing workers, exposed to polychlorinated biphenyls (PCBs) for at least six months between 1965 and 1978. Mortality and cancer incidence were followed from 1965 to 1991. There was a significantly increased mortality from cardiovascular diseases among those employed for at least five years in high exposed jobs, with a latency of 20 years. There were two cases of cancer of the liver and bile ducts, which previously has been associated with PCB exposure, both in epidemiological and animal experimental studies. No data on smoking habits were available. The study supports some previous findings of an increased risk of cancer of the liver and bile ducts after exposure to PCBs. The reason for the excess of cardiovascular deaths in the high-exposure group is not known and deserves evaluation in future studies. PMID- 9219653 TI - Mortality studies of machining fluid exposure in the automobile industry. V: A case-control study of pancreatic cancer. AB - Results are presented from a case-control study of 97 cases of pancreatic cancer nested in a cohort of workers from three automobile manufacturing plants. Risk was examined for lifetime exposure to straight, soluble, and synthetic metalworking fluids, as used in specific machining or grinding operations, as well as for constituents of the fluids. Pancreatic cancer was associated with exposure to synthetic fluids in grinding operations, with an odds ratio of 3.0 (95% CI: 1.2-7.5) among those with more than 1.4 mg/m3-years of exposure. We were unable to examine synthetic exposure in the absence of grinding because there was virtually no exposure to synthetics in machining operations in this study population. Although a disproportionately high percent of the cases were black, no black workers had any exposure to synthetic fluids, and no other measured exposure was found to be related to risk. Thus, the previously documented excess risk of pancreatic cancer among blacks in this cohort remains unexplained. PMID- 9219654 TI - Liver cancer among employees in Denmark. AB - To test the hypothesis that occupational exposure to chemical agents-particularly organic solvents in certain industries-may cause primary liver cancer (PLC), a nested case-control study of PLC cases from the Danish Cancer Registry and an age and sex-stratified random sample of controls from the Central Population Register in Denmark were linked with files of a national supplementary pension fund. Employment histories since April 1964 were obtained for 973 cases histologically classified as hepatocellular carcinoma, cholangiocarcinoma, or combined hepatocellular and cholangiocarcinoma and 15,348 controls. Men from 35 different industrial branches, women from 7 branches, and both men and women from 3 branches had an excess risk of PLC, with an odds ratio of (OR) > 1.0; 29 branches had an OR of liver cancer in excess of 3.0. Women from bookprinting and offset printing industries had an OR above 10. Only male farmers had an OR below unity (0.41). Employees from breweries, restaurants, hotels, motels, and distilleries had an increased OR of both PLC and esophageal cancer. PMID- 9219655 TI - Liver function alterations in synthetic leather workers exposed to dimethylformamide. AB - A cross-sectional study of the prevalence of chronic liver function alterations was performed in 75 workers employed in a synthetic leather factory, exposed to dimethylformamide (DMF) air concentrations below threshold limit values (30 mg/m3). Biological monitoring among workers revealed acceptable urine levels of monomethylformamide (NMF) on average, but the very wide range indicated that occasional overexposure was possible. The worker survey showed a high percentage of disulfiram-like symptoms (50%) and liver function abnormalities (22.7%), compared with a demographically similar group of unexposed workers. Covariance analysis (ANCOVA) revealed that enzyme levels were significantly higher in exposed workers than in controls after data were corrected for age, alcohol consumption, body mass index, and cholesterol levels. The authors conclude that DMF can cause liver diseases even if air TLVs are respected, because accidental contact with liquid DMF can significantly increase DMF uptake. In this situation, air monitoring is no longer sufficient to evaluate worker exposure. PMID- 9219656 TI - Follow-up of biologic monitoring results in cadmium workers removed from exposure. AB - Biological monitoring and exposure monitoring data for employees at a nickel cadmium battery production facility were made available to OSHA for review. Sixteen employees were medically removed from occupational exposures to cadmium due to elevated levels of biological parameters (CdB, Cdu, B2U). While the biological monitoring parameters for most workers significantly declined during the 18 months of medical removal, the biological parameters for only one employee's values returned to the normal range. Only one worker had frank renal dysfunction, based on beta-2-microglobulin levels at the time of removal; this dysfunction remained throughout the 17 months of observation after medical removal. Significant policy implications of medical removal protection beyond the current 18-month period provided by the cadmium standards exist and require physician discretion. Mitigating issues which may make it ethically appropriate to return an employee to work despite elevated biologic monitoring parameters are also discussed. PMID- 9219657 TI - Respiratory symptoms and spirometry in experienced coal miners: effects of both distant and recent coal mine dust exposures. AB - The goal of this study was to determine whether respiratory symptoms were associated with the lower concentrations of respirable coal mine dust that were required by the U.S. Coal Mine Health and Safety Act (CMHSA) of 1969. The subjects were 1,866 male miners who had participated in the National Study of Coal Workers' Pneumoconiosis (NSCWP) and been tested at least twice, initially in either Round 1 (R1) (1969-71) or Round 2 (R2) (1972-75) and then finally in Round 4 (R4) (1985-88). Self-reported information elicited with a standardized questionnaire was used to determine the presence at the final round (i.e., R4) of chronic bronchitis, shortness of breath, and wheeze. Cumulative coal mine dust exposure was characterized for both the pre- and post-CMHSA periods. Controlling for age and other potential confounders, increased risks for the symptoms were associated with higher levels of both measurements of exposure. Moreover, the adverse effects of the lower, post-CMHSA exposure were evident for shortness of breath and wheeze especially among subjects who had little pre-CMHSA coal mining experience. These findings provide additional evidence of the limitations of the current 2.0 mg/m3 coal mine dust standard to prevent respiratory disease. PMID- 9219658 TI - Prevalence of physician-diagnosed asthma by occupational groupings in Manitoba, Canada. AB - The objective of this research was to determine whether there are differences in the rate of physician-diagnosed asthma in various occupational groups. A prevalence survey using a population-based administrative database of a sample of the labor force in Manitoba, Canada was used. A sample of 22,561 individuals who were in the labor force at the time of the 1986 census were linked to the provincial administrative health database. The frequency of physician-diagnosed asthma and other obstructive respiratory conditions were measured. A case of asthma was defined as having at least three physician contacts for asthma between April 1, 1986, and March 31, 1990. Data on potential confounding factors such as age, gender, area of residence, income, and education were also available. The results showed that frequency of physician-diagnosed asthma by occupational grouping ranged from a low of 0.1/100 workers to a high of 4.8/100 workers. Three occupational groups, 1) other teaching and related occupations (SOC 279) (OR 2.54, 95% CI 1.18-5.44); 2) fabricating, installing, and repairing occupations of electrical electronic and related equipment (SOC 853) (OR 2.37, 95% CI 1.05 5.33); and 3) other occupations in laboring and other elemental work (SOC 992) (OR 2.51, 95% CI 1.21-5.24) were found to have elevated odds ratios for physician diagnosed asthma. Datasets linking occupation and health care utilization may be useful tools for surveillance of work-related diseases in general, and for asthma in particular. However, further work should be done utilizing larger databases to determine the overall usefulness of this approach. PMID- 9219659 TI - Paternal occupational exposure around conception and spina bifida in offspring. AB - A multi-center case-referent study was conducted on the relation between paternal occupational exposure and spina bifida in offspring. Cases were born between 1980 and 1992 in The Netherlands. Referents were recruited from hospitals and from the general population. Postal questionnaires were used to gather information on occupation and potential confounders. Through job-specific telephone interviews with 122 case fathers and 411 referent fathers, detailed exposure information was collected on specific tasks, the use of chemical or physical agents, frequency of exposure, and use of protective equipment. The study yielded statistically significant associations between spina bifida and low exposure to welding fumes (OR = 1.6, 95% CI: 1.0-2.6) and low exposure to UV radiation during welding (OR = 2.6, 95% CI: 1.2-5.6), and suggestive findings of an association between spina bifida and moderate or high exposure to cleaning agents, moderate or high pesticide exposure (OR = 1.7, 95% CI: 0.7-4.0), and stainless steel dust (OR = 2.0, 95% CI: 0.8-5.2). No associations were identified for other paternal occupational exposures, such as organic solvents. PMID- 9219660 TI - Musicians' playing-related musculoskeletal disorders: an examination of risk factors. AB - Several studies have shown that playing-related musculoskeletal disorders (PRMDs) present a significant health problem for musicians. To examine physiological, psychological, and behavioral risk factors of musicians' PRMDs, data for a case control analysis were collected from classically-trained professional and university student musicians in the Canadian province of Ontario in 1994. Two hundred and eighty-one subjects completed a self-report questionnaire and hypermobility and hand-span measurements. Cases were identified according to an operational PRMD definition developed by musicians and health care professionals in a qualitative study. Logistic regression was used to compare data from 44 prevalent PRMD cases who had no previous history of a PRMD, and 90 controls who had never experienced a PRMD. Data from all subjects were analyzed to examine the role of a prior PRMD on the risk of a current PRMD. This study suggests that females and string players were at a higher PRMD risk. A number of other individual characteristics were also important determinants of the development of a PRMD. Warming up before and taking breaks during practice sessions protected the subject from a PRMD. Given the high proportion of musicians who experience PRMDs, prevention programs are warranted. PMID- 9219661 TI - Cancer feasibility studies among migrant farmworkers. The Farmworker Epidemiology Research Group. PMID- 9219662 TI - Women, work, and health. AB - The U.S. Bureau of National Affairs has conducted several surveys asking women to rate the seriousness of 11 hazards thought to affect female workers. In 1995 the women respondents ranked them in the following order: 1) stress, 2) repetitive motions, 3) AIDS, 4) violence, 5) VDTs, 6) indoor air pollution, 7) hepatitis, 8) injury on the job, 9) reproductive hazards, 10) tuberculosis, and 11) other infectious diseases. A parallel list of 11 hazards thought to affect male workers would look very different. The purpose of this paper is to explore why this is so and what it implies for the occupational health research agenda. PMID- 9219663 TI - A battle for compensation for Welsh coal miners: JS Haldane v "Sericite" Jones, 1932-1934. AB - Toward the end of the 1920s, technical advances in mining led to an increase in airborne burdens of dust in the South Wales coal mines. This coincided with a dramatic increase in the incidence of disability and death from respiratory disease among the miners. For their condition to be compensable, claimants were required to have worked with rock containing more than 50% 'free silica.' Dr W.R. Jones, a mining geologist, was asked to help obtain compensation for those claimants who could not satisfy the 'free silica' condition. He was unable to identify high-silica rocks where none had been said to exist. He did however, successfully argue the brief against the eminent Professor J.S. Haldane (who was the dominant authority, having had lengthy experience in the field of health and mining), for the fibrous form of sericite being commonly the important agent responsible for pneumoconiosis. As a consequence, the category of miner eligible for compensation was broadened. Evidence was gathered worldwide that supported the hypothesis that silicates and not just crystalline silica could cause pneumoconiosis. Despite the suspicions raised about the special power of mineral fibers during this public debate, some 40 years were to elapse before potential health hazards from fibers other than asbestos were to be taken seriously and investigated. PMID- 9219664 TI - Paternal lead exposure, offspring birth weight, and sex ratio. PMID- 9219665 TI - Partition of circulating lead between plasma and red cells does not seem to be different for internal and external sources of lead. PMID- 9219666 TI - Serum (plasma) lead, blood lead, and bone lead. PMID- 9219667 TI - Re: Predictors of return to work after carpal tunnel release. PMID- 9219668 TI - A numerical model of nasal odorant transport for the analysis of human olfaction. AB - The transport and uptake of inspired odorant molecules in the human nasal cavity were determined using an anatomically correct three-dimensional finite element model. The steady-state equations of motion and continuity were first solved to determine laminar flow patterns of odorous air at quiet breathing flow rates. The air stream entering the ventral tip of the naris traveled to the olfactory slit, and then passed through the slit in nearly a straight path without forming separated recirculating zones. The fraction of volumetric flow passing through the olfactory airway was about 10%, and remained nearly constant with variation in flow rate. The three-dimensional inspiratory velocity field was used in the solution of the uncoupled steady convective-diffusion equation to determine the concentration field in the airways and odorant mass flux at the nasal walls. The mass-transfer boundary condition used at the nasal cavity wall included the effects of solubility and diffusivity of odorants in the mucosal lining, and the thickness of the mucus layer. The total olfactory flux of odorants, that is highly correlated with perceived odor intensity, was determined as a function of all transport parameters in our model. Increase in nasal flow rate at a constant inlet concentration resulted in an increase in total olfactory uptake for all odorants. However, with increase in flow rate, the fractional uptake, i.e., total olfactory flux normalized by convective flux at the inlet, decreased for poorly soluble odorants, while it increased for highly soluble odorants. The pattern of flux (or imposed patterning) across the olfactory mucosa, that carries information concerning odor identity, was also determined as a function of transport parameters. There was an overall decrease in odorant flux as the location on the olfactory surface was varied from the anterior towards the posterior and from the inferior towards the superior ends. The flux pattern became more uniform, i.e., the steepness of the flux gradients across the olfactory surface decreased, as the mucus solubility of the odorants decreased. Different odorants generated discernibly different flux patterns across the olfactory mucosa that may contribute to the encoding of odor quality. Variation of total olfactory flux with time after cessation of airflow was determined by solving the unsteady diffusion equation in the air-phase. The flux decreased approximately exponentially with time. The rate of decay decreased as solubility and diffusivity decreased, but was very rapid over a wide range of the parameters, with time constants of less than 0.5 s for most odorants, implying a rapid decrease in perceived odor intensity with cessation of nasal airflow. PMID- 9219669 TI - Signal-induced Ca2+ oscillations through the regulation of the inositol 1,4,5 trisphosphate-gated Ca2+ channel: an allosteric model. AB - We propose a molecular model for InsP3-sensitive Ca2+ oscillations based on the allosteric properties of the InsP3 receptor/Ca2+ channel. Our model interprets the cooperatively towards InsP3 saturation, of calcium efflux from intravesicular stores as well as the absence of cooperativity in the binding process of InsP3 on the receptor. It takes into account quantitatively the two antagonist, concentration-dependent effects (fast activator and slow inhibitor) that cytosolic Ca2+ exerts on the InsP3 receptor/Ca2+ channel. Assuming that a single pool of releasable Ca2+ exists in the endoplasmic reticulum, the model leads to cytosolic and intravesicular oscillations in Ca2+ at fixed InsP3 concentration. Activation of the receptor by cytosolic calcium is essential for the triggering of oscillations whereas the slow Ca2+ inhibition effect is irrelevant in this respect, although this regulation loop might prevent the system from entering the unstable domain in absence of a true agonist stimulation. Activating cytosolic Ca2+ and InsP3 have quite distinct functions for the induction of Ca2+ release: cytosolic Ca2+ triggers oscillations whereas InsP3 only brings the receptor into a potentially oscillatory regime. Hence, the increasing slope of Ca2+ spiking is constitutively independent from the intensity of the hormonal stimuli in our model, in accord with experimental observations. Comparisons with other existing models are given and additional possible coupling mechanisms are discussed in order to explain particular facts (such as possible oscillations of InsP3) which do not depend on the intrinsic properties of the oscillator. PMID- 9219670 TI - Mutation rates as adaptations. AB - In order to better understand life, it is helpful to look beyond the envelop of life as we know it. A simple model of coevolution was implemented with the addition of a gene for the mutation rate of the individual. This allowed the mutation rate itself to evolve in a lineage. The model shows that when the individuals interact in a sort of zero-sum game, the lineages maintain relatively high mutation rates. However, when individuals engage in interactions that have greater consequences for one individual in the interaction than the other, lineages tend to evolve relatively low mutation rates. This model suggests that one possible cause for differential mutation rates across genes may be the coevolutionary pressure of the various forms of interactions with other genes. PMID- 9219671 TI - A dynamic model for the morphogenesis of the late vertebrate lens. AB - A mathematical model is presented for the morphogenesis of the post-vesicular vertebrate lens with an umbilical suture. The lens is modeled as having four compartments: anterior epithelium (germinative and central anterior zones), recruitment zone (transitional zone), cortex (discrete concentric cohorts of secondary cortex fiber cells, each cohort treated individually), and nucleus. Equations are written to describe the time evolution of the cohorts; their shapes collectively determine the shape of the lens. The growth of cell volume is exponential, with different rates in the cortex and epithelium; recruitment of epithelium cells into the cortex is described as resulting from an overproduction of epithelial basal (capsular) surface in the anterior epithelium. The equations contain three dimensionless numbers determined by the physiology of the epithelium and cortex cells. Solutions are stable attractors in a morphological space. All solutions entail exponential growth of the lens diameter; a portion of parameter space corresponds to exponential growth superimposed on large amplitude oscillations in lens shape. Emergent time-scales for increase in lens size and oscillation period are an order of magnitude longer than the cellular growth time scales. The lens shapes tend to a family of stable scaling solutions, the shapes of which remain unchanged as the lens grows. The model is applied to morphological data for the chick and lamprey lenses. The dynamics described are seen as exemplifying an auto-regulatory morphogenesis process wherein a system passes through a sequence of developmental stages. Each stage is characterized by its own fixed informing geometry (a set of defining spatial relationships), within which a growth process unfolds autonomously, generating a dynamically stable structure. The developing system invokes a means of forgetting dated structural information; this dissipation is necessary to the pattern formation process. PMID- 9219672 TI - The axon as a metabolic compartment: protein degradation, transport, and maximum length of an axon. AB - We present a model that predicts the maximum axonal length from the apparent velocity of slow axonal transport and cytoskeletal protein half-life. The model assumes that in mature axons the apparent velocity of slow transport varies with position, but that the density of cytoskeletal proteins and protein degradation are uniform. The model predicts that the apparent transport velocity of cytoskeletal proteins if highest near the cell body and decreases linearly along the axon, and that when axons branch the apparent velocity of transport decreases across the branch point. The predictions of this model are shown to be consistent with experiments. These results explain the variation in these fundamental metabolic parameters in different axons and species. PMID- 9219673 TI - A guide to ophthalmic pharmacy medicines in the United Kingdom. PMID- 9219674 TI - Gonioscopy: evaluation of the anterior chamber angle. Part II. AB - Gonioscopy is the standard procedure for examination of the anterior chamber angle, and is an invaluable technique in primary eye care. The clinician must practise the technique and observe many angles, because enormous but often subtle variation can be seen by the experienced observer in both normal and abnormal angles. Gonioscopy is essential to master, not only for the assessment of patients' risk for angle closure following dilation, but also in the diagnosis and subsequent management of the acute and chronic glaucomas and many other anterior segment disorders. PMID- 9219675 TI - Optical management of monocular pseudophakia in a child. PMID- 9219676 TI - Corneal sensitivity in health and disease. PMID- 9219677 TI - Calcium phosphate particle induction of metalloproteinase and mitogenesis: effect of particle sizes. AB - Calcium pyrophosphate dihydrate (CPPD) and basic calcium phosphate (BCP) crystals [hydroxyapatite (HA), octacalcium phosphate, tricalcium phosphate] are common in osteoarthritis knee effusions, and are often associated with low-grade synovial proliferation and inflammation. Calcium-containing crystals including HA, are known to have a number of biologic effects on culture cells such induction of mitogenesis, stimulation of Prostaglandin E2 (PGE2) production via the phospholipase A2/cyclo-oxygenase pathway, activation of phospholipase C and inositol phospholipid hydrolysis, induction of metalloproteinase synthesis and induction of proto-oncogenes (c-fos and c-myc). While endocytosis of HA particles is prerequisite of the mitogenic effect of calcium-containing crystals in fibroblasts, it is not known whether endocytosis is required for crystal-induced metalloproteinase synthesis. In the present series of experiments, we examine the effect of three different sizes (106, 46, and 17 microns mean diameters) well characterized spherical HA particles on the induction of mitogenesis and metalloproteinase synthesis on human fibroblasts. We showed that endocytosis is required for HA particles to induce synthesis of metalloproteinases. PMID- 9219678 TI - Arthroscopic evaluation of potential structure modifying activity of hyaluronan (Hyalgan) in osteoarthritis of the knee. AB - OBJECTIVE: Several reported studies suggest that repeated intra-articular injections of hyaluronan result in sustained relief from pain and functional disability in patients with knee osteoarthritis. Several in vivo data suggest that hyaluronan might have a beneficial structural effect in osteoarthritis. The objective of the study was to evaluate the potential structure-modifying effects of Hyalgan (500-730 kDa molecular weight), a highly-purified sodium hyaluronate. DESIGN: Patients with painful knee osteoarthritis (ACR criteria) were enrolled in a prospective, controlled study of 1-year duration. After randomization, either conventional therapy or three cycles (every 3 months) of three intra-articular injections of Hyalgan (once a week during 2 weeks) were given. Clinical outcome was added using pain visual analog score (VAS), functional impairment: Lequesne's index, quality of life: arthritis impact measurement scale (AIMS2) and structural outcome using X-rays: joint space narrowing and arthroscopy: global assessment using VAS, SFA scoring and grading systems. RESULTS: Of the 39 recruited patients, 36 completed the 1-year trial (19 in the Hyalgan group and 17 in the control group). There was no difference between groups at entry. Between-group comparison for changes in clinical parameters reached statistical significance for the quality of life index (AIMS2: -0.4 +/- 0.7 vs 0.2 +/- 0.9 in the Hyalgan and control groups respectively, P < 0.05). Deterioration in the structural parameters was less in the Hyalgan group, with a statistically significant difference for two of the three evaluated parameters (overall assessment of chondropathy: +5.1 +/- 12.7 vs 16.7 +/- 18.3, P = 0.016; SFA scoring system: +3.7 +/- 7.3 vs +9.0 +/- 11.5, P = 0.05) in the Hyalgan and control groups, respectively. CONCLUSIONS: This study supports existing data concerning the favorable symptomatic effect of intra-articular injections of Hyalgan in osteoarthritis of the knee and suggests that repeated intra-articular injections of Hyalgan might delay the structural progression of the disease. Other studies are required to confirm these results and to determine the long-term monitoring of osteoarthritic patients using such local therapy. PMID- 9219679 TI - The effect of strenuous versus moderate exercise on the metabolism of proteoglycans in articular cartilage from different weight-bearing regions of the equine third carpal bone. AB - Articular cartilage degeneration in the middle carpal joint is a common problem in racing horses. This study evaluated the effect of exercise on the in-vitro synthesis of the large aggregating proteoglycans (aggrecan) and two small proteoglycans, biglycan and decorin, in articular cartilage taken from three weight bearing regions of the third carpal bone of horses which were subjected to moderate or strenuous exercise. Twelve Standardbred horses free from clinical and radiographic disease of the middle carpal joint were subjected to an 8 week moderate exercise program. The horses were then randomly assigned to two groups: group A--continued moderate exercise and group B--strenuous exercise for 17 weeks. Horses were then rested for 16 weeks. Full-depth articular cartilage explants from the dorsal radial facet (DRF), dorsal intermediate facet (DIF) and palmar condyle (PC) of the third carpal bone were collected and cultured. Cartilage proteoglycan content and release into culture media were measured. Newly synthesized proteoglycans were labeled with 35SO4(2-) for 48 h and analyzed by size exclusion and hydrophobic chromatography, sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) and autoradiography. Histologic sections of adjacent osteochondral regions were evaluated for evidence of arthritic change. No histologic abnormalities or differences in proteoglycan content were detected in any of the articular cartilage regions examined. There was however, a significant reduction (P < 0.05) in aggrecan synthesis and a concomitant increase in decorin synthesis (P < 0.05) in articular cartilage from the DRF of group B animals. There was no change in biglycan synthesis, aggrecan hydrodynamic size or ability to aggregate in any articular cartilage region. This study has demonstrated that strenuous exercise in horses can lead to a disturbance in the biosynthesis of proteoglycans in articular cartilage regions subjected to high contact stresses (DRF). These metabolic abnormalities, which persisted for 16 weeks after cessation of exercise, could have deleterious effects on the biomechanical properties of the tissue. We suggest that the observed alteration in articular cartilage metabolism in CRF cartilage of strenuously exercised horses could represent a predisposing factor for cartilage degeneration and osteoarthritis at a later stage. PMID- 9219681 TI - Activation of fibrillar collagen synthesis and phenotypic modulation of chondrocytes in early human osteoarthritic cartilage lesions. AB - The objective of this study was to investigate the expression and extracellular distribution of fibrillar collagen types I, II, and III in early stage osteoarthritic cartilage in order to elucidate matrix gene expression and cell differentiation in early phases of the disease. Arthroscopically, derived specimens of early stage osteoarthritic articular cartilage were analyzed by histochemistry, immunohistochemistry and by in situ hybridization and compared with normal articular cartilage samples. In normal articular cartilage no significant mRNA expression of any of the investigated collagen types was found. In early stage osteoarthritic specimens, a strongly enhanced mRNA expression of the major cartilage matrix component type II collagen was detected. Additionally, a focal onset of type III, but not type I collagen expression was observed. Thus, besides activation of matrix synthesis, the modulation of the chondrocytic phenotype is likely to play a distinct role in the cellular response in the early phases of the degenerative process in osteoarthritis. PMID- 9219680 TI - Serial kinematic analysis of the canine hindlimb joints after deafferentation and anterior cruciate ligament transection. AB - OBJECTIVE AND DESIGN: Transection of the anterior cruciate ligament 2 weeks after ipsilateral hindlimb deafferentation leads to osteoarthritis of the knee joint within 3 weeks. We analyzed the gait of six dogs that underwent this procedure in order to identify kinematic changes that could account for this rapid joint degeneration. All animals were video taped, 1, 3, 6, 9 and 13 weeks after surgery while they trotted on a treadmill. RESULTS: In each dog, extension of the hip, knee and ankle joints of the unstable limb was increased, and the yield phase of the unstable knee was delayed or attenuated. When killed, five of six dogs showed a large full-thickness cartilage ulcer on the distal and/or anterior surface of the medial femoral condyle of the unstable knee; in the sixth dog, a smaller ulcer was observed. However, the severity of pathology in each individual was not obviously related to difference among the dogs in postoperative joint kinematics. CONCLUSIONS: These data, and results of prior studies in humans and dogs, suggest that knee hyperextension resulting from limb deafferentation, and knee instability resulting from anterior cruciate ligament transection, operate in concert to create a mechanical environment (i.e., increased tibiofemoral separation and changes in the loading of articular surfaces) that results in rapid joint breakdown. PMID- 9219682 TI - Immunolocalization of type IX collagen in normal and spontaneously osteoarthritic canine tibial cartilage and isolated chondrons. AB - OBJECTIVE: The pericellular localization of type IX collagen in avian and mammalian hyaline cartilages remains controversial, while its distribution during osteoarthritic degeneration is poorly understood. This study aimed to compare and contrast the immunohistochemical distribution of type IX collagen in normal mature and spontaneously osteoarthritic canine tibial cartilage. DESIGN: Thick vibratome sectioning techniques were evaluated and compared with isolated chondrons using a range of streptavidin-linked probes in combination with light, confocal and transmission electron microscopy. RESULTS: In normal intact samples, type IX collagen was concentrated in the pericellular microenvironment, while a weaker extracellular reaction around each chondron separated the territorial matrix from the unstained interterritorial matrix. Further differentiation was evident in isolated chondrons where the fibrous pericellular capsule stained more intensely than the tail and interconnecting segments between columnated chondrons. Two regions of type IX reactivity were identified in osteoarthritic tissue: an intensely stained superficial reactive region below the eroding margins, and normal deep layer cartilage where pericellular staining persists. The superficial reactive region was characterized by chondron swelling and chondrocyte cluster formation, a loss of pericellular type IX staining, and a significant increase in matrix staining between clusters. Disintegration and loss of fibrillar collagens was evident in both the swollen microenvironment and adjacent territorial matrices. CONCLUSIONS: The results suggest that changes in type IX distribution, expansion of the pericellular microenvironment and chondrocyte proliferation represent key elements in the chondron remodeling and chondrocyte cluster formation associated with osteoarthritic degeneration. PMID- 9219683 TI - Analysis of types I, II, III, IX and XI collagens synthesized by fetal bovine chondrocytes in high-density culture. AB - OBJECTIVE: This study was undertaken in order to determine phenotypic modulation of the chondrocytes more closely in high-density culture conditions and to clarify the role of ascorbate. Levels of five collagen types were analyzed qualitatively and quantitatively, and their distribution was observed in the cell layer and the culture medium. DESIGN: Types I, II, III, IX and XI collagens, synthesized by fetal bovine chondrocytes in high-density culture, were analyzed qualitatively and quantitatively by direct measurement of radiolabeled collagens separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by specific radioimmunoassays. RESULTS: Under the experimental conditions used in this study (0.6 x 10(6) cells/cm2), chondrocytes did not proliferate in the absence of ascorbate, whereas a twofold increase in cell number was observed in the presence of ascorbate at day 14. Cartilage-specific collagens (types II, IX and XI) were synthesized throughout the culture period (up to 47 days), as was type III collagen, which appeared as early as day 1 and was essentially present in the culture medium. Partial dedifferentiation of chondrocytes was demonstrated by the synthesis of type I collagen, which was detected by day 2 in culture medium containing ascorbate, and by day 6 without ascorbate. After 33 days of culture, a threefold increase in type I collagen synthesis was observed in culture medium with ascorbate, reaching 66% of the type II collagen content of the cell layer. One month of culture marked the onset of a progressive decrease in the synthesis of all collagen types. CONCLUSIONS: Under these high-density culture conditions, fetal bovine chondrocytes undergo a time and ascorbate dependent program of partial dedifferentiation. This system provides a simple model for studying the initial mechanisms of chondrocytes dedifferentiation. PMID- 9219684 TI - The Ras-RasGAP complex: structural basis for GTPase activation and its loss in oncogenic Ras mutants. AB - The three-dimensional structure of the complex between human H-Ras bound to guanosine diphosphate and the guanosine triphosphatase (GTPase)-activating domain of the human GTPase-activating protein p120GAP (GAP-334) in the presence of aluminum fluoride was solved at a resolution of 2.5 angstroms. The structure shows the partly hydrophilic and partly hydrophobic nature of the communication between the two molecules, which explains the sensitivity of the interaction toward both salts and lipids. An arginine side chain (arginine-789) of GAP-334 is supplied into the active site of Ras to neutralize developing charges in the transition state. The switch II region of Ras is stabilized by GAP-334, thus allowing glutamine-61 of Ras, mutation of which activates the oncogenic potential, to participate in catalysis. The structural arrangement in the active site is consistent with a mostly associative mechanism of phosphoryl transfer and provides an explanation for the activation of Ras by glycine-12 and glutamine-61 mutations. Glycine-12 in the transition state mimic is within van der Waals distance of both arginine-789 of GAP-334 and glutamine-61 of Ras, and even its mutation to alanine would disturb the arrangements of residues in the transition state. PMID- 9219685 TI - Crystal structure of mouse CD1: An MHC-like fold with a large hydrophobic binding groove. AB - CD1 represents a third lineage of antigen-presenting molecules that are distantly related to major histocompatibility complex (MHC) molecules in the immune system. The crystal structure of mouse CD1d1, corresponding to human CD1d, at 2.8 resolution shows that CD1 adopts an MHC fold that is more closely related to that of MHC class I than to that of MHC class II. The binding groove, although significantly narrower, is substantially larger because of increased depth and it has only two major pockets that are almost completely hydrophobic. The extreme hydrophobicity and shape of the binding site are consistent with observations that human CD1b and CD1c can present mycobacterial cell wall antigens, such as mycolic acid and lipoarabinomannans. However, mouse CD1d1 can present very hydrophobic peptides, but must do so in a very different way from MHC class Ia and class II molecules. PMID- 9219686 TI - Water on the sun: line assignments based on variational calculations. AB - The infrared spectrum of hot water observed in a sunspot has been assigned. The high temperature of the sunspot (3200 K) gave rise to a highly congested pure rotational spectrum in the 10-micrometer region that involved energy levels at least halfway to dissociation. Traditional spectroscopy, based on perturbation theory, is inadequate for this problem. Instead, accurate variational solutions of the vibration-rotation Schrodinger equation were used to make assignments, revealing unexpected features, including rotational difference bands and fewer degeneracies than anticipated. These results indicate that a shift away from perturbation theory to first principles calculations is necessary in order to assign spectra of hot polyatomic molecules such as water. PMID- 9219689 TI - The ionosphere of Europa from Galileo radio occultations. AB - The Galileo spacecraft performed six radio occultation observations of Jupiter's Galilean satellite Europa during its tour of the jovian system. In five of the six instances, these occultations revealed the presence of a tenuous ionosphere on Europa, with an average maximum electron density of nearly 10(4) per cubic centimeter near the surface and a plasma scale height of about 240 +/- 40 kilometers from the surface to 300 kilometers and of 440 +/- 60 kilometers above 300 kilometers. Such an ionosphere could be produced by solar photoionization and jovian magnetospheric particle impact in an atmosphere having a surface density of about 10(8) electrons per cubic centimeter. If this atmosphere is composed primarily of O2, then the principal ion is O2+ and the neutral atmosphere temperature implied by the 240-kilometer scale height is about 600 kelvin. If it is composed of H2O, the principal ion is H3O+ and the neutral temperature is about 340 kelvin. In either case, these temperatures are much higher than those observed on Europa's surface, and an external heating source from the jovian magnetosphere is required. PMID- 9219690 TI - Experimental simulations of sulfide formation in the solar nebula. AB - Sulfurization of meteoritic metal in H2S-H2 gas produced three different sulfides: monosulfide solid solution [(Fe,Ni)1-xS], pentlandite [(Fe,Ni)9-xS8], and a phosphorus-rich sulfide. The composition of the remnant metal was unchanged. These results are contrary to theoretical predictions that sulfide formation in the solar nebula produced troilite (FeS) and enriched the remaining metal in nickel. The experimental sulfides are chemically and morphologically similar to sulfide grains in the matrix of the Alais (class CI) carbonaceous chondrite, suggesting that these meteoritic sulfides may be condensates from the solar nebula. PMID- 9219693 TI - Precursor-directed biosynthesis of erythromycin analogs by an engineered polyketide synthase. AB - A genetic block was introduced in the first condensation step of the polyketide biosynthetic pathway that leads to the formation of 6-deoxyerythronolide B (6 dEB), the macrocyclic precursor of erythromycin. Exogenous addition of designed synthetic molecules to small-scale cultures of this null mutant resulted in highly selective multimilligram production of unnatural polyketides, including aromatic and ring-expanded variants of 6-dEB. Unexpected incorporation patterns were observed, illustrating the catalytic versatility of modular polyketide synthases. Further processing of some of these scaffolds by postpolyketide enzymes of the erythromycin pathway resulted in the generation of novel antibacterials with in vitro potency comparable to that of their natural counterparts. PMID- 9219694 TI - Inhibition of Bax channel-forming activity by Bcl-2. AB - Proteins of the Bcl-2 family are intracellular membrane-associated proteins that regulate programmed cell death (apoptosis) either positively or negatively by as yet unknown mechanisms. Bax, a pro-apoptotic member of the Bcl-2 family, was shown to form channels in lipid membranes. Bax triggered the release of liposome encapsulated carboxyfluorescein at both neutral and acidic pH. At physiological pH, release could be blocked by Bcl-2. Bcl-2, in contrast, triggered carboxyfluorescein release at acidic pH only. In planar lipid bilayers, Bax formed pH- and voltage-dependent ion-conducting channels. Thus, the pro-apoptotic effects of Bax may be elicited through an intrinsic pore-forming activity that can be antagonized by Bcl-2. PMID- 9219695 TI - Alternative cleavage of Alzheimer-associated presenilins during apoptosis by a caspase-3 family protease. AB - Most cases of early-onset familial Alzheimer's disease (FAD) are caused by mutations in the genes encoding the presenilin 1 (PS1) and PS2 proteins, both of which undergo regulated endoproteolytic processing. During apoptosis, PS1 and PS2 were shown to be cleaved at sites distal to their normal cleavage sites by a caspase-3 family protease. In cells expressing PS2 containing the asparagine-141 FAD mutant, the ratio of alternative to normal PS2 cleavage fragments was increased relative to wild-type PS2-expressing cells, suggesting a potential role for apoptosis-associated cleavage of presenilins in the pathogenesis of Alzheimer's disease. PMID- 9219696 TI - Differential effects of early hippocampal pathology on episodic and semantic memory. AB - Global anterograde amnesia is described in three patients with brain injuries that occurred in one case at birth, in another by age 4, and in the third at age 9. Magnetic resonance techniques revealed bilateral hippocampal pathology in all three cases. Remarkably, despite their pronounced amnesia for the episodes of everyday life, all three patients attended mainstream schools and attained levels of speech and language competence, literacy, and factual knowledge that are within the low average to average range. The findings provide support for the view that the episodic and semantic components of cognitive memory are partly dissociable, with only the episodic component being fully dependent on the hippocampus. PMID- 9219697 TI - In vitro propagation of the prion-like state of yeast Sup35 protein. AB - The yeast cytoplasmically inherited genetic determinant [PSI+] is presumed to be a manifestation of the prion-like properties of the Sup35 protein (Sup35p). Here, cell-free conversion of Sup35p from [psi-] cells (Sup35ppsi-) to the prion-like [PSI+]-specific form (Sup35pPSI+) was observed. The conversion reaction could be repeated for several consecutive cycles, thus modeling in vitro continuous [PSI+] propagation. Size fractionation of lysates of [PSI+] cells demonstrated that the converting activity was associated solely with Sup35pPSI+ aggregates, which agrees with the nucleation model for [PSI+] propagation. Sup35pPSI+ was purified and showed high conversion activity, thus confirming the prion hypothesis for Sup35p. PMID- 9219698 TI - Mating type switching in yeast controlled by asymmetric localization of ASH1 mRNA. AB - Cell divisions that produce progeny differing in their patterns of gene expression are key to the development of multicellular organisms. In the budding yeast Saccharomyces cerevisiae, mother cells but not daughter cells can switch mating type because they selectively express the HO endonuclease gene. This asymmetry is due to the preferential accumulation of an unstable transcriptional repressor protein, Ash1p, in daughter cell nuclei. Here it is shown that ASH1 messenger RNA (mRNA) preferentially accumulates in daughter cells by a process that is dependent on actin and myosin. A cis-acting element in the 3' untranslated region of ASH1 mRNA is sufficient to localize a chimeric RNA to daughter cells. These results suggest that localization of mRNA may have been an early property of the eukaryotic lineage. PMID- 9219699 TI - Treatment of rheumatoid arthritis with a recombinant human tumor necrosis factor receptor (p75)-Fc fusion protein. AB - BACKGROUND: Tumor necrosis factor (TNF) is a proinflammatory cytokine involved in the pathogenesis of rheumatoid arthritis, and antagonism of TNF may reduce the activity of the disease. This study evaluated the safety and efficacy of a novel TNF antagonist - a recombinant fusion protein that consists of the soluble TNF receptor (p75) linked to the Fc portion of human IgG1 (TNFR:Fc). METHODS: In this multicenter, double-blind trial, we randomly assigned 180 patients with refractory rheumatoid arthritis to receive subcutaneous injections of placebo or one of three doses of TNFR:Fc (0.25, 2, or 16 mg per square meter of body-surface area) twice weekly for three months. The clinical response was measured by changes in composite symptoms of arthritis defined according to American College of Rheumatology criteria. RESULTS: Treatment with TNFR:Fc led to significant reductions in disease activity, and the therapeutic effects of TNFR:Fc were dose related. At three months, 75 percent of the patients in the group assigned to 16 mg of TNFR:Fc per square meter had improvement of 20 percent or more in symptoms, as compared with 14 percent in the placebo group (P<0.001). In the group assigned to 16 mg per square meter, the mean percent reduction in the number of tender or swollen joints at three months was 61 percent, as compared with 25 percent in the placebo group (P<0.001). The most common adverse events were mild injection-site reactions and mild upper respiratory tract symptoms. There were no dose-limiting toxic effects, and no antibodies to TNFR:Fc were detected in serum samples. CONCLUSIONS: In this three-month trial TNFR:Fc was safe, well tolerated, and associated with improvement in the inflammatory symptoms of rheumatoid arthritis. PMID- 9219700 TI - Prednisone and aspirin in women with autoantibodies and unexplained recurrent fetal loss. AB - BACKGROUND: Recurrent fetal loss has been well described in women with antiphospholipid antibodies. Such women also often have other autoantibodies commonly found in patients with systemic lupus erythematosus. Treating them with prednisone and aspirin may reduce the risk of fetal loss. METHODS: We screened 773 nonpregnant women who had the unexplained loss of at least two fetuses for antinuclear, anti-DNA, antilymphocyte, and anticardiolipin antibodies and for the lupus anticoagulant. Of 385 women with at least one autoantibody, 202 who later became pregnant were randomly assigned in equal numbers to receive either prednisone (0.5 to 0.8 mg per kilogram of body weight per day) and aspirin (100 mg per day) or placebo for the duration of the pregnancy. The women were stratified according to age (18 to 34 years or 35 to 39 years) and the week of gestation at which the previous fetal losses had occurred (< or = 12 or > 12 weeks). The primary outcome measure was a successful pregnancy. RESULTS: Live infants were born to 66 women in the treatment group (65 percent) and 57 women in the placebo group (56 percent, P=0.19). More infants were born prematurely in the treatment group than in the placebo group (62 percent vs. 12 percent, P<0.001). The major side effects of therapy in the mothers were hypertension (treatment group, 13 percent; placebo group, 5 percent; P=0.05) and diabetes mellitus (15 percent and 5 percent, P=0.02). CONCLUSIONS: Treating women who have autoantibodies and recurrent fetal loss with prednisone and aspirin is not effective in promoting live birth, and it increases the risk of prematurity. PMID- 9219701 TI - Pregnancy loss in the antiphospholipid-antibody syndrome--a possible thrombogenic mechanism. AB - BACKGROUND: The mechanisms of vascular thrombosis and pregnancy loss in the antiphospholipid-antibody syndrome are unknown. Levels of annexin V, a phospholipid-binding protein with potent anticoagulant activity, are markedly reduced on placental villi from women with this syndrome. Hypercoagulability in such women may therefore be due to the reduction of surface-bound annexin V by antiphospholipid antibodies. To test this idea, we studied how antiphospholipid antibodies affect levels of annexin V on cultured trophoblasts and human umbilical-vein endothelial cells and how they affect the procoagulant activity of these cells. METHODS: We isolated IgG fractions from three patients with the antiphospholipid-antibody syndrome and from normal controls. These antibodies were incubated with cultured BeWo cells (a placental-trophoblast cell line), primary cultured trophoblasts, and human umbilical-vein endothelial cells. Annexin V on the cell surfaces was measured by an enzyme-linked immunosorbent assay. The coagulation times of plasma overlaid on the cells were also determined. RESULTS: Trophoblasts and endothelial cells exposed to antiphospholipid-antibody IgG as compared with control IgG had reduced levels of annexin V (trophoblasts, 0.37 +/- 0.02 vs. 0.85 +/- 0.12 ng per well, P=0.02; endothelial cells, 1.6 +/- 0.04 vs. 2.1 +/- 0.05 ng per well, P=0.001). Also, trophoblasts and endothelial cells exposed to antiphospholipid-antibody IgG had faster mean (+/- SE) plasma coagulation times than cells exposed to control IgG (trophoblasts, 8.7 +/- 2.0 vs. 21.3 +/- 2.9 minutes, P=0.02; endothelial cells, 9.8 +/- 0.8 vs. 14.2 +/- 1.2 minutes, P=0.04). CONCLUSIONS: Antiphospholipid antibodies reduce the levels of annexin V and accelerate the coagulation of plasma on cultured trophoblasts and endothelial cells. The reduction of annexin V levels on vascular cells may be an important mechanism of thrombosis and pregnancy loss in the antiphospholipid-antibody syndrome. PMID- 9219702 TI - Chemoradiotherapy followed by surgery compared with surgery alone in squamous cell cancer of the esophagus. AB - BACKGROUND: We conducted a multicenter, randomized trial to compare preoperative chemoradiotherapy followed by surgery with surgery alone in patients with stage I and II squamous-cell cancer of the esophagus. METHODS: The preoperative combined therapy consisted of two one-week courses; each involved radiotherapy, in a dose of 18.5 Gy delivered in five fractions of 3.7 Gy each, and 80 mg of cisplatin per square meter of body-surface area, administered 0 to 2 days before the first day of radiotherapy. The surgical plan included one-stage en bloc esophagectomy and proximal gastrectomy by the abdominal and right thoracic routes, to be performed immediately after randomization in the group assigned to surgery alone and two to four weeks after the completion of preoperative chemoradiotherapy in the group assigned to combined therapy. RESULTS: A total of 297 patients entered the study; 11 were found to be ineligible, and 4 were lost to follow-up. Of the remaining 282, 139 were assigned to surgery alone and 143 to combined therapy. After a median follow-up of 55.2 months, no significant difference in overall survival was observed; the median survival was 18.6 months for both groups. As compared with the group treated with surgery alone, the group treated preoperatively had longer disease-free survival (P=0.003), a longer interval free of local disease (P=0.01), a lower rate of cancer-related deaths (P=0.002), and a higher frequency of curative resection (P=0.017). However, there were more postoperative deaths (P=0.012) in the group treated preoperatively with chemoradiotherapy. Three prognostic factors were found to influence survival in a multivariate analysis: the disease stage, based on computed tomography; the location of the tumor; and whether the surgical resection was curative. CONCLUSIONS: In patients with squamous-cell esophageal cancer, preoperative chemoradiotherapy did not improve overall survival, but it did prolong disease-free survival and survival free of local disease. PMID- 9219703 TI - Images in clinical medicine. Nail changes after chemotherapy. PMID- 9219704 TI - The Medicare-HMO revolving door--the healthy go in and the sick go out. AB - BACKGROUND: Enrollment in Medicare health maintenance organizations (HMOs) is encouraged because of the expectation that HMOs can help slow the growth of Medicare costs. However, Medicare HMOs, which are paid 95 percent of average yearly fee-for-service Medicare expenditures, are increasingly believed to benefit from the selective enrollment of healthier Medicare recipients. Furthermore, whether sicker patients are more likely to disenroll from Medicare HMOs, thus raising average fee-for-service costs, is not clear. METHODS: We used Medicare enrollment and inpatient billing records for southern Florida from 1990 through 1993 to examine differences in the use of inpatient medical services by 375,406 beneficiaries in the Medicare fee-for-service system, 48,380 HMO enrollees before enrollment, and 23,870 HMO enrollees after disenrollment. We also determined whether these differences were related to demographic characteristics and whether the pattern of use after disenrollment persisted over time. RESULTS: The rate of use of inpatient services in the HMO-enrollment group during the year before enrollment was 66 percent of the rate in the fee-for service group, whereas the rate in the HMO-disenrollment group after disenrollment was 180 percent of that in the fee-for-service group. Beneficiaries who disenrolled from HMOs re-enrolled at about the time that their level of use dropped to that in the fee-for-service group. CONCLUSIONS: These data show marked selection biases with respect to HMO enrollment and disenrollment. These biases undermine the effectiveness of the Medicare managed-care system and highlight the need for longitudinal and population-based studies. PMID- 9219705 TI - Insulin lispro. PMID- 9219706 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 22-1997. A 58-year-old woman with multiple cranial neuropathies. PMID- 9219708 TI - Anticytokine therapy in rheumatoid arthritis. PMID- 9219707 TI - Fixing Medicare. PMID- 9219709 TI - Autoantibodies and pregnancy loss. PMID- 9219710 TI - Patients' rights in managed care--exit, voice, and choice. PMID- 9219711 TI - MR imaging of the elbow. Technical considerations. AB - The elbow has proven the most technically challenging of all of the major joints in MR imaging, particularly with regard to radiofrequency design. It is possible to obtain high-quality diagnostic studies on most clinical MR imaging systems that are made today using available coils and pulse sequences. This article evaluates techniques for MR imaging of the elbow. PMID- 9219712 TI - The role of MR imaging in the management of elbow problems. AB - In the past several years, the role of MR imaging in diagnosing pathologic conditions of the elbow has dramatically increased. Aside from imaging soft tissue tumors, it can accurately visualize partial and complete tears of tendons and ligaments, as well as displacement of epiphyseal fractures in children. Its role in identifying loose bodies, particularly when they are nonosseous, and areas of osteochondritis dissecans has also increased. The use of MR imaging for diagnosing neuropathies, particularly when electrodiagnostic studies are negative, offers exciting possibilities as additional technical improvements are developed. PMID- 9219713 TI - Normal MR imaging anatomy of the elbow. AB - This article discusses the normal, clinically relevant MR imaging anatomy of the elbow. A compartmental approach is utilized to help simplify this anatomically complex region. Imaging techniques, common anatomic variants, and imaging pitfalls are also briefly discussed. PMID- 9219714 TI - MR imaging of normal variants and interpretation pitfalls of the elbow. AB - Discrimination of normal anatomic landmarks from true disease is one of the fundamental tenets of adept MR imaging. The radiologist is thus compelled to accumulate a comprehensive knowledge of normal structures, variants, and potential MR imaging interpretation pitfalls. In this article the authors focus on a number of normal, bony, ligamentous, and tendinous structures that can simulate disease at the elbow. A discussion of the particular anatomy responsible for the appearance of each of these interpretation pitfalls is provided. In addition, ways to distinguish these pitfalls from true elbow disease are discussed. PMID- 9219715 TI - MR imaging of the normal developmental anatomy of the elbow. AB - Growth and ossification of the elbow are complex. Interpreting MR images of the elbow in children requires a knowledge of the elbow's developmental changes. This article discusses the basic principles of growth and development of the distal humerus, proximal radius, and ulna. The signal characteristics of cartilage and marrow in the immature skeleton are described. Technical factors related to imaging of growth cartilage are outlined, and specific challenges during imaging of the pediatric elbow are emphasized. The changing MR appearance of the elbow due to ossification and physeal closure is described. The article also explains several pitfalls encountered. PMID- 9219716 TI - MR imaging of ligamentous abnormalities of the elbow. AB - With high-resolution MR imaging, the collateral ligament of the elbow can be directly evaluated; thus ligamentous injuries can be detected, localized, and graded. The functional anatomy of the ligaments of the elbow is reviewed. The normal appearance of the ligaments on MR images as well as the clinical and MR imaging findings of medial and lateral collateral ligament injury are presented. PMID- 9219717 TI - MR imaging of tendon injuries in the elbow. AB - Tendon injuries are among the more common sources of elbow pain, resulting from occupational and sports activities as well as the activities of daily living. The exquisite soft-tissue contrast resolution and multiplanar capability of MR imaging have proven ideal for evaluating, diagnosing, and managing tendon injuries. Familiarity with normal anatomy, the spectrum of tendon derangement from tendinosis to complete tears, and the typical injury mechanisms involved in tendinopathy about the elbow are all invaluable for appropriate MR imaging examination and interpretation. PMID- 9219718 TI - MR features of nerve disorders at the elbow. AB - MR imaging is a useful method for evaluating nerve disease. It can portray the normal anatomy and identify unsuspected space-occupying masses. Severe intrinsic nerve disease can be depicted. This article outlines the normal anatomy of the three major nerves that traverse the elbow joint: the ulnar nerve, the median nerve, and the radial nerve. Entrapment and compression neuropathies of each nerve are discussed in detail. Finally, the role of MR imaging in delineating each nerve abnormality is examined. PMID- 9219719 TI - MR imaging of pediatric elbow fractures. AB - Pediatric elbow fractures are elusive to radiographic detection and more sophisticated imaging techniques are often required to assess the presence and extension of the fracture line to make appropriate treatment decisions. Arthrography is the current method of choice, but ultrasonography and, more recently, MR imaging have been proposed as alternative modalities, although further experience is necessary to validate their usefulness. We believe that MR imaging offers significant benefits because of its exquisite spatial and contrast resolution. PMID- 9219720 TI - MR imaging of osteochondral and articular lesions. AB - MR imaging provides clinically useful information in assessing the elbow joint for osteochondral and articular lesions. Post-traumatic osseous abnormalities well seen by MR imaging include radiographically occult fractures, stress fractures, bone contusions, osteochondritis dissecans, and chondral defects. Intraarticular loose bodies can be identified with MR imaging, especially if fluid or contrast material are present within the elbow joint. MR imaging can also provide additional information regarding synovial osteochondritis, osteoarthritis, and bursitis about the elbow. PMID- 9219721 TI - Arthropathies and inflammatory conditions of the elbow. AB - MR imaging of arthropathies and inflammatory conditions affecting the elbow are presented. Noninfectious conditions discussed include osteoarthritis, disorders characterized by synovial proliferation, pigmented villo- nodular synovitis, synovial osteochondromatosis, crystal deposition disorders, and neuropathic osteoarthropathy. Infectious conditions discussed include septic olecranon bursitis, septic arthritis, osteomyelitis, and pyomyositis. Clinical aspects of these entities are discussed, including utility of MR imaging for diagnosis and clinical management. PMID- 9219722 TI - MR imaging of musculoskeletal tumors in the elbow region. AB - MR imaging is useful in the evaluation of benign and malignant osseous and soft tissue masses in the elbow. It allows for multiplanar, high-contrast images that provide precise definition of local tumor extent. The paucity of fat and a high concentration of small soft-tissue structures such as tendons, ligaments, and nerves in this area makes MR imaging more advantageous than CT for delineating tumor margins in the soft tissue. This article describes some of the more common tumors around the elbow with emphasis on their MR imaging characteristics. PMID- 9219723 TI - Western immunoblotting and enzymatic activity analysis of cathepsin D in human breast cancer cell lines of different invasive potential. Regulation by 17beta estradiol, tamoxifen and ICI 182,780. AB - We used enzymatic activity and immunochemical quantifications to analyse the expression and secretion of cathepsin D by human breast cancer cell lines of different invasive potentials (MCF-7/6, MCF-7/AZ, MDA-MB-231). This study does not directly prove that cathepsin D or procathepsin D is involved in human breast cancer cell invasion and metastasis but it shows that the proportion of procathepsin D (activity and antigen) secreted by the human breast cancer cell lines tested correlates with their invasive potential. In the estrogen receptor positive MCF-7 subclones, this proportion is increased by estradiol only in the invasive MCF-7/6 variant. The cell content in procathepsin D is increased by estrogens to a greater extent in MCF-7/6 cells as compared to non-invasive MCF 7/AZ cells. Tamoxifen appears to be an estrogen agonist concerning cathepsin D regulation, whereas ICI 182,780 is a true antagonist. Our results suggest that synthesis and secretion of cathepsin D are regulated at two distinct levels and differentially affected by estrogens. Synthesis only seems to be affected in non invasive MCF-7/AZ cells, whereas in invasive MCF-7/6 cells, both synthesis and the efficiency of secretion are increased by estrogens. Our results also confirm that the key site of regulation leading to lysosomal enzyme oversecretion is the Golgi apparatus insulin-like growth factor-II/mannose 6-phosphate receptor. PMID- 9219724 TI - Effect of cisplatin and BCNU on MMP-2 levels in human glioblastoma cell lines in vitro. AB - Matrix metalloproteinases (MMPs) play an important role in various physiological and pathological conditions such as tissue remodeling, and cancer cell invasion and metastasis. The aim of this study was to determine the effect of the antitumor compounds cis-dichlorodiammine platinum (ii) (cisplatin) and 1, 3 bis (2-chloroethyl)-1-nitrosourea (BCNU) on 72-kDa type IV collagenase activity (MMP 2) in human gliomas. Human glioblastoma cell lines were treated with cisplatin (25 microM), and BCNU (50 microM), and the levels of MMP-2 were estimated in serum-free conditioned medium and in cell extracts at different time intervals. Gelatin zymography revealed increased levels of MMP-2 in serum-free conditioned medium and in cell extracts of untreated glioblastoma cell cultures during a 72-h period. In contrast, MMP-2 levels were significantly decreased in cisplatin treated cells both in conditioned medium and cell extracts. However, no significant changes of MMP-2 levels were noted in BCNU-treated cells. Quantitative analysis of MMP-2 enzyme activity by densitometry and amount of MMP 2 protein by ELISA showed significantly decreased levels of MMP-2 in cisplatin treated cells compared to BCNU and untreated glioblastoma cells. The results indicate that decreased levels of MMP-2 might represent an additional mechanism by which cisplatin provides its antineoplastic effects. PMID- 9219725 TI - Cathepsin B and cysteine proteinase inhibitors in human lung cancer cell lines. AB - Cell lines derived from human squamous cell (EPCL), large cell (LCLC), and small cell lung cancer (SCLC) lines were investigated for the expression of cathepsin B (Cat B) and cysteine proteinase inhibitors (CPIs). The EPLC and LCLC lines expressed 5- to 50-fold more Cat B activity and contained more mature Cat B of M(r) 27-29 kDa (> 2.5 microg/mg total protein) than the SCLC lines (< 1.0 microg/mg total protein). The LPLC lines also secreted the highest amounts of Cat B precursor of M(r) about 46 kDa. Inhibitory activities against Cat B and papain were associated with high molecular mass (HMM) and low molecular mass (LMM) inhibitory proteins, both in cell extracts and in media. About 75% of the inhibitory activity was associated with HMM inhibitors, the majority of which were kininogens (M(r) > or = 67 kDa). The LMM inhibitors of M(r) 10-15 kDa were cystatin C and stefins A and B, which were quantitated by ELISA: stefins A and B were present in cell extracts and medium in similar concentrations (5-200 ng/10(6) cells), while 80-99% of the cystatin C was released in the medium (10 195 ng/10(6) cells). Phorbol ester (PMA), which induces protein-kinase C mediated signal transduction and enhances cellular differentiation in many non-small cell lung cancer (NSCLC) cell lines, increased intracellular Cat B activity and Cat B protein as well as its secretion in some cell lines but not in others, regardless of their histological type. PMA significantly (P < 0.049) decreased intracellular stefin A concentrations in two EPLC lines and non-significantly in two LCLC lines. PMA decreased secretion of stefin A in all EPLC lines, but not in LCLC lines, while IGF-I significantly increased stefin B secretion in both SCLC lines. These data showed that lung tumor cells produce both cysteine proteinases and cystatins. As the antagonistic molecules are regulated differently in histologically different types of lung tumor cells, it is possible that an imbalance between the proteinases and their specific inhibitors plays a role in progression of certain types of lung tumors in vivo. PMID- 9219726 TI - Breast cancer cells have a high capacity to acidify extracellular milieu by a dual mechanism. AB - The extracellular pH in malignant tumors is known to be lower than in normal tissues and may therefore facilitate extracellular activation of secreted lysosomal cathepsins. We have tested the capability of human mammary cells (continuous cell lines and primary culture) to acidify their extracellular environment, using two techniques. By measuring pH changes through alterations of phenolsulfone phthaleine absorbance, we found that the more aggressive MDA-MB-231 human breast cancer cells were more active in acidifying a non-buffered balanced salt solution than the estrogen receptor positive MCF7 and ZR75 cell lines and than normal mammary epithelial cells in primary culture. Metastatic breast cancer cells from pleural effusions were up to 200-fold more active in acidifying their extracellular milieu than non-malignant mammary cells cultured in the same conditions, strongly suggesting that this difference also occurs in vivo. The use of inhibitors in the presence or absence of glucose showed that both lactate and an ATP-driven proton pump sharing some characteristics of the vacuolar H+ pump were involved. Bafilomycin A1, a specific inhibitor of the vacuolar (V-type) ATP H+ pump inhibited part of the acidification by MCF7 cells, but not by MDA-MB-231 cells. We also used microelectrodes to measure extracellular pH, in close contact to the MCF7 breast cancer cells. The pH at the free surface of MCF7 cells was lower by 0.33 +/- 0.14 unit than that of the surrounding medium, while insertion of the microelectrode tip beneath the attached surface of the cells showed a greater lowering of pH from 0.3 to 1.7 pH unit as long as cell attachment on the substrate prevented H+ diffusion. We conclude that breast carcinoma cells have a higher capacity for acidifying their extracellular milieu than normal mammary cells, and that both a plasma membrane H(+)-ATPase, and lactic acid production are involved in this acidification. It is therefore possible that the aspartyl and cysteinyl pro-cathepsins secreted in excess by tumor cells may be activated extracellularly in vivo close to the basement membrane. PMID- 9219727 TI - Influence of antiestrogens on the migration of breast cancer cells using an in vitro wound model. AB - The metastasis of malignant tumor cells to other organs in the body is the major cause of cancer-related patient mortality. Therefore, the inhibition of tumor cell motility is critical in the prevention or control of tumor malignancy. In the present study, the antimetastatic potential of antiestrogens [tamoxifen (TAM); ICI-182,780 (ICI); and Analog II (AII)] on highly invasive, estrogen receptor (ER)-negative MDA-MB-231 (MDA) and non-invasive, ER-positive MCF-7 (MCF) human breast cancer cell lines was investigated using an in vitro wound model. Wounds were created in confluent cell cultures and repopulation of the wound space was evaluated by counting the number of cells that migrated into the wound area and by measuring the maximum distance traveled. In addition, the number of cells that were passively seeded into the wounded area was determined. ICI and AII reduced the number of MCF cells that migrated into the wounded area and reduced the number of viable passively seeded MDA cells. Unlike ICI and AII, TAM appeared to enhance MCF and MDA cell movement. This study indicates that the in vitro wound technique is applicable to the study of breast cancer cell movement in response to antiestrogens and other antimetastatic agents. It also demonstrates that antiestrogens differ in their influence on breast cancer cell migration. PMID- 9219728 TI - Regulation of MMP-9 (92 kDa type IV collagenase/gelatinase B) expression in stromal cells of human giant cell tumor of bone. AB - Matrix metalloproteinases (MMPs) play an important regulatory role in tissue morphogenesis, cell differentiation, tumor invasion and metastasis. Several authors have reported a direct correlation between the production of 72 kDa (MMP 2) and 92 kDa (MMP-9) type IV collagenases/gelatinases and the metastatic potential of cancer cells. Recently, we have identified the expression of both MMP-2 and MMP-9 in primary cultures of human giant cell tumor (GCT) of bone in vitro, and in tissue extracts in vivo. Interestingly, MMP-9 is not secreted by late-passaged GCT cells. It is possible that the production of MMP-9 is regulated by certain factor(s) secreted by the multinucleated giant cells in the primary culture. In order to test this hypothesis, the effect of primary-culture conditioned medium on the expression of MMP-9 by late-passaged mononuclear stromal cells was examined. Adding conditioned medium from the primary GCT culture to the late-passaged stromal cells induced MMP-9, as evidenced by the presence of lytic bands at M(r) 92,000 and 72,000 on a gelatin zymogram. These enzyme activities were inhibited by EDTA, a well-known inhibitor of the MMPs. We confirmed these results by Western blotting using specific antibodies and RT-PCR for MMP-2 and MMP-9. Immunofluorescence studies with specific antibodies to MMP-9 further confirmed its expression by the passaged stromal cells cultured in the primary-culture-conditioned medium. The data indicate that MMP-2 and MMP-9 are produced by the mononuclear stromal cells when cultured in GCT primary-culture conditioned medium. This suggests that multinucleated giant cells in primary cultures secrete a factor(s) that stimulates stromal cells to produce MMP-9, which, in turn, may contribute to the aggressive behavior of GCT. PMID- 9219729 TI - Influence of lipid diets on the number of metastases and ganglioside content of H59 variant tumors. AB - We investigated the influence of the fatty acid composition of the diet on the number of hepatic metastases and the ganglioside profile of the primary tumor and metastases. C57BL/6 female mice were fed different diets containing either no fats (TEK) or 8% of fish oil (POL), linseed oil (LIN), safflower oil (SAF) or beef tallow (BT) and were injected subcutaneously in the dorsum with H59 cells, a variant of the Lewis lung carcinoma (3LLc) that metastasizes preferentially to the liver. The omega3 polyunsaturated fatty acid (PUFA)-rich diets (LIN and POL) elicited more metastases than the omega6 PUFA-rich (SAF), fat-free (TEK), or saturated fats (BT) diets. However, dietary fat did not influence the ganglioside composition of either the primary tumors or the metastases, at least in the glucidic part. However, comparison of diets with low (TEK, SAF, and BT) and high (LIN and POL) number of metastases showed that the levels of G3 (which could be a second band of GM2) were greater in metastases of the latter group. This study showed that the H59 hepatic metastases contained more GM2 than the s.c. tumors, irrespective of diet or the number of metastases produced. The small differences in the ganglioside profiles observed in this study could have resulted from the limitations of the HPTLC method. A detailed analysis of the lipid chains, as well as glycolipids other than gangliosides, could give more information on changes resulting from different lipid diets. PMID- 9219730 TI - Urokinase-type plasminogen activator receptor in gastric cancer: tissue expression and prognostic role. AB - The urokinase-type plasminogen activator (UPA) and its inhibitor PAI-1 are thought to play an important part in gastric cancer (GC) invasion and metastasis. Little is known about the behavior and prognostic impact of the receptor for UPA (UPAR). The aims of the present study were: (1) to measure UPAR, UPA and PAI-1 levels in GC and in non-malignant tissue distant from the tumor (NORM); (2) to evaluate their relationship with histomorphological parameters; and (3) to determine their prognostic value. UPAR, UPA and PAI-1 levels were determined by ELISA in GC and NORM samples from 20 patients with GC undergoing surgery. The GC was also examined in terms of the presence (n = 10) or absence (n = 10) of metastasis, differentiation (five differentiated, 15 undifferentiated) and histotype. Survival was analysed using life table analysis. UPAR, UPA and PAI-1 were significantly higher in GC vs NORM, in the presence of metastasis (UPAR, UPA) and in undifferentiated GC (UPAR, PAI-1). UPAR significantly correlated with UPA and PAI-1. Low levels of UPAR (P = 0.04), UPA (P = 0.007) and PAI-1 (P = 0.02) were associated with a better survival. Our results demonstrate a sharp increase in UPAR in GC and suggest a prognostic role for it. The concomitant activation of UPAR, UPA and PAI-1 in GC confirm the important role of the plasminogen activator system in the process of invasion and metastasis. PMID- 9219731 TI - Melanoma cell adhesion to injured arterioles: mechanisms of stabilized tethering. AB - An isolated perfused vessel model was used to examine the mechanisms underlying the adhesive interactions between circulating tumor cells and subendothelial matrix in denuded arterioles. Arterioles ranging from 70 to 100 microm in diameter were isolated from rat mesentery, transferred to an isolated vessel chamber, cannulated on both ends with glass micropipettes, and perfused with media containing 10(6) hamster melanoma (RPMI 1856) cells/ml. In a second group of arterioles, the endothelium was denuded by running 2 ml of air through the vessel lumen. Since the tumor cells did not adhere to the vessel wall when perfused at physiologically relevant shear rates, perfusate flow was stopped and the tumor cells were allowed to settle onto the vessel wall for 20 min. After counting the number of tumor cells that settled onto the arteriolar wall, perfusate flow was re-initiated and unattached cells were washed away. The number of cells remaining adherent were counted and the percentage of adherent cells (relative to the total number of cells that settled on to the vessel wall during the period of no-flow) were calculated and compared among different groups. We observed that tumor cells are much more adhesive to denuded arterioles than to intact arterioles. To determine the mechanisms responsible for the adhesive interactions that become established and stabilized during the period of flow reduction, denuded arterioles were treated with fibronectin antiserum or Arg-Gly Asp (RGD) peptides. Both treatments significantly reduced tumor cell adhesion to denuded arterioles. In subsequent studies, melanoma cells were treated with a transglutaminase inhibitor, monodansylcadaverine (MDC), which reduced the ability of adherent tumor cells to withstand the anti-adhesive effects of a subsequent increase in perfusate flow rate after the period of no-flow. Our data suggest that tumor cells adhere to fibronectin in the subendothelial matrix in denuded arterioles by an RGD-dependent mechanism. Moreover, our observations are consistent with the concept that a transglutaminase-catalysed reaction acts to stabilize the adhesive interactions between subendothelial matrix components and melanoma cells during the period of flow stasis such that the cells are able to withstand subsequent substantial increases in wall shear rate and remain adherent. PMID- 9219732 TI - Differential influence of antiestrogens on the in vitro release of gelatinases (type IV collagenases) by invasive and non-invasive breast cancer cells. AB - Matrix metalloproteinases (MMPs) play an important role in tumor cell invasion and cancer metastasis. Accordingly, a higher level of these enzymes has been associated with the invasive phenotype. In the present study the effect of the antiestrogens, Analog II (AII), ICI-182,780 (ICI), and tamoxifen (TAM), on the in vitro release of MMPs, particularly gelatinases A and B by the MDA-MB-231 (MDA) and MCF-7 (MCF) human breast cancer cell lines was investigated using a solid phase radioassay and substrate gel zymography. Quantitatively, the enzyme activity was found to be higher in the incubation medium from estrogen receptor (ER)-negative and more metastatic MDA cells compared to ER-positive and less metastatic MCF cells. Tissue inhibitor of metalloproteinases-1 (TIMP-1) reduced the enzyme activity in media from both MDA (56.36%) and MCF (71.03%) cells. Differential antiestrogen effects on the two cell lines were observed following 4 days of treatment of cells at a concentration of 10(-6)M. The enzyme activity from MDA cells was not influenced by treatment with any of the antiestrogens, whereas, in MCF cells, ICI produced the greatest enzyme inhibition (47.93%), followed by AII (36.51%) and TAM (24.05%). Concurrent treatment of MCF cells with 17-beta-estradiol (10(-9)M) partially reversed the AII- and TAM-induced but did not alter ICI-induced inhibition of enzyme activity. Substrate gel zymography revealed that among the MMPs, the MDA cells released predominantly progelatinase A (72 kDa) along with minor bands of activated forms, 62 kDa and 59 kDa, whereas progelatinase B (92 kDa) was detected predominantly in the medium from MCF cells. Comparison of the overall antiestrogen effect indicates that ICI is the most potent inhibitor of enzyme activity in ER-positive MCF cells and that antiestrogen treatment may limit the metastatic potential of ER-positive breast cancer. PMID- 9219734 TI - Cisplatin but not BCNU inhibits urokinase-type plasminogen activator levels in human glioblastoma cell lines in vitro. AB - Glioblastomas extensively invade the surrounding normal brain tissue, with a concomitant expression of various proteolytic enzymes, in particular urokinase type plasminogen activator (uPA). In this study we used cis diamminedichloroplatinum (cisplatin) and 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), commonly used anti-cancer drugs for the treatment of glioblastomas, to study the expression of uPA in three human glioblastoma cell lines in vitro. Cells were treated with 25 microM cisplatin and 50 microM BCNU, and uPA levels were estimated by fibrin zymography during a 72-h time course. Treatment of glioblastoma cells with cisplatin resulted in significantly decreased levels of uPA in serum-free conditioned medium and cell extracts, compared to BCNU-treated and untreated cell lines. Quantitative levels of uPA enzyme activity assessed by scanning laser densitometry and uPA protein by ELISA using antibody against uPA showed decreased levels of uPA in cisplatin-treated glioma cell lines relative to BCNU and untreated cell lines. Our results suggest that anti-tumor compound, cisplatin, may exert its anti-neoplastic effects by inhibiting uPA in malignant glioblastomas. PMID- 9219733 TI - Inhibition of in vivo tumorigenicity and invasiveness of a human glioblastoma cell line transfected with antisense uPAR vectors. AB - Our previous studies showed that glioblastomas express increased urokinase-type plasminogen activator receptors (uPARs) in comparison to low-grade gliomas (Yamamoto et al., Cancer Res., 54, 5016-5020, 1994). To explore whether downregulation of uPAR inhibits tumor formation and invasiveness, a human glioblastoma cell line was transfected with a cDNA construct corresponding to 300 bp of the human uPAR's 5' end in an antisense orientation, resulting in a reduced number of uPA receptors. Co-culture studies with tumor spheroids and fetal rat brain aggregates showed that antisense SNB19-AS1 cells expressing reduced uPAR failed to invade fetal rat brain aggregates. Intracerebral injection of SNB19-AS1 stable transfectants failed to form tumors and were negative for uPAR expression in nude mice. Thus uPAR appears in this model to be essential for tumorigenicity and invasion of glioblastomas in vivo. PMID- 9219735 TI - The effect of pentoxifylline on spontaneous and experimental metastasis of the mouse Neuro2a neuroblastoma. AB - Pentoxifylline (PTX) has been reported to have both direct and indirect anti tumor effects in experimental tumor models. We studied the effect of PTX on (1) the proliferation of Neuro2a mouse neuroblastoma cells in vitro and in vivo, (2) spontaneous and experimental metastasis, (3) tumor cell membrane fluidity and (4) adhesion to a fibronectin-coated surface. PTX significantly reduced the proliferation of Neuro2a cells in vitro as determined by DNA measurement (P < 0.01) and total cell count (P < 0.02). In vivo, PTX reduced the growth of subcutaneously transplanted primary tumors in syngeneic A/J mice (P < 0.01; n = 15). All seven animals (100%) receiving intravenous tumor cells developed extensive liver metastasis. In contrast, only 1/11 (9%) of animals pre-treated with oral PTX and injected with PTX-treated cells developed liver metastases. Of five mice receiving PTX-treated cells without oral pretreatment of PTX, two out of five (40%) developed liver metastases. There was a slight, but not significant (P = 0.08) increase in both experimental and spontaneous lung metastases formation in PTX-treated animals. However, tumor nodule formation on the lung surface was inefficient. PTX also increased membrane fluidity of the Neuro2a cells and significantly decreased tumor cell adhesion to fibronectin-coated microtiter wells (P < 0.01). We conclude that PTX has a cytostatic effect on the Neuro2a mouse neuroblastoma and exerts an anti-tumor effect on liver metastases following intravenous administration of neuroblastoma cells. Whether these results are directly related to the changes in membrane properties caused by pentoxifylline remains to be established. PMID- 9219736 TI - Contemporary theories of consciousness. PMID- 9219737 TI - Frans Cornelius Donders (1818-89). PMID- 9219739 TI - British motor neuron disease twin study. AB - OBJECTIVES: To investigate the cause of sporadic motor neuron disease (MND) by twin study, so allowing (1) estimation of the genetic contribution, and (2) collection of matched pairs for a case-control study of possible environmental factors. METHODS: 10872 death certificates bearing the diagnosis MND were collected from 1979 to 1989 inclusive. Inspection of individual birth entries allowed identification of potential twins. The status of each co-twin was determined and contact made through the National Health Service Central Register (NHS-CR) and their general practitioner (GP). The diagnosis of MND was verified via the co-twin and relatives, and medical records where available. Zygosity was assessed using a recognised questionnaire. Details concerning environmental exposures and health were gathered by interview of cotwin and relatives using a semistructured questionnaire. Heritability (h2) of MND was estimated, and the environmental information was analysed by conditional logistic regression modelling. RESULTS: Seventy seven probands were identified, of whom 26 were monozygotic and 51 dizygotic. Four monozygotic probands were concordant, but two probands came from a family known to have familial MND. The estimated heritability was between 0.38 and 0.85. Most environmental risk factors were not significant. Regular vehicle maintenance (odds ratio (OR) = 7.0; 95% confidence interval (95% CI) 1.3-89.9) and occupational paint usage (OR = 3.75; 95% CI 1.0 17.1), however, occurred significantly more often in the affected cases. CONCLUSIONS: This "death discordant" method for twin collection has proved to be viable, and has allowed the ascertainment of a large population sample in a rare disease. The genetic role in sporadic MND is substantial, and higher than expected. Exposure to industrial chemicals, particularly constituents of petrochemicals and paints, may contribute to the aetiology of MND. PMID- 9219738 TI - Genetic epidemiology of multiple sclerosis. PMID- 9219740 TI - Mutilating neuropathic ulcerations in a chromosome 3q13-q22 linked Charcot-Marie Tooth disease type 2B family. AB - BACKGROUND: Charcot-Marie-Tooth disease type 2 (CMT2) or hereditary motor and sensory neuropathy type II (HMSN II) is an inherited axonal neuropathy of the peripheral nervous system. Three autosomal dominant CMT2 loci have been located on chromosomes 1p35-p36 (CMT2A), 3q13-q22 (CMT2B), and 7p14 (CMT2D) indicating that CMT2 is a genetically heterogeneous disorder. METHODS: A CMT2 family was examined for linkage to the CMT2A, CMT2B, and CMT2D loci using short tandem repeat polymorphisms. RESULTS: Suggestive evidence for linkage to 3q13-q22 was found. Recombinations occurred with markers D3S1769 and D3S1267 indicating that the CMT2B locus is located distal to D3S1267 and resides in an interval of 25 cM. Some patients in this family have pronounced sensory disturbances leading to poorly healing ulcerations. CONCLUSIONS: These unusual sensory signs for CMT were also noted in the only other CMT2B family reported so far, suggesting a distinct clinical phenotype for CMT2B. Exclusion of the locus for hereditary sensory neuropathy type I (HSN I) on chromosome 9q22 indicates that HSN I with mild motor symptoms and CMT2 with prominent sensory abnormalities are not allelic. PMID- 9219742 TI - Polyneuropathies associated with high titre antisulphatide antibodies: characteristics of patients with and without serum monoclonal proteins. AB - OBJECTIVES: Previous studies of small numbers of patients have shown that antisulphatide autoantibodies are associated with polyneuropathies having a prominent sensory component. However, clinical and electrodiagnostic features are variable. The range of clinical and electrodiagnostic findings in 19 patients with polyneuropathies and high titre (> 4500) serum IgM antisulphatide antibodies is described, together with testing for serum monoclonal (M) proteins. METHODS: About 20000 serum samples that were referred to the clinical laboratory from 1990 to the end of 1994 were screened by enzyme linked immunosorbent assay (ELISA) for specific high titre antisulphatide antibodies. The clinical and electrodiagnostic data in 23 patients with positive results were reviewed. IgM binding to peripheral nerve structures was also evaluated in these patients. RESULTS: Nineteen patients had predominantly distal, symmetric pansensory loss. Patients with IgM antisulphatide antibodies and no serum M protein usually had clinical syndromes that included: (1) neuropathic pain or dysaesthesiae, (2) no functionally significant weakness, and (3) an axonal neuropathy on electrodiagnostic testing. On immunocytochemical studies serum IgM from the patients without M proteins usually (nine of 10; 90%) bound to peripheral nerve axons, but never to myelin. Patients with antisulphatide antibodies and a serum M protein, usually IgM, were more likely than patients without a serum M protein, to have syndromes with: (1) no pain or dysaesthesiae, (2) motor abnormalities, and (3) a demyelinating polyneuropathy by electrodiagnostic criteria. In immunocytochemical studies serum IgM most often bound to either peripheral nerve myelin or endoneurial structures. CONCLUSION: Patients with polyneuropathy and high titre serum IgM antisulphatide antibodies can be classified into subgroups according to the presence or absence of a serum M protein. Patients without an M protein are more likely to have pure sensory syndromes, pain, an axonal neuropathy, and serum IgM binding to axons. Patients with a serum M protein commonly had syndromes with prominent motor involvement, no pain, and a demyelinating neuropathy. PMID- 9219741 TI - Brain and skeletal muscle bioenergetic failure in familial hypobetalipoproteinaemia. AB - OBJECTIVE: To determine whether a multisystemic bioenergetic deficit is an underlying feature of familial hypobetalipoproteinaemia. METHODS: Brain and skeletal muscle bioenergetics were studied by in vivo phosphorus MR spectroscopy (31P-MRS) in two neurologically affected members (mother and son) and in one asymptomatic member (daughter) of a kindred with familial hypobetalipoproteinaemia. Plasma concentrations of vitamin E and coenzyme Q10 (CoQ10) were also assessed. RESULTS: Brain 31P-MRS disclosed in all patients a reduced phosphocreatine (PCr) concentration whereas the calculated ADP concentration was increased. Brain phosphorylation potential was reduced in the members by about 40%. Skeletal muscle was studied at rest in the three members and during aerobic exercise and recovery in the son and daughter. Only the mother showed an impaired mitochondrial function at rest. Both son and daughter showed an increased end exercise ADP concentration whereas the rates of postexercise recovery of PCr and ADP were slow in the daughter. The rate of inorganic phosphate recovery was reduced in both cases. Plasma concentration of vitamin E and CoQ10 was below the normal range in all members. CONCLUSIONS: Structural changes in mitochondrial membranes and deficit of vitamin E together with reduced availability of CoQ10 can be responsible for the multisystemic bioenergetic deficit. Present findings suggest that CoQ10 supplementation may be important in familial hypobetalipoproteinaemia. PMID- 9219743 TI - Neurotropic viruses and Alzheimer's disease: a search for varicella zoster virus DNA by the polymerase chain reaction. AB - BACKGROUND: In studies on the possible role of viruses in the aetiopathogenesis of Alzheimer's disease, herpes simplex virus type 1 (HSV1) DNA was detected by the polymerase chain reaction (PCR) in a high proportion of normal elderly people and of patients with Alzheimer's disease. The combination of HSV1 and a host factor, the type 4 allele of the gene for apolipoprotein E, is a strong risk factor for the disease. METHODS: Brain specimens were examined for another herpes virus, varicella zoster (VZV), which, like HSV1, is neurotropic, has a predilection for residing latently in the peripheral nervous system, and can reactivate. RESULTS: Using primers for sequences in the VZV origin of replication gene or thymidine kinase gene, VZV DNA was not found in any of 24 samples (18 HSV1 positive), from 17 patients with Alzheimer's disease, nor in 20 samples (12 HSV1 positive from 12 aged normal people. Hybridisation of the PCR products with a radiolabelled oligonucleotide probe capable of detecting less than 10 copies of the target sequence, confirmed the absence of VZV DNA. CONCLUSION: The presence of one neurotropic virus--HSV1--and the absence of another--VZV--in aged human brains is consistent with a role for HSV1 in the aetiology of Alzheimer's disease. PMID- 9219744 TI - Early changes in peritumorous oedema and contralateral white matter after dexamethasone: a study using proton magnetic resonance spectroscopy. AB - AIMS: To study the mechanism of action of steroids in patients with peritumorous oedema. METHODS: To investigate early cerebral metabolic changes proton magnetic resonance spectroscopy (1H-MRS) was used before and 11 to 14 hours after treatment with dexamethasone (12 mg oral loading and 4 mg four times daily maintenance). Nine patients (two men, seven women, mean age 54) with pronounced oedema associated with various intracranial tumours (two astrocytomas, three meningiomas, two glioblastoma, and two metastases) were examined using MRI and MRS. SE1500/135 volume selected MRS (mean volume 21 ml) were performed on an oedematous region and a contralateral region. All spectra were acquired with and without water suppression. Metabolite peak area ratios were determined. RESULTS: Regions of oedema had significantly (P < 0.01) higher unsuppressed water than the contralateral regions, as expected. There was no change at this early time point after dexamethasone. The ratio of the area of choline containing compounds to that creatine and phosphocreatine compounds was determined after which the serial ratios of these before and after were calculated (a serial ratio of 1.0 would indicate no change in the choline to creatine ratios after steroid administration). The mean serial ratios for the area of oedema were 1.02 (SEM 0.08) and 1.10 (0.08) for the contralateral volume of interest, indicating no significant changes. However, significant changes (P < 0.02) were found in the N acetyl-aspartate (NAA)/choline serial ratios (0.86 (0.06) in the area of oedema, 1.20 (0.10) in contralateral brain) and the NAA/creatine serial ratios (0.86 (0.08) for the oedema, 1.25 (0.11) in contralateral brain). CONCLUSIONS: Such rapid changes may be explained either by relatively large alterations in the relaxation characteristics of NAA or, more controversially, by actual changes in the amounts of NAA. It is proposed that steroids act primarily by causing early metabolic changes that are later expressed in improvements in intracranial volume relations. PMID- 9219745 TI - Decreased cortical glucose metabolism correlates with hippocampal atrophy in Alzheimer's disease as shown by MRI and PET. AB - OBJECTIVE: To investigate the relation between atrophy of the hippocampus and parahippocampal gyrus (the % hippocampal area) and cerebral metabolic rate for glucose (CMRGlc) in Alzheimer's disease. METHODS: 13 patients with probable Alzheimer's disease by NINCDS-ADRDA criteria (six men; seven women, mean age 71 years, mini mental state 13.8 (SD 4.6)) and age matched controls were studied. T1 weighted MRI (0.5T) images were used for evaluation of the hippocampal area. With a digitiser system, a percentage of the hippocampal area to the brain (the % hippocampal area) was calculated. Eight patients received another T1 weighted MRI (1.5T) for further evaluation of the minimum thickness of the hippocampus. Regional CMRGlc (rCMRGlc) was measured using PET and the FDG technique. RESULTS: The hippocampal area in patients with Alzheimer's disease was significantly lower than that of controls (P < 0.01). All the cortical rCMRGlc values in patients with Alzheimer's disease were lower than those of controls (P < 0.01). A significant correlation (P < 0.05) was found between the % hippocampal area and rCMRGlc in the temporal lobe, temporoparieto-occipital (TPO) region, and frontal lobe in Alzheimer's disease. There was a significant correlation between minimal hippocampal thickness and ipsilateral TPO metabolism on both sides. CONCLUSION: The ipsilateral correlation between hippocampal atrophy and decreased TPO metabolism in Alzheimer's disease suggests a functional relation and the asymmetries show that Alzheimer's disease is an asymmetric disease in its early stages. PMID- 9219746 TI - Age related cognitive decline: a clinical entity? A longitudinal study of cerebral blood flow and memory performance. AB - OBJECTIVES: To evaluate the changes in regional cerebral blood flow (rCBF) and memory performance in patients with age related cognitive decline (ARCD) who did and did not become demented during a follow up period. METHODS: Twenty four patients with ARCD were recruited from an outpatient memory clinic, of whom 18 were followed up over a mean period of two years. Eighteen patients with mild to moderate probable Alzheimer's disease and 18 aged normal controls were followed up over a mean period of three years. Memory performance and rCBF were evaluated quantitatively at inclusion and during follow up, using single photon emission computed tomography with xenon-133 injection and three subtests of the Wechsler memory scale (logical memory, paired associated learning, and digit span). RESULTS: Patients with ARCD showed decreased rCBF and memory performance at initial evaluation compared with controls. Five of them became demented during the follow up period, with further decline in memory and rCBF. At inclusion, the only feature that distinguished these five patients as a group from the remainder was a pronounced temporoparietal asymmetry. The 13 patients with ARCD who did not become demented still exhibited impaired memory and rCBF at follow up, but without any further decline and no increase in flow asymmetry. CONCLUSIONS: Apart from patients in the preclinical phase of Alzheimer's disease, the ARCD category includes non-demented patients who have brain dysfunction that may represent a distinct clinical entity. PMID- 9219747 TI - The phonological loop in medicated patients with Parkinson's disease: presence of phonological similarity and word length effects. AB - OBJECTIVE: To test the verbal subsystem of Baddeley's working memory model (the phonological loop) in patients with Parkinson's disease. METHODS: Fifteen patients with idiopathic Parkinson's disease and 15 controls were tested with a span paradigm to assess the effects reflecting the functioning of the phonological loop: the phonological similarity effect (in verbal and visual presentation), and the word length effect (in visual presentation). RESULTS AND CONCLUSIONS: The patients did not show any dysfunction of the phonological loop, reflected by the presence of phonological similarity and word length effects, but had lower spans than controls. The implications of these results for the working memory model are discussed. PMID- 9219749 TI - Use of the Stroop phenomenon as a diagnostic tool for malingering. AB - AIMS: To assess a computerised version of the Stroop test for detection of malingering of cognitive deficit. METHODS: Sixty subjects were assessed using this test. Twenty had cognitive deficits due to brain damage of miscellaneous aetiologies. Ten were healthy, not acquainted with the test, and were asked to simulate cognitive impairment. Another 10 simulators were psychology students trained in the use of the test. Twenty healthy subjects served as controls. Results were analysed for reaction time, error percentage, and the Stroop effect. RESULTS: There was a significant difference in reaction time among groups, showing a direct relation of age among control subjects, and also longer reaction time in patients with brain damage than in controls. Controls and patients with brain damage showed a clear Stroop effect. Simulators had a significantly prolonged reaction time, increased error percentage, and inverted or absent Stroop effect. This alteration of the Stroop effect is never present in organic cognitive deficits and seems to be a characteristic pattern of feigning, independently of knowledge of the test. CONCLUSION: This technique is recommended as a valuable tool to detect feigned cognitive impairment. PMID- 9219748 TI - Psychiatric manifestations of neurocysticercosis: a study of 38 patients from a neurology clinic in Brazil. AB - OBJECTIVE: To determine the frequency and features of psychiatric morbidity in a cross section of 38 outpatients with neurocysticercosis. METHODS: Diagnosis of neurocysticercosis was established by CT, MRI, and CSF analysis. Psychiatric diagnoses were made by using the present state examination and the schedule for affective disorders and schizophrenia-lifetime version; cognitive state was assessed by mini mental state examination and Strub and Black's mental status examination. RESULTS: Signs of psychiatric disease and cognitive decline were found in 65.8 and 87.5% of the cases respectively. Depression was the most frequent psychiatric diagnosis (52.6%) and 14.2% of the patients were psychotic. Active disease and intracranial hypertension were associated with higher psychiatric morbidity, and previous history of mood disorders was strongly related to current depression. Other variables, such as number and type of brain lesions, severity of neuropsychological deficits, epilepsy, and use of steroids did not correlate with mental disturbances in this sample. CONCLUSIONS: Psychiatric abnormalities, particularly depression syndromes, are frequent in patients with neurocysticercosis. Although regarded as a rare cause of dementia, mild cognitive impairment may be a much more prevalent neuropsychological feature of patients with neurocysticercosis. The extent to which organic mechanisms related to brain lesions may underlie the mental changes is yet unclear, although the similar sex distribution of patients with and without depression, as well as the above mentioned correlations, provide further evidence of the part played by organic factors in the cause of these syndromes. PMID- 9219750 TI - [18F]fluorodopa PET shows striatal dopaminergic dysfunction in juvenile neuronal ceroid lipofuscinosis. AB - OBJECTIVES: To investigate whether nigrostriatal dopaminergic hypofunction is related to the extrapyramidal symptoms in patients with juvenile neuronal ceroid lipofuscinosis (JNCL). METHODS: Nine patients with JNCL and seven healthy controls were studied using [18F]fluorodopa PET. RESULTS: In the patients with JNCL [18F]fluorodopa uptake (K[i][occ]) in the putamen was 60% of the control mean and the corresponding figure in the caudate nucleus was 79%. There was a weak correlation between putamen K(i)(occ) values and extrapyramidal symptoms of the patients evaluated by the motor part of the unified Parkinson's disease rating scale (r = -0.57, P < 0.05). The overall severity of the disease also displayed a negative correlation with the K(i)(occ) values in the putamen (r = 0.71, P < 0.05). CONCLUSION: In patients with JNCL there was reduced striatal [18F]fluorodopa uptake, which had a modest correlation with extrapyramidal symptoms. Dysfunction of nigrostriatal dopaminergic neurons is therefore not the only cause of the patients' extrapyramidal symptoms, but degenerative changes in other brain areas are also contributory. PMID- 9219751 TI - The "harlequin" sign and congenital Horner's syndrome. AB - When trying to establish the likely anatomical site (preganglionic or postganglionic) of a lesion causing congenital Horner's syndrome, the distribution of facial flushing (the "harlequin" sign), may be seen. In babies and young children, facial flushing is a relatively simple clinical sign to demonstrate, compared with facial sweating. In unilateral facial flushing the areas that do not flush are almost always identical to the anhidrotic areas. However, neither facial flushing nor testing the pupil reactions with pholedrine or hydroxyamphetamine can be relied on to predict the probable site of any lesion causing congenital Horner's syndrome. Two patients with congenital Horner's syndrome are presented which demonstrated the "harlequin" sign and in whom clinical examination and pharmacological testing gave conflicting evidence for localisation of the site of the causative lesion. The presentation of congenital Horner's syndrome should be investigated and include MRI or CT to exclude a serious underlying cause. PMID- 9219752 TI - Role of the ipsilateral motor cortex in mirror movements. AB - The mechanism of mirror movements in two patients was investigated; one with congenital mirror movement, the other with schizencephaly. Transcranial magnetic stimulation on one side elicited motor evoked potentials (MEPs) in their thenar muscles on both sides with almost the same latencies, minimal thresholds, and cortical topographies. During voluntary contraction of the thenar muscle on one side, contralateral transcranial magnetic stimulation induced a silent period not only on the voluntary contraction side but on the mirror movement side and of the same duration. By contrast, ipsilateral transcranial magnetic stimulation elicited MEPs without silent periods in both muscles. With intended unilateral finger movements, an H2(15)O-PET activation study showed that the regional cerebral blood flow increased predominantly in the contralateral sensorimotor cortex, as seen in normal subjects, although mirror movements occurred. It is considered that the ipsilateral motor cortex plays a major part in the generation of mirror movements, which may be induced through the ipsilateral uncrossed corticospinal tract. PMID- 9219753 TI - SPECT, CT, and MRI in head injury: acute abnormalities followed up at six months. AB - Neuroimaging with single photon emission computed tomography (SPECT) using the cerebral blood flow tracer 99Tc(m)-HMPAO has been used to study acute functional alterations after head injury and residual abnormalities at six month follow up in 32 patients. Comparison has been made with anatomical abnormalities defined acutely with CT and on follow up with MRI. SPECT showed slightly more abnormalities than CT in the acute phase (49 regions of abnormally low tracer uptake on SPECT and 45 lesions on CT). Twenty two of the acute SPECT abnormalities were in normal regions on CT. At follow up MRI showed more abnormalities than SPECT (30 on SPECT and 48 on MRI). Ten of the SPECT deficits were in regions with normal MRI. Comparison of the intensity of late and early SPECT deficits showed that only four early deficits deteriorated whereas 28 improved. Only five of 27 lesions seen on both acute SPECT and CT resolved compared with 16 of 22 lesions seen on SPECT but not on CT. Regions of abnormally high tracer uptake were detected in the acute stage in five of the patients. No high focal uptake was evident on follow up. Ten patients with a residual SPECT abnormality and eight with residual MRI lesions were graded clinically in the upper band of good recovery. PMID- 9219754 TI - Pavlov on neuroimaging. PMID- 9219755 TI - Residual health status after Guillain-Barre syndrome. AB - To study the extent to which patients experience residual problems in daily functioning several years after having Guillain-Barre syndrome (GBS) a survey of 123 patients who had had Guillain-Barre syndrome three to six years previously was performed, using the sickness impact profile (SIP) for measuring functional health status and a functional assessment scale (F score) for measuring physical condition. The patients were diagnosed according to the international criteria for Guillain-Barre syndrome and were at the time of diagnosis unable to walk more than 10 metres without support. The physical SIP score correlated positively with final physical recovery (Pearson's r = 0.79). The psychosocial SIP score indicated impairment in all patient groups compared with matched normal control values; they included the group with no, or mild, residual symptoms (P < 0.05). No relation was found between clinical variables related to the severity or duration of Guillain-Barre syndrome and residual psychosocial dysfunctioning, except for a relation with disturbance of sensation in the arms. In conclusion, in many patients with Guillain-Barre syndrome, psychosocial functioning is still seriously affected, even when they have physically recovered, or show only mild residual signs. PMID- 9219756 TI - Cytomegalovirus infections and anti-GM2 antibodies in Guillain-Barre syndrome. AB - To investigate whether antecedent cytomegalovirus (CMV) infections in patients with Guillain-Barre syndrome are associated with the presence of specific antiganglioside antibodies, acute phase serum samples from 130 patients with Guillain-Barre syndrome and 200 controls were tested. Anti-GM2 IgM antibodies were found more often in patients with Guillain-Barre syndrome with CMV infection (22%) than in patients without the infection (2%) (P = 0.003). CMV infections may elicit anti-GM2 antibodies in susceptible patients, which may contribute to the pathogenesis of Guillain-Barre syndrome associated with CMV. PMID- 9219757 TI - Neuropsychological disorders after coronary bypass surgery. AB - OBJECTIVES: A prospective assessment of neuropsychological impairment in the early postoperative stage after coronary bypass surgery. METHODS: Seventy patients undergoing elective coronary bypass surgery (CABG) were investigated preoperatively, two to three and five to nine days postoperatively with a comprehensive neuropsychological assessment including orientation, word fluency, naming, arithmetic, memory, and visuoconstructive tasks. RESULTS: Patients exhibited significant early postoperative impairment affecting all tasks but naming. Except for the orientation measurement, most patients recovered by the fifth to ninth postoperative day. Only six patients had delirium according to DSM III-R criteria on the second or third postoperative day. Cluster analysis of neuropsychological data obtained on the second to third postoperative day identified 10 patients who were cognitively compromised. As a group, these patients had required a greater number of defibrillations and exhibited lower cardiac indices postoperatively. Preoperatively, patients at risk for postoperative dysfunction were characterised by lower verbal memory, word fluency, and clock orientation scores. CONCLUSIONS: Simple preoperative neuropsychological assessment may be helpful and clinically applicable in identifying patients at risk for postoperative cognitive dysfunction and may contribute to improve postoperative management aiming at the prevention of delirium or other transient neuropsychological disorders. PMID- 9219758 TI - Isolated angiitis of the CNS presenting as subarachnoid haemorrhage. AB - Isolated angiitis of the CNS (IACNS) commonly presents with recurrent ischaemic or haemorrhagic infarcts, but subarachnoid haemorrhage is rare. Three patients with IACNS and subarachnoid haemorrhage are reported. Florid granulomatous angiitis with Langhans and foreign body type giant cells was found at necropsy in a child with sudden death. In two other patients the diagnosis was made angiographically. In one patient multifocal infarcts on MRI became evident one week after subarachnoid haemorrhage despite initial treatment with prednisone. Subarachnoid haemorrhage may be the first presentation of IACNS. Characteristic radiographic findings may allow early diagnosis. PMID- 9219759 TI - Idiopathic intracranial hypertension in female homozygous twins. AB - The authors report on female homozygous twins with idiopathic intracranial hypertension. At the age of 12 years, both twins simultaneously developed visual disturbances with photophobia. At the age of 19 years, an ophthalmological examination disclosed papilloedema in both their eyes. At the age of 22 years, a lumbar puncture showed raised CSF pressure over (200 mm H2O) in both twins. Their neurological and radiological examinations were extremely similar; both of them had severely impaired visual acuity and impaired visual field, bilateral optic nerve atrophy, intracranial hypertension, an enlarged and partial empty sella turcica, digital markings of the calvalium, and an enlarged frontal subarachnoid space. This is the first case report describing idiopathic intracranial hypertension occurring in homozygous twins. PMID- 9219760 TI - Balo's concentric sclerosis: a clinical case study of brain MRI, biopsy, and proton magnetic resonance spectroscopic findings. AB - The antemortem diagnosis of Balo's concentric sclerosis was made in a 52 year old woman with subacute right hemiparesis on the basis of brain MRI and stereotactic brain biopsy, which showed multiple ring-like lesions of lamellated demyelination alternating with spared white matter. Proton magnetic resonance spectroscopy (1H MRS) was carried out one and nine months after the onset of illness. The first 1H MRS showed a decreased N-acetyl aspartate peak, an increased choline peak, presence of large lipid peaks, and high resonance at 1.4 ppm. The second 1H-MRS disclosed changes such as a decrease of lipid signal, a decrease of resonance at 1.4 ppm, and an increase in the myoinositol peak. These findings are similar to those reported for multiple sclerosis. It seems that this is the first report of 1H-MRS findings in Balo's concentric sclerosis. PMID- 9219761 TI - Albendazole therapy for subarachnoid cysticerci: clinical and neuroimaging analysis of 17 patients. AB - Seventeen patients with subarachnoid cysticerci received albendazole at doses of 15 mg/kg/day for eight days. All patients also received corticosteroids during the trial. Evaluation of the therapeutic response consisted of the comparison of the number of cysts shown by CT before and three months after treatment, and the evaluation of the clinical status of the patients before and after the trial. Before treatment, the 17 patients had 30 subarachnoid cysticerci, 11 of which were > 50 mm in diameter. Seventeen cysts were located at the convexity of cerebral hemispheres, seven at the sylvian fissure, five at the ambiens cisterns, and one at the cerebellopontine angle cistern. Fourteen patients had seizures, 10 had hemiparesis, three were demented, one had diminution of visual acuity, and one had hemifacial spasm. Brain CT obtained after therapy showed resolution of 27 cysts (90% effectiveness). Fourteen (82%) patients had total resolution of all cysts. All but three patients were asymptomatic. Remaining deficits included hemiparesis in two patients and diminution of visual acuity in one. It is concluded that albendazole is an effective treatment for subarachnoid cysticerci as it causes disappearance of most lesions on CT, and produces considerable improvement in the clinical manifestations of the patients. PMID- 9219762 TI - Saccade velocity in idiopathic and autosomal dominant cerebellar ataxia. AB - Slow saccades are often found in degenerative ataxia. Experimental studies have shown that horizontal saccades are generated in the paramedian pontine reticular formation and that lesions in this area produce slow saccades. Based on these findings, saccade slowing should be a frequent feature of olivopontocerebellar atrophy, a type of cerebellar degeneration with prominent involvement of the pons. To test this hypothesis, saccade velocity was measured in 31 patients with autosomal dominant cerebellar ataxia (ADCA) and 17 patients with idiopathic cerebellar ataxia (IDCA). Saccade velocity was reduced in most patients with ADCA whereas it was normal in IDCA although olivopontocerebellar atrophy occurred in both groups. Saccade velocities correlated with pontine size in ADCA but not in IDCA. The data disprove the hypothesis that saccadic slowing is a clinical hallmark of olivopontocerebellar atrophy. Instead, only patients with ADCA and morphological features of olivopontocerebellar atrophy have slow saccades. PMID- 9219763 TI - Altered pupillary size and darkness and light reflexes in Alzheimer's disease. AB - The purpose was to compare resting pupil diameter in darkness and light, and the pupillary darkness and light reflexes between a group of patients with Alzheimer's disease and a group of healthy old people. Nine medication free patients with Alzheimer's disease and nine healthy control subjects, matched for sex and age with the patients, participated. There were six men and three women and the median age was 72 years in both groups. Pupil diameter was monitored with an infrared television pupillometer. Resting pupil diameter was smaller in the Alzheimer's disease group (P = 0.041, in darkness). The amplitude and the maximum dilatation velocity of the darkness reflex were smaller for the Alzheimer's disease group (maximum dilatation velocity P < 0.002). The amplitude and the 75% recovery time of the light reflex response were reduced in the Alzheimer's disease group (P < 0.002 and P = 0.034 respectively). There was no difference in the latency of the reflex response between the two groups. The reduced pupil size and diminished darkness reflex in the Alzheimer's disease group are consistent with a sympathetic deficit in the patients. The reduction in light reflex amplitude and recovery time are likely to be secondary to the grossly diminished pupil size in the patient group. The lack of any change in light reflex latency in the patients with Alzheimer's disease argues against an afferent defect. The pupillary changes in patients with Alzheimer's disease are qualitatively the same as those seen in healthy old people and are consistent with the notion of "accelerated aging" in Alzheimer's disease. PMID- 9219764 TI - Recurrent neck pain as a variant of migraine: description of four cases. AB - Four patients who had recurrent attacks of idiopathic unilateral neck pain and tenderness of the ipsilateral carotid artery are described. Two patients had never had headache. The other two had migraine without aura. All patients had dilatation of extracranial arteries during the attacks (telethermographic examination), oculosympathetic hypofunction (pupillary tests), and positive responses to vasoactive drugs which are commonly used for migraine treatment. Recurrent neck pain involving the carotid artery seems to be a variant form of migraine that may occur alone or in association with headache in patients with involvement of extracranial arteries. PMID- 9219765 TI - The thermolabile variant of 5,10-methylenetetrahydrofolate reductase is not associated with Parkinson's disease. PMID- 9219767 TI - Contraversive visual tilt illusion associated with a cerebellar infarction. PMID- 9219766 TI - Refsum's disease: long term treatment preserves sensory nerve action potentials and motor function. PMID- 9219768 TI - Low striatal D2 receptor binding as assessed by [123I]IBZM SPECT in patients with writer's cramp. PMID- 9219769 TI - Chronic sensory ataxic neuropathy and ophthalmoplegia with oculomotor nerve hypertrophy associated with IgM antibodies against gangliosides containing disialosyl groups. PMID- 9219770 TI - Proceedings of the winter meeting of the British Neuropsychiatry Association, London Zoo, Regent's Park, London, 17 January 1997. PMID- 9219771 TI - Clinical overdiagnosis of vascular dementia versus necropsy confirmed series. PMID- 9219772 TI - Delayed gastric emptying in reflux patients: to be or not to be? PMID- 9219773 TI - Cutting edge management of achalasia. PMID- 9219774 TI - Hereditary pancreatitis. PMID- 9219775 TI - Diagnosis and treatment of gastrointestinal bleeding secondary to portal hypertension. American College of Gastroenterology Practice Parameters Committee. AB - Guidelines for clinical practice are intended to suggest preferable approaches to particular medical problems as established by interpretation and collation of scientifically valid research, derived from extensive review of published literature. When data are not available that will withstand objective scrutiny, a recommendation may be made based on a consensus of experts. Guidelines are intended to apply to the clinical situation for all physicians without regard to specialty. Guidelines are intended to be flexible, not necessarily indicating the only acceptable approach, and should be distinguished from standards of care, which are inflexible and rarely violated. Given the wide range of choices in any health care problem, the physician should select the course best suited to the individual patient and the clinical situation presented. These guidelines are developed under the auspices of the American College of Gastroenterology and its practice parameters committee. These guidelines are also approved by the governing boards of American College of Gastroenterology and Practice Parameters Committee. Expert opinion is solicited from the outset for the document. Guidelines are reviewed in depth by the committee, with participation from experienced clinicians and others in related fields. The final recommendations are based on the data available at the time of the production of the document and may be updated with pertinent scientific developments at a later time. The following guidelines are intended for adults and not for pediatric patients. OBJECTIVE: To develop practice guidelines for the management of gastrointestinal bleeding in adult patients with cirrhosis and portal hypertension. METHOD: Randomized controlled trials published through October of 1993 were evaluated by members of the American College of Gastroenterology Practice Parameters Committee. Each paper was reviewed by three members of the committee and rated for quality of design by predetermined criteria. Meta-analysis of the studies for each treatment were evaluated for both outcome and quality of design and formed the basis for recommendations for treatment. Randomized controlled trials published between October of 1993 and August of 1995 have been added to update and modify the recommendations. The reader is referred to an excellent article by D'Amico et al. (The treatment of portal hypertension: A meta-analytic review. Hepatology 1995;22:332-354), which presents most of the meta-analyses reviewed by this committee. CONCLUSIONS: Once esophageal varices have been established by endoscopy as the site of bleeding, either sclerotherapy or endoscopic variceal ligation should be performed to control the bleeding episodes. Concomitant use of vasoactive drugs lowers portal pressure, potentially offers the endoscopist a clearer field in which to work, and is the only noninvasive treatment for nonesophagogastric variceal sites of bleeding related to portal hypertension. For patients failing medical therapy, the transjugular intrahepatic portasystemic shunt procedure is a reasonable alternative to an emergency surgically created shunt. Nonselective beta-adrenergic blockers are the only proven therapy for prevention of first variceal hemorrhage. Both nonselective beta-adrenergic blockers and endoscopic variceal ligation (which has replaced sclerotherapy for this indication) are effective in reducing the risk of recurrent variceal bleeding. For patients failing these approaches, selective or total shunts or, in selected patients, liver transplantation are appropriate rescue procedures. PMID- 9219776 TI - Giant diverticula of the colon: a clinical assessment. AB - Giant colonic diverticulum is a rare complication of diverticular disease. In the English literature, only 81 cases have been described. Twelve patients had complications caused by the giant diverticulum. Seventy patients were treated operatively, and three died. Elective resection of the diverticulum and the adjacent colon with primary anastomosis is the ideal treatment. The significant number of complications caused by the giant diverticulum and the low morbidity and mortality rate associated with surgical treatment reinforce the importance of accurate diagnosis and elective treatment of this disorder. PMID- 9219777 TI - Helicobacter pylori and acid peptic disorders of the esophagus: is it conceivable? PMID- 9219778 TI - Gastric electrical dysrhythmias and delayed gastric emptying in gastroesophageal reflux disease. AB - OBJECTIVE: Deranged gastric motility and delayed gastric emptying are commonly implicated in the pathophysiology of gastroesophageal reflux disease. We measured gastric electrical activity and gastric emptying time of a solid-liquid meal by electrogastrography and antral ultrasound, respectively, in 42 patients with gastroesophageal reflux disease (age 7.4 +/- 1.6 yr). METHODS: Based on endoscopy and histology of the esophageal mucosa, reflux disease was moderate in 20 patients and severe in 22. Electrogastrography was measured by placing two Ag AgCl electrodes on the epigastric skin, signals were digitized and fed into a personal computer, and data were obtained by running spectrum analysis. The electrogastrographic variables calculated were: 1) percent of electrical dysrhythmias and normal electrical rhythm (bradygastria or 0.5-2.0 cycles/min, tachygastria or 4.0-9.0 cycles/min; normal rhythm is 2.0-4.0 cycles/min); 2) fed:fasting ratio of dominant electrogastrographic power; 3) fed:fasting ratio of the dominant frequency instability coefficient. Gastric emptying time and electrical activity results were compared with those measured in 15 controls (7.1 +/- 1.7 yr). RESULTS: Dysrhythmic episodes were more common in both groups of patients than in controls (p < 0.01); furthermore, gastric emptying time was significantly more delayed in patients than in controls (p < 0.01). Children with severe gastroesophageal reflux were distinguished from those with moderate disease for post-feeding gastric electrical abnormalities consisting of reduced electrogastrographic dominant power and increased frequency variability (p < 0.01), as well as for a more prolonged gastric emptying time (p < 0.05). Prevalence of both normal electrical rhythm and dysrhythmias did not discriminate the two groups of patients. In patients and in controls, a significant inverse correlation between fed electrogastrographic power and gastric emptying time was found (r -0.88, p < 0.01). CONCLUSIONS: Fed gastric electrical abnormalities consisting of reduced dominant power and increased variability of the electrical dominant frequency are detected in patients with severe gastroesophageal reflux disease and are associated with delayed gastric emptying. Gastric electrical dysrhythmias may be included among the pathogenetic components of gastroesophageal reflux disease. PMID- 9219779 TI - A prospective assessment of gastroesophageal reflux before and after treatment of achalasia patients: pneumatic dilation versus transthoracic limited myotomy. AB - OBJECTIVES: We conducted this study to determine whether reflux should be a major consideration in the choice of treatment for achalasia patients. Achalasia patients undergoing either pneumatic dilation or transthoracic limited esophagomyotomy were monitored for reflux before and after treatment, for comparison. METHODS: Twenty-four hour ambulatory esophageal pH tests and esophageal manometry were performed on 32 consecutive, untreated achalasia patients. Studied (before and after treatment) were 17 patients who underwent pneumatic dilation and 15 patients who received transthoracic limited myotomy without fundoplication. All follow-up studies were completed within 12 months of treatment. RESULTS: The ages of the two groups were not significantly different (p > 0.05, 45 +/- 9 yr myotomy vs. 44 +/- 13 yr dilation). The resting lower esophageal sphincter pressure was not significantly different (p > 0.05 before treatment) between groups but was reduced significantly (p < 0.05 after treatment) in both groups (30 +/- 9 mm Hg before vs. 9 +/- 4 mm Hg after myotomy, and 27 +/- 10 mm Hg before vs. 11 +/- 4 mm Hg after pneumatic dilation. The total time the pH was < 4.0 was not significantly different, p > 0.05, in either group before treatment (myotomy, 3.7 +/- 4.4%; dilation, 2.9 +/- 4.9%) or after treatment (myotomy, 8.6 +/- 9.2%; dilation, 10.2 +/- 15.9%). Twelve of 32 patients (38%), had a percent total time < 4.0 that exceeded 6% after treatment, eight of whom were asymptomatic. CONCLUSIONS: These results indicate that the amount of reflux after treatment by both pneumatic dilation and transthoracic esophagomyotomy is similar. The absence of reflux symptoms in treated achalasia patients does not exclude the possibility of significant acid reflux. PMID- 9219780 TI - Clinical characteristics of hereditary pancreatitis in a large family, based on high-risk haplotype. The Midwest Multicenter Pancreatic Study Group (MMPSG) AB - OBJECTIVES: Because there are no markers for hereditary pancreatitis (HP), diagnosis has relied on clinical features and inferences. Identification of the HP disease gene locus on chromosome 7q35 provides the first genetic marker for HP, allowing an accurate comparison of the clinical diagnosis of HP with the presence of a high-risk HP haplotype. Our objectives were to compare the clinical diagnosis of HP with inheritance of the HP gene and to characterize the common clinical features. METHODS: A detailed questionnaire was administered to 102 study participants of a large HP kindred. Blood samples were taken for DNA extraction and high-risk haplotype determination. Clinical findings were compared with the presence of a high-risk haplotype. RESULTS: A family tree of more than 500 members and eight generations was constructed, and clinical features of the 102 participants were determined. HP occurred before the age of 5 yr in 58% of subjects, who presented with common symptoms of abdominal pain, nausea/vomiting, and frequent attacks. Thirty-five probands, of whom 80% had clinical symptoms, carried the high-risk haplotype, confirming previous estimates of 80% penetrance. Thirty-two of the study participants had been clinically diagnosed with HP, whereas 70 were clinically unaffected. With regard to the presence of the high risk haplotype, 87.5% of the clinically diagnosed patients were affected by HP (true positive), whereas 12.5% did not carry the high-risk haplotype (false positive). Seven obligate carriers were identified through DNA analysis; three had previously been unrecognized because of lack of affected offspring. CONCLUSIONS: The diagnosis of hereditary pancreatitis on clinical grounds alone may be inaccurate in less severe cases, as is the exclusion of carrier status through family tree analysis. Therefore, a definitive diagnosis of hereditary pancreatitis in equivocal cases or exclusion of a carrier state should include analysis of genetic markers. PMID- 9219781 TI - Lack of spontaneous regression of tubular adenomas in two years of follow-up. AB - OBJECTIVE: Change in colon polyp size over time has not been well characterized. It has been inferred that some polyps will increase in size, leading to an increased risk of progressing to cancer, whereas other polyps may spontaneously regress. To develop a better understanding of the natural history of colon polyps, we prospectively investigated change in polyp size over a 2-yr period. METHODS: Patients were enrolled if they had an endoscopically detected proximal rectal or sigmoid polyp measuring 3-9 mm. The index polyp site was then permanently marked with an adjacent India ink tattoo to allow definitive future localization of the polyp. Patients underwent flexible sigmoidoscopy at 6-month intervals, and at each examination, the polyp size was carefully measured with open biopsy forceps. After a maximum of 2 yr, each polyp was removed and the histology determined. RESULTS: Thirty polyps were followed in 26 patients who completed the study. Twelve polyps were tubular adenomas (TA), one was tubulovillous, 14 were hyperplastic polyps (HP), two had no pathological diagnosis, and one was a leiomyoma. HP were more likely to decrease in size than were TA. Three polyps demonstrated fast growth rates (2-4 mm/yr), and all were TA. Two polyps were removed early because their size had reached 1 cm or more. Both of those polyps were TA. No polyps regressed completely during the 2 yr of the study; neither did we find consistent linear growth rates. CONCLUSIONS: In contrast to prior reports, in this study, after polyps had been definitively localized with India ink, we observed no complete polyp regressions. PMID- 9219782 TI - Prospective randomized comparison of stonetome combination catheter versus conventional endoscopic clearance of common bile duct. AB - OBJECTIVES: To assess the therapeutic efficacy and performance characteristics of a novel cannula that has recently become available for clearance of the common bile duct (CBD), combining a sphincterotome and retrieval balloon in a single instrument, and to compare it with the conventional approach. METHODS: Seventeen consecutive patients with signs of biliary obstruction were prospectively randomized to undergo evaluation and clearance of the CBD by combination catheter (Stonetome, Microvasive) or conventional sphincterotomy (Ultratome) with separate balloon sweep. In all cases, three sweeps of the CBD were made with an 11.5-mm retrieval balloon after sphincterotomy. Fluoroscopy time was measured, as was total time from initiation of sphincterotomy to removal of endoscope from the patient. The same endoscopist performed all procedures. RESULTS: Patient groups were matched with regard to age, gender, proportion with prior cholecystectomy, CBD diameter, and stone size. Patients were successfully treated by either approach. Use of the Stonetome combination catheter reduced mean fluoroscopy time by 52% (from 3.1 +/- 0.6 to 1.5 +/- 0.2 min), mean total time by 57% (from 11.3 +/- 2.2 to 4.9 +/- 1.1 min), and dosage of midazolam used for conscious sedation. CONCLUSIONS: The new combination catheter allows the elimination of one step in exchanging the sphincterotome for a retrieval balloon after sphincterotomy. Its efficacy in clearing the CBD is equal to that of our conventional approach. Use of the combined approach should minimize the risk of losing cannulation, reduce fluoroscopic exposure, and improve the overall efficiency of the procedure. PMID- 9219783 TI - Sphincter of Oddi dysfunction is associated with chronic pancreatitis. AB - OBJECTIVES: Chronic pancreatitis can be caused by ductal obstruction resulting from cicatricial papillary stenosis, but sphincter of Oddi motility studies have been inconclusive in patients with established chronic pancreatitis. We sought to determine whether there is an association between papillary sphincter dysfunction and changes of early chronic pancreatitis. METHODS: Consecutive patients who underwent sphincter of Oddi manometry to investigate unexplained upper abdominal pain (n = 104) were assessed for evidence of chronic pancreatitis by pancreatic ductography, endoscopic ultrasound, and pancreatic fluid bicarbonate concentration. RESULTS: Sphincter of Oddi dysfunction patients were four times more likely (odds ratio = 4.6) to have evidence of chronic pancreatitis than were those without sphincter dysfunction (p = 0.01). Of 68 patients with sphincter of Oddi dysfunction, 20 (29%) had structural evidence of chronic pancreatitis. Twenty of 23 (87%) patients with chronic pancreatitis had sphincter of Oddi dysfunction. CONCLUSIONS: Sphincter of Oddi dysfunction is associated with changes of chronic pancreatitis in patients with unexplained pancreaticobiliary pain. Longitudinal follow-up is required to confirm these preliminary findings. PMID- 9219784 TI - Influence of hepatitis C virus genotypes and HIV infection on histological severity of chronic hepatitis C. The Hepatitis/HIV Spanish Study Group. AB - OBJECTIVES: The factors influencing the histological severity of chronic hepatitis C (CHC) have not been well established. We therefore investigated the effect of hepatitis C virus (HCV) genotypes and human immunodeficiency virus (HIV) infection on histological liver damage in a cohort of intravenous drug users with CHC. METHODS: We analyzed the histological activity score and the HCV genotypes in 59 HCV-RNA-positive patients with biopsy-proven CHC. Forty-eight (81%) of them had concomitant HIV infection with a CD4+ cell count above 200 x 10(6) cells/L and an absence of AIDS-defining conditions. Multivariate analysis was performed to determine the features associated with the histological severity. RESULTS: Minimal/mild hepatitis was found in 16 patients (27%), moderate chronic hepatitis in 29 (49%), and severe chronic hepatitis in 14 (24%). Patients with HCV subtype 1b had a higher histological score than others (8.7 +/- 3.3 vs. 6.5 +/- 3.2, p = 0.012), either as single or mixed infections. In multivariate analysis, HIV-infected individuals had a higher score of piecemeal necrosis (OR = 21.7, p = 0.002) and a higher stage of fibrosis (OR = 17.9, p = 0.004) than patients without HIV infection. HIV infection and HCV genotype 1b were found to be independent factors of histological severity. CONCLUSIONS: Liver damage in patients with CHC seems to be directly influenced by HCV subtypes. Infection by HCV subtype 1b is closely associated with more severe forms of liver pathology. Furthermore, the presence of HIV infection is an independent factor associated with more aggressive histological damage. In these patients, higher degrees of piecemeal necrosis and fibrosis are commonly seen. PMID- 9219785 TI - Detection of serum antibodies to Helicobacter pylori by an immunochromatographic method. AB - OBJECTIVES: FlexsureHP is a bi-directional immunochromatographic device for detection of IgG antibodies to Helicobacter pylori in human serum. This test, requiring only three steps and a 4-min incubation, can be used as an office-based diagnostic test. The goal of this study was to compare the sensitivity and specificity of the FlexsureHP, when performed by a clinician, with the established ELISA in the evaluation of clinical samples in an office setting. RESULTS: The sensitivity and specificity of the FlexsureHP, compared with the biopsy, is 92.4% and 83.0%, respectively, with a positive predictive value of 88.4%. This was not significantly different from the results obtained when the ELISA was compared with biopsy data. CONCLUSION: The immunochromatographic device, FlexsureHP, is a rapid, highly sensitive, and moderately specific office based diagnostic test for H. pylori infection. PMID- 9219786 TI - Eradication of H. pylori in a developing country: comparison of lansoprazole versus omeprazole with norfloxacin, in a dual-therapy study. AB - OBJECTIVES: Lansoprazole, a newer benzimidazole, is more potent than omeprazole in its anti-Helicobacter pylori effect in vitro. The present study was aimed at assessing its efficacy in a developing country. METHODS: Fifty patients were randomized to receive either lansoprazole or omeprazole, with norfloxacin for 2 wk; the ulcer healing rates, H. pylori eradication rates, and recurrence rates were compared over a 6-month period. RESULTS: Both lansoprazole and omeprazole were equally effective in inducing healing of ulcer (96.1 vs 95.5%, p > 0.05) and eradicating H. pylori (76.9 vs 63.9%, p > 0.05), with very low recurrence rates over a 6-month follow-up period. CONCLUSIONS: Either lansoprazole or omeprazole combined with norfloxacin is effective in eradicating H. pylori in a high percentage of cases of duodenal ulcer, with little difference between the two proton pump inhibitors. PMID- 9219787 TI - Combined therapy with 5-aminosalicylic acid tablets and enemas for maintaining remission in ulcerative colitis: a randomized double-blind study. AB - OBJECTIVES: To assess the efficacy of a combination of oral and topical 5 aminosalicylic acid (5-ASA) for the maintenance treatment of ulcerative colitis, we undertook a double-blind randomized clinical trial. METHODS: Patients aged 18 to 65 yr (with disease extent greater than proctitis only) were eligible for inclusion in the study if they met the following criteria: (a) history of two or more relapses in the last year; (b) achievement of remission in the last 3 months (with maintenance of remission for at least 1 month). Patients enrolled in the study were randomly assigned to one of the two following 1-yr treatments: (1) combined therapy with 5-ASA tablets 1.6 g/day and 5-ASA enemas 4 g/100 ml twice weekly; (2) oral therapy with 5-ASA tablets 1.6 g/day and placebo enemas/twice weekly. The main end point of the study was the maintenance of remission at 12 months. RESULTS: Upon completion of the study, relapse occurred in 13 of 33 patients in the combined treatment group versus 23 of 36 patients in the oral treatment group (39 vs 69%; p = 0.036). No significant side effects related to treatment were observed in either group. A simplified pharmacoeconomic analysis shows that this form of combined treatment can have a favorable cost effectiveness ratio. CONCLUSIONS: Our results indicate that 5-ASA given daily by oral route and intermittently by topical route can be more effective than oral therapy alone. This form of combination treatment can be appropriate for patients at high risk of relapse. PMID- 9219788 TI - Increased prevalence of lactose malabsorption in Crohn's disease patients at low risk for lactose malabsorption based on ethnic origin. AB - OBJECTIVES: The aim of this study was to compare the prevalence of lactose malabsorption (LM) in various subgroups of inflammatory bowel disease patients with controls matched for age, sex, and ethnic origin. METHODS: In 260 patients with IBD [121 Crohn's disease (CD) and 139 ulcerative colitis (UC)] and 158 controls at low and moderate risk for LM the prevalence of lactose malabsorption was determined by H2 breath testing. RESULTS: A control group at low ethnic risk had a prevalence of LM of 29.2% compared with 40.0% in CD (p < 0.025) and 13.3% of ulcerative colitis patients (p < 0.025). No significant differences were observed in comparable groups at moderate risk for LM. Irrespective of ethnic origin, 68.1% of patients with CD limited to the terminal ileum were lactose malabsorbers compared with 43.5% of patients with Crohn's colitis (p < 0.05). Additional analysis according to anatomical location indicated that Crohn's disease of the proximal small bowel (duodenum, jejunum), terminal ileum, terminal ileum plus colon, and colon alone were associated with a prevalence of LM of 100, 68.1, 54.5, and 43.5% respectively. CONCLUSIONS: In patients at low ethnic risk there is a statistically significant increase in the prevalence of LM in CD patients and a decreased prevalence in ulcerative colitis compared with controls. PMID- 9219789 TI - Pulmonary function abnormalities in patients with ulcerative colitis. AB - OBJECTIVES: We hypothesized that abnormalities in pulmonary function are present in subjects with ulcerative colitis (UC). Our primary objectives were (1) to determine the frequency of pulmonary abnormalities in a sample of subjects with UC, and (2) to compare UC subjects with abnormal pulmonary function tests (PFT) with those who have normal PFT in terms of smoking behavior and history of preexisting pulmonary disease. METHODS: We selected 100 consecutive UC subjects from the index of subjects currently attending the University of Calgary clinics. Sixty-six were eligible. Fifty-five (83%) agreed to participate. RESULTS: PFT abnormalities were found in 30 (55%) subjects. Abnormal PFT could not be predicted by current or past smoking status, family history of respiratory diseases, occupational history, or current medication status. CONCLUSIONS: (1) PFT abnormalities were detected in half of the subjects. (2) Smoking status was not predictive of these abnormalities. PMID- 9219790 TI - Altered bone metabolism in inflammatory bowel disease. AB - A reduced bone mineral density has been reported in inflammatory bowel disease (IBD). OBJECTIVE: To assess the mechanisms of bone disease in IBD. METHODS: We studied in 90 patients (61 with Crohn's disease, 22 with ulcerative colitis, 7 with indeterminate colitis) biochemical markers of bone metabolism in serum and bone mineral density by peripheral quantitative computed tomography at the forearm. RESULTS: Forty-five percent of the patients had a reduced bone density (Z score < -1). Serum calcium was normal in most patients, vitamin D deficiency was documented in 17%. Osteocalcin, a serum marker of bone formation, was decreased in 26% (1.2 +/- 0.1 ng/ml), whereas the carboxyterminal cross-linked telopeptide of type I collagen (ICTP), a recently described serum parameter of bone breakdown, was stimulated in 38% (10.4 +/- 2.3 microg/L). Of 33 patients with increased ICTP levels, 19 showed a decreased bone density (Z score < -1), and 2 of them never received steroids. An active status of the underlying disease in most patients with increased ICTP levels suggests a direct effect of the underlying IBD. In the whole series of patients with a history of active disease (n = 34), 47% had signs of an increased bone degradation (ICTP > 5 microg/L; mean, 12.9 +/- 4.7 microg/L). Data derived from a retrospective survey of 245 patients with IBD suggest that the prevalence of bone fractures in IBD is unexpectedly high, particularly in patients with a long duration of disease, frequent active phases, and high cumulative doses of corticosteroid intake. CONCLUSIONS: Several mechanisms may be involved in IBD-associated bone disease: (1) a high inflammatory activity directly induces bone degradation via yet unknown pathways, (2) treatment with corticosteroids may exert catabolic effects on the bone, or (3) malabsorption and vitamin D deficiency may activate bone turnover. PMID- 9219791 TI - Significance of interleukin-1beta and interleukin-1 receptor antagonist genetic polymorphism in inflammatory bowel diseases. AB - OBJECTIVE: Genetic susceptibility to inflammatory bowel disease is well recognized. There is also increasing evidence for the activation of the mucosal immune system and the production of inflammatory cytokines, i.e., interleukin (IL)-1ra and IL-1beta in the inflammatory bowel disease. The aim of this study was to analyze the IL-1beta and IL-1ra gene polymorphism and linkage disequilibrium coefficient between the different alleles of these genes in patients with Crohn's disease (CD) or ulcerative colitis (UC), according to the severity of the disease. METHODS: Two hundred twenty-eight inflammatory bowel disease patients (87 UC and 141 CD) were included in this study and compared with 113 unrelated controls. The IL-1beta and IL-1ra gene polymorphism was studied after specific amplification of variable regions by PCR. A penta-allelic polymorphism, corresponding to a VNTR region located in intron 2 of the IL-1ra gene, was analyzed, whereas bi-allelic RFLPs displayed by two restriction enzymes (TaqI and AvaI) at position -511 of the IL-1beta gene were analyzed. RESULTS: There was no significant difference of genotype distribution between controls and CD or UC patients. However, surgically treated UC patients were characterized by a higher frequency of genotype IL-1ra 1-2 (39 vs 16%, pc < 0.01) compared with nonoperated UC patients. Moreover, nonoperated UC patients displayed a lower frequency of IL-1ra allele 2 than surgically treated UC patients (14 vs 34%, pc < 0.002) or controls (14 vs 30%, pc < 0.005). Furthermore, simultaneous analysis of the IL-1beta and IL-1ra genes that are located in the same region of chromosome 2 revealed that CD patients carrying the IL-1beta allele 2 were more often noncarriers of IL-1ra allele 2 (p < 0.005). Moreover, UC and CD patients were, characterized by a lower frequency of the association of IL-1ra allele 2 and IL 1beta allele 2 compared with controls (8.3 vs 20.3% and 10.6 vs 20.3%, p < 0.03). CONCLUSIONS: IL-1ra and IL-1beta gene polymorphism analysis from a clinical standpoint might help in defining UC prognosis. However, functional studies at both the circulating and mucosal level with stratification on allele associations, especially IL-1ra allele 2-IL-1beta allele 2 subgroups must be realized before therapeutic implications. PMID- 9219792 TI - Expression of cyclooxygenase-2 mRNA in active inflammatory bowel disease. AB - OBJECTIVES: In inflammatory bowel disease (IBD), increased amounts of prostaglandins correlate to disease activity. Prostaglandins are produced via the cyclooxygenase (COX) pathway and exhibit both pro- and anti-inflammatory effects. Whereas COX-1 is a constitutive enzyme present at all times and is thought to produce the cytoprotective prostaglandins, COX-2 represents the inducible form of cyclooxygenase leading to production of proinflammatory prostaglandins. In inflammatory bowel disease it is yet unclear whether COX-2 plays a role in the inflammatory response. The purpose of this study was to evaluate the role of COX 2 in inflammatory bowel disease. METHODS: Of the 44 individuals included in the study, 22 had ulcerative colitis, 11 had Crohn's disease, and 11 were healthy controls. Standard rigid rectoscopy was performed. The degree of inflammation was assessed using a semiquantitative scale. A biopsy was taken from the most affected area. mRNAs for COX-1 and COX-2 were detected using reverse transcription-polymerase chain reaction. RESULTS: The fraction of patients demonstrating COX-2 mRNA significantly increased with increasing disease activity (p < 0.005), whereas the fraction of patients demonstrating COX-1 mRNA remained unchanged (p > 0.05). CONCLUSIONS: This study demonstrates a clear relationship between endoscopic activity and relative presence of mRNA for COX-2. In contrast mRNA for COX-1 is detected equally often. This indicates that COX-2 is involved in the acute inflammatory response of chronic inflammatory bowel disease. PMID- 9219793 TI - Elevated serum CA 125 concentration in patients with tuberculous peritonitis: a case-control study. AB - OBJECTIVES: High serum cancer antigen (CA) 125 levels have been shown to be present in patients with ovarian carcinoma, nongynecological cancers, and some benign diseases and have been used as a useful marker for monitoring patients with epithelial ovarian cancer. Although tuberculous peritonitis with elevated serum CA 125 levels is an important diagnostic problem because it may lead to misdiagnosis as ovarian carcinoma and unnecessary laparatomies, the levels of CA 125 in patients with tuberculous peritonitis has not been studied in detail. METHODS: Serum CA 125 levels in 11 consecutive patients (2 males, 9 females) with tuberculous peritonitis admitted to Hacettepe University and 20 healthy adult controls (7 males, 13 females) were studied. RESULTS: Serum CA 125 levels were found to be elevated in all patients with tuberculous peritonitis. The mean level in the study group was 316.6 IU/ml, whereas the level was 13.8 IU/ml in the control group (p < 0.0001). Serum CA 125 normalization showed a very close correlation with the response to antituberculous therapy. CONCLUSIONS: Tuberculous peritonitis must be considered in a differential diagnosis of patients with elevated serum CA 125 concentration. Correct diagnosis may prevent unnecessary laparatomies performed under the assumption of ovarian carcinoma, because it is cured completely by antituberculous therapy. Serum CA 125 level might be used as an effective marker in the diagnosis and follow-up of patients with tuberculous peritonitis. PMID- 9219794 TI - Endothelin-1 in the gastric mucosa in stress ulcers of critically ill patients. AB - OBJECTIVE: Gastric microcirculatory disturbances are involved in the pathogenesis of stress ulcers; however, vasomodulators causing this process are not fully understood. This study was conducted to investigate the role of endothelin 1 (ET 1), a potent vasoconstrictive peptide, in stress ulcers in critically ill patients. METHODS: Using sandwich enzyme immunoassay, we measured ET-1 content in plasma and the gastric mucosa of 16 critically ill patients with traumatic head injury on admission and of 11 healthy subjects. Gastric mucosal samples were obtained endoscopically. When gastric drainage contained occult blood, endoscopic examination was performed again, and ET-1 concentrations in injured and adjacent normal mucosa were compared. RESULTS: Plasma and mucosal ET-1 concentrations were significantly higher in critically ill patients on admission (6.1 +/- 0.6 pg/ml and 13.8 +/- 1.6 ng/g, respectively) compared with values in control subjects (2.7 +/- 0.4 pg/ml and 8.2 +/- 0.5 ng/g, respectively) (p < 0.01). The mucosal ET 1 concentration tended to be elevated in patients who had experienced hypoxia compared with those who had not (p = 0.07). In five patients who were again examined endoscopically, the ET-1 concentration in the injured mucosa was significantly higher than that in adjacent mucosa (19.2 +/- 3.2 and 10.1 +/- 1.6 ng/g, respectively; p < 0.05). CONCLUSIONS: These results suggest that endogenous ET-1 plays an important role in the local pathogenesis of stress ulcers, especially those caused by hypoxia. PMID- 9219795 TI - The prediction of lymph node metastases in colorectal cancer by expression of the nucleoside diphosphate kinase/nm23-H1 and histopathological variables. AB - OBJECTIVE: To ascertain the risk of locoregional lymph node metastases from colorectal cancer, we compared microscopic pathological characteristics of the primary tumor with the expression of the nm23-H1 protein. METHODS: The nm23-H1 expression of 100 colorectal carcinomas and corresponding non-neoplastic mucosa was analyzed immunohistochemically at the time of primary curative surgery (R0 resection). Conventional histopathological factors (depth of infiltration, grade of differentiation, invasion of lymph vessels or veins) that are proven indicators for metastatic involvement of locoregional lymph nodes were examined in all cases. RESULTS: Of 45 tumors with lymph node metastases, 42 (93%) had a low nm23-H1 expression whereas only 35 (78%) were of high-risk histology (G3, G4, or lymphatic invasion). Therefore, nm23-H1 expression within the primary tumor indicated the lymph node status with a sensitivity of 93% and a negative predictive value of 92%. The classic pathohistological factors (high risk vs low risk) had a sensitivity of 78% and a negative predictive value of 77%, respectively. CONCLUSION: Reduced expression of nm23-H1 within primary colorectal carcinomas could serve as an additional independent marker in estimating the nodal metastatic potential of these tumors. PMID- 9219796 TI - Thyroxine-binding globulin and thyroid hormones after resection of hepatocellular carcinoma. AB - OBJECTIVES: Some human hepatocellular carcinomas (HCCs) produce thyroxine-binding globulin (TBG). High serum TBG levels in such patients may be associated with increased thyroxine (T4) levels. This study aimed to elucidate the serum TBG and thyroid hormone profile in Japanese patients with HCC, to compare the difference between TBG-producing and -nonproducing HCCs, and to investigate the changes in serum TBG level and the thyroid hormone profile after removal of the tumor. METHODS: The 40 subjects included 20 patients with HCC, 10 healthy controls, and 10 operative controls. Serum TBG, 3,5,3'-triiodothyronine (T3), T4, and free T4 were measured serially for 4 wk after resection of HCC in 16 patients and after control operations in 10 patients. Assay methods were a radioimmunoassay for TBG and enzyme immunoassays for T3, T4, and free T4. RESULTS: Values higher than the mean +/- 2 SD of controls were considered abnormally high. Of patients with HCC, 60% had abnormally high TBG values, 65% had abnormally high T3 values, 39% had abnormally high T4 values, and 6% had abnormally high free T4 values. The mean levels of TBG and T3 were significantly higher than those in healthy controls, but no difference was found for T4 and free T4 levels. There were no significant differences in various clinicopathological factors between patients with high TBG levels and those with normal TBG levels. After resection of HCC, serum TBG decreased significantly in patients with high TBG levels but not in those with normal TBG levels. CONCLUSIONS: This study shows that >50% of HCCs in Japanese patients produce TBG; removal of the tumor reduces serum TBG in such cases. PMID- 9219797 TI - Cutaneous electrogastrography for the assessment of gastric myoelectrical activity in type I diabetes mellitus. AB - OBJECTIVE: Gastric dysrhythmias have been noted in diabetic patients with upper GI symptoms attributed to delayed gastric emptying. The aim of this study was to assess gastric myoelectrical activity in patients with asymptomatic insulin dependent diabetes mellitus. METHODS: Nine healthy subjects (five men, four women) and ten patients with insulin-dependent diabetes (six men, four women) participated in the study. Percutaneous electrogastrography was applied with a portable device on all subjects for 2 h before, during, and 2 h after the ingestion of a standard meal. Spectral analysis of the traces was performed on a personal computer using devoted software. The parameters assessed were 1) the percentile distribution of the three spectra of gastric slow-wave frequency, defined as follows: bradygastria for 0-2.4 cycles/min, normogastria for 2.5-3.6 cycles/min, and tachygastria for 3.7-9 cycles/min; and 2) the fed/fasting ratio of slow-wave power at all three spectra of frequencies. RESULTS: Bradygastria was significantly more common during the entire period of recording (p = 0.024), and in particular during the fasting state (p = 0.0008) and the period of meal consumption (p = 0.0001) in diabetic patients than in controls. In addition, the presence of normogastria did not increase significantly after the meal in diabetic patients as it did in controls. In diabetic patients, the slow-wave power decreased postprandially at the spectra of bradygastria and normogastria, unlike the controls, who exhibited a respective postprandial increase (fed/fasting power, controls vs patients:p = 0.0006 for bradygastria, p < 0.0001 for normogastria). CONCLUSIONS: Gastric dysrhythmias are present even in diabetic patients without upper GI symptoms attributed to gastric stasis. Increased presence of bradygastria and failure to increase the slow-wave amplitude postprandially are the predominant forms of abnormal myoelectrical activity in these cases. PMID- 9219798 TI - Dysphagia induced by chronic ingestion of benzodiazepine. AB - We report a case of a 57-yr-old woman with a complaint of dysphagia of 2-wk duration. She had rheumatoid arthritis and had been taking 10 mg of prednisone and 2 mg of lorazepam daily for 2 yr. Radiologic examination showed partial retention of barium sulfate in the pharynx, which was confirmed by scintigraphic examination of the oral and pharyngeal phases of swallowing. This retention was about 47% of the volume swallowed. The benzodiazepine was withdrawn, and 2 wk later she had no symptoms. We repeated the scintigraphic study, and it showed no retention. We conclude that chronic ingestion of benzodiazepine may cause dysphagia. PMID- 9219799 TI - Intrahepatic cholestasis due to systemic mastocytosis: a case report and review of literature. AB - A 35-yr-old female presented with symptoms of obstructive jaundice. Liver biopsy, bone marrow aspiration, and biopsy revealed systemic mastocytosis and acute myeloid leukemia. The liver biopsy specimen showed infiltration of mast cells within portal tracts with periductal and portal edema, irregularity of interlobular duct epithelium, and centrizonal cholestasis. Endoscopic retrograde cholangiography was normal. Following chemotherapy treatment with idarubicin and cytarabine for seven days for AML, the bilirubin levels continued to increase for two weeks and then decreased, reaching normal levels in two months. Infiltration of mast cells in the liver leads to hepatomegaly, liver function abnormality and rarely portal hypertension. Intrahepatic cholestasis due to systemic mastocytosis has never been reported. We report a rare case of systemic mastocytosis causing intrahepatic cholestasis that resolved with remission of AML following chemotherapy. PMID- 9219800 TI - Splenic rupture: an unusual complication of colonoscopy. AB - Splenic rupture is an uncommon complication of colonoscopy. A high index of suspicion is a crucial factor in the prompt diagnosis of this rare but potentially fatal complication. We report a case of splenic rupture diagnosed 3 days after a colonoscopy and requiring splenectomy. We also reviewed 17 reported cases of splenic rupture after colonoscopy, including our case. The presumed mechanisms of splenic rupture during colonoscopy are direct trauma to the spleen, excessive splenocolic ligament traction, and decrease in the relative mobility between the spleen and the colon. Of the 17 cases reviewed, 10 had polypectomy and/or biopsy performed during colonoscopy. Other probable risk factors are identified and tabulated. The hemodynamic status of the patient is the primary factor used to determine the therapeutic option. Computed tomographic (CT) scan of the abdomen reliably demonstrates well-contained splenic laceration and subcapsular hematoma, and differentiates these splenic complications from perisplenic clot and hemoperitoneum. Thus, CT scan may help decide which patients may be managed operatively or nonoperatively. Splenectomy is the operative procedure of choice for splenic rupture after colonoscopy. Conservative management includes broad spectrum antibiotics, intravenous fluids, blood transfusion, and close hemodynamic monitoring. The factors mandating further evaluation of persistent abdominal pain after colonoscopy are hemodynamic instability, clinical features of acute abdomen, leukocytosis, and/or acute anemia. The onset of abdominal pain associated with one or more of these critical factors is usually within 24 h after colonoscopy. An emergent CT scan of the abdomen is the modality of choice to further evaluate these clinical features, but intestinal perforation and external bleeding must first be excluded. PMID- 9219801 TI - Giant antral ulcer: a rare presentation of eosinophilic gastroenteritis--case report and review of the literature. AB - A 24-yr-old female presented with a giant gastric ulcer and anemia. She suffered from a transient infantile malabsorption syndrome with eosinophilia. The diagnosis of eosinophilic gastroenteritis associated with the gastric ulcer was made by endoscopic biopsy. Ulcer healing was refractory to medical therapy and partial gastrectomy was performed. Histologic examination revealed transmural eosinophilic infiltrates with mast cell infiltrates in the gastric wall. This case illustrates (1) an extremely rare presentation of eosinophilic gastroenteritis--giant, refractory, gastric ulcer; (2) a potential pathogenic role for mast cells in this syndrome; and (3) the chronic and relapsing nature of the syndrome. PMID- 9219802 TI - Gastroenteric inflammation in children with ulcerative colitis. AB - This is a retrospective review of five pediatric cases of inflammatory bowel disease with gastroduodenal as well as pancolonic inflammation. The presumptive diagnosis was Crohn's disease in all. Three of five also had microscopic ileitis. Chronic active gastritis was present at diagnosis in all five and duodenitis in four of five. None ever had noncaseating granulomas in any location. Prolonged (mean, 22 months) and ineffective trials of multiple medical therapies were carried out before subtotal proctocolectomy, which allowed the diagnosis of ulcerative colitis to be made in all children. Surgery remained curative after follow-up of over 1 yr (mean, 16 months). This experience confirms that gastroduodenal inflammation occurs in children with ulcerative colitis. Presence of gastroduodenal inflammation does not ensure the diagnosis of Crohn's unless other characteristic features of Crohn's are present. Accurate discrimination between Crohn's and ulcerative colitis remains important in management of pediatric inflammatory bowel disease to facilitate timely surgical referral. Upper gastrointestinal tract inflammation in ulcerative colitis warrants further study. PMID- 9219803 TI - Subtotal colectomy in a patient with collagenous colitis associated with colonic carcinoma and systemic lupus erythematosus. AB - A case of colonic carcinoma is described in a 60-yr-old woman with the typical clinical and histopathologic features of collagenous colitis and systemic lupus erythematosus, who is doing well 24 months after undergoing a subtotal colectomy with Brooke ileostomy. Collagenous colitis has only rarely been previously reported in association with colonic carcinoma (n = 1 case) or systemic lupus erythematosus (n = 1 case). In addition, this case is the second report of symptomatic collagenous colitis refractory to medical therapy, treated successfully with surgery. PMID- 9219804 TI - Extensive mesenteric inflammatory veno-occlusive disease of unknown etiology after primary cytomegalovirus infection: first case. AB - We report a young man who, shortly after a primary cytomegalovirus infection, presented with signs of intestinal ischemia requiring surgical intervention. The resected specimen of small bowel showed striking features of extensive phlebitis and venulitis affecting virtually all of the veins of the small intestine and mesentery. Although he had had a recent primary cytomegalovirus viremia, we could not identify any evidence of cytomegalovirus in the small bowel. He was not infected with HIV. The entity we describe is different from the recently reported mesenteric inflammatory veno-occlusive disease. The clinicopathologic entity represented by our patient's disease was heretofore unrecognized. PMID- 9219805 TI - Chromosome abnormality in solid and cystic tumor of the pancreas. AB - We report a case of a solid and cystic tumor of the pancreas with chromosomal abnormalities. The patient, a 13-yr-old girl, successfully underwent surgical excision and has been asymptomatic for more than 10 months. The tumor had two parameters suggesting malignant potential; a local invasion into a bile duct, and high mitotic activity. Chromosomal analysis showed two complex abnormalities, of which double loss of X chromosomes and trisomy for chromosome 3 were common. The complexity and polyclonality may stem from a clonal evolution. The observed abnormalities may provide a crucial clue to the understanding of the developmental process of this neoplasm. PMID- 9219806 TI - Use of octreotide in the treatment of mesenteric angina. AB - The case of a 57-yr-old man with a medical history of generalized atherosclerotic disease and newly diagnosed intestinal ischemia is presented. Because the patient was a poor surgical and anesthetic risk, medical treatment in the form of octreotide was administered with remarkable symptomatic relief. This is the first published report of the use of octreotide in treating mesenteric angina. PMID- 9219807 TI - Toxocara canis infection and granulomatous hepatitis. AB - This report describes possible involvement of Toxocara canis in granulomatous hepatitis in three patients who presented with varied clinical features; two were being treated for other diagnoses until Toxocara serology became available. Despite initial nonspecific therapy, there was clinical improvement in all patients and complete resolution of symptoms and abnormal liver biochemistry in two. T. canis infection should be considered in cases of granulomatous hepatitis. PMID- 9219808 TI - Chronic hepatitis related to use of fluoxetine. PMID- 9219809 TI - Transjugular embolization of the inferior mesenteric vein for bleeding anorectal varices after unsuccessful transjugular intrahepatic portosystemic shunt. PMID- 9219810 TI - Successful endoscopic injection sclerotherapy with N-butyl-2-cyanoacrylate following the recurrence of bleeding soon after endoscopic ligation for ruptured duodenal varices. AB - Bleeding from duodenal varices in a 63-yr-old man with alcoholic cirrhosis of the liver was found at endoscopy, and ligation surgery was carried out. Ten months after the operation, bleeding from the duodenal varices occurred and was treated by endoscopic ligation. However, after performing this procedure, bleeding again occurred 1 week later. Hemostasis was finally achieved by endoscopic injection sclerotherapy with N-butyl-2-cyanoacrylate. For the 22 months since the injection, the patient has been free from further bleeding. These results suggest that endoscopic injection sclerotherapy with N-butyl-2-cyanoacrylate was effective in bringing about immediate cessation of the bleeding in duodenal varices and that long-term hemostasis can be expected. PMID- 9219812 TI - Weeping umbilicus: an unusual presentation of abdominal tuberculosis. PMID- 9219811 TI - Plexiform neurofibromatosis and angiosarcoma of the liver in von Recklinghausen disease. PMID- 9219815 TI - Thrombocytopenia associated with hepatitis C: is it real? PMID- 9219814 TI - Double-contrast upper gastrointestinal examination with nonendoscopic biopsy: replacement for endoscopy? PMID- 9219813 TI - Reduction of abdominal hydatid disease with prolonged treatment. PMID- 9219816 TI - Equality of efficacy for open and laparoscopic antireflux repair? PMID- 9219817 TI - Mechanisms of acid clearance in reflux patients. PMID- 9219818 TI - Endoscopic retrograde cholangiography. PMID- 9219819 TI - Autoimmune hepatitis after cytomegalovirus infection in a bone marrow transplanted patient. PMID- 9219820 TI - Adult Kawasaki disease complicated by pancreatitis. PMID- 9219821 TI - Colorectal cancer risk in HNPCC families: development during lifetime and in successive generations. National Collaborative Group on HNPCC. AB - Members of hereditary non-polyposis colorectal cancer (HNPCC) families develop colorectal cancer at a much higher rate, and at a much younger age, than the general population. To quantify lifetime colorectal cancer risk in HNPCC family members, we calculated the cumulative incidence (CI) in different age categories, and compared this to the general population. Furthermore, we investigated whether successive generations of HNPCC families had earlier onset of disease. In 51 HNPCC families, selected according to the "Amsterdam criteria", the CI of colorectal cancer at age 75 was 40%, compared to only 4% in the general population. The CI ratio (CIR) of HNPCC family members relative to the general population was 148 at age 40, 79 at age 50 and 11 at age 75. Comparing successive generations of HNPCC families, the CI at age 75 increases from 19% in the ancestors to 32% in the first generation and 55% in the second generation. However, Cox proportional hazard analysis showed that this generation effect (RR per generation: 1.8, 95% CL = 1.4-2.2) largely disappears after adjustment for year of birth. In summary, at young ages, HNPCC family members experience an up to-150 times higher risk for colorectal cancer than the general population. This risk difference declines from age 60 onwards. The earlier age of onset in successive HNPCC generations does not appear to be a biological feature of HNPCC, but reflects a secular time trend in cancer occurrence in these families, similar to that in the general population. PMID- 9219822 TI - Evaluating cervical cancer screening programmes for developing countries. AB - This study evaluates cervical screening programmes for regions of the world where resources are scarce and little screening currently takes place. It investigates infrequent screening and programmes in which as many women as possible are screened just once in their lifetime. It also compares the effectiveness of cytology and human papillomavirus (HPV) testing for primary screening. Different programmes are evaluated by a stochastic model of the progression of pre-cancer, its relationship to papillomavirus infection and the diagnostic accuracy of alternative screening methods. These are compared in terms of the impact on the incidence of invasive cancer and resource use. Important factors that determine the suitability of different screening programmes are the available resources and the expected population coverage. Blanket screening for women aged 30-59 years, with the aim of covering all just once in their lifetime, could reduce the incidence of invasive cancer by up to 30%. A 10-year programme would require about 50% more routine screening tests to bring about the same reduction in incidence and a 5-year programme about 2.5-3 times as many. With either approach it would be more effective for resource use to concentrate on screening women aged 30-59 than a wider age group. Whether HPV testing would be more effective as a primary screening method than cytology depends on the underlying prevalence of HPV infection, the accuracy of cytology, the cost and the suitability of the testing procedure under field conditions. PMID- 9219823 TI - Distribution pattern of tenascin-C in normal and neoplastic mesenchymal tissues. AB - Descriptions for tenascin-C distribution are largely restricted to epithelial tumours. The present study utilized newly developed and characterized monoclonal (hT191) and polyclonal antibodies to investigate the distribution pattern of tenascin-C in a panel of mesenchymal tumours, which was contrasted with normal tissue. The specific antibodies recognized the distinctive star-like hexabrachion protein isolated from transformed cell-culture medium and serum from normal individuals. In normal tissues, a strong tenascin-C expression in the extracellular matrix was largely restricted to basement-membrane regions of epithelium and tonsilar sinusoids, pericellularly within smooth-muscle bundles, associated with perimysial, -chondrial, -neurial and -tendon surfaces, and diffusely within vascular adventitia. It was found in the corresponding tumours of the neural sheath (schwannoma) and smooth muscle (leiomyosarcoma), and was abundantly present around certain blood vessels of mesenchymal tumours. Although not detected in normal muscle, or in adipose or fibrous connective tissue, neo expression of tenascin-C was shown in more than half of the rhabdomyosarcomas, fibromas and liposarcomas, with an increased positive percentage in variably malignant myxoid liposarcomas compared with lipoma-like sarcomas. Tenascin-C was typically found in the extracellular matrix of soft-tissue tumours, but was notably absent from the epithelial-cell components of mixed epithelial/mesenchymal tumours. Its apparently enhanced expression in soft-tissue tumours differs from that of most other large extracellular-matrix proteins, suggesting possible functional involvement of the cell-adhesion molecule, tenascin-C, in the neoplastic phenotype. PMID- 9219824 TI - Prevalence of mutations and 30-bp deletion in the C-terminal region of Epstein Barr virus latent membrane protein-1 oncogene in reactive lymphoid tissue and non nasopharyngeal EBV-associated carcinomas in Hong Kong Chinese. AB - A specific variant of Epstein-Barr virus (EBV) with a 30-bp deletion in the C terminal region of the LMP1 gene has been found in some EBV-associated malignancies. To better understand the tumorigenic role of this LMP1 variant, we used PCR and sequencing to examine the LMP1 gene in 38 EBV-associated carcinomas (EBV-CAs) occurring in various organs (6 lung, 10 salivary gland, 5 sino-nasal, 16 gastric and 1 metastatic NPC), 55 reactive lymphoid tissues from tonsils (TON) and 67 EBV-negative tumours in various organs (22 adenolymphoma of salivary gland, 14 gastric and 31 colonic adenocarcinomas), where the virus was demonstrated in lymphocytes. The TON showed prevalence of both deleted and non deleted variants of LMP1, with dual infection being common. Significantly more of the LMP1 variant was deleted in EBV-CA and in EBV-negative tumours. Sequencing showed that the deleted and non-deleted variants have different sets of amino acid mutation. Mutations in codon 344 and 355 in the non-deleted variant disrupted the 9 nucleotide repeat flanking the deletion and thus may have conferred resistance to the deletion. The prevalence of both variants in the TON, with enrichment for the deleted variant in various organs, argues for the existence of an immune selection pressure in our population. The deleted variant, which may have a higher tumorigenic potential, may contribute to the high incidence of NPC, as well as the occurrence of EBV-CA in organs outside the nasopharynx in our locality. PMID- 9219825 TI - Comparison of the site distribution of melanoma in New Zealand and Canada. AB - A comparison of the site distribution of cutaneous malignant melanoma in New Zealand and Canada was performed. This series deals with 41,331 incident cases registered between 1968 and 1990 and is the largest to date to evaluate the influence of age and gender on the site distribution of melanoma. Site-specific, age-standardized rates per unit surface area and relative tumour density were assessed by gender and country and differences compared with statistical techniques adapted to this context. The age-standardized rates for all sites were higher in New Zealand than in Canada, the ratio being 3.2 for men and 3.8 for women. Occurrence of melanoma was denser for chronically than intermittently exposed sites in both New Zealand and Canada. The highest incidence rate per unit area was for the ears in men which was more than 5 times the rate for the entire body in each country. For each gender, melanomas were relatively commoner on the trunk and the face in Canada, and on the lower limbs in New Zealand. The variations in the site distribution were similar in each country and consistent with the effect of differential patterns of sun exposure between genders. Our results show that the levels of risk of melanoma between phenotypically comparable populations exposed to different amount of UV radiation vary in a site specific manner, especially for intermittently exposed sites. This suggests that both environmental conditions and lifestyle factors influence the site distribution of melanoma in these two populations. PMID- 9219826 TI - Human tenascin-C: identification of a novel type III repeat in oral cancer and of novel splice variants in normal, malignant and reactive oral mucosae. AB - Tenascin-C is a mosaic, linear glycoprotein that is up-regulated during many normal and pathological processes involving either cell migration or tissue morphogenesis, such as invasion of malignant cells and wound healing. Human tenascin-C contains 8 consecutive type III fibronectin (TNCfn) domains that are involved in alternative splicing and potentially generate a large number of isoforms that code for tenascin-C proteins with subtly different functions. Human tenascin-C splice variants were investigated by RT-PCR in a range of normal and pathological oral mucosal tissues. A novel, 9th human TNCfn domain involved in alternative splicing was identified. It shares 70% nucleic acid and 55% protein sequence homology with chicken TNCfn-ad2. As in avians, this novel repeat was located between TNCfn-B and TNCfn-ad1 and accordingly was designated human TNCfn ad2. Human TNCfn-ad2 was detected in only 2 of 10 oral cancers. However, TNCfn ad2 was absent from 40 normal, reactive, pre-malignant and other oral mucosal specimens investigated. Previous studies have described 8 splice variant transcripts for human tenascin-C. By systematic investigation we identified further novel splice variants for human tenascin-C. Furthermore, our results indicate that many potential splice variants probably do not exist in the tissues investigated. Thus, we have demonstrated that human tenascin-C transcripts generate a complex but selected repertoire of different alternative splice products. PMID- 9219827 TI - In situ T-cell responses in a primary regressive melanoma and subsequent metastases: a comparative analysis. AB - In an earlier study of the immune response in a patient with a cutaneous primary regressive melanoma, a T-cell-receptor diversity analysis demonstrated in situ amplification of certain lymphocytes. Two of them could be cloned and characterized as CD8+ HLA-class-l-restricted CTL with strong selective anti-tumor activity. Following a disease-free period of 3 years, the patient developed a gastric metastasis and subsequently (after an additional year) a metastasis in one axillary lymph node. Melanoma cell lines derived from the 2 secondary lesions have been established here. It was found that these metastatic cells have maintained expression of both HLA-class-I molecules and the peptidic antigen(s) recognized by the 2 clones amplified at the primary site. However, the corresponding T lymphocytes were either undetectable or poorly represented both in the gastric and in the axillary lesions. These results suggest that substantial alterations in the quality of T-cell infiltrates occurred during melanoma progression, despite an apparent stability in presentation of tumor associated antigen(s) which initially triggered a positive rejection response. PMID- 9219829 TI - Diet diversity and gastric cancer. AB - It has long been suggested that a varied diet may protect against gastric cancer, in the absence, however, of definition and quantification of the issue. Thus, we considered the relationship between diet diversity (i.e., variety of food intake computed as the total number of foods consumed at least once per week) and the risk of gastric cancer using data of a case-control study conducted between 1985 and 1993 in northern Italy on 746 gastric-cancer cases below age 75 years and 2,053 controls admitted to hospital for acute, non-neoplastic, non-digestive tract diseases. A significant inverse association was observed between various measures of food diversity and gastric cancer risk. Compared with subjects in the lowest quartile of total diversity, the multivariate odds ratios (ORs) were 0.9 for the second, 0.9 for the third and 0.7 for the highest quartiles. The inverse association was even stronger for vegetable (OR = 0.5 for the highest level) and fruit (OR = 0.6) diversity. Our findings were not explained by allowance for total calorie intake and total number of servings, besides education as an indicator of social class, and support, therefore, the concept that a more diversified and richer diet is a relevant underlying correlate of the decline in gastric cancer rates. PMID- 9219828 TI - Frequent rearrangements at minisatellite loci D1S7 (1p33-35), D7S22 (7q36-ter) and D12S11 (12q24.3-ter) in hepatitis B virus-positive hepatocellular carcinomas from Thai patients. AB - Primary hepatocellular carcinoma (HCC) is one of the most common cancers in Thailand; chronic infection with hepatitis B virus (HBV) is endemic and represents a major risk factor for the development of this cancer. Several mechanisms for HBV-related hepatocarcinogenesis have been proposed, among them a direct role of HBV in the promotion of genetic recombination leading to chromosomal alterations. Minisatellite DNA sequences are hypervariable regions dispersed throughout the genome which are susceptible to genetic recombination events. In the present study, somatic rearrangements affecting minisatellite sequences were examined in a total of 26 HCC from Thai patients. Multilocus DNA fingerprinting using probes 33.15 and 33.6 detected rearrangements in 11 and 12 HCC, respectively, all of them carrying integrated HBV DNA. The frequency of rearranged bands was calculated for each probe based on the total number of rearrangements observed in the 26 tumours and the total number of bands revealed by DNA fingerprinting in the non-tumour DNA. With each probe a total of 23 rearrangements was observed, yielding rearrangement frequencies of 3.7% and 4.2% for the 33.15 and 33.6 minisatellite families, respectively. To test for possible clustering of these rearrangements at specific loci, we used minisatellite locus specific probes previously cloned from 33.15 and 33.6. Minisatellites located at 1p33-35, 7q36-ter and 12q24.3-ter were shown to be frequently affected by rearrangement events in this series of HBV-positive HCC. Frequent rearrangements at minisatellite locus D7S22 (7q36-ter) in HBV-positive human HCC have not been reported so far. PMID- 9219830 TI - Low O-acetylation of sialyl-Le(x) contributes to its overexpression in colon carcinoma metastases. AB - Two factors potentially determining the consistent overexpression of sialyl-Le(x) antigen in colon carcinoma and metastases were investigated: (i) the expression of the mucins MUC1 and MUC2, known to carry sialyl-Le(x), by Northern blotting; (ii) the extent of sialic acid O-acetylation, by Western blotting and HPLC. RNA and sialyl-Le(x)-positive mucins were purified from normal colonic mucosa (N), primary carcinomas (T) and their liver metastases (M). Northern blots showed that mRNA expression both of MUC1 and of MUC2 decreases during the progression of the disease, and is lowest in metastatic tissue. The expression of mucin-bound sialyl Le(x) increased strongly from N to T and, to a lesser extent, to M. After alkali treatment of the mucins these differences disappeared, indicating that the total amount of mucin-bound sialyl-Le(x) is the same in the 3 types of tissues. The O acetylation of mucin-bound sialyl-Le(x) gradually decreased from N to M. HPLC analysis showed that in N about 70%, in T 45% and in M only 20% of mucin-bound sialic acids are O-acetylated. Thus, the increase of sialyl-Le(x) detectable during colon-carcinoma progression is due to diminished O-acetylation and not to increased expression of mucin protein cores. The decrease of O-acetylation is therefore the primary chemical alteration contributing to colon carcinoma associated overexpression of sialyl-Le(x). PMID- 9219831 TI - Genetics as a diagnostic tool in sarcomatoid renal-cell cancer. AB - Renal-cell cancer comprises a heterogeneous group of tumors, which currently can be sub-divided into morphologically distinct entities, each characterized by a specific combination of genetic changes. Sarcomatoid transformation might occur in any of the sub-types, resulting in tumors consisting of both carcinomatous and sarcomatous components. The specific diagnosis of these neoplasms, as to tumor sub-type, is usually made on the histologic properties of the carcinomatous tissue present. However, this might not reflect the true nature of the sarcomatous component. Since the genetic changes associated with the development of the different sub-types of renal-cell cancer are well established, this knowledge might serve as a tool in diagnosing sarcomatoid tumors. Assessing the genetic constitution of the latter may lead to correct diagnosis. It may also provide valuable information about the genetic changes associated with sarcomatoid transformation. Hence we performed a genetic characterization of a case of sarcomatoid renal-cell cancer, histologically diagnosed as being of the chromophilic type. The observed genetic changes included loss of 3p, 6q, 8p, 9, 13, 14 and 17p, and gain of 5, 12 and 20, as well as a mutation in the coding region of the p53 gene. This combination of genetic changes points to clear-cell rather than chromophilic origin of the sarcomatoid tumor investigated, indicating that the genetic constitution of sarcomatoid tumors may be a more reliable indicator of tumor sub-type than histologic appearance. PMID- 9219832 TI - Genomic deletion and p53 inactivation in cervical carcinoma. AB - The tumor-suppressor gene p53 acts as "the guardian of the genome", sensing DNA damage and initiating protective responses. To examine the hypothesis that p53 abnormality leads to increased genomic alterations in primary tumor cells, our study utilized 51 primary tumors of cervical carcinoma and 10 microsatellite markers. These markers were mapped to the short arms of chromosomes 3 and 5, covering the regions 3p13-25 and 5p15.1-15.3. Genomic deletion on 3p and 5p was correlated with genetic or epigenetic p53 inactivation pathways, including p53 mutation, genetic deletion of p53 and cervical infection with human papillomavirus. The proportion of abnormal p53 was found to be significantly higher in the cases exhibiting loss of heterozygosity (LOH) on 5p (p < 0.001), supporting the hypothesis of the presence of a p53-dependent pathway to cervical tumorigenesis. In contrast, however, LOH on 3p was found to be independent of p53 inactivation. A common deletion region, 3p22-24, was identified in 44% of informative cases, and genomic loss at this specific region was correlated with early tumorigenic onset and poor grade of tumor differentiation. Diversity within the patterns of genomic alteration in the same form of cancer suggests different sets of risk/tumorigenic profiles, molecular pathogenesis, as well as prognosis and outcome. PMID- 9219833 TI - Mutation analysis and loss of heterozygosity of PEDF in central nervous system primitive neuroectodermal tumors. AB - Deletion of 17p is the most frequent abnormality observed in central nervous system (CNS) primitive neuroectodermal tumors (PNETs), implicating the presence of a tumor suppressor gene which maps to 17p. The gene for pigment epithelium derived factor (PEDF) has been cloned and mapped to 17p13. PEDF belongs to the serine protease inhibitor (SERPIN) gene family. The PEDF protein has neurotrophic and neuronal-survival activities and is expressed in the CNS. Twenty tumor and matched normal DNA samples from patients with PNETs were screened by single strand conformation polymorphism (SSCP) analysis to determine loss of heterozygosity (LOH) and to identify potential mutations within the 8 exons of the PEDF gene. Ten of the 20 tumors demonstrated LOH, consistent with the deletion status of 17p determined by cytogenetic or fluorescence in situ hybridization studies. SSCP analysis of the genomic DNA from the 10 cases with LOH demonstrated several polymorphisms in exons 4 and 7, but no mutations. Our results are consistent with a loss of alleles on 17p in 50% of CNS PNETs, but do not suggest that PEDF is a candidate for the PNET suppressor gene in 17p13. PMID- 9219834 TI - Deletion of chromosome 11p15, p12, q22, q23-24 loci in human prostate cancer. AB - Loss of heterozygosity (LOH) on chromosome 11 is frequently altered in various epithelial cancers. The present study was designed to investigate LOH on chromosome 11 in microdissected samples of normal prostatic epithelium and invasive carcinoma from the same patients. For this purpose, DNA was extracted from the microdissected normal and tumor cells of 38 prostate cancers, amplified by polymerase chain reaction PCR and analyzed for LOH on chromosome 11 using 9 different polymorphic DNA markers (D11S1307, D11S989, D11S1313, D11S898, D11S940, D11S1818, D11S924, D11S1336 and D11S912). LOH on chromosome 11 was identified in 30 of 38 cases (78%) with at least one marker. Four distinct regions of loss detected were: 1) at 11p15, at loci between D11S1307 and D11S989; 2) at 11p12, on locus D11S131 (11p12); 3) at 11q22, on loci D11S898, D11S940 and D11S1818; and 4) at 11q23-24, on loci between D11S1336 and D11S912. We found 25% of the tumors with LOH at 11p15; 39% had LOH at 11p12; 66% had LOH at 11q22; and 47% had LOH at 11q23-24. These deletions at 11p15, 11p12, 11q22 and 11q23-24 loci were not related to the stage or grade of the tumor. PMID- 9219835 TI - Photodynamic activities and biodistribution of fluorinated zinc phthalocyanine derivatives in the murine EMT-6 tumour model. AB - The photodynamic properties and biodistribution pattern of zinc dodecafluoro-4 sulphophthalocyanine (ZnPcF12S1), zinc hexadecafluorophthalocyanine (ZnPcF16) and zinc phthalocyanine (ZnPc) were evaluated in the murine EMT-6 tumour model. All 3 dyes were formulated as a Cremophor oil-water emulsion after initial solubilization in methanol, acetone and pyridine, respectively. Comparison of their phototoxicity after in vitro incubation with EMT-6 cells and exposure to various fluences of red light showed that ZnPcF12S1 is about 50 times more active than ZnPcF16, reflecting better cell-penetrating properties. Solubilisation of ZnPc in 1-methyl-2-pyrrolidinone prior to formulation resulted in loss of photoactivity upon dilution in serum due to precipitation of the dye in the aqueous environment. In contrast, initial solubilisation in pyridine likely forms a ZnPc-pyridinium salt, and this preparation was 6 times more phototoxic than ZnPcF12S1. In vivo comparison of monosulphonated ZnPcF12S1 with perfluorinated ZnPcF16 showed improved pharmacokinetics in mice, including lower liver and spleen retentions and higher tumour-to-non-target tissue ratios. However, photodynamic therapy (PDT) of the EMT-6 tumour in BALB/c mice with red light, 24 or 48 hr post-injection of 1 micromol x kg(-1) of ZnPcF12S1 induced mortality. Lowering the drug and/or light dose or extending the time interval between drug administration and irradiation to 72 hr avoided adverse effects but also resulted in poor tumour response. The best tumour control (25% of animals) was obtained at 0.1 micromol x kg(-1) and a fluence of 400 J x cm(-2) at 24 hr post-injection. In contrast, ZnPcF16 required a 20-fold higher drug dose to induce a similar tumour response. The systemic shock following PDT with the amphiphilic ZnPcF12S1 likely results from extensive cellular effects. PMID- 9219836 TI - Overcoming CPT-11 resistance by using a biscoclaurine alkaloid, cepharanthine, to modulate plasma trans-membrane potential. AB - Irinotecan, 7-ethyl-10-[4-(1-piperidino)-1-piperidino] carbonyloxycamptothecin, (CPT-11) resistance was overcome by using a biscoclaurine alkaloid, cepharanthine, in CPT-11- and multidrug-resistant 50MT-1 cells. 50MT-1 cells were established from a mouse breast-cancer cell line, FM3A, by subjecting the cells to a low dose of CPT-11 continuously. 50MT-1 cells exhibited resistance to CPT-11 (40-fold in colony-formation assay) and to other drugs such as doxorubicin (11.7 fold) and etoposide (VP-16) (16.8-fold). The plasma trans-membrane potential was lower in 50MT-1 cells than in FM3A cells, although there were no differences in expressions of P-glycoprotein and of DNA topoisomerase-I and -II proteins. The lower membrane potential in 50MT-1 cells was augmented by co-treatment with a non toxic dose of cepharanthine. CPT-11 resistance in 50MT-1 cells was overcome (5.0- to 1.4-fold, 6-hr exposure) by the co-treatment with cepharanthine through increasing intracellular accumulation of CPT-11. Resistance to doxorubicin and VP 16 was also overcome by cepharanthine treatment (2.5- to 0.69-fold and 4.2- to 1.4-fold respectively). We conclude that the modification of plasma trans membrane potential by cepharanthine should be effective in overcoming CPT-11 and multidrug resistance in 50MT-1 cells. PMID- 9219837 TI - Response of the methionine synthase system to short-term culture with homocysteine and nitrous oxide and its relation to methionine dependence. AB - We compared the metabolic response of a methionine(Met)-dependent (P60) human glioma cell line with that of a Met-independent variant (P60H) when cultured in a homocysteine (Hcy) medium and exposed to N2O. In Hcy medium (without Met), remethylation of Hcy in P60H cells was enhanced and supported growth, whereas remethylation was low in P60 cells, which failed to thrive under these conditions. Both cell types seemed to contain adequate amounts of folates and total cobalamin (Cbl). P60 cells showed increased total and methylcobalamin (CH3Cbl) content after the shift to a Hcy medium, but the high, stable level of CH3Cbl detected in P60H cells was not attained. Further metabolic differences were induced by N2O exposure, which markedly reduced Met-synthase activity in cell-free extracts in both cell lines and completely blocked intact-cell Hcy remethylation in P60, whereas Hcy remethylation was only partly inhibited in P60H cells cultured in Met medium. The residual Hcy remethylation in P60H cells may be related to only a moderate depletion of CH3Cbl. The resulting high CH3Cbl level relative to Met-synthase activity during N2O exposure was even higher in Hcy medium. These findings in P60H cells probably reflect increased provision of Cbl to support Hcy remethylation under metabolic strain. The inability of P60 to furnish CH3Cbl to the enzyme may explain both the Met-dependent phenotype and the increased sensitivity of Hcy remethylation to N2O exposure in these cells. PMID- 9219838 TI - A comparison of p53 and p16 expression in human tumor cells treated with hyperthermia or ionizing radiation. AB - To assess the potential relationship between p53 and p16 proteins in the cellular response to stress, we have examined the levels of these proteins in a series of human tumor cell lines after treatment with either ionizing radiation or hyperthermia. We found that cells with abnormal radiation-induced G1 arrest (non functional p53) had significantly higher constitutive levels of p16 than cells showing a normal G1 arrest (functional p53). Time-course experiments were done to test the effect of gamma-irradiation on intracellular levels of p16. The pattern of changes in p16 response was similar in all cell lines studied, and p16 expression was not related to cellular sensitivity to radiation or to the level of p53 induction after treatment. We also provide evidence that short-term exposure to high temperature causes p53 accumulation. Hyperthermia-induced p53 accumulation was greatest in those cells exhibiting the highest radiation-induced p53 accumulation, suggesting a possible relationship between p53 induction after these 2 different stresses. p16 synthesis was also induced in different cell lines after heat treatment, and this response was independent of p53 functionality. When we compared the level of p16 expression with the extent of G0/G1 arrest induced by heat, a linear correlation was found, raising the possibility that p16 may be involved in the control of cell cycle progression in response to heat treatment. PMID- 9219839 TI - Establishment and characterization of 12 uterine cervical-carcinoma cell lines: common sequence variation in the E7 gene of HPV-16-positive cell lines. AB - A total of 12 carcinoma cell lines of the human uterine cervix were established from 5 keratinizing and 5 nonkeratinizing squamous-cell carcinomas, and 2 small cell carcinomas. Of these, 10 lines grew as adherent cells and 2 as floating aggregates. All lines showed (i) similarity in morphology to the primary tumor from which they were derived; (ii) high viability with relatively long doubling times (48-96 hr); (iii) absence of Mycoplasma and other bacteria, apart from one Mycoplasma-contaminated line; (iv) genetic heterogeneity by DNA-fingerprinting analysis; (v) absence of p53 mutation from exon 4 through 9; and (vi) the presence of HPV DNA sequence. Among the lines, 7 were infected by HPV-16, 3 by HPV-18, 1 by HPV-31, and 1 by HPV-33; the 2 cell lines derived from small-cell carcinomas contained HPV-18. Interestingly, 6 of the 7 cell lines containing HPV 16-type DNA harbored the same alteration of E7 at nucleotide position 647 (amino acid 29, AAT --> AGT, Asn --> Ser), whereas the 3 HPV-18-positive lines did not; 3 cell lines proved to have intact E1/E2 of HPV, suggesting the presence of episomally replicating HPV DNA as well as the integrated form, whereas the other 9 lines were shown to have integrated HPV. Taken together, these cell lines would be very useful for studying the biology of uterine cervical carcinoma. PMID- 9219840 TI - SV-IV, a major protein secreted from rat seminal vesicle epithelium, promotes lymphocyte cytotoxic activity against the lymphoblastoid Raji cell line in human peripheral blood mononuclear cells. AB - The treatment of human peripheral blood mononuclear cells (PBMC) with micromolar concentrations of SV-IV, a major protein secreted from the rat seminal vesicle epithelium, promotes in this cell population a marked cytotoxic activity against the Raji lymphoblastoid cell line. This activity is apparently due to cell-to cell contact interactions. The expression of HLA DR on Raji cells has a modulatory effect on the SV-IV-induced cytotoxic activity. The experimental evidence strongly suggests that the cytotoxic effector cells are functionally activated NK cells. PMID- 9219841 TI - Effect of chemical structure and hydrophobicity on the pharmacokinetic properties of porphycenes in tumour-bearing mice. AB - The efficiency and selectivity of tumour targeting by several tetra-n propylporphycene (TPPn) and tetrakis(methoxyethyl)porphycene (TMPn) derivatives have been studied by administering 3.76 micromol/kg of aqueous or liposomal porphycene formulations to BALB/c mice bearing an i.m. implanted MS-2 fibrosarcoma. These 2 parameters have been studied as a function of the type of substituents linked to the 9-position of the macrocycle by amide, ester or ether functional groups. The pharmacokinetic properties appear to be controlled mainly by the degree of porphycene hydrophobicity, as evaluated by measuring their retention times in a C 18 column for HPLC. Thus, the post-injection time (T50) at which the porphycene concentration in the plasma decreases to 50% of the initial value ranged from a few minutes for the less hydrophobic to several hours for the more hydrophobic porphycenes. An increase in hydrophobicity also was accompanied by an enhanced efficiency and selectivity of tumour targeting. The less hydrophobic porphycenes showed a maximum tumour uptake of 0.5-2 nmol/g of tissue at 10-20 min after administration with a tumour/peri-tumoural concentration ratio around 2-3, while those with higher hydrophobicity reached tumour concentrations of 7-8 nmol/g at 24-48 hr after administration with concentration ratios higher than 20. PMID- 9219842 TI - Lack of phospholipase A2 mutations in neuroblastoma, melanoma and colon-cancer cell lines. AB - A candidate murine tumor-suppressor gene, Mom1, has been identified as the secretory phospholipase A2 (GDB nomenclature: PLA2G2A) gene. Evidence suggests that PLA2G2A functions as a tumor-suppressor because mice lacking PLA2G2A expression demonstrate increased colonic polyposis. The human homologue of PLA2G2A has been mapped to chromosome 1p36, a region frequently implicated in the pathogenesis of neuroblastoma, colon cancer and melanoma. We identified 2 alterations in the PLA2G2A gene in a single neuroblastoma cell line out of 20 examined; however, we found no mutations in 24 melanoma cell lines, 12 lymphoblastoid cell lines from patients having chromosome 1-linked familial melanoma and 10 colon cancer cell lines. Secretory phospholipase A2 is unlikely to play a significant role in the pathogenesis of these tumors. PMID- 9219843 TI - Oxytocin inhibits the proliferation of MDA-MB231 human breast-cancer cells via cyclic adenosine monophosphate and protein kinase A. AB - Oxytocin (OT) inhibits the proliferation of breast-cancer cells in vitro via a specific G-coupled receptor. To elucidate the intracellular mechanism involved in this biological effect, different G-coupled receptor mediators have been investigated in untreated and OT-treated MDA-MB231 breast-carcinoma cells. In these cells, after OT treatment, a significant cAMP increase was observed using a radioimmunoassay procedure, whereas the Ca2+ (determined with the fluorescent probe fura-2) and the inositol phosphate (determined after cell labeling with myo(2-(3)H)-inositol) concentrations were not modified, contrary to what has been observed in myometrial and myo-epithelial cells. The PKA inhibitor PKI (6-22) amide reverted the effect of OT, indicating that the anti-proliferative effect of the peptide is strictly related to the cAMP-PKA pathway. OT treatment did not modify tyrosine phosphorylation either. Our results indicate that in breast epithelial cells devoid of contractile activity, cAMP is the intracellular mediator of OT action, whereas the Ca2+-phosphoinositide system is not involved. PMID- 9219844 TI - Characterization of a cAMP-dependent protein kinase mutant resistant to cisplatin. AB - The signal transduction pathway of cAMP, mediated by the cAMP-dependent protein kinase (PKA), is involved in the regulation of metabolisms, cell growth and differentiation and gene expression. Isolated PKA mutants from Chinese hamster ovary (CHO) cells were used in our laboratory to study the role of cAMP in the development of drug resistance in cancer. We have found that PKA mutants harboring a defective regulatory (RI alpha) subunit, but not the catalytic (C) subunit, are more resistant to the chemotherapeutic drug cisplatin. To clarify the role of PKA in cisplatin resistance, we have performed a step-wise selection with a CHO RI alpha subunit mutant cell line, 10248, for further resistance to cisplatin. A representative clone (10248/CDDP(R)-5) was used for further characterization. These cisplatin-resistant PKA mutant cells remained refractory to cAMP-induced growth inhibition and had decreased PKA activity comparable to the parental 10248 mutant cells. Furthermore, 10248/CDDP(R)-5 also exhibited cross-resistance to the nitrogen mustard melphalan but maintained the same sensitivity as wild-type cells to non-DNA-damaging agents such as methotrexate. The mechanism of resistance may be due to increased DNA repair as assessed by the host cell reactivation assay. We speculate that mutation and functional inactivation of PKA may result in deregulated growth response to cAMP, as well as the acquisition of resistance to cisplatin and other DNA-damaging agents in cancer. PMID- 9219845 TI - Protection of 5alpha-dihydrotestosterone against TGF-beta-induced apoptosis in FaO cells and induction of mitosis in HepG2 cells. AB - Administration of TGF-beta1 to both FaO and HepG2 cells significantly induced apoptosis, particularly in FaO cells. Degradation of genomic DNA in FaO cells was rapidly induced by treatment with TGF-beta1 (5 ng/ml) for only 4 hr. 5alpha dihydrotestosterone (DHT, 25 nM) alone did not affect any significant changes in cell viability and in nuclei of FaO cells; however, pre-treatment with DHT protected genomic DNA degradation induced by TGF-beta1 for 14 hr. Simultaneous treatment with DHT plus TGF-beta1 (D + T) inhibited TGF-beta-induced apoptosis by approximately 50% in FaO cells. On the other hand, D + T treatment increased mitosis in actively growing HepG2 cells. Thus, it is reasonable to conclude that DHT gives growth advantage to hepatocellular-carcinoma cells by inhibiting TGF beta-induced DNA fragmentation in FaO cells and by inducting mitosis in HepG2 cells. PMID- 9219846 TI - Transforming growth factor-beta1 enhances the lethal effects of DNA-damaging agents in a human lung-cancer cell line. AB - In tissue culture conditions, exogeneous active transforming growth factor-beta1 (TGF-beta1) enhances the lethal effect of DNA-damaging agents (UV-C, gamma rays, cisplatin, methotrexate and 5-fluorouracil) toward human A549 cells and mink Mv1Lu cells, as detected by the loss of their capacity to give rise to colonies; both these cell lines harbor a wild-type p53, as determined by immunoprecipitation. Contrastingly, the sole effect of the cytokine used alone is to inhibit reversibly the multiplication of the same cells without further impairing, once withdrawn from their environment, their capacity to divide and give rise to colonies. The lethal synergy between TGF-beta1 and UV-C was studied on mink and human cell lines, and the biomodulation by TGF-beta1 of cell killing by cisplatin, gamma rays, 5-fluorouracil or methotrexate was tested only on human cells. As investigated with UV-C-irradiated human A549 cells, TGF-beta1 appears to enhance apoptosis rather than to disturb the repair of DNA photolesions (mainly pyrimidine dimers) by the nucleotidic excision repair pathway according to results of nucleosomal ladder and comet tests. Our data raise the possibility that, in vivo, TGF-beta1 might affect the curative and/or undesirable secondary side effects of cancer therapy. PMID- 9219847 TI - Regulation of cell spreading during differentiation in the muscarinic M5 receptor tumor-suppressor model. AB - Activation of the muscarinic receptor in Chinese hamster ovary (CHO) cells results in a reversal of the malignant phenotype for which spreading into a bipolar, fibroblastic morphology is a marker. The process of morphologic change requires multiple events, including alterations in adhesions to substrates and cytoskeletal re-arrangement. In this report, we demonstrate the calcium-dependent involvement of p125FAK in this cellular shape change using an inhibitor of ligand induced calcium influx, carboxyamido-triazole (CAI). p125FAK becomes tyrosine phosphorylated after exposure to the agonist carbachol (CC), reaching maximal phosphorylation prior to initiation of cellular shape change at 1 hr into CC exposure (386 +/- 103%). Phosphorylation remained elevated through the shape change (4-12 hr). CHOm5 cell exposure to the Ca2+-mobilizing agents maitotoxin and ionomycin also resulted in p125FAK phosphorylation. Inhibition of Ca2+ influx with CAI, an inhibitor of ligand-induced Ca2+ influx, had little effect on CC induced phosphorylation but partially inhibited ionomycin-mediated p125FAK phosphorylation. While the intracellular Ca2+ chelator BAPTA failed to prevent CC induced p125FAK tyrosine phosphorylation, it inhibited phosphorylation due to ionomycin. CC induced Ca2+-independent binding of phosphorylated p125FAK selectively to the C-terminal SH2 domain of phosphatidylinositol-3'-kinase (PI3K). Further, CC, maitotoxin and ionomycin induced in vitro kinase activity of p125FAK for the exogenous substrate poly(Glu4Tyr1). Kinase activity stimulated by all 3 agonists was inhibited by preincubation with either CAI or BAPTA. Our results indicate that increasing intracellular Ca2+ can stimulate both p125FAK autophosphorylation and kinase activity; however, p125FAK phosphorylation in response to CC also may be induced through a Ca2+-independent pathway. PMID- 9219848 TI - Stromal fibroblasts influence oral squamous-cell carcinoma cell interactions with tenascin-C. AB - In this study we identified tenascin-C (TN-C) and one of its integrin receptors, alpha(v)beta6, in oral squamous-cell carcinoma (SCC) specimens. Neither TN-C nor alpha(v)beta6 are expressed in normal oral mucosa. We also studied 2 human oral squamous-cell carcinoma cell lines: the highly invasive HSC-3 cells, and the poorly invasive SCC-25 cells. We determined that adhesion of these cells to TN-C involves both alpha2 and alpha(v) integrins. Migration on TN-C by oral SCC cells required fibroblast-conditioned medium and did not occur in its absence. This migration was blocked by anti-alpha2 and anti-alpha(v) antibodies and was partially inhibited by antibodies to hepatocyte growth factor, epidermal growth factor and transforming growth factor-beta1. When seeded on TN-C, the poorly invasive SCC-25 cells formed alpha(v)beta6-positive focal contacts; the HSC-3 cells did not. HSC-3, SCC-25 and PTF cells secrete TN-C into the culture medium, as determined by Western blot. However, when HSC-3 cells were inoculated into the floor of the mouth of nude mice, only murine TN-C could be identified in the reactive stroma adjacent to the resulting tumor nests, demonstrating that in vivo, HSC-3 cells do not secrete TN-C. Our results demonstrate that alpha(v)beta6 and tenascin-C are neo-expressed in oral squamous-cell carcinoma, and that the tumor stromal environment is influential in oral SCC behavior. PMID- 9219849 TI - Decreased sensitivity of carcinoembryonic antigen cDNA-transfected cells to adriamycin. AB - Carcinoembryonic antigen (CEA) is a heavily glycosylated protein and is expressed at a high frequency in adenocarcinomas, which are known to be one of the cancers most resistant to chemotherapeutic agents. In this study, with the aim to elucidate whether CEA participates in drug resistance or not, we tested the adriamycin (ADR) sensitivity of CEA transfectants of H-ras-transformed NIH 3T3 cells in vitro and in vivo. The ADR sensitivity of CEA transfectants in vitro was evaluated as growth (% of control) when incubated with various concentrations of ADR, and showed that they were higher than those of mock transfectants. The decreased ADR sensitivity of CEA transfectants in vivo was also observed as an increase in tumor size after intraperitoneal administration of ADR into SCID mice. To define the mechanisms for resistance, the accumulation and efflux of ADR in transfectants was examined. The rate of ADR accumulation in CEA transfectants was reduced compared to mock transfectants, caused at least partly by an increased efflux out of the cells. Furthermore, the modification of N-glycan on CEA by deoxymannojirimycin, an N-glycosylation processing inhibitor, partially restored ADR sensitivity of CEA transfectants, suggesting an involvement of sugar chains. Our data suggest that CEA expression may decrease ADR sensitivity of cancer cells. PMID- 9219850 TI - Age of menarche, birthweight and risk of cancer. PMID- 9219851 TI - An in vitro model of traumatic neuronal injury: loading rate-dependent changes in acute cytosolic calcium and lactate dehydrogenase release. AB - We developed a new in vitro model of neuronal injury using NT2-N cells to examine the effects of hydrodynamic loading rate on intraneuronal calcium dynamics and lactate dehydrogenase (LDH) release. Our apparatus consisted of a parallel disk viscometer which induced fluid shear stress with well-defined magnitudes and loading rates to cultured cells. We found that the deformation response of the cells was dependent on the severity of the insult, with increased cellular strains generated for higher shear stresses at a constant loading rate. Peak intracellular free calcium concentration correlated with strain, suggesting that mechanical deformation may regulate calcium response. Slowly applied fluid shear stress elicited no response, whereas high loading rates resulted in peak calcium increases 2.9 to 3.6 times baseline values as injury severity was increased. LDH release measured within 5 min after the insult correlated with loading rate. In addition, LDH release continued to increase out to 24 h following high loading rate conditions, demonstrating that the application of fluid shear stress led to prolonged cell damage. The acute response in NT2-N cells subjected to an insult with the CSID is dependent on the loading rate, and these results suggest that initial membrane deformation may trigger subsequent events. PMID- 9219852 TI - Immunohistochemical study of calpain-mediated breakdown products to alpha spectrin following controlled cortical impact injury in the rat. AB - This study examined the effect of unilateral controlled cortical impact on the appearance of calpain-mediated alpha-spectrin breakdown products (BDPs) in the rat cortex and hippocampus at various times following injury. Coronal sections were taken from animals at 15 min, 1 h, 3 h, 6 h, and 24 h after injury and immunolabeled with an antibody that recognizes calpain-mediated BDPs to alpha spectrin (Roberts-Lewis et al., 1994). Sections from a separate group of rats were also taken at the same times and stained with hematoxylin and eosin. Analyses of early time points (15 min, 1 h, 3 h, and 6 h following injury) revealed alpha-spectrin BDPs in structurally intact neuronal soma and dendrites in cortex ipsilateral to site of injury that was not present in tissue from sham injured control rats. By 24 h after injury labeling was not restricted to clearly defined neuronal structures in ipsilateral cortex, although there was an increased extent of diffuse labeling. BDPs to alpha-spectrin in axons were not detected until 24 h after injury, in contrast to the more rapid accumulation of BDPs observed in neuronal soma and dendrites. The presence of BDPs to alpha spectrin in the cortex at the site of impact, and in the rostral and contralateral cortex, coincided with morphopathology detected by hematoxylin and eosin. alpha-Spectrin BDPs were also observed in the hippocampus ipsilateral to the injury in the absence of overt cell death. This investigation provides further evidence that calpain is activated after controlled cortical impact and could contribute to necrosis at the site of injury. The appearance of calpain mediated BDPs at sites distal to the contusion site and in the hippocampus also suggests that calpain activation may precede and/or occur in the absence of extensive morphopathological changes. PMID- 9219853 TI - Neuronal cell loss in the CA3 subfield of the hippocampus following cortical contusion utilizing the optical disector method for cell counting. AB - Unilateral cortical contusion in the rat results in cell loss in both the cortex and hippocampus. Pharmacological intervention with growth factors or excitatory neurotransmitter antagonists may reduce cell loss and improve neurological outcome. The window of opportunity for such intervention remains unclear because a detailed temporal analysis of neuronal loss has not been performed in the rodent cortical contusion model. To elucidate the time course of hippocampal CA3 neuronal death ensuing cortical contusion, we employed the optical disector method for assessing the total number of CA3 neurons at 1 and 6 hours, 1, 2, 10, and 30 days following injury. This stereological technique allows reporting of total cell numbers within a given region and is unaffected by change in the volume of the structure or cell size. A rapid and significant reduction in neurons/mm3 in the ipsilateral CA3 field was observed by 1 h following trauma. However, a significant increase in neurons/mm3 was seen at 30 days postinjury. This surprising finding is a result of CA3 volume shrinkage and redistribution of CA3 neurons. Utilization of the optical disector reveals that regardless of an increase in neurons/mm3 at 30 days following injury, CA3 cell loss reaches 41% of control animals by 1 day posttrauma and remains near that level at all subsequent time points examined. It is estimated that there are about 156,000 neurons in the CA3 region in control animals. By 1 h following cortical contusion the cell population decreases to 93,000 neurons indicating a very rapid cell loss. This suggests a window of less than 24 h for pharmacological intervention in order to save CA3 neurons following cortical contusion. PMID- 9219854 TI - Whole body metabolic responses to brain trauma in the rat. AB - Increases in metabolic rate reported in head-injured patients can contribute to increases in respiratory demand, raised body temperature, and host body wasting (cachexia). The objective of the present study was to quantify the metabolic responses to brain trauma in the rat and investigate the underlying mechanisms. Lateral fluid-percussion (FP) injury (applied cortical pressure 1.6-1.8 atm) in the rat resulted in consistent and reproducible cortical brain lesions (44 +/- 6 mm3). Body weight and food intake were reduced significantly 24 h after brain trauma compared to sham-operated (7 and 49%,p < 0.01) and control animals (14 and 65%,p < 0.001), respectively. Resting oxygen consumption (V(O2), measured at 24 degrees C) was increased significantly, by 9-16% above sham-operated, and 14-26% above control animals for 2-7 h after brain trauma (p < 0.05), but V(O2) was not raised thereafter (24-72 h) and colonic temperature was not changed. Raising the ambient temperature from 24 degrees C to 28 degrees C significantly reduced the hypermetabolism of brain-injured rats compared to sham-operated controls. Injection of the beta-adrenoceptor antagonist propranolol (10 mg/kg, i.p.) completely abolished the rise in metabolic rate of brain-injured rats, and reduced significantly the rise in metabolic rate of the sham-operated animals (26%, p < 0.01 and 11%, p < 0.05; respectively). Systemic injection of the cyclo oxygenase inhibitor indomethacin (1 mg/kg, i.p.) significantly attenuated (by 11%,p < 0.01), but did not completely abolish the hypermetabolism of brain injured animals. Lateral FP injury in the rat causes a significant cachexia. Weight loss is due to hypophagia, and an increase in energy expenditure, which is mediated by sympathetic activation of thermogenesis and in part by prostaglandins. PMID- 9219855 TI - The neuroprotective effect of the forebrain-selective NMDA antagonist CP101,606 upon focal ischemic brain damage caused by acute subdural hematoma in the rat. AB - The neuroprotective effects of drugs that act against excitotoxic damage, caused by glutamate, are well described in focal ischemia, but behavioral effects, and apparent failure in clinical trials of "first-generation" competitive N-methyl D aspartate (NMDA) antagonists, such as Selfotel (CGS19755), has led to interest in evaluating newer NMDA antagonists with fewer behavioral effects. We have therefore evaluated the neuroprotective effect of a new forebrain-selective polyamine site NMDA antagonist, CP101,606 in a rat subdural hematoma (SDH) model. An SDH was produced by slow injection of 0.4 ml autologous blood into the parietal subdural space. Brain damage was assessed histologically at eight coronal planes, in animals sacrificed 4 h after induction of hematoma. The drug was infused 30 min after induction of SDH. The reductions of ischemic brain damage achieved by CP101,606, was 29% for the low dose and 37% for the high dose. This novel glutamate antagonist has shown a magnitude of neuroprotection which is comparable with that seen with "first-generation" NMDA antagonists such as MK801, D-CPP-ene and CGS19755, in this same model. This new agent is claimed to have fewer psychomotor and behavioral effects than MK801, D-CPP-ene, and CGS19755. PMID- 9219857 TI - Systemic administration of MPTP induces thalamic neuronal degeneration in mice. AB - 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a known neurotoxicant primarily selective for catecholaminergic neurons, including those of the nigrostriatal dopaminergic system, thereby mimicking the pathology of Parkinson's disease (PD). In this study, serial transbrain sectioning, followed by staining with a newly developed fluorochrome (Fluoro-Jade) specific for degenerating neurons, was used to detect additional sites of MPTP-induced neuronal degeneration in mice. Male CD-1 mice received a single 50 mg/kg dose of MPTP intraperitoneally at room temperature or at a reduced temperature (6 degrees C), which has been shown to potentiate striatal dopamine depletion. Neuronal degeneration was observed in the substantia nigra pars compacta (SN), ventral tegmental area (VTA) and retrorubral field (RRF) of only animals dosed in the low temperature environment. Neuronal degeneration was also observed in other catecholaminergic nuclei in both treatment groups. In addition, degenerating cell bodies and fibers were detected in the midline and intralaminar thalamic nuclei of all dosed animals, regardless of the dosing environment. Pharmacological manipulations which prevented nigral degeneration (deprenyl and nomifensine pretreatment) also prevented the degeneration of thalamic neurons. MK-801 pretreatment, however, resulted in a disproportionate protection of the thalamic neurons. These findings confirm and extend our previous observations regarding the protective effect of hyperthermia in CD-1 mice and also suggest that regions of the thalamus may be relevant to the pathophysiology of PD. PMID- 9219856 TI - Neuroprotective effects of the strychnine-insensitive glycine site NMDA antagonist (R)-HA-966 in an experimental model of Parkinson's disease. AB - The neuroprotective effects of (R)-HA-966 and (S)-HA-966 (3-amino-1-hydroxy-2 pyrrolidinone) were examined in an MPTP (1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine)-induced animal model of Parkinson's disease. Systemic pretreatment of C57 black mice with the strychnine-insensitive glycine site antagonist, (R)-HA-966 (3-30 mg/kg, i.p.), dose-dependently attenuated MPTP induced depletion of striatal dopamine and 3,4-dihydroxyphenylacetic acid (DOPAC). Pretreatment with (R)-HA-966 also significantly protected the degeneration of tyrosine hydroxylase-positive neurons in the substantia nigra of mice treated with MPTP and alleviated the acute behavioral changes caused by the neurotoxin. In contrast, the other racemic form, (S)-HA-966, neither prevented the neurochemical depletions nor the neuronal injury caused by MPTP. These results indicate that excitatory mechanisms of neurodegeneration are involved in the pathophysiology of Parkinson's disease, and that strychnine-insensitive glycine site NMDA antagonists may serve as dopaminoprotective agents which intervene in the progressive neurodegeneration in Parkinson's disease. PMID- 9219858 TI - Proconvulsive effect of vasopressin; mediation by a putative V2 receptor subtype in the central nervous system. AB - Subcutaneously (s.c.) administered [Arg8]vasopressin (AVP) potentiated seizures induced by intracerebroventricular (i.c.v.) injection of 1.95 mg pilocarpine (a muscarinic cholinergic agonist). A bell-shaped relation between dose and effect was found. I.c.v. pretreatment with a V1, V2 or oxytocin receptor antagonist was performed to determine whether and what type of receptor is involved in this proconvulsive effect of vasopressin. For these experiments a higher dose of pilocarpine (2.4 mg i.c.v.) was injected. This caused seizures in a slightly but not significantly higher percentage of the rats. A dose-dependent protective action of the V2 receptor antagonist d(CH2),[D-Ile2,Ile4]AVP (effective doses were 25 and 125 ng) on seizures was found. A reduction was observed in the number of animals that developed tonic-clonic convulsions. Neither the V1 receptor antagonist d(CH2)5[Tyr(Me)2]AVP nor the oxytocin receptor antagonist desGly(NH2)9d(CH2)5[Tyr(Me)2Thr4]OVT possessed anti-convulsive activity. Subsequently the type of receptor was studied in detail with fragments of AVP with either V1 or V2 activity. AVP (with V1 and V2 affinity) (1 and 3 microg s.c.) potentiated pilocarpine (1.95 mg) induced seizures. Vasotocin and oxytocin were without effect. Interestingly neither s.c. nor i.c.v. administration of the selective kidney type vasopressin receptor (V2) agonist dDAVP potentiated pilocarpine induced seizures. Several selective antidiuretic agonists (V2), such as d[Val4]AVP, d[Phe2,Val4,D-Arg8]vasopressin (3 microg), [Val4,D Arg8]vasopressin (3 microg) and d[Val4,D-Arg8]vasopressin (3 microg) were active. Other selective antidiuretic compounds, such as [Val4]AVP, dAVP, d[Tyr(Me)2]AVP and HO[D-Arg8]vasopressin (3 microg) did not influence seizures. These results demonstrate that a combination of substitution of aminoacid 4 (Gln) by Val and to a lesser extent deamination and the D-arginine form yield an active molecule, which can potentiate pilocarpine induced seizures and suggest the existence of a V2 receptor subtype in the brain. PMID- 9219859 TI - In vivo intracellular characteristics of inferior colliculus neurons in guinea pigs. AB - Intracellular in vivo recordings of physiologically identified inferior colliculus central nucleus (ICc) auditory neurons (n = 71) were carried out in anesthetized guinea pigs. The neuronal membrane characteristics are described showing mainly quantitative differences with a previous report [Nelson, P.G. and Erulkar, S.D., J. Neurophysiol., 26 (1963) 908-923]. The spontaneous spike activity was consistent with the discharge pattern of most extracellularly recorded units. The action potentials showed different spike durations, short and long, and some of them exhibited hyperpolarizing post-potentials. There were also differences in firing rate. The ICc neurons exhibited irregular activity producing spike trains as well as long silent periods (without spikes). Intracellular current injection revealed membrane potential adaptation and shifts that outlasted the electrical stimuli by 20-30 ms. Both evoked synaptic potentials and the spike activity in response to click and tone-burst stimulation were analyzed. Depolarizing-hyperpolarizing synaptic potentials were found in response to contralateral and binaural sound stimulation that far outlasted the stimulus (up to 90 ms). When ipsilaterally stimulated, inhibitory responses and no-responses were also recorded. Although few cells were studied, a similar phenomenon was observed using tone-burst stimulation; moreover, a good correlation was obtained between membrane potential shifts and the triggered spikes (input-output relationship). These in vivo results demonstrate the synaptic activity underlying many of the extracellularly recorded discharge patterns. The data are consistent with the known multi-synaptic ascending pathway by which signals arrive at the ICc as well as the descending corticofugal input that may contribute to the generation of long duration post-synaptic potentials. PMID- 9219860 TI - The role of nitric oxide in the pathophysiology of thromboembolic stroke in the rat. AB - Although nitric oxide (NO) has been shown to play an important role in the pathophysiology of cerebral ischemia, its contribution to the pathogenesis of experimentally induced thromboembolic stroke is unknown. In this study, we pharmacologically manipulated NO levels in the acute post-thrombotic stage and determined the effects on behavior and histopathology. The following drugs were used: nitro-L-arginine-methyl ester (L-NAME), a non-specific endothelial and neuronal nitric oxide synthase (eNOS and nNOS) inhibitor, 3-bromo-7-nitroindazole (7-NI), a specific inhibitor for nNOS, the NO precursor, exogenous L-arginine and the NO-donor, 3-morpholino-sydnonimine (SIN-1). Male Wistar rats (n = 76) were randomly assigned to receive vehicle or drug immediately after common carotid artery thrombosis (CCAT). Regional measurements of cortical NOS activity using the [3H]L-arginine to [3H]L-citrulline conversion assay were decreased 1 h after treatment with L-NAME and 7-NI by 50 and 65%, respectively; hippocampal NOS activity was reduced with L-NAME by 35% and with 7-NI by 65%. L-NAME significantly worsened forelimb placing as compared to other groups. 7-NI accelerated sensorimotor recovery. Water maze retention deficits were noted 48 h after CCAT and these were exacerbated by L-NAME treatment. Histopathological protection was conferred in the hippocampus by 7-NI and SIN-1; conversely, L-NAME increased neuronal injury in the contralateral cortex. L-arginine had no effect on these outcomes. In conclusion, both structural and functional consequences of CCAT can be aggravated by limiting endothelial NO production in the acutely post thrombotic brain. In contrast, inhibition of nNOS and infusion of an NO donor has a beneficial effect on pathology. PMID- 9219861 TI - Adenosine A2A receptor agonists increase Fos-like immunoreactivity in mesolimbic areas. AB - Expression of the early-gene c-fos is an useful method for studying potential sites of action of drugs active in the CNS. Stimulation of adenosine A2A receptors by CGS 21680 (5 mg/kg) induced an increase in Fos-like immunoreactivity in the rat nucleus accumbens shell, while in the rostral pole and core CGS 21680 induced Fos-like immunoreactivity only after a high dose. CGS 21680 (5 mg/kg) stimulated c-fos expression also in the lateral septal nucleus and dorso-medial striatum, but not in the dorso-lateral striatum. A similar pattern of Fos-like immunoreactivity was obtained after administration of the A2A agonist HENECA (5 mg/kg) which displays higher selectivity for A2A receptors than CGS 21680. Administration of the selective A2A antagonist SCH 58261 counteracted CGS 21680 induced Fos-like immunoreactivity. Lesions of the dopaminergic mesostriatal projection by 6-hydroxydopamine and stimulation of dopamine D2/D3 receptors by quinpirole, prevented CGS 21680-induced Fos-like immunoreactivity in the nucleus accumbens shell. The present results show that stimulation of A2A receptors induces a profile of c-fos expression similar to that of atypical neuroleptics. A2A receptor stimulation has been reported to have dopamine antagonistic actions, it is therefore suggested that A2A agonists might have antipsychotic activity without producing extrapyramidal side effects. PMID- 9219862 TI - Cytokine and growth factor immunohistochemical spinal profiles in two animal models of mononeuropathy. AB - Nerve injury leads to central neuroimmunologic responses that may be integral to the development and maintenance of chronic neuropathic pain in humans. Recent data have demonstrated that cytokines and growth factors may be strongly implicated in the generation of pain states at both peripheral and central nervous system sites. We utilized immunohistochemical methods to investigate this phenomenon in rat models of neuropathic pain. Specifically, we employed well characterized models of neuropathy that result in behaviors suggestive of neuropathic pain in humans; a freeze lesion of the sciatic nerve, termed sciatic cryoneurolysis, and a chronic constriction sciatic nerve injury. We used immunohistochemistry to examine spinal localization of the cytokines, interleukin 1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha) and the growth factors, basic fibroblast growth factor (bFGF), and transforming growth factor-beta1 (TGF beta) at 3, 14, and 35 days following sciatic cryoneurolysis or 6 days following chronic constriction injury as compared with normal, unoperated rats. There was minimal, diffuse cytokine/growth factor staining in lumbar spinal tissue from the normal group. However, cell profile quantification demonstrated increases in lumbar spinal IL-1beta-, TNF-alpha- and TGF-beta-like immunoreactivity (LI) in both mononeuropathy models studied. At 3 days following sciatic cryoneurolysis, intense bFGF LI was present in the ipsilateral dorsal and ventral horn. By 14 days bFGF LI was also observed in contralateral dorsal and ventral horns. In contrast, we found no obvious staining differences in lumbar spinal cord following the chronic constriction injury. This study demonstrated increased specific cytokine and growth factor-like expression in the spinal cord following peripheral nerve injuries. It also showed a differential expression of bFGF in two distinct mononeuropathy models. These results provide further evidence that central cytokine production via a neuroimmune cascade may be involved in the development and maintenance of behaviors that mimic neuropathic pain following nerve injury. PMID- 9219863 TI - Peripheral nerve regeneration through silicone chambers in streptozocin-induced diabetic rats. AB - The silicone chamber model was used to evaluate peripheral nerve regeneration (PNR) in streptozocin (STZ)-induced diabetic rats. Diabetic and control animals underwent sciatic nerve transection and silicone chamber implantation establishing gaps of various lengths between the transected nerve ends. In animals with 5 and 10 mm gaps, diabetes was induced in experimental rats 1 week before surgery, and the animals were sacrificed 3 weeks after surgery. In animals with 8 mm gaps, diabetes induction occurred 3 days after surgery, and they were sacrificed after 7 weeks. Diabetic rats with 10 mm gaps demonstrated an impaired ability to form bridging cables, the initial step of regeneration through chambers. Morphometric studies of bridging cables between transected nerve ends demonstrated a significant reduction in the mean endoneurial area in diabetic animals with 5 and 8 mm gaps compared to controls. The number of regenerated myelinated axons in the chamber was significantly decreased in diabetic rats with 8 and 10 mm gaps. The mean myelinated fiber area in the regenerated cables of the diabetic group was significantly decreased with 5 mm gaps and significantly increased with 8 mm gaps compared to controls. Size-frequency histograms of regenerated myelinated fiber areas suggest a delay in the maturation of small caliber axons. Schwann cell migration across 5 mm gaps was examined with S-100 immunohistochemistry. The total distance of Schwann cell migration into cables from both proximal and distal ends was significantly reduced in diabetic animals. Characterization of PNR across gaps through silicone chambers in diabetic rats showed impairment in multiple aspects of the regenerative process, including cable formation, Schwann cell migration, and axonal regeneration. PMID- 9219864 TI - Involvement of glutathione peroxidase and catalase in the disposal of exogenous hydrogen peroxide by cultured astroglial cells. AB - The ability of astroglial cells to detoxify exogenously applied hydrogen peroxide (H2O2) was tested using astroglia-rich primary cultures derived from the brains of newborn rats. Incubation of astroglial cells with 100 microM H2O2 in the absence of glucose led to a 66% oxidation of the cellular glutathione within 30 s. Under these conditions, the cells were unable to re-establish the original high ratio of GSH/GSSG within 30 min of incubation. In contrast, if glucose was present the amount of GSSG produced on incubation with H2O2 was smaller (45% of total glutathione after 30 s) and the original ratio of GSH/GSSG was almost completely re-established within 10 min. If 100 microM H2O2 was applied, H2O2 disappeared from the incubation buffer with an apparent half-life of approximately 4 min. After 15 min of incubation, no H2O2 was detectable any more. The apparent half-life of H2O2 in the incubation buffer increased slightly but significantly with increasing concentration of H2O2 or when the cells were starved of glucose. A small reduction in the capacity of the cells to detoxify H2O2 was also observed after depletion of the glutathione content to 14% of control level by a 24 h pre-incubation of the cells in culture medium containing buthionine sulfoximine, an inhibitor of glutathione synthesis. Incubation of astroglial cells with mercaptosuccinate or 3-aminotriazole, inhibitors of glutathione peroxidase and catalase, respectively, only marginally reduced the rate of disappearance of H2O2 from the incubation buffer. In contrast, the rate of H2O2 clearance was strongly reduced in the presence of both inhibitors. These results demonstrate that glutathione peroxidase and catalase are involved in the detoxification of H2O2 by astroglial cells and that both enzymes are able to substitute for each other in the detoxification of H2O2. PMID- 9219865 TI - The effects of type I and type II corticosteroid receptor agonists on exploratory behavior and spatial memory in the Y-maze. AB - We investigated the effects of two adrenal steroid agonists on adrenalectomized (ADX) rats' performance on the Y-maze. The Y-maze was chosen because memory can be assessed quickly and because it is sensitive to various parameters of exploratory behavior and spatial memory performance. Four days after surgery, ADX rats were injected with aldosterone (ALDO, a selective Type I receptor agonist), RU362 (a selective Type II receptor agonist) or sesame vehicle at three different time points (120 min prior to Trial 1, immediately after Trial 1 or 120 min after Trial 1). SHAM-operated rats injected with vehicle were also tested. The results indicate that vehicle-treated ADX rats were impaired on spatial recognition memory compared to SHAM rats. Treatment with ALDO restored spatial recognition memory performance of ADX rats to a level comparable to SHAM-treated rats by acting on acquisition and consolidation, whereas treatment with RU362 did not change the poor spatial recognition memory performance of ADX rats. Discrimination memory was improved only when either agonist was injected prior to the first trial, strongly suggesting a non-selective effect of corticosteroids on discrimination memory such as increasing arousal. A detailed analysis of exploratory behavior showed that both the ALDO- and RU362-treated rats explored the Y-maze more than the ADX and SHAM groups at all periods of the experiment. These results show that the non-specific increase in exploratory behavior induced by replacing corticosteroids targeted at Type I and Type II receptors was used differentially with the ALDO-treated rats learning and consolidating spatial information better than the RU362-treated rats. These data are discussed along with other evidence to suggest that Type II receptors may require the simultaneous occupancy of Type I receptors to affect learning and memory processes. PMID- 9219866 TI - Human sleep EEG following the 5-HT1A antagonist pindolol: possible disinhibition of raphe neuron activity. AB - The sleep electroencephalogram (EEG) was used to assay central effects of pindolol (10 and 30 mg p.o.), a mixed beta(1/2)-adrenoceptor/5-hydroxytryptamine (5-HT)(1A/1B) receptor blocker, in humans. Compared to placebo, pindolol produced a dose-related suppression of rapid-eye-movement (REM) sleep, including a prolongation of REM latency, and a decrease of REM time and REM density. At the higher dose, it also reduced EEG spectral power during non-REM sleep in portions of the delta, theta, and alpha frequencies (1.125-5.125 Hz, 7.125-9.625 Hz). By contrast, betaxolol (20 mg p.o.), a selective beta1-antagonist devoid of serotonergic affinity, affected neither REM sleep nor EEG power. REM sleep is, in part, under the inhibitory control of serotonergic neurons projecting from the dorsal raphe nucleus to pontine cholinergic/cholinoceptive cells. The EEG power spectrum induced by pindolol tended to be opposite to what has previously been reported for ipsapirone, a 5-HT1A agonist. Therefore, the present data, tentatively, are consistent with the contention that pindolol inhibits, possibly selectively, somatodendritic 5-HT1A autoreceptors in humans and may antagonize self-inhibition of midbrain raphe nuclei 5-HT neurons. PMID- 9219868 TI - New therapeutic possibility of blocking cytokine-induced neutrophil chemoattractant on transient ischemic brain damage in rats. AB - Earlier we indicated that neutrophilic invasion into cerebral parenchyma is an important step in rat cerebral ischemia-reperfusion injury and the production of chemotactic factors, cytokine-induced neutrophil chemoattractant (CINC) precede the neutrophilic invasion. The aim of the present study was to evaluate the role of CINC production and the therapeutic possibility of blocking CINC activity in the transient ischemic brain damage in rats. Focal transient ischemia was produced by intraluminal occlusion of the right middle cerebral artery for 60 min. An enzyme immunoassay was used to measure the brain concentration of CINC and myeloperoxidase activity in ischemic areas was measured as a marker of neutrophilic accumulation. An immunohistochemical staining technique was used to detect the immunopositive cells for anti-CINC antibody. Further, application of anti-CINC antibody or anti-neutrophil antibody to rats was used to evaluate the role of CINC production. In ischemic areas, CINC production was detected and peaked 12 h after reperfusion, which followed 60 min of ischemia. Intraperitoneal injection of anti-neutrophil antibody 24 h before and immediately after reperfusion significantly reduced the brain water content and partially reduced the CINC production in ischemic areas. Further, immunohistochemical staining showed that anti-CINC antibody was found on the endothelial surface of venules and on parts of neutrophils that had invaded the ischemic area 6 to 24 h after reperfusion. Also, treatment with anti-CINC antibody reduced ischemic edema formation 24 h after reperfusion and the size of infarction areas 7 days after reperfusion. It thus appears that CINC, mainly produced by endothelium activated by factors released from neutrophils, plays an important role in ischemic brain damage. Furthermore, the blocking of CINC activity with antibody suggests an immuno-therapeutic approach to the treatment of stroke patients. PMID- 9219869 TI - Phasic relationship between the activity of basal forebrain neurons and cortical EEG in urethane-anesthetized rat. AB - Previous studies have shown that a large number of neurons in the basal forebrain have higher firing rates when the cortical electroencephalogram (EEG) is characterized by low-voltage fast activity compared to states characterized by slow waves. A smaller number of cells with increased discharge rates during slow waves have also been observed. This putative ascending effect is thought to be tonic, but no attempt has been made to analyze a closer temporal correlation between the activity of basal forebrain neurons and the cortical EEG. Recordings were made from single units in the basal forebrain concurrently with the cortical EEG in urethane-anesthetized rats. A total of 52 neurons consistently showed higher firing during low-voltage fast activity (F-cells), whereas 14 neurons were consistently more active during cortical slow waves (S-cells). In most of the F- (90%) and S-cells (86%) the change in firing rate occurred prior to the change in the EEG. The average delay was 300-400 ms. At a deep level of anesthesia, the EEG was characterized by an alternation of flat periods and large waves. Most F-cells became active near the start of the first large wave, which is known to correspond to the onset of depolarization of cortical pyramidal neurons. In contrast, most S-cells were less active during the large waves. These data show that the activity of basal forebrain neurons is phasically correlated with the EEG in addition to the tonic correlation that has been demonstrated previously. Both types of basal forebrain neurons change their firing rate prior to the change in cortical EEG, suggesting that the basal forebrain neurons may have a regulatory influence on the EEG. PMID- 9219867 TI - Homeostasis of ocular surface epithelium in the rat is regulated by opioid growth factor. AB - Endogenous opioid peptides serve as growth factors in developing, renewing, and neoplastic cells and tissues. This study examined the hypothesis that opioids serve to modulate the homeostatic renewal of ocular surface epithelium in the rat. DNA synthesis in the epithelium of the central (CC) and peripheral (PC) cornea, limbus (LM), and conjunctiva (CN) was investigated using adult male rats. Animals received an injection of opioid growth factor (OGF), [Met5]-enkephalin, OGF and naloxone (NAL), NAL alone, naltrexone (NTX), or an equivalent volume of sterile water (CO) and sacrificed 4 h later (i.e. 16:00 h). [3H]thymidine was administered 1 h before sacrifice. With the exception of NTX (20 mg/kg), all compounds were given at 10 mg/kg. Examination of 5 time points over an 18-h period revealed no variation in DNA synthesis within a region of ocular surface basal epithelium (BE). OGF depressed DNA synthesis of the BE by 25, 48, and 50% in the PC, LM, and CN, respectively; little labeling was recorded in the BE of the CC. Exposure to OGF-NAL or NAL alone did not alter DNA synthesis of the BE. Complete blockade of OGF-zeta receptor interaction by administration of the potent opioid antagonist, NTX, increased the number of epithelial cells in the PC, LM, and CN undergoing DNA synthesis by 30 to 72%. The effects of OGF and NTX on DNA synthesis of BE also were observed in an organ culture setting. Utilizing immunocytochemistry, OGF and its receptor zeta were associated with both the basal and the suprabasal cells of the ocular surface epithelium. These results indicate that an endogenous opioid peptide, OGF, and its receptor are present and govern homeostatic cellular renewal processes in ocular surface epithelium. OGF regulates DNA synthesis in a direct manner, and does so by a tonic, inhibitory, and receptor-mediated mechanism. PMID- 9219870 TI - The avian somatosensory system: the pathway from wing to Wulst in a passerine (Chloris chloris). AB - The organization of the wing component of the dorsal column-medial lemniscal pathway, and somatosensory projections from the thalamus to the Wulst, are described for an oscine member of the major group of birds, the Passeriformes. Wing primary afferents terminate throughout the cervical spinal cord, but between the brachial enlargement and the spino-medullary junction, they are confined to medial lamina V. Within the medulla, terminations extend rostrally and laterally to occupy the cuneate (Cu) and external cuneate nuclei (CuE). Ascending projections from Cu and CuE form the contralateral medial lemniscus, which has extensive projections to the midbrain and to the thalamus. In the midbrain the projections surround the central auditory nucleus densely, and terminate more sparsely within it. In the thalamus, specific terminations were observed in nucleus uvaeformis and in the nucleus dorsalis intermedius ventralis anterior (DIVA). DIVA projects to the ipsilateral rostral Wulst where it terminates in the intercalated hyperstriatum accessorium, in a distinct, regular patchy fashion. The somatosensory projections to the telencephalon in green finch are similar to those in pigeon, but dissimilar to those in budgerigar. PMID- 9219871 TI - Methamphetamine exposure can produce neuronal degeneration in mouse hippocampal remnants. AB - Neuronal cell death in hippocampal remnants was seen after methamphetamine (METH) exposure. Two techniques (Fluoro-Jade labeling and argyrophylia) showed that neuronal degeneration occurred in the indusium griseum, tenia tecta and fasciola cinerea within 5 days post-METH exposure in 70% of the mice. Neurodegeneration also occasionally occurred in the piriform cortex, hippocampus and frontal/parietal cortex. This cell death, unlike striatal neurotoxicity, was not dependent on magnitude of hyperthermia occurring but did correlate with behavioral seizure activity during METH exposure. Excitotoxic mechanisms may be underlying the neuronal degeneration since co-administration of phenobarbital blocked cell death. PMID- 9219872 TI - Recovery of taste aversion learning induced by fetal neocortex grafts: correlation with in vivo extracellular acetylcholine. AB - Rats showing disrupted taste aversion due to insular cortex lesions, received either homotopic or heterotopic (occipital) cortical fetal brain grafts. Behavioral results showed that the recovery of the ability to acquire conditioned taste aversions induced by fetal grafts depended on post-graft time (45 but not at 15 days) and tissue specificity (homotopic but not heterotopic). In vivo analysis of acetylcholine (ACh) release revealed that only the group receiving homotopic grafts and tested 45 days post graft had a release of ACh after KCl stimulation similar to that in the control group. Furthermore, homotopic grafts and lesioned groups showed significantly weaker specific receptor binding of [3H]L-glutamate compared with controls. These results suggest that ACh is specifically involved in the process of behavioral recovery induced by homotopic cortical transplants. PMID- 9219873 TI - Mutations in the GAP-related domain impair the ability of neurofibromin to associate with microtubules. AB - The neurofibromatosis 1 (NF1) gene encodes a cytoplasmic protein with structural and functional homology to GTPase activating proteins (GAPs) for p21-ras. Double labeling immunofluorescence experiments using neurofibromin antibodies and in vitro microtubule assembly have demonstrated that the NF1 gene product, neurofibromin, interacts with cytoplasmic microtubules. The region critical for this interaction was shown to reside within the NF1-GAP-related domain (NF1GRD). Examination of the NFIGRD reveals a number of residues that are highly conserved amongst all GAP molecules. Mutational analysis of a representative number of these conserved residues demonstrated differential effects on NF1GRD p21-ras GAP activity. In this study, we examined the effect of these selected NF1GRD mutations on the ability of neurofibromin to associate with microtubules. Mutations at residues R1391, P1400 and K1423 disrupted microtubule association. In contrast, mutations at residues E1264, Q1426 and the insertion of exon 23a, critical for p21-ras regulation, did not impair microtubule association. These results demonstrate that some residues important for p21-ras regulation are also required for microtubule binding while other residues within the NF1GRD differentially affect p21-ras regulation and microtubule association. PMID- 9219874 TI - Genetic association between susceptibility to Parkinson's disease and alpha1 antichymotrypsin polymorphism. AB - The apolipoprotein E (ApoE*) gene is a major risk factor of developing Alzheimer's disease (AD) and the alpha1-antichymotrypsin (ACT) polymorphism is likely to modify susceptibility of the ApoE* gene for AD. Because pathogenesis of AD is partly similar to that in idiopathic Parkinson's disease (PD), we investigated the distribution of genotypes of the ApoE and the ACT in patients with PD. The number of individuals with two copies of the ACT-A allele (ACT-AA genotype) in patients with PD increased significantly compared to that in healthy controls (19.9% versus 8.3%, P < 0.02), and the ACT-A allele frequency in patients with PD was significantly higher than that in healthy controls (chi2 = 5.96, df = 1, P < 0.015). The odds ratio for developing PD in individuals with the ACT-AA genotype was 3.36 compared to individuals with two copies of another allele, the ACT-T allele (ACT-TT genotype). There was no association between ApoE genotypes and susceptibility to PD. These data suggest that the etiological basis of PD might be partly similar to that of AD and the ACT gene might be one of the susceptibility factors for PD. PMID- 9219875 TI - Serotonin1A receptor agonists induce Fos protein expression in the locus coeruleus of the conscious rat. AB - The azapirones, which are partial agonists of the serotonin (5-HT)1A receptor, possess anxiolytic activity. These agents may act at the pre- or postsynaptic 5 HT1A receptors, and involve the noradrenergic system. To determine whether these drugs activate noradrenergic neurons via 5-HT1A receptors, we have evaluated the expression of the immediate early gene c-fos in the locus coeruleus. Tandospirone and ipsapirone each induced expression of Fos protein in the noradrenergic neurons of the locus coeruleus of conscious rats. This effect was reversed by pretreatment with (+)-WAY100135, a specific 5-HT1A antagonist. These results clearly demonstrate that azapirones activate noradrenergic neurons via 5-HT1A receptors. PMID- 9219876 TI - OFF pathway is preferentially suppressed by the activation of GABA(A) receptors in carp retina. AB - This study examines the effect of gamma-aminobutyric acid (GABA) on the ON and OFF pathways in isolated, superfused carp retina. In most (76%) of amacrine cells bath-applied GABA preferentially suppressed the OFF response. The effect of GABA was blocked by bicuculline. Baclofen did not cause a similar effect. Furthermore, GABA produced a substantial suppression of the OFF bipolar cell response. We conclude that the preferential suppression of the OFF pathway may be due to a presynaptic inhibition mediated via GABA(A) receptors at the terminals of OFF bipolar cells. PMID- 9219877 TI - Astrocytes in the aged rat spinal cord fail to increase GFAP mRNA following sciatic nerve axotomy. AB - Aging in the brain is associated with specific changes in the astrocyte population. The present study establishes that similar changes occur in the aging spinal cord. The levels of glial fibrillary acidic protein (GFAP) mRNA were significantly increased 0.4-fold in aged 8- to 17-month-old rats compared to young 2-month-old rats. The ability of astrocytes in the aging spinal cord to respond to a non-invasive CNS injury was compared to young rats 4 days following sciatic nerve axotomy. The level of GFAP mRNA was significantly increased 0.5 fold in the young rats in response to axotomy. In contrast, the level of GFAP mRNA in aged rats did not increase following injury above that present in non axotomized rats of the same age. PMID- 9219878 TI - Exposure to long summer days affects the human melatonin and cortisol rhythms. AB - Exposure of 8 human subjects in summer to a natural 16 h bright light photoperiod phase advanced the morning salivary melatonin decline and cortisol rise and shortened the nocturnal melatonin signal by 2 h relative to the winter patterns of the same subjects followed under a combined artificial and natural light 16 h photoperiod. The data suggest that summer days experienced from sunrise till sunset and not winter days with a combined artificial and natural light long photoperiod evoke a true long day response of the human circadian system. PMID- 9219879 TI - PAG-microinjected dipyrone (metamizol) inhibits responses of spinal dorsal horn neurons to natural noxious stimulation in rats. AB - In addition to their well-known peripheral and spinal effects, non-steroidal antiinflammatory drugs (NSAIDs) are believed to diminish nociceptive responses by acting supraspinally and activating descending modulatory systems. We have herein investigated whether this descending action involves a depression of spinal sensory neurons. In rats under barbiturate anesthesia, responses of lumbar wide dynamic-range neurons to a noxious clamp in their receptive fields were depressed to 46% of baseline value by the microinjection of 100 microg dipyrone (metamizol) into the periaqueductal gray matter (PAG). These results show that PAG application of NSAIDs activates descending systems which depress the excitation of spinal sensory neurons by natural noxious stimuli. PMID- 9219880 TI - Substance P analogues potentiate the pressor response to microinjection of L glutamate into laminas I and II of the cat dorsal horn. AB - Microinjection of a substance P analogue (1 mM; 7 or 10 nl) into laminae I and II of the L7 dorsal horn of decerebrate cats significantly potentiated (P < 0.05) the increase in arterial pressure evoked by microinjection of L-glutamate (109 mM; 7 or 10 nl) into these spinal sites. Microinjection of the substance P analogues (i.e., GR73638 and [Sar9,Met(O2)11]-substance P) which were selective NK-1 receptor agonists, had no impact on the cardioacceleration evoked by microinjection of L-glutamate (P > 0.05). In addition, microinjection of these analogues had no effect on the modest and non-significant increase in phrenic nerve discharge evoked by L-glutamate. We conclude that stimulation of NK-1 receptors in the superficial laminae of the dorsal horn potentiates the pressor responses to microinjection of L-glutamate. PMID- 9219881 TI - Techniques and mechanisms of action of transcranial stimulation of the human motor cortex. AB - Electrical and magnetic methods are available to stimulate the human brain through the intact scalp. Although both are successful, magnetic stimulation is now used almost exclusively because the discomfort is minimal compared with that caused by electrical stimulation. Nevertheless, electrical stimulation is still used occasionally since comparison of results from both techniques can often yield useful clinical and scientific information not available from either method in isolation. PMID- 9219882 TI - The methodology and scope of human microneurography. AB - Microneurography was introduced in 1967 and has developed into an invaluable tool for investigating human somatosensory, motor and cardiovascular physiology and pathophysiology. It involves percutaneous insertion of a metal microelectrode into fascicles of limb and facial nerves. This review covers the procedures and equipment necessary for microneurography and provides a current circuit for a preamplifier. Evidence is presented that (i) most recordings from myelinated axons involve an effective penetration of the myelin by the electrode; (ii) based on physiological criteria, microstimulation through the electrode can be used to activate single axons although the probability of this is relatively low and (iii) despite 'micro' lesions caused by the electrode insertion into the nerve and its fascicles, the morbidity with the procedure is acceptably low. PMID- 9219883 TI - Unit identification, sampling bias and technical issues in microneurographic recordings from muscle spindle afferents. AB - Recordings have been made directly from human muscle spindle afferents for some 3 decades, and these have allowed assessment of fusimotor function in human subjects during normal motor acts and in patients with motor disturbances. However, inferences about fusimotor function are indirect, valid only if identification of the axon as of muscle spindle origin is secure and all extrafusal influences on the spindle have been controlled. As is discussed, the identification of spindle afferents and their classification into Group Ia and Group II are more problematic than in the cat, but these problems are probably relatively minor given the insight into normal function that can come from appropriately designed experiments in awake cooperative human subjects. PMID- 9219884 TI - The study of normal and abnormal neuromuscular transmission with single fibre electromyography. AB - The use of single fibre electromyography (SFEMG) in the study of neuromuscular transmission across individual motor endplates in situ is reviewed. The neuromuscular jitter can be studied both during voluntary contraction and electrical activation of the muscle fibre. The differences, pitfalls and advantages of these methods are discussed. Findings in myasthenia gravis and other disorders of the neuromuscular transmission are examined. PMID- 9219885 TI - Spike train analysis. AB - Procedures for the analysis of stimulus-correlated spike train data are reviewed. All procedures considered attempt to extract excitability changes evoked by the stimulus at the neuron investigated. The methods covered range from rather simple methods that require very little computational effort (raw spike train displays; peri-stimulus-time histogram (PSTH)) to more sophisticated procedures that attempt to extract all information available in the recorded spike-train data. PMID- 9219886 TI - Estimating post-synaptic potentials in tonically discharging human motoneurons. AB - The activity of single motor units in human muscles can be recorded with relative ease, and the spike train of a single motor unit precisely reflects the spike train of the parent motoneurone. This has led to the proposal of a number of methods to estimate stimulus-evoked post-synaptic potentials in human motoneurones. All of these methods rely on manipulating the spike trains of motor units over a number of trials. All are based on a number of assumptions, all have limitations, and none so far have passed the test of a direct comparison of the estimate of the shape of the post-synaptic potential with a direct intracellular measurement of it. These techniques are summarised in this review. PMID- 9219887 TI - A review of recent applications of cross-correlation methodologies to human motor unit recording. AB - This article reviews some recent applications of time and frequency domain cross correlation techniques to human motor unit recording. These techniques may be used to examine the pre-synaptic mechanisms involved in control of motoneuron activity during on-going motor tasks in man without the need for imposed and artificial perturbations of the system. In this review we examine, through several examples, areas in which insights have been gained into the basic neurophysiological processes that bring about motoneuron firing in man and illustrate how these processes are affected by central nervous system pathology. We will demonstrate that synchronization and coherence may be revealed between human motor unit discharges and give examples that support the hypothesis that these phenomena are generated by activity in a focused common corticospinal input to spinal motoneurons. Disruption of central motor pathways due to diseases of the nervous system leads to pathophysiological alterations in the activity of these pre-synaptic motoneuron inputs that can be revealed by cross-correlation analysis of motor unit discharges. The significance of these studies and outstanding questions in this field are discussed. PMID- 9219888 TI - Assessing changes in presynaptic inhibition of Ia afferents during movement in humans. AB - Different methods, based on different principles, have been proposed to estimate changes in presynaptic inhibition of Ia terminals (accompanied by primary afferent depolarization, (PAD)) during voluntary contraction in humans. (i) A discrepancy between the H-reflex amplitude, at an equal level of EMG activity, in two situations (e.g., walking and standing) may be taken as suggesting a different control of PAD interneurones in the two cases. (ii) A conditioning stimulation (vibration or electrical stimulation) is used to activate PAD interneurones and to evoke presynaptic inhibition of the afferent volley of the test reflex. The resulting long-lasting depression of the reflex depends on the excitability of PAD interneurones, but can be contaminated by long-lasting post synaptic effects. (iii) The amount of reflex facilitation evoked by a purely monosynaptic Ia volley varies inversely with the on-going presynaptic inhibition of Ia afferents mediating the conditioning volley, and can be used to assess this on-going presynaptic inhibition. None of these methods can provide by itself unequivocal evidence for a change in presynaptic inhibition of Ia terminals, but reasonably reliable interpretations may be proposed when congruent results are obtained with different methods. Thus it has been shown that, during selective voluntary contraction, presynaptic inhibition is decreased on Ia afferents projecting on motoneurones of the contracting muscle and increased on Ia afferents projecting on motor nuclei not involved in the contraction. PMID- 9219889 TI - Neurophysiological methods for studies of the motor system in freely moving human subjects. AB - In this paper, the following experimental methods for studies of the motor system in freely moving human subjects will be considered: (i) eliciting the H-reflex and understanding its use as a test response, (ii) methods to measure reciprocal inhibition between antagonist muscles, (iii) methods to measure presynaptic inhibition of Ia-afferent terminals in the spinal cord, (iv) certain aspects of the interpretation of peri-stimulus time histograms (PSTH) of single motor unit discharge, and finally, (v) stimulation of the motor cortex and the measurement of response parameters that may reflect task dependent changes. Two closely related ideas bearing directly on these methods will be emphasized--the influence of the background level of motor activity on input output properties of the neural pathway investigated and the operating point on the input-output curves at which the experimental variable is measured. Finally, in the discussion a simple model that is easily understandable in geometric terms is presented to help predict and interpret the outcome of these sorts of experiments. PMID- 9219890 TI - Measurement of human muscle fatigue. AB - Human muscle fatigue has been studied using a wide variety of exercise models, protocols and assessment methods. Based on the definition of fatigue as 'any reduction in the maximal capacity to generate force or power output', the different methods to measure fatigue are discussed. It is argued that reliable and valid measures must include either assessment of maximal voluntary contraction force or power, or the force generated by electrical stimulation. By comparing tetanic stimulation and maximal voluntary contraction force one may reveal whether fatigue is of central origin, or whether peripheral mechanisms are involved. Adequate use of twitch interpolation provides an even more sensitive measure for central fatigue. Indirect methods as endurance times and electromyography show variable responses during exercise and no close relationship to fatigue. Hence these methods are of limited value in measurement of human muscle fatigue. PMID- 9219891 TI - Functional magnetic resonance imaging of the human brain. AB - The current technical and methodological status of functional magnetic resonance imaging (fMRI) is reviewed. The mechanisms underlying the effects of deoxyhemoglobin concentration and cerebral blood flow changes are discussed, and methods for monitoring these changes are described and compared. Methods for post processing fMRI data are outlined. Potential problems and solutions related to vessels and motion are discussed in detail. PMID- 9219892 TI - Images of the working brain: understanding human brain function with positron emission tomography. AB - In the past 15 years positron emission tomography (PET) has become a settled method of imaging the functioning human brain, both in normal volunteers and in patients with various disorders. Much of the work on sensory systems has been on the visual system, a conveniently studied and very important part of the brain. The motor system in health and disease has attracted as much interest. In this short review I will explain some of the technical aspects of PET, and illustrate some of the research that has been done on visual and motor brain function in the human. PMID- 9219893 TI - Immunological identification of cholesterol ester hydrolase in the steroidogenic tissues, adrenal glands and testis. AB - Monoclonal antibodies were generated against the purified pancreatic cholesterol ester hydrolase (CEH, EC 3.1.1.13) to examine the expression of CEH in various bovine tissues. The presence of CEH isozyme antigenically indistinguishable from pancreatic enzyme in the steroidogenic tissues, adrenal glands and testis has been first demonstrated here using the immunoprecipitation method. These results suggest that CEH isozyme, similar to pancreatic CEH, might be involved in the cholesterol metabolism in the steroidogenic tissue. PMID- 9219895 TI - Regulation of platelet-activating factor (PAF) biosynthesis via coenzyme A independent transacylase in the macrophage cell line IC-21 stimulated with lipopolysaccharide. AB - The regulation of PAF synthesis by the macrophage cell line IC-21 challenged with bacterial endotoxin was investigated. The LPS-induced increase in cellular PAF levels was rapid, sustained and attained maximal levels within 30 min following LPS stimulation. PAF accumulation was accompanied by the activation of the CoA independent transacylase and acetyl-CoA: lyso-PAF acetyltransferase, whereas the release of free [3H]arachidonic acid in prelabeled cells reflecting the activation of phospholipase A2, occurred primarily within the initial 1-5 min of treatment with LPS. Cell lysates from LPS-stimulated macrophages exhibited a markedly increased enzymatic activity that was capable of both acylation of 1 [3H]alkyl-2-lyso-GPC (lyso-PAF) and deacylation of 1-[3H]alkyl-2-acyl-GPC generating [3H]lyso-PAF via CoA-independent transacylation of exogenous lysoplasmenylethanolamine compared with extracts from resting macrophages. Pretreatment of the cells with LPS for 5 and 30 min enhanced significantly the transfer of [14C]arachidonic acid from 1-[3H]alkyl-2-[14C]arachidonoyl-GPC into plasmenylethanolamine in prelabeled cell homogenates following the addition of exogenous lysoplasmenylethanolamine. Taken together, these data suggest that the CoA-independent transacylase, but not phospholipase A2, is a key enzyme responsible for the prolonged generation of lyso-PAF and that the increased capability of CoA-independent transacylation followed by CoA-dependent acetylation of lyso-PAF can sustain the biosynthesis of PAF in LPS-stimulated IC 21 macrophages. PMID- 9219894 TI - Discovery of 5R-lipoxygenase activity in oocytes of the surf clam, Spisula solidissima. AB - Arachidonic acid and 5-hydroxyeicosatetraenoic acid (5-HETE) are reported to induce reinitiation of meiosis in oocytes of the surf clam Spisula sachalinensis from the Sea of Japan (Varaksin et al., Comp. Biochem. Physiol. 101C, 627-630 (1992). As the Atlantic surf clam Spisula solidissima is a commonly used model for the study of meiosis reinitiation, we examined these cells for the possible occurrence of lipoxygenases and for the bioactivity of the products. Incubation of [14C]arachidonic acid with homogenates of S. solidissima oocytes led to the formation of two major metabolites: 5R-HETE, a novel lipoxygenase product, and 8R HETE. The products were identified by HPLC, uv spectroscopy, and GC-MS. The corresponding hydroperoxy fatty acids, the primary lipoxygenase products, were isolated from incubations of ammonium sulfate fractionated oocyte cytosol. Arachidonic and eicosapentaenoic acids were identified as constituents of S. solidissima oocyte lipids and the free acids were equally good lipoxygenase substrates. We examined the activity of C18 and C20 polyunsaturated fatty acids and their lipoxygenase products on meiosis reinitiation in Spisula solidissima oocytes, using serotonin and ionophore A23187 as positive controls. The fatty acids and their derivatives were inactive. We conclude that in the surf clam, (as in starfish), there are responding and non-responding species in regard to the maturation-inducing activity of the oocyte lipoxygenase products, and that the lipoxygenase has another, as yet uncharacterized, function in oocyte physiology. PMID- 9219896 TI - Structural organization of lipid phase and protein-lipid interface in apolipoprotein-phospholipid recombinants: influence of cholesterol. AB - The complexes of individual human plasma apolipoproteins (apo) A-I, E and A-II with dipalmitoylphosphatidylcholine (DPPC) in the absence or in the presence of cholesterol (Chol) were prepared with initial DPPC/Chol/protein weight ratio as 3:0.15:1. ApoA-I/DPPC/Chol complexes with different protein content (initial DPPC/apoA-I weight ratios were changed from 10.5:1 to 2.6:1) but with a fixed initial DPPC/Chol weight ratio of 20:1 were also prepared. The complexes were isolated by gel-filtration and characterized by size and composition. ApoA-I- and apoA-II-complexes had the same size (80-84 A) and the complexes became more heterogeneous upon Chol inclusion; apoE-complexes were larger (97-100 A) and more homogeneous and Chol addition had no effect on their hydrodynamic properties. Chol seems to be excluded partially in the following manner for isolated complexes with different apo's: A-II > E > A-I. The possible existence of two lipid regions in the complexes differing in lipid dynamics - the lipid shell in the vicinity of apolipoprotein (boundary lipid) opposite to the remaining part of the lipid bilayer - has been studied by absorbance and fluorescence spectroscopy with cis-parinaric acid (cis-PA) and trans-parinaric acid (trans-PA) embedded into the complexes. Their application is based on a strong preference of trans-PA for solid lipid while cis-PA distributes more equally between co-existing fluid and solid lipid regions (Sklar et al. (1979) Biochemistry 18, 1707-1716). (1) For apoA-I-complexes, the partition of cis-PA between water and lipid phase at temperatures below and above the transition temperature of DPPC (T(t)) was insensitive to Chol and temperature, while partition of trans-PA into the lipid phase of Chol-containing complex was increased at high temperature and decreased at low temperature. These results seem to be related to trans-PA redistribution between Chol-rich and protein-rich lipid domains, the latter being more disordered at T < T(t) and more immobilized at T > T(t) compared to the bulk bilayer; cis-PA localizes preferentially in boundary lipid. This hypothesis was directly confirmed by measurements of energy transfer between apoA-I tryptophanyls and probe molecules. (2) The relative response of trans-PA fluorescence intensity to temperature-induced phase transition of DPPC in apoA I/DPPC/Chol complexes was decreased as a function of apolipoprotein content in a non-monotonic fashion with a transition midpoint at a mol ratio DPPC/A-I of 250:1, probably indicating two different modes of apolipoprotein/DPPC interaction in different sized complexes. (3) The comparative study of lipid dynamics in apoA I-, apoE- and apoA-II-containing complexes with temperature response to phospholipid phase transition with fluorescence parameters such as intensity and anisotropy of cis-PA and trans-PA revealed the presence of boundary lipid in all three complexes without Chol. In contrast to apoA-I-containing complexes, in apoA II/DPPC/Chol complexes, trans-PA seems to move preferentially into boundary lipid and cis-PA to distribute between two different regions probably as a result of more ordering action induced by apoA-II compared to apoA-I on the nearest phospholipid molecules in Chol-containing complexes; the apoE action on trans-PA and cis-PA distribution could be intermediate. Based on these results, the degree of Chol exclusion from the boundary lipid region for complexes with different apo's increasing in the order A-II > E > A-I can be suggested. Different Chol distributions between two lipid regions in the complexes seems not to be a function of complex size, but rather is an inherent property of the particular apolipoprotein molecule. PMID- 9219897 TI - In vivo metabolism of 24R,25-dihydroxyvitamin D3: structure of its major bile metabolite. AB - In vivo metabolism of 24R,25-dihydroxyvitamin D3 (24,25-(OH)2D3) in female dogs has been studied thoroughly, and its major bile metabolite identified. After single oral administration of 24,25-(OH)2 [6,19,19-3H]D3 the plasma concentrations of radioactive metabolites were monitored for 504 h, and the metabolites in the bile collected and analyzed. The concentration of 24,25 (OH)2D3 in plasma reached a maximum after 6 h and decayed in two distinct phases; a fast-phase with a half-life of 17 h, followed by a slow-phase with a 17-day half-life. The area under the concentration/time curve (AUC) was 78-84% (0-504 h). The only detectable metabolite in the plasma was 25-hydroxy-24-oxovitamin D3 whose AUC was less than 5%. At 504 h, about 50% of administered radioactivity has been excreted, of which about 90% was found in the feces, indicating most of the administered 24,25-(OH)2D3 to be excreted in bile. A major metabolite, which constituted 23% of the total bile radioactivity at 504 h, was found in the bile. This metabolite was efficiently deconjugated by beta-glucuronidase to afford an aglycone which was identified as 23S,25-dihydroxy-24-oxovitamin D3 (23S,25-(OH)2 24-oxo-D3), by co-chromatography on HPLC with synthetic standards. The glucuronide was isolated from the bile of dogs given large doses of 24,25 (OH)2D3, and the structure determined being 23-(beta-glucuronide) of 23S,25-(OH)2 24-oxo-D3, by analyzing its negative ion mass spectrum and the positive ion mass spectrum of its derivatives. Thus it was concluded that, in dogs, 24,25-(OH)2D3 is a long lasting vitamin D metabolite, is mainly excreted in bile when metabolized to 23S,25-(OH)2-24-oxo-D3 and is conjugated at 23-OH as glucuronide. PMID- 9219898 TI - A thermodynamic analysis of the partitioning of cholesterol and related compounds between trioleoylglycerol and egg phosphatidylcholine bilayers. AB - The free energy of transfer of a number of alcohols, including cholesterol, from a bulk isotropic lipid phase, trioleoylglycerol (TG), to an anisotropic lipid phase, egg phosphatidylcholine (PC), was determined. n-Alkane-1-ols partitioned preferentially into the bilayer phase; for example, the free energy of transfer of octanol-1 from TG to PC was about -1.0 kcal/mol. This preference declined with increasing number of carbons at a rate of 40 cal/mol of CH2. Cholesterol had a much stronger preference for the bilayer with a free energy of -1.3 kcal/mol, compared to an extrapolated value of -0.2 kcal/mol for a normal alkane-1-ol with the same number of carbon atoms. Thus, the excess free energy of -1.1 kcal/mol represents the favourable interaction of the cholesterol skeleton with the bilayer phase. This conclusion was confirmed by comparing cholesterol 3 hemisuccinate to oleic acid. Substituting TG for water as the standard state has eliminated the large hydrophobic effect and has permitted us to identify for the first time the subtle binding increment of the steroid ring system. PMID- 9219899 TI - Acyl-CoA binding protein (ACBP) regulates acyl-CoA:cholesterol acyltransferase (ACAT) in human mononuclear phagocytes. AB - It is demonstrated that the acyl-CoA:cholesterol acyltransferase (ACAT) enzyme activity in rough endoplasmatic reticulum membranes is regulated by the acyl-CoA binding protein (ACBP). The ACAT activity is strongly inhibited by different ACBP/oleoyl-CoA complexes depending from the molar ratio of protein and fatty acid-CoA. Other lipid binding proteins such as bovine serum albumin and the liver fatty acid binding protein do not show any effects on ACAT activity. In addition, we can show that cholesterol loading with acetylated low density lipoproteins does not lead to an increase of the ACBP mRNA level. Consequently, the increase of the intracellular concentration of fatty acids because of the cholesteryl ester accumulation renders ACAT more active for cholesterol esterification. In binding studies we have characterized binding sites on microsomal membranes for the ACAT substrate oleoyl-CoA and the ACAT inhibitor diazepam. Diazepam competes with oleoyl-CoA and vice versa for its binding to microsomal membranes. This common binding site is suggested to be responsible for the transfer from ACBP bound oleoyl-CoA to ACAT and, therefore, to be essential for the microsomal cholesterol esterification. PMID- 9219900 TI - Evidence that 85 kDa phospholipase A2 is not linked to CoA-independent transacylase-mediated production of platelet-activating factor in human monocytes. AB - Platelet-activating factor (PAF) production is carefully controlled in inflammatory cells. The specific removal of arachidonate (AA) from 1-O-alkyl-2 arachidonoyl-sn-glycero-3-phosphocholine (GPC), thought to be mediated by CoA independent transacylase (CoA-IT), is required to generate the PAF precursor 1-O alkyl-2-lyso-GPC in human neutrophils. Exposure of A23187-stimulated human monocytes to the CoA-IT inhibitors SK&F 98625 and SK&F 45905 inhibited PAF formation (IC50s of 10 and 12 microM, respectively), indicating that these cells also need CoA-IT activity for PAF production. Because CoA-IT activity transfers arachidonate to a 2-lyso phospholipid substrate, its activity is obligated to an sn-2 acyl hydrolase to form the 2-lyso phospholipid substrate. SB 203347, an inhibitor of 14 kDa phospholipase A2 (PLA2), and AACOCF3, an inhibitor of 85 kDa PLA2, both inhibited AA release from A23187-stimulated human monocytes. However, AACOCF3 had no effect on A23187-induced PAF formation at concentrations as high as 3 microM. Further, depletion of 85 kDa PLA2 using antisense (SB 7111, 1 microM) had no effect on PAF production, indicating a lack of a role of 85 kDa PLA2 in PAF biosynthesis. Both SB 203347 and the 14 kDa PLA2 inhibitor scalaradial blocked PAF synthesis in monocytes (IC50s of 2 and 0.5 microM, respectively), suggesting a key role of 14 kDa PLA2 in this process. Further, A23187-stimulated monocytes produced two forms of PAF: 80% 1-O-alkyl-2-acetyl-GPC and 20% 1-acyl-2-acetyl-GPC, which were both equally inhibited by SB 203347. In contrast, inhibition of CoA-IT using SK&F 45905 (20 microM) had a greater effect on the production of 1-O-alkyl (-80%) than of 1-acyl (-14%) acetylated material. Finally, treatment of U937 cell membranes with exogenous human recombinant (rh) type II 14 kDa PLA2, but not rh 85 kDa PLA2, induced PAF production. Elimination of membrane CoA-IT activity by heat treatment impaired the ability of 14 kDa PLA2 to induce PAF formation. Taken together, these results suggest that a 14 kDa PLA2 like activity, and not 85 kDa PLA2, is coupled to monocyte CoA-IT-induced PAF production. PMID- 9219901 TI - Bacterial expression and characterization of human pancreatic phospholipase A2. AB - Mammalian pancreatic phospholipases A2 (PLA2) have recently been implicated in cell surface receptor-mediated inflammation. As a first step toward understanding how human pancreatic PLA2 (hp-PLA2) interacts with membranes and other biological targets including cell-surface receptors, we constructed its bacterial expression vector which can be used for the mutagenesis and protein over-expression. The expression vector (pSH-hp) was constructed using a synthetic hp-PLA2 gene whose transcription is controlled by T7 promoter. hp-PLA2 was expressed as a mature protein in high concentration in Escherichia coli cells and formed inclusion body. The solubilization of inclusion body protein followed by the refolding and purification produced ca. 5 mg of pure protein from one liter of growth medium. Kinetic studies of recombinant human, bovine and porcine pancreatic PLA2s using polymerized mixed liposomes and micelles as substrates showed that despite their highly homologous structures these mammalian pancreatic PLA2s have distinct phospholipid head group specificity and different activity toward various lipid substrates. PMID- 9219902 TI - Group II phospholipases A2 are indirectly cytolytic in the presence of exogenous phospholipid. AB - Systemic inflammatory response syndromes including septic shock and salicylate poisoning are associated with high circulating levels of secretory phospholipase A2 (sPLA2). In septic shock, sPLA2 has been implicated in the pathogenesis of multisystem organ failure, presumably by a direct cytotoxic effect on cells. The cytotoxicity of recombinant human sPLA2 and a venom PLA2 were examined on human erythrocytes, erythroleukemia cells and U937 cells. Neither the human nor venom PLA2's were directly injurious to target cells. However, in the presence of liposomal phospholipids, both PLA2's induced irreversible cell injury. Whereas venom PLA2 was cytolytic in the presence of either phosphatidylcholine or phosphatidylethanolamine (PE), rh-sPLA2 caused cell death only in the presence of PE. These data show that normal unperturbed cells are resistant to injury from PLA2, and that additional cofactors such as PE are required to induce cell injury. PMID- 9219903 TI - Effect of hyperoxia on the composition of the alveolar surfactant and the turnover of surfactant phospholipids, cholesterol, plasmalogens and vitamin E. AB - Experimental and clinical studies have provided evidence for the involvement of oxygen free radicals in development of acute and chronic lung diseases. Hyperoxia is very often an indispensable therapeutic intervention which seems to impose oxidative stress on lung tissue. We measured the effect of hyperoxia (80% O2 for 20 h) (1) on the lipid composition of pulmonary surfactant treated in vitro, (2) on surfactant lipid synthesis and secretion of type II pneumocytes in primary culture, (3) on the lipid composition and on the SP-A content of rat lung lavages and (4) on the turnover of phospholipids, cholesterol, plasmalogens and vitamin E in type II pneumocytes, lamellar bodies and lavages of adult rat lungs. (1) Hyperoxia of lung lavages in vitro reduces the vitamin E content significantly but does not change the relative proportion of PUFA or the content of plasmalogens. (2) Hyperoxia does not affect the biosynthesis or secretion of surfactant lipids and plasmalogens by type pneumocytes in primary culture. (3) Hyperoxic treatment of rats increases the SP-A content and reduces the vitamin E content significantly but does not change the concentration of other lipid components of lung lavage. (4) The vitamin E turnover, measured in type II pneumocytes, lamellar bodies and lung lavages, is increased 2-fold in these fractions. In contrast, the turnover of surfactant cholesterol and surfactant lipids does not change. (5) Hyperoxia caused an increase of the vitamin E uptake by type II pneumocytes resulting in a vitamin E enrichment of lamellar bodies. From these results we conclude that type II pneumocytes are able to regulate the turnover of lipophilic constituents of the alveolar surfactant independently of each other. Hyperoxia caused type II pneumocytes to increase the vitamin E content of lamellar bodies. The lipid and SP-A content of alveolar fluid can be regulated independently each other. PMID- 9219904 TI - Cloning and in vitro expression of rat lecithin:cholesterol acyltransferase. AB - Rat lecithin:cholesterol acyltransferase (LCAT) cDNA was obtained by reverse transcriptase/polymerase chain reaction amplification of rat liver total RNA. A consensus sequence was derived from four independent clones from two strains of rats. In vitro expression of rat LCAT cDNA in COS cells resulted in secreted enzyme protein with the same fatty acyl specificity for phospholipase A2 activity and cholesterol esterification as rat plasma LCAT, but different from that of recombinant or human plasma LCAT. PMID- 9219905 TI - Binding and uptake of chylomicron remnants by cultured arterial smooth muscle cells from normal and Watanabe-heritable-hyperlipidemic rabbits. AB - Chylomicron remnants (RM's) may be involved in atherogenesis because they can be delivered to the subendothelial space of arterial vessels and serve as substrate for arterial cells. A number of proteins may bind RM's, however, the quantitative significance of these is not established. The aim of this study was to identify the primary RM binding site of arterial smooth muscle cells (SMC's). At 4 degrees C, SMC's displayed saturable high affinity binding of RM's. In receptor competition studies, LDL inhibited binding of RM's by almost 60% suggesting involvement of the apolipoprotein B100/E receptor. Unlabeled RM's were more effective with an EC50 significantly less than for unlabeled LDL. Furthermore, at 37 degrees C RM uptake was three times greater than LDL, consistent with greater affinity of the apolipoprotein B100/E receptor for lipoproteins containing apolipoprotein E. In SMC's from homozygote Watanabe heritable hyperlipidemic (WHHL) rabbits, the binding and degradation of chylomicron remnants was severely impaired. SMC's from cross-bred WHHL rabbits exhibited levels of binding and degradation intermediate between homozygote WHHL rabbits and controls. We confirmed that the apolipoprotein B100/E receptor is the primary mechanism by which arterial smooth muscle cells bind and degrade RM's using a polyclonal antibody which specifically recognises the receptor. In the presence of the antibody, RM binding and degradation were inhibited by 90%. PMID- 9219906 TI - Studies to determine if rat liver contains multiple chain elongating enzymes. AB - According to the revised pathways of polyunsaturated fatty acid biosynthesis three, rather than two acids, must be chain elongated for converting linoleate and linolenate, respectively, to 22:5(n-6) and 22:6(n-3) (Sprecher et al. (1995) J. Lipid Res. 36, 2471-2477). The present study was undertaken to determine whether microsomes contained chain-length specific chain-elongating enzymes and, secondly, whether reaction rates for any of these reactions might be rate limiting in the synthesis of 24:5(n-6) and 24:6(n-3), which are the immediate precursors of 22:5(n-6) and 22:6(n-3). Rates of total chain elongation products produced from both 18:4(n-3) and 20:5(n-3) were about 3 nmol/min/mg of microsomal protein while only about 0.5 nmol/min/mg of 24:5(n-3) plus 24:6(n-3) was synthesized from 22:5(n-3). The rate of 24:5(n-3) synthesis was similar to that for the desaturation of 24:5(n-3), at position 6, to yield 24:6(n-3) (Geiger et al. (1993) Biochim. Biophys. Acta 1170, 137-142). The results suggest that the last chain elongation step in unsaturated fatty acid biosynthesis may be equally regulatory in governing the synthesis of fatty acids as is desaturation at position 6. When an enzyme saturating level of [1-(14)C]18:4(n-3) was incubated with increasing amounts of 18:3(n-6) there was a decrease in the production [1 (14)C]20:4(n-3). In a similar way it was observed that 18:4(n-3) inhibited the chain elongation of [1-(14)C]18:3(n-6). Identical cross-over inhibitory studies, using 20:4(n-6) and 20:5(n-3), as well as 22:4(n-6) and 22:5(n-3) also suggested that microsomes contain chain length specific chain-elongating enzymes. This conclusion was further supported by the finding that neither 20:5(n-3) or 22:5(n 3) inhibited the chain elongation of [1-(14)C]18:4(n-3). However, 18:4(n-3), and to a lesser degree, 22:5(n-3) did inhibit the chain elongation of [1-(14)C]20:5(n 3). This latter finding suggests that 18:4(n-3) and 20:5(n-3) might interact with the enzyme that chain elongates 20:5(n-3) to depress its ability to synthesize 22:5(n-3). Our results are most consistent with the presence of multiple chain elongating enzymes, but a more definitive answer requires the purification of these membrane-bound proteins. In addition our results suggest that the channeling of acids between enzymes in the endoplasmic reticulum may play an important role in regulating the biosynthesis of unsaturated fatty acids. PMID- 9219907 TI - Purification and partial sequencing of the XL-I form of xenobiotic-metabolizing medium chain fatty acid:CoA ligase from bovine liver mitochondria, and its homology with the essential hypertension protein. AB - The XL-I form of xenobiotic-metabolizing medium-chain fatty acid:CoA ligase was purified to apparent homogeneity from bovine liver mitochondria. The procedure gave rise to a 435-fold increase in specific activity, with a yield of 12%. The enzyme eluted from a gel filtration column as a single peak with an apparent molecular weight of ca. 55,000. It ran as a single band on SDS-polyacrylamide gel electrophoresis (SDS-PAGE) which had an apparent molecular weight of 62 kDa. N Terminal sequence analysis of the enzyme gave no sequence, which indicates a blocked N-terminus. To obtain sequence data, the enzyme was cleaved at methionine residues using CNBr. The resulting peptides were separated by SDS-PAGE. The cleavage pattern revealed two large peptides with molecular weights of ca. 10,000 and 12,000, plus several smaller peptides of lesser intensity. The 10 kDa and 12 kDa peptides were electroblotted onto Trans-Blot, and then sequenced directly from the blot. The N-terminal sequences of these two peptides are presented. When compared with known sequences it was discovered that these two peptides both have high homology with regions of the SA essential hypertension protein. This suggests a role for a carboxylic acid:CoA ligase in the control of high blood pressure. PMID- 9219908 TI - Effect of growth temperature on the biosynthesis of eukaryotic lipid molecular species by the cyanobacterium Spirulina platensis. AB - The incorporation of linoleic acid added at mmolar concentrations to the culture medium of the photosynthetic prokaryote Spirulina platensis results in the synthesis of membrane glycerolipids with a eukaryotic distribution of fatty acid chain length on the glycerol backbone (Pham Quoc et al., Biochim. Biophys. Acta [1993] 1168, 94-99). This distribution contrasts with the usual prokaryotic one found in lipids of cyanobacteria. A subsequent desaturation of the exogenously supplied fatty acid resulted in a large increase of gamma-linolenic acid. In order to estimate the capacities of S. platensis for bioconversion of fatty acids in lipid classes, the effects of different temperatures of growth were studied in linoleic acid-supplemented cultures. The lipid composition was affected by growth temperature, the synthesis of SQDG was stimulated at low temperature. The molecular species of each lipid were isolated and analyzed. Whatever the temperature of growth, the biosynthesis of eukaryotic C18/C18 lipid molecular species was observed in all lipid classes. Furthermore, the proportion of eukaryotic lipids increased at low temperature (24 degrees C). The desaturation of C18 fatty acids at C1 and C2 positions of the glycerol moiety occurred and was further stimulated when the growth temperature was lowered. The resulting proportion of gamma-linolenic acid increased significantly in cultures supplemented with linoleate at low temperatures. Finally a pathway for the synthesis of eukaryotic lipids and the desaturation of fatty acids esterified to the acyl lipids of linoleate-supplemented S. platensis can be suggested. PMID- 9219910 TI - Biosynthesis of furan fatty acids (F-acids) by a marine bacterium, Shewanella putrefaciens. AB - A mutant derived from Shewanella putrefaciens 8CS7-4 treated with N-methyl-N' nitro-N-nitrosoguanidine was found to produce 15-20 mg of a furan fatty acid (F acid), 10,13-epoxy-11-methyloctadeca-10,12-dienoic acid (F18), per liter of growth medium (10-15% of total fatty acids). Capillary gas chromatography-mass spectrometry and proton nuclear magnetic resonance analysis of the fatty acid methyl esters of the mutant revealed the presence of other F-acids, 8,11-epoxy-9 methylhexadeca-8,10-dienoic acid (F16), 6,9-epoxy-7-methyltetradeca-6,8-dienoic acid (F14), and methyl branched unsaturated fatty acids, 11-methyl-12E octadecenoic acid (11-me-18:1) and 11-methyl-10E,12E-octadecadienoic acid (1-me 18:2). About 90% of F-acids were present in phospholipids, in which the F-acids were found to be exclusively linked at the sn-1 position. 11-me-18:1 and 11-me 18:2 were also detected in the sn-1 position. Firstly, 11-me-18:1 increased and reached a maximum at 12 h, and then decreased rapidly. Secondly, the 11-me-18:2 content reached a maximum at 24 h, when 11-me-18:1 was little detected, and then decreased. Finally, the amount of F18 began to increase after 20 h and reached a plateau at 36 h. These results suggest that 11-me-18:1 and 11-me-18:2 are precursors of F18. PMID- 9219909 TI - Omega-3 fatty acid intake results in a relationship between the fatty acid composition of LDL cholesterol ester and LDL cholesterol content in humans. AB - The relationship between the fatty acid composition of the low density lipoprotein (LDL) cholesterol ester and LDL cholesterol content was assessed in 26 free-living, normal subjects. Dietary intakes of 14:0, 16:0, 18:0, 18:1, 18:2omega6, 18:3omega3, 20:4omega6, 20:5omega3, 22:6omega3 were calculated from seven-day food records kept by each subject at baseline and after three months of supplementation with olive, flaxseed or fish oil, respectively. A randomized cross-over design was used. The fatty acid content of specific foods was calculated. Fasting blood samples, taken at the beginning and end of each supplementation period, were analyzed for the fatty acid content present in individual lipoproteins. There was a significant correlation between 20:5omega3 and 22:6omega3 intake and the content of these fatty acids in the LDL cholesterol ester fraction. During the fish oil treatment period the 16:0 and 18:0 content of the LDL cholesterol ester was highly predictive of LDL cholesterol content. This relationship was not observed during the baseline or placebo (olive oil) supplement period. PMID- 9219911 TI - Association of Lp(a) rather than integrally-bound apo(a) with triglyceride-rich lipoproteins of human subjects. AB - The majority of apolipoprotein (a) [apo(a)] in plasma is characteristically associated with Lipoprotein (a) [Lp(a)], having a buoyant density (1.05-1.08 g/ml) intermediate between low density lipoproteins (LDL) and high density lipoproteins (HDL). In the fed (postprandial) state or in the presence of fasting (endogenous) hypertriglyceridemia, a small proportion of plasma apo(a) is found in the density < 1.006 g/ml fraction of plasma, associated with larger and less dense triglyceride-rich lipoproteins (TRL). In order to further characterize the presence of apo(a) in ultracentrifugally-separated TRL (UTC-TRL), this lipoprotein fraction was isolated from plasma obtained in the fed state (three hours after an oral fat load) from healthy normolipidemic subjects (Lp(a): 38 +/- 8 mg/dl (mean +/- S.E.), n = 4) and also from plasma obtained after an overnight fast from hypertriglyceridemic patients (plasma TG: 8.16 +/- 2.00 mmol/l, Lp(a): 41 +/- 3 mg/dl, n = 18). Apo(a) in 3 h-postprandial UTC-TRL (5 +/- 2% of total plasma apo(a)) and in hypertriglyceridemic UTC-TRL (8 +/- 2% total apo(a)) was separable by electrophoresis and/or gel chromatography (FPLC) from the majority of UTC-TRL lipid. Apo(a) in UTC-TRL fractions had slow pre-beta electrophoretic mobility and was isolated in a lipoprotein size-range smaller than VLDL and larger than LDL, consistent with it being Lp(a). Recentrifugation of UTC-TRL resulted in the majority of apo(a) being recovered in the density > 1.006 g/ml fraction. Addition of proline to plasma samples before ultracentrifugation (final concentration: 0.1 M) substantially reduced the amount of Lp(a) in UTC-TRL. TRL separated from plasma by FPLC contained less apo(a) (2-5% of total plasma apo(a)), but this apo(a) was also readily dissociable from TRL lipid, had slow pre-beta electrophoretic mobility, and was associated with a lipoprotein with the size of Lp(a). Our data suggest that apo(a) in the TRL fraction of subjects with postprandial triglyceridemia or endogenous hypertriglyceridemia is not an integral component of plasma VLDL or chylomicrons, but represents the presence of non-covalently bound Lp(a). PMID- 9219912 TI - Mycolic acid biosynthesis: definition and targeting of the Claisen condensation step. AB - Through the use of 2,2-[2H]palmitic acid pulse labeling of the whole cells of C. matruchotti and analysis by gas chromatography-mass spectrometry of the non labeled and [2H]-labeled corynomycolates, we established a new mechanism for palmitate condensation devoid of the postulated carboxylation step. This evidence allowed the design and synthesis of several structurally related antagonists against the condensation reactions which were shown to possess potent in vivo activity against C. matruchotti with complete inhibition of growth on solid media at concentrations between 1-10 microg/ml. In addition, a cell-free in vitro assay of corynomycolate synthesis was developed to allow the screening of these and other antagonists. PMID- 9219913 TI - Limited proteolysis of high density lipoprotein abolishes its interaction with cell-surface binding sites that promote cholesterol efflux. AB - High-density lipoprotein (HDL) components remove cholesterol from cells by two independent mechanisms. Whereas HDL phospholipids pick up cholesterol that desorbs from the plasma membranes, HDL apolipoproteins appear to interact with cell-surface binding sites that target for removal pools of cellular cholesterol that feed into the cholesteryl ester cycle. Here we show that mild trypsin treatment of HDL almost completely abolishes this apolipoprotein-mediated cholesterol removal process. When HDL was treated with trypsin for various periods of time and then incubated with cholesterol-loaded fibroblasts, treatment for only 5 min reduced the ability of HDL to remove excess cholesterol from cellular pools that were accessible to esterification by the enzyme acyl CoA:cholesterol acyltransferase. This mild treatment digested less than 20% of HDL apolipoproteins and did not alter the lipid composition, size distribution, or electrophoretic mobility of the particles. Trypsin treatment of HDL for up to 1 h caused no further reduction in its ability to remove cellular cholesterol despite a greater than 2-fold increase in apolipoprotein digestion. Trypsin treatment of HDL also reduced its ability to deplete the cholesteryl ester content of sterol-laden macrophages. Promotion of cholesterol efflux from the plasma membrane by HDL phospholipids was unaffected by even extensive proteolysis. In parallel to the loss of cholesterol transport-stimulating activity, trypsin treatment of HDL for only 5 min nearly abolished its interaction with high-affinity binding sites on cholesterol-loaded fibroblasts. Reconstitution of trypsin-modified HDL with isolated apo A-I or apo A-II restored the cholesterol transport-stimulating activity of the particles. Thus a minor trypsin-labile fraction of HDL apolipoproteins is almost exclusively responsible for the apolipoprotein-dependent component of cholesterol efflux mediated by HDL particles. PMID- 9219915 TI - Docosahexaenoic acid causes accumulation of free arachidonic acid in rat pineal gland and hippocampus to form hepoxilins from both substrates. AB - Hepoxilins (Hx) are biologically active metabolites of arachidonic acid (AA) formed regioselectively from 12(S)-HPETE by 'hepoxilin synthase'. Hx modulate synaptic neurotransmission in hippocampal CA1 neurons, and inhibit norepinephrine release in hippocampal slices. During the course of our studies we investigated whether docosahexaenoic acid (DHA) was a substrate for hepoxilin formation. We used two tissues, the pineal gland and hippocampal slices. Tissues were incubated alone or with AA (20 microg/ml) or DHA (20 microg/ml). After 60 min at 37 degrees C, samples were acid-extracted to convert Hx into their stable trioxilin (TrX) form and analyzed as the Me-TMSi derivatives by EI-GC/MS to determine the structures of the DHA metabolites, and as PFB-TMSi derivatives by GC/MS in the NICI mode using SIM to simultaneously quantify TrX products of the 3-series (derived from AA) monitored at m/z 569, while those of the 5-series (derived from DHA) were monitored at m/z 593. Results show good conversion of both substrate fatty acids by the rat pineal gland and hippocampal slices, into the 3-series (21.3 +/- 5.8 and 12.5 +/- 2.2 ng/microg protein, respectively) and 5-series TrX (12.3 +/- 2.7 and 2.9 +/- 0.4 ng/microg protein, respectively). Surprisingly though, experiments with DHA, in both tissues, also showed formation of TrX derived from endogenous AA (3-series) (10.4 +/- 8.3 and 3.1 +/- 2.1 ng/microg protein, respectively). These experiments demonstrate previously unreported actions of DHA causing the accumulation of AA, which is converted into hepoxilins. In order to prove that AA is accumulated during DHA stimulation of the tissue, we carried out separate experiments with hippocampal slices in which the neutral lipids and phospholipids were labeled with [14C]AA. DHA caused a time dependent appearance of free [14C]AA which was released mostly from the TG pool. Measurement of the AA/DHA ratio in the TG pool by GC/MS further indicated that DHA is incorporated into the TG at the expense of AA. These results demonstrate that DHA competes with AA for acylation into the metabolically active TG fraction, and both fatty acids are converted into hepoxilins of the corresponding series. PMID- 9219914 TI - Tyrosine phosphorylation of 100-115 kDa proteins by phosphatidic acid generated via phospholipase D activation in HL60 granulocytes. AB - In HL60 granulocytes, 4beta-phorbol 12-myristate 13-acetate (PMA) induced tyrosine phosphorylation of several proteins with molecular weight of 100-115 kDa and 45 kDa. Furthermore, PMA-mediated phosphatidic acid (PA) production via phospholipase D (PLD) activation. In the presence of either butanol or ethanol, PMA-induced PA production was markedly reduced and instead a metabolically stable phosphatidylbutanol (PBut) or phosphatidylethanol (PEt) was produced by transphosphatidylation by PLD. Under the same incubation condition, these primary alcohols inhibited PMA-induced tyrosine phosphorylation of the 100-115 kDa proteins. Propranolol, which is often used as a selective inhibitor of PA phosphohydrolase (PAP) involving diacylglycerol (DG) formation from PA, did not affect tyrosine phosphorylation of the 100-115 kDa proteins. Moreover, incubation of HL60 granulocytes with Streptomyces chromofuscus PLD caused both PA production and tyrosine phosphorylation of the above proteins. Exogenous PA treatment also induced tyrosine phosphorylation of the same proteins. Thus, the results presented here suggest that PA produced via PLD activation is involved in tyrosine phosphorylation of the 100-115 kDa proteins in HL60 granulocytes. PMID- 9219916 TI - Estrogenic activity is increased for an antiestrogen by a natural mutation of the estrogen receptor. AB - The estrogen receptor (ER) functions as a ligand-activated transcription factor which mediates the actions of estrogens and antiestrogens in target tissues. Other investigators have shown that artificial point mutations in the transcriptional activation domain AF-2 of the ligand binding domain (LBD) of the ER can increase the estrogenic properties of antiestrogens, determined by transcriptional activation of estrogen-responsive reporter constructs cotransfected into cells. Although these data provide valuable information about ER function there is no evidence that these mutations occur naturally. We have taken a different approach and examined the naturally occurring codon 351 asp --> tyr mutation in the LBD of ER to stimulate the expression of an endogenous target gene. This approach avoids dependence on artificial reporter constructs and their idealized estrogen response elements (EREs). In this report we describe the regulation of transforming growth factor alpha (TGF alpha) mRNA by estradiol and the antiestrogens keoxifene and ICI 182,780 in our stable transfectants of ER negative MDA-MB-231 breast cancer cells, which express either the wild-type (S30 cells) or codon 351 asp --> tyr mutant ER (BC-2 cells). The mutant receptor was identified in a tamoxifen-stimulated human breast tumor. Our results demonstrate, for the first time, that a naturally occurring mutation in the ER changes the pharmacology of the antiestrogen keoxifene by increasing estrogenic activity, and that keoxifene exhibits a gene-specific estrogen-like effect with mutant ER but not with wild-type ER. The pure antiestrogen ICI 182,780 maintained complete antagonistic activities in both ER transfectants, demonstrating that its action is unaffected by the mutation. PMID- 9219917 TI - Interaction between estradiol and growth factors in the regulation of specific gene expression in MCF-7 human breast cancer cells. AB - The response of two endogenous, estrogen-induced genes, LIV-1 and pS2, to growth factor stimulation of MCF-7 cells was examined. Epidermal growth factor (EGF), transforming growth factor alpha (TGF alpha) and insulin-like growth factor-1 (IGF-1) were each able to induce an increase in the two mRNAs in the absence of estradiol, and their effects were additive to that of an optimally inducing concentration (10(-8) M) of the hormone. Induction by EGF and TGF alpha, but not by IGF-1, were also additive to induction by a saturating concentration (2 microg/ml) of insulin. TGFbeta, an antimitogenic growth factor for MCF-7 cells, did not induce LIV-1 or pS2 mRNA but inhibited induction by estradiol. Increases in mRNA were shown to reflect increases in specific gene transcription. Induction by growth factors, but not by estradiol, was dependent upon protein synthesis. Induction by both growth factors and estradiol was inhibited by the pure antiestrogen, ICI 164384 (ICI), and by the mixed agonist/antagonist, tamoxifen. Despite differences in patterns of expression in vivo and in vitro, both LIV-1 and pS2 appeared to be responsive to growth factors via a mechanism distinct from that of estradiol but requiring the estrogen receptor. PMID- 9219918 TI - Lack of relationship between the expression of Hsp27 heat shock estrogen receptor associated protein and estrogen receptor or progesterone receptor status in male breast carcinoma. AB - Estrogen, through estrogen receptors (ERs), may regulate the synthesis of progesterone receptors (PRs) and of a heat shock estrogen receptor-associated protein (hsp27). In female breast carcinoma (FBC) both proteins serve as surrogate indicators for the presence of functional ERs. In addition, the expression of these proteins was related to other prognostic indicators of value in female breast tumours. Endocrine disorders, hormone therapy and altered estrogen metabolism have been associated with the development of male breast cancer (MBC), suggesting that evaluation of the expression of ER, PR and hsp27 might improve our understanding of the biology of this tumour. ER and PR status and hsp27 expression were evaluated by immunohistochemistry in 16 primary MBC patients. The interrelationships between these parameters were established and compared with the clinicopathological data on the tumours. Ten (56%) MBC patients were ER-positive, 69% were PR-positive and all samples were hsp27-positive. Our series of MBC patients showed a positive correlation between ERs and PRs, however there was a lack of correlation between hsp27 and ERs or PRs. MBCs did not exhibit any correlation between the biomarkers studied and known prognostic indicators for females (e.g. Scarff-Bloom-Richardson (SBR) or modified SBR (MSBR) grade, T stage, lymph node status). This is the first published series reporting the incidence of hsp27 in MBC. The lack of association between the expression of ERs and hsp27 found in MBC differs from the results reported for FBC, moreover the expression of ERs, PRs or hsp27 did not correlate with the clinicopathological parameters that have prognostic value in females. Although the data were obtained from a relatively small sample population, our findings suggest that MBC and FBC are biologically different tumours with respect to the expression of the studied proteins. PMID- 9219919 TI - Analysis of ligand dependence and hormone response element synergy in transcription by estrogen receptor. AB - In this work we examined two questions: (1) Is the low, but readily detectable, ability of estrogen receptor (ER) to activate transcription in the absence of added 17beta-estradiol caused by traces of estrogen in the growth medium, or by a weak ligand-independent ability of ER to activate transcription? (2) Does the ER exhibit synergistic activation of transcription on reporter genes containing multiple estrogen response elements (EREs)? To study these questions we developed a powerful new reporter gene, containing four EREs, which achieves inductions of up to 330-fold in the presence of liganded ER. We provided several types of evidence indicating that under standard cell culture conditions unliganded ER is unable to activate transcription. We demonstrated that when cells are grown in serum-free medium, estrogenic compounds may be in the base tissue culture medium. We demonstrated a strong cell and ER-dependence in transcriptional synergy, and suggest that cooperative binding of ER to multiple EREs can be responsible for transcriptional synergy in vivo. PMID- 9219920 TI - Gonadal sex differentiation in chicken embryos: expression of estrogen receptor and aromatase genes. AB - Estrogen is implicated in sexual differentiation of the avian gonad. Expression of the estrogen receptor and aromatase genes was therefore examined at the time of gonadal sex differentiation in chicken embryos, using reverse transcription and the polymerase chain reaction (RT-PCR). Estrogen receptor (cER) transcripts were detected in the gonads of both presumptive sexes at embryonic days 4.5, 5.5 and 6.5, and in female but not male urogenital tissues at day 3.5. Aromatase (cAROM) transcripts were detected in female but not male gonads from day 6.5 of embryogenesis, and in adult gonads of both sexes. Both female and male embryos thus express cER mRNA before morphological differentiation of the gonads, which begins on day 5, whereas cAROM expression begins at or shortly after the onset of differentiation and is female-specific. Examination of other tissues showed that, in 5.5-day-old embryos, cER expression was limited to the gonads; no transcripts were detected in the mesonephric kidney, liver, brain, hindlimb or heart of either sex. In 9.5-day-old female embryos, cER and cAROM transcripts were present in both the left (ovarian) and the right (regressing) gonads. Altogether, these observations imply that the gonads of both sexes develop the capacity to respond to estrogens early in embryogenesis, before morphological differentiation, whereas the capacity to synthesize estrogens is female-specific and occurs later, at the time of differentiation. These observations are consistent with estrogens having a key role in ovarian development. PMID- 9219921 TI - 4-Methyl-3-oxo-4-aza-5alpha-androst-1-ene-17beta-N-aryl-carboxamides: an approach to combined androgen blockade [5alpha-reductase inhibition with androgen receptor binding in vitro]. AB - 4-Aza-5alpha-androstan-3-one 17beta-(N-substituted carboxamides) are potent human type 2 5alpha-reductase (5aR) inhibitors with generally poor binding to the human androgen receptor (hAR). When the 17-amide N-substituent included an aromatic residue, potent dual inhibitors of both type 1 and 2 5aR are produced, but hAR binding remained poor. Tertiary-substituted-17-amides have reduced inhibition of both 5aR isozymes. The addition of an N4-methyl substitutent to the A-ring profoundly increased hAR affinity and the addition of unsaturation to the A-ring (delta1) modestly augmented hAR binding. The unsubstituted carbanilides in the delta1-N4-methyl series show some selectivity for type 1 5aR over the type 2 isozyme, whereas addition of aryl substituents, particularly at the 2-position, increased type 2 5aR binding to provide dual inhibitors with excellent hAR binding, e.g. N-(2-chlorophenyl)-3-oxo-4-methyl-4-aza-5alpha-androst-1-ene-17bet a-carboxamide (9c). Compounds of this type exhibit low nanomolar IC50s for both human 5aR isozymes as well as the human androgen receptor. Kinetic analysis confirms that the prototype 9c displays reversible, competitive inhibition of both human isozymes of 5aR with K(i) values of less than 10 nM. Furthermore, this compound binds to the androgen receptor with an IC50 equal to 8 nM. Compounds in this series are projected to be powerful antagonists of testosterone and dihydrotestosterone action in vivo, with potential utility in the treatment of prostatic carcinoma (PC). PMID- 9219922 TI - Regional distribution of cytosolic and particulate 5alpha-dihydroprogesterone 3alpha-hydroxysteroid oxidoreductases in female rat brain. AB - Numerous studies have indicated that progesterone metabolites, particularly 3alpha,5alpha-tetrahydroprogesterone, can potently influence multiple brain functions, e.g. they have the capacity to mediate gonadotropin regulation and various anticonvulsive, anesthetic and anxiolytic effects. These circulating progesterone metabolites are likely to represent only a fraction of the bioavailable pool of these steroids in that the central nervous system (CNS) also possesses enzymes that can synthesize these metabolites in situ. Therefore, because the ability of the CNS to produce these neuroactive progestins is an important consideration when assessing overall progestin function and metabolism, we measured the major progesterone metabolizing enzyme activities, namely the cytosolic NADPH and particulate NADH 5alpha-dihydroprogesterone 3alpha hydroxysteroid oxidoreductase (3alpha-HSOR) and progesterone 5alpha-reductase activities in nine brain regions from random cycling and ovariectomized rats. These assays entailed the use of reverse isotopic dilution analysis and revealed that all three enzymic activities were present in each of the brain regions examined, but that these regions displayed differential patterns with regard to their levels of cytosolic and particulate 3alpha-HSOR activity. The cytosolic 3alpha-HSOR activity was highest in the olfactory bulb/tubercle and colliculi regions which were greater than levels in the hypothalamus/preoptic area and cerebellum which were greater than levels in the amygdala/striatum and hippocampus/dentate gyrus. Midbrain/thalamus, cerebral cortex and pons/medulla were different only from the olfactory bulb/tubercle and colliculi regions. The particulate 3alpha-HSOR activity was highest in the olfactory bulb/tubercle region followed by colliculi, hippocampus/dentate gyrus and pons/medulla which were greater than levels in the hypothalamus/preoptic area, cerebellum and amygdala/striatum. Cerebral cortex and midbrain/thalamus were different only from the olfactory bulb/tubercle area. The highest levels of 5alpha-reductase activity were found in the pons/medulla region followed by the colliculi, midbrain/thalamus, cerebellum and olfactory bulb/tubercle which were greater than levels in the amygdala/striatum, hippocampus/dentate gyrus, hypothalamus/preoptic area and cerebral cortex. It is interesting to note that although 5alpha reductase may control, at least in part, substrate levels for the 3alpha-HSORs, the distribution of 5alpha-reductase activity in these nine brain regions did not correlate with 3alpha-HSOR levels. The differences in the levels of activity of these three enzymes in various brain regions suggests a role in maintaining a differential balance of the neuroactive steroid, 3alpha,5alpha tetrahydroprogesterone, and its precursor, 5alpha-dihydroprogesterone, in various regions of the CNS. PMID- 9219923 TI - Long-term corticosteroid treatment but not chronic stress affects 11beta hydroxysteroid dehydrogenase type I activity in rat brain and peripheral tissues. AB - Long-term treatment (21 days) of male rats with corticosterone in the drinking water caused a significant increase in the activity of the NADP-dependent form of 11beta-hydroxysteroid dehydrogenase (11-HSD1) in the pituitary, thymus, and spleen, (marginally in the hippocampus, amygdala and lymph nodes), without having any effect in a number of other central and peripheral tissues. In contrast, repeated restraint stress, although increasing plasma corticosterone to the same level as that observed after its administration, failed to change the activity of this key regulatory enzyme, which allows aldosterone to exert its specific effects in the presence of a large excess of corticosterone. This resistance to elevation in 11-HSD activity was also observed in the thymuses of subordinate rats during social stratification in a visible burrow system. In both cases, the circulating levels of corticosterone were much higher in stressed rats than in control animals. Factors which might account for these differences in response are discussed and compared with the situation in intact cells where, unlike in tissue homogenates, the reduction of 11-dehydrocorticosterone to corticosterone (reductase activity) appears to predominate. PMID- 9219924 TI - Expression and distribution of cortical type aromatase mRNA variant in the adult rat brain. AB - Our previous study showed that a relatively high level of the aromatase mRNA existed in the cerebral cortex (CC) of the rat, where the aromatase activity was reported to be little or absent. To elucidate the identity of the aromatase mRNA in the CC of the rat, we investigated the 5'-region of the aromatase mRNA in the rat CC. When the sequence of the 5'-region of the cortical message was analysed by the 5'-rapid amplification of cDNA ends (5'-RACE) with the antisense primer for exon II using the RNA extracted from the CC, no clone could be isolated. However, the upstream sequence from the 5'-end of exon IV of the aromatase clones, isolated from the CC by RACE with the antisense primers for exon V, was different from that on the aromatase mRNA encoding the full translated region. The new sequence of the cortical type message, called the cortical type aromatase mRNA variant, was located on the intron upstream of exon IV in the genomic cDNA. Distribution of the brain aromatase message with exons III-V and the cortical type aromatase mRNA variant were analysed by the reverse transcription-polymerase chain reaction (RT-PCR) using total RNAs extracted from the hypothalamus-preoptic area (HPOA), amygdala (AMY) and CC. The PCR products with primers for exons III-V were generated from the HPOA and AMY, but not from the CC. On the other hand, the PCR products with primers for exon IIIv (cortical type aromatase mRNA variant specific)-V were detected in significant amounts in the CC as well as the HPOA and AMY. These results indicate the existence of the aromatase mRNA variant lacking exons I-III in the adult rat brain. This cortical type mRNA variant seemed to be widely distributed in the tissues. PMID- 9219925 TI - Origin of deoxycorticosterone sulfate (DOC-SO4) in plasma of pregnant women: pregnenolone-3,21-disulfate is a placental precursor of DOC-SO4. AB - The plasma levels of deoxycorticosterone sulfate (DOC-SO4) in near-term pregnant women are approximately 100 times those in plasma of men or non-pregnant women. Yet, neither the tissue site of synthesis nor the precursor of DOC-SO4 that enters maternal plasma is known. Several potential sources have been excluded: plasma DOC-SO4 is not derived from plasma DOC; and the secretion of C21-steroids (other than aldosterone) from the maternal adrenals during human pregnancy is not increased. Similarly, the transfer of DOC-SO4 from fetal plasma cannot account for the high level of DOC-SO4 in the maternal compartment, and a reduced clearance of plasma DOC-SO4 during pregnancy cannot account for the high levels of DOC-SO4. Indeed, the rate of clearance of DOC-SO4 from plasma is 10-100 times that of most other steroid sulfates. To address this question further, we evaluated the possibility that fetal plasma pregnenolone-3,21-disulfate serves as a precursor for DOC-SO4 formation in the placenta. The preferential hydrolysis of the 3beta-sulfate of pregnenolone-3,21-disulfate in placenta would give rise to pregnenolone-21-monosulfate, which, if acted upon by placental 3beta hydroxysteroid dehydrogenase/delta5 --> 4 isomerase, could give DOC-SO4. [3H]Pregnenolone-3,21-disulfate was incubated with minces of human placental tissue for 5, 20, 60 and 120 min. Radiolabelled DOC-SO4, DOC, and pregnenolone-21 monosulfate were isolated from the incubation media and quantified. After a 5 min incubation, 7.5% of substrate was converted to DOC-SO4; and after 20, 60 and 120 min approximately 30% of the [3H]pregnenolone-3,21-disulfate was recovered from the media of these incubations as [3H]DOC-SO4. [3H]DOC was also present in the incubation media and the concentrations of this product increased as a function of incubation time. Therefore, pregnenolone-3,21-disulfate, which is present in very high concentrations in fetal plasma (approximately 1000 ng/ml), is metabolized in the placenta to DOC-SO4. Because of the fetal and maternal vascular arrangements of the hemochorioendothelial placenta of human pregnancy, steroids produced in syncytiotrophoblasts preferentially enter the intervillous space; thus, fetal plasma pregnenolone-3,21-disulfate may serve as a placental precursor of maternal plasma DOC-SO4. PMID- 9219926 TI - Involvement of cytochrome P-450 in the 15alpha-hydroxylation of 13-ethyl-gon-4 ene-3,17-dione by Penicillium raistrickii. AB - The 15alpha-hydroxylation of 13-ethyl-gon-4-ene-3,17-dione (GD) with different subcellular fractions of Penicillium raistrickii i 477 was investigated. Cytochrome P-450 was shown to be involved in this reaction. The steroid transformation was inhibited by carbon monoxide, metyrapone, p-CMB, iodoacetamide, N-methylmaleimide and several metal ions. The 15alpha-hydroxylase was observed to be dependent on nicotinamide-adenine dinucleotide phosphate (NADPH) replaceable by NaIO4, and the activity was enhanced by a NADPH regenerating system, indicating the involvement of the NADPH-cytochrome c (P-450) reductase. This was further confirmed by the inhibition of the hydroxylase activity in the presence of cytochrome c. No effect was observed in the presence of azide and antimycin A. Solubilized microsomes gave an absorption maximum at 453 nm in carbon monoxide difference spectrum, and showed a Type-I GD-binding spectrum typically for cytochrome P-450 interaction with substrate. First results about the inducibility of the enzymes involved in the 15alpha-hydroxylation of GD are shown. PMID- 9219927 TI - Biochemistry and pharmacokinetics of potent non-steroidal cytochrome P450(17alpha) inhibitors. AB - Two potent non-steroidal inhibitors (CB7645 and CB7661) of human cytochrome P450(17alpha) were tested for in vivo activity in WHT mice. There were no signs of toxicity, but there was no effect on the androgen-dependent organs. The pharmacokinetics and biochemistry of the compounds in mice were investigated. Following i.p. administration of 0.5 mmol/kg of CB7645 and CB7661, peak plasma levels of 13.4 and 3.4 microM, respectively, occurred after 2-4 h, both compounds were cleared rapidly (terminal half-lives 2.7 and 3.3 h, respectively) and neither was detectable at 24 h. CB7645 produced some decrease in plasma testosterone at 4 h, but this was not sustained. When tested in vitro against the WHT testicular enzyme, the CB7645 and CB7661 were competitive inhibitors with K(i) values of 10 and 13 nM, respectively. However, the K(m) for the substrate progesterone was lower at 4.3 nM. These data indicate that, for effective and continuous inhibition of the murine cytochrome P450(17alpha) enzyme, higher peak levels of the compounds would be required, and these levels would need to be maintained throughout the treatment period. PMID- 9219928 TI - The effects of progesterone, 4,16-androstadien-3-one and MK-434 on the kinetics of pig testis microsomal testosterone-4-ene-5alpha-reductase activity. AB - The enzyme 3-oxo-steroid: NADP+ 4-oxidoreductase (EC 1.3.1.22; 5alpha-reductase) was assayed in testicular microsomes of pigs of 3, 20 and 24 weeks of age. The activity was very low in 3-week-old animals and approximately 10-fold higher in 5 and 6-month-old pigs. The pH optimum was 6.3 in 6-month-old animals, 5.7 in 5 month-old animals, but could not be reliably determined in 3-week-old animals. The kinetic parameters for 5alpha-reductase in testis microsomes from 6-month-old animals were; K((m)(app)), 8.0 micromol/l, V((max)(app)), 6.7 nmoles/90 min/mg protein. Progesterone was a competitive inhibitor of testosterone 5alpha reduction with an apparent K((i)(app)) of 0.86 micromol/l. However, 4,16 androstadien-3-one (dienone), which undergoes 5alpha-reduction in the biosynthesis of the pheromonally active 16-androstenes, was a comparatively poor inhibitor with a K((i)(app)) of 4.9 micromol/l. Similarly, MK434, which is a selective inhibitor of the human type 2 5alpha-reductase, but which inhibits both types 1 and 2 in the rat, was also a poor competitive inhibitor of testosterone 5alpha-reductase in the pig testis (K((i)(app)), 3.1 micromol/l). It would appear from these studies that the pig testis microsomal 5alpha-reductase corresponds to a type 1 isozyme that is not capable of reducing dienone other than under conditions where the dienone concentration would be in considerable excess of testosterone. It is, therefore, probable that substrate-specific 5alpha reductases exist in the pig testis for the 5alpha-reduction of testosterone and dienone. PMID- 9219929 TI - Synaptic effects of identified interneurons innervating both interneurons and pyramidal cells in the rat hippocampus. AB - GABAergic interneurons sculpt the activity of principal cells and are themselves governed by GABAergic inputs. To determine directly some of the sources and mechanisms of this GABAergic innervation, we have used dual intracellular recordings with biocytin-filled microelectrodes and investigated synaptic interactions between pairs of interneurons in area CA1 of the adult rat hippocampus. Of four synaptically-coupled interneuron-to-interneuron cell pairs, three presynaptic cells were identified as basket cells, preferentially innervating somata and proximal dendrites of pyramidal cells, but one differing from the other two in the laminar distribution of its dendritic and axonal fields. The fourth presynaptic interneuron was located at the border between strata lacunosum moleculare and radiatum, with axon ramifying within stratum radiatum. Action potentials evoked in all four presynaptic interneurons were found to elicit fast hyperpolarizing inhibitory postsynaptic potentials (mean amplitude 0.35 +/- 0.10 mV at a membrane potential of -59 +/- 2.8 mV) in other simultaneously recorded interneurons (n=4). In addition, three of the presynaptic interneurons were also shown to produce similar postsynaptic responses in subsequently recorded pyramidal cells (n=4). Electron microscopic evaluation revealed one of the presynaptic basket cells to form 12 synaptic junctions with the perisomatic domain (seven somatic synapses and five synapses onto proximal dendritic shafts) of the postsynaptic interneuron in addition to innervating the same compartments of randomly-selected local pyramidal cells (50% somatic and 50% proximal dendritic synapses, n=12). In addition, light microscopic analysis also indicated autaptic self-innervation in basket (12 of 12) and bistratified cells (six of six). Electron microscopic investigation of one basket cell confirmed six autaptic junctions made by five of its boutons. Together, these data demonstrate that several distinct types of interneuron have divergent output to both principal cells and local interneurons of the same (basket cells) or different type. The fast synaptic effects, probably mediated by GABA in both postsynaptic interneurons and principal cells are similar. These additional sources of GABA identified here in the input to GABAergic cells could contribute to the differential temporal patterning of distinct GABAergic synaptic networks. PMID- 9219930 TI - L-arginine potentiates GABA-mediated synaptic transmission by a nitric oxide independent mechanism in rat dopamine neurons. AB - Effects of L-arginine in the nervous system are often attributed to nitric oxide. Using whole-cell patch pipettes to record membrane currents in voltage-clamp from dopamine neurons in the rat midbrain slice, the present studies found that L arginine potentiates GABA-dependent membrane currents via a nitric oxide independent mechanism. L-Arginine (0.3-10 mM) increased the peak amplitude, half width duration and time constant of decay of GABA(B) receptor-mediated inhibitory postsynaptic currents in a concentration-dependent manner. In the presence of CGP 35348 (300 microM), a GABA(B) receptor antagonist, L-arginine also prolonged the duration of inhibitory postsynaptic currents mediated by GABA(A) receptors, but their amplitudes were reduced. L-Arginine (10 mM) also evoked 17+/-3 pA of outward current (at -60 mV) which was significantly increased in the presence of exogenous GABA (100 microM). Pressure-ejection of GABA from micropipettes produced outward currents mediated by GABA(B) receptors (recorded in bicuculline) or GABA(A) receptors (recorded in CGP 35348); both types of receptor-mediated currents were increased by L-arginine (10 mM). In contrast, outward currents evoked by baclofen, a GABA(B) receptor agonist, were not potentiated by L arginine. The GABA transport inhibitors NO 711 (1 microM) and nipecotic acid (1 mM) significantly increased the half-width duration and time-constant of decay of GABA(B)-mediated inhibitory postsynaptic currents, thus mimicking effects of L arginine. However, nitric oxide donors failed to mimic effects of L-arginine on GABA(B) inhibitory postsynaptic currents, and inhibitors of nitric oxide synthesis failed to selectively block the action of L-arginine. These findings suggest that L-arginine potentiates GABA synaptic transmission by a nitric oxide independent mechanism. Similarities between effects of L-arginine, NO 711 and nipecotic acid suggest that L-arginine inhibits a GABA transporter. PMID- 9219931 TI - Interactions between nitric oxide and dopamine in inhibitory learning and memory in newborn rats. AB - Taking into account our previous results on dopamine and nitric oxide effects on neonatal inhibitory learning and memory in rats, the mutual interactions of the two molecules were studied in this experimental paradigm. Both increased dopamine content and nitric oxide bioavailability in the brain after application of dopamine and L-arginine as substrate for nitric oxide synthase solutions into lateral cerebral ventricles improved learning and 24 h memory. Joint application of dopamine and L-arginine yielded still more improvement. Learning and memory processing were dose dependently enhanced by D1 receptor agonists as well, whereas D1 receptor antagonists had an opposite and also dose-dependent effect. Dopamine or D1 receptor agonists administered together with nitro-L-arginine, a nitric oxide synthase inhibitor that impaired learning and memory due to a decreased nitric oxide availability, antagonized the effect of nitro-L-arginine, as did L-arginine. D1 receptor antagonists impaired both learning and memory, and L-arginine rendered learning values normal. The dopamine and D1 receptor-agonist effect on 24 h memory was concentration dependent, and their higher concentrations substantially increased the retention indexes. The intimate mechanisms of these interactions are to be identified in further experiments. PMID- 9219932 TI - Nicotinic receptor mediates spontaneous GABA release in the rat dorsal motor nucleus of the vagus. AB - Spontaneous postsynaptic currents were investigated in neurons of the caudal portion of the dorsal motor nucleus of the vagus using the patch-clamp technique to study the effect of neuronal nicotinic acetylcholine receptor activation on synaptic transmission. In voltage-clamped neurons, bath application of nicotine (1-30 microM) elicited a concentration-dependent increase in the frequency of the spontaneous synaptic currents. The effect was also observed with application of the nicotinic receptor agonists epibatidine (10 nM) and cytisine (10 microM). Mecamylamine (20 microM) and curare (50 microM), two nicotinic receptor antagonists, both decreased the effect of 3 microM nicotine on the frequency of the spontaneous postsynaptic currents. This effect of 3 microM nicotine was also blocked by 20 microM bicuculline, a competitive antagonist of the GABA(A) receptor; in contrast, it was not affected by 1 mM kynurenic acid, an antagonist of the ionotropic glutamate receptor. In the presence of 1 microM tetrodotoxin, 3 microM nicotine was unable to affect the synaptic activity. Our findings suggest the existence of nicotinic receptors on GABAergic axons projecting to the vagal motoneurons. Because the effect is completely abolished by 1 microM tetrodotoxin, the nicotinic receptors are not localized on the presynaptic nerve terminal and their action on the GABA release requires the propagation of an action potential from their location to the synaptic terminal. This effect of nicotinic receptor activation on spontaneous GABA release in the dorsal motor nucleus of the vagus may have an important role in the regulation of gastrointestinal motility. PMID- 9219933 TI - Determination of NADH in the rat brain during sleep-wake states with an optic fibre sensor and time-resolved fluorescence procedures. AB - The present paper reports a nanosecond time-resolved fluorescence derived from the cortex and the area of the periaqueductal gray including the nucleus raphe dorsalis (PAG-nRD) in unanaesthetized freely moving rats. The measurements were acquired through a single optic fibre transmitting a subnanosecond nitrogen laser pulse (337 nm, 15 Hz) and collecting the brain fluorescence occurring at 460 nm which might depend on mitochondrial NADH (reduced form of nicotinamide adenine dinucleotide). The fluorometric method was combined with polygraphic recordings, and this procedure allowed us to define, for the first time, variations of the 460 nm signal occurring throughout the sleep-wake cycle. In the PAG-nRD, the signal exhibited moderate heterogeneous variation in amplitude during slow-wave as compared to the waking state. Constant increases were observed during paradoxical sleep as compared to the waking state. For this state of sleep the magnitude of the variations depended on the optic fibre location. In the cortex and during either slow-wave sleep or paradoxical sleep, the signal presented moderate increases which were significant during paradoxical sleep. The magnitude of the redox variations observed either in the PAG-nRD or in the cortex might be ascribed to the oxidative energy balance which is related to sleep states. PMID- 9219934 TI - The effects of donor stage on the survival and function of embryonic striatal grafts in the adult rat brain. I. Morphological characteristics. AB - The effects of the stage of donor embryos on the survival of grafts from different neuronal cell types have been well documented. Indeed, this parameter has been shown to be highly important in the survival and function of transplants of various tissues of the CNS. However this question has not been addressed in grafts of embryonic striatal tissue transplanted into animal models of Huntington's disease. In this study, rats which had received a unilateral ibotenic acid lesion in the dorsal striatum received grafts from a standard dissection of embryonic striatal primordium taken from donors of embryonic stage either E14, E16, E17 or E19 days. Three months after transplantation six rats from each group were killed for analysis of graft survival and morphology. The remaining animals in each group were killed between 10 and 14 months after grafting. Graft morphology was detected using a range of markers including: acetylcholinesterase and Cresyl Violet, the 32,000 mol. wt dopamine- and cyclic AMP-regulated phosphoprotein (DARPP-32), tyrosine hydroxylase and striatally enriched phosphatase. All the grafts from different donor stages survived well at both time-points and Cresyl Violet staining indicated neuronal cell types spread throughout the grafts. The transplants were seen to have a characteristic "patchy" appearance with areas of dense AChE activity and DARPP-32 immunopositivity interspersed with areas of much lighter expression. These areas also co-localized consistently with striatally-enriched phosphatase and tyrosine hydroxylase expression, indicating that they comprised the striatal-like compartment of the graft (the so called P zones, containing cells of the mature striatum), and receiving specific afferent input from the host dopaminergic system. There was no significant difference in total graft volume, when comparing individual groups at both time-points from grafting. However, when comparing the volume of the P zones, the striatal primordium from the youngest donor stages (E14 and E16) produced grafts with a significantly higher proportion of striatal like tissue. Therefore, in order to increase the proportion of striatal tissue within these grafts, tissue from younger embryonic donors should be used. This has important implications in the application of this model towards clinical trials in Huntington's disease. PMID- 9219935 TI - The effects of donor stage on the survival and function of embryonic striatal grafts in the adult rat brain. II. Correlation between positron emission tomography and reaching behaviour. AB - Grafts of embryonic striatal primordia are able to elicit behavioural recovery in rats which have received an excitotoxic lesion to the striatum, and it is believed that the P zones or striatal-like tissue within the transplants play a crucial role in these functional effects. We performed this study to compare the effects of different donor stage of embryonic tissue on both the morphology (see accompanying paper) and function of striatal transplants. Both the medial and lateral ganglionic eminence was dissected from rat embryos of either 10 mm, 15 mm, 19 mm, or 23 mm crown-rump length, and implanted as a cell suspension into adult rats which had received an ibotenic acid lesion 10 days prior to transplantation. After four months the animals were tested on the "staircase task" of skilled forelimb use. At 10-14 months rats from the groups which had received grafts from 10 mm or 15 mm donor embryos were taken for positron emission tomography scanning in a small diameter positron emission tomography scanner, using ligands to the dopamine D1 and D2 receptors, [11C]SCH 23390 and [11C]raclopride, respectively. A lesion-alone group was also scanned with the same ligands for comparison. Animals which had received transplants from the 10 mm donors showed a significant recovery with their contralateral paw on the "staircase test". No other groups showed recovery on this task. Similarly, the animals with grafts from the youngest donors showed a significant increase in D1 and D2 receptor binding when compared to the lesion-alone group. No increase in signal was observed with either ligand in the group which had received grafts from 15 mm donors. Success in paw reaching showed a strong correlation to both the positron emission tomography signal obtained and the P zone volume of the grafts. These results suggest that striatal grafts from younger donors (10 mm CRL) give greater behavioural recovery than grafts prepared from older embryos. This recovery is due to both the increased proportion of striatal-like tissue within the grafts and an increase in functional D1 and D2 dopamine receptors measured by positron emission tomography, i.e. a more extensive integration of the graft with the host brain. PMID- 9219936 TI - Glial and endothelial cell response to a fetal transplant of purified neurons. AB - Astrocytes, microglia and endothelial cells display very specific phenotypic characteristics in the intact adult CNS, which appear quite versatile when grown in culture without neurons. Indirect evidence from in vitro co-culture studies and analysis of the effects of specific neuronal removal in vivo, does accordingly favour a role of neurons for the phenotypic repression of these cells in the intact brain. In order to provide more direct evidence for such neuronal influence, we attempted to induce, in the rat brain, a reversal of the post lesional activation of astrocytes, microglia and endothelial cells by transplantation of fetal neurons purified by immunopanning. Host microglial cells which have been activated by the lesion process, penetrated the neuronal graft during the few days after the transplantation. Reactive astrocytes began to appear in the lesioned parenchyma and gathered around the transplant. Thereafter they first sent their processes in the direction of the neuronal graft, before they migrated into the graft a few days later. At this time, which was at the end of the first week post-transplantation, the host endothelial cells sprouted "streamers" of basal lamina within the graft forming small capillaries. During the second week post-transplantation, numerous astrocytes and microglial cells, both displaying a reactive hypertrophied morphology, were observed throughout the grafts. Finally, by the end of the first month, the activated cells differentiated towards a quiescent, resting morphology. At this time the grafts contained a vascular network with morphological characteristics comparable to those observed in the intact brain parenchyma. The results indicate that the interaction of activated astroglia and microglia and endothelial cells with neurons causes the cells to re-differentiate and regain phenotypic features characteristic of intact brain parenchyma, strongly suggesting that neurons play an essential role in the phenotypic restriction of glial and endothelial cells in the adult central nervous system. PMID- 9219937 TI - An in-frame deletion in peripheral myelin protein-22 gene causes hypomyelination and cell death of the Schwann cells in the new Trembler mutant mice. AB - Cloning and sequencing of the peripheral myelin protein-22 cDNA and genomic DNA from newly found Trembler mice revealed an in-frame deletion including exon IV which codes for the second (TM2) and a part of third (TM3) transmembrane domain of peripheral myelin protein-22. This mutation was distinct from those in both other allelic Trembler and Trembler-J mice, which carry point mutations within the putative transmembrane spanning regions of peripheral myelin protein-22. Inheritance was autosomal dominant. The affected mice revealed an abnormal gait, which appeared at 15-20 days of age, followed by motor and sensory ataxia, which remained throughout life. Most of the affected mice could survive more than one year. One of the most notable pathological phenotypes was a giant vacuolar formation in the sciatic nerve of homozygotes. They vary in size within the cytoplasm of Schwann cells, which failed to assemble myelin at any ages studied. Heterozygotes showed normal myelination during the early postnatal stages, followed by a segmental demyelination at an advanced stage. Vacuolar formation was not so frequent as in the homozygotes. These results suggest that the missing of transmembrane spanning region (TM2 and TM3) of peripheral myelin protein-22 may disturb a dual biological function of peripheral myelin protein-22, leading to a dysmyelination of axons and to a vacuolar formation within the cytoplasm of the Schwann cells. The latter phenotype is discussed in conjunction with the disruption of an intracellular transport system and subsequent cell death. PMID- 9219938 TI - Long-term glucocorticoid treatments decrease local cerebral blood flow in the rat hippocampus, in association with histological damage. AB - The present study examined the influence of a long-term treatment with glucocorticoid on local cerebral blood flow of the hippocampus in rats, estimated with the hydrogen clearance method. Either a cholesterol (100 mg, as a control) or corticosterone (100 mg) bead was implanted subcutaneously in rats for a period of three months, beginning at 12 weeks of age. The effects of the treatments on the local circulation of the hippocampus were evaluated three to four months after the termination of the treatments. Hippocampal cerebral blood flow in corticosterone-treated rats was significantly lower (P<0.05) than that in control rats, and fluctuated over a day in lower amplitude than the controls. Severe histological damage was observed in the CA1 and CA3 cell fields of the hippocampus in corticosterone-treated rats. These neuropathological changes were characterized by soma shrinkage and condensation, or nuclear pyknosis, as reported previously. We concluded that a long-term glucocorticoid exposure resulted in an impairment of the hippocampal functions, accompanied by neuronal damage similar to that found in aged hippocampus. The present results support the hypothesis that glucocorticoids accelerate age-related changes in the brain. PMID- 9219939 TI - Antagonism of adenosine A2A receptors underlies the behavioural activating effect of caffeine and is associated with reduced expression of messenger RNA for NGFI-A and NGFI-B in caudate-putamen and nucleus accumbens. AB - Caffeine, the most widely consumed of all psychostimulant drugs, exerts its action by antagonizing adenosine receptors. To study the arousing properties of caffeine, we injected rats intraperitoneally with vehicle, caffeine (7.5, 15 or 30mg/kg), the selective adenosine A2A receptor antagonist, SCH 58261 (3.75 mg/kg) or the selective adenosine A1 receptor selective antagonist DPCPX (7.5 mg/kg). In a behavioural test it was found that administration of caffeine and SCH 58261 significantly increased locomotion and rearing, whereas DPCPX did not alter locomotion and reduced rearing. After the behavioural session the rats were killed, their brains were cut at several levels along a rostrocaudal axis and in situ hybridization against NGFI-A messenger RNA and NGFI-B messenger RNA was performed. A reduction of NGFI-A messenger RNA was found in several subregions of both caudate putamen and nucleus accumbens in caffeine-treated animals. Similarly, animals that had received SCH 58261 showed significant decreases of NGFI-A messenger RNA in the rostral part of caudate putamen and in the shell part of nucleus accumbens. By contrast, DPCPX treatment caused an increase in the expression of NGFI-A messenger RNA and a smaller increase in NGFI-B messenger RNA in the lateral parts of caudate putamen. In addition, it was found that caffeine, but not SCH 58261 or DPCPX, elevated the expression of NGFI-A and NGFI-B messenger RNA in the cerebral cortex, especially in its parietal part. Thus, these results provide evidence that endogenous adenosine, via adenosine A2A receptors, causes a tonic activation of striatopallidal neurons. By blocking this adenosine effect, caffeine causes behavioural activation. PMID- 9219940 TI - Brain-derived neurotrophic factor promotes axonal regeneration and long-term survival of adult rat spinal motoneurons in vivo. AB - This study shows that in adult rat spinal motoneurons brain-derived neurotrophic factor exerts a neuroprotective effect which extends several weeks beyond the duration of treatment. In addition, brain-derived neurotrophic factor strongly enhances regeneration of avulsed motor axons across the border between the central and peripheral nervous systems. Treatment with brain-derived neurotrophic factor is known to rescue adult rat spinal motoneurons from retrograde cell death induced by ventral root avulsion. The present experiments were designed to test whether this survival effect remains over an extended period of time following cessation of treatment and, also, whether brain-derived neurotrophic factor promotes regeneration of avulsed motor axons. After avulsion of a spinal ventral root, four weeks of treatment with brain-derived neurotrophic factor (10 microg/day) or vehicle was initiated. By using different retrograde tracers to obtain pre- and postoperative labelling of avulsed and regenerating motoneurons, respectively, the number of surviving motoneurons as well as the extent of motor axonal regeneration could be analysed. The expression of nitric oxide synthase in the lesioned motoneurons was also studied. In the vehicle-treated rats, only 10% of the avulsed motoneurons remained at 12 weeks postoperatively, 20-40% of which displayed nitric oxide synthase activity. Treatment with brain-derived neurotrophic factor during the initial four postoperative weeks resulted in 45% motoneuron survival and a complete blockage of nitric oxide synthase expression at 12 weeks postoperatively. Brain-derived neurotrophic factor also induced abundant regeneration of the avulsed motor axons, which formed extensive fibre bundles along the surface of the spinal cord and adjacent ventral roots. The long term effect by brain-derived neurotrophic factor seemed to be even stronger on motor axonal regeneration than on motoneuron survival. The present results indicate a therapeutic potential for brain-derived neurotrophic factor in the early treatment of traumatic injuries to spinal nerves and roots. PMID- 9219941 TI - Basic fibroblast growth factor, its high- and low-affinity receptors, and their relationship to form-deprivation myopia in the chick. AB - Form deprivation myopia in chickens is a widely accepted model to study visually regulated postnatal ocular growth. Recently we showed that basic fibroblast growth factor-2 provides a "stop" signal for the growing eye. To understand further its action, we have localized basic fibroblast growth factor-2 and its low- and high-affinity receptors in the chicken eye, and determined the localization of basic fibroblast growth factor receptors in the inner plexiform layer with respect to that of neurotransmitter systems known to play a role in form-deprivation myopia. By immunocytochemistry and in situ hybridization, two complementary methods, we found that nearly all cells in the retina, and scleral chondrocytes, contain basic fibroblast growth factor-2 protein and messenger RNA as well as high-affinity basic fibroblast growth factor receptor protein and messenger RNA. Immunocytochemical localization of basic fibroblast growth factor 2 binding sites (a high resolution alternative to autoradiography), combined with N-glycanase and heparitinase treatment or heparin competition, revealed additional binding sites in specific synaptic layers of the inner plexiform layer and low-affinity binding sites in the choroid and optic fibre layer. Some binding sites in the synaptic layers were found to co-stratify with neurites of dopamine , vasoactive intestinal polypeptide- or enkephalin-containing amacrine cells, suggesting that basic fibroblast growth factor-2 could modulate synaptic transmission to or from these cells. Form deprivation did not affect the levels of basic fibroblast growth factor receptor-1 messenger RNA in retina/retinal pigment epithelium/choroid (Northern blotting), but it abolished the decrease in amount of extractable basic fibroblast growth factor normally observed in the dark (Western blotting). The results are discussed with respect to previous findings on basic fibroblast growth factor-2 and basic fibroblast growth factor receptor-1 localization in the avian and other vertebrate eyes, and their relevance to form-deprivation myopia. The widespread distribution of basic fibroblast growth factor-2 and its receptor makes it impossible to predict which cells might mediate the action of basic fibroblast growth factor-2 in form deprivation myopia. However, the alteration in amounts of extractable retinal basic fibroblast growth factor-2 in form-deprived, dark-adapted retinas, in which basic fibroblast growth factor-2 probably serves as a "stop" signal for ocular growth, is consistent with a role for basic fibroblast growth factor-2 in the regulation of ocular growth. PMID- 9219942 TI - Decreased skin sensory innervation in transgenic mice overexpressing insulin-like growth factor-II. AB - Cutaneous sensory innervation was studied in transgenic mice overexpressing insulin-like growth factor II using a keratin promoter. The skin area of these animals is enlarged providing increased target for sensory neurons. L4 dorsal root ganglion cell counts revealed that the total number of sensory neurons was the same in transgenics as control animals. Levels of nerve growth factor per unit weight of skin were also unchanged. The cutaneous nerves of the hindlimb were immunostained with the pan-neuronal marker PGP 9.5 in transgenic and control mice at postnatal day 0 and 21. The innervation in transgenic mice was markedly reduced, particularly in superficial dermis and epidermis and in some areas innervation was completely absent. The effect was greatest in distal skin regions and increased with age. Since insulin-like growth factor II has been reported to be a sensory neurotrophic factor, its effect on neurite outgrowth was tested on embryonic day 14 and 18 mouse lumbar dorsal root ganglion explants in culture. Under these conditions insulin-like growth factor II (5-100 ng/ml) did not have strong growth promoting activity and at embryonic day 18, in the presence of 5-10 ng/ml nerve growth factor, neurite outgrowth was suppressed by insulin-like growth factor II. The results show that increased skin target and availability of nerve growth factor per se do not alter the number of innervating sensory neurons. However, reduced sensory terminal arborization and skin hypoinnervation does occur in the presence of excess insulin-like growth factor-II. It is possible that insulin-like growth factor-II inhibits terminal axon growth directly via receptors on sensory neurons or peripheral glia. PMID- 9219943 TI - Neurotoxic consequences of central long-term administration of interleukin-2 in rats. AB - Interleukin-2 is an immunoregulatory cytokine with several recently established CNS activities. Central effects of interleukin-2 include growth promotion for neuronal and glial cells as well as modulatory influences on neurotransmission and hormone release. However, little is known about the consequences in the CNS of chronically elevated levels of interleukin-2. Alterations in the interleukin 2/interleukin-2 receptor system are not only associated with CNS trauma, inflammation and certain neuropathologies; elevated interleukin-2 concentrations are especially induced during the therapeutic use of interleukin-2 in cancer treatments. In the present study, intracerebroventricular (i.c.v.) interleukin-2 infusions (5 15 U/h) were performed in Sprague Dawley rats for up to 14 days. Interleukin-2-treated animals showed significantly increased plasma levels of corticosterone indicating an hyperfunctioning of the hypothalamic-pituitary adrenocortical axis that lasted over the 14 day infusion period. Moreover, the performance of interleukin-2-treated animals in the Morris swim maze task was transiently impaired. Quantitative receptor autoradiographic analyses revealed changes in the binding levels of cholinergic M1 and M2 as well as dopaminergic D1 and D2 receptors in selected brain areas in which interleukin-2 was shown to modulate neurotransmission and which are enriched with interleukin-2 receptor expression. Decreased receptor binding levels were observed in the frontoparietal cortex (M2, D1, D2), hippocampal CA1 region (M1, M2) and the nucleus accumbens (D2). Histological and immunohistochemical examination of the brains of interleukin-2-treated animals revealed multiple alterations. Interleukin-2 treatment resulted in an intracranial accumulation of non-neural, MHC class II positive cells as well as T and B lymphocytes within the infused brain hemisphere. Cellular infiltrates were associated with angiogenesis and the deposition of extracellular matrix material, such as fibronectin. Adjacent brain regions that were partly invaded and dislodged by the cellular masses were characterized by reactive astrogliosis, microglial activation, endothelial upregulation of adhesion molecules, myelin damage and neuronal loss. Together the data suggest that persistently elevated central levels of interleukin-2 can interfere with several CNS functions and may lead to nervous tissue injury. These findings could be relevant to CNS pathologies characterized by abnormal interleukin-2 production and to central responses to interleukin-2 treatments. PMID- 9219944 TI - Metabolic alterations produced by 3-nitropropionic acid in rat striata and cultured astrocytes: quantitative in vitro 1H nuclear magnetic resonance spectroscopy and biochemical characterization. AB - Quantitative high resolution in vitro 1H nuclear magnetic resonance spectroscopy was employed to study the metabolic effects of 3-nitropropionic acid associated with aging from perchloric acid extracts of rat striata. Systemic injection of 3 nitropropionic acid in rats at a dose of 10 mg/kg/day for seven consecutive days significantly impaired energy metabolism in rats one, four and eight months of age, as evidenced by a marked elevation of succinate and lactate levels. However, a significant decrease in N-acetyl-L-aspartate level, a neuronal marker, was observed in four- and eight-month-old rats but not in one-month-old rats. This would indicate that rats at four to eight months are more susceptible to 3 nitropropionic acid than those at one month. A significant decrease in GABA level was observed in four-month-old 3-nitropropionic acid-treated rats, which is consistent with the literature that GABAergic neurons are particularly vulnerable to 3-nitropropionic acid treatment. In addition, glutamine and glutamate levels were markedly decreased at four and eight months in 3-nitropropionic acid-treated rats. Since glutamine is synthesized predominantly in glia, the observation above suggests that 3-nitropropionic acid intoxication may involve perturbation of energy metabolism, glial injury and consequent neuronal damage. Astrocytes which are essential in the metabolism of glutamate and glutamine were used to further assess 3-nitropropionic acid-induced toxicity. Glial proliferation, mitochondrial metabolism and glutamine synthetase activity were all reduced by 3-nitropropionic acid treatment with a concomitant increase, in a dose-dependent manner, of lactate levels, suggesting that 3-nitropropionic acid is also detrimental to astrocytes in vivo and thus may affect metabolic interaction between neurons and glia. These results not only imply that 3-nitropropionic acid blocks energy metabolism prior to exerting neurotoxic damage but also demonstrate that the degree of energy depletion determines the detrimental effects of 3-nitropropionic acid. In the present study, we also demonstrate that glutamate and glutamine levels as well as astrocytic functions may play pivotal roles in 3-nitropropionic acid-induced striatal lesions. PMID- 9219945 TI - Cholinergic basal forebrain projections to nitric oxide synthase-containing neurons in the rat cerebral cortex. AB - Stimulation of basal forebrain neurons elicits regional cerebral blood flow increases which are reportedly mediated by acetylcholine and nitric oxide. However, the modality of interaction between these two mediators remains unclear. Particularly, little is known about the source, i.e. endothelial, glial and/or neuronal, of the potent gaseous vasodilator nitric oxide. In the present study, we examined, by double immunocytochemical labelling of nitric oxide synthase and choline acteyltransferase at the light and electron microscopic level, the existence of morphological relationships between cortical nitric oxide synthase containing neurons and cholinergic cells or nerve fibres. Using anterograde tract tracing and selective basal forebrain lesions, we further investigated the origin of the cholinergic input to cortical nitric oxide synthase neurons. The results confirm that cortical nitric oxide synthase-immunoreactive neurons are often associated with the local microvascular bed, show that intracortical neurons immunostained for nitric oxide synthase and choline acetyltransferase belong to two distinct neuronal populations and, further, that a subset of nitric oxide synthase-containing cell bodies and their proximal dendrites receive a cholinergic input which originates primarily from basalocortical projections. Altogether, these findings suggest that cholinergic basal forebrain neurons could increase cortical blood flow partly via a local nitric oxide relay neuron whereby the freely diffusing gas would be the direct smooth muscle vasodilator agent. It is concluded that this interaction might contribute to the complex relationships between the basal forebrain and the cortical microcirculation, interactions which result in fine regulation of cortical perfusion. PMID- 9219946 TI - Selective muscarinic antagonists differentially affect in vivo acetylcholine release and memory performances of young and aged rats. AB - Brain acetylcholine release and memory performance were investigated in young (three- to six-months) and old (20- to 24-months) rats. Acetylcholine release was measured in vivo in the cortex and hippocampus of freely-moving animals, under basal conditions and in the presence of the following muscarinic antagonists: scopolamine, (+/-)-5,11-dihydro-11-[[(2-[2-[(dipropylamino) methyl]-1 piperidinyl]ethyl) amino] carbonyl]-6H-pyrido(2,3-b)(1,4)-benzodiazepine-6-one (AFDX 384) and pirenzepine. The amount of acetylcholine released from the cortex and hippocampus of old rats was significantly reduced. In the presence of scopolamine and AFDX 384 but not of pirenzepine, the acetylcholine release was significantly higher in the old than the young rats, suggesting that changes in presynaptic M2/M4 muscarinic receptor function occur with ageing in the two brain regions. Cognitive capacities were evaluated using two different behavioural tasks: object recognition and passive avoidance response. Old rats were unable to discriminate between familiar and novel objects and had impaired performance in the passive avoidance test. AFDX 384 restored the performance in both tests. Furthermore, in young rats AFDX 384 reversed the impairment of both object recognition and passive avoidance response induced by scopolamine. The effect of AFDX 384 on acetylcholine release and behaviour in the old rats offers further support to a relationship between the age-related cholinergic hypofunction and cognitive impairment and indicates the blockade of presynaptic muscarinic receptors as a possible selective target for therapeutic strategies aimed at improving age-associated memory deficits. PMID- 9219947 TI - Enhanced taurine release in cell-damaging conditions in the developing and ageing mouse hippocampus. AB - Taurine has been shown to be essential for neuronal development and survival in the central nervous system. The release of preloaded [3H]taurine was studied in hippocampal slices from seven-day-, three-month- and 18-22-month-old mice in cell damaging conditions. The slices were superfused in hypoxic, hypoglycemic and ischemic conditions and exposed to free radicals and oxidative stress. The release of taurine was greatly enhanced in the above conditions in all age groups, except in oxidative stress. The release was large in ischemia, particularly in the hippocampus of aged mice. Potassium stimulation was still able to release taurine in cell-damaging conditions in immature mice, whereas in adult and aged animals the release was so substantial that this additional stimulus failed to work. Taurine release was partially Ca2+-dependent in all cases. The massive release of the inhibitory amino acid taurine in ischemic conditions could act neuroprotectively, counteracting in several ways the effects of simultaneous release of excitatory amino acids. This protection could be of great importance in developing brain tissue, while also having an effect in aged brains. PMID- 9219948 TI - Inducible expression of N-methyl-D-aspartate receptor, and delta and mu opioid receptor messenger RNAs and protein in the NT2-N human cell line. AB - Retinoic acid treatment of NT-era2/cl.D1 (NT2) cells, a human teratocarcinoma cell line, yields 95% pure cultures of terminally differentiated neuronal cells. Concomitant with their terminal differentiation into neurons, NT2 cells are induced by retinoic acid to express neuronal N-methyl-D-aspartate receptor channels, which are fully functional. We determined the effects of retinoic acid induced differentiation of NT2 cells on the levels of N-methyl-D-aspartate, delta opioid and mu opioid receptor messenger RNAs. RNA levels were measured using quantitative solution hybridization assays. The riboprobes were complementary to major portions of the coding regions of the N-methyl-D-aspartate, delta opioid and mu opioid receptor complementary DNAs. After four weeks of exposure to 10 microM retinoic acid, followed by four weeks of treatment with mitotic inhibitors (1 microM of cytosine arabinoside, 10 microM of fluorodeoxyuridine and 10 microM of uridine) the levels of N-methyl-D-aspartate receptor messenger RNA in differentiated NT2-N cells increased 10-fold, delta opioid receptor messenger RNA increased three-fold, and mu opioid receptor messenger RNA increased four-fold. Northern blot analysis revealed two transcripts for the N-methyl-D-aspartate receptor messenger RNA (4.2 and 4.4 kb) and two transcripts for delta opioid receptor messenger RNA (7.0 and 11.0 kb). To determine whether the increases in messenger RNAs were accompanied by an increased synthesis of the respective proteins, we examined the immunoperoxidase localization of N-methyl-D-aspartate receptor and delta opioid receptor antisera. N-Methyl-D-aspartate receptor-like immunoreactivity was seen within the cell bodies as well as on the processes of the retinoic acid-differentiated cells. Although delta opioid receptor-like immunoreactivity was detected within the soma of isolated cells prior to retinoic acid treatment, the apparent number of these labelled cells and their ramified processes were markedly enhanced following retinoic acid differentiation. These results demonstrate parallels between the inducible expression of the N-methyl-D aspartate and opioid receptor messenger RNAs and proteins during the acquisition of the fully differentiated neuronal phenotype in cultured NT2 cells. Retinoic acid-differentiated NT2 cells express increased levels for the N-methyl-D aspartate, delta opioid and mu opioid receptor messenger RNAs, providing the opportunity to study the interactions among these receptor systems in human terminally differentiated neuronal cells in culture. PMID- 9219949 TI - A ventrodorsal GABA gradient in the embryonic retina prior to expression of glutamate decarboxylase. AB - GABA is known to function as a neurotransmitter in the mature nervous system, and in immature neurons it has been linked to neurotrophic actions. While most GABA is generated by glutamate decarboxylase (GAD), an alternative synthetic pathway is known to originate from putrescine, which is converted via gamma aminobutyraldehyde in an aldehyde-dehydrogenase-requiring step to GABA. In a search for the role of two aldehyde dehydrogenases expressed in segregated compartments along the dorsoventral axis of the developing retina, we assayed dorsal and ventral retina fractions of the mouse for GABA by high performance liquid chromatography. We found GABA to be present in the embryonic retina, long before expression of GAD, and ventral GABA levels exceeded dorsal levels by more than three-fold. Postnatally, when GAD became detectable, overall GABA levels increased, and the ventrodorsal concentration difference disappeared. Our observations indicate that prior to the formation of synapses the embryonic retina contains a ventrodorsal GABA gradient generated by an alternate synthetic pathway. PMID- 9219950 TI - Subdivisions of the guinea-pig accessory olfactory bulb revealed by the combined method with immunohistochemistry, electrophysiological, and optical recordings. AB - The presence of subgroups in vomeronasal sensory neurons has been known in various animals. To elucidate possible functional subdivisions in the guinea-pig accessory olfactory bulb, the combined studies with GTP-binding protein immunohistochemistry, electrophysiological and optical recordings were carried out. Gi2 alpha and Go alpha proteins were immunohistochemically localized, respectively, in the anterior and posterior regions of the vomeronasal nerve and glomerular layers, indicating that the guinea-pig accessory olfactory bulb receives at least two different inputs. This suggests that an anatomical boundary exists in these two layers. A mapping study of field potentials in sagittal slice preparations demonstrated that stimulation of the anterior vomeronasal nerve layer elicited field potentials with weak oscillatory responses exclusively in the anterior region of the external plexiform layer, whereas shocks to the posterior vomeronasal nerve layer provoked distinct oscillatory responses within the posterior one. The damping factors of oscillations in the anterior and posterior regions were 0.064+/-0.028 and 0.025+/-0.014, respectively. These electrophysiological results suggest that the accessory olfactory bulb consists of two functionally different subdivisions. Real-time optical imaging showed that anterior vomeronasal nerve layer shocks produced neural activity which spread horizontally from anterior to posterior only within the anterior region of the external plexiform and mitral cell layers, whereas shocks to the posterior vomeronasal nerve layer evoked periodic neural activity which spread horizontally from posterior to anterior only within the posterior region. Furthermore, the most posterior extent of the optical response evoked in the anterior region immediately adjoined the most anterior extent of that evoked in the posterior region. The maximal distance of signal propagation in the granule cell layer corresponded to that in the overlying external plexiform and mitral cell layers, indicating that the granule cell layer also has a similar boundary. Thus, these optical imaging studies not only demonstrated a precise boundary in each layer of the accessory olfactory bulb, which was positioned right beneath the boundary defined by GTP-binding protein immunohistochemistry, but also confirmed the observations from electrophysiological mapping that evoked field potentials are independently distributed in each of two subdivisions. The presence of the functional subdivision in each layer leads us to conclude that the accessory olfactory bulb in the guinea-pig is distinctly segregated into the anterior and posterior subdivisions, and to suggest that there are at least two different input output pathways in the vomeronasal system. PMID- 9219951 TI - Non-conventional role of lysosomal acid phosphatase in olfactory receptor axons: co-localization with growth-associated phosphoprotein-43. AB - Olfactory receptor neurons undergo a continuous turnover in adult mammals. It is largely unknown how their axons invade the olfactory bulb and induce synaptic re organization in glomeruli. Here, the cytochemical localization of lysosomal acid phosphatase has been studied in olfactory bulbs of adult rats and mice. The enzyme has been identified by specific substrate, inhibitors and absence in lysosomal acid phosphatase-knockout mice. Lysosomal acid phosphatase is located in primary and secondary lysosomes, which are unevenly distributed in the olfactory nerve layer and among olfactory glomeruli. In consecutive sections of glomeruli, the intensity of lysosomal acid phosphatase immunoreactivity co-varied with that of growth-associated phosphoprotein. Electron microscopically, differential lysosomal acid phosphatase staining in glomeruli corresponded to different proportions of labelled and unlabelled axons. Quantification revealed that lysosomal acid phosphatase labelling was strongest in non-synaptic profiles of terminal axons, while it was weak in or even missing from most synaptic profiles. Hence, growing olfactory axons apparently carry more lysosomal acid phosphatase than those which have established synaptic contacts. Following olfactory deafferentation both lysosomal acid phosphatase activity and growth associated phosphoprotein-43 are lost from glomeruli, suggesting that both proteins are expressed in olfactory sensory axons during growth, while lysosomal acid phosphatase is apparently not a marker of anterograde terminal degeneration. PMID- 9219952 TI - Studies on the release and extracellular metabolism of endogenous ATP in rat superior cervical ganglion: support for neurotransmitter role of ATP. AB - The release of endogenous ATP, measured by the luciferin-luciferase assay, and the release of [3H]acetylcholine from the isolated superior cervical ganglion of the rat loaded with [3H]choline were studied simultaneously. Electrical field stimulation enhanced the release of endogenous ATP and acetylcholine in a [Ca2+]o dependent manner. The Na+ channel blocker, tetrodotoxin (1 microM) inhibited the stimulation-evoked release of endogenous ATP and of [3H]acetylcholine, but did not change the resting release. The release of ATP was dependent on the frequency of stimulation between 2 and 10 Hz. when the number of shocks was kept constant (360 shocks), while acetylcholine was not released in a frequency-dependent fashion. Ten days after cutting of the preganglionic nerve of the superior cervical ganglion the stimulation-evoked release of acetylcholine and ATP was abolished and the uptake of [3H]choline was significantly reduced but not inhibited. Hexamethonium, (100 microM) a nicotinic acetylcholine receptor antagonist, significantly reduced the release of both acetylcholine and ATP, indicating a positive feedback modulation of ACh and ATP release. 8-Cyclopentyl 1,3-dipropylxanthine (10 nM), the selective A1-adenosine receptor antagonist exhibited similar effect on the release of ATP and acetylcholine: both of them were augmented, showing that the stimulation-evoked release of ATP and acetylcholine are under the inhibitory control of A1-adenosine receptors. When the temperature was reduced to 7 degrees C to inhibit carrier-mediated processes, the resting and stimulated release of acetylcholine was not changed. Conversely, the release of ATP in response to stimulation was reduced by 79.9 +/- 5.6%, and the basal release was also almost completely blocked. Carbamylcholine by itself was able to release ATP, but not acetylcholine, in a hexamethonium-inhibitable manner, even from ganglia whose preganglionic nerve had been cut 10 days prior to experiments, suggesting that ATP release can occur in response to nicotinic receptor stimulation of postsynaptic cells. The breakdown of ATP or AMP by superior cervical ganglion was measured by high performance liquid chromatography combined with UV detection. ATP and AMP, added to the tissues, were readily decomposed: the Km (apparent Michaelis constant) and Vmax (apparent maximal velocity) were 475 +/- 24 microM and 3.50 +/- 0.18 nmol/min per mg for ectoATPase and 1550 +/- 120 microM and 14.5 +/- 0.9 nmol/min per mg tissue for 5' nucleotidase. In addition, by using electron microscopic enzyme histochemistry, the presence of ectoATPase was also shown in the superior cervical ganglion. It is concluded that endogenous ATP and acetylcholine are released simultaneously in response to stimulation of preganglionic nerve terminals in the superior cervical ganglion in a [Ca2+]o-dependent, tetrodotoxin-sensitive manner and is metabolized by ectoenzymes present in the tissue. The dissociation of the release of ATP and acetylcholine at different stimulation frequencies and temperatures shows that the release-ratio of acetylcholine and ATP can vary upon the condition of stimulation: this can reflect either the different composition of synaptic vesicles in the preganglionic nerve terminals or a significant contribution of non-exocytotic, carrier-mediated type of release of ATP to the bulk release. PMID- 9219953 TI - Characterization of cholecystokininA and cholecystokininB receptors expressed by vagal afferent neurons. AB - The cholecystokinin receptors expressed by vagal afferent neurons mediate the effect of cholecystokinin in inhibiting food intake and gastric emptying. We have determined the relative abundance of cholecystokininA, gastrin-cholecystokininB and gastrin-cholecystokininC receptor populations in the rat vagus by autoradiography using [125I]Bolton Hunter-cholecystokinin-8, [125I]Bolton Hunter heptadecapeptide gastrin and [125I]Leu(15)2-17Glycine-extended heptadecapeptide gastrin, together with the selective antagonists devazepide and L-740093. The results indicate approximately three-fold higher abundance of cholecystokininA compared with gastrin-cholecystokininB receptors, and no significant representation of gastrin-cholecystokininC receptors. Topical capsaicin applied to the vagal nerve trunk abolished the accumulation of sites binding both [125I]Bolton Hunter-labelled cholecystokinin-8 and heptadecapeptide gastrin indicating that both cholecystokininA and gastrin-cholecystokininB receptor populations were present on afferent fibres. The molecular identity of the receptors expressed by rat and human nodose ganglia was examined using the reverse transcription polymerase chain reaction. Products of the predicted size for the cholecystokininA and gastrin-cholecystokininB receptors were identified. The human and rat cholecystokininA receptor products were cloned and the sequences were found to be 99% homologous to those published for receptors expressed by rat pancreas and human gall bladder. We conclude that cholecystokininA and gastrin-colecystokininB receptors are synthesized by nodose ganglion cells, and that the receptor proteins are transported to the periphery along afferent fibres. While there is a clear role for vagal cholecystokininA receptors, the function of vagal afferent gastrin-cholecystokininB receptors remains to be determined. PMID- 9219954 TI - Expression of cholecystokinin messenger RNA in reciprocally-connected auditory thalamus and cortex in the rat. AB - Cholecystokinin exerts a potent antiepileptic action in mammalian auditory system and undergoes seizure-mediated up-regulation. The present study investigated cholecystokinin messenger RNA expression in the reciprocally-connected auditory thalamus and cortex in the rat. Immunofluorescence in situ hybridization was performed using a 24-base cholecystokinin-messenger RNA oligonucleotide probe. Corticothalamic projection neurons were identified by means of the retrograde fluorescent tracer rhodamine latex microspheres injected into the medial geniculate body. In our experiments, cholecystokinin messenger RNA transcripts were found in about 80% of neurons located within the reciprocally-connected regions of the medial geniculate body and the auditory cortices. These observations provide evidence of cholecystokinin production in the reciprocally connected regions of the auditory thalamus and cortex, the structures which jointly create the thalamo-corticothalamic circuit which has been implicated in seizure genesis. PMID- 9219955 TI - Movement-related and steady-state electromyographic activity of human elbow flexors in slow transition movements between two equilibrium states. AB - The electromyograms were recorded in healthy human subjects by surface electrodes from the mm. biceps brachii (caput longum et. brevis), brachioradialis, and triceps brachii (caput longum) during slow transition movements in elbow joint against a weak extending torque. The test movements (flexion transitions between two steady-states) were fulfilled under visual control through combining on a monitor screen a signal from a joint angle sensor with a corresponding command generated by a computer. Movement velocities ranged between 5 and 80 degrees/s, subjects were asked to move forearm without activation of elbow extensors. Surface electromyograms were full-wave rectified, filtered and averaged within sets of 10 identical tests. Amplitudes of dynamic and steady-state components of the electromyograms were determined in dependence on a final value of joint angle, slow and fast movements were compared. An exponential-like increase of dynamic component was observed in electromyograms recorded from m. biceps brachii, the component had been increased with movement velocity and with load increment. In many experiments a statistically significant decrease of static component could be noticed within middle range of joint angles (40-60 degrees) followed by a well expressed increment for larger movements. This pattern of the static component in electromyograms could vary in different experiments even in the same subjects. A steady discharge in m. brachioradialis at ramp phase has usually been recorded only under a notable load. Variable and quite often unpredictable character of the static components of the electromyograms recorded from elbow flexors in the transition movements makes it difficult to use the equilibrium point hypothesis to describe the central processes of movement. It has been assumed that during active muscle shortening the dynamic components in arriving efferent activity should play a predominant role. A simple scheme could be proposed for transition to a steady-state after shortening. Decrease of the efferent inflow can evoke internal lengthening of the contractile elements in muscle and, as a result, hysteresis increase in the muscle contraction efficiency. Effectiveness in maintenance of the steady position seems to also be enhanced due to muscle thixotropy and friction processes in the joint. Hysteresis after-effects in elbow flexors were demonstrated as a difference in steady-state levels of electromyograms with oppositely directed approaches to the same joint position. PMID- 9219956 TI - Effects of cutaneous afferent input on fatigue-induced changes in fusimotor activity of decerebrate cats. AB - Interaction of cutaneous and small-diameter, primarily fatigue-induced, muscle afferent inputs on fusimotor neurons has been studied in decerebrate cats. Spike discharges of fusimotor neurons to medial gastrocnemius were recorded from filaments dissected free from this muscle nerve. Non-noxious mechanical stimuli (10 Hz, 2 mm vibration) were applied to the skin area on the lateral side of the heel, innervated by sural nerve, during long-lasting (250 s) fatiguing contraction of lateral gastrocnemius and soleus muscles, elicited by electrical stimulation (40 Hz, 1.3 x motor threshold) of the muscle nerves. In 15 units (58%) the pattern of responses to muscle contraction and/or fatigue (initial transient, and late long-lasting increase in firing rate, respectively) was preserved in the presence of skin vibration which, by itself, provoked either a slight increase or no changes in fusimotor discharge rate. Pattern of the response to skin vibration prevailed in the presence of muscle contraction and fatigue only if the vibration by itself induced marked increase in fusimotor discharge rate (three units). In the remaining eight units the responses to both stimuli applied simultaneously were dissimilar in pattern to the response to either stimulus applied alone: the initial, tension-related, increase in firing rate was prolonged, while the late, fatigue-induced one was attenuated and its post-contraction part almost abolished. Possible mechanisms and functional role of interaction between cutaneous and muscle afferent inflows are discussed. PMID- 9219957 TI - Changes in dorsal neck muscle activity related to imposed eye movement in the decerebrate pigeon. AB - Movements of the head and eyes are known to be intimately related. Eye position has also been shown to be closely related to the electromyographic activity of dorsal neck muscles; however, extraocular muscle proprioception has not generally been considered to play a part in the control of such movements. We have previously shown that, in the pigeon, imposed movements of one eye modify the vestibular responses of several dorsal neck muscles in ways that are dependent on stimulus parameters such as the amplitude and velocity of imposed eye movement. The present study examines more closely the interactions between imposed eye movements and different muscle pairs. The three neck muscle pairs studied each responded to afferent signals from the extraocular muscles in discrete and specific ways which appeared to be correlated with their different actions. Complementary effects of imposed eye movements in the horizontal plane were seen for both the complexus and splenius muscle pairs, with imposed eye movements in one direction producing the largest inhibition of the ipsilateral muscle's vestibular response and imposed eye movements in the opposite direction the largest inhibition of the contralateral muscle's vestibular response. During roll tilt oscillation (ear-up/ear-down) in the frontal plane, similar complementary effects of imposed eye movement were seen in the complexus muscle pair, but the splenius muscle pair showed little tuning, with similar inhibition for imposed eye movement directed either upwards or downwards. In contrast to these complementary effects, the biventer cervicis muscle pair showed no vestibular modulation during vestibular stimulation in the horizontal plane and their spontaneous activity was not altered by imposed eye movement. During roll-tilt oscillation (ear-up/ear-down) in the frontal plane imposed eye movement directed vertically upwards increased both muscles' vestibular responses and imposed eye movement directed vertically downwards inhibited both muscles' vestibular responses. Section of the ophthalmic branch of the trigeminal nerve (deafferenting the eye muscles) abolished the effects of imposed eye movement on the neck muscle pairs. In conjunction with further control experiments these results provide compelling evidence that proprioceptive signals from the extraocular muscles reach the neck muscles and provide them with a functionally significant signal. We have previously shown that signals from the extraocular muscles appear to be involved in the control of the vestibulo-ocular reflex. It follows from the experiments reported here that proprioceptive signals from the extraocular muscles are also likely to be involved in the control of gaze. PMID- 9219959 TI - The effect of nigral implantation on sensitization to dopamine agonists in 6 hydroxydopamine-lesioned rats. AB - The implantation of fetal nigral tissue into the striatum of patients with Parkinson's disease is a promising approach to treatment which may produce clinical benefit partly by influencing drug responsiveness. The purpose of the present study was to determine the pharmacological mechanisms which drug response changes by measuring to what extent sensitization produced by repeated apomorphine treatment was attenuated by tissue implantation in rats with nigrostriatal lesions. Prior to implantation of nigral cell suspensions, the daily administration of apomorphine to rats with unilateral 6-hydroxydopamine lesions produced a progressive increase in the magnitude and duration of rotational behaviour. After implantation, apomorphine-induced rotational effects were reduced to levels observed upon the initial exposure to drug and did not increase following repeated treatment. Attenuated responses to selective D1 and D2 agonists were also observed after implantation. In vehicle-implanted rats, the initial response to apomorphine was attenuated but then increased following repeated apomorphine administration. No attenuation in responses to selective D1 and D2 agonists was observed in this group. Cell suspensions prepared from fresh and cyropreserved tissue produced similar behavioural effects, even though the volume of transplanted striatum exhibiting tyrosine hydroxylase activity was greater with fresh tissue. The duration of rotational behaviour induced by apomorphine was not affected by cell implantation. These findings suggest that the expression of sensitization in an animal model of parkinsonism may disappear after a period without drug treatment. Implantation of nigral tissue may produce beneficial results in parkinsonism by limiting the development of dopamine agonist-induced sensitization. PMID- 9219958 TI - The degree of inhibition of dopaminergic neurons in the ventral tegmental area induced by selective serotonin reuptake inhibitors is a function of the density power-spectrum of the interspike interval. AB - Electrophysiological techniques and computational methods were used to study the effect of the selective serotonin reuptake inhibitors fluvoxamine, paroxetine and sertraline on the basal activity of dopamine neurons in the ventral tegmental area. Acute injection of fluvoxamine, paroxetine and sertraline (20-1280 microg/ kg, i.v.) caused a dose-dependent inhibition of some ventral tegmental area DA neurons but it did not affect the basal firing rate of other DA cells. A Fast Fourier-Transformation based analysis of the basal activity of 32 ventral tegmental area DA neurons showed a positive correlation between the value of a functional operator (psi) equivalent to the density-power-spectrum of the signals and the degree of selective serotonin reuptake inhibitor-induced inhibition of ventral tegmental area DA cells. All ventral tegmental area DA neurons sampled were subdivided into two subclasses: (A) neurons with no changes in their basal firing rate and (B) neurons showing an approximately linear inhibitory effect in response to selective serotonin reuptake inhibitors. The neurons belonging to subclass A showed a more regular behavior of the interspike interval functions corresponding to lower values detected by the functional operator psi, whereas the neurons belonging to subclass B showed a less regular behavior of interspike interval functions corresponding to higher psi values detected by the same functional operator. Fluvoxamine, paroxetine and sertraline also caused a dose dependent increase of the percentage of spikes occurring in bursts in neurons belonging to subclass A (low values of psi), whereas the mean basal firing rate of these cells was not affected. It is suggested that this difference in density power-spectrum could reflect the asymmetry of serotonergic input to the ventral tegmental area DA neurons, and the differential effects of selective serotonin reuptake inhibitors on these neurons might depend on the characteristics of their basal firing mode. PMID- 9219960 TI - Decreased expression of N-methyl-D-aspartate receptor 1 messenger RNA in select regions of Alzheimer brain. AB - An antisense oligonucleotide probe was used to examine the expression of gene encoding the obligatory NMDAR1 subunit of the N-methyl-D-aspartate receptor in the hippocampus and adjacent cortical areas (entorhinal and perirhinal cortices) of seven Alzheimer patients and in the same brain regions of seven control individuals. Both groups were matched according to age, sex, cause of death, post mortem delay, and tissue storage time. Densitometric analysis of in situ hybridization autoradiograms revealed a 34% (P<0.05) decrease in NMDAR1 messenger RNA levels in layer III of the entorhinal cortex in Alzheimer brains. Similar deficits. although statistically not significant, were observed in layers II and IV-VI of the entorhinal cortex, and in granule cells of the dentate gyrus. Reduced levels of NMDAR1 messenger RNA were also found in layers II-VI of the perirhinal cortex (41 53% decrease, P<0.02). There were no changes in NMDAR1 messenger RNA expression in the CA1, hilus, or subiculum. Both Alzheimer and control group show substantial intersubject variation in levels of NMDAR1 messenger RNA. The analysis of emulsion-dipped tissue revealed a trend toward a decrease in the number of silver grains overlying individual neurons in the CA1, entorhinal cortex, and granule cell layer of some Alzheimer patients. No significant relationship was detected between the levels of NMDAR1 messenger RNA and post mortem delay, tissue storage, age of the subjects, or mini mental state exam score either in control or Alzheimer individuals. In contrast, a strong inverse correlation between NMDAR1 expression and disease duration was found. These data suggest that reduction in expression of the NMDAR1 gene observed in certain regions of Alzheimer hippocampus and adjacent cortical regions is specific for the disease itself. We postulate that reduced transcript levels may reflect either regional cell loss or anomalies in glutamatergic input to the hippocampus and entorhinal cortex in Alzheimer's disease. When followed by changes at the receptor subunit protein level, altered expression of the NMDAR1 gene in Alzheimer brain may contribute, through the formation of N-methyl-D aspartate receptors with different properties, to the previously reported modified N-methyl-D-aspartate receptor ligand binding, abnormal vulnerability of select neuronal populations to excitotoxic insult, and may also be involved in learning and memory deficits. PMID- 9219961 TI - The cerebral cortex is damaged in chronic alcoholics. AB - There is some controversy in the literature concerning whether chronic alcohol consumption damages the cerebral cortex. While decreased neuronal density in specific cortical regions is well described in chronic alcoholics, a recent study by Badsberg Jensen and Pakkenberg using unbiased stereological methods questions whether neurodegeneration occurs. In order to assess selective neurodegeneration in the cerebral cortex of chronic alcoholics, regional volumes and unbiased estimates of regional neuronal number (including neuronal identification with calcium-binding proteins) were calculated for 14 chronic alcoholics and 21 controls. Cases were carefully screened to exclude any interfering pathologies. Lifetime and maximum daily alcohol consumption was determined, and homogeneous groups were identified (four chronic alcoholics with Wernicke's encephalopathy and Korsakoff's psychosis, four chronic alcoholics with Wernicke's encephalopathy alone, six chronic alcoholics without Wernicke's encephalopathy or Korsakoff's psychosis, and 21 controls). Brain volume analysis revealed that discrete regions were significantly smaller in the chronic alcoholics compared to controls. As previously shown, white matter regions (particularly in the frontal lobe) were the most significantly reduced in volume. Alcoholics with Wernicke's encephalopathy (either alone or in combination with Korsakoff's psychosis) had significantly smaller white matter volumes than controls or alcoholics without these complications. Medial temporal lobe regions and the thalamus were also reduced in volume. Regression analyses revealed that the volume of both the white matter and thalamus negatively correlated with alcohol consumption. Consistent with the interpretation of previous neuronal density studies, selective neuronal loss was found in the superior frontal association cortex of chronic alcoholics, while no loss occurred from the motor cortex. The number of parvalbumin-, calbindin- and calretinin-immunoreactive neurons was found to be unaltered in chronic alcoholics, suggesting that the neurodegeneration is confined to the non GABAergic pyramidal neurons. As neurodegeneration was observed in all alcoholic groups, damage to the frontal association cortex is not restricted to alcoholics with the amnesia of Korsakoff's psychosis. These results are consistent with the notion that chronic alcohol consumption is associated with selective neuronal vulnerability. The selective frontal neurodegeneration and the frontal focus of white matter atrophy are supported by neuropsychological, regional blood flow, and magnetic resonance imaging studies of frontal lobe dysfunction in chronic alcoholics and may correlate with abnormalities in working memory. PMID- 9219962 TI - Apoptosis mediates cell death following traumatic injury in rat hippocampal neurons. AB - A model of in vitro traumatic injury with dissociated rat hippocampal neurons was studied to explore the mechanism of cell death. The neurotoxicity induced by traumatic injury to the cell culture can be transferred to a naive uninjured culture by media exchange. This toxicity is attenuated by dimethylsulfoxide or superoxide dismutase, suggesting that this toxicity is mediated by a free radical generation. Ionotropic glutamate receptor antagonists had no effect. This toxicity was effectively blocked by the pretreatment of the naive uninjured recipient cultures with cycloheximide or with actinomycin D. The DNA fragmentation could be illustrated with in situ nick translation in the cells which seem to have lost their cytoplasm. The nuclear morphology of neurons labeled by a neurofilament-specific antibody, SMI-31, demonstrated chromatin condensation and nucleosome formation. Traumatic injury induces release of an unknown toxin into the extracellular space. These observations suggest that a traumatized neuronal culture can propagate cell death of naive uninjured cells by releasing a neurotoxin that causes apoptosis. PMID- 9219963 TI - Effect of increased maternal corticosterone during lactation on hippocampal corticosteroid receptors, stress response and learning in offspring in the early stages of life. AB - The influence of maternal corticosterone during lactation on the development of the hippocampal corticosteroid receptor system, hypothalamus-pituitary-adrenal axis activity and spatial learning/retention performance was investigated in the rat during postnatal days 11 to 30. We increased the plasma levels of corticosterone by adding the hormone (200 microg/ml) to the drinking water of the dams. When compared to controls corticosterone-nursed offspring displayed: i) higher number of hippocampal type I and type II corticosteroid receptors at 30 days of life, but no changes at 11 and 16 days; ii) higher plasma levels of corticosterone in the basal condition and after 15 min of maternal separation at 11 but not at 16 days: iii) lower adrenal weights at 11 and 16 days, but which were no longer present at the age of 30 days; iv) no difference in performance in the place learning version of the Morris water task and T aquatic maze at 16 days. The present results, together with our previous findings showing that 90 day-old corticosterone-nursed rats have lower basal and restraint stress corticosterone levels and improved learning performance, indicate that the effects of maternal treatment appears only after weaning, thereby suggesting that increased corticosteroid receptors may be responsible, at least partially, for the endocrine and learning modifications induced by pre-weaning corticosterone exposure. The role played by maternal circulating corticosterone during the period of lactation in shaping the characteristics of the hypothalamus-pituitary adrenal axis and brain of the offspring is outlined. PMID- 9219964 TI - Intracellular calcium redistribution accompanies changes in total tissue Na+, K+ and water during the first two hours of in vitro incubation of hippocampal slices. AB - Changes of total tissue water, Ca, Na and K contents were monitored in whole transverse hippocampal slices of the guinea-pig during the first 2 h of in vitro incubation. A brief, 75% increase in tissue Ca was noted during the initial 15 min of maintenance, in contrast to a permanent increase of sodium and water contents, coupled to simultaneous decrease of potassium level. The rate of tissue Na, K and water changes comprised a rapid phase at the first 10-20 min, parallel with the increase of the tissue Ca content, and a slow phase during the rest of the incubation period. Development of specific morphological alterations, representative of ischemic/hypoxic lesions and a translocation of calcium from cytoplasm to mitochondria and endoplasmic reticulum during slice maintenance, was also detected by electron microscopy. A two-step mechanism might explain the development of a new steady-state total calcium content of slices. in which the cellular Ca2+ uptake at the beginning of incubation, likely triggered by hypoxic/ ischemic trauma of slice preparation, is followed by a balanced Ca2+ influx, extrusion and sequestration (predominantly into mitochondria and endoplasmic reticulum) during maintenance. PMID- 9219965 TI - Positive correlation between prolonged potentiation of binding of double-stranded oligonucleotide probe for the transcription factor AP1 and resistance to transient forebrain ischemia in gerbil hippocampus. AB - Gel retardation electrophoresis revealed that binding of a radiolabelled double stranded oligonucleotide probe for the nuclear transcription factor activator protein-1 was markedly potentiated in the CA1 and CA3 subfields and the dentate gyrus of the hippocampus of the gerbils with transient forebrain ischemia for 5 min, which is known to induce delayed death of pyramidal neurons exclusively in the CA1 subfield. The potentiation was transient in the vulnerable CA1 subfield, but persistent up to 18 h in the resistant CA3 subfield and dentate gyrus. However, no significant alteration was detected in endogenous levels of cyclic AMP response element binding protein phosphorylated at serine133 in these three different hippocampal structures 3 h after the reperfusion. On the other hand, hypothermia during ischemia which is known to protect the CA1 subfield against ischemic damages, led to a prolonged elevation of the activator protein-1 binding up to 9 h after the reperfusion in this vulnerable subfield at least in part through expression of c-Fos protein. Moreover, activator protein-1 binding was significantly elevated in the CA1 subfield up to 12 h after forebrain ischemia for 2 min which is shown not to induce marked damages to the vulnerable subfield. These results suggest that prolonged elevation of DNA binding activity of activator protein-1 may be responsible for molecular mechanisms underlying the unique vulnerability and/or resistance of particular subfields to a transient ischemic insult in the gerbil hippocampus. PMID- 9219966 TI - Chronic cerebral hypoperfusion by permanent internal carotid ligation produces learning impairment without brain damage in rats. AB - To investigate the influence of cerebral hypoperfusion on learning behaviours, we developed a novel rat cerebral hypoperfusion model, in which the bilateral internal carotid arteries were permanently ligated to reduce the cerebral blood flow, and examined its behavioural and histopathological consequences in comparison to those occurring after bilateral common carotid ligation. In the Morris water maze task, rats with common carotid ligation exhibited a learning deficit, whereas rats with internal carotid ligation exhibited normal learning. Both models exhibited significant learning impairments in the eight-arm radial maze task, although the impairment was less severe in internal carotid-ligated rats than in common carotid-ligated rats. The cerebral blood flow of rats with common carotid ligation was reduced significantly both two and 10 days after ligation, and was still below normal three months after ligation. A milder, but significant reduction in the cerebral blood flow was observed in internal carotid ligated rats. Shrinkage of the optic nerves and a circadian activity rhythm desynchronized to the light/dark cycle were exhibited by the rats with common carotid ligation, whereas these parameters remained unaffected in the rats with internal carotid ligation, suggesting that permanent ligation of common carotid arteries but not internal carotid arteries impairs visual functions. The main pathological changes observed in the brain following common carotid ligation were rarefaction and gliosis of the white matter and neuronal loss in the hippocampal CA1 region. On the other hand, the rats with internal carotid ligation had no significant brain damage. Chronic treatment with idebenone (1.5 and 15 mg/kg/day), a cerebral energy metabolism enhancer, over a three-month period, commencing five days after ligation, ameliorated the impairment of water maze learning in rats with common carotid ligation. The treatment also significantly improved the learning impairment in the radial maze task of internal carotid ligated rats. Idebenone had no effect on the histopathological changes that followed cerebral hypoperfusion. It is concluded that cerebral hypoperfusion induced by permanent internal carotid ligation impairs the working memory without causing pathological damage to the brain tissues and the visual system, and the learning impairment can be ameliorated by a cerebral energy metabolism enhancer. These findings have the clinical implication that a reduction in blood flow may be an important factor that causes or exacerbates cognitive decline in dementias. PMID- 9219967 TI - Cortical input to the basal forebrain. AB - The arborization pattern and postsynaptic targets of corticofugal axons in basal forebrain areas have been studied by the combination of anatomical tract-tracing and pre- and postembedding immunocytochemistry. The anterograde neuronal tracer Phaseolus vulgaris leucoagglutinin was iontophoretically delivered into different neocortical (frontal, parietal, occipital), allocortical (piriform) and mesocortical (insular, prefrontal) areas in rats. To identify the transmitter phenotype in pre- or postsynaptic elements, the tracer staining was combined with immunolabeling for either glutamate or GABA, or with immunolabeling for choline acetyltransferase or parvalbumin. Tracer injections into medial and ventral prefrontal areas gave rise to dense terminal arborizations in extended basal forebrain areas, particularly in the horizontal limb of the diagonal band and the region ventral to it. Terminals were also found to a lesser extent in the ventral part of the substantia innominata and in ventral pallidal areas adjoining ventral striatal territories. Similarly, labeled fibers from the piriform and insular cortices were found to reach lateral and ventral parts of the substantia innominata, where terminal varicosities were evident. In contrast, descending fibers from neocortical areas were smooth, devoid of terminal varicosities, and restricted to the myelinated fascicles of the internal capsule en route to more caudal targets. Ultrastructural studies obtained indicated that corticofugal axon terminals in the basal forebrain areas form synaptic contact primarily with dendritic spines or small dendritic branches (89%); the remaining axon terminals established synapses with dendritic shafts. All tracer labeled axon terminals were immunonegative for GABA, and in the cases investigated, were found to contain glutamate immunoreactivity. In material stained for the anterograde tracer and choline acetyltransferase, a total of 63 Phaseolus vulgaris leucoagglutinin varicosities closely associated with cholinergic profiles were selected for electron microscopic analysis. From this material, 37 varicosities were identified as establishing asymmetric synaptic contacts with neurons that were immunonegative for choline acetyltransferase, including spines and small dendrites (87%) or dendritic shafts (13%). Unequivocal evidence for synaptic interactions between tracer labeled terminals and cholinergic profiles could not be obtained in the remaining cases. From material stained for the anterograde tracer and parvalbumin, 40% of the labeled terminals investigated were found to establish synapses with parvalbumin-positive elements; these contacts were on dendritic shafts and were of the asymmetrical type. The present data suggest that corticofugal axons innervate forebrain neurons that are primarily inhibitory and non-cholinergic; local forebrain axonal arborizations of these cells may represent a mechanism by which prefrontal cortical areas control basal forebrain cholinergic neurons outside the traditional boundaries of pallidal areas. PMID- 9219968 TI - GABA(A) receptor-mediated inhibition in the nucleus ambiguus motoneuron. AB - GABA-sensitive ambiguus motoneurons were investigated by microiontophoretic application of GABAergic drugs including bicuculline and muscimol in alpha chloralose- and urethane-anaesthetized rats. Ambiguus motoneurons were activated by recurrent laryngeal nerve stimulation through a small cuff electrode and identified as laryngeal motoneurons when they met the conventional criteria for antidromic activation. GABA(A) antagonist, bicuculline, and its agonist, muscimol, were iontophoretized on ambiguus motoneurons through a three multibarrel electrode glued to the recording microelectrode. One-hundred and nineteen out of 155 neurons sampled from the loose formation and its vicinity were found to be respiratory neurons, most of which were inspiratory neurons. A small proportion (32 neurons) was classified as laryngeal motoneurons according to the criteria. A majority of laryngeal motoneurons was found to be GABA sensitive. Namely, application of GABA and its antagonist and agonist affected the antidromic spikes in a dose-dependent manner; GABA and muscimol usually decreased the amplitude and slowed the slope in the spike, whereas bicuculline, reversed these inhibitory effects. The dose-dependent relationships were limited exclusively to the measurements analysed in the negative-going phase but not in the positive-going phase in the antidromic spike. GABA and muscimol decreased but bicuculline increased the ratios to control in these measurements. The effects of distributions of the histograms shifting towards the opposite direction were statistically significant. The line of evidence suggests strongly that a majority of laryngeal motoneurons located in the nucleus ambiguus presumably possesses GABA(A) receptors on their postsynaptic membrane. These GABA-sensitive laryngeal motoneurons may receive inputs either from inhibitory interneurons subserving the reciprocal inhibition in the reflexive integration or from inhibitory respiratory interneurons which control the synchronized glottic movements during vocalization and respiration. PMID- 9219969 TI - Two types of cholinergic neurons in the rat tegmental pedunculopontine nucleus: electrophysiological and morphological characterization. AB - Two types of tegmental pedunculopontine nucleus neurons have been reported previously based on their electrophysiological characteristics: type I neurons were characterized by low-threshold Ca spikes and type II neurons displayed a transient outward current. This report describes the membrane properties, synaptic inputs, morphologies and axonal projections of two subgroups of type II neurons examined in an in vitro slice preparation. Type II neurons were divided into two groups based on their spike durations: short-duration neurons with an action potential duration of 0.7-1.5 ms and long-duration neurons with an action potential duration of 1.6- 2.9 ms. Choline acetyltransferase immunohistochemistry combined with biocytin labeling indicated that 56% of short-duration neurons and 61% of long-duration neurons were immunopositive for choline acetyltransferase. Short-duration neurons had a high input resistance and the capacity to discharge with high frequency. By contrast, long-duration neurons had a low input resistance and low firing frequency and upon current injection displayed an accommodation (spike-frequency adaptation) before reaching a steady firing frequency. Microstimulation of the substantia nigra pars compacta evoked antidromic responses in both short-duration neurons (n=5/14, 36%) and long duration neurons (n=20/39. 51%). Stimulations of the subthalamic nucleus and the substantia nigra pars reticulata induced in these neurons excitatory and inhibitory postsynaptic potentials, respectively. Short-duration neurons were dispersed equally throughout the extent of the tegmental pedunculopontine nucleus area, while long-duration neurons were located more in the rostral tegmental pedunculopontine nucleus. Short-duration neurons were small with two to four thin primary dendrites. Long-duration neurons were medium to large with three to six thick primary dendrites. Cell size was positively correlated with spike duration and axonal conduction velocity, but negatively with input resistance and spontaneous firing frequency. Both groups of neurons had ascending (toward thalamus, pretectal areas and tectum) and descending (toward pontomedullary reticular formation) axons in addition to nigropetal axons. Ascending axons were observed in 75% (6/8) of short-duration neurons and in 45% (15/33) of long duration neurons, while nigropetal axons were observed in 50% (4/8) of short duration neurons and in 76% (25/33) of long-duration neurons. These results suggest that the tegmental pedunculopontine nucleus cholinergic projection system is composed of heterogeneous populations of neurons in terms of electrophysiological and morphological characteristics as well as their distribution patterns in the nucleus. PMID- 9219970 TI - Distribution of the messenger RNA for the extracellularly regulated kinases 1, 2 and 3 in rat brain: effects of excitotoxic hippocampal lesions. AB - Neurofibrillary tangles in Alzheimer's disease are composed of hyperphosphorylated forms of the microtubule-associated protein tau. Based on biochemical criteria, several enzymes have emerged as potential tau protein kinases, including the extracellularly regulated kinases 1, 2 and 3. In situ hybridization was used to map the messenger RNA distribution of extracellularly regulated kinase 1, 2 and 3 in the adult rat brain and their response to excitotoxic hippocampal lesions was examined. Extracellularly regulated kinase 1 messenger RNA was uniformly expressed by glia, but was also present in the dentate gyrus and some other neuronal populations. Extracellularly regulated kinase 2 was exclusively neuronal and concentrated within the cortical laminae and the CA subfields of the hippocampal formation. Extracellularly regulated kinase 3 messenger RNA expression was similar to extracellularly regulated kinase 2 and was also present in neurons but the level of expression was lower. Extracellularly regulated kinases 2 and 3 messenger RNA expression was lost following excitotoxic injury, further supporting a neuronal localization. Extracellularly regulated kinase 1 messenger RNA expression appeared unaltered, suggesting a non-neuronal localization and lack of responsiveness to lesion at the level of transcription. By contrast, messenger RNA of sgk, a recently described serine/threonine kinase, was up-regulated by glial cells following excitotoxic injury. Based on their messenger RNA distribution, cellular localization and response to lesion, it is clear that each kinase may function differently in various signaling pathways. Extracellularly regulated kinase 2, however, is the only kinase with the proper messenger RNA distribution to contribute to neurofibrillary tangle formation in Alzheimer's disease. PMID- 9219971 TI - The distribution of the GABA(A) beta2,beta3 subunit receptor in the cat superior colliculus using antibody immunocytochemistry. AB - GABA-containing synaptic terminals in the cat superior colliculus include two varieties of presynaptic dendrite and at least one type of axon terminal with flattened vesicles. These anatomically distinct synaptic profiles probably also mediate different types of inhibition. Whether they are associated with different types of GABA receptor is unknown and one objective of the present paper. We used the antibody mAb 62-361 directed against the beta2,beta3 subunits of the GABA(A) receptor complex to determine whether the distribution of this receptor subunit is specific to one or more types of GABA-containing synapse. At the light microscope level, beta2,beta3 immunoreactivity was densely distributed within the neuropil of the zonal and superficial gray layers, and more lightly within the optic, intermediate, and deep gray layers. No cell bodies were labelled by the antibody in the zonal and superficial gray layers, but numerous cells contained internalized cytoplasmic immunoreactivity in the optic, intermediate gray, and deeper layers. At the ultrastructural level, synaptic sites opposite axon terminals that contained flattened synaptic vesicles (F profiles) were often beta2,beta3 immunoreactive, while postsynaptic sites opposite presynaptic dendrites (PSD profiles) were never immunoreactive. The label at F profiles usually filled the synaptic cleft and coated the postsynaptic plasma membrane. Some membrane-associated label was also found at non-synaptic sites. We conclude that this receptor subunit is selectively associated with flattened vesicle axon terminals and not with presynaptic dendrites, a result which supports evidence that those terminal types mediate different types of inhibition. PMID- 9219972 TI - Discrete cellular and subcellular localization of glutamine synthetase and the glutamate transporter GLAST in the rat vestibular end organ. AB - Glial cells play an important role in the removal and metabolism of synaptically released glutamate in the central nervous system (CNS). It is not clear how glutamate is handled at peripheral glutamate synapses, which are not associated with glia. Glutamate is a likely transmitter in the synapse between the hair cells and afferent dendrites of the vestibular end organ. Immunocytochemistry was performed to investigate the distribution at this site of the high affinity glutamate transporter GLAST and glutamate metabolizing enzyme glutamine synthetase. Confocal microscopy revealed that GLAST and glutamine synthetase were co-localized in supporting cells apposed to the immunonegative hair cells. Postembedding immunoelectron microscopy revealed that GLAST was heterogeneously distributed along the plasma membranes of the supporting cells, with higher concentrations basally (at the level of the afferent synapses) than apically. Both immunoreactivities were also present in non-neuronal cells in the vestibular ganglion. The present findings suggest that glutamate released at the afferent synapse of vestibular hair cells may be taken up by adjacent supporting cells and converted into glutamine. Thus, at this peripheral synapse, the supporting cells may carry out functions similar to those of glial cells in the CNS. PMID- 9219973 TI - Plasmalemmal ATPase calcium pump localizes to inner and outer hair bundles. AB - Recent studies demonstrate calcium ion influx at the tips of hair cell stereocilia during mechano-transduction. These ions must be either pumped from the cytosol into the extracellular space or endoplasmic envelope, or else sequestered by binding to specific proteins. A plasma membrane calcium pump (ATPase-type) was analysed in whole-mounts of rat organ of Corti using a monoclonal antibody to a large cytoplasmic loop of this protein. The reactivity was particularly high on the tips of longer stereocilia and was found along the shafts. Inner hair cell stereocilia had much less reactivity than outer hair cells. The reactivity lined the plasma membrane of inner hair cell bodies while a higher reactivity appeared in the cytoplasm of outer hair cells. Supporting cells were unreactive. Ultrastructural examination confirmed the plasma membrane calcium pump location on stereocilia and along the endolymph surface of receptor cells. Reaction product lined the plasma membrane of stereocilia as intense puncta. More reactive puncta occurred near the distal ends of stereocilia and the number decreased toward the ciliary base. The endolymph plasma membrane over the cuticular notch was especially reactive. The finding of more intense pump reactivity at the tips of stereocilia than the base is consistent with the hypothesis that during transduction, calcium ions enter stereocilia, distally, and the ATPase plasma membrane calcium pump rapidly extrudes these ions to the extracellular space. PMID- 9219974 TI - Localization of estrogen receptor protein and estrogen receptor messenger RNA in peripheral autonomic and sensory neurons. AB - The presence of estrogen receptor protein and estrogen receptor messenger RNA was revealed in peripheral ganglionic neurons of the rat. The pelvic parasympathetic autonomic ganglion and lumbosacral dorsal root sensory ganglia were examined for estrogen receptor-containing neurons because they have known projections to the uterus and uterine cervix. The vagal nodose ganglia were studied for estrogen receptor-containing neurons because they are suspected sources of influence on the uterus. Immunohistochemistry. in situ hybridization histochemistry and retrograde tracing were utilized. Immunoreactivity for estrogen receptors was evident in the nuclei of a subpopulation of neurons in the pelvic ganglia, sixth lumbar and first sacral dorsal root ganglia and nodose ganglia. Some estrogen receptor-positive neurons also contained the retrograde tracer FluoroGold that previously had been injected into the uterus and uterine cervix. Estrogen receptor messenger RNA was also evident in a subpopulation of ganglionic neurons. These data suggest that a certain population of neurons in autonomic and sensory ganglia are capable of synthesizing estrogen receptors and these receptors can serve as binding sites for estrogen. Thus, certain aspects of the structure, function and neurochemistry of some autonomic and sensory neurons may be influenced by the sex steroid estrogen. PMID- 9219975 TI - Central c-Fos expression in neonatal and adult rats after subcutaneous injection of hypertonic saline. AB - Centrally-mediated responses to plasma hyperosmolality include compensatory drinking and pituitary secretion of vasopressin and oxytocin in both adult and neonatal rats. However, the anorexia that is produced by plasma hyperosmolality in adult rats is not evident in neonates, perhaps due to functional immaturity of osmoresponsive hindbrain circuits. To examine this possibility, the present study compared treatment-induced brain expression of the immediate-early gene product c Fos as a marker of neural activation in adult and two-day-old rats after subcutaneous injection of 2 M NaCl (0.1 ml/10 g body weight). This treatment produced marked hypernatremia in adult and two-day-old rats without altering plasma volume. Several brain regions (including components of the lamina terminalis, the paraventricular and supraoptic nuclei of the hypothalamus, and the area postrema) were activated to express c-Fos similarly in adult and two-day old rats after 2 M NaCl injection, consistent with previous reports implicating a subset of these regions in osmotically-stimulated drinking and neurohypophyseal secretion. In contrast, other areas of the brain that were activated to express c Fos in adult rats after 2 M NaCl injection were not activated in neonates: these areas included the central nucleus of the amygdala, the parabrachial nucleus and catecholamine cell groups within the caudal medulla. This study demonstrates that certain brain regions that are osmoresponsive in adult rats (as defined by induced c-Fos expression) are not osmoresponsive in two-day-old rats. When considered in the context of known differences between the osmoregulatory capacities of adult and neonatal rats, our results are consistent with the idea that osmoresponsive forebrain centres are primarily involved in osmotically stimulated compensatory drinking and neurohypophyseal secretion, whereas osmoresponsive regions of the hindbrain are important for concomitant inhibition of feeding and gastric emptying. PMID- 9219976 TI - Role of neutrophils in spinal cord injury in the rat. AB - Activated neutrophils are thought to be involved in tissue injury through the release of various inflammatory mediators. To understand the role of neutrophils in spinal cord injury, the effects of nitrogen mustard-induced leukocyte depletion and the administration of an anti-P-selectin monoclonal antibody on motor disturbances observed following spinal cord compression were examined in rats. Spinal cord injury was induced by applying a 20-g weight for 20 min at the level of the 12th thoracic vertebra, resulting in motor disturbances of the hindlimbs 24 h postcompression. Motor disturbances, evaluated using Tarlov's index, an inclined-plane test and climbing ability, were markedly attenuated in rats with nitrogen mustard-induced leukocytopenia. Administration of the anti-P selectin monoclonal antibody, by which adhesion of activated neutrophils to endothelial cells may be inhibited, also attenuated motor disturbances. Histological examination revealed that intramedullary hemorrhages observed 24 h after compression at the 12th thoracic vertebra of the spinal cord were significantly attenuated in leukocytopenic animals and those which received the anti-P-selectin monoclonal antibody. The accumulation of neutrophils at the site of compression, as evaluated by measuring the tissue myeloperoxidase activity, significantly increased with time following the compression, peaking at 3 h postcompression. Spinal cord myeloperoxidase activity did not increase in sham operated animals. Leukocyte depletion and administration of the anti-P-selectin monoclonal antibody both reduced the accumulation of neutrophils in the damaged spinal cord segment 3 h postcompression. These observations strongly suggest that activated neutrophils play an important role in compression-induced thoracic spinal cord injury and that a P-selectin-mediated interaction between activated neutrophils and endothelial cells may be a critical step in endothelial cell injury leading to spinal cord injury. PMID- 9219977 TI - Differential expression of galanin immunoreactivities in the primary sensory neurons following partial and complete sciatic nerve injuries. AB - Neuropeptide expression in primary sensory neurons is highly plastic in response to peripheral nerve axotomy. While neuropeptide changes following complete sciatic nerve injury have been extensively studied, much less is known about the effects of partial sciatic nerve injuries on neuropeptide plasticity. Galanin. a possible endogenous analgesic peptide, was up-regulated in primary sensory neurons following complete sciatic nerve injury. We investigated the effects of partial sciatic nerve injuries on galanin expression in primary sensory neurons, and compared this effect with that after complete sciatic nerve injury. Complete transection, partial transection and chronic constriction injury were made, respectively, on the sciatic nerves of three groups of rats at high thigh level. Animals were allowed to survive for four and 14 days before being killed. L4 and L5 dorsal root ganglia, L4 5 spinal cord and lower brainstem were processed for galanin immunocytochemical staining. After all three types of sciatic nerve injuries, galanin-immunoreactive neurons were significantly increased in the ipsilateral dorsal root ganglia, and galanin-immunoreactive axonal fibres were dramatically increased in the superficial laminae of the dorsal horn and the gracile nuclei, compared to the contralateral side. However, in partial injury models, the percentages of galanin-immunoreactive dorsal root ganglion neurons were significantly higher than in complete nerve transection. Size frequency distribution analysis detected that more medium- and large-size galanin immunoreactive dorsal root ganglion neurons were present after partial nerve transection and constriction injury than after complete nerve transection. Using a combined approach of retrograde tracing of flurorescent dyes and galanin immunostaining, we found that a partial transection increased the proportions of galanin-immunoreactive neurons among both axotomized and non-axotomized neurons. Galanin-immunoreactive axonal fibres were not only detected in the superficial laminae, but also in the deeper laminae of the dorsal horn of partial injury animals. Furthermore, more galanin-immunoreactive axonal fibres were observed in the ipsilateral gracile nuclei of partially injured rats than in completely injured rats. We conclude that partial sciatic nerve injuries induced greater galanin up-regulation in medium- and large-size dorsal root ganglion neurons than complete sciatic nerve injury. Galanin expression in primary sensory neurons seems to be differentially regulated following partial and complete sciatic nerve injuries. PMID- 9219978 TI - Differential regulation of substance P by all members of the nerve growth factor family of neurotrophins in avian dorsal root ganglia throughout development. AB - This study examined the effects of nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3 and neurotrophin-4/5 on substance P levels in dorsal root ganglia of the quail shortly after ganglia formation (stage 26, embryonic day 4.5), during the middle of development (stage 33, embryonic day 7.5) and during late development (stage 44, embryonic day 14). It has already been shown that nerve growth factor increases levels of substance P during the middle and late stages of development, and that messenger RNA for the neurotrophin receptors, trkA, trkB and trkC is present at all of these stages. Dorsal root ganglia were isolated, rinsed with defined medium to dilute endogenous neurotrophins and exposed to one of the neurotrophins for either 4 or 20 h. Substance P levels were quantitated using enzyme immunoassay. None of the neurotrophins had any effect on substance P levels in dorsal root ganglia obtained at stage 26 after either a 4 or 20 h exposure time. Nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3 and neurotrophin-4/5 all significantly increased levels of substance P after either a 4 h or 20 h incubation in ganglia obtained at stages 33 and 44. The effects of nerve growth factor and neurotrophin-3 were specific: increases in substance P were completely blocked by simultaneous exposure to antibodies against either nerve growth factor or neurotrophin-3. The absence of any effect of neurotrophins on substance P expression during early development was unexpected, since dorsal root ganglia exhibit substantial levels of substance P and receptors for the neurotrophins are present and are apparently functional. It was also surprising that brain-derived neurotrophic factor, neurotrophin-3 and neurotrophin-4/5 induced increases in substance P levels during the middle and late stages of development, since substance P was thought to be exclusively localized to small TrkA neurons in dorsal root ganglia. However, immunocytochemical examination of dorsal root ganglia at stages 33 and 44 revealed substance P-like immunoreactivity in larger neurons as well as in small neurons. The results of this study have shown that different cellular responses to neurotrophins, such as effects on survival and/or peptide expression, may be acquired with differing temporal patterns not strictly related to expression of their receptors. Further, the regulation of neuropeptide synthesis in dorsal root ganglia is not due to any one neurotrophic factor. and the factors that regulate expression during early development are still unknown. PMID- 9219979 TI - Ultrastructural co-localization of calmodulin and B-50/growth-associated protein 43 at the plasma membrane of proximal unmyelinated axon shafts studied in the model of the regenerating rat sciatic nerve. AB - Calmodulin and de-phosphorylated B-50/growth-associated protein-43 (GAP-43) have been shown to bind in vitro in a molecular complex, but evidence for an in situ association in the nervous system does not exist. Previously, we have reported that, in the model of the regenerating rat sciatic nerve, the B-50/GAP-43 immunoreactivity is increased and concentrated at the axolemma of unmyelinated axons located proximal to the site of injury and axon outgrowth. To explore a putative function of B-50/GAP-43, namely, the capacity of binding calmodulin to the plasma membrane, we examined the ultrastructural distribution of calmodulin in the proximal unmyelinated axon shafts of this model, using double immunolabelling and detection by fluorescent or gold probes conjugated to second antibodies. Immunofluorescence showed that seven days post-sciatic nerve crush the calmodulin immunoreactivity, similar to B-50/GAP-43 immunoreactivity, was intense in unmyelinated axon shafts located proximal to the site of injury of the regenerating nerve. Ultrastructurally, calmodulin was located at the axolemma of these regenerating unmyelinated axon shafts and inside the axoplasm, where it was associated with vesicles and microtubules. The plasma membrane labelling (approximately 69%) was significantly higher than the axoplasmic labelling. Over 60% of the plasma membrane-associated calmodulin co-localized with B-50/GAP-43 in a non-random distribution. Since normally calmodulin is largely present in the cytoplasm, these data suggest that calmodulin has been concentrated at the plasma membrane of unmyelinated axons, most probably by B-50/GAP-43. If the concentrating effect is due to B-50/GAP-43, then there is a possibility that these proteins may be present as a molecular complex in situ. The physiological significance could be that this association regulates the local availability of both B-50/GAP-43 and calmodulin for other interactions. PMID- 9219980 TI - Molecular characterization and functional expression of a substance P receptor from the sympathetic ganglion of Rana catesbeiana. AB - Substance P is an important neuropeptide neurotransmitter in the central, autonomic and enteric nervous systems. In sympathetic ganglia, substance P is thought to play a role in modulating synaptic transmission. Release of substance P by neuronal stimulation or direct application of substance P to ganglionic neurons increases neuronal excitability. An amphibian substance P receptor complementary DNA has been cloned and characterized from bullfrog, Rana catesbeiana, sympathetic ganglion complementary DNA libraries. The deduced primary structure contains features indicative of a seven transmembrane domain G protein-coupled receptor. The deduced protein sequence shows 69% identity to previously cloned mammalian substance P receptors. In situ hybridization analysis performed on bullfrog sympathetic ganglia using digoxigenin-labelled complementary RNA probe demonstrated that approximately 75% of the principal neurons displayed reaction product above background levels. Radioligand binding studies were performed on stably transfected cells with [(125)I]Tyr-1-substance P as the ligand. Substance P had an IC50 of 16 nM and the agonist potency profile was substance P>neurokinin A >> neurokinin B. The order of potency for three tachykinins to increase intracellular calcium when applied to a stably transfected clonal cell line was substance P>neurokinin A >> neurokinin B. This order of agonist potency also held for inhibition of the M-type potassium current in intact bullfrog sympathetic neurons. The non-peptide substance P antagonists CP-96345 and RP-67580 at concentrations that block mammalian substance P receptors had little or no effect on the responses to substance P at the bullfrog receptor. Overall, these results demonstrate that the cloned sequence has the features consistent with and characteristic of a substance P receptor. The results are discussed with reference to the established pharmacology of the bullfrog substance P receptor and known structure activity relationships of mammalian tachykinin receptors. PMID- 9219981 TI - Electrophysiological demonstration of N-methyl-D-aspartate receptors at the afferent synapse of catfish electroreceptor organs. AB - An excitatory amino acid, most probably L-glutamate, acts as a neurotransmitter at the receptor cell--afferent fibre synapses in the ampullary electroreceptor organs of the freshwater catfish Ictalurus nebulosus. In the present study, we have used an electrophysiological approach to investigate the presence of N methyl-D-aspartate receptors at this level. N-Methyl-D-aspartate, dissolved in an Mg(2+)-containing (normal) solution, had no effect on afferent activity, not even at 5 mM. However, addition of 5 mM N-methyl-D-aspartate to an Mg(2+)-free solution evoked an enduring increase in firing rate. The application of N-methyl D-aspartate combined with electrical sine wave stimulation produced a firing increase in the primary afferents, even in the presence of Mg2+ (1.5 mM). Glycine (0.01-0.001 mM) significantly potentiated the N-methyl-D-aspartate responses. Addition of antagonists of the actions of N-methyl-D-aspartate, 7-chlorokynurenic acid, DL-2-amino-5-phosphonovaleric acid and ketamine in concentrations of 0.5 2.0 mM led to a decrease in resting and stimulus-evoked activity. 7 Chlorokynurenic acid also blocked the responses to application of N-methyl-D aspartate. The glycine agonist D-serine (0.01 mM) prevented the 7-chlorokynurenic inhibitory effect. These results suggest the involvement of N-methyl-D-aspartate receptors in mediating the actions of L-glutamate at the afferent synapses of the electroreceptor organs of the catfish. PMID- 9219982 TI - Re-examination of incisal tooth wear in children and adolescents. AB - The material presented in this report was derived from a longitudinal study of the development and progress of incisal tooth wear in children and adolescents. The study group was established in 1991-1992 (baseline examination) and consisted of participants between 8 and 15 years of age. They were re-examined in 1993 (interim examination) and in 1994 (final examination). In all, 77 school children (30 females and 47 males) participated in the three examinations. The timespan between baseline and interim was on average 15 months and the period between interim and final examination averaged 16 months. The total observation period was approximately 32 months. Assessments of incisal wear was made on stone casts using the incisal wear index which was shown to have good reproducibility. The results demonstrated that the prevalence and severity of incisal wear had increased significantly with age. In general the rate of incisal wear progression was higher between baseline and interim than between interim and final examination. In this respect certain differences between the tooth types seemed to exist. At baseline the relationship between age and incisal wear was positive and statistically significant for all tooth types. At the final examination the strength of this relationship had decreased for maxillary and mandibular central and lateral incisors. For canine teeth of both jaws the relationship between age and incisal wear was no longer statistically significant at the final examination. PMID- 9219983 TI - Relationship between incisal tooth wear and the increasing number of permanent teeth in children and adolescents. AB - The material presented in this report was derived from a longitudinal study of the development and progress of incisal tooth wear in children and adolescents. The study group was established in 1991/1992 and consisted of 77 participants between 8 and 15 years old. All participants were re-examined in 1994. The interval between the first and second examination was approximately 32 months. Each participant was scored for tooth wear of the anterior teeth of both jaws according to the Incisal wear Index (IwI) and the number of permanent teeth and the tooth types present were recorded at each examination. The results showed that the pattern of incisal wear had been maintained during the observation period. Severity of incisal wear increased as the number of teeth increased. The strength of the relationship between incisal wear and the number of teeth decreased during the observation period. At the same time, the number of teeth and the size of the wear increments were negatively correlated indicating that the rate of wear progression decreased as the number of teeth increased. PMID- 9219984 TI - Titanium for removable dentures. II. Two-year clinical observations. AB - Titanium (Ti) is a relatively new metal in prosthodontics. As a possible material for removable partial dentures (RPDs), it was used in its pure form to make 10 RPDs which were compared with 10 identical cobalt-chromium (Co-Cr) alloy RPDs in a clinical trial. The dentures were used alternately for four weeks, the subjects were asked to answer a questionnaire and then the Co-Cr dentures were withdrawn. The Ti dentures were then followed up for 2 years and another standardized questionnaire administered. A survival rate of rests and retainers of 91% was recorded and the dentures were found to be more comfortable (55%) and preferred (64%) than the Co-Cr dentures. Factors to increase the success rates are discussed. PMID- 9219985 TI - Effect of complete denture renewal on oral health--a survey of 42 patients. AB - The effect of complete denture renewal on oral health was evaluated both subjectively and clinically at follow-up 30 months (range 19-36 months) after completion of treatment in 42 edentulous patients (31 women and 11 men, aged 34 76 years) treated by dental students during 1989-1992. Ninety per cent of the patients were satisfied with the new appearance of their dentures and 71% with the way they functioned. When comparing the base data and results from follow-up it seems that general health and medication, anatomical circumstances, salivary flow rates and denture wearing habits in edentulous subjects do not change significantly over a few years. The main effects of denture renewal are seen in patient satisfaction, and clinically in the improved condition of oral mucosa and better fit and acceptable occlusion of dentures. PMID- 9219986 TI - Analysis of two methods for occlusal contact registration with the T-Scan system. AB - Various authors have studied the reproducibility of occlusal contact by means of the T-Scan computerized system and obtained contradictory results. In the present work the value of the T-Scan system as a method for exploring occlusion has been analysed. For this purpose, the same variable, i.e. the number of contacts, was recorded in 31 subjects using the two operation modes enabled by the T-Scan and the respective results were then compared. After an analysis of variance was performed to test the equality of averages of the number of tooth contacts for each patient in the four positions studied (maximum intercuspation, edge to edge protrusion, right laterality and left laterality) in the time and force analysis modes, the results obtained showed that the number of occlusal contacts is significantly different for each patient both in the various mandibular positions and in the force and time analysis modes, being proportionally greater in the latter case. PMID- 9219987 TI - Effect of cleaning dentine with soap and pumice on shear bond strength of dentine bonding agents. AB - This in vitro study reports on the cleaning effect of different soaps on the shear bond strength of various dentine-bonding agents. Human teeth were coated with provisional cements for 24 h or for 14 days. After removing the provisional cements with a scaler, the dentinal surface was cleaned with a cotton pellet and non-fluoridated flour of pumice and soap for 10 sec. Different dentine-bonding agents and a luting resin were bonded to the dentinal surface according to manufacturers' instructions with the bonding agent and the composite material being light-cured at the same time. The bonding agents were tested under intrapulpal pressure and with thermal cycling to imitate physiological conditions. Compared with cleaning the dentine with water and pumice, all soaps investigated in this study decreased the shear bond strength values of the tested dentine-bonding agents considerably. PMID- 9219988 TI - Electromyographic evidence for a digastric reflex evoked by perioral stimuli in humans. AB - Electromyographic recordings (EMGs) were made using skin surface electrodes placed over the anterior digastric muscle in seven subjects. In every case, short bursts of electrical stimuli to the upper lip produced a response in the EMG that had a minimum latency of 62.0 +/- 10.8 ms (mean +/- SD). By contrast, no responses were seen when single pulse stimuli were applied. In 6/7 subjects, the minimum stimulation intensity that produced the reflex was described as being sharp or painful. In three additional experiments, single motor units were recorded within the digastric muscle using needle electrodes. In two of these experiments, there was evidence of reflex activity 60-110 ms after the application of painful electrical stimuli to the lip. These findings confirm that perioral stimuli can evoke a digastric reflex in humans and suggest that this reflex requires the summation that results from successive volleys of impulses in a large number of nociceptive afferent neurones. PMID- 9219989 TI - Brazing joints of gold alloy used in porcelain-fused-to-metal restorations and their resistance to deflection fatigue. AB - The aim of this study was to compare the resistance to deflection fatigue of a gold alloy used in porcelain-fused-to-metal restorations with and without a brazing joint. Pre-ceramic brazing filler metal was used to join the parent specimens of gold alloy together. The deflection fatigue test was carried out mainly with 0.4 m deflection of the test specimens (n = 5) but to obtain an S-N curve for the specimens, other magnitudes of deflection, i.e. the stress, were also used. When the fracture surface of the test specimens was examined by scanning electron microscopy (SEM), the results showed that the brazing joint in the gold alloy test specimen decreases the fatigue resistance considerably compared to that of specimens without a brazing joint (P = 0.002). SEM examination showed that the failure type of the brazing joint was cohesive and that the brazing filler metal had a more porous structure than the parent gold alloy. These results suggest that, due to the occlusal biting forces in situ, the brazing joints in fixed partial dentures can be fractured by metal fatigue. PMID- 9219990 TI - Changes of surface texture of enamel in vivo. AB - There are various methods of measuring tooth wear, including that which could be considered to be associated with acid erosion. However, the present systems are limited in that they require an extended period of study to detect wear and also they do not give any insight as to the underlying mechanism of the tooth tissue loss. The aim of this study was to use a profilometric measuring system analysing novel surface texture parameters normally associated with the automobile industry to describe the wear and run-in behaviour of machined sliding engine parts. The texture parameters selected were the traditional Ra parameter and also parameters derived from the bearing ratio. Analyses of the surface texture of the labial surface of upper central incisors at an interval of three months indicated that the enamel surface was smoother but there were subtle changes occurring to the enamel. There was a statistically significant increase in the depth of pits/pores of the enamel surface, which could be identified as sites of retention of exogenous acid or chelating agent. The effects of acid erosion are not uniform but are dependent upon several factors; the configuration of the enamel surface may play an important part in the mediation of acid erosion type lesions. PMID- 9219991 TI - Unerupted canine without median diastema. AB - The eruption of the canine is known to be a factor for diastema closure. In this study the relative changes in the orientation of the unerupted canine were assessed using the orthopantomographs of 9854 patients who sought consultation between April 1984 and March 1993. A total of 38 canines in 32 patients, all aged 11 years or older, were identified as unerupted canines. The features of the patients with unerupted canine showed no significant relation to diastema closure status, but some patients had unerupted horizontal or inverted canine without diastema even in the absence of a history of orthodontic treatment, suggesting the presence of a mechanical force due to some phenomenon other than canine eruption as a factor in diastema closure. PMID- 9219992 TI - Chewing activity and activities of daily living in the elderly. AB - The present study was conducted in order to determine the statistical relationship between chewing activity and activities of daily living (ADL) in the elderly. Subjects which took part were 79 elderly individuals (37 males, 42 females) ranging in age from 65 to 74 years. Based on questionnaires regarding diet, a mastication score was determined to evaluate chewing activity in each subject. This score was scaled from 0 to 100%. ADL levels were determined using two indices: the guidelines of the Ministry of Health and Welfare of Japan and the Tokyo Metropolitan Institute of Gerontology (TMIG) index. The TMIG index is a multidimensional 13-item index of competence in which scores range from 0 to 13 points. According to the guidelines of the Ministry of Health and Welfare, the independence level of each subject was evaluated as independent, home-bound, or bedridden. These independence levels showed a significant correlation with the mastication score (P < 0.01). The mastication score also significantly correlated with the TMIG index, showing a Spearman coefficient of correlation of 0.63 in males (P < 0.01) and 0.71 in females (P < 0.01). These results suggest that chewing activity is related to ADL levels in elderly subjects. PMID- 9219993 TI - Oral awareness and ability to detect dental plaque. AB - Individuals exist who are unaware of the plaque on their teeth and unable to feel with their tongue any difference between tooth surfaces that are smooth and plaque free and those that are irregular or rougher because of a plaque coating. Oral form recognition (OFR), oral roughness discrimination (ORD) and plaque detection ability were examined in 46 undergraduate dental students. Forty-five of the students also completed Form C of Cattell's 16 PF Questionnaire. No relationship was found between the subjects' ability to detect dental plaque and their skills at either distinguishing between different surface roughnesses or recognizing forms orally. The association between personality and oral awareness was unclear. PMID- 9219994 TI - Denture stomatitis in an elderly edentulous Asian population. AB - Denture stomatitis is a common oral disease in denture wearers. Multiple aetiological and predisposing factors are believed to be responsible for its initiation and progression. The aim of this study was to assess the relationship between denture age, denture hygiene habits, denture wearing and denture cleanliness in an elderly edentulous Asian population. Seventy-five edentulous patients, all wearing maxillary complete dentures were divided into two groups. The test group comprised 36 patients (14 male and 22 female) with Type II denture stomatitis. The control group comprised 39 subjects (14 male and 25 female) with clinically healthy palatal mucosa. A standardized interview and clinical appraisal were carried out. The dye disclosing method was used to assess denture cleanliness and the resultant staining pattern scored. Statistical appraisal between the two groups revealed significant differences in denture hygiene habits (P < 0.05), denture wearing behaviour (P < 0.01) and denture cleanliness (P < 0.01). No significant difference was observed in the age of dentures between the test group and controls (P > 0.05). In the studied Asian edentulous population, a relationship between denture hygiene habits, denture wearing behaviour and denture cleanliness to the presence of denture stomatitis was observed. PMID- 9219995 TI - Comparison between the sequential and simultaneous approaches for the restoration of adjacent proximal cavities. AB - The clinical situation in which several proximal carious lesions have to be treated is frequently encountered. The need to provide a cost-effective and time effective treatment often compels the clinician to perform multiple adjacent proximal restorations simultaneously, at the same treatment session. Alternatively, such treatments may be performed sequentially in successive sessions. The different possible treatment modalities and their clinical sequel are described. Examination of the clinical results suggests that the preferred restorative procedure is the sequential one, enabling an optimal restoration of the original anatomical form, which is important for the proper health and function of the teeth and the surrounding periodontal tissues. PMID- 9219996 TI - Reinforced glass-ionomer cements: the influence of conditioners on marginal leakage. AB - The purpose of this in vitro study was to evaluate the influence of conditioners on the enamel and dentine margin sealing ability of three different reinforced glass-ionomer cements. Two Class V preparations were made on the buccal and lingual surfaces of 36 freshly extracted molar teeth. Preparations were solely in enamel or dentine/cementum. The teeth were randomly divided into three groups of 12 and restored with either Ketac Silver (KS), Hi-Dense (HD) or Miracle-Mix (MM) with and without (-C) their respective conditioners. All materials were capsulated and were manipulated according to the manufacturers' instructions. The restorations were finished as recommended by the manufacturers and then stored in saline at 37 degrees C for 1 week, polished, thermally stressed, subjected to dye penetration, sectioned and scored. Rankings in the order of decreasing leakage were as follows: enamel margin KS > KS-C > HD-C > HD > MM > MM-C; dentine margin KS > HD-C > KS-C > HD > MM-C > MM. At the enamel margins, only HD showed a significant increase in leakage when conditioner was not used. At the dentine margin, however, KS had significantly more leakage than KS-C and HD-C had significantly more leakage than HD. There was no significant difference in leakage for MM both with and without conditioner. The influence of conditioners on marginal leakage appears to be both product and tissue specific. PMID- 9219997 TI - Regulation of directed motility in Myxococcus xanthus. AB - Myxococcus xanthus is a Gram-negative bacterium that exhibits a complex life cycle. During vegetative growth, cells move as large swarms. However, when starved, cells aggregate into fruiting bodies and sporulate. Both vegetative swarming and developmental aggregation require gliding motility, which involves the slow movement of cells on a solid surface in the absence of flagella. The frequency of cell reversals controls the direction of movement and is regulated by the frz genes, which encode the 'frizzy' signal-transduction proteins. These proteins contain domains which bear striking similarities to the major chemotaxis proteins of the enteric bacteria: CheA, CheY, CheW, CheR, CheB and Tar. However, significant differences exist between the Myxococcus Frz proteins and the enteric Che/MCP proteins. For example, the Frz system contains three CheY-like response regulator domains: one is present on FrzE, which also contains a CheA-like domain, and two are present on FrzZ, which is a novel protein required for attractant, but not for repellent, responses. The identification of multiple CheY homologues in this system indicates a more complex regulatory pathway than that found in the enteric bacteria. While responses to repellent stimuli appear to follow the enteric paradigm, responses to attractants during vegetative swarming and development are more complex and may involve self-generated autoattractants. The Frz signal-transduction system regulates directed motility in M. xanthus and is essential for controlling both fruiting-body development and vegetative swarming. PMID- 9219998 TI - Quorum sensing by peptide pheromones and two-component signal-transduction systems in Gram-positive bacteria. AB - Cell-density-dependent gene expression appears to be widely spread in bacteria. This quorum-sensing phenomenon has been well established in Gram-negative bacteria, where N-acyl homoserine lactones are the diffusible communication molecules that modulate cell-density-dependent phenotypes. Similarly, a variety of processes are known to be regulated in a cell-density- or growth-phase dependent manner in Gram-positive bacteria. Examples of such quorum-sensing modes in Gram-positive bacteria are the development of genetic competence in Bacillus subtilis and Streptococcus pneumoniae, the virulence response in Staphylococcus aureus, and the production of antimicrobial peptides by several species of Gram positive bacteria including lactic acid bacteria. Cell-density-dependent regulatory modes in these systems appear to follow a common theme, in which the signal molecule is a post-translationally processed peptide that is secreted by a dedicated ATP-binding-cassette exporter. This secreted peptide pheromone functions as the input signal for a specific sensor component of a two-component signal-transduction system. Moreover, genetic linkage of the common elements involved results in autoregulation of peptide-pheromone production. PMID- 9219999 TI - The Bacillus subtilis DivIVA protein targets to the division septum and controls the site specificity of cell division. AB - The Bacillus subtilis divIVA gene, first defined by a mutation giving rise to anucleate minicells, has been cloned and characterized. Depletion of DivIVA leads to inhibition of the initiation of cell division. The residual divisions that do occur are abnormally placed and sometimes misorientated relative to the long axis of the cell. The DivIVA phenotype can be suppressed by disruption of the MinCD division inhibitor, suggesting that DivIVA controls the topological specificity of MinCD action and thus septum positioning. A DivIVA-GFP fusion targets to new and used sites of cell division, consistent with it having a direct role in topological specification. PMID- 9220000 TI - Regulation, replication, and integration functions of the Vibrio cholerae CTXphi are encoded by region RS2. AB - CTXphi is a filamentous phage that encodes cholera toxin, one of the principal virulence factors of Vibrio cholerae. CTXphi is unusual among filamentous phages because it can either replicate as a plasmid or integrate into the V. cholerae chromosome at a specific site. The CTXphi genome has two regions, the 'core' and RS2. Integrated CTXphi is frequently flanked by an element known as RS1 which is related to RS2. The nucleotide sequences of RS2 and RS1 were determined. These related elements contain three nearly identical open reading frames (ORFs), which in RS2 were designated rstR, rstA2 and rstB2. RS1 contains an additional ORF designated rstC. Functional analyses indicate that rstA2 is required for CTXphi replication and rstB2 is required for CTXphi integration. The amino terminus of RstR is similar to the amino termini of other phage-encoded repressors, and RstR represses the expression of rstA2. Although genes with related functions are clustered in the genome of CTXphi in a way similar to those for other filamentous phages, the CTXphi RS2-encoded gene products mediating replication, integration and repression appear to be novel. PMID- 9220001 TI - Contribution of individual promoters in the ddlB-ftsZ region to the transcription of the essential cell-division gene ftsZ in Escherichia coli. AB - The essential cell-division gene ftsZ is transcribed in Escherichia coli from at least six promoters found within the coding regions of the upstream ddlB, ftsQ, and ftsA genes. The contribution of each one to the final yield of ftsZ transcription has been estimated using transcriptional lacZ fusions. The most proximal promoter, ftsZ2p, contributes less than 5% of the total transcription from the region that reaches ftsZ. The ftsZ4p and ftsZ3p promoters, both located inside ftsA, produce almost 37% of the transcription. An ftsAp promoter within the ftsQ gene yields nearly 12% of total transcription from the region. A large proportion of transcription (approximately 46%) derives from promoters ftsQ2p and ftsQ1p, which are located inside the upstream ddlB gene. Thus, the ftsQAZ genes are to a large extent transcribed as a polycistronic mRNA. However, we find that the ftsZ proximal region is necessary for full expression, which is in agreement with a recent report that mRNA cleavage by RNase E at the end of the ftsA cistron has a significant role in the contol of ftsZ expression. PMID- 9220002 TI - Mutations in the terminase genes of bacteriophage lambda that bypass the necessity for FI. AB - DNA maturation in bacteriophage lambda is the process by which the concatemeric precursor DNA is cleaved at sites called cos to generate mature lambda DNA molecules. These DNA molecules are then packaged into procapsids, the empty capsid precursors. The enzyme that catalyses these events is lambda DNA terminase. It is composed of two subunits, made of 181 and 641 amino acids, the products of genes Nu1 and A, respectively. The product of the FI gene (gpFI) stimulates the formation of an intermediate in capsid assembly called complex II, which contains a procapsid, terminase and DNA. The mechanism of stimulation remains unknown. It has been suggested that gpFI may also stimulate terminase mediated cos cleavage, in the absence of procapsids, by increasing enzyme turnover. Mutants in FI fail to mature and package DNA but, in comparison with other capsid gene mutants, FI mutants are leaky. Second site mutants of FI phages, called 'fin' (for FI independence), bypass the necessity for gpFI. These mutants were originally localized to the region of Nu1 and A and are of two classes: finA includes those that induce the synthesis of fourfold more gene A product (gpA) than wild-type phages, and finB includes those that produce normal amounts of gpA. Whereas all finA mutants analysed map to Nu1, finB mutants have been found both in E and in Nu1. The existence of E mutants able to bypass the necessity for gpFI in vivo shows that gpE and gpFI interact, directly or indirectly. Here we have analysed and sequenced two finA mutants and one finB mutant. All of these map in Nu1. Of the two finA mutants, one corresponds to an Ala163Ser change and the other is a silent mutation. It is likely that the finA mutations alter mRNA conformation in a manner that results in an increase in the efficiency of A mRNA translation. The fourfold increase in gpA synthesis translates into a 10-fold increase in terminase activity. These results show that terminase overproduction is sufficient to bypass the necessity for gpFI and that such an overproduction can be achieved by changes in the efficiency of translation of A due to subtle changes in the sequence upstream of the gene. The finBcs103 mutation is a His-87-->Tyr change in Nu1. Therefore, an alternative way in which to bypass the requirement for gpFI involves an alteration in the structure of gpNu1. It is likely that the altered gpNu1 would increase cleavage and packaging efficiency directly or indirectly. We have determined that DNA cleavage in vivo does not occur in the absence of gpFI. Therefore it seems that gpFI somehow facilitates an otherwise latent capacity of terminase to autoactivate its nucleolytic activity. PMID- 9220004 TI - DNase I footprinting, DNA bending and in vitro transcription analyses of ClcR and CatR interactions with the clcABD promoter: evidence of a conserved transcriptional activation mechanism. AB - In Pseudomonas putida, benzoate and 3-chlorobenzoate are converted to catechol and 3-chlorocatechol, respectively, which are then catabolized to tricarboxylic acid cycle intermediates via the catBCA and clcABD pathways. The catBCA and clcABD operons are regulated by homologous transcriptional activators CatR and ClcR. Previous studies have demonstrated that in addition to sequence similarities, CatR and ClcR share functional similarities which allow catR to complement clcR. In this study, we demonstrate that CatR activates the clcABD promoter in vitro without inducer, but more transcript is produced when inducer is added. DNase I footprinting and DNA-bending analyses demonstrate that CatR binds to and bends the clcABD promoter to the same angle as does ClcR plus its inducer, 2-chloromuconate. This implies that CatR binds to the clc promoter in its active conformation. Transcription of the clcABD promoter by the alpha subunit truncation mutant (alpha-235) of RNA polymerase was sharply reduced, indicating that the alpha-subunit C-terminal domain is important. However, a small amount of transcript was produced under these conditions, indicating that other contact sites on the RNA polymerase may play a role in activation. PMID- 9220003 TI - Identification and molecular cloning of a gene encoding a fibronectin-binding protein (CadF) from Campylobacter jejuni. AB - Campylobacter jejuni, a Gram-negative bacterium, is a common cause of gastrointestinal disease. By analogy with other enteric pathogens such as Salmonella and Shigella, the ability of C. jejuni to bind to host cells is thought to be essential in the pathogenesis of enteritis. Scanning electron microscopy of infected INT407 cells suggested that C. jejuni bound to a component of the extracellular matrix. Binding assays using immobilized extracellular matrix proteins and soluble fibronectin showed specific and saturable binding of fibronectin to C. jejuni. Ligand immunoblot assays using 125I-labelled fibronectin revealed specific binding to an outer membrane protein with an apparent molecular mass of 37 kDa. A rabbit antiserum, raised against the gel purified protein, reacted with a 37 kDa protein in all C. jejuni isolates (n = 15) as tested by immunoblot analysis. Antibodies present in convalescent serum from C. jejuni-infected individuals also recognized a 37 kDa protein. The gene encoding the immunoreactive 37kDa protein was cloned and sequenced. Sequencing of overlapping DNA fragments revealed an open reading frame (ORF) that encodes a protein of 326 amino acids with a calculated molecular mass of 36872Da. The deduced amino acid sequence of the ORF exhibited 52% similarity and 28% identity to the root adhesin protein from Pseudomonas fluorescens. Isogenic C. jejuni mutants which lack the 37 kDa outer membrane protein, which we have termed CadF, displayed significantly reduced binding to fibronectin. Biotinylated fibronectin bound to a protein with an apparent molecular mass of 37 kDa in the outer membrane protein extracts from wild-type C. jejuni as judged by ligand-binding blots. These results indicate that the binding of C. jejuni to fibronectin is mediated by the 37 kDa outer membrane protein which is conserved among C. jejuni isolates. PMID- 9220005 TI - Paracoccus denitrificans CcmG is a periplasmic protein-disulphide oxidoreductase required for c- and aa3-type cytochrome biogenesis; evidence for a reductase role in vivo. AB - Cloning and sequencing of the Paracoccus denitrificans ccmG gene indicates that it codes for a periplasmic protein-disulphide oxidoreductase; the presence of the sequence Cys-Pro-Pro-Cys at the CcmG active site suggests that it may act in vivo to reduce disulphide bonds rather than to form them. A CcmG-PhoA fusion confirmed the periplasmic location. Disruption of the ccmG gene resulted in not only the expected phenotype of pleiotropic deficiency in c-type cytochromes, but also loss of spectroscopically detectable cytochrome aa3, cytochrome c oxidase and ascorbate/TMPD oxidase activities; there was also an enhanced sensitivity to growth inhibition by some component of rich media and by oxidized thiol compounds. Dithiothreitol promoted the growth of the ccmG mutant on rich media and substantially restored spectroscopically detectable cytochrome aa3 and cytochrome c oxidase activity, although it did not restore c-type cytochrome biogenesis. Assembly of the disulphide-bridged proteins methanol dehydrogenase and Escherichia coli alkaline phosphatase was unaffected in the ccmG mutant. It is proposed that P. denitrificans CcmG acts in vivo to reduce protein-disulphide bonds in certain protein substrates including c-type cytochrome polypeptides and/or polypeptides involved in c-type cytochrome biogenesis. PMID- 9220006 TI - DNA-binding activity of the A-factor receptor protein and its recognition DNA sequences. AB - The A-factor receptor protein (ArpA) containing an alpha-helix-turn-alpha-helix DNA-binding consensus sequence at its N-terminal portion plays a key role in the regulation of secondary metabolism and cell differentiation in Streptomyces griseus. A binding site forming a palindrome 24bp in length was initially recovered from a pool of random-sequence oligonucleotides by rounds of a binding/immunoprecipitation/amplification procedure with histidine-tagged ArpA and anti-ArpA antibody. By means of further binding/gel retardation/amplification experiments on the basis of the recovered sequence, a 22 bp palindromic binding site with the sequence 5'-GG(T/C)CGGT(A/T)(T/C)G(T/G)-3' as one half of the palindrome was deduced as a consensus sequence recognized and bound by ArpA. ArpA did not bind to the binding site in the presence of its ligand, A-factor. In addition, exogenous addition of A-factor to the ArpA-DNA complex induced immediate release of ArpA from the DNA. All of these data are consistent with the idea, obtained from previous genetic studies, that ArpA acts as a repressor-type regulator for secondary metabolism and cellular differentiation by preventing the expression of a certain key gene(s) during the early growth phase. A-factor, produced in a growth-dependent manner, releases ArpA from the DNA, thus switching on the expression of the key gene(s), leading to the onset of secondary metabolism and aerial mycelium formation. PMID- 9220007 TI - Regulation of the Escherichia coli K5 capsule gene cluster by transcription antitermination. AB - The expression of the Escherichia coli K5 (group II) capsular polysaccharide requires the rfaH gene. By reverse transcriptase-polymerase chain reaction (RT PCR) it was possible to demonstrate that RfaH increases the transcription of region 2 genes by readthrough transcription from the region 3 promoter. A mutation in the rfaH gene reduced this readthrough transcription from the region 3 promoter by 10-fold as measured by quantitative RT-PCR. The region 3 promoter was mapped to 741 bp 5' of the initiation codon of the kpsM gene. Deletion and insertion mutagenesis of the JUMPstart sequence, which is 28 bp 5' of kpsM and is conserved upstream of RfaH-regulated operons and other polysaccharide biosynthesis genes, confirmed that this sequence was required for expression of the K5 antigen and for the antitermination activity of RfaH. The JUMPstart sequence could cause RfaH-dependent antitermination at other Rho-dependent terminators, suggesting that the JUMPstart sequence may function in a manner analogous to a lambda nut site. On the basis of these results we propose a model by which RfaH regulates expression of E. coli group II capsule gene clusters by allowing readthrough transcription to proceed from region 3 into region 2 and that sequences within the JUMPstart sequence are essential for this process. PMID- 9220008 TI - Evidence for autolysin-mediated primary attachment of Staphylococcus epidermidis to a polystyrene surface. AB - Biofilm formation on a polymer surface which involves initial attachment and accumulation in multilayered cell clusters (intercellular adhesion) is proposed to be the major pathogenicity factor in Staphylococcus epidermidis foreign-body associated infections. We have characterized two distinct classes of biofilm negative Tn917 mutants in S. epidermidis affected in initial attachment (class A) or intercellular adhesion (class B). mut1 (class A mutant) lacks five surface associated proteins with molecular masses of 120, 60, 52, 45 and 38 kDa and could be complemented by transformation with a 16.4 kb wild-type DNA fragment. The complemented mutant was able to attach to a polystyrene surface, to form a biofilm, and produced all of the proteins missing from mut1. Subcloning experiments revealed that the 60 kDa protein is sufficient for initial attachment. Immunofluorescence microscopy using an antiserum raised against the 60 kDa protein showed that this protein is located at the cell surface. DNA sequence analysis of the complementing region revealed a single open reading frame which consists of 4005 nucleotides and encodes a deduced protein of 1335 amino acids with a predicted molecular mass of 148kDa. The amino acid sequence exhibits a high similarity (61% identical amino acids) to the atl gene product of Staphylococcus aureus, which represents the major autolysin; therefore the open reading frame was designated atlE. By analogy with the S. aureus autolysin, AtlE is composed of two bacteriolytically active domains, a 60 kDa amidase and a 52 kDa glucosaminidase domain, generated by proteolytic processing. The 120 kDa protein missing from mut1 presumably represents the unprocessed amidase and glucosaminidase domain after proteolytic cleavage of the signal- and propeptide. The 45 and 38kDa proteins are probably the degradation products of the 60 and 52 kDa proteins, respectively. Additionally, AtlE was found to exhibit vitronectin binding activity, indicating that AtlE plays a role in binding of the cells not only to a naked polystyrene surface during early stages of adherence, but also to plasma protein-coated polymer surfaces during later stages of adherence. Our findings provide evidence for a new function of an autolysin (AtlE) in mediating the attachment of bacterial cells to a polymer surface, representing the prerequisite for biofilm formation. PMID- 9220009 TI - Studies of the repressor (BlaI) of beta-lactamase synthesis in Staphylococcus aureus. AB - Purified BlaI, the putative repressor of the beta-lactamase operon in Staphylococcus aureus, binds specifically to two regions of dyad symmetry (operators) located in the blaZ-blaR1 intergenic region. BlaI binds with similar affinity to the two regions and to the related sequence upstream of the mec gene found in methicillin-resistant strains of S. aureus, providing physical evidence for the cross-talk previously observed between these systems. A change from a lysine in the N-terminus of BlaI to an alanine or deletion of the C-terminal 23 amino acids severely reduces its DNA-binding ability, demonstrating the functional importance of both the N- and C-termini. An operator DNA-protein complex observed with crude cell lysates from repressed cells, indistinguishable from that observed with purified BlaI, was eliminated by induction of the beta lactamase operon. Furthermore, BlaI is proteolytically cleaved in response to the addition of inducer in a blaR1-dependent manner, providing primary evidence for the molecular basis of induction. Thus, BlaI is shown to be the repressor of the beta-lactamase system. PMID- 9220010 TI - Involvement of the amino-terminal phosphorylation module of UhpA in activation of uhpT transcription in Escherichia coli. AB - The UhpA protein is required for expression of the sugar phosphate transporter UhpT in Escherichia coli and is regulated by phosphate transfer from the transmembrane UhpBC sensor kinase complex. UhpA action requires the sensor kinase complex and the site of phosphorylation, Asp-54, under normal conditions, but not when UhpA is overexpressed. Directed mutagenesis of the uhpA gene allowed examination of the role of several residues of UhpA in response to phosphorylation and in transcription activation. Residues Asp-9, Asp-54, and Lys 101 are highly conserved and required for function in other response regulators. Changes at any of these residues in UhpA resulted in complete loss of phosphorylation-dependent activity, but did not affect the high-level, constitutive, UhpBC-independent expression when the UhpA variants were overexpressed. Thus, these residues are important for the response to the phosphorylation pathway but not for transcription activation. Eight independent uhpA mutants selected for activity in the absence of UhpBC function carried the F17-->V or H170-->Y substitutions. Other substitutions for Phe-17 conferred various phenotypes, ranging from inducible to high-level constitutive behaviour. Residues in helix-1 flanking Phe-17 were converted to Ala or other residues. Alanine substitutions at Val-13, Arg-14, and Leu-20 resulted in complete loss of phosphorylation-dependent activation. Change of Gly-16 to Ala had no effect, but changes to other residues resulted in loss of function. Alanine substitutions at Phe-17 and at Gln-19 resulted in high-level constitutive expression, and changes at Ala-18 and Leu-21 had only modest effects. Most interesting was the L20-->A substitution, which conferred low uhpT expression when overexpressed and interfered with action of the wild-type chromosomal allele. The combination of the L20-->A change with changes at Phe-17, Asp-54 and His-170 indicated that the trans-dominant action of L20-->A occurred at several steps. The observations that UhpA can activate uhpT transcription in its unphosphorylated state are consistent with its occupancy of low-affinity binding sites necessary for promoter function. We propose that the effect of phosphorylation of UhpA is to enhance its oligomerization on the DNA surface to extend to the low-affinity sites, and that helix-1 participates in the process of oligomer formation. PMID- 9220011 TI - 'Locked-on' and 'locked-off' signal transduction mutations in the periplasmic domain of the Escherichia coli NarQ and NarX sensors affect nitrate- and nitrite dependent regulation by NarL and NarP. AB - The Escherichia coli NarX, NarQ, NarL and NarP proteins comprise a two-component regulatory system that controls the expression of many anaerobic electron transport and fermentation-related genes in response to nitrate and nitrite. Either of the two sensor-transmitter proteins, NarX and NarQ, can activate the response-regulator proteins, NarL and NarP, which in turn are able to bind at their respective DNA regulatory sites to modulate gene expression. NarX contains a conserved 17 amino acid sequence, designated the 'P-box' element, that is essential for nitrate sensing. In this study we characterize narQ mutants that also confer altered nitrate control of NarL-dependent nitrate reductase (narGHJI) and fumarate reductase (frdABCD) gene expression. While some narQ mutations cause the constitutive activation or repression of reporter-gene expression even when the cells are grown in the absence of the nitrate signal (i.e. a 'locked-on' phenotype), other mutations abolish nitrate-dependent control (i.e. a 'locked off' phenotype). Interestingly the narQ (A42-->T) and narQ (R50-->Q) mutations along with the analogous narX18 (A46-->T) and narX902 (R54-->E) mutations also confer a 'locked-on' or a 'locked-off' phenotype in response to nitrite, the second environmental signal detected by NarQ and NarX. Furthermore, these narQ and narX mutations also affect NarP-dependent gene regulation of nitrite reductase (nrfABCDEFG) and aeg-46.5 gene expression in response to nitrite. We therefore propose that the NarQ sensor-transmitter protein also detects nitrate and nitrite in the periplasmic space via its periplasmic domain. A signal transduction model, which we previously proposed for NarX, is now extended to NarQ, in which a nitrate- or nitrite-detection event in the periplasmic region of the cell is followed by a signal transduction event through the inner membrane to the cytoplasmic domain of NarQ and NarX proteins to modulate their protein kinase/phosphatase activities. PMID- 9220012 TI - Molecular localization of the Escherichia coli cytotoxic necrotizing factor CNF1 cell-binding and catalytic domains. AB - Cytotoxic necrotizing factor type 1 (CNF1) induces, in epithelial cells, the development of stress fibres via the GTPase Rho pathway. We showed that CNF1 is able to modify Rho both in vitro and in vivo. Recombinant N-terminal 33kDa (CNF1Nter) and C-terminal 14.8-31.5 kDa (CNF1Cter) regions of the CNF1 protein allowed us to demonstrate that the N-terminal region contains the cell-binding domain of the toxin and that the C-terminal region is responsible for its catalytic activity. CNF1Nter lowered the activity of CNF1 when provided to cells before the toxin whereas CNF1Cter had no effect on CNF1 cell toxicity. CNF1Cter was sufficient to induce a typical CNF1 phenotype when microinjected into African green monkey kidney cells (Vero cells), and was able to modify Rho as previously reported for CNF1. The C-terminal domain lost its catalytic activity when deleted of various subdomains, suggesting a scattered distribution of catalytic-site amino acids. Elucidation of the CNF1 functional organization and analysis of amino acid homologies between CNFs (CNF1, CNF2), Pasteurella multocida toxin (PMT) and dermonecrotic toxin of Bordetella pertussis (DNT) allowed us to postulate that CNFs and DNT act on Rho via the same enzymatic activity located in their C-terminus, and that CNFs and PMT probably bind to analogous cell receptors. PMID- 9220013 TI - Relating primary structure to function in the Escherichia coli XerD site-specific recombinase. AB - XerC and XerD are related 298-amino-acid site-specific recombinases, each of which is responsible for the exchange of one pair of strands in Xer recombination. Both recombinases encode functions necessary for sequence-specific DNA-binding, co-operative XerC/D interactions, synapsis and catalysis. These functions were related to the primary amino acid sequence by constructing and analysing internal and C-terminal XerD deletions. An XerD derivative containing residues 1-233 was proficient in specific DNA binding, but did not interact co operatively with XerC. Deletion of a further five C-terminal amino acids abolished binding to DNA. Proteins deleted for residues 32-88 and for residues 145-159 were deficient in DNA binding. Deletion of residues 244-281, a region containing amino acids necessary for catalysis, gave a protein that bound to DNA. An XerD derivative containing residues 1-268 retained co-operative interactions with XerC; nevertheless, it did not support XerC strand exchange and was defective in XerD catalysis. Residues 1-283 retain a functional catalytic active site, though a protein lacking the five C-terminal amino acids was still unable to mediate normal in vivo recombination, indicating that these residues are needed for a function that is not directly related to DNA binding or catalysis. PMID- 9220014 TI - The Neisseria type 2 IgA1 protease cleaves LAMP1 and promotes survival of bacteria within epithelial cells. AB - Infection of human epithelial cells by Neisseria meningitidis (MC) and Neisseria gonorrhoeae (GC) increases the rate of degradation of LAMP1, a major integral membrane glycoprotein of late endosomes and lysosomes. Several lines of evidence indicate that the neisserial IgA1 protease is directly responsible for this LAMP1 degradation. LAMP1 contains an IgA1-like hinge region with potential cleavage sites for the neisserial type 1 and type 2 IgA1 proteases. Neisserial type 2 IgA1 protease cleaves purified LAMP1 in vitro. Unlike its wild-type isogenic parent, an iga mutant of N. gonorrhoeae cannot affect LAMP1 turnover and its growth in epithelial cells is dramatically reduced. Thus, IgA1 protease cleavage of LAMP1 promotes intracellular survival of pathogenic Neisseria spp. PMID- 9220015 TI - A new cytolethal distending toxin (CDT) from Escherichia coli producing CNF2 blocks HeLa cell division in G2/M phase. AB - Escherichia coli strain 1404, isolated from a septicaemic calf, carries a transferable plasmid called pVir which codes for the cytotoxic necrotizing factor type 2 (CNF2). A 4h interaction between strain 1404 and HeLa cells induced the formation of giant mononucleated cells blocked in G2/M phase. Mating experiments between strain 1404 and a non-pathogenic recipient strain demonstrated that the factor(s) encoded by pVir mediated the cell-cycle arrest. A 3.3 kb DNA fragment isolated from a DNA bank of pVir was shown to code for the factor(s) causing the cell-cycle arrest. Nucleotide sequence analysis revealed the presence of three genes encoding proteins sharing significant amino acid homology with the cytolethal distending toxins (CDTs) previously isolated from E. coli, Campylobacter jejuni and Shigella dysenteriae. Southern hybridization experiments demonstrated that the pVir of other CNF2-producing E. coli strains contained sequences related to cdt. Although the amino acid sequences amongst CDT diverged significantly, the two other CDTs previously isolated from E. coli were also able to block the HeLa cell cycle. In conclusion, this study demonstrates the mode of action of CDT and will help us to elucidate the role of this emerging toxin family in microbial pathogenesis. PMID- 9220016 TI - Structural organization and characterization of the promoter region of the rat gonadotropin-releasing hormone receptor gene. AB - The gene encoding the rat gonadotropin-releasing hormone (GnRH) receptor was isolated, and its structural organization and promoter region were characterized. The gene was found to consist of three exons that encode the receptor protein, and spanned about 20 kb. Of two genomic clones analyzed, one contained the 5' untranslated region and the first exon, and the other contained the second and third exons. The sizes of the first, second, and third exons are 625, 217, and 1476 nt, respectively. The first intron is at least 12 kb in length and is located between nucleotides 522 and 523 of the cDNA reading frame, in the middle of the fourth transmembrane domain. The second intron is about 2.5 kb and is also located in the reading frame between nucleotides 739 and 740, separating the fifth and sixth transmembrane domains. Genomic blots in combination with cloning and sequencing suggested that a single GnRH receptor gene is present in the rat genome. Primer extension indicated that the transcription start site is located 103 nt upstream of the translational start codon. A putative TATA box is positioned 23 nt in front of the transcription initiation site. The 1.8 kb 5' flanking sequence contains an SF-1 site, an AP-1 site, CCAAT sequences, a Pit-1 binding site, and a potential CRE-like sequence. To evaluate promoter activity, the 1.8 kb and two 5' deleted fragments of 1.2 and 0.6 kb were fused to the luciferase reporter gene and transiently expressed in immortalized pituitary gonadotrophs (alphaT3-1 cells) and hypothalamic neurons (GT1-7 cells), and in nonpituitary (COS-7) cells. Luciferase gene expression was significantly increased by all three fragments in pituitary and hypothalamic cells, but not in COS-7 cells. The promoter activity of the 1.2 kb fragment was higher than that of the other fragments. Forskolin and cAMP analogs increased luciferase gene expression in both alphaT3-1 and GT1-7 cells, but activation of protein kinase C by phorbol myristate acetate had no effect. These studies indicate that positive and negative regulatory elements are present within the 1.8 kb 5' flanking sequence of the GnRH receptor. Knowledge of the genomic organization and analysis of the promoter region of the rat GnRH receptor gene will facilitate the elucidation of its transcriptional control in pituitary gonadotrophs and hypothalamic neurons. PMID- 9220017 TI - Expression and characterization of recombinant rat parathyroid hormone-related peptide (1-141) and an amino-terminally-truncated analogue (38-141). AB - We have synthesized and purified recombinant parathyroid hormone related peptide (PTHrP (1-141)) and PTHrP (38-141) using an E. coli system that requires minimal purification. The cDNAs encoding PTHrP (1-141) and PTHrP (35-141) respectively were inserted into the multiple cloning site of the pTrcHis-B bacterial expression plasmid. The PTHrP encoded sequences were thereby fused at their NH2 termini to six histidine residues within the fusion protein. The recombinant plasmids were transfected into E. coli cells and PTHrP synthesis was induced by addition of 1 mM isopropyl-beta-D-thiogalactopyranoside (IPTG) at 37 degrees C. The recombinant fusion proteins were purified by binding of the histidine residues to a nickel column followed by gradient elusion and dialysis. PTHrP (1 141) was released from its fusion protein by cyanogen bromide cleavage, whereas PTHrP (38-141) was released by enzymatic digestion with enterokinase. This rapid isolation method resulted in pure PTHrP (1-141) and (38-141) as judged by SDS polyacrylamide gel electrophoresis and NH2-terminal sequence analysis. PTHrP (1 141) stimulated cAMP accumulation and mobilized intracellular calcium ([Ca2+]i) in UMR106 osteoblast-like cells, and stimulated phosphate transport in OK/E renal cells, whereas PTHrP (38-141) was inert in these bioassays. Availability of PTHrP and its NH2-terminally truncated analogue, which lacks the sequence necessary for its hypercalcemic actions, will enable their biological activities to be examined in greater detail. PMID- 9220018 TI - Novel independent and synergistic regulation of gonadotropin-alpha subunit gene by luteinizing hormone/human choriogonadotropin and gonadotropin releasing hormone in the alphaT3-1 gonadotrope cells. AB - The alphaT3-1 cells are immortalized anterior pituitary gonadotropes which express gonadotropin-alpha subunit gene. These cells contain receptors for gonadotropin releasing hormone (GnRH) as well as for luteinizing hormone (LH) which can also bind human choriogonadotropin (hCG). Like GnRH, LH and hCG can upregulate the expression of gonadotropin-alpha subunit gene. While 0.1-1.0 ng/ml hCG can upregulate, higher concentrations have no effect. However, these higher hCG concentrations can act in a synergistic manner with GnRH to increase the steady state mRNA and protein levels of gonadotropin-alpha subunit. The synergism between hCG and GnRH was mimicked by LH but not by other hormones in the glycoprotein hormone family or alpha or beta subunits of hCG, suggesting that the synergism is hormone specific and requires the conformation of native hormone. The hCG induced increase in gonadotropin-alpha subunit mRNA levels was due to a significant increase in the half-life of gonadotropin-alpha subunit transcripts from 7.8 +/- 1.0 h in the controls to 16.5 +/- 3.8 h after treatment with hCG. The GnRH induced increase in gonadotropin-alpha subunit mRNA levels was due to both a significant increase in the half-life to 26.2 +/- 3.0 h as well as a significant increase in the transcription rate of the gene (159.0 +/- 7.7% of the control). A greater increase in gonadotropin-alpha subunit mRNA levels following a combined treatment with GnRH and hCG was due to a further increase in half-life to 37.6 +/- 3.1 h as well as a greater increase in the transcription rate of the gene (295.1 +/- 24.2% of the control) as compared to the treatment with GnRH alone. In summary, we conclude that LH and hCG can independently and synergistically act with GnRH to increase the expression of gonadotropin-alpha subunit gene by transcriptional as well as by post-transcriptional mechanisms in alphaT3-1 cells. These effects may be important for the increase of LH levels during the preovulatory surge. PMID- 9220019 TI - Effect of chronic growth hormone treatment on insulin signal transduction in rat tissues. AB - Growth hormone (GH) is known to produce insulin resistance, but the exact molecular mechanism remains unclear. We have chronically treated rats with GH and observed that the levels of insulin receptor in the liver or muscle were similar in both the GH-treated and non-treated rats. Insulin-stimulated receptor autophosphorylation was unaltered in the liver, but was reduced in the muscle of rats treated with GH. Insulin receptor substrate-1 (IRS-1) and phosphatidylinositol (PI) 3-kinase protein levels decreased in the liver but not muscle of GH-treated rats. There was no change in hepatic and muscle IRS-2 concentrations. A common finding in liver and muscle was the decrease in IRS-1 and IRS-2 tyrosine phosphorylation associated with a reduction in the interaction between these substrates and PI 3-kinase. These data suggest that changes in the early steps of insulin signal transduction may have a role in the insulin resistance observed in rats exposed to an excess of GH. PMID- 9220020 TI - Functional and structural analysis of R607Q and R608K androgen receptor substitutions associated with male breast cancer. AB - We previously described an androgen receptor (AR) point mutation located in the DNA-binding domain (DBD), adjacent to another AR substitution. Both were observed in two unrelated families with male breast cancer (MBC) and partial androgen insensitivity syndrome. This work was designed to determine the potential role of these two residues by in vitro study of the consequences of these two substitutions on biological functions and their structural impact at the atomic level. Mutant ARs revealed normal androgen-binding affinities and weaker DNA binding to an isolated androgen-responsive element. In cotransfection assays the mutant ARs displayed a reduced transactivation efficiency at 0.3 x 10(-10) M. Neither binding to an estrogen-responsive element nor transactivation efficiency of an ERE reporter gene was observed. Molecular modeling revealed that Arg607 and Arg608 were partially surface-exposed and located in adjacent areas in the AR-DBD complex with DNA. This is in favor of a protein-protein interaction. It is conceivable that such an interaction could be affected by mutation of one of these two arginines. PMID- 9220021 TI - Organ-specific expression pattern of the human growth hormone/placental lactogen gene-cluster in the testis. AB - In addition to testosterone, the essential paracrine factor for spermatogenesis, a number of potential auto/paracrine regulatory substances such as beta endorphins, enkephalins, chorionic gonadotropin beta, growth hormone-releasing hormone (GHRH) and insulin-like growth factor I (IGF-I) have been identified in the testis of various mammalian species. The latter findings prompted us to investigate a possible eutopic production of GH, placental lactogen (PL) and PRL in human testes. Specific expression of testicular GH/PL mRNA (n = 20) was shown by reverse transcription-polymerase chain reaction (RT-PCR) using a pair of primers designed to non-selectively amplify any transcript of the five GH/PL genes (GH-N, GH-V, PL-A, PL-B, PL-L). In contrast to the classical sites of production, the pituitary (exclusively GH-N transcripts) and the placenta (PL-A/B > 99%, GH-V < 1%), radioactive semiquantitative restriction enzyme analysis of the PCR-products revealed, that the testis has its own organ-specific pattern of GH/PL gene expression: PL-A/B > GH-V > or = PL-L = GH-N. All three organs express the single PRL gene, and testis and placenta show the alternative splice variant GH-V2. Immunological analyses by immunofluorometric assays for hPL-A/B, hGH-N and hPRL, demonstrated significant amounts of protein hormones in all testicular cytosolic homogenates (means: hPL 1.0 ng/g, hGH 5.1 ng/g and hPRL 58.7 ng/g tissue wet weight). Most noteworthy, hPL serum levels in an elderly age-matched healthy subjects (n = 18) were < 0.02 ng/ml. The concept of purely endocrine functions of placental and pituitary-derived GH/PL needs to be reassessed, since human testicular synthesis of these molecules suggest auto/paracrine functions in the male reproductive tract. PMID- 9220022 TI - Effects of triiodothyronine and retinoic acid on glucokinase gene expression in neonatal rat hepatocytes. AB - Glucokinase (EC 2.7.1.2) first appears in rat liver two weeks after birth and increases rapidly after weaning on to a high-carbohydrate diet. We investigated the role of triiodothyronine and retinoic acid in the absence of insulin on the first expression of the glucokinase gene in primary cultures of hepatocytes from 10 day-old rats. These two hormones were able to induce a rapid accumulation of liver glucokinase mRNA, secondarily to a stimulation of gene transcription during the first 24 h of culture. Moreover, the effects of individual hormones were not additive. Finally, glucokinase mRNA stability was not modified by these hormones. This suggests that triiodothyronine and retinoic acid act on glucokinase gene at the transcriptional. PMID- 9220023 TI - Specificity of transcription enhancement via the STAT responsive element in the serine protease inhibitor 2.1 promoter. AB - The growth hormone regulated serine protease inhibitor (SPI) 2.1 and 2.2 gene promoters have been shown to contain a response element similar to the gamma interferon activated sequence (GAS) family of signal transducer and activator of transcription (STAT) response elements. We have investigated the STAT and cytokine specificity of the SPI 2.1 STAT responsive element using a luciferase (LUC) reporter construct and a cDNA complementation strategy in the COS 7 cell line. Growth hormone was found to stimulate SPI-LUC reporter gene expression via activation of STAT 5, but not STATs 1 or 3, which indicates that the SPI 2.1 STAT responsive element is STAT 5 specific. In addition to the growth hormone receptor, the receptors for prolactin and erythropoietin enhanced gene transcription via the SPI 2.1 STAT responsive element, which indicates that this element is, on the other hand, not cytokine specific. Activation of STAT 5 was also observed after growth hormone treatment of cells transfected with cDNA expression plasmids for several different truncated growth hormone receptor mutants, although this activation was less efficient than with the wild type receptor. Point mutation of individual tyrosines in the growth hormone receptor intracellular domain to phenylalanines had no significant effect on signal transduction via STAT 5. These data, taken together with results from experiments using the phosphatase inhibitor sodium orthovanadate, suggest that STAT 5 may not have an absolute requirement for specific phosphorylated receptor tyrosine docking sites. That receptor tyrosine residues in a variety of amino acid contexts, or phosphorylated Janus kinase (JAK) 2 alone, can facilitate STAT 5 activation could explain the observed lack of cytokine specificity in STAT 5 activation. PMID- 9220024 TI - Transcription of the vasoactive intestinal peptide gene in response to glucocorticoids: differential regulation of alternative transcripts is modulated by a labile protein in rat anterior pituitary. AB - Expression of the vasoactive intestinal peptide (VIP) gene is controlled by glucocorticoids in a tissue- and endocrine status-specific manner. We have investigated the molecular mechanisms that determine glucocorticoid regulation of VIP gene expression in the rat pituitary. In initial experiments, using explant cultures of rat pituitary glands, we have demonstrated that treatment with the glucocorticoid agonist dexamethasone leads to a marked increase in VIP mRNA levels. This effect was found to be selective for the larger of two alternatively polyadenylated VIP transcripts, and in addition, protein synthesis inhibitors markedly enhanced the magnitude of this response indicating that a labile pituitary protein acts to attenuate the transcript-selective response to glucocorticoids. Nuclear run-on analysis of transcription demonstrated that the effects of dexamethasone in vitro are mediated largely, if not completely, at the level of transcription. In order to investigate the role of VIP promoter sequence in the glucocorticoid response, we then demonstrated that the activity of rat VIP gene promoter/reporter constructs in GH3 pituitary cells are up-regulated by dexamethasone. This up-regulation is virtually abolished following removal of promoter sequence between -162 and -89 of the start of transcription. Using an in vitro electrophoretic mobility shift assay, we have also demonstrated that this region of the promoter binds recombinant glucocorticoid receptor protein. The results of our study therefore indicate a direct mechanism of action for the modulation of VIP gene expression by glucocorticoids, and furthermore provide evidence of a mechanism that permits selective glucocorticoid regulation of alternative VIP transcripts. PMID- 9220025 TI - Lovastatin decreases prolactin and growth hormone gene expression in GH4C1 cells through a cAMP dependent mechanism. AB - The heterotrimeric G protein Gs couples several surface ligand receptors to cAMP production, as well as to both growth hormone (GH) and prolactin (PRL) gene expression in pituitary and GH cells. It has been shown that constitutively active alpha s stimulates transient expression of both PRL- and GH- chloramphenicol acetyl transferase (CAT) constructions, which indicates that both the PRL and GH promoter regions are under the influence of signal pathways mediated by alpha s. We have previously shown that the cholesterol lowering drug lovastatin decreases both the amount of G alpha s subunit in the membrane and the adenylyl cyclase activity in GH4C1 cells. Thus, we tried to verify whether that decrease in alpha s levels could affect PRL and GH secretion, as well as the expression of PRL- and GH-CAT constructions. Since the regulation of these two genes is dependent on the pituitary specific transcription factor Pit-1, the effect of lovastatin on the expression of Pit-1-CAT constructions was also studied. Our results show that lovastatin decreased the basal expression of these three cAMP-responsive genes in GH4C1 cells, being partially reversed by the addition of mevalonate to the culture medium. This effect of lovastatin on the promoter activities of the transfected constructions was also observed in PRL and GH secretion to the medium, suggesting that this drug produces similar changes in the endogenous promoters of both hormones. Moreover, the presence of lovastatin did not prevent the response to the cAMP activator forskolin, indicating that the main effect of this drug could be exerted through upstream adenylyl cyclase. In conclusion, our data indicate that lovastatin decreases the basal expression of Pit-1 and consequently of both GH and PRL genes through a mechanism probably mediated by the decrease of G alpha s levels in the cell membrane. Taken together, these results suggest that the activity of membrane heterotrimeric G proteins regulates the basal transcription of specific cellular genes in GH4C1 cells. Moreover the effects of lovastatin may be taken into account in the study of constitutively endocrine disorders associated with an increased secretion of either PRL or GH. PMID- 9220026 TI - Hypertrehalosemic hormone in a cockroach: molecular cloning and expression. AB - Hypertrehalosemic hormone (HTH) is a neuropeptide in the adipokinetic hormone/red pigment-concentrating hormone (AKH/RPCH) family that stimulates the synthesis of trehalose, the main blood sugar of many insects. The preproHTH of the cockroach Blaberus discoidalis was cloned from the corpora cardiaca (CC), the endocrine source for HTH, and the deduced sequence and organization of preproHTH were compared with other AKH/RPCH precursors. PreproHTH mRNA was determined to be approximately 0.5 kb in length as predicted by DNA sequence analysis. Northern blot analysis of the CC, ventral nerve cord, brain and fat body detected HTH-mRNA only in the CC. Levels of the HTH transcript in the CC were determined according to age, gender and mating. The HTH message was most abundant in the CC during the first several days of adult life in both sexes, then declined by 50% and were stable. HTH-mRNA levels in the CC did not respond to mating. PMID- 9220027 TI - Overexpression of glucagon-like peptide-1 receptor in an insulin-secreting cell line enhances glucose responsiveness. AB - Glucagon-like peptide-1 (GLP-1), secreted from intestine in response to food intake, enhances insulin secretion from pancreatic beta-cells. In this study, we evaluated the effects of stably transfecting the GLP-1 receptor into an insulinoma cell line, RIN 1046-38, on basal and glucose-mediated insulin secretion and on second messenger pathways involved in insulin secretion. The GLP 1 receptor transfected cells had similar insulin mRNA levels but higher insulin content compared with parental cells. In GLP-1 receptor transfected cells, glucose (0.5 mM)-mediated insulin release was increased compared with parental cells (4.52 +/- 0.79 pmol insulin/l per mg protein x h vs. 2.21 +/- 0.36 pmol insulin/l per mg protein x h; mean +/- S.E., n = 6, P = 0.015, in transfected vs. parental cells, respectively). By hemolytic plaque assay measuring single cell insulin secretion, we observed that in the GLP-1 receptor transfected cells versus parental cells the increased insulin secretion was due to the presence of more glucose-responsive cells as well as more insulin released in response to glucose per cell. Resting intracellular cAMP was higher in the GLP-1 transfected cells (35.96 +/- 3.88 vs. 18.6 +/- 2.01 nmol/l per mg protein x h; mean +/- S.E., n = 4, P = 0.039, in transfected vs. parental cells, respectively). In response to GLP-1, both GLP-1 receptor transfected cells and parental cells showed increased cAMP levels independent of glucose. Resting intracellular calcium was the same in both parental and GLP-1 receptor transfected cells. However, more cells were responsive to glucose in the GLP-1 receptor transfected cells and the calcium transients attained in the presence of glucose developed at a faster rate and reached a higher amplitude than in parental cells. We conclude that having an excess of GLP-1 receptors renders beta-cells more sensitive to glucose. PMID- 9220028 TI - Functional expression of bombesin receptor in most adult and pediatric human glioblastoma cell lines; role in mitogenesis and in stimulating the mitogen activated protein kinase pathway. AB - Functional bombesin receptors were identified in most human glioblastoma cell lines examined (approximately 85% of lines). Bombesin stimulated the release of intracellular Ca2+ in human adult (U-373MG, D-247MG, U-118MG, U-251MG, D-245MG, U 105MG, D-54MG, A-172MG, and D-270MG lines) and pediatric (SJ-S6 and SJ-G2 lines) glioblastoma cell lines. Stimulation of the glioblastoma cell line U-373MG with bombesin or gastrin-releasing peptide (GRP) induced mitogenesis, measured by [3H]thymidine incorporation into DNA, and stimulated the tyrosine phosphorylation of the mitogen-activated protein (MAP) kinases (Erk1 and Erk2). The stimulation of the MAP kinase phosphorylation in U-373MG cells was time- and peptide concentration-dependent. Both bombesin and GRP showed similar potencies in stimulation of intracellular Ca2+ release and activation of the MAP kinase pathway in U-373MG cells, whereas neuromedin B (NMB) peptide was less potent. Bombesin and GRP induced the release of cytosolic Ca2+ in a concentration dependent manner. Because bombesin and GRP were more potent than NMB peptide in increasing the cytosolic Ca2+ levels in U-373MG cells, we concluded that the BB2 subtype (also known as GRP-preferring receptor subtype) of the bombesin receptor is expressed in this cell line. The bombesin receptor antagonist ([Leu13 psi(CH2NH)Leu14]bombesin) blocked bombesin induced Ca2+ release and attenuated MAP kinase activation in U-373MG cells demonstrating that bombesin is acting through a receptor-dependent mechanism. This study indicates that functional bombesin receptors are widely expressed in human glioblastoma cell lines. PMID- 9220029 TI - Differential induction of inositol phosphate metabolism by three adipokinetic hormones. AB - Many (in)vertebrates simultaneously release several structurally and functionally related hormones; however, the relevance of this phenomenon is poorly understood. In the locust e.g. each of three adipokinetic hormones (AKHs) is capable of controlling mobilization of carbohydrate and lipid from fat body stores, but it is unclear why three AKHs coexist. We now demonstrate disparities in the signal transduction of these hormones. Massive doses of the AKHs stimulated total inositol phosphate (InsPn) production in the fat body biphasicly, but time courses were different. Inhibition of phospholipase C (PLC) resulted in attenuation of both InsPn synthesis and glycogen phosphorylase activation. The AKHs evoked differential formation of individual [3H]InsPn isomers (InsP(1-6)), the effect being most pronounced for InsP3. 40 nM of AKH-I and -III induced a substantial rise in total InsPn and [3H]InsP3 at short incubations, whereas the AKH-II effect was negligible. At a more physiological dose of 4 nM, the AKHs equally enhanced Ins(1,4,5)P3 levels. The InsP3 effect was most prolonged for AKH III. These subtle differences in InsPn metabolism, together with earlier findings on differences between the AKHs, support the hypothesis that each AKH exerts specific biological functions in the overall syndrome of energy mobilization during flight. PMID- 9220030 TI - Selective modification at the N-terminal region of human growth hormone that shows antagonistic activity. AB - A new analogue of recombinant human growth hormone (hGH), hGH des(1-6,14) was expressed in Escherichia coli, refolded and purified to homogeneity. The mutation decreased the hormone's ability to bind lactogenic and somatogenic receptors through its site 1, and almost completely abolished its ability to bind these receptors through site 2, as evidenced by both binding and gel-filtration experiments. More specifically, the binding to prolactin receptors (PRLRs) from various species or their soluble recombinant extracellular domains (ECDs) was decreased 1.5-4-fold, whereas the binding to hGH receptor (hGHR) was decreased 10 85-fold. These changes caused an almost total loss of hormone agonistic activity in several in vitro bioassays and subsequently, the hGH des(1-6,14) analogue acquired antagonistic properties. This antagonistic activity was dependent upon modification of site 1. In those cases in which the binding was reduced only slightly, e.g. binding to rabbit PRLRs, hGH des(1-6,14) acted as a strong antagonist, whereas in others in which the binding of site 1 was reduced to a higher degree, such as other PRLRs and, in particular, hGHR, the antagonistic activity was correspondingly weaker. Circular dichroism spectra of the analogue suggested that these changes do not result from a decrease in overall alpha-helix content, but rather from minor local structural modifications at the N-terminus. PMID- 9220031 TI - Oct-1, silencer sequence, and GC box regulate thyroid hormone receptor beta1 promoter. AB - Thyroid hormone, acting through thyroid hormone receptors (TRs), plays a crucial role in brain development and its insufficiency results in irreversible brain damage. TR alpha mRNA is expressed continuously from early embryonic stages, but the level of TR beta1 mRNA in brain is more abundant in adult than in fetus. To identify important factors which regulate TR beta1 expression, we compared mouse fetal and adult brain nuclear extracts by DNase I footprinting and electrophoretic gel mobility shift assays (EMSA) of the TR beta1 promoter. We carried out transient transfection studies in COS 1 cells using the TR beta1 promoter fused to Luciferase gene, and used mutated promoter vectors and various expression vectors. In DNase I footprinting using the fragment -950 to -717, fetal brain nuclear extracts protected the areas -910 to -884 and -815 to -800 more than did adult extracts. In EMSA, proteins in fetal nuclear extracts bound to a silencer sequence (-924 to -916), GC box (-901 to -887), and E box (-810 to 805), more strongly than did proteins in adult brain extracts. The bands formed on GC box were not supershifted by Sp-1, Sp-2, Sp-3, Sp-4, EGR-1, or EGR-2 antibodies. Three bands were detected on the octamer binding site probe (-913 to 906) and one protein was supershifted by Oct-1 antibody. Adult brain extracts appear to contain more Oct-1 protein than do fetal extracts. The other two bands were more intense in fetal extracts than in adult extracts, but were not supershifted by either Oct-1 or Oct-2 antibodies. Mutation of the silencer response element, mutation of the GC box, and Oct-1 over expression in COS 1 cells increased TR beta1 promoter function as assayed by Luciferase reporter. Mutation of the octamer binding site, to which only Oct-1 bound in COS 1 cells, decreased Luciferase reporter activity. Thus the TR beta1 promoter was regulated negatively by the proteins bound to the silencer sequence and the GC box, and positively by Oct-1. Silencer and GC box binding proteins are more abundant in fetal brain, and Oct-1 is more abundant in adult brain. The results may be responsible for increased amounts of TR beta1 present in late fetal and adult brain. PMID- 9220032 TI - Characterization of prolactin- and growth hormone-binding proteins in milk and their diversity among species. AB - The present study was undertaken to identify and characterize the diversity and species distribution of soluble prolactin binding-protein (PRL-BP) and growth hormone-binding protein (PRL-BP) in mammalian milk. We previously divided mammalian serum GH-BP into four main groups and identified a GH-BP with shared lactogenic/somatogenic properties in rabbit, horse, dog, pig and cat (Type III species). Here we describe PRL-BP in milk of Type III species and show it is relatively conserved within the group, having similar characteristics in terms of binding affinity for hGH (0.74-5.5 x 10(10) M(-1)), specificity towards the lactogenic hormones and molecular weight (approximately 35 kDa), except for the more heterogeneous pig milk (approximately 43 to approximately 88 kDa) Furthermore, high affinity PRL-BP was also demonstrated in sheep milk, having pure lactogenic specificity and an Mr of approximately 35 kDa. Human milk contained a high affinity PRL-BP/GH-BP, which was recognized by both hPRL and hGH and also having an Mr of approximately 35 kDa. In rabbit milk a separate GH-BP was also detected; it was clearly distinguished from the corresponding milk PRL BP on the basis of its Mr of approximately 44 kDa (vs. approximately 32 kDa for PRL-BP), its shared lactogenic/somatogenic hormonal specificity (vs. purely lactogenic for PRL-BP) and also on the basis of its relative resistance to heating at 56 degrees C for up to 3 h, while PRL-BP activity was completely destroyed within 30 min. This diversity of milk PRL-BP and GH-BP among mammalian species fits in with our earlier classification of serum GH-BP and also with the reported evolutionary rates of PRL and GH; this suggests these BPs may play important species-specific roles in the suckling newborn and/or maternal mammary gland, in keeping with the functions described for GH-BP. PMID- 9220033 TI - Phenomenology in psychiatry and psychoanalysis. PMID- 9220034 TI - Etiology of the masochistic and narcissistic personality. AB - It is the premise of this article, that at least in some instances, narcissistic and masochistic characters may develop from different role assignments in the same family (i.e., families with narcissistic dynamics). It is hypothesized that the child who later becomes narcissistic becomes assigned the role of the good child, remains merged with the mother, and becomes her ego-ideal. In contrast, the mother projects the egodystonic aspects of herself onto the child who becomes scapegoated, more willful, and defiant and eventually masochistic. The author has observed among her patients that some variables that seem to have contributed to the particular role assignment of a given child in the family are birth order, temperament, gender, resemblance to grandparents or significant objects in parents' life, and/or innate talents, gifts, and differences. It is suggested that the presence of narcissistic dynamics in the families of both masochistic and narcissistic characters may account for the similarities between the two character structures previously noted in the literature. It is obviously a limitation of this article that conclusions about family members (i.e., mothers and siblings) are based on reports of patients during long-term psychoanalytic treatment rather than direct observation. It is thus recommended that future research efforts attempt to verify these hypotheses through longitudinal family studies. PMID- 9220035 TI - Conceptualizing defense mechanisms from drive theory and object relations perspectives. AB - Analyzing common defensive features from dual theoretical perspectives, repression is postulated as the underlying mechanism, and is differentiated from denial and suppression. Acting out and intrapunitiveness are interpreted as polar options within displacement. Drive theory and object relations implications are outlined in the conceptualization of intellectualization, reaction formation, and projection, particularly as they are elaborated by basic tenets in defense theory. PMID- 9220036 TI - Song synthesis: further neuropsychological assumptions of induced song recall. PMID- 9220037 TI - Nifedipine gastrointestinal therapeutic system versus nifedipine coat-core: comparison of efficacy via 24-hour ambulatory blood pressure monitoring. AB - OBJECTIVE: To assess the comparable efficacy and adverse effect profile of two extended-release preparations of nifedipine--gastrointestinal therapeutic system (GITS) and coat-core (CC)--in patients with mild-to-moderate hypertension. DESIGN: Single institution, single-blind, prospective study. SETTING: Dwight David Eisenhower Army Medical Center, Fort Gordon, GA. PATIENTS: Ninety-one patients who were taking nifedipine GITS as a sole antihypertensive agent were randomized to receive either GITS or CC. After 3 weeks, 24-hour ambulatory blood pressure monitoring was conducted and an adverse effect questionnaire was administered. The patients were then crossed over to the other treatment arm and monitoring was repeated after 3 weeks. MEASUREMENTS: Mean blood pressure, heart rates, and the percentage of readings exceeding 140 mm Hg systolic and 90 mm Hg diastolic were compared for the 24-hour period. Additionally, mean blood pressures at 4-hour intervals after drug administration and heart rate during the first 8 hours of the dosage interval were compared. RESULTS: Ninety-one patients enrolled, 79 completed the study, and 62 patients were included in the efficacy analysis. A statistically significant difference (p = 0.020) was shown only in the last 4-hour systolic blood pressure. However, this difference was small (122 +/- 15 mm Hg with GITS vs. 126 +/- 14 mm Hg with CC). There was no difference in the percentage of readings exceeding 140 mm Hg systolic or 90 mm Hg diastolic. Neither dosage nor treatment order had an effect on the results. Adverse effects were reported with a greater frequency during CC therapy (40 with CC vs. 22 with GITS; p = 0.006), but were generally transient. Discontinuation of the drug was necessary in 3 patients during the CC cycle. CONCLUSIONS: GITS and CC demonstrated clinically equivalent antihypertensive efficacy in the study population. The CC produce may have a higher rate of adverse effects, but drug discontinuation was uncommon. PMID- 9220038 TI - Inappropriate medication prescribing for the elderly by office-based physicians. AB - OBJECTIVE: To estimate the prevalence of inappropriate medications prescribed by office-based physicians for patients 65 years or older. DESIGN: A nationwide cross-sectional survey of office visits by the elderly. SETTING: The National Ambulatory Medical Care Survey (NAMCS) 1992, a national probability sample survey of office visits by ambulatory patients within the continental US. SUBJECTS: A national probability sample of patients 65 years or older visiting office-based physicians. National estimates are based on the National Center for Health Statistics weighting procedure for the NAMCS sample. MAIN OUTCOME MEASURES: Prevalence of 20 inappropriate medications that should be entirely avoided in the elderly, using criteria developed by a panel of national experts in geriatric medicine and geriatric pharmacology. RESULTS: In the US during 1992, an estimated 8.47 million (95% CI 7.66 million to 9.28 million) office visits by the elderly indicated prescribing of at least 1 of the 20 inappropriate medications. Approximately 7.75 million (95% CI 6.98 million to 8.52 million) visits by the elderly involved 1 inappropriate medication and 0.72 million (95% CI 0.51 million to 0.93 million) visits included 2 inappropriate medications. According to the NAMCS, office-based physicians prescribed at least 1 inappropriate medication to 7.58% of the elderly who received prescriptions. The most frequently prescribed inappropriate medications were propoxyphene, amitriptyline, dipyridamole, diazepam, and chlorpropamide. Elderly patients rarely received prescriptions from office-based physicians for drugs such as secobarbital, isoxsuprine, trimethobenzamide, and carisoprodol. Furthermore, office-based physicians did not prescribe cyclandelate, pentobarbital, or phenylbutazone for the elderly. CONCLUSIONS: The prescribing of inappropriate medications by office-based physicians raises concerns regarding the quality of care for the elderly in ambulatory settings. The crux of improving patient care in ambulatory settings rests with collaborative efforts between physicians and pharmacists. PMID- 9220039 TI - Methodologic assessments of quality of life measures in clinical trials. AB - OBJECTIVE: To provide a comprehensive overview and to evaluate the quality of published clinical trials assessing the effect of drug therapy on patients' quality of life. DATA SOURCE: Clinical trials that assessed the effect of drug therapy on patient quality of life published in English, peer-reviewed journals were identified through a MEDLINE search (1966-1995) and review of references from recent publications. DATA EXTRACTION: A data collection form was used to record information on trial demographics, quality-of-life assessment, study design, and statistical analyses. A quality score was computed for each article based on a checklist of items. RESULTS: Two hundred sixty-five articles were eligible for this study Reliability data on the quality-of-life instruments were provided by 23.8% of the studies and validity data were provided by 21.5%. Quality of life was defined in about 14% of the trials, while 15% provided the rationale for selecting the specific instrument(s). The average overall quality score for the trials was 0.34, based on a scale of 0-1. The trials with quality of-life scores as the primary end point had significantly higher quality scores than those designed primarily to measure clinical outcomes (p < 0.05). CONCLUSIONS: Although there was a gradual but significant improvement in the quality of published clinical trials over time, more attention should be paid to various aspects of quality-of-life assessment (e.g., defining construct, instrument selection). PMID- 9220040 TI - Pharmacoepidemiology of urinary tract infections in Iowa Medicaid patients in urban long-term-care facilities. AB - OBJECTIVE: To describe the therapeutic management of Medicaid patients with urinary tract infections (UTIs) in urban long-term-care facilities (LTCFs) and to link individual therapies to patient outcomes. DESIGN: Retrospective review of medical records in LTCFs of patients who had documented UTIs. METHODS: Patient data were collected from 17 LTCFs in the Des Moines, IA, metropolitan area during a 1-year period starting January 1, 1995. Patients with UTIs were selected from the LTCF infection control logs. Data collected on patients included demographics, concomitant diseases, type of UTI (i.e., symptomatic, asymptomatic, catheter-related), process measures for management, UTI treatment, patient outcomes, and follow-up. Patient outcome data were defined as either cure or no cure. A UTI cure was defined as a negative urine culture while taking antibiotic therapy and/or complete resolution of signs and symptoms, as well as no further treatment given within 2 weeks after the end of treatment. RESULTS: Data were collected on 310 patients who had at least one UTI over the 1-year study period. Patients were primarily elderly (mean age 82.2 +/- 12.3 y), white (95.1%), and female (83.9%). Concomitant diseases were common and about one-fourth (23.0%) of the patients were catheterized. There were 536 UTI events (the unit of analysis) documented over the 1-year period, with about one-half (45.9%) being UTIs with symptoms consistent with uncomplicated lower UTI. Nearly two-thirds (62.3%) of the patients were cured, based on the study definition; there was no association between cure and type of antimicrobial therapy (p = 0.99). Over one-third (35.2%) of the UTIs were treated with a quinolone antibiotic. Others were treated with trimethoprim/sulfamethoxazole (24.4%), nitrofurantoin (13.9%), cephalosporin (10.4%), or ampicillin/amoxicillin (9.8%). Sixty-day follow-up showed no association between type of therapy and hospital readmission, physician follow-up visits, or subsequent UTIs. CONCLUSIONS: There were no differences in cure rates when comparing LTCF UTI patients receiving various regimens. With outcomes being the same, the clinician should closely consider costs of drug therapy in selecting a treatment preference. PMID- 9220041 TI - Economic evaluation of the use of nadroparin in the treatment of deep-vein thrombosis in Switzerland. AB - OBJECTIVE: To compare the cost implications, from the payer's perspective, of the use of nadroparin instead of unfractionated heparin in the initial treatment of deep-vein thrombosis. DESIGN: Cost-minimization study. SETTING: Switzerland. MATERIAL: Survey of clinical practice in six Swiss hospitals used to model three treatment regimens. MAIN OUTCOME MEASURES: Cost of treatment ($ US) per patient. RESULTS: Treatment with nadroparin instead of unfractionated heparin would reduce costs by $153 per patient. Treatment with nadroparin instead of subcutaneous unfractionated heparin would reduce costs by $109 per patient. CONCLUSIONS: The cost of initial treatment of deep-vein thrombosis is considerably lower with nadroparin than with either of the alternative regimens. Nadroparin reduces costs through greater ease of administration and by reducing the amount of laboratory monitoring. Treatment with nadroparin might also allow patients to be discharged from the hospital more quickly than is possible with intravenous infusion of unfractionated heparin. PMID- 9220042 TI - Abuse of prescription medicines in southwestern France. AB - BACKGROUND: Few quantitative data are available concerning abuse of medicine in the general population, although dependence on prescription medicines involves a significant proportion of the population. Falsified prescription forms can be used as an indicator of abuse. METHODS: Community pharmacists in a representative network were asked to report any falsified prescription form presented over a 1 year period. Sales data were used to express results as abuse rate and abuse rate ratio. RESULTS: Two-thirds of the 130 pharmacies in the network reported at least 1 falsified prescription. The reported incidence of falsified prescriptions was 2.3 per 10 000 inhabitants. A total of 392 falsified prescription forms was collected. The abuse rate ratios were 171 (95% CI 140 to 210) for dextroamphetamine-phenobarbital in combination, 168 (95% CI 131 to 216) for fenozolone, 67 (95% CI 53 to 84) for buprenorphine, and 40.5 (95% CI 33 to 50) for clobenzorex. CONCLUSIONS: These results show the efficiency of a method for detecting falsified prescriptions forms using community pharmacists. The abuse of medicines already known for their addictive potential can be estimated and alerts can also be detected. PMID- 9220043 TI - Stability of amiodarone in an oral suspension stored under refrigeration and at room temperature. AB - OBJECTIVE: Amiodarone is currently available in a tablet dosage form, which cannot be used in young pediatric patients. The objective of our study was to determine the stability of amiodarone in an oral suspension stored at two temperatures. METHODS: Commercially available amiodarone tablets (200 mg each) were dissolved in purified water and a suspension prepared in methylcellulose 1% and syrup to yield a concentration of 5 mg/mL. The dosage form was stored in 10 glass and 10 plastic prescription bottles. One-half of the bottles were stored at 4 degrees C and the others at 25 degrees C. Three samples were taken from each bottle at 0, 7, 14, 28, 42, 56, 70, and 91 days (n = 15). Amiodarone concentrations were measured by a validated and stability-indicating HPLC method; the pH was also determined in each sample. The drug was considered stable if its concentration exceeded 90% of the original concentration. RESULTS: The mean concentration of amiodarone was 90% or more at 4 degrees C for 91 days and at 25 degrees C for 42 days. The concentration was not affected by the type of storage container. Over 91 days, the pH did not change at 4 degrees C; it decreased slightly from 4.4 to 4.3 at 25 degrees C. CONCLUSIONS: Amiodarone was stable in an oral suspension for 3 months under refrigeration and for 6 weeks at room temperature. PMID- 9220044 TI - Amlodipine overdose. AB - OBJECTIVE: To report a nonfatal intentional overdose of amlodipine. CASE SUMMARY: A 42-year-old woman with a history of hypertension reported ingesting 50-100 mg amlodipine besylate and at least 40 ounces of beer in a suicide attempt. The patient's symptoms were mild; BP ranged from 79/50 to 113/76 mm Hg and HR from 92 to 129 beats/min (sinus tachycardia). Laboratory studies revealed normoglycemia, mild metabolic acidosis, mild hypocalcemia, blood ethanol concentration of 263 mmol/L, and a serum amlodipine concentration of 88 ng/mL (normal 3-11) 2.5 hours after ingestion. Therapy included activated charcoal, whole bowel irrigation, and intravenous NaCl 0.9%. After receiving 1.5 L of NaCl 0.9%, the patient developed signs of mild pulmonary edema that resolved over several hours without intervention. A serum amlodipine concentration obtained 35 hours later was 79 mg/mL. The patient was discharged on day 2 in good condition. DISCUSSION: In this case, an amlodipine overdose was associated with sustained hypotension and sinus tachycardia, as well as transient pulmonary edema following relatively low-volume fluid replacement. A previously published report described an amlodipine overdose that was fatal due to refractory hypotension and was complicated by concomitant oxazepam overdose. CONCLUSIONS: Amlodipine overdose produces prolonged hemodynamic effects and may lead to pulmonary edema. Due to a long elimination half-life and delayed onset of effects, patients with amlodipine overdose should receive aggressive decontamination therapy and may require extended clinical monitoring and supportive care if they are hemodynamically unstable. PMID- 9220045 TI - Pharmacokinetic evaluation of a case of massive sotalol intoxication. AB - OBJECTIVE: To describe serum concentrations and clearance of sotalol after a massive overdose. CASE SUMMARY: A 37-year-old white man took 11.2 g of sotalol hydrochloride tablets in a suicide attempt. The first serum d,l-sotalol concentration 3 hours after taking the first tablet was 20.6 mg/L and the last measured concentration 59 hours later was 1.8 mg/L. Logarithmic transformation of the concentration data indicated two separate monoexponential phases in the elimination curve, with half-lives of 30.1 and 11.6 hours. DISCUSSION: The shorter serum half-life in the later phase is comparable with that in four previously reported sotalol intoxications and within the normal range. The elimination rate increased in a temporal manner with an increase in systolic blood pressure about 30 hours after the patient was admitted. Since the sotalol elimination rate depends principally on renal function, we believe the initially slow elimination is due to a temporary reduction of the renal function caused by the systolic hypotension. CONCLUSIONS: An initial phase of slow sotalol elimination may occur after severe overdoses. In our patient this was probably due to hypotension. Thus, blood pressure should be monitored carefully. PMID- 9220047 TI - Digoxin toxicity secondary to clarithromycin therapy. AB - OBJECTIVE: To report a case of digoxin toxicity thought to be secondary to clarithromycin therapy. CASE SUMMARY: A 78-year-old white woman with congestive heart failure taking digoxin 0.25 mg po qd presented to our hospital with nausea, vomiting, and diarrhea. She had taken clarithromycin 500 mg po bid for 3 days, and a serum digoxin concentration obtained the day of admission was 4.4 mug/L. An electrocardiogram (ECG) done on admission revealed ST segment changes consistent with digoxin effect and later asymptomatic, nonsustained ventricular tachycardia (NSVT). Clarithromycin was discontinued and digoxin was withheld at admission, resulting in the resolution of symptoms, ECG abnormalities, and NSVT on day 3 of hospitalization. On day 5 her serum digoxin concentration was 1.5 micrograms/L and digoxin therapy was reinstituted at a dose of 0.125 mg/d po. DISCUSSION: This is the fourth published case implicating clarithromycin as the cause of digoxin toxicity. This interaction is most likely due to clarithromycin eradication of digoxin-metabolizing gut flora, thereby increasing digoxin bioavailability. CONCLUSIONS: Approximately 10% of patients are thought to be extensive presystemic metabolizers of digoxin and may therefore be most susceptible to a drug interaction with clarithromycin. Serum digoxin concentrations in such patients should be monitored closely during clarithromycin therapy. PMID- 9220046 TI - Lovastatin-induced rhabdomyolysis possibly associated with clarithromycin and azithromycin. AB - OBJECTIVE: To describe two cases of rhabdomyolysis in patients taking lovastatin that were precipitated by the use of the newer macrolide antibiotics clarithromycin and azithromycin. CASE SUMMARIES: In each case, the patients were treated over 5 years with lovastatin and developed rhabdomyolysis that coincided with the completion of a prescribed regimen of a newer macrolide antibiotic. Following intravenous hydration and administration of bicarbonate, the patients' condition resolved without permanent' sequelae. DISCUSSION: Rhabdomyolysis is a clinical syndrome resulting from the destruction of skeletal muscle that may progress to renal failure Several drugs have been associated with rhabdomyolysis, including lovastatin, a hydroxymethylglutaryl-coenzyme A reductase inhibitor. Erythromycin is a macrolide antibiotic that may increase the risk of lovastatin induced rhabdomyolysis. To our knowledge, these cases are the first published reports of lovastatin-induced rhabdomyolysis associated with azithromycin and clarithromycin. CONCLUSIONS: The risk of drug-induced rhabdomyolysis due to the potential interaction between lovastatin and azithromycin or clarithromycin should be considered before the concomitant use of these agents. PMID- 9220048 TI - Fatal cardiac event following initiation of risperidone therapy. AB - OBJECTIVE: To describe a patient who developed fatal pulseless electrical activity following treatment with risperidone. CASE SUMMARY: A 34-year-old white woman with no history of cardiac disease was initiated on risperidone therapy for an acute exacerbation of chronic schizophrenia. The patient developed postural hypotension and the risperidone dosage was held at 2 mg bid. On day 5 of risperidone therapy, the patient developed cardiac arrest and was treated for pulseless electrical activity. Her electrocardiogram revealed a prolonged QRS interval of 160 msec and an abnormal QTc interval of 480 msec. Despite resuscitative efforts, the patient became asystolic and was pronounced dead. DISCUSSION: Adverse cardiac events are rarely associated with risperidone therapy. Prolongation of the QRS and QTc intervals have been reported to occur following two cases of presumed risperidone overdose and also in 8 of 380 patients in a double-blind study reported by the manufacturer. Although other possibilities exists, risperidone cannot be ruled out as the cause of the patient's fatal episode of pulseless electrical activity. CONCLUSIONS: Prolongation of the QTc interval with severe adverse effects remains a possibility with the use of risperidone. PMID- 9220049 TI - Delayed chronic diarrhea and weight loss possibly due to ticlopidine therapy. AB - OBJECTIVE: To report a case of delayed-onset chronic diarrhea, anorexia, and weight loss possibly complicating the use of ticlopidine. CASE SUMMARY: A 48-year old white man presented with unremitting diarrhea associated with anorexia and marked weight loss. His history was significant for transient ischemic attack, resistant hypertension, homocystinemia, and chronic renal insufficiency. The patient had tolerated ticlopidine therapy for 2 years without problems, and there had been no changes in his drug regimen for several months. No abnormality was found on detailed digestive system evaluation and all symptoms and signs promptly resolved after ticlopidine therapy was discontinued. DISCUSSION: The occurrence of diarrhea early during ticlopidine therapy should make the clinician consider the drug as the most likely cause. However, the appearance of chronic diarrhea, anorexia and weight loss years after a patient had tolerated the drug is less typical for ticlopidine. In such situations, if no other cause can be identified, cautious withdrawals of the drug should be considered. We believe that all the patient's symptoms reported here were likely due to ticlopidine. CONCLUSIONS: Clinicians should be aware that the gastrointestinal adverse effects of ticlopidine may surface years after therapy is begun. The cause of ticlopidine induced diarrhea remains unknown. PMID- 9220050 TI - Paclitaxel formulation as a cause of ethanol intoxication. AB - OBJECTIVE: To report a case of ethyl alcohol intoxication associated with paclitaxel administration. CASE SUMMARY: A patient who received a 3-hour paclitaxel infusion for metastatic breast carcinoma and developed symptoms of acute alcohol intoxication. A blood ethanol concentration drawn at the end of the paclitaxel infusion was 97.8 mg/dL (0.098%). DISCUSSION: The amount of alcohol contained in paclitaxel is discussed. A review of the literature revealed one patient series where the highest blood alcohol concentration was one-third that seen in our patient. CONCLUSIONS: Clinicians should recognize the potential for alcohol intoxication with paclitaxel administration. This is especially pertinent when higher doses are given over a short period of time. PMID- 9220051 TI - Danaparoid in the prevention of thromboembolic complications. AB - OBJECTIVE: To review the therapies used to prevent postoperative thromboembolic complications with a focus on the role of danaparoid, a new low-molecular-weight glycosaminoglycan. DATA SOURCES: A MEDLINE search was performed to identify pertinent English-language literature including studies, abstracts, and review articles. Key search terms included danaparoid, heparinoid, lomoparin, heparin, prophylaxis, thrombosis, embolism, thromboembolism, and thromboembolic and postoperative complications. The manufacturer of danaparoid was contracted for additional information related to this compound. STUDY SELECTION AND DATA EXTRACTION: All identified articles were reviewed for possible inclusion in this review. Comparisons primarily focused on data obtained from prospective, randomized, controlled, blind clinical trials. Another important consideration was the use of venography to determine the presence of deep venous thrombosis. DATA SYNTHESIS: Various therapies are available for the prevention of postoperative thromboembolic complications. Effective pharmacologic treatments currently available include adjusted-dose heparin, warfarin, aspirin, dextran, and low-molecular-weight heparins (LMWHs). Until recently, warfarin was considered the drug of choice for thromboprophylaxis in high-risk patients, including patients undergoing orthopedic surgical procedures. Because of their comparable efficacy and greater ease of use, LMWHs are gaining favor over warfarin in this patient population. In well-designed clinical trials involving patients undergoing elective total hip replacement or fractured hip surgery, danaparoid has demonstrated greater efficacy than other active treatments, including warfarin, dextran, aspirin, and heparin plus dihydroergotamine. While studies comparing danaparoid with LMWHs have not yet been published, danaparoid may be more useful in patients with heparin-associated thrombocytopenia. CONCLUSIONS: Danaparoid is an antithrombotic agent with characteristics that distinguish it from heparin and LMWHs. Based on the efficacy and safety data reviewed, danaparoid should be considered one of the drugs of choice for the prevention of thromboembolic complications in patients undergoing orthopedic hip procedures and the drug of choice for the management of any patient with heparin induced thrombocytopenia who requires anticoagulant therapy. PMID- 9220052 TI - Is there an expanded role for digoxin in patients with heart failure and sinus rhythm? A protagonist viewpoint. AB - The evidence supporting the efficacy of digoxin in patients with heart failure who are in sinus rhythm is substantial. Digoxin improves hemodynamics, exercise capacity, symptoms, and quality of life and reduces hospitalizations. All of this is accomplished with a drug that is very inexpensive and can be given once daily. Its safety has been established through the DIG trial. Although digoxin does not decrease mortality beyond that of diuretics and ACE inhibitors, it does not increase mortality, unlike many positive inotropes. Furthermore, digoxin, in addition to ACE inhibitors and a diuretic, decreases the hospitalization rate due to worsening of heart failure. From a managed care perspective, as well as that of the patient, this is of enormous benefit. A pharmacoeconomic analysis estimated that continuation of digoxin in patients with stable congestive heart failure could save the healthcare system an estimated $ 400 million, based on costs from one hospital. The issue is not whether to use digoxin in these patients, but rather, how early to initiate therapy. From some of the recent data in patients with systolic dysfunction and mild heart failure, as well as knowledge of the neurohormonal activation that occurs early in these patients, it could be suggested that early use of neurohormonal modulators, including digoxin, would decrease the progression of heart failure. Thus, rather than waiting for symptoms despite optimal doses of an ACE inhibitor and diuretic, as suggested by the AHCPR practice guideline for heart failure, initiation of digoxin therapy in patients as early as NYHA class II at a dosage that will achieve a serum concentration of 1.0 ng/mL or less should occur. With the understanding of digoxin's effect on the neurohormonal systems, its role in patients with preserved systolic function needs to be reexplored. The debate can now focus on asymptomatic patients or those with preserved systolic function. Could these patients benefit from therapy with digoxin as well? PMID- 9220053 TI - Is there an expanded role for digoxin in patients with heart failure and sinus rhythm? An antagonist viewpoint. PMID- 9220054 TI - Prevention of group B Streptococcus infection in neonates. AB - OBJECTIVE: To review the epidemiology of group B Streptococcus (GBS) infection, risk factors for infection, and clinical manifestations of disease in the neonate, as well as the role of chemoprophylaxis and immunoprophylaxis in prevention of GBS disease and current recommendations for prevention. DATA SOURCES AND STUDY SELECTION: MEDLINE searchers (1976-1997) of the English language literature. DATA SYNTHESIS: Despite clinical advances in health care in the past two decades, GBS remains a leading cause of serious neonatal infection. Most early-onset GBS infections can be prevented through the use of intrapartum antimicrobial chemoprophylaxis. Preventing GBS infection in neonates is more cost effective than treating GBS infections, and implementing prevention programs can reduce morbidity and mortality resulting from GBS disease. Many proposals have been made regarding prevention strategies; however, they have not been implemented widely and consistently in the US. To coordinate both pediatric and obstetric supported strategies, the Centers for Disease Control and Prevention (CDC) recently published recommendations for prevention of neonatal GBS disease through two possible strategies. In the first strategy, intrapartum antibiotic chemoprophylaxis should be offered to all women identified by prenatal culture as colonized and those who develop premature membrane rupture or onset of labor at less than 37 weeks gestation. The second strategy involves administration of intrapartum antibiotics to all women who develop one or more risk factors at the time of membrane rupture or onset of labor. CONCLUSIONS: GBS is difficult to eradicate, causing many women to be colonized with the organism during pregnancy and labor, thereby infecting their infant. Prevention strategies have been published for more than 10 years without successful implementation. Although optimal prevention management has not been defined, following one of two strategies recommended by the CDC can prevent the majority of GBS infections in neonates. PMID- 9220056 TI - Intranasal lidocaine for migraine and cluster headaches. AB - While lidocaine may not be effective for the relief of all cluster or migraine headaches and the pain may recur in some patients, this therapy may offer an important therapeutic alternative for certain migraine patients. Further research may provide more information, such as which headache types best respond to lidocaine, if higher concentrations of lidocaine are more effective, if lidocaine solution is more effective than lidocaine nose spray, and if other local anesthetics are as effective. PMID- 9220055 TI - The effect of mesalamine and nicotine in the treatment of inflammatory bowel disease. AB - OBJECTIVE: To characterize the usefulness of mesalamine and nicotine in the treatment of active ulcerative colitis and inactive Crohn's disease. DATA SOURCES: Citations were selected from the MEDLINE database. Only those involving human subjects, inflammatory bowel disease, and available in English were selected. STUDY SELECTION: Selection criteria of clinical trials and review articles assessing the effects of mesalamine and nicotine in active ulcerative colitis or inactive Crohn's disease and the utility of reducing steroid dependence or relapse rate. Less than 20% of the articles identified met the selection criteria. DATA SYNTHESIS: In patients with inactive Crohn's disease, mesalamine 2 g/d significantly reduced the risk of relapse in high-relapse-risk patients compared with placebo, reducing the relapse rate from 71% to 55%, but was ineffective in preventing recurrence of inactive Crohn's disease following surgical resection. Mesalamine 4 g/d was effective in decreasing weaning failure due to steroid dependence by 67%, although the relapse rate was not significant compared with placebo at the end of 12 months. Following surgical resection, mesalamine was unable to significantly reduce the incidence of recurrence compared with placebo at the end of 1 year. In patients with active ulcerative colitis, oral mesalamine 2 and 4 g/d was superior to placebo in inducing remission compared with placebo. Among patients with prior steroid of sulfasalazine treatment, rectal mesalamine 4 g hs achieved a remission rate of 78% in more than 12 weeks of therapy. Other studies have not found a dose response relationship with lower dosages of mesalamine. Whereas nicotine 15-25 mg/d administered as a transdermal patch produced greater symptomatic improvement in active ulcerative colitis compared with placebo, nicotine 15 mg/16 h produced results no different from those with placebo in maintaining remission in active colitis. Nicotine appears to have an adverse effect on the course of Crohn's disease and is not recommended. CONCLUSIONS: Mesalamine has demonstrated clinical effectiveness as a therapeutic agent in the treatment of active ulcerative colitis and inactive Crohn's disease. Although its relationship to inflammatory bowel disease has been known for many years, the usefulness of nicotine for the treatment of active ulcerative colitis requires further exploration before it can be recommended as therapeutic agent. PMID- 9220057 TI - Alternatives to estrogen for the treatment of hot flashes. AB - Postmenopausal women experiencing hot flashes in whom estrogen replacement is contraindicated have alternatives available to them; however, there is no clearly defined treatment modality. The literature addressing many of these alternatives has serious limitations, which include the small number of women enrolled and lack of comparative studies between agents. Each patient needs to be assessed in terms of her current medical status, concomitant medications, and the degree to which vasomotor instability interferes with everyday activities. The literature suggests that megestrol acetate 20 mg bid may provide significant relief. Women who opt to use megestrol acetate must be told in advance that the effects will not be felt immediately particularly if tamoxifen is used concomitantly. Clonidine and medroxyprogesterone may constitute potential alternatives, but patients may not be able to tolerate the adverse effects. Because of the lack of literature supporting their clinical use, options such as vitamin E and ginseng need to be approached cautiously. Exercise has a role in alleviating some of the complications associated with menopause, such as heart disease and osteoporosis, but its effect on neurotransmitters and hormone concentrations, and how this relates to the treatment of hot flashes have not been characterized. Patients should be told that regular physical activity, a balanced diet, avoidance of alcohol and caffeine, and stress reduction may be of additional help in decreasing vasomotor flushing. PMID- 9220058 TI - Guanfacine use in children with attention deficit hyperactivity disorder. AB - The role of guanfacine in ADHD remains unclear. It may be reasonable to initiate a trial of guanfacine in a patient who has not responded to or cannot tolerate other agents due to adverse effects or drug dependence, or in a patient who develops motor tics. However, large placebo-controlled, double-blind, comparative trials involving guanfacine, stimulants, and/or TCAs are necessary to fully determine the role of guanfacine in the treatment of ADHD. Presently, behavioral modification is considered a first-line therapy and may be sufficient in mild cases of ADHD. Pharmacologic intervention or a combination of pharmacotherapy and behavioral modification should be tried in patients who cannot be adequately controlled with nonpharmacologic treatment. The stimulants still are considered first-line pharmacotherapy; however, guanfacine may have a role as a second- or third-line agent in patients who do not respond to or cannot tolerate stimulants or TCAs. PMID- 9220060 TI - Selective prescribing in suicidal patients: a little thought can decrease the risk of future morbidity and mortality. PMID- 9220059 TI - Saccharomyces boulardii for the treatment of Clostridium difficile-associated colitis. AB - S. boulardii has been investigated in Europe and the US, and preliminary reports indicate that it is safe and effective in conjunction with vancomycin or metronidazole for the treatment of CDC, predominantly in patients who develop recurrence. S. boulardii in combination with vancomycin or metronidazole has not been shown to be more effective than either of these agents alone for treatment of a first episode of CDC. In addition, S. boulardii has not been studied in immunocompromised patients who may be at risk for developing fungemia. Ultimately, large-scale clinical studies are necessary to determine whether S. boulardii should be routinely used to treat patients with recurrent CDC. S. boulardii is currently undergoing Phase III clinical trials for CDC treatment in the US. Clinicians interested in information regarding participation in current studies may contact Biocodex Inc., in Seattle, Washington. PMID- 9220061 TI - Fluoxetine-induced serum sickness-like reaction. PMID- 9220062 TI - Rhabdomyolysis associated with haloperidol without evidence of NMS. PMID- 9220064 TI - Tolmetin-induced esophageal ulceration. PMID- 9220063 TI - Drug-food interaction with isoniazid resembling anaphylaxis. PMID- 9220065 TI - Nifedipine-induced gingival overgrowth. PMID- 9220066 TI - As nurses, are we still in our adolescence? PMID- 9220068 TI - Arthroscopic repairs of triangular fibrocartilage complex tears. AB - Technical advancements in arthroscopic wrist procedures have improved our knowledge of normal and abnormal intraarticular wrist function. Triangular fibrocartilage complex (TFCC) tears from trauma injuries are a common source of ulnar-sided wrist pain. Fortunately, the TFCC is a structure that can be evaluated and treated arthroscopically with results that are comparable to open surgical procedures. Successful arthroscopic repairs of TFCC tears depend on a coordinated team effort between perioperative nurses, orthopedic surgeons, nurse practitioners, and occupational hand therapists, as well as cooperation from patients and family members. This article reviews the anatomy and physiology of the TFCC, the biomechanics of the wrist and mechanisms of injury, and arthroscopic repairs of TFCC tears. PMID- 9220070 TI - Blood administration in perioperative settings. AB - Blood administration safety is the result of accurate identification of patients to blood products before transfusions, appropriate care and administration of blood products, and rapid recognition and intervention when adverse reactions occur. Health care institutions have an obligation to provide staff members clearly defined, comprehensive policies and procedures related to blood administration. These policies and procedures should be combined with appropriate personnel orientation and periodic training reviews, as well as oversight of blood transfusion practices within the institution. Perioperative nurses should know and follow institutional policies and procedures on blood administration exactly, being aware of the indications and functions of basic blood products, monitor patients for signs of adverse reactions to blood administration, and intervene promptly when adverse reactions are recognized. PMID- 9220069 TI - Advanced perioperative nursing elective for baccalaureate students. AB - Recruiting baccalaureate nursing students and experienced RNs into perioperative nursing is essential if RNs are to continue their presence in surgical settings. The advanced elective described in this article is the second part of a two elective sequence in perioperative nursing at La Salle University, Philadelphia. This collaborative program between nursing education and nursing practice contains both didactic and clinical components and broadens the exposure of basic baccalaureate and RN-baccalaureate nursing students to perioperative nursing. This experience increases graduates' marketability, provides perioperative staff members opportunities to recruit new nurses into their specialty, and creates a pool of potential perioperative staff nurses. PMID- 9220071 TI - Ensure your role in the future of perioperative nursing. PMID- 9220072 TI - Astute assessment by a perioperative nurse in an expanded role saves patient from malignant hyperthermia. PMID- 9220073 TI - What's the answer? What's the question? AB - Study results are only as meaningful as the research questions they answer. Good research ideas often emerge from clinical practice situations, but they need to undergo development and refinement to become researchable nursing research questions. PMID- 9220074 TI - The powdered latex glove war. PMID- 9220075 TI - Classification and epidemiology of the vasculitides. AB - The systemic vasculitides are rare inflammatory conditions of blood vessel walls. A number of different classification schemes have been published since the first in 1952. The important developments have been the recognition of dominant blood vessel size, the distinction between primary and secondary vasculitis and the incorporation of pathogenic markers such as anti-neutrophil cytoplasmic antibodies. In 1990 the American College of Rheumatology (ACR) published criteria for the diagnosis of polyarteritis nodosa, Churg-Strauss syndrome, Wegener's granulomatosis, hypersensitivity vasculitis, Schonlein-Henoch purpura, giant cell arteritis and Takayasu arteritis. Sensitivity and specificity rates varied considerably: 71.0-95.3% for sensitivity and 78.7-99.7% for specificity. The criteria were not tested against the general population or against patients with other connective tissue diseases or rheumatic conditions. Four years later the Chapel Hill Consensus Conference (CHCC) produced definitions for the major types of vasculitis, however, these have proved controversial. Comparison in unselected patients with systemic vasculitis (in particular polyarteritis nodosa and microscopic polyangiitis) has shown that the ACR criteria and CHCC definitions identify different patients. The systemic vasculitides are somewhat more common than previously believed. The overall annual incidence approaches 40/million adults. The most common form of primary systemic vasculitis is giant cell arteritis; Wegener's granulomatosis, microscopic polyangiitis and Churg-Strauss syndrome have similar incidences. Classical polyarteritis nodosa and Takayasu arteritis are very rare in the UK. PMID- 9220076 TI - Biopsy diagnosis of systemic vasculitis. AB - A definitive diagnosis of virtually all vasculitides requires histological documentation. Although each major type of systemic vasculitis may have its own unique features, variability and overlaps still exist, and histopathological specificity is rarely an absolute discriminator. The interpretation of biopsies for the diagnosis of vasculitis remains more an art than a science, and it requires full and complete correlation with historical, clinical, laboratory, and angiographic findings. PMID- 9220077 TI - Small vessel vasculitis and vasculitis confined to skin. AB - Cutaneous vasculitis is a heterogeneous group of disorders, which can be confined to the skin or may be part of an associated systemic disease. Various aetiological agents as well as conditions that mimic skin vasculitis, usually present with similar clinical features; mainly palpable purpura. The skin biopsies usually show leukocytoclastic vasculitis. This poses a great diagnositc and therapeutic challenge for the physician. The aetiologies, clinical features, diagnosis and treatment modalities for each form (drugs, infections, malignancies, systemic vasculitides, connective tissue disorders. Schonlein Henoch purpura, cryoglobulinaemia, cutaneous periarteritis nodosa, livedoid vasculitis, erythema elevatum diutinum and urticarial vasculitis) are reviewed. PMID- 9220078 TI - Systemic necrotizing vasculitis. AB - The revival of interest in systemic necrotizing vasculitis was initiated by the discovery of its association with anti-neutrophil cytoplasmic antibodies (ANCA). The close association of certain ANCA subspecificities, for example, proteinase 3 (Pr3) and myeloperxoidase ANCA, with Wegener's granulomatosis, microscopic polyangiitis and Churg-Strauss syndrome has led to their designation as 'ANCA associated vasculitides'. This article describes the common and divergent clinical and immunological features of the members of this 'new' family of systemic necrotizing vasculitis, which continues to grow with the widespread use of ANCA testing. In addition, the 'standard' treatment for systemic necrotizing vasculitis (daily 'low dose' cyclophosphamide plus glucocorticosteroids or 'Fauci's scheme') is compared with new stage and activity adapted therapeutic regimens. PMID- 9220079 TI - Large vessel vasculitis (giant cell arteritis, Takayasu arteritis). AB - Giant cell arteritis and Takayasu arteritis are separate but similar idiopathic diseases clinically characterized by constitutional symptoms, shared surrogate markers of systemic inflammation and indistinguishable granulomatous pan arteritis of large vessels. This review emphasizes and analyses changing perceptions about the diseases. Recent series suggest that aortic involvement in giant cell arteritis may be more common than was previously appreciated. The case for and against inflammatory arthritis in giant cell arteritis is discussed. Ethnic new geographical variation in Takayasu arteritis-disease expression is reviewed. New philosophies of treatment are presented for both diseases. Prognosis in giant cell arteritis and its relationship to treatment is analysed. The utility of the laboratory for diagnosis and monitoring disease activity is appraised for each. PMID- 9220080 TI - Vasculitis associated with connective tissue disease. AB - Vasculitis, one of the clinical features shared by connective tissue diseases, should be considered when signs and symptoms are observed that may result from tissue ischaemia due to damaged vessels. The lesions seem to result from specific and non-specific immunopathogenic mechanisms targeted at the vascular endothelium. Because of the therapeutic implications it is the physician's responsibility to document its presence and the extent of organ involvement. Prompt institution of immunosuppressive drugs may be lifesaving. On the other hand there are some forms of vasculitis accompanying connective tissue disease which are entirely benign. Patients with infarctions of extremities and progressive functional disturbances of the central nervous system or internal organs because of vasculitis should be treated with high dosages of corticosteroids in combination with cytostatic drugs. Remissions are frequently obtained within three to six months of initiation of treatment and can be maintained with a less aggressive treatment regimen. PMID- 9220081 TI - Other forms of vasculitis and pseudovasculitis. AB - Behcet's syndrome can involve all sizes and kinds of blood vessels. There is an association between arterial involvement and venous thrombosis. Pulmonary arterial aneurysms and neurological involvement have a definite influence on mortality. Male sex and young age are indicators of a more severe disease course. Immunosuppressive treatment early in the disease may affect the long term prognosis favourably. Patients with familial Mediterranean fever may develop manifestations of vasculitis. The most common associations are with Schonlein Henoch purpura and polyarteritis nodosa. In some patients the diagnosis of vasculitis precedes that of familial Mediterranean fever. Kawasaki disease, although rare, can be seen in adults. The coronary sequela of childhood disease can affect the prognosis later in life. Many conditions, like myxoma, cholesterol embolism, calciphylaxis may mimic vasculitic syndromes. These conditions should always be kept in mind because their pathophysiology and treatment are different from true vasculitides. PMID- 9220082 TI - Vasculitis by organ systems. AB - Systemic vasculitides, hitherto thought to be a rare clinical entity, are now rarely considered to be an uncommon disorder and patients are often seen between several departments, suffering from a non-infectious systemic disease with multi organ involvement. Systemic vasculitis not only poses a major management problem but also has a significant impact on healthcare resources. The clinical outcome of a vasculitic illness depends on a number of factors, such as aetiology of the vasculitic process, site, size and number of blood vessels affected, duration and severity of the disease and also the complications associated with the disease or its therapy. PMID- 9220083 TI - Laboratory findings in the vasculitides. AB - The primary vasculitides are diseases of unknown aetiology. They are characterized by inflammation of blood vessel walls. Measuring non-specific laboratory markers of inflammation is useful in the monitoring of patients with vasculitis. The diagnostic specificity of these markers is, however, restricted. In the last decade, autoantibodies reacting with myeloid granule proteins have been detected in the sera from patients with Wegener's granulomatosis, microscopic polyangiitis, Churg-Strauss syndrome, and the renal limited form of these vasculitides (i.e. idiopathic rapidly progressive glomerulonephritis). Anti neutrophil cytoplasmic antibodies (ANCA) in the aforementioned disorders react with proteinase 3 (Pr3) or myeloperoxidase (MPO), and only incidentally to other antigens such as elastase and bactericidal-permeability increasing protein. The presence of ANCA alone, in particular perinuclear ANCA, as detected by indirect immunofluorescence, has a low specificity for those vasculitides. However, in combination with the presence of anti-Pr3 or anti-MPO antibodies as detected by enzyme-linked immunosorbent assay, sensitivity and specificity for the vasculitides is high. Several in vitro and in vivo data have suggested a pathophysiological role for anti-Pr3 and anti-MPO in the associated disorders. Measuring levels of the autoantibodies seems useful for the follow-up of patients with these vasculitides. The sensitivity and specificity of rises in ANCA levels for ensuing relapses appears somewhat lower than previously suggested. Refinement of the assays, for example, by measuring subclasses and functional characteristics of the autoantibodies, may improve their value in monitoring patients with vasculitides. PMID- 9220084 TI - Disease assessment and management of the vasculitides. AB - The improvement in survival with chemotherapy has resulted in a change of the natural history of the systemic vasculitic syndromes. The vasculitides are now viewed as chronic disease rather than fatal conditions. Their course is frequently characterized by relapse as well as the scars of irreversible organ damage from disease and drug toxicity. Assessment tools are available which can serve as outcome measures in clinical trials as well as a guide to better management of individual patients. Improvements in therapy in future are dependent on a better understanding of the pathogenesis of these conditions and the ability to assess disease accurately. PMID- 9220085 TI - Confabulation, memory deficits, and frontal dysfunction. AB - This paper explores potential cognitive deficits underlying confabulation of patient, G.S., following an anterior communication artery aneurysm. G.S.'s performance on tasks assessing memory for temporal duration, temporal order, and speaker identification is examined as is his recollection of autobiographical events. We compare G.S. with three nonconfabulating patients matched with him for age, education, and neuropsychological measures of memory and frontal deficits and with three age- and education-matched control subjects. Like frontal control patients, G.S. underestimated temporal durations and showed poor source monitoring (speaker identification). In addition, G.S. showed an even more pronounced deficit in recall of autobiographical memories and relatively more detailed reports of laboratory-induced memories for imagined events. We suggest that this configuration of deficits rather than any single factor accounts for G.S.'s tendency to confabulate. PMID- 9220086 TI - A study on the emotional-processing of visual stimuli through event-related potentials. AB - The effect of emotional charge of visual stimuli on cerebral activity was investigated through ERPs. This emotional charge is explained through two dimensions: arousal (relaxing-activating) and valence (attractive-repulsive). Stimuli were 12 paintings selected through questionnaires: three activating attractive pictures (A+ group), three activating-repulsive (A-), three relaxing (R), and three neutral (N). The ERPs were recorded from the 31 subjects at F3, Fz, F4, C3, Cz, C4, P3, Pz and P4. N200 and P300 did not show significant reactions to the emotional charge of the stimuli. N300 showed greater amplitudes in response to activating stimuli: at frontal sites for A+ and at parietal sites for A-. PMID- 9220087 TI - Role of the striatum, cerebellum, and frontal lobes in the learning of a visuomotor sequence. AB - This study was designed to examine the role of the striatum, cerebellum, and frontal lobes in the implicit learning of a visuomotor sequence. The performance of patients with idiopathic Parkinson's disease (PD), with damage to the cerebellum, or with a circumscribed lesion to the frontal lobes was thus compared to that of separate groups of matched normal control subjects on an adapted version of the Repeated Sequence Test. This paradigm consists of a visual reaction-time task with a fixed embedded sequence of finger movements to be performed based on presentation of visual stimuli. Subjects received four blocks of trials (i.e., 40 presentations of a 10-item sequence) per day over 6 training days. Following the last experimental session, subjects were also given two tests measuring their declarative knowledge of the sequence. Only PD patients with a bilateral striatal-dysfunction or patients with lesions to the cerebellum failed to improve their performance in the last three training sessions, hence suggesting an impairment late in the acquisition process. Further analyses revealed that such impairment was mainly implicit in nature, and that it could not be ascribed to a general decline in cognitive functioning, to mood disturbances, or to the severity of the motor symptoms. By contrast, the level of declarative knowledge of the sequence did not differ between the three clinical groups and their respective groups of normal subjects. These findings suggest that, unlike declarative memory, the incremental acquisition of a new visuomotor skill depends upon the integrity of both the striatum and the cerebellum, but not of the frontal lobes. PMID- 9220088 TI - Alexithymia: a right hemisphere dysfunction specific to recognition of certain facial expressions? AB - The most prominent features of alexithymic people are a demonstrated reduction in the ability to identify and to describe their own feelings. In recent years, these characteristics have been related to a functional disturbance of the right cerebral hemisphere. This should result in a number of other observable effects. The present study investigated whether high and low alexithymics from a nonclinical population differed in the degree of leftward perceptual bias on chimeric tasks. The chimeras consisted of pictures of faces made of up conjoined emotive and nonemotive halves as well as asymmetrically distributed stars. Differences between high and low alexithymics in the recognition of facial expressions of emotion of whole faces were also examined. High scorers on a test of alexithymia showed overall less leftward perceptual bias than low scores on the chimeric tasks and poorer recognition of facial expressions of whole faces. There was little evidence that the reduced left bias was specific to processing of emotional expressions only, or that differences in processing of facial expressions were emotion specific. These results are argued to support the right hemisphere dysfunction model of alexithymia. PMID- 9220089 TI - Visuospatial ability of parkinsonians and elderly adults in location memory tasks. AB - Two experiments addressed the question of the spatial deficits of Parkinson disease (PD) patients, using a spatial location task which varied the characteristics of the task along an effortful continuum. In the more effortful task, 11 PD patients, 10 elderly control subjects, and 13 young control subjects were given 3 min to learn the layout of 12 places labeled on a map and then reproduce it. In the less effortful task, 9 new PD patients, 9 new elderly control subjects, and 10 new young control subjects were given 3 min to learn the layout of 12 black dots and then asked to reproduce it. In both cases the task was repeated twice. The results showed that PD patients were less accurate than young and elderly control subjects in the less effortful task. In contrast, the performance of PD patients and elderly control subjects were equivalent in the more effortful task. These results support the idea of a specific visuospatial deficit in Parkinson's disease. Moreover, the deficit in effortful tasks seems to be due to normal aging. PMID- 9220090 TI - Encoding-retrieval interactions in mild Alzheimer's disease: the role of access to categorical information. AB - Normal old adults and patients in an early phase of Alzheimer's disease (AD) were presented with photographs of common objects under two different encoding conditions: naming and naming along with category decisions. Memory was assessed with free recall, category cued recall, and recognition. For both groups, recognition was superior to cued recall which was higher than that for free recall. Most importantly, cue utilization in AD was optimized in the naming + category decision condition, although the normal old showed equivalent gains from cues following both encoding conditions. These results suggest that AD patients require more cognitive support at encoding than normal old adults to make effective use of retrieval cues. Dementia-related deficits in processing categorical information spontaneously may underlie the observed group differences in patterns of performance. PMID- 9220091 TI - The significance of body part as tool errors in limb apraxia. AB - When pantomiming to command, individuals with left hemisphere brain damage (LBD) often produce errors in which they use a body part as if it were the tool (BPT). Some clinicians question the significance of this type of error because subjects without brain damage at times also make BPT responses. We analyzed BPT errors in LBD and normal subjects who were reinstructed to modify the inappropriate BPT responses when they occurred. We also analyzed errors in normal subjects who were never reinstructed if a BPT occurred. Whereas LBD subjects who were reinstructed produced significantly more BPT errors than normals who were also reinstructed, LBD subjects were not different from normals who were not reinstructed. When reinstructed, normal control subjects correctly modified virtually all BPT errors, whereas LBD subjects did not modify BPT errors. These findings underscore the need for reinstruction when a BPT error occurs to determine whether it represents a true BPT error, a sign of limb apraxia. PMID- 9220092 TI - The influence of center of mass effect on the distribution of spatial attention in the vertical and horizontal dimensions. AB - Normal subjects attend toward the middle of grouped items (center of mass effect). In order to learn if mass effect could influence performance on line bisection tasks and if the spatial orientation of the line (vertical vs. horizontal) could influence center of mass effect, we administered bisection tasks to 16 normal subjects using either lines composed of two unequal segments (one thick and one thin) or unsegmented lines. When the longer segment was to the right, left, up, or down, subjects erred by deviating their bisection toward the longer segment (center of mass effect). Our results demonstrate that the center of mass effect can be seen with bisection tasks and is greater in the vertical than in the horizontal dimension. PMID- 9220093 TI - Differences between Alzheimer's disease and vascular dementia on information processing measures. AB - This study evaluated information processing differences between 30 vascular dementia (VaD) patients, 30 Alzheimer's disease (AD) patients, and 30 normal elderly (NE) controls. They were administered a complex reaction time test, a continuous performance test (CPT), and a neuropsychological battery. Compared to NE, both dementia groups had significantly slower motor reaction times and made more errors on the CPT. Compared to AD patients, the VaD patients were slower in stimulus categorization time and had increasing omission errors and persistent commission errors throughout the CPT trial, VaD, which usually includes frontal subcortical circuit injury, can impair mental speed and stimulus response initiation. PMID- 9220094 TI - Hemispheric asymmetries for spatial frequency discrimination in a selective attention task. AB - Hemispheric specialization for spatial frequency processing was investigated by measuring reaction times to sinusoidal gratings in 12 healthy subjects. Stimuli of 1.5, 3, and 6 c/deg were randomly presented at two peripheral locations in the left (LVF) and right (RVF) upper visual hemifields during a selective attention task. Subjects were instructed to pay covert attention and to respond to a frequency in a given hemifield ignoring all other stimuli. Results showed that RTs were significantly faster at LVF than RVF for low frequency gratings, and at RVF than LVF for high frequency gratings. Furthermore, RTs were faster to 6 than 1.5 c/deg at the RVF, while there was not a significant difference at the LVF. In our view, these findings in a task requiring fast and accurate spatial frequency discriminations may be interpreted in terms of a hemispheric asymmetry for spatial frequency processing. PMID- 9220095 TI - Treatment of grade III acromioclavicular separations. Operative versus nonoperative management. AB - Twenty-six patients with Grade III acromioclavicular joint separations were evaluated to determine the outcomes of nonoperative and operative management. Evaluation consisted of a detailed functional questionnaire, physical examination, and comprehensive isokinetic strength assessment. The patients were divided into two groups: operative (n = 16) and nonoperative (n = 10). Operative management consisted of coracoclavicular stabilization with heavy suture material and with nine of the sixteen patients treatment also consisted of coracoacromial ligament transfer and lateral clavicle resection. Nonoperative management consisted of short-term immobilization with early range of motion and rehabilitation. The two groups were similar in all characteristics except mean age: 30.7 years for the operative group and 49.6 years for the nonoperative group. Follow-up evaluation was performed an average of 32.9 months after either injury (nonoperative group) or surgery. Our results indicated that nonoperative management was superior to operative management with respect to time to return to work (0.8 months vs. 2.6 months), time to return to athletics (3.5 months vs. 6.4 months) and time of immobilization (2.7 weeks vs. 6.2 weeks). However, operative management was superior to nonoperative management in the following parameters: time to attain completely pain-free status, the patient's subjective impression of pain, range of motion, functional limitations, cosmesis, and long-term satisfaction. There were no significant differences between the two groups with respect to shoulder range of motion, manual muscle testing, or neurovascular findings. Isokinetic strength testing of the involved shoulder, expressed as a percentage of the uninvolved shoulder, showed no significant differences in peak torque, total work, or total power between the operative and nonoperative groups. However, comparison of the involved to the uninvolved extremity within each group did reveal a trend toward decreased peak torque, work, and power for abduction in the involved extremity regardless of the treatment used. These findings reached statistical significance only for power at the slower testing speed (60 degrees/sec). There was also a significant decrease in power in the involved extremity for external rotation at the faster speed (120 degrees/sec) in the nonoperative group. Finally, the absolute values for peak torque, work, and power were significantly greater for all motions tested in the operative group as compared to the nonoperative group. This may reflect the difference in age between the two groups. Based upon the patients studied, there are benefits to both nonoperative and operative methods of treatment of Grade III acromioclavicular separations. Recovery of strength did not differ between the two groups and therefore should be viewed as a less important factor in patient selection for operative versus nonoperative management. Careful patient selection should remain an important aspect of treatment for this controversial injury. PMID- 9220096 TI - Mennen plate in hip and shoulder joint replacement. AB - Sixteen patients complicated by femoral fracture and aseptic loosening with severe bone loss and two patients complicated by humeral fracture after hip and shoulder joint replacement, respectively, were collected from the Carmel and Ichilov teaching hospitals. All patients underwent internal fixation with the Mennen plate or in conjunction with revision joint replacement, either for internal fixation or for bone graft support. The results were satisfactory in fifteen patients with respect to functional activity, pain, and the radiographic evaluation. In three patients who underwent their fourth revision, the results were unsatisfactory but were better than their preoperative status. Our conclusion is that Mennen plate fixation provides a sufficient and easy internal fixation technique for a fracture around or below the un-dislodged stem of hip and shoulder in joint replacement. In an unstable femoral fracture or in a case of aseptic loosenings with marked bone loss, we recommend Mennen plate fixation in conjunction with long stem revision total hip replacement and bone graft. PMID- 9220097 TI - Improving the distal fixation of intramedullary nails in osteoporotic bone. AB - Twenty-four mildly osteoporotic human femurs were used to examine the fixation stability of several types of distal locking screws in osteoporotic femoral condylar segments. The fixated femurs were axially loaded to 1000 N at a rate of 100 N/sec to assess the locking screws' resistance to motion and then sinusoidally cycled to 10,000 cycles with a 500 N load to assess axial stability. Of the four screw configurations tested, the standard screws and the standard screws placed at a 30 degree crossing angle were significantly more stable (p < 0.05) than the step screw and osteoporotic bolt. PMID- 9220098 TI - Structural peculiarities of the tissue formed in modelling of the articular surface. AB - A model of articular cavity with smooth cover was produced by influence of shape forming element on regenerating bone tissue. Morphological, ultrastructural, histochemical, and biochemical peculiarities of the tissue of the formed articular surface are described. PMID- 9220099 TI - Reliability evaluation of classifying radial head fractures by the system of Mason. AB - Inter-observer and intra-observer reliability for classifying radial head fractures by the system of Mason was analyzed. Twenty-three cases of isolated radial head fractures and twenty-five sets of corresponding AP and lateral radiographs representing these fractures were assembled. The cases were reviewed and assessed independently according to the system of Mason by twenty practicing orthopedic surgeons. On two occasions, the inter-observer and intra-observer variation was analyzed by standard unweighted Kappa statistics. In both observations, complete agreement was seen in only 16% of the cases. Kappa statistic values indicated that 69% of the cases at first observation and 45% of the cases at second observation suggest moderate to poor agreement. Intra observer agreement between the first and second observation was graded fair to poor in 60% of the cases. Individual observer consistency was, on average, only 78% (range 60% to 92%). The demonstrated wide degree of variation suggests that the Mason classification is unreliable. PMID- 9220101 TI - A new technique for stabilization of complex intertrochanteric hip fractures. PMID- 9220100 TI - The Allen test. A study of inter-observer reliability. AB - The Allen test was performed by four physician examiners on 200 hands of healthy volunteers. Positive results were found in 5.5% of hands. There was not a single case in which all four observers agreed. Considerable inter-observer disagreement is associated with the Allen test. PMID- 9220102 TI - The immediate treatment of pelvic ring disruption with the pelvic stabilizer. AB - The management of the hemodynamically unstable patient with a severe pelvic ring disruption remains one of the most serious trauma emergencies. Standard resuscitation protocols may include attempted closure of the pelvic ring by the use of pneumatic anti-shock trousers, external fixation applied in the operating room, or a sheet wrapped around the patient in the emergency room. We report a case of pelvic ring disruption in which a successful clinical outcome was achieved with the emergent use of the Pelvic Stabilizer in the emergency room. The Pelvic Stabilizer is a device that can be effectively applied in the emergency room for the acute reduction and early stabilization of the displaced pelvis in a hemodynamically unstable patient. The use of a pelvic clamp can also be effective in the acute setting for a stable trauma patient with pelvic ring disruption. It rapidly reduces and stabilizes a potential cause for patient decompensation without obstructing access to further concomitant diagnostic or therapeutic interventions in the abdomen and perineum. PMID- 9220103 TI - A new technique for removal of intraarticular bullet fragments from the femoral head. AB - Removal of foreign bodies from a joint usually involves an extensive surgical approach. The necessity for intraarticular bullet removal has been well documented in the literature. The conventional approach for bullet extraction usually requires an open arthrotomy, arthroscopic removal or, in most cases, a combination of the two. This report involves a previously undocumented technique for bullet removal from the hip. A Synthes DHS Triple Reamer was inserted through a limited lateral incision over a guide pin that had been placed under fluoroscopic guidance. The tip of the guide pin was positioned in contact against the bullet fragments in both the anteroposterior and lateral planes. The fragments were then removed through the reamed canal. This technique allows for bullet removal without the inherent risks associated with an open arthrotomy and without the special skills required for hip arthroscopy. It is relatively easy to perform and may prove to be a valuable tool in the arsenal of orthopedists who deal with specific gunshot wounds to the hip region. PMID- 9220104 TI - Squamous cell carcinoma of the perionychium. AB - Squamous cell carcinoma of the perionychium is a rare, slow-growing tumor which is frequently misdiagnosed. Confusion arises because many problems affecting the soft tissue at the fingertips are clinically similar. Physicians should be aware of the possibility of carcinoma when faced with a chronic inflammatory process unresponsive to treatment. Diagnosis is confirmed by biopsy. Treatment options include amputation of the distal digital segment and Mohs microscopically controlled excision. PMID- 9220106 TI - The extensor digitorum brevis manus. A case report. AB - A case of a soft tissue mass of the dorsum of right hand in a 25-year-old, right handed male is described. The preoperative diagnosis was a ganglion. On exploration the mass was found to be an atavistic extensor digitorum brevis manus replacing the extensor indicts proprius tendon. Division of the extensor retinaculum resulted in improvement of the symptoms. This muscle has been encountered rarely and few cases have been reported in the literature. It presents as a mass on the dorsum of the wrist. The symptoms are pain after activity. It is nearly always diagnosed as a ganglion preoperatively. The authors conclude that extensor digitorum brevis manus muscle should be included in the differential diagnosis of soft tissue tumors of dorsum of hand. Symptomatic patients are treated by division of the extensor retinaculum. PMID- 9220105 TI - Bilateral, asymmetrical congenital dislocation of the radial heads in trisomy 8 syndrome. AB - Congenital radial head dislocation (CRHD) can occur as an isolated abnormality, as part of an upper-limb anomaly or as a feature of at least fourteen syndromes. The dislocation may be unilateral or bilateral, and rarely can be bilaterally asymmetrical. CRHD is often asymptomatic, and may go undiagnosed and remain undetected until after a radiography has been obtained for an incidental injury. It is therefore important to be able to differentiate congenital from traumatic dislocation of the radial head. We report a mentally retarded female, known to have trisomy 8, who presented with stiffness of her elbow joints and no history of preceding trauma. Radiographs confirmed bilateral asymmetrical radial head dislocation. This combination of anterior and posterior CRHD co-existing in the same patient has not been described previously with trisomy 8 syndrome. PMID- 9220108 TI - Affective disorders: discovery and recovery. PMID- 9220107 TI - Ipsilateral fractures of the proximal ulna and distal radius (Colles type). AB - A 43-year-old female fell backward outside her home while carrying a watering can. She sustained an ipsilateral fracture of the proximal ulna and distal radius (Colles fracture). The ulnar fracture was plated, and the Colles fracture was managed by closed reduction and percutaneous Kirschner wiring. The patient recovered without incident. This association of forearm fractures is unusual. Both fractures can be caused by a fall on the hand with the wrist in mid extension and the elbow flexed, and they probably occurred in quick succession. PMID- 9220109 TI - A review of functional neuroimaging in mood disorders: positron emission tomography and depression. AB - OBJECTIVE: To examine the progress of positron emission tomography (PET) as a tool for understanding the psychobiology of mood disorders, particularly major depression and bipolar disorder. METHOD: Review of the literature on functional imaging of mood disorders. RESULTS: Functional imaging techniques have been used in psychiatric research as a noninvasive method to study the behaviour and function of the brain. Techniques used so far have involved the manipulation of emotion in healthy volunteers, the evaluation of depressed (unipolar and bipolar as well as secondary depression), manic, and normal subjects under resting and various activation conditions, such as cognitive activation, acute pharmacological challenge, and chronic thymoleptic treatments. As a result, functional imaging studies tend to support abnormalities in specific frontal and limbic regions. CONCLUSION: Different PET methods demonstrate consistent abnormalities in the prefrontal, cingulate, and amygdala regions. These findings are in agreement with past animal and clinical anatomical correlates of mood and emotions. PMID- 9220110 TI - A review of continuation and maintenance electroconvulsive therapy. AB - BACKGROUND: Many patients with major psychiatric disorders who are severely ill, medication-resistant, or medication-intolerant respond more reliably and quickly to a course of electroconvulsive therapy (ECT). The management of such patients after successful treatment with ECT is of significant importance given the high rate of relapse and recurrence of these disorders. The unmet clinical need to maintain the mental health of these seriously ill patients at an optimal level has revived the interest in ECT as an alternative prophylactic treatment. METHOD: We review the historical background of ECT and the literature that supports its use as a prophylactic treatment in various disorders and special populations. A clinical summary outlining its efficacy, acceptability, risks, cost effectiveness, and medicolegal aspects is followed by a guide for prescribing ECT for prophylactic reasons. RESULTS: Continuation and maintenance ECT (C/MECT) has been found to be efficacious, safe, well tolerated, and cost-effective. Its greatest impact has been in reducing relapse, recurrence, and rehospitalization, particularly in the management of recurrent mood disorders in the elderly. The elderly are usually refractory or intolerant to pharmacotherapy but have a good response to ECT during the index episode. Parkinson's disease (PD), schizophrenia, and obsessive-compulsive disorder (OCD), as well as affective disorders coexisting with dementia, neurological disorder, or mental retardation, have also been reported to respond to C/MECT. The outcome depends greatly on rate of compliance. Cognitive risk of C/MECT need to be further studied because the literature to date consists mostly of case reports and anecdotal evidence. Controlled studies with well-defined outcome measurements are needed. CONCLUSIONS: When planning a rational approach to the care of patients with major psychiatric disorders, clinicians should carefully consider ECT along with other alternatives. PMID- 9220111 TI - Ethical research with the mentally disordered. AB - Because of concerns about competence and voluntariness, the mentally disordered constitute a vulnerable population in the context of nontherapeutic biomedical research and, as such, are in need of protection. Despite others' concern about protecting the mentally disordered, their decision-making potential should also be respected and maximized, allowing such individuals to consent to participate in experiments subject to an evaluation of their competence to make such a decision. Competent mentally disordered persons who anticipate future incapacity should be able to issue research directives or durable powers of attorney whereby they can provide explicit consent to participate in nontherapeutic research. When he or she becomes incompetent, a substitute decision maker should be able to provide consent on behalf of the mentally disordered person within established parameters. Nontherapeutic experimentation with the mentally disordered should be permitted, but only within the boundaries of ethical permissibility delineated by legislated guidelines. At present, the legal status of substituted consent for nontherapeutic procedures is uncertain and requires legislation, which in addition to legalizing such consent, would provide guidelines for substitute decision makers and for the creation of research directives. These guidelines should include restrictions on the scope of research, obligations of researchers, rights of subjects, and responsibilities of research ethics committees (RECs). In all cases, the voluntary and informed consent of the person or substitute decision maker must be obtained. PMID- 9220112 TI - Ethical research with the developmentally disabled. AB - The developmentally disabled constitute a vulnerable population in the context of nontherapeutic experimentation. Their vulnerability is characterized by diminished decision-making capacity and by susceptibility to coercive situations that may bring voluntariness into question. The international consensus is that research involving this population should be permitted, but only if the consent of a legal guardian is obtained and appropriate safeguards are introduced. Therefore, legislation regulating the ethical conduct of research should be enacted, including provision for substituted consent in the research context. Although researchers seeking the participation of a developmentally disabled individual in a protocol must presume the person to be capable of participating in the decision, they must conduct competency assessments if the person's ability to make such a decision is in doubt. Information must be presented in such a way as to maximize the individual's contribution, and capacity must be reevaluated on an ongoing basis. In addition, research on the developmentally disabled presents specific challenges to establish competency, the selection of subjects, the characteristics of the decision maker, and the model to be used in making substituted decisions. PMID- 9220113 TI - Courts and torts: the psychiatrist preparing for trial. AB - OBJECTIVE: To outline how a psychiatric expert can do an impartial assessment and medicolegal report and then give an effective presentation in court that can sustain cross-examination. METHODS: The legal principles of litigating emotional trauma are reviewed, including proving causation, characterizing emotional suffering, assessing disability, and determining a realistic prognosis. RESULTS: Psychiatrists must understand the interplay of legal and psychiatric principles when they are asked to assess litigants who are suing for monetary compensation for a widening range of emotional injuries resulting from motor vehicle accidents, slips and falls, incest and sexual abuse of children, discrimination, unlawful dismissal, malpractice, human-made disasters, product liability, and intentional torts, to name a few. CONCLUSION: The psychiatrist can prepare his or her attitude, knowledge, and skills to give a presentation in court that will be credible, trustworthy, and dynamic. With adequate preparation, the psychiatric expert can bring an informed psychiatric perspective to the court that will have a significant impact on the outcome of the judicial deliberations. PMID- 9220114 TI - Outcome evaluation of a short-term mental health day treatment program. AB - OBJECTIVES: To determine whether a structured, 6-week mental health day treatment program was meeting its objectives and to examine the program's effectiveness with specific patient groups. METHOD: Self-report questionnaires focusing on psychiatric symptoms, assertiveness, stress management, and social functioning were completed by patients directly prior to admission (pretest), at discharge (posttest), and at 4-month follow-up. Clinician ratings, including the DSM-III-R Global Assessment of Functioning (GAF) scale, were collected. Ninety-one participants completed pre- and posttests, and 51 completed the 4-month follow up. RESULTS: The majority of the participants displayed affective disorders or adjustment disorders. There was significant reduction in psychiatric symptoms and improvement in assertiveness, social functioning, and stress management from pretest to posttest. These gains were maintained at follow-up. All diagnostic groups responded similarly, except the bipolar disorder group. CONCLUSION: These data indicate that the program was meeting its objectives and offer strong support for the usefulness of short-term day treatment for wide range of patients. The bipolar group performed differently compared with the other subsamples. The reliability of the GAF scale and when it may be most useful are discussed. PMID- 9220115 TI - The impact of Canadian Criminal Code changes on remands and assessments of fitness to stand trial and criminal responsibility in British Columbia. AB - OBJECTIVE: To evaluate the impact in British Columbia of the 1992 Criminal Code of Canada amendments dealing with remands for fitness to stand trial and not criminally responsible on account of mental disorder (NCRMD) assessments. METHOD: Information on 620 remands for evaluation of fitness to stand trial and/or NCRMD were collected from a sample obtained in British Columbia from 1992 to 1994. The data collected included length of remand order, length of evaluation, criminal charges, psychiatric diagnoses, and the decisions about fitness or NCRMD. RESULTS: Remands increased by about 20% in a 1993-1994 fiscal year compared with the previous year. The majority of evaluations continue to be conducted in an inpatient facility. The goal of a 5-day evaluation period is rarely met: only 12.5% of inpatients were released within 5 days of admission, and the average length of evaluation was about 3 weeks. The use and success of the NCRMD defence appears to be on the rise. In addition, there were some striking differences in remands from metropolitan and nonmetropolitan areas in terms of rates of referral and recommendations of unfitness or NCRMD. CONCLUSION: Results indicated that Bill C-30 has not yet had the anticipated impact on remands as inpatient evaluations continue to be the norm and evaluations typically take several weeks. Suggestions for policy reform and future research are presented. PMID- 9220117 TI - Re: Presidential address. PMID- 9220118 TI - Re: Reconcilable differences: the marriage of qualitative and quantitative methods. PMID- 9220116 TI - Choking incidents among psychiatric patients: retrospective analysis of thirty one cases from the west Bologna psychiatric wards. AB - OBJECTIVE: To determine the rate of choking incidents among the psychiatric population of 4 inpatient facilities, classifying the incidents according to their probable etiology. METHOD: All incidents recorded over 18 months were retrospectively analyzed for demographic variables, psychiatric and medical diagnoses, and drug therapy at the time of incident. Where possible, patients underwent psychiatric, neurological, and medical examination. RESULTS: Thirty-one incidents were recorded involving 18 patients at a rate of one incident every 56.32 months' hospitalization per person. One case proved fatal, one patient died several weeks after the incident from aspiration pneumonia, and 5 patients needed reanimation or the Heimlich manoeuvre. Etiological classification showed that incidents due to bradykinetic dysphagia and "fast eating" were the most numerous, even among the fatal or grave cases. CONCLUSIONS: Various simple, effective preventive measures emerge from the study. PMID- 9220119 TI - Severe extrapyramidal side effects when discontinuing clozapine and starting haloperidol. PMID- 9220120 TI - Resistant depression associated with lupus erythematosus and steroid therapy. PMID- 9220121 TI - SSRI combination treatment for depression. PMID- 9220122 TI - Treatment of factitious disorder with pimozide. PMID- 9220123 TI - A case report of factitious disorder with psychiatric complaints. PMID- 9220124 TI - Psychotropic medications and cytochrome P450 2D6: pharmacokinetic considerations in the elderly. AB - BACKGROUND: The genetically polymorphic cytochrome P450 2D6 isozyme (CYP2D6) is responsible for the metabolism of numerous psychotropic medications pertinent to the practice of geriatric psychiatry. Optimal use of psychotropics in the elderly requires a thorough understanding of the determinants of marked variability in plasma concentrations. This review article will focus on basic pharmacokinetic considerations for elderly patients when psychotropics metabolized by CYP2D6, such as nortriptyline and desipramine, are prescribed. METHOD: A MEDLINE search was conducted using the subject headings "cytochrome P450," "pharmacokinetics," and "psychotropics." Relevant articles from bibliographies were also collected. RESULTS: CYP2D6 activity does not change with age. Approximately 5% to 10% of whites are poor metabolizers for CYP2D6 and are at risk for drug toxicity. Among Asians, although the prevalence of poor metabolizers in only 1%, the distribution of CYP2D6 activity in extensive metabolizers is shifted toward lower values relative to whites. CYP2D6 activity may be impaired by inhibitors such as paroxetine and fluoxetine. Inhibition of CYP2D6 activity may result in nonlinear plasma drug concentration kinetics, as well as kinetic drug interactions when other drugs metabolized by CYP2D6 are coadministered. Among extensive metabolizers, there is considerable interindividual variation in CYP2D6 activity. Significant correlations have been reported between individual CYP2D6 activity and plasma concentrations of nortriptyline and desipramine. CONCLUSION: Clinical measurement of CYP2D6 activity may potentially assist in prediction of doses required to achieve therapeutic plasma concentrations of psychotropics metabolized by CYP2D6 in individual patients. Although CYP2D6 activity does not change with age, the pharmacokinetics of psychotropics metabolized by CYP2D6 may change because of age-associated changes in hepatic blood flow, volume of distribution, and renal elimination of metabolites. PMID- 9220125 TI - Pharmacotherapy of affective disorders in old age. AB - OBJECTIVE: To present a clinical guide for the selection of safe and effective pharmacological treatment for depressed geriatric patients. METHOD: A review of the use of antidepressants in the elderly is presented based on clinical experience and a search of MEDLINE and PsychInfo data bases. Emphasis is placed on the following newer antidepressants: fluvoxamine, sertraline, paroxetine, nefazodone, venlafaxine, and moclobemide. RESULTS: With the advent of newer antidepressants, physicians have been given a wider range of generally safer antidepressant medications. Although these medications appear to have favourable side effect profile in elderly patients, this review is limited because there are few published studies of the use of these newer antidepressants in the elderly. CONCLUSION: Depression is a common psychiatric problem in old age, but it can usually be treated successfully. Although the use of antidepressants in geriatric populations is more likely to be complicated by poorly tolerated side effects and drug interactions than in younger patients, this review should help clinicians use currently available medications to the best advantage of their geriatric patients. PMID- 9220126 TI - The treatment of psychotic disorders in late life. AB - OBJECTIVE: To review the epidemiology, phenomenology, and treatment of psychotic disorders in late life. METHOD: The literature relating to psychotic symptoms in the elderly is reviewed, with a focus on the following categories: primary psychotic disorders, mood disorders, delirium, Parkinson's disease (PD), and somatic hallucinoses (including Charles Bonnet syndrome [CBS] and musical hallucinosis). Practical clinical treatment implications are discussed. RESULTS: The prevalence of psychotic symptoms increases with age, largely because of underlying medical illnesses such as dementia, delirium, and other neurological disorders that are exacerbated by sensory deficits coupled with social isolation. Treatment with the traditional high-potency neuroleptics is complicated by extrapyramidal symptoms, and sedation, postural hypotension, and anticholinergic effects complicate the use of low-potency traditional agents. Although clozapine may have a narrow use in the treatment-resistant schizophrenia and PD, it is poorly tolerated in the elderly. Risperidone has a wider use in this population and has a favourable clinical profile (at low doses). Other new neuroleptics await more formal evaluation in the elderly. CONCLUSION: Psychotic disorders in old age have more organic associations, which cause greater difficulty in their treatment. Further Evaluation of the use of atypical agents in this elderly group is indicated. PMID- 9220127 TI - Treatment of anxiety disorders in late life. AB - OBJECTIVE: To provide a current review and synthesis of the present state of knowledge of anxiety disorders and symptoms in the elderly. METHODS: Current research derived from a MEDLINE search and references in key textbook articles and other papers were reviewed. These data were combined with the clinical empirical knowledge and experience of the authors. RESULTS: Anxiety disorders and symptoms are a common presenting problem in the elderly. Current knowledge and research findings are limited. Extrapolation from adult studies are of use, but important limitations are evident because of the nature, uniqueness, and complexity of the geriatric psychiatry patient. Comorbidity, especially with depression, medical conditions, drugs, and dementia, remains an important concept in assessment and approach to management of anxiety in the older person. Comprehensive assessment of anxiety symptoms requires consideration of physical, intellectual, environmental, and social determinants. Major anxiety disorders, as defined by DSM-IV, and anxiety symptoms are significant problems in the older adult population and are responsible for significant morbidity and cost to the health care network. CONCLUSIONS: Anxiety disorders and symptoms in old age, although common, have received little research focus to date. A comprehensive, careful approach by the clinician to assessment and management is required because anxiety is often a comorbid condition in the elderly. Effective treatments are available and should be applied in a flexible, integrated, and specific manner. PMID- 9220128 TI - Cognition-enhancing drugs in dementia: a guide to the near future. AB - OBJECTIVES: To facilitate access to the available literature and to assist clinicians with the decision to recommend the use of medications for the enhancement of cognition in dementia. METHOD: A qualitative review of published research. Methodological issues confronting research in this area are described. An organizational scheme for the medication types, based on pathophysiologic processes relevant to Alzheimer's disease (AD), is reviewed. The paper makes extensive use of tablets to present the minimal data necessary for the reader to appraise critically all of the original publications found. The paper further presents in summary form the opinions of previous reviews on each of the medications. RESULTS: We identified 45 medications in the published research in which humans with dementia were assessed as having a change in cognition. Immediate use of tacrine is supported by the evidence, but the degree of benefit is modest, and side effects are problematic. CONCLUSION: A number of medications warrant further investigation. Tacrine can be offered to patients with careful education regarding the limited efficacy and potential side effects. Newer, perhaps safer, anticholinesterase inhibitors are now becoming available. Referral to a research study examining other medications is suggested, as are some "common sense" strategies. PMID- 9220129 TI - From transmitters to treatment: the pharmacotherapy of behavioural disturbances in dementia. AB - BACKGROUND: Behavioural disturbances in dementia are a common cause of excess morbidity, impairing the quality of life for both patient and caregiver. As part of a comprehensive approach to management, which includes a search for underlying causes and behavioural interventions, pharmacotherapy can be extremely helpful in alleviating symptoms such as agitation, aggression, and psychosis. METHOD: This paper reviews recent studies that examine the neurochemical basis of these behavioural disturbances in order to provide a rationale for the various classes of psychotropics which have been used. RESULTS: While neuroleptics have been the best-studied class of drugs to date, modest efficacy and significant potential side effects often limit their use. Newer atypical neuroleptics may be better tolerated, though controlled data have yet to be published. There is increasing support of the use of carbamazepine and antidepressants such as trazodone and the selective serotonin reuptake inhibitors (SSRIs). CONCLUSION: Further controlled studies of all of these agents are required. In order to determine whether transmitter-specific or behavioural-specific targeted interventions truly provide a rationale for the effective pharmacotherapy of these disorders. PMID- 9220130 TI - Extra-label drug use--an addendum. PMID- 9220131 TI - Prevalent diseases of ostrich chicks farmed in Canada. AB - In Canada, ostriches are now slaughtered for their meat and hides. The mortality rate in ostrich farming is highest in chick units. An increased chick survival rate impacts positively on production and profit. This paper will focus on common health disorders that affect chick production costs. These are discussed under the following categories: digestive, orthopedic, respiratory, and integumentary disorders. Methods for elimination or reduction of these mortality factors are also discussed. PMID- 9220133 TI - Liver torsion and associated bacterial peritonitis in a dog. AB - A Malamute was examined for acute abdominal pain and collapse. Radiographs of the abdomen showed a pneumoabdomen, and a lucent mass in the region of the liver. Abdominocentesis yielded an inflammatory exudate with bacteria compatible with a Bacillus or a Clostridium sp. A quadrate lobe torsion was found at postmortem. PMID- 9220132 TI - Streptococcus agalactiae mastitis: a review. AB - Streptococcus agalactiae continues to be a major cause of subclinical mastitis in dairy cattle and a source of economic loss for the industry. Veterinarians are often asked to provide information on herd level control and eradication of S. agalactiae mastitis. This review collects and collates relevant publications on the subject. The literature search was conducted in 1993 on the Agricola database. Articles related to S. agalactiae epidemiology, pathogen identification techniques, milk quality consequences, and control, prevention, and therapy were included. Streptococcus agalactiae is an oblique parasite of the bovine mammary gland and is susceptible to treatment with a variety of antibiotics. Despite this fact, where state or provincial census data are available, herd prevalence levels range from 11% (Alberta, 1991) to 47% (Vermont, 1985). Infection with S. agalactiae is associated with elevated somatic cell count and total bacteria count and a decrease in the quantity and quality of milk products produced. Bulk tank milk culture has, using traditional milk culture techniques, had a low sensitivity for identifying S. agalactiae at the herd level. New culture methods, using selective media and large inocula, have substantially improved the sensitivity of bulk tank culture. Efficacy of therapy on individual cows remains high. Protocols for therapy of all infected animals in a herd are generally successful in eradicating the pathogen from the herd, especially if they are followed up with good udder hygiene techniques. PMID- 9220135 TI - Dysfibrinogenemia or afibrinogenemia in a Border Leicester lamb. AB - Hereditary fibrinogen deficiency is a rare condition in all species. Measurement of plasma fibrinogen should indicate low levels. Specific factor assays and pedigree analysis are essential in establishing a definitive diagnosis of the hereditary deficiency. Differentiation between afibrinogenemia, hypofibrinogenemia, and dysfibrinogenemia requires sophisticated techniques and assistance from a specialized laboratory. PMID- 9220134 TI - Leptospirosis in raccoons in Quebec: 2 case reports and seroprevalence in a recreational area. AB - Raccoons may represent a source of leptospires for humans and domestic animals. We describe a case of severe interstitial nephritis associated with the serovar bratislava of Leptospira interrogans (1st report in wildlife), and the seroprevalence to 4 leptospire serovars in a recreational area in Quebec. PMID- 9220137 TI - Strains fall mainly on the transverse plane. PMID- 9220136 TI - Incomplete nasomaxillary dysplasia in a foal. AB - Atresia of the nasal punctum is the most common congenital anomaly for the equine nasolacrimal system. Nasomaxillary dysplasia has not been previously documented in foals, is of unknown etiology, and appears to be a rare condition. Conjunctivomaxillary sinostomy was successful in resolving the epiphora. PMID- 9220138 TI - Extraction of the lower cuspid, with minimal stress. PMID- 9220139 TI - Veterinary applications of the multifunction knife. PMID- 9220140 TI - Analysis of primate major histocompatibility complex (MHC)-DQA1 locus by PCR single strand conformation polymorphism (SSCP). AB - Major histocompatibility complex (Mhc) gene products play an important role in the immune responses against pathogens and autoimmunity, disease resistance and transplantation. Non-human primates (NHPs) are increasingly being utilized as models to test the safety and efficacy of candidate vaccines. Mhc typing of NHPs is an important component of the vaccine trial studies and in the investigations of any associations between Mhc alleles and disease. Routine typing of primate Mhc alleles has been hampered by unavailability of well characterised immunological reagents. In this study, we have used PCR amplification and SSCP for screening polymorphisms in the primate DQA1 locus. Using this technique, 9 African primate species (36 individuals) were analyzed. Ten individuals showed three or four electrophoretic band patterns and the rest two-band patterns indicating this technique can be used to discriminate homozygous and heterozygous individuals prior to DNA sequencing. This method may also be used to screen primates for Mhc-DQA1 allelic polymorphism. However, practical application of this technique for routine typing of primate Mhc-DQA1 alleles depends on availability of adequate nucleotide sequence information. PMID- 9220142 TI - Characterization of a new non-toxic two-chain ribosome-inactivating protein and a structurally-related lectin from rhizomes of dwarf elder (Sambucus ebulus L.). AB - A new N-glycosidase ribosome-inactivating protein (RIP) belonging to the novel family of the nontoxic type 2 RIPs from Sambucaceae has been isolated from rhizomes of dwarf elder (Sambucus ebulus L.) and named ebulin r. Dwarf elder rhizomes also contain a novel monomeric N-Ac-galactosamine-binding lectin that we named SEAII. Ebulin r and SEAII have two isoforms each one, which were readily resolved by ion exchange. Both isoforms of ebulin (ebulins r1 and r2) strongly inhibited protein synthesis in mammalian but not in plant ribosomes by promoting depurination of sensitive ribosomes. Ebulin r and SEAII have apparent molecular masses of 56 and 33.5 kDa, respectively. Ebulins r1 and r2 are composed of two dissimilar subunits (types A-B) of apparent molecular masses of 26 and 30 kDa by disulphide bridges. The rhizome SEAII and the lectins SNA II and SNA III from elder (Sambucus nigra L.) share good amino acid sequence homology. This rhizome ebulin-A chain is more sequence-related to RIP members of cucurbitaceae than to any other plant family. The rhizome ebulin B chain shares a large homology in amino acid sequence with ebulin 1-B chain and SEAII. Anti-ebulin 1 polyclonal antibodies raised in rabbits reacted better with ebulin r1 than with ebulin r2, thus suggesting that both RIP isoforms could have some differences. PMID- 9220141 TI - Analysis by confocal laser scanning microscopy imaging of undilated bile canaliculi F-actin staining in the hepatocytes of human extrahepatic cholestatic liver. AB - Many studies have demonstrated the role of bile canalicular microfilaments in bile secretion and bile flow. It is now admitted that modification of bile canalicular network of microfilaments play a role in dysfunction of bile secretion observed in many cases of cholestasis. This work intends to study F actin, a major component of microfilaments, in human hepatocytes in extrahepatic cholestasis. Normal and extrahepatic cholestatic liver were studied. F-actin was stained with fluorescent phallotoxin and quantified by using confocal laser scanning microscopy and an image analysis method. Mean specific fluorescence (MSF) of bile canaliculi was measured. Since dilated and bile plugged canaliculi were rarely observed in cholestatic liver sections, only undilated bile canaliculi were analysed. Bile canalicular MSF was significantly increased (p < 0.05) in cholestatic hepatocytes (1.3 to 1.7 fold higher than in controls). These data demonstrate a pericanalicular thickening of F-actin microfilaments in human extrahepatic cholestatis, similar to that described in literature in many cases of human intrahepatic and extrahepatic cholestasis cases as well as in experimentally induced cholestasis. However, further studies are needed to understand this increase in F-actin pericanalicular microfilaments in human extrahepatic cholestasis. PMID- 9220143 TI - Use of confocal microscopy to localize the SHBG interaction with human breast cancer cell lines--a comparison with serum albumin interaction. AB - This work has allowed a comparison between the interaction of two principal plasma estradiol-binding proteins, serum albumin and Sex Hormone Binding Globulin (SHBG), with human breast cancer cells in culture (MCF-7 and MDA-MB 231), using a protocol which protects the integrity of cell structure. We showed that serum albumin was highly internalized by cells whereas SHBG interacted essentially at the plasma membrane level, and this whatever the contents of the receptor estrogen cells. If, by its high plasma concentration, serum albumin is internalized in a non-specific way and can thus fit into intracellular traffic, SHBG, by its membrane binding, seems to have a specific action toward target cells. PMID- 9220145 TI - Extracellular ATP induces cytosolic calcium oscillations associated with an increase in intracellular pH in human Chang liver cells. AB - Phosphoinositide (PI) signal transduction pathways are known to be involved in the modulation of several key cellular functions. In this study, ATP is observed to induce calcium transients which appeared to oscillate depending on the ATP concentration. Since higher doses of ATP are known to be toxic to cells, an increase in amplitude could possibly be an inherent cellular protective mechanism. Mobilization of cytosolic calcium from intracellular stores is mediated by the PI second messenger, inositol 1,4,5 trisphosphate. The raised cytosolic calcium was also associated with a rise in intracellular pH in a concentration dependent manner in ATP-treated cells. Intracellular alkalinization is an important parameter in the control of cellular responses. PMID- 9220146 TI - Plasticity of neuro-endocrine-thymus interactions during aging--a minireview. AB - Thymic regrowth and reactivation of thymic endocrine activity may be achieved even in old animals by different endocrinological or nutritional manipulations. In particular: a) intrathymic transplant of pineal gland or treatment with melatonin; b) implantation of a growth hormone secreting tumor cell line or treatment with exogenous growth hormone; c) castration or treatment with exogenous LH-RH; d) treatment with exogenous thyroxine or triiodothyronine, and e) nutritional interventions such as arginine or zinc supplementation. These data strongly support the idea that thymic involution is a phenomenon secondary to age related alterations in neuroendocrine-thymus interactions and that it is the disruption of such interactions in old age which is responsible for most of age associated dysfunctions. With regard to the mechanisms involved in hormone induced thymic reconstitution, it is, at present, difficult to draw any definitive conclusion. The effect of GH, thyroid hormones and LH-RH may be due to the presence on thymic epithelial cells, supposed to produce thymic peptides, of the specific hormone receptors. Melatonin or pineal derived factors may as well act through specific receptors but experimental demonstration is still lacking. The role of zinc, whose turnover is usually reduced in old age, is of quite wide range: from the reactivation of zinc-dependent enzymes, required for both cell proliferation and apoptosis, to the reactivation of thymulin, a zinc-dependent thymic hormone. The role of zinc may be even more crucial. According to recent preliminary data obtained both in animal and in man, it appears that the above reported endocrinological manipulations, capable of restoring thymic activity in old age, may act also by normalizing the altered zinc pool. PMID- 9220147 TI - 1.25-Dihydroxyvitamin D3 receptor is partly colocalized with oxytocin immunoreactivity in neurons of the male rat hypothalamus. AB - With receptor immunocytochemistry, neurons receptive for the steroidhormone 1.25 dihydroxyvitamin D3 have been observed in hypothalamic nuclei. In the present paper we report that a fraction of 1.25-dihydroxyvitamin D3 receptor (VDR) immunoreactive neurons in the hypothalamus of male rats are immunoreactive for oxytocin (OT), suggesting a direct genomic action of this steroid on OT expression. While only 10% of neurons with OT immunofluorescence in the periventricular nucleus contained nuclear VDR immunostaining, up to 50% of the OT neurons in the supraoptic nucleus and 30% in the magnocellular portion of the paraventricular nucleus were VDR positive. VDR immunostaining in the magnocellular nuclei was in many cases confined to the perinuclear cytoplasm. We assume that 1.25-dihydroxyvitamin D3 has effects on hypothalamic peptidergic systems similar to other steroid hormones. PMID- 9220144 TI - Immunochemical localization of the phylogenetically oldest receptor tyrosine kinase: existence in the marine sponge Geodia cydonium. AB - Until now molecular data, elucidating the basis of invertebrate immunity are lacking. Previously both the gene and different cDNAs, coding for the ancestor of metazoan receptor tyrosine kinases (RTK), have been isolated from the marine sponge Geodia cydonium. The sponge RTK shows high polymorphism in the coding as well as in the non-coding parts of the gene. To further elucidate if the sponge RTK might be a molecule involved in self/non-self recognition the intracellular portion of the sponge RTK was expressed in Escherichia coli. The 59 kDa recombinant protein was used to raise monoclonal antibodies (McAb). The McAb recognized three polypeptides of sizes 135, 68 and 26 kDa by Western blotting. The McAb recognized only the plasma membranes of sponge cells as analyzed by immunohisto- and cytochemical studies. Northern blotting analysis revealed that the expression of the RTK gene depends on environmental conditions and on seasonal variations. Based on the high degree of polymorphism and on the immunochemical data we suggest that the RTK in G. cydonium might be involved in sponge immunity. PMID- 9220148 TI - Cellular toxicity of 2,4,5-trichlorophenoxyacetic acid: formation of 2,4,5 trichlorophenoxyacetylcholine. AB - One of the toxic symptoms of 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) is reduction in metabolic rate and subsequent growth retardation. Acetylcholine (ACh) serves as an essential growth factor to facilitate amino acid transport and to promote fetal growth. Hydatidiform mole lacks the capacity for synthesis of ACh, and inhibition of ACh synthesis depresses placental amino acid transport. Therefore, we studied the formation of 2,4,5-acetylcoenzyme A (2,4,5-T-CoA) by acetylcoenzyme A synthase (ACoAS) and the formation of 2,4,5-T-ACh by human placental choline acteyltransferase (ChA) from 2,4,5-T-CoA and choline. In these studies, the widely used analog of 2,4,5-T as an herbicide, 2,4 dichlorophenoxyacetic acid (2,4-D), was also included. These studies have the following results (M +/- S.D.; N,6):1) The enzymatic rates of formation of acetyl CoA, 2,4,5-T-CoA, and 2,4-D-CoA by ACoAS were 32 +/- 4, 23 +/- 3 and 26 +/- 8 nmol/mg protein/5 min., respectively; 2) There were no significant differences in the maximal amounts (nmol/mg protein) of acetyl-CoA (128 +/- 4), 2,4,5-T-CoA (125 +/- 8) and 2,4-D-CoA (96 +/- 6) formed during the reaction period of 50 min.; 3) 14C-2,4-ACh was formed from 14C-2,4-D-CoA and choline by placental-ChA; 4) Low concentrations (EC50 1-2 microM) of synthetic 2,4,5-T-ACh and 2,4-D-ACh decreased the contraction heights of the rat phrenic nerve-hemidiaphragm when the nerve or the muscle was electrically stimulated, and 5) Similar results were obtained with 2,4,6-T-ACh, an analog of 2,4,5-T-ACh. These observations indicate that chlorophenoxyherbicides form false cholinergic messengers in the nerve, muscle and placenta. These false cholinergic messengers can be formed at both muscarinic and nicotinic synaptic sites and also in non-neuronal cells, where ACh plays an important regulatory role as a local hormone, and act as blocking agents. These results will partially explain myotonia, ventricular fibrillation and fetal growth retardation induced by these herbicides. PMID- 9220150 TI - Effect of exogenous growth hormone on in vitro proliferation of thymic lymphocytes from the hypophysectomized rats. AB - Thymus weight and total thymic lymphocyte (TL) number decreased markedly after hypophysectomy (HX). Serum level of insulin-like growth factor-I (IGF-I), one of mediators brought about by growth hormone (GH), reduced considerably after the operation. Exogenous GH stimulation enhanced significantly DNA synthetic activity of TLs from HX group only at lower cell concentrations, less than 5 x 10(5) cell/ml. However, there was not a significant difference in the percentage increase of the TL-DNA synthetic activity with exogenous GH between HX and sham operated (SO) groups. Since GH-responding TLs appeared to be mature cells and to exist in thymus medulla, the results suggest that rate of the responding cells in the thymus medulla of HX animals is similar to that of SO ones. Furthermore, there was not a significant difference in the percentages of rosette-forming cells and non-rosette-forming cells, reflecting nearly the maturation steps of TLs, between groups. It is difficult to explain such an aspect in spite of severe reduction of TLs after hypophysectomy, since the proliferative-responding rate of TLS to exogenous GH stimulation and the constitutive ratio of each maturation step of TLs in the HX group were almost same as those of SO group. The contradictory data found in thymus after hypophysectomy are discussed from the point of view of the compensatory effects of growth hormone-releasing hormone on TLs from the operated animals. PMID- 9220149 TI - Cholinergic markers in transformed trophoblast cells: BeWo and JAr cells. AB - During the first trimester of pregnancy, the human placental trophoblast contains very low levels of choline acetyltransferase (ChA), a specific enzyme for synthesis of acetylcholine (ACh) and a reliable cholinergic marker. There is a relationship between ChA levels and development and maturation of syncytiotrophoblast during the first six months of pregnancy, when ChA levels reach maximum. These observations suggest that ChA and its product, ACh, have significant roles during differentiation of cytotrophoblast into syncytium. Therefore, we have measured ChA levels in malignant trophoblast cultures before and after differentiation. Two pure trophoblast cell lines (BeWo and JAr) are used in these studies. ChA in these cells was determined by a standard radiometric method in which 14C-acetyl groups were transferred from 14C acetylcoenzyme A to choline. Methotrexate (1 microM) was used to differentiate the cells. The following results were obtained: 1) Undifferentiated BeWo cells contained ChA level of 311.0 +/- 8.3 pmoles ACh formed (M +/- S.E., N = 6)/mg protein/10 min. Differentiation decreased ChA level to 213.0 +/- 9.3 pmoles ACh/mg protein/10 min.; b) Undifferentiated JAr cells contained ChA levels of 279.0 +/- 15.0 ACh pmoles formed/mg protein/10 min. This decreased to 166.8 +/- 10.0 upon differentiation; c) Upon differentiation, ChA levels decreased by 31.5 40.2% in BeWo and JAr cell lines. In term human placenta, the ChA levels falls by about 46.0-73% after syncytiotrophoblast is fully developed; d) Chorionic gonadotropin (hCG), which is produced in cytotrophoblast, is a standard tumor marker for chorionic cancer. As the syncytiotrophoblast is formed, production of hCG decreases. Similarly, production of hCG by BeWo and JAr cells decreases upon their differentiation, and e) In hydatidiform mole, which can undergo malignant transformation into choriocarcinoma, there were no significant levels of ChA. These observations suggest that ChA and ACh are necessary for development and maturation of syncytiotrophoblast. PMID- 9220151 TI - Effect of notoginsenoside R1 on the synthesis of components of the fibrinolytic system in cultured smooth muscle cells of human pulmonary artery. AB - We have previously reported that notoginsenoside R1 (NG-R1) increases the synthesis of tissue-type plasminogen activator (t-PA) and decreases plasminogen activator inhibitor-1 (PAI-1) activity in cultured human endothelial cells from different vascular sources. It was the aim of this study to investigate whether the effect of NG-R1 on the synthesis of components of the fibrinolytic system is also operative in another cell type of the blood vessel wall, the smooth muscle cell. Therefore cultured human pulmonary artery smooth muscle cells (HPASMCs) were treated with NG-R1. When the HPASMCs (passage 4 or 5) were conditioned with NG-R1, a dose (0.01-100 micrograms NG-R1/ml for 24 hrs.) dependent increase in t PA an u-PA synthesis was observed, which was significant from 1 microgram NG R1/ml on. t-PA antigen increased from 2.4 +/- 0.1 to 4.7 +/- 0.5 ng/10(5) cells/24 hrs.; u-PA antigen increased from 1.8 +/- 0.1 to 3.0 +/- 0.4 ng/10(5) cells/24 hrs. In contrast no change in PAI-1 antigen synthesis was seen in the conditioned media from NG-R1 treated HPASMCs. On Northern blot analysis of mRNA obtained from NG-R1-stimulated and control HPASMCs NG-R1 induced a significant increases in mRNA levels of t-PA and u-PA (180% and 200% of control value, respectively) at 100 micrograms NG-R1/ml while PAI-1 mRNA decreased slightly. In conclusion our data give evidence that NG-R1 can increase the fibrinolytic potential in cultured HPASMCs in vitro by increasing the production of t-PA and u PA. If operative in vivo this effect of NG-R1 on the fibrinolytic system of SMCs might also contribute to the effect of the Chinese herb drug Panax notoginseng in the treatment of cardiovascular diseases. PMID- 9220152 TI - Quantitative studies on liver fibrosis and alpha-smooth muscle actin expression in heroin abusers. AB - Lobular hepatic fibrosis and the presence of myofibroblasts were studied in heroin abusers, by quantitative automatic image analysis. Nineteen addicts (DA) and thirteen patients having stopped consumption (exDA) were compared to a non addict group (CONTROL). Addicts, all anti-HIV and HBsAg negative, showed increased transaminase levels. Hepatitis C markers were ot available, at the time of biopsy. The surface of the centrolobular fibrosis, measured on picrosirius stained slides, was respectively 1.9 and 3.5 times larger in DA and exDA than in CONTROL (p < 0.0001). Immunolabelling with an alpha-smooth muscle actin antibody (alpha-SMA) revealed stellate cells in a perisinusoidal location, mainly in areas of matrix thickening in the space of Disse. Morphometric analysis of alpha-SMA expression showed significant differences between the three groups of patients, p < 0.0001 (CONTROL: 198.06 +/- 5.59 microns2; DA: 2227.91 +/- 88.02 microns2; exDA: 3469.10 +/- 154.98 microns2). The surface density of collagen and of alpha SMA reactivity was also significantly different between these groups (p < 0.0001). These data strongly suggest that heroin is responsible for an early and progressive centrolobular liver fibrosis, occurring simultaneously with a myofibroblastic response. It might represent a reparative phenomenon arising from a direct vascular injury, leading to an impairment of blood-hepatocyte exchange. PMID- 9220153 TI - Clinical, bacteriological, and serological aspects of Klebsiella infections and their spondylarthropathic sequelae. PMID- 9220155 TI - Development and application of genetic probes for detection of Enterocytozoon bieneusi in formalin-fixed stools and in intestinal biopsy specimens from infected patients. AB - The microsporidium Enterocytozoon bieneusi is closely linked to wasting and diarrhea in a high proportion of individuals with AIDS. However, its relative contribution to disease is uncertain because diagnosis until recently depended on procedures involving endoscopy. A sensitive PCR technique which amplifies a fragment of the small-subunit rRNA gene of E. bieneusi from formalin-fixed stool samples was developed. Of 80 formalin-fixed stool samples collected from 74 Zimbabweans and 6 U.S. patients who were human immunodeficiency virus positive, 50% tested positive for E. bieneusi by PCR, whereas 24% tested positive for E. bieneusi by light microscopy of trichrome-stained fecal smears. In addition, we describe an in situ hybridization technique which detected and identified E. bieneusi as the causative agent in all six intestinal biopsy specimens tested. Both the PCR and in situ hybridization procedures are sensitive diagnostic tools which will complement currently available techniques and enable the differentiation of E. bieneusi from other microsporidia to be made. PMID- 9220154 TI - Immunoblot reactivity of polyclonal and monoclonal antibodies with periplasmic flagellar proteins FlaA1 and FlaB of porcine Serpulina species. AB - The periplasmic-flagellum (PF) proteins of Triton X-100-soluble and Triton X-100 insoluble sodium dodecyl sulfate-treated fractions from reference and field strains of Serpulina hyodysenteriae, Serpulina innocens, and Serpulina pilosicoli were characterized by Western blotting with a rabbit polyclonal antibody (PAb) specific for the 44-kDa PF sheath protein of S. hyodysenteriae (Z. Li, F. Dumas, D. Dubreuil, and M. Jacques, J. Bacteriol. 175:8000-8007, 1993) and a murine monoclonal antibody (MAb), designated 7G2, specific for the PF core FlaB proteins of S. hyodysenteriae. The MAb 7G2 reacted with a conserved epitope present in the 37-, 34-, and 32-kDa PF core FlaB proteins of all Serpulina species. This suggested that the core FlaB proteins are conserved among porcine Serpulina species. An immunoreactive band of approximately 44 kDa was present with all S. hyodysenteriae, S. innocens, and S. pilosicoli strains that were reacted with the PAb. The specificities of the PAb and the MAb for the FlaA1 and FlaB proteins of Serpulina species were confirmed by N-terminal amino acid sequencing of 44- and 37-kDa proteins, respectively, of S. hyodysenteriae and S. pilosicoli. Results from this study provide further evidence that the 44-kDa protein FlaA1 and the 37 , 34-, and 32-kDa FlaB proteins are conserved among porcine Serpulina species. PMID- 9220156 TI - Preparation of a monoclonal antibody specific for Entamoeba dispar and its ability to distinguish E. dispar from E. histolytica. AB - A monoclonal antibody (MAb), MAb ED17 (immunoglobulin G2a [IgG2a]), prepared against trophozoites of Entamoeba dispar SAW1734RclAR cultured monoxenically with Crithidia fasciculata, reacted with 25 of 26 isolates of E. dispar by an indirect fluorescent-antibody test. In contrast, the MAb failed to react with any of 20 isolates of E. histolytica or other enteric protozoan parasites. Western blot (immunoblot) analysis showed that the molecular mass of the E. dispar antigen recognized by the MAb was 160 kDa under reduced conditions. Immunoelectron microscopy revealed that the antigen was mainly located on digested C. fasciculata, but not on undigested organisms. Double staining with a mixture of MAb ED17 and MAb 4G6 (an IgG1 MAb which reacts exclusively with E. histolytica), followed by incubation with a mixture of fluorescein isothiocyanate-labeled anti mouse IgG2a and tetramethylrhodamine isothiocyanate-labeled anti-mouse IgG1 antibodies, simultaneously identified mixed populations of E. dispar and E. histolytica. This method may prove to be useful for the accurate identification of E. dispar and E. histolytica, even in mixed infections. PMID- 9220157 TI - Standardization of an opsonophagocytic assay for the measurement of functional antibody activity against Streptococcus pneumoniae using differentiated HL-60 cells. AB - Host protection against pneumococcal disease i primarily mediated by phagocytosis. We developed and standardized an opsonophagocytic assay using HL-60 cells (human promyelocytic leukemia cells). Fifty-five serum samples were analyzed for the presence of functional antibody against seven pneumococcal serogroups or serotypes (4, 6B, 9V, 14, 18C, 19F, and 23F) by using differentiated HL-60 cells (granulocytes) and peripheral blood leukocytes (PBLs). Six of the 55 serum samples were from unvaccinated adult volunteers, 31 serum samples were from adults who received one dose of the 14-valent or the 23-valent polysaccharide vaccine, and 18 serum samples were from 16-month-old infants who received four doses of an investigational 7-valent polysaccharide-protein conjugate vaccine. The results of an opsonophagocytic assay with HL-60 cells correlated highly with those of an assay with PBLs as effector cells (median r for seven serotypes = 0.87: P < 0.01). Opsonophagocytic titers were compared with the immunoglobulin G antibody concentrations determined by enzyme-linked immunosorbent assay (ELISA). The r values for serogroups or serotypes 4, 6B, 9V, 14, 18C, 19F, and 23F were 0.61, 0.60, 0.67 0.90, 0.61, 0.39, and 0.57, respectively, when HL-60 cells were used as effector cells and 0.56, 0.47, 0.61, 0.90, 0.71, 0.31, and 0.62, respectively, when PBLs were used. The assay requires small amounts of serum (40 microliters per serotype), making this test suitable for assaying infant sera. Culturable cells aid in assay standardization and likely reduce donor-to-donor variability. This standardized assay, in combination with the standardized ELISA, can be used to evaluate current and developing pneumococcal vaccines, in which functional opsonophagocytic antibody activity may correlate with protection against pneumococcal disease. PMID- 9220158 TI - Expression, characterization, and immunoreactivities of a soluble hepatitis E virus putative capsid protein species expressed in insect cells. AB - The hepatitis E virus (HEV) open reading frame-2 (ORF-2) is predicted to encode a 71-kDa putative capsid protein involved in virus particle formation. When insect Spodoptera frugiperda (Sf9) cells were infected with a recombinant baculovirus containing the entire ORF-2 sequence, two types of recombinant proteins were produced; an insoluble protein of 73 kDa and a soluble protein of 62 kDa. The 62 kDa species was shown to be a proteolytic cleavage product of the 73-kDa protein. N-terminal sequence analysis of the 62-kDa protein indicated that it lacked the first 111 amino acids that are present in the full-length 73-kDa protein. A soluble 62-kDa protein was produced without the proteolytic processing by inserting the coding sequence of amino acids 112 to 660 of ORF-2 in a baculovirus expression vector and using the corresponding virus to infect Sf9 cells. The two recombinant 62-kDa proteins made by different mechanisms displayed immunoreactivities very compatible to each other. The 62-kDa proteins obtained by both proteolytic processing and reengineering demonstrated much higher sensitivities in detecting anti-HEV antibodies in human sera than the antigens made from bacteria, as measured by enzyme-linked immunosorbent assay. The data suggest that the soluble 62-kDa protein made from insect cells contains additional epitopes not present in recombinant proteins made from bacteria. Therefore, the 62-kDa protein may be useful for HEV diagnostic improvement and vaccine development. The reengineered construct allows for the consistent large scale production of the soluble 62-kDa protein without proteolytic processing. PMID- 9220159 TI - Immune response to the mannose-sensitive hemagglutinin in patients with cholera due to Vibrio cholerae O1 and O0139. AB - The mannose-sensitive hemagglutinin (MSHA) is a type 4 pilus present in Vibrio cholerae O1 strains of the El Tor biotype, as well as in strains of serogroup O139. It has been shown to be a colonization antigen in animal models. The aim of this study was to investigate systemic and local antibody responses to MSHA in adult patients with cholera due to V. cholerae O1 and O139. Twenty-four of 28 (86%) patients with O1 cholera and 11 of 17 (65%) patients with O139 cholera showed significant increases in MSHA-specific immunoglobulin A (IgA) and IgM antibody-secreting cells (ASCs) 7 days after the onset of disease. However, the magnitude of the ASC response in O1 cholera patients was significantly higher than that in the O139 cholera patients in both IgA-producing (P = 0.015) and IgM producing (P = 0.029) cells. Both groups of patients responded with antibody responses to MSHA in plasma, seroconverting with both IgA (63 to 70% of patients) and IgG (43 to 59% of patients) antibodies. Compared to the MSHA-specific antibody levels determined in healthy controls (n = 10), more than 90% of O1 and O139 cholera patients showed responses to MSHA of both the IgA and the IgG isotypes. About 70% of the patients in both groups also had antibody responses to MSHA in their feces. In summary, we demonstrated that MSHA is immunogenic, giving rise to both systemic and local antibodies in patients with cholera due to both O1 and O139 serogroups. PMID- 9220160 TI - MRP 8/14 as marker for Plasmodium falciparum-induced malaria episodes in individuals in a holoendemic area. AB - Presence of Plasmodium falciparum parasites in the peripheral blood of patients in a holoendemic area does not necessarily show that their illness is due to malaria. The aim of the present project was therefore to look for biological markers related to symptomatology or clinical events during a malaria episode. We focused our work on a complex of heterodimeric calcium-binding proteins secreted by stimulated neutrophils and monocytes, named MIF or myeloid-related proteins (MRP 8/14). In a longitudinal study including 51 adults from Ifakara, Tanzania (84.7% prevalence for P. falciparum in adults during the study), the level of MRP 8/14 in the serum was significantly related to the parasite load (Spearman correlation coefficient, 0.52; P < 0.0001). In the serum from children up to 6 years admitted at a health post the MRP 8/14 levels were closely related to parasitemia but also to fever episodes (Spearman correlation coefficients, 0.96 and 0.736; P < 0.0001 and P < 0.001, respectively). Although not specific to malaria, the measurement of MRP 8/14 could be an additional tool in assessing malaria-related morbidity. PMID- 9220161 TI - T-cell, antibody, and cytokine responses to homologs of the 60-kilodalton heat shock protein in Helicobacter pylori infection. AB - For Helicobacter pylori, the hsp60 heat shock protein encoded by hspB is being considered as a potential candidate for subunit vaccines. We investigated the humoral and cellular responses to H. pylori hsp60 and its cross-reactivity with the homologous Mycobacterium bovis p65 protein and autologous human hsp60 protein. H. pylori-infected persons had significantly higher levels than uninfected persons of serum immunoglobulin G antibodies recognizing H. pylori hsp60, but not M. bovis p65 or human hsp60, as determined by enzyme-linked immunosorbent assay. In contrast, immunoblotting demonstrated cross-reactivity of H. pylori hsp60 with human hsp60. T-cell recognition of H. pylori hsp60 was found in both infected and uninfected subjects, and there was no recognition of human hsp60. T cells from infected and uninfected subjects that had been activated in response to H. pylori hsp60 or M. bovis p65 were phenotypically similar but appeared to secrete different levels of gamma interferon and interleukin-10. These results demonstrate an apparent difference in the epitopes recognized by the T and B cells responding to H. pylori hsp60 in H. pylori-infected persons. In contrast to the T-cell responses, which were highly variable in all subjects and showed no recognition of autologous proteins, a specific B-cell response that may have cross-reactivity to human hsp60 is evident in some infected subjects. PMID- 9220162 TI - Perioperative variation in phagocytic activity against Candida albicans measured by a flow-cytometric assay in cardiovascular-surgery patients. AB - Candidiasis is an opportunistic fungal infection that frequently occurs following modifications of host defenses. Major surgery can be responsible for such alterations, and therefore it increases the risk of fungal infection. The purpose of this study was to evaluate the perioperative impairment of leukocyte function in patients after cardiovascular surgery by measuring the phagocytic activity against Candida albicans by a flow-cytometric method. The average postsurgical decrease in phagocytosis in our patients was 11.4%. By univariate analysis, three factors, all related to antibiotic therapy, were significantly associated with an important decrease in phagocytosis; the use of antimicrobial therapy before surgery, the number of different antibiotics taken, and the length of antibiotic treatment. The results of our study showed that the use of antibiotics in patients undergoing cardiovascular surgery alters the normal phagocytic activity of the host immune system against C. albicans and that flow cytometry is a rapid and simple technique that helps in early identification of patients at high risk for Candida infections. The mechanisms by which surgery and antibiotics decrease phagocytosis remain to be elucidated. PMID- 9220163 TI - Evaluation of a dipstick enzyme-linked immunosorbent assay for detection of antibodies to dengue virus. AB - Accurate serological confirmation of dengue (DEN) infection is difficult, because simple reliable assays for the detection of DEN antibodies are not available. To address this problem, a dipstick enzyme-linked immunosorbent assay (ELISA) was evaluated. The dipstick contained dots of serially diluted DEN 2 antigen. To detect immunoglobulin G (IgG), the dipstick was processed through four reaction cuvettes containing test serum, enhancer, enzyme-conjugated anti-human IgG and IgM antibody, and substrate. Total assay time was 45 min. To detect IgM, the serum was passed through a protein G device to remove IgG. The dipstick was then processed as before, except that the incubation times were longer and enzyme conjugated anti-human IgM was used. The total assay time was 3 h. The dipstick ELISA results were compared with results from microplate ELISA. The IgG dipstick ELISA showed a sensitivity of 95.2% and a specificity of 100% compared to an IgG microplate ELISA with serum samples from 125 individuals living in an area in which DEN is endemic. In tests with 75 serum samples from patients with clinically suspected acute DEN infections, the IgM dipstick ELISA showed a sensitivity of 97.9% and specificity of 100% compared to those of an IgM antibody capture microplate ELISA. These results showed that the dipstick ELISA was a sensitive and specific test for the detection of either DEN IgM or IgG in human serum. The dipstick ELISA was also shown to be useful for detecting seroconversions to DEN IgM or IgG in paired serum samples from 20 patients with virus isolation-confirmed acute DEN infections. PMID- 9220164 TI - Inhibition of Prevotella and Capnocytophaga immunoglobulin A1 proteases by human serum. AB - Oral Prevotella and Capnocytophaga species, regularly isolated from periodontal pockets and associated with extraoral infections, secret specific immunoglobulin A1 (IgA1) proteases cleaving human IgA1 in the hinge region into intact Fab and Fc fragments. To investigate whether these enzymes are subject to inhibition in vivo in humans, we tested 34 sera from periodontally diseased and healthy individuals in an enzyme-linked immunosorbent assay for the presence and titers of inhibition of seven Prevotella and Capnocytophaga proteases. All or nearly all of the sera inhibited the IgA1 protease activity of Prevotella buccae, Prevotella oris, and Prevotella loescheii. A minor proportion of the sera inhibited Prevotella buccalis, Prevotella denticola, and Prevotella melaninogenica IgA1 proteases, while no sera inhibited Capnocytophaga ochracea IgA1 protease. All inhibition titers were low, ranging from 5 to 55, with titer being defined as the reciprocal of the dilution of serum causing 50% inhibition of one defined unit of protease activity. No correlation between periodontal disease status and the presence, absence, or titer of inhibition was observed. The nature of the low titers of inhibition in all sera of the IgA1 proteases of P. buccae, P. oris, and P. loescheii was further examined. In size exclusion chromatography, inhibitory activity corresponded to the peak volume of IgA. Additional inhibition of the P. oris IgA1 protease was found in fractions containing both IgA and IgG. Purification of the IgG fractions of five sera by passage of the sera on a protein G column resulted in recovery of inhibitory IgG antibodies against all three IgA1 proteases, with the highest titer being for the P. oris enzyme. These finding indicate that inhibitory activity is associated with enzyme-neutralizing antibodies. PMID- 9220165 TI - Vaginal formulations of carrageenan protect mice from herpes simplex virus infection. AB - The observations from the present study indicate that vaginal formulations of the sulfated polysaccharide carrageenan are highly effective in protecting mice from herpes simplex virus type 2 (HSV-2) infection. Test formulations were placed in the vaginas of progestin-treated mice prior to inoculation with HSV-2. Infection was determined by the presence of inflammation in the genital region and death. At a dose of virus that infected half of the control animals, 1% solutions of either lambda, kappa, or iota carrageenan prevented infection of almost all of the animals. Concentrations as low as 0.05% protected a large majority of the mice. At a dose of virus that infected all of the control mice, 1% solutions of carrageenans protected 85% of the inoculated mice. Other sulfated polysaccharides were less effective or showed no efficacy in preventing HSV-2 infection. These findings suggest that a vaginal formulation of carrageenan may be effective in blocking sexual transmission of HSV-2 in women. PMID- 9220166 TI - Avidity of immunoglobulin G directed against human cytomegalovirus during primary and secondary infections in immunocompetent and immunocompromised subjects. AB - Diagnosis of primary human cytomegalovirus (HCMV) infection is accomplished exclusively by serologic testing. Among the possible methods, the determination of immunoglobulin G (IgG) avidity is one of the least explored. In this work, we used a commercially available kit to test anti-HCMV IgG avidity in 336 serum samples from pregnant women and transplant recipients undergoing virologically proven HCMV primary or nonprimary infections and from latently infected blood donors. Our results demonstrate that the anti-HCMV IgG avidity test differentiates primary from nonprimary HCMV infections in both pregnant women and solid organ transplant recipients. In fact, 88.6% of primary infections and no secondary infections showed low-avidity IgG to HCMV. In particular, low IgG avidity is a marker of primary infection for 18 to 20 weeks after onset of symptoms in both immunocompromised and immunocompetent subjects. PMID- 9220167 TI - Immunological markers of disease progression in patients infected with the human immunodeficiency virus. AB - Identification of inexpensive and technically simple immunological tests useful in predicting the progression to AIDS in human immunodeficiency virus (HIV) infected patients would be especially welcome in developing countries, in which 80% of HIV-infected patients reside and health budgets are low. In the current study, we evaluated CD4+ and total lymphocyte counts and the concentrations in serum of beta 2-microglobulin, p24 antigen, and immunoglobulin A (IgA) as predictors of disease progression in 74 Panamanian HIV-positive patients and 50 HIV-negative healthy individuals. Total lymphocyte and CD4(+)-cell counts for AIDS patients (1,451 +/- 811 cells/microliters, P < 0.001, and 238 +/- 392 cells/microliters, P < 0.0001, respectively and asymptomatic patients (2,393 +/- 664 cells/microliters, P > 0.05, and 784 +/- 475 cells/microliters, P < 0.001, respectively) were lower than those observed for healthy subjects (2,596 +/- 631 cells/microliters and 1,120 +/- 296 cells/microliters, respectively). The levels of beta 2-microglobulin and IgA in serum were significantly elevated in patients with AIDS (5.7 +/- 3.6mg/liter, P < 0.001, and 541 +/- 265 mg/dl, P < 0.0002, respectively) and asymptomatic infected subjects (3.4 +/- 2.1 mg/liter, P = 0.001, and 436 +/- 216 mg/dl, P < 0.0001, respectively) compared with the levels in healthy subjects (2.2 +/- 0.7 mg/liter and 204 +/- 113 mg/dl, respectively). Nonstatistically significant differences (P > 0.05) for concentrations of p24 antigen between asymptomatic infected patients (29 +/- 13 pg/ml) and AIDS patients (40 +/- 23 pg/ml) were observed. Total lymphocyte counts of 1,750 cells/microliters or less, CD4 counts of 200 cells/microliters or less, beta 2 microglobulin concentrations in serum of 4 mg/liter or higher, concentrations of IgA in serum of 450 mg/dl or higher, and the presence in serum of p24 antigen were correlated with elevated risks for developing AIDS. Monitoring both total lymphocytes and beta 2-microglobulin identified 91% of the AIDS patients; these assays may allow reductions in the annual number of CD4(+)-cell evaluations and the costs associated with monitoring both total lymphocytes and beta 2 microglobulin identified 91% of the AIDS patients; these assays may allow reductions in the annual number of CD4(+)-cell evaluations and the costs associated with monitoring the immune status of HIV-positive patients. PMID- 9220168 TI - Detection of serum antibodies to CagA and VacA and of serum neutralizing activity for vacuolating cytotoxin in patients with Helicobacter pylori-induced gastritis. AB - Thirty patients with dyspepsia, with histological diagnosis of gastritis, and with endoscopic diagnosis of peptic ulcer disease (PUD) (n = 13) or nonulcer dyspepsia (NUD) (n = 17) were admitted to the study. Helicobacter pylori vacuolating cytotoxin-producing strains (Tox+) were isolated from 14 (46.7%) patients, whereas non-cytotoxin-producing (Tox-) H. pylori strains were isolated from the remaining patients. Of 30 patients studied, 20 (66.7%) had serum cytotoxin neutralizing activity in vitro. Fourteen patients with Tox+ H. pylori strains showed serum cytotoxin neutralizing activity and serum immunoglobulin G (IgG) and IgA antibodies reactive with both 87-kDa H. pylori vacuolating cytotoxin (VacA) and 128-kDa cytotoxin-associated gene product (CagA) by immunoblotting using native enriched preparations of VacA and CagA proteins from H. pylori culture supernatants as the antigens. A 94-kDa antigen cross-reacting with the 87-kDa VacA protein could be demonstrated in culture supernatant with immune sera from humans and animals. All patients (n = 10) lacking serum neutralizing activity were also negative for IgG or IgA against VacA antigen, whereas 6 of the 10 patients showed IgG serum antibody responses against CagA antigen. The prevalence of antibodies to VacA and CagA antigens was significantly (P < 0.001) higher in patients with gastritis (20 and 26 patients for VacA and CagA, respectively, of 30 patients) than in H. pylori culture-negative controls (0 of 27 for both VacA and CagA) and in randomly selected blood donors (17 and 21 for VacA and CagA, respectively, of 120 subjects). All patients with PUD had antibodies to CagA, whereas 13 of 17 (76.5%) patients with NUD had anti-CagA antibodies. Serum IgG antibodies to VacA were present in 9 (69.2%) patients with PUD of 13 patients and in 11 (64.7%) patients with NUD of 17 patients. Anti-CagA antibodies seemed to correlate better with PUD than anti-VacA antibodies. PMID- 9220169 TI - Search for cytomegalovirus-specific immunoglobulin M: comparison between a new western blot, conventional western blot, and nine commercially available assays. AB - We tested 101 serum samples obtained from pregnant women for the presence of human cytomegalovirus (HCMV)-specific IgM with nine different commercially available kits and with two Western blotting (WB) tests previously developed in our laboratory. The conventional WB test contains viral structural proteins separated on a gel from purified HCMV particles and transferred to nitrocellulose. The new WB test contains viral structural proteins and three recombinant proteins which contain the significant immunogenic portions of pp150 (ppUL32), pp52 (ppUL44), and p130 (pUL57). The results obtained indicate that the new WB test combines high specificity (92.5%) with high sensitivity (95.0%), characteristics that, in combination, have not been obtained with any of the other tests. PMID- 9220170 TI - Production of polyclonal antibodies to feline tumor necrosis factor. AB - Two 13-amino-acid peptides were synthesized based on the putative feline tumor necrosis factor (FeTNF) sequence. The synthesized peptides were conjugated to keyhole limpet hemocyanin, emulsified in complete Freund's adjuvant, and injected into rabbits. The gene for FeTNF was cloned into the FLAG (International Biotechnologies Inc. [IBI], Kodak, New Haven, Conn.) fusion protein expression vector. The expressed fusion protein was purified by using the M-1 anti-FLAG octapeptide monoclonal antibody (IBI, Kodak). The fusion protein was emulsified in complete Freund's adjuvant and injected into chickens. The immune sera generated to the synthetic peptides and the fusion protein recognized the recombinant FeTNF fusion protein on Western or dot blot assay. The preimmune and immune sera were incubated with naturally occurring FeTNF (supernatants from lipopolysaccharide-stimulated cultured feline peritoneal exudate or peripheral mononuclear cells). The antibody raised to the recombinant FeTNF fusion protein and N-terminal synthetic peptide neutralized bioactivity of native FeTNF and recombinant human TNF. Preimmune sera did not have any neutralizing activity. The polyclonal antibodies were not specific for FeTNF, since both porcine and human recombinant TNF were neutralized by the fusion protein antibodies. The synthetic peptide antibodies recognized recombinant feline and equine TNF on a Western blot. PMID- 9220171 TI - Time to earliest peak serum antibody response to influenza vaccine in the elderly. AB - The earliest time at which serum antibody levels peak following administration of an influenza virus vaccine in elderly persons is not clearly defined. We compared the time intervals of 1 and 2 weeks after vaccination. A commercial trivalent vaccine containing the hemagglutinins of influenza viruses A/Texas/36/91 (H1N1), A/Johannesburg/33/94 (H3N2), and B/Harbin/7/94 was used. The hemagglutination inhibition (HAI) antibody titers at 1 week after vaccination were significantly lower than the HAI titers at 2 weeks postvaccination for all three vaccine components. PMID- 9220173 TI - Measuring soluble adhesion molecules. PMID- 9220172 TI - Presence of a neutralizing domain in isolates of rubella virus in Cordoba, Argentina. AB - We studied the presence of a neutralizing epitope of rubella virus (RV) in locally circulating strains in Cordoba, Argentina, using binding by the monoclonal antibody (MAb) H3. This epitope is contained in a sequence of the E1 glycoprotein (E1208-239) represented by the synthetic peptide SP15. H3 MAb showed specific binding to SP15 by enzyme-linked immunosorbent assay (ELISA). One wild type postnatal isolate, four clones derived from this isolate, and one congenital isolate were reactive with H3 by ELISA. These results suggest that the region of RV represented by SP15 is a domain present in locally circulating strains. PMID- 9220174 TI - Abnormalities of the S-T segment--Part II. PMID- 9220175 TI - Ischemic preconditioning: clinical relevance and investigative studies. AB - Experimental animal studies have shown that repetitive brief coronary occlusions render the heart resistant to myocardial infarction from subsequent, more prolonged, coronary occlusions. This phenomenon in animal models has been called ischemic preconditioning. In a number of clinical scenarios, the second in a series of ischemic episodes appears to be less severe than the first, suggesting that ischemic preconditioning also occurs in humans. If the mediator of preconditioning could be identified, it is conceivable that this agent could be administered to patients with coronary artery disease as a myocardial protectant. However, the definite clinical relevance of this interesting experimental finding remains unknown. Unlike the case in animal models subjected to an abrupt occlusion, preconditioning is difficult to study in the clinical setting. This article reviews the findings and limitations of the relevant clinical studies looking for ischemic preconditioning in humans. PMID- 9220176 TI - Amiodarone: the expanding antiarrhythmic role and how to follow a patient on chronic therapy. AB - Amiodarone was introduced as an antiarrhythmic compound in the early 1970s and was approved in the U.S. for the treatment of refractory ventricular arrhythmias in late 1984. Since that time the drug has become the most widely studied antiarrhythmic compound with expanding potential indications, including maintaining stability of sinus rhythm, secondary prevention in the survivors of myocardial infarction, and prolongation of survival in certain subsets of patients with congestive heart failure. Intravenous amiodarone was introduced in the U.S. in 1995 for the control of recurrent destabilizing ventricular tachycardia or ventricular fibrillation resistant to conventional therapy. The level of comfort in its use has risen considerably in the recent past. This has stemmed from the reasonably decisive evidence that class I agents increase mortality in patients with structural heart disease. In contrast, amiodarone either reduces mortality or its effect is neutral; this is consistent with its low to negligible proarrhythmic actions. The drug does not aggravate heart failure and it may even increase left ventricular ejection fraction and improve exercise capacity. Above all, it is becoming increasingly evident from wider experience and from controlled clinical trials that the side-effect profile of the drug is not as compelling an issue as it appeared to be when first used in much higher doses. Therefore, the overall objective of amiodarone therapy is to use the lowest dose that produces a defined therapeutic end point without causing serious side effects. Careful clinical surveillance in conjunction with monitoring of certain laboratory parameters and indices of efficacy at regular intervals permits the drug to be used effectively in a large number of patients who fail to respond to, or are intolerant of other antiarrhythmic compounds. Many experienced clinicians have begun to consider the use of amiodarone as first-line therapy in certain disorders of rhythm, especially in patients with severely compromised ventricular function. PMID- 9220177 TI - Use of cardiac troponin T rapid assay in the diagnosis of a myocardial injury secondary to electrical cardioversion. AB - BACKGROUND AND HYPOTHESIS: This study was carried out to determine whether cardiac troponin T test in rapid assay gives positive results in patients previously submitted to cardioversion or electrical defibrillation. METHODS: Forty patients with supraventricular tachyarrhythmias lasting no more than 2 days were treated with electrical cardioversion. The total creatine phosphokinase (CPK)-MB isoenzyme and troponin T in rapid assay were measured at baseline and at 6, 12, and 24 h thereafter. RESULTS: Total CPK baseline levels were normal in all cases; within 4 h, the serum CPK levels increased by 98%, at 6 h by 111.5%, at 12 h by 168%, and at 24 h by 225% (p > 0.01). The CPK-MB isoenzyme showed no percentage increase of total CPK higher than 5%, measured at 6, 12, and 24 h after the shock, independent of the number of attempts of cardioversion. The troponin T test was also negative in all cases at baseline and at 6, 12, and 24 h after cardioversion. CONCLUSION: We conclude that the absence of elevations in CPK-MB levels and cardiac troponin T levels matched clinical and electrocardiographic results showing absence of myocardial damage after electrical cardioversion. PMID- 9220178 TI - The prognostic significance of angina pectoris experienced during the first month following acute myocardial infarction. AB - BACKGROUND: Angina pectoris accompanied by transient ST-segment changes during the in-hospital phase of acute myocardial infarction (AMI) is a well established marker of subsequent cardiac death and reinfarction. HYPOTHESIS: This study was undertaken to record the prognostic significance of angina pectoris experienced during the first month following discharge from AMI. METHODS: In all, 803 patients included in the placebo arm of the Danish Verapamil Infarction Trial II were followed up for 18 months in 20 coronary care units in Denmark. The patients were randomized to placebo and were still on study treatment 1 month after discharge. Of these patients, 311 (39%) reported chest pain during the first month following discharge. RESULTS: Patients with angina pectoris had a significantly increased risk of reinfarction [hazard 1.71; 95%-confidence limit (CL): 1.09, 2.69] and increased mortality risk which, however, only reached borderline statistical significance (hazard 1.52; 95%-CL: 0.96, 2.40). When patients were subdivided according to both angina pectoris and heart failure, those with one or both of these risk markers had significantly increased mortality (p 0.03) and reinfarction (p 0.02) rates compared with patients free of both angina pectoris and heart failure. CONCLUSION: Patients with postinfarction angina pectoris have a significantly increased morbidity risk. PMID- 9220179 TI - The benefit of low-dose dopamine during vigorous diuresis for congestive heart failure associated with renal insufficiency: does it protect renal function? AB - BACKGROUND: Low-dose dopamine, a renal vasodilator, has been used empirically to improve renal function or outcome in critically ill patients with oliguria or acute renal failure. HYPOTHESIS: This study was designed to investigate the efficacy of low-dose dopamine (2 micrograms/kg/min) as a renal-protective agent during vigorous diuresis for congestive heart failure (CHF) associated with mild or moderate renal insufficiency. METHODS: Of 20 study patients (mean age 74.3 +/- 15 years) with severe CHF, 10 (Group A) were randomized to a treatment strategy of intravenous bumetanide (1 mg b.i.d.) alone and another 10 (Group B) to low dose dopamine and a similar diuretic regimen for a duration of 5 days or less if clinical edema remitted. RESULTS: Group B patients showed a significant improvement in renal function and urinary output: serum blood urea nitrogen 48.9 +/- 10.3 to 32.1 +/- 14.4 mg/dl (p < 0.05); serum creatinine 1.97 +/- 0.24 to 1.49 +/- 0.39 mg/dl (p < 0.05); creatinine clearance 35.6 +/- 11.6 to 48.8 +/- 12.3 ml/min (p < 0.05); and indexed urinary output 0.56 +/- 0.16 to 2.02 +/- 0.72 ml/kg/h (p < 0.05). Group A patients showed a significant increase in urinary output but nonsignificant renal functional deterioration. CONCLUSION: The renal protective effect of low-dose dopamine in the setting of CHF and vigorous diuresis is supported by this study. PMID- 9220180 TI - Circadian rhythmic fractal scaling of heart rate variability in health and coronary artery disease. AB - BACKGROUND: In clinical cardiology, heart rate variability is a putative index of autonomic cardiovascular function. Signs of reduced vagal activity are not only associated with an enhanced risk of sudden cardiac death, but such impaired heart rate variability became a new predictor of sudden cardiac death and other mortality in patients with a variety of diseased states. HYPOTHESIS: It is postulated (1) that the time structure (chronome) of heart rate variability in clinical health includes a circadian rhythm and deterministic chaos, the latter gauged by the correlation dimensions of RR intervals; and (2) that this chronome is altered in patients with coronary artery disease (CAD). METHODS: From 24-h Holter records of 11 healthy controls and 10 patients with CAD, 500-s sections around 02:00, 06:00, 10:00, 14:00, 18:00 and 22:00 hours were analyzed for smoothed RR intervals sampled at 4 Hz. Correlation integrals were estimated for embedding dimensions from 1 to 20 with a 1.0-s time lag, using an algorithm modified from Grassberger and Procaccia. The Wilcoxon signed-rank test compares circadian end points assessed by cosinor between the CAD patients and age-matched controls. RESULTS: A circadian rhythm characterizes the correlation dimension of healthy subjects peaking during the night (p < 0.005). Patients with CAD have a lowered correlation dimension (p < 0.05) and an altered circadian variation which requires the consideration of an approximately 12-h (circasemidian) component. CONCLUSION: The results demonstrate the sensitivity of circadian rhythms for the detection of disease. A partial 24- to 12-h (circadian-to-circasemidian) frequency multiplication (or partial variance transposition) in CAD of the correlation dimension, apart from being a potential clue to the etiology of the disease, adds a new feature to a chronocardiology combining, with the fractal scaling, an assessment of circadian and circasemidian components as measures of predictable variability to be tested for use in diagnosis, prognosis, and as putative guides to treatment timing. PMID- 9220181 TI - Atrial fibrillation: a review of mechanism, etiology, and therapy. AB - The prevalence of elderly individuals in the populations of developed countries is increasing rapidly, and atrial fibrillation (AF) is quite common in these elderly patients: currently, 11% of the U.S. population is between the ages of 65 and 85 years; 70% of people with AF are between the ages of 65 and 85 years. AF causes symptoms secondary to hemodynamic derangements that are the result of increased ventricular response and loss of atrial booster function. AF can lead to reversible impairment of left ventricular function, cardiac chamber dilatation, clinical heart failure, and thromboembolic events. AF requires treatment in order to prevent these potential complications. Type Ia, Ic, and III antiarrhythmics are capable of converting AF to normal sinus rhythm (NSR). Amiodarone has the greatest efficacy and safety for converting AF and maintaining NSR while digoxin and verapamil are ineffective in restoring NSR. Quinidine, flecainide, disopyramide, and sotalol have also been shown to maintain NSR after conversion of AF. Proarrhythmia is a definite concern with the latter four agents. Alternative therapy for AF includes anticoagulation with warfarin or aspirin for the prevention of thromboembolic events, and a variety of agents to control the ventricular response. All medications used to treat AF carry significant risks in the elderly, whether from proarrhythmia, overdosing because of compliance errors, or hemorrhage secondary to anticoagulation. Treatment of AF must be based on a careful risk-benefit evaluation. The physician must know the capability of the particular patient as well as drug mechanisms and effects in the elderly. The decision to convert patients from AF to NSR or to leave the patient in AF and control the ventricular response represents a complex intellectual challenge. Factors favoring one or the other of these two clinical strategies are discussed. Multicenter clinical trials, for example, the Atrial Fibrillation Follow-up Investigation Rhythm Management (AFFIRM) trial, are currently underway to assess various clinical strategies for maintenance of NSR following conversion from AF. Amiodarone is one of the drugs under investigation. PMID- 9220182 TI - Comparison of the histopathology of culprit lesions in chronic stable angina, unstable angina, and myocardial infarction. AB - BACKGROUND: The etiology of unstable angina (UA) and myocardial infarction (MI) both involve rupture of an atherosclerotic plaque in a coronary artery. It has been suggested that the two syndromes differ because MI results if a red occlusive permanent thrombus occurs and UA occurs only if a nonocclusive platelet (white) thrombus occurs. HYPOTHESIS: The purpose of this study was to determine the differences between coronary lesion pathology in MI and UA and compare them with lesions of chronic stable angina (CSA). METHODS: We reviewed the pathologic specimens of culprit lesions obtained by directional coronary atherectomy in 27 patients with MI, 29 patients with UA, and 16 patients with CSA. RESULTS: The incidence of ruptured plaque was high and identical in patients with MI (77.8%), and UA (75.8%), and significantly lower in patients with CSA (25.0%) (p < 0.001). Similarly, the incidence of red thrombus was the same in MI (92.6%) and UA (82.7%), and significantly less in CSA (p < 0.001). CONCLUSIONS: The underlying pathophysiology of both UA and MI appears to be the same, with red thrombus playing an important role in both syndromes. The only difference is in the degree of occlusiveness of the red thrombus on the ruptured plaque and whether the occlusion is transient (UA) or persistent (MI). The balance between thrombosis and endogenous clot lysis determines which syndrome occurs. Lytic therapy is not effective in UA, probably because the clot is not occlusive or because endogenous lysis has already achieved the degree of coronary opening that eventuates from tissue plasminogen activator or streptokinase administration. Prompt catheterization and revascularization may be as indicated in patients with MI if there remains viable myocardium as in patients with UA. PMID- 9220183 TI - Vasospastic angina induced by nonsteroidal anti-inflammatory drugs. AB - We report two cases of vasospastic angina associated with anaphylactic reaction caused by nonsteroidal anti-inflammatory drugs (NSAIDs). Both patients exhibited anaphylactic manifestations, such as general rash and urticaria, along with angina pectoris with electrocardiographic ST-segment elevations after suppository administration of diclofenac sodium or indomethacin, the most commonly used NSAIDs. Although these patients had normal coronary arteriograms, intracoronary administration of ergonovine or acetylcholine provoked diffuse coronary artery spasms accompanied by chest pain and ischemic ST-segment changes. It is therefore suggested that an allergic mechanism may be involved as a causative factor of the coronary artery spasm induced by NSAIDs. PMID- 9220184 TI - Post-traumatic ventricular septal defect following coronary bypass surgery. AB - A 60-year-old patient underwent triple coronary artery bypass grafting following an inferoseptal myocardial infarction and early onset of exertional angina. Four years later he was involved in a car accident during which he sustained an abdominal and thoracic trauma. Approximately 1 month after discharge, a ventricular septal defect was diagnosed by two-dimensional Doppler echocardiography with patency of all grafts at coronary angiography. Closure of the septal defect was successfully accomplished through a right atrial approach. Rupture of the ventricular septum following blunt chest trauma in a patient with previous myocardial revascularization has not been previously reported. PMID- 9220185 TI - Propafenone-induced drug fever in the absence of agranulocytosis. AB - Propafenone is an antiarrhythmic drug used in the treatment of life-threatening ventricular tachyarrhythmias. Adverse reactions necessitating discontinuation of the medication are common. Propafenone-induced drug fever has not been definitively proven. We present a case report of drug fever secondary to propafenone, confirmed with rechallenge. PMID- 9220187 TI - Hypertrophic obstructive cardiomyopathy: alternative therapeutic options. PMID- 9220186 TI - John Eric Erichsen. PMID- 9220188 TI - The autosomal dominant syndrome with congenital stapes ankylosis, broad thumbs and hyperopia. AB - A family is reported with conductive hearing loss, hyperopia, broad thumbs and broad first toes. The family resembles a previous reported family (Teunissen B, Cremers CWRJ (1990) Laryngoscope 100: 380-384) but additionally all affected members have a typical face. Overlap of the Teunissen and Cremers syndrome with the facio-audio-symphalangism syndrome and proximal symphalangism is discussed. PMID- 9220189 TI - Non-haematological traits associated with congenital dyserythropoietic anaemia type 1: a new entity emerging. AB - Dysmorphic features in three sibs with congenital dyserythropoietic anaemia type 1 are described. These findings include growth retardation/short stature, congenital ptosis, abnormal tarsal bones, metatarsal duplication/hypoplasia, nail/phalangeal hypoplasia of fingers and toes, Madelung deformity, syndactyly of toes, and hallux valgus. The patients also showed a very low mitotic index of their peripheral blood lymphocyte cultures. Phenotypic heterogeneity was elicited amongst the three Bedouin sibs. The present report confirms the association between a subset of congenital dyserythropoietic anaemia type 1 and a specific form of distal limb anomalies and suggests that other traits, congenital ptosis and low mitotic index, could represent part of the syndrome profile. PMID- 9220190 TI - Exclusion of the p75 neurotrophin receptor gene as a candidate gene for Meckel syndrome. AB - Nerve growth factor receptor p75 (NGFR) gene was investigated as a potential candidate gene in Meckel syndrome (MKS) because of its important role in embryonic development, chromosomal localization adjacent to the MKS locus and Meckel syndrome-resembling findings in knock-out mice phenotype. The sequence analysis of the coding region of the gene revealed one polymorphism but no potential disease mutation. Physical mapping of the critical chromosomal region finally showed that the NGFR gene lies outside the MKS locus. PMID- 9220191 TI - A probable case of Wiedemann-Rautenstrauch syndrome or neonatal progeroid syndrome and review of the literature. AB - A boy with features suggesting the diagnosis of Wiedemann-Rautenstrauch syndrome (WRS) or neonatal progeroid syndrome is presented. Abnormal findings included a generalized virtual absence of subcutaneous fat, sparse scalp hair, prominence of veins and muscles, a large and persistent anterior fontanelle and facial dysmorphism (triangular aged face, prominent eyes and scalp veins). Until now, only 13 cases (including one prenatal diagnosis) of this syndrome have been described. Since the borderlines of this syndrome are not very exact, we reviewed the previous reports in order to further delineate this rare syndrome. PMID- 9220192 TI - A syndrome of leukonychia totalis and multiple sebaceous cysts. AB - We report on a family with leukonychia totalis, koilonychia and multiple sebaceous cysts segregating as an autosomal dominant disorder. This condition has only previously been described in two kindreds, and this report documents the natural course of the disorder and provides further evidence for pancreatitis being an associated syndromic feature. PMID- 9220193 TI - The syndrome of hypoparathyroidism, severe growth failure, developmental delay and distinctive facies. AB - We report a child from a highly inbred Omani family with hypoparathyroidism, growth failure, developmental delay and a distinctive facial appearance. Thirty cases with this syndrome have been previously reported; 22 came from the Arab Gulf Countries and eight were Arabs living in Israel. These cases are reviewed. PMID- 9220195 TI - Fronto-facio-nasal dysplasia. AB - Fronto-facio-nasal dysplasia is a rare cause of facial clefts. The syndrome is characterized by paramedian facial clefts which involve the nose and palpebral fissures resulting in defects of the alae nasi and blepharophimosis, lagophthalmos, and S-shaped palpebral fissures. In addition affected children have ocular malformations such as epibulbar dermoids and colobomata of the iris or optic disk and may have a posterior encephalocele; these features distinguish this condition from fronto-nasal dysplasia and early amnion rupture sequence. We describe a child with unilateral features. Unilateral craniofacial clefts are usually assumed to have a low recurrence risk. However, fronto-facio-nasal dysplasia is an autosomal recessive condition and must be considered in any child with paramedian facial clefts. PMID- 9220194 TI - Ptosis, down-slanting palpebral fissures, hypertelorism, seizures and mental retardation: a possible new MCA/MR syndrome. AB - We present a family with six children of first cousin parents, in which three present with microcephaly, hypertelorism, down-slanting palpebral fissures, ptosis, a broad nasal tip, a short webbed neck, mental retardation and seizures. Two differential diagnosis, the Noonan and the Baraitser-Winter syndrome are discussed. The possibility of the description of a new MCA/MR syndrome is raised. PMID- 9220196 TI - Oculoauriculofrontonasal spectrum in an adult Brazilian male. AB - We report an adult male patient born to normal and non-consanguineous parents with midline craniofacial defects associated with branchial arch anomalies. This combination of signs belongs to the oculoauriculofrontonasal spectrum. We compare our case with those previously reported who had similar findings and discuss clinical aspects of the oculoauriculofrontonasal spectrum and frontofacionasal dysplasia. PMID- 9220197 TI - Congenital diaphragmatic hernia and ipsilateral limb reduction defect: a new case, long-term follow-up and review of the literature. AB - There have been a small number of documented cases of isolated congenital diaphragmatic hernia and ipsilateral limb defects. Early cervical neural crest injury has been postulated as the mechanism behind the coexistence of these two defects. We present a case of left-sided congenital diaphragmatic hernia and ipsilateral radial ray defect consisting of thumb hypoplasia and absent radius. Our patient is an adult who presented for reproductive counselling providing an opportunity for long-term follow-up. PMID- 9220198 TI - Smith-Lemli-Opitz syndrome: further delineation of the phenotype. PMID- 9220199 TI - Focal dermal hypoplasia (Goltz syndrome) presenting as a severe fetal malformation syndrome. AB - A fetal malformation syndrome comprising growth retardation, anophthalmia, bilateral diaphragmatic herniae, bifid lower leg, syndactyly of the fingers, malrotation of the colon, hypoplastic kidneys and total anomalous pulmonary venous drainage is described in a female fetus from a consanguineous relationship. Differential diagnosis is discussed and it is suggested that this case represents an unusually severe form of Goltz syndrome. PMID- 9220200 TI - Severe malformation of one foot from amniocentesis needle injury. AB - An 11-year-old boy is described who was born with a poorly developed right foot. At 16 weeks gestation his mother had had amniocentesis without direct ultrasound guidance. After the insertion of the amniocentesis needle, she felt strong abdominal resistance, which disappeared with slight withdrawal of the needle. Then, real-time ultrasound was used to determine the position of the needle, which was reported to be up against the feet. Nine days later deep purple amniotic fluid was removed by amniocentesis. At birth there was a scab over an oozing hole in the lateral aspect of his right 'foot', a poorly formed structure with five digit-like structures at the tip. It seems most likely that the deformity was caused by tissue injury from needle puncture at 16 weeks gestation. PMID- 9220201 TI - Oculo-facio-cardio-dental (OFCD) syndrome. PMID- 9220202 TI - Polysyndactyly and trigonocephaly with partial agenesis of corpus callosum: an example of the variable clinical spectrum of the Acrocallosal syndrome? PMID- 9220203 TI - Serpentine fibula syndrome: a variant clinical presentation of Hajdu-Cheney syndrome? PMID- 9220204 TI - Recent advances in the pharmacotherapy of epilepsy. AB - The therapeutic options for the treatment of epilepsy have expanded during the 1990s. Since 1993, four novel agents (felbamate, gabapentin, lamotrigine, and topiramate) have been approved by the US Food and Drug Administration, primarily for adjunctive treatment of partial seizures. In addition, a water-soluble pro drug of phenytoin, fosphenytoin, and a sustained-release preparation of carbamazepine have been introduced. The novel anticonvulsants represent a potential improvement for patients whose seizures are incompletely controlled or who experience significant adverse effects with older anticonvulsants. Felbamate, lamotrigine, and topiramate appear to have a broad spectrum of action in seizure control, but felbamate use is limited by the potential for serious adverse effects. Gabapentin, lamotrigine, and topiramate are all well tolerated. Gabapentin has no known drug interactions, whereas lamotrigine and topiramate have limited interactions compared with older agents. The sustained-release preparation of carbamazepine may decrease the incidence of adverse effects and increase patient compliance. Fosphenytoin offers a safer method for intravenous administration of phenytoin and the added flexibility of intramuscular administration. Taken together, these recent advances in treatment may bring about improved efficacy and decreased adverse effects for many patients with epilepsy. PMID- 9220205 TI - Androgen and estrogen-androgen hormone replacement therapy: a review of the safety literature, 1941 to 1996. AB - The endocrine physiology of the climacteric supports a rationale for the concomitant replacement of androgen and estrogen following menopause. Clinical and research experience with estrogen-androgen hormone replacement therapy, as well as androgen-only therapy, suggests that the health benefit offered by androgen replacement exceeds the potential risk when treatment is properly managed. In this review, we concentrate on the effects of oral alkylated androgens. The virilizing effects (e.g., hirsutism, acne, voice change, and alopecia) of oral androgens are typically dose and duration dependent; androgen replacement at doses < or = 10 mg once daily administered for prolonged periods (> 6 months) produces masculinization effects that generally abate with dose reduction or discontinuation of treatment. No clinical sequelae or irreversible pathophysiologic effects have been associated with any virilization that may occur. Changes in lipoprotein metabolism associated with oral estrogen-androgen use include reduced total cholesterol levels and reduced high-density lipoprotein cholesterol levels which may reduce the long-term risk of cardiovascular disease. No clinically identifiable risk with respect to other cardiovascular variables, such as blood pressure, has been associated with the longterm administration of low doses of oral androgen. With regard to liver toxicity, reports of jaundice, peliosis hepatis, and hepatocellular carcinoma are extremely rare at the dose levels of androgen used in hormone replacement therapy, although individual sensitivity to the potential hepatotoxic effects of oral alkylated and nonalkylated androgen may vary considerably. Daily dosing with oral alkylated androgen in combination with estrogen is well tolerated. Retrospective and prospective studies involving the use of androgens alone and in combination with estrogens demonstrate that concerns about the adverse effects of androgen use associated with supraphysiologic, self-escalated doses in men do not apply to the much lower doses combined with estrogens for hormone replacement in postmenopausal women. PMID- 9220206 TI - Pharmacokinetics and pharmacodynamics of midazolam given via continuous intravenous infusion in intensive care units. AB - Critically ill patients often benefit from sedation to optimize their care and their ventilatory support. Ideally, incremental doses of a drug are administered to produce the desired level of sedation without toxicity or overdose. Because metabolism and elimination of drugs are often altered in critically ill patients, knowledge of the pharmacokinetics of sedative hypnotics is essential to ensure their appropriate selection and administration. Furthermore, the administration of sedatives via continuous infusion minimizes fluctuations in drug concentrations and permits more consistent control of the patient's agitation and anxiety. Physician preference and the patient's individual requirements and underlying diseases are the primary determinants for the selection of a given sedative. Benzodiazepines are the most commonly used sedatives in critical care. Midazolam is readily distinguished from other benzodiazepines because of its rapid onset and short duration of action, low incidence of thrombophlebitis and pain on injection, and minimal cardiovascular and respiratory effects. The physiochemical properties of midazolam allow for enhanced water solubility, which limits physicochemical incompatibilities. These properties make midazolam a valuable sedative that can be given via continuous intravenous infusion in the intensive care unit. PMID- 9220207 TI - Nonsteroidal anti-inflammatory drugs, traditional opioids, and tramadol: contrasting therapies for the treatment of chronic pain. AB - The treatment of chronic pain is an important function of physicians. In the United States, available drug treatments for chronic pain currently include simple analgesics such as acetaminophen, salicylates and other nonsteroidal anti inflammatory drugs, traditional opioid drugs, and adjuvant agents (eg, antidepressants, anticonvulsants). Typically, the choice of a drug is made by balancing the indications for treatment, the clinical efficacy of the drug, and its toxicity. An understanding of the mechanism of action of these drugs helps to establish their role in therapy. Tramadol is an effective analgesic that works through a combined mechanism of weak mu receptor binding and the inhibition of serotonin and norepinephrine reuptake. Tramadol has a favorable adverse-effect profile and therefore is likely to have an important role in the management of chronic pain syndromes. PMID- 9220208 TI - Tonic-clonic seizures: a systematic review of antiepilepsy drug efficacy and safety. AB - This systematic review of studies of patients with generalized tonic-clonic seizures is an effort to evaluate whether one therapeutic agent is superior to another in terms of reducing seizures and tolerability. Recognizing that assessing relative efficacy is dependent on controlling the specific type of seizure or epilepsy treated, we restricted our review to studies in which the seizure types were clearly identified. Overall, complete control of generalized tonic-clonic seizures was achieved in 53% of treated patients. The percentage of patients who became seizure free was not significantly different with carbamazepine, phenytoin, or valproate. When patients who had a partial onset of their generalized tonic-clonic seizures were grouped, complete control was achieved in 48% with carbamazepine, 49% with phenytoin, and 52% with valproate. Overall, carbamazepine, phenytoin, and valproate appear to have similar efficacy in the treatment of tonic-clonic seizures, with complete control reported in 51%, 50%, and 55% of patients, respectively. The best response in primary generalized seizures was with valproate, with 61% reported as seizure free. Acute and dose related central nervous system side effects occurred with equal frequency with carbamazepine, phenytoin, and valproate treatment. These side effects diminished after chronic exposure. Overall, 9.9% of patients discontinued treatment due to adverse effects. The lowest incidences of clinically important side effects and rash were reported in patients treated with valproate. PMID- 9220209 TI - Valsartan, a new angiotensin II antagonist: antihypertensive effects over 24 hours. AB - This study was done to assess the antihypertensive efficacy of once-daily valsartan 20 mg, 80 mg, 160 mg, and 320 mg over 24 hours using ambulatory blood pressure monitoring (ABPM). A total of 217 adult outpatients with uncomplicated essential hypertension (office mean sitting diastolic blood pressure [DBP] of > or = 95 to < or = 115 mm Hg) participated in this multicenter, double-masked, placebo-controlled study. Patients were randomized to receive valsartan 20 mg, 80 mg, 160 mg, 320 mg, or placebo for 8 weeks. Twenty-four-hour ABPM was done at baseline and after 8 weeks of treatment. All valsartan doses produced significant decreases in average ambulatory systolic blood pressure (SBP) and DBP over 24 hours compared with placebo. A trend to greater reductions compared with placebo was observed for doses of valsartan 80 mg and greater (80 mg, -6.61 mm Hg DBP, 11.04 mm Hg SBP; 160 mg, -5.51 mm Hg DBP, -10.61 mm Hg SBP; 320 mg, -8.44 mm Hg DBP, -14.34 mm Hg SBP) compared with valsartan 20 mg (-3.52 mm Hg DBP, -5.92 mm Hg SBP). Valsartan produced consistent reductions compared with placebo during both day (> 6 AM to < or = 10 PM) and night (> 10 PM to < or = 6 AM). However, in all groups, the circadian pattern of blood pressure over 24 hours was preserved and was similar to that observed at baseline (but shifted into the normotensive range in a parallel fashion). The data show that single daily doses of valsartan 80 mg and greater provide effective control of both DBP and SBP over a 24-hour period without loss of diurnal variation. PMID- 9220210 TI - Efficacy and tolerability of doxazosin versus enalapril in the treatment of patients with mild-to-moderate hypertension. AB - The effects of 4 weeks of treatment with doxazosin or enalapril on diastolic blood pressure (DBP) and plasma lipid levels were studied in 160 patients 18 to 50 years old with mild-to-moderate hypertension. Comparing baseline measurements with measurements taken after 4 weeks, DBP was significantly reduced by 6.8 +/- 7.4 (mean +/- SD) mm Hg and 12.0 +/- 7.1 mm Hg in the doxazosin and enalapril groups, respectively. Systolic blood pressure was significantly decreased from baseline to end of treatment in both groups. There were no significant changes in heart rate from baseline to end of treatment in the doxazosin group, but there was a statistically significant decrease in heart rate in the enalapril group. High-density lipoprotein cholesterol increased statistically significantly in the doxazosin group but not in the enalapril group. A decrease in triglycerides was statistically significant with respect to the doxazosin group and was close to significance for the enalapril group. Forty-nine (62%) patients in the doxazosin group and 43 (54%) patients in the enalapril group reported at least one adverse event. Significant reductions in DBP after 4 weeks of treatment were achieved by both drugs, each taken once daily. This reduction was more pronounced in the enalapril group 24 hours postdose, with a mean final daily dose of 2.8 mg of doxazosin and 12.6 mg of enalapril. However, even relatively short-term treatment with low-dose doxazosin showed a more favorable effect on lipids than did enalapril. PMID- 9220211 TI - Itraconazole oral solution versus clotrimazole troches for the treatment of oropharyngeal candidiasis in immunocompromised patients. AB - This multicenter, open-label, third-party-masked trial compared the efficacy and safety of itraconazole oral solution (200 mg once daily) and clotrimazole troches (10 mg five times daily) in a population of immunocompromised subjects composed primarily of patients with human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS). Patients were treated for 14 days; patients who exhibited a clinical response were followed up for an additional month to document the occurrence of relapse. Efficacy was judged by changes from baseline in symptoms of oropharyngeal candidiasis (erythema, soreness/burning), extent of oral lesions, and the presence/absence of Candida species on fungal culture. A total of 162 patients were randomized, and 149 were evaluated for efficacy. The percentage of patients with negative cultures at the end of treatment was significantly greater in the itraconazole group than in the clotrimazole group (60% vs 32%, respectively). Negative culture plus clinical response was achieved in significantly more itraconazole-treated patients (53%) than clotrimazole treated patients (30%); results were similar in the subgroup of patients with HIV/AIDS. Both drugs were well tolerated, with the most frequently reported adverse events for both agents involving the gastrointestinal system. In conclusion, systemic therapy with intraconazole oral solution is efficacious and well tolerated in immunocompromised patients, including those with HIV/AIDS, when administered once daily for 14 days for the treatment of oral candidiasis. PMID- 9220212 TI - Effects of cilazapril on ventricular arrhythmia in patients with congestive heart failure. AB - Angiotensin-converting enzyme (ACE) inhibitors reduce mortality and morbidity in patients with congestive heart failure. These effects may be mediated, at least in part, by suppression of lethal ventricular tachycardia (VT). The aims of this study were to examine whether the ACE inhibitor cilazapril reduces ventricular arrhythmia in patients with congestive heart failure and, if cilazapril does reduce ventricular arrhythmia, to determine whether this reduction is associated with suppression of sympathetic nerve activity in these patients. Thirty-two congestive heart failure patients (left ventricular ejection fraction, 35 +/- 6%; New York Heart Association class II or III) with VTs (Lown grade IVa or IVb) were randomly assigned to receive either conventional therapy, consisting of diuretics and digitalis (control group), or conventional therapy plus cilazapril (cilazapril group). Twenty-four-hour ambulatory electrocardiographic monitoring was performed at baseline and after 2 months of therapy. Plasma norepinephrine levels and heart rate variability (standard deviation about the mean RR interval) were compared at baseline and after 2 months of treatment. The control group demonstrated no significant change in arrhythmia frequency after 2 months of treatment. In the cilazapril group, however, the number of ventricular couplets and VT runs was significantly decreased. In association with this reduction, plasma norepinephrine levels were decreased, and heart rate variability was increased. These results suggest that cilazapril has antiarrhythmic effects, which may be produced by suppressing high sympathetic activity, in patients with congestive heart failure. It should be noted that the study group was small and that, although ventricular dysrhythmia was reduced with therapy, it remained substantial. Further study is needed to verify these results and to determine the exact causes of the reduction. PMID- 9220213 TI - Effects of low-dose simvastatin therapy on serum lipid levels in patients with moderate hypercholesterolemia: a 12-month study. The Simvastatin Study Group. AB - The aim of this study was to investigate the safety and long-term effects on serum lipid levels of low-dose simvastatin, an inhibitor of 3-hydroxy-3 methylglutaryl coenzyme A reductase, in Japanese patients with moderate primary hypercholesterolemia. We assigned 201 patients (68 men and 133 women; mean +/- SD age, 61.3 +/- 10.2 years) with serum total cholesterol levels > or = 220 mg/dL to receive simvastatin 5 mg each evening; the treatment period was 1 year. Serum total cholesterol, triglycerides, and low-density lipoprotein (LDL) cholesterol levels decreased significantly in response to simvastatin therapy, and the changes were maintained throughout the treatment period. Mean total cholesterol decreased from 269.9 +/- 35.4 mg/dL to 215.2 +/- 34.5 mg/dL (20.3%), triglycerides decreased from 183.0 +/- 110.2 mg/dL to 155.5 +/- 88.5 mg/dL (15.0%), and LDL cholesterol decreased from 180.0 +/- 33.1 mg/dL to 130.1 +/- 35.1 mg/dL (27.7%). Total cholesterol, triglycerides, and LDL cholesterol tended to decline when the pretreatment values were higher; the critical values and the bidirectional changes of the serum lipid levels were 188.1, 109.5, and 91.6 mg/dL, respectively. Although the serum level of high-density lipoprotein cholesterol did not change significantly, it tended to increase more when the pretreatment values were lower; the "critical value" was 70 mg/dL. Nine patients experienced mild adverse events, but none discontinued simvastatin during the 12 month treatment period. We found that low-dose simvastatin therapy is effective in achieving long-term decreases in serum lipid levels and is well tolerated by patients with moderate hypercholesterolemia. Simvastatin therapy may result in normalization of serum lipid levels. PMID- 9220214 TI - Efficacy and safety of two dosing regimens of buspirone in the treatment of outpatients with persistent anxiety. AB - This randomized, double-masked, comparative study evaluated the efficacy and safety of buspirone 30 mg/d, administered twice a day (BID) or three times a day (TID), in patients with generalized anxiety disorder (GAD), commonly called persistent anxiety. Patients who participated had GAD according to criteria of the Diagnostic and Statistical Manual of Mental Disorders. Third Edition, Revised, modified to include patients for whom the symptom duration was at least 4 weeks and scored > or = 18 on the Hamilton Rating Scale for Anxiety (HAM-A). After a 7-day placebo lead-in phase, patients who continued to qualify were randomized to receive buspirone, titrated from 15 mg/d (5 mg TID) to 30 mg/d, as either a BID or TID regimen, for 8 weeks. Of the 137 patients who began the study, 120 patients were included in the data evaluation. Both buspirone BID and TID treatment groups demonstrated significant reductions in mean HAM-A total scores and improvement on Clinical Global Impression measures, with no significant differences detected between the two treatment groups for either measure at any time point. The overall incidence of adverse events was similar for both treatment groups, except for a significantly greater incidence of amblyopia in patients receiving buspirone 15 mg BID. In summary, there was no appreciable difference in efficacy or safety between buspirone 15 mg BID or 10 mg TID in patients with persistent anxiety. PMID- 9220215 TI - Bromfenac sodium, acetaminophen/oxycodone, ibuprofen, and placebo for relief of postoperative pain. AB - The objective of this double-masked, parallel-group, multicenter, inpatient study was to compare bromfenac with an acetaminophen/oxycodone combination and ibuprofen in patients who had pain due to abdominal gynecologic surgery. In the 8 hour, single-dose phase, 238 patients received single oral doses of bromfenac (50 or 100 mg), acetaminophen 650 mg/oxycodone 10 mg, ibuprofen 400 mg, or placebo. In the multiple-dose phase, 204 patients received bromfenac, acetaminophen/oxycodone, or ibuprofen for up to 5 days. In the single-dose phase, both bromfenac doses produced peak analgesic responses equivalent to acetaminophen/oxycodone, but the responses to bromfenac were longer lasting. Bromfenac produced significantly better overall (8-hour) analgesic summed scores than acetaminophen/oxycodone. Ibuprofen was less efficacious than the other analgesics. The remedication rate was lower in both bromfenac groups than in the other treatment groups. The acetaminophen/oxycodone group reported more somnolence and vomiting. Single doses of bromfenac provided analgesia at least equivalent to that of the acetaminophen/oxycodone combination, with a longer duration of action. Both doses of bromfenac and acetaminophen/oxycodone were superior to ibuprofen in this study. PMID- 9220216 TI - Ceftazidime and ciprofloxacin as empiric therapy in febrile neutropenic patients undergoing hematopoietic stem cell transplantation. AB - This pilot study was done to assess the efficacy and toxicity of intravenous ceftazidime and ciprofloxacin in patients developing febrile neutropenia while undergoing high-dose myeloablative therapy and hematopoietic stem cell transplantation (HSCT). All patients undergoing high-dose chemoradiotherapy and HSCT for leukemias, lymphomas, multiple myeloma, and solid tumors received open label ceftazidime 2 g intravenously every 8 hours and ciprofloxacin 400 mg intravenously every 12 hours if they developed fever while they were neutropenic. Success with or without modification of this regimen was defined as survival through the neutropenic period; failure was defined as death secondary to infection. Among 45 patients treated with this regimen, the success rate was 98%. Sixty-two percent (28 of 45) of the patients achieved defervescence within 48 to 72 hours and remained afebrile without regimen modification. In 16 patients (36%) the regimen was modified because of persistent fever. The combination of ceftazidime and ciprofloxacin as initial empiric antibacterial therapy in febrile neutropenic patients undergoing myeloablative therapy and HSCT appears to be highly effective and is associated with minimal toxicity. PMID- 9220217 TI - Effect of doxazosin therapy on glucose tolerance and lipid metabolism in hypertensive patients with impaired glucose tolerance. AB - The effects of long-term monotherapy with doxazosin, an alpha 1-blocker, or placebo on blood pressure (BP), glucose tolerance, and serum lipid levels were investigated prospectively in 43 hypertensive patients with impaired glucose tolerance. The levels of plasma glucose, serum lipids, fructosamine, and glycated hemoglobin A1c (Hb A1c) were determined before and during long-term (mean treatment period, 6.7 months) therapy with doxazosin (n = 23) or placebo (n = 20). A 75-g oral glucose tolerance test was performed before and during therapy. Significant decreases in both systolic and diastolic BP were maintained during doxazosin therapy; BP did not change in the placebo group. Neither fasting nor post-glucose-load venous plasma glucose levels were altered, and there was no significant change in the insulinogenic index in either group. Glucose intolerance was slightly improved with significant reductions in Hb A1c and fructosamine levels during doxazosin therapy. Serum total cholesterol (TC) and low-density lipoprotein (LDL) cholesterol levels were significantly decreased, and high-density lipoprotein cholesterol levels were significantly increased in patients treated with doxazosin. Moreover, TC, LDL cholesterol, and apolipoprotein B levels were significantly decreased in patients with hypercholesterolemia (TC > or = 5.69 mmol/L). In contrast, there were no significant changes in Hb A1c, fructosamine, and lipid levels in the placebo group. These results suggest that long-term doxazosin therapy may improve glucose and lipid metabolism in hypertensive patients. Doxazosin appears useful as an antihypertensive agent for hypertensive patients with either impaired glucose metabolism or dyslipidemia. PMID- 9220218 TI - Cost drivers in diabetes care: the problems they present and potential solutions. AB - Diabetes mellitus is a chronic disease that affects many aspects of the lives of diagnosed patients and their families, the health care industry, and society. The majority of the economic literature on diabetes addresses the cost of treating diabetes but not the outcomes of clinical interventions. The primary cost of treating diabetes is related to short-term care to achieve euglycemia and long term care associated with complication of the disease. The short-term costs of achieving euglycemia can be overshadowed by the decreased risk of long-term complications. The difficulty of providing care for a chronic disease such as diabetes arises from the high short-term costs of clinical interventions, the positive benefits of which may not be realized for many years. The results of the Diabetes Control and Complications Trial show a correlation between the intensive treatment of diabetes and a decreased risk of the development of long-term complications. Whether intensive treatment is practical, effective, and cost effective in a real-world setting is a topic for further study. In the meantime, health care providers with a good knowledge of the clinical and economic elements of available therapeutic options can develop individualized care regimens for their patients with diabetes that are high quality and cost-effective. PMID- 9220219 TI - Pharmacoeconomic applications of meta-analysis for single groups using antifungal onychomycosis lacquers as an example. AB - This paper presents a method for summarizing clinical success rates across studies or arms of studies. Topical lacquers for the treatment of onychomycosis are used to illustrate the approach. A modification of Cochran's method, later modified by Der-Simonian and Laird, is presented in a stepwise fashion. A summary point estimate weighted by both within- and between-study variance is produced, along with a 95% confidence interval. This method can be applied to any set of proportions. In the example, the success rate of ciclopirox lacquer was 81.6% (standard error, 3.5%) and of amorolfine lacquer was 71.4% (standard error, 2.3%). A stricter definition of success (ie, cure only) produced success rates of 31.3% (standard error, 2.4%) and 35.8% (standard error, 1.9%), respectively. This stepwise approach for combining single proportions is suggested as a useful method for summarizing success rates across different trials. PMID- 9220220 TI - Outcomes and costs of positron emission tomography: comparison of intravenous adenosine and intravenous dipyridamole. AB - The objective of this study was to compare the cost of intravenous adenosine and intravenous dipyridamole in positron emission tomography (PET) in patients with coronary artery disease. A retrospective, open-label, case-control, cost effectiveness analysis was performed in the out-patient nuclear medicine department of a university hospital. Thirty-six patients underwent dipyridamole PET, and 72 matched patients underwent adenosine PET. A cost-effectiveness analysis was conducted using a direct cost accounting approach to estimate institutional costs. Key costs evaluated included acquisition cost, administration cost, monitoring cost, cost of management of side effects, and cost of follow-up care. The total cost of adenosine PET and dipyridamole PET was divided by their respective predictive accuracies to provide a total cost adjusted for efficacy. Adenosine increased heart rate and lowered systolic blood pressure to a significantly greater extent than dipyridamole. The number of patients experiencing adverse drug reactions was significantly greater for adenosine (82%) than for dipyridamole (67%), but the frequency of prolonged (> 5 minutes) and late-onset side effects was significantly greater for dipyridamole than for adenosine. The frequency of side effects requiring medical intervention was also significantly greater for dipyridamole (53%) than for adenosine (6%). Although adenosine had a significantly greater acquisition cost than dipyridamole, costs of monitoring, management of side effects, and follow-up care were significantly less for adenosine than for dipyridamole. As a result, the total cost of using dipyridamole is significantly greater ($928.00 per patient) than the total cost of using adenosine ($672.00 per patient). Based on these results, adenosine may be the drug of choice for pharmacologic vasodilation for PET. PMID- 9220221 TI - Cost-effectiveness of initial therapy with 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors to treat hypercholesterolemia in a primary care setting of a managed-care organization. AB - From January 1994 through May 1995, Prudential HealthCare-North Texas prospectively studied 299 member patients diagnosed with hypercholesterolemia for whom pharmacotherapy with one of four 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, also known as statins, was prescribed. The purpose of this study was to measure the relative cost-effectiveness (CE) of these drugs in a real-world setting. This study provides information to assist decision makers in managed-care organizations (MCO) in making formulary selections. The study used a prospective, randomized, balanced cohort design, examining patients who had been prescribed initial therapy with a statin drug as monotherapy. Costs (direct medical and indirect costs) and effectiveness (percent reduction in low-density lipoprotein cholesterol levels) were based on approximately the first 6 months of initial therapy. Both the MCO and patient perspectives were considered. In the base case, mean CE ratios were significantly lower for fluvastatin compared with lovastatin, pravastatin, and simvastatin from both the managed-care perspective and the patient perspective. Sensitivity analysis did not alter the CE conclusions, even under conditions of varying cost structures. Although differences were found in the effectiveness of lovastatin, pravastatin, and simvastatin measured in this study versus efficacy measured for these drugs in controlled clinical trials, sensitivity analysis suggests that these differences alone do not determine the superior CE of fluvastatin. Finally, this study supports the idea that well-designed formularies should consider drug CE (based on safety, effectiveness, and cost) and that integration of the pharmacy benefit management with other medical management is essential. These results provide evidence that fluvastatin may represent a more cost-effective formulary choice among statin products used for initial monotherapy of hypercholesterolemia. PMID- 9220222 TI - Estrogen and progestin components of oral contraceptives: relationship to vascular disease. AB - Recently, new information has been published about: a) the relationship between combination oral contraceptives (OCs), estrogen dose, cigarette smoking, and the risk of myocardial infarction (MI) and stroke; and b) the effect of different progestins on the risk of venous thromboembolism (VTE). We review the epidemiologic data. Regardless of age, in the absence of smoking, use of sub-50 micrograms OCs is not associated with any meaningful increase in risk of MI or stroke. If the small, statistically nonsignificant elevations in risk for these diseases are assumed (for the sake of argument) to be causal, then the incidence of MI and stroke associated with use of OCs containing less than 50 micrograms ethinyl estradiol (EE) would be approximately 2 per 100,000 per year. For women less than 35 years of age who do not smoke or do not have a history of hypertension, the risk would be even lower. Any woman over the age of 35 who smokes should be advised to use a non-estrogen or nonhormonal contraceptive. There are now two reports, from jick et al. and Lewis et al., that demonstrate that the relative risk of MI is certainly no greater for users of OCs containing desogestrel or gestodene than for users of OCs containing older progestins. In fact, both show reduced relative risks for the newer progestins compared to the older ones. With respect to progestins, four recent epidemiologic studies have indicated a twofold increased risk of nonfatal VTE with use of OCs containing desogestrel or gestodene compared with levonorgestrel. A fifth report, which showed an increased relative risk for norgestimate, is based on use among only 19 cases and 31 controls and is not statistically significant. As the authors themselves caution and as subsequent follow-up analyses and editorials conclude, these studies do not provide evidence for a cause-and-effect relationship between OCs containing desogestrel or gestodene, and VTE. The recommendation with respect to desogestrel- and gestodene-containing OCs is that no change in prescribing practices is warranted for either current or new-start patients. There is a growing body of evidence demonstrating that OCs containing 30 or 35 micrograms of EE have lower risks of MI, stroke, and VTE than higher dose OCs. However, there is no epidemiologic study that demonstrates a greater risk of vascular events among women using OCs containing 30 or 35 micrograms EE compared with preparations containing 20 micrograms EE. Users of sub-50 micrograms OCs of any age have no clinically meaningful increase in incidence of MI or stroke compared with non-OC users. This is also true for smokers under the age of 35 years who use OCs. However, smokers over the age of 35 years who use OCs still have an unacceptably high incidence rate of MI and stroke and should not use combination OCs. Sub-50 micrograms OCs of all types are associated with a small excess risk of VTE, about 15 per 100,000 events per year. Until there is biologic explanation of the twofold greater risk of VTE in users of OCs containing desogestrel or gestodene compared with users of those containing older progestins, this association should not be accepted as one of cause and effect. PMID- 9220223 TI - Pharmacokinetics of the new progestogens and influence of gestodene and desogestrel on ethinylestradiol metabolism. AB - The purpose of the present report is to summarize the most important pharmacokinetic features of the new progestogens. In addition, the question of whether or not gestodene, in comparison to desogestrel, has an influence on the pharmacokinetics of ethinylestradiol (EE2) will be addressed. PMID- 9220224 TI - Outpatient laparoscopic interval female sterilization. AB - A 23-year retrospective review of laparoscopic sterilization in Ramathibodi Hospital, Bangkok, Thailand, is reported. A total of 9041 cases of outpatient laparoscopic interval female sterilizations were done from January 1973 to December 1995. Intraoperative complications occurred in 35 cases (0.39%) and hospital admissions totalled 65 cases (0.72%). Adnexal injuries were the most frequent complication. There was one case of death from anesthetic complication. Management and prevention of complications are discussed. PMID- 9220225 TI - Determinants for contraceptive use in young, single, Danish women from the general population. AB - Determinants for contraceptive use were studied in 5031, non-pregnant women aged 20-29 years from the general population in Denmark. Most women (72%) had never been pregnant, 34% had a history of a sexually transmitted disease, and 22% had ever had a legal abortion. Current contraception was most frequently condoms (60%) or oral contraceptives (33%). Among the women who used OCs or IUD, 32% reported additional condom use (double contraception). Important predictors of using one contraceptive method were lifetime number of sexual partners, parity, and age at first sexual intercourse for condoms and age for oral contraceptives. Also, women with a previous legal abortion were more likely to use condoms currently and women with a history of STDs were less likely to use condoms, but more likely to use OCs. Lifetime number of sexual partners was the only predictor of double contraception. Our data suggest a potential for reducing the number of unintended pregnancies and STDs in single women by increasing the information about the double principle in contraception. PMID- 9220226 TI - Comparative clinical evaluation of the effect on carbohydrate and lipid metabolism of two norethisterone-containing hormonal contraceptives: Mesigyna and TriNovum. AB - The effect on carbohydrate and lipid metabolism of two hormonal contraceptive preparations containing norethisterone (commercially known as Mesigyna and TriNovum) was studied in a total of 60 women, before and after 6 months of treatment. Carbohydrate metabolism was evaluated by means of a euglycemic glucose clamp test; lipid metabolism was monitored by measuring total cholesterol, HDL cholesterol, LDL-cholesterol, VLDL-cholesterol, and triglycerides. The two groups were properly matched with the exception of pretreatment levels of cholesterol and LDL-cholesterol. At the end of treatment, no difference was found within or between groups in fasting glucose and insulin levels and in glucose rate of disappearance. A significant increase in total cholesterol, HDL-cholesterol, and VLDL-cholesterol was found in both groups at the end of the treatment period; in addition, TriNovum caused a significant increase also in triglycerides. In conclusion, the safety of both preparations with regard to carbohydrate metabolism was confirmed using the most accurate method available; furthermore, changes in lipid metabolism were such as to have little clinical significance. PMID- 9220227 TI - Birth control: some experiences from Denmark. AB - This article discusses that an effective birth control exists in Denmark today. Birth control is considered as a means for the couple not only to reach the wished-for family size by limiting their number of children but also to decide when to have children. Since 1973, women in Denmark can have an induced abortion on request before the end of the 12th week of pregnancy. An increasing proportion of women applying for induced abortion have no children, indicating that induced abortion is a means to obtain an effective postponement of first birth. The paper concludes that birth control is primarily accomplished by use of contraceptives and as an effect of widespread use, teenagers have managed to diminish their total rate of pregnancy rather dramatically. PMID- 9220228 TI - Return of fertility after discontinuation of the once-a-month injectable contraceptive Cyclofem. AB - The objective of this study was to evaluate the return of fertility in women who used Cyclofem as a contraceptive method during the introductory studies conducted in Brazil, Chile, Colombia, and Peru. From these four cohorts, 101 women were eligible for the study. Thirty-one were not included in the study either because they refused to be interviewed, had initiated another contraceptive method the month after discontinuation, or were unable to be contacted. A total of 70 women were included in the study. Our results showed that the return to fertility rate after the discontinuation of Cyclofem was 1.4 per 100 women at the end of the first month and reached 82.9 at one year. More than 50% were pregnant at 6 months. Fifty-one (94.4%) pregnancies ended in a live birth, two were spontaneous first trimester abortions, and one was a hydatidiform mole. Return of fertility was not related to the woman's age at the time of discontinuation, her weight, or the number of Cyclofem injections. In conclusion, fertility is restored by 1 month following Cyclofem discontinuation. Users and potential users should be counseled regarding the rapid return of fertility after discontinuing this method of contraception. PMID- 9220229 TI - Effects of oral buserelin on urinary LH secretion in male infants. AB - In recent years, several potent gonadotropin-releasing hormone (GnRH) analogues have become available for female contraception and one of them (buserelin) has been tested in lactating women. However, the possible effects on infants due to the transference of the analogue through breast milk have not been studied. The present work evaluated the effect of oral buserelin on urinary LH secretion in male infants. A total of 19 healthy full-term boys (aged 2-4 months) were included in the study. Infants received orally a single dose of a GnRH agonist mixed with breast milk. Urine samples were collected prior to, and 4-6 and 24 h after treatment for LH measurement. The results disclosed a significant increase in LH urine level in the sample taken 4-6 h after buserelin administration. Twenty-four hours after GnRH agonist ingestion, the LH level returned to baseline level. The present study demonstrated that GnRH analogue administered orally to infants escapes from gastrointestinal inactivation and induces a significant rise in LH levels 4-6 h after treatment. PMID- 9220230 TI - Nitrites and L-citrulline levels in copper intrauterine device users. AB - The mechanism of copper in limiting intrauterine infections in intrauterine device (Cu IUD) users is poorly understood. Copper ions may enhance the release of reactive oxygen radicals, which in turn decrease the release of reactive nitrogen intermediates (RNI). RNI are known to have bactericidal effect. The present study compares the levels of RNI prior to Cu-T insertion and at different post-insertion intervals up to 12 weeks. The decrease in RNI was evident by one week and continued until 12 weeks. Therefore, the bactericidal effect of copper in IUD is via reactive oxygen intermediates. The superoxide anion inactivates this active intermediate nitric oxide. Therefore, excess of superoxide radical will markedly shorten the half-life of nitric oxide but will not prevent its conversion to nitrites and nitrates. PMID- 9220231 TI - Post-Norplant implants insertion anaphylactoid reaction: a case report. PMID- 9220232 TI - The role of cultures in the management of ulcerative keratitis. PMID- 9220233 TI - The role of cultures in the management of ulcerative keratitis. AB - PURPOSE: To ascertain the importance of routine cultures and gram stains in the management of ulcerative keratitis. METHODS: We retrospectively reviewed 119 consecutive corneal ulcers seen at Sinai Hospital of Baltimore. Cultures were obtained of the corneal ulcer and of the lids and conjunctivae of both eyes. Gram stains were performed by the hospital microbiology department on corneal scrapings from each ulcer. RESULTS: Positive corneal cultures were obtained from 56 eyes (47.1%). Initial antibiotic therapy was changed based on culture results in 14.3% of culture-positive eyes that demonstrated a worsening clinical course. Gram stains were negative in all cases. The sensitivity and specificity of the lid and conjunctival cultures were determined. CONCLUSIONS: Corneal cultures are important in the management of ulcerative keratitis. Lid and conjunctival cultures have low sensitivity and specificity. PMID- 9220234 TI - Long-term graft survival in patients with flexible open-loop anterior-chamber intraocular lenses. AB - PURPOSE: To evaluate the long-term graft survival and complications of flexible, open-loop anterior-chamber intraocular lenses in patients with penetrating keratoplasty for pseudophakic or aphakic bullous keratopathy. METHODS: We reviewed charts of all consecutive patients who underwent penetrating keratoplasty for pseudophakic or aphakic bullous keratopathy combined with implantation of a flexible, open-loop, anterior-chamber intraocular lens at our institution between 1983 and 1988. One-hundred one eyes of 99 patients were evaluated. Graft-survival rates were calculated by using the Kaplan-Meier actuarial method. RESULTS: Mean follow-up was 49.8 months (range, 1-144). The probability of graft survival at 1, 2, 4, 6, and 8 years was 93, 87, 78, 65, and 65%, respectively. A total of 25 (24.8%) grafts failed. Progressive corneal edema without signs of rejection was the most common finding in patients with failed grafts (10 eyes, 40%). The most frequent complication observed was newly diagnosed or worsening of preexisting glaucoma (46 eyes, 45.5%). CONCLUSIONS: Our long-term results support flexible, open-loop anterior-chamber intraocular lenses as a reasonable option, at the time of penetrating keratoplasty, in patients with pseudophakic and aphakic bullous keratopathy. PMID- 9220235 TI - Ganciclovir ophthalmic gel (Virgan; 0.15%) in the treatment of herpes simplex keratitis. AB - PURPOSE: Ganciclovir is a broad-spectrum virustatic agent. Its efficacy and safety after ocular application have been demonstrated in studies of herpetic keratitis in rabbits. Two strengths of ganciclovir gel (0.05 and 0.15%) were compared with 3% acyclovir ointment in the treatment of superficial herpes simplex keratitis in humans. METHODS: Two multicenter randomized clinical trials were carried out in Africa (Trial 1) and Europe (Trial 2). Sixty-seven patients (Trial 1) and 37 patients (Trial 2) from herpetic ulceration were recruited. RESULTS: The results showed no statistically significant difference between the treatment groups, although the healing rates tended to be better in the group receiving 0.15% ganciclovir gel, with healing rates of 85% (Trial 1) and 83% (Trial 2) as compared with 72% (Trial 1) and 71% (Trial 2) in the group receiving acyclovir ointment. Local tolerance was found to be superior with the gel formulation of ganciclovir with fewer complaints of discomfort (stinging, burning) or blurred vision after application of the drug. Systemic absorption of the drug was low. No hematologic changes were detected. CONCLUSIONS: These findings support the efficacy of ganciclovir gel in the treatment of ulcerative herpes simplex keratitis and demonstrate its superior local tolerance when compared with acyclovir ointment. PMID- 9220236 TI - Ocular flora of patients with AIDS compared with those of HIV-negative patients. AB - PURPOSE: To determine whether there are quantitative or qualitative differences in the ocular flora of patients with acquired immunodeficiency syndrome (AIDS) compared with human immunodeficiency virus (HIV)-negative patients. METHODS: Forty patients with AIDS and 42 HIV-negative controls were sex and age matched. All subjects had a detailed anterior segment examination, including Schirmer's test, rose bengal staining, and quantitative cultures of the conjunctiva and lids. Statistical evaluation of the relation between AIDS, keratoconjunctivitis sicca (KCS), and ocular flora was performed. RESULTS: No differences were observed in the types or numbers of organisms isolated from the conjunctiva or lids of patients with AIDS and HIV-negative subjects. Ocular flora was not influenced by use of systemic antibiotics, level of immunosuppression as measured by CD4 lymphocyte counts, KCS, or other ocular-surface disease. One AIDS patient was colonized by large numbers of Haemophilus influenzae OU with minimal clinical signs of inflammation or infection. CONCLUSION: There do not appear to be any differences in the ocular flora of HIV-negative patients and patients with AIDS. Presence of KCS and level of immunosuppression do not appear to affect the ocular flora in patients with AIDS. PMID- 9220237 TI - Evaluation of diclofenac sodium 0.1% ophthalmic solution in the treatment of ocular symptoms after bilateral radial keratotomy. AB - PURPOSE: Patients frequently have ocular pain, photophobia, foreign-body sensation, and burning/stinging after radial keratotomy. This study was a prospective, randomized, double-masked, multicenter, fellow-eye comparison of diclofenac sodium (Voltaren Ophthalmic, 0.1% solution) and placebo for controlling these ocular symptoms after bilateral radial keratotomy. METHODS: Patients who were pain free in both eyes before surgery were randomly assigned to treatment with diclofenac sodium in one eye and placebo in the other. One drop of each masked trial medication was administered 30-60 min before surgery, 5 min and 6 h after surgery, at bedtime on the day of surgery, and four times daily for 2 additional days. Patients evaluated ocular symptoms in each eye 0.5, 1, 2, 4, 6, 24, and 48 h after surgery and provided a global evaluation 6, 24, and 48 h after surgery. For each assessment, the difference in scores between eyes was analyzed by using a paired t test. RESULTS: Diclofenac sodium was significantly (p < 0.001) superior to placebo in controlling each ocular symptom at each interval after surgery and for patient global assessments 6, 24, and 48 h after surgery. CONCLUSION: Diclofenac sodium 0.1% ophthalmic solution is clinically effective in controlling adverse ocular symptoms occurring after bilateral radial keratotomy. PMID- 9220238 TI - Indications for penetrating keratoplasty in Canada, 1986-1995. AB - PURPOSE: To examine the leading indications for penetrating keratoplasty. METHODS: We retrospectively performed a chart review of 904 cases of penetrating keratoplasty over a 10-year period (1986-1995). Where possible, the clinical indication was corroborated by the pathological report. RESULTS: Leading indications included pseudophakic bullous keratopathy (PBK; 28.5%), regraft (22.4%), keratoconus (10.0%), Fuchs' dystrophy (7.6%), aphakic bullous keratopathy (ABK; 6.1%), herpetic keratopathy (4.2%), bacterial infection (4.2%), physical trauma (4%), interstitial keratitis (3.7%), and corneal scarring (2.8%). The most common type of intraocular lens found in patients with PBK was the anterior-chamber lens [AC IOL, 71.6%; posterior-chamber lens (PC IOL), 16%; iris plane IOL (IP IOL; 12.5%]. In our series, we noted decreasing trends in the incidences of ABK (p < 0.001) and PBK (p < 0.04) and an increasing trend in the incidence of regraft (p < 0.0083). Our data do not show statistically significant changes in the incidences of Fuchs' dystrophy and keratoconus. Gender differences were present for Fuchs' dystrophy with female exceeding male patients by a 3:1 ratio (p < 0.001). Male exceeded female patients by 2:1 for keratoconus (p < 0.001). Male exceeded female patients by 3:1 for trauma (p < 0.001). CONCLUSIONS: In this series, the leading indications for penetrating keratoplasty were found to be generally in agreement with the data reported in recent literature. Variations may reflect the wide variations in practice and referral patterns. PMID- 9220239 TI - Deposits of topical norfloxacin in the treatment of bacterial keratitis. AB - PURPOSE: Fluoroquinolone solutions are widely used in therapy of bacterial keratitis. These drugs are safe, and their ocular side effects are mild and not serious in nature. The most frequent untoward effect associated with ciprofloxacin therapy has been a white crystalline deposit. This deposit has not been reported after using the other commercially available quinolones, ofloxacin and norfloxacin. METHODS: We present three cases of norfloxacin deposits after treatment of bacterial keratitis. RESULTS: In the three cases, the norfloxacin was substituted for another antibiotic effective against the bacteria, and the precipitate spontaneously resolved in all cases within a few days with no untoward ocular effect. CONCLUSION: Crystalline corneal deposits can develop during topical norfloxacin therapy. The exact factors contributing to the formation are unknown, although the differences in the tear pH/solubility could be involved. Clinicians should be aware of this ocular side effect. PMID- 9220241 TI - The effects of age, gender, and fluid dynamics on the concentration of tear film epidermal growth factor. AB - PURPOSE: To identify the relationship between epidermal growth factor (EGF) concentration in human tears and clinical tear-flow parameters and how these vary with age and gender. METHODS: Tear samples were collected with minimal stimulation from 68 healthy and asymptomatic adults (33 men, 35 women), aged 21 88 years. EGF concentrations were determined by sandwich enzyme-linked immunosorbent assay (ELISA) in 65 cases. Schirmer tests were performed without anesthesia, and the clearance of fluorescein from the tear film assessed. The Tear Function Index (TFI) was calculated from these values. RESULTS: There were approximately equal numbers of male and female subjects with a similar age distribution for each gender (48 +/- 3 and 51 +/- 3 years, mean +/- SEM, respectively). Ninety percent of tear EGF concentrations were between 0.75 and 7.1 ng/ml. Tear EGF level correlated significantly with Schirmer I value, but not with age. Schirmer I value correlated with tear clearance [LN(TCR)] but not with age. Tear EGF concentrations were significantly higher for men (3.4 +/- 0.3 ng/ml) than for women (2.4 +/- 0.3 ng/ml; p = 0.043). CONCLUSIONS: EGF concentrations is tear samples from normal humans were found to correlate with gender and Schirmer I value but not with tear clearance. PMID- 9220240 TI - Topical anesthetic abuse ring keratitis: report of four cases. AB - PURPOSE: We present the clinicopathologic correlations of two case and two other clinical cases of topical anesthetic abuse keratopathy that were originally diagnosed as Acanthamoeba keratitis because of ring keratitis presentation and characteristic history. METHODS: Four patients who were referred to us with suspected Acanthamoeba keratitis are included. Each was initially treated for amoebic keratitis, by using established protocols, and only later was the true origin (topical anesthetic abuse) uncovered. The clinical and surgical histories, pathologic analysis of the corneal specimens, and follow-up of < or = 4 years are included. RESULTS: Our four cases show another cause for ring infiltration of the cornea. Two cases resulted in corneal transplantation and multiple other medical or surgical treatments in an attempt to restore vision but had poor outcomes of finger-counting vision. Two other cases responded to intensive medical treatments with return of useful vision. Evaluation of the surgical specimens revealed a previously unpublished finding of near total cell death within the corneal stroma. CONCLUSION: Topical anesthetic abuse resulting in sight-threatening keratitis may be seen as a masquerade syndrome in many cases. Because of the often poor outcome, we must be aware of this entity, prevent abuse, and be vigilant in our prohibition of topical anesthetic for any therapeutic use. PMID- 9220242 TI - Extrusion of abnormal endothelium into the posterior corneal stroma in a patient with posterior polymorphous dystrophy. AB - PURPOSE: To report the presence of abnormal endothelium that extruded into the posterior corneal stroma in a patient with posterior polymorphous dystrophy. METHODS: The corneal button of a man who underwent penetrating keratoplasty for posterior polymorphous dystrophy was examined by light and electron microscopy. Immunoperoxidase staining for cytokeratins, vimentin, and the endothelial antigen recognized by monoclonal antibody 2B4.14.1 antigen was performed. Two-color immunofluorescence staining for simultaneous detection of cytokeratins and 2B4.14.1 antigen was also done. RESULTS: Much of the endothelium had characteristic features of epithelium-like cells, and abnormalities in Descemet's membrane were present. Curious oval and slit-like spaces in the posterior stroma were lined by epithelium-like endothelial cells and were continuous with the anterior chamber through defects in Descemet's membrane. CONCLUSION: These abnormalities in the posterior stroma have not previously been described in histopathologic reports of posterior polymorphous corneal dystrophy and are likely an unusual variation in the spectrum of this hereditary disorder. PMID- 9220243 TI - Polyhexamethylene biguanide (PHMB) in the treatment of experimental Fusarium keratomycosis. AB - PURPOSE: We wanted to determine whether topical polyhexamethylene biguanide (PHMB) 0.02% was effective in the treatment of experimental Fusarium keratomycosis in rabbits. METHODS: Fusarium solani keratomycosis was induced in the eyes of 12 New Zealand white rabbits. The rabbits were treated with PHMB 0.02% in one eye and placebo in the other eye for 6 days. The rabbits were evaluated in a masked fashion using a standardized system for clinical progression of the disease. Then the corneas were trephined and growth of F. solani in colony-forming units per milliliter (CFU/ml) determined. RESULTS: Clinical evaluation demonstrated no significant mean difference (p > 0.10) in clinical scores between treated and control eyes on day 6 (0.583 +/- 2.503). There was a significant mean CFU difference (p = 0.06) between treated eyes and control eyes (182.5 +/- 314.44). Seven of 12 eyes (58%) in the PHMB group exhibited no growth, whereas two of 12 (17%) eyes reported no growth in the control group. One of 12 eyes (8%) reported > 100 CFU in the PHMB group, whereas seven of 12 eyes (58%) reported > 100 CFU in the control group. CONCLUSIONS: PHMB 0.02% was effective in significantly reducing the fungal growth in our rabbit model of Fusarium keratomycosis. The future role of PHMB in the treatment of Fusarium keratitis needs to be further evaluated. PMID- 9220244 TI - A novel method of tear collection: comparison of glass capillary micropipettes with porous polyester rods. AB - PURPOSE: To develop a rapid, user-friendly method of tear collection to facilitate tear-protein analysis. METHODS: Tears were collected from a total of 19 normal volunteers without evidence of ocular-surface disease with either porous polyester rods or glass-capillary micropipettes. Tear-collection rate and recovery of two tear proteins, epidermal growth factor (EGF, low abundance) and lactoferrin (LFR, high abundance) were compared between polyester rods and glass capillary micropipettes. The recovery of LFR and EGF and the stability of these proteins after storage at -70 degrees C were quantitated by specific monoclonal enzyme-linked immunosorbent assay (ELISA). RESULTS: Polyester rods collected tears an average of 3.9-fold faster than glass-capillary micropipettes (p < 0.001). Both methods were comparable in efficacy of protein recovery. The polyester rods demonstrated a trend toward enhanced recovery, but this difference was not statistically significant (p = 0.12, LFR; p = 0.055, EGF). Analysis of the reliability and reproducibility of the tear-collection assay system revealed that ELISA analysis is highly reproducible, but there is significant day-to-day variation in tear-protein levels of both LFR and EGF for a given volunteer. Both LFR and EGF displayed a trend toward enhanced detection by ELISA shortly after freezing at -70 degrees C and slow decay after storage at -70 degrees C for up to 72 and 105 days, respectively. After stimulation of reflex tearing via the nasolacrimal reflex, LFR levels remained relatively constant, whereas EGF levels for most patients declined and then plateaued. CONCLUSIONS: Polyester rods provide a more rapid, user-friendly alternative to glass-capillary micropipettes for the collection and analysis of tear fluid and tear proteins. Polyester rods may have greater clinical utility, facilitating routine analysis of the preocular tear film. PMID- 9220245 TI - Gene transfer to ex vivo stored corneas. AB - PURPOSE: We examined the efficiency and kinetics of recombinant adenovirus vector mediated gene transfer to rat and rabbit cornea in culture ex vivo. METHODS: A recombinant replication-defective adenovirus was used to transfer a lacZ marker gene to whole rat and rabbit corneas in culture. Histochemistry was used to localise transgene expression and a colorimetric assay to quantify recombinant protein expression. RESULTS: After infection with recombinant virus and culture for 3 days, high-efficiency gene transfer was found, with expression in most endothelial cells of both species. Minimal expression was found in other corneal cell types. On histochemistry, longer duration of expression was found in rat than in rabbit endothelium. In both rat and rabbit cornea, highest levels of recombinant protein were found at days 3-7 after incubation with virus, decreasing to low or undetectable levels at 21 days. CONCLUSION: Adenovirus vectors allow high-efficiency transgene expression in cornea, largely restricted to the endothelial cells of ex vivo cultured cornea. Kinetics of expression differ according to the species of cornea studied, a factor that must be considered if this vector is used in further studies. PMID- 9220246 TI - IL-1 upregulates keratinocyte growth factor and hepatocyte growth factor mRNA and protein production by cultured stromal fibroblast cells: interleukin-1 beta expression in the cornea. AB - PURPOSE: To determine whether interleukin 1 beta (IL-1 beta) messenger RNA (mRNA) and protein were expressed in corneal cells and to examine the effects of IL-1 alpha and IL-1 beta on the expression of hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF) mRNAs and proteins in corneal stromal fibroblasts. METHODS: IL-1 beta mRNA was detected by reverse transcriptase polymerase chain reaction (RT-PCR). IL-1 beta protein was detected by immunohistologic tests. Changes in the expression of HGF and KGF mRNAs and proteins in response to stimulation of cultured corneal stromal fibroblasts with IL-1 alpha and IL-1 beta were monitored by Northern and Western blotting, respectively. RESULTS: IL-1 beta mRNA is expressed in human primary cultured corneal epithelial, stromal fibroblast, and endothelial cells. IL-1 beta protein was detected in epithelium and endothelium in fresh frozen human and rabbit corneal tissue. Little, if any, IL-1 beta was detected in the unwounded corneal stroma. IL-1 alpha and IL-1 beta at 10 ng/ml upregulated the levels of HGF and KGF mRNAs and proteins in cultured human corneal fibroblasts. CONCLUSIONS: IL-1 alpha and IL-1 beta may serve as key modulators in an epithelial-stromal regulatory loop in the cornea. These data and previously published observations support the hypothesis that corneal epithelial wounding releases IL-1 alpha and IL-1 beta from epithelial cells; these cytokines in turn upregulate HGF and KGF mRNA and protein levels in keratocytes, and HGF and KGF released by the keratocytes modulate healing of the wounded corneal epithelial cells by regulating proliferation, motility, and differentiation. PMID- 9220247 TI - A new classification system predicting keratomalacia after trauma in vitamin A deficiency: sodium citrate does not prevent disease progression. AB - PURPOSE: The purpose of this study was to develop a classification system to predict keratomalacia after trauma in vitamin A-deficient eyes and to determine whether citrate impedes polymorphonuclear leukocyte infiltration into the cornea, thus preventing keratomalacia. METHODS: Preliminary classification studies showed that a 7.0-mm corneal epithelial scrape, before clinical findings of corneal xerosis, did not induce keratomalacia. Primary studies were conducted concurrently on the same animals to develop the classification system and test the effect of citrate in vitamin A deficiency. A 7.0-mm corneal epithelial scrape was performed on vitamin A-deficient eyes in various stages of corneal xerosis and treated as follows. Experiment 1: group 1, 10% citrate drops; group 2, phosphate buffer solution (PBS) drops; experiment II: group 3, drops and subconjunctival injection of 10% citrate; group 4, drops and subconjunctival injection of PBS. RESULTS: Corneal abrasion in eyes with 2+ corneal xerosis yielded keratomalacia in 50% of cases; the remainder healed with xerotic epithelium. Eighty-three percent of eyes with > 2+ xerosis developed keratomalacia after corneal abrasion, whereas only 7.1% of eyes with < 2+ xerosis advanced to this stage. In experiment I, 27% of citrate-treated eyes and 38% of PBS-treated eyes developed keratomalacia (not significant). In experiment II, two of six citrate-treated eyes perforated and one eye developed keratomalacia. One of six control PBS eyes perforated and four developed keratomalacia. CONCLUSION: We correlated the degree of corneal xerosis with the occurrence of keratomalacia after corneal trauma. This led to the development of a classification scale that is of research and clinical significance. Additionally, citrate did not significantly reduce keratomalacia or perforation in the vitamin. A-deficient eye. PMID- 9220248 TI - Bilateral peripheral ulcerative keratitis associated with pyoderma gangrenosum. AB - PURPOSE: A 37-year-old Hispanic man with a history of chronic myelogenous leukemia was first seen with fever and preseptal cellulitis of this right orbit after a tooth extraction. METHODS: The patient subsequently developed bilateral, severe peripheral ulcerative keratitis. He was treated with systemic antibiotics for a presumed underlying infectious cause. Several painful, necrotizing skin lesions developed over his face, trunk, and extremities. RESULTS: Repeated skin biopsies of the necrotizing lesions were consistent with pyoderma gangrenosum. Both the skin lesions and peripheral ulcerative keratitis responded dramatically to systemic prednisone. CONCLUSIONS: Pyoderma gangrenosum should be included in the differential diagnosis of peripheral ulcerative keratitis. PMID- 9220249 TI - Pellucid marginal degeneration with superior corneal thinning. AB - PURPOSE: Pellucid marginal degeneration is a noninflammatory thinning disorder typically involving the inferior cornea. We describe a patient with superior corneal thinning similar to classic pellucid marginal degeneration. METHODS: An 80-year-old man was evaluated for high astigmatism. RESULTS: The right superior cornea had a prominent band of thinning with ectasia. Both inferior corneas had characteristic zones of thinning without inflammation. CONCLUSIONS: Superior pellucid marginal corneal degeneration should be considered in the differential diagnosis of superior corneal ectatic disorders. PMID- 9220250 TI - Progressive nonulcerative paracentral keratolysis associated with elevated corneal metalloproteinases. AB - PURPOSE: We report a patient with progressive idiopathic, nonulcerative, noninflammatory, avascular, bilateral, paracentral and peripheral corneal thinning monitored for 13 years. METHODS: Because of progressive corneal thinning, the patient underwent several surgical procedures, including an arcuate lamellar keratectomy with suturing, bilateral 15-mm diameter onlay lamellar corneoscleral epikeratoplasties, and removal of interface epithelial tissue. Over time, the keratolysis also thinned the donor stroma, requiring a lamellar tectonic graft. A biopsy was performed of the patient's cornea and conjunctiva, and the tissue was analyzed for proteolytic enzymes. RESULTS: Increased quantities of matrix metalloproteinases (57 and 63 kDa) were extracted from the patient's normal-appearing and abnormal corneal samples but not from adjacent conjuctiva and sclera or normal controls. This is the first reported case with these clinical and laboratory findings. CONCLUSION: A previously undescribed progressive idiopathic paracentral keratolysis is associated with increased quantities of matrix metalloproteinases. Clinical management requires tectonic corneal surgery. PMID- 9220251 TI - Brown-McLean syndrome occurring in a corneal graft. AB - PURPOSE: To present a case of Brown-McLean syndrome occurring in a transplanted cornea. METHODS: Retrospective case report. RESULTS: A patient had penetrating keratoplasty performed for bullous keratopathy 6 years after intracapsular cataract extraction in the right eye. She developed atypical Brown-McLean syndrome in the left eye 6 years after extracapsular cataract extraction with pupil-support intraocular lens (IOL) insertion and later IOL removal. The clear corneal graft in the right eye developed peripheral stromal and epithelial edema with pigmentation of the endothelium, consistent with Brown-McLean syndrome 9 years after keratoplasty. CONCLUSION: Brown-McLean syndrome can occur in a grafted cornea, although the features may differ from those seen in ungrafted corneas. PMID- 9220252 TI - Self-induced cicatricial conjunctivitis with symblephara. AB - PURPOSE: Cicatricial conjunctivitis is an uncommon ocular finding with several possible origins. We report a patient whose obsessive-compulsive disorder involved constant self-induced mechanical trauma to the eyes, leading to bilateral cicatricial conjunctivitis. METHOD: Case report. RESULTS: Based on the history, physical examination, ocular surface scraping, and conjuctival biopsy, other causes of cicatrical conjunctivitis were ruled out, and a self-induced cause was confirmed. CONCLUSIONS: Inquiries pertaining to psychiatric history may be important in patients with cicatricial conjunctivitis. PMID- 9220253 TI - Solitary corneal myxoma. AB - PURPOSE: To report the clinicopathologic findings of a myxoma that arose in the subepithelial region of the right cornea of a 53-year-old man 4 years after successful treatment of an infectious corneal ulcer. METHODS: Histopathologic and histochemical evaluation of corneal tissue. RESULTS: This rare lesion appears to have originated from corneal stromal fibroblasts that reacted to an inflammatory stimulus and produced excessive amounts of glycosaminoglycans (hyaluronic acid) rather than normal collagen. CONCLUSION: Myxoma formation may require interruption of Bowman's layer and proximity of the scar to the epithelium. PMID- 9220254 TI - Neural stimulation of lactoferrin and epidermal growth factor secretion by the lacrimal gland. PMID- 9220255 TI - The relationship between retrograde and anterograde amnesia in patients with typical global amnesia. AB - An extensive battery of tests of anterograde amnesia and remote memory was given to ten amnesics with lesions either to the medial temporal lobes of the diencephalon. These showed that the patients had anterograde amnesia with deficits in verbal and non-verbal recall and recognition, but preservation of word stem completion and intelligence. Mild impairments on executive tests and digit span performance were largely caused by the poor performance of the Korsakoff patients. The amnesics also showed remote memory deficits for personal and public domain information, and temporal gradients were observed for some of the tests. These deficits probably arose because the patients' anterograde amnesia was more severe than their retrograde amnesia even for the recent pre morbid past. They were more impaired in the recall of details about famous names in their ability to recognize such names. There was also a suggestion that performance on anterograde tests did not relate strongly to that on tests of retrograde amnesia of the remote pre-morbid past. However, this effect was less apparent with memory for personal information when the format and the information tapped were matched on pre- and post-morbid tests. PMID- 9220257 TI - Hand, space and attentional asymmetries in goal-directed manual aiming. AB - Two experiments were conducted to explore the interaction of the two cerebral hemispheres in motor control, by examining hand, space and attentional asymmetries in goal-directed aiming. In Experiment 1, right-handed subjects moved to targets more quickly with their right hand than their left hand. In addition, each hand was faster when moving in its own hemispace. Although in a control condition, movements were initiated more quickly with the left hand, visual distractors disrupted left hand performance more than right hand performance. For contralateral aiming, ipsilateral distractors caused the greatest interference. In Experiment 2, when targets and distractors were all presented at the midline, a right hand advantage was found for movement time along with a left hand advantage for reaction time, independent of target and distractor location. Our findings are discussed in terms of a right hemisphere role in movement preparation and the allocation of attention in space, and greater left hemisphere involvement in movement execution. PMID- 9220256 TI - Cerebral pathways for calculation: double dissociation between rote verbal and quantitative knowledge of arithmetic. AB - We describe two acalculic patients, one with a left subcortical lesion and the other with a right inferior parietal lesion and Gerstmann's syndrome. Both suffered from "pure anarithmetia": they could read arabic numerals and write them to dictation, but experienced a pronounced calculation deficit. On closer analysis, however, distinct deficits were found. The subcortical case suffered from a selective deficit of rote verbal knowledge, including but not limited to arithmetic tables, while her semantic knowledge of numerical quantities was intact. Conversely the inferior parietal case suffered from a category-specific impairment of quantitative numerical knowledge, particularly salient in subtraction and number bissection tasks, with preserved knowledge of rote arithmetic facts. This double dissociation suggests that numerical knowledge is processed in different formats within distinct cerebral pathways. We suggest that a left subcortical network contributes to the storage and retrieval of rote verbal arithmetic facts, while a bilateral inferior parietal network is dedicated to the mental manipulation of numerical quantities. PMID- 9220258 TI - Anatomical and neurological correlates of acute and chronic visuospatial neglect following right hemisphere stroke. AB - Anatomical and neurological correlates of visuospatial neglect were studied in 53 patients with a CT-documented right hemisphere stroke. Evidence of neglect at the acute stage poststroke was strongly related to large lesions involving the middle temporal gyrus and/or the temporo-parietal paraventricular white matter. Thus, out of 18 patients with evidence of visuospatial neglect at the acute stage, 12 showed a lesion in the middle temporal gyrus and/or the deep temporo-parietal white matter. Among the 35 patients that failed to show visuospatial neglect, only one patient had a lesion within these areas. Comparing those patients who recovered from neglect with those that did not, a high correlation was found between persisting neglect and a lesion involving the paraventricular white matter in the temporal lobe. On the basis of above findings, it was suggested that a simultaneous damage to the cortico-thalamic system for regulation of arousal and to the neural systems mediating visual orienting, is likely to be followed by persisting neglect symptoms. PMID- 9220259 TI - Alien hand syndrome: influence of neglect on the clinical presentation of frontal and callosal variants. AB - Three patients with mesial frontal and extensive callosal lesions due to anterior cerebral artery infarction manifested an alien hand syndrome (AHS) with varied features. Patient 1 with left hemispheric lesion showed right hand's impulsive reaching and grasping and left hand's antagonistic movements to the right (intermanual conflict; IMC). Patients 2 and 3 with right hemispheric lesion manifested a left hemihypokinesia which was thought to have suppressed the frequency and amplitude or even the occurrence of left hand's reaching and grasping. IMC and other left hand's non-antagonistic, irrelevant movements to the right remained. Because the term "IMC" is often misused and not strictly defined, its association with right hand's reaching and grasping is quite uncommon, its significance as a sign of callosal disconnection is not well validated, and because left hand's reaching and grasping tend to be suppressed by motor neglect, a trend may then develop for the right hand to be the sole focus of pathological behaviour in patients with the so-called frontal AHS (Feinberg, Schindler, Flanagan et al., 1992). PMID- 9220260 TI - What does the haptic modality do during cognitive activities on letter shapes? A study with left- and right-handers. AB - This study was undertaken to analyze intermanual (interhemispheric) transfer in left and right handed subject and to assess how information was extracted during finger scanning of letter shape at the different levels of letter processing: shape recognition during a physical matching task, letter recognition in a verbal "meaning" matching task and letter naming. The dichhaptic procedure was used to study interhemispheric relations. It was hypothesized that cognitive activities have a feed-forward effect on the exploration of shapes, and that the performance is related to the nature of the task and to handedness. The exploratory strategies of the two types of handedness were also analyzed. The results showed that response latencies were generally similar for left- and right-handed subjects, but accuracy was better for left than right handers in "verbal" matching with the same overall exploratory strategies. In physical matching, left and right-handed subjects performed equally but used different exploratory strategies. The naming task was very difficult for both groups but failed to discriminate their on accuracy, response latency, and exploratory strategy. The results are discussed with reference to the different exploratory strategies used and the interhemispheric interaction at work in different cognitive processes. PMID- 9220261 TI - Unilateral neglect and space constancy during passive locomotion. AB - Space constancy was investigated in seven blindfolded left-neglect patients by driving them along routes involving one or two, left or right, 90 degrees turns. At the end of each route patients had to indicate its starting point while still blindfolded. On average, no considerable left/right differences were found in pointing accuracy. The entailments of this finding for the understanding of neglect phenomena are briefly discussed. PMID- 9220262 TI - The novelty effect in recovered hemineglect. AB - Left neglect patients, patients who had recovered from left neglect and control subjects performed a task of simple motor reaction times (RTs) to lateralised visual stimuli. Neglect and recovered patients were slower than controls on left sided targets. To explore the time course of the allocation of attention across space, an analysis of responses as a function of the serial order of the trials was performed. While neglect patients' performance did not substantially change over time, recovered patients showed a stereotyped 'novelty effect', consisting of larger left/right RT differences at the beginning of the task than at the end of it. To explain this practice-related change, a trade-off is hypothesised between the process of learning the motor task and the mechanisms involved in recovery from neglect, such as the reorienting of attention toward the contralesional side following the initial ipsilesional orienting. A possible role is proposed for the prefrontal cortex as the crucial neural structure that mediates both processes. PMID- 9220263 TI - Access to information about famous individuals in Alzheimer's disease. AB - Alzheimer's disease (AD) patients and normal adults were tested in two complementary recognition tasks. On each trial of the faces task, participants matched the photograph of a famous individual to one of four names. On each tial of the names task, participants matched the name of a famous individual to one of four photographs. The AD patients made enough consistent errors across the two tasks to suggest an impairment in the storage of information about the individuals. In addition, they made enough inconsistent errors to suggest a generalized retrieval deficit. The AD patients performed as well on the faces taslk as on the names task, providing no evidence of a specialized deficit in the retrieval of lexical information. PMID- 9220264 TI - On crossed apraxia. Description of a right-handed apraxic patient with right supplementary motor area damage. AB - GP, a right-handed woman, without evidence of familial left-handedness, showed clearcut bilateral ideo-motor apraxia and oro-facial apraxia after a vascular lesion of the right hemisphere, encroaching upon the fronto-mesial region. She scored normally in most other cognitive tests, including language, but showed signs of callosal disconnection, left anarchic hand and mild unilateral spatial neglect. This cognitive profile points to the possibility of praxis being localized to the right hemisphere in this right-handed patient. We argue in favour of individual variability of praxis dominance, and maintain that this dominance might be completely right-sided in some subjects. Moreover the anatomical locus of GP's lesion points to the possible role that the frontal lobes (and more specifically the Supplementary Motor Area) play in the genesis of apraxia. PMID- 9220265 TI - Acquired dysgraphia in alphabetic and stenographic handwriting. AB - We report the unusual case of AZO, who professionally used handwritten shorthand writing, and became dysgraphic after a stroke. AZO suffered from a complex cognitive impairment, and part of her spelling errors resulted from damage to auditory input processing, to phonology-orthography conversion procedures and to the ortographic output lexicon. However, analysis of her writing performance showed that the same variables affected response accuracy in alphabetic and shorthand writing; and, that the same error types, including transpositions, were observed in all tasks in the two types of writing. These observations are consistent with damage to the graphemic buffer. They suggest that, in multiple code writing systems (e.g., stenography, Japanese, or in the case of multilingual speakers of languages that use different spelling codes), the graphemic buffer is shared by all codes. PMID- 9220266 TI - Preserved imagery for colours in a patient with cerebral achromatopsia. AB - We report the case of a patient who, after sequential bilateral strokes in the occipital regions sparing the primary visual cortex, developed a severe deficit of colour perception. At variance with other reports of acquired achromatopsic patients, she showed a perfectly vivid visual imagery for colours. These findings, together with similar data in domains other than colour processing, challenge the theories which posit that the same cognitive processes are involved in both the perception and the retrieval from memory of a given stimulus. PMID- 9220267 TI - Personal versus extrapersonal neglect: a group study of their dissociation using a reliable clinical test. AB - The validation of a simple quantitative clinical test of personal neglect is described in this study of 17 right brain damaged CVA patients with extrapersonal neglect, 14 without unilateral extrapersonal neglect, 13 left brain damaged CVA patients and 17 age-matched controls. The test had a high reliability and clearly differentiated neglect patients from all other groups. Furthermore the test identified a much higher incidence of personal neglect among extrapersonal neglect patients (59%) than has previously been found. Moreover this study confirms earlier findings by showing a double dissociation between personal and extrapersonal neglect. Seven patients with extrapersonal neglect showed no personal neglect while five patients showing no extrapersonal neglect did show personal neglect on this test. PMID- 9220268 TI - Deficits in delayed memory following cerebral malaria: a case study. AB - Cerebral malaria is a common disease, but there have not been any reports or investigations of long-term neurological or neuropsychological outcome. We present a case in which severe deficits in delayed memory and naming ability are observed 10 years after the patient contracted cerebral malaria. Neuropsychological testing and medical imaging are both consistent with temporal lobe/hippocampal dysfunction, which corroborates earlier animal research that cerebral malaria is particularly likely to lead to interrupted blood circulation in this area. PMID- 9220269 TI - Effects of calcium antagonists on the risks of coronary heart disease, cancer and bleeding. Ad Hoc Subcommittee of the Liaison Committee of the World Health Organisation and the International Society of Hypertension. PMID- 9220270 TI - The kidney and hypertension. AB - The kidney is important not only as a target organ of hypertension but also as a organ that may cause hypertension. For many years, we have studied basic and clinical aspects of the role of the kidney in hypertension, including the renin angiotensin, kallikrein-kinin, and prostaglandin systems, glomerular hemodynamics, therapeutic modalities, and prognosis. This review summarizes some of our results together with those of other relevant studies in the literature. PMID- 9220271 TI - Left ventricular hypertrophy is more prominent in patients with primary aldosteronism than in patients with other types of secondary hypertension. AB - We determined functional and morphological changes of the heart by 2-dimensional and pulse Doppler echocardiography in 20 patients with primary aldosteronism and compared the results with those in 50 healthy normotensive subjects, 12 patients with Cushing's syndrome, 9 patients with pheochromocytoma, and 47 patients with essential hypertension. All hypertensive groups had greater left ventricular mass indexes than did the normotensive group (76.9 +/- 17.2 g/m2). Despite similar age distribution, blood pressure during antihypertensive treatment, and duration of hypertension, the primary aldosteronism group had a significantly greater left ventricular mass index (152.5 +/- 42.5 g/m2) than did the Cushing's syndrome (103.4 +/- 37.5 g/m2), pheochromocytoma (122.4 +/- 28.5 g/m2), and essential hypertension (101.4 +/- 32.8 g/m2) groups. The left ventricular posterior wall thickness and interventricular septal wall thickness were significantly greater in the hypertensive groups than in the normotensive group and also significantly greater in the primary aldosteronism group than in any of the other hypertensive groups. By contrast, there were no significant differences among the four hypertensive groups in any variable of systolic or diastolic function of the heart. The results suggest that left ventricular hypertrophy is more pronounced in patients with primary aldosteronism than in patients with other forms of hypertension. It is therefore important to echocardiographically evaluate cardiac hypertrophy as a risk factor of morbidity and mortality in patients with this low renin hypertension. PMID- 9220272 TI - Immunosuppressant HR-325 attenuates progression of malignant arteritis in the kidney of Dahl salt-sensitive rats. AB - We investigated the effects of the immunosuppressant HR-325 on arterial lesions in Dahl rats with salt-induced hypertension. Forty-eight 6-wk-old Dahl salt sensitive (DS) rats were divided into 1) a low-salt (0.3% NaCl) group, 2) a high salt (4% NaCl) group, 3) a high-salt and low-dose (1 mg/kg) HR-325 group, and 4) a high-salt and high-dose (30 mg/kg) HR-325 group. The rats were treated for 8 wk. Various variables of renal function and morphological alterations in the kidney were assessed. Blood pressure was measured by the tail-cuff method. HR-325 significantly decreased systolic blood pressure in a dose-dependent manner throughout the study. HR-325 tended to decrease plasma creatinine level and increase creatinine clearance rate. Morphological studies revealed that HR-325 treatment strikingly resolved infiltration of immune-related cells in perivascular and intraluminal lesions, thereby decreasing the total arterial injury score by 32%. High-dose HR-325 also attenuated glomerulosclerosis and tubular injury by 35% and 34%, respectively, as compared with untreated high-salt Dahl S rats. Reduced levels of immune-related cells resulted in a decrease in urinary nitrite excretion. These data indicate that long-term treatment with the immunosuppressant HR-325 decreases systolic blood pressure in Dahl salt-sensitive rats, and that this decrease is associated particularly with resolution of infiltration of immune-related cells in arterial lesions. Hyperimmune state is responsible in part for the susceptibility of Dahl S rats to hypertensive organ damage. PMID- 9220273 TI - Personality characteristics of patients with "white coat" hypertension. AB - To clarify psychological factors related to white coat hypertension, we examined personality characteristics of patients with mild essential hypertension by psychological testing. Patients with essential hypertension were taught to measure their own blood pressure (BP) with a semi-automatic oscillometric BP measuring device and were asked to measure BP at home in the sitting position before going to sleep. The duration of the study was 8 wk. Patients were defined as "white coat" hypertensive patients (WCHT) (n = 49) if home systolic BP was 135 mmHg or less and home diastolic BP was 85 mmHg or less, and as "sustained" hypertensive patients (SHT) (n = 53) if home systolic BP was 140 mmHg or more or home diastolic BP was 90 mmHg or more. All the patients underwent the following psychometric tests: self-rating questionnaire for depression, MMPI alexithymia scale, type A behavior pattern check list, general health questionnaire (GHQ), and egogram check list. WCHT did not differ from SHT in the scores for depression, alexithymia, type A behavior pattern, or GHQ. However, WCHT showed an abnormal pattern on egograms, as compared with SHT. On egograms, SHT showed a normal hill-shaped pattern with a peak in "nurturing parent (NP)", and "free child (FC)" was higher than "adapted child (AC)" in both genders. In contrast, WCHT showed significantly lower FC and significantly higher AC than SHT, and AC was higher than FC in both genders. These findings suggested that WCHT tend to suppress their own emotions and become over-adaptive to their surroundings, as compared with SHT. PMID- 9220274 TI - Effects of long-term antihypertensive therapy on physical fitness of men with mild hypertension. AB - This study was conducted to investigate the effects of long-term administration of a calcium-channel antagonist (nifedipine) and a beta-blocker (acebutolol) on physical fitness in men with mild hypertension. All subjects underwent symptom limited treadmill stress testing and routine echocardiographic studies. Twenty two subjects who had either a causal diastolic blood pressure of more than 105 mmHg or a left ventricular mass index (LVMI) of 125 g/m2 or more during follow-up were assigned to receive medical therapy. The other 31 men who did not meet either criterion were continuously followed-up without medication. Among the 22 treated men, the age-adjusted treadmill time (normalized treadmill time, TMTn) significantly decreased before the initiation of medication, while 31 untreated men showed no change in TMTn throughout the study. The 22 treated subjects were subsequently divided into two groups; 13 were given nifedipine and 9 were given acebutolol. All treated subjects were followed-up for more than 3 years. After treatment, the two groups showed similar reductions in blood pressure and LVMI, but a different outcome for TMTn: TMTn increased from 104 +/- 8% to 115 +/- 16% in subjects given nifedipine (p < 0.05) and decreased from 106 +/- 12% to 99 +/- 10% (p < 0.01) in those given acebutolol. Thus, the physical fitness of subjects who required medication significantly deteriorated without medication; their physical fitness improved after treatment with a calcium-channel antagonist and deteriorated after treatment with a beta-blocker. PMID- 9220275 TI - Changes in cardiac adrenomedullin concentration in renovascular hypertensive rats. AB - We assessed changes in tissue and plasma adrenomedullin levels in two-kidney, one clip renovascular hypertensive rats. Four weeks after clipping, adrenomedullin concentrations were significantly higher in the cardiac ventricles and lower in the left atrium than the respective values in sham-operated rats. The left ventricular adrenomedullin concentration significantly correlated with systolic blood pressure and the degree of cardiac hypertrophy. No difference was noted in the adrenomedullin concentrations of the adrenal gland, aorta, lung, kidneys, or plasma between the two groups. These findings indicate possible involvement of cardiac adrenomedullin in this model of hypertension. PMID- 9220276 TI - Prejunctional regulation by endogenous and exogenous acetylcholine of adrenergic nerve function in isolated canine mesenteric arteries. AB - Transmural electrical stimulation (5-30 Hz) produced a frequency-dependent increase in the perfusion pressure of isolated, perfused dog mesenteric artery segments without the endothelium, which was abolished by prazosin or tetrodotoxin. Physostigmine inhibited the pressor response to transmural electrical stimulation, whereas atropine potentiated the response. Treatment with acetylcholine (10(-6) and 10(-5) M) dose-dependently inhibited the response to electrical nerve stimulation. The effect was reversed by the addition of atropine and AF-DX 116 at a concentration (10(-7) M) that selectively blocked the M2 receptor subtype, but not by pirenzepine or 4-DAMP. Acetylcholine did not alter the pressure raised by norepinephrine in perfused arterial segments nor the contraction caused by exogenous norepinephrine in the artery strips. 3H-overflow evoked by transmural electrical stimulation from tissues prelabeled with [3H] norepinephrine was decreased by acetylcholine (10(-6) M) in the superfused dog mesenteric arterial strips. It is concluded that acetylcholine inhibits adrenergic neurogenic contractions by interfering with the release of norepinephrine, which possibly results from activation of the prejunctional M2 receptor subtype. PMID- 9220277 TI - Influence of aging on progression of cardiovascular complications associated with insulin resistance in patients with essential hypertension. AB - Hyperinsulinemia or insulin resistance is suggested to play a role in the pathogenesis of hypertension and its target organ diseases. It is also well documented that aging is associated with a decline in glucose tolerance and insulin sensitivity, but there are few reports on the relationship between aging and insulin sensitivity or on the effects of aging on the progression of cardiovascular complications in patients with essential hypertension. To clarify these effects of aging in essential hypertension, 44 patients were examined by the euglycemic hyperinsulinemic glucose clamp test and ultrasonography of the heart and carotid arteries. There was a significant negative correlation between aging and insulin sensitivity (r = -0.37, p < 0.05). Significant increases in left ventricular mass index and carotid wall thickening accompanied by insulin resistance were seen in only non-elderly patients but not in elderly patients. These results suggest that aging decreases insulin sensitivity even in essential hypertensive subjects and that insulin resistance does not affect the progression of cardiac hypertrophy and atherosclerosis in elderly patients with essential hypertension. PMID- 9220278 TI - Effects of angiotensin AT1 receptor antagonist on volume overload-induced cardiac gene expression in rats. AB - The present study was undertaken to examine the effects of volume overload on cardiac gene expression and the possible role of angiotensin AT1 receptor in such expression. Cardiac volume overload was prepared by abdominal aortocaval shunt in rats. Rats with aortocaval shunt were treated with 1) vehicle, 2) an angiotensin AT1 receptor antagonist, CS-866 (10 mg/kg/d), or 3) an angiotensin-converting enzyme inhibitor, temocapril (10 mg/kg/d), for 7 days. Cardiac tissue mRNA was measured by Northern blot analysis with specific probes. Aortocaval shunt not only caused cardiac hypertrophy but also upregulated the gene expression of atrial natriuretic polypeptide, collagen III, and downregulated Ca(2+)-ATPase expression in the left ventricle. These changes were prevented by treatment with CS-866, while temocapril failed to normalize left ventricular Ca(2+)-ATPase expression. Unlike the left ventricle, the significant downregulation of alpha myosin heavy chain and transforming growth factor-beta 3 by aortocaval shunt was observed in the right ventricle, and CS-866 normalized this decreased expression of transforming growth factor-beta 3. The left and right atria showed increased expression of collagen type I as well as of collagen type III and atrial natriuretic polypeptide, and these increases were more effectively prevented by CS-866 than by temocapril. Thus, the effects of cardiac volume overload on cardiac performance-related gene expression differ between the ventricles and atria. Our results suggest that AT1 receptor partially contributed to volume overload-induced changes in cardiac gene expression and that AT1 receptor antagonists and angiotensin-converting enzyme inhibitors have different effects in this model of cardiac hypertrophy. PMID- 9220279 TI - Indirect assessment of glomerular capillary pressure from pressure-natriuresis relationship: comparison with direct measurements reported in rats. AB - It is examined whether glomerular hemodynamics can be indirectly estimated from the pressure-natriuresis relationship. There are only two animal studies reported, one in normal and the other in 5/6 nephrectomized Munick-Wistar rats, which permits plotting the pressure-natriuresis relationship and comparison of indirect estimations with directly measured glomerular hemodynamic data. Normal and extensive renal ablation rats were placed on relatively high and low sodium diets. Plotting mean arterial pressure (MAP) on the x-axis and 24 h urinary sodium excretion rate on the y-axis, the pressure-natriuresis relationship was drawn. As the difference between MAP (121 +/- 1 and 169 +/- 12 mmHg) on relatively high sodium diet and the extrapolated x-intercept (122 and 138 mmHg) of the pressure-natriuresis relationship, based on previous proposal, the effective filtration pressure across the glomerular capillary walls was estimated to be-1 and 31 mmHg for normal and 5/6 nephrectomized rats. Then, the glomerular capillary hydraulic pressure (PGC) was calculated to be 33 and 64 mmHg. Micropuncture studies showed that directly measured PGC of 47 +/- 1 and 65 +/- 2 mmHg was close agreement with those estimated indirectly. Therefore, an approach from the pressure-natriuresis relationship provides a noninvasive means to predict an approximation of PGC. This approach to estimating glomerular hemodynamics may have invaluable implications for clinical practice; in the early detection of loss of filtration capacity and in the assessment of an important risk factor for the development of chronic renal failure. PMID- 9220281 TI - MRI of coronary arteries. AB - Magnetic Resonance Angiography (MRA) of the coronary arteries has recently become possible due to the development of a new group of ultrafast imaging sequences. Although the role of coronary MR angiography in screening for coronary artery lesions has not yet been established, coronary MR angiography has been very successful in the detection of coronary artery variants, and the imaging of coronary stents and bypass grafts. Variants of these new MRI techniques can also quantitate velocity in native coronary arteries. Coronary MR angiographic techniques can be subdivided in breath-hold (single or repeated breath-hold) and non-breath-hold techniques. Most of the clinical experience so far has been with a single breath-hold technique, and was limited to cooperative patients. The recent introduction of navigator pulses for real-time respiratory gating or triggering allows non-breath-hold or repeated breath-hold 3-D coronary MR angiography, and will allow a more widespread use of this technique. Notwithstanding the progress being made and the excitement created by the prospect of a noninvasive coronary artery screening tool, several key technical problems remain unresolved and are now being addressed by the scientific and clinical community. This paper reviews ongoing research in coronary MR angiography. PMID- 9220280 TI - Ischemic heart disease: value of MR techniques. AB - BACKGROUND: The cardiovascular applications of magnetic resonance (MR) techniques in coronary artery disease have increased considerably in recent years. Technical advantages of MR imaging are the excellent spatial resolution, the characterization of myocardial tissue, and the potential for three-dimensional imaging. These characteristics allow the accurate assessment of left ventricular mass and volume, the differentiation of infarcted from normal tissue, and the determination of systolic wall thickening and regional wall motion abnormalities. METHODS: In addition to the conventionally used spin-echo and cine-echo techniques, newer techniques such as myocardial tagging, ultrafast MR imaging and MR coronary angiography have been developed. These newer techniques allow a more accurate assessment of ventricular function (tagging), myocardial perfusion (ultrafast imaging), and evaluation of stenosis severity (MR coronary angiography). Particularly early detection and flow assessment of stenosed coronary arteries and bypasses by MR angiography would constitute a major breakthrough in cardiovascular MR imaging. Apart from the MR imaging techniques, cardiac metabolism may be well assessed using MR spectroscopy. This provides unique information on the metabolic behaviour of the myocardium under conditions stress-induced ischemia. However, the definite niche of cardiac MR spectroscopy has still to be settled. CONCLUSION: Currently, MR techniques allow the evaluation of anatomy and function (accepted use), perfusion and viability (development phase), and coronary angiography (experimental phase). A particular strength of MR imaging is that one single MR test may encompass cardiac anatomy, perfusion, function, metabolism and coronary angiography. The replacement of multiple diagnostic tests with one MR test may have major effects on cardiovascular healthcare economics and would outweight the cost inherent to the MR angiography procedure. PMID- 9220282 TI - A practical approach to MRI of coronary artery bypass graft patency and flow. AB - Direct visualization of coronary artery bypass grafts can be obtained non invasively by magnetic resonance imaging. Several studies demonstrated a high sensitivity and somewhat lower specificity for detection of vein-graft patency, using the conventional spin-echo and gradient-echo techniques. In addition, the true functional status can be assessed by determining the flowrate within the graft using phase velocity mapping. Important limitations of the previously applied techniques include the inability to accurately evaluate the different segments of jump grafts and the presence of graft stenoses. Further improvement is to be expected from the recent introduction of breath-hold imaging sequences and the forthcoming introduction of bloodpool-avid contrast agents. PMID- 9220283 TI - MR first pass imaging: quantitative assessment of transmural perfusion and collateral flow. AB - Recent advances with fast switching gradient coils, and the optimization of magnetic resonance techniques for multislice imaging have made it possible to apply models of contrast agent transit for the quantification of myocardial perfusion, and determination of the transmural distribution of blood flow. This article summarizes some of these recent developments and presents examples of quantitative, multi-slice myocardial perfusion imaging studies in patients and animal models. Multi-slice, true first pass imaging, with high temporal resolution, and T1-weighted, arrhythmia insensitive contrast enhancement is used for the quantification of perfusion changes accompanying mild to severe ischemia. The first pass imaging technique and the modeling approach are sufficiently robust for fitting of tissue residue curves corresponding to a wide, physiologically realistic range of myocardial blood flows. In animals this was validated by comparison to blood flow measurements with radiolabeled microspheres as gold standard. It is demonstrated that with the proposed modeling approach one can determine the myocardial perfusion reserve from two consecutive MR first pass measurements under resting and hyperemic conditions. In patients with microvascular dysfunction the MR studies show for the first time that the myocardial perfusion reserve correlates with Doppler flow measurements (linear regression with slope of 1.02 +/- 0.09; r = 0.80). Since perfusion limitations usually begin in the subendocardium as coronary flow is gradually reduced, first pass imaging with the prerequisitie spatial and temporal resolution allows early detection of a mild coronary stenosis. PMID- 9220285 TI - Left ventricular myocardial tagging. AB - Nuclear Magnetic Resonance myocardial tagging is a potent non-invasive technique which enables the quantification of myocardial deformation, globally but also regionally at different time points during the cardiac cycle. By the use of presaturating pulses prior to the actual imaging sequence non-invasive markers or tags can be placed on the myocardium at end diastole, which move and deform with the underlying myocardium on which they were inscribed. Through combination of perpendicular sets of short- and long-axis images with tags, a three dimensionally reconstructed left ventricle is obtained, subdivided in 32 myocardial cuboids for which the 3D coordinates of the corners are known at different time points in the cardiac cycle. From these data global and regional strains and quantitative measures of shape can be computed. PMID- 9220284 TI - Magnetic resonance imaging in valvular heart disease. AB - Magnetic resonance techniques can be employed to depict valvular abnormalities but are especially helpful in quantifying regurgitant or stenotic lesions which cannot be quantitatively assessed by other noninvasive techniques. Gradient echo techniques and phase velocity mapping are the most important magnetic resonance pulse sequences employed for these purposes. Valvular regurgitation can be quantitated by measuring the area of signal void on conventional gradient-echo images, by calculating stroke volume differences from k-space segmented gradient echo images, by measuring the proximal convergence zone from velocity encoded images or by comparing stroke volumes of the ventricles from velocity measurements. In contrast to this variety of possibilities in regurgitant lesions, stenotic lesions can only be quantitated by using velocity mapping techniques. Magnetic resonance spectroscopy can be used to assess myocardial metabolism in chronic valvular lesions. However, this tool needs further development and more clinical data before its use can be recommended to assess the necessity and optimal timing of surgical intervention. PMID- 9220286 TI - Quantitative analysis of cardiovascular MR images. AB - The diagnosis of cardiovascular disease requires the precise assessment of both morphology and function. Nearly all aspects of cardiovascular function and flow can be quantified nowadays with fast magnetic resonance (MR) imaging techniques. Conventional and breath-hold cine MR imaging allow the precise and highly reproducible assessment of global and regional left ventricular function. During the same examination, velocity encoded cine (VEC) MR imaging provides measurements of blood flow in the heart and great vessels. Quantitative image analysis often still relies on manual tracing of contours in the images. Reliable automated or semi-automated image analysis software would be very helpful to overcome the limitations associated with the manual and tedious processing of the images. Recent progress in MR imaging of the coronary arteries and myocardial perfusion imaging with contrast media, along with the further development of faster imaging sequences, suggest that MR imaging could evolve into a single technique ('one stop shop') for the evaluation of many aspects of heart disease. As a result, it is very likely that the need for automated image segmentation and analysis software algorithms will further increase. In this paper the developments directed towards the automated image analysis and semi-automated contour detection for cardiovascular MR imaging are presented. PMID- 9220287 TI - Pre-clinical activity of taxol in non-seminomatous germ cell tumor cell lines and nude mouse xenografts. AB - Taxol (Paclitaxel) is a novel anti-cancer drug which has shown excellent clinical activity in a variety of solid tumors, particularly in metastatic breast and ovarian cancer. 70-80% of patients with metastatic non-seminomatous germ cell tumor (NSGCT) attain disease-free status with standard cisplatin-based combination chemotherapy but the emergence of drug resistance still prevents a small proportion of these patients from achieving long-term remission. Here we report the results of pre-clinical studies investigating whether taxol exhibits cross-resistance to cisplatin or ifosfamide in human NSGCT cell lines and in a cisplatin refractory xenograft model of human NSGCT. Following 96-h drug exposure in a 5-day sulphohodamine B (SRB) in vitro assay, taxol demonstrated potent cytotoxicity in cell lines which were cisplatin sensitive (577 LM, H32, H12.1; mean IC50s 1.5-3.0 nM) or those with acquired or intrinsic cisplatin resistance (H12DDP, H23.1; mean IC50s 2.5 nM). Compared to the drug-sensitive cell line, H12.1, the IC50 values of taxol were increased in cell line 1777NRp Cl-A with intermediate level resistance to cisplatin and ifosfamide (4.7 nM; p > 0.05) and significantly elevated in cell line 1411HP, with a high level of cisplatin resistance (6.9 nM; p < 0.01). The latter 2 cell lines may represent models corresponding to patients relapsing after high-dose platinum-based chemotherapy who seem to be resistant to taxol therapy. The IC50s of taxol in H32 and H12DDP were approximately 100-fold lower following drug exposure times exceeding 24 hours compared with short exposure times (1-6 h). Dose-dependent anti-tumor activity was observed with taxol in a cisplatin-refractory xenograft model of NSGCT (H23.1), with significant anti-tumor activity observed at a dose of 15 mg/kg/d injected intravenously on days 1 through 5. The results of this study are in accordance with the most recent clinical data which showed that taxol is a useful drug in relapsed or cisplatin-refractory testicular germ cell cancer, with significant anti-tumor activity being observed in 25% of patients, but poor activity in patients previously treated with high-dose therapy. Further pre clinical research, especially using models such as 1411HP and 1777NRp Cl-A, on the combinations of taxol with other regimens are required to enable successful treatment of the most drug-resistant relapsed germ cell tumors. PMID- 9220288 TI - Treatment of human prostate tumors PC-3 and TSU-PR1 with standard and investigational agents in SCID mice. AB - Both the PC-3 and the TSU-PR1 prostate tumor models were found to be satisfactory for chemotherapeutic investigations in ICR-SCID mice. The 30 to 60 mg fragments implanted took in all mice (as judged by 100% takes in the controls of all experiments as well as the passage mice). The tumor volume doubling time was 4.0 days for PC-3 and 2.5 days for TSU-PR1. Nine agents were evaluated IV against early stage subcutaneous PC-3 tumors, with Nano-piposulfan being the only agent highly active (4.9 log kill). Three other agents were moderately active: Taxol (1.5 log kill), Cryptophycin-8 (1.6 log kill), Vinblastine (1.0 log kill). Five agents were inactive: VP-16, Adriamycin, CisDDPt, 5-FUra, and Cyclophosphamide. Ten agents were evaluated IV against early stage subcutaneous TSU-PR1 tumors. Three agents were highly active, producing > 6 log kill and cures: Taxol (5/5 cures), Cryptophycin-8 (5/5 cures), Vinblastine (2/4 cures). Two other agents were moderately active: Nano-piposulfan (1.2 log kill), and Cyclophosphamide (1.1 log kill). Five agents were inactive: VP-16, Adriamycin, CisDDPt, 5-FUra, and BCNU. In part, activity was determined by the ability of the SCID mice to tolerate meaningful dosages of the agents. Agents producing granulocyte toxicity (e.g., Adriamycin) were poorly tolerated and appeared less active than expected. Vinblastine, producing little or no granulocyte toxicity was very well tolerated and appeared to be more active than expected. PMID- 9220289 TI - Comparative cytotoxicity of oxaliplatin and cisplatin in non-seminomatous germ cell cancer cell lines. AB - Approximately 70-80% of patients with metastatic testicular cancer will become disease free with cisplatin-based chemotherapy and most of these patients will be long-term survivors. Despite these impressive results, the two limitations of cisplatin are its severe and potentially long-term side-effects, and the emergence of drug resistance which prevents a small proportion of these patients from achieving long-term remission. Oxaliplatin has an improved toxicity profile compared to cisplatin and contains the diaminocyclohexane (DACH) substituent known to be correlated with a lack of cross-resistance with cisplatin. A phase II study has shown interesting activity when used in combination with cisplatin in cisplatin-refractory testicular cancer patients. Here we report the results of the first in vitro study investigating whether oxaliplatin as a single agent exhibits cross-resistance to cisplatin in a panel of non-seminomatous germ cell tumor (NSGCT) cell lines using short and long-term drug exposures in a five-day sulfhodamine B in vitro cytotoxicity assay. Oxaliplatin cytotoxicity was significantly superior to cisplatin in cell lines with both acquired (H12DDP) and intrinsic (1777NRp Cl-A) intermediate level resistance to cisplatin. Following 24 h or 96 h drug exposure the fold resistance in H12DDP and 1777NRp Cl-A was 1.7 2.2 with oxaliplatin compared to 3.9-6.1 with cisplatin. The cytotoxic activity of oxaliplatin was not significantly different from that of cisplatin in cisplatin-sensitive cell lines or in cell lines with a high level (10-20 fold) of cisplatin resistance. The results of this study suggest that further preclinical studies in NSGCT are of interest, particularly in combination with cisplatin, ifosfamide and etoposide. Furthermore, the in vitro results support the use of an oxaliplatin administration schedule giving prolonged drug exposure, such as the flat or circadian rhythm-modulated schedule already under investigation for oxaliplatin. PMID- 9220290 TI - Prolonged infusion gemcitabine: a clinical phase I study at low- (300 mg/m2) and high-dose (875 mg/m2) levels. AB - Gemcitabine (GEM) is a novel nucleoside analogue with a unique mechanism of action. Preliminary studies have shown a mild, schedule-dependent toxic profile with a broad range of MTDs and promising antitumor activity in various solid tumors. This phase I study describes the infusion length-effect relationships of low- (300 mg/m2) and high-dose (875 mg/m2) GEM, administered on days 1, 8 and 15 at 4-week intervals in a step-wise escalation of duration (> or = 33%) at a starting level of 60 minutes. At least 3 patients entered each infusion-level step and 3 more cases were treated in the presence of significant toxicity. Conservative criteria for toxicity were employed, including treatment delay until recovery with infusion de-escalation in the subsequent course. Forty-seven patients (29 at low- and 18 at high-dose GEM levels) with various solid tumors, including 9 (taken as a reference) who had received the same dose levels over 30 min. entered the study. All but 9 patients (with pancreatic cancer) had been previously treated with chemotherapy and all had extensive visceral disease. A striking infusional-effect relationship was observed at both GEM dose levels. Four escalation steps were required to define the maximum tolerated infusion time (MTIT) at 6 hours for 300 mg/m2 GEM, with leucopenia being dose-limiting. At 875 mg/m2, although no limiting toxicity was observed (in spite of increased severity of leucopenia), no escalation was attempted following the 1-hour infusion, due to the limiting rate (58% of 12 patients) of toxic delay requiring shorter infusions. Toxicity was usually mild (no grade 4 event was recorded) showing the usual profile, although there was a trend towards increased non-hematologic toxicity (i.e. LFT abnormalities) as compared with the MTD previously defined using a 30-min. infusion schedule (1,370 mg/m2). Eight patients achieved a PR: 1 with NSCLC, 1 with gastric and 2 with bladder cancer at 300 mg/m2 (1 with a 3- and 3 with a 6-hour infusion) and 2 with pancreatic, 1 with cervical and another with bladder cancer at 875 mg/m2 (all but one with a 1-hour infusion). These data clearly suggest that the infusion duration is an important independent factor that influences the clinical effects of GEM. The present study not only defined the toxic profiles and the MTITs of the selected dose levels but demonstrated that GEM retained the antitumor activity at doses as small as 300 mg/m2 when given as a prolonged infusion. Further studies should clarify the underlying mechanism(s) responsible for the erratic dose-effect dose-effect relationships of GEM and establish the optimal dose-infusion level in the treatment of solid tumors. PMID- 9220291 TI - Phase I trial of uracil-tegafur (UFT) plus oral leucovorin: 14-day schedule. AB - We previously reported results of a Phase II trial of UFT [Taiho Pharmaceutical Ltd., Tokyo, Japan; (BMS-200604) Bristol-Myers Squibb, Princeton, NJ], an oral 4:1 molar concentration of uracil and tegafur, plus oral leucovorin for metastatic colorectal carcinoma (Pazdur et al., J. Clin. Oncol. 12:2296-2300, 1994]. Our results demonstrated that a 28-day schedule of this combination produced a response rate similar to that obtained with conventional intravenous fluorouracil (5-FU)-plus-leucovorin regimens but without the severe or life threatening neutropenia or oral mucositis that complicates intravenous 5-FU regimens. The current Phase I trial examines the dose-limiting toxic effects and maximum tolerated dose of a 14-consecutive-day schedule of UFT plus oral leucovorin in 14 patients who had histologically proven cancer and had received prior chemotherapy. The daily UFT plus leucovorin dose was divided into three doses administered orally every 8 hours. In this study, the UFT dose was escalated while the leucovorin dose remained at 150 mg/day. Of the 14 patients, 4 were initially treated at the 350-mg/m2/day UFT level for 14 days without any dose-limiting toxic reactions. Subsequently, another 7 patients were treated at the 400-mg/m2/day level; grade 3 diarrhea developed in 3 of these 7 (with severe abdominal cramping in 2 cases and severe nausea and vomiting unresponsive to antiemetics in the third). To better define the starting dose for phase II studies, an additional 3 patients were treated at the 350-mg/m2/day dose level. Of the total 7 patients treated at 350 mg/m2/day, grade 3 toxic events (diarrhea) developed in 2 patients. Grade 1-2 toxic effects noted at this level included fatigue, stomatitis, skin rash, abdominal pain, nausea, and vomiting. Neither partial nor complete responses were observed in this trial. The maximum tolerated dose of this schedule is 350 mg/m2/day UFT plus 150 mg/day oral leucovorin. However, because of this schedule's inferior dose intensity compared with that of the 28-day schedule of UFT plus leucovorin, subsequent development of UFT in the United States has focused on the 28-day regimen. PMID- 9220292 TI - Phase I study of paclitaxel on a 3-hour schedule followed by carboplatin in untreated patients with stage IV non-small cell lung cancer. AB - This study sought to determine the principal toxicities and feasibility of administering paclitaxel as a 3-hour infusion followed by carboplatin without and with granulocyte colony-stimulating factor (G-CSF) in chemotherapy-naive patients with stage IV non-small cell lung carcinoma (NSCLC), and to recommend doses for subsequent clinical trials. Twenty-three patients were treated with paclitaxel at doses ranging from 175 to 225 mg/m2 followed by carboplatin targeting area under the concentration-time curve (AUC) 7 or 9 mg/mL.min every 3 weeks. AUCs were targeted using the Calvert formula with estimated creatinine clearance as a surrogate for the glomerular filtration rate. A high rate of intolerable, mutually exclusive toxicities, consisting primarily of thrombocytopenia, as well as neutropenia, nausea and vomiting, and mucositis, precluded escalation of carboplatin above a targeted AUC of 7 mg/mL.min with paclitaxel 225 mg/m2, which approaches the maximum tolerated dose (MTD) of paclitaxel given as a single agent on a 3-hour schedule. Moderate to severe peripheral neurotoxicity occurred in several patients after multiple courses. Due to the heterogeneous nature of the principal toxicities and the ability to administer clinically-relevant doses of both agents in combination without G-CSF, further dose escalation using G-CSF was not performed. Nine of 23 (39%) total patients and 43% of 21 assessable patients had partial responses (PR). The recommended doses for subsequent clinical trials are paclitaxel 225 mg/m2 as a 3-hour infusion followed by carboplatin at a targeted AUC of 7 mg/mL.min. The ability to administer clinically-relevant single agent doses of paclitaxel and carboplatin in combination, as well as the significant antitumor activity noted in this phase I trial, indicate that further evaluations of this regimen in both advanced and early stage NSCLC are warranted. PMID- 9220294 TI - Early clinical investigation of sulofenur with a daily schedule in advanced solid tumours. AB - Sulofenur, a sulfonylurea, has demonstrated antitumour effect in preclinical studies. A phase I trial was initiated to study the clinical aspects. Sulofenur was given p.o. daily for a period of 28 days in 5-week courses. The initial dosage was 250 mg/m2 escalating to 700 mg/m2 daily with no dose modification for the individual patient at any given dose level; 38 patients with advanced solid malignant tumours were enrolled. Haemolytic anaemia was the main side effect. The toxicity was marked at dose levels of 600 and 700 mg/m2. Moderate methaemoglobinaemia also occurred. One case of reversible toxic hepatitis was observed. Generally was ALAT, and more moderately basic phosphatases, and LDH elevated. Tumour regression was not observed but one patient had stable disease throughout nine courses. The maximal detected plasma concentration of Sulofenur in this study was 348 x 10(-6) g/ml. In the present study the maximum tolerated dose (MTD) of Sulofenur was defined to 600 mg/m2. One conclusion from this study is that even at doses above that recommended for future studies-5-600 mg/m2-with this schedule, the suggested effective plasma level from preclinical studies could not be reached. The overall conclusion is that this schedule should not be recommended at all for future studies and the recommendation should be to try to find a schedule in which higher plasma levels can be achieved at a clinically tolerated dose. PMID- 9220293 TI - Phase I trial of fluorouracil modulation by N-phosphonacetyl-L-aspartate and 6 methylmercaptopurine ribonucleoside (MMPR), and leucovorin in patients with advanced cancer. AB - The results of several clinical trials support the hypothesis that biochemical modulation may enhance the antitumor activity of 5-Fluorouracil (5-FU). We have performed a phase I trial using a combination of three different biochemical modulators at the optimal dose established in previous clinical trials. The modulators include: phosphonacetyl-l-aspartate (PALA), which may increase 5-FU incorporation into RNA; leucovorin, which potentiates thymidylate synthase inhibition; and 6-methylmercaptopurine riboside (MMPR), which promotes the intracellular retention of fluorinated nucleotides. The treatment regimen consisted of PALA 250 mg/m2 day 1, followed 24 h later by MMPR 150 mg/m2 as an iv bolus, and the initiation of a 24-hour infusion of 5-FU along with leucovorin 50 mg/m2. This regimen was repeated weekly. Doses of 5-FU were escalated in cohorts of four or more patients from 2,000 to 2,600 mg/m2. Among 20 patients entered, the majority had colorectal cancer, and most had received prior 5-FU treatment. Toxicity was predominantly gastrointestinal, and diarrhea was dose-limiting at a 5-FU dose of 2600 mg/m2. There were three partial remissions observed, two of whom had colorectal cancer. Emerging data that casts doubt on the modulation value of PALA at this dose and schedule suggests that revision of this regimen be considered before Phase II trial. PMID- 9220295 TI - A phase II trial of intravenous vinorelbine in previously untreated patients with extensive small cell lung cancer, a Southwest Oncology Group study. AB - Twenty-two eligible patients with previously untreated extensive small cell lung cancer received intravenous vinorelbine 30 mg/M2 each week until progression. Response was assessed every 4 weeks by chest x-ray or every 8 weeks by CT scan. All responses had to be "confirmed" at all involved sites at least 4 weeks later. Fourteen patients were male and 8 were female with a median age of 64.5 years (range 38-76). Fifteen patients were Caucasian and 7 were African-American. One patient had a "confirmed" partial response, 3 had unconfirmed responses, 13 had stable or progressive disease, and 5 did not have adequate data. The median progression-free survival was 3 months with a median overall survival of 8 months. Thirteen patients experienced 22 episodes of grade 3 toxicity, more than half due to leukopenia and neutropenia, and 1 due to paresthesias. Of 4 episodes of grade 4 toxicity, 1 was due to leukopenia and 3 were due to hyponatremia which was not due to vinorelbine. Significant thrombocytopenia did not occur. The activity of single agent vinorelbine in untreated small cell lung cancer was disappointing when analyzed by Southwest Oncology Group (SWOG) criteria. The median survival in this trial was similar to that found in other SWOG trials using cisplatin based front line therapy and thus confirms previously reported findings that initial treatment with a phase II agent followed by a cisplatin based regimen at progression does not adversely affect overall survival in this population of patients. PMID- 9220296 TI - Phase II trial of oral piritrexim in advanced, previously treated transitional cell cancer of bladder. AB - Oral piritrexim (PTX), a second generation antimetabolite, has been shown to be an active agent against methotrexate refractory transitional cell cancer (TCC) of the bladder in phase I trials. We conducted a phase II trial of this drug in patients with TCC of the bladder who failed a first line chemotherapy regimen. METHODS: Oral PTX was started at the dose of 25 mg three times per day for 5 days weekly for 3 weeks followed by one week of rest. If this was tolerated the dose was increased to 50 mg three times a day. Patients were monitored for response rate and toxicity. RESULTS: Seventeen patients were entered into the trial. Two patients did not complete the required 2 courses of treatment to be evaluable. There were 13 evaluable patients. Among the 13 no one achieved a complete response (CR), however, there were 3 partial responses (PRs = RR: 23%) and 5 stable diseases (SDs). The responses lasted 2, 8 and 14 months. The major dose limiting toxicity was myelosuppression. Two patients died on treatment. One death was due to neutropenic fever and the cause of death in the second patient is thought to be a cerebral vascular accident (CVA). CONCLUSION: PTX is an active drug in the treatment of TCC of the bladder. Bone marrow suppression is the most common dose-limiting toxicity. In view of the observed responses and toxicities in this study and other studies, we suggest that the role of PTX be further investigated in the following clinical settings: 1. Palliative initial treatment in patients with TCC of the bladder who are not candidates for more aggressive chemotherapy. 2. As first line chemotherapy in combination with other active drugs. PMID- 9220298 TI - Predictors of response to cognitive-behavioral group therapy for social phobia. AB - Response to cognitive-behavioral group therapy for social phobia was assessed at posttest and 6-month follow-up in a sample of 62 clients (41 generalized subtype, 21 nongeneralized). Predictors assessed were depression, expectancy, personality disorder traits, clinician-rated breadth and severity of impairment, and frequency of negative thoughts during social interactions. Outcome measures included self-report questionnaires and behavioral tests of dyadic interaction and a public speech. Although no predictor was related to outcome across all domains of measurement, higher depression, more avoidant personality traits, and lower treatment expectancy were each related to poorer treatment response on one or more outcome criteria. Cognitive change was consistently associated with change on self-report symptom measures, but, contrary to expectation, lower rates of negative thinking at posttest did not predict better maintenance of treatment gains at follow-up. PMID- 9220297 TI - An ECOG phase II study of amonafide in unresectable or recurrent carcinoma of the head and neck (PB390). Eastern Cooperative Oncology Group. AB - The purpose of this study was to determine the efficacy and toxicity of amonafide in unresectable or recurrent head and neck cancer and to determine if the degree of toxicity with amonafide correlated with the acetylator phenotype of the patient. Thirty patients were registered on the study and received amonafide, 300 mg/m2, over two hours each day for five consecutive days every 21 days. There was one partial response (3%) which lasted four months. The dose-limiting toxicity was myelosuppression. Acetylator phenotype was determined prior to treatment using HPLC to quantitate caffeine metabolites in urine samples after administration of caffeine. This pharmacokinetic evaluation was performed in 21 patients and revealed that (17/21) 81% of the patients were slow acetylators and 19% of the patients were rapid acetylators. No association was found between acetylator phenotype and toxicity in our patient population. Based on this study, it appears that amonafide given at 300 mg/m2 for 5 consecutive days every 21 days is not active in squamous cell carcinoma of the head and neck, and that acetylator status does not correlate with toxicity. PMID- 9220299 TI - Clinical characteristics of flight phobia. AB - Sixty-six subjects with severe fear of flying were recruited by advertisement and compared to 21 controls without flying fears. Subjects were interviewed and given several questionnaires to determine DSM-III-R diagnoses, history of flying, and development and course of flying phobia. Our phobic sample had a mean age of 46 and was 89% female. Diagnostically, 27% met criteria for current Panic Disorder with Agoraphobia, and 17% criteria for that diagnosis in the past. These two groups were more concerned with internal or social anxiety stimuli during flight than the group who had never had panic attacks but met criteria for Simple Phobia (flying). All three groups were equally concerned about external dangers. Traumatic flight events were common in phobics and controls, but phobics reported reacting to these events more strongly. Our results suggest a vulnerability stress model with several vulnerability factors, including cognitive ones. Treatment implications are discussed. PMID- 9220300 TI - Toward a standard experiment for studying post-treatment return of fear. AB - Fear sometimes returns after successful fear attenuation via in vivo exposure to fear signals. Post-treatment return of fear is of considerable interest both practically and theoretically, but factors associated with return of fear are poorly understood due to conflicting results from procedurally diverse experiments. This paper reports two very similar experiments in which fear of animal specimens was weakened then allowed to return so that factors associated with return of fear could be studied. In each experiment attentional focus versus distraction during exposure served as a between-subjects independent variable. In each case, attempts also were made to predict return of fear via several nonmanipulated variables: initial fear, initial avoidance during voluntary exposure, initial heart rate during voluntary exposure, and speed of fear reduction during repeated exposure trials. With the sample sizes used there was only suggestive evidence that return of fear was associated with distraction during exposure, and with relatively rapid fear decline during exposure. More importantly, the experiments are offered as standard, replicable models for research that will permit procedurally homogeneous investigations of variables with which return of fear is associated. PMID- 9220301 TI - Beliefs about worry and intrusions: the Meta-Cognitions Questionnaire and its correlates. AB - This report describes the development of the Meta-Cognitions Questionnaire to measure beliefs about worry and intrusive thoughts. Factor analyses of the scale demonstrated five empirically distinct and relatively stable dimensions of meta cognition. Four of the factors representing beliefs were: Positive Beliefs About Worry: Negative Beliefs About the Controllability of Thoughts and Corresponding Danger; Cognitive Confidence; and Negative Beliefs about Thoughts in General, including Themes of Superstition, Punishment and Responsibility. The fifth factor represented Meta-Cognitive processes-Cognitive Self-Consciousness-a tendency to be aware of and monitor thinking. The measure showed good psychometric properties on a range of indices of reliability and validity. Scores on the questionnaire subscales predicted measures of worry proneness, proneness to obsessional symptoms, and anxiety. Regression analyses showed that the independent predictors of worry were: Positive Beliefs about Worry; Negative Beliefs About the Controllability of Thoughts and Corresponding Danger: and Cognitive Confidence. Significant differences in particular MCQ subscales were demonstrated between patients with intrusive thoughts, clinical controls and normals. The implications of these findings for models of worry and intrusive thoughts are discussed. PMID- 9220302 TI - Maternal expectations and attributions about coping in anxious children. AB - This study examined maternal expectations and attributions regarding their child's ability to cope with a stressful situation. Children either met DSM III-R criteria for an Anxiety Disorder or were normal. Results indicated that it was not the perception of threat that differentiated the expectations of mothers in both groups, but rather their expectations for coping, both generally and in terms of specific behavior. Mothers of anxiety-disorder (AD) children expected their children to be more upset, less able to make themselves feel comfortable, and were less confident in their children's abilities to perform task related behavior. In general, maternal expectations for coping appear to reflect the actual lower coping ability of anxious children. Concerning attributions, mothers of AD children made fewer causal distinctions between high and low coping than did mothers of normal control (NC) children. Discussion considers how lowered expectations for coping may relate to protective parenting and how such patterns may unwittingly maintain anxious behavior in children. PMID- 9220303 TI - Differentiating post-traumatic stress disorder (PTSD) from major depression (MDD) and generalized anxiety disorder (GAD). AB - Questions about the differential diagnosis of Post-Traumatic Stress Disorder (PTSD) have been raised since this category was reformulated in DSM-III (APA, 1980). Clinicians have reported difficulties distinguishing PTSD from other categories, particularly from Major Depressive and Generalized Anxiety Disorders (MDD and GAD). Diagnostic validity can be established in several ways (e.g., through clinical descriptive studies, laboratory experiments, family history studies, etc.). In this paper, we describe one approach to validation thus far not applied to PTSD: This approach centers directly on whether clinicians can distinguish PTSD from other diagnostic categories. Experienced clinicians were asked to rate the extent to which a common set of 90 symptom items characterized PTSD, MDD, and GAD. Ratings were analyzed with multivariate and univariate analyses of variance and covariance, multiple discriminant function analysis, and factor analysis; moreover, characteristics of rates were examined for possible influences. Results indicated that clinicians readily distinguish PTSD from MDD and GAD as well as MDD from GAD. Findings are presented in terms of univariate analyses, 34 best discriminating items, and factors specifying dimensions differentiating the syndromes of PTSD, MDD, and GAD. Rater characteristics did not influence diagnostic accuracy, although significant differences in magnitude of symptom intensity were found. PMID- 9220304 TI - A comparison of focused and standard cognitive therapy for panic disorder. AB - The relative efficacy of two psychotherapeutic approaches to panic disorder, namely, focused cognitive therapy (FCT) and standard cognitive therapy (SCT) was examined. FCT focused specifically on the "catastrophic misinterpretation" of physical and psychological sensations experienced during panic attacks induced in the office or occurring spontaneously between sessions. SCT focused primarily on the cognitions and beliefs relevant to interpersonal concerns involved in generalized anxiety. We hypothesized that FCT would be more effective than SCT since the latter did not include an induced panic exercise (exposure condition) specific to the patient's panicogenic cognitions. Forty patients diagnosed with panic disorder were randomly assigned to the SCT and FCT groups for approximately 12 to 18 sessions of treatment. Both groups reported significant decreases in the severity of the clinical measures at termination. Moreover, 89.5% of the SCT group and 84.2% of the FCT group were free of panic attacks at 1-year follow-up. Contrary to the predictions, the results for measures of panic attack frequency, anxiety, and depression did not reveal any significant differences between the two groups. Results suggest that in-office "exposure" is not necessary for improvement and that a primary focus on cognitions associated with generalized anxiety may be an effective intervention. However, since improvement in panic was correlated with normalizing of panic-related beliefs in both conditions, it is suggested that cognitive change may be a crucial ingredient of improvement in panic episodes. PMID- 9220305 TI - Jugular venous pressure monitoring: a lost art? PMID- 9220306 TI - Effects of angiotensin II receptor blockade on N-terminal proatrial natriuretic factor plasma levels in chronic heart failure. AB - BACKGROUND: Plasma N-terminal proatrial natriuretic factor (N-terminal proANF) has been shown to reflect intraatrial pressures in heart failure patients, and may be used as a biochemical parameter of atrial pressures. It is still unclear how treatment of heart failure influences the relationship between hemodynamic parameters and plasma levels of N-terminal proANF. METHODS AND RESULTS: This double-blind, placebo-controlled study investigated whether 12 weeks of treatment with the angiotensin II receptor blocker losartan influenced N-terminal proANF plasma levels in 129 chronic heart failure patients. After 12 weeks of treatment, N-terminal proANF level was increased in patients given placebo by 344 +/- 1126 pmol/L, whereas in patients given 50 mg losartan, the levels had decreased by 251 +/- 886 pmol/L (P < .01 vs placebo). The change in N-terminal proANF correlated significantly with the corresponding change in pulmonary capillary wedge pressure (r = .53, P < .001). CONCLUSION: The correlation between N-terminal proANF plasma levels and pulmonary capillary wedge pressure at baseline, as well as the correlation between the changes in these parameters during treatment, suggests that plasma N-terminal proANF may be used to monitor effects on left ventricular filling pressures in patients treated with losartan. PMID- 9220307 TI - Plasma N-terminal atrial natriuretic factor: a predictor of survival in patients with congestive heart failure. AB - BACKGROUND: Congestive heart failure results in biatrial stretch, which stimulates myocyte release of atrial natriuretic factor (1-126). The N-terminal fragment, proatrial natriuretic factor(1-98), (proANF), is released on an equimolar basis with the C-terminal (99-126) active hormone and may be assayed simply because of prolonged in vitro stability. Proatrial natriuretic factor has been shown to be predictive of clinical status in patients with congestive heart failure. This retrospective analysis was undertaken to evaluate the relationship between N-terminal atrial natriuretic factor(1-98) and survival in patients with stable congestive heart failure. METHODS AND RESULTS: Proatrial natriuretic factor was sampled from 316 patients (mean age, 68 (+/-) 11 years; 71% men) recruited from an outpatient heart failure clinic. The mean ejection fraction was 34 (+/-) 13%. Seventy-three deaths were registered during the period of data collection (42 months). Deaths per proANF quartile (n = 79) were as follows: 2 (2.5%) in quartile I. 13 (16.5%) in quartile II, 21 (26.6%) in quartile III, and 37 (46.8%) in quartile IV. The odds ratio estimates for death adjusted for age and sex were 7.6, 13.9, and 33.9 for the second, third, and fourth quartiles, respectively. Survival curves constructed according to proANF quartiles demonstrate significant differences in mortality rates. The correlation with death was greater for proANF as compared with left ventricular end-diastolic diameter (P < .001), systolic pulmonary artery pressure (P < .005), or ejection fraction (P < .05). CONCLUSION: These data indicate that the concentration of proANF is related to prognosis in patients with heart failure and that moderate elevation is associated with markedly decreased survival. Analysis should be of practical value in the assessment of prognosis in this heterogeneous population. PMID- 9220308 TI - Vasodilation by urapidil in the treatment of chronic congestive heart failure in addition to angiotensin-converting enzyme inhibitors is not beneficial: results of a placebo-controlled, double-blind study. AB - BACKGROUND: The therapeutic benefit of an angiotensin-converting enzyme (ACE) inhibitor in combination with a different type of vasodilator is unknown. METHODS AND RESULTS: To evaluate the effects of a combined therapy on quality of life, exercise tolerance, and hemodynamic parameters, patients with severe heart failure (New York Heart Association classes III and IV, ejection fraction below 35%) who were on ACE inhibitor therapy were randomly assigned to additional double-blind treatment with urapidil (60-120 mg/d) or placebo for 12 weeks. After enrollment of 36 patients, the study was terminated early because no beneficial effects on exercise tolerance and hemodynamic parameters could be shown for the urapidil treatment, and a trend toward increased mortality of the urapidil group was observed (odds ratio, 4.92 [0.49-49.6]; P = .167). CONCLUSION: The combination of urapidil with an ACE inhibitor in the treatment of severe chronic congestive heart failure does not seem to offer any advantages over therapy with an ACE inhibitor alone and may have potentially harmful effects. PMID- 9220309 TI - Human leukocyte antigen class II associations in patients with idiopathic dilated cardiomyopathy. Myocarditis Treatment Trial Investigators. AB - BACKGROUND: Idiopathic dilated cardiomyopathy (IDC) is a disease of unknown etiology for which immune abnormalities, possibly related to viral infections, are suspected but unproven. Previous serologic studies have reported associations between human leukocyte antigen DR4 and IDC. A molecular study of human leukocyte antigen associations was undertaken in patients with IDC to further explore the possibility of susceptibility markers of genetically determined disease. METHODS AND RESULTS: In this study, 36 patients from the Myocarditis Treatment Trial (32 IDC and 4 myocarditis patients) were examined using restriction fragment length polymorphism analysis and polymerase chain reaction amplification with sequence specific primers to perform class II typing. All 4 myocarditis patients were DQ5 positive and 3 possessed the allele DQB1*0501. In the IDC group, the frequency of human leukocyte antigen DR4 was similar to that reported in the normal population. In addition, there was no excess prevalence of any molecularly defined DR4 alleles (0401-0419). There was an increase in the frequency of DR12 in IDC patients. The frequencies of the alleles DQB1 *0503 and DQB1*0301 and/or *0304 were also increased in IDC patients versus the normal population. CONCLUSION: The molecular studies point to a relationship between the DQ locus and IDC. PMID- 9220310 TI - Removal from the "waiting list" for heart transplantation: a risk factor for sudden death? AB - METHODS AND RESULTS: Over an 18-month period, the patients on the heart transplantation waiting list at our institution were evaluated to determine if continued listing was appropriate. Ten patients were removed because of significant improvement in clinical status and exercise capacity (n = 9) or because of criteria violation (n = 1). Four of these patients died suddenly and unexpectedly within 4 months of delisting, resulting in a 6-month survival of 60% for the patients removed. During the same period, the 6-month survival for newly listed patients (n = 10) was 80% and that for newly transplanted patients (n = 13) was 92%. An elevated pulmonary capillary wedge pressure (> or = 18 mmHg) was the only clinical or laboratory feature that appeared to distinguish the four patients who died suddenly following delisting. CONCLUSION: The results of this preliminary study suggest that removal of a patient from a heart transplant waiting list may represent a risk for sudden death, particularly in patients with elevated ventricular filling pressures, irrespective of otherwise favorable clinical status and exercise performance. PMID- 9220311 TI - Cardiac-specific overexpression of tumor necrosis factor-alpha causes lethal myocarditis in transgenic mice. AB - BACKGROUND: Tumor necrosis factor (TNF)-alpha, a proinflammatory cytokine with negative inotropic effects, can be detected in myocardium with end-stage heart failure, after endotoxin administration, and during transplant rejection. Various studies suggest that TNF-alpha participates in the pathogenesis of cardiac dysfunction. To test this hypothesis, transgenic mice were made that selectively overexpress TNF-alpha in cardiomyocytes. METHODS AND RESULTS: A transgene construct was made containing the murine alpha-myosin heavy chain promoter and the coding sequence of murine TNF-alpha, followed by the simian virus 40 T antigen intron and polyadenylation signals. Injection of this construct into fertilized eggs yielded three transgenic mice, all of which died spontaneously before the completion of weaning. Gross pathologic analysis of these mice demonstrated a decrease in body weight with markedly increased heart weight. Histologic examination of the heart revealed a substantial, diffuse lymphohistiocytic inflammatory infiltrate, associated with interstitial edema. Reverse transcriptase polymerase chain reaction showed that the transgene was expressed in the heart. Enzyme-linked immunosorbent assay demonstrated a substantial amount of TNF-alpha protein in the transgenic heart. CONCLUSION: Overexpression of TNF-alpha in the heart leads to severe myocarditis and cardiomegaly. These results support the hypothesis that myocardial expression of TNF-alpha can contribute to the pathogenesis of cardiac dysfunction. PMID- 9220312 TI - Early pharmacologic intervention may prevent the deterioration in endothelial function after experimental myocardial infarction in rats: effects of ibopamine and captopril. AB - BACKGROUND: Endothelial function is progressively disturbed after myocardial infarction (MI), which may be related to both neurohumoral activation and hemodynamic alterations. Consequently, it may be suggested that drugs that favorably affect these factors may also have a positive effect on endothelial function. METHODS AND RESULTS: Rats underwent coronary ligation (n = 24) or remained unoperated (n = 21), and were randomized to captopril (25 mg/kg/d) or ibopamine (10 mg/kg/d) or remained untreated. Treatment was started following MI and lasted 8 weeks, after which rats were sacrificed for in vitro studies. Left ventricular end-systolic pressure was higher in rats treated with captopril (83 +/- 6 mmHg) and ibopamine (80 +/- 3 mmHg), as compared with untreated MI rats (48 +/- 6 mmHg, P < .01 for both). Increased plasma norepinephrine levels in MI rats were reduced by captopril and ibopamine (both P < .05). Infarct size was smaller in rats treated with captopril (26.7 +/- 3.6%, P < .05) and ibopamine (31.4 +/- 4.3%, P = NS), as compared with untreated rats (41.7 +/- 2.4%). Maximal endothelium-dependent relaxation (Emax; % precontraction) and the concentration of methacholine causing 50% Emax, expressed as negative log(pIC50) were significantly reduced in aortic rings from MI control subjects (pIC50 = 6.15 +/- 0.06 mol/L, Emax = 32.0 +/- 4.2%), as compared with normal control subjects (pIC50 = 6.57 +/- 0.07 mol/L, P < .001; Emax = 50.0 +/- 4.9%, P = .022). Captopril (pIC50 = 6.30 +/- 0.08 mol/L, Emax = 45.1 +/- 7.0%) and ibopamine (pIC50 = 6.60 +/- 0.08 mol/L, Emax = 43.8 +/- 5.2%) improved these parameters in MI rats. CONCLUSION: The results demonstrate preservation of endothelial function by early pharmacologic intervention after experimental MI in rats in the setting of concomitant reduction in infarct size. PMID- 9220313 TI - Antihypertensive and cardioprotective effects after angiotensin-converting enzyme inhibition: role of kinins. AB - Kinins are potent bioactive peptides formed by the enzymatic action of kallikrein on kininogens. The discovery that angiotensin-converting enzyme, which generates angiotensin II, is also a major degrading enzyme of kinins, gave rise to the hypothesis that kinin potentiation, in addition to angiotensin II reduction, may be involved in the therapeutic actions of angiotensin-converting enzyme inhibitors. Angiotensin-converting enzyme inhibitors have become important drugs in the treatment of hypertension, congestive heart failure, postmyocardial infarction, and diabetic nephropathy. Although angiotensin II reduction appears to be the predominant mechanism of the antihypertensive effect of chronic angiotensin-converting enzyme inhibitor treatment, the role of kinins in the antihypertensive effects of angiotensin-converting enzyme inhibitors seems to be renin dependent and cannot be generalized for all models of hypertension. On the other hand, at least under experimental conditions, various cardioprotective effects of angiotensin-converting enzyme inhibitors appear to be due to the potentiation of endogenous kinins, including improved cardiac function, structural changes following myocardial ischemia, and induction of capillary growth in hypertension-induced left ventricular hypertrophy. PMID- 9220314 TI - Identification of a T-cell receptor from a therapeutic murine T-cell clone. AB - Tumor-infiltrating lymphocytes (TIL) have been successfully used for the treatment of metastatic malignancies in clinical trials and in experimental animal models. Tumor-specific reactivity by TIL is mediated via receptors expressed on the surface of T cells (TcRs), which recognize tumor-associated antigens (TAA) presented in the context of MHC molecules on the surface of tumor cells. The current study was performed to identify the TcR alpha and beta chains from a tumor-specific therapeutic TIL clone that can be used to develop a preclinical animal model for genetically modifying lymphocytes and hematopoietic progenitors with TcR genes. TIL 205 was generated from a subcutaneous implant of MCA-205 fibrosarcoma and at 21 days was cloned by limiting dilution. TIL clone 8, obtained from a culture seeded at one cell/well, mediated specific lysis and specific secretion of gamma-interferon to MCA-205 and WP6, a subclone of MCA 205. No reactivity was observed against other syngeneic sarcoma lines. Anchor polymerase chain reaction analysis determined that antigen recognition by clone 8 was mediated by a TcR consisting of V alpha 3/J alpha 27 and V beta 8.2/D beta 2.1/D beta 2.4. Immunofluorescent staining with V beta subfamily specific monoclonal antibodies revealed that > 95% of the T cells in TIL clone 8 expressed V beta 8.2, confirming that TIL clone 8 was indeed a clone. In contrast, approximately 30% of the T cells in the parental TIL 205 expressed V beta 8.2. The transfer of as few as 500,000 TIL clone 8 cells in conjunction with the systemic administration of recombinant human interleukin-2 mediated regression of established 3-day WP6 lung metastases. Thus, clone 8 recognizes a biologically relevant tumor rejection antigen, making the V alpha 3/J alpha 27-V beta 8.2/D beta 2.1/J beta 2.4 TcR isolated from this clone useful as a probe for cloning the tumor-rejection antigen in the WP6 tumor as well as modeling, in mice, the TcR-based gene therapies being developed for humans. PMID- 9220315 TI - B7-1(CD80)-transfected human glioma cells and interleukin-12 directly stimulate allogeneic CD8+ T cells. AB - To obtain more effective cytotoxic T lymphocytes (CTLs), we examined the effect of B7 costimulation and interleukin-12 (IL-12) on the induction of allogeneic CTLs. Peripheral blood lymphocytes (PBLs) or positively selected human CD8+ T cells (purity > 99%) from healthy donors were used as effector cells, and B7-1 transfected or nontransfected U251 human glioma cells (U251-B7 or U251-Vec.) were used as stimulator cells. In mixed lymphocyte culture (MLC) with PBLs and U251 cells, nonspecific natural killer (NK) activity was raised by addition of IL-12, and the effect of B7 costimulation was not observed. However, in MLC with CD8+ T cells, efficient proliferation, generation of CTLs, and cytokine production were induced by MLC with U251-B7 in the presence of IL-12. Efficient generation of CTLs was not induced by either MLC with U251-B7 alone or the addition of IL-12 alone. Our results indicate that a combination of B7-1 costimulation and IL-12 is effective for inducing generation of CTLs, and that the CD8+ T cells can be differentiated into CTLs without the help of CD4+ T cells or antigen-presenting cells or both. PMID- 9220316 TI - Expression of MAGE genes in ocular melanoma cell lines. AB - The pathobiology of melanomas that develop within the eye is distinct from melanomas that develop within the subcutaneous tissues of the skin. This may be related to the unique structural and functional differences between normal melanocytes present within the uveal tract of the eye and the epidermal layers of the skin. The purpose of the present study was to determine whether normal pigmented cells within the eye (melanocytes and retinal pigment epithelial cells) and cultured cells derived from malignant ocular melanomas express the MAGE genes that encode tumor antigens that are recognized by specific CD8+ cytotoxic T lymphocytes. In the present series of experiments, we examined MAGE expression in cultured ocular melanoma cells obtained from a group of 17 ocular melanoma patients. Normal ocular melanocytes and retinal pigment epithelial cells were recovered and cultured from eyes enucleated for trauma. MAGE gene expression was determined using reverse transcription-polymerase chain reaction specific for either MAGE-1, -2, or -3. Our results demonstrate that MAGE-1, MAGE-2, and MAGE-3 genes are transcribed in primary ocular melanoma cell lines and are detected in cells recovered from 41, 53, and 53% of the patients examined, respectively. Normal choroidal melanocytes and retinal pigment epithelial cells did not express MAGE genes. We conclude that cultured ocular melanoma cell lines express MAGE-1, MAGE-2, and MAGE-3 genes. PMID- 9220318 TI - Prophylactic intervention in RadLV-induced lymphomagenesis with an anti-IL-4 monoclonal antibody. AB - Intrathymic inoculation of the radiation leukemia virus (RadLV) into C57BL/6 mice induces thymic lymphomas after 4-6 months. During premalignant latency, a population of RadLV-infected prelymphoma (PL) cells (whose survival is dependent on autostimulation with IL-4) persists in the thymus. PL cells explanted from RadLV-inoculated mice can be propagated in cultures containing IL-4, and in vitro growth of PL cells is effectively inhibited by anti-IL-4 antibodies. We subjected RadLV-inoculated mice to prophylactic treatment with anti-IL-4 antibodies and a virus-specific immunotoxin (IT). Administration of IT delayed the onset of lymphoma but was not curative. Anti-IL-4 antibodies had a similar effect when administered at low doses. High doses of anti-IL-4, given 3-5 weeks after virus inoculation, provided complete protection against lymphoma. These results demonstrate the effectiveness of prophylactic intervention during premalignancy by using antagonists that restrain the growth of PL cells. PMID- 9220319 TI - A prospective randomized evaluation of the prophylactic use of low-dose dopamine in cancer patients receiving interleukin-2. AB - The administration of high-dose interleukin-2 (IL-2) causes tumor regression in 17-25% of patients with metastatic melanoma or renal cell carcinoma. Renal dysfunction is a common dose-limiting toxicity of IL-2 administration, limiting 26% of treatment cycles. We have conducted a prospective randomized trial to evaluate whether the prophylactic administration of low-dose dopamine (2 mg/kg/min) can minimize renal toxicity and thus affect the amount of IL-2 administered. Forty-two patients were randomly assigned to receive systemic high dose IL-2 with standard supportive measures (group A = 21 patients) or with the addition of prophylactic dopamine (group B = 21 patients) at 2 mg/kg/min. For patients in group B, dopamine was instituted 1 h before the initiation of IL-2 administration and was discontinued 6-12 h after the maximum number of doses of IL-2 were given. There was no difference in the amount of IL-2 administered for each course of therapy for groups A and B. Despite differences in urine flow (milliliters per kilogram per day), fluid balance (liters per day), and overall weight gain, prophylactic low-dose dopamine did not significantly alter maximum plasma urea or creatinine levels in group B when compared with the control group (group A). The overall toxicity profile considering all grade 3 and 4 toxicities for patients in groups A and B was comparable. Thus, there is no evidence to support the routine use of prophylactic low-dose dopamine in patients receiving high-dose IL-2. PMID- 9220317 TI - Dendritic cells infected with poxviruses encoding MART-1/Melan A sensitize T lymphocytes in vitro. AB - Dendritic cells (DC) are potent professional antigen-presenting cells that can activate naive T lymphocytes and initiate cellular immune responses. As adjuvants, DC may be useful in enhancing the immunogenicity of tumor antigens and mediating tumor regression. Endogenous expression of antigen by DC offers the potential advantage of allowing prolonged constitutive presentation of endogenously processed epitopes and exploitation of multiple restriction elements for the presentation of the same antigen. In this report, we show that human DC are (a) capable of infection by recombinant poxviruses encoding melanoma associated antigen (MAA) genes and (b) capable of efficiently processing and presenting these MAA to cytotoxic T cells. In 6/6 HLA A*0201-expressing melanoma patients tested, the virally driven expression of MART-1/Melan A MAA by DC was sufficient to generate CD8+ T lymphocytes that could recognize naturally processed epitopes on tumor cells. In most cases, specific anti-MART-1 reactivity could be detected after a single stimulation. Analysis of epitope dominance revealed that the amino acid sequence recognized by these cytotoxic T lymphocytes (CTL) corresponded to the MART-1(27-35) residues previously shown to be most commonly recognized by cytotoxic T lymphocytes expanded from metastatic melanoma lesions. These data show that the virally driven expression of MAA by DC can be exploited for the efficient induction of clinically relevant cytotoxic T-cell responses. This has clinical implications for active immunization therapy, and currently vaccine trials have been proposed for patients with metastatic melanoma. PMID- 9220320 TI - Phase II trial of interleukin-2 and interferon-alpha in patients with renal cell carcinoma: clinical results and immunologic correlates of response. AB - A phase II trial was conducted in patients with metastatic renal cell carcinoma, to assess the clinical efficacy and immunoregulatory effects of continuous infusion recombinant interleukin-2 (rIL-2) (9.0 x 10(6) IU/m2/day on days 1-5, 8 12, 15-19, and 22-26) and subcutaneously administered recombinant human interferon-alpha 2b (rHuIFN alpha 2b) (10.0 x 10(6) U/m2/day TIW). Thirty-six patients with metastatic renal cell carcinoma, performance status of 0-1, and measurable disease who had not received prior rIL-2, rHuIFN alpha 2b, or chemotherapy were treated. Patients with CNS metastases, active infections, history of another malignancy within 3 years, and those requiring corticosteroids were ineligible. Cycles of rIL-2 and rHuIFN alpha 2b were administered in the outpatient department every 6-8 weeks in stable or responding patients until patient tolerance or a complete response were reached. Doses were modified for grade III or IV toxicity. Ancillary studies included three-color immunocytometric analysis of peripheral blood lymphocytes, repetitive tumor biopsies for immunohistologic analysis of infiltrating cells and proliferative responses of tumor infiltrating lymphocytes, and preliminary studies of changes in peripheral blood T-lymphocyte signal transduction molecules [T-cell receptor (TCR)-zeta, p56ick, p59fyn]. Thirty-six eligible patients were treated, with 6 of 36 patients (17%, 95% confidence interval 6-33%) responding (3 complete response, 3 partial response). In two of the partial responders, and in an additional three patients with either minimal tumor regression (one patient) or stable disease (two patients), surgical removal of residual disease was undertaken. The median survival of all patients was 14 months. The toxicity of this regimen was severe, but outpatient administration was possible in most instances. Immunoregulatory effects on T-cell subsets included increases in various CD3+ CD25+/- HLADr+/- subsets unrelated to response. Tumor biopsies before and/or during therapy were obtained in 17 patients, and no consistent alterations in the degree of T lymphocyte or macrophage infiltrates could be detected. In a subset of patients, tumor infiltrating lymphocyte proliferative responses and levels of peripheral blood T-cell signal transduction molecules (TCR-zeta, p56lck, p59fyn) were investigated. Abnormalities were found in selected patients, which improved during rIL-2/rHuIFN alpha 2b therapy. This cytokine combination produces tumor regression in selected patients with metastatic renal cell carcinoma. Surrogate immunologic markers associated with response were not identified; however, preliminary studies demonstrate investigation of immune defects and their reversal with cytokine therapy is possible. PMID- 9220321 TI - High-dose regimen of interleukin-2 and interferon-alpha in combination with lymphokine-activated killer cells in patients with metastatic renal cell cancer. AB - Seventy-two patients with metastatic renal cell cancer were treated with the combination of high-dose interleukin-2 (IL2), interferon-alpha (IFN alpha), and lymphokine-activated killer cells (LAK). Seventeen patients were entered in a feasibility part of the study (protocol 1) and 55 in an efficacy part (protocol 2). Protocol 2 differed from protocol 1 in the addition of IFN alpha to the first 5 days of IL2 infusion. Each patient was planned to receive two induction cycles. IL2, 18 MIU/m2/day, was administered continuously i.v. on days 1-5, and IFN alpha, 5 MIU/m2/day (protocol 2), was administered i.m. on days 1-5, followed by three daily lymphaphereses on days 7-9. On day 12, treatment was resumed with IL2 and IFN alpha on days 12-15 and LAK reinfusions on days 12-14. In protocol 1, three complete (CR) and one partial (PR) responses were achieved (response rate 24%). The median duration of response and the median survival were 18.1 and 13.9 months, respectively. The 3-year survival was 35%. Of the 51 evaluable patients in protocol 2, 6 achieved a CR and 13 a PR (response rate 37%). The median duration of response was 11.1 months. The median survival was 16.9 months. The 3 year survival was 35%. There were three treatment-related deaths. Other severe toxicities included hypotension, cardiotoxicity, pulmonary edema, renal toxicity, and infectious complications. In the two induction cycles, only 54 and 42% of the planned doses could be administered. We conclude that the use of high-dose regimens of IL2 and IFN alpha is not warranted, unless we can define more accurately which patients may experience long-term survival as a result of treatment. PMID- 9220322 TI - Compensating subjects of medical research. PMID- 9220323 TI - Petty corruption in health care. PMID- 9220324 TI - Genetic screening: a comparative analysis of three recent reports. AB - Three recent reports on genetic screening published in the United Kingdom, Denmark and the Netherlands are discussed. Comparison of the Dutch report with the Danish and the Nuffield reports reveals that the Dutch report focuses on the aim of enlarging the scope for action, emphasising protection of autonomy and self-determination of the screenee more than the other two reports. The three reports have in common that the main concern is with concrete issue such as stigmatisation, discrimination, protection of the private sphere and issues linked with labour and insurance. Some potential long term consequences, however, tend to be neglected or underestimated. These omissions are pointed out. PMID- 9220325 TI - Requests for "inappropriate" treatment based on religious beliefs. AB - Requests by patients or their families for treatment which the patient's physician considers to be "inappropriate" are becoming more frequent than refusals of treatment which the physician considers appropriate. Such requests are often based on the patient's religious beliefs about the attributes of God (sovereignty, omnipotence), the attributes of persons (sanctity of life), or the individual's personal relationship with God (communication, commands, etc). We present four such cases and discuss some of the basic religious tenets of the three Abrahamic faith traditions as they relate to such requests. We suggest that religious reasons for requesting "inappropriate" treatment are "special" and deserve serious consideration. We offer guidance to assist clinicians and clinical ethicists as they attempt to resolve these conflicts, emphasising the importance of understanding the religious beliefs of the patient/surrogate and suggesting the assistance of a religious interpreter. We suggest open discussion with patients and families of both the clinical situation and the theological basis for these requests. We also suggest that clinicians use additional religious doctrines or principles from patients' own traditions to balance the reasons behind the requests. We conclude that most persistent requests for "inappropriate" treatment should be honoured. PMID- 9220326 TI - Medical futility and the social context. AB - The concept of medical futility has come to be seen in some quarters as a value neutral trump card when dealing with issues of power and conflicting values in medicine. I argue that this concept is potentially useful, but only in a social context that provides a normative framework for its use. This social context needs to include a broad consensus about the purpose of medicine and the nature of the physician-patient relationship. PMID- 9220328 TI - Commentary. 1: The right to refuse treatment. PMID- 9220327 TI - The right to refuse treatment is not a right to be killed. AB - It is widely accepted now that a patient's right to refuse treatment extends to circumstances in which the exercise of that right may lead to the patient's death. However, it is also often effectively assumed, without argument, that this implies a patient's right to request another agent to intervene so as to bring about his or her death, in a way which would render that agent guilty of murder in the absence of such a request. But the right to refuse treatment can, logically, have no such implication, and the mistaken supposition that it does conflates a right to die with a right to be killed. Confusion over this issue is brought out by an examination of conflicting opinion concerning the permissible termination of ventilation for mentally competent patients. A wider lesson may be drawn regarding the need for the ethical assessment of new forms of life sustaining medical technology. PMID- 9220329 TI - Commentary. 2: Thesis correct: argument unconvincing. PMID- 9220330 TI - Decision-making in the critically ill neonate: cultural background v individual life experiences. AB - OBJECTIVES: In treating critically ill neonates, situations occasionally arise in which aggressive medical treatment prolongs the inevitable death rather than prolonging life. Decisions as to limitation of neonatal medical intervention remain controversial and the primary responsibility of the generally unprepared family. This research was designed to study response patterns of expectant mothers towards treatment of critically ill and/or malformed infants. DESIGN/SETTING: Attitudes were studied via comprehensive questionnaires divided into three sections: 1-Sociodemographic data and prior personal experience with perinatal problems; 2-Theoretical philosophical principles used in making medical ethical decisions; and 3-Hypothetical case scenarios with choices of treatment options. SUBJECTS AND RESULTS: Six hundred and fifty pregnant women were studied. Maternal birthplace (p = 0.005) and level of religious observance (p = 0.02) were strongly associated with the desire for maximally aggressive medical intervention in the hypothetical case scenario. Specific personal experiences such as infertility problems, previous children with serious mental or physical problems were not correlated with the selection of different treatment choices. Of the theoretical principles studied, only the desire to preserve life at all costs was significantly associated with the choice for maximal medical treatment (p = 0.003). CONCLUSIONS: Maternal ethnocultural background and philosophical principles more profoundly influenced medical ethical decision-making than did specific personal life experiences. PMID- 9220331 TI - Organ transplant initiatives: the twilight zone. AB - Assessments of the acceptability of new transplantation practices require a pinpointing of not only the meaning of death, but also the timing of death. They typically perceive elective ventilation as occurring just prior to death and non heart-beating donor protocols as operative just after death. However, such practices in fact highlight the general vagueness and ambiguity surrounding these issues in both law and ethics. Supply-side dilemmas in transplantation lend real urgency to this "life or death" debate. PMID- 9220332 TI - Bodies, rights and abortion. AB - The issue of abortion is discussed with reference to the claim that people have a right of control over their own bodies. Do people "own" their own bodies? If so, what would be entailed? These questions are discussed in commonsense terms and also in relation to the jurisprudence of Hohfeld, Honore, Munzer and Waldron. It is argued that whether or not women are morally and/or should be legally entitled to have abortions, such entitlements cannot be derived from a general moral entitlement to do what we will with our own bodies since there is no such entitlement. Whether or not we "own" them, we can have rights duties, liabilities, restrictions and disadvantages as well as rights concerning our own bodies. PMID- 9220333 TI - Compensation for subjects of medical research: the moral rights of patients and the power of research ethics committees. AB - Awareness of the morally significant distinction between research and innovative therapy reveals serious gaps in the legal provision for compensation in the UK for injured subjects of medical research. Major problems are limitations inherent in negligence actions and a culture that emphasises indemnifying researchers before compensating victims. Medical research morally requires compensation on a no-fault basis even where there is proper consent on the part of the research subject. In particular, for drug research, there is insufficient provision in the current patient guidelines of the Association of the British Pharmaceutical Industry, since they make "no legal commitment" to paying compensation for injury to patient subjects. There is a need for the provision of both adequate insurance and contractual arrangements for making payments. The solution is for Local Research Ethics Committees (LRECs) to make use of their power to withhold approval of medical research where compensation is not legally enforceable. PMID- 9220334 TI - The views of members of Local Research Ethics Committees, researchers and members of the public towards the roles and functions of LRECs. AB - BACKGROUND: It can be argued that the ethical conduct of research involves achieving a balance between the rights and needs of three parties-potential research participants, society, and researchers. Local Research Ethics Committees (LRECs) have a number of roles and functions in the research enterprise, but there have been some indications that LREC members, researchers and the public can have different views about these responsibilities. Any such differences are potential sources of disagreement and misunderstanding. OBJECTIVES: To compare the views of LREC members, researchers and the public towards the roles and functions of LRECs. DESIGN: A questionnaire that contained items concerned with a variety of such roles was distributed to general practice patients (as proxies for potential research participants), researchers and LREC members. FINDINGS: While general practice patients believed that the main function of LRECs is to ensure that research participants come to no harm, LREC members were more concerned with the protection of participants' rights. There was also some disagreement between members and researchers with regard to the consideration of proposals on the grounds of scientific merit. CONCLUSIONS: Local Research Ethics Committee members need to be aware of potential differences in views, that they ought to make their priorities clear, and that membership of LRECs ought to reflect the views of both researchers and potential research participants. PMID- 9220335 TI - Workplace urine screening for drug abuse. PMID- 9220336 TI - Informed consent in Turkey. PMID- 9220337 TI - Why are ATP depletion rates in situ in ischemic myocardium so much lower than one might predict from the activity of the mitochondrial ATPase in sonicated heart mitochondria? PMID- 9220338 TI - Stretch-activated ion channels in the heart. PMID- 9220339 TI - Retinoic acid accelerates embryonic stem cell-derived cardiac differentiation and enhances development of ventricular cardiomyocytes. AB - Pluripotent embryonic stem (ES) cells spontaneously differentiate via embryo-like aggregates into cardiomyocytes of pacemaker-, atrium- and ventricle-like type, which can be distinguished by their specific patterns of action potentials. It has been shown that retinoic acid (RA) treatment during ES cell differentiation increases the number of cardiomyocytes in a time- and concentration-dependent manner. In order to test the effect of RA on cardiomyocyte differentiation and specialization into ventricle-like cardiomyocytes, we studied gene expression of beta-galactosidase driven by the ventricular myosin light chain-2 (MLC-2v) promoter as an indicator for ventricular differentiation. Clones containing the stably integrated expression vector pGNA/MLC-2.1 were selected, which revealed an increase of beta-galactosidase activity in cardiomyocytes of embryoid bodies at day 7 + 16. RA, both, in the all-trans and in the 9-cis configuration resulted in a significant acceleration of cardiomyocyte differentiation and a transient increase of beta-galactosidase activity. To test whether this acceleration of cardiac differentiation and RA-induced increase of the MLC-2v promotor/beta galactosidase activity reflects an increase of cardiac- and ventricle-specific gene expression, a semi-quantitative RT-PCR analysis was performed for alpha cardiac myosin heavy chain (alpha-MHC) and MLC-2v genes. It was shown that both 10(-8) M and 10(-9) M RA resulted in an increased level of alpha-cardiac MHC and MLC-2v mRNA in embryoid bodies in early, but not in terminal developmental stages. This led us to the conclusion that the RA-induced accelerated expression of cardiac-specific genes results in an enhanced development of ventricular cardiomyocytes. An increased number of ventricle-like cells after RA treatment was also found by patch-clamp analysis. The number of cardiomyocytes with Purkinje- and ventricle-like properties was shown to be increased by RA, whereas the number of pacemaker- and atrium-like cells was reduced and early pacemaker cells were not quantitatively affected. PMID- 9220340 TI - Formation of binucleated cardiac myocytes in rat heart: I. Role of actin-myosin contractile ring. AB - Cardiac myocytes in rat hearts lose their ability to undergo cytokinesis between day 3 and day 4, resulting in the formation of binucleated myocytes. Failure in the formation of the actin-myosin contractile ring could cause cardiac myocytes to be defective in cytokinesis. Enzymatically isolated cardiac myocytes from 2- and 4-day-old rats were employed to investigate the organisation and distribution of actin, myomesin, and myosin by rhodamine phalloidin, anti-myomesin, and isoform-specific anti-myosin antibodies, respectively. Interestingly, the actin myosin contractile ring was formed in mitotic myocytes from both 2- and 4-day-old animals. The changes in organisation and distribution of actin, myosin and myomesin in mitotic myocytes from 4-day-old rats were similar to those from 2-day old rats, except that there were longitudinal actin filaments in the cytoplasm of mitotic myocytes from 4-day-old rats. In mitotic myocytes from both 2- and 4-day old rats, actin disassembled in prometaphase, concentrated in the equator of the mitotic spindle in late anaphase, and formed a circumferential intensely staining band in early telophase. Cytoplasmic myosin was evenly distributed in the cytoplasm as small spots, and appeared to associate with the cell membrane from interphase to early anaphase. It became progressively more concentrated in association with the cortical membrane in the equator region in late anaphase, formed a ring-like structure in early telophase, and remained associated with adjacent membrane at the cleavage furrow until late telophase. Sarcomeric myosin and myomesin were only partially disassembled in mitotic myocytes from both 2- and 4-day-old animals. The present study showed that the actin-myosin contractile ring was actually formed during the binucleation process of cardiac myocytes. Molecules involved in the latter stages of cytokinesis may be responsible for incomplete cytokinesis during the binucleation process. PMID- 9220341 TI - Formation of binucleated cardiac myocytes in rat heart: II. Cytoskeletal organisation. AB - Neonatal cardiac myocytes continue to undergo nuclear division, but lose their ability to complete cell division between 3 and 4 days of age. To examine cytoskeletal organisation of cardiac myocytes during mitosis, freshly isolated cardiac myocytes from 2-, 4-, 6- and 8-day-old rats were fixed and labeled with anti-tubulin, vinculin, desmin and sarcomeric alpha-actinin antibodies. The central, nuclear region of cardiac myocytes is expanded to form a balloon-like structure when they entered prophase. The organisation of microtubules, vinculin and desmin in mitotic myocytes from 4-, 6- and 8-day-old rats was identical to that in dividing myocytes from 2-day-old animals. Microtubules emanating from the nuclear membrane mainly ran along the longitudinal axis of cardiac myocytes in interphase. Microtubules were disassembled and reorganised into the mitotic spindle during mitosis. Desmin was disassembled, either diffusely distributed in the cytoplasm or formed spotty cytoplasmic aggregates during mitosis. Vinculin was disassembled in prometaphase, diffusely distributed in the cytoplasm and associated with cell membranes. During telophase it concentrated in the equator of mitotic spindles. Sarcomeric alpha-actinin became dispersed in the cytoplasm of mitotic myocytes from 2-day-old rats in prometaphase. It remained diffusely distributed in the cytoplasm and associated with cell membranes until the completion of cytokinesis. However, sarcomeric alpha-actinin was only partially disassembled in 4-, 6- and 8-day-old myocytes. Striations of alpha-actinin with full sarcomere length were observed in the cytoplasm as well as in the region of furrow formation. Thus, incomplete disassembly and presence of myofibrils in the equator region where cleavage furrows from may physically impede the furrowing of sarcolemma driven by the contractile ring, resulting in the formation of binucleated cardiac myocytes. PMID- 9220342 TI - Role of inducible nitric oxide synthase in pharmacological "preconditioning" with monophosphoryl lipid A. AB - Pretreatment with monophosphoryl lipid A (MLA) can pharmacologically mimic the second window of ischemic preconditioning (SWOP) to protect the heart from prolonged ischemia and reperfusion injury. Based on the delayed time course for development of MLA associated cardioprotection, this study was designed to test if MLA's cardioprotective effect is mediated by signalling through production of inducible nitric oxide synthase (iNOS), a proposed effector of SWOP. Rabbits were assigned to one of four groups: (1) vehicle control; (2) MLA: (3) vehicle+aminoguanidine (AMG) control; or (4) MLA+AMG. Monophosphoryl lipid A (35 micrograms/kg) or vehicle was given intravenously 24 h before ischemia. The selective iNOS inhibitor AMG (300 mg/ kg) was injected subcutaneously 1 h before ischemia. All rabbits experienced 30 min coronary artery occlusion followed by 3 h of reperfusion. Infarct size was measured by triphenyltetrazolium chloride (TTC) staining. followed by 3 h of reperfusion. Infarct size was measured by triphenyltetrazolium chloride (TTC) staining. Myeloperoxidase activity, an index of neutrophil infiltration, was also quantified in heart tissue collected from the post-ischemic viable border zone surrounding the infarct area. MLA pretreatment significantly reduced infarct size and neutrophil infiltration in rabbit hearts compared to control (P < 0.05). Inhibition of iNOS activity by AMG abolished the infarct size reductive effect of MLA. Aminoguanidine also blocked the ability of MLA to significantly reduce neutrophil infiltration. Although measurement of iNOS activity did not show induction of the enzyme in normal myocardial tissue 24 h after MLA pretreatment, an increase in iNOS activity in ischemic tissue relative to non-ischemic tissue was found after either 15 or 30 min of coronary occlusion in MLA treated rabbits. These results suggest that MLA pretreatment may enhance iNOS enzyme activity by MLA during ischemia which may be responsible for the observed cardioprotection. PMID- 9220343 TI - Epicardial temperature is a major predictor of myocardial infarct size in dogs. AB - To determine whether epicardial temperature varies among anesthetised, open-chest dogs, and, if so, whether such variation has a measurable effect on myocardial infarct size, 35 open-chest mongrel dogs underwent 60 min of circumflex coronary artery occlusion and 3 h of reperfusion. Infarct size was measured using triphenyl tetrazolium chloride (TTC) macrochemistry. Known predictors of infarct size including area-at-risk (AAR) and collateral blood flow (CBF) were measured. Epicardial temperature was monitored using a temperature probe placed in the pericardial space adjacent to the posterior surface of the heart. In each individual dog, epicardial temperature was nearly constant throughout the period of coronary occlusion. Amongst dogs, however, epicardial temperature ranged from 35.5-41.0 degrees C. By multiple regression analysis, infarct size was better predicted by the combination of temperature and CBF than by CBF alone. "Low-T" (35.5-38.0 degrees C, n = 17) and "high-T" (38.1-41.0 degrees C, n = 18) subgroups were compared by analysis of covariance (ANCOVA), using infarct size as the dependent variable and CBF as the independent variable. Following adjustment of infarct size for CBF, infarct size in the low-T subgroup was only 53% v that in the high-T subgroup (16.9 +/- 2.7% v 31.9 +/-5.0% of AAR, P < 0.001). Thus, in open-chest dogs, relatively minor variation in epicardial temperature had major effects on myocardial infarct size. We conclude that myocardial temperature is an independent predictor of infarct size in dogs. Although such variation could confound studies of the therapeutic efficacy of proposed cardioprotective interventions, controlling for temperature variation in such studies should reduce the likelihood of false positive or negative results. PMID- 9220344 TI - Role of nuclear factor kappa B in cytokine-induced nitric oxide and tetrahydrobiopterin synthesis in rat neonatal cardiac myocytes. AB - The nitric oxide (NO) signalling pathway is thought to play a direct role in regulating the contractile properties of cardiac muscle both in vitro and in vivo. The inducible isoform of NO synthase (iNOS) mediates a sustained increase in NO production in response to cytokines in the cardiac myocytes; however, the regulation of NO synthesis in these cells remains poorly understood. Tetrahydrobiopterin (BH4) is an essential cofactor for NO formation. Cytokines induce the de novo synthesis of BH4 in cardiac myocytes, an event that is essential for the induction of NO synthesis. Activation of NO formation by cytokines in cardiac myocytes requires transcriptional induction of the genes that encode iNOS and guanosine triphosphate cyclohydrolase I (GTPCH), the first and rate-limiting enzyme in de novo BH4 synthesis. Given that nuclear factor kappa B (NF-kappa B) mediates the induction of iNOS gene expression in various cell types, the role of NF-kappa B in the induction of iNOS in cytokine stimulated rat neonatal cardiac myocytes was assessed by examining the effects of pyrrolidine dithiocarbamate (PDTC), an inhibitor of NF-kappa B activation, on the abundance of iNOS mRNA and NO synthesis. The effects of PDTC on GTPCH mRNA abundance and biopterin synthesis were also investigated. PDTC inhibited in a dose-dependent manner both NO and BH4 synthesis induced by a combination of interleukin-1 alpha (IL-1 alpha) and interferon-gamma (IFN gamma), with a half maximal inhibitory concentration of 22 muM. PDTC also prevented the accumulation of iNOS and GTPCH mRNAs induced by IL-1 alpha and IFN gamma. Cytokine-induced NO and BH4 synthesis was also inhibited by tosyl-lysine-chloromethyl ketone. another inhibitor of NF-kappa B activation. Results suggest that PDTC inhibits cytokine induced NO and BH4 synthesis by inhibiting the expression of iNOS and GTPCH genes. Thus, the induction of both genes necessary for NO synthesis in cardiac myocytes appears to be regulated, at least in part, by a common mechanism: NF kappa B activation. PMID- 9220345 TI - Regulation of phospholipases C and D in rat ventricular myocytes: stimulation by endothelin-1, bradykinin and phenylephrine. AB - The physiological activator of protein kinase C (PKC), diacylglycerol, is formed by hydrolysis of phosphoinositides (PI) by phospholipase C (PLC) or phosphatidylcholine by phospholipase D (PLD). We have measured activation of these phospholipases by endothelin-1 (ET-1), bradykinin (BK), or phenylephrine (PE) in ventricular myocytes cultured from neonatal rat. The stimulation of PI hydrolysis after 10 min by 0.1 microM ET-1 (about 12-fold) was much greater than for BK or PE (each about four-fold), and did not correlate with translocation of nPKC delta or nPKC epsilon (Clerk A. Bogoyevitch MA. Andersson MB. Sugden PH, 1994. J Biol Chem 269: 32848-32857: Clerk A, Gillespie-Brown J, Fuller SJ, Sugden PH, 1996. Biochem J 317: 109-118). However, ET-1 and BK stimulated a similar rapid increase in [3H]InsP, formation (< 30 s), which was much greater than that seen with PE. This early phase correlated with PKC translocation. Acute or chronic exposure to 12-O-tetradecanoylphorbol-13-acetate (TPA) or treatment with Ro-31-8220 showed that the stimulation of PI hydrolysis by PE, but not ET-1 or BK, was inhibited by activation of PKC. Furthermore, ET-1 and BK heterologously desensitized the stimulation of PI hydrolysis by PE, ET-1 or BK homologously uncoupled their own receptors from [3H]InsP3 formation, but there was no evidence of heterologous desensitization with these two agonists. Anomalously, chronic exposure to TPA increased the stimulation of PI hydrolysis by BK, but this probably resulted from an increase in BK receptor density. PLD was also rapidly activated by TPA. ET-1, BK or PE. Experiments with Ro-31-8220 showed that the stimulation of PLD by ET-1 and BK was mediated through activation of PKC. We discuss the characteristics of the activation of PI hydrolysis and PLD by ET-1, BK, and PE with respect to the translocation of PKC. PMID- 9220346 TI - Developmental regulation of the alpha-glycerophosphate shuttle in porcine myocardium. AB - The alpha-glycerophosphate (alpha-GP) shuttle has been shown to play a role in reducing equivalent transfer in neonatal cardiac mitochondria. In adult heart mitochondria, alpha-GP shuttle activity is not detectable. The goals of the current study were to define the time course of the age-dependent decline in alpha-GP shuttle capacity and to identify the enzymatic step(s) of the alpha-GP shuttle which are regulated during development. Intact mitochondria were isolated from porcine hearts of various ages and assayed for alpha-GP shuttle capacity. By 5 weeks of age, alpha-GP shuttle capacity had decreased by nearly 39%. The cytosolic step of the shuttle, catalysed by cytosolic alpha-glycerophosphate dehydrogenase (c alpha-GPDH), demonstrated a significant increase between 0-2-day old animals and adults. Partial cDNA clones of porcine c alpha-GPDH and mitochondrial alpha-glycerophosphate dehydrogenase (m alpha-GPDH) were prepared and used to quantitate expression of these genes. Using mRNA isolated from neonatal and adult porcine myocardium, expression of the c alpha-GPDH was unchanged, while expression of the m alpha-GPDH gene was present in neonatal but absent in adult myocardium. These results demonstrate a rapid postnatal decline in myocardial alpha-GP shuttle capacity which appears to be regulated by a decline in m alpha-GPDH gene expression. PMID- 9220347 TI - Dual effect of digitalis glycosides on norepinephrine release from human atrial tissue and bovine adrenal chromaffin cells: differential dependence on [Na+]i and [Ca2+]i. AB - It was the aim of the present study (1) to characterize the influence of Na+/K(+) ATPase inhibition by the digitalis glycoside ouabain on both spontaneous and nicotine-evoked norepinephrine release from the human heart; and (2) to further investigate the role of glycoside-induced changes in [Na+]i and [Ca2+]i (determined by microfluorimetry) for catecholamine release. The latter experiments were performed in bovine adrenal medullary chromaffin cells (BCC), an established cell culture model for sympathetic nerves. Ouabain (1-1000 mumol/l) exerted a dual effect on norepinephrine release (determined by HPLC) from incubated human atrial tissue: (I) Ouabain induced a concentration-dependent increase in norepinephrine release, that was calcium-independent and almost completely prevented by blockade of the uptake1-carrier by desipramine (1 mumol/l). The characteristics of this release process are consistent with a non exocytotic mechanism. (II) In addition, ouabain augmented the nicotine-evoked (1 100 mumol/l) calcium-dependent norepinephrine release, which can be considered to be exocytotic. Na+/K(+)-ATPase inhibition also reduced the threshold concentration of nicotine from 10 to 1 mumol/l and it delayed the rapid tachyphylaxis of its norepinephrine releasing effect in human atrial tissue. In BCC, ouabain increased [Na+]i, [Ca2+]i and [3H]-norepinephrine release in parallel. Under calcium-free conditions, not only the ouabain-induced increase in [Na+]i, but also [3H]-norepinephrine release were enhanced. The ouabain-induced [3H]-norepinephrine release was always closely related to changes in [Na+]i, indicating a key role of [Na+]i for this calcium-independent non-exocytotic norepinephrine release. In addition, pretreatment with ouabain (1 mmol/l) augmented the nicotine-evoked (0.1-10 mumol/l) increments in [Na+]i, [Ca2+]i and [3H]-norepinephrine release. As nicotine-induced norepinephrine release depends on an increase in both [Na+]i and [Ca2+]i, these findings are indicative of an ouabain-mediated facilitation of exocytosis. In conclusion, increasing [Na+]i and [Ca2+]i inhibition of Na+/K(+)-ATPase by ouabain triggers non-exocytotic norepinephrine release, and facilitates nicotine-evoked exocytotic norepinephrine release. PMID- 9220348 TI - Resting tension participates in the modulation of active tension in isolated guinea pig ventricular myocytes. AB - We studied active and passive properties of intact isolated guinea-pig ventricular myocytes in auxotonic conditions. Cells were attached using carbon fibres. The passive properties of the myocytes, in the presence of the stretch activated channel blocker streptomycin sulphate, could be separated into two groups: stiff cells (stiffness slope = 2.88 +/- 0.93 nN/micron3, n = 63 cells) and compliant cells (stiffness slope = 0.91 +/- 0.35 nN/micron3, n = 52 cells). The study and the localization of the different kind of cells indicated that endocardium is mainly constituted of stiff cells (80%) while the epicardium contained more compliant cells (60%). When a longitudinal strain was applied to compliant cells, an increase in resting tension, diastolic sarcomere length and active tension were observed. On the other hand, in stiff cells, it induced an increase in resting tension and active tension with little change of diastolic sarcomere length. In both kinds of cells, strain had no effect on Ca2+ transient amplitude and shape. Plotting active tension v diastolic sarcomere length also clearly showed two separated populations of cells, corresponding to stiff and compliant cells. The results of the two groups of cells when plotting active tension v resting tension could not be distinguished. We conclude that resting tension is an important factor in the modulation of active tension by stretch in addition to interfilament lattice spacing or sarcomere length. PMID- 9220349 TI - Detection and cellular localization of heparin-binding epidermal growth factor like growth factor mRNA and protein in human atherosclerotic tissue. AB - Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a member of the epidermal growth factor family which binds to and activates the epidermal growth factor (EGF) receptor. HB-EGF mRNA is expressed by monocytes and vascular smooth muscle cells (VSMC) in culture, and has been shown to be a potent VSMC mitogen in vitro. The aim of this study was to screen normal and human atherosclerotic arteries and SMC cultured from these arteries for expression of HB-EGF, and to determine its cellular localization in human lesions. Using the highly sensitive technique of reverse transcription polymerase chain reaction (RT PCR), we screened biopsies taken from normal human vessel walls and atherosclerotic tissue, for expression of HB-EGF mRNA. Northern blotting and RT PCR were employed to determine levels of HB-EGF gene expression in SMC, cultured from normal and atherosclerotic arteries. Cellular localization of mRNA and protein, within human atherosclerotic plaques, was assessed using in situ hybridization with 35S labelled riboprobes, and immunohistochemistry with polyclonal antibodies specific for human HB-EGF. HB-EGF mRNA was found to be expressed in human atherosclerotic lesions and in VSMC cultured from these lesions. Expression of HB-EGF could not be detected in quiescent aortic VSMC using Northern blotting, but was highly up-regulated in these cells after treatment with basic fibroblast growth factor (bFGF) for 24 h. Although HB-EGF mRNA was detected in all vascular tissue examined using RT-PCR, in situ hybridization and immunohistochemistry revealed expression of HB-EGF in small portions of diseased arteries only. Immunohistochemistry showed strong staining for macrophages in all areas of HB-EGF expression. No association of HB-EGF with SMC was observed in any of the specimens examined. In conclusion, HB-EGF, a potent mitogen for VSMC, is expressed by macrophages in human. PMID- 9220350 TI - Localisation of cardiac GS alpha in transgenic mice overexpressing GS alpha. AB - The biochemical and physiological effects of GS alpha activation are well known; however, little is known about the anatomical localisation of GS alpha in the myocardium. Knowledge of the localisation might yield insights into G protein function in heart. The utility of immunocytochemistry using immunofluorescent methods is limited in normal hearts because of the low expression of GS alpha. In order to magnify the GS alpha signal, we studied transgenic mice overexpressing myocardial GS alpha. Immunofluorescent techniques with confocal imaging using rabbit antiserum specific for GS alpha were studied in frozen sections of mouse left ventricle. GS alpha labeling appeared to be localised to the T-tubules and intercalated disks in the GS alpha overexpressing mouse hearts, whereas the control mice showed background fluorescence with diffuse faint labeling. The localisation of GS alpha to structures involved in calcium handling and membrane conductance places GS alpha at a focal point in the regulation of these key functions. PMID- 9220351 TI - Protein kinase A increases the tension cost and unloaded shortening velocity in skinned rat cardiac muscle. AB - To address controversies concerning the effect of beta-adrenergic stimulation on the rate of cross-bridge cycling in cardiac muscle, we measured ca(2+)-induced isometric tension development, unloaded shortening velocity (Vmax) and ATPase activity of demembranated (Triton X-100 skinned) rat right ventricular trabeculae before and after treatment with the catalytic subunit of protein kinase A (PKA), which is known to mimic the action of beta-adrenergic agonists in demembranated preparations. PKA treatment (1 U/microliter, 40 min) shifted the pCa-tension relation to the right from 5.41 to 5.26 at pCa50 (the [Ca2+] required for half maximal steady state tension) without changing the steepness of the pCa-tension relation and the maximum Ca(2+)-activated tension; Vmax, as determined by the slack test, was increased for a given pCa value, despite the reduced level of isometric tension. PKA treatment also shifted the pCa-ATPase activity to the right slightly from 5.47 to 5.40 at pCa50 (the [Ca2+] required for half maximal ATPase activity), but increased the ATPase activity during a given level of steady isometric tension generation, resulting in a 33% increase of the tension cost (ATPase activity/tension). All the results obtained strongly suggest that, in rat right ventricular trabeculae, beta-adrenergic stimulation may increase the rate of cross-bridge cycling by increasing the rate of cross-bridge detachment from actin through a PKA-mediated mechanism, although PKA reduces the Ca(2+) sensitivity of the contractile system. PMID- 9220352 TI - Effects of cromakalim and glibenclamide on myocardial high energy phosphates and intracellular pH during ischemia-reperfusion: 31P NMR studies. AB - ATP sensitive potassium channel (KATP) openers (e.g. cromakalim) are thought to be cardioprotective during ischemia-reperfusion, while KATP blockers (e.g. glibenclamide) may potentiate ischemia-reperfusion damage. We studied cardiac energetics and intracellular pH, by 31P magnetic resonance spectroscopy, during ischemia-reperfusion of buffer perfused, isolated rat hearts in the presence of cromakalim (10 microM) or glibenclamide (1, 10 and 50 microM). Hearts were subjected to 25 min total global ischemia at 36.5 degrees C and reperfused for 45 min. Pre-treatment with cromakalim delayed the time to ischemic contracture (19.3 +/- 1.5 min v 15.3 +/- 0.6 for control, P < 0.05) and significantly improved recovery of function at 45 min reperfusion (84 +/- 11% pre-ischemic rate pressure product (RPP) v 38 +/- 5 for control, P < 0.05). This was accompanied by an attenuation in the loss of ATP during ischemia. Pre-treatment with glibenclamide decreased the time to ischemic contracture: 16.1 +/- 0.8 min. 15.1 +/- 0.7, 12.0 +/- 1.2 (P < 0.01) and 9.5 +/- 0.9 (P < 0.001) for control, 1, 10 and 50 microM glibenclamide respectively. 50 microM glibenclamide significantly improved functional recovery at 45 min reperfusion but 1 and 10 microM were without effect; 24 +/- 6, 22 +/- 4, 29 +/- 4 and 58 +/- 7% (P < 0.05) of pre-ischemic RPP for control, 1, 10 and 50 microM glibenclamide. During ischemia, intracellular ATP was depleted more rapidly in the presence of 50 microM glibenclamide and intracellular acidosis was significantly attenuated (final pH 6.3 v 5.8 for control). 50 microM glibenclamide also decreased tissue lactate content at the end of ischemia (75 +/- 3 mumol/g dry weight v 125 +/- 18 for control, P < 0.05) and this attenuation of lactate accumulation and consequent decreased intracellular acidosis may be responsible for the cardioprotection observed under these conditions. These latter effects are unlikely to be related to glibenclamide's KATP blocking activity. This study demonstrates that blocking of myocardial KATP does not potentiate ischemia-reperfusion injury and, in addition, illustrates the important role played by intracellular acidosis in myocardial ischemia-reperfusion injury. PMID- 9220353 TI - Persistent myocardial ischemia increases GLUT1 glucose transporter expression in both ischemic and non-ischemic heart regions. AB - Persistently ischemic myocardium exhibits increased glucose uptake which may contribute to the preservation of myocardial function and viability. Little is known about the specific molecular events which are responsible for this increase in uptake. Therefore, we investigated whether myocardial ischemia induces the gene expression of the major cardiac facilitative glucose transporters, GLUT4 and GLUT1. We determined the expression of myocardial glucose transporter mRNAs and polypeptides after 6 h of regional ischemia in a dog model by semi-quantitative Northern blotting and immunoblotting. GLUT1 but not GLUT4 expression was significantly increased in both ischemic and non-ischemic regions from the experimental hearts when compared to surgical control and normal hearts. GLUT1 mRNA expression was increased 3.4-fold and GLUT1 polypeptide expression was increased 1.7-fold in ischemic hearts when compared to normal or surgical-control hearts. There were no significant regional differences in GLUT1 expression in either normal or ischemic hearts. However, there was a tendency for GLUT1 mRNA expression to be highest in the non-ischemic regions from the 6-h ischemia hearts. These findings suggest that myocardial ischemia induces a factor or factors which stimulate GLUT1 expression in non-ischemic as well as ischemic myocardial regions. Increased GLUT1 expression may play a role in augmenting glucose uptake during ischemia. PMID- 9220354 TI - Protein kinase C activity and expression in rabbit left ventricular hypertrophy. AB - Protein Kinase C (PKC) is implicated in the induction of myocardial hypertrophy. Recent studies showed an increased activity and expression of PKC in rat left ventricular hypertrophy, but we demonstrated a decreased PKC activity and content in rabbit heart failure. The present study was designed to evaluate whether these differences were due to species or model differences. PKC activity and expression were measured in a model of mild ventricular overload, induced by a 40-50% constriction of the abdominal aorta in rabbits. Left ventricular (LV) weight/body weight ratio was increased by 14, 21 and 36% after 4, 18 and 42 days of stenosis, respectively. PKC activity was significantly decreased after 18 and 42 days of stenosis in the particulate fraction of LV, but it was not modified in the cytosolic fraction leading to a significantly decreased translocation index (particulate/total activity ratio): 18.6 +/- 2.2% and 19.4 +/- 1.6% at 18 days and 42 days of aortic stenosis, respectively, compared with 25.7 +/- 2.0% and 25.8 +/- 1.2% in corresponding sham-operated rabbits (both Ps < 0.05). Similarly, PKC content, measured by immunoblotting, was not modified in the cytosolic fractions, but decreased significantly in the particulate fractions after 18 and 42 days of stenosis. These data are, thus, different from those obtained in rat LV hypertrophy showing species differences in PKC expression in hypertrophy. They also show that hypertrophy may take place without induction of PKC. PMID- 9220355 TI - Transfection with c-Ha rasEJ modulates alpha-actin and alpha 1B-adrenoceptor gene expression in vascular smooth muscle cells. AB - Co-ordinate down-regulation of smooth muscle-specific genes and acquisition of unregulated proliferative characteristics have been proposed as hallmarks of the atherosclerotic process. In the present study, we have evaluated this reciprocal relationship by examining the impact of c-Ha-rasEJ oncogene transfection on alpha smooth muscle (SM) actin and alpha 1B-adrenoceptor (ADR) gene expression in vascular (aortic) smooth muscle cells (SMCs), c-Ha-rasEJ transfection of SMCs by lipofection (LF-1) was associated with enhanced DNA synthetic rates relative to vector controls and a significant reduction in alpha-SM actin and beta/gamma actin mRNAs. Incubation of ras- and neo-LF-1 SMCs in a restrictive serum concentration (0.1%) for 72 h inhibited DNA synthesis in both cell types, but differentially influenced the pattern of alpha-actin gene expression. While neo LF-1 cells incubated in 0.1% exhibited increased alpha-SM actin mRNA levels relative to 10% serum, slight decreases in alpha-SM actin were observed in ras-LF 1 cells under the same conditions. Cyclical stretch of randomly cycling cells, seeded on a flexible elastin substrate at a rate of 100 cycles/min for 72 h, did not significantly influence the pattern of alpha-SM or beta/gamma-actin mRNA expression in neo-LF-1 or ras-LF-1 cells. Steady-state mRNA levels of alpha 1B ADR were higher in ras-LF-1 SMCs relative to neo-LF-1 cells, and stretch increased alpha 1B-ADR mRNA levels in neo-LF-1, but not ras-LF-1 cells. Stretch inhibited [1H]thymidine incorporation into DNA in both neo- and ras-LF-1 cells relative to unstretched counterparts. These results demonstrate that c-Ha-rasEJ transfection is associated with alterations in the expression of genes associated with muscle-specific functions in vascular SMCs and implicate c-Ha-ras in the regulation of phenotypic expression in SMCs. PMID- 9220356 TI - Modulation of cardiac gap junctions: the mode of action of arachidonic acid. AB - Myocytes isolated from neonatal rat hearts were grown in culture dishes. Cell pairs were selected to examine the mode of action of arachidonic acid (AA) on gap junctions. The dual voltage-clamp method was used to measure intercellular currents and determine the gap junction conductance, gj. Exposure of cell pairs to 10 microM AA produced reversible uncoupling. Pretreatment with 10 microM POCA (sodium-2-[5-(4-chlorophenyl)-pentyl]-oxirane-2-carboxylate; which inhibits mitochondrial beta-oxidation) did not prevent AA-dependent uncoupling. Thus, it seems that metabolites of beta-oxidation are not involved in AA-induced impairment of gj. Pre-exposure to 10 microM indomethacin (which blocks the cyclooxygenase pathway of the AA-cascade) had no effect on AA-dependent uncoupling. This suggests that cyclooxygenase products such as prostaglandins or thromboxanes play no role in gj modulation. Exposure to 5 microM NDGA (nordihydroguaiaretic acid; which inhibits the 5-lipoxygenase pathway) or 10 microM ETYA (5,8,11,14-eicosatetrynoic acid: which inhibits the 12- and 15 lipoxygenase pathway) led to a reversible decrease in gj. Pre-treatment with 4 BPB (4-bromophenacyl bromide: which inhibits phospholipase A2) did not prevent the effects on gj by NDGA or ETYA. This renders it unlikely that gj is regulated by eicosanoids. Also, accumulation of endogenous AA cannot be responsible for NDGA- and ETYA-dependent uncoupling. Exposure to 75 microM SKF-525A (inhibits the epoxygenase pathway) reversibly impaired gj. This is consistent with a direct action of SKF-525A on gj, but leaves open the possibility of an involvement of epoxides. The data gathered will be discussed in terms of molecular mechanisms. Due to their amphipathic character. AA, NDGA, ETYA and SKF-525A may interfere with gj by disturbing the lipid-protein interface of the cell membranes and thereby impair gap junction channels. PMID- 9220357 TI - Intermittent ischemia: energy metabolism, cellular volume regulation, adenosine and insights into preconditioning. AB - Interruption of ischemia by brief reperfusions (I/R) is better tolerated by the heart than continuous ischemia. The present study aims to determine the metabolic profiles of isolated rat hearts during intermittent ischemia, the possible cardioprotective role of adenosine and the influence of I/R on intracellular volumes, using multinuclear NMR spectroscopy. After five I/R (5/5 min) episodes, hearts paced at 5 Hz developed pressures comparable to those of hearts continuously perfused for 50 min at 37 degrees C (CP). Following the first 5 min episode of no-flow ischemia, [ADP] dropped from 72 +/- 9 to 43 +/- 5 microM (P < 0.001) and remained stable at the end of the following reperfusions, despite a 2.5-4-fold increase during each episode of 5 min ischemia. Intracellular volumes were stable during CP at a value of 2.50 +/- 0.06 ml/g dry weight, and decreased by 4, 8, and 12% after 1, 3, and 5 I/R episodes. The phosphorylation potentials decreased from 54 +/- 8 to 4 mM-1 during each period of 5 min ischemia and were 40 +/- 6 and 28 +/- 6 mM-1 after CP and I/R5, respectively. Cardiac glycogen had decreased during 50 min of CP from 103 +/- 13 to 81 +/- 9 mumol/g dry weight and lactate production was 116 +/- 15 mumol/heart. Five I/R episodes decreased glycogen to 46 +/- 7 mumol/g dry weight (P < 0.005 v CP) and increased lactate efflux to 262 +/- 31 mumol/ heart (P < 0.005 v CP). These findings suggest that a brief ischemia/reperfusion episode increases anaerobic metabolism of exogenous glucose, reduces [ADP] and induces cellular shrinkage. Administration of the adenosine receptor blocker 8-phenyl theophylline (8PT) during intermittent perfusion depressed the developed pressure to 78 +/- 7%, accentuated the decrease in phosphorylation potential (14 +/- 4 mM-1), abolished cellular shrinkage, reduced lactate efflux and blunted the decrease in ADP following the first I/R episode. In variance, no detectable changes were observed during intermittent ischemia when the ATP-sensitive potassium channel blocker glibenclamide was administered. These data demonstrate: (a) a brief episode of ischemia/reperfusion stimulates anaerobic metabolism of exogenous glucose and lowers intracellular ADP concentration: (b) adenosine receptors are partially responsible for the glycolytic stimulation during intermittent ischemia; (c) cellular shrinkage is related to the rate of glycolysis during intermittent ischemia/reperfusion. PMID- 9220358 TI - Dopamine oxidation generates an oxidative stress mediated by dopamine semiquinone and unrelated to reactive oxygen species. AB - Dopamine (100 microM, 10-30 min) inhibits/inactivates the MgATP-dependent generation of a transmembrane proton electrochemical gradient in chromaffin granule ghosts. The dopamine dependent inhibition was enhanced by adding soluble dopamine beta-monooxygenase (DBM, 0.2 U/ml) and completely prevented by ascorbate (1 mM), dithiothreitol (2 mM) and approximately 80% by the DBM inhibitor fusaric acid (10 microM). This indicates that the inhibition is caused by the dopamine semiquinone free radical generated during DBM-dependent dopamine oxidation. Catalase, superoxide dismutase or both did not prevent the inhibition, and DBM catalysed dopamine oxidation did not change the basal level of lipid peroxidation, excluding the involvement of reactive oxygen species as being responsible for the inhibition. N-ethylmaleimide-sensitive ATPase activity (i.e. the proton translocating ATPase) in the vesicle membranes was inhibited during dopamine incubation, indicating that the toxic metabolite (dopamine semiquinone) inhibits proton pumping by inhibiting the endogenous vacuolar H(+)-ATPase. As this proton pump represents the driving force for the vesicular uptake and storage of catecholamines, the dopamine dependent inhibition, if taking place in vivo, may inhibit dopamine uptake in storage vesicles in sympathetic neurons, e.g. as observed in the myopathic hamster heart. PMID- 9220361 TI - ATP content measured by 31P NMR correlates with post-ischaemic recovery of external work output in an isolated rat heart perfused with erythrocyte suspension. AB - The limited capacity of the heart to survive ischaemia presents a recognized problem during open-heart surgery. In order to provide a baseline for the study of cardioprotection we subjected a series of isolated rat hearts to periods of ischaemia of increasing duration, and sought correlations between phosphate metabolite levels as measured by 11P NMR and post-ischaemic recovery of external work output. A strong linear correlation was found between the fraction of ATP measured after reperfusion expressed as a ratio. RATP to the pre-ischaemic level, and the fractional recovery of external work output, RW: Rw = 1.06 (SE 0.27) RATP + 0.01 (SE 0.21) (Spearman rank correlation coefficient 0.72, two-tailed P value 0.006). PMID- 9220359 TI - Angiotensin I modulates Ca-transport systems in the rat heart through angiotensin II. AB - The objective of this study was to determine the effect of angiotensin I (Ang I) treatment in vivo on two major Ca-transport systems-the L-type voltage dependent calcium channel (L-VDCC) and the Na/Ca exchanger in rat heart. For our experiments we used four groups of rats, treated differently with saline, Ang I, the ACE inhibitor enalapril and/or combination of both for 6 days, every 24 h. We observed an increase in the activity, and also in mRNA expression of the Na/Ca exchanger, after repeated administration of Ang I in vivo. The maximal binding capacity of Ca-antagonist PN 200-110, which binds to the alpha 1 subunit of the L VDCC was elevated from 0.8-1.85 pg/mg protein. mRNA expression of the voltage dependent calcium channels of L-type system was also upregulated by Ang I administration, but not when enalapril was applied simultaneously with Ang I. These results demonstrate that in vivo application of the Ang I significantly modulates not only the activity, but also expression of the Na/Ca exchanger and the L-VDCC in rat hearts through angiotensin II (Ang II). Since in the in vitro experiments on the isolated cardiomyocytes, Ang II (100 nM) increased the calcium uptake after depolarization, and the AT1 receptor agonist losartan prevented this increase, we assume that this regulation might involve the AT1 receptors. PMID- 9220360 TI - Regulation of post-translationally modified microtubule populations during neonatal cardiac development. AB - Microtubules have been implicated in a number of muscle-specific functions, including sarcomerogenesis and the regulation of heart cell beating rate. Post translationally modified microtubules (MTs) have been correlated, in other cell types, with MTs of increased stability and possibly distinct functions. This study was begun in order to determine whether neonatal heart development is associated with changes in MT populations, as a prelude to assaying their role in cardiac development and function. Biochemical and morphological studies were performed on heart tissues and cells, over the developmental range from embryonic Day 19 through neonatal Day 20, as well as only fully grown adults. The specific activity of the detyrosinating enzyme, tubulin carboxypeptidase, was high in early neonatal hearts but then decreased progressively to adulthood. Levels of tyrosinated, detyrosinated, and total tubulin varied in a complex manner over the same time period, while levels of acetylated tubulin in detergent-extracted homogenates were low in all age groups. Immunofluorescence analysis of heart sections revealed non-uniform levels of Glu and acetylated tubulin between cells over that period. Cultured, neonatal day 3 myocytes exhibited much more prominent populations of both Glu and acetylated MTs than were present in the co-isolated nonmyocytes, while cell cultures from older animals showed more restricted staining for both MT types. The immunostaining pattern for 7-tubulin, a marker for MT-organizing centers, was diffuse in the cardiomyocytes throughout this period, while the staining in the non-myocytes was much more focused and punctate. These results reveal that individual MT populations are present in developing heart tissue, and may be required for specific functions during myocyte differentiation. PMID- 9220362 TI - Development and implementation of clinical pathways for stroke on a multihospital basis. AB - This study describes the development and implementation of clinical pathways for stroke on a cooperative basis by three hospitals in the same community. The participating institutions developed separate pathways which met their respective organizational needs. This process occurred within separate hospital management structures with coordination among the institutions. They employed a common set of length of stay, quality and resource variables to evaluate the impact of the pathways. PMID- 9220363 TI - Profiles of cognitive functioning in subjects with neurological disorders. AB - This report completes a four-year funded project designed to develop a data bank of Neurobehavioral Cognitive Status Examinations (NCSE) profiles of patients with neurological based disorders. Seven facilities participated in the study. Twenty categories of neurological/ psychological disorders were obtained from the NCSE profiles of 804 patients. Four categories had a sample large enough to permit inferences about the cognitive deficits related to the specific neurologic diagnosis. Including all diagnoses, the major areas of cognitive decline were auditory memory, visual memory and construction. Subjects with dementia had similar profiles but the severity varied depending upon the diagnoses of multi infarct dementia, dementia of unknown origin or Alzheimer's disease. Profiles of cognitive functioning appear to differ among patients with different neurological/psychological disorders. The study also illustrated unique cognitive deficits that occur with different neurologic disorders, and how NCSE profiles were able to identify specific areas of cognitive deficits in population of patients with similar neurologic insults. PMID- 9220364 TI - Using a health promotion model to enhance medication compliance. AB - Noncompliance with antiepileptic medication regimens is a leading cause of treatment failure. As a behavior, its causes are multifactorial and the degree to which it is exhibited varies from patient to patient. Interventions to encourage positive behavioral change need to be based on an understanding of the psychological, physiological and psychosocial aspects which explain noncompliance. Pender's Health Promotion Model provides a framework that can be used by the nurse to identify factors which interfere with the patient's ability to practice compliance and develop a treatment plan. PMID- 9220365 TI - The neuroplastic phenomenon: a physiologic link between chronic pain and learning. AB - Recent advances in the understanding of how the mind works is the result of painstaking research which has isolated tiny regions of the spinal cord and brain, and singular chemical pathways or responses to sole neurotransmitters. The true nature of the central nervous system has eluded investigators because of its fully integrated, constantly changing structure and a symphony of chemical mediators. Each sensation, thought, feeling, movement and social interaction changes the structure and function of the brain. The mere presence of another living organism can have profound effects on the mind and body through imperceptible olfactory stimuli. Neuroplasticity is a general term referring to the ability of neurons to alter their structure and function in response to internal and external stimuli. Although differences occur with aging, this is a lifelong process. Physical and chemical neuroplastic changes occur with learning, memory and chronic pain. Evidence presented supports the notion that chronic pain is a maladaptive learned phenomenon. Further evidence supports that if severe pain is allowed to persist for more than 24 hours, the neuroplastic changes associated with the development of incurable chronic pain syndromes begin to take place. Even after chronic pain is well established, new thought and behavior patterns can be learned, allowing sufferers to restore more adaptive physiologic, cognitive and behavioral patterns. PMID- 9220366 TI - Bladder management in acute care of stroke patients: a quality improvement project. AB - A follow-up evaluation of the effectiveness of bladder management in stroke patients with voiding difficulty was conducted during the period of May-August 1995. The results of this study showed 84% of all stroke patients successfully achieved urinary continence within the first month of stroke. This promising result supports the idea that uncomplicated bladder problems in the acute phase of stroke may be addressed by a management protocol using bladder scanning to estimate volume, intermittent catheterizations/post-void residual regimens, noninvasive voiding strategies and drug therapy to facilitate effective bladder emptying. PMID- 9220367 TI - Serotonergic receptors as targets for pharmacotherapy. AB - Knowledge about neurotransmitters, their receptors and their physiological effects has exploded in the past 15-20 years. Serotonin exerts diverse effects as a neurotransmitter and in nonneural tissues. The molecular sites of serotonin binding, re-uptake and autoregulation have proven to be viable targets for pharmacological intervention in a variety of disorders. PMID- 9220368 TI - Substance P receptor expression and cellular responses to substance P in prenatal rat spinal cord cells. AB - Substance P receptors (SPRs) are expressed by prenatal rat spinal cord neurons and glial cells early in their differentiation, and SPRs may mediate developmental influences in the developing spinal cord. In order to understand better early SPR expression, we quantified SPR mRNA in the rat spinal cord during prenatal development using a cDNA probe for the rat SPR in nuclease protection assays. SPR mRNA was present in the rat spinal cord at E14, the earliest stage examined, and the presence of specific binding sites for radiolabeled SP suggested that SPRs were expressed at the protein level as well. Comparisons of samples from rats at different prenatal ages showed that the relative abundance of SPR mRNA declined by about 75% from E14 through the remainder of prenatal development. Assays of the hydrolysis of phosphatidyl inositol performed on prenatal spinal cord cells in culture revealed that SP caused a small but significant stimulation. These results show that expression of SPRs is an early molecular event in the development of the rat spinal cord in vivo and that SPRs on young spinal cord cells can mediate functional responses at early developmental stages. PMID- 9220369 TI - 3 beta-OH-5 beta-pregnan-20-one enhances [3H]GABA binding in developing chick optic lobe. AB - The neurosteroids are synthesized in the CNS and act mainly through allosteric modulation of the GABAA receptor. Structure-activity relationship studies in mammalian CNS have shown that a 3 alpha-hydroxyl group and a 5 alpha-reduced A ring are striking features for their biological activity, while the 3 beta,5 beta structures as in 3 beta,5 beta-P are completely inactive. In this work we report the enhancing activity of epipregnanolone on [3H]GABA binding to its receptor sites in the chick optic lobe. Concentration-effect curves for this neurosteroid showed a concentration-dependent activity with different potencies at the three developmental stages studied, the hatching stage being the most sensitive to the steroid stimulatory effect. The displacement of a potent 3 alpha,5 alpha steroid by epipregnanolone indicated that this steroid behaved as a partial agonist of the steroid recognition site. Considering the developmental profile for steroidogenesis in avian tissues and the biological relevance of 5 beta-reduced steroids in early development, we propose that 3 or its 3 alpha-epimer, pregnanolone, instead of the potent 3 alpha,5 alpha neurosteroids, modulates GABAA receptors in the chick optic lobe during development. PMID- 9220370 TI - Molecular cloning of the dog beta 1 and beta 2 adrenergic receptors. AB - We report the isolation of the genes encoding the beta 1 and beta 2 adrenergic receptors from dog genomic DNA. Sequence analysis of both genes revealed intronless open reading frames of 473 and 415 amino acid residues, receptively. Heterologous expression of both receptors in CHO cells indicated that both receptors are functionally similar to the human homologs. Comparing the dog beta 1 and beta 2 adrenergic receptors, the beta 1 receptor appears to bind to G proteins more tightly than the beta 2 receptor. Heterologously expressed receptors provide a convenient system for evaluating novel receptor agonists and antagonists. PMID- 9220371 TI - The heat-stable enterotoxin-guanylin receptor is expressed in rat hepatocytes and in a rat hepatoma (H-35) cell line. AB - BACKGROUND/AIMS: Guanylyl cyclase C (GC-C) is an intestinal transmembrane receptor which binds both guanylin, an endogenous ligand, and Escherichia coli heat-stable enterotoxin (STa) resulting in 5'-cyclic guanosine monophosphate (cGMP) accumulation and chloride secretion. In the adult rat, there is a high basal level of GC-C expression in the intestine, but not in the liver. Increased expression of GC-C in the rat liver has been demonstrated during the perinatal period as well as with liver regeneration and during an acute phase response. The aim of this study was to identify and utilize cell culture models to further characterize the expression of GC-C in the liver. METHODS: STa binding, STa stimulated cGMP accumulation, and GC-C RNA expression by Northern analysis were determined in primary cultures of rat hepatocytes and H-35 cells, a rat hepatoma cell line, following treatment with dexamethasone and/or interleukin-6 (IL-6). RESULTS: In rat hepatocytes treated with the combination of dexamethasone and IL 6, there was an increase in STa binding, STa-stimulated cGMP accumulation, and GC C RNA expression as compared to untreated cells. In H-35 cells treated with dexamethasone alone, there was an increase in STa binding, STa-stimulated cGMP accumulation, and GC-C RNA expression as compared to untreated cells. CONCLUSION: Primary cultures of rat hepatocytes and H-35 cells can be utilized to further study upregulation of GC-C in the hepatocyte. The expression of this receptor in hepatocytes, combined with the recent demonstration of circulating guanylin, is consistent with a functional role for GC-C in the liver. PMID- 9220372 TI - Autoradiographic localization of 5-HT1E and 5-HT1F binding sites in rat brain: effect of serotonergic lesioning. AB - 5-carboxamidotryptamine (5-CT)-insensitive binding sites labelled by [3H]5 hydroxytryptamine (5-HT) in the presence of 100 nM 5-CT and 100 nM mesulergine, were examined by semi-quantitative autoradiography in rat brain. Under these conditions most of the labelled sites correspond to 5-HT1E and 5-HT1F sites. The 5-CT-insensitive binding is located mainly in cortical layer V, caudate-putamen, interpeduncular nucleus and claustrum. In cortex and caudate-putamen, a large proportion of 5-CT-insensitive sites is displaced by 250 nM sumatriptan and can be attributed to the presence of 5-HT1F receptors. A low, but significant, level of displacement by sumatriptan was observed in the choroid plexus. Lesions of serotonergic neurones by intracerebroventricular 5,7-dihydroxytryptamine injection does not significantly modify the densities of 5-HT1E or 5-HT1F binding sites. Our findings suggest that the 5-HT1F receptor has a limited distribution in rat brain, mainly located on non-serotonergic neurones. PMID- 9220374 TI - Assessing outcome in psychoanalysis and long-term dynamic psychotherapy. AB - The efficacy of psychoanalysis and long-term psychotherapy remains a fundamentally unresolved issue for lack of methodologically sound studies. This article reviews the shortcomings of prior long-term treatment research, and presents a rationale and justification of the importance of more rigorous outcome studies. An emphasis on process research is premature when efficacy remains uncertain. The modern reconceptualization of psychotherapy in terms of hermeneutic theory is discussed in relation to the empirical model. Although historically the hermeneutic perspective has served to repudiate positivism, the hermeneutic and empirical (but not positivistic) approaches to understanding information actually share common priorities. The clearest of these is that the process is ultimately evaluated and validated by the produced effect. It is argued that the recasting of psychoanalytic technique and theory according to aesthetic and pragmatic principles is not inconsistent with contemporary outcome research paradigms so long as the professed treatment objective is clearly specified in verifiable terms. The specific methodologic problems involved in extending the successful short-term psychotherapy research model to psychoanalysis are discussed. An overview of the major components of the Columbia feasibility study currently underway is presented. Finally, a number of assessment domains-for which reliable and validated instruments exist-that are thought to be relevant to outcome are reviewed. PMID- 9220375 TI - Archaic sadism. PMID- 9220373 TI - Characterization of corticotropin-releasing hormone binding sites in the human placenta. AB - The human placenta synthesizes and secretes large amounts of corticotropin releasing hormone (CRH) which has been implicated in the triggering of parturition. The placental CRH was found to act in a paracrine manner to stimulate secretion of ACTH and beta-endorphin. In view of this we sought to characterize CRH binding sites in the human placenta. The specific binding of 125I-tyrosyl-ovine CRH (125I-oCRH) to placental membranes was dependent on time, temperature, pH, divalent cations and was reversible on addition of excess oCRH. Scatchard analysis revealed a high afinity binding site with a dissociation constant of approximately 0.7 nmol/L and maximum number of binding sites approximately 44 fmol/mg protein. Disuccinimidyl suberate, a chemical cross linker, was used to covalently attach 125I-oCRH to placental membranes. The labelled placental membranes were analyzed by SDS-PAGE and autoradiography. A major radioactively labelled band with a molecular weight of 55,000 Da was identified. In this study we have identified placental binding sites for CRH with properties similar to CRH receptors described in a number of human and animal tissues and with a molecular weight similar to that of the brain CRH receptor. These binding sites may be involved in the regulation of the placental CRH/ACTH beta-endorphin axis during pregnancy and parturition. PMID- 9220376 TI - Knowing which way is up; or, why all subjectivities are not created equal. PMID- 9220377 TI - Psychoanalysis is self-centered. PMID- 9220378 TI - The psychoanalyst working in a community mental health center. PMID- 9220379 TI - Photographs, reconstructions, and the psychoanalytic task: a clinical note. PMID- 9220380 TI - The transference field and communication among therapists. PMID- 9220381 TI - Masculinity, femininity and change in psychoanalysis. PMID- 9220382 TI - Communicating with the schizophrenic superego. AB - The procedure reported here, which I have called "conversations with superegos," raises a number of important questions, both about ordinary versus psychopathological psychic structure, and about technique. There is no space to enter into a lengthy discussion, but a few brief points may be in order. First, let us consider the issue of psychic structure. I have argued that the superego is a hostile agency within the mind whose operations are essentially inimical to the patient's growth and well-being. (As an element of psychic structure, the superego can be distinguished from the ego ideal on the basis of the associated affect, that is, guilt rather than shame. Similarly, it is possible that the superego may be constructed on the basis of identifications that are different than those that may form the basis of the ego ideal). Further, I have argued that the superego is neither a force for true moral development nor a platform for the voice of the patient's "better" nature, (although it purports to be until challenged). A structure that lies, manipulates, threatens, and appears to be motivated in the end solely by its own continued existence, must certainly be considered suspect as an authority on morality. On the contrary, I see true morality originating in the patient's ego, because in that structure lies the ability to identify with others. One could object, however, that this way of viewing the matter is an artifact of working with a particular patient population. That is to say, it might be the case that in psychotic and borderline psychotic conditions, the superego does function this way, whereas in healthy or neurotic conditions it does not. In a similar vein, it could be argued that a pattern of development that was sufficiently skewed so as to result in psychosis would be more than likely to have been peopled by hostile, punitive, and essentially destructive identification figures, such as could form the basis for a pathological superego, whereas in normal or neurotic development this might not be the case. I would disagree on the following basis: I believe that the course of development is laid out by nature; that is, events like the separation individuation phase, the oral, anal, phallic, and oedipal phases occur in all of us. It is what happens in the course of that development that will lead either to normalcy or various degrees of pathology. For example, we all project. But it is whether we project an expectation that the next person we meet is a decent, civilized human being or a monster out to destroy us that causes us happiness or pain. That said, my own view of the matter is that the general principal holds for conditions other than the patients reported here, with the difference being one of degree. Where identification figures are essentially benign and foster the development of the self, then their strictures, whether moral or merely practical, would be likely to become assimilated into the ego, without the telltale restriction of reflection and autonomy together with the accompanying guilt that characterize the superego's activity, in which case, we are not talking about superegos at all. What distinguishes psychotic and neurotic development, in my view, is not the issue of whether superego operations are benign or not, but the degree to which the ego is compromised with the resulting formation of fantasy-introjects. I have found, as if it were a principal of physics, that the stronger the ego, the weaker the superego and vice versa. And when some energy of the superego is released, it appears to go directly into the ego without my intervention, though I sometimes ask the "brain's" opinion. The second issue concerns technique. In psychosis, the voice of the superego is very loud, so much so that there are times when I have had to shout to be heard. On the other hand, my patients report that the voice of the ego may be so quiet that the patient hasn't heard it at all, or hears it as a whisper. (ABSTRACT TRUN PMID- 9220383 TI - Recollections of the liberation of Buchenwald--4/11/45. PMID- 9220384 TI - Detection of parvovirus B19-specific IgM by antibody capture radioimmunoassay. AB - A solid phase IgM-capture radioimmunoassay (MACRIA) for the detection of parvovirus B19-IgM is described (Cohen et al., 1983, J. Hyg. Camb. 91, 113-130). IgM from a dilution of patients serum is 'captured' onto a solid phase coated by anti-human IgM. To determine whether any of the IgM is specific for parvovirus B19, B19 antigen is added followed by a detector system. In the MACRIA described here the detector system comprises a mouse monoclonal antibody to parvovirus B19 and a 125I-labelled anti-mouse antibody. A calibration curve derived from a standard B19 IgM serum is used to quantify B19 IgM using a single dilution of test sera. The purpose of the protocol is the diagnosis of recent acute infection with parvovirus B19. PMID- 9220385 TI - Detection of HHV-6 genotypes by in situ hybridization with variant-specific oligonucleotide probes. AB - Human herpesvirus-6 exists in two forms, HHV-6A which has not been clearly associated with any disease, and HHV-6B, the causative agent of exanthem subitum. The two variants have been distinguished by techniques such as dot blotting and restriction fragment length polymorphism of PCR products. This study aims to establish the prevalence of HHV-6A and HHV-6B in carcinoma tissues using variant specific oligonucleotide probes. A total of 73 archived carcinoma biopsies from the oral, salivary gland, larynx, breast and cervix were obtained with seven histologically normal controls. In situ hybridization was carried out with nonradioactively labelled variant-specific probes. Samples that hybridized with both variant A and B probes were subjected further to nested PCR and digested with HindIII to distinguish the variants. A hybridization signal was observed in 76.2% of oral carcinoma tissue and 75.0% of salivary gland carcinoma tissue. In contrast, only 33.3% of cervical carcinoma tissue were positive for HHV-6 DNA. A hybridization signal was noted in all 4 laryngeal carcinoma tissues studied. However, the 10 breast carcinoma tissues studied were negative, as was the histologically normal tissue. The virus possesses tumourigenic potential and demonstrates virus transactivating properties. The frequency of HHV-6 variants in certain tumours suggest a cofactorial role in multistep carcinogenesis. While PCR amplifies selectively the predominant variant in a sample, this was not seen by in situ hybridization. The in situ hybridization technique allowed the localization of both HHV-6A and HHV-6B in the nuclei of transformed regions. PMID- 9220386 TI - A single tube two compartment reverse transcription polymerase chain reaction system for ultrasensitive quantitative detection of hepatitis C virus RNA. AB - European legislation requiring polymerase chain reaction (PCR) screening of certain plasma pools to exclude contamination with hepatitis C virus (HCV) will be introduced shortly. The concentration of HCV-RNA in contaminated pools is likely to be much lower than that typically found in single donations, so the sensitivity of the PCR assays used for the screening will be critical. We describe here the development of a quantitative PCR assay for HCV-RNA with a detection limit of approximately 40 genomes/ml, which represents a sensitivity increase of 10-100 fold over that achieved by currently available assays. This approach to quantitative reverse transcription PCR (RT-PCR) may have wide applications. PMID- 9220387 TI - Direct detection of potato leafroll virus in potato tubers by immunocapture and the isothermal nucleic acid amplification method NASBA. AB - NASBA, an isothermal amplification method for nucleic acids, was applied to the detection of RNA of potato leafroll virus (PLRV) in a single enzymatic reaction at 41 degrees C. A set of primers was selected from the coat protein open reading frame sequence of PLRV to allow amplification of viral RNA. The NASBA reaction products were visualized after electrophoresis by ethidium bromide or acridine orange staining. The specificity of the amplification products was validated by Northern blot analysis with a PLRV-specific 32P-labelled oligonucleotide probe. The procedure was coupled to immunocapture of PLRV virions from tuber extracts by immobilized antibodies in microtubes. It was possible to discriminate readily by this method between uninfected and primarily PLRV-infected potato tubers. NASBA is suitable for the direct detection of PLRV in potato tubers from primarily infected plants, offering the potential to considerably simplify the inspection of seed-potatoes for virus infection. PMID- 9220388 TI - Quantitative competitive polymerase chain reaction for detection and quantification of infectious bursal disease virus cDNA and RNA. AB - A polymerase chain reaction (PCR)-based method to measure complementary DNA (cDNA) and RNA levels of infectious bursal disease virus (IBDV) was developed. Quantification was achieved by quantitative competitive PCR (QC-PCR) amplification. A competitor, a deletion mutant of the wild type IBDV cDNA, was 10 fold serially diluted and co-amplified with IBDV cDNA after being reversely transcribed from the viral RNA. After agarose gel electrophoresis, staining, and densitometric scanning, the bands on the digitized images were analyzed and quantified by computer-assisted image analysis. Complementary DNA of standard, as well as variant strains, of serotype 1 IBDV was detected and quantified using the same QC-PCR procedures. The assay could measure IBDV cDNA levels ranging from 1 microgram to 45 fg and RNA levels ranging from 9 micrograms to 45 fg. The results indicated that QC-PCR is sensitive, easy to perform, and suitable for routine quantitation of IBDV cDNA or RNA levels. PMID- 9220390 TI - An evaluation of nine commercial EIA kits for the detection of measles specific IgG. AB - Nine commercial EIAs for measles-specific IgG were compared with haemagglutination inhibition (HI) and plaque reduction neutralization (PRN). A total of 174 sera selected, to give approximately half of the sera without measles antibody by HI, were tested by all EIAs and HI. However, there was sufficient volume of only 101 samples for testing by PRN. A dilution curve of the British Standard measles antibody serum was also tested by each EIA. Assays were evaluated qualitatively against a consensus EIA result, HI and PRN: Gull, Melotest and Behring EIAs performed best. Quantitative evaluation was by assessment of the characteristics of the standard dilution curve, and by plotting differences with PRN against mean: Gull and Melotest EIAs were best. PMID- 9220389 TI - Detection of enteroviral RNA and specific DNA of herpesviruses by multiplex genome amplification. AB - A reverse transcription (RT) multiplex polymerase chain reaction (PCR) assay was developed to allow rapid, sensitive and simultaneous detection of enteroviral RNA and herpesviral DNA specific sequences in a single tube. The method involves a reverse transcription step followed by a multiplex nested PCR in which the combination of primers amplifies cDNA from enteroviruses and specific herpesviruses DNA. Nested amplification utilises primers designed to anneal into the amplification product from the first reaction. Individual viruses were then detected and differentiated by the size of their PCR products determined using ethidium bromide stained agarose gels. To exclude false negatives due to sample inhibitors an internal amplification control, a cloned fragment of DNA from Pseudorabies virus (PRV DNA) was included in the reaction mixture. Detection levels between 0.01 and 0.001 TCID50 of prototype strains of Polio and Coxsackie type B viruses and between 1 and 100 molecules of cloned-DNA of herpesviruses prototype strains were achieved. The RT multiplex PCR method proved capable of detecting enteroviral RNA or herpesviral DNA in cerebro spinal fluid (CSF) samples from patients with aetiologically well characterized encephalitis or aseptic meningitis. PMID- 9220391 TI - Detection and quantitation of HBV DNA by semi-nested PCR in donated blood: comparison with HBV serological markers. AB - To detect and quantitate hepatitis B virus (HBV) DNA, semi-nested polymerase chain reaction (PCR) method was designed for amplifying the HBV core region DNA. Cloned HBV core region DNA was used as a quantitation control, and upon electrophoresis of the semi-nested PCR product, one, two, or three bands of amplified DNA were observed using a small (< 50 mol), moderate (around 200 mol), or large (> or = 1250 mol) quantity of the template DNA, respectively. Using this semi-nested PCR method, HBV DNA was quantitated in donated blood and tested for HBV serological markers. Most of the HBV surface antigen (HBsAg) high titer samples showed three bands on the electrophoresis, indicating a high level of HBV DNA, while most of the HBsAg low titer samples showed one band, indicating a low level of HBV DNA. HBV DNA was detected in 7 out of 36 HBsAg-undetectable and anti HBc-positive samples (19.4%) but all 7 showed one band, indicating a low level of HBV DNA. In almost all of the HBV e antigen-positive samples the HBsAg titer was high, and three bands were observed indicating a high level of HBV DNA. PMID- 9220392 TI - Use of reverse transcriptase polymerase chain reaction for detection of vaccine contamination by avian leukosis virus. AB - A reverse transcriptase polymerase chain reaction (RT-PCR) for avian leukosis virus (ALV) was developed for the detection of contamination of vaccines produced in embryonated eggs and cell cultures derived from chicken. ALV is highly pathogenic and induces a wide spectrum of disease in infected animals. ALV can be divided into five subgroups (A-E). The envelope glycoprotein (env gp85) is the main antigen determinant and responsible for subgroup classification. Viral RNA of all subgroups (A-E) was isolated and amplified using three sets of primers. Subsequently, restriction endonuclease analysis confirmed the product identity and discriminated between subgroups. In specific pathogen free (SPF) eggs experimentally inoculated with ALV, viral RNA was found in allantoic fluids, as well as in vaccines spiked with different subgroups of ALV. No adventitious virus was detected in commercially available preparations. This system provides a rapid and specific in vitro method for the detection of ALV RNA as an extraneous agent and may be applied for quality control of avian vaccines. PMID- 9220393 TI - Rapid diagnosis of encephalomyocarditis virus infections in pigs using a reverse transcription-polymerase chain reaction. AB - Encephalomyocarditis virus (EMCV) is widespread and the economic losses caused by an EMCV outbreak in pig holdings and the similarity between a foot-and-mouth disease virus (FMDV) and an EMCV infection in young piglets stress the need for a rapid, specific and broad diagnostic assay. An alternative to the time-consuming seroneutralisation assay, currently used for the characterisation of EMCV, is described. An EMCV specific reverse transcription-polymerase chain reaction (RT PCR), using primers located in a conserved region of the 3D gene of the viral genome, was developed and tested on 114 different EMCV isolates. The identity of the respective amplicons was confirmed by sequencing. The potential of this assay for future diagnostic purposes was demonstrated by applying the RT-PCR on tissue samples collected from an experimentally infected piglet. PMID- 9220394 TI - Detection of specific human immunodeficiency virus type 1 Tat-TAR complexes in the presence of mild denaturing conditions. AB - Gene expression from the human immunodeficiency virus 1 (HIV-1) is greatly enhanced by binding of the virally encoded Tat protein to a 59-base RNA stem-loop structure, the Transactivation Responsive Element (TAR), located at the 5' termini of all viral transcripts. This interaction was investigated in vitro using 32P-labelled TAR and highly purified Tat in which cysteine residues were blocked by sulpitolysis (S-Tat). It is shown that specific complex formation between S-Tat and TAR can occur in the presence of urea, with urea concentrations between 5 and 6 M causing an approximately two-fold increase in the level of binding. Two conditions favoring RNA secondary structure, low temperature (0 degree C) and the presence of divalent cations (Mg2+), diminished the level of specific binding. These observations suggest that the presence of mild denaturants promoted macromolecular refolding or rearrangement in a manner that increased the number of molecules available for binding, and present a general method for studying protein/RNA interactions where analysis has been obstructed by improper protein or RNA conformation. PMID- 9220395 TI - A spot-PCR technique for the detection of phloem-limited grapevine viruses. AB - Specific amplification of genomic fragments of grapevine trichovirus A (GVA), grapevine trichovirus B (GVB) and grapevine leafroll-associated closterovirus 3 (GLRaV3) was obtained by reverse transcription-PCR on total nucleic acid solubilized from small pieces of charged nylon membrane, on which a drop of crude infected grapevine sap was spotted (spot-PCR). A thermal treatment (95 degrees for 10 degrees) of spotted sap in a buffered solution improved the release of viral template. Consistent amplification was obtained with three viruses from fragments of the same respective blots up to 1 month after spotting, while a detection threshold limit comparable with standard PCR techniques was found for this method. Duplex PCR (i.e. amplification of different viruses from a mixed infected grapevine source) was also found to be effective, since GVA and GLRaV3 were amplified by a mixture of specific primers in the same reaction. This rapid and easy sampling technique, using leaf petioles to express crude sap, may have a wide field application for screening grapevine viruses. PMID- 9220396 TI - Improvement in establishing the period of rubella virus primary infection using a mild protein denaturant. AB - A more precise determination of the date of primary rubella infection in pregnancy is obtained by application of the urea denaturation method. Two denaturation buffers containing 6 and 8 M urea were compared for the detection of rubella IgG avidity by enzyme immunoassay. The results demonstrate that IgG avidity detected by 6M urea increases more rapidly, from 16% on the third day to 51% on the 46th day after the rash, than that obtained by 8 M urea, from 13 to 36%. The specific rubella IgG avidity detected by 6 M urea denaturation buffer increased significantly in paired sera obtained at an interval of 15 days. The same samples did not show any significant increase of IgG avidity when an 8 M urea denaturation buffer was used. The use of a mild denaturant such as 6 M urea indicates the presumptive date of primary rubella infection in pregnancy within 2 months of sampling. PMID- 9220397 TI - Comparison of three luminescent assays combined with a sandwich hybridization for the measurement of PCR-amplified human cytomegalovirus DNA. AB - Identification of human tissue contaminated by cytomegalovirus is currently carried out by PCR amplification followed by measurement of the amplicons. Three luminescent detection systems undertaken after sandwich hybridization of the amplicons were compared. The sandwich hybridization takes place between a covalent linked capture probe, bound onto a plastic 96-well plate, and a biotinylated or digoxigenin-labeled detection probe. The three non-isotopic luminescent detection systems need either streptavidin-conjugated peroxydase or streptavidin-conjugated pyruvate kinase or antibodies conjugated with alkaline phosphatase. Detection of the enzymes was carried out by measurement of light emission in the presence of, respectively, luminol for peroxidase or dioxethane for alkaline phosphatase. The kinase assay was carried out not only in the presence of its substrates, ADP and phospho-enol pyruvate, but also of luciferase, which converts the produced ATP into light. The method was found to be sensitive, with the luciferase bioluminescent assay with the production of a long lasting signal. Amplicons from eight clinical samples were detected by this combination of sandwich hybridization and the three luminescent assays. The results were comparable with nested PCR for the identification of positive samples. The same correlation was obtained with 45 clinical samples using only the pyruvate kinase detection system. The high performance of these assays is given by the specificity of the sandwich hybridization combined with the sensitivity of the luminescent detection systems. PMID- 9220398 TI - Rapid identification of Australian bunyavirus isolates belonging to the Simbu serogroup using indirect ELISA formats. AB - The Bunyavirus genus, belonging to the Bunyaviridae family, is comprised of a large group of antigenically and geographically disparate arthropod-borne viruses of medical and veterinary significance. In Australia, viruses belonging to the Simbu serogroup of the Bunyavirus genus, Akabane, Tinaroo, Peaton, Aino, Douglas, Thimiri and Facey's Paddock have been isolated. In this communication we describe two indirect ELISAs, referred to as the Simbu serogroup ELISA (SG-ELISA), and the Simbu typing ELISA (ST-ELISA), for the identification of these Simbu serogroup viruses. Infected cell lysate antigens prepared from Simbu serogroup virus isolates were assessed in the SG-ELISA for reactivity with a mouse monoclonal antibody (4H9/B11/F1). The monoclonal antibody reacted strongly with all Australian members of Simbu serogroup reference viruses and is proposed for use as a serogrouping reagent for Simbu viruses. Furthermore, the ST-ELISA enabled specific identification of viruses from within this group by recognition of characteristic reaction patterns between infected cell lysate antigens and a panel of polyclonal antisera raised to Simbu serogroup viruses. PMID- 9220399 TI - A synthetic vaccinia virus promoter with enhanced early and late activity. AB - A synthetic vaccinia virus promoter (Psel) was constructed based upon sequences which increase activity of the P7.5 early/late promoter. Comparison of luciferase activity in lysates from cells infected with recombinant vaccinia viruses expressing the luciferase gene either under the control of the P7.5 promoter or Psel, demonstrated significantly enhanced activity mediated by Psel at both early and late times post infection. This promoter may be of considerable benefit in the construction of recombinant poxviruses where early foreign gene expression is important for generating a protective immune response in vaccinated animals, or in reporter/target gene expression in vitro. PMID- 9220400 TI - Comparison of PCR primer pairs in the detection of human rhinoviruses in nasopharyngeal aspirates. AB - The development of a rapid and highly sensitive PCR assay for the detection of human rhinoviruses (HRVs) in nasopharyngeal aspirates is described. Two simple and fast commercial RNA extraction methods and four primer pairs were compared. The most sensitive RNA extraction method (Ultraspec) and primer pair (A) were applied to detection of HRV RNA in 49 nasopharyngeal aspirates, of which 31 had previously been found culture-positive for HRVs. All culture-positive specimens were found positive by PCR. In addition, four of the 18 culture-negative samples were positive by PCR. Primer pair A, however, is not specific for rhinoviruses; it also amplifies enteroviruses, and thus an additional hybridization step with an HRV-specific probe is needed for group-specific diagnosis. The assay was able to detect an amount of HRV-1B RNA corresponding to 0.01 infected cells. In addition, about 50 ag (about 10 genomes) of purified HRV-1B and CBV-3 RNA still gave a signal with this primer pair. PMID- 9220402 TI - Amplification and cloning of a long RNA virus genome using immunocapture-long RT PCR. AB - A rapid and easy method was developed in order to amplify long fragments of the genome of potato virus Y (PVY), a virus possessing a 10-kb genomic RNA. The method of immunocapture-RT-PCR was adapted, by using thermostable DNA polymerases with proofreading activity and the proper buffers and cycles, to amplify almost the whole genome of PVY in two fragments (5.6 and 4.3 kb) without purifying virions nor viral RNA. Both fragments were cloned subsequently and their ends sequenced. The method is applicable to the rapid cloning and molecular characterization of the genomes of many other RNA viruses. PMID- 9220401 TI - Cationic liposome-mediated uptake of human immunodeficiency virus type 1 Tat protein into cells. AB - The human immunodeficiency virus type 1 (HIV-1) Tat protein strongly transactivates gene expression from the viral long terminal repeat (LTR) and is required for virus efficient replication. Previous studies have shown that cells scrape-loaded in the presence of purified recombinant Tat can absorb the protein in a receptor-independent fashion. Using recombinant Tat in which cysteine residues were blocked by sulfitolysis to prevent disulfide aggregation (S-Tat) we were unable to observe this phenomenon, possibly because of improper protein folding. In this study we report that the block to cellular uptake could be overcome by mixing S-Tat with a cationic liposome, Lipofectin. When mixed with Lipofectin, S-Tat effected a specific, concentration-dependent transactivation of HIV-1 LTR-directed reporter gene activity in Hela Cells. Cellular uptake was confirmed by Western blot analysis with an anti-Tat antibody. The method described utilizes cells plated in a 96-well format, requires only nanogram quantities of S-Tat protein and is much less labor-intensive than assays involving scrape-loading, making it suitable for use as a high-throughput screen for detecting Tat inhibitors. The method may have applications for the analysis of other recombinant proteins that require uptake into intact cells for determination of functionality and presents a general technique for introducing exogenous proteins into cells. PMID- 9220403 TI - Glycolytic ATP production estimated from 31P magnetic resonance spectroscopy measurements during ischemic exercise in vivo. AB - In an oxygen-depleted muscle, glycolytically produced ATP is inversely related to the ([ATP]+ creatine phosphate [PCr]) decrease because ATP, PCr, and glycolysis are virtually the only energy sources under these conditions. In particular, the onset of glycolysis or any appreciable increase in the rate of glycolytic ATP production will lead to a slower rate of ([ATP]+ [PCr]) breakdown at a given energy consumption. To quantify this relationship, endurance athletes performed isometric foot plantar flexion (20% of a test force [TF], n = 10; 50% TF, n = 5) during local arterial occlusion. Parameters of energy metabolism were measured with 31P magnetic resonance spectroscopy (31P-MRS). During exercise, [PCr] decreased to 80 +/- 10 (20% TF) and 11 +/- 4% (50% TF) of its resting concentration, and pH dropped from 7.04 +/- 0.01 to 6.98 +/- 0.10 (20% TF) and from 7.03 +/- 0.02 to 6.70 +/- 0.10 (50% TF). In both experiments, two phases of ([ATP]+ [PCr]) decrease were observed: an initial faster decrease was followed by a slower decline. The latter phase started at about the time when the pH began to drop. The difference between a line extrapolated from the slope of the initial phase and the measured ([ATP]+[PCr]) decrease was used as an estimate for glycolytically produced ATP. This estimate and pH were significantly correlated with r = -0.97 (20% TF) and r = -0.99 (50% TF). These results indicate that glycolytically produced ATP can be estimated from the ([ATP]+ [PCr]) decrease during exercise. PMID- 9220404 TI - 1H spin-lattice relaxation parameters in the length of human intestine resected for cancer. AB - 1H spin-lattice relaxation curves were acquired for samples of intestinal adenocarcinoma (B) and of uninvolved tissue at the upper (A) and lower (C) resection margin of lengths of intestine taken at surgery from 20 patients. Each sample showed a wide distribution of relaxation times with the order of 90% of the signal in a single peak at long times. Several different single-parameter relaxation times computed from discrete-exponential analysis showed that most of the relaxation times for C and B are in the upper two-thirds of the range of times for A. The mean time for the tumor is about 10% longer (with p < 0.01) than for the upper resection margin. The difference between the tumor and the lower resection margin is not significant. Distribution width parameters associated with A and C were significantly larger than those associated with the tumors. Two exponential fits indicate that the fast-relaxing component represents a smaller signal fraction for the tumor B than for A or C. PMID- 9220405 TI - Magnetic resonance imaging differentiation of adrenal masses at 1.5 T: T2 weighted images, chemical shift imaging, and Gd-DTPA dynamic studies. AB - The purpose of our study was to assess the potential role of spin-echo (SE), chemical shift, and gadolinium-enhanced magnetic resonance imaging (MRI) in the differentiation of adrenal masses. Seventy-two adrenal masses (26 nonhyperfunctioning adenomas, 16 aldosterone-secreting adenomas and 6 other different benign cortical masses, 18 pheochromocytomas, and 6 malignant masses) in 63 patients were evaluated with spin-echo sequences, chemical shift imaging (CSI) and gadolinium diethylenetriamine penta-acetic acid (Gd-DTPA) dynamic studies. Ratios and indices of signal intensity for all examined MRI methods were calculated and examined for significance of difference between different types of adrenal masses. Quantitative magnetic resonance evaluation of adrenal masses showed significant differences (at least alpha < 0.01) between nonhyperfunctioning adenomas vs. pheochromocytomas or vs. malignant lesions or vs. aldosterone-secreting adenomas and between pheochromocytomas vs. malignant lesions. The most specific indicators of adrenal mass character proved to be the CSI ratio based on opposed-phase and in-phase two-dimensional fast low-angle shot (FLASH) images, reflecting lipid content in the lesion, and Gd-DTPA dynamic studies ratios reflecting contrast agent inflow and washout in the lesion: WoMAX/LAST and Dyn1.2-3.2. There was no overlap of CSI ratio between adenomas and pheochromocytomas. The overlap of ranges of CSI ratio between nonhyperfunctioning adenomas and aldosterone-secreting adenomas was only 18.5%. There was no overlap of WoMAX/LAST ratio between adenomas and pheochromocytomas, or adenomas and malignant lesions. The overlap of ranges of Dyn1.2-3.2 ratio between pheochromocytomas and malignant lesions was only 17.6%. MRI enables good visualization and specific characterization of adrenal masses. The optimal MRI protocol for the adrenal region is presented. PMID- 9220407 TI - Young Investigator Award presentation at the 13th Annual Meeting of the ESMRMB, September 1996, Prague. A proton-electron double-resonance imaging apparatus with simultaneous multiple electron paramagnetic resonance irradiation at 10 mT. AB - The detection of free radicals in vivo is very important for the study of many physiologic and pathologic conditions. Free radicals have been implicated in a number of diseases such as ischemia, inflammation, kidney damage, and cancer. Proton-electron double-resonance imaging (PEDRI) allows the indirect detection of free radicals via the Overhauser effect. Nitroxide free radicals used for in vivo PEDRI studies present spectra with two or three lines, but most PEDRI experiments performed to date have used only single-line electron paramagnetic resonance (EPR) irradiation. There is theoretical evidence that simultaneous irradiation of multiple EPR transitions could increase the maximum achievable PEDRI enhancement. From the experimental point of view, this requires the combined use of a suitable multiple-frequency EPR source and a multiple-tuned EPR resonator. A novel radiofrequency (RF) triple-tuned loop-gap resonator for use in PEDRI has recently been developed, and dynamic nuclear polarization (DNP) data were reported. In the present study we describe a new PEDRI apparatus, equipped with a triple-tuned resonator, that is suitable for simultaneous double- or triple-EPR irradiation of nitroxide free radicals. In particular, the details of the EPR hardware used to generate the two or three EPR frequencies are given, and PEDRI images obtained with simultaneous multiple EPR irradiation are shown. Moreover, DNP experimental results showing the increase of the enhancement as a function of the EPR power for single and simultaneous double EPR irradiation are presented. The main goal of this apparatus is to improve the sensitivity and/or to reduce EPR irradiation power in a PEDRI experiment. This is likely to be particularly important in future biologic applications of PEDRI where the applied power must be optimized to reduce sample heating. PMID- 9220408 TI - TO5 as a magnetization transfer contrast test object: characterization of the gels and determination of the real magnetization transfer. AB - Following the work of the European concerted action, "Tissue characterization by magnetic resonance spectroscopy and imaging," sets of five test objects (TO) were designed, produced, and distributed among European laboratories. The TO were designed to control the image quality of clincial magnetic resonance imaging in an independent and uniform mode. The fifth test object (TO5) was devoted to relaxation measurements and composed of 18 agarose tubes, inserted in an holder filled with a CuSO4 solution. These gels are subject to magnetization transfer (MT). The purpose of this paper is to characterize their MT parameters. An individual study of each gel was performed in a spectrometer, and an individual fit, as well as a global fit, was done on the two-pool model. The MT parameters found in each case are in agreement with the known properties of the agarose gels and given below. The real MT (transfer of magnetization from water to macromolecules) was computed, taking into account the "bleeding over" (direct saturation of the water magnetization). The maximum real MT ranges from 15 to 35% and can be obtained with almost the same saturation pulse conditions for all the gels. However, the saturating field required to reach the maximum MT is very high (46 microT) and unserviceable on a clinical device. PMID- 9220406 TI - Decreasing choline signal--a marker of phenylketonuria? AB - We found a statistically significant increase in the N-acetylaspartate/choline containing compounds (NAA/Cho) ratio in a group of 69 phenylketonuria (PKU) patients with a rise in echo time (TE) compared with a group of 35 age-matched controls. The absolute concentration of creatine did not differ significantly between patients and controls, but a significant difference was found for choline containing compounds (1.33 mM in patients vs. 1.53 mM in controls, p < 0.0209). The change in NAA/Cho (for TE = 270 ms: 2.52 in patients vs. 1.96 in controls, p270 < 0.0001) can be explained by a significant difference in T2 values of choline compounds between patients and controls. This result shows that the difference in the ratios of signal intensities often used for the description of different pathologies can be explained not only by changes in the absolute metabolite concentration but also by changes in the mobility reflected by relaxation times. PMID- 9220409 TI - Magnetic resonance imaging of bovine ovaries in vitro. AB - A sample of 20 bovine ovaries were imaged in vitro using nuclear magnetic resonance (NMR) techniques to determine the visibility of various physiologic structures. In particular, the possibility of using NMR imaging to differentiate atretic follicles from physiologically selected and ovulatory follicles was examined. Five of the 20 ovaries were preserved in formalin, whereas the remaining 15 were preserved in a saline solution and imaged within 18 hours of death. Images weighted by T1 and T2 proton spin relaxation rates were obtained along with some three-dimensional (3-D) data sets acquired via a fast imaging with steady-state precession technique. Physiologically different structures were easily identified in the images from their morphology, especially in the 3-D images. Weighting by T1 and T2 was able to separate structures in the fresh ovaries in the following manner. Atretic and "cohort" follicles appear dark in T1 weighted images and bright in T2-weighted images. Ovulatory follicles appear bright in both T1- and T2-weighted images, whereas prephysiologic selection follicles present an intermediate brightness in T1-weighted images and appear dark in T2-weighted images. The corpus luteum appears bright in T1-weighted images and dark in T2-weighted images, whereas cysts in the corpus luteum appear dark in T1-weighted images and bright in T2-weighted images. The varying brightness of the follicles at different stages of development is hypothesized to be related to different hormone and protein concentrations in the follicular fluid. For example, it is known that physiologically selected preovulatory follicles contain high concentrations of estrogens in a viscous follicular fluid. The increased viscosity may occur only when the follicle fluid contains high concentrations of estrogen and contributes to bright T1-weighted images. The possibility of using nuclear relaxation-weighted NMR imaging for the study of follicular dynamics and other ovarian biology therefore shows great promise. PMID- 9220410 TI - Magnetic resonance imaging and 67Ga scan versus computed tomography in the staging and in the monitoring of mediastinal malignant lymphoma: a prospective pilot study. AB - PURPOSE: To assess the potential value of magnetic resonance imaging (MRI) combined with 67Ga single-photon emission computed tomography (SPECT) versus computed tomography (CT) in the staging and in the monitoring of mediastinal malignant lymphoma. MATERIALS AND METHODS: Twenty-three patients, referred to our institute for the evaluation of lymphoma, underwent CT, 67Ga scan, and MRI between April 1993 and February 1996 at sequential intervals. The tests studied (MRI, 67Ga, and CT) were performed according to the following schedule: 1) before start of therapy; 2) after four courses of chemotherapy; and 3) 2, 6, 12, and 18 months after the end of treatment. RESULTS: All patients studied at the time of diagnosis had abnormal gallium accumulation in the mediastinum as well as pathologic CT and pathologic signal intensity at MRI. Six months after the end of treatment full consistency was found between the results of MRI and SPECT, whereas during treatment and 2 months after the end of therapy MRI and 67Ga scan were not in agreement in nine patients. In the 23 patients in follow-up, in CT there were nine false-positive and three false-negative findings; in SPECT three false negatives; in MRI one false positive and one false negative. CONCLUSION: MRI can give morphologic information similar to CT, even superior due to multiplanarity and with major precision in the distinction between fibrosis and active disease. MRI is thus an alternative to CT. The association with SPECT allows a great diagnostic accuracy in the positive and negative predictive value. PMID- 9220412 TI - Single-shot T1- and T2-weighted magnetic resonance imaging of the heart with black blood: preliminary experience. AB - PURPOSE: To implement and evaluate two robust methods for T1- and T2-weighted snapshot imaging of the heart with data acquisition within a single heart beat and suppression of blood signal. METHODS: Both T1- and T2-weighted diastolic images of the heart can be obtained with half Fourier single-shot turbo spin echo (HASTE) and turbo fast low-angle shot (turboFLASH) sequences, respectively, in less than 350 ms. Signal from flowing blood in the ventricles and large vessels can be suppressed by a preceding inversion recovery preparing pulse pair (PRESTO). Fifteen volunteers and five patients have been evaluated quantitatively for signal-to-noise ratio (SNR) contrast-to-noise ratio (CNR) and flow void and qualitatively for image quality, artifacts, and black-blood effect. RESULTS: Both PRESTO-HASTE and PRESTO-turboFLASH achieved consistently good image quality and blood signal suppression. In contrast to gradient-echo (GRE) echo-planar imaging techniques, (EPI) HASTE and turboFLASH are much less sensitive to local susceptibility differences in the thorax, resulting in a more robust imaging technique without the need for time-consuming system tuning. Compared to standard spin-echo sequences with cardiac triggering. HASTE and turboFLASH have significantly shorter image acquisition times and are not vulnerable to respiratory motion artifacts. CONCLUSION: PRESTO-HASTE and PRESTO-turboFLASH constitute suitable methods for fast and high-quality cardiac magnetic resonance imaging (MRI). PMID- 9220411 TI - Liver and spleen enhancement after intravenous injection of carboxydextran magnetite: effect of dose, delay of imaging, and field strength in an ex vivo model. AB - It has been predicted that liver and spleen enhancement after administration of superparamagnetic contrast agents may be different, depending on the strength of the main magnetic field. With the use of an ex vivo model, we investigated at 0.3, 0.5, and 1.5 T the effects on liver and spleen signal intensity of 5, 15, and 45 mumol/kg body weight of dextran magnetite (SHU 555A) in 54 rats. Nine rats served as controls. At different time delays since injection, the animals were killed, and after perfusion with saline, the liver, brain, and spleen were fixed in formalin. The specimens were embedded in an agar gel matrix and imaged with inversion recovery T1-weighted, proton density spin echo, and T2*-weighted gradient recalled echo (GRE) sequences. At each magnetic field strength, peak liver and spleen signal loss increased with increasing dose of the contrast medium. Signal loss was significantly more conspicuous after a dose of 15 than 5 mumol/kg body weight, but not after a dose of 45 compared with 15 mumol/kg. No signal change was observed in the brain. GRE images showed higher enhancement than proton density-weighted spin echo and inversion recovery images but were noisier. The enhancement showed a plateau between 30 min and 24 hours. Only the signal decrease of the liver after a low dose of contrast medium on GRE images was significantly higher (p < 0.01) at 1.5 than at 0.5 and 0.3 T. Other differences in respect to the field strength were less significant (p < 0.05) or nonsignificant. Differences in the spleen enhancement were nonsignificant. SHU 555A at a dose of 15 mumol/kg is an efficient intracellular contrast agent for liver and spleen at low, mid, and high field strength. Proton density spin echo images are probably the sequence of choice to exploit SHU 555A contrast effects and a wide time window for imaging after its intravenous injection does exist. PMID- 9220413 TI - Sodium ion distribution in the vitreous body. AB - We have studied the nuclear magnetic resonance (NMR) relaxation behavior, and thus the dynamic properties, of the sodium ion in the vitreous body at different temperatures. The 23Na NMR spectrum exhibits a resonance, the intensity of which accounts for an ion visibility of 100%. The 23Na longitudinal and transverse relaxation times, at all temperatures but the highest, present two components, suggesting that the sodium ions are present in two states of different mobility, whose populations are in slow exchange on the NMR time scale. The correlation times and quadrupole coupling constants for the two sodium pools have been derived. The faster relaxation of a fraction of the vitreal sodium has tentatively been ascribed to the influence of the macromolecular framework of the vitreous body. The reported information may be of use for the understanding of the diagnostic applications of 23Na magnetic resonance imaging of the ocular structures. PMID- 9220414 TI - An analysis of the intracerebral ability to eliminate a nitroxide radical in the rat after administration of idebenone by an in vivo rapid scan electron spin resonance spectrometer. AB - Electron spin resonance (ESR) measurements after intracerebroventricular injection of a nitroxide radical were carried out in rats (n = 6) that received oral idebenone for 2 weeks and in control rats (n = 5), using an in vivo rapid scan ESR spectrometer. The half-life of nitroxide, which was estimated from the change in the peak height (delta M = +1) of the ESR signals from the head, was used as a marker for the elimination of the nitroxide radical. The half-life in the rats treated with idebenone was significantly shorter than it was in the controls (p < 0.05). This finding indicates that the treatment with idebenone can enhance the intracerebral-eliminating ability of the nitroxide radical. PMID- 9220415 TI - Quantification of statistical type I and II errors in correlation analysis of simulated functional magnetic resonance imaging data. AB - The potential of statistical analyses of functional magnetic resonance images using various threshold strategies in combination with correlation analysis was studied by simulating brain activation. Differences in statistical Type I (alpha) and II (beta) errors are substantial for the various thresholds. Absolute thresholds and individualized thresholds based on the assumption of a gaussian noise distribution are producing constant alpha-errors and thus do not sufficiently improve discrimination of "truly" activated pixels even for very high contrast-to-noise ratios (CNR). Only relative threshold strategies related to the maximum correlation coefficient and thus the individual data quality and activation level, i.e., a data-driven approach, can perfectly discriminate true positives, at least for CNR > 2.5. To further improve discrimination of activated and non-activated pixel in studies with lower CNR, additional prior knowledge would be necessary. From the data presented, one would also expect that the best performing threshold strategy in this simulation study would perform best under in vivo conditions. PMID- 9220416 TI - Accuracy of computed tomography and magnetic resonance imaging in staging bronchogenic carcinoma. AB - Sixty-three patients with non-small cell bronchogenic carcinoma were prospectively and independently assessed by computed tomography (CT) and magnetic resonance imaging (MRI) before surgery. Images were interpreted by four radiologists who had no knowledge of other imaging studies, except chest x-ray, and were blinded to surgical findings. The data were compared with pathologic and histologic findings. The accuracies of CT and MRI in determining tumor classification and assessing mediastinal and hilar lymph node metastases were compared. Sensitivity of CT in determining T factor was 78%, and specificity was 96%. The values for MRI were 84% and 96%, respectively. There was no significant difference between CT and MRI in staging tumors. MRI is more accurate than CT in diagnosing mediastinal invasion in staging superior sulcus tumors and complex tumors. There was no significant difference between the accuracies of CT and MRI in detecting mediastinal node metastases; the sensitivities were 82% and 90%, respectively, and specificities were 88% and 93%, respectively. PMID- 9220417 TI - Development of pancreas. AB - Pancreatic development is reviewed in man, mammals, and birds. Anatomical differences and differing topography of pancreatic excretory ducts are described in a series of mammalian species. Species differences are discussed with respect to their embryological significance. The developmental potency of the hepatopancreatic ring is stressed. Cytodifferentiation of exocrine and endocrine cells is considered. PMID- 9220418 TI - Histology of the exocrine pancreas. AB - The morphology of the exocrine secretory unit of the pancreas, i.e. the pancreatic acinus, is reviewed. The histological features of the acini and their relation with the duct system are described. The acinar three-dimensional architecture was studied by means of different ultrastructural techniques, some of which are complementary. The fine structure and morphodynamics of the acinar cells are also described. In addition, the location of the organelles in specific cytoplasmic domains and their close morphofunctional relationship with the sequential stages of secretion of the digestive enzymes are specially emphasized. Finally, morphological approaches are suggested to achieve a better comprehension of the physiological and pathological pancreatic activities whose morphodynamics need to be further elucidated or are almost totally unknown. PMID- 9220419 TI - Distribution and ultrastructure of the autonomic nerves in the mouse pancreas. AB - Peripheral innervation of the mouse pancreas was studied by scanning and transmission electron microscopy, as well as by light microscopy (cholinesterase technique). Major nerve bundles usually ran with arteries in the connective tissue septa. They gave off delicate branches that formed plexuses around arteries and arterioles. When reaching the capillaries, nerve fibers left the arterioles and formed very loose networks in the interacinar spaces. The nerves accompanying the arteries also sent off branches toward the islets of Langerhans and formed a dense plexus around the islets. A few delicate nerve fibers were also present around the pancreatic ducts. Thus, the intrapancreatic nerves formed four plexuses: perivascular, periductal, periacinar and peri-insular. The plexuses were networks of unmyelinated nerve fibers consisting of axons with varicosities and Schwann cells. Intrapancreatic ganglia were found in the interlobular connective tissue; ganglia were often closely associated to islets of Langerhans. Our findings indicate that the "interstitial cells" described by light microscopists correspond to Schwann cells. Axons in the nerve plexuses contain transmitter vesicles and therefore represent an autonomic terminal apparatus. The rich innervation of arterioles and islets suggests that neural regulation of secretory function is mediated by control of pancreatic blood flow. PMID- 9220420 TI - Morphology of the pancreatic duct system in mammals. AB - The morphology of pancreatic excretory duct segments was reviewed in mammals. The fine structure of the epithelial lining was described in intercalated ducts, intra- and extralobular ducts, and in major pancreatic ducts. Morphological characteristics of the various cell types comprising to the duct epithelium were detailed. Principal cells in the epithelial linings of interlobular and major pancreatic ducts ("Wirsungiocytes") were discussed with respect to their appearance as either clear or dark variety. In addition, the capacity of both these cell types in elaborating mucoid glycoprotein, secretions was considered and intra- and extraepithelial mucoid glands of major pancreatic ducts (ductular glands, accessory glands) was described. Finally, the wall composition of the various excretory duct segments was described. The presence of smooth muscle cells, myofibroblasts, and a peculiar periductal vascular plexus in major interlobular ducts and in main pancreatic ducts was emphasized. PMID- 9220421 TI - Microcirculation in pancreatic function. AB - The pancreas is involved in two major bodily functions: production of hormones involved in the control of carbohydrate metabolism and the production of enzymes essential to digestion. Pancreatic function is mediated by both neurological and humoral control. The major pathway for humoral control is through the circulatory system, the level of action being in the microcirculation. This introductory paper explores the need for a deeper understanding of the dynamic morphology, i.e. the actual flow patterns in the microcirculation, as a function of the physiological state and demand to complement the careful ultrastructural mapping of the microvasculature. The current state of knowledge in this field is reviewed as a basis for identifying important areas of knowledge and ignorance, and some suggestions are made as to possible procedures for further experimental studies, particularly in the microscopic observation of the dynamics of the microcirculation with special emphasis on the need for transport studies in both directions across the microvascular wall. PMID- 9220422 TI - Morphologic sites for regulating blood flow in the exocrine pancreas. AB - The exocrine pancreas has a lobular structure and an intricate capillary network supplies the lobules. Casts of these capillaries are either straight and of constant width, provided with many shallow crests, or undulating and of varying diameter, provided with bulges and deeper constrictions. The mean capillary cast diameter is 6.32 microns (SD 0.53) and 3.91 microns (SD 0.84) at constriction sites. The first type corresponds to non-fenestrated capillaries, makes 24% of capillaries and is more frequently provided with pericytes (2.7 +/- 0.9 pericytes per capillary profile). The second type corresponds to fenestrated capillaries, comprises 76% of the capillaries and is less frequently provided with pericytes (1.5 +/- 0.6 pericytes per capillary profile). The endothelial cells of capillaries regularly form intermediate junctions and microvilli and contain microtubuli and cytoplasmic filaments. Intravital observations show that capillaries are capable of contracting and narrowing the capillary lumen. This contractility is accomplished by endothelial cells both at and apart from their nuclear regions while pericytes never contracted spontaneously during our in vivo observations. The capillary diameters estimated by intravital measurements, 3.53 microns (SD 1.05), are similar to cast measurements but differ at constricted segments from cast measurements. Flow reduction shows more variability in smaller capillaries and the flow is more reduced in capillaries of 5 microns diameter to about 40% of open capillaries vs. 68% in capillaries with 7.5 microns diameter. Veins are either provided with smooth muscle sphincters or with valves. These results indicate that corrosion casting accurately shows the geometry of capillaries. However, where the capillaries are drastically constricted, they might not be filled and therefore may be underestimated during measurements. Since none of the intravital luminal constrictions are small enough to reduce flow (smaller than 1 micron luminal diameter) and because many constrictions are effective to reduce flow, we conclude that capillaries of the exocrine pancreas are always capable of maintaining continuous blood flow yet can influence blood perfusion. The presence of venous valves in association with venous sphincters constitutes a new situation concerning blood drainage regulation in the exocrine pancreas. PMID- 9220423 TI - In vivo microscopy of the exocrine pancreas. AB - Light microscopic studies of the living acinar pancreas, although limited in number, have revealed valuable information concerning dynamic aspects of microvascular and parenchymal structure and function. For example, it has been found that: 1) the living organ in anesthetized animals can be imaged with a resolution approaching the limit of the light microscope; 2) blood flow through individual capillaries in the exocrine pancreas is intermittent; 3) blood flow through these capillaries is regulated locally by smooth muscle precapillary sphincters and within individual capillaries by endothelial cells which are spontaneously contractile as well as responsive to vasoactive substances; and 4) the formation and release of zymogen granules occurs within 45-90 minutes in acinar cells stimulated with pancreozymin. This paper reviews these studies and some of the methods used to obtain them. PMID- 9220424 TI - Lymphatic system of the pancreas. AB - A network of lymphatic vessels exists within the pancreas. The majority of vessels forming this network lie in the interlobular septa of connective tissue that subdivide the pancreas into lobes and lobules. Peripheral extensions of these interlobular lymphatics can be found within the lobules, but these intralobular lymphatics are relatively sparse. In the main, the intimate relationships of these internal pancreatic lymphatics are with the blood vessels and associated connective tissue. However in random areas, both intra- and interlobular lymphatics come into close relationship with acinar cells. Rarely are there lymphatics associated with islets of Langerhans, and then only where lymphatic vessels in connective tissue septa pass close to a pancreatic lobule that contains an islet at its periphery. Intra- and interlobular lymphatics are similar in structure. Both are thin walled having an endothelial lining and a delicate component of connective tissue. The pattern of interendothelial cell contacts and the sparsity of gaps between adjacent cells suggest that fluid movement through the intracytoplasmic system of vesicles is important in lymph formation in the pancreas. However intercellular transport is also likely to occur by a dynamic process involving fluid movement through dilatations between cells from interstitium to lymphatic lumen. Both exocrine and endocrine secretions of the pancreas may enter thoracic duct lymph directly in pancreatic lymph, but in normal circumstances this route of entry is not quantitatively important. The structural relationships between lymphatics and pancreatic parenchymal cells also make clear that lymph is not a significant pathway for their secretory products. Rather, the arrangement of lymphatics in the pancreas supports the view that lymph is primarily the drainage medium for substances that, for whatever reason, enter the interstitium. In addition, the low flow of lymph compared with that of plasma lends credence to the view that lymph is not a functionally important pathway for endocrine secretions from the pancreas to reach the blood. Both structural and functional evidence suggests that the proper functioning of the lymphatic system is of critical importance in the homeostasis of the pancreas. The lymphatic system of the pancreas, like that in other organs, is essential in the removal of excess fluid from the interstitium. In this sense, the lymphatics may be considered to serve as an overflow, protective, or safety system. When the system is inadequate or its capacity is exceeded, as in inflammation of the pancreas, exocrine secretions entering the interstitium are not cleared and the proteolytic enzymes cause major damage to the tissue. This, in turn, exacerbates the edema, accentuates the inability of lymphatics to drain the fluid, and results in further damage. The fibrosis that ensues damages the lymphatics either directly or through stricture of the surrounding connective tissue. In consequence, they become inadequate at an even earlier stage in subsequent attacks of inflammation and thereby predispose to chronic and recurrent pancreatitis. The larger interlobular lymphatics formed by the junction of their tributaries emerge upon the surface of the pancreas. There they travel primarily with blood vessels and stream toward a ring of lymph nodes that intimately surrounds the pancreas. A second system of nodes extensively involved in drainage from the pancreas is related to the front and sides of the aorta from the level of the celiac trunk to the origin of the superior mesenteric artery. This second set of nodes receives lymph either directly from the pancreas or indirectly from the first echelon of nodes that rings the organ. Although there is general agreement on the disposition of the groups within these sets of nodes, confusion results from the different classifications used by various authors. These classifications range from being purely descriptive, through an alpha and num PMID- 9220425 TI - Pancreatic insulo-acinar portal systems in humans, rats, and some other mammals: scanning electron microscopy of vascular casts. AB - Scanning electron microscopy of vascular casts showed that in the mouse, rat, and guinea pig, the pancreatic endocrine islets were frequently interlobular in position and emitted insulo-venous efferent vessels directly draining into veins. In these animals, the intralobular islets, located within the exocrine lobules, issued insulo-acinar portal vessels continuous with the lobular capillaries in addition to the insulo-venous efferent vessels. In humans, monkeys, cows, pigs, dogs, cats, and rabbits, essentially all islets in the pancreas were intralobular in location and emitted the insulo-acinar portal vessels only. In man and animals examined, especially in the murine species, many lobules lacked an islet, therefore the insular control over the exocrine pancreas seemed to be effected in more or less restricted areas of lobules. PMID- 9220426 TI - Comparative analysis of insulo-acinar portal system in rats, guinea pigs, and dogs. AB - The insulo-acinar portal system in the rat, guinea pig, and dog was comparatively analyzed using corrosion casting method in scanning electron microscopy and confocal laser scanning microscopy. In all animals examined, there were three types of arterioles according to their destination: 1) the arteriole which supplied the capillary glomerulus of the islet, 2) the arterioles which directly branched out into capillaries around the acini, and 3) the arterioles which supplied the duct system. In the rat, the afferent vessel usually ended in the cortical layer of the islet and its main branches ran along this layer before giving secondary capillary branches into the deeper regions, while in the dog and guinea pig, the region where the afferent arterioles branched out into secondary capillary branches varied among individual islets. There were three types of efferent vessels of the islet: 1) the insulo-acinar portal vessels that radiated from the islet to join the capillary network in the exocrine pancreas, 2) the emissary venules of the islet, leading directly into the systemic circulation, and 3) the insulo-ductal portal vessels which drained into the peri-ductal capillary network. In the rat and guinea pig, the intralobular islets possessed both the insulo-acinar portal vessels and the emissary venules, while the interlobular islets possessed emissary venules with occasionally occurring insulo acinar portal vessels. In the dog, most of the islets were located within the lobule and possessed preferentially the insulo-acinar portal vessels. In this animal, the lobule was supplied by several microvascular units, in the center of which was located the capillary glomerulus of the islet. The peri-insular zone of the unit was mainly supplied by the insulo-acinar portal vessels, while the periphery, the tele-insular zone, was directly supplied by arterioles as well. The venules originated at the periphery of the unit. The islet in the dog had virtually no emissary venules. Confocal laser scanning microscopy of the rat islets showed that B cells occupied the core of all islets. The microvascular architecture within the rat islet appeared to be organized as to drain blood from the A and D cell area to the B cell area of the islet. PMID- 9220427 TI - Blood flow patterns in the rat pancreas: a simulative demonstration by injection replication and scanning electron microscopy. AB - Scanning electron microscopy of vascular casts prepared by arterial injections of intentionally reduced amounts of resin showed that in the rat pancreas, the casting medium fills blood capillaries in the endocrine islets more promptly than those in the exocrine lobules and secretory ducts. Furthermore, the exocrine lobules containing endocrine islets allowed a more rapid resin flow through the insulo-acinar portal route than those lobules lacking an islet. The capillaries of secretory ducts were the last portions to be filled with resin. Since the resin used in this study was as viscous as blood and injected under a physiological pressure, the microcirculatory patterns demonstrated by the present method reflect the physiological flow pattern of blood in the pancreas. PMID- 9220428 TI - Morphology of the exocrine pancreas related to pancreatitis. AB - It has been assumed in the past that pancreatic acinar cells represent an irreversible end stage in development. Consequently, when there was an increase in structures that had the morphology of ductules, the interpretation was that they were derived from the proliferation of stem cells and/or pre-existing ductular cells. Pancreatitis, however, is regressive in nature [Bockman (1984) In: Pancreatitis: Concepts and Classification. Gyr, K.E., Singer, M.V., Sarles, H., eds. Elsevier, Amsterdam, pp. 11-15]. That is, it is characterized by parenchymal destruction and loss, rather than by expansion of parenchyma. Furthermore, it was assumed that the organization of the pancreatic parenchyma is like bunches of grapes, with spheroidal acini representing the grapes, and the ductules representing the stems. Given this organization, it would be difficult to understand how regressive changes could lead to clusters of ductular structures. Investigations using three-dimensional reconstruction and retrograde injections have altered our idea of pancreatic organization. In addition to spheroidal acini, there also are other shapes, including tubular acini. Moreover, ductules do not necessarily stop when they encounter an acinus. They may emerge on the other side. Combined ductular and acinar lumina may anastomose with each other. It is now clear that pancreatic acini may undergo redifferentiation, taking on the morphology of ductules and forming tubular complexes during pancreatitis, as well as in response to pancreatic cancer, cystic fibrosis, or blockage of the ductal system. With this understanding of pancreatic architecture and morphological plasticity, it is easier to understand the changes one sees with pancreatic diseases. PMID- 9220429 TI - Angioarchitecture of the atrophic pancreas. AB - The pancreas has a complex vasculature which comprises both exocrine and endocrine structures. Copper deficiency induces highly selective acinar cell degeneration and progressive noninflammatory lipomatosis in pancreas while Langerhans islets, ducts, and nerves remain unaffected. Pancreatic vasculature was examined in rats that had dietary copper deficiency to characterize changes in the angioarchitecture of the gland. This model was used to assess the degree to which the vasculature of non-acinar components of the gland are potentially altered under conditions of exocrine atrophy. Ultrastructure of pancreas was examined by histology, enzyme histochemistry and immunohistochemistry, corrosion casting and scanning electron microscopy, in situ vascular staining, microsphere injection, biochemical analysis, and morphometry in copper-deficient rats. Results show that no acute angiopathic changes indicative of vascular disorganization accompany atrophy. Only a reduction in the complexity of the capillary beds, which normally vascularize the dense acinar parenchyma, was found. Microsphere quantitation also showed that blood flow to the lipomatous gland remains intact. Furthermore, analysis of the angioarchitecture of the atrophied pancreas supports a largely autonomous blood supply to islets and ducts. These observations support the hypothesis that while the vasculature of the atrophied gland is modified in vascular regions severely targeted by acinar necrosis, the overall structural features of the angioarchitecture are preserved. The atrophied gland thus provides an experimental model to study the vascular routes supplying islet and ductal blood flow within the complex pancreatic circulation. PMID- 9220430 TI - Three-dimensional structure of the rat pancreatic duct in normal and inflammated pancreas. AB - We observed the corrosion casts of the Wistar rats' pancreatic ducts with scanning electron microscopy (SEM), and their conventionally fixed pancreatic tissue with SEM and transmission electron microscopy (TEM). These findings revealed the following facts about the three-dimensional structure of pancreatic duct. (1) The interlobular and intralobular ducts branch like a tree, and the intercalated ducts wind and fork into two branches, although parts of the intercalated ducts anastomose with each other. The intercellular secretory canaliculi extend from the central lumina, which run straight through the center of the acini, finally approaching close to the basement membranes of acini. (2) The lumina of pancreatic ducts (i.e., the interlobular up to the intercalated ducts) are cylindric and have smooth surfaces. The luminal surface of each epithelial cell, however, is decorated by numerous microvilli and a single cilium. The length of the latter tends to be short in proportion to the diameter of pancreatic duct. Moreover the epithelial cell surfaces, which border each central lumen, have various densities of microvilli. (3) The intraductal cilium core is provided with nine microtubules, which is different from the number of microtubules encountered within the cilium core of uterine tube or bronchial epithelium. The number of microtubules in the cross-sectioned intraductal cilia decreases toward the distal portion of cilia. SEM and TEM observations on WBN/Kob rats' pancreatic ducts suggest that increased pancreatic ductal pressure causes the helical shape of the pancreatic ductal lumen. Such a helical form might also be caused by the protrusion of epithelial cell boundaries into their lumen and the hypertrophy and hyperplasia of epithelial cells, thus leading to the formation of numerous depressions equipped with elongated cilia. PMID- 9220431 TI - Ischemia and reperfusion in pancreas. AB - Ischemic diseases of heart and brain are the primary causes of mortality in industrialized nations. The ischemic injury with the consecutive reperfusion is responsible for the disturbance of microcirculation with ensuing tissue damage and organ dysfunction. Recent evidence suggests that oxygen-derived free radicals and activated polymorphonuclear leukocytes produced in ischemic tissue are instrumental in the development of ischemic cell injury. In pancreas, ischemia/ reperfusion is proposed as a potentially damaging factor accounting in part for the pathogenesis of acute pancreatitis. Apart from ischemia/reperfusion injury, the kallikrein-kinin system mediates acute inflammation associated with enhanced capillary permeability and accumulation of polymorphonuclear leukocytes, cardinal features of ischemia/reperfusion injury also in acute pancreatitis. Therefore, it seems reasonable to use bradykinin-antagonists to influence postischemic reperfusion injury of the pancreas. In the following, we describe the pathophysiology of ischemia/reperfusion injury with special reference to the pancreatic microcirculation and morphological changes as observed in a model of complete and reversible ischemia. Furthermore, we will discuss the effects of two bradykinin-antagonists (HOE 140 and CP-0597) on functional integrity of the pancreas after ischemia/ reperfusion. PMID- 9220432 TI - Postnatal development of the harderian gland in the rabbit: light and electron microscopic observations. AB - We have investigated the development of the Harderian glands of Japanese white rabbits from birth to 4 months of age. Although two types of secretory cells comprise the glandular epithelium of the pink and white lobes in fully developed glands, the time of neonatal appearance is different between the two. Cells consisting of the pink lobe first appear on the third day of life, while cells of the white appear around seventh day of life. The ultrastructure of the Harderian glands from 1-week-old rabbits resembles that of adult animals. The gland can be divided into three parts on the basis of their epithelial cell composition at the electron microscopic level. The respective parts are composed of: (1) one type of cells with large vacuoles (pink lobe), (2) one type of cells with small vacuoles (white lobe), and (3) two types of cells with large and small vacuoles (pink white mixed portion). The relative number of plasma cells per 1 mm2 is low in both pink and white lobes during early postnatal life. However, in adult animals, the white lobe has a larger number of plasma cells than the pink lobe. These results suggest the possibility that the white lobe participates in the immune system more than does the pink. PMID- 9220433 TI - Porous channels in the cuticle of the head-arrester system in dragon/damselflies (Insecta:Odonata). AB - The ultrastructure of the porous channels (PC) of the postcervical sclerite (SPC), which provides additional head fixation to the neck in adult odonates, was studied using TEM and high resolution SEM microscopy. Single chitin-protein microfibrils, about 0.14 micron thick, are arranged into channels with cylinder like shapes. The axial rod of the chitin fiber (0.04 micron thick) is located in the center of the cylinder. The orientation of the axial rods was three dimensionally demonstrated after dissolving the protein cover with NaOH. The PCs are arranged vertically to the surface and pass from the epidermal cells through all the cuticular layers to the surface of the cuticle. In the exo- and endocuticle, the PCs are usually oval in cross-section and about 0.3 micron thick. In the endocuticle, the cross-sectional area of the PCs varies widely, from 0.01-0.15 micron2. The shape of the PC is determined by the macromolecular organization of the chitin-protein microfibrils: the long axis of the channel is orientated parallel to the axis of the preferred orientation of the cuticular microfibrils. The microfibrils tend to follow the line of the channel very closely. In fractures orientated perpendicular to the surface, the PC resembles a ribbon-like construction, which was clearly demonstrated by casts. The strongly parallel orientation of PCs in the deep layers of the cuticle changes within the microtrichia (MT), and they begin to be curved. Numerous PCs pass through the microtrichium, and most of them end on its side wall. PCs usually contain channel filaments about 0.09 micron thick. Usually, a single channel contained one filament, but channels located in the deep layers of the endocuticle have from one to five single filaments. The filaments were observed in the intact cuticle and in the cuticle enzymatically treated with chitinase, while in the cuticle treated with NaOH filaments were absent. The porous channel system of the odonate arrester is interpreted as a device transporting adhesive excretions from the epidermal cells to the cuticular surface. PMID- 9220434 TI - Crystalloids in the excretory ducts of the accessory submandibular gland of the long-winged bat, Miniopterus magnator. AB - Cytoplasmic crystalloids are abundant in the excretory ducts of the accessory submandibular gland of the long-winged bat, Miniopterus magnator. The crystalloids, which always lack a membranous enclosure, may have an intricate silhouette. They consist of parallel linear densities with a 12.5 nm periodicity. These densities actually may be thin-walled tubules. In some crystalloids, intersecting subcrystalloids produce a complex pattern of decussate densities. In a few rare instances, continuities were detected between a crystalloid and a smooth-surfaced cisternal element. In other mammalian species, similar crystalloids connected to smooth endoplasmic reticulum play a role in steroid metabolism. We postulate that the ductular crystalloids in M. magnator might be involved in production of a factor that influences mating behavior. PMID- 9220435 TI - Shrinking processes in frozen sections. PMID- 9220437 TI - Microsurgery in China: a personal view. PMID- 9220436 TI - Uranyl acetate as a primary fixative for skeletal muscle. PMID- 9220438 TI - Skin expansion versus free forearm flap in forehead reconstruction. AB - The authors present their experience in surgical reconstruction of the forehead cosmetic unit, either with tissue expansion or free tissue transfer. Some of the cases underwent a full reconstruction of the entire cosmetic forehead unit en bloc performed by means of free forearm flap such as in postoncological exeresis and in post-traumatic reconstruction. The other method of choice was tissue expansion. The authors expanded the forehead unit for a giant naevus treatment in a child, and used a bilateral expansion of the forehead for tumour clearance of the middle third of the forehead. Results and problems related to the two techniques are presented and discussed. PMID- 9220439 TI - Scutuloauricularis muscle of the rabbit: a new model for free functional muscle transplantation in the mimic system. AB - There is little information on the specific process of free, functional muscle transplantation to the face trying to restore mimic function. Up to now one of the main reasons was the impossibility of functional assessment of mimic muscles as well as their transplanted substitutes. The scutuloauricularis muscle provides an ideal model for quantitative assessment of muscle function because of its circumscript tendinous insertion and is suitable for microsurgical transplantation. In a series of 26 adult, female New Zealand rabbits the anatomical and microsurgical details were studied. The scutuloauricularis muscle weighs 1.018 +/- 0.106 g, it is supplied by the auricularis rostralis artery (diameter 0.4 +/- 0.1 mm) and the auricularis rostralis vein (diameter 1.3 +/- 0.1 mm) and one oligofascicular (2-3 fascicles) branch of the facial nerve. During isometric contraction under supramaximal electrostimulation the maximal tetanic tension was 2.995 +/- 0.928 N. Histomorphometric studies in ATP-ase stained cross-sections showed 40.5 +/- 4.6% type I fibres, 32.3 +/- 5.2% type IIA fibres, and 27.2 +/- 7.1% type IIB fibres. PMID- 9220440 TI - One hundred sixty-seven thumb replantations and revascularisations: early microvascular results. AB - One hundred sixty-seven thumb replantations and revascularisations were performed from 1977 to 1987 by the Ljubljana microsurgical team. Early microvascular results of thumb replantations and early reoperations were analysed retrospectively. Age of the patients, level of traumatic amputation, mechanism of injury, use of arterial grafts, severity (total-subtotal) of amputation and occurrence of thrombosis were potential survival factors analysed with logistic regression analysis. The overall success rate for this series was 66% (72% for failures to revascularise excluded). The most frequent cause of failure was venous thrombosis. The most critical time for failure was the first 4 days after the replantation. No microvascular complication occurred later than the seventh day and no reoperation was successful later than the third day after replantation. Survival factors were studied with logistic regression analysis which showed that the model was not statistically significant. However, estimation of relative risks gave us useful but statistically uncertain information regarding the survival factors inspected. PMID- 9220441 TI - Functional results of 46 thumb replantations and revascularisations. AB - The functional results of 46 patients with isolated thumb replantations and revascularisations were evaluated in the outpatient clinic. The modified system for evaluation of reattached parts proposed by Burton was used. The system for functional evaluation of hands consisted of three major fields: socioeconomic factors, objective assessment and subjective assessment. Certain potential factors which might have influenced the functional results were analysed using Kruskal-Wallis's and Wilcoxon's sum of ranks tests. Level of amputation (P < 0.01) and mechanism of amputation (P < 0.05) significantly influenced the functional result. Age of the patients and severity of amputation (total subtotal) had no effect on the late results. Thirty-nine patients (85%) had the same employment as before injury. All the patients had economically suitable employment and 31 patients (67%) had the same manual work as before the injury. All but 8 patients experienced cold intolerance. Satisfaction with aesthetic appearance of injured hand differed between sexes: women not being pleased with the sight of their hands in 37% (3/8) and men in 8% (3/38). All patients but one would have the operation again. PMID- 9220442 TI - Inferior functional sensory regeneration after suture of sciatic neurotomy in newborns compared with mature rats. AB - It is generally believed that nerve injuries in children regenerate better than those which occur in adults. However, there are no functional experimental studies that support this belief. This study evaluates the functional regeneration of polymodal C-fibres after nerve regeneration in newborn and mature rats 3 months after unilateral sciatic nerve neurotomy and suture. The distribution of polymodal C-fibres was tested by measuring the Evans blue-stained area in the skin after antidromic nerve stimulation. In the newborn group of regenerated animals showed that functional C-fibres were present in a significantly (P < 0.05) smaller area than found in the adult group. We conclude that the functional regeneration of C-fibres is superior in mature rats compared with newborns, 3 months after regeneration. PMID- 9220443 TI - Technetium-99m human immunoglobulin (HIG): a new substance for scintigraphic detection of bone and joint infections. AB - Technetium (99m-Tc)-labelled, polyclonal human immunoglobulin (HIG) has been described as a new agent to detect local infection and inflammation. In this study, we tested 99m-Tc HIG in 55 patients with suspected chronic (n = 42) and acute (n = 13) skeletal infection. Diagnosis was proven operatively (n = 44) and clinically (n = 11), including microbiological culture tests (n = 46). A gamma camera scan was performed 4 and 24 hours after I.v. injection of 500 MBq 99m-Tc HIG. 99m-Tc-HIG scanning achieved a sensitivity of 91% and a specificity of 93%. We found one false negative and five false positive scintigraphic results in 55 patients. No clinical or biochemical side effects were encountered after 99m-Tc HIG injection. We recommend this technique especially for localisation of low grade, chronic osteomyelitis. The mechanisms and kinetics of 99m-Tc-HIG, however, are worth investigating more extensively. PMID- 9220444 TI - Influence of early fibrinolysis inhibition on thrombus formation following microvascular trauma. AB - The effect of the fibrinolysis inhibitor tranexamic acid on early thrombus formation following microvascular trauma was investigated in the central arteries of ears in 86 rabbits (in all 172 vessels), divided into four separate blind randomised studies. In the first part a common end-to-end anastomosis was done and in the last three studies a severe trauma-arteriotomy/intimectomy was performed. Parameters studied were vessel bleeding times, patency rates, weights of intraluminal thrombotic material, haematocrit and plasma fibrinolytic activity. In the first study consisting of 14 control animals and 18 animals treated with 14 mg/kg bw of tranexamic acid, end-to-end anastomosis was performed on the central artery of one ear and on the central vein of the other ear. In the second, third, and fourth studies consisting of 18, 20, and 16 control vessels and the same number of corresponding vessels in treated animals a 7-mm longitudinal arteriotomy followed by a deep 5-mm-long intimectomy was performed. The second and third treated groups were given 14 mg/kg bw of tranexamic acid 5 min and 1 h, respectively, before reflow and the fourth group 28 mg/kg bw 5 min before reflow. The difference between the second and third studies was the addition, to mimic clinical situations, of 8.5 ml saline/kg bw 2 h before reflow in the third study. In conclusion, treatment with a single clinical dose, 14 mg/kg of tranexamic acid, did not influence vessel bleeding times or thrombus formation in the anastomotic or severe trauma models and seems safe to use. Not even a double clinical dose, 28 mg/kg, influenced thrombus formation in a statistically significant way. PMID- 9220445 TI - The fatal fight for upper limb preservation in severe electrical burns. PMID- 9220446 TI - Immunological, clinical and molecular aspects of sarcoidosis. PMID- 9220447 TI - Molecular mechanisms and future uses of antiestrogens. PMID- 9220448 TI - Cyclic AMP and sympathetic neuronal programmed cell death. AB - The survival and proper functioning of sympathetic neurons are dependent on nerve growth factor (NGF). When immature sympathetic neurons are deprived of NGF, they undergo an 'active' dying process usually termed 'programmed cell death' or 'apoptosis'. This trophic factor dependence is age-related such that the cells become less dependent on NGF as they mature. Removal of NGF in immature cultures, which triggered the process of programmed cell death, resulted in a significant decrease of intracellular cAMP levels. In contrast, when these cells matured in culture and became relatively NGF independent, NGF withdrawal did not lead to a drop of cAMP levels. Pituitary adenylate cyclase-activating polypeptide (PACAP), a naturally occurring bioactive peptide structurally similar to VIP, could increase cAMP levels in these sympathetic neurons, and delay neuronal cell death resulting from NGF deprivation. These results suggest that PACAP may serve as a neurotrophic factor in sympathetic neurons. PMID- 9220449 TI - Role of oxidative stress in the manganese and 1-methyl-4-(2'-ethylphenyl)-1,2,3,6 tetrahydropyridine-induced apoptosis in PC12 cells. AB - Oxidative stress is thought to play a key role in the apoptotic death of several cellular systems, including neurons. Oxidative stress is proposed also as a mechanism of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)- and manganese (Mn)-induced neuronal death. We have recently shown that Mn and the MPTP analogue 1-methyl-4-(2'-ethylphenyl)-1,2,3,6-tetrahydropyridine (2'Et-MPTP), which is metabolized by MAO-A to 1-methyl-4-(2'-ethylphenyl)-pyridinium ion, induce apoptosis in PC12 cells. In the present study, we evaluated the effects of deprenyl and the antioxidant drugs N-acetylcysteine (NAC) and ascorbic acid (AA) on Mn- and 2'Et-MPTP-induced apoptosis in PC12 cells. Apoptosis was tested by terminal deoxynucleotidyl transferase-mediated 2'-deoxy-uridine-5'-triphosphate nick end labelling (TUNEL) technique, flow cytometry and fluorescence microscopy. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay. Mn-induced apoptosis and decrease in cell viability was inhibited by the antioxidants NAC and AA. Deprenyl failed to inhibit the above Mn effects. Neither NAC, AA nor deprenyl were able to inhibit both 2'Et-MPTP-induced apoptosis and decrease in cell viability. These results confirm that apoptosis may be an important mechanism of cell death in MPTP- and Mn-induced parkinsonism. However, an oxidative stress mechanism may be recognized, at least in vitro, only in the Mn-induced apoptosis. PMID- 9220450 TI - Autocrine regulation of apoptosis and bcl-2 expression by nerve growth factor in early differentiating cerebellar granule neurons involves low affinity neurotrophin receptor. AB - Cerebellar granule neurons produce homogenous cultures that provide a unique opportunity for quantifying the apoptosis by propidium iodide- and deoxynucleotidyl transferase-flow cytometry combined analysis and for studying its regulation by neurotrophins. Nerve growth factor (NGF) was found to promote postmitotic survival by preventing apoptosis of newly formed and early differentiated granule neurons. This regulation could be through protein bcl-2, which was underexpressed in apoptotic granule neurons and up-regulated by NGF in a dose-dependent manner. Antibodies against low affinity NGF receptors (p75NTR) mimicked the effects of NGF, suggesting that this receptor, which is transiently expressed at high levels in postmitotic granule neurons, is involved in apoptosis signaling. Since these neurons constitutively produce NGF, this is the first demonstration of an autocrine regulation of apoptosis in the CNS. Preliminary results strongly suggest that neurotrophin-3 (NT-3) and brain derived neurotrophic factor (BDNF) are also involved in the regulation of cell death, by first promoting necrosis and then protecting the remaining cells from apoptosis. In contrast, NGF may protect against two forms of cell death and act preferentially at early stages of granule neuron development. The possibility that these neurotrophins may act in parallel and/or in sequence to regulate survival of developing granule neurons through different mechanisms is discussed in the light of findings on neurotrophin and p75NTR patterns, and p75NTR/high affinity Trk receptor coexpression. PMID- 9220451 TI - Prevention of apoptotic motoneuron death in vitro by neurotrophins and muscle extract. AB - In this study, it is shown that rat motoneurons in culture are highly dependent on trophic support and die in the absence of such support via active cell death, or apoptosis. This apoptotic death occurs in their 'in vitro' life (within 24 h) and can be partially prevented by treating the cultures with neurotrophic substances. The most effective support comes from an extract prepared from embryonic chick muscle: without muscle extract, no healthy, neurite-bearing motoneurons are present, but with 0.3 and 1.2% muscle extract, their numbers increase dose-dependently. Neurotrophins, such as brain-derived neurotrophic factor (BDNF) and neurotrophin-4 (NT-4), cannot replace muscle extract and keep motoneurons alive on their own, but in the presence of muscle extract (0.3%) they have a significant effect on survival (increase up to four times, compared to 0.3% muscle extract). At the same time, they prevent apoptotic cell death. Selective motoneuron death has been implicated in the neurodegenerative disease amyotrophic lateral sclerosis (ALS). Despite strong evidence that motoneurons in ALS die via apoptosis, this has not been shown unequivocally in post mortem material. This study shows that motoneurons are very sensitive to conditions that initiate apoptosis, and that cell death can be prevented to a large degree with relatively low concentrations of neurotrophic factors. In view of the fact that several patient trials with neurotrophic factors already are underway, studies with motoneurons in culture may prove important in understanding the mechanism of cell death and the efficacy of drugs such as neurotrophic factors, but also other types of drug. PMID- 9220452 TI - Ceramide-mediated and isoquinolinesulfonamide-sensitive pathways of neuronal death: anything in common? AB - In neurons, apoptosis can be triggered by a variety of exogenous stimuli. In spite of a significant diversity in the nature of apoptotic signals, it is possible to identify points of convergence common to neuronal apoptotic processes irrespective of the nature of the signal that has initiated apoptosis. These points of convergence can be defined as a common mechanism that is activated prior to neuronal death and, if inhibited, it can prevent apoptosis. The stress activated protein kinases (SAPK) are activated in response to a variety of cellular stresses that lead to apoptosis. The SAPK-mediated apoptosis is also a ceramide-dependent processes. Recently, we reported that stress-induced neuronal apoptosis is prevented by the isoquinolinesulfonamides (IQS) H7, H8, and H9, which are known protein kinase inhibitors. We hypothesized that IQS will prevent ceramide-induced neuronal death. We observed in primary cultures of rat cerebellar granule neurons that C2-ceramide induced morphological signs of apoptosis (assayed by propidium iodide staining) and cell death. We treated cultures with C2-ceramide in the presence or absence of IQS and assessed cell death. The IQS provided neuroprotection, suggesting that ceramide-activated SAPK might play a role in IQS-sensitive neuronal death. Similarities between the ceramide-dependent and IQS-sensitive pathways indicate that they may utilize a common principle. PMID- 9220453 TI - Dopamine-melanin induces apoptosis in PC12 cells; possible implications for the etiology of Parkinson's disease. AB - The function of neuromelanin (NM), the oxidized dopamine (DA) polymer, within the DA-producing cells in the human and primate substantia nigra (SN), is still an enigma. Some studies show that the vulnerability of nigral neurons in Parkinson's disease is correlated to their toxic NM content, while others suggest that it contributes to cellular protection. We showed recently that DA, the endogenous nigral neurotransmitter, triggers apoptosis, an active program of cellular self destruction, in neuronal cultures. In the present study, we exposed cells to synthetic dopamine-melanin (DA-M) and analysed the cellular and genetic changes. We found that exposure of PC12 cells to DA-M (0.5 mg/ml for 24 h) caused 50% cell death, as indicated by trypan blue exclusion assay and 3H-thymidine incorporation. Gel electrophoresis DNA analysis of PC12 cells treated with DA-M showed the typical apoptotic DNA ladder, indicating inter-nucleosomal DNA degradation. The DNA fragmentation also was visualized histochemically in situ by DNA end-labeling staining (the TUNEL method). The FeCl2 (0.05 mM) significantly increased DA-M toxicity, while desferrioxamine, an iron chelator, totally abolished the additive toxicity of iron. The contribution of oxidative stress in this model of DA-M-induced cell death was examined using various antioxidants. In contrast to DA, inhibition of DA-M toxicity antioxidants by reduced glutathione (GSH), N-acetyl cysteine, catalase and Zn/Cu superoxide dismutase (SOD) was very limited. In conclusion, we found that DA-M may induce typical apoptotic death in PC12 cells. Our findings support a possible role of NM in the vulnerability of the dopaminergic neural degeneration in Parkinson's disease. The differential protective effect by antioxidants against toxicity of DA and DA-M may have implications for future neuroprotective therapeutic approaches for this common neurological disorder. PMID- 9220455 TI - Cellular and molecular correlates of glutamate-evoked neuronal programmed cell death in the in vitro cultures of rat hippocampal dentate gyrus. AB - An excessive neuronal stimulation through glutamate receptors is known to result in excitotoxic cell death of apoptotic (programmed) character. Granule cells of hippocampal dentate gyrus are believed to be particularly resistant to excitotoxic insults, despite the fact that pyramidal neurons of the hippocampus proper are apparently the most vulnerable brain cells. In this study, we report that neurons derived from the rat 5-day-old dentate gyrus, and maintained in vitro for 6 days, may undergo apoptosis after treatment with L-glutamate, in a dose-dependent manner-with up to 80% of neurons displaying features of programmed cell death after 24 h exposure to 0.5 mM glutamate. This conclusion is based on morphological evaluation of the cultures, nuclear staining with Hoechst 33258 and acridine orange revealing chromatin abnormalities, as well as terminal transferase labeling of DNA fragmentation. Since apoptosis is believed to be an active process involving gene expression, immunocytochemical of c-Fos and c-Jun transcription factor proteins was performed. Elevated expression of both proteins was found to follow quickly (within 1 h) after addition of glutamate. However, this effect was not dose-dependent, thus it does not provide clear correlations to the programmed cell death. In conclusion, this study reports on the establishment of a novel apoptotic model of excitotoxicity, and invites further efforts to investigate a basis for in vitro susceptibility and in vivo resistance of dentate gyrus granule cells to excitotoxic insult evoking apoptosis. PMID- 9220454 TI - Protein kinases selectively modulate apoptosis in the developing retina in vitro. AB - In the retina of newborn rats there is evidence for two mechanisms of programmed cell death. Apoptosis of ganglion cells (RGCs) following axotomy depends on protein synthesis. In contrast, inhibition of protein synthesis leads to apoptosis in the neuroblastic layer (NBL). The induction of apoptosis following translational arrest suggests that post-translational modifications of apoptosis associated proteins may be crucial to the cell death programs in the developing retina. We investigated the possible role of protein kinases upon apoptosis in retinal explants in vitro. An increase in the intracellular concentration of cAMP produced either by the adenylyl-cyclase activator forskolin (10 microM) or by 8 Br-cAMP (1 mM), prevented apoptosis induced in the NBL by inhibition of protein synthesis, but had no statistically significant effect upon RGC death. In contrast, neither 8-Br-cGMP (1 mM) nor the specific cGMP-phosphodiesterase inhibitor zaprinast (10-100 microM) had significant effects on apoptosis in the retina. The cAMP-phosphodiesterase inhibitors isobutylmethylxantine (IBMX, 0.1-1 mM) and Ro-201724 (50-200 microM) also prevented apoptosis in the NBL. The isoquinolinesulfonamide H89 (20 microM), a specific cAMP-dependent protein kinase inhibitor, partially reverted the protective effect of either forskolin or IBMX within the NBL. Neither 12-O-tetradecanoyl phorbol-13-acetate (TPA, 10 nM) nor bisindolylmaleimide (0.2-0.5 microM), respectively an activator and an inhibitor of protein kinase C had significant effects upon the retinal explants. The protein kinase inhibitor 2-aminopurine (2-AP, 10 mM) prevented apoptosis of axotomized ganglion cells and induced apoptosis in the NBL. Forskolin prevented the apoptosis induced by 2-AP in the NBL, whereas TPA had no effect. The effects of 2-AP were, however, not dependent on inhibition of protein synthesis. The data indicate that modulation of the activity of both cAMP-dependent protein kinase and several protein kinases sensitive to 2-aminopurine selectively affect apoptosis in distinct cell layers of the developing retina. PMID- 9220456 TI - Internucleosomal breakdown of the DNA of brain cortex in human spongiform encephalopathy. AB - The analysis of chromatin structure in cases of human spongiform encephalopathies could show the possible involvement of apoptosis in neuronal cell death. Genomic DNA was purified from peripheral blood lymphocytes and from a biopsy of the brain cortex in a case of Creutzfeldt-Jakob disease. Restriction fragment polymorphism with AspI, PvuII, Del and NspI showed the pattern of wild type PrP gene. The DNA purified from the brain consisted of partially degraded DNA in internucleosomal sized fragments, whereas the DNA from peripheral blood lymphocytes showed the high molecular weight of unbroken DNA. These results are consistent with the possible activation in vivo of the apoptotic endonuclease and the internucleosomal fragmentation of DNA of the brain cortex in the patient affected by Creutzfeldt-Jakob disease, suggesting the involvement of apoptosis in neuronal cell death in human spongiform encephalopathy. PMID- 9220457 TI - Role of apoptosis in the prognosis of oligodendrogliomas. AB - Prognostic factors in oligodendrogliomas are not well defined, even considering the labeling index of proliferation markers. As in other neuroepithelial tumors, the difficulty in calculating cell loss may contribute to this uncertainty. Proliferation markers Ki-67/MIB.1 and PCNA, mitoses, apoptotic nuclei, p53 and bcl-2 expression were investigated in 98 oligodendrogliomas. Apoptosis was assessed by the aspect of nuclei, by in situ end-labeling (ISEL) technique and by c-Jun immunohistochemical demonstration. The Bcl-2 also was immunohistochemically studied for its anti-apoptotic role. Mitotic index (MI), labeling index (LI) for MIB.1 and PCNA and apoptotic index (AI) were calculated and compared among themselves and with histology and survival. It was found that AI correlated with MI (p = 0.001) and was significantly higher in anaplastic than in classic oligodendrogliomas (p = 0.001). Apoptosis occurred only slightly more frequently in cases with high LIs for proliferation markers (MIB.1 and PCNA) (p = non significant) and it was definitely higher in p53-positive cases (p = 0.008). It did not correlate with bcl-2 which was poorly expressed in oligodendrogliomas, with the exception of cells with astrocytic features. Apoptotic index correlated very weakly with survival (p = 0.05); therefore, it cannot be considered a highly reliable prognostic factor in oligodendrogliomas. PMID- 9220458 TI - DNA fragmentation characteristic of apoptosis and cell loss induced by kainic acid in rabbit retinas. AB - We have examined whether in vivo exposure to the glutamate analogue, kainic acid, induces cell loss through apoptosis and/or through necrosis. The vulnerability of rabbit retinal cells was evaluated by routine histopathology. The DNA fragmentation was examined using an in situ method (TUNEL: TdT-mediated biotin dUTP nick-end labelling) and agarose gel electrophoresis of extracted retinal DNA. Retinas were examined at 30 min, and 4, 16, 24 and 36 h, and 2-5 days following the intraocular administration of 140 nmol kainic acid. Although pyknotic cells could be seen already at 30 min post-injection, TUNEL-labelled nuclei were first observed 4 h after the injection. A relatively large number of pyknotic cells and of TUNEL-labelled nuclei were still seen at 5 days post injection. Pyknotic cells were seen throughout the inner nuclear layer (mostly in the proximal half of the layer) and in the ganglion cell layer. The TUNEL labelled nuclei were almost only seen in the proximal inner nuclear layer. Analysis of DNA by electrophoresis revealed the presence of large molecular weight fragments 4 h after the injection, and of oligonucleosome-size fragments between 16 h and 2 days after the injection. The present study thus presents evidence that, in our model, the retinal cell loss induced by kainic acid is preceded, probably in most cells, by a fragmentation of DNA characteristic of apoptotic cell death. The process of cell loss following kainic acid administration was found to be relatively slow, further suggesting that a programmed type of cell death, which eventually induces apoptosis, is involved. No indication that cells were lost also through necrosis was obtained. PMID- 9220459 TI - Induction of apoptosis in cultured human retinal pigmented epithelial cells: the effect of protein kinase C activation and inhibition. AB - The presence of the non-selective protein kinase C (PKC) inhibitors, staurosporine (100 nM) and polymyxin B (100 microM) in cultured human RPE cells for more than 24 h triggers apoptotic death. Apoptosis is characterized by a diminishing number of cells, a labelling of nuclei by the TUNEL method and by observable morphological changes. An inhibitor of PKC and cyclic nucleotide dependent protein kinases, 1-(5-isoquinolinesulphonyl)-2-methyl piperazine (H-7; 100 microM), was without effect, as was the specific PKC inhibitor, calphostin C (100 nM). The PKC-activating phorbol esters, phorbol-12-myristate-13-acetate (PMA; 1 microM) and phorbol-12,13-dibutyrate (PDB; 1 microM) and the non-tumour promoting phorbol ester, 4 alpha-PMA (1 microM) were without effect, as was the diacyl glycerol analogue, 1,2-dioctanoyl-snglycerol (DOG; 10 microM). The PKC activators did not attenuate the apoptosis induced by staurosporine or polymyxin B. Furthermore, deprivation of glucose and oxygen (simulated ischemia) for 72 h induced apoptosis: this could be prevented by inclusion of 10% (v/v) foetal bovine serum (FBS) but not by a variety of PKC activators. Six PKC isoenzymes were shown to be present in RPE cells (alpha, beta 1, beta 2, delta, epsilon, E) and only the calcium-dependent cPKC levels changed after treatment with staurosporine or simulated ischaemia. Since only the less selective inhibitors of PKC induced apoptosis, it is suggested that PKC is not involved directly in the induction process of apoptosis in RPE cells. It is possible that the staurosporine and polymyxin B-induced effects of apoptosis in RPE cells are triggered by an unknown kinase-dependent pathway, but whether the 'ischaemia' induced death is related to this same process remains to be elucidated. PMID- 9220460 TI - Topographic associations between DNA fragmentation and Alzheimer's disease neuropathology in the hippocampus. AB - To identify whether the process of apoptosis bears a topographic relationship to selected aspects of Alzheimer's disease (AD) pathology, we used an in situ nick translation method (TUNEL) to map DNA fragmentation in hippocampal sections immunostained for abnormally phosphorylated tau, which exists in the neurofibrillary tangles (NFTs) and in the dystrophic neurites associated with senile plaques. To ascertain associations of DNA fragmentation with glia, TUNEL was combined with immunohistochemistry for the astrocyte marker, glial fibrillary acidic protein (GFAP), or the microglial antigen OX-42. Consistent with previous reports, the incidence of putative DNA fragmentation detected by TUNEL was much higher in the AD brain, compared to non-demented subjects. While most TUNEL positive cells did not exhibit any systematic topographic relationship to senile plaques, which were visualized by immunostain of abnormally phosphorylated tau for dystrophic neurites, DNA fragmentation was found frequently within cells containing NFTs. In hippocampal sections prepared to visualize glia, DNA fragmentation was not observed in GFAP-positive astrocytes, but some OX-42 positive microglia exhibited TUNEL signals. Other TUNEL-positive cells were found frequently in proximity to glia. The data suggest that cells compromised by the deposition of NFTs are prone to initiate the process of apoptosis. Furthermore, some glial populations appear to be apoptotic in the AD brain. PMID- 9220461 TI - GABA and NMDA in the prevention of apoptotic-like cell death in vitro. AB - We have shown recently that cerebellar granule neurons die in the absence of depolarizing concentrations of KCl through an apoptosis-like process. To study the contributions of inhibitory (gamma-aminobutyric acid; GABA) and excitatory (glutamate) neurotransmitters in the prevention of apoptotic-like cell death in cultures grown in the presence of reduced concentrations of KCl (12.5 mM), we treated these cultures either acutely or chronically with GABA, bicuculline methiodide, a GABAA receptor antagonist, N-methyl-D-aspartate (NMDA) and/or the NMDA receptor antagonist, MK-801. Cell viability was measured with fluorescein diacetate/propidium iodide (FDA/PI) and trypan blue exclusion tests. In addition, DNA fragmentation was assessed quantitatively using an in situ terminal deoxynucleotidyl transferase assay. Our results demonstrate that treatment of cerebellar granule cell cultures maintained in 12.5 mM KCl with the glutamate receptor agonist NMDA and/or bicuculline protects against cell death and reduces DNA fragmentation. In contrast, GABA potentiated cerebellar granule cell apoptosis mediated by KCl deprivation. These data indicate that signal transduction pathways activated following NMDA receptor stimulation mimic the anti-apoptotic action of high potassium in primary cultures of cerebellar granule neurons. Also, our data support an inhibitory (hyperpolarizing) role for GABA in these cultures. Collectively, the results suggest that the neurotrophic actions of NMDA on granule cells maintained in low KCl and GABA on granule cells cultured in high KCl are due to the necessity for maintaining appropriate intraneuronal calcium concentrations. PMID- 9220462 TI - Down-regulation of neurotensin receptors after ligand-induced internalization in rat primary cultured neurons. AB - When rat cultured neurons were incubated with unlabelled neurotensin (3 nM) for 1 or 24 h at 37 degrees C, the [3H]-neurotensin specific binding measured in cell homogenates was decreased to about 35 and 65% of control values, respectively. In these experiments, the decreases in binding corresponded to reductions of Bmax values without changes in the affinity. The slow neurotensin-induced receptor down-regulation is thought to result from receptor degradation since it was reduced by the lysosomotropic drugs chloroquine and methylamine and because no change in neurotensin mRNA level could be measured after the neurotensin stimulation. After their internalization, receptors slowly reappeared at the cell surface after further incubation in the absence of the peptide. Such receptor reappearance was prevented in the presence of the protein synthesis inhibitor cycloheximide and is therefore thought to result from new synthesis and not from recycling of internalized receptors. These results indicate that the neurotensin induced receptor internalization in cultured neurons is irreversible and that it is followed by a down-regulation of the receptor through a degradative process. PMID- 9220463 TI - Two modes of stimulation by ammonia of taurine release from cultured rabbit Muller cells. AB - A previous study revealed that a 10-min ('acute') treatment of cultured Muller glia with ammonium ions (further referred to as 'ammonia') at 0.5-5 mM concentration stimulated the release of newly loaded taurine (Tau) by a cAMP dependent, osmoresistant mechanism. Here we showed that a 24 h treatment of the cells with 1 mM ammonia increased both Tau release and intracellular cAMP content in a degree similar to acute treatment with 5 mM ammonia, and the effects were similarly resistant to an increase of medium tonicity by addition of 50 mM sucrose. A 65 min superfusion of the cells with a guanylate cyclase inhibitor [methylene blue (MB)], a protein kinase inhibitor (H7) and a calcium-free buffer containing 10 mM Mg2+ (OCa-10Mg) also increased Tau release and cAMP level in the cells. Acute treatment with 5 mM ammonia of cells pretreated for 24 h with 1 mM ammonia or for 65 min with MB, H7 or OCa-10Mg produced additional significant stimulation of Tau release, without further increasing the cAMP level in the cells. By contrast, a 10-min treatment with 65 mM KCl, which is a potent, cAMP independent stimulus of Tau release in untreated Muller glia, produced no further enhancement of Tau release in ammonia-, MB-, H7 or OCa-10Mg-pretreated cells. The results indicate that acute treatment with ammonia, on top of treatments that evoke Tau release associated with an increase of cAMP, produces an extra Tau release that is cAMP-independent. Tau released by this extra ammonia treatment possibly originates from a different pool than Tau liberated by the pretreatments or 65 mM KCl. PMID- 9220464 TI - Selegiline induces dopamine release through ATP-sensitive potassium channels in the rat caudate-putamen in vitro. AB - We used superfusion chambers to investigate the role of ATP-sensitive potassium (KATP) channels in dopamine (DA) release elicited by the monoamine oxidase inhibitor selegiline in the rat caudate-putamen in vitro. Selegiline (R[-] deprenyl], but not the S[+] enantiomer, concentration-dependently induced increases in extracellular concentrations of DA, with a maximal increase to 185% in comparison to basal outflow at 0.1 mM selegiline. Since in our experimental conditions exclusive MAO inhibition does not lead to an enhancement of extracellular DA levels, the effect of selegiline on DA levels seems not to be related to MAO inhibition. Butanedione (0.1 mM), a specific KATP channel blocker, also significantly enhanced extracellular DA levels in the rat caudate-putamen to approx. 260%. Selegiline only led to an additional increase of DA outflow, when added to submaximal concentrations of butanedione or tolbutamide, implying that selegiline is acting on identical sites. When the KATP channel opener cromakalim was added to the incubation medium, basal as well as butanedione-enhanced DA levels markedly decreased to about 40% when compared to baseline values. Selegiline-activated DA release was also antagonized by cromakalim. The selegiline effect was neither modulated by preincubation with the uptake inhibitor nomifensine nor by the DA agonist quinpirole and antagonist sulpiride. In conclusion these results suggest that selegiline is able to modulate KATP channels in the caudate-putamen of the rat in vitro resulting in an enhancement of striatal DA release. PMID- 9220465 TI - Neurosteroid modulation of GABAA receptors in the developing rat brain cortex. AB - The allosteric modulation of GABAA receptors in the rat brain cortex by neurosteroids was studied at different developmental stages. GABAA receptors were identified using [3H]muscimol binding to membrane preparations obtained from embryos and neonates (postnatal day 0-PN0; postnatal day 5-PN5). Data analysis disclosed a unique population of binding sites at all ages tested. An increase in the number of receptors was observed during development reaching almost adult levels at PN5. The neurosteroids pregnanolone and allopregnanolone failed to modulate [3H]muscimol specific binding in embryos and neonates, but a positive modulation was obtained in 5-day old animals. The addition of 1 microM pregnanolone induced a 3-6-fold increase [3H]muscimol affinity in PN5 (n = 3; P < 0.03), and a 2-fold increase in receptors number in adults (n = 3; P < 0.03). The differences observed in allosteric modulation during development suggest that a change occurred during the first week of life, and this change might affect GABAA receptor function. PMID- 9220466 TI - Two pathways of nitric oxide production through glutamate receptors in the rat cerebellum in vivo. AB - The effects of N-methyl-D-aspartate (NMDA), (+)-alpha-amino-3-hydroxy-5 methylisoxazole-4-propionic acid (AMPA), and trans-(+/-)-1-amino-(1S,3R) cyclopentanedicarboxylic acid (ACPD) on nitric oxide (NO) production in the cerebellum of conscious rats were investigated by measuring the levels of total NO metabolites (nitrite plus nitrate, NOx-) in dialysates obtained by in vivo microdialysis. All glutamate receptor agonists dose-dependently increased NOx- levels. Pharmacological characterization with various glutamate receptor antagonists indicated that the effects of NMDA, AMPA and ACPD are mediated by NMDA, non-NMDA, and L(+)-2-amino-3-phosphonopropionic acid (L(+)-AP-3)-sensitive metabotropic glutamate receptors, respectively. The NO synthase (NOS) inhibitors, including NG-nitro-L-arginine methyl ester (L-NAME), NG-nitro-L-arginine (L-NA), 7-nitroindazole (7-NI), and NG-monomethyl-L-arginine, inhibited NMDA-induced, but not AMPA- or ACPD-induced, increase in NOx- levels. L-Arginine enhanced NMDA induced, but not AMPA- or ACPD-induced, increase in NOx- levels. Cytochrome P-450 inhibitors, SKF525A and erythromycin, inhibited the effect of NMDA, but not AMPA or ACPD. These results suggest that AMPA and ACPD may induce NO production through a NOS-independent pathway although NMDA receptor-mediated NO production is dependent on NOS activity in the rat cerebellum in vivo. PMID- 9220467 TI - Immobilization stress reduced the expression of neurotrophins and their receptors in the rat brain. AB - Exposure to stressful events and elevated level of stress hormones are associated with impaired spatial memory and neuronal damage in the hippocampus. These neurons are considered to be maintained by neurotrophins such as nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) and trk family of neurotrophin receptors. Male Wistar rats (6 weeks old) were exposed to immobilization stress for 8 h and their brains were processed for in situ hybridization histochemistry. Exposure to long-lasting immobilization stress reduced mRNA levels for neurotrophins and their high affinity receptors in the brain, especially in the hippocampus. Our results provide, some new information that may be relevant to the pathogenesis of stress-induced disturbances of memory and learning. PMID- 9220468 TI - Induction of major histocompatibility class II antigen on microglial cells in postnatal and adult rats following intraperitoneal injections of lipopolysaccharide. AB - Microglial cells, notably the ramified form, were induced to express major histocompatibility complex (MHC) class II antigen in postnatal and adult rats given intraperitoneal injections of lipopolysaccharide (LPS). The immunoreactive microglia which occurred in cell colonies or clusters were detected immunohistochemically with the monoclonal antibody OX-6. Some of the widely distributed MHC II positive cells were round or amoeboidic located preferentially in the perivascular area. In view of the widespread occurrence of microglial cells showing OX-6 immunoreactivity which is negligible in normal animals, it is suggested that the effect of LPS on microglia in vivo is a widespread phenomenon and is independent of age. It is suggested that the endotoxin not only triggers off the immunological potentiality of these cells but also elicits the entry of some mononuclear cells into the brain parenchyma. PMID- 9220469 TI - Labeling of amoeboid microglial cells and intraventricular macrophages in fetal rats following a maternal injection of a fluorescent dye. AB - Amoeboid microglial cells (AMC) in fetal brains were labeled by rhodamine B isothiocyanate (RhIc) when injected intravenously or intraperitoneally into mother rats at late state of pregnancy. The fluorescent cells were immunostained with antibodies OX-42 and OX-18 that recognize complement type 3 (CR3) receptors and major histocompatibility complex class I (MHC-I) surface antigen, respectively. RhIc-labeled AMC were first observed in the cavum septum pellucidum and subependymal cysts associated with the cerebral aqueduct as well as the fourth ventricle, and subsequently at other sites including the corpus callosum and other subcortical white matter. The fluorescence intensity increased with time after RhIc administration so that after 1 day the cells were brightly labeled. The majority of the labeled cells were round, with some elongated ones bearing two or three processes. Besides AMC, macrophages in the ventricular system were also labeled. All fluorescent cells were double labeled with OX-42 and OX-18 antibodies. Present results suggest that when introduced into the maternal circulation, RhIc could readily gain access into the fetal brain through the inefficient placental, blood-brain and blood-cerebrospinal-fluid (blood-CSF) barriers. The avid uptake of RhIc in circulation by brain macrophages indicates an active scavenging role of these cells in fetal brain. The labeling of cells by maternal route offers a rapid method for study of distribution of brain macrophages in fetuses. PMID- 9220470 TI - Connections of the superior colliculus with the tegmentum and the cerebellum in the hedgehog tenrec. AB - Different tracer substances were injected into the superior colliculus (CoS) in order to study its afferents and efferents with the meso-rhombencephalic tegmentum, the precerebellar nuclei and the cerebellum in the Madagascan hedgehog tenrec. The overall pattern of tectal connectivity in tenrec was similar to that in other mammals, as, e.g. the efferents to the contralateral paramedian reticular formation. Similarly the origin of the cerebello-tectal projection in mainly the lateral portions of the tenrec's cerebellar nuclear complex corresponded to the findings in species with little binocular overlap. In comparison to other mammals, however, the tenrec showed a consistent projection to the ipsilateral inferior olivary nucleus, in addition to the classical contralateral tecto-olivary projection. The tenrec's CoS also appeared to receive an unusually prominent monoaminergic input particularly from the substantia nigra, pars compacta. There was a reciprocal tecto-parabigeminal projection, a distinct nuclear aggregation of parabigeminal neurons, however, was difficult to identify. The dorsal lemniscal nucleus did not show perikaryal labeling in contrast to the paralemniscal region. Similar to the cat but unlike the rat there were a few neurons in the nucleus of the central acoustic tract. Unlike the cat, but similar to the rat there was a distinct, predominantly ipsilateral projection to the magnocellular reticular field known to project spinalward. PMID- 9220471 TI - Large dorsal horn neurons which receive inputs from numerous substance P-like immunoreactive axon terminals in the laminae I and II of the chicken spinal cord. AB - Large neurons outlined with numerous substance P (SP)-like immunoreactive (LI) boutons were detected immunocytochemically in the dorsal horn of the chicken spinal cord at the light microscopic level. The cervical enlargement was mainly used for observations. By electron microscopy, asymmetrical synapses were observed between the SP-LI axon terminals and the soma and dendrites of the large neurons. Cell bodies of the large neurons were mostly localized in the lamina I and the region lateral to the lamina I. Some of the cell bodies were also located in the lamina II. Their dendrites extended in the lamina I, in the region lateral to the lamina I, and deeply in the lamina II. In the lamina II, dendrites of these neurons formed synapses with SP-containing central terminals in synaptic glomeruli known to originate from primary afferents. The findings suggest that these large neurons receive nociceptive information directly from primary afferents. In the light of previous investigations, these neurons are considered to be pain-transmitting long ascending tract neurons. PMID- 9220472 TI - Single-unit activity in the primate nucleus tegmenti pedunculopontinus related to voluntary arm movement. AB - In the pedunculopontine tegmental nucleus (PPN), single-unit activity was recorded in two monkeys trained to manipulate an on-off lever with a hand. Among 280 neurons recorded, a change in the firing rate related to the lever-off movement was observed in 125 neurons for the contralateral limb movement (53%) and in 96 neurons for the ipsilateral limb movement (48%). The changes were an increase in the firing rate in 122 neurons and a decrease in 99 neurons. These changes in the firing rate related to the task often occurred for both the contralateral and ipsilateral limb movements. The change of activity preceded the movement onset for both contralateral and ipsilateral arm movements. These findings suggest that in primates the PPN contributes to coordination of upper limb movements on both sides. PMID- 9220473 TI - Response of glial cells and activation of complement following motorneuron degeneration induced by toxic ricin. AB - Motor nerve transection in adult rats induce a series of metabolic and structural changes in the injured neurons as well as in surrounding glial cells; however, without substantial neuronal degeneration. In the present study we found, in contrast with axotomy, a massive neuronal death in the ipsilateral hypoglossal nucleus following injection of toxic ricin (RCA) into the hypoglossal nerve, which is in line with previous observations. Injection of RCA enables examination of the glial reaction in a situation where neuronal degeneration is profound, which has been the approach in the present study. We found an increase in OX42-, GFAP-, and transferrin-immunoreactivity in microglial, astroglial, and oligodendroglial cells respectively, in the ipsilateral hypoglossal nucleus three to seven days following injection of toxic ricin in the hypoglossal nerve. Proliferation was found in astrocytes as well as in microglial cells, as shown by uptake of bromodeoxyuridine. In addition, the complement cascade was activated locally in the ipsilateral hypoglossal nucleus, as demonstrated by immunohistochemical detection of complement components C3d and C9. Complement activation may serve several effects in the glial-neuronal interactions. Stimulation of phagocytosis by reactive microglia is probably the most important one. Furthermore, the degenerative neuronal somata showed increased immunoreactivity for clusterin, which is a known complement inhibitor, but a decrease in clusterin-mRNA. In conclusion, the glial cell response was in several aspects principally different following massive motorneuron degeneration induced by toxic ricin in comparison to previous findings reported after axotomy. PMID- 9220474 TI - Prominent expression of nuclear hormone receptor ROR alpha in Purkinje cells from early development. AB - The ROR alpha is a member of the nuclear hormone receptor gene superfamily, and its deletion causes the staggerer mutation in mice. In the staggerer mutant mouse, Purkinje cells (PCs) are severely affected in the cytology, synapse formation and gene expression. We previously found the presence of mediolateral compartments unique to the staggerer cerebellum, based on different degrees of abnormalities in the cytology and gene expression. In this paper we investigated expression of the ROR alpha mRNA in developing mouse cerebellum, with a particular interest in its regional difference. At embryonic day 15, the ROR alpha mRNA was expressed at the highest level in the PC plate. The prominent expression in PCs was maintained from late embryonic stage through mature stage. At any developmental stages, no apparent regional differences in the ROR alpha mRNA expression were detected in the mediolateral and rostrocaudal axes of the cerebellum. The high expression from early developmental stages provides a molecular-anatomical basis for its important role in phenotypic differentiation of PCs. However, the even distribution in the cerebellum suggests that the unique staggerer compartments are not directly related to the loss of ROR alpha function. PMID- 9220475 TI - The pelvic floor muscles: muscle thickness in healthy and urinary-incontinent women measured by perineal ultrasonography with reference to the effect of pelvic floor training. Estrogen receptor studies. AB - Maintenance of urinary continence is multifactorial and depends mainly on detrusor control and urethral closure function. The closure forces can be categorized as permanent closure forces active at rest, and adjunctive closure forces active during physical activities. The efficiency of these forces depends on the structural components in the urethral wall, the position of the bladder neck and proximal urethra, the periurethral striated muscles, and the pelvic floor muscles. By means of pudendal blockade and simultaneous recordings of pressure and cross-sectional area in the urethra, it has been demonstrated that the striated periurethral muscles and the pelvic floor muscles are of paramount importance for the closure function. This emphasizes the importance of well functioning pelvic floor muscles to obtain continence, and probably explains the rationale for the effect of pelvic floor training in treating urinary incontinence. This study presents a review of the literature on female urinary incontinence, continence mechanisms, pelvic floor muscles, and pelvic floor training. Furthermore, a review of the literature on estrogen receptors in the pelvic floor muscles is given. Perineal ultrasonography, a method for visualization and measurement of thickness of the pelvic floor muscle, was developed and evaluated. This method was used to gain information on the thickness of the pelvic floor muscles in younger physiotherapists, healthy women, and women suffering from urinary incontinence, and to evaluate the effect of pelvic floor training. Additionally, a study of the Pelvic floor muscles was performed to assess the presence of estrogen receptors. Muscle thickness seems to decrease with age. In women over age 60 years, a significantly thinner pelvic floor muscle was found compared to younger women. The muscle increment during contraction decreased significantly with age, probably reflecting a stronger pelvic floor or a better awareness of pelvic floor function in the younger women. Incontinent women had a thinner pelvic floor muscle compared to healthy women. Hypertrophy of the muscles was demonstrated in urinary-incontinent women after pelvic floor training, and the difference in thickness of the muscles in these women before training compared to healthy women was eliminated by training. pelvic floor training reduced the use of incontinence appliances and urinary leakage both in stress and urge-incontinent women. Subjectively, 60% of the women gained a positive effect of the training. In spite of the fact that training increased muscle thickness and the increment of muscle thickness during contraction, no correlation between these parameters and subjective improvement or reduced urine loss in the pad weighing test could be demonstrated. Training may strengthen the pelvic floor without effect on the multifactorial continence mechanism in cases where urinary incontinence is caused by destruction of the urethral attachment to the surrounding tissue. No estrogen receptors were found in the nuclei of striated muscle cells in biopsies from levator ani muscles, using an immunohistochemical technique. Thus, the effect of estrogen treatment on the striated pelvic floor muscles is doubtful. A possible effect of estrogen treatment of urinary incontinence must be mediated via other structures than the pelvic floor muscles. PMID- 9220476 TI - Pressure transmission ratio: is it a reliable parameter in increased urethro vesical junction mobility? AB - Our objective was to investigate any correlation between the degree of urethro vesical junction (UVJ) mobility and the pressure transmission ratio (PTR) values. Five hundred and nineteen patients suffering from stress urinary incontinence were divided into four groups according to their degree of UVJ mobility assessed by the Q-tip test method: group 1 (N = 86), urethral axis at stress (UAS) < 30 degrees; group 2 (N = 191), UAS 31-60 degrees; group 3 (N = 214), UAS 61-90 degrees; and group 4 (N = 28), UAS > 90 degrees. A urethral pressure profile at stress was determined in the supine and standing positions, and PTR was calculated in the middle region of urethral functional length. PTR values for groups 2-4 were compared with those for group 1. In the supine position, the values for groups 3 and 4 were lower than for group 1, while in the standing position, only the values for group 3 were different. The incidence of normal PTR values (i.e., > 90%) was the same in all four groups. Overall correlation between PTR values and degree of UVJ mobility was weak (r = 0.14). We conclude that PTR values does not correlate with UVJ mobility in those patients with a Q-tip test of < 60 degrees. This correlation is inconstant when the Q-tip test was > 60 degrees. Also, 14-30% of patients in all four groups had normal standing PTR values. This may be explained by well-preserved innervation with severe alteration of the anchoring bladder neck structures. PMID- 9220477 TI - Urodynamically controlled management of spinal cord injury in children. AB - Spinal cord injuries in children are relatively uncommon. However, infants with cervical spine injury have an especially high risk of renal damage. Six patients, 4 of them tetraplegic, aged 15 months to 8 years, were primarily treated by oral anticholinergic medication and intermittent catheterization. With this concept, satisfactory results were achieved in 4 of 6 children for a mean follow-up of 17.7 months. Mean bladder capacity increased by 128% and intravesical pressure was reduced by 35%. While all patients initially presented with a detrusor leak point pressure above 40 cm H2O, in 4 patients detrusor leak point pressure could be sufficiently reduced by initial treatment. One patient required intravesical instillation of oxybutynin; in another patient sphincterotomy was performed. No patient had signs of renal damage. In summary, even in tetraplegic infants, oral anticholinergic medication and intermittent catheterization is a safe and well tolerated treatment. PMID- 9220478 TI - Efferent and afferent neuronal hypertrophy associated with micturition pathways in spontaneously hypertensive rats. AB - Elevated nerve growth factor secreted by bladder smooth muscle may be associated with noradrenergic hyperinnervation of the bladder and hyperactive voiding in spontaneously hypertensive rats (SHR) and rats with bladder outlet obstruction. The present study was undertaken to determine if changes occur in efferent and afferent pathways supplying the SHR bladder similar to those in rats with bladder outlet obstruction. Fluoro-Gold (FG) retrograde tracing studies were conducted to examine the postganglionic efferent limb (major pelvic ganglion; MPG) and sensory afferent limb (L1, L2, L6, and S1 dorsal root ganglion; DRG) of the micturition reflex pathway of the SHR and Wistar-Kyoto (WKY) normotensive rat. A significant increase in cross sectional area profiles for labeled neurons in the MPG was observed in SHRs (830.5 +/- 9.0 microns2) as compared to WKYs (736.3 +/- 16.6 microns2). Neuronal cell areas in L2 (1,010.9 +/- 18.6 microns2) and S1 (1,024.6 +/- 28.3 microns2) of SHRs were significantly larger than those of WKYs (L2, 865.3 +/- 12.6 microns2, S1, 778.3 +/- 11.2 microns2). There was an increase in number of labeled cells in L6 within SHRs over WKYs. These results provide evidence that both efferent and afferent changes in neuronal innervation of the bladder occur in SHRs. The SHR strain may represent a genetic model to study changes in micturition reflex pathways that result from alterations in neuronal morphology such as those that occur with urethral outlet obstruction. PMID- 9220479 TI - Micturition by functional magnetic stimulation in dogs: a preliminary report. AB - The effectiveness of functional magnetic stimulation (FMS) technology on bladder contraction and bladder emptying was evaluated in ten normal neurologically intact male dogs. In seven animals, FMS of the bladder was performed by using a commercially available magnetic coil (non-water-cooled) for stimulating the sacral nerves or over the suprapubic region. With sacral stimulation, the mean change in bladder pressure (Pves) was 68.0 +/- 12.96 cm H2O; with suprapubic stimulation, the mean change in Pves was 40.7 +/- 8.08 cm H2O. This change in Pves by sacral stimulation was higher than suprapubic stimulation (P < 0.01). In three additional animals, voiding was demonstrated by using a specialized water cooled magnetic coil and by stimulating the sacral nerves with an intermittent stimulation sequence. Voiding occurred in all three animals and was reproducible. In summary, FMS of the bladder has the potential to be a useful non-invasive technology for bladder emptying and bladder training in patients with neurogenic bladder. PMID- 9220480 TI - Pelviureteral inhibitory reflex and ureteropelvic excitatory reflex: role of the two reflexes in regulation of urine flow from the renal pelvis to the ureter. AB - The mechanism by which the ureteropelvic junction (UPJ) regulates the passage of urine from the renal pelvis to the ureter, and prevents urinary backflow from the the ureter to the renal pelvis, is not completely understood. The current communication studies this mechanism in 18 dogs. With the dogs under anesthesia, nephrostomy was done through which two catheters (one pressure and one balloon tipped) were introduced into the UPJ and the renal pelvis, respectively. Renal pelvis distension with a balloon filled with 1 ml of saline effected a rise of renal pelvic pressure from a mean basal pressure of 4.8 +/- 1.2 cm H2O to 6.9 +/- 2.3 cm H2O (P < 0.05). The basal UPJ pressure of 12.6 +/- 2.7 cm H2O showed no significant change with 1 ml distention of the renal pelvic balloon (P > 0.05). Renal pelvic distension with 2, 3, and 4 ml caused a significant rise of renal pelvic pressure to 8.4 +/- 2.7 (P < 0.05), 10.6 +/- 2.2 (P < 0.01), and 11.8 +/- 1.9 (P < 0.01) cm H2O, respectively, and a significant drop of UPJ pressure to 4.8 +/- 1.2, 4.7 +/- 1.1, and 4.6 +/- 1.2 cm H2O (P < 0.01), respectively. Ureteric distension with a balloon filled with 0.5 ml of saline significantly raised the ureteric pressure from a mean basal value of 4.3 +/- 1.4 cm H2O to 14.7 +/- 3.3 cm H2O (P < 0.01) and the UPJ pressure to a mean of 20.8 +/- 3.8 (P < 0.05). Ureteric distension with 1 and 1.5 ml of saline led to an elevation of ureteric and UPJ pressure which was not significantly different from that observed with distension with 0.5 ml (P > 0.05). In contrast, the UPJ showed no significant pressure change upon distension of the locally anesthetized renal pelvis or ureter, respectively. Likewise, the locally anesthetized UPJ exhibited no significant pressure response to renal pelvic or ureteric distension. The study demonstrates that urine might have to accumulate in the renal pelvis up to a certain volume and pressure so as to effect UPJ opening, which occurs at its maximum irrespective of the distending volume. UPJ opening upon renal pelvic distension postulates a reflex relationship which we call "pelviureteral inhibitory reflex." This reflex is believed to regulate the passage of urine from the renal pelvis to the ureter. Ureteric distension closes the UPJ; we call this reflex action the "ureteropelvic excitatory reflex" as it seems to prevent reflux of urine through the UPJ and thus protects the kidney. The concept that the UPJ acts as a physiologic sphincter is put forward. PMID- 9220481 TI - A clinical historical overview of pesticide health issues. PMID- 9220482 TI - Epidemiology of pesticide poisonings in the United States, with special reference to occupational cases. PMID- 9220483 TI - Pesticide exposure assessment of workers and their families. PMID- 9220484 TI - Diagnosis and treatment of organophosphate and carbamate intoxication. AB - Organophosphate and carbamate insecticides are the most common forms of pesticide poisoning. They present with a distinctive clinical syndrome that is, nevertheless, sometimes difficult to recognize. Early treatment is important, and specific antidotal therapy is available using atropine and pralidoxime. Important complications that need to be monitored during therapy may involve the central and peripheral nervous system, lungs, and heart. PMID- 9220485 TI - Basic toxicology of pesticides. AB - This chapter reviews basic aspects of the health effects and mechanisms of action of the major classes of pesticides: insecticides, herbicides, rodenticides, and fungicides. Specific pesticides in each class are described, with historical detail and depictions of chemical structure. PMID- 9220486 TI - Pesticides and cancer. AB - The authors discuss pesticide exposure in both agricultural and nonagricultural occupations, referring to a variety of studies on the cancer risks of specific pesticides. Adult and childhood cancer are addressed, and direction for future research efforts is offered. PMID- 9220487 TI - Chronic neurologic effects of pesticide overexposure. AB - Pesticide exposure in humans can have persistent effects, even in the absence of acute symptoms of intoxication or after their resolution. Drs. Keifer and Mahurin describe some of the most important examples of established pesticide neurotoxicity. PMID- 9220488 TI - Reproductive and developmental effects of occupational pesticide exposure: the epidemiologic evidence. AB - There is increasing evidence for reproductive and developmental effects of both maternal and paternal pesticide exposures. Here is a summary of the epidemiologic data, culled from studies in humans, with significant attention to Agent Orange. PMID- 9220489 TI - Skin reactions to pesticides. AB - Skin disease is the second most common type of occupational illness. This chapter reviews the important concepts in pesticide-related dermatoses, first by examining two case studies and then by describing the major fungicides, insectides, herbicides, and fumigants associated with skin disease. PMID- 9220490 TI - Monitoring the pesticide-exposed worker. AB - Cholinesterase inhibitors-the organophosphates and the carbamates-are the most acutely toxic and widely used insecticides. They also comprise the only group of pesticides for which state laws exit requiring worker monitoring. This chapter focuses on cholinesterase monitoring, with attention to available assays and testing kits. PMID- 9220491 TI - Toxicity testing of pesticides sold in the United States. PMID- 9220492 TI - State pesticide regulatory programs: themes and variations. AB - State pesticide regulation varies with region, with the amount and type of agriculture, with pesticide use, and with political conditions that are sometimes volatile and unpredictable. Dr. Arne offers an overview of state regulatory programs and their connections to the U.S. Environmental Protection Agency. PMID- 9220493 TI - Emerging issues in pesticide health studies. PMID- 9220494 TI - Appendix: general pesticide information on the Worldwide Web. PMID- 9220495 TI - Modulation of matrix metalloproteinase-7 (matrilysin) secretion in coculture of human colon carcinoma cells with fibroblasts from orthotopic and ectopic organs. AB - Matrix metalloproteinase-7 (MMP-7) is a member of the family of matrix-degrading metalloproteinases that are believed to contribute to the complex process of cancer invasion and metastasis. The secretion level of MMP-7 as assayed by immunoblot analysis was low but distinct in the culture medium of a human colon carcinoma cell line, WIDr, whereas none of the fibroblasts secreted the detectable level of MMP-7. The coculture of WiDr with various human fibroblasts from orthotopic (colon) and ectopic (thyroid, brain, lung, and skin) organs significantly stimulated the secretion of MMP-7 compared with the cultures of individual cells. Reverse transcriptase-polymerase chain reaction analysis and RNA blot analysis suggested that this enhancement occurred at a pretranslational level. The extent of the stimulation was widely varied by the fibroblasts used and was dependent on the cellular ratios and density in the coculture. There may exist a tendency that fibroblasts of orthotopic origin stimulate more extensively than do those of ectopic origin. Moreover, in the coculture of high cell density, normal fibroblasts from the ectopic organs reduced the MMP-7 secretion. The stimulation of MMP-7 secretion may be partially mediated through soluble factor(s); however, direct cell-cell interactions would be required for maximum stimulation. The enhanced MMP-7 secretion was also observed in coculture of colon fibroblasts with other colorectal carcinoma cell lines such as RCM-1 and SW837, which secreted hardly detectable levels of MMP-7 in the individual culture. These results suggest that MMP-7 secretion by colon carcinoma cells is influenced by specific interactions between the carcinoma cells and host fibroblasts. PMID- 9220496 TI - Antiproliferative effects of cyclopentenyl cytosine (NSC 375575) in human glioblastoma cells. AB - Cyclopentenyl cytosine (CPEC) exerts an antiproliferative effect against a wide variety of human and murine tumor lines, including a panel of human gliosarcoma and astrocytoma lines. This effect is produced primarily by the 5'-triphosphate metabolite CPEC-TP, an inhibitor of cytidine-5'-triphosphate (CTP) synthase (EC 6.3.4.2). Because previous studies with human glioma cell lines utilized cells in long-term tissue culture, we have undertaken to determine whether the activity of CPEC in such model systems is also demonstrable in freshly excised human glioblastoma cells. Glioma cells obtained at surgery and in log phase growth were exposed to the drug at levels ranging from 0.01 to 1 microM for 24 h, and CPEC-TP and CTP levels were determined by HPLC. Dose-dependent accumulation of CPEC-TP was accompanied by a concomitant decrease in CTP pools, with 50% depletion of the latter being achieved at a CPEC level of ca. 0.1 microM. Human glioma cell proliferation was inhibited 50% by 24-h exposure to 0.07 microM CPEC. Postexposure decay of CPEC-TP was slow, with a half-time of 30 h. DNA cytometry showed a dose-dependent shift in cell cycle distribution, with an accumulation of cells in S-phase. The pharmacological effects of CPEC on freshly excised glioblastoma cells are quantitatively similar to those seen in a range of established tissue culture lines, including human glioma, colon carcinoma, and MOLT-4 lymphoblasts, supporting the recommendation that the drug may be advantageous for the treatment of human glioblastoma. PMID- 9220497 TI - Variations in the expression of the adenomatous polyposis coli (APC) tumor suppressor gene in human cancer cell lines of different tissue origin. AB - The adenomatous polyposis coli (APC) tumor suppressor gene APC is mutated in familial adenomatous polyposis and in most sporadic colorectal tumors. Through its interaction with beta-catenin the APC protein may play a role in a signal transduction pathway regulating cell proliferation. Despite the fact that APC is ubiquitously expressed, mutations leading to truncated proteins are restricted to tumors of the digestive system. To determine further alterations not resulting in protein truncation, but possibly influencing the signaling, we compared the relative expression level of the APC protein and transcripts in 24 human colorectal cancer cell lines and in additional 17 lines of noncolorectal tissue origins, which have not previously been studied. By Western analysis, the highest levels of full-length APC protein were found in a subset of neuroblastoma and retinoblastoma cell lines. In contrast, in five noncolorectal lines it was not detectable. Truncated APC was exclusively found in 18 of the 24 colorectal cancer cell lines, but was never detected in any cell line derived from other tissues. In most colorectal cancer cell lines the protein level of full-length or mutated APC was reduced. By the more sensitive immunoprecipitation analysis, weak expression of full-length APC could be shown even in those noncolorectal cancer cell lines where it was not detectable by Western blotting. In addition, APC transcript expression was found in all cell lines, the level in colorectal cancer cell lines being reduced. PMID- 9220498 TI - Ultrastructural and phenotypic characterization of CABA I, a new human ovarian cancer cell line. AB - We have established an ovarian cancer cell line (CABA I) from ascitic fluid obtained from a patient with papillary adenocarcinoma of the ovary prior to drug treatment. The epithelial origin of the cell line was confirmed by morphology and by immunofluorescence analysis using anticytokeratin antibodies. Ultrastructural analysis revealed a very irregular membrane surface and a clear cytoplasm rich in electron-lucent vesicles. CABA I cells grow rapidly in culture (doubling time 18 h) in an anchorage-independent manner. Exogenously added beta-estradiol and epidermal growth factor (EGF) treatments did not influence cell growth rate. FACS analysis to determine the phenotypic profile of tumor-associated antigen, membrane receptor, and adhesion molecule expression indicated that the cell line was positive for different members of the c-erbB family, for alpha 6 and beta 1 integrin receptors, and intensively positive for HLA class I antigens and the folate receptor. Molecular characterization revealed no mutations for c-myc and c k-ras genes, but did detect an exon 5 mutation in the p53 gene. CABA I cells grew poorly as heterotransplants in nude mice, and tumors showed long latency periods. Because early (15-20) and late (55-60) passage cells maintain the same growth and phenotypic characteristics, the CABA I cell line might provide a good in vitro model system to investigate the cellular and molecular events involved in ovarian carcinogenesis. PMID- 9220499 TI - Thymidylate synthase as a target for growth inhibition in methotrexate-sensitive and -resistant human head and neck cancer and leukemia cell lines. AB - Thymidylate synthase (TS) inhibitor effects on growth of human head and neck squamous cell carcinoma (HNSCC) cell lines and CCRF-CEM human leukemia cells and sublines with acquired methotrexate (2,4-diamino-10-methylpteroylglutamic acid) (MTX) resistance were studied. During 120-h treatment, HNSCC cell lines A253 and FaDu are equally sensitive to MTX, whereas the polyglutamylatable TS inhibitors ZD1694 and BW1843U89 are 5- to 35-fold more potent than MTX and the lipophilic AG331 is approximately 10(2)-fold less potent than MTX. A253 is intrinsically resistant to intermittent (24 h) MTX and BW1843U89 exposure (higher EC50 values and shallower slopes of concentration-response curves relative to FaDu); AG331 and ZD1694 largely overcome this intrinsic resistance to intermittent exposure. Thymidine (TdR) protects against growth inhibition by these inhibitors, confirming that TS is their target in HNSCC; at high AG331 levels, TdR only partially protects, implying that a second site of action exists. Growth inhibition of HNSCC by ZD1694 and BW1843U89 is protected by leucovorin (LV) at > or = 10(-7) and > 10(-3) M, respectively; 10(-4) M LV cannot protect HNSCC cells against AG331. Results similar to protection studies are obtained if LV addition is delayed < or = 24 h after ZD1694 or BW1843U89 exposure. CCRF-CEM sublines with acquired MTX resistance resulting from dihydrofolate reductase (DHFR) overexpression, defective MTX transport, or defective MTX polyglutamylation retain full sensitivity to AG331. Cells with defective MTX transport are highly cross-resistant to ZD1694 and BW1843U89, implicating the reduced folate/MTX carrier in their transport. Minor cross-resistance of the DHFR overexpressing line to ZD1694 and BW1843U89 is observed. A subline with highly defective MTX polyglutamylation is cross-resistant to 120-h exposure to ZD1694, but not to BW1843U89, suggesting a profound contribution of polyglutamylation to the mechanism of action of ZD1694. PMID- 9220500 TI - O6-Alkylguanine-DNA alkyltransferase in normal colon tissue and colon cancer. AB - O6-Alkylguanine-DNA alkyltransferase (AGT) is a DNA repair protein that reverses alkylation damage produced by chloroethylnitrosoureas and is a major determinant of cellular resistance to adjuvant chemotherapy with these drugs. AGT activity was measured in 119 samples from 69 patients, including normal, tumor, and diseased tissue, and 42 patients in which both normal and tumor tissue were assayed. The activity varied among individuals, but there was no statistically significant difference in average AGT activity among tumor, normal, and diseased tissue, or between men and women, or between young and old patients (< 70 or > 70 years). Few (3/49) tumor samples showed an absence of AGT activity (Mer- phenotype). The results indicate that nearly all colon cancers have significant AGT activity, and adjuvant chloroethylnitrosoureas chemotherapy must be modified, perhaps by the use of AGT biochemical modulators, to overcome this natural drug resistance. PMID- 9220501 TI - A prospective study of the usefulness of clinical and laboratory parameters for predicting percentage of dehydration in children. AB - To evaluate the relative utility of clinical and laboratory parameters of dehydration in children for predicting the magnitude of percent less of body weight (PLBW), we studied 97 children who required intravenous fluids for acute dehydration. After a complete history and physical examination, the managing physician made a clinical estimation of dehydration for each child, based on a standard clinical scale. Serum electrolytes were obtained in all children prior to intravenous hydration therapy. PLBW was calculated after recovery from acute dehydration by comparing the weight on presentation to the emergency department with the weight measured at a follow-up visit when the child was judged well. Children were classified according to PLBW into three groups which reflect the categories in a standard clinical scale: mild = PLBW < or = 5 (n = 50), moderate = PLBW 6-10 (n = 30), and severe = PLBW > 10 (n = 17). The physician's clinical estimate of dehydration compared to PLBW had a sensitivity of 74% (95% confidence interval (CI): 60-85) for mild dehydration, 33% (95% CI: 17-53) for moderate dehydration, and 70% (95% CI: 44-89) for severe dehydration. There was a significant difference in the mean serum bicarbonate concentrations (HCO3) between the PLBW groups (P < 0.01). The sensitivity of the HCO3 < 17 mEq/L in predicting PLBW was 77% (95% CI: 58-90) for PLBW 6-10, and 94% (95% CI: 71-100) for PLBW > 10. The combination of the clinical scale and the serum bicarbonate identified all 17 children with PLBW > 10 and 90% (27 of 30) children with PLBW 6 10. Our data suggest that physicians should not rely solely on clinical assessment to rule out severe dehydration in children, and that obtaining a serum bicarbonate may improve the accuracy of predicting serious dehydration. PMID- 9220502 TI - The significance of white-centered retinal hemorrhages in the shaken baby syndrome. AB - Retinal hemorrhages in healthy children with or without a history of associated trauma are a strong indicator of child abuse. This report describes six cases of battered infants who presented with white-centered retinal hemorrhages. We discuss potential mechanisms for the presence of white-centered retinal hemorrhages in battered children. PMID- 9220503 TI - Tibial length following intraosseous infusion: a prospective, radiographic analysis. AB - Intraosseous infusion is a well accepted means of obtaining emergency intravascular access in children. Despite the low incidence of serious complications from intraosseous infusions, the potential exists for growth plate injury and subsequent growth disturbance following intraosseous infusion. We conducted a prospective, blinded observational study of 10 subjects to evaluate tibial length discrepancy radiographically one year or more following intraosseous infusion. We found no significant difference in mean tibial length between the legs that had intraosseous infusions and the opposite legs, which served as controls. We conclude that intraosseous infusion does not appear to produce subsequent leg length discrepancy one year after infusion. PMID- 9220504 TI - The value of end-tidal CO2 monitoring when comparing three methods of conscious sedation for children undergoing painful procedures in the emergency department. AB - BACKGROUND: Many studies have evaluated conscious sedation regimens commonly used in pediatric patients. Recent advances in capnography equipment now enable physicians to assess respiratory parameters, specifically end-tidal CO2 (et-CO2), more accurately in spontaneously breathing sedated children than was possible in the earlier studies. This study was designed to: 1) compare the safety and efficacy of intravenous fentanyl, intravenous fentanyl combined with midazolam, and intramuscular meperidine-promethazine-chlorpromazine (MPC) compound when used for painful emergency department (ED) procedures: and 2) to determine whether the addition of et-CO2 monitoring enabled earlier identification of respiratory depression in this population. METHODS: Forty-two children requiring analgesia and sedation for painful ED procedures were randomly assigned to receive either fentanyl, fentanyl-midazolam, or MPC compound. Vital signs, oxygen saturation, and et-CO2 were monitored continuously. Pain, anxiety, and sedation scores were recorded every five minutes. RESULTS: Respiratory depression (O2 saturation < or = 90% for over the minute or any et-CO2 > or = 50) occurred in 20% of fentanyl, 23% of fentanyl-midazolam, and 11% of MPC patients (P = NS). Of those patients manifesting respiratory depression, 6/8 were detected by increased et-CO2 only. MPC patients required significantly longer periods of time to meet discharge criteria than fentanyl and fentanyl-midazolam patients (P < 0.05). No differences were noted in peak pain, anxiety, or sedation scores. CONCLUSIONS: Fentanyl, fentanyl-midazolam, and MPC produced a high incidence of subclinical respiratory depression. End-tidal CO2 monitoring provided an earlier indication of respiratory depression than pulse oximetry and respiratory rate alone. MPC administration resulted in a significantly delayed discharge from the ED. PMID- 9220505 TI - Utility of toxicology screening in a pediatric emergency department. AB - OBJECTIVE: To determine the types of patients who undergo toxicology screen testing (TS) and the clinical utility of the test in a pediatric emergency department. DESIGN: Retrospective chart review. SETTING: Urban pediatric emergency department. PATIENTS OR PARTICIPANTS: All patients, n = 338, less than 18 years of age who had a TS sent from the Kosair Children's Hospital Emergency Department between 1/1/91 and 12/31/91. RESULTS: Three hundred and thirty-eight charts were available for review from 344 patients who had TS testing. Seventy eight patients (23%) were less than 12 years old; 164 patients (49%) were female. Forty-four patients were tested by serum TS only; 195 patients by serum plus urine TS; 94 patients by urine TS; four patients by serum, urine, and gastric aspirate TS, and one patient by urine and gastric aspirate TS. Chief complaints of patients who had TS sent were as follows: ingestion (211), abnormal behavior (56), seizures (30), trauma (18), syncope/tingling (7), depression/suicide (6), chest pain/palpitations (3), headaches (3), and other (4). While 195 patients (57%) had positive TS for at least one item, only 22 patients (7%) had a positive TS for an unexpected item, including seven patients with ingestions, eight with abnormal behavior, four with seizures, two with syncope, and one with trauma. Only three patients with unexpected positive TS had a change in medical management as a result of the TS findings. All three of these patients had abnormal physical examinations. CONCLUSION: A minority of patients have unexpected TS results. TS results rarely necessitate a change in medical management. Emergency physicians should reevaluate indications for TS testing in pediatric patients. PMID- 9220506 TI - Skull fractures in infants and predictors of associated intracranial injury. AB - BACKGROUND: Emergency department (ED) management of skull fractures in children remains controversial. Because infants incurring head trauma have a high incidence of skull fracture, we chose to describe fractures in this subset of patients and to determine if there are clinical predictors of associated intracranial injury (ICI) that may have utility in developing more efficient management schemes in these patients. METHODS: A retrospective medical record review was conducted on all awake patients < 13 months of age with an acute skull fracture from non-birth trauma, presenting to the ED of a university-affiliated children's hospital during a three-year period. Clinical and radiographic data extracted were used to describe skull fractures in these patients. The ability of various characteristics to determine the presence of ICI was assessed by calculating sensitivity, specificity, positive predictive value, and negative predictive value for each. RESULTS: The predominant mechanism of injury for the 102 infants was falls (91%). Suspicion of abuse was found in only one case. The parietal bone was fractured in 87 infants, and 34 had nonparietal fractures. The most prevalent fracture type was linear (92 infants), and 31 had > 1 cranial bone fractured. CT scans obtained on 32 infants (CT group) revealed 21 ICIs in 15 patients. Two with temporoparietal fractures required emergent evacuation of epidural blood. In the CT group, seven of the 15 (47%) with ICI (ICI group) were lethargic compared to two of the 17 (12%) without ICI (No ICI group) (P = 0.035). Five (33%) in the ICI group had temporal bone fractures compared to 0 in the No ICI group (P = 0.015). The presence of any sign or symptom had a sensitivity and negative predictive value of 100%, but only a specificity of 35%. The presence of lethargy had a positive predictive value of 78%. The presence of temporal and frontal bone fractures had positive predictive values of 100 and 75%, respectively. CONCLUSION: This study reports a high prevalence of fracture characteristics often associated with inflicted injury in other studies when virtually all injuries in our sample were accidental. Several clinical characteristics were demonstrated to be potentially useful in predicting ICI associated with skull fracture; however, prospective study is recommended to validate these findings prior to clinical application. PMID- 9220507 TI - Diagnostic use of anion and osmolal gaps in pediatric emergency medicine. AB - Analyzing anion and osmolal gaps can help in the diagnosis and management of clinically elusive cases. This article presents two such cases, and reviews the clinical use of gaps. PMID- 9220508 TI - Pneumomediastinum following penetrating oral trauma. AB - Pneumomediastinum can result from a puncture wound or laceration to the hypopharynx. This is a case report of an 18-month-old child who fell with a pen in his mouth. Initial physical examination was unremarkable, but the child developed neck swelling, fever, and irritability over the next 12 hours. Repeat examination revealed marked pneumomediastinum and subcutaneous emphysema. The pathophysiology and treatment of pneumomediastinum are reviewed. PMID- 9220509 TI - Dystonic reaction associated with dextromethorphan ingestion in a toddler. AB - INTRODUCTION: Accidental ingestions of cough and cold preparations containing dextromethorphan (DM) are common in the toddler age group and rarely have serious consequences. Even large intentional overdoses by adults seldom lead to serious morbidity. There have been no previous reports of an extrapyramidal reaction due to a DM ingestion. CASE REPORT: We report a 30-month-old girl who ingested approximately 38 mg/kg dextromethorphan. She presented with opisthotonus, ataxia, and bidirectional nystagmus. There was no change in her status with the administration of naloxone. The child was given diphenhydramine with clearing of her opisthotonus but persistence of her ataxia and nystagmus. DISCUSSION: A moderate ingestion of dextromethorphan in a toddler resulted in extrapyramidal symptoms with opisthotonus that responded to diphenhydramine. Dextromethorphan is known to have complex CNS effects and, in sufficient doses, may have dopamine receptor blocking activity resulting in this dystonic reaction. PMID- 9220510 TI - Aseptic meningitis from trimethoprim-sulfamethoxazole in an HIV-infected adolescent. AB - Adolescents infected with the human immunodeficiency virus (HIV) often confront the clinician with difficult medical problems. Besides the host of opportunistic infections, which can affect these patients, side effects from medications can be frequent and, at times, life-threatening. We report a case of aseptic meningitis secondary to trimethoprim-sulfamethoxazole therapy for prophylaxis against Pneumocystis carinii in an HIV-infected adolescent. PMID- 9220511 TI - Roller coasters: let the rider beware. AB - PURPOSE: To describe a pediatric patient with a severe abdominal injury following a roller coaster crash, and to review the relevant literature of lap belt injuries and roller coaster safety regulations. METHODS: Case report. RESULTS: A seven-year-old girl sitting in the front seat of a two-person roller coaster car was injured when it crashed into the stopped car in front. The patient's injuries, including a partial hepatic amputation, were due to the combined forces of both passengers applied against her lap belt. CONCLUSIONS: Roller coaster restraint systems do not have the same federal or state oversights as motor vehicles and can result in life-threatening injuries. PMID- 9220512 TI - Uvulitis caused by anaerobic bacteria. AB - OBJECTIVE: I present two children with bacteremic uvulitis due to anaerobic bacteria. RESULTS: Fusobacterium nucleatum was recovered from the blood, and Haemophilus influenzae type b was recovered from a surface uvular culture of one patient. beta-Lactamase-producing Prevotella intermedia was isolated for the blood of the other patient. Both patients responded to parenteral, followed by oral, antimicrobial therapy. CONCLUSIONS: These findings illustrate the need to send blood cultures for both aerobic and anaerobic bacteria in patients with uvulitis. PMID- 9220513 TI - Posttraumatic distress in children and families after intubation. AB - BACKGROUND: Posttraumatic stress disorder symptoms are seen in children who have experienced significant trauma. Respiratory arrest with subsequent intubation can be associated with terror, helplessness, and the threat of death. METHODS: Three case reports are presented where emergency intubation was followed by symptoms of psychologic distress in the intubated child and his or her family members. RESULTS: Although the medical literature documents posttraumatic distress symptoms after other medical procedures, this is the first account of symptoms following intubation. Children and other family members were found to have symptoms of reexperiencing the traumatic event, avoidance of thoughts or feelings related to the intubation, and hyperarousal. Issues around diagnosis and treatment are discussed. CONCLUSIONS: Children with a history of emergency intubation should be evaluated for possible posttraumatic stress disorder symptoms. PMID- 9220514 TI - Complications of Fleet enema administration and suggested guidelines for use in the pediatric emergency department. AB - INTRODUCTION: Hypertonic sodium phosphate enema solutions are commonly used for the treatment of acute constipation in the pediatric emergency department. The potential for severe metabolic derangement and death in children with gastrointestinal and/or renal abnormalities and these reported as normal has been documented in the literature. OBJECTIVE: To develop guidelines for the safe and effective use of hypertonic sodium phosphate enema solutions such as Fleet enema, in the pediatric emergency department setting. RESOURCES: A comprehensive review of the literature was conducted using the MEDLINE database for the time period 1966 to September 1995. Resources used within our institution, the Hospital for Sick Children, Toronto, Ontario, included Staff of the Emergency and General Surgery Departments. SUMMARY: The guidelines provided will promote safe use of hypertonic sodium phosphate enema solutions for the treatment of acute constipation in children presenting to the emergency department. PMID- 9220516 TI - Pediatric emergency medicine: legal briefs. PMID- 9220517 TI - Quality of care. PMID- 9220515 TI - Swollen, painful scrotum after basketball injury. PMID- 9220518 TI - Case records of the LeBonheur Children's Medical Center: a 17-month-old girl with abdominal distension and portal vein gas. PMID- 9220519 TI - Osteogenesis imperfecta (OI), may be mistaken for child abuse. PMID- 9220520 TI - Serum eosinophil cationic protein as a predictor of wheezing after bronchiolitis. AB - We have evaluated the role of eosinophil cationic protein (ECP) concentrations in serum in predicting wheezing after bronchiolitis, during infancy and early childhood. A prospective study at a university hospital serving all pediatric patients in a defined area was designed. Serum ECP concentrations were measured in 92 infants under the age of 2 years on admission for acute bronchiolitis, and 6 and 16 weeks after hospitalization. Nebulized anti-inflammatory therapy was initiated during hospitalization: 32 patients received cromolyn sodium and 32 patients received budesonide for 16 weeks; 30 control patients received no maintenance therapy. The numbers of subsequent physician-diagnosed wheezing episodes and hospital admissions for obstructive airway disease were recorded during 16 weeks of follow-up. At entry, 14 of 92 (15%) children had high (> or = 16 micrograms/L) levels of ECP in their serum. During the 16-week follow-up period, this group of patients had significantly more physician-diagnosed episodes of wheezing (86% vs. 43%, P < 0.01) and hospital admissions for wheezing (64% vs. 19%, P = 0.001) than those with serum levels of ECP < 16 micrograms/L. The number of patients with serum ECP > or = 8 micrograms/L was 25 (27%); 76% of this group developed physician-diagnosed wheezing (P < 0.01), and 48% had hospital admissions for wheezing (P < 0.01). Serum ECP levels decreased significantly with respect to time after bronchiolitis and did not differ among the three intervention groups. We conclude that a high serum ECP concentration during the acute phase of bronchiolitis is a specific but insensitive predictor of wheezing after bronchiolitis. PMID- 9220521 TI - Differential inhibitory effect of regular inhaled corticosteroid on airway responsiveness to adenosine 5' monophosphate, methacholine, and bradykinin in symptomatic children with recurrent wheeze. AB - Indirect tests of bronchial responsiveness to agents such as adenosine 5' monophosphate (AMP) or bradykinin might be more specific markers of a therapeutic responses to anti-inflammatory treatment than a test of direct responsiveness to agents such as methacholine. In children selected from the community on the basis of mildly symptomatic wheeze, we compared in a randomized, double-blind study design the effect of 400 micrograms/day of beclomethasone dipropionate (BDP) or placebo on three separate ways of provoking bronchial responsiveness, using methacholine, bradykinin, and AMP as the provoking agents. Following pretreatment bronchial challenges, 29 children received paired monthly methacholine and AMP challenges for 3 months, while for the same period another 33 children received paired monthly methacholine and bradykinin challenges. Compared with placebo treated subjects, FEV1 increased significantly in the children receiving BDP. This improvement was observed in those randomized to either the AMP challenge or the bradykinin challenge. In children challenged with AMP, the PD20 AMP increased significantly after 1 month and 2 months of BDP therapy when compared with placebo, while under similar conditions the PD20 methacholine was not significantly affected. In children challenged with bradykinin, BDP therapy did not significantly alter either the PD20 bradykinin or PD20 methacholine. We conclude that a bronchial challenge with AMP appears to be a more sensitive predictor of response to anti-inflammatory treatment than either methacholine or bradykinin. PMID- 9220522 TI - Effect of capsaicin on airway responsiveness to hypertonic saline challenge in asthmatic and non-asthmatic children. AB - Recurrent cough and asthma are common problems in children. In the evaluation of children with recurrent cough, the sequential measurements of airway responsiveness (AR) and capsaicin cough receptor sensitivity may be useful. However, the effect of capsaicin on AR induced by an indirect stimulus such as hypertonic saline (HS) is not known. Current evidence suggests that a common pathway is involved in both capsaicin and HS challenges. This study was designed to determine whether inhalation of capsaicin for the cough receptor sensitivity test before HS challenge will alter AR of asthmatic and non-asthmatic children to that challenge. Twenty-one children (12 asthmatics, 9 non-asthmatics; mean age, 11.3 years) performed the HS challenge alone or 2 min after capsaicin inhalation on 2 different days in random order. The end point of the capsaicin inhalation was when > or = 5 coughs were stimulated from a single inhalation. The power of the study was > 90% at a significance level of 0.05. Capsaicin inhalation prior to HS challenge did not alter the AR of normal children. In the asthmatic group, the PD15 (provocation dose causing a fall in forced expiratory volume in 1 s of > or = 15% from the baseline) without prior inhalation of capsaicin (mean, 2.44 +/- SEM 1.21 ml) was not significantly different from that when HS challenge was performed after capsaicin inhalation (mean, 2.19 +/- SEM 0.83 ml). The mean of the difference in log PD15 of the HS challenge with and without capsaicin was 0.02 (95% CI, -0.16, 0.12), i.e. within the equivalence range of the HS challenge in children with asthma. We conclude that in normal and asthmatic children, capsaicin inhalation does not alter AR to HS; consequently the capsaicin cough sensitivity test can be performed validly before an HS challenge. PMID- 9220523 TI - Urinary cotinine and parent history (questionnaire) as indicators of passive smoking and predictors of lower respiratory illness in infants. AB - Studies of the effects of passive smoking on lower respiratory illness (LRI) have relied on questionnaires to measure exposure. We studied the association between two measures of passive smoking and the incidence of acute LRI in infants. We analyzed data from a community-based cohort study of respiratory illness during the first year of life in North Carolina. The incidence of LRI was determined by telephone calls at 2-week intervals. Environmental, demographic, and psychosocial risk factors for LRI were measured during home interviews. Tobacco smoke exposure was measured as the mean number of cigarettes smoked per day in the infant's presence. Smoke absorption by the infants was measured by the urinary cotinine/ creatinine ratio. Of the 485 infants in the study, 325 (67%) had telephone follow up and at least two home interviews. In bivariate analyses, reported tobacco smoke exposure and urinary cotinine were associated with LRI. Only the association between reported exposure and LRI remained significant after adjusting for confounders, [adjusted incidence of LRI (episodes/child-year) non exposed: 0.6; < or = 10 cigarettes/day: 0.9 (RR 1.5, 95% CI: 1.1, 2.0); > 10 cigarettes/day: 1.3 (RR 2.2, 95% CI: 1.3, 3.8)]. We conclude that infants reportedly exposed to tobacco smoke have an increased incidence of LRI. There are differences between questionnaire and biochemical measures of passive smoking. Urinary cotinine will not necessarily improve the validity of studies of the relationship of passive smoking to LRI in infants. PMID- 9220524 TI - Experience with intubated patients does not affect the accidental extubation rate in pediatric intensive care units and intensive care nurseries. AB - Accidental extubation is a potentially serious event for pediatric or neonatal patients with respiratory failure, especially in clinical settings in which personnel capable of performing reintubation may not be readily available. Thus the rate of accidental extubation in small intensive care units that operate without 24-hour in-house physician availability may be an important quality assurance indicator. The objective of this study were to determine the accidental extubation rate at a single small pediatric intensive care unit (PICU) and compare it with published reports. This study was carried out in a six-bed PICU at Washoe Medical Center in Reno, Nevada, with a relatively low level of patient acuity, as measured by PRISM score and the frequency of intubation, and without 24-hour in-house physician availability. All intubated patients admitted during the 5-year period from January 1, 1989 to December 31, 1993 were included. The primary outcome measure was the occurrence of accidental extubation. We observed only two accidental extubations in 1,749 intubated-patient-days (IPD) (0.114 accidental extubations/100 IPD [95% confidence interval 0.014-0.413 accidental extubations/ 100 IPD]). This rate of accidental extubation was compared with data in published reports from neonatal intensive care units (NICUs) and PICUs, which ranged from 0.14 accidental extubations/100 IPD to 4.36 accidental extubations/100 IPD. The dependence of the observed accidental extubation rate on unit size and institutional experience with intubated patients, as measured by the average number of intubated patients, was examined. We found no evidence that the accidental extubation rate is higher in smaller units or units with less institutional experience. Low rates can be achieved in small units with low acuity. PMID- 9220525 TI - Assessment of tidal volume over time in preterm infants using respiratory inductance plethysmography, The CHIME Study Group. Collaborative Home Infant Monitoring Evaluation. AB - Non-invasive techniques for monitoring ventilation in infants are widely used in short-term laboratory-studies but have not been evaluated in routine clinical settings. To determine whether respiratory inductance plethysmography (RIP) can provide reproducible measurements of tidal volume (VT) in premature infants over an extended period of time, we monitored respiration in eight healthy preterm infants over 4.9 +/- 1.0 hours (mean +/- SD). The algebraic sum (Sum) of rib cage (RC) and abdominal (AB) motion signals (obtained by RIP) was calculated and presented over the entire recording period as percent of an initial 5 minute calibration period. VT was simultaneously measured with a nasal mask pneumotachometer with infants in prone and supine positions during active and quiet sleep. Infants were studied in the morning (AM) and again in the afternoon (PM). Between these studies they were returned to the nursery wearing the RIP in a continuous record mode. For all patients there was a significant linear relationship between VT (in mL measured by pneumotachometer) and Sum (in % of calibration value, RIP). Neither the slope of the relationship (0.074 +/- 0.03 in AM vs. 0.071 +/- 0.02 in PM), nor its variability as measured by standard error of the estimate (SEE) (2.3 +/- 0.5 in AM vs. 2.5 +/- 0.8 in PM) changed significantly from AM to PM. The relationship between VT and Sum, as well as the variability of that relationship, was not altered by position, asynchrony of RC and AB, respiratory rate, or percent RC contribution to Sum. We conclude that RIP produces consistent measurements of respiratory effort over 5 hours in healthy preterm infants without need for recalibration and is not affected by routine care. PMID- 9220526 TI - Modification of the open circuit N2 washout technique for measurement of functional residual capacity in premature infants. AB - We compared the standard nitrogen (N2) washout technique for measuring functional residual capacity (FRC) with a modified technique that uses a helium/oxygen mixture (heliox) at different ratio instead of pure oxygen. The tests were made with a standard lung function system equipped with an ultraviolet (UV) analyzer for measurement of N2 concentrations in the expired gas. We examined models of "spontaneous breathing" and "mechanical ventilation," each with volumes of FRC in the range of a premature and a newborn lung (20-80 ml), using both techniques at different baseline inspired oxygen concentrations (FIO2). Correlations between known and measured volumes were high and identical for the two techniques (r = 0.996), and the mean error was not significantly different from zero (P = 0.111). Measurements of FRC in 6 infants gave a correlation coefficient of r = 0.989 between the two techniques; reproducibility, as measured by the coefficient of variation, was high, showing no significant differences between both techniques (P = 0.792). However, values of individual infants were different (P = 0.011), and the slope of the regression line relating measurements by the 2 techniques was 1.04, with an intercept on the y-axis at 1.46. We conclude that FRC can be measured with the modified N2 washout technique, using heliox as a washout gas. Volumes can be measured with high precision and reproducibility, even in premature infants with low lung volumes and/or high baseline FIO2. A correction factor may be necessary to equate FRC measurements made by oxygen-N2 vs. heliox N2 washouts. Hyperoxemia and hypoxemia can be avoided by admixing different flows of oxygen to a standard heliox mixture. PMID- 9220527 TI - Effects of salbutamol delivery from a metered dose inhaler versus jet nebulizer on dynamic lung mechanics in very preterm infants with chronic lung disease. AB - Treatment of chronic lung disease of prematurity requires effective aerosol delivery of different therapeutic agents. Aerosols can be generated by a metered dose inhaler (MDI) or a jet nebulizer. An MDI combined with a spacer device is easier to use and avoids undesirable effects noted in conjunction with jet nebulization. We compared the clinical effectiveness of 200 micrograms (2 puffs) salbutamol delivered from an MDI in conjunction with a valved spacer device (Aerochamber), and 600 micrograms given via jet nebulizer (PariBaby) on 2 consecutive days, the order being randomized. Thirteen spontaneously breathing very preterm infants [mean (SD) gestational age 27.2 (1.8) weeks; birth weight 0.90 (0.34) kg] were studied at a corrected age of 37 (2.3) weeks. Mean (SD) study weight was 1.83 (0.38) kg. Dynamic lung compliance and resistance were determined from measurements of flows, volumes, and transpulmonary pressures, using a pneumotachometer and a small esophageal microtransducer catheter before and 20 min after salbutamol application. Baseline values before salbutamol administration were similar on both occasions: the mean (SD) compliance was 7.7 (3.0) mL.kPa-1.kg-1 pre-MDI plus-spacer and 8.4 (3.1) pre-jet nebulizer; the resistance was 10.4 (4.0) kPa.L-1.s pre-MDI plus-spacer and 9.7 (3.4) pre-jet nebulizer. Following salbutamol, compliance did not change significantly with either MDI plus spacer or jet nebulizer. Resistance fall significantly with MDI plus spacer (mean -2.2; 99.9% CI -0.35, -4.35) and jet nebulizer (-2.4; 99% CI 0.39, -4.42). We conclude that even in small preterm infants 200 micrograms salbutamol via MDI plus spacer improves dynamic resistance as effectively as 600 micrograms via jet nebulizer and may therefore be a preferable mode of aarosol administration. PMID- 9220529 TI - Freeman Sheldon syndrome: severe upper airway obstruction requiring neonatal tracheostomy. PMID- 9220528 TI - Comparison of perfluorochemical fluids used for liquid ventilation: effect of endotracheal tube flow resistance. AB - Neonatal endotracheal tubes with small inner diameters are associated with increased resistance regardless of the medium used for assisted ventilation. During liquid ventilation (LV) reduced interfacial tension and pressure drop along the airways result in lower alveolar inflation pressure compared with gas ventilation (GV). This is possible by optimizing liquid ventilation strategies to overcome the resistive forces associated with liquid density (rho) and viscosity (mu) of these fluids. Knowledge of the effect of rho, mu, and endotracheal tube (ETT) size on resistance is essential to optimize LV strategies. To evaluate these physical properties, three perfluorochemical (PFC) fluids with a range of kinematic viscosities (FC-75 = 0.82, LiquiVent = 1.10, APF-140 = 2.90) and four different neonatal ETT tubes (Mallincrokdt Hi-Lo Jet ID 2.5, 3.0, 3.5, and 4.0 mm) were studied. Under steady-state flow, flow and pressure drop across the ETTs were measured simultaneously. Resistance was calculated by dividing pressure drop by flow, and both pressure-flow and resistance-flow relationships were plotted. Also, pressure drop and resistance were each plotted as a function of kinematic viscosity at flows of 0.01 L.s-1 for all four ETT sizes. Data demonstrated a quadratic relationship with respect to pressure drop versus flow, and a linear relationship with resistance versus flow: both were significantly correlated (R = 0.92; P < 0.01) and were inversely related to ETT size. Additionally, there was a significant correlation between pressure drop or resistance and kinematic viscosity (R = 0.99; P < 0.01). For LV in neonates these data can be used to select the optimum ETT size and PFC liquid depending OR the chosen ventilation strategy. PMID- 9220530 TI - Congenital tracheal stenosis in a patient with Down's syndrome. PMID- 9220531 TI - Diagnosis of subglottic hemangioma by chest CT. PMID- 9220532 TI - Noninvasive ventilation during percutaneous gastrostomy placement in Duchenne muscular dystrophy. AB - Noninvasive positive pressure ventilation (NPPV) is used for respiratory support in a number of diseases causing acute or chronic respiratory failure. We describe a novel use of NPPV to provide respiratory support during sedation for percutaneous placement of a gastrostomy tube in a patient with Duchenne muscular dystrophy (DMD). The patient had severe respiratory insufficiency, progressive dysphagia, and undernutrition. In addition to the case in this report, we have used NPPV to provide respiratory support to DMD patients during five other gastrointestinal endoscopies without complication. The technique is highly labor intensive and requires physicians and respiratory therapists familiar with NPPV. The primary risk associated with this technique is lack of definitive airway protection during the procedure, which must be balanced against the risks of intubation in an anesthetized patient with neuromuscular disease. The potential benefit to selected patients is substantial, such as initiation of gastrostomy tube feeding in our patient, with subsequent improvement in his quality of life and nutritional status. PMID- 9220533 TI - Atherosclerosis as a microvascular disease: impaired angiogenesis mediated by suppressed basic fibroblast growth factor expression. AB - We present evidence that hypercholesterolemia and oxidized low-density lipoprotein (ox-LDL) impair endothelial cell growth by suppressing basic fibroblast growth factor (bFGF) expression. Background studies show that diet induced hypercholesterolemia in rabbits impairs hyperplastic lumen-expanding remodeling of the carotid artery in response to a chronic flow load. Hypercholesterolemia also markedly impairs compensatory macrovascular and microvascular growth in rabbit ears with surgical restriction of arterial supply. In an in vitro model of angiogenesis, arterial explants cultured in a three dimensional collagen gel exhibited organized endothelial cell growth with formation of capillary-like microtubes (CLM). CLM growth was sensitive to inhibition by neutralizing antibodies against bFGF. With explants excised from both the aorta of hypercholesterolemic rabbits and from coronary arteries of patients with coronary arteriosclerosis, CLM growth and release of immunoassayable bFGF to the culture medium were suppressed. Growth suppression was reversed partially by exogenous bFGF. In control explants, ox-LDL produced a suppression of CLM growth that could be reversed by exogenous bFGF. In endothelial cells in culture, ox-LDL suppressed bFGF expression and DNA synthesis in a dose-dependent manner. We conclude that atherosclerosis is associated with impaired bFGF-dependent endothelial cell growth manifested by impaired adaptive growth responses of large arteries and microvessels. PMID- 9220534 TI - In vivo evidence of the critical role of cadherin-5 in murine vascular integrity. AB - Vascular endothelial cell-cell adhesion is crucial for the regulation of vascular functions and is associated with many circulatory disorders. We isolated a rat monoclonal antibody (VECD1) recognizing the mouse vascular endothelial cell adhesion molecule and found that it inhibited vascular endothelial cell-cell association. We sequenced a full-length cDNA of the antigen that was identical to mouse cadherin-5. L-cells transfected with its cDNA acquired cell-cell adhesiveness, and these transfectants reacted with VECD1 at cell-cell contact areas. We studied the role of mouse cadherin-5 in vascular functions. The addition of VECD1 antibody to a cultured vascular endothelial cell line (F-2) caused the detachment of each cell. Although normal F-2 cells formed tubular structures on Matrigel, VECD1 disturbed the tubulogenesis. VECD1 also increased the permeability through the F-2 cell layer. To clarify the in vivo function of mouse cadherin-5, we intraperitoneally injected the hybridomas producing VECD1 into adult mice. Severe venous stasis and subcutaneous hemorrhage were induced within several days after the injection, resulting in the early death of the animals. These findings are evidence of an essential role of cadherin-5 in the regulation of vascular endothelial cell-cell adhesion in vivo. PMID- 9220535 TI - Promotion of leukocyte transendothelial cell migration by chemokines derived from human biliary epithelial cells in vitro. AB - Biliary epithelial cells are the focus of inflammatory damage in several liver diseases, including allograft rejection wherein intrahepatic bile ducts are infiltrated and damaged by T cells and neutrophils. Locally secreted chemotactic cytokines (chemokines) are important signals for leukocyte recruitment to an inflammatory site and include interleukin-8 (IL-8) and monocyte chemotactic protein-1 (MCP-1), potent chemotactic agents for neutrophils and monocyte or T cells, respectively. In this study, we demonstrate that primary cultures of human biliary epithelial cells (BECs) express and secrete IL-8 and MCP-1, both of which are upregulated rapidly and markedly in response to the proinflammatory cytokines IL-1 and tumor necrosis factor-alpha. Interferon-gamma had a differential effect by reducing IL-8 secretion but stimulating MCP-1 secretion. BECs cocultured in transwell chambers below confluent monolayers of endothelial cells promoted the transendothelial migration of neutrophils, which was blocked by antibodies to CD18 or CD11b but only partially inhibited by blocking antibodies to IL-8. We conclude that human BECs produce and secrete potent, functional chemokines when stimulated by proinflammatory cytokines. The ability of BECs to secrete chemokines and thus to promote leukocyte infiltration into portal tracts seems likely to be an important cause of bile duct damage in such conditions as liver allograft rejection and may explain the involvement of intrahepatic bile ducts in a number of inflammatory liver diseases. PMID- 9220536 TI - Ehlers-Danlos syndrome type VI results from a nonsense mutation and a splice site mediated exon-skipping mutation in the lysyl hydroxylase gene. AB - We have characterized a patient with Ehlers-Danlos syndrome type VI as a compound heterozygote for the lysyl hydroxylase (LH) gene, with a pathogenetic mutation in each allele contributing to the very low levels of mRNA and LH activity in his fibroblasts. Amplification of full-length LH cDNAs resulted in normal-sized (2.9 kb) and shortened (2.8-kb) transcripts indicative of two populations of alleles. One allele contained a paternally inherited C1557 to G transition that coded for a premature stop codon (Y511X) and introduced an Nhe I restriction site in exon 14 of the LH gene. The mutation in the other allele was an exon 5 deletion that produced the shortened polymerase chain reaction transcript and generated a premature stop codon at the beginning of exon 7. Sequencing of genomic DNAs spanning exon 5 showed a mutation in the consensus donor splice site at the beginning of intron 5 (gt-->at) in both the proband and his mother. Via reverse transcriptase-polymerase chain reaction, the parents' fibroblasts showed a disproportionately lower level of each mutant allele compared to their normal alleles. This study suggests that the decreased transcription of the LH gene, which may be attributed to the presence of the nonsense mutations, accounts for the LH deficiency, and consequently, this patient's clinical phenotype of Ehlers Danlos syndrome type VI. PMID- 9220537 TI - Evidence for inflammatory and secretagogue lipids in cyst fluids from patients with autosomal dominant polycystic kidney disease. AB - Advanced autosomal dominant polycystic kidney disease (ADPKD) is characterized morphologically by massive cyst enlargement, moderate interstitial infiltration with mononuclear cells, and extensive fibrosis. In patients affected by a common genotype (PKD1), it has been suggested that the progressive decline in renal function that transpires over a highly variable time course may be due to endogenous or exogenous epigenetic factors. We have postulated that a neutral lipid, discovered in human cyst fluid and stimulating the rates of transepithelial fluid secretion and cellular proliferation of renal epithelial cells in vitro may have a potential role in cyst growth and the progressive decline of kidney function. In this study, we used thin-layer chromatography (TLC) and high-performance TLC (HPTLC) to determine whether lipid extracts of human cyst fluid stimulated monocyte chemotaxis in vitro. Monocyte chemotactic activity, determined by the transmembrane migration of murine RAW 264.7 cells, was stimulated (delta 26.0 +/- 1.5 optical density units) by a lipid fraction less polar than sphingosine but more polar by TLC and HPTLC than 1 monooleoylglycerol. A high level of secretagogue activity was detected in this fraction (delta 0.336 +/- 0.022 microliter/cm2 1 hr) and to a lesser extent (delta 0.253 +/- 0.022 microliter/cm2/hr) in a neighboring fraction that encompassed the 1-monooleoylglycerol standard. Cyst fluid with undetectable secretagogue activity had a monocyte chemotactic-activity level only 18% as great as fluids with high levels of secretagogue activity. The secretagogue and chemotactic activities in TLC-HPTLC fractions were resistant to treatment with KOH, but both were diminished by HCl, borohydride, or periodate. Rat proximal tubule cultures incubated with oleate complexed with albumin elaborated secretagogue and chemotactic activities in the conditioned medium, with TLC-HPTLC mobility characteristics similar to the biologically active cyst fluid lipids. On the basis of these studies, we conclude that human cyst fluids harbor potent secretagogue and chemotactic lipids that may have a role in determining the functional course of ADPKD. On the basis of preliminary chemical characterizations, we suggest that the secretatogue and monocyte chemotactic activities of cyst fluid may reflect the action of lipid molecules of similar structure, the source of which may be renal epithelial cells. PMID- 9220538 TI - Protection of rats against oxygen toxicity by tracheal administration of plasmid DNA: role of endogenous tumor necrosis factor. AB - Recombinant plasmid DNA alone or in conjunction with a gene delivery system has been used increasingly for in vivo gene transfer. However, little is known about the direct biological effects of plasmid DNA. In this study, we demonstrated that tracheal administration of a number of plasmid DNA protected rats against oxygen toxicity. This protection required the presence of intact plasmid DNA, was not due to endotoxin contamination, and was not shared by salmon testis DNA. The plasmid DNA-induced protection against oxygen toxicity was associated with the production of tumor necrosis factor (TNF) and the enhancement of pulmonary Mn superoxide dismutase (MnSOD), CuZnSOD, and glutathione peroxidase activities. Coadministration of plasmid DNA and anti-TNF antibody (but not nonspecific IgG) partially abolished the protective effect and reduced the pulmonary MnSOD activity, suggesting that the plasmid DNA-induced oxygen tolerance was in part mediated by the endogenous TNF and MnSOD. In view of these observations and the known immunostimulatory effects of bacterial DNA, caution should be exercised in interpreting the results of in vivo gene transfer using recombinant plasmid DNA. PMID- 9220539 TI - Modulation of globin gene expression in cultured erythroid precursors derived from normal individuals: transcriptional and posttranscriptional regulation by hemin. AB - We are interested in the genetic mechanisms whereby several classes of drugs increase fetal hemoglobin (HbF) in patients with sickle-cell anemia or beta thalassemia. Recently, we have shown (Kollia et al., Proc. Natl. Acad. Sci. U.S.A. 93: 5693, 1996) that cultured primary human adult erythroid cells (hAEC) offer a useful model for the study of transcriptional and posttranscriptional regulation of globin gene expression. We have found also that hemin markedly increases HbF levels in these cells. We report here the effect of hemin on globin gene transcription and RNA processing in hAEC. Quantitative reverse transcriptase polymerase chain reaction analysis showed that the gamma-globin message levels in the cytoplasm and nucleus were increased two-fold by hemin. In the untreated cells, only spliced gamma-transcripts were detected in the cytoplasm, indicating that only completely processed gamma-RNA is transported to the cytoplasm, whereas approximately half of the nuclear gamma-globin RNA transcripts were unspliced. After treatment with hemin, correctly spliced gamma-transcripts increased in the cytoplasm and nucleus, while the unprocessed gamma-transcripts decreased in number in the nucleus. We also studied epsilon-globin RNA transcripts; in the cytoplasm of untreated cells, only correctly processed transcripts were present, whereas the nuclear epsilon-globin RNA transcripts were unspliced. In hemin induced cells, unspliced nuclear epsilon-transcripts decreased in number. In contrast to the gamma- and epsilon-globin genes, the levels of full-length, correctly spliced beta-globin message are not affected by hemin. Nuclear run-on transcription assays confirmed the increase in the rate of transcription of gamma and epsilon-globin genes in hemin-treated versus untreated hAEC. These results indicate that hemin affects the expression of embryonic and fetal globin genes by acting both at the transcriptional and posttranscriptional levels. These results may be relevant to the action of other agents that affect the hemoglobin phenotype of human erythroid cells. PMID- 9220540 TI - Autoantigens in insulin-dependent diabetes mellitus: molecular cloning and characterization of human IA-2 beta. AB - In this study, we describe the isolation, expression, and characterization of a new member of the transmembrane protein tyrosine phosphatase family from human brain, designated IA-2 beta. The 3853-bp cDNA encodes 986 amino acids with a molecular mass of 108,044 daltons (a predicted pI value of 5.8). The intracellular domain of human IA-2 beta is 74% identical to human IA-2. Northern blot analysis showed that IA-2 beta cDNA recognized two transcripts (approximately 5.0 kb and 4.0 kb) in four of five human insulinomas, one glucagonoma, and in normal human brain, pituitary, and pancreas, but not in a variety of other normal tissues. Rabbit antiserum, raised against the intracellular domain of IA-2 beta, reacted with pancreatic islets. Treatment of in vitro-translated full-length IA-2 beta protein with trypsin converted it into a 37-kD fragment. Using recombinant human IA-2 beta, we developed a radioimmunoprecipitation assay to measure autoantibodies in the sera of patients with insulin-dependent diabetes mellitus (IDDM). Seventy-six new-onset IDDM patients were tested. Thirty-seven percent (28 of 76) of the IDDM sera-but less than 1% of the control sera (1 of 174)-reacted with IA-2 beta. The same IDDM sera tested for autoantibodies to IA-2 and glutamic acid decarboxylase (GAD65) showed that 64% (49 of 76) and 57% (43 of 76), respectively, were positive. All but two of the IA-2 beta autoantibody-positive sera also reacted with IA-2, supporting the close sequence similarity between the two molecules. Combination of any two markers, such as IA-2 beta and IA-2, or IA-2 beta and GAD65, or IA-2 and GAD65, revealed that 67%, 74%, and 87% of IDDM sera were positive for autoantibodies, respectively. Blocking of IDDM sera with recombinant IA-2, IA-2 beta, or GAD65 resulted in marked inhibition of reactivity of IDDM sera with pancreatic islet sections as measured by islet cell autoantibody immunofluorescence. This result suggests that these three autoantigens are the major targets of islet-cell autoantibody reactivity. PMID- 9220541 TI - Bronchial and alveolar allergen-induced anaphylaxis and the stimulation of bronchial mucociliary clearance in ragweed-sensitized dogs. AB - In allergic airways disease, we hypothesized that an acute allergen inhalation activates cells in the bronchial and alveolar regions of the lungs to initiate cardiopulmonary anaphylactic responses that include the stimulation of bronchial mucociliary clearance. Seven beagles were neonatally sensitized to ragweed allergen, and four were sham-sensitized. Adult dogs were anesthetized with propofol and etomidate. Bronchial retention of radiotagged particles deposited in the lungs was monitored with a gamma camera. Then 0.4-1.8 micrograms of ragweed allergen was deposited either proximally or peripherally in the lungs while achieving a similar total mass deposited. Both proximal and peripheral allergen deposition elicited cardiopulmonary responses characteristic of anaphylaxis. Following proximal allergen deposition, the mean bronchial mucuciliary clearance at 60 min increased from 27.5% +/- 4.9% to 59.9% +/- 3.3% (p < .01), and following peripheral deposition it increased from 5.9% +/- 3.1% to 52.9% +/- 7.2% (p < .01). No allergen-induced suppression of bronchial mucociliary clearance was detected within the 140-min postexposure period. No changes in cardiopulmonary responses or bronchial mucociliary clearance in the unsensitized dogs could be ascribed to the inhalation of allergen. Both the bronchi and alveoli are target sites for the initiation of allergen-induced respiratory and cardiovascular anaphylactic responses and the stimulation of bronchial mucociliary clearance. PMID- 9220542 TI - Self-management within a classroom token economy for students with learning disabilities. AB - Students with disabilities who are served in restrictive educational settings often display inappropriate behavior that serves to preclude their integration into the mainstream. One approach to managing difficult behavior is a levels system (Smith & Farrell, 1993), which typically consists of a hierarchy of levels in which students must meet increasingly demanding standards of behavior before advancing through the hierarchy. In the present study, two middle-school students with learning disabilities participated in a classroom-wide token economy based on a levels system. The levels system, which was used in a self-contained classroom, targeted the acquisition and maintenance of academic skills and social behaviors with the goal of integrating these students into an inclusive classroom. The two participants showed little or no progress within the levels system because of a very high rate of inappropriate verbalizations. Therefore, a self-management system that involved training on the accuracy of self-recording these verbalizations was added to the levels system for these students. In addition, the investigator discussed with these students the consequences of inappropriate behavior and socially appropriate behavioral alternatives. A multiple-baseline-across-subjects experimental design revealed that the intervention resulted in a substantive reduction in inappropriate verbalizations, as well as greater progress through the levels system. Implications of these findings for the use of self-recording within a token economy, the importance of students' accuracy of self-recording, and methodological issues are discussed. PMID- 9220543 TI - Psychopathology and mental retardation: an Italian epidemiological study using the PIMRA. AB - The incidence of psychopathology was studied in 176 adult patients with mental retardation through administration of the Psychopathology Instrument for Mentally Retarded Adults (PIMRA). Prevalence of answers relating to anxiety, tendency to live apart and body complaints in the group with mild mental retardation was documented. For people with severe intellectual impairments, mood disturbances with inconsistent behaviours was most prevalent. Implications of the findings are discussed. PMID- 9220544 TI - A behavioral diagnostic paradigm for integrating behavior-analytic and psychopharmacological interventions for people with a dual diagnosis. AB - Aberrant behaviors exhibited by people with developmental disabilities have been well documented. Often, psychotropic medications, especially neuroleptics, have been used to control behaviors such as self-injury, physical aggression, property destruction, and hyperactivity. Serious side effects of these medications have occurred, resulting in litigation and regulation of their use by courts, surveyors, and accrediting bodies. Rules and regulations have been developed requiring that behaviors/symptoms necessitating that medication usage be clearly delineated, that behavior programs be developed and implemented to reduce need, and that the interdisciplinary team approach be used to monitor effectiveness of interventions. Currently, little guidance exists on how behavioral and psychopharmacological interventions should be applied or combined. This paper presents a paradigm for integrating behavior-analytic and psychopharmacological treatment interventions in the treatment of persons with developmental disabilities that meets applicable standards. Our model is consistent with the least restrictive, yet effective treatment philosophy. Implications for research and treatment are presented. PMID- 9220545 TI - Comparing methods for maintaining the safety of a child with pica. AB - Pica, a potentially life-threatening behavior problem exhibited among persons with mental retardation is sometimes addressed by methods such as application of restraints to reduce or eliminate associated risks (Rojahn, Schroeder, & Mulick, 1980). However, restraints may be associated with decreases in social interaction and negative impact on quality of life. We evaluated two methods (restraint vs. no restraint) for maintaining the safety of a client with pica on three dimensions: (a) level of pica, (b) therapist effort, and (c) impact on quality of life. Both methods prevented pica, however, the no restraint condition required less therapist effort and had less negative impact on quality of life. All three dimensions were included in a clinical decision-making model to determine the least restrictive, safe level of restraint for a 4-year-old girl while assessment and treatment procedures were conducted. The clinical utility of this multifactor decision-making model is discussed. PMID- 9220546 TI - Validity of the mania subscale of the Diagnostic Assessment for the Severely Handicapped-II (DASH-II). AB - We attempted to establish the internal consistency and validity of the Diagnostic Assessment for the Severely Handicapped-II (DASH-II) to screen for the presence of mania (bipolar disorder) in severely and profoundly mentally retarded adults, Subjects included 22 individuals residing in a large developmental center in Central Louisiana. The Mania subscale of the DASH-II was internally consistent. Additionally, the DASH-II could be used to accurately classify manic and control individuals. Specific items on the subscale were examined to identify those items most associated with a diagnosis of mania. PMID- 9220548 TI - Melanoma and melanoma in situ: build a better diagnosis through architecture. AB - Conventional microscopy has remained the gold standard for melanoma diagnosis for several decades, and a diagnosis of melanoma is optimally based on a summation of microscopic features (criteria) that are evaluated as objectively as possible by an experienced histopathologist. Most pathologists and dermatopathologists assess multiple criteria before arriving at a diagnosis of melanoma, but the diagnosis remains somewhat subjective as different interpreters employ similar criteria but assemble them in very different ways. Due to the subjective aspects of the microscopic diagnosis of melanoma, considerable interobserver variability exists, even among expert diagnosticians. This article includes a brief analysis of the reproducibility of a diagnosis of melanoma with a comparison of architectural and cytological criteria. There is evidence to suggest that architectural attributes hold greater reproducibility over cytological features in the diagnosis of melanocytic neoplasms. If architectural criteria outperform cytological criteria in terms of reproducibility, then architectural features should probably be given preference over cytopathological aberrations in daily diagnosis. The author forwards four steps that can be used in the evaluation of any melanocytic neoplasm as well as an approach to melanoma diagnosis in which architectural features are emphasized. PMID- 9220547 TI - Cutaneous malignant melanoma: classification and clinical diagnosis. AB - Cutaneous malignant melanoma (MM) is a treacherous disease which carries high mortality rates. However, when diagnosed early it is wholly curable. The incidence of MM is rising steadily. The most important clinical signs include the appearance of a newly acquired pigmented lesion or change in a preexisting one. Melanoma has been classified into subtypes which include melanoma in situ, lentigo maligna melanoma, nodular melanoma, acral lentiginous melanoma, desmoplastic melanoma, superficial spreading melanoma, and mucosal melanomas. Although these overlap, there are characteristic clinical features of each that are generally recognizable. Evaluation of pigmented lesions requires correlation of clinical findings with risk factors, family history and histology. A representative skin biopsy should be performed on any lesion suspected of being MM, even if the possibility is remote. PMID- 9220549 TI - Biopsy technique for pigmented lesions. AB - The biopsy technique that should be used when sampling a pigmented lesion may not always be readily apparent. The final arbiter is whether the specimen that will be generated will be representative of the entire process so that an accurate and complete diagnosis will be able to be rendered. In some cases, melanoma may not be clinically suspected so that it is essential that any biopsy that is performed will detect these lesions. PMID- 9220550 TI - Staging and prognosis of melanoma. AB - Prognosis and survival for patients diagnosed with melanoma depends on a number of interrelated factors, including histological, clinical, immunologic, and surgical parameters. Tumor thickness and depth of invasion is the most important prognostic factor, and helps to guide treatment and management plans. Besides tumor thickness, other histological criteria include melanoma growth phase (nodal v. vertical), host response, angiolymphatic invasion, mitotic rate, regression, satellitosis, and neurotropism. Important clinical prognostic factors include anatomic location of the melanoma and the patient's age and sex. Surgical factors to be examined involve excisional margin size, and elective lymph node dissection. Finally, the staging of patients with melanomas (nodal or visceral metastases) helps to define survival. PMID- 9220551 TI - Amelanotic malignant melanoma. AB - Amelanotic malignant melanoma (AMM) often defies clinical diagnosis because of its wide range of clinical appearances and lack of pigmentation. Biopsy of AMM typically yields the correct diagnosis, although the histological findings, especially in metastatic lesions, occasionally may be confused with other malignancies. In cases histologically challenging, immunohistochemical techniques frequently provide diagnostic information. Multiple mechanisms to explain amelanosis have been suggested, all resulting in a single, common amelanotic phenotype. Appropriate studies to compare outcomes of amelanotic versus pigmented melanomas have not been performed. Treatment recommendations for AMM are identical to those for pigmented melanomas, although accurately defining clinical margins of the neoplasm often is challenging. Overall, a high index of suspicion and a low threshold to biopsy unusual clinical lesions ultimately may allow for earlier diagnosis and treatment of these frequently lethal malignancies. PMID- 9220552 TI - Desmoplastic neurotropic melanoma. AB - Desmoplastic neurotropic melanoma (DNM) is a rare variant of a spindle cell melanoma. The majority of these tumors occur on the head and neck of elderly patients. The rather variable clinical appearance (e.g. frequent lack of pigmentation) makes initial diagnosis often difficult. Histologically, DNM may show a lentiginous melanocytic proliferation with atypia and pleomorphic spindle cells in the dermis. Immunostaining for S-100 is usually positive although staining for HMB-45 is frequently absent. As with other melanomas, surgery is the first line treatment for DNM. Unlike other melanomas, however, survival for DNM may be better compared with other forms of melanoma. PMID- 9220553 TI - Variants of melanoma. AB - The current classification of malignant melanomas gives recognition to superficial spreading melanoma, lentigo maligna melanoma, acral lentiginous melanoma, and nodular types. In addition, neurotropic and desmoplastic types are recognized. The relativity inherent in the diagnosis of melanoma, provides the basis for the classification of melanomas on the basis of size. Lesions measuring 1 mm or less in vertical dimensions are unlikely to metastasize; they qualify as borderline melanocytic neoplasia of indeterminant malignant potential. The current classification has little relevancy to the category of variant nevi with the exceptions of malignant cellular blue nevus and melanoma arising in giant congenital nevi. A classification of variant melanomas as related to variant nevi is proposed. From a different perspective, a classification of melanomas with attention to nesting and cytological patterns in vertical growth is proposed: this alternate approach gives recognition to lesions that might otherwise be classified as "nevoid" melanomas. It also provides a default category for lesions that might otherwise be assigned to the Spitz nevus-like category. All of these tools for the manipulation of the real and virtual images of melanomas have been emphasized in the concept of minimal deviation melanoma. PMID- 9220554 TI - Surgical treatment of malignant melanoma. AB - The surgical treatment of malignant melanoma is dependent on early diagnosis and a thorough understanding of the options available to patients who present with more deeply invasive lesions. There has been a shift in therapy of primary skin lesions to using narrower (1-2 cm) margins of excision. Decisions regarding treatment of the regional lymph nodes are predicated upon understanding the overall risk of metastatic spread and delineating the pattern spread of the malignant cells. The concept of intraoperative lymphatic mapping can greatly aid the clinician in deciding (1) which lymph node group is the primary drainage basin for any particular area of skin, and (2) which lymph node is the first node to receive lymphatic drainage from any given area (the so-called "sentinel lymph node."). Surgical resection of metastases can result in prolongation of survival in carefully selected patients. PMID- 9220555 TI - New approaches to treating advanced melanoma: adjuvant treatment of high-risk primary melanoma and boron neutron capture therapy. AB - Malignant melanoma is associated with an excellent long-term prognosis when detected and treated at an early stage. Surgery alone is sufficient for patients with thin melanomas. Patients with thicker tumors, who are at higher risk for metastasis, may benefit from additional therapy beyond surgical removal of the tumor. Adjuvant therapies are designed to reduce the risk of melanoma recurrence. In particular, the immunotherapies, which boost the host's immune response to the cancer, are proving to be valuable adjuncts to surgery. Once melanoma has metastasized to a visceral organ, the prognosis is poor. Metastatic melanoma has a poor response to most conventional treatments. Boron neutron capture therapy is a novel approach to cancer management. It is a binary therapy combining low energy, nonionizing neutron irradiation with a stable isotope of naturally occurring boron avidly absorbed by the tumor. The combined approach currently under investigation results in highly selective tumor destruction. PMID- 9220556 TI - Skin self-examination for melanoma--another golden rule. AB - Melanoma is likely to become the most important cancer of the 21st century. Health education can improve the prognosis of this cancer. The goal of melanoma education is to reduce the rising incidence and mortality from melanoma. Skin self-examination is an important means of promoting awareness. Marketing the excellent health habit of inspecting our pigmented lesions on Melanoma Monday and several times annually requires teaching the public who is at increased risk, the signs of in situ and early melanoma, and the action to be taken if a suspicious spot is found. A skin scan for melanoma is a splendid health habit for every American. PMID- 9220557 TI - Substance use among homeless, immigrant, and refugee populations: an international perspective. Introduction. PMID- 9220558 TI - Substance use among street children in Honduras. AB - The hypothesis that drug use among Honduran street children is a function of developmental social isolation from cultural and structural influences is examined. Data from 1,244 children working and/or living on the streets of Tegucigalpa are described, separating "market" children from "street" children. The latter group is then divided into those who sniff glue and those who do not to identify salient distinguishing factors. An OLS regression of drug usage on these variables results in a model that explains 75% of the variance, where family relations, length of time on the street, and delinquency are the most important factors. PMID- 9220559 TI - Southeast Asians: Asian-Pacific Americans at risk for substance misuse. AB - Projections indicate that by the year 2000 over a million Cambodians, Laotians, and Vietnamese will be living in the United States. There is sparse information relative to the use of substances by these groups due to the absence of national prevalence data. The combined stressors that these refugee groups have faced puts them at high risk for substance misuse. Southeast Asians infrequently use substance misuse and mental health services, which has been perceived as a lack of need for services by these groups. In reality, there is a critical shortage of culturally-appropriate treatment and intervention programs as the prevalence of substance misuse increases in these populations. PMID- 9220560 TI - Ethnic differences in substance use and alcohol-use-related mortality among first generation migrants to England and Wales. AB - Epidemiological studies among migrant ethnic groups are potentially important as a way to provide insight into the relative importance of genetic, cultural, and socioeconomic factors in the etiology of substance use disorders. This paper summarizes prior United Kingdom studies of the prevalence of substance-use associated problems in different ethnic groups before analyzing trends in recent mortality data by country of birth. On this evidence, rates of alcohol-related mortality may be marginally higher for those born in the Caribbean than for the native British, but are substantially raised for those born in Ireland and the Indian subcontinent. There is some indication that rates for the Caribbean and possibly the Irish groups have risen more rapidly than for the national population over a 12-year period. These differences in mortality rates seem to have arisen for complex reasons. PMID- 9220561 TI - Substance use patterns among homeless migrants and nonmigrants in Chicago. AB - This paper uses data abstracted from 465 client records randomly selected from the current files of a downtown medical clinic in Chicago, Illinois to examine substance use among 85 immigrant and 380 nonimmigrant homeless and "at-risk" homeless adults. Immigrants to the United States reported lower levels of current cigarette, alcohol, and drug use compared to nonimmigrants in the sample. Immigrants were also less likely to report potential substance user treatment needs. Immigrants, however, did not differ from nonimmigrants in the reported quantity or frequency of cigarette and alcohol use. Drug use among homeless immigrants was also higher than estimates of misuse in the general population. The literally homeless in the sample reported higher levels of substance use compared to "at-risk" homeless. The literally homeless were also more likely to report higher levels of consumption and were more likely to have potential substance user treatment needs. The role of stress in the etiology of substance misuse among homeless immigrants and nonimmigrants is discussed. Implications for the treatment of these diverse populations are also addressed. PMID- 9220562 TI - Drug problems among immigrants and refugees in The Netherlands and the Dutch health care and treatment system. AB - Immigration from former colonies and the influx of refugees whose disadvantaged situation in society often leads to drug problems is described. The Dutch Government provides for a wide variety of services for drug users which include programs for minorities like the Surinamese, Moluccan, Moroccan, and Turkish communities. However, they are either executed poorly or are based too much on "White" methods. An account of a survey about the current situation of Black Surinamese women in the Netherlands is given, and the situation of refugees in the Netherlands as it relates to drug use is described. PMID- 9220563 TI - Homelessness and vulnerability among adults with and without alcohol problems. AB - While many works compare traits of homeless adults across levels of alcohol use, few specifically consider whether drinking status affects determinants of either homelessness or "vulnerability" to homelessness. This paper relies on a 1986 Chicago, Illinois sample (n = 535) to consider the potential contributions of resources, social network characteristics, disaffiliation, and mental health problems. Results suggest that resource problems may determine homelessness regardless of drinking status. But drinking-associated problems may raise the resource threshold for "vulnerability," reduce the protection afforded by social networks against both homelessness and "vulnerability," increase the deleterious impact of disaffiliation, and spur complicating mental health problems. PMID- 9220564 TI - "Substance abuse" disorders among runaway and homeless youth. AB - This study used systematic sampling methods to recruit a sample of 432 homeless youth from both service and natural "hang-out" sampling sites. According to DSM III criteria, the majority of respondents were classified as having an alcohol and/or illicit "drug abuse" disorder (71%). The results from multivariate logistic regression analyses indicate that cumulative length of time homeless is positively associated with risk for an "abuse" disorder. The implications of these findings and recommendations for service interventions are discussed. PMID- 9220565 TI - Drug use in Nepal: the view from the street. AB - This study uses qualitative research techniques to examine heroin use in Nepal. It explores the life histories of 16 heroin users in Kathmandu, the country's capital, emphasizing those who are street children or who are otherwise displaced. The cases document that the initiation of use in Nepal is a complex process which includes: certain personality traits; an early history of culturally acceptable use of alcohol, tobacco, marijuana, or hashish; peer influence; and the specific social setting of users. Outside (i.e., foreign) influences are included in the mix, but never as a single determinant of drug use. Factors specific to the Nepali scene include the traditional association of forms of marijuana with certain religious contexts and the availability of heroin. An additional factor is the poverty of the urban setting. The approach in Nepal to dealing with drugs primarily involves a realization of the role played by the interaction between personality and social setting in the fullest meaning of that term. PMID- 9220566 TI - Available resources for investigating substance use among homeless, immigrant, and refugee populations. PMID- 9220567 TI - Eye injuries in twentieth century warfare: a historical perspective. AB - With successive wars in the twentieth century, there has been a relative increase in injuries to the eye compared to injuries of other parts of the body. The main causes of eye injury have changed with advances in techniques and weaponry of warfare, with blast fragmentation injuries accounting for 50-80% of cases. Penetrating and perforating injuries are most common, and injuries associated with intraocular foreign bodies pose special diagnostic and management problems. Injuries are bilateral in 15-25% of cases. Injuries associated with chemical, nuclear, and laser weapons have distinct characteristics and epidemiology. Enucleation was commonly performed at the turn of the century, but incidence has declined with better understanding of the pathophysiology of ocular trauma, improved surgical techniques and sepsis control with antibiotics. Sympathetic ophthalmia appears to be uncommon and earlier fears of this complication seem to have been exaggerated. Timely evacuation to a surgical facility is important for a good visual prognosis and preservation of the globe. However, prevention of injuries with eye armor is ultimately the best management, and the need for a comprehensive eye protection program in the military cannot be overemphasized, especially since eye injuries pose important socioeconomic, as well as medical, problems. PMID- 9220568 TI - Ocular injury by mustard gas. AB - Sulfur mustard is a chemical warfare agent which was widely used during World War I and more recently in conflicts in the Middle East. This highly toxic compound causes severe dermal, gastrointestinal, respiratory and ocular injuries. It acts as an alkylating agent that induces structural changes and, hence, destruction of nucleic acids and proteins, impairing the cell's normal homeostasis and eventually causing its death. Sulfur mustard reacts rapidly with ocular tissues, and after a latent period of a few hours the patient starts suffering from severe eye pain, photophobia, excessive lacrimation and blindness. The injury, which is restricted to the anterior segment of the eye, may cause long-lasting incapacity in large numbers of casualties. Approximately 0.5% of the severely wounded victims may develop late complications which require prolonged ophthalmologic observation and therapy. In light of the ever-present threat of mustard chemical warfare against military and civilians, physicians worldwide should be aware of its grave effects and know how to care for its victims. PMID- 9220569 TI - Uveal lymphoid neoplasia: a clinical-pathologic correlation and review of the early form. AB - We report three cases of uveal lymphoid neoplasia that were diagnosed early in their course. One case exhibited a posterior form, presenting with progressive hyperopia from a serous-macular detachment and choroidal involvement along with retrobulbar involvement. This patient was treated with proton beam irradiation. Two cases displayed an anterior form, with fixed fleshy epibulbar masses resembling salmon patches, and choroidal involvement. The histologic findings from biopsy of these anterior masses are presented. One of these patients was treated with complete excision of the mass and double freeze-thaw cryotherapy of the scleral bed, and the other patient was treated with conventional photon beam irradiation. The clinical features of uveal lymphoid neoplasia in its early form are discussed. Evaluation and treatment strategies for these early forms of uveal lymphoid neoplasia are reviewed. PMID- 9220570 TI - Hypertensive retinopathy mimicking neuroretinitis in a twelve-year-old girl. AB - Bilateral disk swelling and marked peripapillary and macular exudates were found on routine ophthalmologic examination in a 12 1/2-year-old girl. Eleven months later, with persistent findings, her blood pressure was found to be extremely elevated. She had an Ask-Upmark kidney, a rare form of segmental renal hypoplasia. The Ask-Upmark kidney abnormality occurs primarily in young women and is associated with hypertension. The disk edema and retinopathy resolved after the hypertension was controlled. Hypertensive retinopathy can sometimes resemble neuroretinitis. PMID- 9220571 TI - Amaurosis fugax in the young. AB - A 36-year-old healthy woman developed amaurosis fugax of the right eye lasting 1 minute. Work-up revealed right carotid stenosis thought most compatible with an atherosclerotic plaque. A carotid endarterectomy was performed which corroborated the radiologic diagnosis. An embolic event from ipsilateral carotid artery disease should be considered as a cause of amaurosis fugax even in the young. PMID- 9220572 TI - Progressive visual loss and motility deficit. AB - A 63-year-old female with known stage III, low grade non-Hodgkin's lymphoma presented with progressive visual loss in the left eye and binocular diplopia in all positions of gaze. The left globe was almost immotile. Two MRI's of the orbit were interpreted as normal. Lumbar puncture did not reveal abnormal cytology. Although orbital apex involvement is uncommon in non-Hodgkin's lymphoma, the patient's clinical findings clearly indicated a lesion in this area, which was confirmed by a third MRI. Review of one of the initial films showed evidence of orbital apex involvement. To prevent diagnostic delay and unnecessary repeat imaging, the clinical diagnosis of orbital apex syndrome should be clearly communicated to the radiologist. Prompt recognition of orbital apex syndrome may improve visual outcome. PMID- 9220573 TI - Pulfrich revisited. AB - The Pulfrich phenomenon is a stereo-illusion resulting from latency disparities in the visual pathways. It is common after optic neuritis, but is also to be found with other conditions. The symptoms are often difficult for the patient to explain and for the physician to understand. Symptoms may be sufficiently disturbing to significantly interfere with a patient's life (e.g., prevention of driving). Treatment with the use of monocular tints is simple and effective. PMID- 9220574 TI - Prophylactic antibiotics in ophthalmic surgery. PMID- 9220575 TI - Transscleral argon-krypton laser coagulation. PMID- 9220576 TI - Generalized AA-amyloidosis in Siamese and Oriental cats. AB - During a 7 year period (1987-1994), 194 Siamese cats including a colour variant designated Oriental cat, were presented for post-mortem examination. Twelve of these animals (6.2%) were diagnosed with amyloidosis. Major gross pathological findings included enlarged pale livers with haemorrhages, pale and swollen spleens, and dilated intestines. Deposits of amyloid were found in these tissues. The amyloid was found to cross-react with anti dog AA-antiserum when examined with peroxidase antiperoxidase (PAP) staining (four cases). Amyloid fibrils were purified by the water extraction method and its major constituting protein (AA) was isolated by gel filtration. Amino acid sequence analysis of this protein from a Siamese cat and an Abyssinian cat revealed a significant difference between these breeds. In the Siamese protein AA two amino acid substitutions (46 R for Q and 52 V for A) were encountered. This finding indicates the existence of a new feline amyloid A protein occurring in the Siamese breed which differs from presently known (apoS)AA-proteins. Additionally, the pedigree analysis of affected cats suggests a familial trait. PMID- 9220577 TI - Cross-reactivity of monoclonal antibodies to defined human leucocyte differentiation antigens with bovine cells. AB - Thirty-seven subpanels of monoclonal antibodies (mAbs) included within the Vth International Workshop on Human Leucocyte Differentiation Antigens (Vth Workshop) were assayed for reactivity with bovine peripheral blood leucocytes. Sixty-five of the 772 mAbs (8.4%) stained bovine cells. mAbs from each of the 27 different CD groups that contained a mAb reacting with cattle were further investigated to compare the cellular expression of the antigen in cattle with that reported for the different CD antigens in humans. Two-colour immunofluorescence staining of the Vth Workshop mAbs against characterized bovine leucocyte subpopulation markers that identified monocytes, B cells, CD4, CD8 and WC1 +T cells were used for these analyses. Eighteen of the mAbs to different human CD antigens (CD11a, CD14, CD18, CD21, CD27, CD29, CD49a, CD49b, CD49d, CD49e, CD51, CD61, CD62L, CD62P, CD63, CDw78, CD98, CD100) stained bovine antigens with an almost identical cellular distribution to that reported in humans. This implies that these mAb react with the homologous cattle molecules. Nine mAbs (CD35, CD37, CD49c, CD50, CD54, CD66, CD81, CD88, CD102) stained bovine cells but the cellular distribution of the bovine antigen was different to that reported in humans implying either a different cellular distribution for these antigens in cattle or a reaction with a different molecule. The investigation has allowed the identification of several bovine homologues of human CD antigens that have not been previously defined in cattle and the cross-reacting mAbs will be valuable reagents for future investigations of bovine immunology. PMID- 9220578 TI - Longitudinal studies of immune function in cattle experimentally infected with bovine immunodeficiency-like virus and/or bovine leukemia virus. AB - The effects of single or dual infection with bovine immunodeficiency-like virus (BIV) and/or, bovine leukemia virus (BLV) on bovine immune function were examined over a 4 year period. Holstein calves were infected with BIV (four calves), BLV (five calves), BIV and BLV (five calves), or sham inoculated (three calves). Lymphocyte blastogenesis to mitogens, seven tests of neutrophil function, and mononuclear cell subset analysis by flow cytometry (BoCD4, BoCD8, BoCD2, BoWC1, sIgM+, and monocytes) were performed at regular intervals to 49 months post infection. These data were analyzed for main effects of each virus and interaction as a 2 x 2 factorial. BIV infected cattle had lower neutrophil antibody-dependent cell-mediated cytotoxicity and iodination responses during 2 of the 4 years post-infection (P < 0.05). BIV infection was not associated with any long-term significant changes in lymphocyte blastogenesis to mitogens or changes in mononuclear cell subset numbers in blood. There was a tendency for animals infected with BIV alone to have decreased lymphocyte blastogenic responses to mitogens, but this was not statistically significant. BLV infection caused an increase in total mononuclear cells with no dramatic shift in the relative proportions of the various subsets. Co-infection with BIV and BLV did not consistently cause a different response than either virus did individually. One BIV infected animal died of non-BLV lymphosarcoma 7 months after infection. All other animals had no unusual clinical signs. In summary, infection with BIV caused a significant, temporary decrease in neutrophil function with no consistent statistically significant alteration in lymphocyte blastogenesis or mononuclear cell numbers during the first 4 years after infection. BLV infection caused an increase in lymphocyte numbers, and there appeared to be no synergism between the viruses. PMID- 9220580 TI - Bovine CD8+ suppressor lymphocytes alter immune responsiveness during the postpartum period. AB - This study examined the immunoregulatory role of CD8+ lymphocytes during the postpartum period. Peripheral blood cells were isolated from postpartum and mid to late lactating animals. Flow cytometric analysis was performed to determine the frequencies of relevant cell populations. Depletion of CD8+ lymphocytes from whole cultures significantly decreased proliferation and cytotoxic ability of cells isolated from mid to late lactating animals. Enrichment of whole cultures with CD8+ lymphocytes further decreased their proliferative ability but pure CD8+ lymphocyte had increased cytotoxic activity. In contrast, neither depletion nor enrichment of whole cultures with CD8+ lymphocytes altered the already diminished proliferative responses of cells isolated from postpartum cows. No cytotoxic activity was observed by cells isolated from postpartum animals. Cultures from mid to late lactating cows mainly expressed IFN-gamma mRNA where as IL-4 mRNA was mainly expressed by cultures isolated from postpartum animals. Flow cytometric analysis revealed that CD8+ lymphocytes have a high level of activation and expression of the beta-chain during the postpartum compared with the mid to late lactating period. These data indicate that CD8+ lymphocytes are of the suppressor compared to the cytotoxic nature immediately following parturition. PMID- 9220579 TI - Bovine cytokine expression during different phases of bovine leukemia virus infection. AB - The potential role of aberrant cytokine production in the pathogenesis of bovine leukemia virus (BLV) was studied by analyzing cytokine mRNA expression in pokeweed-stimulated PBMLs of cows in different phases of disease progression. To analyze the mRNA, a semi-quantitative RT-PCR assay was developed. The RT-PCR assay was developed for detection of IL-2, -4, -6, -10, -12, IFN-gamma and actin using cDNA derived from phorbol-stimulated peripheral blood mononuclear leukocytes. Using a PCR specific for BLV tax, agar gel immunodiffusion and white blood cell counts, BLV-negative, BLV-positive aleukemic (AL), and BLV-positive persistently lymphocytotic (PL) cattle were identified. Peripheral blood lymphocytes cultured in vitro for 24 h in pokeweed mitogen were analyzed for cytokine production using the RT-PCR assay. Consistently elevated levels of IL-2 and IL-12 in AL and PL cattle in pokeweed mitogen-stimulated cells was detected, while IFN-gamma was elevated in the AL but not the PL cattle. PMID- 9220581 TI - Detection of Mycobacterium bovis infection in cattle using an immunoassay for bovine soluble interleukin-2 receptor-alpha (sIL-2R-alpha) produced by peripheral blood T-lymphocytes following incubation with tuberculin PPD. AB - After activation of T-lymphocytes with antigen there is an increase in the expression of interleukin-2 receptor-alpha (IL-2R-alpha) followed by the release of a soluble form of the molecule (sIL-2R-alpha) from the membrane of the stimulated cells. The present study investigates the novel use of the release of sIL-2R-alpha from activated T-lymphocytes as a marker of cell-mediated immunity (CMI) in cattle infected with Mycobacterium bovis. An enzyme immunoassay was used to detect sIL-2R-alpha produced following incubation of bovine peripheral blood mononuclear cells with mycobacterial antigens. Using this assay, 63/67 cattle naturally infected with M. bovis were identified whereas only 1/51 uninfected animals were considered to give a positive result. This assay is more convenient to use than lymphocyte proliferation assays which involve the use of radionucleosides. It should prove useful for monitoring the immunological activation of bovine T-lymphocytes in a variety of situations including the development of CMI responses in cattle to novel mycobacterial antigens or potential vaccines. PMID- 9220582 TI - Impaired specific immunoreactivity in cows with hepatic lipidosis. AB - In this study, hepatic lipidosis in cows was experimentally induced by offering an energy surplus during the dry period. Liver triacylglycerol (TAG) was 16% in the experimental group. In the control group fed the same diet in restricted quantities, liver TAG was about 7%. The animals of both groups were vaccinated with tetanus vaccine at Day 3 after parturition. It was demonstrated that the cows with high liver TAG percentages had lower humoral and cellular (P < 0.05) immunological responses compared with the animals with low liver TAG levels at Day 14 after vaccination. The results obtained in the high TAG group support the notion that the frequent occurrence of aspecific infections in cows with hepatic lipidosis may be due to impaired immunoreactivity. PMID- 9220583 TI - Effects of purified bovine whey factors on cellular immune functions in ruminants. AB - The immunomodulatory properties of bovine milk and whey have long been documented. The recent advance of whey protein fractionation technology has now allowed us to study the immunobiological properties of some highly purified components of whey, with a view to exploiting their possible industrial and biomedical applications. The effects of fractionated bovine whey proteins on cellular immune responses were therefore examined using a panel of in vitro assays. Both lactoferrin (LF) and lactoperoxidase (LP) were found to inhibit proliferation and interferon-gamma (IFN-gamma) production of ovine blood lymphocytes in response to mitogenic stimulation. However, their effects in a combined fraction or in whey protein concentrate (WPC) were either diminished or eliminated. LF and LP had no effect on lipopolysaccharide (LPS)-induced ovine blood lymphocyte proliferation, production of interleukin-1 beta (IL-1 beta) and tumour necrosis factor-alpha (TNF alpha) by ovine bronchoalveolar lavage (BAL) macrophages, major histocompatibility complex (MHC) Class II antigen expression by ovine BAL macrophages and bovine natural killer (NK) cell activity. However, alpha-lactalbumin (alpha LA) exhibited an enhancing effect on IL-1 beta production. It is noteworthy that as bovine whey fractions become progressively more purified, their modulatory effects on the immune response also become more clear-cut. The effects of LF, LP and alpha LA may be eliminated by their combination in whey or by other minor components of whey. Further investigation of industrial applications for whey proteins of high purity is warranted. PMID- 9220584 TI - Changes in the leukocyte phenotype profile of goats infected with the caprine arthritis encephalitis virus. AB - The proportions of different sub-populations of leukocytes in five healthy goats and five goats infected with the caprine arthritis encephalitis virus (CAEV) were examined using immunofluorescence and flow cytometry. A panel of monoclonal antibodies that identified a monocytegranulocyte marker (GMI); the CD4, CD8, IgM, MHC Class I, MHC Class II and T19 antigens, and the gamma delta (gamma delta) T cell receptor was used. We observed a significant (P = 0.016) reduction in the proportion of monocytes in the peripheral blood of infected (5.98%) compared with healthy control goats (9.92%). There was also a decrease in the proportion of CD4+ T lymphocytes that approached significance (P = 0.076) accompanied by a slight increase in the proportion of CD8+ T lymphocytes, in infected compared with uninfected animals. Consequently, three of the five infected animals had lower CD4:CD8 ratios than any of the healthy animals and two of these three ratios were inverted. Approximately 14% of T cells in the peripheral blood of healthy goats was identified as gamma delta T cells and all expressed the T19 antigen. A significantly elevated level of gamma delta T cells (P = 0.030) and an elevated level of T19 cells were observed in infected, compared with healthy animals. The proportion of leukocytes expressing surface IgM (B cells) was also elevated, although not significantly, in CAEV-infected compared to healthy controls. The changes in peripheral blood leukocyte subsets in infected goats suggest that immune responses to the infection are probably altered in these animals with eventual progression to severe disease and death. PMID- 9220585 TI - Expression of ovine interleukin-2 cDNA in Escherichia coli. AB - The expression plasmids pGEX-2T and pT7-7 were used to express ovine (Ov) IL-2 cDNA in Escherichia coli. The pGEX-2T vector contained glutathione-S-transferase (GST) as the affinity handle and resulted in high level expression of the GST-IL 2 fusion protein. However, only a small proportion of this fusion protein was present in the soluble fraction. The insoluble fraction was extracted with a detergent, sarkosyl, and even though a large amount of fusion product was obtained, it would not bind to glutathione beads efficiently. Thus, only low yields of biologically active rOvIL-2 were obtained. The yields were not significantly improved when other detergents were used for extraction except for a non-ionic detergent, Zwittergent 3-14, where there was a two- to three-fold increase compared with extraction with sarkosyl. An alternative vector, pT7-7 was used with a 6 x histidine tag followed by a thrombin cleavage site at the amino terminus of the mature ovine IL-2 protein to allow affinity purification by Ni NTA resin. A large proportion of the rOvIL-2 was partitioned to the insoluble fraction. This expression system was more useful than the pGEX-2T as large quantities of biologically active rOvIL-2 of at least 10 mg l-1 were obtained. The presence of the six histidine residues at the amino end of rOvIL-2 did not reduce its biological activity. Both systems yielded rOvIL-2 with a high specific activity of about 1 x 10(7) U mg-1 as measured by the ability to maintain proliferation of ovine ConA lymphoblasts. Recombinant OvIL-2 was active on bovine but not porcine ConA lymphoblasts. PMID- 9220586 TI - Porcine gastric mucosa associated lymphoid tissue (MALT): stimulation by colonization with the gastric bacterial pathogen, Helicobacter pylori. AB - The presence and features of mucosa associated lymhoid tissue (MALT), analogous to Peyer's patches, in the cardia of the lesser curvature of the porcine stomach are described. The gastric mucosa associated lymphoid tissue (gastric-MALT) is histologically distinct from gastric inflammation associated with colonization by normal gastric microflora and experimental bacterial colonization with a human gastric bacterial pathogen, Helicobacter pylori. The gastric-MALT consists of well-demarcated encapsulated and organized lymphoid tissue, intimately associated with overlying gastric epithelium, centered below the muscularis mucosae and drained by efferent lymphatics. Gastric-MALT was identified in all piglets studied including microbially sterile uninfected gnotobiotes; these structures were enlarged with age and local (gastric) antigenic stimulation. Significant (P < 0.05) expansion of the gastric-MALT occurred in H. pylori-infected gnotobiotic piglets. These distinct morphologic features and location in the cardia suggest that lymphoid elements in the gastric-MALT are involved in gastric antigen processing and regional lymphoid maturation, differentiation and proliferation in the stomach. PMID- 9220587 TI - Tissue chambers--a useful model for in vivo studies of cytokine production in the pig. AB - An in vivo tissue chamber model was developed to enable studies of local cytokine production and cellular events during inflammatory and immune reactions in the pig. Tissue chambers made of sialistic rubber tubing were surgically implanted in the subcutaneous tissue- and samples of tissue chamber fluid (TCF) and inflammatory cells were collected by aspiration with a syringe. To evaluate the model for local cytokine production, two cytokine inducers, polyribinosinic polyribocytidylic acid (poly I:C) and fixed Aujeszky's disease virus infected PK15 cells (ADV-PK15), were injected into the tissue chambers and samples of TCF were collected 0, 4, 8, 12, 24 and 48 h post injection. Poly I:C injections induced local production of interferon-alpha (IFN-alpha) as well as tumor necrosis factor (TNF) in the TCF but kinetic differences in the production of the cytokines were noted. Poly I:C also induced an increase in cell numbers in the TCF, mainly due to increased neutrophil numbers. Injections of ADV-PK15 induced local IFN-alpha production in the TCF as long as the pigs were serologically negative to ADV. Immunofluorescence and in situ hybridization techniques could be applied for characterization of TCF cells. Moreover, cells recovered from the tissue chambers were viable and could be used in functional in vitro tests. Taken together, this tissue chamber model could prove very useful in in vivo studies of inflammatory/immune responses and cytokine production in the pig. PMID- 9220588 TI - Monoclonal antibodies to a high molecular weight isoform of porcine CD45: biochemical and tissue distribution analyses. AB - This report describes the obtention and characterization of two monoclonal antibodies (mAbs), 6E3/7 [mAb 6E3/7 was submitted to the Second International Swine CD Workshop, where it has been assigned to CD45R] and 3C3/9, which recognize the isoform of highest molecular weight of porcine CD45. This conclusion is based on their cell reactivity and tissue distribution, identical to that reported for the human high molecular weight isoform of CD45, and on data from immunoprecipitation and immunoblotting analyses which show that these mAbs react with the largest polypeptide of those precipitated by mAb 2A5, that recognizes an epitope shared by all CD45 isoforms. These mAbs react with 60% of peripheral blood mononuclear cells (PBMC) but not with alveolar macrophages, granulocytes, platelets or erythrocytes. Antigen expression on PBMC is heterogeneous and is reduced after in vitro activation with mitogens. B cells and CD8+ T cells express more antigen than CD4+ T cells. Using immunoperoxidase techniques, the antigen was detected on B cell areas of lymph nodes and Peyer's patches, and on a subpopulation of medullary thymocytes. These mAbs will be useful reagents for functional and phenotypic analysis of porcine lymphoid cell populations by flow cytometry and immunohistochemistry. PMID- 9220589 TI - An homologous in vitro assay to detect lymphokines released by PHA-activated duck peripheral blood lymphocytes and spleen cells. AB - When stimulated with phytohaemagglutinin duck lymphocytes released lymphokines which were detected by their ability to maintain the proliferation of duck lymphoblasts using an in vitro assay similar to that previously developed with the mammalian system to measure IL-2. The inability of duck lymphokines to maintain the proliferation of mammalian lymphoblasts (mouse) indicated that there was no functional homology between duck and mammalian lymphokines. However, duck lymphokines did maintain the proliferation of chicken lymphoblasts indicating functional homology of these growth factors between these two species. The duck lymphokine maintenance assay is a simple and reliable test, and should be useful as an in vitro assay for the detection of factors released by antigen-specific lymphocytes when cultured in the presence of viral antigens. PMID- 9220590 TI - Development and efficacy of a vaccine against Streptococcus iniae infection in farmed rainbow trout. AB - Formalin killed bacteria were used as a vaccine against Streptococcus iniae infections in farmed rainbow trout. A single intraperitoneal injection of this vaccine in trout resulted in specific antibody production detectable for 6 months. Trout vaccinated at 50 g were protected under laboratory (experimental disease) and field conditions (natural disease) for at least 4 months against S. iniae infection. Passive transfer of S. iniae specific antibodies conferred protection. Under field conditions, mortality of non vaccinated trout exceeded 50%, whereas mortality of vaccinated trout did not reach 5%. In addition, vaccinated trout under field conditions gained 20% weight when compared with non vaccinated fish. PMID- 9220591 TI - Evaluation of commercially available assays of neopterin and beta 2-microglobulin for the assessment of disease progression in FIV-infected cats. AB - Serum or plasma samples from cats at different stages of feline immunodeficiency (FIV) infection and from uninfected cats were tested using immunoassays designed to detect human neopterin and beta 2-microglobulin (beta 2M). The results obtained from the anti-human neopterin assay did not correlate with infection status, time post-infection, fCD4 count or clinical picture. Feline samples gave negative results in the anti-human beta 2M assay. The assay kits used in this study are not suitable for the determination of the effect of FIV infection on immune activation markers in the cat. PMID- 9220592 TI - Functional aspects of Aujeszky's disease (pseudorabies) viral proteins with relation to invasion, virulence and immunogenicity. AB - In the present review, the interaction of Aujeszky's disease (pseudorabies) virus (ADV) with individual susceptible cells and the entire host is presented. Special emphasis is put on how viral envelope glycoproteins control invasion and virulence. Furthermore, the importance of envelope glycoproteins in the induction of a protective immunity is discussed. PMID- 9220593 TI - Recent developments in latency and recombination of Aujeszky's disease (pseudorabies) virus. AB - Latency is a characteristic and fascinating part of the biology of alphaherpesvirinae, including ADV. Tissue explanation, blot hybridization, in situ hybridization and more recently PCR are the experimental methods used to demonstrate that latent infections consistently occur in ganglionic neurons and, at a lower level, in tonsillar and possibly other cells. In vivo reactivation of ADV, resulting in shedding of virulent ADV, has been demonstrated experimentally following administration of high doses of corticosteriods. To determine the influence of vaccination with currently used gene deleted vaccines on field virus latency load, it is essential to use quantitative latency detection methods. We have developed chemiluminescence-based quantitative PCR assays specific for gG and gE, and are currently using these to determine field virus latency loads in tissues of pigs vaccinated with one of several gene deleted vaccines. Recombination between ADV strains has been demonstrated both in vitro and in vivo and has raised concerns about the generation of gene deleted virulent ADV strains. Recent studies in a mouse model have shown that high concentrations of both strains have to be present at the same anatomical site for recombination to take place. This led to the conclusion that ongoing ADV eradication programs, based upon the use of gene deleted vaccines and differential serological testing, are not likely to be threatened by recombination between virulent ADV and gene deleted vaccine strains. PMID- 9220594 TI - Effect of the concentration of maternal antibodies on the neural invasion of Aujeszky's disease virus in neonatal pigs. AB - The degree to which maternally derived antibodies may affect neural invasion of Aujeszky's disease virus (ADV) in neonatal pigs was examined. One-week-old pigs with different levels of maternal immunity were inoculated intranasally with 10(7.0) TCID50 of the Ka strain. The invasion of the virus was studied in both the trigeminal neural pathway (nasal mucosa, trigeminal ganglion = 1st level, pons/medulla = 2nd level and cerebellum/thalamus = 3rd level) and the olfactory neural pathway (olfactory mucosa = 1st level, olfactory bulb = 2nd level and lateral olfactory gyrus = 3rd level) by virus titration and immunohistochemistry (IHC). In control pigs without specific antibodies, virus invaded all neuronal levels in both neural pathways. In pigs with a low concentration of maternal antibodies (SN-titer = 2-3), virus infected all neuronal levels in both neural pathways but, compared to the controls, virus titers were significantly lower (approximately 2 log10) in the trigeminal pathway. In pigs with a high concentration of maternal antibodies (SN-titer = 272-384), virus reached the 2nd neuronal level of the olfactory pathway while no neural tissue had been infected in the trigeminal pathway. Virus titers in the affected neuronal levels of the latter pigs were significantly lower than in the controls. IHC revealed, in non immune pigs, a fibroblast-mediated spread of the virus in the nasal lamina propria, and a local spread of the virus from neurons to their satellite cells in the trigeminal ganglion. Such a spread of the virus was rarely seen in the nasal mucosa and in the trigeminal ganglion of passively immune pigs. These findings suggest that, in the presence of maternal immunity, defence mechanisms operate at these sites. In conclusion, we can state that a correlation exists between the level of maternal immunity and the protection against invasion of ADV in the nervous system of neonatal pigs. PMID- 9220595 TI - Detection of pseudorabies virus genomic sequences in apparently uninfected 'single reactor' pigs. AB - With a pseudorabies virus (PrV) gB ELISA, performed on 480,000 pigs on 8,900 Swedish farms, approximately 1,300 cases were observed with only one single animal reacting positively. These animals were termed 'single reactors' (SR). In order to find explanations for this peculiar phenomenon, the presence of PrV was investigated in organs of immunosuppressed and non-immunosuppressed SR animals. The virus was not detected by immunohistochemistry, virus isolation or co cultivation. An in situ DNA hybridization test detected PrV gC gene sequences in the olfactory bulb of one sow. A nested polymerase chain reaction (PCR) assay revealed gB, gE and gD gene sequences of PrV in the tissues of trigeminal ganglia, olfactory bulb, tonsils and brain. The nucleotide sequences of the amplicons revealed 98 to 100% homology with the corresponding sequences of PrV. The large latency transcript (LLT) was not detected in the organs of the SR pigs. Transmission of the SR phenomenon to animals in contact or to the next generation was not observed. Considering the present observations and the facts that (i) PrV vaccination is not applied in Sweden; (ii) the SR animals occur not only in the South, but also in Northern Scandinavia, which has no history of PrV infection and (iii) viral reactivation was not observed under natural conditions or after experimental immunosuppression, it is concluded that the SR phenomenon should hardly be considered as a typical PrV latency. The present findings show that certain herpesviral genomic sequences exist in apparently uninfected individuals. PMID- 9220596 TI - The role of biotechnologically engineered vaccines and diagnostics in pseudorabies (Aujeszky's disease) eradication strategies. AB - Modern-day biotechnology has an almost unlimited number of possibilities for reducing the impact of hereditary and infectious diseases. To date one of its most visible and rewarding applications for veterinary medicine has been in the genetic engineering of vaccines and diagnostics to assist in the eventual eradication of pseudorabies (PR, Aujeszky's disease). In the following review we summarize some of the most pertinent issues relative to PR eradication and point out the present and potential role of biotechnology in achieving our goal. PMID- 9220597 TI - Vaccination strategies for improving the efficacy of programs to eradicate Aujeszky's disease virus. PMID- 9220598 TI - Study of the delivery of the gD gene of pseudorabies virus to one-day-old piglets by adenovirus or plasmid DNA as ways to by-pass the inhibition of immune response by colostral antibodies. AB - In the present study, it was shown that piglets with maternal antibodies, which had been primed with a replication-defective adenovirus that expresses the pseudorabies virus (PRV) glycoprotein gD and boosted with the Bartha vaccine strain at 10 weeks of age are equally protected clinically upon a challenge as piglets without maternal antibodies vaccinated with the same approach or with the Bartha vaccine strain alone. Priming with a plasmid that expresses gD was less efficient. PMID- 9220599 TI - Evaluation of parenteral vaccination methods with glycoproteins against Aujeszky's disease in pigs. AB - A comparative evaluation of vaccination methods with glycoproteins for the induction of immune responses and protection of the pig against Aujeszky's disease virus (ADV) was performed. Different vaccination routes (intradermal (i.d.) versus intramuscular (i.m.)), inoculation sites (the neck versus the back) and number of inoculation points (2 versus 6) per site were compared. Body weight (BW) changes and viral excretion after challenge were compared with virus neutralizing titers, antigen-specific IgG and IgA responses in serum and virus specific lymphoproliferative responses in peripheral blood during the immunisation period. According to BW changes better protection was obtained with six-point than two-point i.d. injections. i.d. vaccination in the back at six points gave similar results as i.m. vaccination in the neck but appeared inferior in the reduction of virus excretion. Regarding the immunological parameters, the virus-specific IgA response in serum gave the best indication for protection. It can be concluded that according to BW changes, six-point i.d. immunisation in the back and i.m. immunisation in the neck provided the best protection and that six point i.d. injections resulted in a better vaccination than two-point i.d. injections. PMID- 9220601 TI - Evaluation of serological tests for the detection of pseudorabies gE antibodies during early infection. AB - Two enzyme-linked immunosorbent assays (ELISAs) and a particle concentration fluorescence immunoassay (PCFIA) were compared for their ability to detect antibodies against pseudorabies virus (Aujeszky's disease virus) glycoprotein E (gE) in the early stages of infection in pigs previously vaccinated with gE deleted pseudorabies vaccines. Seventy pigs were included in the study. Five groups of 6 pigs each were vaccinated with one of 5 different pseudorabies virus (PRV) gE-deleted vaccines, and subsequently infected intranasally with 10(5.6) TCID50 of the Iowa 4892 pneumotropic strain of PRV. This entire procedure was repeated using 10(4.6) TCID50 of the Rice strain of PRV. Five unvaccinated control pigs were also challenged with each virus strain. Three control pigs died before seroconverting, leaving 67 pigs for comparison. Blood samples were drawn from experimentally inoculated pigs on the day of vaccination, the day of challenge, and on 4-10, 14, and 21 days postchallenge (DPC). Serology test sensitivity estimates and comparisons among tests were made for each sampling day. Results of this study demonstrated differences among the tests in the time from inoculation to initial antibody detection, and the time to detect 50% and 75% of the infected pigs. The average time until first detection of pigs as seropositive for gE antibodies by PCFIA was 7.5 DPC. The blocking ELISA detected pigs as seropositive an average of 8.8 DPC, and the indirect ELISA first detected gE antibodies by 9.3 DPC. Fifty percent of the pigs were detected as seropositive by days 7, 8, and 9 for the PCFIA, blocking ELISA, and indirect ELISA, respectively. Similarly, 75% of the pigs were detected as seropositive by days 8, 9, and 10 for the PCFIA, blocking ELISA, and indirect ELISA, respectively. All pigs were detected as seropositive by 14 DPC for all 3 tests. PMID- 9220600 TI - Effect of maternally acquired Aujeszky's disease (pseudorabies) virus-specific antibody in pigs on establishment of latency and seroconversion to differential glycoproteins after low dose challenge. AB - This study investigated whether (1) passively immune pigs could become latently infected after challenge with low doses of wild type pseudorabies virus (PRV) and (2) if seroconversion to PRV could be consistently detected using two commercially available differential diagnostic ELISAs. Three litters of piglets with passively acquired PRV serum neutralizing (SN) antibody (geometric mean titers 47.03 to 95.10) were challenged at 6 to 12 days of age with 236 to 500 TCID50 of Shope strain virus; pigs were vaccinated at 11 weeks of age with a commercially available genetically engineered vaccine (TK- gE- gG- Iowa S62 strain PRV). Vaccination was intended to reduce the risk of reactivation of latent infection resulting in spread of virulent PRV infection to previously uninfected pigs during the experiment. Vaccination at this age also approximated common field practices in infected herds. After 15 weeks, all challenged pigs were seropositive on the PRV glycoprotein (g or gp) E differential ELISA but were seronegative on the gG differential ELISA. All three challenge groups had pigs that were latently infected as evidenced by the detection of PRV DNA by polymerase chain reaction (PCR) assay of their trigeminal ganglia (TG). There was a significant inverse relationship observed for age at challenge and the proportion of PCR positive pigs in the group 15 weeks postchallenge (p = 0.0004). This trend was independent of the passively acquired PRV SN antibody titers at challenge. In this study, passively acquired antibody did not provide protection against establishment of latent infection in piglets after exposure to low doses of virulent PRV. These latent infections were detected serologically by only one of two available differential diagnostic ELISA. PMID- 9220602 TI - Evaluation of tests for detection of antibodies to Aujeszky's disease (pseudorabies) virus glycoprotein E in the target population. AB - Receiver operating characteristic (ROC) curves assess the quality of tests over the entire range of test signals. We compared the ability of an ELISA to detect antibodies to Aujeszky's disease (pseudorabies) virus gE in colostrum (test A) and in a single droplet of whole blood (test B) with the results obtained in serum (gold standard) in the target population by constructing and analyzing such curves. The area under the ROC curve, which is a quantitative measure of test performance, proved to be significantly (p < 0.01) smaller in test A than in test B or the gold standard. No significant differences in the area under the ROC curve were observed between test B and the gold standard. PMID- 9220603 TI - Economic analysis of alternative AD control programmes. AB - The threat imposed by its virulence brings a presumption that Aujeszky's disease (AD) must be controlled because potential losses are high. Viewed as an economic problem, the decision on whether and how to control AD hinges on comparing the costs of doing so with the benefits (in terms of reduced production losses) to be gained. Four strategies are considered: (a) doing nothing, (b) suppressing and maintaining the disease at low prevalence levels by vaccination, (c) suppressing to low levels and then eradicating by culling remaining positive animals and (d) eradicating in one step by means of a test-and-slaughter policy. The net economic merits of each strategy are examined using data derived from specific vaccination studies established in Germany and the Netherlands. A computer model is developed to estimate disease costs under different technical, epidemiological and economic assumptions, allowing the economically optimal strategies to be explored. In general no single strategy can be recommended as the 'best' for dealing with AD, since it depends on a host of factors relating to pig density, prevalence levels, production system, trade relations, etc. As usual, economic realities complicate the quest for operational simplicity in disease control. However, for the regions of high pig density studied the most economic AD control strategy is to lower herd prevalence by intensive vaccination before completing eradication by test and-removal of remaining positive animals. PMID- 9220604 TI - Mechanisms of transmission of Aujeszky's disease virus originating from feral swine in the USA. AB - To understand the possible mechanisms of transmission of Aujeszky's disease virus (pseudorabies or PRV) from a feral pig reservoir, intranasal infections were initiated in domestic pigs and in pigs from a herd derived from captured feral pigs. Virus strains originating from feral pigs and from domestic pigs were compared. Similar shedding patterns were obtained in both feral-derived and domestic pigs, however, virus strains from feral pigs were markedly attenuated. Virus could be isolated after acute infection from nasal secretions, tonsils and occasionally from genital organs. In studies of transmission of PRV by cannibalism, either latently infected or acutely infected tissue was fed to both domestic and feral-derived pigs. In two similar experiments, latently infected tissue did not transmit virus, but tissues from acutely infected pigs did transmit infection. Cannibalism was observed typically in both types of pigs older than 6 weeks of age. It was concluded that transmission of PRV originating from feral pigs can occur by several mechanisms including the respiratory route and by cannibalism of pigs that die of acute infection. Transmission of PRV from feral swine may, however, result in sub-clinical infection. PMID- 9220605 TI - Genital infection and transmission of pseudorabies virus in feral swine in Florida, USA. AB - Seventeen feral swine (FS) naturally infected with pseudorabies virus (PRV) and treated with dexamethasone (4 mg/kg body wt) on five consecutive days shed virus primarily from the genital tract and less frequently from the upper respiratory tract. The FS isolates were identified as PRV by virus neutralization with specific polyclonal antiserum and by direct immunofluorescence. Restriction endonuclease analysis with BamHI showed that representative samples from a total of 62 isolates were identical to each other, but differed in at least 5 DNA bands from the PRV Shope reference strain profile. DNA purified from FS isolates propagated in Vero cells or DNA extracted directly from genital swabs were amplified in the polymerase chain reaction using primers specific for the gpII (gB) gene of PRV. This amplification yielded a product of the expected size (200 bp), which specifically hybridized to a digoxigenin-labelled 30-mer probe complementary to an area within the region defined by the primers. In a transmission experiment, PRV was recovered from the vagina at 1 and 6 weeks after uninfected feral gilts were mixed with infected feral boars. PRV was not isolated from the upper respiratory tract of either gilts or boars. At eight weeks, 4 of the 5 gilts had developed low titer neutralizing antibodies to PRV. Our results indicate that PRV in FS is transmitted through sexual contact. PMID- 9220606 TI - Isolation and characterisation of an Aujeszky's disease virus naturally infecting a wild boar (Sus scrofa). AB - Isolation of Aujeszky's disease virus (ADV) from an injured, female wild boar (Sus scrofa), shot dead by hunters, in an area adjacent to the Abruzzo National Park is reported. The brain was submitted for attempted virus isolation following episodes of mortality in several dogs and cats fed with meat from the wild boar. Virus was isolated on first passage from the brain of the wild boar. The restriction fragment length polymorphism profile of the isolate was assessed as a type I. The role of stress in reactivating latent ADV in wild boars, the possibility of transmitting infection to endangered species such as bears (Ursus arctos), wolves (Canis lupus), wild cats (Felis silvestris) and lynx (Lynx lynx), present in the Abruzzo National Park and the possible role of wild boars as reservoirs for ADV is discussed. PMID- 9220607 TI - No massive spread of pseudorabies virus in vaccinated sow herds. AB - In this study, we quantified transmission of PRV in 99 sow herds in which the sows were vaccinated three times a year with strain 783 O/W and found that the reproduction ratio R was 0.66, which is significantly below one. This implies that massive spread of PRV cannot occur in such herds. PMID- 9220608 TI - Analysis of the relationship between seroprevalence of Aujeszky's disease and pig density within the different areas of Brittany. AB - Data of the first general serological screening for Aujeszky's disease in Brittany are analysed. Brittany is divided in 4 'departements' and 1,263 'communes'. Relationships between seroprevalence and pig density are analysed, using a spatial moving average, at the 'commune' level. Pig density is related to seroprevalence of Aujeszky's disease, but this relationship is very different from one area to another. PMID- 9220609 TI - Aujeszky's disease and the European Community. AB - The situation as regards AD in the European Union in August 1995 is described. The territory has been divided into three zones comprising free regions, regions where eradication programmes are in operation and the remainder. The criteria used for defining these areas are described as are the movement rules applicable to movement into each zone. Considerable activity is now taking place to eradicate this disease and further progress is expected in the near future. PMID- 9220610 TI - Eradication and control programmes against Aujeszky's disease (pseudorabies) in France. AB - In the present paper, an overview is given about Aujesky's disease (AD) eradication programmes currently applied in the different departments of France, together with the obtained results. PMID- 9220611 TI - Aujeszky's disease (pseudorabies) virus eradication campaign in The Netherlands. AB - The Dutch Aujeszky's disease virus (ADV) eradication campaign is based on vaccination with glycoprotein E deleted vaccines. In the first stage of the programme, that was started in September 1993, the transmission of ADV must be reduced sharply. Subsequently, the remaining sources of virus need to be traced and eliminated. During the final stage, vaccination should be forbidden. This paper summarizes the observations made during a field study on the eradication of ADV by vaccination and reports the design and preliminary results of the first stage of the Dutch eradication campaign. PMID- 9220612 TI - Computer simulation to support policy making in the control of pseudorabies. AB - A further integration of international markets makes a coordinated policy against contagious animal infections increasingly important. In the future, stricter demands are to be expected concerning the control and eradication of such infections. To anticipate these demands, a computer simulation model is created in which scenarios can be evaluated with respect to epidemiological and economic effects of the infections and control strategies. In this paper, the simulation model is described for Pseudorabies in swine. In the model, the population of herds is subdivided into two main herd types: breeding and finishing. Each herd is in one of 24 states per herd type. The states are based on (1) the reproduction ratio R which is the number of secondary cases caused by one infectious herd, (2) the prevalence for each value of R and (3) the expected number of infectious animals in an infectious herd within each prevalence range and for each R. The different values of R are based on experiments and field data in which different vaccination strategies were used. The transition matrix with the probabilities of every transition from one state to another is calculated on a weekly base. With this matrix the distribution of herds over states from week to week is derived. To include a dynamic element in the transition probabilities, the number of newly infectious herds per week is a function of animal and other contacts, including aerial, material and personal contacts. Calculations show that the infection in the Dutch swine population will not disappear without vaccination. With a vaccination scheme in which sows are vaccinated 3 times per year and fattening pigs 1 time per cycle the infection will ultimately be eradicated, but 2 vaccinations per cycle for fattening pigs are needed to eradicate the infection within an acceptable timespan (i.e. 2 to 3 years). The latter strategy will become compulsory in the Netherlands from October 1st 1995. PMID- 9220613 TI - General overview of PRRSV: a perspective from the United States. AB - Four years after the report of its discovery, porcine reproductive and respiratory syndrome virus (PRRSV) continues to challenge swine producers, veterinary practitioners, and animal health researchers in the United States. The prevalence of infection is high--60% to 80% of herds is a reasonable estimate- but the clinical effects of infection vary widely among farms. In many herds, infection is unapparent and productivity seemingly unaffected. Some infected herds report occasional respiratory disease outbreaks in young pigs, or periodic outbreaks of reproductive disease, and a few herds experience severe, chronic disease problems, particularly in young pigs. In these herds, secondary infections with viral or bacterial pathogens, particularly Salmonella choleraesuis, Streptococcus suis, or Haemophilus parasuis typically occur concurrently with PRRSV infections. Understanding why some herds undergo devastating episodes of clinical disease and others show no apparent effects is central to solving the problem of clinical PRRS for swine producers. Understanding the ecology and epidemiology of PRRSV is the key to preventing and controlling PRRSV in the future. The objective of this article is to review recent developments in these areas. PMID- 9220614 TI - Molecular characterization of Lelystad virus. AB - Lelystad virus (LV), the prototype of porcine reproductive respiratory syndrome virus, is a small enveloped virus, containing a positive strand RNA genome of 15 kb. LV is tentatively classified in the family Arteriviridae, which consists of lactate dehydrogenase-elevating virus (LDV), equine arteritis virus (EAV) and simian hemorrhagic fever virus (SHFV). These viruses have a similar genome organization and replication strategy as coronaviruses, but the size of the genome is much smaller (12-15 kb) and they have different morphological and physicochemical properties. The genome of LV contains eight open reading frames (ORFs) that encode the replicase genes (ORFs 1a and 1b), envelope proteins (ORFs 2 to 6) and the nucleocapsid protein (ORF7). Genomic comparison of European and North American isolates has shown that the structural proteins encoded by ORFs 2 to 7 vary widely. The amino acid sequences of ORFs 2 to 7 of North American strains share only 55 to 79% identical amino acids with those of European strains. Using polyvalent porcine anti-LV serum, gene-specific anti-peptide sera and monoclonal antibodies, we have identified six structural proteins of LV and their corresponding genes. These are: the 15 kDa unglycosylated nucleocapsid protein (N) encoded by ORF7, an 18 kDa unglycosylated integral membrane protein M encoded by ORF6, a 25 kDa N-glycosylated protein encoded by ORF5, a 31-35 kDa N glycosylated protein encoded by ORF4, a 45-50 kDa N-glycosylated protein encoded by ORF3 and a 29-30 kDa N-glycosylated protein encoded by ORF2. A nomenclature for these structural proteins is proposed. PMID- 9220615 TI - Comparative morphogenesis of three PRRS virus strains. AB - The morphogenesis of a Dutch PRRS field strain virus (Lelystad virus) was studied and compared to that of a U.S. field strain VR2332 and its attenuated vaccine strain JJ1882. Porcine lung alveolar macrophages (PLAM) and CL2621 cells were infected with high doses of virus (MOI = 10). At 4, 6, 9, 12, 18, 24, and 48 h post infection (hpi) cells were fixed for electronmicroscopy or for detection of viral antigens by immunoperoxidase staining. From 6 hpi on, viral antigens were detected in the cytoplasm and from 9 hpi on completely assembled virus particles could be detected in infected cells. The three strains were similar in assembly of new virus particles, envelopment at the smooth endoplasmic reticulum, and egress from infected cells. However, distinct differences were seen in replication time of the three strains in various cell types. The Lelystad virus replicated very fast and efficiently in PLAM while VR2332 and JJ1882 replicated preferably in CL2621 cells. JJ1882 replicated faster in CL2621 cells than VR2332 did, probably because of increased adaptation to the cell-line. Although the U.S. and European strains differ at the level of the genome, morphogenesis is not visibly altered. There is however a distinct difference in preferred cell type between the European strain and the two U.S. strains. PMID- 9220616 TI - Variation in open reading frames 3, 4 and 7 among porcine reproductive and respiratory syndrome virus isolates in the UK. AB - Previous studies using monoclonal antibodies (mAbs) have revealed antigenic variation among UK isolates of porcine reproductive and respiratory syndrome viruses (PRRSV) and the use of in vitro translation products has shown that this variation lies in the protein encoded by open reading frame (ORF) 3. This protein has been shown to be present in purified virion preparations, suggesting that it is a structural protein. The original objective was to investigate the degree of variation of ORF3 among a number of UK isolates of different mAb reactivity and diverse chronology by sequencing and to correlate this with the mAb reactivity, in an attempt to define conserved and variable antigenic sites. A number of PRRSV isolates, from different outbreaks in the UK between 1991 and 1994, were propagated in pig alveolar macrophages and RNA extracted. The ORF3 and ORF7 regions of the individual viruses were amplified by the polymerase chain reaction (PCR) and their sequences were determined using internal sense and antisense primers. A number of differences among the sequences were noted within specific regions of the ORF3, with a hypervariable area detected at the carboxyterminal end, in the area of overlap with ORF4. With the most divergent isolate, 9.5% of the 84 translated amino acids encoded by the area of overlap were different from Lelystad isolate, translating the sequence in both reading frames. In view of consistent changes elsewhere in the ORF that suggest a common ancestry among the isolates studied, we conclude that this region may be subject to rapid change in comparison to other regions studied, and therefore may be an area subjected to immunoselective pressure. PMID- 9220617 TI - Pathogenesis and clinical aspects of a respiratory porcine reproductive and respiratory syndrome virus infection. AB - Some pathogenetic and clinical aspects of the respiratory tract infection with porcine reproductive and respiratory syndrome virus (PRRSV) are discussed. The acute and persistent stages of PRRSV infection are treated separately. Special attention is given to the author's work on experimental dual infections with PRRSV and other enzootic porcine respiratory viruses. It was concluded that: (1) Studies on the interactions of PRRSV and its target cell, the pulmonary alveolar macrophage, are very scarce. So far, the hypothesis of impaired macrophage function has not been proven. (2) The possibility that PRRSV causes disease in growing pigs in combination with other viral or bacterial infections has been demonstrated experimentally. Variation in disease resulting from such combined PRRSV infections will probably hamper future research into disease mechanisms. (3) There remains a need for more data on the clinicopathological significance of the persistent stage of PRRSV infections. PMID- 9220618 TI - Porcine reproductive and respiratory syndrome virus: a persistent infection. AB - Persistent infection with porcine reproductive and respiratory syndrome virus (PRRSV) was shown in experimentally infected pigs by isolation of virus from oropharyngeal samples for up to 157 days after challenge. Four 4 week old, conventional, PRRSV antibody-negative pigs were intranasally inoculated with PRRSV (ATCC VR-2402). Serum samples were collected every 2 to 3 days until day 42 post inoculation (PI), then approximately every 14 days until day 213 PI. Fecal samples were collected at the time of serum collection through day 35 PI. Oropharyngeal samples were collected at the time of serum collection from 56 to 213 days PI by scraping the oropharyngeal area with a sterile spoon, especially targeting the palatine tonsil. Turbinate, tonsil, lung, parotid salivary gland, spleen, lymph nodes and serum were collected postmortem on day 220 PI. Virus isolation (VI) on porcine alveolar macrophage cultures was attempted on all serum, fecal and oropharyngeal samples, as well as tissues collected postmortem. Postmortem tonsil tissues and selected fecal samples were also assayed for the presence of PRRSV RNA by the polymerase chain reaction (PCR). Serum antibody titers were determined by IFA, ELISA and SVN. Virus was isolated from all serum samples collected on days 2 to 11 PI and intermittently for up to 23 days in two pigs. No PRRSV was isolated from fecal samples, but 3 of 24 samples were PCR positive, suggesting the presence of inactivated virus. Oropharyngeal samples from each pig were VI positive 1 or more times between 56 and 157 days PI. Oropharyngeal samples from 3 of 4 pigs were VI positive on days 56, 70 and 84 PI. Virus was isolated from one pig on day 157 PI, 134 days after the last isolation of virus from serum from this animal. Virus was isolated from oropharyngeal samples for several weeks after the maximum serum antibody response, as measured by IFA, ELISA and SVN tests. All tissues collected postmortem were VI negative and postmortem tonsil samples were also negative by PCR. An important element in the transmission of PRRSV is the duration of virus shedding. The results of this study provided direct evidence of persistent PRRSV infection and explain field observations of long-term herd infection and transmission via purchase of clinically normal, but PRRSV infected, animals. Effective prevention and control strategies will need to be developed in the context of these results. PMID- 9220619 TI - Effect of porcine reproductive and respiratory syndrome virus on subsequent Pasteurella multocida challenge in pigs. AB - This trial was conducted to evaluate the effect of Porcine reproductive and respiratory syndrome virus (PRRSv) on a subsequent challenge with Pasteurella multocida in pigs. Sixteen, 3-4 week-old piglets, from a PRRSv and Aujeszky disease virus (ADV) free herd were used. Animals were equally and randomly allocated in four groups which were treated according the following schedule: Group I: negative controls; Group II: inoculation with only PRRSV; Group III: inoculation with PRRSV and P. multocida; Group IV: inoculation with ADV and P. multocida (positive controls). PRRSV and ADV were inoculated intranasally, at the doses of 10(4.6) and 10(4.5) TCID50/ml, respectively. Five days later, pigs from groups III and IV were inoculated intranasally, with two ml of a 10(9) CFU/ml suspension of equal parts of P. multocida, strains A52 and A24. No lesions were observed in piglets of group I. Microscopically, interstitial pneumonia was identified in all piglets of groups II and III and 3/4 piglets from group IV. Bronchopneumonia was detected in 3/4 of the piglets from group III and in all animals of group IV which, additionally, showed meningo-encephalitis and purulent rhinitis. Macroscopically, only piglets of groups III and IV had lung consolidation. However, much lower pneumonic scores (2.3%) were observed in group III, where 3 of 4 piglets were affected. On the other hand, all piglets of group IV showed some degree of pulmonary consolidation, with a mean score of 13.7%. Based on these results, it appears that the role of PRRSV as a initiator of secondary diseases is still undefined, but is probably mild. There was no clear interaction between PRRSv and Pasteurella multocida under the conditions and strains tested here. PMID- 9220620 TI - Porcine reproductive and respiratory syndrome virus (PRRSv) interaction with Haemophilus parasuis. AB - The interaction of bacteria and virus has been well demonstrated in the pathogenesis of respiratory disease in swine. The interaction between porcine respiratory and reproductive syndrome virus (PRRSv) and Haemophilus parasuis has not been studied. We initiated studies to evaluate a possible effect of the PRRSv on the pathogenesis of polyserositis caused by H. parasuis. A group of 30 three week old piglets were distributed in 4 groups. Group I (10 pigs) was inoculated with PRRSv and H. parasuis. Group II (10 pigs) was inoculated with H. parasuis alone. Group III (5 pigs) was inoculated with virus alone and group IV (5 pigs) was inoculated with culture media. Lesions consisted of a severe fibrinous polyserositis affecting 7 of 10 animals in group II and a mild fibrinous pleuritis in 1 of 10 animals of group I. Three of ten animals dually infected with the two agents died during the course of the study. These animals had pulmonary congestion and focal lung hemorrhages. No other animals died from other groups. Group III and IV had no macroscopic lesions. Microscopically group III had interstitial pneumonia. Immunomodulating virus effect may explain the differences in terms of lesions severity between groups I and II. Septic shock was suspected as cause of sudden death. PMID- 9220621 TI - Dual infections of PRRSV/influenza or PRRSV/Actinobacillus pleuropneumoniae in the respiratory tract. AB - To study the effect of a previous porcine respiratory and reproductive syndrome infection (PRRS) of the respiratory tract on influenza virus and Actinobacillus pleuropneumoniae (App) infections, 3-week-old specific-pathogen-free (spf) piglets were intranasally infected with PRRS virus. One week later, when the lung alveolar macrophages of PRRSV infected pigs were lowest in number, a second infection was applied by intranasal aerosol of influenza virus H3N2 or by endobronchial instillation of a mildly virulent App. The first experiment consisted of two groups (only influenza infection or dual PRRSV/influenza infection). A second experiment consisted of 4 groups (only influenza infection, only PRRSV infection, dual PRRSV/influenza infection and uninfected controls). At day 2, 4, 14 and 21 after influenza infection, two pigs were killed and sampled for virological and histopathological examination. Influenza H3N2 virus caused only a mild inflammation of the smaller bronchioli. Previous PRRSV infection did not influence clinical signs during influenza infection. Next, we studied in two experiments the effect of dual PRRSV/App infection during the acute stage at two days after App infection. In a third experiment, the influence of PRRSV on more chronic stages of App infection was studied at two weeks after the App infection. At the end of the experiments, the pigs were killed. Lungs were ranked according to size and kind of the lesions. Lesions were cut and measured, samples were taken for virological and histopathological examination. Statistical analysis of the ranked lung-lesions in the first experiment showed a distinct but small effect of previous PRRSV infection on the development of App-lesions. In PRRSV infected pigs. App produced a more severe disease. The second and third experiment however failed to show any influence of the previous PRRSV infection on the App infection. We conclude that previous PRRSV infection of the respiratory tract of spf pigs does not necessarily enhance the severity of secondary infections of the respiratory tract. PMID- 9220623 TI - Field isolates of porcine reproductive and respiratory syndrome virus (PRRSV) vary in their susceptibility to antibody dependent enhancement (ADE) of infection. AB - Seventeen porcine reproductive and respiratory syndrome virus (PRRSV) field isolates, including isolate ISU-P, were evaluated for their susceptibility to antibody dependent enhancement (ADE) of infection mediated by antibodies raised against PRRSV isolate ISU-P. Progeny virus yields of ISU-P and 4 of 16 field isolates in porcine alveolar macrophages (PAM) were reduced following treatment with a concentration of antibody that neutralized ISU-P (p < 0.01). In contrast, the yields of 12 of 17 field isolates were enhanced (p < 0.01). Treatment of all isolates with a 10-fold lower concentration of this antibody significantly (p < 0.01) increased virus yields of all isolates in PAM. However, the degree of enhancement varied among the isolates when compared to the enhancement of the yield of ISU-P. While no differences in enhancement were observed among ISU-P and 9 field isolates, yield enhancement of 6 and 1 isolates were less than and more than the yield enhancement of ISU-P, respectively (p < 0.05). The degree of enhancement mediated by a high concentration of antibody raised against ISU-P was inversely proportional to the ability of the antibody to neutralize the isolates (r = 0.92). In contrast, no direct correlation (r = 0.32) was observed between the degree of enhancement mediated by a low concentration of antibody and the ability of the antibody to neutralize the isolates. These data suggest that the variability in the susceptibility of PRRSV isolates to ADE arise from quantitative and/or qualitative differences in the antigenic determinants associated with virus neutralization and/or ADE. The antigenic diversity and the wide range in the susceptibility to ADE that exists among field isolates indicate that ADE should be taken into consideration in the development of effective immunization strategies for PRRS. PMID- 9220624 TI - Hematological and immunological parameters of 4 1/2-month old pigs infected with PRRS virus. AB - 4 1/2-month old SPF pigs were experimentally infected with PRRS virus. Blood samples were collected with regular intervals up to day 35 post inoculation (PI). Serum was used for PRRS virus isolation and antibody detection and stabilized blood for total leucocyte counts, differential counts and characterization of lymphocyte subpopulations by flow cytometry analysis using monoclonal antibodies specific for porcine CD2, CD4 and CD8. After an initial viremic period of 1-7 days duration for individual pigs, PRRS virus was intermittently detected in pigs up to day 18 PI. All pigs had developed antibodies against PRRS virus by day 14 PI. Total blood leucocyte counts and lymphocyte counts were significantly decreased for a few days shortly after infection, but had returned to pre infection levels on day 8-10 PI. A major change in the distribution of lymphocyte subpopulations was observed on day 3 PI, where the percentages of CD2+, CD4+ and CD8+ cells were significantly decreased. However, the percentages of these lymphocyte subsets quickly returned to approximately pre-infection values. The observed changes of the parameters examined do not indicate long-term systemic immunosuppression of the infected pigs. PMID- 9220622 TI - Immunity to PRRSV: double-edged sword. AB - The immune system is a double-edged sword for porcine reproductive and respiratory syndrome virus (PRRSV) infection. On one edge PRRSV has a predilection for immune cells and the disease manifestations can be linked directly to changes in the immune system. PRRSV appears to replicate extensively, if not exclusively, in cells of the immune lineage, notably macrophages; the direct replication of which may lead to immunosuppression, precipitate secondary infection and/or mediate disease. On the other edge, the virus stimulates immunity post-infection that protects an animal from re-infection. A vast array of structural and functionally distinct antibody specific to PRRSV are generated following infection or vaccination. Discrete populations of functional antibodies appear at different times and possibly reflect reactivity to different PRRSV polypeptides. Cell-mediated immune responses specific to PRRSV can be detected in various exposed pigs as well. Thus, the immune system appears to be intimately involved in both the disease process and protection from disease. It is unclear at this state of understanding what immune compartment provides protective immunity. It is humoral (i.e. antibodies), selective functionally distinct populations of antibodies specific for selected PRRSV polypeptides or is cellular immunity essential for protection, or both. This review will attempt to summarize the current state of knowledge of the complex interaction of the immune system and PRRSV. PMID- 9220625 TI - Diagnosis of PRRS. AB - This paper reviews various diagnostic methods for the detection of porcine reproductive and respiratory syndrome (PRRS) virus or antibodies to PRRS virus reported during the period from 1991 to 1995. In addition, experience from a European Community Concerted Action and especially Danish experiences concerning serological tests are presented. It is concluded that, in general, serological diagnosis with a high specificity and sensitivity is easy to perform on a herd level. However, no serological test has proven to be suitable for individual animal certification. PMID- 9220627 TI - Epidemiology of porcine reproductive and respiratory syndrome (PRRS): an overview. AB - PRRS disease was first recognised in the USA in 1987 and in Europe in 1990 and since then the disease has spread widely throughout many pig-producing countries. After a severe epidemic phase, the infection has become endemic. The prevalence of infection is generally high in infected countries. However, in areas with a low density of pigs, infection may spread slowly and if infected animal movements are not significant, farm-to-farm spread can be controlled and prevalence of infection maintained at a low level. The PRRS virus (PRRSV) was completely unknown before 1986, and the question of its origin remains unanswered. The exact epidemiologic relationship between American and European strains of PRRSV is difficult to establish because different isolates appear to belong to two distinct sub-populations which are only distantly antigenically related. In the environment, virus survival is optimal when temperature is cold and when ultra violet light exposure is low (little sunshine). These conditions are easily attained in winter and that may explain why virus spread increases during this period. Pigs of any age (including wild boars) are the only animals known to be naturally infected with PRRSV. Relatively close contact between pigs is the primary factor in virus transmission. Aerial transmission is a second mechanism of spread, particularly in winter and particularly over distances of less than 3 km. A third route of transmission is via semen. The role of fomites is not clearly documented, however since the virus is excreted in faeces and urine, slurry should be considered as a potential source of contamination. Within herds, the virus spreads rapidly with up to 85 to 95% of pigs in a herd becoming sero positive within two to three months. Thereafter, virus activity persists for extended periods (several month to years). Nevertheless, some authors have reported spontaneous elimination of PRRSV from infected farms. For the future, there remain questions concerning the possible evolution of the disease (in terms of its sanitary and economic impacts), and the possible influence of vaccines on the epidemiological features of PRRS. PMID- 9220626 TI - Indirect fluorescent IgM antibody response of pigs infected with porcine reproductive and respiratory syndrome syndrome virus. AB - IgG and IgM antibody responses were examined by an indirect fluorescent antibody method in pigs following inoculation with different porcine reproductive and respiratory syndrome virus (PRRSV) isolates or a vaccine virus. Viremia was also examined in the pigs. The IgG antibody was first detected between 9 and 14 days post inoculation (PI) and maintained high titers for at least 7 weeks PI. No change in IgG antibody titers was observed when the pigs were reinoculated with PRRSV 35 days PI. IgM antibody was detected between 5 and 28 days PI in the pigs. Reinoculation at 35 days PI caused a short term rise of IgM antibody. Virus was isolated from sera collected between 2 and 21 days PI. The IgM antibody was detected regularly in sera collected during viremia and up to 1-2 weeks after the viremic periods. These results suggest that pigs with detectable IgM antibody are probably pigs with recent infection and that routine testing of IgM antibody in purchased breeding pigs from seropositive farms may be useful in identification of pigs with recent infection. PMID- 9220628 TI - Clinical signs and economic losses caused by porcine reproductive and respiratory syndrome virus in a large breeding farm. AB - In July of 1994 an acute onset of maternal reproductive failure occurred in a 2,330 sow farrow-to-finish farm. Clinical signs observed in the affected sows were typical for porcine reproductive and respiratory syndrome (PRRS). During the first 6 weeks of the epizootic 1,117 sows farrowed; 216 (19.33%) farrowed before the 110th day of gestation. The majority of piglets born before term died within a few days of birth and the mortality rate for term piglets increased to a maximum of 75.56% during the 5th week of the epizootic when 1,562 out of 2,067 piglets were either born dead or died prior to weaning. Preweaning mortality rates gradually returned to normal values within 16 weeks. The incidence of respiratory disease in the weaned and fattening pigs increased during this time. Although specific prophylactics against respiratory diseases were administered, the death rate doubled for the weaned and fattening pigs. PMID- 9220629 TI - A case-control questionnaire survey of risk factors for porcine reproductive and respiratory syndrome (PRRS) seropositivity in Danish swine herds. AB - Sixty-eight case herds seropositive to porcine reproductive and respiratory syndrome (PRRS) were compared to 128 seronegative controls in a double-blinded questionnaire survey. The study indicated no increased risk of PRRS seropositivity for herds using artificial insemination with semen from PRRS seropositive AI-stations. Also the herd-size was non-related to the risk of PRRS seropositivity, indicating that air-borne spread of PRRS may not have been a predominant feature in Denmark. Introduction of replacement breeding animals from seropostive breeding- and multiplying herds significantly increased the risk of a herd being PRRS seropositive, as did introduction of 25 kg pigs for feeding. PRRS seropositivity was in the farmers' opinions associated with abortions in sows, early farrowing, high postweaning mortality and low weight gain in fattening pigs. However, the reported frequencies of probelms were relatively low. PMID- 9220630 TI - Studies of porcine reproductive and respiratory syndrome (PRRS) virus infection in avian species. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) is a recently recognized virus of swine. As a newly emerging virus, much of the basic information regarding PRRSV is in the process of discovery. We report three experiments with PRRSV in birds, and a fourth experiment to evaluate the infectivity and transmissibility of avian-derived PRRSV in swine. Experiment 1 compared the susceptibility of Muscovy ducks, Mallard ducks, guinea fowl, and chickens to PRRSV. Birds were exposed to PRRSV (ATCC VR-2402) in drinking water and virus isolation was attempted from feces collected from cages. Based on the duration of fecal shedding of the virus, this experiment showed that Mallard ducks were particularly susceptible to PRRSV. Experiment 2 was done in mallards to corroborate and augment the observations of experiment 1. Virus was isolated from pooled mallard feces up to 25 days post exposure (PE) and from the intestinal contents of 8 of 20 birds euthanized on day 38 PE. No gross or microscopic lesions were observed in ducks collected between 0 and 15 days PE. Experiment 3 evaluated the infectivity and transmissibility of mallard-derived PRRSV in mallards. A cage of mallards orally exposed to PRRSV shed the virus in feces. Exposure of a second cage of mallards to feces from the first cage resulted in fecal shedding of PRRSV by birds in cage two. In turn, exposure to feces from the second cage led to fecal shedding by mallards in a third cage. Experiment 4 assessed the infectivity and transmissibility of mallard-derived virus in swine. Pigs intranasally exposed to PRRSV isolaed from mallard feces in experiment 2 became viremic, seroconverted by ELISA, and transmitted the virus to sentinel swine. Collectively, these studies show that the possibility exists for avian species to be involved in the epidemiology of PRRSV. This is the first report of PRRSV infection in a species other than swine. PMID- 9220631 TI - Monitoring of porcine reproductive and respiratory syndrome virus infection in boars. AB - A major concern exists on transmission of porcine reproductive and respiratory syndrome virus (PRRSV) via semen and effect of vaccination on PRRSV shedding in semen. Recent reports suggest that the virus can be transmitted by semen from boars infected experimentally or from natural sources. Seminal shedding, viremia, and changes in semen quality in boars with or without vaccination were examined. Nine boars were divided into three groups (three boars/group). Group I boars were vaccinated with 2 ml of RespPRRS vaccine (NOBL Laboratory) intramusculary and groups II and III were non-vaccinated. At 28 post-vaccination study days, group I and group II boars were challenged with virulent PRRSV VR-2332 at 2 ml of 10(4.0) TCID50 per boar intranasally. Group III served as non-vaccinated and non challenged control. Semen and serum samples were collected from -9 pre vaccination study days to 85 post-challenge study days and tested for the presence of PRRSV by virus isolation and reverse transcription-nested polymerase chain reaction (RT-nPCR). Prior to detection of PRRSV RNA from samples, conditions for RT-nPCR were optimized. Two primer sets, an external and an internal, were selected for RT-nPCR. The first round of PCR using an external primer set could detect 10 TCID50 of PRRSV/reaction. However, nested PCR could detect as little as 0.01 TCID50 of PRRSV/reaction. PRRS vaccine virus was not isolated from vaccinated pigs, but the vaccine virus RNA was detected from three boars, at day 6 to 15, 9 to 12, and 15 to 21 post-vaccination by RT-nPCR. Following challenge, two of non-vaccinated/challenged boars shed virus into semen up to 50 and 57 days post-challenge, respectively. The group I vaccinated boars did not shed virus into semen after challenge. The non-vaccinated/challenged group featured sperm abnormalities in the form of significantly increased incidence of proximal droplets and abnormal tails at 36-50 days post-challenge. The latter defect was observed to increase similarly in vaccinated/challenged boars as well. PMID- 9220632 TI - Strategies to control PRRS: a summary of field and research experiences. AB - Various methods for the control of PRRS virus have been published. The technology of nursery depopulation (ND) appears to effectively control the spread of virus between members of endemically infected populations. ND consists of a strategic adjustment in pigflow based on the presence of specific serologic patterns as detected by the indirect fluorescent antibody test. This pattern indicates a low seroprevalence of antibodies detected in the breeding herd and recently weaned piglets (< or = 10%), in contrast to a high (> 50%) seroprevalence in 8 to 10 week old piglets. ND has been carried out on swine farms in the US and results indicate improvements in nursery piglet growth rate and mortality levels. Three examples are provided in the following text. Recently a modified live virus vaccine (RespPRRS, NOBL Laboratories/Boerhinger Ingleheim) has become commercially available. It is currently approved for use in piglets from 3 to 18 weeks of age; however, potential for the use in adult animals is currently under investigation. PMID- 9220633 TI - Results of a control programme for the porcine reproductive and respiratory syndrome in the French 'Pays de la Loire' region. AB - The porcine reproductive and respiratory syndrome (PRRS) virus first entered the Pays de la Loire region in November 1992, with variable effects ranging from sub clinical seroconversion to severe reproductive failure and piglet mortality, and significant reduction of daily weight gains in finishing pigs. An epidemiological survey was carried out in February 1993. Since the infection prevalence was low (11 infected out of 2310 herds), the pig population was of medium density and the eradication programme of Aujeszky's disease had been successful in the Pays de la Loire region, it was decided (in March 1993) to undertake a control programme for PRRS. In 1993, introduction of infected pigs was known to be the most frequent source by which PRRS virus entered a herd. In the absence of vaccination, this source of virus introduction was reduced by a control programme applied to all members of the regional pig industry, through the impetus of the leaders of the Regional Sanitary Defence Confederation (FRGDS). The control programme was applied on purchased animals (sows, boars, piglets), artificial insemination centres and other environmental factors (people, vehicles, materials, slurry,...). Moreover, pigs from many infected herds were slaughtered. Results showed that in a context of low prevalence and limited spreading to nearby herds, efficient control of animal movements limited the infection spread. At the end of 1993, the PRRS prevalence was 2.7% in the region. Two years after the first outbreak, the PRRS infection could be considered as controlled since 98% of the herds remained free. In order to maintain this low infected status, the control programme was renewed. From this study epidemiological investigations have raised two major initial sources of infection, the use of contaminated semen and the introduction of infected pigs. Around an infected herd, serological screening is still running to detect infection in nearby herds. PMID- 9220634 TI - Efficacy of an inactivated vaccine for prevention of reproductive failure induced by porcine reproductive and respiratory syndrome virus. AB - This report describes the results of experiments with an inactivated oily vaccine containing per dose about 10(5.5) median tissue culture infectious dose (TCID50) of the Spanish strain of porcine reproductive and respiratory syndrome (PRRS) virus grown in porcine alveolar macrophages (PAMs). In order to evaluate the efficacy of the vaccine, two experimental infection routes were tested in sows; subsequent intranasal (i.n.) and intravenous (i.v.) (out of a total of 93 piglets born to 7 sows, 16% were mummified, 18.2% were weak and died within 48 h of birth, 37% were stillborn, 5.3% died between 2 and 7 days of age, 22.5% lived for more than one week) and intranasal alone (out of a total of 65 piglets born to 5 sows, 0% were mummified, 22.5% were weak and died within 48 h of birth, 40% were stillborn, 4.6% lived for more than one week). I.N. alone was selected to evaluate the efficacy of the vaccine because this is the natural route of infection. A number of experiments were conducted to test the immunogenicity of the vaccine. In general, after challenge with the homologous strain, protection in vaccinated sows was high (at least 70% of the piglets were born alive and healthy), whereas protection in unvaccinated sows was low (only 10% of the piglets were born alive and healthy). Vaccinated animals devoid of antibodies by immunoperoxidase monolayer assay (IPMA) at the time of challenge were still protected at experimental infection. PMID- 9220655 TI - Allogenic transplantation of mobilized peripheral blood progenitor cells: towards tailored cell therapy. AB - Mobilized peripheral blood prognitor cells (PBPC) are increasingly being used instead of bone marrow for allogeneic transplantation. The present article will briefly review important aspects of allogeneic PBPCT including, donor safety, timing of leukapheresis, factors influencing the yield, cellular composition of PBPC allografts, hematopoietic capacity of allogeneic PBPC, and graft-versus-host and graft-versus-leukemia activities. It will particularly focus on the perspectives opened by PBPC for graft engineering and cell therapy. PMID- 9220656 TI - Rare but important adverse effects of all-trans retinoic acid in acute promyelocytic leukemia and their management. AB - Several adverse effects have been reported to occur after clinical application of all-trans retinoic acid (RA) in acute promyelocytic leukemia (APL). Except for severe side effects including retinoic acid syndrome, the mechanism of action of RA on adverse effects remains unclear. Here we describe some rare adverse effects and their management. We reviewed the English literature, and we added our cases of endocrine and metabolic adverse effects, such as hypercalcemia, male infertility, bone marrow necrosis, fibrosis and acute pancreatitis. We also described our cases of thromboembolic events, RA-dependent growth of pathologic cells including Sweet's syndrome, erythema nodosum, hyperhistaminemia, granulomatous proliferation, and mild cases of pulmonary complications. In addition, we reviewed the efficacy of RA administration for other types of leukemia or myelodysplastic syndrome. RA and chemotherapeutic agents might induce complete remission, but we obtained a response in only one case of M2 in the third relapse. During RA administration the patient should be monitored for these adverse effects, and early diagnosis and appropriate treatment are important. PMID- 9220657 TI - The revised classification of von Willebrand disease including the previously masqueraded female hemophilia A (type 2N) AB - The revised classification of von Willebrand disease was proposed by the working committee (Chairperson: J.E. Sadler) and approved by the Scientific and Standardization Committee/the International Society on Thrombosis and Hemostasis in 1993. It consists of three major types; type 1 with decreased level of normal von Willebrand factor (VWF) and type 3 with complete deficiency of VWF as quantitative abnormality, and type 2 with qualitative anomaly of VWF subcategorized into 2A, 2B, 2M and 2N, respectively, depending on the functional abnormalities (decrease of GPIb binding with defect of VWF large multimers, increase of GPIb binding, decrease of GPIb binding with VWF large multimers, decrease of FVIII binding). This classification is frequently quoted in the USA and Europe. We introduce this classification and review again the structural abnormalities of VWF, including the previously masqueraded female hemophilia A (type 2N). PMID- 9220658 TI - The occurrence of beta-thalassemia mutation and its interaction with hemoglobin E in the eastern India. AB - The distribution of variant hemoglobin in India has been related to various ethnic groups among other factors. beta-Thalassemia is the most frequent monogenic disorder in the country. Analysis of hemoglobin of 435 cases from Eastern India was performed by electrophoresis and by other quantitative methods. Analysis of the beta-globin gene of 112 cases used ARMS (amplification refractory mutation system)-PCR (polymerase chain reaction) techniques showing that IVS-1 nt 5 (G-->C) is the most prevalent mutation in populations from Eastern India. IVS-1 nt 5 (G-->C) interacts with the codon 26 (G-->A) mutation to produce E beta thalassemia phenotype in the samples from West Bengal, India. PMID- 9220659 TI - GM-CSF- and IL-3-dependent CD34 expressing myeloid cell line (SAS-1) established from CD7 and CD34 expressing acute myeloblastic leukemic cells. AB - A novel factor-dependent human myeloid leukemia cell line (SAS-1) was established from a 69-year-old Japanese male suffering from CD7 and CD34 expressing acute myeloblastic leukemia (AML M2 in FAB classification). Morphological and cytochemical staining showed that SAS-1 cells were round with basophilic cytoplasm which is positive for peroxidase. Analysis of surface markers revealed that SAS-1 cells were myeloblasts derived from an immature progenitor origin, which express CD34. The consensus karyotype of the cell line was 41 XY 5q-, -7, 11p-, 12p+, -13, -14, -16, -17, -19, -22, with two markers. The proliferation of SAS-1 cells was dependent on the presence of either granulocyte-macrophage colony stimulating factor (GM-CSF) or interleukin-3 (IL-3), and GM-CSF- and IL-3-induced proliferation was dose dependent. Neither, stem cell factor (SCF) nor granulocyte colony-stimulating factor (G-CSF) alone supported the growth of SAS-1 cells, but supported their viability for more than 4 days and arrested them in the G0/G1 phase. SCF also enhanced GM-CSF- or IL-3-induced growth, but other cytokines did not have this synergistic effect. Clonogenic assays revealed that SAS-1 cells formed 36.0 +/- 5.7 or 41.5 +/- 0.7 colonies/1000 cells in the presence of GM-CSF or IL-3, respectively. SCF also increased the number of colonies formed by GM-CSF or IL-3 treatment, while SAS-1 cells did not form colonies in the presence of SCF alone. SAS-1 cells may prove to be a useful tool for studying the regulation of the cell cycle, myeloid proliferation, and differentiation. PMID- 9220660 TI - Neutropenic enterocolitis in adult leukemias. AB - Neutropenic enterocolitis is a frequent autopsy finding in adult patients with acute leukemias. The predisposing factors other than neutropenia are not clear. There are also contradictions about treatment. Therefore, this entity still presents a diagnostic and therapeutic dilemma for clinicians. This retrospective study was performed to determine the incidence of neutropenic enterocolitis in adult leukemic patients, to examine the possible risk factors, clinical characteristics and treatment outcome. The pathogenesis is also discussed considering clinical and laboratory findings of the patients. The incidence of neutropenic enterocolitis was 6.5% for acute myeloid leukemia and 4.6% for acute lymphoblastic leukemia. The mean absolute neutrophil count at diagnosis was 48/mm3 (median: 0/mm3). The median duration of severe neutropenia (absolute neutrophil count < or = 500/mm3) on follow-up before the diagnosis was 32 days. Thirteen out of 20 patients had received antineoplastic drugs within the last 12 days, but 2 had not. Fourteen patients had signs of peritoneal irritation and 3 of them underwent surgery. The others received supportive measures, i.e. bowel rest, intravenous fluids, combinations of wide spectrum antibiotics, transfusions, hemodynamic supports and nasogastric decompression, if necessary. All 3 patients who underwent surgery survived, whereas only 1 of the 11 who received other treatments did. Six patients without signs of peritonitis were treated with antibiotics and the mentioned supportive measures. Four survived, but the others died due to sepsis. In conclusion, considering some recent reports that stated good outcome with conservative measures in the presence or absence of peritonitis, there is still debate regarding the optimal choice of treatment. The importance of early diagnosis cannot be overemphasized. Signs of peritoneal irritation indicate a life threatening condition. Surgery can be performed successfully in such patients. Long duration of neutropenia may be an important risk for neutropenic enterocolitis. PMID- 9220661 TI - Pharmacokinetic studies of intravenous glycosylated recombinant human granulocyte colony-stimulating factor in various hematological disorders: inverse correlation between the half-life and bone marrow myeloid cell pool. AB - The pharmacokinetics of an intravenous bolus dose of glycosylated recombinant human G-CSF (rhG-CSF) was examined in 15 patients with various hematological disorders and 3 normal volunteers. The elimination half-life of rhG-CSF varied with the disorder. The half-life of an initial dose of rhG-CSF (2 micrograms/weight kg) was significantly prolonged in patients with aplastic anemia (2.7 +/- 0.3 h, n = 3) and myelodysplastic syndrome-refractory anemia (2.0 +/- 0.3 h, n = 3) when compared with those in normal controls (0.9 +/- 0.5 h, n = 3). In contrast, in patients with acute myelogenous leukemia which was overt leukemia from myelodysplastic syndrome-refractory anemia with excess of blasts in transformation, the half-life was shortened after chemotherapy (0.2 +/- 0.1 h, n = 3). The half-life of rhG-CSF in 2 patients with acute lymphoblastic leukemia in complete remission was prolonged (2.0 and 2.7 h) at the time of marrow suppression after chemotherapy and then shortened (0.5, 1.0 h, respectively) in the recovery phase. The half-life of rhG-CSF was very weakly, inversely correlated with absolute neutrophil count in blood (n = 24, r2 = 0.32, P < 0.01), and was inversely correlated with the absolute count of bone-marrow myeloid cells (nucleated cell count in bone-marrow aspirates x the percentage of myeloid cells/100) of patients with aplastic anemia and myelodysplastic syndrome refractory anemia (n = 12, r2 = 0.63, P = 0.002). These results suggest that the half-life of intravenously administered rhG-CSF (2 micrograms/kg) reflects the size of the myeloid cell compartment in vivo, and support the hypothesis that receptor-mediated consumption mainly accounts for the clearance of exogenous G CSF. PMID- 9220662 TI - Demonstration of functionally distinct human polymorphonuclear leukocyte fractions by simultaneous measurement of phagocytosis and oxygen radical generation. AB - Phagocytosis and oxygen radical generation by human polymorphonuclear leukocytes (PMNLs) were studied by a two-color flow cytometric analysis, where the red fluorescent product(s) of hydroethidine was used as an indicator of intracellular generation of oxygen radicals and opsonized zymosan (OZ) as an indicator of phagocytosis. Unstimulated cells formed a single population of cells without any significant fluorescence. PMNLs stimulated by OZ exhibited a high red fluorescence. Most PMNLs phagocytosed OZ and generated oxygen radicals when stimulated by zymosan particles opsonized with human AB serum at concentrations of more than 10%, whereas three distinct subpopulations (designated as R1, R2 and R3) appeared when stimulated by zymosan particles opsonized with 3% serum; R1 cells enhanced neither green nor red fluorescence, R2 cells enhanced green fluorescence but not red fluorescence, and R3 cells enhanced both green and red fluorescence. The R2 cells completely disappeared by the addition of Trypan blue. Most of the R3 cells disappeared by the addition of cytochalasin B. These findings on the three fractions were confirmed by the observation under fluorescence microscopy of cells in each fraction obtained by sorting. In conclusion, PMNLs could be separated into three functionally distinct subpopulations when stimulated by zymosan particles opsonized with a suboptimal concentration of serum. PMID- 9220663 TI - Differential induction of apoptosis on human lymphoblastic leukemia Nalm-6 and Molt-4 cells by various antitumor drugs. AB - To investigate how chemotherapy agents interact with the leukemic cell death pathway, we examined apoptosis of human lymphoblastic leukemia cells (Nalm-6 and Molt-4) treated with various anticancer drugs (etoposide (VP-16), camptothecin (CPT), adriamycin (ADR), cytosine arabinoside (Ara-C), methotrexate (MTX), 6 mercaptopurine (6MP), cyclophosphamide (CPM), vincristine (VCR) and prednisolone (PRD)) by flow cytometric procedures. The proportion of apoptotic cells was estimated from the presence of cells with a fractional DNA content in the DNA histograms after the incubation of drug-treated cells with a DNA extraction buffer. Treatment with Ara-C, CPT, VP-16 and ADR resulted in rapid apoptosis with 40-60% apoptotic cells by 8 h. Treatment with MTX, VCR, 6MP and PRD induced no apparent apoptosis until 12 h, but further treatments with these drugs resulted in apoptosis with 50% (MTX), 20-30% (6MP and VCR) and 5-10% (PRD) apoptotic cells, respectively, at 24 h. CPM induced apoptosis with 10-20% apoptotic cells at 10(-6) M, but higher doses (> 10(-5) M) caused a rapid cell death by necrosis. The cell cycle position of apoptotic cells was assessed by the terminal deoxynucleotidyl transferase (TdT) assay of DNA strand breaks combined with DNA staining. MTX, Ara-C, CPT, VP-16 and ADR preferentially induced apoptosis in the S phase. PRD and 6MP induced apoptosis in the G1 phase and G1 + S phases, respectively. CPM showed no cell cycle phase specificity. These findings suggested that the susceptibility of cells to apoptosis was not the sole determinant of cellular sensitivity of cytotoxic drugs. PMID- 9220664 TI - Treatment outcome of AT-B88 regimen for B-cell non-Hodgkin's lymphoma and surface immunoglobulin-positive acute lymphoblastic leukemia in children. AB - We conducted a multicenter study to improve the treatment of B-cell non-Hodgkin's lymphoma and acute lymphoblastic leukemia (NHL/ALL) in Japanese children. The subjects were a total of 57 untreated patients with the B-cell type of either NHL (27 Burkitt's and 9 diffuse large) or ALL between 2 and 15 years old (median: 8 years) seen between 1988 and 1994. All patients received the same cytoreductive therapy (half doses of vincristine and prednisolone) and the same first and second induction courses (vincristine, prednisolone, high-dose methotrexate, repetitive high-dose cyclophosphamide, and adriamycin). Three cycles of consolidation blocks A and B (consisting of reduced doses of similar agents used in the second induction course) followed for patients with stage III or IV NHL or B-ALL, while only one cycle was given for stage I or II disease. Fifty-three patients (93.0%) achieved complete remission. Eight patients had relapse all occurring within 1 year. Another patient had secondary myelodysplastic syndrome. The median follow-up period was 48 months (range: 24-94 months). The overall survival and event-free survival (EFS) rates for all patients were, respectively, 75.9% (S.E.: 5.9) and 70.1 (S.E.: 6.1). The relapse-free interval rate of the 53 patients who achieved CR was 83.5% (S.E.: 5.3). EFS was 75.0% (S.E.: 21.7) in stage I, 84.6% (S.E.: 10.0) in stage II, 78.63% (S.E.: 11.0) in stage III, 80.0% (S.E.: 17.9) in stage IV, and 52.4% (S.E.: 10.9) in ALL. Among the stage IV NHL and ALL patients, EFS was significantly worse in patients with initial CNS involvement than in those without it (14.3% (S.E.: 13.2) vs. 73.7% (S.E.: 10.1); P = 0.0025). In conclusion, our regimen (AT-B88) produced a more than 70% cure rate for children with any stage of B-cell NHL/ALL without initial CNS involvement. However, a new regimen is needed for patients with initial CNS involvement. PMID- 9220665 TI - Effects and serum levels of thrombopoietin in a case of chronic thrombocytopenia with achondroplasia. AB - Thrombopoietin (TPO), produced mainly in the liver, is a major regulator of platelet production. Serum TPO levels are generally increased in thrombocytopenia. We report a case of a 12-year-old boy with chronic severe thrombocytopenia, achondroplasia and nephritis. Severe chronic thrombocytopenia was found at 9 months of age. It was resistant to any treatment. Studies on megakaryocytic colonies in vitro revealed that the marrow cells responded well to TPO and no plasma inhibitor was found. Although hepatic function test results were normal, serum TPO levels in the patient (0.94 fmol/ml) were consistent with those in age-matched children (0.49-1.75 fmol/ml). Chronic thrombocytopenia requires individual evaluation before clinical trials with TPO. PMID- 9220666 TI - Treatment outcome and prognostic factors in childhood acute myeloblastic leukemia: a report from the Japanese Children's Cancer and Leukemia Study Group (CCLSG) AB - Treatment outcome and prognostic factors were evaluated in 152 children with acute myeloblastic leukemia (AML) treated on three consecutive protocols (ANLL 861, 8912, 9205) of the Children's Cancer Leukemia Study Group (CCLSG, Japan). In the ANLL 9205 protocol, anthracycline was used with a continuous infusion of cytosine arabinoside, followed by an intensive sequential post remission chemotherapy of short duration. Forty-two of these 46 patients (91.3%) achieved complete remission, and 58.8% of these patients projected a 3-year disease-free survival. These results were apparently superior to those obtained with the ANLL 861 and 8912 protocols, which used conventional doses of multiple drugs followed by a moderate post remission chemotherapy of long duration. This favorable response with the ANLL 9205 protocol was attributed mainly to the high induction rate of patients with the M4 and M5 FAB subtypes, as compared to those in the previous two protocols (93.3% in ANLL 9205 vs. 57.9% in ANLL 861 + 8912; P < 0.05). The ANLL 861 and 8912 protocols, an older age (> or = 8 years), higher WBC counts (> or = 10 x 10(9)/1) and all predicted an increased risk of relapse and decreased the survival following univariate analysis (P < 0.05). An older age and high WBC count continued to predict an increased risk of relapse in multivariate analyses: patients with an age > 8 years and WBC counts > 10 x 10(9)/1 had a 4.5 times higher risk of relapse than patients without these adverse features. PMID- 9220668 TI - Detection of donor-type alveolar macrophages in transplantation with highly purified CD34+ bone marrow cells. PMID- 9220667 TI - Overexpression of PRAD1/cyclin D1 in plasma cell leukemia with t(11;14)(q13;q32). AB - Two patients with plasma cell leukemia (PCL) with a t(11;14)(q13;q32) translocation are reported. Case 1 is a 64-year-old woman diagnosed as having primary PCL (IgA/lambda, Stage III) with high serum LDH and beta 2-microglobulin (beta 2MG) levels. She was treated with combination chemotherapy but died of gastrointestinal bleeding on the 45th hospital day. Case 2 is a 52-year-old man, initially diagnosed with multiple myeloma (IgG/kappa, Stage III) in August 1993. Relapse several months after primary chemotherapy was characterized by a rapid increase in plasma cells in peripheral blood, high serum LDH and beta 2MG levels, and resistance to further chemotherapy. Both cases showed complex karyotypic abnormalities including t(11;14), and Northern analysis revealed overexpression of the PRAD1/ cyclin D1 gene. The PRAD1 gene is found on chromosome band 11q13 and encodes cyclin D1. Cyclin D1 plays an important role in control of the cell cycle, and overexpression of PRAD1/cyclin D1 may be involved in disease progression in these cases. PMID- 9220669 TI - Transmission of the agent of human granulocytic ehrlichiosis by host-seeking Ixodus scapularis (Acari:Ixodidae) in southern New York state. AB - Ixodes scapularis Say nymphs collected from a natural focus of human granulocytic ehrlichiosis (HGE) in Westchester County, New York, transmitted the HGE agent to uninfected mice in the laboratory. Infection was demonstrated in 3 of 8 mice by polymerase chain reaction analysis of whole blood and microscopic examination of blood smears for morulae. Two of these mice were also positive by xenodiagnosis. Positive xenodiagnostic larvae maintained infection through molting and transferred infection to 1 of 3 mice. Naturally infected I. scapularis ticks transmit the agent of human granulocytic ehrlichiosis to mice, but both acquisition and transmission of this agent by I. scapularis appear to be less efficient than would be expected for Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, the agent of Lyme disease. PMID- 9220670 TI - Morphometric separation of females of Phlebotomus (Phlebotomus) duboscqi and P. (P.) bergeroti (Diptera:Psychodidae). AB - Morphometric evaluation of 27 characters of allopatric females of the closely related species, Phlebotomus duboscqi Neveu-Lemaire and P. bergeroti Parrot, was made in Ethiopia with the aim of finding reliable means to distinguish sympatric specimens. By applying stepwise discriminant analysis, 100% correct classification was obtained using 8 characters. However, 2 of these characters, C3 and C4 (respective distances from sockets of the longest ascoids on segment 3 and 4 to distal margin of the segment) with the highest discriminant loadings, separated these 2 species with a success rate of 98%. It is recommended that these 2 characters be used in the routine identification of these species. PMID- 9220671 TI - Density of larval Culicoides belkini (Diptera:Ceratopogonidae) in relation to physicochemical variables in different habitats. AB - Immature density and population size of the biting midge Culicoides belkini (Wirth & Arnaud) were estimated for habitats on Moorea Island, French Polynesia, by means of random, 2- and 3-stage sampling designs. Samples were taken in March 1993 from 5 strata of a large larval habitat: a sandy-mud surface of approximately 5,000 m2 (stratum 1) in which approximately 12,000 land crab burrows (stratum 2) were counted, a small pond surrounded by approximately 300 m2 of muddy bank (stratum 3), and a high organic muddy area (Kopara) of approximately 1,200 m2 (stratum 4) with approximately 3,500 crab burrows (stratum 5). Larval density was usually higher in the mud of crab burrows, especially those in the Kopara stratum. Larval density was significantly lower in the sediment of the sandy area as compared with pond banks or Kopara surface. The sampling designs and techniques were logistically adequate, statistically relevant, and were recommended for future studies on C. belkini larval density. Larval habitats were characterized by means of multivariate analysis. Comparison of larval densities with selected environmental variables indicated that larvae density was higher in wet sediments with high levels of organic matter (approximately 8% of dry weight of sediment) and low salinity (approximately 0.5 1.5% NaCl equivalents). These variables were considered significant if larval control by means of habitat modification has to be achieved. Nevertheless, C. belkini can tolerate a broad spectrum of variation in the other environmental variables measured and breed in a variety of ecological situations. Therefore, it has a high potential for colonizing new habitats. PMID- 9220672 TI - Comparison of behavior and vector efficiency of Anopheles gambiae and An. arabiensis (Diptera:Culicidae) in Barkedji, a Sahelian area of Senegal. AB - The ecology, population dynamics, and malaria vector efficiency of Anopheles gambiae and An. arabiensis were studied for 2 yr in a Sahelian village of Senegal. Anophelines were captured at human bait and resting indoors by pyrethrum spray. Mosquitoes belonging to the An. gambiae complex were identified by polymerase chain reaction. Of 26,973 females, An. arabiensis represented 79% of the mosquitoes captured and remained in the study area longer than An. gambiae after the rains terminated. There were no differences in nocturnal biting cycles or endophagous rates between An. gambiae and An. arabiensis. Based on an enzyme linked immunosorbent assay test of bloodmeals, the anthropophilic rate of these 2 vectors were both approximately 60%, when comparisons were made during the same period. Overall, 18% of the resting females had patent mixed bloodmeals, mainly human-bovine. The parity rates of An. gambiae and An. arabiensis varied temporally. Despite similar behavior, the Plasmodium falciparum circumsporozoite protein (CSP) rates were different between An. gambiae (4.1%) and An. arabiensis (1.3%). P. malariae and P. ovale only represented 4% of the total Plasmodium identified in mosquitoes. Transmission was seasonal, occurring mainly during 4 mo. The CSP entomological inoculation rates were 128 bites per human per year for the 1st yr and 100 for the 2nd yr. Because of the combination of a high human biting rate and a low CSP rate, An. arabiensis accounted for 63% of transmission. Possible origin of differences in CSP rate between An. gambiae and An. arabiensis is discussed in relation to the parity rate, blood feeding frequency, and the hypothesis of genetic factors. PMID- 9220673 TI - Regional molecular genetic key of thirteen snow pool Aedes species (Diptera:Culicidae) in northern Colorado. AB - Snow pool Aedes mosquitoes are identified easily and accurately by microscopic examination of 4th instars or male genitalia. Early instars and females cannot be identified accurately. We demonstrate that restriction fragment-length polymorphisms (RFLP) in the internal transcribed spacer (ITS) of the nuclear ribosomal DNA cistron are specific to each of 13 snow pool Aedes species found in northern Colorado. The ITS was amplified by the polymerase chain reaction (PCR) and digested with AluI and MspI restriction endonucleases. Differences in the sizes of digested fragments among the 13 species were so slight that they could only be resolved with polyacrylamide gel electrophoresis and visualized with silver staining. A key to species found in the Rocky Mountains of northern Colorado was constructed using the PCR-RFLP patterns. Three of the species were collected in California and had identical PCR-RFLP patterns, indicating that restriction sites in the ITS may be conserved within some species. PMID- 9220674 TI - New records of subcutaneous mites (Acari:Hypoderatidae) in birds, with examples of potential host colonization events. AB - New host, geographic records, or both are established for 14 species of hypoderatid deutonymphs from 14 species of birds in North America. Ten of these records are regarded as examples of a potential host colonization event where these hypopi have become established in hosts other than those with which they are normally associated. Herein, potential host colonization events by hypoderatid deutonymphs are regarded as more of an ecologically determined than physiologically specific phenomenon, often specifically related to sharing of nesting sites in the same rookeries by different host taxa. Neottialges ibisicola Young & Pence is placed as a junior synonym of Neottialges plegadicola Fain. The taxonomic status of Hypodectes propus from columbid versus ardeid hosts needs further study. PMID- 9220676 TI - Importance of supercooling points in the overwintering of the horn fly and stable fly (Diptera:Muscidae). AB - Supercooling points were determined for eggs, 3rd instars, pupae, newly emerged unfed adults and 3-d-old engorged laboratory reared adults of Haematobia irritans (L.) and Stomoxys calcitrans (L.). Wild nondiapausing and diapausing pupae of H. irritans also were tested. Mean supercooling points ranged from -28.0 degrees C for H. irritans eggs to -6.8 degrees C for H. irritans larvae. Mean supercooling points of all H. irritans developmental stages were lower than those of comparable S. calcitrans developmental stages, with the exception of larvae where the reverse was true. The mean supercooling point of diapausing H. irritans pupae (-23.5 degrees C) was significantly lower than those of nondiapausing laboratory pupae (-20.8 degrees C) or nondiapausing wild pupae (-20.2 degrees C). Developmental stages of both species were freeze intolerant, with no survival following exposures to temperatures below the supercooling points. Results are discussed with respect to the disparate overwintering strategies of these species and in relation to typical climatic minima experienced in south central Texas. The cold tolerance of H. irritans and S. calcitrans pupae was compared at 4 degrees C, a temperature below their developmental threshold of 11.5 degrees C and above their mean supercooling points. The survival of H. irritans pupae was significantly greater than the survival of S. calcitrans pupae. Cold injury was a significant mortality factor for both species. PMID- 9220677 TI - Time of host-seeking by Culex tarsalis (Diptera:Culicidae) in California. AB - Factors altering the pattern of Culex tarsalis Coquillett host-seeking activity were studied in Kern and Riverside counties of California using an automatic time segregated sampler baited with bottled CO2 gas released at 0.5 or 1.0 liters/min. Host-seeking always commenced shortly after sunset and usually peaked during the succeeding 1-3 h, the hottest and driest time of the night. The time of maximal activity varied over time and space, because of increased mosquito abundance (presumably reduced blood feeding success), distance of the sampler from resting sites, adulticide applications, and perhaps weather. PMID- 9220678 TI - The genus Hexidionis (Acari:Trombiculidae) with the description of a new species from Texas. AB - Hexidionis garfieldi is described as new from specimens collected off a domestic cat. Felis silvestris f. catus Schreber, in Corpus Christi, TX. The genus Hexidionis Vercammen-Grandjean & Loomis, 1967, is rediagnosed and a key to the 11 included species given. PMID- 9220675 TI - Mixed-species Plasmodium infections of Anopheles (Diptera:Culicidae) AB - Mixed-pathogen infections of vectors rarely are considered in the epidemiological literature, although they may occur in nature. A review of published reports shows that many Anopheles species are capable of carrying sporozoites of > 1 Plasmodium species, of doing so simultaneously in field conditions, and of acquiring and transmitting these in experimental situations. Mixed-species infections in mosquito populations occur at frequencies greater than or equal to the product of the constituent species prevalences, whereas human populations have apparent mixed-species infections at frequencies less than or equal to their corresponding expected values. We present a model for the accumulation of parasite infections over the lifespan of a mosquito that explains this surplus of mixed-species infections. However, the expected frequencies of mixed infections on the basis of our model are greater than those found in nature, indicating that the sampling by mosquitoes of Plasmodium species from human malaria infections may not be random. PMID- 9220679 TI - Population genetic structure of Ixodes pacificus (Acari:Ixodidae) using allozymes. AB - Genetic analysis of the population structure of the western blacklegged tick, Ixodes pacificus Cooley & Kohls, was conducted using allozymes. This vector tick transmits the Lyme disease spirochete, Borrelia burgdorferi Johnson, Schmid, Hyde, Steigerwalt & Brenner, in the far-western United States. It ranges from British Columbia to Baja California and disjunct populations are present in Oregon, Nevada, Utah, and Arizona. Host-seeking adult ticks were collected from vegetation across the range of the species and were partially fed on rabbits prior to analysis. Twelve putative loci were resolved using starch gel electrophoresis. One locus, glucose-6-phosphate isomerase, formed an apparent north/south latitudinal cline and showed significant geographic structure. None of the remaining loci exhibited much genetic differentiation. Estimates of gene flow were high relative to other arthropods. Isolation-by-distance analysis suggests a recent and rapid range expansion. We conclude that the overall lack of differentiation is due high rates of gene flow. PMID- 9220681 TI - Effects of developmental asynchrony between Aedes triseriatus (Diptera:Culicidae) and its predator Toxorhynchites rutilus (Diptera:Culicidae). AB - Newly hatched Toxorhynchites rutilus (Coquillet) were added to experimental populations of Aedes triseriatus (Say) at varying days after prey hatch to test the hypothesis that a developmental asynchrony of Ae. triseriatus and Tx. rutilus leads to escape from predation by Ae. triseriatus in small water bodies. Presence of Tx. rutilus significantly affected prey survivorship. Regression of survivorship [log10 (x + 1) transformed] versus days head start for prey yielded a small, but significant positive slope, indicating that survivorship increased slightly with an increasing number of days head start. For females, mean weight at emergence was not significantly affected by treatments; however, median days to emergence differed significantly between the treatments, with females taking significantly longer to emerge with Tx. rutilus absent than when the predator was present. For males, neither mean mass nor median days to emergence was significantly affected by treatments. Treatments had no significant effect on the frequency of deaths or on mean weight of Tx. rutilus. Thus, a developmental asynchrony between Tx. rutilus and Ae. Triseriatus appears to have no effects on the predator, but does have a weak effect on prey performance at high hatch densities. PMID- 9220680 TI - Ability of the Lyme disease spirochete Borrelia burgdorferi to infect rodents and three species of human-biting ticks (blacklegged tick, American dog tick, lone star tick) (Acari:Ixodidae). AB - The infectivity of a diverse collection of Borrelia burgdorferi strains from North America for mice was determined as a prelude to vector competence experiments with the 3 primary human-biting tick species in the eastern United States (Ixodes scapularis Say, Dermacentor variabilis (Say), Amblyomma americanum (L.)]. Of the 34 B. burgdorferi strains inoculated into mice, 29 were infectious; the exceptions were 5 isolates from Texas. Vector competence experiments were conducted with 2 strains from the southern United States (North Carolina and Georgia). Both strains were extremely infectious to I. scapularis larvae. Moreover, I. scapularis efficiently maintained these spirochetes transstadially and transmitted infection as nymphs. D. variabilis larvae were intermediate in susceptibility but generally did not maintain the infection transstadially. A. americanum larvae were completely refractory to infection with these 2 southern B. burgdorferi strains. Three isolates from Michigan D. variabilis were inoculated into mice, subsequently exposed to I. scapularis and D. variabilis larvae. Larval I. scapularis were 5-fold more susceptible to infection with these strains than were larval D. variabilis. Although nymphal I. scapularis efficiently transmitted a Michigan isolate, nymphal D. variabilis did not. In all these experiments, I. scapularis was the only species that proved to be vector competent for B. burgdorferi. PMID- 9220682 TI - Simulation of blacklegged tick (Acari:Ixodidae) population dynamics and transmission of Borrelia burgdorferi. AB - A model (LYMESIM) was developed for computer simulation of blacklegged tick, Ixodes scapularis Say, population dynamics and transmission of the Lyme disease agent. Borrelia burgdorferi Johnson. Schmid, Hyde, Steigerwalt & Brenner, LYMESIM simulates the effects of ambient temperature, saturation deficit, precipitation, habitat type, and host type and density on tick populations. Epidemiological parameters including host infectivity, tick infectivity, transovarial transmission, and transstadial transmission are included in the model to simulate transmission of the Lyme disease spirochete between vector ticks and vertebrate hosts. Validity of LYMESIM was established by comparing simulated and observed populations of immature I. scapularis on white-footed mice. Peromyscus leucopus, (Rafinesque), at 2 locations in Massachusetts. Validity also was indicated by comparisons of simulated and observed seasonality of blacklegged ticks in New York, Massachusetts, Florida, and Oklahoma-Arkansas. Further model validity was shown by correlation between simulated and observed numbers of immature ticks engorging on white-footed mice at 3 sites in Massachusetts. The model produced acceptable values for initial population growth rate, generation time, and 20-yr population density when historical meteorological data for 16 locations in eastern North America were used. Realistic rates of infection in ticks were produced for locations in the northeastern and northcentral United States. LYMESIM was used to study the effect of white-footed mouse and white-tailed deer, Odocoileus virginianus (Zimmerman), densities on tick density and infection rates. The model was also used to estimate tick density thresholds for maintenance of B. burgdorferi. PMID- 9220683 TI - Effect of juvenile hormone and juvenile hormone mimics on sperm transfer from the testes of the male cat flea (Siphonaptera:Pulicidae). AB - Sperm transfer into the epididymis was completed without a blood meal, when newly emerged male cat fleas. Ctenocephalides felis (Bouche), were exposed to filter papers treated with juvenile hormone III or the juvenile hormone mimics fenoxycarb, methoprene, or pyriproxyfen. As the concentration of juvenile hormone or the time of flea exposure to juvenile hormone or the juvenile hormone mimics increased, the percentage of fleas that transferred sperm also increased. The percentage of pyriproxyfen-treated males that transferred sperm reached 100% after 3 d: whereas, 7 d exposure to juvenile hormone, fenoxycarb and methoprene was required for 100% of the males to transfer sperm. Although sperm were present in the epididymis of treated fleas, insemination of females did not take place off the host either on juvenile hormone-treated filter paper or on juvenile hormone-treated dog hair. PMID- 9220684 TI - Competence of urban rats as reservoir hosts for Lyme disease spirochetes. AB - To determine whether urban rats serve efficiently as reservoir hosts for the agent of Lyme disease, we recorded the frequency of infection in nymphal Ixodes ricinus (L.) ticks that had fed as larvae on experimentally infected Norway rats, Rattus norvegicus (Berkenhout), or on black rats, R. rattus (L.), and evaluated the nidicolous venue of transmission. Subadult vector ticks attached readily to Norway rats as well as black rats and virtually all became infected in the course of feeding. Larval ticks detached when these nocturnally active hosts were at rest. Rats appeared to be competent reservoir hosts of Lyme disease spirochetes in a transmission cycle in urban sites. PMID- 9220685 TI - Effects of larval nutrition on the postembryonic development of Ctenocephalides felis felis (Siphonaptera:Pulicidae). AB - Dry blood from mammals and birds was used as larval diet for the development of the cat flea, Ctenocephalides felis felis (Bouche), in the laboratory. Diets that contained host blood and cornmeal heated at 40 degrees C for 30 min were inadequate for most larvae to form pupae. Development time from 1st instar to adult ranged from 30 to 33 d. Except for the diet containing Mastomys blood, the diets that consisted of blood alone from other hosts air dried at room temperature contained sufficient nutritional value to allow adults to be obtained from > 51.7% of larvae fed these diets. Adults were obtained from > 81% of pupae. Although the Mastomys or mouse blood contributed to better diets than the dog or pigeon blood through shorter developmental time from 1st instar to adults, greater numbers of pupae and adults were obtained from diets that contained dog and mouse blood. Highly significant differences existed between pigeon and Mastomys blood in relation to the number of cocoons formed and between pigeon and dog blood or pigeon and mouse blood in relation to adult emergence. Differences between dog and mouse blood in the larval diet were significant only in relation to mortality that occurred to the pupae. PMID- 9220687 TI - Presentation of Mississippi Insurance Commissioner George Dale MSMA 129th annual session. PMID- 9220686 TI - Brain metastases from unknown primary site. AB - We report our experience with 32 patients presenting with brain metastasis from unknown origin. This constitutes 11.5 percent of 276 consecutive patients with brain metastasis seen over a period of eight years at the University of Mississippi Medical Center. Patients with solitary resectable lesion underwent surgery followed by irradiation. All patients with multiple metastasis were treated with whole brain radiation and dexarnethasone. The mean survival was 31.50 weeks for patients with single lesion and 19.11 weeks for multiple lesions. The investigations and treatment management of brain metastasis from unknown primary site are discussed. PMID- 9220688 TI - Address of the president MSMA House of Delegates--May 15, 1997. PMID- 9220689 TI - Medicaid patients access specialists. PMID- 9220690 TI - HCQIP Project Report--findings released in Use of Ace Inhibitors in Heart Failure Project. PMID- 9220691 TI - Pancreatitis associated with measles in a young adult. PMID- 9220692 TI - Report of the NMA panel on mammography. PMID- 9220693 TI - Managed care, maniac care, rationed care, or no care: does anyone care? PMID- 9220694 TI - Cost containment for treating hypertension in African Americans: impact of a combined ACE inhibitor-calcium channel blocker. AB - The use of calcium channel blockers (CCBs) and angiotensin-converting enzyme (ACE) inhibitors has increased dramatically over the last 10 years and now accounts for 60% to 70% of all new antihypertensive prescriptions. Even though these two classes are efficacious, they are costly. Combined ACE inhibitor/CCB therapy (amlodipine-benazepril) was introduced in 1995. An analysis was done to assess the potential financial impact of substituting this agent for patients being treated with on ACE inhibitor/CCB combination. A pharmaceutical profile review of prescriptions during October 1995 was performed on 219 randomly selected patients enrolled in a Medicaid managed care program. Eighty-four profiles were analyzed; 24% of patients were on a combination ACE inhibitor/CCB regimen with an average monthly cost of $135. If the single agent amlodipine benazepril with an average monthly cost of $45 (all strengths) was substituted, the savings would be considerable: $1080 per patient per year and $1,080,000 annualized for the calculated number of hypertensives on combination therapy in our network of 15,000 patients. Therapeutic substitution is one method of achieving cost containment in managed care. The cost differential between separately prescribed CCBs and ACE inhibitors and amlodipine-benazepril is significant. Compliance also should be enhanced as the patient would need to take only one pill daily. Once a patient has been maintained on a stable dose of a CCB/ACE inhibitor, substitution with amlodipine-benazepril should be considered. PMID- 9220695 TI - Schistosomiasis: an unusual cause of right lower quadrant abdominal pain. AB - Schistosomiasis, although unusual in North America, is a common disease worldwide. Symptoms vary depending on the species involved. Immigrants from endemic regions are the commonly affected patients found in North America. In most cases, schistosomiasis does not present with right lower quadrant pain. Even in endemic regions, this form of presentation is uncommon. In the United States, most cases of right lower quadrant pain often will be treated as appendicitis. Questions remain unanswered as to whether the schistosomes cause appendicitis or are found incidentally in these cases. Stool and urine specimens may be helpful in making a diagnosis. Most cases require operative intervention to rule out appendicitis and to obtain tissue for histopathologic diagnosis. Praziquantel is effective in eradicating infestations. PMID- 9220697 TI - The application of meta-analysis in assessing racial differences in the effects of antihypertensive medication. AB - Meta-analysis is an important technique for synthesizing research findings. Although the statistical foundations of meta-analysis continue to be debated, few question its value as a rigorous framework for organizing literature reviews. In recent years, there has been increasing emphasis on the use of meta-analysis not only to summarize the central tendency of findings but also to explain variation between studies. This article reviews the major steps in a meta-analysis with an emphasis on comparative analysis of subgroups of studies. A meta-analysis of the antihypertensive efficacy of calcium channel blockers is used to illustrate how a comparative analysis can be applied to investigate racial variation in the effects of calcium channel blockers. A statistically significant trend is found between the proportion of African-American hypertensive subjects and the mean reduction in blood pressure. Meta-analytic techniques also are applied to explore possible confounders due to differences in research design and patient characteristics. PMID- 9220698 TI - Homage to the NMA: the NDA story (1895 to 1975)--Part 1. PMID- 9220696 TI - Stress, stress reduction, and hypertension in African Americans: an updated review. AB - This is a comprehensive and integrative review of multiple factors underlying the greater prevalence of hypertension in African Americans compared with whites. Evidence linking stress with hypertension and cardiovascular disease in African Americans is reviewed. A survey of mechanisms of hypertension in African Americans and existing behavioral strategies for the treatment of hypertension is presented. Given that the excess of hypertension may be mediated in part by behavioral factors operating through biological mechanisms, a case is presented for behavioral stress reduction measures. This review of stress reduction techniques especially the Transcendental Mediation program for the treatment of hypertension in African Americans highlights current issues facing the field. New information is provided to help direct future nonpharmacological research and practice in hypertension to prevent morbidity and premature mortality in this underserved population. PMID- 9220699 TI - HIV risk perception, risk behavior, and seroprevalence among female commercial sex workers in Georgetown, Guyana. AB - A study of 108 female sex workers engaged in prostitution in Georgetown, Guyana, was made in April 1993. Based on interviews and procurement of blood samples, the study investigated relationships between HIV seroprevalences and AIDS knowledge, risk behaviors, client characteristics, and condom use. Street-walkers-as distinct from sex workers in bars, hotels, and Port Georgetown-tended to charge less, be worse off socioeconomically, and have clients who were similarly disadvantaged; they were therefore classified as belonging to a "lower" socio economic stratum, while the other workers were classified as belonging to a "higher" stratum. The overall HIV seroprevalence found among the sex workers was 25% (95% CI: 17%-33%). But the 50 subjects in the lower stratum had a relatively high seroprevalence (42%, as compared to 10% among those in the higher stratum), accounting for 21 of the 27 HIV-seropositive subjects. Reported patterns of client origins (Guyanese or foreign), worker willingness to have sex without a condom, and condom use by clients differed by stratum. Participants in the higher stratum were more disposed to having sex without a condom. The workers' knowledge of what causes AIDS and how HIV is transmitted was low in both strata; substantial numbers of workers said they had contracted a sexually transmitted disease within the past two years or were users of illicit drugs. Condom use is reportedly less common among Guyanese than foreign clients, suggesting a greater risk of contracting HIV from Guyanese clients or infecting Guyanese clients with it. The HIV seroprevalence among workers who said they had only Guyanese clients was statistically greater than the rate among those who said they had only foreign clients. The HIV seroprevalence among those reporting more than five clients per week was statistically greater than among those reporting fewer. HIV seropositivity was relatively high among the 12 workers who said they used cocaine. Overall, the findings supported the view that interventions targeted at female sex workers and their clients should be strengthened-more specifically, that concerted efforts should be made to intensity condom promotion, distribution, and social marketing; to improve STD services that provide treatment and counseling for female sex workers; and to increase educational activities among the workers' Guyanese clients. PMID- 9220700 TI - Commerce in health services in North America within the context of the North American Free Trade Agreement. AB - This article discusses the future of commercial trade in personal health services in North America within the context of the North American Free Trade Agreement (NAFTA) and the latter's potential influence on health care for the Mexican people. It begins by defining concepts related to international trade of services, particularly health services, and then proceeds to analyze elements of NAFTA that affect the delivery, regulation, and financing of such services, as well as their future trade within the NAFTA area. It concludes with some recommendations directed at helping Mexico's national health care system confront the risks posed while taking advantage of the opportunities offered by the Mexican economy's entry into a broader market. PMID- 9220701 TI - Expanded programme on immunization (EPI). Progress towards the global eradication of poliomyelitis, 1996. PMID- 9220702 TI - Zoonoses control. Lyssavirus infection in 3 fruit bats, Australia. PMID- 9220703 TI - Two-year group treatment for children with learning difficulties: assessing effects of treatment duration and pretreatment characteristics. AB - The results of a 2-year treatment study of children with learning problems are reported. During the first treatment year, half of the children participated in a multifaceted neurocognitive treatment and the other half in a treatment that provided supervision of school tasks and peer group support. During the second treatment year, all children participated in the neurocognitive treatment. The participants were 74 Chilean children 6 to 11 years old. The issues under investigation were the effect of treatment duration, and the relationship between pretreatment neurocognitive and behavioral characteristics and academic treatment outcome. The results indicated that significant gains occurred during both the first and the second treatment year. No major differences were found between the treatment groups. Pretreatment negative behavioral traits were associated with lesser academic growth in the group participating in the homework supervision treatment but not in the neurocognitive treatment group. PMID- 9220704 TI - Phonologic impairment and prereading: update on a longitudinal study. AB - This study examined the effects of overt phonologic impairment (disordered speech) on phonological awareness, verbal working memory, and letter knowledge. Forty-five children--29 with moderate to severe productive phonologic impairment at the inception of the project and 16 without impairment--were followed from mean age 3-6 to age 6-0. Fifteen participants with impairment were matched on gender and mental age to 15 without impairment for certain aspects of the analysis. The children with phonologic impairment performed significantly worse than their controls on tasks of verbal working memory, phoneme segmentation, and letter identification. In addition, a path analysis revealed working memory to be a potentially important mediating variable. The investigators also measured productive syntax, which, although associated with productive phonology and working memory, was not associated with letter identification. PMID- 9220706 TI - Re-inventing government? Let's re-invent special education. AB - Persistent problems in meeting the intent of Public Law 94-142 (and its reauthorization legislation) are caused by the failure of federal and state leaders to collaborate beyond the boundaries of current policies to develop new approaches to meeting the learning needs of students with disabilities. Much of the hindrance derives from problems with current referral-to-placement procedures. This article proposes a way to escape from the quagmire of laws, regulations, and policies that make special education costly rather than free and that inhibit teachers from developing appropriate educational strategies. PMID- 9220707 TI - Key questions related to building collaborative and inclusive schools. AB - In this article the author provides the reader with 15 key questions, the answers to which are designed to facilitate the development of collaborative and inclusive schools. The questions are organized into three categories: general and philosophical questions pertaining to inclusion, some questions about the basic mechanics of developing inclusion programs, and some questions about the practical implementation of inclusion. Emphasis is placed on the provision of a modified continuum of services via a variety of ways of supporting teachers in inclusive classrooms. PMID- 9220708 TI - Co-teaching experiences: the benefits and problems that teachers and principals report over time. AB - This report describes a 3-year study of 18 elementary and 7 middle school teams involved in the development and implementation of building-level programs designed to support students with disabilities in mainstream classrooms. All of the teams used co-teaching as an integral part of their service delivery models. During the investigation, 119 teachers and 24 administrators participated in 1 or more years of data collection. The emerging benefits and persistent problems that participants identified in the development of their models are described. PMID- 9220709 TI - The BRIDGE project: bridging the gap between university and schools. Being Responsive to Individual Differences in General Education. AB - BRIDGE (Being Responsive to Individual Differences in General Education) is a partnership between university faculty and two elementary schools, focusing on the collaborative practices and programs in general and special education. University faculty have adopted new roles as facilitators, rather than "experts," in working side by side with teachers seeking to increase their sense of professional efficacy. Teachers at the two sites collaborate with university faculty and each other in acquiring new information and practices within four initiative areas. The four initiatives, designed generally to support professional development toward meeting the needs of struggling students, are (a) collaboration between special and general education, (b) family liaisons, (c) teacher research, and (d) teacher education. University student teachers at these two schools receive their clinical education in settings ripe with teacher inquiry, collaboration, and ongoing development of new and creative strategies. PMID- 9220711 TI - Meeting the challenge of consultation and collaboration: developing interactive teams. AB - The roles of special educators as consultants and collaborators have long been established and supported. The rationale for these roles is also well documented. Many models--consultative, collaborative, and teaming--have been suggested in the literature; sometimes, these models exhibit similar goals, competencies, and processes. Because of intensified pressures to collaborate, successful implementation of collaborative and team efforts requires that special educators expand their roles as interactive professionals. The purpose of this article is to define and describe the consultation, collaboration, and teaming models that have been implemented, discuss their strengths and limitations, delineate how these models contribute to interactive teaming, outline key features of the interactive team, and provide some guiding principles for successful implementation. PMID- 9220710 TI - Collaborative speech and language services for students with learning disabilities. AB - Fueled by educational reforms such as the Regular Education Initiative, the inclusion movement, and Goals 2000, speech and language pathologists (SLPs) have explored the use of collaborative consultation in providing integrated service delivery. The implications of classroom-based services are discussed, along with models that have been adopted by SLPs, learning disabilities specialists (LDSs), and classroom teachers. The characteristics of students served, and the areas of speech and language (i.e., language, articulation, fluency, voice) targeted in the classroom, are reviewed. Ways in which SLPs, LDSs, and classroom teachers can collaborate, including collaborative assessment; Individualized Education Program development; teaching listening, speaking, reading, and writing skills; and teaching students the language of the classroom, are described. PMID- 9220712 TI - Renovating and refurbishing the field experience structures for novice teachers. AB - Teacher education at both the preservice and the inservice level has been criticized for not adequately preparing teachers for the demands and challenges in the classrooms of today, or for the future. This article describes a federally funded project awarded through the Office of Special Education Programs. The intended outcome was to improve the knowledge and application of professional skills and competencies for both preservice teachers (student teachers) and inservice teachers (supervising master teachers) through continuous, focused staff development, reflection, discussion, and coaching. The project focused attention on three related issues: (a) the role and training of the supervising and/or master teacher, (b) better partnerships between teacher preparation programs and the public schools, and (c) the effects of school structure on teachers' continued professional growth. This article describes the program planning, goals, activities, components, and impact of this personnel preparation project. PMID- 9220713 TI - If it takes two to tango, then why not teach both partners to dance? Collaboration instruction for all educators. AB - Being able to collaborate effectively is important for teachers who work together to serve students with learning disabilities in general education classrooms. Effective collaboration requires that teachers have knowledge and skills in how to effectively communicate and share their technical expertise for the purpose of solving classroom problems and providing continuity across instructional settings. Although both special education and general education preparation programs provide preservice teachers with the technical expertise for their respective areas of certification, few programs provide both special education and general education majors with instruction in interpersonal communication skills and collaboration strategies. The purpose of this article is to suggest guidelines and strategies to help teacher preparation programs move toward collaboration instruction for all educators. Suggestions for what to teach and how to teach it are offered, as well as an overview of factors that influence the implementation of collaboration instruction for all educators. PMID- 9220714 TI - Collaboration and school reform: a twenty-first-century perspective. AB - Collaboration and school reform are current social-political issues that affect many individuals and school districts. In this series summary, the authors examine national goals, federal legislation, and local reform practices. A twenty first-century perspective predicting the integration of collaboration and school reform may help to guide future teaching, research, and/or service efforts. How families and professionals work together today will affect how we educate students during the next century. PMID- 9220715 TI - Psychosocial aspects of patient counseling and selection: a surgeon's perspective. PMID- 9220716 TI - Psychosocial factors in facial surgery: a practical guide. PMID- 9220717 TI - Does facial plastic surgery alter the social perception of patients by others: a cross-national perspective. PMID- 9220718 TI - Psychologic and social consequences of craniofacial disfigurement in children. PMID- 9220719 TI - Psychologic aspects of facial trauma and scarring. PMID- 9220720 TI - The aging face: a psychocutaneous perspective. PMID- 9220721 TI - Psychologic aspects of major head and neck reconstructive surgery. PMID- 9220722 TI - Management of patient dissatisfaction with cosmetic surgery. PMID- 9220723 TI - Medicolegal aspects of otolaryngologic, facial plastic, and reconstructive surgery. PMID- 9220724 TI - Ethics and integrity in facial plastic surgery: imperatives for the 21st century. PMID- 9220725 TI - The aging face. AB - Aging is an inevitable process. Most patients identify the concept of self-image in their facial appearance. The face is also our interface with the outside world. Changes in the face secondary to aging are the most apparent. Some aspects of aging are fairly uncontrollable, and these are largely based on hereditary factors. Other factors are somewhat controllable and are largely the result of exposure to the elements and harmful habits. The processes of facial aging are slowly becoming understood. Although different patients appear to age at different rates, biologic aging does not seem necessarily to follow chronologic aging in the same fashion in different individuals. However, a fundamental pattern as well as sequence in aging is fairly predictable and describable. The effects of ultraviolet radiation and involutional changes are manifested in the skin as wrinkles. Involutional changes in the suspensory structures in the face result in skin laxity and dependency. In this article, the various factors involved in aging are reviewed. The effects of these factors in different regions are discussed, with representative illustrations to represent concepts. PMID- 9220726 TI - Combination medium-depth peeling: the Jessner's + TCA peel. AB - The Jessner's + TCA peel is an enhanced medium-depth peel that is a combination of two acidic compounds. It has been found to be effective as a peeling procedure for moderately photoaging skin, actinic keratoses, and superficial acne scars. PMID- 9220727 TI - Rejuvenation of the skin surface: chemical peel and dermabrasion. AB - Chemical peel and dermabrasion are traditional, well-proven methods for the rejuvenation of the skin. The medium-depth trichloroacetic acid peel and the deep phenol peel offer distinct advantages and disadvantages and are discussed in detail in this article. The management of complications associated with both peel techniques is also discussed. Regional dermabrasion is an effective adjunct to facial rejuvenative surgery, such as face lift and blepharoplasty. Full-face dermabrasion and spot or local dermabrasion are most often used in the treatment of facial scarring. The technique of dermabrasion is discussed as well as its indications and postoperative care. Results are shown for both dermabrasion and peel. PMID- 9220728 TI - Rejuvenation of the skin surface: laser exfoliation. PMID- 9220729 TI - Rejuvenation of the aging forehead and brow. AB - Rejuvenation of the aging upper face can transform tired and angry features into youthful-appearing ones. This article presents the principles for analyzing and treating the aging forehead and brow. The esthetic dimensions and proportions of the brow and forehead are discussed, in context with the corresponding surgical anatomy. The goals of facial rejuvenation surgery as it relates to the upper third of the face are addressed. Various approaches, including their advantages and disadvantages, are presented. The appropriate approach is selected to eliminate unsightly features that are in need of correction while minimizing hairline shifts and forehead scarring and anesthesia. Following the principles and techniques illustrated in this article, the facial plastic surgeon may confidently treat the signs and complaints of the aging upper face. PMID- 9220730 TI - Upper-lid blepharoplasty. PMID- 9220731 TI - Transcutaneous lower blepharoplasty. AB - Transcutaneous lower blepharoplasty as a standard procedure for cosmetic lower blepharoplasty is being replaced by transconjunctival blepharoplasty and laser resurfacing techniques. Nevertheless, there remain specific indications for the transcutaneous approach and its modifications. The accomplished facial plastic surgeon will be able to choose the appropriate technique based on the patient's concerns, the eyelid pathology, and his or her clinical experience. PMID- 9220732 TI - Transconjunctival blepharoplasty. AB - Cosmetic blepharoplasty is directed at the surgical correction of undesirable changes of the eyelids that are usually of an acquired nature and caused by aging. The goals are to improve the appearance and, many times, the function of the eyelids. Just as important as attaining these goals is avoiding any complications or undesirable sequelae of a blepharoplasty procedure. In this article, the technique and application of transconjunctival blepharoplasty are reviewed. The differences in the preseptal and retroseptal approaches are discussed and illustrated. The transconjunctival technique can be expanded through the use of various other techniques in order to apply it to a wider variety of patients. These expanded techniques of transconjunctival blepharoplasty allow the surgeon to manage excess skin and rhytids more effectively. Transconjunctival blepharoplasty, therefore, represents a technique in the armamatarium of surgeons performing cosmetic blepharoplasty that has gained new popularity. The technique can be effectively applied to a wide variety of patients. Transconjunctival blepharoplasty allows the surgeon to accomplish many of the esthetic goals of blepharoplasty while reducing the incidence of many of the associated problems, i.e., lid malposition and a visible cutaneous scar. PMID- 9220733 TI - The aging nose in rhinoplasty for facial rejuvenation. PMID- 9220734 TI - Surgical correction of the aging nose. PMID- 9220735 TI - Cyclic fatigue testing of nickel-titanium endodontic instruments. AB - Cyclic fatigue of nickel-titanium, engine-driven instruments was studied by determining the effect of canal curvature and operating speed on the breakage of Lightspeed instruments. A new method of canal curvature evaluation that addressed both angle and abruptness of curvature was introduced. Canal curvature was simulated by constructing six curved stainless-steel guide tubes with angles of curvature of 30, 45, or 60 degrees, and radii of curvature of 2 or 5 mm. Size #30 and #40 Light-speed instruments were placed through the guide tubes and the heads secured in the collet of a Mangtrol Dynamometer. A simulated operating load of 10 g-cm was applied. Instruments were able to rotate freely in the test apparatus at speeds of 750, 1300, or 2000 rpm until separation occurred. Cycles to failure were determined. Cycles to failure were not affected by rpm. Instruments did not separate at the head, but rather at the point of maximum flexure of the shaft, corresponding to the midpoint of curvature within the guide tube. The instruments with larger diameter shafts, #40, failed after significantly fewer cycles than did #30 instruments under identical test conditions. Multivariable analysis of variance indicated that cycles to failure significantly decreased as the radius of curvature decreased from 5 mm to 2 mm and as the angle of curvature increased greater than 30 degrees (p < 0.05, power = 0.9). Scanning electron microscopic evaluation revealed ductile fracture as the fatigue failure mode. These results indicate that, for nickel-titanium, engine-driven rotary instruments, the radius of curvature, angle of curvature, and instrument size are more important than operating speed for predicting separation. This study supports engineering concepts of cyclic fatigue failure and suggests that standardized fatigue tests of nickel-titanium rotary instruments should include dynamic operation in a flexed state. The results also suggest that the effect of the radius of curvature as an independent variable should be considered when evaluating studies of root canal instrumentation. PMID- 9220737 TI - Analysis of the forces developed during obturation: warm vertical compaction. AB - The aim of this work was to measure and analyze the forces applied by endodontists during an obturation. This was achieved by devising a system of force transducers linked to acquisition software. The software allowed us to study the obturation forces in real time or to store them. In this initial study, the forces developed by endodontists and students during a warm vertical compaction were analyzed. The vertical and frontware backware horizontal direction forces were first stored. Graphs of the compaction forces were then generated, permitting the analysis of the obturation method. Indeed, two cases of obturation failure were analyzed from these graphs. The mean values for the vertical forces applied by the endodontists and students were, respectively, 2.5 +/- 0.4 kg and 1.9 +/- 0.9 kg; the mean values for the lateral forces were, respectively, 0.85 +/- 0.2 kg and 1.4 +/- 0.6 kg. This device permits the analysis of compaction forces and may thus be highly useful in obtaining improvements in obturation techniques. PMID- 9220736 TI - Inflammatory response to calcium hydroxide based root canal sealers. AB - This study evaluated the inflammatory response to Sealapex, CRCS, Apexit, and Sealer 26 in the subcutaneous tissue and in peritoneal cavity of Balb/c mice. The inflammatory response of subcutaneous tissue was analyzed after 2, 4, 8, and 16 days. Intense neutrophilia was seen in response to all sealers during the initial periods. Differences among them related to the presence of necrosis and the number of inflammatory cells. In the intermediate phase marked differentiation of cells of the mononucleate phagocytic system into macrophages, epithelioid cells and multinucleate giant cells were observed with Sealapex. This response was less intense with CRCS and Apexit. Tissue necrosis was observed only at tissue sealer interfaces and only during the initial period with Sealapex but was seen throughout the experiment with all other sealers. The animals were injected in the peritoneal cavity with solutions containing the sealers and five mice from each group were killed 6 and 24 h, and 5 and 15 days later. During the initial periods (6 and 24 h) there was an intense migration of polymorphonuclear leukocytes to the peritoneal cavity in response to all sealers compared to the control. This migration was more intense for Sealer 26 and Apexit. An increase in mononucleate cell number was observed after 6 and 24 h and 5 days for all sealers and no differences were observed in relation to the control after 15 days. PMID- 9220738 TI - Thermal effects and antibacterial properties of energy levels required to sterilize stained root canals with an Nd:YAG laser. AB - Thermal effects and antibacterial properties of an Nd:YAG laser were studied to establish clinically safe levels of energy to deliver into the root canal and to determine the energy level needed to sterilize infected root canals. The results indicate that lasing cycles of 3 J-s for 15 s followed by a 15-s recovery interval can be continued for prolonged periods without risk of thermal damage to surrounding tissues. In vitro lasing of root canals inoculated with dark stained bacteria showed that two such lasing cycles sterilized only two out of eight canals, whereas when four cycles were used seven out of eight canals were sterilized. Guidelines for energy levels in endodontic Nd:YAG laser work are discussed, and base data for calculating appropriate energy levels are given. PMID- 9220739 TI - 3D reconstruction of two C-shape mandibular molars. AB - Two mandibular second molars, with an indication of C-shape morphology were processed for 3-D reconstruction. After serial cross sectioning, photographs of the sections were digitized and by using surface representation, 3D reconstruction was achieved. The first molar as the 3D reconstruction showed was single rooted with one C-shaped root canal with two foramens, while the second one was double rooted with two root canals, one C-shaped and one thin, having a common foramen. PMID- 9220740 TI - The investigation of biocompatibility and apical microleakage of tricalcium phosphate based root canal sealers. AB - The biocompatibility and apical microleakage of tricalcium phosphate based Sankin Apatite (SA) Type I, II, and III root canal sealers were investigated. Teflon tubes containing freshly mixed test materials were implanted in the subcutaneous tissue of mice. The observation periods were 24 h, 7, and 30 days, after which the areas of tissue reaction to the implanted materials were histopathologically analyzed. A dye-recovery, spectrophotometric method was used to evaluate apical microleakage. Results showed that the severity of tissue reaction among the tested materials decreased with time and at the end of the observation period both SA Type II and Type III were found more biocompatible than either Type I or Grossman's cement (GC). On the other hand, a fibrous tissue capsule was seen around the implants. There was no significant difference in spectrophotometrically measured leakage among teeth obturated with the test materials. PMID- 9220741 TI - Identification and antibiotic sensitivity of bacteria isolated from periapical lesions. AB - Periradicular tissues from 28 refractory endodontic cases requiring surgical intervention were submitted for histological diagnosis and microbiological culture. Bacteria isolated from these lesions were identified and then tested for their antibiotic sensitivity to a panel of common antibiotics. The periapical tissue specimens of 22 out of 28 lesions (79%) contained microorganisms. Of the 22 cases showing positive growth cultures, 15 were polymicrobial and 7 were single species isolates. Fifty-three different species were recovered: 29 anaerobes, 19 facultative anaerobes, and 5 aerobes. Microbes were observed under light microscopy in only one case. The most common organisms isolated were Propionibacterium acnes, Staphylococcus epidermidis, Streptococcus intermedius, Wolinella recta, Fusobacterium species, and Clostridium species. Antibiotic susceptibility results showed no clear cut evidence of significant antibiotic resistance among the species tested. The results of this study seem to corroborate earlier studies regarding the microbial population of periapical lesions refractory to nonsurgical endodontics. PMID- 9220742 TI - Defense responses of dentin/pulp complex to experimentally induced caries in rat molars: an immunohistochemical study on kinetics of pulpal Ia antigen-expressing cells and macrophages. AB - Experimental caries was induced in rats that were inoculated orally with Streptococcus mutants and maintained on a cariogenic diet. During the caries process, kinetics of the pulpal la antigen-expressing cells and macrophages was monitored immunohistochemically and was correlated with caries depth and the status of reparative dentin formation. Initial pulpal response was characterized by a localized accumulation of la antigen-expressing cells beneath the dentinal tubules communicating with the superficial caries. This was followed by a caries depth related increase of la antigen-expressing cells and macrophages in the coronal pulp. The accumulation of these cells under the dentin was most apparent when the caries had progressed into the reparative dentin. These findings suggest that the response of la antigen-expressing cells to carious irritants triggers the defense reactions of the pulp. The intensity of the defense reactions may be correlated with the permeability of carious dentin. PMID- 9220743 TI - Titanium-inlay--a new root-end filling material. AB - To obtain better results following periradicular surgery, we have developed a Titanium-Inlay with SuperEBA cement as a sealer as a new root-end filling material. This was combined with an ultrasonic root-end preparation technique. The preliminary results suggest that the surgical method we adopted is simple and the results are predictable. The material and technique are presented in this article. PMID- 9220744 TI - Some unusual clinical cases on root anatomy of permanent maxillary molars. AB - Three case reports are presented concerning permanent maxillary molars with fusion of two buccal roots. The morphology is peculiar because it is characterized by the mesiobuccal canal merging into the distobuccal canal in their apical one-third. A thorough review of the literature failed to reveal any previously reported description of permanent maxillary molars with this arrangement of canals, with the exception of a recently reported epidemiological study. PMID- 9220745 TI - Immunohistochemical detection of prostaglandins E2, F2 alpha, and 6-keto prostaglandin F1 alpha in experimentally induced periapical inflammatory lesions in rats. AB - The immunohistochemical localization of prostaglandin (PG) E2, PGF2 alpha, and 6 keto-PGF1 alpha (a stable metabolite of PGI2) was demonstrated in rat periapical inflammatory lesions induced by opening the pulp chamber. Two wk postoperatively, suppurative periapical lesions were formed, and active bone resorption was seen surrounding these lesions. Immunohistochemical examination showed that macrophages infiltrating in inflammatory tissue were positively stained for the examined PGs. In some lesions, wherein acute inflammatory changes subsided and proliferation of fibroblasts started, the fibroblasts were positively stained for 6-keto-PGF1 alpha. Osteocytes and osteoblasts were also positive for 6-keto-PGF1 alpha not only in experimental animals, but also in untreated animals. However the staining intensity of the PG in these cells was higher in periapical lesions than in normal condition. These findings suggested that the cellular sources of the PGs in the periapical lesions are mainly macrophages and fibroblasts, and that the PGs produced by these cells, and possibly osteoblast and osteocytes, may contribute to the osteolytic resorption of periapical lesions. PMID- 9220747 TI - Removal of smear layer in the root canal using oxidative potential water. AB - We investigated oxidative potential water (OPW) for its ability to remove the smear layer using a scanning electron microscope. OPW has been studied mainly in Japan and is known to suppress bacteria and viruses without harming living systems. We found that OPW used as in irrigant during and after root canal instrumentation is as effective as 5% NaOCl or 17% EDTA for opening and keeping patent the dentinal tubules. PMID- 9220746 TI - A comparison of sealer placement techniques in curved canals. AB - Sealer placement techniques have not been examined in teeth with curved canals prepared with Lightspeed instruments. Three traditional methods of placing sealer were studied, using 45 extracted human single-rooted teeth, divided into 3 groups of 15. Root canal preparations were made with Light-speed nickel-titanium, engine driven instruments. AH26 sealer was applied with either K-file, lentulo spiral, or master gutta-percha cone. Radiographs were taken after sealer placement and analyzed for amount of canal sealer fill. The teeth were then obturated with laterally condensed gutta-percha, chemically cleared, photographed, and analyzed for total canal wall sealer coverage. The results showed a statistically significant difference in canal sealer fill among the three groups before obturation, but there was no statistical difference in canal wall coverage among the three groups after obturation. None of the examined methods exceeded an average of 62.5% wall coverage of sealer after obturation. This suggests that complete wall coverage after obturation may not be possible. PMID- 9220748 TI - Measurement algorithm accuracy of the RVG-PCi in vertical and diagonal assessments at various beam energies. AB - The Trophy RadioVisioGraphy model PCi was compared to Kodak Ektaspeed Plus film for accurate recording and estimation of the length of size 15 files placed vertically and diagonally across the receptor surface. Variations in kilovoltage (50, 70, and 75) and exposure were also factored. Eight observers estimated file lengths using the proprietary software measurement algorithm for the RVG-PCi and a millimeter rule for the film-based radiographs. Both modalities resulted in slight magnification for vertically oriented files; however, the RVG-PCi caused overestimation in the order of 6 to 8% with diagonally oriented instruments. Measurement interobserver variability was least when using the RVG-PCi. It was concluded that the proprietary software supplied with RVG-PCi is not sufficiently accurate for endodontic assessment. Furthermore, exposures above 0.15 s at 75 kVp resulted in pixel saturation resulting in apparent shortening of the instrument; hence, length calculations are particularly sensitive to overexposure when using the RVG-PCi. PMID- 9220749 TI - Measurement of the cutting efficiency of endodontic instruments: a new concept. AB - The cutting efficiency of endodontic instruments was measured using an original experimental technique that incorporates new concepts to simulate clinical conditions. Five designs of #ISO 030 endodontic instruments, K-reamer (Maillefer), Flexofile (Maillefer), Helifile (Micro-mega), K-flex (Kerr), and Unifile (De Trey), were chosen and their cutting efficiency assessed at their full working length of 16 mm on two Plexiglas parallelepipeds tilted to follow the 2% conicity of the instruments. For each instrument, four series of 25 cuts were carried out and each cut made on a new flat, smooth Plexiglas surface with an even hardness of 33 VHN. Instruments were tested under a simulated clinical condition of a quarter clockwise turn ROTARY MOTION followed by a PULL ACTION at 16 mm/s rate, with a fixed load on the instrument of 325 g. Water irrigation at a rate of 85 ml/s was supplied before each procedure. Cutting efficiency was evaluated in terms of mass of Plexiglas cut (using a Mettler analytic balance with accuracy of 3 x 10(-5) g) per unit of energy used by the instrument, i.e. mg/J. Unifile was found to have the best cutting efficiency of 0.80 +/- 0.01 (Mean +/- SD) and lowest cutting efficiency loss followed by Flexofile 0.70 +/- 0.03 then Helifile 0.36 +/- 0.01 then K-flex 0.51 +/- 0.07. K-reamer was found to have the lowest cutting efficiency of 0.16 +/- 0.05. PMID- 9220750 TI - Effects of cleaning, disinfection, and sterilization procedures on the cutting efficiency of endodontic files. AB - The effects of various cleaning, chemical disinfection, and sterilization procedures on the cutting efficiency of endodontic instruments Unifile (De Trey, Bois Colombes, France), (Flexofile Maillefer, Ballaigues, Switzerland), and H File (Maillefer)) were investigated. The cross-infection control treatment procedures investigated were as follows: chemical disinfection--NaOCl (2.5%) for 12 and 48 h, and NH4 (5%) for 1 and 4 h; ultrasonic cleaning for 4 and 16 cycles of 15 min; and sterilization methods with chemiclave for 5 and 10 cycles of 20 min, Poupinel for 5 and 10 cycles of 120 min at 180 degrees C and glass beads for 10 and 40 cycles of 40 at 250 degrees C. Cutting efficiency was evaluated as the mass of Plexiglas cut per unit of energy expended by the instrument in microgram/Joule. The cutting efficiency decreased from 1 to 77%, depending on the file design and type of treatment procedures. Heat sterilization (Poupinel) did not modify the cutting efficiency of Unifile and Flexofile. The decrease in cutting efficiency was independent of frequency and duration of treatment procedures. PMID- 9220751 TI - Treatment of root fracture by CO2 and Nd:YAG lasers: an in vitro study. AB - The purpose of this in vitro study was to use scanning electron microscopy and polarized light microscopy to evaluate the feasibility of using either the CO2 laser or an Nd:YAG laser in combination with air/water surface cooling to effect fusion of fractured tooth roots. The experimental unit consisted of 81 single rooted teeth, each with an induced root fracture. Fifty-six teeth that had been reapproximated in dental stone and 25 teeth that had been reapproximated with C clamps were assigned to untreated control groups or groups for treatment using CO2 and Nd:YAG lasers. Laser treatment consisted of multiple passes along the line of fracture, which was inspected using a dissecting microscope after each pass until a visual indication of fusion or irreparable damage resulted. Scanning electron microscopy evaluation of the treated lines revealed heat-induced fissures and cracks, areas of cementum meltdown and resolidification, crater formation, and separation of cementum from underlying dentin. In no instance regardless of reapproximation technique, laser type, energy, and other parameters did the treatment effect fusion of the fractured root halves. PMID- 9220752 TI - Effects of a combination of an antibacterial agent (ofloxacin) and a collagenase inhibitor (FN-439) on the healing of rat periapical lesions. AB - To investigate the effects of a combination of an antibacterial agent (ofloxacin) and a collagenase inhibitor (FN-439) in the root canal treatment of apical periodontitis, we studied the healing process of experimentally induced periapical lesions in rats by using immunohistochemical methods. With a topical application of a combination of ofloxacin and FN-439 following experimentally induced periapical lesions, both neutrophils and macrophages became significantly decreased in number, while active cementogenesis and extensive bone formation were seen in the periapical region. However, the use of ofloxacin alone also demonstrated a beneficial effect on periapical inflammation and healing. Therefore, it is suggested that ofloxacin is powerful against bacterial infection whether FN-439 is added. The only observed effect of a combination of ofloxacin and FN-439 is that it may more effectively inhibit osteoclastic bone resorption and activate the remodeling of the apical periodontal tissue if this combined medicament is used in a stage of active bone destruction characterized by high production of tissue collagenase. PMID- 9220754 TI - Relationship between prostaglandin E2 concentrations in periapical exudates from root canals and clinical findings of periapical periodontitis. AB - In this study the relationship of prostaglandin E2 (PGE2) concentrations in periapical exudates with clinical findings of teeth with periapical periodontitis is discussed. Periapical exudate samples were obtained from root canals of 77 endodontically involved teeth during routine root canal treatment by the quantitative sampling method using paper points. PGE2 concentrations in periapical exudates (PE-PGE2) were determined by radioimmunoassay. Significantly higher levels of PGE2 were found in periapical exudates from teeth with radiolucent areas than from teeth without radiolucent areas (236.8 +/- 521.3 pg/microliter vs. 14.9 +/- 23.1 pg/microliter, respectively). The elevated PE PGE2 levels were associated with the presence of clinical symptoms that reflected an acute inflammation in the periapical lesion. In contrast, a significantly negative association of decreased PE-PGE2 levels with increasing size of radiolucent areas was demonstrated. These results suggested that PGE2 was produced locally in periapical lesions and that the PGE2 concentration in periapical exudate could reflect the state of the disease activity in periapical periodontitis. PMID- 9220753 TI - Disinfection by calcium hydroxide pastes of dentinal tubules infected with two obligate and one facultative anaerobic bacteria. AB - Bovine dentine cylinders were experimentally infected with Actinomyces israelii, Fusobacterium nucleatum, or Enterococcus faecalis. The latter is a facultative anaerobic bacteria and the others are obligate anaerobes commonly found in endodontic infections. The infected specimens were exposed to pastes of calcium hydroxide mixed with saline solution or camphorated paramonochlorophenol for periods of 1 h, 1 day, and 1 week. The viability of bacteria after these exposure times was evaluated by specimen incubation in culture medium to compare the effectiveness of the pastes in disinfecting dentinal tubules. The calcium hydroxide/camphorated paramonochlorophenol paste effectively killed bacteria in the tubules after a 1-h period of exposure, except for E. faecalis that required one day of exposure. In contrast, the calcium hydroxide/saline paste was ineffective against E. faecalis and F. nucleatum even after 1 week of exposure. The results showed that camphorated paramonochlorophenol increased the antibacterial effects of calcium hydroxide. PMID- 9220755 TI - Torsional testing of the Lightspeed nickel-titanium instrument system. AB - Revolutions to separation and maximum torque at failure of 216 Lightspeed instruments were determined in an instron using a clockwise rotation. After instruments failed, the distance the instrument separated from the tip was measured. Comparison of the results with existing ANSI/ADA specification no. 28 showed that the Lightspeed far exceeded the values of the specification for revolution to failure. On the other hand, torque to failure results showed that instrument sizes 20 and 25 exceeded the specification, whereas instrument sizes 30 through 50 were below the minimum values. Half-size instruments were not compared, because specifications for half-sizes do not exist. Comparison between mean torque values and instrument shaft diameters of the Lightspeed previously reported showed a near linear relationship up to and including instrument size 50, but overall torque to failure increased exponentially when related to shaft diameter (coefficient of determination = 0.9923). Lightspeed instruments separated 2.32 +/- 0.60 mm from the tip, generally within the land area or at the beginning of the shaft. Scanning electron microscopic observation of the Lightspeed instrument fracture site showed two distinct areas. There was a striated concentric area in the periphery of the fracture characteristic of a brittle or cleavage fracture and a corrugated area in the center of the fracture characteristic of a ductile fracture. PMID- 9220756 TI - Effects of four instrumentation techniques on curved canals: a comparison study. AB - Standardized canals (42 degrees curvature) in clear resin blocks were instrumented from size #15 to #35 by four different instrumentation techniques [SW (Senia-Wildey) instrumentation technique, "balanced force" technique, "step back" technique, and a technique combining a reaming motion and the "balanced force" technique] using Flexoreamer Batt-tip, Flex-R file, or K-Flexofile Batt tip. Changes in canal shape were investigated under standardized conditions using a computer driven testing device. Changes in shape differed significantly (p < 0.05) between the three instruments and between the four techniques (p < 0.0001) in all of the 14 measuring points. Best results were obtained when the curved canals were first enlarged with Flexoreamer Batt-tip or Flex-R file #15 and #20 using a reaming motion followed by #25 to #35 instruments using the "balanced force" technique. Overall, under the conditions of this study, this combined technique provided satisfactory canal preparation with no ledging or canal transportation. PMID- 9220757 TI - One-step apexification without calcium hydroxide. AB - Slow growth calcium hydroxide apexification may be clinically impractical in some instances. This case report describes a technique and rationale for single-step use of tricalcium phosphate as an apical plug in an immature permanent root. This method permitted immediate canal obturation and placement of the permanent coronal restoration. A 7-yr follow-up confirmed that this type of apexification could be successful. PMID- 9220758 TI - A new approach to the treatment of true-combined endodontic-periodontic lesions by the guided tissue regeneration technique. AB - Clinicians often have difficulty in the diagnosis and treatment of the combined endodontal and periodontal (endo-perio) lesion. A case of an endo-perio true combined lesion on a maxillary premolar was first treated with conventional endodontic therapy. Periodontal surgery was then completed, which included scaling and root planing and apical curettage on the tooth. The facial bony defect was then filled with a decalcified freeze-dried bone allograft mixed with tetracycline powder. A non-resorbable Teflon membrane was then used to cover the bone material and the periodontal flap sutured over this. This combined treatment resulted in minimal probing depth (2 mm), maximal clinical attachment gain (8 mm), as well as radiographic evidence of alveolar bone gain. This case report demonstrates that proper diagnosis, followed by removal of etiological factors and utilizing the guided tissue regeneration technique combined with osseous grafting, will restore health and function to a tooth with severe attachment loss caused by an endo-perio lesion. PMID- 9220759 TI - Burkitt's lymphoma mimicking an acute dentoalveolar abscess. AB - Burkitt's lymphoma is a monoclonal proliferation of B lymphocytes classified histologically as a poorly differentiated lymphocytic lymphoma. The jaw and retroperitoneal structures are the most commonly involved sites. Prognosis is highly dependent on the stage of the disease. In some cases, the first manifestation of Burkitt's lymphoma is in the jaws, and symptoms may be misdiagnosed as infection. Dental radiographs can play an important role in the diagnosis. A case of a peculiar Burkitt's lymphoma involving the mandible that was misdiagnosed as an acute dentoalveolar abscess is presented. PMID- 9220760 TI - Treatment of stripping perforations. AB - Strippings are problems that are frequent on thin and concave roots. Treatment and prognosis differ from that of a lateral root perforation because of the size, oval shape, and thin edges of the striping. We propose a two-step technique: an endodontic phase in which the root canal system is sealed with gutta-percha overflowing through the stripping perforation and a surgical second step that will allow elimination of this excess. PMID- 9220761 TI - The use of tungsten carbide needle holders to remove intracanal objects. AB - The removal of metallic filling materials and posts is necessary before endodontic retreatment. Retreatment in such cases can require considerable time and effort. This article presents a technique to facilitate the removal of metallic obstructions from root canals using tungsten carbide needle holders. Clinical cases demonstrate the effectiveness of this technique. PMID- 9220762 TI - Managers need to listen. PMID- 9220763 TI - Don't ask, don't know. PMID- 9220764 TI - Classroom and clinical teaching in nursing: delineating differences. AB - Although ability to teach in the classroom is generally considered sufficient preparation for clinical teaching, the reality can be a surprisingly different multidimensional role. The author presents a list of 15 specific and significant differences that exist between the two teaching performances. The settings are compared using three components common to both: instructional, evaluative, and interpersonal interactions. Myths, misconceptions, and realities about the two teaching arenas are discussed. A conceptual relationship between the two instructional settings is presented. Recommendations are offered to reduce teacher anxiety, confusion, and frustration created by existing myths/misconceptions. PMID- 9220765 TI - Channeling nurses' anger into positive interventions. AB - Nurses are angry, and with good reason, but few seem to be proficient in anger management. Work-related experiences of anger were vividly described by female registered nurses (Smith, Droppleman, & Thomas, 1996) and male registered nurses (Brooks, Thomas, & Droppleman, 1996) in phenomenological interviews. The roots of the anger were found in the system and the interpersonal and intrapsychic elements of nursing. In this article the authors outline strategies to address system and interpersonal sources of anger provocations, and describe positive interventions that can reduce the intrapsychic characteristics that fuel personal anger. PMID- 9220766 TI - Providing primary care to poor urban women. AB - Public health nurses used the findings of a 1989 community survey to develop a program to meet the healthcare needs of an urban population in western Pennsylvania. The article details the expansion of the services beyond the storefront location to provide comprehensive women's care through the Rainbow Bridge Project. A formula for other public health nurses to use in replicating a neighborhood center, pointing to the need for many skills not traditionally considered nursing, such as networking, fund-raising, and public relations is illustrated. PMID- 9220767 TI - Ambiguity in the "Nursing Statement on assisted Suicide". AB - Nursing is multidimensional, interactive, interdisciplinary, and complex. Almost anything that can be said about nursing can be said another way. Some things worth being said and heard will not follow the norms of journal presentation. A forum accommodates the emerging voice, the new format, the innovative approach. Nursing Forum, in an effort to honor the "independent voice" in nursing, presents here the voice who elects to enter the dialogue, but who does so "in another way." PMID- 9220769 TI - Mental health nursing education within Project 2000 does not work. PMID- 9220768 TI - Is there room for idealism in nursing today? PMID- 9220770 TI - Age of lost innocence. PMID- 9220771 TI - Caution: baby on board. PMID- 9220772 TI - Stage set for equality claims. PMID- 9220774 TI - A job title. PMID- 9220773 TI - Emergency measures. PMID- 9220775 TI - Spot the carer. PMID- 9220776 TI - The young ones. PMID- 9220777 TI - Guilt complex. PMID- 9220778 TI - Informed care. PMID- 9220780 TI - Shock absorbers. PMID- 9220779 TI - In from the cold. PMID- 9220782 TI - Good old days. PMID- 9220781 TI - The morale of the story. PMID- 9220783 TI - The National Health Service is not in very good shape. PMID- 9220784 TI - Taking research and development forward. AB - The funding of research and development in NHS trusts has changed since the Culyer report. This has increased opportunities for nurses to become involved in R&D at different levels. This article explains the distribution of money and makes recommendations for partnerships in R&D. PMID- 9220785 TI - Close observation: how to improve assessments. AB - Close observation on an acute psychiatric ward is an intervention of restraint. It is intended to ensure the safety and well-being of the patient. This article presents research into the prescription of dose observation and sets out to offer a model for prescribing and reviewing this intervention. PMID- 9220786 TI - The dangers of cutting school nursing services. AB - This article highlights the important role played by the school nursing service. it stresses the need for school nurses to collect and analyse data so that they are able to provide evidence of their effectiveness. It also warns against the long-term implications for the nation as a whole, and its youth in particular, if nursing cuts are made in the interests only of short-term expediency. PMID- 9220787 TI - Losing sight of the future. PMID- 9220788 TI - Formula baby milk: what are the facts? PMID- 9220789 TI - Hull of a good job. PMID- 9220790 TI - Fractured neck of femur. The role of the nurse. PMID- 9220791 TI - Opportunity knocks in the community. PMID- 9220792 TI - Career focus on Kate Billingham. PMID- 9220793 TI - [The life activity. Sleep is an important factor in recovery and staying well]. PMID- 9220795 TI - [Sleep...]. PMID- 9220794 TI - [Conditions for the patients' sleep from the nurses point of view]. PMID- 9220797 TI - Need mor mo' on the Child Health Insurance bill. PMID- 9220796 TI - [Nutrition and feeding of the dying with special reference to dehydration in the last phase of life]. PMID- 9220798 TI - Can NCAST and HOME Assessment Scales be used with Hmong refugees? AB - This descriptive study of a convenience sample of 32 recently settled, preliterate Hmong families tested the applicability of the NCAST and HOME assessment tools in the Hmong refugee population. The sample means on the Nursing Child Assessment Feeding Scale (NCAFS), Nursing Child Assessment Teaching Scale (NCATS), and the Home Observation for Measurement of the Environment (HOME) were compared to a randomly selected sample of white (n = 60), African American (n = 60), and Hispanic (n = 60) families from the University of Washington NCAST (Nursing Child Assessment Satellite Training) normative data bank. The NCAFS and NCATS Hmong total means were not significantly different when compared to the combined normative sample total means. However, when the ethnic groups were compared using ANCOVA with ethnicity and education as covariates, the parent, child, and scale total means were found to be significantly different (p < .05). Hmong HOME total means were significantly lower when compared to the combined normative sample (p < .05) and with separate ethnic group comparisons (p < .05). Although lower for this population lacking formal education, scores were not significantly different when compared to the combined normative sample means. Findings suggest that the NCAST tools should be used with respect for the impact of ethnicity and education on total scores. Findings do not support the use of the HOME scale with this population except as a teaching guide. PMID- 9220799 TI - Hmong healing practices used for common childhood illnesses. AB - Central California has become a place of refuge and settlement for Southeast Asian Hmong immigrants over the past 20 years. The resulting assimilation of this new cultural group into American society has produced the need for multi-ethnic understanding and culturally sensitive health care interventions. Knowledge regarding the health care practices of Hmong parents provides the foundation for this understanding and an integrated and meaningful model of child care. This study of Hmong parents describes (a) their indigenous healing practices, (b) the purposes of these health care practices, (c) their beliefs about western medical care, and (d) their views on seeking western pediatric health care. A one-group descriptive design was used. Interview data from Hmong parents (N = 21) describes illness causation, healing rituals, herbal remedies, and other health care traditions. PMID- 9220800 TI - Maternal psychosocial factors associated with substance use in Mexican-origin and African American low-income pregnant women. AB - PURPOSE: To describe ethnic-specific patterns of substance use before and during pregnancy in low-income pregnant women, examine the associations between psychosocial factors and patterns of substance use within ethnic groups, and assess maternal sociodemographic, prenatal, and psychosocial factors of women who continue to use substances during pregnancy and those who do not. METHOD: A prospective study of low-income, primiparous African American (n = 255), Mexican American (n = 525), and Mexican immigrant (n = 764) women was conducted in 22 prenatal care clinics in Los Angeles, CA. Data were collected in face-to-face interviews in both English and Spanish on prenatal life events, anxiety, sources of support, and substance use behaviors three months before and during pregnancy. FINDINGS: Significant ethnic differences were found in use of alcohol, cigarettes, and illicit drugs. African American women were more likely than Mexican-origin women to report use of substances before and during pregnancy. Mexican American women were more likely than Mexican immigrant women to report use of substances before and during pregnancy. Women who continued to use substances during pregnancy were less likely to be living with the baby's father, to have planned the pregnancy, to report having been able to go for prenatal care as soon as they wanted, and more likely to be identified at medical risk. CONCLUSIONS: Providers must increase the assessment and monitoring of substance use behaviors of low-income women in prenatal care settings. The role of health care providers must encompass advocacy and public health education. PMID- 9220801 TI - Development of a research-based standard for assessment, intervention, and evaluation of pain after neonatal and pediatric cardiac surgery. AB - This analysis of retrospective and prospective data quantified children (age range 0-18 years, total n = 132) during their stay in a cardiothoracic intensive care unit and examined pain management and sedation practices. Data on both factors that could potentially affect pain and its management, and analgesics/sedatives ordered for and administered to subjects were collected from chart review. In the prospective group, pain intensity was measured twice daily using the Wong-Baker FACES Pain Rating Scale. Repeat cardiac surgical procedure subjects reported significantly more pain than nonrepeat subjects on the first postoperative night. Subjects with sternal incisions reported significantly more pain than subjects with submammary incisions. Not all subjects were premedicated with analgesia for invasive procedures. Significantly greater amounts of analgesia were received by the 0-3 year-old subjects. Large amounts of sedation were used, especially in children under 3 years of age. The results prompted development of a nursing standard to assess and manage pain and sedation in this population. PMID- 9220802 TI - Home parenteral nutrition (HPN) in children in Sweden. AB - This study represents a national survey of all children identified on home parenteral nutrition (HPN) in Sweden via all hospital pharmacy distributors. Indications for HPN from this sample were chronic intestinal pseudo obstruction (n = 5) and short bowel syndrome (n = 7). Linear growth was normal in 9 children and all school-age children attend classes at age-relevant levels. Families appear to have adapted well. There was one divorce among these families, and over the course of the HPN child's illness, younger siblings were born. PMID- 9220803 TI - ABCs of health insurance benefits: a practical guide. AB - Understanding health insurance benefits is essential for coordinating the continuum of care for families. All plans have basic areas of coverage but vary in the amount of service provided. Educating families on health insurance benefits is the key to empowering families as consumers. PMID- 9220804 TI - Johnny's story: transfusing a Jehovah's Witness. AB - Jehovah's Witnesses refuse blood transfusions for themselves and for their children. This action can be difficult for health professionals to understand and can lead to tensions between the staff and family. For one family, their refusal of blood for their child lead to a greater understanding of their religion and its beliefs for those who cared for them. Interspersed with their story are the medical reasons their son required blood, the reasons Jehovah's Witnesses refuse blood transfusions, and what the acceptable alternatives are to Jehovah's Witnesses. This article will share the thoughts and feelings of the family and the nursing staff who cared for the family during this crisis. PMID- 9220805 TI - Pediatric management problems. Acute asthma attack. PMID- 9220806 TI - The FLACC: a behavioral scale for scoring postoperative pain in young children. AB - PURPOSE: To evaluate the reliability and validity of the FLACC Pain Assessment Tool which incorporates five categories of pain behaviors: facial expression; leg movement; activity; cry; and consolability. METHOD: Eighty-nine children aged 2 months to 7 years, (3.0 +/- 2.0 yrs.) who had undergone a variety of surgical procedures, were observed in the Post Anesthesia Care Unit (PACU). The study consisted of: 1) measuring interrater reliability; 2) testing validity by measuring changes in FLACC scores in response to administration of analgesics; and 3) comparing FLACC scores to other pain ratings. FINDINGS: The FLACC tool was found to have high interrater reliability. Preliminary evidence of validity was provided by the significant decrease in FLACC scores related to administration of analgesics. Validity was also supported by the correlation with scores assigned by the Objective Pain Scale (OPS) and nurses' global ratings of pain. CONCLUSIONS: The FLACC provides a simple framework for quantifying pain behaviors in children who may not be able to verbalize the presence or severity of pain. Our preliminary data indicates the FLACC pain assessment tool is valid and reliable. PMID- 9220807 TI - Sibling rivalry and the new baby: anticipatory guidance and management strategies. AB - Sibling rivalry can be found in many families and frequently creates a stressful and challenging situation for parents. The arrival of a new baby often causes older siblings to feel displaced, frustrated, angry, and even unloved. Age, gender, personality and temperament, and parental behavior are factors that appear to influence the degree to which sibling rivalry occurs. Common reactions of older siblings to the birth of a new baby include aggression toward the newborn, behavioral regression, and attention seeking behavior, as well as independence and maturity. Anticipatory guidance is recommended to help parents adequately prepare their older child for the arrival of a new sibling. Strategies for managing sibling rivalry include open parent-child communication, equal treatment of siblings, non-intervention in sibling conflicts, distraction, and separation. Parents can minimize feelings of jealousy between siblings by providing a supportive, nurturing environment that allows each child to feel secure and loved. PMID- 9220808 TI - Parent to parent support and health care. AB - Parent to Parent programs offer a unique model for personalizing family support services according to family preferences. Over 500 of these programs across the United States provide emotional and informational support through careful matching of veteran parents with referred parents who have a child with a disability. A national survey of Parent to Parent program coordinators and participating parents examined referral sources, program supports, program demographics/descriptions, and examples of best practices. Parent to Parent programs can complement health care services by empowering parents through information and support and should be an essential component of a comprehensive family support system. PMID- 9220809 TI - Pediatric asthma update: current therapy and new agents. PMID- 9220810 TI - Children's health legislation introduced, linked to tobacco tax. PMID- 9220811 TI - New report of State of America's Children finds child well-being lagging despite economic recovery. PMID- 9220812 TI - [Families are Germany's largest and least expensive nursing service]. PMID- 9220813 TI - [Thousands of jobs in geriatric care are endangered]. PMID- 9220814 TI - [A close look at latex gloves. 2. The union demands protective measures for nurses]. PMID- 9220815 TI - [Quality assurance on the ward. The use of nursing standards improves the quality of care]. PMID- 9220816 TI - [Care and spiritual assistance together help the families]. PMID- 9220817 TI - [Hyperthermia. A new therapeutic possibility in oncology makes special demands on nurses]. PMID- 9220818 TI - [The fallacy about planned care]. PMID- 9220819 TI - [The social environment has an effect on health and disease. Nurses should be aware of all of the patient's environment]. PMID- 9220820 TI - [The nursing process guidelines at the Frankfurt University Hospital. The guidelines do not contain measurable criteria for quality assurance]. PMID- 9220821 TI - [Nursing guidelines]. PMID- 9220822 TI - [Strategic training of personnel in health care. 1. Friendliness to clients will determine the hospital's stature]. PMID- 9220824 TI - [Introduction of health care planning on the basis of the nursing models according to N. Roper, W. W. Logan, A. J. Tierney. A project of the Pediatric Department of the University of Hamburg]. PMID- 9220823 TI - [Holistic maternity and child care: a difficult path towards appropriate and timely care for mother and child]. PMID- 9220826 TI - Antenatal counselling for the haemoglobinopathies. PMID- 9220825 TI - Why don't British women breast feed for longer? PMID- 9220827 TI - Grief and the concept of loss in midwifery practice. Part 1. Normal and abnormal grief. PMID- 9220828 TI - Summer safety. PMID- 9220829 TI - Vital signs: spotting liver disease in babies and children. PMID- 9220830 TI - Selecting inhaler devices for children with asthma. PMID- 9220831 TI - Oral care in the early years. PMID- 9220832 TI - Handling hayfever. PMID- 9220833 TI - Tackling a visitation of threadworms. PMID- 9220834 TI - [Epidemiology of violence--an approach to the problem of violent death in contemporary Brazilian society--the case in Santa Catarina]. AB - This paper discusses the problem of violence and its expression upon mortality due to external causes. A few indicators are offered, which have been worked upon it to emphasise the importance of the theme. In a general way, the study demonstrates violent death has had its magnitude increased along the years, not only throughout Latin America but also in Brazil and in Santa Catarina. PMID- 9220835 TI - [Sexual orientation guide for patients with spinal cord injuries. Applied methodology and results]. AB - The present work refers to the elaboration of a sexual orientation guide to spinal cord injured patients done by a nurse from the rehabilitation team in the Rehabilitation Medicine division at Sao Paulo University Medicine School Clinics Hospital. Its purpose is clarifying issues concerning sexual adjustment and helping those patients to express their sexuality in an appropriate way. Besides, nursing team can use it to improve rehabilitation nurse therapy even during health care educational groups. PMID- 9220836 TI - [New technologies and techniques in the care of stomas]. AB - The effective nurse role, stomatherapist or not, to select the devices used by the ostomy patient is only possible with the support of advanced technological improvements reached by the specific collecting systems in the stoma care and which are commercially available. With technological advances reached and associated with the proportional technical evolution, it is possible to give greater care to the stomas which will be ultimately reflected in the ostomy patient quality of life. Considering the technique in stoma care, we emphasise that the nurse, stomatotherapist or not, must be familiar with the collecting systems commercially available in order to make an adequate selection for the ostomy patient. Technological advances as well as technical evolution in the stoma care are responsible for the harmony of the triad ostomy/peristomal skin/collecting system used, facilitating self-care, improving quality of life and giving support not only to the physical rehabilitation but also to the ostomy patient psychological and social life. PMID- 9220837 TI - [Autologous blood transfusion using preoperative collection and intraoperative recovery in orthopedic elective surgery]. AB - Between the period of January 1994 and June 1995, the results of Autotransfusion Program of the Hospital do Aparelho Motor SARAH-BSB were evaluated. The goals of the program are: 1. Minimise and, if possible, eliminate the risks of homologous transfusion; 2. Increase the security, efficiency and cost to benefits relation of the hemotherapic procedures; 3. Minimise the demand of blood products for those patients with difficult compatible blood to be found. The program has received 194 patients, among these, 100 have been selected for this study because they had had blood collection and surgery concluded. The average age of the patients was 34.3 years (11/72 years) median was 30 years and mode 15 years. The patients have undergone elective orthopaedic operations. Autologous transfusions have been made by predeposit and intraoperative salvage (Cell Saver). Six patients (6%) presented mild reactions associated to blood drawn. Additional homologous blood was needed to 20 (20%) patients. Our experience demonstrates the security and efficiency of transfusion in patients from childhood until elderly. It recommends the creation and expansion of existing programs using autologous transfusion in services with elective surgery. We also consider that nursing care stimulates patients participation in the program. PMID- 9220838 TI - [The use of papain in plantar ulcers]. AB - This work has as a goal to contribute to decrease the inability in leprosy and continuous recurrence of plantar ulcers, through the use of a treatment method using papaine and actions of health education. This work has been done in a health centre with patients that presented plantar ulcers and agreed to participate in the proposed treatment. Analysing and comparing the obtained data before and after treatment, a greater adhesion of patients to this treatment, a quicker healing in relation to other methods used before and a greater interaction with the patient has been observed. PMID- 9220839 TI - [Treatment of venous stasis ulcers with Unna's boot and activated charcoal]. AB - In this study the author describes the results of venous stasis ulcers treatment using Unna's boot and activated coal and silver in 47 patients aged 30-80. Total treatment time was from 2 to 16 weeks, with the average of 6 days between two consecutive bandages. Among these 47 patients, 87% have had their wounds healed, 8.5% have been unsuccessful in healing them and 4.5% have dropped the treatment, the main profits observed were the decrease of bandage number, patients and Public Health Service money saving and treatment compliance. PMID- 9220840 TI - [Determination of air volume in the entotracheals tube cuff]. AB - The aim of this study is to determine the best regulation of air volume to be inflated into cuffs of different endotracheal canula diameters of high residual volume and low pressure. Maintaining the pressure exercised by a balloon in the trachea mucous membrane between preconized limits of 20 to 30 mmHg (less than the trachea capillary pressure). Therefore, 10 ml of air have been inflated into the canular cuff of intubed patients with different internal diameters (7.0-7.5- 8.0 8.5-9.0) one by one, measuring each millilitre of inflated air, the corresponding to pressure of the cuff in the trachea mucous membrane. The monitoring of the pressure has been made by a previously calibrated manometer. The procedure has been performed in 25 adult patients of both sexes, grouped in numbers of 5 for each internal tube diameter. The results have been analysed through linear regression analysis which has shown a close relationship between ml of inflated air and pressure in the cuff by tube diameter. After analysing the results, the author indicated, by means of graphic representation, the quantity of air to be inflated in the cuff and respective exercised pressure on the trachea mucous membrane, by tube meter, in a way to offer a contribution to the staff who provides assistance to intubed patients, in the sense of minimising the possible injuries and complications and assure a better respiratory ventilation. PMID- 9220841 TI - [The meaning of parenteral nutrition for the hospitalized patient]. AB - The need to understand the meaning of parenteral nutrition to the client in hospital context has come from my personal questioning on nursing care. Participative observations have been made as well as interviews and diligences with seven clients in hospital, who have been having parental nutrition. These information have been analysed through interactive approach, showing the situation faced by patients who have hospital assistance; how they say parental nutrition within this context; and what meaning has been given to it. This study revealed feelings, behaviours and attitudes related to the biological, emotional and social cultural aspects of the patient. All of them have been linked to parental nutrition. The limits faced by parental nutrition and its obstacles in the nursing assistance were finally understood. PMID- 9220842 TI - [Perceptions of unskilled patients of self-care bed baths]. AB - Descriptive study with qualitative approach performed on the medical surgical and orthopaedic treatment units with the objective of verifying and discussing the acceptance/rejection/satisfaction of bath in bed from the patients themselves perception. The population is composed of self-care non-skilled patients. The semi-structured interview and direct observation have been used for data collecting. Through reports, we have been able to verify that patients perception on bath in bed were diversified, considered disagreeable and constraining by some and indispensable by others. The aspects on the water quality, soap, nurses slowness and more humanized relationship have also been described. PMID- 9220843 TI - [Nurses dealing with emergency patients: a psychodynamic approach]. AB - The purpose of this research is to investigate the existence of defensive strategies employed by intensive care unit nurses at a public hospital in Brasilia-DF, manage the suffering generated in the nurse-patient relationship, which arises a constant confrontation with life-death contents. This effort involved four collective interviews with eight professionals. The interviews theme were the difficulties, limitations and fulfillment with the patient care. The defensive behavior were analysed through the professionals' speeches. Four strategies were identified: impersonalness, emotional distance, avoidance of communication and high evaluation of technical procedures. The employment of these strategies facilitates a psychological balance to minimize the contact with suffering, although, these defensive strategies can affect quality of work or the nurse's life extra work. PMID- 9220844 TI - [Workshop: "Woman--a trip through her body"]. AB - Morbidity due to cervical and breast cancer in Brazilian woman is high, despite the fact that these pathologies are preventable when detected early. The coverage of public health in this disease is unsatisfactory specially if one takes into account the fact that the techniques used for early detection and prevention of those types of cancer are simple and inexpensive. We observed that not many activities have been developed aiming at the education of women with regard to self-knowledge of their bodies. As a result, we prepared in 1993 a workshop entitled "Woman, a trip through her body" comprising 6 group dynamics where there is an intense participation of every woman as it is a very exciting process. The workshop is carried on by one member of the faculty staff, three nurses and nursing students of Federal University of Espirito Santo. There are 30 participants per workshop each one receiving a name tag. This workshop is composed of the following group dynamics which are developed in a cosy and informal atmosphere: Dynamic 1-Pocket History; Dynamic 2-Hands Touch; Dynamic 3 Drawing/ Modelling; Dynamic 4-Trip through the Body; Dynamic 5-Self-examination; Dynamic 6-Body Feeling. PMID- 9220845 TI - [Analysis of the level of knowledge in the population of the teratogenic effect of alcohol and the activities of nurses]. AB - Alcohol which, at first, is part of great ceremonies, parties, get-togethers, is now becoming an important issue, for the number of women who consume alcoholic beverages has increased, and consequently the number of pregnant women, considering that not only the mother's body but also the one of the child that is being formed, are under metabolic changes, and inevitably will be a target for acquired alcohol alterations (intake). During the human development, it is important to take into consideration the pregnancy stage and depending on it, the individual becomes susceptible to teratogenic agents. This paper aims at providing some contribution related to this social issue bringing knowledge on the action of alcohol on the fetus alterations, from the slightest (irritability, lack of attention, mobility deficiency) until Fetal Alcoholic Syndrome (F.A.S.) and also tries to analyse the extent of people knowledge on the consequences of alcohol effects over pregnant women bodies and the possible changes that may occur with the fetus. In order to make this study possible, interviews have been made by the use of forms with a population sample of about 100 people living in the northern, western, southern and interior low land regions. We have also visited some public and private institutions (23) on those regions, interviewing nurses in paediatric wards, aiming at identifying the difficulties found related to the diagnosis and management towards F.A.S. PMID- 9220847 TI - [The disinfection of nebulizers in a basic health unit in Ribeirao Preto]. AB - Considering the widely divergent views of the nursing staff with respect to nebulizer disinfection in Basic Health Units (BHU) and to the lack of standardization of the use of chemical products and of the disinfection process as a whole, the author undertook an investigation with the following objectives: to observe systematically the routine procedure of nebulizer disinfection in the aerosol room of a BHU, and to detect, point out and warn about possible faults in the disinfection process. The disinfection process was observed directly and systematically and a questionnaire with open questions was applied to the nurse responsible for the service. The results showed that the process of disinfection performed is not consistent with recommendations by the Health Department and/or State Health Secretariat, and that the personnel involved in this activity has incomplete knowledge of the standardization of the use of the chemical products utilized and of the conditions that interfere with their action during the disinfection process. The results obtained indicate the need for a standardization of the disinfection process as whole, according to current sanitary norms and for continual in-service education in order to improve the quality of nursing care provided to the users of health services. PMID- 9220846 TI - [Ways of prevention and control of urinary hospital infections in the hospitals of the city of Saint Paul]. AB - Theorectical knowledge and practical ability of nurses and nursing auxiliaries regarding methods of prevention and control of hospital acquired urinary tract acquired infection are studied in 29 hospitals of Sao Paulo. For this it was filled a form with questions constructed with questions for the correct form of prevention and control of urinary tract acquired infection, particularly those that were published for the CDC guidelines of the prevention and control of the Center of Disease Control (CDC). We also studied the possible effects of the infection control Department on the Theoretical Knowledge and practical ability of nurses and nursing auxiliaries regarding infection control measures. PMID- 9220848 TI - [Nursing in a pediatric unit. Proposal for systematization]. AB - The present work reports an initial process of nursing assistance systematization developed in a Pediatric Unit of a medium size hospital in Sao Carlos-SP, considering the actual human resources and the type of approach adopted by the institution on the assistance given to the hospitalized child. A Routine Manual was elaborated based on the literature and with the staff participation, and after that, a training was realized with these employees, starting from routines previously sketched, using groupal dynamics that looked forward to rescue the knowledge that they already had about the themes. The work considered the employees opinion about their participation in the systematization process. PMID- 9220849 TI - [Evaluation of a course for nurses aides by the students]. AB - Treats from comparative study who seeked to investigate how the students from two groups evaluate the courses of auxiliary in nursing by they coursed all that quality, duration, training and contents of the theoretical class. The seeks of the datas the result went realized through from the questions distribute finish of courses. The results demonstrated who the biggest peace the two groups judged the quality of course excellent, the duration, the training and contents of the theoretical class sufficient and aimed the library who a recourse very utilized for somebody in to pass of the courses. PMID- 9220850 TI - [Oral communication with hematologic patients: facilitating and blocking behaviors]. AB - In this descriptive study we adopted the model of FORREST (1983) for the categorization of oral utterances by the members of the nursing team when caring for hematological patients with oncological alterations. In order to identify the frequency of facilitating(F) and blocking(B) categories of communication and of their subcategories, we videotaped and observed the interactions between 8 patients and 14 members of the Nursing team in a Teaching Hospital in the city of Ribeirao Preto-SP. We recorded 8822 categories, 56.9% of which were facilitating. The most frequent of these were: providing information (50.3%), clarifying (28.0%) and recognizing the presence (11.2%). The most frequent blocking categories were: closed questions (64.6%), giving advice (13.9%) and approving or agreeing (10%). The remaining F and B subcategories were observed at frequencies of less than 4.5%. We suggest the more frequent use of other facilitating categories and the use of closed questions in special situations. We believe that, for a more profound relationship between nursing team and patients, it is of fundamental importance to reduce the "giving advice" and "approval" categories as they are being used now. PMID- 9220851 TI - [Opinion of alumni about the teaching of nursing diagnosis according to North American Nursing Diagnosis Association]. AB - In accordance to North American Nursing Diagnosis Association (NANDA), nursing diagnosis is a clinical judgement of the individual, family or community answers to the vital process or to the actual or potential health problems. These gives the basis for the intervention selection by which the nurse is responsible. Believing in the importance of nursing diagnosis, its teaching was introduced 4 years ago in the subjects of Woman s Health Assistance and Perinatal Nursing, in the 6o. period of the undergraduate course in nursing at UNICAMP. Aiming at evaluating these experience, we proposed to get the students opinion and obtain subsides to perfect teaching. Teoric teaching of 4 hours was given and it was indicated the basic bibliography. The supervised practical application was developed during 21 hours in a neonatal unit and 45 hours in health centers. At the end of the subjects, 16 students answered a form with closed and open questions. All the students considered valid the study of the nursing diagnoses. Most of the students considered more proper to introduce the teaching in the 3o. semester, when the nursing basic procedures is begun. As to the difficult of learning in the use of diagnoses, 68.7% of the students reported a difficult level higher to 25% and lower to 75%. The students group considers that the study of nursing diagnoses brought modifications in its concept of nursing assistance. PMID- 9220852 TI - [Intuition: a discussion in the literature]. AB - The purpose of this article is to present the concept of intuition in the literature in general and review how this theme has been considered in the nursing literature in particular. The findings in the literature where intuition appears as an innate and accessible attribute of the human being are presented in short. The authors assume a favorable position concerning the association of the rational and intuitive aspects of the human being as a kind of personal development and amplification of the possibilities to understand and help people. PMID- 9220853 TI - [Paradigms in obstetric nursing]. AB - The course of childbirth care practice in England and in the United States of America can be described by focusing on the relationship between the incipient nursing profession and the traditional profession of midwife, throughout the XVIIIth and the XIXth centuries. This paper proposes the study of such a relationship by adopting the Greek mythology goddesses as archetypical figures of female behavior. It relates the nurse to the goddess Athena, protector of the arts, the cities, the patriarchal values, the status quo-the personification of the father's daughter archetype-and the traditional midwife to Artemis, goddess of the hunt and the moon, protector of the wilderness, the weak, and the young the personification of the great sister archetype. Under such a perspective, it deals with the decline of the traditional midwife practice in those countries. Finally, it poses the question of the obstetrics nursing pattern as something to be constituted in conformity and in complicity with the women's organized movement and their claims in the field of health. PMID- 9220854 TI - [Socioeconomic and educational characterization of nursing students in the schools of Minas Gerais]. AB - This study compare the profile of nursing students from public and private schools located in the State of Minas Gerais, Brazil. It shows the similarities and differences between the two groups. The results show that the most significant differences are related to school life, requisites for learning and the economic situation of the students. The private schools student is in disadvantage, he enters at university later, disposes less time to study, the majority works, his parents have less regular instruction. This results can be used by other professionals to continue this kind of research with a broader scope. PMID- 9220855 TI - [The effect of massage before venipuncture on the reaction of pre-school and school children]. AB - The objective of this work is to describe the effect of massage done by parents on the reaction to venous puncture of preschooler and school age child hospitalized. Children's reactions were evaluated through the data of vital parameters, non-verbal communication and verbalization. The results obtained indicated that massage had significant effect in non-verbal reactions, especially those related to muscular relaxation. Effects on the reaction of vital parameters showed no difference between the two procedures, with and without massage, realized on the same child. PMID- 9220856 TI - [The phenomenon of recruiting and selecting nurses in a hospital: a phenomenological focus]. AB - This study had the purpose of showing the recruitment and selective process of nurses from their own experiences going through this process. The phenomenology was used as the methodological course, the phenomenon was based on the references by Joel Martins. The study was done from the speeches of eight nurses who went through the recruitment and selective process. The analysis of the speeches followed the moments of the phenomenological description, reduction and understanding until revealing the essence of the recruitment and selective process of nurses. From the phenomenological analysis of the speeches it was possible to get the aspects that represent the general understanding of the recruitment and selective process of nurses. In this way, the essence of the phenomenon was characterized by the relation among the man, the Company and the society, these relations were influenced by several determinants beyond the situation. Through this perspective, new horizons were opened concerning the thoughts about the recruitment and selective process of the nursing staff. PMID- 9220857 TI - [The quality of life of diabetics]. AB - The purposes of this study were to identify the meaning of the quality of life for the people who suffer from Diabetes Mellitus, to recognize the aspects which affect most their lives due to this disease and the degree of the satisfaction in their lives as well. Participated in this research forty-six (46) diabetic patients, adults of both sexes, who were in a polyclinic for treatment. The results showed that the meaning of the quality of life had priority related to the physical well-being (54.5%), to the social and economical stability (26.0%), and to the spiritual and emotional well-being (16.9%). The most affected aspects due to this disease were: studying and home activities (38.5%) physical ability (25.6%) and family relationship (10.3%). Concerning about the degree of satisfaction with their lives, the majority of them (66.6%) considered themselves satisfied or very satisfied. It is worth while to point out the importance of considering the multimensionality of the concept of quality of life while attending the diabetic person. PMID- 9220858 TI - Classification of nursing practices in collective health in Brazil. PMID- 9220859 TI - The climacterium and its meaning for women. PMID- 9220860 TI - [Tobacco consumption and nicotine dependence. A study of adolescents]. AB - A look at the prevalence of tobacco use and the degree of nicotine dependency in a group of 14 to 18 year olds. The authors emphasize that this age group needs to be presented with preventative smoking activities in order to forestall tobacco addiction; it has been clearly demonstrated that it is much easier to prevent tobacco dependency than to eliminate it. PMID- 9220861 TI - [Vitamin deficiencies. Repercussions on the health of the female population]. PMID- 9220862 TI - [Evaluating the risks of pressure ulcers. The Braden scale]. AB - An article which revises the scales used when evaluating the risks pressure sores, the context in which this might arise, and its value in our country as a useful criteria when determining which scale to choose. Also presented is a translation of the Braden Scale into Spanish as well as a comparison of its difference with respect to the Norton Scale. PMID- 9220863 TI - [A centenary: the first nursing school "Santa Isabella de Hungria"]. PMID- 9220865 TI - [Cervical vertebrae]. PMID- 9220864 TI - [Diabetes mellitus. Expectations in future health education]. PMID- 9220866 TI - [Fracture of the proximal third of the femur. 1. Preoperative measures]. AB - Fractures of the proximal third of the femur occur in great numbers, especially among the over 65 population. Due to the serious nature of these injuries it is imperative that nurses have specific procedures and care plans to deal with them. The authors approach this topic in a two part series: the first part deals with the preoperative period, and the second part looks at postoperative care. PMID- 9220868 TI - [Clinical monitoring. A method for quality control]. AB - Clinical monitoring approximates the quality control standards that have been developed by the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) and utilizes clinical indicators to measure the relative quality of the health care given. This method can be used to compare the quality of desired care against actual day by day care. To demonstrate how it can be applied, an example of clinical monitoring performed in a U.S. hospital is presented. PMID- 9220867 TI - [Bacteriuria during bladder rehabilitation in patients with spinal cord injuries]. PMID- 9220870 TI - [A-Z: talking about dictionaries]. PMID- 9220869 TI - [Drug administration by inhalation systems]. AB - Inhalant systems are often the drug administration method of choice for numerous respiratory diseases. The use of these systems is increasing due to their many advantages, one of which is the small quantity of medication they can deliver to a specific area. However, they also have their disadvantages, as inhalers require certain techniques that are not always followed. Inhalants may be propelled by a variety of methods: pressurized canisters, pumps, dry powder dispensers, nebulizers. Because each of these systems have their own administrating technique, the patient needs to be instructed in the proper use of the inhaler, with periodic follow-ups by health care workers to insure this. PMID- 9220871 TI - [The practice of writing]. PMID- 9220873 TI - [Hospital refuse. Safety in the laboratories]. PMID- 9220872 TI - [Verification of the agreement between the personnel of a first aid group concerning triage]. PMID- 9220874 TI - [Postoperative pain control in the Trento community hospital. Perception of pain by the nurses]. AB - The College of Nurses of Trento (Collegio IPASVI) promoted a call for research works done by nurses, to award a prize to those judged worthy. The three best works were respectively on concordance on triage classification between doctors and nurses; on the results of organizational and educational interventions on the reduction of needle-stick injuries; and on the differences on postoperative pain assessment between nurses and patients. The final reports were later discussed as part of a training exercise aiming at improving the skill of scientific writing and they are published here, together with some comments that emphasize areas of improvement in data analysis and interpretation and some broader issues related to nursing research. PMID- 9220875 TI - [Pain and the conditions of ulcers in patients with critical ischemia of the lower limbs. Gruppo i.c.a.i. (Ischemia Critica Art Inferiori)-Nursing]. AB - Aim of the study was to describe the characteristics, the strategies and the degree of control of pain and lower limb ulcers in a sample of 959 patients with c.l.i. (chronic limb ischaemia) admitted to 38 medical and surgical italian vascular wards and recruited into a multicenter controlled trial. Data presented in this article refer to the patient situation before the hospital admission. Data on the ulcer and the nursing history on ulcers medication, pain experienced and pain control were collected at hospital admission. Mean patients age is 71.4 years; men are 69.5% of the sample. 505 patients (54.4%) experience very severe pain (from 71 to 100 on a visual analogue scale) and 173 (34.3% of these patients) are not given any pain relief. Women experience more pain than men although a larger number of women are prescribed analgesics. Leg ulcers affect 647 patients (67.8%) and 47% show a necrosis, eschar or gangrene. 263 patients (46.8%) are judged to have a clinical leg ulcer infection. There is a larger use of antiseptics vs cleansing solutions (39.5 vs 12.3), and some patients use not recommended treatments: topic antibiotics (10%), antiseptic tinctures (12.5%) and alcohol soaks (4.6%), that are used irrespective of the ulcers' conditions. Major areas of improvement in the care of c.l.i. patients are related to pain control and ulcers management. PMID- 9220876 TI - [Professional and working conditions of newly licensed nurses in the province of Trento]. AB - Aim of the survey was to gather information on the occupational situation of recently trained registered nurses of the Trento Province. 484 questionnaires were mailed to all the nurses that obtained their diploma from 1993 to 1995; 379 questionnaires (78%) were returned. Among the 70 subjects not working as nurses, 19 are unemployed, 12 study full time, 26 work full time in other professions, 11 are in the army, all the others work occasionally, but not as nurses. 46% are unsatisfied or partially satisfied with their job; 87.3% would choose again the nursing profession. 20% of the nurses would prefer to work on a part time basis. PMID- 9220877 TI - [News from dermatology]. PMID- 9220879 TI - [Difficult patients--difficult physicians?]. PMID- 9220878 TI - [Rwanda and its surroundings: among displaced persons and refugees]. PMID- 9220880 TI - Outcomes. PMID- 9220881 TI - Patient education: we have a better system now. PMID- 9220882 TI - 11 neuro myths to retire now. PMID- 9220883 TI - There's always hope. PMID- 9220884 TI - Antihypertensives. PMID- 9220886 TI - Organ donors: your care is critical. PMID- 9220885 TI - Needle sticks. The ugly truth. PMID- 9220887 TI - Tuning in to the power of music. PMID- 9220888 TI - Protecting patients' privacy. PMID- 9220889 TI - A new OTC option for allergy sufferers. PMID- 9220890 TI - Killing, not caring. PMID- 9220891 TI - Education: a lifelong process for managers/leaders. PMID- 9220892 TI - A case for the MBA. PMID- 9220894 TI - Developing future managers: education and practice. AB - Selecting, educating, and training managers for the future is not optional. If you leave this to chance, then you leave the organization's future to chance. The concept of identifying personal core competencies of future managers parallels the General's challenge of the "fog of war." You cannot tell people in advance all the things they will encounter as they march the organization forward. You can only prepare them so that they can handle the unexpected and unforeseen challenges that will occur. Building into the leader the necessary personal core competency is the best preparation for surprises. The personal core competencies for our organization's work of the future are shown below: Manager's Core Competencies: Teaming, Leadership, Process Management, Systems, Business Savvy, Product Knowledge, Job Knowledge. Each core competency includes a description that paints a clear picture of our expectations of our future managers. Your competencies may have similar labels or titles, but the descriptions will be unique to the vision of the future challenges for your organization. PMID- 9220893 TI - Capitation payments and risk. PMID- 9220895 TI - Developing future nurse managers: education and practice. PMID- 9220896 TI - Reengineering knowledge and skills: essential for nurse managers. AB - Nurse managers of this decade and beyond face a very different set of performance expectations than they did only 10 years ago. Although many of the previously acquired skills remain useful, there are new skills to be assimilated to enhance organizational and clinical effectiveness. Reengineering is a valuable knowledge tool that nurse managers can put to use across health care settings. Nurse managers become process-owners, and as such can effect substantive positive change within their organizations. PMID- 9220897 TI - Community systems management: preparing nurse managers for today and tomorrow. AB - Industry changes not only demand flexibility among health care workers but also require transformation of education programs that prepare leaders and managers for practice in evolving venues of delivery. Schools of nursing are responding by listening to the market and designing curricula to meet the new demand. This article describes one cutting-edge graduate program that prepares nurses for new leadership roles in health care. PMID- 9220898 TI - New skills sets needed to navigate managed care. AB - Nurse managers in the late 1990s and the 21st century may find themselves working for a managed care organization, and certainly will find themselves employed in a managed health care environment. The roles and opportunities are many for managers who have the right combination of skills and knowledge. Understanding managed care language and strategies in synergy, with a creative leadership approach, enables nurse managers to move successfully from acute care to the new world now unfolding. PMID- 9220899 TI - Classic nursing management skills and disease management: something old, something new. AB - With the advent of managed care, nursing has the opportunity to shine through the current chaos in the health care industry because of its steadfast use and necessary expertise in the five classic management skills: planning, coordinating, controlling, delegating, and communicating. The new focus on disease management provides nursing with the best chance ever to position itself as the most skilled and pivotal player in an incredibly competitive arena. The five skills will be described in the new context of disease management, with the hope that nurses in diverse roles will demonstrate that excellent management of clinical care is a patient's best chance for achieving optimal outcomes and a health care system's best chance at achieving value. PMID- 9220900 TI - Informatics knowledge: the key to maximizing performance and productivity. AB - Nurse managers face a competitive and complex market-place that demands greater attention to customer satisfaction along with continual improvements in quality and cost-effectiveness. Without information technology, the manager cannot hope to deliver ever greater quality at less cost. The nurse managers' effective promotion and use of computer applications can have enormous impact on the performance and productivity of their health care facilities. This article explores the potential of information technology to maximize clinical and cost outcomes, optimize decision making, and enhance administrative productivity. PMID- 9220901 TI - Preparing nurse leaders for the future: views from Canada. AB - Preparing nurse managers for the present and nurse leaders for the future in the Canadian health care system is discussed from the perspective of service and education. The Canadian health care system is undergoing change, as the need to reduce and control costs comes in conflict with strongly held principles of the Canada Health Act. Nurse leaders will require knowledge and skill in transformative leadership to enable nurses to focus on the core of professional practice as clinical settings shift to self-directed, transdisciplinary teams. Nurse leaders also will need skills in building alliances within an increasingly consumer-controlled health care system, and competencies in work redesign and system design and redesign to enable nursing to smooth the "seams" in a high functioning, but not necessarily well-integrated health care system. Judgment and clarity of vision are needed to embrace conflict when necessary and to prevent the initiation of practices and structures that run counter to the values embedded in the Canada Health Act, and nursing itself. PMID- 9220902 TI - Leadership in a new health care world. AB - To understand and cope with the present, it is often very useful to examine the past. In so doing, leaders in nursing can put the events of today into perspective. Further, the rich history of nursing leadership can point us toward the future. The author draws from our history to paint a clear picture of the nurse leader of tomorrow. PMID- 9220903 TI - Health informatics. AB - This article addresses health informatics and some of the technology advancements and issues facing health care organizations today. As the ability to communicate and share data with other institutions is rapidly advancing, so is the need for industry coding standardization and data protection. PMID- 9220904 TI - Role and responsibilities of an information specialist. AB - The primary challenge in the role of an information system specialist is to combine clinical expertise with knowledge of current computer technology. Health care data must be collected, stored, and accessed in the most efficient manner possible to meet the variety of information management needs found in a health care setting (ie, patient care, research, and financial requirements. PMID- 9220906 TI - Perioperative system design and evaluation. AB - Moving toward an electronic record is both challenging and rewarding. The implementation of a computerized scheduling and management system for the operating room is evaluated. Components of the system include Scheduling, Personnel, Supply, Intraoperative, and InSight modules. PMID- 9220905 TI - Security and legal issues associated with the computerized patient record. AB - The advances in electronic technology in the form of increased processing power, the ability to computerize many aspects of a patient's record, and the emergence of document imaging systems and telemedicine have enhanced health care professionals ability to more accurately document care. However, with this augmentation comes issues in liability and security of which professionals should be aware. This article discusses some of these issues from the viewpoint of the perioperative nurse. PMID- 9220907 TI - Computers in the operating room: the staff nurse perspective. AB - Computers and information management are long-standing tools for the Perioperative Manager. As paperless nursing documentation makes its way into the operating room, the staff nurse must become adept at the use of the computer. How to get the staff nurse comfortable with this new role, and concerns the staff nurse may voice are the subject of this article. PMID- 9220909 TI - Operating room: 2010. AB - Using expert systems, virtual reality, and commercial and futuristic technology, visionary operating room (OR) nurses will have the opportunity in the 21st century to dramatically improve the way the OR functions. Eliminating counting, decreasing occurrences of patient injuries, and improving staff and patient education are just some of the possibilities! PMID- 9220910 TI - Technology: evolution or revolution--changing times. AB - Technology is changing the way work in a department is processed, often leading to greater efficiency and cost savings over time. How nurses engage in the process of business reengineering may help to determine the agency's competitive edge. This article discusses the process and the use of computer technology in the perioperative area. PMID- 9220911 TI - Use of advanced analytical techniques in assessing telemedicine. AB - Telemedicine is becoming increasingly prevalent in health care services. This article provides a general description of telemedicine and a detailed description of its use in Operation Joint Endeavor (Bosnia). Use of computer modeling for prospective analyses of telemedicine issues is focused on. Finally, this article describes the potential use of model-based analyses to address the impact of telemedicine on key nursing-related topics. PMID- 9220908 TI - Occupational health hazard: carpal tunnel syndrome. AB - A significant portion of the American population today is exposed to computer related illnesses. One of the most common injuries is carpal tunnel syndrome (CTS). Perioperative nurses will become increasingly exposed to computer-related illnesses with the advent of computerized patient record systems. Economic loss, physical disability, and emotional distress are frequent outcomes of computer related illnesses. Federal legislation addressing preventive measures is currently nonexistent. Clinicians, as both employers and consumers of computer technology, must address computer-related illnesses, such as CTS, through identification of related risk factors, early symptoms, implementation of ergonomic measures, and support of federal and industrial safety standards. PMID- 9220912 TI - Three-dimensional ultrasound and image-directed surgery: implications for operating room personnel. AB - The proliferation of new imaging technologies is having a profound impact on all surgical specialties. New means of surgical visualization are allowing more surgeries to be performed less invasively. Three-dimensional ultrasound is a technology that has potential as a diagnostic tool, as a presurgical planning simulator, and as an adjunct to image-directed surgery. This article describes how three-dimensional ultrasound is being used by the United States Department of Defense and how it may change the role of the perioperative nurse in the near future. PMID- 9220913 TI - Hundreds enrolled in canadian HIV trials. PMID- 9220914 TI - So far so good for HIV vaccine. PMID- 9220915 TI - Sex or gender? PMID- 9220916 TI - Inappropriate practices in prescribing: who decides and how? PMID- 9220917 TI - Inappropriate practices in prescribing: who decides and how. PMID- 9220918 TI - An impaired judicial system. PMID- 9220919 TI - Immunization and global ecology. PMID- 9220920 TI - Standards for polysomnography. PMID- 9220921 TI - Our future physicians deserve better. PMID- 9220922 TI - The effects of clinical practice guidelines on patient outcomes in primary care: a systematic review. AB - OBJECTIVE: To assess the evidence for the effectiveness of clinical practice guidelines (CPGs) in improving patient outcomes in primary care. DATA SOURCES: A search of the MEDLINE, HEALTHPLAN, CINAHL and FAMLI databases was conducted to identify studies published between Jan. 1, 1980, and Dec. 31, 1995, concerning the use of guidelines in primary medical care. The keywords used in the search were "clinical guidelines," "primary care," "clinical care," "intervention," "randomized controlled trial" and "effectiveness." STUDY SELECTION: Studies of the use of CPGs were selected if they involved a randomized experimental or quasi experimental method, concerned primary care, were related to clinical care and examined patient outcomes. Of 91 trials of CPGs identified through the search, 13 met the criteria for inclusion in the critical appraisal. DATA EXTRACTION: The following data were extracted, when possible, from the 13 trials: country and setting, number of physicians, number of patients (and the proportion followed to completion), length of follow-up, study method (including random assignment method), type of intervention, medical condition treated and effect on patient outcomes (including clinical and statistical significance, with confidence intervals). DATA SYNTHESIS: The most common conditions studied were hypertension (7 studies), asthma (2 studies) and cigarette smoking (2 studies). Four of the studies followed nationally developed guidelines, and 9 used locally developed guidelines. Six studies involved computerized or automated reminder systems, whereas the others relied on small-group workshops and education sessions. Only 5 of the 13 trials (38%) produced statistically significant results. CONCLUSION: There is very little evidence that the use of CPGs improves patient outcomes in primary medical care, but most studies published to date have used older guidelines and methods, which may have been insensitive to small changes in outcomes. Research is needed to determine whether the newer, evidence-based CPGs have an effect on patient outcomes. PMID- 9220923 TI - Canadian physicians' attitudes about and preferences regarding clinical practice guidelines. AB - OBJECTIVE: To assess Canadian physicians' confidence in, attitudes about and preferences regarding clinical practice guidelines. DESIGN: Cross-sectional, self administered mailed survey. PARTICIPANTS: Stratified random sample of 3000 Canadian physicians; 1878 (62.6%) responded. SETTING: Canada. OUTCOME MEASURES: Physicians' use of various information sources; familiarity with and confidence in guidelines; attitudes about guidelines and their effect on medical care; rating of importance of guidelines and other sources of information in clinical decision-making; rating of importance of various considerations in deciding whether to adopt a set of guidelines; and rating of usefulness of different formats for presenting guidelines. MAIN RESULTS: In all, 52% of the respondents reported using guidelines at least monthly, substantially less frequently than traditional information sources. Most of the respondents expressed confidence in guidelines issued by various physician organizations, but 51% to 77% were not confident in guidelines issued by federal or provincial health ministries or by health insurance plans. The respondents were generally positive about guidelines (e.g., over 50% strongly agreed that they are a convenient source of advice and good educational tools); however, 22% to 26% had concerns about loss of autonomy, the rigidity of guidelines and decreased satisfaction with medical practice. Endorsement by respected colleagues or major organizations was identified as very important by 78% and 62% of the respondents respectively in deciding whether to adopt a set of guidelines in their practice. User friendliness of the guidelines format was thought to be very important by 62%; short pamphlets, manuals summarizing a number of guidelines, journal articles and pocket cards summarizing guidelines were the preferred formats (identified as most useful by 50% to 62% of the respondents). CONCLUSIONS: Canadian physicians, although generally positive about guidelines and confident in those developed by clinicians, have not yet integrated the use of guidelines into their practices to a large extent. Our results suggest that respected organizations and opinion leaders should be involved in the development of guidelines and that the acceptability of any proposed format and medium for guidelines presentation should be pretested. PMID- 9220924 TI - Clinical practice guidelines on trial. PMID- 9220925 TI - Bioethics for clinicians: 12. Ethical dilemmas that arise in the care of pregnant women: rethinking "maternal-fetal conflicts". AB - When a pregnant woman makes a decision or acts in a manner that may be detrimental to the health and well-being of her fetus, her physician may be faced with an ethical dilemma. Is the physician's primary duty to respect the woman's autonomy, or to promote behaviour that may be in the best interest of the fetus? The controversial concept of "fetal rights" or the "fetus as a patient" contributes to the notion that the pregnant woman and her fetus are potential adversaries. However, Canadian law has upheld women's right to life, liberty and security of the person and has not recognized fetal rights. If a woman is competent and refuses medical advice, her decision must be respected even if the physician believes that her fetus will suffer as a result. Coercion of the woman is not permissible no matter what appears to be in the best interest of the fetus. PMID- 9220926 TI - For a physician with the signature scar of the CABG club, a friend's death brings not only grief but also memories and fear. PMID- 9220928 TI - For first time, unemployment line awaits group of new Canadian specialists. PMID- 9220930 TI - Dissemination of research results to clinicians an art in itself. PMID- 9220929 TI - Hospital evacuated, mental-health issues dominated as Manitoba coped with flood of century. PMID- 9220927 TI - Viral hepatitis: know your D, E, F and Gs. PMID- 9220931 TI - Are the lean days for research funding finally coming to an end? PMID- 9220932 TI - Lessons from Amy. PMID- 9220933 TI - Lessons from Amy. PMID- 9220934 TI - Another look at those OECD numbers. PMID- 9220935 TI - Debate over dexfenfluramine. PMID- 9220936 TI - Alert over sound-alike drugs. PMID- 9220937 TI - When to scan, when to operate. PMID- 9220939 TI - Does good science make good medicine? Incorporating evidence into practice is complicated by the fact that clinical practice is as much art as science. PMID- 9220938 TI - Contemporary practice patterns in the management of newly diagnosed hypertension. AB - OBJECTIVE: To determine what proportion of patients with hypertension are managed in accordance with guidelines established by the Canadian Hypertension Society. DESIGN: Retrospective medical record review. SETTING: Outpatients seen in primary care offices and internal medicine referral clinics in Edmonton. PATIENTS: All 969 adults who presented with a new diagnosis of essential hypertension from Sept. 1, 1993, to Dec. 31, 1995. OUTCOME MEASURES: Initial laboratory tests performed, advice concerning nonpharmacologic treatment given, antihypertensive drugs prescribed and any contraindications to thiazide diuretics or beta adrenergic blocking agents documented. RESULTS: The mean age of the 969 patients in the sample was 52.5 years; 129 (13%) of the patients were older than 70 years of age; and 500 (52%) were women. Most of the patients (704, 73%) had mild or moderate diastolic hypertension. In the 617 patients who underwent laboratory tests related to hypertension, the creatinine level was determined in 466 (76%), the cholesterol level in 372 (60%), a urinalysis was conducted in 378 (61%), the serum potassium level was checked in 343 (56%), the sodium level in 323 (52%) and an electrocardiogram was performed in 303 (49%). Liver function tests, which are not recommended in the guidelines, were performed in 338 patients (55%). Although there were differences in prescribing among physicians in the 711 patients given first-line therapy, most (238, 34%) were prescribed angiotensin-converting-enzyme (ACE) inhibitors. Lifestyle modification, without drug therapy, was suggested for 180 (25%) of the patients. Although the guidelines recommend their use for first line drug therapy, only 82 patients (12%) were given beta-adrenergic blocking agents and only 75 (11%) were given thiazide diuretics. Of the patients who were prescribed an antihypertensive other than a thiazide or beta-adrenergic blocking agent as first-line drug therapy, only 161 (43%) had a documented contraindication to thiazides or beta-adrenergic blocking agents. CONCLUSIONS: There is variation in the contemporary care of patients with hypertension. Further studies are required to determine the reasons underlying physicians' noncompliance with the evidence-based guidelines established by the Canadian Hypertension Society. PMID- 9220940 TI - Terfenadine therapy: can we justify the risks? PMID- 9220941 TI - The times they are confusing. What lies ahead for the new health minister and physicians in Canada? PMID- 9220942 TI - Primary care reform: is it time for population-based funding? PMID- 9220943 TI - A new primary care rostering and capitation system in Norway: lessons for Canada? AB - Providing every patient with a personal primary care physician or, from the physician's perspective, establishing a stable roster or list of patients is currently being actively debated in Canada. Norway's system of primary care medicine, similar to Canada's, faces many of the same problems. In 1992 a trial rostering system with blended funding (capitation, fee-for-service and user fees) was established in 4 Norwegian municipalities. After 3 years of close monitoring, the results of system evaluations have attracted strong interest. This article reports on the benefits and problems encountered with the new rostering system in Norway. If Canada is moving in the same direction, some of the lessons learned may be helpful. PMID- 9220944 TI - Bat rabies after undetected exposure: implications for prophylaxis. PMID- 9220945 TI - Health care among forgotten issues in forgettable federal election. PMID- 9220946 TI - Fetal alcohol syndrome epidemic on Manitoba reserve. PMID- 9220947 TI - "Ask the gambling question," FPs told as "secret" addiction becomes more common. AB - Family physicians were recently advised on ways to determine if their patients are addicted to gambling. Jim Milligan of Toronto's Donwood Institute says the problem is becoming more common but is often difficult to detect. PMID- 9220948 TI - Obesity in Canada: a descriptive analysis. Canadian Heart Health Surveys Research Group. AB - OBJECTIVE: To describe the distribution of body fat, prevalence of obesity, and knowledge of cardiovascular disease in Canadian adults. DESIGN: Population-based, cross-sectional surveys. SETTING: Ten Canadian provinces between 1986 and 1992. PARTICIPANTS: A probability sample of 29,855 men and women aged 18 to 74 years was selected using health insurance registration files in each province. Anthropometry was performed on 19,841 (66%) of these adults. OUTCOME MEASURES: Body mass index (BMI); waist circumference; ratio of waist to hip circumference; knowledge of causes of heart disease. RESULTS: The overall prevalence of obesity (BMI > or = 27 kg/m2) increased with age and was greater in men (35%) than in women (27%). Abdominal obesity was also higher in men and increased with both age and BMI. Canadians with lower levels of education had a higher prevalence of obesity, which appeared at a young age. Canadians in Atlantic Canada mentioned lack of exercise, poor diet and smoking as causes of heart disease less frequently than those living in central or western Canada. CONCLUSIONS: Obesity continues to be common among Canadian adults. Policy and programs to promote healthy body weights must be intensified and directed at specific sociodemographic groups. PMID- 9220949 TI - Regional and rural-urban differences in obesity in Canada. Canadian Heart Health Surveys Research Group. AB - OBJECTIVE: To describe regional and rural-urban differences in weight and weight loss patterns in Canadian adults. DESIGN: Population-based, cross-sectional surveys. SETTING: Nine Canadian provinces (excluding Nova Scotia) from 1986 to 1992. PARTICIPANTS: A probability sample of 27,120 men and women aged 18 to 74 years was selected using the health insurance registration files in each province. Anthropometry was performed on 18,043 participants (67%). OUTCOME MEASURES: Region of Canada (Atlantic, central, western); rural or urban residence (rural if participant resided in a community whose population was < 10,000, urban if population > or = 10,000); body mass index (BMI, kg/m2); percentage of participants trying to lose weight; reasons for trying to lose weight; level of leisure-time physical activity. RESULTS: Overall, mean BMI values in rural men (26.1 kg/m2) and women (25.3 kg/m2) were not significantly different from urban counterparts (25.7 kg/m2 and 24.8 kg/m2, respectively). Similarly, obesity (BMI > or = 27 kg/m2) was as prevalent in rural men (37%) and women (30%) as in urban participants (34% and 28%, respectively). However, a difference was observed in western Canada where 41% of rural and 34% of urban men were obese (odds ratio [OR], adjusted for age and education = 1.29; 95% confidence interval [CI] 1.06, 1.57), as were 35% of rural and 25% of urban women (OR, adjusted for age and education = 1.47; 95% CI 1.17, 1.84). Among men in western Canada, the rural urban differences were greatest in the 25-64 year age group, whereas in women the differences were present at all ages. Overall, in Canada, urban men (26%) are more likely than rural men (23%) to be trying to lose weight; the reverse was true for women (39% and 42%, respectively). CONCLUSION: Considerable regional and rural-urban differences are seen in the patterns of weight and weight loss in Canada. A fuller understanding of the underlying behavioural determinants of these differences is needed. On the basis of such an understanding, effective programs to promote healthy weights for individuals and communities in these areas might be developed. PMID- 9220950 TI - Weight dissatisfaction and weight loss attempts among Canadian adults. Canadian Heart Health Surveys Research Group. AB - OBJECTIVE: To describe the pattern of weight dissatisfaction and weight loss attempts among Canadian adults and the reasons for and methods of weight loss among those trying to lose weight. DESIGN: Population-based, cross-sectional surveys. SETTING: Ten Canadian provinces between 1986 and 1992. PARTICIPANTS: A probability sample of 29,855 men and women aged 18 to 74 years was selected using provincial health insurance registration files; this paper describes the subsample of 19,841 (66%) participants from whom anthropometric data were collected. OUTCOME MEASURES: Discrepancy between actual and desired body mass index (BMI); attempts to lose weight; reasons for losing weight; methods of weight loss used. RESULTS: Whether their weight was in the acceptable range (BMI 20-24 kg/m2) or at a level of increasing risk (BMI > or = 27 kg/m2), women were more likely than men to wish they weighed less and to be trying to lose weight; almost two-thirds of women but less than half the men with BMI > or = 27 kg/m2 were trying to lose weight. Even among those with BMI 20-24 kg/m2, 32% of women (v. 10% of men) were trying to reduce their weight. Weight dissatisfaction and current and past weight loss attempts were all negatively associated with age among women, but were unrelated to age among men. People with higher ratios of waist to hip circumference (WHR), controlling for BMI, were no more likely to be trying to lose weight than those with lower WHR; in fact, for women with BMI 27 29 kg/m2, WHR was negatively associated with prevalence of weight loss attempts. The presence of diabetes, hypertension and hypercholesterolemia was also unrelated to weight loss attempts; regular smokers and sedentary people were less likely to report trying to lose weight, controlling for BMI. Among those currently trying to lose weight, the most commonly mentioned reason was to improve general health, followed by increasing attractiveness. Overall, the most frequently mentioned method of weight loss was dieting, followed by exercise. CONCLUSIONS: Substantial numbers of men whose BMI places them at increased health risk appear to be content with their weight and are not attempting to reduce it. Conversely, women, especially the young and middle-aged, are likely to consider themselves above their desired weight and to be trying to lose weight, even when their weight is within acceptable limits. This reinforces the need to consider differences between men and women in efforts to promote and support healthy weights among Canadians. PMID- 9220951 TI - Risk factor correlates of body mass index. Canadian Heart Health Surveys Research Group. AB - OBJECTIVE: To examine the association of obesity, as reflected by body mass index, with other cardiovascular risk factors specifically blood pressure, smoking, physical inactivity, plasma lipid levels and diabetes mellitus. DESIGN: Population-based, cross-sectional surveys. SETTING: Ten Canadian provinces between 1986 and 1992. PARTICIPANTS: A probability sample of 29,855 men and women aged 18 to 74 years was selected from the health insurance registration files of each province and invited to participate. Anthropometry was performed on 19,841 (66%) of these adults. OUTCOME MEASURES: Body mass index (BMI, kg/m2), systolic and diastolic blood pressure, smoking status, level of leisure-time physical activity, self-reported diabetes, levels of plasma total cholesterol, high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL) and triglycerides (TRIG). RESULTS: The prevalence of high blood pressure increased with increasing BMI. The gradient of increase was steepest for younger (18-34 years) men and women compared with older (55-74 years) groups. The prevalence of physical inactivity in women tended to increase with increasing BMI except in the lowest BMI category. The J-shaped relationship, although weaker, was also seen in men. The prevalence of self-reported diabetes mellitus was greater with higher BMI categories at all ages and for both sexes except for the youngest group of men. The prevalence of dyslipidemia was related to BMI, as LDL and TRIG levels were higher and HDL levels lower in those with higher BMI. BMI was strongly related to blood pressure, diabetes mellitus and lipid abnormalities. CONCLUSION: These data suggest a central role for obesity in cardiovascular risk and the potential importance of intervention strategies aimed at reducing population obesity in the management of other cardiovascular risk factors. PMID- 9220952 TI - A comparative analysis of weight to height and waist to hip circumference indices as indicators of the presence of cardiovascular disease risk factors. Canadian Heart Health Surveys Research Group. AB - OBJECTIVE: To determine the mathematic formula for weight, height and waist and hip circumference that is most closely correlated to cardiovascular disease risk factors. DESIGN: Population-based, cross-sectional surveys. SETTING: Five Canadian provinces, between 1990 and 1992. PARTICIPANTS: A probability sample of 16,007 men and women aged 18 to 74 years was selected using health insurance registration files in each province. Anthropometry was performed on 10,054 (63%) of these adults. OUTCOME MEASURES: The power of height in the body mass index (BMI, kg/m2) and of hip circumference in the ratio of waist to hip circumference (WC/HC) was varied from 0 to 3. Simple linear regression analysis for each age sex group was used to examine the relation of each index to systolic and diastolic blood pressure (SBP and DBP), levels of total cholesterol (TC), low density lipoprotein cholesterol (LDL) and high-density lipoprotein cholesterol (HDL), triglycerides (TRIG) and the ratio of TC to HDL. Values for the coefficient of determination (r2) were used to compare the fits of the models. RESULTS: The r2 values were generally low (< 0.27), but were greatest in the younger age groups (18-24 and 35-54 years) and in women. Waist circumference alone (WC/HC0) showed the best fit with SBP and DBP, whereas WC/HC0.5 was most closely related to HDL, TC/HDL and TRIG. None of the indices was closely associated with TC or LDL. Whatever the power of height used, the weight-height ratios showed weaker associations with the risk factors than the waist-hip ratios. CONCLUSIONS: WC and BMI correlate most closely with blood pressure and plasma lipid and may be the best simple anthropometric indices to include in the routine clinical examination of adults. PMID- 9220953 TI - The association of cardiovascular disease risk factors with abdominal obesity in Canada. Canadian Heart Health Surveys Research Group. AB - OBJECTIVE: To assess the degree of association of abdominal obesity with blood pressure and plasma lipid levels and to determine which anthropometric measures of obesity are most closely associated with these cardiovascular risk factors. DESIGN: Population-based, cross-sectional surveys. SETTING: Five Canadian provinces (Alberta, Manitoba, Ontario, Quebec and Saskatchewan) between 1989 and 1992. PARTICIPANTS: A probability sample of 16,007 men and women aged 18 to 74 was selected using health insurance registration files in each province and invited to participate. A complete set of measurements was available for 8974 (56%) adults. OUTCOME MEASURES: Initially, simple correlation analyses by age and sex were performed between the anthropometric variables-body mass index, waist circumference (WC), hip circumference (HC), ratio of waist to hip circumference (WHR)- and cardiovascular disease risk variables-systolic blood pressure (SBP), diastolic blood pressure (DBP), levels of total cholesterol (TC), low density lipoprotein (LDL) cholesterol, high density lipoprotein (HDL) cholesterol and triglycerides (TRIG) and the TC/HDL ratio. Canonical correlation analyses were performed to determine the multivariate associations between the anthropometric and risk variables. RESULTS: The simple correlations between anthropometric variables and cardiovascular disease risk variables were highest for SBP; moderate for DBP, HDL, TRIG and TC/HDL; and lowest for LDL and TC. Of the anthropometric variables, WC demonstrated the greatest correlations with the risk variables. The first canonical correlations were significant (p < 0.0001) in men (0.58) and women (0.61) of all ages. Of the anthropometric variables, WC consistently demonstrated the highest loading values in the first canonical variable in men (0.56) and women (0.59). Of the risk variables in both sexes, the loadings of TRIG were generally the largest, those of HDL, SBP, DBP intermediate and those of LDL the smallest. In men, the strength of these associations generally decreased with age, whereas in women they peaked in the 35-54 year age group. CONCLUSION: Considerable association was seen between measures of abdominal obesity and blood pressure and plasma lipid levels. WC is the measure of abdominal obesity most highly correlated with these cardiovascular disease risk factors. PMID- 9220954 TI - Correlation between cardiovascular disease risk factors and simple anthropometric measures. Canadian Heart Health Surveys Research Group. AB - OBJECTIVE: To assess simple anthropometric measures as indicators of the concurrent presence of high blood pressure, dyslipidemia and diabetes mellitus in adults. DESIGN: Population-based, cross-sectional surveys. SETTING: Five Canadian provinces between 1990 and 1992. PARTICIPANTS: A probability sample of 16,007 men and women aged 18 to 74 years was selected using health insurance registration files in each province. This study is based on the 9826 adults (61%) for whom anthropometric measurements were obtained. OUTCOME MEASURES: Step-wise multiple logistic regression analysis was used to model the association between demographic, anthropometric and risk variables and the presence of high systolic and diastolic (DBP) blood pressure, elevated levels of total (TC), high-density lipoprotein (HDL) and low-density lipoprotein cholesterol, TC/HDL ratio, triglyceride levels (TRIG) and self-reported diabetes mellitus. RESULTS: Age group and sex are strongly associated with all three conditions. Sedentary lifestyle is significantly associated with high DBP, depressed HDL and elevated TC/HDL and TRIG. Anthropometric measures are moderately associated with all conditions. The measures of body fat (body mass index) as well as abdominal fat distribution (waist circumference and ratio of waist to hip circumference) play an approximately equal role. CONCLUSION: Patients' age, sex, level of physical activity, body fat and abdominal fat distribution can be used as indicators of the probability of high blood pressure, dyslipidemia and diabetes mellitus. PMID- 9220955 TI - Effects of neutralization pattern and stereochemistry on DNA bending by methylphosphonate substitutions. AB - Asymmetric phosphate neutralization has been hypothesized to play a role in DNA bending by proteins. Neutralization is thought to involve salt bridges between the negatively charged phosphate backbone of duplex DNA and the cationic amino acids of an approaching protein. According to this model, the resulting unbalanced charge distribution along the duplex DNA induces the double helix to collapse toward the neutralized surface. Previous work has confirmed that DNA bending is induced by the asymmetric incorporation of racemic methylphosphonate linkages creating a neutral region on one face of duplex DNA. Neutralization was accomplished by substitution of three consecutive phosphodiesters on each strand, arranged across one minor groove of the DNA (a total of six neutralized phosphates). We now measure DNA bending induced by a more diffuse patch of neutralization (alternating neutralized and anionic phosphates) and explore the effect of methylphosphonate stereochemistry. DNA duplexes with patches of alternating methylphosphonate and phosphodiester linkages are less bent than DNAs wherein consecutive phosphates are neutralized. Furthermore, duplexes neutralized by incorporation of pure (RP)-methylphosphonate isomers are bent approximately 30% less than duplexes neutralized by racemic methylphosphonates. PMID- 9220956 TI - Interaction of tRNA with tRNA (guanosine-1)methyltransferase: binding specificity determinants involve the dinucleotide G36pG37 and tertiary structure. AB - The sequence G37pG36 is present in all tRNA species recognized and methylated by the Escherichia coli modification enzyme tRNA (guanosine-1)methyltransferase. We have examined whether this dinucleotide sequence provides the base specific recognition signal for this enzyme and have assessed the role of the remaining tRNA in recognition. E. coli tRNAHis and yeast tRNAAsp were substituted with G at positions 36 and 37 and were found to be excellent substrates for methylation. This suggested that the general tRNA structure can be specifically bound by the enzyme. In addition, heterologous tRNA species including fully modified tRNA1Leu are excellent inhibitors of tRNA1Leu transcript methylation. Analyses of structural variants of yeast tRNAAsp and E. coli tRNA1Leu demonstrate clearly that the core tertiary structures of tRNA are required for recognition and that G37 must be in the correct position in space relative to important contacts elsewhere in the molecule. This latter conclusion was reached because the addition of one to three stacked base pairs in the anticodon stem of tRNA1Leu dramatically alters activity. In this case, the G37 base is rotated away from the correct position in space relative to other tRNA contact sites. The acceptor stem structure is required for optimal activity since deletion of three or five base pairs is detrimental to activity; however, specific base sequence may not be important because (i) the addition of three stacked base pairs of different sequence had little effect on activity and (ii) heterologous tRNAs with little or no sequence homology in the acceptor stem are excellent substrates. Both poly G and GpG are potent and specific inhibitors of enzyme activity and are minimal substrates which can be methylated, forming m1G. Taken together, these studies suggest that 1MGT can bind the general tRNA structure and that the crucial base pair contacts are G37 and G36. PMID- 9220957 TI - Gem-dialkyl succinic acids: a novel class of inhibitors for carboxypeptidases. AB - gem-Dimethylsuccinic acid and its higher homolog, 2-methyl-2-ethylsuccinic acid (MESA) are highly potent inhibitors of both carboxypeptidase A (CPA) and B. The inhibition constant of MESA for CPA (0.11 microM for the racemic mixture) is remarkable considering the relatively simple structure of the compound. The molecular feature which is crucial for high affinity binding to both carboxypeptidases appears to be the nonpolar gem-dialkyl locus. The structure of the complex between MESA and CPA has been determined by X-ray crystallography to 2.0 A resolution and shows the R enantiomer of the inhibitor to be bound in a generally substrate-like manner. The carboxymethyl group is coordinated to the Zn ion in the active site, and the gem-dialkyl locus corresponds in position to the alpha-carbon of the C-terminal amino acid in a peptide substrate. The methyl group of the inhibitor occupies a cavity in the enzyme which is apparently not filled upon substrate-binding. We postulate that this cavity (the alpha-methyl hole) is designed to allow the proximal Glu-270 residue to undergo a critical movement during catalysis. The hydrophobic nature of the above cavity may play a role in modulating the reactivity of this residue. These results suggest that similar cenophilic(empty-loving) inhibitors may be found for other enzymes. PMID- 9220958 TI - Collagen-based structures containing the peptoid residue N-isobutylglycine (Nleu): synthesis and biophysical studies of Gly-Nleu-Pro sequences by circular dichroism and optical rotation. AB - Single-chain peptide-peptoid structures, Ac-(Gly-Nleu-Pro)n-NH2 (n = 3, 6, and 10) and (Gly-Nleu-Pro)n-NH2 (n = 1 and 9), and template-assembled collagen analogs, KTA-[Gly-(Gly-Nleu-Pro)n-NH2]3 (n = 3 and 6; KTA represents cis,cis 1,3,5-trimethylcyclohexane-1,3, 5-tricarboxylic acid, also known as the Kemp triacid; Nleu denotes N-isobutylglycine), were prepared by solid-phase peptide synthesis methods. Biophysical studies using circular dichroism (CD) and optical rotation measurements show that these collagen analogs form triple-helical conformations when the chain is longer than a critical length. Unlike collagen based structures composed of Gly-Pro-Hyp and Gly-Pro-Nleu sequences, results reveal that the presence of a positive CD peak between 220 and 225 nm is indicative of triple-helical conformations for these collagen-based structures composed of Gly-Nleu-Pro sequences. Results also indicate that the Gly-Nleu-Pro sequence possesses a higher triple-helical propensity than the Gly-Pro-Nleu sequence as demonstrated by the higher melting temperatures, the faster triple helix folding, and the lower minimum concentration necessary to detect triple helicity for the single-chain structures. Therefore, we conclude that the Nleu residue in the second position of the trimeric repeat is more effective in inducing triple-helix formation than Pro in the same position. PMID- 9220959 TI - Collagen-based structures containing the peptoid residue N-isobutylglycine (Nleu): conformational analysis of Gly-Nleu-Pro sequences by 1H-NMR and molecular modeling. AB - Molecular modeling and 1H-NMR were employed to study the structure and stability of collagen-like triple helices composed of Gly-Nleu-Pro repeats. The compounds studied include the acetyl analogs Ac-(Gly-Nleu-Pro)n-NH2 (where n = 1, 3, 6, and 10) and the KTA conjugates KTA-[Gly-(Gly-Nleu-Pro)n-NH2]3 (where n = 3 and 6 and KTA denotes the Kemp triacid). The presence of collagen-like assembled structures is supported by a consistent set of experimental observations, which include the appearance of a distinct set of resonances, low hydrogen-exchange rates for Gly NH, cooperative melting transition, and observation of several interchain NOEs. Using 1H-NMR, the triple helicity was monitored as a function of chain length, template, and temperature. These studies show that (Gly-Nleu-Pro)n sequences have a somewhat higher triple-helical propensity than (Gly-Pro-Nleu)n sequences. In addition, our investigations have shown that unlike the triple helices composed of Gly-Pro-Nleu repeats those composed of Gly-Nleu-Pro repeats can access conformations in which the Nleu side chains are arrayed between Pro residues belonging to different triple-helix cross sections. These structural features may serve as a basis for free energy computations and for the study of higher-order structures such as collagen-like fibrils containing peptoid moities. PMID- 9220960 TI - The role of phenylalanine 31 in maintaining the conformational stability of ribonuclease P2 from Sulfolobus solfataricus under extreme conditions of temperature and pressure. AB - Ribonuclease P2 from the thermophilic archaebacterium Sulfolobus solfataricus is a small protein (7 kDa) with a known three-dimensional structure. Inspection of the structure and molecular dynamics simulation reveal that three aromatic residues (Phe5, Phe31, and Tyr33) from the hydrophobic core have a strong van der Waals interaction energy. We studied the thermodynamics of the heat, cold, and pressure-induced protein conformational changes of the wild type and of the F31A and F31Y mutants by analyzing the protein UV absorbance in the fourth derivative mode. The wild-type protein was extremely stable under all conditions of temperature and pressure. Heat and cold denaturation of both mutants, as well as denaturation by pressure of the F31A mutant, led to significant blue shifts of the derivative spectrum, indicating increased solvent exposure of Tyr33. For the F31Y mutant, high pressure (400 MPa) protected the protein against thermal denaturation. This study, probing the properties of the hydrophobic aromatic core, complements a thermal unfolding study which probes the overall structural changes [Knapp, S., Karshikoff, A., Berndt, K. D., Christova, P., Atanasov, B., & Ladenstein, R. (1996) J. Mol. Biol. 264,1132-1144]. The differences observed in response to extremes of temperature, pressure, and pH may be rationalized by an unfolding mechanism involving larger parts of the peripheral protein while the integrity of the hydrophobic core is maintained. PMID- 9220961 TI - Electrostatic effects in the kinetics of coenzyme binding to isozymes of alcohol dehydrogenase from horse liver. AB - The kinetic mechanism for the binding of NAD+ and NADH to the EE and SS isozymes of alcohol dehydrogenase (LADH) was studied between pH 7 and pH 10 by monitoring the quenching of tryptophan fluorescence. A consistent interpretation of all data was only possible by introducing a two-step binding mechanism. The first binding step is related to docking of the adenosine part of the coenzymes and the subsequent isomerization to the binding of the nicotinamide part. At high NADH concentrations an additional slow isomerization was identified as a conformational transition of the protein. A pH dependence for NADH binding is observed which is restricted to changes in the binding kinetics of the adenosine moiety going from pH 7 to pH 10, a tendency which is similar also for NAD+. This is attributed to pH-dependent variations in electrostatic attractions acting as a steering force of the docking process. The nicotinamide docking of NADH is equally fast for both isozymes and pH-independent over the measured range, whereas this docking equilibrium for NAD+ is pH-dependent for EE- and SS-LADH alike and the rate of association comparable. Presumably, a GluEE-366-LysSS substitution results in a stronger binding and faster association of both oxidized and reduced cofactor to the SS isozyme. A structural proof is presented for coenzyme-competitive binding of a sulfate ion, resulting in electrostatic shielding. PMID- 9220962 TI - Definition of the switch surface in the solution structure of Cdc42Hs. AB - Proteins of the rho subfamily of ras GTPases have been shown to be crucial components of pathways leading to cell growth and the establishment of cell polarity and mobility. Presented here is the solution structure of one such protein, Cdc42Hs, which provides insight into the structural basis for specificity of interactions between this protein and its effector and regulatory proteins. Standard heteronuclear NMR methods were used to assign the protein, and approximately 2100 distance and dihedral angle constraints were used to calculate a set of 20 structures using a combination of distance geometry and simulated annealing refinement. These structures show overall similarity to those of other GTP-binding proteins, with some exceptions. The regions corresponding to switch I and switch II in H-ras are disordered, and no evidence was found for an alpha helix in switch II. The 13-residue insertion, which is only present in rho subtype proteins and has been shown to be an important mediator of binding of regulatory and target proteins, forms a compact structure containing a short helix lying adjacent to the beta4-alpha3 loop. The insert forms one edge of a "switch surface" and, unexpectedly, does not change conformation upon activation of the protein by the exchange of GTP analogs for GDP. These studies indicate the insert region forms a stable invariant "footrest" for docking of regulatory and effector proteins. PMID- 9220963 TI - Circularly permuted beta-lactamase from Staphylococcus aureus PC1. AB - The role that domain flexibility plays in the enzymatic activity of beta lactamase from Staphylococcus aureus PC1 was investigated by producing two circularly permuted molecules. The C- and N-termini of the wild-type enzyme are adjacent to each other and remote from the active site, which is located between two domains. The polypeptide chain crosses over from one domain to the other twice. For the circularly permuted molecules, the termini were joined by an eight amino acid residue insertion, and new termini were introduced elsewhere. The first construct, termed cp254, was cleaved in a loop remote from the domain interface. The crystal structure of cp254 has been determined and refined at 1.8 A resolution, revealing essentially the same structure as that of the native protein. The activity profile with a representative sample of beta-lactam antibiotics is also very similar to that of wild-type beta-lactamase. The termini of the second circularly permuted mutant, cp228, occur within the second crossover region and therefore may enhance the flexibility of the molecule. Cp228 beta-lactamase shows a large decrease in enzymatic activity toward the sample of beta-lactam antibiotics, with catalytic rates that are 0.5-1% of those of the wild-type enzyme. One exception is the hydrolysis of the third generation cephalosporin, cefotaxime, which is hydrolyzed by the cp228 enzyme 10-fold faster than by wild-type beta-lactamase. Cp228 has not been crystallized. However, the circular dichroism spectra of the two circularly permuted proteins are very similar, indicating that, by analogy to cp254, cp228 adopts a global folded state. Thermal denaturation experiments reveal that cp254 is somewhat less stable than the wild-type enzyme, whereas cp228 is substantially less stable. Together, the data highlight the profound consequences that introducing flexibility at the domain interface has on both enzyme activity and protein stability. PMID- 9220965 TI - Existence of two distinct aspartyl-tRNA synthetases in Thermus thermophilus. Structural and biochemical properties of the two enzymes. AB - Two aspartyl-tRNA synthetases (AspRSs) were isolated from Thermus thermophilus HB8. Both are alpha2 dimers but differ in the length of their polypeptide chains (AspRS1, 68 kDa; and AspRS2, 51 kDa). Both chains start with Met and are deprived of common sequences to a significant extent. This rules out the possibility that AspRS2 is derived from AspRS1 by proteolysis, in agreement with specific recognition of each AspRS by the homologous antibodies. DNA probes derived from N terminal amino acid sequences hybridize specifically to different genomic DNA fragments, revealing that the two AspRSs are encoded by distinct genes. Both enzymes are present in various strains from T. thermophilus and along the growth cycle of the bacteria, suggesting that they are constitutive. Kinetic investigations show that the two enzymes are specific for aspartic acid activation and tRNAAsp charging. tRNA aspartylation by the thermostable AspRSs is governed by thermodynamic parameters which values are similar to those measured for mesophilic aspartylation systems. Both thermophilic AspRSs are deprived of species specificity for tRNA aspartylation and exhibit N-terminal sequence signatures found in other AspRSs, suggesting that they are evolutionarily related to AspRSs from mesophilic prokaryotes and eukaryotes. Comparison of the efficiency of tRNA aspartylation by each enzyme under conditions approaching the physiological ones suggests that in vivo tRNAAsp charging is essentially ensured by AspRS1, although AspRS2 is the major species. The physiological significance of the two different AspRSs in T. thermophilus is discussed. PMID- 9220964 TI - Chemical mechanism of the fructose-6-phosphate,2-kinase reaction from the pH dependence of kinetic parameters of site-directed mutants of active site basic residues. AB - A bifunctional enzyme, fructose-6-phosphate 2-kinase-fructose 2, 6 bisphosphatase, catalyzes synthesis and degradation of fructose 2, 6 bisphosphate. Mutants of basic residues, including Lys51, Arg78, Arg79, Arg136, Lys172, and Arg193, immediately around the active site of rat testis fructose 6 P,2-kinase were constructed, and their steady state kinetics, ATP binding, and the effect of pH on the kinetics were characterized. All mutants showed a several fold increase in KMgATP, much larger increases in KFru 6-P, and decreased V compared to those of the wild type enzyme (WT). Replacement of Lys172 and Arg193 with Ala and Leu, respectively, also produced mutants with large KFru 6-P values. Substitution of Lys51, which is located in a Walker-A motif (GXXGXGKT, amino acids 45-52), with Ala or His resulted in enzymes with increased KMgATP values and unable to bind Fru 6-P. The dissociation constants for 2'(3')-O-(N methylanthraniloyl)-ATP (mantATP) and ATP of all these mutants except Lys51 were similar. Lys51 mutants were unable to bind mantATP. The pH dependence of V and the V/Ks for MgATP and Fru 6-P suggest a mechanism in which reactants and enzyme combine irrespective of the protonation state of groups required for binding and catalysis, but only the correctly protonated enzyme-substrate complex is catalytically active. A chemical mechanism is suggested in which a general base accepts a proton from the 2-hydroxyl of Fru 6-P concomitant with nucleophilic attack on the gamma-phosphate of MgATP. Phosphoryl transfer is also facilitated by interaction of the gamma-phosphate with a positively charged residue that neutralizes the remaining negative charge. The dianionic form of the 6-phosphate of fructose 6-P is required for binding, and it is likely anchored by a positively charged enzyme residue. A comparison of the pH dependence of kinetic parameters for Ala or His mutant proteins at Lys51, Lys172, and Arg79 suggests that Lys51 interacts with the gamma-phosphate of MgATP and that several other arginines likely participate in transition state stabilization of the transferred phosphoryl. The active site general base has yet to be identified. PMID- 9220966 TI - L-delta-(alpha-Aminoadipoyl)-L-cysteinyl-D-valine synthetase: thioesterification of valine is not obligatory for peptide bond formation. AB - L-delta-(alpha-Aminoadipoyl)-L-cysteinyl-D-valine (ACV) synthetase is probably the simplest known peptide synthetase in terms of the number of reactions catalyzed. In the "thiol-template" proposal for nonribosomal peptide synthesis, a key step is transfer of aminoacyl groups derived from the substrates to enzyme bound thiols prior to peptide bond formation. No incorporation of 18O was seen in AMP isolated from the reaction mixture when di[18O]valine was incubated with relatively large amounts of active synthetase and MgATP. We therefore utilized di[18O]valine as a substrate for the biosynthesis of the diastereomeric dipeptides L-O-(methylserinyl)-L-valine and L-O-(methylserinyl)-D-valine [Shiau, C.-Y., Baldwin, J. E., Byford, M. F., Sobey, W. J., & Schofield, C. J. (1995) FEBS Lett. 358, 97-100]. In the L-O-(methylserinyl)-L-valine product, no significant loss of 18O was observed. However, in the L-O-(methylserinyl)-D valine product, a significant loss of one or both 18O labels was observed. Thus, both peptide bond formation and the epimerization of the valine residue can both occur before formation of any thioester bond to the valine carboxylate in the biosynthesis of these dipeptides. The usual qualitative test for thioesterification of substrates to the synthetase, lability of enzyme-bound radiolabeled amino acid to performic acid, proved inconclusive in our hands. These results require a new mechanism for the enzymic synthesis of L-O (methylserinyl)-L-valine and L-O-(methylserinyl)-D-valine and imply that a revised mechanism for ACV synthesis is also required. PMID- 9220967 TI - Productive interactions between the two domains of pig heart CoA transferase during folding and assembly. AB - The enzyme CoA transferase from porcine heart (EC 2.8.3.5) is a homodimer; each subunit consists of two domains linked by a hydrophilic "hinge" region. We have prepared separate DNA segments encoding each of these domains. Incorporation of these two DNA segments within an operon or within two separate transcription units does not preclude the synthesis and assembly of CoA transferase in Escherichia coli. When the two domain fragments are produced and purified individually from separate cultures and subsequently mixed, enzyme activity accumulates to near wild-type levels only after a lengthy incubation. Each domain is more susceptible to aggregation than wild-type CoA transferase. Circular dichroism shows that, prior to mixing, the domains possess a different secondary structural profile compared to their counterparts in the native enzyme. However, mixing and incubation of the domains produces a complex with far-UV CD, fluorescence, and ultracentrifugation properties similar to those of wild-type CoA transferase. Finally, we show that the intact hydrophilic peptide which links the two domains is essential for the recovery of activity observed upon refolding of the denatured enzyme in vitro. These results indicate that the folding and assembly of pig heart CoA transferase require a productive interaction between its two domains, involving a substantial conformational rearrangement. PMID- 9220968 TI - A recombinant form of the human BP180 ectodomain forms a collagen-like homotrimeric complex. AB - BP180 is a glycoprotein constituent of the epidermal anchoring complex and a major antigenic target of autoantibodies associated with bullous pemphigoid, a blistering skin disease. The C-terminal extracellular domain of BP180 contains 15 domains composed of Gly-X-Y tandem repeats, which are predicted to form collagen like triple helices. To facilitate the structural analysis of this protein, the extracellular region of human BP180 was expressed as a secreted protein (sec180e) in transiently transfected COS-1 cells. Gel filtration and sedimentation analyses demonstrated that sec180e exists in two forms: a globular monomeric form and a high-molecular mass multimeric form with an elongated conformation. Pulse-chase and cross-linking experiments established that the sec180e complex is a stable homotrimeric structure which assembles prior to secretion from the cell. On the basis of its calculated molecular mass, the oligomeric state of the sec180e complex is 3.25. With a Stokes radius of 13.6 nm, a sedimentation coefficent of 6.5 S, and a frictional ratio of 3.01, the sec180e protein appears to be highly extended (length to width ratio is between 52 and 60), yet is more flexible than a rigid rod. BP180 isolated from human epidermis was also shown to exist in a high-molecular mass complex which, like sec180e and other collagenous proteins, is SDS-stable but heat-labile. These findings strongly suggest that the BP180 ectodomain exists as an elongate, flexible homotrimer. This trimerization is likely to result from the formation of stable collagen triple-helical and coiled coil type structures and does not depend upon the presence of the cytoplasmic or transmembrane domains of this protein. PMID- 9220969 TI - Identification of three key residues in substrate recognition site 5 of human cytochrome P450 3A4 by cassette and site-directed mutagenesis. AB - Cassette mutagenesis and site-directed mutagenesis were used to investigate the importance of individual amino acid residues at positions 364-377 of cytochrome P450 3A4 in determining steroid hydroxylation or stimulation by alpha naphthoflavone. The mutants were expressed in an Escherichia coli system, and solubilized membranes were prepared. All mutants except R365G and R365K exhibited anti-3A immunoreactivity on Western blotting, although R372S and R375K were not detected as the Fe2+-CO complex. Replacement of Arg-372 by Lys yielded a typical P450 spectrum. The results indicate that the highly conserved Arg residues at positions 365 and 375 may play a role in stabilizing the tertiary structure or in heme binding. Catalytic activities of 12 mutants were examined using progesterone and testosterone as substrates, and residues 369, 370, and 373 were found to play an important role in determining substrate specificity. Although the three mutants hydroxylated progesterone and testosterone primarily at the 6beta position like the wild-type, replacement of Ile-369 by Val suppressed progesterone 16alpha-hydroxylase activity, whereas substitution of Ala-370 with Val enhanced progesterone 16alpha-hydroxylation. Interestingly, substitution of Leu-373 with His resulted in production of a new metabolite from both steroids. Moreover, the mutants at positions 369 and 373 were more and less responsive, respectively, than the wild-type to alpha-naphthoflavone stimulation. Alterations in activities or expression of several mutants were interpreted using a three dimensional model of P450 3A4. The results suggest that analogy with mammalian family 2 and bacterial cytochromes P450 can be used to predict P450 3A residues that contribute to regiospecific steroid hydroxylation. PMID- 9220970 TI - Measurement of spontaneous transfer and transbilayer movement of BODIPY-labeled lipids in lipid vesicles. AB - An assay was developed to study the spontaneous transfer and transbilayer movement (flip-flop) of lipid analogs labeled with the fluorescent fatty acid, 5 (5,7-dimethyl BODIPY)-1-pentanoic acid (C5-DMB-) in large unilamellar lipid vesicles comprised of 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC). The assay is based on the concentration-dependent changes in fluorescence intensity that occur when donor vesicles containing a C5-DMB-lipid are mixed with nonfluorescent acceptor vesicles. A kinetic model was developed to describe the time-dependent changes in concentration of a lipid undergoing both spontaneous transfer between unilamellar vesicles and transbilayer movement within the vesicle membranes, and a mathematical solution was obtained. Data were obtained using C5-DMB-labeled analogs of sphingomyelin (C5-DMB-SM), ceramide (C5-DMB-Cer), phosphatidylcholine (C5-DMB-PC), and diacylglycerol (C5-DMB-DAG), and kinetic parameters for each lipid were determined using a nonlinear least-squares fitting program. The half times for interbilayer transfer of the lipids were C5-DMB-SM (21 s) < C5-DMB-PC (350 s) approximately C5-DMB-Cer (400 s) << C5-DMB-DAG (100 h). C5-DMB-Cer (t1/2 approximately 22 min) and C5-DMB-DAG (t1/2 approximately 70 ms) exhibited rapid spontaneous transbilayer movement, while C5-DMB-SM (t1/2 approximately 3.3 h) and C5-DMB-PC (t1/2 approximately 7.5 h) moved across the bilayer very slowly. These results provide a basis for interpreting the behavior of these lipid analogs in cells. PMID- 9220971 TI - Conformational changes in G-CSF/Receptor complex as investigated by isotope edited FTIR spectroscopy. AB - Conformations of G-CSF and the extracellular domain of its receptor as well as their complex have been investigated by employing isotope-edited FTIR spectroscopy. To determine unambiguously the protein conformations of G-CSF and the receptor in the complex, we have prepared uniformly 13C/15N isotope labeled G CSF to resolve its amide I' band from that of the receptor in the IR spectrum of the complex. By comparing the IR spectra of the isotope-labeled G-CSF and the receptor with that of the complex, we have provided spectral evidence that the AB loop region involving the unique 310 helix segment of G-CSF likely undergoes a conformational change to a regular alpha-helix upon binding to the receptor. The IR data also indicate a possible minor increase in alpha-helical conformation for the receptor in the complex. Furthermore, FTIR spectra of G-CSF, the receptor, and their complex demonstrate clearly that protein conformations of both G-CSF and the receptor have been dramatically stabilized by complex formation. Specifically, the melting transition (Tm value) of the alpha-helix in G-CSF is increased by nearly 30 degrees C and that of the beta-strand in the receptor by nearly 15 degrees C in the G-CSF/receptor complex. We estimate from the current FTIR data that the native conformations of approximately 15% of all receptor residues are stabilized by G-CSF binding. On the other hand, the entire alpha helical content of G-CSF appears to be stabilized in the complex. Together, these results indicate that formation of the ligand/receptor complex results in not only conformational changes in the receptor but also significant structural changes in the ligand. This adds insight to the general consensus that binding of ligand to cytokine receptors induces mostly structural changes in the receptor which lead to receptor oligomerization and signal transduction. The current data also suggest a possible physiological role of the 310 helix present in G-CSF for its receptor binding activity. PMID- 9220972 TI - Role of the amino terminus of the third intracellular loop in agonist-promoted downregulation of the alpha2A-adrenergic receptor. AB - A prominent feature of long-term regulation of the alpha2A-adrenergic receptor (alpha2AAR) is a loss of cellular receptors over time (downregulation). The molecular determinants of downregulation were sought by targeting regions of the receptor involved in G protein coupling and phosphorylation. Mutated receptors, consisting of chimeric substitutions of analogous beta2-adrenergic receptor (beta2AR) and serotonin 5-hydroxytryptamine1A (5-HT1A) receptor sequence into the second intracellular loop (ICL2) (residues 113-149), the amino terminus (residues 218-235) and carboxy terminus (residues 355-371) of ICL3, and a deletion of the beta-adrenergic receptor kinase (betaARK) phosphorylation sites in the third intracellular loop (ICL3) (residues 293-304), were expressed in Chinese hamster ovary (CHO) cells. Wild-type alpha2AAR underwent 31% +/- 3% downregulation after 24 h of exposure to 100 microM epinephrine. Loss of downregulation was observed with some mutants, but this was not related to functional coupling to inhibitory or stimulatory guanine nucleotide regulatory binding proteins (Gi or GS) or to phosphorylation. Rather, any mutant with a substitution of the amino terminus of ICL3 (regardless of whether the substitution was with beta2AR or 5-HT1A sequence) resulted in upregulation. Studies with an inhibitor of protein synthesis indicated that the primary mechanism of downregulation of the alpha2AAR is agonist-promoted degradation of receptor protein which requires a destabilization sequence in the amino terminus of ICL3. Thus, in contrast to other G protein coupled receptors, in which G protein coupling or phosphorylation are critical for long-term agonist regulation, the alpha2AAR has a specific structural domain distinct from these other functional regions that serves to direct agonist promoted downregulation. PMID- 9220973 TI - Entropy-driven interactions of anesthetics with membrane proteins. AB - Thermodynamic analysis of anesthetic effects on Ca2+-ATPase activity was performed to evaluate the feasibility of anesthetic binding and gain insight into the molecular events underlying the anesthetic-enzyme interactions. The Ca2+ ATPases, integral membrane proteins vital in cellular Ca2+ regulation, are suitable models for investigation of the mechanism of anesthetic action on membrane proteins that are targeted by the anesthetics. Ca2+-ATPase of plasma membrane, PMCA, and SERCA1 in the intracellular sarcoplasmic reticulum membrane were used to study two general anesthetics: halothane, a halogenated two-carbon alkane; and propofol, an intravenous, strongly lipophilic-substituted phenol. Interactions of both anesthetics result in a negative Gibbs free energy change, which in both enzymes is more favorable for the more lipophilic propofol than halothane. Temperature dependence (more negative change in Gibbs free energy at increased temperature) is in agreement with predominantly nonpolar interactions. The interactions are entropy-driven, characterized by positive enthalpy which is overcompensated by positive entropy changes. This is in contrast to the reported in literature enthalpy-driven anesthetic binding to soluble proteins. The possible contributions to the observed positive entropy change are discussed including displacement of ordered water molecules by anesthetic binding in nonpolar cavities in the membrane proteins and subtle structural rearrangements. PMID- 9220974 TI - Evidence for multiple mechanisms for membrane binding and integration via carboxyl-terminal insertion sequences. AB - Subcellular localization of proteins with carboxyl-terminal insertion sequences requires the molecule be both targeted to and integrated into the correct membrane. The mechanism of membrane integration of cytochrome b5 has been shown to be promiscuous, spontaneous, nonsaturable, and independent of membrane proteins. Thus endoplasmic reticulum localization for cytochrome b5 depends primarily on accurate targeting to the appropriate membrane. Here direct comparison of this mechanism with that of three other proteins integrated into membranes via carboxyl-terminal insertion sequences [vesicle-associated membrane protein 1(Vamp1), polyomavirus middle-T antigen, and Bcl-2] revealed that, unlike cytochrome b5, membrane selectivity for these molecules is conferred at least in part by the mechanisms of membrane integration. Bcl-2 membrane integration was similar to that of cytochrome b5 except that insertion into lipid vesicles was inefficient. Unlike cytochrome b5 and Bcl-2, Vamp1 binding to canine pancreatic microsomes was saturable, ATP-dependent, and abolished by mild trypsin treatment of microsomes. Surprisingly, although the insertion sequence of polyomavirus middle-T antigen was sufficient to mediate electrostatic binding to membranes, binding did not lead to integration into the bilayer. Together these results demonstrate that there are at least two different mechanisms for correct membrane integration of proteins with insertion sequences, one mediated primarily by targeting and one relying on factors in the target membrane to mediate selective integration. Our results also demonstrate that, contrary to expectation, hydrophobicity is not sufficient for insertion sequence-mediated membrane integration. We suggest that the structure of the insertion sequence determines whether or not specific membrane-bound receptor proteins are required for membrane integration. PMID- 9220975 TI - Alteration of substrate specificity by mutations at the His61 position in predicted transmembrane domain 1 of human MDR1/P-glycoprotein. AB - In CFTR, a member of the ABC superfamily and a chloride channel, amino acid substitutions in its transmembrane domains 1 and 6 (TM1, TM6) have been reported to modulate the anion selectivity or ion conductance of the ion channel. In P glycoprotein, no amino acid substitution in TM1, but some in TM6, have been reported to modify the substrate specificity of this protein. In this work, we demonstrated the involvement of His61, which is in the middle of the predicted TM1, in the function of P-glycoprotein. His61 was replaced by all other amino acid residues, and each of the mutant cDNAs was introduced into drug-sensitive human carcinoma cells, KB3-1. The drug-resistance profile of cells stably expressing each mutated P-glycoprotein was investigated by comparing their relative resistance to vinblastine, colchicine, VP16, and adriamycin. The resistance to vinblastine was increased by replacing His61 by amino acids with smaller side chains, while it was lowered by replacing by amino acids with bulkier side chains. The reverse effect was observed for resistance to colchicine and VP16. The resistance to adriamycin was increased by replacing by amino acids with bulkier side chains except Lys or Arg, which have a basic side chain. We also showed that the replacement of His61 by Phe and Lys greatly impaired the efflux of calcein AM, while the replacement had no effect on the efflux of rhodamine 123. These results suggest that an amino acid residue at position 61 in TM1 is important in deciding the substrate specificity of P-glycoprotein. PMID- 9220976 TI - Conformational changes and fusion activity of vesicular stomatitis virus glycoprotein: [125I]iodonaphthyl azide photolabeling studies in biological membranes. AB - The interaction of VSV glycoprotein (VSV G) with biological membranes was studied by photosensitized labeling. The method is based on photosensitized activation by the fluorescent lipid analog 3,3'-dioctadecyloxacarbocyanine (DiO) of a hydrophobic probe, [125I]iodonaphthyl azide (125INA), that rapidly partitions into the membrane bilayer of virus and cells. 125INA labeling of proteins and lipids can be confined to the site of chromophore localization by photosensitized labeling. Photoactivation using visible light of target membrane labeled with DiO and 125INA, to which unlabeled virions are bound, results in exclusive labeling of envelope glycoproteins inserted into the target membrane [Pak et al. (1994) J. Biol. Chem. 269, 14614]. In this study, we labeled lipid symmetric erythrocyte ghosts with 125INA and DiO. Photosensitized activation of VSV prebound to labeled ghosts with visible light resulted in VSV G labeling under fusogenic conditions. Photoactivation of 125INA by UV light, which is nonspecific, produced labeled VSV G at both acidic and neutral pH. Photosensitized labeling of VSV G by DiO-125INA ghosts was also observed at pH 5.5, 4 degrees C, in the absence of mixing between viral and cellular lipids, suggesting insertion of the ectodomain of VSV G. Soluble VSV G lacking the transmembrane domain inserted into DiO-125INA-ghosts under the same conditions as intact VSV G. DiO inserted into intact VSV appeared to be a suitable fluorophore for continuous kinetic measurements of membrane fusion by fluorescence dequenching. Our photosensitized labeling results establish biochemical correlates for the three states of VSV G, which we had proposed based on kinetic data [Clague et al., Biochemistry 29, 1303]. In addition, we found that VSV G insertion into the target membrane is reversible, suggesting a "velcro"-like attachment of the fusogenic domain with the target membrane. PMID- 9220977 TI - Structure and thermal stability of photosystem II reaction centers studied by infrared spectroscopy. AB - The secondary structure of photosystem II reaction centers isolated from pea has been deduced from quantitative analysis of the component bands of the infrared amide I spectral region, determined by FTIR spectroscopy. The analysis shows the isolated complex to consist of 40% alpha-helix, 10% beta-sheet, 14% beta-strands (or extended chains), 17% turns, 15% loops, and 3% nonordered segments. These structural protein elements were determined for samples in H2O, in D2O, and in dried films. The isolated reaction center, composed of proteins D1,D2,cytochrome b559, and PsbI, has been predicted to contain a total of 13 transmembrane alpha helices, which conveys a percentage of this type of structure congruent with the structural determination deduced from FTIR spectra. The process of thermal destabilization of the reaction centers has also been studied by FTIR spectroscopy, showing a clear main conformational transition at 42 degrees C, which indicates a high thermal sensitivity of the secondary structure of this protein complex. Such thermal instability may correlate with the well-described high sensitivity of photosystem II to damage and may relate to the process of rapid protein degradation that photosystem II suffers during photoinhibition of plants. PMID- 9220978 TI - Analysis of the reaction coordinate of photosynthetic water oxidation by kinetic measurements of 355 nm absorption changes at different temperatures in photosystem II preparations suspended in either H2O or D2O. AB - Flash-induced absorption changes at 355 nm were measured at different temperatures within the range of 2 degrees C S2) = 14 kJ/mol, EA(S2-->S3) = 35 kJ/mol, and EA(S3-->-->S0 + O2) = 21 kJ/mol for theta > 11 degrees C, 67 kJ/mol for theta < 11 degrees C in PS II core complexes dissolved in H2O; (b) replacement of exchangeable protons by deuterons causes only minor changes ( S2, S2 --> S3, and S3 -->--> S0 + O2, respectively. The corresponding values of PS II membrane fragments are 1.3, 1.3, and 1. 4. Based on these results and corresponding EA data reported in the literature for PS II membrane fragments from spinach [Renger, G., & Hanssum, B. (1992) FEBS Lett. 299, 28-32] and PS II particles from the thermophilic cyanobacterium Synechococcus vulcanus Copeland [Koike, H., Hanssum, B., Inoue, Y., & Renger, G. (1987) Biochim. Biophys. Acta 893, 524-533], the reaction coordinate of the redox sequence in the WOC is inferred to be almost invariant to the evolutionary development from cyanobacteria to higher plants. Furthermore, the rather high activation energy of the S2 --> S3 transition provides evidence for a significant structural change coupled with this reaction. Implications for the mechanism of photosynthetic water oxidation are discussed. PMID- 9220979 TI - Quantitative kinetic model for photoassembly of the photosynthetic water oxidase from its inorganic constituents: requirements for manganese and calcium in the kinetically resolved steps,. AB - The process of photoactivation, the assembly of a functional water-oxidizing complex (WOC) from the apoproteins of photosystem II of higher plants and inorganic cofactors (Mn2+, Ca2+, and Cl-), was known from earlier works to be a two-step kinetic process, requiring two light-induced processes separated by a slower dark period. However, these steps had not been directly resolved in any kinetic experiment, until development of an ultrasensitive polarographic O2 electrode and synthesis of an improved chelator for cofactor removal allowed direct kinetic resolution of the first pre-steady state intermediate [Ananyev, G. M. & Dismukes, G. C. (1996a) Biochemistry 35, 4102-4109]. Herein, the dependence of the rates of each of the first two light steps and the dark step of photoactivation was directly determined in spinach PSII membranes over a range of calcium and manganese concentrations at least 10-fold lower than those possible using commercial O2 electrodes. The following results were obtained. (1) One Mn2+ ion binds and is photooxidized to Mn3+ at a high-affinity site, forming the first light-induced intermediate, IM1. Formation of IM1 is coupled to the dissociation of a bound Ca2+ ion either located in the Mn site or coupled to it. (2) The inhibition constant for Ca2+ dissociation from this site is equal to 1.5 mM. (3) The dissociation constant of Mn2+ at this high-affinity site is equal to 8 microM at the optimum calcium concentration for O2-evolving activity of 8 mM, in agreement with the high-affinity site for electron donation to PSII. (4) Prior to the next photolytic step, one Ca2+ ion must bind at its effector site so that stable photooxidation of a second Mn2+ ion can occur, forming the second light induced intermediate, IM2. This dark process is the rate-determining step. (5) The Michaelis constant for recovery of O2 evolution by Ca2+ binding at this effector site (Km) is equal to 1.4 mM, a value that is the same as that measured for the calcium requirement for O2 evolution in intact PSII. (6) The low quantum yield for the formation of IM2 from IM1 increases linearly with the duration of the dark period up to the longest period we could examine (10 s). Accordingly, the rate limitation in the second photolytic step originates from a slow calcium induced dark rearrangement of the first intermediate, IM1, which we propose to be a protein conformational change that allows stable binding of the next Mn2+ ion. We further propose that the single Ca2+ ion which is required for assembly of the Mn4 cluster is equivalent to the Ca2+ ion which functions at the "gatekeeper" site in intact O2-evolving centers, where it plays a role in limiting substrate access to the Mn4 cluster [Sivaraja, M., et al. (1989) Biochemistry 28, 9459 9464; Tso, J., et al., (1991) Biochemistry 30, 4734-4739]. A molecular model for photoactivation is proposed and discussed. PMID- 9220980 TI - Mutagenesis of a proton linkage pathway in Escherichia coli manganese superoxide dismutase. AB - Mutagenesis of Escherichia coli manganese superoxide dismutase (MnSD) demonstrates involvement of the strictly conserved gateway tyrosine (Y34) in exogenous ligand interactions. Conservative replacement of this residue by phenylalanine (Y34F) affects the pH sensitivity of the active-site metal ion and perturbs ligand binding, stabilizing a temperature-independent six-coordinate azide complex. Mutant complexes characterized by optical and electron paramagnetic resonance (EPR) spectroscopy are distinct from the corresponding wild-type forms and the anion affinities are altered, consistent with modified basicity of the metal ligands. However, dismutase activity is only slightly reduced by mutagenesis, implying that tyrosine-34 is not essential for catalysis and may function indirectly as a proton donor for turnover, coupled to a protonation cycle of the metal ligands. In vivo substitution of Fe for Mn in the MnSD wild-type and mutant proteins leads to increased affinity for azide and altered active-site properties, shifting the pH-dependent transition of the active site from 9.7 (Mn) to 6.4 (Fe) for wt enzyme. This pH-coupled transition shifts once more to a higher effective pKa for Y34F Fe2-MnSD, allowing the mutant to be catalytically active well into the physiological pH range and decreasing the metal selectivity of the enzyme. Peroxide sensitivities of the Fe complexes are distinct for the wild-type and mutant proteins, indicating a role for Y34 in peroxide interactions. These results provide evidence for a conserved peroxide protonation linkage pathway in superoxide dismutases, analogous to the proton relay chains of peroxidases, and suggests that the selectivity of Mn and Fe superoxide dismutases is determined by proton coupling with metal ligands. PMID- 9220981 TI - Resonance Raman study on the oxidized and anionic semiquinone forms of flavocytochrome b2 and L-lactate monooxygenase. Influence of the structure and environment of the isoalloxazine ring on the flavin function. AB - The oxidized and semiquinone anion radical forms of flavin mononucleotide carried by flavocytochrome b2 and L-lactate monooxygenase have been studied by resonance Raman (RR) spectroscopy. The RR spectra of their oxidized forms are compared with previously published RR data on various flavins and flavoproteins. Taking as a support available X-ray crystallographic data on flavoproteins, we have found correlations between the frequencies of RR bands II (1575-1588 cm-1), III (1534 1557 cm-1), and X (1244-1266 cm-1) and the H-bonding environment and/or the structure of the flavin ring. The present RR data provide strong evidence that the electron density, the conformation, and the H-bonding environment of the oxidized flavin mononucleotide of flavocytochrome b2 and L-lactate monooxygenase are different. As far as the anionic semiquinone form of flavoproteins is concerned, the behavior of two bands observed at 1280-1300 and 1320-1350 cm-1 suggests that they have vibrational origins similar to those of RR bands II and III of oxidized compounds. On this basis, the differences in conformation and H bonding environment of the isoalloxazine ring, observed for the oxidized form of flavocytochrome b2 and L-lactate monooxygenase, appear to be preserved upon one electron reduction of the flavin. For both flavoproteins, the RR spectra of the semiquinone form are affected by pyruvate binding. The data are interpreted in the frame of a change in H-bonding interaction of the C4&dbd;O carbonyl group of the flavin without significant alteration of the isoalloxazine conformation. This modification in electrostatic interaction quantitatively accounts for the pyruvate-induced changes of the oxidized/semiquinone and semiquinone/reduced redox potentials of the flavoproteins. Considering the high homology in the flavin catalytic sites of flavocytochrome b2 and L-lactate monooxygenase, the observed differences in H-bonding environment and conformation of the FMN ring are related to the different biological functions of the two flavoproteins. PMID- 9220982 TI - Fluoride binding in hemoproteins: the importance of the distal cavity structure. AB - The electronic absorption and resonance Raman spectra of the fluoride complexes of various peroxidases and selected site-directed mutants have been studied at pH 5.0, and compared to the spectra obtained for the myoglobin-F adduct. It is shown that the electronic absorption maxima depend on the degree of conjugation between the porphyrin macrocycle and the vinyl substituents. Moreover, it is confirmed that the wavelength of the CT1 band is a sensitive probe of axial ligand polarity and of its interaction with the distal protein residues. The results highlight the different mechanism of stabilization of the fluoride ligand exerted by the distal residues in myoglobin and peroxidases. In peroxidases, the Arg is determinant in controlling the ligand binding via a strong hydrogen bond between the positively charged guanidinium group and the anion. Mutation of Arg to Leu decreases the stability of the complex by 900-fold, suggesting that this interaction stabilizes the complex by 4 kcal/mol. The distal His also contributes to the stability of the fluoride complex, presumably by accepting a proton from HF and hydrogen-bonding, through a water molecule, to the anion. Mutation of His to Leu decreases the stability of the fluoride complex by 30-fold, suggesting that this interaction is much weaker than the interaction with the distal Arg. For Mb, the distal His is solely responsible for stabilization of the exogenous ligand. PMID- 9220984 TI - Mutational analysis of an antigenic peptide shows recognition in a loop conformation. AB - We have analyzed the recognition between an antigenic undecapeptide and a monoclonal antibody through a mutational approach. Antibody mAb164 is directed against the native form of the TrpB2 subunit of Escherichia coli tryptophan synthase. It recognizes a synthetic peptide, P11, constituted of residues 273 HGRVGIYFGMK-283 of TrpB with high affinity. P11 was fused with a carrier protein, MalE, to facilitate its manipulation. The affinities between mAb164 and the MalE P11 hybrids were measured by competition enzyme-linked immunosorbent assay (ELISA). The changes of the P11 residues into progressively shorter residues, the comparison of changes into Pro and Ala, and the study of double mutants showed the following. Four hydrophobic residues of P11, Val276, Ile278, Tyr279, and Phe280, were predominant in the interaction. For some residues, e.g., Tyr279, most groups of the side chain contributed to the interaction. For others, only some groups played a significant role, e.g., the Cdelta group of Ile278 or the Cbeta group of Phe280. The lack of side chain in position Gly281 and a tertiary interaction between the side chains of Ile278 and Lys283 were important. P11 was recognized in a loop conformation, close to that of residues 273-283 of TrpB in the crystal structure of the complete tryptophan synthase, TrpA2TrpB2. Comparison of our mutational data with NMR data on the conformation of the isolated peptide P11 and with kinetic data on its interaction with mAb164 indicate that mAb164 selects a conformer of P11 that represents only a small minority of the molecules. Our results provide useful information on the mechanisms by which linear epitopes and unconstrained peptides are recognized by receptors. PMID- 9220983 TI - Conformational and functional properties of an undecapeptide epitope fused with the C-terminal end of the maltose binding protein. AB - Monoclonal antibody mAb164 is directed against the TrpB2 subunit of the Escherichia coli tryptophan synthase. It recognizes the synthetic peptide P11, constituted of residues 273-283 of TrpB, with high affinity. We constructed a hybrid protein in which the C-terminal end of protein MalE was linked with the N terminal end of P11. Hybrid MalE-P11 was produced in E. coli from a plasmidic gene and purified in one step as MalE. MalE-P11 and the isolated P11 had identical conformational and functional properties according to the following criteria. The NMR spectra of MalE and MalE-P11 in TOCSY experiments showed that the P11 moiety of MalE-P11 moved independently from its MalE moiety. The chemical shifts of the protons for the P11 moiety of MalE-P11 and for the isolated P11 were very close and did not show significant deviations from random coil values. The equilibrium constant of dissociation (KD) from mAb164, measured by a competition ELISA, was identical for MalE-P11 and the isolated P11, around 6 nM. The change of the C-terminal residue of MalE-P11 from Lys into Ala increased 37 fold this dissociation constant. This increase showed that the P11 moiety of MalE P11 was not degraded. The high molecular mass of MalE-P11 allowed us to follow its kinetics of interaction with immobilized mAb164 by surface plasmon resonance, using the BIAcore apparatus. The rates of association with mAb164 were similar for MalE-P11 and TrpB2, but the dissociation was faster for MalE-P11 than for TrpB2, as previously observed for the isolated P11 by a fluorometric method. Thus, the fusion of peptides with the C-terminal end of MalE could constitute an alternative to chemical synthesis for the study of their recognition by receptors, in vivo or in vitro. PMID- 9220985 TI - Influence of Arginines 93, 97, and 101 of thrombin to its functional specificity. AB - Mutation of three Arg residues, 93, 97, and 101, to Ala in thrombin (thrombin R93,97,101A) has previously been shown to eliminate most heparin acceleration of thrombin inhibition by antithrombin and most of the ability of chondroitin sulfate (CS) on thrombomodulin (TM) to enhance affinity for TM and to eliminate the characteristic high-affinity interaction with protein C observed with TM lacking CS. In this study we examined the relative impact of mutation of these Arg residues alone and in combination on the above reactions and, in addition, on the ability of rabbit TM to accelerate thrombin inhibition by antithrombin. The order of importance for heparin acceleration of inhibition by antithrombin was Arg 101, 93, and 97. In contrast, Arg 97 was the major residue required for TM dependent acceleration of reactivity with antithrombin and for CS-dependent enhancement of TM affinity. Arg 101 and 93 were critical for TM-dependent, high affinity protein C interaction at low Ca2+ concentrations, while Arg 97, which was critical for the other TM-dependent effects, played no detectable role in this metal dependence. These results illustrate that these Arg residues in anion binding exosite 2 contribute very differently to the diverse reactions dependent on that domain in thrombin. PMID- 9220987 TI - Solution structure of oxidized Saccharomyces cerevisiae iso-1-cytochrome c. AB - The solution structure of oxidized Saccharomycescerevisiae Cys102Ser iso-1 cytochromechas been determined using 1361 meaningful NOEs (of 1676 total) after extending the published proton assignment [Gao, Y., et al. (1990) Biochemistry 29, 6994-7003] to 77% of all proton resonances. The NOE patterns indicate that secondary structure elements are maintained upon oxidation in solution with respect to the solid state and solution structures of the reduced species. Constraints derived from the pseudocontact shifts [diamagnetic reference shift values are those of the reduced protein [Baistrocchi, P., et al. (1996) Biochemistry 35, 13788-13796]] were used in the final stages of structure calculations. After restrained energy minimization with constraints from NOEs and pseudocontact shifts, a family of 20 structures with rmsd values of 0.58 +/- 0.08 and 1.05 +/- 0.10 A (relative to the average structure) for the backbone and all heavy atoms, respectively, was obtained. The solution structure is compared with the crystal structure and the structures of related systems. Twenty-six amide protons were detected in the NMR spectrum 6 days after the oxidized lyophilized protein was dissolved in D2O (pH 7.0 and 303 K); in an analogous experiment, 47 protons were observed in the spectrum of the reduced protein. The decrease in the number of nonexchanging amide protons, which mainly are found in the loop regions 14-26 and 75-82, confirms the greater flexibility of the structure of oxidized cytochrome c in solution. Our finding of increased solvent accessibility in these loop regions is consistent with proposals that an early step in unfolding the oxidized protein is the opening of the 70-85 loop coupled with dissociation of the Met80-iron bond. PMID- 9220986 TI - Structural and dynamical properties of a denatured protein. Heteronuclear 3D NMR experiments and theoretical simulations of lysozyme in 8 M urea. AB - Oxidized and reduced hen lysozyme denatured in 8 M urea at low pH have been studied in detail by NMR methods. 15N correlated NOESY and TOCSY experiments have provided near complete sequential assignment for both 1H and 15N resonances. Over 900 NOEs, including 130 (i, i + 2) and 23 (i, i + 3) NOEs, could be identified by analysis of the NOESY spectra of the denatured states, and 3J(HN, Halpha) coupling constants and 15N relaxation rates have been measured. The coupling constant and NOE data were analyzed by comparisons with theoretical predictions from a random coil polypeptide model based on amino acid specific phi,psi distributions extracted from the protein data bank. There is significant agreement between predicted and experimental NMR parameters suggesting that local conformations of the denatured states are largely determined by short-range interactions within the polypeptide chain. This result is supported by the observation that the chemical shift, coupling constant, and NOE data are little affected by whether or not the four disulfide bridge cross-links are formed in the denatured protein. The relaxation data, however, show significant differences between the oxidized and reduced protein. Analysis of the relaxation data in terms of simple dynamics models provides evidence for weak clustering of hydrophobic groups near tryptophan residues and increased barriers to motion in the more compact conformers formed when the polypeptide chain is cross-linked by the disulfide bridges. Using this information, a structural description of these denatured states is given in terms of an ensemble of conformers, which have a complex relationship between their local and global characteristics. PMID- 9220988 TI - Crystal structure of Kex1deltap, a prohormone-processing carboxypeptidase from Saccharomyces cerevisiae,. AB - Kex1p is a prohormone-processing serine carboxypeptidase found in Saccharomyces cerevisiae. In contrast to yeast serine carboxypeptidase (CPD-Y) and wheat serine carboxypeptidase II (CPDW-II), Kex1p displays a very narrow specificity for lysyl or arginyl residues at the C-terminus of the substrate. The structure of Kex1Deltap, an enzyme that lacks the acidic domain and membrane-spanning portion of Kex1p, has been solved by a combination of molecular replacement and multiple isomorphous replacement and refined to a resolution of 2.4 A. The S1' site of Kex1Deltap is sterically restricted compared to those from CPD-Y or CPDW-II; it also contains two acidic groups that are well positioned to interact with the basic group of a lysine or arginine side chain. The high specificity of Kex1p can therefore be explained by a combination of steric and electronic factors. The structure of the S1 site of Kex1Deltap is also well suited for binding of a lysine or arginine side chain, and the enzyme may therefore exhibit a preference for these residues at P1. PMID- 9220989 TI - Regulation of oxidation-reduction potentials through redox-linked ionization in the Y98H mutant of the Desulfovibrio vulgaris [Hildenborough] flavodoxin: direct proton nuclear magnetic resonance spectroscopic evidence for the redox-dependent shift in the pKa of Histidine-98. AB - Flavodoxin from Desulfovibrio vulgaris is a low molecular weight (15 000 Da) acidic flavoprotein that contains a single flavin mononucleotide (FMN) cofactor. A distinguishing feature of the flavodoxin family is the exceptionally low midpoint potential of the semiquinone/hydroquinone couple. Tyrosine-98, which flanks the outer or si face of the FMN, plays an important role in establishing the oxidation-reduction properties of the bound cofactor as demonstrated by the substitution of a number of amino acids at this position [Swenson, R. P., & Krey, G. D. (1994) Biochemistry 33, 8505-8514]. The midpoint potential for the semiquinone/hydroquinone couple increases substantially when basic residues are introduced at this position. The pH dependency in the Y98H mutant is consistent with a redox-linked ionization model in which the favorable electrostatic coupling between the imidazolium cation and the flavin hydroquinone anion is responsible for the higher potential. Such a model predicts an increase in the pKa of 1.5 units for His98 upon complete reduction of the FMN. In this study, proton nuclear magnetic resonance spectroscopy was used to directly determine the intrinsic pKa of His98 as a function of the redox state of the cofactor in this flavodoxin. Values for the pKa of His98 in the oxidized and fully reduced flavodoxin are 7.02 +/- 0.08 and 8.43 +/- 0.11, respectively, an increase in the pKa by 1.41 units, which conforms with the previous prediction. These results provide direct experimental proof of the redox-linked ionization of this residue and provides further evidence of the crucial role of electrostatic interactions, in this case, in the stabilization of the flavin hydroquinone anion. This phenomenon may represent a general mechanism in the modulation of the reduction potential of the flavin cofactor within flavoenzymes in which ionizable groups such as histidine in the active center change ionization states during the catalytic cycle. PMID- 9220990 TI - Mechanism-based inhibitors for the inactivation of the bacterial phosphotriesterase. AB - 1-Bromovinyl (I), Z-2-bromovinyl (II), 1,2-dibromoethyl (III), and a series of 4 (halomethyl)-2-nitrophenyl (IVa-c) diethyl phosphate esters were examined as substrates and mechanism-based inhibitors for the bacterial phosphotriesterase. All of these compounds were found to act as substrates for the enzyme. Inhibitor I rapidly inactivated the enzyme within 1 min, giving a partition ratio of 230. The newly formed covalent adduct with inhibitor I was susceptible to hydrolysis at elevated values of pH and dissociation by NH2OH. Azide was not able to protect the enzyme from inactivation with inhibitor I, implying that the reactive species was not released into solution prior to the inactivation event. The reactive species was proposed to be either an acyl bromide or a ketene intermediate formed by the enzymatic hydrolysis of inhibitor I. Compounds II and III were shown to be relatively poor substrates of phosphotriesterase and they did not induce any significant inactivation of the enzyme. The inhibitor, 4-(bromomethyl)-2 nitrophenyl diethyl phosphate (IVa), was found to irreversibly inactivate the enzyme with a KI = 7.9 mM and kinact = 1. 2 min-1 at pH 9.0. There was no effect on the rate of inactivation upon the addition of the exogenous nucleophiles, azide, and NH2OH. The species responsible for the covalent modification of the enzyme by IVa was most likely a quinone methide formed by the elimination of bromide from the phenolic intermediate. NMR experiments demonstrated that the quinone methide did not accumulate in solution. The chloro (IVb) and fluoro (IVc) analogues did not inactivate the enzyme. These results suggest that the elimination of the halide ion from the phenolic intermediate largely determines the partition ratio for inactivation. PMID- 9220991 TI - Affinity radiolabeling identifies peptides associated with the isomerase activity of human type I (placental) 3beta-hydroxysteroid dehydrogenase/isomerase. AB - 3beta-Hydroxysteroid dehydrogenase and steroid Delta5-->4-isomerase (3beta HSD/isomerase) were purified as a single protein from human term placenta. The affinity alkylator, 5,10-secoestr-4-yne-3,10, 17-trione (secosteroid), was incubated with the purified enzyme (30/1 secosteroid/enzyme molar ratio) to produce an 80% loss of initial isomerase activity over 90 min in a time dependent, irreversible manner. The secosteroid inactivated 3beta-HSD by only 20% during the same 90 min. Incubations containing the isomerase substrate steroid, 5 androstene-3,17-dione, completely protected the isomerase activity from inactivation by the secosteroid and did not slow the inactivation of 3beta-HSD. The enzyme containing covalently bound steroid was separated from unreacted secosteroid by reversed phase HPLC. Ketones on the protein-bound secosteroid were radiolabeled by reduction with sodium boro[3H]hydride (specific radioactivity 50 microCi/micromol for the transferred tritium). After removal of the unreacted sodium boro[3H]hydride, the affinity-radiolabeled enzyme was digested with trypsin-TPCK, and the peptides were isolated by reversed phase HPLC. The radiolabeled peptide fractions were sequenced. The secosteroid alkylated three tryptic peptides: 251GQFYYISDDTPHQSYDNLNYTLSK274, tritiated His262; 176NGGTLYTCALR186, tritiated Cys183; and 353TVEWVGSLVDR363, tritiated Trp356. Coincubation with the isomerase substrate blocked the labeling of these three peptides and shifted the alkylation by secosteroid to a single tryptic peptide (135EIIQNGHEEEPLENTWPAPYPHSK159, tritiated His142). Using substrate protection to validate specificity, the affinity labeling secosteroid has identified peptides in the enzyme that are associated with isomerase activity. PMID- 9220992 TI - Caged NADP and NAD. Synthesis and characterization of functionally distinct caged compounds. AB - Two caged NADP compounds have been synthesized and characterized for use in the crystallographic study of isocitrate dehydrogenase (IDH), as well as for general use in cell biology, metabolism, and enzymology. One caged NADP compound has been designed to be "catalytically caged" so that it can bind to IDH prior to photolysis but is not catalytically active. A second NADP compound is "affinity caged" so that addition of the caging group inhibits binding of the compound to IDH prior to photolysis. The catalytically caged compound was synthesized in a two-step process, starting with the NADase-catalyzed exchange of a synthetic nicotinamide derivative onto NADP. X-ray structures of the NADP compounds with IDH show the catalytically caged NADP bound to the enzyme with its nicotinamide group improperly positioned to allow turnover, while the affinity caged NADP does not bind to the enzyme at concentrations up to 50 mM. Two analogous caged NAD compounds have also been synthesized. The NADP and NAD compounds were characterized in terms of kinetics, quantum yield, and product formation. The affinity caged NADP compound P2'-[1-(4,5-dimethoxy-2-nitrophenyl)ethyl] NADP (VIII) is photolyzed at a rate of 1.8 x 10(4) s-1 with a quantum yield of 0.19 at pH 7; the NAD analog P-[1-(4,5-dimethoxy-2-nitrophenyl)ethyl] NAD (IX) is photolyzed at at a rate of 1.7 x 10(4) s-1 with a quantum yield of 0.17. PMID- 9220994 TI - The 9-arginine residue of alpha-conotoxin GI is responsible for its selective high affinity for the alphagamma agonist site on the electric organ acetylcholine receptor. AB - The two agonist-binding domains of the electric organ nicotinic acetylcholine receptor are located at the alphagamma and alphadelta subunit interfaces. alpha Conotoxins GI and MI are competitive antagonists of this receptor and, like d tubocurarine, bind to the alphagamma site with much higher affinity than to the alphadelta site. In the present study, alpha-conotoxin SIA also displayed strong affinity for the alphagamma site but no measurable affinity for the alphadelta site, thus showing even greater site-selectivity. In contrast, alpha-conotoxin SI does not distinguish between the two agonist sites, although its sequence differs from that of GI at only three positions: GI, ECCNPACGRHYSC; SI, ICCNPACGPKYSC. Analogues of SI and GI modified at these three positions were studied to identify the determinants of GI's alphagamma selectivity. Substituting arginine for proline at position 9 produced peptides which displayed "GI-like" selectivity for the alphagamma site. Conversely, substituting proline for arginine at position 9 resulted in "SI-like" nonselective inhibitors. An SI analogue having alanine in place of proline 9 did not distinguish between the two agonist sites and displayed about the same affinity as SI, indicating the importance of the arginyl cation. Interchanging the residues at position 1 or at position 10 influenced the affinity for the receptor but did not measurably change peptide selectivity. Therefore, of the three sequence differences in SI and GI, the variation at position 9, proline and arginine, respectively, is sufficient to account for GI's selective high-affinity binding to the alphagamma site on the electric organ acetylcholine receptor. PMID- 9220995 TI - Effect of preformed correct tertiary interactions on rapid two-state tendamistat folding: evidence for hairpins as initiation sites for beta-sheet formation. AB - The role of preformed correct side chain interactions, such as disulfide bonds, on protein folding kinetics is still not well understood. We investigated the effect of disulfide bond replacements on folding and stability of the small beta sheet protein tendamistat. Tendamistat folds very fast (tau = 10 ms at pH 7 in water) and without detectable intermediates, which facilitates molecular interpretation of the kinetic data. Tendamistat contains two disulfide bonds, one between cysteines 11 and 27, which connects the ends of a beta-hairpin, and a second one between cysteines 45 and 73, which brings together the two outer strands of a three-stranded beta-sheet. Two single-disulfide variants of the protein were prepared by site-directed mutagenesis (tendamistat C11A/C27S and tendamistat C45A/C73A), and the effects on stability and on folding were monitored. Replacement of either disulfide bond leads to a large decrease in protein stability (DeltaDeltaG0 = 6.0 kcal/mol for the C11A/C27S variant and 5.1 kcal/mol for the C45A/C73A variant). This effect is caused both by entropic stabilization of the unfolded state and by enthalpic destabilization of the native structure. Kinetic experiments show that the main effect of fixed side chain contacts is on the unfolding rate. For both single-disulfide variants, unfolding is strongly accelerated (4250 times in the C11A/C27S variant and 250 times in the C45A/C73A variant) whereas the refolding rate constants are only slightly decreased. The activation parameters show that the observed small effect on the refolding reaction in the C11A/C27S variant is a consequence of large and compensating changes in the entropy and enthalpy of activation. Structural interpretation of the kinetic data suggests that formation of the beta-hairpin stabilized by the C11-C27 disulfide bond forms in the rate-limiting step of the refolding process. The interactions between the outer strands of the beta-sheet connected by the C45-C73 disulfide bond, in contrast, are made late in refolding. These results support the idea that beta-hairpins are initiation sites for beta sheet formation and that additional strands are added late in the folding process. PMID- 9220993 TI - Annexin XII forms calcium-dependent multimers in solution and on phospholipid bilayers: a chemical cross-linking study. AB - The annexins are a family of proteins that bind in a Ca2+-dependent manner to phospholipids that are preferentially located on the intracellular face of plasma membranes. Recent X-ray studies of hydra annexin XII showed that it crystallized as a homohexamer with an intermolecular Ca2+ binding site separate from the type II Ca2+-dependent phospholipid binding site. On the basis of this hexamer structure, a novel mechanism was proposed to explain how annexins interact with membranes. The first step toward evaluating this proposal is to determine whether the annexin XII hexamer exists when the protein is not in a crystalline form. We now report that annexin XII in solution can be cross-linked with dimethyl suberimidate into multimers with apparent Mr's corresponding to trimers and hexamers as determined by SDS--polyacrylamide gel electrophoresis--the trimer band may correspond to incompletely cross-linked hexamers. Multimer formation was dependent on Ca2+ and was enhanced when the protein first was bound to phospholipid vesicles. To evaluate the role of the intermolecular Ca2+ site in annexin XII hexamer formation, one of the residues used to coordinate Ca2+, glutamate 105, was replaced with lysine (E105K). In solution, the E105K mutation inhibited hexamer formation in the presence of moderate (3 mM) but not high (25 mM) Ca2+. No inhibition of E105K annexin XII hexamer formation was observed in the presence of phospholipid, thereby suggesting that either (i) other interactions are capable of stabilizing the hexamer when bound to bilayers or (ii) only trimers form on bilayers and the observed hexamer bands were due to cross-linking of closely packed trimers. In summary, this study shows for the first time that annexin XII can form hexamers in solution and implicates the intermolecular Ca2+ site in hexamer formation. This study also shows that multimers form on bilayers but does not clearly establish whether the multimers are trimers or hexamers. PMID- 9220996 TI - Ah receptor nuclear translocator protein heterogeneity is altered after heterodimerization with the Ah receptor. AB - The Ah receptor (AhR) and the Ah receptor nuclear translocator (ARNT) are capable of forming a transcriptionally active heterodimeric complex. The biochemical events that are required for dimerization and transactivation are not fully understood. The purpose of this study was to determine whether covalent modifications of ARNT occur between ARNT existing in the monomeric form and after heterodimerization with the AhR and subsequent binding to DNA. Mouse hepatoma cell line 1c1c7 (Hepa 1) cytosol and ARNT immunoprecipitations were subjected to two-dimensional gel electrophoresis. ARNT was visualized with two antibodies, with distinct epitope specificity, and each detected a considerable level of charge heterogeneity. The pI range observed was 5.7-6.4, with the predominant form at a pI of 6.2. The AhR/ARNT heterodimer was immunoprecipitated from high salt nuclear extract obtained from Hepa 1 cells treated with beta-naphthoflavone using an anti-AhR polyclonal antibody. This immunoprecipitate was subjected to two-dimensional gel electrophoresis, and coimmunoprecipitated ARNT was visualized. The results indicated that ARNT complexed with the AhR in the nucleus has an isoform pattern shifted toward the basic end, with the predominant isoform having a pI of 6.8. Thus, a significant shift in pI occurs during the dimerization and/or after binding to DNA. In vitro transformation of the AhR with 2,3,7,8-tetrachlorodibenzo-p-dioxin in cytosol leads to heterodimerization with ARNT. Two-dimensional gel electrophoresis of ARNT coimmunoprecipitated with the AhR revealed the same isoform pattern as seen in cytosol. This would indicate that each isoform of ARNT is capable of heterodimerizing with the AhRin vitro. ARNT is a phosphoprotein, and the more acidic isoforms appear to have a higher level of phosphorylation. PMID- 9220997 TI - Ligation of RNA-containing duplexes by vaccinia DNA ligase. AB - Vaccinia virus DNA ligase repairs nicked duplex DNA substrates consisting of a 5' phosphate-terminated strand and a 3'-hydroxyl-terminated strand annealed to a bridging template strand. This study addresses the ability of vaccinia DNA ligase to seal nicked substrates containing one or more RNA strands. We found that the viral enzyme rapidly and efficiently joined a 3'-OH RNA to 5'-phosphate DNA when the reacting polynucleotides were annealed to a bridging DNA strand. In contrast, ligation of 3'-OH DNA to 5'-phosphate RNA was slow (0.2% of the rate of RNA-to DNA ligation) and entailed the accumulation of high levels of RNA-adenylate intermediate. A native gel mobility shift assay showed that vaccinia DNA ligase discriminates at the substrate binding step between ligands containing 5' phosphate DNA versus 5'-phosphate RNA at the nick. The enzyme displayed weak activity in RNA-to-RNA ligation on a bridging DNA template (0.01% of RNA-to-DNA activity). Vaccinia DNA ligase was incapable of joining two DNAs annealed on an RNA template. These results can be explained by a requirement for B-form helical conformation on the 5'-phosphate side of the nick. The robust RNA-to-DNA strand joining activity underscores the potential for vaccinia DNA ligase to catalyze RNA-based integration of host cell genetic information into the genome of cytoplasmic poxviruses. PMID- 9220998 TI - [Foreign bodies after total hip prosthesis]. PMID- 9220999 TI - [Association of a low-grade MALT lymphoma and a slightly differentiated adenocarcinoma of the stomach]. AB - The development of synchronous gastric adenocarcinoma and primary gastric lymphoma is rare. We report a case of low grade B-cell lymphoma of mucosa associated lymphoid tissue intermingled with a gastric adenocarcinoma and without Helicobacter pylori infection. This observation leads to discuss the pathogenesis of these tumors and the role of Helicobacter pylori infection in the development of gastric lymphoma and carcinoma. PMID- 9221000 TI - [Juvenile xanthogranuloma localized in the parotid gland]. AB - Juvenile xanthogranuloma of parotid gland is reported in a 9-year-old boy. This kind of tumor is thought to be very rare in salivary glands. Histological, immunohistochemical and ultrastructural data showed characteristic features and excluded a Langerhans cell histiocytosis. Follow-up was uneventful after 18 months. PMID- 9221001 TI - [Neurosyphilis with multiple gummas and AIDS. Report of a case]. AB - The incidence of neurosyphilis has been increasing since AIDS has appeared but acute gummatous necrotizing progression is so far exceptional. We report a second case of "quaternary neurosyphilis" in a 29-year-old patient without serologic nor CT scan evidence of treponema. Diagnosing neurosyphilis is particularly difficult in HIV-1 infected patients, specially in spirochetes-poor injuries. PMID- 9221002 TI - [Pulmonary epithelioid hemangioendothelioma (or IVBAT). Report of a case of exclusively myxoid form]. AB - Epithelioid hemangioendothelioma of the lung, therefore labeled as IntraVascular BronchioAlveolar Tumor (IVBAT), is a tumor of endothelial origin; its presentation with multiple pulmonary nodules radiographically suggests metastatic disease. We describe here an epithelioid hemangioendothelioma with an exclusively myxoid pattern. The tumor was composed of nodules extending in a polypoid fashion from the alveolar septa. The cells within the tumor were epithelial-like and contained cytoplasmic lacunae positive for anti-FVIII, CD31 and CD34 antibodies. According to usual criteria, it was considered as a tumor of low grade malignancy, without vascular or bronchiolar extension or cytonuclear atypia and mitosis; the prognosis, difficult to determine, depends on locoregional tumor progression. PMID- 9221003 TI - [Subcutaneous dorsal tumefaction]. PMID- 9221004 TI - [A tumorous lesion of the thumb in an adolescent]. PMID- 9221005 TI - [An unusual hepatic abscess]. PMID- 9221006 TI - [External evaluation of technical quality of immunohistochemistry. Results of a preliminary multicenter study. Immunohistochemistry Commission of the French Association of Quality Assurance in Pathology and Cytology (AFAQAP-IHC)]. AB - Results of a preliminary study on the evaluation of technical quality in immunohistochemistry are reported. This study has involved 11 centers and has concerned immunodetection of cytokeratins and leucocyte common antigen on paraffin sections. The modalities of slide scoring and the information that can be drawn from the results and from the participants data are discussed. PMID- 9221007 TI - [Immunohistochemical detection of estradiol and progesterone receptors in paraffin sections after treatment with microwaves. Comparison with biochemical assay of receptors in a series of 123 breast cancers with determination of the threshold of optimal positivity]. PMID- 9221008 TI - [Adenovirus and intranuclear inclusions in the appendix in children with acute intussusception]. AB - One hundred and five appendices removed at the time of surgical reduction of intussusception in children were studied by light microscopy after routine procedures to search for aetiological factors involved in intussusception. Normal pediatric appendix specimens served as controls (n = 30). Light microscopic examination showed viral inclusions in epithelial cells in 48 of 105 appendices (45%) from the cases of intussusception. No viral inclusion was observed in controls. Immunohistochemistry performed on 21 appendices from intussusception with a monoclonal antibody against adenovirus showed intranuclear positivity in all appendices with viral inclusions. Viral inclusions seen in epithelial cells of appendices from cases of intussusception are caused by virus and in particularly by adenovirus. The etiological factors involved in intussusception without viral inclusion in appendix remain unknown. PMID- 9221010 TI - The Third Carolina Conference on Tooth Enamel Formation. Chapel Hill, North Carolina, October 18-20, 1995. Conference Proceedings. PMID- 9221009 TI - [Lymphocytic mastitis]. AB - Lymphocytic mastitis is a non infectious inflammatory disease of the breast with lobulocentric lymphocytic infiltrate of variable intensity, collagenous fibrosis and progressive lobular atrophy. The pathogenesis of lymphocytic mastitis is still unknown but some recent reports have suggested an autoimmune origin. We investigated a series of 10 cases by immunohistochemistry and we collected patients' biologic data. The most striking histologic feature was a prominent lobulocentric stromal or intraepithelial lymphocytic infiltrate. Occasionally, the infiltrate was perivascular and nodular along the lobule border. B and T lymphocytes, both demonstrated by immunophenotypic analysis, were shown with a particular pattern of distribution. Pathologists must be aware of this disease in order to recommend immunological investigation. PMID- 9221011 TI - [Anaphylactic or anaphylactoid reactions to paracetamol. Report of 3 cases]. AB - Anaphylaxis to paracetamol exists and perhaps nowadays diagnosis is not confined to clinical evidence of: The repeatability of clinical symptoms. Skin tests. Provocation tests. But also by immuno-biological evidence. Application of the test of Activation of basophils by Flow Cytometry and measurement of leukotriene C4 have been nowadays largely validated and constitute irrefutable proof of the responsibility of the drug as the origin of the clinical symptoms. PMID- 9221013 TI - [International study of prevention. Use of loratadine in the management of children at risk for recurrent respiratory tract infections. Proceedings of a conference. London, 26-28 October 1995]. PMID- 9221012 TI - [Relationship of asthma to gastroesophageal reflux]. AB - A relationship between asthma and gastro-oesophegal reflux (RGO) has been suspected for a long time but never proved. Many studies have shown improvement of asthma by treatment of RGO. The predictive factors of this improvement are unknown. After a short description of the epidemiology of RGO, in general and asthmatic populations, the mechanisms that link the two pathologies have been evaluated, the first consideration being the relationship between bronchial reactivity and RGO, to suggest a practical procedure for asthmatics, with regard to RGO. For therapy, inhibitors of the proton pump arise naturally. PMID- 9221014 TI - [The wheezing child and pediatric respiratory infections]. AB - Given that the respiratory and immunity systems are still in development up to the age of 3 years, viral infections and exposure to environmental factors such as passive smoking in babies and young children may alter these systems, so favouring the development of allergic rhinitis and asthma. This may explain the correlation between viral respiratory infections or exposure to environmental factors and later development of wheezing and asthma. Prevention of respiratory infections may thus reduce the risks of pneumonopathy in the long term. For greatest efficacy however, the preventative measures must be in place before the age of 3 years, i.e. before lung development has been completed. PMID- 9221015 TI - [Respiratory inflammation]. AB - Respiratory virus are the most frequent cause of asthma attacks, and are responsible for more than 80% of episodes of wheezing in children. Atopic subjects have a higher risk of respiratory virus infections, benign or severe, than healthy persons. In children older than 8 years, most respiratory infections are caused by rhinovirus (RV). RV colonizes the respiratory epithelium and provokes a symptomatic rhinitis by non-inflammatory routes (with non involvement of leucocytes). In the nose the most importance of these routes are nerves. In the lower respiratory airways, infection with RV causes an inflammatory reaction with persistence of eosinophils. IL-8 and the other cytokines produced by the infected epithelium extend the action of eosinophils and the inflammatory reaction. The viral/inflammatory pathway is an important new target for development of strategies for the prevention of asthma. PMID- 9221016 TI - [ICAM-1 and allergic inflammation]. AB - Recent progress in the understanding of the physiopathology of asthma makes possible the revision of the objectives of treatment, to pass from short-term symptomatic control to a prolonged control of the illness. L'ICAM-1 is an excellent potential target for preventative treatment of asthma of viral origin. When there are agents available of both inhibiting histamine and positively regulating ICAM-1 there is no longer reason to use simple antihistamines. PMID- 9221017 TI - [Strategy for preventing upper respiratory tract infections. Consensus of the Consulting Committee]. AB - Epidemiological, biological and clinical studies have shown that viral respiratory infections before the age of 3 years play a crucial role in the later development of rhinitis and asthma. Reduction of their frequency and severity may effect the viral alteration of the pulmonary and immune systems. Because of this, as it combines anti-histamine and anti-inflammatory properties, loratadine, a second generation anti-histamine is a candidate for a study as a preventative treatment for allergy and asthma. A clinical study in children of less than 3 years is in progress to establish the value of this product in reduction of the number and severity of infections of the upper airways, the number of associated complications and the incidence of wheezing. PMID- 9221018 TI - [Postnatal development of renal concentrating ability in a desert rodent, Jaculus orientalis]. AB - Postnatal development of renal function was studied in newborn Jerboa from six weeks after birth. The results showed that the renal capacity of urine concentration was more or less analogous to that of the adult, starting 15 weeks after birth when the animal reached a body weight of about 70 to 90 g. From the 13th week onwards, the U/P ratio of the osmolality in function of age, increases when the urinary debit decreases. The osmotic-clearance and the free water clearance will show stability 15 weeks after birth. PMID- 9221019 TI - [What is the current role of clarithromycin in the eradication of Helicobacter pylori?]. PMID- 9221020 TI - [Cytological diagnosis of malignant melanoma of the esophagus]. AB - Malignant melanoma of the esophagus is an extremely rare neoplasm. We presented a case of this neoplasm diagnosed on a cytologic smear from a 84 year-old women. The smear was characterized by scattered dissociated cells with hyperchromatic irregular nuclei, prominent nucleoli and occasional cytoplasmic brown pigment. The immunohistochemical study revealed positivity for HMB-45 and S-100 protein. PMID- 9221021 TI - [Gastric collision tumor with osseous metaplasia]. AB - Neuroendocrine cells are frequently found in gastric tumours, although they rarely make up more than one third of the total number of tumour cells. When juxtapositioning of the two kinds of tumour cells occurs a "collision tumour" is formed. These have been described to occur with varying frequency throughout the digestive tract. They are uncommon in the stomach. We describe a case, of a gastric collision tumour in which an adenocarcinoma coexisted with a carcinoide tumour and there were zones of bony metaplasia in the transition area between the two tumors. Positive CEA, VIP, beta-HCG and TSH on inmunohistochemical analysis was found. PMID- 9221022 TI - [Small intestine adenocarcinoma with Crohn's disease]. AB - We report a 57-year-old male with a 11-years history of Crohn's disease who developed small bowel adenocarcinoma. The patient was admitted with intestinal partial obstruction and he underwent a laparotomy because an abscess in right lower quadrant was suspected. He was successfully treated with resection and adjuvant chemotherapy, without evidence of recurrence after a 2-year follow-up. A review of the literature reveals that about 100 cases have been reported. We describe the distinguishing features of small bowel adenocarcinoma arising in Crohn's disease and we emphasize the difficulty in making an early diagnosis. PMID- 9221023 TI - [Stomach in inguinal-scrotal hernia]. AB - We present two cases of inguinoscrotal hernia that contains stomach, both associated with a chronic obstructive pulmonary disease; one of them is shown with a intermittent gastric obstruction syndrome, the other being a radiological finding in a patient with dispeptic symptoms. The first one died of a respiratory infection, before operation, and the second refused operation because of his scarce symptomatology. A review of the literature shows 60 cases reported including our cases. PMID- 9221024 TI - [Laparoscopic fenestration in the management of symptomatic polycystic liver disease]. AB - We report the case of a 70-year-old woman with polycystic disease of the liver and kidney, complicated with rupture and hemorrhage of the hepatic cysts located in the right lobe. A laparoscopic approach was used. After identifying the polycystic mass, the cytologic examination of the aspirated hematic fluid and the intraoperative biopsy showed its benign nature. An hepatic fenestration was performed. Hemostasis was achieved by electrosurgical and argon probes. The postoperative course was satisfactory and radiological improvement was demonstrated on the CT scan 3 months later. Follow-up examination one year later has been uneventful. Fenestration is one of the less stressing surgical options in the adult polycystic liver disease when treatment is indicated. The laparoscopic approach may be used with safety to perform a similar technique in selected cases, establishing a communication between the cysts and the peritoneal cavity. Nevertheless, it may be difficult to reach deeply situated cysts and to differentiate them from vascular structures. Laparoscopic intraoperative ultrasonography might be of great value in these cases. Likewise, long term studies are needed to determine the benefits and the indications of the laparoscopic fenestration. PMID- 9221025 TI - [Bacteremia for Aeromonas sobria in a patient with cholecystitis]. PMID- 9221026 TI - [Presentation of a case of celiac disease as a hemorrhagic diathesis in an adult]. PMID- 9221027 TI - [Sarcoma in the diverticulum of Meckel]. PMID- 9221028 TI - [Vaginal metastasis of adenocarcinoma of the left colon]. PMID- 9221029 TI - [Dysphagia lusoria caused by an accessory aberrant right subclavian artery. Diagnostic contribution of MR angiography]. PMID- 9221030 TI - Corrigendum to "Molecular cloning of a novel ADP-ribosylation factor (ARF) expressed in planarians" [Biochim. Biophys. Acta 1309 (1996) 205-210]. PMID- 9221031 TI - 4th International Lubeck Conference on the Pathophysiology and Pharmacology of Erythropoietin and other Hematopoietic Growth Factors. Lubeck, Germany, 27-29 June 1997. Abstracts. PMID- 9221032 TI - Behavioral and Psychological Signs and Symptoms of Dementia: Implications for Research and Treatment. Proceedings of an international consensus conference. Lansdowne, Virginia, April 1996. PMID- 9221033 TI - Estrogen and Estrogen-Like Substances in Women's Health: The Year 2000. Proceedings of the 4th International Symposium. Paris, France, October 25, 1996. PMID- 9221034 TI - Introduction to the symposium. PMID- 9221035 TI - New Approaches for Assessing the Etiology and Risks of Developmental Abnormalities from Chemical Exposure. Symposium proceedings. December 11-12, 1995. PMID- 9221036 TI - [New and readmitted cases of tuberculosis reported in the first half of 1996]. AB - Data from TB notification forms, completed between 1.01-30.06.1996, were stored in National TB Database and used to compute TB notification rate structure and the other informations commented here. Yearly notification rate for 1996 was estimated as twice the value for the first 6 months. During the first 6 months, 13488 patients were notified with tuberculosis. Estimated annual notification rate (118.9/100,000) represents an increase of 15.9% as compared to 1995. 87.5% in comparison with 1989 and 113.1% as compared to 1985. Considering TB notifications in 1985 as reference, this increase supplied a cumulative overload of 57438 patients. TB notifications rate shows large differences between districts varying from: 193.3/100,000 (Dolj district) to 43.9/100,000 (Covasna district). Sex differences were also important: 172.9/100,000 in males (301.3/100,000 in Dolj district but 69.8/100,000 in Covasna) and 67.3/100,000 in females (102.9/10,000 in Vranceu but 18.7/100,000 in Convasna). There were large differences in pleural th notification rates (from 32.0/100,000 in Vrancea to 4.3/100,000 in Covasna) and extrapulmonary tb notification rates in various country districts, the lower rates rising the idea of a possible underdiagnosis. 66.9% of notified pulmonary tb cases were smear or culture positive (67.2/100,000), with district values between 97.0% (116.9/100,000) in Mehedinti and 37.3% (39.5/100,000) in Salaj, the lower values could rather be explained by the quality of the sputum examinations than by severity profile of patients. PMID- 9221037 TI - [Eosinophilic pleurisy (a study of 27 cases)]. AB - After reviewing the specific literature mainly on cytokines role in eosinophilic pleural chemotaxis, the study on 27 cases of eosinophilic pleurisy is presented. Only 3 cases were malignant pleurisies. The ratio between blood and pleural fluid eosinophils has no semiologic significance. Tuberculosis and parasitoses have no special significance. Autonomous eosinophilic pleurisies are regarded as a distinct nosologic entity. Their prognosis is favourable. The glucocorticoids are the choice treatment. PMID- 9221038 TI - [An evaluation of the possibilities for a paraclinical study in outpatient pneumophthisiology care and proposals for its improvement]. AB - An attempt was made to make an estimation of equipment and accessibility of pneumophthisiology ambulatory units for diagnosis and treatment of patients newly discovered and the already registered ones. A number of 172 (97% of total number) pneumophthisiology units have been taken into the study showing a certain progress regarding the lb diagnosis. As for other chronic lung diseases, the specific equipment is not available in all units: 73 have not spirometer, bronchoscopy is performed in other places, a.s.o. The situation requires urgent providing the pneumophthisiology network units with new techniques equipment for diagnosis and replacing the present one which is outdated and worn-out. PMID- 9221039 TI - [The prevalence of bronchial asthma, chronic bronchitis and COBP in representative samples of the adult population]. AB - In order to organize the management of asthma bronchiale and chronic bronchitis, in our country, it is compulsory to know the prevalence of these diseases in the autochthonous population. The ECRHS questionnaire (by interview) and lung function tests were applied in three random population samples aged by 18 years and over. The samples were selected from the electoral rolls and coming from three different economic and geographic regions of our country. The questionnaire were administered by trained pneumologists and lung function testing by well trained technicians. The results were stored and processed on computer using the EPI INFO program. The prevalence of asthma bronchiale was: 4.09%, 6.60%, 1.70%. The prevalence of chronic bronchitis was: 12.28%, 10.11%, 7.98%. The prevalence of chronic obstructive bronchitis was: 1.79%, 2.50%, 1.89%. Generally, the prevalence of asthma bronchiale was found close to the values obtained in other studies, of identical conditions. It was particularly close to the results of Italian researchers who studied a population aged by 20 years and over from the northern Italy. The prevalence of chronic bronchitis was near by the value mentioned by Scandinavian epidemiologists. Analytic epidemiology would specify the causes of the variations we founded in the three studied samples. PMID- 9221040 TI - [Serevent in the treatment of moderate bronchial asthma]. AB - A group of 20 patients with moderate bronchial asthma aged 18 through 65, previously treated with Becotide inhaler for 4 weeks was given Serevent in regular dosage for 4 weeks. Serevent efficiency was assessed clinically and functionally (MEVS, IPB). A progressive improvement of all clinical and functional parameters was obtained following the treatment. The evolution was unsatisfactory with 3 cases which entered the group with advanced asthma condition. No adverse reaction of tachyphylaxis manifestations were recorded. Drug adequacy was perfect. The final results place Serevent among elective bronchodilators in long-time ambulatory treatment. PMID- 9221041 TI - [The immunological aspects in the HIV/AIDS-tuberculosis association in children]. AB - HIV-infected subjects are submitted to various immunity disorders, the function of immune competent cells disturbing, the most important one, resulting in severe opportunistic infections. The progressive depletion of CD4 subset of lymphocytes in HIV-infected persons is the most significant one. When HIV infection is associated to tuberculosis there is a more accelerated immune deterioration. The immunological investigation consisted in the determination of cell-mediated immune mechanisms (PPD skin testing, lymphocyte count, subsets of lymphocytes) as well as those humorally mediated (Ig classes, anti-mycobacteria antibodies). PMID- 9221042 TI - [Perinatal HIV transmission. The mechanisms and preventive measures]. PMID- 9221043 TI - [Bronchiolitis obliterans organizing pneumonia (BOOP)--a leading disease in pneumology in the last decade]. PMID- 9221044 TI - [The obstructive sleep apnea syndrome with visceral involvement]. AB - The authors are presenting a SASO of a patient with obesity, hypertension, cardiac ischemic lesions, starting diabetes mellitus, hypoparathyroidism and polypous maxillary sinusitis. Most of this diseases are consequences of respiratory pathology during sleep time; we are discussing in this context the interrelation among different diseases. PMID- 9221045 TI - [The treatment of chronic respiratory insufficiency]. PMID- 9221047 TI - [Home respiratory care. Who cares about it? A report on the Conference of the same name in Ede, Holland, 1-2 Dec. 1995]. PMID- 9221046 TI - [Long-acting beta-2 adrenergic agents--a new therapeutic tool in COBP]. PMID- 9221048 TI - [Tobacco consumption, between the tragedy of the data and indifference]. PMID- 9221049 TI - Dutch presidency has seen a quiet revolution in public health. Interview by Tony Sheldon. PMID- 9221050 TI - [Temporary changes and permanent changes in the erythrocyte blood-group antigens in malignant hemopathies]. AB - Samples of peripheral blood were taken from 11 patients (blood group A, B and O), suffering from acute myeloblastic leukemia (AML), acute lymphoblastic leukemia (ALL), myelodysplastic syndrome (MDS), before and after chemotherapy, compared to normal control samples. The RBC's typing was done by standard agglutination technique (with conventional human and murine anti-sera) and flow cytometry. While using different reagent dilutions, a lower expression of the A or B antigen was noticed in all patients, even if direct typing of the RBC's revealed an apparently normal pattern. The most important depletion of antigenic expression was found to correspond to the highest concentration of myeloblasts in the bone marrow, with hypoplastic erythrocytic series. The modified H reactivity, detected at admission, was still present after complete remission, as an expression of the residual disease. Studies of the H expression could eventually become a parameter of evaluating the moment of relapse of the myeloproliferative disease. PMID- 9221051 TI - [Academic eulogy of Professor Henri Gastaut--foreign honorary member]. PMID- 9221052 TI - [Human factors and heart surgery: a Cartesian dream]. AB - It is postulated that high technology medicine can be assimilated to complex socio-technical systems such as the aviation industry, nuclear power or chemical plants etc. It is proposed to apply to cardiac surgery the techniques of human reliability and human error analysis that have been acquired over the past two decades to enhance safety in those areas of high technology. It is now widely accepted that in complex socio-technical systems accidents are due to human factors in 60-80% of the cases. Accident theories and, in particular theories of organisational accidents, have been applied prospectively to negative surgical outcomes in an attempt to understand their causation ad to establish defence mechanisms to prevent them or at least mitigate their consequences. The philosophical issues raised by this endeavour will be outlined. PMID- 9221053 TI - [Insulin resistance: lessons from animal models of obesity]. AB - The insulin resistance of animal models of obesity (the gold thioglucose obese mouse and the o b/o b mouse) is characterized by several abnormalities. At the receptor step, both the binding function (decreased number of sites) and the enzymatic, tyrosine kinase function (decreased insulin activation) are altered. At postreceptor steps, phosphatidylinositol 3-kinase (PI3-K) plays an important role in insulin signalling, particularly for the stimulation of glucose transport in muscle and adipocyte. Insulin activation of PI3-K is markedly diminished in obese mice; starving the obese animals restores normal responses of PI3-K, glucose transport, and glycogen synthesis, to insulin. These observations emphasize the multi-site, and largely reversible, nature of insulin resistance in these animal models of obesity. Similar alterations have been reported in the literature with regard to the sites of insulin resistance in human obesity and non insulin-dependent diabetes. PMID- 9221054 TI - [Non-insulin-dependent diabetes: from physiopathology to treatment]. AB - Non-insulin-dependent (or type 2) diabetes mellitus is a common, underdiagnosed and growing disease in our society. It is responsible for increased morbidity and mortality and represents an important public health problem. This polygenic disease is often expressed late in life and its evolution is accelerated by environmental factors leading to obesity. It combines defects in both insulin secretion and insulin action, and such defects are present in various proportions according to the type of patient and the stage of the disease. Diet and physical activity recommendations are the basis of the treatment. Current pharmacological approaches aim at improving insulin secretion and/or insulin cellular action. After secondary failure to oral drugs, insulin therapy should be initiated, the patient becoming "insulin-requiring". A synergy should be searched in the combination of various therapeutic modalities in order to improve the glycaemic control. PMID- 9221055 TI - Proceedings of the 7th International Conference on Hand-Arm Vibration. Prague, May 9-12, 1995. PMID- 9221056 TI - [Musical excerpts: indices relating to familiarity, age of acquisition and verbal associations]. AB - The objective of the present study was to establish estimates of familiarity, of age of acquisition, and of verbal associations in relation to 144 musical excerpts drawn from the repertoire of tunes that is expected to be shared by all French-speaking Quebec university students. The excerpts were synthesized monophonic lines (which can be found in Appendix B) that were tape recorded. A first group of 60 university students were required to indicate their degree of familiarity (on a 5-point scale) with each excerpt and the age period at which they learned the excerpt. A second group of 60 students indicated whether the original tune was vocal or instrumental as well as the first words that came to mind. In each group, half the subjects were presented with the 144 excerpts in a different order. Overall, the material was found to be highly familiar and to have been mostly learned between the ages of 5 and 15. Most of the excerpts were easily categorized as vocal or instrumental with the exception of 26 excerpts, which were accordingly classified as ambiguous. Finally, 57% of the excerpts elicited verbal responses. There was a high level of agreement in some of the verbal responses provided, referred to as dominant responses. In contrast, 21 excerpts gave rise to very little verbal recall, hence specifying a subset of "purely" musical material. All indices of familiarity, of acquisition age, and of verbal responses (specifying the overall rate of verbal responding, the content of the dominant response as well as its frequency of occurrence) are provided for each excerpt in Appendix A. Finally, the most interesting aspect of the present study was revealed by the analysis of the errors in verbal recall. Subjects tended to fill in missing elements by words that fitted the meaning of song lyrics as well as the temporal structure of the music. For instance, instead of providing "MON BEAU SAPIN", subjects would provide "mon grand sapin". The use of such a procedure is consistent with the notion that music serves as a memory aid which facilitates the communication of news and ideas. PMID- 9221057 TI - [The role of quinine chlorhydrate in the conditioned inhibition of the tarsal reflex in Drosophila melanogaster]. AB - The proboscis extension reflex (PER) can be elicited by applying a sucrose stimulation to the tarsus of a walking fly. This reflex decreases in frequency with repetition, presumably habituation, a nonassociative learning. If each sucrose stimulation is followed by a bitter stimulation, quinine chloride, the PER declines more rapidly, probably the result of conditioning, an associative learning. The present work shows that quinine chloride does not always inhibit PER suppressions but depends on the moment of delivery, being most effective when presented immediately after a sucrose stimulation. A bitter stimulus presented before, or simultaneously with a sucrose stimulation is less effective than habituation to sucrose alone. This experiment provides evidence for an interpretation in terms of cognitive association. The model of learning is not a Pavlovian conditioning as advanced by Medioni and Vaysse (1975), but corresponds to the punishment paradigm of Dyal and Corning (1973). PMID- 9221058 TI - [The induction of mutations in Streptomyces aureofaciens by UV rays and nitrosoguanidine and the identification of the mutants obtained]. AB - Streptomyces aureofaciens 019(8), chlortetracycline producent was subjected to the effect of UV-light and nitrosoguanidine and after that its 121520 colonies have been analyzed by the methods of replicas and 57 mutants were isolated: sensitive to the effect of UV radiation--14; auxotrophs--30; antibiotic-inactive -13. The induction frequency of mutations is within the limits of 0.016 to 0.067%. Auxotrophic mutants manifested genetic instability. Mutants with a block chlortetracyclin biosynthesis are stable. PMID- 9221059 TI - [The effect of a modification to the lipopolysaccharide of Yersinia pestis on its neutrophilokine-inducing activity]. AB - The ability of the isolated and modified Yersinia pestis LPS to induce the synthesis of the neutrophilokines that regulate the macrophage functional activity was studied. It is established that the Y. pestis LPS detoxication, especially by the method of deacylation, does not lead to the decrease in biological, in particular neutrophilokine-inducing activity of these preparations, but actually even increases it. These results are in agreement with many reports showing the possibility of decreasing the LPS toxicity without reducing immunostimulatory activity of this important component of the outer membrane of gram-negative bacteria. PMID- 9221060 TI - [The isolation of the antigenic substances of Mycobacterium tuberculosis and the characteristics of their basic chemical components]. AB - Different methods of isolation were used when obtaining the antigen determinants of the tuberculosis mycobacteria (tuberculins, polysaccharide, phosphatide). Main chemical components of the antigens were investigated. Most effective and economic methods of obtaining the M. tuberculosis antigens substances were chosen. The using of the obtained antigens in the test systems for diagnostics of tuberculosis discussed. PMID- 9221061 TI - [The immunosuppressive activity of filtrates of the culture broth from nonvirulent salmonellae]. AB - Cultural filtrates (CF) of avirulent Salmonella virchow and Salmonella dublin have been studied for their influence on the delayed type hypersensitivity (DTH) to test-antigen in mice. It is found that i.p. injection of CF inhibits DTH in mice. The immunosuppressive activity is indicated in lipid fraction of CF and it may be removed from CF by the O-specific immunosorbent. It is heat-stable and disappears after phenol or trichloroacetic acid treatment. Gel filtration data evidence for high molecular weight of the active factor. These facts indicate to the connection of immunosuppressive activity with native lipopolysaccharide of avirulent Salmonella. PMID- 9221062 TI - [The metabolism and bactericidal properties of the blood neutrophilic granulocytes in experimental typhoid intoxication]. AB - The levels of succinate dehydrogenase (SDG), lactate dehydrogenase (LDG), glucose 6-phosphate-dehydrogenase (G-6-PDG), myeloperoxydase (MPO), glycogen and cationic proteins were determined in the neutrophilic granulocytes of peripheral blood of 79 rabbits after experimental contamination by endotoxin of S. typhi. The bactericidal system of neutrophils was stimulated due to the depression of SDG, activation of LDG and G-6-PDG levels. Administration of indometacinum and chlotasolum blockaded the cyclooxygenase fermentative system of prostaglandin synthesis and thus decreased the activity of S. typhi endotoxin. PMID- 9221063 TI - [The distribution of leptospirae in the icterohaemorrhagiae serogroup]. AB - The capacity of leptospiras to get acclimated in the organism of unusual hosts has been studied. Cultures of Icterohaemorrhagiae from musquashes, house mice and field voles have been isolated of leptospirosis sources under intensive epizootics among rats. A possibility to reproduce Leptospira carrying by the cultures of leptospirae of serogroup icterohaemorrhagiae in musquashes and water voles has been shown. One can conclude that under icterohemorrhagic leptospirosis musquashes and water voles can take the part of additional source of infection but they cannot independently preserve leptospirae in nature as a biological species grey rats being their main hosts. PMID- 9221064 TI - [Microphytic algae in the pathology of the cetaceans]. AB - The review includes the analysis of the world literature (1913-1995) about the problems of interrelations, between microorganisms (microphytic algae and cyanobacteria) and marine mammals (whales and dolphins). Special attention is payed to the parasitological, bioindicative and toxicological aspects of microalgae inhabiting the surface of the skin integuments and respiratory organs of cetaceans, as well as their environment. There is the list of the algal species overgrowing the animals bodies and their hosts. The most of the epibionts are belonging to diatoms, there are also some cyanobacteria and green algae. The probability of the influence of the algological factor on the hosts health and a possibility of the microalgae use to evaluate the status of animals and their environment have been discussed. PMID- 9221065 TI - Response to "Determinants of Decision Making for Circumcision" by C. Cielsielski Carlucci, N. Milliken, and N.H. Cohen (CQ, Vol 5, No 2).Circumcision: ethical and human rights impact assessment. PMID- 9221066 TI - 9th International MASCC Symposium on Supportive Care in Cancer. St. Gallen, Switzerland, 26 February-1 March 1997. Abstracts. PMID- 9221067 TI - [Cytological diagnosis of voluminous processes of the liver]. PMID- 9221068 TI - [A simple method of measuring ornithine decarboxylase activity in mixed human saliva]. AB - A simple and sensitive method for measuring ornithine decarboxylase in mixed human saliva is described, based on assessing the waste of the reaction substrate ornithine by modified Chinard's color reaction. The highest specific activity of the enzyme (up to 1.5 ncat/ml) is found in salivary samples collected on an empty stomach in the morning. After centrifugation of the saliva the entire ornithine decarboxylase activity is localized in the mucin sediment. PMID- 9221069 TI - [Gas chromatographic analysis of lipids in exhaled air condensate in children with bronchopulmonary diseases]. AB - The lipid fatty-acid spectrum in exhaled air condensate (EAC) was analyzed in children with bronchopulmonary diseases by gas chromatography. The detected shifts in the fatty-acid composition of EAC lipids may be used for validating a therapeutic approach and monitoring the treatment efficacy. PMID- 9221070 TI - [Method of measuring non-protein bound fucose]. AB - A method for measuring fucose not bound to proteins in the serum is developed. The level of this enzyme is shown to increase with age. It is increased in adult patients with myeloma, liver cirrhosis, and rheumatic fever and in children with exacerbations of chronic osteomyelitis and gastroduodenitis. PMID- 9221071 TI - [High performance liquid chromatography in the analysis of amino acids contents of the stomach and duodenum]. AB - High-pressure liquid chromatography was used for measuring the content of amino acids in various portions of the gastrointestinal tract and for correcting the composition thereof using carbon adsorbents of different composition. Standard Beckman equipment and methods routinely used for analysis of plasma and serum were used. The data are well reproducible and permit us to recommend this highly sensitive method for investigating amino acid metabolism and for its correction in the digestive tract. PMID- 9221072 TI - [Development of monoclonal antibody-based immunoenzyme kit for detection of human thyrotropin hormone]. AB - An enzyme immunoassay kit based on monoclonal antibodies (Mab) has been developed for measuring thyroid-stimulating hormone in human serum and plasma. Laboratory trials demonstrated its high sensitivity (3.1 microIU/ml), specificity, reliability, and accuracy. The analysis is simple and rapid (2.5 h), which is convenient for clinical use. The kit permits differentiation between normal subjects and patients with primary hypo- or hyperthyroidism and helps monitor the treatment efficacy. Comparison with the Amerlite TSH-60 kit, carried out on 357 sera, showed a high correlation coefficient (0.96) between the two kits. PMID- 9221073 TI - [Diagnostic test kit for detection of antibodies to syphilis agent]. AB - A diagnostic enzyme immunoassay test kit is proposed for detecting anti-treponema antibodies in sera of syphilis patients, usable in two variants: on prefabricated polystyrene plates or by immunofiltration using a membrane filtration enzyme immunoassay kit. A peculiar feature of the test kit is the use of enzyme immunoassay conjugate based on monoclonal antibodies to light chains of human immunoglobulins, highly purified antigen from cultural T. pallidum, two substrates for detecting enzymatic activity. The efficacy of the test was confirmed on 163 patients' and control sera and compared with that of the complement fixation test, two modifications of immunofluorescent test, and micromodification of the precipitation test. The test proved to be highly sensitive, specific, and reproducible, and is recommended as a confirmation test for the diagnosis of syphilis. PMID- 9221074 TI - [Laboratory diagnosis of iron metabolism disorders (lecture)]. PMID- 9221075 TI - [Diagnostic significance of measuring levels of large granule-containing lymphocytes as a screening test]. AB - The efficacy of measurements of large granule-containing lymphocytes (LGL) as a screening test for population screening has been certified. Liquidators of the Chernobyl accident aftermath in 1986 and 1987, residents of the Tomsk district exposed to radiation as a result of accident, and patients with general somatic and malignant diseases were examined. The LGL level was fit for dividing the examinees into healthy and ill, but not for selecting subjects at a high risk of cancer. The reaction of the natural killer system to endogenic and exogenic injury is similar. PMID- 9221076 TI - [Proliferative activity of lymphocytes in a culture with phytohemagglutinin and immunity parameters in acute pneumonia]. AB - Proliferative activity of lymphocytes in a culture was assessed by flow cytometry and lymphocyte blast transformation test (LBTT) with the proliferative pool of cells (S+G2+M) from 150 patients with acute pneumonia, 30 of these with the mild form, 97 with medium-severe, and 27 with grave form; in 43 patients the disease duration was over 30 days (protracted pneumonia). In addition, levels of R.protein, homoreactant, immunoglobulins, complement components, T, B, and active T lymphocytes, phagocytosis, and granulocyte oxidative metabolism were assessed. LBTT values were in inverse relationship with the disease severity. Correlations between the studied immunity parameters were analyzed. The findings indicate that LBTT is an informative method for assessing the immune reactivity and may serve for predicting the disease course and for the differential diagnosis of pulmonary diseases. PMID- 9221077 TI - [Comparative incidence of detection of specific antibodies to Yersinia pestis capsular antigen and lipopolysaccharide in humans immunized with pest vaccine]. AB - Two to 8 years after vaccination, specific antibodies to capsular antigen (F1) and lipopolysaccharide (LPS) are found in the blood sera of but 10 to 20% of subjects 1-4 times epicutaneously immunized with live antipest vaccine. Only regular continuous (for at least 7-15 years) antipest vaccination leads to the production of antibodies to the main antigens of Yersinia pestis, detectable in remote periods in 75.9 +/- 4.5% of vaccinees. It is noteworthy that the number of positive responses and titer of antibodies to LPS is appreciably higher than to F1. Detection of antibodies to Y. pestis is recommended for the retrospective diagnosis of pestis in both humans and animals in natural foci of this infection. Competitive enzyme immunoassay on nitrocellulose membranes with monoclonal antibodies is to be preferred for the purpose. PMID- 9221078 TI - [Gastrointestinal probe]. PMID- 9221079 TI - [Micromethod of measuring peroxide chemiluminescence of blood plasma]. PMID- 9221080 TI - [Normal values of the de Ritis coefficient]. PMID- 9221081 TI - [Reagent kits for clinical laboratory diagnosis: the developing and registration system]. PMID- 9221082 TI - [Leading specialists of the Ministry of Public Health and Medical Industry of the Russian Federation]. PMID- 9221083 TI - [Approval of the list of organizations, enterprises, works and their structural divisions whose activities grant the right to fix an extra 20% payment (wage rate) for diagnosing and treating HIV-infected persons and also for work connected with HIV-containing specimens]. PMID- 9221084 TI - [Working hours and vacations of specialists dealing with the diagnosis and treatment of HIV-infected persons and those working with HIV-containing specimens]. PMID- 9221085 TI - [State of intracellular components of the neutrophil bactericidal system in patients with cholecystitis]. PMID- 9221086 TI - [Cytochemical method of simultaneous determination of oxidation-reduction enzymes and acid phosphatase in blood lymphocytes]. AB - The results of cytochemical tests for simultaneous measurements of dehydrogenases and acid phosphatase in blood leukocytes do not differ from the results of the original methods developed by R. P. Nartsissov (1969) and A. Goldberg and T. Bark (1962). Use of this method permits the physician to have simultaneous information about two aspects of metabolism in the cells and cuts down the time needed for measuring the activities of two enzymes in the leukocytes. PMID- 9221087 TI - [Use of diffractometry of blood smears in the diagnosis of rheological disorders and assessment of the course of blood system diseases]. PMID- 9221088 TI - [Medicinal plants from the viewpoint of nations of the European Union]. AB - Vegetable drugs and preparations manufactured from the represent an important commodity in the world market and their production is on the increase. The problems in the individual countries, different opinions of their definition, evaluation utilization and distribution are discussed. PMID- 9221089 TI - [Materia medica in ancient Indian medicine. Two of the oldest Indian medical manuscripts. I]. PMID- 9221090 TI - [Pharmacoeconomics. I]. AB - The present paper aims to define the meaning of the concepts pharmacoeconomic research and pharmacoeconomics. It presents brief characteristics of the individual types of analyses cost minimization analysis, cost-of-illness, cost benefit analysis, cost effectiveness analysis, and quality of life analysis. PMID- 9221091 TI - CQI in health care: some comments on "can it really work"? PMID- 9221092 TI - Integrating CQI in health organizations: some perspectives. PMID- 9221093 TI - Sustaining CQI. PMID- 9221094 TI - "Every defect is a treasure." Quality problems that result in adverse outcomes provide opportunities for quality improvement. PMID- 9221095 TI - Proceedings of the 2nd International Symposium and Workshop on Biological Environmental Specimen Banking. Stockholm, Sweden, May 20-23, 1996. PMID- 9221096 TI - Proceedings of the Wolf Creek IV Conference on Cardiopulmonary Resuscitation. Palm Springs, California, April 14-16, 1996. PMID- 9221097 TI - [Diagnosis of drug allergy]. PMID- 9221098 TI - [Exposure to pollutants and allergens in the asthmatic child compared with the healthy child]. AB - To assess the frequency of exposure to allergens and indoor pollutants of school age asthmatic children at the time of their first visit to the specialist, we studied 14 cases with the diagnosis of asthma according to international criteria, and 21 healthy controls. The parents of the children filed a questionnaire asking about socio-economic level, family history of asthma, exposure to allergens or indoor pollutants, and clinical severity of the disease. Questionnaires with less than 80% of the responses were excluded from analysis. Asthmatic patients had higher frequency of exposure to tobacco smoke (42.8% vs 38%), moisture in the home walls (42.9% vs 19%), and to dust reservoirs (71.4% vs 52.4%). A high proportion of the asthmatic patients did not apply adequate environmental control measures. Education for the patients and their primary care physicians must be increased, to reduce the morbidity of the diseases. PMID- 9221099 TI - [Urticaria]. AB - The urticaria common sickness is a multifactorial etiology which in a lot of occasions can't be determined in spite of the exhaustive investigation of treatment doesn't solve definitely the symptomatology, because of they have protocols of investigation research that orients us to find the common causes , the diagnostic methods to follow in each case and of this trained or form offer effective treatment that gain relief in definitive the sickness. This revision make mention of the classification etiological of the urticaria, history concepts, frequency, histopathology, chemical mediator and immunologist that participate in and the evaluation diagnostic primary to realize in the patient with urticaria acute and chronic. PMID- 9221100 TI - [Vasculitis and viral infection]. AB - Viruses have been implicated in vasculitis. To determine activity of viral infection associated with vasculitis. 17 patients with vasculitis had been in immunological and antiviral antibodies evaluation. Twenty five healthy controls sex and age matched with hematic biometry (BH) and AA. All subjects were negative to HIV and HBV. Viral activity was demonstrated in eight patients; vascular purpura (5), Takayasu disease (1), polyarteritis nodosa (1), erythema nodosum (1). None subject of control group had IgM activity. Antibodies response of IgG in patients were of lesser intensity than in control group. 14 abnormalities in BH were found in patients and 4 in control group. Immune response in patients, measured by lymphocyte subpopulations and circulating immune complexes was abnormal. In conclusion 47% showed viral activity, but the dominant feature was abnormal immune response in 82%. PMID- 9221101 TI - [Controlled-delivery drugs]. AB - The possibility of using modern means for systemic therapy, requires methods of delivery with the ability to modify such delivery once it has started. Some of these methods are already available as is the case of patches for a variety of indications, providing the passage of the medicine through the skin. Other devices, more complicated and of wide clinical use, may enable controlled liberation of substances directed to specific organs. A brief review of these systems is presented. PMID- 9221103 TI - [Desensitization to antibiotics]. AB - It is communicated an alternative to value and take experience in the molecules behavior that are expressed biochemically with haptens. It is explained the reason of the practical application of the dilution of the medicine that unfettered allergy when is used as antigen for prick test. It is mentioned the form in which is systematized the application of the medicine that produces allergy. The measurement method of the risk-benefit is safe practical and of physiological present time and biochemistry. PMID- 9221102 TI - [Measurement of sIgA in patients with chronic stable bronchitis]. AB - As we know secretory IgA of respiratory system has a very important role in defense mechanism. We studied 100 human beings, 50 healthy persons and 50 chronic bronchitis patients. Lavage nasal samples were tacked from healthy persons and sputum samples from chronic bronchitis patients. The laboratory test was nefelometry laser. Samples were analyzed was 1 student. Our results showed light increased of IgA in chronic bronchitis patients not significative. We concluded that this light increased is secondary to continuous stimulus of bronchial mucous as a part of defense mechanism. PMID- 9221104 TI - American Spinal Injury Association annual meeting abstracts. PMID- 9221105 TI - [Calcium transport in microsomes isolated from rat parotid gland. Effects of ADP and an ATP-regenerating system]. AB - Calcium loading of a rat parotid microsomal fraction is greatly increased by an ATP-regenerating system (phosphocreatine and creatine phosphokinase). This effect is neither a consequence of a rise in the ATP concentration nor of an increased formation of inorganic phosphate originating from hydrolysis of ATP or phosphocreatine. Addition of ADP to the incubation medium provokes an inhibition of Ca2+ influx and a stimulation of Ca2+ efflux by the microsomal fraction. These results suggest that the stimulation of Ca2+ uptake by the ATP-regenerating system is due, at least in part, to an increase of Ca2+ influx and a slowing down of Ca2+ efflux as consequence of a decrease of ADP availability. It is proposed that the effect of ADP on Ca2+ movements could account for the action of certain agonists on intracellular Ca2+ concentration. Moreover, the InsP3 responsive Ca2+ pool was also shown to be enlarged by the ATP-regenerating system without modification of InsP3 sensitivity. PMID- 9221106 TI - The powers and perils of e-mail. PMID- 9221107 TI - Noise adaptation in integrate-and fire neurons. AB - The statistical spiking response of an ensemble of identically prepared stochastic integrate-and-fire neurons to a rectangular input current plus gaussian white noise is analyzed. It is shown that, on average, integrate-and fire neurons adapt to the root-mean-square noise level of their input. This phenomenon is referred to as noise adaptation. Noise adaptation is characterized by a decrease in the average neural firing rate and an accompanying decrease in the average value of the generator potential, both of which can be attributed to noise-induced resets of the generator potential mediated by the integrate-and fire mechanism. A quantitative theory of noise adaptation in stochastic integrate and-fire neurons is developed. It is shown that integrate-and-fire neurons, on average, produce transient spiking activity whenever there is an increase in the level of their input noise. This transient noise response is either reduced or eliminated over time, depending on the parameters of the model neuron. Analytical methods are used to prove that nonleaky integrate-and-fire neurons totally adapt to any constant input noise level, in the sense that their asymptotic spiking rates are independent of the magnitude of their input noise. For leaky integrate and-fire neurons, the long-run noise adaptation is not total, but the response to noise is partially eliminated. Expressions for the probability density function of the generator potential and the first two moments of the potential distribution are derived for the particular case of a nonleaky neuron driven by gaussian white noise of mean zero and constant variance. The functional significance of noise adaptation for the performance of networks comprising integrate-and-fire neurons is discussed. PMID- 9221108 TI - The joint development of orientation and ocular dominance: role of constraints. AB - Correlation-based learning (CBL) has been suggested as the mechanism that underlies the development of simple-cell receptive fields in the primary visual cortex of cats, including orientation preference (OR) and ocular dominance (OD) (Linsker, 1986; Miller, Keller, & Stryker, 1989). CBL has been applied successfully to the development of OR and OD individually (Miller, Keller, & Stryker, 1989; Miller, 1994; Miyashita & Tanaka, 1991; Erwin, Obermayer, & Schulten, 1995), but the conditions for their joint development have not been studied (but see Erwin & Miller, 1995, for independent work on the same question) in contrast to competitive Hebbian models (Obermayer, Blasdel, & Schulten, 1992). In this article, we provide insight into why this has been the case: OR and OD decouple in symmetric CBL models, and a joint development of OR and OD is possible only in a parameter regime that depends on nonlinear mechanisms. PMID- 9221109 TI - [Bile secretion and the nuclear transcriptional activity of rat liver cells under the action of bile acids]. AB - The intensity of rat's bile secretion under the intraportal infusion of cholic and taurocholic acids has been studied. The results demonstrate that bile acids decrease bile secretion rate under the infusion of these acids in the physiological saline, and increase bile secretion rate under the infusion in saline, with hydrocarbonic ions. Previous treatment by the transcriptional inhibitor actynomycin D affected the development of the hypercholeretic action of the taurocholic acid. The transcriptional activity of the isolated nuclei of liver cells is increased under taurocholic acid action, and decreased under the cholic acid action. The results show that choleretic effects of the bile acids is realised, at least, partially, on the level of the regulatory molecular mechanisms of the cell, the transcription among them. PMID- 9221110 TI - [The antidiphtheria immunity of the eligible donor population]. AB - Antidiphtheritic antibodies were studied in the blood serum of Kiev and Kiev region population capable of blood donation (984 persons). It has been found out that antidiphtheritic antibodies were present in men's and women's blood serum of different blood groups (by ABO system) during a year. Antidiphtheritic antibodies with high titers > 1.80 were most often found in autumn, winter and spring in people having A (II), B (III) and AB (IV) blood groups. PMID- 9221111 TI - [The typological characteristics of the stress activation of lipid peroxidation and its correction by thymopentin]. AB - On the model of acute stress in rats the dependence of accumulating of peroxide hydrogen from typological peculiarities of organism was determined. The highest level of activation of lipid peroxide oxidation coincides with maximally expressed impairing of stomach mucous membrane the dependence of processes of lipid peroxide oxidation in stress from typological peculiarities of organism was grounded as well as high effectiveness of correction of ulcerogenic effects by neuropeptide thymopentin. PMID- 9221112 TI - [The physiological properties of the action of a new analgesic and antipyretic preparation]. AB - Children's morbidity from Chernobyl accident aftereffects was studied. New preparation "Paravit" was created. This preparation is for treatment illnesses, which are accompanied by pain and slight temperature. Tablets "Paravit" have mark, which allows exact dosing for children of different age. The specific pharmacological investigation of "Paravit" were taken. They showed that new preparation possesses active antipyretic and analgetic properties. PMID- 9221113 TI - [Pharmacological modulation of the cholecystokinetic action of Naftusia water]. AB - In clinical-physiological research with using method of cholecystoultrasonography it is shown, that the most perceptibly increase stimulating action water "Naftusia" on postprandial emptying of gallbladder by its hyperkinesia alpha adrenoblockers and gastrozepin, and the most effective pharmacons braking stimulation by water hyper- or normokinetic postprandial reaction of gallbladder may consider H2-blockers and platyphyllin. PMID- 9221114 TI - [The effect of dimethylethanolamine on the summation capacity of the central nervous system and on the work capacity of animals in a chronic experiment]. AB - The effects of inhaling action of dimethylethanolamine (twenty-four-hour for four months (on summarizing liminal index, SLI) and efficiency of white rats in dependence on various concentrations of amino alcohol were studied in chronic experiments. The obtained results allowed to conclude, that high (2.76 mg/m3) concentration of dimethylethanolamine influenced on functional state of central nervous system. SLI changes pointed to disturbance of dynamic equilibrium between processes of inhibition and excitation with prevalence of latter. In the same time the sufficient grounds for the attribution of dimethylethanolamine to myorelaxants were absent, since in our experiments we used only very high concentrations of this agent. PMID- 9221115 TI - [An analysis of the seasonal differences in the reaction of the hypothalamo hypophyseal-ovarian system to the light regimen of the housing of animals after destruction of the lateral septal nucleus]. AB - The investigation of the lateral septal nucleus (LSN) destruction participation in biorhythmological processes of the morphofunctional condition of the female genital apparatus of female rats was made in the experiments of juvenile female rats, i.e. such which are caused by the change of seasons of the year and light condition. It is shown that the reaction of the hypothalamo-hypophysis-ovary system (HHOS) on the changes of light conditions depends on the season of the year: in spring the regular darkness lends to the negative consequences in the sexual system, and in summer and autumn on the contrary, makes in more active. Regular lighting stimulates the HHOS only in spring. The realisation of photoperiodic processes in HHOS may be possible only in the presence of intact LSN. It is exposed that the reaction of HHOS of animals with destructive LSN in all seasons of the year and under different light conditions are opposite to those which take place in female rats with intact LSN. PMID- 9221116 TI - [The role of the endothelium and of biologically active substances of endothelial origin in regulating blood circulation and cardiac activity]. AB - The role of endothelium and its biologically active derivatives in the central and local control of circulation is under consideration. Molecular and cellular mechanisms of the activation of the endothelium-dependent responses of different functional significance are being discussed, as well as the state of endothelial responses in the development and compensation of pathological processes in the cardiovascular system. PMID- 9221117 TI - [The neurophysiological effects of the beta-blocker obzidan in the modelling of emotional stress]. AB - Neurophysiological effects of beta-blocker obsidan have been studied in experiments with 3 month old male Wistar rats under conditions of modelling of emotional disturbances. It is shown that the 7-8 day stress induced profound changes in electrographic, emotional and vegetation reactions of rats. Disturbances induced by the stress are observed for a long-term poststress period. Controlling and normalizing the level of excitability of the cortex subcortex structures the beta-blocker obsidan promotes reversibility of the stress-induced changes. Central beta-adrenoreactive systems of the brain play the important role in mechanisms of formation of stress states and their consequences. PMID- 9221118 TI - [The role of potassium and chlorine ions in the gas-transport function of the erythrocytes]. AB - The research of electrolytes influence on blood gas-transport function was carried out on blood samples taken from human finger. It was shown that in the absence of blood system diseases, with pH indices, hemoglobin content, HCO3-, Na+ concentration in blood within physiological norm, partial pressure of oxygen and hemoglobin saturation by oxygen is clearly determined by concentration gradient of chlorine ions on erythrocytes membrane, that is by oxygen transport through the erythrocyte membrane, which is attended by the transfer of chlorine ions. Under these conditions there are two levels of PO2 indices and hemoglobin saturation by oxygen, depending on the concentration gradient of chlorine ions on the erythrocyte membrane, that are determined by K(+)- and Cl(-)-ions concentrations level in the blood and erythrocytes. PMID- 9221119 TI - [The influence of the limbic cortex and hypothalamus on the impulse activity of the bulbar respiratory neurons and on respiration under hypoxia]. AB - The response of bulbar respiratory neurons and the total inspiration to stimulation of the limbic cortex and hypothalamus was not identical as a result of different sensitivity of the studied structures. The hypothalamus exerts mainly facilitating influence both in norm and at the maximal altitude (7500-8000 m). The limbic cortex exerts mainly inhibitory influence. At the maximal altitude no typical reactions of the respiratory neurons and the total respiration were observed in response to stimulation. PMID- 9221121 TI - [Changes in the level of gamma-aminobutyric acid in the blood of children with thyroid diseases]. AB - Blood GABA level was studied in children with thyroid disorders. Increase of GABA level was revealed in thyroid hyperplasia (11 degrees) and euthyroid goiters; in blood of children with diffuse toxic goiter there changes are much more significant. In children with thyroid cancer dramatic increase of GABA content was observed; in the nearest time following thyroidectomy blood GABA level decreased to low values, several mouths later it became normal, in a year and more it became elevated again. In blood from three children with congenital hypothyrosis the level of the mediator was decreased; no changes in GABA level were observed in children with autoimmune thyroiditis. Two hours after falling asleep blood GABA content lowered both in normal children and those with euthyroid goiter and thyroid cancer. The mediator level elevated sharply after finishing physical exercises in normal children and those with euthyroid goiter, while in children with thyroid cancer response of GABAergic system to physical exercises was opposite and in operated children it was absent. PMID- 9221120 TI - [The contractile reactions of vascular smooth muscles in hypercholesterolemia and changes in the functional activity of the endothelium]. AB - Experiments on the isolated stripes of the portal vein of rabbits in experimental hypercholesterolemia and/or inhibition of endothelium's synthetic function with NO-synthase inhibitor L-NAME revealed the decrease of the contractile responses of vascular smooth muscles to stretching, as well as endothelium-dependent relaxation of aortal stripes to acetylcholine. L-arginine stimulated the synthetic activity of endothelium, thus enabling to use it for the correction of functional disturbances of the endothelium and of the studies responses. PMID- 9221122 TI - [Circulatory changes in acute myocardial ischemia in dogs with experimental diabetes mellitus]. AB - On alloxane-diabetic dogs under chloralose anaesthesia without opening the chest catheterization, extracorporal perfusion and resistography of coronary arteries, catheterization and continuous drainage of coronary sinus, catheterization of major vessels and heart chambers were performed. Acute myocardial ischemia was induced by the 60 s cessation of left circumflex coronary artery extracorporal perfusion. The magnitude and peculiarity of the systemic circulation reactions during acute myocardial ischemia in dogs with moderate and mild hyperglycemia (less than 12 mmol/l), didn't differ from those in control group. But the degrees of coronary arteries dilation in the ischemic area and coronary sinus blood oxygen saturation reduction were less and the velocity of the coronary arteries resistance recovery to the base level in reperfusion period was more in these animals than in healthy dogs. In severe alloxane diabetes (hyperglycemia more than 12 mmol/l), the reflectory components of circulation reactions during myocardial ischemia, namely heart contractility function decrease, bradycardia, peripheral vessels resistance and arterial blood pressure reduction, were weakened or even absent, but the recovery velocity of cardiohaemodynamic parameters and the level of metabolic processes in myocardium was significantly slowed in the reperfusion period. PMID- 9221123 TI - [Intercellular interactions during the healing of an experimental skin wound]. AB - On the model of skin wound in white rats in the dynamics of wound process the interaction of mast cells and fibroblasts was studied. Quantitative, morphological and functional changes of mast cells, the content of free and total histamine and serotonin, the reaction of fibroblasts in the focus of skin lesion during 20 days after injury were investigated. It was established that the mast cells played an important role not only in the early inflammatory phase but also in the later time of wound process. An increase in the mast cells count, in their functional activity, in free and total histamine and serotonin content during the reparative phase of wound process corresponding to the fibroblast reaction was observed. The increase in the mast cells number, the greatest release and accumulation of biogenic amines were paralleled by simultaneous increase in the fibroblast count, enhanced forming of new connective tissue, by drastic reduction of the area wound. The possible mechanisms of mast cells and fibroblasts interaction in the reparative phase of wound process are discussed. PMID- 9221124 TI - [Mast cells in a focus of carrageenan-induced acute aseptic inflammation]. AB - On the model of carrageenen-induced acute aseptic peritonitis in rats the regularities of the functional state of mast cells (MC), of the content of free and cellular histamine in exudate and mesenterium and histamine in blood from the 5th minute up to 10th day after injection of phlogogene have been studied. Also as in infectious inflammation, the phasic reaction of MC which is observed at least during 10 days after action of phlogogene has been shown. The quick short first phase during half an hour after induction of inflammation has been observed and the gradual long second phase from the 1st hour up to 10th day has been noted. At least during 1st day the regularities of reaction of MC were extremely similar to the ones in infection peritonitis. Also as in infectious inflammation the second phase of reaction of MC correlates with the accumulation of leukocytes in inflammatory focus. At the same time the dependence of dynamics of functional changes of MC in aseptic inflammation on the properties of phlogogene (irritating or relatively indifferent phlogenes) has been established. PMID- 9221125 TI - [The effect of complete fasting on the structural-functional organization of the white pulp of the spleen]. AB - The work presents the results of morphologic, electron microscopic and morphometric assessment of the liver in white mongrel rats in the dynamics of complete long fasting. The investigations reveal that 7 days complete fasting in animals is accompanied by a certain dynamics of structural-functional reconstruction in various zones of splenic white pulp. The periarterial area shows microlymphocytes number decrease (T-dependent zone). However, B-dependent area of splenic white pulp experiences more significant reducing lymphocytes. Via electron microscopic observation pit-cells ultrastructure (the tissue form of large containing granules lymphocytes possessing a natural killing activity) has been ascertained. It is of note that in the dynamics of complete fasting there occurs remarkable increasing a relative pit-cells quantity, these pit-cells containing osmophilic macrogranules, proving their sharp functional activity rise. Complete long fasting demonstrates sufficient immunosuppressive effect with simultaneous enhancing nonspecific resistance of the organism due to a greater amount of natural killers and ascending their functional activity. PMID- 9221126 TI - [The clinico-hemodynamic and psychological characteristics of the course of hypertension in those who cleaned up the aftermath of the accident at the Chernobyl Atomic Electric Power Station]. AB - A comparative evaluation of 53 liquidators of the Chernobyl NPP accident and 49 subjects who were not exposed to ionizing radiation, patients with stage II hypertensive disease (HD), revealed clinical features of the disease course, apparent changes in the psychological profile and vegetative supply of the reactions of the cardiovascular system in the former category of the patients. Based on the reactions to the functional tests and urinary excretion of catecholamines, low threshold was shown of resistance to psychoemotional stress in the liquidators, as was usefulness of changes in alpha- and beta adrenoblockers. PMID- 9221128 TI - [Contralateral pneumothorax occurring during cardiopulmonary resuscitation]. AB - The author describes a case of contralateral pneumothorax and speculates upon difficulties inherent in its diagnosis to be made with no delay. In conclusion, the author emphasizes the importance of diagnosing, in a timely fashion, of traumatic (spontaneous?) pneumothorax which may contribute to failure of cardiopulmonary resuscitation. PMID- 9221127 TI - [The treatment of Hodgkin's disease by taking into account individual sensitivity to antitumor preparations]. AB - Polychemotherapy as one of the chief modes of treatment of Hodgkin's disease needs to be updated time and time again. Among approaches toward optimization of chemotherapeutic effects of the drug preparations in question is their selection tailored to the individual with regard to the results of the preliminary investigation into sensitivity to cytostatics. At the department of systemic tumor diseases, treatments with regard to individual sensitivity to chemotherapeutic drug preparations were administered to 190 patients with lymphogranulomatosis. The above treatments appeared to be associated with significant improvement of the immediate results, in the patients developing drug resistance included. PMID- 9221130 TI - [Citation rates in the assessment of the scientific output of medical scientists as a model study method in scientometry]. PMID- 9221129 TI - [Polyserositis in a female patient with hypothyroidism]. AB - A case is described of hypothyrosis, with an exudate coming into the serous cavities (pericardium, pleura, peritoneum) being a predominant symptom in the clinical picture of the condition. It took three months for the events related to polyserositis to dispel completely as a result of the substitution hormonotherapy. There were no sings of decomposition of hypothyrosis (recurrence of exudation in the serous cavities) during a 6-year prophylactic management. The authors maintain that polyserositis is suggestive of hypothyrosis, so can be of help in the diagnosis of this medical condition. PMID- 9221131 TI - [The concept of a scientific and practical approach to research into the mechanism of the situational type of management in the public health system]. PMID- 9221132 TI - [Is radioiodine ablation of residual thyroid tissue indicated after an operation for differentiated thyroid cancer?]. PMID- 9221133 TI - [The mechanisms of action and clinical importance of cholinergic blockers]. PMID- 9221135 TI - [Nondrug methods of treating hypertension]. PMID- 9221134 TI - [The clinico-hematological characteristics of the participants in the cleanup of the aftermath of the accident at the Chernobyl Atomic Electric Station with stable deviations in blood analyses]. AB - On a screening examination of 6145 individuals who took part in the elimination of Chernobyl breakdown and were exposed to radiation in the range of 0.05-1 Gr, a group was identified of those subjects displaying stable deviations from the norm in their blood analyses, such as leukocytosis, leukopenia, thrombocytopenia, anemia, lymphocytosis, monocytosis, and some other abnormalities. A 3-yr follow up revealed 17 cases of malignant diseases of the hematopoietic system, suggesting high risk for hemoblastoses morbidity. Comparative characterization is given of hematologic deviations in blood analyses together with the clinical status of persons who took part in the elimination of the effects of ChNPP accident in 1986 and those subjects working permanently within a 30-km radius. PMID- 9221136 TI - [The clinical use of rifathyroin]. PMID- 9221137 TI - [The pathomorphological aspects of drug abuse and addiction]. PMID- 9221138 TI - [The medical ecological consequences of the Chernobyl catastrophe]. PMID- 9221139 TI - [Immune and fibrinolytic system functions in the cardiac manifestations of atherosclerosis]. AB - A total of 77 patients with various clinical forms of cardiac atherosclerosis were examined. Fibrinolytic and immune systems were studied together with blood plasma concentration of fibronectine. Disorders of the same type have been revealed of immunologic reactivity manifested by enhancement of humoral immunity and impairment of cellular immunity, increase in coagulative activity, decline in the activity of the system of fibrinolysis. Latent DBS syndrome recorded in the above patients associated with decline in the activity of monocytic-macrophagal link of immunity and low fibrinolytic potential is a pathogenetic prerequisite of progression of atherosclerosis indicating main trends in drug therapy. PMID- 9221140 TI - [Viral infection and rheumatic diseases]. AB - 13 patients with generalized hyperergic vasculitides and 20 patients with rheumatism were examined for antibodies against antigens of viruses of hepatitis B and C. In the majority of patients with vasculitides and almost half of those with rheumatism, there have been revealed antibodies against antigens of hepatitis B or C. These findings allow some judgement about importance of viral infection in the development of autoimmune and infectious and allergic diseases, which observation may be related to a significant depression of the suppressor function of T-lymphocytes by viral infection. PMID- 9221141 TI - [The role of the killer activity of the peripheral blood lymphocytes in the pathogenesis of rheumatism]. AB - A total of 49 patients with rheumatism running a sluggish course, were examined, as were those with decompensated tonsillitis, systemic lupus erythematosus, and 20 essentially healthy subjects. In inactive rheumatism an increase in the killer activity of lymphocytes and augmentation of amounts of Fc+lymphocytes were revealed, which values were significantly different from those in the other groups. The detected changes in the activity of lymphocytes may serve as differential-diagnostic criteria. PMID- 9221142 TI - [The interdigestive motility of the gallbladder in patients with gastric and duodenal peptic ulcer]. AB - As many as 30 patients with uncomplicated ulcer disease and 8 essentially healthy individuals were studied for the tone and contractile function of the gallbladder under basal conditions with regard to the phases of duodenal interdigestive motility. In normals, the gallbladder functions according to phases of duodenal interdigestive motility while the clinical picture in those patients with ulcer disease is distinguished by predominance of hypermotor dyskinesia of the gallbladder and bile tract, which fact is manifested by reduction of its capacity as well as hypertone and discordant work of Lutken's--and Oddi sphincters leading to untimely and incomplete outgo of the bladder bile. Ultrasonic investigation of the gallbladder is to be carried out with due regard to its phase-associated interdigestive motility. PMID- 9221143 TI - [The clinical aspects of the pathomorphosis of tuberculosis of the respiratory organs in adults (based on data from autopsy material)]. AB - With the purpose of studying the clinical aspects of pathomorphosis of tuberculosis of the respiratory organs of different genesis in adults, a comparison was done between the clinical data and results of postmortem examinations of necropsies of 1814 those declased who had died of primary (n = 250) and secondary (n = 1564) tuberculosis over 45 years (1947-1992). Important changes were found out to occur in the clinical course of primary and secondary tuberculosis of the respiratory organs in adults secondary to induced pathomorphosis. Deaths of primary forms of tuberculosis had reduced from 20.3% to 5.4%, while those of secondary tuberculosis increased from 79.7% to 94.6%. In tuberculosis of primary genesis, major proportion of the patients died as a result of the development of generalized forms of the specific process and marked tubintoxication, in secondary tuberculosis they died from formation of chronic cor pulmonale and cardiopulmonary insufficiency. At present, reversion is identified of grave forms of primary and secondary tuberculosis which were common in the past. PMID- 9221144 TI - [The dynamic surface tension of the blood and urine in chronic glomerulonephritis]. AB - Dynamic surface-tension (S-T) was investigated of blood and urine of healthy individuals and patients with chronic glomerulonephritis by the proposed method of maximum pressure in a bubble with the aid of the computer-assisted device tensometer MPT-1 (Lauda, Germany). Patterns of changes in dynamic and static S-T were found out as were those of the slope of the curve in patients with mesangioproliferative and mesangiocapillary variants of the condition, in nephrotic syndrome and chronic renal insufficiency. Correlation type comparison was performed of physical-and-chemical properties of biological fluids to parameters characterizing protein and fat exchange. Relationship between dynamic S-T and biochemical composition of blood and urine was revealed. Diagnostic value of results of the studies made is discussed. PMID- 9221145 TI - [The pathogenesis of infertility in epididymitis]. PMID- 9221146 TI - [The characteristics of the psychoautonomic status of patients with chronic combined diseases of the digestive organs]. AB - Studied with the aid of the variational pulsimetry, adapted Luscher's [correction of Leucher's] colour test, were the functional state of the vegetative nervous system, vegetative homeostasis, psycho-emotional state during both exacerbation and clinical remission of associated digestive diseases. Overall forty six children aged 11-14 were examined. The children were found out to be unlike in respect of the vegetative homeostasis. A contingent was identified with their compensatory and adaptive mechanisms failing to work adequately, and showing inability to react in an operative manner, high level of anxiety, emotional stress, psychic fatigue. Note is taken of the fact that in clinical remission the above parameters fail to return to normal whereupon an approach tailored to the individual is warranted toward treatment of the primary disorder to prevent relapse and chronization of the morbid process. PMID- 9221147 TI - [The use of laser reflexotherapy in the combined treatment of patients with atherosclerotic circulatory encephalopathy]. PMID- 9221148 TI - [Microhemodynamics and energy metabolism in schizophrenia patients]. AB - An apparent disturbance was revealed in microhaemodynamics of patients diagnosed as having schizophrenia (n = 210) which was more pronounced in continuously progredient form of the above medical condition. An increase in conjunctival indexes, polymorphic character of capillaries, decrease in numbers of capillary loops were revealed by biomicroscopy of the bulbar conjunctiva and capillaroscopy respectively. The patients showed lowering of ATP level and rise in the content of cathodic LDG4-LDG5 fractions, accumulation in blood of lactic and pyruvic acids. PMID- 9221149 TI - [The direct and reverse processes of cholesterol and triglyceride transport in patients with Alzheimer's disease]. AB - Based on the investigation of 42 persons with Alzheimer's disease, it has been shown that in spite of absence of changes in the sum of apoB-containing fractions of lipoproteins a redistribution of their particles is observed suggesting adverse shifts in the processes of direct transport of lipids. These patients also manifest disturbances of the reverse transport of lipids which, fact is inferred from a perverted redistribution of particles of high-density lipoproteins. PMID- 9221150 TI - [A methodological approach to determining electrolyte disorders in cholera patients]. AB - A total of 198 cholera patients were studied for blood concentrations of electrolytes; the above patients were treated at Mykolaiv Cholera Hospital during an outbreak in 1995. It is advisable that blood plasma concentration of electrolytes be represented as mmol/kg of the mass of a cholera patient's body instead of mmol/l, to indicate disturbances in electrolytic balance. It is a matter of principle for the assessment of the patient's state to be done, first of all, before initiating the rehydration therapy treatments. Determinants of electrolytes in cholera patients got decreased not only in severe course of the illness but also in moderately severe one. Of all the electrolytes studied in blood plasma, it is in K+ and Cl- that deviations from the norm were at their greatest. Since electrolytic balance is a sensitive indicator of homeostasis of human organism it is useful to calculate the volume of salt solutions for the primary rehydration according to blood plasma concentration of Cl- in mmol/kg of mass of the patient's body. Lowering of electrolyte concentration in erythrocytes occurred only in severe course of cholera involving K+ only. PMID- 9221152 TI - [Diagnostic criteria for systemic scleroderma]. AB - Diagnostic informative value was studied of a set of N. G. Guseva's diagnostic criteria (1993) for systemic sclerosis (scleroderma). A total of 104 patients with systemic sclerosis (scleroderma) and 126 patients with other rheumatic diseases were examined. The above set of criteria was found out to be of high sensitivity and specificity--99% and 98.4% respectively. The authors present their own modification of the set of diagnostic criteria by N. G. Guseva (1993) which is not inferior in respect of sensitivity and specificity to the original but is more simple, and with a greater potential for further applications, due to which features it can come into widespread use. PMID- 9221151 TI - [The possibility of using highly disperse iron for the directed transport of thyroxin in the body]. AB - It has been ascertained that high-disperse powders of iron obtained by a thermochemical method in the process of formation of particles with pre determined physical-and-chemical and medical-and-biological properties (essentially monodisperse, harmless, capable of exerting a bactericidal action, having an as-needed specific surface, corrosion resistance, withstanding sterilization heat up to 120 degrees C, allowing the magnetic characteristics to be controlled, etc.) can be used for directed transport of medicinal substances (thyroxin), hormone control, protection of the organism from overdosing of the administered drug preparations. Feasibility is shown of prolonged administration of thyroid hormones through the gastrointestinal wall with the aid of high disperse ferromagnetic compositions as containers-carriers, which fact may secure the organism-controlled maintenance of the thyroid status in various thyroidal pathologies. PMID- 9221153 TI - [The intraoperative diagnosis of functional disorders of the major duodenal papilla in patients with acute biliary pancreatitis]. PMID- 9221154 TI - [Body nonspecific resistance in patients with gastric and duodenal peptic ulcer]. AB - The present study focuses on bodily resistance to Helicobacter pylori infection. A decline is shown in factors of unspecific resistance, in particular, leucocyte content of non-enzymic cationic proteins in the phase of exacerbation of the ulcer process. Weakening of resistance is accompanied by a decline in the activity of phagocytosis. Thus, compromised antibacterial defence of the organism during the phase of exacerbation of the ulcer process contributes to the development of Helicobacter pylori in the gastroduodenal zone. PMID- 9221155 TI - [The efficacy of using prolonged-action L-asparaginase in a treatment program for acute lymphoblastic leukemia in adults]. PMID- 9221156 TI - [The use of thienam for the emergency antibacterial therapy of acute suppurative pyelonephritis]. AB - Results of a clinical and microbiological investigation designed to study thienam permit recommending this drug as an urgent agent of choice in acute purulent pyelonephritis. The drug is well tolerated. Among other benefits is extremely high sensitivity of bacteria (pathogenic and conditionally pathogenic) to the medication, achievement of an expected clinical effect within a short time since its therapy institution. PMID- 9221157 TI - [The clinico-immunological effects of leukinferon in the treatment of patients with acute suppurative pyelonephritis]. AB - A many as 20 patients with acute purulent pyelonephritis were studied for clinical and immunological effects of leukinferone. The above patients were convalescent earlier than control subjects. There were instances where clinical status was not accompanied by shifts in immunity. However, on an average in the group, such parameters as level of lymphocytes, amounts of B-cells, Igs G, A, T helpers, and Tx/Tc ratio were found to be increased as was activity of EK-cells. Stimulating effects of leukinferone were particularly pronounced in those patients with decreased values for immunological status. A conclusion is reached to the effect that patients with acute pyelonephritis may benefit from those treatments involving the use of the above drug preparation. PMID- 9221158 TI - [The effect of Moradol (butorphanol tartrate) on immunity]. AB - Two groups (with n = 5 in each group) were studied of essentially healthy volunteers by double-blind method. Those subjects in group 1 were administered placebo, those in group 2 received 0.1 mg/kg of moradole. In both groups, blood to be analysed was taken in fixed time intervals. An investigation designed to study immunological parameters involved correlation of results obtained before and after administration of moradole or placebo. Opioid agonists are endowed with marked immunity-inhibiting effect disappearing after the administration of naloxone. A hypothesis has been formed to the effect that those drug preparations endowed with agonist-antagonist activity toward opioid receptors fail to induce immunodepression, otherwise it is less pronounced than that of opioid agonists. The studies made allow a conclusion to be reached that administration of moradole is not associated with immunodepression. PMID- 9221159 TI - [Screening for malignant neoplasms by using the new method of marker diagnosis, the Oncotest]. AB - The proposed method for tumor screening ("Oncotest") is based on detecting substances (calcium-histone complexes) in the human blood which are markers of malignant neoplasia. Using "Oncotest" in a series of 5411 cases, we identified malignant tumors of various histogenesis (cancer, sarcoma) arising in different sites in 69 cases. "Oncotest" is especially important for early (pre-clinical) detection of malignant neoplasms during preventive population screenings. PMID- 9221160 TI - Ernest Hemmingway and Sir William Osler. PMID- 9221161 TI - Asthma among the famous. Brazilian literary figures. Augusto dos Anjos (1884 1914). PMID- 9221162 TI - Asthma among the famous. Brazilian literary figures. Graciliano Ramos (1892 1953). PMID- 9221163 TI - Asthma among the famous. Brazilian literary figures. Otto Lara Resende (1922 1992). PMID- 9221164 TI - Asthma among the famous. Antonio Guzman Blanco (1829-1899). Eleventh president of Venezuela. PMID- 9221165 TI - Asthma among the famous. Juan Vicente Gomez (1857-1935). Eighteenth president of Venezuela. PMID- 9221166 TI - Asthma among the famous. Gabriel Turbay (1901-1948). Columbian physician and diplomat. PMID- 9221167 TI - Asthma among the famous. Ernesto "Che" Guevara (1928-1967). Argentinean physician and revolutionary. PMID- 9221168 TI - New dietary strategies for the prevention and management of cardiovascular disease. Proceedings. London, Ontario, Canada, June 20, 1996. PMID- 9221169 TI - The future of the global physicians' movement. PMID- 9221170 TI - [Congenital duodenal diaphragm in an adult associated with an abnormal location of Vater's ampulla: a case report and review of the literature]. AB - Web-related duodenal obstruction in the adult is rare, so is the anomalous location of the ampulla of Vater in the third part of the duodenum. A woman who harboured both anomalies is described and the literature reviewed. The diagnosis, often made at laparotomy, mandates an exploratory duodenotomy. The best treatment consists of partial excision of the web and transverse closure of the duodenum. A duodenojejunostomy or duodenoduodenostomy, or both, are acceptable therapies. The anomalous location of the ampulla of Vater in the third part of the duodenum does not require treatment. PMID- 9221172 TI - [Current activity of the World Health Organization in the area of systematic development of quality in health care]. AB - The European Regional Office of WHO develops in recent years overall activities to promote continuous quality development in member countries. Several innovated targets and indicators of the strategy of the programme Health for All as well as the extensive definition apparatus elaborated by the department for ensuring the quality and evaluation of technologies is also focused on improving the quality of care. In the submitted article the author discusses the basic philosophy of procedures of the systematic development of the quality of care which resulted from the common effort of the European Regional Office of WHO and the Association of National Medical Societies. The European Regional Office in collaboration with some European health institutions and medical societies developed also information systems and models of systematic development of quality of care which are excellent examples of what can be achieved on a horizontal level. This applies in particular to the programmes DIABCARE, WHOCARE, ORATEL and OBSUOUID. In the submitted paper examples are given of the implementation of these programmes as well as the pertinent fundamental references. The author mentions briefly also some forms of implementation in some advanced European countries. In the conclusion the author deals briefly with the essential aspect of quality of health care, i.e. the relationship of effectiveness and economy of health care. PMID- 9221171 TI - Quiz case of the month. Multilocular cystic nephroma (MLCN). PMID- 9221173 TI - [Somatosensory evoked potentials in panic disorders]. AB - BACKGROUND: Panic disorders are at present the most intensely studied psychiatric entity calling for a multidisciplinary approach. After promising results following the use of evoked auditory potentials for the first time in panic disorders the somatosensory evoked potential was tested. METHODS AND RESULTS: Fifteen patients with panic disorders or agoraphobias or without the latter (12 women, 3 men, average age 33.7 +/- 9.3 years) were subjected during the interparoxysmal period to examination of the somatosensory evoked potential (SEP) of the n. medianus, bilaterally. The control group was formed by 20 healthy subjects (12 women and 8 men), age 20-45 years, average age 34 +/- 8.5 years). An EMG apparatus Madaus Electronic 15 was used. The authors revealed significant changes of some parameters of SEP only in patients with panic disorders: bilateral shortening of the mean latency P17 (P < 0.01). Bilateral shortening of the mean conduction time N13-P17 and P14-P17 (P < 0.01). CONCLUSIONS: The assembled findings are consistent with the pathophysiological concept of panic disorder at the brain stem level. The findings are confronted with changes of auditory evoked potentials and are consistent with contemporary views as regards pathomechanisms of panic disorders. PMID- 9221174 TI - [Frequency of cytotoxic and helper T-lymphocyte precursors in donors with alloreactivity to recipient histocompatibility antigens. Possible use in predicting acute graft vs host disease in bone marrow transplantation]. AB - BACKGROUND: The utility of cytotoxic T lymphocyte precursor (CTLp) and helper T lymphocyte precursor (HTLp) frequencies estimation for detecting alloreactivity and for the prediction of acute graft versus host disease (aGVHD) has been evaluated. METHODS AND RESULTS: The limiting dilution assay and a maximum likelihood statistical programme were used for CTLp and HTLp frequency estimation. A high CTLp frequency suggesting the presence of hidden class I mismatches was detected in 41.2% of unrelated pairs. HLA-A and -B matched by serological typing and DRB1 and DQB1 matched by DNA analysis. Severe aGVHD (grade III-IV) occurred in all patients of this group who underwent bone marrow transplantation (BMT). In two patients of the three evaluated with low pretransplant CTLp frequency a mild form (grade I) or no aGVHD developed after unrelated BMT. Positive frequency of alloreactive HTLp was found in 50% of HLA matched unrelated pairs. The comparison of CTLp and HTLp values in the same individuals showed that these two methods are not fully alternative in detecting alloreactivity. In the group of HLA identical siblings, 18.7% of positive HTLp results were only found. Besides HLA-DP incompatibilities, the differences in non HLA genes could cause this alloreactivity. CONCLUSIONS: CTLp assay has a potential for the prediction for aGVHD development following BMT from HLA matched unrelated donors. CTLp results suggest the necessity of more accurate class I typing in these cases. The comparison of CTLp and HTLp frequencies showed that the results can differ in some unrelated donor-recipient BMT pairs suggesting the convenience of simultaneous performing of both assays for the alloreactivity assessment. More cases have to be considered to determine the relationship between pretransplant HTLp frequency and posttransplant aGVHD development in HLA identical siblings. PMID- 9221175 TI - [Selection and long-term effect of treatment in vasovagal neurocardiogenic syncope]. AB - BACKGROUND: Syncope state performed by transient loss of consciousness connected with a postural tone decrease present still an important therapeutical problem. The authors use HUT testing for discovering of vasovagal etiology of a syncope. The aim of the study was to find out the optimal medication in treatment and prevention of VVS by repeated HUT drug testing in a group of positive patients. METHODS AND RESULTS: We examined a group of 300 patients (172 female and 128 male) age 35.26 +/- 13.47 years with an anamnesis of recurrent syncopes by HUT test. In 83 patients (27%) we set up a diagnosis of VVS. Thirteen patients (15.66%) were only followed without medication. The rest of the group (70 patients, 84.34%) started with a therapy. Testing was performed by repeated HUT according to the "Westminster protocol". HF and BP were measured by FINAPRESS 2,300 made by Ohmeda. The effective therapy was found in 64 patients (91.43%) of the group. For various reasons the therapy was changed and retested in 9 patients (14.06%). Beta1 selective beta-blockers were used in 51 patients (80.96%) theophyllin in 5 patients (7.94%), etilephrin in 6 patients (9.52%) and clonidine in 1 patient (1.59%). CONCLUSIONS: The therapy of VVS should be set-on according to the effective drug selection by serial drug testing in repeated HUT tests. The article shows a high predictive value of negative HUT test for a long term effect of tested therapy. PMID- 9221176 TI - [Use of CA repeat polymorphism in direct and indirect diagnosis of Duchenne and Becker muscular dystrophy]. AB - BACKGROUND: DNA analysis makes it possible to confirm the clinical diagnosis of the majority of cases DMD/BMD and to detect at the same time carries and the prenatal diagnosis for relatives at risk. The objective of the present work was to improve the haplotype analysis and to identify the most frequent deletions in carries. METHODS AND RESULTS: The method is based on the initial amplification of several DNA polymorphisms of CA repetitions and subsequent identification of alleles in the denaturation sequencing polyacrylamide gel using radioactive detection system. The system of CA polymorphisms provides information in the great majority of families, it detects the recombination in the DMD gene, which reduces to a minimum the risk of a diagnostic error and provides valuable information on the carriership of deletion. CONCLUSIONS: The introduction of haplotype analysis of CA repetitions is beyond doubt an asset to the prenatal diagnosis of DMD/BMD and assessment of carriership of this serious hereditary disease. The variability of the length of alleles of these markers improves analysis, prenatal diagnosis and makes it possible to rule out or identify deletion in cca 40% carriers. PMID- 9221178 TI - [Failures, problems and hopes in the transformation of health services. I. Causes of erroneous decisions in the beginning phases of transformation]. AB - It is generally accepted that the transformation of our health services was not a success. The reasons of failure were political and professional. Our country was too long isolated from the West and we lacked experience of countries where the reform of health services had been underway for some time. At the onset of the reform three basic findings were underrated or omitted: 1. Health needs are infinite and the resources of the health services have limits which cannot be trespassed. 2. There is no simple linear correlation between financial expenditure and the health standard. 3. The main problems of health care cannot be mastered by mere deetatization, privatization and a free market. PMID- 9221177 TI - [Enzyme therapy in children with severe forms of Gaucher's disease]. AB - The enzyme therapy with Ceredase in patients with Gaucher's disease is at present probably the most expensive treatment in the whole world. One-year treatment of an adult patient with Gaucher's disease costs more than 7 million crowns. Indications for treatment in individual patients as well as financial provisions are so far problematic in the Czech Republic. From a total of 28 patients of varying age with Gaucher's disease diagnosed by the authors Ceredase was administered to two boys with a severe course of the disease. Within one year of treatment the health status of both children improved, growth became normal, the spleen diminished in size by 20-35%, haematological manifestations of hypersplenism are receding, there was a 32-46% decline of the activity of serum chitotriosidase and biochemical parameters of the disease improved. PMID- 9221179 TI - [Hepatobiliary complications of idiopathic intestinal inflammations]. AB - Extraintestinal manifestations and metabolic complications are very frequent in patients with idiopathic inflammations of the gut and are encountered in at least 35% of these patients. In Crohn's disease extraintestinal manifestations are more frequent than in ulcerative colitis, in particular when the large bowel is affected. Metabolic complications are the result of inflammatory changes of the small intestine or develop as a result of the reduced reabsorption surface of the gut. As to the relationship to the activity of the idiopathic inflammation of the gut, extraintestinal manifestations can be differentiated into those which depend on the activity of the basic disease and those which lack this dependence. From the aspect of a long-term prognosis extraintestinal manifestation independent on the activity of the inflammation of the gut are much more serious, because as a rule they have a long-term and usually progressive trend. The most serious extraintestinal complication is primary sclerotizing cholangitis which in the majority of patients leads to destruction of the biliary pathways and the development of biliary cirrhosis. Depending on the predominantly affected site of the biliary system, primary sclerotizing cholangitis is divided into three types. It is encountered much more frequently in ulcerative colitis than in Crohn's disease. Treatment of primary sclerotizing cholangitis is not very effective. At present it appears that the only drug with an effect on the course of the disease is long-term administration of urodesoxycholic acid. For patients with manifestations of hepatic insufficiency the only solution is transplantation of the liver. In all patients where the diagnosis of primary sclerotizing cholangitis was established, at the same time the possibility of inclusion in a transplantation programme should be considered. The relationship between sclerotizing cholangitis and pericholangitis has not been resolved conclusively. At present the majority of authors is inclined to believe that pericholangitis is part of changes associated with sclerotizing cholangitis. Other hepatobiliary complications of idiopathic inflammations of the gut such as cholelithiasis and parenchymatous liver damage, steatosis of the liver and chronic autoimmune hepatitis are not such a serious problem as sclerotizing cholangitis. PMID- 9221180 TI - [The effect of disability of the aged patient on the level of caregiving burden by the family]. AB - BACKGROUND: A growing number of dependent elderly people is cared for at home by family members. However, long-term caregiving may become an intolerable strain for some families and lead to failure of family care. The aim of the study was to examine if level of physical and mental disability of the patient influences the extent of perceived caregiver burden representing risk factor for negative outcome. METHODS AND RESULTS: 128 elderly patients with disability and dependency (37 men, 91 women, average age 79.9 +/- 6.9 yrs) and 128 their primary caregivers, mostly family members (28.9% men, 71.1% women) were evaluated. Functional status of care recipient was assessed by means of Barthel ADL Index (mean = 70.9 +/- 26.5), IADL Test (mean = 31.4 +/- 23.5) and Mini-Mental State Exam, MMSE (mean = 20.4 +/- 6.5). Average score of Caregiver Burden Interview (CBI) was 34.7 +/- 18.8. According to CBI, 40.6% of caregivers were found under high or even extremely hig level of stress. Level of perceived burden correlated significantly with physical and mental disability level, in decreasing order for IADL, ADL and MMSE (rs = 0.582-0.708, p < 0.001). CONCLUSIONS: Caregiver burden of family caregivers is significantly related to the level of functioning and cognitive impairment of care recipient, particularly to his/her ability to perform instrumental activities. Functional decline of elderly patient represents a risk factor which contributes to negative caregiving outcome and institutional placement. PMID- 9221181 TI - [Familial hypobetalipoproteinemia]. AB - BACKGROUND: Familial hypobetalipoproteinaemia (FHBL) is a relatively rare inborn error of metabolism which must be considered in the differential diagnosis of hypocholesterolaemia which cannot be explained by secondary causes (severe malnutrition, generalization of neoplastic disease etc.). METHODS AND RESULTS: In the submitted paper the authors present the results assembled in a family with four heterozygotes with FHBL. The proband is a 27-year-old woman (total cholesterol (TCh) 1.70, triacylglycerols (TG) 0.20, HDL-cholesterol (HDL-ch.) 1.38, LDL-cholesterol (LDL-ch.) 0.34 all in mmol/l, apolipoprotein B (apo B) 0.25, lipoprotein(a)(Lp(a) 0.09 all in g/l, isoforms of apolipoprotein E/E3 (iso apo E). Mother (age 53 years) of the proband (TCh 3.06, TG 0.37, HDL-ch. 1.99, LDL-ch. 0.90 all in mmol/l, Apo B 0.37, Lp(a) 0.14 all in g/l, iso apo E3/E3)). Two of the proband's sisters (23 and 20 years) (TCh 3.92 and 2.55 resp., TG 0.57 and 0.23 resp. HDL-ch. 1.86 and 1.63 resp., LDL-ch. 1.80 and 0.82 resp. all in mmol/l, Apo B 0.73 and 0.37 resp., Lp(a) 0.47 and 0.63 resp. all in g/l, iso apo E2/E3 and E2/E3 resp.). The diagnosis confirms the autosomal dominant transmission. During the proband's pregnancy (during the 38th week), contrary to normocholesterolaemic women the TCh did not rise (1.43 mmol/l, the TG level (0.62 mmol/l), Apo B (0.43 and Lp(a) 0.18 all in g/l rose. CONCLUSIONS: According to the available literature this is the first description of FHBL in our literature and a priority investigation of plasma lipid concentrations, lipoproteins and apolipoproteins in a women with a heterozygous form of FHBL during pregnancy. PMID- 9221182 TI - [A controlled clinical trial of Consupren versus cyclophosphamide in chronic glomerulonephritis]. AB - BACKGROUND: Experience gained from recent studies shows, that Cyclosporine-A (Cy A) may decrease proteinuria (PU) in some forms of chronic glomerulonephritis (GN) with the nephrotic syndrome. The aim of this study was to test the efficacy of Czech-made Cy-A, Consupren. METHODS AND RESULTS: 30 patients with chronic GN, confirmed by biopsy and PU higher than 3 g/d, corticodependent or corticoresistant, were randomized according to the month of birth to either therapy with Consupren at an initial dose of 5 mg/kg/d (CS group, after dropout of 3 patients who did not finish the treatment, n = 16) or Cyclophosphamide at a dose of 1.5 mg/kg/d (K group, n = 11), and prednisone maintained at the original dose in both groups. The treatment was stopped after six months or after achieving remission. The main criterion of efficacy was PU. The decrease in mean values, statistically evaluated by Holm's procedure was highly significant in the CS group and non-significant in the K group. A similar evaluation of PU corrected by glomerular filtration rate was significant in both groups. Partial or complete remission was reached in 50% of CS group patients and in 34% of K group patients (NS). In the CS group a significant increase in the mean values of albumin and gama-globulin, and a decrease in cholesterol levels were observed. In the K group, these changes were non-significant. CONCLUSIONS: In patients with chronic GN and the nephrotic syndrome, the efficacy of Consupren treatment gives comparable, or even better results versus treatment with Cyclophosphamide. PMID- 9221183 TI - [The effect of parasympatholytics on the therapeutic effectiveness of the oxime HI-6 against organophosphorus compounds (Soman, substance VX, Fosdrin) in mice]. AB - BACKGROUND: Causal antidotal therapy of acute intoxications with organophosphorus compounds involving administration of the parasympatholytic and cholineesterase reactivator (oxime) has not been resolved so far satisfactorily despite knowledge of the basic mechanism of action of these noxious substances. METHODS AND RESULTS: In experiments on mice the therapeutic effect of parasympatholytics atropine, benactyzine and biperidene (Akineton) combined with oxime HI-6 on the toxicity of highly toxic organophosphates soman and substance VX and the organophosphorus insecticide phosdrine was compared as regards their influence on the LD50 of these noxious substances during 24-hour survival of experimental animals. Two levels of antidotes were tested. These findings confirm that the LD50 value of untreated intoxication with all three organophosphorus compounds is most increased by oxime HI-6 combined with benactyzine regardless of the antidote dosage. CONCLUSIONS: Oxime HI-6 is the most effective against highly toxic organophosphates and organophosphorus insecticides when combined with the centrally acting parasympatholytic benactyzine. PMID- 9221184 TI - [Failures, problems and hopes in the transformation of health services. 2. Dilemmas in the development of health services]. AB - In the second stage of the reform it is essential to proceed from problems of financing and rewards to health services and their ethic attributes. The dilemma produced by the increasing demands on availability and standard of services on the one hand and efforts to reduce costs on the other leads to the metaphorical "iron triangle of health care", exposed to intense internal tension. The only possibility how to straighten it, which would weaken the danger of crisis, are strict regulatory measures enforced by the State. The foremost task is to define and specify basic (standard) care. As a possible approach the author mentions the Oregon experiment. He draws attention to the phenomenon of "concentration of costs" in the practice of health insurance companies, calling for a major measure of solidarity. PMID- 9221185 TI - [The hibernating myocardium--its use in clinical practice]. AB - Hibernated myocardium is a term used to describe a state of myocardial hypocontractility which is due to chronic hypoperfusion of the heart muscle. The myocardium remains, however, viable and therefore revascularization can significantly improve the reduced mechanical function of the left ventricle. Differentiation between hibernated myocardium and a myocardium which was irreversibly damaged and replaced by scar tissue is particularly important in patients with ischemic dysfunction of the left ventricle. Information on the viability of dysfunctional areas of the myocardium helps in reflections on the indication and strategy of the revascularization operation. PMID- 9221186 TI - [Conversion of cyclosporin A therapy to conventional treatment with diagnostic use of aspiration biopsy in kidney transplantation]. AB - BACKGROUND: Withdrawal of cyclosporin-A from maintenance immunosuppressive therapy involves risk of rejection. The aim of the study was to reduce the risk of rejection and to evaluate the fine-needle aspiration biopsy in predicting rejection. METHODS AND RESULTS: In 41 patients 14.4 +/- 2.6 months after the first transplantation of a cadaveric graft with good and stabilized function, cyclosporine was withdrawn from triple-drug therapy while the doses of azathioprine and prednisone were increased. Prior to the change fine-needle aspiration biopsy (FNAB) was performed and methylprednisolone 500-250-250 mg administered in 3 days. FNAB was repeated after 2 weeks. 39 patients fulfilling inclusion criteria but ineligible for the switch for different reasons served as a control group. Both groups were comparable in demographic and immunological parameters. Within 3 months after conversion, rejection was observed in 3 patients (7%) vs 2 patients (5%) of the control group over a comparable period of time: and within 6 months in 6 patients (15%) and 3 patients (8%) respectively (NS). No relationship between rejections before and after conversion was found. FNAB appeared to have some predictive value for rejection. In all of the 3 patients experiencing rejection, a rejection pattern was present in the 2nd biopsy. CONCLUSIONS: Incorporation of methylprednisolone into conversion therapeutic regime decreased the risk of rejection to 7%. The rejection pattern in the second FNAB after methylprednisolone administration may be predictive for further rejection development. PMID- 9221187 TI - [Apoptosis in the morphology of Lyme borreliosis]. AB - BACKGROUND: Morphological, mainly ultrastructural changes in borreliosis has been the matter of contemporary interest indeed and it seems to be more and more clear that they are the product of an immunologic reaction. The aim of this study is to find out the epidermal changes of erythema migrans and acrodermatitis chronica on the ultrastructural level. METHODS AND RESULTS: There were 14 cases of erythema migrans and 5 acrodermatitis examined and many apoptotic keratinocytes, but also Langerhans cells and melanocytes were the dominant findings. Considering the very short time needed for apoptosis there is a good reason to take our findings as an important component of the borreliosis pathogenesis. CONCLUSIONS: The presence of many apoptotic cells in Lyme disease epidermis emphasises the importance of the immunology in the borreliosis pathogenesis. The striking number of apoptotic melanocytes suggests the higher affinity of the borrelia to the neuroectodermal tissue. PMID- 9221188 TI - [Detection of extensive deletions and duplications in the dystrophin gene]. AB - Dystrophin is a cytoskeletal protein, defects of which results in Duchenne or Becker muscular dystrophy (DMD or BMD). About 70% of all DMD and BMD cases is caused by large deletions and duplications in the dystrophin gene. Therefore, their detection at the DNA and mRNA level is the analysis of the first choice which is undergone by our patients at the molecular level. Methods which have been introduced in our laboratory for this purpose-multiplex PCR and RT-PCR-are subject of this communication. PMID- 9221189 TI - [Purging of hemopoietic progenitor cells in autologous transplantation]. AB - The authors describe the results of purification of bone marrow and peripheral progenitor cells (PBPC) for clinical transplantations. Vepeside was used to purify in 1990-1995 a total of 41 bone marrows of adults and children. Of these 23 were transplanted. Maphosphamide was used bone marrow purging in two patients; transplantation was performed in one case. By a combination of Vepeside with methylprednisolone haematopoietic cells of 24 patients were purged, transplantations were performed in 10. Three-day cultivation of haematopoietic cells in the presence of Desferal was used for purging cells of 22 patients with neuroblastoma; transplantations were performed in 10 patients. The authors give the values of nucleated cells, haematopoietic colonies of CFU-GM and CD34 positive cells obtained after purification calculated per kg body weight of the patient and the percentage yields. PMID- 9221191 TI - [Failures, problems and hopes in the transformation of health services. 3. New paradigms in health care and their necessity]. AB - Regulatory interferences of the state can only smoothen frictions in the contemporary model of health care. The new paradigm should be a fundamental breaking point as regards health care and improvement of the health status of the population. Theoretically it is based on the salutogenic concept of health promotion of WHO which shifts the centre of gravity as regards interest and activities from the sphere of disease to the sphere of health. This will involve a long-term education al process to achieve social changes of human behaviour and health culture. A major role in this social phenomenon must be played not only by health workers but in particular by civic initiatives and various public subjects such as foundations, public organizations and other interest groups and societies. PMID- 9221190 TI - [The effect of cefepime on ceftazidime, cefotaxime and imipenem resistant strains of Acinetobacter, Xanthomonas, Pseudomonas, Flavobacterium, Sphingobacterium and on producers of extended spectrum beta-lactamases (ESBL) with resistance transfer]. AB - BACKGROUND: The new cephalosporins with heterocyclic positively charged substituent at position C3 belong to the 4th generation cephalosporins. It is not evident, whereas these antibiotics are effective against cefotaxime-ceftazidime resistant bacteria. We investigated the efficiency of the cefepime (Maxipime, BMS) against Acinetobacter sp., X. maltophilia, Pseudomonas sp., Flavobacterium sp., Sphingobacterium sp. and against strains with production of ESBL (Extended Spectrum beta-Lactamases) (Enterobacter sp., Citrobacter sp., Klebsiella sp.). METHODS AND RESULTS: We determined the susceptibility of the 54 strains resistant to 50 mg/l and more of ceftazidime. The majority of the strains (35) are susceptible to cefepime and MIC is the range 1.65-3.15 mg/l of cefepime. 17 strains have MIC = 6.25 mg/l. Two strains (one strain P. aeruginosa a one strain K. pneumoniae) were resistant to cefepime and MIC = 12.5 mg/l. The activity of beta-lactamases was determined by quantitative macroiodometric method in one strain of P. cepacia and two strains of Sphingobacterium multivorum. They hydrolyse actively imipenem (they produce metallo-beta-lactamase) and ceftazidime (they are highly resistant to this antibiotic), but cefepime is more slowly hydrolysed than ceftazidime. CONCLUSIONS: At the present time cefepime is effective against ceftazidime-resistant strains in so-called "problematic" species of bacteria. However, it is important to protect its efficiency in the future, this requires its rational use. PMID- 9221192 TI - [Regulation of cholesterol metabolism with dietary addition of oyster mushrooms (Pleurotus ostreatus) in rats with hypercholesterolemia]. AB - BACKGROUND: It is generally accepted that lowering of serum cholesterol levels reduces the risk of atherosclerosis. Identification and characterization of natural substances with hypocholesterolemic activity useful in dietetic prevention or treatment of hypercholesterolemia is still relevant in countries with persistent progression of hypercholesterolemia. Addition of oyster mushroom (Pleurotus ostreatus), an industrially produced wood-rotting fungus, to the diet effectively reduced cholesterol accumulation in serum and liver of rats fed a cholesterol diet. The aim of a series of experiments was to explain the biochemical mechanism of this effect. METHODS AND RESULTS: Male Wistar rats fed a cholesterol (0.3%) diet shortly after weaning for a period of 8-10 weeks were used in the experiments. The addition of 5% of dried oyster mushroom to the diet had following effects: reduction of cholesterol level both in serum (5.12 +/- 0.55 vs. 3.44 +/- 0.16 mmol/l, p < 0.02) and liver (241 +/- 12 vs. 113 +/- 11 mmol/kg, p < 0.001); redistribution of cholesterol in favour of high-density lipoproteins; reduced production of very-low-density lipoproteins (135 +/- 7 vs. 96.5 +/- 5 mumol/h/kg, p < 0.001); reduced cholesterol absorption (61.2 +/- 2 vs. 53 +/- 2%, p < 0.02) and reduced HMG-CoA activity in liver (137 +/- 16 vs. 86 +/- 9 pmol/min/mg proteins, p < 0.02). Simultaneously, an increase in 7 alfa hydroxylase activity in liver (17 +/- 1 vs. 22 +/- 1 pmol/min/mg proteins. p < 0.02) and bile acid excretion (7 +/- 0.9 vs. 11 +/- 0.5 mg/day/rat, p < 0.02) was observed. (Values shown are means +/- SEM.) CONCLUSIONS: Biochemical mechanism of hypocholesterolemic effect of oyster mushroom on cholesterol-fed rats includes reduced production of cholesterol-rich very-low-density and low-density lipoproteins which principally determine cholesterol levels in serum. This effect is related to decreased absorption and biosynthesis of cholesterol together with increase in cholesterol catabolism and excretion of degradation products-bile acids. PMID- 9221193 TI - [The present status and future perspectives of fluorinated quinolones from the aspect of clinical microbiology]. AB - The author presents a review on the contemporary position of fluoroquinolones from the microbiological and clinical aspect. He submits basic data on the mechanism of action of these drugs and development of resistance displayed by infectious agents. Favourable pharmacokinetics make wide clinical use of these drugs possible. Indiscriminate administration could debase these drugs possible. Indiscriminate administration could debase these very valuable antimicrobial drugs. This is why the author presents in his paper the main clinical indications for their therapeutic use. To complete the problem the author makes suggestions of suitable combinations and draws attention to possible therapeutic interactions and undesirable effects, incl. contraindications. PMID- 9221194 TI - [Therapy of non-small cell lung cancer]. AB - Several regimes can provide clinical answers and longer survival to patients with small cell carcinoma of the lungs who respond to treatment. Moreover there is frequently also suppression of pathological symptoms of the disease in these patients. This situation may develop in almost 30% of patients and treatment is from the psychological aspect consistent with the patients expectations. Toxicity of chemotherapy and resulting morbidity and mortality may be followed up by strict clinical supervision and various types of supporting treatment. This supervision of patients explains probably why administration of planned treatment may be less expensive than palliative care. PMID- 9221195 TI - [Nephrotoxicity of cytostatic drugs. Pathogenesis, prevention, therapy]. AB - Tumourous diseases can damage the kidneys by different mechanisms, e.g. by infiltration of the kidneys, by obstruction of the urinary pathways or the syndrome of acute tumour lysis after aggressive cytostatic treatment. Moreover, there may be be even direct renal damage caused by cytostatic treatment. Serious nephrotoxicity with development of acute renal failure, Mg depletion and possibly chronic renal failure may be caused in particular by cisplatinum and less by its more recent derivatives. Nephrotoxicity of cisplatinum derivatives can be effectively mitigated by hydratation regimens and the administration of hypertonic NaCl. More recent findings on the nephrotoxicity of cisplatinum will certainly make possible the development of less toxic derivatives as well as more effective prevention and treatment of cisplatinum nephrotoxicity. Large-dose vaginous with iphosphamide may cause in addition to urotoxicity, common in all oxazaposphorins, the development of renal insufficiency and serious forms of acquired Fanconi's syndrome. The uro- and nephrotoxicity of iphosphamide can be reduced, but not entirely eliminated by administration of mesna. Large doses of metothrexate can precipitate in the tubules and induce acute renal failure. The nephrotoxicity of metothrexate can be prevented by alkalinisation and hydration regimes. Less common are other forms of nephrotoxicity e.g. tubulointerstitial nephritis induced by derivatives of hydroxyurea or the haemolytic-uraemic syndrome in conjunction with treatment with mitomycin C. PMID- 9221196 TI - [The Bernard-Soulier syndrome. A hereditary functional and structural disorder of blood platelets]. AB - BACKGROUND: The authors submit a clinical and laboratory description of a patient with Bernard-Soulier syndrome diagnosed in this country for the first time. The investigation comprises an analysis of haemostatic functions, selected structural indicators and the detailed morphology of thrombocytes. The analysis of defined inborn functional disorders of thrombocytes is a method for studying haemostatic mechanisms. METHODS AND RESULTS: In addition to standard haemostatic methods flow cytometry in used to assess the size of platelets, and using monoclonal antibodies against glycoproteins, their presence on the surface membrane is assessed. Thrombocytic glycoproteins are further analyzed using diagonal electrophoresis on sodium dodecyl sulphate polyacrylamide gel (SDS-PAGE). The authors found medium thrombocytopaenia with the presence of giant thrombocytes which on elecronmicroscopic examination, with the occasional exception of endoplasmic membrane proliferation, do not display structural deviations. The assessed deviation of thrombocyte aggregation after ristocetin, suggesting a defect of the adhesive capacity of thrombocytes, is apparently due to reduction of the surface glycoprotein GPIb, proved by flow cytometry and SDS-PAGE. The drop of glycoprotein GPIb to 20% of normal values suggests a heterozygote type of disorder with a medium grade of haemorrhagic manifestations. CONCLUSIONS: This is the first case of Bernard-Soulier syndrome in the Czech literature. Its analysis provides evidence of the relationship of the adhesive function to the surface glycoprotein GPIb and confirm the effectiveness of the elaborated diagnostic methods. PMID- 9221197 TI - [Ursodeoxycholic acid in the treatment of primary biliary cirrhosis]. AB - BACKGROUND: Pharmacotherapy of primary biliary cirrhosis (PBC) was not resolved unequivocally so far. During the last decade bile acids are used more widely. The objective of the submitted paper was to investigate under conditions of an open perspective study the influence of long-term administration of ursodeoxycholic acid on the clinical course and selected laboratory indicators. METHODS AND RESULTS: Ursodeoxycholic acid (Ursosan cps PRO.MED.CS) was administered to patients with PBC for a period of three years, 10-12 mg/kg/day. The investigation was completed by 13 women with a confirmed diagnosis of PBC which met clinical, laboratory and morphological criteria. During treatment marked improvement of itching was recorded, a significant drop of serum bilirubin, ALP and ALT. Changes of serum albumin levels were recorded only after three years treatment. The prothrombin time and galactose elimination capacity did not change significantly. Immunoglobulins M remained elevated and antimitochondrial antibodies were detected throughout treatment. The prognosis of the disease was evaluated by means of the Mayo score, the values of which declined significantly during the investigation. No serious side-effects were observed during treatment. CONCLUSIONS: Clinical evaluation proved beyond doubt a favourable effect of ursodeoxycholic acid on clinical and laboratory findings in PBC. Treatment should be started as soon as possible. Long-term continual administration is preferable. PMID- 9221198 TI - [Semiquantitative monitoring of molecular markers in non-Hodgkin's lymphoma]. AB - BACKGROUND: As quantitative changes of disease specific molecular markers may reflect disease activity, various methods quantifying targets of polymerase chain reaction were developed and their clinical relevance should be established. METHODS AND RESULTS: Here the exploitation of the DNA limiting dilution methodology in molecular monitoring of non-Hodgkin's lymphomas is presented. Long term stored diagnostic DNAs are checked for their integrity and examined in dose response assays for semiquantitative estimates of t(14,18) translocations and clonal immunoglobulin heavy-chain gene rearrangement. Assuming that the specific targets are diluted proportionally by dilution of total genomic DNA, sensitivities of polymerase chain reaction expressed as minimal amounts of total cells in reaction initiating positivity are compared. The term PCR-detectability as the ratio of sensitivities determined for preceding and actual samples is introduced. The value of PCR-detectability lower than one is considered as an indicator of a decrease of cells bearing the marker and vice versa. So far, the considerable increases in PCR-detectabilities were found close to relapses and unsubstantial changes at clinical remissions. CONCLUSIONS: It is presumable, that the semiquantitative limiting dilution approach may contribute to the monitoring of disease and treatment outcome. PMID- 9221199 TI - [Hypercalcemia as the first manifestation of bone marrow T-cell lymphoma]. AB - The authors describe the case of a 20-year-old patient where the first leading symptom was hypercalcaemia. A similar case was not published so far in the Czech literature. The disease took a fulminant course and proved fatal nine days after the first symptoms of the disease. The correct diagnosis was established only by necropsy. The adverse course of the disease could not be influenced by repeated haemodialysis nor by the administration of disodium pamidronate (Aredia) and calcitonin. The authors discuss differential diagnostic problems of hypercalcaemias and the pathogenesis of hypercalcaemia in malignant diseases of the haematopoietic system. PMID- 9221200 TI - [Diagnosis in environmental medicine: basic principles and problems]. AB - Diagnosis in environmental medicine only differs from the conventional medical diagnosis in a more detailed expositional evaluation on the basis of a respectively expanded anamnesis and a possible local visit, a surrounding examination as well as a so-called biological monitoring. Thus, the essential element of the "diagnosis" in environmental medicine consists in the resolving of a possible internal exposure (and occasionally resulting effects). From this point of view, physicians in environmental medicine could give an advisory contribution to the conventional medicine in selected cases. In contrast to frequently occurring assertions, there are practically no typical environmental diseases due to usual environmental toxicants. At present, a causality between environmental agents and health related disturbances can only be made plausible in less than 10% of out-patients of environmental medicine. These figures are in total contrast to the expansion of the out-patient and clinical environmental medicine. The expansion of the diagnostic offer gives not only to the public but also to the patients and the physicians the impression of a specific competence in diagnostic and therapy of environmental medicine which to this extent does not exist. The consequences are unnecessary and unsuccessful examinations. This is of no help to the patient. Most of the "environmental patients" suffer from civilization caused/psychosomatic and psychosocial disturbances like e.g. phobias, and somatoforme or depressive disturbances. In the genesis probable an increasing readiness for fear, unrealistic threatening convictions (arranged by media, homeopathists, physicians and other authorities), growing fear disturbances as a consequence to this as well as the cognitive connection of "normal" inner disturbances with the suspicious agens play a decisive role. For these patients the clinical environmental medicine lead astray. This is significantly more valid for the numerous "clinical ecologists" who apply scientifically doubtful methods. Considering the clinical approach to environmental medicine urgently needs a critical evaluation by independent research groups. PMID- 9221201 TI - [Possibilities for gathering environmental information for training environmental medicine specialists]. AB - In the context of continuing medical education, colleagues frequently face new professional topics. Advanced questions have to be clarified often to realize new skills. Beside the "conservative" information options like journals, literature searches in libraries and questioning colleagues, the "progressive" information procurement via CD ROM or the Internet is a comprehensive option to clarify certain questions quickly. PMID- 9221202 TI - [Pneumological aspects of environmental medicine. Medical relevance of environmental pollution of outdoor air (1)]. AB - The most important pollutants of industrial air pollution of the external air are sulfur dioxide (SO2), nitric oxides (NOX), oxygen radicals (ozone, O3) and sulfur dust. Brief detrimental effects on respiratory function of adults with healthy lungs are only observed during relatively high concentration of the pollutant, i.e. sulfur dioxide concentrations above 1500 micrograms/m3 or ozone 400 micrograms/m3. However, such a view of single pollutants is without medical relevance since all pollutants usually take effect as a pathological synergism in changing concentrations. Significant disturbances of the air ways may be found in sensitive persons (children, persons with air way problems, or cardiac and circulatory diseases) already after a slight increase in pollutant concentrations. It is assumed that the observed pollutant concentrations in the atmosphere close to the ground do not result in a permanent impact to health even under the circumstance of a combined effect. PMID- 9221203 TI - [Risk perception and risk communication in environmental medicine]. AB - Environmental medicine requires special communicative talents in order to inform about risks, to change risk-related behavior or to reassure people with excessive risk-related anxieties. We provide some guidance on how to accomplish these communication tasks. First, the results of psychological risk perception research are outlined, and the cognitive and affective factors, which determine laypeople's risk appraisal, are explained. Then, the basic problems of risk communication are described and the tasks and duties of medical risk communication are specified: How can risk information be indicated in an appropriate way? Which risk comparisons can be provided to enhance understanding? What are the advantages and drawbacks of risk comparisons? What criteria can be used to evaluate the quality of risk assessment studies? What are the Do's and Dont's of risk communication? And, how should anxieties about risks be approached? PMID- 9221204 TI - [Psychosomatic disorders in the area of environmental medicine. Environmental medicine--environmental psychology]. AB - In environmental medicine, we frequently see patients who have a very firm, sometimes fixated view of the nature of their disease, and they do not except the correction by the physician but only confirmation. Therefore, we face the task to undertake these patients a careful medical and psychological differential diagnosis. In a major number of cases, the symptoms are caused not by supported environmental effects but by an unknown diagnosis, and recognition and treatment would be impossible in case of an uncritical adoption of the patient's illness theory. Further, psychosomatic syndromes, which are well accessible by treatment procedures of psychosomatic medicine, can be diagnosed in many of those patients. This article demonstrates the different kinds of psychosomatic diseases in the area of environmental medicine and its appropriate therapeutic consequences. PMID- 9221205 TI - [Environmental concerns--fear of the environment or concern for the environment?]. AB - Increasing environmental pollution as reported by the media, makes people feel insecure and frightened which contribute to the onset environmental disease Environmental hazards and risks are perceived differently by lay people and by scientific experts. This is not a matter of irrationality of laymen. The layman's view of risks includes evaluatory and pragmatic (how to cope with the problem) dimensions. According to Kofler, toxicopy is regarded as a somatic reaction to a suspected threat by environmental pollution usually provoked by respective information by the media. Toxicopy is understood as a survival strategy under uncertain knowledge. Dealing with the fears of patients, the physician has to avoid enhancement of unfounded concern on one side and negating real problems on the other. High priority should be given to establish a trustful and co-operative therapeutic situation, in which the patients feels that his concern is taken seriously. PMID- 9221206 TI - [Exposure: environmental medicine considerations before measurements]. AB - Questions about the environmental impact on health result from the citizen's own perception, the information about exposure or from not yet explained diseases. They are usually related to staying indoor since one is there most frequently. To reach an immediate solution of such a conjecture, some people make use of measuring institutes. Unfortunately, medical advice is often requested afterwards when the inquiry is more difficult than expected. It makes more sense to consult a physician before the measurement. This article deals with environmental consulting for a suspected indoor pollution. It encourages considerations to make before a measurement to assign it a correct meaning in the context of medical diagnostics-even making it unnecessary. First thoughts are devoted to the medical view of environmental medicine since it mainly determines the access to the problem and the patient, and the patient himself. By structuring into the three categories PERSON, TIME, and LOCATION, the access to the complex question of indoor pollution may be simplified and "thinking of it" supported. In preparation for the visit of the patient's residence, the living conditions and the tenant's behaviour are examined. This structure is presented as a summary at the end of the article. PMID- 9221207 TI - [Status and perspectives of quality assurance in socialized medical services]. AB - Quality, quality assurance, and quality management are becoming more and more dominating topics in German public health. Quality is named as the most important goal of medical care in the public health related political discussions of physicians, hospitals, health insurances, and politicians but is affected by differences in political, economical, medical, and particular interests. To avoid a single-sided discussion about questions of quality (only with the goal of increasing efficiency and reducing costs), the society of physicians of Germany and the society of panel physicians published a common stock of 10 theses about medical quality assurance and quality improvement in the spring of 1996: (1) Medical quality assurance and quality improvement serve the patient. (2) Medical quality assurance and quality improvement do not primarily serve the improvement of economic efficiency. (3) Quality assurance programs have to be problem oriented and coordinated; quality in the ambulant and hospital setting cannot be different. (4) The appropriateness of quality assurance programs has to be evaluated firmly. (5) Transparency, communication, and cooperation are prerequisites of a successful quality assurance and quality improvement. (6) Comprehensive internal quality assurance is the foundation of continuous quality improvement. (7) External quality assurance should initiate the development of procedures for internal quality assurance. (8) Quality assurance has a chance for realization only if it is carried by the conviction and the effort of every participant to perform at relatively high quality, to subject his own doing to continuous checks and improvement, and to compare with others. (9) Quality has it's price. (10) Quality assurance and continuous quality improvement are the cornerstones of a quality policy in public health. The current situation and future developments of quality assurance in ambulant health care are discussed in this paper. PMID- 9221208 TI - [Quality assurance of inpatient care--exemplified by surgery]. AB - Internal quality assurance is natural for hospital physicians. External quality control is undertaken in cooperation with medical societies and sponsors-partly on voluntary basis. Despite considerable efforts in optimizing treatment procedures by classification of injuries and tumor stages, algorithms and tightly scheduled practical medical training, further control mechanisms are demanded; the economic drive is evident. In parallel to the introduction of quality control for services with a case-based flat rate, an interstructural result quality is desired. Hospital certification procedures are no longer utopian. The physician's duty for continuing medical education, which is set down in the professional code, has to be proven. The development of guidelines by scientific societies may help but is in need of a corrective. The medical chambers may play a particular role in this connection. Even if mechanisms of quality assurance differ, it is in the patient's interest that there must be no difference between ambulant and hospital care. PMID- 9221209 TI - [Quality in public health. Deficits, concepts and political quality key issues from the ministerial viewpoint]. AB - To preserve the quality of the German health care system as well as continuously optimize it towards the needs stated by ethics and law, an inter-professional and inter-institutional quality policy is required. It should be patient-centered, focus on process management and be based on EN ISO-Standards adapted to the specific needs of health care. The latter could provide internationally compatible models for quality management and quality improvement including economic efficiency. The 40 nation Council of Europe's 5th European Conference of Health Ministers in Warsaw as well as the 69th Conference of German Federal State Health Ministers (GMK) at Cottbus, who tackled the issue in November 1996, pointed out essential aspects. The GMK stated a lack of effective general concepts, quality control and patients' rights protection in Germany. Both conferences demanded equity, social justice and an active participation of patients in the setting of quality standards and the conception, functioning and control of health care. This includes rationalisations by using the limited funds in a most effective way. PMID- 9221210 TI - [Use of palm-top computers within the scope of medical data collection and quality assurance]. AB - Computerisation of the medical profession is advancing particularly as the reform of the German medical system is underway and efforts to maintain high quality standards are being made. More and more clinics and private physicians store the data, which they collect in their daily work, in digital form and exchange it via electronic networks. But lacking overall concepts and resistance from monopolistic structures prevent the introduction of innovative technologies, which would allow convenient data collection "on the fly", patterned after proven work routines. This could lead to a real relief from the daily paper work. New approaches using available hard- and software developed abroad demonstrate medical palmtop computing on the Apple Newton. PMID- 9221211 TI - [Quality assurance as the topic in medical periodicals. Results of a literature search]. AB - Against the background of the increasing importance of quality assurance procedures for the medical profession, it was questioned to which extend the topic "quality assurance" is content in medical journals. The frequency of articles with the key word "quality assurance" in German journals for the period from 1/1992 to 9/1996 was analyzed by using a literature search on Knowledge Finder's Health Star database. Journals for the clinical physician and the physician in the medical practice, that listed more than 200 articles, were taken into account. By using the key word "quality assurance", 276 articles were found (79 in 6 interdisciplinary journals, 197 in 22 medical journals). Interdisciplinary journals published between 8 and 25 (median: 13, rank 1: Zeitschrift fur arztliche Fortbildung), medical journals between 2 and 56 (median 13, rank 1: Chirurg). This analysis leads to the conclusion that a comprehensive continuing education about medical and interdisciplinary aspects of quality assurance in medicine only by reading common German medical journals may not be sufficient. PMID- 9221212 TI - [Graduate and continuing education from the viewpoint of Hamburg general practitioners]. AB - We wanted to investigate the interest and attitudes towards vocational training and continuous medical education (CME) among general practitioners in the city of Hamburg, Germany and carried out an semianonymous postal survey among all general practitioners (GP) in Hamburg excluding those specialised in psychotherapy using a pretested standardised questionnaire, 225 from 740 GP's responded (30%). More ground after vocational training was desired in the fields of "interpersonal competence", social medicine and specific topics from clinical medicine such as dermatology, psychiatry as well as musculosceletal diseases and complaints. Priorities for CME showed some significant differences with regard to gender and age of the GP's. Main topics for CME were categoried as "interpersonal competence", recognition and treatment of emergencies and pharmacotherapy. Priorities for CME are highly relevant to daily practice. Therefore, the chamber of physicians in cooperation with academic departments of general practice should offer more GP-specific and relevant topics. PMID- 9221214 TI - [Abstracts of presentations and posters from the 46th Pharmacologic Seminar in Olomouc (18-20 September 1996)]. PMID- 9221213 TI - [Cytochrome P450--its significance in the biotransformation of xenobiotics and interspecies comparisons]. AB - Enzyme system of cytochrome is the most important system of the oxidative biotransformation of xenobiotics. It was formerly assumed to be mainly a system of detoxication, however, it was recognized to be involved in many reactions leading to biological activation of compounds. This activation may give rise to unwanted products with rather detrimental effects. Isoforms of cytochrome are classified according to the similarity in primary structures into families and subfamilies. The great variability observed in enzymatic activities and in levels of particular isoforms is based on the known genetic polymorphism and on the enzyme induction or inhibition. However, this may be reflected in different pharmacologic responses to many drugs. Due to existence of interspecies differences in the presence of cytochrome P450 isoforms as well as due to differences in the substrate specificity it is difficult to extrapolate results obtained in experiments on animals to the human cases. A survey of isoforms, important for biotransformation of xenobiotics and of their properties is presented. PMID- 9221215 TI - [Malignant lymphoma with epithelioid cells]. AB - Authors studied a group of 6 cases of malignant lymphomas with epithelioid cells. Their additional common features were a variegated cell population, big admixture of T lymphocytes and rare elements reminding of Reed-Sternberg cells. The seventh case serving as a standard was Hodgkin's disease with a high content of epithelioid cells. According to phenotyping the group consisted of 3 peripheral T cell lymphomas of the type of angioimmunoblastic lymphadenopathy with dysproteinemia (AILD) and single cases of centroblastic ML, T-rich B-cell lymphoma and Hodgkin's disease. The latter diagnosis was settled after revision of a T-rich B-cell lymphoma. Some large cells were CD 20 and CD 30 positive. The classification was proved by autopsy. Authors tried to be more precise when classifying ML but they may be inapparent transitions among single types (e.g. between peripheral type of T cell lymphoma AILD type and AILD or between T-rich B cell lymphoma and Hodgkin's disease with lymphocytic predominance). The patients were followed for a relatively short period. Four of them died in several months after diagnostic excision, two showed a conspicuous generalization at autopsy. The presence of epithelioid cells in ML may not be connected with a more moderate behaviour and better prognosis. PMID- 9221216 TI - [Perineurioma]. AB - Perineurioma (storiform perineurial fibroma) is a rarely diagnosed benign tumour of perineurial cells. Presented tumour occurred subcutaneously in the right thigh of a 53-year-old man. It was well limited and consisted of whirl like and concentric laminar bundles of spindle and oval cells in variable local density. A storiform pattern was not present. Cells with richer cytoplasm, bland nuclei and solitary intranuclear pseudoinclusions formed focal meningioma like structures. Differential diagnosis had to distinguish benign neuronal tumours, myxoid variant of dermatofibrosarcoma protuberans and especially a "low grade" fibromyxoid sarcoma. A relevant immunohistochemical marker of perineurioma (for differential diagnosis) was the expression of epithelial membrane antigen (EMA) with negativity of S-100 protein. PMID- 9221217 TI - [Giant cell granulomatous aortitis and pulmonary arteritis]. PMID- 9221218 TI - [The ferricyanide reduction reaction as a staining method in liver biopsy]. AB - In paraffin sections from human liver tissue, ferric ferricyanide reduction reaction enables to visualize various reducing substances. Firstly it is cholestasis due to reduction capacity of bilirubin. Another reducing substance is lipofuscin pigment. Finally, the reaction represents the only way to demonstrate the needle-shaped cytoplasmic inclusions in routine practice considered to be specific for porphyria cutanea tarda. The method is simple and inexpensive and it may be recommended as a suitable supplementary staining procedure for liver biopsies. PMID- 9221219 TI - [Diagnostic algorithm for the evaluation of estrogen receptor function in breast carcinoma]. AB - Hormone receptor expression in breast cancer is not always a reliable reflection of their functional ability. This limitation affords a less valid prediction of tumor cell response to hormone treatment. For explanation of this phenomenon, the relationship between estrogen and progesterone receptor expression versus proliferative activity and the estrogen receptor related protein p29 expression was examined. Additional aim was to provide clues for interpretation of immunohistochemical findings from the point of view of the functional status of the estrogen receptor regulatory cascade. The results made possible establishment of a new diagnostic algorithm for evaluation of the estrogen receptor functional ability in breast cancer. PMID- 9221220 TI - [Superficial epithelial-stromal ovarian tumors in girls]. AB - Superficial epithelial-stromal ovarian tumours are unusual in adolescent girls (when compared with adult women) and extremely rare before menarche. A group of 40 girls with such tumours was chosen among 180 cases (22%) of ovarian tumours in childhood and adolescent age. All of them were between 9 and 18 years with one exception of a 5-year old girl. Mucinous tumours (22 cases) prevailed a bit over the serous ones (16 cases), two tumours were seromucinous. 35 tumours were one sided, 5 bilateral belonged to serous tumours. One sided tumours comprised 5 cystadenomas, 5 cystadenofibromas and 1 borderline serous tumour. Bilateral neoplasms were represented by 3 serous cystadenofibromas, 1 borderline serous tumour and 1 serous cystadenocarcinoma. Mucinous tumours comprised 16 benign cystadenomas, 4 borderline intestinal type tumours (one with identifiable eosinophilic indifferent cells) and 2 mucinous intestinal type cystadenocarcinomas. Both mixed tumours had the structure of seromucinous cystadenoma. PMID- 9221221 TI - 22nd Annual scientific meeting of the Australian Atherosclerosis Society. Perth, Western Australia, 9-12 October 1996. Proceedings and abstracts. PMID- 9221222 TI - Dental Enamel. Proceedings of a Symposium. London, 23-25 April 1996. PMID- 9221223 TI - [Antibiotic treatment in rheumatoid arthritis?]. PMID- 9221225 TI - [Hyperbaric oxygen therapy in sports injuries]. PMID- 9221224 TI - [Medical treatment in rheumatoid arthritis and mortality]. PMID- 9221226 TI - [Current aspects of alcohol-induced liver damage]. PMID- 9221227 TI - [Excimer laser treatment in corneal surgery]. PMID- 9221228 TI - [Rheumatoid arthritis shortens life]. PMID- 9221229 TI - [Adder bites are dangerous even in adults]. PMID- 9221230 TI - [Nephronophthisis--hereditary progressive renal disease]. PMID- 9221231 TI - [Respiratory symptoms in hantavirus infection]. PMID- 9221232 TI - [Spinal or epidural anesthesia?]. PMID- 9221233 TI - [Vulvodynia--sensitivity to touch of the vulva]. PMID- 9221234 TI - [Therapy-resistant pneumonia in a middle-aged man]. PMID- 9221235 TI - Proceedings of the 2nd International Conference on Animal Models for Aging Research. Kyoto, Japan, May 12-19, 1995. PMID- 9221236 TI - [Tick bite and sequelae: tick-borne encephalitis and Lyme borreliosis. And the tick continues to lurk...]. PMID- 9221237 TI - [Basic research is handicapped by lack of funding. Bioimplants of ceramics--a vision of the future?]. PMID- 9221238 TI - [Bioresonance therapy in treatment of allergies. Every person has his own vibration pattern. Interview by Beatrice Wagner]. PMID- 9221239 TI - [Beta blocker for therapy of heart failure. Decreased mortality and fewer clinic admissions]. PMID- 9221240 TI - [Pharmacotherapy of depressed patients. Increasing the therapeutic arsenal with a new generation of antidepressive drugs]. AB - Antidepressants form the core of the biological approach to antidepressive treatment. The present paper presents and discusses the basic principles of treatment with antipressants, for example selection criteria, dosage, maintenance therapy, etc. In particular new developments in the area of pharmacotherapeutic treatment are examined. In comparison with the classical tricyclic antidepressants, antidepressants of the third generation such as SSRI, selective and reversible MAO-A inhibitors, antidepressants with a dual mode of action acting selectively on specific neurotransmitter systems, seem to be better tolerated while offering comparable efficacy. PMID- 9221241 TI - [Suicide prevention--recognizing and treating acute suicidal behavior]. AB - Over the years, numerous models and working hypotheses have been developed in attempts to understand the underlying causes of suicide, and data on the frequency of certain risk factors are now available. Although a large number of institutions ranging from emergency telephone advice services to departments for crisis intervention are in place, the suicide rate remains distressingly high-in particular in certain high--risk groups such as patients suffering from chronic depression or an addiction. While physicians need to be aware that their influence too may be limited, they would do well always to keep in mind the particular risk factors and high-risk groups. PMID- 9221242 TI - [Diagnostic strategies in rheumatology. 4: Laboratory diagnosis--autoantibodies, immunogenetics and diagnosis of inflammation]. PMID- 9221243 TI - [Acute bronchitis: effectiveness of Sinupret. Comparative study with common expectorants in 3,187 patients]. PMID- 9221244 TI - [Chronopathology: from hypertension in general practice to manifest hypertension]. PMID- 9221246 TI - [Research on the Internet...or the art of finding a needle in a haystack]. PMID- 9221245 TI - [General practice 2000--what should be done today?]. PMID- 9221247 TI - [Children should enjoy coming to my practice]. PMID- 9221248 TI - [Recent changes and trends in angiographic techniques]. PMID- 9221249 TI - Recent Progress on the Molecular Aspect of Endocrine Tumors: Clinical Implications. Proceedings of the 39th Henri-Pierre Klotz Symposium on Clinical Endocrinology. Paris, France, May 30-31, 1996. PMID- 9221251 TI - Selected papers from the 5th Congress of the European Glaucoma Society. Paris, France, 20-22 June 1996. PMID- 9221250 TI - Consensus report. International Agency for Research on Cancer. Mechanisms of fibre carcinogenesis. PMID- 9221252 TI - [Cochleoscopy]. PMID- 9221253 TI - [NT-3 and BDNF for potential inner ear therapy]. PMID- 9221254 TI - [Hearing disorders in infants and young children. Possibilities for early diagnosis and therapy]. PMID- 9221255 TI - [Molecular biology in oncology of head-neck tumors]. PMID- 9221256 TI - [Nasal hyperreactivity]. PMID- 9221258 TI - [Guidelines/algorithms of the German Society of Otorhinolaryngology, Head and Neck Surgery]. PMID- 9221257 TI - [Nasal hyperreactivity. Allergic rhinitis and its differential diagnoses. Consensus report of pathophysiology, classification, diagnosis and therapy]. PMID- 9221259 TI - [Potassium ion secretion and generation of the endocochlear potential in the stria vascularis]. AB - Central to inner ear research are questions regarding the homeostasis of the high endolymphatic potassium concentration (approximately 150 mmol/l) and the high endocochlear potential (approximately +80 mV). Disturbances of the endocochlear potential can lead to the immediate loss of hearing which may be irreversible. The molecular mechanism leading to the generation of the endocochlear potential has not yet been discovered in spite of its clinical relevance. It is long known, however, that the stria vascularis is responsible for both the generation of the endocochlear potential as well as the secretion of potassium into endolymph. Recent investigations have clarified the mechanisms leading to the secretion of potassium and have led to the formulation of a now widely accepted model. This model explaining potassium secretion as well as the generation of the endocochlear potential is discussed in the present article. PMID- 9221260 TI - [Individual differences in susceptibility to motion sickness]. AB - Several different theories exist about the origin of kinetosis and the space adaptation syndrome, with individual sensitivities differing significantly. One explanation involves the hypothesis of a different otolith mass between the right and left statolith organ and especially a difference in the utricles. A difference in mass results in a different sensitivity to acceleration. For this reason we measured interindividual variances in saccular and utricular otolith mass. Since the anatomy of the vestibular organ in vertebrates is based as similar principles, we selected fish (salmon and trout) as our study model to facilitation preparations. The maximum difference in mass in the saccule was 17% and was generally smaller in the utricle, although in individual cases was much higher. We assume that a misbalanced sensitivity of the statolith organs occurs but is totally compensated for by the vestibular system as long as physiological motion patterns take place. Decompensation leads to kinetosis under non physiological motion patterns. When the vestibular system is better balanced and has an equally distributed otolith mass to both sides, the possibility for developing kinetosis or space adaptation syndrome is much less likely. PMID- 9221261 TI - [In vivo cochleoscopy through the round window]. AB - The overall aim of the present investigation was to develop a technique for endoscopic investigation of the cochlea. In the experiments reported here, the possible effect of the endoscope-called the "cochleoscope"-on the electrophysiology of the cochlea was investigated by recording the cochlear action potential (CAP) threshold tuning curve from (0.1-34 kHz). The dorsolateral bulla of anesthesized guinea pigs (with ketamine 60 mg/kg and Rompun 12 mg/kg) was opened, after which the cochleoscope was introduced under micromanipulator control through the round window membrane. Three cochleoscopes were used and had diameters of 0.29 mm, 0.7 mm and 0.89 mm, respectively, containing 2000, 3000 and 3000 fibers each. Experiments in 7 animals showed that the cochleoscope did not influence CAP thresholds. Although the present resolution of the endoscopes is limited, the basilar membrane can be clearly distinguished from the osseous spiral lamina. It is anticipated that improved resolution will allow the cochleoscope to be used for diagnostic purposes in cases of sensorineural hearing loss. PMID- 9221262 TI - [Ionomer cement as bone substitute in the middle ear of the rabbit]. AB - Ionomer-based cements are obtained by the reaction of an aluminum-fluoro-silicate glass with a polyalcenoic acid. During setting and hardening the cement bonds closely with adjacent hard tissue. The previous implantation of this material in the baboon tibia has held great promise as a possible use in bone replacement. In the present study the cement was tested concerning its biocompatibility and biostability in the middle ears of 64 rabbits. Viscid cement paste was inserted into the epitympanic space of each animal. A preformed cement strut was then placed to serve as a columella between the eardrum and stapes footplate. During a subsequent interval of 28 days up to 2 years middle ear specimens were evaluated under a surgical microscope, following which histologic sections were studied under light microscopic conditions. Findings demonstrated that after insertion of freshly mixed cement a firm adhesion to bone developed that proved to be biocompatible and biostable over time. After 28 days the preformed and fully hardened implants were overgrown by a delicate mucosa normally present in the middle ear. No evidence for any rejection of the implants could be found. The experience available to date indicates that ionomer cement is biocompatible and biostable, easy to handle and workable without splintering. With appropriate use it represents a useful implant material in surgery of the head and neck. PMID- 9221263 TI - [Optimizing hearing screening by transient evoked otoacoustic emissions in newborn infants]. AB - One of the major drawbacks using transient evoked otoacoustic emissions (TEOAEs) for hearing screening in newborn infants is the high fail rate in normal-hearing children. The purpose of the present study was to improve the overall performance of the test procedure and reduce of the fail rate. Improvement was obtained by 3 modifications: (1) change of the pass criterion in healthy newborns by requiring TEOAEs in at least one ear and nor both ears, (2) performing the test only after the second post-partum day, and (3) using a second-stage screening prior to hospital discharge in newborns who failed the initial test. In all, 3980 newborns from 2 well-baby clinics and 243 newborns from a neonatal intensive care unit (NICU) were screened using an system ILO-88 system program mode quick-check test was completed in 3820 infants. Considering these modifications, the pass rate of healthy newborns was improved from 79.5% to > 99%, and only 0.7% of the false negative (healthy) newborns required further audiological evaluation. Babies from the NICU failed in 3.8% of the screening tests. Overall, the number of newborns who failed the ILO-88 screening test was significantly reduced and the fail rate minimize when compared to previously published experiences with TEOAEs in new born hearing screening. PMID- 9221264 TI - [Endoscopic removal of osteoma of the paranasal sinuses]. AB - Osteomas are benign bone tumors, which are mostly localized in the nasal sinuses. The removal of nasal sinus osteomas, because of their hardness and scope, mostly requires extensive drilling work, which, especially in the case of big osteomas, also makes a large access path necessary. The introduction of endoscopic or microscopic technologies and experiences from the endoscopic opening of the osseous choanal atresias, where extensive drilling deep in the nose is carried out, make the endonasal access possible, even for the removal of nasal sinus osteomas. We treated a 59 year old patient, who, beside a pronounced polyposis of the nasal sinuses, also exhibited a large osteoma in the area of the right ethmoidal labyrinth. In the course of the intranasal sinus operation, the osteoma was completely removed via the same access route. The osteoma had to be gradually minced, intranasally, as it was too large to pass through the piriformes aperture. This approach shows an expansion of our endoscopic possibilities. Additionally, the etiology of the osteoma and its correlation with the polyposis is discussed. PMID- 9221265 TI - [Ophthalmoplegia. Ophthalmoplegia in sphenoid sinus metastasis of a laryngeal carcinoma]. PMID- 9221266 TI - [Management of cochlear implants in children. The Hannover concept]. PMID- 9221267 TI - The clinical pathophysiology of aldosterone. Introduction. PMID- 9221268 TI - Primary aldosteronism. AB - The basic clinical pathophysiology of primary aldosteronism (PAL) was described by Conn in terms of autonomous production of aldosterone, secondary suppression of renin and development of hypertension with hypokalaemic alkalosis. Conn recognised a normokalaemic form of the syndrome and suggested that it might masquerade as essential hypertension and be not uncommon. This was hotly disputed at the time, and normokalaemic PAL considered rare until recently, and, as a consequence, overlooked. The advent of a simple screening test, the aldosterone renin ratio, led to recognition that normokalaemic forms are not uncommon. In fact, PAL may be the commonest specifically treatable and potentially curable form of hypertension so far identified. In all patients with PAL confirmed by lack of suppressibility ("autonomy") of aldosterone production, Familial Hyperaldosteronism Type I (FH-I, glucocorticoid-remediable hyperaldosteronism, reviewed elsewhere in this issue) should first be excluded by dexamethasone suppression or genetic testing. Capable of causing fatal stroke in young people affected by this dominantly inherited disorder, it can be reversed by doses of glucocorticoids such as dexamethasone which partially suppress endogenous ACTH without producing "steroid" side-effects. The remaining varieties of PAL may eventually also be shown to have a genetic basis, but are currently treated either by excision of a solitary aldosterone-secreting tumour or by antagonism of aldosterone's action in the renal tubule. It is possible that both adrenal cortices are genetically predisposed to overproduction of aldosterone in all varieties of PAL, whether because of anomalous regulation of aldosterone secretion or because of a tendency towards hyperplasia and neoplasia. Aldosterone producing adenomas (APA's) can be divided into two main subtypes based on morphology and biochemical behaviour. The first subtype to be morphologically and biochemically characterised is composed predominantly of fasciculata-like cells and is unresponsive to angiotensin II (ALL-U-APA). The more recently characterised subtype is composed predominantly of glomerulosa-like cells, is responsive to angiotensin II (AII-R-APA) and could previously have been misdiagnosed as bilateral hyperplasia. The renin gene is often overexpressed in the second variety of adenoma, and in surrounding non-tumorous cortex, and the two subgroups show different allelic frequencies for RFLP's of the constitutive renin gene and the constitutive ANP gene locus. Unilateral, solitary, benign adrenal cortical adenomas producing aldosterone (APA's) represent a potentially surgically curable form of hypertension. Adrenal venous sampling (AVS) should always be performed because APA's are biochemically recognisable by adrenal venous steroid measurement before they are identifiable by computerised tomography or scintigraphy, and adrenal masses seen on CT may not be responsible for PAL. The secretory activity of adrenal masses must therefore be established by AVS before surgical removal. Discovery of an adrenal mass on CT requires formulation of a plan, whether or not it is found to be secreting hormones in excess. Independently of the treatment of the patient's hypertension, an apparently nonfunctioning adrenal mass ("incidentaloma") should be removed if 2.5 cm or more in diameter, because of the risk of cancer. Smaller masses require long-term follow-up. Primary aldosteronism not lateralising on AVS should be treated with low dose spironolactone, or with amiloride. For any such patients intolerant of medical treatment, laparoscopic removal of the adrenal showing higher production of aldosterone on AVS is an option worthy of consideration.The resultant reduction in mass of tissue autonomously secreting aldosterone should improve hypertension, as aldosterone productions falls below a critical level, and may even be curative in the short, medium or long term, depending on the rate of growth and activity of au PMID- 9221269 TI - Glucocorticoid-remediable aldosteronism. AB - GRA is an inherited disorder of aldosterone biosynthesis. To date, all cases have been the result of chimeric gene duplications in which the regulatory region of the 11-beta hydroxylase gene is fused to more distal coding sequences of the aldosterone synthase gene. This results in ectopic expression of aldosterone synthase in fasciculata cells. Genetic testing has been remarkably precise in identifying these individuals with 100% concordance of the presence of the chimeric gene with increases in 18-oxygenated cortisol products. Several implications follow from these findings. First, GRA may be more common in the hypertensive population than had been previously estimated, and second, genetic testing of subsets of the essential hypertensive population (e.g., those who have low plasma renin activity) may allow the identification of GRA patients who could then be treated specifically. We recommend that hypertensive patients with signs of aldosteronism and no radiologic evidence of an aldosteronoma, especially young hypertensive subjects with low renin activity, be genetically screened for GRA. To track the success of this approach and to identify responses to various therapeutic programs, a central international registry for GRA has been established. This registry not only provides access to screening for GRA, but also informational resources for patients and physicians. PMID- 9221272 TI - 17 Alpha-hydroxylase deficiency. PMID- 9221271 TI - Vascular remodeling and mineralocorticoids. AB - Circulating mineralocorticoid hormones are so named because of their important homeostatic properties that regulate salt and water balance via their action on epithelial cells. A broader range of functions in nonclassic target cellular sites has been proposed for these steroids and includes their contribution to wound healing following injury. A chronic, inappropriate (relative to intravascular volume and dietary sodium intake) elevation of these circulating hormones evokes a wound healing response in the absence of tissue injury--a wound healing response gone awry. The adverse remodeling of vascularized tissues seen in association with chronic mineralocorticoid excess is the focus of this review. PMID- 9221270 TI - Apparent mineralocorticoid excess syndromes. AB - Apparent mineralocorticoid excess (AME) is a syndrome attributable to congenital deficiency of the enzyme 11 beta-dehydrogenase (11 beta-OHSD) which converts active glucocorticoid cortisol to inactive cortisone. When 11 beta-OHSD activity is impaired, cortisol acts as a potent mineralocorticoid and causes hypertension and hypokalemia with a suppression of the renin-angiotensin-aldosterone system. The increased ratio of urinary cortisol/cortisone metabolites and a prolonged half-life of cortisol are useful for the diagnosis. Dexamethasone and/or potassium sparing diuretics have been used for medication of AME. Licorice ingestion induces a mineralocorticoid excess state, and it seems that this is the result of acquired inhibition of 11 beta-DH by glycyrrhetinic acid. The existence of a second 11 beta-OHSD isoform has been suggested strongly for a long time, and recently, a human 11 beta-OHSD 2 cDNA has been isolated. It appears that 11 beta OHSD 2 conveys specificity upon the renal MR, and a defect in its activity seems likely to account for the phenotype of AME. PMID- 9221273 TI - 11 Beta-hydroxylase deficiency. PMID- 9221275 TI - Expression cloning of type 2 angiotension II receptor reveals a unique class of seven-transmembrane receptors. PMID- 9221277 TI - The mineralocorticoid receptor discriminates aldosterone from glucocorticoids independently of the 11 beta-hydroxysteroid dehydrogenase. PMID- 9221276 TI - The biochemical phenotypes of two inborn errors in the biosynthesis of aldosterone. PMID- 9221278 TI - Vascular aldosterone. Biosynthesis and a link to angiotension II-induced hypertrophy of vascular smooth muscle cells. PMID- 9221274 TI - Glucocorticoid and mineralocorticoid resistance. PMID- 9221281 TI - Health care into the next century: markets, states, and communities. Part 1: Legacies and markets. PMID- 9221279 TI - Evidence that high dose cortisol-induced Na+ retention in man is not mediated by the mineralocorticoid receptor. PMID- 9221280 TI - Liddle's syndrome: heritable human hypertension caused by mutations in the Beta subunit of the epithelial sodium channel. PMID- 9221282 TI - Tuberculosis among homeless people at a temporary shelter in London. PMID- 9221284 TI - Prolonging death. PMID- 9221283 TI - Some misconceptions about understanding autoimmunity through experiments with knockouts. PMID- 9221286 TI - The CME scam. PMID- 9221285 TI - The need for preservation of quality in telemedicine: state licensure. PMID- 9221288 TI - Abstracts from the J. Douglas Miller Memorial Meeting. Edinburgh, Scotland, October 16-18, 1996. PMID- 9221287 TI - Shwachman Award acceptance. PMID- 9221289 TI - Auxiliary to the National Medical Association: its commitment to the National Medical Association. PMID- 9221290 TI - How to Build a Blood Vessel, Vascular Disease Symposium. Bethesda, Maryland, February 27-28, 1997. Abstracts. PMID- 9221291 TI - [Can superior cervical ganglionectomy be an experimental model of stellate ganglion block?]. PMID- 9221292 TI - Cancer Chemopreventive Agents: Drug Development Status II. PMID- 9221293 TI - Boston Working Group on Improving Health Care Outcomes Through Geriatric Rehabilitation. Proceedings from the conference. May 16-18, 1996. PMID- 9221294 TI - [Topoisomerase I inhibitors]. PMID- 9221295 TI - [Positron emission tomography]. PMID- 9221296 TI - [Early summer meningoencephalitis and Lyme borreliosis. Tick-borne diseases]. PMID- 9221297 TI - [Science entails responsibility. Opening address of the president of the 103rd Congress of the German Society of Internal Medicine, Wiesbaden, 6 April 1997]. PMID- 9221298 TI - [Changes in therapy of cardiac arrhythmias]. PMID- 9221299 TI - [Genesis, pathophysiology and clinical aspects of cardiac arrhythmias. Characterization of the risk patient]. AB - Extrasystoles and both supraventricular and ventricular tachycardias may occur as a complication of almost any underlying cardiac disease and many extracardiac causes; on the other hand, also a patient without any detectable structural heart disease may present with these arrhythmias. Refined mapping techniques of the intracardiac conduction process have let to important new informations about the pathophysiology of sustained tachycardias (focal impulse formation, macro reentry) with practical consequences, for example when ablation of these arrhythmias by radiofrequency catheter ablation is considered. Cardiac arrhythmias may lead to both typical and atypical symptoms. Finally, the patient at risk of sudden cardiac death is characterized. One needs to differentiate patients who have survived a life-threatening event of ventricular tachycardia or have been successfully resuscitated from cardiac arrest (both situations usually need life-long antiarrhythmic interventions for secondary prophylaxis) from patients who also are at high risk, however, are asymptomatic until now. For the latter population, symptoms due to extrasystoles or assessment of the severity of ventricular ectopic beats by the Lown classification are of minor importance; most emphasis in this regard, however, must be placed on the type and severity of underlying cardiac disease. PMID- 9221300 TI - [Drug therapy of tachycardic atrial arrhythmias]. AB - There are defined indications for the acute pharmacological management of AV nodal reentry tachycardias, AV reentry tachycardias and for the acute and chronic ventricular rate control in atrial fibrillation. Possible indications arise for the chronic pharmacologic therapy of AV reentry tachycardias, pharmacological cardioversion and prophylaxis of atrial fibrillation. In future there will be a trend towards nonpharmacological management of atrial arrhythmias. PMID- 9221301 TI - [Catheter ablation and implantable atrial defibrillators in supraventricular cardiac arrhythmias]. AB - Non-pharmacological tools for treatment of supraventricular tachycardias include radiofrequency catheter ablation, antiarrhythmic surgery, and electrical therapies. Radiofrequency catheter ablation is the first choice in the treatment of symptomatic patients with AV nodal reentrant tachycardias and atrioventricular reentrant tachycardias because of its high success rate and its low complication rate. Furthermore, transvenous radiofrequency catheter ablation may be considered as a curative approach in patients with atrial flutter and in patients with ectopic atrial tachycardias. Whereas the application of radiofrequency catheter ablation for the curative treatment of atrial fibrillation is still experimental, palliative therapy modalities such as AV nodal modification or AV nodal ablation may be performed in patients with drug refractory atrial fibrillation with rapid ventricular response. The recurrence rate of atrial fibrillation is in the range of 40 to 60% within a year despite antiarrhythmic drug treatment. Internal atrial fibrillation is a safe and effective method for acute termination of atrial fibrillation, especially after unsuccessful external cardioversion. The electrotherapy with the implantable atrial defibrillator should be considered as an alternative approach in patients with symptomatic, long lasting, and drug refractory episodes of atrial fibrillation. This innovative electrotherapeutic tool is currently under clinical evaluation. For the selection of the most appropriate therapy, the risk-benefit-ratio has to be taken into account in each individual patient. PMID- 9221303 TI - [Change in therapy of cardiac arrhythmias. Current studies--initial results]. AB - Treatment of Arrhythmias in Development-Recent Trials, First Results When dealing with preliminary or brand new results of large randomised trials, the requirement should be the assessment of the data by experts in the arrhythmia field. This holds especially true for studies addressing the complex problem of the prevention of sudden cardiac death. During recent years a number of controlled randomised trials have been published showing no or a harmful effect of antiarrhythmic drugs on the prognoses of patients at risk of sudden cardiac death. The results of these studies have clearly changed the scenario of preventive antiarrhythmic drug treatments. Some newer studies-published only in a preliminary form-seem to indicate that some Class III antiarrhythmics can have a positive effect. Studies are under way to compare the preventive effect of this antiarrhythmic drug therapy against sudden cardiac death with that of the implantable cardioverter/defibrillator. The improvement of prognosis remains the ultimate goal of antiarrhythmic therapy. PMID- 9221302 TI - [Drug therapy of ventricular arrhythmias]. AB - In patients with no or only mild structural heart disease, spontaneous ventricular ectopy which causes symptoms is being treated with beta receptor antagonists, sotalol, or in rare cases with class I substances. For primary prevention of sudden death, for instance in survivors of myocardial infarction, beta receptor antagonists are the only substances for which benefit has been demonstrated in large scale trials. In contrast, class I agents are contraindicated for this purpose. In secondary prevention of sudden death in patients with a history of sustained ventricular tachycardia or ventricular fibrillation, treatment with sotalol or amiodarone can be considered. However, nonpharmacological therapy by means of implantable defibrillators is increasingly applied in this patient population. PMID- 9221304 TI - [The concept of therapeutic equivalence]. AB - Usually, it is the purpose of a clinical trial to demonstrate the superiority of a (new) treatment in comparison to another treatment with regard to a well defined criterion of efficacy. However, other aspects rather than improved efficacy might be regarded as advantages of a new therapy, i.e. less or less severe adverse events, a more simple applicability, or a lower price. In this case, it may be sufficient to show a "comparable" efficacy (therapeutic equivalence). Unfortunately, equivalence studies can lead to severe problems of interpretation in case of insufficient methodological planning. In general, more detailed information must be available in advance compared to the common (superiority) trials. Very carefully designed trials are necessary to evaluate the therapeutic equivalence of treatments. PMID- 9221305 TI - [Definition of "chronic fatigue syndrome" (CFS)]. AB - The definition of "Chronic Fatigue Syndrome" (CFS) in 1988 was an attempt to establish a uniform basis for the previously heterogeneous approaches to research of this severe and inexplicable state of fatigue. At the same time, researchers wished to narrow down a pathogenetically founded disease entity a priori by specifying precise disease criteria. The empirical data gathered in accordance with the CFS definition, however, have failed to confirm the assumption that the disease entity is pathogenetically uniform. Furthermore, the originally selected criteria have proven to be impracticable ore theoretically questionable. In the period that followed, modifications that permitted a more comprehensive and yet more differentiated classification of fatigue states of unclear etiology were proposed. The new research approach avoids postulation of causal entities and puts CFS back in a category with other descriptive states of fatigue. PMID- 9221306 TI - [Clinical value of the CA 19-9 tumor marker with special reference to the Lewis phenotype]. AB - BACKGROUND: Because of structure and biosynthesis of CA 19-9, it was postulated that patients with the Lewis phenotype Le(a-b-) are not able to synthesize CA 19 9. But some patients with Le(a-b-) on red blood cells showed elevated levels of this tumor marker. PATIENTS AND METHOD: In 164 patients suffering from benign or malignant diseases both CA 19-9 and the Lewis phenotype were determined in sera. In addition in 51 patients red blood cells were tested for Lewis substances. RESULTS: The frequencies of the different Lewis phenotypes on red blood cells were compared with the results found in sera. The prevalence of the phenotype Le(a-b-) on erythrocytes was significantly higher than in sera. In 51 patients both determinations were performed. These results were compared additionally. The phenotype Le(a-b-) found on red blood cells agreed with the results found in sera only in 30% of the cases. A loss of Lewis substances on erythrocytes could be seen both in malignant and benign diseases. Only in patients with Lewis substances found in sera elevated levels of CA 19-9 could be seen. CONCLUSION: Considering only the Lewis phenotype in sera, it could be confirmed that patients with the genotype Le(a-b-)are not able to express elevated concentrations of CA 19-9. PMID- 9221307 TI - [Recurrent swelling of the knee joint. Synovial osteochondromatosis]. PMID- 9221308 TI - [Leiomyosarcoma of the esophagus. Clinical aspects, diagnosis and therapy based on an individual case]. AB - Leiomyosarcomas of the esophagus are rare tumors of mesenchymal origin. Apropos of a case we present clinicopathological features, diagnostic procedures and management of this seldom tumor of the esophagus. Primary gastrointestinal sarcomas cause less than 0.5% of all esophageal malign tumors, and present in ca. 5% as esophageal leiomyosarcomas. The most frequent incidence ranges between the fourth and fifth decade of life. The tumors originate from the muscular layers of the esophageal wall and are localized predominantly in the middle and distal third of the esophagus. Dysphagia is the most important and leading symptom although it presents late in the course of the illness. Endosonography is at the time the most accurate method to establish the tumor size. Differentiation between leiomyoma and leiomyosarcoma is only possible by histopathological examination and may be difficult in certain cases. Histopathological grading of the tumors as low- and high-grade sarcomas in dependence of the number of mitosis affects predominantly the prognosis of these patients. Differential diagnosis includes spindle cell carcinoma and carcinosarcoma of the esophagus. The most effective therapy consists in the complete operative removal of the tumor, in these cases five years survival rates of 30 to 40% are achieved, strongly influenced by tumor differentiation and size. PMID- 9221309 TI - [Prostate carcinoma associated spontaneous factor VIII:C inhibitor hemophilia. Successful therapy of severe hemorrhagic complication with porcine factor VIII in a 75-year-old patient]. AB - Inhibitors of factor VIII are a rare condition in non-hemophiliacs, but they are frequently responsible for life threatening hemorrhage. Acquired factor VIII:C inhibitors represent the spontaneous development of autoantibodies that partially or completely neutralize the plasma coagulant activity of the clotting factor. The autoantibodies can arise in diverse clinical settings, in older adults they are frequently associated with immunologic disorders or malignancies. We report of a 75-year-old man with acquired factor VIII:C inhibitor associated with adenocarcinoma of the prostate and a successful treatment of a severe bleeding complication with porcine factor VIII. A 75-year-old man was admitted because of a hematoma of his right cheek and an isolated prolonged aPTT. Acquired factor VIII:C inhibitor was identified as the cause and immuno-suppressive therapy was begun. In the clinical course severe hemorrhaging occurred and was successfully treated with porcine factor VIII (Hyate:C). The initially high inhibitor titer of 32 Bethesda Units (BU) disappeared. As the cause of acquired factor VIII:C inhibitor a newly diagnosed adenocarcinoma of the prostate is likely. After complete remission of acquired factor VIII:C inhibitor radiation therapy was begun. Six months after severe hemorrhaging the patient was clinically stable and PSA levels were normal. This case demonstrates the necessity of a precise diagnosis and therapy regimen of this coagulopathy based on clinical and laboratory data. In the absence of hemorrhage immuno-suppressive therapy with corticosteroids is indicated, in a patient with severe bleeding and high inhibitor titer (> or = 5 BU) porcine factor VIII should be administered. PMID- 9221310 TI - [Comparison between the diagnostic accuracy in diagnosis of thyroid nodules with fine needle biopsy an intraoperative histological evaluation of frozen tissue]. AB - BACKGROUND: The aim of this study was to compare the diagnostic accuracy of fine needle biopsy (FNB) and intraoperative frozen-section biopsy (FS) regarding the surgical management of thyroid nodules. METHODS: A total of 812 patients with solitary nodule or dominant nodule in a multinodular goiter were evaluated. The patients underwent preoperative FNB and intraoperative FS diagnosis. RESULTS: The definitive histological diagnosis (HD) was: i) 222 malignant lesions (118 papillary, 67 follicular, 16 anaplastic and 8 medullary cancers); ii) 590 benign lesions. FNB accuracy was 90.6%, sensitivity 96.8% and specificity 87.1%. FS accuracy was 97.4%, sensitivity 91.3% and specificity 100%. False negative (FN) were 10 for FNB and 21 for FS. False positive (FP) were 74 for FNB and 0 for FS. FS was less sensitive for the diagnosis of papillary cancer (more FN) and more specific for the diagnosis of follicular thyroid cancers (no FP). CONCLUSIONS: In conclusion, FS is useful in patients undergoing surgery for a thyroid nodule having a "suspicious" cytology. It adds no information in patients with an FNB diagnosis of malignancy and is of limited use in those in whom an FNB benign lesion is diagnosed. PMID- 9221311 TI - [Lipoprotein(a), parameters of lipid metabolism and hemostasis in obese patients]. AB - BACKGROUND: The prevalence of obesity is increasing in the population of industrialized countries and this condition is today considered a risk factor for cardiovascular diseases because, at least partly, it represents a prothrombotic state. METHODS: For this reason we have studied plasma lipid concentrations (C, TG, HDL-C), Lp(a) and some parameters of haemostasis (PAI-1, t-PA, D-dimer) in 41 non diabetic obese patients (38 females and 3 males) and in 36 healthy normal weight subjects. RESULTS: Lipid pattern has resulted in overlapping both in the two studied populations and in the two subgroups of pre- and postmenopausal women, while greater concentrations of Lp(a), (p < 0.001) and t-PA (p < 0.05) have been found in obese populations vs controls, but not different between the two subgroups; in particular, Lp(a) has resulted > 30 mg/dl in 34.14% of obese patients and in no case in the control group. Finally, Lp(a) concentrations have been found for the first time to be positively related to C and negatively to HDL C in the obese population. CONCLUSIONS: As Lp(a) is believed to be a pathogenetic linkage between atherogenesis and thrombosis and can be affected by weight loss, the efficacy of the reduction of high plasma levels of Lp(a) needs to be considered with longitudinal studies. PMID- 9221312 TI - [Acute hyperparathyroidism associated with follicular carcinoma in the thyroid: possible role of juvenile cervical irradiation. Description of a case]. AB - Acute onset of primary hyperparathyroidism is uncommon; neuropsychiatric signs are prominent clinical features in acute hypercalcemia and they can subside after normalization of serum calcium. Radiation therapy is a well-known risk factor for non medullary thyroid cancer, but it induces also parathyroid tumors. Data from the literature show that patients previously treated with neck radiation have an increased risk of primary hyperparathyroidism. Furthermore concomitant thyroid cancer is more frequent in radiation-induced hyperparathyroidism than in sporadic primary hyperparathyroidism. The case of a 63-year-old female patient who at the age of 14 had been irradiated to the neck for goiter and at the age of 50 had been repeatedly hospitalized for psychosis is presented. She was admitted to the hospital for suspected recurrence of psychosis, but clinical findings and urgent biochemical data showed on the contrary that she had a severe hypercalcemic crisis. Serum parathormone concentrations, neck echography and 99mTc-Sestamibi scintigraphy suggested hyperfunction of the right lower parathyroid gland; therefore the patient was operated on. Pathological examination disclosed a parathyroid adenoma but also two foci of follicular cancer in the right thyroid lobe with a metastasis to a lymph node were observed. Neuropsychiatric signs disappeared after normalization of calcemia and 6 months after operation the patient is free from psychiatric symptoms, despite she had stopped neurolectic drugs. It is underlined that patients who had received neck irradiation must be carefully observed because they are at increased risk of primary hyperparathyroidism and concurrent thyroid cancer. PMID- 9221313 TI - [Analysis of ocular saccadic movements with a fatigue test and neostigmine in Graves' ophthalmopathy]. AB - BACKGROUND: Aim of this study was to investigate the frequency of association between Myasthenia Gravis (MG) and Graves Ophthalmopathy (GO) using the responses to diagnostic tests for MG (fatigue and neostigmine tests) by measuring horizontal saccadic eye movements (SEM). METHODS: For this paper a random investigation was performed on eleven patients, affected by Graves' disease (GD) at the department of Molecular and Clinical Endocrinology and Oncology and department of Neurology of University of Federico II of Naples, for one year. Eleven patients (11 F), were subjected to endocrinological and ophthalmological examinations (TSH IRMA, TT3-RIA, TT4-RIA, TgAb, TPOAb, orbit ultrasonography or computed tomographic scans) and computerized analysis of saccadic eye movements (SEM); the fatigue and neostigmine tests were performed by means of SEM analysis in seven of this patients. RESULTS: Our results have reported that a positive response to the diagnostic tests for MG is present in 41.6% of hyperthyroid patients with GO, further supporting the hypothesis of a common autoimmune pathogenesis is present between these pathologies. CONCLUSIONS: SEM analysis may be a useful adjunct in the diagnosis of GO. PMID- 9221314 TI - Gillette children's special healthcare. For kids' sake. PMID- 9221315 TI - [A statistical contribution to fractures of the maxillofacial area]. AB - Twenty years experience (from 1975 to 1994), on 483 patients with injury of the maxillo-facial area observed at the Department of Maxillo-Facial Surgery of the University of Naples "Federico II" is reported. The analysis of the data, according to the literature, shows that the sex most involved is male (76%) with a prevalence of the second decade of life (38%); street injury is the most frequent cause of maxillo-facial fracture (48.5%), while in 1.6% the cause is not reported. Mandibular fractures (57%), particularly those of the left condyle, are more frequent. The type of treatment is discussed in relation to the site of fracture, in particular the therapy of the condylar ones. In fact, in the treatment of those fractures the authors use sequential functional plates, the first of whose presents a monolateral byte, homolateral to the injury, while the second a contralateral guide in closure. The use of such plates, even if it doesn't often obtain a perfect anatomical realignment of the fracture, allows an optimal functional rehabilitation due to the potential of remodelling of the condyle, avoiding complications related to surgical intervention. PMID- 9221316 TI - [Sternocleidomastoid-mandibular interaction and the rest position]. AB - BACKGROUND: The aim of this study was to evaluate the functional role of the interaction between the sternocleidomastoid muscles and the mandibular corners (ISM) in the context of the stomatognathic apparatus. METHODS: Registrations were effected about opening and closing movements of the mandible from the rest position with two different postures of the head. For the evaluations four volunteer subjects unaffected by functional anomalies of stomatognathic apparatus were selected. RESULTS: The resulting graphics allowed the elaboration of a theory about the way of setting up the mandibular rest position. The interactions between the sternocleidomastoid muscles and mandibular corners might be the supports where the strengths, which work on the mandible to avoid any stresses to masticatory muscles, are spread. The knowledge of determining factors for the rest position is considered basic for both diagnosis and rehabilitation purposes. CONCLUSIONS: Furthermore this study allows us to explain the already known relationships between the stomatognathic apparatus and cervical rachis with a new method. PMID- 9221317 TI - [Gingival hypertrophy due to cyclosporine. A clinico-statistical study in 82 patients]. AB - Eighty-two patients were observed at the Dental Department at Parma University. Seventy-six of them had renal transplant and 6 liver transplant. They were in immunosuppressive therapy with cyclosporina, 42 of them were also in calcium antagonist therapy. Gingival hypertrophy was observed in 52 subjects (63.4%) 25 (30%) patients underwent surgical operation, 6 of them (24%) showed a relapse about 3 months after the first operation. Clinical data were measured in accordance with 5 indicators: the level of oral hygiene, cyclosporinemia, contemporaneous use of calcium antagonist, the duration of therapy and the DMF index. By the results obtained, it's possible to suppose that the gravity of the disease is related to the contemporaneous use of calcioantagonist, but it wasn't highly significant (p < 0.05). The degree of oral hygiene was decidedly in relation to the most severe forms of gingival hyperplasia (grade 2-3), (p < 0.001). No relation was found for the duration of therapy, distribution of the DMF index and the level of drugs in the blood. PMID- 9221318 TI - [The treatment of the cardiac patient in dentistry and oromaxillofacial surgery. I. The practical management of patients with arrhythmias]. AB - Cardiac patients consist of a high incidence rate in odontostomatology, both clinical and surgical. Moreover this serious complication disease conditions odontostomatological and, particularly, surgical works. In this article the authors present the results of several years of research carried out to obtain a correct clinical and therapeutic approach for clinical and surgical dentistry. After an introduction on the clinical features of heart diseases the most important clinical cases of heart dysrhythmia are discussed: like, i.e. hypokinetic arrhythmia, hyperkinetic arrhythmia and the management of patients with pacemakers. The principal diacritic features of dysrhythmic diseases are illustrated. Anxiety is a sort of disease not directly related with dysrhythmia. Moreover a lot of clinical studies find in heart arrhythmia the principal problem caused by anxiety on heart physiology. Consequently the authors describe anxiety in the same part of pathologies commonly known as heart dysrhythmia. In the last phase the authors illustrate the most opportune therapeutic steps corresponding to the principal pathologies described above. These matters were dealt with from an odontostomatological point of view. The results obtained suggest the necessity of keeping to the management that was described. Actually a low percentage of accidents occurred only when the above-mentioned clinical processes were completely performed. PMID- 9221319 TI - [The treatment of the cardiac patient in dentistry and oromaxillofacial surgery. II. The practical management of patients with hemodynamic pathologies]. AB - When odontostomatological or surgical treatment is performed we suggest, in a first phase, to distinguish cardiac patients from the others. In a second phase a careful nosological diagnosis will be performed. Consequently, patients' medical history plays a fundamental role in both diagnostic phases. In this article the authors present the results of several years of research carried out to obtain a correct clinical and therapeutic approach for clinical and surgical Odontostomatology. After an introduction on the clinical features of heart hemodynamic pathologies the most important clinical cases are discussed: like, for example, acardiohemia, valvulopathies and heart decompensation. The principal diacritic features of hemodynamic diseases are illustrated. Essential hypertension (borderline and resident) is a sort of disease not directly related to hemodynamics pathology. Moreover a lot of clinical studies find in heart hemodynamic pathologies the principal problem caused by hypertension on heart physiology. Consequently the authors describe essential hypertension in the same part of pathologies commonly known as heart hemodynamic pathologies. In the last phase the authors illustrate the most opportune therapeutic steps corresponding to the principal pathologies above-described. These matters were dealt with from an odontostomatological point of view. The results obtained suggest the necessity of keeping to the management that was described. Actually a low percentage of accidents occurred only when the above-mentioned clinical processes were completely performed. PMID- 9221320 TI - [Dental changes induced by radiotherapy: a study of subjects with retinoblastoma. I]. AB - The authors examine the dental and skeletal alterations consequent to the use of radiotherapy as the elective method of treating retinoblastoma, an endobulbar neuroectodermal tumour typical of early infancy. The authors revise the basic concepts of radiobiology and the modifications induced by radiation in developing tissues with reference to both alterations at a microscopic (histological) level and those at a macroscopic level, namely of a morphological and structural nature. In the first part of the study the authors examine the damage to dental structures relating to both developing tissues and damage to mature tissue. The second part of the paper analyses radiation damage on cartilage and bone tissues in the process of formation and the authors present their documentation in the form of iconographic and radiographic material together with an esthetic face analysis performed using Ricketts' [correction of Rickettz's] method in three cases referred to their attention. PMID- 9221321 TI - [Augmentation of the maxillary sinus by Le Fort osteotomy in endosseous implantology. A clinical case report]. AB - The authors present a case of sinus floor elevation using Le Fort I osteotomy. They underline that, owing to its peculiarities, this type of intervention is indicated, in their opinion, for sinus floor elevation when both the vertical dimension and possible skeletal discrepancies require modification. They briefly describe the indications, peculiarities and contraindications. PMID- 9221322 TI - [Primary Ewing's sarcoma of the mandible. A clinico-pathological and immunohistochemical case study]. AB - A rare case of Ewing's sarcoma, originating in the mandible, is reported. The symptomatologic and radiological aspects is often aspecific. For this reason the diagnostic-therapeutic routine is presented and the difficulty of clinical diagnosis is accentuated. In these cases it may be appropriate to make use of immunohistochemical analysis such as the research of markers like the NSE (Neuro Specific Enolase). This will be useful to reach an accurate preoperative diagnosis and in order to adopt a correct therapeutic protocol. PMID- 9221323 TI - Red blood cell transfusions contaminated with Yersinia enterocolitica--United States, 1991-1996, and initiation of a national study to detect bacteria associated transfusion reactions. AB - Although bacteremia and sepsis are infrequently reported complications of red blood cell (RBC) transfusion, receipt of transfused blood contaminated with bacterial pathogens may result in sepsis, disseminated intravascular coagulation, and death. Such pathogens have included Yersinia enterocolitica and Pseudomonas fluorescens. From November 1985 through February 1991, a total of 11 cases of sepsis associated with receipt of transfused Y. enterocolitica-contaminated RBCs were reported in the United States. This report describes an additional 10 cases of Y. enterocolitica sepsis reported to CDC during March 1991-November 1996 in patients who received transfusions with contaminated RBCs and describes the development of a study to detect bacteria-associated reactions to transfusion of RBCs and other blood components. PMID- 9221324 TI - Introduction to Table V. Premature deaths, monthly mortality, and monthly physician contacts--United States. AB - Beginning with this issue, a new table will appear monthly in the MMWR:"Table V. Potential Years of Life Lost, Deaths, and Death Rates, by Cause of Death, and Estimated Number of Physician Contacts, by Principal Diagnosis" (see page 557). By displaying a variety of measures that gauge the importance and relative magnitude of certain public health issues, this table will call attention to those issues where strategies for prevention are needed. Publication of this table reflects CDCs increased responsibility for promoting action to reduce unnecessary morbidity and premature mortality and continues the MMWR's tradition of disseminating public health information to its readership. PMID- 9221325 TI - Status of public health--Democratic People's Republic of Korea, April 1997. AB - During 1995 and 1996, severe flooding in the Democratic People's Republic of Korea (i.e., North Korea (DPRK) (1990 population: 22 million)) (Figure 1) caused 186 deaths, dislocated approximately 550,000 persons from their homes, and caused damage to an estimated 1.2 million metric tons of crops (12% of total production). In combination with systemic economic problems in DPRK, these natural disasters have been associated with reports of a severe, ongoing food shortage and increased risks to public health. To assist in targeting humanitarian aid, in April 1997, the U.S. Agency for International Development's Office of U.S. Foreign Disaster Assistance requested CDC to conduct an onsite assessment of the public health status and needs of the DPRK. This report summarizes findings of the assessment, which indicate a recent substantial decline in the health and nutritional status in DPRK. PMID- 9221326 TI - Update: syringe-exchange programs--United States, 1996. AB - As of December 1996, approximately one third (36%) of the 573,000 cases of acquired immunodeficiency syndrome (AIDS) among adults reported to CDC were directly or indirectly associated with injecting-drug use. Syringe-exchange programs (SEPs) are one of the strategies for preventing infection with human immunodeficiency virus (HIV) among injecting-drug users (IDUs). The goal of SEPs is to reduce the transmission of HIV and other bloodborne infections associated with drug injection by providing sterile syringes in exchange for used, potentially contaminated syringes. This report summarizes a survey of U.S. SEPs regarding their activities during 1995 and 1996 and compares the findings with those during 1994 and early 1995. The findings indicate continued expansion in the number and activities of SEPs in the United States. PMID- 9221327 TI - Transmission of hepatitis C virus infection associated with home infusion therapy for hemophilia. AB - Transmission of hepatitis C virus (HCV) and other bloodborne viruses between household members who are not sex partners presumably results from inapparent percutaneous or permucosal exposures, such as sharing articles that may be contaminated with microscopic quantities of blood. The risk for nonsexual household transmission is extremely low, and no cases of such transmission have been documented; direct percutaneous exposures (e.g., injecting drugs) have been identified as the major risk factor for infection. This report summarizes the investigation of a newly acquired case of HCV infection in a child with hemophilia, after a preliminary investigation identified several household members with HCV infection. The findings suggest the child acquired infection through percutaneous exposure to the mother's HCV-infected blood during infusion of clotting-factor concentrate. PMID- 9221328 TI - Hepatitis A vaccination programs in communities with high rates of hepatitis A. AB - In June 1995, the Public Health Service Advisory Committee on Immunization Practices (ACIP) issued recommendations about the use of hepatitis A vaccine for the prevention and control of hepatitis A. In communities with high rates of hepatitis A and periodic outbreaks, the ACIP recommends routine vaccination of young children and catch-up vaccination of previously unvaccinated older children. This report describes hepatitis A vaccination programs initiated to control ongoing outbreaks and prevent future outbreaks in two communities with high rates of hepatitis A. Preliminary epidemiologic data indicate that the program in one area may have decreased the magnitude and duration of a predicted outbreak. The incidence of hepatitis A in other areas will require long-term monitoring to determine the effect of the vaccination program . PMID- 9221330 TI - Progressive Supranuclear Palsy (PSP) Europe International Workshop. Oxted, Surrey, England, October 25, 1996. Abstracts. PMID- 9221329 TI - Recommendations for follow-up of health-care workers after occupational exposure to hepatitis C virus. AB - Hepatitis C virus (HCV) infection is a major cause of chronic liver disease in the United States and worldwide. At least 85% of persons with HCV infection become chronically infected, and chronic liver disease with persistently elevated liver enzymes develops in approximately 70% of all HCV-infected persons. Persons with chronic hepatitis C are at risk for cirrhosis and primary hepatocellular carcinoma. Most HCV transmission is associated with direct percutaneous exposure to blood. Health-care workers (HCWs) are at occupational risk for acquiring this viral infection. However, no vaccine is available to prevent hepatitis C, and immune globulin is not recommended for postexposure prophylaxis. PMID- 9221331 TI - Leonard I. Malis. An appreciation. PMID- 9221332 TI - John E. Moseley, M.D. 1909-1996. PMID- 9221333 TI - Cellular telephones and traffic accidents. PMID- 9221334 TI - Cellular telephones and traffic accidents. PMID- 9221335 TI - Cellular telephones and traffic accidents. PMID- 9221337 TI - Digoxin in patients with heart failure. PMID- 9221338 TI - Digoxin in patients with heart failure. PMID- 9221339 TI - Digoxin in patients with heart failure. PMID- 9221340 TI - Digoxin in patients with heart failure. PMID- 9221341 TI - Racial variation in the use of coronary-revascularization procedures. PMID- 9221342 TI - Racial variation in the use of coronary-revascularization procedures. PMID- 9221343 TI - Treatment of ostial renal-artery stenoses with vascular endoprostheses. PMID- 9221344 TI - Phytanoyl-coenzyme A hydroxylase deficiency -- the enzyme defect in Refsum's disease. PMID- 9221345 TI - Pfizer Night at Boston Billiards. PMID- 9221346 TI - Bone mass and the risk of breast cancer. PMID- 9221347 TI - Bone mass and the risk of breast cancer. PMID- 9221348 TI - Bone mass and the risk of breast cancer. PMID- 9221349 TI - Bone mass and the risk of breast cancer. PMID- 9221350 TI - Hormone-replacement therapy and coagulation. PMID- 9221351 TI - Hormone-replacement therapy and coagulation. PMID- 9221352 TI - Screening patients with insulin-dependent diabetes mellitus for adrenal insufficiency. PMID- 9221353 TI - Detection of familial primary pulmonary hypertension by genetic testing. PMID- 9221354 TI - More on continuous-infusion acyclovir for severe varicella. PMID- 9221355 TI - Predispositions to meningococcemia. PMID- 9221356 TI - Psychiatric outpatient services in the United States and Canada. PMID- 9221357 TI - Psychiatric outpatient services in the United States and Canada. PMID- 9221358 TI - Psychiatric outpatient services in the United States and Canada. PMID- 9221359 TI - [Multidisciplinary treatment of superior sulcus (Pancoast) tumors]. AB - Ten patients with a Pancoast tumour, seven with pulmonary carcinoma, three with a soft tissue tumour, were treated surgically with or without preoperative chemotherapy or external radiotherapy, and with postoperative external radiotherapy mostly in combination with brachytherapy using a flexible intraoperative template. The results were highly variable, e.g. one patient died after three months, another was still alive without tumour after 36 months. Optimal treatment requires cooperation of experienced surgeons, radiotherapists and medical oncologists. PMID- 9221360 TI - [Medical and administrative neglect of high blood glucose levels; comments on a decision by a medical disciplinary tribunal]. AB - A 41-year-old man died in 1995 during ketoacidotic coma. He suffered from chronic manic depression, used lithium carbonate, and consulted the psychiatrist and the general practitioner (GP) frequently. Diabetes had not been diagnosed. Late in 1994 the situation worsened, the patient complaining of general illness, fatigue, nausea, vomiting, diarrhoea, thirst and excessive drinking of soft drinks. The GP referred the patient to a neurologist who found no neurological disorder but who asked for determination of blood glucose and lithium levels, and of thyroid function. The day afterwards the neurologist went on holiday. The blood glucose level proved to be elevated (16.9 mmol/1) but nobody took any action and the GP was not informed. Six days after returning from his holiday, the neurologist who had an administration backlog, found the laboratory findings only after he had been informed that the patient had just died. The court gave the neurologist a warning. Lessons are that somatic problems should be treated as such, even in a psychiatric patient, and that a good administrative signalling system is a prerequisite for quality in medical practice. PMID- 9221361 TI - [Assessment of spontaneous motor activity in young infants: an effective method for the detection of brain function disorders]. AB - A method for the assessment of the brain function of young infants was recently introduced. It consists of evaluation of the quality of spontaneously generated generalized movements (general movements, GMs). GMs appear at an early stage of pregnancy and persist until approximately the 4th month after term. Normal GMs are characterized by the triad of complexity, variation and fluency. Mildly abnormal GMs. indicating mild dysfunction of the nervous system, are not fluent but jerky or stiff. Markedly abnormal GMs, indicating major nervous system dysfunction, are characterized mostly by absence of complexity and variation of the movements: the movements are monotonous and stereotyped. The quality of the GMs can be evaluated by means of so-called global Gestalt perception. The technique can be learned in a few days. The quality of the GMs has a clear predictive significance for the child's development. Children with normal GMs will be free of handicaps in later life, whereas three-quarters of the children showing clearly abnormal GMs throughout the postnatal GM period do develop handicaps. Assessment of the quality of the GMs is a relatively cheap, non invasive method of evaluating the current and future brain function of young infants. PMID- 9221362 TI - [Transdermal opioid administration: the pain plaster]. AB - A new method of administration of an opioid was recently registered: fentanyl transdermal (brand name: Durogesic), intended particularly for the indication range 'pain in cancer'. Fentanyl is lipid-soluble so that deposition in the skin takes place and the biological half-life is approximately 20 hours after removal of the plaster. It is safe to start on a basis of an equianalgesic conversion of 100:1 in relation to oral morphine, although this may entail some risk of fentanyl under dosage. The dose adjustment time is 12-24 hours before a constant fentanyl level is reached; therefore, after attaching the first sticking plaster, the original morphine dose should be continued for another 12 hours. In addition, the patient may, if necessary, be given supplementary morphine preferably as a short-acting drug. There seems to be no clear indication for transdermal fentanyl either in neuropathic pain or in chronic benign pain. PMID- 9221363 TI - [Laxative policy for terminal patients ineffective]. AB - OBJECTIVE: To determine the prevalence of constipation and the use of laxatives in terminal patients. DESIGN: Retrospective. SETTING: Hospice Rozenheuvel, Rozendaal, the Netherlands. METHOD: Of patients who were admitted for terminal care between 1 January 1995 and 15 July 1996 the anamnestic data were recorded about presence of constipation and use of medicines, notably opioids and laxatives. RESULTS: The study population (n = 121) consisted of 65 male and 56 female patients. 95 patients (79%) suffered from cancer. The median survival time was 18 days (spread from 1-180 days). 58 (48%) of the patients were constipated at the time of admission and 62 (51%) used opioids. The opioid users were more often constipated (68% versus 27%; p < 0.001). 35 out of 62 (56%) opioid users were given laxatives. The laxative users, however, were more often constipated (65% versus 36%; p < 0.01). Neither in users nor in non-users of opioids did laxatives have any effect on presence of constipation. Of the laxative users 37 (73%) were given lactulose in a fixed dosage. CONCLUSION: Present laxative policy in terminal patients is ineffective. Monotherapy with fixed lactulose doses is not effective as a laxative in either users or non-users of opioids. PMID- 9221364 TI - [Acetylcysteine in children with lung disorders prescribed by one-third of family physicians: no support in the literature]. AB - OBJECTIVE: To determine whether the use of acetylcysteine in children is supported by literature data and to determine how often and for what indications acetylcysteine is prescribed for children in general practice. DESIGN: Systematic literature review and general practitioners audit. SETTING: Academic Hospital Groningen, the Netherlands. METHODS: A Medline search was performed and the references of the articles found were checked. All 720 general practitioners working in the three northern provinces of the Netherlands were mailed a questionnaire regarding their prescription of acetylcysteine for children. RESULTS: Of the studies on acetylcysteine in children with pulmonary disorders (excluding cystic fibrosis) (n = 15) the majority (n = 12) were uncontrolled clinical observations. Three clinical trials were found, all of which showed considerable methodological shortcomings. The observed benefit of acetylcysteine therapy in these studies was of no clinical relevance. The questionnaire was filled out completely and returned by 70.3% of general practitioners. Almost one third of the general practitioners (32.6%) prescribed acetylcysteine now and again for children with various pulmonary disorders (such as 'mucus that is difficult to bring up' (73.9%), asthmatic bronchitis (50.3). bronchitis (40.0%), excessive mucus production (40.0%) and dry cough (34.5%)). CONCLUSION: Acetylcysteine is being prescribed frequently for children with various pulmonary disorders by general practitioners whilst the use of this drug is not being supported by literature data. PMID- 9221365 TI - [AIDS epidemic in The Netherlands: current developments in transmission route, age and nationality]. AB - OBJECTIVE: To document the recent developments in the course of the AIDS epidemic in the Netherlands, 1982-1995. DESIGN: Descriptive. SETTING: National Institute of Public Health and Environment, Bilthoven, Municipal Health Service, Amsterdam, and Inspectorate of Public Health, Rijswijk, the Netherlands. METHOD: Based on the new AIDS patients reported to the Inspectorate of Public Health, the incidence figures were calculated by risk group (homo/bisexual men, intravenous drug users and heterosexual men and women), by birth cohort defined by 5 successive years of birth, and by nationality, in order to characterise sub epidemics. RESULTS: Among homo/bisexual men AIDS incidence has been stabilizing in recent years. Among intravenous drug users and heterosexuals incidence continues to rise but at a low level compared with homo/bisexual men. Among homo/bisexual men and intravenous drug users mean age at AIDS diagnosis is rising in conjunction with reduced incidence among young persons born in 1965-1969 when compared with the incidence among persons born in 1960-1964 when at the same age. By contrast, among heterosexuals a decline in mean age at diagnosis is observed and this decline coincides with undiminished rise of incidence among persons born in 1965-1969. Among heterosexual patients an increasing and disproportionate number have the nationality of a sub-Saharan African country. CONCLUSION: Future AIDS incidence among homo/bisexual men and intravenous drug users will probably be lower than it currently is. Regarding heterosexuals the undiminished growth of the number of young AIDS patients and the increasing proportion of patients from abroad make such an assessment more difficult. PMID- 9221366 TI - [High-flow priapism: a rare, easily treatable disorder with excellent prognosis]. AB - Four patients, men aged 33, 37, 37 and 12 years, were examined because of priapism following trauma. In all four high-flow priapism was diagnosed, based on intracavernous blood gas analysis and selective angiography. One of the men aged 37 was subsequently found to be suffering from low-flow priapism caused by chronic myeloid leukaemia (the blood gas analysis had been performed after decompression of the cavernous body). Two other patients were treated by selective internal pudendal artery embolisation. In the fourth, who developed a vascular spasm at angiography, embolisation was not performed: he recovered spontaneously. In contrast to high-flow priapism, low-flow priapism is an urological emergency for diagnosis and treatment to prevent permanent impotence. PMID- 9221367 TI - [Effective removal of certain skin pigment spots (lentigenes) using the Q switched ruby laser]. PMID- 9221368 TI - [Several patients with memory disorders]. PMID- 9221369 TI - [Favorable results of plasmapheresis in severe myasthenia gravis]. PMID- 9221370 TI - [The geographic distribution of tick bites and erythema migrans in The Netherlands]. PMID- 9221371 TI - [Arthroscopic reconstruction using the Leeds-Keio ligament following anterior cruciate ligament lesions in 200 patients]. PMID- 9221372 TI - In need of direction. PMID- 9221373 TI - The greatest gift is to share your knowledge. PMID- 9221374 TI - [Pathology and pathogenesis of viral hepatitis]. AB - Several factors viral genotype, mutations, virus-host interaction, expression of viral proteins and host immune-reaction are very important in the pathogenesis of hepatotrop viral infections. Activation of Fas/Fas ligand system occurs during hepatitis which is responsible for the apoptosis of virus infected hepatocytes. The histological features in chronic hepatitis can be classified based on the activity of near inflammatory alterations ("grade"). Stage of chronic hepatitis describes the degree of fibrosis and architectural alterations. PMID- 9221375 TI - [Management of chronic hepatitis B]. AB - The features of chronic hepatitis B virus (HBV) related liver diseases and the aim of their therapy have briefly discussed, then treatment modalities are listed. In Hungary, between 1994 and 1996, a total of 68 patients with chronic hepatitis B have been treated with interferon (IFN). IFN resulted in complete clinical-biochemical remission in 50% of the patients, and in 32% the HBV replication was also eliminated. There are various nucleoside analogues, among them mostly famciclovir and lamivudine have been intensively studied as potentially effective treatment for HBV infection, and controlled clinical trials are in progress with these drugs. Nucleoside analogues in combination with IFN possibly improve treatment results in this disease. Various immunomodulatory agents--such as levamisole, thymosine, interleukin-2, and other cytokines--as well as the prednisolon-withdrawal induced rebound phenomenon have also been tested in HBV infection, but with no generally established benefit. A recombinant HBsAg vaccine is under investigation for therapeutic use. For end-stage HBV liver cirrhosis, liver transplantation is the only treatment, but the problem of reinfection is not still solved for more reasons. PMID- 9221376 TI - [Chronic hepatitis C: virologic and immunologic aspects and questions of therapy]. AB - Characteristics of hepatitis C virus (HCV), importance of its genotypes and mutant variants "quasi-species", as well as the mechanisms of disease chronicity and tissue injuries caused by HCV have been discussed. Both a presumed direct cytopathy of the virus and the host's immune response may play a role in the pathogenesis. HCV infects not only hepatocytes but lymphoid cells, thereby modulates immune functions. A molecular minicri--that is a cross-reaction between viral and host antigens--also contributes to autoimmune phenomena developed during chronic HCV infection, in addition a genetic predisposition for autoimmunity can be involved. Until now the only accepted therapy for HCV infection is interferon, that at a standard low dose regimen, results long-term benefit only in one-fourth of treated patients. In Hungary, between 1994-1996 601 chronic hepatitis C patients have been treated: biochemical remission (ALT normalization) was found in 38% of the patients and elimination of the HCV was achieved in 25%. Several questions are to be answered concerning the optimalization of the treatment, e.g. dosage, duration of treatment, re-treatment of relapse, problem of non-responders, the role of predictors of response in the individualized therapy, usefulness of combination treatment. The original end points and goals of therapy are sustained HCV-RNA negativity, normalization of serum GPT/ALT, and inactivity in liver histology, yet the real end-points are incidence and prevention of cirrhosis and hepatocellular carcinoma, that is the better survival. Concerning these questions, newer therapeutic experiences are discussed, including results from Hungarian researchers. PMID- 9221377 TI - [Optimal antiviral therapy of chronic hepatitis caused by hepatitis C virus]. AB - Chronic HCV disease is a major health problem world-wide. The majority of HCV positive patients will develop progressive liver disease with a high risk for cirrhosis and hepatocellular carcinoma after 20 and 30 years respectively. Sustained response after a 6, month course of interferon therapy is about 30%. New insights in viral biology and virus-host interactions led to new guidelines for diagnosis and therapy. Seventy percent of anti-HCV positive persons with persistently normal ALAT levels are HCV-RNA positive, and 60-70% of these have chronic hepatitis histologically. Biochemical parameters alone are inadequate for diagnosis of the disease and evaluation of therapy. HCV-RNA positive patients with normal serum ALAT and chronic hepatitis histologically should be considered for antiviral therapy within the setting of clinical trials. Response rates to interferon have been doubled with 1 year treatment. Additional improvement may be achieved by higher dosage, especially in patients with hepatitis caused by genotype 1b viruses. Including viral and host parameters in therapeutic strategy may lead to selective adaptation of dosage in order to optimize interferon therapy and to further improve its cost-effectiveness. PMID- 9221378 TI - [Possibilities and perspectives of the prevention of viral hepatitis]. AB - Six district hepatotrop viruses causing viral hepatitis have been identified. Hepatitis A and E viruses are enterically transferred with feco-oral transmission, the others (hepatitis B, C, D and G viruses) produce the infection parenterally with blood, blood products and body fluids. All the hepatitis viruses are able to cause acute hepatitis. Chronic carrier state and chronic hepatitis can develop in case of infections with hepatitis B, C, D, and G viruses. In the prophylaxis the hygienic rules should be applied in all forms of infections. Passive immunisation and active vaccination have been safety developed til now only in hepatitis A and B infections. To prevent the hepatitis C, D, E and G infections the modalities of prophylaxis are in experimental stages. PMID- 9221379 TI - [Mesothelioma. Pathology/pathogenesis/mesothelioma register]. PMID- 9221380 TI - [Absence of association of specific and nonspecific bronchial hyperreactivity in occupational asthma caused by platinum salts]. AB - There is evidence that bronchial responsiveness to allergen is quantitatively correlated with bronchial responsiveness to nonspecific stimuli in subjects with allergic asthma. This association has been questioned in occupational asthma due to low molecular weight substances. It was the aim of this study to assess the quantitative association of bronchial responsiveness to methacholine (MCH) and platinum salts (Pt), in the form of hexachloroplatinic acid, in workers with occupational asthma due to Pt salts. Fifty-seven subjects with exposure to Pt, work-related asthma, and a positive bronchial challenge with Pt underwent skin prick test with Pt and bronchial challenge with MCH. Using the provocation concentration causing a > or = 50% fall in specific airway conductance (PC50sGaw(Pt)) as a dependent variable, anamnestic data (period from first symptoms to removal, period between removal from exposure and diagnosis, and smoking), season of the investigation, skin prick tests with environmental allergens, total immunoglobulin E (IgE), skin reactivity to Pt (Pt concentration causing a 2 mm wheal), and PC50sGaw(MCH) were included as independent variables for regression analysis. Fifty-two subjects (91%) showed a PC50sGaw(MCH) < 8 mg.mL-1 (geometric mean for all subjects 1.6 mg.mL-1). Responsiveness to Pt varied widely between subjects (geometric mean of PC50sGaw(Pt) 9 x 10(-5) mol.L 1, range 2 x 10(-7) to 10(-2) mol.L-1). There was no univariate correlation between bronchial responsiveness to MCH and Pt, but there was a correlation between skin reactivity to Pt and PC50sGaw(Pt) (r = 0.6). This association could not be improved by considering PC50sGaw(MCH), the period from first symptoms to removal, or the period between removal from exposure and diagnosis. The parameters that showed the highest (negative) associations with PC50sGaw(Pt) were skin reactivity to Pt and the period between removal from exposure and diagnosis (r = 0.65). We conclude that there is a moderate association between bronchial responsiveness to platinum salts and skin reactivity to platinum salts. However, there is no association between methacholine responsiveness and bronchial responsiveness to allergen in occupational asthma due to platinum salts. PMID- 9221381 TI - [Evaluation of the validity of spiroergometric parameters in the differentiation of circulatory and ventilatory physical limitation]. AB - BACKGROUND: One of the indications postulated for cardiopulmonary exercise testing (CPX) is the distinction between ventilatory and cardiocirculatory reasons of an exercise limitation. Though data such as anaerobic threshold, breathing- and heart-rate-reserve, difference of arterial and alveolar oxygen pressure, ratio of ventilation to O2 and CO2 output and aerobic capacity are commonly considered in CPX interpretation, exact choice of parameters and cut-off points is not definitely clear. The aim of our study therefore was to scrutinize the validity of these parameters and the recommended cut-off-points. A second goal was to evaluate the reliability of a standardized CPX interpretation as postulated by Wassermann and Palange. PATIENTS AND METHODS: We examined 19 patients with congestive or ischemic cardiomyopathy, 24 subjects with chronic obstructive or interstitial lung disease (n = 11 resp. n = 13). The control group consisted of 15 healthy people. During incremental cycle exercise we measured the ventilatory frequency, the peak O2 uptake and the oxygen pulse as well as the above mentioned parameters. Data were interpreted according to Wassermann and Palange in order to confirm the diagnosis of a cardiocirculatory or ventilatory conditioned exercise limitation. Additionally we determined the cut-off-points with the highest diagnostic value for our patient groups. RESULTS: According to Palange only 7% of the diagnosis could be confirmed, 55% were correctly ascertained by data interpretation according to Wassermann. Breathing reserve (45% of maximum voluntary ventilation) proved to be the only parameter with satisfying selectivity (sensitivity = 75%; specificity = 95%). Diagnostic value of all other recorded data was worse by far. CONCLUSIONS: Our study shows that standardized interpretation of CPX data does not lead to the correct diagnosis in a satisfying percentage. In any case results have to be confirmed by conventional tests such as ECG or spirometry, so that the additional diagnostic value of CPX in patients with an exercise limitation of unknown origin has to be seen very critically. Except breathing reserve from our point of view there is still no valid parameter or cut-off-point to distinct cardiocirculatory from ventilatory exercise limitation that can be recommended. PMID- 9221382 TI - [Comparison of nasal and bronchial production of nitric oxide in healthy probands and patients with asthma]. AB - Nitric oxide (NO) appears to play an important role in the pathophysiology of airway diseases as suggested from measurements of NO in exhaled air, animal and in vitro experiments. As NO is produced in variable amounts within the bronchial system and the nose, we studied the relationship between nasal and bronchial production of NO in patients with asthma and determined to which extent these productions were increased compared to healthy subjects. The nasal and bronchial production rates of NO as a function of breathholding time were assessed in 10 healthy subjects, 7 patients with asthma without inhaled corticosteroids, and 5 patients with asthma and a therapy of inhaled corticosteroids. After a breathhold of 10 s bronchial NO concentrations were elevated in the patients with asthma without steroids by the factor 3.5 (p < 0.005) and nasal concentrations by the factor 1.2 (n.s.) compared to healthy subjects. NO concentrations increased with time. Correspondingly, bronchial production rates were increased by factor 2.7 (p < 0.01) and nasal production rates by factor 1.1 (n.s.) in asthmatic compared to healthy subjects. The asthmatic patients with steroids showed lower production rates than those without steroids. We conclude from these data that in patients with asthma as compared to normal subjects bronchial production of NO is markedly increased, whereas the corresponding relative increase in nasal production is lower. PMID- 9221383 TI - [Pleurodesis in malignant pleural effusion: bleomycin vs. mitoxantrone]. AB - In a controlled clinical trial we investigated 102 patients with malignant pleural effusion due to breast cancer, lung cancer, ovarian cancer and other tumors to compare the therapeutic effect and adverse events of pleurodesis with bleomycin (BMC) or mitoxantrone (MIT) via chest tube. Finally 96 patients had been treated according to the protocol. Age, gender, Broca index, performance score or distribution of primary tumors were not statistically different between the BMC (n = 49) or MIT group (n = 47). We found no differences between intention to-treat and according-to-protocol groups as well. 30 days after BMC pleurodesis we found remissions of the effusion in 91% of patients (complete remission [CR] 51%, partial remission [PR] 40%), after 90 days in 83% (40% CR, 43% PR). 30 days after MIT instillation we found remission in 73% of patients (35% CR, 38% PR), after 90 days in 61% (29% CR, 32% PR) (30 and 90 days: p < 0.05). Adverse events were not different between BMC and MIT group. BMC is a safe and effective sclerosing agent for pleurodesis via chest tube. PMID- 9221384 TI - [Pentoxifylline inhibits experimental bleomycin-induced fibrosing alveolitis]. AB - Therapy of idiopathic pulmonary fibrosis (IPF) is directed at 1) inhibition of alveolitis and tissue damage, and 2) inhibition of matrix deposition. We and others have identified pentoxifylline (POF) as a promising drug in achieving these aims. For further clarification, we established a model of bleomycin induced fibrosing alveolitis. Fisher 344 rats (n = 7 per group) were given bleomycin intratracheally once (0.7 U/100 g bw) and treated with POF (1.5 or 3 mg/kg bw per day i.p.), prednisolone (15 mg/kg bw i.m. per day), or sodium chloride solution (NaCL). The extent of inflammatory reactions was determined after 8 days by differentiation of cells of broncho-alveolar lavage (BAL) and by quantification of proliferating cells in lung interstitium subsequent to staining of the Ki-67 antigen. POF inhibited neutrophil alveolitis in BAL and reduced the amount of proliferating cells in the lungs significantly while prednisolone and NaCL did not. Both POF and prednisolone exerted a positive influence on postoperative weight loss as well as on lung weight increase subsequent to bleomycin instillation. The postoperative body weight loss and the lung weight increase after bleomycin instillation are most likely due to an inflammatory reaction subsequent to operation and bleomycin deposition. Tumor necrosis factor alpha (TNF-alpha) has been shown to be a key cytokine in bleomycin-induced fibrosing alveolitis as well as in IPF; it also exerts catabolizing effects. Since both POF and prednisolone are known to effectively inhibit proinflammatory cytokines and, among those, TNF-alpha, nonspecific antiinflammatory effects probably explain the benefits. Additionally, however, this study proved POF to be more effective in inhibition of BAL neutrophils and number of proliferating cells in lung interstitium. Further, it has been shown that POF, but not prednisolone, inhibits activation of neutrophil granulocytes and formation of reactive oxygen species. Thus we believe that the mechanism of action of xanthines might contribute to therapy of IPF. For further clarification, a prospective clinical study of POF in IPF therapy has been initiated. PMID- 9221386 TI - Etiology of Breast and Gynecological Cancers. Proceedings of the 9th International Conference on Carcinogenesis and Risk Assessment. Austin, Texas, November 29 - December 2, 1995. PMID- 9221385 TI - [Cell biology of bronchial asthma: interesting new methods. Intracellular determination of cytokines in T-cells and basic proteins in eosinophilic granulocytes using flow cytometry]. AB - Clinical studies on pathophysiological mechanisms of asthma bronchiale require methods of investigation that are simple, reproducible and rapid in respect of analysing relevant inflammatory cells and mechanisms. The article presents two new flow cytometric methods enabling on single-cell level the determination of expression of intracellular cytokines in T-cells and of basic proteins in eosinophilic granulocytes. Both techniques of examination can be performed either with whole blood or with cells from bronchoalveolar lavage without preceding cell separation techniques. First results reveal interesting new aspects with regard to the expression of IFN-gamma, IL-2, IL-4 and IL-5 in T-cells, and of ECP and EPO in eosinophils in bronchial asthma. PMID- 9221387 TI - The life and work of Benjamin Bjorn Rubinstein. PMID- 9221388 TI - Psychoanalysis and the philosophy of science. Collected papers of Benjamin B. Rubinstein, M.D. PMID- 9221389 TI - On metaphor and related phenomena (1972) PMID- 9221390 TI - Benjamin B. Rubinstein's contributions to the structure of psychoanalytic theory. PMID- 9221391 TI - Psychoanalytic theory and the mind-body problem (1965) PMID- 9221392 TI - Psychoanalytic hypotheses and the problem of their confirmation (1978) PMID- 9221393 TI - Why metapsychology? (1978) PMID- 9221394 TI - On the psychoanalytic theory of unconscious motivation and the problem of its confirmation (1980) PMID- 9221395 TI - Freud's early theories of hysteria (1983) PMID- 9221396 TI - The experience of tragedy, expectations, and the moral order (1983) PMID- 9221397 TI - On the psychoanalytic concept of sexuality (1952) PMID- 9221398 TI - Explanation and mere description: a metascientific examination of certain aspects of the psychoanalytic theory of motivation (1967) PMID- 9221399 TI - [Proceedings of the 1st Hispanic Congress of the Health-Related Professions. Puerto Rico, 8-10 October 1996]. PMID- 9221400 TI - [Academic responsibility for public policy on health]. PMID- 9221401 TI - [Supply and demand in the health-related professions]. PMID- 9221402 TI - [Ethical theories and bioethical principles]. PMID- 9221403 TI - [Active patient participation in health-related decision making]. PMID- 9221404 TI - [Social marketing: a strategy for promoting the health-related professions]. PMID- 9221405 TI - [Practical hints on thoracic radiology]. PMID- 9221406 TI - [Quantitative analysis of radiologic progression in rheumatoid arthritis: controversies and perspectives]. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease with symmetrical polyarthritis as the major feature. Persistent inflammation leads to largely irreversible joint damage which can be seen radiographically. Radiographs depict the progression of joint damage and alterations, which are one of the major parameters of RA evolution. Delayed radiologic progression is a good indicator of the success/failure of long-term drug treatment, but the quantitative analysis of changes over time and the reliability of scoring systems remain difficult steps. This paper focuses of the main current scoring systems, with an emphasis on the following four: Larsen method and its modification by Kayle, Sharp method and its modification by van der Heijde. The scoring method--be it grading, counting, or weighted counting--did not appear to influence reliability or repeatability, while the radiologist's training and the film reading technique were critical to identify disease progression accurately. Our data consistently suggest the paired reading method as the most suitable for radiologic progression assessment in RA. The radiologic studies of 62 patients with early RA after 7 years' follow-up showed joint damage in 82% of patients. The average annual progression rate of the total radiologic score obtained with Sharp method, summing erosions and joint space abnormalities, was faster in the earlier years of the disease (8.8 units/year) than later on (4.9 units/year) (p < 0.01). From the trialist's point of view, these results imply that disease duration is a critical feature for RA treatment outcome. PMID- 9221407 TI - [Clinico-radiologic considerations on "stress fractures" of the leg]. AB - Stress fractures are ubiquitary and most often caused by the subject's activities. In the past they occurred mostly in recruits, but today they are frequent in sportsmen. Stress fractures most frequently occur in the lower limbs, especially in the distal leg. We reviewed 32 injuries observed January, 1993, to June, 1995, and found that 25% of them had been misdiagnosed as stress fractures: in the cases where the diagnosis was correct, fatigue fractures (32%) were less frequent than insufficiency fractures (68%) and occurred in young subjects (mean age: 24 years), usually sportsmen (2/3 of cases). Insufficiency fractures may occur in people aged 8 to 81 years (mean: 61 years) and in subjects with metabolic disorders (45.5%). Considering the injury biomechanics and the patient history and symptoms, these lesions appear a rather uncommon event, whose radiologic diagnosis must be confirmed by clinical findings, since radiology mostly (81.6% of cases) showed only the repair process, rather than the fracture itself. The radiologic patterns were classified into three groups: the fracture margin was not shown in 70% of cases (group I), where however intraperiosteal reaction and/or soft tissue effusion were found; bone fracture was shown in 3 cases (group II) and fracture sequels in 4 (group III), where bone thickening (3 cases) or abnormal consolidation (1 case) was found. There are several synonyms of "stress fracture" and confusion is increased because stress lines and other not necessarily abnormal signs such as Park or Harris lines, reinforcement or calcification lines, are often grouped together with stress fractures. Only accurate clinical examination and laboratory findings permit to distinguish fatigue from insufficiency stress fractures and the latter are also very difficult to differentiate from pathologic fractures. The differentiation of fatigue from insufficiency fractures, originally made by English speaking authors, may be confusing because the definition "pathologic fractures" should be reserved only to focal injuries while in the past it included also insufficiency fractures. Thus, only (bone) fatigue injuries in patients exercising intensely and constantly should be considered stress fractures. Conventional radiography is an indispensable tool and MRI is used in selected cases where the former method is negative and in the patients needing early mobility to go back to work. If radiographic findings are questionable for metastases, nuclear medicine is the method of choice and CT and/or MRI may be indicated as second-line diagnostic imaging tools. PMID- 9221408 TI - [Ultrasonic diagnosis of De Quervain's stenosing tenosynovitis]. AB - This study was aimed at stressing the role of US of the first dorsal compartment of the wrist as the initial diagnostic approach in the study of De Quervain's syndrome, thanks to the high diagnostic yield of this technique. We report on 14 cases of De Quervain's stenosing tenosynovitis whose onset of the symptoms ranged 3-24 months before the US exam. The patients were subdivided into three groups, according to disease stage and US patterns; 5 patients were in group 1, six in group 2 and 3 in group 3. The most frequent US findings were: thickening of the tendons, which appeared as separate structures or as a single structure (pseudofusion); diffuse or, in most cases, focal thickening of the sheaths; changes in tendon echogenicity; impaired tendon sliding, which was reduced or absent due to fibrous adhesions and compression by the sheaths. Diffuse tendon and sheath thickening was prevalent in group 1, where the onset of symptoms ranged 3-10 months earlier. Focal sheath thickening and tendon pseudofusion were prevalent in group 2, where the onset of symptoms ranged 8-15 months earlier. Tendon degeneration and fibrous adhesions between tendons and sheaths, as well as the US changes observed in group 2, were found in group 3 where the onset of symptoms ranged 18-24 months earlier. These US changes were not influenced by medical therapy, with the exception of local injections of corticosteroids. To conclude, we propose a classification of De Quervain's syndrome into three stages, according to the US signs characterizing the different stages of disease evolution and the time of onset of the symptoms. PMID- 9221409 TI - [Dynamic magnetic resonance of the knee. Considerations on techniques and anatomy with a magnetic resonance system with open magnet]. AB - MRI is currently considered the best technique to study the joints--knee joints in particular. Most studies are performed with the knee straightened out or slightly flexed, which position is mandatory with contemporary MR units. Recently, however, the MR equipment for limb studies and the larger tunnels of total body magnets have permitted partially dynamic knee studies, up to 30-40 degrees flexion; this joint excursion cannot be exceeded. In 1996, Muhle and Niitsu demonstrated that dynamic MRI can be useful to study cruciate ligament injuries, some types of meniscal injuries, all femoropatellar conditions and, mostly, cruciate ligament reconstruction. We used an open magnet MR unit for thorough functional dynamic knee studies, acquiring the images at 0-120 degrees flexion. The open magnet MR unit permits free joint movements. We examined 25 subjects: 10 healthy volunteers and 15 patients with meniscal, capsuloligamentous, femoropatellar or synovial conditions. This technique schedules axial, coronal or sagittal images, according to the diagnostic suspicion, suitably directed with the patient in lateral decubitus and with the knee flexed to maximum joint excursion. Dynamic MR findings are easily transferred to an X-ray film, so that the radiologist can send them to the orthopedist with no need of any videotape or TV take. The current cine MR programs display the dynamic images of the moving joint on an MR monitor, but this option is not yet widely available. The quality of dynamic MR studies is exactly the same as that of conventional images, with a particular emphasis on the small diagnostic details of the injuries. We report our preliminary experience with this technique, indicating some possible clinical applications of dynamic MRI which permits thorough studies of knee flexion-extension. PMID- 9221410 TI - [Assessment with magnetic resonance of cartilage injuries of the knee with tridimensional techniques with fat suppression]. AB - MR studies of chondral injuries of the knee are performed to obtain a high C/N ratio between the articular cartilage and such other structures as subchondral bone, intraarticular fat pad and synovial fluid. This goal has been achieved with the 3D SPGR fat-suppressed technique. This kind of sequence yields high spatial contrast resolution because of its contiguous thin (1.5 mm) slices and high contrast resolution between the hyaline cartilage, which is strongly hyperintense, and the surrounding structures (fat, synovial fluid, subchondral bone) which appear hypointense. From September, 1994, to February, 1995, we submitted to MRI 34 patients, adding the 3D fat-suppressed technique after the routine sequences. These volumetric sections were obtained to image the tibiofemoral joint cartilage in 15 patients and to study the patellofemoral compartment in 19 patients. Chondral injuries were graded according to Beguin and Locker classification and MR results were compared with arthroscopic findings. 3D SPGR fat-suppressed sequences permitted accurate chondral thickness evaluation and the depiction of cartilage injuries, especially in advanced injuries (stages 3 and 4). However, in earlier stages, this MR technique tends to overestimate globular hypointense areas in normal thickness cartilage. This MR pattern needs a close correlation with clinical findings and should be interpreted according to the presence/absence of other associated intraarticular injuries. PMID- 9221411 TI - [Mammography changes associated with hormone replacement therapy in post menopausal patients]. AB - Hormone replacement therapy (HRT) in post-menopausal or ovariectomized women reduces mortality due to cardiovascular diseases, lowers morbidity due to osteoporosis and improves vasovagal symptoms. Long-term therapy, however, increase the risk of side-effects. HRT may decrease mammographic sensitivity, markedly increasing glandular density. Enlargement of pre-existing cysts and fibroadenomas has also been reported after HRT. The correlation between HRT and breast cancer is highly controversial. We examined 650 women: 550 of them (84.6%) received HRT (157 estrogens and 393 estrogens-progestins) and 100 (15.4%) refused treatment and were thus considered as a control group. All patients underwent mammography and DEXA before HRT and, during treatment, were followed-up yearly with mammography, often combined with US, and DEXA. Fisher's test was used for data analysis (confidence interval: 95%). The statistical analysis showed a significant difference between the HRT group and the control group only for the lumbar spine. Mammographic changes (Tab. II) were shown in 150 of 550 HRT patients. Increased breast density was the most frequent finding. Benign lesions arising de novo or increasing in size and/or number were observed in 41 of 150 patients (27.3%) in the HRT group, where 3 breast carcinomas were detected, versus 1 breast cancer only in the control group. HRT had a marked positive effect on bone mineral content (BMC) at 2 years' follow-up, but it remains debated if it reduces breast cancer risk. In conclusion, our results indicate that a yearly mammography is mandatory in long-term HRT subjects and US may be also needed in particularly dense breasts. PMID- 9221412 TI - [Magnetic resonance angiography in stenosing-occlusive diseases of the carotid arteries: 3D with time of flight versus 3D with phase contrast]. AB - We assessed the comparative sensitivity, specificity and diagnostic accuracy of 3D time of flight (TOF) versus 3D phase contrast (PC) MRA in the study of stenoses/occlusions of the extracranial carotid artery. Fifty-four patients were submitted to MRA because of their symptoms or of signs of cerebrovascular insufficiency. 3D TOF and 3D PC axial slices were acquired with a high field magnet; digital angiography was the gold standard. The parameters of 3D TOF acquisitions were: TR/TE/FA 50/8/20, 1 NEX, 512 x 256 matrix, 16 x 12 FOV, 1 axial slab of 60 slices, 1 mm slice thickness, superior presaturation band, ramp pulse, TA 10-18 minutes. 3D PC parameters were: TR/TE/ FA 25/9/20, 1 NEX, 256 x 128 matrix, 18 x 13 FOV, 1 axial slab of 60 slices, 1 mm slice thickness, VENC 35 cm/s, TA 10-15 minutes. 3D TOF MRA was in agreement with digital angiography in 95.6% of cases (22/23), overestimating a grade 3 stenosis as grade 4 (one case, 4.3%) and underestimating no stenoses. 3D PC MRA was in agreement with digital angiography in 78.2% of cases (18/23), overestimating three stenoses (13%); one grade 1 as grade 2 and two grade 4 as grade 5; two stenoses were underestimated (8.6%); one grade 4 as grade 3 and one grade 4 as grade 2. Sensitivity, specificity and diagnostic accuracy were 91.4%, 98.9% and 95.7% for 3D TOF, versus 83.2%, 97.2% and 92.3% for 3D PC, respectively. The two MRA techniques had different semiology. In spite of outstanding background noise suppression, 3D PC poorly depicted the turbulent flow at the carotid bifurcation in carotid stenoses. Therefore, 3D TOF appears a better technique to study stenoses/occlusions of the extracranial carotid arteries. PMID- 9221413 TI - [Usefulness of the modified Dixon technique associated with gadolinium administration in the characterization of adrenal masses]. AB - We investigated the usefulness of modified Dixon technique at 0.5 T after Gadolinium (Gd) administration in the characterization of adrenal masses. One hundred and one patients (45 men, 56 women; mean age: 60.5 +/- 13.5 years) with 125 adrenal masses found at previous US or CT of the abdomen were submitted to MRI. The study protocol included preliminary T1-weighted, PD and T2-weighted SE images. A bolus of 0.1 mmol/kg Gd-DTPA was administered and then T1-weighted SE scans were acquired with modified Dixon technique (TR = 450-600 ms, TE = 30 ms, 192 x 192 matrix, no craniocaudal presaturation). Acquisition time is twice as long with this technique as with conventional SE sequences, but three sets of T1 weighted images are acquired: conventional SE, fat-suppressed and water suppressed images. The diagnosis was made at surgery (18 lesions) and fine-needle biopsy (34 lesions), or with stable US follow-up findings for at least 1 year (73 masses). Adrenocortical adenomas (no. 88) were homogeneously isointense to liver in pre- and postcontrast images, but 32 of them exhibited a small high intensity spot in enhanced fat-suppressed images. Malignant lesions and pheochromocytomas (no. 19) had inhomogeneous signal intensity, with relatively higher signal on T2 weighted and Gd-enhanced fat-saturated images. Fat-water images were useful to assess fatty and myeloid components within myelolipomas (no. 7). Benign conditions were distinguished from malignant conditions with 90% sensitivity, 93% specificity and 92.5% diagnostic accuracy. To conclude, modified three-point Dixon technique after contrast agent administration appears to be another useful diagnostic application of midfield MRI to characterize adrenal tissues. PMID- 9221414 TI - [Blood flow assessment with Doppler color ultrasonography in primary and secondary tumors of the liver]. AB - Our study was aimed at measuring hemodynamic changes in liver perfusion in patients with HCC and hepatic metastases using color Doppler US, a noninvasive investigation technique. Eighty-seven patients were examined: 14 of them had HCC and 34 had metastases; the control group consisted of 39 people. Blood flow was measured in the common hepatic artery and portal vein and the ratio of hepatic arterial to total liver blood flow (HPI = hepatic perfusion index) and the ratio of hepatic arterial to portal venous blood flow (A/V ratio) were calculated. HPI and A/V values were changed in HCC patients (HPI = 0.23, range: 0.16-0.35; A/V = 0.32, range: 0.19-0.55) as a consequence of reduced portal venous blood flow (9.76 +/- 2.51 cm3/s) and of increased hepatic arterial flow (2.78 +/- 0.46 cm3/s). HPI and A/V values were significantly changed also in the patients with hepatic metastases (HPI = 0.24, range: 0.11-0.38; A/V = 0.34, range: 0.12-0.61) compared with the control group. These changes were correlated with increased hepatic arterial blood flow (3.16 +/- 1.35 cm3/s) and decreased portal venous blood flow (10.39 +/- 3.81 cm3/s). These results prove the role of color Doppler US in the study of primary liver cancer and metastases. Additional examinations are nevertheless necessary to assess the diagnostic value of color Doppler US in the early detection of and discrimination between benign and malignant tumors. PMID- 9221415 TI - [Instrumental diagnosis of obstructive jaundice: brushing versus biopsy]. AB - We report on a series of 60 patients (31 men and 29 women; mean age: 63 years, range: 37-87 years) submitted to brushing and/or biopsy of the biliary tree through a percutaneous biliary drainage. The tumor involved the biliary bifurcation in 13 cases (21.7%), the biliary duct in 32 cases (53.4%), the right hepatic duct in 1 case (1.6%) and the left hepatic duct in 1 case (1.6%). The metal endoprosthesis was obstructed in 6 cases (10%), while a stenosis of the biliodigestive anastomosis was shown in 7 cases. We used a particular bristle brush and a flexible forceps of the alligator type for cholangioscopic biopsy. Forty-three patients had a malignant stenosis; the diagnosis was confirmed histologically and with clinical follow-up. The comparative adequacy of brushing versus biopsy resulted as follows: 33.3% versus 89.6% sensitivity, 100% specificity for both methods and 59.3% versus 89.6% accuracy. We had 14 versus 3 false negatives and no false positives. Neither brushing nor biopsy brought about any complications. To conclude, biopsy is more accurate and sensitive than brush cytology (91.6% vs 59.3% and 89.6% vs 33.3%, respectively) and therefore it is a safer and faster method to diagnose obstructive jaundice. PMID- 9221416 TI - [Magnetic resonance features in cerebral primary lymphomas in non immunocompromised subjects]. AB - In the past few years, non-Hodgkin's lymphomas have been paid increasing attention to because of their recently increasing frequency. We reviewed the MR images of 17 patients with histologically proved primary CNS lymphoma, all of them immunocompetent at diagnosis. We studied the site, number and shape of the lesions, the presence and grade of edema and possible periventricular spread. The exams were performed with 0.5 T and 1.5 T MR units, using SE sequences on the sagittal, axial and coronal planes, before and after Gd-DTPA administration. The most typical neuroradiologic signs which may suggest the diagnosis of CNS lymphoma are deep or periventricular lesion site, diffuse and marked contrast enhancement, poorly defined borders, moderate edema surrounding the mass and a tendency to periventricular spread. MRI demonstrated 35 lesions in 17 patients. The lymphoma was unifocal in 9 cases (53%) and 7 lesions were localized in subtentorial site. Lesion size did not exceed 2 cm in 49% of cases, ranged 2-4 cm in 40% and exceeded 4 cm in 11% of cases only. These lesions and hypo- to isointense on T1-weighted images (97%) and their signal intensity varies on T2 weighted images, with mainly iso-/hypointense patterns (79%). All lesions enhanced after Gd-DTPA administration, 74% of them markedly and 26% moderately; enhancement was mostly homogeneous (80% of cases). Perilesional edema was observed in 74% of cases. In conclusion, MRI yields some useful information for the diagnosis of primary CNS lymphoma, but the clinical and radiologic signs of this lesion may exhibit aspecific signal features, meaning that no correct diagnosis can be made even in immuno-competent patients. PMID- 9221417 TI - [Role of Doppler color in the differential diagnosis of benign and malignant adenopathies]. AB - The diagnosis of enlarged lymph nodes is of the utmost importance especially in the treatment planning of cancer patients. US yields such morphological findings as node size, longitudinal/transverse diameter ratio, hilum visibility and cortical thickness, which however do not permit the differential diagnosis of benign from malignant forms. Some authors tried to distinguish inflammatory enlargement from metastatic forms on the basis of color Doppler findings, with conflicting and questionable results. We investigated the potentials of color Doppler US in the differential diagnosis of benign and malignant lymph node enlargement using morphological data and flow measurements in lymphatic hilum vessels. The palpable superficial lymph nodes of 70 patients were studied with color Doppler with a linear probe (7.5-10 MHz) equipped for Doppler flow measurements. The largest lymph node was studied in multiple enlargement. The final diagnosis was made with US-guided cytology and/or excisional biopsy. The venous hilar vessels were depicted with color Doppler US in 44/45 patients with lymphadenitis and only in 1/17 patients with metastatic enlargement. Spectral Doppler exams of the hilar arteries showed flows with a wide telediastolic component in lymphadenitis (relative RI:0.58), while flow was rapid and with poor telediastolic component (relative RI:0.84) in metastatic enlargement. Average RI was 0.62 in Hodgkin's lymphomas and 0.71 in all the other lesions. We conclude that the distortion and compression of the main hilar vessels in metastatic lymph node enlargement often prevents color Doppler depiction of venous vessels. Moreover, the compression and distortion of the intranodal capillary network (the "mass" effect) often results in increased RI, as detected with power Doppler in the lymphatic hilum. Even though color Doppler US studies of the hemodynamic changes in the hilar vessels need further validation in larger series of cases, our preliminary results suggest interesting potentials in distinguishing inflammatory from metastatic enlargement, which differentiation remains nevertheless difficult especially in Hodgkin's lymphoma. PMID- 9221418 TI - [Antiblastic locoregional perfusion with control of the aorto-caval flow: technique of percutaneous access]. AB - The authors introduce a new technique for performing aortic stop-flow infusion (ASI) or hypoxic abdominal perfusion (HAP) to treat advanced thoracoabdominal tumors, via an angiographic percutaneous approach. To date, the maneuver has always been performed with surgical exposure of vascular sites in the groins. The materials available on the market were initially used and then dedicated materials have been developed, such as 11-F vascular sheaths, 8-F catheters, latex balloons with maximum phi's of 4 cm and maximum capacity of 70 ml. We performed 72 maneuvers in 56 patients during 22 months. No technical or instrumental complications occurred and all treatments were successful. Three patients (6%) died within 12 hours, two because of ARDS following thoracic perfusion and one for acute renal failure and disseminated intravascular coagulation following abdominal perfusion. The percutaneous approach provides the same mechanical-occlusive efficacy for aortocaval occlusion and therefore the same therapeutic results as surgery, but it has fewer risks of technical complications and no technical failures. Moreover, this technique is more repeatable and less expensive than surgery and its hospital stay and recovery time are shorter. To conclude, the ASI/HAP procedure is an interesting therapeutic chance in otherwise untreatable advanced cancer patients offering several prospects of technical and pharmacologic development to further increase its efficacy. PMID- 9221419 TI - [Assessment of the accuracy of position in breast irradiation with isocentric technique using an electronic system to acquire portal images]. AB - This study investigated the accuracy of daily patient positioning in an isocentric set-up for breast irradiation. This was achieved through the assessment of both systematic and random errors measured as discrepancies between reference simulation images and portal images obtained with an on-line electronic portal imaging device (EPID). To this end, 10 portal images for each of the tangential fields were obtained in 12 consecutive patients and the images were compared to reference simulation films with dedicated software tools provided with the EPID. The discrepancies measured for each set of images were analyzed statistically. Most variations in each series both in terms of cranio-caudal and of lateral displacement appeared to be random, with mean standard deviations of 2.3 and 2.8 mm, respectively. Such variations between reference fields and portal images tend to counteract one another and do not usually play a significant role in the overall accuracy of patient set-up. In some cases, however, the distribution of the variations occurred in a well-defined pattern indicating a systematic error in patient positioning. This study shows that our set-up technique for breast irradiation is relatively accurate and reproducible, and most of the observed errors were below the 5 mm cut-off level, but it stresses the need of accurate quality assurance programs in radiotherapy. Electronic portal imaging devices are excellent tools for fast portal image acquisition, and permit the accurate assessment of discrepancies with respect to reference images. PMID- 9221420 TI - [Brachytherapy with pulsed dosage. General considerations. Radiobiological considerations. First clinical experience in Mestre (Venice)]. AB - The pulsed dose rate (PDR) brachytherapy technique is analyzed and compared with the low and high dose rate (LDR and HDR, respectively) techniques relative to therapy and management, considering the advantages and pitfalls of each technique. From a radiobiological viewpoint, PDR optimization is aimed at obtaining the same therapeutic results as with LDR and HDR relative to both tumor cell killing and possible late damage. PDR permits to administer the same nominal dose rates as with LDR and HDR, but with very different pulse intervals and length. March, 1995, through March, 1996, forty-two patients were treated with microSelectron PDR at the radiotherapy Department of Umberto I Hospital in Mestre (Venice). Twenty-two patients were irradiated on the vaginal vault, 14 on the anal canal, 4 on the breast, one on the endometrium and one on the urethra (the latter patient was a man). Dose rates were 250-300 cGy/h in the vaginal vault and 90 cGy/h in the other sites. Source-dwell interval in the applicators was 2.5 mm, dwelling time for each position ranged 6.8-122 s, 3-73 pulses were applied lasting 167-1958 s. The unit was reliable and the only problem was the need to recalibrate it every 5-6 applications because of computer memory saturation. Because of the short minimum follow-up (3 months), only the early reactions to treatment have been assessed: no toxicity was found in the vaginal vault, endometrium and breast. Low-grade proctitis was observed in 11 of 14 treated anal canals and another patient complained of more severe symptoms for two weeks; the disease progressed in two anal canal patients, as in the urethra patient. In conclusion, PDR brachytherapy appears a reliable technique whose early clinical results are encouraging. PMID- 9221421 TI - [Management control and operative outcome of the Radio Institute "O. Alberti." Management of oncologic follow-up]. AB - Post-treatment cancer patient surveillance is an area with few given standards where the need of guidelines has become imperative with the recent emphasis on controlling the ever-increasing health care cost. Unfortunately, literature reports are often inconclusive and ambiguous, mostly because of the lack of properly controlled trials comparing the cost and benefits of various follow-up protocols. In addition, the actual impact on patient survival and quality of life is questionable. At the Istituto del Radio "O. Alberti" (IRA), we consider the follow-up as a sort of population screening aimed at the early detection and treatment of recurrent disease. While aggressive surveillance undoubtedly detects some cancers before symptoms develop, it is debated whether the impact on survival and quality of life are measurable. The early detection of relapse is only a potential survival benefit if recurrent disease is curable with further treatment or at least if salvage treatment is more effective in patients with less severe disease. We investigated the effectiveness, efficacy and medical care of our follow-up protocol. April to June, 1996, we examined 1,223 of 2,148 expected patients; 225 patients disattended the scheduled visits. IRA spent about It. L. 33,800 per examination. Fifty-seven patients were hospitalized to carry out treatment and IRA hospitalization charges were about It. L. 1,100,000 while overall social expenses were about It. L. 6,600,000. Regular visits to see an oncologist provide easy access to specialist medicine and convey a sense of being looked after with a caring system. 94.5% of patients prefers to continue the follow-up program with scheduled visits. Most patients (70%) know about the examinations they undergo but consistently overestimate the importance of laboratory tests and imaging findings and underestimate the importance of medical history and physical examination. In addition, most patients (95%) misinterpret the term "normal" relative to a test result. This study suggests that patients are unfamiliar with the limitations of more costly diagnostic and follow-up studies, which reflects the fact that physicians spend little time discussing follow-up strategies with their patients, especially regarding the cost-benefit analysis and the sensitivity and specificity of the laboratory tests and imaging examinations. Finally, in our opinion follow-up cost is acceptable even though expenses should be reduced optimizing the request of instrumental examinations. Therefore, oncologists should definitely try to inform their patients about the clinical importance of follow-up. PMID- 9221422 TI - [Analysis and comparison of various quality protocols for radiotherapy linear accelerators]. AB - The main parameters determining the quality of an electron beam produced by a linear accelerator for medical use were considered in this study, particularly: flatness, symmetry and uniformity. We analyzed and compared several protocols issued by national and international associations (such as the AAPM, IPSM, ICRU, NACP), the software protocol developed for the measurement system we used (Multidata) and the measurement instructions recommended by the accelerator manufacturer (Siemens). The above associations issue quality protocols to ensure system performances suitable for medical use, to increase patient safety and to improve the treatment outcome. Radiation therapy safety and improvement depend on correct dose measurements and dose distribution in the treated volume. Once the dose value per monitor unit ratio (Gy/M.U.) is determined, controls are necessary to be sure that the value does not change in time and that the dose distribution has the same effect in the whole treated volume. Our goal is to point out the differences and the affinities in the definition of the parameters, which change slightly in the different protocols, and to study the origin of the differences found when the experimental results were compared. Another important issue is represented by the frequency of quality controls, which are definitely different from the fast checks which are often performed. In conclusion, some suggestions are provided for the choice of the quality protocol to follow. PMID- 9221423 TI - [Quality evaluation of a personal dosimetry service]. AB - In a hospital environment high quality personal dosimetry is demanded by two different considerations: first, the marked reduction in the radiation exposure levels of hospital workers during the last 10 years and second, the recent decrease in the allowed absorbed dose thresholds for the different categories of workers and for the general population. In fact, according to the new Italian Radioprotection Law (D.L. vo 230/95), the dose equivalent limit for the general population has been decreased to 1 mSv per year. This means that a dosimetric system should be able to measure, with acceptable precision and accuracy, dose levels as low as 0.1 mSv per observation period (generally 1 month or 45 days). This is quite a stringent requirement for this kind of dosimetry. During a tender, the performances of the whole body personal dosimetry systems by four Italian service providers were analyzed by irradiating more than 60 test samples for each provider with four different energies in a wide dose interval (0.01-100 mSv). The results show that all systems perform quite well in the 0.2-100 mSv dose range; on the contrary, in the 0.01-0.2 mSv dose range, significant differences appear between the services and TLD based systems perform better than film based ones. In particular, one of the two TLD based systems measured doses as low as 0.01 mSv. To conclude this very high sensitivity level really opens a new "observation window" on the low doses world. The use of higher quality (and, of course, more expensive) materials by this provider seems to be the key of its success. PMID- 9221424 TI - [Malpractice claims against radiologists in Italy. Trends in 1993-1995]. AB - The insurance claims against Italian radiologists over a three-year period (1993 1995) were anonymously reviewed, based on pertinent data provided by the Insurance Company of the Italian Society of Medical Radiology. The incidence risk rate of claims was 11.4 per thousand persons/year. The overall claims rate increased in 1995. Alleged malpractice accounted for more than 85% of the claims. Misdiagnosis represented the first and most important claim category (43.4% of the total). The most common misdiagnosis was the failure to diagnose fracture or dislocation. The second most common plaintiff's misdiagnosis argument was the failure to diagnose breast cancer. The second most frequent claim category (35.8%) were complications, frequently occurring during interventional radiology and contrast media injection. A minority of claims (11.3%) originated from patient injury occurring in the radiology department during exam execution. Finally, radiologists were frequently sued together with medical (or surgical) doctors in case of patient death, according to an Italian law (Art. 589 P.C.). Claims were more frequent in public health services and they were mostly related to emergency examinations and interventional procedures. PMID- 9221425 TI - [Magnetic resonance findings in a case of intraosseus fibular lipoma]. PMID- 9221426 TI - [Magnetic resonance contribution in a case of multiple tenosynovitis of the ankle caused by orthopedic shoe]. PMID- 9221427 TI - [Muscle metastasis in a patient with epidermoid carcinoma of the lung]. PMID- 9221428 TI - [Extraosseus osteogenic sarcoma: report of a case with atypical secondary involvement of the lung]. PMID- 9221430 TI - [Unusual etiology of Budd-Chiari syndrome: pericardial cyst]. PMID- 9221432 TI - [Boerhaave syndrome: report of 2 cases]. PMID- 9221429 TI - [Imaging with magnetic resonance in a case of post-traumatic aneurysm of the radial artery]. PMID- 9221431 TI - [A case of duodenal duplication causing invagination in a patient with intestinal malformation: non-invasive imaging]. PMID- 9221433 TI - [Tuberculous pancreatic abscess in a case of miliary lung tuberculosis]. PMID- 9221434 TI - [Rendu-Osler-Weber disease: embolization of pulmonary arteriovenous fistulas with tungsten coils. Technique and results in 3 cases]. PMID- 9221435 TI - [Personal identification with ante- and post-mortem computerized tomography. Report of a case]. PMID- 9221436 TI - [Old, glorious traditional radiology ...]. PMID- 9221437 TI - [Ulcerative colitis and dyspnea]. PMID- 9221438 TI - [Liver metastasis and nuclear medicine]. PMID- 9221439 TI - Proceedings of the 1st Latin American Congress on Acute Renal Failure. Sao Paulo, Brazil, March 27-29, 1996. PMID- 9221440 TI - [Children from the GAZEL cohort: II--motive for contact with the medical educational system by age and sex]. AB - An epidemiological survey of 2582 French children aged 4 to 16 has been conducted to assess patterns of service use in relation to psychological disturbances. Details on the design, sample, survey instruments, response rate, and 12-months prevalence rates of contacts with a range of different professionals were presented in a previous article. In this second article, the psychological motives leading to contact with family doctors, school-based professionals, speech and language therapists, and mental health specialists are analyzed. The age and gender effects are assessed for each motives. On the whole, consistent sex differences were found for the types of complaints presented by service users, with emotional symptoms being more frequent amongst girls, and behavioural, developmental and learning difficulties being more frequent amongst boys. Mental health specialists were attended for a variety of reasons. Family doctors were contacted for minor emotional difficulties. Because family doctors were consulted by a high proportion of children and adolescents of our sample, the role of this professionals in the detection and management of minor psychological morbidity is emphasized. PMID- 9221441 TI - [Modelling of length of stay and costs in 2 homogeneous groups of hematological and pneumological patients: clinical characterization of patients with long-stay and high costs]. AB - After the implementation of the Medicare Prospective Payment System (PPS) in the USA, many European countries like France have introduced DRGs to curb hospital overspending. However, there has been some reluctance from hospital actors, especially because of the heterogeneous nature of DRG's. To analyse this situation, we propose a method based on distribution modelization of length of stays and costs within DRGs. For each DRG, the model is based on a mixture of Poisson and Weibull distributions identified as subgroups. The subgroups are characterized by their means and their proportions which are estimated by maximization of data likelihood. For a particular DRG, the proportion of long stay or high-cost patients can be explained by the introduction of clinical variables in the model. First the model was applied to the DRG "leukemia and lymphoma" (HCFA V.3), using 133 discharge abstract files from the Dijon public teaching hospital which were classified into this DRG in 1993. Among the studies parameters only acute leukemia, neutropenia < 500 PNN/mm3, high dose aplastic chemotherapy, central venous catheterization, parenteral nutrition, use of protected or laminar air flow room, septicemia, large spectrum intravenous antibiotherapy, and blood transfusion had a significant influence on the distribution of the patients in the long stay or costly subgroup. Second, for DRG "chronic bronchopneumopathies" (n = 220) the significant parameters were mechanical ventilation, antibiotherapy, post hospitalization medicalized care. PMID- 9221443 TI - [Empirical Bayes estimates of relative risk: principles and examples]. AB - Most atlases, such as cancer incidence or mortality atlases, show information for geographical subunits with major differences in population size. Under such conditions, classical epidemiological mapping indexes (SIR, SMR) are unsatisfactory. Regional pattern, if they exist, cannot be shown. Empirical Bayes methods produce smoothed relative risk estimators useful for solving this problem. We present some elementary principles of these methods using cancer incidence data in the French department of Isere. PMID- 9221442 TI - [Cost-effective approach to the screening of HIV, HBV, HCV, HTLV in blood donors in France]. AB - In order to provide greater safety in blood transfusions, public health authorities have imposed the use of screening tests. The purpose of this paper is to estimate the cost-effectiveness ratios of the screening test used in France. Four risks were studied: HIV, HBV, HCV and HTLV. Two efficiency measures were used: cost per positive blood donation detected and cost per case of prevented infection transmission. Moreover, for HTLV alone, the efficiency was estimated by the cost per prevented pathology. Data concerning the costs were provided by the French Blood Agency; those concerning the results of the screening campaigns were provided by the official health authorities, the other data used in the calculations were drawn either from the French Blood Agency data or from a review of international literature. Results gave information about the expenditure devoted to the screening of virologic risks associated with blood transfusion in France (250 million francs per year for the four viruses studied). They stressed the differences in screening efficiency according to the test studied (the cost by prevented seroconversion varied from 31,795 francs for HBV, 72,180 francs for HCV, 676,596 francs for HIV to 6,137,346 francs for HTLV screening test in the base case) and especially the very low efficiency of the systematic screening of the HTLV virus (from 34 to 307 million francs per prevented leukemia). PMID- 9221444 TI - [Estimation methods in a sampling survey]. AB - The objective of this paper is to present a guide for statistical analysis of a sampling survey. Advantages and disadvantages of classical sampling designs are first discussed. A sampling survey was designed to give more precise estimates of parameters which characterize a targeted well-defined population. Simple sampling design often is impossible in large population and rarely is the optimal solution. According to practical and economic constraints, and to available sampling frames, other strategies (stratification, unequal probabilities, several selection steps) may be necessary or more efficient. Fundamental tools to compute estimators and their confidence intervals are presented. The choice of the sampling method determine for each population unit the probability to include it in the final sample. These inclusion probabilities must be known to formulate estimators, ideally unbiased and with low variance. Difficulties arise from calculating the variance, especially for estimators which are not linear functions of the characteristics of interest. In that case, estimation procedures include Taylor linearization. It is always possible to find at least one linear estimator for a total. The total is the key-parameter in sampling theory, most parameters (such as ratios, means, percentages...), being function of unknown totals. Numerical examples are given. PMID- 9221445 TI - [Tuberculin test, antitubercular chemoprophylaxis and incidence of tuberculosis in a Swiss HIV cohort study]. PMID- 9221446 TI - [Maternal mortality in the National Hospital Center of Ouagadougou (Burkina Faso). Report of 123 cases collected in 1995]. PMID- 9221447 TI - [Mass screening for colorectal cancer using occult blood studies. From theoretic efficacy to generalized practice]. PMID- 9221449 TI - [Vaccines and vaccination]. PMID- 9221448 TI - [4 studies on home birth]. PMID- 9221450 TI - [Survival after 60 years of age among elderly retired miners]. AB - This study analyses long term effects of working conditions at an advanced age and post retirement. A longitudinal study was done among 63,000 retired miners between 60 to 65 years old. The standardised mortality ratio (SMR) in the cohort was above that expected on the basis of concurrent mortality in the reference population (SMR = 111; 95% confidence interval (CI): 108-114). The excess mortality was observed in occupational groups with exposure to underground tasks (10 years of underground tasks: RR = 2; 95% CI: .1.7-2.5), but there was sign of a healthy worker effect after 25 underground years. The effects of modern technologies and of medical progress have changed working conditions and care has reduced mortality for retired manual workers: exposure to underground tasks before 1950: RR = 1.7; CI 95%: 1.4-2.0; after 1950: RR = 1.2; CI 95%: 1.1-1.2. PMID- 9221451 TI - [The stent in cardiologic interventions. Introduction]. PMID- 9221452 TI - [The effectiveness of stents in restinotic lesions and coronary grafts]. AB - Regarding restenotic lesions, the data suggest that stent implantation decrease the number of cardiac events including the restenosis rate. The higher number of acute cardiac events are related to stent thrombosis, and they are markedly reduced by the new techniques of antiagregation and implantation. The restenosis rate post-stent implantation does not bear relation to the number of previous dilations as it does with conventional angioplasty. Regarding vein grafts, the implantation of different types of stents has a high percentage of success and low rate of acute complications. The longterm survival of these patients is related more to the progression of the coronary arteriosclerosis and tend to worsen with time. Although the data on restenosis are better than with conventional angioplasty, we need to wait for the definitive results after the final conclusions of the randomized trial SAVED. PMID- 9221454 TI - [Stents and de novo coronary lesions. Meta-analysis]. AB - OBJECTIVES: The purpose of this investigation was to provide an overview of the effect of coronary artery stent and balloon angioplasty on major clinical outcomes and restenosis from 3 trials (BENESTENT, STRESS and START) comparing the two treatments in the novo lesions. METHODS: The trials included a total of 1,374 patients, 693 of whom were randomized to stent and 682 to conventional angioplasty. Relative risk, 95% confidence interval and p values were calculated for death, infarction and need for revascularization at 6 month follow-up. Restenosis was assessed by using the binary definition. RESULTS: The incidence of combined clinical outcomes was similar to the two treatment strategies (relative risk 0.96; 95% CI 0.64-1.45). However, the odds for needing a new revascularization procedure were reduced by 35% in patients treated with stents (relative risk 0.65; 95% CI 0.51-0.82; p < 0.001). Restenosis rates were 25% and 36.6% (stent vs angioplasty, respectively), which represents a reduction of 31% (relative risk 0.69; 95% CI 0.58-0.81; p > 0.0001). CONCLUSIONS: Compared with balloon angioplasty, coronary stents of de novo lesions in native coronary arteries decreases restenosis at 6 months. This translates into clinical benefit, as shown by a reduction in the number of new revascularization procedures. PMID- 9221453 TI - [Intracoronary stents in the treatment of angioplasty complications]. AB - Acute or subacute occlusion of the dilated artery is the main cause of complication after percutaneous transluminal coronary angioplasty. The occlusion incidence ranges between 2 and 10% of the procedures and the mechanisms include dissection, subintimal hemorrhage, thrombosis, vasospasm and elastic recoil. Intracoronary stent is a metallic support which may seal dissection due to angioplasty, resists elastic recoil and geometric remodeling and prevents other occlusion mechanisms. Data from recent studies demonstrate the benefit of stenting acute occlusive coronary dissection in a large artery with extensive myocardium at risk. The election of the type of stent in the treatment of the acute coronary occlusion depends on the clinical status, the coronary anatomy and the stent characteristics. The major limitation of stent is the apparition of subacute thrombosis. The mechanisms of stent thrombosis are not well known. However, some factors related to complicated angioplasty, such as the presence of important dissection, intracoronary thrombus and vessels of small diameter have been involved. For this reason, the major incidence of subacute thrombosis is observed in bail-out situations. New techniques using intravascular ultrasound and high pressure dilatation after stent placement and new approaches in antiplatelet treatment have been recently described in order to reduce the incidence of subacute occlusion and the hemorrhagic events associated to previous anticoagulant therapy. PMID- 9221456 TI - [Is there one type of stent for every lesion?]. AB - Intracoronary stents are, without any doubt, a major breakthrough in interventional cardiology. Their widespread use has expanded to more difficult anatomical situations and the search for more suitable stents continues to grow. We review, in this paper, the technical characteristics of stents that are currently approved or in clinical investigation. We have also reviewed the role of intravascular ultrasound in the study of the anatomical characteristics of plaque, the length of the lesion, and their influence of the stent selection and the ultrasound influence on the determination of appropriate stent expansion. After reviewing the current role of the intracoronary stent, we tried to look for the most appropriate stent in three "unconventional" anatomical situations: long and bifurcated lesions, lesions containing thrombus and saphenous vein aortocoronary bypass grafts. In conclusion, the intracoronary stent plays a major role in interventional cardiology. The second and third generation stents are more suitable for "specially difficult" situations, but there are some lesions such as bifurcations where the is not yet a definitive solution. PMID- 9221455 TI - [Post-implant antithrombotic treatment after intracoronary stents. Thrombotic occlusion]. AB - The stent has been demonstrated to be a useful device in the treatment of complicated coronary angioplasty and in the prevention of restenosis. However, its efficacy was seen to be initially limited due to a high incidence of thrombotic occlusion of the stent in the first month after implantation and a high rate of hemorrhagic complications when a severe antithrombotic treatment with antiplatelet drugs (aspirin and dypiridomole) was associated with anticoagulation therapy with intravenous sodium heparin and dicumarol. Both phenomena increased morbidity and the post-implantation costs of stenting. The development of new strategies in stent implantation and post-implantation management have significantly reduce these complications. The objective of this study is to review the physiopathology of thrombotic occlusion following intracoronary stent implantation and the efficacy of various antithrombotic pharmacological strategies being used for its prevention. Although certain factors existing prior to implantation (thrombus, severe dissection, and the size of the vessel) augment the probability of occlusion in the stent, the result of the implantations is a good predictor of the development of this complication. Recent studies have shown that when optimal coronary stent implantation (high pressure, strict angiographic or ultrasound criteria) resulting in a minimal or absent residual stenosis and adequate apposition of the stent against the arterial wall is associated with new antithrombotic strategies, the rate of thrombotic occlusion should be less than 1.5% and the rate or hemorrhagic complications should not be greater than what has been described for conventional angioplasty. The most consolidated current antithrombotic therapy is the association of aspirin and ticlopodine which has demonstrated its efficacy in both observational and randomized studies. The combination of antiplatelet drugs and low molecular weight heparin has also demonstrated its efficacy in non randomized studies and may constitute an alternative in some clinical or angiographic situations. The development of stents with a smaller thrombogenic surface contact with blood (made of materials which are not thrombogenic or are coated) hopefully provides another possibility for the near future. All of these advances have minimized the problem of thrombotic occlusion of the stent and have contributed to the great expansion in the use of this technique in current interventional cardiology. PMID- 9221457 TI - [The role of non-angiographic observations (IVUS, angioscopy, doppler) in coronary stenting]. AB - Coronary stenting is increasingly used during transcatheter coronary therapy. Coronary angiography, mainly since the advent of quantitative angiography, provides an effective tool to obtain excellent clinical results with coronary stenting. However, during this procedure some limitations inherent to the angiographic techniques may become apparent. Accordingly, great enthusiasm has been generated regarding the potential value of alternative diagnostic techniques to guide coronary stenting. Intravascular ultrasound is able to study the arterial wall and provides a unique tool to assess stent expansion, apposition and symmetry. Therefore, this technique is increasingly used to optimize stent deployment. Coronary angioscopy directly visualizes stent expansion and is able to precisely recognize protrusion of redundant fronds of tissue or residual dissections within the stent struts. In addition, this technique is the procedure of choice to identify intracoronary thrombus. Intracoronary Doppler permits the application in the catheterization laboratory of sophisticated methods of functional assessment of lesion severity. Coronary stenting allows a faster and complete normalization of coronary flow reserve than balloon angioplasty. Thus, all these new techniques of intracoronary diagnosis provide unique and useful information, which is complementary to that obtained with angiography, potentially useful during coronary stenting. PMID- 9221458 TI - [Primary stent treatment in the acute phase of myocardial infaction]. AB - INTRODUCTION: Although direct balloon angioplasty has emerged as an alternative to thrombolytic therapy in patients with acute myocardial infarction, reocclusion and restenosis rates are limiting factors. We postulated that these limitations could be partly overcome by primary stenting of the responsible lesion. MATERIAL AND METHODS: Since January/94 we have studied 59 patients with acute myocardial infarction who were treated in the early phase (3.1 +/- 2 hours since the onset of symptoms) by elective Palmaz-Schatz stent implantation. No adjunctive thrombolytic therapy was associated. Two patients were in cardiogenic shock and were treated under percutaneous cardiopulmonary support. At cardiac catheterization a left ventriculography and coronary angiograms were obtained. Then, mechanical recanalization of the responsible lesion was performed. If the angiographic anatomy was considered suitable, a stent was implanted at the lesion. RESULTS: The infarct related artery was the left anterior descending in 29 patients, the circumflex in 14 and the right coronary artery in 16. At baseline conditions, 40 patients had a totally occluded artery and 19 showed a TIMI-grande 1 antegrade flow. One patient had an early clinical recurrence 4 days later, which required an additional divided Palmaz-Schatz stent at the distal portion of the lesion, in order to seal a residual dissection. All remaining patients had a favourable clinical course without major complications. Immediately after treatment the minimal lumen diameter was 3.2 +/- 0.4 mm and no residual stenosis was detectable at the treated segment. Six-month angiographic reevaluation was performed in all 29 (49%) eligible patients. Restenosis (> 50% stenosis) was detected in 6 out of the 29 evaluated patients (21%). CONCLUSIONS: Primary stent implantation in selected patients with an evolving myocardial infarction provides good initial and 6-month results. PMID- 9221459 TI - [The role of the stent in non-coronary cardiopathies]. AB - INTRODUCTION: The stent has demonstrated to be a useful device in the prevention of postangioplasty coronary restenosis and it is expected to have a favourable effect as an alternative or complementary treatment of stenotic lesions in arteries or veins associated with congenital defects. The aim of this study is to analyze our experience in this setting. MATERIAL AND METHODS: From February 1992 to March 1996, 28 stenting procedures were performed in 26 patients (mean age: 8.6 +/- 0.7 years; mean weight: 26.2 +/- 3 kg). In 12 patients, stenting was single, and a iliac Palmaz stents were always used. Stenting location was: pulmonary artery branches in 17 patients, right ventricular outflow in 2 patients, in the junction of right atrium with pulmonary artery in 2 patients, systemic veins in 2 patients and in post Mustard intratrial channel stenosis in 2 patients. 25 patients had previously undergone at least one surgical procedure. RESULTS: The stenotic diameter of the treated lesions increased significantly after the procedure (4.4 +/- 0.3 mm before stenting vs 11.6 +/- 0.3 mm after stenting, p < 0.0001) and the transtenotic gradient decreased from 38.1 +/- 5 to 12 +/- 3.8 mmHg. Those changes were associated with a diminution of right ventricular pressure (81.6 +/- 3 vs 56.7 +/- 6 mmHg, p < 0.0001) in patients with pulmonary branch stenosis without septal defects. There was no mortality among the percutaneously treated patients and only one patient needed surgery. Nevertheless, one patient died after bilateral intraoperative stenting. CONCLUSION: The treatment of proximal or distal stenotic lesions in the pulmonary tree, systemic veins, and obstructed intraatrial channels with stents, can replace or complement conventional balloon angioplasty. It also offers a useful and effective alternative to surgery, when it is impossible or carries a risk. PMID- 9221460 TI - [Intracoronary prosthesis (stent): an approach to the analysis of cost effectiveness]. AB - Stents have emerged as one of the major therapeutic tools for percutaneous intracoronary revascularization procedures. In fact, a stent is implanted in at least 30% of lesions attempted. Their clinical impact is huge because stenting has produced a decrease int the need for emergency surgery to 0.5%, with an incidence of acute myocardial infarction related to angioplasty of 2% and a death rate of < 1% despite unfavourable clinical and anatomical conditions treated. The initial price of stenting was a high frequency of subacute stent thrombosis and peripheral vascular complications, which has been solved. In this context, the cost of stenting procedures increased by more than 30% in relation to the cost of conventional balloon angioplasty. But, the use of new antiplatelet regimes, implantation with high atmospheres and the achievement of a minimal residual narrowing after stenting practically promotes the disappearance of the two major initial problems, making possible a decrease in the restenosis rate to 20% with a parallel reduction of 10% in the need for new revascularization procedures during the first years after stent implantation. This means a reduction in the midterm costs. The incremental rate of cost/effectiveness after stenting in one vessel disease has been estimated as 23,600 dollars/years of adjusted quality of life gained ($/QUALY) ($20-40,000/QALY is acceptable in pharmaeconomic studies). From another point of view, stenting implies 19 QALYS gained at a mean cost of $52,700. The incremental cost of stenting may be more favourable if a reduction of $1,800 in the total cost of stenting procedures could be achieved, or if the stent cost were reduced by at least $700. In Spain, each patient who was event free during the first year of follow-up would cost 1,674,000 ptas. A 10% reduction of new revascularization procedures x 100 patients would cost 20 million pesetas. Despite the enormous interest of socioeconomic analysis these data only reflect a partial point of view, because of the complexity of evaluating the contribution of new technologies, the true quality of life gained for patients, the societal point of view, or that of the National Health Service. Moreover, we assume that in the current socioeconomic context there are finite resources and a limitless demand. PMID- 9221461 TI - [Future perspectives in coronary stenting]. AB - Coronary stenting has made a significant difference in percutaneous coronary revascularization techniques, since it provides an effective treatment for procedural complications and prevents to some degree the incidence of restenosis. Presently, many different stents are available on the market, therefore most interventional cardiologist needs are well covered. Nevertheless, there are still some vessels/lesions unsuitable for coronary stenting, such as very distal lesions or lesions in small vessels. On the other hand, although restenosis is less frequent after stenting, its incidence is still significant posteriorly, representing a major health and economical problem. In the near future, new developments in stent technology such as polymeric coating, local drug delivery systems or intraarterial radiation may contribute to a further decrease in the incidence of restenosis. With better short- and long-term results indications for percutaneous coronary revascularization might be significantly expanded in the next few years. PMID- 9221462 TI - [Digestive and extra-digestive complications of nonsteroidal anti-inflammatory drugs. Preventive and curative strategies]. AB - The authors review the digestive ulceration risk factors and the criteria for selecting a non steroidal antiinflammatory (NSAI), included the most recent drugs, such as selective anti-cyclo-oxygenases 2. They actualize the preventive strategies and insist on the values of misoprostol and of slow acting anti rheumatic drugs. In the case of digestive ulcerations, they plead for the immediate stop of the NSAI and its replacement if necessary by corticosteroids, for the prescription of a proton pump inhibitor (PPI) or mesalazine according to the localisation of the lesion, finally for the eradication within 8 days of Helicobacter pylori. PMID- 9221463 TI - [Glycosylated hemoglobin in nonselected under-18-year-old young diabetics. Comparison of 2 HPLC methods]. AB - The aim of this study is to determine, in an unselected population of diabetic children and adolescents < 18 years of age, which HbA1c levels can be achieved, and to compare 2 different HPLC (High Performance Liquid Chromatography) methods. One hundred and fourty-four unselected subjects (73 boys and 71 girls), with a diabetes duration ranging from 5 months to 15 years were included. HbA1c was measured simultaneously in our hospital and at the Steno Institute in Danemark. The mean (SD) HbA1c levels in the 144 children and adolescents were 6.6 +/- 1.2% using our method (normal values: 3.9-5.5%), and 7.7 +/- 1.1% at the Steno Institute (normal values : 4.4-6.3%), with a mean difference of 1.1 +/- 0.4%. Therefore, one must be cautious when comparing HbA1c levels, even by HPLC, between laboratories. Moreover, the lack of HbA1c standardization makes difficult to define HbA1c thresholds in order to avoid diabetic complications. In more than 60% of the patients, it is possible to obtain an HbA1c level under the normal mean value plus 5 SD. In the other paediatric studies, HbA1c levels are higher, whatever the type of treatment. PMID- 9221464 TI - [Sleep apnea and nocturnal ventilatory assistance (nCPAP): 5-year experience in the conventional system]. AB - The reimbursement of nasal continuous positive airway pressure (nCPAP) by the Belgian social security, via a conventional system, has made since 1991 this treatment available to an increasing number of patients having moderate to severe sleep apnoea hypopnoea syndrome (SAHS). We have reviewed our experience in prescribing domiciliary nCPAP from 1991 to 1995. Three hundred twenty-five subjects with SAHS, predominantly male (89%) and/or obese (77%) subjects, have benefited. Mean use of nCPAP machine, assessed by reading the time counter, amounted 4.7 h per 24 h, with only 23% of non-compliant patients (use < 3 h per 24 h). In 205 patients nCPAP was effective in controlling SAHS-related symptoms. Cure, with successful weaning from nCPAP, was obtained in 16 patients, as a result of marked weight loss in 13 of them. Forty-six non-compliant subjects were not allowed by the physician to go on, and 40 subjects left nCPAP because of intolerance. Finally, 10 patients abandoned nCPAP because of inefficacy, ascribed to some associated condition, being predominant, and 8 patients died. Our results suggest that domiciliary nCPAP is an effective treatment for SAHS in a majority of subjects, but that this kind of treatment is prescribed lifelong, unless there is a marked weight loss. The Belgian conventional system, as it requires a regular follow-up, contributes to keep non-compliance within acceptable limits. PMID- 9221465 TI - [Neonatal screening for hemoglobinopathies in the Brussels region]. AB - We realised this study in order to determine the frequency of abnormal haemoglobins and to appreciate the need for a neonatal screening for haemoglobinopathies in Brussels. Over a two year-period, 9575 cord blood samples were systematically screened. The study disclosed following results : 40% of newborns were from regions at risk for haemoglobinopathies and abnormal haemoglobins were present in 2.5% of the neonates tested. This frequency is similar to those reported elsewhere in North Europe. The most frequent abnormal haemoglobins were the Hb S, Bart's, C, D and E. Three cases of severe forms of sickle cell anaemia were identified. The frequency of abnormal haemoglobins and Hb S traits combined to the high rate of mixed marriages (16%) justifies the need for a universal screening for haemoglobinopathies in Brussels. PMID- 9221466 TI - [Anatomoclinical conference: breast cancer, hyperleukocytosis and respiratory failure]. PMID- 9221467 TI - [Importance of current methods in interventional cardiology and heart surgery]. PMID- 9221468 TI - [Measurements of intracoronary pressure and blood flow velocity]. AB - Quantitative coronary angiography and new intracoronary imaging devices are not able to provide functional data for the assessment of the severity of coronary artery stenotic lesions. The functional characterization of coronary artery stenosis gives insight into whether it compromises myocardial perfusion under conditions of pharmacologic or physical stress, i.e., whether the stenosis has hemodynamic relevance. The measurement of trans-stenotic pressure gradients and/or post-stenotic blood flow velocities using very recently developed, miniaturized pressure- and Doppler-angioplasty guidewires (1/3 mm in diameter) provides a valuable alternative to traditional, non-invasive means for the functional assessment of coronary artery disease. The most widely employed parameters for the functional characterization of coronary artery stenoses are the ratio between flow velocities during pharmacologically induced hyperemia and at resting conditions (coronary flow velocity reserve), and the pressure-derived fractional flow reserve, i.e., the ratio of mean poststenotic to mean aortic pressure during hyperemia. Furthermore, poststenotic pressure and flow velocity measurements during and after occlusion of stenosis can be used for the quantitative assessment of collateral circulation among different vascular regions. PMID- 9221469 TI - [Use and significance of intravascular ultrasonic imaging]. AB - Intravascular ultrasound (IVUS) imaging of coronary arteries has recently become possible in vivo with the improvements achieved in the miniaturisation of ultrasound transducers. IVUS provides informations complementary to angiography, and is considered as the gold standard for the assessment of lumen size, plaque thickness and plaque distribution. As formerly demonstrated by pathological studies, IVUS also reveals an underestimation of plaque thickness with angiography and an incomplete assessment of the true morphologic distribution of the plaque burden. Among its clinical applications, IVUS can be used to monitor revascularisation procedures, and may provide very accurate measurements of progression or regression of coronary atherosclerosis and of the extent of posttransplant vasculopathy. IVUS was employed successfully to assess and optimize the results of percutaneous transluminal coronary angioplasty. Clinical studies have shown that the risk of restenosis is inversely proportional to the size of the postprocedural lumen. With the guidance of percutaneous transluminal angioplasty by IVUS, balloon size and inflation pressures were increased, and better results were obtained with larger lumen size in comparison to the results of the procedures assessed by angiography alone. These improvements contributed to abandon the anticoagulation after stent implantation in most cases. In spite of the valuable information provided by IVUS, its role in clinical settings should still be defined IVUS prolongs the revascularisation procedure and enhances its costs and risks. Thus, the cost effectiveness of IVUS in its clinical application should be determined by prospective studies. PMID- 9221470 TI - [Current trends in coronary angioplasty]. AB - Coronary angioplasty still is an area of intensive development. The publication of the BARI data confirms and consolidates previous findings that balloon angioplasty and bypass surgery are equivalent in terms of mortality and major complications over a mid-term follow-up period. However, balloon angioplasty has a high rate of reinterventions, and is not recommended in diabetic patients with multivessel disease. The question, whether direct PTCA is a better treatment strategy of acute myocardial infarction than thrombolytic therapy remains unsettled despite a slight advantage for direct PTCA in several small randomized trials. The role of coronary stents has continuously expanded over the last years, as a tool against acute as well as long-term complications of angioplasty. This success is partly due to the finding that antiplatelet agents such as ticlopidine offer an effective protection against subacute stent thrombosis. The introduction of the GP IIb/IIIa platelet receptor blockers has provided another powerful tool against acute thrombotic complications, leading to a more aggressive interventional strategy in acute coronary syndromes. The development of alternative "devices" has further slowed down because of repeated publication of negative clinical results. Although different forms of laser energy applications, of local drug delivery, and of local irradiation are interesting research areas, their current impact on clinical practice is small. PMID- 9221472 TI - [Diagnosis of diabetes mellitus: oral glucose tolerance test or HbA1C]. PMID- 9221471 TI - [Interventional treatment of heart valve diseases]. AB - Percutaneous balloon valvuloplasty of stenoses has been introduced into medical practice in the late 70ies. Over the past decade, the method has evolved to a valid alternative to valve surgery in selected cases. Balloon valvuloplasty of isolated mitral stenosis is to date the therapy of choice and yields results comparable to those of surgery. It is even superior in only moderately diseased valves. However, the most frequent valve stenosis, that is aortic stenosis of the elderly, is not suitable for balloon dilatation. Dilatation of congenital aortic stenosis can be attempted if the valve is bicuspid or tricuspid. The recurrence rates for valvular stenoses after valvuloplasty are similar to those after surgical commissurotomy. PMID- 9221473 TI - [Inflammation of the lower leg]. PMID- 9221475 TI - [Mechanism of HIV infection]. PMID- 9221474 TI - [Atrial septum defect and open foramen ovale. Diagnosis, clinical aspects and therapy using a catheter technique]. AB - The purpose of this report is to describe the pathophysiology and clinical symptoms of atrial septal defects with emphasis on right ventricular failure and the risk of paradoxical embolism in the setting of a patent foramen ovale. The diagnosis of both the atrial septal defects and the patent foramen ovale, usually is made by echocardiography. Percutaneous transcatheter occlusion of an atrial septal defect and a patent foramen ovale has gained interest for some years as a treatment alternative to surgical closure. Preliminary results of different devices have been encouraging and the method appears to be a viable alternative to surgical closure. PMID- 9221476 TI - [Interventional cardiology in pediatrics]. AB - Since the first description of successful pulmonary balloon valvuloplasty in 1982, the field of pediatric interventional cardiology has seen a dramatic development Nowadays, interventional techniques are a standard therapy for a wide range of indications in congenital heart disease: balloon pulmonary and aortic valvuloplasty are therapies of choice as well as the occlusion of the persistent ductus arteriosus by double umbrellas or coils. Dilatation of a recoarctation after initial surgical therapy is safe and successful whereas there is an ongoing debate whether native coarctation should be managed surgically or by catheter intervention. Other common indications for catheter interventions are dilatation and stenting of native or postoperative vessel stenosis and occlusion of abnormal intrathoracic vessels with coils. Other indications such as closure of atrial septal defects are about to become clinical routine, whereas interventional closure of ventricular septal defects must still be considered an experimental approach. Moreover in complex congenital heart disease there are a number of other indications for very specific interventional techniques. Interventional therapies should be seen either as adjuvant to surgical management in some cases or as an alternative approach to avoid surgery in others. It is important that the indications for surgical or interventional therapy result from a discussion involving the pediatric cardiologist and the pediatric cardiac surgeon. PMID- 9221477 TI - [Current techniques in heart surgery]. AB - Significant advances in open heart surgery during the last two decades were achieved in the field of the extracorporal circulation and the preservation of the myocardium. In the last few years, new therapeutical tools were introduced to treat patients with coronary artery disease. The transmyocardial laser revascularization (TMR) technique was introduced for clinical investigation 1990. Despite limited experience with this device in selected patients, some conclusions after a short follow-up period are available. Patients treated with TMR have significantly less anginal pain and need fewer hospitalisations. With PET follow-up studies, a better subendocardial perfusion at the expense of the subepicardial perfusion was demonstrated. On the other hand, there was no substantial increase found in terms of ejection fraction in treated patients. Minimally invasive procedures have also gained acceptance during the last few years, especially the minimally invasive coronary artery bypass procedures (MIDCAB). Introduced initially to treat solitary stenoses of the LAD without cardiopulmonary bypass, this procedure is actually often used in conjunction with PTCA for three-vessel disease in selected patients. Due to the different methods used and summarized under the term of MIDCAB, definite conclusions about the advantages of this method are difficult to formulate. There is a trend to reduced patency rates of the IMA bypasses in MIDCAB procedures compared to the conventional technique due to the difficulty with limited access. PMID- 9221478 TI - [Surgery in congenital heart defects: current developments and a few case examples]. AB - Congenital heart surgery includes the palliative treatment and surgical complete repair of cardiac malformations in newborns, children and adolescents. Palliative surgery allows early or long-term survival, depending on the primary malformation and the condition of the patient. Confection of a systemic-to-pulmonary shunt (the modified Blalock-Taussig shunt) allows in general recovery from severe cyanosis and leads to development of the hypoplastic pulmonary vascular tree in newborns with severe pulmonary stenosis. Longterm palliation can be applied to all patients in whom establishment of a biventricular heart can not be realized. The total cavo-pulmonary connection represents one possibility to bypass a single ventricle in these complex cases. Total repair allows the restitution of a completely normal anatomy and physiology after surgery; usually, this type of surgery is followed by a normalization of life expectancy with minimal pharmacotherapy. Preoperative diagnosis of congenital heart disease is reasonably performed by transthoracic echocardiography in the majority of cases. Cardiac catheterism is reserved for complex cases and those in which full hemodynamic evaluation is required for proper planning of surgery. Continuous improvement has been realized in the fields of cardiac anesthesiology and pediatric intensive care during the last decade; hence more and more complex cases have been accepted without any negative effect on the operative mortality. Additionally there has been a number of improvements in surgical and perfusion techniques, thus allowing open heart surgery in newborns with a minimal weight of 2000 g. Intraoperative transesophageal echocardiography is performed routinely and allows the proper control of surgical repair; furthermore this examination may be helpful during the weaning period from the extracorporeal circulation. There is a number of interesting topics that will take importance or will be developed in the near future: the role of interventional cardiology in pediatric patients gains more and more importance. There is a potential for minimally-invasive surgery and the number of potential candidates for heart transplantation may increase, due to the fate of long-term survivors after palliative surgery. PMID- 9221479 TI - [Club fingers and watch glass nails]. PMID- 9221480 TI - [Pulmonary mycobacteriosis]. PMID- 9221481 TI - Biochemical markers for bone diseases: current status and future trends. Proceedings of the 5th Bergmeyer Conference. Tutzing, DE, 1996. PMID- 9221482 TI - [Iodine supply at various periods in life and ultrasonographic thyroid volume in school children in a region of Switzerland]. AB - In Switzerland the previous severe iodine deficiency has been corrected by iodization of salt. Changing food habits make it mandatory to periodically monitor iodine intake. In 1994 we therefore measured thyroid volume by sonography in 217 schoolchildren, and iodine, creatinine and sodium in casual urine samples of 214 schoolchildren, 40 pregnant women and 30 breast-fed newborns. Iodine was also measured in 30 breast milk samples. In schoolchildren, goiters of WHO grades Ia/Ib/II were found by palpation in 10.6/2.3/ 0.9%. By contrast, only two of the children had a sonographic thyroid volume exceeding the 97 percentile of the WHO, which underlines the difficulty of estimating the size of small goiters by palpation. Mean thyroid volume at 6/7/9/ 10/11 years of age was 2.1/2.2/3.3/3.3/3.1 ml, which is well within the normal range suggested by the WHO. Mean urinary iodine in schoolchildren (118 +/- 49 micrograms per gram creatinine) and in pregnant women (193 +/- 113 micrograms per gram creatinine) is within required limits, albeit with a decrease since 1988. In breast milk (7.8 +/ 5.9 micrograms/dl) and in urine of newborns (6.6 +/- 3.3 micrograms/dl) iodine values are below the Swedish reference values but well above values found in iodine deficient areas. We conclude that the iodine supply in schoolchildren and pregnant women is just sufficient and in newborns slightly below recently recommended limits. Compared to 1988 the iodine supply to schoolchildren is diminishing. The discrepancy between the rather low iodine content in breast milk despite a normal iodine supply during pregnancy remains to be explained. PMID- 9221484 TI - [Cyst of the ligamentum flavum of the lumbar spine: description of 6 cases]. AB - Cysts of the ligamentum flavum of the lumbar spine have seldom been described. They are clearly visible in computed tomography as well as nuclear magnetic resonance, but are frequently wrongly diagnosed as ganglion or synovial cysts. The correct diagnosis is not feasible until after surgery. Such space occupying lesions can most often lead to uniradicular pain due to compression of a root. These cysts should be viewed as part of the degenerative process of the spine but not as tumor lesions. They need to be removed only in case of root entrapment. On the basis of six of our cases treated by surgery we describe the symptoms, imaging findings, operative techniques and pathological investigations. PMID- 9221483 TI - [Comparative effect of lansoprazole/amoxicillin with omeprazole/amoxicillin for the eradication of Helicobacter pylori in patients with duodenal ulcer]. AB - Lansoprazole, a potent antisecretory drug, possesses on an equimolar basis a 4 fold higher in vitro anti-Helicobacter pylori activity than omeprazole. In a prospective randomized study we compared lansoprazole 30 mg b.i.d. and amoxicillin 1 g b.i.d. with omeprazole 40 mg b.i.d. and amoxicillin 1 g b.i.d. for 14 days followed by lansoprazole 30 mg q.d. or omeprazole 20 mg q.d. for 14 additional days in 50 H. pylori positive duodenal ulcer patients (14f, 36m, age 27-83 [mean 43] years). H. pylori infection was diagnosed by histology (3 antral biopsies and 2 from gastric body, H & E- and Giemsa stain), rapid urease test (CLO) and culture in 39 patients, or by histology and rapid urease test in 11 patients. Control endoscopy was performed 4-6 weeks after the end of treatment. For eradication, a negative result in all 3 diagnostic modalities was required. The eradication rate was 43% (9/21 patients) in both treatment groups. 8 patients were lost to follow-up. The ulcer healing rate was 100% in both groups. Nonsmokers had a significantly higher (p = 0.026) eradication rate than smokers. No relevant adverse effects of the therapy occurred. 24 patients with persistent H. pylori infection were subsequently treated with lansoprazole 60 mg b.i.d. and amoxicillin 1 g b.i.d. for 14 days. Eradication was achieved in 5/22 (23%) patients (3/14 smokers, 2/8 nonsmokers), while 2 patients were lost to follow-up. 17 patients with persistent H. pylori infection after the second treatment received quadruple therapy consisting of metronidazole 500 mg t.i.d., tetracycline 500 mg q.i.d. bismuth-subcitrate 120 mg q.i.d. and lansoprazole 30 mg for 10 days. H. pylori eradication was achieved in 12/15 patients (80%). In conclusion, lansoprazole plus amoxicillin was equal to omeprazole plus amoxicillin in the treatment of H. pylori infected duodenal ulcer patients. Patients with eradication failure after dual therapy were successfully treated by quadruple therapy. In contrast, high dose lansoprazole and amoxicillin therapy was effective in only 23% of patients with persistent infection after standard dual therapy. PMID- 9221485 TI - [Uremia and variable swelling of the thigh]. PMID- 9221486 TI - [Calcitonin, a proximal-tubular-acting diuretic: lithium clearance measurements in humans]. AB - Lithium clearance was used to investigate the effect of calcitonin on renal sodium excretion. Two sequential renal function tests, calcitonin and placebo treated, were performed in 6 healthy students. Intravenous administration of 0.5 mg human calcitonin (hCT) (Cibacalcin, Ciba-Geigy, Basel, Switzerland) significantly raised fractional excretion of sodium (FENa), from 1.73% (0.97%) to 3.63% (0.89%) (p < 0.001), and lithium clearance (Cli), from 35.1 (5.4) to 53.9 (8.5) ml/ min x 1.73 m2 (p < 0.01). By using inulin and lithium clearance we calculated proximal (PFR), distal (DFR) and total excreted sodium (TAN). Our results clearly demonstrate that hCT has a proximal diuretic effect in humans and that a calcitonin induced proximal sodium loss is compensated by increased sodium reabsorption within 80 minutes after hCT application. Calcitonin increases the excretion of lithium and has a potential to be used in lithium intoxication. PMID- 9221487 TI - [Screening for problem alcohol drinking in the Swiss population: comparison between an ISPA-developed instrument and the CAGE questionnaire. The Swiss Institute for the Prevention of Alcoholism]. AB - In 1987, the Swiss Institute for the Prevention of Alcohol and Drug Problems (SIPA) developed a set of questions on alcohol-related problems in the general population. The aim of this article is to study the results of the questionnaire used as a screening instrument to detect problem drinking in the Swiss population, and to compare it with the CAGE test. The sample consisted of 953 people aged 20 or over. Among the drinkers (89% of the sample), 91 males (21.7%) and 34 females (8.7%) had a positive SIPA test and 53 males (12.7%) and 17 females (4.3%) a positive CAGE test. The SIPA test was more sensitive than the CAGE in detecting persons who drink regularly and quite heavily but without binge drinking. The item "Eye-opener" of the CAGE is too blunt for application to the Swiss general population and could with advantage be removed from the questionnaire. Females tend to deny alcohol problems much more than males. Binge drinking increases the risk of a positive test (SIPA: OR: 1.9; i.c. 95%: 1.2-3.0; CAGE: OR: 3.3; i.c. 95%: 1.8-6.0). In short, the SIPA test is more suitable in estimating the number of problem drinkers in the Swiss population than the CAGE, which was initially developed for the American medical population. Furthermore, the results suggest the necessity of using a different cut-off for each gender. PMID- 9221488 TI - [Fatal central pulmonary embolism under heparin therapy: white-clot syndrome]. AB - A 75-year old female underwent coronary angiography for chest pain. Significant proximal stenosis of the left coronary artery was found. During the waiting time for bypass surgery, intravenous heparin treatment was established for several days because of recurrent unstable angina pectoris. 10 days after coronary angiography an acute event with chest pain, hypotension, tachycardia and a new right bundle branch block suspect for myocardial infarction occurred, which was treated with rt-PA. Fever, persistent hypotension, acute progressive renal failure and thrombocytopenia suggested septic shock, and the patient was transferred to our hospital. A pulmonary artery catheter could not be advanced beyond the main stem of the pulmonary artery. The patient died suddenly 24 hours later from acute right ventricular failure. Autopsy demonstrated multiple white clots in both pulmonary arteries. The histological finding of clots rich in leukocytes and fibrin was compatible with the diagnosis of heparin-induced thrombosis-thrombocytopenia or white clot syndrome. Heparin-induced thrombocytopenia may occur after about 5 days of treatment. Two distinct types have been described. The first type occurs in up to 25% of patients receiving heparin and is a result of temporary platelet aggregation, margination and peripheral sequestration. The less common second type of thrombocytopenia is thought to be mediated by a heparin-dependent IgG antibody inducing platelet aggregation and may be associated with thromboembolic events leading to the white clot syndrome, which is rarely reported in the literature. In these cases heparin should be stopped immediately and replaced by oral anticoagulation. Other therapies such as low molecular weight heparin, synthetic heparinoids, hirudin, fibrinolytic agents, plasmapheresis and intravenous immunoglobulins are discussed. Monitoring of the platelet count every 5 days in patients receiving heparin for any extended period should become standard medical practice to avoid potential fatal complications. PMID- 9221489 TI - Moroto morass. A fossil ape unexpectedly resembles modern apes and humans. PMID- 9221490 TI - Curtailing the AIDS pandemic. PMID- 9221491 TI - ETS1-DNA binding and intercalation: correction. PMID- 9221493 TI - Alzheimer's maverick moves to industry. PMID- 9221492 TI - Corn genome pops out of the pack. PMID- 9221494 TI - Varmus grilled over breach of embryo research ban. PMID- 9221495 TI - Labs form biomedical network. PMID- 9221496 TI - Marijuana: harder than thought? PMID- 9221497 TI - Are pushy axons a key to spinal cord repair? PMID- 9221498 TI - Longer tusks are healthy signs. PMID- 9221499 TI - Gene discovery offers tentative clues to Parkinson's. PMID- 9221500 TI - New imaging methods provide a better view into the brain. PMID- 9221501 TI - Candid cameras for the nanoworld. PMID- 9221502 TI - Fast-action flicks draw chemists' rave reviews. PMID- 9221503 TI - Biologists get up close and personal with live cells. PMID- 9221504 TI - Spectral technique paints cells in vivid new colors. PMID- 9221505 TI - Play of light opens a new window into the body. PMID- 9221506 TI - Firefly gene lights up lab animals from inside out. PMID- 9221507 TI - Molecular individualism. PMID- 9221508 TI - Selection for survival? PMID- 9221509 TI - Learning mechanisms: the case for CaM-KII. PMID- 9221510 TI - Tissue optics. PMID- 9221512 TI - [Age determination by measurement of the ratio of D- and L- forms of aspartic acid. II. Chiral separation of amino acids from human dentin]. AB - Estimation of relation of D,L-forms aspartic acid has been recently used for an evaluation of age. It represents a relatively new method based on analysis of hydrolyzed dentine. Its amino acids are derived and analyzed by gas chromatography or liquid chromatography on a chiral column. PMID- 9221511 TI - Proceedings of the roundtable of experts in surgery blood management. Vienna, April 7-9, 1995. PMID- 9221513 TI - [Polychlorinated biphenyls in human subepicardial fat]. AB - A considerable relation between myocardial fatty infiltration (lipomatosis) and ischemic lesion of myocardium was proved by authors else where. This time, polychlorinated biphenyls (PCB) wee studied in subepicardial fat as well as in subcutaneous fat in thorax and abdomen of deceased person after sudden and violent death. For the time being, most published results concerned the abdominal subcutaneous fat. All the published results showed high concentrations of PCB were proved in subcutaneous fat tissue from thorax and abdominal area and specially in subepicardial fat. Topical relation of PCB deposits and heart muscle was direct in all analyzed case because of fatty infiltration of myocardium. Obviously, a direct toxic effect of PCB on myocardium can take place in stress lypolysis e.g. associated with heart infarct (in some animals hydropericardium and impairment of parenchymatous organs were observed) and prognosis of patients with ischemic lesion can get worser. PMID- 9221514 TI - [Detection of 11-nor-delta-9-tetrahydrocannabinol-9-carboxylic acid in urine using thin-layer chromatography]. AB - 11-nor-delta-9-tetrahydrocannabinol-9-carboxylic acid is the biotransformation product of delta-9-tetrahydrocannabinol, that is the main active compound of the product from cannabis: marihuana and hashish. Its identification in urine is supposed to be the best indication of previous cannabis consumption. The aim of this study was to find conditions for the use of thin layer chromatography as an identification method of 11-nor-delta-9-tetrahydrocannabinol-9-carboxylic in urine after positive preliminary screening of cannabinoids by EMIT method. PMID- 9221515 TI - [Present possibilities of age determination in forensic medicine with emphasis on the importance of measurement of D- and L- forms of aspartic acid. I. An overview]. AB - Evaluation of age of unknown deceased persons belongs to the most important ways to identification. For the time being, morphological methods are used, namely evaluation of age according to Gustafson's method from tooth grindings or by macroscopical estimation of abrasion, transparency of root dentine, alveolar atrophy and number of missing teeth. Evaluation of the data can be influenced by an individual failure and experience showed an age related decrease of precision. Recently, some papers occurred estimating a relation of D, L-forms of aspartic acid which depends on the age with a significant precision. PMID- 9221516 TI - [An overview of the symptoms and signs of voice disorders and the pathophysiology of hoarseness]. AB - Voice disorders are due to organic and functional disturbances of the voice generator, activator and resonator. They appear as a consequence of different factors which lead to the development of hoarseness, and may as well be the result of disturbed phonatory patterns. Phonatory patterns refer primarily to the muscular activity of the vocal system, which is delicately balanced within the voice generator, activator and resonator. PATHOPHYSIOLOGY OF HOARSENESS: Basic causes of hoarseness are insufficient glottic closure during phonation (glottal gap), changes in the vocal fold stiffness and imbalance in mechanical properties between the two folds. Glottal gap leads to the excessive air leakage during phonalion and insufficient conversion of the expiratory air into pulses. Turbulence of the expiratory air particles is increased, leading to the development of noise. Excessively stiff or tax vocal folds, both disturb the vibration process and lead to the development of noise and hoarseness. Imbalance in tension between the two folds, and especially in their mass, may lead to the glottal gap, with the consecutive noise and hoarseness. PHONATORY PATTERNS: Phonatory patterns refer to habitual movements of the vocal organs during phonation and speech, which are acquired during the process of learning phonation and speech. This is primarily the muscular activity of generator, activator and resonator of the voice, which is so balanced to produce the optimal voice quality with the least effort and fatigue. The activity of the phonatory organs is not well balanced in cases of voice disorders. That is the primary cause of functional voice disorders, and a very frequent consequence of organic voice disturbances. Hyperkinetic dysphonia is the most common type of disturbed phonatory patterns, characterized by excessive vocal effort, while hypokinetic dysphonia is rarely seen. The third type of functional disorders of the phonatory patterns is an incorrect placement of the voice (imposlatio falsa), which is characterized by an imbalanced muscular activity of the vocal organs, but within the normal limits concerning the overall amount of activity. PMID- 9221517 TI - [On the threshold of new treatment of diabetic retinopathy]. AB - The introduction of ocular photocoagulation, almost thirty years ago, was the first successful prevention of blindness from diabetic retinopathy in some patients. The development of lasers, and the growing knowledge of indications for their application, reached the point at which the growth of new blood vessels could be stopped or reversed, and legal blindness from macular oedema avoided in about half of the treated eyes. The attempts at the very prevention of diabetic retinal microvascular complications at some more physiological, even molecular level, aside from the palliative treatment by photocogulation, brought some new and exciting results which are reviewed here. First, and foremost, is the understanding of the importance of the tight blood glucose control which, if started early and kept long enough, slows down the development of retinal lesions and offers a better prognosis for vision in a substantial number of diabetic patients. Second, the development of a unique animal model of proliferative retinopathy which mirrors human disease both chemically and histopathologically offers a field for investigation of both pathogenesis and therapy of this most dreadful complication of diabetes. Finally, there is a bulk of new evidences about the key role of vascular endothelial growth factor/vascular permeability factor in ocular angiogenesis which will probably result in the new approach to the prevention of neovascular growth by inhibition or modulation of VEGF/VPF activity. PMID- 9221518 TI - [Leiomyoma of the esophagus. Case report]. AB - Of all oesophageal tumours benign tumours account for less than 10%, of which 4% are leiomyomas. These tumours are most frequently asymptomatic, mostly localized in the lower oesophageal third. The most frequent symptoms, if any, are the following: dysphagia, unspecific retrostemal pain, heartburn, and occasionally, weight loss. Tumour enucleation is a therapy of choice in patients with oesophageal leiomyoma. In case of successful surgical removal, the prognosis is good and complains are practically eliminated. A male patient, aged 53, with paroxysmal tachycardia, in whom transhiatal enucleation was carried out in order to remove a large oesophageal leiomyoma after which cardiac complains were eliminated, is reported. PMID- 9221519 TI - [Interactions of the most frequently used drugs in the treatment of angina pectoris]. AB - It seems that knowledge and evaluation of drug-interactions with organic nitrates, beta-blockers and calcium-channel blockers are of great importance in relation to patients with chronic diseases, who usually take more than one drug, which is in conjunction with the effect of their ageing. Organic nitrates are safe and the most important drug-group in angina pectoris therapy. Beta adrenoreceptor blocking agents and calcium-channel blockers are involved in interactions with many different drugs (twenty interactions), but only three interactions are evaluated as interactions of high clinical significance (adrenaline, cimetidine and anti-thyroid agents with beta-blockers, and anti arrhythmic agents, beta-blockers and cyclosporin with calcium-channel blockers), while the others are of moderate or minimal significance. The most desired interactions of these drugs can be avoided: adjustment of dosage, avoidance of fixed-combination, use of adequate form of drug, or alternative drug and, if necessary, elimination of one drug from therapy. Knowledge of these drug interactions is important for physician's routine practice either in primary health-care or in hospital conditions. PMID- 9221520 TI - [Metastasis in axillary lymph nodes in breast carcinoma--possibilities of mammographic diagnosis]. AB - INTRODUCTION: Enlarged axillary lymph nodes are often found during routine examinations. They are usually circular, oval, lobular, smaller than 1 cm, with typically changed fat centers. The condition of axillary lymph nodes in breast cancer is of great importance for timely diagnosis, because its metastatic spread is a primary prognostic sign of breast cancer. Therefore, the purpose of this paper is to explore possibilities of mammographic diagnosis in detecting enlarged lymph nodes. MATERIAL AND METHODS: The authors present 69 patients with enlarged axillary lymph nodes discovered by clinical examination and mammography of the axila. Of 69 patients, 47 patients had metastatic breast cancer, 21 subject had dysplasia and 1 patient had bacterial infection. Of 47 patients with breast cancer metastases, in 38 the tumour mass in breast was visible by mammography; microcalcifications had been found in 5 patients, while in 4 subjects the tumour mass was not visible either in one, or the other mamma. Biopsy was performed in all 47 patients and diagnosis was confirmed microscopically (by PH). DISCUSSION AND CONCLUSION: The appearance of enlarged lymph nodes in axilla may be a primary clinical and mammographic sign of the earliest breast cancer. The diagnosis is not the problem if lymphadenopathy, together with visible cancer, is present (Figure 1). The problem arises when enlarged lymph nodes are without visible tumour mass in the breast (Figures 2 and 3), because their enlargement can also be seen in dysplasia, inflammation (Fig. 6), lymphoma, metastases of some other tumour (rarely), systemic disease. In all presented patients with metastatic breast cancer, lymph nodes were extremely enlarged, homogeneous and separated. All of them had no hylar fatty degeneration which is a characteristic of benign enlargement of lymph nodes. But, it is impossible to make a differential diagnosis between malignant changes and benign enlargement of lymph nodes by mammography. Consequently, biopsy of all lymph nodes larger than 1 cm, being not infiltrated by fat is suggested (excluding mastitis and dermatitis), because only pathology can give answer to the cause of lymph nodes enlargement. PMID- 9221521 TI - [Thrombophilia, prethrombotic conditions, hypercoagulability]. AB - The editorial deals with the state of art from the terminological point of view a very important problem of thrombosis and thromboembolic states. According to world medical statistics these diseases are one of the most frequent causes of illness and death. The rationale for the writing of this editorial is imposed by many controversies in Serbian medical literature related to diagnosis, treatment and especially prophylaxis of these pathologic states and the state of the art of the above mention problems in foreign medical literature. On the basis of the study of the state of the art in foreign medical literature. It could be concluded that the biochemical basis of thrombophilia, pre-thrombotic states and hypercoagulability is, in fact, the same phenomenon with regard to the disturbed dynamic balance between activating factors of haemostatic system and inhibitory factors. Activating factors of haemostatic system are prevailing in these pathological condition. The difference is express, however, in a different clinical feature of this unbalance which lead, earlier or later, to thrombosis. It is emphasized that markers which predicts occurrence of thrombosis have not yet been recognized and that the significance of the activation of the haemostatic system have to be established by multicentre investigations, i.e. that the role of these markers in the development of thrombotic diseases should be established. At the same time, on the review of the current Serbian medical literature, it is concluded due to lack of technical prerequisites the current doctrinaire definition of the above mentioned states can not be carried out in our medical institutions and laboratories. PMID- 9221523 TI - [Delayed-onset dystonia due to asphyxia in the perinatal period]. AB - The phenomenon of delayed-onset dystonia following presumed "static" brain injuries was described after stroke and head trauma. Burke et al. described a different category of secondary dystonia, where perinatal injury (asphyxia) caused minimal or no immediate neurological deficit, with the delay of years before dystonia emerged. This type of dystonia following perinatal injury has been termed "delayed onset dystonia due to static encephalopathy of childhood". According to the definition of dystonia, we were able to select 5 patients with the aetiologic diagnosis of perinatal asphyxia from the group of 347 out- and inpatients (1.4%) treated for various types of dystonia at the Movement Disorders Department (Institute of Neurology, CCS, Belgrade) from November 1986 to November 1994. At onset of dystonia the mean age of patients was 13.2 years (range from 10 to 17), with combined initial involvement of the arm and neck in 3 patients. The period from the onset of the disease to the maximum severity lasted 8.2 years (range from 4 to 14), resulting in segmental brachial dystonia in 3, hemidystonia and generalized dystonia in one patient each (Table 1). The adverse perinatal events are described in Table 2. Three of our patients had delayed achievements of developmental milestones. All patients were regularly schooled and had preserved intellectual capacities, except the patient 3 whose achievements were below average (IQ = 86). Different drugs were administered (Table 3), but moderate effects were achieved only with trihexyphenidyl in two patients (daily doses of 24 mg and 30 mg, respectively), and baclofen (80 mg p.d.) in one patient. In this study we describe 5 new patients who fulfilled the criteria for the diagnosis of delayed-onset dystonia due to perinatal asphyxia (Tables 1 and 2). We accepted the approach of Saint-Hilaire et al. to suggest a relationship between perinatal asphyxia and later occurrence of dystonia in our 5 patients. However, coincident occurrence of a primary dystonia with a static encephalopathy of childhood due to perinatal asphyxia cannot be excluded. This phenomenon of delayed appearance of dystonia was also described in other forms of static cerebral injury; i.e. stroke, head trauma or anoxic brain damage. Interestingly enough, age at the time of anoxia or brain insults seemed to be crucial for the development of dystonia: those who suffer acute brain insults during childhood or early life are more likely to develop dystonia than the older patients. Therefore, the "static" nature of encephalopathy induced by perinatal asphyxia is questionable. Finally, this study strengthens the suggestion that perinatal asphyxia can lead to delayed-onset dystonia, and, since "some of these patients closely resemble cases of idiopathic torsion dystonia, the prior occurrence of asphyxia should be used as a criterion of exclusion for that diagnosis". PMID- 9221522 TI - [Dacron and polytetrafluoroethylene aorto-bifemoral grafts]. AB - INTRODUCTION: In reconstructive procedures of the abdominal aorta synthetic grafts are today mostly used. There are two types of bifurcated synthetic grafts: Dacron and polytetrafluorethilene (PTFE). In many papers these grafts are compared in aortobifemoral position. Karner 1988, and Lord 1988, found no significant difference between them after aortobifemoral reconstructions. In 1955. Paaske wrote about a new "stretch" bifurcated PTFE graft in aortobifemoral position. Comparing this material with standard Dacron graft, he only found a shorter operating time. The aim of this paper is to compare Dacron and PTFE bifurcated grafts in aortobifemoral position in patients with aortoiliac occlusive diseases. MATERIAL AND METHODS: This prospective study included 283 aortobifemoral reconstructions due to aortoiliac occlusive diseases operated between January 1st, 1984 and December 31st, 1992 at the Institute for Cardiovascular Diseases of the Serbian Clinical Centre in Belgrade. Bifurcated PTFE grafts were used in 136 patients, and nonimpregnated knitted Dacron grafts in 147 subjects. There were 25 (8.8%) female and 258 (91.2%) male patients, average age 56.88 years. Ninety one (32.2%) patients had a claudication discomfort (Fonten stadium II), 91 (32.2%) disabling claudication discomfort (Fonten stadium IIB), 45 (15.9%) rest pain (Fonten stadium III), and 56 (19.8%) gangrene (Fonten stadium IV). In 45 (15.9%) patients previous vascular procedures were performed. Prior to operation, Doppler ultrasonography and translumbar aortography were carried out (Figure 1). Transperitoneal approach to abdominal aorta, and standard inguinal approach to femoral arteries were used. In 154 (54.4%) patients proximal anastomosis had an end to side (TL), and in 129 (45.6%) end to end (TT) form. In 152 (26.88%) cases distal anastomosis was done in the common femoral (AFC) artery, and in 414 (73.2%) cases in the deep femoral (APF) artery. In 7 patients the aorto-femoro-popliteal "jumping" bypass was done, and in 29 patients simultaneous sequential femoro-popliteal bypass graft. The patients were following-up over the period from one, six and twelve months after operation, and later once a year, using physical examination and Doppler ultrasonography. In patients with suspected graft occlusion, anastomotic stenosis, pseudoaneurysms, progression of distal arterial diseases, Duplex ultrasonography and angiography were also used, and leukoscintigraphy in patients with suspected infection. Statistical analysis was performed using Long Rank and Student t-test. RESULTS: Inhospital mortality rate was 11 (7%). Distal reconstructions significantly increased the mortality rate when simultaneously performed with aortobifemoral bypass graft (p < 0.01). The follow-up period was from 2 months to 9.5 years (mean 3.6 years). The early patency rate was 97% from PTFE and 99.4% for Dacron grafts, while the late patency rate was 94.9% for PTFE and 96.6% for Dacron grafts. The type of the graft had no statistical influence on the early and late graft patency (p > 0.05) (Graphs 1, 2, 3). Six (2.1%) early unilateral limb occlusions were observed. Five patients had the PTFE and one the Dacron graft, without statistically significant difference (p > 0.05). The reasons for early graft occlusion were: stenosis of distal anastomosis in 3 patients, and pure run off in 3 patients. In 5 patients urgent reoperation (limb thrombectomy with profundoplasty or femoro-popliteal bypass graft above the knee) were done with complete recovery of legs. However, in one patient the above knee amputation was done. During the follow-up period, 14 (5.2%) late graft occlusions were recorded. There were 11 unilateral limb occlusions and 3 bilateral. All patients with bilateral occlusions had PTFE grafts but this was not statistically significant (p > 0.05) comparing two types of grafts. Taking into account all late occlusions, there were 7 PTFE and 7 Dacron grafts. There was no statistical difference betwe PMID- 9221524 TI - [Phoniatric surgery and conservative treatment of vocal cord hematoma]. AB - INTRODUCTION: Functional-traumatic lesions of the vocal fold include mucous stranding, "nodular" lesions, polyps, cysts, contact hyperplasia and haematoma of the vocal fold. An acute voice overuse may result in bleeding (haematoma) within the vocal fold. This may be in the form of petechial bleeding, or a genuine haematoma develops within the tissues of the vocal fold. Haematoma may also arise as a consequence of prolonged cough, forceful vomiting, lifting of a heavy weight, various effortful activities, etc. Haematoma is usually located close to the vocal fold free edge and therefore disturbs the glottic closure during phonation. The treatment is adapted to the size and localization of haematoma, as well as to the time elapsed from onset of the lesion. Phonosurgery can be used in therapy, as well as corticosteroid treatment. MATERIAL AND METHOD: A series of 102 vocal fold haematomas has been treated by phonosurgery (39) and conservative therapy (63). Phonosurgical interventions were performed by an indirect approach, by use of microstroboscopy (28 patients) and videostroboscopy (11 causes). Conservative treatment consisted of corticosteroid therapy. RESULTS: During a 10 year period 1550 phonosurgical operations were performed for benign lesions of the vocal fold, including 39 haematomas (2.5%). It was established that recovery of vibration pattern was significantly faster in the surgery group in comparison to the group of patients treated conservatively. All surgical patients were operated within the first several days after the onset of symptoms. DISCUSSION: In case of a vocal fold haematoma, it is very important to establish the diagnosis as soon as possible in order to start with the therapy early enough. Within the first several days after the onset (the best within 24-48 hours) a phonosurgical treatment is indicated, preferably by the use of indirect videostroboscopy. If the treatment is started later we use corticoids. However, the results are inferior as compared to surgery. We did not perform direct microlaryngoscopy in these cases, for a lack of function monitoring and possible local trauma to the tissues. In the majority of cases the voice therapy is required as well. PMID- 9221526 TI - [New trends in immunosuppressive therapy in patients with kidney transplants]. AB - Following the transplantation of visceral organs, two populations of immunocytes survive which derived from the donor's and the recipient's bone marrow. The rejection process of transplanted organ or graft versus host reaction could result from their mutual stimulation. On the other hand, the immunocytes of the donor and recipient could deny the effects of their MHC antigen disparities directed against each other, to favor the transplant acceptance through the processes of the donor specific tolerance. The spontaneous chimerism has been shown even in patients with the heart or kidney allografts which contain few numbers of donor's passenger lymphocytes. The frequency of chimerism is augmenting more than 1000 times following the combined and simultaneous transplantation of both kidney allograft and donor bone marrow cells. Theoretically, the continuous peroraly giving of high doses of synthetic peptides homologous to immunodominate epitope(s) of the donor MHC molecules could sustain the state of "stable chimerism" with donor immunocytes. This immunosuppressive protocol permits long-term survival of organ (kidney) allograft without diminishing the donor immune system that provokes the present immunosuppressives with non-specific effects. PMID- 9221525 TI - [The effect of extent of tumor resection on the outcome of combined therapy in patients with glioblastoma multiforme]. AB - INTRODUCTION: The importance of the extent of surgery as a prognostic factor in multiform glioblastoma has been investigated for years. Some studies could not establish its influence on survival of patients treated with surgery, postoperative radiotherapy, with or without chemotherapy. On the other hand, there are data suggesting benefit for patients treated with more aggressive surgical approach. The aim of this study was to investigate the influence of the extent of surgery on survival/progression-free survival of patients with multiform glioblastoma treated with two consecutive protocols of a combined approach. MATERIAL AND METHODS: Of 86 patients that entered this study, thirty seven were treated with surgery, postoperative hyperfractionated radiotherapy using 1.2 Gy b.i.d. to a total tumour dose of 72 Gy in 60 fractions in 30 treatment days and adjuvant chemotherapy consisting of BCNU, vincristine, procarbazine and cisplatin for up to 6 cycles or until tumour progression. Forty nine patients were treated with surgery and postoperative accelerated hyperfractionated radiotherapy using 1.5 Gy b.i.d. fractions to a total tumour dose of 66 Gy in 44 fractions during 22 treatment days. BCNU and hydroxyurea were given once weekly during the irradiation period. Surgery consisted of biopsy in 25 patients and subtotal or gross total tumour resection in 61 patients. Patients treated with a more radical surgery had longer median survival time and higher 1- and 2-year survival rates than those treated with biopsy (56 v.s. 29 weeks, respectively; 62% and 23% v.s. 16% and 0%, respectively; long rank, p = 0.0000) (Figure 1). They also had longer median time to tumour progression and higher 1 year progression-free survival rate than those treated with biopsy only (33 v.s. 21 weeks, respectively; 20% v.s. 0%, respectively; log rank, p = 0.00000) (Figure 2). Multivariate analyses using both survival and progression-free survival as endpoints confirmed that the extent of surgery was an independent prognostic factor, together with the age, tumour location, and interfraction interval (Tables 3 and 4). DISCUSSION: The benefit of a more radical surgery remains controversial in patients with multiform glioblastoma, although maximal tumour reduction should be supported from the cytokinetic point of view. Findings of various authors support this view. Results of this study add further evidence that the aggressive surgical approach carries significant benefit for patients with multiform glioblastoma regarding the survival and progression-free survival. These observations are confirmed with multivariate analyses that showed independent influence of this prognostic factor. PMID- 9221527 TI - [Up-to-date study of rheumatoid factor and the clinical significance]. PMID- 9221528 TI - [Clinical medicine of 21st century and rheumatology]. PMID- 9221529 TI - [Rheumatology and pediatrics]. PMID- 9221530 TI - [Therapeutic planning of patients with early-stage arthritis rheumatoid]. PMID- 9221532 TI - [Therapy of arthritis patients with spinal lesions]. PMID- 9221531 TI - [Endocrine disorders in patients with arthritis rheumatoid]. PMID- 9221534 TI - [Is it possible to diagnosis patients with early-stage arthritis?]. PMID- 9221533 TI - [Lecture on the therapy of rheumatism for practice physicians]. PMID- 9221535 TI - [Mechanism of osteoclastic bone loss of articular cartilages in patients with arthritis]. PMID- 9221536 TI - [Orthotic devices for patients with rheumatism]. PMID- 9221537 TI - [Proctology, the science of diseases concerning the anus]. PMID- 9221538 TI - [The arduous way to the stool--the history of constipation]. PMID- 9221539 TI - [Defecation problems: incontinence, constipation and impeded defecation; why and what can be done?]. AB - Defaecation disorders may be subsumed in three categories: Inability to control motions = incontinence. Difficulty of evacuation = constipation [inertia coli, outlet obstruction]. Impeded defaecation: Rectocele, enterocele, intussusception. Etiology, examination and therapy are described in detail. Characteristic complaints of patients are listed and matched with probable diagnoses. Beside routine proctologic examination endosonography, estimation of transit time, endoscopy and defecography are discussed. The role of nutrition is stressed and emphasis layed on fibre and fluid intake. The advice, "take more fluid and fibres" does not help a lot, because no individual help is given. A time consuming nutrition and defaecation history has to be taken to establish nutritional support. This attention gives confidence to the patient and helps a great deal in the treatment. A checklist of the therapy of constipation and summarizing tables on different types of fibres are included. Additional conservative treatments are pelvic exercises and biofeedback training. Operative therapy is directed towards etiology of the disorder. Therefore many different methods exist and their diagnose related indication are discussed. PMID- 9221540 TI - [Hemorrhoids--etiology, symptoms and therapy]. AB - Hemorrhoids are a very widespread disease causing pain by thrombosis, fear by bleeding and be a burden by weeping and pruritus. The different treatments show the different interpretations of physicians and patients. This paper will give a view of the most common and standardized procedures. Rubber band ligation is the most effective procedure in treating 1st and 2nd grade hemorrhoids. Surgical procedure is best in 2nd and 3rd grade disease. Recurrency is between 0.5 and 5% only. Technical development in surgical practice was leading to less pain in postoperative period by using diathermy, non-traumatic technique and avoiding of tampons. Using these principles hemorrhoidectomy allows a short hospital stay according to the individual needs of the patient. Special care should be given on urinary retention, the most common complication in anorectal surgery. PMID- 9221541 TI - [Anal fissure]. AB - The anal fissure is a very common disease of the anal canal. Diagnosis is easy and most patients have a typical history of pains and anal bleeding. Proctoscopy will certify the anal fissure. The treatment is based on reducing the spastic of the internal anal sphincter, either by dilating the anal canal or sphincterotomy. Excision of the anal fissure should only be done in case of fissures with intersphincteric fistula or abscess. High fibre diet and increasing the volume of daily drinks is a useful and very efficient help in treating anal fissures. PMID- 9221542 TI - [Rectal prolapse]. AB - Rectal prolapse is the transposition of the entire rectal wall into the rectal lumen, the anal canal or through the anal canal out side. It differs from anal prolapse in thickness, circular plication of the mucosa and, if large, its extent. The cause is not clearly established, but disorders in bowel movement seem to be of importance. Symptoms reach from the feeling of incomplete evacuation to defecation block and irreducible prolapse. The diagnosis of outer prolapse is easy. The inner prolapse [intussusception] can be suspected by anamnesis and in the presence of solitary rectal ulcer. Defecography gives the conclusive examination. Conservative therapy is analogous to hemorrhoids: Fibres and sufficient liquid intake. Operative procedures can be divided in transabdominal and perineal procedures. From the latter Delorme's procedure gives good results with low stress for the patient. Of the transabdominal procedures we favor rectopexy with Ivalon-sponge, preservation of the lateral bands and sigmoid resection. This procedure can easily be done by laparoscopy. Postoperative constipation is observed above all if the lateral bands are dissected and no sigmoid resection is done. Preexistent constipation Improves in about 50% of the cases. Same does incontinence. PMID- 9221543 TI - [Anorectal abscess and fistula]. AB - Anal glands which lie in the intersphincteric space at the dentate line play an important role in the development of abscesses and fistulas. Secondary causes are inflammatory colorectal diseases and skin infections. For the classification of anorectal abscesses and fistulas a simple and practible nomenclature is proposed. Clinical presentation and diagnostic are described with regard to new means of diagnostic. The opening of the abscess without a fistulotomy appears to be the treatment of choice in patients with a perianal abscess whereas patients with an intersphincteric abscess are treated by fistulotomy. Perianal fistulas can be detected in 50% of the cases after drainage of a perianal abscess. The therapy of the perianal fistula below the dentate line consists in the fistulotomy. In contrast intersphincteric fistulas and fistulas above the dentate line are more often treated with a mucosa flap where by the sphincteric musculature is spared. PMID- 9221544 TI - [Anal and perianal tumors]. AB - The anal canal extends from the anorectal ring to the anal verge. Different kinds of epithelium are existing. Below the dentate line is squamous epithelium, above columnar. In between is a gradual transition area, the so called transitional zone. According to the World Health Organization the anal margin is outside the anal verge and the anal canal reaches up to the superior border of the levator muscle. The lymphatic drainage of the anal margin is to the inguinal lymph nodes, from the anal canal to the inferior mesenteric nodes. Tumors of interest in the perianal location are Paget's disease and Bowen's disease as an intraepithelial adenocarcinoma or a squamous cell carcinoma in situ. Wide local excision is the treatment of choice. The true epidermoid carcinoma can be found at the anal margin and the anal canal. The symptoms are mild and unspecific. Many tumors are diagnosed late. The standard treatment has changed in the last 20 years. Local excision and abdominoperineal resections were followed by a high rate of local recurrence. The 5 year survival rate was 50% overall. The treatment of choice today is a combined chemoradiation therapy following Nigro's recommendation. The 5 year survival rate is approximately 85%. PMID- 9221545 TI - [Perianal dermatitis]. AB - The most often seen perianal dermatitis is the eczema, which is caused symptomatically by haemorrhoids, fissures, fistulas, proctitis, contact allergies, the funnel shaped anus and intensified folding of the perianal skin. Before treatment of the eczema all these causes have to be eliminated. Differential diagnosis of the perianal dermatitis are psoriasis, seborrheic dermatitis, endogenous eczema, intertrigo, proctitis and skin cancer. PMID- 9221546 TI - [Case of the month]. PMID- 9221547 TI - [Idiopathic pericardial effusion with tamponade in a Friesian gelding]. AB - A 7-year-old Friesian gelding was referred to the Department of Large Animal Medicine and Nutrition of the Faculty of Veterinary Medicine because of an inadequate response to treatment for vague symptoms of colic. An extensive physical examination showed there to be circulatory problems, with right-sided decompensation. Heart sounds were muffled on both sides of the thorax, and ECG showed a low voltage and variable amplitude of the QRS complex. Further investigations indicated idiopathic pericardial effusion. Pericardiocentesis was performed and supportive therapy started. Three weeks later the circulatory system no longer showed abnormalities and the patient was discharged. When a patient presents with vague 'colicky' symptoms that cannot be explained by abnormalities of the digestive tract, the differential diagnosis should include the possibility of 'false' colic, of which pericardial effusion is one possible cause. Early diagnosis and treatment can contribute to a good result. PMID- 9221548 TI - [Homeopathy in veterinary medicine]. PMID- 9221549 TI - [Helpful chemistry. Professor A. Ruiter's farewell lecture on 7 November 1966]. PMID- 9221550 TI - [15th International Symposium of Salmonellosis and Brucellosis: a report]. PMID- 9221552 TI - [Wound care in companion animals]. PMID- 9221553 TI - [KNMVD's involvement in swine fever. Provision of information is essential for control. Royal Dutch Society of Veterinary Medicine]. PMID- 9221551 TI - [A frequently used drug with fatal outcome!]. PMID- 9221554 TI - [Quality of health care: at odds with suitability]. PMID- 9221555 TI - [Paradoxical welfare perception by the baby boom generation around 2025. 3 scenarios for pension and care]. AB - Public pension (AOW), supplementary pensions and care-arrangements together constitute the important financial arrangements for the elderly in the Netherlands. The ageing process jeopardizes them all, although the uncertainty about the future development of supplementary pensions is huge. The problems with the care provision will probably be bigger than those for the public pension, because other factors (like age-dependent use of care and insufficient growth of labour productivity in the care sector) add to the growth of expenses due to the rising share of elderly persons in the population. Therefore, contributions of the elderly will be necessary, collectively or individually. A growing standard of living, shifting the criteria for an acceptable minimum standard, both in income and care, sets a major problem. The consequences of changes in the standard of living and in the perception of acceptable minimal standards are explored in three scenarios. If the standard of living does not grow much (scenario 1), future elderly persons will be more prosperous compared to younger individuals and their capacity to contribute to the growing expenses for care will grow. If however the standard of living grows strongly (scenario 2 and 3), future elderly persons will impoverish compared to younger adults. Furthermore, their ability to pay will diminish. This is even more so the case, as in this situation the prices of care will be higher. If the public pension becomes the most important component in the pension system, the incomes of most retired people will be near a relatively high social minimum level. However, if supplementary pensions become most important, the differences in wealth within the retired population will be marked. PMID- 9221556 TI - [Peripheral nerve stimulation in Alzheimer's disease]. AB - The "use it or lose it' concept implies that stimulation of neurons might stop degenerative activities and initiate regenerative processes in aging and Alzheimer's disease (AD). Based on this concept, the effects of Transcutaneous Electrical Nerve Stimulation (TENS), tactile stimulation, and a combination of the two on memory and affective behaviour of AD patients were examined. The results suggest that, compared to AD patients of the control group, patients of the experimental group improved in visual short-term memory, verbal and visual long-term memory, and verbal fluency. Moreover, stimulated AD patients participated more independently in activities of daily life and their affective behaviour improved. As in those studies the therapist was present during the peripheral stimulation of the experimental group and the sham stimulation of the control group, interpersonal communication alone could not explain the treatment effects. However, a positive effect of the combination of TENS with personal interpersonal communication could not be excluded. Consequently, it was examined whether TENS, in the absence of the therapist, could also have a beneficial influence on the cognitive and behavioural functioning of AD patients. In addition, it was investigated whether TENS had a positive effect on the rest activity rhythm of AD-patients. The results show that improvements in visual short- and long-term memory, verbal long-term memory, and verbal fluency are solely due to the peripheral stimulus itself. Furthermore, the independent and affective functioning of both the experimental and control group appeared to relatively improve by interpersonal communication. Moreover, the rest-activity rhythm of stimulated AD-patients improved. Peripheral nerve stimulation in AD might thus become a new treatment strategy to improve patients' quality of life. PMID- 9221557 TI - [Depression in the elderly in family practice]. AB - Depression among the elderly is an important problem in general practice. There is insufficient knowledge of the prevalence of this condition, of possible ways to improve diagnosis and of its course. The questions in this project were: a) how prevalent is depression in the active population of the GP, and which patients are identified as such by the GP, b) how can the depressed elderly patient be diagnosed in daily practice and which signals can be used by the GP to include or exclude the diagnosis, and c) what is the recovery of this condition among the elderly and is recovery related to recognition by the GP? a) The prevalence has been estimated in two studies (n = 384, n = 580) in 1990 en 1992 with self-report questionnaires and interviews. The samples consisted of consecutive elderly attenders of general practices. We used the Zung scale, the Geriatric Depression Scale (GDS) and the Diagnostic Interview Schedule (DIS). b) To improve diagnosis the relationship of various characteristics to depression status at interview in 1992 was analyzed, and we also evaluated whether the 30 item version of the GDS could be shortened for use in daily practice. c) Patients with depression at baseline interview in 1992 were reassessed with the DIS at 6 and 12 months to evaluate the course. a) Many elderly patients indicated depressive complaints on the self-report questionnaires for depression, respectively 11% and 13% for Zung and GDS. One in twelve elderly patients had depression according to the interview in 1992 (46/580). Not all patients who have depressive complaints or depression were recognised by the GPs. b) Vague or gastrointestinal (GI) reasons for visiting the GP or presenting symptoms and female gender were related to depression. To make suspected depression more likely or to exclude depression, 4 questions may suffice. c) At 12 months 72% of the 25 patients with depression at baseline interview were not depressed. Whether patients were depressed at follow-up interviews was not related to recognition by the GP. The conclusion is that vague complaints and GI problems may be a signal for depression in the elderly and a few short questions may make this diagnosis more likely or exclude it. The course of this condition in general practice needs further study. PMID- 9221558 TI - [Gait impairment in the oldest old]. AB - To evaluate senile gait patterns in octogenarians and nonagenarians, we provided a standardized questionnaire on gait disabilities to 153 elderly subjects over 88 years of age. Subjects represented a relatively healthy subgroup of non institutionalized residents who participated in a gerontological survey of all inhabitants of the city of Leiden who were 85 years or older. Of the 142 subjects who responded to this questionnaire, 87 persons (61%) claimed distinct diseases as a cause of gait impairment. Of the remaining 55 persons, 42 received a standardized gait assessment. Gait was classified as completely normal in 25 persons (18% of all responders), whereas in three other persons gait could not reliably be classified as either normal or abnormal. A wide spectrum of clear gait abnormalities-mainly with ataxic features-was encountered in the remaining 14 persons (10%). It is concluded that some elderly subjects have a mainly ataxic gait disturbance which seems unrelated to the presence of distinct diseases. Although additional investigations might still reveal underlying pathology in these subjects, their gait impairment may represent the "idiopathic senile gait disorder'. In addition, a relatively high number of very old community residents have a completely normal gait. PMID- 9221560 TI - [The effect of chemotherapy on energy and nitrogen balance in patients with hematologic neoplasms]. AB - Energy balance and nitrogen balance were evaluated within the opening week of standard induction chemotherapy in 26 haematooncological patients. The patients were uncomplicated in good nutritional status and nutritional requirements were covered by oral diet under the daily assistance of specially trained dietary nurse. Resting energy expenditure (REE) measured by indirect calorimetry under standard circumstances was elevated to 113.1% of predicted value by Harris Benedict equation. We found a significant decrease in REE to 106.1% of predicted value (p < 0.01) on day 7 after the beginning of induction chemotherapy. Total energy requirements calculated on the basis of measured REE were not elevated during chemotherapy and mean energy balance was balanced. On the other hand mean nitrogen balance was markedly negative during chemotherapy even on the second day of treatment (-6.9 gN/day, cumulative nitrogen balance -28.0 gN/5 days). The negativity correlated will with markedly elevated urinary nitrogen output but worse with nitrogen intake in the diet. Significant correlation was found between the negativity of cumulative nitrogen balance for the whole period of follow up and the magnitude of decrease in REE after chemotherapy (r = 0.74, p < 0.01). This dependence may give evidence of the decay of tumor mass as the main factor of changes shown. Findings described here may support the assumption that energy requirements of haematooncological patients in good nutrition status during chemotherapy may be covered by oral diet even though this usually does not prevent the negativity of nitrogen balance. PMID- 9221559 TI - [Interferon-alpha in the treatment of patients with chronic myeloid leukemia]. AB - A retrospective analysis of the treatment with Interferon alpha in 18 patients with Ph positive chronic myeloid leukaemia is presented and compared with the results of peroral chemotherapy with Hydroxyurea or Busulphan in 20 patients. Patients treated with Interferon were significantly younger than the control group (median age 40.5 versus 55.5) (p = 0.01) and were followed-up for shorter period of time (median 10.5 months versus 36.5 months) ( p = 0.002), but did not differ in other parameters. Despite the shorter period of observation and treatment, significantly more complete haematological remissions were achieved with Interferon (86%) than with peroral chemotherapy (25%) (p = 0.03). 6 major and 2 minor (44%) cytogenetic responses were observed after Interferon, despite the fact that 8 patients had been treated for less than one year. Interferon was not the optimal therapy in the patients with additional or complex cytogenetic abnormalities at the time of diagnosis, which were the most significant negative prognostic factor. In general, our short-term results confirm the importance and effectiveness of Interferon in the patient with CML providing the therapy was started early, with an effective dose and with simultaneous cytogenetic monitoring. Longer observation of the patients is needed to confirm the impact of Interferon on the survival of patients. PMID- 9221561 TI - [Fludarabine monophosphate in the treatment of patients with advanced stages of chronic B-cell lymphatic leukemia resistant to conventional chemotherapy]. AB - The authors evaluated retrospectively a group of 12 patients with B-chronic lymphatic leukaemia to whom fludarabine was administered. All patients had been treated in the past by combined chemotherapy (1-4 regimes, median 3). Fludarabine was administered in amounts of 30 mg/m2/day for 5 days to 4 patients and for 3 days to patients. The median of the number of administered cycles was 3. Only two patients achieved partial remission of the disease, the reminder did not respond to therapy. All patients had complications which very probably were associated with the administered treatment. A total of 21 episodes were recorded in the course of 36 cycles, 1 complication per 1.7 cycles. The most frequent complications were infections, a total of 14 episodes, incl. 3 invasive aspergilloses. Infections were more frequent in patients with a 5-day cycle, the majority was recorded after the first two cycles. Eight patients (67%) died from complications which developed in the course of treatment or after its termination. The author's experience with the administration of fludarabine in intensively pretreated patients with advanced forms of B-chronic lymphatic leukaemia indicates that this treatment is associated with a large number of serious complications, which are not compensated by a corresponding therapeutic effect. PMID- 9221562 TI - [The most rapid detection of ischemic strokes]. PMID- 9221563 TI - [Evaluation of diastolic filling of the left ventricle using Doppler echocardiography in patients with chronic ischemic heart disease and with normal left ventricular function]. AB - Twenty-five patients with chronic ischaemic heart disease and intact left ventricular systolic function who had no other cardiovascular, systemic or metabolic diseases were examined by two-dimensional and Doppler echocardiography. At rest and during an isometric load Doppler parameters of left ventricular diastolic filling were obtained such as: peak transmitral flow velocity in early diastole - E, peak transmitral flow velocity in atrial contraction - A, their ratio E/A and deceleration time of early filling. Based on the coronarographic finding, the patients were divided in two ways: 1. group A - 15 patients with critical narrowing of some major coronary artery (stenosis > or = 90% of the luminal diameter) and group B - 10 patients without critical narrowing (stenosis > or = 50%, but less than 90% of the luminal diameter), 2 group C - 6 patients with triple vessel disease, group D - 8 patients with double vessel disease and group E - 11 patients with single vessel disease. None of the groups differed mutually in any of the investigated Doppler parameters of left ventricular filling at rest, nor after an isometric load. CONCLUSION. Diastolic Doppler parameters of left ventricular filling do not make it possible to detect patients with critical coronary narrowing and they do not help to estimate the number of coronary vessels with significant stenoses. PMID- 9221564 TI - [Special features of strokes in diabetics]. AB - Diabetes mellitus is a potent risk factor of cerebrovascular episodes. All statistics reveal a markedly higher prevalence in diabetic patients, as compared with non-diabetics, a poorer prognosis, more frequent recurrence and higher mortality. These facts are particularly relevant with regard to the increasing rate of diabetes in this country and in the world and also with regard to data on the rising incidence of cerebrovascular episodes in diabetic subjects, at least as recorded e.g. in the USA. Prevention is the concern of diabetologists, specialists in internal medicine and all doctors who are in contact with diabetic patients. Essentially it is identical with systematic diabetological care focused on maintaining a normal blood pressure and normal blood lipid values and acceptable blood sugar values in diabetics patients. Principles of secondary prevention must include monitoring of the arteries of the neck by auscultation and instruments and early correction of revealed serious atherosclerotic changes. Unfortunately this is the rule so far. PMID- 9221566 TI - [Treatment of chronic viral hepatitis B and C]. PMID- 9221565 TI - [Treatment of acute cerebrovascular ischemia]. AB - The author presents contemporary views on the main pathophysiological mechanisms of focal cerebral ischaemia and gives an account of possibilities of pharmacological treatment. Consistent with the concept of so-called pharmacological window he divides treatment of acute cerebrovascular ischaemia into treatment of the acute stage (within eight hours after the development of the attack) and treatment of the subacute and chronic stage. He emphasizes the basic importance of early hospital admission to special intensive care units (stroke units) to improve the resulting therapeutic effect and to reduce mortality. He draws attention to the debt of the Czech health services in this sphere of care and treatment of cerebrovascular attacks. PMID- 9221567 TI - [A new purine analog in the treatment of hematologic malignancy. I. Fludarabine]. AB - New purine analogues, fludarabine, 2-chlorodeoxyadenosine and 2-deoxycoformycin are remarkably active in generally incurable malignant lymphoproliferative disorders. The first part of the review summarises pharmacological properties, the mechanism of action, toxicity and clinical use of fludarabine. Major clinical experience with fludarabine has been obtained in patients with chronic lymphocytic leukaemia (CLL). In the studies in pretreated patients with CLL, the overall response rate was over 50%. In previously untreated patients with CLL response rate of 75-80% was recorded with a high ratio of complete responses. Fludarabine was found to be active agent in indolent lymphoma in phase I/II. Approximately 60% of patients with follicular lymphoma respond to fludarabine monotherapy. Combination of fludarabine with cytosine arabinoside (ara-C) is now successfully used in the treatment of acute myelogenous leukaemia (AML) and myelodysplastic syndrome (MDS). Other potential areas of use for fludarabine include hairy-cell leukaemia, Waldenstrom's macroglobulinaemia and mycosis fungoides. Myelosupression, especially leuko and lymphopenia is the major dose limiting adverse effect of fludarabine. A long term reduction in CD4+ T cell count may be associated with an increased incidence of opportunistic infections. Other adverse effects such as nausea and vomiting or neurotoxicity are of mild to moderate severity when the recommended dosage is used. PMID- 9221568 TI - [Peritoneal catheters in the treatment of continuous peritoneal dialysis (CAPD)]. AB - Continual ambulatory dialysis (CAPD) is used during the last five years with increasing frequency in the treatment of patients with chronic renal failure. In some countries already a very high proportion of patients is treated by this method, e.g. in Great Britain about 50%. A decisive influence in the design of peritoneal catheters (PC) and improvement of the exchange system of bags with the dialyzatin fluid. The most important step in the development of the peritoneal catheter was the introduction of a dacron cuff in the intramural portion of the catheter by Tenckhoff. The cuff is a strong barrier against penetration of infection into the abdominal cavity via the subcutaneous tunnel. There are many modifications of the design of the intraperitoneal part, however, they are not of such fundamental importance for the function of the PC as the dacron cuff. As regards implantation of the PC, in addition to the surgical insertion the laparoscopic method is being enforced. A new method is leaving the external portion of the PC in subcutaneous tissue for 3-6 weeks, as recommended by Moncrief and Popovich. This method reduced the incidence of peritonitis after exteriorization of the PC. PMID- 9221569 TI - [The process of ventricular remodeling after acute myocardial infarct associated with left ventricular aneurysm and ventricular septum rupture treated with radical surgery]. AB - Even after a successful operation of mechanical complications on account of acute myocardial infarction gradually developing adverse remodelling of the left ventricle has to be envisaged. In a six-year clinical study by means of echocardiography the authors followed up systematically some cardiac dimensions and volumes and functional systolic and diastolic left ventricular parameters. The changes pertained in particular to the endsystolic and enddiastolic volume, the ejection fraction, the peak maximum rate, early and late diastolic filling and their ratio as well as to indirect values of the mean pressure in the pulmonary artery. These changes, which at first indicated impaired relaxation, are caused subsequently by increasing stiffness of the left ventricle. With regard to the large number of complicated pathophysiological phenomena pertaining to active relaxation and passive elastic properties of the left ventricle during ventricular diastole, different Doppler parameters must be evaluated very carefully, individually and with regard to the clinical condition. Attention is drawn to the importance of complicating mitral regurgitations and an increased pressure in the left atrium and lesser circulation after aneurysmectomy of the left ventricle. Mitral regurgitation has an impact on the process of left ventricular filling investigated by means of diastolic Doppler functions. Despite limitations of echocardiographic methods within the framework of assessment of diastolic left ventricular functions after myocardial infarction echocardiography remains the main means for evaluating left ventricular function by a non-invasive route and its position in this respect is irreplaceable. Further experimental work is needed for better understanding, use and more intelligent interpretation of non-invasive parameters of left ventricular function also in these complicated conditions after surgery of mechanical complications resulting from myocardial infarction. PMID- 9221570 TI - [Ethical problems in molecular genetics and their reflection in clinical medicine]. AB - There are at least two groups of issues connected with an impact of a realization of the "Human Genome Project": a) philosophical; b) ethical, which can be divided into four groups: 1) the influence of DNA technologies on everyday applications of bioethical principles; 2) ethical aspects of genetic diversity; 3) ethical aspects of genetic screening; 4) somatic and germ-cell gene therapy; Unlike essential philosophical issues practical realization of issues in question can be largely expressed as only revitalization of old ones and concerns: the principle of justice-equal access and priorities; protection of reproductive choices; disclosure to patients and to relatives at genetic risk (disclosure and exclusion tests); prenatal diagnosis for "mild to moderate" diseases with and without genetic indication-commercialization; insurance policy; non-directive and directive genetic counseling. Maybe there are regional and other differences, but in practice, the main ethical issues are likely to involve screening for genetic risk of common diseases of adult life e.g. hypertension, diabetes, gout, dyslipoproteinemia, genes for premature atherosclerosis, etc. because of the possible direct impact on a patient, an implication for life-insurance, employers and commercial exploitation. PMID- 9221571 TI - [Clinical experience with changing type I diabetics from animal to human insulin administered by the NovoPen 3 applicator]. AB - The objective of the study was to assess the safety of changing ambulatory patients from animal insulin produced in the Czech Republic administered by classical insulin syringes to human insulins of the Danish firm Novo Nordisk, using a NovoPen 3 applicator. Furthermore antibody levels against hog, bovine and human insulin were assessed. Forty-seven patients with diabetes type I stabilized on an intensified insulin regime were after a four-day preparatory period divided at random into two groups. Patients in group A (n = 22) were after randomization changed to human insulin, patients in group B (n = 25) eight weeks later. From the onset of treatment with human insulins up to the end of the study the mean daily dose of insulin in both groups increased (in group A by 1.51 IU/day, in group 1.35 IU/day). This is not statistically or clinically significant. During the same period a statistically significant decline of the mean value of the daily 8-point glycaemic profile was recorded (in group A by 0.85 mmol/l, in group B by 0.51 mmol/l). Glycosylated haemoglobin declined also significantly in the course of the study (in group A by 1.64%, p = 0.00004, in group B by 1.02%, p = 0.0077). The greatest drop occurred during the preparatory period. Despite the increased daily insulin dose and improved compensation the number of hypoglycaemic events declined significantly in both groups (in group A by 0.78%, p = 0.0102, in group B by 0.74%, p = 0.0134). Hypoglycaemic coma was not recorded in either group. A significant drop of insulin antibodies was found in both group after the onset of treatment with human insulins. From the results of the study ensues that metabolically compensated type I diabetics with a mean daily insulin dose of 0.6 IU/kg body weight can be changed without any complications, in the ambulatory department, to human insulins with the same dosage. Concurrently a gradual decline of antibodies can be expected. PMID- 9221573 TI - [The 104th Internal Medicine Seminar, Bratislava, 17 October 1996. Preoperative preparation of the patient]. PMID- 9221572 TI - [An explanation of the mechanism of attacks of atrial fibrillation in individuals without detectable heart disease]. AB - The authors investigated 10 men aged 30 to 60 years who developed repeatedly attacks of atrial fibrillation. In all patients previous alcohol consumption was recorded in the case-history. In none of the patients organic heart disease was detected. In previous experiments the authors induced by alcohol administration to rats in the tracheal cells and paratracheal myelinated nerves an increased occurrence of lamellar bodies which is consistent with data in the literature which confirmed the presence of lamellar bodies in the human heart muscle in idiopathic cardiomyopathy. The authors assume therefore that alcohol induced increased presence of lamellar bodies in the myocardium which are formed during dystrophic changes, in particular from mitochondria, can lead due to reduced energy release to impaired conduction of excitation in the myocardium. This condition may be enhanced also by impaired calcium metabolism in alcoholic dystrophy. PMID- 9221575 TI - [Selected presentations delivered at the May Hepatology Seminar in Carlsbad on 16 17 May 1996]. PMID- 9221574 TI - [Tumors of the liver and biliary system]. AB - The classification of tumours is based on the traditional classification into epithelial, mesenchymal and less common types. The most frequent epithelial tumour is hepatocellular carcinoma, which usually develops in a cirrhotically altered liver. Its differentiation from regenerative and hyperplastic liver changes by needle biopsy may prove very difficult. Malignant mesenchymal liver tumours are rare. Because of a more frequent incidence and uncertain biological behaviour epitheloid haemangioendotheliomas are more important. Newly described units include also inflammatory pseudotumours and primary MALT lymphoma of the liver. PMID- 9221576 TI - [Etiology and epidemiology of hepatocellular carcinoma]. AB - Hepatocellular carcinoma (HCC) is a highly malignant tumour with a poor prognosis. Its incidence is rising. The estimate incidence worldwide is 1 million cases. Most frequently it develops in livers already affected by cirrhotic transformation. How cirrhosis predisposes for the development of HCC is not clear. It is probably associated with the increased DNA synthesis in regeneration nodules. In micronodular transformation (most frequently alcoholic) the incidence is less frequent than in the macronodular from (mostly posthepatitic). The relationship of HCC and viral hepatitis is beyond doubt-this applies in particular to hepatitis B and C. Chronic alcoholism must not be either. There the risk of development of HCC is four times higher than non-alcoholics. Toxins can be also important for the development of HCC (in particular aflatoxins, chlorinated hydrocarbons, pesticides). As to drugs, in particular anabolics and contraceptives are suspected. Smoking is also a risk factor. HCC is encountered also more frequently in some liver diseases caused by metabolic disorders. It is probable than the development of HCC is a multifactorial process with a marked component of liver transformation. PMID- 9221577 TI - [The clinical picture and diagnosis of liver tumors]. AB - In the diagnosis of hepatic tumour it is most important to differentiate primary carcinoma of the liver from secondary metastatic tumours and focal and infiltrative changes. Primary hepatocellular carcinoma is usually a complication of cirrhosis of the liver. The clinical symptoms therefore correspond especially to an accentuation of the symptomatology of cirrhosis of the liver and complications. The diagnosis is based above all on modern imaging methods such as ultrasonography, computed tomography, nuclear magnetic resonance, examination of tumour markers. Examination by needle biopsy is important and is nowadays usually done under ultrasonographic control to obtain material for histological or cytological examination. Secondary metastases into the liver usually associated with tumours of the pancreas, colon, stomach, bronchi. A frequent diagnostic problem is the recognition of some cysts, capillary, haemangiomas, focal steatosis and focal nodular hyperplasia. An abscess of the liver can also cause differential diagnostic problems. The marked advances in the diagnosis of hepatic tumours are unfortunately not always followed by therapeutic success. PMID- 9221578 TI - [Tumor markers in the diagnosis of tumors of the liver and biliary tract]. AB - Tumour markers are a valuable contribution to the diagnosis and monitoring of tumours processes in the liver. They are used for early diagnosis as well as for monitoring of treatment. In particular monitoring of the dynamic of changes is important. A dominating place is still held by alpha fetoprotein. The importance of CEA is in differentiation of metastatic processes. In the diagnosis of neoplastic processes in the biliary pathways antigen CA 19-9 is most important. An important finding is also than non-specific increases of tumours markers occur in some chronic processes of the liver and biliary pathways. Despite the large number of tumour markers they are useful only in conjunction with other clinical and imaging methods. PMID- 9221579 TI - [Regional chemotherapy of liver tumors]. AB - Based on experience with regional intrahepatic chemotherapy in 67 patients with inoperable primary, and in particular secondary liver tumours, which comprises several hundred administered cycles of mostly continuous regional regimens, the authors summarize briefly the main principles and possibilities of this treatment. In a review of randomized studies they provide evidence for the advantage of regional chemotherapy, as compared with systemic treatment, in particular with regard to the higher percentage of therapeutic responses. In correctly indicated, and if possible early cases of hepatic tumourous affections, it is possible to potentiate the effect of regional chemotherapy by local destruction of tumourous foci by alcoholization, cryodestruction or resection. An integral part of this treatment is also monitoring of the effect by following up the dynamics of serum levels of tumour markers and by imaging methods. Because the most frequent cause of failure of this method are extrahepatic secondaries and secondary chemoresistance of the tumourous foci, improvement of results can be expected in particular from a combination of regional and systemic chemotherapy and the inclusion of cytokines into the therapeutic schemes. PMID- 9221580 TI - [Cytokines in regional immunochemotherapy of liver tumors]. AB - Regional intrahepatic chemotherapy of inoperable primary and secondary liver tumours can achieve, as compared with the little effective systemic chemotherapy, a higher percentage of therapeutic responses. The objective of regional chemoimmunotherapy, i.e. the use of cytokines, in particular interferon alpha (IFN-a) and interleukin-2 (IL-2) in the therapeutic regimens is to improve the survival of patients with malignant liver tumours. One of the main prerequisites of the effect of locally administered cytokines is activation of hepatic lymphocytes (LAL)-liver associated lymphocytes, effectors with specific phenotype and potential anti-tumourous effect directly in the target area. Although in regional monotherapy the effectiveness of cytokines is low, regimens combining the administration of cytostatics with IL-2 achieve a 50-70% therapeutic response. The authors summarize basic data on the regional administration of cytokines and present an review of combined regimens of regional chemoimmunotherapy, including their own protocol of the Masaryk Oncological Institute in Brno. PMID- 9221581 TI - [Transhepatic therapy of tumors of the biliary system]. AB - The transhepatic percutaneous approach is a supplementary diagnostic and therapeutic method in case of biliary obstruction, in particular when the endoscopic approach fails. The bile ducts are punctured by a thin needle and then drained, followed if desired by insertion of a prosthesis. Although the method is associated with more complications than standard endoscopy, it markedly improves therapeutic possibilities. PMID- 9221582 TI - [Treatment of liver tumors with ethanol injection]. AB - Ethanol injection into the focus by means of a thin needle under ultrasonographic guidance has become part of treatment of primary carcinoma of the liver hepatocellular carcinoma. It is a palliative method suitable for the treatment of small foci of hepatocellular carcinoma in a uniocular or oligolocular form. The limiting factor are in particular the number of foci, their size, depth and stage of hepatic cirrhosis in the remainder of the live parenchyma. Treatment is well tolerated and the survival of patients, if they are properly selected, is similar as in surgical resection. PMID- 9221583 TI - [Epidemiology and etiology of tumors of the biliary tract]. AB - A review of etiological factors participating in the development of tumours of the biliary pathways. Epidemiological data on the incidence of tumours of the biliary pathways in different part of the world are presented. PMID- 9221584 TI - [Intraluminal brachytherapy in tumors of the biliary system]. AB - The therapeutic possibilities in tumour of the biliary pathways are rather limited and the survival of patients treated by palliative methods is usually short. The most frequent complication of tumours of the biliary tract is biliary obstruction. A palliative treatment involves surgical derivation of bile the endoscopic or percutaneous route. Irradiation as part of palliative therapy is limited by the relatively low sensitivity of the tumours to radiation and the close contact with various radiosensitive organs. One of the possibilities how to avoid damage of neighbouring organs is local actinotherapy. A suitable approach for introducing a radiation source is a percutaneous drain in the biliary tract. There are several patterns of irradiation programmes, most frequently Irridium 192 is used. The total dose of 30 Gy is divided into six fractions which are administered in the course of 2-3 weeks. Local irradiation is followed by implantation of a metallic self-expanding stent into the stenotic part. The effect of this treatment is diminution of the tumours mass, patency of the branches of the intrahepatic bile ducts obstructed by the tumour and prolongation of the patency of the implanted stents. Complications of local actinotherapy are minimal, the majority of complications is associated with percutaneous drainage. The most suitable indication for this treatment are non-resectable tumours of the upper part of the biliary tract, less suitable are tumours of the gallbladder and ampullomas of the papilla Vateri. In the above indication local actinotherapy is palliative treatment which prolongs survival and improves the patient's comfort. PMID- 9221585 TI - [An analysis of the reasons for an unjustified call-up for military service based on health status]. PMID- 9221587 TI - [Experience with the hygienic analysis of the causes and circumstances of the occurrence of trauma with a fatal outcome in the operations of military engineering installations]. PMID- 9221586 TI - [New problems of military medical geography]. AB - The new requirements to the military medico-geographical researches, to contents and structure of military medico-geographical descriptions are considered, caused by evolution of global geopolitical situation, by occurrence of the modern information technologies, the geographical information systems, by development of the medical ecology and information medical service. Principles of the concept of the ecological military-medical geography are proved. The priority problems and specific peculiarities of the units are stated. It is recommended wider use of the factor's approach and cartographic method to enhance the organization of medico-ecological maintenance of the Army and the Fleet personnel activity. PMID- 9221588 TI - [The modern view on the problem of combat-related mental trauma]. PMID- 9221589 TI - [The diagnosis of damages to the auditory system in the early period of explosive mine trauma and the optimization of treatment for the victims]. AB - The wide use of the mine-explosive weapon in modern military conflicts is accompanied by high specific weight of mine-explosive traumas (MET), which achieve 30-40% of all traumas. In the departments of the hospital in 21.3-35.8% of the wounded with MET accompanying defeat of acoustical system with probable subsequent development of neurosensory hypoacusis and invalidism of the injured is established. It defines necessity of early ENT-inspections of all injured with such traumas. Steady up to 1-6 months reduction of hearing is accompanied by biochemical infringements of a cerebrospinal liquid. PMID- 9221590 TI - [The level of and outlook for the development of specialized medical care in the department of maxillofacial surgery and stomatology of a central hospital]. PMID- 9221591 TI - [Color Doppler mapping using power Doppler in assessing prostatic pathology]. PMID- 9221592 TI - [The comparative efficacy of Aurorix and amitriptyline in treating depressions]. PMID- 9221593 TI - [The anti-anginal efficacy of the drug forms of isosorbide-5-mononitrate- Monocinque, Monocinque retard and isosorbide dinitrate]. PMID- 9221594 TI - [Methodological bases for using computer technologies in the activities of the medical service of the Armed Forces]. AB - Authors state methodological bases of construction of automated control system (ACS) of the medical service of the Armed Forces and the Navy. A basis of the methodological rules is made by the analysis of conformity of the purpose (preservation and strengthening of the servicemen' health and management composed process: the tax, processing, storage and distribution of data and preparation of the decision, i.e. the information-and-target approach. The automation of processes of profile activity of forces and means of the medical service provides the constant control by functioning of medical units and institutions. The main targets of the ACS' functioning are processing of the information about quality of rendering of the medical care to the servicemen, its timeliness and profitability, productivity of their labour, conformity of work results to the medical standards and others. The quality and the efficiency of ACS'application is defined by a level of information maintenance. ACS of the medical service should be subsystem of the automated control system of the troops. PMID- 9221595 TI - [Methodological approaches to the study and prevention of shigellosis among the troops]. AB - In article the methodical approaches to study of regularity of occurrence and distribution of shigellosis are revealed from positions of the theory of etiologic selectivity of main ways of transfers, its unequivalentness at the various nosologic forms of dysentery. A complex of antiepidemiologic measures under conditions of means of prophylaxis is proved. On the basis of analysis of etiologic structure of shigellosis a main problem of the Armed Forces, causing dysentery is designated. PMID- 9221596 TI - [The immuno-microbiological prediction of a protracted course in Sonne dysentery]. PMID- 9221597 TI - [Adaptation and military professional training]. PMID- 9221598 TI - [A method for assessing the medical technology level of mobile medical units]. PMID- 9221599 TI - [The start of a new age in Russian surgery (on the 150th anniversary of the first use of anesthesia in Russia)]. PMID- 9221600 TI - [The flagship of aerospace medicine (on the 55th anniversary of the Central Military Research Aviation Hospital of the Ministry of Defense of the Russian Federation)]. PMID- 9221601 TI - [Current problems in the correction of functional states and rehabilitation in aerospace medicine]. PMID- 9221602 TI - ["Brother of sleep". The operating room and temperature]. PMID- 9221603 TI - [Intraoperative hypothermia: pathophysiology and clinical sequelae]. AB - Both regional and general anesthesia markedly impair the normal precise regulation of core body temperature. Consequently, inadvertent perioperative hypothermia is common. Hypothermia develops because the typical operating room environment is cold; however it is anesthetic-induced impairment of thermoregulatory responses that contributes most. Internal redistribution of body heat is a surprisingly important factor, contributing more to core hypothermia than net heat loss in most patients. There is now convincing evidence that a typical degree of intraoperative hypothermia, say 2 degrees C, predisposes to numerous complications such as shivering, prolonged duration of action of several drugs, myocardial ischemia, coagulopathy and increased incidence of surgical wound infections, which alter patient outcome. Fortunately, effective methods such as convective warming are available for preventing hypothermia. PMID- 9221604 TI - [Annual treatment rates and estimated incidence of eating disorders in Austria]. AB - BACKGROUND: At present, the prevalence and incidence of eating disorders in Austria is unknown; not even rough estimates of countrywide annual treatment rates are available. AIMS: To assess the number of patients in Austria with eating disorders currently under treatment and to compare this rate with the estimated prevalence and incidence of eating disorders, thus providing an estimate of unrecorded cases and the appropriateness of health care for these disorders. METHODS: The number of patients being treated in major out-patient and in-patient facilities was assessed by questionnaires. Prevalence and incidence rates in Austria were estimated by extrapolation of epidemiological data from comparable Western countries to Austrian figures from the most recent population census in 1991. RESULTS: Altogether 1075 patients were being treated in 1994 at 26 institutions, including all specialized centres, pediatric and psychiatric university hospitals. Surprisingly, the annual treatment rates for anorexia and bulimia nervosa were equal. There is a considerable discrepancy between these treatment figures and prevalence/incidence estimates (in absolute numbers): the estimated anorexia nervosa point prevalence is about 2500 girls aged 15-20 years, whilst a minimum of 4400 girls suffer from subclinical eating disorders, and there are about 6500 bulimia nervosa cases in young women aged 20-30 years. The incidence might be about 600 new onset cases per year for anorexia, and about 870 for bulimia nervosa. The size of the problem (lifetime prevalence) may comprise at least 36,000 women with bulimia nervosa. CONCLUSIONS: Eating disorders pose a major public health problem for women in Austria. It is unlikely that the vast majority of unrecorded cases was treated in private practice or in hospitals which failed to respond to our questionnaire. The discrepancy between annual treatment rates and prevalence/incidence estimates points to a lack of specialized eating disorder units in Austria. PMID- 9221606 TI - [Julius Tandler and the Vienna-Lainz City Hospital]. PMID- 9221605 TI - [Hyperthyroidism caused by TSH producing hypophyseal adenoma]. AB - Thyrotropin (TSH-)producing adenomas of the anterior pituitary gland are the least frequently encountered ones and constitute a very rare cause of hyperthyroidism. The case is presented of a 58 year old male patient with a well known history of hyperthyroidism over a period of at least 9 years growing goiter. Despite different forms of medical treatment he presented a constant clinical pattern consisting of restlessness and paroxysmal tachycardial atrial fibrillation. Laboratory findings revealed elevated levels of circulating thyroid hormones despite inadequately high levels of TSH. MRI scan revealed an adenoma of the pituitary measuring 9 mm in diameter. After microsurgery, consisting of transphenoidal resection of the tumor, the patient recorded no clinical symptoms. Histological examination revealed positive immunohistochemical staining, with antibodies to TSH, but a negative reaction against the GH, PRL, FSH, LH and ACTH hormone antibodies. Moreover, the levels of circulating hormones (GH, PRL, FSH, LH and ACTH) were normal. TSH-alpha subunits were not elevated. Before the correct diagnosis was reached, this patient was treated for nine years with antithyroid drugs. Five months after the operation the patient showed normal values of circulating thyroid hormones and TSH and thus no thyroid-specific medication was necessary. PMID- 9221607 TI - [Professor Julius Tandler. In memory of and comments in honor of the 60th anniversary of his death]. PMID- 9221608 TI - [Preparing for colonoscopy: a reliable and easily implemented regimen]. PMID- 9221609 TI - [1997 Paul Ehrlich and Ludwig Darmstaedter Prize for the rediscovery of Helicobacter pylori to J. Robin Warren and Barry J. Marshall]. PMID- 9221610 TI - [Phospholipase A2--acute phase protein after liver surgery?]. AB - INTRODUCTION: The exact source of phospholipase A2 is unknown. In this context the liver is discussed because the secretion of phospholipase A2 could be initiated during an acute phase response. PATIENTS AND METHODS: Three groups of patients were prospectively established: A liver resection (n = 12); B control (n = 22; oesophageal resection, gastrectomy, rectum resection); C sepsis (n = 5). Blood was collected preoperatively, each day postoperatively for seven days and at the day of discharge. Biochemical procedures: Phospholipase A2, PMN-elastase, C-reactive protein, GPT, GOT, GLDH, cholinesterase. RESULTS: Postoperatively phospholipase A2 is unchanged in the liver resection and control group, whereas it increases significantly in septic patients. Only following liver resection GPT, GOT and GLDH increase. C-reactive protein is increased in all groups. DISCUSSION: Unchanged phospholipase A2 after liver surgery despite an acute phase response indicated by C-reactive protein does not support the view of phospholipase A2 being of hepatic origin. PMID- 9221611 TI - [Occlusion of an esophagobronchial fistula by implantation of a Montgomery esophageal and a dynamic tracheal stent after failure of conventional endoprosthesis]. AB - Esophagorespiratory fistulas were frequently caused by malignant tumors, bougienage, laser therapy or radiochemotherapy. We here report the case of a patient with inoperable bronchial cancer, who developed a symptomatic esophagorespiratory fistula during combined radiochemotherapy with Cisplatin. A sufficient occlusion of the fistula could not be achieved with conventional plastic tubes or novel self-expanding silicone-coated Gianturco Song stents. After extraction of two Gianturco Song stents we inserted a Montgomery Salivary Bypass Stent into the esophagus and Dynamic stent into the trachea. This resulted in a total occlusion of the fistula. This present case suggests that the Montgomery stent may have little tendency to migrate due to its characteristic configuration and fixation and further demonstrates that the novel self-expanding silicone-coated Gianturco Song stents can be removed, if necessary. PMID- 9221612 TI - [The intestinal hormone glucagon-like peptide 1 (GLP-1): from experiment to the clinic]. AB - A functional connection between the small intestine and endocrine pancreas was proved in the sixties, after it became possible to determine the exact amount of insulin in plasma. The insulin response after oral doses of glucose is substantially stronger than after intravenous doses of sugar, even when identical glucose plasma levels are attained. This incretin effect is explained by the connection of the entero-insular axis. The intestinal hormones, that are released by the small intestine after meals, circulate measurably in plasma, and strengthen the glucose-induced insulin secretion, are responsible for this effect. In addition to the classical incretin hormone "Gastric inhibitory polypeptide-1" (GIP), "Glucagon-like peptide-1" (GLP-1) is very interesting to investigators today. In a relatively short amount of time, GLP-1 has matured from a physiologically interesting incretin hormone candidate to a potentially therapeutical alternative for the treatment of diabetes mellitus. GLP-1 stimulates glucose-dependent insulin secretion, decreases plasma glucagon levels, delays gastric emptying, and putatively exerts an additional effect on peripheral glucose utilization. On top of that, GLP-1 has effects on the central nervous system thereby impacting on feeding behavior. PMID- 9221613 TI - [Experimental methods in hepatology. Guidelines of the German Work Group for Study of the Liver. Therapy of ascites in liver diseases. German Work Group for Study of the Liver]. AB - Patients with "uncomplicated" ascites (no encephalopathy, electrolyte derangements or markedly impaired renal function) respond well to the conventional sequential therapy comprising dietary sodium restriction, aldosterone antagonists and loop diuretics. A close monitoring of the patients is important to avoid side effects. For massive ascites, ascites refractory to diuretic treatment or recidivant ascites daily therapeutic paracentesis is recommended. Four to six liters of ascites should be drained daily followed by infusion of plasma expanders (preferentially albumin, eventually hemacell or dextran-70) until the patient is almost free of ascites. The diuretic treatment should be instituted. The transjugular intrahepatic portosystemic shunt (TIPS) represents a promising novel therapeutic modality the therapeutic impact of which has to be more clearly defined in prospective studies. Several experimental approaches have been proposed for the therapy of the hepatorenal syndrome (Dopamine, Ornipressin, TIPS). Liver transplantation is the only causal treatment of refractory ascites or hepatorenal syndrome and should be discussed when a patient presents with massive ascites. PMID- 9221614 TI - [Decompression of the main pancreatic duct can delay progressive loss of pancreatic function in chronic pancreatitis]. PMID- 9221615 TI - [Intestinal cell proliferation, acromegaly and colon tumors]. PMID- 9221616 TI - [The "diabetes" (db) gene of the mouse codes for a mutated leptin receptor--a cue to understanding obesity?]. PMID- 9221617 TI - [Isoprostanes, a new substance group in angiology--of future significance?]. AB - F2-isoprostanes are prostaglandin F2-like compounds being formed by non-enzymatic peroxidation of arachidonic acid in vivo. They have a variety of biological actions. The most important compound of this group is 8-epi-PGF(2 alpha) being capable to induce vasconstriction in particular of lung- and renal vascular tissue. Isoprostanes are present in esterified form; in free form they become available after hydrolysis by phospholipase A. An increase in isoprostanes is an important indicator of oxidative stress in-vivo due to a variety of different noxi such as metal- or non-metal ions for cigarette smoke. Isoprostanes show an activation of platelets; as a consequence of the interaction of 8-epi-PGF(2 alpha) with specific receptors platelet aggregation may be induced or may be enhanced together with other agonists. Due to these preliminary results isoprostanes could become an interesting substance in angiology in the future for diagnosis of oxidative stress as well as in the understanding of the pathogenesis of atherosclerosis. PMID- 9221618 TI - [Vaginal speculum: the developmental history of a gynecologic instrument]. AB - The vaginal speculum (katopter) was known in Antiquity. The instrument fell in disuse during the Middle-ages notwithstanding the fact that Abulcasis had conceived improved speculae in the 10th century. Obstetrical use (embryotomy) of the speculum matricis was deleted from the 17th century on. The speculum vaginae was "rediscovered" in 1801. PMID- 9221619 TI - [Juvenile hormones, reality or myth?]. AB - Recently, the "discovery" of so called "rejuvenating" pills, has attracted much interest in the media and has raised irrational expectations among the elderly population. These so called "rejuvenating drugs" are dehydroepiandrosterone (DHEA), a steroid hormone secreted by the adrenal cortex, and melatonin, an indol derivative, secreted by the pineal gland, both known since many years. Dehydroepiandrosterone is quantitatively by far the most important steroid in the human organism; it is a weak androgen, a small fraction of which is aromatized to estrogens in peripheral tissues. Plasma levels of DHEA decrease with age and some authors consider these levels as a reliable parameters of biological age, the more so that some studies seem to indicate that low levels are accompanied by increased morbidity and mortality. This could not be confirmed, however, by other authors. In in vitro experiments, DHEA has anti-oxidative effects, inhibits platelet aggregation and, possibly, stimulates the immunological system. In animal experiments DHEA has some antitumoral effects. These effects were, however, observed in animal species which do not secrete DHEA. It should, moreover, be mentioned that administration of DHEA in a high dosage, induced the development of hepatic carcinoma in 14 out of 16 rats. Preliminary controlled studies, performed in 30 elderly persons, showed an improvement of general wellbeing with an increase in plasma IGF-1 levels; in women a moderate increase in plasma testosterone and estradiol was observed. Hence these studies show a moderately beneficial effect of DHEA therapy. They need to be confirmed and they warrant further well controlled studies. Melatonin is a hormone secreted by the pineal gland, the releasing stimulus of which is darkness. It synchronizes the biological rhythms as well as the seasonal biological changes induced by the photoperiod. It induces sleep and is an euphoretic. Melatonin plasma concentrations decrease with age and this decrease has been related to the impaired sleep induction in elderly. Melatonin levels are also decreased in depression. In mice, transplantation of the pineal gland of young animals to old animals increases life expectancy by +/-20%. In in vitro experiments, melatonin appears to have some anti-oxidative effects, which led to the hypothesis that it might retard the ageing process and inhibit the growth of tumor cells. In man, melatonin has been shown to be effective in preventing jet lag and in improving sleep induction in the elderly as well as disturbances of the nycthemeral rhythms in blind persons. One research group even reported favourable effects of a combined melatonin-IGF-1 treatment of metastatic carcinomas; these results were not confirmed up to now. As to the advertised effects on sexuality, it is well known that melatonin inhibits gonadotropin secretion, which causes gonadal atrophy. Hence it is evident that we can not expect a stimulation of sexuality by melatonin administration. As to prolongation of life expectancy, there are, so far, no indications for such an effect in man. PMID- 9221620 TI - [Image-guided surgery]. AB - Initially, stereotactic surgery was developed to treat functional brain diseases only. The localisation of targets was based on stereotactic atlases and radiographs. The introduction of computer based imaging techniques, such as CT and MRI, have offered the possibility to "see" anomalies and to approach them stereotactically. The working principle of such a procedure consists of three steps. It is assumed that brain tissue does not move with respect to the skull. 1. Acquisition of the images and their registration with the patient, usually based on a series of reference points (fiducials) that belong to a stereotactic localizer attached to the base ring of the stereotactic frame, 2. planning and simulation of the surgical intervention, mostly based on two-dimensional (2D) images resliced along arbitrary directions, and 3. intra-operative guidance of the instruments mounted onto the stereotactic frame. This procedure has continuously been updated by new image acquisition techniques, 3D visualization and frameless stereotaxy. 1. PET, MRI and angiography (DSA, MRA) have been used in addition to CT. Moreover, such images of different modalities can automatically be fused without using a stereotactic frame or other artificial fiducials. 2. 3D images for surgery planning have become available, a feature that has proved to be very useful for the cerebral blood vessels. 3. The stereotactic frame can be replaced by a robot arm or an optical guidance system. Registration of the instruments with the patient is then performed by using markers in the cranial bone or on the scalp, or by means of intra-operative images such as radiographs or video images. Recently the use of registered video images has resulted in a number of experiments with "improved reality" and telesurgery. The same working principle has shown to be useful for bone and bone related surgery. As in brain surgery, the prerequisites of rigidity and immobility with respect to a reference are satisfied. Because bone structures are rigid and can easily be outlined in CT images, 3D graphical as well as stereolithographic representations can be produced for the purpose of planning and even for navigation. Unfortunately, for most organs or soft tissue the above conditions of inflexibility and fixed position with respect to a reference are not fulfilled. Real time imaging, partly due to the introduction of the "open" MR scanner, may offer a solution. It can be expected that interventional diagnostic imaging will become increasingly important in the future, also for neurosurgery. PMID- 9221621 TI - [Dermatology in current medicine]. AB - Dermatology evolved in the second part of the last century as a branch of internal medicine and was for many years confined to morphologic descriptions. Skin diseases are common and vary enormously in severity. Although most of the conditions are not life threatening, many of them are debilitating due to functional loss, pain and itch, and the social problems they cause. Skin diseases are a major public health problem in developing countries. After the second world war there has been an explosion both in the amount of scientific knowledge and with regard to the treatment of dermatoses, many of which can now be managed adequately. The dermatologist is also involved in prevention of skin diseases, as e.g. contact dermatitis and malignant tumors of the skin. Increasing specialisation within dermatology has become more common with the expansion of expertise in dermatopathology, photodermatology, contact dermatology, dermatological surgery, dermatologic pediatrics, phlebology. There are too many dermatologists in our country. If an increasing role in caring for minor skin diseases would be assigned to the generalist, the government he will have to decide how contraction of dermatology manpower would occur and how the dermatology training of the generalist should be improved. PMID- 9221622 TI - XXIInd Meeting of the European Tumor Virus Group. Innsbruck-Igs, March 5-9, 1997. Abstracts. PMID- 9221623 TI - [Increasing use of heroine and cocaine in Switzerland since 1990: use of a generalized Poisson distribution in the collected data]. AB - Estimates of the prevalence of deviant behaviour, which are based on the usual survey methods are by far too low. Therefore, the use of capture-recapture methods or of the truncated Poisson distribution is to be preferred, provided appropriate data are available. Here an extended Poisson approach was applied in order to estimate the number of users of hard illegal drugs (heroin, cocaine) in Switzerland for each year from 1990 to 1993. These estimates indicate an increase by about 50% during the period 1990-1993. PMID- 9221624 TI - [Cost-effectiveness analysis of prevention of reinfarction using low-dose acetylsalicylic acid; model calculation]. AB - The purpose of this study is to estimate the potential of savings which can be achieved by prophylaxis of myocardial reinfarction with low-dose acetylsalicylic acid (ASA) at 75 mg per day over a treatment period of two years. After secondary analysis of published data, the effectiveness of low-dose ASA is compared to placebo by a model calculation. The difference in the effectiveness between the prophylaxis with ASA and placebo is taken from an international meta-analysis. The economic valuation of this difference is carried out by a cost-effectiveness analysis applying disease costs per case. According to the model calculation, 5535 DM can be saved per patient with a history of myocardial infarction with 75 mg ASA a day over a treatment period of two years. In 1991 there were around 740,000 patients with a history of myocardial infarction in the age group of 25 64 in the Old Bundeslander of the Federal Republic of Germany. The application of the results of the model calculation would lead to considerable savings. Even in the sensitivity analysis with different assumptions regarding costs incurred by hospital treatment and costs incurred by premature retirement, the cost advantage of the ASA-prophylaxis remains. Due to the cautious and conservative assumptions in the model calculation the potential of savings is likely underestimated. Nevertheless, there is a distinct advantage for the prophylaxis with low-dose ASA which already occurs in direct costs thus leading to advantages also for cost carriers. PMID- 9221625 TI - [Prone sleeping position and other risk factors in sudden infant death syndrome: a prevalence study in Geneva]. AB - A survey by telephone interviews has been carried out in 1993 in the canton of Geneva, in order to measure the prevalence of 3 risk factors for the sudden infant death syndrome (SIDS) and to evaluate the potential for SIDS prevention based on these factors: prone sleeping position, tobacco smoking by the parents during pregnancy, and no complete breast-feeding at 8 days of age. 278 families participated to the study, of 320 families who could be contacted by telephone in Geneva, from a random sample of 550 families having had a child in Geneva in the preceding 12 months. 40% of the infants had been put to bed in the prone position on the preceding evening 18% of the mothers had smoked during pregnancy. Prevalence of low birth weight was 4.9% for 0 to 4 cigarettes per day during pregnancy, 17.2% for 5 cigarettes and more. At one week, 16.5% of children were not exclusively breast-fed. On the basis of these results, it can be estimated that an effective prevention programme, centered on prone sleeping position could decrease the incidence of SIDS by 50% or more. An even greater fraction of cases, up to 80%, could be avoided by the prevention of the 3 studied factors. These estimates show the need to develop a mother and child health programme in Switzerland. PMID- 9221626 TI - [Bias due to non-responders in an epidemiological study (SAPALDIA)]. AB - Within the Swiss Study on Air Pollution and Lung Diseases in Adults (SAPALDIA) 16267 adults aged 18 to 60 years from 8 different locations in Switzerland were randomly selected for answering a questionnaire about respiratory health and have a lung function examination with allergy test. 9561 subjects agreed with the examination (59%) (= responders, R). In order to study the possible influence of the bias introduced by non-responders (NR), 221 subjects who refused to participate among the 966 first subjects selected in Payerne were contacted by phone. 142 accepted a home visit and answered a shortened questionnaire about the main respiratory symptoms and diseases and indicated furthermore the reasons for their refusal. Non-responders have a lower mean educational level and belong to lower social classes than responders. The frequency of respiratory symptoms and diseases, allergies and smoking is similar in R and NR except a higher frequency of wheezing during the last 12 month (R: 12.5%, NR: 5.6%, p = 0.03). The level of carbon monoxide in expired air is higher in NR (17.6 ppm) that in R (11.9 ppm) (p = 0.01). A similar difference exists between NR (30.7 pp) and R (24.8 ppm) among current smokers (p < 0.01). The main reasons for refusal are lack of time (27.5%), lack of interest for medical study (22.6%), fear of health professionals (18.3%) or the existence of a another disease (9.9%). Furthermore, 2.8% of the subjects consider a medical study as useless and refuse principally any participation. The role of local press and media in the decision to participate seems to be important. Globally, the differences between R and NR are minimal and should not influence the validity of the results of the SAPALDIA study. PMID- 9221627 TI - [Limitations of meta-analysis from published data in epidemiological research]. AB - Metaanalyses of epidemiological studies have increased during the last years and are often used to evaluate the effect of risk factors which are inconsistent in different studies, mainly for small risk factors. Very often a metaanalysis is performed from published data. In this article we discuss this form of a metaanalysis and investigate whether the requirement to get reliable information is achievable with it. We mainly ask questions whether qualitative and quantitative dose-response analysis can be performed. We point out the differences between metaanalysis from experimental data and clinical randomized studies and epidemiological studies. We discuss different arguments that were given for performing metaanalysis in clinical trials and investigate whether they are also valid in observational studies. We mainly concentrate on the problem of estimating a single pooled risk estimate. Two examples from literature are used to show problems with metaanalysis from published data. PMID- 9221628 TI - [Interdisciplinary treatment of thyroid diseases]. PMID- 9221629 TI - [Dissent over standards in adjuvant management of colon carcinoma]. AB - Since 5-fluorouracil (5-FU) plus levamisole substantially reduces the recurrence rate and improves survival in adjuvantly treated patients with curative resected stage III colon cancer this combination has been considered the standard therapy. Shortly thereafter folinic acid modulated 5-FU-therapy also demonstrated adjuvant efficacy compared to surgery alone. Therefore various schedules of folinic acid modulated 5-FU-therapy were compared with the standard regimen (5-FU/levamisole). Randomized multicentric studies revealed: In three of four studies 5-FU/FA is superior to 5-FU/levamisole. Treatment duration of 6 months for 5-FU/FA is similar to 12 months 5-FU/Levamisole. No benefit is obtained for 5-FU/FA by additional levamisole. The effect of regional (portal vein or intraperitoneal) treatment is controversial discussed, but combination of the treatment with systemic chemotherapy versus 5-FU/levamisole demonstrated slightly increased therapeutic efficacy. Immunotherapy with autologous tumor cell-BCG or monoclonal antibody treatment improved survival and is currently investigated in studies with conventional systemic treatment and combined chemoimmunotherapy. Beside treatment in studies patients with colon cancer stage III should be offered adjuvant chemotherapy with 5-FU/FA. Further improvements and adjuvant treatment protocols for stage II carcinoma have to be investigated in studies. PMID- 9221630 TI - [Basedow disease hyperthyroidism--is there a therapeutic standard? Conservative therapy]. AB - In patients with the first manifestation of hyperthyroidism of Graves' disease, antithyroid drug treatment is the therapy of first choice. Treatment has to be carried out depending on iodine supply of the individual patient with the lowest possible drug dose. Controls of treatment have to be done in short intervals (every 2 weeks) until euthyroidism is reached, afterwards controls of thyroid function have to be done every three months. After euthyroidism is established, the combination of antithyroid drug therapy with thyroid hormones may be useful to avoid hypothyroidism or goiter development during treatment in contrast to a monotherapy with antithyroid drugs. Antithyroid drug treatment has to be carried out for one year. The remission rate of patients does not increase with higher doses of antithyroid drugs or a longer treatment duration. The determination of TSH receptor antibodies does not help predicting a relapse of hyperthyroidism of Graves' disease in the individual patient at the end of treatment. Regular follow up controls after antithyroid drug treatment are necessary to recognize relapse of Graves' disease in time. PMID- 9221631 TI - [Radioiodine therapy of Basedow's disease--is there a therapeutic standard?]. AB - This review article will come to a positive answer to the question entitled. The good experiences within more than 50 years were able to define commonly accepted standards concerning the following items: therapeutic aim, indications, contraindications, pretreatment, dose concept, assessment of radioactivity, follow-up. The broad consensus among experts when to apply medical treatment, surgery or radioiodine in this special disease was one of the reasons that the nuclear medicine treatment is accepted more and more. By this, the patients have to wait for many months to become treated on an in-patient basis, as it is mandatory in Germany. In the current health care scene it is absolutely unrealistic to demand an increasing number of radioiodine beds. Facing this fact the guidelines of applying radioiodine which are extremely restrictive in Germany have to be modified as proposed by the National Commission on Radiation Protection. PMID- 9221632 TI - [Basedow's disease--thyroidectomy or subtotal resection?]. AB - Postoperative complications and long-term results were retrospectively investigated after near total thyroidectomy due to Graves' disease in 73 patients. Postoperatively one permanent recurrent nerve palsy and one hypoparathyroidism were seen. Two patients had a postoperative bleeding and one patient had a wound infection. No patient died. After a median follow up time of 67 months 65 patients (89%) were seen. A recurrent hyperthyroidism developed 3% of the patients and 80% were hypothyroid. These results demonstrate the effectiveness of near total thyroidectomy in Graves' disease. With no mortality and low morbidity excellent long-term results could be achieved. PMID- 9221633 TI - [Risk factors and follow-up of recurrent laryngeal nerve paralysis after first surgeries of benign thyroid diseases. Results of a retrospective analysis of 1,556 patients]. AB - PATIENTS AND METHODS: risk factors of recurrent laryngeal nerve (RLN) palsy after thyroid gland surgery were evaluated retrospectively in 1556 patients who were submitted to an operation because of a benign thyroid disease. Recurrences were also excluded. RESULTS: RLN palsy occurred in 6.6%. In relation to the nerves at risk the incidence of primary postoperative nerve damages was 4.3%. After a long term follow-up of in total 18 months the incidence of permanent nerve palsy was 1.6% (related to the nerves at risk: 1.1%) as 75.5% of the paralyses were transient in an average of 6.2 months. Substernal goitres especially when sternotomy became necessary, the ligature of the inferior laryngeal artery, serious perioperative complications and total lobectomy in comparison to subtotal resection were important risk factors for primary postoperative RLN palsy (p < 0.05 resp. p < 0.01). The ligature of the inferior laryngeal artery and the extension of resection were indeed significant risk factors also for permanent nerve damages, but the other factors had no influence on the risk of permanent RLN palsy. However, the non-exposure of RLN in subtotal lobectomy was significantly associated (p < 0.01) with permanent, but not with transient nerve palsy. CONCLUSION: The exposure of the RLN is one of the most important procedures during thyroid surgery and particular also during subtotal lobectomy to reduce the rate of permanent RLN damages. PMID- 9221634 TI - [Clinical aspects and preoperative diagnosis in differentiated thyroid gland carcinoma]. AB - Etiologic factors such as iodine deficiency and exposure to ionizing radiation especially during childhood are determinants of histological type and malignancy of differentiated thyroid carcinomas. The rarely occurring anaplastic thyroid carcinoma represents a highly malignant tumour of old patients, causing local compression symptoms at an early stage of disorder. Since early symptoms are lacking, sensitivity and specificity of further diagnostic approaches are of particular importance in the care of differentiated thyroid carcinomas. Obligatory procedures at the preoperative stage are ultrasound, scintigraphy, determinations of thyroid hormones and calcium on serum as well as an otolaryngological investigation. The preoperative determination of calcitonin may assist in the differential diagnosis of a cold nodule and in the detection of a medullary thyroid carcinoma. Measurements of thyroglobulin are helpful only for the follow-up of patients with differentiated carcinomas. The highest degree of specificity and sensitivity is achieved by fine-needle aspiration. Finally, if clinical suspicion of malignancy exists, surgical intervention should be performed even if the result of cytological analysis was negative. Whether the color Doppler energy ultrasound provides valuable additional informations especially in the differential diagnosis of thyroid nodules during childhood, is still undecided. PMID- 9221635 TI - [Surgical therapy of differentiated thyroid gland carcinoma]. AB - Surgical excision is the treatment of choice for differentiated thyroid carcinoma. Because no prospective randomized clinical trials are available, decisions regarding the extent of surgical resection and the mode of lymph node dissection must be made on the basis of imperfect retrospective data. Recommendations for surgical strategy in differentiated thyroid cancer are discussed. PMID- 9221636 TI - [Significance of lymph node metastases of differentiated thyroid gland carcinomas and C-cell carcinomas for prognosis--a meta-analysis]. AB - Surgical therapy of differentiated thyroid cancer (DTC) includes thyroidectomy plus central lymph node dissection and postoperative radioiodine therapy. In cases of lymph node metastasis, T3/T4 tumors and C-cell-carcinoma (after thyroidectomy) uni- or bilateral modified radical lymph node dissection of the neck (neck dissection) and of the mediastinum is recommended. The importance of lymph node metastasis for prognosis of survival in papillary, follicular and C cell-carcinoma is discussed controversial, however. Even the kind of surgical radicality is questioned. Thus a metaanalysis of 35 studies in 29 independent publications from a pool of 2186 studies was performed. Univariate analysis demonstrates lymph node metastasis as a negative prognostic factor in papillary carcinoma with a 3.25/2.97, in follicular carcinoma with a 7.62/4.0 and in C-cell carcinoma with a 3.33/3.37 higher probability of mortality 5 and 10 years after operation. Modification of the present surgical therapy can therefore only be accepted after univariate and multivariate analysis of all prognostic factors (age, sex, cell type, tumor extent, lymph node- and distant metastasis) and after it has proven superiority to the present strategy in prospective randomised trials. PMID- 9221637 TI - [Radioiodine therapy in differentiated thyroid gland carcinoma]. AB - The current level of knowledge about radioiodine therapy (RITh) for well differentiated thyroid carcinoma under consideration of the recent literature is summarised. In RITh for thyroid carcinoma two major fields can be distinguished: the ablation of the thyroid remnant and the therapy of recurrences resp. local and distant metastases. New large American studies indicate, that the prophylactic post operative ablation of the thyroid remnant in primary tumours over 1.5 cm in diameter is linked with a distinct improvement of the long-term prognosis. The RITh is effective also in distant metastases, if applied early, when the tumour volume is still small. The prerequisite is an appropriate follow up. Surveys about the application of RITh for well-differentiated thyroid carcinoma in Europe and in the USA reveal that uniform treatment recommendations designed to conform interdisciplinary demands are urgently required. PMID- 9221638 TI - [Radioiodine therapy and radioiodine after-care in differentiated thyroid gland carcinomas]. AB - Differentiated thyroid carcinoma (DTC) is a rare tumor with a generally good prognosis. Still some of these cancers cause the patient's death after many years of illness. Thus, treatment and aftercare have to be risk-adapted according to tumor type and staging. Surgery is the primary treatment for this tumor and its metastases, followed by therapy with radioiodine. According to the guidelines edited by the German Association for Nuclear Medicine (DGN)--work group for therapy--radioiodine therapy is an effective regimen with minimal side-effects. Radioiodine scintigraphy is the most important part of the aftercare program, together with the monitoring of the highly specific tumormarker thyreoglobulin. Other diagnostic modalities may be added when considering therapeutic options or confirming clinical suspicions. This paper aims at giving an update and overview on the applications of radioiodine, other radioisotopes and additional diagnostic and therapeutic options in differentiated thyroid cancer and related aspects of radiation exposure and radiation protection. As new therapeutic procedures and optimised diagnostic modalities become available, there is new hope for patients with locally advanced or metastatic DTC. PMID- 9221639 TI - [Postoperative benefit after laparoscopic cholecystectomy in acute cholecystitis]. AB - Analysed were the results of all 48 patients with acute cholecystitis, who underwent laparoscopic cholecystectomy between 1991 and 1995 in the department of general surgery, AKH, University of Vienna. In 18 cases it was necessary to convert to laparotomy. In a second step the results of these two groups of patients were compared with results of patients without acute cholecystitis, who elective underwent laparoscopic cholecystectomy. In about 2/3 of the patients with acute cholecystitis laparoscopic cholecystectomy is possible. In these cases we found a mean postoperative hospital stay of 4.4 days, with a significant difference between those with drain (5.9 days) and those without (2.7 days). In cases of laparotomy the mean postoperative stay was 7.7 days, also significant longer. These patients consumed at the first postoperative day more than 1.5 times of opioid analgetics than those, who underwent laparoscopy with acute cholecystitis, laparoscopied patients without acute cholecystitis half of this. After the second postoperative day patients after laparotomy took 3 times of opioid analgetics than patients after laparoscopy, no matter if there was an acute cholecystitis or not. The rate of conversion to laparotomy sank with the increase of experience of a surgeon. Postoperative benefit of laparoscopic treatment, as less pain and shorter hospitalisation, can be saved even in cases of acute cholecystitis. So the management should primary be laparoscopic. PMID- 9221640 TI - [Laparoscopic splenic surgery--report of 17 personal cases]. AB - Since the beginning of the decade we can remark an increasing of publications about laparoscopic approach in splenic surgery. We report about 17 unselected own cases (8 males, 9 females). Main indications were 12 hematologic diseases: 9 idiopathic thrombopenic purpuras, 1 hemolytic anemia, 1 Hodgkin's disease and 1 recurrent splenic infarction with hypersplenism and extramedulary hemapoiesis, further 1 unclear sepsis with focal lesions and splenomegaly, 1 dysontogenetic splenic cyst, 1 splenic co-affection of Wegener's disease, and 2 traumatic lesions. We performed in 13 cases asplenectomy, under these 1 time combined with a laparoscopic hernioplasty and 1 time with a complete staging, further in 1 case a partial splenic resection, in 1 case a diagnostic excision and in 2 cases a hemostyptic management without splenectomy. We had to convert in no case, the lethality rate was also 0. As a complication we noted one postoperative bleeding, we could managing also laparoscopically. Our mean operating time of 137 min was evident shorter than the published ones by other authors. Because of our good results and the excellent tolerance by patients we can recommend this proceeding. Nevertheless, it should be limited for surgeons with a large experience at laparoscopic centers. PMID- 9221641 TI - [Extraction of cystic duct occlusion calculus in laparoscopic cholecystectomy]. AB - Occluding stones left in the stumpf of the cystic duct may account for between 17% and 25% of the cases of post-cholecystectomy syndrome. When acutely inflamed or empyemic gallbladders are removed, an occlusive cystic duct stone must almost always be expected. When performing laparoscopic cholecystectomy, therefore, care must be taken to ensure that any stone occluding the cystic duct are detected and removed. After completely freeing the cystic duct to the point of its junction with the common bile duct, it is carefully "palpated" with a 5 mm forceps for the presence of stones. Any such present are pressed out of the duct through a transverse incision, and retrieved. Bile reflux through the incision in the duct indicates freedom from stones. Finally, intraoperative cholangiography can be performed. PMID- 9221642 TI - [Spontaneous intraperitoneal perforation of the urinary bladder--a rare cause of acute abdomen. 2 case reports]. AB - In the last ten years we have seen two cases from which we can prove that the cause of the acute abdomen was an intraperitoneal urinary bladder perforation. One had previously undergone radiation treatment of the abdomen and the other had a total abdominal hysterectomy. PMID- 9221643 TI - [Modification of energy supply by pancreatic mitochondria in acute experimental pancreatitis]. AB - A disturbed energy metabolism in pancreatic acinar cells is discussed as factor contributing to the development of acute pancreatitis (AP). In this study, we investigated to what extent the mitochondrial ATP producing capacity is impaired in the pancreatic tissue of rats with experimental AP. For preparation of mitochondria from rat pancreas, routine isolation procedures (tissue homogenization and differential centrifugation) were applied. Mitochondria were isolated from rats with edematous pancreatitis produced by hyperstimulation with caerulein, and from rats with mild necrotizing acute pancreatitis. The latter form of AP was induced by a temporary occlusion of the biliary pancreatic duct accompanied by a simultaneous intravenous injection of caerulein plus secretin and an intraabdominal administration of ethanol. As functional parameters of oxidative phosphorylation, the respiration rate, the mitochondrial membrane potential, and the activity of the complex I of the respiratory chain were determined. Mitochondria from rats with caerulein AP showed an enhanced respiration (61% vs. saline control) and a diminished membrane potential (-17 mV) if respiring with succinate in the non-phosphorylating state. This indicates an increased proton leak across the mitochondrial inner membrane. In the mild necrotizing AP, mitochondria were characterized by a decreased respiration with NAD(+)-linked substrates (-33% vs. sham-operated animals). This inhibition of respiration was confirmed by the reduced activity measured for the NADH cytochrome c reductase (-32%). In both models of experimental AP the potency of mitochondria to produce ATP was significantly diminished. The stronger impairment of mitochondrial functions were found in the necrotizing form of AP. Reactive oxygen species may lead to the observed alterations--to the enhanced permeability of the mitochondrial inner membrane as well as to the inhibition of the complex I of the respiratory chain. PMID- 9221644 TI - [Response to the letter by Dr.-H. Zermann on the contribution by W.H. Jost, "Value of electromyography in diagnosis of incontinence"]. PMID- 9221645 TI - [The mechanism of the existence of Yersinia pestis in nature]. PMID- 9221646 TI - [Microbiology and Nobel Prize winners]. PMID- 9221647 TI - [An outbreak of acute intestinal infections in a nursery-kindergarten (2)]. PMID- 9221648 TI - [The role of the outer-membrane antigens of Yersinia pseudotuberculosis in the pathogenesis and diagnosis of pseudotuberculosis]. AB - The degree of manifestation of the virulent properties of Y. pseudotuberculosis strains was found to be directly related to the concentration of protein with a mol. wt. of 103 kD (invasin), localized in the outer membrane and necessary for the realization of the process of the penetration of the infective agent into eukaryotic cells. Antiserum to INV-2-BSA, an active synthetic fragment of invasin, capable of detecting one antigenic band with a mol. wt. of 103 kD, was used for the creation of a diagnostic enzyme immunoassay system. The new variant of the diagnostic system was shown to be highly effective in the diagnosis of the disease at an early period. PMID- 9221649 TI - [The use of high-capacity immunoplates in determining antibodies to the human immunodeficiency virus]. AB - The action of gas plasma on the sorption characteristics of polystyrene immunoplates is described. The treatment of immunoplates with gas plasma has proved to be a highly effective method for increasing their adsorption capacity and the sensitivity of immunoassay in the determination of antibodies to human immunodeficiency virus (HIV-1). This method makes it possible to increase the sensitivity of solid-phase immunoassay for the determination of a number of viral infections. PMID- 9221650 TI - [An outbreak of nosocomial candidiasis in a hematology ward]. AB - The aim of the present investigation was the study of the epidemic process of hospital candidiasis. The study was carried out from October 1991 to May 1992 in a hematological ward of one of the hospitals of St.-Petersburg on 9 patients with generalized forms of candidiasis: in 4 of them the disease was caused by C. albicans and 5--by C. parapsilosis. Intestinal dysfunction as one of the first manifestations of candidosepsis caused by C. albicans, the isolation of C. albicans from feces, ulcero-necrotic colitis (established by postmortem examination), as well as the incapacity of these patients of immune response (in 2 serologically examined patients the titer of antibodies to fungi of the genus Candida was found to be lower than the diagnostic titer), were indicative of the dissemination of fungi from the gastrointestinal tract. The cases of fungemia caused by C. parapsilosis were due to the fungal contamination of vascular catheters in these patients. This infective agent was also isolated from washings obtained from the hands of nurses carrying out treatment and from medical rubber gloves. Moreover, some violations in the rules of the preparation of disinfecting solutions and the treatment of the hands of the personnel were established. PMID- 9221651 TI - [The possibilities for using combined methods in typing Candida albicans for establishing epidemiological relationships]. AB - An outbreak of candidiasis caused by C. albicans was registered in a neonatal intensive care unit of one of the hospital of St.-Petersburg at the period of August 12-October 8, 1992. C. albicans were isolated from 21 out of 87 patients. To achieve their more detailed deciphering and to establish epidemiological relationships, the intraspecific typing of C. albicans was carried out by DNA typing and by the electrophoretic spectrum of exoproduct proteins. The analysis of the results of this typing revealed that strains classified with the same biotype belonged to different electrophoretic groups and, vice versa, strains of the same electrophoretic group could belong to different biotypes. The analysis of the data obtained by taking into account the results of typing by the two methods, the time of the patients stay in the unit, the number of the maternity clinic from which an individual patient arrived, the number of the reanimation place and the number of the apparatus for artificial lung ventilation made it possible to establish epidemiological relationships between the patients in all cases. PMID- 9221652 TI - [The role of the thrombocytic cationic protein (beta-lysin) in anti-infectious protection]. AB - The biological activity of beta-lysin, cationite protein of thrombocytic origin with bactericidal activity, isolated from human blood serum was studied. The mechanisms of its bactericidal action was determined: the inhibition of bacterial catalase and peroxidase. The study revealed that the activity of beta-lysin in anti-infectious protection was due to both its direct bactericidal action and the enhancement the phagocytic reaction of the body by the stimulation of phagocytes and the opsonization of the pathogen. PMID- 9221653 TI - [Shigellosis in Moscow: new trends in the epidemiological process and their social determinants]. AB - In Moscow for a long time morbidity in acute enteric infections, and in Shigella infections as their main constituent, steadily decreased. In recent years the opposite tendency was noted: a rise in morbidity. A pronounced tendency for Shigella infections, especially Flexner's dysentery, to affect older ages was observed. New risk groups, including adults busy in "shuttle" commerce and different asocial elements, appeared. The number of lethal cases considerably increased. Unusual manifestations of the epidemic process are socially determined and reflect a decrease in the living standards of the population, sharp social changes in its way of life and social instability in the country. Sporadic morbidity with low focality prevails. The so-called chronic multifactor alimentary route of infection transmission is mainly realized; its neutralization presents great objective difficulties. PMID- 9221654 TI - [The results of monitoring enterovirus circulation circulation among the population and in the environment of Tula Province over 10 years (1985-1994)]. AB - The study of the isolation rate of polioviruses and other enteroviruses in patients with different diagnosed diseases, among healthy child population, as well as the circulation of these viruses in the environment was carried out on the territory of Tula Province for the period of 1985-1994. The epidemiological analysis of the data obtained in this study are presented. The study revealed that the vaccinal prophylaxis of poliomyelitis, carried out for the period of many years, did not lead to the elimination of poliovirus strains, differing in their genetic properties from vaccine strains, on the territory of Tula Province. A decrease in the immune stratum with respect to polioviruses of types I, II and III, observed in 1985-1994, was accompanied by the circulation of polioviruses of these three types among the population. PMID- 9221655 TI - [The isolation amd study of lipopolysaccharide from Klebsiella pneumoniae vaccinal strain 204]. AB - K. pneumoniae lipopolysaccharide (LPS) was isolated from biomass grown under the conditions of controlled multicycle and continuous cultivation. A considerable yield of LPS containing the minimal amount of protein and nucleic acid admixtures was obtained from biomass grown by continuous cultivation with the concentration of glucose in the medium equal to 20 g/l and the content of dissolved oxygen at a level of 0% of complete saturation. Erythrocyte diagnosticum prepared on the basis of this LPS was found to have a sensitizing dose of 100 micrograms per ml of solid erythrocytic precipitate and high specificity, studied in the passive hemagglutination test with the use of rabbit sera. PMID- 9221656 TI - [The effect of the processes of Klebsiella pneumoniae cultivation on the antigenic activity of vaccinal preparations for peroral use]. AB - Whole-cell preparations, obtained from the microbial mass of K. pneumoniae grown by batch and continuous cultivation at a dilution rate of 0.24 and 0.41 hr-1 and used in animal experiments for oral administration, were shown to have different serological activity. The preparation obtained from biomass grown by continuous cultivation at a dilution rate of 0.41 hr-1 proved to be most active regarding the level of hemagglutinating antibodies to K. pneumoniae LPS. At the same time the 360-fold rise of the level of anti-LPS antibodies in rabbit immune sera was observed. On day 258 oral revaccination was made; after that the twofold rise of the level of anti-LPS antibodies in the sera of the animals was observed. These antibody levels exceeded 40-fold those registered before primary immunization, and sufficiently high antibody levels were retained for 4 more months (the term of observation). PMID- 9221657 TI - [The dynamics of the formation of IgG subclasses in Balb/c and CBA mice infected with the LCM, Lassa and Mopeia viruses and their role in diagnosis]. AB - The dynamics of the induction of individual IgG subclasses in BALB/c and CBA mice and their role on the diagnostics of arenaviruses LCM, Lassa and Mopeia was studied. The study demonstrated that in solid-phase enzyme immunoassay (EIA) isotypes IgG2a and IgG2b to virus Mopeia could be used for the differentiation of virus Mopeia from viruses LCM and Lassa, and the antigen of virus Mopeia could be used for the preparation of diagnostic EIA systems not only to nonpathogenic virus Mopeia, but also to pathogenic viruses LCM and Lassa. PMID- 9221658 TI - [Methods for assessing the efficacy of immunocorrection]. AB - The use immunocorrective remedies has become a topical problem due to the insufficient effectiveness of the treatment of many chronic diseases (including chronic purulent otitis), manifested by the suppression of initial immunological disturbances. Still the principles of the prescription of the above-mentioned remedies have not yet been sufficiently worked out. The present work deals with the use and approval of the methods for the evaluation of the effectiveness of immunocorrective therapy, the determination of the targets of action, the proper effect of modulation, changes in the degree of its manifestation. PMID- 9221659 TI - [The comparative characteristics of the effect of immunomodulators on the indices of body nonspecific protection and on the structure of the populations of the causative agents in mixed infections]. AB - A sharp decrease in the characteristics of nonspecific protection and humoral immunity in mixed infections (Pseudomonas aeruginosa + fungi of the genus Candida) in cases of burn traumas was established. Under these conditions an increase (up to 100%) in the number clones with the signs of pathogenicity and multiple medicinal resistance in the populations of infective agents, accompanied by a high death rate among the animals (up to 60%), was observed in the dynamics of the infectious process. The use of immunomodulators (tactivin and sodium nucleinate + lidocaine) contributed to the restoration of the phagocytic function of peripheral blood lymphocytes and humoral immunity, as well as to a decrease in the death rate of the animals. At the same time a considerable decrease in the number of clones with the signs of pathogenicity was observed in the populations in infective agents, isolated from the animals in the process of treatment. PMID- 9221660 TI - [The spread of antibodies to cytomegalovirus and Toxoplasma among donors of blood components]. AB - The spread of IgG antibodies to cytomegalovirus and Toxoplasma in the blood sera of 323 donors of hemocomponents was studied. Among young males in the northwestern region of Russia three fourths were found to be seropositive to cytomegalovirus and one third, to Toxoplasma. In older ages the proportion of seropositive persons somewhat increased. In donors with blood group O (I) antibodies to cytomegalovirus occurred almost twice as often (91%) as in donors with blood group AB (IV) (47%). Among donors with blood group AB (IV) the proportion of persons seropositive to Toxoplasma (54%) was twice as great as that among donors with blood group O (I) (27%). PMID- 9221661 TI - [The physicochemical characteristics of ribosomal fractions from Pseudomonas aeruginosa obtained by precipitation with polyethylene glycol 6000]. AB - Ribosomal fractions containing up to 72% of ribosomal material and 25% of sugars (among them, about 6% of hexose) were isolated from P.aeruginosa cells, immunotypes F-1, 2, 6 and 7, by precipitation with polyethylene glycol 6000. Lipopolysaccharide, determined in the test for ketodesoxyoctanoic acid, was not detected in these fractions, but, as determined in the passive hemagglutination test, the content of O-antigen in the preparations was 3-25%. O-antigen and ribosome present in the fractions formed a complex, disintegrating after treatment with trilon B. PMID- 9221662 TI - [The functional metabolic activity of the blood neutrophils in different clinical forms of brucellosis]. AB - The characteristics of the activity of alkaline phosphatase, nonspecific esterase, myeloperoxidase, lysosomal cation proteins, lipids have been determined in the cytochemical study of neutrophils in peripheral blood samples of 127 patients with acute, chronic, primary chronic brucellosis and osteochondrosis. It is expedient to use these characteristics as additional tests for the differential diagnostics of different clinical forms of brucellosis, as well as residual brucellosis and osteochondrosis. PMID- 9221663 TI - [The identification of salmonellae by means of a rapid polymerase chain reaction]. AB - The results of the rapid method for the detection and identification of epidemically related and unrelated causative agents of Salmonella infections. To analyze the chromosomal DNA of these strains, rapid polymerase chain reaction (PCR) was used. The analysis of the electrophoregrams of products obtained in rapid PCR revealed that the number of DNA fragments is constant for the cultures of the concrete serovar or phagovar. Using rapid PCR, the detection of S. typhi atypical strains was succeeded. PMID- 9221664 TI - [The possibility of colonizing the intestines of white mice with Lactobacillus acidophilus during bacterial therapy]. AB - The study revealed the possibility, on principle, for L. acidophilus strain VKM V 2020 D to colonize the intestine of white mice with the preservation of the viability of lactobacilli subjected to the action of antibiotics. The culture of this strain, isolated from the animals, showed the stability of its biological properties: resistance to polymyxin M, kanamycin, cyprofloxacin, nalidixic acid (including acquired resistance to rifampicin), as well as pronounced antagonism with respect to Vibrio cholerae. Good prospects for the use of L. acidophilus strain VKM V-2020 D for further studies regarding its use for prophylaxis and therapy were noted. PMID- 9221665 TI - [Viral hepatitides from A to G and further]. PMID- 9221666 TI - [Polynucleotide (DNA) viral vaccines and their safety problems]. PMID- 9221667 TI - [The isolation of the surface antigen from vegetative cells of Bacillus anthracis STI-1 and study of its protective properties]. AB - The method for the extraction of native surface protein antigen with a mol. wt. of 92 kD from vegetative cells of B.anthracis STI-1 and its purification was developed. The antigen was extracted with 3% sodium lauryl sarcosylate at 4 degrees C from bacterial mass previously treated with 0.1 M tris-HCl buffer solution, pH 8.0 Purification was carried out by adsorption chromatography on hydroxylapatite. The isolated protein antigen with a mol. wt. of 92 kD was electrophoretically and immunochemically homogeneous. The study of the protective properties of this protein in combination with Freund's adjuvant, made on white mice, revealed that it had good prospects for use in the creation of chemical vaccines against the causative agent of anthrax. PMID- 9221668 TI - [The role of plasmid gef-like genes in eliminating from bacterial populations the specimens that have lost these plasmids]. PMID- 9221669 TI - [The definition and subject of modern epidemiology]. PMID- 9221670 TI - [The causative agents of chlamydiosis in agricultural animals and their pathogenicity for man]. PMID- 9221672 TI - A gift disguised. PMID- 9221673 TI - Telehealth discussions focus on licensure. PMID- 9221671 TI - Unexpected alcohol withdrawal. PMID- 9221674 TI - Nursing's role in the assisted suicide debate. PMID- 9221675 TI - Abstracts of the combined scientific meeting of the Australian and New Zealand College of Anaesthetists, Faculty of Intensive Care and Australian Society of Anesthetists. Western Australia, October 26-30, 1996. PMID- 9221676 TI - [Possibilities of transcutaneous monitoring of blood gases]. AB - The authors analyze the experience gained in noninvasive transcutaneous monitoring of blood gases ptkO2 and ptkCO2 (MICROGAS device, Contron, France) in 33 oncological patients during surgery with irradiation of the operation wound (n = 13) under conditions of total intravenous anesthesia (TIA) with artificial ventilation of the lungs with a hypoxic respiration mixture (FiO2 10%) and during surgery for breast cancer under TIA with spontaneous respiration (n = 20). Transcutaneous monitoring was found to be a sensitive rapidly reacting method, more informative than pulsed oximetry, for intraoperative hypoxyradiotherapy with long-distance monitoring of a patient; under such conditions it can be the only possible method for monitoring the preset level of pO2. The results of measuring ptkO2 and ptkCO2 are close to the values in the arterial blood. PMID- 9221677 TI - [Gas dynamics characteristics of current methods of jet ventilation of the lungs]. PMID- 9221678 TI - [Mechanisms of acute respiratory insufficiency in patients with hyperglycemic coma]. AB - Basic parameters of pulmonary gas exchange, central and pulmonary hemodynamics, and colloid osmotic pressure were investigated in 31 patients in diabetic hyperglycemic coma over the course of intensive care. Pulmonary gas exchange disorders were observed in all patients in the presence of increased shunting of the blood in the lungs and disorders of transcapillary liquid exchange. On the other hand, we failed to obtain data indicative of an increase in the volume of extravascular water in the lungs. However, it does not rule out the possibility of iatrogenic disorders of gas exchange during noncontrolled rehydration. PMID- 9221679 TI - [Changes in central hemodynamics and their correction during reconstructive surgery in Leriche's syndrome]. AB - Study of the parameters of central hemodynamics in the course of aortofemoral shunting for Leriche's syndrome revealed the role of lipid peroxidation dysfunction in the genesis of cardiovascular disorders developing in the immediate postoperative period. The postoperative period is characterized by expressed hypotension developing in the presence of activation of lipid peroxidation and suppression of plasma antioxidative activity. The authors recommend alpha-tocopherol, cytochrome C, and dalargin to prevent these hemodynamic disorders. PMID- 9221681 TI - [Stunned myocardium: an experimental phenomenon or a clinical syndrome?]. PMID- 9221680 TI - [Components of anesthesiological provision and central hemodynamics in hypothermia without perfusion]. AB - Central hemodynamics was studied by thermodilution in 16 patients with acquired mitral defect during surgery under conditions of hypothermia without perfusion. Cooling and surgical correction were carried out under superficial ether anesthesia with morphine and droperidol. A 28% decrease of the minute circulation volume is observed as early as under general anesthesia and at the very beginning of cooling (10.8% more); this decrease is caused by rarer heart contractions and increase of the total peripheral vascular resistance as a result of body response to cool exposure. An almost 10 degrees C drop of body temperature (to 27.2 +/- 0.2 degrees C) involves a just 13% decrease of the cardiac index in comparison with the value at the beginning of cooling. Injection of droperidol before cooling blocks the increase of the total peripheral resistance in a dose dependent mode (about 0.13 mg/kg of droperidol is needed for sufficiently complete prevention of the increase of total peripheral resistance), and thus maintains the stroke and cardiac indexes at a sufficiently high level. Morphine (in doses of up to 2.3 mg/kg) did not bring about such an effect. PMID- 9221682 TI - [Assessment of microcirculation using mathematical models of hydrodynamics and oxygen transport in microvessels. I]. PMID- 9221683 TI - [Hyperbaric oxygenation and antiaggregants: effects on platelet function in patients with ischemic heart disease]. AB - Platelet function (duration and percentage of aggregation and disaggregation) was studied in 65 chronic coronary patients over the course of hyperbaric oxygenation and antiaggregant therapy. A course of hyperbaric oxygenation (8 to 12 sessions at absolute atmosphere of 1.3-1.6 for 40 min) had no proaggregant or antiaggregant effects on the platelets in this category of patients. Aspirin in a daily dose of 125 mg and pentoxifylline in a daily dose of 300 mg reliably decreased the percentage of platelet aggregation and increased the share of disaggregation. Hyperbaric oxygenation in the above mode did not change the antiaggregant activity of these drugs. PMID- 9221684 TI - [State of the hypophyseal-adrenal system during surgical correction of heart defects under various anesthesiological methods and at distant periods after surgery]. AB - The concentration of 11-HOCS in the blood plasma was measured in 278 patients with heart diseases operated on under various anesthesias at different stages of surgical correction of the defects and in the immediate postoperative period. Plasma ACTH level was measured in 137 of these operated on under deep hypothermia and hypothermal perfusion. The hormonal response was the most expressed in patients operated on under normothermia, whereas in patients exposed to hypothermia the reaction was minimal. 141 patients were examined in remote periods after surgery. Blood plasma 11-HOCS level was normal in the cases with good and satisfactory results of surgical treatment, whereas in those with poor results it was higher or lower than normally. PMID- 9221685 TI - [Effects of mental adaptation on vegetative, hormonal and sensory reactions in preoperative period]. AB - Fifty-seven urological patients whose emotional behavioral reactions were characterized by somatic anxiety caused by mental adaptation were examined under conditions of preoperative emotional stress. On day 5 after admission to hospital the hormonal, autonomic, and sensory responses decreased in patients with previously manifest anxiety, which was due to decrease of anxiety because of appearance of some psychopathological features. Satisfactory hormonal, autonomic, and mental adaptation was observed on day 5 after admission to hospital in patients with moderately expressed anxiety. In patients with previously predominating parasympatotonia and a low level of anxiety, the hormonal and autonomic areactivity and mental adaptation observed in the same terms were due to augmenting psychopathological features. PMID- 9221686 TI - [Criteria in discontinuing artificial ventilation of the lungs after planned and emergency surgical interventions]. PMID- 9221687 TI - [Brain metabolism in patients undergoing surgery under conditions of artificial blood circulation]. PMID- 9221688 TI - [Changes in bioelectric activity and metabolism of the brain during surgery of the aorta under deep hypothermic arrest of circulation]. AB - Bioelectric activity and metabolism of the brain were studied during surgery on the aorta with deep hypothermal arrest of circulation. The study included 9 patients (7 men and 2 women) aged 13 to 66 years. The mean duration of circulation arrest under deep hypothermia was 48.6 (from 19 to 77) min. Before circulation arrest the patients were cooled to nasopharyngeal temperature of 13.5 +/- 0.5 degrees C and rectal temperature of 15 +/- 0.6 degrees C. Deep hypothermia involved the disappearance of bioelectrical activity and decrease of the spectral power of all EEG frequency bands. The most notable changes in the hemoglobin saturation of the blood in the internal jugular vein bulb (SibO2) were observed during artificial circulation and hypothermal arrest of circulation. Cooling of patients led to a gradual increase of its values. Before deep hypothermal arrest of circulation SjbO2 was as high as 98.4 +/- 0.4%. Over the period of circulation arrest its level dropped to 85.7 +/- 4.8%. Subsequent warming led to its further decrease, and by the end of artificial circulation it was as initially. The content of carbon dioxide in the blood flowing from the brain increased from 17.0 +/- 0.9 to 31.7 +/- 4.7 mm Hg over the period of heart arrest, which may be indicative of the continuing metabolic processes in the brain under conditions of deep hypothermia and justifies the additional drug protection and local hypothermia of the head. PMID- 9221689 TI - [Catheterization of the internal jugular vein in the assessment of brain metabolism: right or left side?]. AB - Oxygen and glucose were measured in arterial and venous blood of 10 patients operated on for arterial aneurysms of the cerebral vessels. The blood was collected through catheters inserted on the right and left from the internal jugular vein bulb. Blood samples were collected at the beginning and end of surgery simultaneously from 3 sites: artery and right and left veins. The values for the right and left jugular veins initially differed much in 5 out of 11 cases. The causes of these differences are discussed, but this fact makes us doubt the efficacy of catheterization of one internal jugular vein for assessing the cerebral metabolism in various clinical situations. PMID- 9221690 TI - [State of prostanoid-regulating function of the lungs in patients subjected to thoracic surgery]. AB - Lung capacity to regulate the level of plasma prostanoids at the expense of their destruction or extra synthesis is one of their numerous non-gas-exchange functions. Prostaglandins A, E, and F, prostacyclin, and thromboxane were measured in the arterial and venous blood of 23 patients before and after surgery. The level of prostanoids was sharply increased in surgical patients. Substrates with the broncho- and vasoconstrictive action predominate in the blood of patients with obstructive and restrictive changes in the lungs, this eventually leading to complications in the postoperative period. PMID- 9221692 TI - [Effects of the initial composition of perfusate on lactate values during extracorporeal circulation]. PMID- 9221691 TI - [Fatal acute hemorrhage and activity of serum and erythrocyte cholinesterase]. PMID- 9221693 TI - [Preoperative efferent therapy of patients with atherosclerosis obliterans of the lower limbs]. PMID- 9221694 TI - [Monitoring neuromuscular conduction by the accelerometric method in anesthesiology]. PMID- 9221695 TI - [Safety monitoring. 5-year experience of use in anesthesiology and intensive care]. PMID- 9221696 TI - [Anesthesiological practice in Scotland]. PMID- 9221697 TI - Proceedings of the 18th Symposium on Biotechnology for Fuels and Chemicals. Gatlinburg, Tennessee, May 5-9, 1996. PMID- 9221698 TI - Towards Minimally Invasive Coronary Artery Bypass Grafting. Proceedings of the 2nd Utrecht MICABG Workshop. Utrecht, the Netherlands, October 4-5, 1996. PMID- 9221699 TI - [A comparative study of fluoroquinolones and 3rd-generation cephalosporins in the prevention and treatment of experimental plague caused by Yersinia pestis strains typical and serologically atypical with respect to F1]. AB - Fluoroquinolones (ciprofloxacin and pefloxacin) and 3rd generation cephalosporins (cefoperazone, cefotaxime, ceftazidime and ceftriaxone) were comparatively studied in the prevention and treatment of experimental plague in albino mice caused by F1+ and F1- strains of the plague microbe. Despite the phenotype of the strain which caused the infection, the drugs were highly efficient in the etiotropic therapy. However, in the experimental plague due to F1- strains it was needed to use the maximum mean daily doses of the fluoroquinolones, cefoperazone and cefotaxime. For the prevention of the infection such doses should be used for not less than 7 days. By the efficacy ceftriaxon was superior to the other cephalosporins and should be considered as a drug of choice. PMID- 9221700 TI - [The effect of calcium gluconate on the acute and chronic toxicity of doxorubicin in mice]. AB - The effect of calcium gluconate on the toxicity of the anthracycline antibiotic doxorubicin (DOX) in mice was studied. Calcium gluconate showed a significant protective action with respect to the DOX acute toxicity. When DOX was used in the lethal doses, up to the LD50s calcium gluconate protected all the mice from death. At the DOX LD100 or higher the antitoxic effect of calcium gluconate manifested itself in a lower death rate and/or in a higher lifespan of the animals (at least 2-fold). When DOX was used for the treatment course its chronic toxicity in the presence of calcium gluconate was 2 or more times lower by all the quantitative indices: the lifespan of the animals that died, the maximum and minimum total lethal doses of DOX, the latent period before the first mouse death, the overall duration of the DOX treatment course. The antitoxic effect of calcium gluconate also manifested itself in a lower DOX acute and chronic toxicity with respect to the gastrointestinal tract. Thus, calcium gluconate proved to be an effective DOX antitoxic modificator which provided the use of about 2 times higher single and courses doses of DOX in mice. PMID- 9221701 TI - [The pharmacokinetics of fluoroquinolones in different diseases]. AB - A literature review on the pharmacokinetic characteristics of fluoroquinolones in the treatment of patients with various diseases such as diseases accompanied by hepatic insufficiency, mucoviscidosis, diseases in elderly patients, lower respiratory tract infection, septicemia, skin infection and others was analyzed. Infections requiring correction of the routine treatment regiments are indicated. PMID- 9221702 TI - [Our journal is 40 years old. Antibiotiki i Khimioterapiia]. PMID- 9221703 TI - [The treatment of pneumonia: the choice of the initial preparation]. PMID- 9221704 TI - [Pasteurella multocida: infections in man]. AB - The literature data on human infection due to a representative of the genus Pasteurella, i.e. P. multocida are reviewed: the main clinical forms and signs of pasteurellosis, the results of the studies on susceptibility of P. multocida to antibiotics and chemotherapeutics and the results of their use in the treatment of the infection. PMID- 9221705 TI - [The isolation and characteristics of mutants of the Saccharopolyspora erythraea strain resistant to thiostrepton]. AB - The formation of thiostreptone resistant spontaneous and nitrosoguanidine-induced mutants in the erythromycin-producing organism Saccharopolyspora erythraea was investigated. The investigated collection of the mutants was heterogeneous by the level of the thiostreptone resistance (2.5 to 20 micrograms/ml). The thiostreptone resistance mutations had a pleiotropic effect: 17 per cent of the mutants was characterized by the growth thermosensitivity and 26 and 5.8 per cent of the mutants were characterized by loss of the ability to form melanine and aerial mycelium respectively. Such phenotypes were most frequent in the mutants resistant to low concentrations of thiostreptone (2 to 5 micrograms/ml). The absolute majority of the isolated thiostreptone resistant mutants was unstable and formed both the antibiotic resistant and the antibiotic sensitive clones. The greatest portion of the strains with high antibiotic activity (20 per cent) was detected among the S. erythraea spontaneous mutants on the medium with 2.5 micrograms/ ml of thiostreptone. It was shown that the instability of the high antibiotic activity in the mutants was associated with loss of the thiostreptone resistance property. PMID- 9221706 TI - [The relationship between Chlamydia pneumoniae and atherosclerotic lesions of the aorta]. AB - Although atherogenic main factors have been extensively studied, there are others whose real importance has not been well defined. There are some pathologic and immunologic evidences relating several infectious agents with the genesis or development of coronary atherosclerosis. Recently, a link has been established between Chlamydia pneumoniae and atherogenesis, due to immunological evidence of infection in human atherosclerotic lesions. We studied 16 aortic specimens obtained from necropsies performed in subjects who died with coronary heart disease. The infection of Ch. pneumoniae was determined by means of an immunofluorescent technique using a specific monoclonal murine antibody. A positive reaction was found in advanced non-ulcerated fibrolipid lesions in just 2 patients (13%), according with several other observations. It is not known the true relationship between the chlamydia infection and atherogenesis, neither if the infection starts or aggravates the atherosclerotic process or if it is an independent phenomenon. PMID- 9221707 TI - [Coronary stents. Results during the hospitalization phase]. AB - Between October 1991 and August 1996 two-hundred coronary stents were implanted (s) in 166 patients (pts) (1.27 s/pt). One hundred thirty-five lesions "de novo" were approached with stent, 44 because of a sub-optimal result post-angioplasty, 15 for restenosis and 17 for dissection. In thirty-six patients the indication of stenting was stable angina, in 68 unstable angina, in 37 for angina after myocardial infarction, in 11 for asymptomatic ischemia after myocardial infarction and in 14 during an acute myocardial infarction. Mean stenosis before stent implantation in all cases was 85 +/- 15%. Type of lesion in seventy cases was A, in 112 was B and 29 was C. Stents used were AVE in 146 lesions, Palmaz Schatz in 33, Wiktor in 23, Gianturco-Roubin in 8 and Wallstent in one case. Medical treatment in 140 pts. (84.3%) after stent implantation was only with aspirin and ticlopidine. Technical success in all patients was 98.6% (208/211 pts) and primary success was 94.6% (157-166 pts). Unsuccessful procedures were because of sub-acute occlusion in three patients (1.8%), death in 3 pts. (1.8%) and urgent CABG was necessary in one pt. (0.5%). Major hematoma was a complication in 5 pts (3%). Mean residual stenosis after stent implantation in all cases was 2.2%. CONCLUSION: Stent implantation in our laboratory is a very safe procedure with a high rate of primary success with lowest complications in a great population of unstable angina. PMID- 9221708 TI - [Ambulatory monitoring of arterial pressure in the treatment of mild-moderate arterial hypertension with lisinopril vs. enalapril]. AB - OBJECTIVE: To compare the antihypertensive efficacy of lisinopril (L) versus enalapril (E) given 20 mg once daily for the treatment of mild to moderate arterial hypertension (diastolic blood pressure between 90-114 mm Hg) using ambulatory blood pressure monitoring (ABPM). This study also investigated the tolerance and changes in haematological or biochemical parameters with both drugs. METHODS: One hundred patients (men and women) with a range of age between 18 to 75 years were included in this open, randomised study to assess the hypotensive efficacy of L versus E after 2 months of treatment, using ambulatory blood pressure monitoring. To study data obtained from ABPM Mc Call curves were used and statistical analysis was made using SPSS program. Covariance and Chi square test were used for the comparative analysis of different variables, considering as significant value p < 0.05. Graphics were made using HG 3.0 version. All patients gave their informed consent to participate in the study. The protocol was approved by the Ethical Committee of the Clinic Hospital. RESULTS: Systolic blood pressure (SBP) decreased from 179.00 to 156.96 mm Hg in the Lisinopril group (p < 0.001) and from 176.65 to 149.20 mm Hg in the Enalapril group (p < 0.001). Diastolic blood pressure (DBP) decreased from 104.96 to 89.94 mm Hg in the L group (p < 0.001) and from 104.16 to 86.20 mm Hg in the E group (p < 0.001). Using ABPM mean SBP daily decreased from 163.86 to 132.26 mm Hg with L (p < 0.001) and from 156.65 to 136.18 mm Hg with E (p < 0.001). Mean DBP daily decreased from 99.23 to 87.51 mm Hg with L (p < 0.001) and from 96.55 to 87.24 with E (p < 0.001). Decreases were found from total cholesterol in both groups (from 218.46 to 207.29 mg/dl with L and from 220.59 to 205.19 mg/dl with E) (p < 0.05) and Cholesterol-LDL in the E group (150.44 to 138.88) (p < 0.05). CONCLUSIONS: Both drugs produce a fall of blood pressure in mild-to-moderate hypertension, although using ABPM this fall is more significant with. According to McCall index 73.45% of patients in the Lisinopril group and 69.26% of Enalapril group controlled their hypertension, there was a difference found between BP values obtained in the clinic and those obtained using ABPM. PMID- 9221709 TI - [Predictive factors for survival and for a second coronary event in patients with a first acute myocardial infarct]. AB - Prognosis after an acute myocardial infarction is closely related to the severity of coronary obstruction, and the residual functionality of left ventricle, which may be evaluated by the ejection fraction. To evaluate the utility of the ejection fraction and some cardiovascular risk factors, as predictors of a second myocardial infarction and delayed death, in those patients with a first acute myocardial infarction, 161 hospitalized patients were included in the study. The occurrence of a second myocardial infarction or death after the first month was evaluated. All patients were followed for 1 to 51 months, and the ejection fraction through a transthoracic echocardiogram was measured. 119 men and 42 women were included in the study, with a total of 3802 person-months of follow up. The incidence rate for a second myocardial infarction was 0.01052 month,-1 and the mortality rate was 0.00342 month-1. In a Cox survival analysis model, ejection fraction was a major prognostic index and those subjects with an ejection fraction below 40% had a seven fold higher risk for a second myocardial infarction. Diabetes mellitus and hypertension were major predictors of a delayed death after a first myocardial infarction. Ejection fraction is the most related variable to the occurrence of a second myocardial infarction, and together with a history of diabetes and hypertension are good predictors of a delayed death after a first myocardial infarction. The identification of subjects with a poor prognosis may allow to establish specific preventive measures. Ejection fraction is useful to categorize patients according to their prognosis. PMID- 9221710 TI - [Cardiac rupture in acute myocardial infarct. Presentation of 20 postmortem cases]. AB - With the advancement of the Coronary Care Units in the past three decades, there had been an important reduction in mortality secondary to arrhythmias in acute myocardial infarction (AMI): been now days, cardiogenic shock and cardiac rupture the first and second causes of in-hospital death in these patients. The purpose of this report is to know the anatomoclinical characteristics in our hospital of cardiac rupture and to look for risk factors that may be considered to diagnose at the precise time this complication that might cause sudden death secondary to hemodynamic and electromechanical changes. From 300 postmortem cases with AMI proved clinical, and by anatomopathological studies, 20 cases with cardiac rupture were obtained, among which: 11 (55%) were males with an average age of 61.7 years and 9 (45%) females, with an average age of 60 years. The following coronary risk factors were detected: systemic hypertension in 15 (75%) cases; cigarette smoking in 13 (65%) cases and diabetes mellitus in 11 (55%) cases. Long lasting or recurrent history of chest pain previous to death was present in 14 (70%) cases. Conduction disturbances were detected in 13 (65%) cases; among them, 7 (35%) had third degree heart block in whom permanent pacemaker was inserted; 4 (20%) had CRBBB and 2 (10%) ASB. The average heart weight was 478 gr. in males and 434 gr. in females. Evidence of an old MI was present in 7 (35%) cases. All patients had transmural MI. Free cardiac wall rupture was seen in 14 (70%) cases and from the ventricular septum, 6 (30%) cases. Hemopericardium was present in all cases (100%) with an average amount of 425 ml of blood. Pericarditis in 3 (15%). The average time of evolution since the beginning of the AMI until death were 4 days and the main causes of death were cardiogenic shock in 17 (85%) and congestive heart failure in 3 (15%). PMID- 9221711 TI - [Is age a limitation in coronary revascularization? Comparative group analysis]. AB - It is a well known fact that in the last decade the longevity of the population has grown, which in turn has resulted in a larger group of octogenary patients being taken to coronary artery bypass procedures. Nevertheless, older age has always been considered a predisposing factor for complications. In this paper we analyze our recent experience with octogenary patients submitted to coronary artery bypass graft surgery, comparing the morbi-mortality of this group with a comparative younger age group. We found no increase in serious complications. Thus surgery should be offered irrespective of age with acceptable survival. PMID- 9221712 TI - [The origin of scientific academies]. PMID- 9221713 TI - [Stentomania]. PMID- 9221715 TI - [How I got to study legumes]. AB - In this paper the author presents a brief account of his involvement in the study of legume seeds form a nutritional and toxicological perspective. After observing that the Venezuelan peasants ate diets which often included cooked black beans and a form of corn bread called arepas, he performed nutritional trials which led him to recognize that raw beans contained thermolabile antinutritional factors and that their proteins were complementary to those of corn. Among the antinutritional factors, he isolated a hemagglutinating fraction which later was further characterized. Based on their properties he recognized the existence of four different types of Phaseolus vulgaris cultivars. Research on the nutritive value of bean diets also got him involved in the identification of a growth factor later called vitamin B12. PMID- 9221716 TI - [Genetic improvement of legumes]. AB - Genetic improvement, a process aimed to obtain populations with a high frequency of desirable phenotypes, comprises both traditional and biotechnology approaches. Through these manipulations it is possible to address any problem, provided it is associated with a gene determined character. In the case of legume crops, some features have been changed such as yield, plant morphology, susceptibility to pathogens and other stress factors which reduce grain quality and output. In Venezuela an example of genetic improvement is represented by the domestication of Canavalia ensiformis (L) DC. working out some problems which limited its use as a crop, such as the shape and growing pattern, to obtain erect plants, and pod length to avoid its contact with the humid soil which accelerates rotting. Regarding the improvement of grain quality, genotypes have been found with low canavanine content. PMID- 9221714 TI - [Guidelines and recommendations for teaching and practice of cardiac catheterization. The working group for the establishment of guidelines in cardiac catheterization, Mexican Society of Cardiology]. PMID- 9221717 TI - [Agricultural and nutritional importance of legumes]. AB - The main ecophysiologic, agronomic and economic feature of legume plants is the development of tubercles and nodules in their apical system. Nodule formation occurs in most legume species provided a compatible type of Rhizobium bacteria is present in the soil. Nitrogen fixation in nodules renders these plants independent of nitrogen fertilizers, the most expensive of all goods in modern cereal agriculture. Considering that soils may get enriched in nitrogen through fixation in nodules and the decomposition of foliage when the aerial parts of legume plants are used as green fertilizers, only through the inclusion of legume crops within planned harvest schemes, it would be possible to achieve success in large scale production strategies. Legume crops are extensively produced in temperate climates areas in which, in addition to their use in animal nutrition, yields of 18 kg per person per year are obtained. In contrast, in the Third World countries located in tropical areas, legume production is scarce, with annual yields of 9 kg per person per year. Currently, it is proposed that the energy and protein intake should match that of the developed countries 40 years ago (i.e. 3000 Kcal and 70 g protein per day); for this, it would be necessary to have an average availability of 60 g of legume seeds per person per day. Therefore, the production of legume seeds should be increased. In addition, research aimed to study and exploit the agronomic potential of this rich botanical family should be strengthened through the formation of interdisciplinary groups. PMID- 9221718 TI - [Biographical sketch of Dr. Fernando Monckebert Barros]. PMID- 9221719 TI - [Dr. Fernando Monckeberg and the creation of the Laboratory for Pediatrics Research]. PMID- 9221720 TI - [International perspective of the role of the INTA in nutrition]. PMID- 9221721 TI - [The future of the sciences-industry interaction]. PMID- 9221723 TI - [Speech pronounced by Dr. Fernando Monckeberg in appreciation of the hommage]. PMID- 9221722 TI - [Global and regional consequences of occult hunger]. PMID- 9221724 TI - A novel valveless rotary pump for cardiac assist. AB - We describe the design and preliminary testing of a novel pump designed for cardiac assist. It is a valveless rotary pump that can generate pulsatile or nonpulsatile output. The basis of the pump design is a cylindrical cavity swept by a vane sliding in a rotating disc. During rotation, rollers at the ends of the vane are in constant contact with the internal wall of the cavity so that a fluid containing space is swept around the cavity from the inflow to the outflow ports. Bench tests showed that the output was relatively independent of the outflow resistance at pressures up to 100 mm Hg. In tests in dogs, the pump performed satisfactorily as a left ventricular assist device and as a pump for total cardiopulmonary bypass. We believe that this simple pump design has potential as a left ventricular assist device or total artificial heart and merits further investigation. PMID- 9221725 TI - [New structural motifs in alpha-helical proteins]. AB - Four novel structural motifs were found and characterized in alpha-helical proteins. One of them consists of three alpha helices and can be represented as a combination of an alpha-alpha corner and an L-shaped structure. Its second alpha helix is a part of both one of the two alpha helices of the alpha-alpha corner and one of the two L-structure helices. The second structural motif, named the ABCD unit, consists of four alpha helices A, B, C, and D. These are consecutive in sequence and arranged in space in such a manner that the helices B, C, and D form a left-handed superhelix and the helix A is located between the helices B and D and is approximately antiparallel to them. The third motif, alpha helix loop-alpha helix, consists of two alpha helices and a long connection. Its alpha helices are arranged in an approximately parallel manner and together with the long connection, form a left-handed alpha-1-alpha superhelix in space. The fourth structural motif considered is formed by four consecutive alpha helices. It is named the phi motif, because its overall shape is reminiscent of the Greek letter phi. Various variants of these motifs are analyzed on numerous examples of proteins with the known spatial structures. PMID- 9221726 TI - [Denatured transitions of the molecular chaperone GroEL from Escherichia coli]. AB - Conformational changes of oligomeric particle of GroEL chaperone from E. coli in solution were studied, which proceed during its denaturation upon the action of elevated urea concentration, temperature, and extremal pH values by the methods of CD, light scattering, scanning microcalorimetry, hydrophobic probe binding, and ATPase activity measurements. The ranges of changing the external conditions; within which GroEL retains its structure and functions, were determined. Denaturation transitions were found to be cooperative, pronounced, and irreversible. In the pH range from 6.0 to 9.6, the three-step change of the ATPase activity of GroEL was shown to occur with half-transition pH1/2 of 6.3, 8.5, and 9.3. It does not result in any essential structural changes and is probably associated with a protonation/deprotonation of amino acid residues important for the GroEL ATPase activity. PMID- 9221727 TI - [Recombinant Thermus thermophilus His6-DNA polymerase with reverse transcriptase activity]. AB - The Tth DNA polymerase genes from the thermophilic bacterium Thermus thermophilus (strans HB-8 and Tt-111) were cloned and expressed in Escherichia coli cells. The target protein contained a polyhistidine tag at the N-terminus that allowed its single-stage isolation by affinity chromatography on Ni-NTA-agarose. The isolated enzyme displayed both high DNA polymerase and reverse transcriptase activities. PMID- 9221728 TI - [Photomodification of DNA with a perfluorylazide-derived oligonucleotide]. AB - The kinetics of photomodification of oligodeoxyribonucleotide pd(GTGTGA) with a derivative of the complementary oligodeoxyribonucleotide pd(CACACA) bearing a 3 (n-azidotetrafluorobenzoylamino)propylamine residue (ArN3) at the terminal phosphate group was studied at 20 degrees C. It was found that the target's G3 residue is preferentially modified. Along with the transformation of the arylazide moiety, degradation of the oligonucleotide fragment of the reagent occurred with a partial loss of affinity. With the use of the reagent labeled at the 5'-end, sites of photomodification were found. From the dependence of the modification level on the reagent concentration at the initial time of irradiation, the association constant (Kx = (1.40 +/- 0.24) x 10(5) M-1) was determined. From the dependence of the modification level on the concentration of the pre-irradiated reagent, the constant of the transformed reagent-target association in solution (Kr = (2.49 +/- 0.30) x 10(4) M-1) was determined. From the time-dependence of the modification level [PZ]/P0, the rate constant for the limiting step of photomodification (k0 = (5.31 +/- 0.28) x 10(-4) S-1) and the modification efficiency of the target in the complex with the reagent (gamma = 1) were found. PMID- 9221729 TI - [Oxidation in liposomes from egg phosphatidylcholine loaded with L-3,4 dihydroxyphenylalanine (DOPA) and dopamine: mutual effect of components]. AB - Oxidation of egg phosphatidylcholine and catechols (L-3,4-dihydroxyphenylalanine or dopamine) in liposome dispersions was studied. These catechols, encapsulated in liposomes, were shown to decrease the rate of phosphatidylcholine oxidation by factors of 4.9 and 2.6, respectively, at storage in darkness at 4 degrees C. After 40 min of Fe(II)-ascorbate-induced oxidation, the level of malonaldehyde in such liposomes remained constant, while in the empty liposomes, it increased 3,4 fold. Egg phosphatidylcholine protected catechols encapsulated into liposomes from oxidation, and the rate of this oxidation was shown to be substantially lower than that in micellar solutions under the same conditions. Evidently, this can be due to the barrier function of a lipid bilayer. PMID- 9221730 TI - [Formation of hemichrome during reaction of hemoglobin with polar phosphatidylcholine derivatives]. AB - Polar phosphatidylcholine derivatives [1-acyl-2-lyso-sn-glycero-3-phosphocholine, 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine (platelet-activating factor), 1-acyl 2-glutaryl-sn-glycero-3-phosphocholine, and 1-acyl-2-azelaoyl-sn-glycero-3 phosphocholine], which are formed in biological structures by enzymatic and free radical reactions, were studied as effectors of the conversion of methemoglobin and oxyhemoglobin into an oxidized low-spin form referred to as hemichrome. It is shown that all these phosphatidylcholine derivatives act as effectors in the course of the transition of met- and oxyhemoglobin to hemichrome. Among the compounds studied, phosphatidylcholine derivatives containing glutaric and azelaic acids residues have the greatest effect on the rate of hemichrome formation. PMID- 9221731 TI - [Affinity of 3-beta-(2-hydroxyethoxy)-5-alpha-cholest-8(14)-ene-15-one to oxysterol binding protein and its metabolism in HepG2 hepatoma cells]. AB - beta-(2-Hydroxyethoxy)-5 alpha-cholest-8(14)-en-15-one, a synthetic inhibitor of cholesterol biosynthesis, was shown to exhibit a high affinity to oxysterol binding protein. This was proved by ultracentrifugation of the protein fraction from rabbit liver in the presence of the 3H-labeled inhibitor, 3 beta-(2-hydroxy 2-[3H]ethoxy)-5 alpha-cholest-8(14)-en-15-one, or by the substitution of the [3H] 25-hydroxycholesterol in its complex with the oxysterol binding protein. In human hepatoma Hep G2 cells, the inhibitor decreased activity of 3-hydroxy-3 methylglutaryl CoA reductase [ID50 (2.7 +/- 0.7) x 10(-5) M] and was transformed into 3 beta-[2-(9-Z-octadecenoyloxy)ethoxy]-5 alpha-cholest-8(14)-en-15-one. PMID- 9221733 TI - Ticks (Acari: Ixodida) uncommonly found biting humans in North Carolina. AB - Collection records are presented that document human-biting by Otobius megnini, Amblyomma maculatum, Haemaphysalis leporispalustris, Ixodes cookei, Ixodes dentatus, and Rhipicephalus sanguineus in North Carolina. These species are either extremely rare in North Carolina or they are normally considered non-human feeders. The record of Otobius megnini represents the first collection of this species in North Carolina in over 50 years. It is proposed that immature Rhipicephalus sanguineus feed on humans much more than previously suspected and that they represent a threat for the transmission of pathogens to humans. PMID- 9221732 TI - Dispersal behavior of adult snow melt mosquitoes in the Upper Rhine Valley, Germany. AB - The dispersal behavior of female snow melt mosquitoes was studied in two forests in Rhineland-Palatinate, Germany, from April to August 1993. Both CDC-light-traps and human bait collections were used to collect mosquitoes. Sampling sites were chosen along a west-east and a north-south transect in treated and untreated parts of a forest with a village in its center. Around this settlement, breeding sites within a radius of 1.5 to 2.5 km were treated. It could be shown that this buffer zone is sufficient to prevent a nuisance caused by snow melt mosquitoes in the village. The results lead to the conclusion that snow melt mosquitoes do not regularly migrate over large distances but stay near their breeding sites. In a detailed study of the behavior of Aedes rusticus, it could be observed that these mosquitoes were resting in the interior of the forest during daytime and leaving it with increasing dusk up to 50 m from the forest edge. A comparison of landing rate counts near a row of trees and in the open field showed higher activity near the row of trees indicating visual orientation of the mosquitoes. Although the Ae. rusticus females left the forest regularly, no nuisance occurred in nearby villages. The treatment of breeding sites near settlements appeared to be sufficient to prevent a nuisance caused by the snow melt mosquitoes. PMID- 9221734 TI - The fleas (Siphonaptera) of Tennessee. AB - Thirty-three species of fleas are recorded from the state of Tennessee. New state records are reported for two species, the pulicid fleas Euhoplopsyllus glacialis affinis and Pulex simulans. Two species of fleas with catholic feeding habits appear to be especially widespread and abundant in Tennessee. These are the pulicid Ctenocephalides felis which parasitizes cats, dogs, humans, opossums, and other medium to large sized mammals, and the hystrichopsyllid Ctenophthalmus pseudagyrtes which is associated with several species of small mammals, particularly shrews, moles, voles, and native mice. For a southeastern state, Tennessee has a relatively rich flea fauna. The figure of 33 flea species recorded here for Tennessee is higher than documented figures for other southeastern states (17 species for Alabama, 19 for Florida, 20 for Georgia, 12 for Mississippi, 18 for North Carolina, 19 for South Carolina). This is largely because several species with boreal origins inhabit the higher elevations characteristic of the Appalachian Mountains in the eastern part of the state. Although plague is not enzootic as far east as Tennessee, and murine typhus is rare of absent, suitable flea vectors inhabit the state and one abundant flea species, C. felis, is a pest because it feeds on companion animals and humans. PMID- 9221735 TI - Culicidae (Diptera) in the diet of predatory stages of anurans (Amphibia) in humid biotopes of the Rhine Valley in Germany. AB - A three-year field study was conducted during 1993 to 1995 to determine the importance of mosquitoes in the diet of anurans. The study was aimed to assess the impact of biological mosquito control on the populations of amphibia in the Rhine Valley, Germany. Sampling took place in two areas with stands of different deciduous trees at the western bank of the Rhine (north of Karlsruhe, Germany) from early May to late October. The frequency and species composition of the terrestrial stages of Amphibia was monitored by hand catches and by live pitfall trapping. A total of 2,419 Amphibia were caught in the three years. Of these, 95.8% were anurans, consisting of 77% Ranidae, with 25.5% Rana arvalis, and 4.2% were Urodela, Salamandridae. All anura caught were subjected to "stomach flushing" to yield their stomach contents before they were released again. The stomachs of 2,163 anuran specimens contained an average of 7.7 prey items, of which only 0.16% were Culicidae. In R. arvalis the total diet consisted of 33% Collembola, spiders and beetles; 0.1% of the specimens in the diet were Culicidae. The most common culicid species in the study area, Aedes vexans, was also most often found in the anuran stomachs. However, no correlation existed between the number of mosquitoes and their number as prey of Anura. It is concluded that anurans will not be negatively affected by biological mosquito control in the Rhine Valley. Furthermore, the impact of anurans on Culicidae seems to be negligible. PMID- 9221736 TI - Organic enrichment of breeding sources for sustained productivity of mosquitoes (Diptera: Culicidae). AB - Alfalfa pellets and chicken lay mash were used to obtain polluted water for continuous and sustained oviposition by Culex mosquitoes. Water in fiberglass tubs enriched with 0.1 and 0.25 percent (w/v) of either material received heavy oviposition by gravid mosquitoes. The higher level of enrichment resulted in greater oviposition than lower concentrations which in turn received significantly more oviposition than the controls with no enrichment. Oviposition continued in the enriched tubs for more than 43 days where Culex quinquefasciatus and Culex stigmatosoma constituted a major proportion of the eggs laid while Culex tarsalis, as expected, oviposited at much lower rates than the other two species. Supplemental enrichment of mosquito breeding sources with organic material can thus provide uniform and sustained availability of some mosquito larvae for the evaluation of persistent control agents and other related studies. PMID- 9221737 TI - Infection and transmission of Plasmodium gallinaceum (Eucoccida: Plasmodiidae) in Aedes aegypti (Diptera: Culicidae): effect of preinfection sugar meals and postinfection blood meals. AB - Frequency of sugar feeding and blood feeding can have an impact on the infection by and transmission of malaria parasites. Data presented here indicate that frequent blood feeding has a deleterious effect on infection by malaria parasites in Aedes aegypti. In addition, mosquitoes that do not blood feed, but instead feed on sugar alone after an infected blood meal, have a higher rate of parasite transmission than mosquitoes fed additional blood meals. PMID- 9221738 TI - Efficacy of a granular formulation of Bacillus sphaericus against Culex quinquefasciatus and Anopheles gambiae in West African countries. AB - The efficacy of a sustained released granular formulation of Bacillus sphaericus strain 2362 was compared to a flowable concentrate in containers, cesspools, and water ponds. Duration of control was dependent on formulation, dosage, exposure to sun, site, recycling, and target mosquito larvae. In a series of container tests with repeated additions of fourth-instar Culex quinquefasciatus larvae exposed to 0.3 or 3.0 g/m2 when cadavers were not removed, more than 95 percent control was obtained for two and four days in containers that were exposed to the sun with sewage water treated with the flowable concentrate compared to four and seven days for those treated with the granule. In sun-exposed containers with sewage water, control persisted for two days for the flowable concentrate at both dosages and one and six days for the granule at 0.3 g/m2 and 3.0 g/m2, respectively. Compared to the above tests, more than seven weeks control was obtained with 0.3 g/m2 of the flowable concentrate in closed containers where larvae were added weekly. In closed containers without weekly addition of larvae, the control was 15 percent when larvae were added five weeks after the treatment. Spore counts at the surface and bottom of the containers with lids showed an increase in number of spores at the surface where larvae were added weekly and a rapid decline where they were not. Spore counts at the bottom showed settling in both cases, but to a much higher level where larvae were added weekly. Nearly 100 percent control of Cx, quinquefasciatus larvae was obtained for at least 16 days in cesspools in Yaounde, Cameroon, treated with the granule at 3.0 g/m2 compared to approximately nine days for the flowable concentrate. At 0.3 g/m2, the duration of this reduction was five days for both products. Nearly 100 percent control of Anopheles gambiae was obtained in sun-exposed water ponds near the village Kotiokh, Senegal, for at least 15 days with the granule at 3.0 g/m2 compared to just five days for the flowable concentrate at the same dosage. PMID- 9221740 TI - Diel oviposition patterns of Aedes albopictus (Skuse) and Aedes triseriatus (Say) in the laboratory and the field. AB - The oviposition patterns of Aedes albopictus and Aedes triseriatus were observed in preliminary field experiments during the summer of 1995 and in the laboratory the following winter. Aedes albopictus exhibited a diel periodicity of oviposition in the field, ovipositing a significantly greater number of eggs during the day than during the night (P = 0.0001). Laboratory observations for 40 consecutive hours indicated that Ae. albopictus oviposited only during the hours of light, with a broad peak of oviposition activity occurring in mid-afternoon. Aedes triseriatus, however, oviposited during all periods of the day and night in the field. A significantly greater number of eggs were oviposited in traps open 24 hours than in traps open only during the day (P = 0.01), whereas there was no significant difference in the number of eggs deposited in traps open 24 hours and those open only during the night (P = 0.14). In the laboratory, Ae. triseriatus oviposited during all periods of light and dark, with a distinct peak of oviposition activity occurring during the evening crepuscular period. PMID- 9221739 TI - Community structure and prevalence of hantavirus infection in rodents: a geographic division of the enzootic area in far eastern Russia. AB - The results of an extensive rodent trapping effort throughout the southern part of far eastern Russia and hantavirus antigen screening of tissues were used to develop a multifaceted approach for the geographic division of the enzootic territory of hantavirus. Four species of rodents (Apodemus agrarius, Apodemus peninsulae, Microtus fortis, and Clethrionomys rufocanus) comprised 88.5 percent of 10,595 captured rodents and 94.1 percent of 996 antigen-positive animals. Rodent fauna and the prevalence and distribution of hantavirus antigen-positive animals were compared among major biotic communities in the region. The species composition of the rodent communities and the predominant hantavirus reservoir species were used as criteria to define zones with similar enzootic characteristics. PMID- 9221741 TI - Sucking lice (Anoplura) of mammals of Tennessee. AB - Twenty-five species of sucking lice are recorded from wild and domestic mammals, including humans, from Tennessee. Collections of 10 of these species (Haematopinus eurysternus, Hoplopleura captiosa, Hoplopleura hirsuta, Hoplopleura oryzomydis, Hoplopleura trispinosa, Linognathus africanus, Linognathus setosus, Linognathus vituli, Neohaematopinus sciuropteri and Polyplax auricularis) represent newly documented state records. Host specificity was exhibited by 22 species of lice with each of these species being recovered from just one mammal species. Louse infestation prevalences are included for large samples of hosts. A host-parasite list for Tennessean sucking lice is included. PMID- 9221742 TI - An overview of plague in the United States and a report of investigations of two human cases in Kern county, California, 1995. AB - Plague was confirmed in the United States from nine western states during 1995. Evidence of Yersinia pestis infection was identified in 28 species of wild or domestic mammals. Thirteen of the plague positive species were wild rodents; 15 were predators/carnivores. Yersinia pestis was isolated from eight species of fleas. Seven confirmed cases of human plague were reported in 1995 (New Mexico 3; California 2; Arizona and Oregon 1 each). Five of the seven cases were bubonic; one was septicemic and one a fatal pneumonic case. Months of onset ranged from March through August. In California, during 1995, plague was recorded from 15 of the 58 counties. Over 1,500 animals were tested, of which 208 were plague positive. These included 144 rodents and 64 predators/carnivores. Two confirmed human cases (one bubonic and one fatal pneumonic) occurred, both in Kern County. Case No. 1 was reported from the town of Tehachapi. The patient, a 23 year-old male resident, died following a diagnosis of plague pneumonia. The patient's source of plague infection could not be determined precisely. Field investigations revealed an extensive plague epizootic surrounding Tehachapi, an area of approximately 500-600 square miles (800-970 square kilometers). Case No. 2 was a 57 year-old female diagnosed with bubonic plague; she was placed on an antibiotic regimen and subsequently recovered. The patient lives approximately 20 miles (32 km) north of Tehachapi. Field investigations revealed evidence of a plague epizootic in the vicinity of the victim's residence and adjacent areas. Overall results of the joint field investigations throughout the entire Kern county area revealed a high rate of plague positive animals. Of the numerous samples submitted, 48 non-human samples were plague positive. PMID- 9221743 TI - Energetics and sugar-feeding of field-collected anopheline females. AB - We studied the relationship between nutritional reserves and blood-feeding and sugar-feeding of Anopheles freeborni (Diptera: Culicidae) females in the field. In particular we determined whether (1) females feed on nectar before maturing eggs and initiating host-seeking and (2) the energy reserves of host-seeking females differ from those of non-fed resting females. Twenty-three percent of host-seeking females and 94 percent of gravid females were positive for nectar sugars (containing > 20 micrograms of fructose) versus 55 percent of empty (no blood or eggs) females collected in the morning and 36 percent of empty females collected in the evening. In addition, gravid females contained significantly more calories of nectar than empty, blood-fed, or partially blood-fed females collected in the morning. When the energy reserves of host-seeking and resting females were compared, no differences were found in lipid, trehalose, or glycogen. However, empty females collected in the evening contained more glycogen than empty females collected in the morning. We conclude that gravid females frequently feed on nectar and that fructose is metabolized into glycogen during the day. PMID- 9221744 TI - Larval competition in Aedes triseriatus (Diptera: Culicidae): effects of density on size, growth, sex ratio, and survival. AB - We assessed the effect of increased larval density on selected life table attributes of larval Aedes triseriatus. Larvae were reared at densities of 0.125, 0.250, and 0.375 larvae/cm3 of water. The parameters we evaluated included duration of each developmental instar, percent survival to adult emergence, sex ratio, and size of early and late emerging adults. The amount of time spent in the first and second instar was not affected by density, but crowding lengthened duration of later instars significantly. Males emerged earlier than females at all densities, and there was no difference in the ratio of males to females due to crowding. Crowding did increase the overall developmental time of both sexes and produced significantly smaller adults of both sexes. Densities of 0.375 larvae/cm3 resulted in a significant increase in mortality. Results suggest that the effects of crowding in container habitats should be allowed for in the construction of time-specific life tables where duration of larval instars is used in the calculations. The importance of smaller body size as a result of crowding in container habitats is discussed in terms of vector competence of Ae. triseriatus in the transmission of LaCrosse encephalitis virus. PMID- 9221745 TI - Reduced transfer of male accessory gland proteins and monandry in female Aedes aegypti mosquitoes. AB - We examined whether female Aedes aegypti (L.) mosquitoes would remate after they first mated with a male that was reared on a suboptimal larval diet and that as a result, transferred reduced amounts of male accessory gland proteins. Accessory gland proteins from males labeled with 3H leucine were not detected in females allowed to male mate with the labeled males after they first mated with either low diet or high diet males. The amount of the male accessory gland protein transferred by smaller, low diet males was adequate to terminate female receptivity, even after one gonotrophic cycle, and females of this species appear to be monogamous. PMID- 9221746 TI - Identification of a novel developmental stage marking lineage commitment of progenitor thymocytes. AB - Bipotent progenitors for T and natural killer (NK) lymphocytes are thought to exist among early precursor thymocytes. The identification and functional properties of such a progenitor population remain undefined. We report the identification of a novel developmental stage during fetal thymic ontogeny that delineates a population of T/NK-committed progenitors (NK1. 1(+)/CD117(+)/CD44(+)/CD25(-)). Thymocytes at this stage in development are phenotypically and functionally distinguishable from the pool of multipotent lymphoid-restricted (B, T, and NK) precursor thymocytes. Exposure of multipotent precursor thymocytes or fetal liver- derived hematopoietic progenitors to thymic stroma induces differentiation to the bipotent developmental stage. Continued exposure to a thymic microenvironment results in predominant commitment to the T cell lineage, whereas coculture with a bone marrow-derived stromal cell line results in the generation of mature NK cells. Thus, the restriction point to T and NK lymphocyte destinies from a multipotent progenitor stage is marked by a thymus-induced differentiation step. PMID- 9221747 TI - Protective immunity to nematode infection is induced by CTLA-4 blockade. AB - The recent observation that neutralization or genetic deletion of the T lymphocyte receptor CTLA-4 allows enhanced T cell reactivity offers new opportunities for immunotherapy against infectious agents. We used a neutralizing antibody to block CTLA-4 interaction with its ligands CD80 and CD86 during infection of mice with the nematode, Nippostrongylus brasiliensis. CTLA-4 blockade greatly enhanced and accelerated the T cell immune response to N. brasiliensis, resulting in a profound reduction in adult worm numbers and early termination of parasite egg production. The ability of CTLA-4 blockade to accelerate primary immune responses to a protective level during an acute infection indicates its potential as an immunotherapeutic tool for dealing with infectious agents. PMID- 9221748 TI - Tumor necrosis factor alpha and interleukin 1beta enhance the cortisone/cortisol shuttle. AB - Endogenously released or exogenously administered glucocorticosteroids are relevant hormones for controlling inflammation. Only 11beta-hydroxy glucocorticosteroids, but not 11-keto glucocorticosteroids, activate glucocorticoid receptors. Since we found that glomerular mesangial cells (GMC) express 11beta-hydroxysteroid dehydrogenase 1 (11beta-OHSD1), which interconverts 11-keto glucocorticosteroids into 11beta-hydroxy glucocorticosteroids (cortisone/cortisol shuttle), we explored whether 11beta-OHSD1 determines the antiinflammatory effect of glucocorticosteroids. GMC exposed to interleukin (IL) 1beta or tumor necrosis factor alpha (TNF-alpha) release group II phospholipase A2 (PLA2), a key enzyme producing inflammatory mediators. 11beta-hydroxy glucocorticosteroids inhibited cytokine-induced transcription and release of PLA2 through a glucocorticoid receptor-dependent mechanism. This inhibition was enhanced by inhibiting 11beta-OHSD1. Interestingly, 11-keto glucocorticosteroids decreased cytokine-induced PLA2 release as well, a finding abrogated by inhibiting 11beta-OHSD1. Stimulating GMC with IL-1beta or TNF-alpha increased expression and reductase activity of 11beta-OHSD1. Similarly, this IL-1beta- and TNF-alpha-induced formation of active 11beta-hydroxy glucocorticosteroids from inert 11-keto glucocorticosteroids by the 11beta-OHSD1 was shown in the Kiki cell line that expresses the stably transfected bacterial beta-galactosidase gene under the control of a glucocorticosteroids response element. Thus, we conclude that 11beta-OHSD1 controls access of 11beta-hydroxy glucocorticosteroids and 11 keto glucocorticosteroids to glucocorticoid receptors and thus determines the anti-inflammatory effect of glucocorticosteroids. IL-1beta and TNF-alpha upregulate specifically the reductase activity of 11beta-OHSD1 and counterbalance by that mechanism their own proinflammatory effect. PMID- 9221749 TI - The human C3a receptor is expressed on neutrophils and monocytes, but not on B or T lymphocytes. AB - The pathophysiological relevance of the complement split product C3a as a proinflammatory mediator is still ill defined. The expression pattern of the human C3a receptor (C3aR) can provide important clues for the role of this anaphylatoxin in inflammation. There is strong evidence for C3aR expression on basophils, and eosinophils, but additionally, only on tumor cell lines of leukemic or hepatic origin. It is unclear whether neutrophils also express the C3aR, but need a costimulus provided by eosinophils for certain biological responses, or whether neutrophils lack the C3aR and respond to C3a via a secondary stimulus generated by eosinophils, i.e., by an indirect mode. In the present study, polyclonal antiserum raised against the second extracellular loop of the C3aR was used to characterize C3aR expression on peripheral blood leukocytes. For high degree purification of neutrophils, a negative selection method was established that decreased the contamination with CD9(bright+) eosinophils down to <0.2%. Flow cytometric analyses, functional assays, and binding assays on highly purified neutrophils confirmed C3aR expression and coupling. Monocytes were identified as an additional C3aR-positive cell population of the peripheral blood. The expression of the C3aR on eosinophils could be confirmed. In contrast, the receptor could not be detected on unchallenged B or T lymphocytes (or lymphocyte-derived Raji cells). PMID- 9221751 TI - Itk negatively regulates induction of T cell proliferation by CD28 costimulation. AB - CD28 is a cell surface molecule that mediates a costimulatory signal crucial for T cell proliferation and lymphokine production. The signal transduction mechanisms of CD28 are not well understood. Itk, a nonreceptor protein tyrosine kinase specifically expressed in T cells and mast cells, has been implicated in the CD28 signaling pathway because of reports that it becomes phosphorylated on tyrosines and associates with CD28 upon cross-linking of the cell surface molecule. To determine whether Itk plays a functional role in CD28 signaling, we compared T cells from Itk-deficient mice and control mice for their responses to CD28 costimulation. T cells defective in Itk were found to be fully competent to respond to costimulation. Whereas the CD3-mediated proliferative response was severely compromised in the absence of Itk, the calcineurin-independent CD28 mediated response was significantly elevated when compared with cells from control animals. The augmented proliferation was not due to increased production of interleukin-2. The results suggest that Itk has distinct roles in the CD3 versus the CD28 signaling pathways. By negatively regulating the amplitude of signaling upon CD28 costimulation, Itk may provide a means for modulating the outcome of T cell activation during development and during antigen-driven immune responses. PMID- 9221750 TI - Potential immunocompetence of proteolytic fragments produced by proteasomes before evolution of the vertebrate immune system. AB - To generate peptides for presentation by major histocompatibility complex (MHC) class I molecules to T lymphocytes, the immune system of vertebrates has recruited the proteasomes, phylogenetically ancient multicatalytic high molecular weight endoproteases. We have previously shown that many of the proteolytic fragments generated by vertebrate proteasomes have structural features in common with peptides eluted from MHC class I molecules, suggesting that many MHC class I ligands are direct products of proteasomal proteolysis. Here, we report that the processing of polypeptides by proteasomes is conserved in evolution, not only among vertebrate species, but including invertebrate eukaryotes such as insects and yeast. Unexpectedly, we found that several high copy ligands of MHC class I molecules, in particular, self-ligands, are major products in digests of source polypeptides by invertebrate proteasomes. Moreover, many major dual cleavage peptides produced by invertebrate proteasomes have the length and the NH2 and COOH termini preferred by MHC class I. Thus, the ability of proteasomes to generate potentially immunocompetent peptides evolved well before the vertebrate immune system. We demonstrate with polypeptide substrates that interferon gamma induction in vivo or addition of recombinant proteasome activator 28alpha in vitro alters proteasomal proteolysis in such a way that the generation of peptides with the structural features of MHC class I ligands is optimized. However, these changes are quantitative and do not confer qualitatively novel characteristics to proteasomal proteolysis. The data suggest that proteasomes may have influenced the evolution of MHC class I molecules. PMID- 9221752 TI - Antigenic cancer cells grow progressively in immune hosts without evidence for T cell exhaustion or systemic anergy. AB - One enigma in tumor immunology is why animals bearing malignant grafts can reject normal grafts that express the same nonself-antigen. An explanation for this phenomenon could be that different T cell clones react to the normal graft and the malignant cells, respectively, and only the tumor-reactive clonotypes may be affected by the growing tumor. To test this hypothesis, we used a T cell receptor transgenic mouse in which essentially all CD8(+) T cells are specific for a closely related set of self-peptides presented on the MHC class I molecule Ld. We find that the tumor expressed Ld in the T cell receptor transgenic mice but grew, while the Ld-positive skin was rejected. Thus, despite an abundance of antigen specific T cells, the malignant tissue grew while normal tissue expressing the same epitopes was rejected. Therefore, systemic T cell exhaustion or anergy was not responsible for the growth of the antigenic cancer cells. Expression of costimulatory molecules on the tumor cells after transfection and preimmunization by full-thickness skin grafts was required for rejection of a subsequent tumor challenge, but there was no detectable effect of active immunization once the tumor was established. Thus, the failure of established tumors to attract and activate tumor-specific T cells at the tumor site may be a major obstacle for preventive or therapeutic vaccination against antigenic cancer. PMID- 9221753 TI - Class I-restricted cross-presentation of exogenous self-antigens leads to deletion of autoreactive CD8(+) T cells. AB - In this report, we show that cross-presentation of self-antigens can lead to the peripheral deletion of autoreactive CD8(+) T cells. We had previously shown that transfer of ovalbumin (OVA)-specific CD8(+) T cells (OT-I cells) into rat insulin promoter-membrane-bound form of OVA transgenic mice, which express the model autoantigen OVA in the proximal tubular cells of the kidneys, the beta cells of the pancreas, the thymus, and the testis of male mice, led to the activation of OT-I cells in the draining lymph nodes. This was due to class I-restricted cross presentation of exogenous OVA on a bone marrow-derived antigen presenting cell (APC) population. Here, we show that adoptively transferred or thymically derived OT-I cells activated by cross-presentation are deleted from the peripheral pool of recirculating lymphocytes. Such deletion only required antigen recognition on a bone marrow-derived population, suggesting that cells of the professional APC class may be tolerogenic under these circumstances. Our results provide a mechanism by which the immune system can induce CD8(+) T cell tolerance to autoantigens that are expressed outside the recirculation pathway of naive T cells. PMID- 9221754 TI - Neisseria gonorrhoeae epithelial cell interaction leads to the activation of the transcription factors nuclear factor kappaB and activator protein 1 and the induction of inflammatory cytokines. AB - We have studied the effect of human bacterial pathogen Neisseria gonorrhoeae (Ngo) on the activation of nuclear factor (NF)-kappaB and the transcriptional activation of inflammatory cytokine genes upon infection of epithelial cells. During the course of infection, Ngo, the etiologic agent of gonorrhea, adheres to and penetrates mucosal epithelial cells. In vivo, localized gonococcal infections are often associated with a massive inflammatory response. We observed upregulation of several inflammatory cytokine messenger RNAs (mRNAs) and the release of the proteins in Ngo-infected epithelial cells. Moreover, infection with Ngo induced the formation of a NF-kappaB DNA-protein complex and, with a delay in time, the activation of activator protein 1, whereas basic leucine zipper transcription factors binding to the cAMP-responsive element or CAAT/enhancer-binding protein DNA-binding sites were not activated. In supershift assays using NF-kappaB-specific antibodies, we identified a NF-kappaB p50/p65 heterodimer. The NF-kappaB complex was formed within 10 min after infection and decreased 90 min after infection. Synthesis of tumor necrosis factor alpha and interluekin (IL)-1beta occurred at later times and therefore did not account for NF-kappaB activation. An analysis of transiently transfected IL-6 promoter deletion constructs suggests that NF-kappaB plays a crucial role for the transcriptional activation of the IL-6 promoter upon Ngo infection. Inactivation of NF-kappaB conferred by the protease inhibitor N-tosyl--phenylalanine chloromethyl ketone inhibited mRNA upregulation of most, but not all, studied cyctokine genes. Activation of NF-kappaB and cytokine mRNA upregulation also occur in Ngo-infected epithelial cells that were treated with cytochalasin D, indicating an extracellular signaling induced before invasion. PMID- 9221755 TI - Differential effects of B cell receptor and B cell receptor-FcgammaRIIB1 engagement on docking of Csk to GTPase-activating protein (GAP)-associated p62. AB - The stimulatory and inhibitory pathways initiated by engagement of stimulatory receptors such as the B cell receptor for antigen (BCR) and inhibitory receptors such as Fcgamma receptors of the IIB1 type (FcgammaRIIB1) intersect in ways that are poorly understood at the molecular level. Because the tyrosine kinase Csk is a potential negative regulator of lymphocyte activation, we examined the effects of BCR and FcgammaRIIB1 engagement on the binding of Csk to phosphotyrosine containing proteins. Stimulation of a B lymphoma cell line, A20, with intact anti IgG antibody induced a direct, SH2-mediated association between Csk and a 62-kD phosphotyrosine-containing protein that was identified as RasGTPase-activating protein-associated p62 (GAP-A.p62). In contrast, stimulation of A20 cells with anti-IgG F(ab')2 resulted in little increase in the association of Csk with GAP A.p62. The effect of FcgammaRIIB1 engagement on this association was abolished by blockade of FcgammaRIIB1 with the monoclonal antibody 2.4G2. Furthermore, the increased association between Csk and GAP-A.p62 seen upon stimulation with intact anti-Ig was abrogated in the FcgammaRIIB1-deficient cell line IIA1.6 and recovered when FcgammaRIIB1 expression was restored by transfection. The differential effects of BCR and BCR-FcgammaRIIB1-mediated signaling on the phosphorylation of GAP-A.p62 and its association with Csk suggest that docking of Csk to GAP-A.p62 may function in the negative regulation of antigen receptor mediated signals in B cells. PMID- 9221756 TI - Measles virus nucleocapsid protein binds to FcgammaRII and inhibits human B cell antibody production. AB - Despite the development of an efficient specific immune response during measles virus (MV) infection, an immunosuppression occurs contributing to secondary infections. To study the role of nucleocapsid protein (NP) in MV-induced immunosuppression, we produced recombinant MV NP. Purified recombinant NP exhibited biochemical, antigenic, and tridimensional structure similar to viral NP. By flow cytometry, we showed that viral or recombinant NP bound to human and murine B lymphocytes, but not to T lymphocytes. This binding was specific, independent of MHC class II expression, and dependent of the B lymphocyte activation state. The murine IIA1. 6 B cell line, deficient in the Fc receptor for IgG (FcgammaRII) expression, did not bind NP efficiently. Transfected IIA1.6 cells expressing either murine FcgammaRIIb1 or b2, or human FcgammaRIIa, b1*, or b2 isoforms efficiently bound NP. Furthermore, this binding was inhibited up to 90% by monoclonal antibodies 2.4G2 or KB61 specific for murine and human FcgammaRII, respectively. Finally, the in vitro Ig synthesis of CD40- or Ig activated human B lymphocytes in the presence of interleukin (IL)-2 and IL-10 was reduced by 50% in the presence of recombinant NP. These data demonstrate that MV NP binds to human and murine FcgammaRII and inhibits in vitro antibody production, and therefore suggests a role for NP in MV-induced immunosuppression. PMID- 9221757 TI - IkappaBalpha overexpression delays tumor formation in v-rel transgenic mice. AB - We have previously shown that transgenic mice expressing the oncoprotein v-Rel under the control of a T cell-specific promoter develop T cell lymphomas. Tumor formation was correlated with the presence of p50/v-Rel and v-Rel/v-Rel nuclear kappaB-binding activity. Since experimental evidence has led to the suggestion of a potential tumor suppressor activity for IkappaBalpha, we have studied the role of IkappaBalpha in the transforming activity of v-Rel by overexpressing IkappaBalpha in v-rel transgenic mice. Overexpression of IkappaBalpha in v-rel transgenic mice resulted in an extended survival, and the development of cutaneous T cell lymphomas of CD8(+)CD4(-) phenotype. These phenotypic alterations were associated with a dramatic reduction of p50/v-Rel, but not v Rel/v-Rel nuclear DNA binding activity and an increased expression of the intercellular adhesion molecule 1. Our results indicate that v-Rel homodimers are active in transformation and that the capacity of v-Rel-containing complexes to escape the inhibitory effect of IkappaBalpha may be a key element in its transforming capability. PMID- 9221758 TI - Identification of a thymic epithelial cell subset sharing expression of the class Ib HLA-G molecule with fetal trophoblasts. AB - HLA-G is the only class I determinant of the major histocompatibility complex (MHC) expressed by the trophoblasts, the fetal cells invading the maternal decidua during pregnancy. A unique feature of this nonclassical HLA molecule is its low polymorphism, a property that has been postulated to play an important role in preventing local activation of maternal alloreactive T and natural killer cells against the fetus. Yet, the mechanisms by which fetal HLA-G can be recognized as a self-MHC molecule by the maternal immune system remain unclear. Here we report the novel observation that HLA-G is expressed in the human thymus. Expression is targeted to the cell surface of thymic medullary and subcapsular epithelium. Thymic epithelial cell lines were generated and shown to express three alternatively spliced HLA-G transcripts, previously identified in human trophoblasts. Sequencing of HLA-G1 transcripts revealed a few nucleotide changes resulting in amino acid substitutions, all clustered within exon 3 of HLA-G, encoding for the alpha2 domain of the molecule. Our findings raise the possibility that maternal unresponsiveness to HLA-G-expressing fetal tissues may be shaped in the thymus by a previously unrecognized central presentation of this MHC molecule on the medullary epithelium. PMID- 9221759 TI - T helper 2 (Th2) T cells induce acute pancreatitis and diabetes in immune compromised nonobese diabetic (NOD) mice. AB - Autoimmune diabetes is caused by the CD4(+), T helper 1 (Th1) cell-mediated apoptosis of insulin-producing beta cells. We have previously shown that Th2 T cells bearing the same T cell receptor (TCR) as the diabetogenic Th1 T cells invade islets in neonatal nonobese diabetic (NOD) mice but fail to cause disease. Moreover, when mixed in excess and cotransferred with Th1 T cells, Th2 T cells could not protect NOD neonates from Th1-mediated diabetes. We have now found, to our great surprise, the same Th2 T cells that produced a harmless insulitis in neonatal NOD mice produced intense and generalized pancreatitis and insulitis associated with islet cell necrosis, abscess formation, and subsequent diabetes when transferred into immunocompromised NOD.scid mice. These lesions resembled allergic inflamation and contained a large eosinophilic infiltrate. Moreover, the Th2-mediated destruction of islet cells was mediated by local interleukin-10 (IL 10) production but not by IL-4. These findings indicate that under certain conditions Th2 T cells may not produce a benign or protective insulitis but rather acute pathology and disease. Additionally, these results lead us to question the feasibility of Th2-based therapy in type I diabetes, especially in immunosuppressed recipients of islet cell transplants. PMID- 9221760 TI - Myelin basic protein-specific T helper 2 (Th2) cells cause experimental autoimmune encephalomyelitis in immunodeficient hosts rather than protect them from the disease. AB - Chronic inflammatory autoimmune diseases such as multiple sclerosis, diabetes, and rheumatoid arthritis are caused by CD4(+) Th1 cells. Because Th2 cells antagonize Th1 cell functions in several ways, it is believed that immune deviation towards Th2 can prevent or cure autoimmune diseases. Experimental autoimmune encephalomyelitis (EAE) is a demyelinating disease used as a model for multiple sclerosis. Using an adoptive transfer system we assessed the role of Th1 and Th2 cells in EAE. In vitro generated Th1 and Th2 cells from myelin basic protein (MBP)-specific TCR transgenic mice were transferred into normal and immunodeficient mice. Th1 cells caused EAE in all recipients after a brief preclinical phase. Surprisingly, Th2 cells also caused EAE in RAG-1 KO mice and in alphabeta T cell-deficient mice, albeit after a longer preclinical phase. Normal or gammadelta T cell-deficient mice were resistant to EAE induced by Th2 cells. The histopathological features of this disease resembled those of an allergic process. In addition, disease induction by Th1 cells was not altered by coadmininstration of Th2 cells in any of the recipients. These findings indicate that MBP-specific Th2 cells have the potential to induce EAE and that the disease induced by previously activated Th1 cells cannot be prevented by normal lymphocytes nor by previously activated Th2 cells. PMID- 9221761 TI - Human peripheral blood eosinophils express a functional c-kit receptor for stem cell factor that stimulates very late antigen 4 (VLA-4)-mediated cell adhesion to fibronectin and vascular cell adhesion molecule 1 (VCAM-1). AB - We evaluated mature peripheral blood eosinophils for their expression of the surface tyrosine kinase, c-kit, the receptor for the stromal cell-derived cytokine, stem cell factor (SCF). Cytofluorographic analysis revealed that c-kit was expressed on the purified peripheral blood eosinophils from 8 of 8 donors (4 nonatopic and 4 atopic) (mean channel fluorescence intensity 2.0- 3. 6-fold, average 2.8 +/- 0.6-fold, greater than the negative control). The uniform and selective expression of c-kit by eosinophils was confirmed by immunohistochemical analysis of peripheral blood buffy coats. The functional integrity of c-kit was demonstrated by the capacity of 100 ng/ml (5 nM) of recombinant human (rh) SCF to increase eosinophil adhesion to 3, 10, and 30 microg/ml of immobilized FN40, a 40 kD chymotryptic fragment of plasma fibronectin, in 15 min by 7.7 +/- 1.4-, 5.3 +/ 3.3-, and 5.4 +/- 0. 2-fold, respectively, and their adhesion to 0.1, 0.5, and 1.0 microg/ml vascular cell adhesion molecule-1 (VCAM-1), by 12.7 +/- 9. 2-, 3.8 +/- 2.5-, and 1.7 +/- 0.6-fold, respectively. The SCF-stimulated adhesion occurred without concomitant changes in surface integrin expression, thereby indicating an avidity-based mechanism. rhSCF (100 ng/ml, 5 nM) was comparable to rh eotaxin (200 ng/ml, 24 nM) in stimulating adhesion. Cell adhesion to FN40 was completely inhibited with antibodies against the alpha4 and beta1 integrin subunits, revealing that the SCF/c-kit adhesion effect was mediated by a single integrin heterodimer, very late antigen 4 (VLA-4). Thus, SCF represents a newly recognized stromal ligand for the activation of eosinophils for VLA-4-mediated adhesion, which could contribute to the exit of these cells from the blood, their tissue localization, and their prominence in inflammatory lesions. PMID- 9221762 TI - Interleukin 4 (IL-4) or IL-7 prevents the death of resting T cells: stat6 is probably not required for the effect of IL-4. AB - Although much is known about the activation, proliferation, and function of CD4(+) T cells, little is known about how they survive as resting T cells in animals. Resting T cells have a half-life in animals of more than a week; however, when they are removed from animals and placed in tissue culture their half-life falls to approximately 24 h. In this paper, we show that the survival of resting T cells in vitro is promoted by two cytokines, interleukins 4 and 7 (IL-4, IL-7). They may do this in part by maintaining levels of survival promoting proteins such as Bcl-2 in the cells, because the levels of Bcl-2 and Bcl-Xl in resting T cells fall rapidly after the cells are isolated from animals, and are maintained by culture in IL-4. Because the IL-4 receptor is known to signal through the JAK1 and JAK3/Stat6 pathway, we tested whether Stat6 was required for IL-4- dependent T cell survival. Surprisingly, we found that IL-4 rescued T cells from apoptosis in what appeared to be a Stat6-independent manner. These results demonstrate that the survival of resting T cells is an active process that can be affected by signals delivered by cytokines and also suggest that the IL-4 receptor on resting T cells may use a novel signaling pathway to facilitate T cell viability. PMID- 9221763 TI - Distinct roles for signals relayed through the common cytokine receptor gamma chain and interleukin 7 receptor alpha chain in natural T cell development. AB - The commitment, differentiation, and expansion of mainstream alpha/beta T cells during ontogeny depend on the highly controlled interplay of signals relayed by cytokines through their receptors on progenitor cells. The role of cytokines in the development of natural killer (NK)1(+) natural T cells is less clearly understood. In an approach to define the role of cytokines in the commitment, differentiation, and expansion of NK1(+) T cells, their development was studied in common cytokine receptor gamma chain (gammac) and interleukin (IL)-7 receptor alpha (IL-7Ralpha)-deficient mice. These mutations block mainstream alpha/beta T cell ontogeny at an early prethymocyte stage. Natural T cells do not develop in gammac-deficient mice; they are absent in the thymus and peripheral lymphoid organs such as the liver and the spleen. In contrast, NK1(+) T cells develop in IL-7Ralpha-deficient mice in the thymus, and they are present in the liver and in the spleen. However, the absolute number of NK1(+) T cells in the thymus of IL 7Ralpha-deficient mice is reduced to approximately 10%, compared to natural T cell number in the wild-type thymus. Additional data revealed that NK1(+) T cell ontogeny is not impaired in IL-2- or IL-4-deficient mice, suggesting that neither IL-2, IL-4, nor IL-7 are required for their development. From these data, we conclude that commitment and/or differentiation to the NK1(+) natural T cell lineage requires signal transduction through the gammac, and once committed, their expansion requires signals relayed through the IL-7Ralpha. PMID- 9221765 TI - Slow-channel myasthenic syndrome caused by enhanced activation, desensitization, and agonist binding affinity attributable to mutation in the M2 domain of the acetylcholine receptor alpha subunit. AB - We describe a novel genetic and kinetic defect in a slow-channel congenital myasthenic syndrome. The severely disabled propositus has advanced endplate myopathy, prolonged and biexponentially decaying endplate currents, and prolonged acetylcholine receptor (AChR) channel openings. Genetic analysis reveals the heterozygous mutation alphaV249F in the propositus and mosaicism for alphaV249F in the asymptomatic father. Unlike mutations described previously in the M2 transmembrane domain, alphaV249F is located N-terminal to the conserved leucines and is not predicted to face the channel lumen. Expression of the alphaV249F AChR in HEK fibroblasts demonstrates increased channel openings in the absence of ACh, prolonged openings in its presence, enhanced steady-state desensitization, and nanomolar rather than micromolar affinity of one of the two binding sites in the resting activatable state. Thus, neuromuscular transmission is compromised because cationic overloading leads to degenerating junctional folds and loss of AChR, because an increased fraction of AChR is desensitized in the resting state, and because physiological rates of stimulation elicit additional desensitization and depolarization block of transmission. PMID- 9221764 TI - Assembly and regulation of the CD40 receptor complex in human B cells. AB - CD40 is a member of the tumor necrosis factor (TNF) receptor superfamily. Studies with human B cells show that the binding of CD154 (gp39, CD40L) to CD40 recruits TNF receptor- associated factor 2 (TRAF2) and TRAF3 to the receptor complex, induces the downregulation of the nonreceptor-associated TRAFs in the cell and induces an increased expression of Fas on the cell surface. Combined signaling through the interluekin 4 receptor and CD40 induces an increased expression of Fas with a commensurate increase in the level of TRAF2, but not TRAF3, that is recruited to the receptor complex. In contrast, engagement of the membrane immunoglobulin and CD40 limits Fas upregulation and reduces the recruitment of TRAF2, relative to TRAF3, to the CD40 receptor complex. These studies show that the TRAF composition of the CD40 receptor complex can be altered by signals that influence B cell differentiation. PMID- 9221766 TI - Quantal neurotransmitter secretion rate exhibits fractal behavior. AB - The rate of exocytic events from both neurons and non-neuronal cells exhibits fluctuations consistent with fractal (self-similar) behavior in time, as evidenced by a number of statistical measures. We explicitly demonstrate this for neurotransmitter secretion at Xenopus neuromuscular junctions and for rat hippocampal synapses in culture; the exocytosis of exogenously supplied neurotransmitter from cultured Xenopus myocytes and from rat fibroblasts behaves similarly. The magnitude of the fluctuations of the rate of exocytic events about the mean decreases slowly as the rate is computed over longer and longer time periods, the periodogram decreases in power-law manner with frequency, and the Allan factor (relative variance of the number of exocytic events) increases as a power-law function of the counting time. These features are hallmarks of self similar behavior. Their description requires models that exhibit long-range correlation (memory) in event occurrences. We have developed a physiologically plausible model that accords with all of the statistical measures that we have examined. The appearance of fractal behavior at synapses, as well as in systems comprising collections of synapses, indicates that such behavior is ubiquitous in neural signaling. PMID- 9221767 TI - Neurotoxicity of the 22 kDa thrombin-cleavage fragment of apolipoprotein E and related synthetic peptides is receptor-mediated. AB - Potent neurotoxicity is associated with both apolipoprotein E (apoE)-related synthetic peptides and the 22 kDa N-terminal thrombin-cleavage fragment of apoE. Furthermore, the E4 isoform of the 22 kDa fragment is significantly more toxic than the same fragment derived from the E3 isoform, suggesting the possibility of a direct role of apoE-associated neurotoxicity in the pathophysiology of Alzheimer's disease. In the present study, the potential role of cell surface receptors in mediating neurotoxicity was assessed by using a variety of agents that should block the heparin-binding and receptor-binding activity of apoE. Effective inhibitors of neurotoxicity of both the apoE peptides and the apoE fragment include heparin, heparan sulfate, sodium chlorate and heparinase, the low-density lipoprotein (LDL) receptor-related protein receptor-associated protein, and a polyclonal anti-LDL receptor-related protein antibody. These results suggest that the neurotoxicity of the 22 kDa thrombin cleavage fragment of apoE and related peptides is receptor-mediated, and that the most likely candidate receptor is a heparan sulfate proteoglycan-LDL receptor-related protein complex. PMID- 9221768 TI - Characterization of guanylate kinase-associated protein, a postsynaptic density protein at excitatory synapses that interacts directly with postsynaptic density 95/synapse-associated protein 90. AB - The structure of central synapses is poorly understood at the molecular level. A recent advance came with the identification of the postsynaptic density-95 (PSD 95)/synapse-associated protein 90 family of proteins as important mediators of the synaptic clustering of certain classes of ion channels. By yeast two-hybrid screening, a novel protein termed guanylate kinase-associated protein (GKAP) has been isolated that binds to the GK-like domain of PSD-95 (). Here we present a detailed characterization of GKAP expression in the rat brain and report the cloning of a novel GKAP splice variant. By Northern blot, GKAP mRNAs (4, 6.5, and 8 kB) are expressed predominantly in the rat brain. By in situ hybridization, GKAP is expressed widely in neurons of cortex and hippocampus and in the Purkinje and granule cells of the cerebellum. On brain immunoblots, two prominent bands of 95 and 130 kDa are detected that correspond to products of short and long N terminal splice variants of GKAP. Two independent GKAP antibodies label somatodendritic puncta in neocortical and hippocampal neurons in a pattern consistent with synaptic elements. Immunogold electron microscopy reveals GKAP to be predominantly postsynaptic and present at asymmetric synapses and in dendritic spines. The distribution of GKAP immunogold particles is uniform in the lateral plane of the PSD but peaks in the perpendicular axis approximately 20 nm from the postsynaptic membrane. In cultured hippocampal neurons GKAP immunoreactive puncta colocalize with the AMPA receptor subunit Glu receptor 1 but not with the GABAA receptor subunits beta2 and beta3. Thus GKAP is a widely expressed neuronal protein localized specifically in the PSD of glutamatergic synapses, consistent with its direct interaction with PSD-95 family proteins. PMID- 9221769 TI - Dopamine depresses excitatory and inhibitory synaptic transmission by distinct mechanisms in the nucleus accumbens. AB - The release of dopamine (DA) in the nucleus accumbens (NAc) is thought to be critical for mediating natural rewards as well as for the reinforcing actions of drugs of abuse. DA and amphetamine depress both excitatory and inhibitory synaptic transmission in the NAc by a presynaptic D1-like DA receptor. However, the mechanisms of depression of excitatory and inhibitory synaptic transmission appear to be different. DA depressed the frequency of spontaneous miniature EPSCs, but the frequency of miniature IPSCs was depressed only when spontaneous release was made dependent on Ca2+ influx through voltage-dependent Ca2+ channels. Furthermore, the K+ channel blocker Ba2+ attenuated the effects of DA on evoked IPSPs, but not on EPSPs. Thus, DA appears to depress inhibitory synaptic transmission in the NAc by reducing Ca2+ influx into the presynaptic terminal, but depresses excitatory transmission by a distinct mechanism that is independent of the entry of Ca2+. PMID- 9221770 TI - High-affinity zinc inhibition of NMDA NR1-NR2A receptors. AB - Micromolar concentrations of extracellular Zn2+ are known to antagonize native NMDA receptors via a dual mechanism involving both a voltage-independent and a voltage-dependent inhibition. We have tried to evaluate the relative importance of these two effects and their subunit specificity on recombinant NMDA receptors expressed in HEK 293 cells and Xenopus oocytes. The comparison of NR1a-NR2A and NR1a-NR2B receptors shows that the voltage-dependent inhibition is similar in both types of receptors but that the voltage-independent inhibition occurs at much lower Zn2+ concentrations in NR1a-NR2A receptors (IC50 in the nanomolar range) than in NR1a-NR2B receptors (IC50 in the micromolar range). The high affinity of the effect observed with NR1a-NR2A receptors was found to be attributable mostly to the slow dissociation of Zn2+ from its binding site. By analyzing the effects of Zn2+ on varied combinations of NR1 (NR1a or NR1b) and NR2 (NR2A, NR2B, NR2C), we show that both the NR1 and the NR2 subunits contribute to the voltage-independent Zn2+ inhibition. We have observed further that under control conditions, i.e., in zero nominal Zn2+ solutions, the addition of low concentrations of heavy metal chelators markedly potentiates the responses of NR1a-NR2A receptors, but not of NR1a-NR2B receptors. This result suggests that traces of a heavy metal (probably Zn2+) contaminate standard solutions and tonically inhibit NR1a-NR2A receptors. Chelation of a contaminant metal also could account for the rapid NR2A subunit-specific potentiations produced by reducing compounds like DTT or glutathione. PMID- 9221771 TI - Selective destruction of stable microtubules and axons by inhibitors of protein serine/threonine phosphatases in cultured human neurons. AB - Paired helical filaments (PHFs) in the neurofibrillary tangles (NFTs) in Alzheimer's disease (AD) brains are composed of highly phosphorylated isoforms of tau (PHFtau) that fail to bind microtubules (MTs), and the levels of MT-binding competent tau are decreased in AD brains with abundant PHFtau. Because this loss of MT binding could compromise the viability of tangle-bearing AD neurons by destabilizing MTs, we asked whether these events could be initiated by inhibiting protein phosphatase 1 (PP1) and PP2A in cultured human neurons (NT2N cells) using okadaic acid (OK) and calyculin-A (CL-A). The treatment of NT2N cells with OK and CL-A increased tau phosphorylation, decreased the binding of tau to MTs, and selectively depolymerized the more stable detyrosinated MTs but not the more labile tyrosinated MTs. Significantly, this led to the rapid degeneration of axons, which are enriched in the more stable detyrosinated MTs, and PP2A was implicated in the initiation of this cascade of events because PP2A but not PP1 was closely associated with MTs in the NT2N cells. These studies imply that inactivation of PP2A in vulnerable neurons of the AD brain may play a mechanistic role in the conversion of normal tau into PHFtau, in the depolymerization of stable MTs, and in the degeneration of axons emanating from tangle-bearing neurons. PMID- 9221772 TI - Differential autoreceptor control of somatodendritic and axon terminal dopamine release in substantia nigra, ventral tegmental area, and striatum. AB - Dopamine (DA) is released from somatodendritic sites of neurons in the substantia nigra pars compacta (SNc) and ventral tegmental area (VTA), where it has neuromodulatory effects. The aim of this study was to evaluate the role of D2 autoreceptor inhibition in the regulation of this somatodendritic release in each region. Fast cyclic voltammetry at carbon fiber microelectrodes was used to measure electrically evoked DA release in vitro. Furthermore, we compared D2 regulation of somatodendritic release with the more familiar axon terminal release in caudate putamen (CPu) and nucleus accumbens (NAc). Evoked DA release was TTX-sensitive at all sites. There was significant D2 autoinhibition of DA release in SNc; however, this mechanism was two- to threefold less powerful, as compared with axon terminal release in CPu. In contrast to SNc, somatodendritic release in VTA was not under significant D2 receptor control, whereas release in the respective axon terminal region (NAc) was controlled strongly by autoinhibition. Thus, these data indicate that, first, autoinhibition via D2 receptors consistently plays a less significant role in the control of somatodendritic than axon terminal DA release, and, second, even at the level of somatodendrites themselves, D2 autoinhibition displays marked regional variation. In the light of previous data indicating that DA uptake processes are also less active in somatodendritic than in terminal regions, these results are interpreted as indicating that DA transmission is regulated differently in somatodendritic zones, as compared with axon terminals, and thus may have different functional consequences. PMID- 9221773 TI - Influence of subunit composition on desensitization of neuronal acetylcholine receptors at low concentrations of nicotine. AB - The influence of alpha and beta subunits on the properties of nicotine-induced activation and desensitization of neuronal nicotinic acetylcholine receptors (nAChRs) expressed in Xenopus oocytes was examined. Receptors containing alpha4 subunits were more sensitive to activation by nicotine than alpha3-containing receptors. At low concentrations of nicotine, nAChRs containing beta2 subunits reached near-maximal desensitization more rapidly than beta4-containing receptors. The concentration of nicotine producing half-maximal desensitization was influenced by the particular alpha subunit expressed; similar to results for activation, alpha4-containing receptors were more sensitive to desensitizing levels of nicotine than alpha3-containing receptors. The alpha subunit also influenced the rate of recovery from desensitization; this rate was approximately inversely proportional to the apparent nicotine affinity for the desensitized state. The homomeric alpha7 receptor showed the lowest sensitivity to nicotine for both activation and desensitization; alpha7 nAChRs also demonstrated the fastest desensitization kinetics. These subunit-dependent properties remained in the presence of external calcium, although subtle, receptor subtype-specific effects on both the apparent affinities for activation and desensitization and the desensitization kinetics were noted. These data imply that the subunit composition of various nAChRs determines the degree to which receptors are desensitized and/or activated by tobacco-related levels of nicotine. The subtype specific balance between receptor activation and desensitization should be considered important when the cellular and behavioral actions of nicotine are interpreted. PMID- 9221774 TI - A novel allosteric potentiator of AMPA receptors: 4--2 (phenylsulfonylamino)ethylthio--2,6-difluoro-phenoxyaceta mide. AB - We report that a novel sulfonylamino compound, 4-[2 (phenylsulfonylamino)ethylthio]-2,6-difluoro-phenoxyacetam ide (PEPA), selectively potentiates glutamate receptors of the AMPA subtype. PEPA (1-200 microM) dose dependently potentiated glutamate-evoked currents in Xenopus oocytes expressing AMPA (GluRA-GluRD), but not kainate (GluR6 and GluR6+KA2) or NMDA (zeta1 + epsilon1-epsilon4), receptor subunits. PEPA was effective at micromolar concentrations and, in contrast to the action of cyclothiazide, preferentially modulated AMPA receptor flop isoforms. At 200 microM, PEPA potentiated glutamate responses by 50-fold in oocytes expressing GluRCflop (EC50 approximately 50 microM) versus only threefold for GluRCflip; a similar preference for flop isoforms was observed for other AMPA receptor subunits. Dose-response analysis for GluRCflop revealed that 100 microM PEPA produced a sevenfold increase in AMPA receptor affinity for glutamate. PEPA produced considerably weaker potentiation of kainate-evoked than glutamate-evoked currents, suggesting modulation of the process of receptor desensitization. In human embryonic kidney 293 cells transfected with AMPA receptor subunits, PEPA either abolished or markedly slowed the rate of onset of desensitization and potentiated steady-state equilibrium currents evoked by glutamate with subunit (GluRC >/= GluRD > GluRA) and splice variant (flop > flip) selectivity similar to that observed in oocytes. Our results show that PEPA is a novel, flop-preferring allosteric modulator of AMPA receptor desensitization at least 100 times more potent than aniracetam. PMID- 9221775 TI - Ca2+- and voltage-dependent inactivation of Ca2+ channels in nerve terminals of the neurohypophysis. AB - Ca2+ channel inactivation was investigated in neurohypophysial nerve terminals by using patch-clamp techniques. The contribution of intracellular Ca2+ to inactivation was evaluated by replacing Ca2+ with Ba2+ or by including BAPTA in the internal recording solution. Ca2+ channel inactivation during depolarizing pulses was primarily voltage-dependent. A contribution of intracellular Ca2+ was revealed by comparing steady-state inactivation of Ca2+ channels with Ca2+ current and with intracellular [Ca2+]. However, this contribution was small compared to that of voltage. In contrast to voltage-gated Ca2+ channels in other preparations, in the neurohypophysis Ba2+ substitution or intracellular BAPTA increased the speed of inactivation while reducing the steady-state level of inactivation. Ca2+ channel recovery from inactivation was studied by using a paired-pulse protocol. The rate of Ca2+ channel recovery from inactivation at negative potentials was increased dramatically by Ba2+ substitution or intracellular BAPTA, indicating that intracellular Ca2+ inhibits recovery. Stimulation with trains of brief pulses designed to mimic physiological bursts of electrical activity showed that Ca2+ channel inactivation was much greater with 20 Hz trains than with 14 Hz trains. Inactivation induced by 20 Hz trains was reduced by intracellular BAPTA, suggesting an important role for Ca2+-dependent inactivation during physiologically relevant forms of electrical activity. Inhibitors of calmodulin and calcineurin had no effect on Ca2+ channel inactivation, arguing against a mechanism of inactivation involving these Ca2+ dependent proteins. The inactivation behavior described here, in which voltage effects on Ca2+ channel inactivation predominate at positive potentials and Ca2+ effects predominate at negative potentials, may be relevant to the regulation of neuropeptide release. PMID- 9221776 TI - Muscarinic modulation of spike backpropagation in the apical dendrites of hippocampal CA1 pyramidal neurons. AB - In pyramidal neurons from the CA1 region of the rat hippocampus, Na+-dependent action potentials backpropagate over the dendrites in an activity-dependent manner. Consequently, later spikes in a train have smaller amplitudes when recorded in the apical arbors. We studied the effect of the cholinergic agonist carbachol (CCh) on this pattern of activity when spikes were evoked synaptically or antidromically in the transverse slice preparation. Concentrations as low as 1 microM were effective in reversing the modulation, making the amplitude of all spikes in a train equal and independent of the frequency of spike firing. CCh did not change the propagation of the first spike in a train. These effects of CCh were blocked by 1 microM atropine, showing that only muscarinic receptors were involved. The effects of CCh on the pattern of spike propagation were observed in the proximal and middle dendrites, but recordings in the distal dendrites (>300 micron from the soma) showed that CCh did not boost the amplitude in this region. Intracellular BAPTA (10 mM) or EGTA (10 mM) had no effect on activity-dependent backpropagation but blocked the effect of CCh. Backpropagating spikes caused increases in [Ca2+]i at all dendritic locations. In the middle and distal dendrites these increases normally peaked at the time of the first few large action potentials. In association with the enhancement of spike backpropagation, CCh increased the amplitude and duration of the train-evoked [Ca2+]i changes. These effects of CCh on dendritic spike potentials and associated [Ca2+]i changes may be important in modulating synaptic integration and plasticity in these neurons. PMID- 9221777 TI - Requirement for tyrosine phosphatase during serotonergic neuromodulation by protein kinase C. AB - Tyrosine kinases and phosphatases are abundant in the nervous system, where they signal cellular differentiation, mediate the responses to growth factors, and direct neurite outgrowth during development. Tyrosine phosphorylation can also alter ion channel activity, but its physiological significance remains unclear. In an identified leech mechanosensory neuron, the ubiquitous neuromodulator serotonin increases the activity of a cation channel by activating protein kinase C (PKC), resulting in membrane depolarization and modulation of the receptive field properties. We observed that the effects on isolated neurons and channels were blocked by inhibiting tyrosine phosphatases. Serotonergic stimulation of PKC thus activates a tyrosine phosphatase activity associated with the channels, which reverses their constitutive inhibition by tyrosine phosphorylation, representing a novel form of neuromodulation. PMID- 9221778 TI - Direct recording of nicotinic responses in presynaptic nerve terminals. AB - Nicotinic acetylcholine receptors are widely expressed in the nervous system, but their functions remain poorly understood. One attractive hypothesis is that the receptors act presynaptically to modulate synaptic transmission. We provide a direct demonstration of presynaptic nicotinic receptors in situ by using whole cell patch-clamp techniques to record currents in large presynaptic calyces that midbrain neurons form on ciliary neurons. Bath application of nicotine induced inward currents in the calyces capable of generating action potentials that overrode the limited space clamp achievable. The inward currents reversed near 0 mV and showed inward rectification common for neuronal nicotinic receptors. Tetrodotoxin (TTX) blocked the action potentials but not the inward currents. alpha-Bungarotoxin blocked both, consistent with the presynaptic receptors containing alpha7 subunits. Recording from the postsynaptic ciliary neurons during nicotine exposure revealed EPSCs that TTX blocked, presumably by blocking presynaptic action potentials. The postsynaptic cells also displayed bimodal inward currents caused by their own nicotinic receptors; the bimodal currents were not blocked by TTX but were blocked partially by alpha-bungarotoxin and completely by D-tubocurarine. Dye-filling with Lucifer yellow from the recording pipette confirmed the identity of patched structures and showed no dye transfer between calyx and ciliary neuron. When calyces or ciliary neurons were labeled en mass with neurobiotin and biocytin through nerve roots, dye transfer was rarely observed. Thus, electrical synapses were infrequent and unlikely to influence calyx responses. Immunochemical analysis of preganglionic nerve extracts identified receptors that bind alpha-bungarotoxin and contain alpha7 subunits. The results unambiguously document the existence of functional presynaptic nicotinic receptors. PMID- 9221779 TI - Microtubule transport from the cell body into the axons of growing neurons. AB - The present studies test the hypothesis that microtubules (MTs) are transported from the cell body into the axons of growing neurons. Dissociated sympathetic neurons were cultured using conditions that allow us to control the initiation of axon outgrowth. Neurons were injected with biotin-labeled tubulin (Bt-tub) and then stimulated to extend axons. The newly formed axons were then examined using immunofluorescence procedures for MTs with or without Bt-tub. Because the Bt-tub is fully assembly competent, all MTs that assemble after injection will contain Bt-tub. However, MTs that exist in the neuron at the time of injection and persist during the subsequent incubation will not contain Bt-tub. Because the neurons were injected before extending axons, MTs without Bt-tub are initially localized to the cell body. We specifically determined whether these MTs appeared in the newly formed axon. Such a result can only be explained by the transport of these MTs from their initial location in the cell body into the axon. The newly formed axons of many neurons contained MTs both with and without Bt-tub. MTs without Bt-tub were detected all along the axon and in some neurons represented a substantial portion of the total polymer in the proximal and middle regions of the axon. These results show that MTs are transported from the cell body into growing axons and that this transport is robust, delivering MTs to all regions of the newly formed axon. PMID- 9221780 TI - Epidermal growth factor and fibroblast growth factor-2 have different effects on neural progenitors in the adult rat brain. AB - Neurons and glia are generated throughout adulthood from proliferating cells in two regions of the rat brain, the subventricular zone (SVZ) and the hippocampus. This study shows that exogenous basic fibroblast growth factor (FGF-2) and epidermal growth factor (EGF) have differential and site-specific effects on progenitor cells in vivo. Both growth factors expanded the SVZ progenitor population after 2 weeks of intracerebroventricular administration, but only FGF 2 induced an increase in the number of newborn cells, most prominently neurons, in the olfactory bulb, the normal destination for neuronal progenitors migrating from the SVZ. EGF, on the other hand, reduced the total number of newborn neurons reaching the olfactory bulb and substantially enhanced the generation of astrocytes in the olfactory bulb. Moreover, EGF increased the number of newborn cells in the striatum either by migration of SVZ cells or by stimulation of local progenitor cells. No evidence of neuronal differentiation of newborn striatal cells was found by three-dimensional confocal analysis, although many of these newborn cells were associated closely with striatal neurons. The proliferation of hippocampal progenitors was not affected by either growth factor. However, EGF increased the number of newborn glia and reduced the number of newborn neurons, similar to the effects seen in the olfactory bulb. These findings may be useful for elucidating the in vivo role of growth factors in neurogenesis in the adult CNS and may aid development of neuronal replacement strategies after brain damage. PMID- 9221781 TI - OCAM: A new member of the neural cell adhesion molecule family related to zone-to zone projection of olfactory and vomeronasal axons. AB - Zone-to-zone projection of olfactory and vomeronasal sensory axons underlies the topographic and functional mapping of chemoreceptor expression zones of the sensory epithelia onto zonally arranged glomeruli in the main and accessory olfactory bulbs. Here we identified OCAM (R4B12 antigen), an axonal surface glycoprotein expressed by subsets of both olfactory and vomeronasal axons in a zone-specific manner. OCAM is a novel homophilic adhesion molecule belonging to the immunoglobulin superfamily with striking structural homology to neural cell adhesion molecule. In both the main and accessory olfactory systems, OCAM mRNA is expressed by sensory neurons in restricted chemoreceptor expression zones, and OCAM protein-expressing axons project to the glomeruli in the corresponding zones of the main and accessory bulbs. OCAM protein is expressed on subsets of growing sensory axons in explant cultures even in the absence of the target bulb. These results demonstrate a precisely coordinated zonal expression of chemoreceptors and OCAM and suggest that OCAM may play important roles in selective fasciculation and zone-to-zone projection of the primary olfactory axons. PMID- 9221782 TI - Caenorhabditis elegans levamisole resistance genes lev-1, unc-29, and unc-38 encode functional nicotinic acetylcholine receptor subunits. AB - We show that three of the eleven genes of the nematode Caenorhabditis elegans that mediate resistance to the nematocide levamisole and to other cholinergic agonists encode nicotinic acetylcholine receptor (nAChR) subunits. unc-38 encodes an alpha subunit while lev-1 and unc-29 encode non-alpha subunits. The nematode nAChR subunits show conservation of many mammalian nAChR sequence features, implying an ancient evolutionary origin of nAChR proteins. Expression in Xenopus oocytes of combinations of these subunits that include the unc-38 alpha subunit results in levamisole-induced currents that are suppressed by the nAChR antagonists mecamylamine, neosurugatoxin, and d-tubocurarine but not alpha bungarotoxin. The mutant phenotypes reveal that unc-38 and unc-29 subunits are necessary for nAChR function, whereas the lev-1 subunit is not. An UNC-29-GFP fusion shows that UNC-29 is expressed in body and head muscles. Two dominant mutations of lev-1 result in a single amino acid substitution or addition in or near transmembrane domain 2, a region important to ion channel conductance and desensitization. The identification of viable nAChR mutants in C. elegans provides an advantageous system in which receptor expression and synaptic targeting can be manipulated and studied in vivo. PMID- 9221783 TI - Quantitative ultrastructural analysis of hippocampal excitatory synapses. AB - From three-dimensional reconstructions of CA1 excitatory synapses in the rodent hippocampus and in culture, we have estimated statistical distributions of active zone and postsynaptic density (PSD) sizes (average area approximately 0.04 micron2), the number of active zones per bouton (usually one), the number of docked vesicles per active zone (approximately 10), and the total number of vesicles per bouton (approximately 200), and we have determined relationships between these quantities, all of which vary from synapse to synapse but are highly correlated. These measurements have been related to synaptic physiology. In particular, we propose that the distribution of active zone areas can account for the distribution of synaptic release probabilities and that each active zone constitutes a release site as identified in the standard quantal theory attributable to Katz (1969). PMID- 9221784 TI - Cortistatin is expressed in a distinct subset of cortical interneurons. AB - Cortistatin is a presumptive neuropeptide that shares 11 of its 14 amino acids with somatostatin. In contrast to somatostatin, administration of cortistatin into the rat brain ventricles specifically enhances slow wave sleep, apparently by antagonizing the effects of acetylcholine on cortical excitability. Here we show that preprocortistatin mRNA is expressed in a subset of GABAergic cells in the cortex and hippocampus that partially overlap with those containing somatostatin. A significant percentage of cortistatin-positive neurons is also positive for parvalbumin. In contrast, no colocalization was found between cortistatin and calretinin, cholecystokinin, or vasoactive intestinal peptide. During development there is a transient increase in cortistatin-expressing cells in the second postnatal week in all cortical areas and in the dentate gyrus. A transient expression of preprocortistatin mRNA in the hilar region at P16 is paralleled by electrophysiological changes in dentate granule cells. Together, these observations suggest mechanisms by which cortistatin may regulate cortical activity. PMID- 9221785 TI - Mammalian homolog of Drosophila retinal degeneration B rescues the mutant fly phenotype. AB - Mutations in the Drosophila rdgB gene, which encodes a transmembrane phosphatidylinositol transfer protein (PITP), cause a light-enhanced retinal degeneration. Cloning of mammalian rdgB orthologs (mrdgB) reveal predicted proteins that are 39% identical to rdgB, with highest homology in the N-terminal PITP domain (62%) and in a region near the C terminus (65%). The human mrdgB gene spans approximately 12 kb and maps to 11q13.1, a locus where several retinal diseases have also been mapped. Murine mrdgB maps to a syntenic region on the proximal region of chromosome 19. MrdgB is specifically expressed in the retina and brain. In the retina, MrdgB protein is localized to photoreceptor inner segments and the outer and inner plexiform layers. Expression of murine mrdgB in mutant flies fully rescues both the rdgB-dependent retinal degeneration and abnormal electroretinogram. These results suggest the existence of similarities between the invertebrate and mammalian retina that were not previously appreciated and also identify mrdgB as a candidate gene for retinal diseases that map to 11q13.1. PMID- 9221786 TI - Sonic hedgehog promotes the survival of specific CNS neuron populations and protects these cells from toxic insult In vitro. AB - Sonic hedgehog (Shh), an axis-determining secreted protein, is expressed during early vertebrate embryogenesis in the notochord and ventral neural tube. In this site it plays a role in the phenotypic specification of ventral neurons along the length of the CNS. For example, Shh induces the differentiation of motor neurons in the spinal cord and dopaminergic neurons in the midbrain. Shh expression, however, persists beyond this induction period, and we have asked whether the protein shows novel activities beyond phenotype specification. Using cultures derived from embryonic day 14.5 (E14. 5) rat ventral mesencephalon, we show that Shh is also trophic for dopaminergic neurons. Interestingly, Shh not only promotes dopaminergic neuron survival, but also promotes the survival of midbrain GABA-immunoreactive (GABA-ir) neurons. In cultures derived from the E15-16 striatum, Shh promotes the survival of GABA-ir interneurons to the exclusion of any other cell type. Cultures derived from E15-16 ventral spinal cord reveal that Shh is again trophic for interneurons, many of which are GABA-ir and some of which express the Lim-1/2 nuclear marker, but it does not appear to support motorneuron survival. Shh does not support the survival of sympathetic or dorsal root ganglion neurons. Finally, using the midbrain cultures, we show that in the presence of MPP+, a highly specific neurotoxin, Shh prevents dopaminergic neuron death that normally would have occurred. Thus Shh may have therapeutic value as a protective agent in neurodegenerative disease. PMID- 9221787 TI - Path integration and cognitive mapping in a continuous attractor neural network model. AB - A minimal synaptic architecture is proposed for how the brain might perform path integration by computing the next internal representation of self-location from the current representation and from the perceived velocity of motion. In the model, a place-cell assembly called a "chart" contains a two-dimensional attractor set called an "attractor map" that can be used to represent coordinates in any arbitrary environment, once associative binding has occurred between chart locations and sensory inputs. In hippocampus, there are different spatial relations among place fields in different environments and behavioral contexts. Thus, the same units may participate in many charts, and it is shown that the number of uncorrelated charts that can be encoded in the same recurrent network is potentially quite large. According to this theory, the firing of a given place cell is primarily a cooperative effect of the activity of its neighbors on the currently active chart. Therefore, it is not particularly useful to think of place cells as encoding any particular external object or event. Because of its recurrent connections, hippocampal field CA3 is proposed as a possible location for this "multichart" architecture; however, other implementations in anatomy would not invalidate the main concepts. The model is implemented numerically both as a network of integrate-and-fire units and as a "macroscopic" (with respect to the space of states) description of the system, based on a continuous approximation defined by a system of stochastic differential equations. It provides an explanation for a number of hitherto perplexing observations on hippocampal place fields, including doubling, vanishing, reshaping in distorted environments, acquiring directionality in a two-goal shuttling task, rapid formation in a novel environment, and slow rotation after disorientation. The model makes several new predictions about the expected properties of hippocampal place cells and other cells of the proposed network. PMID- 9221788 TI - Noxious cutaneous thermal stimuli induce a graded release of endogenous substance P in the spinal cord: imaging peptide action in vivo. AB - Dorsal root ganglia (DRG) neurons synthesize and transport substance P (SP) to the spinal cord where it is released in response to intense noxious somatosensory stimuli. We have shown previously that SP release in vivo causes a rapid and reversible internalization of SP receptors (SPRs) in dorsal horn neurons, which may provide a pharmacologically specific image of neurons activated by SP. Here, we report that noxious heat (43 degrees, 48 degrees, and 55 degrees C) and cold (10 degrees, 0 degrees, -10 degrees, and -20 degrees C) stimuli, but not innocuous warm (38 degrees C) and cold (20 degrees C) stimuli, applied to the hindpaw of anesthetized rats induce SPR internalization in spinal cord neurons that is graded with respect to the intensity of the thermal stimulus. Thus, with increasing stimulus intensities, both the total number of SPR+ lamina I neurons showing SPR internalization and the number of internalized SPR+ endosomes within each SPR immunoreactive neuron showed a significant increase. These data suggest that thermal stimuli induce a graded release of SP from primary afferent terminals and that agonist dependent receptor endocytosis provides evidence of a spatially and pharmacologically unique "neurochemical signature" after specific somatosensory stimuli. PMID- 9221789 TI - AMPA receptor facilitation accelerates fear learning without altering the level of conditioned fear acquired. AB - Rats treated with the AMPA receptor-facilitating drug 1-(quinoxolin-6 ylcarbonyl)piperidine (BDP-12) during training acquired fear conditioning to a tone faster than vehicle-treated controls. The effect on acquisition was dependent on the dose given. BDP-12-treated rats and vehicle-treated controls reached the same level of conditioned fear and extinguished at the same rate. The dissociation of learning rate from these other normally covariant measures suggests that the drug had a specific and isolated effect on acquisition. Controls for drug effects on stimulus sensitivity, locomotor activity, generalized fearfulness, and other performance factors support this interpretation. The known action of BDP-12 on receptor dynamics suggests that its effect on acquisition may be attributed to specific modulation of an AMPA and NMDA receptor-dependent plasticity mechanism. The finding that the drug accelerates acquisition but does not affect the level of conditioned fear acquired parallels the effect of the drug on long-term potentiation (LTP) (increasing the rate but not the ceiling of potentiation) and suggests that common mechanisms may underlie fear conditioning and LTP. PMID- 9221790 TI - Contributions of voltage-gated Ca2+ channels in the proximal versus distal dendrites to synaptic integration in prefrontal cortical neurons. AB - The electrogenesis of synaptically activated dendritic Ca2+-mediated potentials, which may contribute to synaptic signal integration in pyramidal cells, was examined in rat layers V-VI prefrontal cortical (PFC) neurons in vitro. Intrasomatically recorded suprathreshold synaptic responses evoked by stimulation of the distal dendrites were attenuated by focal Cd2+ application to the proximal apical dendritic stem (100-200 micron from soma), but not to the apical dendritic tuft (>500 micron from soma). With use of intracellular QX-314 and Cs+ to block Na+ and K+ currents, intrasomatic recordings revealed that the Cd2+-induced attenuation of synaptic responses was attributable to the blockade of a dendritic Ca2+-mediated "hump" potential and high-threshold Ca2+ spike activated by NMDA EPSPs. The hump potential was not blocked by bath application of Ni2+ (100 microM) but was blocked by focal application of Cd2+ to the proximal but not distal apical dendrites, suggesting that it was generated by Ca2+ channels located in the proximal dendrites. Direct patch-clamp recordings made from the distal apical tuft of layers V-VI PFC neurons revealed that layers I-II synaptic stimulation or intradendritic depolarizing current pulses evoked tetrodotoxin- and QX-314-sensitive Na+ spikes. Unlike in the stem of the apical dendrite, Ca2+ spikes were not easily evoked in the distal apical tuft when Na+ channels were blocked. When triggered, the Cd2+-sensitive Ca2+ spikes in the dendritic tuft were nonregenerative and had very high activation thresholds (approximately +10 mV). These results suggested that the high voltage-activated Ca2+ potentials that amplify distal EPSPs are primarily generated in the proximal stem of the apical dendrite and not within the fine dendritic branches of the apical tuft of layers V-VI PFC neurons. PMID- 9221791 TI - Mice lacking the TNF 55 kDa receptor fail to sleep more after TNFalpha treatment. AB - Tumor necrosis factor (TNF) is a well characterized sleep-regulatory substance. To study receptor mechanisms for the sleep-promoting effects of TNF, sleep patterns were determined in control and TNF 55 kDa receptor knock-out (TNFR-KO) mice with a B6 x 129 background after intraperitoneal injections of saline or murine TNFalpha. The TNFR-KO mice had significantly less baseline sleep than the controls. TNFalpha dose-dependently increased non-rapid eye movement sleep (NREMS) in the controls but did not influence sleep in TNFR-KO mice. Although TNFR-KO mice failed to respond to TNFalpha, they had an increase in NREMS and a decrease in rapid eye movement sleep after interleukin-1beta treatment. These results indicate that TNFalpha affects sleep via the 55 kDa receptor and provide further evidence that TNFalpha is involved in physiological sleep regulation. Current results also extend the list of species to mice in which TNFalpha and interleukin-1beta are somnogenic. PMID- 9221792 TI - Muscle response to changing neuronal input in the lobster (Panulirus interruptus) stomatogastric system: spike number- versus spike frequency-dependent domains. AB - We aimed to determine the neuronal parameters controlling the contraction of slowly contracting, non-twitch ("tonic") muscles driven by rhythmic neuronal activity. These muscles are almost completely absent in mammals but are common in lower vertebrates and invertebrates. Slow muscles are often believed to function primarily in tonic motor patterns. However, previous research and data presented here indicate that slow muscles are also driven by rhythmic neuronal inputs. In rapidly contracting "twitch" muscles, motor unit force is believed to be primarily determined by motor neuron spike frequency. What determines slow muscle output is less well understood. We present a simple model that suggests that when motor neuron burst duration is brief compared with muscle summation time, spike number, not spike frequency, determines slow muscle contraction amplitude. We present analyses that distinguish between spike number and spike frequency dependence in two slow muscles in the lobster stomatogastric system. Our analysis shows that, functionally, one muscle is spike number dependent, whereas the other is primarily spike frequency dependent. Thus, both of these parameters can determine slow muscle output. To predict the movements elicited by neuronal activity in preparations in which slow muscles are common, it may be necessary to determine spike number versus spike frequency dependence for each muscle. Spike number dependence couples motor neuron burst duration and spike frequency in that changing either parameter alone alters spike number (and hence muscle contraction amplitude). Neural networks innervating spike number-dependent muscles may therefore have specific properties to compensate for the complexity intrinsic to spike number coding. PMID- 9221793 TI - Prolonged and extrasynaptic excitatory action of dopamine mediated by D1 receptors in the rat striatum in vivo. AB - The spatiotemporal characteristics of the dopaminergic transmission mediated by D1 receptors were investigated in vivo. For this purpose dopamine (DA) release was evoked in the striatum of anesthetized rats by train electrical stimulations of the medial forebrain bundle (one to four pulses at 15 Hz), which mimicked the spontaneous activity of dopaminergic neurons. The resulting dopamine overflow was electrochemically monitored in real time in the extracellular space. This evoked DA release induced a delayed increase in discharge activity in a subpopulation of single striatal neurons. This excitation was attributable to stimulation of D1 receptors by released DA because it was abolished by acute 6-hydroxydopamine lesion and strongly reduced by the D1 antagonist SCH 23390. Striatal neurons exhibiting this delayed response were also strongly excited by intravenous administration of the D1 agonist SKF 82958. Whereas the DA overflow was closely time-correlated with stimulation, the excitatory response mediated by DA started 200 msec after release and lasted for up to 1 sec. Moreover, functional evidence presented here combined with previous morphological data show that D1 receptors are stimulated by DA diffusing up to 12 micron away from release sites in the extrasynaptic extracellular space. In conclusion, DA released by bursts of action potentials exerts, via D1 receptors, a delayed and prolonged excitatory influence on target neurons. This phasic transmission occurs outside synaptic clefts but still exhibits a high degree of spatial specificity. PMID- 9221794 TI - Lesions of the medial geniculate nuclei specifically block corticosterone release and induction of c-fos mRNA in the forebrain associated with audiogenic stress in rats. AB - Audiogenic stress is known to activate the hypothalamo-pituitary-adrenocortical (HPA) axis in rats. The goal of the present study was to determine whether the medial geniculate nuclei (including all auditory nuclei of the thalamus), which are obligatory relays in the transmission of auditory information to the forebrain, are critically involved in HPA activation by audiogenic stress. To this end, corticosterone levels and regional brain activity indexed by c-fos mRNA induction, elicited by 30 min of 105 dB white noise, were measured. Compared with unoperated and sham-operated rats, complete medial geniculate nuclei lesions blocked corticosterone release normally induced by loud noise. The effects of the lesions were specific to loud noise insofar as corticosterone release in response to restraint or ether stress was not reduced in lesioned rats. We have determined previously that audiogenic stress is associated with a specific regional pattern of c-fos mRNA induction. Rats sustaining complete medial geniculate lesions demonstrated a blockade of c-fos mRNA induction in several audiogenic stress responsive regions, also known to directly innervate medial parvocellular neurons of the paraventricular hypothalamic nucleus. Thus, in addition to blockade in the paraventricular hypothalamic nucleus, c-fos mRNA induction in the lesioned animals was abolished in the bed nucleus of the stria terminalis, especially its anterior medial and ventral aspects, the septohypothalamic nucleus, and the anteroventral preoptic area, compared with unoperated and sham-operated rats. Several additional regions in the lesioned rats failed to show reliable c-fos mRNA induction compared with naive rat controls. Nearly all other regions that showed reliable c-fos mRNA induction in the unoperated and sham-operated rats displayed either similar or slightly reduced levels in complete medial geniculate lesioned rats, suggesting that these regions are not part of a critical HPA activational circuit in response to audiogenic stress. On the basis of these results, putative circuits from the medial geniculate nuclei to the paraventricular nucleus of the hypothalamus involved in activation of the HPA axis by audiogenic stress are discussed. PMID- 9221795 TI - rGbeta1: a psychostimulant-regulated gene essential for establishing cocaine sensitization. AB - Repeated doses of cocaine or amphetamine lead to long-lasting behavioral manifestations that include enhanced responses termed sensitization. Although biochemical mechanisms that underlie these manifestations currently remain largely unknown, new protein synthesis has been implicated in several of these neuroadaptive processes. To seek candidate biochemical mechanisms for these drug induced neuroplastic behavioral responses, we have used an approach termed subtracted differential display (SDD) to identify genes whose expression is regulated by these psychostimulants. rGbeta1 is one of the SDD products that encodes a rat G-protein beta subunit. rGbeta1 expression is upregulated by cocaine or amphetamine treatments in neurons of the nucleus accumbens shell region, a major center for psychostimulant effects in locomotor control and behavioral reward. Antisense oligonucleotide treatments that attenuate rGbeta1 expression in regions including the nucleus accumbens abolish the development of behavioral sensitization when they are administrated during the repeated cocaine exposures that establish sensitization. These treatments fail to alter acute behavioral responses to cocaine, and they do not block the expression of cocaine sensitization when it is established before oligonucleotide administrations. Full, regulated rGbeta1 expression is a biochemical component essential to the establishment of a key consequence of repeated cocaine administrations, sensitization. PMID- 9221796 TI - Seasonal changes in testosterone, neural attributes of song control nuclei, and song structure in wild songbirds. AB - Seasonal changes in the neural attributes of brain nuclei that control song in songbirds are among the most pronounced examples of naturally occurring plasticity in the adult brain of any vertebrate. The behavioral correlates of this seasonal neural plasticity have not been well characterized, particularly in songbird species that lack adult song learning. To address this question, we investigated the relationship between seasonal changes in gonadal steroids, song nuclei, and song behavior in adult male song sparrows (Melospiza melodia). At four times of the year, we measured plasma concentrations of testosterone, neural attributes of song nuclei, and several aspects of song structure in wild song sparrows of a nonmigratory population. We found seasonal changes in the song nuclei that were temporally correlated with changes in testosterone concentrations and with changes in song stereotypy. Male song sparrows sang songs that were more variable in structure in the fall, when testosterone concentrations were low and song nuclei were small, than in the spring, when testosterone concentrations were higher and song nuclei were larger. Despite seasonal changes in the song nuclei, the song sparrows continued to sing the same number of different song types, indicating that changes in the song nuclei were not correlated with changes in song repertoire size. These results suggest that song stereotypy, but not repertoire size, is a potential behavioral correlate of seasonal plasticity in the avian song control system. PMID- 9221797 TI - Excitotoxic lesions of the amygdala fail to produce impairment in visual learning for auditory secondary reinforcement but interfere with reinforcer devaluation effects in rhesus monkeys. AB - Aspiration lesions of the amygdala were found previously to produce a severe impairment in visual discrimination learning for auditory secondary reinforcement in rhesus monkeys (Gaffan and Harrison, 1987). To determine whether excitotoxic amygdala lesions would also produce this effect, we trained four naive rhesus monkeys on the same task. The monkeys were required to learn 40 new visual discrimination problems per session in a situation in which visual choices were guided by an auditory secondary reinforcer that had been previously associated with food reward. Bilateral excitotoxic lesions of the amygdala had no effect on the rate of learning visual discrimination problems for auditory secondary reinforcement. We also tested the amygdalectomized monkeys on a reinforcer devaluation task and compared their performance with a group of three normal monkeys. The monkeys first learned to discriminate 60 pairs of objects, baited with two different food rewards. Each of the food rewards was then devalued by selective satiation in two separate experimental sessions. Normal controls tended to avoid displacing objects that covered the devalued food to a significantly greater degree than did the amygdalectomized monkeys, indicating that the excitotoxic amygdala damage interfered with reinforcer devaluation effects. Our results are consistent with the idea that the amygdala is necessary for learning the association between stimuli and the value of particular food rewards; however, the amygdala is not necessary for maintaining the value of secondary reinforcers, once they have been learned. PMID- 9221798 TI - Expression of pS2 protein in endometrial carcinomas: correlation with clinicopathologic features and sex steroid receptor status. AB - Using immunohistochemistry, we examined pS2 expression in 64 samples of endometrial carcinoma, 11 samples of endometrial hyperplasia and 15 samples of normal endometrium, and compared them with clinicopathological data, estrogen receptor (ER) expression and progesterone receptor (PR) expression. Of the 64 samples of endometrial carcinoma, 45 (70%) expressed the pS2 protein. The average age of the patients with pS2-positive carcinomas (54.8 +/- 8.6 years) was significantly lower than that of the patients with pS2-negative carcinomas, and all premenopausal patients were positive for the pS2 protein. Among histological types, pS2 expression was observed in 33 (92%) of the 36 G1 carcinomas, but in none of the 5 nonendometrioid carcinomas. Of the 48 ER-positive carcinomas, 43 (90%) were pS2-positive and 5 were pS2-negative. Of the 40 PR-positive carcinomas, 37 (93%) were positive for pS2. There were significant associations between pS2 expression and ER/PR expression (p < 0.001). Staining of the pS2 protein was also observed in the samples of normal endometrium. We found a progressive increase in immunoreactivity of pS2 protein from normal endometrium to endometrial hyperplasia and still more in well-differentiated carcinoma. All 11 cases of endometrial hyperplasia were strongly positive for pS2. Furthermore, patients with pS2-positive carcinomas had a better survival rate than those with pS2-negative carcinomas (p < 0.05). Our data suggest that pS2 expression is likely correlated with estrogen-related endometrial carcinoma and is possibly involved in early disease progression. PMID- 9221799 TI - Group II phospholipase A2 is increased in peritoneal and pleural effusions in patients with various types of cancer. AB - Serum levels of group II phospholipase A2 (PLA2) have been reported to be associated with stage of disease in cancer patients. These levels are also related to the malignant potential in tissues, and are an important prognostic factor. We radioimmunoassayed group II PLA2 levels in pleural and peritoneal effusions from patients with various cancers. We also investigated the production of group II PLA2 in cells in effusions from cancer patients by Northern blotting, immunocytochemistry and in situ hybridization. Immunoreactive group II PLA2 levels were significantly higher in effusions from 47 patients with various cancers, compared with those in sera and cirrhotic ascites. There was no significant correlation between group II PLA2 levels in effusions and those in sera. Group II PLA2 mRNA was expressed at a high level in cells from effusions, by Northern blot analysis, but not in those cells from blood. The localization of group II PLA2 protein and mRNA was intense in carcinoma cells and CD68-positive macrophages, determined by immunocytochemistry and in situ hybridization. In addition, IL-6 and IL-8 levels were significantly higher in effusions, in comparison with those in sera from patients, suggesting that cancer cells and macrophages produce group II PLA2 by IL-6. These group II PLA2 levels are apparently significantly increased in effusions, and the carcinoma cells and macrophages produce group II PLA2, as noted in effusions from patients with various cancers. PMID- 9221800 TI - Association of reduced cell adhesion regulator messenger RNA expression with tumor progression in human hepatocellular carcinoma. AB - The recently identified cell adhesion regulator (CAR) modulates the process of integrin-mediated cell adhesion. The CAR gene is located on 16q, a locus at which high levels of allelic losses have been demonstrated in advanced human hepatocellular carcinoma (HCC). We studied the possible involvement of the CAR gene in the progression of HCC. With this aim, we determined the expression of CAR mRNA in 30 cases of HCC. Matching pair samples of tumor and adjacent nontumoral liver were analyzed by semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR). The results were compared with the clinicopathological features of the patients. Every nontumoral liver tissue sample analyzed, expressed CAR mRNA. All tumor samples showed amounts of expression that were equal or lower, compared with those found in their matching controls. Thus, in 16 out of 30 cases (53.3%), CAR mRNA expression in tumor was diminished to less than one tenth of that observed in nontumoral tissue. This group of patients exhibited higher amounts of alpha-fetoprotein, and comprised tumors with poor histological differentiation (Edmondson-Steinert's grades III IV), higher rates of intrahepatic metastasis and recurrence within the first postoperative year (p < 0.05, respectively). Tumors exhibiting low levels of CAR mRNA were also found to be diagnosed at more advanced TNM stages (p < 0.01). We conclude that downregulation of CAR mRNA expression may play an essential role in the progression of HCC. PMID- 9221801 TI - Loss of expression of the p16INK4/CDKN2 gene in cutaneous malignant melanoma correlates with tumor cell proliferation and invasive stage. AB - The G1/S checkpoint of the cell cycle is regulated by p16, p53 and RB tumor suppressor genes. Loss of expression of the p16INK4 tumor suppressor protein, the product of the CDKN2 gene, has been associated with a wide variety of human malignancies. Mutations, loss of heterozygosity and deletions of the CDKN2 locus have been reported in sporadic and familial cutaneous malignant melanomas (CMM). To investigate the role of the alterations of p16 expression in melanoma, we evaluated by immunohistochemistry the p16 expression and cell proliferation in 79 primary CMM and 10 benign melanocytic nevi (BMN). Forty-six melanomas (58%) and all BMN were found to be p16 positive; 33 melanomas (42%) were considered p16 negative. The extent of invasion according to Clark was significantly higher in p16-negative tumors than in p16-positive tumors. Cell proliferation as expressed by the proportion of positive cells in Ki-67 immunostaining was found to be significantly higher in p16-negative tumors than in p16-positive tumors, although there was no significant difference in the mitotic index between p16-positive and p16-negative tumors. In p16-positive tumors, the number of Ki-67-positive cells correlated with the mitotic index; in p16-negative tumors, there was no correlation between these parameters. Our data suggest that loss of p16 expression is more common in advanced melanomas, and that G1/S checkpoint regulation is disrupted in p16-negative melanomas. Our results show that loss of p16 expression is a common event in primary melanomas, which further substantiates the role of p16 as a major tumor suppressor. PMID- 9221802 TI - Prognostic implications of different cell cycle analysis models of flow cytometric DNA histograms of 1,301 breast cancer patients: results from the Multicenter Morphometric Mammary Carcinoma Project (MMMCP). AB - Conflicting prognostic results with regard to DNA flow cytometric cell cycle variables have been reported for breast cancer patients. An important reason for this may be related to differences in the interpretation of DNA histograms. Several computer programs based on different cell cycle fitting models are available resulting in significant variations in percent S-phase and other cell cycle variables. Our present study evaluated the prognostic value of percent S phase cells obtained using 5 different cell cycle analysis models. Flow cytometric DNA histograms obtained from 1,301 fresh frozen breast cancer samples were interpreted with 5 different cell cycle analysis models using a commercially available computer program. Model 1 used the zero order S-phase calculation and "sliced nuclei" debris correction, model 2 added fixed G2/M- to G0/G1-phase ratio, and model 3 added correction for aggregates. Model 4 applied the first order S-phase calculation and sliced debris correction. Model 5 fixed the coefficients of variation CVs of the G0/G1- and G2/M-phases in addition to applying the sliced nuclei debris correction and zero order S-phase calculation. The different models yielded clearly different prognostic results. The average percent S-phase cells of the aggregate correction model (model 3) provided the best prognostic value in all cases for overall survival (OS) as well as disease free survival (DFS) (OS: p < 0.0001; DFS: p < 0.0001), in lymph node-positive cases (OS: p < 0.0001; DFS: p = 0.004) and in DNA-diploid subgroups (OS: p = 0.004; DFS: p = 0.001). For the lymph node negative and DNA-non-diploid subgroups, the percent S-phase of the second cell cycle reached slightly better prognostic significance than the average percent S-phase cells. In multivariate analysis, the average percent S-phase of the aggregate correction model had the best additional prognostic value to tumor size and lymph node status. In conclusion, different cell cycle analysis models yield clearly different prognostic results for invasive breast cancer patients. The most important prognostic percent S-phase variable was the average percent S-phase cells when aggregate correction was included in cell cycle analysis. PMID- 9221803 TI - Expression of a breast-cancer-associated protein (pS2) in human neuro-endocrine tumours. AB - pS2 protein expression has been demonstrated in a range of malignant tissues in an oestrogen-independent pathway. Recently, it has been demonstrated that pS2, in prostate cancer, is closely associated with neuro-endocrine differentiation. In the present study, we have analyzed, by immunohistochemistry along with microwave antigen retrieval, the expression of pS2 protein in a retrospective series of 236 human primary neuro-endocrine tumours and attempted to correlate this with the clinicopathologic features of patients and the presence of oestrogen receptor (ER). pS2 immunoreactivity was detected in 42% of small-cell lung carcinomas, 36% of lung carcinoids, 33% of phaeochromocytomas, 38% of carotid-body tumours, 31% of pancreatic neuro-endocrine tumours, 60% of stomach carcinoids, 55% of ileal carcinoids, 23% of appendiceal carcinoids and 86% of rectal carcinoids respectively in more than 10% tumour cells. No pituitary tumours displayed pS2 immunoreactivity. pS2 transcript was also detected in lung carcinoid and carotid body tumours by Northern-blot analysis. There was a statistically higher incidence of pS2 expression in carcinoid tumours of the ileum and rectum than in those of the appendix. No association was observed between pS2 expression and the occurrence of the carcinoid syndrome; nor was any correlation observed between the occurrence of pS2 immunoreactivity and that of ER. Our results suggest that the expression of the pS2 protein in a wide spectrum of neuro-endocrine tumours may be implicated in the pathogenesis and progression of some neuro-endocrine tumours in an oestrogen-independent pathway. PMID- 9221804 TI - E-cadherin expression in human epithelial ovarian cancer and normal ovary. AB - The ovarian surface epithelium (OSE) is the origin of the majority of human ovarian cancers. These adenocarcinomas are characterized by initial local growth followed by spreading into the peritoneal cavity at later stages of tumor progression. The cell-adhesion molecule E-cadherin (E-cad) plays an important role in maintaining tissue integrity. Disappearance or impaired function of E-cad have often been associated with tumor formation and invasion in vivo and in vitro. The cell-specific expression of E-cad was investigated in normal human ovaries (n = 12), in benign (n = 5) and borderline (n = 4) ovarian epithelial tumors and in adenocarcinomas of different stages and histological grades (n = 18), by immunohistochemistry and immunoblotting. An ovarian cancer cell line (NIH OVCAR3) was used as a reference. The epithelial origin of the cells was confirmed with cytokeratin (AE1/AE3) staining. In normal ovaries, the expression of E-cad was limited to inclusion cysts or deep clefts lined with OSE, whereas no staining of the OSE could be demonstrated at the surface of the ovary. In contrast, benign and borderline tumors uniformly expressed E-cad. This was observed in malignant tumors of all stages despite their degree of differentiation. E-cad was also present in metastasis from such tumors. The cell-specific expression of E-cad in inclusion cysts of normal ovaries and in epithelial layers of borderline tumors indicates a role for E-cad in the early events of the progression to a malignant phenotype. E-cad was not downregulated in later stages of ovarian cancer progression. PMID- 9221805 TI - Disease expression in Swiss hereditary non-polyposis colorectal cancer (HNPCC) kindreds. AB - The genetics of Hereditary Non-Polyposis Colorectal Cancer (HNPCC) has recently been established and found to be associated with DNA mismatch repair deficiency. As the molecular basis of this syndrome does not appear to predict any particular disease, we compared families selected according to the "Amsterdam" criteria (AC) against families that were selected because of an aggregation of colonic and extracolonic malignancies (EC), all of which have been observed in HNPCC families. A comparison of the 2 groups revealed that there were significant differences between them. Age at disease onset for both groups was 20-30 years younger than in the general population; however, a normal age distribution was observed for the AC group whereas for the EC group a bimodal distribution was apparent. The prognosis for both groups together did not differ from that of the general population; however, if split, the AC group had a significantly better outcome than the EC group. Furthermore, dividing the AC group into hMSH2- and hMLH1-linked families revealed that there was no difference in severity of disease between these 2 groups with respect to survival and mean age of disease onset. Both the AC and EC groups displayed a similar tumour spectrum with a virtually identical tumour distribution. A significant finding was the over representation, of brain tumours in this family set, which comprised the third most common malignancy after endometrial and stomach cancer. PMID- 9221806 TI - Detection of gastric mucins (M1 antigens) in cyst fluid for the diagnosis of cystic lesions of the pancreas. AB - Mucinous cystic tumors of the pancreas must be distinguished from other cystic lesions because of their potential malignancy. Our purpose was to assess the reliability of gastric M1 mucin analysis in the fluid of cystic lesions of the pancreas in comparison or association with carcinoembryonic antigen. M1 mucin and carcinoembryonic antigen were measured in cyst fluid obtained preoperatively by fine-needle aspiration. The lesions consisted of 12 serous cystadenomas, 9 mucinous cystadenomas, 8 cystadenocarcinomas and 6 intraductal mucinous hypersecreting neoplasms. Thirty pancreatic pseudocysts complicating well documented chronic pancreatitis were also examined. In addition, M1 mucins were localized by immunoperoxidase staining in fetal and normal adult pancreas and in mucinous and serous tumors. Carcinoembryonic values of > 20 ng/ml and M1 mucin values of > 50 U M1/ml represented 82 and 78% sensitivity, respectively, as well as 100% specificity for distinguishing mucinous lesions from serous cystadenomas; the sensitivity for this purpose was 100% using these criteria in combination. Carcinoembryonic antigen values of > 300 ng/ml and M1 mucin values of > 1,200 U M1/ml represented 56 and 30% sensitivity, respectively, as well as 100% specificity for distinguishing mucinous lesions from pseudocysts; the sensitivity for this purpose was 60% using these criteria in combination. By immunohistology, M1 mucins were detected in the wall of mucinous lesions but not in fetal and normal adult pancreas and in serous cystadenomas. Measurement of M1 mucin antigen in cyst fluid could thus improve the diagnosis of mucinous cystic lesions of the pancreas. PMID- 9221807 TI - Deletion analysis at the DEL-27, APC and MTS1 loci in bladder cancer: LOH at the DEL-27 locus on 5p13-12 is a prognostic marker of tumor progression. AB - Inactivation of relevant tumor-suppressor genes by allelic or homozygous deletion is a characteristic event in tumor cells. Here, the prognostic value of allelic deletions on 5p13-12 at the putative del-27 tumor-suppressor locus and in the APC tumor-suppressor gene on 5q21, as well as homozygous deletions of the MTS1 (p16INK4, CDKN 2) tumor-suppressor gene on 9p21 was assessed in 87 bladder cancers using microdissection and PCR-based assays. Tumor-specific LOH was detected in 10 of 38 (26%, del-27), and 15 of 30 (50%, APC) informative specimens. Homozygous deletion of the MTS1 gene was detected in 33% of 84 tumors investigated. These deletion frequencies implicate the 3 tumor-suppressor regions in the genesis of transitional-cell carcinoma. In contrast to deletions of the APC or MTS1 genes, LOH at the del-27 locus correlated with tumor progression. This suggests that loss of the putative tumor-suppressor gene DEL-27 is involved in an aggressive behavior of the tumor cells and appears to be a prognostic marker for the clinical outcome of patients with transitional-cell carcinoma. PMID- 9221808 TI - Endothelial and epithelial expression of sialyl Lewis(x) and sialyl Lewis(a) in lesions of breast carcinoma. AB - Tumor cells can invade and generate metastasis via either lymphatics or blood vessels. When tumor cells are circulating in the blood, they must adhere to the vessel wall, which is lined by endothelium, before they can extravasate and form new metastases. Several families of adhesion molecules have been identified to play a role in the extravasation cascade. Selectins and their sialyl Lewis(x) and/or sialyl Lewis(a) (sLe(x) and sLe(a), respectively) containing ligands play an initiating role in this cascade; we have now analyzed their role in the generation of metastatic breast carcinoma lesions. We examined expression of endothelial E- and P-selectin, expression of epithelial and endothelial sLe(x) and sLe(a) normal tissues compared with primary and metastatic breast in carcinoma lesions within individual patients. While normal breast epithelial cells do not express sLe(x) or sLe(a), epithelial expression of these oligosaccharide epitopes was enhanced in primary breast carcinoma lesions. Furthermore, epithelial expression levels of sLe(x) and/or sLe(a) were even higher in most patients (9 of 12) who had metastatic compared with primary lesions. We show that endothelia in primary lesions express more sLe(x) than in normal tissue and that metastatic lesions express even higher amounts of sLe(x) compared with primary lesions. The expression of P- and E-selectin was also greatly enhanced in tumor-bearing tissue compared with normal tissue. Our data support the hypothesis that while they are circulating in the blood, sLe(x)- and/or sLe(a)-expressing carcinoma cells have a higher probability for extravasation at sites where the endothelium expresses E- and P-selectin and for generation of new metastases. PMID- 9221809 TI - Expression of the c-Met/HGF receptor in human breast carcinoma: correlation with tumor progression. AB - Hepatocyte growth factor/scatter factor (HGF/SF) induces cell motility and tissue remodeling of various epithelial cells through its receptor, the product of the proto-oncogene c-met. High levels of HGF/SF have been correlated with poor prognosis in human breast carcinoma. In this study, we examined the expression of the c-Met receptor in human breast-carcinoma cells in vivo and in cultured cell lines. Immunohistochemical analysis of biopsy samples of human breast carcinoma indicated that, in normal mammary gland, c-Met is localized in the ductal epithelium. The level of expression of c-Met in primary carcinomas was maintained in autologous metastatic lymph-node lesions in some cases, and in other cases was elevated. Frequently there was evidence of heterogeneity in cellular expression of c-Met within individual tumors, suggesting that micro-environmental factors may regulate receptor expression. In an analysis of a panel of human breast carcinoma cell lines, we found that moderately differentiated cell lines did not express detectable levels of c-Met and were not responsive to HGF. In contrast, poorly differentiated and invasive cell lines did express high levels of the receptor and responded to HGF by increased motility and invasiveness. Sensitivity to HGF/SF also correlated with expression of the c-Met 9-kb mRNA. No correlation was found between gene copy number and the expression level of c-Met protein or mRNA. When the moderately differentiated and c-Met-negative T47D cell line was transfected with c-DNA for c-met, the transfectants showed delayed cell scattering and migratory response to HGF. Thus, over-expression of c-Met in moderately differentiated carcinoma cells may be one of several attributes that contribute to an invasive phenotype during the progression of breast cancer. PMID- 9221811 TI - Analysis of MT1-MMP and MMP2 expression in human gastric cancers. AB - Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a presumed activator of MMP2, which is one of the major proteinases in tumor cell invasion. In this study, we determined the clinico-pathologic significance of MT1-MMP expression in 68 human gastric carcinomas. The tumor-normal ratio (T/N ratio) of MTI-MMP expression was determined by reverse transcription-polymerase chain reaction analysis. To visualize the localization of MT1-MMP, an immunohistochemical study was performed. In addition, a gelatin zymography was done to examine the activation ratio of MMP2, and a correlation between MT1-MMP expression and activation of MMP2 was studied. The expression of MT1-MMP mRNA was higher in tumor tissue than in corresponding normal tissue in most cases. The mean value of the T/N ratio was 4.8. Twenty cases with T/N > or = 4.8 showed significantly deeper invasion and higher frequency of lymph node metastasis than 48 cases with T/N < 4.8. MT1-MMP expression was an independent factor influencing both tumor invasion of the gastric wall and lymph node metastasis. Although MT1-MMP expression was not an independent prognostic factor, the patients with T/N > or = 4.8 showed a significantly worse prognosis than those with T/N < 4.8. An immunohistochemical study demonstrated that MT1-MMP expression was mainly recognized in the tumor cells. There was a significant correlation between MT1 MMP expression and activation of MMP2. Our findings suggest that: 1) the expression of MT1-MMP may influence prognosis via tumor invasion of the gastric wall and lymph node metastasis, and 2) MT1-MMP activation of MMP2 may be clinically relevant in gastric carcinoma tumors. PMID- 9221810 TI - Overexpression of cyclin D1 and p53 associated with disease recurrence in colorectal adenocarcinoma. AB - Multiple genetic changes occur during the evolution of normal cells into cancer cells. It has been reported that both cyclin D1 and p53 genes play major roles in oncogenesis and/or cell cycle control in various cancers. In this study, we examined the overexpression of cyclin D1 and p53 by the immunohistochemical method and investigated the correlation between expression of these antigen and prognosis in patients with colorectal adenocarcinoma. Disease-free survival was significantly lower in the patients with cyclin D1-strongly positive tumors than in those with cyclin D1-negative tumors. Similarly, disease-free survival of the patients with p53-strongly positive tumors was significantly lower than that of those with p53-negative tumors. Moreover, multivariate analysis indicated that both cyclin D1 and p53 overexpression are independent prognostic factors in patients with colorectal adenocarcinoma. In conclusion, both cyclin D1 and p53 overexpression may be useful predictors of disease recurrence in patients with colorectal adenocarcinoma. PMID- 9221812 TI - Genomic deletions in the BRCA1, BRCA2 and TP53 regions associate with low expression of the estrogen receptor in sporadic breast carcinoma. AB - Sporadic breast carcinoma is associated with multiple genetic alterations. The clinical relevance of these alterations, however, needs further clarification. In the present study we analyzed 266 spontaneously arising breast carcinomas for allelic losses in the BRCA1 and TP53 regions on chromosome 17, the BRCA2 region on chromosome 13, the ATM (mutated in ataxia-telangiectasia) region on chromosome 11 and on the chromosomal arms 7q and 16q. In addition the following clinical and pathological parameters were evaluated: age at diagnosis, tumor size, presence or absence of regional and distant metastases, hormone-receptor status, histopathological classification and tumor grading. The analysis of genetic and clinical observations revealed significant associations: absence of expression of the estrogen receptor was linked to a high rate of allelic losses of markers in the BRCA1, TP53 and BRCA2 regions. Expression of the progesterone receptor coincided with allelic loss on the long arm of chromosome 16. High-grade malignant lesions and ductal differentiation were frequently associated with allelic losses in the proximal portion of chromosome 17q. The accumulation of multiple allelic deletions was linked to high-grade malignant tumors, to tumor size, and to loss of expression of the estrogen receptor. Our data point to a relationship between clinically relevant prognostic factors and specific genomic deletions in the BRCA1, BRCA2 and TP53 region. PMID- 9221813 TI - Vascularity and prognosis of metastatic melanoma. AB - The clinical role of vascularity was examined in metastatic melanoma, analyzing the correlation of the blood vessel density and prognosis. Our study included 51 specimens of metastatic melanoma tissue samples from 31 patients treated with combined chemo-immunotherapy. PECAM-1 (CD31) was used for assessing vascularity by immunohistochemical staining. On the basis of blood vessel counts, patients were classified into 2 main groups: low and high vascularity. A higher blood vessel density was found to be associated with shorter survival, estimated from the primary diagnosis of the disease (38 months), compared with patients with low blood vessel counts (68 months). A similar tendency was observed when vascularity was correlated to the survival period after the detection of the first metastases (13 vs. 30 months) and with survival since the initiation of chemo-immunotherapy (8 vs. 16 months). When vascularity and some common prognostic factors, such as age, sex, DNA ploidy and WHO tumor response, were used for a Cox multivariate analysis, vascularity turned out to be the most significant independent prognostic factor. Our results suggest that counting the blood vessels identified by immunohistochemical staining for the endothelial cell-specific CD31 is a powerful predictor for prognosis in patients with metastatic melanoma and should be considered when selecting patients for therapy. PMID- 9221814 TI - Reduced survival in patients with stage-I non-small-cell lung cancer associated with DNA-replication errors. AB - To better understand whether replication-error-type instability (RER+) is a frequent genetic alteration event in surgical-pathologic stage-I non-small-cell lung cancer (NSCLC) and identify whether it constitutes an independent prognostic parameter, we examined 35 surgical-pathologic stage-I-NSCLC patients with complete follow-up in all cases for at least 49 months. The tumor samples and the paired histopathologically normal lung samples for each patient were analyzed for 8 microsatellite markers located at chromosomes 3p and 2p to investigate microsatellite alterations such as RER+ and loss of heterozygosity (LOH). Single strand-conformation-polymorphism analysis for detection of p53 and k-ras gene mutations was also carried out. Genetic data were correlated with clinical outcome and histopathologically established prognostic factors. RER+ at one or both chromosomes was identified in 24 of the 35 patients; 9 patients showed LOH. A statistically significant correlation was found between RER+ and poor prognosis (p = 0.001). Furthermore, RER+ proved to be an independent factor that predicted decreased survival, ranking first, followed by visceral pleural invasion. A trend towards worse survival was strongest in the group of patients with tumor size greater than 3 cm (T2). Patients with other genetic abnormalities, such as K-ras mutations, p53 mutations or LOH, had prognoses similar to those of patients without such aberrations. The data suggest that RER+ is common in NSCLC, that it may provide important prognostic information in stage-I NSCLC and serve as a useful marker for relapse-risk assessment in operable NSCLC patients. PMID- 9221815 TI - Heparan sulfate proteoglycan expression in human lung-cancer cells. AB - Heparan sulfate (HS) functions as a co-factor in several signal-transduction systems that affect cellular growth, differentiation, adhesion and motility. HS, therefore, may also play a role in the malignant transformation of cells, tumor growth, cell invasiveness and the formation of tumor metastases. To explore this hypothesis, we analyzed the expression of HS and heparan sulfate proteoglycan (HSPG) in histological sections of human lung-cancer tissues and assayed for the presence of HSPGs in extracts of human lung-cancer cell lines, using a panel of native HS-, delta-HS- and HSPG (syndecan, glypican, CD44 and perlecan) core protein-specific monoclonal antibodies. Compared to normal epithelia, non-small cell lung carcinomas, particularly poorly differentiated tumors, often expressed reduced amounts of the major cell surface-associated HSPGs (most consistently of syndecan-1). CD44 or CD44-variant proteins, in contrast, were found on all tumor cells, irrespective of their differentiation. Perlecan, a matrix-associated HSPG found in the basement membrane of normal bronchial epithelium, was consistently undetectable in invasive bronchogenic carcinomas. Staining reactions for native HS were consistently reduced in squamous-cell lung carcinomas, in the cells in contact with the stroma and in the less differentiated areas of these tumors. Reactions for delta-HS, however, were not reduced, suggesting a structural change in the HS of these tumor cells. Poorly differentiated adenocarcinomas, in contrast, yielded strong HS and delta-HS reactions. Marked differences in HSPG expression also were observed among various non-small-cell lung carcinoma cell lines. Our results suggest that poorly differentiated lung tumors have markedly altered patterns of HSPG expression, which may contribute to their invasive phenotype. PMID- 9221816 TI - Prognostic significance of Bcl-2 expression and p53 nuclear accumulation in colorectal adenocarcinoma. AB - The products of bcl-2 and p53 genes are involved in the regulation of apoptosis and proliferation and have been associated with prognosis in several malignancies, including colorectal adenocarcinoma. Although 2 European studies have reported a prognostic significance of Bcl-2 expression in colorectal adenocarcinomas, a study from the United States did not observe such an association. Therefore, we used immunohistochemistry to evaluate the prognostic significance of Bcl-2 expression, p53 nuclear accumulation and their concomitant expression in 134 US patients with colorectal adenocarcinoma. Antigen retrieval was required for adequate detection of Bcl-2 expression. Fifty percent of the colorectal tumors were classified as expressing Bcl-2, and Bcl-2 expression was associated with longer patient survival. Antigen retrieval was not necessary for detecting nuclear accumulation of p53 by immunohistochemistry. Nuclear accumulation of p53 was detected in 44% of colorectal adenocarcinomas and was associated with decreased patient survival. Tumors that did not express detectable levels of Bcl-2 but exhibited nuclear accumulation of p53 were associated with the shortest patient survival (log rank, p = 0.001). Multivariate Cox regression analysis demonstrated that Bcl-2 expression (p = 0.018), p53 nuclear accumulation (p = 0.024) and regional lymph-node metastasis (p = 0.005) were independent prognostic factors. Although a trend toward an inverse correlation between Bcl-2 and p53 expression was observed, the prognostic value of Bcl-2 expression was independent of p53 status. Thus, assessment of both Bcl-2 and p53 status may be valuable in predicting the prognosis of patients with colorectal adenocarcinomas. PMID- 9221817 TI - Is there a lung-cancer susceptibility gene? PMID- 9221819 TI - Inhalation of high concentrations of low toxicity dusts in rats results in impaired pulmonary clearance mechanisms and persistent inflammation. AB - This study was carried out to assess the time course of pulmonary clearance impairment and persistence of inflammation following high-dose inhalation exposures to titanium dioxide (TiO2) or carbonyl iron (CI) particles. Male rats were exposed to air, TiO2 or CI particles 6 hr/day, 5 days/week, for 4 weeks at concentrations of 5, 50, and 250 mg/m3 and evaluated at selected intervals through 6 months postexposure. Indices of pulmonary inflammation as well as alveolar macrophage clearance functions (i.e., morphology, in vivo and in vitro phagocytosis, and chemotaxis), cell proliferation, and histopathology endpoints were measured at several postexposure time periods through 6 months. In addition, amounts of TiO2 or CI in lungs and tracheobronchial lymph nodes were measured to allow an evaluation of particle clearance and translocation patterns. Four-week exposures to TiO2 or CI particles at concentrations of 250 mg/m3 resulted in lung burdens of 12 mg titanium and 17 mg iron, respectively, with particle retention half-times ranging from 68 days for 5 mg/m3 TiO2 to approximately 330 days for 250 mg/m3. The impact of this TiO2 dust load and similar lung burdens of CI particles produced a sustained pulmonary inflammatory response measured through a period of 3-6 months postexposure concomitant with increases in BrdU cell labeling of terminal airway and pulmonary parenchymal cells. The impairment of particle clearance mechanisms was accounted for by deficits in in vitro phagocytic and chemotactic potential of alveolar macrophages recovered from the lungs of high-dose, TiO2- or CI-exposed rats. Free granular pigment (TiO2 or CI) was present on the hypertrophic mucosal surfaces of bronchioles and bronchi, and particle-laden macrophages, found individually, were numerous throughout alveoli and within lymphoid tissues immediately after exposure. Aggregates of particle laden macrophages were present within alveoli and alveolar ducts from 1 week postexposure through the entire 6-month recovery period. Macrophage accumulations increased in size and number from 1 week through 1 month postexposure and then appeared to remain constant through the remaining 5-month postexposure period. Minimal cellular hypertrophy and hyperplasia were evident at alveolar duct bifurcations adjacent to macrophage aggregates, and this effect was most prominent at 3 to 6 months postexposure. The results of this study clearly demonstrate that exposure to high dust concentrations of two different innocuous particle types produced sustained pulmonary inflammation, enhanced proliferation of pulmonary cells, impairment of particle clearance, deficits in macrophage function, and the appearance of macrophage aggregates at sites of particle deposition. In addition, the mass deposition rate determination appears to be a less sensitive indicator of "overload" when compared to biomarkers of pulmonary toxicity, such as macrophage function and cellular inflammation and proliferation indices. PMID- 9221818 TI - Species and strain differences in the hepatic cytochrome P450-mediated biotransformation of 1,4-dichlorobenzene. AB - Our goal was to characterize possible species and strain differences in the hepatic microsomal biotransformation of 1,4-dichlorobenzene (1,4-DCB). Experiments compared extent of labeled 1,4-DCB conversion to oxidized metabolites, glutathione conjugates, and covalently bound metabolites by hepatic microsomes from humans, from male B6C3F1 mice, and from males of three rat strains (Fischer 344, Sprague-Dawley (SD), and Wistar). These rodents were selected for comparison because of their dissimilar responses to 1,4-DCB, notably, hepatocarcinogenicity in the B6C3F1 mouse but not the Wistar or Fischer rat, and nephrotoxicity and carcinogenicity in the Fischer rat. The species rank order for total in vitro conversion of 1,4-DCB was mouse > rat >> human. Conversion by microsomes from Fischer and Wistar rats was similar, whereas SD rats showed less biotransformation than the other two strains. Microsomes from the mouse produced most of the reactive metabolites as indicated by covalent binding to macromolecules (>20% of total metabolites formed). This covalent binding by mouse microsomes was extensively inhibited by ascorbic acid (AA), with a concomitant increase in hydroquinone formation, indicating an important role for benzoquinones as reactive metabolites. Phenobarbital pretreatment of rats enhanced the in vitro conversion of 1,4-DCB and the amount of covalent binding. Covalent binding for all rat microsomes was partly (33-79%) inhibited by AA. Addition of glutathione (GSH) plus AA further diminished the covalent binding with concomitant increased formation of the GSH-conjugated epoxide. Human microsomes produced the least reactive metabolites, with the majority (>70%) of this covalent binding prevented by GSH addition. The observed species differences, notably the more pronounced biotransformation of 1,4-DCB to reactive species including benzoquinones, could be factors in this compound's liver carcinogenicity in B6C3F1 mice but not other rodent species. PMID- 9221820 TI - Studies on the mechanism of uroporphyrinogen decarboxylase inhibition in hexachlorobenzene-induced porphyria in the female rat. AB - Hexachlorobenzene (HCB)-induced porphyria occurs in female, but not male, rats after a delay of 35 days following HCB treatment. Uroporphyrinogen decarboxylase (UROD) inhibition has been proposed as a primary causative event. To determine whether there also exists a delay phase and a sexual dimorphism for UROD inhibition, groups of male and female rats were given HCB (100 mg/kg/day) from Days 1 to 5. Hepatic uroporphyrin III was markedly increased only after Day 33. Liver cytosol UROD activity in HCB-treated female rats with porphyria at Days 33, 40, 47, 54, and 100 was decreased by over 70% compared to concurrent control, whereas treated male rats as well as nonporphyric female rats had UROD activity comparable to control levels at Days 6, 12, 19, 26, 33, 40, 47, and 54. Level of immunoreactive UROD in cytosol of porphyric rats was not modified by HCB. No gender-related differences in liver cytosol radiolabel level ([14C]HCB given as the fifth dose) were found at Days 6 and 30. Chromatography of liver cytosol showed nonspecific binding of radiolabel to proteins for males, porphyric and nonporphyric females, and loss of UROD activity did not correlate with the amount of radiolabel in the UROD-containing fractions. Thus, the gender-specific decrease in UROD activity observed when porphyria develops in female rats (delay of about 4 weeks), as well as the persistence of low activity and porphyria for months, suggests that UROD inhibition was causally related to porphyria. PMID- 9221821 TI - Comparative in vitro skin absorption and metabolism of coumarin (1,2-benzopyrone) in human, rat, and mouse. AB - The in vitro percutaneous absorption and skin metabolism of coumarin (1,2 benzopyrone) was studied in metabolically viable human, rat (F344), and mouse (CD1 and DBA/2) skin. Following application of [14C]coumarin (3.7 microg/cm2; 0.02% in ethanol) to unoccluded skin in flow-through diffusion cells of a skin absorption model (SAM), the absorption through the skin into the receptor fluid at 72 hr was rapid and extensive in all species, reaching (mean +/- SD) 50.4 + 9.1% of the applied dose in human, 51.3 +/- 7.3% in rat, and 44.9 +/- 13.5% in mouse. When the skin was occluded immediately after exposure, the extent of absorption at 72 hr was enhanced in all species. At 72 hr, substantial amounts of [14C]coumarin were found in unoccluded mouse skin (31.7 +/- 13.6%), with less in human (10.2 +/- 6.5%) and rat (12.7 +/- 5.0%) tissue. When occluded, the skin residues at 72 hr were 10.4 +/- 11.7% (mouse), 8.5 +/- 3.9% (human), and 11.9 +/- 7.5% (rat). The absorption of coumarin through rat skin into the receptor fluid over 72 hr was linearly related to the applied dose (r2 = 0.998 unoccluded skin; r2 = 0.999 occluded skin) over the dose range 3.7 to 378.7 microg/cm2. The nature and extent of cutaneous metabolism was studied following (i) topical application for 24 hr to human, rat, and mouse skin in the SAM system; (ii) incubation at 37 degrees C for up to 6 hr with human, rat, and mouse whole skin homogenates; and (iii) incubation at 37 degrees C for up to 24 hr with freshly isolated and cultured human epidermal keratinocytes. HPLC and GCMS analyses of skin extracts and receptor fluid confirmed that, in all three species, only the parent compound, coumarin, was present at all times from 10 min to 24 hr. These data indicate that topically applied coumarin is rapidly and extensively absorbed through human, rat, and mouse skin, and that the compound remains metabolically unchanged during absorption. These observations may have implications for the safe and effective use of coumarin in consumer products which come into contact with the skin and as a topical therapeutic agent. PMID- 9221822 TI - The stoichiometry of protection against soman and VX toxicity in monkeys pretreated with human butyrylcholinesterase. AB - Bioscavengers of organophophates (OP) have been examined as potential substitutes for the currently approved drug treatment against OP toxicity. The present work was designed to assess the ability of butyrylcholinesterase, purified from human serum (HuBChE), to prevent the toxicity induced by soman and VX in rhesus monkeys. The consistency of the data across species was then evaluated as the basis for the extrapolation of the data to humans. The average mean residence time of the enzyme in the circulation of monkeys following an intravenous loading was 34 hr. High bioavailability of HuBChE in blood (>80%) was demonstrated after intramuscular injection. A molar ratio of HuBChE:OP approximately 1.2 protected against an i.v. bolus injection of 2.1 x LD50 VX, while a ratio of 0.62 was sufficient to protect monkeys against an i.v. dose of 3.3 x LD50 of soman, with no additional postexposure therapy. A remarkable protection was also seen against soman-induced behavioral deficits detected in the performance of a spatial discrimination task. The consistency of the results across several species offers a reliable prediction of both the stoichiometry of the scavenging and the extent of prophylaxis with HuBChE against nerve agent toxicity in humans. PMID- 9221823 TI - Cytochrome P450 metabolism of estradiol in hamster liver and kidney. AB - Estradiol induces kidney tumors in Syrian hamsters. The elevated conversion of estradiol to 4-hydroxylated metabolites in kidney compared to the predominant 2 hydroxylation in liver and other organs, where tumors are not induced by this treatment, has been proposed to be the basis of estrogen-induced carcinogenesis. In this study, we examined the hepatic and renal enzymes catalyzing the formation of catecholestrogens to understand the differences in estrogen metabolism in these organs. In liver, 2-hydroxylation of estradiol is the major metabolic pathway with 4-hydroxylation a minor by-product and with the formation of both catechols responding coordinately to the same inhibitors. Western blot analysis and inhibition studies suggest that the major form catalyzing hepatic estrogen 2 hydroxylation is a member of the CYP3A family, as previously observed with rat liver microsomes, and that 4-hydroxylation is a by-product of this metabolism. In the kidney, 4-hydroxylation of estradiol appears to be catalyzed by more than one enzyme according to the Eadie-Hofstee analysis. Both 2- and 4-hydroxylation in the kidney are affected differentially by inhibitors and are induced by beta napthoflavone. Western blots of renal microsomes reveal that CYP1A2 is induced whereas CYP1A1 is detectable in kidney, but not induced by this treatment. Finally, a part of the 2-hydroxylation and a small part of the 4-hydroxylation by kidney microsomes may be catalyzed by a member of the CYP3A family, since these reactions are partially inhibited by CYP3A inhibitors such as progesterone and other progestins, although renal enzyme levels are much lower than those in the liver as revealed by Western blot. Our data suggest that estrogen 2-hydroxylation in the hamster kidney is catalyzed by members of the CYP1A and CYP3A families, which also contribute to 4-hydroxylation. The majority of 4-hydroxyestradiol formation in the hamster kidney may be catalyzed by a form(s) of the newly discovered CYP1B family that has yet to be characterized. PMID- 9221824 TI - Inhalation of toluene diisocyanate is associated with increased production of nitric oxide by rat bronchoalveolar lavage cells. AB - Isocyanates are used commercially, particularly in the manufacture of polyurethane coatings and foam. These compounds can pose an occupational health hazard since there is a risk of respiratory disease following isocyanate exposure. The purpose of the present study was to investigate whether a single, sublethal isocyanate inhalation is associated with increased production of the free radical nitric oxide (NO). Mature male Sprague-Dawley rats were exposed to air or toluene diisocyanate (TDI; 2 ppm) for 4 hr. Indices of pulmonary function were assessed before and after exposure to TDI fumes. At 20 hr postexposure, bronchoalveolar lavage cells (BALC) and fluid were harvested. NO synthase (NOS) dependent reactive species production by alveolar macrophages was assessed by determining N(omega)-nitro-L-arginine methyl ester-inhibitable chemiluminescence following stimulation with unopsonized zymosan. Northern blot analysis was used to index inducible NOS mRNA levels in BALC, while nitrite and nitrate (NOx) levels were measured to determine NOx levels in the lavage fluid and the production of NO by cultured adherent BALC was indexed by measuring nitrite levels. Exposure to aerosolized TDI was associated with an increase in the number of alveolar macrophages, lymphocytes, and polymorphonuclear leukocytes harvested by bronchoalveolar lavage, relative to that from air-exposed rats. NOx levels in the lavage fluid and NOS-dependent production of reactive species by alveolar macrophages were increased following TDI exposure. In addition, inducible NO production by BALC (i.e., mRNA levels and nitrite levels in BALC conditioned media) was elevated following TDI treatment. These findings indicate that pulmonary inflammatory responses induced by TDI exposure are associated with increases in inducible NO production. Therefore, the potential role of NO in the initial pulmonary response to TDI exposure warrants further investigation. PMID- 9221825 TI - Effects of cadmium on bone: an in vivo model for the early response. AB - Cadmium (Cd) exposure induces bone resorption in vitro and in vivo that can lead to low bone mass and increased incidence of fracture. We have developed an animal model for following the early skeletal response to Cd. A low-calcium (but not calcium-deficient) diet is used to increase gastrointestinal absorption of calcium so that the endogenous fecal calcium excretion is essentially the total fecal calcium excretion. The bone response is followed by quantitation of stable fecal calcium and does not require a radioactive label. After mice were adjusted to a low-calcium diet, Cd was administered by a single gavage and fecal calcium was monitored to determine the magnitude of the calcium release from bone. Fecal calcium excretion (microg Ca/hr; mean +/- SE) remained at the background level for 8 hr (13.6 +/- 1.8, n = 18) but increased during the 8- to 24-hr and 24- to 56-hr collection periods (43.8 +/- 6.8, n = 12; 50.75 +/- 3.7, n = 6, respectively). The bone response was transient and dropped to nearly background levels during the 56- to 104-hr collection period. Blood calcium levels were normal throughout the time course. Bone resorption occurred at Cd levels of 7.9 +/- 0.7 microg/liter blood (mean +/- SE, n = 6), which is in the range of occupational exposure levels. The transient nature of the bone response contrasted to the slow but continuing rise observed in blood Cd levels. These results suggest that a threshold level of Cd is required for a bone response but that chronic levels of Cd in blood do not necessarily indicate the occurrence of continuous active bone resorption. This model can be used to probe early gene changes (prior to the bone response) that may be occurring in response to Cd exposure. PMID- 9221827 TI - Pharmacokinetics of ochratoxin A and its metabolites in rats. AB - Ochratoxin A (OA) is a mycotoxin that is produced on moist grain. It is commonly found in the blood of swine in western Canada and is a potent nephrotoxic, carcinogen, and immunosuppressive agent. The pharmacokinetic characteristics of six analogs of OA including OA, OB (OA without chloride), OC (OA ethyl ester), and some metabolites, such as O alpha (OA without phenylalanine), OA-OH (hydroxylated OA), and a newly discovered form of OA, OP-OA (lactone opened ring of OA), were investigated in rats after a single intravenous administration of the compounds. All of the ochratoxin analogs were distributed following a two compartment open model. The elimination half-lives of OA, OP-OA, O alpha, OA-OH, OB, and OC were 103+/-16, 50.5+/-2.8, 9.6+/-2.3, 6+/-0.9, 4.2+/-1.2, and 0.6+/ 0.2 hr, respectively. Total body clearance of OA, OP-OA, O alpha, OA-OH, and OB via the bile, urine, and metabolic routes were 3.1, 3.6, 40, 65, and 43 ml/hr kg, respectively. OA, OB, and O alpha were mainly cleared in the urine (> or = 48%), OA-OH in the bile (41%), and OP-OA as metabolites (43%). Metabolism accounted for 43, 44, 33, and 29% of the total clearance of OA, O alpha, OA-OH, and OB, respectively. It is concluded that OA has a long half-life and is very slowly cleared from the body and that its metabolites are cleared at a much faster rate with much shorter half-lives. Procedures should be devised to enhance the conversion in the body of OA to O alpha, OA-OH, or other metabolites as this would shorten its half-life and therefore its toxicity. PMID- 9221826 TI - Role of mitochondria and calcium ions in tributyltin-induced gene regulatory pathways. AB - Tributyltin (TBT) salts are potent skin irritants both in humans and rodents. Data in the literature indicate mitochondria as target of TBT effects. Here, we investigate the early intracellular molecular events that follow TBT treatment and the relevance of calcium ions and mitochondria in gene-regulatory signaling pathways. Confluent HEL30 cells were treated with increasing doses of TBT (0-5 microM). At different times thereafter, the level of intracellular Ca2+, the cellular oxidative activity, nuclear factor-kappaB (NF-kappaB) activation, and IL 1alpha production were measured. TBT induced a dose-related increase of intracellular Ca2+ that reached the plateau 4 min following treatment. The increase of intracellular Ca2+ was followed by an increase in cellular oxidative activity as measured by DCFH oxidation (15 min) that preceded NF-kappaB activation (30 min) and IL-1alpha production (4 hr). All these events can be almost completely abrogated by BAPTA, an intracellular Ca2+ chelator. Furthermore, the modulation of cellular oxidative activity induced by TBT observed with rotenone, an inhibitor of the electron entry from complex I to ubiquinone, or after prolonged treatment with ethidium bromide, an inhibitor of mitochondrial DNA and RNA synthesis, indicates mitochondria as an important intracellular source of reactive oxygen species. These findings indicate the rise in intracellular Ca2+ as the starting event and indicate the role of mitochondria as the source of second messenger molecules essential for TBT-induced NF-kappaB activation and IL-1alpha production. PMID- 9221828 TI - Inhibition of cholesterol synthesis by squalene synthase inhibitors does not induce myotoxicity in vitro. AB - The cholesterol-lowering HMG CoA reductase inhibitors (HMGRI), pravastatin and lovastatin, have been associated with skeletal myopathy in humans and in rats. In a previous in vitro study, HMGRI-induced changes in neonatal rat skeletal muscle cells were characterized by reversible inhibition of protein synthesis and loss of differentiated myotubes at concentrations markedly lower than those inducing enzyme leakage. Myotoxicity was determined to be directly related to inhibition of HMG CoA reductase, since mevalonate, the immediate product of HMG CoA reductase metabolism, abrogated the drug-induced changes. Farnesol, geranylgeraniol, and squalene are metabolites of mevalonate. Squalene, formed from farnesol by squalene synthase, is the first metabolite solely committed to cholesterol synthesis. In contrast, geranylgeraniol, formed by the addition of an isoprene group to farnesol, is the first metabolite uncommitted to cholesterol synthesis. The objective of the present study was to determine the role of inhibition of cholesterol synthesis in HMGRI-induced in vitro myotoxicity. HMGRI treated neonatal rat skeletal muscle cultures were supplemented with farnesol and geranylgeraniol, and in another study, muscle cultures were exposed to two squalene synthase inhibitors (SSI), BMS-187745 and its prodrug ester, BMS-188494. Endpoints evaluated for both studies included protein synthesis ([3H]leucine incorporation), total cellular protein (a measure of cell loss), intra- and extracellular lactate dehydrogenase activity (a measure of membrane integrity), cholesterol biosynthesis ([14C]acetate incorporation), and morphology. HMG CoA reductase inhibitor-induced morphologic changes and inhibition of protein synthesis were significantly ameliorated by supplementation with farnesol and geranylgeraniol. In contrast to HMGRI-induced in vitro myotoxicity, SSI induced an irreversible, minimal cytotoxicity at close to maximum soluble concentrations. These results indicate that depletion of metabolites of geranylgeranyl pyrophosphate, and not inhibition of cholesterol synthesis, is the primary cause of HMG CoA reductase-induced myotoxicity. PMID- 9221829 TI - HMG CoA reductase inhibitor-induced myotoxicity: pravastatin and lovastatin inhibit the geranylgeranylation of low-molecular-weight proteins in neonatal rat muscle cell culture. AB - In previous studies, inhibition of cholesterol synthesis by HMG CoA reductase inhibitors (HMGRI) was associated with myotoxicity in cultures of neonatal rat skeletal myotubes, and rhabdomyolysis in rats, rabbits, and humans in vivo. In vitro myotoxicity was directly related to HMGRI-induced depletion of mevalonate, farnesol, and geranylgeraniol, since supplementation with these intermediate metabolites abrogated the toxicity. Both farnesol and geranylgeraniol are required for the posttranslational modification, or isoprenylation, of essential regulatory proteins in mammalian cells. The objective of the present study was to measure changes in protein isoprenylation in cultured neonatal rat skeletal muscle cells exposed for 24 hr to increasing concentrations of pravastatin or lovastatin. Proteins were labeled with [3H]mevalonate, [3H]farnesyl pyrophosphate (FPP), or [3H]geranylgeranyl pyrophosphate (GGPP), and then separated by SDS-PAGE and quantitated by scintillation counting and densitometry of autoradiographs. Mevalonate and FPP labeling of the majority of proteins increased in a concentration-dependent manner, even at concentrations greater than 2 microM lovastatin and 25 microM pravastatin that completely inhibited cholesterol synthesis. In contrast, mevalonate and FPP labeling of three protein bands with molecular weights of 26.6, 27.7, and 28.9 kDa was markedly inhibited at concentrations higher than 1 microM lovastatin and 400 microM pravastatin, which inhibited protein synthesis and disrupted myotube morphology after longer exposures in a previous study. In contrast, these proteins were equally well labeled by GGPP at all HMGRI concentrations tested, suggesting that isoprenylation of the 26.9-, 27.8-, and 28.9-kDa proteins requires geranylgeraniol. The results of this study indicate that HMGRI-induced myotoxicity is most likely related to reduced posttranslational modification of specific regulatory proteins by geranylgeraniol. PMID- 9221830 TI - Hydroxylated polychlorinated biphenyls (PCBs) as estrogens and antiestrogens: structure-activity relationships. AB - The effects of structure on the estrogenicity and antiestrogenicity of hydroxylated polychlorinated biphenyls were investigated using the following estrogen-sensitive assays: competitive binding to the rat and mouse cytosolic estrogen receptor (ER); immature rat and mouse uterine wet weight, peroxidase and progesterone receptor (PR) levels; induction of luciferase activity in HeLa cells stably transfected with a Gal4:human ER chimera and a 17mer-regulated luciferase reporter gene; proliferation of MCF-7 human breast cancer cells; induction of chloramphenicol acetyl transferase (CAT) activity in MCF-7 cells transiently transfected with a full-length human ER expression plasmid and a plasmid containing an estrogen-responsive vitellogenin A2 promoter linked to a CAT reporter gene. The chemicals synthesized for this study contained a 4-hydroxy group in one ring, a 2- or 3-chloro substituent meta or ortho to the hydroxyl group, and variable substitution (2',3',4',5'-, 2',3',4',6'-, 2',3',5',6' tetrachloro and 2',4',6'-trichloro) in the chlorophenyl ring. The compounds included: 2,2',3',4',5'- (A), 2,2',3',4',6'- (B), and 2,2',3',5',6'-pentachloro- (C); 2,2',4',6'-tetrachloro-4-biphenylol (D); 2',3,3',4',5'- (E), 2',3,3',4',6'- (F), and 2',3,3',5',6'-pentachloro (G); and 2',3,4',6'-tetrachloro-4-biphenylol (H). With the exception of 2',3,4',6'-tetrachloro-4-biphenylol (H), all of the compounds competitively bound to the mouse and rat ER with relative binding affinities [compared to 17beta-estradiol (E2)] varying from 1.4 x 10(-3) to 5.3 x 10(-5). The structure-ER binding relationships for the hydroxy-PCB congeners were different in the rat and mouse, and no dose-dependent estrogenic activities were observed in the mouse or rat uterus. Several hydroxy-PCB congeners exhibited antiestrogenic activity (primarily in the mouse uterus) and two compounds, 2,2',3',5',6- and 2,2',3',4',6'-pentachloro-4-biphenylol, inhibited E2-induced uterine wet weight, PR binding, and peroxidase activity in the mouse uterus. 2,2',3',4',5'- and 2,2',3',4',6'-Pentachloro-4-biphenylol induced CAT activity in MCF-7 cells transiently transfected with the Vit-CAT plasmid; the remaining congeners did not induce CAT activity but exhibited antiestrogenic activity in MCF-7 cells cotreated with 10(-9) E2 plus 10(-5) M hydroxy-PCBs. Complementary structure-estrogenicity relationships were observed utilizing the HeLa cell luciferase induction and MCF-7 cell proliferation assays. The placement of the 2- or 3-chloro groups in the phenolic ring had minimal effects on estrogenic activity, whereas 2,4,6-trichloro- and 2,3,4,6-tetrachloro substitution in the chlorophenyl ring (B, D, F, and H) were required for this response. Substitution in the phenolic ring was also not important for structure-antiestrogenicity relationships, and the most active compounds (A, C, E, and G) contained 2',3',4',5'- and 2',3',5',6'-tetrachlorophenyl groups. Thus, structure estrogenicity/antiestrogenicity relationships for this series of hydroxy-PCBs were complex and response-specific. PMID- 9221831 TI - In utero and lactational exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin: effects on development of the male and female reproductive system of the mouse. AB - To evaluate effects of in utero and lactational 2,3,7,8-tetrachlorodibenzo-rho dioxin (TCDD) exposure on male and female reproductive system development of the mouse, the offspring of pregnant ICR mice administered 0, 15, 30, or 60 microg TCDD/kg on Gestation Day (GD) 14 were examined at the postweanling, pubertal, young adult, and adult stages of development. Dam and offspring body weights and prenatal and postnatal mortality were unaffected by TCDD exposure. The most sensitive endpoints in male offspring were decreased ventral prostate, coagulating gland, and thymus weights, accelerated eye opening, and hydronephrosis. Decreases in pituitary gland weight and epididymal sperm numbers were also found in TCDD-exposed male offspring. Testis, epididymis, and dorsolateral prostate weights, anogenital distance, latencies to testis descent and to preputial separation, and serum testosterone concentrations were unaffected. At the highest maternal TCDD dose uterus weights were decreased in female offspring evaluated during estrus and diestrus. No morphologic changes in the external genitalia of female offspring were found, nor were there alterations in ovary or pituitary gland weights. Cross-species comparisons showed that the mouse was not as sensitive to TCDD-induced developmental reproductive toxicity as the rat and hamster. Many endpoints affected by TCDD in rat and hamster offspring were either not affected or were less sensitive in mouse offspring. Endpoints of androgenic status were not affected in the mouse, decreases in accessory sex organ weights were restricted to fewer organs in the mouse, and decreases in daily sperm production were not found in the mouse. The only developmental reproductive endpoint observed in all three species was a reduction in epididymal sperm numbers. PMID- 9221832 TI - Effect of dietary cadmium on iron metabolism in growing rats. AB - Little is known regarding the interactions between iron and cadmium during postnatal development. This study examined the effect of altered levels of dietary iron and cadmium loading on the distribution of cadmium and iron in developing rats ages 15, 21, and 63 days. The uptake of iron, transferrin, and cadmium into various organs was also examined using 59Fe, [125I]transferrin, and 109Cd. Dietary cadmium loading reduced packed cell volume and plasma iron and nonheme iron levels in the liver and kidneys, evidence of the inducement of an iron deficient state. Dietary iron loading was able to reverse these effects, suggesting that they were the result of impaired intestinal absorption of iron. Cadmium loading resulted in cadmium concentrations in the liver and kidneys up to 20 microg/g in rats age 63 days, while cadmium levels in the brain reached only 0.16 microg/g, indicating that the blood-brain barrier restricts the entry of cadmium into the brain. Iron loading had little effect on cadmium levels in the organs and cadmium feeding did not lower tissue iron levels in iron loaded animals. These results suggest that cadmium inhibits iron absorption only at low to normal levels of dietary iron and that at high levels of intake iron and cadmium are largely absorbed by other, noncompetitive mechanisms. It was shown that 109Cd is removed from the plasma extremely quickly irrespective of iron status and deposits mainly in the liver. One of the most striking effects of cadmium loading on iron metabolism was increased uptake of [125I]transferrin by the heart, possibly by disrupting the process of receptor-mediated endocytosis and recycling of transferrin by heart muscle. PMID- 9221833 TI - A comparison of physiologically based pharmacokinetic model predictions and experimental data for inhaled ethanol in male and female B6C3F1 mice, F344 rats, and humans. AB - Ethanol is added to unleaded gasoline as an oxygenate to decrease carbon monoxide automobile emissions. This introduces inhalation as a new possible route of environmental exposure to humans. Knowledge of the pharmacokinetics of inhaled ethanol is critical for adequately assessing the dosimetry of this chemical in humans. The purpose of this study was to characterize the pharmacokinetics of inhaled ethanol in male and female B6C3F1 mice and F344 rats and to develop a physiologically based pharmacokinetic (PBPK) model for inhaled ethanol in mice, rats, and humans. During exposure to 600 ppm for 6 hr, steady-state blood ethanol concentrations (BEC) were reached within 30 min in rats and within 5 min in mice. Maximum BEC ranged from 71 microM in rats to 105 microM in mice. Exposure to 200 ppm ethanol for 30 min resulted in peak BEC of approximately 25 microM in mice and approximately 15 microM in rats. Peak BEC of about 10 microM were measured following exposure to 50 ppm in female rats and male and female mice, while blood ethanol was undetectable in male rats. No sex-dependent differences in peak BEC at any exposure level were observed. Species-dependent differences were found following exposure to 200 and 600 ppm. A blood flow limited PBPK model for ethanol inhalation was developed in mice, rats, and humans which accounted for a fractional absorption of ethanol. Compartments for the model included the pulmonary blood and air, brain, liver, fat, and rapidly perfused and slowly perfused tissues. The PBPK model accurately simulated BEC in rats and mice at all exposure levels, as well as BEC reported in human males in previously published studies. Simulated peak BEC in human males following exposure to 50 and 600 ppm ranged from 7 to 23 microM and 86 and 293 microM, respectively. These results illustrate that inhalation of ethanol at or above the concentrations expected to occur upon refueling results in minimal BEC and are unlikely to result in toxicity. PMID- 9221834 TI - Cellular mechanisms for developmental toxicity of chlorpyrifos: targeting the adenylyl cyclase signaling cascade. AB - Developmental neurotoxicity caused by chlorpyrifos exposure is generally thought to target cholinesterase but chlorpyrifos may also act on cellular intermediates, such as adenylyl cyclase, that serve global functions in the coordination of cell development. In the current study, neonatal rats were exposed to apparently subtoxic doses of chlorpyrifos (no weight loss, no mortality) either on Postnatal Days 1-4 or on Postnatal Days 11-14, and the effects on components of the adenylyl cyclase cascade were evaluated in brain regions that are enriched (forebrain) or sparse (cerebellum) in cholinergic innervation, as well as in a nonneural tissue (heart). In all three, chlorpyrifos evoked deficits in multiple components of the adenylyl cyclase cascade: expression and activity of adenylyl cyclase itself, functioning of G-proteins that link neurotransmitter and hormone receptors to cyclase activity, and expression of neurotransmitter receptors that act through this cascade. Disruption of signaling function was not restricted to transduction of cholinergic signals but rather extended to adrenergic signals as well. In most cases, the adverse effects were not evident during the immediate period of chlorpyrifos administration, but appeared after a delay of several days. These results suggest that chlorpyrifos can affect cell development by altering the activity and reactivity of the adenylyl cyclase signaling cascade, a major control point for trophic regulation of cell differentiation. The effects are not restricted to cholinergic targets, nor even to the central nervous system. Hence, disruption of cell development by chlorpyrifos is likely to be more widespread than previously thought. PMID- 9221835 TI - Renal cell regeneration following oxidant exposure: inhibition by TGF-beta1 and stimulation by ascorbic acid. AB - Renal proximal tubular cell (RPTC) monolayers exposed to the model oxidant tert butylhydroperoxide (TBHP; 0.8 mM) for 1.5 hrs were 33 and 31% confluent after 1 and 4 days, respectively. Control monolayers remained 100% confluent throughout the experiment. Exogenous TGF-beta1 promoted monolayer deterioration by potentiating cellular death and suppressed EGF-stimulated regeneration of the RPTC monolayer. Net TGF-beta1 production in injured RPTC increased 1.7- and 3.2 fold on Days 1 and 2, respectively, and returned to control levels 4 days following TBHP treatment. An anti-TGF-beta antibody increased monolayer confluence to 50% and DNA content 1.3-fold 4 days after TBHP exposure. L-Ascorbic acid 2-phosphate (AscP) present only during the recovery period increased monolayer confluence to 67% but had no effect on RPTC proliferation, suggesting that AscP promoted monolayer regeneration by cellular migration/spreading. AscP present continuously had no effect on the extent of TBHP-induced injury but promoted regeneration of RPTC with increased monolayer confluence (1.8-fold) and DNA content (1.8-fold) and decreased cellular lysis by 52% 4 days following TBHP exposure. The results demonstrate that TBHP-induced injury increases net TGF beta1 production in RPTC and that autocrine TGF-beta1 inhibits regeneration of the monolayer by potentiating cellular injury and monolayer deterioration. The data also show that AscP is not cytoprotective during TBHP exposure but promotes RPTC regeneration by stimulating proliferation and migration/spreading and decreasing cellular death during the recovery period. PMID- 9221836 TI - The immunomodulatory effects of the herbicide propanil on murine macrophage interleukin-6 and tumor necrosis factor-alpha production. AB - Intraperitoneal (i.p.) exposure to propanil (3,4-dichloropropionanilide) has previously been shown to affect macrophage cytotoxicity. In this study, we compared the immunotoxic effects of propanil, after different routes of in vivo administration, on cytokine production by thioglycollate-elicited peritoneal macrophages. C57B1/6 mice were treated with either vehicle or 200 mg/kg propanil i.p., or with vehicle, 40, or 400 mg/kg propanil orally. Three or 7 days later, ex vivo production of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF alpha) by macrophages after lipopolysaccharide (LPS) stimulation was determined. Both oral and i.p. propanil exposure resulted in up to a 60-70% reduction in IL-6 and TNF-alpha production by the LPS-stimulated macrophages, depending on the route, postexposure time, and dose of propanil administered. Oral exposure to propanil also caused splenomegaly and thymic atrophy in animals in much the same manner as animals exposed via the i.p. route. In vitro exposure to propanil also significantly reduced macrophage cytokine production. Thioglycollate-elicited macrophages from normal mice were cultured in the continuous presence of 0, 10, or 20 microM propanil plus LPS. This exposure caused a significant reduction in IL-6 and TNF protein production by these macrophages in a concentration-dependent manner. Northern blot analysis demonstrated that the message levels of these cytokines were reduced by approximately the same percentage as the protein levels in propanil-treated macrophages, indicating a possible transcriptional or pretranscriptional target(s) for propanil. PMID- 9221837 TI - A physiologically based pharmacokinetic and pharmacodynamic model for paraoxon in rainbow trout. AB - Trout were exposed to an aqueous solution of 75 ng/ml paraoxon for 5 days at 12 degrees C. The relationships among paraoxon concentration in water and target organs, AChE inhibition, and carboxylesterase (CaE) detoxification of paraoxon were characterized quantitatively by development of a PBPK-PD model. The PKPD model structure consisted of brain, heart, liver, kidney, and remainder of the body, which were interconnected by blood circulation. The paraoxon tissue/blood partition coefficients were: plasma/water, 1.46; liver/plasma, 5.89; brain/plasma, 3.90; heart/plasma, 2.91; kidney/plasma, 0.45; and blood/plasma, 0.91. Turnover of AChE was characterized from a dose-response study, in which its zero-order synthesis rate and first-order degradation rate constant were determined in several tissues; for brain they were 7.67 pmol/min and 7.31 x 10( 5) hr(-1). The uptake and depuration clearances of paraoxon (Cl(u) = 0.651 and Cl(d) = 0.468 ml min(-1) g body wt(-1)) were determined using a compartmental model. During continuous water exposure to paraoxon, AChE activity in the tissues declined to new steady state values that were maintained by the synthesis of new AChE. CaE was shown by simulation to be an important pathway for detoxification of paraoxon. PMID- 9221838 TI - The role of intracellular calcium in antimony-induced toxicity in cultured cardiac myocytes. AB - Trivalent antimony, delivered as potassium antimonyl tartrate (PAT), has been previously shown to induce an oxidative stress and toxicity in cultured neonatal rat cardiac myocytes. The present study investigates the effect of PAT on intracellular free calcium ([Ca2+]i), which has been implicated in the toxicity of agents inducing oxidative stress, and explores its role in PAT toxicity. Exposure to 50 or 200 microM PAT led to progressive elevation in diastolic or resting [Ca2+]i and eventually a complete loss of [Ca2+]i transients that occurred well before cell death as assessed by LDH release. Prior loading of myocytes with the intracellular calcium chelator BAPTA (10 to 40 microM), protected against PAT toxicity in the presence and absence of extracellular calcium, and demonstrated a crucial role for [Ca2+]i in PAT toxicity. Exposure to 200 microM PAT in the absence of extracellular calcium slightly elevated [Ca2+]i, but only to levels comparable to resting [Ca2+]i for cells in 1.8 mM extracellular calcium. This demonstrated that although PAT toxicity was dependent on [Ca2+]i, a large increase above resting levels was not needed, and also that some calcium was mobilized from intracellular stores. However, the caffeine releasable pool of sarcoplasmic reticulum calcium was increased, not depleted, by exposure to 200 microM PAT. These results demonstrate that PAT disrupts [Ca2+]i handling and support a role for a calcium-dependent event, but do not support the necessity of events in PAT-induced cell death that are mediated by a large elevation in [Ca2+]i. PMID- 9221839 TI - Regioselective binding of 2,5-hexanedione to high-molecular-weight rat neurofilament proteins in vitro. AB - Previous studies have shown selective binding of the neurotoxicant 2,5 hexanedione (2,5-HD) to carboxyl-terminal domains of rat neurofilament (NF) M and H proteins in vitro. The present study was designed to further localize this binding in native rat NF preparations exposed to [14C]2,5-HD. Purified M and H proteins from 2,5-HD-treated NFs were subjected to cyanogen bromide (CNBr) cleavage, and the resultant peptides were separated by Tris-tricine SDS-PAGE and electroblotted to PVDF membranes. Peptides were identified by direct sequencing of stained bands and the relative radiolabeling of each peptide was determined by comparing band intensities in fluorographed blots. For NF-M, the highest label was found in CNBr 10, a peptide corresponding to residues 678-846 at the extreme carboxyl terminus. This region of the protein includes three highly conserved lysine-containing sequences believed to be critical to its function. For NF-H, the greatest binding was localized in CNBr 7 + 8, representing an incomplete cleavage product of residues 390-810. This peptide contains essentially all of the phosphorylation sites in the carboxyl terminus of NF-H, a domain believed to control NF interactions in the axon. Only minor radiolabeling was observed in other M or H peptides. Extensive dephosphorylation of NFs prior to 2,5-HD exposure had no effect on relative adduct levels in each protein. These results provide additional support for limited and specific binding of 2,5-HD to neurofilaments and indicate that the phosphorylation state of the protein may not substantially influence this binding. PMID- 9221840 TI - Modulation of T-helper cell populations: potential mechanisms of respiratory hypersensitivity and immune suppression. AB - Information presented at this symposium indicates that modulation of Th cell responses is one means by which xenobiotics may cause immunotoxicity. A shift from Th1 to Th2 responses can enhance both infectious and allergic disease. Hence, in some cases, a common mechanism may be responsible for effects that are generally considered to be very different. Because cytokines produced in the inflammatory process play a role in modulation of Th cell responses, there is a mechanism by which agents that appear to have only local effects at the portal of entry may, in fact, affect immune responses systemically. An understanding of conditions which trigger certain cytokine responses may be useful not only in understanding inflammation but also in predicting certain kinds of immunosuppressive and allergic responses. Future studies in this area are likely to provide insights into many areas of immunotoxicology. PMID- 9221846 TI - Abdominoperineal excision of the rectum--an endangered operation. Norman Nigro Lectureship. AB - PURPOSE: This study was undertaken to test the efficacy of an extreme policy of sphincter conservation by combining precise total mesorectal excision with low stapling techniques and endoluminal lavage to guard against implantation. METHODS: A total of 136 consecutive operations for cancer below 5 cm from the anal verge has been prospectively documented and followed for a mean of 7.7 (range, 1-18) years. A total of 105 of the operations were anterior resections (77 percent), and 31 were abdominoperineal excisions (23 percent). RESULTS: The oncologic results in the 105 patients who underwent anterior resections appear greatly superior to those of the patients who underwent abdominoperineal excisions, although the number of abdominoperineal excisions was small (31). Actuarial local recurrence at six years for anterior resection and total mesorectal excision was 1 percent for 85 curative procedures and 4 percent for all cases (n = 100), compared with 33 and 47 percent for abdominoperineal excisions (n = 15 and 31). Only four recurrences were observed below the level of the levators, three in the wound of an abdominoperineal excision and one in a stapled anastomosis after a palliative excision. No cases of nodal metastasis in the ischiorectal fossa were observed. CONCLUSION: In a unit specializing in sphincter conservation, precise total mesorectal excision from above appears oncologically superior to abdominoperineal excision. Three-fourths of patients with carcinoma of the lower one-third of the rectum can be offered sphincter conserving surgery, although temporary defunctioning is probably prudent in such cases. The wound of an abdominoperineal excision may be a prerequisite for perineal recurrence, which may often be caused by implantation. PMID- 9221847 TI - Vaginal fistula following restorative proctocolectomy. AB - Vaginal fistula (VF) is a devastating complication following restorative proctocolectomy. PURPOSE: This study was designed to examine the perioperative factors influencing the outcome and management of vaginal fistula. METHOD: Between October 1983 and September 1994, 526 women underwent restorative proctocolectomy. Nineteen develop VF (3.6 percent), and six were referred from other institutions with this complication. These 25 women were followed for a minimum of nine months. RESULTS: Preoperative diagnosis of ulcerative colitis was made in 23 of the patients with VF (92 percent), and indeterminate colitis and familial adenomatous polyposis was determined in the rest of the patients. Postoperatively, 12 of the 23 women (52 percent) with a preoperative diagnosis of ulcerative colitis had clinical/pathologic findings of Crohn's disease, and 1 woman was reclassified as having indeterminate colitis. Postoperative pelvic sepsis was significantly higher in women with VF than in those without VF (26.3 vs. 6.3 percent; P = 0.003). Median time until occurrence of VF following loop ileostomy closure was later for women with delayed findings of Crohn's disease at 16.5 (range, <1-72) months, compared with women without Crohn's disease at 0.5 (range, <1-67) months (P < 0.05). Of the 163 women with handsewn anastomosis performed at our institution, 12 developed VF (7.4 percent). In contrast, 7 of the 363 patients with stapled anastomosis had VF (1.9 percent; P = 0.003). Site of VF was found at the anastomosis in 12 patients, below in 12 patients, and above in 1 patient. Presence of Crohn's disease and anastomotic technique did not influence the site of VF. Initial management of VF consisted of transanal repair in 20 patients (advancement flap, 12; direct repair, 6; and neoileoanal anastomosis, 2), seton in 1 patient, transabdominal approach in 1 patient, transvaginal in 1 patient, observation in 1 patient, and pouch excision in 1 patient. Of the 13 women without Crohn's disease, 12 had transanal repair (10 healed, 1 had recurrence, and 1 had pouch excision), and 1 had successfully repair with transabdominal technique, for an overall success rate of 84.6 percent. Of the 12 women with VF and delayed findings of Crohn's disease, transanal repair was performed on 9, 1 had pouch excision without repair, 1 had seton placement and pouch excision, and 1 underwent observation. Transanal technique of repair in women with Crohn's disease successfully healed three women (33.3 percent). Overall, of the 12 women with delayed findings of Crohn's disease, 6 had pouch excision, 3 had recurrences, and 3 healed. CONCLUSION: VF is an uncommon complication following restorative proctocolectomy and is associated with a high incidence of pelvic sepsis and handsewn anastomosis. Late presentation of VF is more common with Crohn's disease and is associated with a poor prognosis and pouch salvage rate. Transanal techniques are an effective means of VF repair. PMID- 9221849 TI - Functional results after restorative proctocolectomy complicated by pouchitis. AB - This study aimed to examine the incidence and cumulative risk of pouchitis after restorative proctocolectomy for ulcerative colitis and to evaluate the clinical and functional results in patients with pouchitis. METHODS: A total of 291 patients had proctocolectomy with ileal pouch-anal anastomosis for ulcerative colitis between January 1985 and January 1996. During follow-up, 65 patients had one or more episodes of pouchitis based on clinical, histologic, and endoscopic criteria. Functional results and patient satisfaction in these patients were compared with those of 65 matched control patients who had experienced no episodes of pouchitis. RESULTS: Pouchitis developed in 65 patients (22 percent), giving rise to a cumulative frequency of 28 percent at 11 years after the operation. Only 13 patients (4.5 percent) had chronic pouchitis that required long-lasting treatment. A permanent ileostomy had to be constructed in one patient (0.3 percent) because of pouchitis. During the last year (1995), 60 percent of patients with pouchitis had medication, most often metronidazole and/or corticosteroids. Defecation frequency per 24 hours was 6.7 for all patients with pouchitis, 8.2 for those with chronic pouchitis (P < 0.05), and 6.3 for patients without pouchitis. Nighttime defecation occurred in 44 (80 percent) patients with pouchitis, compared with 37 (67 percent) of those without pouchitis (P > 0.05). Frequencies of soiling or flatus incontinence did not differ between the two groups. During the last year, 43 (80 percent) of the pouchitis patients, who answered the questionnaire, were working all the year or were on sick-leave less than one month. CONCLUSIONS: Episodic pouchitis is easily treated and causes minimum functional consequences, whereas chronic pouchitis increases defecation frequency and needs prolonged medication. Pouchitis seems not to be a major threat to preventing the use of restorative proctocolectomy in ulcerative colitis, but still the small group of chronic pouchitis patients remains a problem. PMID- 9221848 TI - Treatment of colorectal and ileoanal anastomotic sinuses. AB - PURPOSE: This study is designed to describe a technique and report results for treating low anastomotic sinuses. METHODS: Restorative proctocolectomy and complicated low anterior resections were protected with diverting loop ileostomy. Contrast enemas identified anastomotic problems before ileostomy closure. Pouch anal or colorectal anastomotic sinuses that failed to resolve with observation were treated before intestinal continuity was restored. With the patient receiving regional or general anesthesia, a rigid proctoscope or anoscope was used to identify the sinus opening. The common wall between the sinus and the bowel lumen was divided under direct vision with laparoscopic cautery scissors, and the sinus cavity was debrided with a suction cautery wand placed through the scope. RESULTS: Six patients with anastomotic sinuses have received outpatient treatment in the described manner during the past two years. Four patients had restorative proctocolectomies for ulcerative colitis, and two had low anastomosis for rectal cancer. Three patients presented with pelvic sepsis before the contrast study; the remainder were asymptomatic. Division of anastomotic sinus was performed one to eight months after diagnosis of the sinus. Following division, anastomotic cavities resolved in five patients by 1 month and in one patient by 12 months. In these six patients, there was one dilatable anastomotic stricture but no other anastomotic complications at follow-up 5 to 16 (mean, 9.2) months after sinus division. CONCLUSION: When used in conjunction with fecal diversion, sinus unroofing by division of the common wall between the sinus and bowel lumen treats low pelvic sinuses. PMID- 9221851 TI - Early postoperative results of a prospective series of laparoscopic vs. Open anterior resections for rectosigmoid cancers. AB - PURPOSE: This study was undertaken to compare postoperatively laparoscopic (LAR) with open (OAR) anterior resection in patients with rectosigmoid cancers. METHODS: Forty consecutive patients were divided into two groups: 20 patients (9 males) were allocated to LAR and 20 patients (6 males) to OAR. RESULTS: Median age in the LAR group was 62 (range, 39-77) years, and in the OAR group, it was 61 (range, 43-84) years (P = 0.9). Median lengths of the distal margin of clearance beyond the tumor were 4 (range, 2-8) cm and 4.5 (range, 3-7.5) cm in the LAR and OAR groups, respectively (P = 0.35). Median numbers of lymph nodes harvested were 20 (range, 7-49) and 19 (range, 7-97) for the LAR and OAR groups, respectively (P = 0.44). Median operating times were 90 (range, 55-185) minutes and 73 (range, 40 140) minutes in the LAR and OAR groups, respectively (P = 0.08). Blood losses were 50 (range, 50-800) ml and 50 (range, 50-1,500) ml in the LAR and OAR groups, respectively. There was no intraoperative complication in either group, and no laparoscopic patient was converted to an open procedure. Median length of extraction site incision in the LAR group was 5.5 (4-13) cm, and length of incision in the OAR group was 18 (8-25) cm (P < 0.002). CONCLUSION: There were no significant differences between the two groups with regard to duration of parenteral analgesia, starting of fluid and solid diet after surgery, or time to first bowel movement and time to discharge from the hospital. PMID- 9221852 TI - Relationship between fecal sampling times and sensitivity and specificity of immunochemical fecal occult blood tests for colorectal cancer: a comparative study. AB - PURPOSE: The present study was conducted to assess the accuracy of three testing methods using an immunochemical fecal occult blood test based on the number of samples as the optimum means for screening of colorectal cancer. METHOD: One hundred eighty-four patients with colorectal cancer and 368 healthy controls served as the subjects for this study. Each subject was tested by an immunochemical fecal occult blood test for three consecutive days. For evaluation of the most desirable number of sampling times, we used the results of the first day for the one-day method, results of the first and second days for the two-day method, and results of three consecutive days for the three-day method. Sensitivities and specificities of the three testing methods were evaluated. RESULTS: Sensitivities of an immunochemical fecal occult blood test were calculated as 67.9 percent for the one-day method, 88 percent for the two-day method, and 90.8 percent for the three-day method; specificity was as follows: 97.5 percent for the one-day method, 95.6 percent for the two-day method, and 92.1 percent for the three-day method. A significant difference in sensitivity was shown between the one-day and two-day and the one-day and three-day methods (P < 0.01); also, a significant difference in specificity was shown between one day and three-day and two-day and three-day methods (P <0.05). CONCLUSIONS: These results indicate that the two-day method is recommended for immunochemical fecal occult blood testing as a means of screening for colorectal cancer. PMID- 9221850 TI - Selective chemoembolization in the management of hepatic metastases in refractory colorectal carcinoma: a phase II trial. AB - PURPOSE: Metastatic involvement of the liver frequently determines the evolution of the clinical picture in colorectal cancer patients. We examined the efficacy and toxicity of chemoembolization in this setting, identifying prognostic factors to define patients most likely to benefit from the procedure. METHODS: Forty patients underwent chemoembolization of metastatic liver lesions from colorectal carcinoma. Selective angiography of the hepatic artery was performed to identify the feeding vessels of the metastatic lesions. The injected chemoemulsion consisted of 1,000 mg of 5-fluorouracil, 10 mg of mitomycin C, and 10 ml of ethiodized oil in a total volume of 30 ml. Gelfoam embolization then followed, until stagnation of blood flow was achieved. Patients were evaluated for response, overall survival, and toxicities. RESULTS: Overall median survival from date of first chemoembolization was ten months. Factors that predicted a longer median survival included favorable performance status (24 months), serum alkaline phosphatase and lactate dehydrogenase levels less than three times normal (24 and 12 months, respectively), and metastatic disease confined to the liver (14 months). Most patients tolerated the procedure well. The most common side effects were transient fevers, abdominal pain, and fatigue. Three patients died within one month from the procedure. CONCLUSION: This study suggests that chemoembolization of hepatic metastases in colorectal cancer should be further evaluated; it may be beneficial in patients who have failed systemic chemotherapy, have a good performance status, and have metastatic disease confined to the liver. PMID- 9221853 TI - Coexpression of Bcl-2, c-Myc, and p53 oncoproteins as prognostic discriminants in patients with colorectal carcinoma. AB - PURPOSE: This study was undertaken to evaluate the clinical use of Bcl-2, c-Myc, and p53 oncoproteins, either singly or in combination, as prognostic discriminants relative to recurrence and overall survival in patients with Dukes B or C colorectal carcinoma. METHODS: Analyses were made on archival pathology tissues of 48 patients with colorectal cancer. The oncoproteins were localized using commercially available monoclonal antibodies: clone 124 for Bcl-2, 9E11 for c-Myc, and DO-7 for p53. The avidin-biotin peroxidase complex method was used. Patients were followed up for a period of 2 to 36 months. RESULTS: Expression of Bcl-2 and c-Myc was cytoplasmic, whereas nuclear p53 immunoreactivity was localized in the tumor cells. Sixty percent (29/48), 65 percent (31/48), and 37 percent (18/48) of the tumors showed overexpression of Bcl-2, c-Myc, and p53 oncoproteins, respectively. Fifty-four percent (18/33) and 100 percent (9/9) of moderately and poorly differentiated tumors, respectively, were positive for Bcl 2 (P < 0.01). No such correlation was noted for c-Myc and p53 oncoproteins. Univariate analysis showed that patients with Bcl-2 and c-Myc overexpression were associated with poorer overall survival than patients with Bcl-2-negative (P < 0.0124) and c-Myc-negative (P < 0.036) tumors. In addition, when patients were subgrouped according to Dukes stage, a statistically significant poorer overall survival was observed in Dukes C patients with Bcl-2-positive tumors (P < 0.017). Furthermore, multivariate analysis revealed that coexpression of three oncoproteins was predictive of a worse prognosis than for those individuals expressing none of the oncoproteins (P < 0.031) and only one positive oncoprotein (P < 0.014). CONCLUSION: These findings suggest that oncoprotein coexpression possesses significant prognostic and potential therapeutic value; incorporation of molecular markers into future prospective randomized trials is advisable. PMID- 9221854 TI - Possible effect of pneumoperitoneum on the spreading of colon cancer tumor cells. AB - PURPOSE: By using a murine hepatic metastatic model, we tried to investigate the possible influence of gas insufflation in colon cancer cells spreading from the portal system to the liver. METHODS: After transducing the human placental ALP gene into murine colon cancer cell line CT26, we successfully selected a clone of CT26/DAP that would yield a specific color following histochemical staining. Fifty mice were assigned into two groups, receiving either an intrasplenic injection of 10(6) CT26/DAP cells alone or the cells followed by intra-abdominal helium insufflation with the pressure of 15 cm H2O for ten minutes. Five mice in each group were used to observe their survival and the other mice were killed at four different time periods: 10 minutes, 24 hours, 48 hours, and 72 hours following cell injection. The livers and spleens were removed for histochemical staining. By counting the numbers of specific dark reddish spots of CT26/DAP cells, we could estimate the number of tumor cells on the hepatic surface. RESULTS: At the very beginning following tumor cell injection, we found a significantly greater number of tumor cells on the hepatic surface in mice with gas insufflation (6354 +/- 1072 vs. 2133 +/- 223, respectively; P = 0.012). But the difference of these two groups became smaller and smaller as time went by. The number of tumor cells on the hepatic surface would reach the lowest level at postoperative 48 hours, and the tumor foci then began to grow both in size and number. The above patterns of dynamic change in tumor cell distribution were similar in mice both with and without gas insufflation. Average survival was slightly shorter in mice with gas insufflation, but the difference was not statistically significant. CONCLUSION: Pneumoperitoneum caused by gas insufflation may increase tumor cell spread from the portal system to the liver at the very beginning stage; however, there was no significant difference in long term survival between mice with and without gas insufflation in this murine animal model. PMID- 9221855 TI - Chemotherapy for desmoid tumors in association with familial adenomatous polyposis. AB - PURPOSE: This study was designed to assess the effect of chemotherapy on complex desmoid tumors associated with familial adenomatous polyposis. METHODS: Five patients (3 males, 2 females; age range, 29-45 years) had symptomatic, unresectable intra-abdominal desmoid tumors in association with familial adenomatous polyposis that were unresponsive to conventional medical therapy. Each patient was treated with a cytotoxic chemotherapeutic regimen consisting of doxorubicin and dacarbazine followed by carboplatin and dacarbazine. Response to treatment was assessed by measurement of tumor size using computerized tomography. Follow-up has been for a mean of 22 (range, 10-30) months. RESULTS: One patient has had a complete response, and three patients have had a partial response, with a reduction in tumor volume of at least 50 percent. One patient had a minimum response to treatment and developed a rapid increase in tumor size on cessation of therapy. Complications of treatment included febrile neutropenia, severe epistaxis, and subclavian vein thrombosis. CONCLUSIONS: The cytotoxic chemotherapeutic regimen described is effective in the treatment of selected unresectable desmoid tumors associated with familial adenomatous polyposis and should be considered in symptomatic patients who do not respond to conventional medical therapy. PMID- 9221856 TI - Clinical value of colonic irrigation in patients with continence disturbances. AB - Continence disturbances, especially fecal soiling, are difficult to treat. Irrigation of the distal part of the large bowel might be considered as a nonsurgical alternative for patients with impaired continence. PURPOSE: This study is aimed at evaluating the clinical value of colonic irrigation. METHODS: Thirty-two patients (16 females; median age, 47 (range, 23-72) years) were offered colonic irrigation on an ambulatory basis. Sixteen patients suffered from fecal soiling (Group I), whereas the other 16 patients were treated for fecal incontinence (Group II). Patients were instructed by enterostomal therapists how to use a conventional colostomy irrigation set to obtain sufficient irrigation of the distal part of their large bowel. Patients with continence disturbances during the daytime were instructed to introduce 500 to 1,000 ml of warm (38 degrees C) water within 5 to 10 minutes after they passed their first stool. In addition, they were advised to wait until the urge to defecate was felt. Patients with soiling during overnight sleep were advised to irrigate during the evening. To determine clinical outcome, a detailed questionnaire was used. RESULTS: Median duration of follow-up was 18 months. Ten patients discontinued irrigation within the first month of treatment. Symptoms resolved completely in two patients. They believed that there was no need to continue treatment any longer. Irrigation had no effect in two patients. Despite the fact that symptoms resolved, six patients discontinued treatment because they experienced pain (n = 2) or they considered the irrigation to be too time-consuming (n = 4). Twenty-two patients are still performing irrigations. Most patients irrigated the colon in the morning after the first stool was passed. Time needed for washout varied between 10 and 90 minutes. Frequency of irrigations varied from two times per day to two times per week. In Group I, irrigation was found to be beneficial in 92 percent of patients, whereas 60 percent of patients in Group II considered the treatment as a major improvement to the quality of their lives. If patients who discontinued treatment because of washout-related problems are included in the assessment of final outcome, the success rate is 79 and 38 percent respectively. CONCLUSIONS: Patients with fecal soiling benefit more from colonic irrigation than patients with incontinence for liquid or solid stools. If creation of a stoma is considered, especially in patients with intractable and disabling soiling, it might be worthwhile to treat these patients first by colonic irrigation. PMID- 9221857 TI - "Ideal" length of stay after colectomy: whose ideal? AB - PURPOSE: In response to external pressure to achieve an idealized length of stay after colon resection, a study was designed to define perioperative factors that significantly impact average length of stay (ALOS). METHODS: We retrospectively reviewed the records of 226 patients undergoing open colon resection from 1988 to 1995 to determine the effects of age, type of procedure, nature of the procedure (elective vs. emergency), and postoperative course on ALOS. Statistics were calculated by Student's t-test, chi-squared analysis, and analysis of variance. RESULTS: Average length of stay was 10 (range, 4-34) days, with a significant trend toward lower ALOS in recent years; ALOS in 1988 averaged 11 days, whereas in 1994, ALOS averaged 9 days (r2 = 0.118; P < 0.001). Patients younger than 65 years of age had an ALOS of 9 days vs. 11 days in patients older than 65 years (P = 0.0024). Patients with anastomoses on the right and left side had similar ALOS (8.5 vs. 9.1 days), whereas creation of a stoma was associated with a significantly higher ALOS (12.1 days; P < 0.00001). The need for postoperative nasogastric intubation (14.9 vs. 9.3 days) and the performance of emergency operations (12.2 vs. 6.5 days) were also associated with a significantly higher ALOS (P < 0.00001). CONCLUSIONS: Caution must be exercised in accepting rigid criteria for length of stay for patients undergoing colorectal resections, as uncontrollable clinical variables are involved in defining the "ideal" patient. PMID- 9221858 TI - Anal sensitivity test: what does it measure and do we need it? Cause or derivative of anorectal complaints. AB - PURPOSE: This study was undertaken to determine the anal sensitivity in controls and in different patient groups and to establish factors that determine anal sensitivity. METHODS: Anorectal function tests were performed in 387 patients with different anorectal diseases. Anal sensitivity was measured in 36 controls. Anal sensitivity was measured by means of mucosal electrosensitivity (MES) using a catheter with two electrodes placed in the anal canal. A constant current (square wave stimuli 100 microsec, pulses per second) was increased stepwise from 1 to 20 mAmp until the threshold sensation was reached. Other tests used were anal manometry (maximum basal pressure, maximum squeeze pressure, rectal compliance (maximum rectal volume and pressure), endosonography (submucosal thickness), defects and thickness of internal and external sphincter), electromyography (maximum contraction pattern, Grade 1 (solitary contractions) to Grade 4 (interference pattern)), and pudendal nerve terminal motor latency. Multiple regression analysis was performed. It was postulated that age, local conditions (anal scars, anal fissures, hemorrhoids, mucosal prolapse, proctitis, sphincter thickness and defects, and submucosal thickness), and neurologic factors could influence anal sensitivity. RESULTS: Controls had an MES of 3.4 +/- 1.7. MES was significantly increased compared with controls in patients with fecal incontinence, soiling, hemorrhoids, mucosal prolapse, constipation, anal scars, anal surgery, and sphincter defects; patients with fecal incontinence had the highest MES (6.7 +/- 4.3; P < 0.0001). Patients with anal fissures and proctitis showed no differences compared with controls. MES correlated significantly with age (R = 0.29), maximum basal pressure (R = -0.29), maximum squeeze pressure (R = -0.32), submucosal thickness (R = 0.19), maximum contraction pattern (R = -0.39), single-fiber electromyography (R = 0.39), and maximum rectal volume and pressure (0.14). Multiple regression analysis showed that age, internal sphincter defects, and submucosal thickness significantly influenced anal sensitivity, but explained only 10 percent of the variance. CONCLUSION: Anal sensitivity is diminished in all patients with anorectal diseases except for anal fissures and proctitis. There are correlations with other anorectal function tests. Anal sensitivity is determined for 10 percent by age, internal sphincter defects, and thickness of the submucosa. Anal sensitivity measurement, therefore, has limited clinical value and should be used in conjunction with other tests in a research setting. PMID- 9221859 TI - Internal rectal intussusception seldom develops into total rectal prolapse. AB - PURPOSE: This study was designed to analyze how often internal rectal intussusception develops into total rectal prolapse. METHODS: Repeated investigations with defecography were performed in 312 patients because of persisting symptoms. In 79 patients who had a rectal intussusception at the first defecography, results of the second defecography and the patients' records were studied. RESULTS: A total of 38 patients had not undergone any surgical treatment of rectal intussusception or rectal prolapse between the first and second defecographies. One of these patients had a rectal prolapse at the second defecography, and another developed a clinical prolapse after the second defecography. CONCLUSIONS: The present study demonstrates that the risk of developing a rectal prolapse in patients with rectal intussusception is small. This risk should, therefore, not be used as an indication for surgery. PMID- 9221860 TI - Prospective trial of pelvic floor retraining in patients with fecal incontinence. AB - PURPOSE: Our aim was to prospectively evaluate pelvic floor retraining (PFR) in improving symptomatic fecal incontinence. METHODS: PFR was used to treat 30 patients with fecal incontinence (28 women; age range, 29-85 (median, 68) years). PFR was performed by a physiotherapist in the outpatient department according to a strict protocol and included biofeedback using an anal plug electromyometer. Manometry (24 patients), pudendal nerve terminal motor latency (PNTML, 16 patients), and anal ultrasound (14 patients) were done before commencing therapy. Independent assessment of symptoms was done at the commencement of therapy, at 6 weeks, and at 6 and 12 months posttherapy. RESULTS: Twenty patients (67 percent) had improved incontinence scores, with eight patients (27 percent) being completely or nearly free of symptoms. Of 28 patients followed up longer than six months, 14 achieved a 25 percent or greater improvement at six weeks, which was sustained in all cases. Fourteen had an initial improvement of less than 25 percent, with only four (29 percent) showing later improvement (P < 0.0001). There was no relationship between results of the therapy and patient age, initial severity of symptoms, etiology of incontinence, and results of anal manometry, PNTML, and anal ultrasound. CONCLUSIONS: PFR is a physical therapy that should be considered as the initial treatment in patients with fecal incontinence. An improvement can be expected in up to 67 percent of patients. Initial good results can predict overall outcome. PMID- 9221861 TI - Biofeedback in colorectal practice: a multicenter, statewide, three-year experience. AB - PURPOSE: Biofeedback treatment is often offered to patients in colorectal centers; however, standards of treatment are still lacking. A dedicated team approach is desirable but difficult to coordinate. We present our three-year experience of electromyographic-based biofeedback treatment offered within a multicenter, statewide organization. METHODS: Between October 1992 and October 1995, 188 patients completed a biofeedback treatment program in one of five coordinated centers within a 200-mile radius. A unified common database was established and continuously updated. A colorectal surgeon served as statewide director, and dedicated teams were established at each location. Each local team included the medical director and a certified biofeedback therapist and had access to a dietitian and a nurse data coordinator. Electromyographic-based biofeedback sessions were given weekly, and a home trainer program was established. RESULTS: A total of 116 patients with chronic constipation had a mean of eight (range, 2-14) weekly sessions. A total of 72 patients with fecal incontinence had a mean of seven (range, 2-11) weekly sessions. A total of 84 percent of the constipated and 85 percent of the incontinent patients had significant improvement with biofeedback treatment. Patient compliance and satisfaction were high. Constipated patients increased the mean number of weekly unassisted bowel movements from 0.8 to 6.5. Incontinent patients decreased the mean number of weekly gross incontinence episodes from 11.8 to 2. CONCLUSIONS: Biofeedback treatment can be extremely successful in both incontinent and constipated patients. A large geographic area can be covered with coordinated centers in which each dedicated team uses a unified treatment protocol, and a common database is established. PMID- 9221862 TI - Management of recurrent rectal prolapse. AB - PURPOSE: Many operations have been described for the management of rectal prolapse. Despite an overall recurrence rate of greater than 15 percent, few reviews address how to deal with this problem. This report summarizes our experience with recurrent rectal prolapse and includes suggestions for reoperative management of failed repairs from both abdominal and perineal approaches. PATIENTS AND METHODS: Fourteen patients (3 male) ranging in age from 22 to 92 (mean, 68) years underwent operative correction of recurrent rectal prolapse. Average time from initial operation to recurrence was 14 (range, 6-60) months. Initial operations (before recurrence) were as follows: perineal proctectomy and levatorplasty (10), anal encirclement (2), Delorme's procedure (1), and anterior resection (1). Operative procedures performed for recurrence were as follows: perineal proctectomy and levatorplasty (7), sacral rectopexy (abdominal approach; 3), anterior resection with rectopexy (2), Delorme's procedure (1), and anal encirclement (1). Average length of follow-up was 50 (range, 9-115) months. RESULTS: No further episodes of complete rectal prolapse were observed during this period. Preoperatively, three patients were noted to be incontinent to the extent that necessitated the use of perineal pads. The reoperative procedures failed to restore fecal continence in any of these three individuals. One patient died in the postoperative period after anal encirclement from an unrelated cause. CONCLUSION: Surgical management of recurrent rectal prolapse can be expected to alleviate the prolapse, but not necessarily fecal incontinence. Perineal proctectomies can be safely repeated. Resectional procedures may result in an ischemic segment between two anastomoses, unless the surgeon can resect a previous anastomosis in the repeat procedure. Nonresectional procedures such as the Delorme's procedure should be strongly considered in the management of recurrent rectal prolapse if a resectional procedure was performed initially and failed. PMID- 9221863 TI - Long-term results of total pelvic floor repair for postobstetric fecal incontinence. AB - PURPOSE: This study was designed to assess the long-term results of total pelvic floor repair for postobstetric neuropathic fecal incontinence. METHOD: Sixty three of 75 women who had undergone total pelvic floor repair for postobstetric neuropathic fecal incontinence were traced and interviewed a median of 36 (18-78) months after surgery. Thirty-nine patients agreed to repeat anorectal physiology. RESULTS: Six patients required further surgery for persistent incontinence (colostomy, 4; graciloplasty, 2). For the remaining 57 patients, incontinence improved greatly in 28 (49 percent) patients, mildly in 13 (23 percent), and not at all in 16 (28 percent); daily incontinence was present in 41 patients (73 percent) before the operation but persisted in 13 (23 percent). Only eight (14 percent) patients were rendered completely continent; those with marked improvement were socially more active than those with little or no improvement. Resting and maximum squeeze pressures, anal canal sensation, rectal sensation, and pudendal nerve terminal motor latency did not predict outcome. Perineal descent, obesity, and a history of straining before the operation were all associated with a poor outcome. CONCLUSION: Total pelvic floor repair rarely renders patients with postobstetric neuropathic fecal incontinence completely continent but substantially improves continence and lifestyle in approximately one-half of them. The operation is less successful in obese patients and in those with a history of straining or perineal descent. PMID- 9221865 TI - Lower gastrointestinal bleeding. AB - BACKGROUND: Lower gastrointestinal bleeding can be a confusing clinical conundrum, the satisfactory evaluation and management of which requires a disciplined and orderly approach. Diagnosis and management has evolved with the development of new technology such as selective mesenteric angiography and colonoscopy. PURPOSE: This study was undertaken to review the available data in the literature and to determine the current optimum method of evaluation and management of lower gastrointestinal hemorrhage most likely to result in a successful outcome. METHODS: Data available on the topic of lower gastrointestinal bleeding in the English literature were obtained via MEDLINE search and were reviewed and analyzed. RESULTS: The colonic origin of lower gastrointestinal hemorrhage in order of decreasing incidence is diverticulosis, inflammatory bowel disease, including ischemic and infectious colitis, colonic neoplasia, benign anorectal disease, and arteriovenous malformations. Approximately 10 to 15 percent of all cases of rectal bleeding are attributable to a cause that is proximal to the ligament of Treitz. Small intestinal sources such as arteriovenous malformations, diverticula, and neoplasia account for between 3 and 5 percent of all cases. Colonoscopy successfully identified an origin in severe hematochezia in 74 to 82 percent of cases. Mesenteric angiography has a sensitivity of 42 to 86 percent. The best method of management depends on whether hemorrhage persists, the severity of continued hemorrhage, the cumulative transfusion requirement, and the specific origin of bleeding. CONCLUSION: Lower gastrointestinal hemorrhage is a complex clinical problem that requires disciplined and sophisticated evaluation for successful management. Diverticulosis is the most common cause. Colonoscopy is the diagnostic procedure of choice both for its accuracy in localization and its therapeutic capability. Selective mesenteric angiography should be reserved for those patients in whom colonoscopy is not practical. Precise identification of the bleeding source is crucial for a successful outcome. Specific directed therapy, such as segmental colonic resection for bleeding diverticulosis, is associated with the highest success rate and the lowest morbidity. A complete review of lower gastrointestinal bleeding is contained herein. PMID- 9221864 TI - Local nitroglycerin for treatment of anal fissures: an alternative to lateral sphincterotomy? AB - PURPOSE: Nitric oxide is an important neurotransmitter mediating internal anal sphincter relaxation. Patients suffering from fissure-in-ano were treated with topical nitroglycerine. The clinical evidence for therapeutic adequacy was examined in a prospective, randomized study. METHODS: The study included 35 patients with acute and chronic anal fissures. In Group A, including 20 patients with the clinical diagnosis of acute (12 patients) and chronic (8 patients) anal fissures, treatment consisted of topical nitroglycerine. Group B, consisting of 15 patients (10 acute and 5 chronic fissures), received topical anesthetic gel during therapy. Manometry was performed before and on days 14 and 28 in the course of topical application of either 0.2 percent glyceryl trinitrate ointment or anesthetic gel (lignocaine). Anal pressures were documented by recording the maximum resting and squeeze pressures. RESULTS: In 60 percent of cases treated with topical nitroglycerine (Group A, 11 acute (91.6 percent) and 1 chronic (12.5 percent)), anal fissure healed within 14 days, in contrast to Group B in which no healing was observed. The healing rate after one month was 80 percent (11 acute (91.6 percent); 5 chronic (62.5 percent)) in Group A and was significantly superior to Group B (healing rate, 40 percent: 5 acute (50 percent); 1 chronic (20 percent)). DISCUSSION: Previously increased maximum resting pressures decreased from a mean value of 110 to 87 cm H2O. This represents a mean reduction of 20 percent (P = 0.0022). We also noted a significant decrease in squeeze pressures (from 177.8 to 157.9 cm H2O (11 percent)). However, anal pressures did not decrease significantly in the four chronic fissure patients from Group A, whose fissures only healed after 28 days. Similarly to these Group A chronic fissure patients, no significant anal pressure reduction was observed in any Group B patients. Except for mild headache (20 percent), no side effects of treatment were reported. CONCLUSIONS: Topical application of nitroglycerine represents a new, easily handled, and effective alternative in the treatment of anal fissures. All of our patients reported a dramatic reduction in acute anal pain. However, it should be noted that a lack of sphincter tone reduction is a likely reason for the great tendency of chronic anal fissures to recur. PMID- 9221866 TI - Acute colonic surgery and unrecognized hypothyroidism: a warning. Report of six cases. AB - PURPOSE: This study was designed to highlight the significant morbidity related to undetected hypothyroidism in the elderly who are undergoing emergency surgery. METHOD: Case reports of six patients who presented with acute colonic surgical conditions are reviewed. RESULTS: Six cases of undetected hypothyroidism in a group of elderly patients was unmasked at the time of surgery for acute colonic conditions or in the perioperative period. These patients experienced increased morbidity, but once detected and treated, all but one had an uneventful recovery. CONCLUSION: Unrecognized hypothyroidism may lead to unnecessary surgery or even a potentially fatal outcome. A heightened awareness of this not so uncommon entity is mandatory. PMID- 9221867 TI - Neuronal intestinal dysplasia presenting as an abdominal mass: report of a case. AB - An unusual clinical presentation of a patient with neuronal intestinal dysplasia is presented. A 46-year-old male noted a palpable mass in the right lower quadrant of his abdomen for two months. A computed axial tomographic scan showed a thickened wall of the cecum with a tumor-like appearance. The excised specimen consisted of a mass caused by the thickened, edematous wall of the dilated cecum and appendix. The wall of the cecum and appendix measured up to 2.5 and 0.8 cm, respectively, in thickness. Microscopic studies showed extensive hyperplasia and hypertrophy of the ganglia and nerve plexuses and hypertrophy of the muscularis propria, consistent with neuronal intestinal dysplasia. PMID- 9221868 TI - The usefulness of carcinoembryonic antigen in postoperative colorectal cancer patients. PMID- 9221869 TI - Cecal ischemia mimicking carcinoma. PMID- 9221870 TI - Chiral separations by capillary zone electrophoresis: present state of the art. PMID- 9221871 TI - Monomeric and polymeric chiral surfactants as pseudo-stationary phases for chiral separations. PMID- 9221872 TI - Capillary zone electrophoresis at subzero temperatures II: chiral separation of biogenic amines. AB - The separation of enantiomer pairs of eight biogenic amines by capillary zone electrophoresis (CZE) was investigated with heptakis (2,6-di-O-methyl)-beta cyclodextrin as the chiral selector at temperatures ranging from -20 degrees to 40 degrees C by using a commercial electrophoresis unit retrofitted with an external thermostated refrigerated circulating bath in order to assist the original cooling system. Sodium phosphate in both neat aqueous and methanolic media at pH 2.5, as measured by the glass pH electrode, were used with the fused silica capillary from 1.5 degrees to 40 degrees C and -20 degrees to 40 degrees C, respectively. The effect of temperature on enantioselectivity was found to depend on the number of phenolic hydroxyl groups in the molecule. Upon lowering the temperature from 40 degrees to -20 degrees C, the chiral selectivity of the system, as measured by the relative mobility difference, increased tenfold for the amines with two vicinal phenolic hydroxyls, whereas the increase was insignificant for those having no phenolic hydroxyl groups. The complex formation constants of three amines which have the same molecular structure but the number of phenolic hydroxyl groups were determined at different temperatures and the thermodynamic parameters as well as compensation temperatures for the process were evaluated. Whereas the compensation temperature was 690 K for the amine without phenolic hydroxyl group, it was < 400 K for the amines with one or two phenolic hydroxyl groups. The difference in the compensation temperatures indicates that the intrinsic mechanisms of their complexation with the chiral selector are not the same; this may account for the discrepancies observed in the temperature dependency of the chiral selectivity. The enthalpy change per phenolic hydroxyl group was 2.5 kcal mol(-1), which compares favorably with the typical value for a single hydrogen bond. Therefore, when the amines have phenolic hydroxyl groups, the strong increase in chiral selectivity with decreasing temperature may be due to enhanced H-bonding between the cyclodextrin and the phenolic hydroxyls under the conditions employed in this study. PMID- 9221873 TI - Three approaches to enantiomer separation of beta-adrenergic antagonists by capillary electrochromatography. AB - Three different capillary electrochromatographic methods for the enantiomer separation of beta-adrenergic antagonists (acebutolol, alprenolol, atenolol, metoprolol, pindolol, prenalterol, and propranolol) were applied using different cyclodextrins (beta-cyclodextrin, carboxymethyl-beta-cyclodextrin and dimethyl beta-cyclodextrin) added to the electrolyte, a cross-linked protein-gel (cellobiohydrolase I) and a molecularly imprinted ((R)-enantiomer of propranolol) superporous polymer as chiral selectors. Through use of these different separation strategies, all the beta-adrenergic antagonists studied could be resolved into their enantiomers, although the three methods were carried out without extensive optimization. The protein and molecularly imprinted phases gave the highest selectivities whereas employing cyclodextrins resulted in the highest separation efficiency. Proteins and cyclodextrins are primarily natural products, albeit the cyclodextrins can be derivatized. In contrast, the molecularly imprinted chiral stationary phase can be highly customized when produced. PMID- 9221875 TI - Electrokinetic chromatography employing an anionic and a cationic beta cyclodextrin derivative. AB - beta-Cyclodextrin, 1, can act as a chiral buffer additive in capillary zone electrophoresis (CZE) owing to its ability to form diastereomeric complexes with the enantiomers of ionic compounds. Reaction of 1 with 2,3-epoxy propyltrimethylammonium chloride gave preponderantly 6-O-(2-hydroxy-3 trimethylammoniopropyl) derivatives (2) of varying degrees of substitution (d.s.). Analogous reaction of 1 with 1,3-propanesultone yielded 6-O-(sulfo-n propyl) derivatives (SPE-beta-CD, 3) of varying d.s. The purity and the d.s. of 2 and 3 were determined by 1H NMR and electrospray ionization mass spectrometry (ESI-MS). These charged beta-cyclodextrin derivatives were used for the chromatographic enantiomer separation of a series of neutral barbiturates, of chlorthalidone, terbutaline, warfarin, salbutamol and brompheniramine by cyclodextrin electrokinetic chromatography (CD-EKC). During the separation of chlorthalidone with 3 as chiral additive, a reversible interconversion of the enantiomers (enantiomerization) was observed at temperatures below 20 degrees C. PMID- 9221876 TI - Histamine-modified beta-cyclodextrins for the enantiomeric separation of dansyl amino acids in capillary electrophoresis. AB - Two novel monosubstituted beta-cyclodextrins (CD) bearing the histamine moiety linked to the upper CD rim, either through the amino group or the imidazole nitrogen 1N, CD-hm and CD-mh, were successfully used as chiral selectors in capillary electrophoresis for the enantiomeric discrimination of dansyl (Dns) amino acids. Good results were obtained by using low concentrations of the selectors (1-3 mM). The effect of pH on the chiral discrimination was studied in order to modulate the number and the position of the positive charges. By increasing the pH from 5 to 7.5, chiral discrimination decreased along with the deprotonation of the imidazolyl moiety. Inversion of the migration order was observed with the two CDs, depending on the relative position of the charged moieties on the upper rim. Ion pair interaction coupled to inclusion complexation seems to account for the discrimination process. The effects of the temperature, CD concentration and capillary length on chiral resolution were also examined. PMID- 9221877 TI - Comparison of alkylglycoside surfactants in enantioseparation by capillary electrophoresis. AB - Three alkylglycoside surfactants, namely n-octyl-beta-D-glucopyranoside (OG), n nonyl-beta-D-glucopyranoside (NG), and n-octyl-beta-D-maltopyranoside (OM), were compared in the enantiomeric separation of dansyl amino acids, binaphthyl phosphate, bupivacaine and warfarin. While only OM exhibited an enantioselectivity toward warfarin, bupivacaine, and dansyl tryptophan, all three surfactants were effective in the enantiomeric resolution of napthyl phosphate and other dansyl amino acids. With the exception of naphthyl phosphate, which could be resolved enantiomerically with OM at surfactant concentrations below the CMC, all solutes required surfactant concentrations greater than the CMC value. This was attributed to the strong hydrophobic association of napthyl phosphate with the OM monomers and to the presence of maltoside residue in the OM surfactant. In general, the optimum surfactant concentration needed for maximum enantiomeric resolution was an inverse function of the hydrophobic character of the solute. Under a given set of conditions, the enantiomeric resolution exhibited by the alkylglycoside surfactants was largely influenced by the extent and loci of solute solubilization into the micelle, and by the nature of the chiral sugar head group of the surfactant. PMID- 9221874 TI - Use of a zwitterionic cyclodextrin as a chiral agent for the separation of enantiomers by capillary electrophoresis. AB - The purity and enantioselectivity of a novel chiral agent, the zwitterionic mono (6-delta-glutamylamino-6-deoxy)-beta-cyclodextrin (beta-CD-Glu), were studied by capillary electrophoresis. Chiral separation of the enantiomers of chlorthalidone was obtained at pH 2.3, a pH at which beta-CD-Glu is partially protonated. Comparison with the cationic mono-(6-amino-6-deoxy)-beta-cyclodextrin (beta-CD NH2) enantioselectivity clearly shows that the greater the difference in mobility between the free analyte and the analyte-cyclodextrin complex, the better the resolution. Hydrobenzoin enantiomers were separated at pH 11.2, a pH at which beta-CD-Glu is anionic. Under these conditions, the migration order was opposite to that observed in the presence of beta-CD-NH2 at pH 2.3. When no separation was obtained directly with beta-CD-Glu, a dual cyclodextrin system was developed. Carprofen enantiomers were resolved at pH 2.3 in the presence of a beta-CD Glu/trimethyl-beta-cyclodextrin (TM-beta-CD) system in which the charged CD confers a non-zero mobility to the analyte, while the neutral CD allows chiral recognition. PMID- 9221878 TI - Determination of association constants of dansyl-amino acids and beta cyclodextrin in N-methylformamide by capillary electrophoresis. AB - The use of nonaqueous background electrolytes in capillary electrophoresis (CE) is a promising new trend which should widen the scope of this technique. We demonstrate the chiral separation of dansyl-amino acids (Dns-AAs) in N methylformamide (NMF) using beta-cyclodextrin (beta-CD) as chiral selector. The solubility of beta-CD is much better in NMF than in water, allowing high concentration of the chiral selector and successful enantioseparation despite the weak host-guest interaction between the Dns-AAs and beta-CD. The association constants for the complexation between Dns-AAs and beta-CD could be calculated from the electrophoretic mobilities, with attention paid to the change in viscosity of the electrolyte upon addition of beta-CD. The association constants ranged between 2 and 13 M(-1). PMID- 9221879 TI - Separation of enantiomers of drugs by capillary electrophoresis. Part 4: hydroxypropyl-gamma-cyclodextrin as chiral solvating agent. AB - In an extended chiral drug screening program, enantioseparation of 86 racemic drugs was tested with hydroxypropyl-gamma-cyclodextrin as chiral solvating agent (CSA). A total of 30 drugs out of 86 could be resolved in this straightforward approach. The number of experiments performed under identical conditions allows a statistical treatment of the data. The enantioseparation of the analytes is correlated with their interaction strength with the CSA. Hence, the concentration of the CSA is a crucial parameter for optimization of the enantioseparation, as shown by a subset of 23 examples. PMID- 9221880 TI - Central composite design in the chiral analysis of amphetamines by capillary electrophoresis. AB - We have studied a central composite design for the chiral separation of amphetamines using capillary electrophoresis. Five variables, i.e., buffer concentration, pH, chiral selector concentration, temperature, and applied voltage, were investigated. Enantiomeric resolutions as well as analysis time and generated power were established as responses for each experiment. A model of each response was obtained by multiple regression of a quadratic-degree mathematical expression. From the models, we determined the most favorable conditions for the chiral separation by optimizing the resolution and setting the other responses at threshold values. Results were compared with a previous study in which a systematic investigation of the operating parameters was carried out. In order to visualize the robustness of the method, response surfaces were drawn for the significant variables. We have concluded that experimental designs offer a rapid means of optimizing several variables and provide an efficient test for the robustness of the analytical method. PMID- 9221881 TI - Chiral interactions in capillary zone electrophoresis: computer simulation and comparison with experiment. AB - Chiral interaction in capillary electrophoresis can be modeled using pK values, mobilities of analytes, and their formation constants with the chiral selector. An existing steady-state simulation program for CE (HPCESIM) was recently extended with a chiral submenu involving the chiral parameters listed above. These were experimentally determined in both our laboratories for mandelic acid and terbutaline using hydroxypropylated beta-cyclodextrin as chiral selector. A comparison was made between both sets of parameters and between experimental electropherograms and those obtained from simulation. Error analysis of the results indicate the sensitivity of the obtained results. PMID- 9221882 TI - Chiral separation of DL-peptides and enantioselective interactions between teicoplanin and D-peptides in capillary electrophoresis. AB - Teicoplanin has been evaluated as a selector for enantioseparation of di- and tripeptide derivatives in capillary electrophoresis. Separation variables such as type of buffer, pH, concentrations of teicoplanin and organic modifier were examined. Optimal separation conditions were obtained by means of factorial design experiments. The effects of teicoplanin concentrations below and above its critical micellar concentration (CMC) and of acetonitrile (ACN) on the separation were demonstrated. The use of a high concentration of ACN resulted not only in increased selectivity, but also in improved separation efficiency. Electroosmotic flow was observed to be largely independent of the concentrations of teicoplanin under the optimized conditions. Good repeatability of migration times was obtained. The interactions between teicoplanin and D and L peptides were studied, and it was found that, for some peptides, teicoplanin exhibited enantioselective interaction only with the D-form. Somewhat lower separation efficiencies were thus observed for the strongest interacting (D-form) peptides. Chiral separation of 15 DL-peptide derivatives was achieved in less than 10 min. PMID- 9221883 TI - Enantioresolution of disopyramide by capillary affinity electrokinetic chromatography with human alpha1-acid glycoprotein (AGP) as chiral selector applying a partial filling technique. AB - A method using alpha1-acid glycoprotein (AGP) as chiral selector for disopyramide by means of affinity electrokinetic chromatography has been developed. In order to avoid UV absorbance interferences, less than the effective length of the capillary was filled with the chiral selector. The electrophoretic conditions were chosen to give opposite migration directions for the chiral selector and the analyte; AGP migrated away from the detector. Enantiomers of disopyramide were separated on a methylcellulose-coated capillary with 20 cm length to the detector. The enantioresolution of the solute was affected by the concentration of the chiral selector, the plug length of the selector in the capillary, and the applied voltage. Resolution factors and migration times decreased with reduction of the plug length, while the efficiency of the separation system and peak performance were improved by decreasing the separation zone. A special feature of the technique is an enhanced selectivity due to increasing separation of the enantiomers when the fastest has migrated from the selector zone, while the second one still is retained. Equations relating selectivity and resolution with the difference in effective plug lengths between the two enantiomers are developed. Optimized conditions yielding complete resolution, requiring an 0.75 mM AGP plug of only 4.5 cm effective length, also gave high efficiencies (about 400,000 plates/m) for both enantiomer peaks. PMID- 9221884 TI - A quantitative relationship between capacity factor and selector concentration in capillary electrophoresis and high performance liquid chromatography: evidence from the enantioselective resolution of benzoin using human serum albumin as a chiral selector. AB - Many selectors are used both in pressure-driven liquid chromatography (LC) and in electrokinetic chromatography (EKC), particularly chiral species such as cyclodextrins and proteins. It should be possible to readily apply information gleaned using one technique to the other, since in both techniques the underlying molecular interactions which lead to separations are expected to be the same. Superficially this may be the case, but an exact transfer of operating conditions, i.e., background electrolyte (BGE) composition/mobile phase composition, assuming that these meet certain minimum requirements for each technique, is not often possible. To investigate the reason for this we have measured retention (k') of a neutral solute (racemic benzoin) in HPLC and EKC using an identical range of BGE/mobile phase conditions in both techniques. The selector used was human serum albumin. The k' measurements obtained for each benzoin enantiomer were consistently higher in HPLC than in EKC. This can be explained very simply if one considers that retention in both systems is related to the selector concentration [S], by the expression k'=K[S], where K is the affinity constant. In EKC, [S] is simply the concentration of free selector in the BGE, while in LC, [S] = m(p)/Vo, where m(p) is the number of moles of accessible selector, and Vo is the column void volume. In LC, [S] is generally considerably higher than in EKC, leading to larger values of k'. PMID- 9221885 TI - Application of capillary electrophoresis for the analysis of chiral drugs in biological fluids. PMID- 9221886 TI - Chiral o-phthaldialdehyde reagents for fluorogenic on-column labeling of D- and L amino acids in micellar electrokinetic chromatography. AB - Following a recent communication from this laboratory (A. Tivesten et al., J. High Resol. Chromatogr. 1996, 19, 229-233) where on-column chiral derivatization of D- and L-amino acids in micellar electrokinetic chromatography (MEKC) was demonstrated for the first time, we now present further details of the labeling procedure. The basis of the method is the consecutive injection of a sample and the reagent onto the capillary as two discrete plugs. By utilizing their difference in mobility, the zones are mixed by the electrophoretic process in a controllable way. In this way the amino acids are both derivatized within a few seconds and subsequently separated in a single step. Compared with pre-column derivatization, dilution of the original sample is minimized, which is why the method is highly useful for microchemical analytical work, i.e., labeling of nano to picoliter samples. Four different chiral thiols were compared in this study, 2,3,4,6-tetra-O-acetyl-1-thio-beta-D-glucopyranose (TATG), N-acetyl-L-cysteine (AC), N-acetyl-D-penicillamine (AP), and N-isobutyryl-L-cysteine (IBC). Together with o-phthaldialdehyde (OPA) these constitute the chiral reagent. The reaction rate as well as the spectroscopic and chromatographic properties of the formed derivatives were examined. It was found that the fastest reaction is obtained with OPA/TATG, as was the case with L-alanine (L-ala), and that the rate is greatly affected by the presence and concentration of acetonitrile or methanol. Moreover, OPA/TATG yields superior resolution of D- and L-amino acids over the other OPA/thiol combinations in a sodium dodecyl sulfate (SDS) micellar buffer, whereas the OPA/AC and OPA/IBC-amino acid derivatives have a higher fluorescence quantum yield. With laser-induced fluorescence detection (He-Cd, 325 nm) the mass limit of detection is at the low amol level. PMID- 9221887 TI - Separation of synthesized enantiomers and diastereomers of thiino-[4,3-b]pyrrole 2,3-dione derivatives and 2H-thiopyrans by capillary electrophoresis. AB - Capillary electrophoresis was used to separate diastereomers and enantiomers of synthesized thiopyrans and thiinopyrroles. Separation conditions were optimized by varying the buffer parameters, i.e., kind and concentration of chiral selector, pH, main buffering component, micelle forming additives, and content of organic solvents. The interaction of thiopyrans and thiinopyrroles with beta- and gamma-cyclodextrins results in the separation of enantiomers and diastereomers. The magnitude of the resolution depends on the spatial requirements of the different substituents of the analytes and the dimensions of the cyclodextrin cavity. For well-separated enantiomers or diastereomers the identification and characterization of degradation products was possible. PMID- 9221889 TI - Determination of the enantiomeric purity of N-propionyl-6,7-dimethoxy-2 aminotetralin by cyclodextrin-modified micellar electrokinetic chromatography. AB - N-Propionyl-6,7-dimethoxy-2-aminotetralin (Z12231A), a useful intermediate in the synthesis of dopaminergic agonists, was resolved into its enantiomers by high performance capillary electrophoresis. Cyclodextrin-modified micellar electrokinetic chromatography was employed with a combination of chiral selectors both in the aqueous phase (a hydroxyalkyl-beta-cyclodextrin) and in the micellar phase (sodium taurodeoxycholate). The absolute amount of the two chiral selectors as well as their ratio were found to be the most critical parameters in order to optimize separation. A resolution factor >3.5 was obtained, allowing a high amount of solute to be loaded in order to improve the detection limit. The reproducibility of the assay was also evaluated. PMID- 9221888 TI - Resolution of acylated dipeptide stereoisomers by capillary electrophoresis using sulfobutylether derivatized beta-cyclodextrin. AB - The separation of enantiomerically and diastereomerically related stereoisomers of acylated Asp-Phe dipeptides was explored using capillary electrophoresis (CE). This series of dipeptides included the alpha-L,L parent compound and the three other potential Asp containing stereoisomers (alpha-D,D, alpha-L,D, and alpha D,L), as well the four possible isoAsp containing stereoisomers (beta-L,L, beta D,D, beta-L,D and beta-D,L). The separation of these substances was explored using both neutral and charged cyclodextrins as the stereoisomer selector added to the running electrolyte. The major experimental parameters investigated included pH, the cyclodextrin type, and the cyclodextrin concentration. Due to differences in the pKa values of the carboxylic acid groups, adjustment of the separation buffer to between pH 3.0 and 4.0 provided for sufficient electrophoretic mobility differences to result in excellent separations of the diastereomerically related peptides in this pH region. The resolution of the enantiomerically related peptide stereoisomers was accomplished using low concentrations (1 mM) of the anionic cyclodextrin derivative, sulfobutylether beta-cyclodextrin (SBE-beta-CD). This negatively charged cyclodextrin was found to be superior for the resolution of the enantiomerically related peptides as compared to native beta-cyclodextrin or the neutral derivatives, dimethyl beta cyclodextrin and hydroxypropyl beta-cyclodextrin. An alternative approach using anionic or neutral surfactants in conjunction with the SBE-beta-CDs was also explored and found to be successful but problematic. PMID- 9221890 TI - Enantiomer separation of disopyramide with capillary electrophoresis using various cyclodextrins. AB - Enantiomers of disopyramide display different biological actions, and therefore chiral selective analysis is necessary. Fifteen different cyclodextrins (CDs) and CD derivatives were tested as capillary electrophoresis (CE) additives for the chiral separation of disopyramide. Eleven types of CDs showed chiral recognition features and four types had a baseline or close to baseline separation. The best resolution (Rs = 3.0) was with 15 mM carboxymethylated beta-CD (pH 4.9). A sharp decrease in the selectivity of gamma-phosphate (gamma-PhoCD) was observed in the pH range of 2-3, indicating a structural change of gamma-PhoCD. The enantiomers of disopyramide were separated in its ionized as well as neutral forms using acidic substituted CDs. The results show that the size of the CD cavity can not be used as a guide to estimate chiral separations, suggesting a more complex separation mechanism of these CDs towards disopyramide. PMID- 9221891 TI - Enantiomer separation of denopamine by capillary electrophoresis with charged and uncharged cyclodextrins. AB - Direct separation of enantiomers of denopamine was investigated by capillary electrophoresis employing charged and uncharged cyclodextrin (CD) derivatives. Uncharged beta-type CDs, having hydrophobic groups, were essential for the enantioseparation of denopamine; of these, especially dimethyl-beta-CD was effective. Among charged CDs, gamma-type as well as beta-type CDs were found effective for the enantioseparation of denopamine. Reversal of migration order of R-form (active) and S-form enantiomers was investigated by using two types of coated capillaries: (i) an amine capillary with an inner wall coated with dimethylamino groups, and (ii) a polyacrylamide-coated capillary. Manipulation of migration order could be easily performed by selecting suitable capillaries, buffer pH, and CDs. PMID- 9221892 TI - Enantioseparation of nonsteroidal anti-inflammatory drugs by capillary electrophoresis using mixtures of anionic and uncharged beta-cyclodextrins as chiral additives. AB - Nine nonsteroidal anti-inflammatory drugs (NSAIDs) were enantioseparated by capillary electrophoresis using an anionic cyclodextrin derivative (sulfobutyl ether beta-cyclodextrin or carboxymethyl-beta-cyclodextrin) in combination with a neutral cyclodextrin as chiral additives to a pH 3 phosphoric acid triethanolamine buffer. In the presence of a negatively charged cyclodextrin, the analytes were given an appropriate mobility but relatively low enantioselectivities were generally obtained when such a cyclodextrin was the only selector added to the buffer. The addition of an uncharged cyclodextrin, such as the native beta-cyclodextrin or one of its derivatives (dimethyl-, trimethyl- and hydroxypropyl-beta-cyclodextrin), to this kind of buffer containing an anionic cyclodextrin, was found to give rise to considerable improvement in chiral resolution for all compounds studied. Resolution and analysis time were optimized by varying the nature and concentration of the two cyclodextrins. The best compromise was usually achieved by the simultaneous addition of sulfobutyl ether beta-cyclodextrin and trimethyl-beta-cyclodextrin. Under optimum conditions, the enantiomers of all NSAIDs examined could be completely separated (most often with resolution values higher than 5) in short analysis times (generally lower than 15 min). PMID- 9221893 TI - Enantioseparation of local anaesthetic drugs by capillary zone electrophoresis with cyclodextrins as chiral selectors using a partial filling technique. AB - The enantiomers of prilocaine were successfully resolved with alpha-cyclodextrin, and those of mepivacaine and bupivacaine, with methyl-beta-cyclodextrin as chiral selectors, by means of capillary zone electrophoresis (CZE) employing a partial filling technique. By this separation mode, a discontinous separation zone is formed in the capillary. Prior to application of the actual drug substance, the capillary is partially filled with the separation solution. During the enantioseparation both ends of the capillary are dipped into the running buffer solution, i.e., without chiral selector. The consumption of chiral selector is thus very low, less than a microliter per run. The repeatibility of the electrophoretic mobility of the enantiomers was better than 1.2% relative standard deviation (RSD). The effect of the length of the separation zone on the resolution of the enantiomers was studied. The application time of the chiral selector, instead of the selector concentration, was varied in order to improve and regulate the enantioresolution and reduce consumption of the chiral selector as much as possible. It was found that the enantioseparations were directly affected by the length of the separation zone, and there was a minimal plug length where complete enantioresolution was achieved. PMID- 9221894 TI - Enantioselective determination of methadone and its main metabolite 2-ethylidene 1,5-dimethyl-3,3-diphenylpyrrolidine (EDDP) in serum, urine and hair by capillary electrophoresis. AB - A capillary electrophoresis method has been developed for the enantioselective determination of methadone and its main metabolite 2-ethylidene-1,5-dimethyl-3,3 diphenylpyrrolidine (EDDP) in serum and urine. After addition of the internal standard diphenhydramine, the serum and urine samples were extracted at pH 9-10 with n-hexane. Different cyclodextrins were tested for use as chiral selectors. The best results for simultaneous separation of methadone and EDDP were obtained using heptakis-(2,6-di-O-methyl)-beta-cyclodextrin (DIMEB). Data on the accuracy and precision of the method are presented. The method was applied to serum, urine, and hair samples, from individuals undergoing methadone therapy, under consideration of multi-drug abuse. PMID- 9221895 TI - Enantioselective determination of 3,4-methylene-dioxymethamphetamine and two of its metabolites in human urine by cyclodextrin-modified capillary zone electrophoresis. AB - Using capillary zone electrophoresis with a phosphate buffer at pH 2.5 containing 30 mM (2-hydroxypropyl)-beta-cyclodextrin as chiral selector, the simultaneous separation of the enantiomers of 3,4-methylenedioxymethamphetamine (MDMA or Ecstasy) and its two metabolites 4-hydroxy-3-methoxymethamphetamine (HMMA) and 3,4-methylenedioxyamphetamine (MDA) in human urine is reported. The assay described is based upon enzymatic hydrolysis of conjugated HMMA (major urinary metabolite) and solid-phase extraction followed by injection of a few nL of the extract onto a 50 microm internal diameter (ID) fused-silica capillary of 60 cm length. Solutes are detected via on-column absorbance at 195 nm. For 375 ng/mL drug levels, intraday and interday imprecision is < 4%. With 5 mL urine samples, the detection limit is in the 20-50 ng/mL range. Via analysis of the urines of two patients, the metabolism of MDMA is demonstrated to be enantioselective, with significantly higher urinary amounts of R-(-)-MDMA being excreted compared to S (+)-MDMA. Within 72h after drug administration one patient was determined to excrete 42.28 and 10.16% of the racemic MDMA dose (1.5 mg/kg body weight) as R-( ) and S-(+)-MDMA enantiomers, respectively. Corresponding values for the second subject were found to be 28.63 and 9.34%. The metabolism of the enantiomers of the two metabolites showed interindividual differences. The first and second detected HMMA enantiomers represented 3.79 and 5.42% (first subject) and 8.51 and 4.36% (second), respectively, of the administered MDMA dose. For the MDA enantiomers, corresponding values were 2.44, 1.76, 0.75, and 0.79%, respectively. PMID- 9221896 TI - Identification of preferentially expressed cochlear genes by systematic sequencing of a rat cochlea cDNA library. AB - 107 expressed sequence tags (ESTs) from a rat cochlea cDNA library were identified by systematic sequencing coupled to database selection and RT-PCR analysis of novel sequences. This approach led us to select a clone, pCO8, showing no significant homology with any database sequence, that corresponds to a mRNA whose expression is restricted to the cochlea, except for traces detected in brain. Additional clones with novel sequences enriched in the cochlea were also found. ESTs bearing significant homologies with database sequences (63 out of 107) were classified according to the putatively encoded protein. They include tissue-specific genes not previously described in the cochlea as well as known genes from other species. We performed in situ hybridization in cochlear tissues to localize the pCO8 mRNA and that of clone pCO6 which is 100% homologous to the delayed rectifier potassium channel drk1. We found that both mRNAs were exclusively expressed in the cellular body of the primary auditory neurons from the spiral ganglion of the cochlea. These results indicate that this approach is an efficient way to identify novel genes that could be of importance in cochlear function. PMID- 9221898 TI - Adrenergic differentiation potential in PC12 cells: influence of sodium butyrate and dexamethasone. AB - The ability of sodium butyrate and dexamethasone to promote adrenergic differentiation in PC12 cells was examined using the gene encoding the epinephrine biosynthetic enzyme, phenylethanolamine N-methyltransferase (PNMT), as a marker. Sodium butyrate and dexamethasone independently stimulated expression of PNMT mRNA in PC12 cells, and the combined action of these drugs led to synergistic activation of the PNMT gene. Despite the induction of the PNMT gene, epinephrine is not produced in these cells, in part due to the absence of a corresponding induction in PNMT enzymatic activity. Another contributing factor appears to be a reduction in the precursor catecholamines, norepinephrine and dopamine, in the presence of sodium butyrate. Thus, while sodium butyrate and dexamethasone can induce PNMT gene expression, treatment of PC12 cells with these drugs appears insufficient for full acquisition of the adrenergic phenotype. PMID- 9221897 TI - The rat galanin-gene promoter: response to members of the nuclear hormone receptor family, phorbol ester and forskolin. AB - We have cloned a rat genomic DNA fragment of approximately 12.5 kb. Nine kb of the cloned fragment lie in the 5'-flanking region of the gene and contain the promoter elements, while the remaining 3.5 kb contain the first four complete exons, the first three introns, and part of the fourth intron of the rat galanin gene. We have partially analysed some of the elements within the proximal sequence of this promoter which may influence the transcriptional regulation of the rat galanin gene. The rat galanin-gene promoter contains many regions which share homology with both the human and the bovine galanin genes and certain cis elements appear to be conserved among the three species. In an attempt to test whether some of these elements are functional in the rat gene, transient transfection studies were carried out in selected cell lines. Estrogen, thyroid hormone and retinoic acid all showed a minimal degree of promoter stimulation when the rat galanin-gene promoter was co-transfected with the appropriate hormone receptors in Neuro 2A cells, while co-transfection of the nuclear orphan receptor ELP1 was able to stimulate transcription of a galanin promoter-driven reporter-gene construct (-374 bp) by 35-fold. The galanin promoter mediated a 3-4 fold induction in response to forskolin or TPA. Deletion of a 5-bp element at -50 bp from the start of transcription was able to greatly reduce the forskolin response but not the TPA response. These results point to several elements that may be targets of transcription factors linked to extracellular stimuli. PMID- 9221899 TI - Changes in expression of delta FosB and the Fos family proteins following NMDA receptor activation in the rat striatum. AB - Receptor-induced expression of transcription factors of the activator protein-1 (AP-1) family in neurons occurs in a unique temporal pattern which regulates subsequent downstream gene expression. We investigated the expression of the Fos family proteins following injection of the NMDA receptor agonist quinolinic acid (QA) into the rat striatum. The c-Fos protein is rapidly and transiently expressed, followed by the sequential and overlapping expression in the same striatal neurons of FosB, from 4 to 8 h post-lesion and delta FosB from 6 h to beyond 30 h post-lesion. Analysis confirms that mRNA transcripts of both fosB and alternatively spliced delta fosB are expressed in the striatum after QA lesion. The Fos-related antigens Fra-1 and Fra-2 and three previously uncharacterized c Fos-related proteins were additionally found in the striatum which do not increase following lesion. These proteins are related to the highly conserved DNA binding domain of c-Fos but are not immunologically related to the FosB protein as has been previously reported for proteins induced following chronic stimulation of the striatum. We additionally demonstrate that the c-Fos and delta FosB proteins expressed following QA lesion bind to the functional AP-1 site in the promoter of the nerve growth factor (NGF) gene, the regulation of which temporally and spatially coincides with the AP-1 protein increases in the QA lesioned striatum. However, the levels of binding to the NGF AP-1 site do not increase throughout time following lesion despite the induced expression of Fos family proteins, suggesting that the regulation of the NGF gene in this paradigm does not simply involve increased binding to the AP-1 site in the NGF gene promoter. PMID- 9221900 TI - Activation of protein kinase A prevents the ethanol-induced up-regulation of delta-opioid receptor mRNA in NG108-15 cells. AB - We have used a sensitive solution hybridization assay with a riboprobe transcribed from the coding sequence of the delta-opioid receptor gene (DOR) to study the up-regulation of the DOR mRNA by ethanol in NG108-15 cells. Exposure of the cells to compounds that increase cAMP levels (forskolin, forskolin + IBMX, or dibutyryl cAMP) resulted in the attenuation of ethanol-induced up-regulation of DOR mRNA. The inactive analogue of forskolin, 1,9-dideoxy forskolin had no effect. Northern blot analysis of RNA extracts from ethanol-, forskolin- or ethanol + forskolin-treated cells showed proportional changes in each of the multiple DOR mRNA bands, so that no difference was observed in the fraction of the total hybridization signal produced by each band of the DOR mRNA. In the absence of ethanol, forskolin or dibutyryl cAMP reduced the basal levels of DOR mRNA. The cAMP analogue (Rp)-cAMPS, a protein kinase A (PKA) inhibitor, increased DOR mRNA levels. However, the combination of (Rp)-cAMPS and ethanol did not further increase DOR mRNA levels compared to ethanol or (Rp)-cAMPS alone. Signaling through cAMP and PKA down-regulates DOR mRNA levels. The ethanol induced increase in DOR mRNA levels in NG108-15 cells appears to be mediated via a reduction of PKA. PMID- 9221901 TI - Molecular mechanism of regulation of pentylenetetrazol-induced calcium entry by 3'-untranslated region of a seizure-related cDNA, PTZ-17, in Xenopus oocytes. AB - To determine the molecular mechanism of regulation of pentylenetetrazol (PTZ) induced calcium entry by the seizure-related gene, PTZ-17, the role of the 3' untranslated region (3'UTR) and also interaction between 3'UTR and intracellular factors were investigated. PTZ-induced calcium inward current in Xenopus oocytes injected with PTZ-17 RNA varied in magnitude among strains of mice: RNA derived from the DBA/2 mouse, which has a high susceptibility to convulsions, showed the largest current and that from the BALB/c mouse with a low susceptibility to convulsions showed no PTZ response. The sequence of 3'UTR showed alterations among mouse strains: 3'UTR of BALB/c showed a sequence alteration from T to G and that of DBA/2 showed a GTG insertion compared with that of B6. The 3'UTR also regulated the translation of chloramphenicol acetyltransferase (CAT) RNA depending on its sequence. A particular region within the 3'UTR demonstrated interaction with 60- and 47-kDa proteins. Sequence alterations in this region corresponded to disappearance or increase in PTZ-induced calcium entry. These findings suggest that a particular region within 3'UTR of the seizure-related gene, PTZ-17, is involved in PTZ-induced calcium entry via interaction between mRNA and specific RNA-binding proteins. PMID- 9221902 TI - Molecular characterization and pharmacological properties of the human P2X3 purinoceptor. AB - Using PCR and library screening techniques, a cDNA encoding an ATP ligand-gated channel has been isolated from human heart. The full-length cDNA encodes a protein 397 amino acids long which shows a high amino-acid sequence identity with the rat P2X3 purinoceptor (93%). By fluorescence in situ hybridization, the human P2X3 gene has been mapped to region q12 of chromosome 11. Tissue distribution analysis of human P2X3 receptor mRNA shows a restricted expression pattern, i.e. transcripts are limited to the spinal cord and heart. This result contrasts with the distribution of the rat P2X3 receptor which was detected exclusively in sensory neurons of trigeminal, dorsal root and nodose ganglia. Heterologous expression of human P2X3 cRNA in Xenopus oocytes generates a fast desensitizing ATP-activated channel with pharmacological properties resembling the profile of the rat homologue receptor. Thus, the order of agonist potency is 2MeSATP > ATP > alphabeta-meATP > CTP > betagamma-meATP approximately ADP. Moreover, ATP-evoked currents on human P2X3 receptor are efficiently blocked in a reversible manner by the purinoceptor antagonists, suramin and PPADS. PMID- 9221903 TI - Cloning and developmental expression of 5-HT1A receptor gene in Xenopus laevis. AB - The aim of our work is to investigate the potential involvement of serotonin and its G-protein-coupled receptors in neural differentiation or other developmental processes in Xenopus laevis. By using a RT-PCR strategy, we isolated a cDNA fragment from X. laevis brain showing high amino-acid similarity with the mammalian 5-HT1A receptor. We used this fragment to isolate a cDNA clone containing a single ORF of 408 amino-acids with an overall amino-acid identity of 73% with the human and rat 5-HT1A receptor. This structural similarity suggests that this clone encodes the Xenopus homolog of the mammalian 5-HT1A receptor (X5 HT1A). In order to establish a possible role for this receptor in development, we analyzed the pattern of its gene expression during embryogenesis, larval stages and in adult brain by in situ hybridization. The first signal of mRNA expression appears in the rostral part of brain stem at stage 22, when the first neurons start differentiation [38,21]. In later stages of development, the cells expressing X5-HT1A transcripts appear to correspond to serotonergic neurons. By stage 41, X5-HT1A mRNA is also detected in the inner nuclear layer (INL) of the developing retina. This pattern of expression is maintained until stage 46, i.e. at the beginning of metamorphosis. In adult, additional brain areas express X5 HT1A mRNA, particularly in telencephalon, diencephalon and mesencephalon. On the whole, our data show that the X5-HT1A receptor mRNA is developmentally regulated, with expression first appearing in differentiating serotonergic neurons, where this receptor may mediate, through an autocrine regulatory pathway, the trophic action of serotonin on developing serotonergic system. PMID- 9221904 TI - Regulation of preprotachykinin-A mRNA in genetic hypertensive and normotensive rats. AB - It is well-known that central administration of tachykinins (Tks) inhibit salt intake in rats. Recent studies have shown that conditions that arouse salt appetite, such as adrenalectomy and sodium depletion, induce a decrease in preprotachykinin-A (PPT-A) mRNA in discrete regions of the rat brain, suggesting that reduced levels of PPT-A mRNA in the brain may have a permissive role on the expression of salt appetite. It has also been shown that spontaneously hypertensive rats (SHR) show higher avidity for salty solutions than their normotensive control Wistar-Kyoto (WKY) rats. In this regard, the present study tested whether SHR and WKY rats differ in expression of the gene coding for PPT A, the precursor for Tks peptides. Using semi-quantitative in situ hybridization histochemistry, we examined the level of PPT-A mRNA in discrete rat brain regions of SHR and WKY rats under no treatment, after 1 or 3 days of Na+ depletion. Levels of PPT-A mRNA were analysed in the olfactory tubercle (Tu), in the lateral olfactory tubercle (LOT), in the dorsal and ventral caudate putamen (d/v CPu), in the medial preoptic area (mPOA), in the bed nucleus of the stria terminalis (BNST), in the habenula (Hb) and in the postero-dorsal part of the amygdala (MePD). Semi-quantitative analysis of silver grains revealed a 27.5% lower expression of the PPT-A mRNA levels in SHR opposite to WKY rats under no treatment in v-CPu, mPOA, BNST and Hb. 1 day of Na+ depletion reduced PPT-A mRNA levels when opposite to Na+-repleted animals in Tu and mPOA in both SHR and WKY rats. On the other hand, when comparing SHR and WKY rats after 1 day of Na+ depletion, a 26% lower level of PPT-A mRNA was detected in Tu and d-CPu of SHR opposite to WKY rats whereas a 14% and an 18% lower level was detected in v-CPu and Hb, respectively. A lower expression of PPT-A mRNA in SHR compared to WKY rats was also found in BNST and MePD, although no statistical significance was detected in these two brain areas. In the last experiment, 3 days of Na+ depletion reduced PPT-A mRNA levels in mPOA while negligibly increased mRNA levels in d-CPu and v-CPu, in BNST, Hb and MePD, both in SHR and WKY rats. Conversely, when making comparisons between the two strains, a 35% lower level of PPT-A mRNA in SHR with respect to WKY rats was found after 3 days of Na+ depletion in d-CPu, v-CPu and mPOA. A lower gene expression, even though not statistically significant, was found in Tu, LOT, MePD. These findings show a consistent difference of PPT-A mRNA levels in discrete regions of the SHR brain opposite to WKY rats and confirm that 1 day of Na+ depletion reduces PPT-A mRNA in discrete brain regions. Since SHR are notoriously more salt-avid than WKY rats and Tks are potent inhibitors of sodium intake, the down-regulation of PPT-A mRNA may contribute to the higher natriophilia and, therefore, to the etiology of the hypertensive disease. PMID- 9221905 TI - Regulatory sequences for the transcription of the laminin B2 gene in astrocytes. AB - Astrocytes synthesize only the B2 chain of laminin and that this chain is sufficient to stimulate neurite outgrowth. In this study, we have examined laminin B1 and B2 promoter constructs in various cell types in order to understand the transcriptional regulation of laminin B2 gene in astrocytes. Comparison of nuclear factor binding by Southwestern analysis with the highly active B2 promoter fragment revealed different patterns of nuclear factor binding. In HepG2 cells, two proteins of 105 and 98 kDa were identified while, in primary astrocytes, human U251 and rat C6 glioma cells, a greater number of nuclear proteins ranging from 43 to 212 kDa were detected. The laminin B1 promoter construct was inactive in transient transfection experiments in astrocytes yet active in the HepG2 hepatoma cells which synthesize both the B1 and B2 chains. In contrast, the laminin B2 promoter construct was active in both astrocytes and HepG2 cells. These results are consistent with the lack of laminin B1 mRNA expression in astrocytes and suggest that the differential regulation of the laminin B1 and B2 gene is controlled at the transcriptional level. Delineation of the 5'-flanking regions responsible for basal levels of B2 laminin promoter activity revealed a silencer-like segment between -830 and -224 which reduced promoter activity. Deletion analysis further revealed that B2 laminin promoter possesses a highly active short promoter (-94 to +106) and basal transcriptional activity resides within -61 to +106. DNase 1 footprinting, gel shift competition assays and site-directed mutagenesis of a highly active short promoter revealed that this region contained binding sites for cell-type nuclear factors. The shortest construct containing only residues -21 to +106 was inactive in HepG2 and U251 glioma cells. In primary astrocytes, however, this construct showed a high level of transcriptional activity. Deletion of 47 bp (+59 to +106) in 5'-UTR completely blocked promoter activity in astrocytes confirming that this downstream region is important for transcriptional activity in primary astrocytes. Together, these results suggest that astrocytes may utilize mutually exclusive transcription factors and regulatory sequences, in addition to common factors in the control of the laminin B2 promoter. PMID- 9221906 TI - Acute effects of D1- and D2-receptor agonist and antagonist drugs on somatostatin binding, inhibition of adenylyl cyclase activity and accumulation of inositol 1,4,5-trisphosphate in the rat striatum. AB - A recent study carried out by our group demonstrated that exogenous dopamine increases the somatostatin (SS) receptor-effector system in the rat striatum. The present study examined the participation of the D1- and D2-dopaminergic systems in the modulation of the rat striatal SS receptor-effector system by use of the D1-receptor agonist and antagonist SKF 38393 and SCH 23390, respectively, and the D2-receptor agonist and antagonist bromocriptine and raclopride, respectively. In view of the rapid onset of dopamine action, the effect of dopaminergic agents on the SS mechanism of action were studied 3 h after their administration. SKF 38393 (4 mg/kg i.p.) or bromocriptine (2 mg/kg i.p.) administered to male Wistar rats increased the number of 125I-Tyr3-SMS receptors in the striatum (52 and 30%, respectively) without changing the affinity constant. The effect of SKF 38393 on 125I-Tyr3-SMS binding was antagonized by the D1-specific antagonist SCH 23390 (0.25 mg/kg i.p.) whereas the effect of bromocriptine was abolished by the D2 specific antagonist raclopride (5 mg/kg i.p.). No change in binding was produced when SKF 38393 or bromocriptine were added directly to the incubation medium. The acute systemic administration of SCH 23390 or raclopride alone had no effect on the binding of 125I-Tyr3-SMS to its receptors. The increase of the number of 125I Tyr3-SMS receptor induced by SKF 38393 or bromocriptine was accompanied by an increase in the capacity of SMS 201-995 to inhibit basal and forskolin (FK) stimulated adenylyl cyclase (AC) activity when compared to the control groups. In addition, the effect of SMS 201-995 on the mass accumulation of inositol 1,4,5 trisphosphate (IP3) was investigated. SKF 38393 as well as bromocriptine increased the capacity of SMS 201-995 to accumulate IP3 in the rat striatum although this effect was only statistically significant in the case of SKF 38393. These results suggest that the activation of D1 and D2 receptors increases the activity of the SS receptor-effector system, the effect being greater in the case of D1 receptors. These findings are consistent with a functional interaction between dopamine and SS in the rat striatum. PMID- 9221908 TI - Influence of fluoxetine and p-chloroamphetamine on the somatostatin receptor adenylyl cyclase system in the rat frontoparietal cortex. AB - There is evidence that suggests a reciprocal functional link between the serotonergic and the somatostatinergic system in the rat frontoparietal cortex. However, to date, the role of endogenous 5-hydroxytryptamine (serotonin) on the regulation of the somatostatin (SS) receptor-adenylyl cyclase (AC) system remains unclear. In the present study, the administration of fluoxetine (10 mg/kg i.p.), a 5-hydroxytryptamine uptake inhibitor in a single dose or administered daily for 14 days increased the number of specific [125I]Tyr11-SS receptors, with no change in the receptor affinity, in rat frontoparietal cortical membranes. However, the capacity of SS to inhibit forskolin (FK)-stimulated AC activity in these membranes was lower than in the control groups. The ability of the stable GTP analogue 5'-guanylylimidodiphosphate (Gpp(NH)p) to inhibit FK-stimulated AC activity in frontoparietal cortical membranes was also decreased in rats acutely and chronically treated with fluoxetine. p-Chloroamphetamine (5 mg/kg i.p.), which leads to a lasting reduction of 5-hydroxytryptamine innervation, administered on days 1, 3 and 5 and the rats sacrificed 1 or 3 weeks after the first injection, decreased the number of SS receptors without changing the receptor affinity. In this experimental group, SS also caused a significantly lower inhibition of FK-stimulated AC activity. p-Chloroamphetamine had no effect on the ability of Gpp(NH)p to inhibit FK-stimulated AC activity in frontoparietal cortical membranes at all the time periods studied. The present results suggest that under normal circumstances some SS receptors are under a tonic stimulatory control through the serotonergic system. PMID- 9221907 TI - Proteolytical processing of mutated human amyloid precursor protein in transgenic mice. AB - The evidence that betaA4 is central to the pathology of Alzheimer's disease (AD) came from the identification of several missense mutations in the amyloid precursor protein (APP) gene co-segregating with familial AD (FAD). In an attempt to study the proteolytical processing of mutated human APP in vivo, we have created transgenic mice expressing the human APP695 isoform with four FAD-linked mutations. Expression of the transgene was controlled by the promoter of the HMG CR gene. Human APP is expressed in the brain of transgenic mice as shown by Western blot and immunohistology. The proteolytic processing of human APP in the transgenic mice leads to the generation of C-terminal APP fragments as well as to the release of betaA4. Despite substantial amounts of betaA4 detected in the brain of the transgenic mice, neither signs of Alzheimer's disease-related pathology nor related behavioural deficits could be demonstrated. PMID- 9221909 TI - Regulation of preoptic area gonadotrophin-releasing hormone (GnRH) mRNA expression by gonadal steroids in the long-term gonadectomized male rat. AB - Testosterone exerts important feedback actions on the hypothalamus and pituitary of the male rat to control reproductive hormone secretion. Marked fluctuations occur in plasma-luteinizing hormone (LH) concentrations, hypothalamic gonadotrophin-releasing hormone (GnRH) content and GnRH mRNA expression following castration and it appears as though a stable post-castration equilibrium is not attained until 3-4 weeks after gonadectomy. In the present study, we have investigated the effects of long-term (7 week) gonadectomy on GnRH mRNA expression in the male rat and determined whether estrogen or androgen receptor mediated mechanisms are involved in regulating its expression. Accordingly, in situ hybridization was undertaken using a 35S-labelled antisense oligonucleotide probe complementary to bases 102-149 of the rat GnRH cDNA to quantify cellular GnRH mRNA expression in the medial septum (MS), diagonal band of Broca (DBB) and rostral preoptic area (rPOA) of intact males, rats gonadectomized for 7 weeks and gonadectomized animals implanted with silastic capsules containing testosterone (T), estrogen (E) or dihydrotestosterone (DHT). We found no difference between any of the treatment groups in the number of cells expressing GnRH mRNA in the MS/DBB or rPOA. Similarly, the GnRH mRNA content of cells in the MS/DBB was not different between the treatment groups. In contrast, cellular GnRH mRNA expression in the rPOA was elevated 7 weeks following castration (intact: 0.95 +/ 0.07 silver grains/microm2/cell; gonadectomized: 1.26 +/- 0.03; mean +/- S.E.M., P < 0.05) and this was restored to intact levels by either T (1.02 +/- 0.07) or E (1.02 +/- 0.08) treatment. DHT replacement had no effect on cellular levels of GnRH mRNA in gonadectomized rats (1.26 +/- 0.03). Frequency analysis of relative GnRH mRNA expression/cell showed that the rostral preoptic GnRH population responded to the steroid treatment in an homogeneous manner. These results show that GnRH mRNA expression is elevated specifically within the rPOA of the long term gonadectomized male rat when LH secretion has stabilized at a constant high level. Further, we show that the gonadal steroid regulation of cellular GnRH mRNA content at such time occurs only through an estrogen receptor-mediated pathway. PMID- 9221910 TI - Estrogen regulation of mu-opioid receptor mRNA in the forebrain of female rats. AB - Previous studies have suggested that opioids play a role in the regulation of reproductive behaviors in the female rat. The present study examined whether estrogen treatment alters mu-opioid receptor mRNA levels in different areas of the forebrain of ovariectomized (OVX) female rats using the in situ hybridization technique. We observed an increase in mu-opioid receptor mRNA levels in the ventromedial nucleus of the hypothalamus (VMH) and arcuate nucleus (ARN) after 48 h of 10 microg of 17-beta-estradiol-3-benzoate treatment when compared to OVX females. No effects of estrogen were observed on mu-opioid receptor mRNA levels in the posterior medial nucleus of the amygdala (MeAmyg), hippocampus, caudate putamen (CPu) or the medial habenula. Our result suggests that the estrogenic regulation of mu-opioid receptor in the CNS may in part be mediated by de novo synthesis and/or stability of the mu-opioid receptor message. PMID- 9221911 TI - Spatially restricted expression of fibroblast growth factor-10 mRNA in the rat brain. AB - Fibroblast growth factor (FGF)-10 is a novel member of the FGF family. Although FGF-10 mRNA was preferentially expressed in the lung, the mRNA was also expressed, although at low levels, in the brain. We examined the localization of FGF-10 mRNA along with FGF-7 mRNA in the rat brain by in situ hybridization. FGF 10 mRNA showed spatially restricted expression in some regions of the brain, including the hippocampus, thalamus, midbrain and brainstem, although FGF-7 mRNA was not expressed in any of the brain regions examined. FGF-10 mRNA was strongly expressed in several restricted nuclei, especially in motor nuclei, including the oculomotor nucleus, dorsal motor nucleus of vagus, motor trigeminal nucleus, facial nucleus and hypoglossal nucleus. This localization pattern was distinct from those of aFGF, bFGF FGF-5 and FGF-9 mRNAs reported previously. The cellular localization of FGF-10 mRNA showed that the mRNA in the brain was preferentially expressed in neurons but not in glial cells. The present findings indicate that FGF-10, an additional member of the FGF family expressed in the brain, has a distinct role in the brain. PMID- 9221912 TI - Cell-specific expression of beta-amyloid precursor protein isoform mRNAs and proteins in neurons and astrocytes. AB - The abnormal accumulation of beta-amyloid (A beta) in senile plaques appears to be a central pathological process in Alzheimer's disease. A beta is formed by proteolysis of beta-amyloid precursor protein (APP) with several isoforms generated by alternative splicing of exons 7, 8 and 15. A semi-quantitative reverse transcription (RT)-polymerase chain reaction (PCR) analysis showed that APP695 mRNA lacking exon 7 and 8 was most abundant in primary cultures of rat neurons, while APP770 and APP751 representing, respectively, the full length and exon 8 lacking isoforms predominated in cultured astroglial cells. Antisera AP-2 and AP-4 were produced by immunizing rabbits with keyhole limpet haemocyanin coupled with synthetic peptides representing KPI region APP301-316 and A beta region APP670-686 of APP770, respectively. These polyclonal antisera were purified against the corresponding peptide using affinity chromatography. Western blot analysis of homogenates of relatively enriched neuronal and astroglial cultures showed that these antibodies discretely stained bands of proteins in a cell-specific manner. Dot-blot analysis using AP-2, AP-4 and 22C11 antibodies indicated that, in comparison with neurons, cultured astrocytes contained 3-fold greater KPI-containing APP isoform proteins. The amount of total APP proteins, which include both KPI-containing and KPI-lacking APP isoforms, was approximately 90% higher in astrocytes than in neurons. Consistent with these in vitro findings in cultured astrocytes, in fimbria-fornix lesioned rat hippocampus, labelling with AP-2 antibody, which specifically reacts with KPI-containing APP proteins, was mainly observed in glial fibrillary acidic protein-positive reactive astrocytes in vivo. The results showed that APP isoforms are expressed in a cell type-specific manner in the brain and, since deposition of A beta is closely associated with the expression of KPI-containing APP isoforms, provide further evidence for the involvement of astrocytes in plaque biogenesis. PMID- 9221913 TI - mRNAs encoding urokinase-type plasminogen activator and plasminogen activator inhibitor-1 are elevated in the mouse brain following kainate-mediated excitation. AB - Urokinase-type plasminogen activator (uPA) is an inducible extracellular serine protease implicated in fibrinolysis and in tissue remodeling. Recently, we have localized uPA mRNA strictly in limbic structures and the parietal cortex of the adult mouse brain. Here, we tested whether the systemic treatment of mice with kainic acid (KA), an amino acid inducing limbic seizures, could elevate in the brain mRNAs encoding uPA and its specific inhibitor, plasminogen activator inhibitor-1 (PAI-1), a major antifibrinolytic agent. Brain sections encompassing the hippocampus were tested through in situ hybridization using radiolabeled riboprobes specific for the two mRNA species. The results showed that KA greatly enhanced both mRNA species in sites of limbic structures and cortex. However, in the hypothalamus and brain blood vessels only PAI-1 mRNA was elevated. Those were also the only two locations where PAI-1 mRNA was detected in the non-treated control brain, although at a low level. For both mRNAs, KA enhancement was first evident 2-4 h after treatment, and it was most prolonged in the hippocampal area, where prominent hybridization signals persisted for three days. Here, both mRNAs were initially elevated in the hilar region of the dentate gyrus and in the molecular and oriens layers; however, PAI-1 mRNA became evident throughout the area, while uPA mRNA became especially pronounced in the CA3/CA4 subfield. In the cortex both mRNA types were induced, but only uPA mRNA was elevated in the retrosplenial cortex, and also in the subiculum. In the amygdaloid complex, uPA mRNA was restricted to the basolateral nucleus, whereas PAI-1 mRNA was seen throughout the structure, however, excluding this nucleus. These data show that seizure activity enhances the expression of uPA and PAI-1 genes in the brain; the patterns of enhancement suggest that the protease and its inhibitor may act in brain plasticity in synchrony, however, also independently of each other. Furthermore, the results suggest that by elevating PAI-1 mRNA in brain blood vessels, limbic seizures generate a risk for stroke. PMID- 9221914 TI - Differential regulation of type-1 and type-2alpha corticotropin-releasing hormone receptor mRNA in the hypothalamic paraventricular nucleus of the rat. AB - Novel corticotropin-releasing hormone receptor (CRHR), designated type-2alpha CRHR (CRHR-2alpha), was recently cloned and functionally characterized. In situ hybridization study revealed that CRHR-2alpha mRNA had a distinct distribution from type-1 CRHR (CRHR-1) mRNA in the rat brain. Interestingly, CRHR-2alpha mRNA showed a relatively high expression in the hypothalamic paraventricular nucleus (PVN) even under unstressful condition. This may reflect the important role of CRHR-2alpha in the autoregulation of CRH secretion in the PVN. To determine the regulation of CRHR-2alpha mRNA expression in the PVN, we examined the alteration of CRHR-2alpha mRNA levels in the PVN in rats with lipopolysaccharide (LPS) injection, corticosterone (CORT) administration or adrenalectomy and compared with that of CRHR-1 mRNA, using in situ hybridization histochemistry. I.p. LPS injection (50 microg) induced a significant increase in PVN CRHR-1 mRNA at 3 and 6 h whereas CORT administration (10 mg/day for 12 days) or adrenalectomy (sacrificed 7 days after surgery) decreased CRHR-1 mRNA levels in the PVN. These alterations in PVN CRHR-1 mRNA are consistent with previous reports. In contrast, CRHR-2alpha mRNA levels in the PVN were not altered by any of these treatments. These results indicate that CRHR-1 and CRHR-2alpha mRNA are differentially regulated in the PVN. Further study will be necessary to elucidate the CRHR 2alpha function in the PVN. PMID- 9221915 TI - Ginsenoside Rb1 regulates ChAT, NGF and trkA mRNA expression in the rat brain. AB - Ginsenoside Rb1 (Rb1), a saponin of North American ginseng (Panax quinquefolium L.), has been found to exert beneficial effects on memory and learning, putatively through its actions on the cholinergic system. In situ hybridization studies show that Rb1 increases the expression of choline acetyltransferase and trkA mRNAs in the basal forebrain and nerve growth factor mRNA in the hippocampus. Other neurotrophins (brain-derived neurotrophic factor, neurotrophin 3), genes encoding neuropeptides (preproenkephalin, preprotachykinin) and amyloid protein precursor were also studied, but no significant change was observed. These findings support the specificity of the effects of Rb1 on certain aspects of the cholinergic and neurotrophic systems. PMID- 9221916 TI - Early induction of mRNA for calbindin-D28k and BDNF but not NT-3 in rat hippocampus after kainic acid treatment. AB - The influence of kainic acid (KA), which induces acute seizures, on expression of mRNA for the calcium-binding protein, calbindin-D28k, brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) and early-response genes [c-fos, zif268 (NGFI-A), nur77 (NGFI-B)] was examined in rat hippocampus by Northern blot analysis. A significant increase (3.2-fold) in BDNF mRNA was observed 1 h after KA injection (12 mg/kg i.p.) and peak expression (9.4-fold) occurred 3 h after KA. The induction of BDNF mRNA was preceded by the induction of c-fos, mRNA (30 min after KA) and was followed by the induction of calbindin-D28k mRNA (3.5-fold 3 h after KA; a maximal response was at 3-6 h after KA). Region-specific changes, analyzed by immunocytochemistry and in situ hybridization, indicated that the most dramatic increases in calbindin protein and mRNA after KA treatment were in the dentate gyrus. Although calbindin-D28k and BDNF mRNAs were induced, a 3.4-3.8 fold decrease in NT-3 mRNA was observed by Northern analysis 3-24 h after KA treatment. Calbindin-D28k gene expression was also examined in rats with a chronic epileptic state characterized by recurrent seizures established with an episode of electrical stimulation-induced status epilepticus (SE). When these animals were examined 30 days post-SE, no changes in hippocampal calbindin-D28k mRNA were observed. Our findings suggest that the induction of calbindin-D28k mRNA (which may be interrelated to the induction of BDNF mRNA) is an early response which may not be related to enhanced neuronal activity or seizures per se, but rather to maintaining neuronal viability. PMID- 9221917 TI - Potential effect of cytokines on transgene expression in primary fibroblasts implanted into the rat brain. AB - Fibroblasts genetically modified with retroviral vectors fail to demonstrate long term transgene expression upon implantation into the body. Although the mechanisms behind this phenomenon have not been elucidated, one likely cause is the response of the host to the graft. For example, genetically modified fibroblasts grafted into the brain are surrounded by activated microglia and astrocytes. The apparent inflammatory response can last for several weeks. In addition, the center of the graft is typically infiltrated with macrophage-like cells that appear to reside continuously within the graft. This proximity of inflammatory cells to the graft suggests that these cells may somehow influence transgene expression. In the current study, an in vitro model was used to test the effect cytokines [transforming growth factor-beta1 (TGF-beta1), interleukin 1beta, (IL-1beta) and tumor necrosis factor-alpha (TNF-alpha)] that are typically released by peripheral macrophages, activated microglia and/or astrocytes have on long-terminal repeat (LTR)-driven transgene expression in primary fibroblasts. Our data demonstrate that these cytokines can significantly reduce the steady state level of proviral mRNA. The amount of proviral mRNA returned to control levels within 24 h if the cytokines were removed. In addition, the down regulation of proviral mRNA levels could be prevented if the cells were incubated with dexamethasone (25 microM) concurrent with the introduction of cytokines. These data demonstrate that cytokines can down-regulate LTR-driven transgene expression in primary fibroblasts maintained in culture. This interaction may be a major reason why transduced cells do not demonstrate long-term transgene expression in vivo. PMID- 9221918 TI - Dynamic expression suggests multiple roles of the eph family receptor brain specific kinase (Bsk) during mouse neurogenesis. AB - The eph family ligands and receptors have been implicated in mediating topographic neuron-target interactions. We recently isolated Bsk, a new member of the eph family receptors, and showed that it is expressed primarily in the brain. To investigate the role of Bsk in the development of the nervous system, we examined the temporal and spatial patterns of Bsk expression using in situ hybridization. We report here that Bsk expression exhibits dynamic changes during embryogenesis. In early embryos, Bsk is widely transcribed in the nervous system, including the forebrain, midbrain, hindbrain and spinal cord. Bsk expression in the midbrain, hindbrain and spinal cord, however, gradually decreases while in the forebrain increases over time. By embryonic day 18, the most intense Bsk expression was found in the limbic system. High levels of the expression in the limbic system persisted throughout post-natal development and remained stable in the adult up to 24 month. The topography of Bsk expression is in the form of gradients in several regions of the brain, including the lateral septum, spinal cord, as well as the hippocampus. Selective expression was also observed in Purkinje cells. Our findings on the topography of Bsk expression provide support to potential roles of Bsk in topographic projection. Our analyses further suggest that there may be other novel functions of Bsk in early neurogenesis in addition to potential roles in topographic mapping. PMID- 9221920 TI - End-capped antisense oligodeoxynucleotides effectively inhibit gene expression in vivo and offer a low-toxicity alternative to fully modified phosphorothioate oligodeoxynucleotides. AB - Sulfur modification of oligodeoxynucleotides produces nuclease resistance but also leads to toxic effects when these compounds are administered in vivo. To assess their potential as viable alternatives to full phosphorothioate derivatives, we have used rotational behavior and immunohistochemistry to investigate the efficacy and longevity of phosphorothioate, end-capped antisense oligodeoxynucleotides in suppression of c-fos and ngfi-a in the striatum of adult rats. Our results suggest that, despite having a limited duration of action, these end-capped, chimeric oligodeoxynucleotides are capable of specifically inhibiting gene expression in vivo and may, therefore, possess broader application potential in chronic suppression models as the reduction of sulfur content is likely to greatly minimize their toxic effects. PMID- 9221919 TI - Alterations in the estrogen sensitivity of hypothalamic proenkephalin mRNA expression with age and prenatal exposure to alcohol. AB - Studies suggest that exposure to alcohol in utero causes reproductive and neuroendocrine deficits in adult female rats. The ventromedial nucleus of the hypothalamus (VMN) is an estrogen-sensitive brain region which is regarded as a primary locus for modulating female reproduction. Proenkephalin (PE) mRNA expression in the VMN is dramatically increased by estrogen and this elevation is thought to be involved in modulating female reproductive behavior and neuroendocrine function. To examine whether prenatal alcohol exposure has long term effects on the ability of estrogen to influence hypothalamic PE mRNA levels, female rats at 2-3, 6-7 or 15-18 months of age, derived from alcohol- or control fed dams, were studied. 7 days following ovariectomy, animals received either estrogen or sham treatment for 2 days prior to sacrifice. PE mRNA levels in the VMN and striatum were determined by in situ hybridization histochemistry. Film autoradiogram density, numbers of PE mRNA-expressing cells and exposed silver grains/cell were analyzed. Estrogen treatment increased hybridization density, the number of PE mRNA-expressing cells and PE mRNA (grains) level/cell in the VMN of normal adult female rats. In old rats, estrogen increased the number of PE mRNA-expressing cells without up-regulating PE mRNA grain density/cell. In fetal alcohol-exposed (FAE) female rats, the number of cells that expressed PE mRNA did not increase following estrogen treatment at any age. Elevation of grain density/cell following estrogen was observed in FAE animals but only at 7-8 months of age. Overall, these data indicate that the estrogen responsiveness of PE mRNA expression in the VMN declines with age and, furthermore, prenatal exposure to alcohol blunts estrogen's effects on PE mRNA expression in the adult VMN. These finding may help to explain the mechanisms underlying the loss of reproductive function observed in FAE females. PMID- 9221921 TI - Modulation of angiotensin II type 2 receptor mRNA in rat hypothalamus and brainstem neuronal cultures by growth factors. AB - This study investigates the regulatory effects of growth factors upon angiotensin II type 2 (AT2) mRNA levels in neurons co-cultured from newborn rat hypothalamus and brainstem. Incubation of cultured neurons with nerve growth factor (NGF; 5-50 ng/ml) caused time-dependent changes in the steady-state levels of AT2 receptor mRNA. Short-term (0.5-1.0 h) incubations with NGF resulted in significant increases in AT2 receptor mRNA, whereas longer-term incubations (4-24 h) caused significant decreases. Activation of NGF receptors is known to stimulate phospholipase C-gamma and subsequently activate protein kinase C (PKC). Incubation of cultures with the PKC activator, phorbol-12-myristate-13-acetate (PMA; 100 nM), caused temporal changes in AT2 receptor mRNA levels similar to those observed with NGF. By contrast, insulin (0.1-10 microg/ml) elicited only significant decreases in AT2 receptor mRNA levels. The observed abilities of NGF and insulin to regulate the expression of AT2 receptor mRNA are consistent with the fact that the AT2 receptor gene promoter region contains several cis DNA regulatory elements that respond to growth factor-stimulated transcription factors. These novel observations which show that NGF and insulin can regulate AT2 receptor mRNA in neurons derived from neonatal rat CNS lend support to the idea that AT2 receptors have a role in development and differentiation. PMID- 9221922 TI - A comparison of changes in nucleotide-protein interactions in the striatal, hippocampus and paramedian cortex after cerebral ischemia and reperfusion: correlations to regional vulnerability. AB - [32P]Azido-purine analogs of ATP and GTP were used to detect changes in phosphorylation and nucleotide binding induced by ischemia and subsequent reperfusion in rat brain striatum, hippocampus and paramedian cortex (PM cortex) tissues. Major changes in phosphorylation were observed for a 130-kDa protein, tentatively identified as the Ca2+ transport ATPase, and calcium/calmodulin dependent protein kinase II (CaM Kinase II) in all tissues. However, recovery of the phosphorylation of the 130-kDa protein occurred only in the PM cortex on reperfusion. A 200-300% increase in [32P]8N3ATP photoinsertions was observed in the striatum and hippocampus regions for a 43-kDa protein with an isoelectric point of 6.8. This protein was identified as glutamine synthetase (GS) and the increase in binding was found to be due to both increased copy number and activation by Mn2+. An increase in [32P]8N3GTP photoinsertion into a 55-kDa protein, identified as the beta-subunit of tubulin, was found only in the striatum and hippocampus. This indicates the depolymerization of microtubulin in these tissues. These changes correlate to the vulnerability of the striatum and hippocampus to ischemia-induced neuronal death. PMID- 9221923 TI - Rat nurr1 is prominently expressed in perirhinal cortex, and differentially induced in the hippocampal dentate gyrus by electroconvulsive vs. kindled seizures. AB - We isolated a rat orphan nuclear hormone receptor from a brain cortex cDNA library. The sequence of the cDNA insert was 2154 bp with an open reading frame of 1794 bp encoding a putative protein of 598 amino acids and predicted molecular mass of 65 kDa. The deduced amino acid sequence showed a strong homology to the mouse nurr1 and human NOT1 orphan nuclear hormone receptors of the NGFI B/nur77/NAK1 gene subfamily. We refer to this rat clone as r-nurr1. Northern blot analysis showed that r-nurr1 mRNA was highly expressed in the brain and moderately in the lung as a 4.0 kb transcript. A smaller transcript of 2.5 kb was also detected in the testes. The level of r-nurr1 transcript in the heart, skeletal muscle, liver, kidney and spleen was marginal. In situ hybridization showed that r-nurr1 mRNA was constitutively expressed in various regions of the CNS, particularly in the deeper layers (IV to VI) of the perirhinal cortex and area 2 of parietal cortex. We further evaluated the modulation of r-nurr1 expression in CNS by an electroconvulsive seizure (ECS) and by an amgydala kindled seizure. A single ECS administered via earclip electrodes induced a rapid and transient increase of r-nurr1 mRNA in the granule cells of the dentate gyrus, being significant at 15 min after the seizure, maximal approximately 1 h and back to baseline at 4 h. The amygdala kindled seizure revealed a less robust and restricted nurr-1 induction in the CNS, as only two of the four kindled animals showed a unilateral induction of nurr1 mRNA in the dentate gyrus. These results suggest that r-nurr1 is an immediate-early gene that is differentially induced by ECS vs. kindled seizures. In addition, as r-nurr1 is prominently expressed in the specific brain sites associated with memory acquisition and consolidation, it may play a role in memory processing. PMID- 9221924 TI - Transient expression of FGF-5 mRNA in the rat cerebellar cortex during post-natal development. AB - Previously, we showed that fibroblast growth factor (FGF) receptor-4 mRNA was transiently expressed in proliferative granule cells of the external granule layer of the rat cerebellar cortex during early post-natal development (A. Miyake et al., Mol. Brain Res., 31 (1995) 95-100). In this study, we examined the expression of FGF-5 mRNA in the rat brain during post-natal development by in situ hybridization. FGF-5 mRNA was transiently expressed in granule cells of the internal granule layer of the cerebellar cortex during early post-natal development. The temporal sequence of FGF-5 mRNA expression was similar to that of FGFR-4 mRNA expression. As the proliferation of granule cells in the external granule layer and their migration through the molecular layer into the internal granule layer actively occur during these periods, the present findings suggest that FGF-5 as well as FGFR-4 might play important roles in the proliferation and/or migration of granule cells during the post-natal development of the cerebellar cortex. PMID- 9221925 TI - Reduced expression of alpha2C-adrenoceptors in rat striatum following antisense oligodeoxynucleotide infusion. AB - The predominate subtypes of alpha2-adrenoceptors in the brain are alpha2A and alpha2C. The lack of selective ligands for these receptors hampers their functional characterization. We exploited an antisense strategy as an alternative pharmacological tool to study alpha2C-adrenoceptors. In rat striatum (caudate putamen), alpha2-adrenoceptors were characterized using the subtype-non-selective antagonist [3H]2-(2-methoxy-1,4-benzodioxan-2-yl)-2-imidazoline ([3H]RX821002). Specific [3H]RX821002 binding was saturable and to a single class of high affinity sites. Curves for the inhibition of [3H]RX821002 binding by the alpha2C selective compound, prazosin, were fit best by a model assuming binding to two sites, presumably reflecting binding to alpha2A- and alpha2C-adrenoceptors. A 15 mer phosphorothioate oligodeoxynucleotide (alpha2C AS) complementary to the alpha2C-adrenoceptor mRNA, or a random sequence (RS) was administered to rats continuously for 4.5 days directly into the striatum. Compared to RS infusions, alpha2C AS infusions induced a 35% reduction in the Bmax of [3H]RX821002 in striatal homogenates (P < 0.05). Curves for the inhibition of [3H]RX821002 binding by prazosin were fit best by a model assuming a single interaction in alpha2C AS-infused rats and to a model assuming two sites in RS-infused rats. These results are consistent with the conjecture that both alpha2A- and alpha2C adrenoceptors occur in the rat striatum and also demonstrate the feasibility of an antisense approach to examine the functional role of subtypes of alpha2C adrenoceptors in the brain. PMID- 9221926 TI - Haloperidol regulates neurotensin gene expression in striatum of c-fos-deficient mice. AB - The immediate-early gene c-fos has been proposed to play a role in induction of neurotensin/neuromedin N (NT/N) gene expression in the striatum following acute haloperidol (HAL) treatment. We utilized mice with targeted disruption of the c fos gene to directly test this hypothesis. A robust increase in NT/N gene expression was observed in the dorsolateral striatum (DLSt) in both wild-type (WT) and c-fos-deficient mice 4-6 h after a single injection of HAL (1 or 4 mg/kg) indicating that products of the c-fos gene are not absolutely required for induction of NT/N mRNA. The basal expression of preprotachykinin, preproenkephalin and preprocholecystokinin mRNAs did not differ between WT and c fos knockout mice. HAL treatment first increased striatal NT/N mRNA on postnatal day (PD) 10. HAL-induced NT/N mRNA levels were significantly lower in c-fos knockout mice than in WT mice on PD 10 and 15. These findings indicate that reliance on c-fos may be greater earlier in development and that redundant molecular pathways can lead to induction of NT/N mRNA in mouse striatum. PMID- 9221927 TI - AMPA receptor subunits expressed by single astrocytes in the juvenile mouse hippocampus. AB - The subunit composition of native AMPA receptor (AMPA-R) channels was recently described in several neuronal cell types but less information is available on glial cells. Evidence from recombinant receptor studies suggests that the expression of distinct subunits determines the specific functional properties of the receptor channel. In the present study, we combined the patch clamp technique with the reverse transcription-polymerase chain reaction (RT-PCR) to correlate the expression of gene transcripts with functional properties of AMPA-R in single identified glial cells of the hippocampus. The cells were freshly isolated from the stratum radiatum of the CA1 subregion. We focused on cells expressing AMPA-R with an intermediate Ca2+ permeability which were identified as immature astrocytes due to their morphological, immunocytochemical and electrophysiological characteristics. After recording, the cells were harvested and RT-PCR was performed with the same individual cell to investigate the composition of their AMPA-R transcripts. Our results suggest the expression of a heteromeric subunit architecture. In all cells, the GluR2 subunit was present, which is known to confer a low Ca2+ permeability to the receptor complex. Most frequently, we met co-expression of GluR2 and GluR4. This study demonstrates that astrocytes in the hippocampus express a distinct AMPA-R subunit composition which differs from neurons. The glial receptors might be involved in the modulation of gene expression as well as the regulation of proliferation and differentiation. PMID- 9221928 TI - Characterization of brain ecto-apyrase: evidence for only one ecto-apyrase (CD39) gene. AB - A rat brain cDNA coding for ecto-(Ca,Mg)-apyrase activity was isolated using human CD39 cDNA and functionally expressed in COS-7 cells. The gene codes for a protein with high similarity to human (75% identity) and murine (90% identity) CD39. It is expressed in primary neurons and astrocytes in cell culture as well as in kidney, liver, muscle and spleen. Southern analysis of the mouse genome suggests that there may be a single copy of the ecto-apyrase gene. Interestingly, the human CD39 gene cytologically co-localizes with the susceptibility gene involved in human partial epilepsy with audiogenic symptoms; such a coincidence is consistent with reports on the deficiency of ecto-apyrase activity in the brains of humans with temporal lobe epilepsy and in those of mice with audiogenic seizures. PMID- 9221929 TI - The effect of cycloheximide on the regulation of proenkephalin and prodynorphin gene expressions induced by kainic acid in rat hippocampus. AB - The effect of cycloheximide (CHX), a protein synthesis inhibitor, on the regulation of proenkephalin (proENK) and prodynorphin (proDYN) mRNA levels, proto oncogenes, such as c-fos, 35-kDa fra and c-jun mRNA, and the levels of their products induced by kainic acid (KA) in rat hippocampus was studied. The proENK and proDYN mRNA levels were markedly increased 4 and 8 h after KA (10 mg/kg i.p.) administration. However, the intracellular proENK protein level was not affected by KA. The elevations of both proENK and proDYN mRNA levels induced by KA were inhibited by pre-administration of CHX (15 mg/kg i.p.). The increases of proENK and proDYN mRNA levels induced by KA were well-correlated with the increases of c Fos, 35-kDa Fra and c-Jun protein levels. KA administration increased the hippocampal levels of c-Fos, 35-kDa Fra and c-Jun proteins with the time. The increases of c-Fos, 35-kDa Fra and c-Jun protein levels induced by KA administration were also inhibited by CHX pre-administration. KA administration markedly increased both c-fos and c-jun mRNA levels during 1 and 4 h and the increased levels of these proto-oncogene mRNA were further prolonged by the treatment with CHX. In addition, CHX alone increased both c-fos and c-jun mRNA levels although the onset times of induction were different. In electrophoretic mobility shift-assay, both AP-1 and ENKCRE-2 DNA-binding activities were increased by KA. KA-induced increases of AP-1 and ENKCRE-2 DNA-binding activities were also attenuated by CHX. In addition, KA-induced AP-1 and ENKCRE-2 DNA binding activities were diminished by the antibodies against Fos and Jun family proteins. Furthermore, the cross-competition studies revealed that AP-1 proteins actively participated in ENKCRE-2 DNA domain. The results suggest that KA-induced proENK and proDYN mRNA expressions may require on-going synthesis of proteins, such as c-Fos, c-Jun and 35-kDa Fra, which may have a possible role in the up regulation of proENK and proDYN gene expression through the binding with AP-1 and ENKCRE-2 DNA-binding motifs. PMID- 9221930 TI - Differential regional effects of long-term L-DOPA treatment on preproenkephalin and preprotachykinin gene expression in the striatum of 6-hydroxydopamine lesioned rat. AB - The present study examined the effects of prolonged L-DOPA treatment (6 months) alone or in combination with unilateral 6-hydroxydopamine-induced lesion of the mesostriatal dopaminergic pathway on substance P and enkephalin mRNA expression in the rat neostriatum. This was done by means of quantitative in situ hybridization histochemistry. As reported previously, the unilateral dopaminergic lesion induced a significant and homogeneous decrease in striatal substance P mRNA expression and a marked increase in enkephalin mRNA expression in the ipsilateral neostriatum which was more pronounced in the dorsolateral than ventromedial part of the structure. Long-term L-DOPA treatment alone had no significant effects on the two striatal peptide mRNA levels. The chronic L-DOPA treatment in 6-hydroxydopamine-lesioned rats was found to partially reverse the lesion-induced down-regulation of substance P mRNA expression, without significantly affect the up-regulation of enkephalin when considering the neostriatum as a whole. Topographical analysis revealed that long-term L-DOPA treatment reversed, in fact, both post-lesional enkephalin and substance P responses to 6-hydroxydopamine lesion, in the ventromedial neostriatum, without significantly modified these peptide responses in the dorsolateral neostriatum. These findings provide new evidence that prolonged L-DOPA treatment differentially affects the post-lesional peptide responses in the ventromedial and dorsolateral parts of the neostriatum, suggesting regional cellular mechanisms in the neostriatum underlying the benefit and/or side-effects of L DOPA treatment in parkinsonian patients. PMID- 9221931 TI - Islet amyloid polypeptide and calcitonin gene-related peptide expression are down regulated in dorsal root ganglia upon sciatic nerve transection. AB - Islet amyloid polypeptide (IAPP) is structurally related to calcitonin gene related peptide (CGRP) and has been implicated in glucose homeostasis and diabetes pathogenesis because it is expressed in insulin cells and forms amyloid in pancreatic islets from type II diabetic patients. IAPP is also constitutively co-expressed with CGRP in rat sensory neurons. Whether expression of IAPP is altered by nerve injury with or without regeneration was investigated in adult rats subjected to unilateral sciatic axotomy; IAPP and CGRP expression were determined by quantitative in situ hybridization and immunocytochemistry at days 3, 10 and 30 after axotomy. In ipsilateral L4-L5 dorsal root ganglia (DRG), the percentages of nerve cell profiles labelled for IAPP and CGRP mRNA were reduced at all time points studied. IAPP and CGRP mRNA expression were lower in nerve cell profiles in ipsilateral DRGs compared to the contralateral side after axotomy alone whereas epineurial nerve suture maintained or restored IAPP and CGRP expression. The numbers of IAPP- and CGRP-immunoreactive DRG nerve cell profiles and dorsal horn fibers were reduced on the ipsilateral side at all time points. Thus, IAPP and CGRP expression are down-regulated upon axotomy. Nerve repair maintains or restores IAPP and CGRP expression in individual neurons but does not prevent the loss of CGRP/IAPP phenotype of some of these neurons in response to axotomy. PMID- 9221932 TI - Clozapine and haloperidol differentially affect AMPA and kainate receptor subunit mRNA levels in rat cortex and striatum. AB - Dopamine is the neurotransmitter most often implicated in the pathogenesis of schizophrenia. However, glutamatergic antagonists can cause psychotic symptoms in otherwise normal humans, and exacerbate these symptoms in schizophrenics. These findings have led to a model of dopamine-glutamate interactions in limbic cortex and striatum as a potential substrate for symptom production in schizophrenia. From this model, we might expect that cortical and striatal expression of non NMDA ionotropic glutamate receptors would be differentially regulated by antipsychotic treatment. To begin to address this question, we examined the regulation of mRNA levels of the AMPA (gluR1-gluR4), low affinity kainate (gluR5 gluR7), and high affinity kainate (KA1-KA2) receptor subunits by clozapine (20 mg/kg/day) and haloperidol (2 mg/kg/day) treatment for 2 weeks. Both clozapine and haloperidol caused region-specific alterations in the mRNA levels of these subunits, but there was no differential regulation in the cortex vs. the striatum. Haloperidol caused a decrease in gluR2 and gluR4 mRNA levels in both cortex and striatum and an increase in KA2 mRNA levels in the striatum only. However, clozapine treatment caused an increase in gluR7 mRNA expression, and a decrease in gluR3 mRNA expression, in both cortex and striatum while causing an increase in KA2 mRNA levels, and a decrease in gluR4 mRNA levels, in the striatum only. These dissimilarities may represent an interesting mechanism for some of the differential therapeutic or toxic effects of clozapine and haloperidol, and also may be relevant to our understanding of dopamine-glutamate interactions in schizophrenia. PMID- 9221933 TI - Arginine-481 mutation abolishes ligand-binding of the AMPA-selective glutamate receptor channel alpha1-subunit. AB - Arginine-481 is located in the putative agonist-binding region preceding the putative transmembrane segment M1 of the alpha1-subunit of the AMPA-selective glutamate receptor (GluR) channel. This amino acid is completely conserved among GluR proteins. A site-directed mutagenesis study using a baculovirus expression system showed that substitution of glutamate, glutamine and lysine for arginine 481 of the recombinant alpha1-subunit protein abolishes binding to [3H]AMPA completely. The present study provides the first direct experimental evidence that the conserved charged arginine-481 residue is essential, directly or indirectly, for the acquisition of ligand-binding activity by the receptor protein. PMID- 9221934 TI - A novel splice variant of the cell adhesion molecule BIG-2 is expressed in the olfactory and vomeronasal neuroepithelia. AB - We have cloned a mouse cDNA encoding a novel truncated form of the gene BIG-2 from the vomeronasal organ. The related proteins BIG-2 and BIG-1 possess a C terminal glycosylphosphatidylinositol anchor, six immunoglobulin domains and four fibronectin type III repeats. They are related to certain axonal-associated cell adhesion molecules (AxCAMs) exhibiting most similarity to the TAG-1/F3 subgroup of neural cell adhesion molecules. The cDNA we have identified, termed BIG-2A, appears to represent a novel splice variant of BIG-2 possessing six Ig-like domains, a single fibronectin repeat and lacking the glycosylphosphatidylinositol anchoring domain (GPI). To determine the expression of this gene, in situ hybridization analysis was performed in adult and developing mice using a riboprobe specific for BIG-2A. Maximum expression was observed in mature sensory cells of the vomeronasal neuroepithelium and a less intense signal was also evident in the olfactory neuroepithelium. These results suggest that alternative splicing of the BIG-2 gene transcript may play an important role in the organization of the vomeronasal and olfactory neuroepithelia. PMID- 9221936 TI - Increase of interleukin-1beta mRNA and protein in the spinal cord following experimental traumatic injury in the rat. AB - Interleukin-1 beta (IL-1beta) is a major mediator of inflammation and a growth promoter for many cell types that could play an important role in the consequences of traumatic spinal cord injury (SCI). In the present study, the expression of IL-1beta and its mRNA was determined in the rat spinal cord following a standardized contusion injury. IL-1beta mRNA, measured with quantitative RT-PCR, was significantly increased in the lesion site by 1 h after SCI (35.2 +/- 5.9 vs. 9.1 +/- 2.1 pg/mg RNA, n = 3, P < 0.05) and remained significantly higher than in the normal spinal cord for at least 72 h post-injury (p.i.). IL-1beta mRNA levels in tissue immediately caudal to the lesion site did not change after the injury. IL-1beta protein levels, measured by an ELISA, were determined at the lesion site and in cerebrospinal fluid (CSF) and serum samples. IL-1beta levels in the CSF and serum were much lower than in the spinal cord. At the lesion site, IL-1beta was increased significantly by 1 h p.i., peaked at 8 h (32.3 +/- 0.1 vs. 7.6 +/- 1.9, ng/g tissue, n = 5, P < 0.05) and remained significantly higher than normal through at least 7 days p.i. These results suggest that the increased IL-1beta mRNA and protein levels are an early and local response at the lesion site that could trigger other, later, responses to traumatic SCI. PMID- 9221935 TI - GABA(A) and GABA(B) agonists and antagonists alter the phase-shifting effects of light when microinjected into the suprachiasmatic region. AB - GABAergic drugs have profound effects on the regulation of circadian rhythms. The present study evaluated the effects of microinjections of GABAergic drugs into the suprachiasmatic region in hamsters on phase shifts induced by light and by microinjection of a cocktail containing vasoactive intestinal peptide (VIP), peptide histidine isoleucine (PHI) and gastrin-releasing peptide (GRP). The phase advancing effects of light at circadian time (CT) 19 were significantly reduced by microinjection of GABA(A) or GABA(B) agonists into the SCN, but were not altered by microinjection of GABA(A) or GABA(B) antagonists. Microinjection of a GABA(B) agonist also reduced the phase-delaying effects of light at CT 13.5-14 while a GABA(B) antagonist increased the phase delays caused by light. Neither GABA(B) drug altered the phase delays produced by microinjection of a peptide cocktail containing VIP, PHI, GRP. These data indicate that changes in GABA(A) or GABA(B) activity within the SCN can alter the phase-shifting effects of light on circadian rhythms and support a role for GABA in gating photic input to the circadian clock. PMID- 9221937 TI - Nucleus raphe obscurus (nRO) regulation of anorectal motility in rats. AB - Previous research has demonstrated that anorectal contractions in the rat are modulated by activation of spinal autonomic circuits. In the present study, anterograde tracing of descending pathways originating from the caudal nucleus raphe obscurus (nRO) revealed that this nucleus projects to cells within the intermediolateral (IML) cell column of the thoracic cord and the sacral parasympathetic nucleus (SPN). These anatomical studies suggested that the nRO may influence the regulation of spinal reflexes of the pelvic floor. In a second set of experiments, acute rat preparations were used to investigate changes in anorectal motility during electrical stimulation of the nRO. Anorectal contractions were measured by a fluid-filled manometer. Electrical stimulation of the nRO significantly reduced spontaneous anorectal activity when compared to baseline contractions recorded for 1 min prior to stimulation. Stimulation sites outside the nRO did not affect anorectal contractions when compared to either (a) the 1-min pre-stimulation baseline for that site or (b) the 1-min stimulation period for sites within the nRO. Stimulation of caudal portions of the nRO were more likely than the rostral nRO to reduce anorectal contractions. Given that the SPN contains preganglionic neurons which may be involved in control of anorectal contractions (mediated via the pelvic nerve), the studies presented here suggest a functional role for nRO regulation of preganglionic motoneurons innervating the distal gut of the rat. PMID- 9221938 TI - Differential effects of single and repeated ketamine administration on dopamine, serotonin and GABA transmission in rat medial prefrontal cortex. AB - Cognitive functions regulated by the prefrontal cortex are sensitive to changes in dopaminergic and serotoninergic transmission. The non-competitive N-methyl-D aspartate (NMDA) receptor antagonist ketamine influences dopaminergic transmission and induces psychotic symptoms in normal and schizophrenic individuals. This study examined the effect of single and repeated ketamine (25 mg/kg, i.p.) administration on extracellular levels of dopamine, GABA and the serotonin metabolite 5-hydroxyindoleacetic (5-HIAA) acid in the medial prefrontal cortex using in vivo microdialysis in conscious rat. In line with earlier studies, we observed a transient five-fold increase in dopamine release following single ketamine administration in drug naive animals. However, we also observed a two-fold increase in basal dopamine levels and an almost complete attenuation of the ketamine-induced increase in dopamine release in animals pre-treated with ketamine once daily for 7 days. Extracellular 5-HIAA levels were increased by ketamine in both drug naive and even more enhanced in ketamine-pre-treated animals but without a change in basal 5-HIAA levels. GABA levels were unaffected by either single or repeated ketamine administration. We demonstrate evidence for a differential effect of single and repeated ketamine administration on dopamine, serotonin and GABA transmission in the medial prefrontal cortex. We provide new evidence for a complex adaptation of neurotransmission following repeated NMDA receptor blockade whereby in the presence of increased basal dopamine levels the ketamine-induced increase in dopamine is attenuated and the increase in 5-HIAA is enhanced. It appears from our results that ketamine pre-treatment reduces the dynamics of dopaminergic transmission in the prefrontal cortex and may possibly alter the balance between dopamine and serotonin transmission. PMID- 9221939 TI - Glucocorticoids modulate G-protein alpha-subunit levels in PC12 cells. AB - Regulation by the synthetic glucocorticoid hormone, dexamethasone, of the levels of several G-protein alpha-subunits was studied during differentiation in PC12 cells. Similar patterns, although with different magnitudes, were observed in the changes in the levels of alpha il, alpha s, and alpha q induced by the treatments studied, whereas alpha o differed from the other alpha-subunits. Thus, nerve growth factor (NGF) treatment increased alpha il, alpha s, and alpha q, and forskolin increased alpha il and alpha q, with the increase in alpha il being greater than the increases in the other two alpha-subunits after both treatments. The increases in alpha il, alpha s, and alpha q induced by NGF were dependent on signaling through ras, since they did not occur in NGF-treated M17 cells, which express a dominant inhibitory Ha-ras. Treatment of PC12 cells with dexamethasone antagonized the increases in alpha il, alpha s, and alpha q induced by NGF or forskolin, almost completely blocking any changes from control levels. The level of alpha o also was increased in PC12 cells by treatment with NGF or forskolin, but, in contrast to the other G-protein alpha-subunits, the response to NGF was not antagonized by dexamethasone in PC12 cells, or by the deficient ras activity in M17 cells. However, ras influenced the alternative splicing that regulates the levels of the two alpha o subtypes, beta o1 and alpha o2, so they were expressed in a ratio of 1:2 in PC12 cells but 2:1 in ras-deficient M17 cells. These results demonstrated marked, and subtype-selective, influences of dexamethasone on the levels of G-protein alpha-subunits, an effect that may contribute to the effects of conditions that increase the levels of glucocorticoid hormones, such as stress or certain diseases, on signal transduction processes in brain. PMID- 9221940 TI - Extracellular glutamate and dopamine measured by microdialysis in the rat striatum during blockade of synaptic transmission in anesthetized and awake rats. AB - We investigated the effect of high dose tetrodotoxin (TTX) on microdialysis measurements of extracellular striatal glutamate and dopamine in normal female rats. Both halothane-anesthetized rats with acutely implanted microdialysis probes and awake rats with microdialysis probes implanted for 24 h were tested. Glutamate levels in awake rats were 45% higher than those of anesthetized rats. Extracellular glutamate remained TTX-insensitive regardless of TTX concentration, anesthesia, or time lapsed after probe implantation. In contrast, TTX reduced dialysate dopamine in all TTX concentrations tested. We speculate that the lower glutamate levels in anesthetized rats reflect the effect of anesthesia. Because glutamate is involved, either as a reactant or a product in a variety of reactions critical to intermediary metabolism in the brain, basal dialysate glutamate levels might indirectly reflect brain metabolism. Further, we conclude that extracellular glutamate collected during non-stimulated conditions is TTX insensitive. The fact that glutamate levels are TTX-independent does not rule out that glutamate is synaptic in origin but rather demonstrates that it is not nerve impulse-dependent. However, the brain interstitial glutamate pool accessible to the microdialysis probe during control conditions is most likely isolated from the synapse, and therefore does not impose a neurotoxic potential. PMID- 9221941 TI - Susceptibility to apoptosis is enhanced in immature cortical neurons. AB - The susceptibility of cortical neurons to two forms of apoptotic death was compared with susceptibility to excitotoxic death during development in vitro (DIV 4-21). Murine cortical cultures were exposed for 48 h to the phosphatase inhibitor cyclosporine, the protein kinase inhibitor staurosporine or the excitotoxin N-methyl-D-aspartate (NMDA). Susceptibility to apoptosis induced by staurosporine or cyclosporine was maximal between DIV 4-10 and declined from DIV 10 through 18. The opposite pattern was observed with susceptibility to NMDA receptor-mediated excitotoxic necrosis, which was minimal at DIV 6 and progressively increased through DIV 21. PMID- 9221942 TI - Effects of partial nerve injury on the responses of C-fiber polymodal nociceptors to adrenergic agonists. AB - The effects of partial division of the great auricular nerve of adult rabbits were evaluated on the responsiveness of cutaneous C-fiber polymodal nociceptors (CPMs) to sympathetic stimulation (SS), close-arterial injections of epinephrine (EPI) and other alpha-adrenergic agonists. In normal unanesthetized rabbits, the two ears were usually at the same temperature. Two to 4 weeks after partial nerve lesions, however, the operated ear was cooler by 1-3 degrees C in the majority of animals, suggestive of increased vasoconstriction and possible denervation supersensitivity. Neither SS nor EPI (50 ng) excited CPM units (n = 23) from intact anesthetized animals. In contrast, 14-27 days after partial nerve lesions, SS (8 out of 38 units) and EPI (12 out of 38 units) were excitatory for a class of CPMs. There was notable variability in the response of different units and of a given unit between first and second trials. Responses consisted of 1-22 impulses for SS and 1-23 impulses for EPI in the 60 s following a trial. Arterial occlusion did not activate responsive units, suggesting that the excitation was not caused by vascular or temperature changes. Selective alpha2-adrenoceptor blockade with yohimbine (0.6-1.0 mg/kg i.v.) or rauwolscine (1.0 mg/kg i.v.) reversibly antagonized the effects of SS and EPI. EPI-responsive units were also excited by norepinephrine (50 ng) and guanabenz (10 microg) but not by clonidine (3 microg) or B-HT 933 (3 microg). The results suggest that circulating EPI, acting via an alpha-adrenoceptor subtype, can play a part in the development and/or maintenance of aberrant pain syndromes such as causalgia and other sympathetically related dystrophies. PMID- 9221943 TI - Antinociceptive involvement of substance P in the spinal cord of mice: dose effects of substance P on the behavior elicited by intrathecally administered NMDA. AB - The functional interaction between substance P (SP) and N-methyl-D-aspartate (NMDA) was studied to clarify the diversity of the roles of SP in nociceptive processes at the spinal level in mice. Behavioral responses elicited by intrathecal co-administration of NMDA (0.25 nmol) with various doses of SP (0.3 12 pmol) were observed for 1 min. The high dose of SP (12 pmol) potentiated NMDA induced responses, which consisted of caudally directed licking and biting, while the low dose of SP (1 pmol) significantly reduced the responses by 40% compared to control mice administered NMDA alone. The antinociceptive effect of the low dose of SP was negated by co-administration of the opioid receptor antagonist naloxone. Furthermore, the antinociception produced by SP was present in mice pre treated with systemic administration of capsaicin during the neonatal period. These results suggest that one of the roles SP plays at the spinal level is an involvement in antinociception. The activities of excitatory dorsal horn neurons are considered to be inhibited by endogenous opioid peptides released from inhibitory dorsal horn neurons directly stimulated by SP. PMID- 9221944 TI - Attenuation of nitric oxide synthase induction in IRF-1-deficient glial cells. AB - Nitric oxide (NO) produced by inducible nitric oxide synthase (iNOS) exerts inhibitory and cytotoxic effects on various cells including neuronal cells. Glial NO production, mediated via induction of iNOS, is thought to facilitate neuronal damage during cerebral ischemia. Recently, interferon regulatory factor-1 (IRF-1) has been reported to be an essential transcription factor for iNOS mRNA induction in murine macrophages. However, expression of IRF-1 and its role in the central nervous system have not been examined. In the present study, by using primary glial cell cultures from mice with targeted disruption of the IRF-1 gene, we investigated whether IRF-1 is involved in iNOS mRNA induction in glial cells. After stimulation with lipopolysaccharide and interferon-gamma, IRF-1 mRNA was strongly induced in wild-type (IRF-1 +/+) glial cells. iNOS mRNA induction and nitrite production in IRF-1 -/- glial cells were reduced as compared with those observed in IRF-1 +/+ glial cells. Diethyldithiocarbamate, a selective inhibitor of nuclear transcription factor kappa B (NF-kappa B), completely inhibited iNOS mRNA induction. These results suggest that not only NF-kappa B but also IRF-1 play important roles in iNOS mRNA induction in the central nervous system. PMID- 9221945 TI - Burst activity of ventral tegmental dopamine neurons is elicited by sensory stimuli in the awake cat. AB - In light of evidence implicating dopamine in the pathophysiology of attention deficit disorder and schizophrenia, diseases involving attentional or sensory processing abnormalities, it was of interest to determine whether and how dopamine neurons in the ventral tegmental area respond to sensory stimuli. The single-unit responses of ventral tegmental dopamine neurons were recorded in freely-moving cats during the presentation of brief, non-conditioned auditory and visual stimuli. Both auditory and visual stimuli produced neuronal excitation, involving a greater than 5-fold increase in the probability of burst firing followed by a period of burst inhibition. The burst nature of the single-unit response suggests that sensory-induced dopamine release at target sites was disproportionally large relative to the discharge frequency. While characteristics of the dopaminergic sensory response were similar for auditory and visual stimuli, the response latency was longer for visual stimuli. The results demonstrate that dopamine neurons in the ventral tegmental area, the site of origin for mesolimbocortical dopamine neurons, are reliably activated by non conditioned auditory and visual stimuli. PMID- 9221946 TI - Nicotine activates NPY and catecholaminergic neurons in brainstem regions involved in ACTH secretion. AB - Nicotine rapidly and potently stimulates ACTH secretion via a centrally mediated mechanism. The purpose of the current study was to identify the phenotype of nicotine-sensitive neurons in brainstem catecholaminergic regions previously shown to be responsive to nicotine. Immunocytochemical double-labeling was used to detect c-Fos expression in neurons positive for activin, galanin, or neuropeptide Y (NPY), in comparison to those containing tyrosine hydroxylase (TH, catecholaminergic biosynthetic enzyme). These neuropeptides were chosen because (1) each is located in nicotine-sensitive brainstem regions, (2) neurons containing each of these peptides project to the hypothalamic paraventricular nucleus, and (3) each has been shown to affect ACTH secretion. Freely moving, adult, male rats received an intravenous (i.v.) infusion of saline or nicotine (0.045 mg/kg over 30 s or 0.135 mg/kg over 90 s) and were cardiac perfused 60 min thereafter. Nicotine significantly increased c-Fos expression in a dose-dependent manner in the brainstem regions examined. In nucleus tractus solitarius (NTS)-A2 and NTS-C2, both NPY+ and TH+ neurons responded to the lower dose of nicotine, whereas the activin and galanin neurons in these regions were unresponsive to either dose of nicotine. In contrast, the higher dose of nicotine was required to activate NPY+ neurons in the A1 region and both NPY+ and galanin+ neurons in the locus coeruleus; the C1 region was unresponsive to nicotine. Since plasma ACTH is elevated by the low dose of nicotine and only NTS neurons are activated by this dose, NPY projections from the NTS are likely to contribute to nicotine stimulated ACTH secretion, in addition to the previously described catecholaminergic neurons. PMID- 9221947 TI - Glutamate-evoked currents in acutely dissociated neurons from the rat medial preoptic nucleus. AB - Membrane currents evoked by glutamate were investigated in acutely dissociated neurons from the medial preoptic nucleus (MPN) of rat. Rapid application of glutamate induced a fast current component in all neurons studied. In addition, in > 50% of the neurons, a slow current component was elicited. The fast and the slow current components were selectively blocked by the AMPA-receptor antagonist NBQX and by the NMDA-receptor channel blocker MK-801, respectively. Rapid application of AMPA induced, in all neurons tested, currents with properties similar to the fast component of the glutamate-evoked currents whereas rapid application of NMDA induced, in approximately 75% of the neurons, currents similar to the slow component of the glutamate-evoked currents. The NMDA-evoked currents showed a marked outward rectification that was attributed to a potential dependent block by extracellular Mg2+. The NMDA-evoked currents also required the presence of extracellular glycine in the micromolar range. In conclusion, the results show that MPN neurons respond to glutamate with currents that can be attributed to activation of ionotropic glutamate receptors of the AMPA-receptor type as well as of the NMDA-receptor type. PMID- 9221948 TI - Heparan sulfate modifies the effects of basic fibroblast growth factor on glial reactivity. AB - Our previous studies have shown that injection of basic fibroblast growth factor (bFGF) into a brain wound enhances astrocyte hypertrophy and macrophage-microglia proliferation in areas adjacent to the lesion. In the present study, designed to test the effects of co-administration of bFGF and heparan sulfate (HS), rats received injections of 200 ng bFGF, 200 ng bFGF with 50 microg HS, or 50 microg HS into a brain wound. Glial proliferation and astrocyte hypertrophy were evaluated in seven non-overlapping subfields in the mid-cortex including the wound edge. Our results show that bFGF-HS, compared to bFGF or HS alone, enhanced the total area of GFAP staining in all subfields except the one nearest to the wound edge. The combination of bFGF and HS did not increase total glial or astrocyte proliferation. We propose that the observed effects resulted from a greater diffusion of bFGF-HS complex into the brain parenchyma, where it bypassed low-affinity binding sites that would otherwise sequester free bFGF. Our results suggest that bFGF-HS complex, compared to bFGF alone, may gain entry into the brain more readily, reach higher concentrations and be more effective as a neurotrophic agent. PMID- 9221949 TI - Expression of the proenkephalin gene in cultured astroglial cells: analysis of cell cycle dependence. AB - Glial progenitors strongly express the proenkephalin (PEnk) gene during their proliferation in the subventricular zone of the neocortex. Also in primary culture, astroglial cells from rat neocortex produce PEnk mRNA. Since the basal expression sharply declines after the cultured cells reach confluence, it seems to be related to cell proliferation. In contrast, activation of protein kinases A and C strongly enhances the levels of PEnk mRNA in confluent cultures. Therefore, it was investigated in cultured neocortical astroglial cells, whether the basal and stimulated expression of the PEnk gene occurred during different phases of the cell cycle. Activation of protein kinases A and C with 8Br.cAMP and tetradecanoylphorbolacetate, respectively, enhanced the PEnk gene expression only during the G1 phase. Unstimulated astroglial cells contained PEnk mRNA during the G1 as well as the S phase of the cell cycle. Since confluent astroglial cells are arrested in the G1 phase, they should have a basal expression of the gene which is comparable to that in preconfluent cells. Thus, the reduced basal expression after confluence indicates that other mechanisms may play a role, such as humoral factors or contact inhibition. PMID- 9221951 TI - Establishment and characteristics of a practical and useful astrocyte cell line transformed by a temperature-sensitive mutant of simian virus 40. AB - A practical mouse astrocyte cell line (A640-IG) was established by transformation with a temperature-sensitive mutant of simian virus 40 (SV40) and the relationship between the function of SV40 large T antigen and the growth and differentiation of A640-IG cells, which are most clearly dependent on temperature that ever established, was reported. A640-IG cells proliferated actively with expression of large T antigen when they were cultured at 33 degrees C. They had a fibroblast-like appearance, and displayed faint immunoreactivity with an antibody against glial fibrillary acidic protein (GFAP). However, when large T antigen expression ceased at 39 degrees C, the cells did not grow actively and differentiated into astrocytes as demonstrated by both their morphological and immunohistochemical characteristics. Differentiation into astrocytes was more obvious when the cells were plated on bacteriological dishes in high density. Western blotting confirmed immunohistochemical observations. A640-IG cells thus showed contrasting behaviour in terms of cell growth and differentiation depending on the temperature. This unique and practical astrocyte cell line is a useful model for investigating the mechanisms of astrocyte growth and differentiation. PMID- 9221950 TI - Nicotinic acetylcholine receptor antagonist effect of fluoxetine in rat hippocampal slices. AB - We compared the effect of mecamylamine and fluoxetine on the hippocampal noradrenaline (NA) release evoked by nicotine in vitro. Nicotine (100 microM) increased the basal release of [3H]NA from rat hippocampal slices. This effect was blocked by the potent nicotinic antagonist mecamylamine in a dose-dependent manner (IC50 = 0.19 microM). The selective serotonin reuptake inhibitor (SSRI) fluoxetine also antagonised the response to nicotine in a dose-dependent manner with a similar strength (IC50 = 0.57 microM). Our data indicate that fluoxetine has nicotinic acetylcholine receptor antagonist effect in the central nervous system. The possible clinical significance of this finding is discussed. PMID- 9221952 TI - Induction of c-fos gene expression by spinal cord transection in Sus scrofa. AB - Functional responses of primary sensory afferents and spinal cord were monitored in swine subjected to a high cervical (C1) spinal transection. Two and a half hours after transection, dorsal root ganglia and cervical and thoracolumbar spinal segments were processed immunocytochemically for the c-fos gene product, Fos and related antigens. In spinal-transected animals, Fos-like immunoreactivity (FLI) was induced in spinal laminae I, V, VII and X and the intermediolateral cell column but not in sensory ganglia as compared to controls: spinal-intact age matched littermates. Spinal laminae expressing FLI harbor sympathetic and somatic interneurons and may aid in maintaining sympathetic outflow. PMID- 9221953 TI - Effects of chronic ibogaine treatment on cerebellar Purkinje cells in the rat. AB - The present investigation assessed the chronic toxicity of ibogaine on cerebellar Purkinje cells in male Fischer 344 rats. A behaviorally active dose of ibogaine (10 mg/kg, i.p.) was administered to a group of six subjects every other day for 60 days while the control group received an equivalent volume of saline (1 ml/kg). Estimates of Purkinje cell number were determined using the optical dissector/fractionator technique. No significant differences in Purkinje cell number were observed between the ibogaine (243764[+/-32766]) and control groups (230813[+/-16670]). PMID- 9221955 TI - Effects of the 5-HT3 receptor agonist 1-(m-chlorophenyl)-biguanide in the rat kindling model of epilepsy. AB - This study assessed the action of the serotonin3 (5-HT3) receptor agonist, 1-(m chlorophenyl)-biguanide (m-CPBG), against both kindled seizures and kindling development from the rat amygdala (AM). The intracerebroventricular (i.c.v.) administration of 40 microg m-CPBG significantly increased the duration of afterdischarge and bilateral forelimb clonus of generalized kindled seizures. In addition, daily i.c.v. treatment with m-CPBG at the same dose prior to each electrical stimulation to the AM significantly facilitated behavioral and electrographic seizure development and reduced the number of stimulations needed to elicit generalized seizures. The present results indicate that m-CPBG increases the duration of fully kindled seizures and facilitates the developmental seizure process, suggesting an excitatory role of 5-HT3 receptors in the kindling model of epilepsy. PMID- 9221954 TI - Enhancing adenosine A1 receptor binding reduces hypoxic-ischemic brain injury in newborn rats. AB - Hypoxia increases brain adenosine concentrations, which provides neuroprotection through activation of central adenosine A1 receptors. This study was carried out to determine whether PD 81,273, which increases adenosine's binding to A1 receptors, would reduce hypoxia-induced brain injury. PD 81,273 (3 mg/kg, i.p.) decreased by about 50% the weight loss of the left cerebral hemisphere caused by hypoxia-ischemia in neonatal rats. Thus, enhancing adenosine's binding to the A1 receptor decreases hypoxic brain damage. PMID- 9221956 TI - Prenatal stress induces a phase advance of circadian corticosterone rhythm in adult rats which is prevented by postnatal stress. AB - Prenatal and postnatal stressors can have different long-term neuroendocrine effects including modifications of stress-induced corticosterone secretion. However, very little is known about the possible long-term effects of prenatal or postnatal stress on the rhythmicity of basal corticosterone secretion in adult offspring. Corticosterone levels were thus determined at six different time points over 24 h in adult rats whose mothers had undergone restraint stress manipulations. The results demonstrate that prenatal stress induces a phase advance in the evening increase of corticosterone levels, and that this change is prevented by postnatal stress. It thus appears that the circadian system governing the HPA axis is modifiable by a prenatal stress, and remains susceptible to compensatory changes during the postnatal period. PMID- 9221957 TI - Expressions of P-selectin- and HSP72-like immunoreactivities in rat brain after transient middle cerebral artery occlusion. AB - The role of an adhesion molecule such as P-selectin may be important in the pathogenesis of stroke. However, temporal, spatial and cellular profiles of the expression of such a protein have not been fully studied. Change of immunoreactive P-selectin was examined in rat brain after transient middle cerebral artery (MCA) occlusion in comparison with that of 72 kDa heat shock protein (HSP72) which is a well known marker of cell injury. Western blot analyses were performed to ensure the selective detection of immunoreactive P selectin and HSP72 proteins with each antibody using brain samples before and after ischemia. Temporal, spatial and cellular changes of immunohistochemical expressions of P-selectin and HSP72 were evaluated with rat brain sections at 2 and 8 h, and 1, 3 and 7 days of reperfusion after 1 h of MCA occlusion (MCAO). Hematoxylin-eosin (HE) staining was performed to evaluate brain cell damage at 3 and 7 days of reperfusion. Western blot showed a single band at molecular weights of 140 and 72 kDa for P-selectin and HSP72, respectively, only after ischemia. No significant band was observed without primary antibody. P-selectin-like immunoreactivity was not normally present in rat brain sections. However, it was expressed mainly in the post-capillary venules of the cerebral cortex and caudate in the MCA territory with a peak at 8 h to 1 day. The expression was diminished by 3 days of reperfusion. An immunoreactive HSP72 was scarcely present in the cerebral cortex and caudate of the sham control brain. However, the protein was induced in neurons of the MCA territory. The HSP72 induction was gradually intensified from 8 h with peaks at 1 day in the cortex and at 3 days in the caudate. The immunoreactivity decreased by 7 days. Histopathological study with HE staining showed no evident cell damage at 3 and 7 days of reperfusion. The present results indicate that temporal, spatial and cellular differences were present in the expressions of immunoreactive P-selectin and HSP72 proteins. P selectin was expressed from an earlier stage of reperfusion in post-capillary venules, and the expression became maximum at the same time both in the cerebral cortex and caudate. In contrast, HSP72 induction began later in neurons and reached maximum at a different time between the cortex and caudate. PMID- 9221958 TI - The Cochrane Brain and Spinal Cord Injury Group. PMID- 9221960 TI - Georg Agricola (1490-1555). PMID- 9221961 TI - Neurological picture. Acute disseminated encephalomyelitis presenting as multiple cystic lesions. PMID- 9221959 TI - X linked adrenoleukodystrophy: clinical presentation, diagnosis, and therapy. AB - X linked adrenoleukodystrophy (X-ALD) is an inherited disorder of peroxisomal metabolism, biochemically characterised by accumulation of saturated very long chain fatty acids. Accumulation of these fatty acids is associated with cerebral demyelination, peripheral nerve abnormalities, and adrenocortical and testicular insufficiency. The lowest estimated birth incidence is one per 100,000. At least six phenotypes can be distinguished, of which the two most frequent are childhood cerebral ALD and adrenomyeloneuropathy. The X-ALD gene has been identified, but thus far no relation between genotype and phenotype has been found. Diagnosis is relatively easy and can be confirmed reliably, and prenatal testing is possible in affected families. Several therapeutic options, some with promising perspectives, are available. Neurologists and other physicians seem not to be familiar with the many facets of X-ALD. In this review, the clinical presentation, the relative frequencies of the different phenotypes, and the diagnostic and therapeutic options are presented. PMID- 9221962 TI - Mitochondrial disease associated with the T8993G mutation of the mitochondrial ATPase 6 gene: a clinical, biochemical, and molecular study in six families. AB - AIM: To contribute to the establishment of a rational clinical, neuroradiological, and molecular approach to neurogenic muscle weakness, ataxia, and retinitis pigmentosa (NARP) and maternally inherited Leigh's syndrome (MILS). METHODS AND RESULTS: The T8993G mutation in the mitochondrial genome was found in several maternal members of six pedigrees, whose clinical status ranged from no symptoms to severe infantile subacute necrotising encephalomyelopathy (Leigh's disease). In one case a MELAS-like syndrome was documented both clinically and neuroradiologically. Relevant genetic features of the series were anticipation of symptoms through subsequent generations, and the presence of several cases in whom the mutation apparently occurred recently or was new. A uniform distribution of the mutation in many tissues was shown in one patient subjected to necropsy. In general, a good correlation was found between clinical severity and mutation heteroplasmy in readily accessible tissues, such as lymphocytes or fibroblasts. By contrast, a consistent reduction of the mitochondrial ATPase activity, to about half of the normal values, was found in most of the clinically affected cases, irrespective of the amount of mutant mitochondrial DNA. CONCLUSIONS: Although the measurement of ATP hydrolysis in cultured fibroblasts was a reliable, and sometimes instrumental, means to identify T8993G positive patients, the relation between the mutation and the oxidative phosphorylation defect is probably very complex, and its understanding requires more complex biochemical analysis. PMID- 9221963 TI - Disappearance of unilateral spatial neglect following a simple instruction. AB - OBJECTIVES: To clarify the reason why patients with left unilateral spatial neglect fail to copy the left side of a daisy like flower, not continuing to draw petals all around. METHODS: A flower was simplified and a figure was made that consisted of a large central circle and small circles surrounding it. Four patients with typical left unilateral spatial neglect performed copying and arrangement tasks to make this figure. In the arrangement task, they were instructed to arrange small circles all around the printed central circle. RESULTS: The patients' identification of the composition seemed flawless. In the copying task, they showed neglect, leaving a space on the left side. They seemed to adhere to their plan to place the same number of small circles as those of the model figure. By contrast, neglect disappeared in the arrangement task. CONCLUSION: Patients with neglect can draw the figure satisfactorily if they use a spatial strategy to arrange small circles all around. This strategy seems to improve motivation for drawing and awareness for the left space. It is considered that in the copying of figures such as a daisy, failure to use a spatial strategy plays an important part in the appearance of left unilateral spatial neglect. PMID- 9221964 TI - A randomised clinical trial comparing interferon-alpha and intravenous immunoglobulin in polyneuropathy associated with monoclonal IgM. The IgM associated Polyneuropathy Study Group. AB - OBJECTIVES: The polyneuropathy associated with a monoclonal IgM directed to the myelin associated glycoprotein (MAG) is a specific entity with a putative causal link between the IgM and the neuropathy. The small benefit offered by alkylating agents or plasma exchanges in these patients justifies the search for alternative treatments. METHODS: A 12 month multicentre, prospective, randomised, open clinical trial was carried out comparing intravenous immunoglobulin (IVIg; 2g/kg and then 1 g/kg every three weeks) and recombinant interferon-alpha (IFN-alpha; 3 MU/m2 subcutaneously three times weekly). The main end point was a clinical neuropathy disability score (CNDS) after six months of treatment. Twenty patients were enrolled; 10 were assigned to IVIg and 10 to IFN-alpha. RESULTS: At six months, one out of 10 patients treated with IVIg had a CNDS improvement of more than 20% whereas eight out of 10 patients treated with IFN-alpha had such an improvement (P=0.005). The mean CNDS worsened by 2.3 (SD 7.6) (8%) in the IVIg group whereas it improved by 7.5 (SD 11.1) (31%) in the IFN-alpha group (P=0.02). This improvement persisted after 12 months and was mainly related to an improvement of the sensory component (P=0.02) whereas the motor component was unchanged (P=0.39). Electrophysiological data did not show improvement of motor nerve conduction velocities whereas sensory nerve conduction velocities improved in the upper limbs. A decrease in the level of the monoclonal IgM was seen in two patients treated with IFN-alpha. At the end of the treatment, antibody activity to MAG was still detected in the serum of all patients. CONCLUSION: IVIg, as used in this study, did not improve patients with polyneuropathy and monoclonal IgM. By contrast, although its mechanism of action remains to be fully elucidated, IFN alpha was effective in eight out of 10 patients at six months. PMID- 9221965 TI - Clozapine versus placebo in Huntington's disease: a double blind randomised comparative study. AB - OBJECTIVES: To establish the effect of the atypical neuroleptic clozapine on chorea, voluntary motor performance, and functional disability in patients with Huntington's disease. METHODS: Thirty three patients with Huntington's disease participated in a double blind randomised trial. A maximum of 150 mg/day clozapine or placebo equivalent was given for a period of 31 days. Assessments were performed in the week before and at the last day of the trial. Chorea was scored using the abnormal involuntary movement scale (AIMS), the chorea score of the unified Huntington's disease rating scale (UHDRS), and judgement of video recordings. Voluntary motor performance was assessed using the UHDRS motor scale. Patients and their partners completed a questionnaire regarding functional disability. Twelve patients already used other neuroleptic medication, which was kept unchanged during the trial period. Results of neuroleptic naive and neuroleptic treated patients were analysed separately. RESULTS: Clozapine tended to reduce chorea in neuroleptic naive patients only (AIMS); improvement seemed more pronounced in patients receiving higher doses of clozapine. Other measures of chorea (UHDRS chorea score, video ratings) showed no improvement. Clozapine had no beneficial effect on chorea in patients already receiving neuroleptic medication. Voluntary motor performance did not improve with clozapine. Neuroleptic naive patients reported aggravation of functional disability, possibly reflecting the frequent occurrence of side effects. Adverse reactions forced trial termination in six patients and dose reduction in another eight, and consisted mainly of drowsiness, fatigue, anticholinergic symptoms, and walking difficulties. CONCLUSIONS: Clozapine has little beneficial effect in patients with Huntington's disease, although individual patients may tolerate doses high enough to reduce chorea. Because adverse reactions are often encountered, clozapine should be used with restraint in this patient group. PMID- 9221966 TI - Familial cavernous malformations in a large French kindred: mapping of the gene to the CCM1 locus on chromosome 7q. AB - OBJECTIVES: To characterise clinically a large French family affected with cerebral cavernomas and to check for linkage of this condition to chromosome 7. METHODS: A family, originating from Normandy and in which five members had undergone surgery for cavernomas, was extended. All members older than 18 were studied clinically and by neuroimaging. Genetic linkage analysis was conducted using 11 polymorphic microsatellite markers located between D7S502 and D7S479. RESULTS: The family included three generations. Among the 25 members investigated, 11 had an abnormal cerebral MRI, eight of them being symptomatic, and 12 were asymptomatic with a normal MRI. The status of the two remaining members could not be established on the basis of clinical and MRI data. The family reported shares some striking features with other previously linked families--namely, a high clinical penetrance and the presence of multiple lesions within most of the affected members. A lod score of 4.04 was obtained with marker D7S657 with no recombinant. Significant lod scores were also obtained with D7S524 (Zmax=3.32 at 0=0.00) and D7S630 (Zmax=3.44 at 0=0.00). These results establish linkage of the condition found in this family to chromosome 7. Haplotype analysis strongly suggests that the gene is telomeric to D7S802 and centromeric to D7S479. CONCLUSIONS: These data confirm linkage of cerebral cavernous malformations to chromosome 7 in a non-Hispanic family. PMID- 9221967 TI - Extracranial and intracranial vertebrobasilar dissections: diagnosis and prognosis. AB - OBJECTIVES: To compare the diagnosis and prognosis of extracranial versus intracranial vertebral artery dissections without intracerebral haemorrhage. METHODS: Twenty two vertebral artery dissections were defined by intra-arterial angiography and classified in two groups: group 1, nine extracranial dissections (seven patients) and group 2, 13 intracranial dissections (nine patients), involving the basilar artery in five cases. Bilateral dissections were found in 38% of the population. Before angiography, all the patients had been investigated by continuous wave Doppler, colour coded Doppler, and transcranial Doppler. Mean follow up was 44 months. RESULTS: The two most important symptoms of both dissections (81% of patients) were unbearable pain preceding stroke and progressive onset of stroke within a few hours. Severe ultrasonic abnormalities were present in 94% of the patients whereas specific ultrasonic signs (segmental dilation with eccentric channel) were rare (19%) in both groups. Major strokes and brainstem strokes represented respectively 67% and 78% in intracranial versus 43% and 29% in extracranial dissections. Severe sequelae (permanent disabling motor or cerebellar deficit) were more often associated with intracranial (44%) than with extracranial dissections (14%). No recurrence of dissection and no cerebral haemorrhage were found under heparin. Significant factors of poor outcome (P< 0.05) were the initial severity of the stroke and the bilateral location of dissections. CONCLUSION: The combination of a pain and a progressive onset of the stroke, corroborated by ultrasonic findings, could have helped to recognise most of these types of dissections. Intracranial dissections have a poorer prognosis than extracranial dissections. PMID- 9221968 TI - Quality of life after epilepsy surgery. AB - OBJECTIVE: To assess the relation between seizure status and quality of life after surgery for drug resistant epilepsy, using a previously validated quality of life model developed for use in epilepsy. METHODS: A retrospective postal survey was made on 94 patients who underwent surgery for epilepsy between 1986 and 1994, and 36 patients who after investigation during the same period were found to be unsuitable for surgery. A health related quality of life model was used containing validated measures of anxiety, depression, self esteem, mastery, impact of epilepsy, affect balance, stigma, overall health status, and overall quality of life, to examine the relation between postoperative seizure status and quality of life. RESULTS: Overall 47.9% of patients were seizure free after surgery. On all measures seizure free patients scored significantly better than either patients deemed unsuitable for surgery or those having more than 10 seizures per year after surgery. Patients having less than 10 seizures per year obtained intermediate scores. There was no difference between the groups unsuitable for surgery or having more than 10 seizures per year postoperatively. Employment rates were significantly different between groups, 80% of seizure free and 53% of patients having less than 10 seizures per year in gainful employment postoperatively, compared with 28% and 27% of patients having greater than 10 seizures per year or those who were unsuitable for surgery. CONCLUSIONS: Within broad categories, postoperative quality of life is clearly related to seizure outcome, but the study emphasises the importance of long term follow up in defining the tangible psychosocial effects of freedom from seizures. PMID- 9221969 TI - Apolipoprotein E epsilon4 allele decreases functional connectivity in Alzheimer's disease as measured by EEG coherence. AB - OBJECTIVES: The epsilon4 allele of apolipoprotein E (APOE) represents a major biological risk factor for late onset Alzheimer's disease. However, it is still not known whether the APOE genotype affects the progression of the disease, assessed by different functional methods. METHODS: The study sample included 41 patients with probable Alzheimer's disease. Subjects had similar severity of disease, age of onset, and duration of illness, and were subcategorised according to their APOE genotypes: 17 with no epsilon4 allele, 14 with one epsilon4 allele, and 10 with two epsilon4 alleles. The control group consisted of 18 healthy subjects comparable with the patients in age and education. Analysed quantitative EEG (qEEG) variables were the ratio of alpha and theta absolute power and EEG coherence in alpha frequency band, representing major cortical association pathways. RESULTS: There was pronounced EEG slowing in all three patient subgroups compared with the controls for the alpha/theta ratio, but there was no significant difference across the patient subgroups. Patients homozygous for the APOE epsilon4 allele had reduced right and left temporoparietal, right temporofrontal, and left occipitoparietal coherence. Patients without and with one epsilon4 allele showed an overlap between the control group and group with two epsilon4 alleles in coherence measures. CONCLUSIONS: APOE epsilon4 does not influence EEG slowing, an index which reflects severity of the disease in patients with Alzheimer's disease, but seems to be associated with selective decreases in functional connectivity as assessed by EEG coherence. This finding might be of clinical importance when considering different pathogenetic mechanisms. PMID- 9221970 TI - Neuropsychological, psychiatric, and cerebral perfusion correlates of leukoaraiosis in Alzheimer's disease. AB - OBJECTIVE: To examine neurological, neuropsychological, psychiatric, and cerebral perfusion correlates of leukoaraiosis in Alzheimer's disease. METHODS: A consecutive series of patients with probable Alzheimer's disease was assessed with a comprehensive neuropsychological battery, a structured psychiatric evaluation, the unified Parkinson's disease rating scale, MRI, and single photon emission computed tomography with technetium 99m hexamethylpropyleneamine oxime (HMPAO) and regional cerebral perfusion measurements. RESULTS: Patients with Alzheimer's disease and leukoaraiosis were significantly more apathetic and had significantly more extrapyramidal signs than patients with Alzheimer's disease without leukoaraiosis. Patients with Alzheimer's disease with leukoaraiosis also had significantly lower bilateral perfusion in the basal ganglia, thalamus, and frontal lobes than patients with Alzheimer's disease without leukoaraiosis. On the other hand, there were no significant differences between groups in age, duration of illness, depression scores, severity of delusions, or deficits on specific neuropsychological tasks. CONCLUSIONS: Leukoaraiosis in Alzheimer's disease may produce significant basal ganglia, and thalamic and frontal lobe dysfunction, which may be associated with more severe apathy and extrapyramidal signs. PMID- 9221971 TI - Abnormal response to negative feedback in unipolar depression: evidence for a diagnosis specific impairment. AB - OBJECTIVES: To assess in further detail the specific form of motivational impairment influencing neuropsychological performance in depression oversensitivity to perceived failure. The present study considers two questions: firstly whether this is specific to depression and secondly how the effect relates to clinical features. METHODS: Unipolar depressed patients and matched controls were assessed on two neuropsychological tests giving explicit performance feedback. The data were analysed in two separate studies to consider the questions above. The first study considered the specificity of the effect to depressed patients, using data on the same tests collected from other patient groups. The second study was a longitudinal assessment of the depressed patients on clinical recovery to determine whether the effect is specific to the depressed state. RESULTS: The effect was not seen in non-depressed patient groups, either neurological or psychiatric groups. The longitudinal study showed a residual abnormal response to negative feedback on clinical recovery. CONCLUSIONS: Abnormal response to negative feedback is specific to a primary diagnosis of depression and may be a trait rather than a state factor of the disorder. These results are discussed in relation to the putative neuropathology of depression and also to cognitive and behavioural accounts of the disorder. The findings presented here have important implications for establishing a link between mood and cognition in unipolar depression. PMID- 9221972 TI - Clinical characteristics of patients with motor disability due to conversion disorder: a prospective control group study. AB - OBJECTIVES: Previous studies have suggested associations between conversion and many different clinical characteristics. This study investigates these findings in a prospective design including a control group. METHODS: Thirty consecutive patients with a recent onset of motor disability due to a conversion disorder were compared with a control group of patients with corresponding motor symptoms due to a definite organic lesion. Both groups had a similar duration of symptoms and a comparable age and sex profile and were assessed on a prospective basis. Background information about previous somatic and psychiatric disease was collected and all patients were assessed by means of a structured clinical interview linked to the diagnostic system DSM III-R, the Hamilton rating depression scale, and a special life events inventory. RESULTS: The conversion group had a higher degree of psychopathology with 33% of the patients fulfilling the criteria for psychiatric syndromes according to DSM-III-R axis I, whereas 50% had axis II personality disorders compared with 10% and 17% respectively in the control group. Conversion patients also had significantly higher scores according to the Hamilton rating depression scale. Although patients with known neurological disease were not included in the conversion group, a concomitant somatic disorder was found in 33% of the patients and 50% complained of benign pain. The educational background in conversion patients was poor with only 13% having dropped out of high school compared with 67% in the control group. Self reported global assessment of functioning according to the axis V on DSM IV was significantly lower in conversion patients, who also registered significantly more negative life events before the onset of symptoms than controls. Logistic regression analysis showed that low education, presence of a personality disorder, and high Hamilton depression score were significantly associated with conversion disorder. CONCLUSION: The importance of several previously reported predisposing and precipitating factors in conversion disorder is confirmed. The results support the notion that conversion should be treated as a symptom rather than a diagnosis and that efforts should be made in diagnosing and treating possible underlying somatic and psychiatric conditions. PMID- 9221973 TI - A self report measure of affective lability. AB - OBJECTIVES: The development and validation of the Center for Neurologic Study Lability Scale (CNS-LS), the first self report measure of affective lability in patients with amyotrophic lateral sclerosis (ALS). METHODS: Potential questionnaire items were identified through interviews with patients and families and expert review. Potential items, as well as measures of affect intensity, affective lability in psychopathology, and depression were administered to 99 patients with ALS for item selection and the examination of factor structure and construct validity. Test-retest reliability was examined using an additional sample of 31 patients with ALS, and criterion related validity was examined by comparing CNS-LS scores with physicians' diagnoses of affective lability in a sample of 77 patients with ALS. RESULTS: A seven item questionnaire emerged, composed of two subscales measuring labile laughter (four items) and labile tearfulness (three items). The CNS-LS showed a pattern of associations with affect intensity, affective lability in psychopathology, and depression consistent with a scale measuring affective lability. The CNS-LS also showed good test-retest reliability and internal consistency, and successfully predicted physicians' diagnoses of affective lability. An auxiliary subscale measuring labile frustration, anger, and impatience also emerged. CONCLUSIONS: The CNS-LS is a short, easily administered, and psychometrically sound measure of affective lability for use with patients with ALS. It has potential applications as both a clinical screening device and a research tool. The need for future research into the relation of depression as well as labile frustration, anger, and impatience to the syndrome of affective lability in neurological disorders is discussed. PMID- 9221974 TI - Limitations of using a cancer registry to identify incident primary intracranial tumours. AB - The completeness and accuracy of registration of primary intracranial tumours in the Scottish Cancer Registry was compared with a detailed incidence study performed over a two year period (1989-90). Of 228 patients with any primary intracranial tumour in the incidence study, 124 (54%) were identified as intracranial tumours in the cancer registry. The registry excluded benign tumours (although this was not consistent) and so the sensitivity of the registry varied with tumour type (84% for neuroepithelial tumours, 22% meningeal, 29% sellar, 0% cranial nerve). Of the 31 malignant tumours not found in the registry on our initial search, nine were found to have been included between 1989-90 but using different International Classification of Diseases-9th revision (ICD-9) codes or postcodes, and seven were found registered after 1990. Eleven per cent of cases (18/170) identified in the cancer registry were excluded from the incidence study: 11 had evidence of an intracranial tumour before 1989 whereas four definitely did not have an intracranial tumour. The cancer registry therefore significantly underestimated the incidence of all primary intracranial tumours, and of malignant intracranial tumours. Incidence studies must use additional methods to identify all primary tumours. Cancer registries should consider registering all primary intracranial tumours and may improve case ascertainment by screening neuroradiology data. PMID- 9221975 TI - Histological surprise: callosal tuberculoma presenting as malignant glioma. AB - A tumour which on CT is clearly intrinsic, irregular, exhibits patchy enhancement with contrast, and invades periventricular tissues, especially the corpus callosum, is very likely to be a glioma and may not be biopsied. A case is presented with these radiological features in which the tumour proved to be a tuberculoma, with complete clinical and radiological resolution after antituberculous chemotherapy. PMID- 9221976 TI - Cerebral blood flow and cerebrovascular response to acetazolamide in patients with chronic alcoholism. AB - Cerebral blood flow and cerebrovascular response to acetazolamide were studied in 12 patients with chronic alcoholism and 12 age matched healthy controls. Blood flows in the cerebral cortex, thalamus, and putamen were significantly lower in the chronic alcoholic group than in the healthy control group. The increase in blood flow caused by acetazolamide did not show any significant difference between the two groups. These findings suggest that the decreased cerebral blood flow in chronic alcoholism is due to decreased cerebral metabolism. PMID- 9221977 TI - No effect of the alpha1-antichymotrypsin A allele in Alzheimer's disease. AB - The apolipoprotein E (ApoE)-epsilon4 allele is associated in a dose dependent manner to an increased risk for Alzheimer's disease. However, the ApoE-epsilon4 allele effect does not account for all patients with Alzheimer's disease, and the existence of other genetic risk factors has been postulated. Kamboh et al reported an association between Alzheimer's disease and the A allele of alpha1 antichymotrypsin (Aact) gene, which was not confirmed in a larger series more recently analysed. The ApoE and Aact genotypes were analysed in 314 patients with Alzheimer's disease and 173 healthy controls, confirming the dose dependent effect of the ApoE-epsilon4 allele. Nevertheless, even using odds ratios adjusted for age and sex, there was no significant effect of the Aact genotype on Alzheimer's disease or on the ApoE-epsilon4 allele associated risk for Alzheimer's disease. PMID- 9221978 TI - The use of self infused intravenous immunoglobulin home therapy in the treatment of acquired chronic demyelinating neuropathies. AB - High dose intravenous immunoglobulin therapy is becoming the established maintenance treatment for several neurological conditions. Therapeutic immunoglobulin is expensive and its administration is time consuming. Efficacy is variable and must be closely monitored. Here, we show that self infused immunoglobulin at home, combined with daily functional strength diaries to predict dose and dosage intervals, is effective, time saving, and convenient for both patients and medical staff. PMID- 9221979 TI - Tremor and other movement disorders after whiplash type injuries. AB - Movement disorders are usually of central origin, but have been reported in association with peripheral trauma. Injuries to the neck of the whiplash type provide a source of both types of injuries. Six cases are reported in which the temporal relation between the injury and the movement disorder make a causal relation likely. This important cause of disability has not previously been appreciated. PMID- 9221980 TI - Evaluation of NeuroPage: a new memory aid. AB - This report describes NeuroPage, a simple and portable paging system, developed in California by the engineer father of a son with head injury working together with a neuropsychologist. Using an ABA single case experimental design, the efficacy of NeuroPage was evaluated with 15 neurologically impaired subjects all of whom had significant everyday memory problems because of organic memory impairment or because of problems with planning and organisation consequent on frontal lobe damage. Data were analysed with an odds ratio test which takes into account different underlying success rates for each target and calculates an average improvement factor. This test showed a significant improvement between the baseline and the treatment phases for each subject (P<0.05). PMID- 9221981 TI - Epidemiological study of primary intracranial tumours in elderly people. AB - The incidence of primary intracranial tumours in a well defined population of persons older than 70 years (elderly) who resided in Kumamoto prefecture was examined. During the period from 1989 to 1995, primary intracranial tumours were diagnosed in 271 elderly people; of these, 155 (57.2%) tumours were confirmed microscopically. In a mean population of 216,000 people over the age of 70 years, this yields an average annual incidence rate of 18.1 cases/100,000 population/year. The incidence was lower in men (15.2/100,000 population) than women (20.3/100,000 population). The age specific incidence/100,000/year was 23.2 for the 70-74 year age group, 18.1 for the 75-79 year age group, 15.1 for the 80 84 year age group, and 7.6 for persons older than 85 years. The most common tumours were meningiomas (50.6%), followed by malignant gliomas (13.3%), pituitary adenomas (12.9%), schwannomas (6.6%), malignant lymphomas (3.7%), and benign astrocytomas (3.7%). PMID- 9221982 TI - Central neurocytoma of the cervical spinal cord. PMID- 9221983 TI - Cenesthetic hallucinations in a patient with Parkinson's disease. PMID- 9221984 TI - Neuroleptic malignant syndrome-like condition in multiple system atrophy. PMID- 9221985 TI - Non-Hodgkin's lymphoma as a new cause of non-thrombotic superior sagittal sinus occlusion. PMID- 9221986 TI - On the disposition of cryopreserved human embryos: an opinion. PMID- 9221987 TI - Frozen preimplantation embryos: parental responsibility versus laboratory liability. PMID- 9221988 TI - Sex chromosomal anomalies in pregnancies conceived through intracytoplasmic sperm injection: a case for genetic counselling. AB - The prevalence of sex chromosomal anomalies (SCA) is higher after treatment with intracytoplasmic sperm injection (ICSI) than in naturally conceived pregnancies. This finding is not only important in the debate about the genetic safety of ICSI, it also has repercussions on the design of appropriate strategies for prenatal and preimplantation diagnosis in ICSI pregnancies. We discuss here in detail the developmental prognosis of individuals carrying a sex chromosomal anomaly. Major malformations do occur in Turner syndrome, but not so in Klinefelter, the triple X and the XYY syndromes. Infertility represents an almost obligate finding in Klinefelter syndrome, but the latest developments in microassisted reproduction may help to overcome this problem. Importantly, mental retardation does not occur more often in individuals with an SCA than in normal controls. Academic achievement, however, may be somewhat reduced compared with peers. Overall, for most children carrying a sex chromosomal anomaly, a major congenital handicap is not to be expected, and the long-term developmental prognosis is fairly good. Therefore, if an SCA is diagnosed prenatally in an ICSI pregnancy, an unbiased and detailed discussion of the developmental perspectives is warranted. The option of continuing such a pregnancy should be given due consideration. PMID- 9221989 TI - The pathophysiology of ovarian hyperstimulation syndrome--views and ideas. AB - Ovarian hyperstimulation syndrome (OHSS) is a serious complication affecting ovulation induction. Its most severe manifestation takes the form of massive ovarian enlargement and multiple cysts, haemoconcentration and third-space accumulation of fluid. The full-blown clinical syndrome may be complicated by renal failure and oliguria, hypovolaemic shock, thromboembolic episodes, adult respiratory distress syndrome (ARDS), and death. Although the pathophysiology of this syndrome has not been completely elucidated, it seems likely that the increased capillary permeability triggered by the release of vasoactive substance secreted by the ovaries under human chorionic gonadotrophin (HCG) stimulation plays a key role in this syndrome. Several factors such as histamine, serotonin, prostaglandins, prolactin, and a variety of other substances have been implicated in this process in the past. At present, factors belonging to the renin angiotensin system, cytokines including the interleukins, tumour necrosis factor alpha, endothelin-1 and vascular endothelial growth factor (VEGF) are thought to be involved in triggering increased vascular permeability after ovulation induction treatment. This manuscript summarizes the current knowledge of the pathophysiology of ovarian hyperstimulation syndrome with emphasis on the correlation of the various factors with the clinical phenomena of this iatrogenic syndrome. PMID- 9221990 TI - Impact of overnight dexamethasone suppression on the adrenal androgen response to an oral glucose tolerance test in women with and without polycystic ovary syndrome. AB - In order to test the hypothesis that adrenocortical overactivity, possibly related to the stress of testing, may impact on the measurement of circulating androgen concentrations during glucose-induced hyperinsulinaemia, we prospectively screened 10 patients with the polycystic ovary syndrome (PCOS) and nine healthy control women with an oral glucose tolerance test (OGTT), before and after the administration of dexamethasone. Blood sampling was performed at 0, 30, 60, 90, and 120 min following the oral ingestion of 75 g of glucose, before and after the administration of 1.0 mg dexamethasone on the evening prior to testing. Total and free testosterone, androstenedione, dehydroepiandrosterone (DHEA), DHEA sulphate (DHEAS), cortisol, glucose and insulin were assessed during the 2 h OGTT. Women with PCOS had increased basal concentrations of free testosterone, total testosterone, androstenedione, and insulin compared to control women. In women with PCOS an acute decline in circulating concentrations of DHEAS occurred during the OGTT. In PCOS women there were no changes in other ovarian or adrenal androgens during OGTT before or following dexamethasone administration. In control women DHEA concentrations declined during the OGTT. Following overnight dexamethasone suppression in control women, circulating concentrations of DHEAS and testosterone also declined. It is concluded that: (i) in PCOS women only the concentration of circulating DHEAS decreased during glucose-induced hyperinsulinaemia and dexamethasone administration did not further alter androgen responses to an OGTT; (ii) it is possible that, in these hyperandrogenic patients, the insulin-related suppression of adrenocortical testosterone and DHEA is negated by their much greater ovarian secretion of these androgens; (iii) in control women DHEA concentrations acutely declined during the OGTT and the administration of dexamethasone resulted in the acute decline of DHEA, DHEAS, and testosterone; (iv) it appears that the stress related to testing impacts on the androgen response to OGTT, at least in healthy women. PMID- 9221991 TI - Menstrual cycle and appetite control: implications for weight regulation. AB - Hormonal fluctuations associated with the menstrual cycle influence appetite control and eating behaviour. Energy intake varies during the reproductive cycle in humans and animals, with a periovulatory nadir and a luteal phase peak. Patterns of macronutrient selection show less consistency but a number of studies report carbohydrate cravings in the premenstrual phase, particularly in women with premenstrual syndrome. The cyclical nature of food cravings are frequently, but not invariably, associated with depression. Fluctuations in appetite, cravings and energy intake during the menstrual cycle may occur in parallel with cyclical rhythms in serotonin, which can be accompanied by affective symptoms. The premenstrual phase can be considered as a time when women are especially vulnerable to overconsumption, food craving and depression; this is often associated with low serotonin activity. PMID- 9221993 TI - Pulsatile gonadotrophin secretion in women with polycystic ovary syndrome after gonadotrophin-releasing hormone agonist treatment. AB - In polycystic ovary syndrome (PCOS), increased luteinizing hormone (LH) pulse frequency has been attributed to either the hypothalamic gonadotrophin-releasing hormone (GnRH) pulse generator or ovarian oestrogen feedback. To address this issue, a detailed examination of pulsatile LH secretion was undertaken during recovery from GnRH agonist (GnRHa) suppression. Each of six women with PCOS and six normal ovulatory women received daily GnRHa treatment for 14 weeks. Frequent blood samples were collected and assayed for gonadotrophins, androgens and oestrogens before, during and up to 4 weeks after treatment. Women with PCOS had higher basal LH pulse frequency and amplitude and increased serum concentrations of LH, androstenedione, testosterone and oestrone than controls. After 3 months of GnRHa treatment, all these parameters were suppressed with no differences observed between the two groups. One week after cessation of GnRHa, LH pulse frequency promptly returned to pre-treatment range with no between-group differences noted, whereas LH pulse amplitude, serum gonadotrophins and ovarian steroids remained maximally suppressed and equivalent in the two groups. Subsequent LH pulse frequency remained constant while LH pulse amplitude and circulating concentrations gradually increased in parallel with a return of serum oestrogen to pre-treatment values. Despite comparable resumption of LH secretion in the two groups, corresponding androgen concentrations in women with PCOS were greater than those of normal ovulatory women. Thus, the immediate restoration of LH pulse frequency after discontinuing GnRHa treatment is largely independent of ovarian oestrogen production and reflects primacy of the GnRH pulse generator in determining basal LH pulse frequency. Equivalent LH pulse frequency rates in the two groups during the recovery period do not suggest an intrinsic hypothalamic pituitary hyperactivity in PCOS. PMID- 9221992 TI - Utility of follicle stimulating hormone (FSH), luteinizing hormone (LH), oestradiol and FSH:LH ratio in predicting reproductive age in normal women. AB - The relative efficacy of follicle stimulating hormone (FSH), luteinizing hormone (LH), FSH:LH ratio and oestradiol is evaluated as a predictor of ovarian reserve (reproductive age) in normal women. Serum levels of FSH, LH, oestradiol and FSH:LH ratios were measured during menstrual cycle days 1-4 in younger (20-25 years; n = 23) and older (40-45 years; n = 32) reproductive age women with regular menstruation and normal reproductive function. On days 1-4, mean levels of FSH, oestradiol and FSH:LH ratios were significantly higher in older compared with younger women. FSH increased in concentration across cycle days in both age groups. A significantly lower LH value in younger versus older women was found only on day 1. Oestradiol showed no change across days in the younger group, but increased significantly from day 1 to day 4 in the older group. FSH values on days 1 or 2 were the best single predictor of age differences. However, the best prediction of age differences was obtained by using the combination of FSH and LH (as opposed to the FSH:LH ratio) on day 1 of the menstrual cycle. PMID- 9221994 TI - Treatment of male infertility due to sperm surface antibodies: IUI or IVF? AB - This prospectively designed study was aimed at comparing the results of two different treatment protocols in 29 infertile couples with proven male immunological infertility, i.e. a positive (>50%) mixed antiglobulin reaction (MAR) test (IgG and/or IgA). In the first protocol (group I, n = 14) couples were treated with ovarian stimulation/ intrauterine insemination (IUI), followed by in vitro fertilization (IVF) if no pregnancy occurred after three IUI cycles. In the second protocol (group II, n = 15), patients were treated with IVF as a first choice procedure. The decision to follow protocol 1 or 2 was made by the couples after information about financial costs and expected success rates (according to the literature) for both treatment options. In group I, nine patients (64.3%) conceived after a maximum of three IUI cycles whereas seven patients (46.6%) of group II became pregnant during the first IVF cycle. The take-home baby rate per started IUI or IVF cycle was 27.3% (9/33) and 44.4% (16/36) respectively with a take-home baby rate of 64.3% after three IUI cycles and 93.3% after three IVF attempts. To conclude, both IUI and IVF yielded unexpectedly high pregnancy rates in this selected group of patients with long-standing infertility due to sperm surface (predominantly IgG) antibodies. Since cost benefit analysis comparing superovulation IUI with IVF may favour a course of four IUI cycles, we advocate superovulation IUI as the first line therapy in male immunological infertility. PMID- 9221995 TI - Effect of antiphospholipid antibodies in women undergoing in-vitro fertilization: role of heparin and aspirin. AB - To describe the prevalence of antiphospholipid antibodies in women undergoing in vitro fertilization (IVF) and to determine if heparin and aspirin affect implantation rates, 191 women with a history of infertility undergoing IVF were prospectively tested for antiphospholipid antibodies. This was a two-centre, non randomized comparison of women with positive antiphospholipid antibodies receiving heparin and aspirin versus standard treatment. An enzyme-linked immunosorbent assay, with referenced standards and known positive and negative sera on each plate, was utilized to measure antibodies to cardiolipin, phosphatidylinositol, phosphatidylglycerol, phosphatidylserine and phosphatidylethanolamine. Statistical analyses of results included analysis of variance and Fisher's two-tailed exact test. Antiphospholipid antibodies were detected in 18.8% of patients undergoing IVF compared with only 5.5% in the 200 normal controls, 26% in 200 women with recurrent pregnancy loss, and 32% in 200 women with systemic lupus erythematosus. In conclusion, antiphospholipid antibodies were found more frequently in women undergoing IVF than in the normal control population. Although implantation rates appeared higher in the group of women treated with heparin and aspirin, no statistically significant differences were detected in implantation, pregnancy and ongoing pregnancy rates between those who received standard therapy and those treated with heparin and aspirin. PMID- 9221996 TI - Transfer of cryopreserved-thawed pre-embryos in a cycle using exogenous steroids without prior gonadotrophin-releasing hormone agonist suppression yields favourable pregnancy results. AB - We have analysed the use of a programmed cycle of administration of exogenous steroids without prior suppression with a gonadotrophin-releasing hormone agonist (GnRHa) for the transfer of cryopreserved-thawed pre-embryos. From July 1992 to June 1994, 199 cycles (162 patients) were studied. Pre-embryos had been previously cryopreserved at the pronuclear stage using 1.5 M 1,2-propanediol as a cryoprotectant. Preparation of the endometrium was achieved in a step-up regime with transdermal oestradiol patches (0.1 to 0.4 mg). Progesterone in oil (50 mg i.m.) was started on cycle day 13. Pre-embryos were thawed on day 14 and transferred on day 15 after evidence of survival and cleavage. The mean (+/- SD) age of patients undergoing transfer was 35.4 +/- 4.3 years. The mean number of pre-embryos thawed was 4.7 +/- 1.8 with a mean of 3.3 +/- 1.4 pre-embryos being transferred. Eight of the cycles demonstrated follicular development >16 mm prior to thaw and transfer; however, these patients did not demonstrate a luteinizing hormone surge. Mean endometrial thickness on day 13 was 10.8 +/- 2.1 mm. Overall pregnancy rate was 29.2% (57/195). The ongoing or delivery rate was 16.1% (32/195). The rate of preclinical losses per transfer was 6.2% (12/195). Overall, the implantation rate was 6.2% (47/757). Thus, the use of a programmed cycle for cryopreserved embryo transfer yields favourable pregnancy outcome and offers practical advantages to patients. Prior suppression with a GnRHa is not necessary for endometrial preparation. PMID- 9221997 TI - Impact of the learning curve on term delivery rates following laparoscopic salpingostomy for infertility associated with distal tubal occlusive disease. AB - The objective of this study was to determine the impact of the learning curve of one surgeon on the term delivery rate following laparoscopic salpingostomy for tubal infertility. This was a retrospective audit of ongoing clinical practice, undertaken in two tertiary level infertility programmes. Subjects in this study were women undergoing surgery for total occlusion of the distal Fallopian tube. The main outcome measure was cumulative term delivery rates. On stepwise life table analysis the length of infertility, primary and secondary infertility, tubal diameter and whether surgery was performed in the first or second half of the series were significantly associated with outcome. These data suggest that there is a learning curve in obtaining skills to perform laparoscopic salpingostomy, that patient selection may improve with experience, and that selection criteria should be emphasized during didactic teaching and the preceptorship process. PMID- 9221998 TI - A simplified quantitative cervical mucus penetration test. AB - A simplified quantitative cervical mucus penetration test was developed and applied to the semen of 21 prospective donors in order to evaluate its performance. Analysis of the data using stepwise multiple regression revealed that the concentration of spermatozoa in mucus was related primarily to the concentration of progressive spermatozoa in the semen (63% of variance explained), and secondarily to the average path velocity of spermatozoa in semen (70% of variance explained by a model incorporating both variables). However, consistent with previous studies, the derived indices of penetration efficiency including the percentage successful collisions (PSC), percentage successful entries (PSE) and the motile density ratio (MDR) were not significantly correlated with any of the semen variables examined, although multiple regression did derive models which explained 27-31% of the variance in these three dependent variables. The strong correlation and correspondence between the PSC and the MDR suggests that the latter may provide a simplified index of penetration efficiency for routine clinical use and further research in this area. PMID- 9221999 TI - Multiple attempts at embryo transfer: does this affect in-vitro fertilization treatment outcome? AB - In this study, we retrospectively analysed data from 877 patients who had 1204 embryo transfer procedures following in-vitro fertilization (IVF) at Midland Fertility Services, UK, between January 1991 and December 1995 to investigate the factors contributing to failure of embryo transfer at first attempt and the impact of immediate retransfer of retained embryos on the treatment outcome. Embryos were significantly more likely to be retained when the embryo transfer catheter was contaminated with mucus (3.3 versus 17.8%, P = 0.000001) or blood (3.3 versus 12%, P = 0.00001) and when the transfer procedure was difficult compared with when it was easy (20.3 versus 0.8%, P = 0.00001). There was no significant difference in the clinical pregnancy rate between those who had all their embryos transferred at the first attempt (24.7%) and those who required more than one attempt (23.2%). The types of embryo transfer catheter used in the unit did not show any difference in terms of embryo retention. Although we recommend aspiration of cervical mucus in order to reduce the rate of retained embryos, there is no evidence from our study to suggest that pregnancy rate is compromised when embryos are retained, provided they are discovered and immediately retransferred into the uterine cavity. Immediate retransfer is more convenient to the patients and reduces the laboratory workload without compromising the treatment outcome. PMID- 9222000 TI - Female infertility: treatment options for complicated cases. The ESHRE Capri Workshop. European Society for Human Reproduction and Embryology. PMID- 9222001 TI - Influence of co-culture with established human endometrial epithelial and stromal cell lines on sperm movement characteristics. AB - The effects of co-culture of human spermatozoa with human immortalized endometrial cells - epithelial or stromal - on sperm movement characteristics, including hyperactivation, were studied using computer-assisted sperm analysis (CASA). Epithelial and stromal cell types could be separated following 8-10 days of culture of endometrial cells originating from human biopsies. Both cell types were immortalized by the SV 40 large T antigen. Co-incubation of sperm with epithelial and stromal monolayers enhanced the rate of hyperactivation: 24.9% (P <0.05) and 17.8% (P = 0.05) versus 9.5% as control, respectively, whereas the majority of motility parameters remained unchanged. Conditioned media had no effect upon sperm parameters, including hyperactivation. Co-incubation with either monolayer was able to maintain sperm motility over a longer period than incubation in control medium alone. In four patients whose spermatozoa did not exhibit hyperactivation, co-incubation with epithelial cells, but not conditioned medium, allowed normal rates of hyperactivation (range: 6.9-15.6%). PMID- 9222002 TI - Intracytoplasmic injection of spermatids retrieved from testicular tissue: influence of testicular pathology, type of selected spermatids and oocyte activation. AB - Spermatid microinjection into oocytes has proven to be a successful assisted reproduction procedure in the animal model and in the human species, since in the latter a few full-term pregnancies were actually obtained. Patients entering our spermatid injection study included those with a total absence of spermatozoa in the testicular tissue notwithstanding previous positive biopsies (n = 29): an obstructive problem (n = 3), secretory azoospermia (n = 26), and those with total arrest at the spermatogenesis level in previous explorative biopsies (n = 15). In the latter group, absence of spermatids was recorded in four cases. Mature, elongated, elongating and round spermatids (ROS) were injected in respectively 3, 2, 3, and 32 attempts. A total of 260 metaphase II oocytes were injected with ROS, 36 oocytes with spermatids at other stages of maturity. The rates of oocytes showing two pronuclei (2PN) and two polar bodies reached 22% and 64% respectively after injection of round or elongated-mature spermatids. The fertilization rate after ROS injection was influenced by the percentage of spermatozoa observed in a previous biopsy. Patients with a positive preliminary biopsy had significantly more 2PN (33%) when compared to those with a severe spermatogenic dysfunction and in whom no spermatozoa were found (only 11%) (P < 0.05). Incubation of oocytes in calcium ionophore after ROS injection had a positive effect on the rate of 2PN formation (36 versus 16%). Ninety per cent of all the normally fertilized oocytes cleaved. The percentage of grade A and B embryos depended on the type of injected cells: 12% after ROS and 30% with the other types of haploid cells. A total of 39 transfers resulted in five pregnancies: three full term with healthy babies delivered (one after ROS injection, and two after injection of an elongating and a mature spermatid), one 4 months ongoing (after elongating spermatid injection) and one miscarriage at 4 weeks (after elongated cell injection). Compared to our conventional intracytoplasmic sperm injection-testicular sperm extraction (ICSI TESE) programme, the implantation rate after ROS injection was very low (5.5 versus 10.5%). PMID- 9222003 TI - Effect of abnormal sperm head morphology on the outcome of intracytoplasmic sperm injection in humans. AB - The present study was designed to determine the efficacy of intracytoplasmic sperm injection (ICSI) using spermatozoa with abnormal head morphology in 17 cases with total teratozoospermia. A total of 160 oocytes were retrieved and 144 metaphase II oocytes were injected. The fertilization and cleavage rates were 50.7 and 93.2% respectively. Fertilization failure occurred in two couples. A total of 54 embryos were transferred and pregnancy rates per initiated and per embryo transfer cycle were 17.6 and 20.0% respectively, while the clinical pregnancy rates per initiated and embryo transfer cycle were 11.8 and 13.3%. The implantation rate was 3.7% (2/54). Out of two pregnancies achieved, one resulted in abortion in the first trimester. The ongoing pregnancy rates per initiated and embryo transfer cycle were 5.88% (1/17) and 6.6% (1/15) respectively. Although the implantation and ongoing pregnancy rates are very low, ICSI seems to be the only treatment modality in cases where teratozoospermia was total with 100% abnormal head morphology. PMID- 9222004 TI - A simple method for recovering the motile spermatozoa from extremely low quality sperm samples. AB - Recovery of motile spermatozoa from extremely low quality samples for use in the intracytoplasmic sperm injection (ICSI) procedure is difficult. To solve this problem we developed a simple method to recover the motile spermatozoa using a 3% polyvinylpyrrolidone (PVP) droplet. After depositing a sperm pellet into this slightly viscous droplet, motile spermatozoa readily swam out to the clear area while immotile spermatozoa dispersed to a lesser extent, so that motile and immotile cells became clearly separated from each other. A total of 36 ICSI cycles using spermatozoa with extremely low quality characteristics were performed. We recovered the motile spermatozoa from all sperm samples from two sources of poor quality spermatozoa. Thirty-one cycles of ICSI with ejaculate resulted in fertilization and pregnancy rates of 54 and 29% respectively. Five cycles of ICSI with frozen-thawed epididymal spermatozoa resulted in fertilization and pregnancy rates of 70 and 60% respectively. The 3% PVP droplet method is very simple and easy to perform, so it may be useful for recovering the motile spermatozoa from extremely low quality sperm samples used for ICSI. PMID- 9222006 TI - Efficient modification of intracytoplasmic sperm injection technique for cases with total lack of sperm movement. AB - A rapid, simple and efficient method for selecting living spermatozoa for intracytoplasmic sperm injection (ICSI) in cases with total lack of sperm movement is described. The selection is based on a characteristic deformation of living spermatozoa exposed to hypo-osmotic conditions during short sequential exposures to modified culture medium and polyvinylpyrrolidone solution; the osmolarity of both of these solutions is reduced by one half by diluting them with an equal amount of water. The application of the sperm viability selection step in six ICSI treatment cycles with total absence of sperm movement resulted in a fertilization rate of 41.9% and the establishment of two ongoing clinical pregnancies. The method described for the selection of living spermatozoa makes it possible to reach acceptable fertilization rates and to obtain ongoing pregnancies by ICSI in cases with total lack of sperm movement. Because of its simplicity, this method can easily be improvised when the total lack of sperm movement is an unexpected finding made on the day of the planned ICSI. PMID- 9222005 TI - Extended sperm preparation: an alternative to testicular sperm extraction in non obstructive azoospermia. AB - Testicular sperm retrieval for the treatment of non-obstructive azoospermia requires the execution of an invasive procedure, with all its possible attending complications and subsequent long-term effects. This study suggests a new non invasive approach for collection of spermatozoa in these patients: the extended sperm preparation (ESP). ESP consists of conducting a thorough microscopic search through many droplets of ejaculate sediment. ESP was performed for 49 patients; in 17 patients (35%), spermatozoa were found and subsequently used in intracytoplasmic sperm injection (ICSI). Of these preparations, five yielded fewer motile spermatozoa than the number of corresponding oocytes available, and in one patient only non-motile spermatozoa were recovered. The remaining 32 ESP negative patients underwent testicular sperm extraction (TESE) from testicular biopsy. Spermatozoa were found in 16 of 32 biopsies (50%) and subsequently used in ICSI. Fertilization and cleavage rates were comparable in both ESP and TESE groups, yielding four clinical pregnancies in each group (27 and 29% respectively). Embryo morphology was defined as excellent in significantly more cases in the ESP group than the TESE group, and implantation rate appeared somewhat higher in the ESP group (16%) than the TESE group (13%). The ESP technique yields results similar to TESE, and can be applied in cases of non obstructive azoospermia as a prerequisite modality enabling us to avoid testicular biopsy in 35% of cases. PMID- 9222007 TI - Induction of spermatogenesis in isolated hypogonadotrophic hypogonadism with gonadotrophins and early intervention with intracytoplasmic sperm injection. AB - Idiopathic hypogonadotrophic hypogonadism (IHH) is a potentially correctable cause of male infertility. However hormonal treatment is usually a slow process and artificial reproductive techniques such as intracytoplasmic sperm injection (ICSI) might be resorted to before full testicular response has been achieved. We report here an unusual variant of IHH of post-pubertal onset in which early intervention with ICSI was attempted. Our patient was 37 years old and presented with male infertility due to azoospermia and undetectable serum gonadotrophin concentrations. He had an apparently normal pubertal development, a testicular volume of 8 ml, normal pituitary-thyroid and pituitary-adrenal function, as well as normal computerized tomographic appearance of the sella region. A combination of human chorionic gonadotrophin (HCG) and menopausal gonadotrophins (HMG) was administered. Spermatozoa were first detected in the semen after 3 months and reached a concentration of approximately 2x10(6)/ml after 9 months. ICSI was attempted at this point; the spermatozoa had good fertilizing ability and three embryos were obtained and replaced. Unfortunately no pregnancy resulted. Treatment with 5000 IU HCG and 150 IU HMG three times per week was continued and sperm counts rose rapidly thereafter to reach 28.3x10(6)/ml after 16 months of injections. His wife conceived naturally during this period and the pregnancy is now in the second trimester. This case illustrates the good prognosis of the rare patient with IHH of post-pubertal onset when treated with gonadotrophins, and suggests that ICSI procedures should be delayed until final testicular maturation is attained. PMID- 9222008 TI - Leukaemia inhibitory factor expression in human follicular fluid and ovarian cells. AB - Leukaemia inhibitory factor (LIF) is a 43 kDa glycoprotein with a remarkable range of biological actions in different tissue systems. LIF improves the rate of fertilization of mouse oocytes in vitro and up-regulates aromatase enzyme. We postulated that LIF may be an important modulator of ovarian function and may also improve embryo quality in humans. Follicular fluid samples from patients undergoing in-vitro fertilization (IVF) and embryo transfer (n = 123), from women undergoing ovarian stimulation (n = 4) and from women undergoing laparoscopy for tubal ligation during their follicular phase (n = 3) were used. Follicular fluid LIF, oestradiol, and progesterone were measured and embryo quality was assessed. Granulosa-lutein cells were cultured for 3 days in Ham's F-12:Dulbecco's modified Eagle's medium (DMEM). Ovarian stromal cells, isolated by enzymatic dispersion of ovarian tissue, were also cultured in the same medium. Following experimental treatments, LIF mRNA and protein concentrations were quantified. The concentration of LIF was 0.8 +/- 0.3 (mean +/- SEM) pg/ml in pre-human chorionic gonadotrophin (HCG) follicular fluid samples and 13.0 +/- 1.1 pg/ml in post-HCG follicular fluid samples (P < 0.05). LIF levels were undetectable in three follicular fluid samples obtained during unstimulated follicular phase. There was a correlation between follicular fluid LIF and follicular fluid oestradiol concentrations (r = 0.36; P = 0.0001) and the number of grade I embryos (r = 0.62; P = 0.01). LIF mRNA and the protein were expressed constitutively but in low amounts in the ovarian stromal cell cultures. The concentrations of LIF mRNA as well as protein were increased by interleukin (IL)-1alpha and tumour necrosis factor alpha (TNF alpha) in a time- and concentration-dependent manner. Purified granulosa-lutein cells expressed low amounts of LIF mRNA and protein which were not significantly increased by IL-1alpha or TNF alpha. Our findings suggest that HCG stimulates the expression of LIF in follicular fluid. Both granulosa-lutein and ovarian stromal cells express the LIF mRNA and produce the protein. Modulation of LIF in these cells may play an important role in the physiology of ovulation and early embryo development. PMID- 9222010 TI - Transvaginal sonographic appearance of functional ovarian cysts. AB - Knowledge of the nature of ovarian lesions is important in order to establish the correct treatment and, especially, to detect ovarian cysts that do not require surgical treatment. With the purpose of demonstrating the utility of transvaginal sonography with colour Doppler of ovarian functional ovarian cysts, 378 ovarian tumours were studied, comparing sonographic diagnosis with pathological findings. Sensitivity and specificity of colour Doppler transvaginal sonography to detect functional ovarian cysts were 84.6 and 99.2% respectively. Positive and negative predictive values were 98 and 93.5% respectively. In our experience, transvaginal sonography with colour Doppler is a useful technique in the diagnosis of ovarian pathology. PMID- 9222009 TI - The relationship between ovarian vascularity and the duration of stimulation in in-vitro fertilization. AB - The role of transvaginal pulsed colour Doppler ultrasound in the assessment of ovarian vascularity was studied in 196 in-vitro fertilization (IVF) cycles. The changes in ovarian blood flow after gonadotrophin-releasing hormone agonist (GnRHa) down-regulation and human menopausal gonadotrophin (HMG) stimulation were determined. The data obtained showed that the ovarian blood flow was significantly improved by oestradiol secretion (P = 0.05) and human chorionic gonadotrophin (HCG) administration (P = 0.003). Folliculogenesis was affected by blood flow supply. The resistance index (RI) value was significantly different (P = 0.05) according to the duration of ovarian stimulation. Patients with a mean RI value >0.56 had a longer stimulation with a significantly lower mean number of oocytes retrieved (P = 0.01) despite the administration of a standard dose of HMG. The RI value is a good indicator of modifications in ovarian vascularization during stimulation. Doppler blood flow measurement could be used to determine the optimal timing for the beginning of HMG administration in patients undergoing ovarian stimulation after down-regulation for IVF treatment. PMID- 9222011 TI - Polarized light microscopy and digital image processing identify a multilaminar structure of the hamster zona pellucida. AB - The zona pellucida (zona) is a glycoprotein coat that envelopes the oocyte and embryo, binds sperm during fertilization and facilitates transfer of the embryo through the Fallopian tube. Before implantation can occur, the blastocyst must hatch from the zona. Several lines of evidence suggest that the zona is multilaminar. We hypothesized that the multilaminar structure of the zona filaments could be imaged non-destructively with the polarized light microscope. A recent modification of the polarized light microscope (pol-scope), which combines innovations in polarization optics with novel image processing software, allows measurement of birefringence at all points of the image. Hamster metaphase II oocytes were placed on glass coverslips which replaced the bottom of culture dishes, imaged under differential interference contrast (DIC) and pol-scope optics, then digitized and processed to measure birefringence magnitude and orientation. The pol-scope revealed the zona to be divided into outer and inner layers separated by a zone of low retardance. This finding is consistent with filaments in the outer layer oriented tangentially and in the inner layer oriented radially. The multilaminar structure of the mammalian zona suggested by differential lectin binding and by scanning electron microscopy could be imaged non-destructively with the pol-scope. Because the pol-scope provides a non destructive method to identify macro-molecular organization of the zona, it may prove useful in developmental studies of hatching and to direct resection of the zona. PMID- 9222013 TI - The optimal equilibration time for mouse embryos frozen by vitrification with trehalose. AB - To determine the best equilibration time in the cryoprotectant before rapid cooling, 8-cell mouse embryos were exposed to a vitrification solution containing ethylene glycol, Ficoll and trehalose in modified phosphate-buffered saline at 5 degrees C for varying periods of time. They were frozen using an ultra rapid freezing method, thawed in a 20 degrees C water bath and cultured for 24 h with 5 bromo-2-deoxyuridine. Embryo development and the number of sister chromatid exchanges, a sensitive indicator of genetic damage, were observed. The results demonstrated that embryo development after freezing and thawing was similar among the groups exposed for periods of 5-40 min. However, the number of sister chromatid exchanges was significantly smaller in the group exposed for 5 min, indicating that this was the safest equilibration time in the vitrification solution. PMID- 9222012 TI - Human Fallopian tube epithelial cell co-culture increases fertilization rates in male factor infertility but not in tubal or unexplained infertility. AB - In order to investigate the effect of human Fallopian tube epithelial cell co culture on fertilization and cleavage rates in tubal, male and unexplained infertility, oocytes collected from 91 patients were randomized to wells containing Fallopian tube epithelial cell monolayers or conventional culture medium, and inseminated with spermatozoa. Fertilization and cleavage were assessed at 18 and 52 h, respectively. Co-culture significantly increased the fertilization rates over the control values in male infertility (41.67 versus 23.43%, P = 0.00005), but not in tubal infertility (69.33 versus 67.93%) or unexplained infertility (65.93 versus 54.36%). Cleavage rates were not different in co-culture and conventional in-vitro fertilization systems in any of the infertility subgroups. The number of blastomeres was significantly higher in the co-culture group on the day of embryo transfer (3.63 +/- 1.12 versus 3.04 +/- 1.26, P < 0.001). Pregnancy rates were similar in all infertility subgroups. There was no significant association between the number of co-cultured embryos transferred and the pregnancy, abortion and multiple pregnancy rates. It was concluded that human Fallopian tube epithelial cell co-culture clearly improves fertilization rates in male infertility but not in tubal or unexplained infertility. Improved fertilization rates in co-culture may be due to positive effect of co-culture on impaired sperm function. PMID- 9222014 TI - Prognostic factors for the success rate of embryo freezing. AB - To find some prognostic factors for the outcome of frozen-thawed cycles, we have retrospectively analysed all frozen pre-embryos that were thawed during 1993 and 1994 at two in-vitro fertilization (IVF) units in Sweden. Supernumerary pre embryos were frozen from 551 oocyte retrievals and these resulted in 660 frozen thawed cycles which lead to 623 thawed embryo transfers. The outcome of these transfers was 137 clinical pregnancies with a pregnancy rate of 22% per frozen thawed embryo transfers. Women <40 years of age had a higher birth rate than those > or =40 years, 19 and 5% respectively (P < 0.01). Transfers with two and three pre-embryos resulted in pregnancy rates of 23 and 27%, respectively, compared with 14% for transfer of one embryo. A pregnancy resulting from the initial embryo transfers had a predictive value for results of the subsequent frozen-thawed cycle. Embryo grade and cleavage stage at the time of freezing was important for the survival of the frozen-thawed pre-embryos. The pregnancy rate was not influenced by the cleavage stage, but a tendency toward a lower pregnancy rate was seen for the embryos with lower grading. To conclude, cryopreservation seems to be beneficial in women <40 years of age, who have supernumerary pre embryos of good quality for freezing and of which at least two can be transferred. PMID- 9222015 TI - Oocyte morphology predicts outcome of intracytoplasmic sperm injection. AB - To examine the influence of cytoplasmic morphology on the success rate of intracytoplasmic sperm injection (ICSI), the morphology of 837 metaphase II oocytes was assessed after cumulus stripping. The main abnormalities detected were excessive granularity, cytoplasmic inclusions such as vacuoles, smooth endoplasmic reticulum clustering and refractile bodies. Microinjection was performed in 538 oocytes with normal cytoplasm, 142 out of 161 with excessive granularity and 112 out of 138 with cytoplasmic inclusions. Very poor oocytes were not injected. No difference was found in fertilization rate. The embryos achieved cleaved normally and a similar number of good quality embryos among the three groups was noted. The outcome of transfer of embryos derived solely from normal oocytes (group A: 72 patients, 183 embryos) was compared with those from oocytes with cytoplasmic abnormalities (group B: 34 patients, 85 embryos). In group A, 17 clinical pregnancies (24% per patient, implantation rate 10%) were established. In group B, only one clinical pregnancy (3% per patient, implantation rate 1%) was established, from the transfer of embryos derived from oocytes with homogeneous granularity of the cytoplasm. No pregnancy resulted following the transfer of embryos from eggs with cytoplasmic inclusions. The difference was statistically significant. The outcome of ICSI is dependent on the quality of the oocytes retrieved. Normal fertilization and early embryo development were achieved in oocytes with abnormal cytoplasm morphology, but the resulting embryos failed to demonstrate the same implantation potential as those derived from oocytes with normal cytoplasm. PMID- 9222016 TI - The effect of the anaesthetic, Propofol, on in-vitro oocyte maturation, fertilization and cleavage in mice. AB - Propofol is a common anaesthetic agent used for oocyte retrieval procedures during in-vitro fertilization (IVF). The effect of Propofol in vitro on mouse oocyte maturation, fertilization and embryo cleavage was studied. In this study, 551 cumulus-free and 222 cumulus-enclosed oocytes from mice stimulated with pregnant mare's serum gonadotrophin (PMSG) were incubated for 30 min in medium containing 0, 100, 1000 or 10,000 ng/ml of Propofol prior to in-vitro maturation. Also, 325 cumulus-enclosed oocytes from mice stimulated to ovulate with PMSG/human chorionic gonadotrophin (HCG) were incubated for 30 min in similar concentrations of Propofol prior to IVF. Maturation, fertilization and cleavage rates were compared. A significant decrease in the in-vitro maturation rate was observed only when the cumulus-free and cumulus-enclosed oocytes were exposed to 10,000 ng/ml Propofol (P < 0.0074 and P < 0.0001 respectively). Fertilization and embryo cleavage rates were not significantly different compared with the controls. These findings give some reassurance with respect to human IVF. However, further studies on the potential effects of Propofol on implantation and pregnancy outcome following IVF are needed. PMID- 9222017 TI - Risk factors for adenomyosis. AB - In order to analyse risk factors for adenomyosis, 707 consecutive women who underwent hysterectomy between January 1993 and June 1994 at the Clinica Luigi Mangiagalli, Milan, Italy, were interviewed before surgery by trained physicians. Information on the presence of adenomyosis was obtained from pathologic records. Out of the 707 women, adenomyosis was identified in 150 subjects (21.2%). Women who smoked tended to be at decreased risk of the condition: in comparison with women who had never smoked, the risk for current smokers was 0.7 (0.3-1.3) and the risk decreased with number of cigarettes smoked per day, the odds ratios being 0.8 and 0.6 respectively in women reporting fewer than 10 and more than 10 cigarette smoked per day (chi2 trend 3.57, P = 0.06). The frequency of adenomyosis was higher in parous women: in comparison with nulliparae, the odds ratio of the disease were 1.8 [95% confidence interval (CI) 0.9-3.4] and 3.1 (95% CI 1.7-5.5) respectively in women reporting one and two or more births (chi2 trend 20.71, P < 0.01). Likewise, women reporting one or more spontaneous abortions had an odds ratio of 1.7 (95% CI 1.1-2.6) for adenomyosis, in comparison with those reporting no spontaneous abortion. PMID- 9222018 TI - Vascular endothelial growth factor in primate endometrium is regulated by oestrogen-receptor and progesterone-receptor ligands in vivo. AB - We investigated hormonal regulation of endometrial angiogenesis in menstruating primates. This study was designed to demonstrate: (i) that cell-specific vascular endothelial growth factor (VEGF) production and expression in monkey endometrium are regulated by steroid receptor ligands; and (ii) mifepristone (RU 486) alters VEGF production even in the absence of a progestin agonist. Endometrial VEGF production was compared by computer-assisted immunohistochemical analysis during induced hypoestrogenism and after oestradiol, progestin, or antiprogestin (mifepristone) treatment. VEGF gene expression was estimated by quantitative reverse-transcriptase polymerase chain reaction (RT-PCR) in endometrial samples from castrate cynomolgus monkeys, from intact monkeys in the luteal phase, and from monkeys treated for 20 days with levonorgestrel (LNG) or mifepristone. VEGF staining intensities in glandular epithelium and VEGF mRNA expression were highest in hypoestrogenic monkeys. Progestin treatment induced intense VEGF staining in the stroma. Gene expression of VEGF-189, but not other isoforms, was higher in progesterone- and progestin (LNG)-exposed endometria compared to mifepristone-exposed endometria or endometria from anovulatory cycles (P < 0.04). Mifepristone abolished VEGF staining in glandular epithelium almost completely. We conclude that VEGF protein and VEGF mRNA expression levels in primate endometrium depend on the steroidal milieu. Anti-angiogenic effects of mifepristone via suppression of VEGF production might represent a mechanism for its quelling effects on endometrium. PMID- 9222019 TI - The effect of various doses of mifepristone on endometrial leukaemia inhibitory factor expression in the midluteal phase--an immunohistochemical study. AB - Leukaemia inhibitory factor (LIF) is a cytokine which plays an obligatory role in mouse blastocyst implantation. In human endometrium, LIF expression is significantly increased in the mid-luteal phase indicating that LIF may also play an important role in the human. We have previously shown that a single dose of 200 mg of mifepristone immediately post-ovulation is an effective contraceptive method, probably due to inhibition of endometrial development and function. The purpose of this study was to investigate the effect of various doses of mifepristone on endometrial LIF expression. A total of 22 fertile, regularly menstruating women were studied during control and treatment cycles. The subjects were divided into four groups: group I received a single dose of 200 mg of mifepristone on cycle day LH + 2 (n = 7). The subjects in groups II and III were treated with either 5 mg (n = 5) or 2.5 mg (n = 5) once a week for 2 months. Group IV subjects received 0.5 mg per day (n = 5) of mifepristone for 3 months. LIF was measured immunohistochemically in endometrial tissue specimens taken on the corresponding day (cycle day LH + 6 to LH + 8) in hormonally-characterized control and treatment cycles. LIF immunostaining was observed in all controls and located to the cytoplasm of the luminal and glandular epithelial cells and stromal cells. In the treatment cycles the staining of luminal epithelium and stroma was similar to controls, while the glandular staining was reduced in all treatment groups. This study reveals that early luteal phase treatment as well as intermittent or daily low dose treatment with mifepristone reduces endometrial glandular LIF expression at the expected time of implantation. The results further support the contraceptive potential of mifepristone in low doses. PMID- 9222020 TI - Laparoscopic extirpation of an aplastic ectopic uterus in a patient with Mayer Rokitansky-Kuster-Hauser syndrome. AB - The Mayer-Rokitansky-Kuster-Hauser (MRKH) syndrome comprises the combined hypoplasia of the vagina and the uterus. In recent years, variable anomalies of the development of the Mullerian duct [as classified by the American Fertility Society (AFS)] have been operated on by the laparoscopic approach. We describe here a laparoscopic extirpation of an aplastic ectopic uterus in a patient found to have a hypoplastic vagina and ectopic uterus attached to the right pelvic wall. The uterus was linked to a normotopic right Fallopian tube and right ovary which were covered with endometriosis and adhesions. While the uterine vessels and the round ligament could be demonstrated on the right side, these structures were missing on the contralateral left side of the uterus. However, a normal tube and ovary were present on the left pelvic wall. Although not explicitly described in the AFS classification, this constellation most likely corresponds to the AFS classification stage Ie. PMID- 9222021 TI - Chemokine and cyclooxygenase-2 expression in human endometrium coincides with leukocyte accumulation. AB - The endometrium contains a resident population of leukocytes, the number and subtype of which vary throughout the menstrual cycle and in early pregnancy. Factors controlling these fluctuations are unknown, but a combination of proliferation in situ and migration from the vasculature has been proposed. Locally acting inflammatory mediators, including specific chemokines and prostaglandins, have been implicated in these processes. Interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) are potent chemoattractants and activators for neutrophils and monocytes respectively. Locally acting prostaglandins modulate vascular permeability, and a synergistic action of prostaglandin E (PGE) with IL-8 has been described. In the present study IL-8, MCP-1 and cyclooxygenase-2 (COX-2), the inducible isoform of prostaglandin synthase, were all localized in the endometrium by immunohistochemistry throughout the menstrual cycle and in early pregnancy. All three inflammatory mediators were localized to the perivascular cells of blood vessels in endometrium and decidua, and additional immunoreactivity for COX-2 was identified in the glandular epithelium. The intensity of immunostaining was reduced in the periovulatory, early and mid-secretory phases and significantly increased premenstrually. These results further support the hypothesis that there is a premenstrual migration of leukocytes into the endometrium mediated by chemokines. PMID- 9222023 TI - Coital rate and pregnancy-induced hypertension. AB - Previous workers have emphasized the possibility that pregnancy-induced hypertension (PIH) is associated with a change of sexual partner. It is noted here that any such association is far weaker than that between PIH and fecundability (the probability of conceiving in a month at risk). This latter association is so strong as to suggest its closeness to a true cause. Of the determinants of fecundability, coital rate seems the most promising to pursue as a possible cause of PIH. PMID- 9222022 TI - Decreased levels of the potent regulator of monocyte/macrophage activation, interleukin-13, in the peritoneal fluid of patients with endometriosis. AB - Endometriosis is characterized by an increase in the number, activation and secretory activity of peritoneal fluid macrophages. Factors regulating the activation of these cells may be important in the pathophysiology of this disease. In this study we measured by enzyme-linked immunosorbent assay the concentrations of the macrophage inhibitory factor interleukin (IL)-13 in the peritoneal fluid of women with and without endometriosis. It was found that women with endometriosis had significantly lower amounts of IL-13 (95 +/- 9.8 pg/ml) in peritoneal fluid, compared with women without endometriosis (115 +/- 30 pg/ml) (P < 0.01). No cycle-specific variation was evident for either group. Another macrophage inhibitory interleukin (IL-10) was also measured, but no differences between women with (16.1 +/- 13.2 pg/ml) or without (10.3 +/- 5.6 pg/ml) endometriosis were seen. The immunolocalization of IL-13 was assessed in eutopic and ectopic endometrium and in isolated peritoneal fluid cells. Glandular epithelial cells and stromal cells in both eutopic and ectopic endometrium were immunopositive for IL-13. No cycle-specific differences in the immunolocalization of IL-13 were seen. In conclusion, the reduced amounts of IL-13 in the peritoneal fluid of women with endometriosis may lead to a lack of suppression of macrophage activation, thereby contributing to the overall pathogenesis of this disease. PMID- 9222024 TI - Deficient syncytiotrophoblast tumour necrosis factor-alpha characterizes failing first trimester pregnancies in a subgroup of recurrent miscarriage patients. AB - Pregnancy failure in mice has been associated with increased placental concentrations of the pro-inflammatory cytokine tumour necrosis factor-alpha (TNF alpha). To investigate the role of uterine TNF-alpha in human first trimester miscarriage, we have collected human decidual and trophoblast tissue from women (i) undergoing surgical termination of pregnancy (n = 27), (ii) undergoing a sporadic miscarriage (n = 20) and (iii) with a history of recurrent pregnancy loss [>3 consecutive pregnancy losses (n = 26)] undergoing a further miscarriage. Formalin fixed tissues were examined for TNF-alpha mRNA (in-situ hybridization) and protein (immunohistochemistry). In decidua from all three groups, TNF-alpha protein and mRNA were co-localized to the decidual stroma, the luminal surface of some maternal vessels and to the glandular epithelium. Chorionic villi from the normal pregnancy and the sporadic miscarriage group exhibited co-localized TNF alpha protein and mRNA in the syncytiotrophoblast and cytotrophoblast. In the recurrent miscarriage group, however, 63.6% of the biopsies showed positive immunostaining in only the cytotrophoblast, compared with 4.0% of women undergoing surgical termination of pregnancy and 0.0% of women with a sporadic failed pregnancy (P < 0.001). TNF-alpha mRNA was also localized exclusively to this layer. This may be a secondary effect caused by a different mechanism of pregnancy loss unique to this subgroup. PMID- 9222025 TI - Marker chromosomes in fetal loss. AB - The clinical significance of marker chromosomes has remained obscure especially when diagnosed prenatally. Some carriers have terminated their pregnancies. Extensive attempts have been made to characterize these chromosomes whose origin is frequently difficult to ascertain. This brief summary presents an overview of the implications for fetal loss. PMID- 9222027 TI - Cumulative pregnancy and live birth rates after gamete intra-Fallopian transfer. AB - To evaluate the efficacy of gamete intra-Fallopian transfer (GIFT) the Kaplan Meier life table method was used to analyse a patient cohort treated with GIFT between 1991 and 1994. In a tertiary referral centre for reproductive medicine, 1628 women with a median age of 33 years and various causes of infertility were included to calculate cumulative pregnancy and live birth rates. Age and cause of infertility were main factor variables and the study was based on a total of 2941 consecutive GIFT cycles, leading to a first clinical pregnancy, and 3052 cycles, leading to a first live birth. The cumulative pregnancy and live birth rates were 49.6 and 38.8% respectively, after three initiated cycles and 64.1 and 52.0% respectively, after five initiated cycles. The multiple pregnancy rate was 22.6%. The implantation rate of 13.1% after GIFT demonstrates that the developing embryo benefits from a period of exposure within the environment of the Fallopian tube. The present results indicate that approximately 50% of couples will have at least one live baby after five initiated GIFT cycles. Advancing age was a major negative prognostic factor for the cumulative live birth rate because of higher cancellation rates, lower implantation rates and higher pregnancy failure rates. PMID- 9222026 TI - Local relaxin biosynthesis in the ovary and uterus through the oestrous cycle and early pregnancy in the female marmoset monkey (Callithrix jacchus). AB - The pattern of peripheral serum concentration for the peptide hormone relaxin in women points to the possibility of an interesting paracrine function in the cycle and early pregnancy. In order to investigate this physiology in detail, it was decided to examine local relaxin biosynthesis in an established primate model for human female reproductive function, the marmoset monkey (Callithrix jacchus). In this initial study relaxin biosynthesis was assessed using a combination of molecular and immunological techniques through the oestrous cycle in the marmoset monkey. The nucleotide sequence of the full-length relaxin gene transcript was cloned from the marmoset ovary and found to be closely homologous to that of the human H2 relaxin. Using gene specific probes derived from this sequence, RNase protection assays, reverse transcription-polymerase chain reaction (RT-PCR) assays and in-situ hybridization, showed relaxin gene expression within the ovary in theca cells and corpora lutea in the oestrous cycle, increasing in early pregnancy. Relaxin gene expression was also identified at a low level in the uterus and placenta, and at a higher level in the prostate in the male marmoset monkey. Using two different relaxin-specific antisera, relaxin-like immunoreactivity was observed in the ovary with a pattern of distribution coincident with that obtained by in-situ hybridization. Immunoreactivity was also found in the non-pregnant uterus, within the endometrial epithelium of the late proliferative phase and increasing within the glands through the secretory phase. Taken together, the pattern of relaxin peptide and mRNA expression show there is the basis for local relaxin physiology within the ovarian follicle and corpus luteum, and within the uterus during the oestrous cycle in this new world monkey. PMID- 9222028 TI - Endocytotic uptake of small unilamellar liposomes by human trophoblast cells in culture. AB - We evaluated the mechanism of uptake of carboxyfluorescein-containing small unilamellar liposomes of different surface charge by trophoblast cells in culture. Carboxyfluorescein-encapsulated neutral liposomes were prepared by using equimolar concentrations of lecithin and cholesterol. Anionic and cationic liposomes were prepared by adding dicetylphosphate and stearylamine. Trophoblast cells from human term placenta were cultured and incubated on the first day at 37 degrees C with liposome-encapsulated carboxyfluorescein or 500 nM of free carboxyfluorescein. The mechanism of uptake was determined by pre-treating the cells with metabolic inhibitors: 2 mM of sodium azide and 25 mM of deoxyglucose for 30 min. The uptake of liposomes was also evaluated both qualitatively under fluorescent microscope and quantitatively by measuring carboxyfluorescein fluorometrically. The uptake of free carboxyfluorescein and cationic liposomes was comparable. The anionic liposomes were taken up by the trophoblast cells more avidly than the neutral (13.2 +/- 1.6 versus 9.5 +/- 1.4%; P <0.01), cationic (2.9 +/- 0.4%; P <0.001) and the free carboxyfluorescein (2.1 +/- 0.9%; P <0.01). When cells were pre-treated with metabolic inhibitors, the uptake of anionic (5.9 +/- 1.8%; P <0.001) and neutral liposomes (4.0 +/- 0.8%; P <0.01) was significantly reduced, whereas uptake of cationic and free carboxyfluorescein remained unaltered. This study indicates that small unilamellar liposomes are internalized by the trophoblast cells in culture by an energy-dependent pathway; most probably by endocytosis. The neutral and anionic liposomes are internalized more avidly than cationic liposomes. PMID- 9222029 TI - Donor insemination: child development and family functioning in lesbian mother families. AB - Findings are presented of a comparative study investigating the family relationships and the emotional and gender development of children raised in lesbian mother families. A total of 30 lesbian mother families with 4-8 year old children created as a result of donor insemination (DI) were compared with 38 heterosexual families with a DI child and with 30 heterosexual families who had a naturally conceived child. A variety of assessment measures, including a standardized interview and questionnaires from the parents and psychological testing of the child were used to collect the data. The quality of the couples' relationships and the quality of the mother-child interaction did not differ between lesbian mother families and either of the heterosexual family groups. The quality of the interaction between the social mother and the child in lesbian families was superior to that between the father and the child in both groups of heterosexual families. Childrens' own perception of their parents was similar in all family types; the social mother in lesbian families was regarded by the child to be as much a 'parent' as the father in both types of heterosexual families. With regard to their emotional/behavioural development, boys and girls raised in lesbian mother families were well adjusted and their gender role development did not differ from that of children raised in heterosexual families. These results indicate that child and family development in lesbian mother families is similar to that of heterosexual families. PMID- 9222031 TI - Axel Munthe Award 1996--Professor Luigi Mastrioanni Jr. PMID- 9222030 TI - Pregnancy following intracytoplasmic sperm injection treatment with dead husband's spermatozoa: ethical and policy considerations. AB - This paper describes the first pregnancy in a childless widow after intracytoplasmic sperm injection (ICSI) treatment with her deceased husband's spermatozoa which had been stored for nearly 3 years before use. Before his death the husband had received treatment for testicular cancer and he had given the appropriate written consent for the future use of his spermatozoa. Of the 10 eggs injected, six resulted in normal embryos. Three embryos were transferred and the remaining three embryos are currently stored for possible future use. The treatment resulted in a continuing singleton pregnancy. The case demonstrated the suitability of ICSI in those difficult cases where the sperm quality is extremely poor. This success is also compared with a widely debated case of another widow who was refused permission to use her deceased husband's spermatozoa. It is concluded that in the case of posthumous use of frozen spermatozoa, the current laws are conveniently applicable in a chronic illness but not so in an acute illness leading to death. In the light of the wide public debate on the issues raised by this legal case, the UK Government has also decided to conduct a review of consent procedures involving the storage and use of genetic material. PMID- 9222032 TI - Intracytoplasmic injection of donor spermatozoa. PMID- 9222033 TI - Progesterone-induced immunosuppression. PMID- 9222034 TI - Acquired carbapenemases. PMID- 9222035 TI - Once-daily dosing of aminoglycosides: review and recommendations for clinical practice. AB - The use of higher-dose, extended interval (i.e., once-daily) aminoglycoside regimens to optimize bacterial killing is justified by a pharmacodynamic principle of aminoglycosides, namely concentration-dependent killing, and by the partial attribution of the toxicity of aminoglycosides to prolonged serum concentrations. Numerous in-vitro and animal studies have supported using once daily aminoglycoside dosing. Clinical studies show at least equal effectiveness and no greater toxicity when compared with traditional regimens. A dose of 5-7 mg/kg of gentamicin, tobramycin, or netilmicin, with at least a 24 h dosing interval should be employed and a similar regimen can be applied to amikacin dosing. As yet, there are some patient populations that have not been adequately studied to determine whether or not once-daily aminoglycoside dosing would be a better choice than traditional dosing regimens. PMID- 9222036 TI - Mechanisms of the bactericidal activity of low amperage electric current (DC). AB - The mechanisms whereby low amperage (10-100 microA) electric current (DC) is bactericidal were investigated with Staphylococcus epidermidis and Staphylococcus aureus. A zone of inhibition test involving the insertion of an anode and cathode into an agar plate inoculated with a lawn of bacteria was used to study the antimicrobial activity of electric current. A zone of inhibition was produced around the cathode when 10 microA (DC) was applied for 16 h. The diameter of the zone was greatly reduced in the presence of catalase. There was no zone around the cathode when the test was carried out under anaerobic conditions. H2O2 was produced at the cathode surface under aerobic conditions but not in the absence of oxygen. A salt-bridge apparatus was used to confirm that H2O2 was produced at the cathode and chlorine at the anode. The antimicrobial activity of low amperage electric current under anaerobic conditions and in the absence of chloride ions against bacteria attached to the surface of a current carrying electrode was also investigated. Antibacterial activity was reduced under anaerobic conditions, which is compatible with the role of H2O2 as a primary bactericidal agent of electricity associated with the cathode. A reduction in chloride ions did not significantly reduce the antibacterial activity suggesting that chlorine plays only a minor role in the bactericidal activity towards organisms attached to anodal electrode surfaces. The localized production of H2O2 and chlorine and the intrinsic activity due to electric current may offer a useful method for eradicating bacteria from catheter surfaces. PMID- 9222037 TI - Characteristics of quinolone-induced small colony variants in Staphylococcus aureus. AB - Exposure of Staphylococcus aureus to 1 x MIC of the quinolone antibiotic pazufloxacin for 24 h, followed by plating on drug-free media, led to the emergence of small colony variants (SCVs) in addition to large colony variants (LCVs). However, following incubation with 0.25 or 4 x MIC of pazufloxacin, only LCVs were obtained. The SCVs were half as susceptible to pazufloxacin or ciprofloxacin as wild-type S. aureus, while the susceptibilities of LCVs were essentially unchanged. The reduced susceptibilities of SCVs did not result from mutations in the quinolone-resistance-determining regions of DNA gyrase and topoisomerase IV, since the sequences of these genes were identical to those of the wild-type. However, the SCVs accumulated pazufloxacin and ciprofloxacin to a lesser degree than did wild-type. Furthermore, their susceptibility to quinolones was almost unaffected by reserpine or verapamil, suggesting that the reduced uptake resulted from decreased permeability, rather than from an active efflux pump. The ability of various quinolones to induce emergence of SCVs in S. aureus, correlated with the presence of carbon-bonded substituents at the C-7 position of a quinoline or naphthyridine nucleus, or with the presence of a benzoxazine nucleus. In conclusion, pazufloxacin-induced SCVs represent a mutant that one might expect to be rapidly eliminated in vivo and, hence, not to survive as a quinolone-resistant pathogen. This finding suggests a novel approach for development of future quinolones. PMID- 9222038 TI - Comparison of disc diffusion and agar dilution methods for antibiotic susceptibility testing of Campylobacter jejuni subsp. jejuni and Campylobacter coli. AB - The correlation between disc diffusion and agar dilution susceptibility testing of five antibiotics was studied against 145 Campylobacter strains: 99 Campylobacter jejuni subsp. jejuni and 46 Campylobacter coli. The percentages of true results and 95% CI for disc diffusion for resistant strains were 100% (93.2 100%) for tetracycline (53 strains tested), 100% (77.2-100%) for ciprofloxacin (13 strains tested), 86.7% (62.1-96.3%) for nalidixic acid (15 strains tested), 100% (56.6-100%) for erythromycin (five strains tested) and 68.8% (44.4-85.8%) for ampicillin (16 strains tested). The percentages of true results and 95% CI were 97.6-100% and 93.2-100% respectively for 89-140 susceptible strains to the five antibiotics tested. There was a 1.4% major error for nalidixic acid, 0.7% very major error for erythromycin, 5.5% and 1.4% minor and major errors respectively for ampicillin. There was complete agreement for ciprofloxacin and tetracycline. Results of ampicillin susceptibility are not expected to be useful in a clinical setting. The nalidixic acid disc is a marker of ciprofloxacin susceptibility as the nalidixic acid-susceptible strains were susceptible to ciprofloxacin while most of the resistant ones were resistant to ciprofloxacin. Overall, our results suggest that disc diffusion is a reliable, easy and inexpensive susceptibility testing method for C. jejuni and C. coli for erythromycin, ciprofloxacin and tetracycline. Until more erythromycin- and ciprofloxacin-resistant strains are tested to confirm the reliability of this test, the resistance to these drugs needs to be confirmed using the Etest or the agar dilution method. PMID- 9222039 TI - Bactericidal index: a new way to assess quinolone bactericidal activity in vitro. AB - A new method of assessing the bactericidal activity of quinolones and other antibacterial agents, known as the bactericidal index or BI, is introduced and discussed. The BI represents total bacterial kill over a drug concentration range limited to levels achievable in vivo. This method allows the bactericidal activity of drugs to be readily compared. Calculation of BI produces a single value per bacterial strain/antibacterial agent combination that represents overall bactericidal activity at clinically relevant concentrations. As an example, the bactericidal activities of quinolones against Enterococcus faecalis are compared. PMID- 9222041 TI - Bacteriostatic and bactericidal activity of levofloxacin against Rickettsia rickettsii, Rickettsia conorii, 'Israeli spotted fever group rickettsia' and Coxiella burnetii. AB - Levofloxacin, the L-isomer of ofloxacin, is approximately twice as active as ofloxacin against most Gram-positive and Gram-negative bacteria, and has improved intracellular pharmacokinetic and pharmacodynamic properties. The present work deals with the in-vitro activity of levofloxacin against the obligate intracellular bacteria Rickettsia rickettsii, Rickettsia conorii, 'Israeli spotted fever group rickettsia' (Israeli SFGR) and Coxiella burnetii. Fluoroquinolones, including ofloxacin, have previously been shown to be bacteriostatic against Rickettsia spp. and C. burnetii in vitro. They are reliable alternatives to tetracycline therapy for Mediterranean spotted fever, scrub typhus and acute Q fever. Levofloxacin was bacteriostatic against R. rickettsii, R. conorii and the Israeli SFGR at concentrations of 0.5-1 mg/L, as determined by both a plaque assay and a dye uptake assay. It was also bacteriostatic against C. burnetii isolates, including the Nine Mile, Priscilla and Q212 strains, at concentrations of 0.5-2 mg/L, as determined using the shell vial assay. Overall, levofloxacin could inhibit rickettsial growth at concentrations equal to or half of those necessary for growth inhibition by ofloxacin. Levofloxacin was not bactericidal against C. burnetii at concentrations up to 4 mg/L. PMID- 9222040 TI - The bactericidal activity of levofloxacin compared with ofloxacin, D-ofloxacin, ciprofloxacin, sparfloxacin and cefotaxime against Streptococcus pneumoniae. AB - The bactericidal activity of levofloxacin was compared with that of four other quinolones and one cephalosporin against four strains of Streptococcus pneumoniae with varying degrees of penicillin G susceptibility. Levofloxacin was found to be the most bactericidal quinolone at its optimum bactericidal concentration, followed by ofloxacin, ciprofloxacin, sparfloxacin and D-ofloxacin, in that order. There was no correlation between quinolone-susceptibility and penicillin susceptibility with any quinolone tested. To allow a comparison between the bactericidal activity of the quinolones and cefotaxime to be made a bactericidal index was used. Against two penicillin-sensitive pneumococci and a penicillin intermediate pneumococcus, cefotaxime was the most potent antibacterial agent tested, followed by levofloxacin. However, against the penicillin-resistant pneumococcus, levofloxacin was considerably more potent than cefotaxime. The remaining quinolones tested were inferior to levofloxacin. Levofloxacin has enhanced activity against pneumococci compared with clinically available quinolones. In addition, levofloxacin may be the future quinolone of choice in the treatment of penicillin-resistant pneumococci. PMID- 9222042 TI - In-vitro activity of D0870, a new triazole antifungal drug, in comparison with fluconazole and itraconazole against Aspergillus fumigatus and Candida krusei. AB - The activity of the new triazole antifungal D0870 was compared with those of itraconazole and fluconazole against Candida krusei and Aspergillus fumigatus, two fungi showing inherent tolerance of fluconazole. The activity of D0870 resembled that of itraconazole against whole cells of C. krusei, but it was less effective against A. fumigatus. However, the effect on sterol biosynthesis, in terms of the sterol type accumulating and IC50 for in-vitro biosynthesis, appeared similar in both species. The superior antifungal effect of D0870 over fluconazole appeared related to better inhibition of ergosterol biosynthesis in A. fumigatus, but in C. krusei this did not account for the entire difference which may result mainly from reduced efflux of drug. PMID- 9222043 TI - Rarity of transferable beta-lactamase production by Klebsiella species. AB - We report a survey of beta-lactamases and their transferability in Klebsiella spp. isolated from blood during 1992-95. beta-Lactamases were characterized by determination of isoelectric point (pI), by hybridization of plasmid DNA preparations with probes for SHV and TEM sequences and by PCR with SHV- or TEM specific primers. There were 80 isolates of Klebsiella pneumoniae and 22 isolates of Klebsiella oxytoca. Most isolates of K. pneumoniae had a chromosomally encoded SHV-1 beta-lactamase (or a closely related enzyme); K. oxytoca also produced chromosomal beta-lactamases, but these were distinct from SHV-1. Plasmid-encoded beta-lactamases were rare in Klebsiella spp., being found in six (7.5%) isolates of K. pneumoniae and in none of the K. oxytoca. beta-Lactamase activities were relatively low (< 100 nmoles nitrocefin hydrolysed per minute per mg of protein) and ampicillin MICs were < or = 128 mg/L for most isolates of both species. However, all isolates of K. pneumoniae with plasmid-encoded beta-lactamases, three other isolates of K. pneumoniae and three isolates of K. oxytoca had high beta-lactamase activities (> 100 nmoles/mg/min) and very high ampicillin MICs (> or = 1024 mg/L). PMID- 9222044 TI - Rifampicin resistance in Neisseria meningitidis: evidence from a study of sibling strains, description of new mutations and notes on population genetics. AB - To provide direct evidence for the mechanism leading to resistance to rifampicin, two Neisseria meningitidis strains from one clonal lineage (so-called sibling strains) were studied; one of these strains was resistant, the other sensitive to rifampicin. The polymerase chain reaction (PCR) was used to amplify fragments of the known rifampicin resistance region on the rpoB gene coding for the beta subunit of DNA-directed RNA polymerase and the amplimers were then sequenced. In addition to the DNA from the sibling strains, DNA from further strains was analysed, including two Spanish, rifampicin-resistant strains, eight further N. meningitidis strains, strains of four further Neisseria spp. and one reference strain. The results demonstrated how quickly and easily N. meningitidis can acquire resistance to rifampicin, and also suggest a clonal population structure within the collection of strains studied. This finding is discussed with respect to recent studies that indicate a more panmictic population structure within particular serogroups of N. meningitidis. PMID- 9222045 TI - Quinolone-resistance mutations in the topoisomerase IV parC gene of Acinetobacter baumannii. AB - Mutations in the parC gene, which encodes a subunit of topoisomerase IV, were determined in 21 epidemiologically unrelated clinical isolates of Acinetobacter baumannii. Our studies highlight the conserved sequences in the quinolone resistance-determining region of the parC gene from A. baumannii and other bacteria. Nine of ten isolates with MICs of ciprofloxacin of > or = 32 mg/L showed a change of Ser80 to Leu and one showed a change of Glu84 to Lys. These results suggest that ParC from A. baumannii is a secondary target for quinolones and that mutations at residues Ser80 and Glu84, when combined with mutations at Ser83 of GyrA, may render A. baumannii highly resistant to quinolones. PMID- 9222046 TI - Immune response to Fusobacterium nucleatum, Prevotella intermedia and other anaerobes in children with acute tonsillitis. AB - The number of aerobic and anaerobic bacteria was determined in the saliva of 20 children with acute group A beta-haemolytic streptococcal (GABHS) pharyngo tonsillitis, and 20 with acute non-GABHS tonsillitis. Antibody titres to four Gram-negative anaerobic bacilli that reside in the oropharynx (Fusobacterium nucleatum, Prevotella intermedia, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans) were determined in these and 20 control patients. An average of 8.8 aerobic and anaerobic isolates per patient saliva specimens were found during the acute tonsillitis stage in both groups, and 6.9 (in GABHS tonsillitis) and 5.6 (in non-GABHS tonsillitis) 5-6 weeks later. There were 10- to 1000-fold more bacteria in the acute stages of the inflammation in both GABHS and non-GABHS groups. These bacteria were Staphylococcus aureus, Haemophilus influenzae, Moraxella catarrhalis, Peptostreptococcus spp., F. nucleatum, Prevotella spp. and Porphyromonas spp. Significantly higher antibodies levels to F. nucleatum and P. intermedia were found in the second serum sample of patients with non-GABHS pharyngo-tonsillitis (P < 0.001) and GABHS tonsillitis (P < 0.05), as compared with their first sample or the levels of antibodies in controls. The increase in the number of several aerobic and anaerobic bacteria during acute tonsillitis and the increase in antibody levels to F. nucleatum and P. intermedia, known oral pathogens, may suggest a possible pathogenic role for these organisms in acute non-GABHS and GABHS tonsillitis. PMID- 9222047 TI - Emergence of resistant variants of HIV in vivo during monotherapy with the proteinase inhibitor saquinavir. AB - We examined the phenotypic and genotypic properties of virus from the peripheral blood mononuclear cells (PBMC) and plasma of eight HIV-1-infected asymptomatic patients before and during monotherapy with the proteinase inhibitor saquinavir. Susceptibility of primary isolates to drug was assessed in PBMC culture by deriving IC50 and IC90 values. The observed increases in IC50 and IC90 after approximately one year of therapy with a dosage of 600 mg tds suggests the presence of virus resistant to saquinavir in vivo. The magnitude of this altered susceptibility ranged from three-fold to in one case 100-fold. In two patients a greater than eight-fold decrease in susceptibility to saquinavir was observed. Sequencing of the proteinase genes in viral RNA obtained from patient plasma and/or PBMC was carried out by PCR in parallel with sensitivity testing. In each case between nine and 12 clones were analysed. In the two patients from whom virus had greater than eight-fold reduction in susceptibility, a point mutation was observed in the viral proteinase (Leu90--> Met/Ile). Further mutations were observed at residues 36, 71 and 84 in these subjects. In a third patient, in whom an eight-fold increase in HIV IC50 of saquinavir was observed, no mutations were detected in the proteinase; sequencing of proteinase cleavage sites in viral gag pol revealed no significant mutations. In no patient was a Gly48-->Val mutation observed, although this has been associated with resistance in vitro. The Leu90- >Met mutation was observed in five subjects, but a greater than eight-fold phenotypic change in antiviral susceptibility was seen in only two of these. Hence, in vivo, the Leu90-->Met but not the Gly48-->Val mutation is necessary, but not sufficient, for phenotypic resistance to saquinavir in HIV. PMID- 9222049 TI - The pharmacokinetics of teicoplanin in infants and children. AB - The pharmacokinetics of teicoplanin were assessed after a single dose and under multidose conditions in 12 infants and children. Study patients ranged in age from 2.4 to 11 years. Each patient received teicoplanin 6 mg/kg body weight given intravenously over 20-30 min, once daily for five consecutive days. Multiple timed blood and urine samples were obtained over the 6 day sampling period and were analysed for teicoplanin by both microbiological assay and HPLC. Three compartment pharmacokinetic analysis was used to describe the drug's disposition characteristics. Peak and 24 h trough serum teicoplanin concentrations averaged 39.3 and 1.8 mg/L after the first dose with little accumulation observed after 5 days of therapy. Teicoplanin disposition was variable; V(d)ss ranged from 0.31 to 0.68 L/kg, t(1/2)gamma from 6.5 to 18.1 h and CI from 29 to 51 mL/h/kg. A substantial amount of the administered drug distributed rapidly to the largest, third compartment, with egress approximately four-fold slower than ingress. The majority of the drug was excreted unchanged in the urine. Teicoplanin administration was well tolerated by all study subjects. Using the teicoplanin pharmacokinetic data derived in our study, a dose of teicoplanin 8 mg/kg body weight administered every 12 h should achieve target serum trough concentrations averaging 11 mg/L in children. Higher doses, e.g. 15 mg teicoplanin/kg administered every 12 h, may be needed for the treatment of deep-seated staphylococcal infections and/or endocarditis. PMID- 9222050 TI - Concentrations of trovafloxacin in bronchial mucosa, epithelial lining fluid, alveolar macrophages and serum after administration of single or multiple oral doses to patients undergoing fibre-optic bronchoscopy. AB - Concentrations of trovafloxacin were measured in serum, alveolar macrophages, epithelial lining fluid and bronchial mucosa following single and multiple oral doses. Concentrations were determined using a microbiological assay method. There were 18 subjects in the single dose and nine subjects in the multiple dose groups. After single dosing, mean concentrations in serum, alveolar macrophages, epithelial lining fluid and bronchial mucosa at 6, 12 and 24 h were as follows: 6 h, 1.41 mg/L, 19.06 mg/L, 3.01 mg/L and 1.52 mg/kg; 12 h, 0.85 mg/L, 16.22 mg/L, 4.8 mg/L and 1.01 mg/kg; 24 h, 0.37 mg/L, 10.23 mg/L, 0.93 mg/L, and no measurable concentration, respectively. After multiple dosing (approximately 6 h post-dose) the corresponding concentrations were 1.47 mg/L, 34.3 mg/L, 10.21 mg/L and 1.67 mg/kg, respectively. These concentrations exceed the MIC90s for the common respiratory pathogens, Haemophilus influenzae 0.06 mg/L, Moraxella catarrhalis 0.008 mg/L and Streptococcus pneumoniae 0.12 mg/L and suggest that trovafloxacin should be efficacious in the treatment of community- and hospital acquired respiratory infections. PMID- 9222048 TI - Trovafloxacin delays the antibiotic-induced inflammatory response in experimental pneumococcal meningitis. AB - This study evaluates the ability of the new fluoroquinolone trovafloxacin to attenuate the inflammatory burst known to occur after initiation of antibiotic treatment in pneumococcal meningitis. After exposure to trovafloxacin or ceftriaxone for 3 h in vitro, Streptococcus pneumoniae was injected intracisternally (i.c.) into rabbits every 3 h over 9 h (n = 6 for each antibiotic). Ceftriaxone-treated S. pneumoniae induced consistently higher CSF leucocyte counts (median 2568/microL versus 543/microL at 6 h; P = 0.03; 4560/microL versus 2207/microL at 18 h; P = 0.03) than trovafloxacin-treated bacteria. Meningitis induced in rabbits by i.c. injection of live S. pneumoniae was treated with equal doses of trovafloxacin or ceftriaxone i.v. (ten per group). The bactericidal rates of both antibacterial agents in CSF were almost identical. In comparison with ceftriaxone, trovafloxacin resulted in lower tumour necrosis factor (TNF) and interleukin 1beta (IL-1beta) CSF levels 2 h after the initiation of treatment (TNF levels, median 26 U/mL versus 141 U/mL; P = 0.02; IL 1beta levels 455 pg/mL versus 1399 pg/mL; P = 0.02). Twelve hours after initiation of therapy, however, TNF and IL-1beta were higher in trovafloxacin treated animals (TNF, 61 U/mL versus 7 U/mL; P = 0.001; IL-1beta, 4320 pg/mL versus 427 pg/mL; P = 0.006). The increase in CSF lactate was less during trovafloxacin therapy than with ceftriaxone (median: 2.0 mmol/L versus 4.0 mmol/L; P = 0.03). In conclusion, S. pneumoniae treated in vitro with trovafloxacin induced less CSF leucocytosis than ceftriaxone-treated S. pneumoniae. After i.c. inoculation of live S. pneumoniae, trovafloxacin therapy delayed, but did not inhibit, the release of the proinflammatory cytokines TNF and IL-1beta, probably by slowing the liberation of bacterial cell wall components into the subarachnoid space. PMID- 9222051 TI - The pharmacokinetics and protein-binding of fusidic acid in patients with severe renal failure requiring either haemodialysis or continuous ambulatory peritoneal dialysis. AB - Fusidic acid is metabolized and excreted by the liver. It is generally assumed that renal impairment has no effect on serum concentrations. However, there are few data on the pharmacokinetics of fusidic acid in patients with chronic renal failure, particularly in those requiring dialysis. Seven patients with chronic renal failure on haemodialysis were given 500 mg sodium fusidate orally every 8 h for the 48 h between dialysis. Seven patients on continuous ambulatory peritoneal dialysis (CAPD) were given the same dosage regimen for seven doses. Fusidic acid concentrations were measured by HPLC. Accumulation was seen, and in 12 of the 14 patients steady-state pharmacokinetics had not been achieved by the third day. In haemodialysis patients, mean (range) Cmax values for the first dose were 13.0 (2.0-25.5) mg/L and for the sixth dose were 40.5 (10.1-69.9) mg/L. Serum concentrations were not reduced by haemodialysis. In CAPD patients mean Cmax values for the first dose were 16.0 (4.8-33.8) mg/L and for the seventh dose were 33.9 (23.4-61.9) mg/L. Fusidic acid concentrations of 1.0-2.3 mg/L were detected in peritoneal dialysis fluid in six of the seven CAPD patients. In both patient groups there was a tendency towards increased T(1/2) with repeated dosing. Protein-binding of fusidic acid in patient serum samples was 87.6-94.6%. PMID- 9222052 TI - Postantibiotic effect of trovafloxacin on Pseudomonas aeruginosa. AB - The postantibiotic effect (PAE) of trovafloxacin (CP 99,219) was investigated with six strains of Pseudomonas aeruginosa at concentrations equivalent to 0.5, 1, 2 and 4 x MIC. Trovafloxacin exhibited a significant PAE with four out of the six strains of P. aeruginosa studied. The PAE values obtained for trovafloxacin with P. aeruginosa ranged from 0.3 h to 2.3 h, increasing with increased exposure time and trovafloxacin concentration. PMID- 9222055 TI - Antiviral activity of 5,6,7-trimethoxyflavone and its potentiation of the antiherpes activity of acyclovir. AB - A naturally occurring flavone, 5,6,7-trimethoxyflavone (TMF), isolated from the plant Callicarpa japonica, was subjected to antiviral assays. The compound exhibited relatively high inhibitory effects on herpes simplex virus type 1 (HSV 1), human cytomegalovirus and poliovirus. The anti-HSV-1 action was not due to the inhibition of virus adsorption, entry and viral protein synthesis, but might involve, at least in part, a virucidal activity, which results in a suppression of viral binding to host cells at an early replication stage. TMF and acylovir were synergic in their anti-HSV activities at levels below the 50% inhibitory concentrations for antiviral activity. PMID- 9222053 TI - Lack of resistance to erythromycin, rifampicin and ciprofloxacin in 98 clinical isolates of Legionella pneumophila. AB - Ninety-eight consecutive clinical isolates of Legionella pneumophila were tested for erythromycin, rifampicin and ciprofloxacin susceptibility. MICs, determined by agar dilution testing, were in the range 0.06-1 mg/L of erythromycin, 0.007 0.015 mg/L of rifampicin and 0.015-0.03 mg/L of ciprofloxacin. No resistance against the antibiotics tested was detected. It is thus likely that therapeutic failures in legionnaires' disease are not related to the emergence of resistance against commonly used antimicrobial agents. PMID- 9222054 TI - Factors affecting development of rifampicin resistance in biofilm-producing Staphylococcus epidermidis. AB - Selection and regrowth of variants resistant to 0.016-32 mg/L of rifampicin, which were present at a frequency of 10(-7) in the initial inoculum, were seen when large inocula (> 10(5) cfu/mL) of Staphylococcus epidermidis ATCC 35984 were incubated with the drug. Conventional MIC determinations using approximately 10(5) cfu/mL did not detect the resistant variants. Larger inocula increased the MIC by > 8000-fold. Population analysis showed that rifampicin concentrations above the MIC (measured at an inoculum of approximately 10(5) cfu/mL) select highly resistant variants (MIC > 256 mg/L) when large inocula (> or = 10(5) cfu/mL) were incubated with rifampicin. The resistant variants were stable through ten passages. It was not possible to prevent regrowth of the resistant variants by increasing the rifampicin concentration further. At subinhibitory concentrations there was no development of rifampicin resistance. PMID- 9222056 TI - Therapeutic effects of omoconazole nitrate on guinea-pigs experimentally infected with Trichophyton mentagrophytes. AB - The therapeutic effects of topical omoconazole nitrate (OMZ) on Trichophyton mentagrophytes-infected guinea-pigs were investigated. Once-daily topical application of 0.25, 0.5, 1 or 2% OMZ cream preparation for 14 consecutive days, starting on the fifth day after infection, was therapeutically effective. The effectiveness appeared to increase with the concentration of the active preparation, and was nearly maximal at 1%. OMZ cream preparation (1% or 2%) was superior to 1% bifonazole cream preparation in improving local lesions and culture results. This result suggests that topical OMZ may be clinically useful in the treatment of patients with dermatophytosis and probably those with other superficial mycoses. PMID- 9222057 TI - The pharmacokinetics of once-daily oral 400 mg ofloxacin in patients with peritonitis complicating continuous ambulatory peritoneal dialysis. AB - Seven patients with end-stage renal disease requiring support by continuous ambulatory peritoneal dialysis received once-daily 400 mg oral ofloxacin for 7 days for the treatment of bacterial peritonitis. Serum and peritoneal dialysis fluid (PDF) were collected for assay throughout the course of the study and for 5 days thereafter. Ofloxacin, desmethyl ofloxacin and ofloxacin-N-oxide accumulated over the course of therapy and could still be detected in serum and PDF 5 days after the end of therapy. The mean elimination half-life of ofloxacin in serum was 32 +/- 7 h, desmethyl ofloxacin 45 +/- 26 h and for ofloxacin-N-oxide 44 +/- 15 h. The total mean recovery of ofloxacin and its metabolites from the PDF was 15.4%. This regimen results in serum and PDF concentrations likely to be effective for the treatment of infection for at least 10 days. PMID- 9222058 TI - Evaluation of the resistance induction in enteric flora in children caused by oral ampicillin plus sulbactam. AB - To evaluate the effect on bacterial resistance of a beta-lactamase inhibitor, resistance patterns of predominant bacteria in enteric flora were evaluated before and after a 7-day course of oral ampicillin (100 mg/kg/days, qid, in 16 patients) and ampicillin-sulbactam (50 mg/kg/day of ampicillin, bd, in 32 patients) therapy. Ampicillin and ampicillin-sulbactam MICs for Escherichia coli, the predominant bacteria in all cases, and resistance rates of E. coli species to both antibiotics were 51.20 +/- 13.80 mg/L, 87.5% and 4.84 +/- 2.11 mg/L, 21% before the treatment respectively. Post-treatment MICs and resistance rates were 106.51 +/- 14.05 mg/L, 100% and 15.89 +/- 5.76 mg/L, 37.5% respectively, indicating a significant increase in MICs of both antibiotics (P < 0.05), being more prominent in the case of ampicillin-sulbactam (about four-fold). We concluded that oral ampicillin-sulbactam could also decrease the susceptibility of the enteric flora to ampicillin. PMID- 9222059 TI - Evolution of extended spectrum cephalosporin resistance in the pneumococcus. PMID- 9222060 TI - The effect of zinc on imipenem. PMID- 9222061 TI - The region of the parE gene, homologous to the quinolone-resistant determining region of the gyrB gene, is not linked with the acquisition of quinolone resistance in Escherichia coli clinical isolates. PMID- 9222062 TI - Development of in-vivo resistance after quinolone treatment of gonococcal urethritis. PMID- 9222063 TI - Serum bactericidal activity of ceftazidime administered as continuous infusion of 3 g over 24 h versus intermittent bolus infusion of 2 g against Pseudomonas aeruginosa in healthy volunteers. PMID- 9222064 TI - The chemistry and biological profile of trovafloxacin. AB - The fluoroquinolone antibacterials are noted for their activity after oral administration and potent activity against Gram-negative pathogens. Trovafloxacin (CP-99,219) is a new quinolone antibacterial characterized by a novel 3 azabicyclo[3.1.0]hexyl substituent at the C-7 position, which was discovered in the course of a programme targeting improved activity compared with ciprofloxacin against Gram-positive aerobic organisms and anaerobes, as well as an extended elimination half-life. An overview of the chemical properties of trovafloxacin is given. Trovafloxacin exhibits excellent potency against Gram-positive organisms and anaerobes, while retaining the potent Gram-negative activity of ciprofloxacin. Its pharmacokinetic properties in humans have been shown to be compatible with a once-daily dosing regimen. The combined spectrum and pharmacokinetics of trovafloxacin have been demonstrated to result in excellent efficacy in both animal models of infections and human clinical trials. Phase II and Phase III programmes have been completed. PMID- 9222065 TI - In-vitro activity of trovafloxacin against clinical bacterial isolates from patients with cancer. AB - The antibacterial activity of trovafloxacin was compared with that of ciprofloxacin, levofloxacin, ofloxacin, sparfloxacin and norfloxacin against bacterial isolates from patients with cancer. In general, the activity of trovafloxacin was comparable to that of ciprofloxacin, levofloxacin and sparfloxacin against most Gram-negative isolates tested (minor differences in the activity of each agent against individual species were seen) and it was the most active agent tested against Stenotrophomonas maltophilia, inhibiting 80% of these isolates at <2.0 mg/L. Trovafloxacin was also the most active agent tested against Gram-positive organisms, including ciprofloxacin-susceptible strains and most ciprofloxacin- and methicillin-resistant staphylococci and enterococci. It was much more active than ciprofloxacin against streptococci, including Streptococcus pneumoniae and the viridans streptococci, and was also active against Bacillus cereus and Listeria monocytogenes, inhibiting all isolates at a concentration of <0.5 mg/L. PMID- 9222066 TI - Time-kill study of the activity of trovafloxacin compared with ciprofloxacin, sparfloxacin, metronidazole, cefoxitin, piperacillin and piperacillin/tazobactam against six anaerobes. AB - A time-kill method was developed to examine the killing kinetics of trovafloxacin, ciprofloxacin, sparfloxacin, metronidazole, cefoxitin, piperacillin and piperacillin/tazobactam against one strain each of Bacteroides fragilis, Bacteroides thetaiotaomicron, Prevotella melaninogenica, Fusobacterium mortiferum, Peptostreptococcus magnus and Clostridium perfringens. Solutions and suspensions were prepared inside an anaerobic glove box, using syringes and prereduced broth. Bottles were then incubated outside the chamber and viability counts determined after incubation for 0, 6, 24 and 48 h in a shaking water bath, avoiding introduction of air. Bacteriostatic/bactericidal concentrations (mg/L) after 48 h for the six strains were: trovafloxacin, 0.03-1/0.03-1; ciprofloxacin, 0.25-16/0.25-32; sparfloxacin, 0.06-2/0.06-8; metronidazole 1-64/1-64; cefoxitin, 0.125-16/0.125-32; piperacillin, 0.125-64/0.125-64; piperacillin/tazobactam, 0.06 2/0.125-8. Bacteriostatic levels were within two dilutions of broth MICs. By this time-kill method, trovafloxacin had the lowest bacteriostatic concentrations of all compounds tested. PMID- 9222067 TI - Antibacterial activity of trovafloxacin against nosocomial Gram-positive and Gram negative isolates. AB - A total of 1132 clinical isolates from 952 patients with nosocomial infections were tested against the fluoroquinolone trovafloxacin by the agar dilution method. They comprised 285 staphylococci, 111 streptococci, 94 enterococci and 470 isolates of Enterobacteriaceae, 92 Pseudomonas aeruginosa, 27 Stenotrophomonas maltophilia, 28 Haemophilus influenzae and 25 Acinetobacter calcoaceticus. The in-vitro activity of trovafloxacin was compared with that of ciprofloxacin, norfloxacin, beta-lactam and aminoglycoside agents. Over 96% of Enterobacteriaceae were susceptible to trovafloxacin with an MIC of <0.03-4 mg/L. It also inhibited 97% and 100% clinical isolates of P. aeruginosa and S. maltophilia, respectively. All staphylococci, including 51 strains of methicillin resistant Staphylococcus aureus, were susceptible to trovafloxacin, which also had excellent activity against streptococci and enterococci, inhibiting all 111 strains of the former and 94% of the latter. Trovafloxacin had a greater activity against both Gram-positive and Gram-negative bacteria than ciprofloxacin, norfloxacin, penicillins, cephalosporins and the aminoglycosides tested. PMID- 9222069 TI - In-vitro activity of trovafloxacin, a new fluoroquinolone, against recent clinical isolates. AB - Trovafloxacin (CP-99,219) was very active against Gram-negative species examined including Haemophilus influenzae, Moraxella catarrhalis, Legionella spp., Neisseria spp. and Escherichia coli (MIC90s < or = 0.03 mg/L). In general trovafloxacin was twice as active as ofloxacin but only half as active as ciprofloxacin against Gram-negative species. Trovafloxacin was active against Gram-positive organisms, including Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes and Enterococcus faecalis (MIC90s < or = 0.25 mg/L). Against these organisms activity was eight to 16 times greater for trovafloxacin than for either ofloxacin or ciprofloxacin. In addition, Chlamydia spp., Mycoplasma spp. and Ureaplasma urealyticum were eight to 16 times more susceptible to trovafloxacin than to either ofloxacin or ciprofloxacin. These in vitro data show that trovafloxacin is a broad-spectrum fluoroquinolone with greater activity against clinically important Gram-positive species compared with ofloxacin or ciprofloxacin. PMID- 9222068 TI - In-vitro activity of trovafloxacin against sensitive and resistant aerobic bacteria using the standard microdilution broth method and Etest. AB - A comparison of MICs of trovafloxacin (CP-99,219) determined by the standard microdilution broth method versus the Etest was performed for multiple strains of Gram-positive and Gram-negative bacteria. A comparison was also made of the in vitro activity of trovafloxacin versus ciprofloxacin and ofloxacin. The MIC50 and MIC90 were determined by both methods for each species tested. The Etest resulted in MICs one to two dilutions higher than the microdilution broth method. Trovafloxacin was the most active agent against Gram-positive organisms. Ciprofloxacin was the most active agent against Citrobacter freundii, Proteus mirabilis, Proteus vulgaris, Morganella morganii and Serratia marcescens, while trovafloxacin had equal or greater activity compared with ciprofloxacin and ofloxacin against the other Gram-negative organisms tested. Overall, ofloxacin was the least active agent tested. In addition, the in-vitro activity of trovafloxacin or ciprofloxacin in combination with ampicillin/sulbactam, gentamicin or vancomycin was evaluated. The combination of trovafloxacin and gentamicin was synergic against two of 20 Enterococcus faecium isolates, the combination of trovafloxacin and ampicillin/sulbactam was synergic against two of 24 Enterococcus faecalis isolates, and the combination of ciprofloxacin and gentamicin was synergic against one of 25 Stenotrophomonas maltophilia isolates. All other antibiotic combinations resulted in an additive or indifferent effect. PMID- 9222070 TI - In-vitro and in-vivo activity of trovafloxacin against Streptococcus pneumoniae. AB - Trovafloxacin had greater in-vitro activity than comparative fluoroquinolone agents against penicillin-sensitive pneumococci in studies from the USA, UK, Slovakia, Czech Republic, Sweden and South Africa. This activity was maintained against penicillin-resistant strains, with MIC90 values of < or = 0.25 mg/L observed for both groups. Bactericidal activity appeared to occur within one or two dilutions of the MIC and, in the limited number of strains studied, the MIC was independent of the medium tested and pH over the range pH 5-8. Mutation to decreased susceptibility to trovafloxacin occurred in vitro at a low frequency in the pneumococcus (< or = 8.9 x 10(-9)). Mutants with changes in the topoisomerase IV A subunit (GrlA) were still inhibited by 0.5 mg/L of trovafloxacin. Trovafloxacin was more efficacious than ciprofloxacin, temafloxacin or ofloxacin in mouse pneumonia models for both penicillin-susceptible and penicillin resistant pneumococci. Trovafloxacin was also highly efficacious in a rabbit pneumococcal meningitis model. These data suggest that the clinical efficacy of trovafloxacin against pneumococci should be evaluated further. PMID- 9222072 TI - Susceptibility of Chlamydia trachomatis to trovafloxacin. AB - Chlamydia trachomatis infections are a major cause of morbidity in the sexually active. While current therapy is usually effective, isolates demonstrating relative resistance to erythromycin or heterotypic resistance to erythromycin, tetracycline and their congeners have been described, establishing a need to continue to evaluate other antimicrobial agents for possible efficacy. In this study trovafloxacin, a compound related to the fluoroquinolones, was tested in tissue culture for in-vitro efficacy against 19 strains of C. trachomatis, including three strains known to exhibit heterotypic resistance. All strains were fully sensitive to trovafloxacin with a minimum inhibitory concentration at which 90% of inclusions were reduced (IR90) of 0.05 +/- 0.07 mg/L (mean +/- S.D.). The IR90 for the comparison antibiotics erythromycin, doxycycline and ofloxacin were 0.17 +/- 0.07, 0.10 +/- 0.03 and 0.35 +/- 0.15 mg/L respectively. Trovafloxacin is very active on a weight basis and deserves further evaluation. PMID- 9222071 TI - The in-vitro activity of trovafloxacin, a new fluoroquinolone, against Gram positive bacteria. AB - The in-vitro activity of trovafloxacin, a new quinolone, was compared with that of ciprofloxacin, erythromycin, various beta-lactam antibiotics and, where appropriate, clindamycin and vancomycin against a range of Gram-positive bacteria including staphylococci (n = 201), Streptococcus pneumoniae (n = 83), beta haemolytic streptococci (n = 46), viridans group streptococci (n = 100), Streptococcus milleri (n = 18) and enterococci (n = 161) by an agar dilution technique. In addition, time-kill studies were performed to estimate the bactericidal activity of trovafloxacin against S. milleri and viridans group streptococci. Trovafloxacin was the most active agent tested against staphylococci. It also showed good activity, at least four-fold and usually eight to 16-fold that of ciprofloxacin, against all the streptococci. Trovafloxacin showed good activity against vancomycin-sensitive Enterococcus faecalis and Enterococcus faecium, but was less active against the 11 isolates of vancomycin resistant enterococci. Trovafloxacin showed comparable or superior bactericidal activity to amoxycillin against the S. milleri and viridans group streptococci tested. PMID- 9222073 TI - A study of the phototoxic potential of trovafloxacin in BALB/c mice. AB - Trovafloxacin, a broad-spectrum naphthyridone antimicrobial agent, was evaluated for potential phototoxicity in a standardized in-vivo test system that has been used previously to assess quinolone antibiotics. Fasted BALB/c mice were given a single oral dose of either trovafloxacin mesylate (10, 30, 90 or 250 mg/kg) or the positive control lomefloxacin hydrochloride (71 mg/kg) and immediately exposed to long-wave ultraviolet (UVA) light. Animals were irradiated for 4 h, equal to a total UV light irradiation of approximately 18 J/cm2. Before dosing, at the end of the irradiation period and at approximately 24, 48, 72 and 96 h after dosing, both ears of each mouse were evaluated for changes indicative of a positive response: erythema, oedema or a measurable increase in ear thickness. Under the conditions of this study, trovafloxacin produced a mild response (erythema and a slight increase in ear thickness) in mice given a dose of 90 or 250 mg/kg; no significant response was observed in mice given either lower doses (10 or 30 mg/kg) or the vehicle. In contrast, 71 mg/kg of lomefloxacin produced a strong and persistent phototoxic response. The results of this study demonstrate that the phototoxic potential of trovafloxacin is considerably less than that of lomefloxacin and, when compared with similar studies with related compounds, suggest that trovafloxacin is among the least phototoxic of the fluoroquinolone class. PMID- 9222074 TI - Pharmacokinetics and safety of trovafloxacin in healthy male volunteers following administration of single intravenous doses of the prodrug, alatrofloxacin. AB - Fifteen healthy male volunteers (in four groups) received single 1 h i.v. infusions of alatrofloxacin (CP-116,517) equivalent to 30, 100, 200 or 300 mg of its active metabolite, trovafloxacin (CP-99,219). Blood and urine were sampled over 73 and 72 h, respectively, and plasma levels of alatrofloxacin and serum concentrations of trovafloxacin were determined by HPLC with UV detection. Alatrofloxacin was not detectable in plasma samples collected after the end of infusion, indicating rapid conversion to trovafloxacin. Maximum serum concentrations of trovafloxacin were achieved at the end of the infusions. Mean maximum plasma trovafloxacin concentrations for the four alatrofloxacin doses were 0.4, 1.8, 2.3 and 4.3 mg/L. The mean area under the concentration-time curve increased proportionally with the dose. The elimination half-life (T(1/2)) for trovafloxacin was independent of the dose and the mean T(1/2)s for the 100, 200 and 300 mg equivalent doses of alatrofloxacin were 10.4, 12.3 and 10.8 h. Approximately 10% of the equivalent dose was recovered as unchanged trovafloxacin in the urine. No clinical adverse or laboratory reactions were associated with i.v. administration of alatrofloxacin and its conversion to trovafloxacin. These results indicate that alatrofloxacin is rapidly converted to trovafloxacin and that the pharmacokinetic parameters for this new fluoroquinolone after i.v. administration of its parent compound are similar to those reported after oral administration of equivalent trovafloxacin doses. PMID- 9222075 TI - Effect of trovafloxacin, a new fluoroquinolone antibiotic, on the steady-state pharmacokinetics of theophylline in healthy volunteers. AB - Some fluoroquinolone antibiotics interfere with theophylline clearance, thereby raising concentrations of circulating theophylline and increasing the potential for toxicity. The effect of steady-state serum concentrations of the new fluoroquinolone trovafloxacin on the steady-state pharmacokinetics of theophylline was examined in 12 healthy male volunteers. For 7 days, the subjects received morning and evening theophylline doses adjusted to achieve steady-state plasma concentrations of 8-15 mg/L, the lower end of the therapeutic range. From day 8 to day 15, six volunteers received, in addition to theophylline, 200 mg of trovafloxacin in the morning and placebo in the evening (group A) and six received placebo twice daily (group B). Serial plasma samples obtained over 12 h and 60 h after the morning theophylline dose on days 7 and 14, respectively, were analysed for theophylline by HPLC with UV detection. There were no significant differences in mean Cmax or AUC(0-12) between the two groups on day 7 or on day 14, nor were there significant within-group differences on the two days. On day 14, mean Cmax, AUC(0-12) and T(1/2) (measured on day 14 only) in group A were 10.15 mg/L, 107.32 mg x h/L and 9.0 h, respectively. In group B, the values were 10.81 mg/L, 113.73 mg x h/L and 8.3 h, respectively. The study drugs were well tolerated, and no clinically significant changes in vital signs or laboratory test values were noted. We conclude that steady-state concentrations of trovafloxacin have no clinically significant effect on the steady-state concentrations of theophylline within the therapeutic range in healthy subjects. PMID- 9222076 TI - Oral bioavailability of trovafloxacin with and without food in healthy volunteers. AB - Two studies determined the oral bioavailability of trovafloxacin (CP-99,219) in healthy volunteers under fasted and fed conditions. In a randomized, two-way crossover study, 12 fasting subjects received two 100 mg tablets of trovafloxacin and an equivalent dose of alatrofloxacin (CP-116,517), administered by i.v. infusion over 1 h. Alatrofloxacin, the L-Ala-L-Ala prodrug of trovafloxacin, is rapidly converted in the body to trovafloxacin. After the oral dose of trovafloxacin, the mean Cmax and AUC were 2.2 mg/L and 30.4 mg x h/L, respectively. After the infusion of alatrofloxacin, the Cmax and AUC of trovafloxacin were 3.2 mg/L and 34.7 mg x h/L, respectively. The mean T(1/2) after both treatments was about 11 h. The mean Cl and Vd(ss) of trovafloxacin after the infusion of alatrofloxacin were 1.32 mL/min/kg and 1.13 L/kg, respectively. The mean oral bioavailability of trovafloxacin was estimated to be 87.6% (range 64.8-122.1%). Another randomized, open, three-way crossover study was conducted in 12 healthy male volunteers to investigate the effect of food in the gastrointestinal tract on the bioavailability of trovafloxacin. Each subject received three 100 mg tablets after fasting overnight (treatment A) or after a standard breakfast (treatment B), or 300 mg as oral aqueous suspension after fasting overnight (treatment C). Mean Tmax after treatment B occurred 2.2 h later (3.6 h vs 1.4 h) than after treatment A. Mean Cmax and AUC were 2.3 and 2.6 mg/L and 38.2 and 39.5 mg x h/L after B and A, respectively. About 5% of the administered dose was recovered unchanged in the 24 h urine sample after all three treatments. Thus, the food reduced mean Cmax by 12% but had no appreciable effect on mean AUC. The mean bioavailability of trovafloxacin administered as treatment regimen B was 96.6% relative to that of treatment A. The respective mean bioavailabilities of trovafloxacin as treatments B and A were 91.3% and 94.5% respectively of that of treatment C. The results of these studies indicate that trovafloxacin has good oral bioavailability and that the ingestion of food is unlikely to have a clinically significant effect on the bioavailability of trovafloxacin. PMID- 9222077 TI - Effect of Maalox and omeprazole on the bioavailability of trovafloxacin. AB - To determine the effect of the concurrent administration of Maalox and omeprazole in the bioavailability of trovafloxacin (CP-99,219), an open, placebo-controlled, randomized, four-way crossover study was conducted in 12 healthy male volunteers. Each received treatments of three 100 mg trovafloxacin tablets in the morning 30 min after 30 mL of Maalox (A), 30 min after placebo (B), 2 h before 30 mL of Maalox (C) and 2 h after 40 mg of omeprazole (D). For treatments A and C, Maalox was also given at 22.00 h the night before the study day, 1 and 3 h after meals and at bedtime on the study day. For B and D, placebo and omeprazole, respectively, were also given at 22.00 h the night before the study day. After treatments A and C, mean area under the curve (AUC) was reduced by 66% and 28% (14.2 and 30.2 mg.h/L), respectively, and mean T(1/2) declined by 33% and 31% (8.3 and 8.5 h), respectively, relative to the values after B (42.1 mg.h/L; 12.4 h). The mean Kel-corrected relative bioavailabilities for A and C were 50% and 104%, respectively, suggesting a large reduction in the initial absorption of trovafloxacin with A. Treatment D had no appreciable effect on mean T(1/2) but mean AUC and Cmax were reduced by 18% and 32%, respectively, relative to B. The mean relative bioavailability after D was 82%. We conclude that the concurrent administration of trovafloxacin and aluminium- and magnesium-containing antacids should be avoided but that co-administration with omeprazole is unlikely to have a clinically significant effect on the extent of absorption of the antibiotic. PMID- 9222078 TI - Computational and numerical methods for bioelectric field problems. AB - Fundamental problems in electrophysiology can be studied by computationally modeling and simulating the associated microscopic and macroscopic bioelectric fields. To study such fields computationally, researchers have developed a variety of numerical and computational techniques. Advances in computer architectures have allowed researchers to model increasingly complex biophysical systems. Modeling such systems requires a researcher to apply a wide variety of computational and numerical methods to describe the underlying physics and physiology of the associated three-dimensional geometries. Issues naturally arise as to the accuracy and efficiency of such methods. In this paper we review computational and numerical methods of solving bioelectric field problems. The motivating applications represent a class of bioelectric field problems that arise in electrocardiography and electroencephalography. PMID- 9222079 TI - Biodegradable polymeric device for sustained intravitreal release of ganciclovir in rabbits. AB - PURPOSE: A scleral plug made of biodegradable polymer implanted at the pars plana was evaluated to determine its ability to control the intravitreal release of ganciclovir. METHODS: Scleral plugs containing 25% ganciclovir were prepared with poly(lactic-glycolic acid) (molecular weight, 121 kDa). The release of ganciclovir was evaluated in vitro by spectrophotometry. In vivo intravitreal ganciclovir concentrations were measured by high performance liquid chromatography following plug implantation in pigmented rabbits. The biocompatibility of the device was determined by indirect ophthalmoscopy, electroretinography, and light and electron microscopy. RESULTS: The in vitro study showed that the plug released ganciclovir throughout a 10-week period. The in vivo study demonstrated that the plugs maintained the drug concentration in the vitreous in a therapeutic range adequate to treat cytomegalovirus (CMV) retinitis for 12 weeks. No significant retinal toxicity was observed. CONCLUSIONS: This study demonstrated that this drug delivery system can potentially be useful to treat CMV retinitis. PMID- 9222080 TI - Real-time confocal microscopic observations on human corneal nerves and wound healing after excimer laser photorefractive keratectomy. AB - PURPOSE: Corneal wound healing after excimer laser photorefractive keratectomy (PRK) passes through a series of characteristic stages which have earlier been defined by means of histological, histochemical, and biochemical approaches. We investigated the potential of confocal microscopy to verify morphological changes in human corneas in vivo after PRK. METHODS: Ten corneas of eight patients that had earlier undergone PRK were examined at different postoperative time points (7 days-34 months). One of the PRK patients was examined sequentially three times. Three additional corneas, which had earlier undergone corneal grafting surgery and then were subjected to excimer laser photoastigmatic keratectomy (PARK), were studied as well. Seven healthy untreated corneas served as controls to define the normal morphology of human cornea. A tandem scanning confocal microscope (TSCM) was used to generate real-time images of the corneas on an S-VHS videotape. The images were either digitized and further processed or the individual video frames were produced with a video printer. RESULTS: Seven days post-PRK in vivo confocal microscopy revealed the presence of morphologically immature surface epithelial cells. Delicate nerves, activated keratocytes and deposition of extracellular light-reflecting scar tissue were perceived. The epithelium appeared normal one month post-PRK. Ongoing activation of the anterior stromal keratocytes along with extracellular scar tissue were detected. We also observed increasing numbers of regenerating subepithelial nerve leashes with somewhat twisted pattern. Highly reflective, presumably activated keratocytes were no longer detected 6-7 months post-PRK. Hypercellularity with scar tissue could still be found up to 30 months post-PRK. Only one cornea examined 34 months post-PRK showed normal keratocyte morphology and recovery of the anterior stroma. However, the morphology of subepithelial nerves was still somewhat abnormal. The two corneal grafts examined 11 or 32 months post-PARK exhibited a normal-appearing epithelium but considerable stromal hypercellularity and extracellular scar deposition. The subepithelial nerves were poorly regenerated in one eye and fairly well detectable in the other. The third graft examined 15 months post-PARK revealed the presence of enlarged surface epithelial cells and dense stromal scarring but no nerves. CONCLUSION: TSCM clinically confirms the earlier histological data on healing of excimer laser wounds. It offers a distinct improvement in the assessment of excimer laser-treated corneas, as it enables cellular details and nerves to be perceived in vivo. In addition the thickness of the stromal scar can be be measured for e.g. planning of phototherapeutic keratectomy. PMID- 9222081 TI - Abnormal naive and memory T lymphocyte subsets in the peripheral blood of patients with uveitis. AB - PURPOSE: To better define the role of lymphocytes in the pathogenesis of uveitis, we studied the expression of memory and naive cell markers on T lymphocytes from peripheral blood. METHODS: Surface antigens on T lymphocytes obtained from peripheral blood of 27 patients with uveitis, including 12 patients with Behcet's disease (BD), 7 patients with Vogt-Koyanagi-Harada disease (VKH), and 8 patients with idiopathic uveitis (IU), were detected by three-color flow cytometric analysis. Lymphocytes from 14 age-matched healthy control subjects were similarly evaluated. RESULTS: The percentage of T lymphocytes that were CD4+CD29+ lymphocytes (memory cells) was high in all patients with uveitis, while that of CD4+CD45RA+ lymphocytes (naive cells) was lower in patients with BD and VKH, although the difference was not statistically significant. The percentage of CD29+ cells within CD3+CD4+ cell population was significantly higher in patients with BD and VKH than in the controls (p < 0.01), and the percentage of CD45RA+ cells was significantly lower in BD patients than in controls (p < 0.01). The T lymphocyte subsets in patients with IU were similar to the controls. CONCLUSIONS: These results show an abnormal distribution of T lymphocytes in patients with uveitis associated with an underlying systemic disease. PMID- 9222082 TI - Retrovirus-mediated transfer of the suicide gene into retinal pigment epithelial cells in vitro. AB - PURPOSE: Human retinal pigment epithelial (HRPE) cells are a major cell component in proliferative vitreoretinopathy (PVR) membranes. We investigated the feasibility of killing HRPE cells by retroviral-mediated transfer of the herpes simplex virus-thymidine kinase (HSV-tk) gene, also known as the suicide gene, into HRPE cells followed by ganciclovir treatment, and to study the so-called bystander effect. Such a treatment plan might serve as a possible therapy for PVR. METHODS: Transduction efficiency was determined using retroviral vectors encoding the beta-galactosidase reporter gene. To evaluate the efficacy of suicide gene therapy. HRPE cells were transduced with retroviral vectors encoding the HSV-tk gene (G1TkSvNa), with empty vectors or without vectors, and were treated with 5 micrograms/ml ganciclovir. Sensitivity of HSV-tk positive HRPE cells to various concentrations of ganciclovir was evaluated. To demonstrate the bystander effect, HSV-tk positive cells were cultured with HSV-tk negative cells at varying proportions. RESULTS: Transduction efficiency in vitro was 15.1 +/- 4.8%. Cell growth was significantly inhibited after transduction with G1TkSvNa followed by ganciclovir treatment (P < 0.01). Ganciclovir showed dose- and time dependent cytotoxicity only on HSV-tk positive cells. The concentration that resulted in 50% inhibited was 0.1 micrograms/ml. In terms of the bystander effect, after ganciclovir treatment the viability of co-cultured cells decreased with increasing populations of HSV-tk positive cells. CONCLUSIONS: HRPE cells were successfully transduced with the HSV-tk gene via retroviral vectors and displayed a strong bystander effect after treatment with ganciclovir. These results suggest that retrovirus-mediated suicide gene therapy might be a feasible treatment strategy for PVR. PMID- 9222083 TI - Drug distribution in the vitreous humor of the human eye: the effects of intravitreal injection position and volume. AB - PURPOSE: The purpose of this study was to determine how the position and volume of an intravitreal injection affect the distribution and elimination of drug from the vitreous humor. METHODS: A mathematical model that had been developed and used previously to study drug distribution in the vitreous humor of the rabbit eye was modified to match the physiology of the human eye. Fluorescein and fluorescein glucuronide were used as the model compounds for these studies. Four extreme injection locations were considered: a central injection, an injection displaced towards the retina, an injection displaced towards the lens, and an injection displaced toward the hyaloid membrane. Injections containing an equal mass of drug dissolved in volumes of either 15 microL or 100 microL were compared. RESULTS: The location of an intravitreal injection was found to have a substantial effect on elimination and distribution in the vitreous. Peak concentrations at different vitreous locations varied by over three orders of magnitude, depending on the injection location. The mean concentration of drug remaining in the vitreous 24 hours after the intravitreal injection varied by up to a factor of 3.8, depending on the injection location. Changing the volume of the injection from 15 microL to 100 microL dampened the effects of the initial injection location; however, meant concentrations at 24 hours still varied by up to a factor of 2.5. CONCLUSIONS: Careful control of the conditions of an intravitreal injection could reduce treatment variability, improve bioavailability, and reduce the possibility of retinal toxicity. PMID- 9222084 TI - Cytokine expression in the alkali-burned cornea. AB - PURPOSE: This study investigated the cytokine expression profile in alkali-burned mouse corneas, in order to elucidate the mechanisms of corneal damage and repair. METHODS: The cytokines expressed in alkali-burned corneas were identified by polymerase chain reaction (RT-PCR), then quantitated using ELISA. Based on the ELISA results, immunohistochemical analyses were performed to localize cytokine expression. RESULTS: Among the ten cytokines examined, IL-1 (IL-1 beta), IL-6, IL 10 and TNF-alpha mRNA were expressed in alkali-burned corneas. Quantitation revealed that IL-alpha and IL-6 were strongly induced in the early stages of alkali burn, peak production of IL-1 alpha (53.2 pg/cornea) and IL-6 (23.6 pg/cornea) occurring at days 3 and 7, respectively. The production of IL-10 and TNF-alpha was not significantly elevated during the 42 day period after burn. Immunohistochemical analyses revealed that both IL-1 alpha and IL-6 were mainly localized in regenerating epithelial basal cells. CONCLUSIONS: IL-1 alpha and IL 6 levels in the cornea are dramatically elevated in the regenerated epithelium during the early stages of alkali burn, and may play an important role in associated corneal damage and repair. PMID- 9222085 TI - Keratan sulphate in the trabecular meshwork and cornea. AB - PURPOSE: A study was made of the distribution of keratan sulphate in the human anterior chamber. METHODS: The monoclonal antibody, 5-D-4, was used in immuno electron microscopy to visualise keratan sulphate distribution in the anterior chamber of 16 normal eyes, 7 Fuchs' dystrophy corneas, and a macular dystrophy cornea. RESULTS: Keratan sulphate was detected in normal human aqueous humour and also on the surface of trabecular cells in the uveal meshwork. Normal corneal stroma showed an increase in keratan sulphate labelling from anterior to posterior, with marked labelling in the posterior region of Descemet's membrane. The apical surface of the corneal endothelium labelled positively, but showed considerable variation in the level of labelling from cell to cell. The macular dystrophy cornea had the classic histopathological features of a type I case, including a highly abnormal Descemet's membrane. No keratan sulphate was detected in the macular dystrophy patient's corneal stroma or serum. The Fuchs' endothelial dystrophy corneas showed a normal distribution of keratan sulphate labelling in the stroma. The Fuchs' endothelial cells labelled for keratan sulphate but were highly abnormal in appearance, often exhibiting long filopodia and appearing to be actively migrating. CONCLUSIONS: This work has shown that keratan sulphate has a much wider distribution than was previously believed. The detection of keratan sulphate on the trabecular and endothelial cell surfaces also suggests a possible role for this molecule in cell adhesion and/or migration. PMID- 9222086 TI - Effects of endothelin-1 (ET-1) on ocular hemodynamics. AB - PURPOSE: There is evidence from in vitro and animal data that endothelin-1 (ET-1) plays an important role in ocular blood flow. The aim of the present study was to investigate the effect of systemic ET-1 administration on ocular circulation in healthy subjects. METHODS: In a double blind, placebo-controlled, randomized, 2 way cross over study in 10 healthy male subjects, we administered stepwise, increasing doses of ET-1 (0 (saline), 1.25, 2.5 and 5.0 ng/kg/minutes; 20 minutes per dose (level) or placebo. Blood flow velocity in the ophthalmic artery as well as ocular fundus pulsations in the macula and the optic disc, and systemic hemodynamic parameters were measured. RESULTS: ET-1 dose-dependently reduced fundus pulsations in the macula (maximum effect -12 +/- 2% versus baseline; p < 0.001 versus baseline and placebo) and the optic disc (maximum effect: -19 +/- 5% versus baseline; p < 0.001 versus baseline and placebo), but did not affect blood flow velocity in the ophthalmic artery or systemic hemodynamics. CONCLUSIONS: Endothelin-1 reduces pulsatile blood flow in the choroid and the optic disc at doses which do not affect systemic hemodynamics or flow velocity in the ophthalmic artery. These results indicate that ocular circulation is particularly sensitive to changes in local ET-1 concentration and confirms the hypothesis that ET-1 may play a role in ocular vascular diseases. PMID- 9222088 TI - Lysophosphatidic acid stimulates proliferation of human retinal pigment epithelial cells. AB - PURPOSE: Proliferative vitreoretinopathy (PVR) can arise from an exaggerated wound-healing response by retinal pigment epithelial (RPE) cells. Lysophosphatidic acid (LPA) is a simple phospholipid, which is secreted by cells, activates G protein-coupled receptors, and appears to contribute to wound healing in other tissues. The present study examined the effects of LPA on three aspects of the behavior of cultured human RPE cells that are important in the pathogenesis of PVR: proliferation, chemotaxis, and contraction. METHODS: Human RPE cells were harvested from donor eyes and cultured using standard culture techniques. Proliferation was assessed by counting cells, cell migration with a modified Boyden chamber, and contraction by seeding RPE cells in a collagen cell. RESULTS: LPA (10 microM) induced RPE cell proliferation and weak chemotaxis, but no gel contraction. RPE cell proliferation increased in a dose-dependent manner from 0.1-100 microM LPA. Consistent with LPA actions at a receptor, an LPA analogue, lysophosphatidylcholine (LPC), was much less effective than LPA in stimulating proliferation and the proliferative response was blocked by pertussis or cholera toxin. Phosphatidic acid (PA) induced a similar proliferative response as LPA. CONCLUSION: These suggest that LPA can potently stimulate RPE cell proliferation via activation of a G-protein coupled receptor. LPA, which can be released by thrombin-activated platelets and growth factor-activated fibroblasts, might, therefore, play a role in the development of PVR. PMID- 9222087 TI - Cidofovir transport in the pigmented rabbit conjunctiva. AB - PURPOSE: To characterize cidofovir (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl) cytosine) transport in the pigmented rabbit conjunctiva and to evaluate the formulation influence on its transport. METHODS: The excised pigmented rabbit conjunctiva was mounted in the modified Ussing chamber. Cidofovir transport was initiated by applying 3H-cidofovir to the donor compartment and assayed by measuring the radioactivity accumulated in the receiver fluid over 180 min. RESULTS: Cidofovir flux in the mucosal-to-serosal direction increased proportionally with drug concentration over the 0.01 to 1 mM range. Cidofovir transport (0.01 mM) at 37 degrees C in the mucosal-to-serosal direction was not significantly different from that in the opposite direction or from that at 4 degrees C. Hypotonicity (80 mOsm/kg), 0.5% EDTA, and 0.0125% benzalkonium chloride increased the apparent permeability coefficient of cidofovir 3, 21, and 49 times, respectively. This was accompanied by a corresponding 43%, 86%, and 96% reduction in the transconjunctival electrical resistance over 180 min. The reduction in transepithelial electrical resistance elicited by hypotonicity was reversible. There was a good correlation between apparent permeability coefficient and the transconjunctival conductance, suggesting that cidofovir may undergo paracellular passive diffusion in the conjunctiva. CONCLUSION: Cidofovir transport in the rabbit conjunctiva may be via paracellular passive diffusion. Formulation changes may improve cidofovir absorption from the conjunctival route. PMID- 9222089 TI - Antiproliferative effect of retinoic acid in 1% sodium hyaluronate in an animal model of PVR. AB - PURPOSE: To determine the antiproliferative activity in intravitreous retinoic acid (RA) dispersed in 1% sodium hyaluronate (HA). METHODS: Six groups of pigmented rabbits underwent gas-compression vitrectomy. Four days later, gas/HA or gas/balanced salt solution (BSS) exchange (1.0 m1) was performed in all rabbits. Groups A (n = 10) and B (n = 5) received intravitreous RA dissolved in 0.01 m1 of ethanol and dispersed in 1% HA (10 and 15 micrograms RA/m1, respectively). Group C (n = 10) received intravitreous RA dissolved in ethanol and dispersed in BSS (10 micrograms RA/m1). Groups D (n = 5) and F (n = 4) received 1 m1 of HA with ethanol; group E (n = 5) received 1 m1 of HA without ethanol. All groups except group F also received homologous fibroblasts and autologous, platelet-rich plasma intravitreously. The eyes were examined ophthalmoscopically for 1 month. Proliferative vitreoretinopathy (PVR) findings were graded according to the classification of Fastenberg et al. all group F eyes also were examined by light and electron microscopy. RESULTS: RA in HA lessened PVR progression within 1 month when compared with HA injection controls and within 2 weeks when compared with the RA in BSS treatment group (both, p < 0.05). NO specific change attributable to ethanol was observed histopathologically. CONCLUSION: RA dissolved in ethanol and dispersed in HA could be useful to treat PVR when silicone oil is unnecessary. PMID- 9222090 TI - A low conductance chloride channel in the basolateral membranes of the non pigmented ciliary epithelium of the rabbit eye. AB - PURPOSE: Chloride efflux plays an important role in aqueous humor production. Chloride currents have been described in bovine non-pigmented ciliary epithelium (NPE), in transformed cultured human NPE, and in bovine volume-activated chloride channels described in the latter. It is the basolateral membranes of the NPE of ocular ciliary processes that comprise the exit pathway for the process of aqueous secretion performed by the double syncytial layer of ciliary epithelium, however. Therefore, we studied both cell-attached, and, excised, inside-out patches from basolateral membranes of the NPE. METHODS: Cell-attached and cell free excised patches were formed from the basolateral membranes of NPE and single channel currents recorded with a Dagan 3900A patch clamp amplifier. RESULTS: A low conductance channel of 14 pS was observed and recorded in 30% of cell attached patches and in 35% of excised inside out patches under symmetrical conditions (160 mM chloride). This channel displayed a nearly linear current voltage relationship, with an open probability that was not voltage-dependent. The channel was chloride-selective: N-methyl-D-glucamine (NMDG) used as cation did not alter the current profile nor the reversal protein. Further, with inside out patches, the reversal potential was close to zero (0.3 +/- 0.5 mV (10) in symmetrical (160 mV) chloride, but shifted to -32.3 +/- 0.5 mV (5) when the concentration of chloride in the bathing solution was reduced to 40 mM while the recording pipette was held at 160 mM. This value approaches the theoretical equilibrium potential of chloride for these conditions. The channel anion permeability sequence was: I- > NO3- > or = Br- > Cl- > gluconate- approximately equal to aspartate-. Three different chloride-channel blockers inhibited the channel activity. CONCLUSION: A low conductance channel, selective for chloride, and, modulated by beta adrenergic and VIP stimulation, based on it sensitivity to exogenously added cAMP, ATP and the catalytic subunit of PKA, is present in the exit basolateral membranes of rabbit NPE and contributes to the process of aqueous humor formation. PMID- 9222091 TI - Beta A3/A1 crystallin from human cataractous lens contains an intramolecular disulfide bond. AB - PURPOSE: To develop an experimental approach that can identify amino acid sequences containing cysteine residues involved in disulfide bonding during cataractogenesis of the human lens. METHODS: Total proteins from cataractous and normal human lenses were solubilized anaerobically, followed by carboxy methylation of free sulfhydryl groups with iodoacetate. Carboxymethylated proteins were partially purified by reverse phase chromatography, then subjected to lys-C endoprotease digestion. Using reverse phase chromatography, each digest was resolved in the presence and absence of dithiothreitol (DTT), to identify peptides that were linked by disulfide bonds. These peptides were further characterized using a combination of Edman degradation and mass spectrometry. RESULTS: The reverse phase chromatography profiles of lys-C peptides from proteins of normal lenses were very similar in the presence and absence of dithiothreitol, while identical analysis of proteins from cataractous lenses demonstrated the presence of a lys-C peptide that corresponded to residues 163 193 of human beta A3/A1, with cysteine 170 and cysteine 185 linked via an intramolecular disulfide bond. CONCLUSIONS: Reverse phase chromatography of complex mixtures of lens protein digests, in the presence and absence of dithiothreitol, provides a rapid method of identifying sequences involved in disulfide bonding. The results of this analysis using the endoprotease lys-C have demonstrated that beta A3/A1 crystallin from cataractous lenses contains an intermolecular disulfide bond involving cysteine residues 170 and 185. PMID- 9222092 TI - The effect of carbon dioxide on oxygen-induced retinopathy in the neonatal rat. AB - PURPOSE: Hypercarbia has been suggested as a risk factor for retinopathy of prematurity. We investigated the effect of raised inspired carbon dioxide on oxygen-induced retinopathy in the neonatal rat. METHODS: Newborn rats raised in expanded litters (n = 25 each) were exposed to cycles of hyperoxia (80% O2) and hypoxia (10% O2 for 7 days, followed by room air recovery for 5 days. During cyclic oxygen exposure, 3 litters (n = 75) were exposed to 10% CO2 (PaCO2 78 mm Hg +/- 6; mean +/- SD) and 3 litters (n = 75) were exposed to 0.2% CO2 (PaCO2 45 mm Hg +/- 7). Animals were sacrificed on day 13 and retinae were analyzed using fluorescein perfusion and ADPase staining techniques. RESULTS: Neovascularization occurred in 85% of rats exposed to high CO2 compared to 52% of rats exposed to low CO2 (p = 0.001). The severity of neovascularization, in clock hours, was also greater in the rats exposed to high CO2 (p < 0.001). CONCLUSIONS: Exposure to high CO2 results in an increased incidence and severity of neovascularization in a rat model for oxygen-induced retinopathy. Our results support the suggestion that hypercarbia may be a risk factor for retinopathy of prematurity. PMID- 9222093 TI - Lineage study of degenerating photoreceptor cells in the rd mouse retina. AB - PURPOSE: A retroviral marker was used to label daughter cells arising from individual neuroblasts in the rd mouse retina, in order to investigate the hypothesis that a clonal relationship exists among degenerating photoreceptor cells. METHODS: On the day of birth, a single injection of retrovirus with a lac Z (beta-galactosidase) reporter construct was injected into the retina in the vicinity of the subretinal space. Descendants of single neuroblasts were identified histochemically by examining the retinas at P14 (postnatal day 14). Light and electron microscopic studies were used to identify the retrovirally induced marker beta-galactosidase using Bluo-gal dye. Double-labeling of degenerating cone cells was accomplished by taking 100 microns vibratome sections of retrovirally-injected eyes and using either FITC-PNA or HRP-PNA to visualize clusters of degenerating cones as well as Bluo-gal labeled clones of photoreceptor cells on the same tissue section. RESULTS: A relatively large number of clones of primarily photoreceptor cells were observed in the peripheral retinas of both normal and rd mice. In a few cases in the rd, photoreceptor cells in a given clone consisted of both PNA- and Bluo-gal-labeled cells as well as of only Bluo-gal-labeled cells. CONCLUSION: These results suggest that during the period of intense cell death in the rd retina, a single dying photoreceptor cell can be surrounded by photoreceptors (either rods or cones) from the same clone that appear morphologically normal without evidence of degeneration. PMID- 9222095 TI - Photoreceptor rim protein: partial sequences of cDNA show a high degree of similarity to ABC transporters. AB - PURPOSE: To isolate and sequence cDNA for bovine rim protein, a large membrane bound glycoprotein found in photoreceptor outer segments. METHODS: Bovine rim protein was N-terminally sequenced (22 residues) and fragments were prepared by partial proteolysis. Two internal sequences of 21 and 18 amino acid residues were obtained from 35 kDa and 32 kDa fragments, respectively. Sense and anti-sense oligonucleotide primers were constructed, based on the peptide sequences derived from the 35 kDa and 32 kDa fragments, respectively, and the polymerase chain reaction (PCR) was used to amplify a 150 bp sequence from bovine retinal cDNA. RESULTS: The amplified sequence coded for the remainder of the peptide sequence determined from the 35 kDa fragment, which was not present in the primer, confirming that it was derived from the rim protein. The 150 bp sequence was translated to give a 50 amino acid peptide. Part of this peptide was compared with Dna sequence databases using the TFastA program, which found 94.6% identity with an EST derived from human retina and 86.1% identity to the mouse abc1 transporter. CONCLUSIONS: It is proposed that rim protein is a member of the ATP transporter family of proteins. It may be involved in transport of molecules involved in visual transduction across the photoreceptor disk membrane. PMID- 9222094 TI - Changes in the optic disc over a five-year interval: the Beaver Dam Eye Study. AB - PURPOSE: To determine the relationship of change in vertical optic disc cupping to change in intraocular pressure over a five-year interval. METHODS: Non simultaneous stereoscopic photographs were taken of optic discs of participants in the baseline and follow-up examinations of The Beaver Dam Eye Study cohort. Optic discs and cups were measured and other disc features were graded according to a standard protocol by trained graders. Intraocular pressures were measured by Goldmann applanation tonometry. RESULTS: Change in pressure was significantly associated with change in vertical cup-to-disc ratio. Incident disc hemorrhage, flattened temporal rim, notching, cup reaching disc margin, and undercutting were not significantly associated with change in intraocular pressure. CONCLUSION: Change in intraocular pressure in this adult population was associated with increased optic disc cupping. This finding, if confirmed, would lend support to the practice of periodic follow-up of older adults who have shown changes in their intraocular pressure. PMID- 9222096 TI - Analysis of variation in results of CD34+ hematopoietic progenitor cell enumeration in a multicenter study. AB - A workshop was held in The Netherlands and Belgium with the aim of investigating whether or not the use of a standard protocol vs. local protocols for flow cytometric enumeration of CD34+ hematopoietic progenitor cells would reduce interlaboratory variation. The standard protocol consisted of a three-color, whole-blood staining technique based on fluorescein isothiocyanate (FITC)-labeled CD34, and phycoerythrin (PE)-labeled CD14 and CD66e monoclonal antibodies (reactive with monocytic and myeloid cells, respectively), followed by erythrocyte lysis, washing, fixation, and selection of nucleated cells during data acquisition on the basis of their positivity for LDS-751 (staining DNA and RNA). Data analysis guidelines included the elimination of nonspecific antibody binding by monocytes and myeloid cells by gating on the CD14-, 66e- cells, followed by setting a window on a CD34 vs. sideward light scatter (SSC) plot around the CD34+, SSClow cells. The FITC-labeled isotype control was analyzed with the same gate and window settings, and the false-positive events were subtracted from CD34 result. Four samples (i.e., peripheral blood and apheresis product from two patients) were sent out. Results were received on patient 1 (2) from 36 (38) laboratories. Data obtained by 24 (26) laboratories after correct application of the standard protocol revealed that the median percentage of CD34+ cells of the four samples ranged between 1.1% and 3.7% and the CVs between 18% and 30%. Incorrect performance of the standard protocol by 12 laboratories, mainly resulting from gating errors, yielded a larger variation (CVs ranging between 50% and 82%). CD34 enumeration using local protocols by 29 (34) laboratories yielded median percentage of CD34+ cells ranging between 1.2% and 3.9% and CVs ranging between 34% and 106%. We conclude that correct application of this standard protocol was effective in reducing the interlaboratory variation in percentage of CD34+ cells assessments. PMID- 9222097 TI - Analysis of IL-2 and IL-6 binding to peripheral blood lymphocytes in Graves disease: relationship with disease activity. AB - Growing evidence points to the involvement of cytokines in the pathogenesis of some autoimmune diseases. To investigate the possible role of interleukin 2 (IL 2) and interleukin 6 (IL-6) on the pathogenesis of Graves disease (GD), the binding of both exogenous IL-2 and IL-6 and the expression of the IL-2 receptor subunit p55 (CD25) were evaluated by flow cytometry in peripheral blood T and B cells from 70 GD patients, in different states of the disease, and from 19 age- and sex-matched healthy volunteers. Serum levels of total T3 and T4, of free T4, and of anti-TSH receptor antibodies were also simultaneously determined. All GD patients displayed significantly increased numbers of B cells bound to IL-2. Hyperthyroid untreated GD patients had significantly higher numbers of T and B cells expressing the IL-2 receptor subunit p55 as compared to euthyroid patients in long-term remission. In addition, serum anti-TSH receptor antibody levels were directly correlated with the absolute numbers of T cells bound to IL-2 (r = 0.565, P < 0.05) and to IL-6 (r = 0.653), P = 0.02) in the hyperthyroid untreated patients, but not in long-term remission euthyroid GD patients or in patients treated with methimazole. The serum levels of total T3 and free T4 were significantly correlated with the absolute numbers of circulating T cells binding IL-2 (r = 0.720, P < 0.01 and r = 0.783, P < 0.002, respectively) as well as with the absolute numbers of circulating T cells binding IL-6 (r = 0.671, P < 0.02 and r = 0.626, P < 0.02, respectively). The serum levels of total T3 were also correlated with both the absolute numbers of B cells binding to IL-2 (r = 0.586, P < 0.05) and to IL-6 (r = 0.757, P < 0.001). These findings suggest that IL-2 and IL-6 may play a role in the pathogenesis of GD. PMID- 9222098 TI - Comparative analysis of whole blood lysis methods for flow cytometry. AB - We performed a parallel evaluation of six whole blood lysis methods comparing light scatter and quantitative fluorescence intensity based on quantitative flow cytometry, of selected lymphocyte subsets and CD34+ cells. Leukocytes prepared with FACS Lysing Solution (BDIS), Immunolyse (Coulter) and Optilyse B (Immunotech) consistently gave lower forward scatter values than those prepared with ACK (BioWhitaker), Ortho-mune (Ortho) and ImmunoPrep (Coulter). Debris, defined as CD45 negative events with the threshold off, accounted approximately 80% of all events with ACK and Ortho-mune. The other lysing methods consistently yielded less debris (approximately 50%) with Immunolyse generating only approximately 16% debris. Optilyse and FACS lyse consistently displayed the lowest percentage of lymphoid cells (CD45+/CD14-) in the three part differential. The percentage of CD3+, CD20+, CD5+, and CD16/CD56+ cells was consistent with all methods but CD4 and CD8 determinants showed inconsistent variation with ACK and Ortho-mune. In addition, the fluorescence intensity of CD14 PE and CD8 PE staining was markedly decreased on cells prepared with ImmunoPrep. Finally, the clearest separation of CD34+ cells was observed with ACK and Ortho-mune. Our data demonstrate that the method used for red cell lysis can have definite impact on immunophenotyping and selected methods appear to be more suitable for specific applications. PMID- 9222099 TI - Comparative analysis of different permeabilization methods for the flow cytometry measurement of cytoplasmic myeloperoxidase and lysozyme in normal and leukemic cells. AB - Using a direct one-color (fluorescein isothiocyanate; FITC) staining method with a Facscan flow cytometer, we evaluated the intracellular expression of two granular constituents of myeloid cells [myeloperoxidase (MPO) and lysozyme] on leukemic cells from 21 patients with acute myeloid leukemia (AML), and 6 patients with acute lymphoblastic leukemia (ALL). Three different permeabilization techniques were used [FACS Lysing Solution (FLy), B.Dis; Ortho-PermeaFix (OPF); Fix and Perm (F&P), Caltag] prior to monoclonal antibody (McAb) staining, in order to verify the specificity and the sensitivity of the three labelling methods towards the two model antigens. Peripheral blood cells from 15 healthy subjects and Ortho Absolute Control served as controls. Data were expressed as percentage of positivity, net fluorescence intensity, ratio between mean fluorescence intensity (MFI) of positive cells and that of isotypic controls (P/N ratio; evaluated in both geometric and arithmetic scale), and, in 12 representatives cases (7 AML, 5 normal samples), in the form of both molecules of equivalent soluble fluorochromes (MESF) and antibody binding capacities (ABC). As far as the antigenic expression of MPO and lysozyme in normal samples is concerned, F&P resulted, in our hands, in the most specific and sensitive staining, followed by FLy solution and OPF, which showed positivity for MPO, and, to lesser extent, for lysozyme in a considerable manner of lymphocytes (means 64% and 54%, respectively, for OPF and FLy; range of ABC/cell: 0.9-5.2 x 10(3)) obtained from healthy subjects. With the reference F&P permeabilizing solution, 90% and 80% of FAB M1-M5 cases were found to be positive for MPO and lysozyme, respectively. However, M1, M2, and M3 AML FAB (French-American-British) subvarieties were characterized by a brighter expression for MPO (mean ABC/cell: 89 x 10(3)) than that of lysozyme (mean ABC/cell: 12.5 x 10(3D)), whereas blast cells from patients with M5a FAB subtypes showed higher levels of lysozyme (mean ABC/cell: 65 x 10(3)) than that of MPO (mean ABC/cell: 0.1 x 10(3)). One of five cases of FAB MO AML showed a dull positivity for MPO-7 McAb. Patients with ALL were MPO and lysozyme negative using both F&P and FLy reagents, although a certain degree of positivity was documented in some cases with OPF. Taking these data together, it can be stated that the use of anti-MPO McAbs may be of great value for the diagnosis and monitoring of acute leukemia and, along with lysozyme McAb, can provide useful information in the distinction of myeloid from monocytic leukemias and in the lineage assignment of apparently biphenotypic forms. However, the methodology used for the detection of these myeloid-associated antigens is critical for a correct interpretation of cytofluorimetric data and should be taken into account when evaluating data coming from multicenter trials dealing with leukemias. A standardization of cytofluorimetric analysis of intracellular antigens is needed in order to improve the reproducibility and comparability of results in multicenter studies. PMID- 9222100 TI - An artificial intelligent diagnostic system on differential recognition of hematopoietic cells from microscopic images. AB - Despite their advantages, none of the automated white blood cell differentiated counters have replaced the conventional microscopic evaluations of blood and bone marrow slides by hematologists. We have analyzed the smears of 39 patients and 8 control subjects to develop an artificial expert system that recognizes 16 different types of nucleated hematopoietic cells during the stages of differentiation. A charge coupled television camera and a special frame grabber were used for data acquisition, and 247 nucleated cell images were transferred from a microscope to an IBM 386 computer to be processed. One hundred sixty-five and 82 of these images were used for training and testing, respectively. Our system is composed of image processing and analysis (enhancement, thresholding/smoothing, edge detection), pattern recognition (feature extraction and classification with supervised artificial neural network), and expert system development. Image processing and analysis were used to obtain 13 cellular features to be used as the input parameters (neurons) of the artificial neural network. A supervised artificial neural network (back-propagation learning algorithm) was used in the classification of 16 different cells (output neurons of the neural network), which is the second step of pattern recognition. A confusion matrix has been developed to compare the similarities and dissimilarities between the differential recognitions of the hematologist and the expert system. The discriminatory power of the procedure is statistically significant: Q = (N - n.K)2/N.(K - 1) = 28.2. The sensitivity and the specificity of the expert system were 71.4% and 90.9%, respectively. PMID- 9222101 TI - Reducing cellular autofluorescence in flow cytometry: an in situ method. AB - Cellular autofluorescence affects the sensitivity of flow cytometric assays by interfering with detection of low level specific fluorescence. These detection limits increase with use of protocols, such as thermocycling and fluorescent in situ hybridization (FISH), that can increase intrinsic cellular fluorescence to 5,000-20,000 fluorescein isothiocyanate (FITC) equivalents. In order to improve signal to noise ratios when using FITC labeled probes in these procedures, we employed a method using the polyanionic azo dye, trypan blue, to reduce intracellular autofluorescence. Dyes such as these are commonly used in immunofluorescent microscopy to reduce background fluorescence. By using this method, we realized an approximately 5-fold increase in signal to noise ratio (S/N) in the direct detection of RNA target probes using flow cytometry. Trypan blue aided in the resolution of dim surface antibodies, internal markers and probes, and functions to reduce background autofluorescence after thermocycling and hybridization. This technique is rapid and easily applicable for reducing intracellular autofluorescence, and can be used in single and dual color applications. PMID- 9222102 TI - A t-statistic for objective interpretation of comparative genomic hybridization (CGH) profiles. AB - An objective method for interpreting comparative genomic hybridization (CGH) is described and compared with current methods of interpretation. The method is based on a two-sample t-statistic in which composite test:reference and reference:reference CGH profiles are compared at each point along the genome to detect regions of significant differences. Composite profiles are created by combining CGH profiles measured from several metaphase chromosomes for each type of chromosome in the normal human karyotype. Composites for both test:reference and reference:reference CGH analyses are used to generate mean CGH profiles and information about the variance therein. The utility of the method is demonstrated through analysis of aneusomies and partial gain and loss of DNA sequence in a myeloid leukemia specimen. Banding analyses of this specimen indicated inv (3)(q21q26), del (5)(q2?q35), -7, +8 and add (17)(p11.2). The t-statistic analyses of CGH data indicated rev ish enh (8) and rev ish dim (5q31.1q33.1,7q11.23qter). The undetected gain on 17p was small and confined to a single band (17p11.2). Thus, the t-statistic is an objective and effective method for defining significant differences between test and reference CGH profiles. PMID- 9222104 TI - Quality control of CGH: impact of metaphase chromosomes and the dynamic range of hybridization. AB - With the recent rapid expansion in the use of the comparative genomic hybridization (CGH) technique, increased attention to quality control is essential. In the present study, we show that despite optimization and standardization of metaphase preparation techniques and the commercial availability of metaphase spreads, batch-to-batch variability of the preparations remains a significant problem. To facilitate reliable CGH analysis despite this variability, we have developed a rapid denaturation test to assess the quality of the preparations without hybridization and quantitative image analysis criteria for assuring the day-to-day quality of CGH experiments, including sensitivity, specificity, and dynamic range. Monitoring the dynamic range of the hybridizations was found to be particularly critical for achieving sensitive and reliable CGH results. This reliability can be achieved, for example, by hybridization of a green-labeled normal male DNA against red-labeled female DNA and monitoring of the green:red ratio of the X chromosome in relation to that of the autosomes. PMID- 9222103 TI - Objective aneuploidy detection for fetal and neonatal screening using comparative genomic hybridization (CGH). AB - Comparative genomic hybridization (CGH) allows entire genomes to be scanned for whole and segmental aneuploidy and thus may be an appropriate tool for the detection of clinically important abnormalities during fetal and neonatal screening. Criteria to distinguish between significant aberrations and experimental artifacts are essential for these applications. This report describes the use of a t-statistic to detect changes in CGH profiles that differ significantly from variations that occur in CGH profiles of normal samples. Eleven cell lines derived from fetal or neonatal patients were analyzed in this study. Aneuploidies in these lines included trisomies for chromosomes 13, 16, 18, and 21 and monosomy for distal 5p and tetrasomy 18p. Aneuploidy was detected in all samples by using the t-statistic, although the extent of the aneuploid region was not correctly estimated in some cases. A detailed description of the t statistic fused for making these CGH comparisons is described in a companion paper (Moore et al., Cytometry 28:183-190, 1997. PMID- 9222105 TI - Molecular mechanism controlling the incorporation of fluorescent nucleotides into DNA by PCR. AB - The efficiency and yield of incorporation of fluorescent nucleotides into DNA by polymerase chain reaction (PCR) have been investigated with linear amplification (PCR with single-stranded template and single primer). In the present study, we prepared single-stranded templates with defined sequences and used dUTP attached to the fluorescent label with linkers of different lengths. Incorporation and yield of the modified dUTP were reduced when the sequence demanded that multiple dyes be inserted at adjacent sites. The interactions between the polymerase and cyanine-labeled sites on the extending strand probably terminated the chain extension. Thus, because labeling density was increased, the yield of PCR was reduced. We also found that the interactions between the primer and dye-labeled sites on template disturb primer annealing and lead to a decrease in PCR yield. PMID- 9222106 TI - DNA binding fluorochromes as probes for histone H1-chromatin interactions in situ. AB - We have investigated using the DNA binding fluorochromes 7-aminoactinomycin (7 AAMD) and 4',6-diamidino-2-phenylindole (DAPI) as cytochemical probes for linker histone (H1)-chromatin interactions in situ. Human lymphocytes, permeabilized with digitonin, were exposed to increasing concentrations of sodium chloride to remove ionically bound H1 from the nuclei. The cells were stained to equilibrium with 1 microM 7-AAMD or 50 nM DAPI. Lymphocytes stained with 7-AAMD showed a gradual increase from 11% to 36% of HC1 treated cell fluorescence intensity when the salt concentration was increased from 0.15 to 0.7 M. The corresponding increase for DAPI was 53-68%. The 7-AAMD obviously showed higher sensitivity for H1-chromatin interactions that DAPI but had disadvantages such as high background fluorescence and an affinity that was dependent on the preparation procedure. DAPI had negligible background fluorescence, and its fluorescence intensity resembles the number of available high-affinity dye-binding sites when used at 50 nM. We conclude that both fluorochromes can be used as probes for H1-chromatin interactions in situ and that our method has a potential to provide new information on such interactions. PMID- 9222108 TI - Flow cytometric analysis of micronucleus induction in rat bone marrow polychromatic erythrocytes by 1,2;3,4-diepoxybutane, 3,4-epoxy-1-butene, and 1,2 epoxybutane-3,4-diol. AB - Automation of the analysis of micronucleus induction with flow cytometry was developed by using mouse bone marrow or peripheral blood. In the present study, we report the use of flow cytometry for the identification and quantification of micronuclei (MN) induced in rat bone marrow polychromatic erythrocytes. Three metabolites of the industrial chemical 1,3-butadiene, namely 1,2;3,4 diepoxybutane (DEB), 3,4-epoxy-1-butene (EB) and 1,2-epoxybutane-3,4-diol (diol EB), were studied in addition to mitomycin C and cyclophosphamide, which served as positive controls. DEB showed a dose-dependent increase in the frequency of MN, whereas EB was completely negative and diol-EB only weakly positive at one dose level. The effect of the positive control compounds was observed 48 h after a single injection in a dose-dependent manner. Flow cytometry was an effective method to quantitate bone marrow MN induction in the rat when density gradient separation of polychromatic erythrocytes is used. The results are compatible with the theory that oxidation of EB to the mutagenic metabolite DEB occurs at a low rate in rat bone marrow and that EB is detoxified by epoxide hydrolase and by conjugation with glutathione by glutathione transferase yielding nonmutagenic metabolites. Thus, the reported lack of MN induction by 1,3-butadiene inhalation in rat bone marrow is explained. PMID- 9222107 TI - Multivariate reconstruction of lymphocyte profiles in a two-dimensional graphical model as a tool for the investigation of lymphocyte subset distribution in health and disease. AB - Advanced multivariate data-analytical techniques are proposed to concisely represent and evaluate complex lymphocyte profiles (i.e., compound lymphocyte subset distributions) of individual subjects in easily interpretable, two dimensional, graphical correlation biplots. The lymphocyte profile of each subject is represented by its location in the model, and the score of a subject for a particular lymphocyte subset is inferred from the perpendicular projection on a rotated axis that coincides with this lymphocyte subset. Simultaneously, the model yields information about the correlation between the lymphocyte subsets. Furthermore, individuals with aberrant lymphocyte profiles can be easily identified. In case studies of 80 healthy donors and of 40 patients with multiple myeloma, 10 patients with monoclonal gammopathy of undetermined significance, and 50 age- and sex-matched healthy donors, reconstruction of the two-dimensional lymphocyte profiles from 27 flow-cytometric characterized lymphocyte subsets succeeded in representing 43% and 51% of the total information (variability) contained within the 80 x 27 (= 2,160) and 100 x 27 (= 2,700) flow cytometry measurements, respectively. It is concluded from the present studies that the correlation biplot represents a unique and powerful tool to concisely describe, represent, and analyze complex lymphocyte profiles of individual subjects and the heterogeneity in lymphocyte profiles among these subjects. PMID- 9222109 TI - Bivariate flow karyotyping in broad bean (Vicia faba). AB - In the present study, we report on the development of bivariate flow karyotyping in the legume broad bean (Vicia faba). We optimised chromosome staining with 4',6 diamidino-2-phenylindole and mithramycin A and analysed chromosome suspensions prepared from a line with standard (wild-type) karyotype and from six translocation lines with reconstructed karyotypes. Chromosomes were isolated from formaldehyde-fixed root tips after cell cycle synchronisation, and their fluorescence was analysed with dual-laser flow cytometry after the staining. High resolution bivariate flow karyotypes were obtained in all broad bean lines analysed. Compared with univariate analysis, the bivariate analysis permitted discrimination of more chromosome types. However, peaks corresponding to newly resolved chromosomes were rather closely spaced, which could have compromised the purity of sorted fractions. With only a few exceptions, chromosome peaks were in a straight line, suggesting only minor differences in the AT:GC ratio among the chromosomes. These results indicate the limited potential of bivariate flow cytometric analysis and sorting in broad bean. PMID- 9222110 TI - Detailed analysis of cell cycle kinetics upon proteasome inhibition. AB - We have studied specific effects of proteasome inhibition on cell cycle progression. To this end, the protease inhibitors MG115, calpain inhibitor I, and calpain inhibitor II, which display differential inhibitory effects on proteasomes, were used. Cell kinetic studies using bromodeoxyuridine pulse labeling revealed a complete block of G1/S and metaphase transitions and a delayed progression through S phase in cell cultures treated with 54 microM of MG115. Calpain inhibitor I in similar concentrations displayed a fivefold lower effect on cell cycle kinetics. Calpain inhibitor II and MG2M, which is a structural analogue of MG115, had no effect on the cell cycle. The inhibitory effect of MG115 treatment was reversible, because the cell cycle was immediately resumed when the MG115-containing culture medium was replaced by fresh culture medium. Because ubiquitinated proteins accumulated after MG115 treatment, it was confirmed that ubiquitin-dependent protein degradation, and thus proteasomal activity were blocked. By comparison of biochemical and in vitro proteasome inhibition experiments, it was hypothesized that chymotrypsin-like activity of proteasomes may play an important role in cell cycle kinetics. PMID- 9222112 TI - An unusual artefact in the terminal deoxynucleotidyl transferase assay for apoptotic cells. AB - A staining artefact associated with the terminal deoxynucleotidyl assay for apoptotic cells is described. the Artefact is caused by particulate matter in the reconstituted dried milk used in the washing buffer. We recommend filtering the washing buffer before use. PMID- 9222111 TI - Correlated flow cytometric analysis of terminal events in apoptosis reveals the absence of some changes in some model systems. AB - Many parallel processes occur during the final stages of apoptosis. It is not clear which of these processes occur in all or most models of apoptosis and which occur only in some. In addition, the temporal relationship of these events is not always well understood. Correlated flow cytometric measurements were used to address these questions. Several models of apoptosis were studied, including thymocytes treated with dexamethasone. MOLT-4 cells treated with etoposide, U937 cells treated with anti-Fas, HL-60 cells treated with camptothecin, Raji cells grown in low serum, and aged neutrophils. All models showed a decrease in LDS-751 and fluorescein diacetate (FDA) staining, an increase in staining with dihydrorhodamine 123 (dhR123) or dihydroethidium, and an acidification of the cytoplasm. In each model, these changes were highly correlated, appearing simultaneously as multiparameter measurements. Changes in membrane status detected with merocyanin 540 (MC540) and annexin V behaved differently. A population with LDS-751 and FDA changes but without annexin V or MC540 changes could be demonstrated in some models. Several models did not show any change in annexin V binding, and HL-60 did not show a change in MC540 binding during apoptosis. The loss of cell surface antigens (CD45 and CD16) from aged neutrophils occurred in the entire LDS-751 and FDA dim population, even though other membrane changes (including the appearance of annexin V binding sites) were only apparent in a subset of these cells. These results suggest a model for the ordering of some of the terminal processes in apoptosis, with annexin V and MC540 changes trailing other events in apoptosis. These results confirm the need for caution in using a single-parameter measurement as an indicator of apoptosis for any new model being studied. PMID- 9222113 TI - Oral grasping of a surrogate nipple by the newborn rat. AB - Newborn rat pups exhibit oral appetitive behaviors when presented with an artificial nipple. These behaviors include mouthing and licking movements and expression of stereotyped oral grasp response. Caesarean-delivered pups show increased responding to the nipple over the first 5 h after birth that is independent of experience with the nipple. Mimicking maternal licking by stimulating the anogenital region of the newborn rat with a soft paintbrush increases response to the nipple. Pups tested after 24 hr of normal experience respond to the artificial nipple when tested immediately after separation from the mother. However, oral grasping of the nipple is more frequent in 1-day-old pups tested 3 or 5 hr after separation from the mother. Study of behavioral responses to the artificial nipple promises to provide information about sensory and neurochemical controls of the initial suckling episode. PMID- 9222114 TI - Offspring-induced nurturance: animal-human parallels. AB - Offspring provide mothers with stimuli that impel their own nurturance. In rats, distal sensory stimuli from pups--sight, sound, odor--contribute to contact seeking, whereas tactile stimuli from pups to dam's snout and ventrum elicit essential maternal behavioral reflexes involved in retrieval, licking, and the quiescent, upright nursing posture (kyphosis). Brain sites involved with maternal behavior--assessed by lesions, immunocytochemical visualization of gene activity, and neurophysiological mapping--include the midbrain central gray, medial preoptic nucleus, limbic system, and somatosensory cortex; these may change with experience. Human mothers inadvertently learn to identify their own baby rapidly after birth and can do so via a single sensory modality. Subsequently maternal responsiveness and gratification are impaired by inappropriate, insufficient, or nonreciprocal interactions such as occurs when the baby cries excessively, is blind, deaf, or autistic. Thus, maternal behavior characterized by elicited responses and emotional reactions to stimuli from offspring may be evolutionarily conserved. PMID- 9222115 TI - Evidence that endogenous corticotropin-releasing factor suppresses behavioral responses of guinea pig pups to brief isolation in novel surroundings. AB - Guinea pig pups were injected subcutaneously with a corticotropin-releasing factor antagonist (CRF12-41) or saline vehicle and then placed into a novel cage for 30 or 60 min. Isolated 20- to 26-day-old pups vocalized more and exhibited more locomotor activity when given 15 to 150 micrograms of CRF12-41 than when given saline. The presence of the mother in the test cage prevented the antagonist from affecting behavior. The influence of the antagonist during isolation was not limited to guinea pigs near weaning age: CRF12-41 increased levels of vocalizing in isolated, 4- to 6-day-old pups, though no changes were seen in locomotor activity. Results support the hypothesis that endogenous corticotropin-releasing factor, perhaps acting at a peripheral binding site, suppresses the active behavioral response characteristic of pups during the early phase of isolation in novel surroundings. PMID- 9222116 TI - Behavioral and physiological characteristics of Indian and Chinese-Indian hybrid rhesus macaque infants. AB - Strain differences in temperament and physiology have been reported for several animal species, but nonhuman primates have not been well studied in this regard. We assessed behavioral and physiology in Chinese-Indian hybrid (n = 13) and Indian-origin (n = 29) nursery-reared rhesus monkey infants. Previous data indicate that Chinese-origin and Chinese-Indian hybrid rhesus exhibit more aggression directed toward humans and conspecifics and are more irritable in response to neonatal assessment procedures than are Indian-derived rhesus. In addition, in rhesus adults, low levels of cerebrospinal fluid 5-HIAA have been correlated with impulsivity, aggressive behavior, and diminished social competence. We therefore hypothesized that hybrid infants would exhibit more behavioral and adrenocortical reactivity in the home cage and during social separations, would be less sociable in their peer groups, and would exhibit lower CSF 5-HIAA levels than Indian-derived monkeys. Blood and cerebrospinal fluid samples were obtained on Days, 14, 30, 60, 90, 120, and 150 of life, and prior to and during social separations at 6 months of age. Behavioral observations were conducted in the home cage and during the separation condition. No differences in behavior were observed between hybrid and Indian-derived animals in the home cage. Indian-derived and hybrid infants exhibited diverging patterns of behavioral reactivity across the 4 weeks of the repetitive social-separation procedure, and during reunion periods. Although plasma cortisol levels were sensitive to the testing conditions, no group differences were observed. CSF 5 HIAA declined over time for all monkeys, and hybrid animals exhibited significantly lower 5-HIAA levels than Indian monkeys beginning at 6 months of age. These findings are consistent with the known behavioral and physiological characteristics of Chinese-origin adult rhesus. PMID- 9222118 TI - Insulin resistance is central to the burden of diabetes. PMID- 9222117 TI - Temperament, social competence, and adrenocortical activity in preschoolers. AB - The relations among temperament, social competence, and levels of a stress sensitive hormone (salivary cortisol) were examined in two studies of preschoolers children (Study 1, N = 29; Study 2, N = 46). In both studies, we sampled cortisol daily for the initial weeks of school year (Group Formation period) and for several weeks later in the year (Familiar Group period). For each child, we examined two measures of cortisol activity (separately for each period) based on the distribution of cortisol levels across days: (a) median cortisol (50th percentile) and (b) cortisol reactivity (the difference between the 75th and 50th percentile). Median cortisol was modestly stable across periods, but cortisol reactivity was not. Children who showed high cortisol reactivity (75th minus 50th percentile > or = 0.10 micrograms/dl) during the Group Formation period but low-to-normal cortisol reactivity during the Familiar Group period were outgoing, competent, and well liked by their peers. In contrast, children who changed from low/normal to high cortisol reactivity and those who maintained high cortisol reactivity from the Group Formation to Familiar Group period were affectively negative and solitary. Children who showed high median cortisol during the Familiar Group period or over both periods scored lower on a measure of attentional and inhibitory control. Together, these results suggest that relations among temperament, social competence, and neuroendocrine reactivity reflect both individual and contextual differences. PMID- 9222119 TI - Insulin resistance is not central to the burden of diabetes. PMID- 9222120 TI - Triglyceride is the major atherogenic lipid in NIDDM. PMID- 9222121 TI - Cholesterol is the major atherogenic lipid in NIDDM. PMID- 9222122 TI - Primary prevention of NIDDM: a practical reality. PMID- 9222123 TI - Primary prevention of NIDDM: a future dream. PMID- 9222124 TI - Reducing the burden: the diabetes debates. PMID- 9222125 TI - Thinking big: four ways to advance environmental health research to answer the needs of public policy. PMID- 9222126 TI - Comparison of 17 methods of predicting the carcinogenicity of 30 chemicals. PMID- 9222127 TI - Purpose and guidelines for toxicokinetic studies within the National Toxicology Program. AB - Toxicokinetic studies undertaken within the National Toxicology Program are intended to aid the design of toxicology and carcinogenicity studies, help interpret the results of toxicology and carcinogenicity studies with respect to the relationship between toxic effects and external exposure, and define the parameters of dose, distribution, metabolism, and elimination that can be used in human risk assessment. Descriptions of two study designs presented here represent the possible extremes in approaches to toxicokinetic studies. The comprehensive approach is geared toward the development of physiology based pharmacokinetic models that relate external exposure to target organ dosimetry and addresses the questions: Is the chemical absorbed? How is the chemical metabolized? Where are the chemical and/or metabolites distributed in the body? What are the elimination rate and route of the chemical? What is the effect of dose on absorption, distribution, metabolism, and elimination? The minimal study design is more limited in scope than the comprehensive design and addresses primarily the issues of absorption, distribution, and elimination of the parent chemical. Study protocols for most chemicals lie somewhere between these two extreme approaches. An increased understanding of the relationships between external exposure, target organ dosimetry, and adverse effects should provide greater confidence in making low-dose extrapolations of human risk. This paper focuses on the collection of data from animal toxicokinetic studies. The construction of comparable models to characterize target organ dosimetry in exposed humans would certainly require the use of human parameter values obtained from human tissue samples and volunteers. PMID- 9222128 TI - Is Bt better? PMID- 9222129 TI - Update on Gulf War illness. PMID- 9222130 TI - Science among the Saguaros. PMID- 9222131 TI - Hormones and health. PMID- 9222132 TI - The laws of genetics. PMID- 9222133 TI - Mobile environmental labs. PMID- 9222134 TI - A lung retention model based on Michaelis-Menten-like kinetics. AB - A Michaelis-Menten (MM)-like kinetic model for pulmonary clearance and retention of insoluble dusts was developed and validated by comparing our predictions with experimental data from F344 rats. Published data from inhalation studies involving accumulation and elimination of photocopy test toner, antimony trioxide, carbon black, and diesel exhaust particles were investigated. Numerical integration techniques were used to solve mass balance relationships based upon dust retention in a single lung compartment and clearance via an MM-like kinetic process. The model fit most of the experimental data well. The parameters of MM like clearance kinetics, which had been derived strictly from the elimination phase, accurately predicted dust retention during the elimination as well as accumulation phases. Furthermore, parameters estimated from one study could accurately predict retention of the same dust in other studies. Particle density and gender of the animals had no effect on the goodness of fit of model predictions. This study suggests that MM-like kinetics offer a reasonable description of particle clearance from the pulmonary region of the rat lung that is more parsimonious than existing particle-clearance models and therefore more suitable for use with small amounts of data. PMID- 9222135 TI - Talc and amosite/crocidolite preferentially deposited in the lungs of nonoccupational female lung cancer cases in urban areas of Japan. AB - To analyze the correlation between asbestos lung burden and lung cancer, lungs of 211 female cases with and without lung cancer were examined. Phase-contrast microscopic (PCM) counting of ferruginous (FBs) and uncoated fibers (UFs), which had length longer than 5 microns and aspect ratios greater than 3:1, revealed a significantly higher level of FBs plus UFs in urban lung cancer cases than urban non-lung cancer cases (1380.5 vs. 550.3; p < 0.001). No difference was noted between rural lung cancer and non-lung cancer cases. Analytical electron microscopic studies identified various kinds of mineral fibers with an aspect ratio greater than 3:1 in the lung tissue including chrysotile, actinolite/tremolite, amosite/crocidolite, fibrous talc, mica, silica, iron, wollastonite, chlorite, kaoline, and others. The most frequently detected fibers were thin, short chrysotile fibers, most of which could not be found by PCM, followed by relatively thick, long actinolite/tremolite fibers, fibrous talc, and in a smaller number, amosite/crocidolite of intermediate length and width. Amosite/crocidolite and fibrous talc counts in urban lung cancer cases were greater than those of urban non-lung cancer cases, rural lung cancer, and rural non-lung cancer cases; these findings were consistent with PCM analysis. Therefore, it is suggested that fibers detected in PCM observation may be mainly amosite/crocidolite with some parts fibrous talc and that fibrous talc in urban environments may be another candidate for carcinogenic or cocarcinogenic factors of female lung cancer. PMID- 9222136 TI - Correlation of environmental carbaryl measurements with serum and urinary 1 naphthol measurements in a farmer applicator and his family. AB - In exposure or risk assessments, both environmental and biological measurements are often used. Environmental measurements are an excellent means for evaluating regulatory compliance, but the models used to estimate body burden from these measurements are complex. Unless all possible routes of exposure (i.e., inhalation, dermal absorption, ingestion) are evaluated, exposure to a toxicant can be underestimated. To circumvent this problem, measurements of the internal dose of a toxicant in blood, serum, urine, or tissues can be used singularly or in combination with environmental data for exposure assessment. In three separate laboratories, carbaryl or its primary metabolite, 1-naphthol, was measured in personal air, dermal samples, blood serum, and urine from farmer applicators and their families. The usefulness of both environmental and biological data has been demonstrated. For the farmer applicator, the environmental levels of carbaryl would have been sufficient to determine that an exposure had occurred. However, biological measurements were necessary to determine the absorbed dose of each member of the applicator's family. In addition, a correlation between serum and urinary 1-naphthol measurements has been shown; therefore, either matrix can be used to accurately evaluate occupational carbaryl exposure. PMID- 9222137 TI - Association between air pollution and low birth weight: a community-based study. AB - The relationship between maternal exposure to air pollution during periods of pregnancy (entire and specific periods) and birth weight was investigated in a well-defined cohort. Between 1988 and 1991, all pregnant women living in four residential areas of Beijing were registered and followed from early pregnancy until delivery. Information on individual mothers and infants was collected. Daily air pollution data were obtained independently. The sample for analysis included 74,671 first-parity live births were gestational age 37-44 weeks. Multiple linear regression and logistic regression were used to estimate the effects of air pollution on birth weight and low birth weight (< 2,500 g), adjusting for gestational age, residence, year of birth, maternal age, and infant gender. There was a significant exposure-response relationship between maternal exposures to sulfur dioxide (SO2) and total suspended particles (TSP) during the third trimester of pregnancy and infant birth weight. The adjusted odds ratio for low birth weight was 1.11 (95% CI, 1.06-1.16) for each 100 micrograms/m3 increase in SO2 and 1.10 (95% CI, 1.05-1.14) for each 100 micrograms/m3 increase in TSP. The estimated reduction in birth weight was 7.3 g and 6.9 g for each 100 micrograms/m3 increase in SO2 and in TSP, respectively. The birth weight distribution of the high-exposure group was more skewed toward the left tail (i.e., with higher proportion of births < 2,500 g) than that of the low-exposure group. Although the effects of other unmeasured risk factors cannot be excluded with certainty, our data suggests that TSP and SO2, or a more complex pollution mixture associated with these pollutants, contribute to an excess risk of low birth weight in the Beijing population. PMID- 9222138 TI - High lead exposure and auditory sensory-neural function in Andean children. AB - We investigated blood lead (B-Pb) and mercury (B-Hg) levels and auditory sensory neural function in 62 Andean school children living in a Pb-contaminated area of Ecuador and 14 children in a neighboring gold mining area with no known Pb exposure. The median B-Pb level for 62 children in the Pb-exposed group was 52.6 micrograms/dl (range 9.9-110.0 micrograms/dl) compared with 6.4 micrograms/dl (range 3.9-12.0 micrograms/dl) for the children in the non-Pb exposed group; the differences were statistically significant (p < 0.001). Auditory thresholds for the Pb-exposed group were normal at the pure tone frequencies of 0.25-8 kHz over the entire range of B-Pb levels, Auditory brain stem response tests in seven children with high B-Pb levels showed normal absolute peak and interpeak latencies. The median B-Hg levels were 0.16 micrograms/dl (range 0.04-0.58 micrograms/dl) for children in the Pb-exposed group and 0.22 micrograms/dl (range 0.1-0.44 micrograms/dl) for children in the non-Pb exposed gold mining area, and showed no significant relationship to auditory function. PMID- 9222140 TI - Asthma and the environment. PMID- 9222142 TI - On the natural language of signs; and its value and uses in the instruction of the deaf and dumb. 1848. PMID- 9222139 TI - Alterations in steroidogenesis in alligators (Alligator mississippiensis) exposed naturally and experimentally to environmental contaminants. AB - Many environmental contaminants alter the reproduction of animals by altering the development and function of the endocrine system. The ability of environmental contaminants to alter the endocrine system of alligators was studied both in a descriptive study in which juvenile alligators from a historically contaminated lake were compared to animals from a control lake and in an experimental study in which hatchling control alligators were exposed in ovo to several endocrine disrupting standards and two modern-use herbicides. Endocrine status was assessed by examining plasma hormone concentrations, gonadal-adrenal mesonephros (GAM) aromatase activity, and gonadal histopathology. In the descriptive study, juvenile alligators from the contaminated lake had significantly lower plasma testosterone concentrations (29.2 pg/ml compared to 51.3 pg/ml), whereas plasma 17 beta-estradiol concentrations did not vary when compared to controls. GAM aromatase activity was significantly decreased n the alligators from the contaminated lake (7.6 pmol/g/hr compared to 11.4 pmol/g/hr). In the experimental study, the endocrine-disrupting standards had the expected effects. 17 beta Estradiol and tamoxifen caused sex reversal from male to female, with a corresponding increase in aromatase activity. Vinclozolin had no apparent effect on male or female alligators. Among the herbicides tested, atrazine induced GAM aromatase activity in male hatchling alligators that was neither characteristic of males nor females, although testicular differentiation was not altered. Exposure to 2,4-dichlorophenoxyacetic acid had no effect on the endocrine parameters that were measured. Together, these studies show that exposure to some environmental chemicals (such as atrazine) can alter steroidogenesis in alligators, but the endocrine alterations previously noted for Lake Apopka, Florida, alligators can not be fully explained by this mechanism. PMID- 9222143 TI - Notions of the deaf and dumb before instruction, especially in regard to religious subjects. 1855. PMID- 9222144 TI - Words recognized as units; systematic signs. 1859. PMID- 9222145 TI - Is the sign-language used to excess in teaching deaf-mutes? 1871. PMID- 9222146 TI - Reflections of a deaf-mute before education. Narration by Mr. Ballard. 1881. PMID- 9222147 TI - The value of the sign-language to the deaf. 1887. PMID- 9222148 TI - Must the sign-language go? 1899. PMID- 9222149 TI - An investigation concerning the value of the oral method. 1909. PMID- 9222150 TI - Mistaken investigations concerning the value of the oral method. 1910. PMID- 9222151 TI - Americanization in our schools for the deaf. 1921. PMID- 9222152 TI - The status of the preschool deaf child. 1934. PMID- 9222153 TI - Professional preparation and advancement of deaf teachers. 1941. PMID- 9222155 TI - The influence of early manual communication on the linguistic development of deaf children. 1966. PMID- 9222154 TI - The educational preparation of oral teachers of the deaf. 1952. PMID- 9222156 TI - Psycholinguistics and deafness. 1970. PMID- 9222157 TI - Signs and manual communication systems: selection, standardization, and development. 1978. PMID- 9222158 TI - Bi-Bi to MCE? 1993. PMID- 9222159 TI - The decline of Haemophilus influenzae type b disease in Australia. AB - Between July 1993 and June 1996, there were 412 cases of invasive Haemophilus influenzae type b (Hib) disease reported to the Hib Case Surveillance Scheme, 71% in children under the age of five years. Meningitis was the most frequent illness reported, followed by epiglottitis, septicaemia and pneumonia. There were 18 deaths. Thirty-four cases were classified as vaccine failures. The number of vaccine failures increased over time and the total number of cases of Hib disease fell, consistent with an increase in Hib vaccine coverage. Based on an estimated vaccine coverage of 50% in April 1995, the vaccine efficacy for all vaccines in the period was estimated to be 89%. Invasive Hib is a serious illness of childhood which is being significantly reduced by the use of Hib vaccines, and has the potential to be eliminated from this country. Vaccination providers should aim to immunise all children against Hib disease on time and according to the National Health and Medical Research Council Standard Vaccination Schedule. PMID- 9222160 TI - Communicable diseases surveillance. PMID- 9222162 TI - Policies on reporting clinical trials and publishing supplements. PMID- 9222161 TI - Changes in policy for publishing manuscripts in Neurology. PMID- 9222163 TI - Attitudes of US neurologists concerning the ethical dimensions of managed care. AB - We surveyed attitudes of US neurologists about the ethical dimensions of managed care by administering a written instrument containing paradigmatic cases portraying conflicts of physicians, patients, and managed care organizations (MCOs). After each case, we assessed neurologists' attitudes by asking them their degree of agreement with a series of statements. We found that neurologists (1) generally were willing to follow clinical practice guidelines if they were created by medical societies; (2) experienced frequent conflicts of interest or conflicts of obligation in the care of their MCO patients; (3) feared legal ramifications of their clinical decisions on MCO patients; (4) were unwilling to employ deception or gaming to achieve what they perceived to be good patient care; (5) believed that their professional prerogatives and autonomy were under attack by MCOs; and (6) felt that the good of their patients should not be sacrificed for the good of society. PMID- 9222164 TI - Intravenous thrombolysis for acute stroke. PMID- 9222165 TI - Double-crush syndrome: a critical analysis. PMID- 9222166 TI - An association between head circumference and Alzheimer's disease in a population based study of aging and dementia. AB - We investigated the association between head circumference (HC) and Alzheimer's disease (AD) in a cross-sectional population-based study of aging in North Manhattan. Six hundred forty-nine subjects underwent neurologic, neuropsychological, and anthropometric evaluations; apolipoprotein E (apoE) genotype was available for a subsample of 300 individuals. Logistic regression analyses were performed with AD the outcome of interest to evaluate any association between HC and AD. In these analyses, HC evaluated as a continuous variable was associated with AD (OR 0.8, 95% CI 0.7-0.9) after adjusting for age, education, and ethnicity, gender, and height. Analyses suggested that increased risk resided mainly in those with smallest HC. Thus, women whose HC was within the lowest quintile of HC for women were 2.9 (95% CI 1.4-6.1) times more likely to have AD, after adjusting for age, education, and ethnicity; and men in the lowest quintile of HC (for men) were 2.3 times more likely to have AD (95% CI 0.6 9.8). There was no confounding by height, weight, or apoE genotype. The results are consistent with previous studies that suggest that premorbid brain size may influence the age-specific risk for AD. Future epidemiologic studies seeking environmental risk factors for AD may benefit by making HC measurements on all subjects to decrease the variance associated with other potential risk factors. PMID- 9222167 TI - Participation in clinical trials and long-term outcomes in Alzheimer's disease. AB - The objective of this study was to determine whether participation in clinical trials affects long-term outcomes in Alzheimer's disease (AD). Participation in clinical trials for persons with dementia is often justified on the grounds that patients benefit from the medical oversight typical of trials, even when experimental agents do not demonstrate short-term benefits. This claim has not been rigorously assessed. Of 215 community-resident subjects enrolled in a prospective study of outcomes in AD, 101 participated in randomized clinical trials (RCTs) during the first 2 years of follow-up. These subjects were compared with subjects who met eligibility requirements for RCTs but did not participate (N = 57) and with subjects who were ineligible (N = 57), over a total of 3.5 years of follow-up. Survival analyses assessed risk of death, nursing home placement, and incident functional deficit end points, adjusting for baseline differences. Subjects who participated in RCTs were younger and more highly educated. Mortality, risk of hospitalization, number of medical examinations conducted by study physicians, and onset of severe functional deficit did not differ between the groups, but risk of nursing home admission was significantly lower among RCT participants compared with eligible nonparticipants and ineligible subjects (16.8% versus 36.8% and 31.6%, respectively [p = 0.01]). The difference in risk of nursing home placement may represent a long-term, drug related benefit to patients, a selection effect (caregivers of patients who participate in RCTs differ from caregivers of patients who do not), or a positive effect on caregivers (greater contact with a medical service may be associated with better care-giving outcomes). Further research is required to assess these effects. PMID- 9222168 TI - Incidence and prevalence of dementia in a multiethnic cohort of municipal retirees. AB - BACKGROUND: Multiethnic population-based studies of dementia are lacking in the literature; therefore, we conducted a study to determine the incidence and prevalence of dementia among, black, white, and Hispanic municipal retirees age 50 and older. METHODS: In 1991, city of Houston municipal workers who retired between 1980 and 1984 received in-home screening for cognitive impairment using the Mini-Mental State Examination and were referred for comprehensive neurologic evaluation if indicated. Families of deceased retirees were interviewed, medical records were reviewed, and death certificates were obtained to determine case status of the retiree. Study participation was 90%. RESULTS: Crude prevalence of dementia, among retirees age 60 and older, was 2.65 per 100 population. Age adjusted to the 1970 US population, age 60 to 80, the prevalence was 1.85 per 100 population. Age-adjusted prevalence of dementia was similar among Hispanic and black men, 4.75 and 4.80 respectively, and lowest among white men, 2.42 per 100 population. Forty-three percent of the prevalent cases (30% of the incidence cases) were not previously diagnosed. The cumulative incidence of dementia calculated to age 80 was 39% for Hispanic men, 28% for black men, and 14% for white men. Fifty-five percent of the incidence cases were diagnosed as having ischemic vascular dementia (IVD), 20% as having probable or possible Alzheimer's dementia (AD), and 25% as having unspecified dementia. CONCLUSIONS: Risk of dementia by age 80 was more pronounced for Hispanic and black men compared with white men. IVD was the predominant cause of dementia among both black and white men. Grouping Hispanics with whites may mask differences when studying dementia. PMID- 9222169 TI - Apolipoprotein E genotypes in primary progressive aphasia. AB - We obtained apolipoprotein E genotyping in a population of 12 consecutive patients who fulfilled rigorous criteria for the clinical diagnosis of primary progressive aphasia (PPA). The allele frequencies were 4% for E2, 83% for E3, and 13% for E4. This pattern of allele distribution was significantly different from the pattern seen in groups of patients either with the clinical diagnosis of probable Alzheimer's disease (PRAD) or the histopathologic diagnosis of Alzheimer's disease (AD). The E4 allele frequency in the group of patients with PPA was in the range seen in control populations and was much lower than the one reported in populations of patients with PRAD or AD. The E4 allele is therefore not a significant risk factor for developing PPA. These results provide neurobiological support for the syndromic distinction of PPA from PRAD and are in keeping with neuropathologic evidence showing that the vast majority of patients with PPA do not have the histopathology of AD. Although we do not yet have neuropathologic information on our patients, these results indicate that the clinical diagnosis of PPA has biological validity in that it identifies a population that is genetically different from the population of patients with a clinical diagnosis of PRAD. PMID- 9222170 TI - Genetic association of the low-density lipoprotein receptor-related protein gene (LRP), an apolipoprotein E receptor, with late-onset Alzheimer's disease. AB - The presence of the APOE epsilon 4 allele encoding apolipoprotein E4 (apoE4) is the major genetic risk factor for late-onset Alzheimer's disease (AD). However, the molecular and cellular mechanisms by which APOE epsilon 4 renders AD risk are unclear. In this report, we present genetic evidence that an apoE receptor, LRP, may be associated with the expression of late-onset AD. Using a biallelic genetic marker in exon 3 of LRP, late-onset AD cases markedly differed from the control subjects in the distribution of LRP genotypes, and this difference was highly accentuated among AD cases with positive family history of senile dementia. Furthermore, the numbers of neutritic plaques were significantly altered as a consequence of different LRP genotypes in postmortem AD cases. Taken together, our results implicate the pathophysiology of LRP in the expression of late-onset AD. PMID- 9222171 TI - What are the obstacles for an accurate clinical diagnosis of Pick's disease? A clinicopathologic study. AB - Several studies have evaluated the reliability and validity of the clinical diagnosis of Alzheimer's disease (AD) using well-defined neuropathologic criteria, but none has attempted to evaluate the diagnostic accuracy of Pick's disease. We determined the accuracy of the clinical diagnosis of Pick's by presenting 105 autopsy-confirmed cases of Pick's (n = 7) and related disorders (non-Pick's, n = 98) as clinical vignettes in randomized order to six neurologists who were unaware of the autopsy findings. The group of raters had moderate to fair agreement for the diagnosis of Pick's as measured by the kappa statistics. The sensitivity for the diagnosis of Pick's for the first visit (mean, 53 months after onset) and last visit (mean, 78 months after onset) was low (range, 0 to 71%), but specificity was near-perfect. Median positive predictive values at both visits were 83 to 85%. False-negative misdiagnoses mainly involved AD. False-positive diagnoses were rare and occurred with corticobasal degeneration (first visit) and with dementia with Lewy bodies (last visit). Pick's was also misdiagnosed by primary neurologists. The best clinical predictors for the early diagnosis of Pick's included "frontal" dementia, early "cortical" dementia with severe frontal lobe disturbances, absence of apraxia, and absence of gait disturbance at onset. However, the first neurologic evaluation in some of the Pick's cases took place in advanced stages of the disease. Our findings suggest that this disorder is underdiagnosed in clinical practice. Although the low sensitivity for the clinical diagnosis of Pick's is disappointing, our data suggest that when clinicians suspect Pick's, their diagnosis is almost always correct. Absence of awareness of the main features of this disorder and of specificity of the frontal lobe syndrome may partially explain the low detection of Pick's disease. PMID- 9222172 TI - The relationship between extrapyramidal signs and cognitive performance in patients with Alzheimer's disease enrolled in the CERAD Study. Consortium to Establish a Registry for Alzheimer's Disease. AB - The objective of this study was to determine the relationship between the presence of extrapyramidal signs and the severity of cognitive and functional impairment in patients with Alzheimer's disease (AD). Eleven university medical centers in the United States and France participated in the Consortium to Establish a Registry for Alzheimer's Disease (CERAD) study of extrapyramidal signs in AD. Forty-seven patients with probable AD who had extrapyramidal signs were matched by sex, race, education, and age with 132 probable AD patients without extrapyramidal signs. The main outcome measures were the Clinical Dementia Rating, Blessed Dementia Scale, and the CERAD Neuropsychology Battery (verbal fluency, naming, Mini-Mental State Examination, word list learning, word list recall, savings ratio, word list recognition, and constructional praxis). AD patients with extrapyramidal signs performed more poorly than AD patients without parkinsonism on various neuropsychological tests, even after controlling for the Clinical Dementia Rating and reported duration of cognitive impairment. The severity of the extrapyramidal manifestations was related to the degree of cognitive and functional impairment. Muscular rigidity and bradykinesia were the most frequent extrapyramidal signs associated with AD. Patients with AD associated with extrapyramidal signs have greater cognitive and functional impairment than AD patients without clinical evidence of parkinsonism. PMID- 9222173 TI - The impact of apolipoprotein E4 on cause of death in Alzheimer's disease. AB - OBJECTIVE: We tested the hypothesis that the apolipoprotein E epsilon 4 (apoE4) allele is associated with an increased proportion of vascular-related mortality in Alzheimer's disease (AD). BACKGROUND: ApoE4 is associated with an increased risk of developing AD, with an earlier onset, and may predispose to vascular dementia as well. In the general population, apoE4 has been associated with increased coronary artery disease and shorter lifespan. There is a paucity of data regarding the effect of the apolipoprotein E (apoE) genotype upon the contributing causes of death in AD. METHODS: Death certificates of 114 AD cases were reviewed blind to apoE genotype. Deaths due to ischemic heart disease (IHD), cerebrovascular disease (CVD), vascular disease (either IHD or CVD), pneumonia, and other causes were analyzed as a function of apoE genotype. Logistic regression analyses were employed to control for age and gender effects. RESULTS: The likelihood of vascular disease contributing to death increased in association with the epsilon 4 allele (29% in cases without an epsilon 4 allele, 43% in cases with one epsilon 4 allele, 53% in epsilon 4/4 homozygous cases; p = 0.035 after corrections for age and gender). This increase appeared largely due to an increase in ischemic heart disease, which was reported more frequently on death certificates of cases with one or more epsilon 4 allele (adjusted odds ratio [OR] = 1.85 per epsilon 4 allele; p < 0.05). There were nonsignificant trends for apoE4 to be associated with increased mortality related to cerebrovascular disease (OR = 1.45) and decreased mortality related to pneumonia (OR = 0.77) and AD itself (OR = 0.72). The epsilon 4/4 cases had significantly earlier age of onset (mean = 64.5 yr), earlier death, and longer duration of disease (mean = 10.1 yr). Cases with one or more epsilon 4 allele tended to have lower mean MMSE scores prior to death (6.6 versus 9.5) and were more often female (54% versus 45%). CONCLUSIONS: The apoE4 allele appears to increase the risk of vascular and ischemic heart disease-related death in patients with AD. PMID- 9222174 TI - Signed and spoken language perception studied by positron emission tomography. AB - Sign and spoken language seem to be localized in the same brain areas. They elicit similar regional cerebral blood flow (rCBF) patterns, even though sign language is dependent on spatial information. We investigated sign and spoken language perception in a group of healthy bilingual subjects. Four videotaped activation conditions were used during PET imaging: (1) sign language, (2) spoken language, (3) spoken language with mouth covered, and (4) spoken language on a sound track while showing a motionless face. Spoken language (condition 4) activated significantly the perisylvian cortex (Brodmann areas 22 and 43) bilaterally. Sign language activated the visual association areas (Brodmann areas 37 and 19) but did not selectively activate parietal regions. A reciprocal relationship was observed between the level of activation in visual language perception areas and that in auditory perception areas. We conclude that when healthy bilingual subjects use the visual route for sign language perception, the functional anatomy overlaps that of language processing containing both auditory and visual components. PMID- 9222175 TI - Hemianopic anosognosia. AB - In a prospective study of 32 consecutive patients with homonymous visual field defects due to ischemic infarcts we found hemianopic anosognosia (HAN), defined as the unawareness of visual loss in the homonymous hemifield (or hemiquadrant), in 20 patients (62%). HAN, although occurring predominantly in right-side lesions in 16 of 26 patients (62%) was also present in four of six patients (or 67%) with left-side lesions. This group of patients has been presented in a prior report on positive spontaneous visual phenomena. HAN was associated with somatic anosognosia in nine patients and hemineglect in 17 patients. Dissociation between somatic and hemianopic anosognosia, as well as between hemineglect and HAN, was present in several patients, indicating that these phenomena may be independent of each other. Eight patients had pure homonymous hemianopia; that is, hemianopia without cognitive, motor, or somatosensory deficits. Four of these patients (Group A) had awareness of the visual deficit, whereas three patients (Group B) had HAN. Patients in these two groups had similar anatomic lesions. Patients with phosphenes, photopsias, or visual hallucinations were usually aware of their visual field loss. We suggest that HAN is most often related to failure of discovery of the deficits, occasionally to severe visual hemineglect, sometimes to generalized cognitive impairment, or to a combination of these factors. We further conclude (1) there is no specific cortical area for conscious visual perception; (2) visual awareness is processed by a distributed network including multiple visual cortices, parietal and frontal lobes, the pulvinar, and lateral geniculate bodies (lesions localized at various nodes or centers in the network may produce similar phenomena); and (3) both hemispheres are involved in visual processing and conscious awareness. PMID- 9222176 TI - Effects of new anticonvulsant medications on porphyrin synthesis in cultured liver cells: potential implications for patients with acute porphyria. AB - Some patients with acute hereditary porphyrias have seizures and require anticonvulsant therapy, but many anticonvulsants induce exacerbations of the hepatic porphyrias. Recently, several new anticonvulsants have become available. Among these are gabapentin, vigabatrin, felbamate, lamotrigine, and tiagabine. Little is known about their potential for induction of porphyric attacks. We used a cell culture model of primary chicken embryo liver cells, which maintain intact heme synthesis and regulation, to study the effects of these new anticonvulsants on porphyrin accumulation. Treatment of the cells with deferoxamine (250 microM) led to a partial block in heme synthesis, simulating the conditions encountered in human beings with porphyria. Concomitant exposure of these cells to phenobarbital (2 mM) strongly induced accumulation of porphyrins, serving as a positive control in this model. Cells were treated for 20 hours with increasing doses (3.2 to 1,000 microM) of the newer anticonvulsants, with or without deferoxamine. For most of these anticonvulsants 5 to 100 microM is representative of the concentrations achieved in humans with therapeutic doses. Porphyrins were measured spectrofluorometrically as uro-, copro-, and protoporphyrins. Results were confirmed by high-pressure liquid chromatography. Neither vigabatrin nor gabapentin treatment, with or without deferoxamine, led to any increase in porphyrin accumulation. Similar doses of felbamate (with deferoxamine) led to a marked increase in (mainly proto-) porphyrin levels, qualitatively and quantitatively almost identical to the accumulation produced by phenobarbital. Lamotrigine or tiagabine (with deferoxamine) caused similar porphyrin accumulation. Tiagabine treatment up to 100 microM (with deferoxamine) also resulted in very high levels of predominantly proto-porphyrin. In contrast to the other anticonvulsants tested, tiagabine without deferoxamine led to mild porphyrin accumulation. In the presence of deferoxamine, phenobarbital, felbamate, lamotrigine, or tiagabine, but not gabapentin or vigabatrin, increased levels of the mRNA of ALA synthase, the first and rate-controlling enzyme of porphyrin synthesis. Such enzyme induction is a sine qua non for acute porphyric attacks. We conclude that neither vigabatrin nor gabapentin is porphyrogenic, whereas felbamate, lamotrigine, and, especially, tiagabine lead to much accumulation of porphyrins. The latter three anticonvulsants, therefore, may precipitate or exacerbate acute porphyric attacks in humans. We recommend use of vigabatrin or gabapentin, but not felbamate, lamotrigine, or tiagabine, in patients with acute porphyria and seizures. PMID- 9222177 TI - Recurrent intracerebral hemorrhage. AB - Between 1984 and 1994, of the 375 patients admitted to our department for intracerebral hemorrhage (ICH), 24 (6.4%) had a recurrent ICH. There were 15 women and nine men and the mean age of the patients was 64.7 +/- 9.4 years (range 49-81) at the first bleeding episode and 68.7 +/- 7.5 years (range 57-83) at the second. The mean interval between the two bleeding episodes was 47.5 +/- 30.5 months (range 3 months to 14.8 years). Nine patients presented with more than one recurrence of ICH. Seventy-one percent of the patients were hypertensive. The site of the first hemorrhage was lobar in 17 patients, ganglionic (putamen, thalamus, or caudate nucleus) in six patients, and subdural in one. The recurrent hemorrhage occurred at a different location from the previous ICH. The most common pattern of recurrence was "lobar-lobar" (14 patients) and more rarely "ganglionic-ganglionic" (five patients), which was always observed in hypertensive patients. The outcome after the recurrent hemorrhage was usually poor, with severe cognitive impairment. By comparison with 81 patients followed up to 24 months (47.9 +/- 22.2 months) with isolated ICH without recurrence, only lobar hematoma and a younger age were risk factors for recurrences whereas sex and previous hypertension were not. The mechanisms of recurrence of ICH were multiple (hypertension, cerebral amyloid angiopathy). Control of blood pressure after the first hemorrhage may prevent ICH recurrences. PMID- 9222178 TI - Time course of the apparent diffusion coefficient (ADC) abnormality in human stroke. AB - Diffusion-weighted MRI can rapidly detect acute cerebral ischemic injury as hyperintense signal changes, reflecting a decline in the apparent diffusion coefficient (ADC) of water through brain parenchyma, whereas ADC is elevated in the chronic stage because of increased extracellular water content. To determine the time course of these ADC changes, we analyzed 157 diffusion-weighted MRI studies performed at varying time points from the initial ischemic event from 101 patients. Data were expressed as the relative ADC (rADC), the ratio of lesion to control regions of interest. We observed two phases in the time course of rADC changes in acute human stroke: a significant (p < 0.005) reduction in rADC lasting for at least 96 hours from stroke onset (mean, 58.3% of control; SEM, 1.47) and an increasing trend from reduction to pseudonormalization to elevation of rADC values at later subacute to chronic time points (> or = 7 days). We suggest that the persistent reduction of rADC within the first four days may reflect ongoing or progressive cytotoxic edema to a greater degree than extracellular edema and cell lysis. PMID- 9222179 TI - Central poststroke pain and Wallenberg's lateral medullary infarction: frequency, character, and determinants in 63 patients. AB - Central poststroke pain (CPSP) is an infrequently recognized complication of lateral medullary infarction (LMI). We determined the frequency, nature, and predictors of this complication in 63 patients with LMI. The hypothesis tested was that the degree of clinical sensory loss and extent of infarction seen on MRI, both graded by a predetermined scoring scale, would be predictive of CPSP. We also performed quantitative sensory testing (QST) of thermal and pressure sensation thresholds in a subgroup of 19 patients (nine with CPSP and 10 without) to analyze in detail the spinothalamic and trigeminothalamic systems mediating these modalities from both sides of the face and body. We analyzed these results for specific markers of CPSP. RESULTS: CPSP developed in 25% (16/63) of the patients, all within 6 months. This was constant and severe with frequent allodynia, but responded in all cases to amitriptyline and recurred promptly on attempted weaning. CPSP affected the ipsilateral peri-orbital region most commonly, either alone or in combination with the contralateral limbs. Ipsilateral neurotrophic facial ulceration developed in two cases. CPSP correlated significantly (Fisher's exact test, p < 0.0002) with the degree of clinical sensory loss but not with the size of infarction seen on MRI (Fisher's exact test, p = 0.7). QST revealed a highly specific (100%) and sensitive (89%) finding for CPSP-thresholds from the check contralateral to the LMI were normal in eight of nine cases with CPSP and abnormal in all of the 10 cases without CPSP. Abnormalities in the face contralateral to the infarct are referable to the crossed trigeminothalamic tract in the medullary reticular formation medial to the infarcted lateral medulla. We conclude that this argues for the theory that central pain is caused by denervation sensitivity of the "paleo"-reticulothalamic connections due to a selective "neo"-spinothalamic lesion. PMID- 9222180 TI - Cerebral metabolism in fatal familial insomnia: relation to duration, neuropathology, and distribution of protease-resistant prion protein. AB - We used [18F]-2-fluoro-2-deoxy-D-glucose (FDG) and PET to study regional cerebral glucose utilization in seven patients with fatal familial insomnia (FFI), an inherited prion disease with a mutation at codon 178 of the prion protein gene. Four patients were methionine/methionine homozygotes at codon 129 (symptom duration, 8.5 +/- 1 months) and three were methionine/valine (MET/VAL129) heterozygotes (symptom duration, 35 +/- 11 months). A severely reduced glucose utilization of the thalamus and a mild hypometabolism of the cingulate cortex were found in all FFI patients. In six subjects the brain hypometabolism also affected the basal and lateral frontal cortex, the caudate nucleus, and the middle and inferior temporal cortex. Comparison between homozygous or heterozygous patients at codon 129 showed that the hypometabolism was more widespread in the MET/VAL129 group, which had a significantly longer symptom duration at the time of [18F] FDG PET study. Comparison between neuropathologic and [18F] FDG PET findings in six patients showed that areas with neuronal loss were also hypometabolic. However, cerebral hypometabolism was more widespread than the histopathologic changes and significantly correlated with the presence of protease-resistant prion protein (PrPres). Our findings indicate that hypometabolism of the thalamus and cingulate cortex is the hallmark of FFI, while the involvement of other brain regions depends on the duration of symptoms and some unknown factors specific to each patient. The present data also support the notion that PrPres formation is the cause of neuronal dysfunction in prion diseases. PMID- 9222181 TI - Familial prion disease with a novel 144-bp insertion in the prion protein gene in a Basque family. AB - Three members of a Basque family carrying a novel six R2 octapeptide repeat 144 bp insertion in the prion protein gene (PRNP) showed a slowly progressive dementia associated with cerebellar signs, myoclonic jerks, and seizures. Although postmortem examination revealed only focal and minimal spongiform degeneration in one subject with a 4-year course, significant astrogliosis and neuronal loss were associated with pronounced spongiform degeneration in the patient with a duration of symptoms of 10 years. Prion protein (PrP) immunoreactive patches with a unique morphology were present in the molecular layer of the cerebellum in both subjects. Western blot analysis demonstrated the presence of protease-resistant prion protein (PrPres) with the same characteristics (size and ratio of the three differently glycosylated isoforms) of that found in typical sporadic Creutzfeldt-Jakob disease (CJD129M/M, PrPres type 1). The amount of PrPres correlated with presence and severity of spongiform degeneration in the cerebral cortex. The findings suggest that a relatively low rate of PrPres deposition is the cause of the lack of spongiform degeneration in subjects carrying a 144-bp insertion in PRNP. The presence of PrP-immunoreactive patches with unique morphology in the molecular layer of the cerebellum is a hallmark of certain prion encephalopathies with insertional mutations and is useful in the diagnosis of this subtype of human prion disease. PMID- 9222182 TI - Safety and tolerability of the antioxidant OPC-14117 in HIV-associated cognitive impairment. The Dana Consortium on the Therapy of HIV Dementia and Related Cognitive Disorders. AB - Cognitive impairment is a common and disabling complication of advanced HIV infection. Antiretroviral agents are the only proven therapies currently used for the treatment of HIV dementia, but the response to these agents is frequently unsatisfactory, short-lived, or complicated by intolerable side effects. We hypothesized that OPC-14117, a lipophilic antioxidant that acts to scavenge superoxide anion radicals, might ameliorate the toxic interactions between HIV infected macrophages and neurons. We conducted a double-blind, placebo controlled, randomized clinical trial to assess the safety and tolerability of OPC-14117 240 mg per day. All 30 patients enrolled (15 per group) had cognitive impairment based on performance on neuropsychological tests. The primary outcome was tolerability of the study drug as measured by the proportion of subjects able to complete the study on their assigned dosage of experimental medication. Overall OPC-14117 was as well tolerated as placebo. Five subjects withdrew because of adverse experiences (two placebo, three OPC-14117). The OPC-14117 treated group had better scores on a clinical global impression scale, compared with the placebo group. There were trends toward improvement in the cognitive test scores; however, these changes were not statistically significant. These results demonstrate that this antioxidant intervention is well tolerated in cognitively impaired patients with advanced HIV infection, and suggest that a larger efficacy trial to assess the impact of OPC-14117 on cognitive performance is warranted. PMID- 9222183 TI - Interleukin-6 is expressed at high levels in the CNS in Lyme neuroborreliosis. AB - In patients with Lyme neuroborreliosis, inflammation and symptoms of fatigue and malaise occur out of proportion to the relatively low number of spirochetes present. Previous studies have identified interleukin-6 (IL-6) as a candidate molecule for amplification of CNS inflammation in this disease. We pursued this possibility by measuring cytokine gene expression by reverse-transcriptase polymerase chain reaction (RT-PCR) in the brain of rhesus macaques actively infected with Borrelia burgdorferi. Samples of brain tissue were screened for IL 6 and interferon gamma using RT-PCR-ELISA, a technique that uses RT-PCR, subsequent hybridization of the PCR product with a biotinylated probe, and capture and ELISA readout of hybridization product. The number of copies in positive samples was then quantitated using qRT-PCR-ELISA, in which wild-type cytokine cDNA competes with recombinant competitor DNA in the PCR. Elevated levels of IL-6 cDNA and, to a lesser extent, interferon gamma were detected in three of three nonhuman primates with persistent infection with B burgdorferi, whereas the brains of three uninfected animals and undetectable levels of gene expression of these cytokines. These data support the hypothesis that cytokines such as IL-6 are important amplification molecules for CNS inflammation in Lyme neuroborreliosis. PMID- 9222184 TI - Cognitive, behavioral, and motor effects of the NMDA antagonist ketamine in Huntington's disease. AB - BACKGROUND: Excitotoxicity may contribute to neuronal degeneration in Huntington's disease (HD). N-methyl-D-aspartate (NMDA) receptor antagonists can prevent neuronal degeneration caused by excitotoxicity, but their effects in HD patients are not known. METHODS: We investigated the acute cognitive, behavioral, and motor effects of the NMDA-receptor antagonist ketamine in HD patients. Double blind infusions of 0.10, 0.40, and 0.60 mg/kg/hr ketamine were given to 10 HD patients on one test day and compared with placebo infusions on a second, identical testing day. Linear mixed-effects models and randomization tests were used to identify whether, and at which dose, a significant change from baseline occurred in outcome variables. RESULTS: We demonstrated that ketamine is well tolerated at low and intermediate subanesthetic doses. Intermediate ketamine doses produced specific decline in memory and verbal fluency. Higher subanesthetic doses caused a significant increase in psychiatric symptoms and impairment of eye movements. CONCLUSIONS: These results describe the spectrum of clinical effects produced by increasing NMDA receptor blockade in HD patients. The clinical effects appearing with higher levels of NMDA receptor blockade can identify the range of doses used in clinical trials of NMDA receptor antagonists. PMID- 9222185 TI - Clinical evaluation of pramipexole in advanced Parkinson's disease: results of a double-blind, placebo-controlled, parallel-group study. AB - We compared the efficacy, safety, an tolerability of pramipexole, an aminobenzathiazol-derived dopamine agonist with novel properties, with those of placebo in advanced PD patients with motor fluctuations under levodopa treatment. Pramipexole improved motor function of patients during "on" and "off" periods, decreased the time spent in "off" periods, reduced the severity of "off" periods, decreased disability and PD severity during "on" and "off" periods, as assessed by the Unified Parkinson Disease Rating Scale, and permitted a reduction in levodopa dosage. Adverse effects related to the central nervous system were similar to those reported with other dopamine agonists, and the gastrointestinal and cardiovascular tolerability of the compound was satisfactory. PMID- 9222186 TI - Tactile spatial acuity and roughness discrimination: impairments due to aging and Parkinson's disease. AB - We used gratings of alternating ridges and grooves in a quantitative psychophysical investigation of tactile perception in patients with Parkinson's disease (PD) and age-matched normal controls. The groove width required for threshold discrimination of grating orientation was 25% higher in the control subjects compared to younger individuals studied previously (p = 0.004), indicating a small but significant decline in tactile spatial acuity with age. Relative to age-matched controls, patients with PD showed a twofold increase in the tactile spatial threshold (p = 3.07 x 10(-8), with somewhat greater impairment on the side more affected clinically (p = 0.03). Testing with the forearm prone, as compared to supine, produced a small improvement in the acuity of patients (p = 0.01) but not controls (p = 0.26). PD patients were also impaired in tactually discriminating grating roughness: their difference limens were over three times higher than those of controls (p = 5.74 x 10(-5)) for gratings differing in groove width, and over twice as high (p = 0.0003) for gratings differing in ridge width. We conclude that PD significantly impairs performance on these tactile tasks. PMID- 9222187 TI - Paroxysmal dystonic choreoathetosis linked to chromosome 2q: clinical analysis and proposed pathophysiology. AB - We describe clinical features of a large Polish-American kindred in which autosomal-dominant, paroxysmal dystonic choreoathetosis (PDC) was linked to a locus on chromosome 2q. Episodes of generalized dystonia and choreoathetosis involving the face and all extremities began in early childhood, lasted for 30 minutes to several hours, and occurred up to several times each week. There was no interruption of consciousness and EEGs were normal during the episodes. Paroxysmal dyskinesia occurred at rest both spontaneously and following caffeine or alcohol consumption. Neurologic examinations were normal between attacks. The cause of PDC is unknown. We deduced a model of PDC pathophysiology from analyzing neurophysiologic effects of alcohol and caffeine (which provoke attacks of PDC), the variably beneficial effects of levodopa-carbidopa, and the occurrence of dystonia and paroxysmal dyskinesia in biopterin synthesis disorders. We propose that nigrostriatal neurons in PDC patients have either marginally deficient dopamine synthesis or excessive alcohol- and caffeine-induced dopamine release; and that following alcohol- and caffeine-induced dopamine release, PDC patients experience a period of dopamine deficiency. PMID- 9222188 TI - Dopaminergic function in patients with posttraumatic parkinsonism: an 18F-dopa PET study. AB - Posttraumatic encephalopathy (PTE) is characterized by a combination of upper motor neuron, basal ganglia, cerebellar, and psychiatric disturbances. A "striatal" variant, with predominant posttraumatic parkinsonism (PTP), is uncommon and may be difficult to distinguish from idiopathic Parkinson's disease (PD). We report the clinical characteristics of six patients clinically presumed to have PTP who were investigated with 18F-dopa PET. We compared their findings with those of age-matched controls and a group of idiopathic PD patients without a history of head trauma. The PTP patients showed a uniform 40% reduction (p < 0.0001) of mean 18F-dopa uptake in caudate and putamen compared with controls. Their mean putamen uptake was significantly higher than that seen in the PD group (p < 0.004) while mean caudate uptake was lower. The PTP mean caudate:putamen Ki ratio (0.97) was similar to that of controls (1.10) but significantly lower than that of the PD group (1.90) (p < 0.0001). These results suggest that although PTP may appear clinically similar to PD, 18F-dopa PET may help to differentiate it in vivo by demonstrating uniform nigrostriatal involvement as opposed to relative sparing of caudate function. These data also provide support for the view that delayed neurologic sequelae may follow cumulative head trauma. PMID- 9222189 TI - Human response to botulinum toxin injection: type B compared with type A. AB - Despite the clinical potential of botulinum toxin type B (BTXB) for treating focal dystonia, hemifacial spasm, and other movement disorders, particularly in those resistant to botulinum toxin type A (BTXA), no objective human data exist to compare the muscle paralysis resulting from these two botulinum toxin subtypes. To objectively compare the human muscle paralysis resulting from intramuscular injections of BTXB with that from BTXA, we measured the extensor digitorum brevis (EDB) M wave amplitude four times before and six times after injection with 17 different doses of BTXB (from 1.25 to 480 units) in 17 healthy volunteers. This established a dose-response curve that we compared with the previously published BTXA dose-response curve. After the establishment of the dose-response curve, we injected 10 new volunteers with five different doses of BTXB and BTXA measuring EDB M wave amplitude 4 times before and 13 times over 57 weeks after injection. The volunteers were randomized by dose and received BTXA and BTXB in opposite EDB muscles. The effect of the toxin in all volunteers was expressed as percent decline in M wave amplitude postinjection (% paralysis). The maximal paralysis 2 weeks postinjection with 320 to 480 mouse units (MU) of BTXB was 50 to 75%, whereas maximal paralysis was 70 to 80% with 7.5 to 10 MU of BTXA. Postexercise M wave facilitation on day 9 postinjection averaged 63% for BTXB and 20% for BTXA. Seven weeks postinjection, BTXB-induced paralysis had improved by 66% with complete improvement by 11 weeks postinjection, whereas BTXA-induced paralysis had improved by only 6% at 7 weeks, and at 57 weeks postinjection 22% of the original muscle paralysis was still present. Thus, human muscle paralysis resulting from BTXB injection is not as complete or long-lasting as that resulting from BTXA. PMID- 9222190 TI - Prevalence of headaches in a Chinese elderly population in Kinmen: age and gender effect and cross-cultural comparisons. AB - OBJECTIVE: To investigate the prevalence of headaches in a Chinese elderly population. BACKGROUND: There are few headache surveys in the elderly. Previous studies have shown a low headache prevalence in Chinese. METHODS: TARGET POPULATION: eligible registered residents > or = 65 years old (N = 2,003) in two townships of Kinmen Island on August 1, 1993. All participants completed a headache questionnaire and underwent clinical evaluation and examination by a neurologist. Headache diagnoses were made according to the International Headache Society, 1988. RESULTS: 1,533 persons (77%) participated in the study, of whom 584 (38%) had at least one episode of headache in the previous year. One-year prevalence of migraine was 3.0%, and tension-type headache, 35%. The prevalence of migraine, but not tension-type headaches, continued to decline with age in the elderly. Life-time prevalence of "incapacitating headache" was 10%, and that of migraine, 5.2%. Forty-two percent of migraineurs stopped having migraine before this survey. In comparison with "10 years ago" 8% participants felt their current headaches were worse, 25% better, and 67%, no change, with a net improvement of 17%. CONCLUSIONS: We have reported the highest headache prevalence among different Chinese elderly populations, but these were still lower than those reported from Western series. More than half of the elderly life-time migraineurs still had attacks of migraine. Severe headaches, including migraine but not tension-type headaches, declined with age. PMID- 9222191 TI - Clinical susceptibility to migraine with aura is modified by dopamine D2 receptor (DRD2) NcoI alleles. AB - Migraine has a major genetic component. Although most recent scientific studies have focused on the role of 5-hydroxytryptamine and neuropeptides in migraine, dopaminergic systems are also implicated in the pathogenesis. Therefore, the dopamine D2 receptor (DRD2) was analyzed as a candidate gene since antagonists of this receptor have been reported to be effective in the acute treatment of migraine. Individuals with migraine with aura (n = 52) have an increased frequency (0.84) of the DRD2 NcoI C allele (chi-square = 6.47; p < 0.005) compared with control individuals (n = 121; C allele frequency = 0.71). Individuals with migraine without aura (n = 77) showed the same DRD2 T allele frequency (0.70) as the control group. Migraine with aura was present in 27% of the C/C individuals, 16% of the C/T individuals, and 5.2% of the T/T individuals. These data suggest that activation of the DRD2 receptor plays a modifying role in the pathophysiology of migraine with aura. As a result, these data provide a molecular rationale for the documented efficacy of DRD2 antagonists in the treatment of migraine with aura. PMID- 9222192 TI - Peripheral neuropathy in children with HIV infection. AB - Peripheral neuropathy is infrequently reported in children with HIV infection, but may be underrecognized. To provide a better understanding of the patterns of peripheral neuropathy in these children, we surveyed the charts of 50 children with HIV infection referred to the EMG laboratory at the National Institutes of Health for evaluation of suspected peripheral neuropathy. Twelve children had an abnormal nerve conduction study. The findings suggested a distal sensory or sensorimotor axonal neuropathy in seven children, median nerve compression at the carpal tunnel in three, a demyelinating neuropathy in one child, and a lumbosacral polyradiculopathy in one adolescent. Distal symmetric polyneuropathy occurred mostly in older-aged children. PMID- 9222193 TI - Intrathecal administration of recombinant human superoxide dismutase 1 in amyotrophic lateral sclerosis: a preliminary safety and pharmacokinetic study. AB - We undertook a safety and pharmacokinetic study of intrathecal (i.t.) recombinant human superoxide dismutase (rhSOD1). We administered rhSOD1 as an acute bolus in three sheep and 16 human subjects with amyotrophic lateral sclerosis (ALS). Two sheep received chronic i.t. infusion of rhSOD1 (one at 17.7 mg per day, the second at 38.0 mg per day) for six months. Two of the 16 subjects had familial ALS and mutations in the gene for Cu/Zn SOD1. They both received i.t. infusion of rhSOD1 (5 to 10 mg per day) for 3 to 6 months. Intrathecal rhSOD1 administration was safe. Bolus i.t. administration of 0.25 mg rhSOD1 in sheep revealed a mean elimination half-life of 0.4 (SD +/- 0.06) hours, clearance of 12.2 +/- 3.2 ml per hour, and volume of distribution of 7.3 +/- 0.9 ml. After chronic i.t. infusion, the initial alpha-phase half-life was estimated as 1.2 hours and the extended beta-phase half-life was 15.0 hours. The mean clearance rate was 25.9 ml per hour and the steady-state volume of distribution was 920.6 ml. Bolus i.t. administration of 20 micrograms of rhSOD1 in ALS subjects revealed a mean elimination half-life of 2.2 +/- 0.8 hours, clearance of 1.2 +/- 0.6 ml per hour, and volume of distribution of 3.5 +/- 0.4 ml. With chronic i.t. infusion of 5 mg per day, cerebrospinal SOD1 levels increased approximately fortyfold. We detected no benefit of this treatment in the two patients with familial ALS. PMID- 9222194 TI - Maladaptive neural compensatory mechanisms in Bell's palsy-induced blepharospasm. AB - We described four patients with Bell's palsy and blepharospasm and evaluated potential mechanisms that may be responsible for an apparent association between the two disorders. Eyelid movements in spontaneous blinks were studied by the search coil technique in patients with this novel disorder. Kinematic analyses documented bilateral eyelid spasm subsequent to unilateral Bell's palsy. The temporal interval between the onset of palsy and onset of blepharospasm was highly variable (weeks to > 20 years). Changes in the relationship between spontaneous blink peak velocity and amplitude, the main sequence, shared features previously found in uncomplicated Bell's palsy and blepharospasm patients. Furthermore, as in patients with typical Bell's palsy and idiopathic blepharospasm, both normal blinks and spasms were conjugate in spite of interocular differences in blink amplitude/peak velocity. We suggest that there is a correlation between the eyelid palsy and subsequent blepharospasm, and have designated this potentially new disease entity as Bell's palsy-induced blepharospasm. We propose a two-stage model for Bell's palsy-induced blepharospasm in which blink adaptive systems may produce the maladaptive consequence of eyelid spasms. PMID- 9222195 TI - Longitudinal assessment of diabetic polyneuropathy using a composite score in the Rochester Diabetic Neuropathy Study cohort. AB - Because there are little satisfactory data on change in severity of diabetic polyneuropathy (DP) over time from study of population-based cohorts of diabetic patients in epidemiologic surveys of DP, it is difficult to predict outcome or morbidity or to identify risk factors; it is also difficult to estimate statistical power for use in controlled clinical trials. In this longitudinal study of almost 200 patients from the Rochester Diabetic Neuropathy Study (RDNS) cohort, we assess which symptoms, clinical examinations, tests, or combinations of examinations and tests (composite scores) are best used as minimal criteria for the diagnosis of DP and as a quantitative measure of severity of DP. An abnormality (> or = 97.5th percentile) of a composite score that included the Neuropathy Impairment Score of the lower limbs plus seven tests (NIS(LL)+7 tests), was a better minimal criteria for DP than clinical judgment alone or previously published minimal criteria. First, it provided a more comprehensive assessment of neuropathic impairment. Second, it avoided the overestimated frequency of DP when the minimal criteria for DP was any one or two abnormalities from multiple measurements. Minimal criteria using nerve conduction and reduced heart beat response to deep breathing identified approximately twice as many patients with DP than did clinical examination and vibration detection threshold using CASE IV. This difference could be used to subclassify state 1 DP. Although various individual measures of DP, for example, vibration detection threshold (as evaluated by CASE IV and the 4, 2, and 1 stepping algorithm [see text]), were good measures of worsening, the composite score NIS(LL)+7 tests (assessing neuropathic impairment) was much better at showing monotone worsening. Using this composite score, the average diabetic patient in the RDNS worsened by 0.34 points per year, whereas patients with diabetic polyneuropathy worsened by 0.85 points per year. On the assumption that a therapeutic agent may prevent worsening of DP but not cause improvement, controlled clinical trials of patients with DP would need to be conducted for a period of 3 years to achieve a meaningful change of 2 NIS points (the level of abnormality considered by a Peripheral Nerve Society consensus group to be clinically meaningful). PMID- 9222196 TI - Sensory ataxic neuropathy as the presenting feature of a novel mitochondrial disease. AB - Four unrelated patients presented with a severe sensory ataxic neuropathy in association with dysarthria and chronic progressive external ophthalmoplegia. Electrophysiologic and pathologic studies showed severe axonal loss disproportionately affecting sensory nerves. Molecular genetic analysis revealed multiple mitochondrial DNA deletions in muscle and peripheral nerve. Sensory ataxic neuropathy may be the predominant and presenting manifestation of a mitochondrial disorder, and a mitochondrial etiology should be included in its differential diagnosis. The triad of sensory ataxic neuropathy, dysarthria, and ophthalmoparesis (SANDO) may represent a novel mitochondrial disease associated with multiple mitochondrial DNA deletions. PMID- 9222197 TI - Reversible MRI findings in a patient with Hashimoto's encephalopathy. AB - Diffuse subcortical MRI signal abnormalities were seen during a subacute exacerbation in a patient with Hashimoto's encephalopathy. The patient had an excellent clinical response to corticosteroids. Clinical recovery paralleled normalization of MRI abnormalities and lowering of thyroid microsomal antibody titer. PMID- 9222198 TI - The diagnosis of 'essential palatal tremor'. AB - We report a patient with a 30-year history of progressive, involuntary movements of the left ear and clicking sounds in both ears for 2 years. The patient had rhythmic contractions of the tensor veli palatini muscle and could relieve palatal movements and ear clicks, but not ear movements, by pressing a pillow against the left ear or by finger pressure on the tragus or the retroauricular region. We discuss the significance of these sensory tricks and the nosology of the long-standing ear movements within a classification of essential and symptomatic palatal tremor. Current diagnostic criteria for both types of palatal tremor may not cover some atypical cases such as ours. PMID- 9222200 TI - Assessment of hand function in a patient with chronic sensory demyelinating neuropathy. AB - A 60-year-old man presented with progressive large fiber sensory loss in the right first three fingers and, to a lesser extent, in both fourth and fifth fingers. Electrophysiologic studies were characteristic of chronic sensory demyelinating polyneuropathy, a variant of chronic inflammatory demyelinating polyneuropathy. Plasma exchange was unsuccessful, but intravenous immunoglobulin (IVIG) led to complete recovery of sensation for 2 months, although neurophysiologic abnormalities persisted. A battery of noninvasive tests to measure hand grip strength, tactile sensation at the fingertips, and motor control of prehension during precision grip revealed marked abnormalities in the right hand before IVIG. One month after IVIG, all test results had normalized, but they returned to pretreatment levels after 3 months. Functional evaluation of the hand may be a sensitive method to objectively quantify loss of and changes in cutaneous mechanoreceptor function of the fingers in large fiber sensory neuropathy. PMID- 9222199 TI - High-frequency stimulation of the globus pallidus for the treatment of Parkinson's disease. AB - Long-term treatment of Parkinson's disease (PD) with levodopa is complicated by the development of motor fluctuations and dyskinesias. Posteroventral pallidotomy can improve tremor, bradykinesia, rigidity, and dyskinesias in PD. We performed chronic stimulation of the globus pallidus (CSGP) to duplicate the positive results of pallidotomy with reduced risk of permanent neurologic deficit in patients with advanced PD. The lead for CSGP was stereotactically implanted with the aid of microelectrode recordings in the globus pallidus pars interna. An electrical pulse generator was implanted in the subclavicular region. Stimulation settings were adjusted by computer. Five PD patients (four men, one woman) with disabling symptoms were enrolled. Three of the patients had bilateral implants. At 3 months following the last implant, four patients rated themselves as markedly improved, and one patient was moderately improved. The amount of time in the "on" state increased from 21% at baseline to 65% at 3-month follow-up (p < 0.05). There was a significant improvement in all subscales of the UPDRS (p < 0.05). One patient had an asymptomatic intracranial bleed, one patient had transient hemiparesis during surgery with stimulation, and one patient required surgical repositioning of the lead. Adverse effects caused by stimulation were minimal. CSGP is a safe and effective procedure in PD patients with motor fluctuations and dyskinesias. PMID- 9222201 TI - A case of Hodgkin's lymphoma producing neuromyotonia. AB - The syndrome of neuromyotonia produces muscle stiffness, cramps, and frequently, excessive sweating. Most cases are idiopathic, but some are associated with neoplasms, especially immune cell cancers. Voltage-gated potassium channels may be the target of an autoantibody attack in idiopathic generalized neuromyotonia (Isaacs' syndrome). The cases associated with neoplasms may also have an autoimmune etiology. We report the first case of neuromyotonia as the presenting feature of Hodgkin's lymphoma and propose a paraneoplastic mechanism that would link the purported autoimmune etiology in Isaacs' syndrome with the cancer associated cases. PMID- 9222202 TI - Xp21 muscular dystrophy due to X chromosome inversion. AB - Two brothers with Duchenne muscular dystrophy have an inversion of the X chromosome, 46, Y, inv(X) (p11.2p21.2). Because their mother is an unaffected carrier of the inversion, this confirms that maternal passage of a structurally abnormal X chromosome can cause dystrophinopathy in males. Our experience suggests that as well as molecular genetic analysis, karyotyping can be useful in Xp21 muscular dystrophy. PMID- 9222203 TI - Prenatal brachial plexus paralysis. AB - An 18-day-old child with a history of difficult breech delivery presented with wasting and weakness of C5-6-innervated muscles. The EMG examination performed the same day showed high-voltage polyphasic motor unit potentials without abnormal spontaneous activity. Both the EMG pattern and the clinical features suggest an injury taking place several weeks before delivery. This presumption seems confirmed when compared with findings in our series of 100 EMG examinations in 78 children with diagnosis of Erb's palsy. Because of both clinical and medicolegal implications, early EMG testing is advisable in all patients with congenital brachial paralysis. PMID- 9222204 TI - Progressive multifocal leukoencephalopathy presenting as human immunodeficiency virus type 1 (HIV)-associated dementia. AB - Progressive multifocal leukoencephalopathy (PML) is a demyelinating disorder of the CNS that usually causes hemiparesis or hemianopsia. Dementia occurs in combination with other neurologic abnormalities. We report a human immunodeficiency virus type 1 (HIV)-infected man whose only manifestation of proven PML was dementia that was clinically indistinguishable from HIV-associated dementia. PMID- 9222205 TI - Resolution of a brainstem abscess through antituberculous therapy. AB - We describe an immunocompetent patient with a solitary brainstem abscess that responded to antituberculous therapy. Although prompt surgical therapy has been advocated, the possibility of medical resolution of brainstem tuberculous abscesses should be considered. PMID- 9222206 TI - Loss of merlin expression in sporadic meningiomas, ependymomas and schwannomas. AB - Neurofibromatosis 2 (NF2) is an inherited disorder in which affected individuals develop schwannomas and meningiomas. NF2 is mapped to chromosome 22q in a region where frequent loss of heterozygosity also occurs in sporadic meningiomas, ependymomas, and schwannomas. Using NF2 protein (merlin or schwannomin)-specific antibodies, 11 of 14 sporadic schwannomas, three of eight sporadic ependymomas, and 16 of 19 sporadic meningiomas demonstrated significantly reduced or absent merlin expression, suggesting that NF2 may be involved in the pathogenesis of these sporadic tumors. PMID- 9222207 TI - Heterogeneous clinical presentation of the mtDNA NARP/T8993G mutation. AB - To obtain a better molecular definition of patients with syndromic retinitis pigmentosa, we screened for mitochondrial DNA (mtDNA) alterations of the two ATPase genes and 22 tRNA-coding sequences in 10 patients whose features resembled NARP (neuropathy, ataxia, and retinitis pigmentosa) syndrome. In two patients, one of whom showed features mimicking Kearns-Sayre syndrome, we identified a heteroplasmic T8993G mutation (average 80%) in the mitochondrial ATPase 6 gene. There was no mutated mtDNA in muscle and leukocytes from the mother of one patient or in leukocytes from his brother, suggesting a rapid segregation of the mutated nucleotide. MtDNA analysis should be considered in the differential diagnosis of patients with syndromic retinitis pigmentosa. PMID- 9222208 TI - The effect of imprecise repositioning on lesion volume measurements in patients with multiple sclerosis. AB - In this study, we evaluated the effect of imprecision in patient repositioning encountered in real life on multiple sclerosis (MS) lesion volumes measured from MRIs. We also evaluated two putative methods for reducing the variability in these lesion volume measurements: first, a reduction of slice thickness (from the conventional 5 mm to 3 mm) and second, the application of a new repositioning technique based on the use of head immobilization shells. We evaluated the errors in lesion volume by scanning 10 patients a total of four times using the two slice thicknesses and two repositioning methods (conventional and using a head immobilization shell). The mean absolute percentage difference between two corresponding scans was 6.8% (range, 1.24 to 11%) using conventional slice thickness and repositioning, 4.1% (range, 0.7 to 5.56%) using conventional slice thickness and head immobilization shells, 2.6% (range, 0.8 to 6.66%) using the conventional repositioning technique and 3-mm slice thickness, and 1.4% (range, 0.2 to 6.14%) using slice thickness of 3 mm and head immobilization shells. These mean absolute differences were significantly different (p = 0.0008). Our results indicate that the effect of repositioning errors of the order of those that can be encountered in the daily life situation of clinical trials affects significantly lesion load measurements in MS and that the combined use of thinner slices and more accurate repositioning techniques can markedly improve the reproducibility of such measurements. PMID- 9222209 TI - Assessment of digital EEG, quantitative EEG, and EEG brain mapping: report of the American Academy of Neurology and the American Clinical Neurophysiology Society. AB - A. Digital EEG is an established substitute for recording, reviewing, and storing a paper EEG record. It is a clear technical advance over previous paper methods. It is highly recommended. (Class III evidence, Type C recommendation). B. EEG brain mapping and other advanced QEEG techniques should be used only by physicians highly skilled in clinical EEG, and only as an adjunct to and in conjunction with traditional EEG interpretation. These tests may be clinically useful only for patients who have been well selected on the basis of their clinical presentation. C. Certain quantitative EEG techniques are considered established as an addition to digital EEG in: C.1. Epilepsy: For screening for possible epileptic spikes or seizures in long-term EEG monitoring or ambulatory recording to facilitate subsequent expert visual EEG interpretation. (Class I and II evidence, Type A recommendation as a practice guideline). C.2. OR and ICU monitoring: For continuous EEG monitoring by frequency-trending to detect early, acute intracranial complications in the OR or ICU, and for screening for possible epileptic seizures in high-risk ICU patients. (Class II evidence, Type B recommendation as a practice option). D. Certain quantitative EEG techniques are considered possibly useful practice options as an addition to digital EEG in: D.1. Epilepsy: For topographic voltage and dipole analysis in presurgical evaluations. (Class II evidence, Type B recommendation). D.2. Cerebrovascular Disease: Based on Class II and III evidence, QEEG in expert hands may possibly be useful in evaluating certain patients with symptoms of cerebrovascular disease whose neuroimaging and routine EEG studies are not conclusive. (Type B recommendation). D.3. Dementia: Routine EEG has long been an established test used in evaluations of dementia and encephalopathy when the diagnosis remains unresolved after initial clinical evaluation. In occasional clinical evaluations, QEEG frequency analysis may be a useful adjunct to interpretation of the routine EEG when used in expert hands. (Class II and III evidence as a possibly useful test, Type B recommendation). E. On the basis of current clinical literature, opinions of most experts, and proposed rationales for their use, QEEG remains investigational for clinical use in postconcussion syndrome, mild or moderate head injury, learning disability, attention disorders, schizophrenia, depression, alcoholism, and drug abuse. (Class II and III evidence, Type D recommendation). F. On the basis of clinical and scientific evidence, opinions of most experts, and the technical and methodologic shortcomings, QEEG is not recommended for use in civil or criminal judicial proceedings. (Strong Class III evidence, Type E recommendation). G. Because of the very substantial risk of erroneous interpretations, it is unacceptable for any EEG brain mapping or other QEEG techniques to be used clinically by those who are not physicians highly skilled in clinical EEG interpretation. (Strong Class III evidence, Type E recommendation). PMID- 9222210 TI - Assessment of vagus nerve stimulation for epilepsy: report of the Therapeutics and Technology Assessment Subcommittee of the American Academy of Neurology. PMID- 9222212 TI - Cerebral vasoconstriction after carotid surgery. PMID- 9222211 TI - Should spinocerebellar ataxia type 5 be called Lincoln ataxia? PMID- 9222213 TI - Amelioration of refractory dysesthetic limb pain in multiple sclerosis by gabapentin. PMID- 9222214 TI - Pure alexia could not be a disconnection syndrome. PMID- 9222215 TI - Anemia associated with lamotrigine. PMID- 9222216 TI - Inflammatory trigeminal sensory neuropathy. PMID- 9222217 TI - Cerebral infarction associated with Kearns-Sayre syndrome. PMID- 9222218 TI - Degos' disease. PMID- 9222219 TI - Electrophysiology in locked-in syndrome. PMID- 9222220 TI - Prion in progressive subcortical gliosis revisited. PMID- 9222221 TI - Diet and Parkinson's disease. PMID- 9222222 TI - Mechanical ventilation. PMID- 9222223 TI - Mechanical ventilation. PMID- 9222224 TI - EMFs and Alzheimer's disease. PMID- 9222225 TI - Assessment of health-related quality of life after corneal transplantation. AB - PURPOSE: To determine the extent to which commonly used clinical measures of corneal transplantation outcome are related to aspects of visual function and health-related quality of life. METHODS: In a cross-sectional validation study, ophthalmic examination information was collected by chart review of, and health related quality of life information was collected by telephone contact with, patients (n = 77) undergoing routine follow-up examinations at least 1 year after corneal transplantation. A questionnaire that included the VF-14 and SF-36 instruments was completed for each participant. Associations between clinical and questionnaire outcomes were evaluated using analysis of variance and regression analyses. RESULTS: When the best-corrected visual acuity of both eyes was evaluated, there was a positive association (P < .0001) of visual acuity with the VF-14 score and with the following SF-36 scales: role limitations because of emotional problems (P = .04), emotional well-being (P = .08), and social functioning (P = .02). Those with a high degree of keratometric astigmatism showed an impact on social functioning (P = .005). Upon regression analysis, the single most important factor associated with the patients' reported visual function was their visual acuity in the better eye, followed by the extent of keratometric astigmatism. CONCLUSIONS: These findings demonstrate a high degree of criterion validity in using the VF-14 instrument to assess the outcome of corneal transplantation. Application of the more generic SF-36 measure shows effects of visual disability on other aspects of corneal transplant patients' health status, including their emotional and social functioning. PMID- 9222226 TI - Macular corneal dystrophy in Saudi Arabia: a study of 56 cases and recognition of a new immunophenotype. AB - PURPOSE: To determine the immunophenotype or immunophenotypes of macular corneal dystrophy in Saudi Arabia. METHODS: We studied 56 cases of macular corneal dystrophy. Tissue from 60 corneal transplant buttons was stained by the avidin biotin complex method using an anti-keratan sulfate monoclonal antibody. The serum antigenic keratan sulfate was measured in 23 of the 56 patients, four unaffected relatives, and 13 individuals with chronic actinic keratopathy. Serum and corneal tissue were studied in 17 of the 50 affected individuals with corneal transplant material. RESULTS: Thirty-five corneas (58.3%) of 29 of 50 patients did not react with anti-keratan sulfate monoclonal antibody. The stroma and abnormal intracellular and extracellular corneal accumulations reacted with anti keratan sulfate monoclonal antibody in seven corneas (11.7%). The stroma in the other 18 corneas (30.0%) from 15 patients did not react with the anti-keratan sulfate monoclonal antibody, but corneal fibroblasts did. Twenty-one of the 23 patients with macular corneal dystrophy had no detectable serum antigenic keratan sulfate (< 9 ng/ml); two had values of 12 and 51 ng/ml, respectively, and their corneal stroma and abnormal accumulations reacted with anti-keratan sulfate monoclonal antibody. CONCLUSIONS: We detected macular corneal dystrophy type IA, a new immunophenotype characterized by the lack of detectable antigenic keratan sulfate in the serum (< 9 ng/ml), and a corneal stroma that did not react with the keratan sulfate monoclonal antibody but in which corneal fibroblasts did react with keratan sulfate monoclonal antibody (in 15 of 50 patients). PMID- 9222227 TI - Infectious crystalline keratopathy caused by gram-negative bacteria. AB - PURPOSE: To identify the characteristics and outcomes of infectious crystalline keratopathy caused by gram-negative bacteria. METHODS: We reviewed all patients treated at a university eye center for infectious crystalline keratopathy from 1978 through 1995 and performed a nested case-comparison study by comparing patients with keratitis caused by gram-negative rods and those with keratitis caused by gram-positive cocci. RESULTS: Eighteen patients (mean age +/- SD, 59 +/ 17 years) displayed unilateral culture-positive infectious crystalline keratopathy. Among 18 eyes with crystalline keratopathy, five occurrences (28%) were caused by gram-negative rods (Acinetobacter lwoffi, Citrobacter koseri, Enterobacter aerogenes, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia), 10 (55%) were caused primarily by gram-positive cocci, and three (17%) were caused primarily by yeasts. Four cases grew two different isolates. No significant difference in predisposing factors, clinical appearance, or visual outcome was found between infections caused by gram-negative bacteria and those caused by gram-positive bacteria. CONCLUSIONS: Gram-negative bacteria can cause infectious crystalline keratopathy but have no distinguishing features from infectious crystalline keratopathy caused by streptococci and other gram-positive bacteria. Appropriate laboratory evaluation is therefore necessary to guide specific antimicrobial therapy. PMID- 9222228 TI - Blinking is controlled primarily by ocular surface conditions. AB - PURPOSE: To investigate the relation between blinking and ocular surface conditions and to introduce and examine a new index, the maximum blink interval. METHODS: In a prospective study, the blink rate of subjects under relaxed conditions was determined from a video recording taken by a hidden observer. The maximum blink interval was defined as the longest time subjects can avoid blinking without feeling uncomfortable. RESULTS: Significant changes in the blink rate and maximum blink interval were induced by factors that directly or indirectly affect the ocular surface: topical anesthesia, changing exposed ocular surface area, and wind. Moreover, the blink rate and maximum blink interval were significantly different in dry eye patients compared with healthy volunteers, with the values of the former approaching the values of the latter after use of artificial tears. The maximum blink interval was decreased by the same factors that increased the blink rate, and there was a significant inverse correlation between blink rate and maximum blink interval. Use of video display terminals was associated with decreased maximum blink interval and, hence, the development of dry eye symptoms. CONCLUSIONS: There was an important association among blink rate, maximum blink interval, and ocular surface conditions. The blink rate and our newly introduced measurement, the maximum blink interval, should prove useful in assessing factors that cause dry eye. This prospective study should contribute to the understanding and treatment of dry eyes. PMID- 9222229 TI - Efficacy of topical povidone-iodine during the first week after ophthalmic surgery. AB - PURPOSE: In the first postoperative day, povidone-iodine ophthalmic solution prevents an increase in conjunctival bacterial colony-forming units and decreases the species compared with antibiotic. We sought to determine whether these beneficial effects of povidone-iodine could be sustained during the first postoperative week. METHODS: In 42 eyes of 35 consecutive patients, one or two drops of either a broad-spectrum antibiotic (polymyxin B sulfate-neomycin sulfate gramicidin) or povidone-iodine 1.25% to 2.5% were placed in the treated eye or eyes at the conclusion of surgery and three times daily during the first postoperative week. Bacterial cultures were taken from both eyes at the end of surgery before instillation of either of the eyedrops and again 1 week later. Twenty-eight untreated eyes served as a control group. RESULTS: During the first postoperative week, the number of colony-forming units and species increased in both treatment groups. Relative to the control group, both medications effectively reduced the mean number of colony-forming units at 1 week (P < .02), but their effects on colony-forming units did not significantly differ from each other (80 +/- 290 for the povidone-iodine-treated eyes and 75 +/- 90 for the antibiotic-treated eyes). At 1 week, the species count increased 281% in the antibiotic group but only 106% in the povidone-iodine group. Compared to the control group, eyes that received povidone-iodine had a significantly lower species count (P = .0097). CONCLUSION: Povidone-iodine ophthalmic solution is an alternative to postoperative topical antibiotics because of its effectiveness in controlling conjunctival bacterial colony-forming units and species, its relatively low cost, and its availability. PMID- 9222230 TI - Cataract and aqueous flare levels in patients with atopic dermatitis. AB - PURPOSE: To determine whether cataract in patients with atopic dermatitis is associated with higher levels of aqueous flare or cells. METHODS: In a prospective study, 35 consecutive patients examined during a 6-month period at the atopic dermatitis service in a university hospital underwent standardized ophthalmologic evaluations including the quantitative measurement of aqueous flare and cells by a laser flare-cell meter. RESULTS: Seven patients had bilateral cataract with anterior or posterior subcapsular opacities, or both; one patient had similar cataract in one eye and no cataract in the opposite eye; and 27 patients had no cataract in either eye. Fifteen eyes with cataract showed significantly higher levels of aqueous flare (2.1 to 33.9 photon counts per millisecond with a median of 18.0) compared with 55 eyes without cataract (2.4 to 16.0 photon counts per millisecond with a median of 9.2; Mann-Whitney U test, P = .0008). The association of cataract with higher levels of aqueous flare remained significant when only one eye (the right eye) of each patient was chosen for statistical analysis (P = .0024). CONCLUSION: Higher levels of aqueous flare caused by the breakdown of blood-aqueous barrier may contribute to the formation of cataract in patients with atopic dermatitis. PMID- 9222231 TI - Late onset of sequential multifocal bleb leaks after glaucoma filtration surgery with 5-fluorouracil and mitomycin C. AB - PURPOSE: To describe the late onset of sequential multifocal bleb leaks as a postoperative complication after filtering surgery with antimetabolites. MATERIALS: Retrospectively, 385 consecutive eyes (304 patients) undergoing trabeculectomy with 5-flurouracil (5-FU) or mitomycin C (MMC) from 1989 to 1994 were reviewed. Eyes with filtration bleb leak occurring 6 months or more after trabeculectomy were analyzed, and clinical characteristics of the filtration bleb, response to treatment, and bleb histopathology from eyes undergoing bleb excision were analyzed. RESULTS: In seven (1.8%) of 385 consecutive eyes from 304 patients undergoing glaucoma filtration surgery with 5-FU or MMC, repetitive bleb leaks in different locations of the bleb were observed from 9 to 44 months (mean, 20.4 months) after the procedure. One hundred ninety-three eyes (50%) were treated with 5-FU and the remaining eyes, with MMC. All eyes had transparent, avascular, lobular, cystic blebs. Bleb leaks occurred in five eyes treated postoperatively with subconjunctival 5-FU and in two eyes in which MMC was used intraoperatively. Three eyes (all treated with 5-FU) required surgical excision, and four eyes healed with soft contact lens, cyanoacrylate glue, or intrableb injection of autologous blood. Histopathology of the bleb leak sites demonstrated focal epithelial thinning and interruption with subjacent hypocellularity and stromal collagen degeneration. CONCLUSION: Late sequential multifocal bleb leaks may occur after glaucoma filtration surgery with administration of antimetabolites (5-FU or MMC) and are associated with epithelial break-down, hypocellularity, and stromal collagen necrosis in the filtration bleb. PMID- 9222233 TI - Anterior chamber depth in Mongolians: variation with age, sex, and method of measurement. AB - PURPOSE: To document anterior chamber depth in a Mongolian population and quantify the variation in this parameter attributable to age, sex, and method of measurement. METHOD: Depth of the anterior chamber was measured by optical pachymetry in 1,242 subjects aged 10 to 87 years. Figures for "true" anterior chamber depth were calculated by subtracting central corneal thickness from the distance between the anterior corneal epithelium and anterior lens capsule. A mode ultrasound was also used to measure the distance from anterior corneal epithelium to anterior lens capsule in 94% (942) of subjects aged 40 years and older. These ultrasound data were compared with measurements of the same distance obtained by optical pachymetry. RESULTS: Mean anterior chamber depth in women was more shallow than in men of all ages (ANOVA, P < .0001), although this difference varied according to age. Mean anterior chamber depth decreased with age and was most accurately represented by a cubic function of age. This change was maximal between the ages of 30 and 60 years and equaled 0.15 mm per decade in men and 0.21 mm per decade in women. Mean depth of the anterior chamber measured by ultrasound was significantly less than the equivalent optical measurement (difference of 0.14 mm in right eyes, P < .001). CONCLUSIONS: Mean anterior chamber depth in Mongolians decreases with age and is more shallow in women than in men. Ultrasound and optical methods of anterior chamber depth measurement yield significantly different results and are therefore not directly comparable. PMID- 9222232 TI - Asymmetries in the normal short-wavelength visual field: implications for short wavelength automated perimetry. AB - PURPOSE: To quantify short-wavelength sensitivity in normal eyes by hemifield location, eccentricity, and age. METHODS: We measured achromatic and short wavelength thresholds across visual fields covering a radius of 21 degrees of visual angle in 115 normal eyes in subjects aged 17 to 77 years and out to 30 degrees of eccentricity in an additional 57 eyes in subjects aged 22 to 80 years. RESULTS: Results indicated significantly greater sensitivity for the inferior visual field compared with the superior field (P = .001). The amount of asymmetry increased with eccentricity (P = .001) but not with age (P = .357). A temporonasal field asymmetry was noted at the most eccentric points of the 30 degree field (P = .001) but not at 21 degrees (P = .821). CONCLUSIONS: In addition to increasing our understanding of normal retinal function, these results have implications for basic research in comparison with results of studies using different retinal locations to assess short-wavelength sensitivity and for clinical practice, where short-wavelength sensitivity is used to diagnose and manage a number of diseases, including glaucoma, diabetic retinopathy, and acquired immunodeficiency syndrome (AIDS)-related vision loss. PMID- 9222234 TI - Rhegmatogenous retinal detachment in patients with cytomegalovirus retinitis: the Foscarnet-Ganciclovir Cytomegalovirus Retinitis Trial. The Studies of Ocular Complications of AIDS (SOCA) Research Group in Collaboration with the AIDS Clinical Trials Group (ACTG). AB - PURPOSE: To determine the incidence and risk factors for rhegmatogenous retinal detachment in a population of patients with newly diagnosed cytomegalovirus retinitis. METHODS: Analysis of selected baseline and time-dependent data on patients enrolled in a multicenter, prospective, randomized, controlled clinical trial of therapy with foscarnet vs ganciclovir. RESULTS: In 316 eyes with cytomegalovirus retinitis at baseline, the risk of rhegmatogenous retinal detachment in an eye involved by cytomegalovirus retinitis was 18.9% at 6 months (95% confidence interval [CI], 14.0% to 23.8%) and 37.9% at 1 year (95% CI, 30.5% to 45.3%). Retinal detachment was not associated with the type of anticytomegalovirus therapy (intravenous foscarnet or ganciclovir) to which the patient was assigned. Extent of retinal involvement by cytomegalovirus retinitis, higher patient age, and lower CD4+ T-cell counts were associated with an increased risk of retinal detachment; myopia was not. CONCLUSIONS: Retinal detachment in patients with cytomegalovirus retinitis is unrelated to the type of intravenous therapy used or to refractive error. The median time to retinal detachment in an involved eye with cytomegalovirus retinitis and free of retinal detachment at baseline was 18.2 months. Strategies to reduce the extent of retinitis and possibly the number of reactivations may reduce the incidence of retinal detachment. PMID- 9222235 TI - Uveitis associated with an epidemic outbreak of leptospirosis. AB - PURPOSE: To define uveitis associated with leptospirosis in a clinical setting. METHODS: We present the clinical features of 73 consecutive cases of uveitis linked clinically to an outbreak of systemic leptospirosis in patients with antibodies to Leptospira species who were examined from January to September 1994. RESULTS: In 73 patients, the pattern of ocular involvement was unilateral in 35 and bilateral in 38. Panuveitis was seen in 106 eyes (95.5%), retinal periphlebitis in 57 eyes (51.4%), and hypopyon in 14 eyes (12.6%). Anterior uveitis alone without hypopyon was observed in three eyes (2.7%), whereas vitreous inflammatory reaction alone was seen in two eyes (1.8%). Sixty of 73 patients (82.2%) or 91 of 111 eyes (82.0%) were followed up for 8 months. Final visual acuity was 6/6 (20/20) in 47 eyes (52%) and improved during treatment, although not up to 6/6, in 15 eyes (16%). Twenty-eight eyes (31%) maintained same vision, and one eye showed deterioration of vision. CONCLUSION: Uveitis associated with leptospirosis may manifest as unilateral or bilateral uveitis, anterior uveitis, or panuveitis. The prognosis is generally good in this entity, even when the inflammation is severe. Awareness of this disease in endemic areas is important in order to differentiate it from other uveitic entities, especially in young male patients in whom other immunologic uveitides are also common. PMID- 9222236 TI - Lacrimal punctal occlusion for the treatment of superior limbic keratoconjunctivitis. AB - PURPOSE: To test the hypothesis that superior limbic keratoconjunctivitis is caused by insufficient tear supply to the superior keratoconjunctiva. METHODS: We used cautery and sutures to permanently occlude the lacrimal puncta of 11 patients (22 eyes) with superior limbic keratoconjunctivitis for whom topical treatment was ineffective. RESULTS: All 11 patients (22 eyes) responded favorably to lacrimal punctal occlusion. After lacrimal punctal occlusion, rose bengal and fluorescein staining (both on a scale of 0 [no staining] to 9 [complete staining]) were reduced (mean +/- SD, 2.7 +/- 1.6 to 1.1 +/- 1.8 and 1.4 +/- 1.2 to 0.4 +/- 0.8, respectively). Impression cytology disclosed improvement of squamous metaplasia in the superior conjunctiva as well as increased goblet cells in nine of 13 eyes (69%) examined. Subjective symptoms improved in all 22 eyes (100%). CONCLUSIONS: Improvement of local tear deficiency to the superior limbic portion by punctal occlusion was an effective treatment in this small series. Superior limbic keratoconjunctivitis might be caused by the insufficient local tear supply. PMID- 9222237 TI - Visual field loss after vitrectomy for full-thickness macular holes. AB - PURPOSE: To report the results of a prospective study of the incidence of peripheral visual field loss after macular hole surgery. METHODS: Prospectively, 30 eyes of 30 consecutive patients with full-thickness macular holes operated on between December 1995 and April 1996 had preoperative and postoperative Goldmann visual field tests. The surgical procedure consisted of three-port pars plana vitrectomy, posterior hyaloid removal, nonexpansile fluid-hexafluoroethane (C2F6) exchange, and, in 19 of 30 patients, autologous platelet injection, followed by face-down positioning. RESULTS: Twenty-nine of these 30 cases were considered to be anatomic successes. Comparison of preoperative and postoperative visual fields disclosed that four patients (13%) had a peripheral scotoma, including one patient with stage 4 macular hole. Three other patients (10%) had a postoperative relative arcuate defect. Mean postoperative intraocular pressure was higher in the latter group. None of the patients complained of peripheral scotoma. CONCLUSIONS: Overall, seven of 30 patients (23%) had a postoperative visual field defect. Two categories of scotomas were observed: peripheral and relative arcuate. The cause of peripheral visual field loss is unclear. Increased intraocular pressure may be the cause of relative arcuate scotomas. PMID- 9222238 TI - Phenotypes in three Swedish families with X-linked retinitis pigmentosa caused by different mutations in the RPGR gene. AB - PURPOSE: To assess the clinical phenotypes in three Swedish families with X linked retinitis pigmentosa caused by different mutations in the RPGR gene. METHODS: Three families from different parts of Sweden, including nine patients with retinitis pigmentosa and six female carriers of X-linked retinitis pigmentosa, were examined clinically. Ophthalmologic examination included kinetic perimetry with a Goldmann perimeter using standardized objects I4e and V4e, dark adaptation final thresholds with a Goldmann-Weeker adaptometer, and full-field electroretinograms. RESULTS: The clinical findings in the patients demonstrated a severe form of retinitis pigmentosa with visual handicap early in life. Patients with a microdeletion of exons 8 through 10 of the RPGR gene had a more severe phenotype compared to the patients with single base-pair mutations in the introns 10 and 13 of the RPGR gene, resulting in splicing defects. Furthermore, heterozygous carriers in these families displayed a wide spectrum of clinical features, from minor symptoms to severe visual disability. CONCLUSION: These three families show a variable clinical phenotype resulting from different mutations in the RPGR gene. A microdeletion spanning at least parts of exons 8 through 10 seems to result in a severe phenotype compared to the splice defects. Heterozygous carriers of X-linked retinitis pigmentosa with these specific RPGR genotypes also show a variability of the phenotype; carriers with the microdeletion may be severely visually handicapped. PMID- 9222239 TI - Implications of thrombocytosis in giant cell arteritis. AB - PURPOSE: To bring attention to the implications of thrombocytosis in giant cell arteritis. METHOD: Case report. Platelet counts were measured before and after corticosteroid therapy in a patient with biopsy-proven giant cell arteritis. RESULT: Platelet counts were increased before corticosteroid therapy was begun and returned to normal levels after corticosteroid therapy was instituted. CONCLUSION: Platelet counts in a patient with giant cell arteritis may have diagnostic, therapeutic, and prognostic value. PMID- 9222240 TI - Acute lymphoblastic leukemia manifesting in an adult as a conjunctival mass. AB - PURPOSE: To describe the clinical manifestation and course of disseminated acute lymphoblastic leukemia in an adult. METHODS: Case report. Recurrence of disseminated disease was heralded by "salmon patch" involvement of the conjunctiva. The ocular tumor was treated successfully with external beam radiotherapy. RESULTS: The patient, who had a history of T-cell acute lymphoblastic leukemia, had recurrent disease involving the left eye conjunctiva. The diagnosis was confirmed by histopathologic analysis. CONCLUSIONS: Although acute lymphoblastic leukemia typically occurs in children, the tumor can masquerade as a conjunctival lymphoma in adults. PMID- 9222241 TI - Solitary extranodal anaplastic large cell lymphoma, Ki-1+, of the eyelid. AB - PURPOSE: To report a rare case of solitary anaplastic large cell lymphoma, Ki-1+, of the eyelid. METHOD: Case report. A firm ulcerated mass of the lower eyelid in a 10-year-old boy was the initial and only sign of anaplastic large cell lymphoma. RESULTS: A local excision of the mass was performed. Histologic examination disclosed large lymphoid anaplastic cells that reacted positively for T-cell markers and CD30 antigen. CONCLUSION: A solitary eyelid mass can be an initial sign of anaplastic large cell lymphoma in children. PMID- 9222242 TI - Surgical correction of incomplete cryptophthalmos in Fraser syndrome. AB - PURPOSE: To demonstrate favorable long-term visual outcome after ocular reconstruction in an infant with Fraser syndrome and with complete left cryptophthalmos. METHODS: Reconstruction of incomplete right cryptophthalmos in our patient was accomplished in a stepwise manner, beginning in the third week of life, by dissecting the eyelids from the cornea, reconstructing the conjunctival fornices with buccal mucosa, and repairing the upper lid coloboma in a flap reconstruction using the inferior eyelid margin. RESULTS: At age 3 years, the patient currently has good movement of the right eyelids when blinking, reasonable right tear function, and a visual acuity in the right eye between 20/200 and 20/360 on forced preferential looking. CONCLUSION: In selected cases of incomplete cryptophthalmos, oculoplastic and corneal surgery may result in useful vision and in good eyelid movement when blinking. PMID- 9222243 TI - Floppy eyelid syndrome in a child with chronic unilateral conjunctivitis. AB - PURPOSE: To report a 2-year-old child who had chronic unilateral conjunctivitis and spontaneous left upper eyelid eversion during sleep consistent with the floppy eyelid syndrome. METHODS: The patient's parents used a video camera to document nocturnal ectropion of the left upper eyelid. Examination demonstrated left upper eyelid swelling and left palpebral conjunctival hyperemia with papillary hypertrophy. RESULTS: All signs and symptoms of the floppy eyelid syndrome resolved with taping of the left upper and lower eyelids to close the palpebral fissure and to prevent ectropion during sleep, and with application of ocular lubricants. CONCLUSIONS: Floppy eyelid syndrome may manifest in childhood without other contributing conditions and should be considered in the differential diagnosis of chronic papillary conjunctivitis in patients at any age. PMID- 9222244 TI - Ocular leech infestation in a child. AB - PURPOSE: To describe a patient with manifestations of ocular leech infestation. METHOD: Case report. RESULTS: The ocular foreign body was identified as a leech, Limnatis nilotica, by parasitologic examination. The leech was extracted, and the patient began using topical antibiotic and cycloplegic agents. By the third day after extraction, the patient had no obvious symptoms or signs, except for a limited subconjunctival hemorrhage, and no epithelial defect on the cornea was observed. CONCLUSIONS: Ocular leech infestation should be considered in patients with a history of swimming in streams and lakes. Attention should also be given to ocular leech infestation in the differential diagnosis of ocular trauma with iris prolapse. PMID- 9222245 TI - Dominant inheritance of optic pits. AB - PURPOSE: To report the familial occurrence of optic pits and to screen the candidate PAX2 gene for mutations in this family. METHODS: Clinical family study. Standard mutation analysis of the PAX2 exons. RESULTS: Unilateral optic pits were present in three generations of one family and were inherited in an autosomal dominant fashion. No mutations in the PAX2 gene, responsible for the renal-optic coloboma syndrome, were found. CONCLUSION: Unilateral optic pits may be inherited in an autosomal dominant fashion and not in association with mutation in the PAX2 gene. PMID- 9222246 TI - Shewannela putrefaciens endophthalmitis after penetrating keratoplasty. AB - PURPOSE: To report a case of Shewannela putrefaciens endophthalmitis after penetrating keratoplasty. METHODS: Case report. Vitreous of the recipient and the preservative medium of donor cornea were cultured. RESULTS: Vitreous of the recipient eye and the donor eye corneal preservative medium both grew S putrefaciens. The patient failed to respond to intravitreal, topical, and systemic amikacin and cefotaxime. Vision was lost rapidly. Evisceration was performed. CONCLUSION: Shewannela putrefaciens should be considered as a potential pathogen contaminating donor cornea. Shewannela putrefaciens endophthalmitis is devastating and responds poorly to medical treatment. PMID- 9222247 TI - Delayed hyphema and intravitreal blood following intrableb autologous blood injection after trabeculectomy. AB - PURPOSE: To report delayed hyphema and intravitreal blood as complications following intrableb autologous blood injection after trabeculectomy. METHODS: Case report. A 44-year-old woman with hypotony and maculopathy after trabeculectomy with mitomycin C received an intrableb autologous blood injection. RESULTS: Three days after the blood injection, a hyphema formed and subsequently dispersed into the vitreous. CONCLUSIONS: Although immediate hyphema from autologous blood injection is common, hyphema may be delayed and associated with intravitreal blood. PMID- 9222248 TI - Bilateral loss of vision and macular ischemia related to Behcet disease. AB - PURPOSE: To report a 61-year-old man with sudden onset of visual loss in both eyes related to bilateral macular ischemia. METHODS: The patient underwent comprehensive ophthalmic examination including slit-lamp and fundus examination, fluorescein angiography, and visual field testing as well as biologic screening. RESULTS: Bilateral macular ischemia associated with peripheral retinal vasculitis was confirmed by angiography. We diagnosed Behcet disease by association of ocular, oral, and cutaneous involvement according to the criteria of the international study group for Behcet disease. The patient was treated with corticosteroids. CONCLUSION: In Behcet disease, sudden onset of bilateral vision loss may be associated with bilateral retinal vascular disease and macular ischemia. Prompt diagnosis and treatment with systemic corticosteroids may be beneficial. PMID- 9222249 TI - Leukocoria caused by occult penetrating trauma in a child. AB - PURPOSE: To report a child with leukocoria caused by occult penetrating trauma. METHODS: Case report. The clinical findings and surgical repair of acquired leukocoria of the right eye in a 2-year-old boy are presented. RESULTS: In the right eye, slit-lamp examination disclosed a retrolenticular cyclitic membrane and moderately severe (3+) cells and flare anterior chamber reaction. The eye was hypotonous, and B scan showed that the membrane was associated with a retinal detachment. Surgery was performed to repair the retinal detachment and to remove the retrolenticular membrane. Two months before initial examination, the patient had been attacked by a rooster. CONCLUSION: Occult penetrating trauma should be considered in the differential diagnosis of pediatric leukocoria. PMID- 9222250 TI - Choroidal osteoma in an infant. AB - PURPOSE: To report an 8-month-old infant with choroidal osteoma. METHOD: Case report. RESULTS: The patient, who had bilateral yellow stippling at the posterior pole, was followed up for 8 years. Both fundi developed creamy, irregular scalloped lesions. Computed tomography showed a bony plate at the posterior pole bilaterally. CONCLUSION: We believe that this is the youngest patient reported to have choroidal osteoma. PMID- 9222251 TI - Retinal capillary hemangioma in Marshall-Stickler syndrome. AB - PURPOSE: Retinal capillary hemangioma, often associated with von Hippel-Lindau syndrome, is not characteristically seen in other conditions. We report a patient with Marshall-Stickler syndrome who had a retinal capillary hemangioma. METHOD: A 31-year-old man with Marshall-Stickler syndrome was evaluated for a retinal vascular tumor. RESULTS: Evaluation showed a retinal capillary hemangioma of the von Hippel type. Magnetic resonance imaging disclosed no tumors of brain, kidney, or other organs. The retinal tumor was treated with cryotherapy. CONCLUSION: Retinal capillary hemangioma is not pathognomonic of von Hippel-Lindau syndrome, and it may be associated with the Marshall-Stickler syndrome. PMID- 9222252 TI - Ocular neuromyotonia in a patient with cavernous sinus thrombosis secondary to mucormycosis. AB - PURPOSE: To report a case of ocular neuromyotonia occurring after cavernous sinus thrombosis secondary to mucormycosis. METHODS: Case report. We performed serial comprehensive neuro-ophthalmologic examinations. RESULTS: Fifteen months after initial total ophthalmoplegia of the right eye and complete right upper eyelid ptosis, isolated ocular neuromyotonia, characterized by episodic upward jerking movements of the right upper eyelid, was noted. CONCLUSION: Ocular neuromyotonia, which usually manifests in patients with a history of intracranial tumors and cranial radiation, may also be secondary to infectious cavernous sinus thrombosis. PMID- 9222253 TI - Deviated hydroxyapatite orbital implant syndrome. AB - PURPOSE: To describe a patient with a deviated orbital implant after enucleation. METHOD: Case report. RESULT: We examined a postenucleation patient with a medially deviated ("esotropic") implant after peg drilling. Modification of the posterior surface of the prosthesis improved cosmesis in the primary position. CONCLUSIONS: During enucleation surgery, surgeons may use a spherical implant rather than modify the anterior face in patients with preexisting strabismus, with severe trauma to the extraocular muscles, or with risk of implant deviation. Additionally, secondary procedures may alter implant position. Occasionally, patients without obvious preoperative risk factors may have deviation of the modified face of the implant. Flattening the implant creates an asymmetric surface that may lead to a deviated orbital implant. PMID- 9222254 TI - Proptosis as the manifesting sign of Weber-Christian disease. AB - PURPOSE: To identify proptosis as a manifesting sign of Weber-Christian disease (recurrent febrile nodular panniculitis). METHOD: Case report. A 61-year-old man had signs of right proptosis and orbital inflammation that resolved with oral corticosteroid therapy. Orbital inflammation later recurred with associated cutaneous nodules, myalgia, nausea, and malaise. RESULTS: Rheumatologic evaluation and subcutaneous nodule biopsy led to the diagnosis of Weber-Christian disease. The patient required systemic immunosuppressive agents to control the disease. CONCLUSION: This case shows the rare finding of proptosis as the manifesting sign of Weber-Christian disease. PMID- 9222255 TI - Microbiologic factors and visual outcome in the Endophthalmitis Vitrectomy Study. PMID- 9222256 TI - The role of the generalist in the care of the substance-abusing patient. AB - It will be some time before known effective practices on behalf of patients with ATOD problems are integrated into the mainstream of medical care. Ironically, much of the medical literature centers on the medical and psychiatric complications of substance abuse and on physician attitudes regarding treatment of these patients. Rather the focus should be on the ways in which physicians can intervene early and effectively to treat the substance abuse problem itself. Much of this issue serves to correct the imbalance. PMID- 9222257 TI - Screening for alcohol and drug abuse. AB - The purpose of this article is to review screening for substance use disorders in health care settings. The epidemiology of alcohol and other drug abuse is briefly reviewed, followed by a discussion of the principles underlying whether or not screening is warranted. Different screening instruments and strategies are then described. Finally, current recommendations for screening for alcohol and other drug abuse are discussed. PMID- 9222258 TI - Brief interventions with substance-abusing patients. AB - Brief interventions to change alcohol use have proven effective in different primary care settings. Current data show decreases in consumption as well as a decreased utilization of some health resources and decreased mortality. The process of changing drinking or substance abuse behavior requires a series of predictable steps. The key to changing behavior is ambivalence about current behavior. Understanding the stages of behavioral change and assessing the patient's readiness to change are important for effective interventions. Brief interventions include giving the patient feedback about a behavior, clearly recommending a change in behavior, presenting options to achieve this change, checking and responding to the patient's reaction, and providing follow-up care. Effective interventions help develop the patient's sense of motivation and self efficacy for behavioral change. In approaching a patient with a substance abuse problem, using techniques of motivational enhancement is more effective than a confrontational approach. PMID- 9222259 TI - Pharmacotherapies for alcohol abuse. Withdrawal and treatment. AB - Pharmacologic management of alcoholism is only one part of the management of both alcohol dependence and withdrawal, which also includes the provision of a calm, quiet environment; reassurance; ongoing reassessment; attention to fluid and electrolyte disorders; treatment of coexisting addictions and common medical, surgical, and psychiatric comorbidities; and referral for ongoing psychosocial and medical treatment. For further discussion of these topics, the reader is referred to previously published sources. A survey of alcoholism treatment programs revealed that although benzodiazepines were the most commonly used drugs, standardized monitoring of patients' withdrawal severity was not common practice, and a significant minority of clinicians were using a variety of other drugs, some not known to prevent or treat the complications of withdrawal. Treatment should be based on the available evidence (Working Group on Pharmacological Management of Alcohol Withdrawal: American Society of Addiction Medicine Committee on Practice Guidelines: Pharmacological management of alcohol withdrawal: An evidence-based practice guideline. Unpublished draft, 1997). Patients with significant symptoms, patients with complications such as seizures or delirium tremens, and patients at higher risk for complications of alcohol withdrawal should receive benzodiazepines, particularly chlordiazepoxide, diazepam, or lorazepam, because of their safety and documented efficacy in preventing and treating the most serious complications of alcohol withdrawal. These drugs may be dosed on a fixed schedule for a predetermined number of doses on a tapering schedule over several days, or they may be administered by front loading. An alternative approach for selected patients without seizures or acute comorbidity is symptom-triggered therapy, which individualizes treatment and decreases the duration and dose of medication administration. With either of the regimens, patients should have their withdrawal severity monitored until symptoms are resolving. Once withdrawal from alcohol is safely completed, the focus should turn to helping to prevent relapse. Disulfiram may be useful in highly motivated subsets of patients and when compliance-enhancing strategies are used. Naltrexone is useful in the broader population of patients entering treatment for alcohol dependence. These pharmacologic interventions should be given in the context of ongoing psychosocial support. There is substantial evidence that pharmacologic management of alcohol abuse and dependence is effective. As would be predicted from alcohol's myriad cellular effects, no panacea exists for alcoholism. For alcohol withdrawal, however, although treatment regimens have only recently been refined, evidence for effective treatment of symptoms and prevention of complications with benzodiazepines has been available for decades. Within the last decade, effective treatments, including naltrexone, have been shown to reduce alcohol intake in alcohol-dependent persons. Given the prevalence and cost of alcohol-related problems, all effective therapies (including pharmacologic treatments) should be considered to treat alcohol abuse and dependence. PMID- 9222260 TI - Pharmacotherapy for opioid and cocaine abuse. AB - The need for continued development of medications to address opioid and cocaine addiction is unequivocal. Methadone maintenance, despite its limitations, remains the best-established pharmacologic treatment for opioid dependence. Continued participation in methadone programs is associated with decreased risk of acquiring HIV infection. Clonidine alone or combined with naltrexone may be used for opioid detoxification in the office practice. At the present time, no proven pharmacologic therapy for cocaine addiction exists. PMID- 9222261 TI - Nonpharmacologic approaches to substance abuse treatment. AB - Familiarity with nonpharmacologic approaches to substance abuse treatment is critical for medical practitioners to act effectively to prevent the progression of substance use to medically harmful use, abuse, or dependence; to identify patients with substance use disorders and motivation behavioral changes; and to maximize the likelihood of successful treatment. At their most basic level, these nonpharmacologic approaches involve components of practice that are requisite to the successful management of any medical disorder: fostering an empathic, supportive relationship; routinely evaluating the system or problem area; providing accurate medical information about diagnosis, natural history, and treatment; and following up on identified problems to improve compliance, evaluate the impact of treatment, and modify treatment as indicated. Because of the nature of substance use disorders, their impact on multiple areas of functioning, and the conditioned craving that occurs following repeated substance use, nonpharmacologic treatments can improve outcome, even when effective pharmacologic treatments are also employed. Treatment of nicotine dependence provides a useful example. Physician advice to stop smoking substantially increases the likelihood of smoking cessation and long-term abstinence. Combined with physician advice, nicotine replacement therapies, using nicotine gum or transdermal preparations, approximately double the rate of long-term abstinence, compared with physician advice alone. Providing behavioral treatment in addition to physician advice and nicotine replacement treatment leads to the highest rates of sustained abstinence, significantly higher than advice alone or rates associated with nicotine replacement alone. Nonpharmacologic treatments complement pharmacologic approaches often by addressing different target symptom and problem areas. In the case of nicotine dependence, nicotine replacement ameliorates withdrawal symptoms and craving associated with withdrawal. Behavioral treatment improves outcome by focusing on cue-evoked craving and developing effective long-term strategies to avoid or cope with craving and other cues. As discussed in this article, brief motivation approaches are particularly well suited for general medical practice settings. These approaches have been evaluated most extensively and shown to be most effective in preventing the progression of heavy drinking to problem drinking and alcohol dependence. Following a thorough evaluation of a patient's drinking habits, providing advice about sensible and safe drinking to patients identified as heavy drinkers leads to meaningful reductions in drinking. For patients who have developed problems of abuse or dependence, motivation approaches can be used to foster an interest and commitment to stop use and accept a referral to treatment. This article also provides an overview of the major psychosocial approaches used in more intensive specialty treatment of patients with substance use disorders. Familiarity with these approaches is essential for clinicians in general medical settings to facilitate referral of patients and monitor and improve the efficacy of treatments provided to patients. Medical practitioners must be able to educate patients about the need for more intensive specialty treatment and about what treatment entails. Medical practitioners must also be able to engage in informed discussions with substance abuse treatment specialists about the specific treatment recommendations made for a patient and the rationale for them. Medical practitioners who are informed about the treatment plans, rationale for treatment, and patient progress can play critical roles in encouraging patients to persist with the often difficult process of treatment. PMID- 9222262 TI - Treatment matching. Theoretic basis and practical implications. AB - An extensive knowledge base supports the development of treatment matching methods for alcohol and other drug abuse. Many matching variables have been identified that relate to specific modalities of treatment and to specific levels of care. Physicians and other providers can use many putative matching variables to address patients' substance use problems on a highly individualized basis. These variables include demographic factors (age, gender, culture), typology and severity (age of onset; severity of intoxication; withdrawal; quantity, recency, frequency of substance use), intrapersonal characteristics (psychiatric diagnosis, cognitive function, self-efficacy, stage of change), and interpersonal function (social stability). The evolution of formal criteria for patient placement such as the ASAM criteria is a beneficial, adaptive process that is underway in numerous states, managed care entities, professional provider societies, and provider groups. Currently, matching approaches rely more heavily on consensus recommendations than on empiric matching data. The technology for conducting psychosocial treatment matching studies is rapidly increasing in sophistication. Although predictive validity has not yet been fully demonstrated on a large, multisite basis, the national research portfolio on treatment matching is expanding in size and complexity. This is an essential public health need, given dramatic cost pressures, if addiction services are to continue to grow in quality and availability. PMID- 9222263 TI - Prescription drug abuse. A question of balance. AB - The current American medical practice paradox of concomitant overprescribing and underprescribing of controlled drugs is within the power of physicians to correct. It requires actively seeking education in traditionally neglected areas and avoiding prescribing controlled drugs to patients with either substance abuse histories or vague clinical indications. Attempts to limit prescribing to short therapeutic time courses, refusal to prescribe if pushed, and careful chart documentation practices are important. By increasing knowledge about chemical dependence and about chemically dependent patients' abnormal relationships with scheduled drugs, the current clinical reality can be reversed. This reversal will result in marked decreases in prescribing to the minority of patients (those with chemical dependence) and increases in prescribing to the majority of patients who are currently often undertreated. PMID- 9222264 TI - Women and substance abuse. AB - Chemically dependent women face special problems. This article reviews the epidemiology, screening, clinical consequences, and treatment of substance abusing women. Alcohol, opiate, and cocaine abuse are often linked in women, and the individual and overlapping effects of these drugs are described. Gender difference also are highlighted. PMID- 9222265 TI - Geriatric substance use disorders. AB - AUDs are increasingly recognized as common problems among older adults. The magnitude of this problem is likely to increase over ensuing decades as baby boomers reach retirement age with drinking habits that are significantly different from current cohorts of older adults. Barriers to detection are numerous and include nonspecificity of alcohol-related presentations, patient denial, and clinicians' unwillingness to recognize that patients can and do develop alcohol problems in later life. Despite the limitations of current screening and diagnostic instruments, the authors recommend use of the CAGE as a formal screening tool for older patients because of its brevity, demonstrated efficacy, and convenience. In patients who answer affirmatively to any CAGE question, diagnostic certainty can be increased by use of follow-up questions or referral to an alcohol treatment specialist. Referral of patients with established alcohol abuse or dependence is essential for definitive treatment, and successful outcomes can be expected and are gratifying once achieved. In patients with less severe AUDs, brief interventions with frequent follow-up are recommended. Age-specific screening and diagnostic instruments for older AUD patients, once fully developed and validated, will facilitate identification. Much less is known about other substance use disorders in older adults. Psychoactive drug use is not uncommon in this patient population and may result in adverse health outcomes. Treatment interventions proposed for AUDs are advocated for older adults found to have other substance use disorders as well and are likely to yield improved outcomes. Future investigations that better define the epidemiology, detection, and treatment of other substance use disorders in older populations are clearly warranted at this time. PMID- 9222266 TI - Dual diagnosis in primary care. Detecting and treating both the addiction and mental illness. AB - The initial phase of treatment includes engaging the patient in a discussion about the doctor's concerns and providing patients with information about the problems as well as the possibility of change. Treatment of dual disorders often requires a heightened awareness of the consequences of the problem and the development of a realistic plan for change. The treatment plan must attempt to evaluate and treat the addiction and the psychiatric and medical illnesses. PMID- 9222267 TI - Physician impairment by substance abuse. AB - Physician impairment by substance abuse represents a significant challenge to physicians, patients, and society as a whole. Although data is sparse, the prevalence of alcohol and illicit drug abuse among physicians is probably similar to that of the general population, while abuse of prescription drugs may be more prevalent. From a medicolegal standpoint, these issues are managed mostly at the state level and substance abuse is of increasing interest to credentialling organizations such as hospitals and managed care organizations. A variety of concrete steps can be taken to identify physicians with substance abuse problems and treatment approaches have been designed specifically for impaired physicians. With improved attention to the problem of physician impairment by substance abuse, the well-being of both physicians and their patients can be enhanced. PMID- 9222268 TI - Managed care of substance abuse disorders. Implications for generalist physicians. AB - Given the rapidly changing landscape of both private and public health plan programs, it is foolhardy to suggest that a definitive characterization of managed care and substance abuse issues can be made. Among other topics, this article discusses goals of managed care organizations, managed care models, and physicians' roles in managed care. PMID- 9222269 TI - The changing standard of care in Parkinson's disease: current concepts and controversies. PMID- 9222270 TI - Contemporary approaches to the pharmacotherapeutic management of Parkinson's disease: an overview. AB - A number of unresolved issues complicate the effective management of patients with Parkinson's disease (PD). Chief among these is the role of neuroprotective versus symptomatic pharmacologic interventions. Until the etiology of PD is further defined, consensus on appropriate management of this illness is unlikely. Clinicians may best serve their patients by taking a pragmatic approach to the treatment of PD that utilizes potentially beneficial interventions in eligible patients. Such an approach would incorporate possible neuroprotective (e.g., selegiline, dopamine agonists, sustained-release levodopa) and dopamine-sparing (e.g., combination levodopa/dopamine agonist therapy) strategies whenever possible while retaining adequate symptomatic control. PMID- 9222271 TI - Issues in the early diagnosis of Parkinson's disease. AB - The clinical diagnosis of Parkinson's disease (PD) is most difficult early in the disease when the signs and symptoms are most subtle. The differential diagnosis of PD includes a number of movement disorders with similar symptomatology (e.g., essential tremor, multiple system atrophy, vascular parkinsonism). In most published studies of PD, the disease is diagnosed simply by the presence of two of the three cardinal motor signs-tremor, rigidity, and bradykinesia-or by the presence of three of the four motor signs: tremor, rigidity, bradykinesia, and postural instability. However, there is an obvious need for better diagnostic criteria. Until discrete biologic markers are developed, the use of exclusion criteria may improve the accuracy of the presumptive diagnosis of PD. PMID- 9222272 TI - Attempts to obtain neuroprotection in Parkinson's disease. AB - It is suggested that oxidant stress is a contributing factor in the pathogenesis of Parkinson's disease (PD). Oxidant stress may contribute to cell death in PD because oxidative metabolism of dopamine has the potential to yield highly reactive and cytotoxic free radicals. Evidence for this hypothesis includes: (1) increased dopamine turnover with increased hydrogen peroxide formation; (2) decreased glutathione availability; and (3) increased reactive iron in the brains of patients with PD. Antioxidant therapies might be neuroprotective and could slow the clinical progression of the disease whereas metabolites of levodopa therapy may accelerate the rate of neuronal degeneration. Laboratory studies demonstrate that both selegiline and dopamine agonists can provide neuroprotective benefits. Selegiline-treated patients require less levodopa and have a delay in the progression of parkinsonian signs and symptoms. Dopamine agonists provide antiparkinson benefits and also diminish the need for levodopa. PMID- 9222273 TI - The role of dopamine agonists in early Parkinson's disease. AB - Recent evidence from clinical studies suggests an expanded role for dopamine agonists as initial dopaminergic monotherapy in the treatment of Parkinson's disease (PD). The rationale for the use of dopamine agonist monotherapy in early disease is to delay the initiation of levodopa or to decrease the total exposure to levodopa, thereby reducing the motor complications of long-term levodopa therapy. Dopamine agonists, when used alone, rarely promote the development of dyskinesias and motor fluctuations that complicate levodopa treatment. Theoretically, there is potential for a neuroprotective effect by decreasing the oxidative breakdown of dopamine and free radical generation. Because they act on postsynaptic dopamine receptors of the striatum, dopamine agonists act independent of the synthetic dopaminergic enzyme system and are not dependent on degenerating presynaptic neurons in the substantia nigra. This article will review the traditional role of dopamine agonists and will focus on emerging strategies for the treatment of PD, including early monotherapy with dopamine agonists and early combination therapy with dopamine agonists and levodopa. PMID- 9222274 TI - Managing the late complications of Parkinson's disease. AB - Most patients with Parkinson's disease (PD) receiving chronic levodopa therapy eventually manifest one or more motor response complications, including "wearing off" phenomena and "on-off" phenomena. Additionally, as the disease progresses, motor, neurologic, and neuropsychiatric complications increase and may include freezing spells, falls, dementia, depression, and psychosis. The management of patients with advanced PD presents a special clinical challenge because patients may experience an enhanced sensitivity to small changes in plasma levodopa levels and because they may suffer adverse reactions to antiparkinsonian drugs. Management of advanced PD is directed toward decreasing the dose of the offending drug while raising the dose of an alternative drug, with the goal of maintaining symptom control. In this article, the spectrum of late complications experienced by patients with advanced PD and their management are discussed. PMID- 9222275 TI - Pharmacologic profile of ropinirole: a nonergoline dopamine agonist. AB - The use of dopamine agonists as monotherapy or in combination with levodopa in the treatment of Parkinson's disease (PD) allows for reduction or limitation of the levodopa dose, potentially delaying the onset or reducing the severity of late motor complications. Ropinirole is a new nonergoline dopamine agonist that binds specifically to D2-like receptors with a selectivity similar to that of dopamine (D3 > D2 > D4). The chemical structure of ropinirole has the potential to maintain a structure-activity relationship similar to that of dopamine and other effective dopamine agonists without producing ergot-related adverse effects. Ropinirole has demonstrated efficacy in two standard preclinical models of PD and has shown a very low propensity to induce dyskinesia in these studies. This latter property is of potential clinical importance for pharmacotherapy of early PD. This article will present the importance of pharmacologic specificity of dopamine agonists along with the basic pharmacologic characteristics of ropinirole that may contribute to its efficacy in the treatment of PD. PMID- 9222276 TI - Human retinoblastoma: in vitro differentiation and immunoglobulin superfamily antigen modulation by retinoic acid. AB - Suspension and attachment cultures of Y79 human retinoblastoma cells were treated with all-trans retinoic acid (RA) for up to 10 days to assess its effect on growth and cell-surface expression of immunoglobulin superfamily antigens MHC class I and class II, ICAM-1, NCAM and Thy1. RA up to 10 microM induced growth inhibition, and marked morphological differentiation with extension of prominent processes resembling neurites was seen in attachment cultures. However, above 10 microM RA produced extensive cell death. We also observed increased cell-surface expression of MHC class I, ICAM-1, NCAM and Thy1 on Y79 cells treated with 10 microM over 10 days; constitutive MHC class II expression was not apparent, nor did RA treatment appear to induce Y79 cells to express MHC class immunoreactivity. The up-modulation of cell-adhesion molecules (NCAM, ICAM-1 and Thy1) and immune recognition molecules (NCAM, ICAM-1 and MHC class I), associated with reduced growth and tumour cell differentiation, suggests that RA may have a potential role in regulating the growth and development of retinoblastoma tumours. PMID- 9222277 TI - Active immunization of metastatic melanoma patients with interleukin-2-transduced allogeneic melanoma cells: evaluation of efficacy and tolerability. AB - From January 1994 to July 1996 we immunized metastatic melanoma patients with HLA A2-compatible, interleukin-2 (IL-2)-secreting, immunogenic melanoma lines in an attempt to induce a systemic reaction that might also affect distant melanoma lesions. Twelve patients (6 male and 6 female) aged from 28 to 72 years, affected with visceral and/or subcutaneous (s.c.) melanoma metastases, were treated. Two different HLA-A2+ melanoma lines were transduced with the human IL-2 gene (14932/IL-2 and 1B6/IL-2) and used as vaccine. Two groups of 4 patients each were injected s.c. with 5 x 10(7) and 15 x 10(7) irradiated 14932/IL-2 melanoma cells respectively, whereas a third group received 5 x 10(7) cells of the second line (1B6/IL-2). All patients received the vaccine on days 1, 13, 26; if no progression was evident, further immunizations were administered at monthly intervals. All patients were assessable for clinical response after at least three injections of the vaccine. In 4 cases a stabilization of disease lasting from 2 to 6 months was observed: in 2 of them a mixed type of response to treatment was noted with simultaneous evidence of regressing and non-responding lesions in the same patients. No signs of clinical response were found in the remaining patients. Nine patients died of disease between 3 and 24 months after the onset of therapy, whereas 3 were alive 3 months after the end of therapy. The local and systemic side-effects of treatment were mild. These results indicate that vaccination with cells bearing the appropriate antigens and releasing IL-2 locally can produce weak clinical responses, but also indicate that better results may be achieved through modifications of the vaccine, the schedule of immunization and/or a more appropriate selection of patients. PMID- 9222278 TI - Adoptive transfer of cytotoxic T lymphocytes induced by CD86-transfected tumor cells suppresses multi-organ metastases of C1300 neuroblastoma in mice. AB - In this study, we examined the therapeutic antitumor effect of cytotoxic T lymphocytes (CTL) generated against CD86-transfected mouse neuroblastoma C1300. We first generated the transfectant, CD86 + C1300, expressing a high level of mouse CD86 on the cell surface. While CD86 + C1300 cells were rejected in syngeneic A/J mice when inoculated subcutaneously, neither vaccination nor any therapeutic antitumor effect was obtained, implying that C1300 may be a poorly immunogenic tumor. However, in vitro stimulation of splenocytes from either C1300 bearing or CD86 + C1300-rejecting mice with CD86 + C1300 cells resulted in remarkable CTL activity against C1300 cells. The CTL activity induced by CD86 + C1300 was mediated by T cell receptor/CD3 and CD8 and was further enhanced by the addition of interleukin-2. Intravenous inoculation of C1300 cells led to multiple organ metastases including the liver, lung, kidney, ovary, lymph node and bone marrow. To examine the therapeutic effect of CTL in this metastasis model, CTL induced by parental or CD86 + C1300 cells were administrated into C1300-bearing mice. Adoptive transfer of CD86 + C1300-induced CTL resulted in marked elimination of multi-organ metastases and prolonged survival in almost all mice, 70% of which survived indefinitely. These results indicate that adoptive transfer of CTL induced by CD86-transfected tumor cells in vitro would be effective and useful for tumor immunotherapy against poorly immunogenic tumors. PMID- 9222279 TI - Relation between HPV-16 serology and clinico-pathological data in cervical carcinoma patients: prognostic value of anti-E6 and/or anti-E7 antibodies. AB - To investigate the clinical significance of the enhanced sensitivity of antibody detection by radio immunoprecipitation assays (RIPA), using in vitro translated HPV-16 E6 and E7 proteins, over synthetic-peptide enzyme-linked immunosorbent assay (ELISA), RIPA for HPV-16 E6 and E7 were performed. The results obtained with E6 and E7 RIPA were related to clinico-pathological data from cervical carcinoma patients positive for HPV type 16 DNA in their primary tumour. The data obtained with E6 and E7 RIPA were then compared to the results obtained using the E7/6-35 synthetic-peptide ELISA. The prevalence of antibodies to E6, E7, E6 and/or E7 and E6 and E7, as determined by RIPA, was significantly higher in cervical cancer patients than in both controls and cervical intraepithelial neoplasia patients. Odds ratios, calculated for cervical carcinoma patients versus controls, ranged from 7.4 to 15.4. Antibodies to E6 and/or E7 were largely restricted to patients with HPV DNA in their primary tumour. Analysis of the relation between prevalence of antibodies to E6 and E7 and clinico-pathological parameters was limited to 85 patients positive for HPV-16 DNA. The strongest relation with clinico-pathological data, such as lesion size, lymph node involvement, and prognosis, was found for E7 synthetic-peptide ELISA, whereas E6 and E7 RIPA did not reach significance. The significance of these findings is discussed. PMID- 9222280 TI - Synergistic effects of Actinobacillus suis cells in combination with 5 fluorouracil on mice bearing murine L1210 leukemia cells. AB - The mean survival rates of female BDF1 mice transplanted intravenously (i.v.) with murine L1210 leukemia cells were significantly prolonged by intraperitoneal (i.p.) pretreatment (before i.v. transplantation) or by i.p. pre- and post treatment (before and after the i.v. transplantation) with heat-killed Actinobacillus suis cells ATCC 15557 (AS 15557) alone, as compared with untreated L1210-leukemia-cell-bearing control mice. However, significant prolongation of the mean survival rates was not elicited by the i.p. post-treatment with AS 15557 alone. When 5-fluorouracil (5-FU) was applied i.p. to mice receiving post treatment with AS 15557 alone, the mean survival rates of the L1210-leukemia-cell bearing mice were significantly prolonged. The antileukemic action of AS 15557, alone or in combination with 5-FU, against L1210 leukemia was superior to that of a streptococcal preparation (OK-432) and was almost the same as of bacillus Calmette-Guerin with or without 5-FU. The results suggest the possibility that the synergism of AS 15557 in combination with 5-FU may be dependent on the relationship between the indirect immunological function of AS 15557 and the direct cytotoxic action of 5-FU on L1210 leukemia cells. PMID- 9222281 TI - Priming of the antitumor response promotes efficacy of interleukin-2 therapy. AB - We have studied the effect of active specific immunization (ASI) on the antitumor response induced by locoregional, low-dose interleukin-2 (IL-2) therapy. On day 0, mice were injected i.p. with viable, syngeneic tumor cells and with irradiated tumor cells (ASI). Low-dose IL-2 treatment was given i.p. for 5 consecutive days. ASI led to extended survival in two out of seven models tested. In these two models, enhanced efficacy was observed when both ASI and IL-2 were administered. In the five models in which ASI had no therapeutic value, ASI + IL-2 treatment was no more effective than IL-2 therapy. In the SL2 lymphoma model, use of ASI prior to IL-2 therapy given as early as days 1-5 led to at least 60% cure, whereas IL-2 therapy without ASI was only effective when administered after day 9. In the P815 mastocytoma model, however, ASI, IL-2, and the combination caused negative (suppressive) effects when administered on days 6-10. IL-2 administered on days 6-10 was therapeutically effective in this model when mice were treated with cyclophosphamide on day 6. In both the SL2 and the P815 tumor models, cured mice were specifically immune. The positive and negative effects observed were not due to the increased number of cells injected (non-specific inflammation) nor to possible antigenic alteration of the ASI cells by irradiation, as ASI with fragmented tumor cells was also effective in inducing synergy. Investigations into the underlying mechanism indicated that CD4+ cells play an important role. In total, the results indicate that ASI may be a good supplement to locoregional IL-2 treatment if care is taken to alleviate immunosuppressive activities. PMID- 9222282 TI - Targeting an anti-viral CD8+ T cell response to a growing tumor facilitates its rejection. AB - We demonstrate in a murine model that targeting an anti-viral T cell response to a growing tumor facilitates priming of a tumor-associated antigen (TAA)-specific, rejecting T cell response. Murine P815 mastocytoma cells grow aggressively in a syngeneic host. Transfected P815/S cells (expressing the hepatitis B surface antigen, HBsAg) also grow as subcutaneous tumors, but occasional 'spontaneous' rejections after transient growth are observed. Growth of P815/S tumors (but not of P815 tumors) is efficiently suppressed by a CD8+ cytotoxic T lymphocyte (CTL) dependent immune mechanism in mice primed to HBsAg by DNA-immunization. In hosts immunized against HBsAg by DNA vaccination, HBsAg-specific CTL are generated. This specific CTL reactivity was targeted to s.c.-growing P815 tumors by intra tumor injections of either HBsAg-encoding plasmid DNA or viable P815/S cells; this treatment led to tumor rejection in 70-80% of the tumor-bearing animals. All rejecting animals showed a CD8+ CTL-dependent resistance to subsequent challenges by native, non-transfected P815 tumors. Targeting an established anti-viral ('strong') CTL response to a growing tumor hence is an efficient strategy to facilitate priming of a rejecting CTL response against ('weak') TAA in this system. PMID- 9222283 TI - Analysis of Pmel17/gp100 expression in primary human tissue specimens: implications for melanoma immuno- and gene-therapy. AB - Pmel17/gp100-encoded tumor-associated antigens are recognized by cytotoxic T lymphocytes in melanoma patients and may represent attractive target antigens for immuno- and gene-therapeutic strategies. An important prerequisite for identification and monitoring of melanoma patients that could potentially benefit from Pmel17/gp100-based immuno- and gene-therapies is the detailed knowledge of Pmel17/gp100 expression in vivo. Immunophenotyping is considerably hampered by the different immunoreactivities of Pmel17/gp100-reactive antibodies. Therefore, we analyzed an extended series of different primary normal and malignant human tumor specimens for Pmel17/gp100 expression at the mRNA level. Transcripts were detectable in all malignant melanoma tissue specimens representing all stages of tumor progression, with significant levels even in early and amelanotic melanoma lesions. In contrast, normal melanocytes exhibited significantly less Pmel17/gp100 mRNA in vivo, as determined by comparative in situ hybridization. Tissue specimens from the retina and substantia nigra also contained Pmel17/gp100 mRNA, whereas other normal and malignant human tissues were negative. As determined by comparative in situ hybridisation and HMB-45 immunostaining, even tumor tissue lacking Pmel17/gp100 immunoreactivity contained Pmel17/gp100 transcripts. Our results indicate a melanocytic-cell-lineage-restricted expression of Pmel17/gp100 with significant transcript levels in all stages of melanoma progression, including early and amelanotic melanoma lesions, and a significantly differential expression between melanoma cells and normal melanocytes in vivo. Owing to its higher sensitivity, phenotyping of individual tumor specimens by mRNA expression analysis seems to be more valuable than phenotyping by immunostaining. PMID- 9222284 TI - In vitro improvement of chlorambucil-induced cytotoxicity by deflazacort and 6 methylprednisolone in B-cell chronic lymphocytic leukaemia. AB - We examined whether CLL cell chemosensitivity to in vitro exposure to chlorambucil (CLB) might be improved by the presence of deflazacort (DFZ) in comparison to 6-methylprednisolone (PDN). The PDN lethal dose (LD)50 values were low in 5 samples, intermediate in 4 and high in 7. Low, intermediate and high DFZ LD50 values were detected in 3, 2 and 11 samples, respectively. The CBL-LD50 mean values were significantly reduced at all PDN and at the 4 highest DFZ concentrations. However, a dose-response effect was seen only in the DFZ group. Both CLB-DFZ and CLB-PDN interactions were analysed in 16 samples at 25 different dose-combinations, resulting in 400 comparisons between expected and observed leukaemic cell survival (LCS) values for each group. In particular, 45.75% and 40% dose combinations were synergistic in CLB-DFZ and CLB-PDN groups, respectively. A relatively higher number of antagonistic interactions were observed among CLB-PDN dose combinations, while analogous number of additive interactions were detected. At concentrations of CLB x1 microgram/ml the phenomenon of synergism, regardless of the steroid concentration, did occur more frequently. On the other hand, a more elevated number of antagonistic interactions were counted at CLB 100 micrograms/ ml. In conclusion, both DFZ and PDN synergize in vitro with CLB, especially at low concentrations of the alkylating agent. PMID- 9222285 TI - Evidence for a NO synthase in porcine platelets which is stimulated during activation/aggregation. AB - We tried to characterize the porcine platelet nitric oxide (NO) synthase and its L-arginine (L-arg)/NO metabolism. Using RT-PCR we could show a constitutive endothelial NOS (ecNOS) and an inducible NOS (iNOS) similar mRNA in platelets. The NOS protein could be evidenced by an ecNOS specific antibody which also bound in platelets. This finding could be confirmed by Western blot showing an ecNOS in the membrane but not the cytosolic fraction; iNOS protein could not be detected. Using NADPH-diaphorase staining we could show NO synthase in preactivated platelets but not in resting platelets, indicating that the platelet NOS may be activated during platelet activation/aggregation. Porcine L-arg plasma levels (9.31 x 10(-5) mol/l +/- 10%) could be shown to be in the same range as human plasma levels. Moreover, we could show that the NO precursor L-arg and hydroxy-L arginine (OHarg) concentration dependently inhibited collagen induced platelet aggregation. Summarizing these results confirm the existence of and further characterize porcine platelet NO synthases. PMID- 9222286 TI - Clinical results of recombinant erythropoietin in transfusion-dependent patients with refractory multiple myeloma: role of cytokines and monitoring of erythropoiesis. AB - Recombinant erythropoietin (r-EPO) was administered to 37 patients with advanced, transfusion-dependent and chemo-resistant multiple myeloma (MM), at the fixed dose of 10,000/U s.c., 3 times a week, for 2 months. Thirteen patients (35.1%) achieved a significant response in terms of complete abolition of red cell transfusions. Factors significantly predictive of response were: a) inappropriate production of endogenous EPO, as expressed by a reduced observed/predicted ratio; b) presence of a consistent number of circulating erythroid precursors BFU-E; c) low serum levels of tumor necrosis factor (TNF) and interleukin-1 (IL-1), cytokines with inhibitory activity on erythropoiesis; d) a single line of previously received chemotherapy. Renal failure, bone marrow plasma cell infiltration, serum levels of IL-6 and other main clinical and laboratory parameters did not affect significantly the response to r-EPO. High fluorescence reticulocytes (HFR) and soluble transferrin receptor (sTfR) values were useful to detect an early stimulation of erythropoiesis in responders, while a high percentage of circulating hypochromic erythrocytes (HE), as assessed by an automated counter, identified those patients developing functional iron deficiency during r-EPO treatment. We conclude that about one-third of severely anemic patients with advanced MM, unresponsive to chemotherapy, may benefit by r EPO therapy. The clinical management of these patients can be accomplished using non-invasive parameters, such as sTfR, HFR and HE. PMID- 9222287 TI - Maximal gamma-globin expression in the compound heterozygous state for -175G gamma HPFH and beta degree 39 nonsense thalassaemia: a case study. AB - The -175 (T-->C) G gamma hereditary persistence of fetal haemoglobin is a very rare promoter mutation occurring in Caucasians as well as in African-Americans. Heterozygotes for this non-deletional HPFH show 20% HbF, mostly of G gamma type. We describe here a healthy Sardinian man who coinherited -175 (T-->C) G gamma HPFH with the beta-thalassaemia codon 39 nonsense mutation in trans; he showed 64% HbF, 100% of G gamma type. Although the beta-globin haplotype pattern (II/II) was indicative of the presence of the A gamma T allele on both chromosomes, the A gamma T expression was undetectable by HPLC even in red cell populations separated by age. The proband was, moreover, homozygous for the -4 bp deletion at position -225 to -222 of A gamma promoter which has recently been associated with decreased A gamma T globin expression. These findings suggest that this maximal overexpression of G gamma-globin probably reflects intensified stimulation of the mutated G gamma promoter in this hitherto undescribed genetic condition. PMID- 9222288 TI - Acute toxicity and effectiveness of idarubicin in childhood acute lymphoblastic leukemia. AB - Anthracyclines have become important components of multi-agent remission induction and continuation therapy of acute lymphoblastic leukemia (ALL). New anthracycline derivatives are being investigated in an attempt to shift the balance of side effects and antileukemic potency. To evaluate the toxicity and efficacy of idarubicin (IDA) in childhood ALL, a prospective multicenter phase-II study was performed. A total of 51 children with prognostically poor recurrences of ALL were enrolled, all of whom had been exposed to anthracyclines during front line treatment. A single 48-h continuous infusion of IDA at 24 mg/m2 was started on the first day of salvage treatment without concomitant systemic cytostatic agents. The response was assessed by reduction of leukemic blasts in the bone marrow and other compartments 2 wk later. IDA monotherapy caused complete and partial remissions in 5 and 20 patients, respectively (49%). Delays of treatment with subsequent polychemotherapy courses were frequent and mainly caused by prolonged intervals of myelosuppression and high rates of systemic infection. Non hematological toxicities including acute cardiac reactions were transient and moderate. Our findings suggest that IDA is an effective drug for remission induction in children with ALL, with acute hematological toxicity being dose limiting. PMID- 9222289 TI - Circadian cell cycle variations of erythro- and myelopoiesis in humans. AB - By use of a multiparameter flow cytometric method with specific surface markers, circadian (24-h) variations in cell cycle distribution have been studied in 19 healthy male volunteers by sampling bone marrow (BM) every 4-5 h during 24-h periods. Admixture of peripheral blood during the sampling was specifically adjusted for, and the fractions of cells in DNA synthetic phase were measured for different hemopoietic cell lineages. Significant circadian variations in DNA S phase were demonstrated both in myelo- and erythropoiesis of the human BM, with 75% (myeloid) and 80% (erythroid) of the volunteers showing highest activity (values) of DNA S-phase during the day and lowest activity (values) between midnight and 04:00 h. A temporal relationship in the circadian variation of S phase and G2/M-phase was demonstrated between the myeloid and erythroid cell lineages. The highest fractions of S-phase cells were found in erythropoiesis, while the highest circadian stage dependent variation was found in myelopoiesis. The existence of a similar phasing in DNA synthetic activity for myelopoietic and erythropoietic cells in the human bone marrow indicates that the circadian rhythmicity of hemopoiesis may be caused by a common regulatory mechanism. These findings may be relevant with regard to optimizing the use of cytotoxic drugs and hemopoietic growth factors by taking into consideration the intrinsic (endogenous) circadian variation in proliferative activity of human BM subpopulations. PMID- 9222290 TI - Dominant-negative mutations of the Wilms' tumour predisposing gene (WT1) are infrequent in CML blast crisis and de novo acute leukaemia. AB - To determine if mutations of the Wilms' tumor predisposing gene (WT1) are associated with haematological malignancies, we have investigated 65 cases of acute leukaemia, including 39 patients in blast crisis of chronic myeloid leukaemia (CML), by amplification of WT1 exons 7, 8 and 9 followed by single strand conformation polymorphism analysis. WT1 transcripts were detected by RT PCR in all samples. An exon 7 silent polymorphism (A-->G; Arg 313) was identified in 17 individuals, 5 of whom were homozygous, but no other lesions were found. In 1 sample from a patient with acute lymphoblastic leukaemia a smaller size transcript missing exon 9 was detected; a similar abnormality has been described previously in a patient with Wilms' tumour and the resultant protein shown to act in a dominant-negative manner. No mutations of the exon 9 donor or acceptor splice sites were found in this patient and the basis of the abnormal transcript remains obscure. We conclude that dominant-negative mutations of the zinc finger region of the WT1 gene are uncommon in CML blast crisis. Abnormalities of this gene may, however, contribute to a small proportion of cases of de novo acute leukaemia. PMID- 9222292 TI - The influence of tumour necrosis factor-alpha on phagocytosis of human erythrocytes by autologous monocytes/macrophages. PMID- 9222291 TI - Platelet membrane fluidity and intraplatelet Ca2+ mobilization are affected in uraemia. AB - In present investigations, platelet membrane fluidity and intraplatelet Ca2+ mobilization were analysed in uraemic platelets by fluorescence techniques. Thirteen non-dialyzed uraemic patients and 16 control subjects were examined. Anisotropy of DPH-probe, measured at 37 degrees C, was significantly higher in control (0.2236 +/- 0.0050) than in uraemic platelets (0.1969 +/- 0.0082; p < 0.01). There was no difference between control (109.8 +/- 6.0 nM) and uraemic platelets (100.0 +/- 7.3 nM) when the basal [Ca2+]i in resting platelets was determined. Activation of platelets by ADP (12.5 microM) or by thrombin (0.1 U/ml) resulted in an increase in [Ca2+]i. It was significantly higher (p* < 0.003 for ADP and p* < 0.009 for thrombin, respectively) in control platelets (383.6 +/ 56.3 nM and 2031.0 +/- 298.8 nM, respectively) than in uraemic ones (191.0 +/- 21.3 nM and 838.7 +/- 144.1 nM, respectively). The amount of released Ca2+ was higher in control platelets activated by both ADP and thrombin (157.6 +/- 21.4 nM and 409.3 +/- 71.0 nM, respectively) than in uraemic platelets (76.7 +/- 15.7 nM and 203.0 +/- 29.3 nM, respectively) and the differences were significant (p < 0.01 and p* < 0.01, respectively). These results indicate an abnormal intracellular Ca2+ mobilization in uraemic platelets. Both increased membrane fluidity and decreased Ca2+ mobilization should be considered as a possible reason of reduced fibrinogen receptor exposure on uraemic platelets. PMID- 9222293 TI - Immunization against platelet glycoprotein IIb-IIIa in Glanzmann's thrombasthenia. PMID- 9222294 TI - Reticulocyte recovery is faster in allogeneic and autologous peripheral blood stem cell transplantation than in bone marrow transplantation. PMID- 9222295 TI - Stimulatory effect of reconstituted basement membrane components (matrigel) on the colony formation of a panel of human lung cancer cell lines in soft agar. AB - Lung cancers have been distinguished into small-cell lung cancer (SCLC) and non small cell-lung cancer (NSCLC) types on the basis of their clinical behaviors and their responses to treatment. Moreover, growth of most SCLC cell lines in liquid culture medium is nonadherent, while that of most NSCLC cell lines is adherent. In this study, we examined the effect of matrigel (reconstituted basement membrane components), which is known to have growth-stimulatory activity on various human tumor cell lines in immunodeficient mice, on soft-agar colony formation of a panel of SCLC and NSCLC cell lines to clarify its mechanism of growth stimulation of cancer cells. Matrigel enhanced colony formation of all 9 NSCLC cell lines and 4 of 9 SCLC cell lines. There was a statistically significant difference (P < 0.01) between colony formations with and without matrigel of NSCLC cell lines, but not for SCLC cell lines. In liquid culture medium, all 9 NSCLC lines and 3 of 9 SCLC lines adhered to plastic dishes, whereas the other SCLC lines did not. Matrigel enhanced colony formation of all 3 adherent-type SCLC lines and 1 of 6 nonadherent-type NSCLC lines. Matrigel enhanced colony formation of both of 2 adherent-type non-lung cancer cell lines and 1 of 2 nonadherent-type leukemia cell lines. Neither transforming growth factor beta, collagen type IV, fibronectin, nor laminin, which are components of matrigel, enhanced colony formation of an NSCLC cell line in soft agar. The increase in the colony number of the NSCLC cell line by matrigel was abrogated by the protein kinase inhibitors staurosporine and UCN-01. PMID- 9222296 TI - Invasive potentials of gastric carcinoma cell lines: role of alpha 2 and alpha 6 integrins in invasion. AB - The potentials of the two major histological types of gastric carcinoma to invade through extracellular matrices were studied with cell lines. We found that the invasive potential of intestinal-type carcinoma cells (MKN-28 and MKN-74) were higher than those of diffuse-type carcinoma cells (MKN-45 and KATO-III). To investigate whether the alpha 2 and alpha 6 integrin adhesion molecules are responsible for, or involved in carcinoma invasion. We further studied alpha 2 and alpha 6 expression patterns in these two types of cell line. Although fluorescence-activated cell sorting analysis revealed that all cells examined invariably expressed these integrin molecules, their expressional patterns were different among different cell lines. The intestinal-type carcinoma cells expressed integrins mainly along the cell-cell contact region, whereas the diffuse-type carcinoma cells showed a diffuse cytoplasmic pattern of integrin expression. Invasion by MKN-28, MKN-74 and MKN-45 cells through reconstituted basement membrane or type I collagen gel was significantly inhibited (P < 0.05) by 50 micrograms/ml anti-(alpha 2 integrin) or anti-(alpha 6 integrin) monoclonal antibodies. Our results suggest that active invasiveness is stronger in the intestinal-type than in the diffuse-type carcinoma cells and that alpha 2 and alpha 6 integrins play important roles in invasion of both types of gastric carcinoma cell lines. PMID- 9222297 TI - The in vitro effects of interleukin-12 upon tumor-infiltrating lymphocytes derived from renal cell carcinoma. AB - Clinical trials utilising interleukin (IL)-2 with tumor-infiltrating lymphocytes (TIL) have demonstrated efficacy in the treatment of metastatic renal cell carcinoma (RCC). Several cytokines, as well as growth factors have demonstrated modulatory effects upon the biological properties of TIL from RCC, suggesting a potentially important role for cytokines other than IL-2 in the development of active and tumor-specific TIL. IL-12 was recently characterized as a natural killer-cell-stimulatory factor or cytotoxic-T-cell-maturation factor. These properties of IL-12 prompted us to investigate the impact of this cytokine upon the activation of TIL from human RCC. In an attempt to enhance the in vitro growth and activity of renal TIL, we have grown eight renal TIL cultures in varying concentrations of IL-2 (8, 40, 80, 400 U/ml) and IL-12 (200 U/ml). In addition, IL-12 (200 U/ml) was added to TIL cultures pre-activated with IL-2 (400 U/ml). Growth, cell expansion, and the ability of TIL to release certain cytokines upon tumor stimulation were determined. Proliferation assays, phenotypic analysis, and cytotoxicity assays were performed at an early and a late culture stage. IL-12, alone and when added to suboptimal concentrations of IL-2, failed to induce TIL growth. While the addition of IL-12 to optimal doses of IL-2 suppressed TIL culture expansion, sequential culture exposure first to IL 2 and then to IL-2+IL-12 increased the number of cells expressing CD3+/CD56+ and these cultures demonstrated enhanced in vitro lysis of autologous tumor. IL-12 clearly demonstrated a sequence-dependent impact of the biological behaviour of TIL from RCC. The optimal use of IL-12 in the in vitro expansion of renal TIL may result in cells with an enhanced specific anti-tumor effect. PMID- 9222298 TI - Reconstruction and expression of chimeric anti-HBx antibody in Escherichia coli. AB - The variable regions of murine monoclonal anti-HBx immunoglobulin and the constant region of human antibody were cloned by reverse transcript-polymerase chain reaction (RT-PCR). The heavy-chain and light-chain variable regions were connected and coexpressed with human constant region C-r3 and C-k3 in the reconstructed vector of E. coli. The products showed high specificity and binding ability with HBx. Which is closely associated with hepatocarcinogenesis. This makes it possible to humanize the mouse monoclonal antibodies and express the fusion protein in E.coli for potential radioimmunotherapy in patients with hepatocellular carcinoma. PMID- 9222299 TI - Transformation-related expression of a low-molecular-mass tropomyosin isoform TM5/TM30nm in transformed rat fibroblastic cell lines. AB - We cloned a full-length rat TM5/TM30nm cDNA. Using this cDNA as a probe, we demonstrated that expression of TM5/TM30nm mRNA was higher in the tumorigenic rat fibroblastic cell lines SR-3Y1-2 and fos-SR-3Y1-202 than in the normal cell line 3Y1. High expression of TM5/TM30nm protein in SR-3Y1-2 and fos-SR-3Y1-202 cells was also detected by Western blot analysis using anti-TM5/TM30nm antiserum. In addition, the cellular localization of this protein differed between 3Y1 cells and tumorigenic ones. These findings suggest that TM5/TM30nm is involved in malignant transformation of rat fibroblastic cells. PMID- 9222300 TI - A rapid, useful and quantitative method to measure telomerase activity by hybridization protection assay connected with a telomeric repeat amplification protocol. AB - Telomerase, a ribonucleoprotein enzyme, is expected to be a new marker for cancer diagnosis. TRAP (the telomeric-repeat amplification protocol) developed by Kim et al. is a sensitive method to detect telomerase activity. Telomerase activity is detected by TRAP in most malignant cells in vivo and in vitro, but it is not found, or found only in very low amounts, in normal somatic cells and tissues. TRAP and its modified protocols are, however, not always suitable for measuring the activity of a large number of clinical samples to diagnose cancer, because they generally require a time-consuming detection step such as gel electrophoresis with radioactive materials. To improve the procedure for mass diagnosis, we applied a hybridization protection assay (HPA) to replace the detection step. HPA, which employs an acridinium-ester-labelled probe, is radioactivity-free, easy to handle without electrophoresis, quick, and applicable to a quantitative format. In this work we have established and demonstrated the advantages of TRAP/HPA. The telomerase activity of various primary and established cells, differentiating cancer cells, and normal and tumour colorectal and liver tissues was quantitatively analysed by TRAP/HPA. The results indicate that HPA combined with TRAP is a rapid and simple method, easy to handle and quantify, for the clinical diagnosis of cancer. PMID- 9222301 TI - Enhancement of tumor cell susceptibility to tumor-infiltrating lymphocytes by cisplatin. AB - Some means of enhancing the susceptibility of tumor cells to tumor-infiltrating lymphocytes (TIL) are required in adoptive immunotherapy. This study was designed to investigate whether or not tumor cell lysis by TIL was enhanced by treatment of the tumor cells with cisplatin, and also to clarify the mechanism of cisplatin's action on tumor cells. Autologous tumor cells and established cancer cell lines, including KATO-III and MKN-28, were used. Cytotoxic activities of TIL, the surface antigens of tumor cells, conjugation of TIL and tumor cells, and the production of TNF alpha from TIL were analyzed. Tumor cells treated with 2 micrograms/ml cisplatin for 12 h in vitro were more susceptible to bulk-cultured TIL and TIL clones. The surface antigens of tumor cells were not altered by the treatment with cisplatin. Cisplatin-treated tumor cells showed a higher binding ratio to TIL than did non-treated tumor cells. The anti-(tumor necrosis factor) (anti-TNF) or anti-TNF receptor antibody blocked the enhancement of cytotoxic activity by cisplatin. Thus, it was clarified that cisplatin enhanced the susceptibility of tumor cells to bulk-cultured TIL and TIL clones. Furthermore, the enhancement of cytotoxic activity by TIL in cisplatin-treated tumor cells was caused by a higher binding ratio to TIL and higher susceptibility to the TNF produced by TIL. PMID- 9222302 TI - P-glycoprotein expression in soft-tissue sarcomas. AB - Multidrug resistance (MDR) is an important problem in chemotherapy for neoplastic disease. In humans. MDR is mainly mediated by P-glycoprotein (P-gp) a product of the MDRI gene, which acts as a transmembrane protein pump and eliminates chemotherapeutic agents from the cells. Expression of P-gp was immunohistochemically studied by using two monoclonal antibodies, JSB-1 and C 219, on paraffin-embedded sections from 55 patients with soft-tissue sarcoma. The histological diagnosis of tumors was malignant fibrous histiocytoma in 24 cases, liposarcoma in 9, synovial sarcoma in 7, malignant neurogenic tumors in 6, leiomyosarcoma in 5, others in 4. The histological grade was determined on the basis of criteria previously proposed by us. Out of 55 cases, 34 (62%) were positive for P-gp expression. There was a significant difference in P-gp expression between high-grade (90%) and intermediate and low-grade tumors (46%) (P < 0.005). Tumors expressing P-gp had a less favorable prognosis than P-gp negative tumors in the high- and intermediate-grade tumors. The current study demonstrated that the estimation of P-gp expression could be used to select appropriate therapeutic modalities. PMID- 9222303 TI - Prospective study of early detection for primary liver cancer. AB - PURPOSE: To determine whether repeated screening can lead to early detection of primary liver cancer (PLC) and in turn to an improved clinical result. METHODS: In this randomized controlled study, Shanghai urban residents aged 35-55 years and with serum evidence of HBV infection or chronic liver disease were eligible for recruitment. Using cluster sampling, these subjects were allocated into two groups-the screening group and the control group: there were 8109 subjects in the screening group and 9711 in the control group. Subjects in the screening group were tested with serum AFP and real-time ultrasound every 6 months. One to four rounds of screening were completed. Liver cancer was treated according to stage at diagnosis. RESULTS: All subjects enrolled were followed up and classed at the end-point as alive without liver cancer, alive with liver cancer, dead from liver cancer, or dead from another cause. The mean follow-up was 1.2 years; total follow-up was 12,038 person-years in the screening group and 9,573 person-years in the control group. We detected 38 patients with PLC in the screening group and 18 patients with PLC in the control group. In the patients in the screening group 76.8% of patients were at a subclinical stage, and 70.6% of them underwent resection, the 1- and 2-year survival rates being 88.1% and 77.5%, respectively. However, in the control group, none of the patients was at a subclinical stage when diagnosed, none of them underwent resection, and none of them survived over 1 year. The lead time was estimated at 0.45 years. The cost of detecting PLC at an early stage was RMB 12,600 (US$1,500). CONCLUSION: The study proved that screening the high-risk population for PLC with a serum AFP test and real-time ultrasound examination can detect patients in the early stages, increase the resection rate and prolong the survival time. It is therefore recommended that screening for PLC be advocated in any high-risk area. PMID- 9222304 TI - Effects of 1,25-dihydroxyvitamin D3 on tumor cell invasion to the extracellular matrix in human fibrosarcoma HT1080 cells and its correlation with laminin. AB - We investigated the role of the active form of vitamin D, 1,25-dihydroxyvitamin D3[1,25(OH)2D3], in promoting tumor cell invasiveness through the extracellular matrix, and showed that 1,25(OH)2D3-induced reduction of laminin production by the cells was correlated with the inhibitory effect of the hormone on tumor cell invasiveness. 1,25(OH)2D3 significantly inhibited invasiveness through the matrix, type IV collagenolytic and migratory activity, but not cell attachment to the matrix in human fibrosarcoma HT1080 cells. The 1,25(OH)2D3-induced inhibition showed the same dose dependency and magnitude for invasiveness as for the effects on type IV collagenolysis and cell migration. 1,25(OH)2D3 inhibited laminin production from the cells in a dose-dependent manner. The inhibitory effects of 1,25(OH)2D3 on the invasiveness, type IV collagenolysis and cell migration appeared to parallel the hormone-induced reduction of laminin production. Antilaminin monoclonal antibody, blocking the activity of laminin in the culture medium, inhibited HT1080 cell invasiveness. In the presence of exogenous laminin, 1,25(OH)2D3-induced inhibition of invasion was not observed. These findings suggest that 1,25(OH)2D3 acts on HT1080 cells, inhibiting the expression of laminin from the cells, and that the reduced laminin expression leads to the inhibition in the type IV collagenolytic and migratory activity of the cells, and consequently, to the inhibition of invasiveness through the extracellular matrix. PMID- 9222305 TI - Is free prostate-specific antigen helpful in the differential diagnosis of benign hyperplasia and cancer of the prostate? AB - PURPOSE: We studied the clinical relevance of the determination of free prostate specific antigen (f-PSA) in addition to total PSA (t-PSA). METHODS: Both t-PSA and f-PSA values of frozen sera obtained pretherapeutically from 80 patients with prostate carcinoma (PC) and 171 patients with benign hyperplasia of the prostate (BPH) were analyzed by means of the Tandem-E PSA and Tandem-R f-PSA immunoassays (Hybritech, San Diego, Calif.). RESULTS: At 95% specificity, a cutoff value of 15.7 micrograms/l was obtained for t-PSA [9 patients with BPH (5%) were above this value]. For this cutoff value, we calculated a sensitivity of 50% (40 patients with PC were above this value). Using the same criteria for the ratio (Q) f-PSA:t-PSA a cutoff of 0.086 was found again at a specificity of 95%. In a second step, only patients with t-PSA values below the cutoff level of 15.7 micrograms/l were considered. Out of these patients, 6 of 156 with BPH (missing values = 6) and 11 of 40 with PC were below the above-mentioned ratio (Q = 0.086). Therefore, sensitivity was 28% for this subgroup. Considering both steps (t-PSA and Q) 51 patients with PC were detected correctly and 15 patients with BPH would have undergone biopsy unnecessarily (positive biopsy rate: 77%). CONCLUSIONS: High t-PSA levels are very good indicators for the presence of PC. There is still concern for improving the differentiation between BPH and PC, when an intermediate or low value (< or = 95% specificity) is observed. The determination of Q is only useful in this range and is helpful for the clinician's decision whether to apply or avoid biopsy. PMID- 9222306 TI - Serum levels of intercellular adhesion molecule-1 in squamous cell carcinoma of the head and neck. AB - Soluble intercellular adhesion molecule-1 (s-ICAM-1) was measured in the sera of 131 patients with primary and 50 patients with recurrent squamous cell cancer of the head and neck (HNSCC). 30 patients with benign ear, nose and throat diseases served as controls. s-ICAM-1 levels in serum are high in patients with HNSCC, particularly in the advanced tumor stages (UICC IV). Highest levels can be measured at the time of tumor recurrence and locoregional lymph node metastases. The sensitivity (95% specificity) of s-ICAM-1 (cutoff-level: 473 ng/ml) is 4% at primary diagnosis and 12% for recurrent disease. A coefficient of correlation for s-ICAM-1 in combination with SCC, carcinoembryonic antigen and CYFRA 21-1 indicates that no correlation can be found of s-ICAM-1 compared with traditional tumor markers. Due to overlapping values in control and patient groups s-ICAM-1 is not suitable for a specific clinical use in HNSCC. PMID- 9222307 TI - Infrequent alterations of the p16 (MTS-1) gene in human gastric cancer. AB - p16 (MTS-1, multiple tumor suppressor gene 1), a putative tumor suppressor gene, is one of the cyclin-dependent kinase inhibitors (CDI) and it regulates the G1/S transition of the cell cycle. To clarify the role of p16 in primary gastric cancer, we have investigated somatic mutations of this gene by using the polymerase chain reaction/single strand conformation polymorphism (PCR-SSCP) method. In 23 surgical specimens of primary gastric cancer, none were detected in exon1 and exon 2. Among the 6 human gastric cancer cell lines examined, PCR products were not found in 2, MKN28 and MKN45, suggesting the presence of homozygous deletions. No mutation was found in the other 4 cell lines. Furthermore, decreased expression levels were not observed in 13 gastric cancer tissues by reverse transcription PCR (RT-PCR). Considering the above results of PCR-SSCP and RT-PCR, genetic alterations of the p16 gene are rarely implicated in human gastric cancer tumorigenesis. PMID- 9222308 TI - Ascitic growth of a T cell lymphoma in mice alters the humoral and cellular immune response to exogenous antigens. AB - The effect of the ascitic growth of Dalton's lymphoma (DL), a T cell lymphoma, on the immune responses of the host mice to exogenous antigens, with respect to the humoral response, delayed-type hypersensitivity (DTH) response and the antigen presenting ability of macrophages was investigated. The humoral immune response to sheep red blood cells (SRBC) as well as the antigen-presenting ability of macrophages (with keyhole limpet hemocyanin as the standard antigen) in the DL bearing mice were consistently higher than in the normal mice, although the magnitude showed a decline during later tumor stages. However, the DTH response to SRBC was suppressed in the DL-bearing mice compared with the response in the normal mice. The possible mechanisms are discussed. In vivo administration of FK565, a synthetic biological response modifier, enhanced the humoral immune response as well as the antigen-presenting ability of the macrophages in the normal and early DL-bearing mice, whereas these immune responses n the later tumor-bearing animals were found to be nonresponsive to FK565 treatment. In contrast, the DTH response in the normal as well as in the DL-bearing mice was suppressed on FK565 administration. This is the first study of its kind regarding the effect of the ascitic growth of any T cell lymphoma on various aspects of the immune response to exogenous antigens and the correlation thereof with an immunomodulator. PMID- 9222309 TI - Characterization of a liver metastatic variant of murine colon 26 carcinoma cells. AB - Intraportal vein injection of highly metastatic L5 cells consistently resulted in liver metastases (increases in the number of tumor colonies in the liver), whereas inoculation of P cells rarely did. L5 cells invaded the basement membrane Matrigel in greater numbers than did P cells, suggesting that the metastatic potential of L5 cells is partly related to enhanced invasive properties. The enhanced adhesion of L5 cells to fibronectin-, laminin- and Matrigel-coated substrates, as well as their haptotactic migration to fribronectin, may be associated with the preferential expression of VLA-2 and VLA-4 integrins on the surface of these cells detected by flow cytometry. Gelatin zymograms showed that the degradative activity of 72-kD gelatinases was greater in L5 cells than P cells. These results indicate that, in addition to adhesiveness and motility, the invasive ability of L5 cells may also be attributed to enhanced gelatinolytic activity. L5 cells grew more rapidly than P cells in vitro. Thus, an experimental model using highly metastatic colon 26 L5 cells would be useful for analyzing the molecular mechanism of liver metastasis and for evaluating the efficacy of treatment of occult micrometastases which may already have been disseminated at the time of surgery. PMID- 9222310 TI - Prostate-specific antigen and early detection of prostate cancer. AB - Prostate-specific antigen (PSA) is biochemically a 33-kDa serine protease and is the prototype of tumor marker most useful in investigating basic science and clinical application of prostate cancer. As an immunohistopathological marker, PSA is especially effective in the identification of distant metastatic prostate carcinoma and in the differential diagnosis of poorly differentiated transitional cell carcinomas of the bladder from prostate carcinoma. As a serologic marker, PSA is most useful in staging, monitoring and in early detection of recurrent disease. The greatest value of PSA is as a screening aid along with digital rectal examination for early detection of prostate cancer. PSA-based screening tests have been performed in various regions of the world and have yielded prostate cancer detection rates proportional to that of incidence rates in the countries. Most significantly, PSA detects early nonpalpable prostate cancer. Approximately 85% of the prostate tumors detected through PSA have the clinical features associated with medically important cancer. Further, a majority of the tumors, 70%, are confined to the organ. Although the issues of cost-benefit and increase in ultimate survival rate are still being evaluated in two large-scale randomized national trials, timely and definitive therapies of these potentially lethal tumors can result in high cure rate and improve the quality of patients' life. PMID- 9222311 TI - Isolation of the Rieske iron-sulfur protein from the cytochrome b6/f complex of spinach chloroplasts. PMID- 9222312 TI - Spirodela oligorhiza chloroplast DNA codes for ATPase subunits alpha and beta. Immunological evidence from a coupled transcription-translation system. PMID- 9222313 TI - Modification of 18 S rRNA in the 40 S ribosomal subunit of yeast with dimethyl sulfate. PMID- 9222314 TI - Analysis of Euglena gracilis chloroplast DNA. The DNA fragment EcoRI . N carries genetic information for a 53,000 M(r) polypeptide. PMID- 9222316 TI - The compaction of mouse heterochromatin as studied by nuclease digestion. PMID- 9222317 TI - A pitfall in the interpretation of data on adenylate cyclase inactivation by irradiation. PMID- 9222315 TI - A rapid purification by affinity chromatography of orotate phosphoribosyltransferase from Escherichia coli K-12. PMID- 9222318 TI - Conformational flexibility in enkephalins: solvent dependent transitions in peptides with Gly-Gly segments detected by circular dichroism. PMID- 9222319 TI - Incorporation of melittin into phosphatidylcholine bilayers. Study of binding and conformational changes. PMID- 9222321 TI - Thyrotropin releasing hormone stimulates metabolism of phosphatidyl inositol in GH3 cells. A possible mechanism in stimulus-response coupling. PMID- 9222320 TI - Undersulfated beta-D-xyloside glycosaminoglycans are secreted in the presence of the ionophore monensin. PMID- 9222322 TI - Modification of amino groups in Candida utilis uricase with naphthoquinone disulfonic acid in relation to the enzymic activity. PMID- 9222323 TI - New substrates of acetylcholinesterase. PMID- 9222324 TI - Sequence of Escherichia coli D-serine dehydratase. Location of the pyridoxal phosphate binding site. PMID- 9222325 TI - Existence of two different species of mitochondrially translated proteolipids in ATPase complex of yeast mitochondria. PMID- 9222326 TI - Further characterization of the discontinuities in cauliflower mosaic virus DNA. PMID- 9222327 TI - Differences in polyamine metabolism of the undifferentiated and differentiated neuroblastoma cells. Metabolic labeling of an 18,000-M(r) protein by [14C]putrescine and the conversion of putrescineto GABA. PMID- 9222328 TI - Myoblast aminophospholipid asymmetry differs from that of fibroblasts. PMID- 9222329 TI - Evidence for an essential role for high-density lipoprotein in progesterone synthesis by rat corpus luteum. PMID- 9222330 TI - Receptor-mediated endocytosis of alpha 2macroglobulin-protease complexes by fibroblasts in culture. Competitive inhibition by bacitracin. PMID- 9222331 TI - Strontium ions stimulate phosphoinositide metabolism in human blood platelets. PMID- 9222333 TI - Evidence that a specific succinic semialdehyde reductase is responsible for gamma hydroxybutyrate synthesis in brain tissue slices. PMID- 9222334 TI - [Rapid induction of hepatic acetyl-CoA carboxylase activity after estradiol benzoate injection in the quail]. AB - The time course of hepatic Acetyl-CoA carboxylase activity as well as hepatic and plasmatic fatty acids concentrations following a single injection of estradiol benzoate (EB, 0.2 mg/kg) was studied in the quail. Acetyl-CoA carboxylase activity increases rapidly and reaches its peak 3 h after the injection of EB. Similarly, hepatic and plasmatic fatty acids concentrations are significantly increased 6 h after the hormonal injection and attain their highest level 18 h later. These results suggest that estrogen affects the hepatic fatty acids biosynthesis by regulating the conversion of acetyl-CoA to malonyl-CoA. PMID- 9222332 TI - The metabolic production of 4-methylumbelliferone and its beta-D-glucuronide from the corresponding beta-D-xyloside by fibroblasts in culture. PMID- 9222335 TI - Phosphorylated sites of chicken erythrocyte histone H5 by a cyclic AMP independent protein kinase from mouse plasmocytoma cells. PMID- 9222337 TI - Neurovirology: evolution of a new discipline. PMID- 9222336 TI - Study of in vitro phosphorylation of histones H3, H4 and of the non-acetylated and acetylated tetramers (H3-H4)2. PMID- 9222338 TI - Progressive multifocal leukoencephalopathy: the evolution of a disease once considered rare. AB - Progressive multifocal leukoencephalopathy, a formerly rare disease that chiefly occurred in persons with underlying lymphoma and chronic lymphocytic leukemia, is now seen with increasing frequency in the era of acquired immunodeficiency syndrome. Progressive multifocal leukoencephalopathy is currently estimated to arise in 5% of all human immunodeficiency virus-infected individuals. The clinical features of the disorder in patients with acquired immunodeficiency syndrome do not appear to be significantly different from progressive multifocal leukoencephalopathy occurring in association with other immunosuppressive disorders. Radiographically, the appearance of HIV dementia on magnetic resonance imaging is sometimes confused with that of progressive multifocal leukoencephalopathy. Among the characteristics that are helpful in distinguishing between the two disorders are the presence of focal findings, the rate of disease progression, the specific magnetic resonance imaging attributes, including the location of the lesions, and certain cerebrospinal fluid parameters, including surrogate markers for human immunodeficiency virus dementia and the presence of myelin basic protein. The remarkable increase in the burden of progressive multifocal leukoencephalopathy has provided a vital impetus for its study, particularly with respect to diagnosis and therapy. Establishing an unequivocal diagnosis of progressive multifocal leukoencephalopathy currently requires brain biopsy. The application of polymerase chain reaction for JC virus amplification to cerebrospinal fluid samples suggests that it may provide an alternative means of diagnosis. Recent in vitro studies of cytosine arabinoside and camptothecin suggest that they, or similar agents, may prove useful in the treatment of this illness and well-designed clinical trials are underway. PMID- 9222339 TI - Herpes simplex virus latent infection in the nervous system. AB - Herpes simplex virus (HSV) establishes a latent infection in the human peripheral nervous system and can cause recurrent disease by reactivation. Intensive effort has been directed in recent years to unveil the molecular, cellular and immune mechanisms, as well as the virus-host interactions associated with latent HSV infection. The aim of this review is to summarize current knowledge regarding the site of latent infection, the molecular phenomena of latency, and the mechanisms of the various stages of HSV-1 latent infection in the nervous system, relating them where possible to the human situation. Specifically, the following biological questions are addressed: (1) How does this lytic virus survive in the nervous system and why can it establish a lifelong latent infection in nerve cells? (2) What advantage is conferred on HSV by establishing latent infection in nervous tissue? (3) What can be gathered from the accumulated knowledge on latency about the pathogenesis of herpes simplex encephalitis? PMID- 9222340 TI - Cellular localization of human herpesvirus-6 in the brains of children with AIDS encephalopathy. AB - Human herpesvirus-6, the etiologic agent of exanthem subitum, is a ubiquitous virus that infects almost all children by the age of 2 years and that has previously been shown to be neuroinvasive. These characteristics suggest that human herpesvirus-6 may be important in the neuropathogenesis of acquired immune deficiency syndrome (AIDS) in children. To address this hypothesis, we evaluated postmortem pediatric brain tissues for the presence of human herpesvirus-6 infection. Using in situ hybridization with a digoxigenin-labeled DNA probe for the large tegument protein gene of human herpesvirus-6, we detected nuclear signals in postmortem brain tissue from 4/5 children with human immunodeficiency virus-1 encephalitis. Human herpesvirus-6 DNA was found in numerous oligodendrocytes of the white matter and less frequently in astrocytes, macrophages, microglia and neurons. The human herpesvirus-6 positive cells detected by in situ hybridization were not immunoreactive either for human herpesvirus-6 early nuclear phosphoproteins or for surface glycoproteins associated with productive infection. Only rare human herpesvirus-6 infected cells were found in age-matched control brain tissues. No human herpesvirus-6 infected cells were found in human fetal brain tissue. These data suggest that human herpesvirus-6 is more extensively disseminated in neural cells in the presence of human immunodeficiency infection and immunodeficiency in pediatric AIDS patients, and it may contribute to the pathogenesis of AIDS encephalopathy. PMID- 9222341 TI - Evaluation of the role of cytokine activation in the multiplication of JC virus (JCV) in human fetal glial cells. AB - The human polyomavirus, JCV, is the etiologic agent of the fatal central nervous system demyelinating disease, progressive multifocal leukoencephalopathy. Progressive multifocal leukoencephalopathy occurs most frequently in patients with underlying immunosuppressive disorders and is the direct result of virus multiplication in oligodendrocytes, the myelin producing cell in the central nervous system. In this report we test the ability of cellular activation signals to modulate expression of the JCV genome in either transfected or infected human fetal glial cells. In addition, we analyze the binding of nuclear proteins isolated from untreated and cytokine treated human fetal glial cells to transcription factor binding sites in the JCV regulatory region. In contrast to the effects of cellular activation on the expression of the HIV-1 promoter in these cells, none of the cellular activators tested increased expression of JCV. The cytokine, TNF-alpha, increased binding of NF kappa B (p50/p65) to a JC NF kappa B site but did not modulate the binding of nuclear proteins to the overlapping NF-1/AP1 region of the JCV enhancer. When taken together these results suggest that the response of JCV to cellular activation signals may be fundamentally different from the response of HIV-1 to these signals in human fetal glial cells and that the JC NF kappa B site may not be required for JCV gene expression or multiplication in vivo. PMID- 9222342 TI - HTLV-I-associated myelopathy: analysis of 213 patients based on clinical features and laboratory findings. AB - We studied the clinical features and laboratory findings in 213 patients with HTLV-I-associated myelopathy/tropical spastic paraparesis as diagnosed in Kagoshima University Hospital. Some aspects of clinical features in HTLV-I associated myelopathy/tropical spastic paraparesis were characterized by mode of HTLV-I transmission and age of onset. The patients with onset after 15 years old and no history of blood transfusion before the onset of the disease (151 patients, group I) showed a shorter interval between the time of disease onset and that of inability to walk. The patients with onset before 15 years old and without history of blood transfusion (21 patients, group II) had short stature and slow progression of the disease. The interval time and the progression of the disease in patients with history of blood transfusion before onset of disease (41 patients, group III) were in between those of the above two groups. Patients whose ages of onset were older than 61 years old showed a faster progression than those with younger onset regardless of the mode of HTLV-I transmission. HTLV-I associated myelopathy/tropical spastic paraparesis patients often also showed other organ disorders such as leukoencephalopathy (69%), abnormal findings on chest X-ray (50%), Sjogren syndrome (25%) and arthropathy (17%). The patients with low anti-HTLV-I antibody titers in the cerebrospinal fluid (2X-8X by PA method) had an older age of onset on average, milder clinical symptoms and lesser increase of neopterin in the cerebrospinal fluid than those in the high titer subgroup whose titers were higher than 1024X in cerebrospinal fluid regardless of the mode of HTLV-I transmission. We speculate that the clinical course of HTLV-I associated myelopathy/tropical spastic paraparesis mainly shows a slow progression which consists of an initial progressive phase (probably an inflammatory phase) and a latter chronic phase, although some patients showed acute/subacute onset and rapid progression. PMID- 9222343 TI - Characterization of a glial cell-specific DNA-protein complex formed with the human T cell lymphotropic virus type I (HTLV-I) enhancer. AB - Human T cell lymphotropic virus type I (HTLV-I) encodes the trans-activator, Tax, which facilitates viral transcription from three 21 bp repeated elements within the U3 region of the long terminal repeat (LTR). Electrophoretic mobility shift (EMS) analyses utilizing double-stranded (ds) oligonucleotides homologous to each of the 21 bp repeats and nuclear extracts derived from selected cell lines of lymphocytic, neuronal, and glial origin have demonstrated the differential binding of cellular factors to each of the three 21 bp repeats. Specifically, both a glial cell-specific DNA-protein complex (designated GCS) and 21 bp repeat specific DNA-protein complexes (designated U1 and U2) were detected. The formation of the GCS DNA-protein complex may involve activating transcription factor (ATF)/cAMP-response element (CRE) binding protein (CREB) family member(s) while the formation of the U1 and U2 DNA-protein complexes may involve an Sp1 related factor. In addition, three ATF-CREB-related DNA-protein complexes common to each individual 21 bp repeat (designated C1-C3) were also detected. However, we demonstrated that the abundance of the C1 and C2 DNA-protein complexes detected with the individual 21 bp repeats and glial cell nuclear extract was relatively low compared to that obtained with lymphocyte, monocyte, or neuronal nuclear extracts. We also have demonstrated that the ATF-CREB factors participating formation of the GCS DNA-protein complex are distinct from those participating in formation of the C1-C3 DNA-protein complexes. Based on nucleotide sequence requirements and immunoreactivity, we suggest that the GCS DNA-protein complex may contain a novel glial cell type specific ATF-CREB-related factor(s). Furthermore, we demonstrate that the CRE modulator (CREM) protein in conjunction with the ATF/CREB factor, CREBP1, interact with each of the three 21 bp repeats to form the C3 DNA-protein complex. However, the abundance of the C3 DNA-protein complex formed utilizing the promoter proximal repeat is dramatically lower compared to either of the other two 21 bp repeat elements. Based on these observations, we suggest that the differential binding of cellular factors to each of the three 21 bp repeat elements may play a role in basal as well as Tax mediated LTR-directed transcription within cell populations of either immune or nervous system origin. PMID- 9222344 TI - Pathogenesis of lymphocyte-tropic and macrophage-tropic SIVmac infection in the brain. AB - SIVmac239 replicates productivity in activated CD4+ T lymphocytes, but inefficiently in macrophages from rhesus macrophages. Inoculation of the virus into animals results in an acute, highly productive burst of virus replication in activated T lymphocytes in lymphoid tissues and infected cells invade the central nervous system (CNS). This phase lasts a few weeks and is eventually followed by development of immunosuppression of different degrees of severity, opportunistic infections, and tumors related to the loss of T lymphocytes. On rare occasions, infected immunosuppressed animals develop encephalitis and/or interstitial pneumonia, syndromes that are associated with selection of mutant viruses that replicate efficiently in macrophages of these tissues. Usually, however, brains of animals dying with AIDS caused by SIVmac239 appear histologically normal. Is the brain infected with virus? We report here on a macaque dying with AIDS, a neuroinvasive tumor and interstitial pneumonia associated with macrophage-tropic virus. Except for focal infiltration of tumor cells, the brain was normal histologically. We examined the virus and viral DNA from different tissues and found that lymphocytes but not macrophages from lymph nodes and spleen yielded virus, whereas macrophages but not lymphocytes from the lung produced virus. No virus was recovered from the brain but small amounts of viral p27 were present in the brain homogenate. Viral sequences were present in the brain as determined by PCR from tissue DNA. Comparison showed that the viral sequences in the brain closely resembled those from the spleen. Presumably, the virus caused a minimally productive infection detectable by production of small amounts of p27, but was not accompanied by any histopathological changes. It is unclear why the macrophage-tropic virus in the lung failed to 'take-off' in the brain of this animal. To determine whether this virus had encephalitic potential, we inoculated the lung homogenate containing cell-free, macrophage tropic virus into a young pigtail macaque, a species known to be sensitive to primate lentiviral infections. This animal developed severe encephalitis 10 weeks later. Virus from the brain was very similar to the inoculum virus, proving its encephalitic potential. Possible reasons for the differences in neurovirulence of this virus between the two animals remain speculative. PMID- 9222345 TI - Measles virus-specific dsRNAs are targets for unwinding/modifying activity in neural cells in vitro. AB - Biased hypermutation events found predominantly in the matrix gene of measles virus isolated from persistent human CNS infections have been attributed to the action of a cellular unwinding/modifying activity (UMA). To define the level and distribution of this activity in brain cells, fractionated extracts were prepared from the nuclei and cytoplasm of human glioblastoma (D-54, U-251) and neuroblastoma (IMR-32, SKN-MC) cells and analyzed for their ability to modify synthetic dsRNAs specific for the measles virus (MV) matrix (M) gene. On a quantitative basis we could show that the activity localized to both the nuclear and cytoplasmic compartments of both cell types analyzed independent of cell proliferation. The presence of significant levels of UMA in the cytoplasm of human brain cells following growth arrestment in vitro with retinoic acid supports the interpretation that UMA may contribute to the attenuation of MV gene functions during the primary infection of brain cells, thereby supporting the establishment of virus persistence. PMID- 9222346 TI - The effect of Theiler's murine encephalomyelitis virus (TMEV) VP1 carboxyl region on the virus-induced central nervous system disease. AB - Members of the Theiler's murine encephalomyelitis virus GDVII subgroup, which includes GDVII strain, are highly neurovirulent and induce a rapidly fatal polioencephalomyelitis. By contrast, Theiler's original subgroup members, which includes DA strain, are not as neurovirulent, and produce a chronic, demyelinating disease with virus persistence. We investigated the importance of the carboxyl region of the capsid protein VP1 in TMEV-induced disease since a trypsin-cleavable immunodominant neutralization epitope is situated in the VP1 carboxyl region, and since this region is believed to lie adjacent to the putative receptor binding site. The present studies support the role of DA VP1 residue 268 (and the aligned GDVII VP1 270) in Theiler's murine encephalomyelitis virus-induced CNS disease; however, the effect of this residue varies depending on its context: mutation of DA VP1 268 attenuates demyelination; mutation of GDVII VP1 270 in a GDVII/DA recombinant virus has no effect on demyelination but reduces early deaths (neurovirulence); mutation of GDVII VP1 270 in GDVII virus has no effect on neurovirulence. These data suggest that DA VP1 268/GDVII VP1 270 are not functionally equivalent and that a residue in recombinant viruses can differ in function from the same residue situated in a parental strain. Additional mutagenesis studies suggest that: the trypsin cleavage site of TMEV, which affects virus viability, is located at the lysine at DA VP1 261 (GDVII VP1 263); GDVII VP1 276, the predicted carboxyl terminus of VP1, affects VP1/2A processing and virus infectivity. PMID- 9222348 TI - Effect of neurotropic virus infection on neuronal and inducible nitric oxide synthase activity in rat brain. AB - To elucidate the potential role of inducible nitric oxide synthase (iNOS) and neuronal constitutive nitric oxide synthase (cNOS) in the pathogenesis of virus induced encephalopathy, the activities of both NOS isoforms were determined in the brains of rats infected with Borna disease virus (BDV) or rabies virus. iNOS activity strongly increased, whereas neuronal cNOS activity significantly decreased in a time-dependent manner after either BDV or rabies virus infection. Choline acetyltransferase activity in the brain remained unchanged during both virus infections, suggesting that the decrease in cNOS activity does not reflect a generalized neuronal loss. Immunohistochemistry and Northern blot analyses indicate that the decrease in neuronal cNOS activity is due to a decrease in cNOS protein and mRNA synthesis. These results suggest that both an excessive generation of NO by activated macrophages or microglia, as well as a decrease of NO production in neurons may contribute to the neuropathogenesis of neurotropic virus infections. PMID- 9222347 TI - H-2 Dd transgene suppresses Theiler's virus-induced demyelination in susceptible strains of mice. AB - Theiler's murine encephalomyelitis virus is a picornavirus which induces chronic immune-mediated central nervous system demyelination and virus persistence in susceptible strains of mice. Using murine strains with congeneic recombinant haplotypes, the H-2D region within the class I major histocompatibility complex has been shown to be important in determining susceptibility/resistance to chronic Theiler's murine encephalomyelitis virus infection. We examined the role of H-2D in demyelinating disease with the use of transgenic D8 mice (H-2Dd, resistant haplotype) crossed to susceptible B10.Q (H-2q) and B10.S (H-2s) mice. Expression of the H-2Dd transgene dramatically suppressed demyelination and reduced the number of virus-antigen positive cells in the spinal cord 45 days following infection. More complete protection was observed in transgenic B10.Q (D8+) mice than in transgenic B10.S (D8+) mice. These experiments support the hypothesis that the immunologic basis of resistance by H-2D is determined by effective antigen presentation which prevents virus persistence and subsequent demyelination. PMID- 9222349 TI - Assessment of neuronal density in the putamen in human immunodeficiency virus (HIV) infection. Application of stereology and spatial analysis of quadrats. AB - Human immunodeficiency virus causes neuronal loss in various brain regions, but it has not been reported in the putamen. However, decrease in the volume of the putamen has been observed by magnetic resonance imaging. In order to clarify this issue two complementary methods; the stereological probe, the disector, and spatial analysis of quadrats, were applied in nondemented individuals who had died of acquired immune deficiency syndrome. A 21% decrease in neuronal density was observed in the human immunodeficiency virus group, especially those cases with human immunodeficiency virus encephalitis; however the statistical significance of this finding was borderline. PMID- 9222350 TI - Persistence of varicella-zoster virus DNA in elderly patients with postherpetic neuralgia. AB - The most common complication of zoster in the elderly is postherpetic neuralgia, operationally defined as pain persisting longer than 1-2 months after rash. The cause of postherpetic neuralgia is unknown. Using polymerase chain reaction, we detected varicella zoster virus DNA in blood mononuclear cells from 11 of 51 postherpetic neuralgia patients, but not in any of 19 zoster patients without postherpetic neuralgia, or in any of 11 elderly individuals without a history of zoster. Blood mononuclear cells from nine of 27 serially-bled postherpetic neuralgia patients were positive for varicella zoster virus DNA; six were positive once, and three patients were positive more than once. Our results indicated that postherpetic neuralgia may be related to persistence of varicella zoster virus. PMID- 9222351 TI - Herpes simplex virus replication hits a nerve. PMID- 9222353 TI - Induction of HLA class II in HTLV-I infected neuronal cell lines. AB - Human T-lymphotropic virus-I (HTLV-I) has been etiologically linked with HTLV-I associated myelopathy/tropical spastic paraparesis (HAM/TSP), a neurologic disease. The characteristic pathological finding in HAM/TSP is marked mononuclear infiltration of the CNS with destruction of the long tracts of the spinal cord. An increased expression of HLA surface antigens and cytokines in the CNS is associated with this inflammatory response. Furthermore, there is evidence for the presence of HTLV-I in HAM/TSP CNS specimens using in situ hybridization and polymerase chain reaction techniques. The relationship between HTLV-I infection of CNS cells and the observed upregulation of surface antigens in the CNS is not well understood. It has been previously demonstrated that HTLV-I infection of neuroblastoma cells leads to induction of HLA surface antigens. As an extension of these studies, HFGC and HCN-1a, neuronal cell lines of nontumorigenic origin, were infected with HTLV-I and the effect on HLA upregulation was studied. Infection of the neuronal cells was demonstrated by the presence of HTLV-I gp46 surface antigen on CD4 negative cells and by the in situ presence of HTLV-I RNA in neurofilament positive cells. Concurrent to HTLV-I infection, HLA class II surface antigen was observed on neurofilament positive cells. Upregulation of HLA class II was not observed in neuronal cells grown in the presence of interferon gamma or tissue necrosis factor-alpha. PMID- 9222352 TI - Mutation of a major histocompatibility class I locus, H-2D, leads to an increased virus burden and disease susceptibility in Theiler's virus-induced demyelinating disease. AB - Genetic studies have demonstrated that susceptibility to Theiler's murine encephalomyelitis virus (TMEV)-induced demyelinating disease is multigenic with linkage to the MHC class I locus, H-2D. We have analyzed the effect of mutations (H-2bm13 and H-2bm14) in the H-2Db gene on central nervous system (CNS) virus replication, virus-specific delayed type hypersensitivity (DTH) and disease induction in mutant [bm14D2F1 and bm13D2F1] and parental B6D2F1 hybrids. The results indicate that substitutions of only a single residue (bm14D2F1) or three residues (bm13D2F1) in H-2D in the mutant leads to a sequence of events culminating in disease susceptibility. Mutation of the H-2D gene is associated with reduced or delayed virus clearance following the acute phase of exponential CNS virus growth and an increased level of virus persistence. Concomittant with the greater virus antigen burden, mutant mice respond with higher levels of virus specific DTH and develop inflammatory demyelinating lesions. PMID- 9222354 TI - Neuronal sprouting in mouse sensory ganglia infected with herpes simplex virus type 2 (HSV-2): induction of growth-associated protein (GAP-43) and ultrastructural evidence. AB - Herpes simplex virus (HSV) is neurotropic and when inoculated on the mouse footpad is retrogradely transported to the associated dorsal root ganglia (DRG), where infection is established. Previous observations suggest that, after HSV infection, sensory ganglion neurons may mount a sprouting response. In our HSV infected DRG model, we investigate this issue by (1) examining expression of growth-associated protein (GAP-43), a molecule known to be induced by growing axons, and (2) determining ultrastructurally whether HSV-infected dorsal roots contain neurites. In a time course study, we show that GAP-43 is induced both in HSV-infected DRG and their central processes. The increase in GAP-43 is first seen 2 weeks following unilateral footpad inoculation in both cell bodies and dorsal roots, and is sustained at 1 month post inoculation in roots but not in perikarya. Large bundles of unmyelinated small caliber axons, lacking Schwann cell ensheathment, are observed by electron microscopy in dorsal roots 2 weeks and 1 month following inoculation. These profiles resemble developing or regenerating neurites and are rarely seen in roots of mock-infected or uninfected controls. The increased GAP-43 immunoreactivity and ultrastructural changes shown here, in conjunction with previously documented selective neuropeptide and enzyme alterations, confirm that a sprouting response is mounted in sensory ganglia following acute HSV infection. PMID- 9222355 TI - The replicating intermediates of herpes simplex virus type 1 DNA are relatively short. AB - Herpes simplex virus type 1 (HSV-1) replication is thought to occur via a rolling circle type of mechanism, generating large DNA concatemers from which unit length genomes are subsequently cleaved. In this report, we have employed field inversion gel electrophoresis (FIGE), Southern blot hybridization, and endonuclease digestion, to identify and characterize these DNAs. Two species of HSV-1 DNA: (1) genome-length and (2) DNA that remained at the electrophoresis origin (referred to as well-associated DNA) were detected. To ascertain that the latter was large in size and not virion DNA trapped at the origin with high molecular weight cellular DNA, the infected cell DNA was digested with a restriction enzyme that does not cut the viral DNA. In order to do this HSV-1 strain 1702, lacking any XbaI sites in its genome, was utilized. After digestion of samples with XbaI, and FIGE, cellular DNA was seen to migrate into the gel; however, the viral DNA remained in the sample wells. Pulse labeling experiments showed that this large DNA was processed to 150 kb genome lengths. Endonuclease digestion of the well-associated DNA revealed that it contained a greater ratio of joint to terminal fragments than virion DNA-a characteristic of long concatemers. Quantitation of the terminal fragments revealed mainly L termini. Surprisingly, the ratio of joint to terminal fragments was 2.5 suggesting that the lengths of concatemers were short (in the order of 1-2 genome lengths) and that the well association was due to conformation rather than concatemeric length. Because one of these genome lengths is present as the replication intermediate, the growing tail must be less than genome length. Thus genome lengths must be processed from the replication intermediate soon after they are completed. PMID- 9222356 TI - Delayed activation of altered fusion glycoprotein in a chronic measles virus variant that causes subacute sclerosing panencephalitis. AB - We compared the intracellular processing of the fusion (F) glycoproteins of an acute measles virus (MV) Nagahata strain and its relative Biken strain that caused subacute sclerosing panencephalitis (SSPE). Nagahata strain synthesizes a precursor F0 which acquires three asparagine (N)-linked oligosaccharide chains sequentially in 1 h. One oligosaccharide chain on the partially glycosylated F0 is less accessible to endo-beta-N-acetylglucosaminidase H (endo-H) but becomes accessible as the protein becomes fully glycosylated, suggesting a protein conformational change. Biken strain SSPE virus synthesizes a similarly glycosylated F0. However, one oligosaccharide chain on the Biken F0 remains less accessible to endo-H even after the protein is fully glycosylated. The Nagahata F0 is cleaved into the F1 and F2 subunits with a half life of 1 h. The Biken F0 is cleaved with a half life of 4 h. We cloned the F genes of Nagahata and Biken strains and showed by transfection that the defect causing delayed cleavage of F0 resides in the Biken F gene. Sequence analysis predicts a mutation in the cleavage recognition sequence, a truncated carboxyl-terminus, and multiple mutations in F1 of the Biken F protein. Expression of chimeric F genes showed the mutated cleavage recognition sequence and the carboxyl-terminal truncation do not delay cleavage of F0. Instead, delayed F0 cleavage is due to multiple mutations in the extracellular domain of F1, and four amino acid substitutions near the transmembrane region impair endo-H access to the oligosaccharide chain. These results provide detailed information on the normal maturation process of the F protein of MV and additional clues to the mechanisms of MV persistence in the CNS. PMID- 9222357 TI - Capsid protein VP1 deletions in JC virus from two AIDS patients with progressive multifocal leukoencephalopathy. AB - PCR on 52 cerebrospinal fluid (CSF) specimens and 33 brain biopsies obtained from HIV-1 positive patients utilized pairs of primers from both the early region (JTP) and late region (JLP). In these patients, in whom progressive multifocal leukoencephalopathy (PML) was suspected on the basis of clinical symptoms and magnetic resonance imaging (MRI) studies, eight CSFs (15%) and 14 brain biopsy specimens (42%) contained JCV DNA sequences. In two patients' samples, the CSFs were positive for JCV DNA in the VP1 region using the primer pair for the VP1 region (JLP), but the fragment amplified migrated more rapidly than the 129-bp product obtained from prototype JCV(Mad-1) or the fragment amplified from the antigenic variant of JCV known as Mad-11. These patients died 3-4 months after onset of progressive neurological symptoms. Cycle sequencing of the fragments revealed overlapping deletions of 24 and 27 nucleotides. These strains were of different genotypes, designated strain 107 and strain 206. Computer analysis of the VP1 amino acid sequence predicts that the eight or nine amino acid residue deletions represent a surface loop with a high antigenic index. These naturally occurring deletion mutants are the first examples of a phenomenon observed experimentally in the mouse polyoma virus capsid protein VP2. PMID- 9222358 TI - Identification of three new JC virus proteins generated by alternative splicing of the early viral mRNA. AB - The genome of the human polyomavirus JC Virus (JCV) encodes two regulatory proteins, large and small T antigen which are expressed early in a lytic infection, and three structural proteins, VP1, VP2, VP3, which are produced late in an infection. A fourth late protein, agnoprotein, may contribute to the assembly of the virion. In this study, we demonstrate the presence of three additional early proteins, T'135, T'136, and T'165, which are expressed in lytically-infected cells; T'135 is also readily detected in JCV transformants. The three species of T' are phosphoproteins generated via an alternative splicing mechanism. This mechanism involves the excision of a second intron from the large T mRNA using a common donor splice site at JCV nucleotide 4274 and three unique acceptor splice sites at nucleotides 2918, 2777 and 2704 for T'135, T'136 and T'165, respectively. The mutant JCV delta T' was created by converting the G at nucleotide 4274 to an A, thereby disrupting the consensus sequence of the shared donor splice site without altering the amino acid sequence of any early JCV protein. Upon transfection of permissive human brain cells, JCV delta T' replicated its DNA 10-fold less efficiently than did wild type JCV. Passage of extracts of the infected cells on to fresh human brain cells revealed that the expression of T antigen was greatly reduced and the presence of T' proteins was undetectable in the mutant versus wild type JCV-infected cells. PMID- 9222359 TI - Pathogenesis of murine encephalitis limited by defective interfering particles. An immunohistochemical study. AB - To determine whether defective interfering (DI) particles alter viral encephalitis BALB/c mice were inoculated intranasally with standard vesicular stomatitis virus (VSV) and its DI particles. Addition of 10(7) PFU equivalents of DI particles to 10(5) PFU of VSV reduced morbidity but did not delay disease onset. Less mortality was also observed. When 10(3) PFU equivalents of DI particles or UV-irradiated DI particles were substituted, these effects were absent. Attempts to correlate mortality with virus recovered from the brain could not be made due to considerable variations in the few surviving mice. Immunohistochemical analysis obtained from 121 mice showed that inoculation of DI particles limited the specific pathways of VSV antigen dissemination within the central nervous system, and new pathways were not substituted. In the group of mice with reduced mortality due to DI particles, at day 4 post inoculation VSV antigen was limited to the outer layers of the glomeruli of the olfactory bulb and to the accessory olfactory bulb, whereas there was deeper invasion of the olfactory bulb and olfactory ventricular system with mice infected with standard VSV alone. Correlation between mortality and extent of invasion became more difficult to make from 8 days on, when VSV antigens were found in discrete areas of the brain. By 12 days, few surviving mice contained any detectable VSV antigen in their brains. These results demonstrate that DI particles have potential as therapeutic agents. Also, mortality resulting from VSV-induced encephalitis, although poorly understood, may be determined very early, possibly while the virus is replicating at the site of inoculation. PMID- 9222360 TI - Human cytomegalovirus induces IL-6 and TNF alpha from macrophages and microglial cells: possible role in neurotoxicity. AB - Human cytomegalovirus (HCMV) can frequently infect the central nervous system (CNS) in the setting of immunosuppression such as transplantation and infection with the human immunodeficiency virus (HIV). Our laboratory previously reported that HCMV infection of human brain aggregates preferentially infected a microglial/macrophage (M/M) and caused a neuropathology that differed between strains and could occur in the absence of antigen expression. We extended these studies by infecting a human brain cell aggregate model with four low passage clinical isolates of HCMV. Two patterns of cytopathology emerged after infection; a lacy eosinophilic appearance or a glial nodular formation concomitant with a decreased aggregate size. None of the infections were positive for HCMV antigen; however, all were positive for HCMV DNA. We also infected primary macrophages and microglial cells with the same HCMV isolates. Microglial cells were more susceptible to HCMV infection resulting in a lytic infection. Production of potentially neurotoxic cytokines, IL-1, IL-6 and TNF alpha, from HCMV-infected macrophages and microglial cells were evaluated to explain brain aggregate cytopathology. Supernatants from HCMV-infected macrophages and microglial cells produced similar levels of TNF alpha (< 30 pg ml-1) but showed strain and cell source variation in the production of IL-6; microglial cultures produced > 4 fold higher levels. None of the supernatants contained IL-1. Treatment of brain aggregates with either IL-6 or TNF alpha resulted in morphologic alterations and/or a decrease in size consistent with HCMV infection or supernatant treatment. PMID- 9222361 TI - The development of animal model systems for HIV-1 encephalitis and its associated dementia. AB - The human immunodeficiency virus (HIV) is neuroinvasive and can be neurovirulent. Indeed, 20-30% of individuals with the acquired immune deficiency syndrome (AIDS) develop cognitive and motor dysfunction (termed the AIDS dementia complex or HIV dementia) coincident with advanced immunosuppression. Despite massive research efforts to discern viral neuropathogenic mechanisms, much remains incompletely understood. Recently, we and others developed animal model systems to elucidate how HIV infection within the brain can lead to impairment of central nervous system function. In this report, we evaluate each of the published animal models for their ability to mirror HIV dementia. Ease of handling and expense were also under consideration. Ultimately, studies in animal systems should permit a better understanding of the nature of HIV-1-induced neurological injury and aid in the development of effective treatments for this dreaded complication of HIV infection. PMID- 9222362 TI - C58 and AKR mice of all ages develop motor neuron disease after lactate dehydrogenase-elevating virus infection but only if antiviral immune responses are blocked by chemical or genetic means or as a result of old age. AB - Age-dependent poliomyelitis is a paralytic disease of C58 and AKR mice caused by cytocidal infection of anterior horn neurons with neuropathogenic strains of lactate dehydrogenase-elevating virus (LDV). The motor neurons are rendered LDV permissive via an unknown mechanism through the expression of ecotropic murine leukemia virus (MuLV) in central nervous system (CNS) glial cells. Only old mice develop paralytic disease after LDV infection, but mice 5-6 months old or older can be rendered susceptible by suppression of anti-LDV immune responses by a single treatment with cyclophosphamide or X-irradiation before LDV infection. Younger mice appeared to be resistant in spite of this immunosuppresive treatment. The present results confirm that mice as young as 1 month of age possess CNS cells expressing ecotropic MuLV and show that these mice are susceptible to paralytic LDV infection provided their anti-LDV immune responses are blocked for an extended period of time by repeated cyclophosphamide treatments or by a genetic defect. Furthermore, old mice become naturally susceptible to paralytic LDV infection because of an impaired ability to mount a motor neuron protective anti-LDV immune response. PMID- 9222363 TI - Human herpesvirus 6 polymerase chain reaction findings in human immunodeficiency virus associated neurological disease and multiple sclerosis. AB - A role for human herpesvirus 6 (HHV6) in the neurological complications associated with infection by human immunodeficiency virus (neuro-AIDS) and during multiple sclerosis (MS) is not known. For the present study, an improved PCR and immunofluorescence serology method were applied to sera and cerebrospinal fluid (CSF) from 27 neuro-AIDS, 36 MS and 24 non-inflammatory control patients. HHV6 DNA was present in 30-40% of the cellular CSF from all groups. In the acellular CSF, HHV6 could be detected in four of 36 MS, 2 of 27 neuro-AIDS and none of the control patients. HHV6 IgG was present in one of 27 neuro-AIDS, and one of 36 MS patients. HHV6 IgG was present in all patients. There was no correlation between clinical features and HHV6 PCR findings or HHV6 antibodies. The significance of the present documentation of HHV6 DNA in the acellular CSF from a minority of MS and neuro-AIDS patients remains to be determined. PMID- 9222364 TI - Targeting and gene expression in spinal cord motor neurons following intramuscular inoculation of an HSV-1 vector. AB - One problem in devising strategies of gene transfer to the nervous system is targeting specific neuronal populations. To evaluate the potential for using herpes simplex virus (HSV) as a vector for gene transfer to spinal cord motor neurons, the HSV-1 mutant LAT-LTR in which the E. coli beta-galactosidase gene is expressed under control of the HSV LAT core promoter (LAT) and the Moloney murine leukemia virus long terminal repeat (LTR) was inoculated unilaterally into the gastrocnemius muscle. Infectious virus was isolated from the spinal cord on days 3-7 post inoculation (PI). Immunocytochemical labeling of HSV antigen was detected in ipsilateral ventral horn neurons in the spinal cord at day 3 PI and had spread to contiguous spinal cord regions by day 6 PI. No viral antigen was detected at 14 or 28 DPI. beta-galactosidase expression (driven by the LAT-LTR promoter) was detected in neurons of the ventral horn on days 3, 6, 14, and 28 PI. Histological analysis showed mild lesions in the ventral horn on day 3 PI which progressed through days 6, 14 and 28 PI. This study demonstrates the feasibility of gene delivery into spinal motor neurons after injection of an HSV vector at a peripheral muscular site. This approach should prove useful in neurobiological investigations and it suggests a possible application to development of gene therapy for heritable diseases affecting motor neurons. PMID- 9222365 TI - Utility of cerebral proton magnetic resonance spectroscopy in differential diagnosis of HIV-related dementia. AB - Opportunistic infections often coexist with human immunodeficiency virus (HIV) infection in brain. Making the correct diagnosis is often difficult despite recent advances in neuroimaging techniques. 1H magnetic resonance spectroscopy (1H MRS) is an emerging non-invasive examination for diagnosis and monitoring of brain disorders. 1H MRS measures a variety of organic compounds using magnetism and radio waves. Biochemical aberrations in brain, not shown by conventional tests, may be demonstrated by 1H MRS testing. A patient coinfected with HIV and hepatitis B (HBV) presented with progressive dementia. Clinical, neuroradiological and cerebrospinal fluid (CSF) examinations failed to provide a diagnosis in support of either HIV-1-associated cognitive/motor complex or HBV induced hepatic encephalopathy (HE), 1H MRS was used in an attempt to discriminate between these diagnoses. Spectroscopy demonstrated increased glutamine and normal N-acetyl aspartate (NAA) levels, metabolic changes consistent with HE. These findings were later confirmed pathologically. Proton magnetic resonance spectroscopy is a non-invasive test with utility for the differential diagnosis of HIV-associated dementia. PMID- 9222366 TI - Media components influence viral gene expression assays in human fetal astrocyte cultures. AB - In vitro neurovirological studies of viral infectivity or viral gene expression may be confounded by the multiple neural cell types and/or fibroblast contamination present in early passage cultures prepared from dissociated human central nervous system (CNS) tissue. We have developed highly enriched astrocyte cultures for neurovirological study by culturing in a serum-free defined medium, B16, supplemented with basic fibroblast growth factor (FGF-2). Subculture in this medium selects against fibroblast proliferation and favors sustained proliferation of a highly enriched glial fibrillary acidic protein (GFAP) positive cell population. These astrocytes support productive replication of cytomegalovirus (CMV) and transient expression of transfected CMV and human immunodeficiency virus type 1 (HIV-1) viral promoters. By comparison, CNS cultures developed in standard serum-containing medium initially contain predominantly astrocytes, but show increasing contamination with fibroblasts with sequential passage. These cultures support CMV viral synthesis in both fibroblasts and astrocytes, cell types distinguishable only by immunostaining for cell specific antigen. CMV or HIV-1 promoter activities, quantitated by transient gene expression assays, are distinctly lower in CNS cultures maintained in serum containing medium. PMID- 9222367 TI - Frequent mutation in pX region of HTLV-1 is observed in HAM/TSP patients, but is not specifically associated with the central nervous system lesions. AB - Human T-cell leukemia virus type 1 (HTLV-1) is an etiologic agent of adult T-cell leukemia (ATL) and of HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). Recently it has been reported that defective HTLV-1 provirus was detected frequently in the central nervous system (CNS) lesions of HAM/TSP patients. Here we investigated sequence variations of the pX region of HTLV-1 in the CNS and peripheral blood lymphocytes (PBL) of the same patient. The results analyzing 9-13 clones isolated from each specimen indicated that the pX region is highly variable within a patient with HAM/TSP, and the mutations were found at almost random positions within the sequences analyzed. The frequency and pattern of those mutations did not appear to differ significantly between the CNS and PBL of the same patient, although they differed among patients. Similarly, frequent mutations were observed in an asymptomatic carrier of HTLV-1, although the variability was moderate, suggesting that the high variability of the pX sequence is not a specific event in HAM/TSP. However, one asymptomatic carrier showed much less frequent variations very similarly to an ATL patient; both of them harbored clonally expanded infected cells. Thus the apparent low variability was explained by clonal selection of a single species of the provirus by the clonal proliferation of infected cells. These results clearly indicate that mutations including defectives are not specifically associated with the CNS lesions in HAM/TSP patients, but suggest that the random mutations simply reflect the rate of viral replication in individuals and the variants were not inherited frequently. PMID- 9222368 TI - Isolation and characterization of intranuclear ribonucleoprotein complexes associated with double-stranded RNA adenosine deaminase from brain cells: implications for RNA-editing and hypermutation of viral RNA in the CNS. AB - Double-stranded RNA adenosine deaminase (DsRAD), which converts adenosine in duplex RNA to inosine, has been implicated in editing of cellular mRNA and hypermutation of viral RNA in the central nervous system (CNS). We used subcellular fractionation to show that DsRAD in bovine brain tissues is associated with high-molecular-weight ribonucleoprotein (RNP) complexes in the nuclei. DsRAD-associated RNP complexes have apparent molecular mass of up to 500 kDa and buoyant density of 1.35 to 1.42 g cc-1 in CsCl solution. In human glioma cells, DsRAD is also found exclusively in intranuclear RNP complexes that co sediment with the largest RNA species. These DsRAD-associated RNP complexes are dissociated by RNase A or high salt. The RNA component is not essential for DsRAD activity, and the protein component can be separated by dsRNA-affinity column, gel filtration column, and glycerol gradient into enzymatically active protein species with apparent molecular mass ranging from 120 kDa to 70 kDa in polyacrylamide gel. The bovine brain DsRAD has no apparent requirement for low molecular-weight cofactors or metal ions. These results provide insight into the native state of DsRAD in brain cells and have interesting implications for its putative roles in RNA-editing and hypermutation of viral RNA in the CNS. PMID- 9222369 TI - Isolation and characterization of a type II JC virus from a brain biopsy of a patient with PML. AB - Brain tissue of a patient with multiple myeloma suffering from neurological disorders similar to those seen in progressive multifocal leukoencephalopathy (PML) patients was evaluated for the presence of the papovavirus, JCV. Results from polymerase chain reaction (PCR) revealed the presence of JCV with structural organization at the control region which is distinct from well-characterized isolates, ie Mad-1 and Mad-4. The control region of the new isolate, named JCVPhila-1' contains a 22 nucleotide insertion which separates the TATA box from the NF-1 regulatory motif. Only 18 nucleotides of the insert are duplicated in the second copy of the enhancer/promoter of the new isolate, which is 84 nucleotides in size. Results from a transcription assay indicate a modest elevated level of JCVPhila-1 early promoter activity compared to that of JCVMad-4 in glial cell lines. The basal and T-antigen-induced transcriptional activities of the JCVPhila-1 late promoter was lower with respect to Mad-4 late gene activity in glial cells. Of particular interest was the observation that in the cells producing the early protein, T-antigen, JCVPhila-1 DNA replicated more efficiently then the Mad-4 DNA. These results suggest that the alterations seen in the JCVPhila-1 control region may differentially influence early and late gene expression and facilitate amplification of the viral genome in cells derived from the CNS. PMID- 9222370 TI - Amplification of the complete polyomavirus JC genome from brain, cerebrospinal fluid and urine using pre-PCR restriction enzyme digestion. AB - A method is described for amplification of the complete genome of the human polyomavirus JC (JCV) from progressive multifocal leukuencephalopathy (PML) brain, cerebrospinal fluid (CSF) of PML patients, and from the urine of controls without neurological disease. Efficient amplification with the 'long PCR' method is achieved with primers overlapping the single BamHI or EcoRI site following digestion of the DNA sample with the restriction enzyme BamHI or EcoRI, respectively. Cutting of the supercoiled JCV genome allows full separation of the strands during the denaturation step, and permits primer annealing to compete successfully with reassociation of the genomic DNA. With this method a single PCR amplification allows restriction fragment length polymorphism (RFLP) analysis and direct cycle sequencing of any part of the viral genome. RFLP analysis of JCV amplified from CSF has identified a new mutant sequence. Sequencing of clinical samples is useful for typing of JC virus isolates as Type 1 or Type 2 and for characterization of the JCV regulatory region as archetypal or rearranged. PMID- 9222371 TI - Three or more infantile febrile seizures and HHV-6. PMID- 9222372 TI - Coronaviruses. PMID- 9222373 TI - Pseudorabies virus as a neuroanatomical tracer. PMID- 9222374 TI - Quinolinic acid and neurodegeneration in AIDS. PMID- 9222375 TI - Neuropathogenicity of mouse hepatitis virus JHM isolates differing in hemagglutinin-esterase protein expression. AB - The hemagglutinin-esterase (HE) protein of mouse hepatitis virus (MHV) is an optional envelope protein present in only some MHV isolates. Its expression is regulated by the copy number of a UCUAA pentanucleotide sequence present in the leader sequence of the viral genomic RNA. The functional significance of this viral protein so far is not clear. In this report, we compared the neuropathogenicity of two MHV isolates, JHM(2) and JHM(3), which express different amounts of HE protein. Intracerebral inoculation of these two viruses into C57BL/6 mice showed that JHM(2), which expresses an abundant amount of HE protein, was more neurovirulent than JHM(3), which expresses very little HE. Histopathology showed that early in infection, JHM(2) infected primarily neurons, while JHM(3) infected mainly glial cells. JHM(3) eventually infected neurons and caused a delayed death relative to JHM(2)-infected mice, suggesting that the progression of JHM(3) infection in the central nervous system was slower than JHM(2). In vitro infection of JHM(3) in primary mixed glial cell cultures of astrocyte-enriched cultures yielded higher virus titers than JHM(2), mimicking the preferential growth of JHM(3) in glial cells in vivo. These findings suggest that the reduced neuropathogenicity of JHM(3) may correlate with its preferential growth in glial cells. Sequence analysis showed that the S genes of these two viruses are identical, thus ruling out the S gene as the cause of the difference in neuropathogenicity between these two viruses. We conclude that the HE protein contributes to viral neuropathogenicity by influencing either the rate of virus spread, viral cell tropism or both. PMID- 9222376 TI - Phenotypic and functional characterization of CD8+ T lymphocytes from the central nervous system of rats with coronavirus JHM induced demyelinating encephalomyelitis. AB - Intracerebral infection of Lewis (LEW) inbred rats with the neurotropic strain of the murine coronavirus JHM (JHMV) frequently results in a monophasic paralytic disease. In contrast, infection of Brown Norway (BN) inbred rats does not lead to clinical disease. Previous findings indicated that in both rat strains brain infiltrating leukocytes consisted mainly of CD8+ T lymphocytes. Here, we phenotypically as well as functionally characterised this T cell subset after isolation from the central nervous system (CNS). Using JHMV-infected target cells, MHC class I restricted, cytotoxic T lymphocytes were demonstrated to be present in the leukocyte fraction from the CNS of both, susceptible LEW and disease-resistant BN rats. However, compared to infected, but healthy BN rats, diseased LEW rats generated an enhanced cytotoxic immune response which became most prominent at the maximum of neurological disease. Recently published observations from our laboratory demonstrated a strong virus-specific antibody response in the CNS of BN rats. In LEW rats, however, the response was delayed and of low magnitude. This suggests, that consequences of cytotoxic T lymphocyte action in JHMV-infected CNS tissue largely depend on the efficacy of an accompanying virus-specific humoral immune response. PMID- 9222377 TI - Influence of infectious dose upon productive replication and transynaptic passage of pseudorabies virus in rat central nervous system. AB - Pseudorabies virus (PRV) is a neurotropic swine alpha herpesvirus that characteristically invades the nervous system and replicates within synaptically linked populations of neurons. The invasive characteristics and ability of this family of viruses to replicate in neurons of the central nervous system (CNS) have been exploited to map functionally related populations of neurons in a variety of systems. In this report, we examined the effects of strain and concentration on the ability of PRV to infect retinal ganglion cells and pass transneuronally through central visual circuits. We find that the ability of virulent (PRV-Becker) and attenuated (PRV-Bartha) strains of PRV to produce a productive infection of visual circuitry is directly dependent upon the infectious of the injected virus. Injections of at least 10(5) total plaque forming units produce 100% infectivity, whereas lower infectious doses substantially reduce the percentage of animals exhibiting productive infection via this route of inoculation. Furthermore, the virulent strain of PRV consistently infects a higher percentage of animals across a broader range of titers than attenuated virus. These data demonstrate that viral titer and strain are important variables that should be considered in the design of studies and interpretation of data derived from investigations employing this pathogen for circuit analysis. PMID- 9222378 TI - Dendritic morphology of cardiac related medullary neurons defined by circuit specific infection by a recombinant pseudorabies virus expressing beta galactosidase. AB - The transneuronal herpesvirus tracer, pseudorabies virus (PRV) was used to determine the dendritic architecture of cardiac-related neurons. We constructed a derivative of the Bartha strain of PRV called PRV-BaBlu, that carries the lacZ gene of E. coli. Expression of beta-galactosidase by this recombinant virus enabled us to define the dendritic morphology of motoneurons and interneurons that innervate the heart. beta-galactosidase antigen filled dendritic processes that were clearly revealed by antibodies to beta-galactosidase. In contrast, the standard enzymatic reaction for detection of beta-galactosidase activity stained the cell soma well, but was inferior for labeling dendrites. Following PRV-BaBlu cardiac injection, infected neurons were clearly defined and labeled dendrites could be traced for long distances, sometimes greater than 800 microns from the cell body. Labeled dendrites of cardiomotor neurons primarily located in the nucleus ambiguus (NA) were extensive and sometimes intertwined with dendrites from other labeled motoneurons. Dendrites of labeled neurons in the dorsal motor nucleus of the vagus (DMV) typically extended in the mediolateral direction in the transverse plane. Transynaptically labeled interneurons interposed between the cardiorespiratory region of the nucleus tractus solitarius (NTS) and the NA were primarily located in the NA region and the reticular arc, the area between the DMV and NA. These interneurons had long dendrites extending along the reticular arc in the transverse plane. The dendritic arborizations of infected cardiac-related neurons in the NTS were variable in extent. We conclude that antibody detection of beta-galactosidase expressed by PRV-BaBlu after infection of neural cardiac circuits provides a superior method to define the dendrites and dendritic fields of cardiac-related motoneurons and interneurons. PMID- 9222379 TI - Quinolinic acid production is related to macrophage tropic isolates of HIV-1. AB - We sought to determine whether the neurotoxin quinolinic acid (QUIN) was produced by macrophages or lymphocytes infected with isolates of HIV-1 with varying degrees of macrophage tropism derived from patients with varying stages of AIDS dementia complex (ADC). Highly macrophage tropic isolates and minimally macrophage tropic isolates were used to inoculate macrophages and QUIN production was measured. Similarly, QUIN production from macrophages was monitored using a purified cell free highly macrophage tropic isolate and laboratory isolates SF33 and SF2. Each of these experiments was also performed with lymphocytes. We found that macrophages infected with macrophage tropic isolates of HIV-1 led to QUIN production while lymphocytes did not produce QUIN. The ability of the HIV-1 infected macrophages to produce QUIN was related to the viral inoculum and the degree of macrophage tropism of the isolate. The severity of ADC in the patient from whom a particular isolate was derived was not per se a determining factor for QUIN production. Purified cell free ADC isolates also led to QUIN production by macrophages thereby suggesting that HIV-1 infection alone is capable of inducing QUIN production. PMID- 9222380 TI - Neurocytotoxity of quinolinic acid in human brain cultures. AB - Quinolinic acid (QUIN) is a tryptophan metabolite which has been found to be an excitotoxin in rats, although its toxicity in humans is unknown. QUIN has been implicated in the pathogenesis of AIDS dementia complex. This study examined the effect of QUIN on human fetal brain tissue in vitro. After at least 14 days in vitro, QUIN was added to the cultures in the feeding medium, and lactate dehydrogenase (LDH) efflux at 20 h and neuronal morphology were used as a measure of neuronal injury. LDH levels in media from cultures exposed to QUIN concentrations of 5 and 10 mM were consistently elevated compared to controls. Overall, LDH levels in media from cultures exposed to lower QUIN concentrations did not differ significantly from controls. These data are comparable to animal in vitro studies, and support the hypothesis that QUIN is toxic to human central nervous system neurons and further strengthen its potential role in the pathogenesis of AIDS dementia complex. PMID- 9222382 TI - Interaction of 68-kDa TAR RNA-binding protein and other cellular proteins with prion protein-RNA stem-loop. AB - The RNA stem-loop structure of the trans-activating region TAR sequence of human immunodeficiency virus-1 mRNA is the binding site for a number of host cell proteins. A virtually identical set of proteins from HeLa nuclear extracts was found to bind to the predicted RNA hairpin element of prion protein (PrP) mRNA, as demonstrated in UV cross-linking/RNase protection and Northwestern assays. We show that the cellular TAR loop-binding protein, p68, is among those proteins which associate with PrP RNA. Competition experiments with various TAR RNA mutants revealed that binding of partially purified p68 to PrP RNA stem-loop occurs sequence-specifically. The 100-kDa 2-5A synthetase which is involved in the cellular antiviral defense was able to bind to PrP mRNA stem-loop in Northwestern blots with cytosolic proteins from HeLa cells treated with interferon. However, the PrP RNA failed to activate this enzyme in vitro, in contrast to TAR RNA. PMID- 9222381 TI - Molecular mimicry between HIV-1 gp41 and an astrocyte isoform of alpha-actinin. AB - A 100-kDa astrocyte antigen previously shown to cross-react with a monoclonal antibody (MAb) generated against amino acids (aa) 598 to 609 of the transmembrane protein gp41 of human immunodeficiency virus type 1 [HIV-1] has now been molecularly characterized and found to be an alpha-actinin (alpha-actinin) related protein. Western blot analyses of human astrocytoma cells fractionated by differential centrifugation and detergent phase separation showed that the antigen was membrane associated. The astrocyte protein was purified to apparent homogeneity by immunoaffinity chromatography. Amino acid analysis of three peptide fragments obtained by cleavage of the purified 100-kDa protein revealed sequence identities of 77, 83 and 100% to a non-muscle isoform of human alpha actinin. In addition, the aa 598-609 sequence of gp41 recognized by MAb 781.4, and the aa 581-597 sequence recognized by another cross-reactive MAb 781.3, were 73% and 53% similar to regions of alpha-actinin. This molecular mimicry between gp41 and alpha-actinin was supported by antibody cross-reactivity in Western immunoblot and ELISA analyses. Both anti-gp41 and anti-alpha-actinin MAbs bind to the surface of the human astrocytoma cells as detected by a cell surface binding assay and immunofluorescence. Antibodies made against this immunodominant region of gp41 in the serum and CSF of HIV-infected individuals have access to astrocytes within the CNS. The identification of the astrocyte antigen as an alpha-actinin related protein will allow further work to determine how this immunological cross-reactivity could perturb astrocyte function and contribute to HIV neuropathology. PMID- 9222383 TI - Attempted modulation of herpes simplex virus (HSV) infection of neurons in culture by fibroblast growth factor. AB - It has been suggested that fibroblast growth factor (FGF) receptors may mediate entry of herpes simplex virus (HSV) to susceptible cells. We investigated the possible modulation of acute lytic HSV infection of cultured rat neurons by basic FGF, using cell-specific markers and indirect immunostaining. Dissociated neural cell cultures were prepared from the medial septal region of the basal forebrain, the hippocampus and dorsal root ganglion (DRG). The proportion of neurons in the cultures was enhanced by the addition of cytosine arabinoside (2 microM) in the hippocampal and DRG cultures and by inversion of the coverslips in septal cultures. The percentage of MAP2+ and neurofilament+ neurons in these cultures varied between 9 and 95%. Cultures were treated with basic FGF (4-5 ng ml-1) continuously and infected with wild-type HSV-1, untreated uninfected cultures acting as controls. Both FGF binding and FGF receptors were demonstrated in hippocampal, and to a much lesser extent, DRG neurons. In repeated experiments it was found that FGF treatment did not have a significant effect on lytic infection in any of the neuronal populations studied as assessed by the development of viral antigen expression and comparison of identified neurons in FGF, treated versus untreated, infected cultures. Our data show that FGF does not have a 'neuroprotective' effect on HSV infection of either central or peripheral neurons. PMID- 9222384 TI - Protective effects of interferon-gamma in intraocular herpes simplex type 1 infection do not depend on major histocompatibility complex class I or class II expression. AB - Intraocular infection with herpes simplex virus type I strain F (HSV-1) induces bilateral retinitis, the expression of both MHC class I and II molecules and activation of CD4 and CD8 cells. To investigate the role of MHC upregulation in IFN-gamma mediated antiviral effects in intraocular infection with HSV-1, we infected MHC deficient mice and mice with an additional ectopic site of IFN-gamma production in their retina (rho gamma) intravitreally with HSV-1 into one eye. Protective effects of IFN-gamma in intraocular HSV-1 infection were notable as sparing of the contralateral non-inoculated eye from retinitis, and were not dependent on MHC class I and class II expression, thus limiting the importance of MHC expression for the outcome of viral infection in vivo. PMID- 9222385 TI - Delayed activation of altered fusion glycoprotein in a chronic measles virus variant that causes subacute sclerosing panencephalitis. AB - We compared the intracellular processing of the fusion (F) glycoproteins of an acute measles virus (MV) Nagahata strain and its relative Biken strain that caused subacute sclerosing panencephalitis (SSPE), Nagahata strain synthesizes a precursor F0 which acquires three asparagine (N)-linked oligosaccharide chains sequentially in 1 h. One oligosaccharide chain on the partially glycosylated F0 is less accessible to endo-beta-N-acetylglucosaminidase H (endo-H) but becomes accessible as the protein becomes fully glycosylated, suggesting a protein conformational change. Biken strain SSPE virus synthesizes a similarly glycosylated F0. However, one oligosaccharide chain on the Biken F0 remains less accessible to endo-H even after the protein is fully glycosylated. The Nagahata F0 is cleaved into the F1 and F2 subunits with a half life of 1 h. The Biken F0 is cleaved with a half life of 4 h. We cloned the F genes of Nagahata and Biken strains and showed by transfection that the defect causing delayed cleavage of F0 resides in the Biken F gene. Sequence analysis predicts a mutation in the cleavage recognition sequence, a truncated carboxyl-terminus, and multiple mutations in F1 of the Biken F protein. Expression of chimeric F genes showed the mutated cleavage recognition sequence and the carboxyl-terminal truncation do not delay cleavage of F0. Instead, delayed F0 cleavage is due to multiple mutations in the extracellular domain of F1, and four amino acid substitutions near the transmembrane region impair endo-H access to the oligosaccharide chain. These results provide detailed information on the normal maturation process of the F protein of MV and additional clues to the mechanisms of MV persistence in the CNS. PMID- 9222387 TI - Lysine clonixinate in the treatment of primary dysmenorrhea. AB - The efficacy and tolerance of Lysine Clonixinate (LC), a NSAID with prostaglandin synthesis inhibiting mechanism was studied in 24 patients with primary dysmenorrhea according to a double-blind randomized crossover Placebo (P) controlled design with patients serving as their own controls. Treatment consisted in administering 1 tablet of LC or P q6h as from onset of menstrual pain during 5 days and 6 menstrual cycles. Patients were controlled monthly as from the 5th day of the cycle, rating changes in pain intensity according to a 4 point scale, presence of pain during pre-, post- and menstrual periods; possible intracycle changes, amount of bleeding, tolerance and related total and general signs and symptoms. Intensity of baseline menstrual pain amounted to 2.9. Menstrual, intramenstrual and postmenstrual pains were observed in 19 out of 24, 24/24 and only 2 out of the 24 patients, respectively. Concomitant symptoms consisted in headache (12), mastalgia (14) and discomfort (12). Results were obtained by averaging the data from the treatment periods with each drug. Menstrual pain was reduced from 2.9 +/- 0.7 to 1.9 +/- 0.7 with P administration and to 0.66 +/- 0.4 with the administration of LC, a highly significant difference between treatments (p < 0.0001). Premenstrual pain was reduced nonsignificantly from 0.79% to 0.58% with P administration and significantly to 0.29% with administration of LC (p < 0.001). Intramenstrual pain affecting all patients at baseline was reduced significantly by 9% with P and also significantly by 50% with LC (p < 0.001). No differences were encountered in concomitant symptoms during P treatment periods while the incidence was significantly reduced with LC (p < 0.0001). No changes in cycle duration or amount of bleeding were observed between treatments. No adverse events were reported. PMID- 9222386 TI - Polyamines in the male reproductive system. AB - This review covers some common aspects of the biosynthesis, interconversion pathways and biochemical functions of polyamines. A particular emphasis is given in experimental models as well as humans, to their presence in the male gonad, prostate gland, seminal vesicles, epididymis and semen. The interaction between hormones (androgens, LH, FSH and PRL) and the main enzymes involved on the polyamine biosynthesis, and the relationship of these compounds on cell growth and differentiation, are also discussed. In this regard, an attention is offered to the potential role of polyamines during early spermatogenesis stages and the use of some enzymes involved in their biosynthesis as sensitive and specific markers of the action of androgens and antiandrogens in the epididymis. Finally, a special issue is addressed to the controversial information documented on polyamines, their oxidation products and the relationship with male fertility. PMID- 9222388 TI - [Edema and myonecrosis induced by Bothrops jararaca venom of Argentina in mice]. AB - Myonecrotic and oedema-inducing activities of Bothrops jararaca of Argentina were studied. For oedema-inducing activity 0.05 ml of different solutions of venom in 0.9% NaCl were injected in mice. The dose of 0.86 micrograms/20 g mouse induced an oedema of 30% respect the other member in one hour. The myonecrotic effects were studied injecting mice gastrocnemius muscle with 70 micrograms of venom in 0.1 ml of 0.9% NaCl, which were sacrificed in different time. The inoculation area was obtained for hystophatological process. Animal sacrificed 30 minutes after the inoculation showed oedema, hemorrhage and inflammatory infiltrate. Those sacrificed in one and three hours after the inoculation also had got necrosis. PMID- 9222389 TI - Bioavailability of microencapsulated ferrous sulfate in fluid milk studies in human beings. AB - Iron deficiency is one of the most important nutritional problems in the world. The best method to overcome this problem is the fortification of foods with highly bioavailable iron. Fluid milk is a massive consumption food with an easy access and which is generally the only food intake during the first months of life. Therefore the fortification of fluid milk with highly bioavailable iron and no detectable alterations of its sensorial characteristics was studied in the present work. This procedure was made possible using a new type of ferrous sulfate, stabilized and microencapsulated with soy lecitin (SFE-171). The iron concentration of the fortified milk is 12 mg per liter. In order to study the iron absorption from milk fortified with this product, SFE-171 was labeled with 59Fe and given to 29 volunteers with a normal iron status, each of which received an iron quantity of 3 mg in 250 ml of fluid milk. The average iron absorption was (10.2 +/- 4.7) %. This result shows that the iron given in this physicochemical form has the advantage of a high bioavailability and it is possible that this product will be the first attempt for an adequate solution of iron deficiency. PMID- 9222390 TI - Inhibitory effects of diazepam in rat vas deferens: role of calcium. AB - Diazepam and Ro5-4864 effects on noradrenaline-induced rat vas deferens contractions were studied. We investigated whether central or peripheral type benzodiazepine receptors were involved, by studying the effects of the selective central or peripheral benzodiazepine receptor antagonists, flumazenil (Ro 151788) or PK 11195 respectively. Diazepam interactions with GABA, adenosine, theophylline, and hypercalcic medium (3.5 mM) were studied. Also, we investigate diazepam effect on KCl depolarized vas deferens. Results showed that diazepam (10(-4) to 1.7 x 10 (-4) M) and Ro 5-4864 (10(-5) to 5.5 x 10(-5) M) inhibited NA induced vas deferens contractions and that neither flumazenil nor PK 11195 antagonized diazepam or Ro 5-4864 inhibitory effects respectively. GABA, adenosine and theophylline did not modify neither NA vas deferens response nor diazepam inhibitory action. Diazepam effect was significantly reduced in and 3.5 mM calcium medium and KCl vas deferens response was inhibited by diazepam 1.3 x 10(-5) and 1.3 x 10(-4) M. It is concluded that in rat vas deferens diazepam effect seems to be related with calcium mobilization. PMID- 9222391 TI - [Tumors caused by methylcholanthrene and lapachol. Follow-up of development with cytology]. AB - This present work was carried out in order to study the effect of Lapachol (LAP) administrated to rats simultaneously with a chemical carcinogen 20 Methylcholanthrene (MCA). Animals were divided in 4 groups: A-Group treated with 80 mg of MCA I(n = 11 rats), B-Group treated with 80 mg of MCA+LAP 100 mg/kg weight/day, (n = 15 rats), C-Group treated with LAP 100 mg/kg weight/day (n = 12 rats), D-Group control-no treatment (n = 13 rats). Cytological studies as well as cytochemical techniques allowed the recognition of benign and malignant conditions at the time of the tumor appearance. Histopathological evaluation posteriorly confirmed the development of tumors in 53% of the animals in group B. Morphologically consistent with poorly differentiated adenocarcinomas of the salivary glands and fibroadenomas of the breast in 18.2% (2/11) of the rats in group A. Besides the presence of one or several supra-hepatic nodules in vecinity of the suspensory ligament corresponding to nodular hyperplasia were observed in 33% (4/12) of group C and in 13.3% (12/15) of group B. No nodules were observed in groups A and D. Tubular dilatation of kidneys were noted in 60% (9/15) and 83.3/ (10/12) of the rats in group E and C respectively. From the original salivary gland tumor a series of transplantables tumors were developed and followed by cytological evaluations. The importance of the cytological and histopathological diagnosis for the pharmacological effects studies of certain drugs like Lapachol that are widely used as antitumoral agents without exact knowledge of the adverse effects is emphasized. PMID- 9222392 TI - Porphyrinogen carboxylyase. Studies on the existence of isoenzymes. AB - Porphyrinogen carboxylyase from normal rat liver was subjected to purification methods. Two different purification protocols were used. In both cases, the initial steps consisted in obtaining a liver homogenate, followed by centrifugation, salt precipitation and phosphate gel absorption. Scheme I consisted in then submitted the preparation to DEAE-cellulose, followed by Sephacryl S-200 and Phenyl-sepharose sequential column chromatographies. Scheme II involved an affinity column followed by a Sephadex G-75 gel filtration column. In both cases, the enzyme was stored at -20 degrees C until its assay. The addition of 2mM dithiotreytol to the incubation media or to the enzyme extract before storage, did not help improve the activity nor the stability of the enzyme. Those fractions containing the maximal enzyme activity, detected using Uroporphyrinogen III or Pentacarboxy-porphyrinogen III as substrate, were not always present in the same tubes for the different columns employed. In addition, the degree of purification obtained in some steps was different according to the substrate employed. The results suggest the existence of at least two isoenzymes for rat liver porphyrinogen carboxy-lyase. PMID- 9222393 TI - Prazosin and stress effect on tumoral growth of 7,12-dimethylbenz[A]anthracene induced rat mammary tumors. AB - Repeated isolation stress and prazosin effect were evaluated in 7,12 dimetylbenz[A]anthracene (DMBA) mammary tumors. Tumor volume was significantly lower in stressed than in control animals from 10 to 52 days considering day 1 the moment when tumors became palpable and treatment began. Control Prazosin (0.5 mg/kg) rats showed diminished tumor volume after 40 days. Stress Prazosin curve was similar to stress alone. The proportion of progressing tumors in control was significantly higher than in stressed groups, regardless of Prazosin administration. Body weight gain was similar in every group throughout the experiment. Behavioral studies were performed when stress effect was no longer evident. Grooming and the number of fecal boli were similar in all groups, as well as prolactin serum levels, suggesting that habituation took place. No significant differences were observed between groups for estrogen receptors. However, a greater concentration of progesterone receptors was found in Stressed rats, compared to all other groups. We conclude that the decrease of tumor volume provoked by stress could not be reversed by the alpha 1-adrenergic antagonist prazosin. Then, it appears that the main effect of stress is not mediated by the alpha 1-adrenergic receptors. Higher progesterone receptors in stressed rats could explain the differences observed. PMID- 9222394 TI - The rat thymus contains a heparin-binding factor that modulates steroidogenesis in the testis. AB - Thymus development and function are under the influence of hormones secreted by the gonads and pituitary. On the other hand, thymus is crucial for the development of reproductive capacities in female and male rats and we have shown that a factor derived from the prepubertal rat thymus has antigonadotropic effect in ovarian and testis cells in vitro. In the present paper we show that the rat thymic factor which modulates gonadotropin action in the gonads is an heparin binding factor. This capacity was also used as a useful tool to obtain this activity from semipure extracts. An acetone extract was prepared from 15 day old male rats and subjected to molecular filtration chromatography. The activity, of those fractions was investigated in a testis cells bioassay, by measuring testosterone secretion under basal and hCG-stimulation. Active fraction were processed in an heparin-Sepharose affinity column. We found that fractions that eluted with 0.6 and 2M NaCl/10mM Tris had biological specific activity. The electrophoretic procedure showed that the apparent molecular weight of the Heparin Sephadex binding factor is 60 kDa. Since this factor was obtained from a protein peak that eluted in the volume of carbonic anhidrase a dimerization process could be involved. Present results show that the rat thymus has an heparin-binding factor that interacts with hCG in testis cells. This factor could play an interesting role in the mutual influence between thymus and gonads. PMID- 9222395 TI - [Effect of N-Nitro-L-Arginine on the concentration and relaxation of vascular smooth muscle]. AB - The arterial smooth muscle contractility is regulated by the free intracellular calcium concentration and by alpha 1 and alpha 2 adrenergic receptor stimulation (norepinephrine (NE), phenylephrine (PHE) and clonidine), inducing contraction, increase in vascular tension and increase in intracellular calcium concentration. This response is increased in the absence of endothelium. On the other hand, the relaxation of isolated arteries induced by acetylcholine (ACh) is endothelium dependent and modulate by EDRF. We have studied the effect of pre incubation with N-nitro-L-arginine (L-NA) on the alpha-adrenergic-induced-contraction and ACh induced relaxation, respectively in isolated rat toraxic aorta strips in comparison with the effect produced by 70 mM KCl which is taken as control. The results show a significant difference (p < 0.001) in the presence of L-NA on the PHE-induced contraction compared to the absence of L-NA. At the three ACh concentration used, the presence of L-NA induced relaxation less than 20%. The absence of L-NA produced 100% relaxation returning to base line. L-NA induces an increase in calcium entry through receptor modulated calcium channels, and because the inhibition of the synthesis of EDRF it would produce an increase in vascular tension. In the same way, relaxation is explained by ACh-stimulated EDRF release, which is inhibited by L-NA. Considering the vascular endothelium as a paracrine organ, is important and interesting to study the pharmacomodulation of the NO/EDRF effects and, for the same reason, to consider L-NA in the pharmacologic arsenal. PMID- 9222396 TI - Glutamatergic transmission in the nucleus tractus solitarii: from server to peripherals in the cardiovascular information superhighway. AB - Afferent nerves carrying signals from mechanoreceptors in the aortic arch and carotid sinus terminate predominantly in the nucleus tractus solitarii (NTS). Signal transduction and neurotransmission in the NTS are critical for central cardiovascular reflect control, but little was known about either until the late 1970's. None of the numerous neuroactive chemicals found in the NTS had met strict criteria as a neurotransmitter in the baroreflex arc until data suggested that the excitatory amino acid L-glutamate (GLU) might be released from baroreceptor afferent terminals in the NTS. In anesthetized animals microinjection into the NTS of GLU, which can be demonstrated in terminals in the NTS, produces cardiovascular responses like those seen with activation of the baroreceptor reflex. Similar responses occur in awake animals if the chemoreceptor reflex is eliminated; otherwise, in conscious animals responses mimic those of chemoreceptor reflect activation. GLU released in the NTS upon selective activation of the baroreceptor, and possibly the chemoreceptor, reflex. Responses to selective agonists as well as baroreflex responses are eliminated by GLU antagonists microinjected into the NTS. Non-NMDA (N-methyl-D-aspartic acid) receptors seem to predominate at primary baroreceptor synapses in the NTS while NMDA receptors may be involved at later synapses. Although inhibition of soluble guanylate cyclase attenuates responses to ionotropic glutamate agonists in the NTS, nitric oxide does not seem to play a role in glutamate transmission in the NTS. GLU may also participate in transmission at cardiovascular neurons beyond the NTS. For example, a role has been suggested for GLU in the ventrolateral medulla and spinal cord. Work continues concerning GLU signal transduction and mechanisms that modulate that transduction both at the NTS and at other cardiovascular nuclei. PMID- 9222397 TI - Anhydrobiosis in yeast: activation effect. AB - Intracellular substances released into the medium during rehydration of dry yeast cells can significantly improve the quality of a synthetic medium. Acceleration of yeast growth in this medium and increased yield of biomass are observed simultaneously. The change in the molecular arrangement of intracellular membranes as a result of the strong dehydration of live organisms is a negative phenomenon that reduces the level of cell viability. However, this phenomenon also represents an adaptive mechanisms which facilitates the maintenance of population viability as a whole under extreme environmental conditions. PMID- 9222398 TI - Optimized Fast-FISH with alpha-satellite probes: acceleration by microwave activation. AB - It has been shown for several DNA probes that the recently introduced Fast-FISH (fluorescence in situ hybridization) technique is well suited for quantitative microscopy. For highly repetitive DNA probes the hybridization (renaturation) time and the number of subsequent washing steps were reduced considerably by omitting denaturing chemical agents (e.g., formamide). The appropriate hybridization temperature and time allow a clear discrimination between major and minor binding sites by quantitative fluorescence microscopy. The well-defined physical conditions for hybridization permit automatization of the procedure, e.g., by programmable thermal cycler. Here, we present optimized conditions for a commercially available X-specific alpha-satellite probe. Highly fluorescent major binding sites were obtained for 74 degrees C hybridization temperature and 60 min hybridization time. They were clearly discriminated from some low fluorescent minor binding sites on metaphase chromosomes as well as in interphase cell nuclei. On average, a total of 3.43 +/- 1.59 binding sites were measured in metaphase spreads, and 2.69 +/- 1.00 in interphase nuclei. Microwave activation for denaturation and hybridization was tested to accelerate the procedure. The slides with the target material and the hybridization buffer were placed in a standard microwave oven. After denaturation for 20 sec at 900 W, hybridization was performed for 4 min. at 90 W. The suitability of a microwave oven for Fast FISH was confirmed by the application to a chromosome 1-specific alpha-satellite probe. In this case, denaturation was performed at 630 W for 60 sec and hybridization at 90 W for 5 min. In all cases, the results were analyzed quantitatively and compared to the results obtained by Fast-FISH. The major binding sites were clearly discriminated by their brightness. PMID- 9222399 TI - Single-step purification of crotapotin and crotactine from Crotalus durissus terrificus venom using preparative isoelectric focusing. AB - We describe the isolation of crotoxin, a presynaptic B-neurotoxin, as well as its subunits B (crotactine) and A (crotapotin) from lyophilized Crotalus durissus terrificus venom by a single-step preparative isoelectric focusing procedure. From 98 mg of dried venom protein 20.1 mg of crotactine and 13.1 mg of crotapotin were recovered in the first step of focalization and 4.2 mg in a second run. These values correspond to 35.7% of the total venom protein applied. Crotactine separated in the 9.3-7.0 pH range (tubes 1-6) and crotapotin in the 1.8-2.8 pH range (tubes 15-19) and both were homogeneous by SDS-PAGE and N-terminal amino acid analysis. Crotactine, a 12-kDA protein, presented hemolytic and phospholipase A2 activity. Thus, using isoelectric focusing we simultaneously purified both toxins in high yields. This method can be used as an alternative for the purification and characterization of proteins from other snake venoms under conditions in which biological activity is retained. PMID- 9222400 TI - Complications of surgical treatment of cervical carcinoma. AB - A total of 302 patients with stage Ib and IIa cervical carcinoma were submitted to radical hysterectomy and lymphadenectomy during the period from 1980 to 1994. The morbidity rate was 37.5% and the mortality rate 0.6%. The most common intraoperative complications were injuries to the great pelvic vessels and the most frequent postoperative complications involved the urinary tract. The leading causes of morbidity were urinary infection (20.8%), bladder dysfunction (9.2%) and ureteral fistulas (2.9%). Although the rate of complications was high, morbidity has been decreasing over the last five years. Thus, radical hysterectomy continues to be one of the methods for the treatment of early cervical carcinoma that presents an acceptable 5-year survival rate. PMID- 9222401 TI - Changes in the electrophysiological parameters of the posterior intestine of Anguilla anguilla (Pisces) induced by oxytocin, urotensin II and aldosterone. AB - In view of the importance of the intestine in the osmoregulation of freshwater fishes, we determined the effects of oxytocin, urotensin II (UII), and aldosterone added to the serosal side of the isolated posterior intestine of the freshwater-adapted teleost Anguilla anguilla on electrophysiological parameters. Oxytocin decreased the short-circuit current (SCC) and transepithelial potential difference (TPD) at concentrations of 1 and 10 mU/ml (to 50% and 42% of control values, respectively), but did not alter these parameters at a concentration of 0.1 mU/ml. UII reduced SCC and TPD at concentrations of 10 nM, 50 nM and 100 nM (to 85% of control values), but increased these parameters at the concentration of 500 nM (to 115% of control values). Aldosterone did not alter SCC or TPD at the concentrations tested (10 nM and 100 nM). Oxytocin may open Na+ channels in the apical membrane, allowing the flow of Na+ to the serosa, reducing SCC and TPD. Should this hypothesis be correct, oxytocin would be important for freshwater adaptation, since it would increase Na+ absorption. The reduction of SCC and TPD in the posterior intestine of A. anguilla induced by UII is evidenced that this neurohormone is also important for freshwater adaptation in teleosts. Aldosterone did not show this effect probably due to the lack of receptors in this organ. PMID- 9222402 TI - A mutant cell line partially responsive to both IFN-alpha and IFN-gamma. AB - A recessive mutant cell line, B7, which is partially responsive to both interferon (IFN)-alpha and IFN-gamma is described. B7 was FACS sorted from a cellular pool, which was obtained from the parental cell line 2C4, after several rounds of mutagenesis. The partial responsiveness to IFN was observed both in terms of expression of cell surface markers (CD2, class I and II HLAs) and mRNA expression of IFN-stimulated genes (9-27; 6-16; 2'-5' OAS; GBP and HLA-DR alpha). A genetic cross with the U4 mutant (JAK1-, a member of the Janus family of nonreceptor tyrosine kinase) did not restore full IFN responsiveness to B7, and JAK1 cDNA transfection into B7 restored the wild phenotype of the cell line, defining B7 as a member of the U4 complementation group. Nevertheless, JAK1 mRNA was not detected in this mutant. Transcriptional regulator complexes such as IRF1/2 (IFN-regulatory factor) and ISGF3-gamma (IFN-stimulated gene factor) were constitutively formed in the B7 mutant and co-migrated with the IFN-induced complexes expressed in the parental cell line 2C4. Thus, this cell line seems to be useful for understanding cis-acting elements governing JAK1 mRNA expression. PMID- 9222403 TI - Association of leprosy with HLA-DR2 in a Southern Brazilian population. AB - The association between HLA specificities and leprosy was investigated in a southern Brazilian population. One hundred and twenty-one patients and 147 controls were typed for HLA-A, B, Cw, DR and DQ. Patients were subdivided into the following subgroups, according to clinical, histological and immunological criteria: lepromatous (N = 55), tuberculoid (N = 32), dimorphous (N = 20), and indeterminate (N = 14). The frequencies of HLA specificities were compared between the total group of patients and controls, and between the same controls and each subgroup of patients. After correction of the probabilities, deviations, were not significant, except for the DR2 specificity, which presented a frequency of 44.2% in the total group of patients and 56.3% in the subgroup of individuals with the tuberculoid form of the disease, compared to 23.3% in the controls. Stratified analysis showed that the increased DR2 frequency in the total group of patients was due to the subgroups with tuberculoid and dimorphous forms. The relative risk of tuberculoid leprosy for DR2-positive individuals was 4.2, and the etiologic fraction of DR2 was 0.429. In conclusion, a positive association of the DR2 specificity with the tuberculoid form of leprosy, but not with the lepromatous, dimorphous, or indeterminate forms, was demonstrated in this Southern Brazilian population. PMID- 9222404 TI - Behavioral effects of "vehicle" microinjected into the dorsal periaqueductal grey of rats tested in the elevated plus maze. AB - To investigate the behavioral effects of different vehicles microinjected into the dorsal periaqueductal grey (DPAG) of male Wistar rats, weighing 200-250 g, tested in the elevated plus maze, animals were implanted with cannulas aimed at this structure. One week after surgery the animals received microinjections into the DPAG of 0.9% (w/v) saline, 10% (v/v) dimethyl sulfoxide (DMSO), 2% (v/v) Tween-80, 10% (v/v) propylene glycol, or synthetic cerebrospinal fluid (CSF). Ten min after the injection (0.5 microliters) the animals (N = 8-13/group) were submitted to the elevated plus maze test. DMSO significantly increased the number of entries into both the open and enclosed arms when compared to 0.9% saline (2.7 +/- 0.8 and 8.7 +/- 1.3 vs 0.8 +/- 0.3 and 5.1 +/- 0.9, respectively. Duncan test, P < 0.05), and tended to increase enclosed arm entries as compared to 2% Tween-80 (8.7 +/- 1.3 vs 5.7 +/- 0.9, Duncan test, P < 0.10). In a second experiment no difference in plus maze exploration was found between 0.9% saline or sham-injected animals (N = 11-13/group). These results indicate that intra DPAG injection of some commonly used vehicles such as DMSO, saline or Tween-80 affects the exploratory activity of rats exposed to the elevated plus maze in statistically different manners. PMID- 9222405 TI - Correlations between ANP concentrations in atria, plasma and cerebral structures and sodium chloride preference in Wistar rats. AB - We determined whether ANP (atrial natriuretic peptide) concentrations, measured by radioimmunoassay, in the ANPergic cerebral regions involved in regulation of sodium intake and excretion and pituitary glad correlated with differences in sodium preference among 40 Wistar male rats (180-220 g). Sodium preference was measured as mean spontaneous ingestion of 1.5% NaCl solution during a test period of 12 days. The relevant tissues included the olfactory bulb (OB), the posterior and anterior lobes of the pituitary gland (PP and AP, respectively), the median eminence (ME), the medial basal hypothalamus (MBH), and the region anteroventral to the third ventricle (AV3V). We also measured ANP content in the right (RA) and left atrium (LA) and plasma. The concentrations of ANP in the OB and the AP were correlated with sodium ingestion during the preceding 24 h, since an increase of ANP in these structures was associated with a reduced ingestion and vice-versa (OB: r = -0.3649, P < 0.05; AP: r = -0.3291, P < 0.05). Moreover, the AP exhibited a correlation between ANP concentration and mean NaCl intake (r = 0.4165, P < 0.05), but this was not the case for the OB (r = 0.2422). This suggests that differences in sodium preference among individual male rats can be related to variations of AP ANP level. Earlier studies indicated that the OB is involved in the control of NaCl ingestion. Our data suggests that the OB ANP level may play a role mainly in day-to-day variations of sodium ingestion in the individual rat. PMID- 9222406 TI - Ethanol-induced hypothermia in rats is antagonized by dexamethasone. AB - The effect of dexamethasone on ethanol-induced hypothermia was investigated in 3.5-month old male Wistar rats (N = 10 animals per group). The animals were pretreated with dexamethasone (2.0 mg/kg, i.p.; volume of injection = 1 ml/kg) 15 min before ethanol administration (2.0, 3.0 and 4.0 g/kg, i.p.; 20% w/v) and the colon temperature was monitored with a digital thermometer 30, 60 and 90 min after ethanol administration. Ethanol treatment produced dose-dependent hypothermia throughout the experiment (-1.84 +/- 0.10, -2.79 +/- 0.09 and -3.79 +/- 0.15 degrees C for 2.0, 3.0 and 4.0 g/kg ethanol, respectively, 30 min after ethanol) but only the effects of 2.0 and 3.0 g/kg ethanol were significantly antagonized (-0.57 +/- 0.09 and -1.25 +/- 0.10, respectively, 30 min after ethanol) by pretreatment with dexamethasone (ANOVA, P < 0.05). These results are in agreement with data from the literature on the rapid antagonism by glucocorticoids of other effects of ethanol. The antagonism was obtained after a short period of time, suggesting that the effect of dexamethasone is different from the classical actions of corticosteroids. PMID- 9222407 TI - Distribution of AMPA-type glutamate receptor subunits in the chick visual system. AB - Several glutamate receptor (GluR) subunits have been characterized during the past few years. In the present study, subunit-specific antisera were used to determine the distribution of the AMPA-type glutamate receptor subunits GluR1-4 in retinorecipient areas of the chick brain. Six white leghorn chicks (Gallus gallus, 7-15 days old, unknown sex) were deeply anesthesized and perfused with 4% buffered paraformaldehyde and brain sections were stained using immunoperoxidase techniques. The AMPA-type glutamate receptor subunits GLuR1, GluR2/3 and GluR4 were present in several retinorecipient areas, with varying degrees of colocalization. For example perikarya in layers 2, 3, and 5 of the optic tectum contained GluR1, whereas GluR2/3 subunits appeared mainly in neurons of layer 13. The GluR4 subunit was only detected in a few cells of the tectal layer 13. GluR1 and GluR2/3 were observed in neurons of the nucleus geniculatus lateralis, whereas GluR4 was only present in its neuropil. Somata in the accessory optic nucleus appeared to contain GluR2/3 and GluR4, whereas GluR1 was the dominant subunit in the neuropil of this nucleus. These results suggest that different subpopulations of visual neurons might express different combinations of AMPA type GluR subunits, which in turn might generate different synaptic responses to glutamate derived from retinal ganglion cell axons. PMID- 9222408 TI - Serotonergic neurons in the caudal raphe nuclei discharge in association with activity of masticatory muscles. AB - There is a dense serotonergic projection from nucleus raphe pallidus and nucleus raphe obscurus to the trigeminal motors nucleus and serotonin exerts a strong facilitatory action on the trigeminal motoneurons. Some serotonergic neurons in these caudal raphe nuclei increase their discharged during feeding. The objective of the present study was to investigate the possibility that the activity of these serotonergic neurons is related to activity of masticatory muscles. Cats were implanted with microelectrodes and gross electrodes. Caudal raphe single neuron activity, electrocorticographic activity, and splenius, digastric and masseter electromyographic activities were recorded during active behaviors (feeding and grooming), during quiet waking and during sleep. Seven presumed serotonergic neurons were identified. These neurons showed a long duration action potential (> 2.0 msec) and discharged slowly (2-7 Hz) and very regularly (interspike interval coefficient of variation < 0.3) during quiet waking. The activity of these neurons decreased remarkably during fast wave sleep (78-100%). Six of these neurons showed tonic changes in their activity positively related to digastric and/or masseter muscles activity but not to splenius muscle activity during waking. These data are consistent with the hypothesis that serotonergic neurons in the caudal raphe nuclei play an important role in the control of jaw movements. PMID- 9222409 TI - Inhibition of lipid peroxidation and iron (II)-dependent DNA damage by extracts of Pothomorphe peltata (L). Miq. AB - Leaves of Pothomorphe peltata (L.) Miq. (Piperaceae) are used locally as anti inflammatory, antipyretic, hepatoprotective and diuretic infusions and to treat external ulcers and local infections in several parts of the Peruvian, Bolivian and Brazilian Amazon region. The antioxidant activity of different extracts of P. peltata was studied using the hydroperoxide-initiated chemiluminescence assay in liver homogenates, and the methanolic extract was found to have the highest antioxidant activity, with an IC50 = 4 micrograms/ml. Aqueous and dichloromethane extracts did not show antioxidant activity. The extracts were further evaluated using the thiobarbituric acid-reactive substances (TBARS) assay. Finally, an assay of DNA sugar damage induced by Fe (II) salt was used to determine the capacity of the extracts to suppress the oxidative degradation of DNA. All the extracts showed antioxidant activity in the latter two bioassays. The methanolic extract showed the highest activity in reducing oxidative damage to DNA, with an IC50 = 5 micrograms/ml. Since this extract was highly effective in reducing chemiluminescence and DNA damage, and because the latter activity could be due to the presence of compounds that bind to DNA, DNA-binding activity was studied using the DNA-methyl green (DNA-MG) bioassay. A 30% decrease in the initial absorbance of DNA-MG complex was observed in the methanolic extract at 1000 micrograms/ml, suggesting the presence of compounds that bind to genetic material. No DNA-binding activity was observed in the aqueous or dichloromethane extracts. PMID- 9222410 TI - Role of nitric oxide and superoxide in Giardia lamblia killing. AB - Giardia lamblia trophozoites were incubated for 2 h with activated murine macrophages, nitric oxide (NO) donors or a superoxide anion generator (20 mU/ml xanthine oxidase plus 1 mM xanthine). Activated macrophages were cytotoxic to Giardia trophozoites (approximately 60% dead trophozoites). The effect was inhibited (> 90%) by an NO synthase inhibitor (200 microM) and unaffected by superoxide dismutase (SOD, 300 U/ml). Giardia trophozoites were killed by the NO donors, S-nitroso-acetyl-penicillamine (SNAP) and sodium nitroprusside (SNP) in a dose-dependent manner (LD50 300 and 50 microM, respectively). A dual NO superoxide anion donor, 3-morpholino-sydnonimine hydrochloride (SIN-1), did not have a killing effect in concentrations up to 1 mM. However, when SOD (300 U/ml) was added simultaneously with SIN-1 to Giardia, a significant trophozoite-killing effect was observed (approximately 35% dead trophozoites at 1 mM). The mixtures of SNAP or SNP with superoxide anion, which yields peroxynitrite, abolished the trophozoite killing induced by NO donors. Authentic peroxynitrite only killed trophozoites at very high concentrations (3 mM). These results indicate that NO accounts for Giardia trophozoites killing and this effect is not mediated by peroxynitrite. PMID- 9222411 TI - Cyclosporin inhibits hyperalgesia and edema in arthritic rats: role of the central nervous system. AB - Since arthritis induced by Mycobacterium products (adjuvant) in rats is considered to be immunologically driven, the objective of the present study was to determine if the immunosuppressor drug cyclosporin could affect hindpaw edema and joint hyperalgesia simultaneously. Female Holtzman rats (140-170 g) presented hyperalgesia and edema on the 8th and 12th day following adjuvant injection. Daily systemic (oral or intramuscular) administration of cyclosporin (0.5-5.0 mg kg (-1) day (-1)) or dexamethasone (0.01-0.1 mg kg (-1) day (-1)) for 15 days starting on day zero dose-dependently inhibited the hindpaw edema and hyperalgesia in arthritic rats. However, hyperalgesia but not edema could be detected two days after cyclosporin withdrawal. We concluded that a) the continuous presence of cyclosporin is essential to reduce the development of joint hyperalgesia and that b) different mechanisms underlie the appearance of hyperalgesia and edema in this model. The intracerebroventricular (i.c.v.) administration of 5-50-fold smaller doses of cyclosporin (1.5-150 micrograms/day) or dexamethasone (15 micrograms/day) also reduced the arthritic hindpaw edema and hyperalgesia. Peripheral blood from animals injected with effective systemic cyclosporin doses showed detectable levels of the drug, whereas peripheral blood from those injected with i.c.v. cyclosporin did not, as measured by specific RIA. Our results indicate that cyclosporin administered by the central route is as effective as by the systemic route to reduce joint hyperalgesia and hindpaw edema in arthritic rats. The antiarthritic effect induced by low doses of cyclosporin in the central nervous system (CNS) could be explored to avoid it often associated systemic side effects during chronic therapy. However, the mechanism(s) involved in the antiarthritic response to cyclosporin in the CNS remain to be elucidated. PMID- 9222412 TI - Effects of bromocriptine on serum prolactin levels, pituitary weight and immunoreactive prolactin cells in estradiol-treated ovariectomized rats: an experimental model of estrogen-dependent hyperprolactinemia. AB - The present study was designed to assess the effects of bromocriptine, a dopamine agonist, on pituitary wet weight, number of immunoreactive prolactin cells and serum prolactin concentrations in estradiol-treated rats. Ovariectomized Wistar rats were injected subcutaneously with sunflower oil vehicle or estradiol valerate (50 or 300 micrograms/rat(-1) week (-1) for 2, 4, or 10 weeks. Bromocriptine (0.2 or 0.6 mg rat (-1) day (-1)) was injected daily during the last 5 or 12 days of estrogen treatment. Data were compared with those obtained for intact control rats. Administration of both doses of estrogen increased serum prolactin levels. No difference in the number of prolactin cells in rats treated with 50 micrograms estradiol valerate was observed compared to intact adult animals. In contrast, rats treated with 300 micrograms estradiol valerate showed a significant increase in the number of prolactin cells (P < 0.05). Therefore, the increase inn serum prolactin levels observed in rats treated with 50 micrograms estradiol valerate, in the absence of morphological changes in the pituitary cells, suggests a "functional" estrogen-induced hyperprolactinemia. Bromocriptine decreased prolactin levels in all estrogen-treated rats. The administration of this drug to rats previously treated with 300 micrograms estradiol valerate also resulted in a significant decrease in pituitary weight and number of prolactin cells when compared to the group treated with estradiol alone. The general antiprolactinemic and antiproliferative pituitary effects of bromocriptine treatment reported here validate the experimental model of estrogen induced hyperprolactinemic rats. PMID- 9222413 TI - Baroreflex and chemoreflex dysfunction in streptozotocin-diabetic rats. AB - Several investigators have demonstrated that streptozotocin (STZ) diabetes induces changes in the autonomic control of the cardiovascular system. Changes in cardiovascular function may be related to peripheral neuropathy. The aim of the present study was to analyze changes in heart rate (HR) and arterial pressure (AP) as well as baroreflex and chemoreflex sensitivity in STZ-induced diabetic male Wistar rats (STZ, 50 mg/kg, i.v., 15 days). Intra-arterial blood pressure signals were obtained for control and diabetic rats (N = 9, each group). Data were processed in a data acquisition system (CODAS, 1 kHz). Baroreflex sensitivity was evaluated by measuring heart rate changes induced by arterial pressure variation produced by phenylephrine and sodium nitroprusside injection. Increasing doses of potassium cyanide (KCN) were used to evaluate bradycardic and pressor responses evoked by chemoreflex activation. STZ induced hyperglycemia (447 +/- 49 vs 126 +/- 3 mg/dl), and a reduction in AP (99 +/- 3 vs 118 +/- 2 mmHg), resting HR (296 +/- 11 vs 355 +/- 16 bpm) and plasma insulin levels (16 +/ 1 vs 57 +/- 11 microU/ml). We also observed that the reflex bradycardia (-16.8 +/- 0.1 vs -12.5 +/- 0.1 bpm/mmHg, in the diabetic group) and tachycardia (-3.68 +/- 0.5 vs -1.75 +/- 0.3 bpm/mmHg, in the diabetic group) produced by vasopressor and depressor agents were impaired in the diabetic group. Bradycardia evoked by chemoreflex activation was attenuated in diabetic rats (control: -17 +/- 1, -86 +/- 19, -185 +/- 18, -208 +/- 17 vs diabetic: -7 +/- 1, -23 +/- 5, -95 +/- 13, 140 +/- 13 bpm), as also was the pressor response (control: 6 +/- 1, 30 +/- 7, 54 +/- 4, 59 +/- 5 vs diabetic: 6 +/- 1, 8 +/- 2, 33 +/- 4, 42 +/- 5 mmHg). In conclusion, the cardiovascular response evoked by baroreflex and chemoreflex activation are impaired in diabetic rats. The alterations of cardiovascular responses may be secondary to the autonomic dysfunction of cardiovascular control. PMID- 9222414 TI - Cardiovascular, respiratory and metabolic responses to temperature and hypoxia of the winter frog Rana catesbeiana. AB - The objective of the present study was to determine the effects of hypoxia and temperature on the cardiovascular and respiratory systems and plasma glucose levels of the winter bullfrog Rana catesbeiana. Body temperature was maintained at 10, 15, 25 and 35 degrees C for measurements of breathing frequency, heart rate, arterial blood pressure, metabolic rate, plasma glucose levels, blood gases and acid-base status. Reducing body temperature from 35 to 10 degrees C decreased (P < 0.001) heart rate (bpm) from 64.0 +/- 3.1 (N = 5) to 12.5 +/- 2.5 (N = 6) and blood pressure (mmHg) (P < 0.05) from 41.9 +/- 2.1 (N = 5) to 33.1 +/- 2.1 (N = 6), whereas no significant changes were observed under hypoxia. Hypoxia-induced changes in breathing frequency and acid-base status were proportional to body temperature, being pronounced at 25 degrees C, less so at 15 degrees C, and absent at 10 degrees C. Hypoxia at 35 degrees C was lethal. Under normoxia, plasma glucose concentration (mg/dl) decreased (P < 0.01) from 53.0 +/- 3.4 (N = 6) to 35.9 +/- 1.7 (N = 6) at body temperatures of 35 and 10 degrees C, respectively. Hypoxia had no significant effect on plasma glucose concentration at 10 and 15 degrees C, but at 25 degrees C there was a significant increase under conditions of 3% inspired O2. The arterial PO2 and pH values were similar to those reported in previous studies on non-estivating Rana catesbeiana, but PaCO2 (37.5 +/- 1.9 mmHg, N = 5) was 3-fold higher, indicating increased plasma bicarbonate levels. The estivating bullfrog may be exposed not only to low temperatures but also to hypoxia. These animals show temperature-dependent responses that may be beneficial since during low body temperatures the sensitivity of most physiological systems to hypoxia is reduced. PMID- 9222415 TI - Thyroid function in post-weaning rats whose dams were fed a low-protein diet during suckling. AB - This study was designed to evaluate the thyroid and pituitary hormone levels in post-weaning rats whose dams were fed a low-protein diet during suckling (21 days). The dams and pups were divided into 2 groups: a control group fed a diet containing 22% protein that supplies the necessary amount of protein for the rat and is the usual content of protein in most commercial rat chow, and a diet group fed with a low-protein (8%) diet in which the protein was substituted by an isocaloric amount of starch. After weaning all dams and pups received the 22% protein diet. Two hours before sacrifice of pups aged 21, 30 and 60 days, a tracer dose (0.6 microCi) of 125I was injected (i.p.) into each animal. Blood and thyroid glands of pups were collected for the determination of serum T4, T3 and TSH and radioiodine uptake. Low protein diet caused a slight decrease in radioiodine uptake at 21 days, and a significant decrease in T3 levels (128 +/- 14 vs 74 +/- 9 ng/dl, P < 0.05), while T4 levels did not change and TSH was increased slightly. At 30 days, T3 and TSH did not change while there was a significant increase in both T4 levels (4.8 +/- 0.3 vs 6.1 +/- 0.2 micrograms/dl, P < 0.05) and in radioiodine uptake levels (0.34 +/- 0.02 vs 0.50 +/- 0.03%/mg thyroid, P < 0.05). At 60 days serum T3, T4 and TSH levels were normal, but radioiodine uptake was still significantly increased (0.33 +/- 0.02 vs 0.41 +/- 0.03%/mg thyroid, P < 0.05). Thus, it seems that protein malnutrition of the dams during suckling causes hypothyroidism in the pups at 21 days that has a compensatory mechanism increasing thyroid function after refeeding with a 22% protein diet. The radioiodine uptake still remained altered at 60 days, when all the hormonal serum levels returned to the normal values, suggesting a permanent change in the thyroid function. PMID- 9222416 TI - Partial characterization of folate uptake in microvillous membrane vesicles isolated from human placenta. AB - The purpose of the present study was to determine biochemical parameters of folate uptake, and the putative contribution of the membrane-anchored folate receptor in microvillous membrane vesicles obtained from the syncytiotrophoblast of human term placenta. Uptake of [3H]-pteroylglutamic acid (PGA) by microvillous membrane vesicles was pH dependent with a maximum at pH 6.0, and attained equilibrium at 60 min of incubation. Uptake was higher in the presence on an inward pH gradient (pHout = 6.0; pHin = 7.5) than in the absence of the gradient (pHout = pHin = 6.0). The effect of changes in medium osmolality showed that both binding to the vesicular membrane and internalization contributed to the measured [3H]-PGA uptake. Equilibrium uptake experiments using [3H]-PGA concentrations within the physiological range of folate in blood serum showed that saturation was achieved at 30 nM and revealed a single class of binding sites with a Kd of 1.8 nM for [3H]-PGA. Cleavage of the glycosyl-phosphatidylinositol moiety of the folate receptor, which anchors the receptor to the membrane, with phosphatidylinositol-specific phospholipase C resulted in a reduction of about 80% in [3H]-PGA uptake. In conclusion, our results showed that the folate uptake in the maternally facing membrane of the human placenta presents a saturable component and is mediated by the folate receptor to ensure an adequate maternal fetal folate transfer. PMID- 9222417 TI - Insulin receptor tyrosine kinase activity in colon carcinoma. AB - Colon carcinoma is the most common tumor of the gastrointestinal tract. According to some investigators, insulin, epidermal growth factor (EGF) and insulin-like growth factor I (IGF-I) man be involved in the neoplastic proliferation. Insulin binding and receptor tyrosine kinase activity were investigated in colon carcinomas and in normal colons. The insulin receptor concentration, as shown by binding assays, was 17.4 +/- 4.3 fmol/micrograms in normal colon and 29.69 +/- 9.4 fmol/micrograms in colon carcinoma. Nevertheless, the insulin affinity of the receptor was similar in both groups (Kd identical to 1 nM). Both normal and neoplastic colon showed phosphorylation of the insulin receptor. The electrophoretic migration of the beta-subunit of the insulin receptors purified from colon carcinomas was similar to that of normal colon and both tissues demonstrated an insulin-dependent autophosphorylation. The receptor tyrosine kinase activity was measured by the incorporation of [gamma 32P]ATP into the beta subunit. The basal and the insulin-stimulated tyrosine kinase activities were significantly higher in colon carcinomas compared to normal colon tissues (2.2 and 1.6 times, respectively). Understanding the metabolism of neoplastic cells may contribute to the development of prevention strategies as well as new therapies. It is now necessary to study other steps of the insulin signal transduction pathway, such as insulin receptor substrate 1 phosphorylation. PMID- 9222419 TI - Different urinary albumin responses to submaximal exercise by normoalbuminuric diabetic children and controls. AB - It is not clear if exercise could be useful to identify diabetic patients at risk for the development of nephropathy. We evaluated the responses of blood pressure (BP) and urinary albumin (Alb) and retinol-binding protein (RBP) excretion to standardized sub-maximal exercise in 17 normoalbuminuric normotensive children with IDDM and 17 matched normal subjects. RBP was used as an index of tubular function. Standardization of exercise load was based on heart rate (HR) which was maintained at 70% of the maximum calculated to age. A step exercise test lasted for 35 min; baseline BP and HR were taken at midtime and during cooling down. Pre and postexercise urines were obtained for Alb, RBP and creatinine determinations. Both groups showed a significantly increased systolic BP at the midpoint but the percent variations were not different. HR responses did not differ and demonstrated the exercise effectiveness. Great variability in Alb excretion was observed within the normal range for both groups. The baseline Alb/creatine ration was not significantly different between normal and diabetic subjects, but became different following exercise (6.6 +/- 4.1 vs 17.7 +/- 18.7 mg/g. P < 0.05). While this ratio decreased in the control group (14.8 +/- 11.1 to 6.6 +/- mg/g, P < 0.02), it increased (9.0 +/- 7.1 to 17.7 +/- 18.7 mg/g, P = 0.05) in diabetic patients. Percent variations in the two groups occurred in opposite directions and were significantly different. RBP/creatinine followed the same pattern within each group; normals showed a tendency to a decrease (0.058 +/ 0.064 to 0.030 +/- 0.039 microgram/g, P = 0.05) and diabetic patients to an increase (0.116 +/- 0.125 to 0.247 +/- 0.247 microgram/g, P = 0.06). We conclude that there was a variable proteinuric response to exercise among diabetic subjects with normal renal function as evaluated by albumin excretion. A subset of IDDM patients responded abnormally to the exercise stress, increasing albumin excretion to levels compatible with microalbuminuria. Whether this heterogeneity reflects individual risk for diabetic renal disease require further investigation. PMID- 9222418 TI - Cloning and sequencing of the nitrogenase structural genes nifHDK of Herbaspirillum seropedicae. AB - The nitrogenase structural genes (nifHDK) of the endophytic diazotroph Herbaspirillum seropedicae were isolated from a genomic bank by plate hybridization. Sequence analysis of the DNA showed a consensus promoter region upstream for the nifH gene containing a -24/-12 type promoter together with NifA- and integration host factor (IHF)- binding sites. The derived protein sequences of NifH, NifD and NifK contained conserved cysteine residues for binding iron sulfur clusters and the iron-molybdenum cofactor. These protein sequences showed the strongest similarities to the nifHDK gene products of the symbiotic diazotroph Bradyrhizobium japonicum (93.5%, 91.3% and 83.3%, respectively), the plant-associated diazotroph Azospirillum brasilense (90.0%, 83.7% and 75.1%, respectively) and to Thiobacillus ferrooxidans (91.0%, 83.4% and 81.1%, respectively) of the same phylogenetic group of the protobacteria. PMID- 9222420 TI - Effect of fasting on insulin signaling in the aorta of intact rats. AB - Insulin stimulates the tyrosine kinase activity of its receptor, resulting in the phosphorylation of its cytosolic substrate, insulin receptor substrate 1 (IRS-1). Previous studies have demonstrated a tissue-specific regulation of IRS-1. In the present study we investigated the levels and phosphorylation state of IRS-1 after insulin stimulation in the rat aorta in vivo, and the modulation of this protein after 72 h of fasting, using immunoprecipitation and immunoblotting with anti insulin receptor, anti-IRS-1 and antiphosphotyrosine antibodies. We show that IRS 1 is present in rat aorta, and is tyrosine phosphorylated after insulin stimulation. After insulin stimulation, rats fasted for 72 h showed an increase in insulin receptor (100 +/- 45%, P < 0.05) and IRS-1 phosphorylation (68 +/- 24%, P < 0.05) in aorta, compared to fed rats. There was no change in insulin receptor of IRS-1 protein levels in fasted rats. In summary, the present study demonstrated that proteins involved in the early steps of insulin signal transduction are present in the rat aorta and can be modulated by fasting. It will be of interest to study the regulation of these proteins in the aorta of animal models of hypertension and/or atherosclerosis. PMID- 9222421 TI - Prevalence of anemia among school-children from Rio Acima (State of Minas Gerais, Brazil): use of the standardized prevalence method and evaluation of iron deficiency. AB - The prevalence of anemia and iron deficiency was investigated in 332 children aged 7 to 15 years, 156 (47%) boys and 176 (53%) girls enrolled in the schools of the municipality of Rio Acima, MG. Seventy-four children were white (22.3%), 218 were mulatto (65.7%), and 40 were black (12%). Mean hemoglobin level was 12.75 +/ 0.75 g/dl. Lower values were determined for black children (12.32 +/- 0.87g/dl) compared to white (12.76 +/- 0.99 g/dl) and mulatto (12.81 +/- 0.94 g/dl) children. The prevalence of anemia was 16.6% when determined on the basis of the percentage of children with hemoglobin values lower than the 3rd percentile for age and sex (standard method), and 36.2% when determined by the standardized prevalence method for the evaluation of the prevalence of malnutrition in populations. Depletion of iron reserves was 8.13% for the population in general and 20% for the anemic children. This low prevalence of iron deficiency may have been the result of the value adopted as the lower normal limit (10 ng/ml) for serum ferritin values. The small percentage of anemic children with iron depletion may also be justified by the standard of normality adopted for hemoglobin values which was originally elaborated for the white population of North America and Finland and therefore may be inadequate for the population studied here, of diverse racial composition. PMID- 9222422 TI - Monoclonal antibodies against low density lipoprotein with various degrees of oxidative modifications. AB - Oxidative processes leading to the generation of oxidized low density lipoprotein (oxLDL) particles have been suggested to be an important factor in the pathogenesis of atherosclerosis. After initiation of the oxidative process, LDL undergoes a progressive protein and lipid fragmentation. To understand this process and the role of oxLDL in various diseases of inflammatory origin, we have generated mouse monoclonal antibodies against copper-oxidized human LDL. Mice were immunized intrasplenically and after one intravenous boost the spleen cells were fused with the Sp2/0 hybridoma fusion partner. The hybridoma clones obtained after selection and cloning were analyzed for reactivity against oxLDL with various degrees of copper-mediated oxidative modifications. Three hybridoma clones were purified and further characterized. The following observations were made: 1) the intrasplenic route of immunization, avoiding the use of mycobacterial adjuvants, yielded a high frequency of positive clones; 2) the individual hybridomas reacted against LDL with various degrees of oxidative modifications; 3) the monoclonal antibodies could be used in ELISA and to detect oxLDL in immunohistochemical tissue staining, and 4) the monoclonal antibodies also detected oxLDL from hamsters and rabbits. We conclude that these monoclonal antibodies could be useful to further investigate the role of oxLDL in inflammation and in the immune response. PMID- 9222423 TI - Hepatitis C virus genotypes in Southern Brazil. AB - The prevalence of hepatitis C virus (HCV) genotypes in Southern Brazil was studied in the plasma of 100 HCV-RNA-positive patients attended in Porto Alegre, South of Brazil. Reverse transcription-polymerase chain reaction (RT-PCR) products from the 5' noncoding region were double digested with RsaI-HaeIII and BstNI-HinfI and analyzed by restriction fragment length polymorphism (RFLP). Three genotypes (1, 2 and 3) were demonstrable, the most prevalent being HCV type 1 (55 of 100 patients, 55%), followed by HCV type 3 (37 of 100 patients, 37%) and HCV type 2 (8 of 100 patients, 8%). There was an unusual high prevalence of genotype 3, in contrast to the majority of published data from the Southeast region. PMID- 9222424 TI - Complement resistance of Leishmania amazonensis promastigotes is independent of parasite proteases and lysis of sensitive forms is not due to natural antibodies in normal human serum. AB - Leishmania amazonensis promastigotes cultivated in vitro differentiate from complement-sensitive to complement-resistant forms. In order to determine the possible involvement of parasite proteases in this process, L. amazonensis promastigotes were collected daily and their proteolytic enzyme patterns analyzed using polacrylamide gels copolymerized with gelatin. Although promastigotes at different growth stages showed differences in protease patterns, these changes did not correlate with their susceptibility to complement. The major protease of promastigotes, gp63, was expressed at the same level throughout culture, regardless of the complement resistance of the promastigotes. Furthermore, inhibitors specific for the classes of proteases found in L. amazonensis promastigotes did not interfere with the complement-mediated killing of promastigotes. We also investigated the binding of natural antibodies to promastigotes. at different stages of growth using ELISA. Although complement sensitive promastigotes bound significantly more antibodies from fresh normal human serum than complement-resistant promastigotes, equivalent amounts of C3 were detected on their surfaces following complement activation. Moreover, serum depleted of anti-Leishmania antibodies was an efficient in killing promastigotes and the intact serum. These data suggest that the resistance of L. amazonensis to complement killing involves strategies other than that of the regulated expression of endogenous proteases capable of inactivating complement components, or the differential ability to bind natural antibodies that might interfere with complement deposition on the parasite surface. PMID- 9222426 TI - Correlation between concomitant theta waves in nucleus reticularis pontis oralis and in hippocampus, thalamus and neocortex during dreaming in rats. AB - Theta waves, which are the main electrophysiological expression of dreaming activity in many brain structures of rats, often undergo specific changes in voltage and frequency according to the oniric patterns. Much is known about their mechanisms but little is known regarding their origin, which has been ascribed to a specific activation of either the reticular formation or the septal nuclei or nucleus reticularis pontis oralis. In the present study, rats were prepared for chronic recording of the electro-oscillograms of cortical areas 10, 3 and 17, of hippocampal CA1 and CA3 fields, of nucleus reticularis thalami, nucleus reticularis pontis oralis and occasionally of nucleus reticularis caudalis. Head, rostrum, eye and forelimb movements were also recorded, so that the oniric behaviors could be precisely identified. The scatter diagrams and the corresponding correlation coefficients (r) of the voltage of concomitant waves were determined for each possible pair of leads. The potentials were analyzed at a frequency of 256 Hz over a period of 1 to 3 sec. A very high degree of correlation was observed between theta waves in nucleus reticularis pontis oralis, hippocampal fields and nucleus reticularis pontis caudalis; sometimes r approached unity. Although these data cannot be taken as proof of nucleus reticularis pontis oralis being the source of theta waves, they are at least compatible with this hypothesis. PMID- 9222425 TI - Preparation of a heterologous antiserum for the determination of von Willebrand factor in human plasma. AB - A simple method for the preparation of rabbit antiserum against human von Willebrand factor (vWF) from commercial lyophilized factor VIII concentrate is described. vWF antigen (vWFAg)-like protein was obtained by gel filtration of the concentrate on Sepharose 4B. A combination of measurements of protein content by absorbance at 280 nm, and of vWFAg by electroimmunoassay using a commercial antibody, provided the data needed to select the Sepharose-filtered fractions with the highest concentrations of vWFAg-like protein. The immunization scheme used induced high antibody titers from the 45th to the 126th day after the first immunization. The resulting antiserum showed a performance similar to that of a commercial preparation in terms of vWFAg determination by electroimmunoassay and two-dimensional crossed-immunoelectrophoresis. PMID- 9222427 TI - Calcium channel blockers inhibit the antidipsogenic effect of central injections of zinc in rats. AB - Previous data from our laboratory have indicated that acute third ventricle injections of Zn2+ elicit a significant antidipsogenic response in rats in three different situations; dehydration, and central angiotensinergic or cholineric stimulation. In the present study we analyzed whether this response depends on voltage-dependent calcium channels. Dehydrated (14 h of water deprivation, overnight) animals received 2-microliters i.c.v. injections of zinc acetate (Zn(Ac)2; 300 pmol/rat) after pretreatment with the voltage-dependent calcium channel blockers gadolinium (Gd3+; 0.03, 3.0 and 30 pmol/rat) or verapamil (VER; 0.027, 0.05 and 0.11 pmol/rat). Both blockers reserved the antidipsogenic effect of third ventricle injections of Zn2+ in a dose-dependent manner. After 120 min, animals pretreated with saline receiving Zn(Ac)2 drank 3.10 +/- 0.57 ml/100 g body weight while those pretreated with Gd3+ at the highest dose displayed a water intake of 5.45 +/- 0.41 ml/100 body weight (P < 0.01). Animals pretreated with the vehicle of VER receiving Zn(Ac)2 drank 3.15 +/- 0.45 ml/100 g while animals pretreated with VER at the highest dose receiving Zn(Ac)2 drank 6.16 +/- 0.62 ml/100 g (P < 0.01). The antidipsogenic effect of Zn(Ac)2 seems to be specific since the metal (same dose and injection procedures) did not modify food intake in rats after 24 h of food deprivation. It is suggested that Zn2+ exerts its antidipsogenic effect by activation of mechanism(s) depending on the functional integrity of voltage-dependent calcium channels. PMID- 9222428 TI - Territorial aggression, body weight, carbohydrate metabolism and testosterone levels of wild rats maintained in laboratory colonies. AB - Aggressive territorial behavior was studied in 15 colonies of wild rats (Rattus norvegicus), each consisting of 2 males and 1 female. One of the males attacked an intruder rat more frequently and had a higher body weight than the less aggressive one. In another experiment, male and female rats were raised in colonies from weaning to adulthood. Animals were weighed every 7 days until 90 days of age when plasma testosterone was measured in males, and plasma glucose, hepatic and muscle glycogen were measured in both males and females. THe heavier (and thus possibly dominant) males in the colonies of 3 males and 1 female also had a higher body weight than males raised with females, but without any male partner. In this long-term social relationship there were no significant differences in carbohydrate metabolism among the animals. The differential growth rate among males was established around the period of sexual maturity. Moreover, when adult, heavier males had higher plasma testosterone levels compared to the other members of the colony and also to males that had no other competitive male partner. This higher androgenic hormone level may be one of the causal factors involved in the weight increase of the dominant male in the colony. PMID- 9222429 TI - Antagonism of clonidine injected intracerebroventricularly in different models of salt intake. AB - Clonidine, an alpha 2-adrenergic agonist, injected into the brain inhibits salt intake of animals treated by the diuretic model of sodium depletion. In th present study, we address the question of whether central injection of clonidine also inhibits salt intake in animals deprived of water or in the need-free state. Saline or clonidine (30 nmol) was injected into the anterior third ventricle of 24-h sodium-depleted (furosemide + removal of ambient sodium), of 24-h water deprived and of normovolemic (need-free state) adult male rats. Clonidine injected intracerebroventricularly (i.c.v.) inhibited the 1.5% NaCl intake for 1209 min by 50 to 90% in every model tested. Therefore, different models of salt intake are inhibited by i.c.v. injection of clonidine. Idazoxan, an alpha 2 adrenergic antagonist, injected i.c.v. at a dose of 160 nmol, inhibited the effect of clonidine only in the furosemide + removal of ambient sodium model of salt intake. This indicates that the antagonism of this effect by idazoxan is dependent on the body fluid/sodium status of the animal. PMID- 9222430 TI - Sodium balance of hyper- and hypo-natriophilic rats under basal conditions and during sodium deprivation. AB - Sodium and water balance was determined in two strains of Wistar rats selectively bred for high (hypernatriophilic, HR) or low salt preference (hyponatriophilic, HO) under basal conditions and during sodium deprivation. Male rats from each stain were selected for an average ingestion of 1.5% NaCl solution of more than (HR) or less than (HO) 4 ml 100 g body weight (-1) day (-1), during a 10-day period. HR rats (N = 17) presented markedly higher sodium intake under basal conditions (2.983 +/- 0.316 mEq 100 g body weight (-1) day (-1)) than HO rats (N = 12; 0.406 +/- 0.076 mEq 100 g body weight (-1) day (-1); Mann-Whitney test, P < 0.01). Water (HR: 8.6 +/- 0.57; HO: 7.7 +/- 0.32 ml 100 g body weight (-1) day ( 1)) and sodium balances (HR: 0.936 +/- 0.153; HO: 0.873 +/- 0.078 mEq 100 g body weight (-1) day (-1)) were similar in both strains, despite a higher sodium and total fluid (HR: 16.3 +/- 1.06; HO: 10.8 +/- 0.49 ml 100 g body weight (-1) day ( 1); P < 0.01) ingestion in HR rats. During sodium deprivation HR rats (N = 13) exhibited a sodium balance similar to that of HO rats (N = 13) (HR: -0.159 +/- 0.011; HO: -0.129 +/- 0.019 mEq 100 g body weight (-1) day (-1)), and, in addition, an adequate suppression of natriuresis (HR: 0.049 +/- 0.011; HO: 0.026 +/- 0.004 mEq 100 g body weight (-1) day (-1)). These data show that HR rats present hypernatriophilia as a primary trait, since their sodium-conserving mechanisms are intact. Therefore, these rats provide an adequate model to study factors that determine innate sodium preference. PMID- 9222431 TI - Central interaction between atrial natriuretic peptide and angiotensin II in the control of sodium intake and excretion in female rats. AB - We investigated the effects of estrogen on sodium intake and excretion induced by angiotensin II (ANG II), atrial natriuretic peptide (ANP) or ANG II plus ANP injected into the median nucleus (MnPO). Female Holtzman rats weighing 250-300 g were used. Sodium ingestion and excretion 120 min after the injection of 0.5 microliters of 0.15 M NaCl into the MnPO were 0.3 +/- 0.1 ml (N = 12) and 29 +/- 7 microEq in intact rats, 0.5 +/- 0.2 ml (N = 10) and 27 +/- 6 microEq in ovariectomized rats, and 0.2 +/- 0.08 (N = 11) and 36 +/- 8 microEq in estrogen treated ovariectomized (50 micrograms/day for 21 days) rats, respectively. ANG II (21 microM) injection in intact, ovariectomized, and estrogen-treated ovariectomized rats increased sodium intake (3.8 +/- 0.4, 1.8 +/- 0.3 and 1.2 +/- 0.2 ml/120 min, respectively) (N = 11) and increased sodium excretion (166 +/- 18, 82 +/- 22 and 86 +/- 12 microEq/120 min, respectively) (N = 11). ANP (65 microM) injection in intact (N = 11), ovariectomized (N = 10) and estrogen treated ovariectomized (N = 10) rats increased sodium intake (1.4 +/- 0.2, 1.8 +/ 0.3, and 1.7 +/- 0.3 ml/120 min, respectively) and sodium excretion (178 +/- 19, 187 +/- 9, and 232 +/- 29 microEq/120 min, respectively). Concomitant injection of ANG II and ANP into the MnPO of intact (N = 12), ovariectomized (N = 10) and estrogen-treated ovariectomized (N = 10) rats caused smaller effects than those produced by each peptide given alone: 1.3 +/- 0.2, 0.9 +/- 0.2 and 0.3 +/- 0.1 ml/120 min for sodium intake, respectively, and 86 +/- 9, 58 +/- 7, and 22 +/- 4 microEq/120 min for sodium excretion, respectively. Taken together, these results demonstrate that there is an antagonistic interaction of ANP and ANG II on sodium intake and excretion, and that reproductive hormones affect this interaction. PMID- 9222432 TI - Reliability of the Portuguese version of the structured clinical interview for DSM-III-R (SCID) in a Brazilian sample of psychiatric outpatients. AB - The objective of the present study was to determine the reliability of psychiatric diagnoses using a translation and adaptation of Portuguese of the "Structured Clinical Interview for DSM-III-R-patient version" (SCID-P) and the "Structured Clinical Interview for DSM-III-R Personality Disorders" (SCID-II), using the joint interviews methodology. Thirty-nine subjects were evaluated using the SCID-P and 20 of them using the SCID-II. Interrater reliability was analyzed statistically by means of the Kappa Coefficient. Agreement between results obtained with SCID-P was statistically significant for the major diagnostic categories of DSM-III-R and for 10 of the 12 specific diagnostic categories studied (a minimum of 4 subjects per diagnosis). Agreement was not statistically significant for Psychotic Disorder Not Otherwise Specified (NOS) and for Other Bipolar Disorder. The Weighted Kappa for the main diagnoses and the Overall Kappa for the entire set of 25 specific diagnostic categories proposed by the SCID-P were statistically significant. The general agreement for Personality Disorders with SCID-II was statistically significant. The Kappa Coefficient was determined for the Avoidant, Paranoid, Histrionic and Borderline Personality Disorders and for the Conduct Disorder. The remaining Personality Disorders were not analyzed statistically because of their low prevalence in the sample. Agreement was not significant only for the Histrionic Personality Disorder. These data suggest that the translation and adaptation of the SCID-P and SCID-II to Portuguese presents, in general, good reliability indices, and thus its use is recommended. PMID- 9222433 TI - Contractility changes of the right and ventricular muscle after chronic myocardial infarction. AB - Contractility changes and adaptive responses resulting from acute left ventricular (LV) myocardial infarction are not restricted to the LV myocardium. The reduced LV function increased the right ventricular (RV) pressure load and neurohumoral factors, activated by the infarction PMID- 9222434 TI - Diameter of the mammalian porin channel in open and "closed" states: direct measurement at the single channel level in planar lipid bilayer. AB - Porin isolated from bovine skeletal muscle was reconstitute in planar lipid bilayers under voltage clamp conditions. A set of non-electrolytes were used as molecular probes for determining the pore diameter. The maximal diameter of the open channel was estimated to be 3.02 +/- 0.26 nm. As observed for other porin channels, a large transmembrane potential drove the channel into a "closed" state. The channel transition to the low conductance (closed) state was followed by a decrease in the maximal diameter of the channel to 2.4 +/- 0.08 nm. PMID- 9222435 TI - A method to estimate mitochondrial Ca2+ uptake in intact cardiac myocytes. AB - In the present paper we describe a method to estimate mitochondrial Ca2+ uptake during the declining phase of Ca2+ transients (cell relaxation) in intact isolated myocardial cells. This method is based on inhibition of sarcoplasmic reticulum (SR) Ca2+ accumulation by caffeine, blockade of Ca2+ transport via sarcolemmal Ca(2+)-ATPase by treatment with carboxyeosin and inhibition of sarcolemmal Na+/Ca2+ exchange by removal of extracellular Na+ and Ca2+.Ca2+ transients were evoked in rabbit ventricular myocytes by quick and sustained caffeine application (10 mM) after a 5-min period of electrical stimulation to load the SR with Ca2+. Mitochondrial Ca2+ transport was estimated using a model described by Sipido and Wier (Journal of Physiology (1991), 435: 605-630), which was originally proposed to describe Ca2+ fluxes during excitation-contraction coupling in cardiac cells. Our results indicate that, in intact rabbit myocytes, the Ca2+ flux due to net mitochondrial Ca2+ uptake may attain a value close to 1 microM/sec. PMID- 9222436 TI - Possible modulatory role of non-activated lymphocytes on insulin secretion. AB - In order to study the probable physiological role of non-activated lymphocytes on islet B-cells, we incubated and perfused rat pancreatic islets in the presence of low (2.8 mM) and high (16.7 mM) glucose concentrations after pre-exposure for 60 min to rat lymphocytes or to substances secreted by lymphocytes. Insulin secretion and 86Rb+, 45Ca2+ and [3H]-phosphoinositide metabolite fluxes were lower compared to controls when islets were pre-exposed to lymphocytes but were not different when islets were pre-exposed to substances secreted by lymphocytes. These alterations in isotope flux suggest that, when lymphocytes and islets are in contact, closure of potassium channels and a paradoxical effect of glucose load on insulin release occur in the presence of low glucose concentrations. The alterations observed are probably due to a swift and direct action of lymphocyte secretion perhaps induced by a direct of islet cells. PMID- 9222437 TI - Oxidative stress: animal adaptations in nature. AB - As a consequence of aerobic life, an organism must deal with the continuous generation of reactive oxygen species (O2-, H202, .OH) as byproducts of metabolism and defend itself against the harm that these can do to cellular macromolecules. Organisms protect themselves from such damage with both enzymatic and nonenzymatic antioxidant defenses. However, the reperfusion injuries noted after ischemic insult in mammalian organs and ascribed to a burst of reactive oxygen species produced when oxygenated blood is reintroduced demonstrate that the antioxidant defenses of many organisms can be overwhelmed, Although unusual among most mammals, many organisms routinely experience wide variation in oxygen availability to their tissues due to factors such as environmental oxygen lack, breath-hold diving, extracellular freezing, or apnoeic breathing patterns in arrested metabolic states. In recent studies using various animal models (anoxia tolerant turtles, freeze-tolerant snakes and frogs, estivating snails) our laboratory has explored the adaptations of antioxidant defenses that allow such organisms to deal with rapid changes in tissue oxygenation with little or no accumulation of damage products. The key to successful transitions in several systems is the induction, during the oxygen-limited state, of elevated activities of antioxidant and associated enzymes, such as catalase, superoxide dismutase, glutathione-S-transferase, and glutathione peroxidase, so that damage during the reintroduction of oxygen (such as lipid peroxidation) is minimized. However, animals that are excellent facultative anaerobes, such as freshwater turtles, appear to deal with potential of oxidative stress during the anoxic-aerobic transition by maintaining constitutively high antioxidant defenses (e.g. enzyme activities similar to those of mammals and much higher than those of anoxia intolerant lower vertebrates) that can readily accommodate the burst of reactive oxygen species generation when breathing is renewed. PMID- 9222438 TI - Fish antioxidant defenses--a comparative approach. AB - There are few comparative studies of vertebrate antioxidant defenses (AD) in the literature. Enzymatic (superoxide dismutase, SOD, and catalase, CAT) and non enzymatic (alpha-tocopherol, beta-carotene, ubiquinol 10 and blood glutathione) antioxidant defenses were investigated in the liver and blood of 37 fish species, 15 marine species of the southeastern Brazilian coast and 22 freshwater species from the Central Amazon basin. More active marine species displayed in general higher concentrations of SOD and CAT in the liver and blood, compared to those of sedentary or bottom-dwelling species. AD status in marine fish m ay be related to the oxygen consumption of the tissues and of the whole organism, while in freshwater AD may be related to physical and chemical characteristics of the environment rather than to activity level. As thermoconformer organisms, most fish must routinely cope with environmental temperature changes and, consequently, with changes in their metabolic rates. The relatively high antioxidant defense levels that typify fishes, even when compared to endotherms such as birds and mammals, may protect aquatic organisms against the consequences of temperature oscillations. PMID- 9222439 TI - Cellular signalling in vertebrate pigment cells. AB - Chromatophores are specialized integumental stellate cells that synthesize and store pigments. Pigment granules are translocated within chromatophores of poikilothermic vertebrates and crustaceans in response to photic, thermal and/or neurohormonal stimuli, allowing the animal to rapidly change color for thermoregulation, adaptation to light and background, and social behavior display. Birds and mammals do not show color changes, but may present slow long term responses, such as melanocyte proliferation, melanin synthesis and melanin granule translocation into feathers, hair and surrounding keratinocytes. Pigment translocation in lower vertebrates as well as pigment production in all vertebrates are modulated by a variety of hormones and neurotransmitters acting on transmembrane receptors located on the cell surface. Alpha-melanocyte stimulating hormone (alpha-MSH), melanin-concentrating hormone (MCHA), melatonin and catecholamines are the most important pigment cell agonist in vertebrates. The major signalling pathway leading to pigment dispersion and melanin synthesis appears to be involve stimulation of adenylate cyclase followed by an increase in the cAMP level and activation of cAMP-dependent protein kinases (PKAs). Another melanogenesis-related intracellular pathway involves the activation of protein kinase C (PKC) by diacylglycerol, and the increase in cytosolic Ca2+ by inositol triphosphate. Growth factors such as basic fibroblast growth factor (bFGF), hepatocyte growth factor (HGF) and mast cell growth factor (MGF or KIT ligand), and UV radiation modulate the melanogenic and mitogenic processes in vertebrate melanocytes as well. PMID- 9222440 TI - Use of cDNA cloning to study the mechanism of action of glucocorticoid hormones at the molecular level. AB - We have previously described a dramatic phenomenon of phenotypic reversion (tumor to normal) caused by glucocorticoid hormones in C6/ST1 rat glioma cells, but not in their hormone-resistant C6/P7 counterpart. Blind cDNA cloning was adopted to search for glucocorticoid-regulated gene sequences responsible for this phenotype reversion. Differential hybridization and differential display of RNA were used in parallel to isolate a number of cDNA clones that were characterized by DNA sequencing and Northern blot analysis. This approach was coupled to the analysis of known growth control genes (oncogenes, tumor suppressor genes, cyclins, cyclin dependent kinases, other kinases). Glucocorticoid target genes isolated from this cell system are likely to be good anti-tumor candidate molecules which can be used in tumor therapy and anti-tumor drug design. PMID- 9222441 TI - Use of neurotoxins to study Ca2+ channel functions. AB - The article contains a brief review on the properties and classification of voltage-dependent Ca2+ channels and on the organic blockers of the different channel types. The effects of peptide toxins from the venoms of Conus sp and of the spider Agelenopsis aperta on high voltage-activated Ca2+ channels are discussed. In addition, we present preliminary data on a novel peptide toxin purified from the venom of the spider Phoneutria nigriventer, which is a powerful blocker of L- and N-type Ca2+ channels. PMID- 9222442 TI - Toxins affecting K+ channels. AB - Potassium channels are involved in modulating the excitability of neurones by regulating the membrane potential, or by affecting the amount of neurotransmitter released from nerve terminals. Potassium channels are highly diverse and can be activated by either voltage or increased intracellular calcium concentration. The potassium channel forms a highly selective membrane pore. Four subunits each with six membrane-spanning regions (S1-S6) are required to produce a functional pore. Molecular biologists have cloned more than 50 different potassium channel subtypes. Naturally occurring protein toxins have been used to pharmacologically characterize native and cloned potassium channels. PMID- 9222443 TI - 5 alpha reductase activity in human gingiva and gingival fibroblasts in response to bacterial culture supernatants, using [14C]4-androstenedione as substrate. AB - 5 alpha-Reduction of androgen substrates is increased in inflamed gingiva. It was therefore relevant to study the effect of bacterial culture supernatants derived from Prevotella intermedius (P.i), Porphyromonas gingivalis (P.g) and Actinobacillus actinomycetemcomitans (A.a) on the metabolism of [14C]4 androstenedione to 5 alpha-dihydrotestosterone (DHT) in gingival tissue and cultured fibroblasts. Chronically inflamed human gingival tissue and cultured gingival fibroblasts from the same source were incubated in duplicate with [14C]4 androstenedione and optimal concentrations of P.i, P.g. and A.a culture supernatants in Eagle's minimal essential medium in a CO2 incubator for 24 h at 37 degrees C. The metabolites were then extracted, separated and quantified using a radioisotope scanner. There were 87, 50 and 6% increases in DHT synthesis by human gingival tissue in response to the culture supernatants of P.i, P.g and A.a, respectively, over control incubations (n = 3; p < 0.01: Wilcoxon signed ranked statistic for paired observations). With the cells in culture, all four fibroblast cell lines produced DHT and testosterone from [14C]4-androstenedione in varying amounts. The production of DHT was enhanced in the presence of each the bacterial culture supernatants to varying degrees (P.i 40%, P.g 35% and A.a 40%; p < 0.01). Combinations of the bacterial extracts: (P.i + P.g), (P.i + A.a), (P.g + A.a) and (P.i + P.g + A.a) showed intermediate or suppressor effects on DHT formed compared with individual incubations. Culture supernatants of these pathogens can influence DHT synthesis in fibroblasts, and effect that is modulated by baseline androgen metabolism and the proportion of virulent pathogens present. This may have some bearing on host susceptibility on host and the progression of the periodontal lesion. PMID- 9222444 TI - Stimulation of plasminogen activator/plasmin system in gingival fibroblast cells by oxygen radicals. AB - The effect of extracellular oxygen radicals on cultured gingival fibroblast cells (Gin cells) was investigated in the plasminogen activator (PA)/plasmin system. The activation of the PA/plasmin system in Gin cells exposed to a sublethal oxygen radical [hypoxanthine (HX) 0.1 mg ml-1/xanthine oxidase (XOD) 5 munit ml 1] system was examined. Following a 1 h exposure, washed cells were cultured for up to 24 h in fresh medium containing 2% fetal calf serum. The exogenous addition of superoxide dismutase, an oxygen radical scavenger, abolished the PA/plasmin activity enhanced by the HX/XOD system. The PA produced by Gin cells was found to be urokinase-type PA (uPA), as preincubation of Gin cell-conditioned medium with anti-uPa serum completely inhibited PA activity. These findings suggest that extracellular oxidant targetting to Gin cells may be involved in the progression of inflammation and the invasion of periodontium through stimulation of the PA/plasmin system. PMID- 9222445 TI - Enhancement by sodium orthovanadate of the formation and mineralization of bone nodules by chick osteoblasts in vitro. AB - Orthovanadate is a known inhibitor of phosphotyrosyl protein phosphatase and is reported to stimulate osteogenic cell proliferation and differentiation when administered during the logarithmic growth phase and to potentiate the mitogenic effects of several growth factors. There is little information concerning the effects of orthovanadate on bone matrix deposition and mineralization, although there is some evidence that it increases collagen synthesis by osteogenic cells. To test the effects of orthovanadate on bone nodule formation and mineralization, osteogenic cells were exposed to 5-50 microM orthovanadate or 10(-7) M insulin like growth factor-1 for 3, 7, and 21 days after plating. Exposure to orthovanadate produced differential effects on cellular proliferation and alkaline phosphatase activity following completion of the logarithmic growth phase, and on resultant bone nodule formation and mineralization by these populations. The effects of orthovanadate on osteogenic cultures were concentration dependent: 5 microM concentrations produced by a relatively large quantity of poorly mineralized matrix, while 30-50 microM concentrations produced a smaller quantity of heavily mineralized matrix. Thus, orthovanadate could possibly be used as a growth factor for bone, if administered at the critical dosage at the proper stage of the life cycle of the osteoblast. PMID- 9222446 TI - Lack of influence of cyclosporin A on levels of gingival procollagen types I and III mRNAs in rats of different ages. AB - Previous studies showed that gingival overgrowth following cyclosporin A (CsA) administration is not associated with an increase in interstitial collagen. It also was shown that CsA causes a significant decrease in collagen content within the gingival stroma. In order to determine whether this decrease is caused by down-regulation of collagen mRNA, the procollagen mRNA level in gingiva of young and old rats was measured correlated with the ratio of interstitial collagen to DNA in these regions. Hybridization of 32P-labelled cDNA probes for procollagen types I and III with total RNA extracted from the molar gingiva showed that administration of Csa did not change the steady-state levels of mRNAs for both procollagens in the gingiva of either young or old rats. The ratio of gingival interstitial collagen to DNA was significantly reduced in the CsA-treated animals (4.2 +/- 0.85) relative to the controls (7.8 +/- 1.6). It is concluded that the reduction in interstitial collagen following CsA treatment is not age-related, and is most probably caused by increased degradation rather by decreased biosynthesis. PMID- 9222448 TI - Low versus high pressure for in vitro determination of hydraulic conductance of human dentine. AB - The purpose of this study was to evaluate the effect of applied pressure and measurement time on the in vitro measurement of hydraulic conductance of human dentine. Dentine slices were prepared from 50 third molars. Water was forced through the slices under a constant hydrostatic pressure. Five pressures were tested: 1.3 kPa (n = 10), 13.3 kPa (n = 10), 26.6 kPa (n = 10), 40 kPa (n = 10) and 53.3 kPa (n = 10). The volume that went through the slices was recorded every 10 min for 3 h. The volume, the fluid flow rate and the hydraulic conductance under the five pressures were compared. The volume increased with time and pressure. The fluid flow and hydraulic conductance decreased with time under 13.3, 26.6 and 40 kPa, but remained constant under 1.3 and 53.3 kPa. Used of a low pressure (1.3 kPa) may permit water to pass through dentinal tubules without disturbing intratubular contents. Medium pressures (13.3, 26.6, 40 kPa) seemed to disturb tubule contents progressively, resulting in decreased fluid flow and therefore a decreased hydraulic conductance with time. Under these pressures, the calculated values of hydraulic conductance may be unreliable because they are time-dependent. High pressure (53.3 kPa) seemed to pack the tubule contents against intratubular resistances immediately, resulting in low fluid flows and low hydraulic conductances. PMID- 9222447 TI - Non-metric tooth crown traits of the Thai, Aka and Yao tribes of northern Thailand. AB - A survey was made of Thai tribe members, who cultivate rice paddies in the flatlands of northern Thailand, and of the Aka and Yao tribes, who farm with the slash-and-burn method in a mountainous region of northern Thailand. Plaster casts of the upper and lower jaws of tribe members were taken. Seventeen non-metric traits of their tooth crowns were classified and compared with other Mongoloid populations in various regions and periods. It was observed that the Thai tribe had the Sundadont characteristics, typical of South-East Asians, but the Aka and Yao tribe had more Sinodont than Sundadont characteristics, typical of North-East Asians. The regional and temporal variations of crown morphology in South-East Asia suggest earlier setting of the Thai tribe than the Aka and Yao tribes in this region. Moreover, comparison of the tooth morphological and linguistic classifications contradicts the traditional theory of the genealogy of the Thai language family. On the subject of the origin of modern South-East Asians, it is suggested that there has not been a gene flow of Sinodonty into Sundadonty of the principal ethnic groups in Neolithic South-East Asia. PMID- 9222449 TI - Chronic metabolic alkalosis, sucrose diet and dentine formation in young rats. AB - As acid-base status has an effect on bone formation and remodelling, chronic metabolic alkalosis was induced in 3-week-old rats for 6 and 7 weeks with 0.25 mol/1 of NaHCO3 in their drinking water to determine whether it has any effect on dentinogenesis in the molars. One group of rats was fed a high-sucrose diet and the other two a standard diet. The control groups had the same diets but drank distilled water. All the rats were injected with tetracycline to mark the onset of dentine apposition. The alkalotic effect of the NaHCO3 drinking water was confirmed by blood gas analysis at the end of the experiment. After death, tetracycline-marked dentine apposition was measured from sagittally sectioned mandibular molars. Chronic metabolic alkalosis did not affect dentine apposition in the groups with the high-sucrose diet, nor in the groups with the standard diet at 6 weeks, but reduced it significantly in first and second molars in 7 weeks at rats on the standard diet. A high-sucrose diet alone caused a greater reduction in the amount of dentine. The general growth of the rats was not affected in any of the groups. PMID- 9222450 TI - Immunocytochemical study of cathepsin L and rat salivary cystatin-3 in rat osteoclasts treated with E-64 in vivo. AB - The localization of cathepsin L and rat salivary cystatin-3 (RSC-3) in rat osteoclasts (rat femoral and alveolar bones) treated with or without E-64 (control) was examined immunocytochemically. In osteoclasts pretreated with E-64, immunoreactivity for cathepsin L was very weak extracellularly compared to that in the control osteoclasts. However, it was strong intracellularly. The localization of RSC-3 was unclear in the control osteoclasts, while in E-64 treated osteoclasts, both the clear zone and ruffled border areas showed a very strong immunoreaction. At the electron-microscopic level, in normal osteoclasts, numerous immunoreaction products for cathepsin L were found extracellularly in the bone matrix under the ruffled border, while few intracellular products were observed. In contrast, in the E-64-treated osteoclasts, only a few immunoreaction products were found extracellularly, while intracellularly cathepsin L was found in numerous endosome-lysosomal vacuoles. In the immunoreaction for RSC-3, the cytoplasm of the ruffled border was positive, and the tips of the RSC-3-positive ruffled border appeared to enter deeply into the bone matrix. Intracellularly, the granular reaction products of RSC-3 were found in the vacuoles (probably autophagolysosomes). Thus, in E-64-treated osteoclasts, inhibition of the extracellular release of cathepsin L was demonstrated. In addition, intralysosomal accumulation of RSC-3 and deep penetration of the RSC-3-positive ruffled border into the bone matrix were found. These findings suggest that RSC-3 is associated with the inhibition of cathepsin L in both the lysosomes (in the osteoclasts) and bone matrix. PMID- 9222451 TI - Influence of fluoride co-cystallized with sugar on caries development in desalivated rats. AB - Previous studies have shown that fluoride (F) administered concomitantly with sucrose in drinking water, in the diet, or alone is an effective cariostatic agent. The purpose of the present study was to determine the minimum concentration of F co-crystallized with sugar that may be used to prevent dental caries in rats subjected to a severe cariogenic challenge. Desalivated Sprague Dawley rats, infected with Streptococcus sobrinus, were placed in a Konig-Hofer programmed feeder. The rats received 17 meals daily for 21 days as follows: group (1) sucrose and sterile distilled water (s.d.w.); (2) sucrose and 1 part/10(6) F water; (3) 1 part/10(6) F-sucrose and s.d.w.; (4) 4 parts/10(6) F-sucrose and s.d.w.; (5) 8 parts/10(6) F-sucrose and s.d.w.; (6) sucrose and 10 parts/10(6) F water. Essential nutrition was administered by gavage. At death, blood was collected from each animal and one-half of the lower jaw was sonicated in 0.9% saline solution for microbial assessment and F analysis. Keyes smooth-surface and sulcal caries scores were significantly lower in the groups that received 10 parts/10(6) F-water and 8 parts/10(6) F-sucrose than in all other groups. The F concentration in the jaw suspension and plasma were significantly higher in the 10 parts/10(6) F-water and the 8 parts/10(6) F-sucrose groups than in all other groups. The total cultivable flora and Strep. sobrinus populations were lowest in F groups but this did not reach statistical significance. It is concluded that 8 parts/10(6) F co-crystallized with sucrose reduces the cariogenic potential of sugar as effectively as 10 parts/10(6) F in water, that is as little as 1 part/10(6) in sucrose has a significant effect, and that this cariostatic action is related to the amount of F in the oral environment. PMID- 9222452 TI - Effects of remote deep somatic noxious stimuli on a jaw reflex in man. AB - The effects on a inhibitory jaw reflex of activating deep somatic afferent nerves in a remote part of the body (the arm) were studied in 13 humans. Electromyographic recordings were made from the active masseter of the long latency (mean 42.0 +/- 1.1 ms) inhibitory reflex evoked by electrical stimulation of the upper lip. Immediately after a 1-min conditioning period during which the participants compressed a hand-held spring once a second while ischaemia was produced in the arm with an inflated pneumatic cuff, the magnitude of the inhibitory reflex decreased significantly (by 43%). The reflex recovered within 5 min to a magnitude that was not significantly different from its pre-conditioning value. The arm exercise or the ischaemia alone produced no significant changes in the reflex. Furthermore, neither of these last two conditions was reported to be painful, whereas the ischaemic exercise produced pain in all but one participant. It is concluded that activation of remote nociceptive, but not non-nociceptive, deep somatic nerves can modulate jaw reflexes in man. PMID- 9222453 TI - Automated image analysis applied to the odontoblast-predentine region in undemineralized sections of human permanent third molars. AB - A computerized histomorphometric analysis was made by Karnovsky-fixed, hydroxethylmethacrylate embedded and toluidine blue/pyronin-stained sections to determine: (1) the two-dimensional size of the coronal odontoblasts given by their cytoplasm:nucleus ratio; (2) the ratio between the number of coronal odontoblasts and dentinal tubules; and (3) the relation between odontoblast size and adjacent predentine. All conditions were measured in relation to three well defined sectioning profiles of the dentinal tubules. The sections were randomly taken from 10 unerupted and erupted third-molar crowns. Sixty-three photomicrographs (x100), equally distributed among the three sectioning profiles, were scanned in a high-resolution scanner to produce images for the analysis. After initial user interaction for the description of training classes on one image, an automatic segmentation of the images with respect to odontoblast cell nuclei, cytoplasm and background was computed by statistical discriminant analysis. In longitudinal profiles of the dentinal tubules the cytoplasm:nucleus ratio in erupted teeth was 3.1 +/- 0.54, and the mean of the odontoblast cell:dentinal tubule ration was 1.19 +/- 0.20. Analysis of cytoplasm:nucleus ratio and the adjacent predentine in relation to the chosen sectioning profiles disclosed that there was less variation in the predentine measurements in the longitudinal sections. Thus, in future two-dimensional studies of the odontoblast predentine region only longitudinal sectioning profiles should be analysed. The use of advanced image processing on undemineralized tooth sections provides a rational foundation for further work on the reactions of the odontoblasts to external injuries including dental caries. PMID- 9222454 TI - Effects of intrapartum hydrotherapy on labour related parameters. AB - The use of birthing pools during labour is increasing in the United Kingdom. This is without good scientific evidence of their efficacy or safety. To further investigate the value and safety of intrapartum hydrotherapy, an historical cohort study was performed in a District General Hospital in Liverpool. The study group consisted of 100 women of low obstetric risk who used the birthing pool at some stage during their labours and the control group consisted of 100 women who were matched in terms of age, parity and obstetric history but laboured and delivered in air. The main outcome measures were operative delivery rates, duration of labour, analgesic requirements, perineal trauma and Apgar scores at 1 and 5 minutes. The results showed that nulliparas who used the birthing pool had significantly reduced operative delivery rates, a shorter second stage of labour, reduced analgesic requirements and a lower incidence of perineal trauma. In multiparas there were significant reductions in analgesic requirements. PMID- 9222455 TI - Intermittent auscultation for the intrapartum assessment of fetal well-being in Western Australia. AB - In May 1995, in response to a decision of the Perinatal and Infant Mortality Committee of Western Australia, a survey of Western Australian hospitals was performed to ascertain what policies were in use for the monitoring of the fetal heart rate in labour and what proportion of these hospitals had access to electronic monitoring by cardiotocography. A response was received from 96% of the surveyed hospitals. More than half the births in this State (13,950 of 25,238) were monitored in labour using intermittent auscultation as the primary test; 7.5% of Western Australian births each year occurred in hospitals in which electronic monitoring was not available. Fewer than 50% of hospitals had written protocols describing the method of auscultation of the fetal heart during labour, the indications to contact a doctor or the management of fetal distress. The protocols which did exist displayed considerable variation in the recommended frequency of intermittent auscultation. The lack of standard practice in this field probably results from uncertainties in the literature. Intermittent auscultation has not been subjected to rigorous scientific evaluation as a screening tool and guidelines documenting ideal auscultatory practices need to balance the precision of electronic monitoring and freedom from intervention. Based on this compromise and existing evidence, a protocol for intermittent auscultation in normal labour is proposed. PMID- 9222456 TI - Antenatal patterns of uterine activity in low-risk women: a longitudinal study. AB - There is a surprising lack of information on antenatal patterns of uterine activity in the normal obstetric population, with the majority of research having focussed on women at high-risk for preterm birth. We conducted a prospective longitudinal study to investigate patterns of uterine activity in women with singleton gestations at low-risk for preterm birth. Twenty pregnant women were recruited and their uterine activity was recorded using ambulatory tocodynamometry twice weekly throughout the latter half of pregnancy. The collected data were transmitted to a central receiving station for analysis. As gestation advanced there was a progressive increase in the median number of contractions detected per hour, peaking and stablizing at 37-40 weeks (median of 0 contractions/hour at 20-24 weeks rising to 5.4 contractions/hour at 37-40 weeks). In those women with uterine activity, contraction duration and amplitude of deflection significantly increased as gestation advanced. There was a progressive increase in the number of higher amplitude contractions throughout the third trimester. Increasing parity was not associated with increasing antenatal uterine contraction frequency. No association between normal daily physical activity and uterine contraction frequency was evident throughout gestation. In normal human pregnancy there is a steady, progressive increase in the frequency, duration and amplitude of antenatal uterine activity throughout the latter half of gestation. The uterine contractile profile alters from one of a low amplitude, low frequency pattern in the second trimester to a higher amplitude, higher frequency pattern at term. PMID- 9222457 TI - The relationship between gestational age and the incidence of classical caesarean section. AB - Improved neonatal survival has led to a rise in the number of Caesarean sections being performed in the presence of extreme prematurity. Many of these operations require an incision in the upper uterine segment with consequent ramifications for the management of any subsequent pregnancy. In this analysis of obstetric patients in a tertiary referral institution over a 9-year period, there was an overall Caesarean section rate of 18%. A classical incision was performed in 1% of all caesarean sections, but at 24 weeks' gestation, 20% of Caesarean sections were 'classical'. This frequency decreased to less than 5% at 30 weeks and less than 1% from 34 weeks' gestation. Most women having a classical Caesarean section at term had either a transverse lie or a major degree of placenta praevia. PMID- 9222458 TI - Peripartum cardiomyopathy. AB - Peripartum cardiomyopathy is an uncommon condition of unknown aetiology. Diagnosis requires exclusion of other causes of congestive cardiac failure and the demonstration of global ventricular dysfunction on echocardiography. Treatment consists od diuretics, vasodilators, digoxin and anticoagulants. Prognosis is related to recovery of ventricular function. The availability of cardiac transplantation has improved the outlook for those with persistent dysfunction. PMID- 9222460 TI - How well does epidemiological evidence hold for the relationship between smoking and adverse obstetric outcomes in New South Wales? AB - This study examines whether the established epidemiological relationship between cigarette-smoking exposure in pregnancy and adverse obstetric outcomes are confirmed in Australian data from the NSW Midwives Data Collection (MDC). These data were analysed to compare the obstetric complications and pregnancy outcomes between smoking and nonsmoking women confined in 1994. Results showed that smoking mothers had higher rates of antepartum haemorrhage due to placental abruption and placenta praevia. They were also at higher risk of giving birth to low birth-weight babies and preterm delivery. Infants born to smoking mothers were 17% more likely to be admitted to hospital special care nurseries or neonatal intensive care units. Moreover, the risk of reported perinatal death among babies of smoking mothers was 20% higher than babies of nonsmoking mothers. However, smoking during pregnancy was found to confer a protective effect against the development of pregnancy-induced hypertension. These results were compared with existing evidence from the literature. Published research reports on the corresponding smoking effects were identified to assess the consistency of evidence and typical risk ratios. Findings from the literature search showed a near-perfect concordance with the associations in the NSW MDC data. The paper documents the likely complications which might be prevented if smoking in pregnancy were eliminated. There remains a real need for effective programmes to reduce smoking prevalence in pregnancy. PMID- 9222461 TI - Balloon catheter with extra-amniotic saline instillation: a method of induction in pregnancies at 41 or more gestational weeks. AB - One hundred five women with a documented pregnancy of 41 weeks' gestation or more, admitted for induction of labour by balloon catheter with extra-amniotic saline instillation, were retrospectively compared to 196 women admitted in spontaneous labour at the same gestational age, with regard to mode of delivery. The success rate of the induction group was 97.1%. The mode of delivery did not differ significantly between the 2 groups. The Caesarean section rates were 11.4% in the induction group versus 9.7% in the spontaneous group. The mode of delivery after induction of labour by balloon catheter with extra-amniotic saline instillation and simultaneously commenced intravenous oxytocin infusion, in pregnancies of 41 weeks or more, is similar to that of spontaneous deliveries at the same gestational age. PMID- 9222459 TI - Survival and neonatal and neurodevelopmental outcome of 24-29 week gestation infants according to primary cause of preterm delivery. AB - A total of 189 infants of 24-29 weeks' gestation were born in a regional perinatal centre during a 2-year period. They were divided into groups according to the primary cause of preterm delivery: antepartum haemorrhage (n = 37, 20%), preeclampsia (n = 27), 14%), preterm premature rupture of membranes (n = 64, 34%), preterm labour (n = 27, 14%), chorioamnionitis (n = 16, 8%), other complications (n = 18, 10%). The perinatal mortality rate (PMR) was 286/1,000 of whom 44% were stillbirths. The 'other complication' group had the highest PMR due to a large number of intrauterine deaths, with no differences in neonatal mortality between the groups. Preeclampsia was associated with an increased risk of necrotizing enterocolitis and chorioamnionitis was associated with an increased risk of periventricular haemorrhage. Follow-up to at least 2 years was performed in 122 (97%) of survivors. Cerebral palsy occurred in 7%, while 18% had neurodevelopmental disability. No relationship was found between primary cause of preterm delivery and outcome. This information should be of value in counselling parents when preterm delivery is imminent. PMID- 9222462 TI - The metabolic profile of glucose tolerant women who have had large for gestational age babies. AB - The possibility has been raised that women who have had large for gestational age infants, while glucose tolerant during pregnancy by conventional testing, may still have a subtle abnormality of carbohydrate metabolism. We have examined, some time after the completion of a pregnancy, the fasting levels of glucose, insulin and lipids in a group of women, glucose tolerant during pregnancy, who had a large for gestational age infant compared to a very carefully matched control group of women who had an appropriate for gestational age infant. No significant differences were found. These findings suggest that women who have a large for gestational age infant do so for a variety of reasons not related to maternal carbohydrate metabolism. PMID- 9222463 TI - Antenatal screening for HPA-1a by flow cytometry. AB - Pregnant women who attended antenatal clinics at King George V Hospital, the Birth Centre or were referred by obstetricians from February 19 July, 1996 were screened for the platelet antigen HPA-1a by flow cytometry. Forty out of 2,300 (1.7%) were found to be negative for this antigen. Of the 28 women followed throughout their pregnancy, none developed antibody to HPA-1a. Platelet counts performed on samples from 17 babies born to 17 of these mothers were all normal. This study proves the simplicity and rapidity of flow cytometry for platelet antigen screening. The results were comparable with the Solid Phase Red Cell Adherence (SPRCA) method and with PCR. The lack of a plentiful supply of specific antibody and the rarity of fetomaternal alloimmune thrombocytopenia (FMAIT) argue against the introduction of routine screening for maternal HPA-1a status at the present time. PMID- 9222464 TI - Gestational diabetes mellitus; resource utilization and costs of diagnosis and treatment. AB - The recommendation to test every woman for gestational diabetes mellitus (GDM) has a defined cost. The management of women diagnosed with GDM will use additional health resources. This examines the cost and resource utilization of a consecutive group of women diagnosed over a 1-year period. The cost of testing a woman for GDM is around $10.00 with slight variations depending on the testing procedure. The annual cost of testing in NSW would be less than 1 million dollars. Women diagnosed with GDM used the resources of a diabetes education centre for an average of 2.8 hours and attended for 3.4 (2.3) medical visits. Insulin was required by 18.7% of the women for 9.7 (4.7) weeks using 47.7 (21.2) units each day. Testing women for GDM is a low-cost item. Managing a woman diagnosed with GDM may cost several hundred dollars. Cost reductions could be made by reducing the amount of insulin used and by avoiding hospitalization. PMID- 9222465 TI - Twin-reversed arterial perfusion (TRAP) sequence: case reports and review of literature. AB - The twin reversed-arterial-perfusion (TRAP) sequence found in monozygotic twins is a consequence of primary or secondary cardiac development disruption and direct arterioarterial and venovenous placental anastomoses. Associated findings include the presence of a single umbilical artery (66%) and chromosomal abnormalities in the acardiac twin (33%). Morphological abnormalities in the acardiac twin are consistent with perfusion of tissues supplied by the common iliac and lower branches of the aorta with deoxygenated blood. The pump or donor twin may develop cardiac failure because of the anomalous perfusion circuit. Polyhydramnios is significantly associated with presence of renal tissue in the acardiac twin. An acardiac pump twin weight ratio (> 50%) is associated with the development of polyhydramnios and preterm labour. Identified high-risk factors for poor obstetrical outcome include: acardiac anceps, polyhydramnios, acardiac twin with ears, and pump twin cardiac failure. Management options include elective termination, observation (serial cardiotocography (CTG), ultrasonography and echocardiography) and selective nonsurgical interventions (indomethacin, digitalis, tocolysis). Additionally, surgical interventions (hysterotomy with selective delivery of the acardiac twin or ligation of the acardiac twin's umbilical cord), and ultrasound-guided embolization of the acardiac twin's umbilical artery with absolute alcohol, platinum coils, or thrombogenic coils have been reported. The most appropriate interventions for the various clinical presentations of this disorder are as yet undetermined, and conservative nonintervention is often appropriate. Long-term follow-up data on surviving pump twins are lacking. It is anticipated that centres with active study protocols for these conditions will best serve patient care and clinical research needs. PMID- 9222466 TI - Medicolegal matters involving a major obstetric and gynaecological teaching hospital. AB - Review of medicolegal files held by the Royal Women's Hospital. Melbourne confirms that during the last 25 years there has been a marked increase in the number of claims for compensation brought by patients who believe that the care they received was inadequate; thus 4 claims for compensation were received during the first 5 years of the study period and 29 claims in the last 5 years. Complaints about service provision resulted in a claim for compensation in 29.7% of cases in which the dissatisfied client was represented by a legal firm and in 6.25% of cases where the initial approach was made through the Health Services Commission. One half of all claims for compensation were received in response to perceived complications of birth, surgery or treatment of a premature baby. PMID- 9222467 TI - The levonorgestrel-releasing intrauterine device: a wider role than contraception. AB - The Levonorgestrel-releasing intrauterine device (LNG IUD) provides excellent contraception; it may reduce the rate of pelvic inflammatory disease (PID) and ectopic pregnancy compared to other 'modern' copper releasing IUDs; it can safely be used in the puerperium for breast-feeding mothers, and it significantly reduces menstrual blood loss and pain. While it was developed primarily as a contraceptive, its potential role in managing heavy and painful menstruation and the symptoms of the climacteric may eventually be just as important. Amongst developed countries New Zealand and Australia have some of the highest hysterectomy rates. By the age of 50 years 1 in 4 women in New Zealand and 1 in 5 women in Australia will have had a hysterectomy (A,B). In New Zealand 90% of these are performed for heavy menstrual bleeding and fibroids (A). The LNG IUD has been shown to be effective treatment for both these conditions and its introduction to New Zealand and Australia would offer women an additional choice beyond surgery. PMID- 9222468 TI - Outpatient hysteroscopy--problems. AB - Outpatient hysteroscopy is now a routine procedure, but difficulties may be encountered. It was decided to assess the problems found in one practice over a period of time. Between August 1, 1989 and July 31, 1996 there were 1,080 examinations performed by the author; pain was sometimes significant, but already has been well covered in the literature. The other important problems were vasovagal reactions and failure to complete an adequate examination. This report emphasizes these problems. Being aware of potential problems allows one to anticipate them and often allows the procedure to be carried out successfully. At the very least, this awareness should save the patient major medical problems or discomfort. PMID- 9222469 TI - Saline infusion sonohysterosalpingography, an underutilized technique. AB - Saline infusion sonohysterography (SIS) is an important gynaecological diagnostic tool which is little used in Australia. We herein report the findings in 60 women referred for SIS, the procedure being uneventfully performed in 55. Forty-nine of the 60 referrals were for investigation of abnormal uterine bleeding. The technique described allows examination of the uterine cavity and the Fallopian tubes. In 26 of the patients information was obtained which improved or altered the diagnosis made on B mode and colour Doppler ultrasound. Unlike hysteroscopy, SIS is always performed as an outpatient procedure, appropriate disinfection procedures are relatively simple to implement (1), and vasovagal reactions are rare. Hysteroscopy was avoided in 11 patients, there were other benefits in 4 patients, and in only 1 of 16 patients did the hysteroscopy findings differ with SIS. If diagnostic pitfalls are avoided by careful attention to detail, SIS offers a powerful new gynaecological investigative tool in the investigation of bleeding disorders (including menorrhagia, intermenstrual and postmenopausal bleeding), uncertain endometrial findings on vaginal ultrasound, infertility, and in the investigation of congenital and acquired uterine abnormalities. PMID- 9222470 TI - Hysterectomy and endometrial ablation in New South Wales, 1981 to 1994-1995. AB - This study analyses the New South Wales hospital data on hysterectomies from 1981 to 1994-1995, and on endometrial ablations since 1991. The hysterectomy rate declined by about 16% during 1981-1991 and has risen since; the endometrial ablation rate has increased by 28% between 1991 and 1994-1995. Other findings indicate a trend towards older mean age at operation, a swing to vaginal hysterectomy with or without laparoscopy, a shift to private hospitals, and a dramatic decline in length of hospital stay. The majority of endometrial ablations were performed on a day-only basis. Immigrant and Aboriginal women experienced lower hysterectomy rates. Endometrial ablation techniques introduced in the late 1980s, as an alternative to hysterectomy for dysfunctional uterine bleeding, have had a major impact on hysterectomy rates; without these techniques the rates would be much higher. PMID- 9222471 TI - The safety of laparoscopy performed by direct trocar insertion and carbon dioxide insufflation under vision. AB - The records of 6,173 laparoscopies performed by specialist gynaecologists in the course of routine gynaecological care using the technique of direct insertion of the umbilical trocar and insufflation of carbon dioxide under vision were reviewed to ascertain the incidence of serious complications. A review of the published literature on laparoscopy methodology was also undertaken to complement the data obtained from this study. The nature of the records precluded accurate assessment of both indications and minor complications. There were 4 perforating bowel injuries (0.06%) requiring laparotomy (s small intestine, 2 large intestine). There were no cases of major vascular injury or gas embolus necessitating surgical or resuscitative measures. On 3 of the 4 occasions where bowel injury occurred the patients had undergone prior abdominal surgery and had midline vertical subumbilical incisions. Review of the published literature demonstrated bowel or vessel perforation rates (requiring laparotomy or resuscitation) of 1 in 1,000 regardless of whether the method of gaining peritoneal access was open (Hasson) technique, Verres needle insufflation, or direct trocar. Direct trocar insertion may reduce the risk of gas embolism by insufflating only after intraperitoneal replacement has been confirmed, moreover it allows immediate recognition and rapid treatment of major blood vessel laceration, both of which have been identified as being crucial in reducing laparoscopy associated mortality. When compared to other available methods of gaining peritoneal access for laparoscopy, direct trocar insertion followed by insufflation of carbon dioxide under vision can be performed with the same degree of safety for the patient. It is simply wrong to deduce from the available data that one particular technique of gaining peritoneal access is superior to another. Each have their individual advantages and disadvantages and similar morbidity when performed by experienced operators with appropriate indications. In light of this observation, each alternative should be considered by the individual surgeon to assess which would best suit his or her operating technique and the particular circumstance of each patient. Indeed preference should be given to the method with which the surgeon is most comfortable or with which he or she has the most experience. PMID- 9222472 TI - Vault haematoma following laparoscopic hysterectomy. AB - Thirty consecutive patients underwent transabdominal ultrasound scanning on day 2 postoperatively in order to provide data on the incidence of vaginal vault haematoma following laparoscopic hysterectomy. Details of postoperative morbidity, both inpatient and after discharge, were recorded. Results support the view that there is no significant association between the presence of vaginal vault haematoma (73%) and the incidence of posthysterectomy febrile morbidity (16.7%). Furthermore the incidence of vault haematoma after laparoscopic hysterectomy is comparable to literature figures for both abdominal and vaginal hysterectomy, whilst that of febrile morbidity is at least equivalent if not reduced for the laparoscopic approach. We believe this provides further evidence confirming the safety of the laparoscopic approach to hysterectomy. PMID- 9222473 TI - Pre and intraoperative diagnosis of ovarian tumours: how accurate are we? AB - In the assessment of malignant potential of ovarian tumours, frozen section has been found to be accurate in 97.1% (168 of 173) of cases. The positive predictive value of frozen section in the diagnosis of a malignant lesion was 100% (34 of 34). Errors were mainly made in the diagnosis of borderline tumours with a predictive value of 87.5% (7 of 8). The negative predictive value was 98.4% (127 of 129). Frozen section however, was less accurate in the diagnosis of specific histological type with an accuracy rate of 91.9% (159 of 173). Macroscopic features were found to be useful in the intraoperative prediction of malignant potential. Completely cystic tumours were benign in 96.4% (108 of 167) of cases. Solid/cystic tumors were malignant in 69% (27 of 38) of cases. Completely solid tumours were malignant in 56% (9 of 16) of cases. Frozen section in completely cystic tumours only marginally improved the clinical macroscopic diagnosis of malignancy. The sensitivity and specificity of ultrasound scan in the diagnosis of malignant/borderline tumours were 82% and 86% respectively. The false negative rate of 7% makes laparoscopic excision of unsuspected malignant ovarian cyst a significant possibility. The predictive value of ultrasound scan in the diagnosis of malignant ovarian tumour was 62% (26 of 42). In the preoperative assessment of malignant potential of ovarian tumours, this study shows that ultrasound scan has a high false positive and a significant false negative rate. Careful intraoperative assessment of gross features and the use of frozen section especially in those with solid/cystic and solid tumours will help achieve a high accuracy rate in the assessment of ovarian tumours. PMID- 9222474 TI - High-dose medroxyprogesterone acetate for the treatment of dysfunctional uterine bleeding in 24 adolescents. AB - The objective of the study was to evaluate the effectiveness of high-dose oral medroxyprogesterone acetate therapy in the management of excessive dysfunctional uterine bleeding in adolescents. The study group consisted of 24 adolescents who were hospitalized with the diagnosis of excessive uterine bleeding and anaemia. Oral medroxyprogesterone acetate tablets were administered at a total dose of 60 120 mg during the first day of admission and 20 mg per day for the following 10 days. The blood loss was reduced to acceptable levels in all patients, and actually stopped in 6 (25%) within the first 24 hours of the treatment; bleeding ceased in 29.2%, 20.8% and 25% on the second, third and fourth days respectively. Significant correlation was identified between the initial haemoglobin concentration and the time required for cessation of bleeding (r2 = 0.5, p = 0.001). Rapid saturation of the endometrium with progestogens seems to be an highly effective mode of treatment for excessive dysfunctional uterine bleeding in adolescents. PMID- 9222475 TI - The predictive value of the sperm-cervical mucus interaction test on the outcome of in vitro fertilization and ovulation induction combined with intrauterine insemination. AB - We investigated the effect of the sperm-cervical mucus penetration tests (SPT) on the fertilization rate (FR) and pregnancy rate (PR) in patients treated with either in vitro fertilization (IVF) or ovulation induction combined with intrauterine insemination (OI + IUI). Infertile couples where the women had normal ovarian function and a normal pelvis at laparoscopy and her partner had normal seminology who had failed at least 2 SPTs were treated with either IVF or OI + IUI. These patients were compared with similar couples in whom SPTs were satisfactory (SPT/ve). Group A (SPT+ve) consisted of 46 patients who underwent 78 treatment cycles of IVF and Group B (SPT-ve) comprised 31 patients who underwent 35 IVF cycles. Group C (SPT/ve) consisted of 39 patients who underwent 84 treatment cycles with OI + IUI, and Group D (SPT-ve) consisted of 15 patients who underwent 37 cycles with the same treatment. In patients treated with IVF, the FR and PR per embryo transfer were 77.0% and 20.0% respectively in Group A, and 64.0% and 22.6% respectively in Group B. The difference in FRs was statistically significant (p > 0.001) but there was no difference in the PRs. In patients treated with OI + IUI, the PR per cycle were 22.0% in Group C and 16.2% in Group D. These results indicate that SPT failure was associated with a lower FR in IVF but this did not affect the PRs. Similarly there was no difference in PRs following OI + IUI. PMID- 9222476 TI - A case of complete iatrogenic amnioreduction at 20 weeks' gestation. PMID- 9222477 TI - Aortic compression in massive postpartum haemorrhage--an old but lifesaving technique. PMID- 9222478 TI - An unusual case of acute postpartum broad ligament haematoma. PMID- 9222479 TI - Intractable pelvic pain following Filshie clip application. PMID- 9222480 TI - Coital tear: a rare cause of secondary peritonitis. AB - A rare cause of secondary peritonitis due to coital tear is presented. The correct diagnosis can be made by a detailed history and gynaecological examination. Prompt surgical management is mandatory to prevent grave prognosis. PMID- 9222481 TI - Re: Fearing the worst--why do pregnant women feel 'at risk'? PMID- 9222483 TI - The incidence of perinatal mortality associated with hyperglycaemia in pregnancy. PMID- 9222482 TI - The effect of epidural bupivacaine on the fetal electrocardiogram. PMID- 9222484 TI - Bone biology. AB - Bone is a metabolically active and highly organized tissue consisting of a mineral phase of hydroxyapatite and amorphous calcium phosphate crystals deposited in an organic matrix. Bone has two main functions. It forms a rigid skeleton and has a central role in calcium and phosphate homeostasis. Bone modelling is the process associated with growth and re-shaping of bones in childhood and adolescence. This is distinguished from bone remodelling, which describes the lifelong process whereby skeletal tissue is continually being resorbed and replaced in order to maintain skeletal integrity, shape and mass. Bone remodelling is controlled by systemic hormones and cytokines and is an integral part of the calcium homeostatic system. The maintenance of a normal, healthy skeletal mass depends on interactions between osteoblasts, osteoclasts and constituents of the bone matrix to keep the process of bone resorption and formation in balance. The factors, local and systemic, which regulate these processes are discussed. PMID- 9222485 TI - The epidemiology and pathogenesis of osteoporosis. AB - Osteoporosis is an increasing health care concern as populations age throughout the developed and developing world. The social and economic costs of osteoporosis are due to its clinical outcome of fracture which increases exponentially with age. This review will highlight some of the key epidemiological aspects of osteoporosis incorporating areas of more recent interest. These include the definition; the magnitude of the problem encompassing differing incidence and prevalence patterns of both low bone mass and fracture in different cultural groups; the social consequences of fracture, including economic costs, morbidity and mortality; the evaluation of fracture risk, including the role of bone density, bone quality and the risk of falling; as well as an overview of some of the factors involved in determining low bone mass. Bone mineral density (BMD) is the most easily measured and accurate predictor of fracture risk. For any individual, BMD is the combination of their peak bone density and subsequent bone loss, both of which are influenced by genetic, hormonal and environmental factors. An understanding of key issues relating to this important disease may lead to earlier detection of the individual at high risk for fracture and rational approach to prevention and management. PMID- 9222486 TI - The management of osteoporosis. AB - The management of osteoporosis used to centre upon investigation and treatment of patients with fracture. The spectrum has now widened to include the detection of patients at high risk of fracture before a fracture occurs. This is best achieved by consideration of clinical risk factors and the selective use of bone densitometry. The frequency of osteoporotic fractures in elderly women is such that detailed investigation of such patients is often not necessary unless the patient's bone density is outside the normal range for age. When bone density is inexplicably low, secondary causes of osteoporosis should be sought by appropriate investigations. Fracture prevention involves a correction of lifestyle factors (stopping smoking, moderating alcohol intake etc.) and achieving a total calcium intake of 1-1.5 g/day. The first line for pharmacological intervention is hormone replacement therapy because of its proven efficacy and the extensive data available documenting its safety. The bisphosphonates have comparable effects on bone density and fractures in studies extending for up to 3 years, and are increasingly used, particularly in older patients and those reluctant to take hormone replacement therapy. Other available agents have significant drawbacks, either with respect to side-effects or inconsistent documentation of efficacy and should be used only in special circumstances. PMID- 9222488 TI - The skeletal effects of primary hyperparathyroidism. AB - Primary hyperparathyroidism (PHPT) is a common endocrine disorder which occurs most frequently in post-menopausal women and is characterized by mild, stable, and often asymptomatic hypercalcaemia. Chronic parathyroid hormone excess stimulates bone remodelling by inducing production by osteoblasts of soluble factors which stimulate both bone formation and osteoclastic bone resorption. Studies of bone mineral density (BMD) in PHPT suggest that bone loss is accelerated, leading to osteopenia, particularly at sites of cortical bone. Studies of fracture incidence in PHPT have produced conflicting results. Interventional studies have demonstrated that both parathyroid adenomectomy and estrogen replacement therapy increase BMD in patients with PHPT. Patients with PHPT should undergo BMD measurement, and receive treatment designed to stabilize bone mass if there is evidence of either osteopenia or accelerated bone loss. PMID- 9222487 TI - Secondary osteoporosis. AB - Secondary osteoporosis is diagnosed when there is a well-established disease related risk factor for fracture or low bone mass. Secondary osteoporosis is associated with a substantial minority of osteoporotic fractures in women perhaps with a majority of osteoporotic related fractures in men. This chapter does not review all the possible causes of low bone mass and fractures but picks out some of the more important causes of, with an emphasis on the main iatrogenic cause, that is corticosteroid induced osteoporosis. It also highlights some of the possible causes which could be avoidable. Where appropriate the methods of prevention and treatment of secondary osteoporosis are reviewed. PMID- 9222489 TI - Paget's disease of bone: clinical, pathogenetic and therapeutic aspects. AB - Paget's disease of bone is a focal disorder of bone remodelling due to abnormally increased osteoclast-mediated bone resorption. It rarely presents before the age of 35 years and its prevalence increases with age affecting 2-5% of the population above 50 years, making Paget's disease the most common skeletal disorder after osteoporosis. Its aetiology is not known but available evidence favours an infection of genetically predisposed individuals with a paramyxovirus. Affected bones change in shape, size and direction causing considerable morbidity but the majority of patients are asymptomatic. In Paget's disease increased bone resorption is tightly linked to increased bone formation which are reflected in the proportional increases in biochemical indices of bone turnover. Because the primary abnormality lies in the osteoclasts, inhibitors of bone resorption are used for its therapy and bisphosphonates are currently the treatment of choice. These decrease bone turnover effectively and their effect lasts after stopping treatment. With available bisphosphonates clinical and biochemical, long-lasting, remissions can be obtained in the majority of patients. Symptomatic disease and preparation for orthopaedic surgery are no longer the only indication for treatment, but asymptomatic patients with localizations at sites likely to induce complications should be considered candidates for bisphosphonate therapy. PMID- 9222490 TI - Osteomalacia. AB - Osteomalacia is a generalized bone disorder characterized by impairment of mineralization, leading to accumulation of unmineralized matrix or osteoid in the skeleton. The classical clinical features of osteomalacia include musculoskeletal pain, skeletal deformity, muscle weakness and symptomatic hypocalcaemia. In childhood the features of osteomalacia are accompanied by rickets, with widening of the epiphyses and impaired skeletal growth. The major cause of osteomalacia is vitamin D deficiency, which is most often due to reduced cutaneous production of vitamin D in housebound elderly people, immigrants to Northern countries and women who adopt strict dress codes which prohibit exposure of uncovered skin. Vitamin D deficiency osteomalacia may also occur with malabsorption, liver disease and anticonvulsant therapy. Less commonly, osteomalacia may result from abnormal vitamin D metabolism, resistance to the action of vitamin D, hypophosphataemia or toxic effects on osteoblast function. PMID- 9222491 TI - Renal osteodystrophy. AB - Renal osteodystrophy is a general complication of chronic renal failure and end stage renal disease. The nature of renal osteodystrophy has changed since osteomalacia due to aluminum intoxication has become less prevalent. Osteomalacia has been replaced by the adynamic bone disorder. Suppression of osteitis fibrosa, calcitrol and control of secondary hyperparathyroidism has been shown to produce the adynamic bone disorder. Thus, many other factors besides secondary hyperparathyroidism and calcitrol deficiency contribute to the pathogenesis of renal osteodystrophy. Some of these factors, according to our current state of knowledge, are discussed in this chapter along with the presentation and treatment of renal osteodystrophy. PMID- 9222492 TI - Osteogenesis imperfecta and other heritable disorders of bone. AB - This chapter summarizes the many recent advances in our understanding of the principal heritable disorders of bone. In the course of little more than a decade many diseases that were recognizable only by their clinical and radiological features have become explicable in molecular terms. Large numbers of mutations of the genes coding for collagen, for alkaline phosphatase, for the cell surface receptors for parathyroid hormone and for calcium, and for a number of other proteins, are recognized. The chapter covers the many variants of osteogenesis imperfecta, the most common heritable cause of fractures. It also covers osteopetrosis, hypophosphatasia, pseudohypoparathyroidism (with Albright's hereditary osteodystrophy), familial benign hypercalcaemia, autosomal dominant hypocalcaemia and the molecular causes of some chondrodysplasias. PMID- 9222494 TI - A strategy of tRNA recognition that includes determinants of RNA structure. AB - Recognition of tRNAs by aminoacyl tRNA synthetases establishes the connection between amino acids and anticodon triplets of the genetic code. Although anticodons and nucleotides adjacent to the amino acid attachment site are generally important, the tertiary structural framework of tRNAs has recently been implicated to have a role in tRNA recognition. A G15:G48 tertiary hydrogen base pair of E. coli tRNA(Cys) is important for recognition of the tRNA by cysteine tRNA synthetase. This base pair is proposed to consist of N2:N3, rather than N1:O6, hydrogen bonds. The reproduction of the hydrogen pairing scheme of tRNA(Gly). This reproduction required an A13:A22 mismatch in the dihyrouridine stem. To determine if A13:A22 is a determinant of the structural features of G15:G48, we investigated the A15:U48 and A15:A48 variants of tRNA(Gly) which harbored specific substitutions of A13:A22. We show here that introduction of A13:A22 to both tRNA frameworks confers structural features similar to those of G15:G48 in E. coli tRNA(Cys). These structural features are accompanied by efficient recognition of both tRNAs by cysteine tRNA synthetase. Substitution of A13:A22 with U13:A22 alters the structural features at 15:48 and impairs tRNA recognition. The dependence on A13:22 for tRNA recognition has a distinct similarity to that of E. coli tRNA(Cys) and to that of the G15:G48 variant of tRNA(Gly). The results have implications for the design and manipulation of RNA structural elements as the basis for tRNA recognition. PMID- 9222493 TI - Strategy for RNA recognition by yeast histidyl-tRNA synthetase. AB - Histidine aminoacylation systems are of interest because of the structural diversity of the RNA substrates recognized by histidyl-tRNA synthetases. Among tRNAs participating in protein synthesis, those specific for histidine all share an additional residue at their 5'-extremities. On the other hand, tRNA-like domains at the 3'--termini of some plant viruses can also be charged by histidyl tRNA synthetases, although they are not actors in protein synthesis. This is the case for the RNAs from tobacco mosaic virus and its satellite virus but also those of turnip yellow and brome mosaic viruses. All these RNAs have intricate foldings at their 3'-termini differing from that of canonical tRNAs and share a pseudoknotted domain which is the prerequisite for their folding into structures mimicking the overall L-shape of tRNAs. This paper gives an overview on tRNA identity and rationalizes the apparently contradictory structural and aminoacylation features of histidine-specific tRNAs and tRNA-like structures. The discussion mainly relies on histidylation data obtained with the yeast synthetase, but the conclusions are of a more universal nature. In canonical tRNA(His), the major histidine identity element is the 'minus' 1 residue, since its removal impairs histidylation and conversely its addition to a non-cognate tRNA(Asp) confers histidine identity to the transplanted molecule. Optimal expression of histidine identity depends on the chemical nature of the -1 residue and is further increased and/or modulated by the discriminator base N73 and by residues in the anticodon. In the viral tRNA-like domains, the major identity determinant -1 is mimicked by a residue from the single-stranded L1 regions of the different pseudoknots. The consequences of this mimicry for the function of minimalist RNAs derived from tRNA-like domains are discussed. The characteristics of the histidine systems illustrate well the view that the core of the amino acid accepting RNAs is a scaffold that allows proper presentation of identity nucleotides to their amino acid identity counterparts in the synthetase and that different types of scaffoldings are possible. PMID- 9222495 TI - Context dependent RNA-RNA recognition in a three-dimensional model of the 16S rRNA core. AB - A 3-D model of the core of the 16S rRNA of Escherichia coli containing 328 residues has been built in the protein map derived from neutron scattering data with the help of all the available phylogenetic, biochemical, and cross-linking data. The three pseudoknots of the 16S-core cluster, through the arrangement of complex three-, four- and five-way junctions, around the neck and at the subunit interface. The roles in assembly, initiation or elongation of the three pseudoknots in ribosomal dynamics are emphasized. The 530-loop, localized on the periphery of the 30S particle, could be built with and without a pseudoknot independently of the state of the particle. The pseudoknot of the central domain controls the dynamics of an helix connected to the subunit interface which could trigger some mechanism during translation. The process of the model construction is compatible with a folding scenario in which the 5'-terminal pseudoknot controls the assembly of the central junction and the subsequent folding of the 3'-major domain. The modelling, together with the phylogenetic analysis and the experimental data, point to several potential RNA-RNA contacts which depend on the structural and sequence context in which they occur. PMID- 9222496 TI - Modulation of RNase H activity by modified DNA probes: major groove vs minor groove effects. AB - We have previously prepared ribozyme mimics and chemical nucleases from modified DNA containing pendant bipyridine and terpyridine groups. The ability of these modified DNA probes to support RNase H cleavage of complementary RNA is described. DNA/RNA duplexes were formed using DNA probes designed to deliver metal complexes via either the major groove or the minor groove of the duplex. The duplexes were treated with Escherichia coli RNase H. Modifications in the major groove produced the same RNA cleavage pattern as unmodified DNA probes. However, minor groove substituents inhibited RNA cleavage over a four-base region. Comparison was made with a DNA probe containing a 2'-OMe modification. Our results support enzyme binding in the minor groove of a DNA/RNA duplex. We do not observe cleavage directly across from the modified nucleoside. The RNA cleavage efficiency effected by RNase H and a DNA probe decreases as follows: unmodified DNA > or = C-5 modified DNA >> c2'-modified DNA > C1'-modified DNA. Results with 28-mer RNA substrates roughly parallel those obtained with a 159-mer RNA target. The differences observed between low and high MW RNA substrates can be explained by a much higher enzyme-substrate binding constant for the high MW target. PMID- 9222497 TI - Recognition of RNA by triplex formation: divergent effects of pyrimidine C-5 methylation. AB - In DNA triple helices, methylation at C-5 of thymine or cytosine is reported to have similar stabilizing effects for both bases. Here we show, however, that methylation of the same positions in RNA triplexes has distinctly different effects than in DNA. We have previously described the use of circular triplex forming RNA oligonucleotides to recognize RNA sequences. Here it is shown that addition of C-5 methyl groups to uracils in these compounds very significantly increases not only affinity but also sequence selectivity in binding a purine rich RNA target, as measured by thermal denaturation with various target RNAs. Surprisingly, however, addition of C-5 methyl groups to cytosines actually decreases affinity in binding RNA, while the same substitution in DNA is thermally stabilizing. Possible sources of this divergent behavior are discussed. A synthesis of 5-methylcytidine ribonucleoside 2'-O-silyl-3'-O-phosphoramidite is also described. PMID- 9222498 TI - Application of a 5'-bridging phosphorothioate to probe divalent metal and hammerhead ribozyme mediated RNA cleavage. AB - This paper describes the preparation and application of a chimeric DNA/RNA oligonucleotide that contains a single 5'-bridging phosphorothioate linkage adjacent to a ribonucleotide and embedded in an otherwise all-DNA sequence. The influence of pH, divalent metal cation, hybridization, and secondary structure on the susceptibility of the thio linkage towards transesterification is investigated in an effort to better understand the metal-phosphorothioate interactions and the basis for catalysis. In addition to the chemical cleavage, we have examined the hammerhead ribozyme mediated cleavage of the 5'-bridging phosphorothioate linkage specifically to test the hypothesis that the ribozyme employs a second metal cofactor, which functions as a Lewis acid, to catalyze transesterification. The results of our kinetics experiments do not support this double-metal model. PMID- 9222499 TI - Construction of hairpin ribozymes with a three-way junction. AB - Hairpin ribozymes with high cleavage activities were designed. An extra sequence was introduced at the 3'-end of the hairpin ribozyme to increase the binding to the substrate RNA, as compared to the wild-type hairpin ribozyme. A three-way junction (TWJ) was formed between the newly designed ribozyme and the substrate RNA. The complex with a solid TWJ showed less RNA cleavage activity than the wild type hairpin ribozyme. However, the ribozyme with a TWJ with five unpaired bases or propandiol phosphate linkers had higher cleavage activity than the parent ribozyme without the TWJ. When a cis-cleavage system, in which the 5'-end of the substrate RNA was conjugated to the 3'-end of the ribozyme, was employed, the complex with the TWJ containing unpaired bases was also cleaved faster than the complex with the solid TWJ. This suggested that these differences in the cleavage activities were derived from the confirmation, and this was proven by nondenaturing gel electrophoresis. The TWJ hairpin ribozyme containing unpaired bases is able to bind strongly with substrate RNAs and to cleave them efficiently. Since the three-way ribozyme presented here is more active than the wild-type ribozyme, this type of ribozyme can serve as a more efficient tool to control RNA activities in vitro and in vivo. PMID- 9222500 TI - RNA-RNA interactions between oligonucleotide substrates for aminoacylation. AB - RNA stem-loop microhelices with helix sequences based on tRNA acceptor stems can be charged with specific amino acids. Experiments were designed to test the possibility that microhelices could laterally associate through complementary loop sequences and thereby bring their attached aminoacyl groups close enough together to form a peptide bond. Computer simulations suggested that formation of such complexes would be sensitive to the number of loop nucleotides needed to span the grooves of the quasi-continuous helix of the intermolecular pseudoknot so formed. These predictions were conformed experimentally by observation of complex formation sensitivity to loop size. Complexes with optimized loop sizes had apparent bimolecular dissociation constants of approximately 100 nM with only three complementary base pairs between the respective loops. Single nucleotide substitutions that disrupted the predicted intermolecular loop-loop base-pairing abolished detectable association. Similarly, placing a gap between the short helix formed by loop-loop pairing and the adjacent acceptor stems also diminished complex formation. These experiments establish an experimental basis for microhelix association for peptide synthesis. PMID- 9222501 TI - The selection in vivo and characterization of an RNA recognition motif for spectinomycin. AB - Ribonucleoprotein (RNP) complexes participate in almost all macromolecular processes, including RNA processing, protein synthesis, and the signal recognition of proteins targeted for export. An understanding of these processes requires detailed knowledge of interactions at the molecular level, which has evidently been difficult due to the size and complexity of the particles. Fragmentation of large RNP complexes into functional subdomains is proven to be a successful in vitro strategy to probe ligand interactions at the molecular level. We reasoned that RNA molecules expressed in vivo may fold in such a manner as to mimic a drug binding site present on the intact ribosome. If expressed at sufficient levels, the RNA would sequester the antibiotic thereby permitting the continued function of the ribosome and consequently allow the cell to survive in the presence of the drug. Evidence is presented here in support of this RNA fragment-rescue concept following the selection and characterization of RNA fragments that confer resistance to the antibiotic spectinomycin. PMID- 9222502 TI - Post-SELEX combinatorial optimization of aptamers. AB - In vitro selection techniques provide a means of isolating nucleic acid ligands for binding to particular protein targets. Although most aptamers have quite high affinities for their target proteins, it has been shown that post-SELEX modification can result in further enhancement of binding affinity, as well as other desired properties. This has led to the current development of a more systematic approach to aptamer optimization using a combinatorial screening methodology. PMID- 9222503 TI - In vitro evolution used to define a protein recognition site within a large RNA domain. AB - A minimum of 460 nucleotides of 16S ribosomal RNA are needed to fold the target site for E. coli ribosomal protein S4, although a much smaller region within this large domain is protected from chemical reagents by the protein. Starting with a 531-nucleotide tRNA fragment, cycles of mutagenesis, selection with S4, and amplification ('in vitro evolution') were used to obtain a pool of 30 RNA sequences selected for S4 recognition but approximately 30% different from wild type. Numerous compensatory base pair changes have largely preserved the same secondary structure among these RNAs as found in wild-type sequences. A 20-base deletion and a single nucleotide insertion are among several unusual features found in most of the selected sequences and also prevalent among other prokaryotic rRNAs. Most of the compensatory base changes and selected features are located outside of the region protected by S4 from chemical reagents. It was unexpected that S4 would select for RNA structures throughout such a large domain; the selected features are probably contributing indirectly to S4 recognition by promoting correct tertiary folding of the region actually contacted by S4. The role of S4 may be to stabilize this domain (nearly one-third of the 16S rRNA) in its proper conformation for ribosome function. PMID- 9222505 TI - RNA aptamers that specifically bind to a K Ras-derived farnesylated peptide. AB - RNA aptamers were selected against an affinity column containing a farnesylated peptide modeled after the carboxyl terminus of K ras, the major oncogenic form of this small G protein family. After 10-rounds of selection, 25% of the RNA applied to the column could be specifically eluted. Sequence analysis of the binding RNA aptamers revealed two consensus sequences--GGGUGGG and GGGAGG. Quantitative fluorescence binding studies on two of the high-affinity aptamers, showed a binding affinities of 139 nM and 0.93 microM, respectively for the farnesylated peptide. Binding to the nonfarnesylated peptide was at least 10-fold weaker, showing that the aptamers can recognize the hydrophobic farnesyl moiety. High affinity aptamers could be useful in specifically interfering with oncogenic ras function in particular, and G proteins in general. PMID- 9222504 TI - Interacting RNA species identified by combinatorial selection. AB - RNA molecules were selected from a random sequence library for their ability to bind to an RNA stem-loop target. Oligonucleotides with extensive Watson-Crick complementarity to the RNA ligand were selected against by inclusion of a blocking oligodeoxynucleotide in the binding phase of the selection protocol. After 18 generations of SELEX (systematic evolution of ligands by exponential enrichment) a single RNA family was predominant in the binding population. The winning aptamer RNA bound the target RNA with an apparent Kd = 70 nM. Structural mapping and Fe(II)-EDTA protection indicated that the target RNA interacted with small unpaired loops in the aptamer structure. PMID- 9222506 TI - Inhibition of Rev.RRE complexation by triplex tethered oligonucleotide probes. AB - We have described a class of molecules, called tethered oligonucleotide probes (TOPs), that bind RNA on the basis of both sequence and structure. TOPs consist of two short oligonucleotides joined by a tether whose length and composition may be varied using chemical synthesis. In a triplex TOP, one oligonucleotide recognizes a short single-stranded region in a target RNA through the formation of Watson-Crick base pairs; the other oligonucleotide recognizes a short double stranded region through the formation of Hoogsteen base pairs. Binding of triplex TOPs to an HIV-1 Rev Response Element RNA variant (RREAU) was measured by competition electrophoretic mobility shift analysis. Triplex TOP.RREAU stabilities ranged between -9.6 and -6.1 kcal mol-1 under physiological conditions of pH, salt, and temperature. Although the most stable triplex TOP.RREAU complex contained 12 contiguous U.AU triple helical base pairs, complexes containing only six or nine triple helical base pairs also formed. Triplex TOPs inhibited formation of the RRE.Rev complex with IC50 values that paralleled the dissociation constants of the analogous triplex TOP.RREAU complexes. In contrast to results obtained with TOPs that target two single stranded RRE regions, inhibition of Rev.RREAU complexation by triplex TOPs did not require pre-incubation of RREAU and a TOP: triplex TOPs competed efficiently with Rev for RREAU and inhibited RREAU.Rev complexation at equilibrium. PMID- 9222507 TI - Targeting the Tat-binding site of bovine immunodeficiency virus TAR RNA with a shape-selective rhodium complex. AB - The Tat-binding site of the bovine immunodeficiency virus TAR RNA hairpin has been targeted by Rh(phen)2phi3+ (phen = phenanthroline, phi = 9,10 phenanthrenequinone diimine), a photochemical probe of RNA tertiary structure. The primary site cleaved by the rhodium complex, upon photoactivation, is U24, a base which participates in the novel base triple (with bases A13 and U10) characteristic of this folded RNA. delta-Rh(phen)2phi3+ binds to this site with an affinity of 2 x 10(6) M-1. Upon mutation of U24 and A13 to A24 and U13, respectively, so that the RNA oligomer is unable to form the base triple, site specific cleavage by the rhodium complex is abolished. Moreover, as determined through rhodium photocleavage, at a concentration of 20 microM, Rh(phen)2phi3+ inhibits specific binding of BIV-Tat peptide (2 microM) to its target site. Thus the rhodium complex, in matching its shape to the opened major groove of the properly folded RNA, specifically targets its site and is able to compete for its target with the BIV-Tat peptide. PMID- 9222508 TI - Modulation of nucleic acid structure by ligand binding: induction of a DNA.RNA.DNA hybrid triplex by DAPI intercalation. AB - The aromatic diamidine, DAPI (4',6-diamidino-2-phenylindole), is used as an important biological and cytological tool since it forms highly fluorescent complexes with nucleic acid duplexes via minor groove-directed/intercalative modes of interaction. In this study, we find that DAPI binding can induce the formation of an RNA-DNA hybrid triplex that would not otherwise form. More specifically, through application of a broad range of spectroscopic, viscometric, and molecular modeling techniques, we demonstrate that DAPI intercalation induces the formation of the poly(dT).poly(rA).poly(dT) hybrid triple helix, a structure which does not form in the absence of the ligand. Using UV mixing studies, we demonstrate that, in the presence of DAPI, the poly(rA).poly(dT) duplex and the poly(dT) single strand form a 1:1 complex (a triplex) that does not form in the absence of DAPI. Through temperature-dependent absorbance measurements, we show that the poly(dT).poly(rA).poly(dT) triplex melts via two distinct transitions: initial conversion of the triplex to the duplex state, with the DAPI remaining bound, followed by denaturation of the duplex-DAPI complex to its component single strands and free DAPI. Using optical melting profiles, we show that DAPI binding enhances the thermal stability of the poly(dT).poly(rA).poly(dT) triplex, an observation consistent with the preferential binding of the ligand to the triplex versus the duplex and single-stranded states. Our differential scanning calorimetric measurements reveal melting of the DAPI-saturated poly(dT).poly(rA).poly(dT) triplex to be associated with a lower enthalpy but greater cooperativity than melting of the corresponding DAPI-saturated poly(rA).poly(dT) duplex. Our flow linear dichroism and viscometric data are consistent with an intercalative mode of binding when DAPI interacts with both the poly(dT).poly(rA).poly(dT) triplex and the poly(rA).poly(dT) duplex. Finally, computer modeling studies suggest that a combination of both stacking and electrostatic interactions between the intercalated ligand and the host nucleic acid play important roles in the DAPI-induced stabilization of the poly(dT).poly(rA).poly(dT) triplex. In the aggregate, our results demonstrate that ligand binding can be used to induce the formation of triplex structures that do not form in the absence of the ligand. This triplex-inducing capacity has potentially important implications in the design of novel antisense, antigene, antiviral, and diagnostic strategies. PMID- 9222509 TI - Modulation of the Rev-RRE interaction by aromatic heterocyclic compounds. AB - The HIV-1 Rev protein regulates the nucleocytoplasmic distribution of viral precursor RNAs that encode HIV-1 structural proteins. Rev-mediated viral RNA expression requires a sequence-specific interaction between Rev and a viral RNA sequence, the Rev responsive element (RRE). Because the Rev-RRE interaction is essential for HIV-1 replication, anti-viral agents that selectively block this interaction may be effective anti-HIV-1 therapeutics. Here, we show that certain aromatic heterocyclic compounds, in particular, a tetracationic diphenylfuran, AK.A, can block binding of Rev to its high-affinity viral RNA binding site. AK.A abolishes Rev-RRE interactions at concentrations as low as 0.1 microM. Inhibition appears to be selective and results from competitive binding of the drug to a discrete region within the Rev binding site. Interestingly, the molecular basis for the AK.A-RNA interaction, as well as the mode of RNA binding differs from previously described aminoglycoside Rev inhibitors. Analysis of a variety of aromatic heterocyclic compounds and their derivatives reveals stereo-specific features required for the inhibition. Our results further demonstrate the feasibility of identifying and designing small molecules that selectively block viral RNA-protein interactions. PMID- 9222510 TI - Design and analysis of molecular motifs for specific recognition of RNA. AB - Selective targeting of RNA has become a recent priority in drug design strategies due to the emergence of retroviruses, the need for new antibiotics to counter drug resistance, and our increased awareness of the essential role RNA and RNA structures play in the progression of disease. Most organic compounds known to specifically target RNA are complex, naturally occurring antibiotics that are difficult to synthesize or derivatize and modification of these compounds to optimize interactions with structurally unique RNAs is difficult. The de novo design of synthetically accessible analogues is one possible alternative; however, little is known about the RNA recognition principles on which to design new compounds and limited information on RNA structure in general is available. To contribute to the growing body of knowledge on RNA recognition principles, we have prepared two series of polycationic RNA-binding agents, one with a linear scaffold, the other with a macrocyclic scaffold. We evaluated these compounds for their ability to bind to DNA and RNA, as well as to a specific RNA, the regulatory sequence, RRE, derived from HIV-1, by using thermal melting, circular dichroism, and electrophoresis gel shift methods. Out results suggest that cationic charge centers of high pKa that are displayed along a scaffold of limited flexibility bind preferentially to RNA, most likely within the major groove. Related derivatives that bind more strongly to DNA more closely mimic classical DNA minor-groove binding agents. Several of the macrocyclic polycations expand on a new binding motif where purine bases in duplex RNA are complexed within the macrocyclic cavity, enhancing base-pair opening processes and ultimately destabilizing the RNA duplex. The results in this report should prove a helpful addition to the growing information on molecular motifs that specifically bind to RNA. PMID- 9222511 TI - Discovery of selective, small-molecule inhibitors of RNA complexes--I. The Tat protein/TAR RNA complexes required for HIV-1 transcription. AB - We have developed a therapeutic program focusing on the inhibition of a human immunodeficiency virus-1 specific protein-RNA interaction. This program begins with a search for small organic molecules that would interfere with the binding of Tat protein to TAR RNA. The methodologies chosen to study the HIV-1 Tat-TAR interaction and inhibition include gel mobility shift assays, scintillation proximity assays, filtration assays, and mass spectrometry. These methods helped establish in vitro high-throughput screening assays which rapidly identified Tat TAR inhibitors from our corporate compound library. Tat-activated reporter gene assays were then used to investigate the cellular activities of the Tat-TAR inhibitors. The cellular activity, selectivity, and toxicity data for select Tat TAR inhibitors were determined. Evaluation of both the cellular data and the Tat TAR inhibition results led to further testing in anti-HIV-1 infection assays. PMID- 9222512 TI - Discovery of selective, small-molecule inhibitors of RNA complexes--II. Self splicing group I intron ribozyme. AB - Self-splicing group I intron RNA was chosen as a potential therapeutic target for small-molecule intervention. High-throughput screening methodologies have been developed to identify small organic molecules that regulate the activities of these catalytic introns. Group introns derived from pathogenic Pneumocystis carinii and phage T4 were used as model systems. Inhibitors identified from a library of approximately equal to 150,000 compounds were shown to regulate biochemical reactions including the two-step intron splicing and an RNA ligation catalyzed by the group I introns. These inhibitors provide a unique opportunity to understand small-molecule recognition of the self-splicing RNA. The methodologies developed for group I introns should be applicable to studies of other RNA systems. PMID- 9222513 TI - Selectivity of F8-actinomycin D for RNA:DNA hybrids and its anti-leukemia activity. AB - Although many compounds have been found that bind to DNA in various ways and exhibit various biological activities, few compounds that specifically bind to RNA or RNA:DNA hybrids are known, even though such compounds are expected to have important biological properties. For example, one characteristic function of the retroviruses, which is generally not found in eukaryotic cells, is the production of an RNA:DNA hybrid in the viral replication phase. If an agent is designed to bind only to an RNA:DNA hybrid, and not to DNA or to RNA, such an agent might be able to inhibit specifically the RNase H activity of retroviral reverse transcriptase, and therefore suppress viral replication. Actinomycin D is known to bind to double-stranded DNA, but not to RNA, because steric hindrance between the 2-amino group of the phenoxazone ring and the 2'-hydroxyl group of RNA prevents intercalation of the compound. However, if the > C-H moiety at the 8 position of the phenoxazone ring is replaced by a > C-F, a possible hydrogen-bond acceptor, this analogue (8-fluoro-actinomycin D, F8AMD) might be able to bind intercalatively to an RNA:DNA hybrid by forming an additional hydrogen bond between F8 and the 2'-hydroxyl group of the guanosine ribose. To test this hypothesis, the crystal structure of d(GAAGCTTC)2-F8AMD has been determined at 3.0 A resolution. Based on this crystal structure, a model in which F8AMD binds into the hybrid r(GAAGCUUC):d(GAAGCTTC) has been built using molecular mechanics and dynamic methods. These structural studies indicate that F8AMD binds intercalatively to a B-form double-stranded DNA whereas the drug intercalates into an RNA:DNA hybrid taking an A-form conformation. In the RNA:DNA hybrid complex, the F8 atom is located so as to be able to interact to an O2' hydroxyl group with either an O-H...F hydrogen bond or H+...F- electrostatic interaction. This interaction might stabilize the F8AMD molecule in the RNA:DNA hybrid. A binding study indicates that both actinomycin D (AMD) and F8AMD bind intercalatively not only to double-stranded DNAs, but also to RNA:DNA hybrids. Although the overall binding capacity of F8AMD (k = 4.5 x 10(5) M-1) is reduced slightly in comparison with AMD itself (k = 1.8 x 10(6) M-1), F8AMD tends to bind relatively more favorably than AMD to the RNA:DNA hybrids. The drugs' effects on RNA synthesis in HeLa cells indicates that the binding capacities of AMD and F8AMD correlates strongly to their RNA synthesis inhibitory activities. F8AMD required a concentration of 78 nM to inhibit RNA polymerase activity in HeLa cells by 50%, whereas AMD reached the same inhibitory level at 30 nM. Surprisingly, F8AMD exhibits unique selectivity against leukemia cells as does another C8-derivatized AMD analogue, N8AMD. F8AMD inhibits 50% of leukemia cell growth at less than 1.0 nM whereas 10- to 130-fold-higher drug concentrations are required to inhibit the growth of other tumor cell lines by 50%. The GI50 value of F8AMD for leukemia cells is the lowest among the GI50 values for all other AMD derivatives tested. By contrast, AMD is quite potent and kills most cells at less than 50 nM concentration, but it does not show any selectivity for certain cell lines. This indicates that AMD should have very limited use as an antitumor agent. It is difficult to rationalize why F8AMD and N8AMD show such strong selectivity against leukemia cells. However, this study and our previous study (J. Am. Chem. Soc. 1994, 116, 7971) indicated that F8AMD and N8AMD tended to bind more favorably to RNA:DNA hybrids. Thus, the unique antileukemia selectivity shown by F8AMD and N8AMD might be used by the agents binding to RNA:DNA hybrids rather than to double-stranded DNA. PMID- 9222514 TI - Using guanidinium groups for the recognition of RNA and as catalysts for the hydrolysis of RNA. AB - The guanidinium functional group is commonly used in nature to recognize and bind anions through ion pairing and hydrogen bonding. Specific hydrogen-bonding patterns can be found in crystal structures of simple guanidinium salts. Analysis of these simple salts reveals a variety of features which are found in natural systems. These features have been applied to a series of artificial phosphodiesterases for RNA. These receptors incorporate guanidinium groups positioned to mimic the hydrogen-bonding patterns found in simple guanidinium salts and natural enzymes. This paper outlines general guanidinium hydrogen bonding patterns. Next, the complexation of phosphodiesters with a series of artificial receptors are analyzed in terms of counterions, solvent mixtures, and cavity flexibility. In addition, strategies to enhance catalysis through a pKa analysis of phosphoranes are addressed. Next, we describe how our findings were incorporated into second generation receptors/catalysts. Finally, our future work is discussed. PMID- 9222515 TI - Effect of ribonucleotide substitution on nucleic acid bulge recognition by neocarzinostatin. AB - Bulged RNA structures are not as good substrates for cleavage by the enediyne antibiotic neocarzinostatin chromophore in the general base-catalyzed reaction as are DNA bulges. In an effort to determine why this is so, we have systematically substituted ribonucleotide residues in a DNA bulged structure (CCGATGCG.CGCAGTTCGG) (cleaved residue is underlined) known to be an excellent substrate. It was found that ribonucleotide substitution at the bulge target site, as well as at other regions involving duplex formation had a small effect on the cleavage reaction, unless either of the two strands was entirely of the ribo form. By contrast, changing the A.T base pair on the 5' side of the target nucleotide (T residue) to the ribo A.U resulted in an 87% decrease in cleavage; in fact, conversion of the A alone to the ribo form caused a 68% loss in cleavage. This result can be understood from the recent solution structure of the complex formed between an analogue of the drug radical species and a bulged DNA (Stassinopoulos, A.; Ji, J.; Gao, X.; Goldberg, I.H. Science 1996, 272, 1943), since the 2' hydroxyl group of the ribo A would be expected to clash sterically with the 7"-O-methyl moiety of the drug. Additional studies on substrate bulge dependent drug product formation and protection against spontaneous drug degradation support the cleavage experiments, and imply that bulge-specific drug binding is required for efficient cleavage. PMID- 9222516 TI - Selective cleavages of tRNAPhe with secondary and tertiary structures by enediyne antitumor antibiotics. AB - Some enediyne antitumor antibiotics induce site-selective cleavages for yeast tRNA(Phe) with three-dimensional structure. Of special interest is the fact that tRNA(Phe) is specifically cleaved at the anticodon arm regions by C-1027 and esperamicin A1 in the presence of Mg2+ ions. Although neocarzinostatin strongly breaks tRNA(Phe) at 5'-GPu steps in the absence of magnesium ions, its cleavage ability is completely lost in the presence of 100 microM Mg2+ ions. Dynemicin A, which favors an intercalative binding, causes no strand scissions for the RNA with secondary and tertiary structures. This cutting of tRNA(Phe) may reveal that RNA as well as DNA constitutes a therapeutically relevant target for certain enediyne antitumor antibiotics. PMID- 9222517 TI - On the chemistry of RNA degradation by Fe.bleomycin. AB - The chemistry of RNA degradation by Fe.bleomycin was studied using two RNA substrates that are modified efficiently at a small number of sites by the antitumor antibiotic. Cleavage of tRNAHis precursor transcript by Fe(II).BLM A2 was shown to require O2; cleavage was also observed when the same substrate was treated with Fe(III).BLM A2 + H2O2. Consistent with earlier observations made for DNA, the extent of tRNAHis precursor cleavage was greater for Fe(II).BLM A5 than for Fe(II).BLM A2; the least cleavage was obtained using Fe(II).BLM demethyl A2. By the use of 32P end labeled tRNAHis precursor transcript that was also 3H labeled within the uracil moieties, it was shown that release of uracil was nearly stoichiometric with tRNA strand scission by Fe(II).BLM A2. Nonetheless, treatment of the tRNAHis with hydrazine following BLM-mediated cleavage indicated formation of a new product that must have derived from a BLM-induced lesion. Also employed for characterization of BLM cleavage of RNA were the octanucleotides CGCTAGCG, C3-ribo-CGCTAGCG and C3-ara-CGCTAGCG. Analysis of the products of cleavage indicates that Fe.BLM is capable of mediating cleavage by abstraction of a H atom either from C-4' H or c-1' H of the chimeric oligonucleotides. PMID- 9222518 TI - Subcortical aphasia. AB - We critically review the literature on subcortical aphasia, suggest that a number of traditional concepts regarding mechanisms of aphasia are inconsistent with now abundant data, and propose several new hypotheses. The absence of aphasia in 17 reported cases of dominant hemisphere striatocapsular infarction and the finding of nearly every conceivable pattern of language impairment in 33 different reported cases of striatocapsular infarction provide strong evidence against a major direct role of the basal ganglia in language and against disconnection or diaschisis as mechanisms of nonthalamic subcortical aphasia. However, detailed consideration of the vascular events leading to striatocapsular infarction strongly suggests that associated linguistic deficits are predominantly related to sustained cortical hypoperfusion and infarction not visible on structural imaging studies. Thalamic disconnection, as may occur with striatocapsular infarcts with extension to the temporal stem and putamenal hemorrhages, may also contribute to the language deficits in some patients. Review of the literature on thalamic infarction, in conjunction with previously unreported anatomic details of four cases, suggests that what infarcts in the tuberothalamic artery territory and the occasional infarcts in the paramedian artery territory associated with aphasia have in common is damage to the frontal lobe-inferior thalamic peduncle nucleus reticularis-center median system that may be involved in regulating the thalamic gate in attentional processes. Disruption of attentional gating in the pulvinar and lateral posterior nuclei resulting from such lesions may impair selection of specific neuronal networks in the projection field of these nuclei that serve as the substrate for lexical-semantic function, which is in effect a disruption of a type of working memory, as defined by Goldman-Rakic. We define this as a defect of selective engagement. PMID- 9222519 TI - Mechanisms of and misconceptions about subcortical aphasia. PMID- 9222520 TI - Subcortical aphasia(s): a controversial and promising topic. AB - Many aspects of subcortical aphasias are not accounted for by a unique pathophysiological mechanism; rather the diversity of the observed symptoms suggests that many functional ensembles, more or less related, to language functions may be disturbed by subcortical lesions. However, such disorders are only moderate in intensity and subcortical aphasias certainly constitute an interesting model for studying the dynamics of recovery of language functions and related neural systems, combining neurolinguistic concepts with advanced functional imaging techniques. PMID- 9222522 TI - Subcortical aphasia: still a useful concept? PMID- 9222523 TI - Subcortical aphasia and the problem of attributing functional responsibility to parts of distributed brain processes. AB - N&C's discussion is, in places, an exemplar of the sort of rigor and attention to detail that will bring us closer to an understanding of the functional organization of the brain. Indeed, it is this level of work that pushes us to reflect on the assumptions that undergird our research efforts. Our criticisms have developed four main points. First, the level of rigor applied to the consideration of basal ganglionic aphasia should extend to each application of the CPC method (thalamic aphasia included). Second, in our haste to identify specific brain systems with distinct cognitive functions we should not neglect the more basic question of the causal mechanisms by which the brain organizes behavior. Questions of "direct" versus "indirect" involvement of a particular organ in a cognitive function are only likely to distract our attention from this more basic and less inferentially perilous issue. Third, pure cases should no longer be considered touchstones against which all behavioral disturbances are measured. Reifying such ideals is more likely to shroud than reveal the brain's true complexity. Finally, the functions that we enshrine in particular brain regions should explain the particular character of the symptoms observed when they are damaged and should admit of independent verification. PMID- 9222524 TI - Nitric oxide inhibits neuronal activity in the supraoptic nucleus of the rat hypothalamic slices. AB - The presence of abundant nitric oxide synthase (NOS) in magnocellular neurons of the rat hypothalamus suggests that nitric oxide (NO) may be involved in controlling the release of oxytocin and vasopressin. To test this possibility, we examined the effect of NO-related drugs on extracellular discharges of 124 supraoptic nucleus (SON) neurons from slices of rat hypothalamus in vitro. Twenty three (43%) of 53 neurons were inhibited by sodium nitroprusside (SNP), a spontaneous releaser of NO, at 1-3 mM. This inhibition was prevented by preincubation of the slices with 1 microM hemoglobin, an inactivator of NO (n = 14), whereas hemoglobin alone enhanced neuronal activity in seven (35%) of 20 neurons. L-Arginine (1 mM), a precursor of NO, inhibited neuronal activity in five (36%) of 14 neurons, while D-arginine (1 mM), the inactive counterpart of L arginine, was ineffective (n = 12). N-omega-nitro-L-arginine methyl ester (L NAME, 10 microM), an inhibitor of NOS, also enhanced neuronal activity in five (29%) of 17 neurons, while N-omega-nitro-D-arginine methyl ester (DNAME, 10 microM), the inactive enantiomer of L-NAME, was without effect (n = 11). Together, our data show that NO exerts predominantly an inhibitory effect on SON neurons and may serve as a negative feedback loop in controlling release of oxytocin and vasopressin. PMID- 9222525 TI - Overlapping ipsilateral and contralateral retinal projections to the lateral geniculate nucleus and superior colliculus in the cat: a retrograde triple labelling study. AB - To analyze the relative proportion and distribution of retinal ganglion cells projecting ipsilaterally and contralaterally in the cat, large injections of the fluorescent tracers Fluoro Gold, Fast Blue, and Diamidino Yellow were made in the main layers of the lateral geniculate nucleus (LGN) and superior colliculus (SC). One tracer was injected in both the LGN and SC on one side, and the other two tracers were injected contralaterally, in the LGN and SC, respectively; labelled ganglion cells were charted on retinal whole mounts. Ganglion cells labelled from the LGN and SC were highly intermingled in both the ipsilateral and contralateral retinae. The adopted combinations of tracers allowed the detection of cells double labelled from the SC and LGN, supporting the occurrence of branched retino thalamic axons to the SC. About one-fourth of the ganglion cells labelled from the LGN and SC was located in the eye ipsilateral to the injection. Retrograde labelling from the ipsilateral side was almost entirely confined to the temporal hemiretina. In the contralateral eye, labelled cells were mainly concentrated in the nasal hemiretina, but more than 10% were also detected in the temporal half of the retina. In the latter area, cells displaying the entire range of sizes of the retinal ganglion cells, labelled from the contralateral LGN and SC, were found throughout the entire hemiretina. However, more than 50% of such "wrong" projecting cells were grouped in a strip of 2 mm closest to the nasotemporal division. Control experiments, in which the tracers injections were restricted to the rostral and dorsal portions of the LGN to avoid optic tract contamination, consistently confirmed the occurrence and distribution of the "wrong" projecting cells in the temporal hemiretina. Thus, these latter cells are not grouped in a central strip, where ganglion cells would have the same chance of projecting to the same or to the opposite side, and sparsely distributed in the temporal periphery, as previously believed. Instead, the present findings indicate that the retinal ganglion cells of origin of contralateral projections are distributed more in a continuum, with a naso-temporal gradient of density, across the temporal hemiretina. PMID- 9222526 TI - A three-dimensional multimodality brain map of the nemestrina monkey. AB - A three-dimensional multimodality computerized map of the nemestrina monkey brain was created with serial sectioning and digital imaging techniques. An adult female Macaca nemestrina (pigtail macaque) weighing 7.2 kg was used in constructing this atlas. CT, PET, and MRI were performed on the monkey before the specimen's head was frozen and cryoplaned. Closely spaced (50 microns) images of the specimen blockface were then digitally acquired and modified to produce whole head and brain-only 3D image sets. The resulting data sets were organized into a digital volume and repositioned into a stereotaxic coordinate system defined by Horsley and Clark in 1908 [7]. Orthogonal images were obtained by digitally resampling the volume in order to produce a full set of coronal, sagittal, and horizontal images. Stereotaxic reference grids were applied to each image indicating the A/P, M/L, or Ho position within the digital volume. Specific anatomic structures were outlined from the cryosection data set and 3D surface models reconstructed. Structural labels indicating nuclei, tracts, and other neuroanatomical features were incorporated into coronally sliced cryosection images spaced at 500 microns. The CT, PET, and MRI data sets were reconstructed into a digital volume and coregistered to the cryosection volume. All images constructed from this 3D map are available for public access via the internet using an anonymous file transfer protocol (FTP) and the World Wide Web (http:@www.loni.ucla.edu). The foremost advantage of this digital map is an integrated multimodality three-dimensional representation of the Macaca nemestrina brain, which is not possible with traditional atlases. PMID- 9222527 TI - Oxidative enzyme activity and soma size in motoneurons innervating the rat slow twitch and fast-twitch muscles after chronic activity. AB - The effects of chronic activity induced by running training on the activity of the mitochondrial enzyme succinate dehydrogenase (SDH) and soma size in motoneurons innervating the slow-twitch soleus (SOL) and fast-twitch extensor digitorum longus (EDL) muscles were studied in rats using the retrograde neuronal tracer Nuclear Yellow. Rats were assigned to control and trained groups that were subjected to treadmill running for 10 weeks (2 h/day, 30 m/min, 5 days/week). After training, both SOL and EDL muscles showed clear adaptations (citrate synthase activity in the SOL muscle, and the fast-twitch oxidative-glycolytic fiber area of the EDL muscle increased significantly after training). The SDH activity of the motoneurons innervating both SOL and EDL muscles was unchanged by training. However, SOL motoneurons of trained rats had a significantly larger soma size and a significantly higher total SDH activity [SDH activity x soma size) than those of control. Total SDH activity was calculated to examine the absolute SDH protein content of the motoneurons. On the other hand, there was no difference in both soma size and total SDH activity of EDL motoneurons between the two groups. These data demonstrate that chronic activity has a considerably stronger impact on soma size and total oxidative enzyme activity of motoneurons innervating slow-twitch rather than fast-twitch muscles. PMID- 9222528 TI - Interferon-gamma alters nerve-induced redistribution of acetylcholine receptors in cultured rat skeletal muscle cells. AB - The influence of recombinant interferon-gamma (rIFN-gamma) on the development of acetylcholine receptor (AChR) aggregates in cocultures of rat embryonic muscle cells and spinal cord neurons was studied by counting the number of AChR aggregates in relation to cholinergic nerve fibers coming to the muscle fibers. rIFN-gamma caused no decrease in the number of cholinergic nerve fibers, but inhibited the increase in the number of AChR aggregates that occurs early during cocultivation and is an early sign in the development of neuromuscular junctions. rIFN-gamma stimulated release of nitric oxide, but no effects on aggregation of AChRs occurred after exposure to a nitric oxide synthase inhibitor, L-NG monomethylarginine, or by the addition of nitroprusside, a generator of nitric oxide. No effect was seen on the number of AChR aggregates when the cultures were exposed to rIFN-gamma at later time points of cocultivation, when the increase in number of AChRs had already occurred. These studies indicate that the key immunoregulatory cytokine IFN-gamma can cause alterations in the early process of synapse formation and that these effects are independent of the nitric oxide release caused by the cytokine. PMID- 9222529 TI - Lack of relationship between thalamic oscillations and attention in rats: differential modulation by an alpha-2 antagonist. AB - A five-choice serial reaction time (5-CSRT) task was used to assess attention in rats. In this behavioral paradigm, the rats are required to spatially discriminate a short visual stimulus that will occur randomly in one of five locations while maintaining a sufficient activity level. The ability of a rat to maintain attention on the task can be measured by counting the choice accuracy (percent correct responses), whereas the probability of premature responses indicates the level of impulsivity. According to previous results [24], rats performing poorly in the task have a lower choice accuracy and make more premature responses than normally behaving individuals, i.e., a clear, inverse correlation was observed between choice accuracy and impulsiveness of rats. Methylphenidate, a psychostimulant that has been shown to alleviate the symptoms in attention deficit-hyperactivity disorder (ADHD), improved the choice accuracy of poor performing rats in this task [24]. The present results show that the correlation between choice accuracy and impulsivity exists also when the rats are tested using a reduced stimulus intensity or curtailed stimulus duration. The results of a pharmacological experiment suggested that atipamezole (30, 300, or 1000 micrograms/kg), a potent and specific alpha-2 antagonist that is known to increase the activity of monoaminergic systems in the brain, did not affect the percent correct responses in poor performers or in controls tested either at the baseline conditions or at a curtailed stimulus duration (which impaired their choice accuracy). At the doses of 300 and 1000 micrograms/kg, however, atipamezole slightly increased the probability of premature responses in all group of rats. The results of an electrophysiological study indicated that the poor choice accuracy or impulsiveness of rats is not related to the amount of cortically recorded spike-wave discharges/high voltage spindle (HVS) activity, which reflect thalamo-cortical oscillation. Atipamezole dose-dependently reduced the incidence and duration of HVSs. The present data, therefore, indicate that (a) alpha-2 antagonist treatment is not superior to methylphenidate treatment when investigated using acute administrations of the agents in poor performers of the 5-CSRT task, and (b) thalamic oscillations are not the reason for the attention deficit of rats in this model of ADHD. The relationship between choice accuracy and impulsivity is discussed. PMID- 9222530 TI - Maternal exposure to delta 9-tetrahydrocannabinol (delta 9-THC) alters indolamine levels and turnover in adult male and female rat brain regions. AB - Perinatal exposure to delta 9-THC has been shown to produce effects on brain development. In this study we evaluated the changes induced by maternal exposure to delta 9-THC (5 mg/kg per day) from gestational day 5 to postnatal day 24 in eight discrete brain areas on the central serotoninergic system in both adult male and female rats. These result show that maternal exposure to delta 9-THC from gestational day 5 to postnatal day 24 affects development of the various central indoleaminergic system of the offsprings brain. Perinatal exposure to delta 9-THC decreased the levels of 5-HT in hypothalamus and rostral neostriatum in exposed males, and also decreased the levels of 5-HT in ventral hippocampus, septum, and midbrain raphe nuclei in both exposed males and females. Perinatal exposure to delta 9-THC increased the levels of 5-HIAA in dorsal hippocampus, hypothalamus, septum, midbrain raphe nuclei, and rostral neostriatum in exposed males and females. We have also found differences between nonexposed males and females in several brain regions. Our results confirm a regional and sexual specificity in endogenous levels of indoleamine after perinatal delta 9-THC treatment, being the midbrain raphe nuclei the most affected area. PMID- 9222532 TI - Anticonvulsant activity of new and potent inhibitors of nitric oxide synthase. AB - The effects of new and potent NOS inhibitors, S-methyl-L-thiocitrulline (S-Me TC), 3-bromo 7-nitro indazole (3-Br-7-NI), and 1-(2-trifluoromethylphenyl) imidazole (TRIM), were examined on the pilocarpine-induced seizures in mice. 3-Br 7-NI and TRIM decreased the frequency of status epilepticus and mortality, while TRIM. In addition, significantly reduced the incidence of seizures. The latencies to onsets of seizures, status epilepticus, and mortality were significantly prolonged by all three NOS inhibitors, while duration of seizures was reduced by 3-Br-7-NI and TRIM. These data suggest an excitatory effect of NO in the neuronal structure involved in the pilocarpine-induced seizures. PMID- 9222531 TI - Chlorpyrifos interferes with cell development in rat brain regions. AB - Chlorpyrifos, one of the most widely used pesticides, exhibits greater toxicity during development than in adulthood. We administered chlorpyrifos to neonatal rats in doses spanning the threshold for systemic toxicity and examined developing brain regions (brainstem, forebrain, cerebellum) for signs of interference with cell development using markers for cell packing density and cell number (DNA concentration and content) and cell size (protein/DNA ratio). Neonatal rats given 5 mg/kg of chlorpyrifos on postnatal days 1-4 showed significant mortality and the survivors exhibited severe cell loss in the brainstem; brainstem growth was maintained by enlargement of the remaining cells. This effect was not seen at 1 mg/kg, a dose that did not compromise survival or growth, nor was there any adverse effect at either dose in the forebrain, despite the fact that both brainstem and forebrain possess comparable cholinergic projections. When chlorpyrifos was administered later, on days 11-14, the major target for cell loss shifted from the brainstem to the forebrain and in this case, effects were seen at doses that did not compromise survival or growth. The loss of forebrain cell number occurred between 15 and 20 days of age rather than during the chlorpyrifos treatment. The cerebellum differed from the other regions in that it showed short-term elevations of DNA after chlorpyrifos exposure in either early or late postnatal periods; nevertheless, values then regressed to subnormal in parallel with the loss of cells in other regions. Thus, chlorpyrifos likely causes delayed cell death. Although regions rich in cholinergic projections, such as brainstem and forebrain, may be more affected than noncholinergic regions (cerebellum), the maturational timetable of each region (brainstem earliest, forebrain intermediate, cerebellum last) appears to be more important in setting the window of vulnerability. These results indicate that, even when growth or survival are unaffected, chlorpyrifos produces cellular deficits in the developing brain that could contribute to behavioral abnormalities. PMID- 9222533 TI - The distribution of p75 neurotrophin receptor-immunoreactive cells in the forebrain of the common marmoset (Callithrix jacchus). AB - The distribution of neurones that could be stained immunohistochemically with antibody to the p75 neurotrophin protein was studied in the forebrain of the common marmoset. The p75-immunoreactive forebrain cells appear to correspond to choline acetyltransferase-immunoreactive (i.e., cholinergic) neurones. Two populations of cells could be distinguished on the basis of the intensity of p75 immunostaining. Moderately stained cells correspond to cholinergic interneurones of the caudate and putamen, while intensely stained cells correspond to the cholinergic neurones projecting to the cortex, amygdala, and hippocampus, located in the septum, diagonal band, and basal nucleus of Meynert. The distribution of cells of the diagonal band/basal nucleus complex is more extensive in the marmoset than in other primate species, extending into parts of the postcommissural fornix via the posterior septum, and by small projections dorsal to the anterior commissure and via the thalamic fasciculus from the basal nucleus; the posterior extent of the basal nucleus continues extensively into the lamina between the globus pallidus and the putamen. PMID- 9222534 TI - Ventromedial nuclei of the hypothalamus are involved in the phase advance of temperature and activity rhythms in food-restricted rats fed during daytime. AB - Daily rhythms are synchronized to the light-dark cycle (LD) via a circadian clock located in the suprachiasmatic nuclei. A timed caloric restriction phase advances daily rhythms of body temperature and wheel-running activity in rats kept under LD. Because lesions of the ventromedial hypothalamic nuclei (VMH) prevent the fasting-induced changes in the day-night pattern of activity, it was hypothesized that the VMH might participate in the caloric restriction-induced phase changes. To test this hypothesis, rats with electrolytic or ibotenic acid lesions of VMH and control rats were fed 2 h after lights on 50% of ad lib food intake. During the preceding fed state, rats with electrolytic lesions of VMH displayed a less marked day-night difference in locomotor activity and a phase-advanced acrophase of temperature rhythm (2 h) compared to those of sham-operated rats. These effects were not found in fed rats with ibotenic lesions of VMH, suggesting that these effects of electrolytic lesions were due to disruption of undetermined fibers of passage. In response to a timed caloric restriction, the nocturnal peak of temperature rhythm was phase advanced by 7 h in sham-operated rats. Their day night pattern of activity was also phase advanced towards the time of feeding. In both groups of food-restricted VMH-lesioned rats, the acrophase of temperature rhythm plateaued 3 h later than in sham-operated group. The phase advance of body temperature was, therefore, reduced to 4 h by ibotenic lesions of VMH and to 2 h by electrolytic lesions. Except for a feeding-associated component of activity expressed in control and VMH-lesioned rats, no significant change in day-night pattern of activity was detected in VMH-lesioned rats, either by electrolytic or ibotenic lesions. These results indicate that neuronal damage of the VMH limits the phase-advancing properties of a timed caloric restriction on the daily rhythms of temperature and locomotor activity. PMID- 9222535 TI - Regulation of prepulse inhibition by ventral pallidal projections. AB - The acoustic startle reflex is inhibited by the presentation of a weak auditory prestimulus 30-500 ma prior to the starting stimulus. Previous studies have demonstrated that prepulse inhibition (PPI) of acoustic startle is regulated by GABAergic activity in the ventral pallidum. Ventral pallidal efferents include major projections to the pedunculopontine tegmental nucleus (PPTg), subthalamic nucleus (STN), and mediodorsal thalamus (MD). We used lesion and intracerebral infusion techniques to determine the relevance of these projections to the ventral pallidal regulation of PPI. Consistent with previous results, PPTg lesions significantly reduced PPI in all startle sessions, while MD lesions significantly reduced PPI only under certain experimental conditions. STN lesions failed to alter PPI, but they did significantly disrupt amphetamine-induced locomotion, verifying the behavioral effectiveness of these lesions. Infusion of the GABA-A agonist muscimol into either the PPTg or the MD significantly reduced PPI. Ventral pallidal projections to the PPTg and to the MD thus appear to regulate PPI, possibly via a GABAergic mechanism. Pallidal projections to the STN may regulate other behavioral processes such as locomotor activity, but they do not appear to regulate sensorimotor gating of the acoustic startle reflex. PMID- 9222536 TI - Nerve growth factor, central nervous system apoptosis, and adrenocortical activity in aged Fischer-344/brown Norway F1 hybrid rats. AB - During aging there is a progressive loss of neuronal function in the basal forebrain that results in cognitive impairment and cholinergic deficits. While altered neurotrophin (NT)-mediated signal transduction may account for some age associated deficits, there are differences in the extent of NT responsiveness among different laboratory rat strains. Here we measured nerve growth factor (NGF) protein levels and fragmented DNA in the CNS, and basal and NGF-stimulated activity levels of the hypothalamus-pituitary-adrenocortical axis (HPAA) in 3-, 18-, and 30-month-old Fischer-344/Brown Norway rats. Our results show that while there is no age-associated differences in NGF protein levels, in aged Fischer 344/Brown Norway rats, there are increases in levels of immunoreactive fragmented DNA in the CNS and in adrenocortical responses to the peripheral administration of NGF. These data contribute to the characterization of the Fischer-344/Brown Norway F1 hybrid rat and provide baseline values useful for future studies on aged CNS. PMID- 9222537 TI - Distribution and time course of appearance of "dark" neurons and EEG activity after amygdaloid kainate lesion. AB - To determine the extent and time course of local and distant neuronal damage produced by microiontophoretic administration of kainic acid (KA) into the central amygdaloid nucleus, distribution of neuronal damage was compared in various brain areas after different survival times. For demonstration of damaged, so-called "dark" neurons, a newly developed silver stain was employed. In addition, silver staining method was used to visualize microglia cells. In a separate experiment, electroencephalographic (EEG) activity was recorded from the amygdaloid body, hippocampus, and the frontal cortex before and after microiontophoretic KA lesion of the central amygdaloid nucleus. It was observed that (1) even a minute amount of KA into this nucleus caused transient neuronal damage in distant brain areas; (2) the hippocampal formation, subiculum, entorhinal cortex, piriform cortex, and lateral septum were consistently affected; (3) the extent and time course of neuronal damage and appearance of microglia cells varied from area to area; (4) the KA neurotoxicity in distant brain areas appeared to depend on specific excitatory circuits, especially in the hippocampal formation; (5) the appearance and time course of pathologic EEG activity paralleled the appearance of dark neurons; and (6) the absence of pathologic EEG activity and the lack of massive neuronal loss or microglia proliferation in distant brain areas of rats surviving longer than 48 h suggested that these areas may have recovered both morphologically and functionally. Although details of cellular mechanism responsible for development of "dark" degeneration of neurons are not known, the silver method employed in the present study proved to be sensitive, useful tool for fine histological analyses of early and distant consequences of excitotoxic lesions. PMID- 9222538 TI - Sixteenth Gaddum Memorial Lecture December 1996. Neuroimmune interactions: the role of cytokines. PMID- 9222539 TI - Inhibition by heterologously-expressed P2Y2 nucleotide receptors of N-type calcium currents in rat sympathetic neurones. AB - The P2Y2 nucleotide receptor has previously been shown to stimulate phosphoinositide breakdown. We now show that, when P2Y2 receptors are heterologously expressed by cRNA injection into dissociated rat sympathetic neurones, activation of these receptors by uridine 5'-triphosphate (UTP) or adenosine 5'-triphosphate (ATP) inhibits the N-type voltage-gated calcium current by approximately 65%, with an IC50 of 0.5 microM. Thus, the same molecular species of nucleotide receptor can link to two different effector pathways. PMID- 9222540 TI - Inhibition of 5-hydroxytryptamine-induced phosphoinositide hydrolysis and Ca2+ mobilization in canine cultured tracheal smooth muscle cells by phorbol ester. AB - 1. Regulation of the increase in inositol-1,4,5-trisphosphate (IP3) production and intracellular Ca2+ concentration ([Ca2+]i by protein kinase C (PKC) was investigated in canine cultured tracheal smooth muscle cells (TSMCs). Stimulation of TSMCs by 5-hydroxytryptamine (5-HT) caused an initial transient [Ca2+]i peak followed by a sustained elevation of [Ca2+]i in a concentration-dependent manner. 2. Pretreatment of TSMCs with phorbol 12-myristate 13-acetate (PMA, 1 microM) for 30 min blocked the 5-HT-induced IP3 formation and Ca2+ mobilization. This inhibition was reduced after the cells had been incubated with PMA for 8 h, and within 48 h the 5-HT-induced Ca2+ mobilization reached the same extent as control cells. 3. The concentration of PMA that gave half-maximal inhibition of 5-HT induced increase in [Ca2+]i was 4 nM. Pretreatment of TSMCs with staurosporine (1 microM) of GF109203X (0.1 microM), PKC inhibitors, inhibited the ability of PMA to attenuate 5-HT-induced responses, suggesting that the inhibitory effect of PMA was mediated through the activation of PKC. 4. In parallel with the effect of PMA on 5-HT-induced IP3 formation and Ca2+ mobilization, the translocation and down regulation of PKC isozymes were determined by Western blot analysis in TSMCs. Analysis of cell extracts by Western blotting with antibodies against different PKC isozymes revealed that TSMCs expressed PKC-alpha, beta I, beta II, delta, epsilon, theta and zeta. With PMA treatment of the cells for various times, translocation of PKC-alpha, beta I, beta II, delta, epsilon, and theta from the cytosol to the membrane was seen after 5 min, 30 min, 2 h, and 4 h treatment. However, 24 h treatment caused a partial down-regulation of these PKC isozymes PKC-zeta was not significantly translocated and down-regulated at any of the times tested. 5. In conclusion, these results suggest that activation of PKC may inhibit the receptor-mediated phosphoinositide hydrolysis and consequently attenuate the [Ca2+]i increase or inhibit both responses independently. The translocation of PKC-alpha, beta I, beta II, delta, epsilon, and theta induced by PMA caused an attenuation of 5-HT-stimulated IP3 accumulation and Ca2+ mobilization in TSMCs. PMID- 9222541 TI - Renal alpha 2a/d-adrenoceptor subtype function: Wistar as compared to spontaneously hypertensive rats. AB - 1. The alpha 2a/d-adrenoceptor subtype in the rat kidney modulates solute excretion (osmolar clearance). Since the kidney plays a role in chronic regulation of blood pressure, altered renal function may be implicated in the development of hypertension. A second alteration-that of the alpha 2a/d adrenoceptor subtype gene-has also been correlated with hypertension in rats and man. 2. We hypothesized that as a consequence of the altered alpha 2a/d adrenoceptor subtype gene previously shown in spontaneously hypertensive (SH) rats, the increase in osmolar clearance following stimulation of the renal alpha 2a/d-subtype would be attenuated in SH rats as compared to normotensive Wistar rats. In contrast, based on the theory that such functional unresponsiveness of the alpha 2a/d-subtype would be genetically determined, we further hypothesized that in one kidney-one clip (1K-1C) rats, the response to stimulation of the renal alpha 2a/d-subtype would be intact as compared to the normotensive Wistar 1K-sham rats. 3. Male rats were unilaterally nephrectomized under ether anaesthesia. In the 1K-1C rats, a silver clip (diameter 0.254 mm) was also placed around the left renal artery. On the experimental day, rats were administered pentobarbitone (50.0 mg kg-1, i.p.). The carotid artery and jugular vein were cannulated for blood pressure monitoring and saline infusion. The ureter was catheterized for urine collection. A 31 gauge needle was advanced into the renal artery for infusion of the alpha 2a/d-selective agonist, guanfacine (vehicle, 1.0, 3.0 and 10.0 nmol kg-1 min-1 in Wistar and SH rats; vehicle and 10.0 nmol kg 1 min-1 in Wistar 1K-sham and 1K-1C rats). 4. In Wistar rats, guanfacine dose dependently increased urine flow and sodium excretion. An increase in osmolar clearance but not free water clearance was also observed. However, in SH rats guanfacine failed to alter urine flow, sodium excretion, osmolar and free water clearance. In contrast, in both Wistar 1K-sham and 1K-1C rats, guanfacine increased urine flow rate. Again, this response was due solely to an increase in osmolar clearance. At these doses, guanfacine did not alter blood pressure or creatinine clearance during the experiment. 5. In summary, the ability of the alpha 2a/d-adrenoceptor subtype to mediate an increase in osmolar clearance was absent in a genetic model of hypertension, the SH rats. This effect was intact in an acquired model of hypertension (1K-1C rats). This suggested a defective modulation of solute excretion in SH rats which was probably due to alteration of the alpha 2a/d-subtype gene and not secondary to the elevated blood pressure. The altered alpha 2a/d-subtype gene and function may therefore play a causal role in the pathogenesis of hypertension. PMID- 9222542 TI - Hydrogen peroxide induced responses of cat tracheal smooth muscle cells. AB - 1. The effects of hydrogen peroxide (H2O2) (10(-6)-10(-3) M) on membrane potential, membrane currents, intracellular calcium concentration, resting muscle tone and contractions elicited by electrical field stimulation (EFS) and carbachol were examined in cat tracheal strips and isolated smooth muscle cells. 2. H2O2 (10(-4) and 10(-5) M) enhanced the amplitude of contractions and excitatory junction potentials (e.j.p.) evoked by EFS without changing muscle tone and resting membrane potential of the tracheal smooth muscle, and enhanced the contraction induced by carbachol (10(-3) M). At an increased concentration (10(-3) M), H2O2 elevated resting muscle tone and marginally hyperpolarized the membrane in the majority of the cells. 3. In 51 out of 56 cells examined, H2O2 (10(-6)-10(-3) M) elicited an outward current at a holding potential of -40 mV and enhanced the frequency of the spontaneous transient outward current (STOC). In 20 cells the outward current was preceded by a small inward current. In the other cells, H2O2 elicited only an inward current or did not affect the background current. 4. In Ca2+ free solution the action of H2O2 on the resting muscle tone, STOCs, background current and on the current induced by ramp depolarization was significantly reduced. 5. H2O2 (10(-4) M) increased the intracellular ionized calcium concentration both in the absence and presence of external Ca2+. However, the effect developed faster and was of a higher amplitude in the presence of external Ca2+. 6. These results suggest that H2O2 increases intracellular Ca2+, with a subsequent augmentation of stimulation-evoked contractions, and enhances Ca2+ and voltage-sensitive potassium conductance. PMID- 9222543 TI - Combined administration of 5-HT2 and thromboxane A2 antagonists: effects on platelet aggregation and isolated cardiac muscle. AB - 1. To investigate possible mechanisms underlying the ability of combined administration of a 5-hydroxytryptamine2 (5-HT2) antagonist and a thromboxane A2 antagonist to reduce reperfusion-induced arrhythmias, the effects of these drugs alone and in combination on platelet aggregation and on cardiac muscle were determined. 2. Platelet aggregation was measured in whole blood obtained from anaesthetized rats. Concentrations of 5-HT (10 microM) and the thromboxane A2 mimetic U46619 (1 microM) which did not cause aggregation themselves, enhanced the responses to ADP (0.1 microM) and to collagen (1 microgram ml-1). For example, the response of 1.0 +/- 0.5 omega to ADP alone was increased significantly to 6.4 +/- 1.0 omega by 5-HT, 15.5 +/- 2.8 omega by U46619, and 17.3 +/- 1.3 omega when U46619, 5-HT and ADP were added together. 3. In further experiments blood was obtained from rats which had received either the 5-HT2 antagonist, ICI 170,809 (1 mg kg-1), or the thromboxane A2 antagonist. ICI 192,605 (1 mg kg-1 min-1), or both in combination. When ADP was used as the primary aggregating agent, the ability of U46619 alone, or together with 5-HT, to enhance responses was reduced significantly by ICI 192,605 alone and in combination with ICI 170,809. Similar results were obtained with lower doses of ICI 170,809 (0.3 mg kg-1) and ICI 192,605 (0.3 mg kg-1 min-1). 4. When collagen was used as the primary aggregating agent ICI 170,809 (1 mg kg-1) reduced the response to 5-HT (5.0 +/- 0.8 omega versus 10.9 +/- 1.2 omega in controls), and ICI 192,605 (1 mg kg-1 min-1) reduced the response to U46619 (6.8 +/- 2.5 omega versus 11.2 +/- 2.2 omega in control). The greatest reduction of platelet aggregation was seen in blood from rats which had received both antagonists, with the response to U46619 plus 5-HT plus collagen being 2.7 +/- 0.6 omega compared to 14.2 +/- 1.7 omega in controls. In contrast, there was no significant attenuation of platelet aggregation in blood from rats which had received the lower doses of each antagonist alone. Only the combination of ICI 170,809 (0.3 mg kg-1) and ICI 192,605 (0.3 mg kg-1 min-1) reduced the response to U46619 plus 5 HT plus collagen (7.6 +/- 1.4 omega versus 15.0 +/- 0.5 omega in controls). 5. In rat isolated ventricular muscle preparations, ICI 170,809 increased the effective refractory period; e.g. from 39 +/- 4 to 86 +/- 18 ms, 10 min after adding 30 microM to left papillary muscles. ICI 192,605 did not increase the effective refractory period itself and did not alter the ability of ICI 170,809 to prolong the effective refractory period. In the presence of 100 microM ICI 192,605, ICI 170,809 (30 microM) increased the effective refractory period from 38 +/- 7 to 100 +/- 30 ms. 6. These results indicate that the previously observed antiarrhythmic activity of combined administration of the higher doses of ICI 170,809 and ICI 192,605 is unlikely to be due to direct effects on cardiac muscle but could be a consequence of reduced platelet aggregation. PMID- 9222544 TI - Role of lipocortin-1 in the anti-hyperalgesic actions of dexamethasone. AB - 1. The effect of dexamethasone, lipocorton-1(2-26) and an antiserum to lipocortin 1(2-26) (LCPS1) upon the hyperalgesic activities in rats of carrageenin, bradykinin, tumour necrosis factor alpha (TNF alpha), interleukin-1(2), interleukin-6 (IL-6), interleukin-8 (IL-8), prostaglandin E beta (PGE2) and dopamine were investigated in a model of mechanical hyperalgesia. 2. Hyperalgesic responses to intraplantar (i.pl.) injections of carrageenin (100 micrograms), bradykinin (500 ng), TNF alpha (2.5 pg), IL-1 beta (0.5 pg), and IL-6 (1.0 ng), but not responses to IL-8 (0.1 ng), PGE2 (100 ng) and dopamine (10 micrograms), were inhibited by pretreatment with dexamethasone (0.5 mg kg-1, subcutaneously, s.c., or 0.04-5.0 micrograms/paw). 3. Inhibition of hyperalgesic responses to injections (i.pl.) of bradykinin (500 ng) and IL-1 beta (0.5 pg) by dexamethasone (0.5 mg kg-1, s.c.) was reversed by LCPS1 (0.5 ml kg-1, injected s.c., 24 h and 1 h before hyperalgesic substances) and hyperalgesic responses to injections (i.pl.) of bradykinin (500 ng), TNF alpha (2.5 pg) and IL-1 beta (0.5 pg), but not responses to PGE2 (100 ng), were inhibited by pretreatment with lipocortin 1(2-26) (100 micrograms/paw). Also, lipocortin-1(2-26) (30 and 100 micrograms ml 1 and dexamethasone (10 micrograms ml-1) inhibited TNF alpha release by cells of the J774 (murine macrophage-like) cell-line stimulated with LPS (3 micrograms ml 1), and LCPS1 partially reversed the inhibition by dexamethasone. These data are consistent with an important role for endogenous lipocortin-1(2-26) in mediating the anti-hyperalgesic effect of dexamethasone, with inhibiton of TNF alpha production by lipocortin-1(2-26) contributing, in part, to this role. 4. Although arachidonic acid by itself was not hyperalgesic, the hyperalgesic response to IL 1 beta (0.25 pg, i.pl.) was potentiated by arachidonic acid (50 micrograms) and the potentiated response was inhibited by dexamethasone (50 micrograms, i.pl.) and lipocortin-1(2-26) (100 micrograms, i.pl.). Also, lipocortin-1(2-26) (30 and 100 micrograms ml-1) inhibited/abolished PGE2 release by J774 cells stimulated with LPS (3 micrograms ml-1). These data suggest that, in inflammatory hyperalgesia, inhibition of the induction of cyclo-oxygenase 2 (COX-2), rather than phospholipase A2, by dexamethasone and lipocortin-1(2-26) accounts for the anti-hyperalgesic effects of these agents. 5. The above data support the notion that induction of lipocortin by dexamethasone plays a major role in the inhibition by dexamethasone of inflammatory hyperalgesia evoked by carrageenin, bradykinin and the cytokines TNF alpha, IL-1 beta and IL-6, and provides additional evidence that the biological activity of lipocortin resides within the peptide lipocortin-1(2-26). Further, the data suggest that inhibition of lipocortin-1(2-26) of eicosanoid production by COX-2 also contributes to the anti hyperalgesic effect of lipocortin-1. PMID- 9222545 TI - In vivo evidence for free radical involvement in the degeneration of rat brain 5 HT following administration of MDMA ('ecstasy') and p-chloroamphetamine but not the degeneration following fenfluramine. AB - 1. Administration of 3,4-methylenedioxymethamphetamine (MDMA or 'ecstasy') to several species results in a long lasting neurotoxic degeneration of 5 hydroxytryptaminergic neurones in several regions of the brain. We have now investigated whether this degeneration is likely to be the result of free radical induced damage. 2. Free radical formation can be assessed by measuring the formation of 2,3- and 2,5-dihydroxybenzoic acid (2,3-DHBA and 2,5-DHBA) from salicylic acid. An existing method involving implantation of a probe into the hippocampus and in vivo microdialysis was modified and validated. 3. Administration of MDMA (15 mg kg-1, i.p.) to Dark Agouti (DA) rats increased the formation of 2,3-DHBA (but not 2,5-DHBA) for at least 6 h. Seven days after this dose of MDMA, the concentration of 5-hydroxytryptamine (5-HT) and 5 hydroxyindoleacetic acid (5-HIAA) was reduced by over 50% in hippocampus, cortex and striatum, reflecting neurotoxic damage. There was no change in the concentration of dopamine or 3,4-dihydroxyphenylacetic acid (DOPAC) in the striatum. 4. p-Chloroamphetamine (PCA), another compound which produces a neurotoxic loss of cerebral 5-HT content, when given at a dose of 5 mg kg-1 also significantly increased the formation of 2.3-DHBA (but not 2,5-DHBA) in the dialysate for over 4.5 h. post-injection starting 2 h after treatment. 5. In contrast, fenfluramine administration (15 mg kg-1, i.p.) failed to increase the 2,3-DHBA or 2,5-DHBA concentration in the dialysate. A single fenfluramine injection nevertheless also markedly decreased the concentration of 5-HT and 5 HIAA in the hippocampus, cortex and striatum seven days later. 6. When rats pretreated with fenfluramine (15 mg kg-1, i.p.) seven days earlier were given MDMA (15 mg kg-1, i.p.) no increase in 2,3-DHBA was seen in the dialysate from the hippocampal probe. This indicates that the increase in free radical formation following MDMA is occurring in 5-HT neurones which have been damaged by the prior fenfluramine injection. 7. Administration of the free radical scavenging agent alpha-phenyl-N-tert-butyl nitrone (PBN; 120 mg kg-1, i.p.) 10 min before and 120 min after an MDMA (15 mg kg-1, i.p.) injection prevented the acute rise in the 2,3-DHBA concentration in the dialysate and attenuated by 30% the long term damage to hippocampal 5-HT neurones (as indicated by a smaller MDMA-induced decrease in both the concentration of 5-HT and 5-HIAA and also the binding of [3H]-paroxetine). 8. These data indicate that a major mechanism by which MDMA and PCA induce damage to 5-hydroxytryptaminergic neurones in rat brain is by increasing the formation of free radicals. These probably result from the degradation of catechol and quinone metabolites of these substituted amphetamines. In contrast, fenfluramine induces damage by another mechanism not involving free radicals; a proposal supported by some of our earlier indirect studies. 9. We suggest that these different modes of action render untenable the recent suggestion that MDMA will not be neurotoxic in humans because fenfluramine appears safe at clinical doses. PMID- 9222546 TI - Inhibition of monoamine oxidase A and B activities by imidazol(ine)/guanidine drugs, nature of the interaction and distinction from I2-imidazoline receptors in rat liver. AB - 1. I2-Imidazoline sites ([3H]-idazoxan binding) have been identified on monoamine oxidase (MAO) and proposed to modulate the activity of the enzyme through an allosteric inhibitory mechanism (Tesson et al., 1995). The main aim of this study was to assess the inhibitory effects and nature of the inhibition of imidazol(ine)/guanidine drugs on rat liver MAO-A and MAO-B isoforms and to compare their inhibitory potencies with their affinities for the sites labelled by [3H]-clonidine in the same tissue. 2. Competition for [3H]-clonidine binding in rat liver mitochondrial fractions by imidazol(ine)/guanidine compounds revealed that the pharmacological profile of the interaction (2-styryl-2 imidazoline, LSL 61112 > idazoxan > 2-benzofuranyl-2-imidazoline, 2-BFI = cirazoline > guanabenz > oxymetazoline > > clonidine) was typical of that for I2 sites. 3. Clonidine inhibited rat liver MAO-A and MAO-B activities with very low potency (IC50S: 700 microM and 6 mM, respectively) and displayed the typical pattern of competitive enzyme inhibition (lineweaver-Burk plots: increased K(m) and unchanged Vmax values). Other imidazol(ine)/guanidine drugs also were weak MAO inhibitors with the exception of guanabenz, 2-BFI and cirazoline on MAO-A (IC50S: 4-11 microM) and 2-benzofuranyl-2-imidazol (LSL 60101) on MAO-B (IC50: 16 microM). Idazoxan was a full inhibitor although with rather low potency, on both MAO-A and MAO-B isoenzymes (IC50S: 280 microM and 624 microM, respectively). Kinetic analyses of MAO-A inhibition by these drugs revealed that the interactions were competitive. For the same drugs acting on MAO-B the interactions were of the mixed type inhibition (increased K(m) and decreased Vmax values), although the greater inhibitory effects on the apparent value of Vmax/K(m) than on the Vmax value indicated that the competitive element of the MAO-B inhibition predominated. 4. Competition for [3H]-Ro 41-1049 binding to MAO A or [3H]-Ro 19-6327 binding to MAO-B in rat liver mitochondrial fractions by imidazol(ine)/guanidine compounds revealed that the drug inhibition constants (Ki values) were similar to the IC50 values displayed for the inhibition of MAO-A or MAO-B activities In fact, very good correlations were obtained when the affinities of drugs at MAO-A or MAO-B catalytic sites were correlated with their potencies in inhibiting MAO-A (r = 0.92) or MAO-B (r = 0.99) activity. This further suggested a direct drug interaction with the catalytic sites of MAO-A and MAO-B isoforms. 5. No significant correlations were found when the potencies of imidazol(ine)/guanidine drugs at the high affinity site (pKiH, nanomolar range) or the low-affinity site (pKiL, micromolar range) of I2-imidazoline receptors labelled with [3H]-clonidine were correlated with the pIC50 values of the same drugs for inhibition of MAO-A or MAO-B activity. These discrepancies indicated that I2-imidazoline receptors are not directly related to the site of action of these drugs on MAO activity in rat liver mitochondrial fractions. 6. Although these studies cannot exclude the presence of additional binding sites on MAO that do not affect the activity of the enzyme, they would suggest that I2-imidazoline receptors represent molecular species that are distinct from MAO. PMID- 9222547 TI - Characterization of [35S]-ATP alpha S and [3H]-alpha, beta-MeATP binding sites in rat brain cortical synaptosomes: regulation of ligand binding by divalent cations. AB - 1. We made a comparative analysis of the binding characteristics of the radioligands [35S]-ATP alpha S and [3H]-alpha, beta-MeATP in order to test whether these ligands can be used to analyse P2-purinoceptors in synaptosomal membranes from rat brain cortex. 2. Synaptosomes possess sites with high affinity for [35S]-ATP alpha S (Kd = 22.2 +/- 9.1 nM, Bmax = 14.8 pmol mg-1 protein). The rank order of the competition potency of the different compounds (ATP alpha S, ATP, ATP gamma S > ADP beta S, 2-MeSATP > deoxyATP, ADP > > UTP, alpha, beta MeATP, AMP, Reactive Blue-2, suramin, isoPPADS) is consistent with pharmacological properties of P2Y-purinoceptors. 3. Under identical conditions [35S]-ATP alpha S and [3H]-alpha, beta-MeATP bind to different binding sites at synaptosomal membranes from rat brain cortex. The affinity of the [3H]-alpha, beta-MeATP binding sites (Kd = 13.7 +/- 1.8 nM, Bmax = 6.34 +/- 0.28 pmol mg-1 protein) was 38 fold higher than the potency of alpha, beta-MeATP to displace [35S]-ATP alpha S binding (Ki = 0.52 microM). ATP and ADP beta S competed at both binding sites with different affinities, 60 fold and 175 fold, respectively. The other agonists tested (2-MeSATP, UTP, GTP) did not affect specific [35H]-alpha, beta-MeATP binding at concentrations up to 100 microM. The antagonists (suramin, isoPPADS, Evan's Blue) showed completely different affinities for both binding sites. 4. Binding of [35S]-ATP alpha S on synaptosomes was regulated by GTP, which is indicative for G-protein coupled receptors. The Kd value for the high affinity binding site was reduced in the presence of GTP about 5 fold (from 1.8 nM to 8.6 nM). In the presence of Mg2+ the affinity was increased (Kd 1.8 nM versus 22 nM in the absence of Mg2+). 5. The binding of both radioligands was regulated in an opposite manner by physiological concentrations of Ca2+ and Mg2+. Binding of [3H]-alpha, beta-MeATP to synaptosomal membranes was increased 3 fold by raising the Ca2+ concentration from 10 microM to 1 mM, whereas the addition of Mg2+ in the same concentration range resulted in an 80% reduction of the binding. In contrast, [35S]-ATP alpha S binding was not influenced at the same range of Ca2+ or Mg2+ concentrations (10 microM to 1 mM). The addition of Mg2+ (5 mM) increased the affinity of [35S]-ATP alpha S for the high affinity site 10 fold. 6. Diadenosine polyphosphates had a bimodal effect on [35S]-ATP alpha S binding to synaptosomal membranes. AP5A and Ap6A enhanced binding of [35S]-ATP alpha S 1.6 fold in a concentration range between 0.1 and 50 microM. Ap3A was a weak inhibitor with a Ki value of 7.2 microM. Ap4A, AP5A and Ap6A inhibited with Ki values > 100 microM. These data support the concept that diadenosine polyphosphates do not directly interact with ATP alpha S binding sites. 7. In conclusion, on the basis of present knowledge of the interaction of P2 purinoceptor active compounds with P2x- and/or P2Y-purinoceptors, our data strongly suggest that [35S]-ATP alpha S is a useful tool to study P2Y purinoceptors. Thus, the [35S]-ATP alpha S binding site might to a large extent represent P2Y-purinoceptors in synaptosomes from rat brain cortex. The nucleotide binding is regulated by G proteins, indicated by the effects of GTP/Mg2+ on binding. PMID- 9222548 TI - Neurochemical actions of the desglycinyl metabolite of remacemide hydrochloride (ARL 12495AA) in mouse brain. AB - 1. Remacemide hydrochloride, a recently developed antiepileptic drug, is believed to exert its effects, at least in part, via its desglycinyl metabolite, ARL 12495AA. 2. We have investigated the effects of ARL 12495AA on several neurochemical parameters in mouse brain. Adult male ICR mice were randomized into two groups and administered ARL 12495AA (0-75 mg kg-1) intraperitoneally, either as a single dose or once daily for 5 days. 3. Six hours after the final dose, animals were killed and their brains removed. Brain tissues were analysed for concentrations of gamma-aminobutyric acid (GABA), glutamine and glutamate and for the activities of GABA-transaminase (GABA-T) and glutamic acid decarboxylase (GAD). 4. Single dose ARL 12495AA was without effect on any of the parameters investigated. 5. Repeated ARL 12495AA treatment did not alter brain concentrations of GABA and glutamine, but at a high dose there was a trend toward reduced brain glutamate concentrations (P = 0.10). 6. Repeated administration of ARL 12495AA at a high dose significantly increased GABA-T activity (P < 0.05) and decreased that of GAD (P < 0.05). 7. These findings may have relevance to the clinical use of remacemide hydrochloride in human epilepsy. PMID- 9222549 TI - Carbon monoxide-induced vasorelaxation and the underlying mechanisms. AB - 1. Carbon monoxide (CO) induced a concentration-dependent relaxation of isolated rat tail artery tissues which were precontracted with phenylephrine or U-46619. This vasorelaxing effect of CO was independent of the presence of the intact endothelium. 2. The CO-induced vasorelaxation was partially inhibited by the blockade of either the cyclicGMP pathway or big-conductance calcium-activated K (KCa) channels. When both the cyclicGMP pathway and KCa channels were blocked, the CO-induced vasorelaxation was completely abolished. 3. Incubation of vascular tissues with hemin, in order to enhance the endogenous production of CO, suppressed the phenylephrine-induced vasocontraction in a time- and concentration dependent manner. The hemin-induced suppression of the vascular contractile response to phenylephrine was abolished after the vascular tissues were co incubated with either oxyhaemoglobin or zinc protoporphyrin-IX, suggesting an induced endogenous generation of CO from vascular tissues. 4. The effect of hemin incubation on vascular contractility did not involve the endogenous generation of nitric oxide. 5. Our results suggest that CO may activate both a cyclicGMP signalling pathway and KCa channels in the same vascular tissues, and that the endogenously generated CO may significantly affect the vascular contractile responses. PMID- 9222550 TI - Effect of chronic ETA-selective endothelin receptor antagonism on blood pressure in experimental and genetic hypertension in rats. AB - 1. Chronic treatment with a combined ETA/ETB endothelin receptor antagonist has been shown to reduce blood pressure in experimental rat models of hypertension in which endothelin-1 gene overexpression occurs in the walls of blood vessels, particularly small, resistance-sized arteries, but not in those genetic or experimental models of hypertension in which there is no overexpression of vascular endothelin-1. Failure of some experimental models of hypertension to respond to treatment with the combined ETA/ETB endothelin antagonist may be due in part to blockade of vasorelaxant endothelial ETB receptors which could in theory reduce the efficacy of endothelin antagonism. 2. In this study the orally active ETA-selective endothelin antagonists A-127722.5 and LU 135252 were used in chronic experiments on deoxycorticosterone acetate (DOCA)-salt hypertensive rats (which overexpress vascular endothelin-1 and respond with blood pressure lowering to combined ETA/ETB endothelin receptor antagonism), on spontaneously hypertensive rats (SHR) (which do not overexpress vascular endothelin-1 and do not respond with blood pressure lowering to the combined ETA/ETB receptor antagonist), and in 1-kidney 1 clip Goldblatt (1-K IC) hypertensive rats (which present mild overexpression of vascular endothelin-1 but do not respond with blood pressure lowering to the combined ETA/ETB receptor antagonist). Additionally, it has been suggested that interruption of the renin-angiotensin system may sensitize responses to endothelin antagonism. Accordingly, SHR were treated with an angiotensin converting enzyme inhibitor, cilazapril, in addition to the ETA receptor antagonist. 3. Blood pressure of DOCA-salt hypertensive rats was lowered by a mean of 24 and of 27 mmHg (P < 0.01) by A-127722.5 after 4 weeks of treatment, when given orally at two different doses (10 and 30 mg kg-1 day-1), and by 18 mmHg by LU 135252 50 mg kg-1 day-1. 4. SHR treated with A-127722.5 for 8 weeks starting at 12 weeks of age exhibited the same progressive rise in blood pressure as untreated SHR. Addition of cilazapril resulted in similar reduction of blood pressure in A-127722.5-treated and untreated SHR. 5. Treatment of 1-K IC hypertensive rats with the dose of LU 135252 which lowered blood pressure in DOCA salt hypertensive rats did not cause any reduction in blood pressure relative to untreated rats. 6. These results demonstrate that treatment with either dose of the selective ETA receptor antagonists A-127722.5 or LU 135252 caused reductions in blood pressure similar to those obtained for a combined ETA/ETB endothelin antagonist. Blood pressure was lowered only in hypertensive rats known to overexpress vascular endothelin-1 (DOCA-salt hypertensive rats) but not in those which do not (SHR) or only have mild vascular overexpression of endothelin-1 gene (1-K 1C hypertensive rats). Reduction in activity of the renin-angiotensin system in SHR did not sensitize blood pressure to potential hypotensive effects of an ETA-selective receptor antagonist. PMID- 9222551 TI - Effect of a selective 5-HT reuptake inhibitor in combination with 5-HT1A and 5 HT1B receptor antagonists on extracellular 5-HT in rat frontal cortex in vivo. AB - 1. Selective 5-hydroxytryptamine (5-HT; serotonin) reuptake inhibitors (SSRIs) cause a greater increase in extracellular 5-HT in the forebrain when the somatodendritic 5-HT1A autoreceptor is blocked. Here, we investigated whether blockade of the terminal 5-HT1B autoreceptor influences a selective 5-HT reuptake inhibitor in the same way, and whether there is an additional effect of blocking both the 5-HT1A and 5-HT1B autoreceptors. 2. Extracellular 5-HT was measured in frontal cortex of the anaesthetized rat by use of brain microdialysis. In vivo extracellular recordings of 5-HT neuronal activity in the dorsal raphe nucleus (DRN) were also carried out. 3. The selective 5-HT reuptake inhibitor, paroxetine (0.8 mg kg-1, i.v.), increased extracellular 5-HT about 2 fold in rats pretreated with the 5-HT1A receptor antagonist, WAY100635. When administered alone neither paroxetine (0.8 mg kg-1, i.v.) nor WAY100635 (0.1 mg kg-1, i.v.) altered extracellular 5-HT levels. 4. Paroxetine (0.8 mg kg-1, i.v.) did not increase 5 HT in rats pretreated with the 5-HT1B/D receptor antagonist, GR127935 (1 mg kg-1, i.v.). GR127935 (1 and 5 mg kg-1, i.v.) had no effect on extracellular 5-HT when administered alone. 5. Interestingly, paroxetine (0.8 mg kg-1, i.v.) caused the greatest increase in 5-HT (up to 5 fold) when GR127935 (1 or 5 mg kg-1, i.v.) was administered in combination with WAY100635 (0.1 mg kg-1, i.v.). Administration of GR127935 (5 mg kg-1, i.v.) plus WAY100635 (0.1 mg kg-1, i.v.) without paroxetine, had no effect on extracellular 5-HT in the frontal cortex. 6. Despite the lack of effect of GR127935 on 5-HT under basal conditions, when 5-HT output was elevated about 3 fold (by adding 1 microM paroxetine to the perfusion medium), the drug caused a dose-related (1 and 5 mg kg-1, i.v.) increase in 5-HT. 7. By itself, GR127935 slightly but significantly decreased 5-HT cell firing in the DRN at higher doses (2.0-5.0 mg kg-1, i.v.), but did not prevent the inhibition of 5-HT cell firing induced by paroxetine. 8. In summary, our results suggest that selective 5-HT reuptake inhibitors may cause a large increase in 5-HT in the frontal cortex when 5-HT autoreceptors on both the somatodendrites (5-HT1A) and nerve terminals (5-HT1B) are blocked. This increase is greater than when either set of autoreceptors are blocked separately. The failure of a 5-HT1B receptor antagonist alone to enhance the effect of the selective 5-HT reuptake inhibitor in our experiments may be related to a lack of tone on the terminal 5-HT1B autoreceptor due to a continued inhibition of 5-HT cell firing. These results are discussed in relation to the use of 5-HT autoreceptor antagonists to augment the antidepressant effect of selective 5-HT reuptake inhibitors. PMID- 9222552 TI - Interaction of a new bis-indol derivative, KAR-2 with tubulin and its antimitotic activity. AB - 1. KAR-2 (3"-(beta-chloroethyl)-2",4"-dioxo-3,5"-spiro-oxazolidino- 4-deacetoxy vinblastine), is a bis-indol derivative; catharantine is coupled with the vindoline moiety which contains a substituted oxazolidino group. Our binding studies showed that KAR-2 exhibited high affinity for bovine purified brain tubulin (Kd-3 microM) and it inhibited microtubule assembly at a concentration of 10 nM. 2. Anti-microtubular activity of KAR-2 was highly dependent on the ultrastructure of microtubules: while the single tubules were sensitive, the tubules cross-linked by phosphofructokinase (ATP: D-fructose-6-phosphate-1 phosphotransferase, EC 2.7.1.11) exhibited significant resistance against KAR-2. 3. The cytoplasmic microtubules of Chinese hamster ovary mammalian and Sf9 insect cells were damaged by 1 microgram ml-1 KAR-2, as observed by indirect immunofluorescence and transmission electron microscopy. Scanning electron microscopy revealed intensive surface blebbing on both types of cells in the presence of KAR-2. 4. KAR-2 was effective in the mouse leukaemia P338 test in vivo without significant toxicity. Studies on a primary cerebro-cortical culture of rat brain and differentiated PC12 cells indicated that the toxicity of KAR-2 was significantly lower than that of vinblastine. The additional property of KAR 2 that distinguishes it from bis-indol derivatives is the lack of anti-calmodulin activity. PMID- 9222553 TI - The interaction of a new anti-tumour drug, KAR-2 with calmodulin. AB - 1. KAR-2 (3"-(beta-chloroethyl)-2",4"-dioxo-3,5" -spiro-oxazolidino-4-deacetoxy vinblastine) is a semisynthetic bis-indol derivative, with high anti-microtubular and anti-tumour activities but with low toxicity. KAR-2, in contrast to other biologically active bis-indols (e.g. vinblastine) did not show anti-calmodulin activity in vitro (enzyme kinetic, fluorescence anisotropy and immunological tests). 2. Direct binding studies (fluorescence resonance energy transfer, circular dichroism) provided evidence for the binding of KAR-2 to calmodulin. The binding affinity of KAR-2 to calmodulin (dissociation constant was about 5 microM) in the presence of Ca2+ was comparable to that of vinblastine. 3. KAR-2 was able to interact with apo-calmodulin as well; in the absence of Ca2+ the binding was of cooperative nature. 4. The effect of drugs on Ca2+ homeostasis in human neutrophil cells was investigated by means of a specific fluorescent probe. Trifluoperazine extensively inhibited the elevation of intracellular Ca2+ level, vinblastine did not appreciably affect it, KAR-2 stimulated the Ca2+ influx and after a transient enhancement the Ca2+ concentration reached a new steady-state level. 5. Comparison of the data obtained with KAR-2 and bis-indols used in chemotherapy suggests that the lack of anti-calmodulin potency resides on the spiro-oxazolidino portion of KAR-2. This character of KAR-2 manifested itself in various systems and might result in its low in vivo toxicity, established in an anti-tumour test. PMID- 9222554 TI - Identification and pharmacological characterization of somatostatin receptors in rat lung. AB - 1. [125I]-[LTT]SRIF-28 and [125I]-SMS 201-995 were used to identify and characterize somatostatin (SRIF) receptors localized in rat lung tissue. In vitro autoradiography of rat lung tissue sections showed the existence of specific, high affinity binding sites for [125I]-[LTT]SRIF-28 without any significant specific binding of the sst2/sst5-receptor selective ligand [125I]-SMS 201-995. 2. In radioligand binding studies, specific binding of [125I]-[LTT]SRIF-28 to membranes of rat lung was linearly related to the concentration of membrane protein used with only a small portion of nonspecific binding. With [125I]-SMS 201-995 no specific binding could be observed up to a membrane concentration of 0.1 mg of protein/assay tube. 3. [125I]-[LTT]SRIF-28 bound rapidly to rat lung membranes with an apparent association rate constant (kapp) of 1.8 +/- 0.1 h-1 (n = 3). The equilibrium of specific binding was reached after an incubation period of approximately 90 min at room temperature and remained constant for the next 3 h. The association rate constant (k1) was calculated to be 3.7 x 10(10) M-1 h-1. The dissociation reaction followed first order kinetics with a dissociation rate constant (k-1) = 0.44 +/- 0.07 h-1 corresponding to a half-time of 95 +/- 15 min (n = 3). From these kinetic experiments an equilibrium dissociation constant (KD) for the binding of [125I]-[LTT]SRIF-28 was calculated to be 11.9 pM. 4. Saturation binding of [125I]-[LTT]SRIF-28 revealed an equilibrium dissociation constant (KD) of 50.1 pM (pKD = 10.3 +/- 0.1; n = 3) and a receptor density (Bmax) of 78 +/- 3 fmol mg-1 protein. A Hill coefficient not significantly different from 1 indicated saturable binding to a single class of high affinity binding sites. 5. Specific binding of [125I]-[LTT]SRIF-28 to rat lung membranes was inhibited by SRIF-14, SRIF-28 and different SRIF analogues. SRIF and different synthetic short chain SRIF analogues exhibited the following rank order of potency: SRIF-28 > SRIF-14 > CGP 23996 >> RC 160 > BIM 23014 > SMS 201- 995 > BIM 23056 > MK 678. 6. The binding affinities for SRIF and the various SRIF analogues determined using rat lung tissue were in close correlation to those obtained with Chinese hamster ovary (CHO) cells stably expressing sst, (r = 0.92) and sst4 (r = 0.95) receptors, respectively. 7. Reverse transcriptase--polymerase chain reaction (RT-PCR) showed the predominant expression of mRNA specific for sst4 receptors as well as some weak sst1 mRNA expression. 8. The findings suggest that sst4 receptor expression is the predominant form of the somatostatin receptors identified in rat lung tissue. In this study we demonstrated for the first time the existence of sst4 receptors in mammalian tissue. PMID- 9222555 TI - Impairment of endothelium-dependent but not of endothelium-independent dilatation in guinea-pig aorta rings incubated in the presence of elevated glucose. AB - 1. Purine compounds such as ATP and adenosine, respectively endothelium-dependent and- independent vasodilators, are largely involved in the control of vascular tone and vascular reactivity to contracting stimuli. We investigated the relaxing activity of ATP and adenosine in guinea-pig aorta rings exposed for 6 h to elevated glucose concentration (50 mM), in order to mimic hyperglycaemic conditions. Guinea-pigs were reserpine-treated (2 mg kg-1, i.p., 48 and 24 h before death). 2. Rings of aortae incubated in 50 mM glucose, contracted submaximally by 1 microM noradrenaline, lost endothelium-dependent relaxation in response to acetylcholine (10 nM to 10 microM). Aortae incubated with 50 mM mannose, as a hyperosmotic control, relaxed to acetylcholine normally. Rings of aortae incubated in 50 mM glucose, contracted submaximally by 3 mM 4 aminopyridine, lost endothelium-dependent relaxation in response to ATP (30 microM) whereas endothelium-independent relaxation in response to adenosine (0.3 mM) was well preserved. 4. The relaxation induced by A23187 or sodium nitroprusside (10 nM to 0.1 microM) did not differ between rings exposed to control (5.5 mM) or elevated glucose (50 mM) and contracted submaximally by 3 mM 4-aminopyridine. 5. When incubated with aortic tissue in the presence of elevated glucose, the cyclo-oxygenase inhibitors, indomethacin (10 microM) and mefenamic acid (30 microM), or the scavenger of superoxide anions, superoxide dismutase (150 u ml-1), prevented the impairment of ATP-mediated relaxation. 6. The present results indicate that endothelium-dependent, receptor-induced relaxation in response to acetylcholine and ATP is impaired in guinea-pig aorta rings exposed to elevated glucose. The endothelial dysfunction caused by glucose might be located at a step between receptor activation and intracellular calcium increase, and might be related to an increased metabolism of arachidonic acid coupled to an increased production, or to a reduced inactivation of superoxide anions. PMID- 9222556 TI - The mechanisms of the relaxation induced by vasoactive intestinal peptide in the porcine coronary artery. AB - 1. This study was designed to investigate the mechanism of the relaxation induced by vasoactive intestinal peptide (VIP) in medial strips of the porcine coronary artery, by determining the effect on the cytosolic Ca2+ concentration ([Ca2+]i), the [Ca2+]i-force relation and the involvement of G-protein. 2. Front-surface fluorometry of fura-2 revealed that U46619, a thromboxane A2 analogue, and the high K(+)-depolarization induced increases in both the [Ca2+]i and force of the medial strips. At a steady state of contraction, the extent of an increase in [Ca2+]i induced by 100 nM U46619 was similar to that induced by 30 mM K(+) depolarization. VIP concentration-dependently (1 nM-1 microM) induced transient decreases in both the [Ca2+]i and force of the medial strips precontracted with 100 nM U46619. The decreases in the [Ca2+]i and force induced by VIP during the contraction with U46619 were much greater than those with 30 mM K(+) depolarization. 3. The VIP-induced decreases in the [Ca2+]i and force were attenuated by K+ channel blockers such as tetrabutylammonium (TBA: non-selective K+ channel blocker), charybdotoxin (large conductance Ca(2+)-activated K+ channel blocker), and 4-aminopyridine (4-AP: voltage-dependent K+ channel blocker). However, neither glibenclamide (ATP-sensitive K+ channel blocker) nor apamin (small conductance Ca(2+)-activated K+ channel blocker) had any significant inhibitory effect. 4. In the 30 mM K(+)-depolarized strips, pretreatment with thapsigargin, a specific Ca(2+)-ATPase inhibitor of the Ca2+ store sites, completely abolished the VIP-induced decrease in [Ca2+]i, but partially attenuated the VIP-induced decrease in force. 5. VIP shifted the [Ca2+]i-force relation of the U46619-induced contractions to the right in a concentration dependent manner. In the alpha-toxin-permeabilized strips, VIP decreased the force development at a constant [Ca2+]i level (pCa = 6.5) in a GTP-dependent manner, which was antagonized by guanosine-5'-O-(beta-thiodiphosphate) (GDP beta S). 6. We thus conclude that VIP relaxes the coronary artery via three mechanisms: (1) a decrease in [Ca2+]i by inhibiting the Ca2+ influx presumably through the membrane hyperpolarization mediated by the activation of the large conductance Ca(2+)-activated (charybdotoxin-sensitive) K+ channels and voltage dependent (4-AP-sensitive) K+ channels; (2) a decrease in [Ca2+]i by sequestrating cytosolic Ca2+ into thapsigargin-sensitive Ca2+ store sites; and (3) a decrease in the Ca(2+)-sensitivity of the contractile apparatus through the activation of G-protein. PMID- 9222557 TI - Cholinesterase activity in human pulmonary arteries and veins. AB - 1. Human isolated pulmonary vessels were treated with cholinesterase (ChE) inhibitors to determine the role of these enzymes in regulating vascular muscle tone. In addition, kinetic parameters were determined for acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in human pulmonary vessel homogenates. 2. Carbachol (CCh) and acetylcholine (ACh) were equipotent contractile agonists in human pulmonary arteries (pD2 values, 5.28 +/- 0.05 and 5.65 +/- 0.16; Emax, 0.91 +/- 0.26 and 0.98 +/- 0.30 g wt. for CCh and ACh, respectively; n = 7). In venous preparations, ACh was ineffective and CCh induced small contractions (Emax, 0.08 +/- 0.04 g wt; n = 13). 3. In human pulmonary arteries following pretreatment with tetraisopropylpyrophosphoramide (iso-OMPA, 100 microM), an increased sensitivity to the contractile agonist ACh was observed (pD2 values, 5.80 +/- 0.13 and 6.37 +/- 0.19 for control and treated preparations, respectively; n = 5). This pretreatment had no effect on the CCh concentration response curve. In contrast, human pulmonary veins pretreated with iso-OMPA failed to elicit a contractile response to ACh. 4. Neither Iso-OMPA nor neostigmine elicited concentration-dependent contractions in human isolated pulmonary arteries or veins. These results suggest the absence of sufficient spontaneous release of ACh to modulate human pulmonary vessel basal tone. 5. CCh was less potent than ACh in relaxing precontracted human isolated pulmonary arteries (pD2 value, CCh: 6.55 +/ 0.15 and ACh: 7.16 +/- 0.13, n = 4) and veins (pD2 value, CCh: 4.95 +/- 0.13; n = 5 and ACh: 5.56 +/- 0.17; n = 6). Pretreatment of vessels with either iso-OMPA or neostigmine did not modify ACh relaxant responses in either type of preparation. 6. In human pulmonary veins, the ChE activity was two fold greater than in arteries (n = 6). Vmax for AChE was 1.73 +/- 0.24 and 3.36 +/- 0.26 miu mg-1 protein in arteries and veins, respectively, whereas Vss for BChE was 1.83 +/- 0.22 and 4.71 +/- 0.17 miu mg-1 protein, in these respectively. 7. In human pulmonary arteries, BChE activity may play a role in the smooth muscle contraction but not on the smooth muscle endothelium-dependent relaxation induced by ACh. A role for ChE activity in the control of venous tone is presently difficult to observe, even though this tissue contains a greater amount of enzyme than the artery. PMID- 9222558 TI - Involvement of calyculin A inhibitable protein phosphatases in the cyclic AMP signal transduction pathway of mouse corticotroph tumour (AtT20) cells. AB - 1. The role of non-calcineurin protein phosphatases in the cyclic AMP signal transduction pathway was examined in mouse pituitary corticotroph tumour (AtT20) cells. 2. Blockers of protein phosphatases, calyculin A and okadaic acid, were applied in AtT20 cells depleted of rapidly mobilizable pools of intracellular calcium and activated by various cyclic AMP generating agonists. Inhibitors of cyclic nucleotide phosphodiesterases were present throughout. The accumulation of cyclic AMP was monitored by radioimmunoassay, phosphodiesterase activity in cell homogenates was measured by radiometric assay. 3. Neither calyculin A nor okadaic acid altered basal cyclic AMP levels but cyclic AMP formation induced by 41 amino acid residue corticotrophin releasing-factor (CRF) was strongly inhibited (up to 80%), 1-Norokadaone was inactive. Similar data were also obtained when isoprenaline or pituitary adenylate cyclase activating peptide1-38 were used as agonists. 4. Pertussis toxin did not modify the inhibition of CRF-induced cyclic AMP production by calyculin A. 5. Pretreatment with calyculin A completely prevented the stimulation of cyclic AMP formation by cholera toxin even in the presence of 0.5 mM isobutylmethylxanthine (IBMX) and 0.1 mM rolipram. Cholera toxin mediated ADP-ribosylation of the 45 K and 52 K molecular weight Gs alpha isoforms in membranes from calyculin A-pretreated cells was enhanced to 150-200% when compared with controls. 6. Cholera toxin-induced cyclic AMP was reduced by calyculin A within 10 min when calyculin A was applied after a 90 min pretreatment with cholera toxin. Under these conditions the effect of calyculin A could be blocked by the combination of 0.5 mM IBMX and 0.1 mM rolipram, but not by 0.5 mM IBMX alone. 7. Phosphodiesterase activity in AtT20 cell homogenates showed a significant, 2.7 fold increase after treatment with calyculin A. In control cells phosphodiesterase activity was blocked by 80% in the presence of IBMX (0.5 mM), or IBMX plus rolipram (0.1 mM). In calyculin A-treated cells phosphodiesterase activity was also strongly inhibited by IBMX, but because of the stimulating effect of calyculin A, the activity remaining was still 55% of that found in control homogenates. This activity was reduced to 5% of control by using IBMX and rolipram in combination. Assay of phosphodiesterase in Ca2+ free conditions showed that calyculin A markedly increases the activity of rolipram sensitive (type 4) phosphodiesterase. 8. Taken together, blockers of protein phosphatases (PPases) impaired signal transduction through Gs-mediated pathways and activated cyclic AMP degrading phosphodiesterase(s), indicating that PPases 1 and/or 2A are essential for agonist-mediated regulation of cyclic AMP levels in AtT20 cells, and are thus important in maintaining the secretory phenotype of the cells. PMID- 9222559 TI - The selectivity and structural determinants of peptide antagonists at the CGRP receptor of rat, L6 myocytes. AB - 1. Potency orders were determined for a series of agonists and antagonists on the calcitonin gene-related peptide (CGRP) receptor of rat L6 myocytes. The agents tested were all shown to have been active against CGRP, amylin or adrenomedullin receptors. 2. AC187 had a pIC50 of 6.8 +/- 0.10, making it 14 fold less potent as an antagonist than CGRP8-37 (pIC50, 7.95 +/- 0.14). Amyline8-37 was equipotent to AC187 (pIC50, 6.6 +/- 0.16) and CGRP19-32 was 3 fold less potent than either (pIC50, 6.1 +/- 0.24). 3. [Ala11]-CGRP8-37 was 6 fold less potent than CGRP8-37, (pIC50, 7.13 +/- 0.14), whereas [Ala18]-CGRP8-37 was approximately equipotent to CGRP8-37 (pIC50, 7.52 +/- 0.15). However, [Ala11,Ala18]-CGRP8-37 was over 300 fold less potent than CGRP8-37 (pIC50, 5.30 +/- 0.04). 4. [Tyr0]-CGRP28-37, amylin19-37 and adrenomedullin22-52 were inactive as antagonists at concentrations of up to 1 microM. 5. Biotinyl-human alpha-CGRP was 150 fold less potent than human alpha-CGRP itself (EC50 values of 48 +/- 17 nM and 0.31 +/- 0.13 nM, respectively). At 1 microM, [Cys(acetomethoxy)2,7]-CGRP was inactive as an agonist. 6. These results confirm a role for Arg11 in maintaining the high affinity binding of CGRP8-37. Arg18 is of less direct significance for high affinity binding, but it may be important in maintaining the amphipathic nature of CGRP and its analogues. PMID- 9222560 TI - Induction of bradykinin B1 receptors in rat colonic epithelium. AB - 1. Des-Arg9 bradykinin (DAB), a classical B1-kinin receptor agonist was without effect when applied to the basolateral surface of rat isolated colon epithelium. Three hours after tissues were isolated DAB caused, after a delay of up to 2 min, a maintained increase of short circuit current (SCC). 2. The SCC increase in colonic epithelia, mounted in vitro for three hours, caused by DAB was due to electrogenic chloride secretion as the current increase was reversed by frusemide and did not occur in the absence of cystic fibrosis transmembrane conductance regulator (CFTR) chloride channels. The EC50 for DAB was approximately 50 nM. 3. An inhibitor of transcription (actinomycin D) and of translation (cycloheximide) prevented the appearance of DAB sensitivity without affecting the responses to another secretagogue (forskolin). 4. The classical B1-kinin receptor antagonist, Leu8-des-Arg9 bradykinin, was shown to be an agonist in rat colon epithelium. Other B1-kinin receptor antagonists (des-Arg10-Hoe 140 and R-715) inhibited the responses to DAB in 'aged' colonic epithelia, and the inhibition was easily surmounted by increasing the concentration of DAB. 5. Response to DAB did not appear to involve to any significant extent, the formation of prostaglandins, leukotrienes, histamine or nitric oxide. Furthermore, no neuronal involvement was apparent in the stripped colonic preparations. The responses to DAB were not significantly different in epithelia taken from different parts of the distal colon. 6. The differences between the responses of the colonic epithelium to B1- and B2-kinin receptor agonists are discussed. PMID- 9222561 TI - Pharmacological characterization of the vanilloid receptor in the rat dorsal spinal cord. AB - 1. In the present study a novel 96-well plate assay system was used to characterize pharmacologically the vanilloid receptor in the dorsal spinal cord of the rat. When activated, this receptor stimulates release of calcitonin gene related peptide (CGRP) from the central terminals of the afferent nerves. 2. Capsaicin, resiniferatoxin (RTX) and olvanil each evoked a concentration dependent increase in CGRP release with pEC50 values of 6.55 +/- 0.07, 7.90 +/- 0.24 and 6.19 +/- 0.15 respectively. RTX and olvanil were partial agonists with respect to capsaicin. All concentration-effect curves were bell-shaped. 3. The vanilloid receptor antagonist, capsazepine (10 microM) had no effect on basal peptide release but inhibited the CGRP release evoked by all 3 agonists to a similar extent. These results suggest that the antagonistic effects of capsazepine were agonist-independent. 4. The capsaicin-sensitive cation channel blocker, ruthenium red (10 microM) had no effect on basal CGRP release, but antagonized the peptide release evoked by capsaicin, olvanil and RTX. 5. The pharmacology of the vanilloid receptor in the rat dorsal spinal cord is not identical to that previously found in other systems. The reason for these differences is unclear, but the possibility of multiple classes of receptor cannot at this stage be ruled out. PMID- 9222563 TI - The involvement of ATP-sensitive potassium channels in beta 2-adrenoceptor agonist-induced vasodilatation on rat diaphragmatic microcirculation. AB - 1. The effects of glibenclamide (GLB), a blocker of ATP-sensitive potassium (KATP) channels, on diaphragmatic microcirculation in male Sprague-Dawley rats were assessed under basal conditions and after beta 2-adrenoceptor-agonist stimulation. In addition, forskolin was used to bypass beta-adrenoceptors and GTP binding proteins (G-protein) to explore the possible mechanism of GLB effects. For comparison, the relationships between KATP channel activity and cyclic GMP mediated vasodilator responses to acetylcholine (ACh) and sodium nitroprusside (SNP) were also assessed. 2. Male Sprague-Dawley rats were anaesthetized with urethane and mechanically ventilated. The left hemi-diaphragm of each rat was prepared and microvascular blood flow (QLDF) was recorded with laser-Doppler flowmetry during continuous superfusion with bicarbonate-buffered, prewarmed Ringer solution. The drugs were topically applied to the surface of the hemi diaphragm. 3. Salbutamol (0.32-32 microM), terbutaline (0.32 microM-0.32 microM) and forskolin (0.32-10 microM) each elicited a concentration-dependent increase in QLDF. 4. Baseline microvascular blood flow was unaffected by a 30 min suffusion of 1 microM GLB (295 +/- 51 mV vs 325 +/- 62 mV. P = 0.738). 5. The vasodilator response elicited by salbutamol (0.32 microM, 1 microM and 3.2 microM and 10 microM), was significantly attenuated by a 30 min superfusion with 1 microM GLB; this salbutamol-induced vasodilatation was mediated via an interaction with beta-adrenoceptor receptors, as in other experiments it was greatly inhibited by 30-min superfusion with propranolol (10 microM). 6. Similarly, following 30-min superfusion with GLB (1 microM), the terbutaline (1 microM, 3.2 microM and 10 microM)-induced vasodilator response was almost abolished and the vasodilator responses induced by incremental concentrations of forskolin (0.32 microM, 1 microM and 3.2 microM) were also significantly attenuated. 7. Cromakalim (1.5 microM, 3 microM and 3.2 microM) produced an increase of QLDF in a dose-dependent manner, which was virtually abolished by GLB (1 microM). In contrast, the vasodilator responses induced by acetylcholine (32 microM, 0.1 mM, and 0.32 mM) or sodium nitroprusside (3.2 microM, 10 microM and 20 microM) were independent of GLB (1 microM). 8. In conclusion, KATP channels may be functional, but tonically inactive in the resting diaphragmatic microcirculation and the vasodilator effect of beta 2-adrenoceptor agonists may be partly mediated by KATP channels; the activation of KATP channels may involve the accumulation of cyclic AMP in vascular smooth muscle cells. PMID- 9222562 TI - Alpha 1-adrenoceptor vasoconstriction in the tail artery during ageing. AB - 1. We have studied the alpha 1-adrenoceptor-mediated responses in intact tail artery rings from 3-4 and 20-22 months old Sprague-Dawley rats, focusing on possible endothelial alterations. The influence of nitric oxide released by the endothelium, the number of alpha 1-adrenoceptors and the functional receptor reserve were evaluated to determine their contribution to the contractile response mediated by this receptor. The state of the endothelial layer was assessed by confocal microscopy. 2. Noradrenaline (1 nM-100 microM) induced concentration-dependent vasoconstriction. The maximum contractions to noradrenaline (P < 0.05) and to 75 mM KCl (P < 0.01) were higher in young than in old animals. 3. The density (Bmax) of alpha 1-adrenoceptors and the dissociation constant (KD) obtained in [3H]-prazosin binding experiments were unchanged by age. 4. The apparent affinity (pKA) and the percentage of functional receptors (qx 100) remaining after phenoxybenzamine (0.03 microM) were similar in both age groups. 5. After partial alpha 1-adrenoceptor inactivation with phenoxybenzamine, NG-nitro-L-arginine methylester (30 microM) significantly potentiated the E/[A] curve to noradrenaline in young rats. However, only responses to 0.1 to 1 microM noradrenaline were significantly potentiated in old animals. In addition, 94% of the vessels from young, but only 52% from old rats were relaxed by 80-100% of the noradrenaline (0.03 microM) contraction, with 1 microM acetylcholine. 6. No modifications in the area (micron2) or in the number of endothelial nuclei (per mm2) were observed between age groups. An elongation of the nuclei of endothelial cells was observed in the old animals. 7. These data suggest that the noradrenaline-induced contraction is decreased in old rats probably due to differences in either the contractile machinary or postreceptor mechanisms. These alterations may be accompanied by an impairment of the release or production of NO from endothelial cells. PMID- 9222566 TI - Family medicine residency programs. Do they encourage lifelong learning? PMID- 9222564 TI - Nitric oxide-dependent and -independent vascular hyporeactivity in mesenteric arteries of portal hypertensive rats. AB - 1. Increased production of nitric oxide (NO) has been suggested to underlie both the vascular hyporeactivity to vasoconstrictors and the splanchnic vasodilatation seen in portal hypertension. This study assessed the role of NO in the vasoconstrictor hyporeactivity of portal vein-ligated (PVL) rats in isolated and in situ perfused mesenteric arterial beds. 2. Isolated perfused mesenteric arteries of PVL rats were significantly less reactive to noradrenaline (NA), methoxamine (METH), arginine vasopressin (AVP) and endothelin-1 (ET-1) than those from sham-operated (Sham) rats. 3. Blockade of NO synthesis with NG-nitro-L arginine methyl ester (L-NAME, 100 microM) in isolated perfused mesenteric arteries from PVL rats restored the reactivity to bolus injections of AVP and ET 1, but had little effect on the hyporeactivity to NA or METH. Cyclo-oxygenase inhibition with indomethacin (5 microM) likewise did not restore reactivity to METH of isolated perfused mesenteric arteries of PVL rats. 4. The hyporeactivity to METH seen in isolated perfused mesenteric arteries from PVL rats was reduced by low concentrations of AVP (20 nM) or ET-1 (1 nM) which per se caused only a slight increase in perfusion pressure. When L-NAME (100 microM) was combined with AVP (20 nM) or ET-1 (1 nM), respectively, reactivity to METH of isolated perfused mesenteric arteries of PVL rats was restored to the level seen in Sham rats. These effects of AVP and ET-1 were not mimicked by precontracting the vessels with 5-hydroxytryptamine (5 microM). 5. The differential effects of L-NAME and AVP on the hyporesponsiveness to methoxamine and AVP were corroborated by experiments performed with the in situ perfused mesenteric vascular bed preparation. 6. These data indicate that both NO-dependent and NO-dependent mechanisms are involved in the vasoconstrictor hyporesponsiveness of mesenteric arteries from portal hypertensive rats. The hyporeactivity to AVP and ET-1 is mediated by NO whereas the reduced responsiveness to adrenoceptor agonists appears to be predominantly NO-independent AVP and ET-1, in addition, seem to inhibit the NO-independent mechanism of vascular hyporeactivity, since the hyporesponsiveness to METH was reduced in the presence of AVP or ET-1 and abolished by the combination of these peptides with L-NAME. PMID- 9222565 TI - Lack of involvement of glutamate-induced excitotoxicity in MPP+ toxicity in striatal dopaminergic terminals: possible involvement of ascorbate. AB - 1. The present study concerns the possible relationship between glutamate excitotoxicity and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine/1-methyl-4 phenylpyridinium (MPTP/MPP+) neurotoxicity on striatal dopaminergic terminals. 2. MPP+ neurotoxicity has been studied by means of two MPP+ perfusions separated by 24 h. After the second MPP+ 1 mM perfusion, dopamine extracellular output, measured by microdialysis, was considered to be an index of the dopaminergic neurone damage produced by the first MPP+ 1 mM perfusion. 3. High concentration (10 mM) of glutamate uptake inhibitor L-trans-pyrrolidine-2,4-dicarboxylic acid (PDC) stimulated basal release of dopamine and protected against the neurotoxic effect of MPP+. 4. PDC 10 mM perfusion produced an increase in the extracellular output of glutamate and aspartate, and a decrease in that of ascorbate. 5. The protective effect against MPP+ toxicity observed with PDC 10 mM was completely abolished when this glutamate uptake inhibitor was co-perfused with ascorbate 0.5 mM. 6. These results suggest that glutamate-induced neurotoxicity is not involved in MPP+ toxicity. The protective effect found with the glutamate uptake inhibitor could be due to a decrease in extracellular ascorbate levels. PMID- 9222567 TI - Resource allocation and ethics. Can clinicians find a middle ground? PMID- 9222568 TI - General practice training in Uganda. Part 1: Setting, personnel, and facilities. AB - Making lasting changes to health care in developing countries is a challenge to industrialized nations. This two-part article outlines how a general practice training project was established in Uganda to prepare postgraduate students to work in rural hospitals. Part 1 covers the setting, those involved, and the facilities available to launch such a project. PMID- 9222569 TI - General practice training in Uganda. Part 2: Training program and clinical practice. AB - Enabling a developing country to become self-sufficient in health care is a major feat. This two-part article outlines how a general practice training project was established in Uganda to prepare postgraduate students to work in rural hospitals. The first part looked at the setting, personnel, and facilities. Part 2 outlines the curricula and clinical practice. PMID- 9222570 TI - No room to assume in birth control. PMID- 9222571 TI - Benzodiazepine use by students: risky business. PMID- 9222572 TI - Concerns about formula advertising. PMID- 9222573 TI - Can pregnant patients safely take nitrofurantoin? PMID- 9222574 TI - Content validity of the Quebec licensing examination (OSCE). Assessed by practising physicians. AB - OBJECTIVE: To assess content validity of the objective structured clinical examination (OSCE), which uses multiple encounters with standardized patients and which is included in the Quebec licensing examination (QLEx). DESIGN: Descriptive study using written questionnaires based on Likert-type scales. SETTING: Examination centres of the QLEx. PARTICIPANTS: Thirteen practising family physicians with 3 to 6 years' practice experience who passed the QLEx OSCE in May 1993. MAIN OUTCOME MEASURES: Opinion whether the QLEx OSCE globally measured the competence of family physicians; opinion whether OSCE cases and stations made it possible to assess the main clinical abilities required of a family physician; opinion on specific statements about each case, such as how frequently the case was seen in the participants' practice, if the participants knew how to deal with the problem, if the simulated patient was believable, if the clinical situation was realistic, and if the duration of the station was adequate; and participants' self-assessment of their performance on the case. RESULTS: Eleven participants (84.6%) agreed that the OSCE globally measured the competence of a family physician. All agreed that the OSCE cases and stations made it possible to assess the main clinical skills required of a family physician. For most of the cases, participants believed that they knew how to deal with the problem, that the clinical situation was realistic, that the simulated patient was believable, and that the case duration was adequate. CONCLUSIONS: The results of this study support the content validity of the QLEx OSCE and confirm that it assesses the main skills needed for practising family medicine and adequately samples possible encounters or cases seen in family practice. PMID- 9222575 TI - Willingness to follow breast cancer. Survey of family physicians. AB - OBJECTIVE: To identify the experience and willingness of family physicians to accept follow-up care of patients treated for stage I breast cancer. DESIGN: Mailed questionnaire. PARTICIPANTS: One hundred eighty-nine family physicians in southwestern Ontario with oversampling of female physicians and physicians practising more than 20 km from a cancer clinic. MAIN OUTCOME MEASURE: Willingness to follow breast cancer patients and time after treatment family physicians would be willing to begin follow-up care. RESULTS: We had an 81.5% response rate. Of the 154 respondents, 53% had been involved previously in the 5 year, follow-up care of a patient with breast cancer and 77.1% believed it appropriate for family physicians to assume responsibility for follow-up care in all or most cases. If asked by a patient, the family, or an oncologist to provide follow-up care, 90.1% of family physicians reported they would accept this responsibility. Willingness to follow breast cancer patients was not associated with sex, years in practice, proximity to a cancer clinic, or certification status but was associated with having previously provided such care (P = .043). Of those willing to care for these patients, almost 90% would prefer to start within 1 year of treatment. CONCLUSION: Although only half the respondents had experience in providing follow-up care to breast cancer patients, most were willing to take on this role, especially if asked. PMID- 9222576 TI - Measuring the effectiveness of a pilot continuing medical education program. AB - OBJECTIVE: To evaluate a learner-centred, small group CME program intended to improve the clinical performance of family physicians identified as having serious practice deficiencies by the University of Manitoba's Clinical Assessment and Enhancement Program. DESIGN: Nonrandomized control trial in which data were collected from patients' charts and physician performance was evaluated. Differences in subjects' scores were tested at program entry and at 6 months and 18 months later using a two-way analysis of variance. SETTING: Family medicine practices in Manitoba. PARTICIPANTS: Fifteen family physicians: five study subjects and 10 control subjects. The five study subjects were identified as needing CME to improve their clinical performance. The 10 control subjects were randomly selected. INTERVENTIONS: Participants attended a 10-session, learner centred, small group CME program. MAIN OUTCOME MEASURES: Clinical care, preventive care, charting, and the use of drugs were the variables assessed. RESULTS: Study subjects' initial scores were much lower than those of controls, but improved significantly during the CME program. CONCLUSION: A learner-centred, small group CME program can improve clinical performance. PMID- 9222577 TI - Teaching home care to family medicine residents. AB - A growing elderly population suffering from chronic and debilitating diseases, the rising cost of institutional care, and increasing demand from patients for home visits indicate that home care will become a more important part of family physicians' practice in the future. We describe a model for teaching family medicine residents how to provide home services. PMID- 9222578 TI - Model for assessing psychosocial problems. AB - The Model for the Assessment of Psychosocial Problems (MAPP) can help family medicine residents effectively assess patients with psychosocial problems. Following a patient-centred clinical method, MAPP provides a guide to exploring problems and an approach that allows residents and patients jointly to define problems and decide upon management. Emphasis is placed on clarifying patients' expectations of physicians. PMID- 9222579 TI - Hospital grand rounds in family medicine. Content and educational structure. AB - OBJECTIVE: To investigate hospital grand rounds in family medicine, to examine their content and organization, and to recommend improved educational structures for these ubiquitous continuing medical education events. DATA SELECTION: Retrospective analysis of titles and content of 358 family medicine grand rounds offered in the department of family medicine of a large urban hospital from mid 1983 to the end of 1994. FINDINGS: Only 10% of family medicine grand rounds were presented by family physicians. Most grand rounds were in the form of specialists exhibiting their own interests in a lecture format. Analysis of grand rounds titles showed no consistent pattern of topics but an emphasis on practical aspects of medical care. Patient-based presentations were uncommon, as were grand rounds with more than one speaker. CONCLUSIONS: The content and mix of topics appeared appropriate, but in the absence of a curricular structure, or evaluation of learning gain, it is difficult to assess the value of grand rounds. PMID- 9222580 TI - Recognizing antiphospholipid syndrome. Case report: misdiagnosis delayed treatment. AB - Family physicians should be aware of antiphospholipid syndrome, a rare and recently described disorder, but one that can be treated successfully. Physicians should have a high degree of suspicion in patients with thrombotic symptoms if they are to detect and treat this condition early. PMID- 9222581 TI - Stress protein research in transplantation. Report on presentations at the Fourth Basic Sciences Symposium of the Transplantation Society. PMID- 9222582 TI - Chaperone function on Crete: a meeting report. PMID- 9222583 TI - Heat shock-induced myocardial protection against ischemic injury: a role for Hsp70? PMID- 9222584 TI - Heat shock proteins, molecular chaperones and the prion encephalopathies. PMID- 9222585 TI - A hitchhiker's guide to the human Hsp70 family. AB - The human Hsp70 family encompasses at least 11 genes which encode a group of highly related proteins. These proteins include both cognate and highly inducible members, at least some of which act as molecular chaperones. The location of cognate Hsp70s within all the major subcellular compartments is an indication of the importance of these proteins. The expression of several inducible Hsp70 genes is also an indication of the importance of these proteins in the stress response. The existence of multiple genes and protein isoforms has created confusion in the identification and naming of particular family members. We have compiled, from the literature, a list of genes and genetic loci and produced a two-dimensional protein map of the known human Hsp70 family members. This will enable researchers in the field to quickly and reliably identify human Hsp70s. We have also devised a more rational nomenclature for these genes and gene products which, subject to general acceptance, could be extended to Hsp70 families from other species. PMID- 9222586 TI - Review of patents in the cell stress and chaperone field. PMID- 9222587 TI - Evidence for a role of Hsp70 in the regulation of the heat shock response in mammalian cells. AB - Heat and other environmental insults (stress) cause unfolding of proteins, triggering the activation of heat shock transcription factor HSF (HSF1 in vertebrates) that, in higher eukaryotes, involves trimerization of the factor and acquisition of heat shock element (HSE) DNA-binding ability. Interaction of activated HSF1 with HSEs in promoters of genes encoding heat shock proteins (Hsps) enhances their expression. It was suggested that Hsp70 may function as the negative regulator of HSF1. In the simplest model, stress-unfolded proteins would compete with monomeric HSF1 for Hsp70 binding. This competition would result in dissociation of an HSF1-Hsp70 complex, allowing trimerization of released HSF1 monomers. In support of this model, we present evidence herein that 1) non activated HSF1 forms a 1:1 complex with Hsp70, 2) both rates of heat-induced appearance of HSF1 oligomers and rates of disappearance of HSF1 heterodimers and monomers decrease when concentrations of unengaged Hsps are increased, and 3) transient overexpression of Hsp70 inhibits heat activation of HSF1. PMID- 9222588 TI - Cell-specific heat-shock induction of Hsp23 in the eye of Drosophila melanogaster. AB - The expression of two small heat shock proteins (sHsp), Hsp23 and Hsp27, was examined by immunological approaches in the eye of Drosophila melanogaster. Neither Hsp23 nor Hsp27 is detectable in unstressed (23 degrees C) eyes but both proteins are induced by heat shock (35 degrees C). In response to heat stress, Hsp27 is expressed in all cells of the ommatidium including the cone, pigment and photoreceptor cells. However, the heat-induced expression of Hsp23 is restricted to a single cell type of the ommatidium, the cone cells, suggesting that Hsp23 is regulated by specific mechanisms acting to inhibit the expression of this polypeptide in some ommatidial cells. The cell-specific induction of Hsp23 under stress conditions does not seem to be regulated by the Drosophila melanogaster heat shock transcriptional factor (DmHSF). In both unstressed and stressed conditions, DmHSF is detected in all the different types of ommatidial cells where it is found associated with the nucleus. These observations suggest that factors, other than the heat shock transcriptional factor, are involved in regulating the expression of the hsp23 gene under stress conditions. PMID- 9222590 TI - Evaluation of stress-inducible hsp90 gene expression as a potential molecular biomarker in Xenopus laevis. AB - In this study we have evaluated stress-inducible hsp90 mRNA accumulation as a potential molecular biomarker in Xenopus laevis. In order to obtain a probe for Northern blot analysis we employed a PCR-based approach using degenerate primers for the amplification and cloning of an hsp90 gene sequence from Xenopus laevis. The deduced amino acid sequence is 102 amino acids in length and exhibited the highest degree of identity with zebrafish and human hsp90 beta genes. Furthermore, the putative intron and exon boundaries of this fragment are the same as hsp90 beta in chicken, mouse and human, indicating that the fragment represents a Xenopus hsp90 beta-like gene. Northern blot analyses revealed that this gene was constitutively expressed in cultured A6 cells. While heat shock and sodium arsenite exposure resulted in the increased accumulation of hsp90 mRNA in A6 cells, treatment with cadmium chloride and zinc chloride did not. Also, exposure of A6 cells to concurrent heat shock and sodium arsenite produced a mild synergistic response with respect to hsp90 mRNA levels in contrast to hsp70 mRNA levels which displayed a strong synergistic effect. Finally, hsp90 mRNA was detected constitutively throughout early embryogenesis but was heat-inducible only in late blastula and later stages of development. Given the normal abundance and limited stress-induced accumulation of hsp90 mRNA, it may not have a great deal of potential as a molecular biomarker compared to hsp70 and hsp30 mRNA. However, it may be useful in conjunction with other stress protein mRNAs to establish a set of biomarker profiles to characterize the cellular response to a stressful or toxic agent. PMID- 9222589 TI - Amplification and altered expression of the hsc70/U14 snoRNA gene in a heat resistant Chinese hamster cell line. AB - We have recently demonstrated that the heat resistant phenotype of the HR-1 variant isolated from HA-1 Chinese hamster fibroblasts after a series of heat shocks is associated with the increased expression of Hsc70, the constitutive form of Hsp70 (Laszlo and Li 1985). Here, we report the cloning and characterization of the Chinese hamster hsc70 gene and its organization and expression in wild type HA-1 and permanently heat resistant HR-1 cells. DNA sequencing revealed that the structure and nucleotide sequence of the hamster hsc70 gene is highly homologous to the human and rat genes coding for Hsc70. Three of the eight introns of the hamster hsc70 gene encode U14 small nucleolar RNAs, as has been demonstrated in other species. Although putative transcriptional elements, including a TATA box, two inverted CAT boxes, and two sets of heat shock elements (HSEs) are completely conserved in the human and hamster hsc70 genes, the regulation of expression of the hamster hsc70 gene is different from that reported for its human counterpart in that the mRNA coding for Hsc70 increases at least 10-fold after a mild heat shock in Chinese hamster cells while no induction of Hsc70 by heat shock has been reported in human cell lines. In situ hybridization revealed a complex chromosomal rearrangement in HR-1 cells which results in the 4- to 5-fold amplification of the hsc70 gene as indicated by genomic Southern blots. In association with this amplification of the hsc70 gene, the levels of Hsc70 mRNA and U14 snoRNA are increased in the HR-1 cells under both normal growing conditions and after heat shock. Thus, the elevated expression of both Hsc70 and U14 snoRNA might play a role in the heat resistant phenotype of the HR-1 cells. This is the first report of the amplification of a heat shock gene and the possible induction of gene amplification by heat shock. PMID- 9222591 TI - Translational thermotolerance in Saccharomyces cerevisiae. AB - While protein synthesis is rapidly inactivated in Saccharomyces cerevisiae, cells shifted from log growth at 30 degrees C to 43 degrees C, a 1-h 37 degrees C treatment given to cells just prior to the shift to 43 degrees C partially blocks this inactivation. By contrast, such a pre-heat shock treatment has no protective effect on translational inactivation at 45 degrees C or higher. Cells allowed to approach stationary phase not only develop an enhanced thermotolerance relative to log cells but also exhibit a pronounced resistance to inactivation of protein synthesis at 43 degrees C as well as at 45 degrees C. We have found that this 'translational thermotolerance' can also be induced in S. cerevisiae by briefly treating log phase cells at 30 degrees C with cycloheximide. Using such a procedure to induce stabilization of protein synthesis at 43 degrees C, we have been able to show that heat shock-induced proteins are not responsible for the establishment of this protective effect. This work shows that enhanced thermotolerance can be induced in log cells even after a shift to 43 degrees C, as long as a prior translational thermotolerance has been established. Furthermore, we show that the capacity of plateau cells to maintain translation at 43 degrees C contributes significantly to their state of enhanced thermotolerance. PMID- 9222592 TI - Thermal activation of the bovine Hsc70 molecular chaperone at physiological temperatures: physical evidence of a molecular thermometer. AB - Differential scanning calorimetry was used to monitor the thermal transitions of the 70 kDa heat shock cognate protein (Hsc70). Hsc70 had endothermic transitions with midpoints (Tm) at 59 degrees C and 63 degrees C in the absence and presence of ATP, respectively, and a similar increase in Tm was observed using intrinsic fluorescence of tryptophan. Combined with increased exposure at 60 degrees C of non-polar residues of Hsc70 to which the hydrophobic, fluorescent probe ANS bound, these data indicate that the endotherms represent thermal denaturation and that bound nucleotide stabilizes Hsc70. An exothermic transition (Tm = 66 degrees C) was detected by calorimetry for Hsc70-apocytochrome c (apo c) complexes. An increase in intrinsic fluorescence with the same Tm and increased turbidity indicated aggregation of the denatured Hsc70-apo c. A novel finding was an exothermic transition of Hsc70 beginning at about 30 degrees C (Tm = 41 degrees C). No changes in either intrinsic fluorescence or ANS fluorescence attributable to protein transitions were detected in this temperature range. Examination of samples run on native polyacrylamide gels indicated that this exothermic transition was not due to Hsc70 aggregation or multimer formation. However, Hsc70 was protease-resistant at 20 degrees C, sensitive at 40 degrees C and resistant when returned to 20 degrees C, indicating that this exotherm is associated with a reversible conformational change. As an assay for Hsc70 chaperoning function, complex formation was measured as a function of temperature using a variety of substrates including the model unfolded protein apo c, a pigeon cytochrome c fragment, a representative hydrophobic-aromatic peptide FYQLALT, and a representative hydrophobic-basic motif NIVRKKK. For all of these substrates, the amount of complex formed increased with increasing temperature over the same range as the 41 degrees C exotherm. It is proposed that a conformational change exposes polar and charged residues in Hsc70 which subsequently become hydrated, resulting in an active chaperone. Hsc70 may be a thermal sensor that matches the supply of chaperoning activity with demand for it over the physiological temperature range of mammalian cells. Thermal activation of Hsc70 may also have a role in acquired thermotolerance. PMID- 9222593 TI - Recent literature on cell stress & chaperones. PMID- 9222594 TI - Discovery of the heat shock response. PMID- 9222595 TI - Stress proteins as agents of immunological change: some lessons from metallothionein. AB - The stress response proteins each have somewhat unique characteristics that enable them to function under conditions of cellular stress, and to contribute to cellular survival in difficult times. The immune response is, by definition, a mechanism that often operates in times of cellular stress, and even creates stress during its operation. Cells called upon to respond to tissue damage caused by inflammation can have extraordinary demands placed upon them and surrounding tissue may suffer damaging conditions that were originally established to eliminate the source of the inflammation. Stress proteins may be released from some of these damaged cells as a programmed response to the stress, or as a simple consequence of excessive damage to the plasma membrane. In either instance, there is the opportunity for these stress proteins to interact with cells and proteins in the extracellular environment. It may be that those same characteristics that enable stress proteins to interact with structures within the cell also enable interactions outside the cell, but with dramatically different results. As has been found with MT, interference with these extracellular interactions may decrease the consequences of stress on the immune response, and may enable more effective immunity. It may also be possible to employ the various stress proteins to manipulate normal immune function. PMID- 9222596 TI - Influence of molecular and chemical chaperones on protein folding. AB - Protein folding inside the cell involves the participation of accessory components known as molecular chaperones. In addition to their active participation in the folding process, molecular chaperones serve as a type of 'quality control system', recognizing, retaining and targeting misfolded proteins for their eventual degradation. It is now known that a number of human diseases arise as a consequence of specific point mutations or deletions within genes encoding essential proteins. In many cases these mutations/deletions are not so severe as to totally destroy the biological activity of the particular gene product. Rather, the mutations often result in only subtle folding abnormalities which lead to the newly synthesized protein being retained at the endoplasmic reticulum by the actions of the cellular quality control system. In this short review article we discuss our recent studies showing that the protein folding defect associated with the most common mutation in patients with cystic fibrosis can be overcome by a novel strategy. As shown in the paper by Brown et al in this issue (Brown et al 1996), a number of different low molecular weight compounds, all known to stabilize proteins in their native conformation, are effective in rescuing the processing defect of the mutant cystic fibrosis transmembrane conductance regulator protein. We then discuss how these same compounds, which we now call chemical chaperones, also may prove to be effective in correcting a number of other protein folding abnormalities which constitute the underlying basis of a large number of different human diseases. PMID- 9222597 TI - Chemical chaperones correct the mutant phenotype of the delta F508 cystic fibrosis transmembrane conductance regulator protein. AB - Mutations in the cystic fibrosis transmembrane conductance regulator protein (CFTR) often result in a failure of the protein to be properly processed at the level of the endoplasmic reticulum (ER) and subsequently transported to the plasma membrane. The folding defect associated with the most common CFTR mutation (delta F508) has been shown to be temperature sensitive. Incubation of cells expressing delta F508 CFTR at lower growth temperatures results in the proper processing of a portion of the mutant CFTR protein. Under these conditions, the mutant protein can move to the plasma membrane where it functions, similar to the wild-type protein, in mediating chloride transport. We set out to identify other methods, which like temperature treatment, would rescue the folding defect associated with the delta F508 CFTR mutation. Here we show that treatment of cells expressing the delta F508 mutant with a number of low molecular weight compounds, all known to stabilize proteins in their native conformation, results in the correct processing of the mutant CFTR protein and its deposition at the plasma membrane. Such compounds included the cellular osmolytes glycerol and trimethylamine N-oxide, as well as deuterated water. Treatment of the delta F508 CFTR-expressing cells with any one of these compounds, which we now refer to as 'chemical chaperones', restored the ability of the mutant cells to exhibit forskolin-dependent chloride transport, similar to that observed for the cells expressing the wild-type CFTR protein. We suggest that the use of 'chemical chaperones' may prove to be effective for the treatment of cystic fibrosis, as well as other genetic diseases whose underlying basis involves defective protein folding and/or a failure in normal protein trafficking events. PMID- 9222598 TI - Physical interactions between members of the DnaK chaperone machinery: characterization of the DnaK.GrpE complex. AB - Many of the functions of the Escherichia coli Hsp70, Dnak, require two cofactors, DnaJ and GrpE. GrpE acts as a nucleotide exchange factor in the DnaK reaction cycle but the details of its mechanism remain unclear. GrpE has high affinity for monomeric native DnaK, with a Kd estimated at < or = 50 nM. GrpE is a very asymmetric molecule and exists as either a dimer or trimer in its native state. The stoichiometry of GrpE to DnaK in the isolated complex was 3:1, suggesting a trimer. Formation of the complex is quite fast (kan > 1 s-1), whereas the off rate is very slow on the HPLC timescale (kant < or = 10(-4) s-1). GrpE has no affinity for ATP or ADP, nor the oligomeric and molten globule states of DnaK. The complex is much more thermally stable than either GrpE or DnaK alone, and prevents the formation of the molten globule-like state of DnaK at physiologically relevant temperatures. Formation of the complex does not cause any change in secondary structure, as determined by the lack of change in the circular dichroism spectrum. However, binding of GrpE induces a similar tertiary structural change in DnaK to that induced by binding of ATP, based on the blue shift in lambda max from the fluorescence of the single tryptophan in DnaK. The nucleotide exchange properties of GrpE can be explained by the conformational change which may represent the opening of the nucleotide cleft on DnaK, subsequently inducing a low affinity state for ADP. PMID- 9222599 TI - Quantitative evidence that both Hsc70 and Hsp70 contribute to thermal adaptation in hybrids of the livebearing fishes Poeciliopsis. AB - The 70-kilodalton heat shock protein family is composed of both environmentally inducible (Hsp) and constitutively expressed (Hsc) family members. While the role of the constitutively expressed stress proteins in thermotolerance is largely unknown, de novo expression of stress proteins in response to elevated temperatures has been associated with increased thermotolerance in many cell lines, developing embryos and adult organisms. Distinct, hemiclonal hybrids between the livebearing fish species Poeciliopsis monacha and P. lucida varied in their abilities to survive temperature stress, with survival being greatest when rates of temperature increase to 40 degrees C were slowest and when P. monacha genomes were combined with a sympatric P. lucida genome. Quantification of Hsp70 under heat shock conditions and Hsc70 under normal physiological conditions indicated that variation in survival among hemiclones was best explained by the combined effects of these two proteins. Similar complex interactions between maternal and paternal genomes and rate of temperature increase were found to underlie patterns of survival, Hsp70 accumulation and Hsc70 abundance. These data suggest that the relationship between Hsps and thermotolerance is more intricate than previously thought and that Hsps contribute to thermal adaptation in these fishes through genetic interactions specific to particular environments. PMID- 9222600 TI - Discovery of molecular chaperones. PMID- 9222601 TI - Exogenous heat shock cognate protein Hsc 70 prevents axotomy-induced death of spinal sensory neurons. AB - Elevation of intracellular heat shock protein (Hsp)70 increases resistance of cells to many physical and metabolic insults. We tested the hypothesis that treatment with Hsc70 can also produce that effect, using the model of axotomy induced neuronal death in the neonatal mouse. The sciatic nerve was sectioned and in some animals purified bovine brain Hsc70 was applied to the proximal end of the nerve immediately thereafter and again 3 days later. Seven days postaxotomy, the surviving sensory neurons of the lumbar dorsal root ganglion (DRG) and motoneurons of the lumbar ventral spinal cord were counted to assess cell death. Axotomy induced the death of approximately 33% of DRG neurons and 50% of motoneurons, when examined 7 days postinjury. Application of exogenous Hsc70 prevented axotomy-induced death of virtually all sensory neurons, but did not significantly alter motoneuron death. Thus, Hsc70 may prove to be useful in the repair of peripheral sensory nerve damage. PMID- 9222602 TI - Cell surface expression of heat shock proteins and the immune response. PMID- 9222604 TI - Increased HSF activation in muscles with a high constitutive Hsp70 expression. AB - Stress-induced transcriptional regulation of the Hsps is mediated by trimerization and binding of a pre-existing heat shock transcription factor (HSF1) to a specific DNA sequence located in the 5' region of hsp genes, known as the heat shock element. Hsp70 has been implicated in regulating the activation of the HSF and, according to cell culture models, high steady-state levels of Hsp70 are inversely correlated with HSF activation. To determine if this applies in an intact animal, muscles of the rat hindlimb which differ in the constitutive expression of Hsp70, were assessed for HSF activation following heat shock. Mobility shift gel analyses demonstrated that HSF activation was detectable in extracts from all muscles following heat shock regardless of Hsp70 content. However, muscles comprised predominantly of slow/Type I fibers (soleus) demonstrated a greater HSF activation, as well as a faster HSF activation and inactivation, than muscles comprised predominantly of fast/Type II fibers (white gastrocnemius). In addition, muscles pretreated by two heat shocks (24 h apart) demonstrated a stronger HSF activation than muscles subjected to only one heat shock. Thus, results from cell culture models demonstrating that tissue levels of Hsp70 are inversely correlated with HSF activation, may not apply to the muscles of an intact animal. PMID- 9222603 TI - Identification of the C-terminal region of 70 kDa heat shock protein from Leishmania (Viannia) braziliensis as a target for the humoral immune response. AB - A Leishmania (Viannia) braziliensis (Lb) promastigote cDNA library in lambda gt11 was screened with patients' sera with the aim of identifying antigens specifically related to mucocutaneous leishmaniasis (MCL). One of the clones isolated, 133P, consistently reacted with MCL sera; it was sequenced and found to encode the C-terminal three-quarters of a protein belonging to the highly conserved Hsp70 family. Since Hsp70 proteins from different species tend to be less conserved through the C-terminal end, it was predicted that this region would be more antigenic and was likely to bear the discriminatory epitopes. In order to test this hypothesis, the N- and C-terminal halves of the polypeptide encoded by 133P, 133P-N and 133P-C, respectively, were expressed in Escherichia coli. Immunoblotting analysis demonstrated that 133P-C reacted more strongly with a pool of MCL sera than 133P-N, and both recombinant proteins reacted faintly with pools of cutaneous (CL) and visceral (VL) leishmaniasis sera. These results confirmed the predicted epitope location in the C-terminal region. The 133P-C fragment was also expressed as a fusion protein with glutathione-S-transferase (GST-133P-C), affinity-purified with glutathione-agarose and tested by ELISA with individual sera. From 46 Lb-infected patients, 41 sera (89%) were positive, no cross-reactivity was observed with healthy, Trypanosoma cruzior L. amazonensis infected individuals. Despite a relatively high cross-reactivity with VL sera, the enhanced humoral response of MCL as compared with CL patients might be interesting for studies on disease aggravation. PMID- 9222605 TI - Enhancement of glucocorticoid receptor-mediated gene expression by cellular stress: evidence for the involvement of a heat shock-initiated factor or process during recovery from stress. AB - Recent reports have demonstrated the ability of cellular stress to cause a large increase in the maximal levels of steroid receptor-mediated gene expression, a process we refer to as the heat shock potentiation effect (HSPE). In the present work, we have analyzed the time of appearance of the HSPE on the glucocorticoid receptor (GR) of L929 cells stably-transfected with the MMTV-CAT reporter plasmid (LMCAT2 cells). In LMCAT2 cells exposed to heat shock (43 degrees C, 2-h) before addition of 1 microM dexamethasone, the first appearance of HSPE (CAT levels greater than hormone-alone) occurred at 8 h of recovery and continued to increase by 24 h of recovery. Treatment of LMCAT2 cells with 1 microM dexamethasone for 2 h before heat or chemical shock (sodium arsenite) resulted in the same delayed onset pattern for the HSPE. Based on a [35S]methionine assay and tests of L929 cells stably transfected with the constitutive pSV2-CAT reporter, evidence is provided that the delayed appearance of the HSPE is not due to the heat shock block of general protein synthesis or to specific repression of CAT mRNA expression or translation. By using short-term incubations (4 h) with dexamethasone during the recovery period, the peaks of HSPE expression during recovery were determined to be 12-16 h for CAT enzyme activities, and 4-8 h for CAT mRNA expression. Taken together, these results provide evidence that the timing of the HSPE is not dependent on the rate of GR activation, or on the type of stress, but rather on a factor or process that is either synthesized or activated during the recovery period following stress. PMID- 9222606 TI - Heat shock in vertebrate cells. PMID- 9222607 TI - The Hsf world: classification and properties of plant heat stress transcription factors. AB - Based on the partial or complete sequences of 14 plant heat stress transcription factors (Hsfs) from tomato, soybean, Arabidopsis and maize we propose a general nomenclature with two basic classes, i.e. classes A and B each containing two or more types of Hsfs (HsfA1, HsfA2 etc.). Despite some plant-specific peculiarities, essential functional domains and modules of these proteins are conserved among plants, yeast, Drosophila and vertebrates. A revised terminology of these parts follows recommendations agreed upon among the authors and representatives from other laboratories working in this field (see legend to Fig. 1). Similar to the situation with the small heat shock proteins (sHsps), the complexity of the hsf gene family in plants appears to be higher than in other eukaryotic organisms. PMID- 9222608 TI - Transient accumulation, phosphorylation and changes in the oligomerization of Hsp27 during retinoic acid-induced differentiation of HL-60 cells: possible role in the control of cellular growth and differentiation. AB - Expression of the mammalian small stress protein Hsp27 has been increasingly linked to cell growth regulation and differentiation. Hsp27 is a phosphoprotein which forms oligomers with native sizes ranging between 100 and 800 kDa. We have examined the fate of Hsp27 transiently expressed during the retinoic acid (tRA) induced granulocytic differentiation of human leukemic HL-60 cells. We show that tRA, in addition to its effects on Hsp27 accumulation and phosphorylation, transiently increased the oligomerization state of this protein. While Hsp27 phosphorylation by tRA is an early phenomenon that takes place before cellular growth is altered, the redistribution of Hsp27 oligomers occurred later and concomitantly with the maximal accumulation of this protein. Hence, complex regulations of Hsp27 are induced by tRA which suggest that this protein plays a role in the pathway through which retinoids exert their effects. To approach Hsp27 function during HL-60 cell differentiation, experiments aimed at reducing the cellular content of this protein were performed by transiently inhibiting Hsp27 mRNA translation by a specific anti-sense oligonucleotide. This process, which decreased the basal level of Hsp27 by about 40%, did not interfere with the growth of undifferentiated HL-60 cells. In contrast, a decreased level of Hsp27 diminished the differentiation-mediated down-regulation of cell growth and altered some morphological changes induced by this retinoid. These results suggest that Hsp27 is a mediator of granulocytic differentiation. PMID- 9222609 TI - A pathway of multi-chaperone interactions common to diverse regulatory proteins: estrogen receptor, Fes tyrosine kinase, heat shock transcription factor Hsf1, and the aryl hydrocarbon receptor. AB - A variety of regulatory proteins, including different classes of transcription factors and protein kinases, have been identified in complexes with Hsp90. On careful examination of unactivated progesterone receptor complexes, eight different protein participants have been identified, and each can be considered a component of the cytoplasmic molecular chaperone machinery. These proteins are Hsp90, Hsp70, Hip, p60, p23, FKBP51, FKBP52 and Cyp40. Studies in a cell-free assembly system have helped to define a highly ordered, dynamic pathway for assembly of progesterone receptor complexes. In the present study, target proteins other than progesterone receptor were used in this cell-free system to assemble complexes in vitro and to compare the composition of resulting complexes. Targets used were human estrogen receptor, human Fes protein-tyrosine kinase, human heat shock transcription factor Hsf1, and human aryl hydrocarbon receptor. The striking similarity of resulting target complexes with previously characterized progesterone receptor complexes suggest that each of these targets undergoes a common assembly pathway involving multiple chaperone components in addition to Hsp90. PMID- 9222610 TI - Diminished heat shock response in the aged myocardium. AB - Induction of heat shock proteins (Hsps), Hsp72 in particular, has been associated with myocardial protection. Since a decreased Hsp response has been reported to occur with aging, it was of interest to determine if hearts from aged animals also demonstrate an altered heat shock response and subsequent myocardial protection. Adult (6 months old) and aged (22 months old) Fischer 344 rats were heat stressed by raising their rectal temperatures to 41 degrees C for 10 min. At selected times following heat stress (0-24 h) hearts were examined for heat shock transcription factor trimerization and DNA-binding activity (Hsf1 activation), Hsp72 mRNA accumulation, Hsp72 and Hsf1 protein content, as well as, protection from ischemia using the Langendorff isolated heart model. Following heat stress, hearts from aged animals demonstrated a 47% reduction in Hsf1 activation, a reduction in Hsp72 mRNA and a 35% reduction in Hsp72 protein content, compared to hearts from adults. Interestingly, myocardial Hsf1 protein content was similar between aged and adult animals. Hearts from heat stressed adult animals (24-h prior) demonstrated an enhanced postischemic recovery as indicated by a greater recovery of left ventricular pressure and rate of contraction (P < 0.05), while hearts from heat stressed aged animals failed to demonstrate an enhanced postischemic recovery. These results suggest that hearts from aged animals exhibit an impaired ability to produce the protective Hsps and thus, may explain, at least in part, the increased susceptibility of aged hearts to stress. PMID- 9222611 TI - Quantitative vs qualitative methods. PMID- 9222613 TI - Incidence and nature of feeding problems in young children referred to a paediatric HIV service in London: FEAD screening. AB - A detailed screening assessment was carried out on two matched groups of young children; one group was HIV-infected and the other was not. Screening included assessments of growth, development and food intake. Parents were also interviewed about their child's feeding and mealtime behaviours. Half of the HIV-infected children were reported with serious feeding problems; significantly higher than in the uninfected group. More of the children in the HIV-infected group were found to have poorer growth and developmental weaknesses than in the uninfected group. A combination of physical and psychological factors are suggested as contributing to these feeding difficulties. Early monitoring of feeding behaviours, daily routines and food intake, together with systematic growth and developmental measures are suggested as important components in the care and management of HIV-infected children. PMID- 9222612 TI - Assessing social work effectiveness in child care practice: the contribution of randomized controlled trials. AB - This article discusses the role of randomized controlled trials (RCTs) in evaluating the impact of social work interventions with children. While recognizing the difficulties of applying RCTs to all aspects of practice, we argue that controlled trials can provide the most convincing evidence of the impact of social work activities on the welfare of children and families. Accumulating evidence of the effectiveness of interventions, we propose, should constitute the core business of social work research. To this end, it is necessary to recognize the primacy of the randomized controlled trial in exploring the relationship between social work activities and client outcomes. PMID- 9222614 TI - Motor difficulties in children with developmental disorders of speech and language. AB - The motor hand function of 16 children, aged between 4 and 7 years, with developmental speech and language disorders, was compared with that of 16 control children. The children with developmental speech and language disorders were significantly slower than controls on three out of four motor tasks. They were also more likely than controls to have mixed hand preference although this results was not significant. Children with developmental speech and language disorders should be assessed to ensure that motor deficits are diagnosed and that appropriate support is given. PMID- 9222615 TI - Child Health Surveillance in England and Wales: the good news. AB - Postal surveys were conducted in 1993 among all, or samples of, six groups of providers and managers of pre-school child health surveillance (CHS) in England and Wales. Content analyses were also carried out of strategic policy statements for CHS produced by 54 district health authorities in England and Wales. The surveys aimed to document the views and experiences of CHS providers and managers about the impact of recent changes affecting the structure and operation of CHS, including the publication of Health for All Children, the 1990 Contract for General Practitioners (GPs), the implementation of the National Health Service and Community Care Act 1990, and the changing roles of community doctors and health visitors. Five positive findings from the surveys are discussed: the impact of the first edition of Health for All Children; improvements in the development and use of child health information systems; the beneficial effects of the growing Involvement of GPs in CHS; the developing understanding of, and commitment to, the principle of clinical audit in CHS; and the growing collaboration between providers in the NHS internal market. A separate paper reports the negative findings from the study. PMID- 9222616 TI - Child Health Surveillance in England and Wales: the bad news. AB - Postal surveys were conducted in 1993 among all, or samples of, six groups of providers and managers of pre-school child health surveillance (CHS) in England and Wales. Content analyses were also carried out of strategic policy statements for CHS produced by 54 district health authorities in England and Wales. The surveys aimed to document the views and experiences of CHS providers and managers about the impact of recent changes affecting the structure and operation of CHS, including the publication of Health for All Children, the 1990 Contract for General Practitioners (GPs), the implementation of the National Health Service and Community Care Act 1990, and the changing roles of community doctors and health visitors. Four adverse findings from the surveys are discussed: fragmentation in the child health service; the unwanted effects of the NHS internal market; the adverse consequences of the changing role of health visitors; and the concerns voiced by the community doctors about the quality of CHS in general practice. PMID- 9222617 TI - Even young children's assessments provide useful information about asthma control. PMID- 9222619 TI - Acoustic rhinometry: its place in rhinology. PMID- 9222618 TI - Injuries and the risk of disability in teenagers and young adults. PMID- 9222620 TI - Whiplash injury. A clinical review with emphasis on neuro-otological aspects. PMID- 9222621 TI - Care of the professional voice. PMID- 9222622 TI - Parotidectomy through a rhytidectomy incision. PMID- 9222623 TI - A prospective study of PET-FDG imaging for the assessment of head and neck squamous cell carcinoma. AB - The main aim of the study was to evaluate the use of positron emission tomography using fluoro-deoxyglucose (PET-FDG) imaging for the detection of squamous cell carcinoma of the head and neck. Fifty-four consecutive patients with malignancies involving the head and neck were studied prospectively. Thirty-one patients presented with primary disease and 23 were suspected of recurrent or residual disease. All patients underwent full clinical staging, PET-FDG scans and anatomical imaging, 37 underwent computed tomography (CT), 13 magnetic resonance (MR) and four had both CT and MR. Clinical assessment, CT/MR, PET-FDG and histological examination were all evaluated independently of each other. All 31 primary head and neck malignant tumours were detected by PET-FDG. Based on 16 patients who underwent neck dissections, the sensitivity and specificity of PET FDG for detecting nodal disease was 67% and 100% respectively, compared with clinical assessment of 58% and 75% and CT/MR of 67% and 25%. In all 12 patients, PET-FDG correctly identified the presence of absence or recurrent or residual disease. PET-FDG staged 13 post-treatment necks with an accuracy of 100% as compared to CT/MR which was accurate in 7 of 13 and clinical assessment which was accurate in eight. Three sites of abnormal tracer uptake unrelated to malignancy were recorded as incidental findings (mandibular osteomyelitis, 1: post glossectomy site, 2). PET-FDG was more accurate than CT/MR for identifying primary and recurrent tumours as well as metastatic lesions in the neck. If these diagnostic properties of PET-FDG are confirmed in further prospective studies, it could prove a valuable adjunct for the management of head and neck cancer. PMID- 9222624 TI - Suitability of children for ENT day case procedures. AB - This prospective study evaluates the suitability of 500 children under 12 years of age using both medical and social criteria, to determine the proportion of children suitable for increasing the scope of ENT operations performed as day cases. Co-existing medical conditions that could interfere with both surgery and anaesthesia were found in 39 (8%). A combination of adverse social and domestic conditions, including the number of adults and children resident, number of bedrooms and individuals sleeping in the child's bedroom, the number of cars, and the presence of a telephone, were present in 171 (34%) of children's homes and were almost exclusively found in National Health Service (NHS) patients. Private patients, 31 (6%) were more suitable for day surgery. Guidelines need to be followed to minimize the risk if the number of procedures is increased, if more patients were treated privately, the proportionate number of patients requiring in patient care would rise in the National Health Service (NHS). PMID- 9222625 TI - The nasal response to axillary pressure in non-eosinophilic intrinsic rhinitis. AB - The exact pathophysiology of intrinsic rhinitis is not fully understood. The generally held belief is that it is due to an imbalance between the outflow of the sympathetic and parasympathetic nervous systems to the nose, perhaps due to excessive parasympathetic or reduced sympathetic activity. In this study the nasal airway response to a predominantly sympathetic stimulus, axillary pressure was studied in 19 patients with intrinsic rhinitis and compared with 16 normal patients. Axillary pressure was applied using a crutch. Following sustained pressure, a significant fall in nasal resistance in the normal group (0.823 kPas/l) and an insignificant fall in the patients with rhinitis (0.0725 kPas/l) was found. Pulse and blood pressure changes were similar in both groups with a significant rise in pulse rate and diastolic blood pressure. The study shows that there is an abnormal response to axillary pressure in intrinsic rhinitis, perhaps due to relative nasal sympathetic hyposensitivity. PMID- 9222627 TI - Aesthetic sequelae of septoplasty. AB - This prospective study, using standardized pre- and post-operative photographs examined by three independent observers, included 100 septoplasty patients with a minimal follow up of 9 months. A risk of minimal aesthetic changes (21%) could be documented, but significant post-operative changes (1%) were rare. Surprisingly, statistical evaluation of operative data in relation to aesthetic changes could not identify specific surgical manoeuvres which may increase the risk of aesthetic changes. All patients must be fully informed about possible changes in nasal shape as a result of septoplasty. Pre-operative photographs should be considered a prerequisite before any type of septal surgery. PMID- 9222626 TI - Auditory function in young patients with chronic renal failure. AB - Debate on the relationship between renal insufficiency and hearing loss continues mainly due to the advanced age and the possible accelerated presbycusis of the patients that have been studied in surveys. Hearing acuity was studied in 46 children and adolescents suffering from renal insufficiency. Sensorineural hearing loss (mainly high-frequency) of unknown cause was found in 14 patients (30.4%). Hearing loss was not influenced by the various haematological, biochemical and clinical parameter (type of renal disease, blood pressure, history of ototoxic drugs administration). However, hearing loss seemed to be affected by the method of management of the renal insufficiency (more in the haemodialysis group than in the peritoneal dialysis group). There were no significant changes in audiometric findings before and after haemodialysis. PMID- 9222628 TI - Glue ear surgery in Scottish children 1990-1994: still plenty of ENT and public health challenges. AB - This paper examines recent trends in glue ear surgery in Scottish children between 1990 and 1994, using routine National Health Service (NHS) data from all 15 Scottish Health Boards (total population 5,132,400 in 1994, with 1,038,296 aged 0-15). Absolute numbers, rates of glue ear operations, and variation in rates declined between 1990 and 1994 across all Health Boards. The proportion of glue ear operations which included grommet insertion increased. Grommet rates in children declined in those Boards with the highest rates, but increased in Boards with the lowest rates, thereby decreasing the variation in grommet rates across Scottish Health Boards from 3.8- in 1990 to 2.6-fold in 1994. Other operations for glue ear, particularly 'myringotomy and adenoidectomy', varied 20-fold between Health Boards. The proportion of operations performed as day cases increased, but day cases and repeat grommet insertions still showed two-fold variation across Scotland in 1994. Many aspects of surgical management still show variation, and merit further examination by ENT surgeons and Public Health physicians. PMID- 9222629 TI - Mandibular invasion in patients with oral and oropharyngeal squamous carcinoma. AB - Invasion of the mandible in squamous carcinoma of the oral cavity and oropharynx has always proved a problem for head and neck oncologists. We studied 82 patients who had mandibulectomies as part of their primary surgical treatment for cancer of these sites. In 40 patients, the tumour appeared to be invading the mandible on clinical grounds and 33 patients had tumours invading the mandible when the latter was examined histopathologically. Multivariate analysis showed that tumour was more likely to be fixed to and clinically invading the mandible in the presence of cancer of the oral cavity, compared with oropharyngeal cancer (P < 0.0001). There was a high degree of correlation between clinical invasion of the mandible and histopathological invasion of the mandible (P = 0.0059). In addition, clinical invasion of the mandible correlated with radiological findings (P = 0.0284). The 5-year survival of those patients with tumour that appeared not to be invading the mandible was 53% compared with 25% for those where tumour did appear to be invading the mandible (P < 0.02). The sensitivity and specificity of clinical evidence of mandibular invasion was calculated with the final arbiter of invasion being the histopathological findings. The sensitivity of clinical examination was 91% and the specificity 80%. The positive predictive value was 75% and the negative predictive value 93%. Mandibular invasion is a poor prognostic sign in cancer of the oropharynx and oral cavity. Detection of invasion prior to operation is obviously extremely important and it appears that clinical findings are an accurate method of predicting invasion. PMID- 9222630 TI - Tumours of the posterior pharyngeal wall: the use of the platysma flap. AB - Posterior pharyngeal will tumours are infrequent neoplasms with a very poor prognosis. The 5 year survival rate range from 3% to 32%. Most authors agree that the treatment of choice is surgery with post-operative radiotherapy. The results of treatment of 36 patients (tumour excision plus bilateral neck dissection and post-operative radiotherapy) in which the posterior pharyngeal wall defect was closed with a platysma myocutaneous flap were compared with other forms of repair (13 patients). The 5 year survival rate was 17.2% in the whole group. Laryngeal voice was achieved in 79% of patients having a platysma flap reconstruction. The platysma myocutaneous flap is very satisfactory for the repair of the posterior pharyngeal wall as it is easy to perform, it is oncologically safe and its functional results match well with other forms of reconstruction, with the advantage of laryngeal preservation. PMID- 9222631 TI - Preservation of the great auricular nerve during parotidectomy. AB - The great auricular nerve is routinely divided during the operation of parotidectomy, however, some surgeons have suggested that preserving the posterior branches reduces the area of post-operative anaesthesia. A prospective study was performed comparing the area of anaesthesia and hypoaesthesia in patients undergoing parotidectomy. In 20 patients the great auricular nerve was preserved and in 11 it was sacrificed. Mapping of the area of sensory loss at 2 weeks, 3, 6, 9 and 12 months showed that there was no difference between the two groups. The area of sensory loss decreased in an exponential fashion in both groups. The majority of the change occurred within 6 months. We conclude that preservation of the posterior branches of the great auricular nerve is unnecessary. PMID- 9222632 TI - The use of qualitative questionnaires in patients having and being considered for cochlear implants. AB - Within this study we have examined the particular hearing complaints of patients being considered for a cochlear implant and the specific benefits/shortcomings experienced by implanted patients using open-ended questionnaires. As we expected, the difficulty of hearing general conversation was the most common individual hearing complaint. However, 45% of complaints were concentrated in the 'psychosocial' category, which was significantly higher than that found among general audiological rehabilitation patients. The average number of benefits listed by patients having implants was significantly higher than that of the shortcomings. Moreover, the main benefits listed were focused on the acoustical and psychological factors, e.g. 'environmental sound awareness', 'general conversation easier' and 'feeling of self-confidence'. The main shortcomings were related to the acoustical and practical areas, e.g. hearing difficulty in noisy background, processor being cumbersome. PMID- 9222633 TI - Sensation of nasal airflow compared with nasal airway resistance in patients with rhinitis. AB - Over the past few decades there has been some controversy over the relationship between subjective assessment and objective measurement of nasal airway obstruction. To study the hypothesis that there is a close relationship between the two parameters, we analysed changes in nasal patency following histamine challenge. One hundred and two subjects with a history of allergic or non allergic rhinitis assessed their nasal patency on a visual analogue scale during nasal histamine provocation. Active anterior rhinomanometry was performed immediately after each patient assessment. At all points, significant correlations were observed between subjective and objective assessments of nasal obstruction. Regression analysis also provided strong evidence of a close relationship between the two parameters. We conclude that rhinomanometry can be used as an objective tool in determining nasal patency. PMID- 9222634 TI - Ludwig's angina. AB - A 13 year review of patients diagnosed to have Ludwig's angina admitted to the Christian Medical College and Hospital, Vellore, India, between March 1982 and April 1995 is presented. The patients were either admitted to the ENT or paediatric surgical units. There were 41 patients, 24% being children and 76% adults. The clinical profile and outcome of these two groups were compared. In the paediatric group, none had dental caries while in the adult group, 52% had associated dental caries. In children, 70% were controlled with conservative medical management unlike the adults of whom 81% needed incision and drainage. Tracheostomy was necessary in 10% of the children and in 52% of the adults. The mortality was 10% in both groups. PMID- 9222635 TI - Otological manifestations of primary ciliary dyskinesia. AB - Primary ciliary dyskinesia is a hereditary defect in the ultrastructure of cilia, leading to poor ciliary motility. The sinonasal and the bronchial manifestations of the disease are well documented; whereas its otological aspects have received less attention. In this report, we describe the clinical profile of 16 patients with primary ciliary dyskinesia laying particular emphasis on the otological manifestations. All children (11 patients) had bilateral otitis media with effusion. Of the five adults, three had tympanosclerosis; one had bilateral cholesteatoma; and one patient had bilateral keratosis obturans in combination with tympanosclerosis. Hearing improvement and a dry ear was achieved in all the children treated by tympanostomy tube insertion. The study suggests that otitis media is a prominent feature of this disorder. Most subjects suffer from protracted bilateral otitis media with effusion throughout childhood. PMID- 9222636 TI - Necrotizing external otitis: aminoglycoside and beta-lactam antibiotic treatment combined with surgical treatment. AB - Necrotizing external otitis is a life-threatening condition that still causes therapeutic problems. A retrospective analysis of 22 patients who were all treated with a short standard course of aminoglycoside and beta-lactam antibiotic with through local debridement was carried out; 50% (11/22) had diabetes mellitus, and all had a positive culture of Pseudomonas aeruginosa. The frequency of recurrence was 14% (3/22), and 95% (21/22) were eventually cured. The treatment course lasted 17 days on average. Side effects caused by the drug treatment were seen in 14% (3/22), but they were mild and transient consisting of a reversible increase of the serum creatinine level. It was concluded that the treatment was short, efficient, safe, and as successful as treatment with quinolones or third-generation cephalosporins which has been reported during the past years. In our opinion, therefore, these drugs should be reserved for cases of treatment failure, development of resistance, or side effects. PMID- 9222637 TI - Otomycosis: the detection of fungi in ears by immunofluorescence microscopy. AB - The procedure currently used to diagnose infection in otitis externa has several limitations: it is slow to culture organisms on growth media, fungal infections are often missed, and extensive laboratory facilities and mycological expertise are required. A rapid, accurate and sensitive assay would greatly improve patient care by initiating appropriate antifungal treatment at the onset of disease. We report the development of a rapid detection assay for otomycosis using fungal specific monoclonal antibodies to detect fungi in ear swabs by immunofluorescence microscopy. This assay could form the basis of a detection assay for fungal infections of the head and neck. PMID- 9222638 TI - The surgical management of drooling--a 15 year follow-up. AB - Drooling is rarely seen in the normal child after the age of 6 months, but an estimated 10% of children with neurological impairment suffer significant interference with everyday living due to excessive drooling. Submandibular duct relocation is a procedure that involves the dissection and re-routing of the submandibular ducts to the posterior tonsilar pillar. This procedure has been carried out on 53 patients over the past 15 years at the Childrens Hospital, Dublin. All patients have been followed up with a detailed questionnaire to determine symptomatic improvement, parent satisfaction and complications. Parental satisfaction regarding this procedure is high, with 94% of parents stating that their child had benefited from the operation and over half the parents reported complete cessation of all drooling within 3 months of the operation. The major complication of post-operative pneumonia presumed secondary to salivary aspiration occurred in three children. These patients all made a full recovery. Early minor complications occurred in two children, involving post operative submandibular gland swelling, and the late complication of a ranula was seen in four patients. We believe this is a safe and highly successful procedure that can significantly improve the quality of life of these children. PMID- 9222639 TI - Advanced glycation end products and their receptors co-localise in rat organs susceptible to diabetic microvascular injury. AB - Advanced glycation end products (AGEs) are believed to play an important role in the development of diabetic complications. AGEs are increased in experimental diabetes and treatment with the inhibitor of advanced glycation end products, aminoguanidine, has been shown to attenuate the level of these products in tissues undergoing complications. Recently, an AGE-binding protein has been isolated from bovine lung endothelial cells and termed the receptor for advanced glycated end products (RAGE). The present study sought to determine the distribution of AGE and RAGE in tissues susceptible to the long-term complications of diabetes including the kidney, eye, nerve, arteries as well as in a tissue resistant to such complications, the lung. Using polyclonal antisera both AGE and RAGE were found to co-localize in the renal glomerulus. AGE staining was clearly increased with age and was further increased by diabetes. Aminoguanidine treatment reduced AGE accumulation in the kidney. Co-localisation of AGE and RAGE was demonstrated in the inner plexiform layer and the inner limiting membrane of the retina and in nerve bundles from mesenteric arteries. In the aorta, both AGE and RAGE were found in the intima, media and adventitia. Medial staining was increased in diabetes and was reduced by aminoguanidine treatment. A similar pattern was observed for RAGE in the aorta. In the lung, RAGE was found widely distributed throughout the lung whereas the distribution of AGE staining was more limited, primarily localising to macrophages. The co localisation of AGEs and RAGE in sites of diabetic microvascular injury suggests that this ligand-receptor interaction may represent an important mechanism in the genesis of diabetic complications. PMID- 9222640 TI - Antioxidant treatment of experimental diabetic retinopathy in rats with nicanartine. AB - In order to study the contribution of oxidant stress to the pathogenesis of experimental diabetic retinopathy, male streptozotocin diabetic Lewis rats were treated with the antioxidant and lipid-lowering compound nicanartine (80 mg/kg; n = 8, blood glucose level 16.7 +/- 3.9 mmol/l; HbA1 11.8 +/- 1.6%) by oral supplementation for 6 months and compared with untreated diabetic (n = 6; blood glucose 18.1 +/- 1.4 mmol/l; HbA1 11.5 +/- 1.5%) and untreated, non-diabetic rats (n = 8; blood glucose 4.0 +/- 0.6 mmol/l; HbA1 4.8 +/- 0.9%). Diabetic retinopathy was evaluated by computer-assisted quantitative morphometry including measurement of autofluorescent advanced glycated end-products and immunohistochemistry for heme oxygenase I. Antioxidant treatment did not inhibit the 3.1-fold increase of retinal advanced glycation end products, while the expression of heme oxygenase I in both vascular and glial structures was markedly reduced. Chronic hyperglycaemia led to a 37.3% increase in endothelial cells (p < 0.001 vs normal controls) and a 26.6% reduction in pericyte numbers (p < 0.001 vs controls). Both abnormalities were significantly ameliorated by nicanartine (p < 0.001 vs diabetic controls). No effect was observed on the formation of acellular capillaries or microaneurysms. These data indicate that antioxidant therapy with nicanartine is of limited benefit in diabetic retinopathy, at least in the rodent model of streptozotocin-induced diabetes. PMID- 9222641 TI - Immunostimulation increases the resistance of mouse embryos to the teratogenic effect of diabetes mellitus. AB - The present work was aimed to assess the possible effect of stimulation of the maternal immune system on the teratogenic potential of diabetes mellitus. ICR female mice were immunized with splenocytes of male rats 3 weeks before the beginning of mating and were injected with 240 mg/kg streptozocin (STZ) 10 days after immunization. Females with blood glucose levels over 27.8 mmol/l and HbA1c levels over 6 standard deviations (SD) above the mean of intact animals were used for teratological studies. The rate of malformed fetuses, resorptions and fetal weights were evaluated for animals killed on day 19 of pregnancy using routine teratological methods. Also, phenotyping of spleen cells of these females was performed by fluorescein activated cell sorter analysis. Two main effects possibly due to immunostimulation of ICR females were observed: 1) immunostimulated females had significantly fewer litters with malformed fetuses than non-immunized females: only 4 litters out of 22 (18%) compared to 10 out of 16 (63%). Correspondingly, the incidence of malformed fetuses was also decreased: 2.1 compared to 8.9%; 2) a significant increase in the pregnancy rate in immunized diabetic ICR mice: 69% as compared to 44% in non-immunized diabetic females. Also, immunostimulation resulted in a visible increase in spleen cellularity and a certain increase in the number of cells with mature T-cell and macrophage surface markers. These results strongly suggest that immunostimulation increases the tolerance of ICR females to the teratogenic effect of STZ-induced diabetes. PMID- 9222642 TI - Suppression of cyclophosphamide induced diabetes development and pancreatic Th1 reactivity in NOD mice treated with the interleukin (IL)-12 antagonist IL 12(p40)2. AB - The macrophage product interleukin (IL)-12 is known to drive Th1 reactions in physiological and pathological immune responses. Here we report that treatment with the homodimeric IL-12p40 subunit, an antagonist of the bioactive IL 12p35/p40 heterodimer, suppresses diabetes development in cyclophosphamide injected NOD mice. Female mice of 70 days old received cyclophosphamide (250 mg/kg) to accelerate and synchronize diabetes development, and daily injections of 1 microgram IL-12(p40)2. While there was no delay of the first diabetes cases, the incidence of overt diabetes was significantly decreased in treated mice (46 vs 23%, p < 0.05). Analysis of mRNA expression in the pancreas showed that administration of the IL-12 antagonist had dampened interferon-gamma gene expression, decreased the ratio of interferon-gamma/IL-10 mRNA levels and in parallel suppressed the expression of the inducible nitric oxide synthase. At the same time intra-islet infiltration was significantly decreased (p < 0.001). Interestingly, the administration of IL-12(p40)2 also affected IL-12 gene expression, by downregulation of p35 mRNA. We conclude that IL-12 p40 homodimer suppresses diabetes development in the NOD mouse by dampening islet inflammation via selective down-regulation of Th1 type responses. The naturally occurring IL 12 antagonist IL-12(p40)2 represents a new and specific Th1 directed approach to prevent autoimmune diabetes. PMID- 9222643 TI - Relationships between plasma measures of oxidative stress and metabolic control in NIDDM. AB - Diabetes mellitus may be associated with increased lipid peroxidation which may contribute to long-term tissue damage. To test this hypothesis, we measured hydroperoxides (ROOHs) as well as alpha-tocopherol in plasma from healthy subjects and individuals with non-insulin-dependent diabetes mellitus (NIDDM) (n = 41 and 87, respectively). ROOHs were analysed using the ferrous oxidation with xylenol orange version II (FOX2) assay in conjunction with a specific ROOH reductant, triphenylphosphine, alpha-Tocopherol was analysed by HPLC with fluorimetric detection. NIDDM patients had lower cholesterol standardised alpha tocopherol levels as compared to control subjects (3.3 +/- 1.0 vs 5.1 +/- 2.3 (mumol/l)/(mmol/l); p < 0.0005, Mann-Whitney test): range (1.5-6.5 vs 1.9-13.0, respectively). Plasma ROOHs were substantially higher in the diabetic subjects compared to those of the control subjects (9.4 +/- 3.3 vs 4.1 +/- 2.2 mumol/l; p < 0.0005 Mann-Whitney test: range 2.7-16.8 vs 0.4-10.3, respectively). ROOH/cholesterol standardised alpha-tocopherol ratio was significantly higher in the diabetic patients compared to control subjects (3.2 +/- 1.6 vs 0.9 +/- 0.6; p < 0.0005, Mann-Whitney test: range 0.7-8.3 and 0.1-2.7, respectively). Plasma levels of ROOHs and alpha-tocopherol were similar in diabetic patients with or without complications as well as in smokers and non-smokers. The present study confirms previous findings from this laboratory that NIDDM is associated with increased oxidative stress as assessed by plasma ROOHs. Increased oxidative stress in diabetic patients appears to be related to the underlying metabolic abnormalities in diabetes, rather than to the complications of this disease. We therefore suggest that oxidative stress is an early stage in the disease pathology, which may contribute to the development of complications. PMID- 9222644 TI - Elevated sodium-lithium countertransport activity in erythrocytes is predictive of the development of microalbuminuria in IDDM. AB - Pathogenetic mechanisms other than the quality of metabolic control may play a role in the development of diabetic nephropathy. Some cross-sectional studies have shown that elevated erythrocyte sodium-lithium countertransport (Na+/Li+ CT) activity may be linked to incipient or overt nephropathy in insulin-dependent diabetic (IDDM) patients. The aim of the present work was to ascertain if high erythrocyte Na+/Li+ CT activity anticipates the development of microalbuminuria in IDDM patients. Evaluation of this cation transport system was carried out in 159 normotensive, normoalbuminuric IDDM patients, who were divided into two groups: those with values above (Group A) and those with values below (Group B) the median level in the overall population (300 mumol/erythrocytes x h). A total of 79 patients in Group A and 80 in Group B underwent periodic examinations over a similar time period (5.2 years, range 3.3-7.4 years and 5.4 years, range 3.4 7.5 years, respectively). Median sodium-lithium countertransport activity was stable when evaluated after 2 and 4 years of follow-up. Only seven patients were excluded from the protocol because changes in their sodium-lithium countertransport activity placed them on the other side of the median value with respect to their baseline measurement. Thus, 152 patients completed the study (76 in Group A and 76 in Group B). Of the 76 patients in Group A, 17 developed persistent microalbuminuria (22.3%). The number of patients in Group B showing persistent microalbuminuria was significantly lower (4 of 76; 5.2%; p < 0.01). The sensitivity of erythrocyte Na+/Li+ CT in predicting the development of microalbuminuria was 85% and its specificity was 55%. Seven patients of Group A and five of Group B developed arterial hypertension. Subjects in Group A had significantly higher mean HbA1c values of twice yearly measurements than those in Group B (9.6 +/- 1.7 vs 8.3 +/- 1.7%, p < 0.002, mean +/- SD) despite similar daily insulin requirements. Systolic and diastolic blood pressure levels were also evaluated every 6 months and were significantly higher in the Group A than in the Group B patients, although on average within the normal range. The odds ratio for developing persistent microalbuminuria in IDDM with elevated baseline erythrocyte Na+/Li+ CT activity after adjustment for gender and baseline albumin excretion rate, and mean 6 monthly plasma creatinine, HbA1c and systolic and diastolic blood pressure levels was 4.2 (95% confidence intervals 2.0-11.1). It was also found that the percentage of offspring having both parents with Na+/Li+ CT activity above the median value was significantly higher in Group A than in Group B (Group A vs Group B: 35 vs 19%; p < 0.01). On the contrary the percentage of offspring whose erythrocyte Na+/Li+ CT was lower in both parents was lower in Group A than in Group B: 10 vs 38%, p < 0.01). Parents of Group A offspring had arterial hypertension more frequently than those of Group B. These results indicate that erythrocyte Na+/Li+ CT activity is a useful diagnostic tool in identifying normotensive, normoalbuminuric patients who may be predisposed to develop persistent microalbuminuria. This disorder in the cation transport system is associated with poor metabolic control, higher blood pressure, and male sex; it also appears to be, at least partly, genetically transmitted. PMID- 9222646 TI - Genetic studies of neuropeptide Y and neuropeptide Y receptors Y1 and Y5 regions in morbid obesity. AB - Synthesis and release of neuropeptide Y (NPY) are both regulated by leptin binding to its hypothalamic receptor mediating some of the effects of leptin on food intake. Moreover, NPY administration is a powerful stimulant of feeding behaviour. Thus, we investigated the potential implication of NPY, NPY-Y1 and -Y5 subtype receptors [rNPY-Y1/-Y5] in the development of human obesity. Two complementary genetic approaches were used: 1) linkage analyses between obesity and polymorphic markers located nearby NPY and rNPY-Y1/-Y5 genes (respectively on chromosomes 7p15.1 and 4q[31.3-32]) in 93 French Caucasian morbidly obese families; 2) single strand conformation polymorphism (SSCP) scanning of the coding region of the NPY and rNPY-Y1 genes performed in 50 unrelated obese patients ascertained on the basis of a body mass index of 27 kg/m2 or more and a family history of obesity. No evidence of linkage between morbid obesity or obesity-related quantitative traits and NPY and rNPY-Y1/ Y5 regions was found in this population. Moreover, SSCP scanning revealed no mutation in the coding region of NPY and rNPY-Y1 genes among obese subjects. These results suggest that NPY and NPY-Y1/ Y5 receptors are unlikely to be implicated in the development of human morbid obesity, at least in the French Caucasian population. PMID- 9222645 TI - Oxidized lipoproteins found in patients with NIDDM stimulate radical-induced monocyte chemoattractant protein-1 mRNA expression in cultured human endothelial cells. AB - Although oxidized low density lipoprotein (LDL) exists in plasma from diabetic patients, there are few studies on its biological activity. Thus, we investigated the biological potency of LDL plus intermediate density lipoprotein fraction isolated from 12 non-diabetic and 24 non-insulin-dependent diabetic subjects of similar age and body mass index, in order to induce monocyte chemoattractant protein-1 (MCP-1) mRNA expression in cultured human endothelial cells. MCP-1 mRNA content in the cells exposed to the lipoproteins isolated from the diabetic patients was significantly higher than that from the control subjects (p < 0.001). The increment of MCP-1 mRNA content was positively correlated with not only HbA1c (r = 0.58, p < 0.0001) but also lysophosphatidylcholine (LPC) content in the lipoprotein (r = 0.46, p < 0.005) and was negatively correlated with diene formation lag time as a marker of oxidizability of the lipoprotein (r = -0.33, p < 0.05). Treatments of the cells with either 50 mumol/l probucol, 50 mumol/l alpha-tocopherol, or 0.1 mmol/l deferoxamine suppressed the increase in MCP-1 mRNA content induced by diabetic lipoproteins, respectively. Furthermore, the diabetic lipoproteins activated nuclear transcription factor NF-kappa B in the cells, which was inhibited by pre-treatment of cells with 50 mumol/l probucol. These data indicate that oxidatively modified lipoproteins found in diabetic plasma stimulate MCP-1 gene expression in endothelial cells. The LPC content which reflects oxidative modification of lipoprotein is at least a possible marker of biological activity to increase an atherogenic cytokine in endothelial cells. PMID- 9222647 TI - Renal function and insulin resistance as determinants of plasma leptin levels in patients with NIDDM. AB - Plasma leptin level is known to correlate with the degree of obesity. To determine the influences of renal function and insulin resistance on plasma leptin concentrations, we measured plasma leptin concentrations and performed the euglycaemic hyperinsulinaemic clamp studies in 57 patients with non-insulin dependent diabetes mellitus with a wide range of renal function. In simple regression analyses, plasma leptin concentration showed significant positive correlations with percentage of body fat measured by dual energy X-ray absorptiometry, body mass index, waist to hip ratio and fasting plasma insulin. Leptin level was higher in females than males. Multiple regression analyses indicated that percent body fat, waist to hip ratio, plasma insulin, gender and renal function (1/creatinine), but not insulin sensitivity, were significant and independent determinants of plasma leptin level. These results suggest that plasma leptin level is regulated or affected by multiple factors including renal function. Insulin resistance appeared to increase leptin levels indirectly by raising plasma insulin. PMID- 9222648 TI - Glucose tolerance and mortality, including a substudy of tolbutamide treatment. AB - Mortality according to glucose tolerance was studied to determine the prognosis of impaired glucose tolerance. Among 2500 persons tested in a community screening programme in 1962-1965 and followed-up for mortality to the end of 1987, age-sex adjusted mortality rates were 37.9 +/- 1.9, 53.6 +/- 4.2, and 70.1 +/- 3.6 deaths per 1000 person-years (+/-SE) in those with normal glucose tolerance, impaired glucose tolerance, and diabetes by World Health Organization criteria at baseline. Age-sex-adjusted mortality rates due to ischaemic heart disease were 14.3 +/- 1.1, 16.3 +/- 2.4, and 25.8 +/- 2.0 deaths per 1000 person-years, respectively. Using criteria predating those of the World Health Organization 147 men with abnormal glucose tolerance were entered into a randomized clinical trial in which 49 were treated with tolbutamide for approximately 10 years. Those treated had lower mortality rates from all causes (mortality rate ratio = 0.66, 95% confidence interval = 0.39, 1.10) and from ischaemic heart disease (mortality rate ratio = 0.42, 95% confidence interval = 0.16, 1.12) than those not receiving tolbutamide. Thus mortality rates are increased in persons with impaired glucose tolerance and diabetes, and the small clinical trial suggests that tolbutamide may be beneficial in men with abnormal glucose tolerance. PMID- 9222649 TI - Studies on the mass action effect of glucose in NIDDM and IDDM: evidence for glucose resistance. AB - The ability of hyperglycaemia to enhance glucose uptake was evaluated in 9 non insulin-dependent (NIDDM), 7 insulin-dependent (IDDM) diabetic subjects, and in 6 young and 9 older normal volunteers. Following overnight insulin-induced euglycaemia, a sequential three-step hyperglycaemic clamp (+ 2.8 + 5.6, and + 11.2 mmol/l above baseline) was performed with somatostatin plus replacing doses of basal insulin and glucagon, 3-3H-glucose infusion and indirect calorimetry. In the control subjects as a whole, glucose disposal increased at each hyperglycaemic step (13.1 +/- 0.6, 15.7 +/- 0.7, and 26.3 +/- 1.1 mumol/kg.min). In NIDDM (10.5 +/- 0.2, 12.1 +/- 1.0, and 17.5 +/- 1.1 mumol/kg.min), and IDDM (11.2 +/- 0.8, 12.9 +/- 1.0, and 15.6 +/- 1.1 mumol/kg.min) glucose disposal was lower during all three steps (p < 0.05-0.005). Hepatic glucose production declined proportionally to plasma glucose concentration to a similar extent in all four groups of patients. In control subjects, hyperglycaemia stimulated glucose oxidation (+4.4 +/- 0.7 mumol/kg.min) only at +11.2 mmol/l (p < 0.05), while non-oxidative glucose metabolism increased at each hyperglycaemic step (+3.1 +/- 0.7; +3.5 +/- 0.9, and +10.8 +/- 1.7 mumol/kg.min; all p < 0.05). In diabetic patients, no increment in glucose oxidation was elicited even at the highest hyperglycaemic plateau (IDDM = +0.5 +/- 1.5; NIDDM = +0.2 +/- 0.6 mumol/kg.min) and non-oxidative glucose metabolism was hampered (IDDM = +1.8 +/- 1.5, +3.1 +/- 1.7, and +4.3 +/- 1.8; NIDDM = +0.7 +/- 0.6, 2.1 +/- 0.9, and +7.0 +/- 0.8 mumol/kg.min; p < 0.05-0.005). Blood lactate concentration increased and plasma non-esterified fatty acid (NEFA) fell in control (p < 0.05) but not in diabetic subjects. The increments in blood lactate were correlated with the increase in non-oxidative glucose disposal and with the decrease in plasma NEFA. IN CONCLUSION: 1) the ability of hyperglycaemia to promote glucose disposal is impaired in NIDDM and IDDM; 2) stimulation of glucose oxidation and non-oxidative glucose metabolism accounts for glucose disposal; 3) both pathways of glucose metabolism are impaired in diabetic patients; 4) impaired ability of hyperglycaemia to suppress plasma NEFA is present in these patients. These results suggest that glucose resistance, that is the ability of glucose itself to promote glucose utilization, is impaired in both IDDM and NIDDM patients. PMID- 9222650 TI - Fibrinogen and von Willebrand factor in IDDM: relationships to lipid vascular risk factors, blood pressure, glycaemic control and urinary albumin excretion rate: the EURODIAB IDDM Complications Study. AB - The interrelationships between fibrinogen, von Willebrand factor, a marker of vascular endothelial cell damage, and serum lipids were explored in well characterised subjects with insulin-dependent diabetes mellitus. The 2091 subjects were enrolled into a cross-sectional, clinic-based study of complications, from 16 European countries: the EURODIAB IDDM Complications study. The anticipated significant relationships between both plasma fibrinogen and plasma von Willebrand factor concentrations and age and glycaemic control, and between fibrinogen and body mass index, were noted. Fibrinogen, adjusted for age and glycated haemoglobin concentration, was also related to smoking habits and was higher in the quartiles with highest systolic and diastolic blood pressures. There was a clustering of vascular risk factors, with a positive relationship between plasma fibrinogen and serum triglyceride concentrations in both genders and between fibrinogen and total cholesterol in males. An inverse relationship between fibrinogen and high density lipoprotein cholesterol was also apparent in males. A prominent feature was a positive relationship between both fibrinogen and von Willebrand factor and albumin excretion rate (p < 0.001 and p < 0.003 respectively) in those with retinopathy but not in those without this complication. In view of previous observations on blood pressure and albuminuria in these subjects the findings are consistent with the hypothesis that microalbuminuria and increased plasma von Willebrand factor are due to endothelial cell perturbation in response to mildly raised blood pressure in subjects with retinopathy. Fibrinogen may also contribute to microvascular disease and its relationships to lipid vascular risk factors suggest a possible pathogenic role in arterial disease in diabetes. PMID- 9222651 TI - Association between Ala54Thr substitution of the fatty acid-binding protein 2 gene with insulin resistance and intra-abdominal fat thickness in Japanese men. AB - Alanine to threonine substitution at codon 54 of the fatty acid-binding protein 2 (FABP2) gene was recently shown to be associated with insulin resistance in Pima Indians. It has been hypothesized that the mutation may result in enhanced intestinal up-take of fatty acids, and thereby an impairment of insulin action. We analysed the association of the Ala54Thr substitution with insulin sensitivity and abdominal fat thickness in 395 Japanese men aged 50.5 +/- 8.8 years (mean +/- SD) with a body mass index of 24.4 +/- 3.0 kg/m2. The frequency of the Thr54 allele was 0.34. Although the polymorphism was not significantly associated with diabetes or impaired glucose tolerance, subjects homozygous for the Thr54 allele had higher basal insulin levels. Analysis by homeostasis model assessment showed an association between the amino acid substitution and greater insulin resistance, and slightly higher beta-cell function. Oral glucose tolerance tests performed in 392 subjects without fasting hyperglycaemia showed higher 2-h insulin concentrations in individuals homozygous for the Thr54 allele when compared with heterozygotes or homozygotes for the Ala54 allele. No significant association was obtained between the polymorphism of the FABP2 gene and body mass index. However, ultrasound measurements of abdominal fat thickness revealed a greater accumulation of intra-abdominal fat in subjects homozygous for the Thr54 allele, whereas subcutaneous fat thickness was not associated with the polymorphism. These observations suggest that the Ala54Thr substitution in the FABP2 gene is associated with insulin resistance in Japanese men, and that visceral fat accumulation might be involved in the impaired insulin action associated with the substitution. PMID- 9222652 TI - Relatively more atherogenic coronary heart disease risk factors in prediabetic women than in prediabetic men. AB - Men with non-insulin-dependent diabetes mellitus (NIDDM) have a twofold increased risk of coronary heart disease and women with NIDDM have a fourfold increased risk. The reasons for this higher relative risk in NIDDM women than in NIDDM men is not completely understood. Since some studies suggest that duration of clinical diabetes and degree of hyperglycaemia have only a modest effect on coronary heart disease risk, we hypothesized that women who eventually convert to NIDDM might have a more atherogenic pattern of lipids and blood pressure relative to subjects who do not convert than male converters, even in the prediabetic period. We examined this issue in Mexican-American subjects in the 8-year follow up of the San Antonio Heart Study. Seventy-nine out of 801 men converted to NIDDM compared to 133 out of 1131 women. In both men and women, conversion to NIDDM was significantly associated with increased body mass index, fasting insulin and glucose, higher triglyceride and blood pressure and lower high density lipoprotein (HDL) cholesterol. The relative differences between converters and non-converters was significantly greater for women than for men; this interaction term for gender by conversion status was statistically significant for fasting insulin, triglyceride, HDL cholesterol and diastolic blood pressure. Thus, the higher relative risk for coronary heart disease in women with NIDDM relative to men with NIDDM may be partially due to their greater burden of cardiovascular risk factors even prior to the onset of diabetes. PMID- 9222653 TI - 24-h blood pressure and autonomic function is related to albumin excretion within the normoalbuminuric range in IDDM patients. AB - Significant changes in both blood pressure, autonomic function and kidney ultrastructure are observed in insulin-dependent diabetic (IDDM) patients with microalbuminuria. Intervention strategies are evaluated at even earlier stages of disease. Identification of patients at risk of developing microalbuminuria must be based on a thorough knowledge of the relations between key pathophysiological parameters in patients with normoalbuminuria. The aim of the present study was to characterize the interactions of urinary albumin excretion (UAE), 24-h ambulatory blood pressure (AMBP), and sympathovagal balance in a large group of normoalbuminuric IDDM patients. In 117 normoalbuminuric (UAE < 20 micrograms/min) patients we performed 24-h AMBP (Spacelabs 90207), with assessment of diurnal blood pressure and heart rate (HR) variation, and short-term (three times 5 min) power spectral analysis of RR interval oscillations, as well as cardiovascular reflex tests (HR variation to deep breathing, postural HR and blood pressure response). Patients with UAE above the median (4.2 micrograms/min) had significantly higher 24-h systolic and diastolic AMBP (125 +/- 10.1/76 +/- 7.2 mmHg) compared to the low normolbuminuric group (120 +/- 8.4/74 +/- 5.1 mmHg), p < 0.01 and 0.02, respectively. Patients with UAE above the median had significantly reduced short-term RR interval variability including both the high frequency component (5.47 +/- 1.36 vs 6.10 +/- 1.43 ln ms2), and low frequency component (5.48 +/- 1.18 ln ms2 compared to 5.80 +/- 1.41 ln ms2), p < 0.02 and p = 0.04 (ANOVA). In addition, patients with high-normal UAE had reduced mean RR level (faster heart rates) 916 +/- 108 compared to 963 +/- 140 ms, p < 0.04. These differences were not explained by age, duration of diabetes, gender, level of physical activity, or cigarette smoking. HbA1c was significantly higher (8.6 +/- 1.2 vs 8.2 +/- 1.0%, p = 0.03) in the group with high normal UAE. Comparing normoalbuminuric IDDM patients with UAE above and below the median value, we found significantly higher AMBP in combination with significant differences in sympathovagal balance and significantly poorer glycaemic control in the group with high-normal albumin excretion. Our data demonstrate interactions between albumin excretion, blood pressure, autonomic function, and glycaemic status, already present in the normoalbuminuric range and may describe a syndrome indicative of later complications. PMID- 9222654 TI - Ocular vascular endothelial growth factor levels in diabetic rats are elevated before observable retinal proliferative changes. AB - Vascular endothelial growth factor (VEGF) is a potent angiogenic factor. VEGF levels in ocular tissue of 6-, 12-, 18- and 28-week-old Goto-Kakizaki (GK) rats, a well-known model of non-insulin-dependent diabetes, were evaluated by highly sensitive ELISA. VEGF concentrations in the GK rat as well as in non-diabetic Wistar rat significantly decreased from the age of 6 weeks to 18 weeks. However, although VEGF concentrations in the Wistar rat continued to fall significantly from 18 to 28 weeks of age, the levels were maintained between 18 and 28 weeks of age in GK rats. Levels were significantly different between the GK and Wistar rats at 28 weeks of age. Results of immunohistochemical studies of the eyes of Wistar and GK rats at 28 weeks of age suggest diffuse distribution of this cytokine in cells of neural origin. Weak to moderate VEGF immunoreactivity was exhibited mainly in the ganglion cell layer, inner plexiform layer and inner/outer nuclear layers in rats with and without diabetes. However, in the retinal optic nerve fiber layer, retinal pigment epithelium and choroid, strong VEGF immunoreactivity was noted only in the GK rat. In conclusion, increased VEGF production in certain ocular tissue, similar to that in humans, is observed quite early, at least before the appearance of observable retinal changes in the diabetic GK rat. This also suggests that the GK rat can be used as a model of initial or latent phase diabetic retinopathy. PMID- 9222655 TI - Fibrinogen and diabetes mellitus: is it time for intervention trials? PMID- 9222656 TI - Fibrin(ogen) and diabetes mellitus: don't forget fibrinolysis. PMID- 9222657 TI - Fibrinogen, fibrinolysis and diabetes mellitus: a comment. PMID- 9222658 TI - Thiamine may indirectly act as an antioxidant. PMID- 9222660 TI - Treatment of esophageal varices. PMID- 9222659 TI - Prevention of IDDM: strategies based on new observations of molecular pathogenesis. Workshop held in Majvik, Kirkkonummi, Finland, 6-8 May 1996. PMID- 9222661 TI - Recent situation in drug treatment for esophageal varices. PMID- 9222662 TI - Elastic band ligation for bleeding esophagogastric varices. AB - Experimental studies have shown ligation results in replacement of submucosal structures, including varices, with scar tissue resulting in eradication of varices. Elastic band ligation is equal in efficacy to sclerotherapy for control of active bleeding from esophageal varices. Ligation appears superior to sclerotherapy for long-term prevention of recurrent variceal bleeding, requires fewer treatment sessions to eradicate varices, is associated with fewer bleeding and non-bleeding complications of treatment, and results in improved survival. Results from the synchronous combination of elastic band ligation with sclerotherapy do not appear to be superior to those obtainable with those from endoscopic ligation used alone. PMID- 9222663 TI - Prophylactic endoscopic sclerotherapy in patients with liver cirrhosis, portal hypertension, and esophageal varices. AB - A major cause of death among patients with cirrhosis and portal hypertension is bleeding from esophago varices. The reasons for variceal haemorrhage and which varices have a high risk of bleeding are discussed. In addition, the question as to whether prophylactic endoscopic sclerotherapy is a viable solution to prevent variceal haemorrhage. PMID- 9222664 TI - Endoscopic treatment for esophago-gastric varices, current status in Japan. AB - BACKGROUND/AIMS: Endoscopic treatment for esophago-gastric varices is performed to save patients who suffer from bleeding. The purpose of this paper is to introduce the recent status in Japan and our original therapeutic measures and results. MATERIALS AND METHODS: Japan's current status is discussed by our society's surveillant study and references. Our techniques' and results are also described through 214 patients which were treated by combination of endoscopic variceal ligation and sclerotherapy as well as 65 patients who underwent endoscopic injection sclerotherapy using histoacryl. Combination therapy is performed by endoscopic ligation and sclerotherapy using polidocanol and Histoacryl, without any contrast agent. RESULTS: Combination therapy shaved a 97% eradicating effect which was as high as sclerotherapy alone (92%) and which required less therapeutic sessions than sclerotherapy. For actively bleeding patients, endoscopic ligation showed 100% hemostatic rate (29/29) while conventional sclerotherapy obtained 92% hemostasis (86/93). Histoacryl injection that were employed for uncontrollable bleeding from gastric varices and early recurrent bleeding of esophageal varices obtained 100% hemostatic rate. CONCLUSIONS: Our original combination therapy was superior to either endoscopic ligation and sclerotherapy alone and histoacryl injection showed better results in hemostatic effect than conventional sclerotherapy without any complications. PMID- 9222665 TI - Recent position of transjugular intrahepatic portosystemic shunt in the treatment of portal hypertension. AB - Transjugular intrahepatic portosystemic shunt (TIPS) is a side-to-side portocaval shunt for threatening complications of portal hypertension. TIPS effectively decreases portal hypertension connecting the hepatic and portal vein with an expandable metal stent without the mortality and morbidity of an open surgical procedure. Technical success can be achieved in over 90% of patients, with procedure related mortality of 1-2%. The main problem is stenosis or occlusion of the shunt by neo-intimal hyperplasia narrowing the lumen of the shunt in 20-80% of patients during 6-12 months, but fortunately, most stenotic stents can be revised successfully. Recent indications for TIPS are acute variceal hemorrhage refractory to endoscopic treatment and recurrent variceal bleeding despite sclerotherapy or band ligation. TIPS insertion in the treatment of refractory ascites seems to be promising. PMID- 9222666 TI - Effects of octreotide and a-tocopherol on bacterial translocation in experimental intestinal obstruction: a microbiological, light and electronmicroscopical study. AB - BACKGROUND/AIMS: Bacterial translocation induced by intestinal obstruction is suggested to be due to increased intestinal luminal volume, leading to intestinal overgrowth with certain enteric microorganisms and intestinal mucosal damage. If this suggestion is true, maintenance of intestinal mucosal integrity by a cytoprotective agent, a-tocopherol, and inhibition of gastrointestinal secretions by octreotide should decrease the incidence of bacterial translocation and extent of mucosal injury due to intestinal obstruction. METHODS: Complete intestinal obstruction was created in the distal ileum of male Wistar Albino rats by a single 3-0 silk suture. The animals received subcutaneous injections of 1 ml of physiologic saline (group 1) (PS 24) and 1 ml of saline containing octreotide acetate (100 micrograms/kg) (group 2) (OC 24), at 0, 12 and 24 hours of obstruction. In group 3 (PS 48) and group 4 (OC 48), the rats were treated with subcutaneous physiologic saline (1 ml) and octreotide acetate (100 micrograms/kg), respectively, beginning at the time of obstruction and every 12 hours for 48 hours. The rats in group 5 (Toc 24), were pretreated with intramuscular a-tocopherol 500 mg/kg on day 1 and 8, and underwent laparotomy on day 9. A third dose of a-tocopherol was injected at the time of obstruction on day 9 and no treatment was given thereafter. We tested the incidence of bacterial translocation in systemic organs and circulation and evaluated the histopathological changes in all groups. RESULTS: Treatment with octreotide acetate was found to be ineffective in reducing the incidence of translocation, with no histopathological improvement. Mucosal damage scores, on the other hand, in the a-tocopherol group were statistically less than those in the octreotide and control groups (p < 0.05). Additionally, a-tocopherol treatment decreased the incidence of organ invasion with translocating bacteria, although this difference did not reach statistical significance. CONCLUSION: Octreotide acetate treatment in complete intestinal obstruction has no effect on the incidence of bacterial translocation. a-Tocopherol, on the other hand, has a cytoprotective effect on intestinal mucosa in intestinal obstruction which, in turn, is thought to decrease bacterial translocation when used in physiological doses and prophylactically. PMID- 9222667 TI - Centrocytic lymphoma presenting as a subphrenic abscess and a solitary liver nodule. AB - Non Hodgkin's lymphoma revealed by hepatic manifestations is extremely rare. We describe here a 82-year old male patient who presented with a right subphrenic abscess and a solitary liver tumour that was shown to be a centrocytic lymphoma. Furthermore, asymptomatic cryptogenic liver cirrhosis was diagnosed. This previously unreported form of clinical presentation of a non Hodgkin's lymphoma as well as the association with liver cirrhosis are discussed in the context of the recent literature. PMID- 9222668 TI - Extrahepatic portal venous laceration in TIPS treated with stent graft placement. AB - Three cases of intraperitoneal bleeding from extrahepatic portal vein laceration were observed in a series of 104 patients with TIPS. In one patient, bleeding continued after Wallstent placement and the patient died during an emergency laparotomy. Bleeding was stopped in the other two patients by using a stent graft rather than a regular stent. One shunt remained patent and outflow stenosis developed in 3 months. The other shunt thrombosed in 5 months. Both shunts were fully reopened. PMID- 9222669 TI - Surgical risks of acute cholecystitis in elderly. AB - BACKGROUND/AIMS: For the elderly patient, an emergency biliary procedure carries a higher risk than an elective operation. Recently introduced advances in ultrasonography and critical care medicine have affected the clinical risks of surgery for acute cholecystitis in the elderly. This study evaluated the clinical risks of open cholecystectomy for the elderly with acute cholecystitis. MATERIALS AND METHODS: During a 10 year period (1985-1994), a total of 145 patients were diagnosed with acute cholecystitis and underwent cholecystectomy. According to their age, the patients were divided into 3 groups (Group A < 59 years of age; Group B between 60-69 years of age; Group C > 70 years of age). The characteristics and the surgical risk factors in open cholecystectomy for the elderly with acute cholecystitis were evaluated. RESULTS: The rate of acalculous cholecystitis and choledochal stones were significantly (p < 0.05) high in Group C. Septic complication, gangrenous changes, and positive bile culture were also increased parallel to the increase in age. A noteworthy finding was an incidental carcinoma found in a case in group B and in 3 cases in group C. Hospital stay was significantly longer in Group C than in the other groups due to pre-operative complications and post-operative morbidity. CONCLUSION: With respect to increase in elderly patients with acute cholecystitis who present more frequent gangrenous changes and carcinomatous changes as well as high rate of septic complication, successful treatment of these patients is increased when early surgery can be performed on the basis of accurate and prompt diagnosis using imaging modalities and meticulous peri-operative management. PMID- 9222670 TI - Prolonged cholestatic jaundice after endoscopic retrograde cholangiography. AB - The main complications of endoscopic retrograde cholangiography and sphincterotomy are bleeding, pancreatitis, perforation and sepsis. Two cases of unexplained prolonged cholestatic jaundice in patients who underwent endoscopic retrograde cholangiography (ERC) for biliary obstruction due to choledocholithiasis are reported. The patients were admitted because of right upper quadrant pain, vomiting and jaundice. Laboratory tests showed increased levels of total and conjugated serum bilirubin and increased alkaline phosphatase. Ultrasound examination showed cholelithiasis and choledocholithiasis with bile duct dilatation. ERC with sphincterotomy was performed and gallstones obstructing the common bile duct were removed endoscopically. Following ERC and despite complete patency of the biliary tree, a progressive increase of total and conjugated bilirubin and of alkaline phosphatase was noted, associated with itching and total stool discoloration. The insertion of nasobiliary drain did not improve the jaundice. Prednisolone treatment for 12 days was associated with progressive restoration of serum bilirubin alkaline phosphatase to normal levels. It was postulated that the radiocontrast material used may have acted toxically on the liver with disruption of the canalicular plasma membrane. It is proposed that intrahepatic cholestasis should be added in the list of complications of endoscopic retrograde cholangiography. PMID- 9222671 TI - Villous adenoma of the gallbladder: a case report. AB - A 76-year-old female with villous adenoma of the gallbladder is herein reported. She presented complaining of upper abdominal pain for some months prior to admission. No tumor mass could be detected by transabdominal ultrasonography, but CT scan demonstrated a sessile tumor measuring some 2 cm in diameter. A simple cholecystectomy was thus performed. The resected gallbladder was filled with mucinous material, and showed a sessile tumor with finger-like processes in the fundus. Both the macro- and microscopical findings of the lesion were almost identical to a typical villous adenoma of the colon. Little information is available on villous adenoma of the gallbladder, because such cases are extremely rare. In this paper, is described e a rare villous adenoma lesion while paying special attention to the morphological findings. PMID- 9222672 TI - Erythropoietic response induced by recombinant human erythropoietin in anemic cancer patients candidate to major abdominal surgery. AB - BACKGROUND/AIMS: This trial was designed to quantify the production of new red blood cells (RBCs) after treatment with recombinant human erythropoietin (r HuEPO). MATERIALS AND METHOD: Twenty anemic patients with gastric or colorectal cancer were prospectively studied. The control group (n = 10) underwent surgery without delay. The experimental group (n = 10) received treatment with 500 U/kg of r-HuEPO divided in three doses which were administrated as follows: 300 U/kg 12 days before surgery and 100 U/kg 4 and 8 days later. RESULTS: RBC gain was 210 +/- 75 mL in the experimental group which received treatment. The erythropoietic response was similar after 300 U/kg (59 +/- 36 mL) or 100 U/kg 75 +/- 24 mL). No correlation was found between RBC gain and age (r = 0.18) or baseline levels of erythropoietin (r = 0.08), hemoglobin (r = 0.06), erythrocytes (r = 0.01), reticulocytes (r = 0.01), iron (r = 0.28) and ferritin (r = 0.08). One treated patient in the experimental group was transfused with allogeneic blood (2 units) versus 5 control patients (overall 13 units) (p = 0.06). CONCLUSION: Preoperative treatment with r-HuEPO improved erythropoiesis and reduced allogeneic transfusions in anemic cancer patients. No significant difference in RBC gains was noted between r-HuEPO administration at 300 U/kg and r-HuEPO administration at 100 U/kg. This result suggests a possible dose reduction for future trials. PMID- 9222673 TI - Restorative proctocolectomy: histological assessment and cytometric DNA analysis of ileal pouch biopsies. AB - BACKGROUND/AIMS: The pathological changes and the risk of developing cancer in the ileal pouch mucosa of patients who received restorative proctocolectomy with ileal pouch-anal anastomosis (IPAA) were studied. The presence or absence of remaining rectal mucosa below the IPAA in both patients with stapled and handsewn IPAA was also examined. MATERIALS AND METHODS: Endoscopy of the ileal pouch was performed on 38 patients at 4, 12, 18 and 36 months after restorative proctocolectomy with ileal pouch. Mucosal biopsy specimens were taken from the ileal reservoir in order to assess the histological incidence of inflammation. In 23 patients, biopsies were taken to perform cytometric DNA analysis. Clinical symptoms of pouchitis (over six evacuations in 24 hours, night-time evacuations, leakage of feces, bloody diarrhea, abdominal pain and fever) were recorded and correlated with the histological findings. Biopsies were also sampled below the ileo-anal anastomosis (IPAA) in order to identify residual rectal mucosa. RESULTS: Results of histological assessment showed various degrees of chronic inflammation increasing over time (from 42 to 60%) while the presence of both acute and chronic inflammation of the reservoir was less frequent (from 18 to 30%). Villous atrophy was present in 39-68% of patients and the grade of villous atrophy was correlated to the grade of inflammation. Clinical pouchitis was present in 3 to 8% of cases at the different controls and it was always associated with the highest grade of histological inflammation and severe villous atrophy. No significant alteration of the DNA cellular content was observed. Very low incidence of aneuploidy (0.7-1% Ex.R.) has been reported in three cases. However, we found dysplasia in only one patient who underwent surgical treatment for familial polyposis coli. IPAA evaluation showed no residual rectal mucosa in 40% of cases with stapled IPAA; in the remaining 60%, we found a small amount of rectal mucosa (maximum 1 cm). We did not find rectal mucosa after handsewn IPAA with mucosectomy. CONCLUSIONS: Patients treated with restorative proctocolectomy with IPAA showed a higher and increased incidence of inflammation during follow up. No significant alteration of DNA cellular content nor dysplasia of the pouch mucosa were observed. In this study the chance of leaving rectal mucosa after stapled IPAA was about 60%. PMID- 9222674 TI - Endoscopic resection of large sessile colorectal polyps using a submucosal saline injection technique. AB - BACKGROUND/AIMS: Endoscopic removal of sessile colorectal polyps 2 cm or greater in diameter is very difficult. We evaluated the safety and usefulness of submucosal saline injection in endoscopic resection of these polyps. MATERIALS AND METHODS: Under colonoscopic observation, 0.9% NaCl was injected submucosally through needle forceps to elevate polyps, which were then resected by electrocoagulation. Before 1989, the depth and healing status of ulcers produced by endoscopic resection with (N = 12) and without (N = 16) submucosal saline injection were examined in specimens obtained at subsequent colectomies. Between 1990 and 1993, patients were assigned by weighted randomization to undergo colonoscopic polypectomy with (N = 24) or without (N = 5) submucosal saline injection. RESULTS: All 12 ulcers after submucosal saline injection were confined to the submucosal layer while 7 (44%) of 16 ulcers without submucosal saline injection reached the muscle layer or deeper. Seventeen of 24 (71%) polyps were completely removed (14 piecemeal and 3 en bloc) without serious complications after saline injection. Resection without submucosal saline injection was complete, but piecemeal, for two of five polyps; postresection bleeding occurred in one case. CONCLUSIONS: Endoscopic resection after submucosal saline injection is a safe and effective treatment for large, sessile colorectal polyps. PMID- 9222675 TI - Normal rectal mucosa. Should we biopsy? AB - BACKGROUND/AIMS: To determine the practice of routine rectal biopsy in the United Kingdom, and assess the diagnostic yield and complications of rectal biopsy in patients presenting with diarrhea. MATERIALS AND METHODS: A postal questionnaire was sent to consultant members of the British Society of Gastroenterology. An audit of the diagnostic yield and complications resulting from routine biopsy of normal looking rectal mucosa in patients presenting with diarrhea was performed. RESULTS: Ninety five (35%) consultants "nearly always" biopsy normal looking mucosa, with a further 56 (20%) taking a biopsy in more than fifty percent of cases. Fifty five (20%) almost never biopsy if the mucosa looks normal, with 68 (20%) taking a biopsy less than fifty percent of the time. Biopsies were taken from 50 patients referred with diarrhea whose rectal mucosa looked normal. Abnormal histology was reported in 11 (22%) cases. The rectal biopsy led to a positive diagnosis and change in management in 4 (8%) cases. The remaining biopsies showed minor inflammatory changes that were not considered clinically important. One significant complication occurred due to rectal biopsy. CONCLUSIONS: Clinicians disagree on the value of routine rectal biopsy in the investigation of diarrhea. In patients presenting with diarrhea, the diagnostic yield from biopsy of normal looking rectal mucosa is low. Life threatening complications can occur and in unselected patients routine biopsy should not be performed. PMID- 9222676 TI - Endometriosis of the rectum causing bowel obstruction: a case report. AB - A 42-year-old pre-menopausal woman complaining of lower abdominal pain was referred to our hospital. A barium enema showed rectal stenosis and colonoscopy revealed that the mucosa at the stenotic site was normal with no cancerous changes. Pelvic computed tomography demonstrated an adhesion between the rectum and uterus and a thickened rectal wall. The patient underwent exploratory laparotomy under a diagnosis of rectal stenosis. The rectum was found to be surrounded by inflammatory fibrous tissue, which caused the stenosis. As no dissection plane was discernible between the rectum and uterus, low anterior resection of the rectum and hysterectomy were performed. Histological examination showed that endometrial-type glands extended circumferentially around the rectum and invaded the rectal submucosal layer and subsequently, endometriosis of the rectum was diagnosed. PMID- 9222677 TI - Colitis associated to chemotherapy with 5-fluorouracil. AB - Antitumoral agents, especially when administered in combination, can induce pseudomembranous colitis, due to their antimitotic and antibacterial properties. Although patients given 5-fluorouracil frequently show nonspecific colitis or diarrhea without colitis, very few cases of proven pseudomembranous colitis have been described during 5-fluorouracil monotherapy. We describe the second case reported in the English literature of a typical pseudomembranous colitis occurring in a patient given 5-fluorouracil as a single antimitotic agent for colonic cancer. Intestinal injury was not preceded by antibiotic therapy. Although both stool and pseudomembrane culture did not yield C. difficile, oral vancomycin was started. This treatment led to a prompt improvement of intestinal symptoms and colitis was resolved in one week. PMID- 9222678 TI - Intraluminal duodenal diverticulum with malposition of the ampulla of Vater. AB - Intraluminal duodenal diverticulum is a rare congenital anomaly, sometimes associated with malposition of the ampulla of Vater. When the diverticulum is excised, the position of the ampulla should be determined carefully to avoid injury to pancreaticobiliary ducts. We report two patients with symptomatic intraluminal duodenal diverticulum and malposition of the ampulla. The ampulla was located on the rim of the diverticulum in one patient; in the others, the ampullary site was the posterior wall of the duodenum. Both patients underwent successful excision of the diverticulum without ductal injuries. As we have been unable to find any case with an ampullary location on the anterior wall of the duodenum, anterior duodenotomy followed by identification of the ampulla must precede excision of the diverticulum in order to avoid pancreaticobiliary ductal injuries. PMID- 9222679 TI - Transthoraco-phrenic esophageal transection with paraesophago-gastric devascularization and splenectomy using a stapler. AB - BACKGROUND/AIMS: Surgery remains the most reliable treatment for bleeding esophageal varices. The aim of this study was to introduce the operative technique of transthoraco-phrenic esophageal transection with paraesophagogastric devascularization using a stapler and to evaluate surgical results. METHODS: Forty-five patients underwent the procedure; an elective procedure was performed in 22 patients (bleeders) and a prophylactic procedure in 23 patients (nonbleeders). Twenty-nine patients were classified as Child's A, 15 as B and 1 as C. Previous sclerotherapy had been performed in 5 patients. RESULTS: No hospital deaths occurred. No patients developed postoperative anastomotic leakage, encephalopathy, or any complications related to phrenicotomy. Three patients bled postoperatively from recurrent esophageal varices. Cumulative 5 year bleeding rates were 5.0% in bleeders and 6.6% in non-bleeders. Two patients died due to bleeding varices. Cumulative 5-year survival rates were 72.1% and 78.8% in patients classified as Child's A and Child's B, respectively. CONCLUSIONS: This procedure may be indicated for a majority of Child's A or B patients. Although the advantages of this procedure must be evaluated further, it may be an alternative when injection sclerotherapy and endoscopic ligation fail. PMID- 9222680 TI - Ultrasonographic imaging of neoplasms of the cervical esophagus. AB - BACKGROUND/AIMS: We studied the effectiveness of ultrasonography in evaluating the cervical esophagus for the presence of large masses arising from the esophageal wall and consequently, the modifications of the visceral lumen. MATERIALS AND METHODS: The cervical esophagus can be evaluated by ultrasound with longitudinal and axial scans, using the left thyroid lobe as an acoustic window. The cervical esophagus can be visualized from the C5 to D2 vertebrae. From November 1992 to July 1996, 220 patients with esophageal cancer and 120 subjects without esophageal disease (control group) were examined with ultrasonography. Examination of the cervical esophagus was performed with a linear high definition small parts probe with a frequency of 7.5-10 Mhz. RESULTS: In all 31 patients with cancer of the cervical esophagus, ultrasonography of the cervical region showed the presence of an expanding mass from the esophageal wall as well as the modifications in the visceral lumen. The neoplasm of the cervical esophagus was visualized when its diameter exceeded 5 mm. CONCLUSIONS: The experience of the authors shows that, during ultrasound examination of the cervical region, it is possible to accurately evaluate the cervical esophagus, either morphologically or functionally. PMID- 9222681 TI - Evaluation of leiomyoma of the esophagus with endoscopic ultrasonography. AB - Fourteen patients with suspected leiomyoma of the esophagus were studied by endoscopic ultrasonography, computed tomography, endoscopy and barium swallow. The results were correlated with the histology of the resected specimens: in 2 patients with a peduncolated leiomyoma originating from the second echographic layer, endoscopic resection was performed. Endoscopic ultrasonography was superior to other imaging techniques in detection and staging of leiomyoma because it can determine the layer of origin, the direction of the growth and the consistency of the tumor. PMID- 9222682 TI - Primary biliary cirrhosis: Dutch application of the Mayo Model before and after orthotopic liver transplantation. AB - BACKGROUND/AIMS: A retrospective study of primary biliary cirrhosis (PBC) was performed to study the Original Mayo Model for predicting survival by a Dutch data-set of patients, presentation of disease progression; assessment of liver transplantation, prediction of post-transplantation survival; and the addition of two laboratory variables to the Original Mayo Model. MATERIALS AND METHODS: Survival of 83 patients, 37 of whom underwent transplantation, were studied. Mean follow-up was 6.0 +/- 0.45 SEM years. Risk score at diagnosis, platelet count, and serum sodium were analyzed in a Cox model. RESULTS: The Original Mayo Model estimated survival for low-, medium-, and high-risk groups accurately and it also presented disease progression. Baseline Mayo risk score in a Cox model had a regression coefficient of 1.01, indicating an excellent predictor p < 0.0001. Platelet count was a predictor of survival (p < 0.002), whereas serum sodium did not (p = 0.67). A new model combined of the Original Mayo risk score and platelet count predicted survival in high-risk patients somewhat better compared to the Original Mayo Model. With both models, liver transplantation had a significant beneficial effect on survival (p < 0.001). The scores revealed no significant influence (p = 0.47) for overall post-transplantation survival. CONCLUSIONS: The Original Mayo Model remains the model of choice for patients with PBC for prognostication from 3-8 years, is a useful tool in the assessment of liver transplantation but not an indicator of post-transplantation survival. Platelet count showed to have additional prognostic value. A new model combined of platelet count and the Original Mayo risk score did predict survival in high-risk groups slightly better compared to the Original Mayo Model. PMID- 9222683 TI - Human erythrocyte polyamine levels after partial hepatectomy. AB - BACKGROUND/AIM: There are various indices of liver regeneration, but no clinically useful index that reflects the current status of liver regeneration. We assayed human erythrocyte polyamine levels after partial hepatectomy to define the relationship between erythrocyte polyamine levels and liver regeneration. MATERIALS AND METHODS: Levels of human erythrocyte polyamines (putrescine, spermidine, and spermine) were assayed by high-pressure liquid chromatography in 91 patients after partial hepatectomy and in 13 patients after surgery other than partial hepatectomy (controls). Of the patients after partial hepatectomy, 37 underwent hepatectomy of 20% or more of the liver (group A), 27 underwent segmentectomy or subsegmentectomy of the liver amounting to less than 20% of the liver (group B), and 27 underwent an operation smaller in scale than sub segmentectomy (group C). RESULTS: The greater the proportion of the liver resected, the greater was the percent increase. In groups A, B, and C, erythrocyte levels of spermidine and spermine increased after surgery compared with the base line, and were significantly higher at 7 or 14 days, decreasing later. The differences in spermidine among the three groups were significant. CONCLUSIONS: After partial hepatectomy, the erythrocyte polyamine levels, especially the level of spermidine, were related to the proportion of liver resected. They seemed to reflect the degree of liver regeneration. PMID- 9222684 TI - Resection of hepatic metastases from colorectal cancer. AB - BACKGROUND/AIMS: Long term results of hepatic resection for metastases from colorectal cancer depend upon several factors which are related to both features of primary cancer and of metastases. The aim of this study was to evaluate prognostic factors that best correlate with long-term results. MATERIALS AND METHODS: Fifty-eight hepatic resections were performed for colorectal cancer metastases. Long-term results were evaluated in relation to age of patients, features of primary tumor, features of metastases, section margin, number of intra-operative blood transfusions and execution of adjuvant chemotherapy. RESULTS: Overall 5-year survival rate was 17%. 5-year survival rate in patients with stage B primary tumor was 63%, in patients with late metachronous metastases it was 28%, in patients with section margin > 1 cm it was 33% and in patients who did not receive intra-operative transfusions it was 45%. Patients with a solitary metastasis or with metastases sized less than 4 cm and those who received adjuvant chemotherapy also showed a better survival than the others. CONCLUSIONS: Better results were observed in patients without nodal involvement of the primary tumor. Patients with a small solitary metachronous metastasis that appeared more than one year after the colorectal resection and resected with a section margin of more than 1 cm, also showed good results. PMID- 9222685 TI - Surgical treatment of cholangiocarcinoma. AB - BACKGROUND/AIMS: To report the results of surgical treatment of intrahepatic cholangiocarcinoma with different procedures and to find the factors that may affect the long-term survival. MATERIALS AND METHODS: From 1987 to 1994, 57 patients with intrahepatic cholangiocarcinoma underwent laparotomy. Among them, resection was performed in 27 patients, operative drainage in 14 patients and biopsy only in 14 patients. The liver resections included 9 right lobectomies, 14 left lobectomies and 4 hilar resections. All specimens were stained with carcinoembryonic antigen (CEA) and HLA-DR monoclonal antibodies. RESULTS: There were 7 postoperative mortalities, one in the resection group (1/27), two in the drainage group (2/14) and 4 in the biopsy group (4/14). Patients undergoing resection survived significantly longer (median, 8 months) (mean, 19 +/- 4 months) than patients undergoing drainage (median, 4 months) (mean, 6 +/- 2 months) and biopsy (median, 2 months) (mean, 3 +/- 1 months) (p < 0.01). After resection, univariate analysis showed that positive hiliar lymphnode was a poor prognostic sign and mucobilia was a good prognostic sign. Age, sex, size of tumors cell differentiation, clear margin, and positive HLA-DR and CEA had no effect on prognosis. CONCLUSION: The results support the surgical resection of intrahepatic cholangiocarcinoma. Tumor free margin should be aggressively achieved but may not be necessary. Mucobilia is a good prognostic sign and positive hilar lymphnode is a grave sign. PMID- 9222687 TI - Percutaneous transhepatic transluminal forceps biopsy in obstructive jaundice. AB - BACKGROUND/AIMS: To evaluate the technical feasibility and sensitivity of percutaneous transluminal forceps biopsy of bile duct diseases. MATERIAL AND METHODS: Seventeen fluoroscopic-guided transluminal forceps biopsies were performed in 16 patients with obstructive jaundice. The technique was performed through an existing percutaneous transhepatic tract. Multiple specimens were obtained after passing the forceps biopsy into a long 9-French sheath and the specimens were fixed with formalin for histopathologic diagnosis. RESULTS: Adequate samples for histological diagnosis was obtained in 12 of 17 procedures (sensitivity, 71%). Pathologic reports included pancreatic head carcinoma n = 2, cholangiocarcinoma n = 3, hepatoma with intrahepatic-bile duct invasion n = 3, common bile duct tumors n = 3 and chronic inflammation n = 1. Minor complications such as pain was noted in three patients while transient hemobilia was seen in two patients. CONCLUSIONS: Percutaneous transhepatic transluminal forceps biopsy is a safe technique which is easy to perform. This can be done through an existing transhepatic biliary tract with a sensitivity rate of 71%. PMID- 9222686 TI - Resistance to 3-mercaptopropionic acid-induced seizures in hepatic encephalopathy. AB - BACKGROUND/AIMS: To determine if a model of hepatic encephalopathy (HE) exhibits decreased sensitivity to the neuronal effects of a drug that induces seizures as a consequence of decreasing GABA-mediated inhibitory neurotransmission. MATERIALS AND METHODS: 3-Mercaptopropionic Acid (MPA) is an inhibitor of L-glutamate decarboxylase which catalyzes the synthesis of GABA from glutamate. MPA was administered, either by intraperitoneal or intracerebroventricular injection, into rats with stage III HE due to thioacetamide-induced fulminant hepatic failure and into normal control rats. RESULTS: When MPA was administered by intraperitoneal injection, seizure-inducing doses were similar for rats with HE and control rats. However, when a constant dose of MPA (330 micrograms) was administered by intracerebroventricular injection, rats with HE took significantly longer to develop seizures than control rats (16.2 vs. 7.3 minutes; p < 0.0005). CONCLUSIONS: In a model of HE: (i) There is increased resistance to the convulsive effects of MPA; and (ii) This phenomenon is apparent when MPA is given centrally, but not when it is given peripherally. Increased resistance to the development of a complication of reduced GABA-mediated neurotransmission induced by MPA in the model provides support for the hypothesis that HE is associated with increased GABA-mediated inhibitory neurotransmission. PMID- 9222688 TI - Urinary excretion of prostaglandins and renal function after hepatic resection. AB - BACKGROUND/AIMS: The leading postoperative complication associated with hepatic resection is the accumulation of fluid in the abdominal cavity, which usually develops approximately one week after surgery. This study was undertaken to investigate the role of renal prostaglandins in modulating renal sodium and water retention in patients who underwent hepatic resection. METHODS: Urinary excretion of thromboxane B2 and 6-keto-prostaglandin F1 alpha as well as renal function were investigated serially in 7 patients with hepatocellular carcinoma who underwent hepatic resection. We administered 60 mg of OKY 046, a selective thromboxane A2 synthetase inhibitor, for 6 hours by continuous drip infusion from the commencement of surgery. RESULTS: Urinary sodium excretion was reduced from 165 mEq/day pre-operatively to 73 mEq/day on postoperative day 6 (p = 0.0181), however, there was no decline in urinary osmorality. OKY 046 administration inhibited intrarenal thromboxane A2 production, on the other hand, it significantly increased the production of intrarenal prostaglandin I2 (from 147 +/- 19 to 6339 +/- 1861 pg/mg creatinine, p = 0.0017) on the day of surgery. The level of thromboxane A2 was significantly increased to 2440 +/- 1099 pg/mg creatinine (p = 0.006, vs. pre-operative value), whereas the level of prostaglandin 12 was significantly reduced to 687 +/- 163 pg/mg creatinine (p = 0.0181, vs. the value on the day of surgery) on postoperative day 6. CONCLUSIONS: Urinary thromboxane A2 synthesis might contribute to sodium and water retention after hepatic resection. These results suggest that combined use of OKY 046 and diuretics prevent ascites formation after hepatic resection. PMID- 9222689 TI - Hepatic dysfunction following cardiac surgery: determinants and consequences. AB - BACKGROUND/AIMS: We prospectively studied the determinants, characteristics, and consequences of hepatic dysfunction in the early postoperative period following cardiac surgery. METHODOLOGY: We examined 3041 adult patients, mean age 60.6 (+/- 8.9), with normal pre-operative liver function who consecutively underwent open heart surgery in a newly established Cardiac Surgery Center. Patients were divided into two groups; Group A included all patients who developed hepatic dysfunction, defined as the presence of jaundice associated with an elevated serum bilirubin above 3 mg/dl, in the early postoperative period. The control group included cardiac surgical patients who did not develop such dysfunction. RESULTS: Hepatic dysfunction developed in 96 patients (3.2%). The affected patients consisted of 63 males and 33 females, mean age 60.8 (+/- 9.4). Determinants of hepatic dysfunction based on univariate analysis were sex, NYHA class, type of surgery, operative times, low cardiac output syndrome necessitating administration of inotropic agents and/or IABP usage, cardiac arrest, presence of hematomas, and number of blood transfusions. Patients with hepatic dysfunction required prolonged mechanical ventilation, stayed longer in the ICU (and in the hospital) and experienced a much higher mortality rate (11.4%) compared to the control group (p = 0.001). CONCLUSION: Although the pathogenesis of hepatic dysfunction seems to be multifactorial, liver cell damage due to decreased perioperative hepatic flow and increased bilirubin load seem to be of critical importance. Early postoperative hepatic dysfunction resulted in increased morbidity and mortality. PMID- 9222690 TI - Lipoperoxide plasma levels in patients with liver cirrhosis. AB - BACKGROUND/AIMS: Oxygen free radicals might play a role in the pathogenesis of tissue damage in many pathological conditions, including liver diseases where antioxidant tissue systems are reduced. The leading mechanism of free radical toxicity is the peroxidation of membrane phospholipids. Lipoperoxide hydrolysis produces aldehydes -the most represented being malondialdehyde-which reacts with thiobarbituric acid and whose concentration is considered a marker of lipid peroxidation. MATERIALS AND METHODS: We developed a fast and cheap HPLC method for measuring malondialdehyde concentration in plasma using a 3 mm C18 Baker column (4.6 x 50 mm). Thiobarbituric acid-reacting substances were eluted isocratically with a mobile phase containing methanol/KH2PO4 (35/65), and detected fluorometrically. The whole analysis lasts 2.5 minutes. The fasting levels of thiobarbituric acid-reactive substances were measured in 30 non-smoking blood donors and in 45 patients with liver cirrhosis. RESULTS: In control subjects they were on average 0.84 [SD 0.41] mumol/L and were increased to 1.59 [SD 1.23] mumol/L in patients with cirrhosis, where they inversely correlated with hepatocellular function. CONCLUSIONS: The method compares favorably with previous techniques in terms of cost and analytical time. It can be used for serial measurement of plasma lipoperoxide concentrations in response to oxidative stress or following drug administration. Our preliminary data confirms the presence of an oxidative stress in cirrhotic patients. Normal lipoperoxide levels in subjects with very advanced disease may be due to polyunsaturated fatty acid deficiency and/or enzyme defects. PMID- 9222691 TI - Enhanced renal susceptibility to ischemia-reperfusion injury in the rat with obstructive jaundice. AB - BACKGROUND/AIMS: To investigate the susceptibility of kidneys to ischemia reperfusion injury under obstructive jaundice. MATERIALS AND METHODS: Bile ducts of male rats were ligated for five days, right kidneys were removed, and vascular clamps were placed across left renal arteries for 30 minutes. RESULTS: Twenty four hours later, ischemia-reperfusion produced renal injury in jaundiced rats as shown by increased serum creatinine and urea nitrogen levels. Histological observation showed tubular damages. These changes were minimal after ischemia reperfusion in control rats. Renal thiobarbituric acid reactive substance contents were increased after bile duct ligation, which did not change after ischemia-reperfusion. On the other hand, ischemia-reperfusion produced a significant increase in renal thiobarbituric acid reactive substance contents in control rats. Renal glutathione contents were increased two fold after bile duct ligation and they were significantly decreased by ischemia-reperfusion. CONCLUSIONS: Kidneys in obstructive jaundice appear to be susceptible to ischemia reperfusion injury. Although protective roles of GSH is suggested, the role of oxygen radicals in this type of renal injury remains to be further elucidated. PMID- 9222694 TI - Hypoplastic right hepatic lobe with retrohepatic gallbladder complicated by hepatolithiasis and liver abscess: a case report. AB - A 37-year-old man with fever, jaundice, severe right upper quadrant abdominal pain and impending septic shock was pre-operatively diagnosed to have hypoplastic right lobe liver with retrohepatic gallbladder, left hepatolithiasis and liver abscess by computed tomography. An urgent operation with cholecystectomy, choledocholithotomy, operative choledochoscopy and T-tube drainage was performed. Postoperative cholangiograms depicted multiple residual stones behind the sharply angulated biliary strictures in the medial branches of the left intrahepatic duct, which could not be eradicated by biliary dilatation via the T-tube tract. The left hepatolithiasis might be coincidental, or secondary to the congenital anomaly of the liver, because of the distorted biliary architecture, and the hypoplastic right lobe liver was associated not only with the retrohepatic gallbladder, but also with the much more complicated left biliary strictures and the hepatolithiasis behind them. PMID- 9222693 TI - Hepatic resection for hepatocellular carcinoma with a tumor thrombus extending to inferior vena cava. AB - The significance of hepatic resection for hepatocellular carcinoma with a tumor thrombus in the inferior vena cava (IVC) is clarified. We operated on 4 patients with HCC who had a tumor thrombus extending to the IVC through the hepatic vein under hepatic vascular exclusion (HVE). In all patients the hepatic resections and thrombectomies were successful without major complication. One patient accompanied with a tumor thrombus in the portal vein had a rash recurrence in the remnant liver and died 6 months after operation. However, three patients without tumor thrombus in the portal vein survived relatively longer post-operatively. Hepatic resection for HCC with tumor thrombus in IVC is acceptable treatment as it is safe. It is considered that better prognoses can be maintained when a tumor thrombus is located only in the hepatic vein, and not in the portal vein. PMID- 9222692 TI - Balloon catheter-assisted inferior vena cava tumor thrombectomy without thoracotomy. AB - We report a technique for tumor thrombectomy of the inferior vena cava for renal cell carcinoma extending into the right atrium that abrogates the need for thoracotomy or sternotomy. PMID- 9222695 TI - Lamivudine treatment of advanced and decompensated liver disease due to hepatitis B. AB - The purpose of this study was to evaluate the effectiveness and safety of lamivudine treatment in patients with advanced and end-stage liver disease caused by hepatitis B. Nine cases of advanced or end-stage liver disease due to hepatitis B infection were treated with lamivudine. Four received liver transplants while receiving lamivudine. Moreover, each of these four has been maintained on lamivudine therapy post-transplantation while receiving immunosuppression. No cases of HBV reactivation have been seen. More importantly, the allograft liver tissue has been HBc and HBs antigen negative as well as HBV DNA negative by PCR. This report suggests that: 1) lamivudine can be given safely to liver transplant candidates; 2) lamivudine suppresses HBV replication, so much so that HBV-DNA becomes undetectable in the serum; 3) despite powerful immunosuppression associated with transplantation, HBV reactivation does not occur under lamivudine therapy; and 4) the observations should cause transplant physicians, surgeons and third-party payers to reconsider their positions relative to transplantation of individuals with HBV-associated cirrhosis. PMID- 9222696 TI - Long-term survival after surgery for advanced intrahepatic cholangiocarcinoma: a case report. AB - Intrahepatic cholangiocarcinoma has a tendency to disperse intrahepatically and extrahepatically, therefore, resectability is limited and prognosis is generally poor. A 68-year-old female was diagnosed as intrahepatic cholangiocarcinoma in the right lobe based on systematic images including computed tomography, magnetic resonance imaging, ultra-sonography, endoscopic retrograde cholangiography, angiography, chest X-ray, as well as laboratory data. Tumor invasion to the right diaphragm, lung and chest wall was suspected pre-operatively. After pre-operative portal embolization, extended right hepatectomy with partial resection of the involved organs including diaphragm, lung, and chest wall was done en bloc. The patient made an uneventful postoperative recovery and there has not been any evidence of recurrence at present, over four and a half years after surgery. Experience in the present case indicates that radical surgery may be a potential approach to yield a hopeful outcome for patient with intrahepatic cholangiocarcinoma, even if the tumor invades adjacent organs directly. PMID- 9222697 TI - Hepatectomy for metastatic renal cell carcinoma. AB - The experience with hepatectomy for metastatic renal cell carcinoma (RCC) has been very rarely reported, because multiple organ metastases ordinarily coexist when hepatic metastases are found out. Three patients who underwent hepatectomy for metastatic RCC are presented here. Radical nephrectomy was performed for the primary renal lesions in all the patients, and their hepatic metastases were resected simultaneously in one of them with a solitary tumor, and about one month later in two of them with multiple (3 and 6) tumors. These operations produced no distinct complications. The patients with 1 and 3 hepatic metastatic lesions survived without tumor recurrence for 12 and 21 months, respectively, while the patient with 6 hepatic metastatic lesions had tumor-free interval of only 2 months and died 10 months after hepatectomy due to lung metastasis. Hepatectomy may be the only promising treatment for hepatic metastases from RCC, but the indication for surgery should be evaluated according to the number of hepatic metastases. PMID- 9222698 TI - The characteristics of hepatocellular carcinoma with a high level of serum lactic dehydrogenase: a case report. AB - Hepatocellular carcinoma (HCC) remains one of the most important malignancies in Japan (1, 2). Lactic dehydrogenase (LDH), which is a glycolyticenzyme, and exists in various types of human tissue and neoplasms, has also been reported to demonstrate a high level (especially LDH 5) in the serum of patients with HCC (3 10). We herein report the findings of a 68-year-old male patient with hepatocellular carcinoma (HCC), whose serum lactic dehydrogenase (LDH) level more closely correlated with the clinical course than the alpha-feto-protein level (AFP). Both the AFP and LDH levels were high before the operation (AFP 1402 ng/ml; LDH 638 IU/L) (LDH1 11%; LDH2 24%; LDH3 34%; LDH4 19%; LDH5 12%). The levels of the serum LDH and AFP one week after the right hepatic lobectomy both decreased to 423 IU/L and 331 ng/ml, respectively. In addition, the LDH isozyme pattern returned to normal. Six weeks after the operation, the serum LDH increased to 3504 IU/L, however, the AFP levels remained low at 43.4 ng/ml, and the CT findings demonstrated multiple recurrent nodules in the whole remnant liver. At eighty-one days after the operation, the patient died due to a rupture of the recurrent HCC. Immunohistochemical observations were performed using the peroxidase labeled streptavidin-biotin technique with slight modifications and using two monoclonal antibodies for AFP and for Ki-67. Most portions of the primary tumor consisted of poorly to undifferentiated HCC. The portion of undifferentiated HCC did not stain for AFP antibody, but the portion of poorly differentiated HCC stained positively for it. It was thus speculated that LDH was mainly produced in the portion of undifferentiated HCC. In addition the undifferentiated HCC were strongly positive for Ki-67, while, in contrast, the poorly HCC was only weakly positive for Ki-67. Based on the above findings, HCC with a high serum level of LDH appears to show both a rapid growth and highly malignant tumors. PMID- 9222699 TI - Surgery for hepatocellular carcinoma with tumor thrombus in the right atrium. AB - Three cases of tumor thrombus that originated from a hepatocellular carcinoma in the liver and extended into the right atrium are described. All patients had received both resection of the tumor thrombus and lobectomy of the liver either simultaneously or independently within a short interval. Surgical order and extracorporeal circulation system were varied depending on the thrombus extension. Two of the patients died within 4 months of surgery due to different reasons and the other is doing well at 24 months after surgery. PMID- 9222700 TI - DNA ploidy, proliferative index and EGF-R status in 130 cases of resected gastric cancer--a multivariate analysis. AB - BACKGROUND/AIMS: The purpose of this study was to define the prognostic role of DNA ploidy, proliferative index and EGF-R status in resected gastric cancer. MATERIALS AND METHODS: Ten clinico-pathological parameters and three biological factors obtained from flow cytometry and immunohisto-chemistry were evaluated in a series of 130 gastric cancer patients who received surgical treatment, including 28 stage IV cases (21.6%), using paraffin-embedded and fresh specimens in 77.7% and 22.3% of the cases, respectively. These variables were first analyzed and tested for correlation within the whole series and then weighted against survival in 117 applicable cases through univariate and multivariate analyses. RESULTS: Aneuploidy was significantly related to higher proliferative activity, EGF-R expression and deeper stomach wall infiltration. Higher proliferative activity was significantly related to deeper stomach wall infiltration and larger tumor diameter. The latter showed a significant relationship to EGF-R expression. Univariate analysis showed the significant variables for survival to be DNA ploidy, pT, pN, M, stage, histological type according to Lauren and tumor diameter. Multivariate analysis calculated on these significant variables using the Cox multiple stepwise regression model detected three factors which independently influence survival: pathological stage (p < 0.00001), histological type according to Lauren (p < 0.002) and DNA ploidy (p < 0.03). CONCLUSIONS: DNA ploidy was shown to be a significant prognostic parameter in resected gastric cancer after pathological stage and histological type according to Lauren. The prognostic roles of proliferative activity and EGF-R status require further investigation. PMID- 9222701 TI - Intra-operative diagnosis of N2 lymph node metastasis of gastric cancer. AB - BACKGROUND/AIMS: Limited lymph node dissection for gastric cancer, which is prevalent in Western countries, leaves cancer cells in the second tier of nodes in patients who have metastasis in those nodes. It is, however, difficult to correctly diagnose nodal status during surgery. The present study was, therefore, designed to examine how to detect N2 metastasis intra-operatively. METHODOLOGY: Five hundred and eight patients undergoing extended lymph node dissections for gastric cancer were retrospectively analyzed. Accuracy of the intraoperative diagnosis of node involvement based on macroscopic findings was investigated, according to the N stage and histological type of the tumor. Furthermore, the distributions of N2 metastasis were clarified, according to tumor site. RESULTS: Intra-operative macroscopic findings were frequently assessed as being less severe than histological findings in cases with N2 metastasis (61.9%, 39/63). Intra-operative recognition of N2 metastasis was significantly lower in the cases with undifferentiated adenocarcinoma (28.2%, 11/39) than in those with differentiated adenocarcinoma (56.5%, 13/23). The distributions of N2 metastasis revealed nodes along the left gastric and common hepatic arteries to be the key junctions for lymphatic flow from the middle and lower thirds of the stomach, respectively. CONCLUSIONS: Intra-operative diagnosis of N2 metastasis is difficult to make based on macroscopic findings, especially in undifferentiated tumors. To detect N2 metastasis intra-operatively, the nodes along the left gastric or common hepatic artery should be submitted to frozen section examination for primary tumors located in the middle or lower third of the stomach, respectively. PMID- 9222703 TI - Helical CT of the abdomen after pancreaticoduodenectomy: usefulness for detecting postoperative complications. AB - BACKGROUND/AIMS: Helical (spiral) computed tomography (CT) decreases scanning time and respiratory motion artifact. These characteristics are convenient for the evaluation of postoperative intra-abdominal conditions. We investigated the usefulness of helical CT in detecting complications after pancreaticoduodenectomy. MATERIAL AND METHODS: Helical CT scans were carried out in 21 patients with symptoms suggesting postoperative complications after pancreaticoduodenectomy. Twenty-one patients who underwent pancreaticoduodenectomy were examined postoperatively by helical CT scans. Findings were correlated with the subsequent clinical course. RESULTS: Abnormal findings were detected by helical CT scan in 16 (76%) of 21 patients. These included 13 patients with fluid collection, 7-with localized liver infarction, 5 with intra-abdominal abscess, 3 with pleural effusion or ascites, and 2 with pseudoaneurysms or wound abscesses. Liver infarctions were found in 7 of 10 patients who underwent combined resection of the portal and/or superior mesenteric veins, but not in any of 11 patients without combined vascular resection. CONCLUSIONS: Helical CT was useful in evaluating intra-abdominal conditions and detecting complications after pancreaticoduodenectomy. PMID- 9222702 TI - Chronic calcifying pancreatitis in Taiwan: a multicentric study and comparison with western countries. AB - BACKGROUND/AIMS: This study investigates the clinical features of chronic calcifying pancreatitis (CCP) in Taiwan and also the comparative differences in the disorder as it affects orientals and occidentals. MATERIALS AND METHODS: Medical records at seven tertiary hospitals relating to patients diagnosed with CCP between 1976 and 1996 are reviewed and analyzed. Ninety patients were enrolled. Defining the calcification of the pancreas is achieved by plain film, ultrasonography, computed tomography, or histology. RESULTS: CCP afflicts men more frequently than it does women, by a ratio of 3.5:1 (70 men and 20 women). The mean age is 45 years (male: 46 female: 41.4). For fifty-two patients (57.8%), alcohol is the major cause of the condition, while in others, the causes are non alcoholic (idiopathic: 31; biliary: 4; hereditary: 3). Alcoholism is mainly associated with males and younger sufferers. The major complications are diabetes mellitus (53.3%), cysts or pseudocysts (21.1%), and biliary stricture or stones (20%). Pancreatic adenocarcinoma and splenic vein thrombosis were found in six and five patients, respectively. Three patients died from cancers of other than pancreatic origin (lung: 1;liver: 1;bile duct: 1). Thirty-three patients were treated surgically of which thirteen (39.4%), including one with pancreatic auto transplantation, improved. Fifty-seven patients received medical treatment but only eleven (19.3%) improved. CONCLUSIONS: The clinical features of CCP in Taiwan are notably similar to those manifesting in western countries and in Japan. With the changes in life style and increased alcoholic consumption in Taiwan, the prevalence of CCP may increase and its demographic features may alter in the future. PMID- 9222704 TI - Postoperative delayed emptying in pylorus-preserving pancreatoduodenectomy using pancreaticogastrostomy: comparison of the reconstruction position. AB - BACKGROUND/AIMS: We frequently use pancreaticogastrostomy after pylorus preserving pancreatoduodenectomy (PpPD). Although it is a useful and effective technique, it sometimes requires prolonged delayed emptying. We considered the best position for pancreaticogastrostomy in such a way as to reduce this period of delayed emptying. MATERIAL AND METHOD: From 1991 to 1996, 17 patients whose blood sugar was controlled and who were kept on H2 blocker were selected from among 25 patients who underwent PpPD and were reconstructed by pancreaticogastrostomy. We investigated the history of the relationship between reconstruction position and delayed emptying, reconstruction position and minor leakage, minor leakage and delayed emptying, reconstruction position and stomach mobility without leakage. RESULTS: The delayed emptying was shorter for a reconstruction position in the area above the angulus rather than below it. CONCLUSION: The results suggest that the area above the angulus is the preferred site for pancreaticogastrostomy after PpPD. PMID- 9222705 TI - How efficient is endoscopic injection sclerotherapy in peptic ulcer hemorrhage. AB - BACKGROUND/AIMS: Peptic ulcer hemorrhage is a common, worldwide problem and a major cause of morbidity and mortality. The aim of this study was to establish the percentage of patients with bleeding peptic ulcers who were treated surgically because endoscopy failed to stop the hemorrhage. METHODOLOGY: This retrospective analysis includes patients from our institution who underwent urgent endoscopic examination of the upper digestive tract and hemostatic interventions with injection therapy (sol. 1:10000 adrenaline and 1% polidocanol) between January, 1994 and November, 1995. RESULTS: Two hundred thirty-three patients with bleeding peptic ulcers were examined: 111 with bleeding gastric ulcers (66 males, 45 females; average age 60.21 years, SD +/- 13.94; span 28-94 years) and 122 with bleeding duodenal ulcers (95 males, 27 females; average age 55.24 years, SD +/- 17.35; span 16-88 years). In all patients, injection sclerotherapy was performed. The ulcers were classified according to Forrest's classification of bleeding activity. In 10 patients (4.2%) with acute hemorrhage (6 males; average age 63.2 years, SD +/- 5.6; span 53-70 years: 4 females, average age 61.0 years, SD +/- 11.82; span 51-81 years), endoscopic hemostasis did not prove successful and they were treated operatively. In 5 cases, the cause of hemorrhage was a gastric ulcer and in 5 others, duodenal ulcer. During the postoperative period, 5 patients died of complications. CONCLUSIONS: Endoscopic hemostasis has been a major therapeutic advancement in the management of peptic ulcer hemorrhage and has influenced surgical management. Injection sclerotherapy is a low cost, effective and safe procedure which is easy to implement in a variety of clinical settings. Early elective operation after initial endoscopic hemostasis is the wisest choice for elderly patients with co-existing disease and selected patients at high risk for recurrent bleeding. PMID- 9222706 TI - Pre-operative serum levels of CA72-4 in patients with gastric adenocarcinoma. AB - BACKGROUND/AIMS: A retrospective study was designed to evaluate the clinical significance of preoperative serum levels of carcino-embryonic antigen (CEA) and CA72-4 in patients with advanced gastric adenocarcinoma. METHODOLOGY: Pre operative serum levels of CEA and CA72-4 were analyzed and compared with the proliferative activity of cancer cells, monitored in terms of the number of argyrophilic nucleolar organizer regions (AgNORs), in tumors from 153 patients with resected gastric adenocarcinoma with serosal invasion. RESULTS: Elevated levels of CEA were frequently found in patients with lymph node metastasis and liver metastasis. By contrast, elevated levels of CA72-4 were frequently found in patients with peritoneal metastasis at the time of operation. A close correlation was found between pre-operative levels of serum CA72-4 and the AgNOR scores of tumors. Even when apparently curative operations were performed, peritoneal metastasis occurred these were detected quite soon, after apparently curative operations in patients with high pre-operative serum levels of CA72-4 and tumors with high proliferative activity. CONCLUSIONS: This phenomenon might explain the poor prognosis of patients with high levels of CA72-4. PMID- 9222707 TI - Effect of gastric mucosal protection in nonrheumatoid patients with short-term nonsteroidal anti-inflammatory drug therapy: a prospective randomized multicenter study. AB - BACKGROUND/AIMS: Since nonsteroidal anti-inflammatory drugs (NSAIDs) are known to cause gastrointestinal mucosal injury, concurrent administration of gastric protective agents in patients with rheumatoid disorders is often recommended. The present study was designed to determine the possible benefit of sucralfate, a gastric protective agent in patients receiving short-term therapy with NSAIDs for conditions other than rheumatoid arthritis. METHODOLOGY: In nineteen general practitioners offices a total of 208 patients were studied. Patients received either sucralfate suspension (2 x 2 g/day) or no additional medicine while being put on NSAIDs for a mean of 9 days. RESULTS: Forty patients (39%) in the sucralfate group and 71 (68%) in the control group developed gastrointestinal symptoms (p < 0.05). In the control group six patients (6%) had gastrointestinal hemorrhage and 2 patients (2%) experienced perforation of a duodenal ulcer. Gastrointestinal hemorrhage and perforation were not observed in the sucralfate group. Short-term NSAID use in nonrheumatoid patients carries a higher risk for symptoms and complications in patients that are 40 years old and above. These risk are even more pronounced in patients of 60 years of age or above, in patients with a history of peptic ulcer disease, in smokers, in patients with concomitant disease and concomitant steroid use, and with the use of diclofenac. The mean age of patients with gastrointestinal hemorrhage was 65 years. Both patients that suffered perforation of a duodenal ulcer were young (35 and 42 years) and had a history of duodenal ulcer disease. CONCLUSION: Gastric protection with sucralfate reduces the frequency of both symptoms and complications in nonrheumatoid patients receiving short-term therapy with NSAIDs. PMID- 9222708 TI - Cellular proliferation and ploidy of the gastric mucosa: the role of Helicobacter pylori. AB - BACKGROUND/AIMS: Recently, H. pylori has been recognized as a risk factor for gastric adenocarcinoma. As such, we have analyzed the DNA content of gastric epithelial cells in an attempt to reveal the role of H. pylori in gastric carcinogenesis. METHODOLOGY: Fifty-three subjects presented with gastric dyspepsia, 39 males and 14 females, with a mean age of 42.15 (+/- 13.16) years. They were referred to the out-patient clinic to undergo endoscopic examination for the first time. Biopsy specimens from the antrum of each subject were subjected to culture for the presence of H. pylori histologic diagnosis, and DNA flow cytometry for the analysis of cellular proliferation and DNA policy. RESULTS: The endoscopic diagnoses were normal appearance (12), Gastric ulcer (12), duodenal ulcer (29). Thirty-eight (72%) subjects were positive, and 15 (28%) subjects were negative for H. pylori. Abnormal DNA-content (aneuploidy) was found in specimens from the antrums of 3 patients, 2 patients with duodenal ulcers (DU, and one with a gastric ulcer (GU). The cellular proliferation detected by flow cytometry in the form of proliferative index (PI; percentage of cells in the DNA S and G2M phases) was 27.88 (+/- 12.48) and 14.17 (+/-2.94) in the antrums of those positive and negative for H. pylori, respectively. A very significant increase in the PI (p < 0.005) was found between subjects positive and negative for H. pylori. Patients with DU and H pylori infection had the highest PI, and the PI was significantly higher than in patients with DU, but without infection. Regarding histology, there was a significant increase in the PI in the presence of H. pylori infection in either CAG or dysplasia groups as compared to cases without infection in the same groups. CONCLUSION: These results show that H. pylori infection is associated with changes in the DNA-content and cellular proliferative activity, suggesting that H. pylori may be implicated in gastric carcinogenesis. Also, the significant increase in the PI along the progression of severity of the disease suggests that measuring this parameter might allow more accurate monitoring of patients, so that a targeted therapeutic protocol may be defined. PMID- 9222709 TI - Pantoprazole-based dual and triple therapy for the eradication of Helicobacter pylori infection: a randomized controlled trial. AB - BACKGROUND/AIMS: The eradication of Helicobacter pylori (Hp) infection in duodenal ulcer and dyspepsia has been achieved using various therapy regimens. The efficacy of protein pump inhibitor pantoprazole as part of these regimens has not been widely studied. METHODOLOGY: During a prospective randomized trial, 250 Hp positive patients with either duodenal ulcer, erosive bulbitis, or gastritis and dyspepsia were treated using 14 days of therapy 1) pantoprazole 40 mg daily and clarithromycin 500 mg b.i.d. (PC), 2) pantoprazole 40 mg daily and clarithromycin 500 mg b.i.d. plus amoxicillin 1 g b.i.d. (PCA), or 3) bismuth subcitrate 120 mg t.i.d., roxithromycin 150 mg b.i.d., metronidazole 250 mg b.i.d. plus ranitidin 300 mg (BRMR). Hp status was assessed on 3 tests at the inclusion (2-specimen rapid urease test, 2-specimen histology, serology) and 2 tests (2-specimen rapid urease test, 2-specimen histology) 4 weeks after the end of the treatment. RESULTS: The entry criteria was fulfilled in 250 patients, of whom 13 missed the control endoscopy. The treatment had to be discontinued for adverse effects in 8 (10%) BRMR patients, and 1 (1%) PCA patients. Compliance was 100% in the PC group. All ulcers were healed at the end of the study with one exception in the BRMR group. The best eradication rate of Hp was shown by the PCA group with 94.8% (n = 73/77) followed by the PC group with 82.5% (n = 66/80) and finally the BRMR with 67.6% (n = 48/71)-PCA:BRMR - p < 0.001; PC:BRMR-p < 0.001; PCA:PC-p < 0.05. CONCLUSION: This study showed that triple therapy using PPI pantoprazole combined with antibiotics clarithromycin and amoxicillin was very effective in the eradication of Hp and treatment of duodenal ulcer with rare side effects. The dual pantoprazole and clarithromycin therapy had the highest rate of patient compliance, but is less effective than triple therapy. The combination of ranitidin with bismuth based triple therapy had the highest number of adverse events and the lowest rate of Hp eradication and therefore, should not be recommended. PMID- 9222711 TI - Comparison in survival between hepatic metastases of gastric and colorectal cancers. AB - BACKGROUND/AIM: The outcome after hepatectomy and non-surgical treatment of liver metastases from gastric and colorectal malignancies are reported. METHODOLOGY: Between April 1988 and March 1994, 176 patients with metastatic liver cancer were treated at the First Department of Surgery, Kyoto Prefectural University of Medicine Hospital. RESULTS: All patients received multi-disciplinary treatment, and 51 underwent hepatectomy. The survival after hepatectomy for metastatic liver cancer from a colorectal primary was better than that for gastric cancer. The survival after hepatic arterial infusion (HAI) therapy for metastases from gastric cancer was better than that for colorectal cancer. CONCLUSION: Surgical resection may be the best treatment for liver metastases from colorectal cancer. HAI may be a better option for liver metastases from gastric cancer. PMID- 9222710 TI - Jejunal pouch length influences metabolism after total gastrectomy. AB - BACKGROUND/AIMS: To determine whether the length of the jejunal pouch used for reconstruction after total gastrectomy influences the postoperative metabolic and clinical outcome. METHODOLOGY: Twenty-one patients who underwent total gastrectomy with long jejunal pouch reconstruction (length 20 cm: n = 9) or short jejunal pouch reconstruction (length 15 cm: n = 12) were studied. The volume of a single meal, clinical symptoms, glucose metabolism, and other metabolic parameters were studied. RESULTS: Food intake, body weight, and dumping symptoms were similar in both groups, but the short pouch group had less reflux symptoms. Serum albumin and total cholesterol levels were higher in the short pouch group than in the long pouch group at 12 months postoperatively (p < 0.05). However, the short pouch group showed reduced glucose tolerance. CONCLUSION: These findings suggest that jejunal pouch length might have a role in postoperative physiological status. PMID- 9222712 TI - Emptying of the jejunal pouch as a gastric substitute after total gastrectomy for cancer. AB - BACKGROUND/AIMS: The reservoir and transit capacity of the post-gastrectomy jejunal pouch was evaluated, using a radioisotopic method, to examine the relationship of the gastric emptying to postprandial symptoms and to the food intake status. METHODOLOGY: Thirty-seven patients who had undergone total gastrectomy for cancer (Roux-Y reconstruction, 8; Hunt- Lawrence pouch and Roux Y, 15; pouch interposition, 5; modified pouch interposition, 9) were retrospectively studied. Based on the percent retention in the gastric substitute, the emptying curves were classed as showing delayed, intermediate, and rapid emptying types. RESULTS: All of the patients with pouch reconstruction showed either delayed or intermediate emptying. High frequency of the sensation of epigastric fullness, nausea, or vomiting was demonstrated in the patients with delayed emptying (p < 0.01). The patients with delayed emptying showed poor food intake compared to those with intermediate emptying (p < 0.05). Given the X-ray video film and endoscopic findings, the delayed emptying was thought to be due to poor drainage of the efferent loop resulting from the post-operative adhesions. CONCLUSIONS: The present study revealed that delayed emptying is associated with postprandial symptoms and with poor food intake. The examination of gastric emptying is useful in evaluating or predicting the postoperative status. PMID- 9222713 TI - Surgical outcome in early gastric cancer with lymph node metastasis. AB - BACKGROUND/AIMS: Definitive surgical management of early gastric cancer with lymph node metastasis has not been established. This paper describes the clinico pathologic characteristics of early gastric cancer with lymph node metastasis. MATERIALS AND METHODS: A retrospective study of early gastric cancer with lymph node metastasis (32 patients) was performed to compare clinico-pathologic features with patients without lymph node metastasis (283 patients). RESULTS: All patients with lymph node metastasis had submucosal gastric invasion. The incidence of histologically proven curative resection in patients with lymph node metastasis was significantly lower than in those without metastasis (40.6% versus 93.3%). The 5-year survival rate was poorer in patients with positive nodes than in those with negative nodes (83.8% versus 96.2%). Recurrence was more frequent in patients with involved nodes (12.5% versus 0.4%). Lymph node metastasis was more frequent with the following: submucosal invasion, tumor over 5 cm in size, positive venous involvement, and an advanced growth pattern. CONCLUSIONS: Pre operative and intra-operative evaluation for lymph node metastasis is essential for the appropriate surgical treatment of early gastric cancer. PMID- 9222714 TI - Laparoscopic re-operation from gastro-oesophageal reflux. AB - Since 1994 until the present day, we have had to surgically re-operate in five cases of failure with laparoscopic operations aimed at correcting gastro oesophageal reflux disease. Two of these cases came from our own patients and three came under our observation from other centers. We applied fundoplication according to Nissen-Rossetti in three cases and the Rossetti-Hell operation in the other cases. One case involved recurrent gastro-oesophageal reflux with a short oesophagus and fundoplication raised into the mediastinum. In one other case, there was recurrent hiatal herniation with a rotary as well as axial component and consequent mediastinal occupation. The other three cases featured persistent post-operative dysphagia caused, in one case, by an error in the creation of the anti-reflux valve (perigastric cuff) and, in the other two, by erroneous choice of the anti-reflux operation: post-operative manometry showed important oesophageal hypo-dyskinesia which should have indicated partial fundoplication. All the patients underwent laparoscopic exploration. The patient with the short oesophagus had to be converted for the performance of a total duodenal diversion, while the remaining four patients underwent a total laparoscopic operation. The patient with recurrent hiatal hernia had the hernia reduced in the abdomen and combined anterior and posterior hiatoplasty. In another three cases, total fundoplication was transformed into partial fundoplication according to Toupet. The post-operative course and clinical results were excellent in all five patients. Stress is placed on the importance of accurate morphological and functional assessment of the oesophagus in the pre operative stage so as to select the most suitable operation and in the post operative stage in order to evaluate the causes of failure, the advantages of laparoscopy in terms of exposure of the operative field, the importance of certain technical details that optimize the results of the operation, and the efficacy of the laparoscopic approach also for the correction of most failures that demand re-operation. PMID- 9222716 TI - IEFS pan-EU survey of consumer attitudes to food, nutrition and health. PMID- 9222715 TI - Regression of gastric polyps in Gardner's syndrome with use of indomethacin suppositories: a case report. AB - Patients with familial adenomatous polyposis most often have multiple polyps in the large bowel and also in the gastroduodenal region. There are reports of regression or disappearance of colorectal and duodenal polyps while on sulindac therapy. We report here what seems to be the first case of regression of gastric polyps while on indomethacin suppository treatment. PMID- 9222717 TI - Methods used to conduct the survey on consumer attitudes to food, nutrition and health on nationally representative samples of adults from each member state of the European Union. PMID- 9222718 TI - Influences on food choice perceived to be important by nationally-representative samples of adults in the European Union. AB - OBJECTIVE: The purpose of this baseline survey was to obtain comparable data on perceived influences on food choice from EU member countries as the starting point for EU healthy eating promotion campaigns and programmes. DESIGN: A cross sectional study in which quota-controlled, nationally-representative samples of approximately 1000 adults from each country completed a face-to-face interview assisted questionnaire. SETTING: The survey was conducted between October 1995 and February 1996 in the 15 member states of the European Union. SUBJECTS: 14331 subjects (aged 15 y upwards) completed the questionnaire. Data were weighted by population size for each country and by sex, age and regional distribution within each member state. RESULTS: The five most important factors influencing consumers food choice were 'quality or freshness' (74%), 'price' (43%), 'taste' (38%), 'trying to eat healthy' (32%) and 'family preferences' (29%). Subjects in different categories (age, sex, education and employment status) selected different factors as having major influence on their food choice. Demographic factors seemed to have greater effects on perceived influences than culture (country): 'quality/freshness', 'price', 'trying to eat healthy', 'family preferences' seemed to be most important in women, 'taste' and 'habit' in males. Females and older and more educated subjects were more likely than other subjects to select 'trying to eat healthy' as having a major influence. 'Price' seemed most important in unemployed and retired subjects. PMID- 9222719 TI - Sources used and trusted by nationally-representative adults in the European Union for information on healthy eating. AB - OBJECTIVE: To assess what sources of information on healthy eating are used and in particular to ascertain which are the most trusted by European adults. DESIGN: A cross-sectional study in which quota-controlled, nationally-representative samples of approximately 1000 adults from each country completed a face-to-face interview-assisted questionnaire. SETTING: The survey was conducted between October 1995 and February 1996 in the 15 member states of the European Union. SUBJECTS: 14331 subjects (aged 15 y upwards) completed the questionnaire. Data were weighted by population size for each country and by sex, age and regional distribution within each member state. RESULTS: The five sources of information most frequently selected were: TV/radio (29%), magazines and newspapers (27%), health professionals (26%), food packages (22%) and relatives/friends (22%). Those used by less than 5% of the population included vegetarian and slimming societies and women's organizations. The reliance on health professionals for information was stronger for females and tended to increase with age. The most trusted sources of information in almost all countries were health professionals (91%) and government agencies (80%) with great consistency across countries. 15% of Europeans stated that they did not get any information on healthy eating. PMID- 9222720 TI - Definitions of 'healthy' eating: a pan-EU survey of consumer attitudes to food, nutrition and health. AB - OBJECTIVE: To describe the perceptions of a healthy diet across Europe and to explore the socio-cultural factors that influence these perceptions. DESIGN: A cross-sectional study in which quota-controlled, nationally-representative samples of approximately 1000 adults from each country completed a face-to-face interview-assisted questionnaire. SETTING: The survey was conducted between October 1995 and February 1996 in the 15 member states of the European Union. SUBJECTS: 14331 subjects (aged 15 y upwards) completed the questionnaire. Data were weighted by population size for each country and by sex, age and regional distribution within each member state. RESULTS: Responses were grouped into broad categories; overall 80% (specific country rates varied from 67-91%) of respondents mentioned either more fruit and vegetables or less fat, fatty foods, or a low fat diet, or balance and variety. The effects of age, gender and level of education were also explored: educational level appeared to have the strongest influence on perceptions of a healthy diet. Respondents who mentioned the family as a key influence on food choice, were more likely to mention eating more fruit and vegetables as part of a healthy diet. Respondents who stated that they did not have any source of information about diet were less likely to mention balance and variety or less fat or more vegetables. CONCLUSIONS: The results of the present study suggest that many people defined healthy eating in a way which would suggest that the healthy dietary guidelines are having some impact. The results also show, however, that there may be specific groups who are missed by current national campaigns, and that any European wide campaigns to change attitudes about healthy eating need to consider the baseline perception of healthy eating reported here. PMID- 9222721 TI - Perceived need to alter eating habits among representative samples of adults from all member states of the European Union. AB - OBJECTIVE: To examine the perceived need to alter eating habits among nationally representative samples from each member state of the European Union (EU). DESIGN: A cross-sectional study in which quota-controlled, nationally-representative samples of approximately 1000 adults from each country completed a face-to-face interview-assisted questionnaire. SETTING: The survey was conducted between October 1995 and February 1996 in the 15 member states of the European Union. SUBJECTS: 14,331 subjects (aged 15 y upwards) completed the questionnaire. Data were weighted by population size for each country and by sex, age and regional distribution within each member state. RESULTS: 71% of EU subjects agreed with the statement 'I do not need to make changes to the food I eat, as it is already healthy enough'. There was wide variation between the member states ranging from 47% in Finland to 87% in Italy indicating agreement. Overall there was little difference between the sexes except in Austria, Belgium, Germany, Greece and Ireland, but the proportions of subjects agreeing with the statement generally increased with advancing age and decreased with higher levels of education. The effects of demographics were not consistent across member states. A total of 49% of EU subjects agreed with the statement 'I usually do not think of the nutritional aspects of the food I eat'. Significantly more females than males disagreed with the statement in all countries except Portugal. In all member states there were widespread beliefs that people in general should decrease their consumption of savoury snacks and increase their consumption of fruit and vegetables. CONCLUSIONS: The results of this study demonstrate that dietary advice may not be perceived as personally relevant among EU subjects. In addition important target groups for the promotion of healthy eating have been identified for example, males or subjects with low levels of education. Because of the variation in attitudes a single pan-EU healthy eating programme is unlikely to be effective for all countries or for different demographic groups. PMID- 9222722 TI - Difficulties in trying to eat healthier: descriptive analysis of perceived barriers for healthy eating. AB - OBJECTIVE: To determine the factors which are perceived to be important barriers to healthy eating among European adults. DESIGN: A cross sectional study in which quota-controlled, nationally-representative samples of approximately 1000 adults from each country completed a face-to-face interview-assisted questionnaire. SETTING: The survey was conducted between October 1995 and February 1996 in the 15 member states of the European Union. SUBJECTS: 14,331 subjects (aged 15 y upwards) completed the questionnaire. Data were weighted by population size for each country and by sex, age and regional distribution within each member state. RESULTS: The study demonstrates a great variability in the perceived barriers to healthy eating between different EU countries. Lack of time was the most frequently mentioned difficulty among EU subjects for not following nutritional advice (24% of total EU sample). This barrier was frequently reported by the younger and the higher education people. Other frequently reported barriers were giving up favourite foods (23%) and willpower (18%). Thus healthy diets do not appear to be viewed as an easy or attractive alternative to current diets. There was wide geographical variation in the number of subjects mentioning price as an important barrier to healthy eating (15% in overall EU sample) ranging from less than 10% in Germany and Italy to 23% in the UK and 24% in Luxembourg. PMID- 9222723 TI - Perceived benefits of healthy eating among a nationally-representative sample of adults in the European Union. AB - OBJECTIVE: To determine the main perceived benefits associated with healthy eating among European adults. Efforts to make a healthy diet more attractive have to be based on motives capable of stimulating alterations in nutritional behaviour. DESIGN: A cross-sectional study in which quota-controlled, nationally representative samples of approximately 1000 adults from each country completed a face-to-face interview-assisted questionnaire. SETTING: The survey was conducted between October 1995 and February 1996 in the 15 member states of the European Union. SUBJECTS: A total of 14331 subjects (aged 15 y upwards) completed the questionnaire. Data were weighted by population size for each country and by sex, age and regional distribution within each member state. RESULTS: The question 'which specific benefits, if any, would you personally believe can be achieved by healthy eating'? was answered from a collection of 9 given items with stay healthy by 67% of subjects, with prevent disease by 66%, with be fit by 53%, with control weight by 53% and with quality of life by 45%. The most important personal benefit was asked by the question 'which one benefit would be the most personally significant for you'? 31% of subjects stated stay healthy, 24% prevent disease, 10% control weight, 10% quality of life, 9% be fit. The frequency of answered statements differed considerably between the EU countries, however, no regional structure was detected. More women then men expected benefits from healthy eating. With increasing age more people tended to believe in the benefits stay healthy and prevent disease. Be fit was a more important benefit for younger people (aged 15-34 y). People with a higher level of education associated more benefits from healthy eating. PMID- 9222724 TI - Stages of dietary change among nationally-representative samples of adults in the European Union. AB - OBJECTIVE: To investigate the distribution across the different stages of change for each of the 15 participating European countries, and the effect of socio demographic variables such as sex and education on this distribution. Also to assess the relationships between stages of change and influences of food choice, and other variables. DESIGN: A cross-sectional study in which quota-controlled, nationally-representative samples of approximately 1000 adults from each country completed a face-to-face interview-assisted questionnaire. SETTING: The survey was conducted between October 1995 and February 1996 in the 15 member states of the European Union. SUBJECTS: 14331 subjects (aged 15 y upwards) completed the questionnaire. Data were weighted by population size for each country and by sex, age and regional distribution within each member state. Subjects were divided into five different categories according to their attitudes towards 'changing their eating habits in order to eat healthier': (1) Precontemplation; do not consider any changes, (2) Contemplation; consider changes, (3) Decision; make plans to change, (4) Action; carry out the changes, and (5) Maintenance; maintained changes for more than six months. RESULTS: 52% of the subjects were in the precontemplation stage, whereas 31% of the subjects were in the maintenance stage. Two, one, and seven percent of subjects were in the contemplation, decision and action stage, respectively. In the Mediterranean countries, and in Germany, there were more people (55-64%) in the precontemplation stage, whereas in the Scandinavian countries there were less people in precontemplation stage (20-38%). The opposite was true for the maintenance stage, whereas women and people with a higher education level tended to be more in the maintenance stage. With respect to influence on food choice, subjects in precontemplation stage found that taste was more important, whereas people in maintenance stage found that health was more important. CONCLUSIONS: The stages of change model makes a useful distinction between people with different attitudes towards nutrition and health. Nutrition education can benefit from this distinction. PMID- 9222725 TI - The intestinal effects of bran-like plastic particles: is the concept of 'roughage' valid after all? AB - OBJECTIVE: The mechanisms by which dietary fibre exerts is laxative action are not fully understood. Studies using sliced plastic tubing as a fibre substitute showed a decrease in both small and large bowel transit time. The significance of these studies is hard to interpret. We set out to compare the effects on intestinal function of wheat bran with plastic flakes similar in size and flaky shape to wheat bran (and devoid of plasticizers). DESIGN AND METHODS: Volunteers consumed coarse wheat bran then, after a washout period, plastic flakes of the same size and shape as the bran. Before and after each intervention whole-gut transit time (WGTT), defecation frequency, stool form, stool water content, stool beta-glucuronidase activity and dietary intake were assessed. RESULTS: Twenty nine volunteers consumed a mean of 27.1 g of raw wheat bran and 24 g of plastic flakes a day. Baseline WGTT, interdefecatory intervals (IDI), stool form, weight, output, water content, and beta-glucuronidase were similar before both interventions. Both led to a decrease in mean faecal beta-glucuronidase activity, median WGTT (bran 25.8%, plastic 28.6%) and IDI (bran 23.3% plastic 25.0%). Both also increased stool form score (bran 28.6%, plastic 21.2%) and stool output (bran 67.1%, plastic 79.0%). Stool water content only rose with wheat bran (72% 75%, P = 0.014). CONCLUSION: Overall, plastic 'pseudobran' was as effective at altering colonic function as wheat bran at a similar dosage but with fewer particles. The mechanism is not by increased faecal water. Reduction in enzyme activity with plastic flakes suggests that the plastic led to qualitative and, probably, beneficial changes in the bacterial flora or their metabolic processes. The concept of roughage deserves to be revived. PMID- 9222726 TI - Association between serum levels of total IgA and IgA class endomysial and antigliadin antibodies: implications for coeliac disease screening. AB - BACKGROUND: Patients with selective immunoglobulin A (IgA) deficiency and coeliac disease, an established association, lack serum IgA class antigliadin and endomysial antibodies (AGA, EmA). Diagnostic protocols relying on AGA and EmA to select patients for small bowel biopsies will not identify these patients. OBJECTIVE: To determine whether total IgA should be routinely measured in patients, suspected of having coeliac disease as a supplementary screening test before biopsy. DESIGN: Prospective measurement of IgA, AGA and EmA in patients undergoing small bowel biopsy for suspected coeliac disease. PATIENTS: We studied 318 patients suspected of having coeliac disease. Sera from 1959 controls in a random population sample were assayed as controls. RESULTS: Thirty-one (10%) patients had villous atrophy, of whom 27 (87%) had EmA. Five (2%) of the 318 patients had undetectable total IgA (< 0.07 g/l): two (40%) of these five had villous atrophy in the setting of negative EmA. Use of undetectable IgA as a selection criterion for small bowel biopsy as well as positive EmA would have improved sensitivity from 87% (27/31) for EmA alone to 94% (29/31), with a fall in positive predictive value from 100% (27/27) to 91% (29/32), but would have maintained high specificity and negative predictive value. Serum IgA was undetectable in 5 (4%) of 117 patients with AGA in the range 0-10 ELISA units (EU) compared with none of 201 with higher AGA (P = 0.007, Fisher's exact test). Compared with controls who had AGA 0-10 EU, patients were more likely to have undetectable IgA (5/117 (4%) vs. 3/706 (0.4%); P = 0.005). Overall, the median IgA in patients with AGA 0-10 EU was lower than for those with AGA > 10 EU (1.89 g/l, vs. 2.34 g/l, P < 0.001). CONCLUSION: There is an association between IgA deficiency and low/negative EmA/AGA. Routine measurement of total serum IgA in patients suspected of having coeliac disease, either with EmA or where AGA is low, improves selection of patients for small bowel biopsy. PMID- 9222727 TI - Laser fragmentation of pancreatic duct stones using a rhodamine laser with an automatic stone-tissue detection system. Basic in-vitro studies. AB - OBJECTIVES: The aim of our study was to examine the suitability of a rhodamine 6G laser with an integrated stone-tissue detection system (STDS) for fragmenting pancreatic stones. METHODS: A total of 64 pancreatic duct stones were measured for weight, diameter, main chemical components and in some cases for their computerized tomography density. Recognition of all stones was checked with the standard STDS or a prototype version. Number of fragmentation pulses, total fragmentation energies and correlation with the individual stone parameters were determined. The quality of the tissue-detection mode was evaluated in postmortem pancreata. RESULTS: The standard STDS detected only 45% of the pancreatic stones correctly. When using the prototype, the detection rate improved significantly up to 75% (P < 0.01). All laser pulses were cut off if tissue contact occurred. All the stones were completely disintegrated by the laser pulses. A slight correlation was found only between the required fragmentation energy and the stone weight (linear regression: R2 = 0.77); other factors had no significant impact. CONCLUSION: The rhodamine 6G laser is suitable for the fragmentation of pancreatic stones in vitro. The integrated STDS is less effective for pancreatic stones than reported for biliary stones. PMID- 9222728 TI - Santorini's duct--risk factor for acute pancreatitis or protective morphologic variant? Experiments in rabbits. AB - BACKGROUND AND AIMS: Gallstone pancreatitis is assumed to result from stone passage through the choledochoduodenal junction. Stone impactions may either result in the obstruction of the pancreatic duct or occur below the confluence of the biliary tract and the pancreatic duct and, thus, may favour bile reflux into the pancreatic duct. We studied effects of a patent Santorini's duct upon secretory flow and pancreas morphology under both conditions. METHODS: A catheter in the distal rabbit pancreatic duct created a second outlet for pancreatic juice and, thus, mimicked a patent Santorini's duct. A second catheter was introduced into the proximal pancreatic duct and into the common bile duct. This catheter mimicked a common channel behind a papillary obstruction. Clamping of this catheter mimicked a stone obstruction of the pancreatic duct. A catheter in the cystic duct allowed for the infection of bile with 10(7) Escherichia coli bacteria/ml. The flow direction of bile and pancreatic juice was directly observed. Pancreatic histology was analysed after 24 h. RESULTS: Pancreatic duct obstruction produced an oedema of the gland. Creation of a patent Santorini's duct prevented development of the histological changes caused by pancreatic duct obstruction. In rabbits in which a common channel obstruction was mimicked, Santorini's duct produced flow of bile along the pancreatic duct system. Flow of sterile bile along the duct did not cause pancreatic inflammatory lesions. Bile that was infected with E. coli bacteria produced an acute interstitial-oedematous pancreatitis. CONCLUSIONS: (1) A patient Santorini's duct protects the gland from the effects of main pancreatic duct obstruction; (2) Santorini's duct promotes biliary pancreatic reflux during obstruction of the common channel and subsequent development of pancreatitis caused by infected choledochal secretions; (3) Santorini's duct may thus be both a protective morphological variant and a risk factor for pancreatitis dependent upon the site of stone impaction within the choledochoduodenal junction. PMID- 9222729 TI - Detection of antineutrophil cytoplasmic antibodies in primary sclerosing cholangitis: a comparison of the alkaline phosphatase and immunofluorescent techniques. AB - BACKGROUND: The reported prevalence of antineutrophil cytoplasmic antibodies (ANCA) in primary sclerosing cholangitis (PSC) varies considerably (26-85%). Part of this may reflect methodological differences but part may reflect the differences in the patient groups analysed. To resolve this issue we compared the sensitivity and specificity of the immunoalkaline phosphatase (IALP) and immunofluorescence (IF) techniques in four different populations. METHOD: Sera from four centres were tested blind on alcohol-fixed neutrophils using both techniques. PATIENTS: USA: 14 PSC, 14 primary biliary cirrhosis (PBC); Sweden: 32 PSC, 3 autoimmune hepatitis (AIH), 14 PBC, 11 chronic liver disease; Norway: 32 PSC, 14 AIH, 13 PBC, 1 hepatitis C. Italy: 8 PSC, 14 PBC, 8 viral hepatitis. Thirty-six normal healthy volunteers from Oxford, together with positive and negative controls, were also tested. RESULTS: The healthy controls were all ANCA negative. The diagnostic sensitivity and specificity, respectively, of ANCA for PSC using the IALP technique for the different test sera were: USA 71% and 93%, Sweden 66% and 96%, Norway 69% and 46%, Italy 50% and 95%. The diagnostic sensitivity and specificity, respectively, of the IF technique on the same sera were: USA 50% and 86%, Sweden 56% and 86%, Norway 47% and 61%, Italy 50% and 91%. Overall, combining all four groups, detection of ANCA using the IALP technique gave a diagnostic sensitivity of 66% with a specificity of 74% for PSC. In contrast, the IF technique gave an overall diagnostic of only 51% (P = 0.044, compared with IALP) with a specificity of 73%. Although overall the IALP technique was more sensitive than IF, the differences in sensitivity and specificity between the two techniques did not reach statistical significance for any individual group. Furthermore, the small differences in sensitivity between the four groups using either technique were not significant. However, the IALP technique had greater specificity in the US, Swedish and Italian groups compared with the Norwegian group (P < 0.05) whereas no statistically significant differences in specificity were noted between the groups using the IF technique. CONCLUSION: This study shows that the IALP method of ANCA detection is at least as sensitive as IF for the serological diagnosis of PSC. Indeed, combining data from all four centres, the IALP technique was significantly more sensitive than IF. We therefore recommend the use of the IALP technique, which is also easier to interpret and does not require the use of a specialist fluorescent microscope. The lack of a wide variation in sensitivity between IALP and IF for any individual patient group reported in this study suggests that the previously reported regional differences in ANCA prevalence in PSC of between 26% and 85% may be patient, related, rather than due to ethnic or methodological differences in ANCA detection, perhaps reflecting possible disease heterogeneity within PSC, or case selection bias. Further studies are needed to investigate this intriguing possibility. Such differences, if confirmed, will need to be taken into account when assessing the use of ANCA as a serological marker of PSC. PMID- 9222730 TI - Argon plasma coagulation (APC) in gastroenterology: experimental and clinical experiences. AB - BACKGROUND: Diathermy procedures are indispensable in interventional endoscopy. Argon beam coagulation is an innovative no-touch electrocoagulation technique in which high-frequency alternating current is delivered to the tissue through ionized argon gas. METHOD AND PATIENTS: Before clinical application, we conducted in-vitro studies to investigate the depth and diameters of tissue coagulation in fresh operative specimens from the stomach, small intestine and colon. Five different power/gas flow settings between 40 and 155 W and 2 and 7 l/min were used. The impact time (1-10s) and the incident angle of the probe (45 degrees and 90 degrees) were also varied. The maximum depth of necrosis was 2.4 mm, the maximum diameter 1.1 cm. No perforation occurred even in critical areas such as the colon and duodenum. We therefore performed argon beam coagulation in 66 consecutive patients. Two power/gas flow settings of 40 and 70 W and 2 and 3 l/min, respectively, were used. The impact time and incident angle were varied individually. RESULTS: In 49 of the 50 patients with oozing haemorrhage from angiodysplastic lesions, polypectomy sites, erosions or ulcers or oozing of blood due to vascular penetration by tumours, definitive haemostasis was achieved in one to two sessions. In all 11 patients with residual sessile adenoma tissue, complete removal was possible. Oesophageal patency was restored in all five patients with stenosing tumours. In one patient with angiodysplasia of the caecal pole, an asymptomatic accumulation of gas in the submucosa was observed which resolved spontaneously. In two patients with extensive oesophageal carcinoma, there was a transitory--also asymptomatic--accumulation of gas in the mediastinum and peritoneal cavity but no evidence of perforation. CONCLUSION: Argon plasma electrocoagulation is an effective and relatively low-cost alternative to laser therapy in gastrointestinal endoscopy. PMID- 9222731 TI - The age-related incidences of oesophageal carcinoma in intellectually disabled individuals in institutes in The Netherlands. AB - OBJECTIVE: An increased age-related incidence of oesophageal cancer in people with intellectual disability has been suggested by studies in the Netherlands. Gastro-oesophageal reflux disease (GORD), as documented by pH testing, occurs frequently in the intellectually disabled population, being found in nearly 50% of those with an IQ less than 50, while Barrett's oesophagus is found in about 15 26%. DESIGN: We compared the age-related incidence of oesophageal cancer in institutionalized, intellectually disabled individuals in the Netherlands with the age-related incidence in the general Dutch population. METHODS: Data were provided by the Netherlands Cancer Registry. The patient's institute physician was asked to complete a questionnaire about the diagnosis, which was endoscopically and histologically confirmed. RESULTS: The incidence of oesophageal carcinoma was 20 in 168,000 person-years. The expected incidence for oesophageal cancer, based on age-related incidence in the general population, was 7.0, resulting in a standardized morbidity ratio in the population with intellectual disability of 2.9 (confidence limits, 1.8-4.1; P < 0.001). Endoscopic findings were as follows: in 18/20 intellectually disabled carcinoma patients an adenocarcinoma was found; the remaining two patients had a squamous cell carcinoma. Barrett's epithelium was observed in nine patients (45%), eight (42%) of whom showed a peptic stricture as well. In 15 (75%) cancer patients reflux oesophagitis was found, accompanied in 14 cases by a hiatal hernia. CONCLUSION: A standardized morbidity ratio for oesophageal carcinoma of 2.9 was found in the intellectually disabled population as compared to the general population. Early detection and treatment of GORD in the population with intellectual disability is of paramount importance to prevent the development of Barrett's dysplasia and carcinoma. PMID- 9222733 TI - Morning and evening administration of pantoprazole: a study to compare the effect on 24-hour intragastric pH. AB - OBJECTIVE: To determine whether the time of administration (morning vs. evening) of pantoprazole influences the effect of a 40 mg dose upon intragastric pH in healthy subjects. DESIGN: Randomized, double-blind, two-period crossover study to compare intragastric pH following treatment with pantoprazole, 40 mg once daily for 7 days, the drug being given as either a morning or an evening dose before meals. METHODS: Intragastric pH was measured for 24 h on three occasions. The baseline recording was made 2 days prior to the first treatment period and subsequent measurements were made on days 6 to 7 of each period. Adverse events were recorded and fasting laboratory variables measured. RESULTS: Twelve subjects were evaluable for efficacy. Increases in median pH over 24 h were observed in all subjects with both dosage regimens. There was a greater increase from baseline in 24-h median pH values following morning than evening administration of pantoprazole (P < 0.05). This difference was due to a greater effect on median daytime pH (07.00-19.00 h, P < 0.01) compared with that after evening administration. No adverse events were reported and there were no clinically significant changes in laboratory variables. CONCLUSION: The study supports the recommendation of a once-daily morning dosage regimen of pantoprazole 40 mg in the treatment of acid-related diseases. PMID- 9222732 TI - Eradication of Helicobacter pylori in peptic ulcer disease with amoxycillin, 2.0 g, and omeprazole, 80 or 120 mg: a prospective randomized trial. AB - BACKGROUND: The appropriate dose of proton pump inhibitors needed for eradicating Helicobacter pylori by dual therapy is still controversial. DESIGN: The study was conducted as a single-blind, single-centre trial. METHODS: Fifty-four patients with active duodenal ulcers were treated with amoxycillin tablets, 750 mg three times daily, and omeprazole, either 40 mg twice daily (group 1) or 40 mg three times daily (group 2), for 14 days in a prospective randomized trial. H. pylori eradication was assessed 10 weeks after starting treatment. Biopsies were taken for rapid urease tests and histological analysis and 13C-urea breath tests were ordered. RESULTS: In both groups ulcer healing was complete in 96.3% of patients after 10 weeks. Ten weeks after starting treatment, Helicobacter pylori was eradicated in 76.9% of the patients in group 1 and 74.1% of those in group 2, as shown by rapid urease tests and histological analysis. In the subgroup of fully compliant patients (n = 49) the eradication rates were 80% and 79.2%, respectively. Hyperacidity significantly reduced the eradication rates. Patients showing successful H. pylori eradication were significantly older (59 +/- 14.0 years vs. 49 +/- 15.6 years; P = 0.025). Eradication rates were lower in smokers than in non-smokers (36.4% vs. 83.9%; P = 0.006). CONCLUSION: It is concluded that higher omeprazole doses should be reserved for younger patients and smokers; in others they are not needed. PMID- 9222734 TI - Medical prophylaxis of haemorrhage from oesophageal varices in patients with liver cirrhosis. AB - Haemorrhage from oesophageal varices is a life-threatening event in patients with liver cirrhosis. About 40-80% of patients surviving the first bleeding suffer a recurrence within 1 year. This high recurrence rate substantially contributes to the mortality in patients with liver cirrhosis. Therefore, various treatment regimens in both primary and secondary prophylaxis were studied. Most experience in medical primary prophylaxis was collected with beta-blockers, mainly propranolol. Treating patients with oesophageal varices with propranolol significantly reduces the incidence of first variceal bleeding. However, the effect on mortality is marginal, and primary prophylaxis is generally not recommended in these patients. Several studies support the hypothesis that medical prophylaxis with beta-blockers is more effective in reducing the rate of first oesophageal bleeding in patients with a high risk of haemorrhage, such as those with very large varices with red spots. A score to assess an individual patient's risk of variceal bleeding would be helpful, but until such a score has been validated, no general rule for this treatment decision can be given. In secondary prophylaxis, both beta-blockers and endoscopic therapy (sclerotherapy or ligation of the varices) are effective in lowering the rate of rebleeding. However, the effect on mortality was not significant in most studies. Several studies comparing the efficacy of medical prophylaxis and endoscopic treatment showed advantages of the endoscopic therapy with a greater reduction in recurrent bleeding episodes. However, medical prophylaxis with beta-blockers has the important advantage of being immediately effective, whereas endoscopic procedures provide the best protection against recurrent bleeding after complete obliteration of the varices. Therefore, in the first weeks and months of endoscopic therapy, additional treatment with beta-blockers may further reduce the risk of rebleeding. Only half of all studies on this topic reported a significant advantage with this combined therapy. Therefore, it seems reasonable to restrict this approach to patients with a high risk of rebleeding, such as patients with large sclerotherapy-derived oesophageal ulcers. PMID- 9222735 TI - Epidemiology and association with extra-gastrointestinal diseases. PMID- 9222736 TI - Extra-gastrointestinal diseases and Helicobacter pylori. PMID- 9222737 TI - Update on Helicobacter pylori research. Diagnosis. PMID- 9222738 TI - Uptake on Helicobacter pylori research. Pathogenesis and host response. PMID- 9222739 TI - Update on Helicobacter pylori research. Malignancies. PMID- 9222740 TI - Update on Helicobacter pylori research. Dyspepsia. PMID- 9222741 TI - Update on Helicobacter pylori research. Eradication. PMID- 9222742 TI - A caecal diaphragm in cystic fibrosis. AB - A right iliac fossa mass may be a difficult diagnostic problem in a patient with cystic fibrosis. We present a patient with such a mass who was thought to have a non-obstructing intussusception on clinical and radiological grounds. However, at laparotomy she was found to have a pathology not previously described in cystic fibrosis. She had a partial diaphragm almost blocking the lumen to her appendix. The differential diagnosis of a right iliac fossa mass is considered and the cause of her pathology discussed. PMID- 9222743 TI - Hepatocellular carcinoma in a non-cirrhotic coal miner with secondary haemochromatosis. AB - We report a case of hepatocarcinoma in a non-cirrhotic patient with secondary haemochromatosis. A 61-year-old man, who had been a coal miner for 36 years, presented with a well differentiated hepatocarcinoma without cirrhosis. The patient had a haemochromatosis with no family history of liver disease and a normal genetic study. Right hepatectomy was performed and 15 months later there has been no recurrence. This case confirms the possibility of hepatocarcinoma occurring in haemochromatosis patients without cirrhosis. The recognition of populations at risk of haemochromatosis is a necessity, and this should include coal miners. PMID- 9222744 TI - Food impaction in a duodenal diverticulum as an unusual cause of biliary obstruction: case reports and review of the literature. AB - We present two patients with upper abdominal complaints and symptoms of biliary obstruction. Endoscopic retrograde cholangiopancreatography showed that the common bile duct was obstructed by a juxtapapillary duodenal diverticulum filled with a food bezoar. There were no gallstones or other potential causes of obstruction. The bile flow was restored and symptoms disappeared after rinsing the diverticulum. Eventually, both of the patients were treated surgically because of recurrent symptoms. PMID- 9222745 TI - Choledochocele presenting with anaemia. AB - A 31-year-old man presented with abdominal pain and iron deficiency anaemia due to gastrointestinal blood loss. Endoscopic retrograde cholangiopancreatography (ERCP) revealed a choledochocele, located between the ampullary sphincter and the sphincters of the common bile duct and pancreatic duct. The choledochocele was removed surgically and appeared to be covered with duodenal mucosa. Gastrointestinal blood loss is explained by the extensive erosions found in the duodenal mucosa of the choledochocele. Choledochoceles should be treated by radical resection. PMID- 9222746 TI - Helicobacter pylori infection and upper gastrointestinal pathology in a British immigrant Indian community. PMID- 9222747 TI - Undernutrition and aging. AB - Diet restriction is a well-recognised method of slowing aging and prolonging life span in animals. However, previous studies of this have tended to start after weaning and the effects of prenatal or early postnatal diet restriction have rarely been considered. Here we summarise the existing literature, which suggests that reducing nutrition at this earlier stage of life has opposite effects, resulting in accelerated aging and a reduction in life span. These findings support emerging epidemiological evidence in man that poor nutrition in early life may programme accelerated aging and predispose to a variety of age-related diseases. PMID- 9222749 TI - A preliminary investigation of the effects of aging on the nerve cell number in the myenteric ganglia of the human colon. AB - We have examined the number of nerve cells present in the myenteric plexus of the human large intestine using a nonhistochemical method (Giemsa) in laminar preparations of the muscularis externa in two groups of subjects aged 20-35 and over 65 years. The collagen and elastic system related fibers in the myenteric ganglia were also qualitatively evaluated. The total number of neurons decreased in the old subjects by over 37%. The perikaryal area of most of the neurons in both the young and old subjects fell from 101 to 200 microns2. A ganglionic capsule was present and was thicker in the ganglia from the old subjects as were the septa within the ganglia. Both collagen and elastic system fibers were more numerous in the ganglia from the old subjects. We conclude that the decrease in neuron density with age is accompanied by an apparent increase in the fibrous components of the myenteric ganglia. PMID- 9222748 TI - No influence of aging on the circadian rhythm of erythropoietin in healthy subjects. AB - The diurnal rhythm in the circulating serum levels of erythropoietin (Epo) was investigated in a group of 20 clinically healthy subjects aged 30-55 years and in a group of 20 healthy subjects aged 55-75 years. Venous blood samples were drawn during the span of a whole day every 4 h, starting from midnight, for the determination of serum Epo levels by radioimmunoassay. Statistical analysis was carried out by means of the 'cosinor' method. Both groups presented a significant (p < 0.05) circadian rhythm in serum Epo levels, with maximum in the afternoon. The younger subjects had significantly (p < 0.05) higher mean daily levels and higher diurnal variations of serum Epo than older subjects; no difference (p > 0.05) was found between the groups regarding the peaks of the rhythms. These data confirm the presence of a circadian rhythm in serum EPO levels and suggest that the aging process does not influence the physiological diurnal fluctuations but modifies the mean daily levels and the amplitude of the diurnal variations, such as it occurs for many other variables. This behavior is an index of aging, but does not seem to have clinical implications. PMID- 9222750 TI - Determination of malonaldehyde in Alzheimer's disease: a comparative study of high-performance liquid chromatography and thiobarbituric acid test. AB - The concentration of malonaldehyde (MDA) was measured in human erythrocytes obtained from subjects suffering from senile dementia of the Alzheimer type (SDAT), non-demented elderly subjects, and from young controls by two methods: high-performance liquid chromatography (HPLC) and the thiobarbituric acid (TBA) test. The MDA concentration measured by HPLC showed significant differences between SDAT and young control groups (p < 0.01) and between SDAT and non demented elderly groups (p < 0.01), respectively. Nevertheless, significant differences were not exhibited between young control and non-demented elderly. Moreover, the rate of accumulation of TBA-reactive substances was not significantly different among the three groups. Our results indicate that the HPLC method is highly specific and accurate and distinguishes between true MDA and other aldehydes that may react with TBA. Significant increases in the concentration of MDA of SDAT subjects were found in comparison with the two other groups, indicating that the measurement of MDA in erythrocytes could be used as a marker of oxidative damage in Alzheimer's disease. PMID- 9222751 TI - Influence of visual control, conduction, and central integration on static and dynamic balance in healthy older adults. AB - Aging is associated with decreased balance abilities, resulting in an increased risk of fall. In order to appreciate the visual, somatosensory, and central signals involved in balance control, sophisticated methods of posturography assessment have been developed, using static and dynamic tests, eventually associated with electromyographic measurements. We applied such methods to a population of healthy older adults in order to appreciate the respective importance of each of these sensorial inputs in aging individuals. Posture control parameters were recorded on a force-measuring platform in 41 healthy young (age 28.5 +/- 5.9 years) and 50 older (age 69.8 +/- 5.9 years) adults, using a static test and two dynamic tests performed by all individuals first with eyes open, then with eyes closed. The distance covered by the center of foot pressure, sway area, and anteroposterior oscillations were significantly higher, with eyes open or closed, in older people than in young subjects. Significant differences were noted in dynamic tests with longer latency responses in the group of old people. Dynamic recordings in a sinusoidal test had a more regular pattern when performed eyes open in both groups and evidenced significantly greater instability in old people. These data suggest that vision remains important in maintaining postural control while conduction and central integration become less efficient with age. PMID- 9222753 TI - Deleterious network: a testable pathogenetic concept of Alzheimer's disease. AB - Cumulative evidence has indicated that a deleterious network is formed on the basis of close interactions among abnormal amyloid precursor protein (APP) metabolism, oxidative damage, compromised energy metabolism and impaired calcium homeostasis. A unifying hypothesis-the deleterious network hypothesis of Alzheimer's disease (AD)-proposes that the deleterious network, not any single factor, is the common pathway of AD. Aging and multiple genetic or environmental factors could trigger the network by promoting the occurrence of one or more of the key detrimental factors, leading to a number of pathological changes of the disorder. This new hypothesis appears to unify some major theories of AD, providing a sound basis for consistent explanations to a large variety of the observations about the disorder. In this article upto-date delineation of the novel theory is given. Three types of studies are also proposed for further determining the validity of the new hypothesis. Based on this theory, it is suggested that combinative applications of the approaches which can reduce the incidence of the four key pathological factors could become a new therapeutic strategy of AD. PMID- 9222752 TI - The age-related rise of plasma cholesterol in humans is not caused by a cell removal defect of LDL particles: 'in vitro' studies in peripheral mononuclear blood cells. AB - In healthy subjects with a normal body mass index, total plasma cholesterol, low density lipoprotein (LDL) cholesterol, and apoB lipoprotein levels are higher in older individuals (n = 34) than in younger subjects (n = 43). The two groups studied ranged in age from 60 to 93 years and from 17 to 30 years, respectively. The cholesterol synthesis rates of peripheral mononuclear blood cells from 14C acetate, total and unesterified cholesterol concentrations in freshly isolated cells, and the rates of uptake of pooled donor LDL, labeled with 125I- or 14C cholesteryl oleoyl ether, by cells derepressed in a lipoprotein-free medium were similar in both experimental groups. Thus, the rise of LDL cholesterol with age would seem to be likely secondary to the higher rate of very-low-density lipoprotein production, as shown by other investigators. PMID- 9222754 TI - Oligonucleotide micro-arrays for identification of unknown mutations: how far from reality? AB - The present review examines critically what has been published on oligonucleotide micro-arrays, from the point of view of detection of unknown mutations. We will first demonstrate the theoretical power of this new technique, and then argue that it is experimentally realistic. However, technical difficulties remain, and the proposed solutions for controlling the hybridisation specificity and for manufacturing the micro-arrays will be reviewed. Some are promising, but a complete integration of several of these solutions must be achieved before oligonucleotide micro-arrays become a universal tool for routine detection of unknown mutations. Nevertheless, sequence specific arrays should be available in the short term for the most frequently studied genes. PMID- 9222755 TI - Mutations in the GTP cyclohydrolase I and 6-pyruvoyl-tetrahydropterin synthase genes. AB - Tetrahydrobiopterin deficiencies are highly heterogeneous disorders, with more than 30 molecular lesions identified in the past 2 years in the GTP cyclohydrolase I and 6-pyruvoyl-tetrahydropterin synthase genes. The spectrum of mutations causing a reduction of these two biosynthetic enzymes is reviewed. Only three mutations, two present homozygously, are reported in the GTP cyclohydrolase I gene to cause the rare autosomal recessively inherited form of hyperphenylalaninemia. Most of the other mutations, which are scattered over the entire coding region for the six exon-containing GTP cyclohydrolase I gene, are observed in a heterozygous state with the wild-type allele and are associated with the dominant DOPA-responsive dystonia. Compound heterozygous or homozygous mutations spread over all six exons encoding the 6-pyruvoyl-tetrahydropterin synthase cause an autosomal recessively inherited variant of hyperphenylalaninemia, mostly accompanied by a deficiency of dopamine and serotonin. PMID- 9222756 TI - Multiple de novo MPZ (P0) point mutations in a sporadic Dejerine-Sottas case. AB - Dejerine-Sottas syndrome (DSS), a severe demyelinating peripheral neuropathy with onset in infancy, has been associated with mutations in either PMP22 or MPZ. Most cases of DSS are caused by a single heterozygous dominant point mutation. We identified three de novo point mutations in MPZ exon 3 in a sporadic DSS patient. These three point mutations occur on the same allele and result in three novel amino acid substitutions: Ile(85)Thr, Asn(87)His, and Asp(99)Asn. Our data raise the question as to the potential mechanism(s) involved in the formation of multiple point mutations at a given locus. PMID- 9222757 TI - Identification of mutations causing 6-pyruvoyl-tetrahydropterin synthase deficiency in four Italian families. AB - 6-Pyruvoyl-tetrahydrobiopterin synthase (PTPS) is involved in tetrahydrobiopterin (BH4) biosynthesis, the cofactor for various enzymes including the hepatic phenylalanine hydroxylase. Inherited PTPS deficiency leads to BH4 depletion, causes hyperphenylalaninemia, and requires cofactor replacement therapy for treatment. We previously isolated the human PTPS cDNA and recently characterized its corresponding gene, PTS. Here we developed PCR-based mutation analysis with newly designed primers to detect genomic alterations and describe five mutations, four of which are novel, in the PTS gene of four Italian families with affected individuals. The mutant alleles found included three missense mutations (T67M, K129E, D136V), a previously described triplet deletion (delta V57), and a single c-3-->g transversion in the 3'-acceptor splice site of intron 1, leading to cryptic splice site usage that resulted in a 12 bp deletion (mutant allele delta (K29-S32)). Except for K129E, all mutant alleles were inactive and/or unstable proteins, as shown by recombinant expression and Western blot analysis of patients' fibroblasts. The PTPS-deficient patient with the homozygous K129E allele had transient hyperphenylalaninemia, did not depend on BH4 replacement therapy, and showed normal PTPS immunoreactivity, but no enzyme activity in primary fibroblasts and red blood cells. In contrast to its inactivity in these cells, the K129E mutant was 2-3 fold more active than wild-type PTPS when transfected into COS-1 or the human hepatoma cell line Hep G2. K129E appears thus as a mutant PTPS whose activity depends on the cell type. PMID- 9222758 TI - Common founder mutation in the LDL receptor gene causing familial hypercholesterolaemia in the Icelandic population. AB - Haplotype analysis in 18 apparently unrelated families with familial hypercholesterolaemia (FH) in Iceland has identified at least five different chromosomes cosegregating with hypercholesterolaemia. The most common haplotype was identified in 11 of the 18 families, indicating a responsible for FH in the Icelandic population. By using single-strand conformation polymorphism (SSCP) and direct sequencing of amplified DNA, we identified a novel mutation (a T to a C) in the second nucleotide in the 5' part of intron 4 in the LDL receptor gene. This mutation was present in approximately 60% of the FH families (10/18), supporting the prediction of a common founder. These families could be traced to a common ancestor in half of the cases by going back no further than the eighteenth century. The mutation was predicted to affect correct splicing of exon 4, and analysis at the cellular level demonstrated an abnormal mRNA containing intron 4 sequence in lymphoblastoid cells from a patient carrying this mutation. Translation of the mRNA would lead to a premature stop codon and a truncated nonfunctional protein of 285 amino acids. The novel sequence change created a new restriction site for the restriction endonuclease NlaIII, and using this assay, 29 unrelated individuals with possible FH attending a lipid clinic for treatment were examined for this mutation. Two individuals in this group of patients were found to be carriers of this mutation, supporting the suggestion of a founder mutation. Using this assay for the detection of FH in the Icelandic population should identify > 60% of these individuals. PMID- 9222759 TI - The cystic fibrosis delta F508 gene mutation and cancer. AB - Following the observation that relatives of cystic fibrosis (CF) patients have an increased mortality due to leukaemia, a study was initiated to determine whether leukaemia patients had an increased prevalence of the delta F508 CF mutation. No increase in carriers were found among leukaemias; however the carrier frequency of the delta F508 mutation appeared to be reduced in patients with malignant melanoma analysed as a control group compared to the normal population. This paper extends our previous study and investigates several other common human tumours, including those of the colon, breast, and lymphoma tissue. Fewer than expected carriers remained among the melanoma group from South Wales. There were fewer than expected carriers among patients with colon cancer compared to the normal population. The prevalence of the delta F508 mutation was normal in lymphomas and leukaemias. PMID- 9222760 TI - Molecular heterogeneity of classical and Duarte galactosemia: mutation analysis by denaturing gradient gel electrophoresis. AB - Classical galactosemia is caused by one common missense mutation (Q188R) and by several rare mutations in the galactose-1-phosphate uridyltransferase (GALT) gene. The most common variant of GALT, the Duarte variant, occurs as two types, Duarte-1 (D-1) and Duarte-2 (D-2), both of which carry the sequence change N314D. D-1 increases, whereas D-2 decreases GALT activity. To study the molecular genetics of classical and Duarte galactosemia, we analyzed the GALT mutations in 30 families with classical galactosemia, in 10 families with the D-2 variant and in 3 individuals carrying the D-1 allele by denaturing gradient gel electrophoresis (DGGE). DGGE detected 59 of the 60 classical galactosemia alleles. Q188R accounted for 60%, K285N accounted for 28% of these alleles. Eight novel candidate galactosemia mutations were found. On all D-2 alleles N314D occurred in cis with two intronic sequence changes, on the D-1 alleles in cis with a neutral mutation in exon 7. We conclude that the mutations causing galactosemia are highly heterogeneous and that K285N is a second common galactosemia mutation in our population. PMID- 9222761 TI - An expanded histatin gene polymorphism and test of a possible disease resistant phenotype. AB - Histatins are small molecular weight salivary proteins that are important in the non-immune host defense system. Two frequent cis-linked coding-change mutations were previously described in exon 5 of the HIS2 gene of Blacks. The polymorphic mutant allele was termed HIS2(2) and the wild-type allele HIS2(1). We here describe two new non-coding change polymorphisms of the HIS2 gene: a deletion in intron 5 (7183-7198 del) and a C-->T mutation in exon 5 [C-->T (7104)] that characterize two new HIS2 alleles, HIS2(3) and HIS2(4) respectively. Both mutations occur on a HIS2(1) background. The HIS2(3) allele occurred only in Afro Americans, but not in 67 Japanese, 51 Chinese and 50 Whites. Among 66 random DNA samples from Afro-Americans, frequencies of HIS2(1), HIS2(2), HIS2(3) and HIS2(4) were 0.67, 0.22, 0.05 and 0.07 respectively, with a heterozygosity of 0.45. The frequencies of the HIS2(4) allele in 50 Whites and 50 Chinese were 0.06, and 0.1 respectively. In a comparison of 60 matched saliva and DNA samples from the Afro American population, the DNA-based mutation analysis reliably identified salivary histatin phenotypes. The salivary histatin polymorphism (inferred from PCR analysis) was used to test a biologically plausible hypothesis, that the mutant histatin phenotype (coded by the HIS2(2) allele) confers relative resistance to severe and fatal malaria. In a study of 185 Black Tanzanian subjects, there were no significant differences in HIS2(2) allelic frequencies between the various test groups: for 86 cerebral malaria subjects, 54 uncomplicated malaria subjects, and 45 combined asymptomatic parasitemia and health controls, HIS2(2) frequencies were 0.16, 0.17 and 0.17 respectively. Thus, there was no support for the hypothesis in this population. PMID- 9222762 TI - SSCP analysis: a blind sensitivity trial. AB - Studies of the sensitivity of SSCP analysis usually have been performed under conditions contrary to the rules of quality control trials and have produced widely different results. We have performed a blind trial of the sensitivity of SSCP analysis for the detection of mutations in fragments up to 500 bp in length under a fixed single set of electrophoretic conditions. The mutation detection rate was 84%. In addition, we have identified a second mutation in nine samples. All these mutations are polymorphisms, including a novel polymorphism 1248 + 52T/C first reported in the present work. PMID- 9222763 TI - Mucopolysaccharidosis type II: identification of six novel mutations in Italian patients. PMID- 9222764 TI - Mutations associated with hemophilia B in Turkish patients. PMID- 9222765 TI - Identification of a one-base germline deletion (codon 888 del C) and an intron splice acceptor site polymorphism in hMSH2. PMID- 9222766 TI - Homozygosity for the common Ashkenazi jewish Tay-Sachs +1 IVS-12 splice-junction mutation: first report. PMID- 9222767 TI - 24 bp deletion and Ala1278 to Val mutation of the ATP7B gene in a Sardinian family with Wilson disease. PMID- 9222768 TI - Novel mutation (A141D) in exon 4 of the CFTR gene identified in an Algerian patient. PMID- 9222769 TI - Projected cancer incidence rates in Bulgaria, 1968-2017. AB - BACKGROUND: Incidence predictions are applicable when planning preventive or screening health care programmes, diagnostic, treatment and rehabilitation facilities. The aim of this study was to predict future (1993-2017) incidence rates of the most common sites of cancer in Bulgaria: breast, cervix and corpus uteri in females, and lung, prostate and stomach in males. METHOD: Observed numbers of incident cases in the period 1968-1992 and observed (predicted) population size were employed. Age-cohort and age-cohort-period log-linear models were fitted to the observed data, assuming no change in the observed trends. RESULTS: The incidence rates for all the studied primary sites, except stomach cancer, were predicted to increase. The observed rates in the period 1988-1992 and the predicted rates in the period 2013-2017 per 100000 were in females: breast-from 38.8 to 64.6, cervix-from 12.8 to 19.3, corpus uteri-from 12.4 to 26.5. In males similar rates were: lung-from 41.0 to 73.8, prostate-from 10.1 to 15.0 and stomach-from 17.5 to 10.2. Due to the increasing incidence rates and ageing of the population the predicted number of new cases in the studied sites of cancer in the period 2013-2017 is 62% higher than that observed in the period 1988-1992. PMID- 9222770 TI - Hysterectomy and subsequent risk of cancer. AB - BACKGROUND: The objective of this retrospective cohort study was to assess the effect of hysterectomy on subsequent risk of cancer among 25,382 hysterectomized and a similar number of non-hysterectomized control women, registered in 1963 1976 in the Mass Screening Registry (MSR). METHODS: Cancer cases were obtained from the Finnish Cancer Registry (FCR) and standardized incidence ratio (SIR); the expected number of cases based on cancer incidence rates of the Finnish female population in 1967-1993, was used. Relative risk (RR) was calculated as SIR among the hysterectomized relative to non-hysterectomized women, adjusted for follow-up, education and parity. RESULTS: The RR estimates of non-genital cancers among women with any hysterectomy were approximately 5% higher than in the non hysterectomized cohort. Relative risks of rectal cancer (RR = 1.4, 95% confidence interval [CI]: 1.0-1.8) and thyroid cancer (RR = 2.1, 95% CI; 1.5-3.1) were significant and largest among women who had undergone total hysterectomy pre- or perimenopausally. Relative risk estimates of breast cancer were close to unity. CONCLUSIONS: Hysterectomy is not associated with any substantial protective or promoting effect on cancers in general. Elevated risk of papillary thyroid cancer following hysterectomy is biologically plausible, as there are reproductive and endocrinological causes of thyroid cancer. PMID- 9222771 TI - The effect of body mass index and oestrogen receptor level on survival of breast cancer patients. AB - BACKGROUND: Epidemiological studies have demonstrated that obesity and low oestrogen receptor level adversely affect survival from breast cancer. Few studies have examined the joint effects of these variables. METHODS: A cohort study was conducted in which 1169 breast cancer patients from the Northern Alberta Breast Cancer Registry were followed for an average of 4.4 years. A number of variables related to breast cancer incidence and prognosis were studied. Body mass index (BMI) was used as a proxy measure of obesity. RESULTS: A Cox regression analysis resulted in a final model with terms for size of tumour, number of positive axillary nodes, oestrogen receptor level, BMI, and age at diagnosis, plus an interaction term for node status and BMI. Having relatively less oestrogen receptor increased the hazard ratio by 1.8 (95% CI: 1.4-2.3); for woman with no positive nodes, being in the highest quartile of BMI increased the hazard ratio by 2.5 (95% CI: 1.2-5.2) compared to the lowest quartile. CONCLUSIONS: BMI and oestrogen receptor level independently influence survival from breast cancer, but BMI affects survival only in patients with no positive axillary nodes. PMID- 9222772 TI - Sexual risk factors for cervical cancer among rural Indian women: a case-control study. AB - BACKGROUND: The association between sexual behaviour and cervical cancer is well established. Despite a high incidence of cervical cancer in India, its role has not been widely investigated in Indian women among whom the rate of sexual promiscuity is known to be very low. A hospital-based case-control study was carried out to investigate the role of sexual risk factors in cervical cancer among rural Indian women. METHODS: A case-control design was used in which a total of 268 subjects, comprising 134 women with invasive cervical cancer as cases and 134 control women were studied. A multiple logistic regression model was used to analyse the data. RESULTS: The risk factors found to be associated with cervical cancer were early age at first coitus, extramarital sex partners of women and the time interval since first exposure. In a multiple logistic regression model, independent effects were observed for early age at first coitus, showing maximum risk in women who reported their first intercourse at < 12 years of age, compared to that of women at > or = 18 years (odds ratio [OR] = 3.5. 95% confidence interval [CI]: 1.1-10.9). Increased risk was also seen for women who had extramarital sex relationships (OR = 5.5, 95% CI: 1.5-19.5). The significant effect of early age at first coitus persisted after adjustment for latency period which also showed its independent risk association with cervical cancer in the multivariate analysis. CONCLUSION: These findings confirm the association between early age at first coitus and cervical cancer in women with a low rate of sexual promiscuity and define the role of these risk factors in cervical carcinogenesis among rural Indian women. PMID- 9222773 TI - Intrauterine device use and endometrial cancer risk. AB - BACKGROUND: Because intrauterine devices (IUD) invoke acute and chronic inflammatory responses in the endometrium, it is possible that prolonged insertion of an IUD could induce endometrial cancer. METHODS: We examined the relation between use of an IUD and endometrial cancer risk using data from a multicentre case-control study involving 405 endometrial cancer cases and 297 population controls. RESULTS: A total of 20 (4.9%) cases and 34 (11.4%) controls reported any use of an IUD. After adjustment for potential confounders, IUD use was not associated with an increased risk of endometrial cancer (RR = 0.56 for ever use; 95% CI: 0.3-1.0). Little reduction in risk was observed among women who last used an IUD within 10 years of the index date (RR = 0.84; 95% CI: 0.3-2.4) but risk was decreased among women who used an IUD in the more distant past (RR = 0.45; 95% CI: 0.2-1.0). Risk did not vary consistently with number of years of IUD use or with years since first use. Risk was not increased among women who used inert devices (RR = 0.46; 95% CI: 0.3-3.6) or those who used devices containing copper (RR = 1.08; 95% CI: 0.1-3.6). CONCLUSION: These data are reassuring in that they do not provide any evidence of an increased risk of endometrial cancer among women who have used IUD. PMID- 9222774 TI - Modelling of mortality data from a multi-centre study in Japan by means of Poisson regression with error in variables. AB - BACKGROUND: Death rates of particular categories in epidemiological studies are often based on a small number of occurrences which can be well described by a Poisson distribution. METHOD: We applied this model for the analysis of a multi centre study in five Japanese counties where the death rates of stomach cancer (ICD-9 code 151) in four age groups are known. In our example some covariates of the cases (e.g. plasma lycopene levels) are unknown values and are estimated from a randomly chosen collective. Therefore these values are subject to a sampling error. The inclusion of errors in variables (e-i-v) into the statistical model can adequately describe such a situation. The model is estimated in a Bayesian framework by means of resampling techniques. RESULTS: Based on the posterior distribution of the parameters the relative risk of stomach cancer is 0.46 (95% confidence interval: 0.23-0.79) comparing the maximum of the population medians of lycopene with the minimum. The estimated overdispersion is close to zero indicating only minor interference with other possible explanatory variables. In addition, we show that inclusion of e-i-v can give more accurate estimates of the parameters even from small sample sizes. CONCLUSIONS: Appropriate statistical methods allow the accurate estimation of relative risks from small sample sizes and from low number of cases. Lycopene plasma levels are good predictors for stomach cancer. PMID- 9222775 TI - Pre-existing ischaemic heart disease and ischaemic heart disease mortality in women compared with men. AB - BACKGROUND: In all 8353 women and 7058 men aged 45-64 took part in the Renfrew/Paisley survey in 1972-1976. They formed a prospective cohort study of a general population in the West of Scotland; an area with high ischaemic heart disease (IHD) mortality rates. The objective of this study was to investigate three indicators of pre-existing IHD and determine how they predicted subsequent IHD mortality in females compared with males. METHODS: Pre-existing IHD was ascertained by the Rose Angina questionnaire, a question on severe chest pain indicating evidence of previous IHD and an electrocardiogram at a screening examination. Mortality information for a 15-year follow-up period was available. RESULTS: Pre-existing IHD was higher at older ages and was less common in women than men. The risks of IHD mortality were doubled for those with a single cardiovascular indicator compared to those without, and were increased to fourfold for those with two or more indicators. Indicators of pre-existing IHD had high specificity and low sensitivity for subsequent IHD mortality in both women and men, and the positive predictive values for women in the oldest age group were similar to those for men in the youngest age group. CONCLUSIONS: Each indicator of pre-existing IHD was a useful predictor of subsequent IHD mortality in both women and men, even though IHD mortality rates were lower in women. The indicators obtained by questionnaire could be implemented in the primary health care setting to identify quickly those at risk who would benefit from further investigation and intervention. PMID- 9222776 TI - Strong mediators of social inequalities in risk of ischaemic heart disease: a six year follow-up in the Copenhagen Male Study. AB - OBJECTIVE: Large social inequalities exist in risk of ischaemic heart disease (IHD) in Western populations; inequalities which are only little accounted for by established risk factors. We wished to find out if some newly identified cardiovascular risk factors in concert with established factors might contribute further to the explanation. DESIGN AND SETTING: A 6-year follow-up in the Copenhagen Male Study. SUBJECTS: Some 2974 males aged 53-75 years (mean 63) without overt cardiovascular disease were included in the study. Potential confounders included were: alcohol, physical activity, smoking, serum lipids, serum cotinine, serum selenium, lifetime occupational exposure to soldering fumes and organic solvents, body mass index, blood pressure, hypertension, use of sugar in hot beverages, use of diuretics, and Lewis phenotypes. MAIN OUTCOME MEASURES: During the 6-year follow-up period (1985/1986-1991), 184 men (6.2%) had a first IHD event. Compared to higher social classes (classes I, II and III), lower classes (classes IV and V) had a significantly (P < 0.05) increased risk of IHD; age-adjusted relative risk (RR) with 95% confidence limits was 1.44 (1.1-1.9), P = 0.02. After multivariate adjustment for age, blood pressure, serum lipids, physical activity, and smoking, the RR dropped to 1.38 (1.0-1.9), P = 0.05. Some newly identified risk factors were significantly associated with increased risk of IHD as well as with low social class: a low serum selenium concentration, a low level of leisure time physical activity in midlife, long-term exposure to soldering fumes, and abstention from or a low consumption of wine and strong spirits. After adjustment for these factors also, the RR dropped to 1.12 (P = 0.54). CONCLUSIONS: The results of this study suggest that potentially modifiable risk factors associated with lifestyle and working environment are strong mediators of social inequalities in risk of ischaemic heart disease. PMID- 9222777 TI - Lifelong teetotallers, ex-drinkers and drinkers: mortality and the incidence of major coronary heart disease events in middle-aged British men. AB - BACKGROUND: To determine the risk of all cause mortality and the incidence of major coronary heart disease (CHD) events in lifelong teetotallers and in ex drinkers compared with occasional and regular drinkers. METHODS: A prospective study of middle-aged men drawn at random from one general practice in each of 24 British towns. Five years after the screening of 7735 men aged 40-59 years, 7167 provided further information on postal questionnaire enabling separation of non drinkers into lifelong teetotallers and ex-drinkers. RESULTS: During the follow up period of 9.8 years after the postal questionnaire there were 929 deaths from all causes and 490 major CHD events. Ex-drinkers exhibited increased cardiovascular and non-cardiovascular mortality; lifelong teetotallers showed the lowest cardiovascular mortality but a significantly increased non-cardiovascular mortality. After adjustment for confounding factors and pre-existing disease, the two non-drinking groups did not differ significantly in all cause mortality from occasional and regular drinkers (light, moderate or heavy) but lifelong teetotallers still showed a significant increase in non-cardiovascular mortality. Adjustment considerably attenuated the risk of both cardiovascular and non cardiovascular mortality in the ex-drinkers. In men without a diagnosis of CHD, lifelong teetotallers and ex-drinkers showed similar increased relative risk (RR) of heart attacks, with regular drinkers (combined) having a significantly decreased risk compared to occasional drinkers (RR = 0.78, 95% confidence interval [CI] : 0.64-0.96) and non-drinkers (RR = 0.69, 95% CI : 0.52-0.91). This decreased risk was small in absolute terms at around 2-3 major CHD events/1000 person-years. CONCLUSIONS: Lifelong teetotallers and ex-drinkers showed a significantly increased RR of major CHD events compared with regular drinkers, although this risk is small in absolute terms. Lifelong teetotallers have a low risk of overall cardiovascular mortality and an increased risk of non cardiovascular mortality. Non-drinkers constitute an unsuitable reference group in alcohol-related studies; occasional or even light drinkers may be more appropriate. Overall, there is no convincing evidence that light or moderate drinking has a protective effect on total or cardiovascular mortality in these middle-aged British men. PMID- 9222778 TI - Socioeconomic status within social class and mortality: a prospective study in middle-aged British men. AB - OBJECTIVE: It has been suggested that mortality differences between groups in society may be greater than are indicated by social class based on occupation. We have examined the relationship between social class and mortality using home and car ownership as additional indices of socioeconomic status within social class. DESIGN: A prospective study of a cohort of men representative of the social class distribution of middle-aged men in Great Britain. SETTING: One general practice in each of 24 towns in England, Wales and Scotland. SUBJECTS: Five years after the initial screening of 7735 men aged 40-59 years, 7262 men (94% of the original cohort) provided information on housing tenure and car ownership by completing a postal questionnaire. MAIN OUTCOME MEASURE: Deaths from all causes, cardiovascular, cancer and other non-cardiovascular causes during an average follow-up of 9.8 years (range 8.5-11.0 years) after the postal questionnaire. RESULTS: During the follow-up period there were 946 deaths from all causes among the 7262 men. The lowest mortality rates for all causes, cardiovascular, cancer and other non-cardiovascular causes were seen in non-manual social classes I and II. Manual social classes III and IV+V showed a significant 40% increase in risk of death compared to social classes I+II, even after adjustment for a wide range of risk factors (relative risk [RR] = 1.4, 95% confidence interval [CI]: 1.2-1.7 and RR = 1.4, 95% CI: 1.1-1.7 respectively). Within all social class groups, those owning both home and car showed lower rates than those who owned neither, even after adjustment for a wide range of risk factors and employment status. Compared with social classes I+II owning both home and car, all those not owning home and/or car, in each social group, showed a significant approximately twofold increase in risk of death. Adjusted RR for non-manual I+II = 2.1 (95% CI: 1.5 2.9), non-manual III RR = 2.0 (95% CI: 1.3-2.9), manual III RR = 1.8 (95% CI: 1.4 2.4) and manual IV+V RR = 1.8 (95% CI: 1.3-2.5). Similar relationships were seen in all major geographical regions of Great Britain. CONCLUSION: Mortality differences within society are greater than indicated by social class based on occupation alone. Irrespective of social class, men with greater material assets have lower rates of mortality from all causes than men less well endowed, independent of a wide range of lifestyle and biological factors. These findings suggest that mortality differences within our society are closely related to relative wealth. PMID- 9222779 TI - Is insulin an independent risk factor for hypertension? The Paris Prospective Study. AB - BACKGROUND: The link between hyperinsulinaemia and hypertension has been examined in few prospective studies and often diminished after adjustment for obesity, central adiposity and baseline blood pressures. METHODS: The incidence of hypertension was studied as a function of baseline insulin and glucose in 4149 Caucasian, non-hypertensive, non-diabetic middle-aged men from the Paris Prospective Study. Blood pressures were measured over the 3 years of follow-up; hypertension incidence was defined as systolic blood pressure > or = 160 mmHg or diastolic blood pressure > or = 95 mmHg or drug treatment for hypertension. RESULTS: Fasting and 2-hour glucose and insulin were predictive of hypertension, after controlling for the known risk factors: age, excessive alcohol consumption and family history of hypertension (FHH). However, after further controlling for body mass index and central adiposity (the iliac circumference), insulin was no longer predictive in men without an FHH. When weight variation was also taken into account, and further adjustment made for baseline blood pressure and heart rate, fasting insulin, only, was predictive when the subject had a weight increase, independently of FHH. Fasting glucose was predictive of hypertension except in the case of no change or weight decrease and a negative FHH; 2-hour glucose was predictive in the presence of a positive FHH. CONCLUSIONS: Insulin and glucose levels were both risk factors for hypertension, and this risk was enhanced in the case of a positive FHH. However, obesity, especially central obesity, confounded these relationships and might be an intermediary factor in the relationship between insulin and hypertension. PMID- 9222780 TI - Mortality as a function of temperature. A study in Valencia, Spain, 1991-1993. AB - BACKGROUND: Increased mortality is associated with both very low and very high ambient temperatures. This study assesses the relationship between daily numbers of deaths and variations in ambient temperature within the city of Valencia. METHODS: The daily number of deaths from all causes (total deaths and only those occurring in people aged over 70), as well as those deaths from specific causes (e.g. cardiovascular and respiratory diseases, malignant tumours and all causes except external ones) occurring within the city of Valencia were related to the average daily temperature using autoregressive Poisson regression controlling for seasonality, day of the week, holidays, air pollution, influenza incidence, and humidity. Temperature was measured within the regression model as two complementary variables: 'Heat' and 'Cold'; also taken into account were their delayed effects up to 2 weeks after measurement. RESULTS: Graphical analysis revealed a relationship between temperature and mortality according to the time of year. For the cooler months (November-April), the temperature at which mortality was lowest was the 'minimum' (i.e. around 15 degrees C), while for the warmer months (May-October), it occurred at around 24 degrees C. Because of this, a stratified analysis was undertaken with different values for the 'Heat' and 'Cold' variables according to which of the two seasons was involved. During the colder months of the year, higher temperatures tended to exert a rapid influence on mortality and the lower temperatures a more delayed relation. During the hot season it is the heat variable which more clearly manifests an effect, and this is prolonged over the two following weeks. Variations also occur according to age and cause of death. The effect of temperature is greater in persons aged over 70 years of age, and it is also greater in cases of circulatory and respiratory diseases. CONCLUSIONS: A statistically significant association has been found between temperature and mortality. This relationship is not monotonic, but mortality increases in proportion to the variance in ambient temperature from a range of temperatures that varies from winter to summer. PMID- 9222781 TI - The effects of own fetal growth on reported hypertension in parous women aged 33. AB - BACKGROUND: Data from the study of the British 1958 birth cohort, National Child Development Study (NCDS), has allowed wider investigation of the relationship between retarded fetal growth and risk of adult hypertension. METHODS: A history of self-reported hypertension was related to fetal growth in 3308 parous cohort members. Fetal growth, the measure used, is the difference in actual birthweight from that expected for the gestational age and subsequent adult height. The relationships were investigated both linearly and non-linearly adjusting for potential confounders. RESULTS: After adjustment for confounding factors, including adult weight for height, retarded fetal growth was associated with reported hypertension particularly when not confined to pregnancy. The latter was also associated with accelerated fetal growth, moderate or severe hypertension in the mother when pregnant with the cohort member, being relatively taller than your mother, and lack of educational qualifications. Hypertension confined to pregnancy was more likely among women who were themselves firstborn or older at childbirth. Neither maternal smoking during cohort's gestation nor cohort member's gestational age had a significant effect. The results are consistent with previous reports that fetal growth effects are less marked if gestation is short. CONCLUSIONS: The relationships between fetal growth and subsequent hypertension are extremely complex and variable, and need to be studied allowing for deviations from growth potential. Adult weight for height remains the strongest predictor of hypertension. The results suggest that losing weight is likely to have the same proportional benefit in women with and without a history of retarded fetal growth. PMID- 9222782 TI - Maternal smoking and urinary organ malformations. AB - BACKGROUND: In order to investigate whether an earlier reported association between maternal smoking during pregnancy and urinary organ malformations could be confirmed, a study was made using Swedish health registries. METHODS: Infants with kidney malformations (n = 483) and infants with other urinary organ malformations with no primary involvement of the kidney(s) (n = 719) were selected among 1117021 infants born 1983-1993 with known smoking exposure in early pregnancy. RESULTS: A moderate statistically significant association between maternal smoking and kidney malformations was found (adjusted odds ratio [OR] = 1.22, 95% confidence interval [CI] : 1.00-1.48). With the exception of genetic renal polycystic disease, the main subgroups of kidney malformations showed similar OR. For other urinary organ malformations, no association with smoking could be detected (OR = 0.93, 95% CI: 0.79-1.11). CONCLUSIONS: This study supports an association between maternal smoking during pregnancy and kidney malformations. No obvious confounders were detected but further work is needed before a causal inference can be made. PMID- 9222783 TI - The association of job strain and health behaviours in men and women. AB - BACKGROUND: This study examined whether self-reported job strain, defined by the Karasek model was associated with some early, potentially modifiable cardiovascular (CVD)-related health characteristics. METHODS: Data were gathered in a 1989 cross-sectional survey of 3843 randomly selected men and women employees of 32 worksites in Minnesota. Sex-stratified crude and multivariate analyses examined the independent association of job psychological demands, latitude, and the combination of these two job domains (i.e. strain), to body mass index (BMI), smoking, exercise, and dietary fat intake. RESULTS: Job demands was positively associated with smoking, smoking intensity, and high fat intake in men and with BMI and smoking intensity in women. Job latitude was positively associated with exercise in men and women. High-strain men smokers smoked more than other workers and high-strain women had higher BMI than other women. CONCLUSIONS: Overall, self-reported job demands, latitude, and job strain were associated with some CVD-related health characteristics, but the effects were not similar in magnitude or direction for all characteristics and they varied by sex. PMID- 9222784 TI - Alcoholism in social classes and occupations in Sweden. AB - BACKGROUND: A number of studies have shown variations in the occurrence of alcoholism between different socioeconomic groups and occupations, but it has not been clear to what extent this is related to the average alcohol consumption in the same socioeconomic groups or occupations. METHODS: The relationship between socioeconomic group and occupation and hospital discharge 1981-1983 due to 'diagnoses related to alcoholism' (AD) (alcohol psychosis, alcoholism, and alcohol intoxication) and liver cirrhosis was studied in a cohort of 375,035 men and 140,139 women in 13 counties in Sweden who had reported the same occupation in the censuses of 1960 and 1970. Data on alcohol consumption in different socioeconomic groups and occupations were collected from a conscription investigation and from the Swedish twin registry with data from 1969/70 and 1973 respectively. RESULTS: Intermediate or higher non-manual employees had lower risk of AD as well as of liver cirrhosis compared to manual workers for both sexes. Among males several, mostly blue-collar, occupations had increased relative risks of AD. A high level of association was found between the relative risks of AD and liver cirrhosis in socioeconomic groups, and the relative risk of AD in occupations, and the average alcohol consumption in the same socioeconomic groups/occupations among males. Such an association was not evident among women. CONCLUSION: The study shows, contrary to previous Swedish evidence, that there is a strong relationship between the incidence of alcoholism in socioeconomic groups and occupations and the average alcohol consumption in these groups among men. PMID- 9222785 TI - The interrelationship between income, health and employment status. AB - BACKGROUND: The aim of the study was to test the hypothesis that the relatively strong association between income and health compared to that between education/occupation and health, can partly be interpreted in terms of an association between employment status and health. METHODS: Health indicators used were the prevalence of one or more chronic conditions, and perceived general health. Data were generated from a postal survey, part of the baseline data collection of a Dutch prospective cohort study on socioeconomic inequalities in health. RESULTS: After controlling for differences in other socioeconomic indicators, the association between income and health was found to be stronger than that between occupation or education and health. Most of the difference in strength was found to be due to employment status, especially among men. Controlling for employment status, and controlling for the distribution of those with a long-term work disability in particular, reduced the risks of lower income groups, whereas the risks of lower educational and occupational groups hardly changed. CONCLUSIONS: These results suggest that the relatively strong association between income and health can for a large part be interpreted in terms of an interrelationship between employment status, income and health. More specifically, it is largely due to the concentration of the long-term disabled in lower income groups. This indicates the importance of the selection mechanism, as these groups are excluded from paid employment because of their health status, leading to a lowering of income. However, income was still found to be related to perceived general health after controlling for employment status especially among women. This suggests that an explanation in terms of an effect of material factors on health may also be important. PMID- 9222786 TI - Time to pregnancy and occupation in a group of Italian women. AB - BACKGROUND: It is estimated that 10-15% of all couples have experienced an infertility problem. The objective of this study was to evaluate the effect of occupation on the time interval between when a couple starts unprotected intercourse and a clinically recognizable pregnancy time to pregnancy (TTP). METHODS: Data from 622 women who successfully delivered in the week preceding the interview were analysed using the Cox proportional hazards regression. Thirty independent variables were included in the full model. RESULTS: Eleven per cent of women had to wait more than one year before conceiving (mean TTP = 6.7 months). The regression analysis showed that the most important determinants of TTP are the age of the woman (rate ratio = 0.44 for age 35+) and her parity (rate ratio = 1.39). TTP also increased significantly with maternal smoking (rate ratio = 0.77), and decreased with coital frequency (rate ratio = 1.24 for > or = 6 per month) and consumption of coffee (rate ratio = 1.29). None of the female occupational exposures has been found to have an independent statistically significant effect, while male occupation in industry and exposure to welding fumes were associated with an increase of TTP (rate ratio = 0.73 and 0.78, respectively). CONCLUSIONS: Female occupational exposures seem to have only a small effect on TTP compared with biological and lifestyle factors. The present data also suggest that work-related factors may have a bigger influence on male fecundity. PMID- 9222787 TI - Childhood asthma in four regions in Scandinavia: risk factors and avoidance effects. AB - BACKGROUND: The high and increasing prevalence of childhood asthma is a major public health issue. Various risk factors have been proposed in local studies with different designs. METHODS: We have made a questionnaire study of the prevalence of childhood asthma, potential risk factors and their relations in four regions in Scandinavia (Umea and Malmo in Sweden, Kuopio in eastern Finland and Oslo, Norway). One urban and one less urbanized area were selected in each region, and a study group of 15962 children aged 6-12 years was recruited. RESULTS: The prevalence of symptoms suggestive of asthma varied considerably between different areas (dry cough 8-19%, asthma attacks 4-8%, physician diagnosed asthma 4-9%), as did the potential risk factors. Urban residency was generally not a risk factor. However, dry cough was common in the most traffic polluted area. Exposure to some of the risk factors. such as smoking indoors and moisture stains or moulds at home during the first 2 years of life, resulted in an increased risk. However, current exposure was associated with odds ratios less than one. CONCLUSIONS: Our findings were probably due to a combination of early impact and later avoidance of these risk factors. The effects of some risk factors were found to differ significantly between regions. No overall pattern between air pollution and asthma was seen, but air pollution differed less than expected between the areas. PMID- 9222788 TI - Social environment, morbidity and use of health care among people with diabetes mellitus in Trinidad. AB - BACKGROUND: This study aimed to identify social characteristics associated with higher levels of morbidity from diabetes and their relationship to health care utilization. METHODS: During a 6-month period 1149/1447 (79%) subjects admitted to Port of Spain Hospital, Trinidad with diabetes responded to a structured interview. Data collection included social factors, diabetes-related morbidity and health care utilization. Analyses were adjusted for age, sex, ethnic group and self-reported diabetes duration. RESULTS: Of 12 indicators of morbidity, nine were more frequent in subjects with no schooling compared with those with secondary education. At ages 15-59 years, nine morbidity indicators were less frequent among subjects in full-time jobs compared with those not in employment. The association of educational attainment was explained by confounding with age, sex, ethnic group and diabetes duration but five morbidity indicators were associated with employment status after adjusting for confounding. The type of water supply in the home was generally not associated with morbidity. Each of the indicators of lower socioeconomic status was associated with less use of private doctors and with more use of government health centres. CONCLUSIONS: Morbidity from diabetes was greater in groups with lower socioeconomic status. While morbidity associated with lower educational attainment was mostly explained by older age; the results suggested the possibility that diabetes may contribute to unemployment of those in the labour force. Private care was less accessible to social groups with higher levels of morbidity and the availability of government funded health services was important for reducing inequalities in health care utilization. PMID- 9222789 TI - Do new cases of rheumatoid arthritis cluster in time or in space? AB - OBJECTIVES: To examine for evidence of clustering in time, in space and in space/time in the occurrence of rheumatoid arthritis (RA). SETTING: A population based incidence register of RA in the East Anglian region of the UK: population size 413,000. SUBJECTS: In all 687 new cases of inflammatory joint disease registered between 1 January 1990 and 31 December 1994 were studied. Population data were obtained from postcode areas by age and sex. ANALYSIS: Time trend analysis was conducted over the first 36 months and observed and expected distributions compared. Spatial clustering was based on comparison of observed distribution using map grid references to random expectation based on simulation. A similar procedure was undertaken for time/space clustering. RESULTS: There was no evidence of a time trend. There was only modest evidence of spatial clustering with non-random distribution observed in one area but there was no evidence of time/space clustering. CONCLUSION: Although a viral aetiology is the strongest candidate for RA, no evidence of a localized event in time was associated with disease development in this population. PMID- 9222790 TI - Retrospective assessment of occupational exposure to chemicals in community-based studies: validity and repeatability of industrial hygiene panel ratings. AB - BACKGROUND: Occupational hygiene panels are increasingly being used to rate retrospective occupational exposures to chemicals in community-based studies. This study aimed to assess the validity, reliability and feasibility of using such an expert panel in a brain tumour case-control study. METHODS: A panel of five experts was recruited to rate exposure to 21 chemicals for 298 job descriptions to investigate the level of agreement. Validity was assessed by comparing the ratings of the experts for 49 of the jobs with objective quantitative exposure data which existed for these jobs. Repeatability was assessed by comparing the results for 50 resubmissions. RESULTS: Specificity was high for reporting that exposure occurred (all above 90%), but sensitivity was variable with values between 48% and 79%. Weaker validity was found for rating exposure level and exposure frequency. The raters showed the greatest inter-rater agreement for exposure to three of the 21 chemicals considered (kappa = 0.64 for cutting fluids, kappa = 0.57 for welding fumes and kappa = 0.42 for lubricating oils). Intra-rater reliability, based on the 50 resubmitted jobs, was fair to good (kappa = 0.46, 0.73). CONCLUSIONS: The potential effect of exposure misclassification from using expert panels was quantified and found to be a significant source of bias. The optimum situation occurred where three of the five raters concurred, where an odds ratio of 2.2 was observed for a true odds ratio of 4.0. Future studies which plan to use expert panels should screen the experts for their suitability by validating their performance against jobs with known exposure data. PMID- 9222791 TI - Use of household pesticides and the risk of aplastic anaemia in Thailand. The Aplastic Anemia Study Group. AB - BACKGROUND: Aplastic anaemia is a severe blood dyscrasia that is more common in Thailand than in Western countries. Its a etiology remains poorly understood. METHODS: A case-control study was conducted in Bangkok and two rural regions of Thailand. The effect of household pesticides was evaluated among 253 incident cases of aplastic anaemia and 1174 hospital controls. RESULTS: A total of 54% of the cases and 61% of the controls were exposed 1-6 months previously. For most individual household pesticides and for groups classified according to chemical type (organophosphates, pyrethrins, and organochlorines), the relative risk (RR) estimates approximated 1.0; upper 95% confidence limits were below 2.0 for many comparisons. A significant association was observed for exposure to combination products containing dichlorvos and propoxur, with an overall RR estimate of 1.7 (95% confidence interval [CI]: 1.1-2.6); the estimate for regular use was 1.6 (95% CI: 0.9-2.9). CONCLUSIONS: The absence of a higher risk for the regular use of dichlorvos/propoxur reduces the credibility of the apparent association, which could well have been an artefact of multiple comparisons. We conclude that most household pesticides used in Thailand do not appear to increase the risk of aplastic anaemia. PMID- 9222792 TI - Evaluating the goodness of fit in models of sparse medical data: a simulation approach. AB - BACKGROUND: Epidemiological studies of rare events, which are common in the medical literature, often involve modeling sparse data sets. Assessing the fit of these models may be complicated by the large numbers of observed zeros in the data set. METHODS: Poisson models, fitted as generalized linear models, were used to investigate the referral patterns of patients suffering from end-stage renal failure in south west Wales. The usual method for assessing the goodness of fit is to compare the deviance with a chi 2 distribution with appropriate degrees of freedom. However, this test may be invalid when the data set is sparse, as the deviance values may be unusually low compared to the degrees of freedom. This would suggest that there is a problem with underdispersion when, in fact, the large numbers of zeros in the data set make the comparison with the chi 2 distribution unreliable. A simulation approach is advocated as an alternative method of assessing model fit in these situations. RESULTS: Three models are considered in detail here. The first modelled the total referrals in each of the 245 wards in the study area and included two explanatory variables. These observations were not unusually sparse and both the chi 2 goodness of fit test and the simulation methodology outlined here suggested that the model did not fit. The second model included the population 'at risk' as an offset and the model improved considerably. Both the chi 2 test and the simulation approach suggested that this model did fit. Finally, the data were disaggregated into five age groups providing 1225 observations and a very sparse data set. According to the chi 2 goodness of fit test, the deviance was very low suggesting that the model was underdispersed. Using simulated data, it was shown that the deviance was not unusually low and that the model fitted the data reasonably well. CONCLUSION: In cases where the data set being modelled is sparse, it is useful to test the goodness of fit of a Poisson model using a simulation approach, rather than relying on the chi 2 test. PMID- 9222793 TI - Modelling the impact of HIV disease on mortality in gay and bisexual men. AB - OBJECTIVE: To assess how HIV infection and AIDS (HIV/AIDS) impacts on mortality rates for gay and bisexual men. METHODS: Vital statistics data were obtained for a large Canadian urban centre from 1987 to 1992. Three scenarios were utilized with assumed proportions of gay and bisexual men of 3%, 6% and 9% among the male population age 20 years. For each scenario, non-HIV deaths were distributed according to the assumed proportion of the total population (3%, 6% or 9%) but 95% of HIV deaths were distributed to gay and bisexual men as this is the proportion of AIDS cases in gay and bisexual men in this centre. The main outcome measures of interest were age-specific patterns of death, life expectancy and life expectancy lost due to HIV/AIDS at exact age 20 years, and the probability of living from age 20 to 65 years. RESULTS: Estimates of the mid-period gay and bisexual population ranged from 5406 to 16,219 for the three scenarios, and total deaths in these men from 953 to 1703. Age-specific mortality was significantly higher for gay and bisexual men than all men aged 30-44. Life expectancy at age 20 for gay and bisexual men ranged from 34.0 years to 46.3 years for the 3% and 9% scenarios respectively. These were all lower than the 54.3 year life expectancy at age 20 for all men. The probability of living from age 20 to 65 years for gay and bisexual men ranged from 32% for the 3% scenario, to 59% for the 9% scenario. These figures were considerably lower than for all men where the probability of living from 20 to 65 was 78%. CONCLUSION: In a major Canadian centre, life expectancy at age 20 years for gay and bisexual men is 8 to 20 years less than for all men. If the same pattern of mortality were to continue, we estimate that nearly half of gay and bisexual men currently aged 20 years will not reach their 65th birthday. Under even the most liberal assumptions, gay and bisexual men in this urban centre are now experiencing a life expectancy similar to that experienced by all men in Canada in the year 1871. PMID- 9222795 TI - Indicators of fatal outcome in paediatric cerebral malaria: a study of 134 comatose Papua New Guinean children. AB - BACKGROUND: No comprehensive data on the clinical features and the prognosis of cerebral malaria in the South Pacific are available at present. We conducted a prospective study in children with cerebral malaria to assess the case fatality rate (CFR) in the region and to identify potential risk factors for death. METHODS: We recruited 134 children admitted to the Madang General Hospital between April 1991 and October 1993 with a strictly defined diagnosis of cerebral malaria. Besides clinical examination, we collected a blood sample for parasitological haematological and biochemical assessment. RESULTS: The CFR was 11.9% and the prevalence of residual neurological sequelae at discharge was 1.5%. The proportion of children presenting with deep coma (12%) or hypoglycaemia (17%) was lower in our study than in African ones, where severe complications are more frequent. Also mortality associated with hypoglycaemia on admission was lower. Clinical or laboratory conditions significantly associated with death were deep coma, malarial anaemia and hyperleucocytosis. CONCLUSIONS: All conditions associated with deep coma, such as shock, hypoglycaemia and acidosis, should be corrected. Also prompt administration of blood transfusions to patients with anaemia is likely to reduce the occurrence of death in Papua New Guinean children with cerebral malaria. PMID- 9222794 TI - The impact of immunization control activities on measles outbreaks in middle and low income countries. AB - BACKGROUND: The World Health Organization recommended strategy for responding to measles outbreaks in developing countries does not promote the use of immunization campaigns due to their high cost, disruptive nature and limited impact. Given the substantial morbidity and mortality associated with such outbreaks, a literature review was conducted as a basis for re-evaluating this policy. METHODS: Reports of supplementary immunization activities that were performed to control measles outbreaks in middle or low income countries were identified. The impact of the immunization activities on the course of each outbreak was evaluated by examining the data provided. RESULTS: Of 66 reports detailing a measles outbreak in a middle or low income country, 17 described supplementary immunization activities which included seven 'non-selective' immunization campaigns, three 'selective' campaigns and one use of an early 2 dose schedule. Eight of the reports commented on the impact of the response, five of which reported a reduction in outbreak morbidity. Only one of the reports, from an isolated island outbreak, provided sufficient data to support a possible reduction in outbreak-associated morbidity. CONCLUSIONS: There are limited data on the impact of measles outbreak immunization activities from developing countries. The available data do not support a change in the WHO recommended strategy for conducting a limited, if any, immunization response to such outbreaks. Immunization strategies which aim to prevent outbreaks may be more effective than campaigns to interrupt transmission of an outbreak which has already begun. PMID- 9222797 TI - Update of cancer surveillance of veterans in Massachusetts, USA. PMID- 9222796 TI - Prolonged breastfeeding and malnutrition. PMID- 9222798 TI - Childhood and adolescent leukaemia. PMID- 9222799 TI - Effectiveness of meningococcal vaccine in Brazil. PMID- 9222800 TI - Women with heart disease may need aggressive treatment for high cholesterol. PMID- 9222801 TI - Exercise lengthens life of postmenopausal women. PMID- 9222802 TI - Gun ownership associated with increased risk for murder and suicide. PMID- 9222803 TI - Gender-specific aspects of obesity. AB - Obesity, an increasingly prevalent and difficult-to-treat condition in the United States, affects more women than men. The distribution of body fat differs in the genders, with women carrying more fat "on" their frames and men more likely to exhibit central obesity, carrying weight "within" their frames. Changes in body distribution of fat occur with reproductive cycling and childbearing in women. Obesity in females can have important consequences for fertility, and menopause is accompanied by a significant increase in the waist-hip ratio in females, an important factor in raising their risk for coronary artery disease. Dieting and exercise have different consequences in the two genders: men, unlike women, maintain HDL levels and lose central obesity simply by dieting, while women require exercise in addition to restricted caloric intake to produce the same effects. Recent advances in elucidating the molecular pathobiology of obesity have not yet been examined with respect to gender. With very few exceptions, this is also true of studies examining the usefulness of anorexics in initiating and maintaining weight loss. More gender-specific information is required in these two last, newer areas of obesity-related investigation. PMID- 9222804 TI - High blood pressure in women. AB - There is a sexual dimorphism in blood pressure of humans and experimental animals: males tend to have higher blood pressure than females with functional ovaries, while ovariectomy or menopause tends to abolish the sexual dimorphism and cause females to develop a "male" pattern of blood pressure. Hypertensive male laboratory animals tend to have NaCl-sensitive blood pressure, while females are NaCl resistant unless their ovaries are removed, in which case NaCl sensitivity appears. The hormonal basis of NaCl sensitivity of blood pressure and of the sexual dimorphism of hypertension remains to be defined. Synthetic estrogens and progestins, as found in oral contraceptives, tend to elevate blood pressure, while naturally occurring estrogens lower it, or have no effect. Hypertension increases cardiovascular risk in women, as well as men, although the benefits of antihypertensive treatment have been more difficult to demonstrate in women. In the population of the United States, women are more aware of their hypertension, more likely to be treated medically, and more likely to have their blood pressure controlled. PMID- 9222805 TI - Antibiotic therapy in men with leukocytospermia. AB - OBJECTIVE: In this prospective study, we aimed to determine the efficacy of doxycycline and doxycycline plus ceftriaxone for the treatment of asymptomatic men with leukocytospermia. METHOD: Seventy men were included in this randomized and placebo-controlled study. White blood cell (WBC) concentrations were determined by peroxidase assay during the routine semen analysis. Twenty-four of 70 men with leukocytospermia were randomized as control group and administered placebo (group I), 25 received doxycyline alone (group II), and 21, doxycycline plus ceftriaxone (group III). Doxycycline, 100 mg, was given twice a day for ten days and ceftriaxone, 1 g, in two doses for only one day. After the treatment, semen analyses were repeated. RESULTS: After the treatment there was a significant decrease in WBC counts in groups II and III when compared with group I (P < .05). Both antibiotic regimens were found to be equally effective. However, the time needed for resolution of leukocytospermia (approximately 4 weeks) was similar between the control and treatment groups. CONCLUSION: Although both antibiotic regimens significantly, and equally, improved the white blood cell counts in men with leukocytospermia, they failed to treat the leukocytospermia, i.e., to bring the count below the limit of one million WBC/mL. Therefore, it is doubtful that antibiotic therapy should be recommended for asymptomatic men with leukocytospermia. PMID- 9222806 TI - Changes in immunologic variables (TNF-a, sCD8 and sCD4) during danazol treatment in patients with endometriosis. AB - DESIGN: The levels of TNF-a, sCD8, and sCD4 were measured in serum samples taken from 20 women. Blood samples for TNF-a, sCD8 and sCD4 levels were taken from 10 endometriotic women before treatment, during the last 15 days of a 6-month administration of danazol and 3 months after treatment. Blood samples were taken only once from 10 women without endometriosis (control group). RESULTS: TNF-a levels were higher in women with endometriosis before treatment compared to controls (P < .05). Administration of the drug significantly reduced the levels of TNF-a (P < .01) and sCD8 (P < .001). CONCLUSION: This study indicates that danazol significantly reduces endometriosis-associated autoimmune abnormalities. PMID- 9222807 TI - Effect of two anti-estrogens, clomiphene citrate and tamoxifen, on cervical mucus and sperm-cervical mucus interaction. AB - OBJECTIVE: To compare the effect of two ovulation-inducing agents, clomiphene citrate and tamoxifen, on cervical mucus and sperm-cervical mucus interaction. SUBJECTS AND SETTING: Forty couples with unexplained infertility attending infertility clinic. METHODS: Cervical mucus scoring and postcoital test done using the Moghissi system in a spontaneous cycle (control cycle) and with clomiphene citrate or tamoxifen (study cycles). RESULTS: Clomiphene citrate significantly decreased cervical mucus production, whereas tamoxifen significantly improved the total score. CONCLUSION: Tamoxifen is a better drug than clomiphene for ovulation induction in women with poor cervical mucus quality. PMID- 9222808 TI - Hardening of zona pellucida of mouse oocytes and embryos in vivo and in vitro. AB - OBJECTIVE: To evaluate the effect of in vitro culture on zona pellucida resistance in mouse oocytes and embryos. METHOD: Zona pellucida resistance was assessed by comparing duration of zona lysis in the presence of alpha chymotrypsin. The effects of artificial or physiological conditions of development were evaluated by comparing embryos in vitro with those left to reach the same stage of development in vivo. RESULTS: The time required for zona lysis of oocytes increased after 2, 24, and 48 hours in vitro (P < .001). The same observation holds true for oocytes left in vivo during 24 hours. Fertilization both in vivo and in vitro induced a major increase in zona resistance. At the two cell stage, in vitro culture did not harden the zona pellucida. At the morula stage and beyond, enzymatic lysis was slightly longer in vitro as compared to that of similar stages recovered from the genital tract. CONCLUSIONS: Our data indicate that in vitro culture conditions do not modify zona hardening in oocytes and only slightly increased zona resistance from the morula stage on. PMID- 9222809 TI - Mammalian cells adapted to growth at pH 6.7 have elevated HSP27 levels and are resistant to cisplatin. AB - HSP27 levels are elevated in two Chinese hamster cell lines and in a human melanoma cell line adapted to growth at pH 6.7. The level of HSP72 is elevated in the melanoma cell line but not in the hamster cell lines adapted to growth at pH 6.7. HSC73 levels are not elevated in any of the adapted cell lines. Low pH adapted cells from all cell lines are resistant to cisplatin. It is proposed that elevated HSP27 levels in low pH-adapted cells may play a role in resistance to hyperthermia and resistance to cisplatin. PMID- 9222810 TI - The use of esmolol in whole-body hyperthermia: cardiovascular effects. AB - Whole-body hyperthermia (WBH) is a well-described investigational adjunct to systemic chemotherapy for the treatment of advanced malignancies. The hemodynamic consequences of this physiologic state may include tachycardia, which can produce acute myocardial ischemia in patients with coronary artery disease. Ischemic heart disease is currently considered a contraindication to WBH. We chose to investigate the consequences of using a new beta 1-adrenergic antagonist, esmolol, to attempt to control the tachycardia associated with WBH. After institutional approval and patient consent, nine consecutive patients with normal cardiac function presenting for WBH with carboplatin infusion were studied. Along with standard monitors, radial arterial and oximetric thermodilution pulmonary artery catheters were placed. Patients were sedated and heated in a radiant warmer (Enthermics). Spontaneous ventilation was maintained and hemodynamic data were gathered at 37 degrees C, and at 41.8 degrees C (before, during and after esmolol infusion). Heart rate and cardiac output increased (by 46% (p = 0.001) and 35% (p = 0.04) respectively) while mean arterial pressure and systemic vascular resistance fell (by 18% (p = 0.02) and 44% (p = 0.006) respectively) during hyperthermia. Heart rate was significantly reduced during esmolol administration (mean dose 180 micrograms/kg/min) in the absence of changes in cardiac index and calculated oxygen delivery. Ventricular filling pressures and stroke work were unchanged. No heart failure, pulmonary edema, or other adverse event was observed. Hemodynamic changes seen during esmolol administration were completely reversed 15 min after the infusion was stopped. We conclude that the administration of moderate doses of esmolol is safe for this population of patients undergoing WBH, and that this technique raises the question of whether patients with ischemic heart disease could safely undergo WBH. PMID- 9222811 TI - Three-dimensional temperature control of palladium-nickel thermoseeds: a computer aided and experimental evaluation. AB - The capability of self-regulating thermoseeds to compensate for nonuniform cooling along their longitudinal axis has been investigated in this study. For this purpose a quasi three-dimensional computer model has been developed. Calculations of the temperature profile in tissue with nonuniform heat loss demonstrated a clear improvement in the longitudinal temperature control of PdNi seeds compared to constant power seeds. Further, two strategies for improved control of nonuniform cooling along the longitudinal axis of ferromagnetic seeds have been investigated: (1) application of a 'normal' undivided seed; and (2) division of a long seed in smaller segments of which each segment is able to respond more directly to local variations in the temperature distribution. Calculations with the quasi three-dimensional model showed that the loose segments do respond more directly to their close proximity. However, the equilibrium temperature of a segment in an area with high local blood flow will be relatively low due the limited heat production of PdNi thermoseeds. In the undivided seeds the high thermal conductivity of PdNi causes some levelling of the longitudinal temperature gradient in the seed. In addition, calorimetric experiments have shown that the heat production of a segmented seed is less effective because of a demagnetizing field. Also, the absence of PdNi between the segments reduces the heat production of the seed. PMID- 9222812 TI - Thermal effects of acrylic cementation at bone tumour sites. AB - The use of acrylic bone cement as an adjunct to surgical excision of giant cell tumour of bone appears to reduce the incidence of tumour recurrence. Possible mechanisms for this apparent tumour inhibition include cytotoxic effects from the methylmethacrylate monomer and tissue hyperthermia from the heat of polymerization of the cement. This work presents a method for the prediction of temperature fields and resulting tissue necrosis arising from the implantation of polymethylmethacrylate (PMMA) at the site of a curretted giant cell tumour of bone. This is accomplished using a two-dimensional model based on geometry obtained from digitized MRI images of the distal femur. A general-coordinate, non orthogonal grid generation technique is used and solutions are obtained with an alternating-direction implicit (ADI) finite-difference scheme. The nodal temperature histories are then used to evaluate the effect of variable defect size on the zone of thermally induced cell necrosis. The results suggest the depth of the necrotic region is quite sensitive to the size of the implant. In at least some cases, the heating effect is sufficient to cause significant necrosis of tumorigenic cells. Implanting a large mass of acrylic may risk overkill, damaging substantial amounts of healthy tissue. PMID- 9222813 TI - A local hyperthermia treatment which enhances antibody uptake in a glioma xenograft model does not affect tumour interstitial fluid pressure. AB - Solid tumours have an elevated interstitial fluid pressure (IFP) due to the lack of normal lymphatics, increased permeability of tumour vasculature and an expanding cell population within a potentially limited space. This elevated IFP has been proposed to be an important barrier to the delivery of drugs and marcromolecules. We have demonstrated that local hyperthermia (4 h, 41.8 degrees C) is capable of significantly enhancing the uptake of radiolabelled monoclonal antibodies (mAbs) in D-54 MG glioma xenografts grown subcutaneously in athymic mice. To determine if this increased uptake was attributable to alterations in the tumour IFP, pressure measurements using the wick-in-needle technique were made in tumours after hyperthermia treatment. These pressure measurements were taken at various time points from 4 to 90 h following the initiation of the hyperthermia and compared with pressures taken concurrently in untreated tumours. In addition, pressures were measured following a 2 h, 41.8 degrees C hyperthermia treatment, a protocol which does not result in elevated uptake of radiolabeled mAbs. No significant differences were seen at any time point in IFP measured in the tumours receiving either hyperthermia treatment when compared with untreated tumours. Thus, we conclude that the mechanism by which this hyperthermia regimen enhances mAb uptake in this human glioma xenograft model is not due to alternations in tumour IFP. PMID- 9222814 TI - Thermal enhancement of pirarubicin (THP-adriamycin) by mild hyperthermia in vitro. AB - It has been demonstrated that hyperthermia can enhance the cytotoxicity of several anticancer drugs. Pirarubicin (THP-adriamycin) is a less cardiotoxic derivative of adriamycin. The thermal enhancement of cytotoxicity of pirarubicin was studied at various elevated temperatures in vitro by using a Chinese hamster cell line, V79. Cell survival curves were obtained at elevated temperatures for V79 cells treated with heat given alone or in combination with pirarubicin, and D0, the treatment time to reduce cell survival from S to S/e, was obtained for each cell survival curve. The relationship between the logarithm of the D0 and the treatment temperature for cells treated with heat alone was biphasic with a breaking point at 43 degrees C, although that for cells treated with a combination of heat and pirarubicin was exponential with no breaking point. The slope of this relationship for heat alone > 43 degrees C was -0.72 +/- 0.094 h/degree C which was not significantly different from the slope for combined heat and pirarubicin, -0.64 +/- 0.032 h/degree C. The results indicated that the cytotoxicity of pirarubicin was thermally enhanced specifically by mild hyperthermia. Pirarubicin uptake into the V79 cells during hyperthermia was independent of the treatment temperature (37, 42, and 44 degrees C), suggesting that the thermal enhancement of pirarubicin was not due to the increased drug uptake at elevated temperatures. Based on these results, it is predictable that hyperthermia combined with pirarubicin is more effective below 43 degrees C which is easily achievable clinically. PMID- 9222815 TI - Bicarbonate-dependent proton extrusion in Chinese hamster ovary (CHO) cells adapted to growth at pH 6.7. AB - A CHO cell model is described for in vitro studies of the mechanisms underlying heat resistance in cells adapted to growth in acidic environments. Adaptation is defined as a loss of pH 6.7-induced sensitization to 42.0 degrees C cytotoxicity and it is accompanied with an elevation of steady-state intracellular pH (pHi). CHO cells cultured between 75 and 100 days at pH 6.7 became fully adapted (6.7G cells), and the adapted phenotype was maintained for at least 100 additional days of culture at pH 6.7. The surviving fraction (SF) of 6.7G cells heated (42.0 degrees C) at pH 6.7 was comparable with that of cells cultured at pH 7.3 (7.3G cells) and heated at pH 7.3, while the SF of 7.3G cells acutely acidified to pH 6.7 and heated was an order of magnitude less. Although this resistance of 6.7G cells to killing was observed at 42.0 degrees C, it was not observed at 43.0 and 45.0 degrees C. Both 6.7G and 7.3G cells were able to develop comparable levels of thermotolerance during 42.0 degrees C at their growth pHs. However, in agreement with the literature, development of thermotolerance was reduced in acutely acidified 7.3G cells. An acute acidification of only 0.2 pH unit from pH 6.7 to 6.5 also reduced the ability of 6.7G cells to develop thermotolerance during heating at 42.0 degrees C. The acquired 6.7G phenotype reverted to the 7.3G phenotype following 17 days of culture at pH 7.3. Amiloride (0.5 mM), an inhibitor of the Na+/H+ exchanger (NHE), did not sensitize 7.3G and 6.7G cells to 42.0 degrees at their growth pHs. However, sensitization was observed for acutely acidified 7.3G cells. This is consistent with the hypothesis that extracellular acute acidification causes a decrease in pHi, and that the recovery from that decrease is achieved in part by activation of the NHE. An elevation of steady state pHi, measured by analysing intracellular BCECF excitation spectra, was documented in a suspension assay for cells grown at pH 6.7 for 180 days. The elevation was bicarbonate-dependent (negligible in the absence of HCO3-, +0.17 pH units in the presence of HCO3-). These results suggest that the altered regulation of pHi in CHO cells adapted to pHe 6.7 is maintained by bicarbonate dependent processes. PMID- 9222816 TI - Murine modelling for cytokine induction. PMID- 9222817 TI - B lymphocyte development in the rabbit. PMID- 9222818 TI - Immunoglobulin gene diversification in cattle. AB - Research in several species has revealed that different types of mammals have evolved divergent molecular and cellular strategies for generating immunoglobulin diversity. Other chapters in this text have highlighted the specific characteristics unique to chicken, rabbit, mouse, human and sheep B lymphocyte development; namely indicating differences in the mechanisms of diversity and the site of primary B cell development. Studies of the bovine system have indicated that like the sheep system, the ileal Peyer's patch (IPP) is a likely chicken bursal equivalent, and is a site of primary B lymphocyte development. Substantial investigation in sheep has indicated that Ig diversity is created by untemplated somatic mutation and intense selective pressure (Reynaud et al., 1991). The frequency of alteration in the sheep Ig light chain gene locus also is characteristic of the bovine system, however, recent evidence from sequencing of bovine lambda light chain genes indicates that one mechanism that contributes to diversity is gene conversion, utilizing several pseudogenes located in the Ig locus (Parng et al., 1996). The mechanism by which this hyperalteration of Ig genes occurs in both sheep and cattle is poorly understood and is thus the focus of considerable investigation. The study of events in the IPP may also have informative ramifications for secondary diversification of the Ig repertoire by somatic hyperalteration in germinal centers. PMID- 9222819 TI - Avian B cell development. AB - Development of B cells in chickens proceeds via a series of discrete developmental stages that includes the maturation of committed B cell progenitors in the specialized microenvironment of the bursa of Fabricius. The bursa has been shown to be required for the amplification of the B cell pool and selects for cells with productive immunoglobulin rearrangement events. Other events regulating chicken B cell development such as lymphocyte trafficking and apoptosis are just beginning to be elucidated. Within the bursa, the variable regions of immunoglobulin genes of B cell progenitors are diversified by a process of intrachromosomal gene conversion, where blocks of sequence information are transferred from pseudo-V regions to the recombined variable regions of the immunoglobulin genes. Recently gene conversion has been determined to play a role in the diversification of the immune repertoire in other species. In this review we focus on the current understanding and recent advances of B cell development in the chicken. PMID- 9222820 TI - B cell development in mice. AB - The development and establishment of the B Cell Repertoire is the net result of both genetic and environmental forces. The primary event at the genetic level is Ig gene rearrangement resulting in numerous possible combination of genes which can be further modified by somatic events such as N segment addition and somatic mutation. Environmental forces in the form of self and exogenous Ags also shape the repertoire by positively or negatively selecting B cells according to the specificity of their Ig receptors. These are dynamic processes beginning with the earliest expression of immunoglobulins in fetal life and continuing throughout life. In this review we discuss the genetic and selective mechanisms responsible for differences in the early immune system compared to that of the adult. PMID- 9222821 TI - Human B cell biology. AB - Human B lymphocytes share one major distinctive feature with B cells of other higher animals, namely the ability to generate and secrete immunoglobulins. These highly specialized proteins are capable of tremendous diversity, and thereby account for much of our immune protection against invading organisms. Despite the great potential diversity possible in the specificities of immunoglobulin molecules, however, the binding of antibody to antigen initiates a limited spectrum of biologically important effector functions, such as complement activation and/or adherence of the immune complex to receptors on leukocytes. A variety of mechanisms have been elucidated that account for this, not all of which are shared by the different types of animals capable of making these proteins. The purpose of this chapter is to review the genetic, developmental, and physiologic mechanisms critical for human B cell expression of immunoglobulin. PMID- 9222822 TI - The genesis, tutelage and exodus of B cells in the ileal Peyer's patch of sheep. AB - The ileal Peyer's patch (PP) is a prominent lymphoid organ that extends 1-2 meters along the terminal small intestine of sheep. It is comprised of rapidly proliferating B cells that make major contributions to the animals total B cell system. The characteristics of this tissue in sheep have enabled a variety of novel approaches to studying both the B cell system and the contribution of PP to the mucosal immunity. The sheep ileal PP has characteristics that place it in a category similar to that of the thymus, bone marrow and the avian bursa of Fabricius. The ileal PP develops before birth and involutes while the sheep is still young. It produces B cells that populate the immune system but most of the large numbers of newly-formed B cells are rapidly destroyed by apoptosis. It has been concluded that this death is related to a selection event that examines each newly-formed cell. Antibody diversity in sheep is a post-rearrangement event, generated by the process of somatic hypermutation. A comparison of the lambda light chain gene from surviving and dying B cells indicates that when the PP is at its greatest size the dying cells have the characteristics of cells with high affinity receptors for a selecting ligand/antigen. It is proposed that if B cells proliferate in the continual presence of selecting ligand/antigen that the B cell receptor might develop a sufficient affinity to trigger apoptosis. This process might contribute to elimination of clones with high affinity for either self antigens or continually present environmental antigens. PMID- 9222824 TI - Comparative ruminal and total tract digestion of a finishing diet containing fresh vs air-dry steam-flaked corn. AB - Ten Holstein steers (465 +/- 6 kg) with cannulas in the rumen and proximal duodenum were used in a crossover design experiment to evaluate the influence of air-dry vs fresh steam-flaked corn on characteristics of ruminal and total tract digestion. The basal diet contained 77% steam-flaked corn (DM basis). Air-dry steam-flaked corn (SFC-AD) was obtained from a single batch that had been allowed to air-dry for 5 d before beginning the trial. Fresh steam-flaked corn (SFC-F) was produced daily Monday through Friday. Following production, the SFC-F was placed in air-tight polybags and stored at 4 degrees C until the time of feeding. There was little difference (P > .20) between SFC-AD and SFC-F with respect to site and extent of digestion of OM, starch, and fiber. Moreover, the two treatments did not differ (P > .20) in ruminal degradability of feed N. Apparent total tract N digestion was slightly greater (2.4%, P < .05) for SFC-F than for SFC-AD. Treatments did not affect ruminal pH (P > .20); however, VFA concentration of ruminal fluid tended to be greater (8.3%, P < .10) for SFC-F than for SFC-AD, indicating that the initial rate of fermentation may have been greater with SFC-F. Ruminal molar proportions of acetate were not affected by treatments (P > .20), but ruminal molar proportions of propionate tended to be greater (9.7%, P < .10) and molar proportions of butyrate tended to be less (10.0%, P < .10) for SFC-F than for SFC-AD. We conclude that the characteristics of digestion and the feeding value of steam-flaked corn is not altered by air drying before feeding. PMID- 9222823 TI - Effects of individual housing design and size on special-fed Holstein veal calf growth performance, hematology, and carcass characteristics. AB - The objective was to evaluate the effects of individual housing design (stalls vs pens) with widths of 56, 66, and 76 cm (2 x 3 factorial treatment arrangement) on growth, hematology, cleanliness, ambulation, abomasal hairball, and carcass measurements. Three groups of 36 Holstein bull calves (n = 108) were randomly allotted within group to treatments. There were no effects (P > .05) of housing design, width, or two-way interactions for BW, ADG, carcass weight, or dressing percentage. Blood samples were collected at approximately 33-d intervals. Mean values for hemoglobin, hematocrit, white blood cell count (WBC), and red blood cell count (RBC) were not different among treatments (P > .05), with the exception of d 28 hemoglobin, which was greater in the calves housed in 66-cm vs 76-cm stall. There were differences (P < .05) due to design and design x width effects for hind-quarter cleanliness; manure accumulation tended to be greater in pens vs stalls as width increased. There were increases (P < .05) in left front knee swelling scores as stall or pen size decreased; no important differences were observed in ambulatory ability among treatment groups. There were design effects (P < .05) for excitability scores, with calves in stalls being more excitable. There were no important treatment effects (P > .05) for liver, spleen, and lung condition, number of abomasal hairballs, or 0- and 24-h after slaughter flank or brisket color. These results indicate that housing designs and widths did not affect veal calf growth performance, WBC, RBC, hemoglobin, hematocrit, ambulation, or muscle color. PMID- 9222825 TI - Effect of forage:concentrate ratio on digestion and reproduction in primiparous beef heifers. AB - We evaluated the effects of high- (HF) and moderate- (MF) forage diets on digestive and reproductive characteristics in beef heifers. Thirty primiparous beef heifers were allotted by weight and backfat thickness to receive either 80:20 (HF) or 50: 50 (MF) forage:concentrate ratio diets from parturition to at least 90 d postpartum. Alfalfa hay and wheat straw were the forage sources and barley was the concentrate source. Equal daily amounts of ME were provided to all heifers by restricting intake of the MF diet. Digestibility of DM was greater (P < .001) for MF compared with HF diets, whereas NDF digestibility was not different. Dry matter and NDF digested daily was lower (P < .001) for MF than for HF diets. Ruminal fluid pH was lower (P < .05) for MF diets; however, the acetate:propionate ratio was not different. Serum insulin concentrations were greater for MF diets for all hours (P < .001) and weeks (P < .05) of sampling. Changes in weight, backfat thickness, and body condition score at 90 d postpartum were not different between treatments. Calf gain to 30 d, however, was greater (P < .10) for the MF than for the HF treatment (25.5 vs 20.7 kg). Maximum size of the ovulatory follicle was greater (P < .10) for cows receiving the HF diet than for cows receiving the MF diet. However, other aspects of ovarian follicular growth and wave dynamics and the intervals from parturition to first and second ovulation, first estrus, first service, and conception were not different between treatments. Shifts in energy supply from forage to concentrate had minimal effect on digestion and reproduction in first-calf beef heifers in this study. PMID- 9222826 TI - Influence of timing of gain on growth and reproductive performance of beef replacement heifers. AB - Our objective was to determine whether beef heifers could be developed by delaying the majority of weight gain until the last third of the developmental period before the onset of the breeding season. Spring-born Angus x Hereford heifers were used in each of two consecutive years and were allotted at weaning to gain either .45 kg/d for the entire developmental period (yr 1 = 159 d, n = 40; yr 2 = 168 d, n = 40; EVENGAIN) or to gain .11 kg/d from d 0 to 112, followed by .91 kg/d from d 112 to 159 (yr 1, n = 40) or d 168 (yr 2, n = 40; LATEGAIN). Body weights and condition scores were determined at d 0, 112, and 159 (yr 1) or d 0, 112, and 168 (yr 2). Heifers were subjected to a 60-d breeding season. Frame scores and pelvic areas were determined at the conclusion of the breeding season. Actual daily gains for EVENGAIN heifers for yr 1 and yr 2 were .60 and .51 kg/d, respectively. LATEGAIN heifers gained .25 and .05 kg/d during the restricted phases from d 0 to 112, followed by 1.14 and 1.32 kg/d during the accelerated growth phases for yr 1 and 2, respectively. Body weight at the onset of the breeding season and weight at puberty were not different between treatments in either year. Age at puberty did not differ in yr 1, but, age at puberty in yr 2 was delayed (P < .01) in LATEGAIN (406.9 d) compared to EVENGAIN (386.3 d) heifers. The LATEGAIN and EVENGAIN heifers had similar pelvic areas, frame scores, and body condition scores in each year. First-service conception rates of both groups were similar in yr 1 (55.5 vs 55.3%). In yr 2, LATEGAIN heifers tended (P = .18) to have an increase in first-service conception rate compared to EVENGAIN heifers (71.1 vs 56.4%). No treatment differences occurred in either average age of conception or overall pregnancy rates at the conclusion of the breeding season for either year. The LATEGAIN heifers were developed to a similar BW on 12 (P < .01) and 2.5% (not statistically significant) less feed for yr 1 and 2, respectively, compared to EVENGAIN heifers. We interpret these data to indicate that delaying the majority of weight gain until late in heifer development may decrease costs without detrimental effects on reproductive performance. PMID- 9222827 TI - Influence of dietary sulfur level on growth performance and digestive function in feedlot cattle. AB - Using ammonium sulfate, three levels of dietary S (.15, .20, and .25%, DM basis) were evaluated in a finishing trial with 108 yearling crossbred heifers (384 kg). The basal diet contained (DM basis) 4% alfalfa hay, 6% sudangrass hay, 74% steam flaked corn, 4% yellow grease, 6% cane molasses, and 6% protein-mineral supplement. Increasing dietary S decreased ADG (quadratic effect, P < .10), DMI (linear effect, P < .10), feed efficiency (quadratic effect, P < .10), diet NE (quadratic effect, P < .10), and longissimus muscle area (linear effect, P < .05). Six Holstein steers (218 kg) with cannulas in the rumen and proximal duodenum were used to evaluate treatment effects on characteristics of digestion. Treatment effects on ruminal and total tract digestion of OM and N were small (P > .10). However, ruminal digestion of ADF and starch was slightly lower (quadratic effect, P < .10), and postruminal digestion of ADF and starch was correspondingly greater (quadratic effect, P < .05) with supplemental S. Dietary S level did not influence (P > .10) ruminal synthesis of microbial N. Increasing dietary S did not influence (P > .10) ruminal pH or lactic acid. Increasing S decreased molar proportions of acetate (quadratic effect, P < .10) and increased molar proportions of propionate (linear effect, P < .10). We conclude that S in excess of .20% of dietary DM may have detrimental effects on growth performance and dietary NE. Excessive dietary S may also compromise carcass merit by decreasing longissimus muscle area. PMID- 9222828 TI - Genetic parameter estimates for growth and fleece characteristics in Targhee sheep. AB - Weaning weights at 60 (WW60) and 120 d (WW120), 60- to 120-d postweaning gains (PWG) for lambs weaned at 60 d, 120- to 365-d postweaning gains (YG) for lambs weaned at 120 d, fleece weights (FWT), and fiber diameters (FD) from 20 Targhee flocks were used to estimate parameters required for multiple-trait genetic evaluation. Flocks from western states (n = 10) recorded primarily WW60 (n = 1,762), WW120 (n = 5,961), YG (n = 2,388), FWT (n = 2,824), and FD (n = 2,000). Eastern flocks primarily recorded WW60 (n = 1,754) and PWG (n = 1,237). Heritability estimates were .01 for WW60 (.00 for western flocks and .07 for eastern flocks), .10 for WW120, .33 for PWG, .20 for YG, .41 for FWT, and .58 for FD. Additive maternal and maternal permanent environmental effects as a proportion of phenotypic variance were .10 and .09, respectively, for WW60 and .05 and .08 for WW120. In western flocks, maternal additive and permanent environmental effects on WW60 and WW120 were highly correlated (> .81), whereas WW120 and YG had a small positive additive genetic correlation (.19) but a negative residual correlation (-.34). Fleece weight had a genetic correlation of .50 with WW120 and YG. Supplemental analyses suggested that the observed genetic relationship between fleece weight and weaning weight arose primarily from a genetic association between additive direct genetic effects on fleece weight and additive maternal effects on weaning weight. Fiber diameter was nearly independent of body weights but had an undesirable additive correlation of .51 with FWT. In eastern flocks, WW60 and PWG had an additive correlation of .71 and a residual correlation of .15. PMID- 9222829 TI - Effect of total allelic relationship on accuracy of evaluation and response to selection. AB - Total allelic relationship (TA) as a possible alternative to the pedigree-derived additive genetic relationship (RA) is defined. The TA measures the actual allelic identity between individuals for loci segregating for the trait concerned. Its value was studied by simulation in populations of different family structure, different numbers of loci, different numbers of alleles per locus, and different heritability levels. The alternative types of relationship matrices were used in mixed model equations to derive best linear unbiased prediction estimates (EBV) of breeding values (BV). Accuracies of evaluations were calculated as correlations of EBV with true breeding values. In populations with random selection and mating, EBVTA derived using TA had higher accuracies than EBVRA derived using RA. In populations with selection, EBVTA was more accurate and resulted in higher responses than selection on EBVRA. We conclude that not accounting for variation in average measures of relationship and identity in state can be important sources of variance of prediction error, and taking account of them increases the accuracy of selection. PMID- 9222830 TI - Genotypic effects on norepinephrine-induced changes in thermogenesis, metabolic hormones, and metabolites in newborn calves. AB - Heat production was measured in newborn Angus-, Brahman-, and Tuli-sired calves born to Angus (n = 20) and Brahman (n = 26) dams, before (thermoneutral metabolic rate, TMR) and after norepinephrine (NE) infusion (peak metabolic rate, PMR), to assess genotypic effects on nonshivering thermogenesis in brown adipose tissue. Calves were fed pooled colostrum, fitted with jugular catheters, and placed in a temperature-controlled (37 degrees C) water immersion system. Heat production, determined by indirect calorimetry, and tympanic temperature were measured continuously in calves from approximately 3 to 6 h of age. Blood samples were collected at birth and at 0, 5, 20, 40, 60, 80, 100, and 120 min relative to NE infusion (35 micrograms.min-1.kg BW-1 for 4 min), and plasma was analyzed for metabolites (glucose, NEFA, and urea nitrogen [PUN]) and hormones (cortisol, triiodothyronine [T3] and thyroxine [T4]). Weight-specific TMR (cal.min-1.kg-1) was not affected by breed of sire or dam, although weight-specific PMR (cal.min 1.kg-1) was lower (P < .01) in Brahman-sired calves than in Angus- or Tuli-sired calves and was lower (P < .001) in calves born to Brahman rather than Angus dams. The reduction in weight-specific PMR due to the maternal Brahman influence was sire-breed dependent, and the reduction was largest (P < .01) for Tuli-sired (34.3%), intermediate (P < .05) for Brahman-sired (15.1%), and lowest (P > .25) for Angus-sired calves (4.1%). The PMR:TMR ratio was 1.80 and 2.21 +/- .06 in calves born to Brahman and Angus dams, respectively. Peak tympanic temperature was .6 degree C lower (P < .01) in calves born to Brahman rather than Angus dams. At birth, plasma NEFA concentrations were higher (P < .001) and glucose tended (P = .13) to be higher in calves born to Brahman rather than Angus dams. Cortisol, T3, and T4 concentrations at birth were higher in calves born to Brahman dams than in those born to Angus dams. These results suggest that calves born to Brahman dams may have less thermogenically active brown adipose tissue than calves born to Angus dams, which may contribute to the relative cold intolerance of calves with Bos indicus inheritance. PMID- 9222831 TI - Relationships between nutrition and foraging behavior of free-ranging sheep and goats. AB - Goats generally tolerate dry season forage conditions in northeastern Brazil's tropical deciduous woodland better than sheep do, but the mechanisms underlying sheep and goat response to wet and dry season conditions are not well understood. We evaluated aspects of nutrition and behavior of free-ranging sheep and goats in this woodland to improve our understanding of these mechanisms. Body weight changes, dietary botanical and chemical composition, and daily activities were measured for sheep and goats that foraged in this woodland during four defined periods of a year. Vegetation available to these animals was also measured. Although there was much less available vegetation during the wet season than during the dry season, body weights and dietary crude protein content of sheep and goats increased during the wet season and decreased during the dry season. During the four periods of the year, weight changes of sheep, and possibly those of goats, were positively related to the content of crude protein in their diets. Foraging times of sheep and goats were generally greater during the dry season than during the wet season and were positively related to available browse and inversely related to dietary crude protein content. Sheep and goats may have foraged longer during the dry periods because they were spending more time searching through the large quantity of low-quality vegetation for dietary items with relatively higher levels of crude protein. PMID- 9222832 TI - Effect of body composition at selection on reproductive development in large white gilts. AB - Fifty-four Large White gilts were used to determine the effect of body composition at selection (145 d of age) on the onset of puberty and subsequent reproductive development until 202 d of age. Gilts were assigned to one of three groups based on their backfat depth at selection: 10 to 12 mm (L), 13 to 15 mm (M), and 16 to 18 mm (F). All of the F gilts, 92% of the M gilts, and 67% of the L gilts reached puberty by slaughter at 202 d of age. Data from a subgroup (first 67% to reach puberty in each group; L = Lp, M = Mp, and F = Fp) was also used. The M (Mp) and F (Fp) gilts reached puberty at 172 d (166 d) and 170 d (166 d) of age, respectively, but the L (Lp) gilts at 184.5 d were 12 d (18 d) older than M (P < .05), Mp (P < .001), and F (P < .01), Fp (P < .001) gilts. The Lp (97.68 kg) and Mp (98.33 kg) gilts were lighter (P < .01) than Fp (108.72 kg) gilts at puberty. There were no differences (P < .05) among the L, M, and F gilts in terms of backfat depth or weight at puberty. The L (Lp) gilts had a mean of 1.16 (1.75) estrous cycles, which was lower (P < .01) than for M (Mp) and (P < .01) F (Fp) gilts, with 1.96 (2.29) and 2.25 (2.33) cycles, respectively. L (Lp) gilts had fewer (P < .05) follicles, 13.14 (12.63), than either M (Mp), 19.08 (18.71), or F (Fp), 18.25 (17.42) gilts. The number of corpora lutea was not influenced (P > .05) by grouping at selection, but Fp gilts had fewer (P < .05) corpora lutea than Mp or Fp gilts. Live weight at slaughter was not influenced (P > .10) by grouping at selection or subgrouping at puberty. The L gilts with a mean of 18.05 mm of backfat at slaughter were leaner (P < .05) than the F (21.66 mm) but not (P > .10) the M gilts (19.41 mm). Subgrouping had no effect. Fat deposition and protein deposition were higher (P < .05) in those animals that attained puberty. We conclude that the rate of fat and protein deposition seems to be one of the determinants of puberty attainment. PMID- 9222833 TI - Regulation of metabolism and growth during immune challenge: an overview of cytokine function. AB - Commercially reared food animals encounter serial pathogenic and nonpathogenic immune challenges throughout production. Because of the diversion of nutrients away from growth in support of immune-related processes, immune challenge is considered a major obstacle to animals' achieving their genetic potential for growth or efficiency of gain. Scientists now recognize that many metabolic processes respond directly or indirectly to proinflammatory cytokines. This cytokine-mediated "reprogramming" of metabolism is a homeorhetic mechanism that ensures an adequate supply of nutrients for proliferation of lymphocyte and macrophage populations, antibody production, and hepatic synthesis of acute phase proteins. Proinflammatory cytokines have been linked to altered nutrient uptake and utilization. Anabolic processes are interrupted, and companion catabolic activities are amplified. Furthermore, cytokines may influence prenatal growth and development, and to the extent that postnatal proliferation and differentiation of myogenic and adipogenic cells contribute to postnatal growth, cytokine regulation of these events may ultimately influence growth. The following discussion is an overview of the impact of immune challenge and proinflammatory cytokines on metabolism and growth. PMID- 9222834 TI - Effects of an endotoxin challenge on growth performance, carcass accretion rates, and serum hormone and metabolite concentrations in control pigs and those treated with recombinant porcine somatotropin. AB - Barrows were restrictively fed starting at 20 kg BW to determine the effects of endotoxin on growth performance of control and somatotropin-treated pigs. The following treatments were used: 1) daily i.m. vehicle injection until 55 kg BW; 2) daily i.m. injections of 100 micrograms of recombinant porcine somatotropin (pST)/kg BW, until 55 kg; 3) i.v. saline injections for 7 d consecutively starting at 60 kg BW; 4) i.v. injections of 1 microgram of bacterial lipopolysaccharide (LPS)/kg BW for 7 d starting at 60 kg BW; and 5) the combined LPS+pST treatment, with pST injections from 20 kg through the 7 d of LPS treatment. Pigs evaluated for LPS effects were fed to 60 kg anticipating a weight loss. Pigs were bled at 0800 and 1100 at 55 kg and on d 7 of LPS treatment. Rectal temperatures were taken on d 7. Treatment with pST increased ADG by 13 to 20% and improved feed:gain by 17 to 23% before LPS treatment. During the 7 d of LPS injections, ADG and feed:gain did not differ, although feed efficiency was impaired and variable. Rectal temperatures at 1100 were progressively increased: control < LPS < LPS-pST (P < .01). Protein accretion was improved 27% by pST treatment, and lipid accretion was decreased 45% before LPS. Lipid stores decreased (P < .01) after LPS treatment in the pST-treated pigs. Lipopolysaccharide treatment and(or) decreased feed intake reduced the hyperinsulinemia and hyperglycemia (P < .01) associated with pST treatment. These results indicate that LPS induced a simulated septicemia and that the effects were not negated by pST treatment. The observed hyperthermia was additive, possibly due to increased lean body mass induced by pST combined with the pyrogenic effect of LPS. PMID- 9222835 TI - An anti-adipocyte monoclonal antibody is cytotoxic to porcine preadipocytes in vitro and depresses the development of pig adipose tissue. AB - A mouse monoclonal antibody of the IgG2b subclass was raised against porcine adipocyte plasma membranes. This antibody did not cross-react in immunocytofluorescence with any tested cell-type or tissue other than porcine adipocytes. Complement-mediated cytotoxicity was demonstrated in primary cultures of porcine stromal-vascular cells. When the antibody and complement were added to already differentiated cultures, the treatment resulted in elimination of lipid filled preadipocytes, whereas an early treatment of cultures prevented the appearance of these cells. In vivo, injection of newborn pigs with 1 mg/kg of monoclonal antibody on d 2 and 5 of life produced a more than 20% reduction of subcutaneous and leaf fat lipids at 35 d of age, whereas the lipid content of the longissimus muscle remained unaffected. These results demonstrate that early systemic treatment of pigs with a specific anti-adipocyte antibody reduces the fat mass. In addition to their potential in vivo use, monoclonal antibodies directed against adipose determinants may be useful tools for studying adipocyte lineage. PMID- 9222836 TI - Effects of somatotropin and salbutamol in three genotypes of finishing barrows: growth, carcass, and calorimeter criteria. AB - We evaluated the combined use of 0 or 4 mg/d of recombinant porcine somatotropin (pST) and 0 or 2.75 ppm salbutamol in three genotypes of growing barrows (139 d old) differing in lean and lipid accretion potential. Treatments were in a 2 x 2 x 3 factorial arrangement, and the three genotypes tested were Meishan (M, n = 32, 49 kg), Duroc x White composite (D x Wc, n = 31, 62 kg), and Meishan x White composite (M x Wc, n = 31, 64 kg) pigs. Growth performance was evaluated over 28 d for individual pigs, and 20-h feed-deprived heat production was measured before slaughter (d 34). Daily pST injection increased ADG (+70 g/d) and reduced ADFI ( .61 kg/d) across genotypes (P < .05). Salbutamol increased (P < .05) ADG in M x Wc pigs (+146 g/d) but not in M pigs (-60 g/d) or D x Wc (+80 g/d) pigs. However, M pigs had the lowest ADG and ADFI, and M x Wc pigs not treated with salbutamol grew slower than D x Wc pigs (P < .05). Carcass protein and moisture accretion were additively (P < .05) increased by pST and salbutamol for D x Wc and M x Wc pigs. Meishan pigs had increased carcass protein and moisture accretion from pST, whereas only moisture accretion was increased by salbutamol (P < .05). The longissimus muscle area and semitendinosus weight increased as the percentage of M in the genotype decreased (P < .05), and both were increased by pST or salbutamol treatment (P < .001). Leaf fat was decreased more (P < .05) in M pigs than in D x Wc or M x Wc pigs with pST injection. The similar magnitude of leaf fat reduction between D x Wc and M x Wc pigs resulted in a mean genotype difference (P < .05), and salbutamol decreased leaf fat across genotypes. Oxygen consumption and heat production were increased by pST in M pigs more than in the crossbred genotypes, but CO2 production was reduced by similar magnitudes across genotypes, and salbutamol only tended to reduce CO2 production in D x Wc pigs. In general, these data indicate that pST and salbutamol result in additive increases in carcass lean composition; however, growth rate, carcass accretion, and various organ weights may vary among genotypes with salbutamol and pST treatment. PMID- 9222837 TI - Effects of somatotropin and salbutamol in three genotypes of finishing barrows: blood hormones and metabolites and muscle characteristics. AB - We evaluated the effects of recombinant porcine somatotropin (pST) and the beta adrenergic agonist salbutamol on plasam endocrine and metabolite profiles and muscle chemistry in three genotypes of growing barrows (n = 96, 139 d old). Treatments were in a 2 x 2 x 3 factorial arrangement, and main effects were pST (0 or 4 mg/d) and salbutamol (0 or 2.75 ppm); the three genotypes including purebred Meishan (M), 1/4 Duroc, 3/4 White composite (D x Wc), and 1/4 Meishan, 3/4 White composite (M x Wc). Individual pigs were injected daily with buffer or pST at 0700 and allowed ad libitum access to a corn-soybean meal diet (1.2% lysine) and water for 33 d. Plasma was obtained 4 h after injection and 3 h postprandially on d 0, 14, and 28 for determination of growth hormone (GH), insulin, IGF-I, glucose, urea N (PUN), NEFA, triglycerides, and cholesterol. Longissimus and semitendinosus samples were obtained, and protein, RNA, and DNA were quantified and loin chop shear force was measured. In general, plasma hormones and metabolites on d 14 and 28 were not affected by salbutamol in the absence of pST, although salbutamol tended to increase d 14 and 28 GH concentrations. Salbutamol lowered plasma IGF-I (d 14 and 28, P < .05), insulin (d 14, P < .01), and NEFA (d 28, P = .07) when pST was administered, although concentrations still exceeded those for control pigs. Salbutamol reduced (P < .05) IGF-I in M and M x Wc pigs, and GH was not changed in M pigs. Meishan pigs had a greater increase in glucose with pST than M x Wc or D x Wc pigs, although the effect was not consistent over time. Individual treatment with pST caused GH, insulin, IGF-I, glucose, NEFA, and triglycerides to be increased and PUN to be decreased on d 14 and 28. Cholesterol on d 14 and 28 was decreased by pST in M pigs, whereas no effects were found in the other genotypes. Muscle protein and RNA were increased by salbutamol and were consistently lowest for M pigs. Furthermore, pST did not affect muscle protein, but it increased RNA more in M pigs than in the others. Overall, pST and salbutamol seemed to act separately and by different mechanisms to alter muscle composition, but blood criteria generally representing fat metabolism (insulin, glucose, NEFA, triglycerides) were interactively affected. Meishan pigs tended to have greater changes in muscle and plasma composition with pST treatment than did M x Wc or D x Wc pigs. PMID- 9222838 TI - Genetic effects on beef tenderness in Bos indicus composite and Bos taurus cattle. AB - Bos indicus composite and Bos taurus cattle, originating from diverse production environments, were used to quantify genetic variation in marbling, 24-h calpastatin activity, and beef tenderness and to identify strategies for prevention of beef tenderness problems in Bos indicus composite cattle. Comparisons among 3/8 Bos indicus breeds (Braford, Red Brangus, Simbrah) revealed significant differences in marbling and 24-h calpastatin activity, but not in tenderness. Compared with Bos taurus cattle, 3/ 8 Bos indicus cattle had similar marbling scores but higher 24-h calpastatin activities. Also, beef from 3/8 Bos indicus composites aged more slowly from 1 to 7 d and was less tender at 4, 7, 14, 21, and 35 d postmortem than beef from Bos taurus cattle. However, beef from 3/8 Bos indicus cattle was relatively tender if it was aged for a sufficient period of time (21 d). The delayed response to aging and greater toughness of beef from 3/8 Bos indicus cattle was associated with Brahman breed effects and was not related to the Bos taurus germplasm source. Marbling was moderately heritable (.52 +/- .21) but exhibited positive genetic correlations with shear force at d 1 through 14 of aging, suggesting that, in these cattle, selection for increased marbling would have an unfavorable effect on beef tenderness. A low heritability estimate for 24-h calpastatin activity (.15 +/- .15), coupled with low genetic correlations between calpastatin activity and shear force at 7, 14, and 35 d, suggested that selection for low calpastatin activity would have little effect on aged beef tenderness. Panel tenderness and shear force at 7, 14, and 21 d were moderately heritable (.27 to .47), indicating that aged beef tenderness could be improved by direct selection (via progeny testing). Comparisons among Simbrah, Senegus x Simbrah, and Red Angus x Simmental steers showed that inclusion of a tropically adapted Bos taurus breed (Senepol) could be an effective strategy for preventing beef tenderness problems in heat-tolerant cattle. PMID- 9222839 TI - Predicting the yield and composition of mature cow carcasses. AB - Cow carcasses (n = 60) were selected based on conformation and external fat to develop more current and useful prediction equations for estimating yield and composition. Adjusted preliminary yield grade was highly correlated to percentage of the carcass as fat (.91), percentage fat in the total lean (.89), and percentage fat in the lean trimmings (.88) of carcasses from non-grain-fed mature cows. Equations for predicting percentage of the carcass as chemical fat had higher -R2 values than equations predicting other compositional end points. The "best" regression equation for predicting total yield (i.e., whole muscle cuts plus lean trimmings adjusted to 10% chemical fat) included hot carcass weight (HCWT), adjusted preliminary yield grade (APYG), longissimus area (LMA), and marbling (MARB), with R2 = .75 and residual standard deviation (RSD) = 2.47. A similar equation predicting total yield from unribbed carcass data included HCWT, APYG, and conformation (CONF) with R2 = .69 and RSD = 3.11. These two equations were applied to a test group of cow carcasses (n = 20), and the average difference between the actual and predicted total yield values from ribbed data and unribbed data was .45 and .83% of HCWT; simple correlations between the actual and predicted values were .74 and .69, respectively. These equations contain relatively simple independent variables to identify and more nearly represent current industry processing practices than equations previously available. PMID- 9222841 TI - Glucose tolerance, luteinizing hormone release, and reproductive performance of first-litter sows fed two levels of energy during gestation. AB - This experiment was conducted to determine the effects of gestational energy intake on glucose tolerance, LH profiles, and reproductive performance of first litter sows. Sixteen pregnant gilts were assigned to either high energy (11 Mcal of ME/d; H) or normal energy (6.5 Mcal of ME/d; N) diets during gestation. They did not receive the treatment diets until 35 d of gestation and had free access to the same commonly used diet during lactation. A glucose tolerance test (1 g glucose per kg BW) was conducted after 18 h of feed deprivation on 110 d of gestation and 15 d of lactation. Blood samples were collected at -10, -5, 0 (immediately before glucose infusion), 2, 4, 6, 8, 10, 15, 20, 25, 30, 40, 50, 60, 80, 100, 120, 150, and 180 min for assay of glucose and insulin concentrations. Blood samples were also taken on d 7, 14, and 21 of lactation and on 1 d postweaning for 6 h with 10-min intervals for LH characteristics. Sows in Group H had greater gain of BW and backfat during gestation (P < .001) and had less feed intake and greater weight loss during lactation (P < .002) than those in Group N. Sows in Group H (8.0 d) showed 1.6 d longer (P = .07) weaning-to estrus intervals than those in Group N (6.4 d). After glucose infusion, insulin levels in Group H were higher (P < .05) than those in Group N. Lower LH concentrations on 1 d postweaning were found (P < .05) in sows in Group H than in Group N. This study indicates that energy intake during gestation is negatively related to feed intake during lactation. Reduced feed intake during lactation of sows having high gestational energy impairs glucose tolerance, suggesting that interaction of insulin with LH secretion may influence weaning-to-estrus intervals. PMID- 9222840 TI - Effects of frame size and time-on-feed on carcass characteristics, sensory attributes, and fatty acid profiles of steers. AB - Weaned British x continental crossbred steers (n = 108, 7 to 8 mo of age) of medium or large frame were used in a replicated experiment with a 2 x 4 factorial arrangement of treatments over 2 yr. Management regimens consisted of backgrounding for 150 d and finishing for 0, 30, 60, and 90 d. Carcass data were collected, and samples from the longissimus muscle were analyzed for long-chain fatty acids, cholesterol, and the percentage of fat. Duration of finishing was a source of variation for hot carcass weight, marbling, percentage of kidney, pelvic, and heart fat, fat thickness, yield and quality grades, cooking loss, flavor intensity, and stearic, oleic, and linolenic acids (P < .05). Myristic, palmitic, and margaric acids were negatively correlated (P < .05) with juiciness and with "cowy" and "painty" taste characteristics. Frame score did not influence long-chain fatty acids; however, there was a relationship between long-chain fatty acids and management regimen. Results suggest that feeding steers a finishing diet up to 90 d after backgrounding for 150 d has a positive influence on carcass characteristics without affecting cholesterol. PMID- 9222842 TI - The effect of dietary lysine and valine fed during lactation on sow and litter performance. AB - Sows (98 first parity and 104 second parity) were used to determine the effects of dietary lysine and valine on lactation performance. Treatments were arranged in a 2 x 3 factorial with two levels of lysine (.80 or 1.20%) and three valine:lysine ratios (80, 100, or 120% of lysine). For all sows, increasing dietary lysine increased litter weaning weight (P < .001) and litter weight gain (P < .002) and reduced sow weight loss (P < .001). Litter weight gain tended (P = .22) to increase with increasing dietary valine, but the increase was not significant. Data were separated into two groups: sows that weaned 10 or more pigs and sows that weaned fewer than 10 pigs. For sows that weaned 10 or more pigs, litter weaning weight (P < .001) and litter weight gain (P < .001) increased and sow BW loss decreased (P < .001) when dietary lysine increased from .80 to 1.20%. For sows that weaned fewer than 10 pigs, increasing lysine had no effect (P < .77) on litter growth rate. For sows weaning 10 or more pigs, litter weaning weights (linear, P < .04; quadratic, P < .06) and litter weight gain increased (linear, P < .04; quadratic, P < .02) as dietary valine increased. For sows that weaned fewer than 10 pigs, maximum litter weight gain was observed at a valine:lysine ratio of 100% (quadratic, P < .13). These results demonstrate the need to increase dietary lysine and valine as litter weaning weights increase. High-producing sows that wean 10 or more pigs require increased dietary lysine and valine to maximize litter growth rate and minimize sow weight loss compared with sows weaning fewer than 10 pigs. The independent increases in litter weaning weights from adding lysine and valine suggest separate modes of action for these amino acids in high-producing sows. PMID- 9222843 TI - Effects of the interrelationship between zinc oxide and copper sulfate on growth performance of early-weaned pigs. AB - We conducted four experiments to examine the effects of adding zinc oxide (ZnO) and(or) copper sulfate (CuSO4) to diets for weanling pigs. In Exp. 1 and 2, weanling pigs (initially 5.3 kg and 19 +/- 2 d of age) were fed diets containing 250 ppm of added Cu (CuSO4) and either 110 or 3,110 ppm of added. Zn (ZnO). No differences (P > .10) were observed in either experiment for ADG, ADFI, or feed efficiency (G:F). In Exp. 3,240 pigs (initially 4.45 kg and 15 +/- 2 d of age) were used to determine the interactive effects of added dietary ZnO and(or) CuSO4. Dietary treatments were in a 2 x 2 factorial arrangement; Zn (165 or 3,000 ppm) and Cu (16.5 or 250 ppm) were the main effects. Pigs were fed a high nutrient dense diet from d 0 to 14 after weaning and a less complex diet from d 14 to 28 after weaning, both containing the same mineral fortifications. From d 0 to 14, pigs fed 3,000 ppm Zn, with or without 250 ppm Cu, had improved ADG (P < .01) compared with pigs fed the control (16.5 ppm Cu and 165 ppm Zn) or diets with only added Cu. From d 14 to 28, pigs fed the diet containing 3,000 ppm added Zn, without 250 ppm Cu, had greater ADG than pigs fed the other diets (Zn x Cu interaction, P < .01). In Exp. 4, 264 pigs (initially 4.17 kg and 12 +/- 3 d of age) were fed a high nutrient dense diet supplemented with 3,000 ppm of Zn (ZnO) from d 0 to 14 after weaning. On d 14, pigs were switched to the diets containing experimental mineral levels identical to those of Exp. 3. From d 14 to 28 after weaning, added Zn improved ADG but not when the diet contained 250 ppm Cu (Zn x Cu interaction, P < .05). Feeding 3,000 ppm of Zn from ZnO is a viable means of improving nursery pig performance, but additive responses to growth-promotant levels of CuSO4 (250 ppm Cu) were not observed. PMID- 9222844 TI - Failure of dietary bentonite clay, Silent Herder mineral supplement, or parenteral Banamine to alleviate locoweed toxicosis in rats. AB - To evaluate treatments purportedly beneficial for livestock grazing locoweeds (LW), growing rats were fed diets containing 10 or 20% whole-plant Oxytropis sericea (LW) with and without Silent Herder mineral mix (1.5% of diet) or bentonite clay (1.5% of diet). Pregnant female rats fed 10% LW were treated i.m. with Banamine (a prostaglandins suppressor) or saline. The LW contained swainsonine (430 micrograms/g DM) and elicited toxicosis within 10 d at intake of 2 mg/kg BW. In Trial 1, 96 immature male Sprague-Dawley rats (BW approximately 100 g) were fed commercial rat feed (CRF) with and without LW, as follows: 100% CRF, free choice; 100% CRF, restricted intake to equal average intake of rats consuming 10 and 20% LW; 90% CRF+10% LW free choice; and 80% CRF+20% LW free choice. Diets with LW contained either no supplement or supplemental mineral mixture (Silent Herder, 1.5% of diet) or added bentonite clay (1.5% of diet). Twelve rats received each of eight dietary regimens through 28 d. Locoweed depressed (P < .05) feed intake and BW gain, increased (P < .05) relative size of liver, kidneys, heart, spleen, and testes, and altered blood serum components (P < .05) indicating toxicosis. Dietary provision of Silent Herder or bentonite failed to benefit rats that ingested approximately 4 or 8 mg of swainsonine/kg BW daily through 28 d. In Trial 2, 68 young adult female Sprague-Dawley rats (approximately 230 g BW) were mated and directly assigned to three diets (100% CRF, free choice, 100% CRF, intake restricted slightly below average intake of diet by rats consuming LW, or 90% CRF+10% LW free choice) and two treatments (i.m. saline or i.m. Banamine at .25 mg/kg BW daily for 10 d) in a 3 x 2 factorial arrangement. Approximately half (31 of 68) of the impregnated rats were killed at d 10, when Banamine was discontinued, but diets were continued until the remaining females gave birth. Ingested LW provided approximately 2 mg swainsonine/kg BW daily and elicited toxicosis in 10 d, but LW failed to affect numbers of live concepti at d 10 (P > .5) or numbers of offspring at parturition (P > .10). Banamine did not alleviate LW toxicosis of dams (P > .10). Provision of Silent Herder or bentonite in the diet or Banamine i.m. had no benefit for rats fed toxic locoweed. PMID- 9222846 TI - Lack of a nocturnal rise in serum concentrations of melatonin as gilts attain puberty. AB - Twenty prepubertal crossbred gilts (Yorkshire x Hampshire x Duroc) weighing 98.1 +/- 4.2 kg at 5 mo of age were placed in an environmentally controlled room having a temperature of 18 degrees C and light:dark cycle of 12 h:12 h. Light intensity measured 700 lx at eye level to the gilts. Three mature ewes were penned adjacent to the gilts to serve as positive controls for the light-dark cycles. After a 30-d acclimation period, 10 gilts from the pool determined to be prepubertal (serum progesterone < 500 pg/mL) were fitted with surgically implanted jugular catheters. Blood samples were drawn at 1100 (4 h after onset of light), 1130, 1200, 2300 (4 h after onset of darkness), 2330, and 2400 for 4 d. On d 5 of sampling, gilts were transported in an open-bed truck for 15 min, returned to their original environment, and exposed to boars for 20 min. Boar exposure was repeated every day throughout the remainder of the experimental period. Blood samples were drawn from each gilt until 7 d after estrus or for 12 d in those gilts that did not exhibit estrus. Blood samples were drawn by venipuncture from the ewes during the entire experimental period. For each sampling day, within an individual gilt or ewe, means of serum concentrations of melatonin (MEL) for night (scotophase) and day (photophase) samples were calculated. After three replications were conducted, four classes of animals were obtained: ewes (n = 9); nonpubertal gilts (n = 10); and two classes of gilts that ultimately reached puberty (prepubertal [n = 16] and postpubertal [n = 16]). Across all gilts, only 65 of 406 bleeding periods (16.0%) had a nocturnal (scotophase) rise in serum MEL. The proportion of gilts expressing a nocturnal rise in serum MEL did not differ as gilts approached puberty (P > .05). Incidence of nocturnal rises of MEL was similar (P > .05) in gilts that attained puberty and gilts that did not attain puberty. Nocturnal rises in MEL were observed in 86.2% of the bleeding periods of ewes housed in the same environment. These data indicate clearly that nocturnal rises in serum MEL are not necessary for a gilt to attain puberty. PMID- 9222845 TI - The recruitment of ovarian follicles is enhanced by increased dietary intake in heifers. AB - The objective of this study was to determine whether dietary-induced changes in growth hormone (GH), insulin, and IGF-I alter the pattern of ovarian follicular development in heifers. Twenty-eight 2- to 3-yr-old Hereford-Friesian heifers were equally allocated (n = 7) to one of four dietary treatments: 1) Control (C) fed a maintenance diet as two meals per day; 2) twice maintenance (2M2) given as two meals per day; 3) twice maintenance (2M6) given as six meals per day; 4) feed deprived (F) fed maintenance requirements as for C, changed to straw ad libitum for 3 d from a synchronized estrus. On d 4 (estrus = d 0) heifers were transferred to grass silage (ad libitum access) until the end of the experiment. Blood samples were collected hourly for 10 h on d 1 and 3 and daily for an additional 14 d. Follicular development was monitored daily by ultrasonography until d 14. The number of small follicles (< 4 mm) was increased (P < .05) by 37% on d 1 and 2 in 2M2 and 2M6 heifers, with no carryover effect of nutrition to the second follicular wave. Number of medium-sized (4 to 8 mm) and large (> 8 mm) follicles did not vary (P > .05) among treatments. The FSH concentrations were not different (P > .05) among treatments. Insulin concentrations were higher (P < .05) in 2M2 and 2M6 heifers than in C or F heifers, with no carryover after the diet was changed. Increase in number of small follicles was independent of changes in FSH and IGF-I, negatively associated with GH, but positively associated with circulating insulin. These results indicate that a short-term increase in nutritional plane can affect follicular recruitment in cycling heifers. PMID- 9222847 TI - Influence of exogenous insulin before breeding on conception rate and litter size of sows. AB - We conducted two experiments on commercial farms to evaluate the effect of insulin administered to primiparous sows before postweaning estrus on subsequent reproductive performance. On the day after weaning in Exp. 1, 138 crossbred primiparous sows were assigned to receive saline or insulin (.4 IU/kg BW) once daily at the time of feeding for four consecutive days. Treatment did not affect interval from weaning to estrus (5.1 +/- .2 d) or percentage in estrus by 7 d after weaning (92.6%). Farrowing rate at second parity was increased by insulin (76.7 and 92.3% for saline and insulin treatments, respectively; P < .01). Litter sizes at second parity were not affected by insulin treatment. Experiment 2 was conducted on a different commercial farm using 491 primiparous crossbred sows. They were treated similarly to those in Exp. 1 with saline or insulin after weaning, except that treatment duration of 2 d of saline or insulin was included. Treatment did not affect the interval from weaning to estrus (overall, 9.2 +/- .5 d), percentage in estrus by 7 d after weaning (79.1%) or farrowing rate (90.2%). However, the number of total pigs born in the second litter for sows mated within 7 d after weaning was increased (P < .05) by one pig in sows treated with insulin for 4 d (10.3 +/- .3 pigs) compared with 2 d (9.1 +/- .3) and with saline (2 d and 4 d grouped together: 9.3 +/- .1). Third-parity litter size was not adversely affected by treatments applied before the second litter. In conclusion, manipulation with a metabolic hormone such as insulin may improve postweaning fertility by affecting aspects of ovarian follicle development or pregnancy establishment. PMID- 9222848 TI - Effects of suppressing cortisol following castration of bull calves on adrenocorticotropic hormone, in vitro interferon-gamma production, leukocytes, acute-phase proteins, growth, and feed intake. AB - The objective was to determine the effects of reducing the plasma cortisol rise in calves following castration on plasma ACTH concentrations, keyhole limpet hemocyanin (KLH)- and concanavalin A (Con A)-induced in vitro interferon (IFN) gamma production, white blood cell (WBC) numbers, neutrophil:lymphocyte (N:L) ratio, plasma haptoglobin and fibrinogen concentrations, ADG, and ADFI. Forty 5 mo-old Friesian bull calves (169 +/- 1.7 kg) were assigned to four treatments: 1) control (CON); 2) oral metyrapone administration (MET); 3) surgical castration at 0 h on d 0 (SURG); and 4) oral metyrapone administration and surgical castration (MET+SURG). Cortisol, ACTH, IFN-gamma production, haptoglobin, fibrinogen, ADFI, and ADG were not different between CON and MET animals. The MET+SURG calves had lower (P < .001) peak and mean cortisol during .25 to 1.5 h than SURG animals, but area under the cortisol vs time curve from 0 to 12 h did not differ (P > .39) between SURG and MET+SURG calves. Peak ACTH concentrations and area under the ACTH vs time curve from 0 to 6 h were greater (P < .05) for MET+SURG than for SURG calves. There were no differences between MET+SURG and SURG animals in IFN gamma production, WBC numbers, and ADFI. On d 1, MET+SURG and SURG animals had lower (P < .01) KLH- and Con A-induced IFN-gamma production and higher (P < .05) neutrophil numbers and N:L ratio compared with CON animals. Plasma haptoglobin on d 1 and 3 and fibrinogen concentrations on d 3 and 7 were elevated (P < .05) for MET+SURG and SURG compared with CON animals, whereas SURG animals had greater (P < .05) haptoglobin and fibrinogen concentrations than MET+SURG animals on d 7. The ADG of SURG calves was lower (P < .05) than that of MET+SURG calves during d 0 to 7. Metyrapone treatment partially suppressed cortisol and increased ACTH in castrated calves but did not alter the castration-induced suppression of IFN gamma and increases in neutrophil numbers and the N:L ratio. PMID- 9222849 TI - Circulating insulin-like growth factor I, insulin-like growth factor binding proteins, growth hormone, and resumption of estrus in postpartum cows subjected to dietary energy restriction. AB - The objective of this study was to determine whether serum concentrations of growth hormone (GH), IGF-I, IGF binding proteins (IGFBP), and glucose at wk 2 and 10 postpartum were associated with the ability of postpartum beef cows to resume cycling when maintained on a limited nutrient environment. Cows (n = 29) were individually fed either 130 or 170 kcal ME x BW-75 x d-1 during nonlactation and 170 or 210 kcal ME x BW-75 x d-1 during lactation for an average of 4.1 yr before sample collection. The proportion of cows that resumed estrus within 20 wk after parturition was less (P < .05) at the lower feeding rate (5 of 14) than at the higher feeding rate (11 of 15). Concentrations of IGF-I increased from wk 2 to 10 in cows that resumed cycling but not in cows that remained anestrous and were less (P < .05) at wk 2 and 10 in cows that remained anestrous compared to cows that resumed cycling. Circulating amounts of IGFBP-2 at wk 2 were greater (P < .05) and IGFBP-3 concentrations were lower (P < .05) in cows that remained anestrous compared to cows that resumed cycling. Cows on the lower feeding rate that did not cycle had lower body condition scores and greater concentrations of GH compared (P < .05) to other cows. At the higher feeding rate, body condition score and concentrations of GH did not differ between cows that did or did not resume cycling. Circulating concentrations of IGF-I and IGFBP-2 and -3 at wk 2 postpartum were indicators of the capacity of energy-restricted cattle to resume cycling after parturition. PMID- 9222850 TI - Effects of supplemental energy source and amount on forage intake and performance by steers and intake and diet digestibility by sheep. AB - Two levels of concentrate supplements containing different types of carbohydrates (corn-soybean meal, CSBM; wheat middlings, WM; and soybean hulls, SBH) were evaluated for effects on forage intake and performance in growing steers and total diet digestibility in sheep. In Exp. 1, 63 crossbred yearling cattle (298 and 377 kg initial BW for yr 1 and 2, respectively) were given ad libitum access to chopped bermudagrass (Cynodon dactylon [L.]) hay with no supplementation (CONTROL) or with 25 or 50% of projected total TDN intake from CSBM, WM, or SBH. In Exp. 2, digestibilities of organic matter (OMD) and neutral detergent fiber (NDFD) were determined with sheep fed levels of hay and concentrates similar to those used in the growth trials. Hay intake was 1.99% of BW for steers fed hay alone and averaged 1.93% of BW in steers fed supplements at the low level. At the high level of concentrate supplementation, hay intake was depressed (P < .001) to a similar extent (1.63% of BW) in steers supplemented with CSBM, WM, or SBH. AT the low concentrate level, shrunk ADG was similar (.63 kg/d) among supplements, but at the high concentrate level steers fed SBH had higher (P = .06) shrunk ADG (.95 kg/d) than steers fed CSBM (.76 kg/d). Body condition score (BCS) increased more (P = .06) for CSBM- and SBH- than for WM-supplemented steers. Total tract OMD was lower (P < .001) in sheep fed WM (54.8% for low and 56.9% for high supplementation levels) than in sheep fed CSBM (57.4 and 62.6%) or SBH (57.2 and 62.5%). Total tract NDFD was higher (P < .001) for the SBH (58.9% for low and 63.3% for high levels) diets than for CSBM (54.6 and 51.0%) or WM (54.6 and 51.8%) diets. Supplements containing highly digestible fiber (SBH) produced less negative associative effects than high-starch supplements (CSBM) when fed with bermudagrass hay at the high level (.8 to 1% of BW), but no differences were found at the low feeding level (.4 to .5% of BW). PMID- 9222851 TI - Performance of Angus and Brangus cow-calf pairs grazing Alicia bermudagrass and common bermudagrass-dallisgrass pastures. AB - This research was designed to examine genotype x environment interactions in cow calf growth performance of grazing animals. Angus and Brangus cow-calf pairs (minimum of six per breed) were allowed to rotationally graze (14-d intervals) treatment pastures from approximately May through early October in each of 2 yr. Treatment pastures contained relatively pure stands of Alicia bermudagrass (AP) or a mixed stand of common bermudagrass and dallisgrass (CDP). Forage allowance was equalized, using "put-and-take" cow-calf pairs, among forage and breed types at the initiation of each 14-d grazing interval. Forage samples were obtained in each paddock at the initiation of each grazing interval. Forage CP concentration was greater (P < .05; 13.5 vs 11.6%) and NDF concentration was less (P < .05; 63.8 vs 70.6%) for CDP than for AP. Daily weight loss was similar for Angus and Brangus cows, but it was greater (P < .05) for cows grazing AP than for cows grazing CDP. Calf ADG during the grazing season was 35% greater (P < .05) for CDP than for AP pastures and was 23% greater (P < .01) for Brangus than for Angus calves. Relative performance of Angus and Brangus cow-calf pairs was consistent between forages; no breed x forage interactions were observed. PMID- 9222852 TI - Analysis of lupin seed protein digestibility using gel electrophoresis and immunoblots. AB - Proteins from the seeds of 12 cultivars of three lupin species were analyzed by gel electrophoresis. Similarities between cultivars of the same species were noted. Antibodies raised against the three major globular proteins, conglutin alpha, beta, and gamma, of Lupinus albus cv. Ultra were used to probe immunoblots of crude extracts. The immunoblots revealed variations between cultivars not previously resolved and identified which protein-subunits were derived from which conglutin. In vitro digestibility studies were done on four of the lupin cultivars. During the digestion of these cultivars, the large protein units were shown to be degraded to smaller intermediates with specific molecular sizes. Some of the intermediate protein subunits were identified as being derived from conglutin beta. The digestibility of the four cultivars, based on the amount of identifiable protein in the ruminal fluid digest at 9 and 24 h, showed Ultra > Primorski > Juno > Danja. From this study a novel system of analyzing protein digestibility was devised. PMID- 9222853 TI - The amino acid composition of protein feedstuffs before and after ruminal incubation and after subsequent passage through the intestines of dairy cows. AB - The amino (AA) profile of eight feedstuffs (two samples of fishmeal, soybean meal [SBM], formaldehyde-treated SBM, sopralin, cotton-seed meal [CSM], rapeseed meal [RSM], corn distillers grains [CDG], and corn gluten feed [CGF]) was determined before and after ruminal incubation for 8 and 12 h in three lactating Friesian cows using nylon bags. The profile of AA disappearing during intestinal passage was also measured by inserting the bags into the duodenum through T-piece cannulas after ruminal incubation and recovering them in the feces. The AA profile changed for all feedstuffs, except sopralin, following ruminal incubation. Changes were greater for the more degradable feedstuffs. The profile of AA disappearing during intestinal passage was generally similar to the profile after ruminal incubation, but some differences were found with feedstuffs that had low (< 84%) intestinal disappearance of AA (RSM, CDG, CGF). For the other feedstuffs, intestinal disappearance of AA was greater than 93%. The two fishmeal samples had the highest concentrations of methionine and lysine in their residues after ruminal incubation, whereas CDG and CGF had low lysine concentrations. Residues of these latter two and RSM were quite high in methionine concentration, whereas residues of SBM, sopralin, and CSM had the lowest concentrations. Corn distillers grains had 13% of its AA remaining after ruminal incubation followed by intestinal passage. These results show that feedstuffs vary in the proportion of their AA that escape ruminal degradation, in the profile of those AA, and in their intestinal digestibility. These factors should be considered in protein evaluation systems and in assessment of the protein quality of feedstuffs. PMID- 9222854 TI - Oxygen consumption by and blood flow across the portal-drained viscera and liver of pregnant ewes. AB - The energy requirement of ewes increases during pregnancy. In late pregnancy, approximately 40% of the increase in heat production can be attributed to increases in heat production by nonreproductive tissues. The objective of this study was to determine the pattern of oxygen consumption by the portal-drained viscera (PDV) and liver during pregnancy to allow for an estimation of the extent to which these tissues contribute to the increase in energy requirement. Nineteen multiparous ewes were individually penned and allowed ad libitum access to an alfalfa hay-based diet. Catheters were surgically placed in the portal vein, a branch of the hepatic vein, a mesenteric vein, and the abdominal aorta. Oxygen consumption by the PDV and liver were subsequently measured before breeding and at 6, 19, 39, 61, 82, and, 103 d before lambing. Hepatic arterial blood flow was not influenced by litter size (P = .89) or stage of pregnancy (P = .28). Portal and hepatic venous blood flow peaked 19 d before lambing. Oxygen consumption by the PDV and liver increased with increased ad libitum feed intake. The increase in hepatic oxygen consumption occurred approximately 63 d earlier in ewes with twins than in ewes with a single fetus independent of changes in feed intake. Hepatic oxygen consumption increased with duration of gestation and was estimated to account for 40% of the heat production not associated with the gravid uterus. PMID- 9222855 TI - Effect of shipping and chromium supplementation on performance, immune response, and disease resistance of steers. AB - Forty-eight Angus crossbred steers (263 +/- 2 kg initial BW) were blocked by weight and randomly assigned within weight group to treatment. Treatments consisted of control or .4 mg of supplemental Cr as Cr-nicotinic acid complex/kg of DM. Steers were fed diets containing 90% corn silage (DM basis) and 10% of a soybean meal-mineral-vitamin supplement. After 56 d on the dietary treatment, half of the steers in each treatment were transported 343 km and unloaded in an unfamiliar location. The next day, d 58, shipped steers were returned to the feedlot (50 km). On d 58 after shipped steers were returned to the feedlot, all steers were inoculated with infectious bovine rhinotracheitis virus (IBRV) intranasally. Average daily gain from d 0 to 80 was increased (P < .10) by supplemental Cr. There was a shipping x time interaction for serum cortisol concentrations. Shipping increased (P < .02) serum cortisol on d 58, but 7 d after transport there were no effects of shipping on serum cortisol. Transportation increased (P < .05) the ratio of neutrophils to lymphocytes. Supplemental Cr did not affect rectal temperature after the IBRV challenge or the antibody response to IBRV or porcine red blood cells. Immunoglobulin G antibody response to porcine red blood cells was decreased (P < .09) by shipping. Supplemental Cr as Cr-nicotinic acid improved ADG of growing steers, regardless of whether they had been stressed by shipping. Supplemental Cr did not affect any of the immune responses that were measured. PMID- 9222856 TI - Duodenal flow of microbial nitrogen estimated from urinary excretion of purine derivatives in calves after early weaning. AB - Duodenal flow of microbial N (MN) was estimated from urinary purine derivatives to examine age-related changes in MN in male Holstein calves. In Exp. 1, endogenous purine derivatives were determined by measurement of purine derivatives in five calves fed nucleic acid-free milk replacer alone. In Exp. 2, the ratio of urinary excretion as purine derivatives to purines administered via the reticular groove was determined in three calves weaned at 5 wk of age. As a result, endogenous purine derivatives were constant at 705 mumol/(kg BW.75.d), irrespective of the amount of milk replacer, and the ratios of purine derivatives to duodenal purines were estimated to be .549, .276, .363, and .466 at wk 1, 6, 11, and 20 after weaning, respectively. Using these variables and urinary purine derivatives, the duodenal flow of MN was estimated and its relation with N balance was examined in 15 calves weaned at 5 wk of age in Exp. 3. Digestible OM was lower at wk 1 after weaning and transiently higher at wk 6. The percentage of N absorbed to N intake, N absorbed, N retained, and estimated duodenal MN were also lower at wk 1, and rapidly increased for the first 6 wk. These findings suggest that the increases in N absorbed and N retained for the first 6 wk after weaning were due to augmentation of duodenal flow of MN and dietary N that escaped ruminal degradation. PMID- 9222858 TI - Rapid communication: the first microsatellite DNA marker for marble trout, BFRO 001. PMID- 9222857 TI - Lysine deficiency in postweaned calves fed corn and corn gluten meal diets. AB - Holstein bull calves (n = 36) weaned at 6 wk of age were used in six trials to examine the response of N balance to postruminal administration of lysine with or without methionine in postweaned calves receiving diets based on corn and corn gluten meal. Calves were younger than 3 mo of age in Trials 1 and 2 but older than 3 mo in Trials 4 to 6. L-Lysine monohydrochloride was supplemented with or without DL-methionine twice daily through the reticular groove, except in Trial 4, in which N supplements were infused through duodenal cannulas. L-Glutamine was used as a nonspecific N source in every trial, and casein was a positive control in Trials 4 and 5. When daily CP intake from the diet was 3.9 g/kg BW, lysine was limiting for calves less than 11 wk of age (Trials 1 and 2) but not limiting for calves greater than 12 wk of age (Trial 3). No amino acid seemed to be limiting for calves greater than 20 wk of age (Trial 4) when daily CP intake was 4.1 g/kg BW, but lysine was limiting when CP intake was restricted to 3.0 g/kg BW when calves were more than 17 wk of age (Trial 5). However, lysine was not limiting above 18 wk of age (Trial 6) when CP intake was increased to 3.8 g/kg BW by adding urea to the diet. Results suggest that lysine may be limiting for corn and corn gluten meal diets only when ruminal microbial protein synthesis is restricted. PMID- 9222859 TI - Rapid communication: a novel DNA polymorphism of the porcine myogenin (MYOG) gene. PMID- 9222860 TI - Comment on the paper "Effects of space allocation and temperature on periparturient maternal behaviors, steroid concentrations, and piglet growth rates". PMID- 9222861 TI - Comment on the paper "Improving tenderness of normal and callipyge lambs with calcium chloride". PMID- 9222862 TI - Pathophysiology of ischemic heart disease with special reference to the degree of organic stenosis of the coronary artery. PMID- 9222863 TI - Decreased susceptibility of glycated very low density lipoproteins to lipoprotein lipase in vitro and prevention by glutathione. AB - In vitro glycation of very low density lipoprotein (VLDL) reduced the susceptibility to lipoprotein lipase (LPL) as the level of glycation increased. Addition of reduced glutathione to an incubation medium of serum and glucose interfered with glycation of serum proteins when the concentration of reduced glutathione was higher than 10 mM. At concentrations higher than 25 mM, it also significantly prevented the glycation induced reduction of fatty acid releases from VLDL by LPL. There were no such effects on glycation of serum protein and the fatty acid release from the addition of aminoguanidine. By contrast, addition of D-lysine enhanced glycation of serum proteins by glucose and further decreased fatty acid release from VLDL by LPL. From these results, it is suggested that glycation of VLDL decreases the susceptibility of VLDL to LPL. Delayed catabolism of VLDL in diabetic patients is considered partly caused by glycation of apoproteins, which renders VLDL less sensitive to LPL, in addition to the decreased LPL activity in diabetes mellitus. PMID- 9222864 TI - Relationships between subcutaneous fat and muscle distributions and serum HDL cholesterol. AB - To evaluate the relationship between human body composition and serum lipids levels, the distributions of subcutaneous adipose tissue (AT) and muscle thickness were evaluated in Japanese 449 males and 542 females, aged from 35 to 77 years. Among males, a significant positive correlation was observed between AT thicknesses and total cholesterol, and negative relationships between the AT and HDL-C as well as HDL-C/TC ratio. Among females, similar but weaker relationships were found for AT at the upper arm and trunk sites. However, the thigh AT thickness was positively correlated with HDL-C and HDL-C/TC ratio only among women. The muscle thickness in the abdomen and the thigh correlated significantly with HDL-C/TC for both sexes. Furthermore, the regional trend observed in both sexes remained significant after correction for concomitant variables such as age, tobacco and alcohol intake. We conclude that it is necessary to evaluate not only total body fat but muscle and AT thickness distributions when evaluating the relationship between body composition and serum lipids and lipoproteins. PMID- 9222865 TI - Determination of apolipoprotein E in high density lipoprotein fraction by immunofixation method and turbidimetric immunoassay after precipitation. AB - Apolipoprotein E (apoE) in high density lipoprotein (HDL) fraction (HDL-fr) was determined by the immunofixation method and turbidimetric immunoassay (TIA) after precipitation with phosphotungstic acid/MgCl2 in normolipidemic control subjects and patients with type IV hyperlipemia and hyper HDL-cholesterolemia. Immunofixation assay revealed two major bands of apoE in whole serum: one in the alpha-area and the other in the pre beta-area. ApoE in alpha-area (alpha-apoE) was identical to alpha-apoE in HDL-fr separated by ultracentrifugation but not to alpha-apoE in HDL-fr separated by precipitation (pHDL-fr). alpha-ApoE in pHDL-fr lacked the slower area of the band. Agarose column chromatography and gradient gel electrophoresis indicated that alpha-apoE belongs to early fractionated HDL, and that the precipitatable alpha-ApoE belongs to the higher molecular size HDL alpha-ApoE (%) estimated by immunofixation showed a strong positive correlation with apoE (%) in pHDL-fr. ApoE (%) in pHDL-fr was higher in case of hyper HDL cholesterolemia and lower in type IV hyperlipemia than in controls, and was inversely correlated with serum triglycerides (TG) and positively with HDL cholesterol (especially HDL2-cholesterol) in these subjects. It is suggested that the variation of apoE in pHDL-fr depends on the level of HDL2. Also, it may be suggested that apoE in pHDL-fr and precipitatable alpha-apoE belong to low and high molecular HDL2, respectively, and that apoE content in these particles is correlated with HDL2. PMID- 9222866 TI - A new approach for the detection of type III hyperlipoproteinemia by RLP cholesterol assay. AB - Type III is a remnant hyperlipoproteinemia identified by the presence of beta VLDL (remnant lipoprotein) as well as a genetic variant of apo E (apo E2/2). The RLP isolated from the serum of Type III patients by a new method we have developed, the RLPcholesterol assay, was identified as chylomicron and VLDL remnant. In addition, the RLP-C levels of the Type III patients were significantly higher than other hyperlipidemic patients with similar serum TG levels, while the ratio of TC/TG in RLP-C of both groups was not significantly different. The RLP-cholesterol assay appears to be useful for the screening and monitoring of Type III hyperlipoproteinemia when used in conjunction with the assays of serum TG level and genetic apo E isoform analysis. PMID- 9222867 TI - Change of lipoprotein (a) and coagulative or fibrinolytic parameters in diabetic patients with nephropathy. AB - Lipoprotein (a) (Lp(a)) is a plasma lipoprotein of high atherogenicity and competes with plasminogen at the site of plasminogen receptors. It is known that diabetic patients show a hypercoagulable state which might contribute to diabetic vascular complications. In the present study, mean levels of plasma Lp(a) and parameters of coagulation and fibrinolysis such as thrombin-antithrombin III complex (TAT) and alpha 2 plasmin inhibitor-plasmin complex (alpha 2PIC) were elevated in diabetic patients with nephropathy compared to healthy controls. A significant positive correlation was observed between the plasma levels of Lp(a) and alpha 2PIC (p < 0.05). Plasma levels of alpha 2PIC showed a significant positive correlation with those of TAT in the diabetic group, while there was no significant correlation observed in the non-diabetic group. The present results suggest that factors of Lp(a) and coagulation-fibrinolytic systems interacted, contributing to vascular complications in diabetic patients with nephropathy. PMID- 9222868 TI - Effects of lysosomal protease inhibitors on the degradation of acetylated low density lipoprotein in cultured rat peritoneal macrophages. AB - The effect of protease inhibitors, leupeptin and pepstatin A, on the metabolism of acetylated low density lipoprotein (acetyl-LDL) was investigated in cultured rat peritoneal macrophages and compared with that of chloroquine. While both leupeptin and pepstatin inhibited the proteolytic degradation of 125I-acetyl-LDL, a combination of both showed an additive effect. Similar to chloroquine, both protease inhibitors diminished [3H] oleate incorporation into cellular cholesteryl[3H] oleate and increased cholesterol content of macrophages. These results suggest that both thiol protease and cathepsin D participate in the physiological degradation of apolipoprotein in macrophages. The inhibition of apolipoprotein degradation seemed to have an effect on cholesterol metabolism in macrophages cultured with acetyl-LDL. PMID- 9222869 TI - Immunohistochemical and quantitative analysis of cellular and extracellular components of aortic atherosclerosis in WHHL rabbits. AB - To investigate changes in the major components of atherosclerotic lesions during the progression of this disease, we measured the lesional areas of macrophages, smooth muscle cells, collagen fibers, and extracellular lipid deposits in the aortas of WHHL rabbits. Aortic segments with lesions of various stages were stained for histological and immunohistochemical examination, and the area of each lesional component was measured by a color image analyzer. In the early fatty streaks observed in 3-month-old rabbits, macrophages were predominant in the intima and were also observed in the inner layer of the media. In the transitional lesions (fibro-fatty streaks) found in rabbits at 11 to 15 months of age, an increase in the lesional area of macrophages was prominent compared to other lesional components. Thus, macrophages may play an important role in the progression of aortic atherosclerosis at this stage. In advanced complicated lesions observed in rabbits at 20 to 24 months of age, the area of macrophages and smooth muscle cells did not increase, whereas the area of collagen fibers and extracellular lipid deposits increased. Therefore, both the disruption of foam cells and fibrosis may play an important role in the progression of atherosclerosis at this stage. PMID- 9222870 TI - Oxidized LDL induces an increase in the relative collagen synthesis of rabbit aortic smooth muscle cells. AB - Cell degeneration and collagenosis are the main features in atherosclerotic plaque. We examined the effects of human low density lipoprotein (LDL) on cultured rabbit aortic smooth muscle cells (SMCs). Copper oxidized LDL (LDL+Cu) injured the SMCs more than did native LDL. Cytotoxicity of oxidized LDL was prevented by the simultaneous addition of butylated hydroxytoluene (BHT) and ethylenediamine tetraacetic acid. Collagen synthesis increased up to 6 fold after incubation with 200 micrograms protein/ml of both native and oxidized LDL compared with that incubated in bovine serum albumin. Noncollagen protein synthesis was significantly reduced by oxidized LDL when compared to that by native LDL. Therefore, oxidized LDL increased the relative collagen synthesis (3.33%) to a greater extent than did native LDL (0.72%). By adding BHT to LDL+Cu, the elevated relative collagen synthesis was reversed due to the restoration of noncollagen protein synthesis while it also inhibited LDL peroxidation as evaluated by the formation of malondialdehyde (MDA). However, sodium MDA (up to 200 microM) did not induce either cytotoxicity or an increase of relative collagen synthesis. We therefore conclude that oxidized human LDL enhanced the relative collagen synthesis coinciding with the induction of injury in cultured aortic SMCs, however free MDA may not be the component responsible for these effects. PMID- 9222871 TI - Process of basement membrane re-formation after intimal denudation. AB - During re-endothelialization after intimal denudation induced by balloon catheter in the rat aorta, re-formation of the basement membrane (BM) was examined in samples taken 15 min, 3, 7, 14, 21 and 28 days after the balloon-induced injury. Although the luminal surface of the aortic wall was covered by round-shaped regenerating endothelial cells (ECs) by 7 days after intimal denudation, no continuous BM structure was detectable until 21 days. Periodic acid thiosemicarbazide gelatin-methenamine silver (PATSC-GMS) staining for electron microscopy and immunostaining of type IV collagen and laminin demonstrated the accumulation of BM components underneath the regenerating ECs after 14 days. The expression of type IV collagen mRNA was revealed in regenerating ECs by in situ hybridization. A continuous BM structure first appeared 21 days after intimal denudation and was almost complete by 28 days. Simultaneously, the regenerating ECs flattened and attached more closely to the BM than in earlier phases. In conclusion, we consider that the regenerating ECs produce the BM components and suggest that reorganization of the newly formed BM is important in the process of differentiation of regenerating ECs. PMID- 9222872 TI - The role of oxidized LDL in the atherogenic process. PMID- 9222873 TI - Inhibitory effect of atrial natriuretic peptide on accelerated endothelin secretion from cultured human endothelial cells. AB - This study investigated the effect of atrial natriuretic peptide (ANP) on endothelin (ET) secretion from cultured human endothelial cells. Confluent umbilical venous endothelial cells were incubated with experimental agents in multi-well plates, and the level of immunoreactive ET in the medium was measured by radioimmunoassay. There was no significant effect of ANP (10(-8), 10(-7) and 10(-6) M) on ET secretion after a 3- or 6-hour incubation. However, with 24-hour incubation, ANP significantly inhibited ET secretion from cultured human endothelial cells (control, 139.0 +/- 7.2 fmol/well; 10(-7) M, 89.4 +/- 4.7 fmol/well; 10(-7) M, 79.4 +/- 8.2 fmol/well; 10(-6) M, 71.0 +/- 10.1 fmol/well, P < 0.01). Furthermore, the addition of 8-bromo-cyclic GMP to the medium inhibited ET secretion (control, 147.2 +/- 2.9 fmol/well; 10(-5) M, 140.9 +/- 2.3 fmol/well; 10(-4) M, 143.0 +/- 1.0 fmol/well; 10(-3) M, 96.6 +/- 6.3 fmol/well, P < 0.01). These findings demonstrate that ANP inhibits accelerated ET secretion from cultured human endothelial cells, probably due to augmentation of intracellular cyclic GMP levels by ANP-activated guanylate cyclase. PMID- 9222874 TI - Existence of 7 alpha- and 7 beta-hydroperoxycholest-5-en-3 beta-ols in lipoproteins from diabetic patients and normal subjects. AB - We report evidence for the presence of 7 alpha- and 7 beta-hydroperoxycholest-5 en-3 beta-ols (cholesterol 7-hydroperoxides, Ch 7 alpha-OOH and Ch 7 beta-OOH, respectively) in human plasma lipoproteins in vivo, which had been reported to be markers of aging in rat skin. A comparative study was carried out focusing on the detection of Ch 7-OOHs in plasma lipoproteins from diabetic patients whose plasma has been suggested to be under high oxidative stress. Blood samples were collected from healthy volunteers (control) and diabetics with and without hypercholesterolemia. Ch 7-OOHs were isolated from low and high density lipoproteins (LDL and HDL, respectively) in the plasma of these subjects, identified, and determined by high-performance liquid chromatography with a chemiluminescence detector. The percent detection of Ch 7-OOHs in LDL from diabetics without hypercholesterolemia was similar to that in the control group. However, it was significantly higher in diabetics with hypercholesterolemia than in those without hypercholesterolemia. The percent detection of Ch 7-OOHs in HDL from diabetics without hypercholesterolemia was higher than both that in LDL from the same group and that in HDL from the control group. PMID- 9222875 TI - Characterization of aggregated low density lipoproteins induced by copper catalyzed oxidation. AB - Oxidation of low density lipoproteins (LDL) has been shown to lead to enhanced uptake by macrophages mediated by the scavenger receptor. In the present study, changes in LDL induced by copper-catalyzed oxidation were investigated using gel permeation chromatography (GPC), and the results were compared with several parameters of oxidized LDL (ox-LDL). When LDL at 200 micrograms/ml was oxidized with 10 microM Cu2+ at 37 degrees C for up to 24 hours, increases in thiobarbituric acid-reactive substances and electrophoretic mobility were first observed within 3 hours. An increase in fluorescence and a decrease in intact apolipoprotein B (apoB) were than observed in parallel with an increase in 125I LDL degradation by macrophages after 6 hours. Finally, LDL aggregation separated by liquid chromatography was observed after 24 hours. The aggregated and monomeric fractions of ox-LDL were analyzed and the results compared with the monomeric fraction of native LDL. Both fractions of ox-LDL contained hardly any intact apoB and showed an intense fluorescence. The electrophoretic mobility increment of aggregated ox-LDL was almost half that of monomeric ox-LDL, yet the lysine residues of aggregated ox-LDL were more extensively decreased than those of monomeric ox-LDL. Degradation of aggregated ox-LDL by macrophages showed a slightly greater increase than that of monomeric ox-LDL. GPC analysis is a useful method to estimate the LDL aggregation, and these results provide a basis to investigate the formation of aggregated LDL. PMID- 9222876 TI - Uptake of remnant like particles (RLP) in diabetic patients from mouse peritoneal macrophages. AB - To investigate whether the remnant like particles (RLP), separated from serum by an immunoaffinity gel mixture of anti-apo B-100 and apo A-I monoclonal antibodies, are relevant to the initiation or progression of atherosclerosis, the incorporation of RLP into mouse macrophages was studied using histochemical and biochemical techniques. Remnant lipoproteins such as RLP are reported to contain a large quantity of chyloniron and very low density lipoprotein (VLDL) remnants, especially in diabetic patients. The RLP separated from the sera of 32 diabetic patients were found to be predominantly taken up into macrophages harvested from mouse abdominal cavities by the staining method applying oil red O. Furthermore, using 14C-oleate to prove the uptake of lipoproteins by macrophages, the uptake of RLP-VLDL, a VLDL fraction of RLP by ultracentrifugation, was the next highest to that of the oxidized LDL, which suggests that RLP-VLDL is also aggressively taken up by macrophages. The degree of uptake of RLP-VLDL by macrophages was positively correlated with HbA1c of these diabetic patients (r = 0.556, p < 0.01), irrespective of the ways of the treatment of diabetes. In conclusion, RLP can contribute to the foaming of macrophages, which in turn may explain the acceleration of atherosclerosis in diabetic patients. PMID- 9222877 TI - Effects of LDL-apheresis on serum lipoprotein (a), C4b binding protein, protein C, protein S, and complement components. AB - The short-term effects of low-density lipoprotein (LDL) apheresis using a dextran sulfate-cellulose (DSC) column equipped with a plasma separator using a polysulfone (PS) membrane filter on the serum total cholesterol, lipoprotein(a) (Lp(a)), C4b binding protein (C4bp), protein C and protein S and complement components levels were examined in a patient with familial hypercholesterolemia (heterozygote, type IIa). PS/DSC-LDL apheresis markedly lowered the total cholesterol by 69.9 +/- 2.9%, serum Lp(a) level by 80.2 +/- 3.4%, and C4bp by 81.1 +/- 2.5% after 8 apheresis sessions. All the above parameters gradually returned to the baseline levels within 10 days. The free protein S level was not significantly changed, while the C4bp/protein S complex level was markedly decreased relative to the decrease in C4bp. The protein C level was moderately reduced by 29%. Almost all serum complements and complement co-factors as well as C4bp were moderately to markedly decreased after LDL apheresis, but returned to the initial levels within a few days. PS/DSC-LDL apheresis effectively eliminated both serum LDL and Lp(a), and did not have an adverse effect on fibrinolysis or complement cascade in the blood. PMID- 9222878 TI - Platelet volume and urinary prostanoid metabolites in non-insulin-dependent diabetes mellitus. AB - To evaluate platelet activity in patients with non-insulin-dependent diabetes mellitus (NIDDM), we measured the mean platelet volume (MPV) and 24-hour urinary excretion of 11-dehydro-thromboxane B2 (11-dTXB2) and 6-keto-prostaglandin F1 alpha (6-kPGF1 alpha), stable metabolites of thromboxane A2 and prostacyclin, respectively. The MPV of the 103 subjects in the NIDDM group were 10.72 +/- 0.82 fl for males and 10.52 +/- 1.01 fl for females (mean +/- SD), significantly higher than those of normal controls (9.95 +/- 0.75 fl for males and 9.84 +/- 0.72 fl for females). The MPV of patients with NIDDM showed positive correlations with fasting plasma glucose level and HbA1c (r = 0.234, P < 0.05; r = 0.267, P < 0.01, respectively). The urinary excretion of 11-dTXB2 was greater in the NIDDM group (7.58 +/- 4.42 micrograms/day for males and 5.65 +/- 2.38 micrograms/day for females) than in the normal controls (4.61 +/- 2.31 and 3.83 +/- 1.60, respectively), suggesting that the synthesis of thromboxane A2 by platelets may be accelerated in vivo in patients with NIDDM. The urinary 6-kPGF1 alpha was not different between the NIDDM group and normal controls among the males, but was greater in the NIDDM group among the females. As MPV showed a positive correlation (r = 0.364, P < 0.05) with urinary excretion of 11-dTXB2, MPV may be related to platelet activity. These findings suggest that the platelets of patients with NIDDM may be in a hyperactive state. PMID- 9222879 TI - Serum sialic acid concentration and atherosclerotic risk factors. AB - To clarify the significance of increased blood sialic acid in atherosclerotic cardiovascular disease, we investigated the relationship between serum sialic acid level and atherosclerotic risk factors such as serum uric acid, fasting blood glucose, systolic and diastolic blood pressure, atherogenic index, and white blood cell count. By simple regression analysis, the serum sialic acid level was found to correlate significantly with these parameters. The mean sialic acid level was significantly higher in the highest quartile for serum uric acid than in its lowest quartile and also for fasting blood glucose concentration in comparison with its other three quartiles. A tendency was noted for the mean serum sialic acid level of each quartile to become higher with an increase in the quartiles of systolic or diastolic blood pressure, atherogenic index, and white blood cell count. Multiple regression analysis showed the correlations to be significant for the relationship of serum sialic acid to atherogenic index, mean arterial pressure and white blood cell count. From these results and previous findings of higher serum sialic acid levels in smokers, patients with diabetic angiopathies, and patients with hyperlipidemia, it is suggested that serum sialic acid reflects the degree of atherosclerotic progress involving inflammation processes. PMID- 9222880 TI - Development of atherosclerosis in alloxan diabetic rats. AB - Rats with alloxan-induced diabetes developed severe atherosclerotic lesions when they were maintained on a 0.25% cholesterol diet for one year. The atheromatous changes developed at the aortic arch, appeared as early as 3 months after the start of the experiment, and increased thereafter. The diabetic rats also developed atherosclerosis when they were fed standard rat chow, but the area of the atheromatous lesion was about one tenth of that in rats fed the high cholesterol diet. Normal rats did not develop atherosclerosis even when fed the high-cholesterol diet for one year. The alloxan diabetic rats showed no increase in body weight, but developed serum glucose levels as high as 600-800 mg/dl as well as high serum cholesterol levels and lower serum HDL-cholesterol levels. The development of atherosclerosis in these rats was significantly related to an increase in the serum cholesterol/phospholipid ratio, the atherogenic index (TC HDLC/HDLC), and the serum total cholesterol level, but was not related to the serum glucose, HDL-cholesterol, triglyceride, or lipid peroxide levels. These relationships were found as early as B-16 weeks after the start of the experiment. These data suggest that the serum cholesterol/phospholipid ratio, the atherogenic index, and the total cholesterol level are important risk factors for the development of atherosclerosis in rats with alloxan diabetes. PMID- 9222881 TI - Concurrent development of hemiplegia and angina pectoris in a 46-year-old man with familial hypercholesterolemia and elevated serum Lp(a) concentrations. AB - We report a 46-year-old man with familial hypercholesterolemia who simultaneously developed angina pectoris and left hemiplegia. Angiography revealed complete tapering occlusion of the right internal carotid artery and a 75% stenosis of the right coronary artery. In addition to hypercholesterolemia, his serum Lp(a) levels were very high, with a mean (+/- SE) of 62 +/- 2 mg/dl. PMID- 9222882 TI - Cholesteryl ester accumulation in foam cells and extracellular space of atherosclerotic lesions. PMID- 9222883 TI - Immune and inflammatory mechanisms in the pathogenesis of atherosclerosis. PMID- 9222884 TI - Local expression of inflammatory cytokines in human atherosclerotic plaques. AB - Human atherosclerotic plaques are heterogeneous tissues containing a number of different cell types, including macrophages, smooth muscle, endothelial and other undefined mesenchymal-appearing cells. Significant numbers of macrophages are found in human atherosclerotic plaques and have been postulated to be a major source of growth factor production during atherogenesis. In vitro evidence suggested that macrophages synthesize PDGF and might therefore contribute to the growth of the vessel wall in atherosclerosis. However, examination of PDGF synthesis in human atheroma by in situ hybridization revealed that while smooth muscle, mesenchymal, and endothelial cells synthesize this growth factor macrophages did not. Our inability to detect PDGF mRNA in macrophages was not due to any problems with hybridization to this cell type. In situ hybridization studies on human atherosclerotic plaques have demonstrated that plaque macrophages contain many different mRNAs other than PDGF including tissue factor, factor XIII, apoprotein E, transforming growth factor beta, and tumor necrosis factor. Recent studies have indicated that macrophages may be a major source as well of another group of inflammatory cytokines which are members of the RANTES/SIS cytokine family. In situ hybridization studies on human carotid endarterectomy specimens using probes specific for the inflammatory cytokines RANTES, LD78, HIMAP, and MCP-1 revealed numerous cells containing the mRNAs encoding for these proteins (5%, 13%, 8%, and 16% of plaque cells respectively). This is in contrast to generally low level expression found in normal human arteries (< 1% of normal medial cells contain these mRNAs). Cells expressing these cytokines were often found associated with inflammatory zones in human atherosclerotic plaques. Serial section immunohistochemistry suggests that macrophages and/or T cells may synthesize these proteins. In addition to localization to macrophages MCP-1 expression was also detected in smooth muscle cells and mesenchymal-appearing cells with many of the morphological characteristics of cells previously seen to express PDGF. In vitro evidence suggests that these proteins may be chemotactic to monocytes and lymphocytes. The finding of increased expression of these mRNAs in human atheroma suggests they may play a role in monocyte trafficking into the atherosclerotic plaque. PMID- 9222885 TI - Participation of T lymphocytes and macrophages in atherogenesis. PMID- 9222886 TI - Genes expressed in monocyte/macrophage cell differentiation and atherosclerotic plaques in WHHL rabbits. PMID- 9222887 TI - Expression of c-fms on smooth muscle cells isolated from experimental arteriosclerosis. AB - In the present study, we demonstrated gene transcription of c-fms in smooth muscle cells isolated from an experimental rabbit model of arteriosclerosis (intimal smooth muscle cells), although there was no gene transcription of c-fms detected in medial smooth muscle cells. On immunocytochemical analysis, both types of smooth muscle cells similarly reacted with an antibody specific to muscle cells (HHF 35), but did not react with an antibody specific to rabbit macrophages (RAM 11). Intimal smooth muscle cells bound to acetylated LDL and resulting foam cell formation was observed. In response to M-CSF, an increased rate of cell proliferation was observed in intimal smooth muscle cells, but not in medial smooth muscle cells. These results indicated that intimal smooth muscle cells have monocyte-macrophages characteristics such as the expression of c-fms and scavenger receptor gene. PMID- 9222888 TI - Bidirectional regulation of smooth muscle cell proliferation by IFN-gamma. AB - We studied the effects of interferon gamma (IFN-gamma), a T-cell lymphokine, on the proliferation and chemotaxis of vascular smooth muscle cells (SMC). Recombinant human IFN-gamma dose-dependently inhibited the proliferation of SMC cultured in the presence of 20% fetal calf-serum. It also inhibited PDGF-induced chemotaxis of SMC. Similar concentrations of IFN-gamma induced DNA-synthesis of SMC cultured in mitogen-depleted medium for 5 days. The inhibition and the stimulation of SMC proliferation were accompanied by concomitant decrease and increase in the number of PDGF receptors. Our study indicated that IFN-gamma is a bidirectional regulator of SMC proliferation. PMID- 9222889 TI - Vascular smooth muscle cell migration and extracellular matrix. PMID- 9222891 TI - Phenotypic changes of human aortic smooth muscle cells during development and in adult. PMID- 9222890 TI - Extracellular matrix components and integrins in the control of arterial smooth muscle cell structure and function. PMID- 9222892 TI - Accumulation of vitronectin in atherosclerotic lesions where lipids deposited. PMID- 9222893 TI - Epithelial Na channels. Why all the subunits? PMID- 9222894 TI - Biophysics of a trespasser, Na+ block of Ca2+ channels. PMID- 9222895 TI - Diversity of channels generated by different combinations of epithelial sodium channel subunits. AB - The epithelial sodium channel is a multimeric protein formed by three homologous subunits: alpha, beta, and gamma; each subunit contains only two transmembrane domains. The level of expression of each of the subunits is markedly different in various Na+ absorbing epithelia raising the possibility that channels with different subunit composition can function in vivo. We have examined the functional properties of channels formed by the association of alpha with beta and of alpha with gamma in the Xenopus oocyte expression system using two microelectrode voltage clamp and patch-clamp techniques. We found that alpha beta channels differ from alpha gamma channels in the following functional properties: (a) alpha beta channels expressed larger Na+ than Li+ currents (INa+/ILi+ 1.2) whereas alpha gamma channels expressed smaller Na+ than Li+ currents (INa+/ILi+ 0.55); (b) the Michaelis Menten constants (Km of activation of current by increasing concentrations of external Na+ and Li+ of alpha beta channels were larger (Km > 180 mM) than those of alpha gamma channels (Km of 35 and 50 mM, respectively); (c) single channel conductances of alpha beta channels (5.1 pS for Na+ and 4.2 pS for Li+) were smaller than those of alpha gamma channels (6.5 pS for Na+ and 10.8 pS for Li+); (d) the half-inhibition constant (Ki) of amiloride was 20-fold larger for alpha beta channels than for alpha gamma channels whereas the Ki of guanidinium was equal for both alpha beta and alpha gamma. To identify the domains in the channel subunits involved in amiloride binding, we constructed several chimeras that contained the amino terminus of the gamma subunit and the carboxy terminus of the beta subunit. A stretch of 15 amino acids, immediately before the second transmembrane domain of the beta subunit, was identified as the domain conferring lower amiloride affinity to the alpha beta channels. We provide evidence for the existence of two distinct binding sites for the amiloride molecule: one for the guanidium moiety and another for the pyrazine ring. At least two subunits alpha with beta or gamma contribute to these binding sites. Finally, we show that the most likely stoichiometry of alpha beta and alpha gamma channels is 1 alpha: 1 beta and 1 alpha: 1 gamma, respectively. PMID- 9222896 TI - Block of N-type calcium channels in chick sensory neurons by external sodium. AB - L-type Ca2+ channels select for Ca2+ over sodium Na+ by an affinity-based mechanism. The prevailing model of Ca2+ channel permeation describes a multi-ion pore that requires pore occupancy by at least two Ca2+ ions to generate a Ca2+ current. At [Ca2+] < 1 microM, Ca2+ channels conduct Na+. Due to the high affinity of the intrapore binding sites for Ca2+ relative to Na+, addition of microM concentrations of Ca2+ block Na+ conductance through the channel. There is little information, however, about the potential for interaction between Na+ and Ca2+ for the second binding site in a Ca2+ channel already occupied by one Ca2+. The two simplest possibilities, (a) that Na+ and Ca2+ compete for the second binding site or (b) that full time occupancy by one Ca2+ excludes Na+ from the pore altogether, would imply considerably different mechanisms of channel permeation. We are studying permeation mechanisms in N-type Ca2+ channels. Similar to L-type Ca2+ channels, N-type channels conduct Na+ well in the absence of external Ca2+. Addition of 10 microM Ca2+ inhibited Na+ conductance by 95%; and addition of 1 mM Mg2+ inhibited Na+ conductance by 80%. At divalent ion concentrations of 2 mM, 120 mM Na+ blocked both Ca2+ and Ba2+ currents. With 2 mM Ba2+, the IC50 for block of Ba2+ currents by Na+ was 119 mM. External Li+ also blocked Ba2+ currents in a concentration-dependent manner, with an IC50 of 97 mM. Na+ block of Ba2+ currents was dependent on [Ba2+]; increasing [Ba2+] progressively reduced block with an IC50 of 2 mM. External Na+ had no effect on voltage-dependent activation or inactivation of the channel. These data suggest that at physiological concentrations, Na+ and Ca2+ compete for occupancy in a pore already occupied by a single Ca2+. Occupancy of the pore by Na+ reduced Ca2+ channel conductance, such that in physiological solutions, Ca2+ channel currents are between 50 and 70% of maximal. PMID- 9222897 TI - Calcium current activated by depletion of calcium stores in Xenopus oocytes. AB - Ca(2+) currents activated by depletion of Ca(2+) stores in Xenopus oocytes were studied with a two-electrode voltage clamp. Buffering of cytosolic Ca(2+) with EGTA and MeBAPTA abolished I(Cl(Ca)) and unmasked a current in oocytes that was activated by InsP(3) or ionomycin in minutes and by thapsigargin or the chelators themselves over hours. At -60 mV in 10 mM extracellular CaCl(2), the current was typically around -90 or -160 nA in oocytes loaded with EGTA or MeBAPTA, respectively. This current was judged to be a Ca(2+)-selective current for the following reasons: (a) it was inwardly rectifying and reversed at membrane potentials usually more positive than +40 mV; (b) it was dependent on extracellular [CaCl(2)] with K(m) = 11.5 mM; (c) it was highly selective for Ca(2+) against monovalent cations Na(+) and K(+), because replacing Na(+) and K(+) by N-methyl-d-glucammonium did not reduce the amplitude or voltage dependence of the current significantly; and (d) Ca(2+), Sr(2+), and Ba(2+) currents had similar instantaneous conductances, but Sr(2+) and Ba(2+) currents appeared to inactivate more strongly than Ca(2+). This Ca(2+) current was blocked by metal ions with the following potency sequence: Mg(2+) << Ni(2+) approximately Co(2+) approximately Mn(2+) < Cd(2+) << Zn(2+) << La(3+). It was also inhibited by niflumic acid, which is commonly used to block I(Cl(Ca)). PMA partially inhibited the Ca(2+) current, and this effect was mostly abolished by calphostin C, indicating that the Ca(2+) current is sensitive to protein kinase C. These results are the first detailed electrophysiological characterization of depletion activated Ca(2+) current in nondialyzed cells. Because exogenous molecules and channels are easy to introduce into oocytes and the distortions in measuring I(Cl(Ca)) can now be bypassed, oocytes are now a superior system in which to analyze the activation mechanisms of capacitative Ca(2+) influx. PMID- 9222898 TI - Calcium signaling in transgenic mice overexpressing cardiac Na(+)-Ca2+ exchanger. AB - We have produced transgenic mice which overexpress cardiac Na(+)-Ca2+ exchange activity. Overexpression has been assessed by Western blot, Northern blot, and immunofluorescence. Functional overexpression was analyzed using membrane vesicles and isolated ventricular myocytes. In whole cell clamped myocytes dialyzed with 0.1-0.2 mM Fura-2, the magnitude of ICa and Ca2+i-transient triggered by ICa or caffeine were not significantly different in transgenic vs. control myocytes. In transgenic myocytes, activation of ICa, however, was followed by a large slowly inactivating transient inward current representing INa Ca. This current depended on Ca2+ release as it was abolished when sarcoplasmic reticulum (SR) Ca2+ was depleted using thapsigargin. Cai-transients triggered by rapid application of 5 mM caffeine, even though equivalent in control and transgenic myocytes, activated larger INa-Ca (approximately 5 pA/pF at -90 mV) in transgenic vs. control myocytes (1.5 pA/pF). The decay rate of caffeine-induced Ca2+i-transient and INa-Ca was 2.5 times faster in transgenic than in control myocytes. 5 mM Ni2+ was equally effective in blocking INa-Ca in control or transgenic myocytes. In 9 out of 26 transgenic myocytes, but none of the controls, Ca2+ influx via the exchanger measured at +80 mV caused a slow rise in [Ca2+]i triggering rapid release of Ca2+ from the SR, SR Ca2+ release triggered by the exchanger at such potentials was accompanied by activation of transient current in the inward direction. In 2 mM Fura-2-dialyzed transgenic myocytes caffeine-triggered Cai-transients failed to activate INa-Ca even though the kinetics of inactivation of ICa slowed significantly in caffeine-treated myocytes. In 0.1 mM Fura-2-dialyzed transgenic myocytes 100 microM Cd2+ effectively blocked ICa and suppressed Cai-transients at -10 or +50 mV. Our data suggests that in myocytes overexpressing the exchanger, the content of intracellular Ca2+ pools and the signaling of its release by the Ca2+ channel vis a-vis the Na(+)-Ca2+ exchanger were not significantly altered despite an up to ninefold increase in the exchanger activity. We conclude that the exchanger remains functionally excluded from the Ca2+ microdomains surrounding the DHP/ryanodine receptor complex. PMID- 9222899 TI - A highly temperature-sensitive proton current in mouse bone marrow-derived mast cells. AB - Proton (H+) conductive pathways are suggested to play roles in the regulation of intracellular pH. We characterized temperature-sensitive whole cell currents in mouse bone marrow-derived mast cells (BMMC), immature proliferating mast cells generated by in vitro culture. Heating from 24 to 36 degrees C reversibly and repeatedly activated a voltage-dependent outward conductance with Q10 of 9.9 +/- 3.1 (mean +/- SD) (n = 6). Either a decrease in intracellular pH or an increase in extracellular pH enhanced the amplitude and shifted the activation voltage to more negative potentials. With acidic intracellular solutions (pH 5.5), the outward current was detected in some cells at 24 degrees C and Q10 was 6.0 +/- 2.6 (n = 9). The reversal potential was unaffected by changes in concentrations of major ionic constituents (K+, Cl-, and Na+), but depended on the pH gradient, suggesting that H+ (equivalents) is a major ion species carrying the current. The H+ current was featured by slow activation kinetics upon membrane depolarization, and the activation time course was accelerated by increases in depolarization, elevating temperature and extracellular alkalization. The current was recorded even when ATP was removed from the intracellular solution, but the mean amplitude was smaller than that in the presence of ATP. The H+ current was reversibly inhibited by Zn2+ but not by bafilomycin A1, an inhibitor for a vacuolar type H(+)-ATPase. Macroscopic measurements of pH using a fluorescent dye (BCECF) revealed that a rapid recovery of intracellular pH from acid-load was attenuated by lowering temperature, addition of Zn2+, and depletion of extracellular K+, but not by bafilomycin A1. These results suggest that the H+ conductive pathway contributes to intracellular pH homeostasis of BMMC and that the high activation energy may be involved in enhancement of the H+ conductance. PMID- 9222900 TI - Regulation of Ca2+ release by InsP3 in single guinea pig hepatocytes and rat Purkinje neurons. AB - The repetitive spiking of free cytosolic [Ca2+] ([Ca2+]i) during hormonal activation of hepatocytes depends on the activation and subsequent inactivation of InsP3-evoked Ca2+ release. The kinetics of both processes were studied with flash photolytic release of InsP3 and time resolved measurements of [Ca2+]i in single cells. InsP3 evoked Ca2+ flux into the cytosol was measured as d[Ca2+]i/dt, and the kinetics of Ca2+ release compared between hepatocytes and cerebellar Purkinje neurons. In hepatocytes release occurs at InsP3 concentrations greater than 0.1-0.2 microM. A comparison with photolytic release of metabolically stable 5-thio-InsP3 suggests that metabolism of InsP3 is important in determining the minimal concentration needed to produce Ca2+ release. A distinct latency or delay of several hundred milliseconds after release of low InsP3 concentrations decreased to a minimum of 20-30 ms at high concentrations and is reduced to zero by prior increase of [Ca2+]i, suggesting a cooperative action of Ca2+ in InsP3 receptor activation. InsP3-evoked flux and peak [Ca2+]i increased with InsP3 concentration up to 5-10 microM, with large variation from cell to cell at each InsP3 concentration. The duration of InsP3 evoked flux, measured as 10-90% risetime, showed a good reciprocal correlation with d[Ca2+]i/dt and much less cell to cell variation than the dependence of flux on InsP3 concentration, suggesting that the rate of termination of the Ca2+ flux depends on the free Ca2+ flux itself. Comparing this data between hepatocytes and Purkinje neurons shows a similar reciprocal correlation for both, in hepatocytes in the range of low Ca2+ flux, up to 50 microM. s-1 and in Purkinje neurons at high flux up to 1,400 microM. s-1. Experiments in which [Ca2+]i was controlled at resting or elevated levels support a mechanism in which InsP3-evoked Ca2+ flux is inhibited by Ca2+ inactivation of closed receptor/channels due to Ca2+ accumulation local to the release sites. Hepatocytes have a much smaller, more prolonged InsP3-evoked Ca2+ flux than Purkinje neurons. Evidence suggests that these differences in kinetics can be explained by the much lower InsP3 receptor density in hepatocytes than Purkinje neurons, rather than differences in receptor isoform, and, more generally, that high InsP3 receptor density promotes fast rising, rapidly inactivating InsP3-evoked [Ca2+]i transients. PMID- 9222901 TI - Mutation in the M1 domain of the acetylcholine receptor alpha subunit decreases the rate of agonist dissociation. AB - We describe the kinetic consequences of the mutation N217K in the M1 domain of the acetylcholine receptor (AChR) alpha subunit that causes a slow channel congenital myasthenic syndrome (SCCMS). We previously showed that receptors containing alpha N217K expressed in 293 HEK cells open in prolonged activation episodes strikingly similar to those observed at the SCCMS end plates. Here we use single channel kinetic analysis to show that the prolonged activation episodes result primarily from slowing of the rate of acetylcholine (ACh) dissociation from the binding site. Rate constants for channel opening and closing are also slowed but to much smaller extents. The rate constants derived from kinetic analysis also describe the concentration dependence of receptor activation, revealing a 20-fold shift in the EC50 to lower agonist concentrations for alpha N217K. The apparent affinity of ACh binding, measured by competition against the rate of 125I-alpha-bungarotoxin binding, is also enhanced 20-fold by alpha N217K. Both the slowing of ACh dissociation and enhanced apparent affinity are specific to the lysine substitution, as the glutamine and glutamate substitutions have no effect. Substituting lysine for the equivalent asparagine in the beta, epsilon, or delta subunits does not affect the kinetics of receptor activation or apparent agonist affinity. The results show that a mutation in the amino-terminal portion of the M1 domain produces a localized perturbation that stabilizes agonist bound to the resting state of the AChR. PMID- 9222902 TI - Role of transmembrane segment S5 on gating of voltage-dependent K+ channels. AB - The cytoplasmic half of S5 (5'S5) has been identified as part of the inner mouth of the pore based on evidence that mutations in this region greatly alter single channel conductance, 4-aminopyridine (4-AP) block and the rate of channel closing upon repolarization (deactivation). The latter effect, suggestive of a role for 5'S5 in channel gating was investigated in the present study. The biophysical properties of chimeric channels, in which the 5'S5 regions were exchanged between two host channels (Kv2.1 and Kv3.1) that differ in 4-AP sensitivity and deactivation rate, were examined in a Xenopus oocyte expression system. Exchange of 5'S5 between Kv2.1 and Kv3.1 confers steady-state voltage dependence of activation and rates of channel deactivation similar to those of the donor channel. The involvement of voltage-dependent gating was confirmed by the observation that exchanging the 5'S5 segment of Kv2.1 with that of Kv3.1 confers a change from slow to fast deactivation kinetics by accelerating the decay of off gating charge movement. We suggest that a conformational change that extends from the voltage-sensor in S4 to the region of the pore lined by S5 regulates the stability of the open state. Therefore, the cytoplasmic end of S5, in addition to forming part of the conduction pathway near the inner mouth of the pore, also participates in the conformational rearrangements associated with late steps in channel activation and early steps in deactivation. PMID- 9222903 TI - How does the W434F mutation block current in Shaker potassium channels? AB - The mutation W434F produces an apparently complete block of potassium current in Shaker channels expressed in Xenopus oocytes. Tandem tetrameric constructs containing one or two subunits with this mutation showed rapid inactivation, although the NH2-terminal inactivation domain was absent from these constructs. The inactivation showed a selective dependence on external cations and was slowed by external TEA; these properties are characteristic of C-type inactivation. Inactivation was, however, incompletely relieved by hyperpolarization, suggesting the presence of a voltage-independent component. The hybrid channels had near normal conductance and ion selectivity. Single-channel recordings from patches containing many W434F channels showed occasional channel openings, consistent with open probabilities of 10(-5) or less. We conclude that the W434F mutation produces a channel that is predominantly found in an inactivated state. PMID- 9222904 TI - The hand and the eye. AB - A variety of systemic diseases may affect both the hand and eye, leading to profound sensory and functional deficits. The aetiology of some of the most common conditions is reviewed, with an explanation of their manifestations in the hand and eye. Recognition of the association between a hand condition and ocular pathology may aid in diagnosis of a systemic disorder or allow early detection and prevention of ocular disease and loss of vision. PMID- 9222905 TI - A new type of "bioartificial" nerve graft for bridging extended defects in nerves. AB - In the rat sciatic nerve, a gap of around 10 mm in nerve continuity seems to be the maximal distance which can be successfully repaired by silicone tubes. In this study we tested if a new artificial nerve graft, composed of eight polyamide filaments (diameter 250 microns) placed inside silicone tubes (1.8 mm inner diameter), could be used to bridge an extended gap (15 mm) in rat sciatic nerve. Silicone tubes containing eight polyamide sutures were found to support regeneration across such a gap. After 4 weeks sensory fibres had bridged the gap and grown into the distal nerve segment as revealed by a positive pinch reflex test as well as positive staining for neurofilaments in the distal nerve segment. Myelinated axons could be observed in the tissue matrix formed in between and peripheral to the synthetic filaments along the whole length of the tube. In contrast, when silicone tubes without filaments were used to bridge the 15 mm gap, the tubes contained only fluid or in two cases a thin tissue strand. No positive pinch reflex response was elicited in the nerve segment distal to such a tube. We conclude that the new artificial nerve graft can be used to support regeneration across extended gaps in nerves. PMID- 9222906 TI - Nerve xenograft transplantation. Immunosuppression with FK-506 and RS-61443. AB - An experimental model has been developed to study the potential transplantation of nerve xenografts using the newer immunosuppressive agents RS-61443 and FK-506. Sciatic nerve grafts of 2 cm were transplanted from donor Golden Syrian hamsters into a 0.5 cm gap in the sciatic nerve of recipient Lewis rats. Walking track analysis, somatosensory evoked potentials and histology demonstrated improved regeneration across the nerve xenografts that had been immunosuppressed with RS 61443 and FK-506 compared with non-immunosuppressed controls, but the function never approached that seen in control isografts. Regeneration across nerve xenografts immunosuppressed with FK-506 was better than xenografts immunosuppressed with RS-61443. PMID- 9222907 TI - Cold intolerance following peripheral nerve injury. Natural history and factors predicting severity of symptoms. AB - Cold intolerance can be severe and debilitating following injury to the hand. Little is known about its natural history and factors predicting symptom severity. We looked retrospectively at upper limb peripheral nerve injuries over a 12-year period. Information was obtained using a patient questionnaire and patient records. The incidence of cold intolerance was 83%. In 48% the onset of symptoms was within 1 month of the initial injury. At a mean follow-up of 51 months improved symptoms were reported by 21%, while 18% deteriorated. Patients were more likely to develop cold intolerance if they smoked and less likely if they suffered a sharp injury. A score defining the severity of cold induced symptoms, based on the information collected, was calculated for each patient. Significantly increased severity was associated with complete nerve division, median and ulnar nerve division and an associated vessel injury. Symptom improvement was significantly more likely in non-smokers and a deterioration most likely with a high severity score. PMID- 9222908 TI - Local symptoms after open carpal tunnel release. A randomized prospective trial of two incisions. AB - We report a randomized trial of two skin incisions for carpal tunnel decompression, namely a standard incision and an ulnar L incision. We looked particularly at the resolution of local symptoms namely pillar pain and scar sensitivity. There were 47 patients in the trial. No difference was found in pillar pain between the two incisions, but one had a lower incidence of scar sensitivity. These results give a baseline for comparison of local postoperative symptoms following open release with those following endoscopic release. PMID- 9222909 TI - Carpal tunnel decompression under local anaesthetic and tourniquet control. AB - A postal survey within the North West Region (UK) revealed that 66% of the consultant orthopaedic surgeons did not use local anaesthesia routinely for carpal tunnel decompression. This prospective study was set up to assess the effectiveness, safety and patient tolerance of performing this procedure using local anaesthesia and upper arm tourniquet control. Eight-six carpal tunnel decompressions were performed on 75 consecutive and unselected patients with confirmed carpal tunnel syndrome over a 6-month period. Completed questionnaires were obtained before discharge. None or only slight discomfort was experienced for all aspects of the operation in at least 94%. None of the patients reported severe and unbearable discomfort. At review, 3 months postoperatively, all patients with the exception of two reported complete resolution of preoperative symptoms. The use of local anaesthesia and a tourniquet is safe, effective and well-tolerated in carpal tunnel decompressions. PMID- 9222910 TI - Touch allodynia following endoscopic (single portal) or open decompression for carpal tunnel syndrome. AB - We investigated if single-portal endoscopic carpal tunnel decompression equipment (Agee, 3M, USA) would cause increased carpal tunnel pressure during the release and if endoscopic release would reduce postoperative touch allodynia. Measurements on cadavers of the pressure produced during endoscopic release showed similar pressures to those produced during maximal range of motion. One hundred patients underwent either open or endoscopic decompressions. Twenty normal individuals served as controls. At 1 month after surgery both groups had significant allodynia compared with the controls, but at 3 months the endoscopic group had returned to normal though the open group was still significantly abnormal. The reported endoscopic release may therefore be of particular advantage to patients who would seriously be disadvantaged if postoperative touch allodynia should develop. The Agee endoscope is unlikely to cause disturbance of the nerve function due to increased carpal pressure during the release. PMID- 9222911 TI - Martin-Gruber connections in South Africa. An anatomical study. AB - One hundred and twelve forearms in 56 preserved cadavers were dissected to assess the incidence of Martin-Gruber connections in a population in South Africa. The connections were found in 13 cadavers (23%) and one was bilateral. There were no significant racial or sexual differences in the incidence. The course of Martin Gruber connections and their anatomical relationship with the ulnar artery are illustrated. PMID- 9222912 TI - ADCON-T/N reduces in vivo perineural adhesions in a rat sciatic nerve reoperation model. AB - Excessive perineural scarring may affect the result of peripheral nerve surgery. The ability of a novel implant material (ADCON-T/N) to prevent this complication was tested in 38 rats. Four weeks after a bilateral sciatic nerve external neurolysis, a secondary bilateral lysis of the adhesions was performed; ADCON-T/N was locally implanted at one side, while the contralateral side was left untreated. Four or 8 weeks later, perineural adhesions were dissected in 24 animals and graded blindly. Significantly fewer perineural adhesions were found in ADCON-T/N treated nerves compared with controls at both 4 and 8 weeks. Residual implant material or adverse effects were not observed at either time. Histological examination of the neurolysis sites in another 14 animals confirmed these findings at both time intervals. This study shows that ADCON-T/N is effective in inhibiting perineural adhesions, is resorbed within 4 weeks and is well tolerated. PMID- 9222913 TI - Contralateral transplantation of a finger for restoration of thumb function. AB - Transplantation of a finger from the contralateral hand for thumb reconstruction is seldom done because of possible psychological problems for the patient. We present two cases in which a previously damaged index finger of the contralateral hand was transplanted. In both patients the metacarpophalangeal joint of the index finger replaced that of the thumb. A powerful pinch to the fingers was achieved and the appearance of both the donor and the recipient hands was considerably improved. PMID- 9222914 TI - Composite graft replacement of digital tips. 1. Before 1850 and after 1950. AB - The successes of composite grafting of fingertips in the early years of plastic surgery have been repeated in the few studies of this treatment which have been reported during the last 50 years. The early and recent history of this subject are reviewed in the light of the increasingly pessimistic view of composite graft replacement of fingertips taken by recent reviewers. PMID- 9222915 TI - Composite graft replacement of digital tips. 2. A study in children. AB - This study investigated the outcome of composite graft replacement of 50 amputated digital tips in 50 children over a period of 3 years and 6 months. Eleven of 18 tips (61%) which were replaced within 5 hours survived completely while none of 32 digital tips replaced after 5 hours survived completely. This difference was highly significant. The mean delay time between amputation and replacement in the successful group was 3.9 hours and in the others was 7.2 hours. This difference was also statistically significant. The implications of the findings of this series to the use of this treatment are discussed. PMID- 9222917 TI - Limited microsurgical arteriolysis for complications of digital vasospasm. AB - Twenty-two digits in seven patients with chronic digital vasospasm were surgically treated after failed medical management. All patients complained of severe ischaemic pain. Chronic digital tip ulceration was present in seven digits and dry gangrene in one. Following surgical microarteriolysis all digital ulcers healed completely. Severe digital ischaemic pain was significantly improved in all digits and completely resolved in 19 of 22 digits. PMID- 9222916 TI - Very distal finger amputations: replantation or "reposition-flap" repair? AB - Management of very distal finger amputations is still controversial. Successful replantation results in an almost normal finger but is not without problems, such as technical difficulty, risk of failure and cost. "Reposition-flap" repair is a simpler procedure: it consists of distal bone and nail bed "graft-reposition" and pulp reconstruction by a flap. We compare ten successful replantations and six reposition-flap reconstructions. Replantation has several advantages over reposition-flap repair in terms of less finger shortening, longitudinal nail curvature, absence of PIP flexion contracture and shorter time off work. The results of reposition-flap repair are less satisfactory, but it is nevertheless a useful alternative when replantation is impossible or has failed. PMID- 9222919 TI - Early active mobilization after second stage flexor tendon grafts. AB - We assessed the results of nine two-stage tendon reconstructions. The tendon graft was the ipsilateral palmaris longus tendon inserted into a tunnel which had been previously created by a silicone spacer. Early active mobilization was commenced 48 hours after surgery according to a previously described protocol (Small et al, 1989). Using the grading system of Kleinert and Verdan (1983) the results were: one excellent, two good, five fair and one poor. Using the Buck Gramcko et al (1976) grading system there were three excellent, two good, two satisfactory and two poor results. There were no cases of tendon graft rupture or dehiscence of the junction between tendon and graft. PMID- 9222918 TI - The dorsal middle phalangeal finger flap. Mid-term results of 43 cases. AB - The dorsal middle phalangeal finger (DMF) flap is a (neuro)vascular island flap based on one palmar proper digital artery, its venae comitantes (and/or a separate dorsal vein) and the dorsal branch(es) of the palmar digital nerve. The main nerve supply of the donor finger is left undisturbed. The flap may be raised on a short antegrade, long antegrade or a retrograde pedicle, and used as a free, arterial and/or venous flow-through or neurovascular flap. In a prospective study (mean follow-up of 50 months), the results of 43 DMF flaps were analysed. All flaps survived, retained patency of their vascular pedicles and fulfilled their goals. Neurovascular flaps provided sensate coverage at the S3+ level with static 2-point discrimination values of about 10 mm. Dissection between the proper digital nerve and the rest of the neurovascular bundle induced a 5% incidence of cold intolerance and a 12% occurrence of S3+ hypaesthesia. Advantages, drawbacks and indications of DMF flaps are outlined. PMID- 9222920 TI - Loss of flexor pollicis longus function after plating of the radius. Report of six cases. AB - Six cases of loss of flexor pollicis longus function after plating of a radius fracture are presented. The exact aetiology of the postoperative deficit is uncertain, but is probably a traction neuropraxia of the anterior interosseous nerve branches to the flexor pollicis longus. All six patients had full recovery within 5 months. An initial conservative approach is recommended if this complication is encountered. PMID- 9222921 TI - Partial lacerations of flexor tendons in children. Primary repair versus conservative treatment. AB - We compared the outcome of 17 partially lacerated (less than 75% of cross sectional area) flexor tendons in children treated by surgical repair to that of 19 tendons treated conservatively by early mobilization. The outcome of both groups was similarly favourable. No complications, such as triggering or complete tendon tear, were found in either group. We advocate early mobilization in children in whom a partial division of the flexor tendon is diagnosed clinically. Exploration should be carried out only in doubtful cases to exclude complete division of the tendon. PMID- 9222922 TI - Spontaneous rupture of the flexor pollicis longus tendon on a sesamoid bone. PMID- 9222923 TI - Flexor tendon ruptures caused by an intraosseous ganglion of the hook of the hamate. PMID- 9222924 TI - Subcutaneous extensor tendon rupture associated with calcium pyrophosphate dihydrate crystal deposition disease of the wrist. AB - We report a patient with rupture of the ring extensor tendon associated with calcium pyrophosphate dihydrate crystal deposition disease of the wrist. Crystal deposits were noted in synovium and the triangular fibrocartilage complex. Histological examination revealed chronic synovitis with foreign body giant cell reaction to crystals. The cause of the tendon rupture was synovitis due to crystal deposition. PMID- 9222925 TI - The EFFUL (Evaluation of Function in the Flail Upper Limb) system. A ranking score system to measure improvement achieved by surgical reconstruction and rehabilitation. AB - The EFFUL (Evaluation of Function of the Flail Upper Limb) system measures in numerical terms the improvement, achieved through reconstructive surgery and hand therapy, in patients with brachial plexus injuries. The EFFUL system measures practical, everyday activities performed with the shoulder, elbow, forearm, wrist and hand. The ranking system is based upon a classification of function, with a hierarchy of increasing functional demands until normal function has been achieved. These activities focus on two-handed coordination and hand dominance. The score is plotted on a star histogram known as the patient's profile, and shows the preoperative score achieved in the first evaluation, and the postoperative score achieved in the second evaluation. The shaded area between the two plotted points clearly demonstrates any improvement in function obtained by surgery and rehabilitation. It also demonstrates the interrelationship between the various regions: improvement in function in one region often results in improvement in other regions. PMID- 9222926 TI - Use of a bone block graft from the iliac crest with rigid fixation to correct diaphyseal defects of the radius and ulna. AB - We report our experience with the use of a bone block graft from the iliac crest to correct diaphyseal defects of the forearm bones. The technique was used in 12 patients (ten men and two women, average age 29 years) for defects resulting mainly from either closed or compound fractures, which later developed infection and bone tissue loss. The average dimensions of the graft required to correct the defect was 3.5 x 1.8 cm. The graft application was combined with rigid fixation with an AO 3.5 mm DCP plate, permitting early active motion. The graft incorporated without any additional grafting procedure in ten cases within 17.2 weeks on average. The most frequent complication was infection (four cases), controlled by means of debridement, cleansing and antibiotics. A comparative analysis of the immediate and final radiographs of the graft showed an average 30% loss of bone mass despite integration. We conclude that the technique of bone block grafting to correct diaphyseal defects of the radius or ulna is relatively easy to carry out and has a high success rate. PMID- 9222927 TI - Cardiac arrhythmia caused by a Kirschner wire inside the heart. An unusual complication of finger osteosynthesis. AB - A 45-year-old woman with no previous history of cardiac disease woke up one morning with an irregular heartbeat and fatigue. An electrocardiogram showed atrial fibrillation and plain chest radiographs revealed the presence of a metallic pin at the position of the heart. A 24 mm-long metallic pin was removed by open thoracic surgery from within the right ventricle of the heart. Postoperative examination of the pin showed it to be one of the 0.8 mm Kirschner wires that had been used for finger osteosynthesis in her left hand 31 months previously. PMID- 9222928 TI - Surgical management of eosinophilic fasciitis of the upper extremity. AB - Eosinophilic fasciitis is a rare inflammatory disease associated with peripheral eosinophilia, hyper-gammaglobulinaemia and contractures of any joint in the upper extremity. Although conservative treatments are generally advocated, this study reports the results of surgical intervention. Four patients aged from 20 to 48 years underwent fasciectomy followed by oral administration of prednisolone. All presented with contractures of digits, wrist, or elbow due to eosinophilic fasciitis in the upper extremities. Despite one case that required a second operation for recurrence, all patients regained the range of motion of the affected joints a few weeks after surgery. The recovery was much sooner than in previously reported cases treated conservatively, suggesting that surgical management of eosinophilic fasciitis is effective in alleviating symptoms quickly and allows patients to resume activities of daily living sooner. PMID- 9222929 TI - Peripheral gangrene associated with fulminating meningococcal septicaemia. Is early escharotomy indicated? AB - We present a case of a 3-month-old boy presenting with fulminating meningococcal septicaemia associated with extensive peripheral gangrene requiring amputation of three limbs. The surgical management options and the role of early fasciotomy are discussed. PMID- 9222930 TI - Biceps rupture in a patient on long-term anticoagulation leading to compartment syndrome and nerve palsies. PMID- 9222931 TI - False aneurysm and brachial plexus palsy complicating a proximal humeral exostosis. AB - An unusual case is described in which a false aneurysm of the brachial artery secondary to an exostosis of the proximal humerus caused a compressive lesion of the brachial plexus. Surgical treatment of the exostosis and the false aneurysm relieved the symptoms. PMID- 9222932 TI - Traumatic aneurysm of a common digital artery. AB - We describe a false aneurysm of a common digital artery. Diagnosis and choice of treatment were largely determined by ultrasonography which proved to be a reliable and easy to use diagnostic tool. PMID- 9222933 TI - Radial artery injury in association with fractures of the trapezium. AB - Three cases of trapezial fractures in which the radial artery was injured are reported. The injuries occurred during the initial trauma in one, when internal fixation was done in the second, and during hardware removal in the third. Six cadaveric wrists were dissected to study the relationship between the course of the artery and the trapezium. PMID- 9222934 TI - Candida infection of a silicone metacarpophalangeal arthroplasty. AB - Fungal infections following joint arthroplasty are extremely rare. Only 16 cases of Candida prosthetic infections have been reported, involving the hip, knee or shoulder joints. We report a case of a silicone metacarpophalangeal joint replacement complicated by a Candida albicans infection. PMID- 9222935 TI - Treatment of scaphoid nonunions with a vascularized bone graft based on the first dorsal metacarpal artery. AB - Four patients with chronic nonunion of the scaphoid were treated by a vascularized bone graft based on first dorsal metacarpal artery. The mean duration of the nonunion was 28.5 months (range 12-48 months). There was avascular necrosis in all patients confirmed by magnetic resonance imaging (MRI). None of the patients had previous attempts at surgical reconstruction. Two fractures were localized at the waist one in the distal part and one at the proximal pole. Osseous union of the scaphoid was confirmed by X-ray in all patients in an average of 2.1 months. We recommend this technique for the treatment of established scaphoid pseudoarthrosis with avascular necrosis since it is associated with minimal morbidity and predictable good results. PMID- 9222936 TI - Hand over glove. PMID- 9222937 TI - A review of mutagenesis studies of angiotensin II type 1 receptor, the three dimensional receptor model in search of the agonist and antagonist binding site and the hypothesis of a receptor activation mechanism. AB - OBJECTIVE: To seek the mechanism whereby agonists, competitive antagonists and insurmountable antagonists affect the receptor function differently, by reviewing recent mutagenesis studies of angiotensin II type 1 receptor (AT1) in which the binding of the agonist and antagonists and receptor signaling were affected. AT1 RECEPTOR STRUCTURE AND LIGAND BINDING SITES: We built a model of seven transmembrane spanning domains of the AT1 receptors using bacteriorhodopsin as a template. The carboxy terminal of angiotensin II binds to Lys199 in transmembrane domain 5, whereas the guanidinium group of Arg2 binds to Asp281 in transmembrane domain 7. Results of studies using mutagenesis supporting proposed ligand-docking models are discussed. HYPOTHESIS FOR THE LIGAND-INDUCED RECEPTOR SIGNALING MECHANISM: We submit a set of hypotheses for a mechanism whereby the ligand binding induces changes in the receptor conformation by the rotation of transmembrane helices as the initial event for the subsequent activation of a G protein. In this mechanism antagonists are not capable of rotating the helices but agonists are able to do so, which results in the formation of a hydrogen bond between Asp74 in transmembrane domain 2 and Tyr292 in transmembrane domain 7. This mechanism also provides plausible explanation for the activation of monoamine receptors. COMPETITIVE AND INSURMOUNTABLE ANTAGONISTS: Competitive antagonists share the same binding sites with agonists, but insurmountable antagonists do not, and binding of the latter does not preclude agonist binding, for example, to Asp281. CONCLUSION: This hypothesis of the intrareceptor signaling mechanism and the receptor model indicate that some amino acid residues essential for the signaling play their roles in the intrareceptor activation mechanism, whereas others participate directly in ligand binding. PMID- 9222938 TI - Hpa II polymorphism of the atrial natriuretic peptide gene and the blood pressure response to salt intake in normotensive men. AB - OBJECTIVE: To test the hypothesis that the Hpa II variant of the atrial natriuretic peptide gene (ANP), which has been reported to be more common among black hypertensives than it is among normotensive controls, is related to the response of blood pressure to salt intake in normotensive Caucasians. METHODS: One hundred and three young (aged 19-35 years) male volunteers were fed a low salt diet (20 mmol NaCl/day) for 2 weeks and a supplement of either 200 mmol NaCl/day (slow sodium) or placebo for 1 week each in a randomized double-blind cross-over order. Salt sensitivity was defined as a significant (P < 0.05) decrease in resting mean arterial blood pressure by > 3 mmHg under the low-salt diet. The genotype was determined by amplification of genomic DNA extracted from peripheral leukocytes and subsequent digestion of the amplicon with Hpa II restriction enzyme. RESULTS: According to the above definition, 27 subjects were salt sensitive. There were no significant differences in age, body-mass index and waist:hip ratio between the salt-sensitive and salt-resistant groups. Only salt sensitive subjects displayed a significantly higher blood pressure under the high salt diet (increase in mean arterial pressure 5.6 +/- 2.4 mmHg, P < 0.001). The prevalence of the ANP-Hpa II wild-type (w) allele did not differ between the salt sensitive (qw = 1.0, qm = 0) and the salt-resistant group (qw = 0.96, qm = 0.04). Furthermore, the salt-induced response of blood pressure did not differ between homozygotes (ww) and heterozygotes (wm). CONCLUSIONS: Our findings do not support the hypothesis that the ANP-Hpa II polymorphism is a marker for salt sensitivity in young Caucasian normotensives. PMID- 9222940 TI - Blunted parasympathetic modulation in salt-sensitive patients with essential hypertension: evaluation by power-spectral analysis of heart-rate variability. AB - OBJECTIVE: To evaluate autonomic nervous function by power-spectral analysis of heart-rate variability in salt-sensitive and non-salt-sensitive patients with essential hypertension under the conditions of low and high salt intakes. DESIGN AND METHODS: The blood pressures, heart rates, and electrocardiogram R-R intervals of 20 hypertensive patients were measured at intervals of 30 min during a 24 h period using a portable recorder (TM-2425) on the last day of the high- (250 mmol NaCl/day) and low-salt (25 mmol NaCl/day) diet periods. The patients whose 24 h average mean blood pressures were increased by more than 10% by the high salt intake were defined as salt-sensitive (n = 10); the other patients were considered non-salt-sensitive (n = 10). Power-spectral analysis of R-R intervals was performed to obtain the low-frequency component (0.05-0.15 Hz) and the high frequency component (0.15-0.40 Hz). RESULTS: The average 24 h blood pressure in the salt-sensitive patients was increased by the high salt intake [by 19.1 +/- 2.0/9.1 +/- 0.8 mmHg (mean +/- SEM)], whereas the heart rate did not change. In contrast, the increase in 24 h blood pressure in the non-salt-sensitive patients caused by the high salt intake was not significant and the heart rate was decreased significantly by the high salt intake (by 5.9 +/- 1.4 beats/min). The high-salt diet increased significantly the high-frequency component and decreased the low-frequency:high-frequency component ratio both during the daytime and during the night-time for the non-salt-sensitive patients. In contrast, the high frequency component and the night-time low-frequency: high-frequency component ratio of the salt-sensitive patients did not respond to dietary salt manoeuvres. CONCLUSIONS: Responses of the parasympathetic and sympathetic nervous systems to dietary salt manoeuvres were blunted in salt-sensitive patients. These altered modulations of the autonomic nervous system may contribute to the salt sensitivity of the blood pressure in patients with essential hypertension. PMID- 9222939 TI - Effects of chronic hormone replacement on the renin-angiotensin system in cynomolgus monkeys. AB - OBJECTIVE: To characterize the effects of estrogen, estrogen combined with progestin, and no treatment in ovariectomized cynomolgus monkeys during long-term reproductive hormone replacement. METHODS: Forty-five surgically postmenopausal cynomolgus monkeys fed a lipid-lowering diet were administered a conjugated equine estrogen (Premarin, 7.2 micrograms/day for the first 8 months, then 166 micrograms/day for the remaining 22 months), alone or in combination with 650 micrograms/day medroxyprogesterone acetate (Cycrin) for 30 months, or left with no hormone replacement therapy. Animals were anesthetized with ketamine pentobarbital, and samples were taken for measurements of plasma renin activity, angiotensin converting enzyme activity, and angiotensin peptides, angiotensin I (Ang I), angiotensin II (Ang II), and angiotensin-(1-7) [Ang-(1-7)]. RESULTS: Chronic replacement therapy with estrogen resulted in a significant elevation of the plasma renin activity [11.7 +/- 2.0 ng/ml per h control versus 22.8 +/- 4.6 ng/ml per h with estrogen (P < 0.05) versus 32.8 +/- 4.9 ng/ml per h with combination therapy (P < 0.01)], whereas estrogen or combination therapy caused a significant reduction in angiotensin converting enzyme activity [229 +/- 8 nmol/ml per min control versus 189 +/- 10 nmol/ml per min with estrogen (P < 0.05) versus 196 +/- 11 nmol/ml per min with combination therapy (P < 0.05)]. Both of these changes in angiotensin processing enzymes observed during replacement therapy resulted in significant increases in plasma Ang I levels [46.7 +/- 12.5 pg/ml control versus 175.5 +/- 65.9 pg/ml with estrogen (P < 0.05) and 561.7 +/- 373.6 pg/ml with combination therapy (P < 0.05)]. Plasma Ang II and Ang-(1-7) levels were not significantly changed. The mean blood pressure did not change with either treatment. CONCLUSION: These studies reveal that, although chronic estrogen replacement activates renin activity and Ang I, it causes a shift in the processing of angiotensin peptides such that the concurrent reduction in angiotensin converting enzyme activity leads to unchanged plasma Ang II levels. Thus, the potentially harmful effects of estrogen-induced hyperreninemia are balanced by its actions interfering with the formation of the vasoactive product Ang II. PMID- 9222941 TI - Longitudinal association of ambulatory pulse pressure with left ventricular mass and vascular hypertrophy in essential hypertension. AB - OBJECTIVE: To determine the longitudinal relationship between clinic and ambulatory blood pressures and subsequent left ventricular and carotid artery structure. DESIGN: A retrospective follow-up study. SETTING: A large district general hospital in Harrow, UK. PATIENTS: One hundred and forty patients who had been subjected to 24 h ambulatory intra-arterial blood pressure monitoring on the basis of their having an elevated clinic blood pressure were followed up randomly a mean of 9.4 +/- 3.4 years later. The ambulatory blood pressure parameters measured were the mean systolic, mean diastolic and mean pulse pressures. Follow up variables assessed included the clinic blood pressure, body mass index, total cholesterol, number of years of follow-up, left ventricular mass index, carotid intima-media thickness and carotid artery cross-sectional area. MAIN OUTCOME MEASURES: The left ventricular mass index, carotid intima-media thickness and carotid artery cross-sectional area. RESULTS: The mean pulse pressure and mean systolic blood pressure were correlated significantly with the left ventricular mass index (r = 0.46, P < 0.001 and r = 0.36, P < 0.001, respectively), carotid intima-media thickness (r = 0.45, P < 0.001 and r = 0.37, P < 0.001, respectively) and carotid artery cross-sectional area (r = 0.46, P < 0.001 and r = 0.41, P < 0.001, respectively). The mean pulse pressure was associated independently with all three outcome measures. In addition, the body mass index was an independent determinant of the left ventricular mass index, whereas the serum cholesterol level was associated independently with the carotid artery cross-sectional area; the number of years of follow-up was related independently to the left ventricular mass index and carotid intima-media thickness, but not to the cross-sectional area. CONCLUSIONS: These findings suggest that ambulatory blood pressure monitoring can play a role in guiding the choice of doses in drug therapy to limit potential target organ damage. PMID- 9222942 TI - Is insulin action a determinant of left ventricular relaxation in uncomplicated essential hypertension? AB - OBJECTIVE: To examine the relation of insulin action and left ventricular diastolic function in uncomplicated essential hypertension. METHODS: Doppler echocardiography and glucose clamping combined with indirect calorimetry were performed in 29, newly diagnosed, hypertensive men, free from cardiac and metabolic drugs. They were divided into two groups according to the clamp-derived whole-body glucose disposal level: 20 with insulin resistance (whole-body glucose disposal < 33 mumol/kg per min) and nine with normal insulin sensitivity. RESULTS: The two groups were comparable in age, body mass index, heart rate and blood pressure. No difference in diastolic function was found except for the isovolumic relaxation time, which was prolonged for patients with insulin resistance (P = 0.02). For the population as a whole, the relaxation time had univariate relations with the left ventricular mass index (r = 0.57, P < 0.001), whole-body glucose disposal (r = -0.56, P < 0.001) and non-oxidative glucose metabolism (r = -0.54, P = 0.002). In a multivariate model including age, body mass index, heart rate, diastolic blood pressure, left ventricular mass index and whole-body glucose disposal as potential determinants, only the left ventricular mass index (beta = 0.39, P = 0.02) and whole-body glucose disposal (beta = -0.38, P = 0.03) were independent predictors of the relaxation time (R2 = 0.43, P < 0.001). CONCLUSIONS: In uncomplicated essential hypertension the insulin resistance is a determinant of abnormalities in isovolumic relaxation, independently from the influence exerted by increased blood pressure levels, being overweight and left ventricular hypertrophy. PMID- 9222943 TI - Inhibition of nitric oxide in rats. Regulation of cardiovascular structure and expression of insulin-like growth factor I and its receptor messenger RNA. AB - OBJECTIVE: To determine whether 12 days' treatment with NG-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor, in spite of the increased arterial load, resulted in a growth-inhibitory response in the heart, aorta and skeletal muscle vascular bed, and whether the presence of L-NAME affected the expression of insulin-like growth factor-I and its receptor messenger RNA (mRNA). METHODS: Wistar rats were treated orally either with 100 mg/kg L-NAME or with tap water. On days 2, 4, 7 and 12 after initiation of treatment, the systolic blood pressure/mean arterial blood pressure and heart rate were measured, rats were killed and their heart and aorta were excised. Insulin-like growth factor-I and its receptor mRNA were quantitated by solution hybridization assay. On day 12 resistance properties in the skeletal muscle vascular bed were measured by using an in-vivo constant-flow preparation. RESULTS: The blood pressure in L-NAME-treated rats was increased immediately after initiation of treatment and it continued to increase throughout the experimental period. No hypertrophy was noted in the heart. Moreover, a 21% (P < 0.05) decrease in the right: left ventricular weight ratio indicated that attenuation of growth of the right ventricle had occurred. Increased expression of insulin-like growth factor-I and its receptor mRNA was observed neither in the heart nor in the aorta. The skeletal muscle vascular bed showed a 26% increased resistance at maximal vasodilatation (P < 0.05), which was indicative of a reduced average lumen size. A lower than expected perfusion pressure at maximal vasoconstriction was observed (17% above control, P < 0.05), implicating only modest medial thickening. CONCLUSION: L-NAME hypertension caused a prompt increase in blood pressure, which led neither to left ventricular hypertrophy nor to the expected overexpression of left ventricular/aortic insulin-like growth factor-I mRNA and only to partial structural adaptation in the skeletal muscle vasculature. These findings suggest that augmented expression of insulin-like growth factor-I and its receptor could be mandatory for conveying an appropriate adaptive hypertrophic response, at least in the heart. PMID- 9222944 TI - Effects of carbidopa and of intravenous saline infusion into normal and hypertensive subjects on urinary free and conjugated dopamine. AB - OBJECTIVE: To investigate the possible role played by endogenous dopamine as a modulator of renal sodium (Na+) reabsorption after a combined Na+ and volume load. DESIGN: A randomized placebo-controlled study. METHODS: Ten healthy volunteers and four hypertensive patients were subjected to intravenous infusions of 21 0.9% saline (308 mmol Na+) administered from 1000 to 1300 h after oral administration of placebo or of carbidopa, a dopamine decarboxylase inhibitor. RESULTS: Studies on control subjects after placebo showed that natriuresis occurred during the 6 h after commencement of the saline infusion, with falls in plasma albumin concentration, plasma renin activity and plasma aldosterone concentration; in comparison with results of mock infusion (6 mmol Na+) there was no change in the urinary excretion of dopamine and noradrenaline (In their free or conjugated forms). There was, however, a marked surge in excretion of urinary conjugated dopamine and in the dopamine: noradrenaline ratio from 1300 to 1600 h, after either type of infusion. Administration of carbidopa before the saline infusion resulted in a marked decrease in excretion of urinary free dopamine, but had no effect on the surge in excretion of urinary conjugated dopamine. Saline infusion decreased proximal fractional Na+ reabsorption. Administration of carbidopa delayed but did not prevent this decrease. The effects of saline infusion and of carbidopa on the urinary excretion of dopamine and noradrenaline from hypertensive patients were similar to those observed with the healthy volunteers. CONCLUSIONS: These findings indicate that volume expansion by intravenous saline infusion has no appreciable effect on the urinary free dopamine excretion from normal or hypertensive humans; with any apparent increase, it is important to exclude the possibility of conversion of conjugates to free dopamine in vitro. Furthermore, that carbidopa administration did not inhibit the afternoon surge of conjugated dopamine suggests that administration of carbidopa is deficient as a tool to investigate the functional role of the renal dopamine system. PMID- 9222945 TI - Changes in reactivity towards 5-hydroxytryptamine in the renal vasculature of the diabetic spontaneously hypertensive rat. AB - BACKGROUND: Diabetes and hypertension are both associated with an increased risk of renal disease. The combination of these diseases produces marked acceleration of the problems. OBJECTIVE: To examine the reactivity in the renal vasculature of diabetic hypertensive rats. METHODS: We investigated the reactivity towards 5 hydroxytryptamine (5-HT) in Krebs solution-perfused kidneys of diabetic normotensive Wistar-Kyoto (WKY) rats and spontaneously hypertensive rats (SHR). In addition, the interaction between the thromboxane A2 mimetic U46619 and 5-HT was examined. RESULTS: Discrete dose-response curves were obtained for the response to 5-HT (0.1-30 micrograms/g kidney) in Krebs solution-perfused kidneys of control and diabetic WKY rats and SHR. The following order of reactivity was determined: control SHR > diabetic SHR > control WKY rats = diabetic WKY rats. The thromboxane A2 mimetic U46619 (10 ng/ml) potentiated responses to 5-HT significantly in kidneys of diabetic WKY rats and control SHR, but not in kidneys from control WKY rats and diabetic SHR. CONCLUSIONS: The differential affected reactivity towards 5-HT kidneys from diabetic and hypertensive rats might be due to previously documented differences in receptor number. The marked effect of U46619 on the reactivity towards 5-HT in kidneys of diabetic and control rats indicates that this interaction might be important given the increased levels of thromboxane A2 reported to occur in these diseases. The reason for the lack of effect of thromboxane/prostaglandin receptor stimulation on responses to 5-HT in the combined model (i.e. diabetic hypertensive rats) needs to be determined. PMID- 9222946 TI - Antihypertensive action of non-natriuretic doses of furosemide in Dahl salt sensitive rats. AB - BACKGROUND: That non-natriuretic doses of loop diuretics exert an antihypertensive action has been suggested, but not confirmed, by simultaneous measurements of the arterial pressure and sodium balance during therapy. OBJECTIVE: To examine the relationship between changes in arterial pressure and changes in sodium balance during furosemide treatment. DESIGN: Twenty hypertensive Dahl salt-sensitive rats fed a 4% NaCl diet were allocated to four groups and administered the following treatments: placebo once a day intraperitoneally, continuous infusion of 4 mg/day furosemide intraperitoneally, 4 mg furosemide once a day intraperitoneally and 12 mg furosemide once every third day intraperitoneally. METHODS: The mean arterial pressure (MAP) was measured continuously with radiotelemetry and the sodium balance was measured with the rats in metabolic cages. RESULTS: Administration of furosemide as a bolus injection once a day (P < 0.01) or once every third day (P < 0.05) lowered the MAP significantly compared with placebo, whereas continuous infusion of furosemide had no significant effect on the MAP (P < 0.07). Fast Fourier transformation analysis detected an acute antihypertensive action related to the temporary diuretic and natriuretic responses during the period 0-6 h after intraperitoneal bolus injections of 4 and 12 mg furosemide. None of the treatment regimens produced 24 h sodium or potassium losses. At the end of the study, the total body water, extracellular fluid volume, total body sodium and potassium were similar for rats in all groups. CONCLUSIONS: Furosemide has an acute antihypertensive action in Dahl salt-sensitive rats fed a 4% NaCl diet that is related to renal sodium and volume losses whereas the long-term antihypertensive effect is independent of changes in extracellular fluid volume, total body water, sodium and potassium. PMID- 9222947 TI - Blood pressure effects of antihypertensive drugs and changes in lifestyle in a Brazilian hypertensive cohort. AB - BACKGROUND: The antihypertensive efficacy of drug therapy and of some nonpharmacologic recommendations has been demonstrated in controlled clinical trials, but not in a clinical setting. OBJECTIVE: To assess the antihypertensive effectiveness of drug therapy and of three nonpharmacologic recommendations (loss of weight, salt-intake restriction, and physical exercise). DESIGN: A prospectively planned cohort study. SETTING: A hospital-based hypertensive outpatient clinic. PATIENTS: We studied 637 patients (65.5% women) with systolic blood pressures above 140 mmHg or diastolic blood pressures above 90 mmHg, corresponding to 76% of 839 patients who were administered a prescription for hypertension and who returned for the first follow-up visit 3.5 months later on average. METHODS: The nonpharmacologic prescription consisted of salt-intake restriction for all, weight reduction for overweight patients, and practice of aerobic physical exercise for those for whom it was not contraindicated; 60% of the patients were treated with drugs according to standard recommendations. Patients treated with drugs were compared with untreated subjects; for the nonpharmacologic interventions, the groups were compared according to their reported compliances with the recommendations (at least some compliance versus none). The main outcome measures were variations in systolic and diastolic blood pressures between the baseline evaluation and the first follow-up visit and an improvement in prognosis, represented by a favorable change in the classification of the blood pressure (according to Joint National Committee V criteria). RESULTS: The cohort constituted predominantly low-income, middle-aged, overweight white women, with low-to-moderate hypertension of long duration. The group treated with drugs exhibited the greatest reduction in blood pressure, with clinical significance even discounting the losses in follow-up; the group of patients who reported compliance with the low-energy-intake diet also showed a consistent antihypertensive effect, which was still detectable on the occasion of the third follow-up visit 9 months after the first prescription; reported compliance with a low-sodium diet and practice of physical exercise were not associated with a reduction in blood pressure; among a subset of the patients, reported compliance with the salt-intake-restricted diet did not reduce the amount of sodium to the theoretical antihypertensive threshold. It was not possible to determine whether the lack of an antihypertensive effect of physical exercise for this cohort was secondary to a misreport of the extent of compliance or to an absence of effect of the intensity of training prescribed. The effects of drug therapy and compliance with a low-energy-intake diet were shown to be independent of other interventions or confounders. CONCLUSION: The antihypertensive effect of drugs demonstrated in well-controlled clinical trials is achievable in clinical practice. The recommendation to lose weight was the only nonpharmacologic intervention with a detectable antihypertensive effect in this cohort. The absence of effect of a low-sodium diet is probably secondary to the insufficient reduction in the amount of salt consumed. The lack of an antihypertensive effect of physical exercise could reflect either a misreported compliance or an absence of effect of the intensity of training recommended in this study. PMID- 9222948 TI - Effect of angiotensin converting enzyme inhibition after acute myocardial infarction in patients with arterial hypertension. TRACE Study Group, Trandolapril Cardiac Event. AB - OBJECTIVE: To evaluate the influence of a history of arterial hypertension on the efficacy of the angiotensin converting enzyme (ACE) inhibitor trandolapril in patients with acute myocardial infarction (AMI) and left ventricular dysfunction. METHODS: A retrospective analysis of data from the Trandolapril Cardiac Event (TRACE) study. The TRACE study was a randomized, double-blind, placebo-controlled study in which patients with an enzyme-verified AMI and ejection fraction < or = 35% were assigned randomly to be administered oral trandolapril or placebo 3-7 days after the infarction. Of 1749 patients who entered the study, 400 (23%) had a history of arterial hypertension. The mean follow-up time was 26 months. MAIN OUTCOME MEASURES: Mortality from any cause. Secondary endpoints were sudden death, cardiovascular mortality, reinfarction and development of severe heart failure. RESULTS: Of the patients in the hypertensive group, 173 (43%) died during follow-up, versus 500 (37%) in the normotensive group. Treatment with trandolapril resulted in a relative risk of death from any cause for the hypertensive group of 0.59 (96% confidence interval 0.44-0.80), versus 0.85 (0.72 1.02) for normotensive patients. In a multivariate analysis, treatment with trandolapril was associated with a reduction in mortality among patients with a history of hypertension (P = 0.03). CONCLUSION: In this retrospective analysis, ACE inhibition after AMI complicated with left ventricular dysfunction was of greater benefit to patients with a history of arterial hypertension. ACE inhibition might be of particular importance in this group of patients but further studies to establish the clinical impact are necessary. PMID- 9222949 TI - Biomechanical evaluation of an artificial anal sphincter prosthesis. AB - An artificial anal sphincter was designed to reproduce the normal physiology of the ano-rectum by flattening and angulating the bowel without causing crenation. It was shown in the laboratory that by eliminating crenation, occlusion pressure was evenly distributed without localized high pressure points under the device. Further, the design facilitates the creation of angulation of the bowel in a relatively large area of distribution of the occlusion pressure, allowing the device to be set at lower operating occlusion pressures than that required for circular designs yet still achieve continence. Each of the components of the device was studied separately and the whole system was assessed in an acute and survival animal model. PMID- 9222950 TI - A solid-state ambulatory physical activity monitor and its application to measuring daily activity of the elderly. AB - An ambulatory acceleration monitor was developed and used to evaluate physical activity in the elderly. The ECG and accelerations at the wrist, waist, and ankle were recorded, and the heart rate and frequency and amplitude of accelerations per unit time were evaluated in both working and retired elderly women performing normal daily activities. The heart rate closely paralleled wrist acceleration. The behaviour patterns of the subjects could be differentiated by time interval signals and histogram distributions. This system appears to be an effective method of understanding and describing the characteristic activities of the elderly in a quantitative and objective fashion. PMID- 9222951 TI - Design and development of a new cryosurgical instrument utilizing the Peltier thermoelectric effect. AB - Warts and some other dermatological conditions may be treated by the application of intense cold. This freezing has to be so severe as to form ice crystals which will rupture the cell membranes. The initial part of this project was to investigate the feasibility of using Peltier thermoelectric coolers (TECs) to cool down a suitable hand-held treatment tip to a temperature of approximately 50 degrees C. The results of these initial experiments showed how this could be accomplished and a prototype cryosurgical instrument, suitable for clinical trials, was designed and constructed. This new design is freestanding, self contained and is operated from a standard 230 V mains supply. Unlike existing systems it does not use any disposable gases or liquids. A cryoprobe of this new design would allow reliable cryosurgery to be performed in a GP's treatment room where supplies of liquid nitrogen, nitrous oxide or carbon dioxide are not readily available. The design also has a built-in thermometer to measure the treatment tip temperature thus ensuring consistency in treatment. Clinical trials are being conducted at a number of GP practices to evaluate this new design. PMID- 9222952 TI - Individual assessment of antihypertensive response by self-starting cumulative sums. AB - A self-interpreted control chart, on an individualized basis, assesses the effect of a switch from beta-blockers to an angiotensin-converting enzyme (ACE) inhibitor in a patient with occasional blood pressure (BP) excess. In dense and long data series, the BP and heart rate (HR) of this patient respond to the change in treatment by the test criterion of a self-starting Cumulative Sum (cusum), which reaches values outside a decision interval with a lowering of BP and an increase in HR and vice versa, at least for BP, after treatment cessation. Thereafter, minimal sampling requirements are sought in the same data by applying the same control chart approach to decimated data. Skeleton sampling schemes in a system of chronobiologic self-analysis and interpretation of manually recorded data obtained at strategically placed times (established on the basis of data decimations) could complement control charts that are used on a home computer or preferably would be built into the output of ambulatory monitors used at the outset as a minimum and routinely as an optimum. PMID- 9222953 TI - Assessing clot lysis strategies using a simplified mathematical model. AB - This paper attempts to describe lysis of a clot by infusion of lytic agent using a simple geometrical approach, in which the rate of clot lysis is assumed proportional to the exposed surface of the clot and the concentration of lytic agent. Six simple realizations (a)-(f) of this basic model are developed which account for the dependence of clot lysis time on five different clot geometries. In all six cases the clot is initially described as a uniform cylinder which totally occludes a vessel. In model (a) lysis proceeds as an advancing front at the proximal face of the clot. In model (b) lysis proceeds radially outwards from the central axis of the vessel while in model (c) lysis occurs radially inwards from the surface adjacent the wall, of the cylinder. In models (d) and (e) it is assumed that the clot breaks into a uniform spherical and cylindrical fragments, respectively, while model (f) uses the spherical fragment model combined with a lytic agent concentration which decreases with time. The validity of the models was assessed using previously published data from 76 patients in whom lysis time and clot size were recorded. Least squares linear regression analysis based on the six model equations yielded highly significant correlation coefficients r2 of 0.457, 0.412, 0.412, 0.495, 0.469, 0.663 for models (a)-(f), respectively. The results suggest that when a constant lytic agent concentration is assumed, no single geometry accounts for significantly more variation than any other, but that a combination of varying lytic agent concentration and clot geometry significantly influences clot lysis time and accounts for much of the observed variation. PMID- 9222954 TI - Construction of age-related reference limits for 24-h blood pressure pattern. AB - Blood pressure (BP) is recursively variable during the day-night cycle because of a physiological circadian rhythm. The aim of this study is, therefore, to show how to construct the population reference limits (desms) for BP in its time varying 24-h pattern, starting from a small sample, in order to facilitate their use in a local context. The sample for standardization comprised 427 clinically healthy subjects (211 males and 216 females), ranging in age from 16 to 100 years, attending their routine activities. The procedure begins with the statistical biometry related to the sample, and proceeds with the computation of the BP desms related to (1) the time-qualified discrete values; (2) the parameters of circadian rhythm; (3) the daily pressure load. The pertaining rules are explained step by stop, allowing each one to prepare the proper local desms for BP 24-h pattern. These standards may be useful for validating the individual BP monitoring according to the upper limits of the circadian physiological variability in the diagnostic procedure for identifying hypertensive subjects. PMID- 9222955 TI - Effect of vibrations with two different amplitudes on the ECG. AB - The ECG response of a human body exposed to vertical vibrations was investigated. The per cent normalized difference (PND) between the R-wave amplitude before and after vibrations was monitored. Results obtained from a representative subject show an oscillatory decay of the PND behaviour with time. This behaviour can be modelled as a linear second order damped system. The natural frequency, damping ratio and the recovery time of the cardiac system can be measured. PMID- 9222956 TI - Interference to medical equipment from mobile phones. AB - Cellular mobile phones may interfere with hospital equipment. We irradiated five representative pieces of equipment using simulated phone signals and frequencies. Two (an oximeter and a syringe pump) were immune to electric fields of up to 40 V m-1. The most susceptible was a physiological monitor which showed effects at 10 V m-1. None of the equipment was affected by fields that could be produced at over one metre from a 2 W mobile phone. PMID- 9222957 TI - A method of testing the compliance of 15 and 22 mm conical connectors on heat and moisture exchangers to ISO 5356-1:1987. AB - Heat and moisture exchangers (HMEs) are used to warm and humidify the gas delivered to patients during anaesthesia and in intensive care. Typically, HMEs are connected between the tracheal tube or catheter mount and the breathing system. This point of connection has been identified as having a high risk of disconnection. To reduce the possibility of disconnection, the International Standard for HMEs specifies that the connectors on an HME shall comply with ISO 5356-1: 1987, the International Standard for cones and sockets. A simple method is described to test whether or not the connectors comply with this standard. This method is used when evaluating HMEs for the Medical Devices Agency (MDA). Results are quoted from the MDA series Evaluation. The majority of connectors tested were found to comply with the standard. The test method is also suitable for testing conical connectors on other devices. PMID- 9222958 TI - A new X linked recessive syndrome of mental retardation and mild dysmorphism maps to Xq28. AB - Efforts to understand the genetic basis of mental retardation are greatly assisted by the identification of families with multiple relatives with mental retardation that clinical geneticists encounter in the routine practice of their profession. Here we describe a linkage study of a four generation family in which X linked recessive mental retardation (XLMR) is associated with minor dysmorphism and premature death of the affected males. Microsatellite based polymorphic loci evenly spaced over the entire X chromosome were used initially to detect linkage to Xq28. Further analysis identified a haplotype of Xq28 markers bounded proximally by locus DXS1113 and distally by DXS1108 that cosegregated with XLMR in this family. Two point lod scores > 3.0 provided strong evidence that the gene locus responsible for XLMR in this family is within this 7 Mb region of Xq28. The minor anomalies noted in some affected males were not distinctive enough to suggest a unique syndrome. None of our patients had features of the Waisman Laxova syndrome or the PPM-X syndrome. The possibility of allelism with any of the five other non-specific XLMR syndromes (MRX3, MRX16, MRX25, MRX28, and MRX41) mapped to Xq28 could not be excluded. While the recognition of a gene responsible for this disorder needs much additional work, multiple female relatives at risk in this family benefit immediately from knowing their genotype and heterozygotes will have the opportunity to undergo prenatal diagnosis. PMID- 9222959 TI - Fried syndrome is a distinct X linked mental retardation syndrome mapping to Xp22. AB - In 1972, Fried described a large Scottish family affected by X linked mental retardation (XLMR), hydrocephalus, and mild facial dysmorphism. The phenotype has considerable similarity to the MASA syndrome, which results from mutations of the L1CAM gene in Xq28, and this family has since been assumed to be an example of this condition. We have reinvestigated the family for linkage to X chromosome markers, and obtained additional clinical information on surviving affected subjects. The phenotype in these patients has evolved into a distinctive syndrome, with severe mental retardation (MR), spastic diplegia, ventricular dilatation, and calcification of the basal ganglia. Linkage to Xq28 markers has been excluded, suggesting that Fried syndrome is not allelic with MASA syndrome. Two point and multipoint linkage analysis indicates that the gene for this condition lies within the interval KAL-DXS989 in Xp22. We propose the designation Fried syndrome to emphasise the disorder's distinctive phenotype. PMID- 9222960 TI - Congenital renal tubular dysplasia and skull ossification defects similar to teratogenic effects of angiotensin converting enzyme (ACE) inhibitors. AB - An apparently autosomal recessive syndrome of congenital renal tubular dysplasia and skull ossification defects is described in five infants from two separate, consanguineous, Pakistani Muslim kindreds. The clinical, pathological, and radiological features are similar to the phenotype associated with fetal exposure to angiotensin converting enzyme (ACE) inhibitors: intrauterine growth retardation, skull ossification defects, and fetal/ neonatal anuric renal failure associated with renal tubular dysplasia. There was no fetal exposure to ACE inhibitors in the affected infants. Phenotypic similarities between these familial cases and those associated with ACE inhibition suggest an abnormality of the "renin-angiotensin-aldosterone" system (RAS). It is postulated that the molecular pathology in this uncommon autosomal recessive proximal renal tubular dysgenesis could be related to mutations of the gene systems governing the RAS. PMID- 9222961 TI - Genetic heterogeneity of primary open angle glaucoma and ocular hypertension: linkage to GLC1A associated with an increased risk of severe glaucomatous optic neuropathy. AB - The GLC1A locus for autosomal dominant juvenile and middle age onset primary open angle glaucoma (OAG) has been mapped to chromosome 1q21-q31. OAG, however, is a heterogeneous disease. We tested linkage of OAG and ocular hypertension (OHT), a major risk factor for OAG, to GLC1A in eight French families with multiple cases of juvenile and middle age onset OAG. There was strong evidence of genetic heterogeneity, four families being linked to GLC1A and two or three others being unlinked, depending on whether the complete OAG phenotype was analysed alone or jointly with OHT. Peak intraocular pressure (IOP) did not differ significantly between the two groups of families, while linkage to GLC1A conferred a highly increased risk of developing OAG and of having severe glaucomatous optic neuropathy. Testing linkage of familial OAG to GLC1A may therefore have prognostic value too. PMID- 9222962 TI - The effects of genotype and infant weight on adult plasma levels of fibrinogen, factor VII, and LDL cholesterol are additive. AB - High circulating levels of cholesterol, particularly low density lipoprotein (LDL) cholesterol and the clotting factors fibrinogen and factor VII, are associated with increased risk of myocardial infarction. Variations in the plasma levels of these factors are determined in part by polymorphisms in the genes concerned and also by weight at 1 year (infant weight). We have looked at the possibility of interactions between these genetic factors and infant weight in a sample of 290 men and 192 women from Hertfordshire using the beta-fibrinogen G/A 455, factor VII R353Q, and ApoE polymorphisms. The rare allele frequencies of the three polymorphisms were 0.19 for beta-fibrinogen, 0.10 for factor VII, and 0.07 and 0.13 for the 2 and 4 alleles of ApoE, and these frequencies were not different in subjects of different infant weight. In this sample, the polymorphisms showed the expected effects on plasma levels of fibrinogen, factor VII, and LDL cholesterol. The A-455 allele was associated with higher fibrinogen levels but the effect was only statistically significant in women (p = 0.003). The R353 allele was associated with higher factor VII activity in both men and women (p < 0.0001 for both). The ApoE2 allele was associated with lower levels of LDL cholesterol (p = 0.03 in men, p = 0.006 in women), while the ApoE4 allele was associated with higher levels (p < 0.001 in men, not significant in women). In this sample of men and women the effect of low infant weight was only associated with significant effects on fibrinogen and LDL cholesterol in the group of men (p = 0.005 and p = 0.008 respectively). Compared with the E3E3 subjects, the LDL lowering effect of the E2 allele and the raising effect of the E4 allele was greater in those with low infant weight compared with those with high infant weight (low v high infant weight for E2: 12.7% v 9.4%; for E4 12.7% v 8.5%). Although in this sample the interactive effect did not reach statistical significance, the additive effect of ApoE genotype and low infant weight on determining plasma LDL cholesterol levels, if confirmed, may be of relevance in determining a person's future risk of atherosclerosis. PMID- 9222963 TI - Progressive pseudorheumatoid dysplasia: report of a family and review. AB - Progressive pseudorheumatoid dysplasia is an inherited skeletal dysplasia with radiographic changes notably in the spine, similar to spondyloepiphyseal dysplasia tarda. There is also articular cartilage involvement which gives it some clinical resemblance to rheumatoid arthritis. We report here on six subjects from one inbred family from Jordan. Based on previously published reports and this one, we review the clinical and radiological features and discuss the genetics and differential diagnosis of the disorder. We suggest the addition of the word "spondyloepiphyseal" to the name adopted by the International Working Group on Constitutional Diseases of Bone, to become "progressive pseudorheumatoid spondyloepiphyseal dysplasia". We also speculate on candidate genes for this disorder. PMID- 9222964 TI - Intelligence indices in people with a high/low risk for developing Huntington's disease. AB - Intelligence in 20 presymptomatic subjects with an increased risk (> 95%) for carrying the gene for Huntington's disease (HD) was studied in a prospective, case-control, single blind study. No significant differences between the groups were detected for intelligence indices and subtest scores (Wechsler Adult Intelligence Scale). The high level of the performance IQ and the significant discrepancy between performance IQ and verbal IQ found in both the high risk and the low risk groups contrasted with our expectations based on anamnestic information, general clinical opinion, and the results of previously conducted studies. We propose that psychosocial circumstances could explain the test results and discuss the consequences of our findings for clinical genetics practice. PMID- 9222965 TI - Pitt-Rogers-Danks syndrome and Wolf-Hirschhorn syndrome are caused by a deletion in the same region on chromosome 4p 16.3. AB - Recently, a deletion of chromosome 4pter was found in three patients with Pitt Rogers-Danks syndrome. We investigated two of these patients, by means of DNA and FISH studies, together with two additional patients with Pitt-Rogers-Danks syndrome, to determine the critical region of the deletion in these patients and to compare this with the critical region in Wolf-Hirschhorn syndrome. All four patients showed terminal deletions of chromosome 4p of different sizes. One of them appeared to have an unbalanced karyotype caused by a cryptic translocation t(4;8) in the mother, resulting in a deletion of chromosome 4pter and a duplication of chromosome 8pter. The localisation of the Wolf-Hirschhorn critical region has been confined to approximately 1 Mb between D4S43 and D4S115. Our study shows that the deletions in four patients with the Pitt-Rogers-Danks syndrome overlap the Wolf-Hirschhorn critical region and extend beyond this in both directions. This study, combined with the fact that our third patient, who was previously described as a Pitt-Rogers-Danks patient, but who now more closely resembles a Wolf-Hirschhorn patient, makes it likely that Pitt-Rogers-Danks and Wolf-Hirschhorn syndromes are different clinical phenotypes resulting from a deletion in the same microscopic region on chromosome 4p16. PMID- 9222966 TI - Genetic linkage study of familial Mediterranean fever (FMF) to 16p13.3 and evidence for genetic heterogeneity in the Turkish population. AB - Familial Mediterranean fever (FMF) is an autosomal recessive condition that is almost entirely restricted to the non-Askhenazi Jews, Arabs, Armenians, and Turks. Genetic linkage study of a large group of non-Turkish families has previously mapped the FMF locus to the 16p13.3 region and shown that this locus resides 0.305 cM distal to D16S246. Furthermore, allelic association has also been shown with D16S3070 (75%) and D16S3275 (66%). However, no genetic heterogeneity has been described for any of the three major reported groups of FMF families. Here, we describe the genetic linkage relationship of the fourth major group of Turkish families and report the first evidence for genetic heterogeneity of this condition. Two point linkage analysis and haplotype inspection of 15 DNA markers from the reported region of the FMF locus identified tight linkage in a group of six Turkish FMF families. A maximum lod score of 9.115 at theta = 0.00 was observed for D16S3024. Nine other DNA markers provided similar evidence of linkage with lod score values of above 5.21. However, two other FMF families were completely unlinked to this region of chromosome 16. Haplotype construction of DNA markers in five consanguineous linked families showed that a segment of homozygosity has been conserved for D16S3070 and D16S2617. No other DNA markers showed any such conservation. Therefore, we suggested that these two markers reside in close proximity to the FMF locus. Furthermore, we observed 80% allelic association with D16S2617 but no association with D16S3070 or any other DNA markers from the FMF critical region. In summary, we conclude that our Turkish families are also linked to the reported FMF locus at 16p13.3, there is a genetic heterogeneity for this condition at least in our group of Turkish families, and D16S2617 is in linkage disequilibrium in the Turkish FMF families. Combination of this study with previously published observations suggests that the FMF locus resides between D16S246 and D16S3070/D16S2617 and within a region of about 250-300 kb. PMID- 9222967 TI - Four frameshift mutations in neurofibromatosis type 1 caused by small insertions. AB - We have been using heteroduplex analysis to assay individual exons within the NF1 gene in an effort to identify disease causing constitutional mutations in neurofibromatosis type 1 patients. Here we report the identification and characterisation of four insertional NF1 frameshift mutations in an analysis of exons 28-39 in a set of 78 patients. These include three 1 base pair insertions and one 2 base pair insertion. Three of these mutations can be attributed to replication slippage errors, while the mechanism behind the fourth may be related to formation of secondary structure during replication. It may be of significance that a majority of the previously reported small insertions in NF1 also lie within exons 28-39. PMID- 9222968 TI - Multiple lentigines syndrome (LEOPARD syndrome or progressive cardiomyopathic lentiginosis). AB - The multiple lentigines syndrome is an autosomal dominant condition which has many similarities to Noonan syndrome, except in the most striking feature from which its name is derived. The less neutral but very apt mnemonic, LEOPARD syndrome, was first used by Gorlin et al to whom the major debt in the definition of this syndrome lies, that is, Lentigines, ECG abnormalities, Ocular hypertelorism/Obstructive cardiomyopathy, Pulmonary valve stenosis, Abnormalities of genitalia in males, Retardation of growth, and Deafness. Not previously included in the mnemonic is cardiomyopathy which is an important feature because it is associated with significant mortality. PMID- 9222969 TI - A family with a milder form of adult dominant polycystic kidney disease not linked to the PKD1 (16p) or PKD2 (4q) genes. AB - Autosomal dominant polycystic kidney disease (ADPKD) is a genetically heterogeneous disease. Most families show positive linkage to polymorphic markers around the PKD1 (16p13.3) or PKD2 (4q21-23) loci. The PKD1 and PKD2 genes have been cloned and mutations defined in a number of patients. Several clinical studies have described a milder phenotype for PKD2 patients. More recently, evidence for a third genetic locus has been found in one Portuguese, one French Canadian, and one Italian family. We identified a Spanish family with negative linkage to the PKD1 and the PKD2 loci. This family showed a very mild clinical phenotype compared to the other forms of ADPKD, including the non-PKD1/non-PKD2 families previously described. PMID- 9222970 TI - Identification of a recombination event narrowing the Lafora disease gene region. AB - Patients affected with progressive myoclonus epilepsy of the Lafora type present during late adolescence with a characteristic EEG pattern and Lafora bodies seen on skin biopsy. The critical region for the Lafora gene has been localised to chromosome 6q24 flanked by the dinucleotide repeat markers D6S292 and D6S420. This study for linkage of markers from the candidate gene region was performed in a previously unpublished family affected with Lafora disease. EEG and skin biopsy evaluation for Lafora bodies were performed on five of eight family members followed for seizure activity. Haplotype and linkage analysis of DNA from five family members were carried out using the nine dinucleotide repeat markers reported in the common region of homozygosity by Serratosa et al in 1995. The present study of an additional family affected by Lafora disease has narrowed the 17 cM critical region for the Lafora disease gene on chromosome 6q24 to a 4 cM region flanked by markers D6S308 and D6S311. PMID- 9222971 TI - The 12 base pair duplication/insertion alteration could be a regulatory mutation. AB - A wide array of mutations now numbering more than 200 have been identified in the BRCA1 gene, one of the two breast cancer susceptibility genes identified so far. In addition, there have been several variants described but it is not known if they really represent functionally significant mutations of the BRCA1 gene. We report evidence to show that the duplication/ insertion of 12 base pairs in intron 20 could have a real effect on expression of the BRCA1 gene, although it was also present in 1% of our control population. PMID- 9222972 TI - Phosphoserine phosphatase deficiency in a patient with Williams syndrome. AB - Decreased serine levels were found in plasma and cerebrospinal fluid (CSF) of a boy with pre- and postnatal growth retardation, moderate psychomotor retardation, and facial dysmorphism suggestive of Williams syndrome. Fluorescence in situ hybridisation with an elastin gene probe indicated the presence of a submicroscopic 7q11.23 deletion, confirming this diagnosis. Further investigation showed that the phosphoserine phosphatase (EC 3.1.3.3.) activity in lymphoblasts and fibroblasts amounted to about 25% of normal values. Oral serine normalised the plasma and CSF levels of this amino acid and seemed to have some clinical effect. These data suggest that the elastin gene and the phosphoserine phosphatase gene might be closely linked. This seems to be the first report of phosphoserine phosphatase deficiency. PMID- 9222973 TI - A case of apparent trisomy 21 without the Down's syndrome phenotype. AB - We describe a case of apparent trisomy 21 that does not fulfill the criteria for the clinical diagnosis of Down's syndrome (DS). Our patient was subjected to karyotype analysis and found to have full, non-mosaic trisomy 21 in both blood lymphocytes and skin fibroblasts, while examination of the term placenta, which was performed earlier in the course of a different study, had shown mosaicism (73%) for trisomy 21. FISH analysis showed no obvious rearrangement of the DS chromosomal region in any of the chromosomes 21. Molecular analysis using polymorphic markers on chromosome 21 verified the existence of trisomy for the entire long arm of the chromosome and showed that the origin of the extra chromosome was maternal and was probably the result of a mitotic error. In contrast with the above, the clinical evaluation using the Jackson checklist of 25 signs failed to establish the diagnosis of DS. We believe that our patient might present mosaicism in other tissues that are not available for analysis and can be regarded as an extreme example in the continuous spectrum of karyotype phenotype associations in mosaic cases. PMID- 9222974 TI - Congenital diaphragmatic hernia with probable autosomal recessive inheritance in an extended consanguineous Pakistani pedigree. AB - We report four cases of congenital diaphragmatic hernia occurring in two generations of a consanguineous Pakistani family. The affected subjects resembled no recognised genetic syndrome. This family provides further evidence for possible autosomal recessive inheritance of congenital diaphragmatic hernia in some cases. PMID- 9222975 TI - A case of Lenz microphthalmia syndrome. AB - Lenz microphthalmia syndrome was first described by Lenz et al in 1955. The cardinal features of the syndrome are microphthalmia or anophthalmos, narrow shoulders, other skeletal anomalies, and dental and urogenital malformations. Here we present a case of Lenz microphthalmia syndrome who shows the typical characteristics and, additionally, dysgenesis of the corpus callosum associated with dilatation of the lateral ventricles. The patient, a 13 year old male, was referred to our hospital by a dental hospital for genetic counselling. On physical examination, height, weight, and head circumference were below the 3rd centile and he had brachymicrocephaly, a preauricular tag, microphthalmia, missing teeth, narrow shoulders, long, proximally placed thumbs, hypospadias, cryptorchidism, and a normal IQ. Ophthalmological examination showed microcornea, sclerocornea, absence of the pupil, no vision in the left eye and decreased vision and a small pupil in the right eye in addition to his bilateral microphthalmia. Cranial MRI showed dilatation of the lateral ventricles and dysgenesis of the corpus callosum. PMID- 9222976 TI - The mitochondrial A3243G mutation presenting as severe cardiomyopathy. AB - A 6 year old Portuguese boy with dilated cardiomyopathy had abundant ragged red fibres in muscle (20% of total) and severe lactic acidosis. Molecular genetic analysis showed the A to G transition in the mitochondrial transfer RNALeu(UUR) gene at nt 3243 ("MELAS mutation"), which accounted for 88% and 68% of the total mtDNA in his muscle and blood, respectively. Molecular studies in blood from 16 maternal relatives identified lower percentages of the mutation only in the oligo symptomatic mother and brother. This case reinforces the notion that cardiomyopathy can be the presenting and predominant clinical expression of the A3243G mutation. PMID- 9222978 TI - Marfan syndrome. PMID- 9222977 TI - A report of a child with a deletion (9)(q34.3): a recognisable phenotype? AB - We report a case of a male infant who presented with congenital anomalies and was found to have a de novo deletion in the terminal region of the long arm of chromosome 9. He died at the age of 17 weeks of cardiorespiratory failure owing to RSV positive bronchiolitis. A review of previously published reports documented one previous report of a patient with a deletion of (9)(q34.3) and multiple congenital anomalies. Comparison with the previously reported case suggests that the phenotype observed constitutes a clinically recognisable pattern of malformations. PMID- 9222979 TI - Protein design as a challenge for peptide chemists. AB - All efforts to turn the ultimate goal in protein de novo design into reality-the construction of new macromolecules with predetermined three-dimensional structure and well-defined functionality-failed because the mechanism of folding has still to be unravelled. In the present review, various attempts to apply synthetic tools for inducing native-like structural features in peptides in order to bypass the folding problem are described. Besides well-established methods for the nucleation and stabilization of secondary structures, e.g. alpha-helices, beta sheets and beta-turns, topological templates as 'built-in' folding devices have more recently become the key elements for the induction of protein-like folding units (template-assembled synthetic proteins, TASP). Progress in the synthetic strategy and structural characterization of this new type of macromolecules opens the way for the design of functional TASP molecules. PMID- 9222980 TI - N,O-bisFmoc derivatives of N-(2-hydroxy-4-methoxybenzyl)-amino acids: useful intermediates in peptide synthesis. AB - 2-Hydroxy-4-methoxybenzyl-amino acid residues inhibit interchain association in solid phase peptide synthesis. They are easily introduced through their N,O bisFmoc derivatives. Preparation of a range of these derivatives is described. PMID- 9222981 TI - Binary synthesis of multicomponent peptide mixtures by the portioning-mixing technique. AB - By introducing a new operation (non-coupling), our portioning-mixing method has become suitable for preparing binary peptide libraries. We demonstrate that all the expected components of a simple library are present in the mixture. The number of components in such libraries, the molar ratio of peptides as well as the possibilities of screening are discussed. PMID- 9222982 TI - Synthesis, characterization and biocompatibility of PEGA resins. AB - Three types of beaded polyethylene glycol polyacrylamide copolymers (PEGA) with a high content of polyethylene glycol (PEG) were synthesized by inverse suspension polymerization and characterized for peptide synthesis and with respect to their physical properties. Several peptides of high purity have been synthesized on the resin. The properties which were determined were loading of amino groups, swelling, bead size distribution, porosity, flexibility and compatibility with active biomolecules. A loading of 0.35 mmol/g has been obtained and the swelling was excellent in solvents of various polarities ranging from water to dichloromethane. The 13C-NMR T1-relaxation times of a resin containing a peptide were determined in DMSO-d6 and the resin was found to exhibit a behavior similar to the components in free solution. PMID- 9222983 TI - Synthesis and conformational studies of peptides containing TOAC, a spin-labelled C alpha, alpha-disubstituted glycine. AB - A variety of host L-alanine homo-peptides (to the pentamer) containing one or two spin-labelled TOAC (2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid) residues were synthesized by solution methods and fully characterized. The conformational features of the terminally blocked, doubly spin-labelled TOAC (Ala)2-TOAC-Ala-pentapeptide were examined in the crystal state by X-ray diffraction and in solution using a combination of techniques (Fourier transform infrared, circular dichroism, cyclic voltammetry and electron spin resonance) in comparison with singly labelled shorter peptides. The 3(10)-helical structure of the pentapeptide, promoted by the two C alpha, alpha-disubstituted glycines under favourable experimental conditions, allows an interaction to take place between the two nitroxide TOAC side chains spaced by one turn of the helix. Taken together, these results suggest that TOAC is an excellent probe for exploring bends and helices in doubly labelled peptides. PMID- 9222984 TI - Synthesis and iodination of human (phenylalanine 13, tyrosine 19) melanin concentrating hormone for radioreceptor assay. AB - An analogue of human melanin-concentrating hormone (MCH) suitable for radioiodination was designed in which Tyr13 and Val19 of the natural peptide were replaced by phenylalanyl and tyrosyl residues: [Phe13, Tyr19]-MCH. The peptide was synthesized by the continuous-flow solid-phase methodology using Fmoc strategy and polyhipe PA 500 and PEG-PS resins. The linear MCH peptides with either acetamidomethyl-protected or free cysteinyl residues were purified to homogeneity and cyclized by iodine oxidation, yielding the final product with the correct molecular weight of 2434.61. Radioiodination of the C-terminal tyrosine was carried out enzymatically using solid-phase bound glucose oxidase/lactoperoxidase, followed by purification on a reversed-phase mini-column and by high-pressure liquid chromatography. The resulting [125I]-[Phe13, Tyr19] MCH tracer was the first radiolabelled MCH peptide suitable for radioreceptor assay: saturation binding analysis using mouse G4F-7 melanoma cells demonstrated the presence of 1090 MCH receptors per cell. The dissociation constant (KD) was 1.18 x 10(-10) M, indicating high-affinity MCH receptors on these cells. MCH receptors were also found in other cell lines such as mouse B16-F1 and G4F and human RE melanoma cells as well as in PC12 and COS-7 cells. Competition binding analyses with a number of other peptides such as alpha-MSH, neuropeptide Y, substance P and pituitary adenylate cyclase activating peptide, demonstrated that the binding to the MCH receptor is specific. Atrial natriuretic factor was found to be a weak competitor of MCH, indicating topological similarities between MCH and ANF when interacting with MCH receptors. PMID- 9222986 TI - Multiple synthesis by the multipin method as a methodological tool. AB - The multipin method of peptide synthesis is demonstrated as a potent methodological tool, where large numbers of comparative studies can be performed concurrently. Two studies are presented. In each study, the test peptides were simultaneously synthesized, and the products examined by high throughput ion spray mass spectrometry and reverse-phase HPLC. In the first study, comprising 24 experiments, peptides 1 (AELFSTHYLAFKEDYSQ-NH2) and 2 (LKDFRVYFREGRDQLWKGPG-NH2) were prepared using Fmoc-Axx/BOP/HOBt/NMM [100 : 100 : 100 : 150 mM) and Fmoc AXX/HATU/HOAt/NMM (100 : 100 : 100 : 150 nM) with 60, 90 and 120 min coupling times. The two reagent combinations were found to give comparable results. The second study compared the N-terminal coupling of Fmoc-Asn-OH, Fmoc-Asn(Mbh)-OH, Fmoc-Asn(Mtt)-OH, Fmoc-Asn(Tmob)-OH and Fmoc-Asn(Trt)-OH in the synthesis of seven test peptides: 3, NVQAAIDYIG-cyclo(KP): 4. NTVQAAIDYIG-cyclo(KP): 5. NRVYVHPFNL: 6. NRVYVHPFHL: 7. NEAYVHDAPVRSLN: 8. NQLVVPSEGLYLIYSQVLFK; 9, NPNANPNANPNA. A total of 33 experiments were performed. Peptides 3 and 4 were selected to highlight the effect of steric bulk of each Asn derivative on coupling efficiency. Reagent efficiency, as measured by target peptide purity, was as follows: Fmoc-Asn(Tmob)-OH > Fmoc-Asn-OH > Fmoc-Asn(Mtt)-OH = Fmoc Asn(Trt)-OH > Fmoc-Asn(Mbh)-OH. PMID- 9222985 TI - An exploration of the effects of L- and D-tetrahydroisoquinoline-3-carboxylic acid substitutions at positions 2, 3 and 7 in cyclic and linear antagonists of vasopressin and oxytocin and at position 3 in arginine vasopressin. AB - We have investigated the effects of mono-substitutions with the conformationally restricted amino acid, 1,2,3,4 tetrahydroisoquinoline-3-carboxylic acid (Tic) at position 3 in arginine vasopressin (AVP), at positions 2, 3 and 7 in potent non selective cyclic AVP V2/V1a antagonists, in potent and selective cyclic and linear AVP V1a antagonists, in a potent and selective oxytocin antagonist and in a new potent linear oxytocin antagonist Phaa-D-Tyr(Me)-Ile-Val-Asn-Orn-Pro-Orn NH2 (10). We report here the solid-phase synthesis of peptide 10 together with the following Tic-substituted peptides: 1. [Tic3]AVP: 2. dICH2)5[D-TIc2]VAVP: 3, d(CH2)5[D-Tyr(Et)2Tic3]VAVP: 4, d(CH2)5[Tic2Ala-NH2(9)]AVP: 5. d(CH2)5[Tyr]Me)2.Tic3,Ala-NH2(9)]AVP: 6. d(CH2)5 [Tyr(Me)2,Tic7]AVP: 7, Phaa-D Tyr(Me)-Phe-Gln-Asn-Lys-Tic-Arg-NH2: 8, desGly-NH2,d[CH2]5[Tic2,Thr4]OVT: 9. desGly-NH2d(CH2)5[Tyr(Me)2Thr4, Tic7[OVT; 11, Phaa-D-Tic-Ile-Val-Asn-Orn-Pro-Orn NH2, using previously described methods. The protected precursors were synthesized by the solid-phase method, cleaved, purified and deblocked with sodium in liquid ammonia to give the free peptides 1-11 which were purified by methods previously described. Peptides 1-11 were examined for agonistic and antagonistic potency in oxytocic (in vitro, without Mg2+) and AVP antidiuretic (V2-receptor) and vasopressor (V1a-receptor) assays. Tic3 substitution in AVP led to drastic losses of V2, V1a and oxytocic agonistic activities in peptide 1, L- and D-Tic2 substitutions led to drastic losses of anti-V2/anti-V1a and anti oxytocic potencies in peptides 2, 4, 8 and 11 (peptide 2 retained substantial anti-oxytocic potency; pA2 = 7.25 +/- 0.025). Whereas Tic3 substitution in the selective V1a antagonist d(CH2)5[Tyr(Me)2,Ala-NH2(9)]AVP(C) led to a drastic reduction in anti-V1a potency (from anti-V1a pA2 8.75 to 6.37 for peptide 5, remarkably, Tic3 substitution in the V2/V1a antagonist d(CH2)5(D-Tyr(Et)2]VAVP(B) led to full retention of anti-V2 potency and a 95% reduction in anti-V1a potency. With an anti-V2 pA2 = 7.69 +/- 0.05 and anti-V1a pA2 = 6.95 +/- 0.03. d(CH2)5[D Tyr(Et)2, Tic3]VAVP exhibits a 13-fold gain in anti-V2/anti-V1a selectivity compared to (B). Tic7 substitutions are very well tolerated in peptides 6, 7 and 9 with excellent retention of the characteristic potencies of the parent peptides. The findings on the effects of Tic3 substitutions reported here may provide promising leads to the design of more selective and possibly orally active V2 antagonists for use as pharmacological tools and as therapeutic clinical agents for the treatment of the syndrome of the inappropriate secretion of antidiuretic hormone (SIADH). PMID- 9222987 TI - Antiovulatory antagonists of LHRH related to antide. AB - We report 104 analogues of the potent antiovulatory antagonist of LHRH, N-Ac-D Nal-D-Cpa-D-Pal-Ser-Lys(Nic)-D-Lys(Nic)-Leu-Ilys-Pro-D-Ala- NH2, Antide. We replaced the Nic group in Antide with other acyl substituents to modulate size, hydrophilicity or basicity of the molecule, we also replaced the Lys residues with shorter basic amino acids, and made cyclic 5/6 analogues as well as position 5 or 6 dimers. We substituted Ilys8 with other alkyl groups and acyl derivatives. When injected in 0.1% DMSO in water in a typical antiovulatory (AO) assay. Antide gives six rats ovulating out of eight (6/8) at 2 micrograms, 4/8 at 4 micrograms, and in the histamine release assay (HRA). ED50 is > 300 micrograms/ml; [Lys(N Isobutyl)8]Antide gave 2/8 at 2 micrograms/rat; [Lys (8-Qis)5]Antide gave 1/8 at 1 microgram, and 0/8 at 2 micrograms, and in the HRA ED50. 22 micrograms/ml; [D Lys(8-Qis)5]Antide gave 4/8 at 1 microgram and 0/8 at 2 micrograms, and in the HRA, ED50 was 27 micrograms/ml; [Lys(8-Qic)8] gave 5/8 at 1 microgram 1/8 at 2 micrograms/ [Lys(2-Pyc)6]Antide gave 3/8 at 1 microgram, and 0/8 at 2 micrograms, and in the HRA ED50 was 116 micrograms/ml; [D-Lys (2-Pyc)5]Antide gave 5/8 at 1 microgram and in the HRA, ED50 was 100- > 300 micrograms/ml; [Lys(2-Pyc)5.D-Lys(2 Pyc)6]Antide gave 2/8 at 1 microgram. The substitutions of the Nic groups of Antide at Lys5 or D-Lys6 with 8-Qis or with 2-Pyc groups seem to give highly potent antiovulatory antagonists of LHRH and constitute significant new leads to generate potent antiovulatory compounds endowed with moderate or low histamine release. PMID- 9222988 TI - Structural and immunological reactivity of the principal neutralizing determinant V3 of glycoprotein gp120 of HIV-1. AB - The variable domain V3 in the outer glycoprotein gp120 of HIV-1 is a highly important region with respect to immune response during the course of viral infection. Neutralizing antibodies are produced against this domain: in addition, it has been shown to be a functionally active epitope for T helper and cytotoxic T cells. The high degree of amino acid variability in individual HIV-isolates, however, limits the use of the V3-domain in approaches to vaccine development. In order to characterize the residues important for antibody interaction and binding to MHC class I proteins, we constructed a consensus sequence of the V3-domain with broad reactivity [1] and used synthetic peptides derived from this consensus sequence with individual residues altered to alanine. These peptides were used as antigens in ELISA tests to define the amino acids which are important for binding to human and rabbit/anti-peptide immunoglobulins. In addition, we used these alanine-derived peptides in interaction studies with human HLA-A2.1 and mouse H 2Dd by testing their capacity to stabilize the respective MHC class I protein complexes on the surface of mutant cell lines T2 and RMA-S transfected with Dd gene. The experimental tests allowed us to define individual residues involved in antibody and MHC-protein interaction, respectively. In a further approach, we used those results to design interaction models with HLA-A2.1 and H-2Dd. Therefore, a structural model for H-2Dd was built that exhibits an overall similar conformation to the parental crystal structure of HLA-A2.1. The resulting interaction models show V3-peptide bound in an extended beta-conformation with a bulge in its centre for both H-2Dd and HLA-A2.1 complexes. The N- and C-termini of V3 peptide reside in conserved pockets within both MHC-proteins. Anchoring residues could be determined that are crucial for the binding of the respective MHC class I haplotype. The cross-reactivity of V3-peptide in enhancing the expression of two different MHC class I molecules (H-2Dd and HLA-A2.1) is shown to be based on similar peptide binding that induces an almost identical peptide conformation. PMID- 9222989 TI - The use of N-urethane-protected N-carboxyanhydrides (UNCAs) in amino acid and peptide synthesis. AB - N-Urethane-protected N-carboxyanhydrides (UNCAs) are very reactive amino acid derivatives. They have been successfully used in peptide synthesis, in both solution and solid phase. We have demonstrated that UNCAs are interesting starting materials for the synthesis of various amino acid derivatives. Chemoselective reduction of UNCAs with sodium borohydride led the corresponding N protected beta amino alcohols. Reaction of UNCAs with Meldrum's acid, followed by cyclisation, yielded enantiomerically pure tetramic acid derivatives. Diastereoselective reduction of tetramic acid derivatives produced [4S,5S)-N alkoxycarbonyl-4-hydroxy-5-alkylpyrrolidin-2-ones derived from amino acids, which after hydrolysis yielded statine and statine analogues. Tetramic acid derivatives could also be obtained by reaction of UNCAs with benzyl ethyl malonate in the presence of sodium hydride to yield gamma-N-benzyloxycarbonylamino-beta oxodicarboxyl esters followed by hydrogenolytic deprotection and decarboxylation. UNCAs also reacted with phosphoranes to produce the ketophosphorane in excellent yields. Subsequent oxidation with oxone or with [bis(acetoxy)-iodo]-benzene produced vicinal tricarbonyl derivatives. These reactions usually proceeded smoothly and with high yields. PMID- 9222990 TI - A synthetic substrate assay for the gamma-secretase of the beta-A4 amyloid of Alzheimer's disease. AB - gamma-secretase, the endoprotease which releases the C-terminus of beta A4 amyloid peptide, cleaves within the hydrophobic transmembrane domain of the amyloid precursor protein. In order to obtain a substrate for gamma-secretase, a dodecapeptide which spans the cleavage site was synthesized, labelled with 125 iodine and conjugated to an agarose gel. A radiometric solid-phase assay was developed using this immobilized substrate. Peptide products were separated by reverse-phase HPLC and TLC to allow characterization of the cleavage site(s). PMID- 9222991 TI - Immunogenicity of dinitrocarboxyphenylated melittin: the influence of C-terminal chain shortening, N-terminal substitution and prolin insertion at positions 5 and 10. AB - Peptides derived from the bee-venom melittin were fitted with the haptenic group dinitrocarboxyphenyl (Dncp) and tested in out-bred guinea pigs for immunogenicity by measuring the IgG anti-Dncp antibody response by ELISA. Dncp-conjugates comprising virtually the entire melittin proved to be strong immunogens producing antibody responses comparable to those of proteins. Weak responses were obtained with considerably shortened sequences. Conjugates with N-terminal Dncp gave markedly reduced antibody responses compared to peptides with C-terminal Dncp. An N-terminal biotinyl substituent abolished the immune response whereas N-terminal lauryl and caprylyl had little effect. Insertion of L-proline into a hexadecapeptide conjugate abolishing the possibility of helix formation gave an immunogen to which individual animals clearly responded on a low level. Oligomerisation, but not the cytolytic activity of melittin peptides, may contribute to the immunogenicities observed. PMID- 9222992 TI - Proteinogenic amino acids labelled with 15N and/or 13C for application in peptide synthesis: a short review with a comprehensive list of published derivatives. AB - A review is given of the literature dealing with the most common protected derivatives of 15N- and/or 13C-labelled amino acids of interest in peptide synthesis. The list contains all such Boc-, Z- and Fmoc-amino acids as well as published methyl, ethyl, t-butyl and benzyl esters. PMID- 9222993 TI - Highly potent side chain-main chain cyclized dermorphin-deltorphin analogues: an integrated approach including synthesis, bioassays, NMR spectroscopy and molecular modelling. AB - Our continuing efforts to study structure-activity relationships of peptide opioids have resulted in the synthesis of a series of cyclic opioids related to dermorphins and deltorphins. The biological activities of the compounds have been determined and the conformational analyses carried out using 1H-NMR spectroscopy and molecular modelling. The three compounds in the series Tyr-c[D-Orn-Phe-Ala], Tyr-c[D-Lys-Phe-Ala], and Tyr-c[A2bu-Phe-Ala-Leu] are cyclized via a lactam bridge from the side-chain of the residue at the second position with the carboxyl terminus of each compound. The molecules incorporate 12-, 13- and 14 membered rings, respectively. They include a phenylalanine at the third position which is a distinguishing characteristic of dermorphins and deltorphins. The guinea pig ileum and mouse vas deferens assays show that the compounds are highly active at both mu- and delta-opioid receptors. The compounds are all highly effective antinociceptive agents as measured by the intrathecal rat hot plate test. Conformational analyses of the molecules indicate that they can adopt topochemical arrays required for bioactivity at both mu- and delta-receptors which explains their high activity in both guinea pig ileum and mouse vas deferens in vitro assays. The results support our models for mu- and delta receptor activity for constrained peptide opioids. PMID- 9222994 TI - Solid-phase synthesis of peptide nucleic acids. AB - Peptide nucleic acids (PNA) were synthesized by a modified Merrifield method using several improvements. Activation by O-[benzotriazol-1-yl]-1,1,3,3 tetramethyluronium hexafluorophosphate in combination with in situ neutralization of the resin allowed efficient coupling of all four Boc-protected PNA monomers within 30 min. HPLC analysis of the crude product obtained from a fully automated synthesis of the model PNA oligomer H-CGGACTAAGTCCATTGC-Gly-NH2, indicated an average yield per synthetic cycle of 97.1%. N1-benzyloxycarbonyl-N6(3) methylimidazole triflate substantially outperformed acetic anhydride as a capping reagent. The resin-bound PNAs were successfully cleaved by the 'low-high' trifluoromethanesulphonic acid procedure. PMID- 9222996 TI - Anchor-linked intermediates in peptide amide synthesis are caused by dimeric anchors on the solid supports. AB - Cleavage and kinetic studies have been carried out using commercially obtained H Tyr(tBu)-5-(4'-aminomethyl-3',5'-dimethoxyphenoxy)valeric acid-TentaGelS (H Tyr(tBu)-4-ADPV-TentaGelS) and H-Tyr (tBu)-4-ADPV-Ala-aminomethyl-resin (H Tyr(tBu)-4-ADPV-AM-resin) prepared from commercially available resin and loaded with commercially available Fmoc-4-ADPV-OH amide anchor. Cleavage with pure trifluoroacetic acid (TFA) gave the intermediate H-Tyr-4-ADPV-NH2, which was then degraded to H-Tyr-NH2, and cleavage with TFA/dichloromethane (1:9) yielded H-Tyr 4-ADPV-NH2 which could be isolated in preparative amounts. Cleavage reactions with 15N-labelled H-Ala-4-ADPV-(15N)-Gly-AM-resin yielded the intermediate H-Ala 4-ADPV-NH2, which contained no 15N as demonstrated by 1H-NMR. The analysis of the commercial Fmoc-4-ADPV-OH amide anchor showed the presence of Fmoc-4-ADPV-4-ADPV OH as an impurity in high amounts. This dimeric anchor molecule is the cause of formation of the anchor-linked peptide intermediate obtained during the cleavage from the resin. The particularly high acid-lability of the amide bond between the two ADPV moieties was utilized to synthesize sidechain and C-terminally 4-ADPV protected pentagastrin on a double-anchor resin, and to cleave it using 5% trifluoroacetic acid in dichloromethane. This method may offer a new way for the synthesis of protected peptide amides with improved solubility to be used in fragment condensation. PMID- 9222997 TI - Synthesis and properties of the first all-aza analogue of a biologically active peptide. AB - The synthesis of the first all-aza-amino acid analogue (2) of a peptidic renin inhibitor is described. The X-ray structural analysis and molecular modelling investigations of this novel compound reveal interesting conformational features which have a significant impact on its biological activity. In addition, insight into conformational features of azapeptides in general in comparison with the corresponding purely peptidic compounds is given. PMID- 9222995 TI - Synthetic S-(2,3-dihydroxypropyl)-cysteinyl peptides derived from the N-terminus of the cytochrome subunit of the photoreaction centre of Rhodopseudomonas viridis enhance murine splenocyte proliferation. AB - Various lipopeptides representing the N-terminal part of the cytochrome subunit of the photosynthetic reaction centre from the purple bacterium Rhodopseudomonas virdis were prepared by solid-phase peptide synthesis. These lipopeptides consisted of a S-[2,3-dihydroxypropyl]-cysteinyl (Dhc) residue N-terminally coupled to the nonapeptide FEPPPATTT. Different numbers of palmitoyl (Pam) chains were attached to Dhc via ester and/or amide bonds. The lipopeptide Dhc(Pam)2 FEPPPATTT containing two ester-bonded palmitoyl residues and a free N-terminus was a potent polyclonal activator of murine (BALB/c) spleen cells at subnanomolar concentrations. The lipopeptide Pam-Dhc(Pam)2-FEPPPATTT containing three palmitoyl residues, the two-chain lipopeptide Pam-Dhc(Pam)-FEPPPATTT containing one amide- and one ester-bonded palmitoyl residue, and the N-terminally elongated lipopeptide SLVAG-Dhc(Pam)2-FEPPPATTT were less active. The nonapeptide FEPPPATTT and the decapeptide Dhc-FEPPPATTT were only marginal splenocyte activators, even at concentrations as high as 1 microM. Thus, lipopeptide Dhc(Pam)2-FEPPPATTT constitutes the first potent splenocyte stimulation Dhc-lipopeptide described so far that contains only two fatty acid residues. PMID- 9222999 TI - Identification of avidin and streptavidin binding motifs among peptides selected from a synthetic peptide library consisting solely of D-amino acids. AB - Peptides consisting solely of D-amino acids (D-peptides) as opposed to their L counterparts (L-peptides) are resistant towards proteolytic degradation in the organism and may therefore be useful in future efforts to develop new stable peptide-based drugs. Using the random synthetic peptide library technique several L- and D-peptides, capable of binding to both avidin and streptavidin, were found. The L-peptides contained the previously described HPQ/M motifs, and among the D-peptides three binding motifs could be identified, of which the most frequently found one contained an N-terminal aliphatic hydrophobic amino acid (V, L or I) and an aromatic amino acid (Y or F) on the second position. At the third position in this motif several different amino acid residues were found, although N was the most frequent. Peptides representing two of the D-motifs were synthesized as well as peptides containing the HPQ/M motifs, and their binding properties were examined. Although the D-peptides were originally selected using avidin they also inhibited binding between immobilized biotin and soluble streptavidin as well as avidin. The IC50 of some of the peptides were approximately 10(5) times higher than the IC50 for biotin but some had a lower IC50 than iminobiotin. The D-peptides, which were originally selected from the library using avidin, could also inhibit the binding between streptavidin and biotin. Likewise, L-peptides selected from a library screened with streptavidin, could inhibit the binding of both streptavidin and avidin to immobilized biotin. Furthermore, the D-peptide, VFSVQSGS, as well as biotin could inhibit binding of streptavidin to an immobilized L-peptide (RYHPQSGS). This indicates that the biotin-like structure mimicked by these two seemingly very different peptides may react with the same binding sites in the streptavidin molecule. PMID- 9222998 TI - Determination of disulphide bridges in PG-2, an antimicrobial peptide from porcine leukocytes. AB - We determined the cysteine connectivity of protegrin PG-2, a leukocyte-derived antimicrobial peptide, by performing sequential enzyme digestions with chymotrypsin and thermolysin, and monitoring each digest by direct liquid chromatography-electrospray mass spectrometric analysis. This approach resolved the disulphide pairing pattern unambiguously with only picomolar amounts of PG-2. The inferred cysteine connectivity was confirmed by traditional amino acid composition analyses using nanomolar amounts of the protegrin. The results suggest that protegrins will assume a tachyplesin-like, disulphide-stabilized anti-parallel beta-sheet configuration in solution. PMID- 9223000 TI - Characterization of the conformational domains of bradykinin by computational methods. AB - The AMBER 4.0 force field was used to perform a characterization of the conformational profile of the nonapeptide bradykinin. A thorough conformational search was carried out using molecular dynamics as sampling technique, by computing cycles of high (900 K) and low (300 K) temperature trajectories. A total of 2400 minima were generated and subsequently clustered using the root mean-square of the backbone dihedral angles as criterium. After the use of a tolerance value of 20 degrees, the conformations were clustered in 233 unique conformations with energies up to 40 kcal/mol above the lowest minimum. The analysis of the low-energy conformations indicate that the peptide exhibits a high tendency to adopt a beta-turn at the C-terminus and a propensity to adopt a bent structure at the N-terminus. These results are in agreement with the experimental evidence reported in the literature and provide detailed information necessary to understand the conformational preferences of the peptide. PMID- 9223001 TI - Conformational rigidity versus flexibility in a novel peptidic neurokinin A receptor antagonist. AB - Neurokinin A receptor antagonists have been proposed as a new class of drugs for several applications in humans (asthma, intestinal motility, etc.). The rational design, synthesis, structural characterization and biological activity evaluation of a new potent, highly selective, long-lasting, peptide-based receptor antagonist are reported. The structure-activity relationship indicates that the conformational rigidity determines potency, specificity and especially the long life of the molecule in the living body. MEN10627 is the prototype of a new class of cyclic, peptide-based, neurokinin A receptor antagonists and it is a suitable candidate for clinical testing in humans. PMID- 9223002 TI - Solution versus solid-phase cyclization strategies for large sidechain lactam bridged peptides: a comparative study. AB - A 22-residue peptide with a sidechain lactam bridge involving 18 residues (60 atom cycle) has been synthesized. Three different protection schemes using Fmoc/tBu/cyclohexyl, Fmoc/tBu/allyl or Boc/Bzl/ fluorenylmethyl protecting group combinations have been explored for the solid phase of the linear precursors, which have been subsequently cyclized in solution or in the solid phase. Cyclization yields in solution have been consistently better than on solid phase; however, the solid-phase strategy requires fewer purification steps and therefore global yields are comparable. PMID- 9223003 TI - Conformational studies on synthetic peptides reproducing the dibasic processing site of pro-ocytocin-neurophysin. AB - Synthetic peptides reproducing the proteolytic processing site of pro-ocytocin were studied by different spectroscopic techniques, including circular dichroism, Fourier transform infrared absorption, and mono and bidimensional nuclear magnetic resonance, in order to ascertain the possible role of three-dimensional structure in the recognition process by maturation enzymes. Experimental results were compared with energy minimization calculations and suggest that: (i) the region situated on the N-terminus of the Lys-Arg doublet may form a beta-turn; (ii) the sequential organization of the residues participating in the beta-turn determines the privileged relative orientation of the basic amino acid sidechains and the subtype of turn; and (iii) the peptide segment situated on the C-terminal side of the dibasic doublet may assume a helix arrangement. These findings, in spite of the limitations connected to the flexibility of linear peptides, seem to substantiate the hypothesis that structural motifs around the cleavage site could be important for recognition and processing. however, a straightforward correlation between details of the secondary structure and the in vitro reactivity toward a putative convertase is not yet possible. PMID- 9223004 TI - Augmentation of the affinity of HLA class I-binding peptides lacking primary anchor residues by manipulation of the secondary anchor residues. AB - A direct binding assay has been used to investigate the effect of the secondary anchor residues on peptide binding to class I proteins of the major histocompatibility complex. Based on predictions from a previous chemometric approach, synthetic peptide analogues containing unnatural amino acids were synthesized and tested for B*2705 binding. Hydrophobic unnatural amino acids such as alpha-naphthyl- and cyclohexyl-alanine were found to be excellent substituents in the P3 secondary anchor position giving peptides with very high B*2705-binding affinity. The binding to B*2705 of peptides optimized for their secondary anchor residues, but lacking one of the P2 or P9 primary anchor residues was also investigated. Most such peptides did not bind, but one peptide, lacking the P2 Arg residue generally considered essential for binding to all B27 subtypes, was found to bind quite strongly. These findings demonstrate that peptide binding to class I proteins is due to a combination of all the anchor residues, which may be occupied also by unnatural amino acids-a necessary step towards the development of peptidic or non-peptidic antagonists for immunomodulation. PMID- 9223005 TI - Lactam bridge stabilization of alpha-helical peptides: ring size, orientation and positional effects. AB - A series of 14 residue amphipathic alpha-helical peptides, in which the sidechains of glutamic acid and lysine have been covalently joined, was synthesized in order to determine the effect of spacing, position and orientation of these lactam bridges. It was found that although an (i, i+3) spacing would position the lactam bridge on the same face of the helix, these lactams with 18 member rings were actually helix-destabilizing regardless of position or location. On the other hand, (i, i+4) lactams with 21-member rings were helix stabilizing but this was dependent on orientation. Glutamic acid-lysine lactams increased the helical content of the peptide when compared with their linear homologue in benign conditions (50 mM KH2PO4, 100 mM KCl, pH 7). Two Glu-Lys (i, i+4) lactams located at the N- and C-termini gave rise to a peptide with greater than 99% helical content in benign conditions. Peptides with Lys-Glu oriented lactams were random structures in benign conditions but in the presence of 50% TFE could be induced into a helical conformation. The stability of the single stranded alpha-helices, as measured by thermal denaturations in 25% TFE indicated that Glu-Lys oriented lactam bridges stabilized the helical conformation relative to the linear unbridged peptide. One Glu-Lys lactam in the middle of the peptide was more effective at stabilizing helical structure than two Glu-Lys lactams positioned one at each end of the molecule. The lactams with the Lys-Glu orientation were destabilizing relative to the unbridged peptide. This study demonstrates that correct orientation and position of a lactam bridge is critical in order to design peptides with high helical content in aqueous media. PMID- 9223006 TI - The application of papain, ficin and clostripain in kinetically controlled peptide synthesis in frozen aqueous solutions. AB - The capability of the cysteine proteases ficin, papain and clostripain to form peptide bonds in frozen aqueous solutions was investigated. Freezing the reaction mixture resulted in increased peptide yields in kinetically controlled coupling of Bz-Arg-OEt with various amino acid amides and dipeptides. Under these conditions, peptide yields increased up to 70% depending on the enzyme and the amino component used. Enzyme-catalysed peptide syntheses were carried out under optimized reaction conditions (temperature, amino component concentration and pH before freezing) using the condensation of Bz-Arg-OEt and H-Leu-NH2 as a model reaction. PMID- 9223007 TI - Affinity purification of 101 residue rat cpn10 using a reversible biotinylated probe. AB - The purification of large synthetic peptides using conventional separation techniques often results in poor yields and homogeneity due to the accumulation of chromatographically similar deletion and truncated impurities. We have developed a highly effective synthetic strategy and one-step purification procedure that is based on (i) the application of single coupling using HBTU/HOBt activation to reduce incomplete couplings, (ii) the use of N-(2 chlorobenzyloxycarbonyloxy)succinimide as a capping agent to terminate deletion sequences and (iii) the N-terminal derivatization of the complete peptidyl-resin with a reversible Fmoc-based chromatographic probe possessing enhanced physico chemical properties (i.e. hydrophobicity, charge or affinity label). We report the application of a biotinylated probe, activated as the succinimidyl carbonate, for the purification of a 101 residue chaperonin protein from Rattus norvegicus (rat cpn10), previously synthesized using an optimized synthetic protocol. Biotinylated rat cpn10 was separated from underivatized impurities on an immobilized monomeric avidin column. Free rat cpn10 was released from avidin agarose column with 5% aqueous triethylamine and after desalting by RP-HPLC gave 9.9% recovery. Characterization and assessment of homogeneity was achieved using ESI-MS, CZE and RP-HPLC. PMID- 9223008 TI - The immunosuppressive mini-domain of human lactoferrin. AB - It has been found that the disulphide-bridged 231-245 pentadecapeptide loop of the lactoferrin (LF) N-lobe contains a region of immunosuppressive activity. The activity resides within a thymopentin-like sequence (Arg-Lys-Pro-Val-Asp) of the loop. Peptides related to the 575-589 loop of the LF C-lobe differ in their immunomodulatory activity from those related to the 231-245 loop. We ascribe this difference to the replacement of the Asp residue in the 231-245 loop by Thr in the 575-589 loop. Two other fragments of LF which were studied, 27-34 and 309 315, do not manifest any activity in the DTH test (cellular immune response), but, on testing in vivo, stimulate the humoral immune response. The 27-34 fragment is related to the bactericidal and immunostimulative region of LF identified by Bellamy et al. [1]. Our results show that the LF molecule contains, not only the known immunostimulating mini-domain, but also a region endowed with immunosuppressive activity. PMID- 9223009 TI - Synthesis and bioactivity studies of 1-adamantanamine derivatives of peptides. AB - Small enkephalin-related peptides containing a 1-adamantanamine moiety coupled through an amide linkage at the C-terminus were synthesized. Several of the compounds showed high mu opioid activity and mu receptor selectivity. The new adamantanamine derivatives were also examined for antiviral activity against HIV 1 in a cell culture system. Some of them inhibited syncytia formation even when the antigen assay gave evidence for viral replication. PMID- 9223011 TI - Relationship of sidechain hydrophobicity and alpha-helical propensity on the stability of the single-stranded amphipathic alpha-helix. AB - The aim of the present investigation is to determine the effect of alpha-helical propensity and sidechain hydrophobicity on the stability of amphipathic alpha helices. Accordingly, a series of 18-residue amphipathic alpha-helical peptides has been synthesized as a model system where all 20 amino acid residues were substituted on the hydrophobic face of the amphipathic alpha-helix. In these experiments, all three parameters (sidechain hydrophobicity, alpha-helical propensity and helix stability) were measured on the same set of peptide analogues. For these peptide analogues that differ by only one amino acid residue, there was a 0.96 kcal/mole difference in alpha-helical propensity between the most (Ala) and the least (Gly) alpha-helical analogue, a 12.1-minute difference between the most (Phe) and the least (Asp) retentive analogue on the reversed-phase column, and a 32.3 degrees C difference in melting temperatures between the most (Leu) and the least (Asp) stable analogue. The results show that the hydrophobicity and alpha-helical propensity of an amino acid sidechain are not correlated with each other, but each contributes to the stability of the amphipathic alpha-helix. More importantly, the combined effects of alpha-helical propensity and sidechain hydrophobicity at a ratio of about 2:1 had optimal correlation with alpha-helix stability. These results suggest that both alpha helical propensity and sidechain hydrophobicity should be taken into consideration in the design of alpha-helical proteins with the desired stability. PMID- 9223010 TI - Chemical synthesis of a 7 kDa insect gonadotropic neurohormone. AB - An original insect neurohormone of 65 residues was synthesized by the solid-phase methodology using t-Boc strategy and Boc-Val-PAM-resin. The purification, conducted by several steps of liquid chromatography having mass, polarity or charge as separative criteria, yielded the product with the correct molecular weight of 6922 Da determined by mass spectrometry. The synthetic peptide had both the same affinity for the anti-native neurohormone serum and the same biological activity as the native neurohormone. PMID- 9223013 TI - A rational approach for the development of reduced-size analogues of neuropeptide Y with high affinity to the Y1 receptor. AB - Four sets of centrally truncated analogues of neuropeptide Y have been synthesized. In each series the N-terminal part was constant, while the C terminal segment was systematically varied in length. The C- and N-terminal parts were linked by 6-aminohexanoic acid. The affinity to the Y1 receptor was investigated on human neuroblastoma cells SK-N-MC. Significant differences were found between the series of peptides as well as within each set. Remarkably, the affinity did not solely depend on the length of the segment, and with increasing numbers of residues the IC50 values were not always decreased. With a given N terminal segment, only one optimal length of the C-terminal segment was found, which suggests that it is not the amino acids themselves but their 3D arrangement and orientation that is important for high receptor affinity. PMID- 9223012 TI - Conformational behaviour of a cyclolinopeptide A analogue: two-dimensional NMR study of cyclo(Pro1-Pro-Phe-Phe-Ac6c-Ile-ala-Val8). AB - The cyclic octapeptide cyclo[-Pro1-Pro-Phe-Phe-Ac6c-Ile-ala-Val8-] [C8-Ac6c], containing the Pro1-Pro-Phe-Phe sequence, followed by a bulky helicogenic C alpha,alpha-dialkylated glycine residue Ac6c [1-aminocyclohexane-1-carboxylic acid), and a D-Ala residue at position 7 has been synthesized. This cyclic peptide is a deletion analogue of the naturally occurring cyclic nonapeptide cyclolinopeptide A (CLA). It has been designed with the aim of studying the role that the Ac6c and D-Ala residues play on the conformational behaviour of the whole molecule and their influence on the conformation of the Pro1-Pro-Phe-Phe sequence when compared with cyclolinopeptide A. C8-Ac6c has been investigated in chloroform and acetonitrile solutions by 2D NMR techniques. Only one set of sharp signals is observed in both solvents. This evidence strongly supports the hypothesis that only one conformational state exists in the chosen solvents. The interpretation of the experimental data points to the existence for C8-Ac6c of a very rigid structure stabilized by intramolecular hydrogen bonds. The measured NOE effects allow the calculation of internuclear distances, which have been used as restraints in molecular dynamic calculations. The proposed conformation of the molecule shows that the Pro-Pro-Phe segment retains the conformation observed in natural CLA both in solution and in the solid state and that the Ac6c residue indeed reinforces the ring rigidity not permitting the formation of any appropriate cavity in which inorganic cations could be complexed. PMID- 9223014 TI - Sweet and bitter taste: structure and conformations of two aspartame dipeptide analogues. AB - The synthesis and X-ray diffraction analysis of two dipeptide taste ligands have been carried out as part of our study of the molecular basis of taste. The compounds L-aspartyl-D-alpha-methylphenylalanine methyl ester [L-Asp-D-(alpha Me)Phe-OMe] and L-aspartyl-D-alanyl-2,2,5, 5-tetramethylcyclopentanyl ester [L Asp-D-Ala-OTMCP] elicit bitter and sweet taste, respectively. The C-terminal residues of the two analogues adopt distinctly different conformations in the solid state. The aspartyl moiety assumes the same conformation found in other dipeptide taste ligands with the side-chain carboxylate and the amino groups forming a zwitterionic ring with a conformation defined by psi, chi 1 = 157.7 degrees, -61.5 degrees for L-Asp-D-Ala-OTMCP and 151.0 degrees, -68.8 degrees for L-Asp-D-(alpha Me)Phe-OMe. In the second residue, a left-handed helical conformation is observed for the (alpha Me)Phe residue of L-Asp-D-(alpha Me)Phe OMe with phi 2 = 49.0 degrees and psi 2 = 47.9 degrees, while the Ala residue of L-Asp-D-Ala-OTMCP adopts a semi-extended conformation characterized by dihedral angles phi 2 = 62.8 degrees and psi 2 = -139.9 degrees. The solid-state structure of the bitter L-Asp-D-(alpha Me)Phe-OMe is extended: while the crystal structure of the sweet L-Asp-D-OTMCP roughly adopts the typical L-shaped structure shown by other sweeteners. The data of L-Asp-D-(alpha Me)Phe-OMe are compared with those of its diastereoisomer L-Asp-L-(alpha Me)Phe-OMe. Conformational analysis of the two taste ligands in solution by NMR and computer simulations agrees well with our model for sweet and bitter tastes. PMID- 9223015 TI - Metal ion binding affinities of gastrin and CCK in membrane mimetic environments. AB - The fully active gastrin and CCK analogues [Nle 15]-gastrin-17 and [Nle, Thr]-CCK 9 were analysed for their Ca2+ and Tb3+ affinities in various membrane mimetic conditions. In TFE both gastrin and CCK exhibited high affinities for calcium and terbium. At saturation level identical metal ion/peptide ratios were determined with Ca2+ and Tb3+, i.e. R = 3 for gastrin and R = 1 for CCK, confirming the very similar coordination properties of the two metal ions. The conformational effects of both metal ions were found to be very similar with a disordering effect in the case of gastrin and a conformational transition to beta-turn type structure in the case of CCK. In order to mimic more properly physiological conditions, similar experiments were performed in the presence of phospholipid bilayers. No interaction of the peptides with the bilayers was observed even in the presence of mmolar Ca2+ concentrations. Induced lipid interaction via N-terminal lipo derivatization of gastrin and CCK allowed to translocate quantitatively the two hormones into phospholipid bilayers and to examine the effect of extravesicular Ca2+ on the conformation of the peptide headgroups and on their display at the water/lipid interphase. The CCK moiety of the lipo-CCK inserted into phospholipid bilayers interacts with the lipid phase and addition of Ca2+ enhances the clustering of the peptide headgroups in a more beta-sheet type conformation. Conversely, insertion of lipo-gastrin into the bilayers leads to full exposure of the gastrin headgroup to the bulk water in predominantly random coil structure. Again Ca2+ provokes aggregation. As the lipo-peptide/phospholipid system still represents only an artificial model, it remains hazardous to derive a biological relevance from these data. The significantly higher affinity of lanthanide ions than Ca2+ for the peptides could well play a role in the inhibitory activity of lanthanum on the signal transduction of the CCK family of hormones. PMID- 9223016 TI - Use of a gel-forming dipeptide derivative as a carrier for antigen presentation. AB - A dipeptide of the formula Fmoc-Leu-Asp and some other related dipeptides were synthesized in solution by standard methods. When such peptides are dissolved in water at concentrations below 1% at 100 degrees C and cooled below 60 degrees C they form turbid solutions and eventually viscoelastic gels at lower temperatures. Such gels are thermoreversible and can also be disrupted by mechanical agitation. At a concentration of 2 mg/ml the peptide Fmoc-Leu-Asp forms an aqueous gel at 60 degrees C with a shear modulus of 80 Pa measured at a frequency of 1 rad/s. Peptide solutions undergo an abrupt increase in light scattering between 1 and 1.5 mg/ml at both 23 and 60 degrees C. By analogy with previous observations of other systems, this increase appears to be due to the formation of filamentous micelles and the aggregation of filaments into a three dimensional network. When low molecular weight adamantanamine derivatives, which are inherently non-antigenic antiviral drugs, were incorporated into the Fmoc-Leu Asp gel and injected into rabbits, high titre specific antibodies were efficiently produced without the need for additional adjuvant. Both the physical properties of the gel and its effect on the antigenicity of low molecular weight compounds suggest a number of practical applications. PMID- 9223017 TI - Synthesis and biological characterization of a series of analogues of omega conotoxin GVIA. AB - The 27-residue polypeptide omega-conotoxin GVIA (omega-CgTx), from the venom of the cone shell Conus geographus, blocks N-type neuronal calcium channels. It contains three disulphide bridges. We report here the synthesis and biological characterization of a series of analogues in which one disulphide has been replaced by substitution of appropriate Cys residues with Ser, viz. [Ser1,16] omega -CgTx, [Ser8,19]-omega-CgTx, [Ser15,26)-omega-CgTx, [Ser16]-omega-CgTx8-27 and [Ser15]-omega-CgTx1-19. All syntheses were conducted manually using either Boc or Fmoc methodology. Deprotected peptides were oxidized to their bridged forms using either aerial oxidation or aqueous dimethyl sulphoxide. Peptides were purified using RP-HPLC, and their purity and identity were checked by RP-HPLC, capillary electrophoresis and mass spectrometry. Inhibition of neuronal N-type calcium channels was assessed as the inhibition of the twitch responses of rat vas deferens stimulated with single electrical pulses at 20 second intervals. None of these analogues was biologically active, suggesting that the disulphides play an important role in maintaining biological activity. PMID- 9223019 TI - Inversion of 3(10)-helix screw sense in a (D-alpha Me)Leu homo-tetrapeptide induced by a guest D-(alpha Me)Val residue. AB - The terminally blocked tetrapeptide pBrBz-[D-(alpha Me)Leu]2-D-(alpha Me)Val-D (alpha Me)Leu-OfBu is folded in the crystal state in a left-handed 3(10)-helical structure stabilized by two consecutive 1<--4 C = O...H-N intramolecular H-bonds, as determined by X-ray diffraction analysis. A CD study strongly supports the view that this conformation is also that largely prevailing in MeOH solution. A comparison with the published conformation of pBrBz-[D-(alpha Me)Leu]4-OfBu indicates that incorporation of a single internal beta-branched (alpha Me)Val guest residue into the host homo-tetrapeptide from the gamma-branched (alpha Me)Leu residue is responsible for a dramatic structural perturbation, i.e. an inversion of the 3(10) screw sense from right to left-handed. PMID- 9223018 TI - Application of the multiple antigenic peptides (MAP) strategy to the production of prohormone convertases antibodies: synthesis, characterization and use of 8 branched immunogenic peptides. AB - Antiserum against an N-terminal sequence of murine prohormone convertase-1 (mPC1) incorporating the sequence immediately following the junction between the putative pro-region and the active enzyme was obtained. This was accomplished using the multiple antigenic peptide (MAP) approach whereupon an 8-branched polylysine core to which are grafted multiple copies of a 16 amino acid peptide representing the N-terminal sequence of mPC1 (positions 84-99) was synthesized by solid-phase Fmoc chemistry. The ensuing peptide was purified and fully characterized by RP-HPLC, 1H-NMR, amino acid composition, peptide sequencing and ion-spray mass spectrometry. The immunological properties of the resulting antibodies in detecting recombinant PC1 in both crude and purified preparations were compared with antibodies raised against a similar N-terminal segment of PC1 but using the conventional method of peptide-carrier protein conjugation and also developed against a C-terminal fusion protein of PC1. Our data indicate that the MAP antibody was as efficient as both the amino and carboxy-terminal antibodies in qualitative as well as quantitative analysis of PC1 encoded protein by radioimmunoassay. Following an identical approach, antibodies against other prohormone convertases like furin, PC5/6 and PACE4 were also developed and subsequently applied to a number of biochemical and immunological studies. In each case, the case of preparation and high immunogenicity of the MAP approach were confirmed and reside in the simplicity and rapidity with which a potent and useful antiserum is obtained. PMID- 9223021 TI - Effect of Na+ current on excitation-contraction coupling in ventricular myocytes of guinea pig heart. AB - We investigated the effect of Na+ current on the Ca2+ current and Ca2+ transients in cardiac myocytes. Myocytes were isolated from the ventricles of guinea-pig hearts by enzymatic dispersion. The membrane currents were recorded by the whole cell voltage clamping. The Ca2+ current was activated by depolarisation from -80 to +5 mV preceded by the prepulses to -40 mV. Cellular action potentials (APs) were recorded by current clamping. Intracellular [Ca2+] was assessed by recording of fluorescence of Indo-1 loaded into cells. In current clamped cells (APs recorded) 20 microM tetrodotoxin (TTX) reduced the time to 75% of amplitude of Ca2+ transients from 50 +/- 6.6 ms to 32 +/- 5 ms (n = 7). In voltage clamped cells prepulses from the holding potential of -80 mV to -40 mV 50-100 ms long activated the Na+ current and initiated step increase in [Ca2+] reaching 30-50% of the total amplitude of the transient. Prepulses 10-20 ms long initiated increase in [Ca2+] merging with that elicited by Ca2+ current into smooth rising phase. Blocking of Na+ current with TTX or by switching the holding potential from -80 to -40 mV increased the amplitude of the Ca2+ current by 38 +/- 3.2% (n = 8) and 43 +/- 9% (n = 7), respectively, and eliminated the initial step increase in [Ca2+]. When 10-20 ms prepulses were used, blocking of Na+ current with TTX or switching of the holding potential decreased the time to 75% of amplitude of Ca2+ transients from 27 +/- 3.7 ms to 12 +/- 1.2 ms (n = 5) and from 25 +/- 3.1 ms to 14 +/- 1.1 ms (n = 9), respectively. 100 microM Cd2+ inhibited the initial rise in [Ca2+], however, the inhibition did not correlate with degree of inhibition of Ca2+ current. The Na+ current activated prior to Ca2+ current reduces its amplitude and decreases the rate of release of Ca2+ from sarcoplasmic reticulum. In voltage clamped cells this could result from Ca2+ influx prior to onset of Ca2+ current initiated by Na+ current escaping from the voltage control and/or reversal of Na/Ca exchange due to increase in subsarcolemmal [Na+]. In cells in which Ca2+ transients were initiated by APs only the second mechanism is conceivable. PMID- 9223020 TI - The immune-neuro-endocrine interactions. AB - This article reviews data concerning the interactions between immune, endocrine and neural systems in physiological, pathophysiological and stress conditions in animals and humans. Numerous studies have provided evidence that these systems interact with each other in maintaining homeostasis. This interaction may be classified as follows: immune, endocrine and neural cell products coexist in lymphoid, endocrine and neural tissue. Endocrine and neural mediators modulate immune system activity. Immune, endocrine and neural cells express receptors for cytokines, hormones, neuropeptides and transmitters. PMID- 9223022 TI - The adrenal-renal vascular connection contributes to increase in renal vascular resistance during an experimental hypotension in the rat. AB - The adrenal vascular connection (ARVC) was described for the first time in the cat by Cow (1914) and by other authors in the dog, rat, rabbit and humans. The aim of the present study is to investigate the role of above connection in regulation of renal vascular resistance (RVR), and renal blood flow (RBF) during decrease in blood pressure in the rat. Animals were divided into three groups. In the first group, mean arterial pressure (MAP) was unchanged. In the second and the third group MAP was maintained at 50 mmHg. In addition in the third group, an alpha adrenergic receptor blockade was produced with intravenous infusion of phentolamine. After stabilisation of RBF, in all groups, the tissue between the adrenal gland and the kidney was cut. RBF and MAP were measured and recorded. In the first and the third group, the elimination of ARVC neither influenced RBF nor RVR. In the second group the elimination of ARVC caused increase in RBF and decrease in RVR (p < 0.01). Results of the present study provide the evidence that catecholamines reaching the kidney, directly from the adrenal gland through ARVC, during the severe hypotension are responsible for reducing of renal blood flow and increase in renal vascular resistance in the rat. PMID- 9223023 TI - Study with two prokinetics in functional dyspepsia and GORD: domperidone vs. cisapride. AB - The HT4-agonist Cisapride (CIS) and the peripheral D2-antagonist Domperidone (DOMP) have distinct prokinetic actions. We compared their clinical efficacy in 127 dyspeptic patients. Patients with upper abdominal complaints of > 1 month duration, who had a normal UGE were allocated to the REFLUX-group (RG), (predominance of heartburn, acid regurgitation or retrosternal pain) or if devoid of this specific symptomatology to the DYSPEPSIA-group (DG) In a double-blind randomised fashion and allocated to 10 mg CIS or 20 mg DOMP qid (RG) or tid (DG) for 1 month and followed-up for further 2 months. In RG (N = 43, p < 0.05) the response rates were clearly in favour of CIS, but not in DG (N = 84). In RG DOMP was more effective against nausea. The benefit of both therapies was largely maintained in the follow-up period. Cisapride and domperidone were effective in the treatment of dyspepsia. Cisapride was more effective than domperidone in the REFLUX-Group. PMID- 9223024 TI - Effect of acceleration stress on salivary cortisol and plasma cortisol and testosterone levels in cadet pilots. AB - The effects of acceleration (Gz) on changes in the levels of cortisol in saliva and of cortisol and testosterone in serum have been studied in 48 cadet pilots exposed to a linear acceleration gradient (0.2 G/s) until a loss of coordination when the mean G value was found to be 5.94 +/- 0.57. Three patterns of salivary cortisol responses were discerned based on Gz-induced significant changes: increase (I; n = 20), decrease (D; n = 8), the magnitude of changes being dependent on the pre-Gz values. Fifteen min after the Gz load, the mean salivary cortisol was significantly higher from the pre-Gz value in all subjects combined. In 19 subjects, who consented to blood sampling, significant increases in serum cortisol were observed both 3 and 15 min post-Gz (by 37 and 57% respectively) while, a significant increase in serum testosterone concentration has been observed only 3 min post-Gz. Testosterone levels 3 min post-Gz were significantly correlated with the final Gz values (r = 0.54; p < 0.05). A significant correlation was also observed between all salivary and serum cortisol values combined (r = 0.696; p < 0.001). It has been concluded that acceleration stress, although of very short duration, proved very potent in eliciting glucocorticoid and androgen responses. PMID- 9223025 TI - Endogenous nitric oxide in the control of esophageal motility in humans. AB - Recent animal studies have suggested that nitric oxide (NO) plays an important role in the regulation of esophageal motility, being partly responsible for latency period and latency gradient between the onset of a swallow and contractions of esophageal circular smooth muscles. The aim of this study was to evaluate whether endogenous NO is responsible for physiological timing of forthcoming contractions in the human esophageal body after swallowing. Eight male volunteers (age 21-25 years, weight 67-82 kg) were involved in this placebo controlled study on the effects of increasing doses of the NO synthase blocker, NG-monomethyl-L-arginine (L-NMMA 1.0-4.0 mumol/min i.v.), and/or L-arginine (L arg) (30 mumol/kg-min i.v.) on the peristalsis of esophageal body in response to wet swallows (5 ml of water) and lower esophageal sphincter (LES) resting pressure. The esophageal motor activity was determined manometrically using 3 channel electronic catheter. Additionally, during all examinations arterial blood pressure (BP) was measured every 5 min. L-NMMA resulted in a significant and dose dependent reduction in the latency period between swallows and the onset of contractions which was most pronounced in the distal esophagus (control: 7.07 +/- 0.74 s vs. L-NMMA 4.0 mumol/min: 5.87 +/- 0.57 s), and this effect was partially reversed after addition of L-arg to the L-NMMA infusion (6.91 +/- 0.62 s). L-NMMA infusion significantly reduced the duration of contractions and increased the velocity of onset propagation but did not change the amplitude of contractions and again, these effects were reversed during simultaneous infusion of L-arg. The resting tone of LES increased significantly during infusion of L-NMMA and these effects were reversed by addition of L-arg. The mean BP significantly increased during infusion of L-NMMA (control 97.0 +/- 5.7 vs. L-NMMA 4.0 mumol/min: 116.4 +/- 3.1 mm Hg) and this was also reversed by L-arg. We conclude that in humans endogenous NO is involved, at least in part, in the physiological regulation of motility patterns of the distal esophageal body and LES. PMID- 9223027 TI - Increase in cardiodepressant factor release from the posterior pituitary lobe after angiotensin II infusion into the internal carotid artery. AB - In our previous research the presence of a cardiodepressant factor in the medium incubating the posterior pituitary lobe 'in situ' has been demonstrated. This study presents experiments demonstrating cardiodepressant activity in medium incubating the posterior pituitary lobe 'in situ' and in dialysates of venous blood from the sella turcica region before and during angiotensin II (ANG II) infusion into the internal carotid artery in rat. Cardiodepressant activity was determined on spontaneously discharging isolated auricle of the right atrium in a two-day-old rat. It has been demonstrated that medium incubating the posterior pituitary lobe which was collected during angiotensin II infusion caused a greater decrease in auricle discharge rate than medium collected before the infusion. Angiotensin II infusion into the internal carotid artery caused a dose dependent increase in cardiodepressant activity in dialysates of blood outflowing from the sella turcica region. The present results indicate that angiotensin II increase the release of cardiodepressant factor from the posterior pituitary lobe into blood in a dose-dependent manner. PMID- 9223026 TI - Metabolic alkalosis induced by pre-exercise ingestion of NaHCO3 does not modulate the slow component of VO2 kinetics in humans. AB - Seven healthy physically active nonsmoking men, aged 22.4 +/- (SD) 1.8 years performed two 6 min bouts of cycling at 40% VO2max (sub-lactate threshold/low power output exercise) and 87% VO2max (supra-lactate threshold/high power output exercise) at 70 rev.min-1, separated by 20 minutes rest, on two occasions: once as a control experiment (test C) and on a different day at approximately 1.5 h after ingestion of 250 mg (3 mmol).(kg body weight)-1 of NaHCO3 (test A). At the onset of low and high power output exercise performed after ingestion of NaHCO3, antecubital venous blood pH and HCO3- were significantly elevated (p < 0.05). Moreover, blood pH and HCO3-, tested at every minute of low and high power output exercise, was significantly higher (p < 0.05) in test A than in test C. No difference was found in plasma lactate concentration [La]pl during low power output exercise between A and C tests. In the terminal phase of the high power output exercise (87% VO2max) the level of [La]pl rose more rapidly in test A than in test C, reaching in the sixth minute of cycling 8.27 +/- 1.11 and 6.76 +/- 0.68 mmol.l-1 (p < 0.01) in test A and C, respectively. No significant differences were found in the rate of VO2 measured breath-by-breath between A and C tests, both during low and high power output exercise. The slow component of VO2 kinetics (expressed by difference between VO2 measured at the 6th minute of exercise minus the VO2 reached at the 3rd minute), occurring only during exercise corresponding to 87% VO2, was not significantly different in C and A tests (0.373 +/- 0.050 and 0.339 +/- 0.078 1 O2, respectively). The total VO2 consumed throughout the six minute cycling at power output of 40 and 87% VO2max performed in control conditions and after ingestion of NaHCO3 was not significantly different. We have demonstrated that significantly reduced exercise acidemia accompanied by a significantly elevated level of [La]pl accumulation, did not affect the slow component of the VO2 kinetics and the magnitude of oxygen uptake during exercise corresponding to 40 and 87% of VO2max. PMID- 9223028 TI - The effect of drugs acting on CCK receptors and rat free exploration in the exploration box. AB - The effects of cholecystokinin (CCK) CCKA receptor antagonist devazepide (10 micrograms/kg and 1.0 mg/kg), CCKB receptor antagonist L 365260 (1.0 mg/kg), and CCKB receptor agonist CCK tetrapeptide (CCK-4, 75 micrograms/kg), and their concomitant administration with antidepressants desipramine (10 mg/kg) and citalopram (10 mg/kg) on rat exploratory behaviour were studied in the recently developed exploration box test. In addition, the effects of repeated administration of despiramine (10 mg/kg) and citalopram (10 mg/kg) were studied. After acute administration, CCK-4 decreased significantly the number of line crossings, rears, investigative approaches, and the time spent exploring. The time of latency and the number of entries into large arena were unchanged. Desipramine reduced all observed criterions of rat behaviour, but citalopram was ineffective. Devazepide (1.0 mg/kg) and L 365260 (1.0 mg/kg) had no effect on rat behaviour after acute or repeated administration. L 365260 (1.0 mg/kg) blocked the antiexploratory effect of CCK-4, whereas devazepide (10 micrograms/kg) did not. No interaction of CCK-4, devazepide, or L 365260 treatment with antidepressant treatment was found. Our results suggest that the administration of a CCKB agonist diminishes rat exploratory behaviour, but neither CCKA nor CCKB receptor blockade induces changes on rat exploratory behaviour in the free exploration paradigm. PMID- 9223029 TI - The effects of flumazenil, Ro 15-4513 and beta-CCM on the behaviour of control and stressed mice in the plus-maze test. AB - The effects of the benzodiazepine receptor antagonist flumazenil (Ro 15-1799), the benzodiazepine receptor partial inverse agonist Ro 15-4513 and the benzodiazepine receptor inverse agonist beta-CCM on the behaviour of control and small platform stressed mice studied. Small platform stress was induced by placing the animals on small platforms (d = 3.5 cm) surrounded by water for 24 hours. This technique involves several factors of stress such as rapid eye movement sleep-deprivation, isolation, immobilization, falling into the water and soaking. In the plus-maze test small platform stress induced changes indicating anxiolytic action-an increase of the percentage of entries made onto and the percentage of time spent on the open arms. In control mice flumazenil (2.0 and 10.0 mg/kg), Ro 15-4513 (0.5; 1.0; 2.5; 5.0 and 10.0 mg/kg), and beta-CCM (1.0 and 2.0 mg/kg) exerted dose-dependent anxiogenic effect. The small platform stress induced an enhancement of the anxiogenic effect of flumazenil, but not that of Ro 15-4513 and beta-CCM. The selective enhancement of flumazenil's action may be explained with the mode of action of flumazenil. It is proposed that small platform stress causes changes in the concentration of the endogenous benzodiazepine receptor ligand with stress protective activity and flumazenil acts by blocking the effects of this endogenous ligand. PMID- 9223030 TI - ACTH 4-9 analogue facilitates the antiimmobility effect of antidepressants and dopamine agonists in swimming rats. AB - Evidence exists that the 4-10 or 4-9 fragments of adrenocorticotropic hormone (ACTH) produce some behavioral effects in animals and in humans. The present study was designed to investigate whether ACTH 4-9 interferes with the effects of antidepressants: fluoxetine (FLU), fluvoxamine (FOX), selegiline (SEL) or dopamine agonists: piribedil (PRB) or quinpirol (QPR) in forced swimming test and in open field in rats. ACTH 4-9 was given in a single dose (25, 50 or 100 micrograms/kg) or for 7 days (50 micrograms/kg/day), alone or together with antidepressants or dopamine agonists. It was shown that ACTH 4-9 alone did not influence the behavior of rats. However, when given in a single dose, ACTH 4-9 potentiated the antiimmobility effect of all antidepressants and dopamine agonists. ACTH 4-9 given for 7 days, facilitated only the effect of selegiline. The results suggest a functional interaction of ACTH 4-9 with serotonergic and dopaminergic brain mechanisms of drugs action. PMID- 9223031 TI - Mediation by nitric oxide of the carbachol-induced corticosterone secretion in rats. AB - Nitric oxide synthase, an enzyme responsible for nitric oxide (NO) formation has been found in the hypothalamic paraventricular nucleus and median eminence, structures closely associated with regulation of the pituitary activity, and the pituitary gland itself. Nitric oxide modulates the stimulated release of CRH from the rat hypothalamus in vitro, which suggests its role in regulating the secretion of ACTH from the pituitary corticotrops and of corticosterone from the adrenal cortex. The purpose of the present study was to elucidate the yet unknown role of endogenous NO in the HPA response to central cholinergic stimulation in conscious rats. Neither L-arginine an NO precursor, nor the NO synthase blockers N omega-nitro-L-arginine methyl ester (L-NAME) and N omega-nitro-L-arginine (L NNA) caused any consistent changes in the basal serum corticosterone levels. L arginine, given in higher doses (120-150 mg/kg ip) 15 min prior to icv carbachol (2 micrograms), markedly diminished the carbachol-induced rise in corticosterone secretion. Systemic pretreatment with the nitric oxide synthase inhibitor L-NAME (5 mg/kg) significantly raised the carbachol-elicited corticosterone response, while addition of L-arginine completely blocked the effect of L-NAME. A similar increase in the carbachol-induced corticosterone response was produced by icv pretreatment with L-NAME (2 micrograms), indicating a central site of the NO interaction with cholinergic stimulation of the HPA response. L-NAME is a weak inhibitor of neuronal NOS itself, and must first be de-estrified to N omega-nitro L-arginine to potently inhibit this enzyme. Systemic (10 mg/kg) and icv (1 microgram) pretreatment with L-NNA enhanced more effectively the carbachol induced rise in corticosterone secretion than did pretreatment with L-NAME by either route. These results are the first direct evidence that endogenous NO significantly inhibits the HPA response to central cholinergic, muscarinic receptor stimulation under in vivo conditions. PMID- 9223032 TI - Serious technology assessment for health care information technology. AB - United States health care is engaged in an ambitious project to make its clinical and administrative records "100% electronic." Substantial benefits are expected in both clinical care delivery and medical research (especially for public health surveillance and outcomes/effectiveness studies). Substantial costs also potentially accrue, beyond the large outlays for an expanded computer and telecommunications infrastructure. Privacy and confidentiality are obviously at risk if such systems cannot be made secure. Limited empirical evidence currently available suggests health information systems security may not be very good, at least in the "average" institutional setting. Privacy-focused critics of electronic record-keeping are sometimes accused of taking Luddite stands, insufficiently attentive to IT's benefits. It may also be fair to worry about a certain Panglossian tendency in "industry" commentary, insufficiently attentive to potential problems. Better federal and state laws structuring health data use will help; the industry must also attend more candidly to the technical uncertainties. PMID- 9223034 TI - Beyond the superhighway: exploiting the Internet with medical informatics. AB - As in other areas of society, the Internet and the World Wide Web are becoming important topics in medical informatics. This is evident from the recent American Medical Informatics Association's 1996 Annual Fall Symposium, where the theme was "Beyond the Superhighway: Exploiting the Internet with Medical Informatics." Of the over 330 papers and abstracts published in the Proceedings, one third dealt with the Internet and/or the Web. In some cases, system developers demonstrated how this technology can do old tasks in new ways. In other cases, researchers described new tasks that are now possible with this technology. Still others examined this technology to show how it can be evaluated and improved. This paper summarizes their accomplishments. PMID- 9223033 TI - An industrial process view of information delivery to support clinical decision making: implications for systems design and process measures. AB - Clinical decision making is driven by information in the form of patient data and clinical knowledge. Currently prevalent systems used to store and retrieve this information have high failure rates, which can be traced to well-established system constraints. The authors use an industrial process model of clinical decision making to expose the role of these constraints in increasing variability in the delivery of relevant clinical knowledge and patient data to decision making clinicians. When combined with nonmodifiable human cognitive and memory constraints, this variability in information delivery is largely responsible for the high variability of decision outcomes. The model also highlights the supply characteristics of information, a view that supports the application of industrial inventory management concepts to clinical decision support. Finally, the clinical decision support literature is examined from a process-improvement perspective with a focus on decision process components related to information retrieval. Considerable knowledge gaps exist related to clinical decision support process measurement and improvement. PMID- 9223036 TI - Rapid approximation of confidence intervals for Markov process decision models: applications in decision support systems. AB - OBJECTIVE: Develop the methodological foundation for interactive use of Markov process decision models by patients and physicians at the bedside. DESIGN: Monte Carlo simulation studies of a decision model comparing two treatments for benign prostatic hypertrophy: watchful waiting (WW) and transurethral prostatectomy (TUR). MEASUREMENTS: The 95% confidence interval (CI) for the mean of the Markov model; the correlation of a linear approximation with the full Markov model; the predictive performance of the approximation; the information index of specific utilities in the model. RESULTS: The 95% CI for the gain in utility with initial TUR was -1.4 to 19.0 quality-adjusted life-months. A multivariate linear model had an excellent fit to the predictions of the Markov model (R2 = 0.966). In an independent data set, the linear model also had a high correlation with the full Markov model (R2 = 0.967); its predictions were unbiased (p = 0.597, paired t test); and, in 96.4% of simulated cases, its treatment recommendation was the same. CONCLUSION: Using the linear model, it was possible to efficiently compute which health state had the largest contribution to the variance of the decision model. This is the most informative utility value to elicit next. The most informative utility at any point in a sequence changed depending on utilities previously entered into the model. A linear model can be used to approximate the predictions of a Markov process decision model. PMID- 9223035 TI - Consumer informatics in chronic illness. AB - OBJECTIVE: To explore the informatic requirements in the home care of chronically ill patients. DESIGN: A number of strategies were deployed to help evoke a picture of home care informatics needs: A detailed questionnaire evaluating informational needs and assessing programmable technologies was distributed to a clinic population of parents of children with cancer. Open ended questionnaires were distributed to medical staff and parents soliciting a list of questions asked of medical staff. Parent procedure training was observed to evaluate the training dialog, and parents were observed interacting with a prototype information and education computer offering. RESULTS: Parents' concerns ranged from the details of managing day to day, to conceptual information about disease and treatment, to management of psychosocial problems. They sought information to solve problems and to provide emotional support, which may create conflicts of interest when the material is threatening. Whether they preferred to be informed by a doctor, nurse, or another parent depended on the nature of the information. Live interaction was preferred to video, which was preferred to text for all topics. Respondents used existing technologies in a straightforward way but were enthusiastic about the proposed use of computer technology to support home care. Multimedia solutions appear to complement user needs and preferences. CONCLUSION: Consumers appear positively disposed toward on-line solutions. On-line systems can offer breadth, depth and timeliness currently unattainable. Patients should be involved in the formation and development process in much the same way that users are involved in user-centered computer interface design. A generic framework for patient content is presented that could be applied across multiple disorders. PMID- 9223038 TI - Medical informatics challenges of the 1990s: acknowledging secular change. PMID- 9223037 TI - Using computer-based medical records to predict mortality risk for inner-city patients with reactive airways disease. PMID- 9223039 TI - An fMRI study of the human cortical motor system response to increasing functional demands. AB - Functional magnetic resonance imaging (fMRI) was used to study activation changes in the human primary motor-sensory areas (MAs), supplementary motor areas (SMAs), premotor areas (PMAs) and the superior and inferior parietal areas (SPAs, IPAs) during right hand finger movements as the rate, force and complexity of movement were varied. A preliminary reproducibility study of a single subject doing the same repetitive index finger movements in nine different sessions over a six week period demonstrated highly consistent and highly localized activation in the contralateral MA. ANOVAs demonstrated highly significant main effects of increasing the force and complexity of movement, thereby illustrating the distributed and integrated systemic character of the cortical motor system. Interactions between brain region and the rate and complexity of movements suggested functional specialization of some components of the system. Increasing the rate of movement led to increased activity only in the contralateral MA; increasing complexity led to greater increases in activity in the left and right SPAs and the left IPA than in other areas. Although activation was evident in varying degree throughout the multiple motor areas, only the MAs showed consistent lateralization of activation. PMID- 9223040 TI - Chiari malformation type I: a new MRI classification. AB - Thirty patients with Chiari I malformation were examined by MRI over 2-year period. All patients underwent MRI scan before and after surgical decompression of the posterior fossa. Images of the craniocervical junction confirmed tonsillar herniation in all cases and allowed the definition of two anatomically distinct types of Chiari malformation. Twenty-one of the 30 patients (70%) had concomitant syringomyelia and were classified as type A, while the remaining 9 patients (30%) had evidence of frank herniation of the cerebellar tonsils below the foramen magnum without evidence of syringomyelia and were labeled type B. Type A patients had a predominant central cord symptomatology; type B patients exhibited signs and symptoms of brain stem or cerebellar compression. The concomitant cord cavitary lesions (syringomyelia) were noncommunicating (isolated syrinxes), which were separated from the fourth ventricle by a syrinx-free segment of normal spinal cord. Holocord hydromyelic cavities were seen in 8 out of 21 patients with syringomyelia, isolated cervical cavities were seen in 4 patients, while combined cervical and thoracic cavities were seen in 9 patients. Kinking of the medullocervical junction and brain stem was seen in 20 out of 30 patients (67%). MRI has proved to be an excellent, noninvasive means of studying of the craniocervical anatomy; it has allowed a classification of Chiari malformation based on objective anatomic criteria with prognostic and clinical relevance. PMID- 9223041 TI - Fast flair imaging of the brain using the fast spin-echo and gradient spin-echo technique. AB - The purpose of this study was to compare the gradient spin-echo (GRASE) to the fast spin-echo (FSE) implementation of fast fluid-attenuated inversion recovery (FLAIR) sequences for brain imaging. Thirty patients with high signal intensity lesions on T2-weighted images were examined on a 1.5 T MR system. Scan time minimized thin-section FLAIR-FSE and FLAIR-GRASE sequences were obtained and compared side by side. Image assessment criteria were lesion conspicuity, contrast between different types of normal tissue, image quality, and artifacts. In addition, contrast ratios and contrast-to-noise ratios were determined. Compared to FSF, the GRASE technique allowed a 17% reduction in scan time but conspicuity of small lesions in particular was significantly lower on FLAIR-GRASE images because of higher image noise and increased artifacts. Gray-white differentiation was slightly worse on FLAIR-GRASE. Physiological ferritin deposition appeared slightly darker on FLAIR-GRASE images and susceptibility artifacts were stronger. Fatty tissue was less bright with FLAIR-GRASE. With current standard hardware equipment, the GRASE technique is not an adequate alternative to FSE for the implementation of fast FLAIR sequences in routine clinical MR brain imaging. PMID- 9223042 TI - Measurement of liver blood volume using a macromolecular MRI contrast agent at equilibrium. AB - Liver regional blood volume (LRBV) is altered by several disease states and various drugs. Preliminary studies in the rat, using research MR imaging instruments at 2T and vascular contrast agents, have suggested that MRI may be used to measure LRBV. Our goal was to develop a technique for measuring LRBV using a clinical machine at 1.5 T. This study was performed in the rabbit, using CarboxyMethylDextran Gd-DTPA, a macromolecular contrast agent with a molecular weight of 158 kDa. MRI was performed at 1.5 T, in the plane of the inferior vena cava, with and without flow compensation, before contrast injection and in the steady state after injection. Accuracy and stability of LRBV measurement, over 2 h and with various doses (0.01-0.05 mmol/kg), was tested against a standard Evan's Blue dye-indicator technique. LRBV was 28 +/- 2 mL/100 g when measured by MRI with flow compensation, which is in good agreement with the literature and with the 26 +/- 6 mL/100 g, measured by the Evan's Blue dilution technique. Measurements varied less than 7% over time and less than 9% over the range of doses. LRBV was overestimated using a sequence without flow compensation especially when large doses of contrast agent were injected. This noninvasive MRI technique provides a simple method for measuring liver LRBV and offers new prospects for future physiological and pathological studies. PMID- 9223043 TI - Intracranial meningeomas: time- and dose-dependent effects of irradiation on tumor microcirculation monitored by dynamic MR imaging. AB - The purpose of this study was the characterization of the time- and dose dependent effects of irradiation on tumor microcirculation by means of dynamic MR imaging and correlation of the estimated data with tumor response in patients with meningeomas. Dynamic MR imaging studies were performed in 20 patients with intracranial meningeomas prior to (n = 20) and at 6 (n = 17), 18 (n = 17), and 50 wk (n = 14) after the end of radiotherapy. In seven of these patients, dynamic measurements were also performed during fractionated radiotherapy after approximate 20 Gy and 54 Gy. During and after short-time infusion of gadopentetate dimeglumine, the kinetics of lesion response was resolved using a strongly T1-weighted saturation recovery TurboFLASH (SRTF) sequence. The signal time courses of the suspected lesions were analyzed using a pharmacokinetic two compartment model. The calculated parameters amplitude A (reflecting gadopentetate dimeglumine accumulation in the extracellular space) and exchange rate constant k21 (depending on vascular permeability and blood flow) were displayed as color-coded images and analyzed as a function of time of therapy and radiation dose. All meningeomas showed a high exchange rate constant k21 (median, 5.7 min-1; range, 1.9-23.0 min-1) and a high amplitude A (median, 1.5 arbitrary units; range, 1.1-2.7) prior to X-ray treatment. During radiotherapy we found a dose related significant (p < .01) increase of k21 accompanied by an increase of the amplitude A as compared to the pretreatment values. Analysis of tumor volume 6, 18, and 50 wk after X-ray treatment revealed two different groups. In the responder group (n = 13) the median of the tumor volume decreased from 10.0 to 7.5 cm3. For this group, we found a significant drop (p < .01) of the median of the amplitude A and a decrease of the exchange rate constant k21. In the nonresponder group (n = 4) the median of the tumor volume increased after radiation from 3.5 to 4.5 cm3. The pharmacokinetic analysis revealed a decrease of the amplitude A-and an increase of the exchange rate constant k21. The response of meningeomas to radiotherapy is influenced by the effect of X-rays on tumor microcirculation. This effect on tumor microcirculation can be derived by analysis of pharmacokinetic maps obtained from dynamic MR images. Furthermore, these pharmacokinetic maps can possibly be used to differentiate groups of patients who respond or do not respond to radiotherapy and, thus, could benefit from another treatment modality. PMID- 9223044 TI - Anisotropic water diffusion in white and gray matter of the neonatal piglet brain before and after transient hypoxia-ischaemia. AB - Measurements of tissue water apparent diffusion coefficient (ADC) performed with diffusion sensitization applied separately along the x, y, and z axes revealed significant diffusion anisotropy in both cerebral white and gray matter in six newborn (< 24 h old) piglets. Mean baseline white matter ADC for a particular region of interest was 125.8% (SD 32.0%; p < .001) greater when the diffusion gradients were applied along the y axis as compared to along the x. For the cortical gray matter region considered, the situation was reversed, the mean ADC value measured along x exceeding that along y by 15.2% (SD 6.1%; p < .01). Forty three hours subsequent to a transient cerebral hypoxic-ischaemic insult, phosphorous MRS measurements indicated that the animals had suffered severe secondary cerebral energy failure. This was accompanied by a significant (p < .01) decrease in the white matter anisotropy, such that the mean y direction ADC now exceeded that along the x by only 70.9% (SD 29.4%; p < .03). There was no change in the gray matter anisotropy. The average of the ADC values measured in the x, y, and z directions had decreased by 35.3% (SD 18.5%; p < .01) in white matter and 31.4% (SD 21.9%; p < .05) in cortical gray matter. Diffusion anisotropy measurements may provide additional information useful in the characterisation of hypoxic-ischaemic injury in the neonatal brain, and must be considered if tissue water ADC values are to be unambiguously interpreted in this context. PMID- 9223045 TI - Diffusion anisotropy in excised normal rat spinal cord measured by NMR microscopy. AB - A conventional spin-echo NMR imaging pulse sequence was used to obtain high resolution images of excised normal rat spinal cord at 7 and 14 T. It was observed that the large pulsed-field gradients necessary for high-resolution imaging caused a diffusion weighting that dominated the image contrast and that could be used to infer microscopic structural organization beyond that defined by the resolution of the image matrix (i.e., fiber orientation could be assigned based on diffusion anisotropy). Anisotropic diffusion coefficients were therefore measured using apparent diffusion tensor (ADT) imaging to assess more accurately fiber orientations in the spinal cord; structural anisotropy information is portrayed in the six unique images of the complete ADT. To reduce the dimensionality of the data, a trace image was generated using a separate color scale for each of the three diagonal element images of the ADT. This new image retains much of the invariance of the trace to the relative orientations of laboratory and sample axes (inherent to a greyscale trace image) but provides, by the use of color, contrast reflecting diffusion anisotropy. The colored trace image yields a pseudo-three-dimensional view of the rat spinal cord, from which it is possible to deduce fiber orientations. PMID- 9223046 TI - A multidimensional partition analysis of SSFP image pulse sequences. AB - The k-space description, of MRI pulse sequences, has been combined with a partition model in order to model the image reconstruction and the contrast behaviour found in SSFP pulse sequences. A partition represents the magnetisation created, due to excitation by a given rf pulse. In the present model, it is visualised as a set of parameters rather than a vector sum taken over a collection of spins. A multidimensional parameter space, where each dimension is associated with one of the partition parameters, is introduced in order to describe the interaction between partitions and pulse sequence events (e.g., rf pulses and gradients). The three k-space dimensions form the first three dimensions and higher orders are used to handle phase dispersions due to diffusion and main field inhomogeneities. The model makes it possible to perform fast simulation of images resulting from general SSFP pulse sequences. A computer implementation generates images (256 matrix), containing more than 10 different T1/T2 combinations, in less than 45 s on a 120 MHz Pentium computer. The contrast behaviour and signal intensities found in simulated images show excellent agreement with data generated using a clinical MRI scanner system. PMID- 9223047 TI - Ex-vivo MR imaging of liver intracellular contrast agents. AB - The objective is to evaluate whether an ex-vivo model can be used to test intracellular contrast agents for MR imaging of the liver. T1 weighted inversion recovery, proton density spin echo and T2* weighted gradient echo images of the liver were acquired at 0.5 T in 10 rats before and 30 min after intravenous injection of 0.075 mmol/kg Gadolinium benzyloxypropionictetraacetate (Gd-BOPTA, n = 5) or 0.015 mmol/kg dextran magnetite (DM, n = 5), Four additional animals served as controls. After exsanguination and perfusion with saline and formalin, specimens of the liver and brain were embedded in an agar gel and examined with MR imaging one to three weeks later using the same protocol. In-vivo, the mean liver signal enhancement caused by Gd-BOPTA in T1, proton density and T2* weighted images was +23%, +28% and -70%, respectively. The mean liver signal enhancement caused by DM was -71%, -76% and -94%. In-vitro, no signal change was seen in the brain of animals injected with Gd-BOPTA and DM as compared to controls. Liver signal was increased by Gd-BOPTA and decreased by DM. Mean liver enhancement rate induced by Gd-BOPTA was +22%, +5% and +27% for T1, proton density and T2* weighted images, respectively. Mean liver enhancement rate induced by DM was -27%, -19% and -31%. MR imaging signal changes induced by liver intracellular contrast agents are still appreciable in an ex-vivo model. The latter might be useful for for preliminary investigation of intracellular contrast agents for MR imaging of the liver. PMID- 9223048 TI - Correlation of seizure frequency with N-acetyl-aspartate levels determined by 1H magnetic resonance spectroscopic imaging. AB - Using 1H MRSI, we measured N-acetyl-aspartate (NAA), a neuronal marker, in the seizure focus of 16 patients with partial epilepsy. Decreasing NAA correlated with increasing seizure frequency in frontal lobe epilepsy (r = -0.72, p < 0.02) and a similar trend was present in temporal lobe epilepsy (r = -.60, p < 0.06). NAA was not related to the duration of epilepsy. We conclude that patients with higher seizure frequency have evidence of greater neuron loss or dysfunction. PMID- 9223049 TI - NMR measurements of flow profiles in a coarse bed of packed spheres. AB - The velocity profiles of water flowing in a circular pipe filled with coarse, uniform, spherical beads, as well as the evolution of velocity profiles from clear flow into the bed of beads and visa versa, were measured by nuclear magnetic resonance (NMR) imaging using the phase-encoding method. Corrections for partial-volume effects were necessary in order to obtain accurate flow parameters. Phase errors due, for example, to electrical eddy currents induced by switching gradients, were minimized by the use of a static reference sample. For a tube diameter that is eight times the particle diameter, the flow reaches a steady state condition within one bead diameter into the packed bed after having a parabolic velocity profile only 1.5 tube diameters before encountering the bed. Within the packed bed, there are velocity spikes up to 16 times the average velocity in the unobstructed part of the pipe. The velocity distribution in the packed bed has a maximum that approaches the average velocity in the bed for increasing slice thickness along the flow. This distribution decreases exponentially with increasing velocity on the high-velocity side of the maximum. Even at modest flows, where velocity is steady upstream of the packed bed as well as in the bed, unsteady flow is detected downstream of the beads. PMID- 9223051 TI - Direct visualisation of B1 inhomogeneity by flip angle dependency. AB - Inhomogeneities in the spatial distribution of the excitatory Radio Frequency (RF) field, are still a dominant source of artifacts and loss of signal to noise ratio in MR imaging experiments. A number of strategies have been proposed to quantify this distribution. However, in this technical note we present a relatively simple MR imaging procedure which can be used to visualise RF inhomogeneities directly either by means of the magnitude or the phase of an image. To visualise the RF field distribution in both the inner and outer volumes of the coil, we have performed experiments in which the entire coil is submerged in a non-conducting fluid. To the best of our knowledge this strategy has not been used previously in order to evaluate coil performance. Finally, we demonstrate that the method is sensitive enough to reveal the effects of the sample properties on the effective RF wavelength of the transmitted field. PMID- 9223050 TI - Dose distribution of proton beams with NMR measurements of Fricke-agarose gels. AB - Fricke-agarose gels have been irradiated with a proton beam. Then samples have been extracted at different depths with respect to the beam penetration distance, corresponding to different irradiation doses. Relaxation times T1 and T2, measured at 17 MHz, appear sensitive to this kind of radiation. In particular, T2 exhibits three components T2a, T2b and T2c, the first two being sensitive to proton irradiation. At 1% agarose concentration, the relaxation rates R1 = 1/T1, R2a = 1/T2a and R2b = 1/T2b of samples irradiated with both modulated and unmodulated beams, increase with the dose, irrespective of the beam energy. The yield G of Fe3+ ions per 100 eV of absorbed energy is always higher than that obtained for gamma irradiated samples. PMID- 9223052 TI - An improved robust hierarchical registration algorithm. AB - This note describes an improvement to an accurate, robust, and fast registration algorithm (Alexander, M.E. and Somorjai, R.L., Mag. Reson. Imaging, 14:453-468, 1996). A computationally inexpensive preregistration method is proposed, consisting of simply aligning the image centroids, from which estimates of the translation shifts are derived. The method has low sensitivity to noise, and provides starting values of sufficient accuracy for the iterative registration algorithm to allow accurate registration of images that have significant levels of noise and/or large misalignments. Also, it requires a smaller computational effort than the Fourier Phase Matching (FPM) preregistration method used previously. The FPM method provides accurate preregistration for low-noise images, but fails when significant noise is present. For testing the various methods, a 256 x 256 pixel T2*-weighted image was translated, rotated, and scaled to produce large misalignments and occlusion at the image boundaries. The two situations of no noise being present in the images and in which Gaussian noise is added, were tested. After preregistration, the images were registered by applying one or several passes of the iterative algorithm at different levels of preblurring of the input images. Results of using the old and new preregistration methods, as well as no preregistration, are compared for the final accuracy of recovery of registration parameters. In addition, the performances of three robust estimators: Least Median of Squares, Least Trimmed Squares, and Least Winsorized Mean, are compared with those of the nonrobust Least Squares and Woods' methods, and found to converge to correct solutions in cases where the nonrobust methods do not. PMID- 9223054 TI - GABAergic retinocollicular projection in the New World monkey Cebus apella. AB - GABA immunoreactivity was examined in the retina of the New World monkey Cebus apella. Labeled cell bodies were identified as horizontal, bipolar, interplexiform, amacrine and a population of putative ganglion cells. To determine whether ganglion cells were immunoreactive to GABA, double-labeling experiments were performed using Fast Blue as retrograde neuronal tracer injected into the superior colliculus. Retinas containing FB-labeled ganglion cells were subsequently incubated with antiserum against GABA. Although retinocollicular ganglion cells were found in three different layers (ganglion cell layer, inner nuclear layer and inner plexiform layer), our experiments revealed GABA-positive ganglion cells only in the outer half of the ganglion cell layer. PMID- 9223053 TI - Presenilins and early-onset familial Alzheimer's disease. AB - Thirty-seven missense mutations and a splice-site mutation in the presenilin gene PS1 on chromosome 14 and two missense mutations PS2 on chromosome 1 co-segregate with early-onset familial Alzheimer's disease (AD). The presenilins belong to a family of conserved integral membrane proteins which include Caenorhabditis elegans SPE4 and SEL12 and the rat apoptosis-linked gene, ALG3. This review summarizes the genetics of presenilins in AD and indicators of putative function based on cellular localization and the functions of non-human homologues. Findings to date suggest an important role of presenilins in beta-amyloid (A beta) production: in vitro and in vivo studies have shown that presenilin mutations are associated with relatively increased production of the longer, and highly fibrillogenic A beta 42(43) peptide, and a marked elevation in the number of A beta 42-immunoreactive plaques in the brains of individuals with familial AD who carry PS1 and PS2 mutations. There is growing evidence that the deposition of A beta 42(43) could in some cases be an early and key event in the AD pathogenic cascade. The genetic and molecular biological data discussed in this review describe mechanisms by which presenilin mutations could lead to the development of AD. Also, mutant presenilins may be more proapoptotic. It is argued that the understanding of the processes by which presenilin mutations lead to the development of AD will help in devising a coherent framework for therapeutic strategies. PMID- 9223055 TI - IA-type K+ channel blockers promote survival of cortical neurons in culture: involvement of L-type Ca2+ channels. AB - Dendrotoxin (DTX), a well characterized IA-type potassium channel blocker, directly added to the culture medium had no effect on survival of cultured cortical neurons at 6 or 14 days in vitro. On the contrary, neurons exposed to DTX remained in better condition than untreated ones. In an attempt to demonstrate the mechanisms by which DTX may affect neuronal survival we studied its effect in co-cultures of cortical neurons and astrocytes submitted to successive medium changes. After the second change of medium, at 9 days in vitro, the neuronal number in controls decreased by 43%, while in cultures receiving astrocyte-conditioned medium the cell loss was significantly reduced (15%, p < 0.01) with respect to control conditions. When DTX was added to the culture medium neuronal loss was also significantly prevented (25% for 1 microM DTX, p < 0.01) with respect to control conditions. 4-Aminopyridine (4-AP) and 21 mM K+ also preserved neurons. The L-type calcium channel antagonist nifedipine (5 microM) abolished the protective effect of DTX and 4-AP. These results show that K+ channel blockade induces protection against damage produced by repetitive medium change and that this effect is mediated by L-type Ca2+ channels. PMID- 9223056 TI - Developmental surface dyslexia is not associated with deficits in the transient visual system. AB - Deficits of the transient visual system have been reported in unselected groups of dyslexics. The aim of this study was to examine whether this finding holds when subjects with a specific type of developmental reading disorder (surface dyslexia) are considered. Ten Italian children were examined. They all presented the characteristic markers of surface dyslexia: slow and laborious reading with errors in tasks which cannot be solved with a grapheme-phoneme conversion (i.e., homophones). Contrast sensitivity thresholds to phase-reversal gratings were within normal limits for most subjects both for stimuli presented centrally and in the right parafovea. This indicates that developmental surface dyslexia is not associated with a deficit in the transient system. In contrast, sensitivity to high spatial frequency stationary stimuli was reduced. PMID- 9223058 TI - c-fos antisense oligonucleotide prevents delayed induction of hsp70 mRNA after soman-induced seizures. AB - Previous studies have shown the successive expression of c-fos and hsp70 genes, especially in the hippocampal formation, during soman-induced seizures. In order to detect a possible link between the induction of these two genes, antisense strategies have been used. First, the ability of unilateral intrahippocampal infusion of c-fos antisense oligonucleotides to inhibit ipsilateral, seizure related, c-FOS-like immunoreactivity, was verified. Second, induction of hsp70 mRNA was investigated using in situ hybridization. Unilateral inhibition of c-fos induction clearly reduced levels of hsp70 mRNA in the c-fos antisense-infused hippocampus relative to the non-infused contralateral side. Infusion of c-fos sense probe or vehicle did not affect bsp70 mRNA induction. This study suggests a role of c-FOS in regulating bsp70 mRNA induction. PMID- 9223057 TI - Plant-derived amino acids increase hippocampal BDNF, NGF, c-fos and hsp70 mRNAs. AB - Early alterations in mRNAs encoding neurotrophins and stress proteins were investigated following intracerebroventricular injections of beta-N-oxalylamino-L alanine (BOAA) and beta-N-methylamino-L-alanine (BMAA) in adult rats using in situ hybridization. Major increases in heat-shock protein 70, c-fos and brain derived neurotrophic factor (BDNF) mRNAs were seen in hippocampus 1 h after BOAA or BMAA injections. Nerve growth factor mRNA was profoundly increased in the dentate gyrus (DG) after both treatments. Four hours after BMAA injections increased hybridization to BDNF mRNA was still seen in hippocampus, in parallel with reduced neurotrophin-3 expression in the DG. These alterations are in accordance with previous findings of BOAA and BMAA as potent glutamate receptor agonists. PMID- 9223059 TI - Neonatal serotonin depletion modifies development but not plasticity in rat barrel cortex. AB - Effects of serotonin depletion (induced by neonatal injection of 5,7 dihydroxytryptamine) upon dimensions of cortical barrels and their metabolic activation, and upon effects of neonatal vibrissectomy sparing row C, were examined in 1-month-old rats. Dimensions of row C barrels, and of [14C]2 deoxyglucose (2-DG) labelling in the cortex obtained after stimulation of the row C vibrissae, were measured. Serotonin depletion did not change dimensions of barrels, but reduced the extent of 2-DG labelling of cortical representation of the row C whiskers by 30%. Vibrissectomy sparing this row resulted in an expansion of the row C barrels and of 2-DG labelling in the barrel cortex that were similar in both control and serotonin-depleted rats. PMID- 9223060 TI - Anterograde transport of endogenous brain-derived neurotrophic factor in hippocampal mossy fibers. AB - Neurotrophic factors such as brain-derived neurotrophic factor (BDNF) are assumed to provide trophic support via a target-derived, retrograde mechanism of action. However, recent studies suggest that neurotrophic factors can act in an autocrine fashion and perhaps even in an anterograde direction similar to neurotransmitters. To further explore this hypothesis, we compared the neuroanatomical pattern of BDNF mRNA and protein in response to electroconvulsive seizures (ECS) or kainic acid-induced seizure activity. Using in situ hybridization, we found that chronic ECS induced BDNF mRNA predominantly in the granule neurons of the dentate gyrus. However, immunohistochemistry with an anti BDNF antibody revealed that ECS increased endogenous BDNF protein in the mossy fibers, which are composed of axons projecting from the granule neurons of the dentate gyrus to the CA3 pyramidal layer of the hippocampus. Kainic acid administration (10 mg/kg, i.p., once) was used to lesion CA3 neurons selectively, as these are a possible retrograde source of BDNF protein in mossy fibers. Three weeks later, a prolonged elevation of BDNF mRNA in granule neurons, but not elsewhere in hippocampus, was accompanied by an increase in BDNF protein in the mossy fibers. These results suggest that BDNF was transcribed and translated in granule neuron cell bodies but transported in an anterograde direction to provide trophic support of CA3 pyramidal neurons. PMID- 9223061 TI - Relation of age and apolipoprotein E to cognitive function in Down syndrome adults. AB - To test the cognitive effects of aging and apolipoprotein E (APOE) in individuals at high risk for Alzheimer's disease (AD), we assessed APOE genotypes and performance on a battery of neuropsychological tests in 41 non-demented, Down syndrome (DS) adults. Old DS subjects (ages 41-61 years) showed poorer memory and orientation scores than young DS adults (ages 22-38 years), but the groups did not differ in other measures after we controlled for intellectual function. Language ability was inversely related to APOE genotype, even after age was controlled for, with the presence of the epsilon 2 allele corresponding to better language skills than epsilon 4. Age-related cognitive changes in non-demented DS adults are consistent with the early effects of AD. The relationship between basic linguistic skills and APOE genotype supports this genetic factor in influencing the development of dementia and AD neuropathology in DS. PMID- 9223062 TI - The effects of hyper- and hypocarbia on intraparenchymal arterioles in rat brain slices. AB - This investigation examined the direct effects of hyper- and hypocarbia on intracerebral resistance vessels within an intact neuronal synctium. Hippocampal rat brain slices were superfused with artificial cerebrospinal fluid (aCSF). Arterioles were located and diameter changes in response to alterations in aCSF carbon dioxide tension (pCO2) were monitored with videomicroscopy. Microvessels dose dependently dilated and constricted during hyper- and hypocarbia, respectively. A two-fold rise in pCO2 produced a 20% increase in diameter, while a 47% decrease in pCO2 vasoconstricted microvessels by 11%. This is the first model allowing the investigation of the direct actions of physiologic mediators on discrete intracerebral resistance vessels in situ. The results suggest that intracerebral microvessels significantly respond to changes in pCO2 and may be intimately involved in alterations in cerebral vascular resistance. PMID- 9223063 TI - Frontal dysfunction blocks the therapeutic effect of THA on attention in Alzheimer's disease. AB - We evaluated the effect of a single dose of a cholinesterase inhibitor, tetrahydroaminoacridine (THA; 25 and 50 mg, orally), on attention in patients with Alzheimer's disease (AD). THA 50 mg improved performance in attentional measures (Trail Making Test, Big/Little Circle, Simple and Choice Reaction Time) in nine of 28 patients with AD. We analysed retention of 99mTc-labelled ethylene dicysteinate (ECD) in the cortical areas using single photon emission computed tomography. Those patients who benefited from THA treatment had bilaterally higher frontal and prefrontal ECD retention values. We suggest that THA may improve attention in patients with AD, but a severe frontal dysfunction may block the therapeutic effect of THA. PMID- 9223064 TI - Bax accelerates staurosporine-induced but suppresses nigericin-induced neuronal cell death. AB - Bax, a member of the Bcl-2 multigene family, is known to promote apoptosis. To investigate the role of Bax in an experimentally induced cell death of the murine dopaminergic neuronal cell line (MN9D), we established MN9D cells stably over expressing murine Bax (MN9D/ Bax) or vector alone (MN9D/Neo). In MN9D/Neo cells treated with either 1 microM staurosporine or 0.1 microM nigericin, a ladder pattern of DNA fragmentation was induced. As expected, over-expression of Bax in MN9D cells accelerated staurosporine-induced cell death as measured by the MTT reduction assay (62.3% survival in MN9D/Neo vs 27.0% survival in MN9D/Bax). Surprizingly, both nigericin-induced cell death and its accompanying DNA fragmentation were largely attenuated in MN9D/Bax cells (22.0% survival in MN9D/Neo vs 86.7% survival in MN9D/Bax). Similar patterns were observed in two other MN9D/Bax cell lines. Cleavage of poly(ADP-ribose)polymerase caused by nigericin was greatly attenuated in MN9D/Bax cells suggesting that, like Bcl-2, Bax suppresses nigericin-induced cell death by inhibiting the activation of cysteine proteases. Thus, our data imply that Bax acts as a negative or positive regulator of cell death depending on the type of death stimulus applied to the cell. PMID- 9223065 TI - Optical imaging of low Mg(2+)-induced spontaneous epileptiform activity in combined rat entorhinal cortex-hippocampal slices. AB - A reproducible increase in transmission of infrared light was observed during spontaneous seizure-like events (SLEs) induced by low Mg2+ solutions in combined rat entorhinal cortex-hippocampal slices. Comparison of half maxima of transmission change in different regions indicated propagation of SLEs from the medial entorhinal cortex towards the temporal cortex suggesting spread along existing anatomical pathways. Thus, optical imaging of spontaneous epileptiform activity is possible and may improve the assessment of spread patterns. The optical signal outlasted both SLEs and associated K+ signals. In contrast, the tetraethylammonium signal, indicating changes of the extracellular space (ECS) volume, had a longer time course than the transmission changes. ECS volume changes are widely held to be responsible for transmission change. Our data suggest that other mechanisms may also contribute to increased light transmission during epileptiform activity. PMID- 9223066 TI - Carbachol inhibits basal and forskolin-evoked adult rat striatal acetylcholine release. AB - Acutely dissociated adult rat striatal cholinergic neurons labeled with [3H]choline were used in a perifusion system to study muscarinic regulation of basal and forskolin-stimulated fractional [3H]acetylcholine ([3H]-ACh) efflux in the absence of synaptic modulation. Carbachol inhibited basal (40% maximal inhibition; IC50 approximately 0.7 microM) and forskolin-evoked release (75% inhibition; IC50 approximately 0.05 microM) in a concentration-dependent manner, and both carbachol actions were abolished with atropine. Thus, activation of striatal muscarinic cholinergic autoreceptors potently inhibits basal and adenylate cyclase-stimulated ACh release. Tonic inhibitory control of cholinergic activity by functional striatal circuitry apparently prevents detection of these important physiological interactions in slices or in situ. PMID- 9223067 TI - Stability of cortical self-regulation in epilepsy patients. AB - Biofeedback-supported self-regulation of slow cortical potentials (SCP) is increasingly being used for treatment of intractable epilepsy. However, it is unknown whether the acquired ability to regulate one's own cortical potentials remains stable over time. In this study, 18 patients with drug-resistant partial epilepsy performed 35 training sessions in which they learned to generate slow cortical potential shifts in either positive or negative direction. At the end of training, they differentiated significantly between required cortical positivity and required cortical negativity. Six months after this point, they still demonstrated an unchanged between-condition differentiation. The performance in the booster session was particularly good in trials without continuous SCP feedback. The ability to generate positive SCP shifts was related to decrease of seizure frequency during the 6 months follow-up period compared with the 3 month baseline period. This data indicate that the acquired ability of humans to regulate their cortical potentials did not decrease over a 6 month period but rather, tended to consolidate. PMID- 9223068 TI - Glucocorticoids modulate the cellular disposition of lipocortin 1 in the rat brain in vivo and in vitro. AB - The role of glucocorticoids in the regulation of lipocortin 1 (LC1) mRNA/protein expression in the brain is uncertain. This study has examined the influence of dexamethasone on the disposition of LC1 protein in various central and peripheral tissues in the rat. LC1 was readily detectable in all tissues studied by Western blot analysis. Exposure to dexamethasone in vitro (0.1 microM, 3 h) or in vivo (20 micrograms/100 g i.p., 24 h before autopsy) had no discernible effects on intracellular LC1 levels but increased the amount of LC1 attached to the outer surface of cells (pericellular LC1) in cortex, hippocampus, hypothalamus, pituitary gland and peritoneal macrophages (in vitro only). The results suggest that in central tissues, as in the periphery, glucocorticoids promote the translocation of LC1 from intracellular to pericellular sites. PMID- 9223070 TI - Nicotinamide attenuates methamphetamine-induced striatal dopamine depletion in rats. AB - The aim of the present study was to examine the effects of nicotinamide, a co factor in the electron transport chain, on the relationship between methamphetamine (MA)-induced striatal dopamine (DA) depletion and energy metabolism change. Four injections of MA (10 mg/kg, i.p.) at 2 h intervals resulted in decreases of 51% and 23%, respectively, in striatal DA and adenosine 5'-triphosphate (ATP) levels 5 days later. Nicotinamide (500 mg/kg, i.p.) treatment prior to each MA injection attenuated the reductions of striatal DA and ATP contents. Nicotinamide had no long-term effects on striatal DA and ATP levels. These findings suggest that energy impairment might play a role in MA induced DAergic neurotoxicity in the striatum. PMID- 9223069 TI - Distribution of Ca(2+)-permeable AMPA receptors among cultured rat cerebellar granule cells. AB - Some AMPA receptors are permeable to Ca2+. It has been suggested that cultured rat cerebellar granule cells express Ca(2+)-permeable AMPA receptors, but their distribution at a single cell level is unknown. We report that AMPA (in the presence of cyclothiazide) induced Ca2+ entry (measured by Mn2+ quench of fura-2 fluorescence) and intracellular Ca2+ increases in cerebellar granule cells in the absence of extracellular Na+, supporting the presence of Ca(2+)-permeable AMPA receptors. Analysis of intracellular Ca2+ signals in single cells demonstrated a heterogeneous distribution of Ca(2+)-permeable AMPA receptors. PMID- 9223071 TI - Segregated populations of mitral/tufted cells in the accessory olfactory bulb. AB - The dendritic distribution of mitral/tufted (M/T) cells in the opossum accessory olfactory bulb (AOB) was investigated using intracellular injection of Lucifer yellow and DiI labeling. Lucifer yellow labeling demonstrated that the primary dendrites of M/T cells are restricted to one of the two subregions (anterior or posterior) of the glomerular layer. When DiI was placed in the anterior or posterior subregion of the glomerular layer, virtually all of the labeled cell bodies in the AOB were located in the anterior or posterior part of the M/T cell layer, respectively. These results demonstrate that the anterior and posterior subregions of the AOB glomerular layer are termination sites for dendrites belonging to distinct populations of M/T cells. PMID- 9223072 TI - Temporal and spatial dissociation of expression patterns between Zif268 and c-Fos in rat inferior olive during vestibular compensation. AB - We examined the expression of the inducible transcription factors Zif268 and c Fos in the inferior olive during the behavioural recovery following unilateral labyrinthectomy known as vestibular compensation. These transcription factors were differentially induced in the two subnuclei of the inferior olive. In the beta nucleus, c-Fos induction was restricted to the side contralateral to the lesion whereas Zif268 was induced on both the ipsilateral and contralateral side. In the dorsal cap, both proteins were expressed mostly on the contralateral side. In addition to this spatial difference, the expression of Zif268 attenuated faster than that of c-Fos. The present data suggest a functional difference between these two subnuclei of the inferior olive at the level of gene expression in vestibular compensation. PMID- 9223073 TI - Mycobacterium tuberculosis enhances iNOS mRNA expression and HIV replication in human astrocytoma cells. AB - The aim of this study was to investigate whether Mycobacterium tuberculosis (MTB) infection affects human immunodeficiency virus (HIV) replication in T67 human astrocytoma cells and whether HIV and MTB infections induce iNOS mRNA expression in T67 cells. T67 cells were susceptible to both HIVLai and MTB infections, and MTB was able to increase HIV infection in T67 cells, as demonstrated by a marked increase in p24 release. Furthermore, both HIV and MTB infections strongly induced inducible nitric oxide synthase (iNOS) mRNA expression, as verified by RT PCR. These findings suggest that HIV and MTB-induced iNOS expression of astroglial cells may be involved in the neuronal damage associated with HIV infection, particularly in the presence of opportunistic infections such as tuberculosis. PMID- 9223074 TI - Chronic ethanol retards kindling of hippocampal area CA3. AB - To investigate the effects of chronic alcohol which persist long after cessation of ethanol exposure, electrical kindling of the hippocampus was studied in male Sprague-Dawley rats chronically exposed to ethanol. Kindling was initiated 14 days after either one continuous period of ethanol exposure, repeated periods of exposure and withdrawal, or control handling. Animals receiving ethanol exposure required significantly more stimulations to develop behavioral signs of kindling and to attain final kindling criterion than did ethanol-naive controls. Neither the blood ethanol level at the time of withdrawal nor the behavioral signs of acute withdrawal severity correlated with kindling measures. This study indicates that chronic ethanol exposure results in brain alterations which may be assessed long after the cessation of ethanol exposure. PMID- 9223075 TI - Neural cell adhesion molecules play a role in rat memory formation in appetitive as well as aversive tasks. AB - A polyclonal antibody (R1), raised against chick synaptic membrane glycoproteins and recognizing the neural cell adhesion molecule (NCAM) caused amnesia for avoidance tasks when injected into day-old chicks and adult rats 5.5 h post training. We investigated the effects of R1 antibody on memory formation in a non aversive task, where stress is minimal: a massed trial odour discrimination task in rats. Preimmune serum or R1 antibody was injected i.c.v. 5.5 h after the last training session. Forty-eight hours after the training session, control rats showed very good retention whereas R1 antibody injection significantly disrupted retention. The results suggest that glycoproteins recognized by R1 in the rat play a specific role in memory formation for appetitive events as well as in memory formation for aversive situations. PMID- 9223076 TI - Endogenous opioid receptor-like receptor in human neuroblastoma SK-N-SH cells: activation of inhibitory G protein and homologous desensitization. AB - Endogenous expression of opioid receptor-like receptor (ORL1), in human neuroblastoma SK-N-SH cells was demonstrated by binding with nociceptin/ orphanin FQ (N/OFQ). Scatchard analysis of [3H]N/ OFQ saturation binding data gave Kd = 1.3 +/- 0.1 nM and Bmax = 1.58 +/- 2.5 fmol/mg protein. N/OFQ stimulation increased [35S]GTP gamma S binding to cell membranes and attenuated forskolin induced cAMP accumulation in a concentration-dependent manner. The effects of N/OFQ were eliminated by the pretreatment of pertussis toxin (PTX) but not by the antagonists of opioid receptors, revealing mediation of N/OFQ signal transduction by ORL1 receptor and PTX sensitive G protein(s). The ability of N/OFQ to inhibit cAMP production was greatly reduced after prechallenging with N/OFQ, indicating that ORL1 undergoes homologous desensitization in neuronal cells. PMID- 9223077 TI - A specific deficit of dorsal stream function in Williams' syndrome. AB - Williams' syndrome (WS) is a rare, genetically based disorder of cognitive development. Affected individuals show a severe deficit of spatial cognition but a relative sparing of language and face recognition. To examine the possible neural basis of the spatial deficit, we tested a group of WS children, aged 4-14 years, on two measures specific to dorsal cortical stream function: global motion coherence thresholds, in comparison with an analogous form-coherence test, and visuo-manual accuracy in posting a card through a slot, compared with matching the slot orientation. Deficits in these tasks provide the first evidence of specific involvement in WS of the dorsal stream, the cortical system believed to encode information about spatial relationships and the visual control of action. PMID- 9223078 TI - Impaired spatial information processing in aged monkeys with preserved recognition memory. AB - Spatial information processing was examined in a non-human primate model of cognitive aging, using procedures formally similar to tasks designed for rats. The test apparatus was a large open field containing eight reward locations. Monkeys rapidly learned to visit each location once per trial, and probe manipulations confirmed that young animals navigated according to the distribution of cues surrounding the maze. In contrast, aged monkeys solved the task using a response sequencing strategy, independent of extramaze spatial information. Object recognition memory was normal in the aged group. The results reveal substantial correspondence in the cognitive effects of aging across rat and primate models, and they establish appropriate procedures for testing the long-standing proposal that the role of the hippocampus in normal spatial learning is similarly conserved. PMID- 9223079 TI - Anandamide suppresses nitric oxide and TNF-alpha responses to Theiler's virus or endotoxin in astrocytes. AB - Astrocytes are an important cell population in the CNS, involved in cytokine homeostasis and participating in a variety of important physiological and pathological processes. In the present study we showed that primary cultures of neonatal mouse cortical astrocytes stimulated with lipopolysaccharide (Balb/c mice strain, LPS: 1 microgram/ml, 18 h) or Theiler's virus, TMEV (SJL/J mice strain, TMEV: 10(5) PFU/well, 24 h) released an increased amount of nitrites (NO2 ) and tumour necrosis factor-alpha (TNF-alpha) into the culture medium. Exogenous cannabinoids are known to modulate the function of immune cells. Anandamide, an endogenous ligand for the cannabinoid receptor, blocked the release of NO2- and TNF-alpha induced by LPS in a dose-dependent manner. In TMEV-stimulated astrocytes anandamide also suppressed, in a dose-related manner, the stimulatory effects of TMEV on both NO2- and TNF-alpha. It is suggested that anandamide exerts an immunoregulatory role in the CNS. These results could have important implications in the modulation of immunological and inflammatory processes by cannabinoid agents. PMID- 9223080 TI - Endogenous NGF and CNTF levels in human peripheral nerve injury. AB - Nerve growth factor (NGF) is trophic to sensory and sympathetic fibres, and ciliary neurotrophic factor (CNTF) to motoneurones, in animal models of peripheral nerve injury: NGF excess produces hyperalgesia. In this first study of injured human nerves and sensory ganglia, we quantified and localized endogenous NGF and CNTF in 59 neonate and adult patients with brachial plexus and peripheral nerve injury. NGF levels were generally depleted in injured nerves, but relatively preserved acutely in nerve segments distal to injury. NGF immunostaining was observed in Schwann cells in distal nerve segments with pockets of high levels in some neuromas. CNTF levels and immunostaining in Schwann cells were markedly decreased distally within days of injury. We propose that early local administration of NGF and CNTF-like agents may help prevent degenerative changes in injured nerves, while at later stages local anti-NGF treatment (e.g. of some neuromas) may ameliorate chronic pain. PMID- 9223081 TI - Visual motion activates V5 in dyslexics. AB - A recent functional magnetic resonance imaging (fMRI) study concluded that the motion-specific visual area V5 is not activated in dyslexic subjects. We report here opposing evidence based on whole-scalp neuromagnetic recordings. Apparent motion stimuli elicited similar activation of V5 in both dyslexic and control subjects, with a trend for longer latencies in dyslexics. Both high- and low contrast stimuli activated the V5 region in dyslexics. The lack of significant blood flow changes despite modified neuronal synchrony would explain the absence of fMRI signals and the presence of neuromagnetic signals in dyslexic subjects. PMID- 9223082 TI - Early brain potentials link repetition blindness, priming and novelty detection. AB - To study mechanisms of visual object identification in humans, event-related potentials (ERPs) were recorded during successful or unsuccessful identification of rapid, serially presented words (unrepeated or repeated). We observed 'repetition blindness' (RB): more repeated than unrepeated words were incorrectly reported. ERPs from repetition-blinded words exhibited little or none of the enhanced positivity found for correctly reported repeated words, resembling instead ERPs from any unrepeated sequence initially, but only incorrectly reported unrepeated sequences later. Thus it appears that in RB an early (220 ms) neural operation that normally initiates facilitated processing from immediate repetition priming erroneously processes a repeated item as novel. This operation (possibly in basotemporal neocortex) appears to induce differential subsequent processing of novel vs repeated information. PMID- 9223083 TI - Glutamatergic deafferentation of olfactory bulb modulates the expression of mGluR1a mRNA. AB - Glutamate (Glu) released by olfactory nerve axons acts on postsynaptic ionotropic and metabotropic glutamate receptors expressed by principal neurones and interneurones of the olfactory bulb (OB). Using ZnSO4 lesioning of the rat olfactory mucosa and semiquantitative RT-PCR, we examined the effect of removal of the glutamatergic input to the OB on the expression of mGluR1a, mGluR1b and GluR1 mRNAs. Two days after lesioning, mGluR1a mRNA levels in OB increased by 45%. At this time, the expression of tyrosine hydroxylase (TH) mRNA, which is strictly dependent on olfactory nerve input, was still unchanged. In contrast, 16 days after lesioning, deafferented OB exhibited a decrease in both mGluR1a (-30%) and TH (-40%) mRNAs. GluR1 and mGluR1b mRNA levels were not affected at either time point. These results suggest that alterations in glutamatergic input to OB selectively modulate the expression of the mGluR1 splicing form possessing a longer C-terminal domain. PMID- 9223084 TI - IL-1 beta enhances neurite regeneration from transected-nerve terminals of adult rat DRG. AB - We clarified the roles of IL-1 beta in peripheral neural regeneration after axotomy in a three-dimensional collagen gel culture system ranging from a single neurone to a dorsal root ganglion (DRG) explant with its associated nerve bundles. Application of 30 U/ml IL-1 beta to the culture systems clearly enhanced neural regeneration. This regeneration was evident in transected nerve terminals of DRG explants with or without associated nerve bundles, but not in dissociated single neurones. Neural survival was not affected by IL-1 beta in any of these culture systems. These results suggest that IL-1 beta stimulates surrounding non neuronal cells to secrete neurotrophic factors, thus enhancing neurite regeneration from transected nerve terminals in cultured adult DRG explants. PMID- 9223085 TI - Perivascular astrocytes within Alzheimer's disease plaques. AB - The association of astrocytes with plaques is a well-established feature of Alzheimer's disease (AD) and has generally been interpreted as a secondary reaction to amyloid deposition or neuronal degeneration. Astrocytes in brain tissue from six non-demented controls and six patients with AD were investigated using enhanced immunohistochemistry for glial fibrillary acid protein (GFAP) in serial sections from cortex, basal forebrain, amygdala, putamen and diencephalon. Astrocytes colocalized with all diffuse and non-diffuse plaques in AD and control brain tissue. All plaque-associated astrocytes contacted microvessels, and despite having greater numbers of hypertrophic and fine calibre processes, the cells maintained the perivascular arrangement characteristic of control brain tissue. These observations suggest that plaques form at the site of microvascular aberrations followed by reactive and degenerative changes in perivascular astrocytes. PMID- 9223086 TI - Spinally delivered nociceptin/orphanin FQ reduces flinching behaviour in the rat formalin test. AB - The aims of this study were to investigate the dose-dependent effects of spinally delivered nociceptin (0.3, 1, 3.3 and 10 nmol) on flinching behaviour in the rat formalin test and whether these effects were influenced by the concomitant systemic administration of naloxone (3 mg/kg, i.p.). The effect of the highest nociceptin dose differed statistically from vehicle, 0.3 and 1 nmol nociceptin. Following the administration of 1, 3.3 or 10 nmol nociceptin mean total flinches decreased dose-dependently. The effects of 10 nmol nociceptin were not reversed by a high dose of naloxone. We observed a decrease in flinching behaviour with intrathecally to the lumbar enlargement delivered nociceptin and conclude that nociceptin has antinociceptive effects in the rat formalin test. PMID- 9223087 TI - Temporal window of integration revealed by MMN to sound omission. AB - The central auditory system for event perception involves the integrating mechanism of sequential information addressed by the present study. The mismatch negativity (MMN) component of the event-related potentials (ERP) reflects the automatic detection of sound change. ERPs to occasionally omitted stimuli were measured when sequences with constant stimulus onset asynchronies (SOAs) were presented. In separate blocks, the SOA was from 100 to 350 ms. A clear MMN was elicited by a stimulus omission in a sequence of regularly spaced tone pips only when the SOA was shorter than 150 ms, yielding an estimate for the duration of the temporal window of integration used the perceptual segregation of auditory events. PMID- 9223088 TI - Recombinant AMPA receptors with low Ca2+ permeability increase intracellular Ca2+ in HEK 293 cells. AB - Although AMPA receptor subunit configuration controls the Ca2+ permeability of the ion channel, not much is known about Ca2+ signals generated by different AMPA receptor subtypes. We examined the Ca2+ signaling properties of recombinant AMPA receptors using patch clamp to determine ionic permeability properties and Ca2+ imaging to examine changes in intra-cellular Ca2+ level ([Ca2+]i). Activation of recombinant AMPA receptors robustly increased [Ca2+]i even in cells expressing heteromeric receptors containing edited GluRB, which harbored receptors with very low average Ca2+ permeability in patch clamp studies. The results suggest that whereas the GluRB subunit controls Ca2+ permeability, Ca2+ signaling mediated by AMPA receptors correlates poorly with the presence of GluRB. PMID- 9223089 TI - Electrophysiological correlates of temporal and spatial information processing. AB - In order to study the neurophysiological correlates of working memory for different types of information, event-related brain potentials (ERPs) were recorded during a visual spatial and visual as well as auditory duration memory tasks. From stimulus onset to 500 ms ERPs were distinguishable by allocation to visual or auditory modality. From 500 to 2000 ms after stimulus onset, the spatial task generated a parieto-occipital focused negative slow wave, while corresponding ERPs of the temporal task showed a negative slow wave with frontolateral focus. From 1200 to 5500 ms a large positivity was found for the auditory temporal task and for good performers of the visual temporal task. The data suggest a distinction of three processing phases: modality-specific encoding, information-specific encoding and retention in conjunction with modality-specific inhibition processes. PMID- 9223090 TI - Functional MRI evidence for subcortical participation in conceptual reasoning skills. AB - Lesions involving the dorsolateral prefrontal lobes may produce deficits on conceptual reasoning (CR) tasks in humans. Such deficits can also occur with subcortical lesions involving the basal ganglia, thalamus, or cerebellum, suggesting a common, yet widespread, neural network supporting this executive function. Here we report the results of a whole brain functional magnetic resonance imaging (fMRI) experiment in healthy volunteers while performing a CR task. Compared to a sensorimotor control condition, the CR task resulted in discrete subcortical activation sites primarily involving the right basal ganglia, right thalamus and left lateral cerebellum. Cortical activation was present in multiple systems, including the dorsolateral prefrontal and inferior frontal/insular areas; posterior parietal, superior extrastriate, and premotor areas; inferior extrastriate and middle temporal regions; and midline pre supplementary motor and anterior cingulate regions. Our findings provide strong evidence that CR is mediated by interacting neural systems involving the cerebral cortex, basal ganglia, thalamus, and cerebellum. PMID- 9223091 TI - Ultrastructural and immunohistological evidence for dendritic-like cells within human choroid plexus epithelium. AB - Ultrasonic examination of human choroid plexus (CP) disclosed the presence of a possibly new type of CP cells displaying many of the features of dendritic cells and apparently expressing HLA Class II antigens. These cells bear long processes, devoid of tight junctions, inserted between CP epithelial cells, and have close contacts with CP basement membrane. They could, therefore, play a key role in immunosurveillance in the central nervous system. PMID- 9223092 TI - Efficacy and side-effects of clozapine not associated with variation in the 5 HT2C receptor. AB - In the present study we tested the hypothesis that individual response to clozapine treatment and/or the occurrence of side-effects may be influenced by genetic variation in the serotonin 5-HT2C receptor. We investigated the frequency of a common Cys23Ser substitution, which is known to alter the pharmacological properties of the protein, in 152 patients treated with clozapine. Presence of the Ser23 variant was previously reported to predict a good response to clozapine. However, our results did not support an association between genetic variation of the 5-HT2C receptor and response to clozapine. This held true whether the patients were subgrouped for sex and length of treatment. Moreover, we found no consistent association with any of the observed side-effects. PMID- 9223093 TI - Turning of nerve growth cones induced by the activation of protein kinase C. AB - Pathfinding by growing nerve processes in the developing nervous system depends on the turning response of the growth cone to extracellular guidance cues. Embryonic spinal neurons were prepared from 1-day-old Xenopus embryos. After 4 h incubation, a repetitive pulse application was used to produce microscopic chemical gradients near the growth cone. A micropipette containing the protein kinase C (PKC) activator 12-O-tetradecanoyl-phorbol 13-acetate (TPA) or phorbol 12,13-dibutyrate, produced a significant growth cone turning response. A micropipette containing adenosine 5'-triphosphate (ATP) also induced growth cone turning towards the pipette tip. H-7, a PKC inhibitor, inhibited the ATP-induced turning response of the growth cone. Our results suggest that the activation of PKC in developing motoneurons may induce the turning response of growth cones. PMID- 9223094 TI - Functional heterogeneity of left inferior frontal cortex as revealed by fMRI. AB - Using functional magnetic resonance imaging (fMRI), we mapped brain activity in six normal volunteers during two silent verbal fluency tasks, one with a phonemic (letter) cue and one with a semantic (category) cue. In comparison with resting state, both tasks activated the anterior triangular portion of the left inferior frontal gyrus (IFG or F3, for third frontal gyrus) and the left thalamus. There were also areas activated in one task but not in the other: the posterior opercular portion of the left IFG for phonemic fluency, and the left retrosplenial region for semantic fluency. Our findings concur with normal psychophysical data and neuropsychological observations to suggest the recruitment of two overlapping but dissociable systems for the two tasks, and demonstrate functional heterogeneity within the left IFG (Broca's area), where the opercular portion is responsible for obtaining access to words through a phonemic/articulatory route. PMID- 9223095 TI - Seizure-induced glutamate release in mature and immature animals: an in vivo microdialysis study. AB - A glutamate biosensor was used to measure extracellular glutamate concentrations in the hippocampus of mature and immature animals. Significant elevations of extracellular glutamate were observed following seizures induced by either kainic acid or pilocarpine. The degree of glutamate increase following seizures was similar in both mature and immature animals. These results suggest that excitotoxicity may play a role in seizure-induced brain damage in the adult brain. In the immature brain, however, no brain damage is seen after seizures, suggesting that glutamate release may not cause as significant excitotoxic damage early in development. PMID- 9223096 TI - Expression of schwannomin in lens and Schwann cells. AB - Neurofibromatosis type 2 (NF2) is an autosomal dominant genetic disorder characterized by the development of bilateral vestibular schwannomas, meningiomas, ependymomas and juvenile lens opacities. The NF2 gene encodes a tumor suppressor protein, schwannomin (or merlin), with sequence homology to erythrocyte band 4.1, talin, ezrin, moesin and radixin. Using an antibody that recognizes the carboxy-terminal epitope of isoform 1 of schwannomin, we looked at its expression in lens and Schwann cells, two cell-types affected by the NF2 phenotype. Schwannomin was detected as an approximately 80 kDa protein in both cytoplasmic and cytoskeleton fractions. Indirect immunofluorescence localized schwannomin to the cytoplasm and was frequently observed in dynamic cellular regions such as leading edges and ruffling membranes. Its level of expression in the lens inversely correlates with the degree of lens cell differentiation suggesting a role for schwannomin in differentiation-specific events. PMID- 9223097 TI - Differential regulation of adenylyl cyclase in fibroblasts from sporadic and familial Alzheimer's disease cases with PS1 and APP mutations. AB - beta-Adrenoceptor- and forskolin-stimulated adenylyl cyclase activities were determined in primary skin fibroblasts established from patients with sporadic Alzheimer's disease (AD) and from individuals with familial APP KM670/671NL, PS1 M146V and PS1 H163Y mutations. Our data showed a significantly decreased beta adrenoceptor-stimulated adenylyl cyclase activity in fibroblasts from sporadic AD compared with age-matched controls (p < 0.001, Student's unpaired t-test). In contrast, both beta-adrenoceptor- and forskolin-stimulated adenylyl cyclase activities were significantly increased in fibroblasts bearing PS1 M146V and PS1 H163Y mutations compared with controls (p < 0.01 and p < 0.05, respectively). No differences were seen between cell lines with and without the Swedish APP KM670/671NL double mutation. We suggest that various gene mutations associated with AD have different consequences for the regulation of adenylyl cyclase signal transduction in this disorder. PMID- 9223098 TI - Dopaminergic modulation of LTP induction in the dentate gyrus of intact brain. AB - The effect of the dopamine system on the induction of long-term potentiation (LTP) in the dentate gyrus was studied in anesthetized rats. A subthreshold tetanic train (seven pulses at 100 Hz) given to the perforant pathway, which usually fails to elicit LTP, potentiated a slope of field excitatory postsynaptic potentiation (fEPSP) measured from the hilus of the dentate gyrus when a precursor for catecholamine, L-3,4-dihydroxyphenylalanine (L-DOPA), was administered orally to rats. The increase in the fEPSP slope persisted for at least 60 min. Intraventricular injection of a specific dopamine D1/D5 agonist, SKF38393, mimicked the effect of L-DOPA, suggesting an involvement of D1/D5 receptors in the induction of dentate gyrus LTP. Consistent with this, intraventricular administration of the D1/D5 antagonist SCH23390 resulted in complete inhibition of LTP induction by a longer tetanus (100 pulses at 100 Hz), which usually elicits a robust LTP. Thus, D1/D5 receptor activation appears to modulate LTP induction in vivo. PMID- 9223099 TI - A single residue within the homeodomain of the Brn-3 POU family transcription factors determines whether they activate or repress the SNAP-25 promoter. AB - The closely related POU family transcription factors Brn-3a and Brn-3b differ in their effect on a number of different neuronally expressed promoters such as that of the gene encoding the synaptic vesicle component SNAP-25. Thus Brn-3a activates these promoters whilst Brn-3b represses both their basal activity and their activation by Brn-3a. We show here that alterations of a single amino acid at position 22 in the POU-homeodomain from the isoleucine found in Brn-3b to the valine found at the equivalent position in Brn-3a converts Brn-3b from a repressor to an activator of the SNAP-25 gene promoter. The converse mutation in Brn-3a abolishes its ability to activate the SNAP-25 gene promoter and allows it to repress the basal activity of the promoter and its activation by wild type Brn 3a. This is the first time that a single amino acid change has been shown to convert an activator of a naturally occurring promoter to a repressor and vice versa. These results are discussed in terms of the critical role of position 22 in the POU homeodomain in the protein-protein interactions of POU proteins. PMID- 9223100 TI - Reproductive senescence predicts cognitive decline in aged female monkeys. AB - The present investigation provide evidences from a non-human primate model that naturally occurring menopause predicts a prominent signature of age-related cognitive decline. Young and aged rhesus monkeys were tested on a delayed response (DR) task known to the sensitive to aging, and reproductive status was evaluated according to menstrual cyclicity and urinary hormone profiles. Peri /postmenopausal monkeys exhibited significant DR impairments relative to either age-matched premenopausal females, or young control subjects. In addition, markers of endocrine decline in the aged animals were selectively correlated with behavioral performance measures that distinguished premenopausal and peri /postmenopausal monkeys. These results document that menopause is coupled to cognitive decline in the monkey, and they establish a valuable primate model for defining the effects of endocrine aging on brain and behavioral function. PMID- 9223101 TI - Cloning/brain localization of mouse glutamylcysteine synthetase heavy chain mRNA. AB - Glutathione (GSH) is considered the primary molecule responsible for peroxide removal from the brain. Inhibition of its rate-limiting synthetic enzyme, glutamylcysteine synthetase (GCS), results in morphological damage to both cortical and nigral neurons in rodents. Here, we report cloning of the catalytic heavy chain GCS mRNA from mouse and its localization in the murine brain. Heavy chain GCS appears to be localized in glial populations in the hippocampus, cerebellum and olfactory bulb, with lower levels of expression in the cortex and substantia nigra. Variations in GCS levels and subsequent GSH synthesis may explain differences in susceptibility to neuropathology associated with oxidative stress noted in these various brain regions. PMID- 9223102 TI - Time-frequency analysis reveals multiple functional components during oddball P300. AB - A time-frequency decomposition was applied to rare target and frequent non-target event-related potentials (ERPs) elicited in an oddball condition to assess whether multiple functional components occur in the P300 latency range. The wavelet transform (WT) was used because it allows capture of simultaneous or partly overlapping components in ERPs without loosing their temporal relationships. The application of a four-octave quadratic B-spline wavelet transform at the level of single-sweep data allowed us to obtain new information and revealed the presence of separate events during P300 development. Several delta, theta, and alpha frequency components in the P300 latency range differed between target and non-target processing. These findings indicate that P300 is composed of multiple functional components and that the WT method is of use for the study of P300 functional correlates more precisely. PMID- 9223103 TI - Glutamate responsiveness enhanced in neurones expressing amyloid precursor protein. AB - Although the neurotoxicity of beta-amyloid peptide is well established, the cellular functions of amyloid precursor protein (APP) remain unclear. Using an adenoviral vector, we introduced cDNA encoding human APP holoprotein into rat hippocampal neurones in culture. Neurones expressing the membrane-bound form of APP showed greater responsiveness to applied glutamate than non-expressing control neurones, as revealed by Ca2+ fluorometry. This increased responsiveness was not the result of secreted APP, as confirmed by observations of closely spaced APP-expressing and non-expressing cells in the same culture dish. These data suggest that one function of APP may be the regulation of glutamate receptors in neurones. PMID- 9223104 TI - Brain gene regulation by territorial singing behavior in freely ranging songbirds. AB - To investigate the ecological relevance of brain gene regulation associated with singing behavior in songbirds, we challenged freely ranging song sparrows with conspecific song playbacks within their breeding territories. Males responded by approaching the speaker, searching for an intruder and actively singing. In situ hybridization of brain sections revealed significantly higher expression of the transcriptional regulator ZENK in challenged birds than in unstimulated controls in several auditory structures and song control nuclei. We conclude that singing behavior in the context of territorial defense is associated with gene regulation in brain centers that control song perception and production, and that behaviorally regulated gene expression can be used to investigate brain areas involved in the natural behaviors of freely ranging animals. PMID- 9223105 TI - Reduction of excitation by interleukin-1 beta in rat neocortical slices visualized using infrared-darkfield videomicroscopy. AB - The cytokine interleukin-1 beta (IL-1 beta) is thought to be critically involved in the neuroendocrine and behavioral changes which occur in response to systemic infection. In the present study, we have employed the novel technique of infrared darkfield videomicroscopy to examine the effect of IL-1 beta on the intrinsic optical signal (IOS), an indicator of the spread of neuronal excitation and synaptic transmission in the mammalian central nervous system. Low doses of IL-1 beta delivered exogenously to rat neocortical slices produced a reduction of the area of the column-like IOS evoked by orthodromic stimulation. The effect of IL-1 beta was reversible on washout and not mimicked by heat-inactivated IL-1 beta. These results suggest a possible modulatory role of IL-1 beta on synaptic transmission in the rat neocortex which is probably mediated through an activation of GABAA receptors. PMID- 9223106 TI - Presenilin 1 interaction in the brain with a novel member of the Armadillo family. AB - One approach to understanding the function of presenilin 1 (PS1), is to discover those proteins with which it interacts. Evidence for a function in developmental patterning came from C. elegans, in which a PS homologue was identified by screening for suppressors of a mutation in Notch/lin-12, a gene which specifies cell fate. However, this genetic experiment cannot determine which proteins directly interact with PS1. Therefore, we utilized the two hybrid system and confirmatory co-immunoprecipitations to identify a novel catenin, termed delta catenin, which interacts with PS1 and is principally expressed in brain. The catenins are a gene family related to the Armadillo gene in Drosophila, some of which appear to have dual roles-they are components of cell-cell adherens junctions, and may serve as intermediates in the Wingless (Wg) signaling pathway, which, like Notch/lin-12, is also responsible for a variety of inductive signaling events. In the non-neuronal 293 cell line, PS1 interacted with beta catenin, the family member with the greatest homology to Armadillo. Wg and Notch interactions are mediated by the Dishevelled gene, which may form a signaling complex with PS1 and Wg pathway intermediates to regulate the function of the Notch/lin-12 gene. PMID- 9223107 TI - Genetic control of immune responsiveness, aging and tumor incidence. AB - Age-related alterations of the immune system affect both antibody and cell mediated immune responses, T-cell responses being more severely affected than B cell responses. Within the T-cell population, aging leads to replacement of virgin by memory cells and to accumulation of cells with signal transduction defects. Changes in T-cell subsets and in cytokine production profiles may produce suitable conditions for T-cell-mediated disregulation of antibody responses characterized by the production of low affinity and self-reactive antibodies. Also B-cells exhibit intrinsic defects and natural killer (NK) cell activity a profound loss in old mice. Whether age-related immune disfunctions influence life span and tumor incidence has been examined in mice genetically selected for high or low antibody responsiveness. It has been found that genetic selection of vigorous antibody responses in most cases produces mice with longer life span and lower lymphoma incidence. Moreover, the results of genetic segregation experiments indicate that antibody responsiveness and life span are polygenic traits regulated by a small number of the same or closely linked loci. Mice genetically selected for high or low mitotic responsiveness to PHA exhibit low or high tumor incidence, respectively, but no difference in life span, suggesting that T-cell activity is restricted to immune surveillance of neoplastic transformation. Studies on mice genetically selected for resistance or sensitivity to chemical carcinogenesis have uncovered loci that control both resistance to tumor induction and longevity while have no effects on immunity and disease incidence. Thus, the relative role of the immune system in conditioning the duration and the biological quality of life remains to be determined. PMID- 9223108 TI - Changes in apoptosis of human polymorphonuclear granulocytes with aging. AB - Many alterations with aging occur at the cellular and organic levels in the immune system ultimately leading to a decrease in the immune response. Our aim in the present work was to study apoptosis of polymorphonuclear granulocytes (PMN) with aging under various stimulations since apoptosis might play an important role in several pathologies encountered with aging. The PMN of healthy young (20 25 years) and elderly (65-85 years) subjects were examined after 24 h of sterile culture with and without stimulation. The stimulating agents included: phorbol myristate acetate (PMA), hydrogen peroxide (H2O2), N-formyl-methionyl-leucyl phenylalanine (FMLP), granulocyte-macrophage colony stimulating factor (GM-CSF), reduced glutathione (GSH), lipopolysaccharide (LPS) and interleukin 2 (IL-2). Apoptosis was assessed by traditional staining of the plates, by flow cytometric staining and DNA gel electrophoresis. It was found that without stimulation the susceptibility of PMN to apoptosis was slightly increased with aging. Under various stimulations, such as PMA. H2O2, apoptosis was almost 100%, while the treatment by FMLP, oxLDL and GSH did not change its extent in PMN obtained either from young or elderly subjects. Marked age-related changes were observed in the extent of apoptosis under stimulation with GM-CSF, IL-2 and LPS. These agents were able to significantly prevent apoptosis in PMN of young subjects, while only the GM-CSF was able to slightly modulate it in neutrophils of elderly subjects. From these results, we suggest that changes in apoptosis of PMN with aging could play a role in the increased incidence of certain immune system related pathologies of aging, such as cancer, infections and autoimmune disorders. PMID- 9223109 TI - Susceptibility to apoptosis of T lymphocytes from elderly humans is associated with increased in vivo expression of functional Fas receptors. AB - We recently showed that mature T lymphocytes derived from elderly humans were more susceptible to activation-induced cell death than similar cells from young individuals. Because this excessive apoptosis is unrelated to either the age associated decrease in IL-2 production, a differential Bcl-2 expression or to a modification of the antioxidant pathway, we examined the possibility that the Fas receptor (FasR) is directly implicated in the generation of the unwarranted death signal. We investigated the expression and the function of FasR on T lymphocyte populations from healthy young and elderly individuals. We found that the frequency of FasR+ T cells increases as a function of age. The FasR expressed at the surface of freshly isolated T lymphocytes from elderly donors appear to be fully functional since their ligation by a cytocidal IgM anti-Fas mAb leads to a significant increase in DNA fragmentation in this cell population. Conversely, exposure of T cells derived from aged individuals to an antagonistic anti-FasR mAb partially prevents the age-related increase in apoptotic cell death. The population of FasR+ T lymphocytes is essentially constituted of previously activated CD45RO+ cells and also includes recently activated lymphocytes bearing the CD25 and CD69 activation markers. The accumulation of chronically and recently in vivo activated T-cells with age probably contributes to the amplification of the process of Fas-mediated cell death in T lymphocytes isolated from senescent organisms. PMID- 9223110 TI - Effect of aging on cytokine production in normal and experimental systemic lupus erythematosus afflicted mice. AB - The objective of the this study was to determine the cytokine profile of aging mice and to establish whether changes in cytokine production account for the fact that aging mice develop a milder disease than the young in response to induced experimental systemic lupus erythematosus (SLE). Cytokine secretion was evaluated in groups of BALB/c and C3H.SW mice at different ages between 2 and 24 months. The production of IL-2, IL-4, IL-10, IFN gamma and TNF alpha was determined in supernatants of ConA-stimulated splenocytes and that of IL-1 in the supernatants of LPS-stimulated peritoneal macrophages. A gradual age-related decline was observed in the production of IL-2 and IFN gamma, whereas the levels of IL-4, IL 10, IL-1 and TNF alpha progressively increased with aging, in unimmunized BALB/c and C3H.SW mice. Experimental SLE was induced in 2 and 10 month old C3H.SW mice by immunization with the monoclonal anti-DNA antibody bearing the 16/6 Id. The characteristic cytokine profile following immunization of 2 month old mice was early increased production of TNF alpha and IL-1, followed by a peak of Th1 type cytokines (IL-2, IFN gamma). At a later stage of the disease, a peak of Th2 type cytokines (IL-4, IL-10) was observed that was concomitant with low levels of Th1 cytokines. In contrast, in the 10 month old mice that were immunized with 16/6 Id only a mild increase in all the above cytokines was observed. We suggest that the lower autoantibody production and moderate clinical manifestations in aging mice with experimental SLE are causally related to the decreased production of pro inflammatory cytokines at the initial stages of the disease followed by a lower production of both Th1 and Th2 type cytokines. PMID- 9223111 TI - Premature thymic involution, observed at the ultrastructural level, in two lineages of human-SOD-1 transgenic mice. AB - The human Cu/Zn superoxide dismutase (hSOD-1) gene, catalyses the dismutation of O2 to H2O2 and O2. It is located on chromosome 21 in q22.1 and is overexpressed in Down's syndrome (DS) patients. These patients present various abnormalities including mental retardation, congenital heart disease, immunological deficits and premature aging. In order to explore the potential role of SOD-1 overexpression in DS, we have generated two lineages of transgenic mice for the hSOD-1 gene and studied, at the ultrastructural level, the effect of hSOD-1 overexpression on the thymic microenvironment. Modification of the cellular architecture and morphology associated with a lipidic invasion, signs of a premature involution of the thymus, were observed in both lineages. A rupture of the filamentous network in the extracellular and probably also in the intracellular matrix was first observed. These results correlate the thymic alterations visualized in light microscopy, on the thymus from DS patients, and raise the question of the relationship between the SOD-1 overexpression and the different morphological alterations associated with the premature thymic involution observed in SOD-1 transgenic mice. They suggest that thymic and immunological impairments present in DS patients may be related to the SOD-1 gene dosage effect. PMID- 9223112 TI - Effects of the aged microenvironment on CD4+ T cell maturation. AB - Contributing to the functional alterations of the aged immune system is the accumulation of memory CD4+ T lymphocytes and decline in the proportion of naive cells occurring with advancing age. During attempts to alter the naive to memory ratio of CD4+ cells in aged mice, it was observed that regeneration of the peripheral T cell compartment resulted in a population which possessed the same memory cell-enriched characteristics as the unmanipulated age-matched controls. Thymopoiesis in aged mice does not appear to be altered in such a way as to give rise to emigrants with 'memory-like' characteristics. The aged peripheral microenvironment does, however, cause the accelerated maturation of mature, naive CD4+ T cells to the memory state. PMID- 9223113 TI - Senescence and cytokines modulate the NK cell expression. AB - We have been investigating senescence-related changes in human peripheral blood natural killer (NK2) cells. Data accumulated so far consistently and clearly show that both basal and cytokine (IL-2 and interferon alpha and gamma) induced antitumor MHC-unrestricted cytotoxic activity of NK cells are well-preserved in the healthy elderly. To investigate if the non-cytotoxic functions of NK cells are also spared from the influence of senescence, recombinant IL-2-inducible secretion of IFN-gamma, which serves as a first line of defense, was examined. The amount of IFN-gamma secreted by purified, 18 h activated NK cells from the elderly was only 25 percent of that released by the cells from the young. Thus, the type 1 cytokine-inducible cytotoxic and cytokine secretory functions appear to be dissociable properties of NK cells, at least in the elderly. However, this aging-related early phase secretory deficit could be overcome by chronic stimulation with IL-2 (7 day culture). Since different subsets could perform different functions, we analyzed the NK subsets by flow cytometry. A minor CD56bright subset and a major CD56dim subset could be distinguished based on the density of expression of the cell surface CD56 molecule (N-CAM). We inquired if immunosenescence is likely to impact the steady-state level of circulating NK subsets. A significant decrease (P < 0.01) in percent CD56bright among CD56+ cells was observed in the elderly with a relative sparing of the CD56dim subset. The CD56bright/CD56dim ratio, perhaps representing NK cell maturity status, declined with age. This maturation-related subset redistribution and partial loss during aging of high affinity IL-2 alpha beta gamma receptor bearing 'immature' CD56bright NK cells has not been reported before. It could be a consequence of the decline in the level of IL-2 during aging. It is concluded that post adolescent age-associated modulations in human NK cells are not expressed uniformly; they are pronounced in some, subtle in others but negligible in yet other biological parameters. PMID- 9223114 TI - Combination therapy with BRMs in cancer and infectious diseases. AB - In recent years many studies have stressed the importance of using biological response modifiers (BRMs) in the treatment of different conditions of immune impairment correlated with ageing, cancer and infectious diseases. In particular, the use of different BRMs in conjunction with conventional therapies has been extensively explored. Our studies have demonstrated that treatment with Thymosin alpha-1 and low doses of IFN or IL-2 exert powerful biological effects both in vitro and in vivo. They are highly effective in restoring cytotoxic activities in immunosuppression induced by tumors and/or cytostatic drugs. In addition, when combined with specific chemotherapy, they are able to induce a dramatic inhibition of tumor growth in both experimental models and in humans. Immunotherapeutic treatment also has an application in controlling infectious diseases, especially those occurring in the immuno-compromised host. The advantage of using the combined immunotherapy treatment with antiviral drugs has been recently demonstrated by our group both in a murine experimental influenza model and in patients infected with HBV, HCV and HIV. PMID- 9223115 TI - Differential immunologic modulatory effects of tetrahydrocannabinol as a function of age. AB - Tetrahydrocannabinol (THC) the major psychoactive component of marijuana, differentially modulates the immune response of lymphoid cells from young and old mice. This effect was previously shown when cells from mice of different ages were treated with THC in vitro. In the present study THC was given in vivo to mice of different ages. Lymphoid cells from the young and old mice had different immunologic potential in terms of ability to produce cytokines following stimulation with either Con A or anti-CD3 antibody. IL-4 and IL-10 production was consistently up-regulated in spleen cell cultures from the older animals. In addition, in vivo administration of THC up-regulated the proliferative response of lymphoid cells from young adult mice. Such enhancement was not evident for cells from older animals. Although the proliferative response of spleen cells in the old mice tended to be suppressed, statistical significance was not evident possibly because of marked variation of the responses of cells from the older mice. Since marijuana is used by persons of a wide range of ages, aging should be an important variable that must be considered when assessing the immunomodilatory effect of this drug. PMID- 9223116 TI - T lymphocyte proliferative capability to defined stimuli and costimulatory CD28 pathway is not impaired in healthy centenarians. AB - It is generally assumed that T cell proliferation is impaired in aged individuals. We report data on the proliferative capability of peripheral blood mononuclear cells (PBMC) and T lymphocytes from 40 healthy people of different ages, (19-107 years), including 14 centenarians, to defined mitogenic stimuli. We observed no age-related proliferative impairment both in PBMC and in purified T cells stimulated by anti-CD3 mAb or phorbol myristate acetate (PMA). Furthermore, T cells stimulated by anti-CD3 mAb or PMA and costimulated by CD28 mAb did not proliferate differently among young, middle aged subjects and centenarians. Thus, short term T cell proliferation is not affected even at extreme age when well defined stimuli are used on cells deriving from carefully selected healthy subjects. PMID- 9223117 TI - HIV infection and aging: mechanisms to explain the accelerated rate of progression in the older patient. AB - Age is an important predictor of progression in HIV infections. Not only do older individuals' develop AIDS more rapidly than younger persons, they die more quickly after developing an AIDS-defining illness. While the elderly have higher morbidity and mortality rates from viral and bacterial infections, the mechanism(s) responsible for the more rapid progression of HIV infection in older individuals has not been described. Our results demonstrate that the destruction of T cells in both young and old HIV infected patients progresses at the same rate. HIV 1-infected cells from older individuals do not appear more susceptible to immune mediated destruction. The more rapid progression appears due to an inability of older persons to replace functional T cells that are being destroyed. These findings suggest that improved survival in older HIV infected individuals will require more aggressive antiretroviral therapies as well as continued research to identify and preserve immune system elements that control the virus. PMID- 9223118 TI - Influenza (H1N1)-ISCOMs enhance immune responses and protection in aged mice. AB - Aging is associated with a decline in immune function and the elderly are therefore more susceptible to infectious disease and less responsive to vaccination. Influenza antigens complexed as immunostimulatory complexes (ISCOMs) generate more potent protective immune responses compared with non-adjuvanted flu antigens in young adult mice. We report on the protective efficacy of flu-ISCOMs compared with the current split flu vaccine in an aged mouse model. DBA/2 mice aged 2 or 18 months were immunized with flu vaccine, ISCOMs or live virus, prior to challenge with the homologous virus. In aged mice, flu-ISCOMs induced significantly higher serum hemagglutination inhibition (HAI) titers compared to vaccine, similar to the levels obtained in young adult mice that received the split vaccine. Flu-ISCOMs but not vaccine induced cytotoxic T lymphocyte (CTL) responses in young and to a lesser degree in aged mice. In aged mice flu-ISCOMs significantly reduced illness and enhanced recovery from viral infection compared with vaccine. Our data suggests that flu-ISCOMs may offer an improved vaccine strategy for protection of elderly humans against the complications of influenza infection. PMID- 9223119 TI - The aging retarding effect of 'Long-Life CiLi'. AB - 'Long-Life CiLi' ('CiLi') oral liquid, is composed of superoxide dismutase (SOD), polysacchairide, vitamin C, vitamin E and trace elements which were all extracted from a natural plant fruit Cili (Rosa roxburghii Tratt) in Guizhou, China. A set of indices were evaluated after administration of 'CiLi' 10 ml Bid, for two months in 50-75 years old healthy people, the mean value of NK cell activity (22.4 +/- 10.8-->27.5 +/- 12.9%, P < 0.05), SOD (453.0 +/- 24.2-->468.6 +/- 21.3 micrograms/gHb, P < 0.001), Catalase (15.5 +/- 1.7-->17.4 +/- 3.0 U/mgHb, P < 0.001) and GSH content (2.3 +/- 0.3-->2.6 +/- 0.5 mg/gHb, P < 0.001) in erythrocytes and 'delta CO, CI, SV, SI, LVET, LVETI and AC' values increased significantly, while the serum LPO level (4.20 +/- 0.78-->3.78 +/- 0.50 nmol/ml, P < 0.001), total microcirculation weighed value (1.87 +/- 1.0-->0.92 +/- 0.5, P < 0.001), delta PVR (-241.7 +/- 733.2-->187.9 +/- 938.2, P < 0.05) and the light reaction time (Simple RT, red light: 383 +/- 128-->332 +/- 68.9 ms, P < 0.05; selective RT: red light 709 +/- 287-->566 +/- 119 ms, P < 0.05; green light 639 +/- 162-->536 +/- 80 ms, P < 0.01) decreased significantly. There were no significant differences in the control group. The mean life span of fruit flies were significantly elongated for low, medium and high concentrations 'CiLi' treatment groups than in the control group (Female: 57.6 +/- 11.3-->62.1 +/- 12.8; 69.6 +/- 14.7; 62.6 +/- 12 days; P < 0.05 approximately 0.001. Male: 56.3 +/- 9.6-->64.9 +/- 12.4; 64.5 +/- 14.5; 64.8 +/- 14.1 days, P < 0.001). It is suggested that 'CiLi' has an aging retarding and geroprotection effect. PMID- 9223120 TI - Age-related changes in T cell surface markers: a longitudinal analysis in genetically heterogeneous mice. AB - In a longitudinal design in which each animal was examined repeatedly throughout its life, we used flow cytometry to determine the numbers or proportions of a variety of T lymphocyte subsets in the peripheral blood of mice aged 8-20 months. Half of the 128 mice were given a low calorie diet which is known to extend lifespan and to reduce the rate of age-associated change in T cell immune function. In these genetically heterogeneous mice, bred as the progeny of CB6F1 females and C3D2F1 males, aging led to statistically significant increases in the number of CD3 cells and the proportions of CD4 memory and CD8 memory cells, and declines in the proportions of CD4 cells and CD4 virgin cells. Older mice also had increased proportions of CD4 and CD8 cells that were able to extrude the fluorochrome R123 through the action of P-glycoprotein. Caloric restriction delayed or prevented most of these age-dependent changes, but had little effect on expression of P-glycoprotein. Correlation analysis showed that those individual mice with high levels of CD4 memory or CD8 memory T cells tended also to have relatively low levels of CD4 virgin cells and low proportions of CD4 T cells, particularly at older ages. In addition, those mice which had the greatest rate of increase in CD4 memory and CD8 memory T cells also tended to show the greatest decline in CD4 virgin and in the proportion of CD4 cells. High numbers of CD4R (and high rates of change in the CD4R subset) were associated with high numbers (and change scores) for the CD8R subset, but there were no strong correlations between the R123-extruding subsets and other age-sensitive markers. Thus some, but not all, age-sensitive T cell subsets show correlated levels and correlated rates of change throughout the middle of the lifespan. PMID- 9223121 TI - Enzymatic defenses of the rat heart against lipid peroxidation. AB - Normal ageing is associated with different changes in the cardiovascular system that lead to an increase in pathological processes such as hypertension and heart failure. Therefore the importance of glutathione peroxidase and catalase for protection against peroxidation was studied in the rat heart. Each of the these enzymes was regulated by feeding rats a low selenium diet either unsupplemented or supplemented with 0.4 parts per million of selenium, with or without the catalase inhibitor, sodium fluoride, in their drinking water. After 2 months, selenium deficient rats had 87% reductions in mitochondrial and cytosolic glutathione peroxidase activities. These reductions were accompanied by increased peroxidation in heart homogenates and mitochondrial suspensions. Since increased mitochondrial peroxidation only occurred when both the cytosolic and mitochondrial glutathione peroxidase activities were involved, these selenoenzymes appear to work in tandem and reductions in both are a prerequisite for increased peroxidation in the heart. Peroxidation did not occur in sodium fluoride treated rats even though cytosolic catalase activity was inhibited by 70%. Moreover, inhibition of catalase activity did not exacerbate the level of peroxidation in selenium deficient rats depleted of glutathione peroxidase activity. Because increased peroxidation was only associated with reductions in glutathione peroxidase activity irrespective of catalase activity, the selenoenzyme appears to be more important for detoxification of hydrogen peroxide in the heart. PMID- 9223122 TI - Proton magnetic resonance spectroscopy can differentiate Alzheimer's disease from normal aging. AB - In order to evaluate the pattern of proton magnetic resonance spectroscopy (1H MRS) in the gray and white matter of patients with Alzheimer's disease (AD) and healthy controls, a cross-sectional study was carried out on 13 consecutive AD patients and 7 healthy older subjects who were referred to the Day-Hospital for diagnostic assessment. All examinations were performed on a 1.5 Tesla whole-body scanner. Volumes of interest were selected in both the gray (temporal region) and the white (frontal region) matter. N-acetyl group, total creatine, total choline and myo-inositol were quantified referring the metabolite peak area to the unsuppressed water peak area acquired under the same conditions, and the ratio was expressed in arbitrary units. A significant decrease in N-acetyl-aspartate (NAA) in both gray and white matter and an increase in myo-inositol (mI) in gray matter of AD patients were observed. The gray matter NAA/mI ratio clearly separated the two groups. White matter mI was significantly associated with severity and duration of dementia. No association with age was documented. It can be concluded that in vivo 1H-MRS can contribute to the knowledge of pathophysiology of AD, giving neurochemical details of both gray and white matter. In particular, the gray matter NAA/ml ratio seems to be able to differentiate normal cerebral aging from Alzheimer's disease. PMID- 9223124 TI - Electrophysiological evidence of an increase in cold tolerance of cardiac muscles in mice after energy restriction. AB - Life-prolonging energy restriction (ER) has been known to extend longevity. The heart was selected as the target organ of ER and the electrophysiological properties of ER on the heart were investigated. Action potential parameters were measured on ventricular papillary muscles of C57BL/6 mice (2-6 months of age). Resting membrane potential (Rm) did not change even when the temperature was lowered to 20 degrees C in ER mice (-67.5 +/- 0.8 mV), however, the membrane was depolarized in the control (-61.1 +/- 1.1 mV). Action potential duration measured at 30 and 50% repolarization was significantly prolonged in ER mice at 20-35 degrees C. Ouabain (10 microM) decreased Rm in ER mice at 20 degrees C (-68.6 +/- 1.0 to -63.6 +/- 0.8 mV), but failed to decrease Rm in the control (-60.6 +/- 1.8 to -62.1 +/- 1.2 mV). There were no significant differences in extracted Na, K ATPase activity or affinity and binding capacity of ouabain between ER and control hearts. These results indicate that in ER mice the lack of effect of temperature on Rm was not due to a change in the physicochemical properties of Na, K-ATPase. The present study collectively suggests that ER increases cold tolerance in the heart of mice. PMID- 9223123 TI - P53 synthesis and phosphorylation in the aging diet-restricted rat following retinoic acid administration. AB - Multiple doses of retinoic acid (RA) were administered intraperitoneally to three groups of male Fischer 344 rats over a 36 h period. The p53 isoforms from bone marrow nuclei in these three groups of rats were analyzed over time by two dimensional polyacrylamide gel electrophoresis (PAGE) and fluorography for the incorporation of [35S]methionine (p53-synthesis) and [32P]phosphate (p53 phosphorylation). Two groups of rats, young (3.5 months) ad libitum (Y/AL) and old (28 months) ad libitum (O/AL), had free access to Purina rat chow; a third group of old (28 months) diet-restricted rats (O/DR) were maintained on a restricted caloric intake (60% of the AL diet) from 3 months of age. After 36 h of RA dosing, the PAGE patterns of p53 synthesis and phosphorylation in Y/AL and O/DR rats were very similar. In both groups, an increase in complexity was observed with labeling of additional isotypes possessing more acidic isoelectric values. In contrast, the O/AL animals showed a pattern of p53 isoform synthesis and phosphorylation that was considerably less complex and lacked the pronounced shift to more acidic forms following RA dosing. The p53 isoforms of O/AL rats as recognized by wild type (wt) Pab 246 antibody, were also much less dramatic in their increase to more acidic forms. Two-dimensional phospho-tryptic maps of Y/AL and O/DR rats were also very similar, both exhibiting two additional minor 32P labeled fragments after RA dosing. The maps of O/AL rats did not show the two additional fragments following RA administration. After RA dosing, cyclin protein inhibitors (p16, p21, p27) revealed robust labeling with their respective antibodies in Y/AL and O/DR rats as analyzed by Western blotting. The O/AL animals showed marginally detectable antibody recognition of the cyclin inhibitors after RA dosing. Taken together, these data suggest that the biosynthesis and phosphorylation of p53 isoforms and the expression of cyclin dependent kinase inhibitor proteins is not significantly different between Y/AL and O/DR rats. Further, these results confirm and extend our previous observations that chronic diet-restriction attenuates the age related decline in the metabolic activity of nuclear protein products. PMID- 9223125 TI - cAMP signalling mechanisms with aging in the Ceratitis capitata brain. AB - Aging has been associated with alterations in protein phosphorylation. This study was undertaken to examine eventual changes in cAMP-dependent protein kinase (PKA) activity and enzyme regulatory subunit levels from the dipterous Ceratitis capitata brain with postmaturational aging and senescence. PKA activity was determined in cytosolic and membrane fractions of the C. capitata brain during the adult stage of the insect lifespan. PKA activity markedly increased at the first stages of the life of the fly both in cytosol and in membranes. A lower peak of PKA activity was evident both in particulate and cytosolic fractions in the terminal phase of the life of the fly. Thus, PKA activity was significantly higher in the brain of mature flies when compared to the brain of aged flies. It is possible that increases in cAMP-dependent protein phosphorylation levels characterize the terminal aging process in the insect nervous tissue. On the other hand, levels of regulatory (R) subunit were also measured in membranes and cytosol by immunoblotting. Cytosolic regulatory subunit levels were more elevated near the terminal phase of life, whereas in membranes, regulatory subunit levels decrease in senescence in parallel with particulate PKA activity. The increased R subunit level in cytosol may reflect a cellular response mechanisms to down regulate the kinase system in aged flies. PMID- 9223126 TI - Human biological decline and mortality rates. AB - Intercepts on the x (age)-axis of 107 normalized declining human biological functions were determined and assembled in 3 histograms, being placed in increasing order within each decade (10 year period). The histograms were classed accordingly as they contained properties associated with dividing cells, sensory properties and non-dividing cells respectively. Their cumulants were determined, multiple regressions calculated and compared with current death-rates for women and men respectively, for 10 amongst the longest living populations in the World. An alternative verification based on risk theory led to an estimate of an optimal life expectancy of 96 years. The survival curve turns out to be of the form (See text: Formula) where the inner integral represents the cumulant dimension (t') and the outer one age (t"). The premises underlying this study are compatible with the notion of a probable life-span, rather than a fixed one. PMID- 9223127 TI - The effect of donor age on human erythrocyte density distribution. AB - Erythrocyte density has been reported to increase with in vivo aging. Gradient centrifugation techniques allow different density erythrocyte fractions to be purified from whole blood. As the donor age increases there is a concomitant increase in the percentile contribution of whole blood by a light density fraction. Similarly, a heavy density component of erythrocytes decreases with increasing donor age. In the present study these changes were investigated and found to exhibit a logarithmic relationship to donor age. The etiology of this is unknown but may be due to a decreased erythrocyte lifespan in older individuals. PMID- 9223128 TI - First Alfred Meyer Memorial Lecture. Epileptic brain damage: a consequence and a cause of seizures. AB - Alfred Meyer and his colleagues were the first to report (1954-1956) that the most frequent pathology in tissue from patients with complex partial seizures successfully treated by anterior temporal lobectomy is mesial temporal sclerosis, and that the majority of patients with this lesion give a history of a prolonged seizure early in life. These observations have been repeatedly confirmed. Experimental data from animal models strongly supports the hypothesis that a prolonged generalized or limbic seizure in early life damages the hippocampus and other limbic structures, facilitating an epileptogenic process that, after a latent period, gives rise to spontaneous limbic seizures. Some mechanisms potentially contributing to this process have been identified. PMID- 9223129 TI - DNA fragmentation in normal development of the human central nervous system: a morphological study during corticogenesis. AB - Refinement of the cell number by programmed cell death is a major morphogenetic mechanism of the developing central nervous system (CNS) in vertebrates including mammals, which determines to a significant degree its mature cytoarchitecture. We have examined the topography and the extent of cell death in different regions of the human CNS prenatally (11 fetuses), and in the early post-natal weeks (three newborns). Attention was focused on the wall of the telencephalon during a relatively short time period (12th-23rd week of gestation), corresponding to the time of major proliferation in the ventricular zone and to the peak of neuronal migration; both these mechanisms are crucial for corticogenesis. The TUNEL method was used, allowing the recognition of cell death because of its ability to label blunt ends of double-stranded DNA breaks. Morphological features of nuclei at different stages of apoptosis were identified, providing better evidence of the extent of the process than histological stains. Cell labelling was seen in either post-mitotic elements in the ventricular zone, or along the migratory pathways in the intermediate zone and subplate at all prenatal ages examined. No apoptotic nuclei were seen in the cortical plate. These findings suggest that apoptotic cell death drives the selection of cells which are committed to play a role during the early stages of corticogenesis. Lack of evidence of clonally related apoptotic cells also indicates that cell death occurs randomly. Therefore, molecular signals from the surrounding microenvironment seem to be necessary for the apoptotic pathway to be turned on, thus determining the fate of post-mitotic cells. PMID- 9223130 TI - Viliuisk encephalomyelitis--review of the spectrum of pathological changes. AB - Viliuisk encephalomyelitis (VE) is an unique neurological disease occurring in the Iakut (Sakha) people of Siberia. Evolution of the disease follows one of three broad clinical forms: subacute, slowly progressive or chronic. Death occurs within 3 to 6 months in subacute cases and within 6 years in the slowly progressive cases. Chronic cases lack a subacute phase but show a slowly progressive dementia associated with bradykinesia, dysarthria and spastic paraparesis that stabilizes late in the disease process. In subacute and slowly progressive cases, focal necrotizing encephalomyelitis is seen at necropsy. Chronic cases show multifocal areas of lysis with a gliotic margin, predominantly within grey matter, lacking associated chronic inflammatory changes seen in the other forms of the disease. Epidemiological studies are consistent with a disease of low-grade communicability, but laboratory studies have so far failed to reveal an infectious organism. The spectrum of neuropathological changes are reviewed in this examination of 11 cases. Although the aetiology of VE remains obscure, further studies are warranted since it may represent a novel disease process. PMID- 9223131 TI - Apoptosis in measles virus-infected human central nervous system tissues. AB - The extent of apoptotic cell death was examined in central nervous system (CNS) tissues from three cases of subacute sclerosing panencephalitis (SSPE). Apoptosis was demonstrated by in situ end-labelling of DNA in formalin-fixed, paraffin embedded tissue sections. Measles virus and cell types were labelled by immunohistochemistry and/or in situ hybridization. Furthermore, bcl-2 expression in SSPE was examined by immunohistochemistry. All three cases exhibited varying degrees of apoptosis in all CNS areas studied. Brain tissue from a non neurological control case did not show any significant apoptosis. Characterization of cell types demonstrated neurons, oligodendrocytes, lymphocytes and microglia undergoing apoptosis. A linear relationship could not be established between virus burden and the extent of apoptosis in any particular area. Virus-negative cells were observed which were undergoing apoptosis. Bcl-2 immunoreactivity in SSPE was confined to the infiltrating cell population. These results suggest that apoptosis of various cell types may contribute to the neuropathogenesis of measles virus infection in the human CNS, either as a direct effect of viral infection or by cytokine-mediated responses. PMID- 9223132 TI - The role of the polyamine inhibitor eflornithine in the neuropathogenesis of experimental murine African trypanosomiasis. AB - The treatment of late-stage human African trypanosomiasis is complicated by a post-treatment reactive encephalopathy, also referred to as a 'reactive arsenical encephalopathy', that may be fatal. This study used a well established experimental mouse system to assess the use of the trypanostatic drug, eflornithine, in the management of this post-treatment reaction. Female CD-1 mice infected with an eflornithine-resistant trypanosome stabilate and treated with the trypanocidal compound diminazene aceturate on or after day 21 post-infection develop a reactive encephalopathy and relapsing parasitaemia. If these animals are re-treated with diminazene aceturate, a severe encephalopathy develops histologically comparable with that of human cases and characterized by a severe meningoencephalitis and astrogliosis. Histopathological and immunocytochemical examination shows that administration of eflornithine before or after the development of this reactive encephalopathy prevented or ameliorated the inflammatory reaction. Since an eflornithine resistant stabilate was used, this effect appears to be independent of the drug's trypanostatic action and illustrates an important, previously unrecognized, pharmacological property of eflornithine. Consideration can now be given to the use of eflornithine for the management of human trypanosomiasis cases, even where trypanosome resistance to eflornithine exists. PMID- 9223133 TI - Long-term effects of Semliki Forest virus infection in the mouse central nervous system. AB - Semliki Forest virus (SFV) infection of mice is used as a model to study pathogenic processes occurring in viral encephalitis and demyelinating disease. In this study, the long-term effects of infection by the avirulent M9 mutant of SPV on the central nervous system (CNS) of BALB/c and SJL mice were determined. The presence of infectious virus, viral RNA and cytokine mRNA in the brains of individual mice and the presence of lesions in the spinal cords of the same mice up to 360 days post-infection (d.p.i.) were analysed in order to detect any correlation between these parameters of pathogenesis. Infectious virus could not be detected beyond 7 d.p.i. for either mouse strain. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the presence of the E2 and nsP1 regions of the virus genome and mRNA for interferon-gamma and tumour necrosis factor-alpha. Viral RNA could be detected up to 90 d.p.i. for both mouse strains. Cytokine mRNA could be detected up to 28 d.p.i. for BALB/c mice but up to 360 d.p.i. for SJL mice. Inflammatory lesions, which were associated with cytokine mRNA expression, were not detected in BALB/c mice beyond 28 d.p.i. but were detected in two SJL mice at 90 d.p.i. It is concluded that M9-SFV infection induces long-term prolonged expression of pro-inflammatory cytokines in the CNS of the majority of SJL (but not BALB/c) mice which is not associated with persistence of the virus genome. M9-SFV infection of SJL mice may be a relevant model for the pathogenesis of multiple sclerosis in man. PMID- 9223135 TI - Region-selective glutamine synthetase expression in the rat central nervous system following portocaval anastomosis. AB - Glutamine synthetase (GS) content was investigated using immunohistochemical methods in the hippocampus, cerebellum and spinal cord of rats after long-term portocaval anastomosis (PCA). Six months after surgery, GS content was increased in several areas of each region and decreased in others, compared with controls. In the hippocampus, the CA1-CA3 pyramidal subfields and the dentate molecular layer had a high level of GS expression; PCA reduced GS content in other hippocampal regions, such as the dentate hilus. In the cerebellum, PCA significantly increased GS immunoreactivity in the Bergmann glial processes of the molecular layer and decreased GS immunoreactivity in astrocytes of the granule cell layer. In the spinal cord, GS immunoreactivity increased in the dorsal horn and decreased in the ventral horn. Blood vessels located in zones with GS-immunopositive perineuronal astrocytes in PCA-exposed brains were surrounded by strongly GS-immunostained perivascular processes. These results suggest that PCA exposure had a differential effect on GS expression in different regions of the central nervous system. The increased immunoreactivity of GS positive cells in PCA-exposed brains correlates with glutamatergic areas, which may contribute to protecting neurons against extracellular glutamate and/or ammonia excess. PMID- 9223134 TI - Expression of pro- and anti-apoptosis gene products in brains from paediatric patients with HIV-1 encephalitis. AB - We have previously demonstrated the presence of DNA fragmentation in neurons, macrophages and microglia consistent with apoptosis, but not in reactive astrocytes in brain tissue from paediatric patients with HIV-1 encephalitis (HIVE). To further understand the underlying mechanism(s) for these findings as they relate to gene-directed neural cell death, we studied the in-situ expression of the Bcl-2 family of proteins, including the pro-apoptosis gene product Bax, the anti-apoptosis gene product Bcl-2, and Bcl-x. We demonstrate significantly elevated numbers of Bax-positive microglia and macrophages immunoreactive in basal ganglia and cerebral cortex of children who had HIVE, in comparison to HIV 1 infected children without encephalitis or children who were seronegative for HIV-1. In contrast, patients with HIVE, but not HIV-1 without encephalitis, or seronegative controls, had increased expression of Bcl-2 and Bcl-x in reactive astrocytes in cortex and basal ganglia. In vitro studies using Western blot analysis demonstrated an up-regulation in the levels of Bax, and phosphorylated (i.e. inactive) Bcl-2 in HIV-1 infected macrophages, and in LPS-activated macrophages, relative to levels in virus-negative unstimulated macrophages. These results suggest that productive HIV-1 infection, or cellular activation, renders macrophages more vulnerable to apoptosis. Taken together, these findings suggest that brain-resident macrophages and microglia in patients with HIV-1 encephalitis are more prone to undergo apoptosis and that astrocytes in contrast may be resistant to apoptosis. This may represent a mechanism to limit microglial activation and the spread of productive HIV-1 infection in the CNS of children with HIV-1 encephalitis. PMID- 9223136 TI - Diagnostic (clinical and morphological) criteria for adult neuronal ceroid lipofuscinosis (Kufs' disease), Hopital de la Salpetriere 'AFM Institut de Myologie', Paris, France, 5 December 1996. PMID- 9223137 TI - The human prion diseases--from neuropathology to pathobiology and molecular genetics: an update, Vienna, Austria, 6-7 December 1996. PMID- 9223138 TI - Illusory memories in amnesic patients: conceptual and perceptual false recognition. AB - Little is known about the neuropsychology of false recognition. D.L. Schacter, M. Verfaellie, and D. Pradere (1996) induced false recognition in amnesic patients and normal controls by exposing them to numerous semantic associates of a nonstudied word and found that amnesics showed significantly reduced levels of false recognition. To determine whether this outcome is specific to the semantic domain, the authors examined false recognition after exposure to lists of conceptually and perceptually related words. In the control group, conceptual false recognition was associated with "remember" responses and perceptual false recognition was associated with "know" responses. Amnesic patients showed reduced levels of conceptual and perceptual false recognition that were approximately equally divided between remember and know responses. PMID- 9223139 TI - Rates of forgetting in organic amnesia following temporal lobe, diencephalic, or frontal lobe lesions. AB - Forgetting rates were examined in patients with diencephalic, temporal lobe, or frontal lesions. No significant differences were found in short-term forgetting of verbal and nonverbal material; in recognition memory for pictures, words, or designs over delays between 1 min and 20 or 30 min; or on a measure of explicit cued recall for words, calculated in terms of the process dissociation procedure. Significantly faster forgetting was found in the diencephalic and the temporal lobe groups in the free recall of pictures of objects, although there was no difference between these 2 groups. It is concluded that the major deficit in amnesic patients' memory processes is in the initial acquisition of information but that there is a subtler deficit in retention over specific delays, detectable only on measures of free recall. PMID- 9223140 TI - Memory impairment in multiple sclerosis: a quantitative review. AB - To assess the nature and magnitude of memory impairment in multiple sclerosis (MS), the authors analyzed quantitatively 36 studies comparing the memory performance of MS participants to healthy controls. The authors studied (a) the pattern of impairment across short-term memory (STM), working memory (WM), and long-term memory (LTM); (b) the moderating influence of retrieval support on LTM impairment; (c) the covariation of WM and LTM impairment; and (d) the moderating influence of clinical characteristics of the MS sample on memory impairment. The analyses revealed significant impairment across all memory domains and failed to support a retrieval-based account of LTM dysfunction in MS patients. In addition, robust associations were found between clinical features of MS and memory impairment. The findings suggest a more global pattern of memory deficits in MS than has been previously believed, with deficits clearly associated with neurological disability and disease course. PMID- 9223142 TI - Generation effects and source memory in healthy older adults and in adults with dementia of the Alzheimer type. AB - Recognition and source memory were explored in healthy older adults, adults diagnosed with very mild dementia of the Alzheimer type (DAT), and adults diagnosed with mild DAT. Two sentence-completion tasks were used. In Task 1, half of the sentences were completed (clozed) by the participant, and half by the experimenter. In Task 2, half were participant clozed, and half were participant read (already clozed). Recognition of the cloze words and accuracy of categorizing them as participant generated or experimenter generated (Task 1) and participant generated or participant read (Task 2) were measured (source discrimination). Contrary to previous reports, the DAT groups showed the generation effect, that is, better recognition for participant-generated words than experimenter-generated words (Task 1) or read words (Task 2). Source discrimination was disproportionately impaired in the DAT groups. PMID- 9223141 TI - Effects of aging on conditional associative learning: process analyses and comparison with focal frontal lesions. AB - Conditional associative learning (CAL), a measure validated in studies of frontal lesions, was used to evaluate the hypothesis that age-related cognitive decline is related to frontal dysfunction. Older adults and focal frontal participants showed impaired CAL performance, but the deficit was greater in the latter group, where it was specific to participants with dorsolateral prefrontal cortical (DLPFC) lesions. The deficits were attributable to strategic rather than basic associative processes. Error scores described ways in which past information failed to guide behavior, and they were related to lesion location. Congruence between older adults and DLPFC participants on a measure of defective inhibition suggests that age-related decline in inhibitory processes is due to DLPFC dysfunction. PMID- 9223143 TI - Effects of relatedness and number of distractors on attribute judgments in Alzheimer's disease. AB - Participants made judgments about the relative salience of category exemplars (e.g., fruit: apple or grape) or parts (e.g., plane: wings or seats). Mildly affected Alzheimer's disease (AD) patients were as accurate but slower than normal controls, and their response times increased more for related (e.g., apple, grape, or fig) than unrelated (e.g., apple, gym, bandit) choices as the number of alternatives was increased from 2 to 3. Performance (accuracy and response times) of moderate-severely affected patients was poorer still, but number of distractors and relatedness did not interact. In combination with previous findings (e.g., M. K. Johnson, A. M. Hermann, & J. L. Bonilla, 1995), these results suggest that the reflective processes necessary for deciding among competing alternatives show disruption early in the disease process. Such processing deficits would compound any difficulties arising from a degrading semantic structure. PMID- 9223144 TI - Impairment in category fluency in ischemic vascular dementia. AB - The underlying mechanisms for impaired output on letter (F, A, and S) and category (e.g., animal) word list generation (WLG) tasks in subcortical ischemic vascular dementia (IVD) were investigated. Normal control (NC) and Alzheimer's disease (AD) participants were also studied. IVD and NC participants performed better on category than letter WLG tasks, whereas the opposite was observed among AD participants. IVD participants produced fewer responses than AD participants on letter WLG tasks, but there was no difference between AD and IVD participants on the "animal" WLG task. AD participants scored lower than IVD and NC participants on animal WLG indexes measuring semantic knowledge. There were few differences between IVD and NC participants. The reduced output on the animal WLG task for IVD participants is consistent with search-retrieval deficits. The reduced output of AD participants may be caused by degraded semantic knowledge. PMID- 9223145 TI - Cognitive predictors of incident Alzheimer's disease: a prospective longitudinal study. AB - The present study examined whether cognitive variables measured at baseline could predict incident cases of Alzheimer's disease (AD) after a 3-year follow-up period. Twenty-six incident AD adults and 179 very old (M = 83.5 years) adults without dementia participated in a population-based study. Cognitive performance was indexed by the Mini-Mental State Examination (MMSE) and multiple indices of memory and visuospatial and verbal performance. A logistic regression analysis that controlled for age, gender, and education indicated that MMSE scores were reliable indicators of who would develop AD. In addition, recall of organizable words, recognition of faces, and letter fluency were reliable predictors of subsequent dementia status after differences in MMSE performance were partialed out. Thus, although the MMSE is useful in predicting dementia, there is an additional advantage of assessing specific indices of cognitive functioning. Further, supportive episodic memory tasks may be more salient predictors of incident AD than tasks that offer less supportive encoding or retrieval conditions. PMID- 9223146 TI - Stroop color-word interference and electroencephalogram activation: evidence for age-related decline of the anterior attention system. AB - Groups of healthy, community-dwelling younger and older adults performed a Stroop task in which color and word could be congruent or incongruent and spatially integrated or separated. During the task, continuous electroencephalogram (EEG) was recorded from frontal, parietal, and occipital regions. The magnitude of the Stroop interference effect and task-related EEG activation was greater for older than younger adults when stimuli were integrated. This effect was significant over medial and lateral frontal and parietal, but not occipital, regions. In comparison, interference and EEG activation did not differ for younger and older adults when stimuli were separated. These findings support the hypothesis that the anterior attention system is more sensitive to the effects of increasing age than the posterior attention system. PMID- 9223147 TI - Examination of age-related deficits on the Wisconsin Card Sorting Test. AB - Adult age differences in Wisconsin Card Sorting Test (WCST) measures were examined before and after statistical control of age-related differences in measures of feedback usage, working memory, and perceptual-comparison speed. The proportion of age-related variance associated with a summary measure of WCST performance was greatly reduced after controlling for measures of feedback usage, working memory, and perceptual-comparison speed. Furthermore, the age-related variance associated with the feedback-usage measure was reduced after controlling for working memory and perceptual-comparison speed measures. These results are consistent with the idea that age-related performance differences in the WCST are partially mediated by adult age differences in feedback usage and that age differences in feedback usage are mediated by age differences in working memory, which are in turn-mediated by age-related reductions in processing speed, indexed by measures of perceptual-comparison speed. PMID- 9223148 TI - Is it possible to be schizophrenic yet neuropsychologically normal? AB - This study identified and characterized a group of schizophrenic patients without neuropsychological (NP) impairment. A comprehensive NP battery was administered to 171 schizophrenic outpatients and 63 normal comparison participants. Each participant's NP status was classified through blind clinical ratings by 2 experienced neuropsychologists; 27% of the schizophrenics were classified as NP normal. The NP-normal and NP-impaired schizophrenics were similar in terms of most demographic, psychiatric, and functional characteristics, except that NP normal patients had less negative and extrapyramidal symptoms, were on less anticholinergic medication, socialized more frequently, and were less likely to have had a recent psychiatric hospitalization. The existence of NP-normal schizophrenics suggests that the pathophysiology underlying the cognitive deficits often associated with schizophrenia may be distinct from that causing some of its core psychiatric features. PMID- 9223149 TI - Postconcussion syndrome occurs in children. AB - The consensus of evidence published since 1924 suggests that parents report attention deficits, hyperactivity, or conduct disorder after pediatric head injury rather than postconcussion syndrome. In this study, the symptoms reported by children after mild (n = 38) and moderate-severe (n = 27) head trauma were compared to those reported after orthopedic injury (n = 47) and to adults matched for injury severity and chronicity by using a structured interview based on diagnostic criteria for postconcussion syndrome. Pediatric head trauma caused significantly more subjective symptoms after 6 weeks than orthopedic injury. These symptoms were related to head injury severity and the child's anxiety level. When assessed in a similar manner, children report postconcussion syndrome similar to that seen in adults. PMID- 9223150 TI - Not peripheral parasitaemia but the level of soluble parasite antigen in plasma correlates with vaccine efficacy against Babesia canis. AB - Groups of five dogs were vaccinated against Babesia canis using soluble parasite (SPA) antigens from in vitro cultures. Although vaccination did not significantly alter peripheral parasitaemia upon challenge, protected animals had lower levels of SPA in the plasma after a challenge infection. The severity of anaemia correlated with the SPA-load during the post-challenge period in that high levels of SPA were associated with low haematocrit values. In addition, it was found that recovery was associated with the production of antibodies against SPA. The results suggest that SPA induce anaemia during B. canis infection, and that vaccination with SPA results in antibody production that can neutralize the effects of SPA after a challenge infection. PMID- 9223151 TI - Protection of mice against Schistosoma mansoni infection by passive transfer of sera from infected rabbits. AB - Sera from rabbits infected with unattenuated Schistosoma mansoni cercariae conferred significant levels of protection against S. mansoni challenge (P < 0.001) after passive transfer to mice. Infected rabbit sera were only effective in conferring protection when transferred during the first week of infection, and were not effective when administered against liver-stage worms. Immunoglobulins isolated from the infected rabbit sera with Protein A-Sepharose were shown to be responsible for the transfer of protection to mice. Immunofluorescence studies demonstrated that the sera were more reactive against the surface of three hour old mechanically transformed schistosomula than against the surfaces of lung stage schistosomula. The sera from infected rabbits reacted polyspecifically against antigens in cercaria, schistosomula, and the worm and egg stages of the S. mansoni life-cycle. The host parasite relationship of S. mansoni in the rabbit is discussed. PMID- 9223152 TI - The tegument of Schistosoma mansoni: functional and immunological features. AB - Some structural, biophysical and molecular features of the tegument are shown, highlighting their changes during maturation as important adaptative mechanisms for the survival of the parasite and resistance to immune attack. On the other hand, many antigens, targets of the immune response against the schistosome, are located on the tegument. Some of the features of these molecules are presented and the possibilities of using them in the immunological control of the disease are discussed. PMID- 9223153 TI - Cross-reactive idiotypes on rabbit anti-SEA antibodies stimulate anti-idiotype spleen and lymph node cell responses of mice infected with Schistosoma mansoni. AB - Antibodies to Schistosoma mansoni soluble egg antigens (SEA) generated in outbred rabbits from two different sources were used to study cross-reactive idiotype/anti-idiotype (Id/anti-Id) interactions in S. mansoni-infected mice in two different locations. Immunoaffinity purified rabbit polyclonal anti-SEA antibody preparations (RabId) were predominantly Ig by SDS-PAGE and demonstrated anti-SEA activity by ELISA and Western blot. RabId prepared from three separate rabbits and used at 40 micrograms/ml in vitro stimulated lymphocyte proliferative responses of spleen cells from mice with eight week infections (Mean +/- SEM [E C]) of 3869 +/- 764, 18594 +/- 3046 and 8962 +/- 1734 cpm. Spleen cells from uninfected mice exposed to the same RabId preparations in vitro had respective [E C] values of 206 +/- 144, 494 +/- 154 and 363 +/- 180. Lymph node cells from mice infected for 8 weeks demonstrated [E-C] of 123 +/- 400, 5073 +/- 828 and 2361 +/- 656 upon exposure to these 3 RabId preparations. Lymph node cells from uninfected mice had [E-C] of 220 +/- 76, 194 +/- 82 and 143 +/- 72 when exposed to these RabId. Maximum in vitro proliferative response to Ig from unimmunized rabbits was 761 +/- 400 by spleen cells from mice with eight week infections. These data indicate the presence of cross-reactive Id on rabbit anti-SEA antibodies from different rabbits that can stimulate in vitro lymphoproliferative responses of spleen or lymph node cells from S. mansoni-infected mice. PMID- 9223154 TI - Characterization of a secreted antigen of Onchocerca volvulus with host protective potential. AB - A cDNA designated MOv2 was isolated from an Onchocerca volvulus library on the basis of its product's recognition by an antiserum raised against the infective stage. Immunogold electron microscopy revealed a high density of antigenic sites associated with the annulae of the L3 cuticle and with the uterine wall of the adult female: a general, low density of labelling occurred in all developmental forms. Western blotting confirmed the presence of the antigen throughout the life cycle and the existence of an immunologically cross-reactive homologue in the related filaria, Acanthocheilonema viteae. The antigen was shown to be secreted by infective larvae and adult females of A. viteae. Sequence comparisons revealed two homologues of MOv2 (Ov-20, Ov-9) which had been selected independently by other laboratories on the basis of their specific recognition by human onchocerciasis infection sera. The IgG antibody response against MOv2 in cattle experimentally infected with O. lienalis revealed the induction of a response during the prepatent period that was strongly boosted at the onset of patency. However, only a proportion of infected cattle responded with a detectable level of anti-MOv2 antibodies. The appearance of MOv2 in larval cuticle and secretions prompted us to evaluate it as a candidate molecule for prophylactic immunization. Trials performed in the A. viteae/Mongolian jird model of filariasis revealed that recombinant MOv2 induced a host-protective response, reducing worm recoveries by 36-55% following a challenge infection. PMID- 9223155 TI - Involvement of accessory cells in the Trypanosoma cruzi-induced inhibition of the polyclonal response of T lymphocytes. AB - Infection with Trypanosoma cruzi is characterized by hyporesponsiveness of the immune system during the acute phase of infection. To better understand the immunological mechanisms affected by T. cruzi, we studied if a reduced T cell proliferative response could originate from an inability of T cells to proliferate or a functional deficiency at the level of accessory cells (AC). The inhibitory effect exerted by T. cruzi was during the induction phase of the lymphoproliferative response, suggesting the participation of AC in the hyporesponse. Then we further investigated the potential of the parasite to interfere with accessory cell-dependent and -independent pathways of human T cell proliferation. Peripheral blood mononuclear cells and peripheral blood lymphocytes from healthy individuals, enriched for T cells, were analysed with regard to their proliferative capacity using: phytohaemagglutinin, immobilized anti-CD3 monoclonal antibody (MoAb) and MoAb to the CD28 antigen, anti-CD3 MoAb and recombinant IL-2 and anti-CD3 MoAb plus phorbol myristate acetate in the presence of parasites. Significant suppression of the proliferative response was caused by the parasite only when AC were present. The parasite markedly reduced the surface expression of HLA-DR and CD11b antigens, key molecules in PHA-induced proliferation. Addition of indomethacin to the culture failed to reverse the inhibitory effect of the parasites, suggesting that prostaglandin E2 was not involved. These data suggest that AC in contact with T. cruzi become incompetent as antigen presenting cell because they are unable to induce a normal proliferative response in T lymphocytes. PMID- 9223156 TI - Paramyosin: a candidate vaccine antigen against Schistosoma japonicum. AB - Paramyosin, a 97 kDa myofibrillar protein, is a candidate vaccine antigen for prevention of infection with the human parasite Schistosoma mansoni. To determine if paramyosin would also induce protection against Schistosoma japonicum, paramyosin was biochemically purified from S. japonicum adult worms. SDS-PAGE demonstrated a single protein with a molecular weight of 97 kDa. In four separate experiments, vaccination of mice with S. japonicum paramyosin without adjuvant induced significant resistance (62%-86%, P < 0.001) against cercarial challenge as compared to controls. These data suggest that S. japonicum paramyosin may represent a candidate vaccine for immunization against schistosomiasis japonica. PMID- 9223158 TI - Macrophage modifying factor secreted by the tetrathyridia of Mesocestoides corti (Cestoda): monoclonal antibody to the modifying factor antagonizes its immunological activity. AB - Immunomodulation of macrophage activity by in vitro secretions of Mesocestoides corti has been previously demonstrated. The modifying activity secreted by M. corti had the effect of reducing the normal accessory function of macrophages in a Con-A-activated lymphocyte proliferation assay. This paper describes the purification of the modifying activity by FPLC techniques and the generation of a monoclonal antibody (MoAb) to this molecule in mice. The MoAb bound immunomodulatory FPLC fractions of M. corti in an ELISA. When MoAb was applied in conjunction with immunomodulatory parasite secretions to macrophages in vivo or in vitro, the modifying effect of the secretions was abolished. This profound effect of the MoAb should help to elucidate the mechanisms by which metacestode parasites avoid host immune responses and may enable therapeutic intervention. PMID- 9223157 TI - Evidence that cellular immune responses to soluble and membrane associated antigens are independently regulated during human schistosomiasis mansoni. AB - We have made a comparative analysis of human cellular and antibody responses to membrane associated adult worm antigens (Mb-A), soluble adult worm antigens (SWAP) and soluble egg antigens (SEA) derived from Schistosoma mansoni. Chronically infected patients with the intestinal (I) and hepatosplenic (HS) forms of the disease as well as non-infected putative immune 'endemic normals' (EN), were studied. We observed that the cellular responses, of individuals, to the two adult worm preparations, SWAP and Mb-A, may be distinct and can be related to the occurrence of resistance or pathology. The resistant group (EN) presented higher levels of both cellular proliferation, and IFN-gamma production, in response to Mb-A as compared with SWAP whereas HS individuals presented higher levels of cellular proliferation to SWAP as compared with Mb-A. Individuals with intestinal disease had similar levels of proliferation to both antigens. The response to SEA by all groups was generally similar, and not predictive of any clinical form. The specific antibody response to the three antigens were in general higher among infected patients than in resistant EN individuals. These results support the hypothesis that the response to adult worm antigens may be pivotal in determining both the development of resistance and severity of disease. PMID- 9223159 TI - Synergistic action of cyclosporin A and polyspecific rabbit anti-sera against murine Schistosoma mansoni. AB - The efficacy of cyclosporin A (CsA) treatment against Schistosoma mansoni in mice was compared with treatments that included co-administration of one of two anti sera (infected rabbit serum (IRS) obtained by repeated infection and a worm membrane antigen anti-serum (WSS) obtained by immunization with worm surface supernatants). These two sera recognized a number of worm antigens but differed in precise detail. Administration of CsA alone to mice harbouring mature infections of S. mansoni reduced worm burdens and preferentially targeted female worms. Sera administered alone had no effect on worm burdens. Co-administration of worm membrane antigen anti-serum (WSS) with CsA reduced worm burden significantly compared with drug treatment alone. Male worms were more susceptible to this combined treatment regime. Anti-infection serum (IRS) had a lesser stimulatory activity in combination with CsA which was not statistically different from the effects of CsA alone on worm burdens. The data suggest that CsA-induced surface damage to the parasite may reveal specific antigens that were previously unavailable for host attack. PMID- 9223160 TI - Brugia pahangi in cats: the passive transfer of anti-microfilarial immunity from immune to non-immune cats. AB - Serum from cats (Felis catus) that were repeatedly infected with Brugia pahangi and had become amicrofilaraemic (mf-ve) was injected intravenously into microfilaraemic (mf+ve) cats. If more than 1 microliter of immune serum per 1000 mf was injected, microfilarial counts fell dramatically within minutes and, in some cats, mf completely disappeared. In most cases mf reappeared 21-44 days later. However, in two experiments mf never reappeared and circulating antigen (indicative of the presence of living adults) could not be detected. At autopsy no adult worms were found, but in one cat 6 mf/ml were detected by filtration of cardiac blood. Passive transfer of single Ig isotypes showed that IgG is the immunoglobulin responsible for the mf killing effect of immune serum, and that IgG1 is probably the most active isotype. Mf killing and destruction, occurred in the lungs in an antibody dependent cell mediated reaction involving neutrophils, eosinophils and mononuclear cells. Three of the 20 recipient cats died from what appeared to be anaphylactic shock while under anaesthesia probably due to the sudden release of inflammatory mediators in the lung. PMID- 9223161 TI - In vitro stimulation of naive mouse lymphocytes by Heligmosomoides polygyrus adult worm antigens induces the production of IgG1. AB - The production of high levels of IgG1, by mice chronically infected with the parasitic nematode Heligmosomoides polygyrus, has been documented for a number of years. In order to investigate this phenomenon, naive lymphocytes from B10.D2 mice were incubated in vitro with H. polygyrus adult worm homogenate (AWH) and the culture supernatants examined for immunoglobulin production. Stimulation of pooled naive splenocytes by AWH was found to produce IgG1, but not IgM, in an antigen dose dependent manner. Identical stimulation of splenocytes of individual inbred mice, indicated that this effect was reproducible but with considerable variation between mice. When the IgG1 produced was tested for specificity, it was found that there was little evidence that the immunoglobulin produced was able to bind to the inducing parasite antigens. Analysis of purified T cells reconstituted with splenocytes, demonstrated that T cells were the target lymphocytes of the stimulating molecule, contained within AWH. These results show that H. polygyrus AWH can induce the production of non-parasite specific IgG1 from naive splenocytes and that this production is crucially dependent upon the cell content of the in vitro culture. Furthermore, the production of IgG1 is not proportional to the degree of lymphocyte proliferation. It is suggested that at least part of the hypergammaglobulinaemia produced during a primary H. polygyrus infection, is due to this non-specific stimulation of mouse lymphocytes by the parasite. PMID- 9223162 TI - Structural and molecular specificity of antibody responses in mice immune to third stage larvae of Onchocerca volvulus. AB - Immunization of mice with irradiated Onchocerca volvulus infective stage larvae (L3) has been demonstrated to confer protection against challenge infections with these larvae. Additionally, cytokine level measurements and cytokine depletion studies have shown that both IL-4 and IL-5 are important in generating a protective immune response against O. volvulus challenge infections, thus suggesting a dependency of protective immunity on IgG1, IgE and/or eosinophils. In the present study, we examined the humoral responses of immunized mice to O. volvulus L3 antigens. ELISA measurements of total serum antibody levels indicated that IgE was the only antibody isotype elevated in mice immunized with O. volvulus L3. IgM from immunized mice was the only isotype that recognized surface antigens on intact O. volvulus L3. IgG1, IgG3, IgE and IgA recognized internal parasite antigens on O. volvulus L3 frozen sections. Western blot analysis of L3 proteins showed that in serum from mice immunized with O. volvulus L3 IgG1, IgG2a/2b, IgA, and IgE, as well as IgM, recognized unique L3 proteins. Antibodies in serum from L3 immunized mice were able to detect O. volvulus adult antigens in a pattern similar to the recognition found in O. volvulus L3. Some L3 antigens were shared by adults, while other antigens were L3 specific. The ELISA, immunohistochemistry and Western blot findings thus demonstrate a complex pattern of antigen recognition of parasite antigens by antibodies found in mice immune to the L3 of O. volvulus. PMID- 9223163 TI - Association between anti-Pfs48/45 reactivity and P. falciparum transmission blocking activity in sera from Cameroon. AB - Pfs48/45, a sexual stage parasite protein doublet of P. falciparum, is a target of antibodies which inhibit the development of the parasite in the mosquito. Twenty-eight (54%) out of 52 sera of gametocyte carriers from Cameroon reduced infectivity in the mosquito membrane feeding bioassay to less than 20% of the controls. These 52 sera were analysed by competition ELISAs for the presence of antibodies capable of competing the binding of six monoclonal antibodies (MoAbs) directed against five different epitopes on Pfs48/45. The percentage of these 52 Cameroon sera that competed with one of the MoAbs ranged from 13% (epitope I) to 33% (epitope IIc). Comparison of activity in the transmission-blocking assay (> or = 80%) and in the Pfs48/45 competition ELISA show a relative specificity of 100% (24 of 24) and a relative sensitivity of 75% (21 of 28). Non-blocking sera showed no competition with any of the MoAbs. These MoAbs were further used to study the diversity of epitopes among isolates of P. falciparum using a two-site ELISA. MoAbs against epitope I, III and V reacted with four different isolates whereas epitope II could be subdivided into three epitopes. None of the isolates reacted with MoAb 3G12 (epitope IV). Using these four different isolates, the competition ELISA titre varies from 1/20 to 1/80 and no significant differences are found between the isolates except for epitope II where only three out of 11 positives for epitope IIa were also positive for epitope IIc. PMID- 9223164 TI - Is the induction of apoptosis the mechanism of the protective effects of TNF alpha in helminth infection? AB - Tumour necrosis factor alpha has been implicated in protective immune responses to a number of parasitic helminths. However, the final effector mechanisms resulting in death or expulsion of the parasite are unclear. Here we suggest that, by employing phylogenetically conserved mechanisms, the mammalian immune system is able to interfere with helminth development directly and that the protective effects of TNF alpha in helminth infections may operate via the induction of parasite apoptosis. PMID- 9223166 TI - Stage-specific expression of surface molecules by the larval stages of Haemonchus contortus. AB - Three monoclonal antibodies, Hc2, Hc22 and Hc6, were produced against a surface extract of L3 Haemonchus contortus and screened against both free living and parasitic stages of the parasite. Hc2 and Hc22 were of IgG2c isotype and their target epitopes were insensitive to periodate treatment. Hc6 was of IgM isotype and its reactivity was sensitive to periodate treatment of the antigen, suggesting that Hc6 is specific for a carbohydrate epitope. Western blotting of larval extracts and staining of live worms demonstrated that Hc2 was specific for the surface of second stage larvae and that the epitope was still present on the protective sheath of third stage larvae but absent from the L3 cuticle itself. Hc22 was found to be specific for a 70-90 kDa antigen on the surface of third stage infective larvae and cross reacted with a higher molecular weight species present on the surface of third stage Teladorsagia circumcincta. Hc6 showed strong binding to three high molecular weight proteins on immunoblots of third stage larvae only, but in contrast to Hc2 and Hc22, showed no binding to the surface of live larvae. These studies demonstrate unique expression patterns of stage specific antigens on the surface of free living and parasitic H. contortus larvae. PMID- 9223165 TI - GM-CSF-induced priming of human neutrophils for enhanced phagocytosis and killing of asexual blood stages of Plasmodium falciparum: synergistic effects of GM-CSF and TNF. AB - Granulocyte macrophage-colony stimulation factor (GM-CSF), which is a haematopoietic cytokine generated by activated T lymphocytes and macrophages during infection, was investigated for its effects on human neutrophil-mediated killing of asexual blood forms of Plasmodium falciparum. Pretreatment of neutrophils with human recombinant-GM-CSF markedly increased the parasite killing (measured by a radiometric assay), in the presence of normal serum (containing complement), immune serum (IS), purified IgG (from IS) or heat inactivated IS. GM CSF pretreatment also enhanced phagocytosis of the parasite by neutrophils and the expression of CR3, Fc gamma RII and Fc gamma RIII receptors. Treatment of neutrophils with a combination of GM-CSF and TNF resulted in a synergistic increase in phagocytosis and killing of the parasite. The findings suggest that GM-CSF is likely to form part of the cytokine network responsible for regulating the antiparasitic activity of the neutrophil in malaria. PMID- 9223168 TI - Fasciola hepatica: rapid switching of stage-specific antigen expression after infection. AB - Early developmental stages of the trematode parasite Fasciola hepatica were collected from the peritoneal cavity and liver of mice during a ten day infection period. Using one dimensional SDS-PAGE, differences in protein expression profiles were observed in stages collected on the same day post-infection in different physiological locations and also in juvenile parasites collected from the same location on different days post-infection. Four rat monoclonal antibodies were raised against the parasite using lymph nodes draining infected tissues. Three monoclonal antibodies, FY3-1, FY3-2 and FY4-7, were generated using cells from the mesenteric lymph node of recently challenged immune rats, while FY1-6 was derived from hepatic lymph node cells of a chronically infected rat. The epitope recognized by FY3-2 appeared to be carbohydrate in nature and was present on the surface of newly excysted juveniles. Immunoblots revealed that the antigens recognized by FY3-1, FY3-2 and FY4-7 were only expressed for two days after infection. In contrast, FY1-6 recognized epitopes expressed across all developmental stages screened. The rapid changes in protein and antigen expression observed during the early stages of infection may assist the parasite to evade the host immune response. PMID- 9223167 TI - Evidence for the existence of ganglioside molecules in the antigen of Entamoeba histolytica. AB - Gangliosides were found to be present in Entamoeba histolytica. They were extracted from lyophilized trophozoites of the pathogenic strain HM-1:IMSS and purified by high performance thin-layer chromatography. Two resorcinol-positive bands, comigrating with GM2 and GD1a were demonstrated, revealing the existence of ganglioside molecules in Entamoeba histolytica. The GM2 content, determined as lipid-bound sialic acid, was 1.5 micrograms/10(8) amoebae, the content of the GD1a comigrating band was 0.32 microgram/10(8) amoebae. The identity of the GM2 comigrating band was confirmed by TLC immunostaining, using the monoclonal anti GM2 antibody GMB28. Furthermore, six out of ten anti-amoeba positive sera selectively reacted with the GM2 comigrating band, as revealed by immunostaining on TLC plates. Absorption tests revealed that preincubation of anti-amoeba positive sera with standard GM2 was followed by a significant decrease in the reaction with amoeba trophozoites by indirect immunofluorescence. These results demonstrate that a GM2 comigrating component of Entamoeba histolytica may be one of the antigens responsible for the appearance of circulating antibodies in patients with amoebiasis. PMID- 9223170 TI - Can software be used to predict antigenic regions in Plasmodium falciparum peptides? AB - We compared the antigenicity of p126 Plasmodium falciparum peptides with predicted antigenic regions identified using the methods described by Garnier et al. (1978) and Chou & Fasman (1974). For this purpose nine different P. falciparum peptides were synthesized in accordance with the deduced amino acid sequence of the p126 gene, and their reactivity was tested using an enzyme linked immunosorbent assay against sera from individuals with a natural malaria infection. Both predictive methods gave similar antigenic-index scores, however, a comparison of these predictive results with data obtained by ELISA showed that the probability of a correct prediction was only around 45% for both cases. Thus, in our view computer software could not be used in isolation for screening purposes, and other parameters must also be taken into account when using such software to assess antigenicity. PMID- 9223169 TI - Sm480: a high molecular weight Schistosoma mansoni antigen associated with protective immunity. AB - Rabbit antisera were raised against an antigen present in Schistosoma mansoni adult worms and eggs, and were shown to yield a single immunoprecipitin arc in immunoelectrophoresis and immunodiffusion against S. mansoni egg and worm antigen extracts. The antisera conferred partial but significant protection (22-30%) against a S. mansoni challenge when transferred to mice five and six days after the mice had been infected percutaneously with 200 cercariae. The egg and the worm forms of the antigen were immunologically cross-reactive, but the egg antigen possessed peptidolytic activity that could be inhibited with serine protease inhibitors. In indirect immunofluorescence the rabbit antisera reacted with surfaces of cercariae, five-day old lung-stage schistosomula, miracidia and praziquantel-treated adult worms. Gel-filtration chromatography demonstrated a relative molecular size of approximately 480 kDa for both the egg and worm forms of the antigen, and lectin-affinity chromatography indicated both were glycosylated. The serine protease activity and large relative molecular size of egg Sm480 were confirmed by a combination of radiolabelling with tritiated di isopropyl fluorophosphate, immunoelectrophoresis and polyacrylamide gel electrophoresis. PMID- 9223171 TI - A single gene copy merozoite surface antigen and immune evasion? AB - During the course of chronic malaria infection antigenic variants of a parasite antigen are expressed and exposed on the surface of infected erythrocyte membranes. There also exists a number of apparently invariant single gene copy blood-stage antigens, exposed or non-exposed, which have been shown to afford immunity under experimental conditions. To determine why the host, presented with invariant 'protective' antigens, is unable to control infections effectively, immunity to a representative single gene copy antigen, the merozoite surface protein 1 (MSP1) was investigated in Plasmodium chabaudi chabaudi AS, a murine model of chronic malaria. Immunization with monoclonal antibody affinity purified native MSP1 resulted in enhanced control of parasitaemia on challenge, irrespective of the parasite inoculum size; challenge with a single parasite, however, suggested that expansion of resistant parasite subpopulations was not occurring. Challenge of mice immunized with recombinant fusion proteins encoding N- or C-terminal regions of the P.c. chabaudi AS MSP1 produced inconsistent effects, often parasitaemias were indistinguishable from controls despite significant anti-MSP1 antibody responses. The not unlikely contamination of MSP1 native preparations with erythrocyte (E) components was considered. Immunization with a mixture of the MSP1 C-terminus recombinant polypeptide and a Triton X-100 solubilized lysate of normal E resulted in enhanced control of parasitaemia, however, no effect was seen after administration of either component on its own. Co-immunization of E with the N-terminus polypeptide reversed the inhibition seen, on this occasion with this construct alone. PMID- 9223172 TI - Revisiting host/parasite interactions: molecular analysis of parasites collected during longitudinal and cross-sectional surveys in humans. AB - This paper summarizes the first conclusions arising from an analysis of parasite diversity in blood samples collected during longitudinal surveys conducted in Senegal. Parasite typing was carried out using a PCR-based molecular analysis of allelic polymorphism. The parasite populations circulating in the village of Dielmo during periods of intense transmission (when the inoculation rate was 0.5 1 infective bite/night) are characterized by a considerable allelic diversity of the MSP-1, MSP-2 and TRAP loci. A large proportion of blood samples contained several MSP-1 or MSP-2 alleles. In asymptomatic carriers, the complexity of the infections (number of alleles and genetic diversity of these alleles) was age dependent. In children, the trend was for a reduced complexity during clinical episodes. Molecular typing showed that successive clinical episodes experienced by children were caused by genetically distinct parasites. Longitudinal analysis of asymptomatic carriers indicated that in the absence of transmission, the same parasite types were carried for long periods, while rapidly changing profiles were observed during intense transmission season. The consequences of these findings on our understanding of acquired anti-parasite immunity in humans living in endemic are discussed. PMID- 9223173 TI - Age-related antibody profiles in Schistosoma haematobium infections in a rural community in Zimbabwe. AB - Antibody responses to soluble Schistosoma haematobium egg (SEA) and worm (SWA) antigens in a rural Zimbabwean study population were examined by ELISA. One hundred and sixteen S. haematobium infected and 124 non-infected individuals representing individuals greater than five years old, were included. Non-endemic control sera were obtained from a schistosomiasis non-endemic part of Zimbabwe and from Norwegian blood donors. A possible association between IgE antibody responses and resistance to S. haematobium infection was indicated by a negative correlation between IgE anti-SEA levels and intensity of S. haematobium infection, and by a positive correlation between IgE responses to SEA and SWA and age. Similarly, an association between IgA and anti-SWA and resistance to S. haematobium was suggested by a negative correlation to intensity of infection and a positive correlation with age. A probably association between IgM and IgG4 with susceptibility to S. haematobium infection was described; intensity of S. haematobium infection correlated positively with IgG, IgG4 and IgM responses to SEA and with IgG4 and IgM responses to SWA, also age correlated negatively with IgG4 and IgM responses to SEA and with IgG4 responses to SWA. These findings support the concept of IgG4 and IgM as blocking antibodies. Significant positive correlations between antibody responses to SEA and SWA suggests cross-reactivity between eggs and adult worms. In addition, the recognition by IgE and IgG4 of the same schistosomulum antigens in immunoblotting suggests competitions for the same antigens. PMID- 9223174 TI - Recombinant mouse IL-6 boosts specific serum anti-plasmodial IgG subtype titres and suppresses parasitaemia in Plasmodium chabaudi chabaudi infection. AB - C57BL/6 mice which were treated with recombinant mouse interleukin-6 (rmIL-6), 75 ng in 0.2 ml saline, i.p., 24 h before inoculation with Plasmodium chabaudi chabaudi and on days one, two and three could not control the primary acute parasitaemia but were able to suppress secondary parasitaemia significantly, peak of 3% and to reduce parasitaemia to subpatent levels within three weeks. Control C57BL/6 mice, which received saline injections only, developed two identical peaks of parasitaemia of at least 21% each, typical of a primary P. chabaudi chabaudi infection and cleared parasitaemia by day 28 post-inoculation. Serum levels of IL-6 measured in the first week by enzyme-linked immunosorbent assay were significantly higher (two-fold) in recipients compared to control mice. Anti plasmodial IgG1, 2a and 2b titres were four to 16 times higher in rmIL-6 recipients than in the control mice. Reduction of parasitaemia to subpatency during the secondary phase of P. chabaudi chabaudi infection in C57BL/6 mice is primarily antibody-mediated and rmIL-6 accelerates this process by boosting the levels of the required specific anti-plasmodial IgG subclasses of antibodies. PMID- 9223175 TI - Immune responses associated with protection in sheep vaccinated with a recombinant antigen from Taenia ovis. AB - This paper describes the evaluation of the protective antibody response of sheep to vaccination against Taenia ovis infection with a defined recombinant antigen (45W). Sera from 181 vaccinated sheep, collected prior to experimental challenge with T.ovis, were assessed for 45W specific IgA, IgG, IgG1, IgG2 and IgM levels and these results correlated with protection data. There were significant relationships (P < 0.001) between IgG, IgG1 and IgG2 titres and protection. Serum IgA levels did not correlate with protection and there were no significant levels of 45W specific IgM detected. Killing of several other taeniid cestodes has been shown to be complement mediated and the findings in this study are consistent with the involvement of this immune mechanism in 45W vaccinated sheep. A comparison of the adjuvants used in this study (saponin and oil in water) demonstrated that whereas both adjuvants stimulated the production of similar levels of 45W specific IgG1, the IgG2 response was significantly higher in sheep vaccinated with oil adjuvant. PMID- 9223176 TI - The role of anti-variable surface glycoprotein antibody responses in bovine trypanotolerance. AB - It has been reported that some breeds of cattle such as the N'Dama mount a more effective antibody response to the variable surface glycoprotein coat of trypanosomes and that this may contribute to their ability to control the infection. Thus we have investigated antibody responses to surface exposed epitopes of the variable surface glycoprotein in Trypanosoma congolense-infected N'Dama (trypanotolerant) and Boran (susceptible) cattle. Similar titres and isotypes were found in both N'Damas and Borans indicating that trypanotolerance is not associated with superior antibody-mediated destruction of trypanosomes. However, significant differences in antibody responses to cryptic VSG epitopes and non-trypanosome antigens were identified. Trypanosusceptible Boran cattle had low IgG1 responses to cryptic epitopes but high IgM responses to non-trypanosome antigens such as beta-galactosidase. In contrast the N'Dama cattle had significantly higher IgG1 responses to cryptic VSG epitopes and negligible responses to beta-galactosidase. These results indicate differences in the induction of anti-trypanosome immune responses between trypanotolerant and susceptible cattle infected with T. congolense. PMID- 9223177 TI - A multicopy modeling of the water distribution in macromolecular crystals. AB - A multicopy protocol is proposed for modeling macromolecular hydration using diffraction experimental data (X-ray or neutron) to search for a better description of delocalized water sites than that given by point water models. The model consists of one macro-molecule and several copies of each water molecule, refined simultaneously against diffraction data using molecular dynamics techniques. The protocol was applied to BPTI and an RNA tetradecamer. The sites defined by the different copies range from very ordered ones to continuous channels; they fit the density maps and agree with the diffraction amplitudes with an accuracy comparable with usual crystallographic methods. The delocalization of water in channels agrees with the high mobility observed in NMR experiments. PMID- 9223178 TI - Mean field analysis of FKBP12 complexes with FK506 and rapamycin: implications for a role of crystallographic water molecules in molecular recognition and specificity. AB - Mean field analysis of FKBP12 complexes with FK506 and rapamycin has been performed by using structures obtained from molecular docking simulations on a simple, yet robust molecular recognition energy landscape. When crystallographic water molecules are included in the simulations as an extension of the FKBP12 protein surface, there is an appreciable stability gap between the energy of the native FKBP12-FK506 complex and energies of conformations with the "native-like" binding mode. By contrast, the energy spectrum of the FKBP12-rapamycin complex is dense regardless of the presence of the water molecules. The stability gap in the FKBP12-FK506 system is determined by two critical water molecules from the effector region that participate in a network of specific hydrogen bond interactions. This interaction pattern protects the integrity and precision of the composite ligand-protein effector surface in the binary FKBP12-FK506 complex and is preserved in the crystal structure of the FKBP12-FK506-calcineurin ternary complex. These features of the binding energy landscapes provide useful insights into specific and nonspecific aspects of FK506 and rapamycin recognition. PMID- 9223179 TI - Measurement of water-amide proton exchange rates in the denatured state of staphylococcal nuclease by a magnetization transfer technique. AB - The rates of hydrogen exchange were measured in a "physiological" denatured state of staphylococcal nuclease using a NMR magnetization transfer experiment suitable for the measurement of exchange rates faster than 0.5 s-1. The results are compared with predicted exchange rates (kex) for the random coil state (Bai et al., Proteins 17:75-86, 1993). No protection factors (= predicted rate/measured rate) larger than 2.4 were observed, consistent with other NMR data which strongly suggest only small amounts of residual secondary structure in this denatured state. Systematically low protection factors (0.51 +/- 0.23) were found for Asp and Glu residues, while high protection factors were observed for Gly (1.60 +/- 0.60). We conclude that the predicted exchange rates (kex) may have an uncertainty of 2- to 3-fold. Thus, for denatured proteins only protection factors with a value of 5 or larger can be assigned structural significance. These results also demonstrate that multidimensional magnetization transfer NMR techniques are powerful tools in this research field due to its ability to measure rapidly exchanging protons (> 05 s-1) with high accuracy. PMID- 9223180 TI - Hydrophobic patches on protein subunit interfaces: characteristics and prediction. AB - Hydrophobic patches, defined as clusters of neighboring apolar atoms deemed accessible on a given protein surface, have been investigated on protein subunit interfaces. The data were taken from known tertiary structures of multimeric protein complexes. Amino acid composition and preference, patch size distribution, and patch contact complementarity across associating subunits were examined and compared with hydrophobic patches found on the solvent-accessible surface of the multimeric complexes. The largest or second largest patch on the accessible surface of the entire subunit was involved in multimeric interfaces in 90% of the cases. These results should prove useful for subunit design and engineering as well as for prediction of subunit interface regions. PMID- 9223181 TI - Predicting helical segments in proteins by a helix-coil transition theory with parameters derived from a structural database of proteins. AB - A novel helix-coil transition theory has been developed. This new theory contains more types of interactions than similar theories developed earlier. The parameters of the models were obtained from a database of 351 nonhomologous proteins. No manual adjustment of the parameters was performed. The interaction parameters obtained in this manner were found to be physically meaningful, consistent with current understanding of helix stabilizing/destabilizing interactions. Novel insights into helix stabilizing/destabilizing interactions have also emerged from this analysis. The theory developed here worked well in sorting out helical residues from amino acid sequences. If the theory was forced to make prediction on every residue of a given amino acid sequence, its performance was the best among ten other secondary structural prediction algorithms in distinguishing helical residues from nonhelical ones. The theory worked even better if one only required it to make prediction on residues that were "predictable" (identifiable by the theory); > 90% predictive reliability could be achieved. The helical residues or segments identified by the helix-coil transition theory can be used as secondary structural contraints to speed up the prediction of the three-dimensional structure of a protein by reducing the dimension of a computational protein folding problem. Possible further improvements of this helix-coil transition theory are also discussed. PMID- 9223182 TI - Refined solution structure of the anti-mammal and anti-insect LqqIII scorpion toxin: comparison with other scorpion toxins. AB - The solution structure of the anti-mammal and anti-insect LqqIII toxin from the scorpion Leiurus quinquestriatus quinquestriatus was refined and compared with other long-chain scorpion toxins. This structure, determined by 1H-NMR and molecular modeling, involves an alpha-helix (18-29) linked to a three-stranded beta-sheet (2-6, 33-39, and 43-51) by two disulfide bridges. The average RMSD between the 15 best structures and the mean structure is 0.71 A for C alpha atoms. Comparison between LqqIII, the potent anti-mammal AaHII, and the weakly active variant-3 toxins revealed that the LqqIII three-dimensional structure is closer to that of AaHII than to the variant-3 structure. Moreover, striking analogies were observed between the electrostatic and hydrophobic potentials of LqqIII and AaHII. Several residues are well conserved in long-chain scorpion toxin sequences and seem to be important in protein structure stability and function. Some of them are involved in the CS alpha beta (Cysteine Stabilized alpha-helix beta-sheet) motif. A comparison between the sequences of the RII rat brain and the Drosophila extracellular loops forming scorpion toxin binding-sites of Na+ channels displays differences in the subsites interacting with anti-mammal or anti-insect toxins. This suggests that hydrophobic as well as electrostatic interactions are essential for the binding and specificity of long-chain scorpion toxins. PMID- 9223183 TI - A potential processing enzyme in prokaryotes: oligopeptidase B, a new type of serine peptidase. AB - Basic amino acid pairs in polypeptides represent important markers for processing enzymes to produce biologically active products. Such enzymes related to the serine peptidase subtilisin have recently been identified in eukaryotes. Herein is described and kinetically characterized a new type of processing enzyme, oligopeptidase B, which is encountered in the prokaryote Escherichia coli, and belongs to the prolyl oligopeptidase family of serine peptidase. The enzyme hydrolyzes the peptides at the carboxy end of dibasic sites by two orders of magnitude faster with respect to monobasic substrates. The kcat/K(m) is extremely high, 63 microM-1 s-1, for the substrate benzyloxycarbonyl-L-arginyl-L-arginyl-7 (4-methylcoumaryl)amide. The bell-shaped pH dependence of the rate constant is perturbed by some ionizing group(s). This effect is abolished at 1 M NaCl. In addition, high ionic strength inhibits the reaction considerably by increasing K(m), which is indicative of an electrostatic interaction between the arginyl residues and the enzymatic carboxy groups. In distinction from that found with most serine endopeptidases, kinetic deuterium isotope measurements with oligopeptidase B indicate that the rate-limiting step of the reaction is a physical step rather than a chemical one characterized by general acid/base catalysis. The present result will contribute to our understanding of the processing phenomena in prokaryotes, as well as in higher organisms. PMID- 9223184 TI - Picosecond dynamical changes on denaturation of yeast phosphoglycerate kinase revealed by quasielastic neutron scattering. AB - Quasielastic neutron scattering experiments performed on yeast phosphoglycerate kinase in the native form and denatured in 1.5 M guanidinium chloride reveal a change in the fast (picosecond time scale) diffusive internal dynamics of the protein. The momentum and energy transfer dependences of the scattering for both states are fitted by an analytical model in which, on the experimentally accessible picosecond time scale and angstrom length scale, the dynamics of a fraction of the nonexchangeable hydrogens in the protein is described as a superposition of vibrations with uniform diffusion in a sphere, the rest of the hydrogens undergoing only vibrational motion. The fraction diffusing changes, from approximately 60% in the native protein to approximately 82% in the denatured protein. The radius of the sphere also changes slightly, from approximately 1.8 A in the native protein to approximately 2.2 A in the denatured protein. Possible implications of these results for the general protein folding problem are discussed. PMID- 9223185 TI - Automated multiple analysis of protein structures: application to homology modeling of cytochromes P450. AB - A computational strategy for homology modeling, using several protein structures comparison, is described. This strategy implies a formalized definition of structural blocks common to several protein structures, a new program to compare these structures simultaneously, and the use of consensus matrices to improve sequence alignment between the structurally known and target proteins. Applying this method to cytochromes P450 led to the definition of 15 substructures common to P450cam, P450BM3, and P450terp, and to proposing a 3D model of P450eryF. PMID- 9223186 TI - Pfam: a comprehensive database of protein domain families based on seed alignments. AB - Databases of multiple sequence alignments are a valuable aid to protein sequence classification and analysis. One of the main challenges when constructing such a database is to simultaneously satisfy the conflicting demands of completeness on the one hand and quality of alignment and domain definitions on the other. The latter properties are best dealt with by manual approaches, whereas completeness in practice is only amenable to automatic methods. Herein we present a database based on hidden Markov model profiles (HMMs), which combines high quality and completeness. Our database, Pfam, consists of parts A and B. Pfam-A is curated and contains well-characterized protein domain families with high quality alignments, which are maintained by using manually checked seed alignments and HMMs to find and align all members. Pfam-B contains sequence families that were generated automatically by applying the Domainer algorithm to cluster and align the remaining protein sequences after removal of Pfam-A domains. By using Pfam, a large number of previously unannotated proteins from the Caenorhabditis elegans genome project were classified. We have also identified many novel family memberships in known proteins, including new kazal, Fibronectin type III, and response regulator receiver domains. Pfam-A families have permanent accession numbers and form a library of HMMs available for searching and automatic annotation of new protein sequences. PMID- 9223187 TI - Structural consensus in ligand-protein docking identifies recognition peptide motifs that bind streptavidin. AB - Computational structure prediction of streptavidin-peptide complexes for known recognition sequences and a number of random di-, tri-, and tetrapeptides has been conducted, and mechanisms of peptide recognition with streptavidin have been investigated by a new computational protocol. The structural consensus criterion, which is computed from multiple docking simulations and measures the accessibility of the dominant binding mode, identifies recognition motifs from a set of random peptide sequences, whereas energetic analysis is less discriminatory. The predicted conformations of recognition tripeptide and tetrapeptide sequences are also in structural harmony and composed of peptide fragments that are individually unfrustrated in their bound conformation, resulting in a minimally frustrated energy landscape for recognition peptides. PMID- 9223188 TI - Calculation of pathways for the conformational transition between the GTP- and GDP-bound states of the Ha-ras-p21 protein: calculations with explicit solvent simulations and comparison with calculations in vacuum. AB - The transitions between the water-equilibrated structures of the GTP and GDP forms of Ha-ras-p21 have been calculated by using the targeted molecular dynamics (TMD) method (Schlitter et al., Mol. Sim. 10:291-309, 1993) both in vacuo and with explicit solvent simulation. These constrained molecular dynamics calculations result in different pathways, depending on the nucleotide bound. Each pathway consists in a sequence of transitions affecting six segments of the protein, four of them forming a hydrophilic cleft around the nucleotide. The transitions are initiated by the removal or introduction of the gamma-phosphate of the nucleotide and proceed sequentially, crossing several low-energy transition states. The movements are transmitted either by direct interactions between the segments or through the nucleotide. The GTP to GDP pathway is initiated by the removal of the nucleotide gamma-phosphate. This gives some space to Gly12, Gly13, and Val14. Their movement is transmitted to the target recognition domain and the switch II region, forcing these segments to adopt another position. In a second step the target recognition domain and the switch II region undergo conformational transitions to reach an intermediate conformation. Finally, there is a relaxation of the target recognition domain to its final state that forces the switch II region to reach its target conformation. The calculated pathways allow the identification of many residues that play an important role in the conformational changes, explain the altered transformation properties of some, and suggest mutations to alter the pathway. PMID- 9223189 TI - Crystallization and preliminary crystallographic analysis of the N-terminal actin binding domain of human fimbrin. AB - We have crystallized the N-terminal actin binding domain (ABD1) of human fimbrin, a representative member of the largest class of actin crosslinking proteins. Diffraction from these crystals is consistent with the orthorhombic space group P2(1)2(1)2(1) (a = 50.03 A, b = 61.24 A, c = 102.30 A). These crystals contain one molecule in the asymmetric unit and diffract to at least 1.9 A resolution. The crystal structure of ABD1 will be the first structure of an actin crosslinking domain. PMID- 9223190 TI - Crystallization and preliminary X-ray diffraction analysis of bovine seminal plasma PDC-109, a protein composed of two fibronectin type II domains. AB - PDC-109 is a 13 kDa glycoprotein and the major phosphorylcholine- and heparin binding protein of bull seminal plasma. It is built by an acidic 23-residue N terminal sequence followed by a tandem of fibronectin type II domains. Full length PDC-109 was crystallized in complex with o-phosphorylcholine by vapor diffusion in sitting drops. Crystals grew to maximal size of 0.5 x 0.3 x 0.2 mm3, diffract x-rays beyond 2.6 A resolution, and belong to space group P321 with unit cell dimensions a = b = 93.6 A, c = 52.7 A. PMID- 9223191 TI - Crystallization and preliminary cryogenic X-ray diffraction analyses of protein L isoaspartyl O-methyltransferase from human fetal brain. AB - Recombinant human fetal brain protein L-isoaspartyl O-methyltransferase, EC 2.1.1.77, was crystallized in PEG 8000 with adenosine homocysteine by a macroseeding technique. The space group was P2(1) with a = 47.4 A, b = 53.9 A, c = 48.7 A and beta = 116.4 degrees for cryofrozen crystals at 90 K. The crystals diffracted to 2.1 A and have one molecule per asymmetric unit. PMID- 9223192 TI - Back pain following epidural anesthesia with 2-chloroprocaine. PMID- 9223193 TI - Back pain following epidural anesthesia with 2-chloroprocaine (EDTA-free) or lidocaine. AB - BACKGROUND AND OBJECTIVES: Severe lumbar pain following epidural injection of 2 chloroprocaine is usually associated with the Nesacaine-MPF solution available in the United States. The purpose of this study was to determine if the solution distributed in Canada (Nesacaine-CE), which contains calcium disodium edetate (0.1 mg/mL) and sodium bisulfite (0.7 mg/mL) but no disodium ethylenediaminetetraacetic acid, is associated with back pain or spasm when compared with epidural lidocaine. METHODS: With use of a prospective, double blind, randomized design, 30 patients scheduled to undergo outpatient knee arthroscopy under epidural anesthesia were divided into two groups to received 30 mL of either Nesacaine-CE 3% (group A) or lidocaine 1.33% (group B). Postoperative pain in the lumbar area was assessed twice by a 10-cm visual analog scale (VAS) before patients left the hospital and 24 hours later by phone. The lumbar area was palpated to search for muscle spasm before discharge from hospital. RESULTS: More patients receiving Nesacaine-CE than receiving lidocaine suffered from back pain in the recovery room (four vs none P = .03) and before leaving the hospital (nine vs one P = .001). Higher VAS scores (mean +/- SE) were obtained after Nesacaine CE then after lidocaine in the recovery room (0.5 +/- 0.24 vs 0.0 +/- 0.0, p = .049) and before leaving the hospital (1.8 +/- 0.5 vs 0.1 +/- 0.1, P = .001). No difference existed 24 hours later between the two groups with regard to the prevalence of back pain or VAS scores. No muscle spasm was detected. CONCLUSION: No cases of severe backache were observed. However, epidural Nesacaine-CE 3% was associated with mild back pain, generally confined to the area of needle insertion, when compared with lidocaine 1.33%. PMID- 9223194 TI - Distribution of local anesthetic solution in retromediastinal block. Preliminary experimental results. AB - BACKGROUND AND OBJECTIVES: Interpleural anesthesia blocks pain perception from the thoracoabdominal wall without impairment of leg function. Bilateral interpleural anesthesia is not recommended because of possible bilateral impairment of respiratory function. Infiltration of the retromediastinum with local anesthetic might cause bilateral thoracoabdominal somatic block and block of sympathetic afferents from the abdominal cavity without impairing respiration. METHOD: Distribution of stained fluid was studied after injection into the retromediastinum through a catheter placed about 10 cm cephalad to the diaphragm via the esophageal hiatus in three human cadavers of normal size and in six anesthetized pigs of 20-30 kg. In the pigs serum levels of bupivacaine were measured after injection of 10 mL of 0.5% bupivacaine stained with 1 mL of methylene blue. RESULTS: The injected Dye stained intercostal nerves 6-11 in cadavers and 5-12 in pigs symmetrically on both sides, along with the adjacent parts of the sympathetic chain and both vagal nerves but not the phrenic nerves. During the sampling period of 50-60 minutes, bupivacaine serum concentrations rose slowly to a maximum of 4.2 micrograms/mL. CONCLUSIONS: Block of pain perception from the abdominal wall and cavity is possible by injection of local anesthetic into the retromediastinum via a catheter introduced through the esophageal diaphragm hiatus. The block would not be expected to impair respiratory or leg function. Its efficacy and safety have yet to be established. PMID- 9223195 TI - Sensory and sympathetic block during interpleural analgesia. AB - BACKGROUND AND OBJECTIVES: Interpleural analgesia is an effective method for pain relief after upper abdominal surgery. To examine whether the analgesic effect is obtained by block of the intercostal nerves, we assessed the analgesic efficacy of the block, the skin sensitivity, and indices of sympathetic outflow over the trunk. METHODS: Interpleural analgesia was instituted at the end of open cholecystectomy in 20 patients 24-81 years of age (mean, 42 years). After a washout period of 8 hours, the analgesic effect was tested 5-12 times during the postoperative follow-up period by using a visual analogue scale before and 20 minutes after injection of 20 mL of bupivacaine 0.25%. Temperature and pain sensations were tested on the day after surgery, and in nine of the patients, the cutaneous blood flow over the trunk was studied by an electronic thermometer, laser Doppler flowmetry, and fluorescein flowmetry. In addition, the conduction velocity in the phrenic nerve was studied in four of the patients. RESULTS: Interpleural analgesia significantly reduced the median visual analogue score from of 5.7 (range 2-10) to 1.1 (range, 0-4). Although the analgesic effect was very good in all patients, half of them still showed skin sensitivity to pain and temperature. Cutaneous blood flow did not change, which showed than block of the intercostal nerves was incomplete. The phrenic nerve was not affected. CONCLUSION: The incomplete cutaneous sensory and sympathetic block indicates that the analgesic effect of interpleural analgesia cannot be explained by retrograde diffusion of the local anesthetic solution into the intercostal nerves alone. PMID- 9223196 TI - Effect of thoracic epidural anesthesia on spontaneous postinfarction ventricular dysrhythmia in awake dogs. AB - BACKGROUND AND OBJECTIVES: Sympathetic neural activity contributes to the genesis of ventricular ectopic activity, particularly in the setting of myocardial ischemia and infarction, so thoracic epidural anesthesia should diminish ventricular ectopy by blocking sympathetic innervation of the heart. However, the possible antidysrhythmic effect of epidural anesthesia has been studied only in the presence of general anesthesia. We therefore examined changes in spontaneous postinfarction ventricular dysrhythmia during thoracic epidural anesthesia in awake dogs. METHODS: A survivable myocardial infarction was created by two-stage ligation of the left anterior descending coronary artery. The following day, multifocal idioventricular tachycardia was the predominant cardiac rhythm. Lidocaine was administered either by thoracic epidural catheter to achieve block of at least the first five thoracic segments or intravenously as a control for direct effects, without concurrent general anesthesia or sedation. Electrocardiographic recordings were analyzed for the number of ventricular ectopic and sinoatrial depolarizations. RESULTS: Epidural and intravenous administration both produced plasma lidocaine concentrations of about 2 mg/mL. There was no change in rhythm following intravenous lidocaine. During epidural anesthesia, total ectopic beats per minute decreased from 167 +/- 8 to 135 +/- 14 (mean +/- SE, P < .05), and the dysrhythmic ratio (ventricular beats/total beats) decreased from 0.93 +/- 0.03 to 0.81 +/- 0.08 (P < .05). However, ventricular tachydysrhythmia remained the predominant rhythm. CONCLUSIONS: Epidural block modestly reduces spontaneous ventricular dysrhythmia in a perioperative setting in dogs following a large myocardial infarction. These findings do not support the choice of thoracic epidural anesthesia for the purpose of preventing or decreasing severe ventricular dysrhythmia. PMID- 9223197 TI - Sympathetic block during spinal anesthesia in volunteers using lidocaine, tetracaine, and bupivacaine. AB - BACKGROUND AND OBJECTIVES: Spinal anesthesia to high thoracic dermatomes is alleged to result in almost complete block of all sympathetic efferent nerves. To examine the degree of sympathectomy during spinal anesthesia, the sympathetic response to a cold pressor test (CPT) applied to unblocked dermatomes before and during spinal anesthesia was measured with use of three different local anesthetics. METHODS: Twelve healthy volunteers were studied in a randomized and double-blind fashion on three separate occasions. In random order, each volunteer received approximately equipotent spinal doses of tetracaine 15 mg, bupivacaine 15 mg, and lidocaine 100 mg in hyperbaric solutions. Prior to and 30 minutes after spinal injection of local anesthetic, a CPT was applied for 2 minutes, and changes from baseline resting conditions in five physiologic variables were measured. RESULTS: The CPT 1 given before anesthetic administration resulted in an increase in heart rate, mean arterial pressure, cardiac index, and plasma concentrations of norepinephrine and epinephrine. Spinal anesthesia to a median level of T3 resulted in a decrease in mean arterial pressure by 10-12% but did not significantly decrease the other variables. Spinal anesthesia did not change the increase in heart rate or cardiac index in response to the second CPT, but the increase in mean arterial pressure was attenuated compared to the CPT before anesthesia. No increase in norepinephrine or epinephrine concentration was observed during the CPT given during spinal anesthesia. There was no significant relationship between level of analgesia and sympathetic response to stress. CONCLUSIONS: Spinal anesthesia with hyperbaric solutions of tetracaine 15 mg, bupivacaine 15 mg, and lidocaine 100 mg attenuated sympathetic function but did not produce complete sympathectomy. The effects were independent of the local anesthetic used. PMID- 9223198 TI - Patterns of pain induced by distending the thoracic zygapophyseal joints. AB - BACKGROUND AND OBJECTIVES: The study was designed to investigate the patterns of referral pain associated with the thoracic zygapophyseal joints (C7-T1 to T2-3, T11-12). METHODS: In 15 patients who had back pain suspected to be of zygapophyseal origin and for whom pain was relieved by injection of local anesthetic into the joints under fluoroscopic guidance, the zygapophyseal joints from C7-T1 to T2-3, T11-12 were distended by injecting contrast medium. If injection reproduced typical pain in the patient, the patient was asked to describe the distribution of pain, which was recorded on a body diagram. RESULTS: A total of 21 joints were studied. Composite pain distribution maps for the thoracic zygapophyseal joints from C7-T1 to T2-3 and for the T11-12 joint were drawn. The distribution of referred pain was as follows: pain in the suprascapular region was referred from C7-T1 and Th1-2; pain in the superior angle of the scapula from C7-T1 and T1-2; pain in the mid-scapular region from C7 T1, T1-2, and Th2-3; and pain in the paravertebral region around the site of injection and the area over the iliac crest from T11-12. CONCLUSIONS: The referred pain distribution for joints C7-T1 to Th2-3 showed significant overlap. PMID- 9223199 TI - Postoperative analgesia and antiemetic efficacy after subarachnoid neostigmine in orthopedic surgery. AB - BACKGROUND AND OBJECTIVES: The efficacy of operatively administered spinal neostigmine to provide analgesia and that of different antiemetics to prevent neostigmine-related nausea and vomiting were evaluated in patients undergoing tibial or ankle reconstruction. METHODS: One hundred patients were randomized to five groups (n = 20). The intravenous antiemetic test drug (except propofol) was given as premedication in the holding room, after intravenous midazolam, 0.05 mg/kg. The subarachnoid drugs administered were 20 mg bupivacaine (0.5%) in conjunction with 100 micrograms neostigmine, except for the saline group (S group), which received bupivacaine and saline. The S group, the neostigmine group (N group), and the propofol group (P group) received saline as the intravenous test drug. The droperidol group (D group) received intravenous droperidol 0.5 mg, and the metoclopramide group (M group) received intravenous metoclopramide 10 mg. The P group had a continuous intravenous propofol infusion (2-4 mg/kg/hr), started 10 minutes after the spinal injection. Nausea, emetic episodes, and the need for analgesic (disclofenac) or antiemetic medication were recorded for the first 24 hours following surgery and scored by a 10-cm visual analog scale (VAS). RESULTS: Subarachnoid neostigmine 100 micrograms did not affect subarachnoid bupivacaine analgesia as measured by time to first rescue analgesic in most patients, but it decreased the overall 24-hour visual analog scale (VAS) scores and the need for postoperative analgesics in 24 hours (P < .001). The incidence of intraoperative nausea and vomiting was higher in the N, D, and M groups than in the S group (P < .001). Following surgery, the 3-hour VAS assessment for emesis was higher for the N, P, and M groups than for the S group (P < .05). The overall 24-hour assessment was similar among groups. CONCLUSIONS: Subarachnoid neostigmine reduced postoperative pain scores and analgesic requirements. Whether it prolonged the duration of action of diclofenac or enhanced the mechanisms involved in spinal analgesia cannot be determined from these data. Although propofol and droperidol appeared to be more effective during and after surgery, respectively, all neostigmine groups were associated with a high consumption of antiemetics. PMID- 9223200 TI - Ketorolac as an adjunct to patient-controlled morphine in postoperative spine surgery patients. AB - BACKGROUND AND OBJECTIVES: This randomized double-blind study was designed to determine whether administration of ketorolac either on schedule or as a component of patient-controlled analgesia (PCA) to patients who have undergone spinal stabilization would decrease PCA morphine use, decrease side effects, and/or enhance analgesia. METHODS: Eighty inpatients undergoing spine stabilization by one surgeon were evaluated after excluding patients with contraindications to the use of ketorolac or morphine. All patients received PCA morphine. The patients were divided into four groups, which were given either intravenous saline (control group); intravenous ketorolac 15 mg every 6 hours; intravenous ketorolac 30 mg every 6 hours; or ketorolac added to the PCA morphine on a milligram per milligram basis. The outcome measures included pain scores. 24 hour morphine use, level of sedation, and side effect profile at six times during the first 24 postoperative hours. RESULTS: The total dose of morphine (P < .0001) and the cumulative doses for each of the six time periods (P varied between .02 and .0001 for the six time periods) were significantly higher in the control group than in the other three groups. There were no differences in doses administered by the other three groups. The pain scores were also significantly higher in the control group than in the other three groups, with no differences in pain scores among the other three. The sedation scores were higher (ie, patients more sedated) in the control group than in the other three groups at two of the six time periods (periods I and 6; P < .001). CONCLUSIONS: Ketorolac should be as a component of the PCA morphine in patients undergoing spine stabilization surgery. This results in decreased morphine consumption, decreased somnolence, and enhanced analgesia in comparison with patients who do not receive ketorolac. PMID- 9223201 TI - Addition of morphine to intra-articular bupivacaine does not improve analgesia following knee joint replacement. AB - BACKGROUND AND OBJECTIVES: In an effort to further decrease postoperative opioid requirements and improve analgesia in patients undergoing elective knee joint replacement, a study was made of the effectiveness of adding morphine to an intra articular bupivacaine injection given immediately following surgery. METHODS: In random, double-blind fashion, 75 patients received a 31-mL intra-articular injection consisting of either 30 mL 0.5% bupivacaine with 1:200,000 epinephrine and 1 mL normal saline (group BUP), 30 mL 0.5% bupivacaine with 1:200,000 epinephrine and 1 mg (1 mL) preservative-free morphine (group BUP-MORPH), or 30 mL normal saline with 1:200,000 epinephrine and 1 mg preservative-free morphine (group MORPH). Postoperative analgesia was supplied by patient controlled analgesia (PCA) with morphine. Patients were assessed at 1, 2, 4, and 24 hours for pain (visual analog and verbal rating scales), morphine utilization, and side effects. Knee range of motion was measured before operation and at hospital discharge. RESULTS: There was no difference among the three groups in PCA morphine requirements, pain scores by either scale, range of motion, or incidence of side effects, including somnolence, urinary retention, nausea and vomiting, and pruritus. CONCLUSION: The addition of 1 mg morphine to an intra-articular injection of 30 mL 0.5% bupivacaine with 1:200,000 epinephrine given at wound closure does not improve analgesia in patients undergoing elective knee joint replacement. PMID- 9223202 TI - Local anesthetic infusion through nerve sheath catheters for analgesia following upper extremity amputation. Clinical report. AB - BACKGROUND AND OBJECTIVES: Reports about the efficacy of local anesthetic perfusion of nerve stumps following lower extremity amputation are conflicting. We report our experience with this technique following amputation of the upper extremity. METHODS: Six consecutive patients undergoing proximal upper extremity amputations (four forequarter amputations and two shoulder disarticulations) for malignancy were prospectively observed. In all patients, catheters were placed within the amputated nerve sheaths at the conclusion of the procedure. Bupivacaine. 0.25%, was administered through each catheter as a bolus and then as a continuous infusion for at least 72 hours after surgery. Narcotic usage, level of pain as reported verbally, and presence of phantom limb pain during the infusion were recorded. For at least 1 year after operation, data were gathered on the presence of phantom limb pain and its intensity during each follow-up visit. RESULTS: Complete analgesia was achieved in all patients by postoperative day 2. Narcotic usage was low. Three of the six patients reported phantom limb pain during follow-up evaluation. CONCLUSIONS: Continuous local anesthetic perfusion of amputated nerves via a catheter placed under direct vision provided excellent postoperative analgesia. The incidence of phantom limb pain for cancer patients did not differ from that previously reported but was easily managed pharmacologically. The technique may also be efficacious for traumatic amputations. PMID- 9223203 TI - Continuous axillary brachial plexus block for postoperative pain relief. Intermittent bolus versus continuous infusion. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to compare the efficacy and safety of continuous axillary brachial plexus block by infusion and by intermittent injection of bupivacaine 0.25% in the management of postoperative pain. METHODS: Two methods of continuous axillary brachial plexus block for pain management were compared in a prospective randomized trial. Twenty 16- to 62-year old male patients (ASA I and II), undergoing microsurgery, received 0.25% bupivacaine following surgery, by either continuous infusion (group C, n = 10), or intermittent hourly bolus (group B, n = 10), via an axillary brachial plexus catheter inserted prior to operation. Efficacy assessments and analysis of plasma bupivacaine levels were performed for up to 38 hours postsurgery. RESULTS: There was no difference between groups with respect to pain scores, degree of motor block, or supplemental narcotic requirements. Plasma bupivacaine levels were higher in group C, with the difference reaching significance after 26 hours (means +/- SEM: C = 1.03 +/- 0.10 micrograms/mL, B = 0.73 +/- 0.08 microgram/mL, P < .05). There were no significant differences in cumulative bupivacaine doses or infusion rates between the groups. Complications included one case of axillary artery puncture at insertion, two unplanned premature catheter removals, and a self-limited grand mal convulsion in a known epileptic. CONCLUSIONS: Overall, both techniques provided safe and effective postoperative analgesia. As compared with continuous infusion, intermittent bolus administration resulted in lower plasma bupivacaine levels despite similar infusion rates. PMID- 9223204 TI - Intraneural lidocaine uptake compared with analgesic differences between pregnant and nonpregnant rats. AB - BACKGROUND AND OBJECTIVES: Pregnant patients need less local anesthetic in order to obtain the same quality of functional block as nonpregnant patients. Our goal was to demonstrate a similarly increased functional susceptibility to local anesthetics in the awake pregnant rat during peripheral nerve block and to investigate the pharmacokinetic and/or pharmacodynamic mechanisms responsible for this phenomenon. METHODS: Radiolabeled lidocaine uptake was determined in vivo during sciatic nerve block with 0.1 ml of 1% lidocaine in the nerves of nine pregnant and five nonpregnant female rats and six male rats at the return of deep pain sensation, assessed by withdrawal of the hindlimb from a brief squeeze of a digit with serrated forceps. During recovery from complete functional block, the time at which deep pain returned and the amount of lidocaine in the nerve at that time were compared among the three groups of rats. Lidocaine content was also determined in vitro after exposure of ensheathed sciatic nerves from pregnant and nonpregnant rats to a 0.2% lidocaine bath for specified times. RESULTS: Full block of function developed in all groups within 6 minutes of the lidocaine injection and lasted significantly longer in pregnant rats than in nonpregnant and male rats (49.0 +/- 3.3 vs 34.0 +/- 3.1 and 32.0 +/- 1.3 minutes mean +/- SEMI, respectively. At the time of deep pain return, the intraneural lidocaine content of pregnant rats was significantly lower than that of nonpregnant and male rats (2.2 +/- 0.25 vs 3.9 +/- 0.7 and 3.7 +/- 0.6 nmoles/mg of wet nerve, respectively). No difference in lidocaine uptake kinetics between P and NP nerves was observed in vitro. CONCLUSIONS: Block of peripheral neural function is prolonged in pregnant rats, and lidocaine content in the nerve is lower at a specific stage of neural block. These results are consistent with a pharmacodynamic mechanism for increased susceptibility to lidocaine neural block during pregnancy. PMID- 9223205 TI - Bilateral fascia iliaca catheters for postoperative pain control after bilateral total knee arthroplasty: a case report and description of a catheter technique. AB - BACKGROUND AND OBJECTIVES: The pain following total knee arthroplasty can be associated with significant morbidity, especially in the elderly. Regional anesthetic techniques attenuate or eliminate postoperative pain, which may reduce this morbidity. METHODS: A 74-year-old patient with history of an epidural abscess underwent elective bilateral total knee arthroplasty for degenerative joint disease. Bilateral lumbar plexus catheters were placed via the fascia iliaca compartments. Lidocaine was infused postoperative through both catheters, and serum lidocaine levels were followed. RESULTS: The patient received significant postoperative pain relief based on physical and subjective examination. There were no complications or untoward effects related to the technique. CONCLUSION: Lumbar plexus blockade with continuous local anesthetic infusion via the fascia iliaca compartment is an effective means of providing postoperative analgesia after total knee arthroplasty when epidural analgesia is contraindicated. PMID- 9223207 TI - Late recurrence of postdural puncture headache. AB - BACKGROUND AND OBJECTIVES: Postdural puncture headache can recur following an epidural blood patch but usually does so within the first week. However, a late recurrence was encountered in a 31-year-old woman with probable lupus vasculitis. METHODS: The patient underwent a diagnostic lumbar puncture and suffered a postdural puncture headache. She was treated with an epidural blood patch and had complete resolution of her headache. Five weeks later, she suffered a sudden recurrence of an identical headache and underwent a repeat epidural blood patch. RESULTS: The second headache, like the first, was immediately and completely resolved. CONCLUSIONS: A review of risk factors for postdural puncture headache and the role of an epidural blood patch suggests a possible explanation for the late recurrence in this case. PMID- 9223206 TI - Fetal bradycardia and uterine hyperactivity following subarachnoid administration of fentanyl during labor. AB - BACKGROUND AND OBJECTIVES: Changes in uterine tone have been postulated as the cause of fetal bradycardia following subarachnoid administration of fentanyl for labor analgesia. Such a case occurred in a 20-year-old parturient with an intrauterine pressure catheter in place. METHODS: The patient was given intravenous terbutaline, after which contractions ceased for 20-30 minutes and then resumed. RESULTS: The patient underwent successful cesarean delivery. Retrospective analysis of the data revealed a significant increase in uterine tone and contractions following fentanyl administration. CONCLUSIONS: This case supports the view that changes in uterine tone, producing a hyperdynamic contractile state and a resulting decrease in uteroplacental perfusion, may explain the fetal bradycardia following subarachnoid opioid administration. Cases that do not resolve spontaneously may respond to intravenous terbutaline. PMID- 9223208 TI - Comment on peroneal afferent nerve discharges and the formalin test. PMID- 9223209 TI - Neurolytic interventions for upper extremity spasticity associated with head injury. PMID- 9223210 TI - A different approach in demonstrating the mechanism of intravenous regional anesthesia. PMID- 9223211 TI - Does the interscalene block require paresthesia? PMID- 9223212 TI - "No paresthesias-no anesthesia," the nerve stimulator or neither? PMID- 9223213 TI - Lateral decubitus and the nonaxillary roll. PMID- 9223214 TI - Pharmacokinetics of methanol and formate in female cynomolgus monkeys exposed to methanol vapors. AB - The 1990 Clean Air Act Amendments contain mandates for reduced automotive emissions and add new requirements for the use of alternative fuels such as methanol to reduce certain automotive pollutants. Methanol is acutely toxic in humans at relatively low doses, and the potential for exposure to methanol will be increased if it is used in automotive fuel. Formate is the metabolite responsible for neurotoxic effects of acute methanol exposure. Since formate metabolism is dependent on folate, potentially sensitive folate-deficient subpopulations, such as pregnant women, may accumulate formate and be at higher risk from low-level methanol exposure. Our objective was to determine the pharmacokinetics of 14C-methanol and 14C-formate in normal and folate-deficient monkeys after exposure to 14C-methanol vapors at environmentally relevant concentrations: below the threshold limit value (TLV), at the TLV of 200 parts per million (ppm), and above the TLV. Four normal adult female cynomolgus monkeys were individually anesthetized with isoflurane, and each was exposed by endotracheal intubation to 10, 45, 200, or 900 ppm 14C-methanol for 2 hours. Concentrations of the inhaled and exhaled 14C-methanol, blood concentrations of 14C-methanol and 14C-formate, exhaled 14C-carbon dioxide (14CO2), and respiratory parameters were measured during exposure. After exposure, 14C-methanol and 14CO2 exhaled, 14C-methanol and 14C-formate excreted in urine, and 14C-methanol and 14C formate in blood were quantified. The amounts of exhaled 14C-methanol and 14CO2, blood concentrations of 14C-methanol and 14C-formate, and 14C-methanol and 14C formate excreted in urine were linearly related to methanol exposure concentration. For all exposures, blood concentrations of 14C-methanol-derived formate were 10 to 1000 times lower than endogenous blood formate concentrations (100 to 200 mM) reported for monkeys and were several orders of magnitude lower than levels of formate known to be toxic. Since the metabolism of formate in primates depends on the availability of tetrahydrofolate, the same four monkeys were next placed on a folate-deficient diet until folate concentrations in red blood cells consistent with moderate folate deficiency (29 to 107 ng/mL) were achieved. Monkeys were then reexposed to the highest exposure concentration, 900 ppm 14C-methanol, for a similar 2-hour period, and again the pharmacokinetic data described above were obtained. Even with a reduced folate status, monkeys exposed to 900 ppm methanol for 2 hours had peak concentrations of methanol-derived formate that were well below the endogenous levels of formate. Although these results represent only a single exposure and therefore preclude broad generalizations, they do suggest the body contains sufficient folate stores to effectively detoxify small doses of methanol-derived formate from exogenous sources, such as those that might occur during normal use of automotive fuel. PMID- 9223215 TI - The coming of age of the GDNF family and its receptors: gene delivery in a rat Parkinson model may have clinical implications. PMID- 9223216 TI - Alzheimer transgenic mouse models come of age. PMID- 9223217 TI - Monitoring secretion in real time: capacitance, amperometry and fluorescence compared. AB - Techniques for measuring exocytosis, endocytosis and vesicle cycling in living cells in real time have resulted in a rapid expansion in the knowledge of these processes in neurons and other secretory cells. Several experimental approaches, developed during the past decade, have played key roles in this expansion. In this review we focus on three techniques: electrophysiological methods for monitoring membrane capacitance, electrochemical methods for detecting released secretory contents and optical methods for imaging membranes of endosomes and recycled vesicles that are stained with fluorescent dyes. Each technique has contributed unique and complementary information about the vesicle cycle, advancing our knowledge of the kinetics of membrane fusion and retrieval, the identity of the secretory contents and the spatial patterns and directional pathways involved in secretory membrane recycling. Naturally, each technique has inherent limitations; some of these shortcomings have recently been resolved by using more than one method simultaneously. PMID- 9223218 TI - p75NTR and apoptosis: Trk-dependent and Trk-independent effects. AB - The ongoing dissection of the roles of p75NTR and TrkA, -B and -C in neurotrophin signaling has generated a number of apparent paradoxes. Limiting consideration to the role of p75NTR in cell death, a theory is proposed that is based on the following postulates: (1) that p75NTR displays a pro-apoptotic intrinsic (ligand independent, Trk-independent) receptor effect (IRE), which is inhibited by ligand binding; (2) that p75NTR and TrkA exhibit mutual repression of signaling; and (3) that p75NTR and TrkA are required for the efficient generation of high-affinity NGF binding sites. PMID- 9223219 TI - Variation and selection in neural function. PMID- 9223220 TI - Variation and selection in neural function. PMID- 9223221 TI - Y-receptor subtypes--how many more? AB - The Y-receptors belong to the G protein-coupled receptor superfamily and mediate a wide variety of physiological effects, such as regulation of blood pressure, anxiety, memory retention, hormone release and food intake. Since the first human Y-receptor was cloned in 1992, the search for additional subtypes has been an area of intense study. Recently four new NPY-receptor subtypes have been isolated, revealing surprisingly limited sequence identity with values as low as 30%. Several reports indicate further heterogeneity of this receptor family, for example a peripheral Y2 receptor. However, since many studies have been carried out with different peptide analogs and radioligands in different species, there is substantial confusion regarding the pharmacological profile of the receptors. This may have led to an exaggeration of the potential number of discrete receptors. PMID- 9223223 TI - Maturation of the mammalian dorsal root entry zone--from entry to no entry. AB - Interfaces between glial cell precursors of the PNS and CNS are established early in development and form the sites where sensory axons enter and motor axons exit the developing CNS. The molecular and cellular interactions that lead to the formation of these glial interfaces are only now becoming apparent. New in-vitro techniques are providing clues as to how the maturation of PNS-CNS glial interfaces generates barriers to regenerating axons. PMID- 9223224 TI - Bone morphogenetic proteins in the nervous system. AB - Bone morphogenetic proteins (BMPs) are a rapidly expanding subclass of the transforming growth factor superfamily. BMP ligands and receptor subunits are present throughout neural development within discrete regions of the embryonic brain and within neural crest-derived pre- and post-migratory zones. BMPs initially inhibit the formation of neuroectoderm during gastrulation while, within the neural tube, they act as gradient morphogens to promote the differentiation of dorsal cell types and intermediate cell types throughout co operative signaling. In the peripheral nervous system, BMPs act as instructive signals for neuronal lineage commitment and promote graded stages of neuronal differentiation. By contrast, within the CNS, these same factors promote astroglial lineage elaboration from embryonic subventricular zone progenitor cells, with concurrent suppression of the neuronal or oligodendroglial lineages, or both. In addition, BMPs act on more lineage-restricted embryonic CNS progenitor cells to promote regional neuronal survival and cellular differentiation. Furthermore, these versatile cytokines induce selective apoptosis of discrete rhombencephalic neural crest-associated cellular populations. These observations suggest that the BMPs exhibit a broad range of cellular and context-specific effects during multiple stages of neural development. PMID- 9223222 TI - Nitric oxide, the enigmatic neuronal messenger: its role in synaptic plasticity. AB - The discovery of the intercellular messenger nitric oxide (NO) stimulated new concepts of how synaptic plasticity could be induced in the nervous system. While initial reports found evidence that NO is of importance for the formation of long term potentiation of synaptic transmission (LTP) and spatial learning in rats, later reports failed to confirm these results. Novel approaches such as deletion of the gene that encodes NO synthase in mice showed that the neuronal and the endothelial isoforms are expressed in neurones. Deletion of both isoforms reduced the inducibility of LTP. Furthermore, novel selective inhibitors of NO synthase impaired spatial learning. These results support the hypothesis that NO plays an important role in synaptic transmission and explain some but not all previously contradictory results. PMID- 9223226 TI - Agonist-induced adherence of equine eosinophils to fibronectin. AB - Eosinophils are believed to play an important part in the pathogenesis of equine diseases such as helminth infestation and the allergic skin disease, sweet itch. It has been shown that adherence of human eosinophils to the connective tissue matrix protein fibronectin enhances cell activation and survival time. If adherence causes similar changes in the properties of equine eosinophils, cell induced tissue damage at a site of parasitic infestation or allergic response would be exacerbated. However, investigation of this hypothesis requires identification of mediators that cause equine eosinophil adherence. Since the equivalent recombinant equine proteins were not available, the present study reports the effects of recombinant human (rh) C5a and IL-5 on the adherence of equine peripheral blood eosinophils (EPBEs) to fibronectin in vitro. The effects of LTB4 and PAF on EPBE adherence to fibronectin were also examined and phorbol myristate acetate (PMA) was used as a positive control. PMA caused a dose-related increase in EPBE adherence to fibronectin-coated plastic. In comparison, rh C5a produced a much smaller response which was only evident at the highest dose tested. On the other hand, rhIL-5 induced a small, but significant dose-related increase in EPBE adherence. Moreover, this response was in part dependent on the beta 1 integrin Very Late Antigen-4 (VLA4). Since adherence to serum-coated plastic was also increased by IL-5, beta 2 integrins may be activated and/or up regulated on EPBEs by the cytokine. Neither LTB4 nor PAF caused EPBE adherence to fibronectin but prior incubation with these mediators increased the response of cells to IL-5. There were no differences between the responses of EPBEs isolated from horses with clinical signs of sweet itch and normal animals. Thus, whilst up regulation of IL-5-induced adherence may occur locally in tissues in vivo, it does not appear to take place in the circulation. Finally, C5a, PAF and LTB4, but not IL-5, caused equine neutrophil adherence to fibronectin demonstrating the different responses of granulocytes to these mediators. The results obtained in the present study have shown that mediators which may be released at sites of inflammatory or allergic reactions can induce or enhance eosinophil adherence to tissue matrix protein. Thus, these mediators can now be used in future studies to determine if cell adherence may alter eosinophil activation or survival time. PMID- 9223225 TI - Cytokine RNA expression in an equine CD4+ subset differentiated by expression of a novel 46-kDa surface protein. AB - Two monoclonal antibodies (MAb), HB65A (IgG2a) and HB86A (IgGI), recognize a unique cell surface molecule on equine T-lymphocytes. The molecule, designated EqWC4, identified by these MAbs is present on a subpopulation of CD4+ equine lymphocytes (6.3-10.2% of Arabian lymphocytes CD4+ WC4+) and a smaller population of CD8+ lymphocytes (0.5% to 1.2% of Arabian lymphocytes CD8+ WC4+). EqWC4 is absent from B-lymphocytes, granulocytes, and macrophages. Both MAbs bound to a 46 kDa protein following immunoprecipitation reactions with lysates of surface labeled thymocytes. Immunoaffinity purification using HB65A yielded two molecules of 46 kDa and 52 kDa under reducing conditions and a third 92-kDa molecule was present in nonreduced conditions. Activation by mitogen did not increase expression of EqWC4 on equine lymphocytes. Lymphocytes from Arabian, Pony, and Thoroughbred breeds showed a common distribution of EqWC4 among leukocytes. However, there were significantly fewer Pony lymphocytes bound to HB65A and HB86A when compared to Arabian and Thoroughbred breeds. Using reverse transcriptase polymerase chain reaction (RT-PCR), magnetically enriched populations (to 80% of cells isolated) of EqWC4+ lymphocytes expressed a cytokine RNA profile dominated by -interleukin2 (IL-2) and interferon-gamma (IFN-gamma) for unstimulated cells. Upon mitogen stimulation, IL-4 was also expressed at low levels while the IL-2 levels decreased and the IFN-gamma levels increased relative to unstimulated cells. EqWC4 is similar to CD28 in molecular weight and its formation of dimers and could therefore be the equine orthologue. However, because of the differences in CD28 expression, EqWC4 probably represents a previously uncharacterized equine lymphocyte marker. PMID- 9223227 TI - Molecular cloning and functional expression of equine interleukin-1 receptor antagonist. AB - Equine interleukin-1 receptor antagonist (IL-1ra) was molecularly cloned to establish a basis for cytokine therapy of acute and chronic inflammatory diseases in the horse. cDNA clones encoding the whole coding sequence of equine IL-1ra were isolated from equine peripheral blood mononuclear cells (PBMC) that had been stimulated with lipopolysaccharide (LPS). The equine IL-1ra cDNA obtained in this study contained an open reading frame encoding 177 amino acid residues. The predicted amino acid sequence of equine IL-1ra shared 75.7, 75.3 and 76.3% similarity with sequences of human, murine and rabbit IL-1ras, respectively. An N glycosylation site and five cysteine residues conserved in human, murine and rabbit IL-1ras were also found at the corresponding positions in equine IL-1ra. Recombinant glutathione S-transferase (GST)-equine IL-1ra fusion protein produced by Escherichia coli was purified. This protein was shown to inhibit the cytostatic or cytotoxic activity of IL-1 on A375S2 cells, indicating that the equine IL-1ra cDNA obtained in this study encodes biologically active equine IL 1ra. PMID- 9223228 TI - Functional characterization of equine neutrophils in response to calcium ionophore A23187 and phorbol myristate acetate ex vivo. AB - Equine neutrophils (PMN) play a critical role in inflammatory processes in horses. The objective of this study was to characterize equine PMN function ex vivo following stimulation with calcium ionophore A23187 (A23187) and phorbol myristate acetate (PMA). These stimulants trigger different branches of the PMN activation process that occurs in vivo. Equine PMN were isolated from the whole blood of six clinically normal geldings using a one-step discontinuous Percoll gradient technique. Neutrophil aggregation, degranulation, and superoxide anion production were evaluated in assay systems which had previously been established to quantitate PMN function. Dose-response curves for A23187 and PMA were derived for the three functions. Results indicate that equine PMN aggregation and superoxide anion production are more responsive to activation by PMA as the maximum change in percent transmittance and maximum nanomoles of superoxide anion produced following PMA stimulation (60.8% and 10.4 nmol per 10(6) cells, respectively) were greater than those values stimulated by A23187 (41.5% and 5.2 nmol per 10(6) cells, respectively). However, degranulation was found to be more responsive to A23187 stimulation (maximum percent degranulation: 56.1%) than to PMA stimulation (maximum percent degranulation: 30.7%). Dose-response curves following A23187 and PMA stimulation revealed that superoxide anion production had the lowest threshold concentration among the three functions. Degranulation had the highest threshold concentration among the three functions for both stimulants. Results indicate that equine PMN functions differ in their dependence on second messengers in the activation pathway. These functions also occur in a dose-dependent manner and differ in the threshold concentrations required for their stimulation. PMID- 9223229 TI - Genomic structure of the bovine mb-1 gene encoding the Ig-alpha subunit of the B cell antigen receptor complex. AB - The B cell antigen receptor, (BCR) comprises surface immunoglobulin and disulfide bonded heterodimer of Ig-alpha and Ig-beta chains, which are the products of the mb-1 and B29 genes, respectively. In this study, we describe the isolation and analysis of a 6.2-kb genomic DNA clone containing bovine mb-1 gene encoding Ig alpha. Sequence data revealed that the bovine mb-1 gene is composed of five exons and four introns, and that its overall structure is very similar to those of murine and human genes. The 5' upstream region of the bovine mb-1 gene contained potential protein binding motifs of transcription factors including EBF, Sp1, NF kappa B, MUF/Ets-1 and AP 2. As with the murine and human mb-1 genes, the 5' region of the bovine mb-1 gene lacked a TATA box. The present study will be useful for understanding the regulated expression of the bovine mb-1 gene at different stages of development and activation as well as in bovine leukemia virus infection. PMID- 9223230 TI - Simultaneous flow cytometric measurement of phagocytotic and oxidative burst activity of polymorphonuclear leukocytes in whole bovine blood. AB - Polymorphonuclear neutrophils (PMN) are important in host defence against bacterial infection in the bovine mammary gland and techniques are needed to evaluate their functional activities. A rapid and sensitive two-color flow cytometric method for simultaneous measurement of phagocytosis rate and oxidative burst activity of bovine PMN in small blood samples is described. The method utilizes the oxidation of intracellular dihydrorhodamine 123 to green fluorescent rhodamine 123 by oxidative burst products that were generated by incubating the PMNs with red fluorescent propidium iodide labeled Staphylococcus aureus. Phagocytosis and oxidative burst both increased with time of incubation and with increasing bacteria concentration. A 20 min phagocytosis time and a ratio of 25 bacteria to one cell were considered optimal for assaying bovine blood PMN activity. To further evaluate the proposed two-color flow cytometric method, blood samples were treated with factors known to interfere with phagocytosis and oxidative burst metabolism of human neutrophils. Incubation of whole bovine blood with cytochalasin B at 10 micrograms ml-1 resulted in an approximate 70% decrease in the phagocytosis rate with a concommitant decrease in oxidative burst activity. Staurosporine (2 micrograms ml-1) inhibited bovine neutrophil oxidative burst formation for 95% while the phagocytosis was unaffected. PMNs in whole blood samples of ten cows differed significantly in their ability to phagocytose Staphylococcus aureus and to produce reactive oxygen products. However, only a weak correlation was detected between the burst activity:phagocytosis ratio of ten individual cows as indicated by the ROD/PI index. PMID- 9223231 TI - Influence of antimicrobial agents on bactericidal activity of bovine milk polymorphonuclear leukocytes. AB - The influence of nine commonly used antibiotics on the respiratory burst activity of bovine milk polymorphonuclear leukocytes (PMNL) of high yielding cows was studied in vitro. Cellular oxidative activity was quantitated after preincubation with drugs at different concentrations and assayed by a PMA (12,13-phorbol myristate acetate)-induced luminol-enhanced chemiluminescence (CL) technique. All antibiotics except sulphadiazine and enrofloxacin decreased CL at the highest concentration. Enrofloxacin significantly increased CL. Oxytetracycline inhibited CL even at low doses. The decreased CL with danofloxacin and oxytetracycline was mainly induced by their color, which caused absorption of the blue light emitted by luminol. Production of superoxide radicals measured by the cytochrome c reduction assay was lowered by danofloxacin, penicillin and chloramphenicol. The decreased CL with ceftiofur was due to inhibition of myeloperoxidase (MPO) activity and to scavenging of reactive oxygen species. Interference with the MPO H2O2-halide system was also observed with spiramycin, erythromycin and oxytetracycline, while the latter was also observed with penicillin. The stimulatory effect of enrofloxacin might be due to an improvement of the penetration of luminol into the PMNL or to a stimulation of the production of H2O2. Potentiation of the action of PMA by changing the ratio between bound and free intracellular Ca2+ might also be involved. Our results suggest that many antibiotics may affect neutrophil function at concentrations that may be found in milk immediately after intramammary treatment or at concentrations higher than those found in milk after intramammary treatment. PMID- 9223232 TI - Cell-mediated immune response and IL-2 production in white-tailed deer experimentally infected with hemorrhagic disease viruses. AB - Hemorrhagic disease, caused by various serotypes of two closely related orbivirus serogroups, the epizootic hemorrhagic disease viruses (EHDV) and the bluetongue viruses (BTV), is a major cause of morbidity and mortality in white-tailed deer (WTD) in the United States. Despite the importance of hemorrhagic disease in WTD, little is known about host defense mechanisms triggered by infection with either causative virus or how that immune response is modulated by challenge with closely related orbiviruses, as can occur under natural conditions. Initial experimental data from our laboratory showed WTD infected with EHDV serotype 2 (EHDV-2) had responded serologically but often became lymphopenic and had a reduced lymphocyte proliferative response in vitro to T-cell mitogens, suggesting possible suppression of cell-mediated immunity. The primary objective of this study was to more closely examine cell-mediated immunity of WTD when experimentally infected with EHDV-2 and subsequently challenged with BTV serotype 10 (BTV-10). The cell-mediated response was evaluated via in vitro lymphocyte proliferation and interleukin-2 (IL-2) production assays, and in vivo delayed type hypersensitivity tests. Deer infected with either EHDV-2 or BTV-10 responded similarly in all assays. Infected deer had decreased lymphocyte counts between post-infection days (PID) 6 and 10, with concurrent diminished lymphocytic response to concanavalin A in lymphocyte proliferation assays and phytohemagglutinin in delayed, type hypersensitivity tests. However, IL-2 production by peripheral blood lymphocytes of infected deer was comparable with that of non-infected control deer as measured using a IL-2-dependent bovine cell line (BT2). This suppression of T-cell proliferation, but not IL-2 production suggests selective inhibition of T-cells probably via altered signal transduction for either expression of the IL-2 receptor or for IL-2 receptor signal-induced T cell proliferation. PMID- 9223233 TI - Nitric oxide production following in vitro stimulation of ovine pulmonary alveolar macrophages. AB - Production of inducible nitric oxide (NO) as measured by nitrite in supernatant from ovine pulmonary alveolar macrophage (PAM) cultures was assessed following stimulation of PAM with live cells and supernatants from Corynebacterium pseudotuberculosis and Pasteurella haemolytica cultures; purified bacterial lipopolysaccharide derived from both Escherichia coli and Pasteurella haemolytica alone and in combination with interferon-gamma or lymphocyte conditioned medium; or ovine lentivirus. PAM cultured ex vivo with no further stimulation for 24 h, 48 h or 72 h, produced low concentrations of NO that was not substantially increased following co-culture by the various additives. Assessment of NO production in PAM cultures containing P. haemolytica or supernatant from P. haemolytica cultures was complicated by production of high levels of nitrite in the bacterial cultures. Species differences in inducible NO production may affect the efficacy of clearance of bacterial infections and be responsible for inter host differences in disease expression following infection by intracellular pathogens. PMID- 9223234 TI - Excretion of ammonia by Lucilia cuprina larvae suppresses immunity in sheep. AB - Attempts to immunise sheep against natural infestations by Lucilia cuprina larvae have not been effective. Yet it is known that the larvae excrete the immunosuppressant ammonium bicarbonate. The effect of larval ammonium and nonionic ammonia on immunopathobiology was evaluated in 12 infested sheep. The concentration of ammonium in veins draining infested sites was measured in another group of four sheep. Mean jugular unionized ammonia concentration increased 3.5 to 5.6 times above pre-infested control levels. Mean venous ammonium concentrations draining infested sites were 13 times higher than pre infested jugular or carotid levels. Increases in jugular ammonia concentrations correlated with increased number of larvae, area of infestation, earlier death, neutropenia, eosinopenia, lymphocytopenia, large declines in serum globulins and zinc, and large rises in toxic neutrophils. The high concentrations of toxic unionized ammonia in blood directly permanently damaged neutrophils and lymphocytes and depressed serum globulin production. The results show that the ammonium from the excreta of larvae of L. cuprina may be highly immunosuppressive. PMID- 9223235 TI - Epidermal immunocompetence in canine leishmaniasis. AB - The antigen-presenting cell function of the epidermis was investigated immunocytochemically in 16 dogs with different types of skin lesions induced by Leishmania infection. The degree of epidermal immunocompetence was evaluated according to the presence of Langerhans cells (LC) and keratinocytes expressing class II major histocompatibility complex (MHC II) molecules on the one hand, and the relative numbers of macrophages, T cells, and parasites in the dermis on the other, as described for human cutaneous leishmaniasis. In alopecic dermatitis, appropriate numbers of LC and MHC II-positive keratinocytes were shown to be associated with a mild T cell infiltration without significant numbers of parasites. By contrast, when the epidermis lacked antigen-presenting cells, as occurred in nodular lesions, macrophages and parasites massively infiltrated the dermis. Ulcerative lesions showed intermediate patterns of inflammation. These results suggest that dogs with alopecic dermatitis develop an effective control of the infection, whereas those with a generalized nodular disease mount an impaired immune response against the parasite. Skin lesions in dogs infected by Leishmania might not only have a prognostic value, but also represent a suitable model to study the natural course of human cutaneous leishmaniasis. PMID- 9223236 TI - Postnatal development of B lymphocytes and immunoglobulin-containing plasma cells in the chicken oviduct: studies on cellular distribution and influence of sex hormones. AB - Postnatal development of B lymphocytes and plasma cells containing different classes of immunoglobulins (IgG, IgA, and IgM) was immunohistochemically studied in the oviduct of the Dekalb strain of the White Leghorn chicken. B lymphocytes first appeared in the lamina propria of the chicken oviduct at 5 weeks of age. Their frequency of occurrence peaked at 15 weeks from the infundibulum to the uterus (glandular part), while in the vagina (aglandular part) it did so at 21 weeks. Intraepithelial B lymphocytes were very rare and exclusively located in the vagina at 19 and 21 weeks. Plasma cells first appeared in the lamina propria of the oviduct at 11 weeks of age, and this frequency peaked at 32 weeks. IgG containing plasma cells were most numerous in the glandular part, whereas in the aglandular part IgA and IgM cells were more numerous than IgG cells. When 7-day old-chickens were treated with sex hormones, B lymphocytes and plasma cells appeared 12 h and 5 days after the hormone injection, respectively. Their frequency of occurrence was statistically higher in diethylstibestrol (DES) treated chickens than in DES plus progesterone-treated chickens. This suggests that the postnatal development of B lymphocytes and plasma cells in the oviduct of the chicken is correlated to estrogen secretion. PMID- 9223237 TI - Effect of permeabilization on peripheral blood lymphocytes of bovine leukemia virus (BLV)-infected cattle. AB - Cell surface proteins serve as markers for immunophenotypic characterization of lymphocyte subsets by appropriate monoclonal antibodies and fluorescence activated cell sorter (FACS) analysis. By the same method, internal antigens or those that are only partially expressed on the cell surface can be determined after permeabilization of the cells. Peripheral blood lymphocytes obtained from bovine leukemia virus (BLV)-infected cattle and from BLV-free cattle were permeabilized and several lymphocyte populations were examined. BoCD4, BoCD8 and three CD4 CD8-T-cell subsets retained their original frequencies after permeabilization in both groups of animals. The recognition of the B-B2 lymphocyte molecule was only partially expressed on the cell surface of intact lymphocytes and was further revealed on permeabilization. The frequency of permeabilized, but not intact, cells stained with this mAb was significantly higher for BLV-infected cattle than for BLV-free animals (P = 0.006). Reactivities of an anti-heat shock protein (Hsp) 70 were measured before and after permeabilization of PBLs. Similar increased cell frequencies were obtained for both groups of bovines. These data indicate that flow cytometry studies should be conducted on both permeabilized and intact cells for a better assessment of protein expression on the cell surface, as well as in the cytoplasm. PMID- 9223238 TI - Lymphocyte subset abnormalities in canine leishmaniasis. AB - Peripheral blood lymphocyte subpopulations were studied in 42 Leishmania infantum infected dogs using flow cytometry. Twenty-two healthy dogs were used as a control group. Analysis of the B-cell populations showed a reduction in the number of CD21+ cells in all the infected dogs. On the other hand, the disease was found to be associated with a striking decrease in the number of CD21+ cells and of T-lymphocyte CD4+ cells in comparison with asymptomatic dogs and with healthy dogs. This study suggests that the dysimmunity which is observed with leishmaniasis may be linked to a reduced number of T-lymphocyte CD4+ cells. PMID- 9223239 TI - Effect of storage of chicken and turkey blood on the lymphocyte responses to concanavalin A and pokeweed mitogen. AB - The effect of storage of whole blood of chicken and turkeys on the mitogen responses of lymphocytes to concanavalin A (con A) and pokeweed mitogen was investigated. It was found that, despite slight differences in optimum storage conditions with respect to mitogens, species and age of birds, blood could be stored for 24 h at 4 degrees C without significant reduction in the lymphoproliferative responses. However, even during the initial 24 h storage of blood at 4 degrees C, the levels of reduction in stimulation index (SI) values following con A stimulation, ranged from 27% for 3-4 week-old turkeys to 60% for 3-4 week-old white leghorn chickens. PMID- 9223241 TI - Procaine-induced maturation of Xenopus oocytes is mediated by a transient activation of M-phase promoting factor. AB - We have recently shown that the incubation of Xenopus laevis oocytes in procaine containing solutions induced germinal vesicle breakdown without white spot formation and, in some cases, with the appearance of spindle and chromosomes in the cytoplasm. The present study was performed to determine whether M-phase promoting factor was involved in this unusual maturation. Procaine failed to induce maturation in the presence of 6-dimethylamino purine or roscovitine, which are both known to inhibit p34cdc2 kinase. Histone H1 kinase activity was detected in procaine-treated oocytes but it was always lower than in progesterone-treated controls. A shift in p34cdc2 was observed in oocytes that had been exposed to procaine for 16 h, but it was not detected in those exposed for 24 h. Finally, cytoplasm transfer experiments demonstrated that the maturation promoting activity that occurred in oocytes incubated in procaine for 16 h could induce maturation of recipient stage VI oocytes. This transferable activity was weaker than that from progesterone-treated controls since only 30% of the recipients underwent germinal vesicle breakdown and only a few spindles were observed, which were not always correctly located. Taken together these results demonstrate that M-phase promoting factor is involved in the procaine maturing effect despite some differences compared with progesterone-treated oocytes which might explain the particular type of maturation induced by this substance. The discovery of the mechanisms by which procaine is able to activate M-phase promoting factor might now help in the understanding of some steps in progesterone-induced maturation that have still to be elucidated. PMID- 9223240 TI - Intracellular ion concentrations and their maintenance by Na+/K(+)-ATPase in preimplantation mouse embryos. AB - We have measured the amounts of Na+, K+ and C- in preimplantation mouse embryos (1-cell, 2-cell and morula) using electron probe X-ray microanalysis. The levels of these ions do not vary much over this period, and are approximately the same as those found in other mammalian cells, contrary to previous reports. We have confirmed that preimplantation embryos exhibit Na+/K(+)-ATPase activity at all stages examined, and have shown that the ATPase maintains high K+/Na+ ratios (12 16) in all these embryonic stages, comparable to those seen in other healthy cells; this is in contrast to the low ratios reported in earlier work. Inhibition of the Na+/K(+)-ATPase results in the slow exchange of intracellular K+ for extracellular Na+ (half-time approximately 5 h), indicating that Na+/K(+)-ATPase activity maintains steep Na+ and K+ gradients in preimplantation mouse embryos as it does in most other cells. PMID- 9223242 TI - Virus-like particles, bacteria and microsporidia affect spindle-associated membranes in spermatocytes of Lepidoptera species. AB - Larval testes of four Lepidoptera species were examined using electron microscopy. The testes of one species, the Mediterranean mealmoth Ephestia kuehniella (Pyralidae), were devoid of intracellular pathogens and serve as a control. In this species, metaphase spindles of primary spermatocytes showed a thick layer of perispindle membranes. The membranes were structurally very similar to the agranular endoplasmic reticulum. Membranes of this type occurred also at high frequency throughout the spindle matrix. The analysis of larval testes of Pieris brassicae (Pieridae) revealed virus-like particles within spermatocytes. In another species, Philudoria potatoria (Lasiocampidae), the spermatocytes possessed intracellular bacteria. Whereas the pathogens were found within the germ cells in these species, a fourth species, Plutella xylostella (Plutellide), showed microsporidia within somatic cells of the testis sheath. In all the infected animals, the mass of perispindle membranes was reduced in comparison with spermatocytes of E. kuehniella. However, spindle structure appeared regular in the infected animals. This indicates that a thick layer of perispindle membranes is not decisive for spindle assembly and function in male meiosis of Lepidopera. PMID- 9223243 TI - Isolation of viable egg cells of rape (Brassica napus L.). AB - The isolation of viable egg cells of rape (Brassica napus L.) has been achieved from microdissected ovules. The non-gametic cells of the embryo sac, synergids and central cells have also been isolated. Their morphology corresponded to that of these cells in situ, making a discrimination from isolated sporophytic cells possible. Two hours after isolation the egg cells were still viable. Viable egg cells have been reproducibly isolated with a frequency of 25% per dissected ovule. PMID- 9223244 TI - Sperm nuclear envelope: breakdown of intrinsic envelope and de novo formation in hamster oocytes or eggs. AB - During fertilisation of a fully mature oocyte, the sperm intrinsic nuclear envelope (SINE) disappears soon after sperm-oocyte fusion. A new nuclear envelope appears around the decondensed sperm chromatin when the oocyte reaches telophase II. Whether the SINE persists or rapidly disappears after sperm entry into immature oocytes or fertilised eggs has been controversial. Nuclear envelopes have been demonstrated around the sperm chromatin, which cannot be decondensed within the ooplasm of these oocytes or eggs, but whether these envelopes are persisting SINEs or newly formed envelopes has been a point of dispute. To resolve this issue, the fate of the SINEs of hamster sperm nuclei was traced after incorporation into immature oocytes at the germinal vesicle stage (GV oocytes) or fertilised eggs at the pronuclear stage (PN eggs). The SINEs disappeared quickly within these oocytes or eggs, like those within maturing or mature oocytes, suggesting that the envelopes around the sperm chromatin must be newly formed after SINE breakdown. To obtain further evidence, a detergent treated, SINE-free sperm nucleus was injected into a PN egg. A new envelope appeared around the still-condensed or partially decondensed sperm chromatin within 3 h after injection. Thus, disassembly of the SINE within ooplasm, unlike that of nuclear envelopes of other cells at prophase, is independent of the cell cycle stage of the oocyte or egg, whereas the ability of the ooplasm to assemble the new envelope is restricted to certain periods of the cycle, i.e. early prophase and telophase during meiosis and interphase, periods when active M-phase promoting factor (MPF) is absent from the ooplasm. PMID- 9223246 TI - Effects of oviductal fluid and heparin on fertility and characteristics of porcine spermatozoa. AB - The objective of this study was to determine the effects of oviductal fluid and heparin on sperm penetration and the characteristics of spermatozoa. The addition of oviductal fluid and heparin to the fertilisation medium decreased sperm penetration and the mean number of spermatozoa in penetrated eggs. The number of spermatozoa firmly bound to zona pellucida was also decreased in the presence of oviductal fluid and heparin. Chlortetracycline (CTC) fluorescence patterns were used to determine the incidence of capacitation and the acrosome reaction. The proportion of capacitated and acrosome-free spermatozoa increased when spermatozoa were exposed for 1.5 and 3 h to oviductal fluid and heparin. In contrast heparin alone did not increase the number of capacitated spermatozoa at these time points. These results suggest that factor(s) in oviductal secretions reduce polyspermic fertilisation and the number of spermatozoa that will penetrate porcine oocytes. The reduction of polyspermic penetration by oviductal secretions may be due to a reduced number of spermatozoa in the fertilisation medium with an intact acrosome. PMID- 9223247 TI - Stage-specific requirement of phosphate for development of rat 1-cell embryos in a chemically defined medium. AB - Rat 1-cell embryos, recovered from naturally mated females, were cultured in a chemically defined medium (mR1ECM). When examined after 24, 56, 64 and 80 h of culture, embryos developed to the 2-cell (100%), 4-cell (93%), 4-cell to 8-cell (97%) and > or = 8-cell (95%) stages, respectively. When 0.4 M phosphate (NaH2PO4) was added to the medium after 0, 24, 56 and 64 h of culture, percentages (0-67%) of embryos that developed to the blastocyst stage after 115 h of culture were lower than that (86%) in the medium without phosphate. However, addition of phosphate after 80 h of culture accelerated blastocyst formation; a significantly higher percentage (94%) of blastocysts was obtained after 110 h of culture in this medium compared with when phosphate was not added (67%). When phosphate was added to the medium after 64 h, almost all (97%) 8-cell embryos developed to the blastocyst stage by 100 h of culture but development of 4-cell to 7-cell embryos was inhibited (22-63%). Acceleration of blastocyst formation was caused by addition of phosphate rather than by the exchange of the medium. The stimulatory effect of phosphate on embryo development was observed at concentrations of 0.1-1.2 mM. The mean numbers of cells (54.5-60.9 cells) in blastocysts examined after 115 h of culture were increased by the addition of 0.4 1.6 mM phosphate at 80 h as compared with blastocysts from cultures without phosphate (46.6 cells). PMID- 9223245 TI - Ultrastructural and immunocytochemical analysis of diploid germ cells isolated from fetal rabbit gonads. AB - Germ cells were isolated from rabbit fetal gonads between 18 and 22 days post coitum and examined morphologically, ultrastructurally and for immunocytochemical and cytochemical characteristics. Observations were compared with the information available from the corresponding cells of other mammalian species. The general morphology and ultrastructure of healthy isolated rabbit fetal germ cells were found to be very similar to those of the rabbit and mouse diploid germ cells in situ. Moreover, rabbit fetal germ cells shared common immunocytochemical characteristics with mouse undifferentiated embryonic stem cells or embryonic carcinoma cells, such as the presence of TEC-1 (SSEA-1) antigens, a peripheral network of F-actin, the absence of cytokeratins 8/18 and lamins A/C and an alkaline phosphatase activity. No difference between the sexes was observed. Morphological and physiological similarities with the migrating and cultured primordial germ cells of the mouse also suggest that diploid rabbit germ cells would be good candidates for deriving pluripotential embryonic germ cells (EG cells) if favourable culture conditions could be found. In conclusion, the rabbit may be a suitable model for investigations on EG cells in domestic mammals with delayed meiosis. PMID- 9223248 TI - Calcium- and meiotic-spindle-independent activation of pig oocytes by the inhibition of staurosporine-sensitive protein kinases. AB - The dependence of pig oocyte activation (both nuclear activation and cortical granule exocytosis) induced by staurosporine on intracellular Ca2+ rise and spindle assembly was studied. Nuclear activation was evaluated by pronuclear (PN) formation, cleavage and their developmental ability, and cortical granule (CG) exocytosis was assessed by electron microscopy and laser confocal microscopy of oocytes labelled with fluorescein isothiocyanate-peanut agglutinin. Exposure of pig oocytes of 0.3 and 3 microM protein kinase inhibitor staurosporine for 30 min resulted in the nuclear activation in 71.8% and 85.7% of the oocytes, respectively. The pronuclei in activated oocytes contained several compact nucleoli. When the cleaved 2-cell oocytes were further cultured in vitro, 93.5% developed beyond the 4-cell stage, and 12.9% developed to the morula stage after 4 days of culture. Of the oocytes treated with 3 microM staurosporine, 62.5% and 9.4% released their CGs partially and completely, respectively. The nuclear activation induced by staurosporine was overcome by the prior treatment of oocytes with okadaic acid, resulting in only 33.3% of the oocytes undergoing nuclear activation. However, when oocytes were exposed first to 1,2-bis(O aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (acetoxymethanal ester), a cell permeate calcium chelator, or Colcemid, a meiotic spindle disrupter, and then to staurosporine, nuclear activation was observed in 74.2% and 82.3% of the oocytes, respectively. These data were the same as those in oocytes treated only with staurosporine (85.7%). The present study indicates that pig oocytes can be activated by the inhibition of staurosporine-sensitive protein kinase(s), and that this activation is dependent upon mitogen-activated protein kinase but independent of the intracellular Ca2+ rise and spindle integrity. PMID- 9223249 TI - Microinjection of anti-alpha-tubulin antibody (DM1A) inhibits progesterone induced meiotic maturation and deranges the microtubule array in follicle enclosed oocytes of the frog, Rana pipiens. AB - Microinjection of anti-alpha-tubulin (Dm1A) inhibited progesterone-induced meiotic maturation in large follicle-enclosed oocytes of the frog, Rana pipiens. DM1A (46 nl; 10 mg/ml) injection significantly increased the ED50 value for progesterone as determined by germinal vesicle dissolution (GVD) bioassay. By contrast, low doses of microinjected DM1A (46 nl; 2.5 mg/ml), anti-actin (clone KJ43A), anti-cytokeratin (C-11), anti-intermediate filament antibody (IFA), generic IgG (46 nl; 20 mg/ml) or sodium azide (46 nl; 1 mg/ml), an antibody preservative, were without inhibitory effect in this bioassay. Microinjected, affinity-purified DM1A (46 nl; 7.5 mg/ml) was also inhibitory, but preabsorption with pure tubulin prior to injection significantly reduced the inhibitory effect. DM1A injection had no effect on centrifugation-induced germinal vesicle migration (GVM). Previous work indicated that drugs (e.g. demecolcine and nocodazole), which destabilise microtubules, enhance both centrifugation-induced GVM and progesterone-induced GVD in Rana oocytes. Taking these results together, it is suggested that DM1A injection may have differential effects on microtubules in this cell. Thus, while the majority of microtubules were apparently depolymerised by DM1A (46 nl; 10 mg/ml) injection, a small subpopulation appeared to be stabilised as bundles. Confocal immunofluorescence microscopy of follicle enclosed oocytes after DM1A injection revealed a major loss of microtubules throughout the cell; however, apparent sparse bundles of microtubules arranged in an approximately 600 microns shell were associated with the injectate region 24 h post-injection. By contrast, control follicle-enclosed oocytes topically labelled with DM1A post-fixation had extensive microtubule arrays similar to those previously reported in Xenopus oocytes. Intracellular recording after DM1A injection and progesterone treatment yielded an intermediate membrane potential (Vm = -31.8 mV) compared with control (immature) DM1A-injected cells (Vm = -44.7 mV) or potassium balanced salt solution (KBS)-injected cells matured with progesterone (Vm = -13.9 mV). These results suggest that DM1A injection does not completely inhibit electrophysiological changes initiated by progesterone. Working hypotheses are proposed that suggest a role for microtubules in the action of progesterone which normally lifts the prophase I block in the Rana follicle-enclosed oocyte. PMID- 9223250 TI - Genetics and the origin of species: an introduction. PMID- 9223251 TI - Origin of genes. AB - We discuss two tests of the hypothesis that the first genes were assembled from exons. The hypothesis of exon shuffling in the progenote predicts that intron phases will be correlated so that exons will be an integer number of codons and predicts that the exons will be correlated with compact regions of polypeptide chain. These predictions have been tested on ancient conserved proteins (proteins without introns in prokaryotes but with introns in eukaryotes) and hold with high statistical significance. We conclude that introns are correlated with compact features of proteins 15-, 22-, or 30-amino acid residues long, as was predicted by "The Exon Theory of Genes." PMID- 9223253 TI - Long term trends in the evolution of H(3) HA1 human influenza type A. AB - We have studied the HA1 domain of 254 human influenza A(H3N2) virus genes for clues that might help identify characteristics of hemagglutinins (HAs) of circulating strains that are predictive of that strain's epidemic potential. Our preliminary findings include the following. (i) The most parsimonious tree found requires 1,260 substitutions of which 712 are silent and 548 are replacement substitutions. (ii) The HA1 portion of the HA gene is evolving at a rate of 5.7 nucleotide substitutions/year or 5.7 x 10(-3) substitutions/site per year. (iii) The replacement substitutions are distributed randomly across the three positions of the codon when allowance is made for the number of ways each codon can change the encoded amino acid. (iv) The replacement substitutions are not distributed randomly over the branches of the tree, there being 2.2 times more changes per tip branch than for non-tip branches. This result is independent of how the virus was amplified (egg grown or kidney cell grown) prior to sequencing or if sequencing was carried out directly on the original clinical specimen by PCR. (v) These excess changes on the tip branches are probably the result of a bias in the choice of strains to sequence and the detection of deleterious mutations that had not yet been removed by negative selection. (vi) There are six hypervariable codons accumulating replacement substitutions at an average rate that is 7.2 times that of the other varied codons. (vii) The number of variable codons in the trunk branches (the winners of the competitive race against the immune system) is 47 +/- 5, significantly fewer than in the twigs (90 +/- 7), which in turn is significantly fewer variable codons than in tip branches (175 +/- 8). (viii) A minimum of one of every 12 branches has nodes at opposite ends representing viruses that reside on different continents. This is, however, no more than would be expected if one were to randomly reassign the continent of origin of the isolates. (ix) Of 99 codons with at least four mutations, 31 have ratios of non silent to silent changes with probabilities less than 0.05 of occurring by chance, and 14 of those have probabilities <0.005. These observations strongly support positive Darwinian selection. We suggest that the small number of variable positions along the successful trunk lineage, together with knowledge of the codons that have shown positive selection, may provide clues that permit an improved prediction of which strains will cause epidemics and therefore should be used for vaccine production. PMID- 9223254 TI - Genes, peoples, and languages. AB - The genetic history of a group of populations is usually analyzed by reconstructing a tree of their origins. Reliability of the reconstruction depends on the validity of the hypothesis that genetic differentiation of the populations is mostly due to population fissions followed by independent evolution. If necessary, adjustment for major population admixtures can be made. Dating the fissions requires comparisons with paleoanthropological and paleontological dates, which are few and uncertain. A method of absolute genetic dating recently introduced uses mutation rates as molecular clocks; it was applied to human evolution using microsatellites, which have a sufficiently high mutation rate. Results are comparable with those of other methods and agree with a recent expansion of modern humans from Africa. An alternative method of analysis, useful when there is adequate geographic coverage of regions, is the geographic study of frequencies of alleles or haplotypes. As in the case of trees, it is necessary to summarize data from many loci for conclusions to be acceptable. Results must be independent from the loci used. Multivariate analyses like principal components or multidimensional scaling reveal a number of hidden patterns and evaluate their relative importance. Most patterns found in the analysis of human living populations are likely to be consequences of demographic expansions, determined by technological developments affecting food availability, transportation, or military power. During such expansions, both genes and languages are spread to potentially vast areas. In principle, this tends to create a correlation between the respective evolutionary trees. The correlation is usually positive and often remarkably high. It can be decreased or hidden by phenomena of language replacement and also of gene replacement, usually partial, due to gene flow. PMID- 9223252 TI - Transposable elements as sources of variation in animals and plants. AB - A tremendous wealth of data is accumulating on the variety and distribution of transposable elements (TEs) in natural populations. There is little doubt that TEs provide new genetic variation on a scale, and with a degree of sophistication, previously unimagined. There are many examples of mutations and other types of genetic variation associated with the activity of mobile elements. Mutant phenotypes range from subtle changes in tissue specificity to dramatic alterations in the development and organization of tissues and organs. Such changes can occur because of insertions in coding regions, but the more sophisticated TE-mediated changes are more often the result of insertions into 5' flanking regions and introns. Here, TE-induced variation is viewed from three evolutionary perspectives that are not mutually exclusive. First, variation resulting from the intrinsic parasitic nature of TE activity is examined. Second, we describe possible coadaptations between elements and their hosts that appear to have evolved because of selection to reduce the deleterious effects of new insertions on host fitness. Finally, some possible cases are explored in which the capacity of TEs to generate variation has been exploited by their hosts. The number of well documented cases in which element sequences appear to confer useful traits on the host, although small, is growing rapidly. PMID- 9223255 TI - DNA variation at the Sod locus of Drosophila melanogaster: an unfolding story of natural selection. AB - Patterns of variation at the Sod locus of Drosophila melanogaster suggest that the protein polymorphism at this locus has very recently arisen. In addition, it appears that a previously rare DNA variant has been recently and rapidly driven to intermediate frequency. From the size of the region (>20 kb) that has been swept along with this rare variant, and patterns of linkage disequilibrium in the region, it is inferred that strength of selection was large (s > 0.01) and that the sweep occurred more than 25,000 generations ago. In addition, there are striking similarities to patterns of variation observed at the Est6 and Est-P loci, which are located approximately 1,000 kb from Sod. PMID- 9223256 TI - Neutral behavior of shared polymorphism. AB - Several cases have been described in the literature where genetic polymorphism appears to be shared between a pair of species. Here we examine the distribution of times to random loss of shared polymorphism in the context of the neutral Wright-Fisher model. Order statistics are used to obtain the distribution of times to loss of a shared polymorphism based on Kimura's solution to the diffusion approximation of the Wright-Fisher model. In a single species, the expected absorption time for a neutral allele having an initial allele frequency of 1/2 is 2.77 N generations. If two species initially share a polymorphism, that shared polymorphism is lost as soon as either of two species undergoes fixation. The loss of a shared polymorphism thus occurs sooner than loss of polymorphism in a single species and has an expected time of 1.7 N generations. Molecular sequences of genes with shared polymorphism may be characterized by the count of the number of sites that segregate in both species for the same nucleotides (or amino acids). The distribution of the expected numbers of these shared polymorphic sites also is obtained. Shared polymorphism appears to be more likely at genetic loci that have an unusually large number of segregating alleles, and the neutral coalescent proves to be very useful in determining the probability of shared allelic lineages expected by chance. These results are related to examples of shared polymorphism in the literature. PMID- 9223257 TI - Divergent and conserved features in the spatial expression of the Drosophila pseudoobscura esterase-5B gene and the esterase-6 gene of Drosophila melanogaster. AB - The regulatory regions of homologous genes encoding esterase 6 (Est-6) of Drosophila melanogaster and esterase 5B (Est-5B) of Drosophila pseudoobscura show very little similarity. We have undertaken a comparative study of the pattern of expression directed by the Est-5B and Est-6 5'-flanking DNA to attempt to reveal conserved elements regulating tissue-specific expression in adults. Esterase regulatory sequences were linked to a lacZ reporter gene and transformed into D. melanogaster embryos. Est-5B, 5' upstream elements, give rise to a beta galactosidase expression pattern that coincides with the wild-type expression of Est-5B in D. pseudoobscura. The expression patterns of the Est-5B/lacZ construct are different from those of a fusion gene containing the upstream region of Est 6. Common sites of expression for both kinds of constructs are the third segment of antenna, the maxillary palps, and salivary glands. In vitro deletion mutagenesis has shown that the two genes have a different organization of regulatory elements controlling expression in both the third segment of antenna and maxillary palps. The results suggest that the conservation of the expression pattern in genes that evolved from a common ancestor may not be accompanied by preservation of the corresponding cis-regulatory elements. PMID- 9223258 TI - A demographic approach to selection. AB - The concepts of demography provide a means of combining the ecological approach to population growth with the genetical approach to natural selection. We have utilized the demographic theory of natural selection developed by Norton and Charlesworth to analyze life history schedules of births and deaths for populations of genotypes in Drosophila pseudoobscura. Our populations illustrate a stable genetic equilibrium, an unstable genetic equilibrium, and a case of no equilibrium. We have estimated population growth rates and Darwinian fitnesses of the genotypes and have explored the role of population growth in determining natural selection. The age-specific rates of births and deaths provide insights into components of selection. Both viability and fertility are important components in our populations. PMID- 9223259 TI - Phylogenetics and the origin of species. AB - A recent criticism that the biological species concept (BSC) unduly neglects phylogeny is examined under a novel modification of coalescent theory that considers multiple, sex-defined genealogical pathways through sexual organismal pedigrees. A competing phylogenetic species concept (PSC) also is evaluated from this vantage. Two analytical approaches are employed to capture the composite phylogenetic information contained within the braided assemblages of hereditary pathways of a pedigree: (i) consensus phylogenetic trees across allelic transmission routes and (ii) composite phenograms from quantitative values of organismal coancestry. Outcomes from both approaches demonstrate that the supposed sharp distinction between biological and phylogenetic species concepts is illusory. Historical descent and reproductive ties are related aspects of phylogeny and jointly illuminate biotic discontinuity. PMID- 9223260 TI - Assortative fertilization in Drosophila. AB - The concept of gametic isolation has its origins in the 1937 edition of T. Dobzhansky's Genetics and the Origin of Species. Involving either positive assortative fertilization (as opposed to self-incompatibility) or negative assortative fertilization, it occurs after mating but prior to fertilization. Gametic isolation is generally subsumed under either prezygotic or postmating isolation and thus has not been the subject of extensive investigation. Examples of assortative fertilization in Drosophila are reviewed and compared with those of other organisms. Potential mechanisms leading to assortative fertilization are discussed, as are their evolutionary implications. PMID- 9223261 TI - Incipient species formation in salamanders of the Ensatina complex. AB - The Ensatina eschscholtzii complex of plethodontid salamanders, a well-known "ring species," is thought to illustrate stages in the speciation process. Early research, based on morphology and coloration, has been extended by the incorporation of studies of protein variation and mitochondrial DNA sequences. The new data show that the complex includes a number of geographically and genetically distinct components that are at or near the species level. The complex is old and apparently has undergone instances of range contraction, isolation, differentiation, and then expansion and secondary contact. While the hypothesis that speciation is retarded by gene flow around the ring is not supported by molecular data, the general biogeographical hypothesis is supported. There is evidence of a north to south range expansion along two axes, with secondary contact and completion of the ring in southern California. Current research targets regions once thought to show primary intergradation, but which molecular markers reveal to be zones of secondary contact. Here emphasis is on the subspecies E. e. xanthoptica, which is involved in four distinct secondary contacts in central California. There is evidence of renewed genetic interactions upon recontact, with greater genetic differentiation within xanthoptica than between it and some of the interacting populations. The complex presents a full array of intermediate conditions between well-marked species and geographically variable populations. Geographically differentiated segments represent a diversity of depths of time of isolation and admixture, reflecting the complicated geomorphological history of California. Ensatina illustrates the continuing difficulty in making taxonomic assignments in complexes studied during species formation. PMID- 9223262 TI - Genetics and the origin of bird species. AB - External (environmental) factors affecting the speciation of birds are better known than the internal (genetic) factors. The opposite is true for several groups of invertebrates, Drosophila being the outstanding example. Ideas about the genetics of speciation in general trace back to Dobzhansky who worked with Drosophila. These ideas are an insufficient guide for reconstructing speciation in birds for two main reasons. First, speciation in birds proceeds with the evolution of behavioral barriers to interbreeding; postmating isolation usually evolves much later, perhaps after gene exchange has all but ceased. As a consequence of the slow evolution of postmating isolating factors the scope for reinforcement of premating isolation is small, whereas the opportunity for introgressive hybridization to influence the evolution of diverging species is large. Second, premating isolation may arise from nongenetic, cultural causes; isolation may be affected partly by song, a trait that is culturally inherited through an imprinting-like process in many, but not all, groups of birds. Thus the genetic basis to the origin of bird species is to be sought in the inheritance of adult traits that are subject to natural and sexual selection. Some of the factors involved in premating isolation (plumage, morphology, and behavior) are under single-gene control, most are under polygenic control. The genetic basis of the origin of postmating isolating factors affecting the early development of embryos (viability) and reproductive physiology (sterility) is almost completely unknown. Bird speciation is facilitated by small population size, involves few genetic changes, and occurs relatively rapidly. PMID- 9223263 TI - Vagaries of the molecular clock. AB - The hypothesis of the molecular evolutionary clock asserts that informational macromolecules (i.e., proteins and nucleic acids) evolve at rates that are constant through time and for different lineages. The clock hypothesis has been extremely powerful for determining evolutionary events of the remote past for which the fossil and other evidence is lacking or insufficient. I review the evolution of two genes, Gpdh and Sod. In fruit flies, the encoded glycerol-3 phosphate dehydrogenase (GPDH) protein evolves at a rate of 1.1 x 10(-10) amino acid replacements per site per year when Drosophila species are compared that diverged within the last 55 million years (My), but a much faster rate of approximately 4.5 x 10(-10) replacements per site per year when comparisons are made between mammals ( approximately 70 My) or Dipteran families ( approximately 100 My), animal phyla ( approximately 650 My), or multicellular kingdoms ( approximately 1100 My). The rate of superoxide dismutase (SOD) evolution is very fast between Drosophila species (16.2 x 10(-10) replacements per site per year) and remains the same between mammals (17.2) or Dipteran families (15.9), but it becomes much slower between animal phyla (5.3) and still slower between the three kingdoms (3.3). If we assume a molecular clock and use the Drosophila rate for estimating the divergence of remote organisms, GPDH yields estimates of 2,500 My for the divergence between the animal phyla (occurred approximately 650 My) and 3,990 My for the divergence of the kingdoms (occurred approximately 1,100 My). At the other extreme, SOD yields divergence times of 211 My and 224 My for the animal phyla and the kingdoms, respectively. It remains unsettled how often proteins evolve in such erratic fashion as GPDH and SOD. PMID- 9223264 TI - Evolution of codon usage bias in Drosophila. AB - We first review what is known about patterns of codon usage bias in Drosophila and make the following points: (i) Drosophila genes are as biased or more biased than those in microorganisms. (ii) The level of bias of genes and even the particular pattern of codon bias can remain phylogenetically invariant for very long periods of evolution. (iii) However, some genes, even very tightly linked genes, can change very greatly in codon bias across species. (iv) Generally G and especially C are favored at synonymous sites in biased genes. (v) With the exception of aspartic acid, all amino acids contribute significantly and about equally to the codon usage bias of a gene. (vi) While most individual amino acids that can use G or C at synonymous sites display a preference for C, there are exceptions: valine and leucine, which prefer G. (vii) Finally, smaller genes tend to be more biased than longer genes. We then examine possible causes of these patterns and discount mutation bias on three bases: there is little evidence of regional mutation bias in Drosophila, mutation bias is likely toward A+T (the opposite of codon usage bias), and not all amino acids display the preference for the same nucleotide in the wobble position. Two lines of evidence support a selection hypothesis based on tRNA pools: highly biased genes tend to be highly and/or rapidly expressed, and the preferred codons in highly biased genes optimally bind the most abundant isoaccepting tRNAs. Finally, we examine the effect of bias on DNA evolution and confirm that genes with high codon usage bias have lower rates of synonymous substitution between species than do genes with low codon usage bias. Surprisingly, we find that genes with higher codon usage bias display higher levels of intraspecific synonymous polymorphism. This may be due to opposing effects of recombination. PMID- 9223265 TI - The evolution of plant nuclear genes. AB - We analyze the evolutionary dynamics of three of the best-studied plant nuclear multigene families. The data analyzed derive from the genes that encode the small subunit of ribulose-1,5-bisphosphate carboxylase (rbcS), the gene family that encodes the enzyme chalcone synthase (Chs), and the gene family that encodes alcohol dehydrogenases (Adh). In addition, we consider the limited evolutionary data available on plant transposable elements. New Chs and rbcS genes appear to be recruited at about 10 times the rate estimated for Adh genes, and this is correlated with a much smaller average gene family size for Adh genes. In addition, duplication and divergence in function appears to be relatively common for Chs genes in flowering plant evolution. Analyses of synonymous nucleotide substitution rates for Adh genes in monocots reject a linear relationship with clock time. Replacement substitution rates vary with time in a complex fashion, which suggests that adaptive evolution has played an important role in driving divergence following gene duplication events. Molecular population genetic studies of Adh and Chs genes reveal high levels of molecular diversity within species. These studies also reveal that inter- and intralocus recombination are important forces in the generation allelic novelties. Moreover, illegitimate recombination events appear to be an important factor in transposable element loss in plants. When we consider the recruitment and loss of new gene copies, the generation of allelic diversity within plant species, and ectopic exchange among transposable elements, we conclude that recombination is a pervasive force at all levels of plant evolution. PMID- 9223270 TI - Phosphorylation and subunit organization of axonal neurofilaments determined by scanning transmission electron microscopy. AB - Phosphorylation plays a critical role in controlling the function of cytoskeletal assemblies but no direct method yet exists to measure the phosphorylation state of proteins at the level of individual molecules and assemblies. Herein, we apply scanning transmission electron microscopy in combination with electron energy loss spectroscopy to measure the distributions of mass and phosphorus in neurofilaments (NFs) isolated from the squid giant axon. We find that native squid NFs, in contrast to typical reconstituted intermediate filaments, are a relatively homogeneous population containing only eight coiled-coil dimers per cross section. The measured stoichiometry of approximately 1:1 for light/heavy peptides strongly suggests that squid NFs are composed of heterodimers. Furthermore, each heavy chain of the dimers carries at least 100 phosphate groups and is, therefore, near-maximally phosphorylated. These results also demonstrate that scanning transmission electron microscopy combined with electron energy loss spectroscopy at the nanometer scale is capable of characterizing the level and distribution of phosphorylation in individual mass-mapped protein assemblies. PMID- 9223266 TI - Evolution by the birth-and-death process in multigene families of the vertebrate immune system. AB - Concerted evolution is often invoked to explain the diversity and evolution of the multigene families of major histocompatibility complex (MHC) genes and immunoglobulin (Ig) genes. However, this hypothesis has been controversial because the member genes of these families from the same species are not necessarily more closely related to one another than to the genes from different species. To resolve this controversy, we conducted phylogenetic analyses of several multigene families of the MHC and Ig systems. The results show that the evolutionary pattern of these families is quite different from that of concerted evolution but is in agreement with the birth-and-death model of evolution in which new genes are created by repeated gene duplication and some duplicate genes are maintained in the genome for a long time but others are deleted or become nonfunctional by deleterious mutations. We found little evidence that interlocus gene conversion plays an important role in the evolution of MHC and Ig multigene families. PMID- 9223268 TI - Immunity to retroviral infection: the Friend virus model. AB - Friend virus infection of adult immunocompetent mice is a well established model for studying genetic resistance to infection by an immunosuppressive retrovirus. This paper reviews both the genetics of immune resistance and the types of immune responses required for recovery from infection. Specific major histocompatibility complex (MHC) class I and II alleles are necessary for recovery, as is a non-MHC gene, Rfv-3, which controls virus-specific antibody responses. In concordance with these genetic requirements are immunological requirements for cytotoxic T lymphocyte, T helper, and antibody responses, each of which provides essential nonoverlapping functions. The complexity of responses necessary for recovery from Friend virus infection has implications for both immunotherapies and vaccines. For example, it is shown that successful passive antibody therapy is dependent on MHC type because of the requirement for T cell responses. For vaccines, successful immunization requires priming of both T cell and B cell responses. In vivo depletion experiments demonstrate different requirements for CD8(+) T cells depending on the vaccine used. The implications of these studies for human retroviral diseases are discussed. PMID- 9223271 TI - Inducible gene expression and protein translocation using nontoxic ligands identified by a mammalian three-hybrid screen. AB - The natural product rapamycin has been used to provide temporal and quantitative control of gene expression in animals through its ability to interact with two proteins simultaneously. A shortcoming of this approach is that rapamycin is an inhibitor of cell proliferation, the result of binding to FKBP12-rapamycin associated protein (FRAP). To overcome this limitation, nontoxic derivatives of rapamycin bearing bulky substituents at its C16-position were synthesized, each in a single step. The isosteric isopropoxy and methallyl substituents with the nonnatural C16-configuration abolish both binding to FRAP and inhibition of T cell proliferation. Binding proteins for these derivatives were identified from libraries of cDNAs encoding mutants of the FKBP12-rapamycin-binding (FRB) domain of FRAP by using a mammalian three-hybrid transcription assay. Targeting of the mutations was guided by the structure of the FKBP12-rapamycin-FRB ternary complex. Three compensatory mutations in the FRB domain, all along one face of an alpha-helix in a rapamycin-binding pocket, were identified that together restore binding of the rapamycin derivatives. Using this mutant FRB domain, one of the nontoxic rapamycin derivatives induced targeted gene expression in Jurkat T cells with an EC50 below 10 nM. Another derivative was used to recruit a cytosolic protein to the plasma membrane, mimicking a process involved in many signaling pathways. PMID- 9223273 TI - Helical repeat of DNA in the region of homologous pairing. AB - The process of DNA strand exchange during general genetic recombination is initiated within protein-stabilized synaptic filaments containing homologous regions of interacting DNA molecules. The RecA protein in bacteria and its analogs in eukaryotic organisms start this process by forming helical filamentous complexes on single-stranded or partially single-stranded DNA molecules. These complexes then progressively bind homologous double-stranded DNA molecules so that homologous regions of single- and double-stranded DNA molecules become aligned in register while presumably winding around common axis. The topological assay presented herein allows us to conclude that in synaptic complexes containing homologous single- and double-stranded DNA molecules, all three DNA strands have a helicity of approximately 19 nt per turn. PMID- 9223272 TI - Association of the mammalian helicase MAH with the pre-mRNA splicing complex. AB - Conversion of pre-mRNAs into mature mRNAs includes several consecutive enzymatic modification steps that are carried out in the spliceosomes. Helicases have been shown to contribute to these catalytic processes both in yeast and in mammalian cells. Our results identify the mammalian protein MAH (matrix-associated helicase) as a new helicase present in the spliceosome complex. Sequence comparison describes MAH as the first higher eukaryotic member of the helicase superfamily I, with demonstrated enzymatic activity. Because MAH does not bind small nuclear ribonucleoproteins (snRNPs), it appears to be a non-snRNP binding factor of the splicing complex. In conclusion, our data suggest the involvement of MAH in processing of pre-mRNAs in mammalian cells. PMID- 9223274 TI - Allosteric intermediates indicate R2 is the liganded hemoglobin end state. AB - Hemoglobin has been a long-standing paradigm for understanding protein allostery. Here, the x-ray structures of two chemically crosslinked, fully liganded hemoglobins, alpha2beta82CA82beta and alpha2beta82ND82beta, are described at 2.3 A and 2.6 A resolution, respectively. Strikingly, these crosslinked hemoglobins assume intermediate conformations that lie between those of R and the controversial liganded hemoglobin state R2 rather than between R and T. Thus, these structures support only a T left and right arrow R left and right arrow R2 allosteric pathway and underscore the physiological importance of the R2 conformation. PMID- 9223275 TI - Total chemical synthesis of enzymatically active human type II secretory phospholipase A2. AB - Human group II secretory phospholipase A2 (sPLA2) is an enzyme found in the alpha granules of platelets and at inflammatory sites. Although its physiological function is unclear, sPLA2 can inhibit blood coagulation reactions independent of its lipolytic action. To study the molecular basis of PLA2 activities, we developed a total chemical synthesis of sPLA2 by chemical ligation of large unprotected peptides. The synthetic segments PLA2-(1-58)-alphaCOSCH2COOH and PLA2 (59-124) were prepared by stepwise solid-phase peptide synthesis and ligated to yield a peptide bond between Gly58 and Cys59. The 124-residue polypeptide product (mass: 13,920 +/- 2 Da) was folded to yield one major product (mass: 13,905 +/- 1 Da), the loss of 15 +/- 3 Da reflecting the formation of seven disulfide bonds. Circular dichroism studies of synthetic sPLA2 showed alpha-helix, beta-structure, and random coil contents consistent with those found in the crystal structure of sPLA2. Synthetic sPLA2 had kcat and Km values identical to those of recombinant sPLA2 for hydrolysis of 1,2-bis(heptanoylthio)-phosphatidylcholine. Synthetic sPLA2, like recombinant sPLA2, inhibited thrombin generation from prothrombinase complex (factors Xa, V, II, Ca2+, and phospholipids). In the absence of phospholipids, both synthetic and recombinant sPLA2 inhibited by 70% prothrombin activation by factors Xa, Va, and Ca2+. Thus, synthetic sPLA2 is a phospholipid independent anticoagulant like recombinant or natural sPLA2. This study demonstrates that chemical synthesis of sPLA2 yields a fully active native-like enzyme and offers a straightforward tool to provide sPLA2 analogs for structure activity studies of anticoagulant, lipolytic, or inflammatory activities. PMID- 9223276 TI - A DnaB intein in Rhodothermus marinus: indication of recent intein homing across remotely related organisms. AB - A dnaB gene encoding a homologue of the Escherichia coli DNA helicase DnaB was cloned and sequenced in the thermophilic eubacterium Rhodothermus marinus, predicting a DnaB protein that harbors an intein. This DnaB intein is 428 amino acid residues long, has several putative intein sequence motifs (including two putative endonuclease motifs), and is capable of protein splicing when produced in E. coli cells. The R. marinus DnaB intein is a close homologue of a DnaB intein in the cyanobacterium Synechocystis sp. strain PCC6803. The two inteins are positioned identically in their respective DnaB proteins. They also share a 54% sequence identity (74% sequence similarity) that is markedly higher than the 37% sequence identity shared by the extein sequences of the two DnaB proteins. Horizontal intein transfer (homing) is therefore invoked to relate these two DnaB inteins. The codon usage of R. marinus DnaB intein coding sequence differs markedly from the codon usages of its flanking extein coding sequences and other genes in the same genome, suggesting more recent acquisition of the DnaB intein in this organism. PMID- 9223277 TI - X-ray structures of a hydrolytic antibody and of complexes elucidate catalytic pathway from substrate binding and transition state stabilization through water attack and product release. AB - The x-ray structures of the unliganded esterase-like catalytic antibody D2.3 and its complexes with a substrate analogue and with one of the reaction products are analyzed. Together with the structure of the phosphonate transition state analogue hapten complex, these crystal structures provide a complete description of the reaction pathway. At alkaline pH, D2.3 acts by preferential stabilization of the negatively charged oxyanion intermediate of the reaction that results from hydroxide attack on the substrate. A tyrosine residue plays a crucial role in catalysis: it activates the ester substrate and, together with an asparagine, it stabilizes the oxyanion intermediate. A canal allows facile diffusion of water molecules to the reaction center that is deeply buried in the structure. Residues bordering this canal provide targets for mutagenesis to introduce a general base in the vicinity of the reaction center. PMID- 9223278 TI - Direct involvement of the ubiquitin-conjugating enzyme Ubc9/Hus5 in the degradation of IkappaBalpha. AB - The NF-kappaB/Rel proteins are sequestered in the cytoplasm in association with IkappaBalpha. In response to external signals, IkappaBalpha is phosphorylated, multi-ubiquitinated, and degraded by proteasomes, thereby releasing NF-kappaB/Rel proteins to migrate to the nucleus. We have cloned a mouse ubiquitin-conjugating enzyme (mE2), which associates with IkappaBalpha. mE2 is homologous to the yeast Ubc9/Hus5 ubiquitin-conjugating enzyme. A transdominant-negative mutant of mE2 had no effect on phosphorylation of IkappaBalpha, but delayed its degradation. Correspondingly, tumor necrosis factor-alpha-inducible NF-kappaB activity was diminished. We propose that mE2 is directly involved in the ubiquitin conjugation of IkappaBalpha, a pivotal step in its degradation pathway. PMID- 9223279 TI - The p53 tumor suppressor targets a novel regulator of G protein signaling. AB - Heterotrimeric G proteins transduce multiple growth-factor-receptor-initiated and intracellular signals that may lead to activation of the mitogen-activated or stress-activated protein kinases. Herein we report on the identification of a novel p53 target gene (A28-RGS14) that is induced in response to genotoxic stress and encodes a novel member of a family of regulators of G protein signaling (RGS) proteins with proposed GTPase-activating protein activity. Overexpression of A28 RGS14p protein inhibits both Gi- and Gq-coupled growth-factor-receptor-mediated activation of the mitogen-activated protein kinase signaling pathway in mammalian cells. Thus, through the induction of A28-RGS14, p53 may regulate cellular sensitivity to growth and/or survival factors acting through G protein-coupled receptor pathways. PMID- 9223280 TI - Human and Saccharomyces cerevisiae dolichol phosphate mannose synthases represent two classes of the enzyme, but both function in Schizosaccharomyces pombe. AB - Dolichol phosphate mannose (Dol-P-Man), formed upon transfer of Man from GDPMan to Dol-P, is a mannosyl donor in pathways leading to N-glycosylation, glycosyl phosphatidylinositol membrane anchoring, and O-mannosylation of protein. Dol-P Man synthase is an essential protein in Saccharomyces cerevisiae. We have cloned cDNAs encoding human and Schizosaccharomyces pombe proteins that resemble S. cerevisiae Dol-P-Man synthase. Disruption of the gene for the S. pombe Dol-P-Man synthase homolog, dpm1(+), is lethal. The known Dol-P-Man synthase sequences can be divided into two classes. One contains the S. cerevisiae, Ustilago maydis, and Trypanosoma brucei enzymes, which have a COOH-terminal hydrophobic domain, and the other contains the human, S. pombe, and Caenorhabditis synthases, which lack a hydrophobic COOH-terminal domain. The two classes of synthase are functionally equivalent, because S. cerevisiae DPM1 and its human counterpart both complement the lethal null mutation in S. pombe dpm1(+). The findings that Dol-P-Man synthase is essential in yeast and that the Ustilago and Trypanosoma synthases are in a different class from the human enzyme raise the possibility that Dol-P Man synthase could be exploited as a target for inhibitors of pathogenic eukaryotic microbes. PMID- 9223281 TI - Steroid receptor induction of gene transcription: a two-step model. AB - Coactivators, such as steroid receptor coactivator 1 (SRC-1A) and CREB (cAMP response element binding protein)-binding protein (CBP), are required for efficient steroid receptor transactivation. Using an in vitro transcription assay, we found that progesterone receptor (PR)-driven transcription is inhibited by a dominant negative PR ligand-binding domain-interacting region of SRC-1A, indicating that SRC-1A is required for actual transcriptional processes. In addition, these coactivators also possess intrinsic histone acetyltransferase (HAT) activity and bind to each other and another HAT, p300/CBP-associated factor. Here we show that the human PR also interacts with p300/CBP-associated factor in vitro. Recruitment of multiple HATs to target promoters suggests an important role for chromatin remodeling in transcriptional activation of genes by steroid receptors. In transient transfection assays, we found that addition of a histone deacetylase inhibitor, trichostatin A, strongly potentiated PR-driven transcription. In contrast, directing histone deacetylase-1 (HD1) to a promoter using the GAL4 DNA binding domain inhibited transcription. Furthermore, PR transactivation was repressed by recruiting HD1 into the PR-DNA complex by fusing HD1 to a PR ligand-binding domain-interacting portion of SRC-1. Collectively, these results suggest that targeted histone acetylation by recruited HAT cofactors and histone deacetylation are important factors affecting PR transactivation. Recruitment of coactivators and HATs by the liganded PR in vivo may result in (i) remodeling of transcriptionally repressed chromatin to facilitate assembly and (ii) enhanced stabilization of the preinitiation complex by the activation functions of coactivators and the liganded PR itself. PMID- 9223282 TI - Alpha-tocopheryl hydroquinone is an efficient multifunctional inhibitor of radical-initiated oxidation of low density lipoprotein lipids. AB - As the oxidation of low density lipoprotein (LDL) lipids may be a key event in atherogenesis, there is interest in antioxidants as potential anti-atherogenic compounds. Here we report that alpha-tocopheryl hydroquinone (alpha-TQH2) strongly inhibited or completely prevented the (per)oxidation of ubiquinol-10 (CoQ10H2), alpha-tocopherol (alpha-TOH), and both surface and core lipids in LDL exposed to either aqueous or lipophilic peroxyl radicals, Cu2+, soybean lipoxygenase, or the transition metal-containing Ham's F-10 medium in the absence or presence of human monocyte-derived macrophages. The antioxidant activity of alpha-TQH2 was superior to that of several other lipophilic hydroquinones, including endogenous CoQ10H2, which is regarded as LDL's first line of antioxidant defence. At least three independent activities contributed to the antioxidant action of alpha-TQH2. First, alpha-TQH2 readily associated with LDL and instantaneously reduced the lipoprotein's ubiquinone-10 to CoQ10H2, thereby maintaining this antioxidant in its active form. Second, alpha-TQH2 directly intercepted aqueous peroxyl radicals, as indicated by the increased rate of its consumption with increasing rates of radical production, independent of LDL's content of CoQ10H2 and alpha-TOH. Third, alpha-TQH2 rapidly quenched alpha tocopheroxyl radical in oxidizing LDL, as demonstrated directly by electron paramagnetic resonance spectroscopy. Similar antioxidant activities were also seen when alpha-TQH2 was added to high-density lipoprotein or the protein-free Intralipid, indicating that the potent antioxidant activity of alpha-TQH2 was neither lipoprotein specific nor dependent on proteins. These results suggest that alpha-TQH2 is a candidate for a therapeutic lipid-soluble antioxidant. As alpha-tocopherylquinone is formed in vivo at sites of oxidative stress, including human atherosclerotic plaque, and biological systems exist that reduce the quinone to the hydroquinone, our results also suggest that alpha-TQH2 could be a previously unrecognized natural antioxidant. PMID- 9223283 TI - The herpes simplex virus 1 protein kinase US3 is required for protection from apoptosis induced by the virus. AB - An earlier report showed that a disabled mutant lacking both copies of the major regulatory gene (alpha4) of herpes simplex virus 1 induced DNA degradation characteristic of apoptosis in infected cells, whereas the wild-type virus protected cells from apoptosis induced by thermal shock. More extensive analyses of the disabled mutant revealed a second mutation which disabled US3, a viral gene encoding a protein kinase known to phosphorylate serine/threonine within a specific arginine-rich consensus sequence. Analyses of cells infected with a viral mutant carrying a wild-type alpha4 gene but from which the US3 gene had been deleted showed that it induced fragmentation of cellular DNA, whereas a recombinant virus in which the deleted sequences of the US3 gene had been restored did not cause the cellular DNA to fragment. These results point to the protein kinase encoded by the US3 gene as the principal viral product required to block apoptosis. PMID- 9223284 TI - The major yeast poly(A)-binding protein is associated with cleavage factor IA and functions in premessenger RNA 3'-end formation. AB - Polyadenylation of premessenger RNAs occurs posttranscriptionally in the nucleus of eukaryotic cells by cleavage of the precursor and polymerization of adenosine residues. In the yeast Saccharomyces cerevisiae, the mature poly(A) tail ranges from 60 to 70 nucleotides. 3'-end processing can be reproduced in vitro with purified factors. The cleavage reaction requires cleavage factors I and II (CF I and CF II), whereas polyadenylation involves CF I, polyadenylation factor I (PFI), and poly(A) polymerase (Pap1p). CF I has recently been separated into two factors, CF IA and CF IB. We have independently purified CF IA and found that five polypeptides cofractionate with the activity. They include Rna14p, Rna15p, Pcf11p, a new protein called Clp1p, and remarkably, the major poly(A)-binding protein Pab1p. Extracts from strains where the PAB1 gene is mutated or deleted are active for cleavage but generate transcripts bearing abnormally long poly(A) tracts. Complementation with recombinant Pab1p not only restores the length of the poly(A) tails to normal, but also triggers a poly(A) shortening activity. In addition, a monoclonal Pab1p antibody prevents the formation of poly(A) tails in extracts or in a reconstituted system. Our data support the notion that Pab1p is involved in the length control of the poly(A) tails of yeast mRNAs and define a new essential function for Pab1p in the formation of mature mRNAs. PMID- 9223285 TI - Leishmania tarentolae contains distinct cytosolic and mitochondrial glutaminyl tRNA synthetase activities. AB - The intracellular distribution of glutaminyl-tRNA synthetases and their role in mitochondrial tRNA import were evaluated in the ancient eukaryote Leishmania tarentolae. The following results were obtained: (i) Glutaminyl-tRNA synthetase was detected in leishmanial mitochondria. This was unexpected because it has been postulated that, in organelles, Gln-tRNAGln is not formed by direct acylation of tRNAGln but by enzymatic transamidation of misacylated Glu-tRNAGln. (ii) Whereas the cytosolic extract is able to charge cytosolic and mitochondrial tRNAsGln, the mitochondrial matrix extract does not aminoacylate the cytosol-specific tRNAGln. This indicates that mitochondrial and cytosolic glutaminyl-tRNA synthetases are distinct. (iii) Seven of the 11 nucleotides that differ between the cytosolic and the mitochondrial tRNAGln are sufficient to convert the cytosol-specific tRNAGln into an optimal substrate for the mitochondrial enzyme. These nucleotides are arranged in three groups consisting of the nucleotides flanking the anticodon stem, the 5' nucleotide of the anticodon, and four nucleotides within the acceptor stem. And (iv), it was shown that the identity elements for recognition by the mitochondrial glutaminyl-tRNA synthetase do not overlap with a previously identified sequence segment required for mitochondrial import of the tRNAGln. PMID- 9223286 TI - Activation of the orphan receptor RIP14 by retinoids. AB - Retinoids are crucial regulators of a wide variety of processes in both developing and adult animals. These effects are thought to be mediated by the retinoic acid (RA) receptors and the retinoid X receptors (RXRs). We have identified an additional retinoid-activated receptor that is neither a retinoic acid receptors nor an RXR. RXR-interacting protein 14 (RIP14), a recently described orphan member of the nuclear receptor superfamily, can be activated by either all-trans-RA (tRA) or the synthetic retinoid TTNPB [[E]-4-[2-(5, 6, 7, 8 tetrahydro-5, 5, 8, 8-tetramethyl-2-naphthalenyl)propen-1-yl]benzoic acid].RIP14 binds to DNA as a heterodimer with RXR. In the presence of either tRA or TTNPB, the addition of 9-cis-RA or the RXR-specific agonist LG1069 [4-[1-(3, 5, 5, 8, 8 pentamethyl-5, 6, 7, 8-tertrahydro-2-naphthyl)ethenyl]benzoic acid] results in additional activation. Mutations of the ligand-dependent transcriptional activation functions indicate that TTNPB activates the RIP14 component of the RIP14-RXR heterodimer, that 9-cis-RA and LG1069 activate RXR, and that tRA activates via both RIP14 and RXR. Despite the very effective activation of RIP14 by tRA or TTNPB, relatively high concentrations of these compounds are required, and no evidence for direct binding of either compound was obtained using several approaches. These results suggest that RIP14 is the receptor for an as-yet unidentified retinoid metabolite. PMID- 9223287 TI - The Xenopus proglucagon gene encodes novel GLP-1-like peptides with insulinotropic properties. AB - The proglucagon gene encodes several hormones that have key roles in the regulation of metabolism. In particular, glucagon-like peptide (GLP-1), a potent stimulus of insulin secretion, is being developed as a therapy for the treatment of non-insulin-dependent diabetes mellitus. To define structural moieties of the molecule that convey its insulinotropic activity, we have cloned and characterized the proglucagon gene from the amphibian, Xenopus laevis. Unexpectedly, these cDNAs were found to encode three unique glucagon-like-1 peptides, termed xenGLP-1A, xenGLP-1B, and xenGLP-1C in addition to the typical proglucagon-derived hormones glucagon and GLP-2. xenGLP-1A, -1B, and -1C were synthesized and tested for their ability to bind and activate the human GLP-1 receptor (hGLP-1R), and to stimulate insulin release from rat pancreas. All three Xenopus GLP-1-like peptides bind effectively to the hGLP-1R and stimulate cAMP production. Surprisingly, xenGLP-1B(1-30) demonstrated higher affinity for the hGLP-1R than hGLP-1 (IC50 of 1.1 +/- 0.4 nM vs. 4.4 +/- 1.0 nM, respectively, P < 0.02) and was equipotent to hGLP-1 in stimulating cAMP production (EC50 of 0.17 +/- 0.02 nM vs. 0.6 +/- 0. 2 nM, respectively, P > 0.05). Further studies demonstrated that hGLP-1, xenGLP-1A, -1B, and -1C stimulate comparable insulin release from the pancreas. These results demonstrate that despite an average of nine amino acid differences between the predicted Xenopus GLPs and hGLP-1, all act as hGLP-1R agonists. PMID- 9223289 TI - Detection of single-molecule interactions using correlated thermal diffusion. AB - Observation of discrete, single-molecule binding events allows one to bypass assumptions required to infer single-molecule properties from studies of ensembles of molecules. Optically trapped beads and glass microneedles have been applied to detect single-molecule binding events, but it remains difficult to identify signs of binding events given the large displacements induced by thermal forces. Here, we exploit thermal diffusion by using correlation between motion of optically trapped beads attached to both ends of a single actin filament to track binding events of individual myosin molecules. We use correlated diffusion to measure the stiffness of a single myosin molecule and estimate its thermal fluctuation in a poststroke state as comparable in amplitude to the measured stroke distance. The use of correlated diffusion to measure kinetics of single molecule interactions and the stiffness of the interacting moieties should be applicable to any pair of interacting molecules, and not limited to biological motors. PMID- 9223288 TI - Targeted inactivation of alphai2 or alphai3 disrupts activation of the cardiac muscarinic K+ channel, IK+Ach, in intact cells. AB - Cardiac muscarinic receptors activate an inwardly rectifying K+ channel, IK+Ach, via pertussis toxin (PT)-sensitive heterotrimeric G proteins (in heart Gi2, Gi3, or Go). We have used embryonic stem cell (ES cell)-derived cardiocytes with targeted inactivations of specific PT-sensitive alpha subunits to determine which G proteins are required for receptor-mediated regulation of IK+Ach in intact cells. The muscarinic agonist carbachol increased IK+Ach activity in ES cell derived cardiocytes from wild-type cells, in cells lacking alphao, and in cells lacking the PT-insensitive G protein alphaq. In cells with targeted inactivation of alphai2 or alphai3, channel activation by both carbachol and adenosine was blocked. Carbachol-induced channel activation was restored in the alphai2- and alphai3-null cells by reexpressing the previously targeted gene and guanosine 5' [gamma-thio] triphosphate was able to fully activate IK+Ach in excised membranes patches from these mutants. In contrast, negative chronotropic responses to both carbachol and adenosine were preserved in cells lacking alphai2 or alphai3. Our results show that expression of two specific PT-sensitive alpha subunits (alphai2 and alphai3 but not alphao) is required for normal agonist-dependent activation of IK+Ach and suggest that both alphai2- and alphai3-containing heterotrimeric G proteins may be involved in the signaling process. Also the generation of negative chronotropic responses to muscarinic or adenosine receptor agonists do not require activation of IK+Ach or the expression of alphai2 or alphai3. PMID- 9223290 TI - Nonexponential kinetics of a single tRNAPhe molecule under physiological conditions. AB - The fluorescence decay functions of individual, specifically labeled tRNAPhe molecules exhibit nonexponential character as a result of conformational dynamics occurring during the measurement on a single molecule. tRNAPhe conformational states that interchange much more slowly are evidenced by the distribution of lifetimes observed for many individual molecules. A structural model for the nonexponential decay indicates that the tRNAPhe-probe adduct fluctuates between two states, one of which provides conditions that quench the probe fluorescence. PMID- 9223291 TI - Microsecond atomic force sensing of protein conformational dynamics: implications for the primary light-induced events in bacteriorhodopsin. AB - In this paper a new atomic force sensing technique is presented for dynamically probing conformational changes in proteins. The method is applied to the light induced changes in the membrane-bound proton pump bacteriorhodopsin (bR). The microsecond time-resolution of the method, as presently implemented, covers many of the intermediates of the bR photocycle which is well characterized by spectroscopical methods. In addition to the native pigment, we have studied bR proteins substituted with chemically modified retinal chromophores. These synthetic chromophores were designed to restrict their ability to isomerize, while maintaining the basic characteristic of a large light-induced charge redistribution in the vertically excited Franck-Condon state. An analysis of the atomic force sensing signals lead us to conclude that protein conformational changes in bR can be initiated as a result of a light-triggered redistribution of electronic charge in the retinal chromophore, even when isomerization cannot take place. Although the coupling mechanism of such changes to the light-induced proton pump is still not established, our data question the current working hypothesis which attributes all primary events in retinal proteins to an initial trans<==>cis isomerization. PMID- 9223292 TI - Kinetic theory of fibrillogenesis of amyloid beta-protein. AB - Prior quasielastic light scattering (QLS) studies of fibrillogenesis of synthetic amyloid beta-protein (Abeta)-(1-40) at low pH have suggested a kinetic model in which: (i) fibrillogenesis requires a nucleation step; (ii) nuclei are produced by Abeta micelles in addition to seeds initially present; and (iii) fibril elongation occurs by irreversible binding of Abeta monomers to the fibril ends. Here we present the full mathematical formulation of this model. We describe the temporal evolution of the concentrations of Abeta monomers and micelles as well as the concentration and size distribution of fibrils. This formulation enables deduction of the fundamental parameters of the model-e.g., the nucleation and elongation rate constants kn and ke-from the time dependency of the apparent diffusion coefficient measured by QLS. The theory accurately represents the experimental observations for Abeta concentrations both below and above c*, the critical concentration for Abeta micelle formation. We suggest that the method of QLS in combination with this theory can serve as a powerful tool for understanding the molecular factors that control Abeta plaque formation. PMID- 9223293 TI - Nuclear localization of basonuclin in human keratinocytes and the role of phosphorylation. AB - Basonuclin is a zinc-finger protein found in basal cells of the epidermis. In human keratinocyte cultures, basonuclin is susceptible to serine-phosphorylation and the addition of the phosphatase inhibitor, okadaic acid, promotes accumulation of basonuclin in the cytoplasm. The region of basonuclin containing the nuclear localization signal of basonuclin is necessary for nuclear localization of the protein and Ser-541, located immediately C-terminal to the nuclear localization signal, is the principal phosphorylation site in vitro. A nearly complete basonuclin transiently expressed in cultured keratinocytes localizes predominantly in the nucleus, but substitution of aspartic acid for Ser 541 promotes cytoplasmic localization. The same substitution of Ser-537 has a similar but weaker effect. Substitution of both serine residues by alanine leads to nuclear localization. These results show that nuclear localization of basonuclin depends on serine dephosphorylation, primarily of Ser-541. Different subcellular locations of basonuclin in different keratinocyte subtypes are therefore most likely to be controlled by the state of phosphorylation of Ser 541. PMID- 9223294 TI - Tumor induction by a transformation-defective polyoma virus mutant blocked in signaling through Shc. AB - Transformation of cells in culture by polyoma virus requires integration of signals downstream of middle T-Shc and middle T-phosphatidylinositol 3-kinase interactions, but the same is not true for induction of tumors in the mouse. Thus, a middle T mutant defective in transformation and blocked in binding Shc is able to induce a broad spectrum of tumors after inoculation into newborn mice. The "tumor profile" induced by the mutant shows enhancement of tumors at some sites and reductions at others but otherwise resembles that induced by the wild type virus. A nontransforming double-mutant blocked in binding phosphatidylinositol 3-kinase as well as Shc is severely affected but still induces some tumors. These results show that pathways that must cooperate to induce full transformation of cells in vitro can act independently and are to a large extent redundant in tumor induction. PMID- 9223295 TI - Transmembrane/cytoplasmic domain-mediated membrane type 1-matrix metalloprotease docking to invadopodia is required for cell invasion. AB - The invasion of human malignant melanoma cells into the extracellular matrix (ECM) involves the accumulation of proteases at sites of ECM degradation where activation of matrix metalloproteases (MMP) occurs. Here, we show that when membrane type 1 MMP (MT-MMP) was overexpressed in RPMI7951 human melanoma cells, the cells made contact with the ECM, activated soluble and ECM-bound MMP-2, and degraded and invaded the ECM. Further experiments demonstrated the importance of localization of the MT-MMP to invadopodia. Overexpression of MT-MMP without invadopodial localization caused activation of soluble MMP-2, but did not facilitate ECM degradation or cell invasiveness. Up-regulation of endogenous MT MMP with concanavalin A caused activation of MMP-2. However, concanavalin A treatment prevented invadopodial localization of MT-MMP and ECM degradation. Neither a truncated MT-MMP mutant lacking transmembrane (TM) and cytoplasmic domains (DeltaTMMT-MMP), nor a chimeric MT-MMP containing the interleukin 2 receptor alpha chain (IL-2R) TM and cytoplasmic domains (DeltaTMMT-MMP/TMIL-2R) were localized to invadopodia or exhibited ECM degradation. Furthermore, a chimera of the TM/cytoplasmic domain of MT-MMP (TMMT-MMP) with tissue inhibitor of MMP 1 (TIMP-1/TMMT-MMP) directed the TIMP-1 molecule to invadopodia. Thus, the MT-MMP TM/cytoplasmic domain mediates the spatial organization of MT-MMP into invadopodia and subsequent degradation of the ECM. PMID- 9223296 TI - The Schizosaccharomyces pombe spindle checkpoint protein mad2p blocks anaphase and genetically interacts with the anaphase-promoting complex. AB - The spindle checkpoint monitors mitotic spindle integrity and the attachment of kinetochores to the spindle. Upon sensing a defect the checkpoint blocks cell cycle progression and thereby prevents chromosome missegregation. Previous studies in budding yeast show that the activated spindle checkpoint inhibits the onset of anaphase by an unknown mechanism. One possible target of the spindle checkpoint is anaphase promoting complex (APC), which controls all postmetaphase events that are blocked by spindle checkpoint activation. We have isolated mad2, a spindle checkpoint component in fission yeast, and shown that mad2 overexpression activates the checkpoint and causes a cell cycle arrest at the metaphase-to-anaphase transition. In addition to the observation that mad2 induced arrest can be partially relieved by mitosis-promoting factor inactivation, we present genetic evidence consistent with the hypothesis that the spindle checkpoint imposes a cell cycle arrest by inhibiting APC-dependent proteolysis. PMID- 9223297 TI - Roles of phospholipase C beta2 in chemoattractant-elicited responses. AB - The physiological roles of phospholipase C (PLC) beta2 in hematopoiesis, leukocyte function, and host defense against infection were investigated using a mouse line that lacks PLC beta2. PLC beta2 deficiency did not affect hematopoiesis, but it blocked chemoattractant-induced Ca2+ release, superoxide production, and MAC-1 up-regulation in neutrophils. In view of these effects, it was surprising that the absence of PLC beta2 enhanced chemotaxis of different leukocyte populations and sensitized the in vivo response of the PLC beta2 deficient mice to bacteria, viruses, and immune complexes. These data raise questions about the roles that PLC beta2 may play in signal transduction induced by chemoattractants in leukocytes. PMID- 9223298 TI - An upstream, DNase I hypersensitive region of the hematopoietic-expressed transcription factor GATA-1 gene confers developmental specificity in transgenic mice. AB - The transcription factor GATA-1, which is expressed in several hematopoietic lineages and multipotential progenitors, is required for the development of red blood cells and platelets. To identify control elements of the mouse GATA-1 gene, we analyzed DNase I hypersensitivity of the locus in erythroid chromatin and the expression of GATA-1/Escherichia coli beta-galactosidase (lacZ) transgenes in mice. Transgenes with 2.7 kb of promoter sequences are expressed infrequently and only within adult (definitive) erythroid cells. We show that inclusion of an upstream hypersensitive site (HS I) markedly enhances the frequency of expressing transgenic lines and activates expression in primitive erythroid cells. This pattern recapitulates the proper pattern of GATA-1 expression during development. By breeding a GATA-1/lacZ transgene into a GATA-1(-) background, we also have shown that the activation or maintenance of GATA-1 expression does not require the presence of GATA-1 itself, thereby excluding simple models of positive autoregulation. The transgene cassette reported here should be useful in directing expression of foreign sequences at the onset of hematopoiesis in the embryo and may assist in the identification of upstream regulators of the GATA-1 gene. PMID- 9223300 TI - Phylogeny of the genus Pistacia as determined from analysis of the chloroplast genome. AB - Classification within the genus Pistacia has been based on leaf morphology and geographical distribution. Molecular genetic tools (PCR amplification followed by restriction analysis of a 3.2-kb region of variable chloroplast DNA, and restriction fragment length polymorphism analysis of the Pistacia cpDNA with tobacco chloroplast DNA probes) provided a new set of variables to study the phylogenetic relationships of 10 Pistacia species. Both parsimony and cluster analyses were used to divide the genus into two major groups. P. vera was determined to be the least derived species. P. weinmannifolia, an Asian species, is most closely related to P. texana and P. mexicana, New World species. These three species share a common origin, suggesting that a common ancestor of P. texana and P. mexicana originated in Asia. P. integerrima and P. chinensis were shown to be distinct whereas the pairs of species were monophyletic within each of two tertiary groups, P. vera:P. khinjuk and P. mexicana:P. texana. An evolutionary trend from large to small nuts and leaves with few, large leaflets to many, small leaflets was supported. The genus Pistacia was shown to have a low chloroplast DNA mutation rate: 0.05-0.16 times that expected of annual plants. PMID- 9223301 TI - An upper dentition of Aframonius dieides (Primates) from the Fayum, Egyptian Eocene. AB - The first known upper dentitions--an adult and subadult--of the cercamoniine adapiform Aframonius dieides are described. Comparisons show that A. dieides has an upper molar morphology resembling that of other cercamoniine adapids but the species lacks some of their typical specializations. The new dental material confirms that Aframonius stands closer to Mahgarita from west Texas and Cercamonius from Europe than it does to Schizarodon and Omanodon from Oman-all of which have been ranked as cercamoniines. Affinities of the latter two genera probably lie with the Anchomomys group. The presence of a cercamoniine adapid in the Eocene of Egypt supports the view that early African anthropoideans evolved not in isolation, but concomitantly with a contemporary Eocene prosimian radiation. PMID- 9223299 TI - An important developmental role for oligosaccharides during early embryogenesis of cyprinid fish. AB - Derivatives of chitin oligosaccharides have been shown to play a role in plant organogenesis at nanomolar concentrations. Here we present data which indicate that chitin oligosaccharides are important for embryogenesis in vertebrates. We characterize chitin oligosaccharides synthesized in vitro by zebrafish and carp embryos in the late gastrulation stage by incorporation of radiolabeled N-acetyl D-[U14C]glucosamine and by HPLC in combination with enzymatic conversion using the Bradyrhizobium NodZ alpha-1, 6-fucosyltransferase and chitinases. A rapid and sensitive bioassay for chitin oligosaccharides was also used employing suspension cultured plant cells of Catharanthus roseus. We show that chitin oligosaccharide synthase activity is apparent only during late gastrulation and can be inhibited by antiserum raised against the Xenopus DG42 protein. The DG42 protein, a glycosyltransferase, is transiently expressed between midblastula and neurulation in Xenopus and zebrafish embryogenesis. Microinjection of the DG42 antiserum or the Bradyrhizobium NodZ enzyme in fertilized eggs of zebrafish led to severe defects in trunk and tail development. PMID- 9223302 TI - Functional and nonfunctional mutations distinguished by random recombination of homologous genes. AB - We describe a convenient method for distinguishing functional from nonfunctional or deleterious mutations in homologous genes. High fidelity in vitro gene recombination ("DNA shuffling") coupled with sequence analysis of a small sampling of the shuffled library exhibiting the evolved behavior allows identification of those mutations responsible for the behavior in a background of neutral and deleterious mutations. Functional mutations are expected to occur in 100% of the sequenced screened sample; neutral mutations are found in 50% on average, and deleterious mutations do not appear at all. When used to analyze 10 mutations in a laboratory-evolved gene encoding a thermostable subtilisin E, this method rapidly identified the two responsible for the observed protease thermostability; the remaining eight were neutral with respect to thermostability, within the precision of the screening assay. A similar approach, coupled with selection for growth and survival of the host organism, could be used to distinguish adaptive from neutral mutations. PMID- 9223303 TI - Cleavage patterns and the topology of the metazoan tree of life. AB - Several major alliances of metazoan phyla have been identified by small subunit rRNA sequence comparisons. It is possible to arrange the phyla to produce a parsimonious distribution of cleavage types, requiring only one change from a radial ancestral condition to spiral cleavage and one other to "idiosyncratic" cleavage; this arrangement is consistent with most of the recent molecular phylogenies. The cleavage shifts are correlated with changes in many of the features that once were used to distinguish Protostomia and Deuterostomia. It is hypothesized that changes in cleavage direction are causally associated with changes in blastomere fates and thus that cleavage type correlates with such features as the identity of mesoderm founder cells, which in turn can constrain the mode of origination of the eucelom. Cleavage changes may also affect the timing of cell fate specification. In a tree that emphasizes cleavage parsimony, radial cleavage, regulative development, and enterocely are ancestral within the Bilateria, and spiral or idiosyncratic cleavages, mosaic development, and schizocely are associated with a change in cleavage direction. Deuterostomy is presumably ancestral and is correlated with radial cleavage for this reason, rather than mechanistically. PMID- 9223304 TI - Molecular evidence that echiurans and pogonophorans are derived annelids. AB - The Annelida, which includes the polychaetes and the clitellates, has long held the taxonomic rank of phylum. The unsegmented, mud-dwelling echiuran spoon worms and the gutless, deep-sea pogonophoran tube worms (including vestimentiferans) share several embryological and morphological features with annelids, but each group also has been considered as a separate metazoan phylum based on the unique characters each group displays. Phylogenetic analyses of DNA sequences from the nuclear gene elongation factor-1alpha place echiurans and pogonophorans within the Annelida. This result, indicating the derived loss of segmentation in echiurans, has profound implications for our understanding of the evolution of metazoan body plans and challenges the traditional view of the phylum-level diversity and evolutionary relationships of protostome worms. PMID- 9223305 TI - Cloning and characterization of hOGG1, a human homolog of the OGG1 gene of Saccharomyces cerevisiae. AB - The OGG1 gene of Saccharomyces cerevisiae encodes a DNA glycosylase activity that is a functional analog of the Fpg protein from Escherichia coli and excises 7,8 dihydro-8-oxoguanine (8-oxoG) from damaged DNA. The repair of this ubiquitous kind of oxidative damage is essential to prevent mutations both in bacteria and in yeast. A human cDNA clone carrying an ORF displaying homology to the yeast protein was identified. The predicted protein has 345 amino acids and a molecular mass of 39 kDa. This protein shares a 38% sequence identity with the yeast Ogg1 protein, adding this novel human gene product to the growing family of enzymes that the repair of oxidatively damaged bases and are related to the E. coli endonuclease III. Northern blot analysis indicates that this gene, localized to chromosome 3p25, is ubiquitously expressed in human tissues. The cloned coding sequence was expressed in an E. coli strain that carried a disrupted fpg gene, the bacterial functional analog of OGG1. Cell-free extracts from these cultures displayed a specific lyase activity on duplex DNA that carried an 8-oxoG/C base pair. The products of the reaction are consistent with an enzymatic activity like the one displayed by the yeast Ogg1. Analysis of the substrate specificity reveals a very strong preference for DNA fragments harboring 8-oxoG/C base pairs. The pattern of specificity correlates well with the one found for the yeast enzyme. Moreover, when the human coding sequence was expressed in a yeast strain mutant in OGG1 it was able to complement the spontaneous mutator phenotype. These results make this novel gene (hOGG1) a strong candidate for the human homolog of the yeast OGG1 and suggest an important role of its product in the protection of the genome from the mutagenic effects of the oxidatively damaged purines. PMID- 9223306 TI - Molecular cloning and functional expression of a human cDNA encoding the antimutator enzyme 8-hydroxyguanine-DNA glycosylase. AB - The major mutagenic base lesion in DNA caused by exposure to reactive oxygen species is 8-hydroxyguanine (8-oxo-7, 8-dihydroguanine). In bacteria and Saccharomyces cerevisiae, this damaged base is excised by a DNA glycosylase with an associated lyase activity for chain cleavage. We have cloned, sequenced, and expressed a human cDNA with partial sequence homology to the relevant yeast gene. The encoded 47-kDa human enzyme releases free 8-hydroxyguanine from oxidized DNA and introduces a chain break in a double-stranded oligonucleotide specifically at an 8-hydroxyguanine residue base paired with cytosine. Expression of the human protein in a DNA repair-deficient E. coli mutM mutY strain partly suppresses its spontaneous mutator phenotype. The gene encoding the human enzyme maps to chromosome 3p25. These results show that human cells have an enzyme that can initiate base excision repair at mutagenic DNA lesions caused by active oxygen. PMID- 9223309 TI - The one ancestor per generation rule and three other rules of mitochondrial inheritance. AB - In mammals, at least, a species-specific mechanism exists that eliminates sperm derived mitochondrial DNA from a fertilized egg. The result is the "one female ancestor per generation" rule and three other rules of mitochondrial inheritance. The second, third, and fourth rules are as follows. (ii) Sublineages of a given mitochondrial line can be generated only during the parallel descents from ancestral sisters. (iii) In a static population in which the production of one female progeny per mated pair per generation has been a rule, several ancient mitochondrial lineages harking back to the female founders of the speciation may persist side by side. (iv) Two or more individuals not related to each other in the recent past may share the identical or nearly identical mitochondrial genome derived from the common female ancestor or ancestral sisters of many generations ago. PMID- 9223308 TI - Transitions in the coupling of transcription and nucleotide excision repair within RNA polymerase II-transcribed genes of Saccharomyces cerevisiae. AB - The molecular mechanism of transcription-coupled nucleotide excision repair in eukaryotes is poorly understood. The identification of the dual role of basal transcription factor TFIIH in DNA repair and transcription provided a plausible link between both processes. However, TFIIH is not part of the elongating transcription complex, suggesting that additional components are required to recruit TFIIH when RNA polymerase II (RNAPII) stalls at the site of DNA damage. Previously, we have shown that the yeast Rad26 protein is involved in transcription-coupled DNA repair. This paper describes the differential contribution of the Rad26 protein to efficient removal of UV-induced cyclobutane pyrimidine dimers (CPDs) from transcribed DNA. Two distinct regions within the transcribed strand of RNAPII-transcribed genes are identified that differ in their requirement for the RAD26 gene product. Using high-resolution repair analysis, we determined the in vivo repair kinetics of cyclobutane pyrimidine dimers positioned around the transcription initiation site of RNAPII-transcribed genes RPB2 and URA3. Although transcription-coupled repair is severely reduced in rad26 mutants, lesions positioned in a small region immediately downstream of transcription initiation are efficiently removed in the absence of Rad26. The observed transition in repair characteristics is abrupt and in excellent agreement with the region where TFIIH dissociates from RNAPII in vitro, strongly suggesting an inverse correlation between TFIIH association and Rad26 requirement. These data suggest that a transcription repair coupling factor (Rad26/CSB) is required for efficient repair only during the elongating stages of RNAPII transcription. PMID- 9223307 TI - Isolation of full-length ATM cDNA and correction of the ataxia-telangiectasia cellular phenotype. AB - A gene mutated in the human genetic disorder ataxia-telangiectasia (A-T), ATM, was recently identified by positional cloning. ATM is a member of the phosphatidylinositol-3-kinase superfamily, some of which are protein kinases and appear to have important roles in cell cycle control and radiation signal transduction. We describe herein, to our knowledge, for the first time, the cloning of a full-length cDNA for ATM and correction of multiple aspects of the radio-sensitive phenotype of A-T cells by transfection with this cDNA. Overexpression of ATM cDNA in A-T cells enhanced the survival of these cells in response to radiation exposure, decreased radiation-induced chromosome aberrations, reduced radio-resistant DNA synthesis, and partially corrected defective cell cycle checkpoints and induction of stress-activated protein kinase. This correction of the defects in A-T cells provides further evidence of the multiplicity of effector functions of the ATM protein and suggests possible approaches to gene therapy. PMID- 9223310 TI - Synergistic and promoter-selective activation of transcription by recruitment of transcription factors TFIID and TFIIB. AB - Eukaryotic transcriptional activators may function by stimulating formation of RNA polymerase II preinitiation complexes at the core promoter of genes. In this case, their mode of action will intrinsically depend on how these complexes assemble on promoters in living cells, an issue that remains largely unexplored. Here we show that in yeast the basal transcription machinery is brought to the promoter in the form of at least two subcomplexes, TFIID and a complex comprising TFIIB and other essential components. Individual recruitment of either complex by artificial contact with a transcriptionally inactive, sequence-specific DNA binding protein suffices to trigger transcriptional activation from a wild-type core promoter bearing the appropriate binding site. In contrast, activation from a promoter containing a weakened TATA element is only observed upon recruitment of TFIID. Tethering TFIIB on that promoter remains without effect, but the simultaneous recruitment of both components leads to strong synergistic activation. These findings suggest a simple mechanism whereby two activators that contact distinct subcomplexes of the basal machinery may stimulate transcription synergistically and differentially depending on the nature of the promoter. PMID- 9223311 TI - Retention of replication fidelity by a DNA polymerase functioning in a distantly related environment. AB - The primary structures of the replicative DNA polymerases (gp43s) of bacteriophage T4 and its distant phylogenetic relative RB69 are diverged, retaining only 61% identity and 74% similarity. Nevertheless, RB69 gp43 substitutes effectively for T4 gp43 in T4 DNA replication in vivo. We show here that RB69 gp43 replicates T4 genomes in vivo with a fidelity similar to that achieved by T4 gp43. Furthermore, replication by RB69 gp43 in the distantly related environment does not enhance the mutator activities of mutations in T4 genes that encode other components of the multienzyme DNA replicase. We also show that the fidelities of RB69 gp43 and T4 gp43 are both high in vitro and that they are similarly and sharply reduced in vivo by mutations that eliminate the 3' exonucleolytic proofreading function. We conclude that gp43 interactions with the other replication proteins are probably nonessential for polymerase fidelity. PMID- 9223312 TI - Fusion genes resulting from alternative chromosomal translocations are overexpressed by gene-specific mechanisms in alveolar rhabdomyosarcoma. AB - Chromosomal translocations identified in hematopoietic and solid tumors result in deregulated expression of protooncogenes or creation of chimeric proteins with tumorigenic potential. In the pediatric solid tumor alveolar rhabdomyosarcoma, a consistent t(2;13)(q35;q14) or variant t(1;13)(p36;q14) translocation generates PAX3-FKHR or PAX7-FKHR fusion proteins, respectively. In this report, we demonstrate that in addition to functional alterations these translocations are associated with fusion product overexpression. Furthermore, PAX3-FKHR and PAX7 FKHR overexpression occurs by distinct mechanisms. Transcription of PAX3-FKHR is increased relative to wild-type PAX3 by a copy number-independent process. In contrast, PAX7-FKHR overexpression results from fusion gene amplification. Thus, gene-specific mechanisms were selected to overexpress PAX3-FKHR and PAX7-FKHR in alveolar rhabdomyosarcoma, presumably due to differences in regulation between the wild-type loci. We postulate that these overexpression mechanisms ensure a critical level of gene product for the oncogenic effects of these fusions. PMID- 9223314 TI - The hotspot conversion paradox and the evolution of meiotic recombination. AB - Studies of meiotic recombination have revealed an evolutionary paradox. Molecular and genetic analysis has shown that crossing over initiates at specific sites called hotspots, by a recombinational-repair mechanism in which the initiating hotspot is replaced by a copy of its homolog. We have used computer simulations of large populations to show that this mechanism causes active hotspot alleles to be rapidly replaced by inactive alleles, which arise by rare mutation and increase by recombination-associated conversion. Additional simulations solidified the paradox by showing that the known benefits of recombination appear inadequate to maintain its mechanism. Neither the benefits of accurate segregation nor those of recombining flanking genes were sufficient to preserve active alleles in the face of conversion. A partial resolution to this paradox was obtained by introducing into the model an additional, nonmeiotic function for the sites that initiate recombination, consistent with the observed association of hotspots with functional sites in chromatin. Provided selection for this function was sufficiently strong, active hotspots were able to persist in spite of frequent conversion to inactive alleles. However, this explanation is unsatisfactory for two reasons. First, it is unlikely to apply to obligately sexual species, because observed crossover frequencies imply maintenance of many hotspots per genome, and the viability selection needed to preserve these would drive the species to extinction. Second, it fails to explain why such a genetically costly mechanism of recombination has been maintained over evolutionary time. Thus the paradox persists and is likely to be resolved only by significant changes to the commonly accepted mechanism of crossing over. PMID- 9223313 TI - High genetic instability of heterochromatin after transposition of the LINE-like I factor in Drosophila melanogaster. AB - In the present work, we have asked whether a group of 13 essential genes mapping to the heterochromatin of Drosophila melanogaster chromosome 2 are mutable following transposition of the I factor during I-R hybrid dysgenesis. We found that the frequency of lethal events mapping to chromosome 2 heterochromatin is surprisingly high, despite the low density of genetic functions identified in this region compared with euchromatin. Cytogenetic and molecular analyses indicated that the recovered mutations correspond either to insertions or to rearrangements. Moreover, chromosomes bearing specific heterochromatic lethal mutations were generated by recombination in the heterochromatin. Together, these data indicate that I factors transpose with high frequency into pericentric regions of chromosome 2 and may play a role in the evolution of constitutive heterochromatin. PMID- 9223315 TI - Direct delivery of exogenous MHC class I molecule-binding oligopeptides to the endoplasmic reticulum of viable cells. AB - After brief incubation of cells with fluorescein-conjugated peptides that bind major histocompatibility complex (MHC) class I molecules, peptides were detected within the endoplasmic reticulum (ER) by microscopy or by binding to radiolabeled class I molecules. ER delivery of a nonfluorescent peptide was demonstrated using a mAb highly specific for the peptide-class I molecule complex. ER localization of peptides: (i) required expression of appropriate class I molecules in the ER but not on the cell surface, (ii) was diminished by expression of TAP, the MHC encoded cytosol to ER peptide transporter, and (iii) was blocked by pinocytosis inhibitors but not by brefeldin A. These findings demonstrate the existence of a pathway, likely vesicular in nature, that conveys small extracellular substances to the ER without traversing the Golgi complex or the cytosol. This pathway contributes to the loading of exogenous peptides to MHC class I molecules, but its evolutionary significance may lie in other cellular processes, such as maintaining ER homeostasis or signaling by extracellular substances. PMID- 9223316 TI - A single peripheral CD8+ T cell can give rise to progeny expressing type 1 and/or type 2 cytokine genes and can retain its multipotentiality through many cell divisions. AB - The lineage relationships between murine CD8(+) T cells with different cytokine profiles were investigated by paired-daughter analysis in the presence and absence of the type 2 cytokine-inducing stimulus, interleukin 4 (IL-4). Single CD8(+) CD44(low) lymph node T cells were activated to divide at high frequency with IL-2 and immobilized antibodies to CD3, CD8, and LFA-1. When these parent cells were subcloned by transferring their daughter or granddaughter cells into secondary cultures with or without IL-4, the subclones expressed diverse combinations of the mRNAs for the type 1 cytokines, interferon gamma (IFN-gamma), and IL-2, and the type 2 cytokines, IL-4, IL-5, IL-6, and IL-10. Frequencies of subclones that expressed IL-4, IL-6, and, to a lesser extent, IL-2, IL-5, and IL 10 were higher among those grown with IL-4, but a significant proportion of those grown without exogenous IL-4 also expressed one or more type 2 cytokines. Subclones within 89% of families displayed different cytokine profiles, indicating that their parent cells were multipotential for this function. Because 98% of parent cells yielded subclones that produced type 1 cytokines and 77% yielded type 2 cytokine producers, we conclude that type 1 and type 2 cytokine producing CD8(+) T cells can be derived from a common precursor. Similar analyses performed by subcloning after >/=7 or >/=13 cell divisions without IL-4 showed that many CD8(+) T cells retained the potential to shift toward a type 2 cytokine profile in response to IL-4, even after prolonged expansion under conditions that favored type 1 cytokine expression. CD8(+) T cells that express type 1 and/or type 2 cytokines therefore are derived from the same peripheral T cell lineage whose multipotentiality can persist through many cell divisions. PMID- 9223317 TI - Ku70-deficient embryonic stem cells have increased ionizing radiosensitivity, defective DNA end-binding activity, and inability to support V(D)J recombination. AB - V(D)J recombination requires both lymphoid-specific and generally expressed enzymatic activities. All three known generally expressed activities involved in V(D)J recombination are also involved in DNA double-strand break repair (DSBR). Two of these are components of the DNA-dependent protein kinase (DNA-PK) and include Ku80 and DNA-PK catalytic subunit (DNA-PKcs); the third, XRCC4, is a protein of unknown function. The Ku70 protein is an additional component of DNA PK; Ku70 forms a heterodimer with Ku80 to generate the DNA end-binding component of the enzyme. To test putative functions for Ku70, we have used gene-targeted mutation to generate a murine embryonic stem cell line which lacks Ku70 expression. We find that the Ku70(-/-) cells produce no detectable Ku70 and very little Ku80, suggesting a direct interrelationship between their levels. Correspondingly, these cells lack the nonspecific DNA end-binding activity associated with Ku. Significantly, the Ku70(-/-) embryonic stem cells have markedly increased sensitivity to gamma-irradiation relative to Ku70(+/-) or wild type embryonic stem cells. Furthermore, the Ku70(-/-) cells lack the ability to effectively rejoin signal and coding ends liberated in transiently introduced V(D)J recombination substrates by enforced RAG-1 and RAG-2 expression. We conclude that the Ku70 gene product is involved in DSBR and V(D)J recombination and confirm that the Ku70 gene can be classified as a member of the x-ray cross complementation group 6 (XRCC6). Potential differences between the Ku70(-/-) and Ku80(-/-) V(D)J recombination defects are discussed. PMID- 9223318 TI - Identification of immunodominant T cell epitopes of human glutamic acid decarboxylase 65 by using HLA-DR(alpha1*0101,beta1*0401) transgenic mice. AB - Glutamic acid decarboxylase isoform 2 (GAD65; EC 4.1.1.15) has been identified as a key target autoantigen of insulin-dependent diabetes mellitus (IDDM). IDDM is genetically associated with the major histocompatibility complex (MHC), and particular alleles from the HLA-DQ and HLA-DR loci contribute to disease. Among DR4 subtypes, HLA-DRB1*0401, HLA-DRB1*0402, and HLA-DRB1*0405 alleles lend susceptibility, while HLA-DRB1*0403 confers protection. We have utilized HLA DR(alpha1*0101,beta1*0401) (hereafter referred to as DR0401), human CD4, murine class II null triple transgenic mice and recombinant GAD65 to generate T cell hybridomas, and we have used overlapping sets of peptides to map the immunodominant epitopes of this autoantigen. We have identified 10 immunogenic regions for GAD65, of which 6 are recognized by multiple hybridomas. These epitopes are also generated by human antigen-presenting cells and their presentation is DR0401 restricted, as shown by the use of typed human lymphoblastoid cell lines and antibody blocking experiments. Immunodominant GAD65 epitopes defined in transgenic mice correspond to GAD65 regions previously shown to elicit T cell responses specifically in DR0401 IDDM patients, underscoring the validity of this approach. Interestingly, although the major epitopes contain DR0401 binding motifs, one of the epitopes contains a DR0405 motif. PMID- 9223319 TI - Natural killer cell acceptance of H-2 mismatch bone marrow grafts in transgenic mice expressing HLA-Cw3 specific killer cell inhibitory receptor. AB - Natural killer (NK) cells express killer cell inhibitory receptors (KIRs) for major histocompatibility complex class I molecules. Engagement of these surface receptors inhibits NK cell cytotoxic programs. KIR can also be expressed on T cell subsets, and their engagement similarly results in inhibition of effector functions initiated by the CD3/T cell receptor complex. KIR genes belong to two distinct families: the immunoglobulin superfamily (IgSF KIRs) and dimeric C2 lectins (lectin-like KIRs). Whereas both IgSF (p58: CD158, p70, and p140) and lectin-like KIRs (CD94/NKG2A heterodimers) have been found in human, only lectin like KIRs (all members of the Ly-49 family) have been described in the mouse. We have generated transgenic mice expressing an IgSF KIR, CD158b (p58.2), which recognizes HLA-Cw3. Our data show that CD158b is necessary and sufficient to confer specificity to NK cells, as well as to modulate T cell activation programs in vitro. In addition, we did not detect any adaptation of CD158b cell surface expression to that of HLA class I ligands in the CD158b x HLA-Cw3 double transgenic mice, in contrast to observations with Ly-49 in the mouse. Therefore, distinct strategies of selection/calibration appear to be used by IgSF and lectin like KIRs. Finally, the transgenic expression of CD158b KIR prevents the in vivo rejection of H-2 mismatch bone marrow grafts, which express the cognate major histocompatibility class I HLA-Cw3 allele, demonstrating for the first time the in vivo implication of human IgSF KIRs in the negative regulation of NK cell function. PMID- 9223320 TI - Characterization of tumor necrosis factor-deficient mice. AB - Although tumor necrosis factor (TNF) initially came to prominence because of its anti-tumor activity, most attention is now focused on its proinflammatory actions. TNF appears to play a critical role in both early and late events involved in inflammation, from localizing the noxious agent and amplifying the cellular and mediator responses at the local site and systemically, to editing (e.g., apoptosis) injured cells or effete immune cells and repairing inflammatory damage. We have generated mice deficient in TNF (TNF-/- mice) and have begun to examine the multiple functions attributed to TNF. TNF-/- mice develop normally and have no gross structural or morphological abnormalities. As predicted, they are highly susceptible to challenge with an infectious agent (Candida albicans), are resistant to the lethality of minute doses of lipopolysaccharide (LPS) following D-galactosamine treatment, have a deficiency in granuloma development, and do not form germinal centers after immunization. Phagocytic activity of macrophages appears relatively normal, as do T cell functions, as measured by proliferation, cytokine release, and cytotoxicity. B cell response to thymus independent antigens is normal, but the Ig response to thymus-dependent antigen is reduced. Surprisingly, cytokine production induced by LPS appears essentially intact, with the exception of reduced colony-stimulating factor activity. Other unexpected findings coming from our initial analysis are as follows. (i) TNF has low toxicity in TNF-/- mice. (ii) TNF-/- mice show an anomalous late response to heat-killed Corynebacterium parvum. In contrast to the prompt response (granuloma formation, hepatosplenomegaly) and subsequent resolution phase in C. parvum injected TNF+/+ mice, similarly treated TNF-/- mice show little or no initial response, but then develop a vigorous, disorganized inflammatory response leading to death. These results suggest that TNF has an essential homeostatic role in limiting the extent and duration of an inflammatory process-i.e., an anti inflammatory function. (iii) In contrast to the expectation that TNF+/+ mice and TNF+/- mice would have identical phenotypes, TNF+/- mice showed increased susceptibility to high-dose LPS lethality, increased susceptibility to Candida challenge, and delayed resolution of the C. parvum-induced inflammatory process, indicating a strong gene dose requirement for different actions of TNF. PMID- 9223321 TI - Manipulation of T cell costimulatory and inhibitory signals for immunotherapy of prostate cancer. AB - The identification of potentially useful immune-based treatments for prostate cancer has been severely constrained by the scarcity of relevant animal research models for this disease. Moreover, some of the most critical mechanisms involved in complete and proper antitumoral T cell activation have only recently been identified for experimental manipulation, namely, components involved in the costimulatory pathway for T cell activation. Thus, we have established a novel syngeneic murine prostate cancer model that permits us to examine two distinct manipulations intended to elicit an antiprostate cancer response through enhanced T cell costimulation: (i) provision of direct costimulation by prostate cancer cells transduced to express the B7.1 ligand and (ii) in vivo antibody-mediated blockade of the T cell CTLA-4, which prevents T cell down-regulation. In the present study we found that a tumorigenic prostate cancer cell line, TRAMPC1 (pTC1), derived from transgenic mice, is rejected by syngeneic C57BL/6 mice, but not athymic mice, after this cell line is transduced to express the costimulatory ligand B7.1. Also, we demonstrated that in vivo antibody-mediated blockade of CTLA-4 enhances antiprostate cancer immune responses. The response raised by anti CTLA-4 administration ranges from marked reductions in wild-type pTC1 growth to complete rejection of these cells. Collectively, these experiments suggest that appropriate manipulation of T cell costimulatory and inhibitory signals may provide a fundamental and highly adaptable basis for prostate cancer immunotherapy. Additionally, the syngeneic murine model that we introduce provides a comprehensive system for further testing of immune-based treatments for prostate cancer. PMID- 9223322 TI - Hypoxia-inducible factor-1 modulates gene expression in solid tumors and influences both angiogenesis and tumor growth. AB - Recent studies of tissue culture cells have defined a widespread system of oxygen regulated gene expression based on the activation of a heterodimeric transcription factor termed hypoxia-inducible factor-1 (HIF-1). To determine whether the HIF-1 transcriptional response is activated within solid tumors and to define the consequences, we have studied tumor xenografts of a set of hepatoma (Hepa-1) cells that are wild type (wt), deficient (c4), and revertant (Rc4) for an obligatory component of the HIF-1 heterodimer, HIF-1beta. Because HIF-1beta is also essential for the xenobiotic response (in which it is termed the aryl hydrocarbon receptor nuclear translocator), we also studied c31 cells, which have a different defect in the xenobiotic response and form the HIF-1 complex normally. Two genes that show different degrees of HIF-1-dependent hypoxia inducible expression in cell culture were selected for analysis-the glucose transporter, GLUT3, and vascular endothelial growth factor (VEGF). In situ hybridization showed intense focal induction of gene expression in tumors derived from wt, Rc4, and c31 cells, which was reduced (VEGF) or not seen (GLUT3) in those derived from c4 cells. In association with these changes, tumors of c4 cells had reduced vascularity and grew more slowly. These findings show that HIF 1 activation occurs in hypoxic regions of tumors and demonstrate a major influence on gene expression, tumor angiogenesis, and growth. PMID- 9223323 TI - Modified cytokeratins expressed on the surface of carcinoma cells undergo endocytosis upon binding of human monoclonal antibody and its recombinant Fab fragment. AB - Previously, we have reported on successful imaging of colon, rectal, and pancreatic carcinomas in patients by using a radiolabeled all-human monoclonal antibody, COU-1, directed against modified cytokeratin. To further develop this antibody for use as an immunoconjugate, COU-1 was cloned by phage display selection and the human Fab fragment was expressed in bacteria. Analysis by confocal laser scanning microscopy demonstrated that COU-1 bound in a uniform punctate pattern to the surface of viable carcinoma cells stained at 4 degrees C, and binding increased significantly when cells were cultured on fibronectin, laminin, or collagen IV. In the case of fibronectin, COU-1 staining was particularly enhanced at intercellular junctions. When carcinoma cells were cultured with COU-1 at 37 degrees C for 6 hr, the antibody was found in large perinuclear vesicles and the punctate surface staining was significantly reduced. Similar results were obtained using intact IgM COU-1 and the recombinant Fab fragment. Immunohistological studies indicated that COU-1, in contrast to murine monoclonal antibodies against normal cytokeratin 8 and 18, could differentiate between malignant and normal colon epithelia, and between colon cancer metastasis in the liver and surrounding normal hepatocytes. Within biopsies of malignant tissue, COU-1 exhibited membrane-associated staining of proliferating cells, while resting cells had a filamentous pattern. Thus, modified cytokeratin at the surface of carcinoma cells may represent a new target for immunoconjugates and may explain the promising results of the phase I/II clinical study. PMID- 9223324 TI - Tat protein induces self-perpetuating permissivity for productive HIV-1 infection. AB - We report that human immunodeficiency virus type 1 (HIV-1) has evolved a self perpetuating mechanism to actively generate cells permissive for productive and cytopathic infection. Only activated T cells can be productively infected, which leads to their rapid depletion (2 x 10(9)/day in an infected individual). Establishment of productive HIV-1 infection therefore requires continual activations from the large pool of quiescent T cells. Tat protein, which is secreted by infected cells, activated uninfected quiescent T cells in vitro and in vivo. These Tat-activated uninfected cells became highly permissive for productive HIV-1 infection. Activation of primary T cells by Tat protein involved integrin receptors and was associated with activation of mitogen-activated protein kinases, including ERK1 and JNK kinase. Accordingly, these primary T cells progressed from G0 to the late G1 phase of the cell cycle. PMID- 9223325 TI - Transgenic Galphaq overexpression induces cardiac contractile failure in mice. AB - The critical cell signals that trigger cardiac hypertrophy and regulate the transition to heart failure are not known. To determine the role of Galphaq mediated signaling pathways in these events, transgenic mice were constructed that overexpressed wild-type Galphaq in the heart using the alpha-myosin heavy chain promoter. Two-fold overexpression of Galphaq showed no detectable effects, whereas 4-fold overexpression resulted in increased heart weight and myocyte size along with marked increases in atrial naturietic factor ( approximately 55-fold), beta-myosin heavy chain ( approximately 8-fold), and alpha-skeletal actin ( approximately 8-fold) expression, and decreased ( approximately 3-fold) beta adrenergic receptor-stimulated adenylyl cyclase activity. All of these signals have been considered markers of hypertrophy or failure in other experimental systems or human heart failure. Echocardiography and in vivo cardiac hemodynamic studies indeed revealed impaired intrinsic contractility manifested as decreased fractional shortening (19 +/- 2% vs. 41 +/- 3%), dP/dt max, a negative force frequency response, an altered Starling relationship, and blunted contractile responses to the beta-adrenergic agonist dobutamine. At higher levels of Galphaq overexpression, frank cardiac decompensation occurred in 3 of 6 animals with development of biventricular failure, pulmonary congestion, and death. The element within the pathway that appeared to be critical for these events was activation of protein kinase Cepsilon. Interestingly, mitogen-activated protein kinase, which is postulated by some to be important in the hypertrophy program, was not activated. The Galphaq overexpressor exhibits a biochemical and physiologic phenotype resembling both the compensated and decompensated phases of human cardiac hypertrophy and suggests a common mechanism for their pathogenesis. PMID- 9223326 TI - Macrophages lacking scavenger receptor A show a decrease in binding and uptake of acetylated low-density lipoprotein and of apoptotic thymocytes, but not of oxidatively damaged red blood cells. AB - Macrophage binding of oxidatively damaged red blood cells (OxRBC) and apoptotic thymocytes correlates in many instances with a loss of phospholipid bilayer asymmetry, i.e., with an increase in expression of phosphatidylserine on the outer leaflet of the plasma membrane. Oxidatively modified LDL (OxLDL) can compete for the binding of these ligands to macrophages. However, the receptor(s) responsible remains to be identified. The present studies show that mouse peritoneal macrophages totally lacking scavenger receptor A (SRA) bound OxRBC just as effectively as wild-type macrophages, whereas their binding and uptake of acetyl LDL was reduced by more than 80%. Binding of apoptotic thymocytes and binding of OxLDL were also reduced, but only by 20-30%. We conclude that SRA is not involved in the recognition of phosphatidylserine-rich membranes but contributes to the binding of OxLDL and apoptotic thymocytes. The binding of OxRBC was almost totally calcium-dependent, whereas the binding of apoptotic thymocytes was not, suggesting that the mechanisms involved in their uptake by macrophages under these conditions were different. PMID- 9223327 TI - Altered expression of the WT1 wilms tumor suppressor gene in human breast cancer. AB - The product of the WT1 Wilms tumor suppressor gene controls the expression of genes encoding components of the insulin-like growth factor and transforming growth factor beta signaling systems. The role of these growth factors in breast tumor growth led us to investigate possible WT1 gene expression in normal and cancerous breast tissue. WT1 was detected by immunohistochemistry in the normal mammary duct and lobule, and the patterns of expression were consistent with developmental regulation. In a survey of 21 infiltrating tumors, 40% lacked immunodetectable WT1 altogether and an additional 28% were primarily WT1 negative. Cytoplasmic, but not nuclear, localization of WT1 was noted in some tumor cells and WT1 was detected, sometimes at high levels, in more-advanced estrogen-receptor-negative tumors. In this highly malignant subset, the tumor suppressor protein p53, which can physically interact with WT1, was also sometimes detected. WT1 mRNA was detected in normal and tumor tissue by reverse transcription-coupled PCR. Alternative splicing of the WT1 mRNA may regulate gene targeting of the WT1 protein through changes either in its regulatory or zinc finger domains. The relative proportions of WT1 mRNA splice variants were altered in a random sample of breast tumors, providing evidence that different tumors may share a common WT1-related defect resulting in altered regulation of target genes. PMID- 9223328 TI - Mouse model of GM2 activator deficiency manifests cerebellar pathology and motor impairment. AB - The GM2 activator deficiency (also known as the AB variant), Tay-Sachs disease, and Sandhoff disease are the major forms of the GM2 gangliosidoses, disorders caused by defective degradation of GM2 ganglioside. Tay-Sachs and Sandhoff diseases are caused by mutations in the genes (HEXA and HEXB) encoding the subunits of beta-hexosaminidase A. The GM2 activator deficiency is caused by mutations in the GM2A gene encoding the GM2 activator protein. For degradation of GM2 ganglioside by beta-hexosamindase A, the GM2 activator protein must participate by forming a soluble complex with the ganglioside. In each of the disorders, GM2 ganglioside and related lipids accumulate to pathologic levels in neuronal lysosomes, resulting in clinically similar disorders with an onset in the first year of life, progressive neurodegeneration, and death by early childhood. We previously have described mouse models of Tay-Sachs (Hexa -/-) and Sandhoff (Hexb -/-) diseases with vastly different clinical phenotypes. The Hexa /- mice were asymptomatic whereas the Hexb -/- mice were severely affected. Through gene disruption in embryonic stem cells we now have established a mouse model of the GM2 activator deficiency that manifests an intermediate phenotype. The Gm2a -/- mice demonstrated neuronal storage but only in restricted regions of the brain (piriform, entorhinal cortex, amygdala, and hypothalamic nuclei) reminiscent of the asymptomatic Tay-Sachs model mice. However, unlike the Tay Sachs mice, the Gm2a -/- mice displayed significant storage in the cerebellum and defects in balance and coordination. The abnormal ganglioside storage in the Gm2a -/- mice consisted of GM2 with a low amount of GA2. The results demonstrate that the activator protein is required for GM2 degradation and also may indicate a role for the GM2 activator in GA2 degradation. PMID- 9223329 TI - Thymineless death in colon carcinoma cells is mediated via fas signaling. AB - Fas is expressed constitutively in colonic epithelial cells and is also expressed in colon carcinomas and in cultured colon carcinoma cell lines. However, the potential role of Fas signaling in mediating apoptosis in cells of this type remains unknown. We have developed human colon carcinoma cell models deficient in thymidylate synthase that demonstrate acute (TS- cells) or delayed (Thy4 cells) apoptosis following DNA damage induced by thymineless stress. Complete protection of cells from acute apoptosis and prolongation of delayed apoptosis was obtained following exposure to the NOK-1 monoclonal antibody (inhibitory to Fas signaling) during the period of dThd deprivation. These results suggested that apoptosis induced by thymineless stress was regulated by autocrine signaling via Fas-FasL interactions. Fas expression was high in both TS- and Thy4 cells. However, FasL, undetectable in synchronous cultures, was up-regulated in TS- cells at 48 hr, when cells were undergoing acute apoptosis, and in Thy4 cells at 96 hr, correlating with the delayed onset of thymineless death. FasL expression also correlated with acute apoptosis induced in parental GC3/cl cells, commencing at 48 hr, following thymidylate synthase inhibition by 5-fluorouracil/leucovorin exposure. Fas-mediated apoptosis induced by the cytotoxic anti-Fas monoclonal antibody CH-11 was inhibited following adenoviral delivery of a Bcl-2 cDNA, and Bcl-2 also protected cells from acute apoptosis induced by dThd deprivation. Taken together, these data demonstrate a functional Fas system in these cultured colon carcinoma cell models, and they demonstrate that Fas-FasL interactions can link DNA damage induced by thymineless stress to the apoptotic machinery of colon carcinoma cells. PMID- 9223330 TI - The antisense bcl-2-IgH transcript is an optimal target for synthetic oligonucleotides. AB - In most human follicular B cell lymphomas the bcl-2 gene is up-regulated as a result of the t(14;18) chromosomal translocation generating a hybrid bcl-2-IgH mRNA. Recently, we have identified in t(14;18)-positive cells a bcl-2-IgH mRNA in the antisense orientation, putatively responsible for the overexpression of bcl 2. Herein we show that this chimeric antisense transcript is an optimal target for synthetic oligodeoxynucleotides (ODNs). A variety of sense-oriented oligonucleotides have been designed that target the antisense transcript in regions endowed with a sequence specificity presumably restricted to an individual cell line (the bcl-2-IgH fusion regions) or extended to all t(14;18) cells (the ectopic bcl-2 segment upstream from the major breakpoint region and the IgH segment). All sense-oriented ODNs complementary to the antisense transcript induced an early strong inhibition of cell growth and a late fulminant cell death. As expected, the activity of ODNs targeting the fusion region was restricted to each individual cell line, whereas the activity of all ODNs targeting the common bcl-2 and IgH segments was extended to all t(14;18) cell lines tested. These sense ODNs were not effective in untranslocated cell lines. Antisense-oriented ODNs, complementary to the bcl-2-IgH mRNA, and control ODNs (scrambled, inverted, or mismatched) were biologically ineffective. The selectivity and efficacy of all sense ODNs tested provide support for the development of therapeutic ODNs targeting the bcl-2-IgH antisense transcript expressed in human follicular lymphomas. PMID- 9223331 TI - Growth arrest of Epstein-Barr virus immortalized B lymphocytes by adenovirus delivered ribozymes. AB - Epstein-Barr virus (EBV) infection is associated with several human diseases that involve unrestricted proliferation of B lymphocytes. EBV nuclear antigen 1 (EBNA 1) is expressed in all EBV-infected cells and plays an essential role in persistence of the EBV genome. EBNA-1 has also been reported to have oncogenic potential. As an approach for treating EBV infections, we examined the capacity of EBNA-1 ribozymes delivered by recombinant adenoviruses to suppress EBNA-1 expression and to block virus-induced B cell proliferation. In contrast to primary B cells, EBV-transformed B lymphoblastoid cell lines expressed alphav integrins, the adenovirus internalization receptors, and were also susceptible to adenovirus-mediated gene delivery. Adenovirus delivery of a specific ribozyme (RZ1) to lymphoblastoid cell lines, suppressed EBNA-1 mRNA and protein expression, significantly reduced the number of EBV genomes, and nearly abolished cell proliferation in low serum. Adenovirus delivery of RZ1 also prevented EBV infection of an established EBV-negative B cell line. These studies demonstrate the potential use of adenovirus-encoded ribozymes to treat EBV-induced lymphoproliferative disorders. PMID- 9223333 TI - Gene expression from plasmids containing the araBAD promoter at subsaturating inducer concentrations represents mixed populations. AB - Gene expression from plasmids containing the araBAD promoter can be regulated by the concentration of arabinose in the growth medium. Guzman et al. [Guzman, L. M., Belin, D., Carson, M. J. & Beckwith, J. (1995) J. Bacteriol. 177, 4121-4130] showed that expression of a cloned gene could be modulated over several orders of magnitude in cultures grown in the presence of subsaturating concentrations of arabinose. We constructed plasmids expressing a fast-folding mutant Aequorea victoria green fluorescent protein from the araBAD promoter to examine the distribution of expressed gene products in individual cells at intermediate induction levels. Microscopic examination of cells grown at low arabinose concentrations shows mixtures of brightly fluorescent and dark cells, suggesting that intermediate expression levels in cultures reflect a population average of induced and uninduced cells. The kinetics of green fluorescent protein induction suggest that this reflects an "autocatalytic" induction mechanism due to accumulation of the inducer by active transport. This mechanism, which is analogous to the induction of the lac operon at subsaturating inducer concentrations in lacY+ cells, was described 40 years ago by Novick and Weiner [Novick, A. & Weiner, M. (1957) Proc. Natl. Acad. Sci. USA 43, 553-566]. PMID- 9223332 TI - The hepatitis B virus X gene induces p53-mediated programmed cell death. AB - The human hepatitis B virus (HBV) protein pX is a multifunctional regulatory protein that is known to affect both transcription and cell growth. Here we describe induction of apoptosis in NIH 3T3 polyclonal cell lines upon stimulation of pX expression from a dexamethasone inducible mouse mammary tumor virus (MMTV) X expression vector. The effect of long-term pX expression on the cell survival of mouse fibroblasts was confirmed in colony generation assays. This effect is not shared either by the other HBV products and it is c-myc mediated, as shown by the use of a dominant negative deletion mutant of c-myc. pX also sensitize cells to programmed cell death after exposure to DNA damaging agents. Taking advantage of stable transfectants carrying the p53val135 temperature-sensitive allele, we directly demonstrate that induction of apoptosis by pX requires p53. In p53 null mouse embryo fibroblasts pX activates transcription and confers an evident growth advantage without loss of cell viability. Although pX protein was not detectable in the experimental conditions we used, our results indicate that its expression affects both cell growth and cell death control. PMID- 9223334 TI - Acetylcholinesterase-transgenic mice display embryonic modulations in spinal cord choline acetyltransferase and neurexin Ibeta gene expression followed by late onset neuromotor deterioration. AB - To explore the possibility that overproduction of neuronal acetylcholinesterase (AChE) confers changes in both cholinergic and morphogenic intercellular interactions, we studied developmental responses to neuronal AChE overexpression in motoneurons and neuromuscular junctions of AChE-transgenic mice. Perikarya of spinal cord motoneurons were consistently enlarged from embryonic through adult stages in AChE-transgenic mice. Atypical motoneuron development was accompanied by premature enhancement in the embryonic spinal cord expression of choline acetyltransferase mRNA, encoding the acetylcholine-synthesizing enzyme choline acetyltransferase. In contrast, the mRNA encoding for neurexin-Ibeta, the heterophilic ligand of the AChE-homologous neuronal cell surface protein neuroligin, was drastically lower in embryonic transgenic spinal cord than in controls. Postnatal cessation of these dual transcriptional responses was followed by late-onset deterioration in neuromotor performance that was associated with gross aberrations in neuromuscular ultrastructure and with pronounced amyotrophy. These findings demonstrate embryonic feedback mechanisms to neuronal AChE overexpression that are attributable to both cholinergic and cell-cell interaction pathways, suggesting that embryonic neurexin Ibeta expression is concerted in vivo with AChE levels and indicating that postnatal changes in neuronal AChE-associated proteins may be involved in late-onset neuromotor pathologies. PMID- 9223335 TI - Intracisternal basic fibroblast growth factor enhances functional recovery and up regulates the expression of a molecular marker of neuronal sprouting following focal cerebral infarction. AB - Focal cerebral infarction (stroke) due to unilateral occlusion of the middle cerebral artery in mature rats produces deficits in sensorimotor function of the contralateral limbs that recover partially over time. We found that biweekly intracisternal injection of basic fibroblast growth factor (bFGF; 0.5 microg/injection), a potent neurotrophic polypeptide, markedly enhanced recovery of sensorimotor function of the contralateral limbs during the first month after stroke without apparent adverse side effects. Immunostaining for growth associated protein 43 (GAP-43), a molecular marker of axonal sprouting, showed a selective increase in GAP-43 immunoreactivity in the intact sensorimotor cortex contralateral to cerebral infarcts following bFGF treatment. These results show that bFGF treatment can enhance functional recovery after stroke, and that the mechanism may include stimulation of neuronal sprouting in the intact brain. PMID- 9223336 TI - Expression and activation of SH2/PTB-containing ShcA adaptor protein reflects the pattern of neurogenesis in the mammalian brain. AB - The adult mammalian brain comprises many functionally distinct neuronal types, which are generated during development as a result of a coordinated signaling cascade that drives neuroblasts from proliferation into differentiation. We investigated whether and how ShcA adaptor proteins, which are known to function as initiators of the Ras signaling cascade in various nonneuronal systems where they have been considered to be expressed ubiquitously, are involved in the proliferative and differentiative phases of the developing brain. We found that in the forebrain expression and activation of ShcA proteins were strictly regulated during embryonic development, both temporally and spatially. The mRNAs encoded by the ShcA gene were expressed exclusively within an area to which active proliferation of immature neuroblasts was confined, the ventricular zone. In postnatal and adult brain, ShcA mRNAs and proteins were present only faintly. In the adult olfactory epithelium, in which neuronal cell renewal occurs throughout life, ShcA remained strongly expressed. These phenomena were peculiar to ShcA, since Grb2 adaptor protein remained expressed at constant level throughout development. The embryonically expressed ShcA proteins were functionally active, since p52(ShcA) became phosphorylated on tyrosine and associated with Grb2 following intraventricular injection of epidermal growth factor in the embryonic brain. Our data indicate that, through an orderly pattern of expression, ShcA gene products may play a role in the control of the switch between proliferation and differentiation of brain neuroblasts. PMID- 9223337 TI - Brain-derived neurotrophic factor rapidly enhances phosphorylation of the postsynaptic N-methyl-D-aspartate receptor subunit 1. AB - Although neurotrophins have traditionally been regarded as neuronal survival factors, recent work has suggested a role for these factors in synaptic plasticity. In particular, brain-derived neurotrophic factor (BDNF) rapidly enhances synaptic transmission in hippocampal neurons through trkB receptor stimulation and postsynaptic phosphorylation mechanisms. Activation of trkB also modulates hippocampal long-term potentiation, in which postsynaptic N-methyl-D aspartate glutamate receptors play a key role. However, the final common pathway through which BDNF increases postsynaptic responsiveness is unknown. We now report that BDNF, within 5 min of exposure, elicits a dose-dependent increase in phosphorylation of the N-methyl-D-aspartate receptor subunit 1. This acute effect occurred in hippocampal synaptoneurosomes, which contain pre- and postsynaptic elements, and in isolated hippocampal postsynaptic densities. Nerve growth factor, in contrast, caused no enhancement of phosphorylation. These results suggest a potential mechanism for trophin-induced potentiation of synaptic transmission. PMID- 9223338 TI - Disruption of hippocampal development in vivo by CR-50 mAb against reelin. AB - We previously generated a monoclonal alloantibody, CR-50, by immunizing reeler mutant mice with homogenates of normal embryonic brains. This antibody recently was shown to recognize a Reelin protein, which is coded by the recently identified candidate gene for the reeler mutation. However, it is still unclear whether Reelin, especially the CR-50 epitope region, is indeed responsible for the reeler phenotype in vivo. Here we show that Reelin is localized on Cajal Retzius neurons in the hippocampus and that intraventricular injection of CR-50 at the embryonic stage disrupts the organized development of the hippocampus in vivo, converting it to a reeler pattern. Labeling experiments with 5 bromodeoxyuridine demonstrated that the labeled cells in the stratum pyramidale of the CR-50-treated mice were distributed in a pattern similar to that of reeler. Thus, Cajal-Retzius neurons play a crucial function in hippocampus development, and the CR-50 epitope on Reelin plays a central role in this function. PMID- 9223339 TI - Fixation of allosteric states of the nicotinic acetylcholine receptor by chemical cross-linking. AB - Receptor activity can be described in terms of ligand-induced transitions between functional states. The nicotinic acetylcholine receptor (nAChR), a prototypic ligand-gated ion channel, is an "unconventional allosteric protein" which exists in at least three interconvertible conformations, referred to as resting (low agonist affinity, closed channel), activated (open channel), and desensitized (high agonist affinity, closed channel). Here we show that 3,3'-dimethyl suberimidate (DMS) is an agonistic bifunctional cross-linking reagent, which irreversibly "freezes" the nAChR in a high agonist affinity/closed-channel state. The monofunctional homologue methyl acetoimidate, which is also a weak cholinergic agonist, has no such irreversible effect. Glutardialdehyde, a cross linker that is not a cholinergic effector, fixes the receptor in a low-affinity state in the absence of carbamoylcholine, but, like DMS, in a high-affinity state in its presence. Covalent cross-linking thus allows us to arrest the nAChR in defined conformational states. PMID- 9223340 TI - Interaction between amyloid precursor protein and presenilins in mammalian cells: implications for the pathogenesis of Alzheimer disease. AB - Mutations in the presenilin 1 (PS1) and presenilin 2 (PS2) genes increase the production of the highly amyloidogenic 42-residue form of amyloid beta-protein (Abeta42) in a variety of cell lines and transgenic mice. To elucidate the molecular mechanism of this effect, wild-type (wt) or mutant PS1 and PS2 genes were stably transfected into Chinese hamster ovary cells expressing endogenous or transfected beta-amyloid precursor protein (APP). By immunoprecipitation/Western blot analysis, APP was consistently found to coimmunoprecipitate with PS1 or PS2 proteins. Several distinct PS1, PS2, or APP antibodies precipitated PS-APP complexes that were detectable by blotting with either APP or PS antibodies. Importantly, complex formation could be detected at endogenous protein levels in nontransfected cells. In various Chinese hamster ovary cell lines, the amounts of APP coprecipitated by PS antibodies were proportional to the expression levels of both APP and PS. APP-PS complexes also were recovered from human 293 and HS683 cells. Full maturation of APP was not required for the interaction; most APP molecules complexed with PS were solely N-glycosylated. Treatment of cells with brefeldin A or incubation at 20 degrees C did not block complex formation, suggesting that the association between APP and PS occurs in part in the endoplasmic reticulum. Complex formation was detected for both wt and mutant PS and APP proteins. Deletion of the APP C-terminal domain did not abrogate complex formation, suggesting that the interaction does not occur in the cytoplasmic domains of the proteins. Our results demonstrate that wt and mutant PS1 and PS2 proteins form complexes with APP in living cells, strongly supporting the hypothesis that mutant PS interacts with APP in a way that enhances the intramembranous proteolysis of the latter by a gamma-secretase cleaving at Abeta42. PMID- 9223341 TI - Opioid receptors from a lower vertebrate (Catostomus commersoni): sequence, pharmacology, coupling to a G-protein-gated inward-rectifying potassium channel (GIRK1), and evolution. AB - The molecular evolution of the opioid receptor family has been studied by isolating cDNAs that encode six distinct opioid receptor-like proteins from a lower vertebrate, the teleost fish Catostomus commersoni. One of these, which has been obtained in full-length form, encodes a 383-amino acid protein that exhibits greatest sequence similarity to mammalian mu-opioid receptors; the corresponding gene is expressed predominantly in brain and pituitary. Transfection of the teleost cDNA into HEK 293 cells resulted in the appearance of a receptor having high affinity for the mu-selective agonist [D-Ala2, MePhe4-Gly-ol5]enkephalin (DAMGO) (Kd = 0.63 +/- 0.15 nM) and for the nonselective antagonist naloxone (Kd = 3.1 +/- 1.3 nM). The receptor had negligible affinity for U50488 and [D-Pen2, D Pen5]enkephalin (DPDPE), which are kappa- and delta-opioid receptor selective agonists, respectively. Stimulation of transfected cells with 1 microM DAMGO lowered forskolin-induced cAMP levels, an effect that could be reversed by naloxone. Experiments in Xenopus oocytes have demonstrated that the fish opioid receptor can, in an agonist-dependent fashion, activate a coexpressed mouse G protein-gated inward-rectifying potassium channel (GIRK1). The identification of six distinct fish opioid receptor-like proteins suggests that additional mammalian opioid receptors remain to be identified at the molecular level. Furthermore, our data indicate that the mu-opioid receptor arose very early in evolution, perhaps before the appearance of vertebrates, and that the pharmacological and functional properties of this receptor have been conserved over a period of approximately 400 million years implying that it fulfills an important physiological role. PMID- 9223342 TI - Functionally differentiating two neuronal nitric oxide synthase isoforms through antisense mapping: evidence for opposing NO actions on morphine analgesia and tolerance. AB - Several isoforms of neuronal nitric oxide synthase (nNOS) have been identified. Antisense approaches have been developed which can selectively down-regulate nNOS 1, which corresponds to the full-length nNOS originally cloned from the brain, and nNOS-2, a truncated form lacking two exons which is generated by alternative splicing, as demonstrated by decreases in mRNA levels. Antisense treatment also lowers nNOS enzymatic activity. Down-regulation of nNOS-1 prevents the development of morphine tolerance. Whereas morphine analgesia is lost in control and mismatch-treated mice given daily morphine injections for 5 days, mice treated with antisense probes targeting nNOS-1 show no decrease in their morphine sensitivity over the same time period. Conversely, an antisense probe selectively targeting nNOS-2 blocks morphine analgesia, shifting the morphine dose-response curve over 2-fold to the right. Both systems are active at the spinal and the supraspinal levels. An antisense targeting inducible NOS is inactive. Studies with NG-nitro-L-arginine, which does not distinguish among NOS isoforms, indicate that the facilitating nNOS-2 system predominates at the spinal level while the inhibitory nNOS-1 system is the major supraspinal nNOS system. Thus, antisense mapping distinguishes at the functional level two isoforms of nNOS with opposing actions on morphine actions. The ability to selectively down-regulate splice variants opens many areas in the study of nNOS and other proteins. PMID- 9223343 TI - Nitrergic control of peripheral sympathetic responses in the human corpus cavernosum: a comparison with other species. AB - Noradrenergic contractions induced by electrical field stimulation (EFS) of the rabbit anococcygeus muscle and the human and rabbit corpus cavernosum did not occur until termination of stimulation, even when EFS was applied for long periods (10 min). After treatment with a nitric oxide synthase inhibitor, a scavenger of NO, or a specific inhibitor of the soluble guanylate cyclase, EFS induced contraction began as soon as stimulation commenced and its magnitude and duration were increased. In the presence of a cGMP-phosphodiesterase inhibitor, the lag period between the end of EFS and the onset of contraction was longer, and the response was smaller. Even when the concentration of endogenous noradrenaline was increased with cocaine, the contraction still did not occur during EFS and the lag period was unchanged, although the response was enhanced. When tissue tone was elevated, relaxation occurred during EFS followed by a contraction. After blockade of neuronal noradrenaline release with guanethidine, contractions of the tissues to increasing concentrations of exogenous noradrenaline were significantly reduced by EFS, an effect that was reversible by inhibition of NO synthase. In contrast, in the rat and mouse anococcygeus muscles contraction began immediately with EFS, and nitrergic stimulation by EFS did not affect the responses elicited by high concentrations of exogenous noradrenaline. These results suggest that the human and rabbit genitourinary organs have a powerful nitrergic innervation that does not merely modulate, but actually controls, the sympathetic responses. Our observations may increase understanding of the balance between nitrergic and sympathetic systems in humans, disruption of which may contribute to certain pathological conditions. PMID- 9223344 TI - Quantitative trait loci controlling halothane sensitivity in Caenorhabditis elegans. AB - Genetic analysis is an essential tool for defining the molecular mechanisms whereby volatile anesthetics (VA) disrupt nervous system function. However, the degree of natural variation of the genetic determinants of VA sensitivity has not been determined nor have mutagenesis approaches been very successful at isolating significantly resistant mutant strains. Thus, a quantitative genetic approach was taken toward these goals. Recombinant-inbred strains derived from two evolutionarily distinct lineages of the nematode Caenorhabditis elegans were tested for sensitivity to clinically relevant concentrations (0.3-0.5 mM) of the VA halothane. The halothane sensitivities of coordinated movement and male mating behavior were highly variant among the recombinant-inbred strains with a range of EC50 values of 13- and 4-fold, respectively. Both traits were highly heritable (H2 = 0.82, 0.87, respectively). Several strains were found to be significantly resistant to halothane when compared with the wild-type strain N2. A major locus or loci mapping to the middle of chromosome V accounted for more than 40% of the phenotypic variance for both traits. Five weaker loci, four of which interact, explained most of the remaining variance. None of the halothane-sensitivity quantitative trait loci significantly affected behavior in the absence of halothane or halothane's potency for C. elegans immobilization, which requires 5 fold higher drug concentrations. Thus, the quantitative trait loci are unlikely to result from differences in halothane-independent (native) behavior or differences in halothane metabolism or permeability. Rather, these loci may code for targets and/or downstream effectors of halothane in the C. elegans nervous system or for modifiers of such gene products. PMID- 9223345 TI - Specific requirement of putrescine for the mitogenic action of juvenile hormone on adult insect neuroblasts. AB - Persistent neurogenesis in an adult insect brain was recently shown to be stimulated by juvenile hormone (JH). This morphogenetic hormone was also shown to act on polyamine biosynthesis. To analyze the possible involvement of polyamines in the neurogenic action of JH, two series of experiments were carried out with adult female crickets, Acheta domesticus: (i) inhibition of the first key enzyme in polyamine biosynthesis, ornithine decarboxylase, with alpha difluoromethylornithine (alpha-DFMO), and examination of the effects of this treatment on the neuroblast proliferation response to JH; and (ii) examination of the effects of putrescine supplementation on the mitotic index of JH-deprived and alpha-DFMO-treated females. In control females, alpha-DFMO treatment, as well as JH deprivation, greatly reduced neuroblast proliferation. Putrescine supplementation in alpha-DFMO-treated insects overcame the effects of alpha-DFMO, and allowed for detection of putrescine in the neural tissue and stimulation of brain neurogenesis. In JH-deprived females, alpha-DFMO treatment completely prevented the stimulatory action of JH on neuroblast proliferation and on brain putrescine levels. By contrast, putrescine feeding of JH-deprived animals was able to mimic the stimulatory effect of JH: brain putrescine levels increased and neuroblast proliferation was restored. To our knowledge, this report demonstrates for the first time that in vivo administration of putrescine can mimic the effects of a morphogenetic hormone on adult neuroblast proliferation, and shows the importance of polyamines, especially putrescine, in the transduction of JH message in neural tissue. PMID- 9223346 TI - AtMRP1 gene of Arabidopsis encodes a glutathione S-conjugate pump: isolation and functional definition of a plant ATP-binding cassette transporter gene. AB - Because plants produce cytotoxic compounds to which they, themselves, are susceptible and are exposed to exogenous toxins (microbial products, allelochemicals, and agrochemicals), cell survival is contingent on mechanisms for detoxifying these agents. One detoxification mechanism is the glutathione S transferase-catalyzed glutathionation of the toxin, or an activated derivative, and transport of the conjugate out of the cytosol. We show here that a transporter responsible for the removal of glutathione S-conjugates from the cytosol, a specific Mg2+-ATPase, is encoded by the AtMRP1 gene of Arabidopsis thaliana. The sequence of AtMRP1 and the transport capabilities of membranes prepared from yeast cells transformed with plasmid-borne AtMRP1 demonstrate that this gene encodes an ATP-binding cassette transporter competent in the transport of glutathione S-conjugates of xenobiotics and endogenous substances, including herbicides and anthocyanins. PMID- 9223347 TI - Cloning of a polycistronic cDNA from tomato encoding gamma-glutamyl kinase and gamma-glutamyl phosphate reductase. AB - We isolated from a tomato cDNA library the tomPRO1 locus, which encodes gamma glutamyl kinase (GK) and gamma-glutamyl phosphate reductase (GPR). This locus is unusual among eukaryotic genetic elements because it contains two open reading frames, and thus resembles prokaryotic polycistronic operons. The first open reading frame, specifying GK, is terminated by a TAA codon, which is followed by five nucleotides, an ATG translation initiation codon, and the second open reading frame, encoding GPR. DNA sequence analysis of fragments obtained by PCR amplification confirmed that the internal TAA and neighboring sequences are present in the endogenous tomPRO1 sequence in tomato. We demonstrated with RNase protection assays that the tomPRO1 locus is transcribed in tomato tissue culture cells, into a product that contains the internal stop codon. In Escherichia coli, tomPRO1 directed the synthesis of two proteins, a 33-kDa GK and a 44-kDa GPR. Antibodies against the 44-kDa GPR purified from E. coli recognized a 70-kDa product in tomato tissue culture cells and a 60-kDa product in leaves and roots. These results suggest that in tomato tissues, GPR is made as part of a longer polypeptide by some translational mechanism that enables bypass of the internal stop codon, such as frameshifting or ribosome hopping. The tomPRO1 locus may be the first example of a nuclear genetic element in plants that encodes two functional enzymes in two distinct open reading frames. PMID- 9223349 TI - Nontarget DNA sequences reduce the transgene length necessary for RNA-mediated tospovirus resistance in transgenic plants. AB - RNA-mediated virus resistance has recently been shown to be the result of post transcriptional transgene silencing in transgenic plants. This study was undertaken to characterize the effect of transgene length and nontarget DNA sequences on RNA-mediated tospovirus resistance in transgenic plants. Transgenic Nicotiana benthamiana plants were generated to express different regions of the nucleocapsid (N) protein of tomato spotted wilt (TSWV) tospovirus. Transgenic plants expressing half-gene segments (387-453 bp) of the N gene displayed resistance through post-transcriptional gene silencing. Although smaller N gene segments (92-235 bp) were ineffective in conferring resistance when expressed alone in transgenic plants, these segments conferred resistance when fused to the nontarget green fluorescent protein gene DNA. These results demonstrate that (i) a critical length of N transgene (236-387 bp) is required for a high level of transgene expression and consequent gene silencing, and (ii) the post transcriptional gene silencing mechanism can trans-inactivate the incoming tospovirus genome with homologous transgene segments that are as short as 110 bp. Therefore, the activation of post-transcriptional transgene silencing requires a significantly larger transgene than is required for the trans-inactivation of the incoming viral genome. These results raise the possibility of developing a simple new strategy for engineering multiple virus resistance in transgenic plants. PMID- 9223348 TI - Inheritance, gene expression, and lignin characterization in a mutant pine deficient in cinnamyl alcohol dehydrogenase. AB - We have discovered a mutant loblolly pine (Pinus taeda L.) in which expression of the gene encoding cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) is severely reduced. The products of CAD, cinnamyl alcohols, are the precursors of lignin, a major cell wall polymer of plant vascular tissues. Lignin composition in this mutant shows dramatic modifications, including increased incorporation of the substrate of CAD (coniferaldehyde), indicating that CAD may modulate lignin composition in pine. The recessive cad-n1 allele, which causes this phenotype, was discovered in a tree heterozygous for this mutant allele. It is inherited as a simple Mendelian locus that maps to the same genomic region as the cad locus. In mutant plants, CAD activity and abundance of cad RNA transcript are low, and free CAD substrate accumulates to a high level. The wood of the mutant is brown, whereas the wood in wild types is nearly white. The wood phenotype resembles that of brown midrib (bm) mutants and some transgenic plants in which xylem is red brown due to a reduction in CAD activity. However, unlike transgenics with reduced CAD, the pine mutant has decreased lignin content. Wood in which the composition of lignin varies beyond previous expectations still provides vascular function and mechanical support. PMID- 9223351 TI - Role of endothelial dysfunction in coronary artery disease. AB - The vascular endothelium plays an essential role in regulating blood flow and other functions of the coronary arteries. Under normal conditions, the endothelium releases a number of factors that regulate arterial vasomotion. One of these factors, endothelial-derived relaxing factor (EDRF), is a vasorelaxant that is identical to, or closely related to, nitric oxide (NO). Endothelial function may be compromised in coronary artery disease (CAD) or in the presence of risk factors for CAD. In this setting, EDRF-NO activity is inhibited, which may lead to vasoconstriction, platelet aggregation, vasospasm, and thrombosis. The relation between endothelial function, arterial vasomotion, and myocardial ischemia is discussed, with particular emphasis on the role of EDRF-NO, and the therapeutic interventions that may be useful in treating the clinical consequences of endothelial dysfunction. PMID- 9223350 TI - Integration of local pattern elements into a global shape in human vision. AB - The spatial extent of the cortical filters selective for different spatial frequencies and orientations is limited. We studied psychophysically how information from the local filters is integrated into global pattern shapes, i.e., whether performance in the identification of a global pattern consisting of small, locally oriented Gabor elements depends on the orientations of those elements. The observer was presented with an E-like stimulus pattern shape comprised of oriented Gabor patches on a blank background, and the performance measure was the threshold contrast for identifying the orientation of the E pattern (four possible rotated orientations). The results showed that contrast thresholds were significantly lower when the local elements all shared the same orientation (e.g., all horizontal) compared with the condition in which the elements had mixed orientations (both horizontal and vertical). The enhancement effect due to uniform local orientations can be explained by two factors: One is local facilitatory interactions between the orientation selective filters, and the other is second-order information integration across the filters. PMID- 9223352 TI - Risk stratification in coronary artery disease: implications for stabilization and prevention. AB - Noninvasive nuclear imaging techniques, including dual-isotope myocardial perfusion single-photon emission computed tomography (SPECT), have been employed in the development of strategies for diagnosis and risk stratification of patients with suspected or known coronary artery disease. These risk stratification strategies are based on studies in which known outcome has been linked to diagnostic and prognostic information provided by myocardial perfusion SPECT. This article describes a validated dual-isotope exercise protocol for assessment of perfusion and function and reviews the evidence on which a cost effective risk management strategy is based. PMID- 9223353 TI - New insights into the pathogenesis and prevention of acute coronary syndromes. AB - Several recent studies have shown that 60-70% of coronary occlusions that cause acute coronary syndromes (such as unstable angina, myocardial infarction, or sudden ischemic death) evolve from atherosclerotic plaques that are only mildly to moderately obstructive. Numerous studies have demonstrated that coronary thrombosis, the immediate cause of acute coronary syndromes, is a consequence of plaque disruption. Most thrombotic events are related to deep plaque fissure, while superficial plaque erosion is the cause in a significant minority of cases. Thus, the mechanisms by which stable coronary artery disease (CAD) evolves into an unstable and potentially lethal acute coronary syndrome are related to plaque disruption and thrombosis. The vulnerability of a plaque to disruption appears to be determined by the presence of a large lipid-rich core, a thin fibrous cap, and an inflammatory cellular infiltrate, rather than by the size of the plaque or the severity of stenosis caused by a plaque before disruption. In addition to plaque disruption and thrombosis, enhanced vasoconstriction--a characteristic feature of CAD and dyslipidemia-may contribute to the clinical manifestations of CAD. Angiographic studies have demonstrated that risk factor modification produces a disproportionately greater reduction in ischemic clinical events than in anatomic regression of plaque, suggesting "plaque stabilization" may be the major mechanism of such clinical benefit. The relatively rapid attenuation of endothelial-mediated vasomotor dysfunction with the treatment of dyslipidemia lends credence to this concept. PMID- 9223354 TI - Management of patients with chronic stable angina at low risk for serious cardiac events. AB - Successful management of the patient with chronic stable angina requires correct stratification by assessing the risk of future coronary events. Patients at low risk for such events have a relatively good prognosis; revascularization procedures (balloon angioplasty or surgery) offer no benefit over medical management. Such patients should be offered medical therapy as their first option. The goals in management of chronic stable angina are (1) treatment of other conditions that may worsen angina; (2) treatment with aspirin and modification of risk factors for coronary artery disease (CAD) to improve outcome; and (3) effective relief of anginal symptoms. Most patients with stable angina will have CAD. It is well established that treatment with aspirin and modification of risk factors for CAD are beneficial in reducing cardiovascular mortality and morbidity. Blood pressure reduction, lowering of blood cholesterol level, and smoking cessation are interventions of proven value and appear to correct defects (at least partially) in the endothelial function of the coronary blood vessels. Other interventions that are helpful are estrogen replacement treatment in postmenopausal women, and low-dose aspirin therapy-which is recommended for all patients who can tolerate it. For controlling symptoms and improving angina-free walking time, nitrates, beta blockers, and calcium channel antagonists are efficacious as first-line monotherapy for chronic stable angina in this group of patients. Nitrates may be of special use in patients with impaired left ventricular function, overt congestive heart failure, intermittent coronary vasoconstriction, or coronary artery spasm. In patients with concomitant hypertension or supraventricular tachycardia, beta blockers are helpful. Calcium channel antagonists may be useful in patients with chronic obstructive pulmonary disease, peripheral vascular disease, or hypertension. When optimal monotherapy with a given class of drug fails to control symptoms, alternative monotherapy with a different class of agent should be tried before combination therapy. Combination therapy with 2 or 3 agents is not always superior to optimal monotherapy. Patients who fail to respond to adequate medical therapy should be considered for a revascularization procedure. PMID- 9223355 TI - Achieving sustained improvement in myocardial perfusion: role of isosorbide mononitrate. AB - The efficacy of antianginal agents in the treatment of patients with chronic stable angina has traditionally been evaluated by performance measures, such as the exercise treadmill test (ETT). Although reliable and reproducible, ETT is not a sensitive measure of changes in myocardial ischemia. The effects of antianginal agents on coronary blood flow and myocardial perfusion have been less frequently studied. Angiographic studies have demonstrated that nitrates may operate by preferentially directing blood flow to ischemic regions of the myocardium. These investigations have been limited, however, by the invasive nature of the evaluation. Measurements of regional myocardial perfusion may also be made with noninvasive tests. Both quantitative single-photon emission computed tomography (SPECT) and positron emission tomography (PET) have been employed, but few studies have used these techniques to assess the effects of antianginal drugs (in general) and nitrates (in particular) on changes in reversible myocardial perfusion defects. Studies that have evaluated the direct effects of nitrate treatment on coronary blood flow and myocardial perfusion defects in patients with chronic stable angina are reviewed, and preliminary data from a study of the effects of long-term nitrate treatment on myocardial perfusion are discussed. PMID- 9223356 TI - Adenosine and adenosine receptors in the cardiovascular system: biochemistry, physiology, and pharmacology. AB - Cardiomyocytes and vascular cells readily form, transport, and metabolize the endogenous adenine nucleoside adenosine and act to regulate both interstitial and plasma adenosine concentrations. Cardiovascular cells also have membrane adenosine receptors. Cell and tissue distributions, signal transduction pathways, and pharmacology of each of the four subtypes of adenosine receptors are subjects of intense investigation. The A1-adenosine receptors mediate the negative dromotropic, chronotropic, inotropic, and the anti-beta-adrenergic actions of adenosine. Activation of A(2A)- and perhaps A(2B)-adenosine receptors causes vasodilation. Evidence of novel actions mediated by A(2B)- and A3-adenosine receptors is accumulating. Adenosine is cardioprotective during episodes of cardiac hypoxia/ischemia; several potential mechanisms may be involved. Pharmacologic tools are currently available for laboratory investigation of the actions of adenosine, and the development of adenosine receptor subtype-selective agonists and antagonists for therapeutic purposes is beginning. PMID- 9223357 TI - Autonomic neural control of cardiac function: modulation by adenosine and adenosine 5'-triphosphate. AB - Adenosine and adenosine 5'-triphosphate (ATP) are found in every cell of the human body. These molecules are released from cells into the extracellular fluid under physiologic and pathophysiologic conditions. Outside of cells, adenosine and ATP act as physiologic regulators of cells, tissues, and organs. In the heart, extracellular adenosine and ATP exert pronounced inotropic, lusitropic, electrophysiologic, and metabolic effects, which are mediated by specific cell surface receptors. In addition, both compounds can modulate sympathetic and parasympathetic input to the heart by interacting with neural elements within and without the heart, thereby modulating autonomic neural control of cardiac functions. This article briefly reviews these indirect, neurally-mediated actions of adenosine and ATP. PMID- 9223358 TI - Role of adenosine in the cardiac catheterization laboratory. PMID- 9223359 TI - Myocardial perfusion imaging during adenosine-induced coronary hyperemia. PMID- 9223360 TI - Adenosine stress echocardiography. PMID- 9223361 TI - Adenosine as an antiarrhythmic agent. AB - Adenosine produces acute inhibition of sinus node and atrioventricular (AV) nodal function. This profound but short lived electrophysiologic effect makes adenosine a suitable agent for treating supraventricular tachycardias (SVT) that incorporate the sinus node or AV node as part of the arrhythmia circuit, or for unmasking atrial tachyarrhythmias or ventricular pre-excitation. Its antiadrenergic properties also make it an effective agent for use with some unique atrial and ventricular tachycardias. Appropriate dosing and rapid bolusing with intravenous administration is required. Recognition of infrequent proarrhythmic risks and potential drug interactions with xanthine derivatives and dipyridamole should maximize its safe and effective use. This review will highlight adenosine's mechanism of action, administration, clinical indications, efficacy, and risks when used in tachyarrhythmic management. PMID- 9223362 TI - Safety, tolerance, and efficacy of adenosine as an additive to blood cardioplegia in humans during coronary artery bypass surgery. AB - Myocardial stunning after heart surgery is associated with increased morbidity and mortality in patients with severe multivessel disease and reduced myocardial function. The purpose of this study was to evaluate the safety, tolerance, and efficacy of adenosine as a cardioprotective agent when added to blood cardioplegia in patients undergoing coronary artery bypass surgery. Sixty-one patients were randomized to standard cold-blood cardioplegia, or cold-blood cardioplegia containing 1 of 5 adenosine doses (100 microM, 500 microM, 1 mM, 2 mM, and 2 mM with a preischemic infusion of 140 microg/kg/min of adenosine). Invasive and noninvasive measurements of ventricular performance and rhythm were obtained preoperatively, prebypass, and then at 1, 2, 4, 8, 16, and 24 hours postbypass. Use of inotropic agents and vasoactive drugs pastoperatively was recorded; blood samples were collected for measurement of nucleoside levels. High dose adenosine treatment was associated with a 249-fold increase in the plasma adenosine concentration and a 69-fold increase in the combined levels of adenosine, inosine, and hypoxanthine (p <0.05). Increasing doses of the adenosine additive were also associated with lower requirements of dopamine (p = 0.003) and nitroglycerine (p = 0.001). The 24-hour average doses for dopamine and nitroglycerine in the placebo group were 28-fold and 2.6-fold greater than their respective high-dose adenosine treatment cohorts. Finally, the placebo- and 100 microM-adenosine group was associated with a lower ejection fraction when compared to patients receiving the intermediate dose or high-dose treatment. These findings are consistent with the hypothesis that adenosine is effective in attenuating myocardial stunning in humans. PMID- 9223363 TI - Adenosine for myocardial protection in acute myocardial infarction. AB - Although reperfusion therapy for acute myocardial infarction is known to reduce infarct size, improve left ventricular function, and reduce mortality, the full potential benefit may be limited by acceleration of damage resulting from reperfusion, or "reperfusion injury." Evidence of a variety of mechanisms of reperfusion injury has led to a wide range of proposed therapeutic interventions, including agents to prevent oxygen free radical damage, inhibit white blood cell function, reduce calcium influx, improve microvascular blood flow, inhibit sympathetic stimulation, and improve energy stores. A multitude of agents have been shown to limit infarct size in animals when administered before or during reperfusion. Unfortunately, most have been disappointing when tested clinically. Adenosine, a theoretically attractive agent for preventing reperfusion injury, has shown promise in small, clinical studies, and appears to be an endogenous substance involved in the protective effect of ischemic preconditioning. When studied in the setting of angioplasty for acute myocardial infarction, adenosine was associated with small infarct size and improved coronary flow. As myocardial preservation with reperfusion during bypass surgery shares pathophysiologic characteristics with the reperfused myocardium in acute infarction, early results of adenosine during bypass surgery presented at this symposium support the concept that adenosine may be beneficial. Two ongoing Phase II trials of adenosine in acute myocardial infarction-one with thrombolysis and one with direct angioplasty-will provide important information about the potential benefits of adenosine in the context of reperfusion. PMID- 9223364 TI - Interaction of epidermal growth factor and insulin-like growth factor-I in the regulation of growth plate chondrocytes. AB - The action of growth factors on the cells of the epiphyseal growth plate is an important mechanism in the regulation of skeletal growth. Insulin-like growth factor-I (IGF-I) is known to play a central role in the regulation of bone growth. In contrast, the role, if any, of epidermal growth factor (EGF) is not yet clear. In these studies, we tested the hypothesis that EGF interacts with IGF I in the regulation of growth plate chondrocyte mitotic and metabolic activities. Chondrocytes isolated from bovine radioulnar growth plates and incubated in suspension culture were analyzed for their responsiveness to EGF with respect to synthesis of DNA, proteins, and proteoglycans, responsiveness to IGF-I, and ability to specifically bind [125I]IGF-I. Treatment of growth plate chondrocytes with maximally effective concentrations (10-100 ng/ml) of EGF produced a 16-27% increase in specific binding of [125I]IGF-I. Scatchard analysis indicated that this increase in specific binding was due to an increase in the number of receptors/cell with no change in receptor affinity. EGF stimulated protein synthesis by 30-35%. Pretreatment with EGF increased the responsiveness of chondrocytes to IGF-I, resulting in 90 and 60% augmentation of IGF-I-stimulated mitotic activity and proteoglycan synthesis, respectively. Given the prominent role of IGF-I in skeletal development and the presence of EGF in the growth plate, this study suggests an important role for interactions between these growth factors in the regulation of skeletal growth. PMID- 9223365 TI - Injury stimulates outgrowth and motility of oligodendrocytes grown in vitro. AB - Oligodendrocytes which form myelin within the CNS develop from small, highly motile cells that are largely bipolar into mature cells which extend many processes and which produce myelin membranes around multiple axons. The production of myelin sheaths is thought to anchor mature oligodendrocytes (OLs), limiting their motility. When the brain sustains an injury, OLs do not make a significant effort to remyelinate, a fact attributed to both their lack of proliferation and their inability to migrate or extend processes into areas of injury. To test the motility and growth potential of mature OLs, we have designed an in vitro system in which individual cells can undergo long-term observation. Additionally, cells can be mechanically injured by transection of processes using a low-power laser beam. Both control and injured OLs undergo several types of structural change, including extension and retraction of processes and membranes, as well as changes in process caliber. Some OLs exhibit a high degree of motility, moving several hundred micrometers within days. Rather than interfering with the cells' ability to undergo structural change, injury actually stimulated outgrowth of new processes and motility. Neither injury nor addition of basic fibroblast growth factor (bFGF) increased the rate of OL division. However, bFGF paradoxically caused an increase in uptake of the DNA synthesis marker bromodeoxyuridine and had negative effects on OL survival. The unexpected findings that OLs with a mature phenotype are motile and undergo constant structural modification in vitro and that injury induces certain behaviors suggest that myelin-forming OLs in the brain may be capable of a high degree of plasticity under certain conditions. PMID- 9223366 TI - Regulation of cellular tyrosine phosphorylation by stimulatory and inhibitory muscarinic acetylcholine receptors. AB - Tyrosine phosphorylation is a key signaling event in transmembrane and cytoplasmic signal transduction. The m5 muscarinic receptor (m5AChR) responds to ligand stimulation with calcium influx and protein phosphorylation. In contrast, neither of these responses has been associated with m4AChR signaling. We hypothesized that activation of the m5AChR would alter tyrosine phosphorylation patterns spatially within the cell and in a calcium influx-sensitive manner. CHO cells stably transfected with m4- or m5AChRs were assessed for spatial localization and quantity of phosphotyrosylated proteins in response to receptor activation. Results were confirmed by immunoblot of whole cell lysates and cytosol and membrane fractions. m5AChR activation increased tyrosine phosphorylation in all subcellular compartments; coincubation with CAI, a calcium influx inhibitor, reduced phosphorylation below basal levels. Western blot confirmed the change of phosphotyrosylated proteins of M(r) 70, 85, 120, and 180 kDa in whole and fractionated cells. PLC-gamma, used as a marker of m5AChR activity, was increased in quantity and degree of phosphorylation in CHOm5 cell membranes and microvilli in response to receptor activation. Both the quantitative increase and tyrosine phosphorylation of PLC-gamma in membrane fractions was inhibited by CAI. In contrast, CC treatment of CHOm4 cells reduced tyrosine phosphorylation throughout the cell. CC-stimulation of m5AChR cells caused a calcium influx-sensitive increase in phosphotyrosylated proteins throughout the cell, though predominantly in the membrane and microvilli. Activation of the m5AChR induces tyrosine phosphorylation, whereas activation of the m4AChR inhibited tyrosine phosphorylation below baseline, further demonstrating the dichotomy between signaling of these two AChRs. PMID- 9223367 TI - Cytoplasmic localization of cyclin D3 in seminiferous tubules during testicular development. AB - Using a newly developed polyclonal antibody against murine cyclin D3, we have found that protein levels of cyclin D3 were highly detectable only in thymus and testis in rats. Since testis offer unique opportunities to examine the cell cycle in vivo, we examined the temporal and spatial expression of cyclin D3 and the DNA synthesis indicator, proliferating cell nuclear antigen (PCNA), in the rat testis during development. The protein levels of cyclin D3 protein in testis from 7 days to 3 months old were almost constant and then decreased gradually thereafter. The protein levels of cyclin D1 and PCNA were high in the testis of 7- and 14-day-old rats and decreased during testicular development. In the seminiferous tubules of 7-day-old newborns, cyclin D3 was surprisingly located in cytoplasm of stem cells that had bigger nuclei than the nuclei of surrounding cells. Interestingly, cyclin D3 immunopositive cells did not immunostain with PCNA in nuclei. In the adult testis, anti-cyclin D3 antibody strongly stained the cytoplasm of early stage primary spermatocytes, lightly stained pachytene spermatocytes, but did not stain elongated spermatids. There was no detectable cyclin D3 in Sertoli cells, interstitial cells, or fibroblasts within seminiferous tubules, or in blood vessels within the interstitial matrix. The known cyclin D3 partner, cyclin dependent kinase 4, was located mainly in nuclei of spermatogonia and in early stage primary spermatocytes. Strong PCNA immunopositive staining was located in the nuclei of spermatogonia in adult testis. These results indicate that cyclin D3 is detectable in meiotically active male germ cells (PCNA-negative cells), but is conspicuously absent from mitotically active spermatogonia (PCNA-positive cells). Moreover, in contrast to in vitro reports, cyclin D3 is not located in the nucleus, but rather in the cytoplasm of male germ cells in vivo. Taken together, the presence of cyclin D3 in spermatocytes and its location in the cytoplasm lead us to speculate that cyclin D3 may have functions in male germ cells other than mitosis. PMID- 9223368 TI - Induction of apoptosis through the PKC pathway in cultured dermal papilla fibroblasts. AB - The dermal papilla (DP) consists of a discrete population of specialized fibroblasts that are important in the morphogenesis of the hair follicle in the embryo and in the control of the hair growth cycle in the adult. This mitotically quiescent and long-lived cell population expresses gene products that promote cell survival such as Bcl-2, and thus normally might be protected from apoptosis. We investigated whether cultured DP fibroblasts are able to undergo apoptosis by treatment with the protein kinase inhibitor staurosporine. Involvement of the PKC signaling pathway in DP fibroblast survival/death was investigated by inhibition (staurosporine and Bisindolylmaleimide (Bis) treatment) or activation (TPA; 12-O tetradecanoylphorbol-13-acetate treatment) of PKC and characterization of DP expressed PKC isoforms by RT-PCR. We determined that cultured DP fibroblasts undergo apoptosis, in a dose-related manner, when treated with staurosporine but not when treated with Bis, an inhibitor with narrow PKC isoform specificity. TPA confers partial and transient resistance to staurosporine-induced DP apoptosis. Staurosporine and Bis each induced G1 arrest, whereas TPA treatment of cultured DP resulted in increased entry into S-phase. The differential responses to individual inhibitors and activators of PKC may be related to the multiple PKC isoforms that DP fibroblasts express. Flow cytometric analysis indicates that the mechanism of staurosporine-induced apoptosis may be through decrease of Bcl-2 in treated DP cells or through modulation of cell cycle regulators. Correlation between sensitivity to induction of apoptosis and proliferation suggests that dermal papilla cells may normally be protected from apoptosis in vivo by their mitotically quiescent state. PMID- 9223369 TI - Sensitivity of transformed fibroblasts for intercellular induction of apoptosis is determined by their transformed phenotype. AB - Intercellular induction of apoptosis defines a potential control mechanism of oncogenesis. It is based on induction of apoptosis in transformed fibroblasts by neighboring nontransformed fibroblasts. Transforming growth factor type beta (TGF beta) represents the initial triggering molecule to induce nontransformed cells to release apoptosis-inducing factors. To test whether sensitivity for intercellular induction of apoptosis is directly dependent on the transformed phenotype, v-src-transformed rat fibroblasts and emerging revertants were tested for their sensitivity. All transformed cell clones were sensitive, whereas all revertant clones had lost their sensitivity in parallel with the loss of the transformed phenotype. In addition, revertants had regained the potential to induce apoptosis in transformed cells. Sensitivity to intercellular induction of apoptosis is therefore directly dependent on the transformed phenotype, whereas the ability to induce apoptosis is a specific feature of nontransformed fibroblasts. PMID- 9223370 TI - p53 Induces myocyte apoptosis via the activation of the renin-angiotensin system. AB - The mechanism by which p53 activates apoptosis in various cell systems is unknown. In the absence of an external death stimulus, p53 and p53-dependent genes, bcl-2 and bax, cannot trigger apoptosis. However, p53 may enhance not only transcription of bax and repress bcl-2, but also may upregulate the local renin angiotensin system, inducing the formation and secretion of angiotensin II from the cells. To test this hypothesis, adult rat ventricular myocytes were infected with AdCMV.p53, which resulted in downregulation of Bcl-2, upregulation of Bax, and death of 34% of the cells. Gel retardation assays demonstrated p53 binding in the promoters of angiotensinogen and angiotensin II AT1 receptor subtype. Angiotensinogen and AT1 mRNAs increased in AdCMV.p53 cells and this phenomenon was associated with a 14-fold increase in the secretion of angiotensin II. The AT1 receptor blocker losartan and angiotensin II antibody prevented p53-induced apoptosis. Thus, p53 enhances the myocyte renin-angiotensin-system and decreases the Bcl-2/Bax ratio in the cells, triggering apoptosis. The identification of this new pathway in p53-mediated apoptosis may be critical in the alterations of myocardial function in the pathologic heart. PMID- 9223371 TI - Characterization of a mutant profilin with reduced actin-binding capacity: effects in vitro and in vivo. AB - We are investigating structure-function relationships in profilin and actin by site-specific mutagenesis using a yeast, Saccharomyces cerevisiae, expression system to produce wild-type and mutant proteins. This paper shows that deleting proline 96 and threonine 97, which are located close to the major actin binding site on profilin, did not significantly alter the interaction between profilin and phosphatidylinositol 4,5-bisphosphate, nor did it affect the profilin:poly(L proline) interaction. The mutant protein, however, had a lower capacity to bind to actin in vitro than wild-type profilin, though it showed a slightly increased profilin-enhanced nucleotide exchange on the actin. When microinjected into Swiss 3T3 mouse fibroblasts or porcine aortic endothelial cells, the mutant profilin did not change the organization of the microfilament system like the wild-type profilin did. This provides further evidence that profilin controls microfilament organization in the cell by interacting directly with actin. PMID- 9223372 TI - Paraquat inhibits the processing of human manganese-dependent superoxide dismutase by SF-9 insect cell mitochondria. AB - Precursor human manganese-dependent superoxide dismutase (hMn-SOD) was expressed using the baculovirus system in Spodoptera fungiperda (Sf-9) insect cells. Following infection of Sf-9 cells with hMn-SOD-expressing baculovirus, mature hMn SOD was expressed at 15-25% of total cellular protein, with the recombinant protein localized in the mitochondrial matrix. Partial amino acid sequencing and SOD activity assays indicated that the hMn-SOD was correctly processed and assembled by insect mitochondria. This expression system was used to study the effects of paraquat and menadione, two intracellular superoxide generators, on processing of precursor hMn-SOD by insect mitochondria. Paraquat was found to potently inhibit mitochondrial processing of hMn-SOD, leading to the accumulation of precursor hMn-SOD and a decrease in measurable Mn-SOD activity. In contrast, menadione treatment was not found to affect the ratio of precursor to mature Mn SOD. Paraquat did not lead to lower total production of hMn-SOD or cellular toxicity at the concentrations which were found to block processing of precursor hMn-SOD. These results indicate that mitochondrial processing and import of the precursor protein hMn-SOD are early events susceptible to dysfunction induced by the redox-cycling agent paraquat. These results also emphasize that mitochondrial processing of precursor proteins represents a parameter of cellular function which may be compromised, preceding the appearance of more generalized deterioration of cellular function, under certain toxic or pathological conditions. PMID- 9223373 TI - Lysosomal biogenesis in lysosomal storage disorders. AB - Lysosomal biogenesis is an orchestration of the structural and functional elements of the lysosome to form an integrated organelle and involves the synthesis, targeting, functional residence, and turnover of the proteins that comprise the lysosome. We have investigated lysosomal biogenesis during the formation and dissipation of storage vacuoles in two model systems. One involves the formation of sucrosomes in normal skin fibroblasts and the other utilizes storage disorder-affected skin fibroblasts; both of these systems result in an increase in the size and the number of lysosomal vacuoles. Lysosomal proteins, beta-hexosaminidase, alpha-mannosidase, N-acetylgalactosamine-4-sulfatase, acid phosphatase, and the lysosome-associated membrane protein, LAMP-1, were shown to be elevated between 2- and 28-fold above normal during lysosomal storage. Levels of mRNA for the lysosome-associated membrane proteins LAMP-1 and LAMP-2, N acetylgalactosamine-4-sulfatase, and the 46- and 300-kDa mannose-6-phosphate receptors were also elevated 2- to 8-fold. The up-regulation of protein and mRNA lagged 2-4 days behind the formation of lysosomal storage vacuoles. Correction of storage, in both systems, resulted in the rapid decline of the mRNA to basal levels, with a slower decrease in the levels of lysosomal proteins. Lysosomal biogenesis in storage disorders is shown to be a regulated process which is partially controlled at, or prior to, the level of mRNA. Although lysosomal proteins were differentially regulated, the coordination of these events in lysosomal biogenesis would suggest that a common mechanism(s) may be in operation. PMID- 9223374 TI - The nuclear location of annexin V in the human osteosarcoma cell line MG-63 depends on serum factors and tyrosine kinase signaling pathways. AB - Serum starvation of MG-63 cells increases their doubling time from 24 h to 4 days. Cells grown in medium containing 10% fetal calf serum contain high levels of annexin V in the cell nucleus, whereas growth for 4 days in the absence of serum results in loss of nuclear annexin V from 72 +/- 4% of cells. Many of the cells which still have nuclear annexin V under these conditions seem to have recently finished dividing. Refeeding cells with medium containing serum restores annexin V to nuclei within 5 h. Charcoal treatment removes factors from serum that are required to allow annexin V to return to the nucleus. Protein synthesis is not required for annexin V to return to nuclei since inhibition of protein synthesis with cycloheximide does not prevent the serum-induced return of annexin V to nuclei. This, and other evidence, indicates that the presence of annexin V in nuclei reflects translocation rather than catabolism and resynthesis. Inhibition of tyrosine kinase activities with genistein attenuates the relocation of annexin V from the cytoplasm to the nucleus. Thus, the nuclear location of annexin V is controlled by signaling pathways involving serum factors and tyrosine kinases. The results argue for an important role for annexin V in the cell nucleus. PMID- 9223375 TI - Studies on rLMP7, a beta-subunit of the multicatalytic proteinase. AB - Subunit rLMP7 of the multicatalytic proteinase (MCP), which has been associated with chymotrypsin-like proteinase activity, was examined in rat liver and hepatocyte-derived cell lines. rLMP7 was detected in both nucleus and cytosol in liver by immunohistochemistry and immunoblotting, using a peptide-specific anti rLMP7 antibody. A M(r) 30,000 precursor protein was present only in cytosol, as was a minor component of M(r) 25,000. Mature rLMP7 (M(r) 23,000) was present in MCP in both nucleus and cytosol, although it was not detectable in the nuclear scaffold. Two rLMP7 cDNAs (designated rLMP7.1 and rLMP7.s) were identified by rapid amplification of 5' ends using RT/PCR, a result which was confirmed by Northern blot analysis and RNase protection assays. rLMP7.1 is 3-4x more abundant than rLMP7.s; it is 50 nt longer than the previously reported cDNA sequence and includes an upstream in-frame ATG within a consensus translation initiation sequence, which encodes the M(r) 30,000 rLMP7 precursor protein identified in vivo. rLMP7.s is 100 nt shorter than rLMP7.1 and does not contain the most 5' ATG. Transient transfection analyses with rLMP7.1 and rLMP7.s constructs coupled to green fluorescent protein showed that both transcripts were efficiently expressed in vivo. In vitro expression of these two rLMP7 cDNAs showed that rLMP7.1 produces the M(r) 30,000 precursor protein, whereas rLMP7.s produces two smaller peptides of M(r) 25,000 and 23,000. Purified 20S MCP preparations were able to proteolytically process the M(r) 30,000 precursor to the M(r) 25,000 product but not to the mature rLMP7 form. However, incorporation of this processed M(r) 25,000 product (or of either M(r) form produced from rLMP7.s) did not occur in vitro. In vitro processing and pulse-chase experiments suggested that the mature M(r) 23,000 subunit is derived, at least in part, from the M(r) 30,000 precursor. The M(r) 25,000 form may be a stable product produced directly from rLMP7.s. PMID- 9223376 TI - Time-resolved imaging of protein kinase C activation during sea urchin egg fertilization. AB - To study protein kinase C (PKC) activation during sea urchin egg fertilization we used three different fluorescent probes specific for PKC, namely, fim-1, which recognizes the catalytic site of the enzyme, and BODIPY- and NBD-phorbol esters interacting with the PKC regulatory domain. We were able to follow PKC activation during the early steps of fertilization, the three different probes giving the same fluorescent pattern. Within 120 s following insemination, the fluorescent signal increased and clustered in the cortical zone of the cell. The process was Ca2+ dependent and was inhibited in the presence of staurosporine, a PKC inhibitor. According to our in vitro probe characterization, this signal increase is due to PKC activation. These findings were further confirmed by Western blot analysis. This initial phase was followed by a rapid decrease which might be attributed to PKC hydrolysis by Ca2(+)-dependent proteases. The kinetics and the site distribution of PKC activation appear in complete agreement with the putative functions previously suggested for PKC during fertilization. PMID- 9223377 TI - 12-O-tetradecanoylphorbol-13-acetate inhibits aminophospholipid translocase activity and modifies the lateral motions of fluorescent phospholipid analogs in the plasma membrane of bovine aortic endothelial cells. AB - The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) is a potent mitogenic factor which can replace the growth promoting activity of basic fibroblast growth factor (bFGF) on bovine aortic endothelial cells. However, TPA treated cells lose their strict contact inhibition at confluence, which is a characteristic of cells grown in the presence of bFGF. We have examined whether these changes could be related to modifications of the transbilayer and lateral motions of fluorescent lipids, namely 1-acyl-2-[6-[N-(7-nitrobenz-2-oxa-1,3 diazol-4-yl)amino]caproyl]-p hosphatidylcholine (C6-NBD-PC), -phosphatidylserine (C6-NBD-PS), and -phosphatidylethanolamine (C6-NBD-PE) inserted in the outer leaflet of the cell plasma membrane. In TPA-treated cells, the three fluorescent phospholipids remained located in the outer leaflet for at least 1 h at 20 degrees C after their insertion, indicating a blockade of the aminophospholipid translocase activity which is normally present in the plasma membrane of bFGF treated cells. TPA also induced a large increase in the percentage of C6-NBD-PC and C6-NBD-PE probes which were free to diffuse laterally. The mobile fractions M reached values of approximately 100% for the two lipids, while for bFGF-treated cells they were found around 85 and 75%, respectively. For the C6-NBD-PS probe, M remained unchanged in bFGF and TPA-treated cells, at around 85%. TPA treatment also induced a twofold increase in the lateral diffusion coefficients of C6-NBD PC and C6-NBD-PE, while that of C6-NBD-PS remained nearly unchanged. These effects of TPA may be related to the observed loss of differentiated properties of vascular endothelial cells and not to its mitogenic properties. PMID- 9223378 TI - Uncoupling of S-phase and mitosis by recombinant cytotoxic necrotizing factor 2 (CNF2). AB - Cytotoxic necrotizing factor 2 (CNF2) is an exotoxin identified from virulent clinical isolates of Escherichia coli. It has been characterized in adherent cell lines as an inducer of cellular death, hyperploidy (multinucleation), and cytoskeletal reorganization. The molecular mechanism of these actions is unclear. Two cellular mechanisms can be hypothesized to explain the DNA content increase (hyperploidy) induced by the toxin. The first is that the toxin interferes with cytoplasmic division without interfering with normal nuclear cycling, such that DNA is replicated in the absence of cell division. The second is that the toxin drives the nuclear machinery to replicate the DNA multiple times within one cell cycle, without interfering with cytoplasmic division. In order to investigate these phenomena, we have constructed a recombinant CNF2 gene that expresses a toxin with both an epitope tag and a polyhistidine tag. Extracts made from E. coli that express this gene have a high multinucleating activity that colocalizes with the recombinant 115-kDa protein. To distinguish between these hypotheses, we used recombinant CNF2 and several growth conditions (time, partial differentiation, and stage of growth) to establish a relationship between cellular divisions and generation of hyperploidy. It was also determined that the toxin had no effect upon in vitro DNA replication using a Xenopus egg extract system. In aggregate, these data are consistent with the hypothesis that CNF2 is affecting cytoplasmic division and thereby removing the requirement for a completed mitosis before the initiation of another S-phase. These data are discussed in relation to the generation of polyploid cells during megakaryopoeisis and the generation of aneuploid cells during tumorigenesis. PMID- 9223379 TI - Fibroblast growth factor-1 induction of delayed-early mRNA expression in NIH 3T3 cells is prolonged by heparin addition. AB - Fibroblast growth factor (FGF)-1, also known as acidic FGF, is a multifunctional heparin-binding protein that is mitogenic for a wide variety of cell types cultured in vitro and a potent angiogenic agent in vivo. These cellular responses are mediated via high-affinity binding to a family of four membrane-spanning tyrosine kinase receptors. FGF-1-stimulated mitogenesis is potentiated by heparin, a sulfated glycosaminoglycan. In this study, we examined the effect of exogenous heparin on FGF-1-inducible gene expression in murine NIH 3T3 cells using both wild-type FGF-1 and FGF-1/glu132, an FGF-1 mutant with a reduced apparent affinity for heparin. The induction levels and temporal expression kinetics of two immediate-early response mRNAs (early growth response gene-1, thrombospondin-1) as well as two delayed-early response mRNAs (proliferin, ornithine decarboxylase) were monitored by Northern blot hybridization analysis. We found that although FGF-1 alone can promote the initial induction of these four mRNAs, heparin coaddition is necessary for prolonged delayed-early mRNA expression. This heparin effect occurs when cells are stimulated with wild-type FGF-1 but not with FGF-1/glu132. Furthermore, FGF-1 and heparin must be added together at the initial time of mitogen stimulation and they must remain present in the cell culture medium for a minimum period of 8 h to promote sustained delayed-early mRNA expression. These findings are consistent with the proposal that heparin promotes a long-term FGF-1:FGFR interaction which is required for sustained delayed-early gene expression and a full mitogenic response. PMID- 9223380 TI - Permeability and metabolic properties of a trophoblast cell line (HRP-1) derived from normal rat placenta. AB - The HRP-1 cell line is derived from normal rat placenta and appears morphologically similar to and retains characteristic expression of cellular markers of labyrinthine trophoblast cells. In this study, monolayers of HRP-1 cells grown on permeable supports were evaluated as a potential in vitro system to study trophoblast transport and metabolism. The cell line was shown to express and retain functional activity of the predominant placental cytochrome P450 isozyme, CYP1A1. Additionally, the HRP-1 cells retain functional activity of angiotensin I converting enzyme and carboxypeptidase N-like enzyme, peptidases characteristic of the trophoblast. The permeation of several hydrophilic, inert markers across the HRP-1 monolayers was observed to be dependent on effective molecular size and to be passive in nature. Functional asymmetry of the HRP-1 cells was illustrated by the predominant permeation of linoleic acid in the apical-to-basolateral direction across the monolayers. Transferrin passage across HRP-1 monolayers was concentration-dependent, was bidirectional, and could be inhibited by unlabeled transferrin, features typical of the trophoblast transport system for transferrin. Collectively, these properties suggest that the HRP-1 cell line may provide a useful tool for evaluating some of the permeability and metabolic properties of the trophoblast. PMID- 9223381 TI - alphaV Integrins on HT-29 colon carcinoma cells: adhesion to fibronectin is mediated solely by small amounts of alphaVbeta6, and alphaVbeta5 is codistributed with actin fibers. AB - HT-29 colon carcinoma cells form liver metastases upon intrasplenic injection, and adhesion to fibronectin under the liver microvascular liver endothelium is likely to be important for metastasis formation. We have therefore studied the integrins involved in fibronectin adhesion. This was not affected by blocking antibodies against the beta1, alpha3, and alpha5 integrin subunits, but it was blocked by an RGD-containing peptide, indicating involvement of RGD-dependent non beta1 alphaV integrins. Both alphaVbeta5 and alphaVbeta6 were detected on HT-29 cells. Blocking mAb against alphaV, but not against alphaVbeta5, abolished adhesion. From a HT-29 cell lysate, only alphaVbeta6 bound to a fibronectin Sepharose column. Thus, alphaVbeta6 is the main fibronectin receptor on HT-29 cells, despite the very low levels of alphaVbeta6 and the much higher levels of alphaVbeta5. The HT29 cells did not spread on fibronectin in the absence of serum, not even after a three- to fourfold increase in alphaVbeta6 levels, induced by interleukin 4. The cells did spread on vitronectin. Using immunofluorescence we observed that both on vitronectin and on fibronectin alphaVbeta5 was arranged in a striped pattern, aligned with actin fibers, and not in focal adhesions. On fibronectin, but not on vitronectin, alphaVbeta6 was concentrated in a punctate pattern at the periphery of cell islands. PMID- 9223382 TI - A paracrine role for myoepithelial cell-derived FGF2 in the normal human breast. AB - We have studied separated normal human breast epithelial and myoepithelial cells for the presence of basic fibroblast growth factor (FGF2) and its receptors, both low (heparan sulfate proteoglycans) and high affinity (FGFR1), and for the effects of FGF2 on the proliferation of both cell types. Our results indicate that these cells differ markedly in their synthesis and response to FGF2. We found, using PCR of purified cell populations, mRNA for FGF2 only in the myoepithelial cells, whereas immunostaining and Western blotting results demonstrated the presence of FGF2 protein in both epithelial and myoepithelial cells. FGF2 had no effect on the proliferation of myoepithelial cells, but it did maintain the survival of the separated epithelial cells in low serum and stimulate their growth in 5% and 10% FCS. Immunostainable FGFR1 was present in epithelial cells and, to a lesser extent, in myoepithelial cells. Low-affinity binding sites for FGF2 were synthesized by epithelial and myoepithelial cells, but myoepithelial cells possessed a greater proportion of higher-affinity heparan sulfate proteoglycans. These results indicate that myoepithelial cell-derived FGF2 may be an important paracrine factor controlling epithelial cell survival and growth in the normal human breast. PMID- 9223383 TI - Multiparametric staining to identify apoptotic human cells. AB - To analyze relevant features of HeLa and HL60 cells driven into apoptosis by etoposide, we have developed a new "tricolor" assay, based on the simultaneous analysis in the single cell of chromatin condensation, DNA degradation, and cellular poly(ADP-ribose) synthesis. The latter reaction is catalyzed by poly(ADP ribose)-polymerase (E.C. 2.4.2.30), an enzyme which is activated by the presence of DNA free ends. The protocol consists in the visualization of apoptotic cells by Hoechst staining, TUNEL assay, and immunoreaction with anti-poly(ADP-ribose) antibody. We thus provide the first evidence that endogenous poly(ADP-ribose) production is indeed stimulated in cells undergoing apoptosis after treatment with antitumoral drugs, and that the monitoring of this endogenous enzymatic reaction, combined with morphological and other biochemical parameters, should facilitate the detection of apoptotic cells. PMID- 9223384 TI - N6-isopentenyladenosine affects cAMP-dependent microfilament organization in FRTL 5 thyroid cells. AB - N6-Isopentenyladenosine (i6A), an adenosine and mevalonate derivative, inhibits, like adenosine, TSH-induced cAMP increase and its related events (I- uptake and DNA synthesis) in FRTL-5 cells. This inhibition is dose-dependent and is measurable at 10(-8) M. However, unlike adenosine, i6A prevents TSH-promoted microfilament disassembly. The effect of i6A on cytoskeletal structure is antagonized by pertussis toxin and could be assigned to its N6 substitution since it can be mimicked by other synthetic N6-adenosine derivatives. It is suggested that a step beyond cAMP is involved, since i6A prevents also microfilament disassembly induced by 8-bromo-cAMP. This is the first demonstration that an adenosine derivative, which is also an end-product of the isoprenoid pathway, affects cAMP-dependent microfilament organization. PMID- 9223385 TI - Centriole and centrosome dynamics during the embryonic cell cycles that follow the formation of the cellular blastoderm in Drosophila. AB - We have used immunofluorescence and electron microscopy to examine centrosome dynamics during the first postblastodermic mitoses in the Drosophila embryo. The centrosomal material, as recognized by antibodies against CP190 and gamma tubulin, does not show the typical shape changes observed in syncytial embryos, but remains compact throughout mitosis. Centrioles, however, behave as during the syncytial mitoses, with each daughter cell inheriting two separated centrioles at the end of telophase. During interphase in epithelial cells that have a distinct G1 phase, two isolated centrioles are found, suggesting that the separation of sister centrioles is tightly coupled to a mitotic oscillator in both the "abbreviated" and the "complete" embryonic division cycles. The centrioles of the Drosophila embryo sharply differed from the sperm basal body, having a cartwheel structure with nine microtubular doublets and a central tubule. This "immature" centriolar morphology was shown to persist throughout embryonic development, clearly demonstrating that these centrioles are able to replicate despite their apparently neotenic structure. PMID- 9223387 TI - The Lisbon Statement. World Health Organization (European Region) and the International Diabetes Federation (Europe). PMID- 9223386 TI - Statins and fibrates in the management of diabetic dyslipidemia. PMID- 9223388 TI - Community-based approaches for the primary prevention of non-insulin-dependent diabetes mellitus. AB - The prevalence of non-insulin-dependent (Type 2) diabetes mellitus (NIDDM) is increasing worldwide. Although recent studies suggest that primary prevention of NIDDM is possible, strategies for controlling the NIDDM pandemic remain under development. Successful interventions to date have mainly relied upon the control of obesity and increased exercise, although pharmacological agents are being studied. While a mixture of both high risk and population-based approaches is likely to be required, the former will not prevent new high risk cases developing. Unfortunately, the success in the primary prevention of cardiovascular disease through risk factor reduction has not controlled obesity, the most important risk factor for NIDDM. New strategies are currently being developed and focus upon changes in the food supply of whole populations and a community development approach to altering attitudes to food and exercise. As these interventions are in the early stages of their development, formative, process, and quantitative evaluation remain essential components of any community based programme aimed at the primary prevention of NIDDM. PMID- 9223389 TI - Measurement of markers of osteoclast and osteoblast activity in patients with acute and chronic diabetic Charcot neuroarthropathy. AB - Excess osteoclast activity is believed to be responsible for the early bone changes associated with Charcot neuroarthropathy in diabetes mellitus. Markers of osteoclast and osteoblast activity were measured in four groups of patients: 16 with an acute Charcot foot, 16 with a chronic Charcot foot, 10 diabetic controls, and 10 non-diabetic controls. Serum carboxyterminal telopeptide of type 1 collagen (1CTP), a marker of osteoclastic bone resorption, was significantly raised in the dorsal venous arch of the acute Charcot foot, 6.1 +/- 1.5 microg l( 1) (mean +/- SD) compared with the chronic Charcot foot 4.1 +/- 1.4, diabetic controls 3.3 +/- 1.4, and non-diabetic controls 2.8 +/- 1.4, p < 0.0001. This local increase in 1CTP was also reflected systemically in a study subgroup of 6 patients with acute Charcot neuroarthropathy, in whom peripheral antecubital vein 1CTP was 9.2 +/- 2.6 compared with 9.0 +/- 3.1 in the foot. In 6 chronic Charcot neuroarthropathy patients, foot (3.8 +/- 1.3) and systemic (4.0 +/- 1.5) 1CTP values were similar. Serum procollagen carboxyterminal propeptide (P1CP), an indicator of osteoblastic bone formation, was not significantly different between the feet of patients with acute Charcot neuroarthropathy 112 +/- 1.5 microg l( 1), patients with chronic Charcot neuroarthropathy 109 +/- 1.5 microg l(-1), diabetic controls 93.5 +/- 2.3 microg l(-1), and non-diabetic controls 90.1 +/- 1.5 microg l(-1). These results suggest that the acute Charcot foot demonstrates excess osteoclastic activity without concomitant increase in osteoblastic function. This may be important in its pathogenesis. PMID- 9223390 TI - Poor beta-cell function after the clinical manifestation of type 1 diabetes in children initially positive for islet cell specific autoantibodies. The Childhood Diabetes in Finland Study Group. AB - The prognostic significance of islet cell specific autoantibodies at the diagnosis of Type 1 (insulin-dependent) diabetes mellitus for the persistence of residual beta-cell function over the first 2 years of clinical disease was evaluated in a prospective population-based study. Seven hundred and eighty probands, aged 0.8-14.9 years, were examined for islet cell antibodies (ICA) and insulin autoantibodies (IAA), while 769 probands were studied for antibodies to glutamic acid decarboxylase (GAD65A). They were subsequently observed for 2 years. Lower serum C-peptide concentrations and higher requirement of exogenous insulin during the follow-up period were observed in the group of probands positive for at least one of the antibodies, especially for ICA or IAA. We conclude that the residual beta-cell function after the presentation of Type 1 diabetes is less in children initially positive for islet cell specific autoantibodies than in those testing negative at diagnosis. This might reflect possible heterogeneity in the pathogenesis of childhood diabetes. It also demonstrates that ICA and IAA negativity at the diagnosis of Type 1 diabetes is not associated with a smaller amount of functioning beta-cell mass, but the absence of antibodies probably reflects a slower beta-cell destructive process and a longer duration of preclinical disease. PMID- 9223391 TI - Left ventricular hypertrophy in non-insulin-dependent diabetic patients with and without diabetic nephropathy. AB - The aim of our cross-sectional case-control study was to evaluate putative mechanisms of the increased cardiac morbidity and mortality in NIDDM patients with or without diabetic nephropathy. Fifty-one NIDDM patients with diabetic nephropathy (38 males, age 61 +/- 8 years, group 1), 53 NIDDM patients with normoalbuminuria (42 males, 61 +/- 7 years, group 2), and 22 non-diabetic control subjects (15 males, 58 +/- 8 years, group 3) were investigated. Previous antihypertensive treatment was withdrawn 2 weeks before the study. Left ventricular mass index (LVMI) and systolic function were determined by echocardiography. LVMI was elevated, mean +/- SE, in group 1: 157 +/- 6 g m(-2), and in group 2: 139 +/- 7 g m(-2), as compared with group 3: 95 +/- 5 g m(-2) (p < 0.001, for both), and in group 1 as compared with group 2 (p = 0.05). The prevalence of left ventricular hypertrophy (LVH) (LVMI > 131 g m(-2) in men and > 100 gm(-2) in women) was much higher in group 1: 75% (95% CI, 60-86), and group 2: 51% (95% CI, 37-65), as compared with group 3: 9% (95% CI, 1-29) (p < 0.001, for both), and in group 1 as compared with group 2 (p < 0.01). Shortening fraction of the left ventricle, % +/- SE, was relatively reduced in group 1: 32.5 +/- 1.1%, and group 2: 33.4 +/- 1.1%, as compared with group 3: 41.2 +/- 1.2% (p < 0.01, for both). In a subgroup of 26 normoalbuminuric normotensive NIDDM patients, LVMI was higher than in 14 normotensive non-diabetic control subjects: 137 +/- 10 g m(-2) vs 96 +/- 7 g m(-2), respectively (p < 0.005). The prevalence of LVH was 42% (95% CI, 23-63) and 14% (95% CI, 2-43) (p = 0.07) in these two groups, respectively. In conclusion, normotensive and hypertensive NIDDM patients with and without diabetic nephropathy frequently suffer from LVH and relatively reduced systolic function which may constitute independent risk factors for fatal and non-fatal cardiac events. PMID- 9223392 TI - Pramlintide: a human amylin analogue reduced postprandial plasma glucose, insulin, and C-peptide concentrations in patients with type 2 diabetes. AB - In order to determine the influence of a 5 h infusion of pramlintide compared to placebo on postprandial glucose, lactate, insulin, and C-peptide concentrations in patients with Type 2 diabetes, a single-blind, randomized, cross-over study was conducted in 24 patients; 12 treated with exogenous insulin and 12 managed with diet and/or oral hypoglycaemic agents. One hour after initiation of infusion, patients consumed a Sustacal test meal. The protocol was repeated on the following day with each patient receiving the alternate study medication. Pramlintide infusion in the insulin-treated patients resulted in statistically significant reductions in mean glucose, insulin, C-peptide, and lactate concentrations during the 4-h period after the Sustacal test meal. Pramlintide infusion also resulted in significant reductions of mean insulin, C-peptide, and lactate concentrations, but not glucose concentrations, in the patients treated with diet and/or oral hypoglycaemic agents. Within this latter group, reduction in postprandial glucose concentrations in individual patients correlated with glycated haemoglobin values. These results suggest that administration of pramlintide may improve glycaemic control in patients with Type 2 diabetes treated with insulin or poorly controlled on diet and/or oral hypoglycaemic agents. PMID- 9223393 TI - Beta cell response to oral glimepiride administration during and following a hyperglycaemic clamp in NIDDM patients. AB - The aim of the present study was to assess the beta cell response to glimepiride, administered orally, during and following a hyperglycaemic clamp in 14 NIDDM patients (7 males), aged 62.5 (St. Dev. 7.7) years with a body mass index of 27.3 (2.8) kg m(-2) and HbA(Ic) of 7.0 (0.7)% at baseline, in a placebo controlled study. All patients were on stable treatment with a second generation sulphonylurea for at least 8 weeks prior to randomization and received placebo (P) or 5 mg glimepiride (G) daily for 7 days and 10 mg prior to a hyperglycaemic clamp (10.9 mmol l(-1) for 60 min, preceded by i.v. insulin infusion to stabilize fasting blood glucose levels at 4.0 mmol l(-1)). The clamp was followed by an observation period of 2 h in 5 subjects and 3.5 h in the next 9 subjects, during which blood glucose and plasma insulin, C-peptide and proinsulin levels were measured at regular intervals to determine the effect of glimepiride on the interaction between changes in glycaemia and plasma levels of beta cell products. Neither G nor P elicited a first phase insulin response. Areas under plasma insulin curve during the 1 h hyperglycaemic clamp were 94.2 (39.5) vs 69.1 (26.5) pmol.h l(-1) in G and P clamps, respectively (p = 0.002). Total areas (AUC) under the plasma insulin curve were 377 (145) vs 271 (113) pmol.h l(-1) in G and P clamps (< 0.05). Total AUCs of C-peptide were 309 (96) and 259 (102 pmol.h.(-1), in G and P clamps, respectively, p = 0.01. Total AUCs of proinsulin were 176 (77) versus 119 (56) pmol.h l(-1) in G and P clamps, respectively, p = 0.004. Five hours after G and P administration blood glucose levels were 4.7) 92.1) mmol(-1) in the G clamp vs 6.2 (1.9) mmol l(-1) in the P clamp (p = 0.001). The number of hypoglycaemic events (blood glucose < 3.0 mmol l(-1)) in the 3.5 h observation period was 3 in G clamps vs 0 in P clamps (p = ns). In conclusion, glimepiride stimulates the second phase insulin and proinsulin secretion. The lowering of blood glucose levels is not accompanied by a commensurate inhibition of the insulin secretion. Further studies are required to compare this new drug with currently available oral hypoglycaemic agents, with respect to glycaemic control and the risk of hypoglycaemia. PMID- 9223394 TI - Comparison of bezafibrate and simvastatin in the treatment of dyslipidaemia in patients with NIDDM. AB - Fibrates and HMG CoA reductase inhibitors are commonly used in the treatment of diabetic dyslipidaemia. However, these two groups of drugs have not been compared in diabetic patients in a randomized controlled trial. Therefore, a multicentre study was performed in 73 subjects with non-insulin-dependent (Type 2) diabetes mellitus (NIDDM) and combined hyperlipidaemia (serum cholesterol 6.2-10.0 mmol l( 1), serum triglycerides 2.3-10.0 mmol l(-1)), comparing the efficacy of 400 mg bezafibrate with 10 mg simvastatin in a double-blind fashion. Treatment with bezafibrate during 12 weeks reduced serum triglycerides significantly more than simvastatin (-41% vs -22%, p < 0.001) and increased HDL cholesterol more (bezafibrate: + 17% vs simvastatin: + 9%, p < 0.05). LDL cholesterol levels decreased by 14% (p < 0.001) during simvastatin and increased by 21% (p < 0.01) during bezafibrate. This increase in LDL cholesterol was positively correlated with fasting serum triglycerides (p < 0.001) and was associated with a reduction of the serum apolipoprotein B concentration, suggesting an increase in LDL particle size. Metabolic control of diabetes (fasting glycaemia; HbA1c) and insulin secretion (C-peptide levels) were unaffected by both treatments. The incidence of side-effects during treatment was similar for both drugs. Thus, 400 mg bezafibrate mainly increases HDL cholesterol and lowers serum triglycerides but at the expense of an increase in LDL cholesterol; 10 mg simvastatin lowers LDL cholesterin more effectively but has a smaller effect on HDL cholesterol and triglycerides. PMID- 9223395 TI - Blindness due to diabetes: population-based age- and sex-specific incidence rates. AB - Reducing the incidence of diabetic retinopathy and blindness was declared one of the main objectives in St Vincent. To date, hardly any valid data are available on the age- and sex-specific incidence of diabetes-related blindness. They are necessary, however, to evaluate intervention activities. Therefore, we used a population-based registry of blindness to calculate incidence of blindness due to diabetes. In one German district (Rhineland) we obtained complete lists of cases of blindness newly registered in 1990 and 1991 and coded as blind due to diabetes (n = 589). We estimated age-specific incidence rates in the entire as well as in the diabetic population. Incidence rates of blindness due to diabetes (100,000( 1) * year(-1)), standardized to the West-German population, were 3.2 (CI 95%: 2.9;3.4) in the entire population and 60.5 (CI 95%: 45.7;75.4) in the diabetic population. Incidence rates in the diabetic population showed a peak between 20 and 40 years of age, probably due to complications of Type 1 diabetes. Incidence was higher in diabetic women than in diabetic men (p < 0.05 at ages > or =40 years). Repeating the study will detect a decrease in the incidence of blindness due to diabetes by one-third with over 99% power. PMID- 9223396 TI - Mortality in diabetic patients participating in an ophthalmological control and screening programme. AB - The aim of this follow-up study has been to assess retinopathy and change of treatment to insulin therapy as risk factors for mortality in diabetic patients participating in a control and screening programme for retinopathy. A total of 3220 diabetic patients, 483 with an age at diagnosis <30 years, and 2737 with an age at diagnosis > or = 30 years, were included. Retinopathy was graded on fundus photographs using the Wisconsin Scale, and the visual acuity was assessed. The average HbA1c value was calculated for each patient for the previous 8 years to estimate long-term glycaemic control. Mortality data were obtained from death certificates. Two hundred and sixty-three diabetic patients (8.2%) died during the mean follow-up time of 3.4 years, 13 (2.7%) of those with younger-onset (<30 years) and 250 (9.1%) of those with older-onset (> or = 30 years) diabetes. Of them, 148 (56.3%) died from cardiovascular and 23 (8.7%) from cerebrovascular disorders. After adjusting for differences in age and sex, more severe retinopathy and the use of antihypertensive drugs were associated with a decreased overall survival rate as well as an increased mortality from cardiovascular and cerebrovascular diseases. A statistically significant association between HbA1c values in the highest quartile, i.e. > or =8.4%, and cardiovascular and all cause mortality did not remain when retinopathy was entered into the multivariate analyses. Duration of diabetes, but not change of treatment to insulin therapy, was associated with higher cardiovascular mortality in patients whose diabetes was diagnosed after the age of 30 years. We conclude that severe retinopathy, use of antihypertensive drugs, and poor glycaemic control predicted death from cardiovascular disease in diabetic patients participating in an ophthalmological screening programme. PMID- 9223397 TI - Standardization of glycated haemoglobin assays throughout the Northern region of England: a pilot study. AB - We describe a pilot study designed to assess the comparability of measurements of glycated haemoglobin among 15 laboratories in the North of England. We also evaluated a means of improving agreement and aligning results by referencing locally measured values to those obtained by the central biochemistry laboratory of the Diabetes Control and Complications Trial research group. Blood samples from 50 diabetic and non-diabetic subjects were analysed for glycated haemoglobin content in the participating and reference laboratories using a variety of routinely available methods. The mean CV for these results was 15.3% (95% confidence interval 14.0% to 16.5%). Using a regression equation relating a subset of seven of these results to their assigned reference values, a glycated haemoglobin index was calculated for all the other samples distributed. The mean inter-laboratory CV improved to 4.6% (95% confidence interval 4.0% to 5.1%), p<0.0001. The percentage bias of results from the reference method also improved from 15.1% (95% confidence interval 9.4 to 20.1) to 4.67% (95% confidence interval 4.05 to 5.25) after alignment, p<0.001. This study demonstrated that substantial method related differences between reported glycated haemoglobin results exist. These can be reduced using a simple calibration strategy in which data are correlated to an established method with associated extensive clinical interpretive value as established by the DCCT. PMID- 9223398 TI - Hospital utilization as a function of social deprivation: diabetes vs non diabetes. AB - We tested the hypothesis that a relationship between ill health and deprivation exists for patients with diabetes, distinct from that experienced by the non diabetic population. Age standardized admission and appointment rates and proportion of total activity for patients with and without diabetes were determined by electoral ward and correlated with the Townsend index of social deprivation for the health district of South Glamorgan (population 408,000). Both diabetic (r = 0.78, p < 0.001) and non-diabetic (r = 0.74, p < 0.001) in-patient admissions were positively correlated with social deprivation. This relationship also existed for attended out-patient appointments (r = 0.67, p < 0.001 and r = 0.45, p < 0.01, respectively). The proportion of diabetic to non-diabetic admissions by ward also showed a positive correlation for in-patients (r = 0.47, p < 0.001). This remained true for IDDM (r = 0.23, not significant) and NIDDM (r = 0.62, p < 0.001) diabetes, for admissions for coronary heart disease (r = 0.50, p < 0.001) and cerebrovascular disease (r = 0.29, p < 0.05), elective admissions (r = 0.30, p > 0.05), and emergency admissions (0.46, p < 0.001). Our results suggest that secondary care utilization is positively correlated with social deprivation and that this relationship is stronger in the diabetic population. This may be due to different prevalence rates or increased complications requiring hospital treatment in different social circumstances. Further research is required to examine these factors more closely. PMID- 9223399 TI - Prevalence of glucose intolerance in urban and rural communities in Saudi Arabia. AB - The prevalence and associated factors for glucose intolerance among Saudi populations in urban and rural communities were investigated among 13177 subjects, 15 years and over, from different regions of Saudi Arabia. The data were standardized using the known age structure of the Saudi population. The sample was randomly selected from subjects who participated in the National Epidemiological Household Study for Chronic Metabolic Diseases. Medical and social history was ascertained from all the study population during house visits. All subjects were then requested to attend a local primary care centre for physical examination and phlebotomy for measurement of random plasma glucose (RPG). A 75 g oral glucose tolerance test was employed for subjects with borderline values. WHO criteria for diagnosis of diabetes mellitus (DM) and impaired glucose tolerance (IGT) were applied. Mean RPG from the urban population was significantly higher than in the rural population. Age adjusted prevalence of DM was significantly higher in the urban population (males 12%, 95% CI 11-13 and females 14%, 95% CI 13-15) than in the rural population (males 7%, 95% CI 7-8 and females 7.7%, 95% CI 7-9) and is among the highest in the world. The prevalence of DM increased with age. The lowest and highest prevalences of DM in the urban population were 2% for subjects aged 15-20 years and 49% for female subjects aged 51-60 years. The lowest and highest prevalences of DM among rural population were 1% for subjects aged 15-20 years and 29% for female subjects over the age of 60 years. Fifty-six per cent of diabetic patients were newly diagnosed at the time of the study. Age adjusted prevalence of IGT was not significantly higher in the urban population. The highest prevalence of obesity, BMI>30, was among urban female subjects. Age, obesity, and family history of DM were associated with DM. Considering the young nature of Saudi population, the prevalence is expected to increase in the near future. There is a need to develop a multi-disciplinary approach for the general population with special attention to female subjects for prevention through controlling modifiable risk factors such as obesity and sedentary life style, improving glycemic control of the diabetic population, and early identification and treatment of diabetic complications. PMID- 9223400 TI - Clinical characteristics and outcome of hyperglycaemic emergencies in Johannesburg Africans. AB - In this prospective analysis we investigated the clinical characteristics of black South African diabetic patients admitted to hospital with hyperglycaemic emergencies. The study cases were selected from the medical admissions to an urbanized, Johannesburg academic hospital over a period of 12 months. Only patients with severe diabetic ketoacidosis (DKA) or hyperosmolar non-ketotic hyperglycaemia (HNKH) as defined in the text were included. Over the study period, we identified 58 patients with severe DKA (M: 32, F: 26) and 24 with HNKH (M: 14, F:10). Thirty-two of the patients with DKA (55.2%) were classified as having non-insulin dependent (Type 2) diabetes mellitus (NIDDM). Compared to the 26 subjects with insulin-dependent (Type 1) diabetes mellitus (IDDM), the NIDDM patients were older (51.7 vs 27.7 years) and had a significantly higher body mass index (BMI) (29.4 vs 23.5 kg m(-2), p = 0.002), and glucose levels 47.5 vs 34 mmol l(-1) p = 0.004). Mortality from DKA was 6.8 % and from HNKH 16.6%. Infection was the leading precipitating factor for both DKA and HNKH, followed by first presentation and noncompliance. We conclude that the majority of urban African patients admitted to hospital with DKA have NIDDM. Mortality from DKA among the black Africans in Johannesburg is low and comparable to the mortality in western Europe. PMID- 9223401 TI - Fatal hand sepsis in Tanzanian diabetic patients. AB - Hand infections are common presentations among diabetic patients admitted to hospital in Tanzania. The morbidity and mortality are high and patients' hospital inpatient stay tend to be prolonged because of suboptimal therapy. We describe four diabetic patients with hand infections and fatal outcomes. In contrast to patients with foot infections, none of our patients had clinical evidence of peripheral neuropathy or vascular disease. All four patients eventually died in hospital after acquiring hand sepsis and diabetic ketoacidosis which did not respond to prolonged courses of intravenous insulin and antimicrobials. Literature review suggests such infections are at least as likely to include Gram negative organisms as Staphylococcus aureus. Primary management should have included aggressive surgery with limb amputation if necessary with adjunctive antimicrobial therapy. PMID- 9223402 TI - A strategy for arterial risk assessment and management in type 2 (non-insulin dependent) diabetes mellitus. European Arterial Risk Policy Group on behalf of the International Diabetes Federation European Region. AB - People with Type 2 (non-insulin-dependent) diabetes mellitus die mainly from cardiovascular and cerebrovascular disease. Furthermore, the major burden of their symptoms arise from arterial disease, including peripheral vascular disease. However, management guidelines for Type 2 diabetes continue to focus on blood glucose control, which is only one of a number of arterial risk factors found with this type of diabetes. Clinically it is evident that blood glucose control continues to occupy centre-stage in the management of Type 2 diabetes as practised by many physicians. Even when arterial risk factors such as smoking or raised serum triglycerides are noted, their management is often relatively neglected. As part of the St Vincent Declaration Action Programme, a working group has sought consensus on the number and relative importance of arterial risk factors requiring management in quality diabetes care. The group seeks to assist those devising protocols and guidelines, records and quality systems, and those charged with directly advising and educating people with diabetes. Arterial risk factors that can be routinely identified and monitored, and modified by application of management protocols, include high blood pressure, high serum total and LDL cholesterol, low serum HDL cholesterol and raised serum triglycerides, poor blood glucose control, smoking, high body mass index and body fat distribution. Aspirin can modify hypercoagulability, but this is not easily monitored. Arterial risk factors that cannot be modified, but which have an impact on the intensity of management of other factors, include ethnic group, gender, and family history of arterial disease. Raised albumin excretion is an arterial risk factor and can be modified, but it is not clear whether this reduces cardiovascular risk. For many of the risk factors, levels of high, medium, and low risk can be set. These can be used, in consultation with the patient, to determine appropriate interventions and provide feedback on risk reduction resulting from successful management. PMID- 9223403 TI - Alternatives to the retinal camera. PMID- 9223404 TI - Dietetic advice in general practice. The Nutrition Sub-Committee of the British Diabetic Association. PMID- 9223405 TI - Scientific, logistical, and ethical issues related to creation of a stem cell repository for use in experimental trials of gene therapy for AIDS. PMID- 9223406 TI - Resistance to immunodeficiency viruses and its relevance to vaccine development. PMID- 9223407 TI - Protection of SIVmac-infected macaque monkeys against superinfection by a simian immunodeficiency virus expressing envelope glycoproteins of HIV type 1. AB - The infection of macaque monkeys by attenuated simian immunodeficiency virus can vaccinate against pathogenic molecular clones and isolates of the same virus. The correlates of this potent protective immunity are not fully understood but may be the key to an effective AIDS vaccine for humans. Aiming to determine whether host immune responses to envelope glycoprotein are an essential component of the immunity to primate lentiviruses, we have tried to superinfect SIVmac-infected macaque monkeys with SHIVsbg, a chimeric primate lentivirus constructed from the SIVmac239 genome with the env, rev, tat, and vpu genes from HIV-1 Lai. After inoculation of a large dose of SHIVsbg, the chimeric virus was isolated by coculture of mononuclear blood cells from four of five SIV-infected monkeys, but three animals were protected from extracellular SHIV viremia and did not seroconvert to HIV-1 glycoproteins. In the two SIV-infected monkeys that did develop SHIV viremia, cell-associated viral load was reduced at least 100-fold. These data indicate that an antiviral response capable of effectively controlling primate lentivirus replication might not necessarily involve the envelope glycoprotein. PMID- 9223408 TI - Anti-major histocompatibility complex antibody responses to simian B cells do not protect macaques against SIVmac infection. AB - Macaques have been protected against infection with human cell-grown SIVmac by immunization with antigens encoded by the human major histocompatibility complex (MHC). Here, we investigated the efficacy of alloimmunization with simian B cells expressing high levels of MHC class I and class II molecules to confer protection against systemic challenge with simian-grown SIVmac. Eight rhesus macaques were vaccinated with glutaraldehyde-fixed and beta-propiolactone-inactivated herpesvirus papio-transformed B cells. Four of the macaques received 5 doses, the others 10. Animals were challenged with rhesus macaque spleen-derived cell-free SIVmac. Allogeneic B cells elicited antibody responses to rhesus MHC class I and II but failed to protect animals against infection. Anti-MHC class I antibodies were restricted in specificity and failed to recognize MHC class I from some B lymphoblastoid cell lines (B-LCLs) including a B-LCL from the animal in whose cells the challenge virus was grown. Vaccinated animals responded to self-MHC class I antigens but not to self-MHC class II antigens from peripheral blood mononuclear cells (PBMCs). Animals that underwent the shorter immunization regimen had transiently enhanced PBMC-associated virus loads after challenge, whereas the average virus-infected cell load was reduced in animals that underwent the more extensive immunization. These results suggest that antibody responses to allogeneic MHC molecules do not protect against infection with immunodeficiency lentiviruses. PMID- 9223410 TI - Intranasal immunization is superior to vaginal, gastric, or rectal immunization for the induction of systemic and mucosal anti-HIV antibody responses. AB - Vaginal anti-HIV antibody responses may be beneficial, and possibly required, for vaccine-induced protection against HIV infection acquired through receptive vaginal intercourse. We have previously determined that intranasal immunization with a hybrid HIV peptide and cholera toxin induced vaginal anti-HIV IgA responses in BALB/c and C57BL/6 mice. To determine if vaginal, gastric, or rectal boosting would enhance the induction of vaginal anti-HIV IgA responses over those observed with intranasal immunization only, C57BL/6 mice were intranasally immunized with the hybrid HIV peptide T1SP10MN(A) and cholera toxin (days 0 and 14) and boosted via the vaginal, gastric, or rectal route (days 7 and 28). Four intranasal immunizations was superior to all other immunizations evaluated for the induction of plasma anti-peptide IgG, vaginal anti-peptide IgG and IgA, and peptide-specific delayed-type hypersensitivity. In addition, intranasal priming with gastric boosting was associated with greatly elevated total serum IgE concentrations whereas intranasal immunization only was associated with only a modest increase in total serum IgE. These results suggest that intranasal immunization is a viable route of immunization for the induction of systemic and mucosal anti-HIV immune responses. PMID- 9223409 TI - A humanized form of a CD4-specific monoclonal antibody exhibits decreased antigenicity and prolonged plasma half-life in rhesus monkeys while retaining its unique biological and antiviral properties. AB - Certain monoclonal antibodies (MAbs) directed against CD4 can efficiently block HIV-1 replication in vitro. To explore CD4-directed passive immunotherapy for prevention or treatment of AIDS virus infection, we previously examined the biological activity of a nondepleting CD4-specific murine MAb, mu5A8. This MAb, specific for domain 2 of CD4, blocks HIV-1 replication at a post-gp120-CD4 binding step. When administered to normal rhesus monkeys, all CD4+ target cells were coated with antibody, yet no cell clearance or measurable immunosuppression occurred. However, strong anti-mouse Ig responses rapidly developed in all monkeys. In the present study, we report a successfully humanized form of mu5A8 (hu5A8) that retains binding to both human and monkey CD4 and anti-AIDS virus activity. When administered intravenously to normal rhesus monkeys, hu5A8 bound to all target CD4+ cells without depletion and showed a significantly longer plasma half-life than mu5A8. Nevertheless, an anti-hu5A8 response directed predominantly against V region determinants did eventually appear within 2 to 4 weeks in most animals. However, when hu5A8 was administered to rhesus monkeys chronically infected with the simian immunodeficiency virus of macaques, anti hu5A8 antibodies were not detected. Repeated administration of hu5A8 in these animals resulted in sustained plasma levels and CD4+ cell coating with humanized antibody for 6 weeks. These studies demonstrate the feasibility of chronic administration of CD4-specific MAb as a potential means of treating or preventing HIV-1 infection. PMID- 9223412 TI - Lack of interleukin 10 expression in monocyte-derived macrophages in response to in vitro infection by HIV type 1 isolates. AB - Interleukin 10 is a pleiotropic cytokine that is overexpressed in HIV-infected patients. Here, we investigated IL-10 expression in primary cultures of monocyte derived macrophages (MDMs) in response to in vitro infection by HIV-1/Ba-L or two macrophage-tropic HIV-1 primary isolates. Whatever the multiplicity of infection used, and in spite of high replication levels and an increase in HIV-infected cell frequency, neither significant IL-10 secretion nor IL-10 mRNA overexpression was induced in HIV-1-infected MDMs. Moreover, identical results were obtained with HIV-1-infected 1-day monocytes. These results show that MDM infection by HIV is not sufficient by itself for inducing IL-10 synthesis. PMID- 9223411 TI - Cytotoxic monocytes in the blood of HIV type 1-infected subjects destroy targeted T cells in a CD95-dependent fashion. AB - HIV-1 infection changes the functional balance of macrophages in the body; it inhibits the development of macrophages capable of costimulating T cell responses and it favors the development of macrophages that kill T cells with which they form cellular conjugates. Cytotoxic macrophages destroy CD4 T cells, which they target through CD4-reactive immune-complexed HIV-1 envelope molecules on a large scale. They also destroy T cells that they target through presented antigen or mitogen. We show here that cytotoxic macrophages destroy their cellular targets at least partially in a CD95-dependent process in which T cells first modulate expression of most of their membrane receptors and subsequently die. PMID- 9223413 TI - Preliminary findings of an in vitro human spleen mononuclear cell culture system for primary isolates of HIV type 1. AB - Acute HIV-1 infection is often manifested with a high level of viremia. The cell types and tissues/organs that contribute to the virus load are thought to be of central and peripheral lymphoreticular origin. The establishment and permissiveness of organ-based cell culture systems from spleen with laboratory strains or primary isolates of HIV-1 have not been reported. We studied unseparated splenic mononuclear cells (SMCs) and adherent cells derived from human spleen and liver in comparison with blood monocyte-derived macrophages (MDMs). Unstimulated, SMCs were highly permissive to primary lymphotropic HIV-1 and dual/macrophage-tropic isolates (which are able to replicate in both MDMs and PBMCs). Furthermore, SMCs were found to replicate virus to high titer in a rapid log-phase manner and exhibited a prolonged stationary phase of virus production, unlike PBMCs, which required conventional activation with mitogens and exhibited a much shorter period of virus production. Interestingly, the SMCs maintained themselves as a mixed phenotype of nested lymphocytes with complex and well differentiated macrophage(s) for extended periods of time. In addition, splenic macrophages readily purified by adherence were highly permissive to a dual/macrophage-tropic primary isolate, HIV-1ADA, intermediate with two laboratory strains, HIVR-1RF and HIV-lHXB3, and least permissive to the lymphotropic primary isolate HIV-1Mr452 and two other laboratory strains, HIV-1CC and HIV-1MN. The replication of HIV-1ADA as measured by extracellular p24 was sustained for up to 7 weeks and similar to the replication patterns observed with adherent hepatic macrophages and blood-derived MDMs. This study demonstrates that exogenous stimulation is not required for infection of these cells; either adherence-isolated and/or mixed lymphoid populations can be studied together, and viable stocks can be readily prepared and cryopreserved. In addition, these cells could be used for isolating new and/or other variants of HIV-1. Thus, the use of the SMC primary in vitro cell culture system for future studies involving HIV-1 is warranted. PMID- 9223414 TI - Phenotypic switch in a Spanish HIV type 1 isolate on serial passage on MT-4 cells. AB - A biological clone (F0) of a syncytium-inducing (SI) isolate (S61) was unable to produce syncytia in MT-4 cells. On serial passage on MT-4 cells this virus [F15( 3)] became capable of inducing syncytia (Sanchez-Palomino S, et al.: J Virol 1993;67:2938). After sequencing different regions of the env gene including V1 V2, V3, and the fusion domain of both viruses, we have found only an asparagine (N)-to-isoleucine (I) change in position 7 of the V3 loop. By mutagenesis and in vitro recombination, using infectious molecular clones, we have identified this amino acid change as the only one responsible for the syncytial phenotypic switch. However, this cytopathic change was not accompanied by a change in the replication rate, indicating that these two properties are not linked genotypic traits. Thus serial passaging of an HIV-1 isolate on MT-4 cells has produced a nonsyncytial-to-syncytial switch through a point mutation in position 7 of the V3 loop. PMID- 9223415 TI - The shape of cell death. AB - Cell death, a scheduled event during development and tissue turnover, or the ultimate consequence of toxic or pathologic insults seems to involve a relatively limited number of execution pathways. This reflects the evolution of an organized sequence of events perhaps converging onto final common pathways that are used to dispose of unwanted or injured cells. In many cases, the ordered execution of this internal death program leads to typical morphological and biochemical changes that have been termed apoptosis. Apoptosis, often equated with developmental or programmed cell death, has been opposed to unscheduled or accidental cell lysis/necrosis. However, increasing evidence suggests that the two forms of cell demise share similar characteristics, at least in the signaling and early progression phase. Recent studies have shown that, when the intensity of the insult is very high and/or when ATP generation is deficient, cells fail to execute the ordered changes ensuing in apoptosis. Then cell lysis/necrosis supervenes before the processes leading to nuclear condensation and exposure of surface molecules can be completed. Thus, apoptosis and necrosis seem to represent only different shapes of cell demise, resulting from a more or less complete execution of the internal death program. PMID- 9223417 TI - Limited proteolysis of human kidney angiotensin-converting enzyme and generation of catalytically active N- and C-terminal domains. AB - The somatic form of angiotensin converting enzyme is a class I ectoenzyme that is bound to the surface of endothelial calls. It consists of two homologous, catalytic domains of approximately 600 residues each; a juxtamembrane "stalk" region; a transmembrane, hydrophobic sequence; and a 30 residue, C-terminal cytosolic domain. We have used limited proteolysis to probe the structural and functional properties of the enzyme. Endoproteinase Asp-N cleaves both the Thr615 Asp616 and the Leu1219-Asp1220 peptide bonds to generate the two catalytic domains which were isolated by a combination of immunoaffinity and lisinopril Sepharose affinity chromatography. The enzymatic characteristics of the N and C fragments were examined with angiotensin I, hippuryl-His-Leu, and luteinizing hormone-releasing hormone and indicate that both fragments contain catalytically active sites that retain their individual functional integrity. PMID- 9223416 TI - Purification, cloning, and expression of human plasma hyaluronidase. AB - Hyaluronidase was purified from human plasma using Triton X-114 phase extractions and ion-exchange chromatography. Monoclonal antibodies generated against the purified protein by a novel screening assay were utilized to isolate homogeneous enzyme for microsequencing. The amino acid sequences obtained matched a cDNA in the Expressed Sequence Tag database which, with 5'-RACE-PCR, was used to clone the plasma hyaluronidase gene, termed Hyal-1. Hyal-1 codes for a protein of 435 amino acids that is over 40% identical to PH-20, a sperm-specific hyaluronidase. Unlike PH-20, which is only expressed in testis, transcripts of Hyal-1 were found in multiple tissues. Hyal-1 stably expressed in human embryonic kidney cells resulted in a 3,000 fold increase of secreted immunoreactive hyaluronidase activity that was biochemically indistinguishable from human plasma hyaluronidase. By immunological, molecular and biochemical criteria, we conclude that Hyal-1 is the predominant hyaluronidase found in human plasma. PMID- 9223418 TI - Detection of mouse skeletal muscle-specific product, which includes ZF5 zinc fingers and a VP16 acidic domain, by reverse transcriptase PCR. AB - ZF5, which we have cloned as a repressor on the mouse c-myc promoter, is a zinc finger protein containing Kruppel-type zinc finger and ZiN/POZ domains. In a reverse transcriptase PCR assay using mouse skeletal muscle RNA, we identified a 827 bp PCR product including the zinc finger domain of ZF5 and the acidic domain of VP16. The presence of the VP16 acidic domain induced the reduction of DNA binding activity of the zinc finger domain. In addition, the inhibitory effect of the VP16 acidic domain was demonstrated on the human immunodeficiency virus (HIV) promoter, but there was no effect on the thymidine kinase (TK) promoter. PMID- 9223419 TI - Metabolism of glycerol-1,2,3-trimethylsuccinate in rat pancreatic islets. AB - The metabolism of 14C-labelled glycerol-1,2,3-trimethylsuccinate (2.0 mM) was examined in rat pancreatic islets. The oxidation of the glycerol moiety of the ester was negligible relative to that of its succinate residues. The oxidation of glycerol-1,2,3-trimethyl[1,4-(14)C]succinate was two times higher than that of glycerol-1,2,3-trimethyl[2,3-(14)C]succinate, this difference being matched by a higher generation of 14C-labelled acidic metabolites and amino acids from the latter than from the former tracer. The total generation of 14CO2 from the ester, uniformly labelled except in its methyl groups, was close to that found for the oxidation of 1.0 mM D-[U-(14)C]glucose. These findings thus reveal that glycerol 1,2,3-trimethylsuccinate is efficiently metabolized in islet cells and support the idea that this ester could be used as a nutrient to bypass defects of D glucose transport and metabolism in the islet B-cell and, hence, improve proinsulin biosynthesis and insulin release in non-insulin-dependent diabetes mellitus. PMID- 9223420 TI - Chromatin neutral sphingomyelinase and its role in hepatic regeneration. AB - Ceramide acts as a second messenger and modulates many cellular functions. This molecule can be produced by enzymatic digestion of sphingomyelin, a phospholipid which has been shown to be in high concentration in a chromatin phospholipidic fraction. In order to clarify whether chromatin sphingomyelin has a role in this signal transduction pathway, it is necessary to define the sphingomyelin cycle. Neutral sphingomyelinase represents the first step of the cycle. In this paper we demonstrate that sphingomyelinase activity can be detected also in the chromatin of rat hepatocyte nuclei and it increases in relation to the entrance in S phase of hepatocyte after hepatectomy. The enzyme can be distinguished from that present in the nuclear envelope on the basis of optimum pH and Km. The role of the spingomyelin pathway in relation to liver regeneration is discussed. PMID- 9223421 TI - Perforin-enhancing protein, a low molecular weight protein of cytotoxic lymphocyte granules, enhances perforin lysis. AB - Perforin is a 68 kD protein found in the granules of cytotoxic lymphocytes and is used by lymphocytes to form lethal pores in the membranes of the cells they kill. We and others have found that when perforin is purified, its lytic activity is markedly reduced. ELISAs indicated that our final recovery of perforin protein was excellent. We decided to determine if depletion of other granule proteins contributed to the loss of lytic activity. We isolated perforin to the point where lytic activity was diminished and added back granule proteins that had no lytic activity or detectable (antigenic) perforin. Perforin was isolated by Cu2+ immobilized metal affinity chromatography (IMAC) followed by phenyl-Superose hydrophobic interaction chromatography (HIC). Its lytic activity was enhanced by a low molecular weight (<15 kD) protein, perforin enhancing protein (PEPr). We have isolated PEPr by two methods, HIC and MonoQ. Nonlytic PEPr restored perforin to close to its original lytic activity. A protein similar if not identical to PEPr was also detectable as an 125I-labeled protein associated with lytic perforin. We propose that PEPr acts in conjunction with perforin to form lethal pores and suggest that PEPr may be the rat equivalent of the human cytotoxic lymphocyte protein, granulysin. PMID- 9223422 TI - The proteasome is an essential mediator of the activation of pre-MPF during starfish oocyte maturation. AB - Starfish oocyte maturation was blocked by the addition of 100 microM MG115, a potent proteasome inhibitor, whereas no inhibition was observed by membrane permeable cysteine protease inhibitor, E-64-d. The inhibition by MG115 was diminished by adding at a time corresponding to the half time required for germinal vesicle breakdown. Potent inhibition of germinal vesicle breakdown was also observed by microinjection of anti-proteasome-a-subunit antibodies. The antibody-injected oocytes failed to activate pre-maturation promoting factor (pre MPF), since the dephosphorylation of phospho-Tyr15 in cdc2 kinase was not observed even in the presence of 1-methyadenine, a maturation-inducing hormone. These results indicate that the proteasome triggers the activation of pre-MPF via the dephosphorylation of cdc2 kinase in the signal transduction pathway in response to the hormonal stimulus during starfish oocyte maturation. PMID- 9223423 TI - The complete coding sequence of hepatitis C virus genotype 5a, the predominant genotype in South Africa. AB - Hepatitis C virus (HCV) genotype 5a is the predominant genotype in southern Africa with a high prevalence amongst infected blood donors from areas in South Africa. We have determined the nucleotide sequence corresponding to the complete coding region of an HCV isolate, EUH1480, previously classified as genotype 5a, from an Edinburgh haemophiliac. The sequence contained a single open reading frame (ORF) coding for a polyprotein of 3014 amino acids. Comparison with the polyprotein sequences from other HCV genotypes, where the ORF varies from 3008 to 3037 amino acids, showed the observed variation in size was due to differences in lengths of the envelope 2 and the nonstructural 5A proteins. The sequence divergence of HCV genotype 5 ranged from 29.4% nucleotide differences (24.91% amino acid differences) compared with genotype 1c to 32.5% nucleotide differences (30.3% amino acid differences) compared with 2a. Phylogenetic analysis of the available full length nucleotide sequences showed EUH1480 to form a branch distinct from the other HCV types, confirming the classification of type 5a as a separate genotype. PMID- 9223424 TI - Sac1p of Saccharomyces cerevisiae is not involved in ATP release to the extracellular fluid. AB - One activity ascribed to Sac1p is the transport of ATP into the lumen of the endoplasmic reticulum of Saccharomyces cerevisiae; therefore, the question of whether this protein plays a role in ATP efflux from yeast was addressed. Preliminary results suggested that deletion of the SAC1 gene eliminated nigericin stimulated ATP efflux. However, further experimentation revealed that this result was caused by a pronounced extracellular ATPase activity for sac1delta cells at alkaline pH, conditions required to measure extracellular ATP in wild type cells. At acid pH, sac1delta cells exhibit glucose-dependent, nigericin-stimulated ATP efflux. sac1delta cells express less acid phosphatase activity in the periplasm than do wild type cells, thus increasing the stability of extracellular ATP. At alkaline pH, however, sac1delta cells tend to lose structural integrity and release lactate dehydrogenase as well as an unidentified ATPase activity to the extracellular fluid. Therefore, Sac1p is not involved in ATP efflux from S. cerevisiae. PMID- 9223425 TI - Increased MAPK expression and activity in primary human hepatocellular carcinoma. AB - We investigated the expression and activity of mitogen-activated protein kinase (MAPK) in human hepatocellular carcinoma (HCC). MAPK expression was determined in five human tumors and five normal tissues (adjacent non-neoplastic liver) by Western blotting using specific antisera raised against four MAPK pathway intermediates: Erk-1, Erk-2 (extracellular-signal regulated kinases), Mek-1 and Mek-2 (mitogen activated protein kinase kinases). There was a significant increase in Erk-1, Erk-2, Mek-1 and Mek-2 expression in particulate and cytosolic fractions prepared from tumor specimens as compared with the adjacent normal control tissues. The functional activity of both membrane and cytosolic Erk-2, determined by phosphorylation of myelin basic protein (MBP), was significantly increased in tumor specimens as compared to normal (membrane: 321%+/-50%, p<0.05; and cytosol: 597%+/-233%, p<0.05 percent of normal tissue). These data demonstrate for the first time a significant increase in MAPK expression and functional activity in human HCC. Because of the important role that the MAPK pathway plays in cellular growth and differentiation, overexpression of MAPK may be of critical importance to the formation and maintenance of human hepatocellular carcinoma. PMID- 9223426 TI - The amino-terminal region of amyloid precursor protein is responsible for neurite outgrowth in rat neocortical explant culture. AB - We have previously shown that secreted forms of beta-amyloid precursor protein (APP(s)s) promote neurite outgrowth in embryonic rat neocortical explant culture. To determine the region of APP(s) responsible for its biological activity, we produced both amino- and carboxyl-terminal regions of APP(s) using a yeast expression system. The purified fragment corresponding to the amino-terminal region (NAPP) enhanced neurite outgrowth of neocortical explants, but the carboxyl-terminal region fragment did not. The neurite-promoting activity of full length APP(s) and NAPP was blocked by the antibody, 22C11, specific for the amino terminal region, and the 16-mer peptide of epitope for 22C11 also enhanced neurite outgrowth. However, the 17-mer peptide which contains RERMS sequence did not enhance the neurite outgrowth, but promoted the survival of neocortical neurons in dissociated culture. These findings suggested that the amino-terminal region is responsible for the neurite-promoting activity of APP(s)s. PMID- 9223427 TI - Identification of heparin-binding sites in midkine and their role in neurite promotion. AB - Midkine (MK) is a heparin-binding growth factor, which promotes neurite outgrowth in embryonic neurons and enhances their survival. The three dimensional structure of MK clarified by NMR spectroscopy indicates that several basic amino acids are exposed on the surface of the C-terminal half domain, which retains heparin binding and neurite-promoting activity. We performed site-directed mutagenesis of these amino acids, and found that mutation of arginine78 reduced the heparin binding activity. Mutation of either lysine83 or lysine84 scarcely affected heparin-binding activity, while the double mutant involving both lysine residues showed reduction in the activity. Neurite-promoting activity of mutant MKs always correlated with their heparin-binding activity, illustrating the close relationship of the two activities. Thus, the present result verifies the occurrence of two distinct heparin-binding sites involved in neurite-promoting activity of MK molecule. PMID- 9223428 TI - Increase in Ref-1 mRNA and protein by thyrotropin in rat thyroid FRTL-5 cells. AB - Thyrotropin (TSH) induces the expression of fos and jun family genes in thyroid cells. The DNA-binding activity of these gene products (AP-1) has been shown to be enhanced by ubiquitous nuclear redox factor-1 (Ref-1). We thus examined whether TSH regulates Ref-1 gene expression in rat thyroid FRTL-5 cells. Northern blot analysis revealed that the abundance of Ref-1 mRNA significantly increased within 3 hours after TSH followed by a sustained increase until 12 hours. The increase was also induced by treatment with forskolin but not in the presence of cycloheximide, indicating that the TSH effect on Ref-1 mRNA is mediated by intracellular cAMP and requires de novo protein synthesis. Consistent with the elevation of the mRNA level, Western blot analysis showed an increase in Ref-1 protein 3 hours after TSH. The level continued to increase until 12 hours. These results suggested that increased Ref-1 by TSH might regulate the binding activity of AP-1 in thyroid cells. Considering that Ref-1 also has a DNA repair function, Ref-1 may play dual roles in gene regulation and DNA repair processes in thyroid cells. PMID- 9223429 TI - A novel fungal gene encoding chitin synthase with a myosin motor-like domain. AB - A csmA gene that encodes chitin synthase with a myosin motor-like domain was isolated from the filamentous fungus Aspergillus nidulans. Initially, we obtained the csmA as a homolog of the Aspergillus fumigatus chsE-partial fragment. A large open reading frame encoding a polypeptide of 1,852 a.a. was identified by determining the cDNA sequences. The chitin synthase conserved region was situated at the C-terminus and classified into class V as reported previously. On the other hand, the N-terminal region showed significant similarity to myosin motors and could not be classified into any types of myosins identified so far. Thus, it is suggested that this is the first report of unconventional myosin fused to a metabolic enzyme. The finding of this new type of chitin synthase gene suggests that localization of chitin synthesis may be guided by association with cytoskeletal structures. PMID- 9223431 TI - A splicing variant of Steroid Receptor Coactivator-1 (SRC-1E): the major isoform of SRC-1 to mediate thyroid hormone action. AB - Steroid Receptor Coactivator-1 (SRC-1) interacts with nuclear receptors only when they are bound to the ligands and enhance the transactivation. We identified splicing variants encoding three isoforms, SRC-1, SRC-1(-Q), and SRC-1E, generated by alternative usage of an exon(s) and splicing acceptor sites. RT-PCR analysis showed that SRC-1E was more abundantly expressed than SRC-1 or SRC-1(-Q) at the mRNA level in all the cell lines tested. SRC-1E lacks 56 amino acids of SRC-1 and has unique 14 amino acids at the carboxyl terminus, while SRC-1(-Q) differs from SRC-1 by deletion of only one glutamine residue. Since the C terminal domain of SRC-1 has been shown to be involved in the interaction with nuclear receptors, the enhancement of transactivation by these three isoforms was tested. SRC-1E enhanced thyroid hormone dependent transactivation of reporter gene expression more profoundly than SRC-1 or SRC-1(-Q). Taken together, it was suggested that SRC-1E is the major isoform of SRC-1 to mediate thyroid hormone action. PMID- 9223430 TI - Activation of the p21(Waf1/Cip1) promoter by the ets oncogene family transcription factor E1AF. AB - p21(Waf1/Cip1) is one of the key regulatory proteins in cell cycle, terminal differentiation, and apoptosis. Its promoter was shown to be transactivated by the wild-type p53 protein as well as in a p53-independent manner. In this report, we demonstrate that E1AF, an ets-related transcription factor, activates the human p21(Waf1/Cip1) promoter by interacting with the ets-binding sites located close to the two previously identified p53-responsive elements. Northern blot analysis revealed that p21(Waf1/Cip1) and E1AF were correlatively upregulated in response to cisplatin treatment in SiHa cells. Transient expression assays demonstrated that E1AF can activate the p21(Waf1/Cip1) promoter-driven luciferase reporter gene in SiHa cells. The p21(Waf1/Cip1) promoter activity was also increased in p53-null Saos2 osteosarcoma cells, but was markedly reduced when the ets-binding sites were deleted. These results indicate that E1AF positively regulates transcription from the p21(Waf1/Cip1) promoter in response to genotoxic stresses. PMID- 9223432 TI - How important is the N-3 sugar moiety in the tight-binding interaction of coformycin with adenosine deaminase? AB - Preliminary findings on the possible important role of the N-3 sugar moiety of coformycin in its tight-binding interaction with adenosine deaminase (ADA) are reported. The compound 3-beta-D-Ribofuranosyl-5,6,7,8-tetrahydro-4H-imidazo[4,5 d][1,3]diaze pin-5-one-8-ol (1), its 3-benzyl analogue (6), and the aglycon (7) served as probes. The first two were both found to be competitive inhibitors of ADA with Ki's in the range of 10(-5) M, while the last one was inactive. PMID- 9223433 TI - Partial purification and identification of hormone-sensitive lipase from chicken adipose tissue. AB - HSL from chicken adipose tissue exhibits remarkable activation upon phosphorylation with cAMP-dependent protein kinase (cAMP-PK) compared to HSL from rat and human adipose tissue. In order to characterize the chicken HSL enzyme, it was purified 3500 fold from a chicken adipose tissue homogenate using pH 5.2 precipitation and anion-exchange chromatography. The purified chicken HSL was identified as an 86 kDa protein using Western blot analysis. The HSL diacylglycerol lipase activity was inhibited by 98% upon incubation with anti-rat HSL antiserum, and the specific activity of chicken HSL was estimated to be approximately the same as for the rat enzyme. Furthermore, the 86 kDa polypeptide was phosphorylated by cAMP-PK to about the same stoichiometry as for the recombinant rat enzyme. Hence, our results demonstrate that HSL from chicken adipose tissue is comparable in size and specific activity to HSL from mammalian species, and not a smaller 42 kDa polypeptide with 1000-fold lower specific activity as previously reported (Berglund, L., Khoo, J. C., Jensen, D., and Steinberg, D., 1980 J. Biol. Chem. 255, 5420-5428). PMID- 9223434 TI - The C-terminal domain of tissue inhibitor of metalloproteinases-2 is required for cell binding but not for antimetalloproteinase activity. AB - We have generated a C-terminally-truncated form of recombinant tissue inhibitor of metalloproteinases-2 (designated rTIMP-2 delta) in which the region of the inhibitor extending from residue 128 to 194 and including 3 of the 6 disulfide bonds is deleted. rTIMP-2 and rTIMP-2 delta had similar inhibitory activities toward interstitial collagenase and inhibited the activation of the precursor form of matrix metalloproteinase-2 (proMMP-2). rTIMP-2 also bound with high affinity (Kd 0.99 nM) to HT1080 human fibrosarcoma cells treated with 12-O tetradecanoyl-phorbol-13-acetate. However deletion of the C-terminal domain of TIMP-2 significantly lowered the cell surface binding affinity, with competition experiments indicating a 2 order of magnitude difference between rTIMP-2 and rTIMP-2 delta in the concentrations needed to displace 125I-labeled rTIMP-2 binding. These data indicate that the C-terminal domain of TIMP-2 is not required for the antimetalloproteinase activity but plays a major role in the high affinity cell surface binding of the inhibitor. PMID- 9223435 TI - Cloning of a human heptahelical receptor closely related to the P2Y5 receptor. AB - The 6H1 receptor cloned from activated chicken T cells was initially considered an orphan G-coupled receptor, but was later included in the P2Y family of receptors for purine and pyrimidine nucleotides on the basis of a significant amino acid identity and was renamed P2Y5. Analysis of the expressed sequence tag database revealed the presence of a related sequence exhibiting 63% amino acid identity with this receptor. Starting from this partial sequence, we have isolated a complete clone and identified a 1113 base pair open reading frame encoding a new G-coupled receptor that we have called P2Y5-like. This sequence exhibits 61% identity with the chicken P2Y5 sequence and 30-33% with other P2Y subtypes. A construct encoding this P2Y5-like receptor was transfected into COS 7, 1321N1, and CHO-K1 cells, and expression was documented by Northern blotting. None of the 40 nucleotides and nucleosides tested was able to elicit a response in any of four functional assays: inositol phosphate formation, stimulation or inhibition of cAMP formation, and extracellular acidification measured with a microphysiometer. These results suggest either that the natural ligand of the P2Y5-like receptor is an uncommon nucleotide or alternatively that despite its structural similarity to the P2Y family it is not a nucleotide receptor. PMID- 9223436 TI - Localization of an exchange inhibitory peptide (XIP) binding site on the cardiac sodium-calcium exchanger. AB - The exchange inhibitory peptide (XIP; RRLLFYKYVYKRYRAGKQRG) is a potent inhibitor of cardiac Na-Ca exchange activity. This study attempted to identify the XIP binding site on the Na-Ca exchange protein. Bovine cardiac sarcolemmal vesicles were proteolyzed and fractionated by XIP-affinity column chromatography. A 24 kDa fragment was purified and subjected to amino acid sequence analysis. A negatively charged region of intracellular loop f of the Na-Ca exchange protein (IDDDIFEEDEN; aa 445-455) was identified. The affinity and specificity of XIP interaction with peptides IDDDIFEEDEN and GEDDDDDECGEE (another negatively charged region of the Na-Ca exchange protein) were examined. XIP cross-linked to peptide IDDDIFEEDEN but not GEDDDDDECGEE in a pH-dependent manner. Fluorescence titration binding studies indicated that binding of IDDDIFEEDEN with XIP was saturable (Kd=5 microM) while binding with GEDDDDDECGEE was not specific. These data suggest that amino acids 445-455 on Na-Ca exchange loop f are involved in XIP binding. PMID- 9223438 TI - A cationic lipid for rapid and efficient delivery of plasmid DNA into mammalian cells. AB - Cationic lipids are widely used for gene transfer into cultured eukaryotic cells. However, lipids with potent transfection activity are often associated with high levels of cytotoxicity, and also require serum-free conditions for optimal performance. These characteristics in many cases result in unsatisfactory transfection efficiency. In this report, we describe a new cationic amphiphile, N t-butyl-N'-tetradecyl-3-tetradecylaminopropionamidine (Amidine). Amidine requires only 1-2 hour incubation intervals to produce maximal transfection efficiency, and can transfect cells in the presence of serum. Such characteristics significantly minimize cytotoxicity, and also provide time flexibility for researchers. We routinely obtain over 80% transfection efficiency as evidenced by use of an enhanced green fluorescence protein (EGFP) as the reporter. These studies demonstrate the utility of Amidine for rapid and efficient transfection of mammalian cells. PMID- 9223437 TI - Expression, nuclear localization and interactions of human MCM/P1 proteins. AB - We report here the comparative analysis of human Mcm/P1 proteins (HsMcm2, -3, -5 and -7), including a characterization of their mutual interactions, cell cycle dependent expression and nuclear localization during the cell cycle and the quiescent state. The mRNA levels of these genes, which undergo cell cycle dependent oscillations with a peak at G1/S phase, may be regulated by E2F motifs, two of which were detected in the 5' upstream region of the HsMCM5 gene. In contrast, the protein levels of these Mcm proteins were found to remain rather constant during the HeLa cell cycle. However, their levels gradually increased in a variable manner as KD cells progressed from GO into the G1/S phase. In the GO stage, the amounts of HsMcm2 and -5 proteins were much lower than those of HsMcm7 and -3 proteins, suggesting that they are not present in stoichiometric amounts, and that only a proportion of these molecules actively participate in cell cycle regulation as part of Mcm/P1 complexes. PMID- 9223439 TI - Evidence for a mammalian Nim1-like kinase pathway acting at the G0-1/S transition. AB - In fission yeast the Nim1 kinase phosphorylates and inactivates the cdc2 inhibitory Weel tyrosine kinase. In addition, Nim1 is necessary for an efficient cellular response to nutritional starvation leading to cell cycle arrest in G1. Given the remarkable evolutionary conservation of the cell cycle regulatory mechanism we have investigated the effect of Nim1 expression on the control of the mammalian cell cycle using a plasmid microinjection approach. In synchronised IMR90 human fibroblasts, expression of Nim1 strongly inhibited entry into S phase. This effect was dependent on the catalytic activity of Nim1 and did not require its regulatory domain. Furthermore we show that co-expression of human Wee1 kinase reverted the inhibitory effect, indicating that Nim1 was acting in a Wee1-dependent manner. These results provide evidence for the existence of a Nim1 like kinase pathway acting at the G0-1/S transition in human cells. PMID- 9223440 TI - Expression and regulation of transforming growth factor beta1 mRNA and protein in rat fetal testis in vitro. AB - The expression and secretion of Transforming Growth Factor beta1 (TGFbeta1) by cultured testes of day 20.5 rat fetuses were investigated. The testes were found to express two TGFbeta1 mRNA transcripts of 2.5 and 1.8 kb. By using mink lung epithelial cell bioassay based on the measurement of the inhibition of tritiated thymidine incorporation in response to TGFbeta1 immunoreactive material, the fetal testes were shown to secrete TGFbeta1 protein in organ culture. This secretion was positively regulated by dibutyryl cyclic AMP or by LH and FSH together, but not by LH alone and very slightly by FSH alone, which suggests interactions between Leydig and Sertoli cells for the control of TGFbeta1 production. These regulations probably take place at a posttranscriptional step since no concomitant increase of TGFbeta1 mRNA levels was observed. Such a positive regulation of TGFbeta1 secretion by gonadotropins could be a characteristic of the rat fetal testis. PMID- 9223441 TI - Agonistic effect of tamoxifen is dependent on cell type, ERE-promoter context, and estrogen receptor subtype: functional difference between estrogen receptors alpha and beta. AB - To investigate the functional differences between estrogen receptor (ER) alpha and beta subtypes, we studied the expression and the transcription stimulating activities of these receptors. RT-PCR has demonstrated that ER alpha is expressed at a high level in MCF-7 cells derived from human breast cancer. Both ER alpha and ER beta were expressed at a lower level in HOS-TE85 and Saos2 cells derived from human osteosarcoma. Chloramphenicol acetyltransferase reporter assay detected the transcriptional activation by the endogenous receptor only in MCF-7 cells. Agonistic effect of tamoxifen was observed as strong as that of 17beta estradiol on ERE activation in MCF-7 cells at the concentration of 10(-7) M when ERE-containing reporter is constructed with beta-globin promoter. The effect of tamoxifen was not apparent when the reporter was constructed with thymidine kinase promoter, suggesting that the differential gene activation between tamoxifen and estrogen may take place depending upon ERE-promoter context. Agonistic activity of tamoxifen was also detected in COS-7 and Saos-2 cells, but not in HEC-1 cells derived from human endometrial carcinoma via exogenously expressed ER. Interestingly, this effect was ER alpha specific. Thus, we demonstrate that agonistic effect of tamoxifen depends on the cell type, ERE promoter context, and ER subtype. These parameters would explain at least a part of the tissue specific effects of antiestrogens in vivo. PMID- 9223442 TI - Comparison of a left-handed Z-DNA molecular structure determined by X-rays with that simulated by a molecular dynamics. AB - The 1.0 A resolution X-ray crystal structures of the left-handed Z-DNA(Z-I and Z II conformations) were compared with that of the simulated molecular dynamics(MD) structure both in vacuo and in solution. Whilst the X-ray structure showed a tendency for the d(CG)3 molecule to take on a Z-II conformation in high salt solution or in strongly ionized conditions, the MD simulation with Na ion for 30 ps revealed that the left-handed d(CG)3 structure with the Z-I conformation was transformed into the Z-II conformation in the torsion angles of the C3, G4 and C5 phosphate groups, and furthermore, when K+ ion was used as the counterion instead of Na+ ion, the torsion angles in almost the entire d(CG)3 molecule were preserved. On the other hand, the MD calculation resulted in some very important changes on the sugar puckerings; the simulation with Na+ ion indicated that all the sugar puckerings of cytosine residues were changed from C2'-endo to C3'-endo, while those for guanosine residues tended to keep unchanged (C3'-endo) except for a terminal residue (G6). PMID- 9223443 TI - Down-regulation of the defective transcripts of the Werner's syndrome gene in the cells of patients. AB - Werner's syndrome (WS), an adult progeria, is a recessive genetic disorder caused by the mutations in the DNA helicase gene (WRN). In this study, a comparative northern blot analysis was made for poly(A)+ RNAs extracted from fibroblasts and B-lymphoblastoid cells of WS patients, relatives of patients, and normal individuals. The levels of mutant WRN mRNA from patient cells were significantly lower than those of intact mRNA from the cells of normal individuals by an average of 70%. Furthermore, an extremely low level of WRN mRNA(s), presumably a mixture of mutant and intact mRNAs, was observed for the patient's family members who carry one mutated allele. These results strongly suggest that a relatively low level of helicase mRNA is sufficient to prevent the onset of Werner's syndrome. PMID- 9223444 TI - Caveolin-1 detergent solubility and association with endothelial nitric oxide synthase is modulated by tyrosine phosphorylation. AB - Caveolin-1 and endothelial nitric oxide synthase (eNOS) are associated within endothelial caveolae. We have shown previously that eNOS is translocated to the detergent-insoluble, cytoskeletal fraction of bovine aortic endothelial cells (BAEC) in response to bradykinin (BK)-stimulation or tyrosine phosphatase inhibition. In the present study, we have examined whether caveolin-1 is similarly translocated in response to these or other stimuli. Exposure of BAEC to the eNOS-activating agonists, BK, histamine, or ATP produces transient increases in the amounts of detergent-insoluble caveolin-1. Increases in insolubility are blocked by tyrosine kinase inhibitors and are potently mimicked by tyrosine phosphatase inhibitors. Increased insolubility is accompanied by an increased association of caveolin-1 with eNOS and inhibition of eNOS catalytic activity. Agonist-activation of eNOS in endothelial cells thus appears to involve tyrosine phosphorylation-dependent changes in the interaction of eNOS with caveolin-1. Increased interaction of eNOS with caveolin-1 may deactivate the enzyme subsequent to its activation by Ca2+/calmodulin. PMID- 9223445 TI - Insulin secretagogues with an imidazoline structure inhibit arginine-induced secretion from isolated glucagon secretion from isolated rat islets of Langerhans. AB - It is well documented that imidazoline compounds such as efaroxan and phentolamine act as potent insulin secretagogues both in vivo and in vitro, an effect which is mediated principally by blockade of ATP-sensitive potassium channels in the pancreatic B-cell. However, little is known about the effects of these drugs on the secretion of other pancreatic hormones and, in the present work, we have investigated the effects of selective imidazoline compounds on glucagon release from isolated rat islets of Langerhans. None of several imidazoline compounds tested (efaroxan, phentolamine, idazoxan, antazoline) affected glucagon secretion from islets incubated with 4 mM glucose. However, when the rate of glucagon release was stimulated by L-arginine (20 mM) efaroxan caused a rapid, sustained and dose-dependent inhibition of the secretory response (EC50 approximately 30 microM). This effect was seen under both static incubation and islet perifusion conditions. Antazoline and phentolamine also inhibited arginine-induced glucagon secretion, whereas idazoxan (an imidazoline which does not affect insulin secretion) failed to alter glucagon release. The inhibitory effects of imidazolines on glucagon release were not secondary to changes in insulin secretion. Taken together, the results indicate that pancreatic A-cells express functional imidazoline receptors which can regulate the secretory activity of the cells. PMID- 9223446 TI - Cultivation, serial transfer, and differentiation of epidermal keratinocytes in serum-free medium. AB - We describe serum-free culture conditions for human epidermal keratinocytes using lethally treated 3T3 cells as feeder layers and normal Ca++ concentrations (1.2 mM), in a DMEM/F12-Ham nutrient mixture supplemented with several additives, and 10 mg/ml bovine serum albumin instead of animal serum. Keratinocytes were serially grown to 15-18 cell generations (4 subcultivations) and formed a stratified squamous epithelium that could be detached as a graftable epithelial sheet. EGF and TGF alpha significantly increased keratinocyte proliferation under these conditions; EGF reduced the expression of keratin K1, which is specific for stratified and terminally differentiated epidermal keratinocytes. In contrast with previous reports, the serum-free medium we describe here supports serial growth and normal differentiation of human epidermal keratinocytes, and the formation of graftable stratified epithelia; it also supports the assay of a variety of cytokines or compounds that modulate epidermal keratinocyte proliferation and differentiation. PMID- 9223447 TI - Qualitative and quantitative catalog of tyrosinase alternative transcripts in normal murine skin melanocytes as a basis for detecting melanoma-specific changes. AB - The decline in cell differentiation commonly associated with malignant progression may be due in part to an increase in alternative splicing of the pre mRNAs of tissue-specific genes. As a necessary basis for investigating this possibility in a murine model of cutaneous melanoma, the complete qualitative and quantitative inventory of alternative transcripts was sought for the tyrosinase gene in normal mouse skin melanocytes, as this gene plays a key role in melanization. Of 111 alternative mRNAs predicted from known splice sites in the gene, 19 isoforms were detected, and their abundances determined, through a systematized protocol involving splice junction-specific probes, exon-specific restriction enzymes, and quantitative RT-PCR with an RNA internal standard. No unpredicted tyrosinase transcripts were discovered. Two of the transcripts, each involving an intra-exonic deletion and present at relatively low abundance in normal skin, were subsequently found to be consistently upregulated in melanomas. PMID- 9223448 TI - cDNA cloning of a novel amphiphysin isoform and tissue-specific expression of its multiple splice variants. AB - Amphiphysin, an SH3-domain containing protein concentrated in nerve terminals, is believed to be involved in the synaptic vesicle recycling. We have cloned cDNAs of a novel isoform of amphiphysin (amphiphysin II) by exploiting sequence information for homologous ESTs deposited in databases. At least 9 different subtypes of the isoform with 50-60% amino acid identity to the human amphiphysin were identified by a conventional library screening and PCR amplification of cDNA libraries. Each subtype probably represents a splice variant of a single gene transcript. Analysis of mRNA expression in various tissues by RT-PCR showed that the isoform is ubiquitously distributed. The expression spectrum of the isoform subtypes, however, is significantly different in several tissues examined, suggesting that they are involved in the regulation of endocytic processes that are unique to each cell type. PMID- 9223449 TI - Evidence that synaptobrevin is involved in fusion between synaptic vesicles and synaptic plasma membrane vesicles. AB - We have developed a model system, consisting of rat brain synaptic vesicles and rat brain synaptic plasma membrane vesicles, to study the fusion process associated with the exocytotic release of neurotransmitters. Our results show a significant increase in the extent of fusion when the reaction takes place in cytosol compared to that obtained when fusion is carried out in buffer. This effect is mediated by cytosolic proteins, although N-ethylmaleimide-sensitive factor does not play a role in fusion. We also registered an almost complete inhibition of fusion when synaptic vesicles were pre-incubated with botulinum toxin B, indicating that synaptobrevin plays an important role in the coalescence of membrane lipids of the interacting membranes. PMID- 9223450 TI - Increased expression of caveolin-1 in heterozygous Niemann-Pick type II human fibroblasts. AB - Human Niemann-Pick type II fibroblasts, which encompass the panethnic type C (NPC) and Nova Scotia Acadian type D (NPD) variants, exhibit altered expression of caveolin-1 protein when examined by immunoblotting using an anti-caveolin-1 monoclonal antibody. Unexpectedly, caveolin-1 in heterozygous fibroblasts was significantly elevated as much as 10-fold compared to caveolin-1 in normal and homozygous affected fibroblasts. Homozygous NPC fibroblasts expressed caveolin-1 levels similar to those in normal fibroblasts, while the expression of caveolin-1 in homozygous NPD fibroblasts was slightly elevated. Northern analysis indicates that normal fibroblasts and NPC heterozygous fibroblasts have similar amounts of caveolin-1 mRNA, while NPC homozygous fibroblasts have significantly less caveolin-1 mRNA. In contrast, heterozygous and homozygous NPD fibroblasts exhibit increased levels of caveolin-1 mRNA. These novel findings suggest that caveolin-1 containing subcellular structures are involved in the pathophysiology of Niemann Pick type II disease. Furthermore, altered caveolin-1 protein expression may serve as a useful marker for the diagnosis of carriers of NPC or NPD. PMID- 9223451 TI - Interleukin-1beta-converting enzyme and CPP32 are involved in ultraviolet B induced apoptosis of SV40-transformed human keratinocytes. AB - Interleukin-1beta-converting enzyme (ICE) and CPP32 are cysteine proteinases, that are involved in apoptotic process in various cell systems. We investigated the effects of ICE on ultraviolet B (UVB) induced-apoptosis in SV40-transformed human keratinocytes (SVHK cells). The ICE inhibitor (Z-Val-Ala-Asp-CH2F) and CPP32 inhibitor (Z-Asp-Glu-Val-Asp-CH2F) blocked the apoptotic cell death caused by UVB irradiation. The addition of both ICE and CPP32 inhibitors to the incubation medium resulted in neither an additive nor a synergistic suppression of UVB-induced apoptosis. Reverse transcription and polymerase chain reaction (RT PCR) analysis indicated that SVHK cells expressed ICE-alpha, and beta mRNAs. UVB irradiation increased the mRNA of both isoforms and Western blot analysis confirmed that UVB increased an active form of ICE protein, p20, that is generated by autoproteolytic cleavage of inactive 45 kDa proenzyme derived from both mRNAs. Transfection of ICE expression vector into SVHK cells resulted in apoptosis in a dose dependent manner and UVB-irradiation further augmented the ICE expression vector-induced apoptosis. These results indicate that ICE plays an important role in UVB-induced apoptosis of SVHK cells. PMID- 9223452 TI - Extracellular signal-regulated kinase and c-Jun NH2-terminal kinase activities are continuously and differentially increased in aorta of hypertensive rats. AB - We first examined the activities of extracellular signal-regulated kinases (ERKs) and c-Jun NH2-terminal kinases (JNKs) in the aorta of hypertensive rats. In Dahl salt-sensitive (DS) rats, chronic hypertension caused by a high-salt diet was followed by sustained activation of aortic p42ERK and p44ERK. p46JNK and p55JNK activities were also increased in hypertensive DS rats, but returned to control levels earlier than ERKs, suggesting that ERKs and JNKs may be independently activated in hypertensive rats. In stroke-prone spontaneously hypertensive rats (SHRSP) which spontaneously develop hypertension under a low salt-diet, aortic p42ERK and p44ERK activities were progressively increased with the development of hypertension, compared with control normotensive rats. p46JNK and p55JNK activities in SHRSP were increased, with a different time course from ERKs. Thus, we first demonstrated that ERKs and JNKs activities are chronically and differentially increased in the aorta of hypertensive rats, suggesting the involvement of these kinases in hypertensive vascular diseases. PMID- 9223453 TI - Factors altering ribozyme-mediated cleavage of tumor necrosis factor-alpha mRNA in vitro. AB - Hammerhead ribozymes are capable of cleaving RNA in a sequence specific manner in vitro. However, the complex environment of the cell differs dramatically from the conditions in vitro. Therefore, we explored cleavage of full-length target RNA with two ribozymes targeted against the murine tumor necrosis factor-alpha (TNF alpha) mRNA. These ribozymes cleaved TNF-alpha mRNA within a pool of total cellular RNA in vitro, but less efficiently than previously reported for similar ribozymes. Although there may be several factors that could affect ribozyme activity, two of these factors were tested. The first factor was whether non target polynucleotides inhibited ribozyme-mediated cleavage. Total cellular RNA and to a lesser degree DNA inhibited ribozyme activity. This inhibition was a combination of competitive and non-competitive inhibition. Non-target RNA with minimal complementarity to the ribozyme or target showed no effect on cleavage rates. The second factor was whether denaturing conditions improved ribozyme cleavage efficiency. Hammerhead ribozymes with 24 complementary bases had increased cleavage efficiency in formamide. Thus, the ribozymes may have had too long of an antisense flanking sequence which hybridized with the target RNA and resulted in a high melting temperature. These studies demonstrate that ribozyme cleavage was influenced by the amount of non-target polynucleotide and the strength of the ribozyme-substrate interaction. PMID- 9223454 TI - STRL22 is a receptor for the CC chemokine MIP-3alpha. AB - STRL22 is a human seven transmembrane domain orphan receptor related to known chemokine receptors and expressed in peripheral blood lymphocytes, tumor infiltrating lymphocytes and lymphoid tissues. MIP-3alpha/LARC/Exodus is a CC chemokine that is chemotactic for lymphocytes and that is expressed in activated cells, including monocytes, T cells, endothelial cells, and fibroblasts, and in liver, lung, and some lymphoid tissues. We report here that STRL22-transfected human embryonic kidney 293 cells demonstrated specific binding for MIP-3alpha and that MIP-3alpha, but no other chemokines, produced a calcium flux in the STRL22 transfected cells. We show that MIP-3alpha, unlike other chemokines, produced a calcium flux in freshly-isolated peripheral blood lymphocytes and we show that MIP-3alpha also produced a signal in tumor infiltrating lymphocytes that express STRL22. Since STRL22 is the sixth functional CC chemokine receptor identified, it should be re-named CCR6. PMID- 9223455 TI - A close association of GM3 with c-Src and Rho in GM3-enriched microdomains at the B16 melanoma cell surface membrane: a preliminary note. AB - B16 melanoma is characterized by high content of GM3 ganglioside, which has been recognized as a melanoma-associated antigen defined by specific monoclonal antibodies. We report now that GM3 is present predominantly (>90%) in the 1% Triton X-100-insoluble, low-density microvesicular fraction ("detergent-insoluble glycosphingolipid-enriched microdomain"; DIGEM) separated on sucrose density gradient centrifugation. Associated with DIGEM, many signal transducer molecules such as c-Src, FAK, and the low-molecular-weight G-proteins Rho A and H-Ras were also found. Rho A and FAK were found in part, and PLC-beta2 and G alphas were found exclusively, in the high-density fraction. Immunoprecipitation of GM3 present in DIGEM by anti-GM3 antibody DH2, followed by Western blotting, revealed co-precipitation of Rho A and c-Src with GM3. These findings suggest (i) a specific organization of GM3 in close association with Rho A and c-Src within DIGEM at the melanoma cell surface; and (ii) such organizational units may be directly involved in signal transduction, in which glycosphingolipids receive signals which are subsequently transduced by associated transducer molecules. PMID- 9223456 TI - Extracellular ATP triggers cyclic AMP-dependent differentiation of HL-60 cells. AB - Extracellular ATP and ATPgammaS (1-1000 microM) stimulated cyclic AMP (cAMP) production in undifferentiated HL-60 cells. The potency order for adenine nucleotides and adenosine was ATPgammaS > ATP >> ADP > or = AMP = Adenosine. Indomethacin (50 microM) had no effect on ATP-induced cAMP production. ATP and ATPgammaS also suppressed cell growth and induced differentiation as revealed by fMLP-stimulated beta-glucuronidase release 48 h after exposure. The potency order for the induction of fMLP-stimulated beta-glucuronidase release by adenine nucleotides and adenosine was ATPgammaS > or = ATP > ADP > AMP = Adenosine approximately 0. The protein kinase A inhibitor Rp-8-Br-cAMPS (10-200 microM) suppressed ATP-induced differentiation but had no effect on ATP-dependent growth suppression. UTP which, like ATP, activates P2U receptors on HL-60 cells, had no effect on cAMP production, cell growth, or differentiation. The data suggest the existence of a novel receptor for ATP on undifferentiated HL-60 cells that is coupled to the activation of adenylate cyclase and cAMP-dependent differentiation. PMID- 9223457 TI - Mechanism of human immunodeficiency virus type 1 localization in CD4-negative thymocytes: differentiation from a CD4-positive precursor allows productive infection. AB - Human immunodeficiency virus (HIV) infection of the thymus could have profound effects on development of the immune response, particularly in children. We and others have established that in addition to infecting and depleting CD4-bearing thymocytes, functional HIV proviruses are found in thymocytes lacking surface CD4 expression. Using in vitro thymocyte cultures, we show that neither HIV-mediated down regulation of CD4 nor CD4-independent infection contributes to the localization of HIV in cells lacking the primary virus receptor. Rather, infection of a CD4-positive precursor cell (CD4 positive/CD8 positive) with subsequent differentiation into a mature CD4-negative phenotype results in productively infected CD4-negative cells. This novel mechanism may contribute to pathogenesis by distributing viral sequences into functional subsets of T cells typically refractory to HIV infection and could account for the presence of viral DNA in CD8-positive lymphocytes recently observed in patients. PMID- 9223459 TI - Parvovirus initiation factor PIF: a novel human DNA-binding factor which coordinately recognizes two ACGT motifs. AB - A novel human site-specific DNA-binding factor has been partially purified from extracts of HeLa S3 cells. This factor, designated PIF, for parvovirus initiation factor, binds to the minimal origin of DNA replication at the 3' end of the minute virus of mice (MVM) genome and functions as an essential cofactor in the replication initiation process. Here we show that PIF is required for the viral replicator protein NS1 to nick and become covalently attached to a specific site in the origin sequence in a reaction which requires ATP hydrolysis. DNase I and copper ortho-phenanthroline degradation of the PIF-DNA complexes showed that PIF protects a stretch of some 20 nucleotides, covering the entire region in the minimal left-end origin not already known to be occupied by NS1. Methylation and carboxy-ethylation interference analysis identified two ACGT motifs, spaced by five nucleotides, as the sequences responsible for this binding. A series of mutant oligonucleotides was then used as competitive inhibitors in gel mobility shift assays to confirm that PIF recognizes both of these ACGT sequences and to demonstrate that the two motifs comprise a single binding site rather than two separate sites. Competitive inhibition of the origin nicking assay, using the same group of oligonucleotides, confirmed that the same cellular factor is responsible for both mobility shift and nicking activities. UV cross-linking and relative mobility assays suggest that PIF binds DNA as a heterodimer or higher order multimer with subunits in the 80- to 100-kDa range. PMID- 9223458 TI - Determinants of the human immunodeficiency virus type 1 p15NC-RNA interaction that affect enhanced cleavage by the viral protease. AB - During human immunodeficiency virus type 1 (HIV-1) virion assembly, cleavage of the Gag precursor by the viral protease results in the transient appearance of a nucleocapsid-p1-p6 intermediate product designated p15NC. Utilizing the p15NC precursor protein produced with an in vitro transcription-translation system or purified after expression in Escherichia coli, we have demonstrated that RNA is required for efficient cleavage of HIV p15NC. Gel mobility shift and nitrocellulose filter binding experiments indicate that purified p15NC protein specifically binds its corresponding mRNA with an estimated Kd of 1.5 nM. Binding was not affected by the presence or absence of zinc or EDTA. Moreover, mutagenesis of the cysteine residues within either of the two Cys-His arrays had no effect on RNA binding or on RNA-dependent cleavage by the viral protease. In contrast, decreased binding of RNA and diminished susceptibility to cleavage in vitro were observed with p15NC-containing mutations in one or more residues within the triplet of basic amino acids present in the region between the two zinc fingers. In addition, we found that 21- to 24-base DNA and RNA oligonucleotides of a particular sequence and secondary structure could substitute for p15 RNA in the enhancement of p15NC cleavage. Virus particles carrying a mutation in the triplet of NC basic residues (P3BE) show delayed cleavage of p15NC and a defect in core formation despite the eventual appearance of fully processed virion protein. These results define determinants of the p15NC RNA interaction that lead to enhanced protease-mediated cleavage and demonstrate the importance of the triplet of basic residues in formation of the virus core. PMID- 9223460 TI - Blocking of feline immunodeficiency virus infection by a monoclonal antibody to CD9 is via inhibition of virus release rather than interference with receptor binding. AB - A monoclonal antibody, MAb vpg15, inhibits feline immunodeficiency virus (FIV) infection in tissue culture. The antibody is directed to a determinant of the feline cell surface marker, CD9, implying that CD9 may serve as a viral receptor or coreceptor in this system. In cells expressing CD9, MAb vpg15 markedly delayed acute virus infection in terms of reverse transcriptase activity detected in cell culture supernatants. This effect was evident if the antibody was added before, immediately after, or 24 h after virus infection. Binding experiments showed that MAb vpg15 did not block virus binding to the cells. PCR analyses at various intervals postinfection also indicated that MAb vpg15 did not block virus uptake, reverse transcription of viral RNA, or integration into host cell DNA. Multiply spliced mRNAs were detected up to 24 h postinfection in both control and MAb vpg15-treated cells. However, viral mRNAs were markedly diminished in MAb vpg15 treated cells after this time, consistent with a failure of the FIV infection to spread in the cell culture. Treatment of chronically infected cells with MAb vpg15 also caused a sharp diminution in viral particle production, while viral mRNA levels were the same in both untreated and MAb-treated infected cells. Analyses of intracellular and extracellular levels of virus-associated antigens showed an enhanced accumulation of intracellular p24. These findings are consistent with the interpretation that MAb vpg15 acts at a posttranscriptional stage by interfering with the assembly and/or release of virus from the cell. PMID- 9223461 TI - The importance of the A-rich loop in human immunodeficiency virus type 1 reverse transcription and infectivity. AB - Nucleotide segment (+169)AAAA(+172) constitutes an A-rich loop within human immunodeficiency virus type 1 (HIV-1) (HXB2D) RNA and is able to interact with the anticodon loop (33)/USUU(36) of primer tRNA3(Lys). We have shown that the deletion of this A-rich loop resulted in diminished levels of infectivity and reduced synthesis of viral DNA in MT-2 cells and cord blood mononuclear cells. Endogenous reverse transcriptase (RT) assays revealed that the mutated viruses, termed HIV/del-A, generated fewer cDNA products than did wild-type virus, designated HIV/WT. We also employed in vitro RT assays with in vitro-synthesized viral RNA templates, recombinant HIV-1 RT(p66/51), and natural tRNA3(Lys) as primers to show that the mutated RNA templates, designated PBS/del-A, generated less minus-strand strong-stop DNA product than did the wild-type RNA template, designated PBS/WT. The initiation efficiency of reverse transcription from the mutated RNA template was significantly impaired compared with that from the wild type RNA template when a single-base extension assay from the tRNA3(Lys) primer was employed. However, RT reactions performed with DNA oligonucleotides complementary to the primer binding site (PBS) as primers did not yield differences between the mutated PBS/del-A and wild-type RNA templates. Long-term culture of HIV/del-A in MT-2 cells resulted in the replacement of two G's at nucleotide positions 167 and 168 by two A's that possessed the same relationship to the 5' end of the PBS as did the wild-type A's at positions 171 and 172. In vitro RT assays performed with recombinant enzyme with tRNA3(Lys) as the primer showed that the RNA template thus generated, termed PBS/A2, yielded levels of minus-strand strong-stop DNA product similar to those yielded by the wild-type RNA template. Coincidentally, viruses containing A's at positions 167 and 168 were able to replicate with efficiencies similar to those of the wild-type viruses. Thus, the (+169)AAAA(+172) A-rich loop plays a key role in the synthesis of viral DNA. PMID- 9223462 TI - The 85-kilodalton phosphoprotein (pp85) of human herpesvirus 7 is encoded by open reading frame U14 and localizes to a tegument substructure in virion particles. AB - A family of antigenically related proteins present in cells infected with human herpesvirus 7 (HHV-7), designated phosphoprotein 85 (pp85), comprises a complex of proteins, of which the 85-kDa species is phosphorylated. pp85 is a major determinant of human response to HHV-7 infection (L. Foa-Tomasi, E. Avitabile, L. Ke, and G. Campadelli-Fiume, J. Gen. Virol. 75:2719-2727, 1994; L. Foa-Tomasi, M. P. Fiorilli, E. Avitabile, and G. Campadelli-Fiume, J. Gen. Virol. 77:511-518, 1996; J. B. Black et al., Clin. Diagn. Lab. Immunol. 3:79-83, 1996). By immunoscreening of a cDNA library from HHV-7-infected cells with monoclonal antibody (MAb) 5E1, directed to the proteins of the pp85 complex, we mapped the gene encoding pp85 to the U14 open reading frame of the HHV-7 genome. A prokaryotically expressed fusion protein containing the U14 open reading frame reacted with MAb 5E1 in an immunoblot assay. A functional role for pp85 was defined by immunoelectron microscopy studies. Immunogold labeling of cryosections of HHV-7-infected cord blood mononuclear cells at high resolution localized the reactivity of MAb 5E1 to the outer surface of the virion tegument. This finding demonstrates that pp85, the product of the U14 gene, is a component of the HHV-7 tegument and suggests that the HHV-7 tegument is not a homogeneous structure but rather is composed of substructures, including an outermost layer containing pp85. The present findings, together with previously reported properties of MAb 5E1, including its ability to react with formalin-fixed paraffin-embedded samples, make this antibody a specific tool useful for etiopathogenetic studies of HHV-7 infection in humans and provide the basis for further development of pp85 into a specific recombinant diagnostic reagent. PMID- 9223463 TI - Functional management of an antiviral cytotoxic T-cell response. AB - Lymphocytic choriomeningitis virus (LCMV) is known to induce strong, polyclonal cytotoxic T-lymphocyte (CTL) responses. Using a set of variant peptides derived from the major CTL epitope of LCMV, we analyzed the functional fine specificity of the LCMV-specific CTL response. During the primary response, almost all the tested peptides were recognized. In contrast, the secondary response was purged of all minor cross-reactivities and very few peptides were significantly recognized. This study is the first demonstration of the functional maturation of a T-cell response and has important clinical and biological implications. PMID- 9223464 TI - Sequence analysis of the hepatitis C virus genome recovered from serum, liver, and peripheral blood mononuclear cells of infected chimpanzees. AB - Of 13 different strains of hepatitis C virus (HCV) in the inoculum used, only 1 persisted in human lymphocyte cell lines infected in vitro (N. Nakajima, M. Hijikata, H. Yoshikura, and Y. K. Shimizu, J. Virol. 70:3325-3329, 1996). To determine whether that particular strain (designated H1-2) has a tropism for lymphocytes in vivo, we sequenced hypervariable region 1 (HVR1) of the genome of HCV recovered from the sera, livers, and peripheral blood mononuclear cells (PBMC) of chimpanzees infected with plasma H77, the same inoculum used for the in vitro studies. In the PBMC collected from two chimpanzees during the early phase of infection, H1-2 was detected as the only or predominant HVR1 sequence. H1-2 was also detected in PBMC obtained during persistent infection from a chimpanzee that had been treated with immunosuppressants. From the livers of these chimpanzees, two to six different strains were recovered but H1-2 was not detected. Thus, H1-2 appeared to have an affinity for lymphocytes not only in vitro but also in vivo. In samples collected from a chimpanzee after 6 years of infection, however, such tissue compartmentalization of the HCV genome was not observed; a single strain became predominant in the serum, liver, and PBMC. An HCV strain capable of replicating in both the liver and PBMC probably emerged during in vivo replication and persisted. PMID- 9223465 TI - Human immunodeficiency virus type 1 (HIV-1) protein Vif inhibits the activity of HIV-1 protease in bacteria and in vitro. AB - Human immunodeficiency virus type 1 (HIV-1) Vif is required for productive infection of T lymphocytes and macrophages. Virions produced in the absence of Vif have abnormal core morphology and those produced in primary T cells carry immature core proteins and low levels of mature capsid (M. Simm, M. Shahabuddin, W. Chao, J. S. Allan, and D. J. Volsky, J. Virol. 69:4582-4586, 1995). To investigate whether Vif influences the activity of HIV-1 protease (PR), the viral enzyme which is responsible for processing Gag and Gag-Pol precursor polyproteins into mature virion components, we transformed bacteria to inducibly express truncated Gag-Pol fusion proteins and Vif. We examined the cleavage of polyproteins consisting of matrix to PR (Gag-PR), capsid to PR (CA-PR), and p6Pol to PR (p6Pol-PR) and evaluated HIV-1 protein processing at specific sites by Western blotting using antibodies against matrix, capsid, and PR proteins. We found that Vif modulates HIV-1 PR activity in bacteria mainly by preventing the release of mature MA and CA from Gag-PR, CA from CA-PR, and p6Pol from p6Pol-PR, with other cleavages being less affected. Using subconstructs of Vif, we mapped this activity to the N-terminal half of the molecule, thus identifying a new functional domain of Vif. Kinetic study of p6Pol-PR autocatalysis in the presence or absence of Vif revealed that Vif and N'Vif reduce the rate of PR-mediated proteolysis of this substrate. In an assay of in vitro proteolysis of a synthetic peptide substrate by purified recombinant PR we found that recombinant Vif and the N-terminal half of the molecule specifically inhibit PR activity at a molar ratio of the N-terminal half of Vif to PR of about 1. These results suggest a mechanism and site of action of Vif in HIV-1 replication and demonstrate novel regulation of a lentivirus PR by an autologous viral protein acting in trans. PMID- 9223466 TI - Enhanced mucosal and systemic immune responses to intestinal reovirus infection in beta2-microglobulin-deficient mice. AB - Enteric infection of mice with respiratory enteric orphan virus (reovirus) type 1, strain Lang elicits both humoral and cellular immune responses. To investigate the role of CD8+, alpha/beta T-cell receptor (TCR)+ T cells in mucosal immunity to an enteric pathogen, we examined immune responses and viral clearance following enteric reovirus infection in C57BL/6, B6129F2, and beta2-microglobulin deficient (beta2m-/-) mice. Analysis of Peyer's patch and lamina propria culture supernatants revealed a two- to threefold increase in levels of reovirus-specific immunoglobulin A in beta2m-/- mice compared to normal controls. These data corresponded to a similar increase in the frequency of virus-specific immunoglobulin A-producing cells in Peyer's patches and lamina propria and an increase in immunoglobulin G-producing cells in spleens from beta2m-/- mice compared to controls. These increased humoral immune responses were not due to a difference in B-cell populations because cell counts and flow cytometric analyses showed that beta2m-/- and control mice had similar numbers and percentages of B cells in mucosal and systemic tissues. Analysis of cytokine message by reverse transcriptase-PCR 5 and 10 days after infection revealed no difference in message level for transforming growth factor beta, gamma interferon, interleukin-4, interleukin-5, or interleukin-6 for all mouse strains. Virus tissue titers determined by plaque assay at 5 and 10 days after infection demonstrated that beta2m-/- mice cleared reovirus from the small intestines with the same efficiency as control mice. Collectively, these data suggest that CD8+, alpha/beta TCR+ T cells may regulate mucosal and systemic humoral immune responses to oral infection with reovirus. PMID- 9223467 TI - Molecular and biological characterization of a neurovirulent molecular clone of simian immunodeficiency virus. AB - To identify the molecular determinants of neurovirulence, we constructed an infectious simian immunodeficiency virus (SIV) molecular clone, SIV/17E-Fr, that contained the 3' end of a neurovirulent strain of SIV, SIV/17E-Br, derived by in vivo virus passage. SIV/17E-Fr is macrophage tropic in vitro and neurovirulent in macaques. In contrast, a molecular clone, SIV/17E-Cl, that contains the SU and a portion of the TM sequences of SIV/17E-Br is macrophage tropic but not neurovirulent. To identify the amino acids that accounted for the replication differences between SIV/17E-Fr and SIV/17E-Cl in primary macaque cells in vitro, additional infectious molecular clones were constructed. Analysis of these recombinant viruses revealed that changes in the TM portion of the envelope protein were required for the highest level of replication in primary macaque macrophages and brain cells derived from the microvessel endothelium. In addition, a full-length Nef protein is necessary for optimum virus replication in both of these cell types. Finally, viruses expressing a full-length Nef protein in conjunction with the changes in the TM had the highest specific infectivity in a sMAGI assay. Thus, changes in the TM and nef genes between SIV/17E-Cl and SIV/17E-Fr account for replication differences in vitro and correlate with replication in the central nervous system in vivo. PMID- 9223468 TI - Proteolytic processing in African swine fever virus: evidence for a new structural polyprotein, pp62. AB - We have identified an open reading frame (ORF), CP530R, within the EcoRI C' fragment of the African swine fever virus (ASFV) genome that encodes a polyprotein of 62 kDa (pp62). Antisera raised against different regions of ORF CP530R recognized a polypeptide of 62 kDa in ASFV-infected cells during the late phase of virus replication, after the onset of viral DNA synthesis. Pulse-chase experiments showed that polyprotein pp62 is posttranslationally processed to give rise to two proteins of 35 kDa (p35) and 15 kDa (p15). This proteolytic processing was found to take place at the consensus sequence Gly-Gly-X through an ordered cascade of proteolytic cleavages like that which also occurs with ASFV polyprotein pp220 (C. Simon-Mateo, G. Andres, and E. Vinuela, EMBO J. 12:2977 2987, 1993). Immunofluorescence studies showed that polyprotein pp62 is localized in the viral factories. In addition, immunoprecipitation analysis of purified virus particles showed that mature products p35 and p15 are major structural proteins. According to these results, polyprotein processing represents an essential strategy for the maturation of ASFV structural proteins. PMID- 9223469 TI - Characterization of a BHK(TK-) cell clone resistant to postattachment entry by herpes simplex virus types 1 and 2. AB - BHK(TK-) cells selected for resistance to polyethylene glycol-mediated fusion give rise to clones that are resistant to herpes simplex virus (HSV) infection. We have characterized one such clone, designated 95-19, and found that it is resistant to entry of HSV type 1 (HSV-1), HSV-2, and the related alphaherpesvirus pseudorabies virus (PRV). Single-step growth experiments show no detectable replication of multiple strains of HSV-1 and HSV-2 on 95-19 cells. Three lines of evidence suggest that these cells are resistant to postattachment entry. (i) Measurements of binding of radiolabeled virus show that heparin-sensitive binding of HSV-1 and HSV-2 to 95-19 cells is identical to binding to BHK(TK-) cells, suggesting that the block to replication occurs after attachment to heparan sulfate proteoglycan. (ii) 95-19 cells exposed to HSV-1 or HSV-2 at high multiplicity show no detectable immediate-early (IE) mRNA expression. (iii) Exposure of attached virus and cells to polyethylene glycol results in partial recovery of both IE gene expression and virus yield in single-step growth. The degrees of recovery of single-step yield and IE gene expression are similar, suggesting that the only block to single-step replication is at the point of virus entry and that these cells are deficient in some cellular factor required for efficient postattachment entry of free virus. 95-19 cells are also highly resistant to entry by cell-to-cell spread, suggesting that the same cellular factor participates in both types of entry. PMID- 9223470 TI - The temperature-sensitive (ts) phenotype of a cold-passaged (cp) live attenuated respiratory syncytial virus vaccine candidate, designated cpts530, results from a single amino acid substitution in the L protein. AB - cpts530, a candidate live-virus vaccine, is an attenuated strain of human respiratory syncytial virus (RSV). It was derived by subjecting a cold-passaged (cp) strain of RSV to a single round of chemical mutagenesis. cpts530 is a temperature-sensitive (ts) mutant that is attenuated in mice and chimpanzees, and its ts phenotype exhibits a high level of stability during replication in both species. In the present study, the complete nucleotide sequence of cpts530 RSV was determined. The five mutations known to be present in the parent cpRSV were retained in its cpts530 derivative, and one additional nucleotide change was identified at nucleotide (nt) 10060, which resulted in a phenylalanine-to-leucine change at amino acid 521 in the large polymerase (L) protein. To determine if this single amino acid substitution was indeed responsible for the ts phenotype of cpts530, it was introduced alone or in combination with the cp mutations into the full-length cDNA clone of the wild-type A2 RSV. Analysis of infectious viruses recovered from mutant cDNAs indicated that this single mutation specified complete restriction of plaque formation of recombinant cp530 in HEp-2 cell monolayer cultures at 40 degrees C, and the level of temperature sensitivity was not influenced by the presence of the five cpRSV mutations. These findings identify the phenylalanine-to-leucine change at amino acid 521 in the L protein as the mutation that specifies the ts phenotype of cpts530. Furthermore, these findings illustrate the feasibility of using the cDNA-based recovery system to analyze and construct defined attenuated vaccine viruses. PMID- 9223471 TI - The gE and gI homologs from two alphaherpesviruses have conserved and divergent neuroinvasive properties. AB - The membrane glycoproteins gE and gI are encoded by pseudorabies virus (PRV), a neurotropic, broad-host-range alphaherpesvirus of swine. PRV gE and gI are required for anterograde spread to a restricted set of retinorecipient neurons in the brain after infection of the rat retina. A related alphaherpesvirus, encoding gE and gI homologs, is called bovine herpesvirus 1.1 (BHV-1.1). BHV-1.1 is a respiratory pathogen of highly restricted host range and, in contrast to PRV, is unable to propagate in or cause disease in rodents. We have shown previously that the BHV-1.1 gE and gI proteins are capable of complementing the virulence functions of PRV gE and gI in a rodent model (A. C. Knapp and L. W. Enquist, J. Virol. 71:2731-2739, 1997). We examined the ability of the BHV-1.1 gE and gI homologs to direct circuit-specific invasion of the rat central nervous system by PRV. Both complete open reading frames were cloned into a PRV mutant lacking the PRV gE and gI genes. Recombinant viruses were analyzed for the ability to invade the rat brain after infection of the retina. Surprisingly, in a portion of the animals tested, the BHV-1.1 gE and gI proteins functioned autonomously to promote spread of PRV to a subset of retinorecipient regions of the brain. First, the presence of BHV-1.1 gI alone, but not PRV gI alone, promoted viral invasion of the optic tectum. Second, expression of BHV-1.1 gE alone facilitated PRV infection of a subset of neurons in the hippocampus not normally infected by PRV. When both BHV-1.1 proteins were expressed in a coinfection, all retinorecipient regions of the rat brain were infected. Therefore, depending on the viral source, homologs of gE and gI differentially affect spread between synaptically connected neurons in the rat. PMID- 9223472 TI - Neutralizing human monoclonal antibodies to conformational epitopes of human T cell lymphotropic virus type 1 and 2 gp46. AB - Ten human monoclonal antibodies derived from peripheral B cells of a patient with human T-cell lymphotropic virus (HTLV)-associated myelopathy are described. One monoclonal antibody recognized a linear epitope within the carboxy-terminal 43 amino acids of HTLV gp21, and two monoclonal antibodies recognized linear epitopes within HTLV type 1 (HTLV-1) gp46. The remaining seven monoclonal antibodies recognized denaturation-sensitive epitopes within HTLV-1 gp46 that were expressed on the surfaces of infected cells. Two of these antibodies also bound to viable HTLV-2 infected cells and immunoprecipitated HTLV-2 gp46. Virus neutralization was determined by syncytium inhibition assays. Eight monoclonal antibodies, including all seven that recognized denaturation-sensitive epitopes within HTLV-1 gp46, possessed significant virus neutralization activity. By competitive inhibition analysis it was determined that these antibodies recognized at least four distinct conformational epitopes within HTLV-1 gp46. These findings indicate the importance of conformational epitopes within HTLV-1 gp46 in mediating a neutralizing antibody response to HTLV infection. PMID- 9223473 TI - The effect of viral regulatory protein expression on gene delivery by human immunodeficiency virus type 1 vectors produced in stable packaging cell lines. AB - We describe the generation of stable human immunodeficiency virus type 1 (HIV-1) packaging lines that constitutively express high levels of HIV-1 structural proteins in either a Rev-dependent or a Rev-independent fashion. These cell lines were used to assess gene transfer by using an HIV-1 vector expressing the hygromycin B resistance gene and to study the effects of Rev, Tat, and Nef on the vector titer. The Rev-independent cell lines were created by using gag-pol and env expression vectors that contain the Mason-Pfizer monkey virus (MPMV) constitutive transport element (CTE). Vector titers approaching 10(4) CFU/ml were routinely obtained with these cell lines, as well as with the Rev-dependent cell lines, with HeLa-CD4 cells as targets. The presence of Nef and Tat in the producer cell each increased the vector titer 5- to 10-fold. Rev, on the other hand, was absolutely essential for gene transfer, unless the MPMV CTE was present in the vector. In that case, by using the Rev-independent cell lines for packaging, Rev could be completely eliminated from the system without a reduction in vector titer. PMID- 9223474 TI - Selection of a nonconsensus branch point is influenced by an RNA stem-loop structure and is important to confer stability to the herpes simplex virus 2 kilobase latency-associated transcript. AB - Herpes simplex virus type 1 latent infection in sensory neurons is characterized by the highly restricted transcription of viral genes. The latency-associated transcripts (LAT) family members are the only transcripts that can be identified in large amounts in latently infected cells. The most abundant LAT species is a 2 kb RNA that results from splicing of a rare primary transcript. Analysis of a LAT mutant virus (TB1) in cell culture revealed an aberrant splicing pattern and production of a stable small (0.95-kb) LAT intron. A panel of deletion constructs expressing truncated LAT in transiently transfected cells mapped the region influencing stability to the 3' end of the LAT intron. This region encompasses the branch point and a putative stable stem-loop hairpin structure immediately upstream of the splice acceptor consensus polypyrimidine tract. Mutagenic analysis of the sequence in this region confirmed our hypothesis that the stem loop structure is important for efficient splicing by influencing the selection of a nonconsensus branch point. Changes in this structure correlate with changes in branch point selection and production of an unstable 2-kb LAT. PMID- 9223476 TI - Pseudotyping human immunodeficiency virus type 1 (HIV-1) by the glycoprotein of vesicular stomatitis virus targets HIV-1 entry to an endocytic pathway and suppresses both the requirement for Nef and the sensitivity to cyclosporin A. AB - Human immunodeficiency virus type 1 (HIV-1) normally enters cells by direct fusion with the plasma membrane. In this report, HIV-1 particles capable of infecting cells through an endocytic pathway are described. Chimeric viruses composed of the HIV-1 core and the envelope glycoprotein of vesicular stomatitis virus (VSV-G) were constructed and are herein termed HIV-1(VSV) pseudotypes. HIV 1(VSV) pseudotypes were 20- to 130-fold more infectious than nonpseudotyped HIV 1. Infection by HIV-1(VSV) pseudotypes was markedly diminished by ammonium chloride and concanamycin A, a selective inhibitor of vacuolar H+ ATPases, demonstrating that these viruses require endosomal acidification to achieve productive infection. HIV-1 is thus capable of performing all of the viral functions necessary for infection when entry is targeted to an endocytic route. Maximal HIV-1 infectivity requires the presence of the viral Nef protein and the cellular protein cyclophilin A (CyPA) during virus assembly. Pseudotyping by VSV G markedly suppressed the requirement for Nef. HIV-1(VSV) particles were also resistant to inhibition by cyclosporin A; however, the deleterious effect of a gag mutation inhibiting CyPA incorporation was not relieved by VSV-G. These results suggest that Nef acts at a step of the HIV-1 life cycle that is either circumvented or facilitated by targeting virus entry to an endocytic pathway. The findings also support the hypothesis that Nef and CyPA enhance HIV-1 infectivity through independent processes and demonstrate a mechanistic difference between reduction of HIV-1 infectivity by cyclosporin A and gag mutations that decrease HIV-1 incorporation of CyPA. PMID- 9223475 TI - Human cytomegalovirus IE2 86-kilodalton protein binds p53 but does not abrogate G1 checkpoint function. AB - Physical interactions between human cytomegalovirus (HCMV) immediate-early (IE) proteins and key cell cycle regulatory proteins have been suggested as a mechanism whereby this herpesvirus modifies cellular control of proliferation. Observed similarities to interactions of other DNA virus proteins (human papillomavirus type 16 E6 and E7, simian virus 40 large T antigen, and adenovirus type 5 E1A and E1B) with cell cycle modulatory proteins such as p53 and Rb have suggested that HCMV IE proteins may likewise alter the G1-to-S phase transition. The IE2 region gene product IE86 has been shown to specifically bind p53, potentially modifying p53 G1 checkpoint function. To examine this possibility, p53-mediated G1 arrest in the presence of IE86 was assessed. Retroviral constructs were created to facilitate the stable expression of IE86 and IE72, another IE protein implicated in HCMV-mediated alteration of cell cycle progression. Western analysis and immunoprecipitation confirmed IE protein expression and binding of IE86 to p53, respectively. Chloramphenicol acetyltransferase assays examining the ability of IE86 to repress activity from the HCMV major IE promoter or activate the HCMV early promoter for the 2.2-kb class of RNAs demonstrated the functional integrity of the IE86 protein. Induction of DNA damage in normal, uninfected fibroblasts (FB) or FB expressing IE86 by actinomycin D (Act D) resulted in increased p53 levels, a predominance of the hypophosphorylated form of Rb, and increased expression of both p21(CIP1/WAF1) and mdm-2. Fluorescence-activated cell sorting revealed that both uninfected and IE86-expressing FB experienced dramatic G1 arrest following exposure to Act D. The clear demonstration of these p53-dependent responses in the presence of IE86 indicates that binding to this viral protein does not compromise the ability of p53 to elicit growth arrest following DNA damage. PMID- 9223477 TI - A viral function represses accumulation of transcripts from productive-cycle genes in mouse ganglia latently infected with herpes simplex virus. AB - Latent infections of neurons by herpes simplex virus form reservoirs of recurrent viral infections that resist cure. In latently infected neurons, viral gene expression is severely repressed; only the latency-associated transcripts (LATs) are expressed abundantly. Using sensitive reverse transcriptase PCR assays, we analyzed the effects of a deletion mutation in the LAT locus on viral gene expression in latently infected mouse trigeminal ganglia. The deletion mutation, which reduced expression of the major LATs 10(5)-fold, resulted in a approximately 5-fold increase in accumulation of transcripts from the immediate early gene encoding ICP4, an essential transactivator of viral gene expression. The LAT deletion also resulted in a >10-fold increase in the accumulation of transcripts from the early gene encoding thymidine kinase, whose expression during productive infection stringently depends on ICP4, and positively affected the correlation of the levels of these transcripts with the levels of ICP4 transcripts. We also detected transcripts antisense to ICP4 RNA, which were in substantial excess to ICP4 transcripts in ganglia latently infected with wild type virus. In contrast to its effects on productive-cycle transcripts, the LAT deletion reduced the accumulation of these antisense transcripts approximately 15 fold. Thus, a viral function associated with the LAT locus represses the accumulation of transcripts from at least two productive-cycle genes in latently infected mouse ganglia. We discuss possible mechanisms and consequences of this repression. PMID- 9223478 TI - A LAT-associated function reduces productive-cycle gene expression during acute infection of murine sensory neurons with herpes simplex virus type 1. AB - Herpes simplex virus (HSV) persists in the human population by establishing long term latent infections followed by periodic reactivation and transmission. Latent infection of sensory neurons is characterized by repression of viral productive cycle gene expression, with abundant transcription limited to a single locus that encodes the latency-associated transcripts (LATs). We have observed that LAT- deletion mutant viruses express viral productive-cycle genes in greater numbers of murine trigeminal ganglion neurons than LAT+ HSV type 1 at early times during acute infection but show reduced reactivation from latent infection. Thus, a viral function associated with the LAT region exerts an effect at an early stage of neuronal infection to reduce productive-cycle viral gene expression. These results provide the first evidence that the virus plays an active role in down regulating productive infection during acute infection of sensory neurons. The effect of down-regulation of productive-cycle gene expression during acute infection may contribute to viral evasion from the host immune responses and to reduced cytopathic effects, thereby facilitating neuronal survival and the establishment of latency. PMID- 9223479 TI - Complete sequence and genomic analysis of murine gammaherpesvirus 68. AB - Murine gammaherpesvirus 68 (gammaHV68) infects mice, thus providing a tractable small-animal model for analysis of the acute and chronic pathogenesis of gammaherpesviruses. To facilitate molecular analysis of gammaHV68 pathogenesis, we have sequenced the gammaHV68 genome. The genome contains 118,237 bp of unique sequence flanked by multiple copies of a 1,213-bp terminal repeat. The GC content of the unique portion of the genome is 46%, while the GC content of the terminal repeat is 78%. The unique portion of the genome is estimated to encode at least 80 genes and is largely colinear with the genomes of Kaposi's sarcoma herpesvirus (KSHV; also known as human herpesvirus 8), herpesvirus saimiri (HVS), and Epstein Barr virus (EBV). We detected 63 open reading frames (ORFs) homologous to HVS and KSHV ORFs and used the HVS/KSHV numbering system to designate these ORFs. gammaHV68 shares with HVS and KSHV ORFs homologous to a complement regulatory protein (ORF 4), a D-type cyclin (ORF 72), and a G-protein-coupled receptor with close homology to the interleukin-8 receptor (ORF 74). One ORF (K3) was identified in gammaHV68 as homologous to both ORFs K3 and K5 of KSHV and contains a domain found in a bovine herpesvirus 4 major immediate-early protein. We also detected 16 methionine-initiated ORFs predicted to encode proteins at least 100 amino acids in length that are unique to gammaHV68 (ORFs M1 to 14). ORF M1 has striking homology to poxvirus serpins, while ORF M11 encodes a potential homolog of Bcl-2-like molecules encoded by other gammaherpesviruses (gene 16 of HVS and KSHV and the BHRF1 gene of EBV). In addition, clustered at the left end of the unique region are eight sequences with significant homology to bacterial tRNAs. The unique region of the genome contains two internal repeats: a 40-bp repeat located between bp 26778 and 28191 in the genome and a 100-bp repeat located between bp 98981 and 101170. Analysis of the gammaHV68, HVS, EBV, and KSHV genomes demonstrated that each of these viruses have large colinear gene blocks interspersed by regions containing virus-specific ORFs. Interestingly, genes associated with EBV cell tropism, latency, and transformation are all contained within these regions encoding virus-specific genes. This finding suggests that pathogenesis-associated genes of gammaherpesviruses, including gammaHV68, may be contained in similarly positioned genome regions. The availability of the gammaHV68 genomic sequence will facilitate analysis of critical issues in gammaherpesvirus biology via integration of molecular and pathogenetic studies in a small-animal model. PMID- 9223480 TI - Both conserved region 1 (CR1) and CR2 of the human papillomavirus type 16 E7 oncogene are required for induction of epidermal hyperplasia and tumor formation in transgenic mice. AB - High-risk human papillomavirus type 16 (HPV-16) and HPV-18 are associated with the majority of human cervical carcinomas, and two viral genes, HPV E6 and E7, are commonly found to be expressed in these cancers. The presence of HPV-16 E7 is sufficient to induce epidermal hyperplasia and epithelial tumors in transgenic mice. In this study, we have performed experiments in transgenic mice to determine which domains of E7 contribute to these in vivo properties. The human keratin 14 promoter was used to direct expression of mutant E7 genes to stratified squamous epithelia in mice. The E7 mutants chosen had either an in frame deletion in the conserved region 2 (CR2) domain, which is required for binding of the retinoblastoma tumor suppressor protein (pRb) and pRb-like proteins, or an in-frame deletion in the E7 CR1 domain. The CR1 domain contributes to cellular transformation at a level other than pRb binding. Four lines of animals transgenic for an HPV-16 E7 harboring a CR1 deletion and five lines harboring a CR2 deletion were generated and were observed for overt and histological phenotypes. A detailed time course analysis was performed to monitor acute effects of wild-type versus mutant E7 on the epidermis, a site of high level expression. In the transgenic mice with the wild-type E7 gene, age dependent expression of HPV-16 E7 correlated with the severity of epidermal hyperplasia. Similar age-dependent patterns of expression of the mutant E7 genes failed to result in any phenotypes. In addition, the transgenic mice with a mutant E7 gene did not develop tumors. These experiments indicate that binding and inactivation of pRb and pRb-like proteins through the CR2 domain of E7 are necessary for induction of epidermal hyperplasia and carcinogenesis in mouse skin and also suggest a role for the CR1 domain in the induction of these phenotypes through as-yet-uncharacterized mechanisms. PMID- 9223481 TI - The 222- to 234-kilodalton latent nuclear protein (LNA) of Kaposi's sarcoma associated herpesvirus (human herpesvirus 8) is encoded by orf73 and is a component of the latency-associated nuclear antigen. AB - Kaposi's sarcoma (KS)-associated herpesvirus or human herpesvirus 8 (KSHV/HHV8) is the likely cause of KS and primary effusion lymphomas or body cavity-based lymphomas (BCBLs). A latency-associated nuclear immunofluorescence antigen (LANA) (D. H. Kedes, E. Operskalski, M. Busch, R. Kohn, J. Flood, and D. Ganem, Nat. Med. 2:918-924, 1996; S. J. Gao, L. Kingsley, M. Li, W. Zheng, C. Parravicini, J. Ziegler, R. Newton, C. R. Rinaldo, A. Saah, J. Phair, R. Detels, Y. Chang, and P. S. Moore, Nat. Med. 2:925-928, 1996) and a 222- to 234-kDa nuclear protein (LNA) (S. J. Gao, L. Kingsley, D. R. Hoover, T. J. Spira, C. R. Rinaldo, A. Saah, J. Phair, R. Detels, P. Parry, Y. Chang, and P. S. Moore, N. Engl. J. Med. 335:233 241, 1996) have previously been described in BCBL cell lines by immunofluorescence and Western blotting techniques, respectively. To identify the viral gene(s) encoding this antigen(s) we screened a cDNA library from HBL-6 cells, a B-cell lymphoma cell line persistently infected with KSHV/HHV8, with KS patient sera. One set of positive clones contained the 3' end of orf73, as well as the complete orf72 and orfK13, and another set contained the 5' end of orf73. Comparison of cDNA sequences with the KSHV/HHV8 genomic sequence revealed a splice event, occurring upstream of orf73. Immunoaffinity purified antibodies to a recombinant carboxy-terminal fragment of the orf73-encoded protein showed the characteristic speckled nuclear immunofluorescence pattern of LANA and reacted with the 222- to 234-kDa LNA on Western blots. Expression of full-length orf73 in bacteria and COS7 cells reproduced the LNA banding pattern. Immunohistochemistry on cases of nodular KS revealed that orf73/LNA is expressed in the nucleus of KS spindle cells. These findings demonstrate that orf73 encodes the 222- to 234-kDa LNA, is a component of LANA, and is expressed in KS tumor cells. PMID- 9223482 TI - Consequences of a subtle sialic acid modification on the murine polyomavirus receptor. AB - Polyomaviruses are small, nonenveloped DNA tumor viruses with restricted host ranges. Virus binding to cell surface receptors is one determinant of viral tropism. Although murine polyomavirus is among the best characterized viruses, little is known about the sialic acid-containing receptor and its interaction with viral particles. By using nonradioactive virus binding assays as recently described for the B-lymphotropic papovavirus, murine polyomavirus particles were found to bind in a saturable and noncooperative manner to 25,000 receptors per 3T6 mouse fibroblast. The virus-receptor interaction at 4 degrees C was of high affinity (Kd = 1.8 x 10(-11) M), very fast (k1 = 1.7 x 10(7) M(-1) s(-1)), and stable (half-life = 38 min). Elongation of the N-acyl side chain of sialic acid by biosynthetic modulation with synthetic precursor analogs has been shown for other polyomaviruses to influence both sialic acid-dependent binding and infection (O. T. Keppler, P. Stehling, M. Herrmann, H. Kayser, D. Grunow, W. Reutter, and M. Pawlita, J. Biol. Chem. 270:1308-1314, 1995). In 3T6 cells in which about one-third of the sialic acids were modified, infection and binding of polyomavirus particles were significantly reduced. The number of receptors per cell was decreased to 18,000, with the remaining receptors displaying the same affinity as in untreated cells. Molecular modeling studies based on the three dimensional structure of a mouse polyomavirus-sialyllactose complex recently solved by T. Stehle and coworkers (T. Stehle, Y. W. Yan, T. L. Benjamin, and S. C. Harrison, Nature 369:160-163, 1994) were performed. They suggest that the elongation of the N-acyl side chain by a single methylene group leads to steric hinderence, with the peptide backbone of a loop walling the tip of the shallow sialic acid binding groove. This collision appears to be incompatible with functional binding. The data are taken as a basis to discuss possible features of the organization and topology of the cellular receptor for mouse polyomavirus. PMID- 9223483 TI - Transduction by adeno-associated virus vectors in the rabbit airway: efficiency, persistence, and readministration. AB - The ability of recombinant adeno-associated virus (AAV) vectors to integrate into the host genome and to transduce nondividing cells makes them attractive as vehicles for gene delivery. In this study, we assessed the ability of several AAV vectors to transduce airway cells in rabbits by measuring marker gene expression. AAV vectors that transferred either a beta-galactosidase (beta-gal) or a human placental alkaline phosphatase (AP) gene were delivered to one lobe of the rabbit lung by use of a balloon catheter placed under fluoroscopic guidance. We observed vector-encoded beta-gal or AP staining almost exclusively in the epithelial and smooth muscle cells in the bronchus at the region of balloon placement. The overall efficiency of transduction in the balloon-treated bronchial epithelium was low but reached 20% in some areas. The majority of the staining was in ciliated cells but was also observed in basal cells and airway smooth muscle cells. We observed an 80-fold decrease in marker-positive epithelial cells during the 60-day period after vector infusion, whereas the number of marker-positive smooth muscle cells stayed constant. Although treatment with the topoisomerase inhibitor etoposide dramatically enhanced AAV transduction in primary airway epithelial cells in culture, treatment of rabbits did not improve transduction rates in the airway. Vector readministration failed to produce additional transduction events, which correlated with the appearance of neutralizing antibodies. These results indicate that both readministration and immune modulation will be required in the use of AAV vectors for gene therapy to the airway epithelium. PMID- 9223484 TI - Two classes of human papillomavirus type 16 E1 mutants suggest pleiotropic conformational constraints affecting E1 multimerization, E2 interaction, and interaction with cellular proteins. AB - Random mutagenesis of human papillomavirus type 16 (HPV16) E1 was used to generate E1 missense mutants defective for interaction with either hUBC9 or 16E1 BP, two cDNAs encoding proteins that have been identified by their ability to interact with HPV16 E1 in two-hybrid assays. hUBC9, the human counterpart of Saccharomyces cerevisiae UBC9, is a ubiquitin-conjugating enzyme known to be involved in cell cycle progression. 16E1-BP encodes a protein of no known function but does contain an ATPase signature motif. Eight hUBC9 or 16E1-BP interaction-defective HPV16 E1 missense mutants were identified and characterized for origin-dependent transient DNA replication, ATPase activity, and various protein-protein interaction phenotypes. Six of these mutant E1 proteins were significantly impaired for replication. Among these, two classes of replication defective HPV16 E1 missense mutants were observed. One class, represented by the S330R replication-defective mutant (containing an S-to-R change at position 330), remained competent for all protein-protein interactions tested, with the exception of hUBC9 association. Furthermore, this mutant, unlike the other replication-defective HPV16 E1 missense mutants, had a strong dominant negative replication phenotype in transient-replication assays. The other class, represented by five of the missense mutants, was defective for multiple protein protein interactions, usually including, but not limited to, the interaction defect for which each mutant was originally selected. In many cases, a single missense mutation in one region of HPV16 E1 had pleiotropic effects, even upon activities thought to be associated with other domains of HPV16 E1. This suggests that E1 proteins are not modular but may instead be composed of multiple structurally and/or functionally interdependent domains. PMID- 9223485 TI - The VP16 paradox: herpes simplex virus VP16 contains a long-range activation domain but within the natural multiprotein complex activates only from promoter proximal positions. AB - Removal of core promoter elements like the TATA box converts several regulatory upstream regions of viral and cellular genes into classical enhancers, i.e., cis regulatory elements capable of activating transcription over long distances in an orientation-independent manner. This is not the case with herpes simplex virus (HSV) immediate-early gene promoters, which are strongly induced by the viral transactivator VP16 (Vmw65, alphaTIF, ICP25) complexed with the cellular factors Oct-1 and HCF. Here we report that the VP16 complex can readily bring about strong activation from a promoter-proximal position but fails to induce transcription from a distal downstream enhancer position. This is in striking contrast to results obtained with GAL fusion proteins: in this context, the C terminal "general" activation domain of VP16 activates transcription to high levels over long distances. Thus, this paradoxical behavior suggests that the VP16 activation domain is not accessible to the transcription machinery when the VP16-Oct-1-HCF complex is bound in a remote position. Only upon specific interactions in a promoter-proximal position, perhaps with the basal transcription factors, can transcription be strongly induced. In agreement with such a proposed mechanism, VP16 proteins to which a heterologous general activation domain has been added strongly activate transcription from a downstream position. The biological role of this unexpected and sophisticated mechanism is most probably a limitation of the VP16 activity to the associated immediate-early genes, without undesired long-range effects on other viral promoters within the tightly packed HSV genome. PMID- 9223486 TI - Effect of enforced expression of human bcl-2 on Japanese encephalitis virus induced apoptosis in cultured cells. AB - Infection by Japanese encephalitis virus (JEV), a mosquito-borne flavivirus, causes acute encephalitis in humans and induces severe cytopathic effects in different types of cultured cells. This study attempted to determine whether apoptosis contributes to virus-induced cell death in a culture system by characterizing JEV lytic infection in baby hamster kidney BHK-21 cells, murine neuroblastoma N18 cells, and human neuronal progenitor NT2 cells. According to our results, the replication of JEV, and not the UV-inactivated virions per se, triggered apoptosis in these cell lines, as evidenced by nuclear condensation, DNA fragmentation ladder, and in situ end labeling of DNA strand breaks with terminal transferase (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling assay). Different strains of JEV, regardless of whether they are neurovirulent to mice, could induce apoptosis of the infected cells. In addition, enforced expression of the human protooncogene bcl-2 in BHK-21 cells, which did not influence virus production, appeared to delay the process of JEV induced apoptosis, despite the fact that most infected cells were inevitably killed after prolonged cultures. However, Bcl-2 proteins expressed in N18 cells failed to block JEV-induced apoptosis, although they did prevent Sindbis virus induced apoptosis from occurring in the same cells. This finding suggests that these two viruses may utilize similar but not identical mechanisms to kill their infected cells. The results presented here thus demonstrate that apoptosis can be a general mechanism for JEV-induced cell death and that enforced bcl-2 expression may be inadequate in protecting all cell types from JEV-induced apoptosis in cell cultures. PMID- 9223488 TI - High-efficiency incorporation of functional influenza virus glycoproteins into recombinant vesicular stomatitis viruses. AB - We derived recombinant vesicular stomatitis virus (VSV) expressing either influenza virus hemagglutinin (HA) or neuraminidase (NA) glycoproteins from extra genes inserted in the viral genome. The HA protein was expressed from a site downstream of the VSV glycoprotein (G) gene, while NA protein was expressed from a site upstream of the VSV G gene. The HA protein was expressed at lower levels than the VSV G protein, while the NA protein was expressed at higher levels, as expected from the gradient of VSV transcription that follows the gene order. The HA and NA proteins were transported to the cell surface and were functional as demonstrated by hemadsorption, hemolysis, and NA assays. Biochemical analysis showed that both HA and NA proteins were incorporated into VSV particles at high levels, although there was a preference for incorporation of the VSV G protein over either of the influenza virus proteins. Immunoelectron microscopy of the recombinants showed that the particles derived from the recombinants were mosaics carrying both the VSV G protein and the influenza virus membrane glycoproteins. These results extend earlier studies showing incorporation of the cellular glycoprotein CD4 and two other viral glycoproteins into VSV particles. Our results indicate that there is significant space in the VSV membrane that can accommodate foreign membrane proteins and that the foreign protein can represent as much as 35% of the total protein in the viral envelope. Incorporation of foreign proteins into VSV virions can, in many cases, occur passively in the absence of specific incorporation signals. PMID- 9223487 TI - VBP and RelA regulate avian leukosis virus long terminal repeat-enhanced transcription in B cells. AB - The avian leukosis virus (ALV) long terminal repeat (LTR) contains a compact transcription enhancer that is active in many cell types. A major feature of the enhancer is multiple CCAAT/enhancer element motifs that could be important for the strong transcriptional activity of this unit. The contributions of the three CCAAT/enhancer elements to LTR function were examined in B cells, as this cell type is targeted for ALV tumor induction following integration of LTR sequences next to the c-myc proto-oncogene. One CCAAT/enhancer element, termed a3, was found to be the most critical for LTR enhancement in transiently transfected B lymphoma cells, while in chicken embryo fibroblasts all three elements contributed equally to enhancement. Gel shift assays demonstrated that vitellogenin gene-binding protein (VBP), a member of the PAR subfamily of C/EBP factors, is a major component of the nuclear proteins binding to the a3 CCAAT/enhancer element. VBP activated transcription through the a3 CCAAT/enhancer element, supporting the idea that VBP is important for LTR enhancement in B cells. A member of the Rel family of proteins was also identified as a component of the a3 protein binding complex in B cells. Gel shift and immunoprecipitation assays indicated that this factor is RelA. Gel shift assays demonstrated that while RelA does not bind directly to the LTR CCAAT/enhancer elements, it does interact with VBP to potentiate VBP DNA binding activity. The synergistic interaction of VBP and RelA increased CCAAT/enhancer element-mediated transcription, indicating that both factors may be important for viral LTR regulation and also for expression of many cellular genes. PMID- 9223489 TI - The role of the pseudoknot at the 3' end of turnip yellow mosaic virus RNA in minus-strand synthesis by the viral RNA-dependent RNA polymerase. AB - The tRNA-like structure at the 3' end of turnip yellow mosaic virus (TYMV) RNA was studied in order to determine the role of this structure in the initiation of minus-strand synthesis in vitro. Deletions in the 5'-to-3' direction up to the pseudoknot structure did not result in a decrease of transcription efficiency. However, transcription efficiency was reduced twofold when a fragment of 21 nucleotides, comprising the 3'-terminal hairpin, was used as a template. tRNA(Phe) from yeast, Escherichia coli 5S rRNA, and the 3'-terminal 208 nucleotides of alfalfa mosaic virus RNA 3 could not be transcribed by the RNA dependent RNA polymerase (RdRp) of TYMV. Various mutations in the sequences of loop regions L1 and L2 or of stem region S1 of the pseudoknot were tested to further investigate the importance of the pseudoknot structure. The results were compared with those obtained in an earlier study on aminoacylation with the same mutants (R. M. W. Mans, M. H. van Steeg, P. W. G. Verlaan, C. W. A. Pleij, and L. Bosch, J. Mol. Biol. 223:221-232; 1992). Mutants which still harbor a stable pseudoknot, as proven by probing its structure, have a transcription efficiency very close to that of the wild-type virus. Disruption of the pseudoknot structure, however, gives rise to a drop in transcription efficiency to about 50%. No indications of base-specific interactions between L1, L2, or S1 of the pseudoknot and the RdRp were found. PMID- 9223490 TI - Hepatitis C virus envelope proteins bind lactoferrin. AB - Hepatitis C virus (HCV) has two envelope proteins, E1 and E2, which form a heterooligomer. During dissection of interacting regions of HCV E1 and E2, we found the presence of an interfering compound or compounds in skim milk. Here we report that human as well as bovine lactoferrin, a multifunctional immunomodulator, binds two HCV envelope proteins. As determined by far-Western blotting, the bacterially expressed E1 and E2 could bind lactoferrin in human milk directly separated or immunopurified and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The bindings of lactoferrin and HCV envelope proteins in vitro were confirmed by another method, the pull-down assay, with immunoprecipitated lactoferrin-bound protein A resin. By the same assay, mammal expressed recombinant E1 and E2 were also demonstrated to bind human lactoferrin efficiently in vitro. Direct interaction between E2 and lactoferrin was proved in vivo, since anti-human lactoferrin antibody efficiently coimmunoprecipitated with secreted and intracellular forms of the E2 protein, but not glutathione S transferase (GST), from lysates of HepG2 cells transiently cotransfected with the expression plasmids of human lactoferrin and gE2t-GST (the N-terminal two-thirds of E2 fused to GST) or GST. The N-terminal loop of lactoferrin, the region important for the antibacterial activity, has only a little role in the binding ability to HCV E2 but affected the secretion or stability of lactoferrin. Taken together, these results indicate the specific interaction between lactoferrin and HCV envelope proteins in vivo and in vitro. PMID- 9223491 TI - Identification of sequences downstream of the primer binding site that are important for efficient replication of human immunodeficiency virus type 1. AB - Reverse transcription of retroviruses is initiated from an 18-nucleotide (nt) primer binding site (PBS), located within the 5' region of viral genomic RNA, to which the host cell-derived tRNA primer is annealed and also involves viral genomic sequences outside the PBS. We constructed proviral DNA clones of human immunodeficiency virus (HIV) that had selective deletions of either a 7-nt segment found immediately downstream of the PBS or an extended nontranslated 54 nt stretch located immediately downstream of the PBS and containing the aforementioned 7-nt segment. Synthesis of minus-strand strong-stop DNA was assessed with MT-4 cells infected with viruses derived from COS-7 cells that had been transfected with these various constructs. We found that similar levels of minus-strand strong-stop DNA as well as DNA produced after template switching were expressed in MT-4 cells infected with COS-7-derived wild-type viruses or with viruses that had the 7-nt segment deleted. In contrast, significantly lower levels of viral DNA were detected in MT-4 cells after infection with viruses that had deletions of the 54-nt stretch. Furthermore, the molecular clone containing the 7-nt deletion was able to replicate with wild-type kinetics, while that containing the 54-nt deletion displayed a significantly diminished capacity in this regard. Further deletion analysis showed that a 16-nt segment at the 3' end of this 54-nt segment was largely responsible for these effects. We also conducted studies to determine levels of viral mRNA in COS-7 cells that had been transfected with equivalent amounts of DNA derived from either a wild-type HIV construct or our various deletion mutants. In the case of transfections performed with the 7-nt deletion mutant and wild-type HIV DNA, high levels of viral mRNA transcripts were detected, which was not the case for the 54 nt-deletion mutant. However, these various mRNAs possessed similar stabilities, as shown through studies in which transcript formation was arrested by treatment of cells with actinomycin D. Thus, the 54-nt segment of 5' nontranslated RNA, located downstream of the PBS, is involved in efficient expression of each of viral DNA, mRNA, and infectious virus. PMID- 9223492 TI - Immunodominant CD4+ T-cell epitope within nonstructural protein 3 in acute hepatitis C virus infection. AB - In acute hepatitis C virus infection, 50 to 70% of patients develop chronic disease. Considering the low rate of spontaneous viral clearance during chronic hepatitis C infection, the first few months of interaction between the patient's immune system and the viral population seem to be crucial in determining the outcome of infection. We previously reported the association between a strong and sustained CD4+ T-cell response to nonstructural protein 3 (NS3) of the hepatitis C virus and a self-limited course of acute hepatitis C infection. In this study, we identify an immunodominant CD4+ T-cell epitope (amino acids 1248 to 1261) that was recognized by the majority (14 of 23) of NS3-specific CD4+ T-cell clones from four of five patients with acute hepatitis C infection. This epitope can be presented to CD4+ T cells by HLA-DR4, -DR11, -DR12, -DR13, and -DR16. HLA-binding studies revealed a high binding affinity for 10 of 13 common HLA-DR alleles. Two additional CD4+ T-cell epitopes, amino acids 1388 to 1407 and amino acids 1450 to 1469, showed a very narrow pattern of binding to individual HLA-DR alleles. Our data suggest that the NS3-specific CD4+ T-cell response in acute hepatitis C infection is dominated by a single, promiscuous peptide epitope which could become a promising candidate for the development of a CD4+ T-cell vaccine. PMID- 9223494 TI - Homologous recombination occurs in a distinct retroviral subpopulation and exhibits high negative interference. AB - Homologous recombination and deletions occur during retroviral replication when reverse transcriptase switches templates. While recombination occurs solely by intermolecular template switching (between copackaged RNAs), deletions can occur by an intermolecular or an intramolecular template switch (within the same RNA). To directly compare the rates of intramolecular and intermolecular template switching, two spleen necrosis virus-based vectors were constructed. Each vector contained a 110-bp direct repeat that was previously shown to delete at a high rate. The 110-bp direct repeat was flanked by two different sets of restriction site markers. These vectors were used to form heterozygotic virions containing RNAs of each parental vector, from which recombinant viruses were generated. By analyses of the markers flanking the direct repeats in recombinant and nonrecombinant proviruses, the rates of intramolecular and intermolecular template switching were determined. The results of these analyses indicate that intramolecular template switching is much more efficient than intermolecular template switching and that direct repeat deletions occur primarily through intramolecular template switching events. These studies also indicate that retroviral recombination occurs within a distinct viral subpopulation and exhibits high negative interference, whereby the selection of one recombination event increases the probability that a second recombination event will be observed. PMID- 9223493 TI - Infection of primary cells by adeno-associated virus type 2 results in a modulation of cell cycle-regulating proteins. AB - It has been demonstrated that infection of primary human cells with adeno associated viruses (AAV) leads to a decrease in cellular proliferation and to growth arrest. We analyzed the molecular basis of this phenomenon and observed that infection with AAV type 2 (AAV2) had an effect on several factors engaged in the control of the mammalian cell cycle. In particular, all of the pRB family members, pRB, p107, and p130, which are involved in G1 cell cycle checkpoint control, were affected. After infection, a shift from hyper- to hypophosphorylated forms was observed. Cyclins A and B1, which are required for G1/S transition and progression into mitosis, respectively, were downregulated at the transcriptional level as well as at the protein level, whereas the G1 cyclins D1 and E remained unaffected. In addition, the steady-state levels of cyclin dependent kinases CDK1 and CDK2 and of transcription factor E2F-1 were diminished. Of all the factors known to be involved in phosphorylation of pRB family proteins, only the CDK inhibitor p21WAF1 exhibited a response to AAV2 infection. p21WAF1 mRNA was quickly and progressively upregulated in a p53 independent manner over at least 72 h. Consistent with the increased p21WAF1 protein levels, cyclin E- and cyclin A-dependent kinase activities declined to low levels and E2F-p130-cyclin-CDK2 complexes were disrupted. From these data, we conclude that the major effect of AAV2 infection on primary human fibroblasts appears to be upregulation of p21WAF1 gene expression and thus cell cycle arrest by the suppression of pRB family protein phosphorylation. PMID- 9223496 TI - Both T and B cells shed infectious mouse mammary tumor virus. AB - Mouse mammary tumor virus (MMTV) infected both B and T tissue culture cells and primary B and T cells in vivo after milk-borne transmission of the virus. The infected tissue culture cells processed viral proteins, and both these and primary B and T cells shed virus when cultured in vitro. Moreover, the infected B and T tissue culture cells transmitted virus to uninfected mammary gland cells in vitro. The level of infection of these different cell types in vivo was dependent on the strain of mouse, with C3H/HeN mice showing greater B-cell infection and BALB/c mice greater T-cell infection after nursing on MMTV-infected C3H/HeN mothers. Although their B cells were less infected, BALB/c mice developed tumors more rapidly than C3H/HeN mice. These results indicate that both infected T and B cells are potential carriers of MMTV in vivo. PMID- 9223495 TI - Enhancement of human immunodeficiency virus type 1 envelope-mediated fusion by a CD4-gp120 complex-specific monoclonal antibody. AB - The entry of human immunodeficiency virus type 1 (HIV-1) into cells is initiated by binding of the viral glycoprotein gp120-gp41 to its cellular receptor CD4. The gp120-CD4 complex formed at the cell surface undergoes conformational changes that may allow its association with an additional membrane component(s) and the eventual formation of the fusion complex. These conformational rearrangements are accompanied by immunological changes manifested by altered reactivity with monoclonal antibodies specific for the individual components and presentation of new epitopes unique to the postbinding complex. In order to analyze the structure and function of the gp120-CD4 complex, monoclonal antibodies were generated from splenocytes of BALB/c mice immunized with soluble CD4-gp120 (IIIB) molecules (J. M. Gershoni, G. Denisova, D. Raviv, N. I. Smorodinsky, and D. Buyaner, FASEB J. 7:1185-1187 1993). One of those monoclonal antibodies, CG10, was found to be strictly complex specific. Here we demonstrate that this monoclonal antibody can significantly enhance the fusion of CD4+ cells with effector cells expressing multiple HIV-1 envelopes. Both T-cell-line-tropic and macrophage-tropic envelope mediated cell fusion were enhanced, albeit at different optimal doses. Furthermore, infection of HeLa CD4+ (MAGI) cells by HIV-1 LAI, ELI1, and ELI2 strains was increased two- to fourfold in the presence of CG10 monoclonal antibodies, suggesting an effect on viral entry. These findings indicate the existence of a novel, conserved CD4-gp120 intermediate structure that plays an important role in HIV-1 cell fusion. PMID- 9223498 TI - Pathogenesis of simian immunodeficiency virus encephalitis: viral determinants of neurovirulence. AB - To examine the relationship between macrophage tropism and neurovirulence, macaques were inoculated with two recombinant hybrid viruses derived from the parent viruses SIVmac239, a lymphocyte-tropic, non-neurovirulent clone, and SIV/17E-Br, a macrophage-tropic, neurovirulent virus strain. The first recombinant, SIV/17E-Cl, contained the portion of the env gene that encodes the surface glycoprotein and a short segment of the transmembrane glycoprotein of SIV/17E-Br in the backbone of SIVmac239. Unlike SIVmac239, SIV/17E-Cl replicated productively in macrophages, demonstrating that sequences in the surface portion of env determine macrophage tropism. None of five macaques inoculated with SIV/17E-Cl developed simian immunodeficiency virus (SIV) encephalitis. The second recombinant, SIV/17E-Fr, which contained the entire env and nef genes and the 3' long terminal repeat of SIV/17E-Br in the SIVmac239 backbone, was also macrophage tropic. Six of nine macaques inoculated with SIV/17E-Fr developed SIV encephalitis ranging from mild to moderate in severity, indicating a significant (P = 0.031) difference in the neurovirulence of the two recombinants. In both groups of macaques, CD4+ cell counts declined gradually during infection and there was no significant difference in the rate of the decline between the two groups of macaques. This study demonstrated that macrophage tropism alone is not sufficient for the development of neurological disease. In addition, it showed that while sequences in the surface portion of the envelope gene determine macrophage tropism, additional sequences derived from the transmembrane portion of envelope and/or nef confer neurovirulence. PMID- 9223497 TI - Suppression of the phenotype of gamma(1)34.5- herpes simplex virus 1: failure of activated RNA-dependent protein kinase to shut off protein synthesis is associated with a deletion in the domain of the alpha47 gene. AB - Earlier studies have shown that infection of human cells by herpes simplex virus 1 (HSV-1) results in the activation of RNA-dependent protein kinase (PKR) but that the alpha subunit of eIF-2 is not phosphorylated and that protein synthesis is unaffected. In the absence of the viral gamma(1)34.5 gene, eIF-2alpha is phosphorylated and protein synthesis is prematurely shut off (J. Chou, J. J. Chen, M. Gross, and B. Roizman, Proc. Natl. Acad. Sci. USA 92:10516-10520, 1995). A second recent paper reported the selection of second-site suppressor mutants characterized by near-wild-type protein synthesis in cells infected with gamma(1)34.5- mutants (I. Mohr and Y. Gluzman, EMBO J. 15:4759-4766, 1996). Here, we report the properties of the spontaneous HSV-1 suppressor mutant Sup-1, which is characterized by spontaneous deletion of 503 bp encompassing the domain of the alpha47 gene and junction with the inverted repeats flanking the unique short (U(S)) sequence of the HSV-1 DNA resulting in the juxtaposition of the alpha47 promoter to the coding domain of the U(S)11 gene. This mutant does not exhibit the shutoff of protein synthesis characteristic of the gamma(1)34.5- virus. Specifically, Sup-1 in SK-N-SH human neuroblastoma cells (i) did not exhibit the function of the alpha47 gene characterized by a reduction in the transport of peptides across the endoplasmic reticulum of permealized cells consistent with the absence of alpha47 gene sequences, (ii) accumulated U(S)11 protein at levels analogous to those of the wild-type parent but the protein was made at earlier times after infection, as would be expected from a change in the promoter, and (iii) activated PKR like that of the parent, gamma(1)34.5- virus, but (iv) did not cause premature shutoff of protein synthesis and therefore was similar to the wild-type parent virus rather than the gamma(1)34.5- virus from which it was derived. We conclude that the mechanism by which Sup-1 blocks the shutoff of protein synthesis associated with phosphorylation of eIF-2alpha by the activated PKR is not readily explainable by a secondary mutation characterized by a deletion. PMID- 9223499 TI - Posttranslational processing and identification of a neutralization domain of the GP4 protein encoded by ORF4 of Lelystad virus. AB - GP4 is a minor structural glycoprotein encoded by ORF4 of Lelystad virus (LV). When it was immunoprecipitated from cell lysates and extracellular virus of CL2621 cells infected with LV, it was shown to have an apparent molecular mass of approximately 28 and 31 kDa, respectively. This difference in size occurred because its core N-glycans were modified to complex type N-glycans during the transport of the protein through the endoplasmic reticulum and Golgi compartment. A panel of 15 neutralizing monoclonal antibodies (MAbs) reacted with the native GP4 protein expressed by LV and the recombinant GP4 protein expressed in a Semliki Forest virus expression system. However, these MAbs did not react with the GP4 protein of U.S. isolate VR2332. To map the binding site of the MAbs, chimeric constructs composed of ORF4 of LV and VR2332 were generated. The reactivity of these constructs indicated that all the MAbs were directed against a region spanning amino acids 40 to 79 of the GP4 protein of LV. Six MAbs reacted with solid-phase synthetic dodecapeptides. The core of this site consists of amino acids 59 to 67 (SAAQEKISF). Comparison of the amino acid sequences of GP4 proteins from various European and North American isolates indicated that the neutralization domain spanning amino acids 40 to 79 is the most variable region of GP4. The neutralization domain of GP4, described here, is the first identified for LV. PMID- 9223501 TI - The tobacco mosaic virus RNA polymerase complex contains a plant protein related to the RNA-binding subunit of yeast eIF-3. AB - A sucrose density gradient-purified, membrane-bound tobacco mosaic virus (tomato strain L) (TMV-L) RNA polymerase containing endogenous RNA template was efficiently solubilized with sodium taurodeoxycholate. Solubilization resulted in an increase in the synthesis of positive-strand, 6.4-kb genome-length single stranded RNA (ssRNA) and a decrease in the production of 6.4-kbp double-stranded RNA (dsRNA) to levels close to the limits of detection. The solubilized TMV-L RNA polymerase was purified by chromatography on columns of DEAE-Bio-Gel and High Q. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and silver staining showed that purified RNA polymerase preparations consistently contained proteins with molecular masses of 183, 126, 56, 54, and 50 kDa, which were not found in equivalent material from healthy plants. Western blotting showed that the two largest of these proteins are the TMV-L-encoded 183- and 126-kDa replication proteins and that the 56-kDa protein is related to the 54.6-kDa GCD10 protein, the RNA-binding subunit of yeast eIF-3. The 126-, 183-, and 56-kDa proteins were coimmunoaffinity selected by antibodies against the TMV-L 126-kDa protein and by antibodies against the GCD10 protein. Antibody-linked polymerase assays showed that active TMV-L RNA polymerase bound to antibodies against the TMV-L 126-kDa protein and to antibodies against the GCD10 protein. Synthesis of genome-length ssRNA and dsRNA by a template-dependent, membrane-bound RNA polymerase was inhibited by antibodies against the GCD10 protein, and this inhibition was reversed by prior addition of GCD10 protein. PMID- 9223500 TI - Tiny T antigen: an autonomous polyomavirus T antigen amino-terminal domain. AB - Three mRNAs from the murine polyomavirus early region encode the three well characterized tumor antigens. We report the existence of a fourth alternatively spliced mRNA which encodes a fourth tumor antigen, tiny T antigen, which comprises the amino-terminal domain common to all of the T antigens but is extended by six unique amino acid residues. The amount of tiny T antigen in infected cells is small because of its short half-life. Tiny T antigen stimulates the ATPase activity of Hsc70, most likely because of its DnaJ-like motif. The common amino-terminal domain may interface with chaperone complexes to assist the T antigens in carrying out their diverse functions of replication, transcription, and transformation in the appropriate cellular compartments. PMID- 9223503 TI - A role for natural simian immunodeficiency virus and human immunodeficiency virus type 1 nef alleles in lymphocyte activation. AB - A T-lymphoid cell line termed 221 was derived from a rhesus monkey infected with herpesvirus saimiri. Growth of 221 cells was dependent on the addition of interleukin-2 (IL-2) to the culture medium. In the absence of IL-2, 221 cells arrested in G0-G1 but did not die. Simian immunodeficiency virus (SIV) replicated efficiently in IL-2-stimulated 221 cells whether or not the nef gene was present. In the absence of IL-2, nef-containing SIV replicated 8 to 100 times more efficiently in 221 cells than did the same virus lacking nef. nef-containing virus preferentially stimulated the production of IL-2 from 221 cells. HIV-1 nef and v-ras genes, but not the c-ras gene, were shown to substitute functionally for SIV nef when tested as recombinant viruses in this assay system. These results demonstrate a role for natural nef in causing lymphoid cell activation, and they provide a system for delineating the biochemical mechanisms responsible for this activation. PMID- 9223502 TI - Glycoprotein D of herpes simplex virus (HSV) binds directly to HVEM, a member of the tumor necrosis factor receptor superfamily and a mediator of HSV entry. AB - Glycoprotein D (gD) is a structural component of the herpes simplex virus (HSV) envelope which is essential for virus entry into host cells. Chinese hamster ovary (CHO-K1) cells are one of the few cell types which are nonpermissive for the entry of many HSV strains. However, when these cells are transformed with the gene for the herpesvirus entry mediator (HVEM), the resulting cells, CHO-HVEM12, are permissive for many HSV strains, such as HSV-1(KOS). By virtue of its four cysteine-rich pseudorepeats, HVEM is a member of the tumor necrosis factor receptor superfamily of proteins. Recombinant forms of gD and HVEM, gD-1(306t) and HVEM(200t), respectively, were used to demonstrate a specific physical interaction between these two proteins. This interaction was dependent on native gD conformation but independent of its N-linked oligosaccharides, as expected from previous structure-function studies. Recombinant forms of gD derived from HSV-1(KOS)rid1 and HSV-1(ANG) did not bind to HVEM(200t), explaining the inability of these viruses to infect CHO-HVEM12 cells. A variant gD protein, gD 1(delta290-299t), showed enhanced binding to HVEM(200t) relative to the binding of gD-1(306t). Competition studies showed that gD-1(delta290-299t) and gD-1(306t) bound to the same region of HVEM(200t), suggesting that the differences in binding to HVEM are due to differences in affinity. These differences were also reflected in the ability of gD-1(delta290-299t) but not gD-1(306t) to block HSV type 1 infection of CHO-HVEM12 cells. By gel filtration chromatography, the complex between gD-1(delta290-299t) and HVEM(200t) had a molecular mass of 113 kDa and a molar ratio of 1:2. We conclude that HVEM interacts directly with gD, suggesting that HVEM is a receptor for virion gD and that the interaction between these proteins is a step in HSV entry into HVEM-expressing cells. PMID- 9223505 TI - Amino acid changes in the Sindbis virus E2 glycoprotein that increase neurovirulence improve entry into neuroblastoma cells. AB - Sindbis virus (SV) is an alphavirus that causes encephalitis in mice and results in age-dependent mortality. The outcome is dependent on the virus strain. Residues at 55 and 172 in the E2 glycoprotein determine the neurovirulence for mice of different ages and the efficiency of replication in the nervous system and neuronal cells. To determine the effects of these two residues on the initial steps in replication, we studied viruses with a histidine or glutamine at E2 position 55 and a glycine or an arginine at position 172, E2[H55G172], E2[Q55G172], E2[H55R172], and E2[Q55R172]. The production of virus was detected earlier for viruses with a histidine at E2 position 55 in BHK-21 cells (4 to 6 versus 6 to 8 h) and for E2[H55G172] in N18 cells (6 versus 8 to 10 h). As shown previously, viruses with a glycine at E2 position 172 bound more efficiently to N18 cells and a histidine at E2 position 55 further improved binding only slightly. Viruses with E2[H55] exhibited more rapid internalization and degradation of viral proteins in both BHK-21 and N18 cells. Incubation of E2[H55G172] and E2[Q55G172] at various pHs and temperatures did not reveal differences in virion stability. These data suggest that the amino acids at E2 positions 172 and 55 affect both adsorption and penetration of SV and that these early steps in the replicative pathway contribute to increased neurovirulence. PMID- 9223504 TI - A single amino acid change in the E2 glycoprotein of Sindbis virus confers neurovirulence by altering an early step of virus replication. AB - Amino acid changes in the envelope glycoproteins of Sindbis virus have been linked to neurovirulence; however, the molecular mechanisms by which these amino acid changes alter neurovirulence are not known. Recombinant-virus studies have mapped an important determinant of neurovirulence in adult mice to a single amino acid change, glutamine to histidine, at position 55 of the E2 glycoprotein (P. C. Tucker, E. G. Strauss, R. J. Kuhn, J. H. Strauss, and D. E. Griffin, J. Virol. 67:4605-4610, 1993). To investigate how histidine confers neurovirulence, we examined the various stages of the virus life cycle in neural (N18) and nonneural (BHK) cells. In BHK cells, recombinant viruses 633 (E255Q) and TE (E255H) replicated similarly. In contrast, in N18 neuroblastoma cells, TE established infection more efficiently, replicated faster, and achieved higher rates of virus release than did 633. Viral structural protein synthesis was similar in 633- and TE-infected BHK cells, while in N18 cells, structural protein synthesis was detected only in TE-infected cells at 6 h and remained higher for at least 16 h postinfection. Viral RNA synthesis was initiated more rapidly and was up to fivefold greater in TE- versus 633-infected N18 cells. Taken together with other data demonstrating minimal effects on virus binding and entry (P. C. Tucker, S. H. Lee, N. Bui, D. Martinie, and D. E. Griffin, J. Virol. 71:6106-6112, 1997), these data suggest that E2 position 55 plays an important role at early stages of infection of neural cells, thereby facilitating neurovirulence. PMID- 9223507 TI - Coinfection of wild ducks by influenza A viruses: distribution patterns and biological significance. AB - Coinfection of wild birds by influenza A viruses is thought to be an important mechanism for the diversification of viral phenotypes by generation of reassortants. However, it is not known whether coinfection is a random event or follows discernible patterns with biological significance. In the present study, conducted with viruses collected throughout 15 years from a wild-duck population in Alberta, Canada, we identified three discrete distributions of coinfections. In about one-third of the events, which involved subtypes of viruses that appear to be maintained in this duck reservoir, coinfection occurred at rates either close to or significantly lower than one would predict from rates of single-virus infection. Apparently, the better adapted an influenza A virus is to an avian population, the greater is its ability to prevent coinfections. Conversely, poorly adapted, nonmaintained viruses were significantly overrepresented as coinfectants. Rarely encountered subtypes appear to represent viruses whose chances of successfully infiltrating avian reservoirs are increased by coinfection. Mallards (Anas platyrhynchos) and pintails (A. acuta) were significantly more likely to be infected by a single influenza A virus than were the other species sampled, but no species was significantly more likely to be coinfected. These observations provide the first evidence of nonrandom coinfection of wild birds by influenza A viruses, suggesting that reassortment of these viruses in a natural population does not occur randomly. These results suggest that even though infections may occur in a species, all subtypes are not maintained by all avian species. They also suggest that specific influenza A virus subtypes are differentially adapted to different avian hosts and that the fact that a particular subtype is isolated from a particular avian species does not mean that the virus is maintained by that species. PMID- 9223506 TI - Transcription factor binding sites downstream of the human immunodeficiency virus type 1 transcription start site are important for virus infectivity. AB - When transcriptionally active, the human immunodeficiency virus (HIV) promoter contains a nucleosome-free region encompassing both the promoter/enhancer region and a large region (255 nucleotides [nt]) downstream of the transcription start site. We have previously identified new binding sites for transcription factors downstream of the transcription start site (nt 465 to 720): three AP-1 sites (I, II, and III), an AP3-like motif (AP3-L), a downstream binding factor (DBF) site, and juxtaposed Sp1 sites. Here, we show that the DBF site is an interferon responsive factor (IRF) binding site and that the AP3-L motif binds the T-cell specific factor NF-AT. Mutations that abolish the binding of each factor to its cognate site are introduced in an infectious HIV-1 molecular clone to study their effect on HIV-1 transcription and replication. Individual mutation of the DBF or AP3-L site as well as the double mutation AP-1(III)/AP3-L did not affect HIV-1 replication compared to that of the wild-type virus. In contrast, proviruses carrying mutations in the Sp1 sites were totally defective in terms of replication. Virus production occurred with slightly delayed kinetics for viruses containing combined mutations in the AP-1(III), AP3-L, and DBF sites and in the AP3-L and DBF-sites, whereas viruses mutated in the AP-1(I,II,III) and AP3-L sites and in the AP-1(I,II,III), AP3-L, and DBF sites exhibited a severely defective replicative phenotype. No RNA-packaging defect could be measured for any of the mutant viruses as determined by quantification of their HIV genomic RNA. Measurement of the transcriptional activity of the HIV-1 promoter after transient transfection of the HIV-1 provirus DNA or of long terminal repeat luciferase constructs showed a positive correlation between the transcriptional and the replication defects for most mutants. PMID- 9223508 TI - Sp1 binds to the precise locus of end processing within the terminal repeats of Epstein-Barr virus DNA. AB - Interconversion between the linear genome of Epstein-Barr virus (EBV) present in virions and intracellular circular EBV DNA is a novel DNA recombination process. A previously characterized DNA binding activity called terminal repeat or tandem repeat binding protein (TRBP) was found to recognize several G-rich recombinogenic sequences in the EBV genome and in cellular DNA. TRBP was also found to be an autoantigen recognized by sera from certain patients with undifferentiated connective-tissue disorders. Here the transcription factor Sp1 has been identified as a component of TRBP and has been shown to be an autoantigen. Sp1 bound to recombination junctions of EBV DNA, such as those in the terminal repeats and in the large internal repeats, as well as to recombinogenic regions of cellular DNA, such as variable-number tandem repeats and switch regions of the immunoglobulin genes. We defined the ends of the linear EBV genome present in virions and showed that Sp1 binds to the sequence (GGGGTGGGGCATGGG) within EBV terminal repeats at the precise locus of interconversion of linear and circular viral DNA. Sp1 may be involved in DNA recombination. PMID- 9223509 TI - Artificial mutations and natural variations in the CD46 molecules from human and monkey cells define regions important for measles virus binding. AB - CD46 was previously shown to be a primate-specific receptor for the Edmonston strain of measles virus. This receptor consists of four short consensus regions (SCR1 to SCR4) which normally function in complement regulation. Measles virus has recently been shown to interact with SCR1 and SCR2. In this study, receptors on different types of monkey erythrocytes were employed as "natural mutant proteins" to further define the virus binding regions of CD46. Erythrocytes from African green monkeys and rhesus macaques hemagglutinate in the presence of measles virus, while baboon erythrocytes were the least efficient of the Old World monkey cells used in these assays. Subsequent studies demonstrated that the SCR2 domain of baboon CD46 contained an Arg-to-Gln mutation at amino acid position 103 which accounted for reduced hemagglutination activity. Surprisingly, none of the New World monkey erythrocytes hemagglutinated in the presence of virus. Sequencing of cDNAs derived from the lymphocytes of these New World monkeys and analysis of their erythrocytes with SCR1-specific polyclonal antibodies indicated that the SCR1 domain was deleted in these cells. Additional experiments, which used 35 different site-specific mutations inserted into CD46, were performed to complement the preceding studies. The effects of these artificial mutations were documented with a convenient binding assay using insect cells expressing the measles virus hemagglutinin. Mutations which mimicked the change found in baboon CD46 or another which deleted the SCR2 glycosylation site reduced binding substantially. Another mutation which altered GluArg to AlaAla at positions 58 and 59, totally abolished binding. Finally, the epitopes for two monoclonal antibodies which inhibit measles virus attachment were mapped to the same regions implicated by mutagenesis. PMID- 9223510 TI - Sequence and structure alignment of Paramyxoviridae attachment proteins and discovery of enzymatic activity for a morbillivirus hemagglutinin. AB - On the basis of the conservation of neuraminidase (N) active-site residues in influenza virus N and paramyxovirus hemagglutinin-neuraminidase (HN), it has been suggested that the three-dimensional (3D) structures of the globular heads of the two proteins are broadly similar. In this study, details of this structural similarity are worked out. Detailed multiple sequence alignment of paramyxovirus HN proteins and influenza virus N proteins was based on the schematic representation of the previously proposed structural similarity. This multiple sequence alignment of paramyxovirus HN proteins was used as an intermediate to align the morbillivirus hemagglutinin (H) proteins with neuraminidase. Hypothetical 3D structures were built for paramyxovirus HN and morbillivirus H, based on homology modelling. The locations of insertions and deletions, glycosylation sites, active-site residues, and disulfide bridges agree with the proposed 3D structure of HN and H of the Paramyxoviridae. Moreover, details of the modelled H protein predict previously undescribed enzymatic activity. This prediction was confirmed for rinderpest virus and peste des petits ruminants virus. The enzymatic activity was highly substrate specific, because sialic acid was released only from crude mucins isolated from bovine submaxillary glands. The enzymatic activity may indicate a general infection mechanism for respiratory viruses, and the active site may prove to be a new target for antiviral compounds. PMID- 9223511 TI - Capillary endothelial cell tropism of PVC-211 murine leukemia virus and its application for gene transduction. AB - PVC-211 murine leukemia virus (MuLV) causes neurodegenerative disease following inoculation of neonatal, but not adult, mice and rats. It was previously shown that tropism for brain capillary endothelial cells (CEC) was a determinant of the viral neuropathogenicity. In this study, we demonstrate that host age-dependent replication of PVC-211 MuLV in vivo occurs in CEC in the brain as well as in other organs, such as the liver, kidney, and heart. In contrast, primary explant cultures of CEC derived from brains and livers of adult and neonatal rats could be infected by PVC-211 MuLV, suggesting that the age-dependent susceptibility was abrogated in vitro. Although CEC were generally less susceptible to MuLV-mediated gene transduction than fibroblasts, treatment of CEC with 2-deoxyglucose followed by inoculation of a PVC-211 MuLV-pseudotyped vector in the absence of heparin improved the transduction efficiency. These observations support the possibility that PVC-211 MuLV may be useful for establishing models of CEC gene transduction. PMID- 9223512 TI - Sensitization of rhabdo-, lenti-, and spumaviruses to human serum by galactosyl(alpha1-3)galactosylation. AB - Vesicular stomatitis virus, human immunodeficiency virus type 2, and human foamy virus, which were produced by cell lines expressing galactosyl(alpha1 3)galactosyl (alphaGal) sugars, were found to be less stable in human serum than those from alphaGal-negative cells, indicating that galactosyl(alpha1 3)galactosylation sensitizes these viruses as well as mammalian type C oncoviruses (Rother et al., J. Exp. Med. 182:1345-1355, 1995; Takeuchi et al., Nature (London) 379:85-88, 1996) to complement killing via natural anti-alphaGal antibodies. Thus, virus killing mediated by anti-alphaGal antibodies may play a role as a barrier to animal-to-human infection of various enveloped viruses. Virus vectors for human in vivo gene therapy based on the viruses mentioned above should be produced from alphaGal-negative cells. PMID- 9223513 TI - Specific infection and destruction of dopaminergic neurons in the substantia nigra by Theiler's virus. AB - Theiler's murine encephalomyelitis virus was stereotaxically inoculated unilaterally into the substantia nigra of the mouse brain. Virus specifically infected tyrosine hydroxylase-positive neurons and spread rostrocaudally throughout this subpopulation of neurons, resulting in impaired function and degeneration of the substantia nigra. The spread of the virus to other areas of the brain was minimal and rare. PMID- 9223514 TI - Generation of coronavirus spike deletion variants by high-frequency recombination at regions of predicted RNA secondary structure. AB - Coronavirus RNA evolves in the central nervous systems (CNS) of mice during persistent infection. This evolution can be monitored by detection of a viral quasispecies of spike deletion variants (SDVs) (C. L. Rowe, S. C. Baker, M. J. Nathan, and J. O. Fleming, J. Virol. 71:2959-2969, 1997). We and others have found that the deletions cluster in the region from 1,200 to 1,800 nucleotides from the 5' end of the spike gene sequence, termed the "hypervariable" region. To address how SDVs might arise, we generated the predicted folding structures of the positive- and negative-strand senses of the entire 4,139-nt spike RNA sequence. We found that a prominent, isolated stem-loop structure is coincident with the hypervariable region in each structure. To determine if this predicted stem-loop is a "hot spot" for RNA recombination, we assessed whether this region of the spike is more frequently deleted than three other selected regions of the spike sequence in a population of viral sequences isolated from the CNS of acutely and persistently infected mice. Using differential colony hybridization of cloned spike reverse transcription-PCR products, we detected SDVs in which the hot spot was deleted but did not detect SDVs in which other regions of the spike sequence were exclusively deleted. Furthermore, sequence analysis and mapping of the crossover sites of 25 distinct patterns of SDVs showed that the majority of crossover sites clustered to two regions at the base of the isolated stem-loop, which we designated as high-frequency recombination sites 1 and 2. Interestingly, the majority of the left and right crossover sites of the SDVs were directly across from or proximal to one another, suggesting that these SDVs are likely generated by intramolecular recombination. Overall, our results are consistent with there being an important role for the spike RNA secondary structure as a contributing factor in the generation of SDVs during persistent infection. PMID- 9223515 TI - Truncation of the C-terminal acidic transcriptional activation domain of herpes simplex virus VP16 produces a phenotype similar to that of the in1814 linker insertion mutation. AB - We examined the phenotype of a herpes simplex virus (HSV) type 1 mutant (V422) in which the C-terminal acidic activation domain of the virion transactivator VP16 is truncated at residue 422. The efficiency of plaque formation by V422 on Vero cells was boosted by approximately 100-fold by including hexamethylene bis acetimide (HMBA) in the growth medium, as previously observed with the in1814 VP16 linker insertion mutant isolated by Preston and colleagues. V422 displayed severely reduced levels of the immediate-early transcripts encoding ICP0 and ICP4 during infection in the presence of cycloheximide, and this defect was partially overcome by the addition of HMBA. The defect in plaque formation exhibited by V422 and in 1814 was efficiently complemented in U2OS osteosarcoma cells, which had previously been shown to complement ICP0 null mutations. Taken in combination, these data confirm the key role of VP16 in triggering the onset of the HSV lytic cycle. PMID- 9223517 TI - In vitro analysis of the E1A-homologous sequences of RIZ. AB - The RIZ (G3B/MTB-Zf) gene was first isolated based on its ability to bind to the retinoblastoma protein (Rb). An acidic, approximately 100-amino-acid region around the Rb-binding motif of RIZ has structural and antigenic similarity to the conserved sequences of the E1A viral oncogene. We show here that this region interacts specifically with the E1A-binding domain of Rb. This interaction could be disrupted by E1A or by a peptide of RIZ homologous to the CR2 motif of E1A which is involved in binding to Rb family proteins. Also like E1A, RIZ can form a ternary complex with Rb and E2F1. Despite this similarity to E1A, however, RIZ could not bind to the Rb family proteins p107 and p130 in vitro. The data show that the RIZ CR2 motif can mediate differential binding to Rb family proteins. We also mapped the shared antigenic determinant between RIZ and E1A to a conserved sequence, designated CE1, which is located in the C terminus of E1A. Unlike that of ETA, the CE1 motif of RIZ is located next to the CR2 motif. Despite this proximity, CE1 and CR2 appear to act independently. The data show similarities as well as differences between the homologous sequences of RIZ and E1A and contribute to an understanding of the biochemistry of these proteins. PMID- 9223516 TI - Interaction of the UV-damaged DNA-binding protein with hepatitis B virus X protein is conserved among mammalian hepadnaviruses and restricted to transactivation-proficient X-insertion mutants. AB - We carried out a comparative analysis of several proposed host protein partners of the human hepatitis B virus X protein (HBx) using both the GAL4- and the LexA based yeast two-hybrid system. We showed that the interaction of HBx with the UV damaged DNA-binding protein (UVDDB) is positive in both yeast systems, detectable in cotransfected human cells, conserved by rodent hepadnavirus X proteins (known to transactivate in human cells), and tightly correlated with the transactivation proficiency of X-insertion mutants. Taken together, our results strongly suggest that UVDDB is involved in X-mediated transactivation. PMID- 9223518 TI - Integrin alpha5beta1-mediated adenovirus infection is enhanced by the integrin activating antibody TS2/16. AB - Adenovirus internalization generally has been accepted to involve an interaction of the adenoviral penton base protein with alpha(v)beta3 and alpha(v)beta5 cell surface integrins. In this study we show that exposure of a panel of melanoma cells to the beta1-activating antibody TS2/16 rendered such cells more susceptible to adenovirus infection. This increase in adenoviral infectivity paralleled effects on cell adhesion, and both these characteristics were mediated, in part, by the alpha5beta1 integrin. These observations suggest that alpha5beta1 may act as an alternative adenovirus receptor and that integrin activating strategies may improve the efficacy of recombinant adenoviruses as vectors for gene therapy. PMID- 9223519 TI - Probing the substrate specificity of hepatitis C virus NS3 serine protease by using synthetic peptides. AB - We probed the substrate specificity of a recombinant noncovalent complex of the full-length hepatitis C virus (HCV) NS3 serine protease and NS4A cofactor, using a series of small synthetic peptides derived from the three trans-cleavage sites of the HCV nonstructural protein sequence. We observed a distinct cleavage site preference exhibited by the enzyme complex. The values of the turnover number (k(cat)) for the most efficient NS4A/4B, 4B/5A, and 5A/5B peptide substrates were 1.6, 11, and 8 min(-1), respectively, and the values for the corresponding Michaelis-Menten constants (Km) were 280, 160, and 16 microM, providing catalytic efficiency values (k(cat)/Km) of 92, 1,130, and 8,300 M(-1) s(-1). An alanine scanning study for an NS5A/5B substrate (P6P4') revealed that P1 Cys and P3 Val were critical. Finally, substitutions at the scissile P1 Cys residue by homocysteine (Hcy), S-methylcysteine (Mcy), Ala, S-ethylcysteine (Ecy), Thr, Met, D-Cys, Ser, and penicillamine (Pen) produced progressively less efficient substrates, revealing a stringent stereochemical requirement for a Cys residue at this position. PMID- 9223520 TI - Poliovirus protein 2BC increases cytosolic free calcium concentrations. AB - Poliovirus-infected cells undergo an increase in cytoplasmic calcium concentrations from the 4th h postinfection. The protein responsible for this effect was identified by the expression of different poliovirus nonstructural proteins in HeLa cells by using a recombinant vaccinia virus system. Synthesis of protein 2BC enhances cytoplasmic calcium concentrations in a manner similar to that observed in poliovirus-infected cells. To identify the regions in 2BC involved in modifying cytoplasmic calcium levels, several 2BC variants were generated. Regions present in both 2B and 2C are necessary to augment cellular free calcium levels. Therefore, in addition to inducing proliferation of membranous vesicles, poliovirus protein 2BC also alters cellular calcium homeostasis. PMID- 9223521 TI - Psi- vectors: murine leukemia virus-based self-inactivating and self-activating retroviral vectors. AB - We have developed murine leukemia virus (MLV)-based self-inactivating and self activating vectors to show that the previously demonstrated high-frequency direct repeat deletions are not unique to spleen necrosis virus (SNV) or the neomycin drug resistance gene. Retroviral vectors pKD-HTTK and pKD-HTpTK containing direct repeats composed of segments of the herpes simplex virus type 1 thymidine kinase (HTK) gene were constructed; in pKD-HTpTK, the direct repeat flanked the MLV packaging signal. The generation of hypoxanthine-aminopterin-thymidine-resistant colonies after one cycle of retroviral replication demonstrated functional reconstitution of the HTK gene. Quantitative Southern analysis indicated that direct repeat deletions occurred in 57 and 91% of the KD-HTTK and KD-HTpTK proviruses, respectively. These results demonstrate that (i) deletion of direct repeats occurs at similar high frequencies in SNV and MLV vectors, (ii) MLV psi can be efficiently deleted by using direct repeats, (iii) suicide genes can be functionally reconstituted during reverse transcription, and (iv) the psi region may be a hot spot for reverse transcriptase template switching events. PMID- 9223522 TI - Human immunodeficiency virus type 1 nucleocapsid protein specifically stimulates Mg2+-dependent DNA integration in vitro. AB - The integrase (IN) protein of the human immunodeficiency virus mediates integration of the viral DNA into the cellular genome. In vitro, this reaction can be mimicked by using purified recombinant IN and model DNA substrates. IN mediates two reactions: an endonucleolytic cleavage at each 3' end of the proviral DNA (terminal cleavage) and the joining of the linear viral DNA to 5' phosphates in the target DNA (strand transfer). Previous investigators have shown that purified IN requires Mn2+ or Mg2+ to promote strand transfer in vitro, although Mg2+ is the likely metal cofactor in vivo. IN activity in the presence of Mg2+ in vitro requires high IN concentrations and low concentrations of salt. Here, we show that the viral nucleocapsid protein NCp7 allows efficient IN mediated strand transfer in the presence of Mg2+ at low enzyme concentrations. This potentiating effect appears to be unique to NCp7, as other small DNA-binding proteins, while capable of stimulating integration in the presence of Mn2+, all failed to stimulate strand transfer in the presence of Mg2+. PMID- 9223523 TI - Upregulation of human immunodeficiency virus (HIV) replication by CD4 cross linking in peripheral blood mononuclear cells of HIV-infected adults. AB - This study was conducted with peripheral blood mononuclear cells from 67 human immunodeficiency virus (HIV)-infected adults. It supports the hypothesis that cross-linking of CD4 molecules by HIV gp120 can result in HIV upregulation and spread of infection. Underlying mechanisms include activation of latent infection by factors in addition to, or other than, tumor necrosis factor alpha. PMID- 9223524 TI - Transcriptional strategy of closteroviruses: mapping the 5' termini of the citrus tristeza virus subgenomic RNAs. AB - Citrus tristeza virus (CTV) induces formation of a nested set of at least nine 3' coterminal subgenomic RNAs (sgRNAs) in infected tissue. The organization and expression of the 19,296-nucleotide (nt) CTV genome resembles that of coronaviruses, with polyprotein processing, translational frameshifting, and multiple sgRNA formation, but phylogenetically the CTV polymerase, like polymerases of other closteroviruses, belongs to the Sindbis virus-like lineage of RNA virus polymerases. Both positive-strand RNA virus supergroups, coronaviruses and Sindbis-like viruses, utilize different mechanisms of transcription. To address the mechanism of CTV transcription, 5' termini for the two most abundant sgRNAs, 1.5 and 0.9 kb, respectively, were mapped by runoff reverse transcription. The two sgRNAs were demonstrated to have 48- and 38-nt 5' untranslated regions (5'-UTRs), respectively. The 5'-UTR for the 1.5-kb RNA was cloned, sequenced, and demonstrated to be colinear with the 48-nt genomic sequence upstream of the initiator codon of the respective open reading frame 10, i.e., to be of continuous template origin. The data obtained suggest that the sgRNA transcription of CTV is dissimilar from the coronavirus transcription and consistent with the transcriptional mechanism of other Sindbis-like viruses. Thus, the Sindbis virus-like mechanism of transcription of the positive-strand RNA genomes might be successfully utilized by the closterovirus genome of up to 19.3 kb with multiple sgRNAs. PMID- 9223525 TI - RNAs from genetically distinct retroviruses can copackage and exchange genetic information in vivo. AB - Sequence analysis suggests that ancient recombination events may have occurred between genetically distinct retroviruses. An experimental system was utilized to explore the genetic interaction between different viruses. Moloney murine sarcoma virus and spleen necrosis virus are type C retroviruses that belong to different subgenera. With vectors containing packaging signals from these two viruses, DNA proviruses containing genetic information from both RNAs can be generated. This is the first experimental evidence to indicate that RNA from different retroviruses can copackage and exchange genetic information. PMID- 9223526 TI - Requirement of poly(rC) binding protein 2 for translation of poliovirus RNA. AB - Poly(rC) binding protein 2 (PCBP2) is one of several cellular proteins that interact specifically with a major stem-loop domain in the poliovirus internal ribosome entry site. HeLa cell extracts subjected to stem-loop IV RNA affinity chromatography were depleted of all detectable PCBP2. Such extracts were unable to efficiently translate poliovirus RNA, although extracts recovered from control columns of matrix unlinked to RNA retained full translation activity. Both translation and production of infectious progeny virus were restored in the PCBP2 depleted extracts by addition of recombinant PCBP2, but not by PCBP1, which is a closely related member of the protein family. The data show that PCBP2 is an essential factor, which is required for efficient translation of poliovirus RNA in HeLa cells. PMID- 9223527 TI - Characterization of a human papillomavirus type 16 variant-dependent neutralizing epitope. AB - We have determined that three type-specific and conformationally dependent monoclonal antibodies, H16.E70, H16.U4, and H16.V5, neutralize pseudotype human papillomavirus type 16 (HPV16) virions in vitro. H16.U4 and H16.V5 neutralized pseudotype virions derived from the German HPV16 variant 114K and the Zairian variant Z-1194 with equal efficiency. In contrast, neutralization of Z-1194 pseudotype virions by H16.E70 was two orders of magnitude weaker than neutralization of 114K pseudotype virions. This difference correlated with enzyme linked immunosorbent assay reactivity of H16.E70 to L1 virus-like particles of the two variants. A substitution at residue 282 of L1 was responsible for this differential reactivity, suggesting that this residue constitutes part of the H16.E70 epitope. PMID- 9223528 TI - Molecular characterization and phylogenetic analyses of a new, highly divergent simian T-cell lymphotropic virus type 1 (STLV-1marc1) in Macaca arctoides. AB - A serological survey of a captive colony of Asian monkeys indicated that six Macaca arctoides had antibodies to human T-cell leukemia/lymphotropic viruses (HTLV). Over a 4-year interval, sera from these animals continued to exhibit a peculiar Western blot (WB) pattern resembling an HTLV-2 pattern (p24gag reactivity of equal or greater intensity than that of p19gag and a strong reactivity to recombinant gp21) but also exhibiting, in five of six cases, a reactivity against MTA-1, an HTLV-1 gp46 peptide. PCR experiments on DNA extracted from peripheral blood mononuclear cells using HTLV-1- or HTLV-2 specific long terminal repeat, gag, pol, env, and tax primers yielded negative results. However, highly conserved primers successfully amplified three different gene segments of env, tax, and env-tax. The results of comparative sequence analysis demonstrated that STLV-1marc1 was not closely related to any known STLV 1 strain, was the most divergent strain of the HTLV-1-STLV-1 group, and lacked the ATG initiation codons corresponding to the p12 and p13 proteins of HTLV-1. Phylogenetic analyses incorporating representative strains of all known HTLV-STLV clades consistently depicted STLV-1marc1 within the HTLV-1-STLV-1 type 1 lineage, but it probably diverged early, since its position is clearly different from all known viral strains of this group and it had a bootstrap resampling value of 100%. Genetic distance estimates between STLV-1marc1 and all other type 1 viruses were of the same order of magnitude as those between STLV-2PanP and all other type 2 viruses. In light of the recent demonstration of interspecies transmission of some STLV-1 strains, our results suggest the existence in Asia of HTLV-1 strains related to this new divergent STLV-1marc1 strain, which may be derived from a common ancestor early in the evolution of the type 1 viruses and could be therefore considered a prototype of a new HTLV-STLV clade. PMID- 9223530 TI - A point mutation abolishes the helicase but not the nucleoside triphosphatase activity of hepatitis C virus NS3 protein. AB - The NS3 protein of hepatitis C virus contains a bipartite structure consisting of an N-terminal serine protease and a C-terminal DEAD box helicase. We show that the C-terminal domain has ATPase and panhelicase activities. The integrity of the helicase function is dependent on the conserved DEAD motif and can be abolished by a His-Ala point mutation, leaving a fully functional nucleoside triphosphatase. PMID- 9223529 TI - Type 1 and type 2 cytokine gene expression by viral gp135 surface protein activated T lymphocytes in caprine arthritis-encephalitis lentivirus infection. AB - Peripheral blood mononuclear cells (PBMC) from Saanen goats experimentally infected with the lentivirus caprine arthritis-encephalitis virus (CAEV) were evaluated by semiquantitative reverse transcriptase PCR for gamma interferon (IFN gamma), interleukin-4 (IL-4), and IL-2 gene expression following in vitro stimulation with purified CAEV gp135 surface protein (SU). Studies examined three goats with chronic arthritis and four clinically asymptomatic goats at 5 years postinfection. SU-responsive IFN-gamma mRNA-positive cells and IL-4 mRNA-positive cells in PBMC from infected goats reflected differences in lymphokine balance associated with disease status. IFN-gamma mRNA-positive cells were dominant in PBMC from asymptomatic goats, whereas SU-responsive IL-4 mRNA-positive cells were dominant in PBMC from goats with arthritis. IL-2 gene expression was not responsive to SU stimulation of PBMC from either asymptomatic or arthritic goats. Lymphokine mRNA profiles in SU-stimulated PBMC were dependent on the presence of CD4+ T lymphocytes. The results indicate that asymptomatic goats have a dominant population of CAEV SU-reactive T-helper 1 (Th1)-like lymphocytes in PBMC whereas goats with clinical arthritis have a dominant population of SU-reactive Th2-like lymphocytes. PMID- 9223531 TI - Attenuation of a human rotavirus vaccine candidate did not correlate with mutations in the NSP4 protein gene. AB - The NSP4 protein of a simian rotavirus was reported to induce diarrhea following inoculation of mice. If NSP4 is responsible for rotavirus diarrhea in humans, attenuation of a human rotavirus may be reflected in concomitant mutations in the NSP4 gene. After 33 passages in cultured monkey kidney cells, a virulent human rotavirus (strain 89-12) was found to be attenuated in adults, children, and infants. Nucleotide sequence analysis of the NSP4 protein gene revealed only one base pair change between the virulent (unpassaged) and attenuated 89-12 viruses, which resulted from a substitution of alanine for threonine at amino acid 45 of the encoded NSP4 protein. Because both threonine and alanine have been found at position 45 of NSP4 in symptomatic and asymptomatic human rotaviruses, neither amino acid in this position could be established as a marker of virulence. Therefore, attenuation of rotavirus strain 89-12 appears to be unrelated to mutations in the NSP4 gene. PMID- 9223532 TI - High viral load in semen of human immunodeficiency virus type 1-infected men at all stages of disease and its reduction by therapy with protease and nonnucleoside reverse transcriptase inhibitors. AB - Seminal viral load is likely to be directly related to the sexual transmissibility of human immunodeficiency virus type 1 (HIV-1). However, it is not clear whether the level of HIV-1 in semen varies with the stage of infection and whether antiretroviral therapy reduces seminal viral load. A nucleic acid sequence-based amplification (NASBA) technique was used to quantify HIV-1 RNA as an indicator of infectious viral load in semen and blood plasma of homosexual men with different stages and durations of HIV-1 infection. The median viral load in a cross section of 34 men was 11,000 HIV-1 RNA copies/ml (range, <400 to 1.3 x 10(7) copies/ml) in whole semen and 5,238 HIV-1 RNA copies/ml (range, <400 to 2.8 x 10(5) copies/ml) in seminal plasma, which is 10- to 1,000-fold higher than previous estimates. Viral loads in whole semen and seminal plasma were strongly correlated with blood plasma viral load (P < 0.001) but not with blood CD4+ T cell count (P = 0.420). Longitudinal analysis of eight subjects who progressed to AIDS showed that seminal viral load increased in most cases, with viral load consistently higher in blood plasma than in semen. Viral loads in semen and blood plasma decreased markedly in six other patients following initiation of potent combination therapy with a protease inhibitor (indinavir) and a nonnucleoside reverse transcriptase inhibitor (DMP-266). These findings have important implications for the biology of sexual transmission of HIV-1 and its potential reduction by antiretroviral therapy. PMID- 9223533 TI - The kappa opioid agonist niravoline decreases brain edema in the mouse middle cerebral artery occlusion model of stroke. AB - The effect of niravoline (RU 51599), a kappa opioid receptor agonist with water diuretic properties, was assessed on the resorption of postischemic cerebral edema in the conscious mouse in comparison with U 50488, another kappa opioid receptor agonist, and mannitol. Ischemia was obtained by permanent occlusion of the right middle cerebral artery. Twenty-four hours after occlusion, at a time when brain water content is submaximal, blood samples were collected to measure serum osmolality, and brains were removed to measure the brain water content of two samples of frontoparietal cortical tissue corresponding to the core and the periphery of ischemia. When administered from 3 to 30 mg/kg as a single i.p. injection 20 h after occlusion, niravoline significantly reduced the brain cortical water increase by 27% up to 48% in the periphery of the ischemic tissue. At these same doses, it increased the serum osmolality to the same extent in ischemic as in nonischemic mice: 4 to 10 mOsm/kg. U 50488 generally showed a similar activity. In contrast, mannitol (1 or 2 g/kg i.p. 23 h after occlusion) increased serum osmolality but did not decrease brain water content. In conclusion, kappa opiate agonists could be an alternative to hyperosmotic agents in the treatment of cerebral edema of the focal ischemia type, the use of which is limited to the early phase of cerebral edema. PMID- 9223534 TI - Effect of smoking history on [3H]nicotine binding in human postmortem brain. AB - Chronic nicotine administration in animal models evokes a dose-dependent increase in brain nicotinic receptor numbers. Genetically determined variability in nicotinic receptor number in different mouse strains has also been reported, which is thought to affect sensitivity to nicotine, as well as the development of tolerance. Humans self-administer nicotine principally in the form of cigarettes and other tobacco products. The present study compared [3H]nicotine binding in human postmortem brain from thalamus and hippocampus of nonsmoking subjects, subjects who had variable life-long smoking histories and subjects who had quit smoking. A significant increase was seen in [3H]nicotine binding in both hippocampus and thalamus of subjects with life-long smoking histories. In the hippocampus, this change resulted from a change in total receptor number (Bmax), with no change in receptor affinity (Kd). There was also a positive correlation between the degree of smoking, as measured by the average reported packs smoked per day, and the number of nicotine binding sites found in both the hippocampus and thalamus, showing that humans exhibit a dose-dependent increase in brain nicotinic receptor binding. Receptor levels in these brain regions after smoking cessation were at or below those found in the control population, which indicated that smoking-induced changes are reversible after cessation of nicotine treatment. These results suggest that increases in nicotinic receptor levels in the human brain may underlie nicotine tolerance and addiction in smokers. PMID- 9223535 TI - Opposing effects of vasoactive intestinal polypeptide on gastric motor function in the dorsal vagal complex and the nucleus raphe obscurus of the rat. AB - Vasoactive intestinal polypeptide (VIP)-like immunoreactive cell bodies and fibers and VIP binding sites exist in the brainstem nuclei that regulate autonomic function. Therefore, we investigated the effects of microinjection of VIP in the dorsal vagal complex (DVC), nucleus raphe obscurus (nROb) and nucleus ambiguus of alpha-chloralose-anesthetized rats while recording intragastric pressure, pyloric and greater curvature smooth muscle contractile activity, blood pressure and heart rate. Microinjection of VIP into the DVC increased intragastric pressure (1-100 pmol) and pyloric smooth muscle contractile activity (100 pmol), as well as arterial blood pressure (1-100 pmol). Whereas VIP microinjected into the nROb (10-100 pmol) decreased intragastric pressure and inhibited pyloric smooth muscle contractile activity. Mean arterial blood pressure increased in response to VIP in the nROb at the highest dose of 100 pmol only. No changes in gastric motor and cardiovascular function were noted after microinjection of VIP (1-100 pmol) into the region of the nucleus ambiguus. The gastric motor effects of VIP in the DVC (10 pmol) and nROb (50 pmol) were completely abolished by bilateral cervical vagotomy. These data show that VIP may produce opposite vagally mediated gastric motor responses upon its microinjection into the DVC and nROb. PMID- 9223536 TI - The cardiac effects of pimobendan (but not amrinone) are preserved at rest and during exercise in conscious dogs with pacing-induced heart failure. AB - We compared the effects of pimobendan (0.25 mg/kg i.v.), a Ca++ sensitizer, with some phosphodiesterase-III inhibition effects, and amrinone (1 mg/kg plus 10 microg/kg/min i.v.), a PDE-III inhibitor, on left ventricular (LV) systolic and diastolic performance, both at rest and during exercise, in seven conscious dogs before and after pacing-induced congestive heart failure (CHF). Before CHF, under resting conditions, both pimobendan and amrinone caused a similar significant decrease in left ventricle size and end-systolic pressure, arterial elastance, and the time constant of LV relaxation. Similar results were obtained during exercise. Both agents also produced a similar increase in E(ES), the slope of the LV end-systolic pressure-volume relation (3.4 +/- 1.5 vs. 4.2 +/- 1.1 mm Hg/ml; amrinone vs. pimobendan). After CHF, the vasodilatory effects of amrinone and pimobendan were preserved both at rest and during exercise; however, the inotropic actions were different. After CHF, pimobendan increased E(ES) (3.9 +/- 0.5 vs. 5.7 +/- 0.4 mm Hg/ml, P < .05), decreased the time constant of LV relaxation, increased the maximum rate of LV filling (37 +/- 19 ml/sec) (P < .05) and produced a downward shift of the early diastolic portion of LV pressure volume loop. Pimobendan also augmented LV contractile performance during CHF exercise. In contrast, after CHF, amrinone no longer produced a positive inotropic effect. Amrinone improved LV relaxation and filling, both at rest and during exercise after CHF, but significantly less than pimobendan. We conclude that after CHF, the cardiac response to a PDE-III inhibitor is attenuated, but the response to Ca++ sensitizer is preserved. Thus, after CHF, pimobendan is more effective than amrinone in enhancing LV contractile state, LV relaxation and LV filling both at rest and during exercise. PMID- 9223537 TI - Pharmacological comparison of transient and persistent [3H]dopamine release from mouse striatal synaptosomes and response to chronic L-nicotine treatment. AB - L-Nicotine stimulates a biphasic release of [3H]dopamine from mouse striatal synaptosomes which does not persist after agonist is removed. Approximately 80% of the initial release is transient and disappears with a half-time of less than 1 min; the other 20% persists for several minutes (t(1/2), 5-10 min). Both the transient and persistent phases were investigated by 10-min exposures to agonists with an in vitro perfusion technique. A series of nicotinic agonists and antagonists were used to determine the pharmacological relationship of the two phases. Parameters measured included EC50 and Vmax values and desensitization rates for both phases for agonists, Ki values for antagonists and Ki values for low concentrations of agonists. The results are consistent with both phases being mediated by a single type of receptor. In addition, the effects of chronic nicotine treatment on transient and persistent [3H]DA release were measured. For both phases, release was decreased approximately 15% by chronic infusion of 4.0 mg/kg/hr L-nicotine. Correlation of the results with inactivation of a portion of the receptors rather than a reversible desensitization is discussed. PMID- 9223538 TI - Effects of kappa opioids on cocaine self-administration by rhesus monkeys. AB - Kappa opioid agonists attenuate some neurochemical and behavioral effects of cocaine and are being considered as potential treatments for cocaine dependence. The present study examined the effects of two kappa opioid agonists, the benzomorphan ethylketocyclazocine (EKC) and the arylacetamide U50,488, on cocaine self-administration in rhesus monkeys. Monkeys responded for 0.032 mg/kg/injection cocaine (i.v.) and 1 g banana-flavored food pellets during alternating daily sessions of cocaine and food availability. Chronic treatment for 10 consecutive days with EKC (0.0032-0.032 mg/kg/hr) or U50,488 (0.032-0.1 mg/kg/hr) dose-dependently decreased self-administration of cocaine unit doses at the peak of the cocaine dose-effect curve (0.01 and 0.032 mg/kg/injection). These decreases in cocaine self-administration were often sustained throughout the 10 days of treatment. Doses of EKC and U50,488 that decreased cocaine self administration usually decreased food-maintained responding as well. In addition, EKC and U50,488 often produced emesis and sedation during the first few days of treatment, although tolerance appeared to develop rapidly to these effects. In general, EKC produced fewer undesirable effects than U50,488 at doses that decreased cocaine self-administration. The kappa antagonist norbinaltorphimine (3.2 mg/kg) did not affect responding maintained by cocaine or food. However, both norbinaltorphimine (3.2 mg/kg) and the opioid antagonist naloxone (1.0 mg/kg/hr) blocked the effects of EKC and U50,488. These results indicate that chronic administration of EKC and U50,588 produce a dose-dependent, kappa receptor-mediated and often sustained decrease in cocaine self-administration. However, these kappa agonists also produce undesirable behavioral effects that may complicate their use as treatments for cocaine dependence. PMID- 9223539 TI - In vitro and in vivo characterization of the dopamine D4 receptor, serotonin 5 HT2A receptor and alpha-1 adrenoceptor antagonist (R)-(+)-2-amino-4-(4 fluorophenyl)-5-[1-[4-(4-fluorophenyl)-4-oxobutyl] pyrrolidin-3-yl]thiazole (NRA0045). AB - (R)-(+)-2-Amino-4-(4-fluorophenyl)-5-[1-[4-(4-fluorophenyl)-4-oxobutyl]+ ++pyrrolidin-3-yl]thiazole (NRA0045), a novel thiazole derivative, has high affinities for the human cloned dopamine D4.2, D4.4 and D4.7 receptors, with Ki values of 2.54, 0.55 and 0.54 nM, respectively. NRA0045 is approximately 91-fold more potent at the dopamine D4.2 receptor, compared with human cloned dopamine D2L receptor. NRA0045 also has high affinities for the serotonin (5-HT)2A receptor (Ki = 1.92 nM) and alpha-1 adrenoceptor (Ki = 1.40 nM) but weak affinities (IC50 values are approximately 1 microM) for six other neurotransmitter receptors (adenosine1, 5-HT1A, 5-HT1C, dopamine transporter, alpha2A and alpha2A) and negligible affinities (IC50 values are over 10(-5) M) for 42 other receptors, including neurotransmitters and hormones, ion channels and second messenger systems. Locomotor hyperactivity induced by methamphetamine (1 mg/kg i.p.) in mice was dose-dependently antagonized by NRA0045 (ED50 = 0.5 mg/kg i.p. and 1.9 mg/kg p.o., respectively). Methamphetamine (10 mg/kg i.p.) induced stereotyped behavior in mice was dose-dependently antagonized by NRA0045, whereas NRA0045 did not exceed 50% inhibition even at the highest dose given (30 mg/kg i.p.). Catalepsy was dose-dependently and significantly induced by NRA0045 in rats, whereas NRA0045 did not exceed 50% induction even at the highest dose given (30 mg/kg i.p.). Thus NRA0045 blocks behaviors associated with activation of the mesolimbic/mesocortical dopaminergic neurons more selectively than behaviors associated with nigrostriatal dopaminergic neurons. In rats, tryptamine induced clonic seizure, a 5-HT2 receptor-mediated behavior, was also dose dependently inhibited by NRA0045 (ED50 = 1.7 mg/kg i.p.). Norepinephrine-induced lethality is regarded as being induced through the alpha-1 adrenoceptor. NRA0045 dose-dependently antagonized norepinephrine-induced lethality in rats (ED50 = 0.2 mg/kg i.p.). Thus NRA0045 may have a unique antipsychotic activity with regard to dopamine D4 and 5-HT2A receptors and alpha-1 adrenoceptor antagonistic activities, without producing the extrapyramidal side effects. PMID- 9223540 TI - Action of methylmercury on GABA(A) receptor-mediated inhibitory synaptic transmission is primarily responsible for its early stimulatory effects on hippocampal CA1 excitatory synaptic transmission. AB - Bath application of methylmercury (MeHg) causes an early stimulation before block of synaptic transmission in the CA1 region of hippocampal slices. Effects of MeHg and Hg++ on inhibitory postsynaptic potentials (IPSPs) or currents (IPSCs) and excitatory postsynaptic potentials (EPSPs) or currents (EPSCs) were compared to test whether or not early block by MeHg of GABA(A)-mediated inhibitory synaptic transmission and MeHg-induced alterations of the resting membrane potentials of CA1 neurons contribute to this initial enhancement of excitability. MeHg affected IPSPs and IPSCs similarly, and more rapidly than EPSPs and EPSCs. In contrast, although Hg++ blocked IPSPs more rapidly than EPSPs, times to block of IPSCs and EPSCs by Hg++ were virtually identical when CA1 neurons were voltage-clamped at their resting membrane potential levels. MeHg increased EPSC amplitudes before their subsequent decrease even when CA1 neuronal membranes were voltage-clamped at their resting potentials. This suggests that effects of MeHg on CA1 cell membrane potentials are not a major factor for MeHg-induced early stimulation of hippocampal synaptic transmission. Effects of MeHg and Hg++ on the reversal potentials for IPSCs also differed. Both metals blocked all outward and inward currents generated at different holding potentials. However, MeHg shifted the current-voltage (I/V) relationship to more positive potentials, although Hg++ shifted the I/V curve to more negative potentials. Hg++ was a less potent blocker of on IPSCs and EPSPs or EPSCs than was MeHg. To determine if the early increase in amplitude of population spikes or EPSPs is due to an action of MeHg at GABA(A) receptors, extracellular recordings of population spikes and intracellular recordings of EPSPs were compared with or without pretreatment of hippocampal slices with bicuculline. After preincubation of slices with 10 microM bicuculline for 30 to 60 min, MeHg only decreased the amplitudes of population spikes and EPSPs to block; no early increase of synaptic transmission occurred. Pretreatment of slices with strychnine, did not prevent MeHg-induced early increase in population spikes. MeHg also blocked responses evoked by bath application of muscimol, a GABA(A) agonist. Thus, block by MeHg of GABA(A) receptor-mediated inhibitory synaptic transmission may result in disinhibition of excitatory hippocampal synaptic transmission, and appears to be primarily responsible for the initial excitatory effect of MeHg on hippocampal synaptic transmission. PMID- 9223541 TI - Waglerin-1 modulates gamma-aminobutyric acid activated current of murine hypothalamic neurons. AB - We examined the effect of Waglerin-1, a peptide of 22 amino acid residues purified from the venom of Wagler's pit viper (Trimeresurus wagleri), on the whole cell current response (I(GABA)) of freshly isolated murine hypothalamic neurons to gamma-aminobutyric acid (GABA). Although the application of 32 microM Waglerin-1 alone had no effect on membrane conductance, coapplication with GABA increased I(GABA) for 78 and suppressed I(GABA) for 44 of the 141 neurons examined. The potentiating effect of Waglerin-1 was associated with a leftward shift of the concentration-response relation of GABA without increasing peak I(GABA). This potentiating effect of Waglerin-1 on I(GABA) mimics diazepam. Furthermore, the benzodiazepine antagonist flumazenil antagonized Waglerin-1 potentiation of I(GABA), These observations suggest that Waglerin-1 acts on the benzodiazepine site of one type of GABA(A) receptor/channel complex to increase its affinity for agonist. In contrast, the depressant effect of Waglerin-1 was associated with a rightward shift of the concentration-response relation of GABA without depressing the maximal I(GABA); this suggests a competitive inhibition of a second class of GABAR. The ability of Waglerin-1 to suppress I(GABA) showed a positive correlation with a similar action of Zn++. As with Zn++, the depressant effect of Waglerin-1 on I(GABA) was more pronounced at negative holding potentials. These observations are discussed in terms of variation in the subunit composition of GABA receptors that murine central nervous system neurons express. PMID- 9223542 TI - Transport of clonidine across cultured brain microvessel endothelial cells. AB - Brain penetration of clonidine, an alpha-2 adrenoceptor agonist, was studied using an in vitro cell culture system consisting of primary cultures of porcine brain capillary endothelial cells. Uptake of clonidine was measured as a function of its concentration in the incubation mixture. Saturation of uptake was apparent and could be described by Michaelis-Menten-type kinetics (K(M) = 1.34 mM; Vmax = 0.099 nmol/min/cm2). Saturation was not observed at a low temperature (4 degrees C). Transendothelial transport experiments revealed that translocation of clonidine cannot be attributed solely to paracellular leakage. Uptake was reduced at low extracellular pH or by using an incubation buffer that contained the K+ ionophore valinomycin. Time-dependent uptake of clonidine and transendothelial transport were slower than expected considering the high octanol-to-buffer partition coefficient of this compound. On the basis of transendothelial transport experiments, we concluded that the carrier system responsible for active transport of clonidine is located at both the apical and the basolateral membrane domain. PMID- 9223543 TI - Enhancement of analgesia from systemic opioid in humans by spinal cholinesterase inhibition. AB - Intravenous opioids cause analgesia and increase release of ACh in spinal cord dorsal horn in animals, and these effects are enhanced by intrathecal neostigmine injection. The purpose of the current study was to test whether intrathecal neostigmine enhanced analgesia and increased cerebrospinal fluid concentrations of ACh over those induced by i.v. alfentanil in volunteers, and also to test whether neostigmine enhanced alfentanil-induced side effects. After human studies committee approval, 40 healthy volunteers received an intrathecal injection of saline or neostigmine (50, 100 or 200 microg) followed in 60 min by a computer controlled, stepped i.v. infusion of alfentanil to escalating targeted plasma concentrations. Pain report to hand and foot immersion in ice water, sedation, nausea, weakness, vital signs, end-tidal CO2 and oxyhemoglobin saturation were measured 60 min after spinal injection and at the end of each 20-min alfentanil infusion. Cerebrospinal fluid was sampled once after drug administration. Intrathecal neostigmine alone caused analgesia in the foot but not in the hand, and was accompanied by leg weakness, whereas IV alfentanil alone caused equivalent analgesia in both the hand and the foot and was accompanied by nausea, sedation, increased end-tidal CO2 and decreased oxyhemoglobin saturation. Neostigmine enhanced analgesia but not respiratory effects induced by i.v. alfentanil; it also enhanced nausea and sedation. Intravenous alfentanil increased cerebrospinal fluid ACh concentration, and neostigmine enhanced this change. These data in humans are consistent with a spinal cholinergic mechanism of i.v. opioid analgesia. Because neostigmine enhances both analgesia and side effects induced by i.v. alfentanil, the clinical utility of their use in combination will depend on the relative strength of these interactions. PMID- 9223544 TI - Evaluation of the antimyotonic activity of mexiletine and some new analogs on sodium currents of single muscle fibers and on the abnormal excitability of the myotonic ADR mouse. AB - To search for use-dependent sodium channel blockers to selectively solve skeletal muscle hyperexcitability in hereditary myotonias, mexiletine (MEX; compound I) and its newly synthetized analogs, 2-(4-chloro-2-methylphenoxy)-benzenethanamine (compound II) and (-)-S-3-(2,6-dimethylphenoxy)-2-methylpropanamine (compound III), were tested on intercostal muscle fibers from the myotonic ADR mouse through use of the standard current-clamp microelectrode technique. In parallel, the effects of these compounds on the sodium channels were measured on frog muscle fibers under voltage-clamp conditions. The tonic and use-dependent blocks of peak sodium currents (I(Namax)) produced by each compound were evaluated by using a single depolarizing pulse and a pulse train at 10 Hz frequency, respectively. At 10 and 50 microM, MEX decreased the occurrence of spontaneous excitability in myotonic muscle fibers; 100 microM was required to decrease the amplitude of the action potential and the stimulus-induced firing of the membrane as well as to increase the threshold for generation of action potential. At 300 microM, MEX decreased the latency of the action potential and increased the threshold current to elicit a single action potential. MEX produced a tonic block of I(Namax) with an half-maximal concentration (IC50) of 83 microM, but the IC50 value for use-dependent block was 3-fold lower. Compound III, which differs from MEX in that it has a longer alkyl chain, similarly blocked first the spontaneous and then the stimulus-evoked excitability of myotonic muscle fibers but at 2-fold lower concentrations than MEX. Compound III was less potent than MEX in producing a tonic block of I(Namax) (IC50 = 108 microM) but was a strong use-dependent blocker with an IC50 close to 15 microM. The more lipophylic compound II irreversibly blocked both spontaneous and stimulus-evoked membrane excitability at concentrations as low as 10 microM and shortened the latency of the action potential in a concentration-dependent fashion. Compound II produced a potent tonic block of I(Namax) (IC50 = 30 microM), and its potency increased 2-fold during high-frequency stimulation. Both of the new analogs (compound II in particular), but not MEX, were less effective on the excitability parameters of striated fibers of healthy vs. ADR mice, a characteristic that increases their interest as potential antimyotonic agents. PMID- 9223546 TI - Heme polymerase activity and the stage specificity of antimalarial action of chloroquine. AB - Plasmodium falciparum lysate, prepared from 2.7 x 10(7) ring-infected erythrocytes and incubated with hemoglobin in sodium acetate at pH 5, incorporated a mean of 1.6 nmol of ferriprotoporphyrin IX (FP) into hemozoin in 18 to 22 hr. A similar preparation of trophozoite lysate incorporated a mean of 3.6 nmol of FP into hemozoin in 4 to 6 hr. These findings indicate differences between heme polymerase activity (hemozoin production) at the ring and trophozoite stages of malaria parasites. Intracellular hemozoin production was 90% inhibited at the ring and trophozoite stages by 0.5 and 7 nmol of chloroquine/10(6) infected erythrocytes. respectively. The inhibition killed the rings but not the trophozoites, suggesting that mature parasites may have a mechanism for protecting themselves against chloroquine-FP toxicity. PMID- 9223545 TI - Analysis of eicosanoid mediation of the renal functional effects of hyperchloremia. AB - Depression of GFR and antinatriuresis in response to high chloride has been linked to a cyclooxygenase (COX)-dependent mechanism involving thromboxane A2 (TxA2) and prostaglandin endoperoxide (PGH2), because inhibition of COX prevented the fall in GFR and antinatriuresis produced by hyperchloremia. However, hyperchloremia did not increase, but unexpectedly decreased, renal prostaglandin and TxA2 efflux (Yin et al., 1995). To resolve questions regarding the role of eicosanoids in mediating the renal functional effects of high chloride (117 mM), by stimulating either TxA2 synthesis or TxA2/PGH2 receptors, we compared the ability of indomethacin to block high-chloride effects in the rat isolated kidney with that of BMS 180291 and SQ 29548, antagonists of the TxA2/PGH2 receptor. These antagonists differ in terms of their selectivity and their capacity to inhibit isoforms of the TxA2/PGH2 receptor. Indomethacin and SQ 29548 had identical actions, preventing the decrease of GFR and antinatriuresis evoked by hyperchloremia, e.g., sodium excretion rate in the SQ 29548 and indomethacin groups increased to 7.2 +/- 1.3 and 7.1 +/- 1.2 microEq/min, respectively, compared with 2.6 +/- 0.7 microEq/min in the control group. In contrast, neither BMS 180291 nor the TxA2 synthase inhibitors, OKY 046 and CGS 13080, modified the negative effects of high chloride on GFR or sodium excretion. These results argue against either TxA2 or PGH2 acting as mediator of the effects of high chloride on renal function and suggest a product of COX activity such as a 20-HETE analog of prostaglandin endoperoxide. Evidence to support this proposal was obtained: 1) Hyperchloremia increased 20-HETE release from the rat kidney by 2-fold when compared with low-chloride conditions of renal perfusion. 2) The renal vasoconstrictor action of 20-HETE was shown to be dependent on COX activity and to be antagonized by blockade of the TxA2/PGH2 receptor. PMID- 9223547 TI - Signal transduction mechanism(s) involved in prostacyclin production elicited by acetylcholine in coronary endothelial cells of rabbit heart. AB - The purpose of this study was to elucidate the mechanism by which acetylcholine (ACh) promotes prostacyclin (PGI2) production in cultured coronary endothelial cells (CEC) of the rabbit heart. ACh-induced production of PGI2, measured as immunoreactive 6-keto-PGF1alpha, was enhanced by increasing the extracellular calcium (Ca++) concentration and reduced by Ca++ depletion. The receptor-operated Ca++ channel blocker SK&F96365, but not the voltage-dependent Ca++ channel blockers verapamil or nifedipine, attenuated ACh-induced 6-keto-PGF1alpha production and the associated rise in cytosolic Ca++. Thapsigargin, which depleted Ca++ accumulation from the intracellular Ca++ store, did not prevent the ACh-induced rise in cytosolic Ca++. In the absence of extracellular Ca++, ACh and ATP increased cytosolic Ca++ but did not alter 6-keto-PGF1alpha production. In permeabilized CEC, guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S) but not ACh enhanced 6-keto-PGF1alpha synthesis. ACh increased 6-keto-PGF1alpha production in the presence of GTP-gamma-S. These effects of GTP-gamma-S were attenuated by guanosine 5'-O-(2-thiotriphosphate). In the absence of extracellular Ca++, ACh or ATP increased cytosolic Ca++ in cells permeabilized with beta-escin and loaded with GTP-gamma-S; this effect was attenuated by guanosine 5'-O-(2 thiotriphosphate). The effect of ATP but not ACh to mobilize intracellular Ca++ or increase 6-keto-PGF1alpha was inhibited by pertussis toxin. The phospholipase C inhibitor D609, which attenuated ACh- and ATP-induced mobilization of intracellular Ca++, did not alter 6-keto-PGF1alpha production. The NO synthase inhibitor N-monomethyl-arginine also failed to alter ACh-induced 6-keto-PGF1alpha synthesis. These data suggest that, in CEC of the rabbit heart, ACh stimulates prostacyclin production via a pertussis toxin-insensitive G protein and by increasing the influx of extracellular Ca++ through a G protein-independent receptor-operated Ca++ channel. PMID- 9223548 TI - Variabilin: a dual inhibitor of human secretory and cytosolic phospholipase A2 with anti-inflammatory activity. AB - The marine product variabilin was identified as a novel inhibitor of phospholipase A2 (PLA2), which exhibited IC50 values of 6.9 microM and 7.9 microM for human synovial secretory PLA2 and U937 cells cytosolic PLA2 activities, respectively. This compound was less potent on bee venom or zymosan-injected rat air pouch enzymes and failed to affect Naja naja venom PLA2. The production of leukotriene B4 by human neutrophils stimulated with calcium ionophore A23187 was also inhibited by variabilin, which was without effect on 5-lipoxygenase, cyclo oxygenase 1 and cyclo-oxygenase 2 activities in cell-free assays. Other functions of human neutrophils, such as degranulation and superoxide generation, were also significantly reduced in vitro. Variabilin administered topically suppressed the mouse ear edema induced by 12-O-tetradecanoylphorbol 13-acetate, whereas the ear edema induced by arachidonic acid was unaffected; this suggests an action previous to arachidonic acid metabolism. This compound administered p.o. at 30 mg/kg and 45 mg/kg significantly inhibited mouse paw edema induced by carrageenan and, at 0.01 to 1.0 micromol/pouch in the mouse air pouch injected with zymosan, exerted a marked inhibition on PGE2 and leukotriene B4 levels in exudates (ID50 values of approximately 0.028-0.029 micromol/pouch), without affecting cell migration. Our results indicate that variabilin is an inhibitor of human secretory and cytosolic PLA2 activities that controls eicosanoid production in vitro and in vivo, inhibits neutrophil degranulation and superoxide generation in vitro and shows anti-inflammatory activity after topical or p.o. administration to mice. PMID- 9223549 TI - S 15535, a novel benzodioxopiperazine ligand of serotonin (5-HT)1A receptors: I. Interaction with cloned human (h)5-HT1A, dopamine hD2/hD3 and h alpha2A adrenergic receptors in relation to modulation of cortical monoamine release and activity in models of potential antidepressant activity. AB - The novel, potential anxiolytic, S 15535 (4-(benzodioxan-5-yl)1-(indan-2 yl)piperazine), is an agonist and antagonist (weak partial agonist) at pre- and postsynaptic serotonin (5-HT)1A receptors, respectively. Herein, we characterized its influence on dialysate levels of 5-HT, dopamine (DA) and NAD simultaneously determined in single samples of the frontal cortex (FCX) of freely moving rats, and compared its activity in several other models of potential antidepressant (AD) properties with those of the 5-HT reuptake inhibitor (SSRI), fluoxetine. S 15535 displayed high affinity at cloned human (h) 5-HT1A receptors (Ki = 0.7 nM) and >250-fold lower affinity at cloned hD2 (400 nM), hD3 (248 nM) and h alpha2A adrenergic (AR) (190 nM) receptors. S 15535 (0.08-5.0 mg/kg s.c.) markedly and dose-dependently suppressed dialysate levels of 5-HT in the FCX, nucleus accumbens and striatum of freely moving rats, whereas fluoxetine (10.0 mg/kg s.c.) elevated levels of 5-HT in each structure. In contrast to 5-HT, dialysate levels of DA and NAD in the FCX were dose-dependently increased by S 15535, and this effect was mimicked by fluoxetine. The influence of S 15535 and fluoxetine on FCX levels of DA was regionally specific inasmuch as dialysate levels of DA in the accumbens and striatum were not modified. The selective 5-HT1A antagonist, WAY 100,635 (N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl) cyclohexanecarboxamide (0.16) transiently elicited a slight increase in cortical levels of 5-HT, an action opposite to that of S 15535. Further, in the presence of WAY 100,635 (0.16), the influence of S 15535 (0.63) on cortical levels of 5 HT, DA and NAD was markedly attenuated. Upon chronic administration of S 15535 or fluoxetine (10.0 mg/kg s.c. daily for 14 days, in each case), there was no significant alteration in the density of beta-AR receptors in the FCX. However, in contrast to fluoxetine, S 15535 elicited a significant (25%) decrease in the density (Bmax) of 5-HT2A receptors labeled by [3H]ketanserin in the cortex; there was no alteration in Kd. In a learned helplessness paradigm in rats, S 15535 (0.63-40.0 mg/kg p.o.) markedly reduced escape deficits on each of three consecutive days of testing. Fluoxetine (2.0-8.0 mg/kg i.p.) was also active in each session, but presented a biphasic dose-response curve. Finally, under the conditions used, neither S 15535 (0.63-10.0) nor fluoxetine (0.63-10.0) decreased immobility time in the forced swim test. In conclusion, S 15535 is a selective ligand of cloned, h5-HT1A receptors. Its agonist actions at 5-HT1A autoreceptors underlie its ability to decrease extracellular levels of 5-HT in the FCX, and likely contribute to the increase in extracellular levels of DA and NAD evoked by S 15535 in this structure. Further, S 15535 is active in several other, although not all, models of potential AD activity. Thus, although S 15535 is under development as an anxiolytic agent, a further characterization of its putative AD actions would be of interest. PMID- 9223551 TI - Carrier-mediated hepatic uptake of quinolone antibiotics in the rat. AB - The systemic clearance of many quinolone antibiotics is mainly via metabolism and urinary excretion; by contrast, biliary excretion is a major route of elimination for a new quinolone grepafloxacin (GPFX). Accordingly, we studied the hepatic uptake of GPFX because it is the first step in the drug's hepatobiliary transport. The hepatic uptake of GPFX in vivo after i.v. administration was found to approach the hepatic blood flow, suggesting the existence of an effective hepatic uptake mechanism. To clarify this transport mechanism, GPFX uptake by isolated rat hepatocytes was examined and found to consist of a saturable component (Km 173 microM, Vmax 6.96 nmol/min/mg) and a nonspecific diffusion component. The inhibition of GPFX uptake by ATP-depletors and a lack of effect after replacing Na+ with choline demonstrated that the uptake was an Na+ independent carrier-mediated active process. This uptake was inhibited by other quinolones and for lomefloxacin this was competitive in nature. Mutual inhibition studies were undertaken to investigate whether the transporter for GPFX might be the same as other transporters so far identified. GPFX inhibited the uptake of taurocholic acid, pravastatin (organic anion), cimetidine (organic cation) and ouabain (neutral steroid). However, GPFX uptake was not inhibited by these compounds. Confirmation that GPFX uptake is blood flow limited was obtained by extrapolation of the in vitro data based on mathematical modeling. In conclusion, the effective hepatic uptake of quinolone antibiotics are via carrier-mediated active transport, which is distinct from that involved in the transport of bile acids, organic anions, organic cations or neutral steroids. PMID- 9223550 TI - S 15535, a novel benzodioxopiperazine ligand of serotonin (5-HT)1A receptors: II. Modulation of hippocampal serotonin release in relation to potential anxiolytic properties. AB - In these studies, we characterized the influence of the novel benzodioxopiperazine serotonin (5-HT)1A ligand, S 15535, on the release of 5-HT in rat hippocampus and compared its potential anxiolytic properties with those of the 5-HT1A receptor partial agonist, buspirone, the 5-HT1A antagonist, WAY 100,635 and the benzodiazepine, diazepam (DZM). (Doses are in milligrams per kilogram s.c., unless otherwise specified.) S 15535 dose-dependently (0.3-3.0) reduced dialysate concentrations of 5-HT in the hippocampus of anesthetized rats. This action of S 15535 (3.0) was blocked by WAY 100,635 (0.3), (-)-penbutolol (2.0) and (-)-tertatolol (8.0), antagonists at 5-HT1A autoreceptors. In rats, fear-induced ultrasonic vocalizations (USVs) were dose-dependently abolished by S 15535 (0.16-2.5 s.c. and 0.63-10.0 p.o.), an action mimicked by buspirone (0.02 2.5) and DZM (0.16-10.0). Further, the action of S 15535 (0.63) was abolished by WAY 100,635 (0.16) and (-)-penbutolol (10.0), which were inactive alone. S 15535 dose-dependently (0.63-10.0 s.c. and 2.5-40.0 p.o.) blocked aggressive encounters in isolated mice; buspirone (0.16-10.0) and, at high doses, DZM (2.5-40.0) were also effective. WAY 100,635 (0.16), which was inactive alone, fully antagonized the antiaggressive actions of S 15535 (2.5). In an elevated plus-maze, neither S 15535 (0.0025-10.0), buspirone (0.0025-10.0) nor WAY 100,635 (0.00063-0.63) significantly increased open-arm entries, whereas they were increased by DZM (0.16-0.63). In the pigeon conflict test, S 15535 (0.04-0.16 i.m.) markedly increased punished responses and only slightly decreased unpunished responses, even at a 64-fold higher dose. In contrast, buspirone (0.16-2.5 i.m.) and DZM (0.04-2.5 i.m.) showed no or a less marked (4-fold) separation between doses increasing punished and decreasing unpunished responses. In the presence of the 5 HT1A antagonist, (-)-alprenolol (10.0 mg/kg i.m.), S 15535 did not increase punished responses. In a Geller conflict paradigm in rats, S 15535 dose dependently (0.3-3.0) increased punished responses, and its action (1.0) was blocked by (-)-penbutolol (8.0). S 15535 (0.63-40.0 s.c. and 2.5-40.0 p.o.) exerted little influence on motor behavior. In conclusion, in line with its net inhibition of serotoninergic transmission by activation of 5-HT1A autoreceptors and blockade of postsynaptic 5-HT1A receptors, S 15535 expresses anxiolytic activity. In addition, it displays antiaggressive (and antidepressant, accompanying paper) properties. Further, S 15535 does not compromise motor behavior at doses over which it expresses its anxiolytic properties. Thus, S 15535 represents a promising candidate for the treatment of anxious states in man. PMID- 9223552 TI - Full reversal of Pb++ block of L-type Ca++ channels requires treatment with heavy metal antidotes. AB - The mechanisms of Pb++ block and unblock of L-type Ca++ channel currents were measured using ventricular myocytes or the cloned channel. The cloned channel was expressed in either Xenopus laevis oocytes or human embryonic kidney cells (HEK 293, stable transfectants). The threshold for Pb++ block was 1 nM, and the apparent IC50 value was 152 nM in oocytes and 169 nM in HEK 293 cells. Pb++ block was dependent on the composition of the external recording solution but not dependent on the subunit composition of the channel. Pb++ block was voltage dependent, with little block observed at negative test potentials using low concentrations of Pb++. Strong depolarizations (>+100 mV) reversed Pb++ block, allowing measurement of reblock kinetics. Reblock was fast (tau = 11 msec), as measured during a +20-mV test pulse. Simple washout did not completely reverse Pb++ block, especially after exposure to concentrations of >100 nM. Full recovery could only be observed after treatment with heavy metal antidotes such as meso 2,3-dimercaptosuccinic acid, 2,3-dimercapto-1-propanesulfonic acid and EDTA. These results suggest that Pb++ blocks voltage-gated Ca++ channels by two mechanisms and that full reversal of lead block requires chelator treatment. PMID- 9223553 TI - [35S]Guanosine-5'-O-(3-thio)triphosphate binding as a measure of efficacy at human recombinant dopamine D4.4 receptors: actions of antiparkinsonian and antipsychotic agents. AB - Recombinant human dopamine D4.4 receptor-mediated G protein activation was characterized in membranes of transfected mammalian (Chinese hamster ovary) cells by the use of [35S]guanosine-5'-O-(3-thio)triphosphate ([35S]GTPgammaS) binding. An initial series of experiments defined the conditions (3 microM GDP, 100 mM NaCl, 3 mM MgCl2) under which optimal stimulation (2.2-fold increase in specific [35S]GTPgammaS binding) was achieved with the endogenous agonist dopamine. The number of dopamine-activated G proteins in Chinese hamster ovary-D4.4 membranes was determined through [35S]GTPgammaS isotopic dilution saturation binding, yielding a Bmax value of 2.29 pmol/mg. This compared with a D4.4 receptor Bmax value of 1.40 pmol/mg determined by [3H]spiperone saturation binding, indicating that 1 or 2 G proteins were activated per D4.4 receptor and that there were few or no "spare receptors" in this cell line. Under these conditions, the efficacy for stimulation of [35S]GTPgammaS binding at D4.4 receptors of 12 dopaminergic agonists was determined. Several antiparkinsonian drugs, including ropinirole, quinerolane and lisuride, exhibited agonist activity at D4.4 receptors (Emax = 74.3%, 72.4% and 32.2%, respectively, compared with dopamine = 100%). The EC50 values for agonist stimulation of [35S]GTPgammaS binding correlated well with the inhibition constants derived from competition binding with [3H]spiperone (r = +.99). However, other antiparkinsonian drugs (bromocriptine, L-DOPA and terguride) showed low affinity and/or were devoid of agonist activity at D4.4 receptors. The potency at D4.4 receptors of the novel, selective D4.4 receptor antagonist L 745,870 was determined, indicating that it has high affinity (Ki = 1.99 nM) without detectable agonist activity. Furthermore, L 745,870 completely inhibited dopamine-stimulated [35S]GTPgammaS binding with a Kb value of 1.07 nM. The action of an additional 20 chemically diverse dopaminergic ligands, including clozapine, ziprasidone, sertindole, olanzapine and several other "atypical" antipsychotics, in advanced development was investigated. Each of these ligands shifted the dopamine stimulation curve to the right in a parallel manner consistent with competitive antagonism at this site and yielding Kb values (32.6, 22.4, 17.2 and 26.5 nM, respectively) that agreed closely with their Ki values (38.0, 14.9, 18.5 and 26.1 nM). In contrast, raclopride and seroquel exhibited low affinity at D4.4 receptors (Ki > 1000 nM). Other compounds that showed antagonist activity at D4.4 receptors included the 5-hydroxytryptamine2A receptor antagonist fananserin (RP 62203), the sigma ligand BMY 14,802 and the D3 receptor antagonist GR 103,691. In conclusion, dopamine D4.4 receptor activity is unlikely to be an important factor in the clinical effectiveness of antiparkinsonian drugs, although low agonist efficacy at D4.4 receptors might be associated with a lesser incidence of side effects. Furthermore, antagonist activity at D4.4 receptors is a common property of many typical and atypical antipsychotic agents. PMID- 9223554 TI - DMPS-arsenic challenge test. I: Increased urinary excretion of monomethylarsonic acid in humans given dimercaptopropane sulfonate. AB - The purpose of the present study was to evaluate in a novel manner the arsenic exposure of humans living in two towns in Northeastern Chile. Residents of one town drink water containing 593 microg As/l. Those in the control town drink water containing 21 microg As/l. Our hypothesis was that the administration of the chelating agent, 2,3-dimercaptopropane-1-sulfonic acid, Na salt (DMPS, DIMAVAL) would increase the urinary excretion of arsenic, alter the urinary profile of arsenic species and thus result in a better indication of the body load of arsenic and a better biomarker for arsenic exposure. The method used to evaluate these subjects was to give them 300 mg DMPS by mouth, after an overnight fast, and collect urine at specified time periods. The urine samples were analyzed for inorganic arsenic, monomethylarsonic acid (MMA), dimethylarsinic acid (DMA) and total arsenic by hydride generation and atomic absorption spectrophotometry. The results indicated that: 1) During the 2-hr period after DMPS administration, MMA represented 42%, inorganic As, 20 to 22% and DMA, 37 to 38% of the total urinary arsenic. The usual range of the MMA percentage in human urine has been 10 to 20%. The % MMA increased almost equally for both the arsenic exposed and control subjects. 2) The exposed subjects had a greater urinary excretion of total arsenic, before and after DMPS administration, than the control subjects. 3) Although buccal cells were obtained only from a few subjects, the prevalence of mononucleated buccal cells, an indication of genotoxicity, was 5-fold greater for those who consumed drinking water with the higher arsenic content than among control subjects. Our conclusions are that 1) DMPS has a highly specific effect in humans on MMA metabolism and/or urinary excretion; 2) the human body stores substantial amounts of arsenic; and 3) the urinary arsenic concentration after DMPS administration may be more indicative of the body burden of arsenic because it was greater than that found before DMPS was given. PMID- 9223555 TI - An analysis of the mechanisms involved in the okadaic acid-induced contraction of the estrogen-primed rat uterus. AB - The contractile effect of okadaic acid (OA) and its derivatives was investigated in the rat uterus. OA (20 microM) induced a transient contraction which, after plateauing, slowly decreased. The structurally related compound okadanol (20 microM) failed to induce any significant contraction. Conversely, the synthetic compound methyl okadaate (20 microM) and the naturally occurring ester 7'-hydroxy 4'-methyl-2'-methylen-hept-4'(E)-enyl okadaate (20 microM) were as active as the free acid. The OA-induced contraction was unaffected in the presence of neomycin (5 mM), mepacrine (30 microM), 1-[N,O-bis(1,5-isoquinolinesulfonyl)-N-methyl-L tyrosyl]-4-phenylpiperaz ine (10 microM), calphostin C (3 microM) and 1-(5 isoquinolinylsulfonyl)-2-methylpiperazine (30 microM). The calmodulin inhibitor N (6-aminohexyl)-5-chloro-1-naphthalenesulfonamide hydrochloride (100 microM) did not modify the amplitude of the OA-induced contraction but significantly increased the rate of tension decay. The myosin light chain kinase inhibitor 1-(5 chloronaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine hydrochloride (1 mM) significantly reduced the peak amplitude of the contraction. Staurosporine (0.03 0.1 microM) did not modify the contractile component of the OA-induced response but inhibited the subsequent decrease in tension. In freshly dispersed myometral cells loaded with the fluorescent Ca++ indicator indo 1, OA did not produce any significant increase in [Ca++]i. OA (5- to 90-min contact) also failed to modify the intracellular levels of arachidonic acid, compared with basal values. These data suggest that in the rat uterus 1) the contractile effect of OA (20 microM) is specifically mediated by inhibition of protein phosphatases type 1 and/or 2A and is related to a direct interaction with the contractile machinery; 2) the decreasing phase of the OA-induced mechanical response could be mediated by a staurosporine-sensitive protein kinase different from protein kinase C. PMID- 9223556 TI - Selective O-desulfation produces nonanticoagulant heparin that retains pharmacological activity in the lung. AB - Heparin has potential use as an antiinflammatory treatment in many lung diseases but its therapeutic use is limited by inherent anticoagulant activity. The anticoagulant nature of heparin can be eliminated by a number of chemical treatments, but often not without loss of other important pharmacological activities. Lyophilization of porcine mucosal heparin under extreme alkaline conditions (pH > or = 13) produces a nonanticoagulant heparin remarkable for the selective loss of only 2-O and 3-O sulfates, leaving 6-O and N-sulfates intact. In contrast to the commonly used nonanticoagulant analog N-desulfated, N reacetylated heparin, selectively O-desulfated heparin retains potent activity as an inhibitor of the cationic neutrophil proteases human leukocyte elastase and cathepsin G, both in vitro and in vivo. Selectively O-desulfated heparin also inhibits complement lysis of erythrocytes, prevents ischemia-reperfusion injury of the lung, remains a potent antiproliferative treatment for cultured airway smooth muscle and normalizes altered neuronal M2 muscarinic receptor sensitivity and bronchial hyperreactivity after antigen challenge. These retained pharmacologic properties suggest possible use of this new nonanticoagulant heparin for the treatment of a variety of lung disorders. PMID- 9223557 TI - Electrophysiological characterization of the prokinetic agents cisapride and mosapride in vivo and in vitro: implications for proarrhythmic potential? AB - In the present study the electrophysiological characteristics and the proarrhythmic potential of cisapride and a structurally related drug, mosapride, were compared. In the anesthetized guinea pig, cisapride and d-sotalol (0.01-10 micromol/kg i.v., n = 6) dose-dependently prolonged the duration of the monophasic action potential recorded from the left ventricle. The maximal lengthening was 18 +/- 3.2% at 1.0 micromol/kg (mean +/- S.E.M., P < .01 vs. base line) and 19 +/- 2.5% at 10 micromol/kg (P < .001) for cisapride and d-sotalol, respectively. In contrast, mosapride did not increase this variable. In a rabbit model of the acquired long QT syndrome, infusion of cisapride (0.3 micromol/kg/min for 10 min maximum, n = 6), but not mosapride or vehicle, was associated with a significant lengthening of the QTU interval (43 +/- 3.8 ms, P < .01). Furthermore, torsades de pointes appeared in two of the six rabbits given cisapride. In isolated rabbit Purkinje fibers (PF), cisapride increased the action potential duration (48 +/- 5.6% at 0.1 micromol/l, P < .01 vs. control, n = 4). Mosapride did not significantly influence the action potential duration (3 +/- 2.0% increase at 1.0 micromol/l, n = 6). However, after mosapride was washed out, the addition of cisapride (0.1 micromol/l) caused a 46 +/- 3.2% lengthening of the action potential duration (P < .01 vs. 1.0 micromol/l mosapride). Early afterdepolarizations and triggered activity appeared in four of eight cisapride superfused PF stimulated at a very low frequency (0.1 Hz). In isolated rabbit cardiomyocytes, cisapride concentration-dependently blocked (IC50 = 9 nmol/l) the rapid component of the delayed rectifying K+ current (I(Kr)). Mosapride was approximately 1000-fold less potent in blocking I(Kr) (IC50 = 4 micromol/l). It is concluded that the electrophysiological characteristics of cisapride may explain the recently reported propensity to prolong the QT interval and to induce torsades de pointes in susceptible patients, although a structurally related benzamide, mosapride, did not appear to have electrophysiological features of relevance for induction of torsades de pointes in common with cisapride. PMID- 9223558 TI - Comparative alpha-1 adrenoceptor subtype selectivity and functional uroselectivity of alpha-1 adrenoceptor antagonists. AB - We investigated the relevance of selectivity for a given alpha-1-adrenoceptor subtype for in vivo uroselectivity of several alpha-1-adrenoceptor antagonists (alfuzosin, doxazosin, prazosin, tamsulosin, terazosin and 5-Me-urapidil). Comparison of the affinities of these alpha-1-adrenoceptor antagonists at the cloned alpha-1a, alpha-1b and alpha-1d-adrenoceptor subtypes revealed that tamsulosin and 5-Me-urapidil showed selectivity for the alpha-1a subtype. No significant correlations were found between the affinities for alpha-1b or alpha 1d-adrenoceptors and the pK(B) values obtained against phenylephrine-induced contraction of the rabbit prostate in vitro. In contrast, the antagonist potencies in rabbit prostate were correlated (r = 0.89, P < .05) with the pKi values for the alpha-1a-adrenoceptor subtype. However, the pK(B) values were consistently smaller (by 0.6 to 1.9 log unit) than the pKi values for the alpha 1a-adrenoceptor subtype, a result that suggests that the alpha-1-adrenoceptor mediating urethral contractions does not have all the characteristics of the alpha-1a-adrenoceptor. The simultaneous measurement of urethral and arterial pressures in the same conscious male rat allows evaluation of the functional uroselectivity of these antagonists based on their respective effects on both pressures. Dose ranges were selected according to effects on urethral pressure and most antagonists were found effective within the 3 to 100 microg/kg i.v. range. Alfuzosin markedly decreased urethral pressure and either did not decrease blood pressure (10-30 microg/kg) or slightly decreased it at the highest dose tested (100 microg/kg). Doxazosin did not produce sustained reductions in urethral pressure until a dose of 30 microg/kg. Blood pressure was not reduced until 100 microg/kg. Prazosin reduced urethral pressure and blood pressure within the same dose-range whereas terazosin did not decrease urethral pressure at doses that significantly decreased blood pressure (30 and 100 microg/kg). 5-Me urapidil, an alpha-1a-selective compound did not significantly modify urethral and blood pressure whereas tamsulosin, another alpha-1a-selective compound reduced urethral pressure and blood pressure within the same dose range. In conclusion, in the conscious male rat the functional uroselectivity is not correlated with a selective affinity for the alpha-1a-adrenoceptor subtype. PMID- 9223559 TI - Structure-activity relationships of spontaneous nitric oxide releasers, FK409 and its derivatives. AB - (+/-)-(E)-4-Ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (FK409) shows both potent in vitro vasorelaxant and antiplatelet activities via nitric oxide (NO) generated spontaneously from the compound. In this study, we measured spontaneous NO-releasing rates of a series of FK409 derivatives, of which chain lengths or substituents were systematically modified, in sodium-phosphate buffer solution at pH 7.4. Furthermore, we studied their in vitro antiplatelet and vasorelaxant effects to evaluate relationships between spontaneous NO-releasing activities of FK409 analogs and their biological activities. FK409 derivatives were found to possess different spontaneous NO-releasing rates and biological activities according to their structural modification. In addition, these studies revealed a close correlation between NO-releasing rates of FK409 derivatives and their in vitro antiplatelet activities in human platelet-rich plasma, whereas the in vitro vasorelaxant activities of these compounds in isolated rat aorta did not correlate with the rates of NO liberation. The vasorelaxant effects were supposed to be affected by the structural properties of FK409 derivatives as well as their NO-releasing abilities. PMID- 9223560 TI - The biodisposition and metabolism of anandamide in mice. AB - The endogenous cannabinoid anandamide (AN) has been reported to produce pharmacological effects similar to those of delta9-tetrahydrocannabinol but with a shorter duration of action. Also, AN is known to be metabolized to arachidonic acid. The purpose of this study was to examine the time course of distribution and metabolism of AN. Male mice were each administered 20 microCi 3H-AN and 50 mg/kg AN (i.v.). At 1, 5, 15 and 30 min after administration, the animals were sacrificed, and various tissues were removed, solubilized and counted to determine the distribution of 3H. Also, samples from brain, adrenal gland and plasma were extracted with ethyl acetate and analyzed by HPLC to separate 3H labeled AN, arachidonic acid and other metabolites. AN was detectable in brain by 1 min after injection. At 1 min after injection, the rank order of radioactivity per milligram or microliter of tissue was adrenal > lung > kidney > plasma > heart > liver > diaphragm > brain > fat. Although the 1 and 5 min metabolic profiles of brain 3H showed that AN was clearly present, most AN had already been transformed to arachidonic acid and other polar metabolites, and there were almost no detectable brain levels of AN at 15 and 30 min. In plasma and adrenal gland, AN was the predominant form at 1 and 5 min. Our experiments confirm that AN quickly reaches the brain and suggest that rapid metabolism of AN plays a key role in the time course of the pharmacological activity of this naturally occurring cannabinoid receptor ligand. PMID- 9223561 TI - Activation of alpha-2 adrenergic receptors inhibits norepinephrine release by a pertussis toxin-insensitive pathway independent of changes in cytosolic calcium in cultured rat sympathetic neurons. AB - Whole-cell electrophysiological studies suggest that sympathetic nerve alpha-2 adrenergic receptors are coupled to voltage-dependent N-type calcium channels through the Gi family of proteins to inhibit neurotransmitter release. Because most nerve terminals are too small for direct electrophysiological recordings, the aim of this study was to examine the relationship between alpha-2 adrenergic receptor-mediated inhibition of norepinephrine release and the rise in cytosolic calcium in neurites from cultured sympathetic neurons. In cultured rat superior cervical ganglion neurons, the alpha-2 adrenergic receptor agonists, UK-14304 (0.01-10 microM) and oxymetazoline (0.1-10 microM), and the N-type calcium channel blocker, omega-conotoxin GVIA (0.1-10 nM), inhibited the release of tritiated norepinephrine in response to electrical stimulation (1 Hz, 30 pulses, 0.1 ms, 70 V). The inhibitory effect of the alpha-2 adrenergic receptor agonists was not altered by pretreatment with pertussis toxin (200 ng/ml, 18 h), although pertussis toxin blocked the inhibition of forskolin-stimulated cAMP accumulation by UK-14304. In fura-2 loaded cells, electrical stimulation (1 Hz, 30 pulses, 0.1 ms, 70 V) increased cytosolic calcium in sympathetic neuronal processes. Blockade of N-type calcium channels with omega-conotoxin (1 and 10 nM) reduced the rise in cytosolic calcium by 25 +/- 3% and 52 +/- 6%, respectively, whereas UK-14304 and oxymetazoline did not alter the electrically stimulated rise in cytosolic calcium. These data suggest that blockade of N-type calcium channels with omega conotoxin GVIA inhibits stimulated norepinephrine release and cytosolic calcium measured with fura-2 at similar concentrations, whereas activation of alpha-2 adrenergic receptor inhibits norepinephrine release by a pathway that is insensitive to pertussis toxin and changes in cytosolic calcium in neurites from cultured rat superior cervical ganglion cells. PMID- 9223562 TI - Ethodin: pharmacological evidence of the interaction between smooth muscle and mast cells in the myometrium. AB - Ethodin has been used to induce labor through a mechanism that does not involve the estrogen-preparatory process being postulated as necessary for ensuring the events in a normal labor. The cellular mechanisms involved in that process are unknown. We used an isolated organ bath preparation for mouse uterine horns and a primary culture of mouse myometrial smooth muscle cells to analyze the cellular mechanisms involved in the contractile action of this drug in the myometrium. Ethodin at a concentration of 10 microM and Compound 48/80 (1 microg/ml) evoked contractions of uterine horns in an isolated organ bath preparation. Uterine contractile responses showed a transient increase in contractile tension that lasted 2 to 3 min. Tachyphylaxis was observed after four or five successive stimuli, which consisted in additions and washings of the drug at an interval of 10 min. The primary smooth muscle mouse myometrium cells contained a high proportion of relaxed cells that varied widely in length (5-160 microm). Cell lengths decreased in response to the application of serotonin (10 microM) and oxytocin (0.1 microM) but were not affected after the addition of ethodin (10 microM). However, the cells contracted after a purified fraction of mast cells that had been degranulated by the action of the drug ethodin, which was added to the culture medium. These results provide some evidence related to the mechanism of myometrial contractile action of ethodin and support the hypothesis that mast cells may be involved in the regulation of myometrium contractility. PMID- 9223563 TI - The potassium channel blockers 4-aminopyridine and tetraethylammonium increase the spontaneous basal release of [3H]5-hydroxytryptamine in rat hippocampal slices. AB - Previous investigations have demonstrated that compounds capable of blocking presynaptic potassium channels can stimulate neurotransmitter release at both peripheral and central synapses. This study examined the in vitro effects of the "classical" potassium channel blockers 4-aminopyridine (4-AP) and tetraethylammonium (TEA) on the spontaneous basal release of [3H]5 hydroxytryptamine ([3H]5-HT) from rat hippocampal slices using an automated superfusion apparatus. 4-AP and structural analogs increased the spontaneous basal release of [3H]5-HT in a concentration-related manner. The rank order of potencies from the estimated EC50 values indicated that 3,4-diaminopyridine (0.88 mM) approximately 4-AP (1.2 mM) > 2-AP (89 mM) > 3-AP (100 mM) > pyridine (256 mM). TEA stimulated [3H]5-HT release with an estimated EC50 value of 63 mM and was less efficacious than the pyridine congeners. The enhancement of release induced by 1 mM 4-AP was additive with 100 mM TEA and 5 microM veratridine but not with 3,4-diaminopyridine or KCl (25 and 50 mM). The release induced by 4-AP (0.3, 1 and 10 mM) and TEA (30, 100 and 300 mM) was significantly attenuated in a calcium-free buffer containing 1 mM ethylene glycol-bis(b-aminoethyl ether N,N,N',N'-tetraacetic acid. Tetrodotoxin (1 microM), a sodium channel blocker, was unable to block the response to 4-AP (1 mM) and TEA (100 mM). Notably, this concentration of tetrodotoxin reduced the stimulation of [3H]5-HT release produced by the sodium channel opener veratridine (5 microM). Taken together, the results demonstrate that potassium channel blockade can enhance the spontaneous basal release of [3H]5-HT in rat hippocampal slices. These effects are at least partly dependent on extracellular calcium and do not appear to be mediated by modulating sodium channel function. PMID- 9223564 TI - Antagonistic modulation between the delta opioid agonist BW373U86 and the mu opioid agonist fentanyl in mice. AB - This study was performed to assess the interactions that occur between delta- and mu opioid receptors by studying effects of the systemically active nonpeptide delta agonist BW373U86 and the mu agonist fentanyl in mice. Concentrations of the compounds were varied, and analgesic responses were determined by 55 degrees C hot-plate assays. BW373U86 produced hot-plate antinociceptive activity along with convulsive side effects. These effects could be antagonized by the selective delta antagonist naltrindole. Fentanyl produced hot-plate antinociceptive activity with Straub tail and hyperactivity as side effects. When BW373U86 and fentanyl were coadministered, BW373U86 convulsive activity was attenuated by fentanyl in a dose-dependent manner and the fentanyl-induced Straub tail effect was antagonized by BW373U86, also in a dose-dependent manner. Hot-plate analgesic activity was additive between the two compounds. The delta antagonist naltrindole partially antagonized the ability of BW373U86 to block the fentanyl-induced Straub tail effect. The mu antagonist beta-funaltrexamine antagonized the fentanyl-induced blockade of the convulsive effects of BW373U86. These data suggest that complex inhibitory interactions take place between mu and delta receptors in mice. Future studies are clearly needed to study the neuromodulatory effects of mu and delta receptors. The widespread use of mu agonists in the clinic indicates that a large number of patients exist who could greatly benefit from the conjunctive use of delta pharmaceuticals. PMID- 9223565 TI - Three allosteric modulators act at a common site, distinct from that of competitive antagonists, at muscarinic acetylcholine M2 receptors. AB - Functional studies were conducted on guinea pig atrial muscarinic acetylcholine M2 receptors with the allosteric modulators heptane-1,7-bis(dimethyl-3' phthalimidopropyl)ammonium bromide (C7/3'-phth), gallamine and alcuronium to determine whether these ligands are able to recognize a common accessory site. The three modulators inhibited the negative inotropic response to carbachol in this tissue. When used in combination, C7/3'-phth and gallamine or C7/3'-phth and alcuronium gave dose ratios that were either additive or underadditive. In contrast, the combinations of C7/3'-phth or alcuronium with the competitive antagonists, N-methylscopolamine or atropine, yielded supra-additive dose ratios. The data could be reconciled with a model involving a ternary complex between (1) the receptor, (2) carbachol, N-methylscopolamine or atropine acting at the orthosteric binding site and (3) C7/3'-phth, alcuronium or gallamine acting at a common, allosteric site with varying degrees of heterotropic cooperativity. PMID- 9223566 TI - EEG spectral analysis of the neuroprotective kappa opioids enadoline and PD117302. AB - The present study characterized the electroencephalographic (EEG) effects of the neuroprotective kappa opioids enadoline and PD117302 in conscious, freely moving rats with the use of computer-assisted spectral analysis (CASA). Enadoline (25 100 microg/kg) or PD117302 (1.25-5.0 mg/kg) was administered intravenously to rats implanted with cortical EEG electrodes. Although both drugs produced an immediate, mild sedation, there were no signs of head-weaving or ataxia, and there was little visual evidence of opioid-like EEG slow-wave bursts or seizures. Both drugs produced only modest increases in total EEG power that were not dose dependent. In contrast, CASA revealed significant dose-dependent frequency shifts in relative power distributions, thereby identifying distinct kappa opioid alterations in awake EEG activity; EEG power decreased in the 0- to 4-Hz frequency band with concomitant increases in power measured in the 4- to 8-Hz frequency range. The kappa opioids produced a dose-dependent consolidation of the EEG waveform centered about a peak frequency of 5.0 Hz (for enadoline) or 4.8 Hz (for PD117302) and a significant shift in the mean EEG frequency from 6.6 Hz (predrug) to 6.2 Hz (postdrug). Further CASA revealed significant postdrug decreases in the edge frequency, mobility and complexity of the EEG. Both drugs produced moderate increases in the latency to slow-wave sleep (SWS). Overall, enadoline (ED50 = 18 microg/kg) was approximately 94 times more potent than PD117302 (ED50 = 1690 microg/kg) in producing the kappa EEG profile. Because the kappa-induced EEG changes were stereospecific for the (-)-enantiomers and inhibited by norbinaltorphimine (nor-BNI), the EEG "fingerprint" described in this study could be attributed to specific activation of brain kappa opioid receptors. PMID- 9223567 TI - Role of CYP3A4 in human hepatic diltiazem N-demethylation: inhibition of CYP3A4 activity by oxidized diltiazem metabolites. AB - The antihypertensive agent diltiazem (DTZ) impairs hepatic drug metabolism by inhibition of cytochrome P450 (CYP). The accumulation of DTZ metabolites in serum occurs during prolonged therapy and leads to decreased DTZ elimination. Thus, DTZ metabolites may contribute to CYP inhibition. This study assessed the role of human CYPs in microsomal DTZ oxidation and the capacity of DTZ metabolites to inhibit specific CYP activities. DTZ N-demethylation varied 10-fold in microsomal fractions from 17 livers (0.33-3.31 nmol/mg of protein/min). DTZ oxidation was correlated with testosterone 6beta-hydroxylation (r = 0.82) and, to a lesser extent, tolbutamide hydroxylation (r = 0.59) but not with activities mediated by CYP1A2 or CYP2E1. CYP3A4 in lymphoblastoid cell microsomes catalyzed DTZ N demethylation but CYP2C8 and CYP2C9 were also active (approximately 20% and 10% of the activity supported by CYP3A4); seven other CYPs produced little or no N desmethyl DTZ from DTZ. The CYP3A4 inhibitors ketoconazole and troleandomycin decreased microsomal DTZ oxidation, but inhibitors or substrates of CYP2C, CYP2D and CYP2E1 produced no inhibition. Some inhibition was produced by alpha naphthoflavone, a chemical that inhibits CYP1As and also interacts with CYP3A4. In further experiments, the capacities of DTZ and three metabolites to modulate human CYP 1A2, 2E1, 2C9 and 3A4 activities were evaluated in vitro. DTZ and its N desmethyl and N,N-didesmethyl metabolites selectively inhibited CYP3A4 activity, whereas O-desmethyl DTZ was not inhibitory. The IC50 value of DTZ against CYP3A4 mediated testosterone 6beta-hydroxylation (substrate concentration, 50 microM) was 120 microM. The N-desmethyl (IC50 = 11 microM) and N,N-didesmethyl (IC50 = 0.6 microM) metabolites were 11 and 200 times, respectively, more potent. From kinetic studies, N-desmethyl DTZ and N,N-didesmethyl DTZ were potent competitive inhibitors of CYP3A4 (Ki = approximately 2 and 0.1 microM, respectively). CYP3A4 inhibition was enhanced when DTZ and N-desmethyl DTZ underwent biotransformation in NADPH-supplemented hepatic microsomes in vitro, supporting the contention that inhibitory metabolites may be generated in situ. These findings suggest that N demethylated metabolites of DTZ may contribute to CYP3A4 inhibition in vivo, especially under conditions in which N-desmethyl DTZ accumulates, such as during prolonged DTZ therapy. PMID- 9223568 TI - Pharmacological profile of YM087, a novel potent nonpeptide vasopressin V1A and V2 receptor antagonist, in vitro and in vivo. AB - The biochemical and pharmacological profile of YM087, 4'-[(2-methyl-1,4,5,6 tetrahydroimidazo[4,5-d][1]benzazepin- 6-yl)-carbonyl]-2-phenylbenzanilide monohydrochloride, a newly synthesized nonpeptide vasopressin (AVP) antagonist, was investigated in several in vitro and in vivo studies. YM087 showed high affinity for V1A receptors from rat liver and V2 receptors from rat kidney with Ki values of 0.48 and 3.04 nM, respectively. YM087 also inhibited [3H]oxytocin (OT) binding to rat uterus (OT receptors) plasma membranes with a Ki value of 44.4 nM, and at 100 microM did not affect the binding of [3H]AVP to anterior pituitary (V1B receptors) plasma membranes, which indicated that it had less affinity for these OT and V1B receptors. YM087 had no effect on cytosolic free calcium concentration ([Ca++]i) itself, but suppressed AVP-induced increase in [Ca++]i of cultured vascular smooth muscle cells at the same concentrations as the binding affinities. Furthermore, YM087 potently blocked AVP-induced cAMP production of cultured renal epithelium cells concentration dependently and had no agonistic activities. In in vivo studies, intravenous administration of YM087 inhibited the pressor response to exogenous AVP in pithed rats and produced an aquaretic effect in dehydrated conscious rats in a dose-dependent manner. These results demonstrate that YM087 is a potent and nonpeptide dual AVP V1A and V2 receptors antagonist and can be used in future studies to help clarify the physiological and pathophysiological roles of AVP. PMID- 9223570 TI - Ultrasonic vocalizations in rat pups: modulation at the gamma-aminobutyric acidA receptor complex and the neurosteroid recognition site. AB - Agonists acting at benzodiazepine, gamma-aminobutyric acidA, barbiturate and neurosteroid recognition sites were studied for their attenuation of separation induced ultrasonic vocalizations (USV) in rat pups. The behavioral effects of the neuroactive steroid 3alpha-hydroxy-5alpha-pregnan-20-one (allopregnanolone) were assessed when the drug was administered alone and in combination with agonists and antagonists acting at the gamma-aminobutyric acidA receptor complex. At 7 days postpartum, male and female Long-Evans rat pups were separated from the dam and littermates, and placed on a 20 degrees C surface for 2 min. Allopregnanolone (1-30 mg/kg s.c.), alprazolam (0.03-1 mg/kg s.c.), diazepam (0.1-3 mg/kg s.c.), muscimol (0.03-0.3 mg/kg s.c.) and pentobarbital (1-30 mg/kg s.c.) dose dependently decreased USV. Pretreatment with flumazenil (0.1 mg/kg s.c.) antagonized alprazolam's and diazepam's USV-suppressive effects; bicuculline (2 mg/kg s.c.) reversed muscimol's USV-suppressive effects. Allopregnanolone (3 mg/kg s.c.) produced a 4- to 7-fold leftward shift in alprazolam's and diazepam's USV-suppressive effects, and also produced a modest leftward shift in pentobarbital's USV dose-effect function. Neither flumazenil, bicuculline, nor picrotoxin (1 mg/kg s.c.) altered allopregnanolone's USV-suppressive effects. These results suggest that the USV-suppressive effects of the neurosteroid allopregnanolone are mediated at the gamma-aminobutyric acidA receptor complex, and are independent from a direct action on the benzodiazepine or gamma aminobutyric acidA recognition sites on this complex. PMID- 9223571 TI - Antagonism of N-methyl-D-aspartate receptors by sigma site ligands: potency, subtype-selectivity and mechanisms of inhibition. AB - Recent studies propose that sigma site ligands antagonize N-methyl-D-aspartate (NMDA) receptors by either direct, or indirect mechanisms of inhibition. To investigate this question further we used electrical recordings to assay actions of seventeen structurally diverse sigma site ligands on three diheteromeric subunit combinations of cloned rat NMDA receptors expressed in Xenopus oocytes: NR1a coexpressed with either NR2A, 2B or 2C. The sigma site ligands had a wide range of potency for antagonizing NMDA receptor currents. Steady-state IC50 values ranged between approximately 0.1 to >100 microM. In all cases inhibition was non-competitive with respect to glycine and glutamate. Five structurally related sigma ligands [eliprodil, haloperidol, ifenprodil, 4-phenyl-1-(4 phenylbutyl)-piperidine and trifluperidol] were strongly selective for NR1a/2B receptors. The other drugs were weakly selective or nonselective inhibitors. There was no correlation between sigma site affinity and potency of NMDA receptor antagonism for any subunit combination. Inhibition of NR1a/2B receptors by the selective antagonists was independent of voltage whereas inhibition by the weakly selective antagonists was voltage dependent. Potency of 10 sigma ligands was cross-checked on NMDA currents in cultured rat cortical neurons. There was close correspondence between the two assay systems. Our results argue that antagonism of NMDA receptor currents by the sigma ligands tested is due to direct effects on the receptor channel complex as opposed to indirect effects mediated by sigma receptors. Inhibition occurs via sites in the NMDA receptor channel pore, or via allosteric modulatory sites associated with the NR2B subunit. PMID- 9223569 TI - Extracellular Mg++ manipulation prevents the proarrhythmic activity of cromakalim in ischemic/reperfused diabetic hearts. AB - Cromakalim, an adenosine triphosphate-sensitive potassium channel opener, shows proarrhythmic activity at moderate doses (1-10 micromol/liter) in the ischemic and reperfused myocardium. We studied the effects of extracellular Mg++ ([Mg++]o) on the incidence of reperfusion-induced ventricular fibrillation and ventricular tachycardia in isolated working hearts (n = 12 in each group) subjected to 20 min of global ischemia followed by 30 min of reperfusion, a model eliciting a low incidence of reperfusion arrhythmias, obtained from 8-wk streptozotocin-induced diabetic rats. Cromakalim, at a concentration of 3 micromol/liter, perfused 5 min before the induction of ischemia and throughout reperfusion increased the incidence of ventricular fibrillation and ventricular tachycardia from their drug free diabetic control values of 25 and 42% ([Mg++]o = 1.2 mmol/liter) to 92% (P < .05) and 100% (P < .05), respectively. Glibenclamide at a concentration of 3 micromol/liter prevented the proarrhythmiac activity of cromakalim. Increasing concentration of [Mg++]o to 2.4, 3.6 and 4.8 mmol/liter in the perfusion buffer, the arrhythmogenic effect of cromakalim was also abolished. Thus, with 2.4, 3.6 and 4.8 mmol/liter of [Mg++]o perfused before the administration of cromakalim and the onset of ischemia, the incidence of reperfusion-induced ventricular tachycardia was reduced from 92% (in cromakalim treated group) to 67%, 42% (P < .05), and 25% (P < .05), respectively. The incidence of reperfusion-induced ventricular tachycardia showed the same pattern. Elevated [Mg++]o prevented the cromakalim-induced cellular Na+ gain and K+ loss, measured by atomic absorption spectrophotometer. [Mg++]o could prevent the proarrhythmic activity of cromakalim, and the use of cromakalim as an antihypertensive or antiischemic agent may be of particular concern in the population of postischemic diabetic subjects who are known to be at high risk of sudden coronary death. PMID- 9223572 TI - Potencies of leflunomide and HR325 as inhibitors of prostaglandin endoperoxide H synthase-1 and -2: comparison with nonsteroidal anti-inflammatory drugs. AB - The relative anti-inflammatory activities of the immunomodulators HR325 and leflunomide, or its active metabolite A77 1726, were examined by determining potencies in vitro on prostaglandin endoperoxide H synthase (PGHS) and in vivo in rat air pouch inflammation. Nonsteroidal anti-inflammatory drugs (NSAIDs) were used as comparators. HR325 was more potent than A77 1726 as an inhibitor of PGHS in guinea pig polymorphonuclear leukocytes (IC50 = 415 and 4400 nM, respectively) and on isolated ovine PGHS-1 (IC50 = 64 and 742 microM) and PGHS-2 (IC50 = 100 and 2766 microM). In vivo, in rat carrageenan air pouch inflammation, HR325 but not leflunomide at 25 mg/kg inhibited accumulation of leukocytes (48%) and PGE2 (61%). HR325 was also more potent than A77 1726 against human peripheral blood mononuclear cell PGHS-1 [IC50 = 1.6 and 25.6 microM (thromboxane B2 production) or 1.1 and 8 microM (PGE2 production)] and lipopolysaccharide-induced PGHS-2 in human adherent peripheral blood mononuclear cells (IC50 = 435 nM and 9.5 microM) and peripheral blood polymorphonuclear leukocytes (IC50 = 91 nM and 3.2 microM). HR325 had low PGHS-2 selectivity in the human (2.5-12-fold) and was a more potent PGHS-2 inhibitor than naproxen, ibuprofen and piroxicam (28-fold). Assays using endogenous arachidonic acid as substrate yielded IC50 values for NSAIDs that were in general markedly lower than those published for assays using 10 microM substrate. With this approach, piroxicam had reasonable activity on human PGHS-2 (IC50 = 260-290 nM). Only NS398 and flufenamic acid were PGHS-2 selective in the human (90-330-fold and 37-60-fold, respectively); the other NSAIDs were either PGHS-1-selective (naproxen, ibuprofen, flurbiprofen and indomethacin) or nonselective (piroxicam and diclofenac). Inclusion of 10% human plasma reduced HR325 potency against PGHS-1 in human peripheral blood mononuclear cells approximately 32-fold (IC50 = 36 microM). Plasma protein binding further reduced HR325 potency (IC50 = 164 microM) and minimized the difference between HR325 and A77 1726 (IC50 = 292 microM) in a whole blood PGHS assay. Whether the greater activity against human PGHS-2 would allow HR325 to exhibit NSAID-like therapeutic effects in humans remains unclear. PMID- 9223573 TI - Ventilation in morphine-maintained rhesus monkeys. I: Effects of naltrexone and abstinence-associated withdrawal. AB - The effects of naltrexone on ventilation were examined in three rhesus monkeys maintained on 3.2 mg/kg/day morphine. Before the onset of the daily morphine dosing regimen, naltrexone had only modest effects on ventilation; a dose of 32 mg/kg increased ventilatory rate in the presence of normal air to 36 +/- 1 breaths/min, from a baseline rate of 25 +/- 1 breaths/min. Naltrexone did not affect other measures of ventilation in the presence of normal air or 5% CO2. Subsequent to the onset of the daily morphine injection regimen, naltrexone dose dependently increased ventilatory rate at doses 4 orders of magnitude lower (0.001-0.01 mg/kg) than those effective in nondependent monkeys. A dose of 0.01 mg/kg naltrexone in morphine-maintained monkeys increased ventilatory rate in the presence of normal air to 52 +/- 4 breaths/min. Naltrexone also dose-dependently increased ventilatory rate in the presence of 3% and 5% CO2; tidal volume was not affected by naltrexone administration. Doubling the maintenance dose of morphine to 6.4 mg/ kg/day further increased the ventilatory effects of naltrexone. Withholding the maintenance dose of morphine also increased ventilatory rate without affecting tidal volumes, in a manner similar to that seen after naltrexone administration. These results are consistent with the view that changes in ventilation can be used to measure precipitated and abstinence associated opioid withdrawal in monkeys. PMID- 9223574 TI - Ventilation in morphine-maintained rhesus monkeys. II: Tolerance to the antinociceptive but not the ventilatory effects of morphine. AB - The antinociceptive and ventilatory effects of morphine and other opioid agonists were determined in three rhesus monkeys during a period of morphine maintenance, as well as before and after the chronic exposure to morphine. Before the onset of the daily dosing regimen, morphine increased tail-withdrawal latencies from 50 degrees C water, with an ED50 of 6.4 +/- 2.1 mg/kg. Daily injection of 3.2 mg/kg morphine produced a rightward displacement of the morphine dose-response curve, increasing the ED50 of morphine to 28.4 +/- 12.3 mg/kg. Doubling the daily morphine dose to 6.4 mg/kg resulted in a further shift to the right of the dose response curve of morphine. After cessation of the daily dosing regimen, the morphine dose-response curve for producing antinociceptive effects returned toward baseline. The antinociceptive effects of the kappa opioid agonist, ethylketazocine, were similar during the period of daily exposure to morphine, and after cessation of the daily dosing regimen. Before the onset of the daily dosing regimen, morphine, ethylketazocine, fentanyl, butorphanol and nalbuphine decreased ventilation in the presence of air or air mixed with CO2. The baseline ED50 value of morphine for decreasing minute volume in the presence of 5% CO2 was 2.9 +/- 0.8 mg/kg. The ventilatory effects of morphine and other mu opioid agonists tested were not attenuated during the daily morphine-dosing regimen. After 40 weeks of daily injections of 3.2 mg/kg morphine, the ED50 of morphine for decreasing minute volume in 5% CO2 was 2.3 +/- 1.0 mg/kg, and when the daily dose was doubled to 6.4 mg/kg morphine, the ED50 of morphine was 1.5 +/- 0.5 mg/kg. The ventilatory depressant effects of the daily injection 3.2 mg/kg morphine were also unchanged during morphine maintenance. The differential development of tolerance to the antinociceptive and ventilatory effects of morphine demonstrates a separation of these two mu opioid agonist effects in rhesus monkeys. PMID- 9223575 TI - Carvedilol retards sudden loss of contraction during early regional myocardial ischemia in feline hearts. AB - The purpose of our study was to investigate whether loss of myocardial contraction immediately after coronary occlusion was nonuniform, and if pretreatment with carvedilol, a vasodilating nonselective beta-adrenoceptor antagonist, could retard loss of contraction after coronary artery occlusion. Feline hearts were subjected to acute regional ischemia by total occlusion of the left anterior descending coronary artery. The animals were either treated with vehicle (control group) or with carvedilol 1 mg/kg i.v. before left anterior descending coronary artery occlusion (n = 9 in each group). Regional contraction in the left anterior descending coronary artery perfused region of the heart was studied by cross-oriented sonomicrometry. In control animals, circumferential (subepicardial) contraction ceased after 10 sec, whereas longitudinal (subendocardial) contraction ceased immediately after left anterior descending coronary artery occlusion. Loss of contraction in animals treated with carvedilol was significantly slower compared to controls. Circumferential contraction ceased between 30 sec and 1 min, whereas longitudinal contraction ceased after 20 sec. In conclusion, loss of contraction during the first seconds after coronary occlusion was nonuniform, with most rapid dysfunction in the subendocardium. Pretreatment with carvedilol retarded loss of contraction in both axes. PMID- 9223576 TI - Protection from gentamicin ototoxicity by iron chelators in guinea pig in vivo. AB - This study details the prevention of gentamicin-induced hearing loss in guinea pig in vivo. The approach is based on our recent demonstrations of a redox-active gentamicin-iron complex in vitro and partial attenuation of gentamicin-induced hearing loss by the iron chelators deferoxamine and 2,3-dihydroxybenzoate. In our study, guinea pigs receiving injections of gentamicin (120 mg/kg body weight daily x 19 days) developed a progressive threshold shift reaching 50 to 70 dB at 18 kHz. Concurrent treatment with different doses of 2,3-dihydroxybenzoate (30 300 mg/kg/day) reduced the threshold shift to 25 to 15 dB. Coinjection of gentamicin with dihydroxybenzoate (100 mg/kg/day) plus mannitol (15 mg/kg/day) yielded complete functional and morphological protection from gentamicin ototoxicity although partial protection was observed with combinations of dihydroxybenzoate and deferoxamine. Dihydroxybenzoate also attenuated gentamicin induced vestibular toxicity. The iron chelators and radical scavengers affected neither serum levels nor the antimicrobial efficacy of gentamicin against Escherichia coli. These results confirm that iron and free radicals play a crucial role in the toxic side effects of gentamicin. Furthermore, they suggest that iron chelators, which are well-established drugs in clinical therapy, may be promising therapeutic agents to reduce aminoglycoside ototoxicity. PMID- 9223577 TI - L-arginine deficiency causes suppression of nonadrenergic noncholinergic nerve mediated smooth muscle relaxation: role of L-citrulline recycling. AB - Studies were performed on the internal anal sphincter (IAS) smooth muscle strips obtained from opossums (Didelphis virginiana). Isometric tension and L-arginine levels of the tissues were measured under basal conditions, in the presence of electrical field stimulation (EFS) and after treatment with different concentrations of arginase. For the nonadrenergic noncholinergic nerve stimulation, short trains (4 sec) as well as continuous EFS were used. During continuous EFS, after the initial IAS relaxation, the response began to fade within several min to approximately 80% recovery of the basal tone. We also examined the influence of L-arginine and L-citrulline on these responses. For some studies, the tissues were pretreated with L-glutamine (an inhibitor of L citrulline uptake), L-glutamate or N(G)-hydroxy-L-arginine (an inhibitor of arginase). Interestingly, the basal levels of L-arginine were found to be significantly higher in the IAS (tonic smooth muscle) than in the rectal (phasic smooth muscle) smooth muscle. Arginase caused a concentration-dependent attenuation of the IAS relaxation caused by EFS. L-Citrulline and L-arginine were equipotent in reversing the attenuation. Both arginase (60 min pretreatment) and continuous EFS (tissues collected at the time of maximal recovery of the basal IAS tone after the initial relaxation) caused significant decreases in L-arginine levels. The decreases in the levels of L-arginine were restored by the exogenous administration of either L-arginine or L-citrulline. The restoration of L arginine levels by L-citrulline but not by L-arginine was selectively blocked by L-glutamine. Furthermore, the IAS relaxation, attenuated by arginase was unaffected by L-glutamine but was restored by N-hydroxy-L-arginine pretreatment. The studies suggest that L-citrulline-L-arginine recycling may play a significant role in the maintenance of IAS relaxation in response to nonadrenergic noncholinergic nerve stimulation. PMID- 9223578 TI - Basic fibroblast growth factor in a porcine model of chronic myocardial ischemia: a comparison of angiographic, echocardiographic and coronary flow parameters. AB - Recently, a number of growth factors including basic fibroblast growth factor (bFGF) have been shown to promote angiogenesis in vivo. In this study, we evaluated dose-dependent effect of bFGF administration in the setting of chronic myocardial ischemia. A total of 18 Yorkshire pigs subjected to ameroid occluder placement on the left circumflex artery were randomized to treatment with 10 (n = 6) or 100 microg (n = 5) of bFGF incorporated into heparin-alginate microspheres or inactive control pellets (n = 7). Eight weeks later, all animals underwent angiographic evaluation of collateral development as well as studies of coronary flow and global and regional left ventricular function. Both bFGF groups had significantly higher angiographic collateral index, TIMI flow scores and coronary flow in the ameroid-compromised territory compared with controls. Left ventricular function studies demonstrated improved global and regional function in both fibroblast growth factor groups with significantly better preservation of regional wall motion in high dose (100 microg) bFGF animals. We conclude that local perivascular delivery of bFGF results in significant improvement in myocardial function in the setting of chronic myocardial ischemia. PMID- 9223579 TI - Effect of albumin on the estimation, in vitro, of phenytoin Vmax and Km values: implications for clinical correlation. AB - The effect of bovine serum albumin (BSA) on human liver metabolism, in vitro, of 14C-phenytoin (PHT) was studied. Michaelis Menten parameters were determined for the conversion of PHT to p-hydroxy phenytoin in seven different microsomal preparations with the addition of 0, 2, and 4% BSA. The unbound Km (Kmu) values were 30.8 +/- 18.6, 1.57 +/- 0.21 and 1.50 +/- 0.17 microM (mean +/- S.D.), respectively; however, there was excellent agreement among the Vmax values (29.1, 31.8 and 31.5 pmol/min/mg). With intact tissue slices, BSA (4%) added to incubations of PHT had a minimal effect on the Vmax values in two of the four livers studied and resulted in a mean Kmu value of 2.20 +/- 0.59 microM, although the Kmu in the absence of BSA was 6.64 +/- 3.17. In scaling-up to the whole body, Vmax values were 3.9 and 1.0 mg/kg/day for microsomes and slices, respectively, compared to 5.9 mg/kg/day, in vivo. The Kmu values determined in the presence of albumin in both microsomes and slices were similar to those based on in vivo human steady state data (Kmu = 2-3 microM), and the intersubject variation, in vitro, was decreased in the presence of BSA. These findings for phenytoin metabolism suggest that the addition of albumin to incubation media for slices or microsome experiments may yield Km estimates that are more representative of in vivo values. PMID- 9223581 TI - Adenosine 3',5'-cyclic monophosphate-stimulated Ca++ efflux and acetylcholine release in ileal myenteric plexus are mediated by N-type Ca++ channels: inhibition by the kappa opioid receptor agonist. AB - Adenosine 3',5'-cyclic monophosphate (cAMP) is an important second messenger involved in cholinergic transmission. The aims of this study were to characterize the calcium channels associated with cyclic AMP-mediated acetylcholine release and Ca++ efflux in ileal myenteric plexus. We also examined if this process can be inhibited by agents such as opioids that inhibit N-type calcium channels via a pertussis toxin-sensitive G protein. Application of a cell permeant analogue, 8 bromoadenosine cyclic AMP (8Br-cAMP) (1 mM), and an activator of the adenylyl cyclase system, forskolin (0.1 mM), in a superfusion system resulted in both Ca++ efflux and 3H-acetylcholine (ACh) release from the dispersed myenteric ganglia. A preferential N-type Ca++ channel blocker, omega-Conotoxin GVIA (omega-CgTx, 10 100 nM), significantly inhibited 3H-ACh release stimulated by 8Br-cAMP. 10 nM omega-CgTx also totally inhibited 8Br-cAMP-induced Ca++ efflux, whereas the L type Ca++ channel blocker, nifedipine (1 microM), and the T-type Ca++ channel blocker, nickel (100 microM), both had no effects on the action of 8Br-cAMP. 3H ACh release during 0.1 mM forskolin stimulation was inhibited by pretreatment with a kappa receptor agonist, U50488H at 1 to 100 nM. In addition, U50488H significantly inhibited 3H-Ach release and Ca++ efflux elicited by 8Br-cAMP. Inhibition of 3H-ACh release by U50488H was reversed by 3 hr pretreatment with 300 ng/ml pertussis toxin. These results suggest that, in the myenteric plexus, cyclic AMP-stimulated Ca++ efflux and Ach release were mediated by N-type calcium channels. This process may be inhibited by activation of the kappa opioid receptor through pertussis toxin-sensitive G protein(s). PMID- 9223582 TI - Pharmacology of lobeline, a nicotinic receptor ligand. AB - In this study we investigated the pharmacology of lobeline, a high affinity nicotinic ligand with a unique pharmacological profile, in different in vitro and in vivo tests. Although lobeline displaced [3H]-nicotine binding sites in the rat brain with a Ki of 4.4 nM, it did not activate alpha4beta2 expressed receptors in frog oocytes. The in vivo pharmacological effects of lobeline were highly complex. Lobeline, at the time of maximal effect, dose-dependently produced motor impairment and decreased locomotor activity and body temperature in mice after s.c. treatment. However, antinociception was present after intrathecal but not after s.c. administration of lobeline in the tail-flick tests. The behavioral effects of lobeline were not blocked by pretreatment with either mecamylamine or dihydro-beta-erythroidine. In addition, lobeline given s.c. enhanced nicotine induced antinociception in a dose-related manner. No acute tolerance developed to either lobeline's behavioral or antinociceptive effect after s.c. or intrathecal administration, respectively. However, tolerance developed to lobeline's pharmacological effects after chronic treatment with the drug for 10 days (15 mg/kg, s.c. twice a day). Furthermore, cross-tolerance between lobeline and nicotine developed after chronic treatment with either drug. Although the alpha4beta2 receptor is unlikely to mediate the agonist effects of lobeline, our results indicate that lobeline does interact with the nicotinic receptor in a novel fashion. PMID- 9223580 TI - MK-801 limits neurovascular dysfunction during experimental allergic encephalomyelitis. AB - Increased permeability of the blood-brain barrier (BBB) is a characteristic of the demyelinating disease multiple sclerosis and the animal counterpart experimental allergic encephalomyelitis (EAE). In physically traumatized cerebral tissue neurovascular damage, linked with activation of the cerebroendothelial bound N-methyl-D-aspartate receptor, can be treated with the antagonist MK-801. We have examined the ability of MK-801 to modify BBB leakage and the development of disease during EAE. Prophylactic MK-801, at 0.15 mg kg(-1) body weight suppressed neurovascular breakdown, measured by a dual radioisotope technique, and significantly reduced neurological deficits (P < .05), but not perivascular lesions. A 2-fold increase in administered MK-801 completely prevented abnormal extravasation in cerebella (P < .01) and significantly inhibited BBB disruption in medulla-pons (P < .05) and cervical spinal tissues (P < .01). High-dose treatment also restricted disease development (P < .01) and lesion formation (P < .05). Therapeutic MK-801, at 0.30 mg kg(-1) body weight, completely counteracted neuroendothelial leakage in cerebella (P < .05) and inhibited BBB dysfunction in remaining tissues without restricting inflammatory cell invasion. However, doubling the dose did not further enhance suppression of neurovascular breakdown. Our use of MK-801 to control major features of EAE strongly implicates N-methyl-D aspartate receptor-dependent mechanisms in disease development and prompts consideration of a role for the receptor in the pathogenesis of human demyelinating conditions. PMID- 9223583 TI - CCD-3693: an orally bioavailable analog of the endogenous neuroactive steroid, pregnanolone, demonstrates potent sedative hypnotic actions in the rat. AB - An endogenous neuroactive steroid, pregnanolone, and an orally available synthetic analog, CCD-3693, were administered to rats at the middle of their circadian activity phase (6 hr after lights off). Electroencephalogram-defined sleep-wake states, locomotor activity and body temperature were concurrently measured 30 hr before and after treatment. Identical procedures were used to test triazolam and zolpidem. Triazolam (0.1-1.6 mg/kg), zolpidem (2.5-10 mg/kg) and the neuroactive steroids (10-30 mg/kg) produced dose-dependent increases in non rapid eye movement (NREM) sleep. At this dose and time of day (in which the rats were predominantly awake during the 6 hr before treatment) the neuroactive steroids appeared more intrinsically efficacious in promoting NREM sleep than the benzodiazepine ligands. The neurosteroids did not, however, significantly interfere with rapid eye movement sleep and were more selective in reducing (EEG) wakefulness, with relatively less locomotor activity impairment during waking than triazolam and zolpidem. In addition, the benzodiazepine receptor ligands showed distinct "rebound" wakefulness after the NREM sleep-promoting effect subsided, although the neuroactive steroids did not. In addition, in vitro binding studies and in vivo pharmacological data confirmed that CCD-3693 was orally active in standard tests of anxiety, anticonvulsant, loss-of-righting and passive avoidance. PMID- 9223585 TI - p-Glycoprotein-mediated transport of a fluorescent rapamycin derivative in renal proximal tubule. AB - The transport of a fluorescent rapamycin derivative was measured in killifish (Fundulus heteroclitus) renal proximal tubules by means of confocal microscopy and image analysis. Renal cells and tubular lumens rapidly accumulated the rapamycin analog from the medium and attained steady state within 60 min. At steady state, luminal fluorescence intensity was two to four times higher than cellular fluorescence. Cellular fluorescence intensity was a linear function of medium substrate concentration and was not affected by any treatment used. In contrast, luminal fluorescence exhibited a saturable component as the medium concentration of the rapamycin derivative was increased. Secretion into the lumen was blocked by KCN, rapamycin, cyclosporin A and substrates for p-glycoprotein (verapamil, PSC-833 and FK506), but not by substrates for the renal organic anion or organic cation transport systems, such as p-aminohippurate, leukotriene C4 or tetraethylammonium. Finally, rapamycin blocked p-glycoprotein-mediated secretion of a fluorescent cyclosporin A derivative. The data are consistent with the fluorescent rapamycin analog entering proximal tubule cells by simple diffusion and then being pumped into the tubular lumen by p-glycoprotein. They suggest that the parent compound, rapamycin, would be handled similarly. PMID- 9223584 TI - Antinociceptive profile of 3-alpha-tropanyl 2-(4-Cl-phenoxy)butyrate (SM-21) [corrected]: a novel analgesic with a presynaptic cholinergic mechanism of action. AB - The antinociceptive effect of (+/-)-3-alpha-tropanyl 2-(4-Cl-phenoxy)butyrate [corrected] (SM-21) (10-40 mg kg(-1) s.c., 10-30 mg kg(-1) i.p., 20-60 mg kg(-1) p.o., 3-20 mg kg(-1) i.v. and 5-20 microg per mouse i.c.v.) was examined in rodents and guinea pigs by using the hot-plate, abdominal constriction, tail flick and paw-pressure tests. The antinociception produced by (+/-)-SM-21 was prevented by atropine, pirenzepine and hemicholinium-3 but not by quinpirole, R (alpha)-methylhistamine, [1-[2(methylsufonyl)amino]ethyl]-4-piperidinyl]methyl-5 floro++ +-2-methoxy-1H-indole-3-carboxylate hydrochloride, N6 cyclopentyladenosine, 1-(2-methoxyphenyl)-4-[4-(2-phthalimido)butyl]piperazine hydrobromide, naloxone, 3-aminopropyl-diethoxy-methyl-phosphinic acid or reserpine. On the basis of the above data, it can be postulated that (+/-)-SM-21 exerted an antinociceptive effect mediated by a central potentiation of cholinergic transmission. Affinity profiles of (+/-)-SM-21 for muscarinic receptor subtypes, determined by functional studies (rabbit vas deferens for M1, guinea pig atrium for M2, guinea pig ileum for M3 and immature guinea pig uterus for putative M4) have shown a selectivity ratio M2/M1 of 4.6 that, although very low, might be responsible for the antinociception induced by (+/-)-SM-21 through an increase in ACh extracellular levels. In the antinociceptive dose range, (+/-) SM-21 did not impair mouse performance evaluated by the rota-rod and hole-board tests. PMID- 9223586 TI - [125I]IPH, an epibatidine analog, binds with high affinity to neuronal nicotinic cholinergic receptors. AB - An analog of epibatidine (EB) was synthesized with an iodine atom in the 2 position of the pyridyl ring. This analog, (+/-)-exo-2-(2-iodo-5-pyridyl)-7 azabicyclo[2.2.1]heptane (IPH), as well as its two stereoisomers, displayed high affinity for neuronal nicotinic receptors; therefore, radioiodinated IPH, [125I]IPH, was synthesized with specific radioactivities consistently > 1000 Ci/mmol, and its properties as a radioligand for neuronal nicotinic receptors were evaluated. The characteristics of [125I]IPH binding in tissue homogenates appeared to be virtually identical to those reported for [3H]epibatidine binding; but the high specific radioactivity of [125I]IPH greatly facilitated measurements of nicotinic receptors in tissues with relatively low receptor densities and/or where tissues are in limited supply. Autoradiography with [125I]IPH provided clear localization of nicotinic receptors in brain and adrenal gland after film exposure times of < or = 2 days. We conclude that [125I]IPH will be a very useful radioligand for the study of neuronal nicotinic receptors in brain and in peripheral ganglia. PMID- 9223588 TI - The natural product hymenialdisine inhibits interleukin-8 production in U937 cells by inhibition of nuclear factor-kappaB. AB - The nuclear factor-kappaB (NF-kappaB) family of transcription factors have been implicated in the inducible expression of genes involved in inflammatory and immune responses. As such, a specific inhibitor of NF-kappaB would be a useful therapeutic agent in a variety of inflammatory disorders. The marine natural product hymenialdisine was evaluated as an inhibitor of NF-kappaB in U937 cells. U937 cells were transfected with either a luciferase reporter plasmid containing the human immunodeficiency virus long terminal repeat or the interleukin-8 (IL-8) core promoter, both of which are activated by NF-kappaB. Hymenialdisine caused a concentration-dependent decrease in luciferase production from both reporters when the cells were stimulated with tumor necrosis factor-alpha, lipopolysaccharide or phorbol myristate acetate. An electrophoretic mobility shift assay confirmed its activity by inhibiting DNA binding of NF-kappaB. Hymenialdisine was shown to be a selective inhibitor of NF-kappaB in that it had no effect on the binding of other transcription factors to their DNA concensus motifs; these included activator protein-1, CCAAT/enhancer binding protein and Sp1. Functional studies showed hymenialdisine to be an inhibitor of IL-8 production and IL-8 mRNA formation in the U937 cell. Investigation into the mechanism of action of hymenialdisine showed that it was not due to inhibition of protein kinase C because the selective protein kinase C inhibitor RO 32-0432 was inactive against tumor necrosis factor-alpha-stimulated luciferase and IL-8 production. The compound also had no effect on IkappaB alpha or IkappaB beta phosphorylation and degradation. Thus, hymenialdisine is a potent inhibitor of NF kappaB and IL-8 production in U937 cells. PMID- 9223587 TI - Induction of prostaglandin endoperoxide synthase-2 by serine-threonine phosphatase inhibition. AB - Regulation of prostaglandin endoperoxide synthase-2 (PGHS-2) mRNA levels by serine-threonine phosphatases was examined in murine fibrosarcoma methylcholanthrene-101 cells. Okadaic acid (OA), a serine-threonine phosphatase inhibitor, induced PGE2 production and a significant increase in PGHS-2 immunoreactive protein. A specific PGHS-2 inhibitor, N-(2-cyclohexyloxy-4 nitrophenyl) methanesulphonamide, completely abolished the OA-mediated increase in PGE2 production, which suggests that the PGE2 formed in response to OA was derived from PGHS-2. OA-mediated PGHS-2 mRNA accumulation was observed at 1 hr, remained elevated for 24 hr and was blocked by actinomycin D, which indicates that OA increases PGHS-2 gene transcription. A significant post-transcriptional mechanism also contributed to the increased PGHS-2 mRNA accumulation, because the mRNA half-life was approximately 4 to 5 h in OA-stimulated cells. Tumor necrosis factor-alpha, but not OA, activated transcription factor nuclear factor-kappaB in methylcholanthrene-101 cells, as demonstrated by translocation of the nuclear factor-kappaB complex to the nucleus and disappearance of the cytoplasmic inhibitory protein, IkappaB-alpha. We conclude that inhibition of serine threonine phosphatases contributes to the up-regulation of PGHS-2 expression in an NF-kappaB-independent manner. PMID- 9223589 TI - N-(3-lodoprop-2E-enyl)-2beta-carbomethoxy-3beta-(3',4'-dichloro phenyl)nortropane (beta-CDIT), a tropane derivative: pharmacological characterization as a specific ligand for the dopamine transporter in the rodent brain. AB - N-(3-Iodoprop-2E-enyl)-2beta-carbomethoxy-3beta-(3',4'-dichl orophenyl)nortropane (beta-CDIT), a new iodinated tropane derivative, has been synthesized and radiolabeled with iodine. [125I]beta-CDIT was tested in vitro and ex vivo as a probe for the dopamine transporter site in the rat brain, and behavioral studies were performed in mice. Saturation studies in the striatum revealed that [125I]beta-CDIT bound to a single high-affinity site. The Kd value was 0.18 +/- 0.07 nM, and the corresponding Bmax value was 500 +/- 80 fmol/mg of protein. The pharmacological profile of specific [125I]beta-CDIT binding in the striatum was consistent with that of the dopamine transporter. In addition, competition studies in cerebral cortex regions with [3H]paroxetine and [3H]nisoxetine showed a very low affinity of beta-CDIT for the 5-hydroxytryptamine (Ki = 50 nM) and norepinephrine (Ki = 500 nM) transporters compared with beta-CIT (corresponding Ki values were 3 and 80 nM). In contrast, the competition of beta-CDIT with [3H]GBR 12935 in the striatal region (Ki = 29 nM) was of the same order of value as for beta-CIT (Ki = 27.5 nM). Behavioral experiments in mice showed that both beta-CDIT and beta-CIT induced stimulation of locomotor activity. Ex vivo autoradiographic studies in rats using [125I]beta-CDIT demonstrated high densities of [125I]beta-CDIT binding sites in areas known to be rich in dopaminergic innervation. Because of its high affinity and high selectivity for the dopamine transporter, [125I]beta-CDIT should be a valuable ligand for the exploration of the dopamine transporter with single-photon emission computed tomography. PMID- 9223591 TI - Contractile action of ethanol in guinea pig gastric smooth muscle: inhibition by tyrosine kinase inhibitors and comparison with the contractile action of epidermal growth factor-urogastrone. AB - We observed a contractile action of ethanol (20-500 mM) and other alcohols (methanol and propanol, but not butanol) in guinea pig gastric longitudinal (LM) and circular (CM) smooth muscle preparations. The potency order for the alcohols in the LM preparation was: ethanol = propanol > methanol; and in the CM preparation, propanol > ethanol > methanol. Like epidermal growth factor urogastrone (EGF), the contractile actions of ethanol in the LM and CM preparations required extracellular calcium and were blocked by the tyrosine kinase inhibitors, genistein and tyrphostin-47 (AG213). The tyrosine phosphatase inhibitor, pervanadate, potentiated the contractile action of ethanol in the LM preparation. Ethanol-induced contractions in both preparations were not affected by 4-methyl pyrazole, an inhibitor of alcohol dehydrogenase, and were unaffected by tetrodotoxin, atropine, prazosine or yohimbine. In the LM preparation, like EGF, the contractile action of ethanol was blocked by the cyclooxygenase inhibitor, indomethacin, and the diacylglycerol lipase inhibitor, U57,908; in the CM preparation, contractions caused by ethanol and EGF were still observed in the presence of these two inhibitors. Contractions caused by ethanol and EGF in the LM preparation were not affected by the epoxygenase inhibitor, ketoconazole; the lipoxygenase inhibitor, nordihydroguaiaretic acid; or the phospholipase A2 inhibitor, mepacrine. In contrast, in the LM preparation, EGF-induced contractions were attentuated by the EGF receptor-kinase inhibitor, PD153035; the MAP-kinase-kinase (MEK) inhibitor, PD98059; the kinase C inhibitor, GF109203X; and the phosphatidylinositol 3'-kinase inhibitors, Wortmannin and LY294002; whereas ethanol-induced contractions were unaffected by these inhibitors. Both ethanol and EGF caused small increases in the phosphotyrosyl protein content of the gastric tissue. We conclude that ethanol causes its contractile effects in the distinct gastric LM and CM preparations independent of nerve-released agonists and via a tyrosine kinase inhibitor-sensitive signal pathway that is in many respects similar to, but distinct from the one activated by EGF. PMID- 9223590 TI - Impaired contractile response to beta adrenoceptor stimulation in diabetic rat hearts: alterations in beta adrenoceptors-G protein-adenylate cyclase system and phospholamban phosphorylation. AB - The aim of this study was to explore the cellular mechanisms underlying the impaired contractile response to beta adrenoceptor stimulation in diabetic hearts. Chronic diabetes was induced in rats by a streptozotocin injection. Four to six weeks later, papillary muscles isolated from diabetic hearts exhibited marked reductions in the positive inotropic responses to isoproterenol, norepinephrine and epinephrine. The contractile responses to forskolin, 3 isobutyl-1-methylxanthine and dibutylic cyclic AMP were also prominently depressed. The density of beta adrenoceptors was decreased by 50%. However, competitive binding studies with isoproterenol showed no difference in the proportion of beta adrenoceptors with high-affinity binding between control and diabetic myocardial membranes. Determination of the levels of the alpha subunits of Gs and Gi by immunoblotting revealed markedly less expression of Gi in diabetic myocardium. The abilities of isoproterenol, sodium fluoride, 5'-guanylyl imidodiphosphate and forskolin to stimulate adenylate cyclase were preserved well in membranes prepared from diabetic hearts. Nevertheless, neither stimulation of beta adrenoceptors with isoproterenol nor direct activation of adenylate cyclase with forskolin evoked any significant increase in the degree of phosphorylation of phospholamban in diabetic hearts. These results suggest that impaired contractile response to beta adrenoceptor stimulation is not caused by an alteration in the beta adrenoceptors-Gs-adenylate cyclase system, but is possibly caused by an alteration in cellular function beyond the step of adenylate cyclase activation. PMID- 9223592 TI - Involvement of phospholipase C-gamma2 in activation of mitogen-activated protein kinase and phospholipase A2 by zooxanthellatoxin-A in rabbit platelets. AB - Zooxanthellatoxin-A (ZT-A), a polyhydroxypolyene isolated from a symbiotic dinoflagellate Symbiodinium sp., caused thromboxane A2-(TXA2) dependent and genistein-sensitive aggregation in rabbit platelets. Our study was performed to clarify the mechanism of the action of ZT-A. ZT-A caused an increase in tyrosine phosphorylation of 42-kDa protein, which is defined as p42 mitogen-activated protein kinase (MAPK) by immunoprecipitation. Although indomethacin (10 microM) completely inhibited ZT-A-induced TXB2 release, it partially inhibited the MAPK activation. The remained MAPK activation was completely inhibited by genistein (50 microM). Genistein (50 microM), by itself, abolished TXB2 release induced by ZT-A. ZT-A (2 microM) stimulated liberation of arachidonic acid and the subsequent metabolites such as TXB2 and 12-hydroperoxyeicosatetraenoic acid. However, ZT-A-stimulated phosphoinositide hydrolysis which was due to an increase in tyrosine phosphorylation of phospholipase C-(PLC)gamma2. The phosphorylation of PLC-gamma2 and the phosphoinositide hydrolysis were also partially inhibited by indomethacin (10 microM), and were abolished by a combined treatment of indomethacin (10 microM) and genistein (50 microM). ZT-A- (2 microM) induced MAPK activation in the presence of indomethacin (10 microM) was concentration dependently inhibited by staurosporine and calphostin C, protein kinase C inhibitors. PD98059 (50 microM), a MAPK kinase inhibitor, also inhibited ZT-A induced TXB2 release. Depletion of external Ca++ abolished ZT-A- (2 microM) induced MAPK activation, phosphoinositide hydrolysis, arachidonic acid liberation and TXB2 release. These results suggest that ZT-A stimulates a protein tyrosine kinase in the presence of external Ca++, resulting in the activation of MAPK probably via PLC-gamma2 and protein kinase C. The MAPK stimulated a liberation of arachidonic acid that is rapidly converted to TXA2. The released TXA2 causes aggregation accompanied with second stimulation of MAPK cascade. PMID- 9223594 TI - Effects of lysophosphatidic acid on primary cultured chick neurons. AB - Lysophosphatidic acid caused growth cone collapse in primary cultured chick neurons. This action was dose dependent and the potency was almost identical in three different neuron types, dorsal root ganglion neurons, retinal neurons, and sympathetic ganglion cells. Fifty percent of growth cones were collapsed by 10( 6) M lysophosphatidic acid. The growth cone collapse started within 2 min after lysophosphatidic acid exposure and no homologous desensitization was observed. However, this action was reversible and not toxic to the neurons. Suramin, known as an antagonist to lysophosphatidic acid, itself had growth cone collapsing activity against cultured primary neurons. PMID- 9223593 TI - Differential regulation of human antigen-specific Th1 and Th2 lymphocyte responses by isozyme selective cyclic nucleotide phosphodiesterase inhibitors. AB - Our study explores the relative efficacy of phosphodiesterase (PDE) inhibitors on antigen-specific Th1 and Th2 clonal responses. Proliferative responses for both phenotypes were down-regulated by the PDE4 inhibitor, rolipram, but not the PDE3 inhibitor, siguazodan. The Th2 clones were more sensitive than the Th1 clones to PDE4 inhibition (P < .05 at 10 and 100 microM rolipram). The addition of 1 microM of the adenylyl cyclase activator, isoproterenol, significantly decreased both the EC50 and IC50 of rolipram in both phenotypes (P < .05). Gene expression for interleukin-4, interleukin-5, or interferon-gamma, assessed by reverse transcription-polymerase chain reaction, was down-regulated by the PDE4 inhibitor, but not the PDE3 inhibitor, in each respective clone. Cytokine protein secretion paralleled the results of reverse transcription-polymerase chain reaction for IL-4 and interferon-gamma (P < .01 for each). No differential efficacy on cytokine generation parameters between T helper phenotypes was apparent. Rolipram treatment significantly elevated intracellular cyclic AMP (adenosine 3',5'-cyclic monophosphate) in clonal T cells (P < .01 for Th1 or Th2 clones); these elevations were consistently greater in the Th2 clones (P < .05). Finally, Th1 cells showed reduced gene expression for the PDE4C isoform and a lack of gene expression for the PDE4D isoform by reverse transcription-polymerase chain reaction, compared to the Th2 cells. These data demonstrate the potent immunomodulatory efficacy of PDE4 inhibition on antigen-specific T cell clones. The enhanced sensitivity of Th2 cells to PDE4 inhibition may be due, in part, to the differential expression of PDE4 isoforms between Th1 and Th2 cells. PMID- 9223595 TI - Developmentally regulated stabilization of neuronal intermediate filaments in rat cerebral cortex. AB - The expression and Triton X-100 (Triton) solubility of neuronal intermediate filament proteins were determined in the developing rat cerebral cortex. The level of expression of alpha-internexin was unchanged from embryonic day 15 (E15) to postnatal day 15 (P15), whereas expression of the mid-sized neurofilament subunit increased continually during this interval concomitant with a reduction in Triton solubility of the two proteins. The low molecular weight neurofilament subunit, first barely detected at P2, was largely insoluble in Triton from the initial time point that its solubility could be assayed, at P5, to P15. Similar expression patterns and Triton solubility profiles were obtained for neuronal intermediate filament proteins in cultured neurons from E15 cerebral cortex. These results suggest that alpha-internexin is expressed earlier than neurofilament proteins to provide a more plastic network in the early developing brain. The incorporation of neurofilament proteins apparently results in the formation of the more stable intermediate filament network found in mature neurons. PMID- 9223596 TI - Immunocytochemical localization of muscarinic m2 receptor in the rat spinal cord. AB - Muscarinic receptors have been implied in the regulation of spinal cord functions. The m2 subtype has been found to be one of the major muscarinic receptors expressed in the spinal cord. In order to determine the precise cellular localization of m2 receptor in the rat spinal cord, immunocytochemistry was performed using a commercially available specific antibody. In the dorsal horn, strong m2 immunoreactivity was detected in the substantia gelatinosa. In the ventral horn, m2 immunoreactivity was detected in patches that were associated with cell groups of motor neurons. At higher magnification, m2 immunoreactivity was detected in neuronal processes and many of them were seen in close apposition to m2-negative perikarya and proximal dendrites of motor neurons. These results indicate that m2 receptor immunoreactivity is localized in different neuronal elements of the spinal cord. PMID- 9223597 TI - Cerebrospinal fluid apolipoprotein E (apoE) levels in Alzheimer's disease patients are increased at follow up and show a correlation with levels of tau protein. AB - Apolipoprotein E (apoE) levels were compared in cerebrospinal fluid (CSF) taken on two occasions, with an average 15 months follow up, from groups of patients with Alzheimer's disease (AD: n = 18), mild cognitive impairment (MCI; n = 9) and other dementia disorders (ODD; n = 9). In these groups, CSF apoE levels were between 2-3-fold higher than values for a group of 27 healthy age-matched controls. CSF apoE levels in the AD group were significantly increased at follow up, compared to levels obtained on the first sampling occasion. For the same cases it had been shown previously that CSF tau protein levels were increased at follow up [Blomberg, M., Jensen, M., Basun, H., Lannfelt, L. and Wahlund, L-O., Neurosci. Lett., 214 (1996) 163-166]. The AD, but not MCI, ODD or control groups, also showed statistically significant correlations between CSF apoE and tau protein levels at both the first (r = 0.585, P < 0.01) and follow up (r = 0.695, P > 0.001 ) samplings. It is concluded that CSF measures of both apoE and tau may reflect an intimate relationship between these two proteins in AD and could prove useful in monitoring the progression of this condition. PMID- 9223598 TI - The histamine H1-antagonist chlorpheniramine facilitates learning in aged rats. AB - The effect of the histamine H1-receptor antagonist chlorpheniramine on inhibitory avoidance conditioning was investigated in 31-month-old rats, using a one-trial step-through avoidance task. Immediately after the learning trial, old rats were injected intraperitoneally with 5 or 10 mg/kg d-chlorpheniramine. Control groups included vehicle-injected old and adult (4-month-old) rats and a group of aged animals given an injection of 10 mg/kg chlorpheniramine 5 h after the training trial. When tested 24 h after training, aged rats receiving 10 mg/kg chlorpheniramine exhibited longer step-through latencies than vehicle-treated old controls, indicative of superior learning of the task. The hypermnestic effects of 10 mg/kg chlorpheniramine were no longer evident when injection was performed 5 h, rather than immediately after the learning trial, ruling out enduring proactive effects of the treatment on test performance. Furthermore, vehicle treated old rats showed poorer inhibitory avoidance performance than vehicle treated adult controls. Thus, the improvement in performance after the 10 mg/kg dose of chlorpheniramine can be interpreted in terms of a compensation of performance deficits in the old rats. PMID- 9223599 TI - Identification of differentially expressed mRNAs during rat C6 glial cell differentiation by mRNA fingerprinting using arbitrarily primed PCR (RAP). AB - Differentiation of glial cells is controlled by a complex program of differential expressions of many genes. To identify differentially expressed genes that are involved in rat C6 glial cell differentiation induced by dibutyryl cyclic AMP and theophylline, mRNA fingerprinting using arbitrarily primed PCR (RAP) was used. Four cDNA fragments, that were differentially expressed during differentiation, were isolated. Sequence analysis revealed that one of them, abundantly expressed during differentiation, was homologous to a hamster calcium-dependent serine protease. Another one was highly similar to rabbit dystrobrevin and the other two clones were identical to rat triose phosphate isomerase and calnexin. The results obtained suggest that the expressions of particular genes were changed and that RAP is a useful method to identify genes which are differentially expressed during glial cell differentiation. PMID- 9223600 TI - C-type natriuretic peptide suppresses arginine-vasopressin secretion from dissociated magnocellular neurons in newborn rat supraoptic nucleus. AB - Central administration of C-type natriuretic peptide (CNP) affects various neuroendocrine systems. In the present study, we examined whether CNP acts directly on arginine-vasopressin (AVP) secretion from rat supraoptic nucleus (SON) neurons, using acute dissociated cell preparations. CNP inhibited the basal secretion of AVP in a dose-dependent manner (10(-11)-10(-6) M). A- type natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) also suppressed the basal secretion of AVP, however, the effects were two-orders of magnitude less potent than CNP. CNP also suppressed All-induced AVP secretion, however, the inhibitory effect of CNP was less than that of ANP or BNP. These findings suggest that CNP inhibits the basal secretion of AVP through natriuretic peptide receptor (NPR)-B and has a role in the body water and electrolyte homeostasis in the central nervous system. PMID- 9223601 TI - Renal R2 chemoreceptor activity is attenuated after back heating in the rat. AB - Recent study in our laboratory has found that renal afferent nervous activity (RANA) was decreased during and after 42 degrees C back heating (BH). To investigate which renal sensory receptor is influenced during and after BH, a C shaped glass heating pad (42 degrees C) was used on the skin of the back overlying the kidneys. A single-unit recording was used to identify four types of renal sensory receptors, the R2 chemoreceptor (CR2), arterial mechanoreceptor (MRa), ureteropelvic mechanoreceptor (MRu) and venous mechanoreceptor (MRv) in anesthetized female Wistar rats. Renal cortical microvascular blood flow (CMBF) and urinary water, potassium and sodium output were measured. It was found that CR2 activity was significantly decreased during and after BH, but three types of MRs were not altered. CMBF and urine output were significantly increased during and after BH. It is concluded that the increase in renal hemodynamics by BH may dilute some chemicals in the kidney and decrease the firing rate of R2 chemoreceptors. PMID- 9223602 TI - Role of nitric oxide and serotonin in modulation of the cardiovascular defence response evoked by stimulation in the periaqueductal grey matter in rats. AB - In rats anaesthetised with alphaxalone/alphadolone, electrical stimulation (10 s trains of 1 ms pulses at 80 Hz, 40-80 microA) in the dorsolateral and lateral periaqueductal grey matter (PAG), the midbrain defence area, evoked a pressor response with tachycardia and vasodilatation in the hindlimb. Microinjection of 200 nl 0.66 mM 5HT, but not 200 nl 165 mM NaCl, at the site of stimulation attenuated the components of the PAG-evoked response by 75-98%. The effect of 5HT was significantly reduced by prior intracerebroventricular injection of 100 microg N-nitro-L-arginine methyl ester (an inhibitor of nitric oxide synthase) but not N-nitro-D-arginine methyl ester. Resting cardiovascular parameters did not change significantly following any of these manipulations. The results suggest that serotonin exerts an inhibitory modulation on the excitability of the midbrain defence area by a mechanism which involves nitric oxide. PMID- 9223603 TI - Routes of NMDA- and K(+)-stimulated calcium entry in rat cerebellar granule cells. AB - The routes of Ca2+ entry in response to N-methyl-D-aspartate (NMDA) and K+ depolarisation in cerebellar granule cells have been investigated using fura-2 fluorescence to measure intracellular Ca2+ concentrations ([Ca2+]i) and Mn2+ quench of fura-2 fluorescence as an index of Ca2+ entry. Removal of extracellular Na+ did not affect the [Ca2+]i elevation or the rate of Mn2+ quench of fura-2 fluorescence in response to NMDA (100 microM). K+ (25 mM) produced a [Ca2+]i increase which showed a 27% reduction in the presence of the NMDA channel blocker MK-801 (10 microM), whereas no reduction was detected in 50 mM K+ stimulated [Ca2+]i increases. K+ (25 and 50 mM)-stimulated Mn2+ quench rates were not significantly reduced by MK-801. These results demonstrate that NMDA primarily stimulates Ca2+ entry directly through the NMDA receptor without a major component of Ca2+ entry through voltage-gated Ca2+ channels (VGCCs). Under conditions which minimise the accumulation of endogenous glutamate, K+ depolarisation elicits a Ca2+ influx resulting mainly from activation of VGCCs. Additionally, these results show Mn2+ quench of fura-2 fluorescence to be a sensitive and definitive assay of Ca2+ entry through the NMDA receptor and VGCCs. PMID- 9223604 TI - Tityustoxin-induced release of ATP from rat brain cortical synaptosomes. AB - Tityustoxin, a scorpion toxin that alters the Na+ channel activity, induces release of ATP from rat brain cortical synaptosomes. The effect of tityustoxin is dependent on its concentration and incubation time. Continuously or cumulative release of ATP evoked by tityustoxin was calcium-dependent and interestingly only partially inhibited by tetrodotoxin. We suggest that tityustoxin mainly releases ATP from the vesicular pool but other pools may also be involved. PMID- 9223605 TI - Photoperiod does not act on the suprachiasmatic nucleus photosensitive phase through the endogenous melatonin, in the Syrian hamster. AB - The duration of the sensitive phase to light of the suprachiasmatic nuclei, in terms of Fos protein expression, depends on the photoperiod. In Syrian hamsters, a 4 h lengthening of the photosensitive phase occurs within 3-4 weeks after a transfer from a long to a short photoperiod. The absence of endogenous melatonin following pinealectomy does not prevent the lengthening of the photosensitive phase. Thus, even if the pineal production is able to convey photoperiodic information, it does not feed back on the circadian clock to allow its integration. PMID- 9223606 TI - Chronic cocaine intoxication alters hippocampal sodium channel function. AB - Repeated daily administration of subconvulsive doses of cocaine results in the appearance and increase in convulsive responsiveness to the drug and its lethal effects. The mechanisms involved in this increased susceptibility to cocaine induced seizure are yet unknown. In this study, we used whole cell patch-clamp recording techniques to examine the functional changes in voltage-dependent Na+ channels produced by subconvulsive doses of cocaine (45 mg/kg per day, i.p.) in rat hippocampal CA1 pyramidal neurons. Intact animals were injected with cocaine for 5-6 days. Acutely dissociated hippocampal neurons were then recorded in vitro. Our results show that an augmentation of peak Na+ currents and a shift in depolarizing direction of the steady-state inactivation were present in neurons from drug-treated rats. These changes, by making a larger proportion of Na+ channels available for opening, could increase the excitability of CA1 neurons and may contribute to the increase in convulsive responsiveness to cocaine. PMID- 9223608 TI - Dopamine D4 receptor in human peripheral blood lymphocytes: a radioligand binding assay study. AB - The expression of dopamine D4 receptor was investigated in human peripheral blood lymphocytes with a radioligand binding assay technique, using [3H]clozapine as radioligand. [3H]Clozapine was specifically bound to human peripheral blood lymphocytes. The binding was time-, temperature-, and concentration-dependent and of high affinity, with a dissociation constant (K(d)) value of 0.34 +/- 0.02 nM and a maximum density of binding sites (B(max)) value of 27 +/- 1.4 fmol/10(6) cells. The pharmacological profile of [3H]clozapine binding to human peripheral blood lymphocytes was similar to that found in Chinese hamster ovary (CHO) cells transfected with the D4 clone (D4.2 variant). The above results are consistent with molecular biology studies demonstrating the expression of a dopamine D4 receptor in immune cells and in human peripheral blood lymphocytes. The availability of a rapid and sensitive radioligand binding assay technique for the dopamine D4 receptor in human peripheral blood lymphocytes may contribute to better define the role of this dopamine receptor subtype in neurological and psychiatric disorders. PMID- 9223607 TI - Age-related decreases in mRNA for brain nuclear receptors and target genes are reversed by retinoic acid treatment. AB - Ageing is accompanied by certain problems resulting from changes of hormonal status, in particular thyroid hormone (T3) status and vitamin A status. Since retinoic acid (RA), the active metabolite of vitamin A, and T3 play physiological roles in the adult brain, the effect of ageing on the amounts of mRNA for retinoic acid (RAR and RXR) and triiodothyronine (TR) nuclear receptors were studied. Also, the expression of RA and T3 target genes, tissue transglutaminase (tTG) and neurogranin (RC3), was measured in the whole brain and in the hippocampus of mice. Relative to young (3 months) mice, aged (22 months) mice exhibited lower amounts of RAR, RXR and TR mRNA concomitantly with a lower expression of tTG and RC3. RA administration to old mice (24 h before sacrifice) was able to restore the amount of mRNA of nuclear receptors and of RC3. It is hypothesized that a decrease in the cellular action of RA and T3 could play a role, via a decrease in the expression of RC3, in the alteration of synaptic plasticity occurring in aged mice. PMID- 9223609 TI - Tiagabine exerts an anti-epileptogenic effect in amygdala kindling epileptogenesis in the rat. AB - Tiagabine (TGB) is a novel antiepileptic drug whose anticonvulsant effects are due to inhibition of gamma-aminobutyric acid (GABA) transport mediated by the GABA transporter-1. We have previously shown that TGB is effective in acute amygdala kindled seizures, and consequently we wanted to test the hypothesis that TGB also could suppress the development of kindling epileptogenesis. Rats had stereotaxically implanted stimulating/recording electrodes in the basolateral amygdala and recording electrode in the contralateral occipital cortex. Rats were divided in three groups (n = 8 for each group) intraperitoneally (i.p.) administered vehicle, TGB 7.3 micromol/kg and TGB 24.3 micromol/kg, respectively, 30 min before stimulation. TGB dose-dependently suppressed the development of the behavioral seizure score and afterdischarge (AD) duration recorded from the amygdala and cortex. Vehicle treated animals displayed at the 16th stimulation an average behavioral score of 4.7 +/- 0.2 (mean +/- SEM) compared to 3.9 +/- 0.2 in the 7.3 micromol/kg TGB treated group and 1.4 +/- 0.3 in the 24.3 micromol/kg TGB treated group. Amygdaloid AD in controls on the 16th stimulation was 92 +/- 10 s compared to 56 +/- 12 s in group 2 and 25 +/- 3 s in group 3. Cortical AD was at the same time 92 +/- 10, 55 +/- 13 and 20 +/- 5 s, respectively. Groups 2 and 3 required four and seven further stimulations, respectively, without TGB administration to reach the AD level in the control group. At the 17th stimulation, rats in group 1 were administered TGB 24.3 micromol/kg and displayed an average behavioral score of 0.5 +/- 0.2. Amygdaloid and cortical AD were both 6 +/- 1 s. Tiagabine 24.3 micromol/kg suppresses both the kindling process and the expression of the fully kindled seizure. PMID- 9223610 TI - Chronic activation of protein kinase C by phorbol ester reduces calcium channel expression in chick sympathetic neurons. AB - Chronic activation of protein kinase C (PKC) has been implicated in regulation of Ca2+ entry responsible for normal development of transmitter properties in cultured sympathetic neurons. The idea that PKC alters the expression of Ca2+ channels was tested using phorbol 12,13-dibutyrate (PDB) which activates PKC and also supports survival of chick sympathetic neurons in the absence of nerve growth factor (NGF). Whole cell voltage-clamp showed that neurons supported by PDB for 2 days had significantly lower Ca2+ current density (0.243 +/- 0.025 pA/microm2) than those supported by NGF (0.356 +/- 0.033 pA/microm2). [125I]omega Conotoxin GVIA binding showed that PDB-supported neurons had significantly lower maximum binding (617 +/- 223 fmol/mg protein) compared with those supported by NGF (1099 +/- 192 fmol/mg protein). These results support the conclusion that chronic activation of PKC limits the expression of N-type Ca2+ channels. A reduction in Ca2+ channel number is consistent with, and could account for the mature type Ca2+ handling and transmitter release properties seen in sympathetic neuro-effector preparations, sympathetic neurons co-cultured with their targets, and neurons supported by PDB. PMID- 9223611 TI - Differential activation of the rat hippocampus and perirhinal cortex by novel visual stimuli and a novel environment. AB - Two groups of rats were shown individual novel visual objects. One group had been familiarised to the environmental context within which the objects were shown, the other experienced the situation for the first time. The activation of neurones in perirhinal cortex and the hippocampal formation was determined by counts of nuclei stained for products of the immediate early gene c-fos. The ratio of counts in the hippocampal formation to that in perirhinal cortex was compared for the two groups: the ratio was significantly higher (4.2:1) in the group experiencing the environment for the first time. Thus, whereas perirhinal neurones are activated by novel rather than familiar objects, hippocampal neurones are preferentially activated by a novel rather than a familiar environment. PMID- 9223612 TI - Human cytomegalovirus infection of breast milk. AB - Human cytomegalovirus is the most common cause of congenital and perinatal infections throughout the world. Primary infection with human cytomegalovirus usually follows a benign course, but the virus remains latent or persistent in the host cell thereafter. Understanding the epidemiology of human cytomegalovirus is a key element in the development of strategies for prevention of infection. Although the actual sites of latency or persistence of human cytomegalovirus infections are still controversial, peripheral blood mononuclear cells and endothelial cells appear to be major sites of infection. Persistent infections caused by human cytomegalovirus could be augmented by a decrease in major histocompatibility complex expression as well as by virus-mediated immune dysfunction. It is possible that specific cellular interactions as well as production of several cytokines are necessary for the reactivation of human cytomegalovirus. Breast-fed infants are susceptible to human cytomegalovirus infection from breast milk. Human cytomegalovirus was isolated more frequently from breast milk at more than 1 month after delivery than from colostrum or early breast milk. Human cytomegalovirus DNA was also not detected in colostrum, but was found in breast milk samples 1 month after delivery. To clarify the role of milk cells and whey in vertical infection by breast feeding, we separated breast milk into milk cells and whey and examined each fraction. Human cytomegalovirus was isolated more frequently from milk whey samples than from cell samples. Human cytomegalovirus particle shedding into whey may be more important in vertical infection by breast milk than cell-to-cell transmission. The supernatant of colostrum did not exert an inhibitory effect on human cytomegalovirus-infected cells. Serum levels of cell free soluble interleukin-2 receptor of mothers with DNA-positive milk at 1 month after delivery were significantly higher than those of mothers with DNA-negative milk. It is likely that levels of factors such as soluble interleukin-2 receptor in serum are related to the reactivation of human cytomegalovirus which occurs locally in the mammary gland of the lactating mother after delivery. This minireview focuses on recent advances in the study of human cytomegalovirus infection of breast milk. PMID- 9223613 TI - A RE-PCR method to distinguish Listeria monocytogenes serovars. AB - Strains (107) of L. monocytogenes were tested with a PCR-restriction enzyme analysis with two new original primers. A segment of 1395 bp containing the entire iap gene in L. monocytogenes was amplified by the PCR technique. The PCR product was cleaved with the restriction enzymes HindIII and RsaI, and the fragments generated were separated by gel electrophoresis. Two groups of serovars were obtained: one group contained serovars 1/2a and 1/2c, the other group contained serovars 1/2b, 3b and 4b. The PCR-restriction enzyme analysis method described in this paper could be a useful tool for the unambiguous division of L. monocytogenes into two serovar groups, and it could be used to study the evolution of different serotypes and groups of serotypes in foods produced in the same processing plant and processed during the same month. The RE-PCR method used can give a rapid confirm at the subgroup level in the laboratory of an epidemiological association between human disease and suspected sources of contaminated food. PMID- 9223614 TI - Augmented inhibition of growth of Candida albicans by neutrophils in the presence of lactoferrin. AB - The combined inhibitory effects of neutrophils and lactoferrins on the growth of Candida albicans were examined. Murine or human neutrophils partially inhibited growth of C. albicans when cultured with C. albicans in vitro. The growth inhibition was augmented by a combination of neutrophils and more than 30 microg/ml of bovine lactoferrin or 1 microg/ml of human lactoferrin, concentrations less than 1/10-1/200 their inhibiting concentrations when used alone. The inhibition of C. albicans was also enhanced by combination of neutrophils and bovine apolactoferrin or iron-bound holo-lactoferrin, but not by transferrin. Combination effects of neutrophils and lactoferrin were also observed in a condition where there was no contact between neutrophils and Candida cells. These results suggest that neutrophils inhibit the growth of C. albicans regardless of whether there is direct contact between them and Candida cells: neutrophil growth inhibition effects were augmented in the presence of a physiological concentration of lactoferrin, perhaps through some action of lactoferrin other than chelation of ferric ion. PMID- 9223615 TI - Molecular analysis of the genes mediating Salmonella invasion. AB - To identify invasion determinants, a genomic library of Salmonella typhimurium was cloned into a cosmid vector, pLA2917. A clone, pSI623 which was invasive for HEp-2 and Henle-407 epithelial cells, was subcloned into a plasmid vector, pGEM 7Z to define the invasion genes. The subclone, pSV6235 containing a 4.5 kbp fragment of the Salmonella genomic region, was highly invasive for HEp-2 and Henle-407 cells, compared with other subclones whose Salmonella genomic regions are 7.4, 6.3, 5.5 and 1.4 kbp, respectively. This study reflects that the invasion efficiency of the host strain, Escherichia coli to HEp-2 and Henle-407 cell lines was significantly increased by the introduction of the genomic region of the virulent S. typhimurium. Restriction enzyme analysis showed that there was a single PstI site on the 27 kbp of Salmonella genomic region of pSI623, and no PstI site was found on the 4.5 kbp of genomic region of pSV6235. This finding suggests that the invasion related genes different from the inv and spa gene clusters which were identified in S. typhimurium, may exist in genomic DNA of S. typhimurium. PMID- 9223616 TI - RAPD analysis of environmental, food and clinical isolates of Campylobacter spp. AB - The typing of Campylobacter is relatively poorly developed compared to that of the Enterobacteriaceae, and new molecular methods may provide useful approaches. The polymerase chain reaction was used to amplify randomly primed genomic DNA from Campylobacter isolates with an optimised randomly amplified polymorphic DNA protocol. Groups of isolates were analysed from chicken house environmental sources, chicken joints from retail sources, patients suffering from clinical disease and laboratory culture collections. Amplicons were separated by agarose gel electrophoresis, stained with ethidium bromide, and banding patterns captured in a digital form for computer analysis with GelCompar software. The method gave 100% typability and reproducibility for the isolates investigated and proved a useful technique for the epidemiological analysis of Campylobacter. Computer based analysis of the randomly amplified polymorphic DNA generated profiles allowed relationships between isolates to be studied at the molecular level resulting in some indication of molecular correlates of the origins of isolates. PMID- 9223617 TI - Inhibition of Salmonella typhimurium adhesion to Caco-2 cell cultures by Lactobacillus strain GG spent culture supernate: only a pH effect? AB - The effect of Lactobacillus GG and its spent culture supernate on the adhesion of Salmonella typhimurium to Caco-2 cells was investigated. Lactobacillus GG cells which had adhered to Caco-2 monolayers prior to the addition of S. typhimurium did not inhibit the adhesion. Adhesion of S. typhimurium was reduced in the presence of spent culture supernate from MRS broth cultures (spent culture supernate I). Similar inhibition was observed with acidified fresh MRS. The viability experiments with Caco-2 cells indicated that the inhibition was presumably due to the death of cells under acidic conditions. Adhesion of S. typhimurium was reduced by pre-treating the bacteria with spent culture supernate I or with acidified MRS and whey broth prior to adhesion to Caco-2 monolayers. Pre-treatment with spent culture supernate II (from whey broth cultures) did not influence the adhesion. No inhibition was detected at neutral pH values. Therefore, the observed inhibition of S. typhimurium adhesion to Caco-2 monolayers with spent culture supernate I was most likely a pH effect. PMID- 9223618 TI - Non-seasonal viral and bacterial episode of diarrhoea in the Jordan Valley, West of Jordan. AB - A non-seasonal diarrhoeal episode in the Jordan Valley occurred over a 2-month period, during which no traditional enteropathogens were detected by the health authority laboratories. A total of 17 diarrhoeal stool specimens from infants, young children and adults were randomly collected and delivered to our laboratories to investigate the presence of unusual aetiological agents. Stools were examined for parasites, ova, viruses and cultured for bacterial pathogens. A multiplex polymerase chain reaction was developed to investigate the involvement of diarrhoeagenic Escherichia coli in this episode. Recognised pathogenic organisms were detected in 8 out of 17 of the diarrhoeatic patients, one patient of whom had a mixed infection with two agents. Rotavirus, enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), and enterotoxigenic E. coli (ETEC) were found to be associated with the diarrhoea. EIEC was the most common enteropathogen detected (4 out of 17) followed by rotavirus (3 out of 17). One of the EIEC isolates detected in one patient was associated with rotavirus. The clinical features of the diarrhoeatic patients were remarkably similar, regardless of aetiology. This study reveals the identity of pathogenic agents that are not detected by traditional methods employed by the health authority laboratories, which emphasise the urgent need for developing the current diagnostic techniques. PMID- 9223619 TI - Annexins and membrane dynamics. PMID- 9223620 TI - The roles of NSF, SNAPs and SNAREs during membrane fusion. PMID- 9223621 TI - Synthesis and decay of calmodulin-ubiquitin conjugates in cell-free extracts of various rabbit tissues. AB - Calmodulin is the natural substrate for ubiquitin-ligation by the enzyme ubiquitin-calmodulin ligase (uCaM-synthetase; EC 6.3.2.21). The activity of this ligase is regulated by the binding of the second messenger Ca2+ to the substrate calmodulin, which increases the activity ca. 10-fold. Up till now, two components of the ligase could be identified: uCaM Syn-F1 and uCaM Syn-F2, the first of which binds to ubiquitin and the second which binds to calmodulin. Since the physiological role of this enzyme is still unclear, this study was designed to examine whether the activity of uCaM-Synthetase in 40,000 x g tissue supernatants correlates with the calmodulin content in the various tissues. In reticulocytes, spleen, erythrocytes, testis and brain, which are rich in uCaM synthetase, the tissue contents calculated on the basis of activity measurements were between 4 80-fold higher than in red and white skeletal muscle. These activities did not correlate with the respective calmodulin contents of the tissues indicating that other factors were determining these enzyme levels. A second aim was to gain information on the role of the ATP-ubiquitin-dependent proteolytic pathway in those tissues displaying uCaM synthetase activity. In the reticulocyte system which contains the classical ATP-ubiquitin-dependent proteolytic pathway as measured with 125I-BSA, no ubiquitin-dependent degradation of calmodulin could be detected. We therefore examined the other tissues of the rabbit with the substrate 125I-BSA and succeeded in finding a ubiquitin-independent ATP-dependent proteolytic activity in every case but no ubiquitin-dependent activity. The ubiquitin-independent activity was highest in smooth muscle and red skeletal muscle being ca. 3-4-fold higher than in lung and testis. In 50% of the tissue crude extracts the time curve of calmodulin ubiquitylation progressed through a maximum indicating a dynamic steady state based on conjugate synthesis and decay. If a ubiquitylation pulse of 30 min was followed in liver crude extracts by the addition of EGTA, which specifically inhibits ubiquityl-calmodulin synthesis, a half-life of calmodulin-conjugate decay of 15-20 min is observed. A similar conjugate half-life of ca. 30 min was observed after addition of EDTA excluding that conjugate decay is due to an ATP-dependent proteolytic process. Studying the decay of purified ubiquitin-125I-BH-calmodulin conjugates in cell-free reticulocyte extracts led to the discovery of an ATP-independent isopeptidase activity which splits ubiquitin-calmodulin conjugates without leading to detectable calmodulin fragments. The rapid decay of ubiquitin-calmodulin conjugates in tissue extracts can therefore be plausibly explained by a ubiquityl calmodulin splitting isopeptidase activity. PMID- 9223622 TI - Role of glucocorticoid on interleukin-6-induced cellular functions in the mouse macrophage cell line (Mm 1). AB - To discover a role of glucocorticoid on interleukin-6 (IL-6)-induced responses of a macrophage, we investigated the effect of IL-6 and/or dexamethasone (Dex) on cellular functions of a mouse macrophage cell line (Mm1 cells). The results obtained were as follows. (1) Dex decreased the accumulation of tumor necrosis factor-alpha induced by IL-6, whereas nitric oxide production was enhanced by Dex. Moreover, the enhancement of nitric oxide production could be demonstrated to be associated with stimulation of iNOS mRNA expression by the Dex treatment. (2) Cytotoxic activity of Mm1 cells on mouse B16 melanoma cells was much more enhanced by the co-treatment of IL-6 with Dex than IL-6 treatment alone. (3) Dex promoted further the suppression of proliferation induced by IL-6. (4) DNA fragmentation, introduced by the treatment of cells with IL-6, was further enhanced in the presence of Dex. PMID- 9223623 TI - Atypical angiotensin II receptors coupled to phosphoinositide turnover/calcium signalling in catfish hepatocytes. AB - In catfish (Ictalurus punctatus) hepatocytes angiotensin II induced an immediate increase in cytosolic Ca2+ concentration. Other angiotensin analogues also induced this effect including: human angiotensin II, fish angiotensin II, human angiotensin III, human angiotensin I, fish angiotensin I and saralasin. CGP 42112A induced a very small effect at the highest concentration tested and angiotensin IV was without effect. Angiotensin II also increased the resynthesis of phosphatidylinositol and the production of IP3. These physiological effects were not blocked by losartan (AT1-selective antagonist) or PD 123177 (AT2 selective antagonist). [125I]Angiotensin II bound to liver plasma membranes in a saturable fashion with high affinity (K(D) 2.7 nM) and a B(max) of 185 fmol/mg of protein. Binding competition experiments showed the following order of potency: human angiotensin II = fish angiotensin II > human angiotensin III > or = human angiotensin I = fish angiotensin I. These sites were insensitive to losartan or PD 123177. The data indicate that the angiotensin II receptors expressed in catfish hepatocytes are coupled to the phosphoinositide turnover/calcium mobilization signal transduction pathway and are atypical receptors, i.e., pharmacologically distinct from mammalian AT1 and AT2 receptors. PMID- 9223624 TI - Regulation of thrombospondin-1 production by angiotensin II in rat heart endothelial cells. AB - Thrombospondin-1 (TSP-1) is synthesized, secreted, and incorporated into the extracellular matrix by a variety of cells, including the endothelial cells. Addition of angiotensin II (AII) significantly induced TSP-1 mRNA in rat heart derived endothelial cells. TSP-1 mRNA levels reached a plateau within 2 h after the addition of AII and decreased after 5 h. The induction was superinduced by cycloheximide and blocked by actinomycin D. Losartan, an AT1 receptor antagonist, could abolish the induction of TSP-1 mRNA by AII. Phorbol 12-myristate 13-acetate (TPA) was found to enhance TSP-1 mRNA level whereas a protein kinase C inhibitor, H7, was shown to block the induction. Immunoblot analysis revealed that TSP-1 was detectable in the medium 4 h after AII stimulation. Our results suggest that the upregulation of TSP-1 by AII represents an important mechanism leading to perivascular fibrosis in the heart. PMID- 9223625 TI - Expression of a bacterial endo (1-4)-beta-glucanase gene in mammalian cells and post translational modification of the gene product. AB - An endo (1-4)-beta-glucanase gene C6.5 from Bacillus subtilis has been expressed in Chinese hamster ovary (CHO) cells and pancreatic 266-6 cells. The fusion gene, stably transfected into CHO cells consisted of the mouse Amy-2.2 signal peptide coding sequence and the endoglucanase gene C6.5 transcribed from the early SV40 promoter/enhancer, using the dihydrofolate reductase gene as a selective marker. The gene construct transfected into pancreatic 266-6 cells consisted of the mouse Amy-2.2 promoter/enhancer and signal peptide coding sequence and the same C6.5 sequences using the xanthine-guanine phosphoribosyl transferase gene (gpt) as the selective marker. The stably transfected CHO cells synthesized endoglucanase at 1.1 U/mg cell protein in a 72 h culture, with 89% of the activity secreted into the culture fluid in a glycosylated form of 66 kDa as compared with the unglycosylated 53 kDa form expressed in E. coli. Glycosylation did not change the specific activity, protease resistance, or cellulose binding of the endoglucanase as compared to the unglycosylated form of the enzyme from E. coli. The level of expression in the stably transfected pancreatic cells was substantially lower at 0.3 mU/mg cell protein with all detectable activity present in the culture fluid. The secreted enzyme from pancreatic cells was glycosylated with a mass similar to that secreted from CHO cells. PMID- 9223627 TI - Regulation of protein turnover by glutamine in heat-shocked skeletal myotubes. AB - Skeletal muscle accounts for approximately one-half of the protein pool in the whole body. Regulation of protein turnover in skeletal muscle is critical to protein homeostasis in the whole body. Glutamine has been suggested to exert an anabolic effect on protein turnover in skeletal muscle. In the present work, we characterized the effect of glutamine on the rates of protein synthesis and degradation in cultured rat skeletal myotubes under both normal and heat-stress conditions. We found that glutamine has a stimulatory effect on the rate of protein synthesis in stressed myotubes (21%, P < 0.05) but not in normal-cultured myotubes. Glutamine shows a differential effect on the rate of degradation of short-lived and long-lived proteins. In both normal-cultured and stressed myotubes, the half-life of short-lived proteins was not altered while the half life of long-lived proteins increased with increasing concentrations of glutamine in a concentration-dependent manner. In normal-cultured myotubes, when glutamine concentration increased from 0 to 15 mM, the half-life of long-lived proteins increased 35% (P < 0.001) while in stressed myotubes, it increased 27% (P < 0.001). We also found that glutamine can significantly (P < 0.001) increase the levels of heat-shock protein 70 (HSP70) in stressed myotubes, indicating that HSP 70 may participate in the mechanism underlying the effect of glutamine on protein turnover. We conclude that in cultured skeletal myotubes the stimulatory effect of glutamine on the rate of protein synthesis is condition-dependent, and that the inhibitory effect of glutamine on the rate of protein degradation occurs only on long-lived proteins. PMID- 9223626 TI - Induction of free radicals in hepatocytes, mitochondria and microsomes of rats by ochratoxin A and its analogs. AB - Oxidative damage may be one of the manifestations of cellular damage in the toxicity of ochratoxin A (OA). OA; its three natural analogs, OB, OC and O alpha; and three synthetic analogs, the ethyl amide of OA (OE-OA), O-methylated OA (OM OA), and the lactone-opened OA (OP-OA) were used to study free radical generation in hepatocytes, mitochondria and microsomes from rats. Electron paramagnetic resonance spectroscopy (EPR) using alpha-(4-pyridyl-1-oxide)-N-tert-butyl nitrone (4-POBN) as a spin trapping agent showed an enhanced free radical generation due to the addition of NADPH to the microsomes. An EPR signal was not observed in the mitochondria and hepatocyte samples when they were treated with a variety of agents. Addition of OM-OA together with NADPH and Fe3+ to the microsomes resulted in a strong EPR signal compared with the other analogs, whereas the signal could be quenched by the addition of catalase. OM-OA does not have a dissociable phenolate group and does not chelate Fe3+. The spin adduct hyperfine splitting constants indicated the presence of alpha-hydroxyethyl radicals resulting from generated hydroxyl radicals, which were trapped by 4-POBN. The results also suggested that the production of hydroxyl radicals by OA does not require a dissociable phenolate group or the prior formation of an OA-Fe complex. PMID- 9223628 TI - Isolation of a Dictyostelium discoideum 14-3-3 homologue. AB - A 1.0 kb cDNA clone (Dd14-3-3) encoding a 14-3-3 homologue was isolated from a Dictyostelium discoideum cDNA library. The putative Dd14-3-3 protein has highest sequence identity to a barley 14-3-3 isoform (74%). Southern blot analysis suggests that only one 14-3-3 gene is present in the Dictyostelium genome. Highest Dd14-3-3 expression is observed in vegetatively growing cells, and expression decreases during multicellular development. In contrast, Dd14-3-3 protein levels detected immunochemically remained constant during Dictyostelium development. Expression of the Dd14-3-3 cDNA in Saccharomyces cerevisiae complemented the lethal disruption of the two yeast genes encoding 14-3-3 proteins (BMH1 and BMH2). This shows that Dd14-3-3 can fulfil the same function(s) as the yeast 14-3-3 proteins. PMID- 9223629 TI - The pituitary adenylate cyclase activating polypeptide (PACAP I) and VIP (PACAP II VIP1) receptors stimulate inositol phosphate synthesis in transfected CHO cells through interaction with different G proteins. AB - The PACAP receptor (PACAP I receptor, selective for PACAP) and the PACAP II VIP1 receptor (recognizing PACAP and VIP with the same high affinity) were stably expressed in Chinese Hamster Ovary (CHO) cells. Cell lines expressing different receptor densities, as measured by binding saturation curves, were selected. Inositol phosphate production was stimulated dose dependently in all the cell lines by PACAP and VIP, and the order of potency of the agonists was identical to that of high affinity receptor occupancy. The stimulatory effect of a saturating peptide concentration was proportional to the total receptor density. At similar receptor densities, however, the PACAP receptor mediated stimulation was higher than the VIP receptor-mediated stimulation. Pretreatment of the cells with pertussis toxin for 8 h had no effect on receptor densities, did not alter the PACAP stimulated inositol phosphate synthesis by the cells expressing the PACAP I receptor but markedly inhibited the response of the cells expressing the PACAP II VIP1 receptor. Thus, the present results indicate that the two G(s)-coupled PACAP I and PACAP II VIP1 receptors may stimulate IP production. The maximal stimulation depended on the number of receptor expressed; the PACAP I and PACAP II VIP1 receptors probably activated the phospholipase C through G proteins of the G(q), and of the G(i)/G(o) families, respectively. PMID- 9223630 TI - Recognition and manipulation of branched DNA structure by junction-resolving enzymes. AB - The junction-resolving enzymes are a class of nucleases that introduce paired cleavages into four-way DNA junctions. They are important in DNA recombination and repair, and are found throughout nature, from eubacteria and their bacteriophages through to higher eukaryotes and their viruses. These enzymes exhibit structure-selective binding to DNA junctions; although cleavage may be more or less sequence-dependent, binding affinity is purely related to the branched structure of the DNA. Binding and cleavage events can be separated for a number of the enzymes by mutagenesis, and mutant proteins that are defective in cleavage while retaining normal junction-selective binding have been isolated. Critical acidic residues have been identified in several resolving enzymes, suggesting a role in the coordination of metal ions that probably deliver the hydrolytic water molecule. The resolving enzymes all bind to junctions in dimeric form, and the subunits introduce independent cleavages within the lifetime of the enzyme-junction complex to ensure resolution of the four-way junction. In addition to recognising the structure of the junction, recent data from four different junction-resolving enzymes indicate that they also manipulate the global structure. In some cases this results in severe distortion of the folded structure of the junction. Understanding the recognition and manipulation of DNA structure by these enzymes is a fascinating challenge in molecular recognition. PMID- 9223631 TI - Quasi-equivalent viruses: a paradigm for protein assemblies. AB - The structure and assembly of icosahedral virus capsids composed of one or more gene products and displaying quasi-equivalent subunit associations are discussed at three levels. The principles of quasi-equivalence and the related geodesic dome formation are first discussed conceptually and the geometric basis for their construction from two-dimensional assembly units is reviewed. The consequences for such an assembly when three-dimensional protein subunits are the associating components are then discussed with the coordinates of cowpea chlorotic mottle virus (CCMV) used to generate hypothetical structures in approximate agreement with the conceptual models presented in the first section. Biophysical, molecular genetic, and atomic structural data for CCMV are then reviewed, related to each other, and incorporated into an assembly model for CCMV that is discussed with respect to the modular, chemical nature of the viral subunit structure. The concepts of quasi-equivalence are then examined in some larger virus structures containing multiple subunit types and auxiliary proteins and the need for additional control points in their assembly are considered. The conclusion suggests that some viral assembly principles are limited paradigms for protein associations occurring in the broader range of cell biology including signal transduction, interaction of transcription factors and protein trafficking. PMID- 9223632 TI - In vivo analyses of upstream promoter sequence elements in the 5 S rRNA gene from Saccharomyces cerevisiae. AB - Upstream promoter elements of the Saccharomyces cerevisiae 5 S rRNA gene have been characterized by genomic DNase I "footprinting" and by in vivo mutational analyses using base substitutions and deletions. A high copy shuttle-vector was used to efficiently express the mutant 5 S rRNA genes in vivo and a structural mutation in the 5 S rRNA, which was previously shown to be functionally neutral but easily detected by gel electrophoresis, allowed for an accurate measure of gene expression. The results provide direct evidence for upstream regulatory elements which confirms a start site element (sse) from -1 to -8 and identifies a new independent upstream promoter element (upe) centered from about -17 to -20. In contrast to previous reports with reconstituted systems, both elements dramatically affect the efficiency of gene expression and suggest that the saturated conditions which are used in reconstituted studies mask sequence dependence; a dependency that could be physiologically significant and play a role in the regulation of 5 S rRNA expression. The footprint analyses support an extended region of protein interaction as recently observed in reconstituted systems but again provide evidence of significant structural rearrangements when the upstream sequence is changed. PMID- 9223633 TI - Plasmid pIP501 encoded transcriptional repressor CopR binds asymmetrically at two consecutive major grooves of the DNA. AB - Replication of the streptococcal plasmid pIP501 is regulated by the CopR protein and an antisense-RNA (RNAIII). CopR acts as transcriptional repressor at the essential repR promoter pII by binding to inverted repeat IR1 upstream of pII. To further characterize the interaction of CopR with its target, footprinting studies were performed. Methylation interference identified three guanine bases (G240, G242 and G251) in the top strand and two (G252 and G254) in the bottom strand contacted by CopR in the major groove of the DNA. Missing base interference revealed the contribution of the bases in the neighbourhood of these guanine bases to the specific DNA-protein contacts. Phosphate residues essential for CopR binding were determined by ethylation interference. The recognition sequence was localized at the centre of inverted repeat IR1. CopR contacts two consecutive major grooves (site I and II) on the same face of the DNA. Although the two sites share a common sequence motif, neighbouring bases are contacted differently. DNA fragments carrying single mutations in site I or II were analysed by band shift assays. Gel filtration and native gel electrophoresis demonstrated that CopR exists only as a dimer. A sigmoidal binding curve of CopR to its DNA target was observed and allowed the determination of the apparent dissociation constant K(D). The significance of the relatively high apparent K(D) for the role of CopR in pIP501 copy number regulation is discussed. PMID- 9223634 TI - Modified phage peptide libraries as a tool to study specificity of phosphorylation and recognition of tyrosine containing peptides. AB - Tyrosine phosphorylation and protein recognition, mediated by phosphotyrosine containing peptides, play an important role in determining the specific response of a cell, when stimulated by external signals. We have used peptide repertoires displayed by filamentous phage as a tool to study the substrate specificity of the protein tyrosine kinase (PTK) p55(fyn) (Fyn). Peptide libraries were incubated for a short time in the presence of Fyn and phages displaying efficiently phosphorylated peptides were selected by panning over anti phosphotyrosine antibodies. The characterization of the peptides enriched after three phosphorylation/selection rounds allowed us to define a canonical substrate sequence for the kinase Fyn, E-(phi/T)YGx phi, where phi represents any hydrophobic residue. A peptide conforming to this sequence is a better substrate than a second peptide designed to be in accord with the consensus sequence recognised by the Fyn SH2 domain. When the library phosphorylation reaction is carried out in saturation conditions, practically all the tyrosine containing peptides are phosphorylated, irrespective of their context. These "fully modified" peptide libraries are a valuable tool to study the specificity of phosphotyrosine mediated protein recognition. We have used this new tool to identify a family of peptides that bind the PTB domain of the adapter protein Shc. Comparison of the peptide sequences permits us to confirm the essential role of N at position -3, while P often found at position -2 in natural targets is not absolutely required. Furthermore, our approach permits us to reveal an "extended" consensus indicating that residues that do not seem to influence binding in natural peptides can make productive contacts, at least in linear peptides. PMID- 9223635 TI - Epitope mapping by negative selection of randomized antigen libraries displayed on filamentous phage. AB - Since most antibodies directed against protein antigens recognize epitopes composed of several discontinuous segments of the polypeptide chain, attempts to delineate the amino acids constituting these epitopes with the use of linear peptides have generally been unsuccessful. Here, a method is described based on error-prone PCR, phage display and negative selection, whereby amino acid residues constituting the functional epitope are identified in the context of the native protein. First a library of randomized antigen variants containing most single, double and triple amino acid mutants generated by single nucleotide substitutions is produced by error-prone PCR amplification of the DNA sequence encoding the protein antigen. The phage-displayed library is then negatively selected for epitope loss mutants by passing through an affinity matrix derivatized with a specific antibody and positively selected for retention of function. This method was applied to the mapping of the epitopes of two murine monoclonal antibodies (MA-7H11 and MA-3G10) on staphylokinase, a 136 amino acid plasminogen activator secreted by some strains of Staphylococcus aureus. After two negative/positive selection cycles, DNA sequencing of several clones revealed preferential amino acid mutations at positions 35 and 130 (with MA-7H11), and at positions 62, 66 and 136 (with MA-3G10). Affinity measurements of staphylokinase variants carrying single amino acid mutations at these positions confirmed their contribution to the free energy of binding to MA-7H11 and MA-3G10. This approach may be useful for isolating mutants with altered antigenic or functional properties and in general to map critical regions for protein-protein interactions. PMID- 9223636 TI - In vitro binding of the response regulator CitB and of its carboxy-terminal domain to A + T-rich DNA target sequences in the control region of the divergent citC and citS operons of Klebsiella pneumoniae. AB - The genes specifically required for citrate fermentation in Klebsiella pneumoniae form a cluster on the chromosome consisting of two divergently transcribed groups, citCDEFG and citS-oadGAB-citAB. Northern blot analyses described here and elsewhere indicate that each group forms an operon. The transcriptional start sites of citC and citS, which were mapped in this work by primer extension, are separated by a stretch of 193 bp with an extraordinary high A + T content of 67%. Expression of the citrate fermentation genes was recently shown to be positively controlled by a two-component signal transduction system encoded by the promoter distal genes of the citS operon, citA (sensor kinase) and citB (response regulator). As a first step towards the functional characterization of CitB, we analysed its DNA-binding properties. To this end, the entire CitB, its N-terminal receiver domain (CitBN), and its C-terminal output domain (CitBC), all modified by a (His)6-tag, were purified. CitB(His) and CitBN(His) could be phosphorylated either with acetylphosphate or with ATP plus MalE-CitAC. The latter protein contains the kinase domain of CitA fused to the C terminus of the maltose-binding protein. Upon phosphorylation, CitB(His) became more resistant towards limited proteolysis by trypsin, reflecting substantial changes in tertiary structure. In gel retardation assays, CitB(His) bound specifically to the citC-citS intergenic region. The retardation pattern changed significantly upon phosphorylation and the apparent binding affinity increased 10 to 100-fold. Depending on the protein concentration, four different phospho-CitB(His)-DNA complexes could be resolved, suggesting the presence of multiple binding sites between citC and citS. DNase I footprints revealed two protected regions extending maximally from -55 to -89 relative to the citS transcription start and from -50 to -96 relative to the citC transcription start. Gel retardation and DNase I footprint assays with CitBC(His) showed that the C-terminal domain is sufficient for specific DNA binding. Since its properties were similar to that of unphosphorylated CitB(His), an essential role of the N-terminal receiver domain in high-affinity DNA binding was indicated. The positions of the binding sites for CitB and of putative recognition sequences for the cAMP receptor protein suggested a model for the interaction of these activators with RNA polymerase. PMID- 9223637 TI - In vivo determination of RNA structure-function relationships: analysis of the 790 loop in ribosomal RNA. AB - The 790 loop is a conserved hairpin located between positions 786 and 796 of Escherichia coli 16 S rRNA that is required for ribosome function. Using a novel genetic approach, all positions in the loop were simultaneously mutated and functional mutant sequences were selected in vivo. This "instant evolution" experiment revealed that approximately 190 of the 262,144 possible mutant sequences were functional. Analysis of functional mutant sequences allowed discrimination between nucleotides directly involved in protein synthesis and those involved primarily in loop structure. Among the functional mutant sequences, positions 789 and 791 were invariant and extensive covariation was observed among the nucleotides at the base of the loop at positions 787, 788, 794 and 795. NMR and thermodynamic analyses of model 790 hairpins in vitro revealed weak pairing interactions between positions 787 and 795 and between positions 788 and 794 consistent with the in vivo mutational analysis. Functional analysis of site-directed mutants containing all possible nucleotide combinations at positions 787 and 795 in vivo showed that stable base-pairs at these positions prevent subunit association. PMID- 9223638 TI - Molecular characterization of the hdm2-p53 interaction. AB - A number of viral oncogenes target the tumour suppressor protein p53 and inactivate its function. This is an important step in tumourogenesis. The cellular oncogene hdm2 acts through a similar mechanism. It binds the N terminus of p53, thereby interfering with the ability of p53 transcriptionally to activate genes responsible for growth arrest or apoptosis after genotoxic insults. The disruption of the interaction of the two proteins therefore comprises a promising therapeutic target for treatment of the subset of human cancers in which this pathway is active. In this paper we attempt to characterize the p53-hdm2 interaction biochemically. We analyse the potential of a series of peptide inhibitors, derived from previously described mdm2 binding peptide display phage, to disrupt this interaction in ELISA assays. We conclude that F19, W23 and L26 of p53 are critical contact points for p53 binding to hdm2. Furthermore, we show the potential of the monoclonal antibody 3G5 to interfere with binding of p53 to hdm2 in ELISA assays. Consequently, we define the binding site of 3G5 on hdm2 using overlapping peptides derived from the N terminus of hdm2 and phage display libraries. The result indicates L66, Y67 and E69 on hdm2 as critical binding points for 3G5. In electrophoretic mobility shift assay we demonstrate the formation of hdm2-p53 complexes that can be disrupted in the presence of 3G5 or inhibitory peptides. Finally, we describe the effects of NEM and DTT on the interaction between the two molecules in ELISA assays. All our results are discussed in the light of the recently published crystal structure of the mdm2 p53 complex. A striking correspondence between our findings and the crystal structure is revealed. PMID- 9223639 TI - The power stroke of the DnaK/DnaJ/GrpE molecular chaperone system. AB - The molecular chaperone DnaK, the Hsp70 homolog of Escherichia coli, acts in concert with the co-chaperones DnaJ and GrpE. The aim of this study was to identify the particular phase of the peptide binding-release cycle of the DnaK/DnaJ/GrpE system that is directly responsible for the chaperone effects. By real-time kinetic measurements of changes in the intrinsic fluorescence of DnaK and in the fluorescence of dansyl-labeled peptide ligands, the rates of the following steps in the chaperone cycle were determined: (1) binding of target peptide to fast-binding-and-releasing, low-affinity DnaK ATP; (2) DnaJ-triggered conversion of peptide x DnaK x ATP (T state) to slowly-acting, high-affinity peptide x DnaK x ADP x P(i) (R state); (3) switch from R to T state induced by GrpE-facilitated ADP/ATP exchange; (4) release of peptide. Under conditions approximating those in the cell, the apparent rate constants for the T --> R and R --> T conversion were 0.04 s(-1) and 1.0 s, respectively. The clearly rate limiting T --> R conversion renders the R state a minor form of DnaK that cannot account for the chaperone effects. Because DnaK in the absence of the co chaperones is chaperone-ineffective, the T state has also to be excluded. Apparently, the slow, ATP-driven conformational change T --> R is the key step in the DnaK/DnaJ/GrpE chaperone cycle underlying the chaperone effects such as the prevention of protein aggregation, disentangling of polypeptide chains and, in the case of eukaryotic Hsp70 homologs, protein translocation through membranes or uncoating of clathrin-coated vesicles. PMID- 9223640 TI - Structure of ribgrass mosaic virus at 2.9 A resolution: evolution and taxonomy of tobamoviruses. AB - Ribgrass mosaic virus (RMV) is a member of the tobamovirus group of plant viruses. The structure has been determined at 2.9 A resolution by fiber diffraction methods, and refined by molecular dynamics methods to an R-factor of 0.095. The carboxyl-carboxylate interactions that drive disassembly in tobamoviruses are present in RMV, but are very different from those in other tobamoviruses. RMV has some of the structural features of a subgroup I tobamovirus, a smaller number from subgroup II, and a number that appear to be unique to the RMV cluster of viruses. The structural studies confirm the evolutionary and taxonomic separation of the RMV cluster from both subgroup I and subgroup II tobamoviruses. PMID- 9223641 TI - Molecular recognition in the HIV-1 capsid/cyclophilin A complex. AB - The HIV-1 capsid protein (CA) makes an essential interaction with the human peptidyl prolyl isomerase, cyclophilin A (CypA), that results in packaging of CypA into the virion at a CA to CypA stoichiometry of approximately 10:1. The 231 amino acid residue capsid protein is composed of an amino-terminal CypA binding domain (1 to approximately 151; CA151) and a carboxyl-terminal dimerization domain (approximately 151 to 231). We find that CypA binds dimeric CA and monomeric CA151 with identical intrinsic affinities (K[d] = 16(+/-4) microM). This result demonstrates that capsid dimerization and cyclophilin A binding are not thermodynamically coupled and suggests that the substoichiometric ratio of CypA in the HIV-1 virion results from the intrinsic stability of the CA/CypA complex. In the known co-crystal structure of the CA151/CypA complex, CypA binding is mediated exclusively by an exposed capsid loop that spans residues Pro85 to Pro93. The energetic contributions to CypA binding were quantified for each residue in this loop, and the results demonstrate that the Gly89-Pro90 dipeptide is the primary cyclophilin A recognition motif, with Pro85, Val86, His87, Ala88, and Pro93 also making energetically favorable contacts. These studies reveal that the active site of CypA, which can catalyze the isomerization of proline residues in vitro, also functions as a sequence-specific, protein binding motif in HIV-1 replication. PMID- 9223642 TI - The structures and relative stabilities of d(G x G) reverse Hoogsteen, d(G x T) reverse wobble, and d(G x C) reverse Watson-Crick base-pairs in DNA crystals. AB - We have solved the structures of the homoduplex d(Gm5CGCGCG)2, and the heteroduplexes d(GCGCGCG)/d(TCGCGCG) and d(GCGCGCG)/d(CCGCGCG). The structures form six base-pairs of identical Z-DNA duplexes with single nucleotides overhanging at the 5'-ends. The overhanging nucleotide from one strand remains stacked and sandwiched between the blunt-ends of two adjacent Z-DNA duplexes, while the overhanging base of the opposing strand is extra-helical. The stacked and the extra-helical bases from adjacent duplexes pair to form a distorted d(G x G) reverse Hoogsteen base-pair in the d(Gm5CGCGCG)2 homoduplex, and d(G x T) reverse wobble and d(G x C) reverse Watson-Crick base-pairs in the d(GCGCGCG)/d(TCGCGCG) and d(GCGCGCG)/d(CCGCGCG) heteroduplexes, respectively. Interestingly, only the d(G,T) and d(G x C) base-pairs were observed in the heteroduplexes, suggesting that both the d(G x T) reverse wobble and d(G x C) reverse Watson-Crick base-pairs are more stable in this crystal environment than the d(G x G) reverse Hoogsteen base-pair. To estimate the relative stability of the three types of reverse base-pairs, crystals were grown using various mixtures of sequences and their strand compositions analyzed by mass spectrometry. The d(G x C) reverse Watson-Crick base-pair was estimated to be more stable by approximately 1.5 kcal/mol and the d(G x T) reverse wobble base-pair more stable by approximately 0.5 kcal/mol than the d(G x G) reverse Hoogsteen base-pair. The step during crystallization responsible for discriminating between the strands in the crystal is highly cooperative, suggesting that it occurs during the initial nucleating event of crystal growth. PMID- 9223643 TI - Structure of a DNA analog of the primer for HIV-1 RT second strand synthesis. AB - The non-self-complementary DNA decamer C-A-A-A-G-A-A-A-A-G/C-T-T-T-T-C-T-T-T-G is a DNA/DNA analogue of a portion of the polypurine tract or PPT, which is a RNA/DNA hybrid that serves as a primer for synthesis of the (+) DNA strand by HIV reverse transcriptase (RT), and which is not digested by the RNase H domain of reverse transcriptase following (-) strand synthesis. The same unusual conformation that eludes RNase H, thought to be a change in width of minor groove, may also be responsible for the inhibition of HIV RT by minor groove binding drugs such as distamycin and their bis-linked derivatives. The present X ray crystal structure of this DNA decamer exhibits the usual properties of A tract B-DNA under biologically relevant conditions: large propeller twist of base pairs, narrowed minor groove, and a straight helix axis. Groove narrowing is fully developed in the A-A-A-A region, but not in the A-A-A region, which previous investigators have proposed as being too short to exhibit typical A tract properties. The RNA/DNA hybrid produced by HIV reverse transcriptase during (-) strand synthesis presumably forms a "heteromerous" or H-helix with narrower minor groove than an A-helical RNA/RNA duplex. If the narrowing of minor groove in A-tract H-helices is comparable to that seen in A-tract B-helices, then the narrowed minor groove of the polypurine tract could make the second primer site both (1) impervious to RNase H digestion, and (2) susceptible to inhibition by minor groove binding drugs. PMID- 9223644 TI - Sequence-dependent crossed helix packing in the crystal structure of a B-DNA decamer yields a detailed model for the Holliday junction. AB - The structure of the B-DNA decamer d(CGCAATTGCG)2 has been determined by X-ray diffraction analysis to a resolution of 2.3 A and an R-factor of 17.7%. The decamer crystallises in the monoclinic space group C2 and packs with a crossed arrangement of helices and a unique crossing contact distinct from all other decamer structures. This is believed to be a direct result of the sequence dependent minor groove width of the duplex. Crossed helix structures of DNA are valuable starting points for modelling studies of the Holliday junction. Two unique sites are observed at the cross-over junction where strand exchange may occur. A Holliday junction model has been constructed for each case and modelled using molecular mechanics and dynamics techniques. One of these models was found to be fully consistent with the available physical data. PMID- 9223645 TI - The cyclic AMP receptor promoter DNA complex: a comparison of crystal and solution structure by quantitative molecular electrooptics. AB - The complexes formed between the cyclic AMP receptor and three different promoter DNA fragments, including a synthetic 30 bp fragment with the sequence used for determination of the crystal structure, have been analysed in solution by measurements of the electric dichroism (ED) at an ionic strength of 105 mM, using a special instrument based on cable discharge. The ED of the protein is negligible and, thus, the ED of the complexes is determined by the DNA and its orientation relative to the protein. The complex formed between the cyclic AMP receptor with the 30 bp fragment is characterized by a positive ED, indicating that the electric dipole is perpendicular relative to the direction of the helix; moreover, the dipole changes its nature from an induced one for the free DNA to a permanent one of 3.0 x 10(-27) Cm for the complex; both the limiting value of the ED +0.3 and the dichroism decay time constant of 62 ns found for the complex (free DNA: 52 ns; 20 degrees C) demonstrate bending of the DNA double helix. All these parameters are calculated quantitatively from the crystal structure: bead model simulations are used to derive the coefficients of rotational diffusion and to define the center of diffusion, which is the reference for calculation of the dipole vector; the dipole vector is then the basis for calculation of the limit value of the dichroism; the time constants are derived from the diffusion coefficients of the bead model The calculated parameters are in very satisfactory agreement with the experimental ones, demonstrating agreement of the structures in the crystal and in solution with respect to their essential features. These results also demonstrate the utility of electrooptical procedures for a quantitative comparison of crystal or model structures with structures in solution. While the crystal structure has been determined only for the complex with the 30 bp promoter fragment, it has been relatively simple to extend measurements of the ED to complexes formed with a 40 bp and a 203 bp promoter fragment: the data obtained for a 40 bp fragment with the consensus binding sequence are quite similar to those obtained for the 30 bp promoter, whereas the data obtained for the 203 bp promoter clearly show a much higher degree of protein induced bending with a bending angle of approximately 180 degrees. PMID- 9223646 TI - Crystal structures and mechanism of 6-phospho-beta-galactosidase from Lactococcus lactis. AB - The initial structural model of 6-phospho-beta-galactosidase from Lactococcus lactis was refined to an R-factor of 16.4% (R[free] = 23.6%) to 2.3 A resolution (1 A = 0.1 nm), and the structures of three other crystal forms were solved by molecular replacement. The four structural models are essentially identical. The catalytic center of the enzyme is approximately at the mass center of the molecule and can only be reached through a 20 A long channel, which is observed with an "open" or "closed" entrance. The closed entrance is probably too small for the educt lactose-6-phosphate to enter, but large enough for the first product glucose to leave. Among the presented structures is a complex between an almost inactive mutant and the second product galactose-6-phosphate, which is exclusively bound at side-chains. A superposition (onto the native enzyme) of galactose-6-phosphate as bound to the mutant suggests the geometry of a postulated covalent intermediate. The binding mode of the educt was modeled, starting from the bound galactose-6-phosphate. A tightly fixed tryptophan is used as a chopping-board for splitting the disaccharide, and several other aromatic residues in the active center cavity are likely to participate in substrate transport/binding. PMID- 9223647 TI - Crystal structure of an oligomer of proteolytic zymogens: detailed conformational analysis of the bovine ternary complex and implications for their activation. AB - The pancreas of ruminants secretes a 100 kDa non-covalent ternary complex of the zymogen of a metalloexopeptidase, carboxypeptidase A, and the proforms of two serine endopeptidases, chymotrypsin C and proteinase E. The crystal structure of the bovine complex has been solved and refined to an R-factor of 0.192 using synchrotron radiation X-ray data to 2.35 A resolution. In this heterotrimeric complex, the 403 residue procarboxypeptidase A takes a central position, with chymotrypsinogen C and proproteinase E attached to different surface sites of it. The procarboxypeptidase A subunit is composed of the active enzyme part and the 94 residue prodomain, similar to the monomeric porcine homologous form. The 251 residue subunit chymotrypsinogen structure, the first solved of an anionic (acidic pI) chymotrypsinogen, exhibits characteristics of both chymotrypsinogen A and elastases, with a potential specificity pocket of intermediate size (to accommodate apolar medium-sized residues) although not properly folded, as in bovine chymotrypsinogen A; this pocket displays a "zymogen triad" characteristic for zymogens of the chymotrypsinogen family, consisting of three non-catalytic residues (one serine, one histidine, and one aspartate) arranged in a fashion similar to the catalytic residues in the active enzymes. Following the traits of this family, the N terminus is clamped to the main molecular body by a disulphide bond, but the close six residue activation segment is completely disordered. The third zymogen, the 253 residue proproteinase E, bears close conformational resemblance to active porcine pancreatic elastase; its specificity pocket is buried, displaying the second "zymogen triad". Its five N-terminal residues are disordered, although the close activation site is fixed to the molecular surface. The structure of this native zymogen displays large conformational differences when compared with the recently solved crystal structure of bovine subunit III, an N-terminally truncated, non-activatable, proproteinase E variant lacking the first 13 residues of the native proenzyme. Most of the prosegment of procarboxypeptidase A and its activation sites are buried in the centre of the oligomer, whilst the activation sites of chymotrypsinogen C and proproteinase E are surface-located and not involved in intra or inter-trimer contacts. This organization confers a functional role to the oligomeric structure, establishing a sequential proteolytic activation for the different zymogens of the complex. The large surface and number of residues involved in the contacts among subunits, as well as the variety of non-bonded interactions, account for the high stability of the native ternary complex. PMID- 9223648 TI - The 1.8 A crystal structure of winged bean albumin 1, the major albumin from Psophocarpus tetragonolobus (L.) DC. AB - Winged bean albumin-1 (WBA) is the main seed albumin of Psophocarpus tetragonolobus, a legume that has excellent potential as a protein-rich food source for humid tropical climates. WBA crystallises in a tetragonal space group and the structure was solved by X-ray crystallography with a combination of multiple isomorphous replacement using four heavy atom derivatives and molecular replacement with a model based on the structure of Erythrina caffra trypsin inhibitor (ETI). Refinement of the structure proceeded to 1.8 A. WBA has a beta trefoil fold, similar to that found in the STI-Kunitz type trypsin inhibitors. The final structure has an overall R-factor of 19% for 15 to 1.8 A resolution data, all residues in the allowed regions of the Ramachandran plot, and good agreement with ideal geometry. WBA has sequence similarity with the STI-Kunitz trypsin inhibitors, including the apparent conservation of the functional reactive site residue, lysine 64, at the position of the scissile bond (position P1) in the STI-Kunitz type trypsin inhibitors, however, WBA does not inhibit trypsin. The reason for the lack of inhibitory activity against trypsin is clearly evident from the structure. The loop corresponding to the inhibitory loop in the STI-Kunitz trypsin inhibitors does not conform to the canonical conformation of the inhibitory loops of the "small inhibitors". The lysine residue assigned to the P1 position from sequence alignments is instead part of a four amino acid insertion between residues structurally equivalent to residues P1 and P2 of the inhibitors. PMID- 9223649 TI - Importance of electrostatic interactions in the rapid binding of polypeptides to GroEL. AB - The question of how chaperones rapidly bind non-native proteins of very different sequence and function has been examined by determining the effect of ionic strength on the refolding of barnase on GroEL, and on the thermal denaturation of barnase in the presence of GroEL and SecB. Both chaperones bind the denatured state of barnase, so lowering the T(m) value. The refolding of barnase in the presence of GroEL is multiphasic, the slowest phase corresponding to the refolding of a singly bound molecule of barnase in the complex with GroEL. The fastest phase is related to the association of barnase and GroEL. At high ratios of GroEL to barnase and low ionic strength (less than 200 mM) this fast phase corresponds to the observed rate of binding. The rate of association of barnase and GroEL was found to be highly dependent on ionic strength, and at high ionic strength (greater than 600 mM) the majority of barnase molecules escaped binding and refolded free in solution. The data are consistent with an initial, transient, ionic interaction between barnase and GroEL, before hydrophobic binding occurs, allowing diffusion-controlled association and slow dissociation of unfolded polypeptide. PMID- 9223650 TI - Multiple sequence threading: an analysis of alignment quality and stability. AB - Methods that compare a protein sequence directly to a structure can be divided into those that construct a molecular model (threading methods) and those that perform a sequence alignment with the structure encoded as a sequence of structural states (one-dimensional/three-dimensional (1D/3D) matching). The former take into account the internal packing of the molecule but the latter do not. On the other hand, it is simple to include multiple sequence data in a 1D/3D comparison but difficult in a threading method. Here, a protein sequence/structure alignment method is described that uses a combination of matching predicted and observed residue exposure, predicted and observed secondary structure (1D/3D) together with pairwise packing interactions in the core (threading). Using a variety of distantly related and analogous protein structures, the multiple sequence threading (MST) method was compared to a single sequence threading (SST) method (that uses complex potentials of mean-force) and also to a multiple sequence alignment (MSA) program. It was found that the MST method produced alignments that were better than the best that could be obtained with either the SST or MSA method. The method was found to be stable to error in both secondary structure prediction and predicted exposure and also under variation of the key parameters (fully described in an Appendix). The contribution of the pairwise term was found to be small but without it, the correct alignments were less stable and structurally unreasonable deletions were observed when matching against larger structures. Using the parameters derived for alignment, the method was able to recognise related folds in the structure databank with a specificity comparable to other methods. PMID- 9223651 TI - Effects of tepoxalin, a dual inhibitor of cyclooxygenase/5-lipoxygenase, on events associated with NSAID-induced gastrointestinal inflammation. AB - Prostaglandins and thromboxanes are products of arachidonic acid metabolism via the cyclooxygenase (CO) enzyme and are responsible for the pain and swelling common to sites of inflammation. Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit the production of these substances and are used in the treatment of inflammatory diseases such as arthritis. However, one of the major side-effects of NSAID therapy is gastric ulceration. It is possible that inhibition of prostaglandin production and a related increase in the formation of leukotrienes via the 5-lipoxygenase (5-LO) enzymatic pathway are responsible for attracting inflammatory cells, causing local sites of inflammation and producing ulceration. To determine the effects of 5-LO inhibition on this hypothesis, studies were performed in rats to evaluate the effects of tepoxalin, a dual CO/LO inhibitor on leukotriene B4 levels in gastric mucosa and neutrophil adhesion in mesenteric venules. In rats, chronic oral administration of an NSAID, indomethacin (2 mg/kg daily over 4 days), resulted in 40% mortality, accompanied by intestinal adhesions and perforations when evaluated 24 h after the fourth dose of drug. Additionally, neutrophil adhesion was increased in the mesenteric venules and cell infiltration was evident in the mesenteric interstitium. These gastrointestinal side-effects were inhibited in a separate group of rats administered tepoxalin (20 mg/kg, p.o) 30 min prior to each daily indomethacin treatment. Further studies were performed to determine tepoxalin's effects on early events associated with NSAID-induced gastrointestinal inflammation, including neutrophil adhesion, lipid peroxide generation and LTB4 production. Indomethacin (100 mg/kg, p.o.) produced elevated levels of LTB4 in rat gastric mucosa 90 min after administration. Additionally, neutrophil adhesion in mesenteric venules was increased at this dose and with the administration of another NSAID, naproxen. No generation of lipid peroxides was evident in the gastric mucosa at this timepoint. Tepoxalin (up to 400 mg/kg, p.o.) did not have an effects on gastric mucosal LTB4 generation and lipid peroxide levels. A decrease in neutrophil adhesion was observed at the highest dose. In another study, pretreatment with tepoxalin (ED50=7.5 mg/kg, p.o.) or the selective 5-LO inhibitor zileuton (100 mg/kg, p.o.) prevented the increases in gastric mucosal LTB4 levels and neutrophil adhesion induced by indomethacin (100 mg/kg, p.o.). These data suggest that LO inhibition may play a vital role in the prevention of NSAID-induced gastric inflammation, providing insight into the lack of ulcerogenicity with tepoxalin and new approaches to anti-inflammatory therapy which may prevent gastric side effects. PMID- 9223652 TI - Eicosanoid production by placental and amnion tissues from control and non insulin-dependent diabetic rats. Influence of oxytocin in the incubating medium. AB - Eicosanoid production by intrauterine tissues from control and neonatal streptozotocin induced diabetic rats during late pregnancy was evaluated. In diabetic placenta the release of 6-keto-PGF1alpha was found diminished when compared to controls. In addition, LTB4 generation was increased in diabetic placenta. No alterations in the production of TXA2, PGE2, PGE1 and PGF2alpha was found when diabetic and control placenta were compared. In amnion tissue a decreased generation of 6-keto-PGF1alpha was observed in the diabetic group, but no alteration in any other eicosanoid evaluated was found. Oxytocin (5 mU/ml, in vitro), which increases prostaglandin synthesis in rabbit and human amnion tissues, did not modify eicosanoid generation in control rat amnion. In contrast, in diabetic amnion the presence of oxytocin further decreased the release of 6 keto-PGF1alpha and diminished PGE1 generation. The present results suggest that this mildly diabetic state induces alterations in eicosanoid production in intrauterine tissues, abnormalities probably enhanced during parturition, when endogenous concentrations of oxytocin are elevated. PMID- 9223653 TI - Pharmacophore requirements in new series of pyridazinyl alkanoic acids, N [(pyridazin-2-yl) alkyl] succinyl and glutaryl amides, inhibitors of thromboxane A2 biosynthesis. AB - New series of 5-benzyl-6-methyl-4-oxo pyridazin-2-yl alkanoic acids, N [(pyridazin-2-yl)alkyl] succinyl and glutaryl amides have been synthesized and evaluated in vitro as TXA2 biosynthesis inhibitors. The experiments were carried out using arachidonic acid (32.8 microM) as a substrate and horse platelet microsomes as sources of TXA2 synthase. The presence of TXB2, a stable metabolite of TXA2, was determined by RIA. The potency of active compounds (1.10(-4) < IC 50 < 1.10(-6) M) greatly depends on the length of the chain at the N-2 position on the pyridazine ring. Furthermore, enzyme inhibition in vitro is increased with the presence of a halogen atom on the aromatic moiety of the benzyl group at C-5. Compound 4f having a pentanoic side chain and a 4-fluoro benzyl moiety was the most active derivative with an IC50 value of 6.69 x 10(-6) M. Molecular modelling studies were done on all the synthesized pyridazinones and on prostaglandin H2 (PGH2) suggesting spatial features and volumes of TXA2 synthase pharmacophore mode in these series of derivatives. PMID- 9223654 TI - Delta-6-desaturase and delta-5-desaturase in human Hep G2 cells are both fatty acid interconversion rate limiting and are upregulated under essential fatty acid deficient conditions. AB - Essential fatty acids are interconverted by desaturases and elongases to eicosanoid precursors. In essential fatty acid deficiency (EFAD) an increased hepatic interconversion of linoleic acid (18:2) to arachidonic acid (20:4n-6) has been demonstrated in vivo. We now cultured Hep G2 cells under EFAD conditions. 20:3n-6 appeared in EFAD cells, but also in controls. After adding 14C-18:2 to the medium, interconversion products and their distribution in different lipids were studied by HPLC. When trace amounts 18:2 were incubated, 38% were converted by the EFAD cells after 21 h, vs 6% by controls. 20% was converted to 20:4 by EFAD cells vs 14% by controls. EFAD cells preferentially distributed more 18:2 and conversion products to neutral fats and to phosphatidyl ethanolamine, but less to cardiolipin than controls did, when incubated with trace amount 18:2, but not with 1 mM 18:2. A relative accumulation of radioactivity in 20:3 was observed. In conclusion; in EFAD Hep G2 cells delta-6- and delta-5-desaturase both were found to be upregulated and eicosanoid precursors were distributed more into phosphatidyl ethanolamine. Delta-5-desaturase had a rate limiting property as well as delta-6-desaturase. PMID- 9223655 TI - Generation second messengers by prostanoids in the iris-sphincter and ciliary muscles of cows, cats and humans. AB - We have examined the generation of second messengers after stimulation of feline, bovine, human iris-sphincter and ciliary muscles by selected prostaglandins (PGs). The tissues, labeled or unlabeled with 3H-myo-inositol, were stimulated by a range of concentrations of 16,16-dimethyl PGE2, 11-deoxy PGE1, 17-phenyl trinor PGE2 and PGF2alpha. In both tissues of all three species, 16,16-dimethyl PGE2 and 11-deoxy PGE1 stimulated the formation of cyclic AMP. Butaprost, an EP2 receptor agonist, which was tested only in feline ciliary muscle, generated cyclic AMP. In the feline iris-sphincter and in bovine and feline ciliary muscles, 17-phenyl trinor PGE2, an EP1 receptor agonist, significantly increased inositol phosphate turnover. The FP receptor agonist, PGF2alpha stimulated inositol phosphate turnover in the bovine, feline, and human iris-sphincter muscles and in human ciliary muscles. Feline and bovine ciliary muscles did not respond to PGF2alpha. These results suggest that EP1 receptors are present in feline iris-sphincter muscle and in bovine and feline ciliary muscles. The EP2 receptors exist in both tissue. These results also suggest the presence FP receptors in bovine, feline, and human iris-sphincter and in human ciliary muscles. Bovine and feline ciliary muscles do not appear to express FP receptors. PMID- 9223656 TI - Prostaglandin levels in BL6 melanoma cells cultured in vitro: the effect of vitamin E succinate supplementation. AB - Malignant murine melanoma (BL6-F10) cells convert arachidonic acid primarily to PGD2, PGF2alpha, PGE2, PGI2 in descending order of magnitude. Supplementation with 1-10 microg/ml vitamin E succinate resulted in a significant (P < or = 0.05) decrease in PGD2 levels at vitamin concentrations of 3, 5, 7 and 10 microg/ml respectively, while PGF2alpha levels were significantly decreased at 1, 3, 5 (P < or = 0.05), 7 and 10 microg/ml (P < or = 0.01) vitamin E succinate. BL6-F10 cells supplemented with 7 and 10 microg/ml vitamin E succinate showed a marked increase in PGE2 levels with a significant increase occurring at 10 microg/ml (P < or = 0.025). PGI2 levels followed a similar trend to PGE2 with a significant increase (P < or = 0.05) occurring at 10 microg/ml. PMID- 9223657 TI - Effects of 17 beta estradiol on the metabolism of labelled arachidonic acid in uteri isolated from intact and ovariectomized rats. Influence of a restricted diet. AB - The effects of restricted diet (50% of the normal intake during 25 days) on the metabolism of 14U arachidonic acid, were explored in uterine horn strips isolated from intact and ovariectomized rats, treated by 17 beta-estradiol or controls. The metabolism of arachidonic acid into different eicosanoids, PGE2, PGF2alpha, 6 keto PGF1alpha and TXB2, showed that the restricted diet diminished PGE2 and PGF2alpha, in intact rats, significantly. In contrast, this kind of feeding did not produce any change in castrated rats. Tissue preparations from previously estrogenized intact and castrated normal-fed rats showed that the production of different metabolites decreased. A similar result was obtained in intact rats subjected to a restricted diet. Nevertheless, in castrated underfed rats, estrogens did not produce any effect on the various eicosanoids analysed. These results showed that in isolated uteri, the effects of 17 beta-estradiol, on metabolite production from labelled arachidonic acid, are different from controls in ovariectomized diet-restricted rats. PMID- 9223658 TI - Inhibition of growth in oral squamous carcinoma cells by cyclopentenone prostaglandins: comparison with chemotherapeutic agents. AB - Four cyclopentenone prostaglandins (CPPGs) and PGE2 caused significant dose dependent inhibition in growth of human oral squamous carcinoma cells (SCC-15). The rank order of their potency was PGJ2>PGA1>16, 16-dimethyl PGA1>PGA2>PGE2. In a follow-up experiment it was found that the mean per cent inhibition in cell growth by PGJ2 and delta12-PGJ2 at 10(-5) M was 61.22 and 63.81, while that of 5 fluorouracil and methotrexate was 36.67 and 38.86, respectively. delta12-PGJ2 and PGJ2 induced significant dose-dependent inhibition in nuclear DNA synthesis (i.e. cell proliferation). Combining vitamin E succinate with lower concentrations of CPPGs enhanced significantly their inhibitory effect on nuclear DNA synthesis of cancer cells. PMID- 9223659 TI - Mechanism of thrombin-induced arachidonic acid release in osteoblast-like cells. AB - In a previous study, we have reported that thrombin stimulates phosphatidylcholine hydrolysis by phospholipase (PL) D, but has little effect on phosphoinositide hydrolysis by PLC in osteoblast-like MC3T3-E1 cells. In the present study, we investigated the mechanism of the thrombin-induced arachidonic acid (AA) release in MC3T3-E1 cells. Thrombin stimulated AA release dose dependently in the range between 0.1 and 1 U/ml. Quinacrine, a PLA2 inhibitor, suppressed the thrombin-induced AA release. In addition, quinacrine also suppressed the thrombin-induced prostaglandin E2 synthesis in these cells. On the other hand, propranolol, which is known to inhibit phosphatidic acid phosphohydrolase, did not affect the thrombin-induced AA release. 1(6-((17beta-3 Methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H- pyrrole-2,5-dione (U-73122), a PLC inhibitor, had no effect on the AA release by thrombin. In addition, 1,6 bis-(cyclohexyloximinocarbonylamino)-hexane (RHC-80267), a selective inhibitor of diacylglycerol lipase, had little effect on the thrombin-induced AA release. Neither propranolol, U-73122 nor RHC-80267 affect the thrombin-induced prostaglandin E2 synthesis. These results strongly suggest that thrombin induces AA release not by phosphatidylcholine hydrolysis by PLD nor phosphoinositide hydrolysis by PLC but mainly by PLA2 in osteoblast-like cells. PMID- 9223660 TI - Protective effects of a thromboxane synthetase inhibitor and continuous arteriovenous hemofiltration in rat endotoxic shock. AB - We determined the efficacy of continuous arteriovenous hemofiltration (CAVH) and a thromboxane synthetase inhibitor (TSI) on survival and their effect on TXA2, PGI2, TNF alpha, and IL-1beta production in rat endotoxemia. Thirty-six endotoxemic rats were randomized to one of 4 groups: (A) no TSI, sham CAVH; (B) no TSI, CAVH; (C) TSI, sham CAVH; and (D) TSI, CAVH. Either CAVH (Group B) or pretreatment with TSI (Group C) resulted in increased survival time. CAVH did not prevent the rise in TX (Group B). TNF alpha levels at 2 h after LPS infusion were higher in Group D compared to Group B (26.1 +/- 3.7 vs 13.2 +/- 4.3 ng/mL, P < 0.05) respectively. IL-1beta was detected earlier in Groups C,D when compared to Groups A,B (P < 0.02). TNF alpha and IL-1beta were not ultrafiltered. CAVH and the inhibition of TX synthesis independently improved survival in endotoxemia, however, their beneficial effects were not additive. While TSI may improve survival by blocking TXA2 production, the salutary effects of CAVH appear to be from removal of an undetermined TXA2 dependent mediator. TNF alpha and IL-1beta concentrations do not appear to influence survival times in this model. PMID- 9223661 TI - Fatty acid composition, eicosanoid production and permeability in skin tissues of rainbow trout (Oncorhynchus mykiss) fed a control or an essential fatty acid deficient diet. AB - Rainbow trout (Oncorhynchus mykiss) were fed either a control diet containing fish oil or an essential fatty acid (EFA) deficient diet containing only hydrogenated coconut oil and palmitic acid as lipid source (93.4% saturated fatty acids) for 14 weeks and the fatty acid compositions of individual phospholipid classes from skin and opercular membrane (OM) determined. The permeability of skin and OM to water and the production of eicosanoids in skin and gills challenged with the Ca2+ ionophore A23187 were also measured. Phospholipid (PL) fatty acid compositions were substantially modified in EFA-deficient fish, with increased saturated fatty acids and decreased polyunsaturated fatty acids (PUFA), especially arachidonic acid (AA) and eicosapentaenoic acid (EPA), while docosahexaenoic acid (DHA) was largely retained. The onset of EFA deficiency was shown by the appearance of n-9 PUFA, particularly 20:3n-9. The main effects of EFA deficiency on phosphatidylcholine (PC) and phosphatidylethanolamine (PE) were to increase saturated fatty acids and monoenes, especially 16:1 and 18:1, and to decrease EPA and DHA. The content of DHA in phosphatidylserine (PS) was high in control animals (40% in skin and 35% in opercular membrane) and was mostly retained in EFA deficient animals. Arachidonic acid (AA) was the most abundant PUFA esterified to phosphatidylinositol (PI) and was significantly reduced in EFA deficient animals (from 31% to 13% in skin), where a large amount of 20:3n-9 (9% in skin) was also present. Influxes and effluxes of water through skin and opercular membrane were measured in vitro. No differences were detected between rainbow trout fed the control or the EFA deficient diet. 12 Hydroxyeicosatetraenoic acid (12-HETE), 12-hydroxyeicosapentaenoic acid (12-HEPE) and 14-hydroxydocosahexaenoic acid (14-HDHE) could not be detected in skin from control or EFA deficient fish. There was no difference between control and EFA deficient trout in the levels of leukotriene C4 (LTC4) and leukotriene C5 (LTC5) in skin cells challenged with the calcium ionophore A23187, and of prostaglandin F2alpha (PGF2alpha), 12-HETE and 12-HEPE in gill cells challenged similarly. Prostaglandin F3alpha (PGF3alpha) production by ionophore stimulated gill cells was significantly reduced in fish fed the EFA-deficient diet. 14-HDHE produced by gill cells was 3.3 fold higher in EFA deficient fish compared to controls. PMID- 9223662 TI - A human papillomavirus type 18 E6/E7 transgene sensitizes mouse lens cells to human wild-type p53-mediated apoptosis. AB - We have studied the concerted action of factors that influence the balance between cell proliferation and cell death in the developing lens of transgenic mice. We show that a human papillomavirus type 18 (HPV18) E6/E7 transgene that predominantly expresses the viral E7 gene product triggers apoptosis in a dose dependent manner, and causes retardation of lens growth or microphakia. E7 is known to inactivate pRB, the product of the retinoblastoma gene, and to enhance the action of p53. Our earlier work had demonstrated that over-expression of p53 itself can cause apoptosis of lens cells, and that a mutant p53 allele can interfere with this process. In the present study, we examined lenses that simultaneously express different constellations of the HPV18 E6/E7, wild-type and mutant human p53, and wild-type human pRB transgenes. We observed that lens cells expressing the HPV18 transgene are more sensitive to wild-type human p53 action than normal lens cells. As a result, there is severe microphakia in lenses that express both the HPV18 and the wild-type p53 transgenes. By contrast, apoptosis was reduced in lenses that co-expressed the HPV18 and either the pRB or the mutant p53 transgene. We conclude that levels of wild-type p53 are critical, and that any excess of p53 or suppression of pRB can cause cell death. Our results encourage attempts to counteract the deleterious action of human papillomaviruses in cervical cancer by a combination of measures that decrease cell proliferation and enhance apoptosis. PMID- 9223663 TI - Suppression of autocrine cell proliferation and tumorigenesis of human melanoma cells and fibroblast growth factor transformed fibroblasts by a kinase-deficient FGF receptor 1: evidence for the involvement of Src-family kinases. AB - Basic Fibroblast Growth Factor (bFGF/FGF2) is thought to play a decisive role in malignant progression. Aberrant expression of bFGF causes constitutive autocrine activation of its cognate receptor and autonomous growth of human melanoma cells or bFGF transformed fibroblasts in culture. It remains to be determined, however, whether the endogenous bFGF confers growth advantage to tumors and what are the downstream targets of the activated FGF receptor critical for its transforming capacity. We therefore transfected metastatic melanoma cells and bFGF transformed mouse fibroblasts with a dominant-negative mutant of the murine FGF receptor 1 (fgfr1/flg), comprising the extracellular and transmembrane domains but lacking the intracellular kinase domain (dnflg). Reverse transcriptase-PCR, 125I-bFGF binding and affinity labeling analyses show that the truncated receptor is targeted to the membrane and is expressed at much higher levels than the endogenous receptor in all of the selected clones. Expression of the dnflg dramatically reduces the basal as well as bFGF induced growth of these cells in vitro and also suppresses their tumorigenic potential in nude mice. The expression of the dnflg does not significantly alter the general level of tyrosyl phosphorylated proteins in the trunsduced melanoma cells. Rather, a major downstream affected target is a Src-family kinase, whose activity, determined by an in vitro immune kinase assay, is stimulated in normal melanocytes by exogenous bFGF, and is markedly reduced in the dnflg-expressing melanoma cells. The present study demonstrates that direct interference with the activity of FGF receptors has a deleterious effect on cell proliferation and survival in vitro and in vivo leading to the suppression of melanoma tumor progression possibly through the inactivation of a Src-family kinase. PMID- 9223664 TI - Chromosomal translocations deregulating c-myc are associated with normal immune responses. AB - Plasmacytomas induced in BALB/c mice by pristane consistently evidence chromosomal translocations involving the c-myc gene and one of the Ig loci. This observation has lead to the suggestion that c-myc deregulation is a critical event in the generation of such tumors. However, it is not clear whether c-myc translocation is related to pristane treatment or occurs in normal lymphocyte populations nor whether such translocations occur normally, and at similar frequencies, in strains genetically resistant to plasmacytoma development, such as DBA/2. In order to address these questions, a Long Distance PCR assay with single copy sensitivity was employed to assess the frequency of c-myc/IgA translocations in normal and immunized mice of both plasmacytoma resistant and susceptible lineages in the absence of pristane treatment. Our data demonstrate that spontaneous translocations occur in normal DBA/2 and BALB/c mice with no significant differences in frequency. A 3-5-fold increase in translocation frequency was observed in mice immunized with cholera toxin, a strong stimulator of IgA responses. We conclude that c-myc deregulation by chromosomal translocation is associated with normal physiological processes of B-cell differentiation and, as such, can not be the determining factor leading to malignancy. PMID- 9223665 TI - Serum starved v-mos-transformed cells are unable to appropriately downregulate cyclins and CDKs. AB - Serum deprived v-mos-transformed NIH3T3 cells are unable to enter a true quiescent state, but instead, arrest in the early G1 phase of the cell cycle. We have analysed several cell cycle regulatory proteins in these G1 arrested cells and show altered regulation in the expression and activity of certain cyclins and cyclin-dependent kinases. In particular, p34cdc2, cyclin A, cyclin D and cyclin E are not appropriately down-regulated in serum starved, G1 arrested, v-mos transformed cells as compared with quiescent NIH3T3 cells. Furthermore, serum starved v-mos-transformed cells have elevated histone H1 kinase activity associated with cyclin A, cyclin E, p33cdk2, and p34cdc2. Using a metallothionein inducible c-mos(mu) expression system, we show that c-mos(mu) induction in quiescent NIH3T3 cells causes elevated expression of p34cdc2. However, this induction of c-mos(mu) and subsequent expression of p34cdc2 was not sufficient to promote significant entry of cells into S phase. Analysis of extracts from serum starved v-H-ras, v-src, and tpr-met transformed NIH3T3 cells demonstrates that these oncogene-transformed cells also contain elevated levels of p34cdc2. We propose that the altered regulation of these critical cell cycle regulatory molecules, and specifically the inability to fully downregulate their activity, contributes significantly to neoplastic transformation and subsequent unregulated growth of tumor cells. PMID- 9223666 TI - Deregulation of specific E2F complexes by the v-mos oncogene. AB - The product of the c-mos proto-oncogene is a protein kinase that is normally expressed in germ cells and functions during oocyte maturation. It has been shown, however, that inappropriate expression of either the viral or cellular mos gene can induce neoplastic progression in somatic cells. Furthermore, v-mos transformed NIH3T3 cells will undergo arrest of proliferation in early G1 upon serum withdrawal but are unable to appropriately down-regulate cell cycle regulatory proteins, such as cyclin and cdc2 proteins, that normally are down regulated in quiescent, untransformed NIH3T3 cells. Since the levels of these proteins are partially transcriptionally controlled, we investigated whether there were alterations in the expression of E2F and AP-1 transcription factor complexes. Indeed, the putative G0/G1-specific p130-E2F complex that is normally observed during low serum-induced cell cycle arrest in NIH3T3 cells is not present in serum starved v-mos-transformed cells. Instead, G1-phase arrested v mos-transformed cells stably express two E2F protein complexes that are normally observed only during S-phase in untransformed cells. The elevation of these complexes in arrested v-mos-transformed cells may be the cause of the transcriptional activation of the E2F-regulated genes cdc2, DHFR, cyclin A, and E2F1 seen in serum starved v-mos-transformed cells. In addition, there are high levels of AP-1 DNA binding activity in serum starved v-mos-transformed cells compared to very low amounts in nontransformed cells. This altered regulation of transcription factor complexes and cell cycle control proteins upon serum withdrawal may provide a mechanism for the uncontrolled cell growth associated with neoplastic transformation induced by certain proto-oncogenes. PMID- 9223667 TI - Transcriptional regulation of the hepatocyte growth factor (HGF) gene by the Sp family of transcription factors. AB - In this study, we have localized an enhancer element in the 5'-flanking region of the HGF gene promoter and have identified its functional transcriptional factors. Through transient transfection of NIH3T3 fibroblast cells and gel mobility shift assays, the functional binding site was localized to a short region (-318 to -303 bp from the transcription start site) which has a CTCCC sequence. This motif is also conserved in the human HGF promoter and acts as a binding site for the Sp family of transcription factors. In the presence of NIH3T3 nuclear protein extracts, this enhancer region formed specific DNA-protein complexes which were totally abrogated by excess amounts of consensus Sp1 oligonucleotide. In gel mobility supershift assays, anti-Sp1 and anti-Sp3 antibodies specifically recognized these complexes as was evident by their slower migration. Site specific mutation of this binding motif resulted in total loss of Sp1 and Sp3 binding and a concomitant loss of enhancer function within the context of the HGF promoter. Furthermore, in transient cotransfection assays, overexpression of Sp1 and/or Sp3 stimulated HGF promoter activity independently and additively through binding to the Sp1 binding site in the HGF gene promoter region. DNaseI hypersensitive analysis of freshly isolated nuclei from NIH3T3 cells revealed that five hypersensitive sites (HSS) are present within the 2 kb region immediately upstream of the HGF promoter. One of these sites was mapped to position -0.3 kb from the transcription start site. These data show that both Sp1 and Sp3 transcription factors upregulate HGF promoter activity by binding to the Sp1 binding site at position -318 to -303 bp in the HGF gene promoter. PMID- 9223669 TI - Frequent loss of heterozygosity on chromosome 10q in muscle-invasive transitional cell carcinomas of the bladder. AB - Loss of heterozygosity (LOH) on chromosome 10 has been observed in several human cancers including glioblastomas, meningiomas, melanomas and endometrial and prostate carcinomas. We have investigated the incidence of LOH on chromosome 10 in 36 human transitional cell carcinomas (TCCs) of the bladder, three upper urinary tract TCCs and one lymph node metastasis, using a panel of 27 highly polymorphic markers spanning 10p (short arm) and 10q (long arm). Fourteen bladder tumours (39%), the three upper urinary tract tumours and the lymph node metastasis showed LOH for at least one locus on chromosome 10. Remarkably, LOH on chromosome 10 was observed mainly in muscle-invasive (P = 0.01) and high grade tumours (P = 0.03). For five tumours and the lymph node metastasis, LOH was found at all informative loci, indicating monosomy or isodisomy of chromosome 10. The deletion mapping of the tumours with partial loss delineated two minimal regions of loss on chromosome 10q. One region, the most telomeric, was bounded by markers D10S214 and D10S169 and the other, the most proximal, was bounded by markers D10S222 and D10S531. Our results demonstrate that chromosome 10q LOH is common in muscle-invasive bladder cancers and that two potential tumour suppressor loci, at 10q24.1-q24.3 and 10q26.1-q26.2, may contribute to the malignant progression of these tumours. Localization of the smallest common regions of loss in bladder tumours provides a starting point for the identification of the genes involved. PMID- 9223668 TI - Csa-19, a radiation-responsive human gene, identified by an unbiased two-gel cDNA library screening method in human cancer cells. AB - A novel polymerase chain reaction (PCR)-based method was used to identify candidate genes whose expression is altered in cancer cells by ionizing radiation. Transcriptional induction of randomly selected genes in control versus irradiated human HL60 cells was compared. Among several complementary DNA (cDNA) clones recovered by this approach, one cDNA clone (CL68-5) was downregulated in X irradiated HL60 cells but unaffected by 12-O-tetradecanoyl phorbol-13-acetate, forskolin, or cyclosporin-A. DNA sequencing of the CL68-5 cDNA revealed 100% nucleotide sequence homology to the reported human Csa-19 gene. Northern blot analysis of RNA from control and irradiated cells revealed the expression of a single 0.7-kilobase (kb) messenger RNA (mRNA) transcript. This 0.7-kb Csa-19 mRNA transcript was also expressed in a variety of human adult and corresponding fetal normal tissues. Moreover, when the effect of X- or fission neutron-irradiation on Csa-19 mRNA was compared in cultured human cells differing in p53 gene status (p53-/- versus p53+/+), downregulation of Csa-19 by X-rays or fission neutrons was similar in p53-wild type and p53-null cell lines. Our results provide the first known example of a radiation-responsive gene in human cancer cells whose expression is not associated with p53, adenylate cyclase or protein kinase C. PMID- 9223670 TI - The phosphatidylinositol 3' kinase pathway is required for the survival signal of leukocyte tyrosine kinase. AB - Leukocyte tyrosine kinase (LTK) is a receptor tyrosine kinase which belongs to the insulin receptor superfamily and is mainly expressed in pre-B lymphocytes and neuronal tissues. Recently, we demonstrated that LTK utilizes Shc and IRS-1 as two major substrates and while both equally activate the Ras pathway, only IRS-1 suppresses apoptosis of hematopoietic cells, suggesting the existence of another unidentified signaling pathway downstream of IRS-1, which is relevant to the anti apoptotic activity. In the present study, we found that wortmannin, a specific inhibitor of phosphatidylinositol 3' (PI3)-kinase, abolished the survival effects of LTK. Although c-Cbl is found to be phosphorylated by LTK and therefore is a second candidate linking LTK with the PI3-kinase pathway along with IRS-1, we found that the p85 subunit of PI3 kinase directly binds to tyrosine 753 of LTK, which is located within a YXXM motif, a consensus binding amino acid sequence for the SH2 domain of p85, but fails to bind to IRS-1 or c-Cbl. Ba/F3 cells which stably express the EGF receptor-LTK chimeric receptor carrying a mutation at tyrosine 753 fell into apoptotic death even in the presence of EGF, indicating that the PI3 kinase pathway is required for the survival effects of LTK. PMID- 9223671 TI - Establishment of a monkey kidney epithelial cell line with the BARF1 open reading frame from Epstein-Barr virus. AB - We previously reported that the BARF1 (BamH1-A right frame 1) gene product from Epstein-Barr Virus (EBV) may have oncogenic properties since injection into new born rats of transfected cell lines resulted in the development of BARF1 expressing tumors, which were aggressive in the case of murine fibroblasts and transient in that of human B lymphocytes. As EBV has been associated with nasopharyngeal carcinoma (NPC) and evidence of BARF1 transcription in this cancer was emerging from our biopsy analyses, we examined the effects of BARF1 transfection into primate primary epithelial cells. The expression of the BARF1 open reading frame in primary monkey kidney epithelial cells led us to the establishment of continuously dividing lines. The BARF1 transfectants showed the major characteristics of immortalized cells: morphological change, short cell doubling time, ability to divide at low cell density and continuous growth over 50 passages. Injection of BARF1 transfectants into nude mice did not induce any tumor. Established subclones were shown to be epithelial cells expressing known keratins as well as the BARF1 coded mRNA and protein. This is the first report indicating that expression of the BARF1 gene product in primary epithelial cells may contribute to the establishment of cell lines. PMID- 9223672 TI - CBP/p300 as a co-factor for the Microphthalmia transcription factor. AB - The Microphthalmia basic-Helix-Loop-Helix-Leucine Zipper (bHLH-LZ) transcription factor (Mi) plays a crucial role in the genesis of melanocytes; mice deficient for a functional (Microphthalmia) gene product lack all pigment cells. We show here that the Mi activation domain resides N-terminal to the DNA-binding domain and that as little as 18 amino acids are sufficient to mediate transcription activation. The minimal activation region of Mi is highly conserved in the related transcription factor TFE3 and is predicted to adopt an amphipathic alpha helical conformation. This region of Mi is also highly conserved with a region of E1A known to be essential for binding the CBP/p300 transcription cofactor. Consistent with these observations, the Mi activation domain can interact in vitro with CBP specifically through a region of CBP required for complex formation with E1A, P/CAF and c-Fos, and anti p300 antibodies can co immunoprecipitate Mi from both melanocyte and melanoma cell lines. In addition, co-transfection of a vector expressing CBP2 (aas 1621-1891) fused to the VP16 activation domain potentiated the ability of Mi to activate transcription, confirming the significance of the CBP-Mi interaction observed in vitro. These data suggest that transcription activation by Mi is achieved at least in part by recruitment of CBP. The parallels between transcription regulation by Microphthalmia in melanocytes and MyoD in muscle cells are discussed. PMID- 9223674 TI - The familial Wilms' tumour susceptibility gene, FWT1, may not be a tumour suppressor gene. AB - A familial Wilms' tumour susceptibility gene, known as FWT1, has recently been localised to chromosome 17q12-q21 by genetic linkage analysis. Four Wilms' tumours from a family showing strong evidence of linkage to FWT1 were examined for allele loss using polymorphic microsatellite markers on chromosome 17q. In three tumours no loss of heterozygosity was observed. In the remaining case, loss of heterozygosity was detected at all markers analysed. However, the alleles lost in this Wilms' tumour were those segregating with the disease in the family. This is in contrast to the usual pattern observed in familial cancer syndromes, where the allele lost in tumours arising in gene carriers is the wild type inherited from the non mutation carrying parent. Taken together with previous data indicating that LOH on chromosome 17q is rare in sporadic Wilms' tumour, the results suggest that FWT1 is not a tumour suppressor gene. Moreover, loss of alleles linked to the disease and the implied absence of the mutated susceptibility gene in one tumour, suggests that a mutation in FWT1 may be necessary for the initiation of some familial Wilms' tumours but subsequently the maintenance of the neoplastic phenotype becomes independent of the FWT1 mutation. PMID- 9223673 TI - Immortalization of neuro-endocrine cells from adrenal tumors arising in SV40 T transgenic mice. AB - Pheochromocytomas are adrenal medullary tumors which arise from the transformation of neural crest-derived cells. In the course of studies of mice transgenic for an SV40 T-gene ectopically expressed in the adrenal medulla, we observed the occurrence of large, mainly bilateral tumors in a high proportion of transgenic animals. From these tumors we established immortalized cell lines which grow in vitro at 32 degrees C (the permissive temperature for the tsA58 T protein encoded by the transgene), but not at 38 C. These cells demonstrate characteristics of both neuronal (160 kd neurofilament) and endocrine (chromogranins) cells. The expression of Mash-1 and ret supports their initial characterization as early bipotential neuro-endocrine progenitors. PMID- 9223675 TI - Novel germline RET proto-oncogene mutations associated with medullary thyroid carcinoma (MTC): mutation analysis in Japanese patients with MTC. AB - Germ-like and somatic mutations in the RET proto-oncogene are associated with inherited and sporadic medullary thyroid carcinoma (MTC). The majority of patients with multiple endocrine neoplasia type 2A (MEN2A) and familial medullary thyroid carcinoma (FMTC) carry germ-line point mutations that result in the substitution of one of five cysteine residues. We investigated exons 10, 11, 13, 14 and 16 of the RET proto-oncogene in 33 unrelated Japanese patients with MTC. Eleven of the 33 cases (33%) were found to have germ-line mutations. Three previously unreported mutations in exon 10 and 11 were identified: one in codon 620, (TGC-->GGC), resulting in a cysteine to glycine substitution, and two in codon 630, (TGC-->TCC) and (TGC-->TAC), resulting in cysteine to serine and cysteine to tyrosine changes, respectively. The new mutations were present in the germ-line DNA of four unrelated patients for whom a family history of MTC had not been documented. Because the new RET alleles described here involve cysteine residues in a region of protein previously associated with FMTC and MEN2A, it is very likely that they represent mutations that predispose to the development of MTC. PMID- 9223676 TI - Co-ordinated regulation of the plasma membrane calcium pump and the sarco(endo)plasmic reticular calcium pump gene expression by Ca2+. AB - The plasma membrane calcium pump (PMCA) and sarco(endo)plasmic reticular calcium pump (SERCA) are important components of the Ca2+ homeostasis system responsible for intracellular Ca2+ signaling, yet the factors which govern their expression remain unclear. Recently, we have found that overexpression of PMCA by a gene transfection approach caused a down-regulation of the SERCA pump [Liu B.F., Xu X., Fridman R., Muallem S., Kuo T.H. Consequences of functional expression of the plasma membrane calcium pump isoform 1a. J Biol Chem 1996; 271: 5536-5544]. The results suggest an interdependence between PMCA and SERCA gene expression which has prompted us to investigate further on the mechanisms that regulate the expression of these Ca2+ pump genes in various cultured cell lines. In the present study, we have analyzed the isoforms of the PMCA and SERCA in different cells and presented evidence in favor of a co-induction of the PMCA and SERCA gene expression by second messengers, such as protein kinase C, cAMP, and Ca2+. Ectopic expression of PMCA or SERCA led to downregulation of the endogenous forms of both pumps. Changes in the level of mRNAs were paralleled by corresponding altered pump protein contents. The Ca(2+)-mediated increase of gene expression is accompanied by increased transcription rates as indicated by nuclear run-on assay. The co-ordinated induction of the PMCA and SERCA gene expression by thapsigargin was not blocked by the cytosolic application of the Ca2+ chelator BAPTA. We conclude that genes encoding components of the major Ca2+ transport pathways, including pumps and IP3 receptor channels, are regulatorally linked and this link is provided by the Ca2+ load of the ER store. This study points to the importance of gene expression as an integral component in the regulation of cellular Ca2+ homeostasis. PMID- 9223677 TI - Bergmann glial cells in situ express endothelinB receptors linked to cytoplasmic calcium signals. AB - The endothelin (ET) isoforms ET-1, ET-2 and ET-3 applied at 100 nM triggered a transient increase in [Ca2+]i in Bergmann glial cells in cerebellar slices acutely isolated from 20-25 day-old mice. The intracellular calcium concentration ([Ca2+]i) was monitored using Fura-2-based [Ca2+]i microfluorimetry. The ET triggered [Ca2+]i transients were mimicked by ETB receptor agonist BQ-3020 and were inhibited by ETB receptor antagonist BQ-788. ET elevated [Ca2+]i in Ca(2+) free extracellular solution and the ET-triggered [Ca2+]i elevation was blocked by 500 nM thapsigargin indicating that the [Ca2+]i was released from InsP3-sensitive intracellular pools. The ET-triggered [Ca2+]i increase in Ca(2+)-free solution was shorter in duration. Restoration of normal extracellular [Ca2+] briefly after the ET application induced a second [Ca2+]i increase indicating the presence of a secondary Ca2+ influx which prolongs the Ca2+ signal. Pre-application of 100 microM ATP or 10 microM noradrenaline blocked the ET response suggesting the involvement of a common Ca2+ depot. The expression of ETB receptor mRNAs in Bergmann glial cells was revealed by single-cell RT-PCR. The mRNA was also found in Purkinje neurones, but no Ca2+ signalling was triggered by ET. We conclude that Bergmann glial cells are endowed with functional ETB receptors which induce the generation of intracellular [Ca2+]i signals by activation of Ca2+ release from InsP3-sensitive intracellular stores followed by a secondary Ca2+ influx. PMID- 9223678 TI - Oscillatory Cl- current induced by angiotensin II in rat pulmonary arterial myocytes: Ca2+ dependence and physiological implication. AB - We have previously reported that angiotensin II (ANG II) induces oscillations in the cytoplasmic calcium concentration ([Ca2+]i) of pulmonary vascular myocytes. The present work was undertaken to investigate the effect of ANG II in comparison with ATP and caffeine on membrane currents and to explore the relation between these membrane currents and [Ca2+]i. In cells clamped at -60 mV, ANG II (10 microM) or ATP (100 microM) induced an oscillatory inward current. Caffeine (5 mM) induced only one transient inward current. In control conditions, the reversal potential (Erev) of these currents was close to the equilibrium potential for Cl- ions (Ecl = -2.1 mV) and was shifted towards more positive values in low-Cl- solutions. Niflumic acid (10-50 microM) and DIDS (0.25-1 mM) inhibited this inward current. Combined recordings of membrane current and [Ca2+]i by indo-1 microspectrofluorimetry revealed that ANG II- and ATP-induced currents occurred simultaneously with oscillations in [Ca2+]i whereas the caffeine-induced current was accompanied by only one transient increase in [Ca2+]i. Niflumic acid (25 microM) had no effect on agonist-induced [Ca2+]i responses, whereas thapsigargin (1 microM) abolished both membrane current and the [Ca2+]i response. Heparin (5 mg/ml in the pipette solution) inhibited both [Ca2+]i responses and membrane currents induced by ANG II and ATP, but not by caffeine. In pulmonary arterial strips, ANG II-induced contraction was inhibited by niflumic acid (25 microM) or nifedipine (1 microM) to the same extent and the two substances did not have an additive effect. This study demonstrates that, in pulmonary vascular smooth muscle, ANG II, as well as ATP, activate an oscillatory calcium dependent chloride current which is triggered by cyclic increases in [Ca2+]i and that both oscillatory phenomena are primarily IP3-mediated. It is suggested that ANG II-induced oscillatory chloride current could depolarise the cell membrane leading to activation of voltage-operated Ca2+ channels. The resulting Ca2+ influx contributes to the component of ANG II-induced contraction that is equally sensitive to chloride or calcium channel blockade. PMID- 9223679 TI - Functional equivalence of native light-sensitive channels in the Drosophila trp301 mutant and TRPL cation channels expressed in a stably transfected Drosophila cell line. AB - Drosophila photoreceptors express two putative cation channels encoded by the transient receptor potential (trp) and trp-like (trpl) genes, which represent prototypical members of a novel family of phosphoinositide-regulated calcium influx channels. Mutations of both trp and trpl selectively abolish components of the light-sensitive current and, when heterologously expressed, both generate cation permeable conductances; however, a detailed comparison of recombinant and native channel properties is lacking. To more rigorously test the hypothesis that TRPL channels mediate one component of the light-sensitive current we have generated cell lines (Drosophila S2 cells) stably transfected with trpl cDNA and compared the recombinant channel properties with those of the light-sensitive conductance in situ in a Drosophila trp mutant under identical conditions. We found close correspondence in respect of a number of quantifiable biophysical parameters including: current voltage relationships, ionic selectivity, voltage independent block by external Mg2+ ions and effective single channel conductance and gating kinetics derived by noise analysis. Our estimate of 60-70 pS for channel conductance was confirmed directly in patch clamp recordings of single TRPL channels in S2 cells. These findings indicate that channels encoded by the trpl gene can completely account for the component of the light-sensitive conductance remaining in the trp mutant. PMID- 9223680 TI - A high-resolution, confocal laser-scanning microscope and flash photolysis system for physiological studies. AB - We describe the construction of a high-resolution confocal laser-scanning microscope, and illustrate its use for studying elementary Ca2+ signalling events in cells. An avalanche photodiode module and simple optical path provide a high efficiency system for detection of fluorescence signals, allowing use of a small confocal aperture giving near diffraction-limited spatial resolution (< 300 nm lateral and < 400 nm axial). When operated in line-scan mode, the maximum temporal resolution is 1 ms, and the associated computer software allows complete flexibility to record line-scans continuously for long (minutes) periods or to obtain any desired pixel resolution in x-y scans. An independent UV irradiation system permits simultaneous photolysis of caged compounds over either a uniform, wide field (arc lamp source) or at a tightly focussed spot (frequency-tripled Nd:YAG laser). The microscope thus provides a versatile tool for optical studies of dynamic cellular processes, as well as excellent resolution for morphological studies. The confocal scanner can be added to virtually any inverted microscope for a component cost that is only a small fraction of that of comparable commercial instruments, yet offers better performance and greater versatility. PMID- 9223681 TI - Intracellular calcium chelator BAPTA protects cells against toxic calcium overload but also alters physiological calcium responses. AB - The effect of the membrane-permeant calcium chelator 1,2-bis-(2 aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetra(acetoxymethyl) ester (BAPTA/AM) on ionomycin-induced cellular calcium overload was studied in single differentiated NH15-CA2 neuroblastoma x glioma hybrid cells. To monitor [Ca2+]i we used the fluorescent indicator Fura-2. Preincubation of the cells with 3 microM BAPTA/AM reduced the number of cells showing deregulation of [Ca2+]i during ionomycin-induced calcium influx. The calcium transients elicited by application of KCl were also severely affected by the chelator. These transients, although varying from cell to cell in shape, amplitude and duration, are well reproducible in individual cells. After incubation of cells for 1 h with 0.3-30 microM BAPTA/AM the time course of these cellular transients was markedly slowed. At 1 microM BAPTA/AM, the time constant of decline of [Ca2+]i was increased by a factor of 4.1 +/- 2.4 (n = 14) and the amplitude was reduced to about 50%. With 30 microM BAPTA/AM, the K(+)-induced calcium transients were almost completely inhibited. We conclude that intracellularly loaded calcium chelators may be used for the prevention of [Ca2+]i-induced cell damage, however, at the expense of a disturbed calcium signalling. PMID- 9223682 TI - Okadaic acid induces the release of Ca2+ from intracellular stores in ECV304 endothelial cells. AB - The effects of serine/threonine phosphatase inhibition on endothelial cell cytosolic free Ca2+ ([Ca2+]c) were investigated using okadaic acid and Fura-2 loaded ECV304 endothelial cells. When added to confluent adherent cells, 500 nM okadaic acid induced a transient and oscillatory elevation of [Ca2+]c both in the presence and absence of extracellular Ca2+. In the absence of extracellular Ca2+, depletion of the intracellular Ca2+ stores with either ATP (1 microM) or thapsigargin (100 nM) prevented any further release of Ca2+ on the subsequent addition of okadaic acid. Likewise (in the absence of extracellular Ca2+), a prior release of Ca2+ induced by okadaic acid reduced the magnitude of the response to ATP (1 microM). Taken together these observations indicate that okadaic acid induces Ca2+ release from the agonist-sensitive pool. The okadaic acid-induced Ca2+ release was mimicked by another potent phosphatase inhibitor, calyculin A (10 nM), and also the less potent analogue of okadaic acid, 1-nor okadone (500 nM). The response to okadaic acid was characterised by a series of asynchronous [Ca2+]c oscillations, which at their peak resulted in 40-100% cells, at any one time, having an elevated [Ca2+]c. The response appeared to propagate between adjacent cells and the elevation of [Ca2+]c appeared initially in the cell periphery. In adherent cells, the release of Ca2+ induced by okadaic acid was found to be dependent upon cell density, as the proportion of cells responding to okadaic acid increased as the cell density increased. The response to okadaic acid was not observed in ECV304 cell suspensions. The data suggest that a kinase activity stimulated either directly or indirectly by cell-cell interactions can lead to the release of Ca2+ from the agonist- and thapsigargin sensitive intracellular stores. PMID- 9223683 TI - Emergency medicine: a status report. PMID- 9223684 TI - Workforce projection in emergency medicine: imperfections, but significant implications. PMID- 9223685 TI - Cranial computed tomography for nontrauma patients: critical appraisal of a decision rule. PMID- 9223686 TI - Our basic need for basic science research. PMID- 9223687 TI - Cranial computed tomography in the emergency evaluation of adult patients without a recent history of head trauma: a prospective analysis. AB - OBJECTIVES: To examine the pattern of nontrauma cranial CT use in an urban ED, to identify the rate of significant CT abnormalities in this setting, and to develop criteria for restricting the ordering of CT scans. METHODS: A prospective, observational study of a case series of adults who underwent cranial CT scanning for nontraumatic cases was performed at the EDs of an urban teaching hospital and an affiliated community hospital with a combined annual census of 110,000. Clinically significant CT scans were defined as: 1) acute stroke, 2) CNS malignancy, 3) acute hydrocephalus, 4) intracranial bleeding, or 5) intracranial infection. X2 recursive partitioning was used to derive a decision rule to restrict ordering of CT scans. RESULTS: Only 61 (8%) of 806 CT scans revealed clinically significant abnormalities. The presence of any of the following: age > or = 60 years, focal neurologic deficit, headache with vomiting, or altered mental status, was 100% sensitive (95% CI: 94-100%) and 31% specific (95% CI: 28 33%) in detecting clinically significant CT scans. This set of features had positive and negative predictive values of 11% (95% CI: 8-13%) and 100% (95% CI: 98-100%), respectively. If these criteria had been used to restrict cranial CT use, 229 fewer patients (28%) would have had CT scans obtained and no clinically significant abnormalities would have been missed. CONCLUSION: Clinically significant CT abnormalities were uncommon in this study population, suggesting that current criteria for ordering nontrauma cranial CT scans may be too liberal. In this study, a set of clinical criteria was derived that may be useful at separating patients into high- and low-risk categories for clinically significant cranial CT abnormalities. Before these results are applied clinically, these criteria should be validated in larger, prospective studies. PMID- 9223688 TI - Blood-brain barrier water permeability and brain osmolyte content during edema development. AB - OBJECTIVE: To determine mechanisms that limit changes in brain water content during acute edema development. METHODS: A controlled, laboratory investigation of the physiologic and biochemical correlates of osmotic edema was performed in rats. Hypoosmotic hyponatremia was induced by intraperitoneal injection of distilled water. Serum osmolality and electrolytes and regional blood-brain barrier water permeability. Surface area (P.S) product, osmolyte contents, and capillary size were determined during 120 minutes of hypoosmotic brain edema development. Cerebral water content predicted from these data using a mathematical model of brain water movements was compared with measured changes in brain water content. RESULTS: Fifteen minutes after distilled water injection, mean +/- SEM blood serum osmolality and sodium concentration decreased from 291 +/- 3 mOsm and 131 +/- 13 mmol/L to 267 +/- 3 mOsm and 102 +/- 9 mmol/L, respectively. Specific gravity of cerebral gray matter, cerebral white matter, and basal ganglia decreased throughout the hypoosmotic exposure period and, for gray and white matter, correlated with blood serum osmolality and sodium plus potassium content. Glutamate, but not glutamine, glycine, or taurine, decreased 120 minutes after water injection. The regional water P.S product decreased by 40% to 60% within 60 minutes of the water injection, while capillary diameters in gray and white matter were unchanged. Brain water movements calculated from the mathematical model correctly predicted actual brain water content only if the hydraulic conductivity of the blood-brain barrier was allowed to vary in proportion to the measured P.S product and the measured loss of brain osmolytes was incorporated into the formulation. CONCLUSIONS: During the first hours of hypoosmotic hyponatremia, changes in brain volume are limited by increased resistance to osmotic flux of water into the brain and reduction in the brain content of inorganic and, to a smaller degree, organic osmolytes. PMID- 9223690 TI - The use of antibiotics to prevent serious sequelae in children at risk for occult bacteremia: a meta-analysis. AB - OBJECTIVE: To determine whether antibiotics prevent serious bacterial infections in children at risk for occult bacteremia. METHODS: Meta-analysis of randomized controlled trials involving children aged 3 months to 36 months without a focus of infection and randomized into 2 treatment groups: 1) no antibiotic vs antibiotic or 2) IM ceftriaxone vs oral antibiotic. RESULTS: The use of either an oral antibiotic or IM ceftriaxone did trend toward a reduced risk of serious infection, although neither reached statistical significance (OR = 0.60; 95% CI 0.10, 3.49; and OR = 0.38; 95% CI 0.12, 1.17, respectively). It would be necessary to treat 414 patients to prevent 1 serious bacterial infection. When only children with proven occult bacteremia were analyzed, the use of IM ceftriaxone was statistically significant in preventing serious bacterial infections (OR = 0.25; 95% CI 0.07, 0.89). CONCLUSIONS: Clinical judgment should not be replaced by widespread antibiotic use in the approach to a child with fever. If rapid methods to identify children with occult bacteremia, such as polymerase chain reaction, could be improved and become widely available, then antibiotics could be used judiciously on initial visits. Antibiotic use in all children at risk for occult bacteremia implies the treatment of many children unlikely to benefit from such therapy. PMID- 9223689 TI - Comparative efficacy of hemodialysis and hemoperfusion in severe theophylline intoxication. AB - OBJECTIVE: To determine the comparative efficacy of hemoperfusion and hemodialysis for severe theophylline toxicity in concurrent case series. METHODS: A 10-year, prospective, observational study was performed of consecutive patients referred to a regional poison control center with severe theophylline intoxication in whom either hemodialysis or hemoperfusion was used. The primary outcomes were 1) incidence of major theophylline toxicity (convulsions or life threatening cardiac dysrhythmias) during or after each procedure, 2) calculated theophylline clearance, and 3) procedure-related complications. RESULTS: Over the study period, 56 patients underwent hemodialysis or hemoperfusion as treatment of severe theophylline intoxication. Overall mean age was 40.5 +/- 22.9 years. Mean peak serum theophylline concentration was 103.3 +/- 39.1 micrograms/mL (range 36 to 245 micrograms/mL). Thirty patients (54%) were victims of acute theophylline intoxication, while 18 (32%) had chronic overmedication and 8 (14%) had acute-on therapeutic intoxication. Thirty-nine patients (70%) underwent hemodialysis, while 17 (30%) underwent hemoperfusion. There were no significant intergroup differences in age (39.4 vs 43.0 yr), peak serum theophylline concentration (99.5 vs 112.1 micrograms/mL), time to procedure (8.4 vs 6.3 hr), or duration of procedure (4.1 vs 3.7 hr). Thirty-three percent of the patients undergoing hemodialysis had major toxicity during or after the procedure, compared with 18% of those who received hemoperfusion (p = NS). Post-procedure serum theophylline concentrations were 26.9 vs 30.4 micrograms/mL, corresponding to drug clearance rates of 185.1 and 294.8 mL/kg/hr (p = 0.03). Procedural complications occurred in 3 patients who received hemoperfusion and consisted of bleeding diatheses and hypocalcemia. No complications occurred in patients receiving hemodialysis (p = 0.007). CONCLUSIONS: These data confirm that hemoperfusion provides a higher theophylline clearance rate than hemodialysis. However, hemodialysis appears to have comparable efficacy in reducing the morbidity of severe theophylline intoxication and is associated with a lower rate of procedural complications. PMID- 9223691 TI - Pilot study of cytokines in emergency department patients with systemic inflammatory response syndrome. AB - OBJECTIVE: To determine the potential utility of cytokine and arachidonic acid metabolite levels in ED patients with systemic inflammatory response syndrome (SIRS) as a predictor of progression to severe sepsis. METHODS: A prospective, observational study of test performance was performed using convenience samples of adult control subjects and admitted patients. The latter patients were identified in the ED as having signs of SIRS. Level of cytokines and arachidonic acid metabolites measured from specimens obtained in the ED were compared between groups and associated with the progression of sepsis within 24 hours in the SIRS patients. RESULTS: There were 30 control patients and 29 SIRS patients. There were 8 SIRS subjects who progressed to severe sepsis within 24 hours using the following criteria (hypotension, n = 1; and organ dysfunction, n = 2). Of the 21 SIRS subjects who did not progress to severe sepsis, 11 had resolution of SIRS criteria at 24 hours. There were no significant differences in mean mediator levels between the SIRS patients who progressed to severe sepsis and those who did not. Of the 11 patients with resolution of SIRS criteria at 24 hours, the mean interleukin-6 (IL-6) level was significantly lower than that for the patients who did not recover or who progressed at 24 hours (n = 18); 65.4 +/- 49.1 vs 230 +/- 112 pg/mL, p = 0.001). Six of 15 subjects with IL-6 > 150 pg/mL progressed to severe sepsis (p = NS). Using threshold values based on the range of levels for normals, the sensitivity of an abnormal marker for the development of severe sepsis within 24 hours varied from 50% to 87%, while the specificity varied from 11% to 84%. CONCLUSION: While mean levels were significantly elevated when compared with those of normal control subjects, they had limited ability to predict the subset of patients likely to progress to severe sepsis. However, initial low levels of cytokines may have exclusionary prognostic value. Prospective validation of the latter finding is warranted. PMID- 9223692 TI - New-onset bronchospasm or recrudescence of asthma associated with cocaine abuse. AB - OBJECTIVE: To determine whether the occurrence of new-onset bronchospasm or the recrudescence of asthma is associated with the use of cocaine. METHODS: A consecutive sample of patients presenting to an inner-city adult ED with new onset bronchospasm or recrudescence of bronchospasm after > 5 years were prospectively enrolled in a case-control prevalence study. The bronchospasm patients were queried as to their exposure to illicit drugs, and urine was obtained to screen for cocaine and its metabolite, benzoylecgonine. An age- and sex-matched control group was composed of randomly chosen subjects without respiratory complaints or a history of asthma. The control group was also screened by urine toxicology for cocaine and its metabolite, benzoylecgonine. RESULTS: In the asthma group, 21/59 (36%) had a urine toxicologic screen positive for cocaine metabolite (benzoylecgonine). Of the 21 with a positive screen for cocaine, 8 denied illicit drug abuse. Among the 13 patients reporting drug use, 10 said that they smoked crack and 3 snorted cocaine. In the control group, 8/53 (15%) were positive. Multivariate logistic regression analysis, with adjustment for age and sex, indicated that the use of cocaine was associated with a 3-fold higher prevalence of new-onset bronchospasm or recrudescence of asthma (OR = 3.28, 95% CI: 1.26 to 8.50). CONCLUSIONS: There appears to be an association between cocaine use and new-onset bronchospasm or recrudescence of asthma in this inner-city ED population. Further study is necessary to determine the basis for this association. PMID- 9223693 TI - Differences between chest pain observation service patients and admitted "rule out myocardial infarction" patients. AB - OBJECTIVE: To compare and contrast the patient characteristics of ED patients at low risk for acute cardiac ischemia who were assigned to a chest pain observation service vs those admitted to a monitored inpatient bed for "rule-out acute myocardial infarction" (R/O MI). METHODS: This was a retrospective, cross sectional comparison of adult patients considered at relatively low risk for cardiac ischemia and who were evaluated in 1 of 2 settings: a short-term observation service and an inpatient monitored bed. All patients had an ED final diagnosis of "chest pain," "R/O MI," or "unstable angina" during the 7-month study period. Demographic features and presenting clinical features were examined as a function of site of patient evaluation. RESULTS: Of 531 study patients, 265 (50%) were assigned to the observation service. Younger age (OR = 1.75, 95% CI 1.26, 2.44, for each decrement of 20 years), the complaint of "chest pain" (OR = 2.35, 95% CI 1.34, 4.12), and the absence of prior known coronary artery disease (OR = 1.64, 95% CI 1.13, 2.38) were the principal independent factors associated with assignment to a chest pain observation service bed. CONCLUSIONS: Patients evaluated in a chest pain observation service appear to have different clinical characteristics than other individuals admitted to a monitored inpatient bed for "R/O MI." Investigators should address differences in clinical characteristics when making outcome comparisons between these 2 patient groups. PMID- 9223694 TI - Morningness-eveningness preferences of emergency medicine residents are skewed toward eveningness. AB - OBJECTIVE: To determine the morningness-eveningness (ME) distribution of emergency medicine (EM) residents. METHOD: A voluntary, modified ME questionnaire was administered to all EM residents in the United States at the time of the 1995 American Board of Emergency Medicine's annual In-Training Examination. RESULTS: Seventy-eight percent (2,047/2,614) of the surveys were returned. ME scores ranged from 24 to 76, with a median score of 49 (interquartile range 44, 56). The scores were distributed differently from those of the normal population (p < 0.001), being skewed toward eveningness. There was a correlation (r = 0.13, p < 0.0001) between resident age and ME score, with older residents being more morning-oriented. Males were more morning-oriented than females (p = 0.005), and respondents with children living at home also were significantly more morning oriented (p < 0.001). Stepwise logistic regression showed that the influence of age, gender, and children was cumulative (r = 0.19) but accounted for only 4% of the observed variability. CONCLUSION: EM residents are distributed differently from the normal population in terms of their ME preferences, tending slightly toward eveningness. The importance of this distribution in EM residents in unknown. A longitudinal follow-up of this cohort may help to determine the association of ME preference with overall practice satisfaction, tolerance of shift work, and career longevity. PMID- 9223695 TI - Bilious vomiting in a 9-month-old infant. AB - A case of bilious vomiting in a 9-month-old male is reported. The differential diagnosis of infantile bilious vomiting is reviewed, and appropriate diagnostic studies are discussed. The child was found to have intussusception. The common manifestation of this pediatric surgical emergency may vary considerably from classic descriptions. Bilious emesis in infants must be considered a surgical emergency until proven otherwise; intussusception may manifest as bilious vomiting only. PMID- 9223696 TI - Morganella morganii: a newly reported, rare cause of neonatal sepsis. AB - This case report reviews the clinical course of an 11-day-old boy who developed late-onset neonatal sepsis secondary to a rare neonatal pathogen, Morganella morganii. This gram-negative enteric bacterium, within the Enterobacteriaceae family, has most commonly been a nosocomial pathogen in debilitated, postsurgical patients. Like many other Enterobacteriaceae, M. morganii has an inducible beta lactamase and is resistant to multiple antibiotics. When caring for neonates with culture-proven M. morganii sepsis, the authors recommend administering both a third-generation cephalosporin and an aminoglycoside to ensure that both antibiotics are bactericidal and to reduce the induction of resistance. PMID- 9223697 TI - Structured emergency medicine board review and resident in-service examination scores. AB - OBJECTIVE: To determine whether the institution of a structured board review program is associated with improved in-service examination scores by residents at an emergency medicine (EM) residency program. METHODS: A retrospective, historical control analysis of the association of a board review program and in service examination scores at one EM residency program was performed. A structured board review consisting of monthly reading assignments in "classic" EM textbooks followed by a graded multiple-choice written examination was instituted in 1987. Percentile scores on the American Board of Emergency Medicine (ABEM) in service examination before (1985-1986) and after (1987-1994) initiation of the board review process were compared by resident level (EM-1, 2, or 3). RESULTS: The EM-1 mean percentile score before review was 50.7 and rose to 68.9 after the institution of the board review program (p = 0.039). Mean EM-2 scores (66.8 vs 65.4) and EM-3 scores (74.4 vs 67.4) decreased slightly; these decreases were not statistically significant. Due to the large increase in EM-1 scores, the mean scores for the total program increased slightly (63.4 vs 67.4; p = NS). CONCLUSION: In this study, EM-1 in-service scores improved in association with the institution of a structured board review program. This formalized didactic program may increase the knowledge base and test performance of EM-1 residents. A favorable effect on EM-2 and EM-3 resident scores was not seen. PMID- 9223698 TI - Recurrent near-syncope with flushing. AB - Episodic vasomotor instability with flushing is an uncommon presentation that is suggestive of an endocrine etiology. This report is the case of a 42-year-old woman who presented to the ED 5 times in a 2-week period for recurrent, self limited episodes of light-headedness associated with tachycardia, hypertension, and flushing. The patient's diagnosis eluded detection in both the outpatient and the inpatient settings for several months. The clinical diagnosis was ultimately confirmed by biochemical test samples obtained in the ED during a subsequent symptomatic event. The differential diagnosis of this patient's presentation includes pheochromocytoma, carcinoid syndrome, medullary thyroid carcinoma, systemic mastocytosis, and other endocrine and toxicologic diseases. ED management of the patient with transient yet significant vasomotor changes includes a workup for syncope, initiation of focused biochemical investigations, referral to the appropriate consultant, and consideration for admission. PMID- 9223699 TI - Workforce projections for emergency medicine: how many emergency physicians does the United States need? AB - OBJECTIVE: To mathematically model the supply of and demand for emergency physicians (EPs) under different workforce conditions. METHODS: A computer spreadsheet model was used to project annual EP workforce supply and demand through the year 2035. The mathematical equations used were: supply = number of EPs at the beginning of the year plus annual residency graduates minus annual attrition; demand = 5 full-time equivalent positions/ED x the number of hospital EDs. The demand was empirically varied to account for ED census variation, administrative and teaching responsibilities, and the availability of physician extenders. A variety of possible scenarios were tested. These projections make the assumption that emergency medicine (EM) residency graduates will preferentially fill clinical positions currently filled by EPs without EM board certification. RESULTS: Under most of the scenarios tested, there will be a large deficit of EM board-certified EPs well into the next century. Even in scenarios involving a decreasing "demand" for EPs (e.g., in the setting of hospital closures or the training of physician extenders), a significant deficit will remain for at least several decades. CONCLUSIONS: The number of EM residency positions should not be decreased during any restructuring of the U.S. health care system. EM is likely to remain a specialty in which the supply of board certified EPs will not meet the demand, even at present levels of EM residency output, for the next several decades. PMID- 9223700 TI - Analysis of factors affecting U.S. emergency physician workforce projections. SAEM Workforce Task Force. AB - OBJECTIVES: To use existing data sources to refine prior estimates of the U.S. emergency medicine (EM) workforce and to estimate effects of proposed changes in the U.S. health care system on the EM workforce. METHODS: Relevant data were extracted from the American College of Emergency Physicians (ACEP) 1995 Membership Activity Report, the American Medical Association (AMA) publication "1995/96 Physician Characteristics and Distribution in the U.S.," the American Hospital Association (AHA) 1994 hospital directory, a written survey of each state's medical licensing board and state medical society, and the American Board of Emergency Medicine (ABEM) annual activity report for 1995. These data were used to project workforce supply and demand estimates applicable to workforce models. RESULTS: None of the available information sources had complete data on the number and distribution of emergency physicians (EPs) currently practicing in the United States. Extrapolating the limited reliable statewide EP numbers to make nationwide projections reveals a shortage of EPs needed to fully staff the nation's existing EDs. At least 22 states had an average ratio of < 5 EPs per existing ED. Additional national projections incorporating a decreasing number of U.S. EDs indicate that the current annual number of EM residency graduates will not eliminate the deficit of EPs for at least several decades, given that projected numbers of retiring EPs annually will soon equal the total annual EM residency graduate production. CONCLUSIONS: Although the current data on EPs in practice in the United States are incomplete, the authors project a relative shortage of EPs. More accurate and complete information on the numbers and distribution of EPs in America is needed to improve workforce projections. PMID- 9223701 TI - The threat of funding cuts for graduate medical education: survey of decision makers. AB - OBJECTIVE: To assess the potential actions of medical school deans, graduate medical education (GME) committee chairs, and hospital chief executive officers (CEOs) regarding future funding reductions for residency training. Specifically, institutions with emergency medicine (EM) residencies were surveyed to see whether EM training was disproportionally at risk for reductions. METHODS: An anonymous 2-page survey was used. Ninety-eight EM residency programs were identified using the American Medical Association Graduate Medical Education Directory 1994-95. Seventy deans, 102 GME chairs, and 97 hospital CEOs were identified. The survey posed a hypothetical 25% forced reduction in residency positions and asked the decision makers for their responses. Options included: 1) proportional reductions of training positions from all residencies, 2) proportional reductions in either primary care or specialty residency positions, or 3) reduction or elimination of specific training programs. The survey asked for a first and second choice of residencies to be reduced or eliminated from an alphabetical list of 17. The survey elicited explanations for each program reduction. RESULTS: 200 (74%) of 269 surveys were returned. Eighty-four responders selected specific residencies to be reduced or eliminated. EM was selected 8 times, making EM the seventh most vulnerable residency to be targeted for reductions. The decision makers who selected proportional reductions chose to reduce across all residencies 32 times, among only the specialty residencies 129 times, and among only the primary care residencies 3 times. CONCLUSIONS: In the setting of anticipated residency cuts, favored proportional reductions in specialty residencies would likely affect EM training. However, most GME decision makers with an existing EM residency program do not consider the EM residency a top choice to be reduced or eliminated. PMID- 9223702 TI - Derivation and validation of a methodology for tracking academic stature of medical schools. Task Force on the Development of Emergency Medicine. AB - OBJECTIVE: To derive and validate a methodology for academic ranking of allopathic medical schools in order to track the development of emergency medicine (EM) at academic medical centers. METHODS: A methodology for institutional ranking according to NIH research grant funding was derived by using a well-known multiaxial ordinal ranking of all Liaison Committee on Medical Education (LCME)-accredited allopathic schools in 1990-91 as the criterion standard. This methodology was validated against the same annually updated criterion standard in 1995-96. Both ranking schemes were first stratified by quartile, then aggregated pairwise by contiguous quartiles to develop a 3 x 3 concordance matrix. RESULTS: Ranking by NIH grant funding produced concordance between all 126 schools in the derivation set and 123/124 schools in the validation set. CONCLUSION: NIH research grant funding may be used as a valid methodology for broad categorization of allopathic medical school academic rank. This strategy should therefore serve as a suitable means of following the development of academic EM over time. PMID- 9223704 TI - Fellowship training in emergency medicine? PMID- 9223703 TI - Changing status of academic emergency medicine (1991-1996). Task Force on the Development Emergency Medicine. AB - OBJECTIVE: An SAEM national task force previously concluded that academic departments and residencies in emergency medicine (EM) had preferentially developed outside of the academic mainstream. This study was designed to determine whether EM has made significant inroads into academic medical centers over the past 5 years. METHODS: The baseline data set (7/1/91) contained all 126 Liaison Committee on Medical Education (LCME)-accredited schools, and all 87 Residency Review Committee (RRC)-accredited EM residencies. The comparison data set (7/1/96) contained all 124 LCME-accredited schools, and all 114 RRC accredited EM residencies. The 1991-96 increments in academic departments and university-hospital residencies was examined in the aggregate, then stratified by medical schools grouped into quartiles and contiguous quartiles, according to academic ranking. A-priori and post-hoc comparisons were expressed with 95% and 99% confidence intervals (CIs), respectively. RESULTS: Over the past 5 years, the proportion of academic departments of EM increased by 23%, from 18% to 41% of all LCME-accredited schools (95% CI 12% to 34%). The largest increase (58%; 99% CI 40% to 77%) occurred among those schools academically ranked above the median. The proportion of EM residencies at university hospitals increased by 17%, from 40% to 57% (95% CI 5% to 30%). Again, the largest increase (25%; 99% CI 3% to 47%) occurred at university hospitals affiliated with schools academically ranked above the median. CONCLUSION: EM has made substantial inroads into academic medical centers over the past 5 years. This is reflected in quantitatively and statistically significant increases in academic departments and university hospital residency programs, both occurring largely within institutions whose academic rankings place them among the upper half of all LCME-accredited medical schools. PMID- 9223706 TI - Relief of chest pain out-of-hospital: a lost diagnostic opportunity? PMID- 9223705 TI - Thrombolysis for stroke? PMID- 9223707 TI - Diagnosis of midgut volvulus in an adult with the aid of key radiologic findings. PMID- 9223708 TI - Ileocolic intussusception secondary to a lipoma in an adult. PMID- 9223709 TI - Managing methicillin-resistant Staphylococcus aureus in hospital: the balance of risk. PMID- 9223710 TI - The natural history of delirium in older hospitalized patients: a syndrome of heterogeneity. AB - OBJECTIVES: To determine the presentation, course and duration of delirium in hospitalized older people. DESIGN: Observational cohort study. SETTING: Inpatient surgical and medical wards at a university hospital. PARTICIPANTS: 432 people over the age of 65. MEASUREMENTS: All participants were screened daily for confusion and, in those who were confused, delirium was ascertained using the Diagnostic and Statistical Manual of Mental Disorders (DSM) III-R criteria. Those who were found to be delirious were followed daily while in hospital for evidence of delirium. The Delirium Rating Scale (DRS) was used to describe the clinical characteristics of delirium. RESULTS: About 15% of subjects had delirium. Sixty nine percent of delirious subjects had delirium on a single day. The DRS total was higher on the first day of delirium for those with delirium on multiple days than those with delirium on a single day (P = 0.03). Among those with delirium on multiple days, there were no patterns of change over time in specific DRS items. CONCLUSIONS: Delirium in hospitalized older people is common and has a varied presentation and time course. Clinicians and researchers need to consider this great heterogeneity when caring for patients and when studying delirium. PMID- 9223711 TI - Reliability of parathyroid hormone measurements in the period immediately following hip fracture. AB - AIM: As it is unclear whether parathyroid hormone (PTH) measurements performed immediately after hip fracture are reliable indicators of pre-fracture metabolic status, we set out to define how PTH levels are affected by hip fracture and its surgical repair. METHOD: In two longitudinal projects, we studied 12 patients presenting with hip fracture and eight patients undergoing elective hip replacement. PTH, calcium and 25-hydroxyvitamin D (25OHD) levels were measured on admission, 2 days and 1 week later and after recovery at least 2 months after initial admission. FINDINGS: In the subjects presenting with hip fracture, PTH levels during inpatient care were no different from those subsequently measured during the recovery period. In subjects undergoing elective hip surgery, PTH levels did not change following surgery and again remained unchanged into the recovery period. CONCLUSIONS: Measurements of PTH performed during inpatient care of those with hip fracture appear sufficiently reliable for use in assessment of metabolic status. PMID- 9223712 TI - Longitudinal trends in late-life insomnia: implications for prescribing. AB - OBJECTIVE: To assess trends in insomnia and hypnotic drug use in a representative sample of elderly general practice patients. DESIGN: Longitudinal study with three interview waves--1985, 1989 and 1993. SETTING: Urban and suburban Nottingham. PARTICIPANTS: 1042 patients originally aged 65 and over randomly sampled from general practitioner lists. MAIN OUTCOME MEASURES: Point prevalence estimates, status (case/non-case/died) at 4-year follow-up, episode incidence and survival functions. RESULTS: At baseline (1985) 221 respondents met the survey criteria for insomnia. Of these, 36.1% continued to report severely disrupted sleep in 1989. Within this period 84 new cases of insomnia were identified (an incidence rate of 3.1% per person-year at risk). Controlling for age and sex, insomnia was unrelated to survival among prevalent cases, but significantly related to survival among incident cases (odds ratio = 1.7; 95% confidence interval = 1.1-2.5). Of 166 respondents using prescription hypnotics in 1985, 31.7% continued to report usage in 1989. Similarly, out of 41 new hypnotic drug users identified in 1989, 29.3% continued to report usage in 1993. CONCLUSIONS: Important clinical differences in the natural history of insomnia are evident when incident and prevalent cases are compared. Nevertheless, outcomes at 4-year follow-up suggest that, for the majority of surviving cases identified in a prevalence screen and for a substantial minority of incident cases, late-life insomnia shows a level of chronicity incompatible with hypnotic drug therapy as currently recommended. PMID- 9223713 TI - Sensitivity of the cough reflex in young and elderly subjects. AB - OBJECTIVE: To compare the sensitivity of the cough reflex--which is said to be normal in elderly people--in elderly and young subjects. SUBJECTS: 20 elderly (14 female) subjects, mean (SEM) age 83 (1) years and 20 young (nine female) subjects, mean (SEM) age 27 (1) years, who were all non-smokers. None of the subjects was taking antitussive drugs and none suffered from clinically evident lung, cardiac or neurological disease. Five elderly subjects were unable to perform adequate spirometry and were excluded from analysis. DESIGN AND OUTCOME MEASURES: Each subject inhaled 10 ml of nebulized distilled water and isotonic saline (as placebo) for 30 s, 10 min apart in a randomized double-blind crossover fashion. The cough frequency induced with each treatment was recorded on a click counter. RESULTS: Cough frequency on inhaling distilled water was significantly lower in the elderly group than in the younger group, with a difference of 9.53 (95% confidence intervals: 3.63, 15.4; P < 0.001). None of the subjects coughed when inhaling placebo solution, resulting in significant differences in cough frequencies between distilled water and placebo of 5.87 (2.82, 8.92; P < 0.05) for the elderly group and 15.4 (11.0, 19.8; P < 0.0005) for the younger group. CONCLUSIONS: The sensitivity of the cough reflex appears to be significantly reduced in elderly subjects. This may increase the risk of aspiration and bronchopulmonary infection in old age, even in the absence of respiratory disease. PMID- 9223714 TI - Fear of falling and restriction of mobility in elderly fallers. AB - OBJECTIVES: To identify the characteristics of elderly persons who develop a fear of falling after experiencing a fall and to investigate the association of this fear with changes in health status over time. DESIGN: A prospective study of falls over a 2-year period (1991-92). Falls were ascertained using bimonthly postcards plus telephone interview with a standardized (World Health Organisation) questionnaire for circumstances, fear of falling and consequences of each reported fall. Each participant underwent a physical exam and subjective health assessment each year form 1990 to 1993. SETTING: New-Mexico Aging Process Study, USA. SUBJECTS: 487 elderly subjects (> 60 years) living independently in the community. MAIN OUTCOME MEASURES: Fear of falling after experiencing a fall. RESULTS: 70 (32%) of 219 subjects who experienced a fall during the 2 year study period reported a fear of falling. Women were more likely than men to report fear of falling (74% vs 26%). Fallers who were afraid of falling again had significantly ore balance (31.9% vs 12.8%) and gait disorders (31.9% vs 7.4%) at entry in the study in 1990. Among sex, age, mental status, balance and gait abnormalities, economic resource and physical health, logistic regression analysis show gait abnormalities and poor self-perception of physical health, cognitive status and economic resources to be significantly associated with fear of falling. Subjects who reported a fear of falling experienced a greater increase in balance (P = 0.08), gait (P < 0.01) and cognitive disorders (P = 0.09) over time, resulting in a decrease in mobility level. CONCLUSION: The study indicated that about one-third of elderly people develop a fear of falling after an incident fall and this issue should be specifically addressed in any rehabilitation programme. PMID- 9223715 TI - Environmental hazards in the homes of older people. AB - OBJECTIVES: To investigate (i) the prevalence of environmental safety hazards in the homes of people aged 70 years and over, (ii) their knowledge of causes of injuries to older people and the safety measures they can implement to prevent such injuries and (iii) the relationship between socio-demographic characteristics of this population group and levels of home environmental hazards. METHOD: A cross-sectional survey of 425 people aged 70 years and older living in a defined geographical area of Australia. Participants were recruited through their general practitioners. A structured interview completed with each participant included questions on demographics and home safety issues. A home safety inspection was also undertaken using a predetermined rating format. RESULTS: 80% (n = 342) of homes inspected had at least one hazard and 39% (n = 164) had > 5 hazards. The bathroom was identified as the most hazardous room, with 66% (n = 279) of bathrooms having at least one hazard. Hazards relating to floor surfaces (62% of homes had one 'flooring' hazard) and absence of appropriate grab or handrails (60% of homes had one or more hazards relating to this) were prevalent. Eighty-eight percent (n = 374) of older people were able to identify falls as the most common cause of injury and 87% (n = 368) were able to accurately name at least one safety measure. Although a significant association was found between the older people's self-assessment of their home's safety and the presence of more than 5 hazards, 30% of those rating their homes as very safe (n = 289) had more than 5 hazards. Logistic regression analysis identified one variable--contact with healthcare service providers--as predictive of the hazard level in older people's homes. Older people who were never visited by service providers were twice as likely to have more than 5 hazards as those who were visited weekly or more often (OR 2.12, 95% CI 1.104, 4.088). CONCLUSION: Many older people are living in potentially hazardous environments. As yet, a causal link between the presence of environmental hazards and falls in older people has not been established. More definitive work in this area needs to be carried out. PMID- 9223716 TI - Health status and disability among elderly people in three UK districts. AB - AIM: To obtain population data on health status and disability of elderly people which may help in planning and maintaining services and be used as comparators for research. METHOD: Random samples of people aged 70 and over were interviewed in their own homes in West Glamorgan, Dudley and North Staffordshire. The interviews included standardized assessments of health status (SF-36), disability (Barthel index) and cognitive function (Abbreviated Mental Test). RESULTS: 1608 interviews were completed. Response rates varied between 66 and 84%. Age and sex adjusted scores for five of the eight parameters of the SF-36 and the Barthel score differed significantly between districts. CONCLUSIONS: Local studies are required to provide appropriate normative data for each area. In the absence of such studies, the data in this paper are the best currently available. PMID- 9223717 TI - The role of a specialist team in implementing continuing health care guidelines in hospitalized patients. AB - BACKGROUND: Assessment of continuing health care needs is unstandardized and often undertaken by professionals not trained in the management of complex disability. METHODS: A 6 month prospective study to evaluate the role of a specialist team in implementing continuing care guidelines in hospitalized patients. The team was responsible for assessment and facilitation of access to continuing health care throughout the hospital between hospital and community on a non-age-related basis. It had access to six inpatient beds and a budget to purchase health care after discharge for 7 days. Patients with complex disability were referred to the team if their continuing health care needs could not be assessed, improved or provided within routine practice. RESULTS: Of the 93 patients included in the study, 34 (37%) were from geriatric wards and 59 (63%) from other specialties. Twenty-six (44%) of the patients from other specialties had been inappropriately referred (no continuing health care needs) and 24 (41%) appropriate patients had not been referred because of inadequate assessments. It was possible to facilitate discharge and continuing care provision in 26 patients without transfer to dedicated beds. Thirty-two patients were transferred for further management (median length of stay 17 days). Three (9%) patients died, 20 (63%) were discharged home and six (19%) were discharged to institutional care. Three patients had to be transferred to acute care. A high level of satisfaction with support and post-discharge arrangements was reported by 26 (81%) patients, 25 (78%) carers and 26 (81%) general practitioners for patients transferred to specialist beds. CONCLUSIONS: Specialist intervention, using a team approach, facilitates effective implementation of continuing care guidelines in hospitalized patients. PMID- 9223718 TI - Screening for depression among acutely ill geriatric inpatients with a short Geriatric Depression Scale. AB - BACKGROUND: Depression is not uncommon among acutely ill geriatric inpatients. METHOD: The performances of shorter versions of the Geriatric Depression Scale (GDS) in screening for depression among acutely ill geriatric inpatients were examined. RESULTS: A cut-off of 2/3 gives the best sensitivity (88%) and specificity (75%) for the 10-item version (GDS10). A cut-off of 0/1 gives the best sensitivity (72%) and specificity (90%) for the 4-item version (GDS4). A positive response to item 6 ("Do you often feel helpless?") on the GDS10 gave a sensitivity of 76% and specificity of 75%. Patients found the GDS10 tolerable and acceptable. CONCLUSION: Both shorter versions of the GDS may be utilized in screening for depression among acutely ill geriatric inpatients. PMID- 9223719 TI - The performance of simple instruments in detecting geriatric conditions and selecting community-dwelling older people for geriatric assessment. AB - BACKGROUND: comprehensive geriatric assessment (CGA) appears to be less effective when performed in outpatient clinics than in hospital settings. The effectiveness of outpatient CGA might be improved by selectively targeting frailer community dwelling elderly people. The purpose of this study was to evaluate the clinical performance of rapidly-administered standard screening measures for geriatric syndromes in selecting community-dwelling older people for outpatient CGA. METHODS: urban-dwelling older people were screened for CGA at community sites using a self-administered questionnaire containing standardized measures for each of four geriatric target conditions: depression, urinary incontinence, falls and functional impairment. The study sample included all 150 consecutive subjects who were screened, failed on one or more of the four target criteria and completed community-based, academically administered CGA. Diagnostic accuracy of the screening instruments was assessed using CGA diagnoses as the 'gold standard'. In addition, patients' potential for benefiting from CGA was determined by whether they received major medical recommendations for further evaluation or treatment. RESULTS: after completing CGA, 60.2% of those failing on functional impairment, 53.5% of those failing on depression, 30.7% of those failing on falls and 92.7% of those on urinary incontinence, were confirmed as having these or highly related conditions as clinical problems. Overall, 81.3% of the subjects completing CGA received the least one major recommendations for further medical intervention; most of these recommendations (79.5%) were for a target-related condition and the further remainder (20.5%) addressed another significant medical condition. CONCLUSION: simple screening instruments used in community settings have variable degrees of accuracy, but may be markers for frailty and thus can identify older people likely to benefit from geriatric assessment. PMID- 9223720 TI - A travel account of an 11-city tour of British geriatric units. PMID- 9223721 TI - Longer staying patients on an acute old age psychiatric unit--characteristics and outcome. PMID- 9223722 TI - Macrocytosis in elderly patients. PMID- 9223723 TI - Outcome of elderly patients requiring ventilatory support. PMID- 9223724 TI - Early post-stroke parasitic delusions. PMID- 9223725 TI - Xerostomia: a symptom which acts like a disease. PMID- 9223726 TI - Assessment of well-being. PMID- 9223727 TI - Needle exchange is not enough: lessons from the Vancouver injecting drug use study. AB - OBJECTIVE: To describe prevalence and incidence of HIV-1, hepatitis C virus (HCV) and risk behaviours in a prospective cohort of injecting drugs users (IDU). SETTING: Vancouver, which introduced a needle exchange programme (NEP) in 1988, and currently exchanges over 2 million needles per year. DESIGN: IDU who had injected illicit drugs within the previous month were recruited through street outreach. At baseline and semi-annually, subjects underwent serology for HIV-1 and HCV, and questionnaires on demographics, behaviours and NEP attendance were completed. Logistic regression analysis was used to identify determinants of HIV prevalence. RESULTS: Of 1006 IDU, 65% were men, and either white (65%) or Native (27%). Prevalence rates of HIV-1 and HCV were 23 and 88%, respectively. The majority (92%) had attended Vancouver's NEP, which was the most important syringe source for 78%. Identical proportions of known HIV-positive and HIV-negative IDU reported lending used syringes (40%). Of HIV-negative IDU, 39% borrowed used needles within the previous 6 months. Relative to HIV-negative IDU, HIV-positive IDU were more likely to frequently inject cocaine (72 versus 62%; P < 0.001). Independent predictors of HIV-positive serostatus were low education, unstable housing, commercial sex, borrowing needles, being an established IDU, injecting with others, and frequent NEP attendance. Based on 24 seroconversions among 257 follow-up visits, estimated HIV incidence was 18.6 per 100 person-years (95% confidence interval, 11.1-26.0). CONCLUSIONS: Despite having the largest NEP in North America, Vancouver has been experiencing an ongoing HIV epidemic. Whereas NEP are crucial for sterile syringe provision, they should be considered one component of a comprehensive programme including counselling, support and education. PMID- 9223728 TI - Two parallel routes of the complement-mediated antibody-dependent enhancement of HIV-1 infection. AB - OBJECTIVE: To study the mechanism of the complement-mediated antibody-dependent enhancement (C'-ADE) of HIV infection which may play a significant role in the progression of HIV-disease. METHODS: In vitro complement activating and complement-mediated HIV-infection enhancing abilities of three human anti-gp41 monoclonal antibodies (MAb) were tested. C'-ADE was estimated using HIV-1IIIB and CR2 (CD21)-carrying MT-4 target cells. Normal human serum (NHS), purified C1q, C1q-deficient (C1qD) and C2-deficient (C2D) human sera were applied as complement sources. RESULTS: All MAb mediated increased C1q binding to solid-phase gp41. All MAb had a marked dose-dependent and strictly complement-mediated HIV-infection enhancing effect. Mixtures of the MAb with purified C1q also significantly increased HIV-1 infection. C1qD serum had a markedly lower enhancing effect than NHS, which could be raised to normal level by addition of purified C1q. Pretreatment of the target cells with anti-CR2 antibodies only partially inhibited the enhancing effect of the MAb plus normal human serum. CONCLUSION: These novel findings indicate that besides the well-known facilitation of entry of HIV-1 by the interaction between virus-bound C3 fragments and CR2 present on the target cells, fixation of C1q to intact virions also results in an enhanced productive HIV-1 infection in the MT-4 cell cultures. PMID- 9223729 TI - Characteristics of the CD8+ lymphocytosis during primary simian immunodeficiency virus infections. AB - OBJECTIVE: To investigate the source of the expanded blood CD8+ subsets during an acute primary simian immunodeficiency virus (SIV) infection of macaques and the potential role of these cells in disease progression. DESIGN AND METHODS: The primary CD8+ lymphocytosis, which occurs at 1-2 weeks following infection with SIVsmm/PBj-14, was examined in rhesus and cynomolgus macaques. Extensive subset analysis of the expanded blood CD8+ cell pool in a rhesus macaque was compared phenotypically with those in thymus, lymph nodes, spleen, ileum and lung washouts obtained at necropsy during blood lymphocytosis. The influence of the primary CD8+ cells expansion on disease progression was assessed at days 175-679 post infection in long-term PBj-14 survivors staged according to immunological, virological and histopathological changes in their lymphoid organs. RESULT: The very rapid and transient blood lymphocytosis following infection consisted of two distinct CD45RA(low), CD8+ and CD28-, lymphocyte function-associated antigen (LFA)-1(high), CD45RA(high), CD8+ populations. These populations were present in low levels in thymus, lymph and spleen but were highly represented in mucosal tissues, such as long washout, in which CD28- LFA-1(high) CD45RA(high) CD8+ cells comprised 86% of CD8+ cells, and gut, which was predominantly CD45RA(low) CD28- CD8+ cells. A comparison of progressor and non-progressor PBj-14-infected rhesus and cynomolgus macaques also indicated that the existence or magnitude of a blood CD8+ lymphocytosis during the acute phase of infection did not by itself appear to influence or be predictive of disease progression. CONCLUSION: The marked blood CD8+ lymphocytosis observed during acute SIV infection did not result from expansion of virus-specific precursors in peripheral lymph node and did not appear to influence the rate of disease progression. The findings provide a novel explanation for the primary CD8+ cell lymphocytosis and invoke a mechanism whereby virus-induced cytokine/chemokine production in mucosal sites initiate the transient migration of a pre-existing CD8+ population into the blood from compartments such as lung and gut. Such results suggest that the magnitude of lymphocytosis may depend on the level of viral replication in mucosal tissues and the presence of other infections, for example, cytomegalovirus. PMID- 9223730 TI - Correlation between susceptibility of primary HIV-1 isolates to autologous and heterologous neutralizing antibodies. Hospital Evandro Chagas AIDS Clinical Research Group. AB - OBJECTIVE: To study the susceptibility of primary HIV-1 isolates towards autologous and heterologous neutralizing antibodies (NAb). DESIGN: Blood was collected and primary HIV-1 isolated from individuals residing in Rio de Janeiro, Brazil, in all phases of disease. METHODS: Primary HIV-1 isolates were incubated with autologous or heterologous plasma and neutralization of infection of freshly pre-stimulated normal human peripheral blood mononuclear cells was assayed in parallel to median infectious dose determinations in the absence of antibodies. Levels of HIV-1 p24 antigen were used for evaluation of viral neutralization. RESULTS: Autologous neutralization (75%) was observed for 13 (52%) out of 25 of the primary HIV-1 isolates, and 15 (71%) out of 21 isolates were susceptible to 75% heterologous neutralization by at least one-half of the heterologous plasma tested. Primary HIV-1 isolates susceptible to autologous NAb showed a higher susceptibility towards neutralization by heterologous NAb than isolates that could not be neutralized by the autologous plasma (P = 0.049). The susceptibility of the primary HIV-1 isolates towards neutralization by heterologous NAb was significantly higher for isolates derived from men (P = 0.001), and for isolates obtained from individuals infected through homo-/bisexual risk behaviour in comparison with those infected through heterosexual HIV-1 transmission (P = 0.03). CONCLUSIONS: Susceptibility of primary HIV-1 isolates to autologous and heterologous neutralization was significantly correlated, indicating that escape mutants may become resistant not only to autologous but also to heterologous NAb. PMID- 9223731 TI - Experimental gene therapy: the transfer of Tat-inducible interferon genes protects human cells against HIV-1 challenge in vitro and in vivo in severe combined immunodeficient mice. AB - OBJECTIVES: To evaluate in vitro and in vivo a strategy for gene therapy for AIDS based on the transfer on interferon (IFN)-alpha, -beta and -gamma genes to human cells. DESIGN: Human U937 promonocytic cells were stably transfected with Tat inducible IFN expression vectors conferring an antiviral state against infection with HIV. METHODS: Transfected cells were either infected by HIV-1 in vitro or transplanted into severe combined immunodeficient (SCID) mice for an HIV challenge in vivo. RESULTS: U937 cell lines stably carrying IFN transgenes under the positive control of the HIV-1 Tat protein were highly resistant to HIV-1 replication in vitro. This antiviral resistance was associated with a strong induction of IFN synthesis immediately following the viral infection. HIV-1 proteins were found to be specifically trapped within the genetically modified cells. In contrast, all IFN-U937 cells permitted full HIV-2 replication. Transfected cells injected into SCID mice and challenged against HIV-1 were strongly resistant to infection when cells were transduced with IFN-alpha of IFN beta genes. However, IFN-gamma-transfected cells permitted HIV-1 infection in vivo despite the induction of a high level of IFN-gamma secretion. The quantity of proviral DNA was 10(5)-fold lower in IFN-alpha- or IFN-beta-transfected U937 cells collected from these SCID mice than that in non-transfected cells. CONCLUSIONS: Our results substantiated the validity of a strategy, bases on the transfer of HIV-1-inducible IFN-alpha or IFN-beta genes, to confer antiviral resistance to human cells. PMID- 9223732 TI - Quantification of HIV in semen: correlation with antiviral treatment and immune status. AB - OBJECTIVE: This study examined the concentration of HIV in semen and the effects of biological factors on HIV excretion. METHODS: Semen samples from 101 men at different stages of the disease were evaluated by quantitative HIV culture and HIV RNA detection. Blood plasma samples were available from 56 patients. The effects of CD4 and CD8 count, blood plasma RNA levels, treatment status and clinical staging on the shedding of HIV were evaluated. RESULTS: HIV RNA levels in semen correlated with quantitative HIV culture of seminal cells and a strong association of positive seminal cell culture with high RNA levels was observed. CD4 count and antiviral treatment were inversely correlated with the concentration of HIV in semen. Blood plasma HIV RNA values were correlated with HIV RNA levels in semen, although some patients had highly discrepant results. CONCLUSIONS: The strong correlation between seminal cell culture and concentration of HIV RNA in seminal plasma suggested that HIV detected in seminal plasma was released by productively infected cells in the male genital tract. The study showed that the concentration of HIV in semen, which was likely to be correlated with HIV infectivity, was a function of the immune status of the HIV infected individual. The results suggested that antiviral therapy may have reduced the concentration of HIV in semen. PMID- 9223733 TI - Frequency and risk factors of infectious complications in neutropenic patients infected with HIV. AB - OBJECTIVE: To determine causes, incidence and factors associated with infections in neutropenic [polymorphonuclear neutrophil (PMN), 1000 x 10(6)/l] HIV-infected patients. DESIGN: Prospective study. SETTING: Infectious disease service of a 1000-bed university teaching hospital in Paris, France. PATIENTS: HIV-infected patients with a PMN count of < 1000 x 10(6)/l confirmed on two occasions were included in the study. Baseline characteristics, cause of neutropenia and occurrence of infectious episodes were analysed. RESULTS: The cause of neutropenia was lymphoma in four cases (6.5%), antineoplastic chemotherapy in seven (11.3%), zidovudine in 32 (51%), trimethoprim-sulphamethoxazole (TMP-SMX) in 28 (45%) and ganciclovir in 11 (18%). Fifteen patients (24%) developed infectious complications. Neutropenia induced by chemotherapy or lymphoma was more frequently complicate by infectious episodes (P = 0.02). Neutropenia in the previous 3 months (P = 0.05), presence of a central venous catheter (P = 0.05) and a trough PMN count (P = 0.02) were the three risk factors of infection retained in a logistic model. CONCLUSION: Neutropenia induced by zidovudine, gangiclovir or TMP-SMX, are less complicated by infectious episodes than neutropenia induced by antineoplastic chemotherapy. Overall, infectious episodes in neutropenic HIV-infected patients appear lower than in patients with haemobiologic malignancies. PMID- 9223734 TI - Impact of treatment changes on the interpretation of the Concorde trial. AB - BACKGROUND: The Concorde trial compared two policies of therapy with zidovudine (ZVD) in individuals with asymptomatic HIV infection: immediate or deferred ZDV. Participants in both groups could stop their blinded trial therapy for several reasons and/or could start open-label ZDV. The difference in survival and disease progression between the two groups was estimated allowing for treatment changes. METHODS: The relationship between latest CD4 count, treatment changes and time to AIDS-related complex (ARC), AIDS or death was investigated using time-updated proportional hazards models, but these models gave seriously biased estimates of the effect of ZDV. Therefore, a method based on the comparison of the randomized groups was used. A model relating a participant's events times to the treatment actually received was used to estimate what would have been observed if the deferred group had not received ZDV before ARS or AIDS, and to explore alternative policies for starting Pneumocystis carinii pneumonia (PCP) prophylaxis. RESULTS: The major treatment changes during the trial were the termination of blinded therapy because of adverse events or personal reasons (575 out of 1749 participants), starting open-label ZDV (745 participants), and starting PCP prophylaxis (613 participants). Starting open-label ZDV and PCP prophylaxis were strongly related to latest CD4 count. The uncorrected hazard ratios for immediate compared with deferred groups were 0.89 for time to ARC, AIDS or death [95% confidence interval (CI), 0.75-1.05], 1.01 for time to AIDS or death (95% CI, 0.82-1.24), and 1.26 for time to death (95% CI, 0.93-1.70). After correction for treatment changes, these hazard ratios were 0.79 (95% CI, 0.57 1.11), 1.01 (95% CI, 0.81-1.26), and 1.37 (95% CI, 0.91-2.08), respectively. Correction for PCP prophylaxis made little difference to the results. CONCLUSIONS: Open-label ZDV before ARC or AIDS in the deferred group was likely to have diluted any differences between the immediate and deferred groups. After correction for this dilution, both the estimated benefit of immediate treatment in delaying progression to ARC, AIDS or death and the estimated disadvantage of immediate treatment in accelerating death were somewhat increased, but both remained consistent with chance alone. This study demonstrated the large potential bias inherent in non-randomization-based methods of analysis of clinical trials. PMID- 9223735 TI - Longer survival after HIV infection for injecting drug users than for homosexual men: implications for immunology. AB - BACKGROUND: Comparisons of progression in HIV-1 infection between injecting drug users (IDU) and homosexual men have been inconclusive due to the short follow-up periods, often with less well-defined starting points and endpoints. In addition, comparisons of survival after injection have been to some extent obscured by higher non-AIDS mortality in IDU. METHOD: In a retrospective cohort study, homo /bisexual men and IDU were followed, with dates of seroconversion defined within +/- 1 year by a previously negative HIV antibody test. Endpoints were CD4 cell count below 200 x 10(6)/l, AIDS and death from AIDS. RESULTS: Sixty-three homo /bisexual men and 125 IDU fulfilled the entry criteria, with no significant differences in age at or date for seroconversion. Mean follow-up times were 6.7 and 7.0 years, respectively. The homo-/bisexual group had a significantly accelerated progression rate to all three endpoints: time to CD4 cell count below 200 x 10(6)/l (P = 0.002), to AIDS (P = 0.0003), and to death from AIDS (P < 0.0001). Adjusting for age and sex only made marginal alterations. Ten years after infection, 54% of homosexual men had developed an AIDS-defining condition and 51% had died from AIDS, whereas the corresponding precentages in the IDU group were 26 and 15, respectively. There was, however, no difference in overall mortality due to almost constant, non HIV-related, yearly mortality of some 4% in IDU. CONCLUSIONS: In our cohort there was a highly significant difference in disease progression and death from AIDS between homo-/bisexual men and IDU. This difference was proposed to be due to the transmission route determining the initial immune response and suggested that this route may have played a more important role than virus variability of the subsequent prognosis. PMID- 9223736 TI - Immunologic and virologic evaluation after influenza vaccination of HIV-1 infected patients. AB - OBJECTIVE: The present study was designed to determine the effect of immune activation, achieved by influenza vaccination, on plasma HIV RNA levels and immunological parameters including CD4 cell levels, antigen-stimulated T-cell function and apoptotic death of peripheral blood mononuclear cells. DESIGN AND METHODS: Thirty-four HIV-infected individuals and nine uninfected controls were immunized with influenza vaccine and blood was collected at weeks 0, 2, 4 and 16. Plasma was isolated and used for HIV RNA and influenza-specific antibody qualifications. CD4 cell counts, activation and maturation markers of T lymphocyte subsets were determined by flow cytometry. In vitro T-helper responses, spontaneous- and activation-induced cell death assays were also performed. RESULTS: Influenza-specific humoral and cellular immune responses correlated with CD4 count. Only in patients with CD4 counts > 300 x 10(6)/l there was a modest increase in T-cell responses to influenza virus, which was less than control subjects, observed after vaccination. Immunization had no significant effect on CD4 counts or plasma viral levels in the HIV-positive patients. Baseline apoptosis inversely correlated with CD4 counts and directly correlated with viral load. Activation-induced apoptosis did not change appreciably after vaccination and spontaneous apoptosis increased only in the < 300 CD4 group. CONCLUSION: These results indicate that immune stimulation resulting from influenza vaccination did not significantly change the levels of plasma virus, CD4 cell counts, or activation-induced apoptosis in HIV-infected individuals, although an increase in the T-cell response to influenza and spontaneous apoptosis was observed in the > 300 and < 300 CD4 groups, respectively. PMID- 9223738 TI - How many HIV infections cross the bisexual bridge? An estimate from the United States. AB - OBJECTIVE: Most heterosexual women with AIDS have been infected by male sex partners who acquired HIV via injecting drug use or sex with men. The contribution of bisexuality to heterosexual HIV however, has been poorly quantified. In this paper, we estimate the number of HIV infections that spread from the homosexual community to women who have sex with bisexual men. METHODS: We developed an HIV transmission model and assigned values to the model's parameters using data from a probability survey of US cities with a high risk of HIV. RESULTS: We estimated that these are about 400 HIV infections transmitted annually from HIV-infected bisexual men in high-risk cities to their female sex partners; two-thirds of these infections are transmitted to main female partners and one-third to casual partners. Uncertainties in the value of model parameters lead to variation in expected HIV infections mostly within the range 200 to 600, and for one parameter up to nearly 800. CONCLUSION: We conclude that transmission via bisexuality is a relatively minor component of the estimated 40,000 annual HIV infections in the USA. PMID- 9223737 TI - Trends in HIV-1 prevalence may not reflect trends in incidence in mature epidemics: data from the Rakai population-based cohort, Uganda. AB - OBJECTIVES: To assess whether trends in serial HIV-1 prevalence reflect trend in HIV incidence, and to decompose the effects of HIV-1 incidence, mortality, mobility and compliance on HIV-1 prevalence in a population-based cohort. DESIGN: Two-year follow up (1990-1992) of an open cohort of all adults aged 15-59 years, resident in a sample of 31 representative community clusters in rural Rakai District, Uganda. METHODS: A detailed household enumeration was concluded at baseline and in each subsequent year. All household residents were listed, and all deaths and in- and out-migrations that occurred in the intersurvey year wee recorded. In each year, all consenting adults were interviewed and provided a serological sample; 2591 adults aged 15-59 years were enrolled at baseline. RESULTS: HIV prevalence among adults declined significantly 1990 and 1992 (23.4% at baseline, 21.8% in 1991, 20.9% in 1992; P < 0.05). Declining prevalence was also observed in subgroups, including young adults aged 15-24 years (from 20.6 to 16.2% over 3 years; P < 0.02), women of reproductive age (from 27.1 to 23.5%; P < 0.05), and pregnant women (from 25.4 to 20.0%; not significant), However, HIV incidence did not change significantly among all adults aged 15-59 years (2.1 +/- 0.4 per 100 person-years of observation (PYO) in 1990-1991 and 2.0 +/- 0.3 per 100 PYO in 1991-1992], nor in population subgroups. HIV-related mortality was high (13.5 per 100 PYO among the HIV-positive), removing more infected persons that were added by seroconversion. Net out-migration also removed substantial numbers of HIV-positive individuals. CONCLUSIONS: In this mature HIV epidemic, HIV prevalence declined in the presence of stable and incidence. HIV-related mortality contributed most to the prevalence decline. Prevalence was not an adequate surrogate measure of incidence, limiting the utility or serial prevalence measures in assessing the dynamics of the HIV epidemic and in evaluating the impact of current preventive strategies. PMID- 9223739 TI - Assortative sexual mixing in a heterosexual clinic population--a limiting factor in HIV spread? AB - OBJECTIVE: To assess the degree of sexual mixing in a sexually transmitted disease clinic population stratified by country of birth. DESIGN: Prospective linked HIV serosurvey incorporating demographic and sexual risk data gathered by a doctor-administered questionnaire. SETTING: The Department of Genitourinary Medicine at St Thomas' Hospital, London, UK. SUBJECTS: Fifteen thousand eight hundred and seventy-eight heterosexuals who attended between April 1992 and February 1995. MAIN OUTCOME MEASURE: The degree of assortative (like-with-like) mixing, after stratification of the population by country of birth, of index patients, their parents and their sexual partners. RESULTS: Sexual mixing in this population of sexually transmitted disease clinic attenders is highly assortative when the CoB of parents (family origin) of index patients is taken into account. CONCLUSION: Our findings help to explain the low spread of heterosexual HIV infection in the UK to date, and may help future projections, and health targeting of those at risk. This model can be applied to other mixed population. PMID- 9223740 TI - On-site, rapid HIV testing with same-day results and counseling. AB - BACKGROUND: New rapid HIV antibody tests have allowed provision of results and result-specific counseling on the day on initial visit, and have the potential to increase the efficiency of HIV counseling and testing. METHODS: To evaluate the use of rapid testing with same-day results in public clinics, the Single Use Diagnostic System HIV-1 rapid assay was used for a 3-month period at an anonymous testing clinic and a sexually transmitted disease (STD) clinic in Dallas, Texas. Non-reactive rapid test results were reported as HIV-negative. Reactive results were reported as 'preliminary positive'. These procedures were compared with standard testing during a baseline period, with respect to number of clients receiving results and post-test counseling, client satisfaction, counselor acceptance, cost and effectiveness at reducing HIV risk. RESULTS: Rapid testing resulted in an increase in the number of persons learning their serostatus: a 4% increase for uninfected and a 16% increase for infected clients at the Anonymous Testing Clinic; a 210% increase for uninfected patients and a 23% increase for infected patients at the STD clinic. Rapid testing resulted in a cost saving of US$ 11 per test in both the anonymous and STD clinics. Of those previously tested, 88% responded that they preferred the rapid test. In the year following initial HIV test, clients tested with rapid and standard procedures were equally likely to return to the clinic with a new STD (odds ratio, 0.97; 95% confidence interval, 0.7-1.4). CONCLUSIONS: Rapid, on-site HIV testing was feasible, preferred by clients, and, resulted in significant improvement in the number of persons learning their serostatus, without increasing the costs or decreasing the effectiveness of counseling and testing. PMID- 9223741 TI - Does HIV-2 infection provide cross-protection against HIV-1 infection? PMID- 9223742 TI - Changes in plasma HIV RNA levels and CD4 cell counts after vaccination of pediatric patients. PMID- 9223743 TI - Quantification of HIV-1 RNA in cervicovaginal secretions: an improved method of sample collection. PMID- 9223744 TI - Detection of apoptotic cells from peripheral blood of HIV-infected individuals using a novel monoclonal antibody. PMID- 9223745 TI - The production of beta-chemokines induced by HIV-2 envelope glycoprotein. PMID- 9223747 TI - Condoms used during most commercial sex acts in Thailand. PMID- 9223746 TI - Baclofen for treatment of persistent hiccups in HIV-infected patients. PMID- 9223748 TI - Epidemiology of Mycobacterium avium infection in northern Italy. PMID- 9223749 TI - Oral methionine may improve neuropsychological function in patients with AIDS myelopathy: results of an open-label trial. PMID- 9223750 TI - Effects of indinavir treatment on platelet and neutrophil counts in patients with advanced HIV disease. PMID- 9223751 TI - Foscarnet-induced hypercalcaemia in AIDS. PMID- 9223752 TI - Role of trimethoprim-sulphamethoxazole in preventing HIV-associated bacteraemia. PMID- 9223753 TI - Gender difference in CD4+ cell counts persist after HIV-1 infection. SEROCO Study Group. PMID- 9223754 TI - Absence of detectable maternal DNA and identification of proviral HIV in the cord blood of two infants who became HIV-infected. PMID- 9223755 TI - Expression of three cell adhesion molecules in bladder carcinomas: correlation with pathological features. AB - Recently, independent studies have shown that the expression of two integrin chains, beta 4 and alpha 2, plus the epithelial cadherin are related to tumour progression in human bladder carcinomas. For the first time, we compare the expression of these three cell adhesion molecules using immunohistochemical analysis of consecutive cryosections from a series of 50 bladder tumors. E cadherin, beta 4, and alpha 2 were strongly expressed in normal urothelium. A majority of non-invasive bladder cancers stained positively for E-cadherin (62%), whereas only 29% expressed normal positivity for alpha 2, and only 35% for beta 4. However, most invasive tumours presented an aberrant expression of alpha 2 (81%), beta 4 (100%), and E-cadherin (75%). We studied the correlation of immunoreactivity with histological grade and stage. The alpha 2 pattern was not correlated with stage and grade. In contrast, loss of normal beta 4 expression was significantly related to increasing tumour grade and deep invasion with a higher correlation for grade. Finally, E-cadherin expression was highly correlated with stage, but not with grade. Thus our results indicate that, although many invasive bladder tumours presented a disorder in expression of the two integrins alpha 2 and beta 4, E-cadherin appeared to be a better market of invasiveness in bladder carcinomas. PMID- 9223756 TI - Signal amplification of FISH for automated detection using image cytometry. AB - The purpose of this study was to improve the detection of FISH signals, in order that spot counting by a fully automated image cytometer be comparable to that obtained visually under the microscope. Two systems of spot scoring, visual and automated counting, were investigated in parallel on stimulated human lymphocytes with FISH using a biotinylated centromeric probe for chromosome 3. Signal characteristics were first analyzed on images recorded with a coupled charge device (CCD) camera. Number of spots per nucleus were scored visually on these recorded images versus automatically with a DISCOVERY image analyzer. Several fluochromes, amplification and pretreatments were tested. Our results for both visual and automated scoring show that the tyramide amplification system (TSA) gives the best amplification of signal if pepsin treatment is applied prior to FISH. Accuracy of the automated scoring, however, remained low (58% of nuclei containing two spots) compared to the visual scoring because of the high intranuclear variation between FISH spots. PMID- 9223757 TI - Trends in registered psychiatric morbidity and forecasting psychotropic drug needs in Bulgarian hospitals. AB - Registered psychiatric morbidity in Bulgaria as a whole and particularly in Plovdiv, the second largest region of the country, was assessed. Three aspects of psychotropic drug usage were analysed, namely, changes in registered psychotropic drugs, the prescribed daily dose (PDD) values for 2 years, and the preferred therapeutic schemes, and drug usage and needs in a psychiatric hospital with 365 beds. This was done by time series analysis for evaluation of psychiatric morbidity and drug consumption data, calculation of PDD for psychotropic medicines, and, based on a modification of the World Health Organization's morbidity method, assessment and prediction of drug use and needs in Plovdiv hospital. The results indicated that the registered morbidity had increased by 4% over the period 1989-93 to 2,427 psychiatric patients per 100,000 people. The increased consumption of especially benzodiazepines and sedative medicines was analyzed. Diazepam was prescribed the most often (91.1%), followed by levomepromazine (86.4%), haloperidol (82.7%), etc. Future drug consumption in Plovdiv hospital is expected to decrease because therapeutic practice in hospitals has been revised and improved on the basis of the World Health Organization's recommendations. PMID- 9223758 TI - Preparation and characterization of a new polymorphic form and a solvate of glibenclamide. AB - A new crystalline form and a solvate of glibenclamide were prepared and characterized by hot-stage microscopy, DSC, IR, scanning electron microscope, X ray powder diffraction and dissolution studies. From I, III and the solvate which melt at 174.4, 153.2 and 109.3 degrees C, were obtained by crystallization. Significant differences between the three forms in the X-ray and IR spectra were observed. Polymorphic transformation was observed for the least stable form (III) to the most stable one (form I) upon storage at different temperatures and trituration. The dissolution profiles showed significant differences between the three forms, with form III having the highest dissolution rate. PMID- 9223759 TI - Simultaneous determination of the binary mixture of nifedipine and mefruside using derivate spectroscopy, capillary gas-liquid chromatography and high performance liquid chromatography. AB - Accurate, precise first-derivative ultraviolet spectrophotometric, gas-liquid and high performance liquid chromatographic methods for the determination of nifedipine and mefruside in tablet dosage form were described. The first derivative method depends on measurements of the derivative amplitudes by peak-to base and zero-crossing techniques, at 390 and 282 nm, for the determination of nifedipine and mefruside, respectively. Calibration graphs were linear (r = 0.9999) ranging from 8-40 and 20-60 micrograms ml-1 for nifedipine and mefruside, respectively, in presence of one another. The gas-liquid chromatographic method (GLC) was based on using cross-linked methylsilicone gum capillary column with flame ionization detector for the determination of nifedipine and mefruside in the binary mixture. The high performance liquid chromatographic (HPLC) method was based on using a reverse-phase column with a mobile phase of methanol-water (55 45, pH = 4.5) with UV detection at 260 nm. The three methods showed good linearity, precision and reproducibility. The proposed methods were successfully applied to the determination of both drugs in pharmaceutical tablets. PMID- 9223761 TI - Health promotion attitudes, practices and obstacles of Italian physicians with their patients. PMID- 9223760 TI - Synthesis and evaluation of some acyloxyethyl mefenamates as possible prodrugs. AB - Various acyloxyethyl mefanamates were synthesized and evaluated for potential application as prodrugs. Their kinetics of hydrolysis were examined in aqueous solutions of pH 1.0 and 7.4 and in human plasma at 37 degrees C. Among the synthesized compounds, the beta-carboxypropionylethyl mefenamate and the pivaloyloxyethyl mefenamate show high stability against enzymatic and non enzymatic hydrolysis. On the other hand the acetyloxyethyl mefenamate shows t1/2s of 1.4 h, 1.41 h and 3.61 h in human plasma, solutions of pH 7.4 and pH 1.0 respectively; However, its hydrolysis to mefenamic acid in plasma was not quantitative. Preliminary in vivo study shows that acetyloxyethyl mefenamate gave plasma concentration of mefenamic acid lower than that of control after oral administration. The calculated AUC0-inf for the acetyloxyethyl and control were 45 and 85 micrograms.h/ml respectively. PMID- 9223763 TI - Pitfalls in managing routine medical problems of patients with spinal cord injury. PMID- 9223762 TI - Lipid-lowering therapy after coronary artery bypass surgery: the Post-CABG trial. AB - The Post Coronary Artery Bypass Graft trial is an important milestone that documents the benefits of treating hypercholesterolemia in patients with severe coronary atherosclerosis who have had bypass surgery. In the present article, we highlight the rationale, study design, and results of the Post-CABG study, which was recently published in the New England Journal of Medicine. PMID- 9223764 TI - How physicians can prevent medication errors: practical strategies. AB - To eliminate and reduce medication errors, health care organizations must develop a consistent approach that allows examination of errors in a supportive atmosphere with a bias toward preventing future errors rather than punishing past ones. Until improved systems are in place, physicians can help prevent many of the most serious medication errors by observing some basic safety practices, such as writing orders whenever possible and limiting verbal orders to urgent or emergency situations, writing clearly and neatly, and avoiding abbreviations. PMID- 9223765 TI - Arthralgias, myalgias, facial erythema, and a positive ANA: not necessarily SLE. PMID- 9223766 TI - Adult respiratory distress syndrome (ARDS): current management, future directions. AB - The adult respiratory distress syndrome, marked by severe, refractory hypoxemia, noncardiogenic pulmonary edema, and stiff, noncompliant lungs, demands quick recognition and intensive care. This article reviews the disease process and current and experimental treatments for it. PMID- 9223767 TI - Standards of care for treating headache in primary care practice. National Headache Foundation. AB - The following is a summary of guidelines created under the auspices of the National Headache Foundation, in an effort to improve the care of headache patients in primary care practice. The guidelines represent the consensus of an advisory panel of practitioners chosen by the NHF for their expertise in four specialty areas. A complete set of guidelines can be obtained by calling the National Headache Foundation at 1-800-843-2256 or by writing to them at 428 W. St. James Pl., 2nd Floor, Chicago, IL 60614-2750; the cost is $10. PMID- 9223769 TI - New guidance on post exposure prophylaxis of health care workers exposed to HIV at work. PMID- 9223768 TI - Inpatient management of acute leukemia. AB - Inpatients with acute leukemia need meticulous supportive care, which can be complex and challenging. The physician must be vigilant for infection, hemorrhage and other complications. Aggressive transfusion support is often necessary, but can itself cause complications. PMID- 9223770 TI - Foreword: the key role of research in making rational mental health services. PMID- 9223771 TI - Population needs for mental health services. Introduction to the theme. PMID- 9223772 TI - The Mental Illness Needs Index. PMID- 9223773 TI - Assessment of needs for care among patients with schizophrenic disorders 15 and 17 years after first onset of psychosis. PMID- 9223774 TI - Do we still need mental hospitals? PMID- 9223775 TI - How many psychiatric beds: towards a needs based portfolio of residential care. PMID- 9223776 TI - Population needs assessment for mental health services. Prospects for the future. PMID- 9223777 TI - Describing mental health services: the development of a mental health census in the north-west of England. PMID- 9223778 TI - Computers and process description for community mental health care. PMID- 9223779 TI - Mental health services description. Prospects for the future. PMID- 9223780 TI - Service utilization: a pivotal measure in assessing service needs and service outcome. PMID- 9223782 TI - Service utilization research: goals and prospects. PMID- 9223781 TI - Social networks and service utilisation in patients with severe mental illness. PMID- 9223783 TI - Importance of local differences in comparing hospital and community psychiatric services. PMID- 9223784 TI - Partial or full-time hospitalization: patients' preference. PMID- 9223785 TI - Economics and mental health: a concise European history of demand and supply. PMID- 9223786 TI - Costing day hospital and in patient care for acute psychiatric illness. PMID- 9223787 TI - Psychiatric case registers for monitoring service utilisation and evaluating its costs. PMID- 9223788 TI - The cost of mental health services. Prospects for future research. PMID- 9223789 TI - Quality development in mental health care in Europe. Recent contributions by WHO. PMID- 9223790 TI - Community psychiatry in Europe: assessment and evaluation. PMID- 9223791 TI - The importance of service level measures for mental health policy. PMID- 9223792 TI - Evaluating mental health services. A world perspective. PMID- 9223793 TI - [Liver allografts from donors older than 60: benefits and risks]. AB - With limited organ resources and an increasing number of candidates for liver transplantation, the world-wide trend is towards using liver allografts from donors older than 60 years. This strategy, however, may be hazardous because of the known correlation between advanced donor age and graft dysfunction. Since January 1996, each of 5 patients received a liver allograft from a donor older than 60 years. Preservation time in these cases was shortened as much as possible and liver allografts were used only if there were no other potential risk factors for primary nonfunction. Mean cold ischemic time was significantly shorter in this donor group (7.8 hrs) than for livers from 28 younger donors (10.2 hour; p < 0.01). 3 of the 5 grafts from older donors had normal function immediately. The other 2 initially had biochemical features of preservation injury, but graft function returned to normal within the first week after transplantation. All 5 patients currently have normal graft function, with follow-up ranging from 3-8 months. There was no difference between the 5 recipients of grafts from older donors and 28 adult recipients of grafts from younger donors in extent of preservation injury and in immediate graft function. We conclude that in countries with limited organ resources, such as Israel, liver allografts from older donors can be used within defined limits and minimal preservation time. PMID- 9223794 TI - [High dose oral prednisone for hemangiomas in infants]. AB - Over a 24-year period, 62 infants (47 girls) with hemangiomas were treated with an initial dose of either 3 or 5 mg/kg/day of oral prednisone for 2 weeks, after which the dose was gradually tapered off during 6-8 weeks. Few patients required longer treatment. Results were judged to be excellent in 68% of infants and good in 25%. Treatment was considered a failure in only 7%. The initial dose of 5 mg/kg/day was more effective than the smaller dose (p < 0.001). Of the 62 patients, 49 received 1 course of treatment, 8 required 2 courses and 5 required 3 courses. Retreatment was given whenever significant regrowth occurred. Side effects were not serious, and resolved when treatment was discontinued. Treatment was indicated when the location of the lesions caused interference with important functions or when the lesions were likely to damage anatomic structures. Special attention was paid to early treatment of eye and subglottic hemangiomas. In all 22 children with hemangiomas of the eye (most with an orbital component), shrinkage of the lesion was observed within 24 hours of initiating treatment. In 19 of the 22 there was no residual of the hemangioma 1-18 years later. Such lesions deserve early treatment, not just as cosmetic emergencies, but to prevent secondary amblyopia. Early treatment of subglottic hemangiomas is also mandatory because they are potentially life-threatening. We conclude that oral prednisone is very effective in the treatment of hemangiomas of infants when given at a high dose for an adequate period of time. PMID- 9223795 TI - [Angina pectoris and the severity of coronary artery stenosis]. AB - The relationship between angina pectoris and the severity of coronary artery disease was evaluated in 146 patients with normal segmental and global, left ventricular, systolic performance. None had unstable angina or a previous myocardial infarction. A strong relationship was found between angina and the severity of coronary artery disease (p < 0.005). Significant, stable, angina pectoris as a clinical symptom indicated advanced coronary artery disease in this selected group of patients. PMID- 9223796 TI - [Salivary gland endoscopy: a new technique for diagnosis and treatment of sialolithiasis]. AB - The use of an endoscopic, minimally invasive technique for the removal of salivary gland stones from the submandibular or parotid duct is described. A 2.0 2.7 mm endoscope is inserted into an incision in the parotid or submandibular duct. When the stone is visualized through the endoscope it is removed using suction and forceps. We used this technique in 45 cases for removal of calculi, screening the ductal system to rule out residual calculi and determination of ductal dilatation. The success rate was 80% and there were no major postoperative complications. To the best of our knowledge these are the first such cases reported in Israel. PMID- 9223798 TI - [Open access upper endoscopy--"good or bad for the Jews?"]. PMID- 9223799 TI - [Molecular genetics of Wilm's tumor]. PMID- 9223797 TI - [ATLS course in emergency medicine for physicians?]. AB - Implementation of Advanced Trauma Life Support (ATLS) skills among practicing physicians and its perceived utility in their civilian practices, as well as in their potential army combat assignments, was evaluated. 177 physicians in various subspecialties, who were graduates of ATLS training courses, answered a specially designed telephone questionnaire. An unexpectedly high percentage of physicians (47%) had used their ATLS training when called to treat trauma victims. 67% of physicians stressed the contribution of the ATLS course to enhancing their skills. We believe that a properly designed ATLS course for general practitioners would be very beneficial for trauma victims. PMID- 9223800 TI - [Sentinel node sampling in patients with malignant melanoma]. PMID- 9223801 TI - [Malignancy in Crohn's disease]. PMID- 9223802 TI - [Administration of fluid by subcutaneous infusion: revival of a forgotten method]. PMID- 9223803 TI - [Libman-Sacks endocarditis--a jubilee year]. PMID- 9223804 TI - [Various aspects of circumcision]. PMID- 9223806 TI - [Awareness of ovulation time and sex determination in Biblical times]. PMID- 9223805 TI - [Liver dysfunction induced by Epstein-Barr virus infectious mononucleosis]. PMID- 9223807 TI - [Biological fixation of distal tibial fractures]. PMID- 9223808 TI - [Continuing medical education]. PMID- 9223809 TI - [Rabies--re-emerging threat?]. PMID- 9223810 TI - [Dr. Max Nordau--physician, writer, journalist, and Zionist leader]. PMID- 9223811 TI - [Effect of the Yom Kippur fast on parturition]. AB - Food-withdrawal has been proposed as a possible mechanism for initiating the onset of labor in animals and humans. The hypothesis was based upon the reported increase in deliveries of infants during the Yom Kippur fast. We studied the effect of the fast on full term deliveries of Jewish women, with non-fasting Bedouin women as controls (1988-1995, 1,313 Jewish and 1,091 Bedouin deliveries). To determine the effect of Yom Kippur itself, delivery rates on Sukkot and Yom Kippur were compared in both groups. The mean delivery rate in the Jewish population was significantly higher during Yom Kippur and the day after, than during the 7 days before Yom Kippur (15.1 +/- 5.1 and 14.6 +/- 4.7 vs 10.7 +/- 3.5, p < 0.04 and p < 0.01, respectively). There was an increase in delivery rate during the 6 hours before the end of the fast. In the Bedouin women there were no changes in delivery rate during any of these periods. There were no significant differences in the rates of deliveries during the Sukkot festival between Jewish and Bedouin women. We conclude that fasting is associated with a significant increase in the rate of deliveries at term. PMID- 9223812 TI - [Pulmonary alveolar microlithiasis presenting with prolonged cough]. AB - A 40-year-old man had been followed in the pulmonary clinic for prolonged cough. Chest X-ray showed bilateral diffuse interstitial infiltrates with accentuation toward the bases. CT-scan demonstrated a fine diffuse reticulonodular pattern. Transbronchial lung biopsy showed pulmonary alveolar microlithiasis, a rare disease characterized by the presence of concentric calcifications within the pulmonary alveoli. This is the second case of the disease reported in Israel. PMID- 9223813 TI - [Laser photorefractive surgery]. AB - As photorefractive keratectomy (PRK) with excimer laser gains world-wide acceptance, more patients are able to discard their spectacles. Our study comprised 611 eyes which underwent PRK and were followed for at least a year. Those with myopia up to -6.00D had better post-operative visual acuity and refraction, than patients with higher grades of myopia. They had less corneal haze and a greater proportion were satisfied with their results. Complaints of halos and glare were similar at all degrees of myopia up to -10.00D. PMID- 9223814 TI - [Oral anticoagulation therapy in the primary care setting]. AB - The use of oral anticoagulant therapy (OAT) to prevent thromboembolism has been widespread in recent years. The concept of high- and low-intensity regimens has facilitated treatment for many, and has lowered the hazards of overly intense anticoagulation. However, a significant proportion of patients suited to the low intensity regimen are not being treated. It is not clear whether its wider use is limited by continued debate, lack of resources, lack of expertise, or other causes. We retrospectively reviewed the medical records of 32 patients treated with OAT administered in the primary care setting. The average age was 66 +/- 11 years (range 34-84). 9 were treated with high-intensity OAT: 8 due to artificial heart valves, and 1 due to a hypercoagulable syndrome with recurrent thromboembolism. 23 were treated with low-intensity OAT, 17 of whom had atrial fibrillation. 11 were also being treated continuously with other medication which interacted with OAT or interfered with other coagulation pathways. Such medication included: aspirin, dipyridamole, amiodarone, bezafibrate and allopurinol. Of 414 coagulation tests, 57% and 65% were in the therapeutic range in the high- and low-intensity OAT groups, respectively. There was no major bleeding event, but in 2 of 8 who bled, gastrointestinal bleeding led to hospitalization. Treatment was discontinued in 1 patient because of difficulties in achieving target INR, and in the 2 hospitalized for bleeding. The percentages of test results in, above and below the therapeutic range were similar to those in other large series, for both intensity regimens. We found that a significant proportion of patients were under chronic treatment with other medication which interacted with OAT. To estimate the rate of complications in primary care OAT, larger series are needed. We conclude that OAT can be given and monitored by the family physician, and that awareness of long and short term drug interactions with OAT is mandatory. PMID- 9223816 TI - [Chronic tophaceous gout]. AB - A 51-year-old immigrant from the Caucasus had had chronic tophaceous gout for over 20 years, but had never been treated with anti-hyperuricemic drugs. He had developed large, multiple tophi in many locations, including both ankles and feet. The enormous size and unique location of the tophi caused considerable pain, and difficulty in standing and on walking. Since surgical removal of the tophi was refused by the patient, a course of allopurinol, 300 mg/day, was begun. PMID- 9223815 TI - [Inhaled budesonide for chronic obstructive pulmonary disease]. AB - A significant, large minority of patients with chronic obstructive pulmonary disease (COPD) respond favorably to corticosteroid treatment; but the benefit may be outweighed by its side effects. Long-term administration of inhaled steroids is a safe means of treatment. We hypothesized that treatment with inhaled budesonide would improve clinical symptoms and pulmonary function in subjects with COPD, and that the response to an inhaled B2-agonist would individualize steroid responders. In 44 patients with stable COPD in a double- blind crossover trial, we compared a 6-week course of inhalations of 800 micrograms/d budesonide with a placebo, separated by a 4-week interval when no medication was taken. In 33 out of 42 responders to the B2-agonist who remained in the study, there was a significant improvement in FEV1 of greater than 20% following budesonide inhalation, as compared to placebo. There was also a significant difference between the 2 periods of treatment as to the mean number of B2-agonist inhalations. We conclude that about 1/4 of patients with stable COPD respond to bronchodilators, and treatment with inhaled steroids improves spirometry data and inhaled B2-agonist consumption in about 3/4. PMID- 9223817 TI - [The value of clinical information in medical imaging]. PMID- 9223818 TI - [Acute pancreatitis in childhood--new perspectives]. PMID- 9223819 TI - [Cerebral salt wasting syndrome]. PMID- 9223820 TI - [Congenital adrenal hyperplasia--is there a need for a national screening program]. PMID- 9223821 TI - [Evaluation and treatment of swallowing disorders (dysphagia)--the American model as an opportunity for improving patient care and cost containment in Israel's healthcare system]. PMID- 9223822 TI - [Epilepsy and higher mental functions]. PMID- 9223823 TI - [Bacterial translocation from the intestinal tract--the phenomenon and its clinical significance]. PMID- 9223824 TI - [Symptomatic management of children with the common cold]. PMID- 9223825 TI - [The use of prophylactic antibiotics in common infections in the community]. PMID- 9223826 TI - [99m technetium-dimercaptosuccinic acid renal scan in urinary tract infection in infancy and childhood]. PMID- 9223827 TI - [Surgical treatment of inguinal hernia in infants and children]. PMID- 9223828 TI - [Testicular varicocele in adolescents]. PMID- 9223829 TI - [Cholera vaccines]. PMID- 9223830 TI - [Immediate orthopedic surgery in multiple trauma]. AB - The increase in mortality and morbidity from multiple trauma due to road accidents, industrial trauma and combat injuries obligates treatment based on emergency systems and trauma centers. Follow-up of the frequency of different types of injury calls attention to increasing involvement of the orthopedic surgeon in primary treatment. This situation calls for appropriate deployment of immediate surgical treatment which will rapidly enable mobility. We present several methods for immediate orthopedic treatment of multiple-trauma patients. PMID- 9223831 TI - [Prioritizing the initial management of the multiple injured patient]. PMID- 9223832 TI - Use of herbal preparations for intractable cough. PMID- 9223833 TI - Re: The ethics of death-hastening or death-causing palliative analgesic administration. PMID- 9223834 TI - Re: The ethics of death-hastening or death-causing palliative analgesic administration. PMID- 9223836 TI - Hydration for control of syncope in palliative care. PMID- 9223835 TI - Acute myofascial pain after regional anesthesia. PMID- 9223837 TI - Long-term efficacy and safety of dronabinol for acquired immunodeficiency syndrome-associated anorexia. AB - We studied the effects of long-term (12 months) dronabinol in 94 late-stage acquired immunodeficiency syndrome (AIDS) patients (mean CD4 count of 45/mm3) who previously participated in a 6-week study (placebo versus dronabinol). All patients received dronabinol orally-2.5 mg twice daily (90%) or 2.5 mg once daily (10%). Appetite was measured using a visual analogue scale for hunger (VASH). Dronabinol was associated with consistent improvement in mean appetite. Patients previously treated with dronabinol continued to show improvement in VASH (percent change from baseline of 6-week trial: 48.6-76.1% at each month), whereas those previously treated with placebo exhibited substantial improvement in mean appetite, particularly during the initial 4 months of treatment (48.5-69.9%). Thereafter, dronabinol was associated with a VASH change at least twice baseline. Patients tended toward stable body weight for at least 7 months. Adverse events were primarily related to known central nervous system effects of dronabinol. These data support long-term, safe use of dronabinol for anorexia associated with weight loss in patients with AIDS. PMID- 9223838 TI - Opioid-sparing effect of diclofenac in cancer pain. AB - This study investigated the opioid-sparing effect of diclofenac using patient controlled analgesia with oral methadone. Fifteen patients with advanced cancer participated. After achieving adequate analgesia with regular dosing of oral methadone (T1), patient-controlled analgesia with methadone was administered for 3 days (T2). Intramuscular diclofenac 75 mg twice daily was then added to this regimen for 3 days (T3). Compared to T2 values, methadone dose was significantly reduced at T2 and T2, and pain report (recorded on a visual analogue scale) was significantly reduced at T3. A reduction in methadone plasma concentration was also observed at T2 and T3, although it did not attain statistical significance. Significant decreases in the intensity of several symptoms other than pain were also found at T2 and T3. Diclofenac appears to have a relevant opioid-sparing effect when using patient-controlled analgesia with oral methadone. PMID- 9223839 TI - Assessment of knowledge about cancer pain management by physicians in training. AB - This survey assessed the knowledge of physicians in training about the pharmacology of opioid analgesics and the benefits of palliative radiation therapy in the management of cancer pain. Eighty-one trainees at the Washington University Medical Center completed a questionnaire that addressed the palliative care of a hypothetical patient with metastatic non-small cell lung cancer. The questions addressed were 1) opioid selection, 2) conversion of parenteral to oral morphine, 3) management of opioid toxicities, 4) opioid addiction, and 5) efficacy of radiation therapy. The results demonstrated that few physicians in training were familiar with the stepwise progression of analgesic selection outlined in the World Health Organization (WHO) guidelines. When asked to convert a parenteral dose of morphine to an equivalent dose of a controlled-release preparation, 75% calculated a dose that was less than one-third the correct dose; only four (5%) calculated the dose correctly. Trainees were familiar with the management of opioid toxicities. They were unfamiliar with the palliative benefits of radiation therapy. Although 41% recognized that complete relief of pain could be achieved in 50%-60% of patients, most (70%) predicted that maximum pain relief would be seen within the first month, and 98% predicted maximum benefit by 12 weeks. Although cancer pain management has been highlighted in the lay and medical literature, physicians in training still demonstrate deficiencies in their knowledge about the pharmacology and bioequivalency of the opioid and the benefits of radiation therapy. Published guidelines for the management of cancer pain need to be disseminated to all medical personnel caring for patients with cancer. PMID- 9223840 TI - A multicenter evaluation of cancer pain control by palliative care teams. AB - Data on pain prevalence and severity were collected prospectively from advanced cancer patients as an integral part of two service evaluations. Six multidisciplinary palliative care teams working in Ireland formed the basis of one study and five teams based in the South of England were included in the second. A total of 695 cancer patients were referred and died in care in a minimum 6-month data collection period. Of these, 70% (486/695) were experiencing pain at referral to the services. After 2 weeks, there was a significant reduction (P < 0.0001) in the levels of pain experienced by patients, and no patient had overwhelming pain. The data emphasize that pain prevalence in advanced cancer patients cared for in the community is as high as that observed in other settings. Multidisciplinary palliative care teams are shown here to be effective in alleviating pain. PMID- 9223841 TI - Monitoring practices following epidural analgesics for pain management: a follow up survey. AB - Indwelling epidural catheters are commonly used in pain management. There is much literature on the risks and benefits of this therapy, but there is no consensus on a standard, cost-effective method of monitoring these patients, nor on the types of analgesics or methods of drug delivery. The purpose of this paper is to examine clinical practices of pain management nurses belonging to the American Society of Pain Management Nurses (ASPMN). A total of 202 members responded to a follow-up survey on their institutions' use of epidural catheters. Although there were numerous variations in practice, certain similarities and themes regarding drug choice, patient monitoring, and health care providers involved with the care of this patient population are evident. PMID- 9223842 TI - Cancer pain emergencies: a protocol for management. AB - Cancer is often associated with chronic pain, which can be managed by established algorithms. Cancer is also occasionally associated with sustained episodes of excruciating pain (cancer pain emergencies) that require rapid application of powerful analgesic strategies in a manner that is distinct from chronic pain management techniques. To clarify the utility of rapid opioid dose escalation in this setting, we reviewed the management of ten cancer pain emergencies in nine patients. After initial assessment, all patients were managed according to a protocol whereby intravenous boluses of opioid are administered with rapid upward titration until an effective analgesic dose is found. Using this technique, all patients had relief of their excruciating pain after a mean of 89 min (range, 4 215 min). No patient demonstrated evidence of significant toxicity. We conclude that repeated intravenous boluses of an opioid, doubling the dose every 30 min until analgesia is achieved, is effective and safe management of cancer pain emergencies. Validation of this protocol is required. PMID- 9223844 TI - Tuberculosis: a spectre has returned. PMID- 9223843 TI - Strychnine-like multifocal myoclonus and seizures in extremely high-dose opioid administration: treatment strategies. AB - While occasional myoclonic jerks are prevalent in cancer patients receiving opioids, severe myoclonic jerks and seizures due to opioids are uncommon. In this retrospective case series, we describe five cancer patients with refractory cancer pain and severe neuroexcitatory toxicity associated with extremely high dose opioid therapy to characterize better the syndrome, its treatment, and its outcome. Two patients died following seizures, but three patients recovered following prompt treatment with parenteral midazolam infusions and rotation to alternative opioids. Possible mechanisms and treatment options for this potentially lethal clinical syndrome are reviewed. The authors conclude that severe multifocal myoclonus and seizures associated with extremely high-dose opioid therapy are life-threatening, and respond to parenteral midazolam infusion, rotation to alternative opioids, and aggressive supportive care. PMID- 9223845 TI - A need to reduce the radon gas hazard in the UK. PMID- 9223846 TI - Work-related stress: implications for the employer. AB - This paper considers many of the aspects of work-related stress, including problems of definition, the legal duties and responsibilities of employers towards their employees concerning stress. It includes an outline of some of the possible financial consequences (and legal penalties) if employers do not, so far as is reasonably practicable, take appropriate action to remove, reduce or alleviate work-related stressors affecting their employees' health. The paper concludes with a summary of a range of coping strategies which will assist employers to comply with legislative requirements and duties. PMID- 9223847 TI - Knowledge and attitudes to HIV and AIDS and sexual practices among university students in Lusaka, Zambia and London, England: are they so different? PMID- 9223848 TI - Female circumcision and its health implications: a study of the Uruan Local Government Area of Akwa Ibom State, Nigeria. AB - A total of 400 subjects was randomly selected from 40 villages in the Uruan Local Government Area of Akwa Ibom State for the study. The purposes of the study were to: i. identify the 'established benefits' of female circumcision; ii. identify the health hazards that accompany the practice; and iii. create awareness among community members of the ill-effects of the practice. The study discovered a strong belief in the established benefits and poor appreciation of the health hazards of female circumcision by the participants. Recommendations were made for more efforts in public health education programmes on the ill-effects of the practice. Studies were also recommended to be conducted in other parts of the country to assess the level of awareness on the ill-effects of such an operation and the institution of educational programmes where applicable. PMID- 9223850 TI - Psychosocial assessment methods in childhood epilepsy: focus on sub-Saharan Africa. AB - The scale of evidence tilts towards the contention that epileptic children as a group are at a disadvantage regarding intellectual and emotional development and have higher rates of behavioural abnormalities than their non-epileptic peers. Differentiating factors logically point to the problem areas which should be enquired into in a comprehensive assessment of a child with epilepsy and should facilitate formulation of viable intervention stratagems. A critical review of the literature reveals that, of five major assessment methods, the use of a standardised and valid questionnaire is likely to yield the most reliable clinical information. However, a complete assessment package should include drug use monitoring, evaluation of cultural milieu and family psychodynamics. PMID- 9223849 TI - Infant mortality in the rural Riyadh region of Saudi Arabia. AB - A prospective study of a cohort of infants born in 1987 was carried out until they were one year old. Five villages were selected at random. All the babies born in 1987 in these villages were identified by a group of trained nurses. These nurses collected morbidity and mortality data on these children each time the events occurred using a structured data collection form. Data were analysed using relevant demographic and statistical techniques. A total of 4,963 babies was born during the period of study. The neonatal mortality rate was 21.4 per 1,000 live births and the infant mortality rate was 53.8 per 1,000. The postneonatal death rate was 32.5 per 1,000 live births. The causes of infant deaths as presumed from reported signs of last illness were gastroenteritis, respiratory problems, preterm birth complications and congenital abnormalities. It was concluded that there is a decline in the infant mortality rate compared with previous estimated rates, but the high levels of neonatal death rates call for improved antenatal and obstetric health services. The high postneonatal death rate indicates the potential for a further reduction. PMID- 9223851 TI - Social marketing: a tool not a solution. AB - There is a longstanding debate on the contribution of social marketing to public health in general, and to health education and health promotion in particular. This paper presents further discussion from a public health point of view and concludes that priority should be given to health-oriented approaches rather than market-oriented strategies. It is argued that, at best, social marketing is a tool not a solution for health education's and health promotion's problems. To communicate health education messages effectively and efficiently, health needs assessment is recommended as a way forward. It is a public health approach and contains a range of flexible methods in the implementation of health education/promotion programmes. PMID- 9223852 TI - Tuberculosis and the cow. PMID- 9223853 TI - Occupational disease and injury and state compensation. PMID- 9223854 TI - [Redo operation after surgery for aortic aneurysm and dissection]. AB - Thirteen patients who underwent redo operation after surgical treatment of aortic aneurysm and dissection were presented. In 8 patients, redo operations were performed for aortic dissection following aortic valve replacement. A-C bypass, the Koster-Collins operation and replacement of thoracic aorta. In the other 5 patients, the reasons for redo operation were aortic root enlargement after replacement of ascending aorta and aortic valve replacement, pseudoaneurysm and aneurysmal dilatation around coronary button for the Bentall operation and recurrent aneurysm after patch aortoplasty and thoracoabdominal replacement using the Crawford's maneuver. To prevent these redo operation, adequate selection of surgical procedures and meticulous operative techniques should be required in primary operation. PMID- 9223855 TI - [A case of thyroid cancer invading into mediastinum that was in need of resection of both innominate veins for complete cure]. AB - This case report describes a 59-year-old female patient with superior vena cava syndrome due to large thyroid cancer involving both innominate veins and left common carotid artery. The tumor was resected together with the both innominate veins and left common carotid artery. Right innominate veins and left common carotid artery were reconstructed with FEP-Ringed grafts. Microscopical finding was papillary carcinoma and its lymph node metastasis. Postoperative course was uneventful relatively. Three years after operation the patient is good health without any sign of recurrence. PMID- 9223856 TI - [Preoperative evaluation of the right ventricular function using the pulsed Doppler echocardiogram in the patients scheduled for the elective pulmonary resection]. AB - Preoperative evaluation of the both right ventricular (RV) systolic and diastolic function using the pulsed Doppler echocardiogram were examined for 46 patients scheduled for the pulmonary resection of the pulmonary tumorous lesions. The parameters which included the RV inflow pattern at the tricuspid orifice and the RV ejection flow pattern at the RV outflow tract were obtained by the pulsed Doppler echocardiogram. Following results were obtained. 1) The RV afterload shown by the parameters of the acceleration time (AT, time beginning of RV ejection to peak velocity) were higher in the aged patients, the low FEV 1% populations, and the patients having the deteriorated left ventricular contraction. 2) The RV diastolic dysfunction were present preoperatively in aged patients over the 60 years old. This phenomenon was characterized by a high degree of atrial contraction and an increased ratio of the peak velocity in atrial contraction phase to that in early rapid filling phase (A/E). When we evaluate the RV inflow and ejection Doppler flow patterns after the major lung resection, these findings must be considerable thing. PMID- 9223857 TI - [Replacement of the aortic root with woven and knitted Dacron composite graft]. AB - Some of the patients with annuloaortic ectasia or Stanford type A require aortic root replacement. If conventional Bentall procedure is employed in such cases, aortic root will become a cylindrical shape totally lacking sinus of Valsalva because the procedure utilizes a straight tube. The sinuses of Valsalva support an important role in opening and closing the valvar leaflets, and opening of the coronary arteries, together with the interleaflet triangles and the sinutubular junction. In this study, we performed aortic root replacement with composite graft which consisted of two types of graft including 30 mm Knitted Dacron Graft (Gelsoft) with different dilation rates and 30 mm Woven Dacron Graft (Hemashield) as well as bioprosthesis (27 mm Carpentier-Edwards), in order to reconstruct sinus of Valsalva. Post-operative angiography revealed an excellent diastolic coronary flow, as evidenced by proximal Knitted graft of 37 mm in diameter, distal Woven graft of 30.3 mm in diameter and Doppler flow DSVR (Diastolic/Systolic Velocity Ration) of 2.2 measured at the left coronary orifice. Since it is difficult to obtain homograft at present, this procedure would be worth trying during aortic root replacement. PMID- 9223858 TI - [Surgical treatment of infective endocarditis associated with cerebral mycotic aneurysm]. AB - We experienced two cases of infective endocarditis associated with cerebral mycotic aneurysm. Case 1: 58 year-old man underwent emergency aortic and mitral valve replacement due to active infective endocarditis and congestive heart failure diagnosed by transesophageal echocardiography. After the operation, he did not wake up and his bilateral pupils were dilated. Computed tomography demonstrated massive intracranial hemorrhage and severe brain edema. He died from multiple organ failure 22th postoperative day. Rupture of cerebral mycotic aneurysm was strongly suspected. Case 2: 56 year-old man was admitted with severe headache and high grade fever. Computed tomography demonstrated intracranial hemorrhage. Cerebral mycotic aneurysm was detected at left distal middle cerebral artery by cerebral angiography. Infective endocarditis and mitral regurgitation were also diagnosed by echocardiography. He underwent cerebral mycotic aneurysmectomy after intensive antibiotics therapy, followed by successful mitral valve replacement. We review the literatures and discuss the problems of surgical management of infective endocarditis with cerebral mycotic aneurysm. PMID- 9223859 TI - [3 Cases of blunt chest trauma]. AB - 3 Cases of blunt chest trauma were reported, including aortic isthmus rupture, diaphragmatic injury and right main bronchial rupture. All cases were injured by traffic accidents. The first case was a 21-year-old man complicated with aortic injury and femoral bone fracture and died of intrapleural rupture before thoracotomy. The second case was a 49-year-old man who had right 11th rib fracture and he received emergent thoracotomy due to hemorrhagic shock. We found accidentally arterial massive bleeding by the diaphragmatic injury at thoracotomy and ligated directly. The third case was a 19-year-old man and he was unconscious on admission and rupture of the right main bronchus was found by bronchofiberscope and performed pneumectomy. One patient died and other two patients have survived. Blunt chest trauma sometimes lead to fatal damage of vital organs. So it is necessary to make early diagnosis and perform the emergent operations for life salvage. PMID- 9223860 TI - [Thoracoscopic cutting needle biopsy]. AB - We presented a new method of percutaneous cutting needle biopsy under the thoracoscope. Percutaneous cutting needles for lung tumor have been abandoned because the risk of hemorrhage, death, and pneumothorax is much greater with these instruments than the computed tomographic guided needle aspiration. On the other hand, thoracoscopic stapled lung wedge resection is easy to obtain a good sample for any lung disease, but should be needed two or three staples, and waste a time, in the case following lobectomy was needed for malignancy. This thoracoscopic cutting needle biopsy was easy to obtain a specimen for the frozened section diagnosis. Under the view of thoracoscope, percutaneous cutting needle biopsy is safety, saving the cost for staples and easy to control the bleeding from the lung. The indication of this thoracoscopic cutting needle biopsy is the case that could not obtain the precise diagnosis by the preoperative biopsy and imaging diagnosis might be suspicious a malignant tumor. PMID- 9223861 TI - [A case report of coronary artery bypass grafting (CABG) and graft replacement of the ascending aorta for coronary artery disease associated with dissection of the ascending aorta]. AB - A 64-year-old woman waiting for CABG for triple coronary artery disease admitted to our hospital due to chest oppression. One week after the admission, pericardiotomy was performed and blood in the pericardial effusion was found. Emergent chest CT scan revealed dissection of the ascending aorta (DeBakey II) with occluded false lumen. Because her condition was stable, concomitant operation of graft replacement of the ascending aorta and triple coronary artery bypass grafting were performed 3 months after the medical therapy. Her postoperative course was uneventful and the patient remains asymptomatic. PMID- 9223862 TI - [The leaflet fracture of a 19 mm SJM (HP) valve during valve insertion to the calcified small aortic annulus]. AB - Aortic valve replacement was undergone in a 70-year-old woman with calcified aortic stenosis. Calcified and fused tricuspid aortic valve was excised and a 21 mm sizer passed with some difficulty through the annulus. A 19 mm SJM HP serious was selected and the horizontal mattress suturing technique was used. The valve was held at the cuff near the hinge guard and seated into the annulus by the usual amount of pressure, however, one leaflet fracture was retrieved and the valve was removed. A 19 mm standard SJM valve was then successfully implanted. The potential factors of leaflet fracture suggested that the leaflet thickness of smaller sized SJM valves showed smaller values than other bileaflet valves in the same sized-valves. In addition, the SJM valve does not have an orifice stiffening ring like other bileaflet valves. So, needless to say direct pressure to the leaflet, indirect pressure might cause valve destruction during insertion. The abnormal pressure applied to the direction from one hinge guard to the other can introduce deforming of the orifice ring resulting in leaflet fracture like this case. The pressure directed to the perpendicular direction may introduce leaflet escape due to the deforming of the valve orifice. PMID- 9223863 TI - [Aortic valve replacement with the Toronto stentless porcine valve in a patient with clipping for cerebral arterial aneurysm]. AB - A 48-year-old man, who had a cerebral arterial aneurysm, was admitted in our institution for operation of aortic valve stenosis. At first, he underwent clipping for cerebral arterial aneurysm under precise management of his hemodynamic condition. After the clipping operation, we performed aortic valve replacement with the Toronto stentless porcine valve because no anticoagulant therapy was ideal for patient with cerebrovascular disease and larger effective orifice area was preferable for stenotic aortic annulus. By means of echocardiography, mean pressure gradient of the aoric valve decreased from 42 mmHg to 22 mmHg after the valve operation. He was discharged from the hospital on the 23rd postoperative day, and he has been doing well without thromboembolic events and bleeding complications for five postoperative months. This experience suggest that the Toronto stentless porcine valve might be one of the valve of choice for patients with aortic valve disease and cerebrovascular disease. PMID- 9223864 TI - [A case report of sudden death after Bentall's operation (Piehler's modification) due to left coronary occlusion]. AB - We encountered a rare complication after Bentall's operation (Piehler's modification) with the left coronary artery occlusion. A 56-year-old man admitted to our hospital to get the aortic reconstruction operation for the chronic dissecting aortic aneurysm (DeBakey type I) with 7 cm in diameter. We performed Piehler's modification procedure using a 26 mm and 8 mm woven Dacron tubes coated with gelatin, and 23 mm prosthetic valve (St. Jude Medical). He got well after 6 day intubation until he complained anterior chest pain suddenly on 22 postoperative day. He showed ventricular fibrillation on the second attack requiring resuscitation procedures. Emergent angiogram showed total occlusion of the left coronary artery. The percutaneous transluminal coronary angioplasty was performed only with temporal success. Autopsy revealed no thrombus in the grafts and no kinking of the tubes. We might have to use more anticoagulants during the respirator assisted period when the APTT was naturally prolonged from 120 to 160 seconds. PMID- 9223865 TI - [A case of atrial septal defect complicated by persistent left superior vena cava in whom hemophagocytic syndrome appeared after operation]. AB - The patient was a 22-year-old woman and suffered from atrial septal defect (ASD) complicated by persistent left superior vena cava (PLSVT). The patient underwent a patch closure using calf pericardium. Despite favorable progress immediately after the operation, fever, enlarged cervical lymph nodes, and leukocytopenia appeared beginning day 8 after the operation. Based on the results of cytodiagnosis of myeloaspiration specimen, the diagnosis of hemophagocytic syndrome (HPS) was made. Since an elevation of 2-5 oligo A synthetase (2-5 AS) levels was observed, the patient was diagnosed to suffer from virus-associated hemophagocytic syndrome (VAHS), and steroid was administered. The patient exhibited a good response to the treatment. The fever declined 3 days after the start of administration. Leukocyte counts were also increased gradually. The patient condition was improved and she was discharged from the hospital on day 28. At present, seven months after the operation, there is no evidence of recurrence and the patient's condition is still monitored. PMID- 9223866 TI - [Mitral valve replacement and closure of ventricular septal defect in the first two months of life]. AB - A 2-month-old female infant weighing 4.2 kg was admitted with severe congestive heart failure and respiratory distress. The patient was operated upon under a diagnosis of severe congenital mitral regurgitation and ventricular septal defect. There was thickening of the anterior leaflet and short thickened chordae. The valve was judged not amenable to repair and it was replaced with a 17 mm St. Jude Medical valve. She was extubated on the 5th postoperative day. Post operative anti-coagulant therapy was initiated with warfarin potassium, dipyridamole and ticlopidine hydrochloride. The postoperative course was uneventful and the patient is doing well 9 months after the operation. PMID- 9223867 TI - [A case of chronic expanding hematoma resected 37 years after lobectomy]. AB - A 65-year-old man was admitted for evaluation of slowly growing mass shadow in the right lower lung field on chest X-ray film. He had undergone right upper lobectomy for pulmonary tuberculosis 37 years before. As we could not rule out a suspicion of mediastinal tumor, thracotomy was performed. Histological examination of the resected mass showed a hematoma surrounded by dense fibrotic tissue. And the mass was resultantly diagnosed as chronic expanding hematoma. The mechanism of chronic expanding hematoma is unclear. In our review of the Japanese literatures, only 10 cases including this one, resected more than twenty years after the initial trauma or operation, could be found. PMID- 9223868 TI - [A child of malignant lymphoma diagnosed by transbronchial aspiration cytology under general anesthesia with a laryngeal mask]. AB - A 8-year-old male with Wiskott-Aldrich syndrome was admitted to our hospital because of left hilar swelling on a chest roentgeonogram. Bronchofiberscopy and transbronchial aspiration cytology (TBAC) was performed under general anesthesia with a laryngeal mask. The specimen obtained by TBAC was immediately stained and diagnosed as malignant lymphoma. One month later, thoracotomy was performed in order to get more detail diagnosis, which is necessary to determine the regimen of chemotherapy, and the nodal specimen were diagnosed as Hodgkin's disease. The ABVD therapy was performed which induced the complete remission. The laryngeal mask is a useful device for bronchofiberscopy in children. PMID- 9223869 TI - [A case of successful endoscopic treatment for traumatic bronchial rupture]. AB - Successful endoscopic treatment with fibrin glue for traumatic bronchial rupture was reported. 21-year-old male, who was injured in a traffic accident was brought to ICU in our hospital. Six days after injury, bronchoscopic findings showed a rupture from right truncus intermedius to lower bronchus (B8). We preferred an endoscopic treatment with fibrin glue because of poor respiratory function caused by lung contusion as a complication and those resulted in successfully healing. In general, basic strategy for traumatic bronchial rupture is the earlier diagnosis and surgical treatment. But in a case of high risk for operation, we recommend endoscopic treatment with fibrin glue might be useful. PMID- 9223870 TI - [A case of advanced lung cancer with long time survival, involving direct invasion to the liver]. AB - A 64-year-old male was admitted to our hospital complaining of right back pain. He had squamous cell carcinoma located at lower lobe of right lung involving the diaphragm and liver, pointed out by chest CT. Main tumor, the lateral chest wall, the posterior side of the diaphragm and the posterior side of the liver (S7) were resected. This patient has survived without any recurrences for 5 years and 10 months after operation. It is important not to hesitate in performing combined resections, even if the advanced lung cancer has invaded into the liver. Because the long-term survival might be achieved. PMID- 9223871 TI - [A case of schwannoma arising in the brachial plexus with intrathoracic extension]. AB - A case of benign schwannoma originating from the lowest trunk of the left brachial plexus with intrathoracic extension was reported. Intrathoracic growth of a schwannoma of the brachial plexus has been reported in only 3 cases in the literature. The patient was a 53-year-old man and he was first found to have an abnormal shadow at the left lung apex on the chest roentogenogram. We performed a thoracotomy through the left third intercostal space with an axillary skin incision and removed the tumor completely. There was no neurological problem postoperatively PMID- 9223872 TI - [A carcinoma arising from benign pleomorphic adenoma of the trachea]. AB - A 69-year-old female complained of husky voice and dyspnea. Bronchofiberscopic examination revealed a tumor at the left side of the trachea, which obstructed approximately 80% of the tracheal lumen. Benign pleomorphic adenoma was diagnosed by biopsy. The tumor was removed by circumferenctial resection of the trachea with partial sternotomy adding to the 3 th intercostal transverse resection. Historical finding of the resected specimen revealed a carcinoma arising from benign pleomorphic adenoma of the trachea and residue of malignancy at the margin of the trachea and esophagus. Additional radiotherapy was performed (60 Gy). The postoperative course was uneventful for 4 months and one year. PMID- 9223874 TI - [The effect of low-dose prostaglandin E1 on serum and urinary fluoride concentrations in patients anesthetized with sevoflurane]. AB - We investigated the effects of low-dose prostaglandin E1 (PGE1) on serum and urinary concentrations of inorganic fluoride in 39 adult patients undergoing upper abdominal surgery. Anesthesia was maintained with a combination of N2O-O2 sevoflurane and thoracic epidural anesthesia. Twenty-two patients received infusion of PGE1 at a rate of 0.02 micrograms.kg-1.min-1 throughout surgery. Seventeen patients served as control by not receiving PGE1. Serum inorganic fluoride concentrations (FB) were determined before the induction of anesthesia and 0, 2 and 24 hours after the end of anesthesia. Urinary inorganic fluoride concentrations (FU) were determined before the induction of anesthesia, and 0, 24 and 48 hours after the end of anesthesia. These was no difference between PGE1 group and control group in anesthetic dose (MAC hours) of sevoflurane. In both groups, FB peaked at the end of anesthesia. In PGE1 group, UB peaked at the end of anesthesia, while in control group, it peaked 24 hours after anesthesia. There were differences between groups neither in FB nor in FU throughout the study period. The relationships between anesthetic dose and fluoride concentrations, however, differed significantly between the groups. In control group FB values of 0, 2 and 24 hours after anesthesia correlated positively with MAC hours, respectively, while in PGE1 group they did not. Similarly in control group, FU values of 24 and 48 hours after anesthesia correlated positively with MAC hours, respectively, while in PGE1 group, they did not. Thus in patients receiving high dose sevoflurane, FB and FU tended to be lower in PGE1 group than in control group. In contrast, in PGE1 group, urinary excretion of fluoride during surgery correlated positively with MAC hours, while in control group, it did not. Urinary fluoride excretion during surgery was significantly greater in PGE1 group than in control group. These results suggested that PGE1 might prevent elevation of serum and urinary fluoride concentrations in patients receiving high-dose sevoflurane. This effect might result from enhanced urinary excretion of fluoride with PGE1. PMID- 9223875 TI - [Comparison of epidural fentanyl versus bupivacaine in maintaining intraoperative liver function]. AB - This study was undertaken to evaluate epidural anesthesia in maintaining of intraoperative liver function. Forty six ASA I and II patients undergoing elective open abdominal surgery were randomly allocated to 4 groups (I-IV) ; I (n = 12) : Epidural fentanyl + A, II (n = 12): Epidural fentanyl + B, III (n = 10) : Epidural bupivacaine (0.25%) + A, IV (n = 12) : Epidural bupivacaine + B. A and B patients were hydrated with acetated Ringer at a speed of 20, 10 ml.kg-1.h-1 for the first hour, 10, 5 ml.kg-1.h-1 for the next hour, then 5, 5 ml.kg-1. h-1 respectively, ICG 15 min values (ICG K: indocyanine green disappearance rate, R: retention rate) and AKBR (arterial ketone body ratio) were measured before induction, 10 min after intubation, 20 and 60 min after making a incision and 60 min after extubation. ICG K was larger and ICG R was smaller in I and II than in III and IV. AKBR was higher in I and III than in IV. AKBR in II showed a middle value between I, II and IV. Intraoperative liver function was better-maintained with epidural fentanyl regardless of difference of hydration. The results demonstrated that epidural fentanyl is one of the effective methods of anesthesia in maintaining intraoperative liver function. PMID- 9223876 TI - [Placental transfer of vecuronium administered with priming principle regimen in patients undergoing cesarean section]. AB - Eight women having cesarean section under general anesthesia received vecuronium (VCB) 0.01 mg.kg-1 as a priming dose, followed 180 s later by 0.11 mg.kg-1 as an intubation dose. Subsequently, VCB concentrations of umbilical venous and maternal arterial blood at delivery were assayed. The time from the injection of intubation dose to delivery was 283 +/- 55 (mean +/- SD) s. Umbilical and maternal VCB concentrations at delivery were 79.4 +/-36.1 1ng.ml-1 (UV) and 1258.3 +/- 464.1 ng.ml-1 (MA). respectively. Thus, the umbilical venous to maternal arterial VCB concentration ratio (UV.MA-1) was 0.07 +/- 0.02. One-min and 5-min Apgar scores were 8-9 and 9-10, respectively. Judging from previous reports concerning VCB administration during cesarean section, total of VCB 0.12 mg.kg-1 may be an overdose. We concluded therefore that of total VCB 0.10 mg.kg-1 seems to be an appropriate dose for cesarean section. PMID- 9223877 TI - [Effects of midazolam and propofol on dopamine release from rat striatal slice using a fast-cyclic voltammetry system]. AB - In order to evaluate the difference in pharmacological mechanism between midazolam and propofol, we focused on the interaction between dopaminergic and GABAergic neurons in the striatum because of its important role in the regulation of motor system and arousal response, and examined these inhibitory effects on dopamine (DA) release induced by high-K from the rat striatal slice using the fast-cyclic voltammetry method. Between 0.1 and 200 nmol, the standard curve of DA was linear. The peak and the sensitivity of DA oxidation were different from those of norepinephrine and DA main metabolites. The dosages between 0.1 and 10 micro M of propofol significantly blocked the high-K evoked DA release, although the dosage of larger than 50 micro M of propofol potentiated DA release. In case of midazolam, the dosages between 0.1 and 50 micro M markedly suppressed DA release induced by high-K in a dose-dependent manner. The recovery time of DA release after removal of midazolam from incubation medium was longer than that seen in the treatment of propofol. In conclusion, propofol and midazolam inhibited high-K evoked DA release from striatal slice, although these efficacies were dissimilar. Furthermore the pharmacological effects of larger dosage of propofol was different from those obtained by its smaller dosages. These results suggest that the anesthetic actions of propofol and midazolam are partially related to inhibition of DA neuron A1 activity and that the excitatory symptom induced by a larger dose of propofol may be related to its potantiation of DA release. PMID- 9223878 TI - [Relationship between the dose of lidocaine administered endotracheally in small children and the serum lidocaine level--recommended dose prior to endotracheal intubation]. AB - To determine the optimum dose of lidocaine administered in the trachea prior to endotracheal intubation, we divided 102 surgical patients 3-5 years of age into 3 groups, i.e., group 1; 1 mg.kg-1 was sprayed in the trachea, group 2; 2 mg.kg-1 was sprayed in the trachea, group 3; 1 mg.kg-1 was sprayed in the trachea, and 1 mg.kg-1 in the pharynx and the oral cavity simultaneously. The venous serum concentration of lidocaine was measured two times either 1.5, 3, 5, 10, 15, 20, 30 or 45 min after the spray in each case. In group 1, the mean concentration of lidocaine reached the maximum of 1.05 micrograms.ml-1 at 5 min and decreased gradually after that with a small inter-individual variation. In group 2, after reaching the mean maximum concentration of 3.51 micrograms.ml-1 at 3 min, the serum level dropped quickly and then gradually decreased. There were a few cases where serum level was over 7 micrograms.ml-1. In group 2, after reaching the mean maximum concentration of 1.38 micrograms.ml-1 at 5 min, the serum level decreased more slowly, suggesting a slow absorption from the pharynx and/or the oral cavity. We conclude that the recommended dose of lidocaine for endotracheal administration is less than 2 mg.kg-1. PMID- 9223879 TI - [Adrenergic receptor and alpha 2 agonist--2: Structure-function relationship of adrenoceptors]. AB - Recombinant DNA experiments using chimeric receptors containing portions of alpha 2 and beta 2 adrenoceptors demonstrated structure-function relationships of adrenoceptors. The seventh transmembrane domain determines the subtype ligand binding specificity between alpha 2 and beta 2 adrenoceptors. A further investigation by mutagenesis suggests that a direct interaction between subtype specific ligands and specific amino acids such as Phe (412) and Asn (312) in the seventh transmembrane domain of the alpha 2 and beta 2 adrenoceptors respectively. The third cytoplasmic loop is responsible for determining the specificity of interactions between the receptor and G protein. Recombinant DNA technology also demonstrated that seven transmembrane domains of adrenoceptors have a counterclockwise arrangement when viewed from the outside of the cell. PMID- 9223880 TI - [Effect of injection speed on sensory blockade in spinal anesthesia with 0.5% hyperbaric tetracaine]. AB - We investigation the effect of injection speed on sensory blockade in spinal anesthesia. Forty two female patients, scheduled for total abdominal hysterectomy, were allocated randomly to 3 groups of 14 each according to the injection speed of 0.5% hyperbaric tetracaine: Group F (fast; injection speed > or = 0.2 ml.s-1), Group M (moderate; 0.1 < injection speed < 0.2 ml.s-1) and Group S (slow; injection speed < or = 0.1 ml.s-1). Spinal puncture was performed via the median approach at the L3-4 interspace with the patient in a lateral position. The maximum level of sensory blockade was assessed by means of the pin prick method in the midline 3, 5, 10, 20, 30, 60 minutes after injection. In Group F, the level of sensory blockade became higher quickly (within 5 min), but anesthetic effects were not so satisfactory. And, in this group, there were more patients with dyspnea than in other groups. We speculate that the turbulence made by fast injection in subarachnoid space caused unsatisfactory effects. In Group S, anesthetic level was becoming higher also 20 or 30 min after injection. The fixation of anesthetics requires about 30 min. In our opinion, anesthetics injected slowly were diluted less by CSF, and the actual baricity of them was higher, and this made the difference within 30 min. In Group M, anesthetic effects and patient's condition were stable. We suppose that this injection speed (0.1-0.2 ml.s-1) is suitable for spinal anesthesia. PMID- 9223881 TI - [Living related liver transplantation for patients with ornithine transcarbamylase deficiency]. AB - Ornithine transcarbamylase deficiency (OTCD) is an inborn error of urea synthesis inherited as an X-linked trait, a clinical manifestation of which is a repeated episodes of hyperammonemic coma. Recently, liver transplantations have been performed in these patients in the USA and Europe. We experienced the anesthetic managements of liver transplantations in two OTCD patients, who had been suffering from several episodes of hyperammonemic decompensation despite a restricted protein diet with administration of sodium benzoate. Anesthesia was induced and maintained with a combination of fentanyl and midazolam in both cases. Their postoperative courses were good without any neurological damages, though one patient had hyperammonemic attack during the operation. PMID- 9223882 TI - [Preanesthetic meals in elective surgical patients]. AB - This study investigated the effect of preanesthetic meals on the volume and pH of gastric contents in forty elective surgical patients ranging in ages from 20 to 60 years. Twenty patients who were given either isotonic beverage 250 ml or apple juice 250 ml on the morning of the operative day were subjected as control group and twenty patients of the breakfast group took two slices of bread with the above drink. About seven hours following drinking and feeding, the mean values of gastric volume were 20.9 +/- 18.3 ml in the control group and 19.2 +/- 16.3 ml in the breakfast group. The mean values of gastric pH were 4.3 +/- 2.3 in the drink group and 4.6 +/- 2.3 in the breakfast group. There were no significant differences in the gastric volume and pH between the two groups. However, very small amount of the bread was detected in the gastric fluid of three patients in the breakfast group. As preanesthetic drinking and feeding are advantageous for reducing the anxieties of preoperative patients and also for their nutrition during operation, it is encouraging that eating two slices of bread did not induce a significant effect of gastric volume or pH. The minute fragment of bread seems to have no clinically significant effect. PMID- 9223883 TI - [Efficacy of lidocaine tape for venous cannulation in children]. AB - We evaluated whether lidocaine tape (3 X 5 cm, contained 18 mg of lidocaine) could reduce pain caused by venous cannulation in children during anesthetic induction. One hundred and thirty-five children scheduled for elective surgery were randomly assigned to three groups according to the application time of the tape (30 min, 60 min and 120 min). Pain assessment was made by using our pain score (0: no response, 1: slight agitation, 2: strong agitation, at the venous cannulation). The effect of pain reduction (pain score 0 and 1) was found in 81 % of all patients. Especially in the group of 120 min, the effect was remarkable. Only 6.7% of all patients had slight adverse effects including skin redness and itching. In another 25 patients (weighing 3.5-30 kg), plasma concentration of lidocaine were measured 120 min after application of a piece of this tape. Arterial blood was sampled at 30 and 120 min after the tape was removed. Plasma lidocaine levels were always below 0.8 mcg.ml-1 In conclusion, the lidocaine tape may be useful and safely applicable for venous cannulation in children. PMID- 9223884 TI - [Epidural buprenorphine and epidural droperidol for post-operative nausea or vomiting]. AB - For 120 young patients who had undergone reconstruction of anterior cruciate ligament of the knee, we investigated the effect of epidural administration of buprenorphine on the incidence of nausea or vomiting, and the anti-emetic effect of epidural administration of droperidol. In the group who had received bolus injection of buprenorphine 0.1 mg, nausea or vomiting occurred early most often, and the injection was not useful to prolong analgesic effect. In the another group who had received continuous epidural infusion of buprenorphine 0.3 mg, nausea or vomiting occurred late. Bolus injection of droperidol 2.5 mg was not useful to prevent nausea or vomiting caused by continuous epidural infusion of buprenorphine. While, continuous infusion of droperidol 5 mg was effective in decreasing nausea or vomiting caused by continuous epidural infusion of buprenorphine. In conclusion, continuous epidural administration of droperidol is useful to prevent nausea or vomiting. PMID- 9223885 TI - [The use of low dose midazolam for the management of spinal anesthesia]. AB - The purpose of this study was to evaluate the effects of low dose midazolam (MZ) on memories and ease of management of spinal anesthesia. The low doses of MZ were administered to 70 patients (ASA 1-2), of whom 37 patients were premedicated with atropine sulfate 0.5 mg and pethidine hydrochloride (group P), with 33 patients receiving no premedication (group N). Double blind randomized trials were conducted with the doses of MZ (0, 0.03, 0.06 mg.kg-1), and MZ was administered i.v. to patients just prior to spinal puncture. The short-term and long-term memories were impaired after administration of MZ in both groups. We conclude that MZ appears to be a suitable replacement for other benzodiazepines for relieving anxiety of patients during surgery. PMID- 9223886 TI - [Effect of hemodilution with dextran and albumin on activated clotting time (ACT)]. AB - Hemodilution with colloid solution is known to influence blood coagulation. To evaluate the effectiveness of hemodilution on ACT, 156 patients undergoing elective coronary artery bypass grafting were studied. Patients were divided into three groups (Group I of 52: without hemodilution, Group II of 52: normovolemic hemodilution with Dextran 40, Group III of 52: albumin and lactated Ringer solution). Systemic heparin was given (300 IU.kg-1. ACT was measured by Hemochron 400 and maintained higher than 400 seconds with supplemental dose of heparin. Sampling was performed after induction in Group I, after hemodilution in other groups, after heparin administration, was after start of CPB. More supplemental dose of heparin were required to achieve the lower limit of ACT (400 seconds) in the group I. The ACT at any sampling point was significantly higher in the group II and III than in the group I. In conclusion, hemodilution with dextran and albumin increased ACT values and heparin effect. PMID- 9223887 TI - [Anesthetic management for a patient with pure autonomic failure]. AB - Pure autonomic failure is characterized by orthostatic hypotension, sweating disorder, urinary incontinence, and syncope. A 64 year-old man with pure autonomia failure was scheduled for suprapubic prostatectomy. We monitoring direct arterial pressure and inserted pulmonary artery catheter prior to the induction of anesthesia. General anesthesia was induced with diazepam 10 mg, fentanyl 0.3 mg, and vecuronium 8 mg for tracheal intubation. Anesthesia was maintained with sevoflurane (0.2-1.5%), 60% nitrous oxide in oxygen supplemented with intermittent epidural anesthesia. During anesthesia, blood loss was immediately replaced with banked blood because autonomic failure could not compensate hypovolemia well. Epidural anesthesia in this patient was considered to cause less hypotension than in patients with normal autonomic function. Therefore, we think epidural anesthesia is a useful anesthesia method for patients with pure autonomic failure. The emergence from anesthesia was smooth and no complications were seen during the perioperative period. PMID- 9223888 TI - [Evaluation of postoperative hypoxemia after spinal anesthesia with a pulse oximeter]. AB - We investigated postoperative hypoxemia after spinal anesthesia in 24 adult patients by monitoring SPO2 with a pulse oximeter for 5 hours in the ward. The patients received spinal anesthesia with 0.5% hyperbaric tetracaine and sedated with midazolam during operation. Twelve patients inhaled 3 l.min-1 oxygen via a mask for initial 3 hours, and the other breathed room air all the time. In two of the patients breathing room air, SPO2 decreased to less than 94%. In patients inhaling oxygen, SPO2 was maintained above 98% but decreased significantly after stopping oxygen inhalation (below 94% in five patients). The postoperative SPO2 value correlated significantly with BMI (body mass index) and preanesthetic SPO2, but not with the dose of midazolam or supplemental pentazocine, and postoperative conscious states. In conclusion, there are some incidences of postoperative hypoxemia after spinal anesthesia irrespectively of the dose of midazolam used intraoperatively. Inhalation of 3 l.min-1 oxygen via a mask is sufficient to prevent such postoperative hypoxemia. PMID- 9223889 TI - [Acute angle-closure glaucoma after total hip replacement surgery]. AB - Acute angle-closure glaucoma is a rare complication of surgery. We experienced a case of postoperative acute glaucoma after total hip replacement under general anesthesia. A 49-year-old female without signs or symptoms of glaucoma was premedicated with the intramuscular administration of secobarbital, atropine and ranitidine. Following rapid induction with thiopental and vecuronium, anesthesia was maintained with N2O-O2-sevoflurane. PGE1 was administered intravenously for induced hypotension during the surgery. Hemorrhagic shock with a systolic blood pressure of 60 mmHg continued for 15 min during the surgery. Large amounts of fluid and ephedrine were required for treating this hypotensive episode. Vecuronium was reversed by bolus injection of neostigmine and atropine at the end of surgery. Soon after recovery from anesthesia, she complained of pain and blurred vision in her both eyes. The consulting ophthalmologist made a diagnosis of acute glaucoma due to high intraocular pressure (IOP). Treatment with glycerol and pilocarpine had no effect on the elevated IOP. The laser iridotomy performed on her at 5th and 7th post-operative days improved her vision completely. The post-operative glaucoma may cause serious permanent loss of vision. An early diagnosis of this post-operative complication and its treatment with drugs and surgery should be emphasized. PMID- 9223890 TI - [Severity and prognosis of congenital diaphragmatic hernia from the viewpoint of perioperative respiratory function]. AB - We studied severity and prognosis of congenital diaphragmatic hernia (CDH) by using preductal arterial blood gas analysis (BGA) and pulmonary function tests (PFTs) in 29 newborn infants. CDH was diagnosed within 24 hours of life, and surgical repair was performed through an abdominal approach after a period of stabilization. The infants were classified into the following three groups based on the highest preoperative alveolar-arterial oxygen tension difference (A - aDO2) and the lowest arterial carbon dioxide pressure (PaCO2) values; Group A (n = 15) : A - aDO2 < 500 mmHg, PaCO2 < 40 mmHg, Group B (n = 7) : A - aDO2 > or = 500 mmHg, PaCO2 < 40 mmHg, Group C (n = 7) : A - aDO2 > or = 500 mmHg, PaCO2 > or = 40 mmHg. Furthermore, the patients were classified into the following three groups based on the preoperative respiratory system compliance (Crs) and forced vital capacity (FVC) values; Group D (n = 8) : Crs < 0.5 ml.cmH2O-1.kg-1, FVC < 10 ml.kg-1, Group E (n = 4) : Crs < 0.5 ml.cmH2O-1.kg-1, FVC > or = 10 ml.kg-1, Group F (n = 17) : Crs > or = 0.5 ml.cmH2O-1.kg-1, FVC > or = 10 ml.kg-1. The mortality in the Group C was significantly higher than in the Group A and B, and the preoperative Crs and FVC values in the Group C were significantly lower than the other groups. The mortality in the Group D and E were significantly higher than the Group F. In conclusion, it is suggested that the preoperative Crs value less than 0.5 ml.cmH2O-1.kg-1 indicates severe pulmonary hypoplasia and is critical for survival. PMID- 9223891 TI - [The effect of prophylactic intravenous diltiazem drip infusion on myocardial ischemia during noncardiac surgery]. AB - We evaluated the effect of intravenous diltiazem infusion in 105 noncardiac surgical patients. Subjects were elective surgical patients with coronary artery disease and coronary risk factors which were hypertension (WHO standards), diabetes mellitus, hyperlipemia (total cholesterol > or = 220 mg.dl-1), obesity (body mass index : male > or = 26 kg.m-2, female > or = 25) and old age (70 years old or above). The prophylactic intravenous diltiazem infusion (1.0 micrograms.kg 1.min-1) was started immediately after induction of general anesthesia or epidural analgesia and continued until the end of operation. All patients were monitored by ST trend graph during anesthesia, and ischemia pattern was defined as > or = 1 mm ST changes and lasting over 1 min. Ischemic ST-T changes were noted in 4 cases in the operating room. ST depression was noted in 2 cases before starting anesthesia and these 2 cases showed improvement with diltiazem infusion lasting until the end of operation. ST-T changes were noted in 2 cases during surgery and these 2 cases showed improvement with diltiazem isosorbide dinitrate. We conclude that prophylactic intravenous diltiazem infusion may prevent ischemia during noncardiac surgery. PMID- 9223892 TI - [Minimally invasive technique for mitral valve repair--the importance of intraoperative transesophageal echocardiography]. AB - A 65-year-old male with mitral regurgitation was scheduled for the mitral valve repair by means of minimally invasive technique. The technique, which requires a 10-cm right parasternal incision, offers minimal discomfort, less postoperative pain, quick functional recovery and excellent cosmetic healing. However, the intraoperative monitoring by the transesophageal echocardiography was required to evaluate the mitral valve function and to confirm the complete removal of air from the apex of the left ventricle and right upper pulmonary vein due to the small operative field. In addition, the monitoring of the short-axis view of the left ventricle makes it possible to evaluate and to confirm the preload and the left ventricular contractility. We considered that the transesophageal echocardiography is essential and useful for the minimally invasive mitral valve repair. PMID- 9223893 TI - [Transmitral flow analysis during the coronary artery bypass surgery]. AB - Left ventricular diastolic function (LVDF) derived from the analysis of Doppler transmitral flow (TMF) has been analyzed. Since little is known about changes of LVDF during cardiac surgery, our aim of this study is to investigate TMF as a predictor of LVDF during the coronary artery bypass surgery (CAB). Twenty-eight patients were enrolled in this study and divided into two groups depending on the ratio of peak early filling (E) and atrial (A) velocity (E/A > or = 1; good LVDF, E/A < 1; impaired LVDF) measured by transesophageal echocardiography after induction of anesthesia. In both groups, E/A and E decreased significantly during internal mammalian artery dissection compared with after induction, suggesting left ventricular dysfunction. Especially in impaired LVDF group, peak A velocity did not increase in spite of the significant reduction in cardiac output and atrial contributions failed to compensate hemodynamics in this setting. More detailed features of cardiac function might be obtained by TMF analysis during CAB. PMID- 9223894 TI - [The efficiency of a new porous type leukocyte removal filter for red cell blood components, Terumo Imuguard III-RC, in the rapid transfusion conditions]. AB - We evaluated the efficiency of a new porous type leukocyte removal filter for red cell blood components, Terumo Imuguard III-RC, in the rapid transfusion conditions. One leukocyte removal filter was used for 2 units of RC-M.A.P (red cell mannitol-adenine-phosphate). Filtration methods employed were gravity infusion, high pressure infusion (300 mmHg), pumping infusion and 20 ml.min-1 infusion under high pressure (300 mmHg). Blood samples were taken before and after the filtration to measure white blood cell (WBC), red blood cell (RBC) and platelet content. Blood samples before filtration and after filtration with WBC excluded, were examined by automated hematology analyzer (Coulter counter STKS Retic). WBC after filtration was counted by the hemacytometer method using Nageotte Chamber. The removal rate of WBC was found to be more than 99.99% and residual WBC content was less than 4 x 10(4) with every method. The recovery rate of RBC was not significantly decreased in all filtration methods. The removal rate of platelet was equal in all filtration methods. In conclusion, Imuguard III RC could be useful for effective homologous blood transfusion. PMID- 9223895 TI - [A psychological burden on patients with clinical trials during anesthesia]. AB - We investigated with questionnaires, psychological outline of 30 patients, who were requested to participate in clinical trials of anti-tachycardia drugs during anesthesia. Although 14 patients consented to the trial, the consent was not based on adequate understanding or volunteerism in 3 patients. Nine patients of the consented group were anxious about the possible use of the trial drug. Eight patients of the rejected group felt anxiety on surgery and anesthesia, which was the most common reason for rejection. Forty % of refused patients felt a guilty conscience or embarrassed. Although we tried to obtain patients' consent following governmental and institutional regulations and guidelines, not only the consented but also the refused patients suffer from psychological burden with the clinical trial. It is of concern that recruitment to the trial enhances anxiety of the patients as they already feel uneasiness, unrest, and insecurity facing anesthesia and surgery. To avoid entry of less informed or unwilling patients to the clinical trial, we must secure patients' veto, and recruitment should be performed by clinicians who are not involved in anesthesia practice. PMID- 9223896 TI - Malakoplakia of the appendix. AB - A case of a malakoplakia of the appendix in a sixty-year-old woman is reported. No involvement of the organs or important general diseases were present and the patient, after the appendicectomy, promptly recovered. Only three cases of malakoplakia of the appendix have previously been described in the literature and they are briefly summarized. PMID- 9223897 TI - Tobacco wars. PMID- 9223898 TI - Allergen immunotherapy. PMID- 9223899 TI - Informing the cancer patient. PMID- 9223900 TI - U.S. Supreme Court and Federal Trade Commission determine medicine is not a profession. Decision affects patient care. PMID- 9223901 TI - Radiology quiz #13. Lymphoma. PMID- 9223902 TI - Medical malpractice. The defense perspective. PMID- 9223903 TI - Medical malpractice. The plaintiff's perspective. PMID- 9223905 TI - Disease management as part of a population-based approach to physician directed medical care. PMID- 9223904 TI - Tobacco use among Missouri high school students, 1995. PMID- 9223906 TI - Outcomes measurement as part of a population-based approach to physician directed medical care. PMID- 9223907 TI - Gamma Knife radiosurgery: indications, techniques, and results in 200 patients treated at the Midwest Gamma Knife Center. AB - During radiosurgery, a single high dose of ionizing radiation is delivered with exquisite precision to a radiographically defined intracranial target. More than 60,000 patients have been treated worldwide since the first Gamma Knife was installed in Stockholm in 1968. Indications include arteriovenous malformation, cerebral metastasis, acoustic neuroma, meningioma, malignant glioma, and trigeminal neuralgia. Two hundred patients have been treated at the Midwest Gamma Knife Center in the first 21 months of operation beginning in September 1994. PMID- 9223908 TI - Why be just a doctor when you can be a spin doctor? PMID- 9223909 TI - Aetiological and histological classification of sialadenitis. PMID- 9223910 TI - Contribution of the Italian scientists to the ISDQP. International Society of Diagnostic Quantitative Pathology. PMID- 9223912 TI - Hypoglycemia in small-for-gestational-age neonates'. AB - In this prospective study, we investigated the frequency of hypoglycemia and the proper intervals for screening in small-for-gestational-age (SGA) neonates. We determined test-strip blood glucose values at two, three, six, 12, 24 and 48 hours of age in 25 SGA and 16 appropriate-for-gestational-age (AGA) infants who were born after 37 completed gestational weeks at the Obstetrics Clinics of Sisli Etfal Hospital. Serum glucose determination was immediately done if a test-strip value was less than 40 mg/dl. The frequency of hypoglycemia in SGA neonates was significantly higher (p:0097) than in AGA neonates. The first three hours, the sixth hour and the 48th hour postnatally were the most common hours for encountering hypoglycemia. Clinical signs were not true indicators of hypoglycemia. These data suggest that screening for hypoglycemia in SGA neonates should continue for 48 hours. PMID- 9223911 TI - Childhood tuberculosis: a report of 2,205 cases. AB - A total of 2,205 children with the diagnosis of tuberculosis followed at Hacettepe University Ihsan Dogramaci Children's Hospital between the years 1972 and 1992 were investigated in this retrospective study. Sixty-two percent of the patients were less than six years old. There was no gender preponderance among the cases. Thirty-four percent of the patients had predisposing diseases for tuberculosis, such as malnutrition, measles and pertussis. Thirty-five percent of the patients had a history of contact with a tuberculous patient. The most common type of tuberculosis was pulmonary (39%), but extrapulmonary cases outnumbered pulmonary tuberculosis patients. Thirty-three percent of the patients had been vaccinated with BCG. The Mantoux test was found positive in 62 percent of patients. The tuberculous culture was found to be positive in the sputum of 28 percent of pulmonary tuberculosis cases. Patients were treated with daily (59%) or intermittent (41%) antituberculous therapy regimens. The response to treatment was excellent. Relapse was observed in five cases. Mortality from tuberculosis was eight percent, 23 percent of which were from tuberculous meningitis. Although there was a decrease in the number of children with tuberculosis seen at our hospital between 1972 and 1992, the prevention and control of tuberculosis are still significant health problems for our country. PMID- 9223914 TI - Interferon-alpha therapy for refractory idiopathic thrombocytopenic purpura in children. AB - Interferon (IFN)-alpha therapy was presented as a possible treatment for refractory immune thrombocytopenic purpura (ITP). We used 3 x 10(6) U recombinant IFN alpha, subcutaneously three times a week, every other day, for a total of 12 doses, in five children with refractory ITP. Three patients showed no response and were classified with Type III. One patient gave a partial response, and the other one had been in remission for 71 weeks as of this writing. This is the longest remission period reported for IFN therapy. The patients were classified Types IIb and I, respectively. The therapy was well tolerated. We conclude that further studies are to needed evaluate IFN-alpha therapy in childhood ITP. PMID- 9223913 TI - Standardization of methacholine inhalation challenge. AB - Methacholine inhalation challenge has been shown to be an extremely useful diagnostic test. The purpose of this study was to document the reproducibility of methacholine inhalation challenge used in our clinic. In addition, we also examined the output of the delivery system, the stability of four-month-old methacholine solution, and the cumulative effect of methacholine. To document reproducibility, two identical challenges were performed in each of 19 asthmatic children. The influence of the previous doses of methacholine on bronchial response was examined by performing a third challenge with a single dose of methacholine in ten of these children. The remaining nine children were also tested for the third time with four-month-old methacholine solution to examine the stability of that solution. The output of the delivery system was assessed by measuring the change in weight of the nebulizer. Responses to methacholine were highly reproducible within one doubling dose interval. There was a small but significant cumulative effect of methacholine. Comparable results were obtained with newly prepared methacholine solution and four-month-old solution. The variability of the output of the same nebuliser was less than that of different nebulisers of the same model. The major clinical implication of our results is that our methacholine inhalation challenge procedure is standardized. This encourages more widespread use of this important diagnostic test for demonstration of airway hyper-responsiveness. PMID- 9223915 TI - Tumor necrosis factor-alpha and interleukin-1 beta levels in children with bacterial, tuberculous and aseptic meningitis. AB - Cerebrospinal fluid levels of tumor necrosis factor-alpha and interleukin-1 beta in 78 children with nonbacterial, bacterial and tuberculous meningitis, and in 34 control subjects were analyzed in order to evaluate the involvement of these cytokines in the pathogenesis of acute bacterial meningitis and their discriminative value between different etiologies of meningitis. Tumor necrosis factor-alpha and interleukin-1 beta levels were significantly higher in bacterial and tuberculous meningitis than in aseptic meningitis and in control subjects (p < 0.0001). There was no difference in the levels of tumor necrosis factor-alpha and interleukin-1 beta between nonbacterial meningitis and control groups. The finding that both tumor necrosis factor-alpha and interleukin-1 beta are increased in the cerebrospinal fluid of patients with bacterial and tuberculous meningitis whereas normal levels of these two cytokines have been found in patients with nonbacterial meningitis signifies that these cytokines may be used to differentiate between bacterial and nonbacterial meningitis. PMID- 9223916 TI - Neonatal septicemia in a neonatal intensive care unit. Results of four years. AB - The outcome of congenital and nosocomial septicemia has been documented in infants who were admitted to a neonatal intensive care unit over a four-year period. The overall incidence of neonatal septicemia in the neonatal intensive care unit was 5.4 percent. Common causes of neonatal septicemia were gram negative bacilli and staphylococci. Gram-positive microorganisms were the major causative agents for early-onset septicemia. Since the most common pathogen in cases of nosocomial sepsis was gram-negative bacillus, higher mortality rates were observed in nosocomial sepsis. The overall mortality rate in neonatal sepsis was 44.2 percent. The mortality rate in infants in whom nosocomial septicemia developed was significantly higher than in the infants in whom early-onset septicemia developed. However, as gram-negative colonization of the nursery recently changed to gram-positive microorganisms, the mortality rate is hoped to decrease. PMID- 9223917 TI - Comparison of growth, serum prealbumin, transferrin, IgG and amino acids of term infants fed breast mild or formula. AB - This study was designed to compare growth parameters, biochemical indices of protein metabolism and serum amino acid patterns in newborn infants fed either human milk (n: 22) or formula with protein and amino acid concentrations similar to human milk (n: 19) during the first two months of life. Growth parameters were normal and similar in all infants. Serum levels of transferrin and IgG were not significantly different, whereas prealbumin concentrations were found to be significantly higher in infants fed formula compared to those fed human milk. In the formula group, glutamic acid and cystine levels were significantly lower while phenylalanine and proline were significantly higher compared to the breastfed group. It was observed that feeding a formula with an amino acid pattern similar to that of human milk does not necessarily give rise to identical patterns of amino acids or indices of protein metabolism. PMID- 9223918 TI - Serum granulocyte-macrophage colony-stimulating factor and interleukin-1 levels in patients with repeated and non-repeated pulmonary infections. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) has a stimulating effect on erythroid, megakaryocytic and granulocyte-macrophage progenitors. Activated T lymphocytes, monocytes, fibroblasts and endothelial cells release GM CSF after stimulation by endotoxin and cytokines such as interleukin-1 (IL-1) and tumor necrosis factor. IL-1 is also released in response to bacterial infections and inflammation by phagocytic mononuclear cells. GM-CSF and IL-1 levels were examined in 10 patients with recurrent pulmonary infection (repeaters) and in 10 patients with acute pulmonary infection (non-repeaters) in the acute and recovery periods of infection. The mean serum GM-CSF and IL-1 levels of non-repeaters were significantly higher than those of repeaters in the acute period of infection (p < 0.002), but the same parameters of both groups were not different in the recovery period (p > 0.05). In addition, both the serum GM-CSF and IL-1 levels of repeaters and non-repeaters in the acute period of infection were higher than those in the recovery period (p < 0.05). There was a positive correlation between serum IL-1 and GM-CSF levels in non-repeaters (r = 0.746, p = 0.013), but no significant correlation between the same parameters in repeaters (r = 0.395, p = 0.259). In this study, we could not explain why the serum GM-CSF and IL-1 levels in repeaters did not increase as they did in non-repeaters; moreover, there was no significant correlation between serum IL-1 and GM-CSF levels in repeaters during the acute period of infection. These findings may be due to microenvironmental factors in bone marrow and/or other factors. PMID- 9223920 TI - Short- and intermediate-term efficacy of amiodarone in infants and children with cardiac arrhythmia. AB - The antiarrhythmic effect of amiodarone was examined in this retrospective study in a group of 20 patients with a mean age of 8.5 +/- 6.7 years (range 42 days to 20 years, median 9 years). Five patients with atrial flutter, one patient with atrial fibrillation, two patients with an intermediate rhythm between atrial flutter and atrial fibrillation, four patients with chaotic atrial tachycardia, three patients with atrioventricular reentry tachycardia, two patients with junctional ectopic tachycardia, and three patients with ventricular arrhythmias were treated with amiodarone. The mean duration of therapy was 9.1 +/- 12.3 months (range 1 month to 4 years). Before amiodarone treatment, 18 patients had been unresponsive to various antiarrhythmic drugs (range 1-8, median 2). Two patients received amiodarone as an initial therapy. It was administered orally at a dose of 10 mg/kg once per day for 10 days and then decreased to 5 mg/kg once per day. Amiodarone was effective in 16 patients (80%). Side effects occurred in three patients, including thyroid dysfunction, elevation of liver enzymes, and keratopathy. All side effects disappeared upon cessation of the therapy. We recommend amiodarone for the treatment of childhood arrhythmias, especially for the refractory types. PMID- 9223919 TI - Tubular markers in children with insulin-dependent diabetes mellitus. AB - The aim of the present study was to investigate the prevalence of tubular dysfunction and to assess the clinical significance of low-molecular-weight proteinuria and enzymuria in children with insulin-dependent diabetes mellitus (IDDM). N.acetyl-beta-D-glucosaminidase (NAG) and beta-microglobulin (beta 2 M) excretion was determined in 52 children with insulin-dependent diabetes mellitus and 28 controls. Patients were grouped according to the duration of diabetes: group 1 (n = 7): less than one year; group 2 (n = 27): one to five years; groups 3 (n = 18): greater than five years. Both parameters were significantly increased in groups 2 and 3 compared to controls. Urinary beta 2 M levels correlated significantly with albuminuria and HbA1C, while urinary NAG levels correlated only with HbA1C. Two to four samples were obtained from 35 of 52 diabetic patients in the study group at one-month intervals. Of these, 23 patients had elevated NAG levels, and 22 patients increased beta 2 M excretion. However, only six patients displayed persistent enzymuria, and nine low-molecular-weight proteinuria. The mean (SD) of coefficients of variation of each patient was 50.45 (+/-28.24) for NAG and 68.25 (+/-42.57) for beta 2 M excretion. We concluded that early tubular dysfunction and/or damage occurs in IDDM but is not established in the majority of children. PMID- 9223921 TI - Improvements in mother-child health indicators in Turkey. AB - Turkey has a young population as a result of high fertility and growth rates in the recent past. Thirty-five percent of the population is less than 15 years of age, and 25 percent of the population comprises reproductive-age woman. The latest estimate of the population growth rate was 19 per thousand for the 1990 1995 period. In recent decades dramatic declines in fertility rates have been noted. In the early 1970s, the overall fertility rate was approximately five children per woman, declining to 2.7 in 1993. The crude birth rate is currently estimated to be approximately 23 per thousand. The crude death rate has also declined from approximately 30 per thousand in the 1940s to 6.5 per thousand in the 1990s. Life expectancy at birth in the Turkey is 65.9 for males and 70.5 for females. The infant mortality rate in the 1960s was approximately 200 per thousand, declining to 67 per thousand during the 1985-1990 period, and 53 per thousand for the period 1988-1993. It was 48 per thousand in 1995 and 42.2 per thousand in 1996, according to the State Planning Organization. The infant mortality rate has declined by 35 percent in the last ten years. The mortality rate for children under five years of age was 113.5 per thousand between 1978 1983 and 60.9 per thousand in 1993; it is currently 50 per thousand. The maternal mortality rate was greater than 200/100,000 in 1995. During the last five years the proportion of women receiving antenatal care has increased from 43 to 63 percent. The proportion of safe deliveries was 76 percent. Thirty-nine percent of all deliveries occur at home. The important point here is that the proportion of unsafe deliveries assisted by traditional birth attendants is 24 percent. It is very obvious that during the last 15 years, maternal and child health (MCH), especially child health, has improved dramatically in Turkey. The improvement made in the last five years has been more marked. The improvement is closely related to the government's special interest, attention and efforts to prevent and identify the most common health problems and to overcome these problems with appropriate interventions. The government also pays special attention to the socio-economic priority areas of the country and initiates special health programs in these areas first, in order to reduce high regional differences in MCH indices. Despite these dramatic improvements, one great obstacle is the high maternal and infant mortality and morbidity rates in the country at the time of ratification of the European Social Charter. The success made so for in maternal and child health should not be ignored, but it must be realized that much still remains to be done to improve the MCH level further. PMID- 9223922 TI - Hepatitis-B-associated glomerulonephritis in children. AB - Three cases with hepatitis B virus (HBV)-related, biopsy-diagnosed glomerulopathies, one of which was membranous glomerulonephritis and the others membranoproliferative glomerulonephritis, are reported, emphasizing the clinical course. Two patients had spontaneous remission after seroconversion to anti-HBe positivity, while the third patient was lost to follow-up. We reviewed the management of HBV-associated glomerulonephritis and concluded that immunosuppressive drugs should be avoided since spontaneous remission can be expected in these types of glomerulopathies. PMID- 9223923 TI - Hypophosphatemic vitamin-D resistant rickets associated with epidermal nevus syndrome. A case report. AB - Epidermal nevus syndrome is characterized by congenital anomalies affecting multiple body systems, especially the skin, skeleton and central nervous system. A form of rickets/osteomalacia that is markedly resistant to treatment with vitamin D has been reported in children with this syndrome. We report the clinical and laboratory observations in a child with epidermal nevi and severe hypophosphatemic rickets/osteomalacia. PMID- 9223924 TI - Compound heterozygosity for hemoglobin Knossos [alpha 2 beta 2 27 (B9) Ala-Ser] and IVS I-1 mutation. AB - A three-year-old female with compound heterozygosity for Hb Knossos and IVS-I-1 mutation is presented. On physical examination she had no abnormality except for pallor. Hb was 6.9 g/dl, MCV 61 fl, Hb A2 2% and Hb F 38.5%. Acrylamidegel electrophoresis at a pH of 6 revealed the presence of Hb Knossos in the child and her father. DNA studies revealed that the child was compound heterozygous for Hb Knossos and the IVS I-1 mutation. When the clinical expression of this combination in a previously reported patient with Hb Knossos/FSC8 mutation is compared, it is shown that the newly presented patient has a more severe condition, indicating that the mutations in the trans of Hb Knossos may play a role in the phenotypical expression of the disease. PMID- 9223925 TI - High-dose methylprednisolone-induced dramatic maturation of granulocytes in idiopathic chronic neutropenia. AB - Idiopathic chronic neutropenia is characterized by a variable pattern of age at the onset of neutropenia, neutrophil counts, maturational arrest at the promyelocyte/myelocyte stage in the bone marrow, and recurrent pyogenic infections without any underlying disease or drug administration. There have been reports of lack of response to conventional-dose steroid therapy, but in recent studies it has been shown that high-dose corticosteroids are effective in several neutropenic states. Here we report an apparent response to high-dose methylprednisolone in a patient with idiopathic chronic neutropenia. PMID- 9223926 TI - Intracerebral hemorrhage: a rare late manifestation of vitamin-K deficiency in a breastfed infant. A case report. AB - Late hemorrhagic disease of the newborn (HDN) is a rare complication of vitamin-K deficiency and is especially associated with intracranial hemorrhage. It may also occur in infants who received vitamin-K prophylaxis at birth. Here, we reported a case of late HDN with frontal lobe hemorrhage due to vitamin-K deficiency. This form of intracranial hemorrhage of late HDN has been reported in the literature very rarely. We conclude that the efficiency of single-dose vitamin-K prophylaxis should be revaluated. PMID- 9223927 TI - Autoimmune polyglandular syndrome type I. A case report. AB - Autoimmune polyglandular syndrome (APS) type I is a disorder that consists of three primary diseases: hypoparathyroidism (HPT), adrenocortical insufficiency (ACI) and chronic mucocutaneous candidiasis. Several other disorders may be associated. The diagnosis of APS type I was made in a 16-year-old patient with HPT, Hashimato's thyroiditis and ACI in our department. She has been observed for more than four years for other possible endocrine and non-endocrine disorders. PMID- 9223928 TI - A case of juvenile ankylosing spondylitis and Crohn's disease. AB - Juvenile ankylosing spondylitis (JAS) is a chronic inflammatory arthritis of the peripheral and axial skeleton, frequently accompanied by enthesitis. About four percent of patients with JAS have ulcerative colitis or Crohn's disease. Crohn's disease is the more common of the two and is diagnosed in 26 percent of patients with chronic spondyloarthropathy. In this paper, a 14-year-old male patient is presented as a typical case of juvenile ankylosing spondylitis and Crohn's disease with low back pain, morning stiffness, limited motion in anterior and lateral flexion and extension, left sacroiliitis, ankylosis in the apophyseal joints of the lumbar vertebrae, abdominal pain, bloody diarrhea, characteristic histopathologic changes of colonic involvement such as lymphoid follicles, fissures, submucosal polymorphonuclear cell infiltration and definite ganglion cells. The current therapy with mesalazin, having fewer side effects than sulfosalazin, and its applicability in combination with naproxen sodium is also discussed. PMID- 9223929 TI - Diabetes insipidus in a pediatric patient with systemic lupus erythematosus. A case report. AB - In systemic lupus erythematosus (SLE), the central nervous system (CNS) produces numerous and varying clinical disorders. A 14-year-old girl with the features of cerebral involvement in SLE is presented. The development of diabetes insipidus (DI), which responded to desmopressin, was an interesting finding in this case. Therapy consisting of high-dose methylprednisolone combined with cyclophosphamide was effective in improving the symptoms as well as the DI. PMID- 9223930 TI - Congenital tracheoesophageal fistula without atresia: an incidental finding. AB - A four-year-old boy who had a long history of upper respiratory tract infections and growth retardation was admitted because of recurrent abdominal pain. During upper gastrointestinal series to search for a gastric or duodenal ulcer, the examiner noticed a minute amount of contrast medium within the trachea. Repeat esophagography on an angiographic table led to the correct diagnosis of a congenital H-type fistula. The patient did not have the classical symptoms of a history of choking and cyanosis after feeding during infancy or recurrent lower respiratory tract infections. The only finding consistent with a fistula was growth retardation, and the diagnosis was established incidentally during a work up for abdominal pain. PMID- 9223931 TI - Global AIDS surveillance. PMID- 9223932 TI - The current global situation of the HIV/AIDS pandemic. PMID- 9223933 TI - Education shifts in the 105th Congress. PMID- 9223934 TI - HCFA issues 1997 anesthesia Medicare conversion factors: significant increase in anesthesia services approved. PMID- 9223935 TI - The future of education and accreditation. AB - This information was originally presented during the AANA Annual Meeting in August 1996. Accreditors and educators are urged to plan for the future and maintain quality education by anticipating changes in the work and educational environments. Successful adaptation to change will be critical to the future of education and accreditation. A number of factors serving as catalysts to change are described. PMID- 9223936 TI - Nondisposable sphygmomanometer cuffs harbor frequent bacterial colonization and significant contamination by organic and inorganic matter. PMID- 9223937 TI - Res ipsa loquitur: dental damage during anesthesia. PMID- 9223938 TI - HELLP syndrome (hemolysis, elevated liver enzymes, and low platelets) pathophysiology and anesthetic considerations. AB - HELLP syndrome in the parturient (hemolysis, elevated liver enzymes, and low platelet count) is associated with poor maternal and fetal outcomes. Maternal mortality has been estimated to be as high as 24%. Patients with HELLP syndrome are also at greater risk of pulmonary edema, adult respiratory distress syndrome, abruptio placentae, disseminated intravascular coagulation, ruptured liver hematomas, and acute renal failure. Perinatal mortality is equally high, ranging from 79 to 367 per 1,000 live births, and neonatal complications correlate with the severity of maternal disease. Many clinicians view HELLP syndrome as an entity of preeclampsia, and because of varied symptomatology, the initial diagnosis may be obscured. Prodromal signs include: (1) weakness and fatigue, (2) nausea and vomiting, (3) right upper quadrant and/or epigastric pain, (4) headache, (5) changes in vision, (6) increased tendency to bleed from minor trauma, (7) jaundice, (8) diarrhea, and (9) shoulder or neck pain. Before delivery, aggressive obstetric management is directed toward stabilization of the affected organ systems, if possible, and timely interruption of the pregnancy in the early phase of the accelerated disease progression. Definitive therapy is delivery. Parturients with HELLP syndrome are often critically ill; their infants are frequently premature and their conditions are compromised. Management criteria should include a multidisciplinary approach in a tertiary care center. Obstetric anesthesia personnel should perform a thorough preanesthetic evaluation and be familiar with the pathophysiologic changes of this syndrome. Determining the anesthetic of choice depends on the patient's condition, fetal well-being, and the urgency of the situation. In the presence of severe coagulopathy, regional anesthesia is contraindicated. PMID- 9223939 TI - The expanding role of the clinical coordinator. AB - This article describes the role of the clinical coordinator in nurse anesthesia educational programs, a role which has greatly expanded as programs shift to university-based frameworks with multiple clinical sites. The role of the clinical coordinator has become vital in the success of schools of nurse anesthesia based in the university/college setting. The significance and functional responsibilities of this role need to be broadened by making the clinical coordinator a university appointed representative. The expanded role provides for university representation at each site, improves the lines of communication between the university and each site, and allows for regular visits to each clinical site. Hopefully, program directors, faculty members, and students will benefit from this innovative attempt at bridging the gap between the university and the associated clinical facilities. Regional coordinators also play a vital role in coordinating student functions between various institutions and the clinical coordinators in each of these institutions, as well as representing the university/school of nurse anesthesia interest and needs in these settings. PMID- 9223940 TI - Perioperative pulmonary tumor embolus: a case report. AB - A 64-year-old white female underwent an elective exploratory laparotomy, right radical nephrectomy, and excision of a vena cava tumor for renal cell carcinoma. Preoperatively, the patient was diagnosed with a right kidney mass that was invading the right renal artery and vena cava. The patient had received a general endotracheal anesthetic consisting of O2, N2O, forane, fentanyl, pentothal, vecuronium, and hydromorphone. The right nephrectomy was completed by the urologist without incident. Following the nephrectomy, the right renal artery and vein were dissected, and the vena cava was exposed. When a temporary clamp was placed on the vena cava, there was a sudden onset of tachycardia, right bundle branch block with (transient) inversion of the QRS complex, hypotension, decreased end-tidal CO2 and decreased O2 saturation, and increased pulmonary inspiratory pressures that did not resolve when the clamp was removed. The diagnosis of tumor embolus was made and the incision was closed. As arrangements were made to transfer the patient to a nearby hospital with facilities to perform cardiopulmonary bypass and pulmonary embolectomy, the patient was supported with 100% oxygen, intermittent boluses of epinephrine, sodium bicarbonate, and titrated inotropic infusions (dopamine, dobutamine, and epinephrine). Successful embolectomy was accomplished. The patient was discharged on postoperative day 9 in good condition. PMID- 9223941 TI - Inhaled nitric oxide as a selective pulmonary vasodilator in clinical anesthesia. AB - Inhaled nitric oxide (NO) is a selective pulmonary vasodilator in adult and pediatric patients. Inhaled NO diffuses into the pulmonary vascular smooth muscle where it results in vasodilation via stimulation of guanylyl cyclase. Systemic hemodynamics are not altered because inhaled NO is rapidly inactivated by hemoglobin. Oxygenation is also increased in certain patients as inhaled NO only vasodilates those segments of the pulmonary vasculature which are ventilated. There is growing evidence that inhaled NO may be a useful therapeutic agent in the treatment of pulmonary hypertension and hypoxemia from a variety of causes. Areas of greatest interest to anesthesia and critical care personnel may involve treatment of persistent pulmonary hypertension of the newborn (PPHN), adult respiratory distress syndrome (ARDS), and postoperative pulmonary hypertension secondary to cardiac disease. The potential toxicity of inhaled NO, particularly on immature and developing lungs, must be considered. While inhaled NO exerts acute beneficial effects, it is unclear if there are long-term benefits. Multicenter trials are currently underway to determine if inhaled NO decreases mortality from PPHN or decreases morbidity associated with ARDS. PMID- 9223942 TI - Hyperthermia during repair of pectus excavatum. AB - We have observed an elevation of body temperature during surgical repair of pectus excavatum. To document this phenomenon and attempt to prevent it, we undertook a combination retrospective-prospective study. The retrospective arm included an analysis of the anesthetic records of patients undergoing repair of pectus excavatum during the past 5 years and included 22 boys and 3 girls. Body temperature increased in all 25 patients, from a starting temperature of 36.1 degrees C +/- 0.5 degree C to 38.0 degrees C +/- 0.6 degree C. The maximum temperature exceeded 38 degrees C in 12 patients and 39 degrees C in 3 patients. An additional retrospective review of the anesthetic records of 15 children undergoing another type of thoracic procedure (thoracotomy) for noninfectious problems revealed only a modest, statistically nonsignificant rise in temperature from a starting point of 35.9 degrees C +/- 0.7 degree C to a maximum of 36.3 degrees C +/- 1.2 degrees C. The maximum temperature was greater than 38 degrees C in one of these 15 patients. The prospective arm of the study included a standardized anesthetic technique in 10 patients. Neither a heated humidifier nor a warming blanket were used, and the ambient temperature of the room was maintained at 70 degrees F. Core body temperature increased from a starting temperature of 36.1 degrees C +/- 0.6 degree C to 36.8 degrees C +/- 0.8 degree C. A significant elevation of body temperature occurs in children during repair of pectus excavatum that may be avoided by eliminating the use of exogenous methods to prevent hypothermia (e.g., heated humidifier, warming blanket). PMID- 9223943 TI - AANA Journal course: update for nurse anesthetists--thoracic trauma. AB - The lungs are remarkably resilient organs and injuries to the lungs, whether from blunt or penetrating trauma, are not often the source of mortality. Instead, associated injuries to the heart, great vessels, chest wall, and other critical structures account for most morbidity and mortality associated with thoracic trauma. Hypovolemic shock, impending cardiac failure, neurological injury, and compromised respiratory function may all be present in a single individual with thoracic trauma whose injuries require immediate surgery. Anesthesia care for the thoracic trauma patient often involves continued resuscitation to combat shock, as well as the careful administration of anesthetic agents. The choice of anesthetic agents must consider the patient's hemodynamic status, the potential for airway problems, associated injuries, and medical history. PMID- 9223944 TI - Labor intrathecal analgesia. PMID- 9223945 TI - Short stay. The art of legislating quality and economy. PMID- 9223946 TI - Speaking up for baby. The case for individualized neonatal discharge plans. PMID- 9223947 TI - Hormonic convergence. Finding balance through hormone replacement therapy. AB - Most women today live longer than 80 years, and with menopause beginning anywhere from a woman's late 30 s to early 50 s, most women can expect to live for 30 postmenopausal years or more without the beneficial and protective affects of estrogen. PMID- 9223948 TI - Is your future in staff nursing? PMID- 9223949 TI - Who has the right to say "good job?". PMID- 9223950 TI - Medicare, managed care top congress' agenda. PMID- 9223951 TI - Is it time to upgrade my credentials? PMID- 9223952 TI - Holding nurses accountable. AB - In today's changing health care system, the responsibilities of and the accountability for professional practice has not changed. Regardless of the health care setting, professional nurses are morally, ethically, and legally accountable for their nursing judgments and actions. PMID- 9223953 TI - Women as health care consumers. PMID- 9223954 TI - RCN provides framework for all-graduate profession. PMID- 9223955 TI - Advanced nursing practice: role or concept? PMID- 9223956 TI - Nursing care of the mechanically ventilated patient in itu: 2. AB - This article, the second in a two-part series, outlines the role of the nurse in assessing and implementing aspects of care of the mechanically ventilated patient in the intensive care unit, including sedation, communication, safety, nutrition, and the needs of patients' visitors. The ultimate goal (Where possible) is to help the patient manage without the ventilator. This process is referred to as 'weaning' and often involves the nurse participating in intricate patient assessment and the implementation of specialized care. The need for a multidisciplinary approach to patient care is emphasized. PMID- 9223957 TI - A review of beta-haemolytic streptococcal infection in babies. AB - The incidence of beta-haemolytic group B streptococcal infection in babies at or shortly after birth appears to have risen during the last 20 years. About 30% of women could be colonized at any one time, and because of the transient nature of this organism research has failed to find a way of identifying women who will be colonized at the end of pregnancy. The baby is at risk of early-onset disease from mother-to-baby transmission during the birth process. If identified early enough, those mothers who present a risk can be treated in time to protect the baby during the birth. Late-onset disease can be the result of cross-infection from carers, and the education of midwives, nurses and mothers could reduce the risk. Education could also aid identification of the early signs of disease, and therefore increase the likelihood of a successful outcome of treatment. PMID- 9223958 TI - The role of Sterigel hydrogel wound dressing in wound debridement. AB - An increasing number of hydrogel dressings have become available to the practitioner over the last few years. Sterigel (Seton Healthcare) is the latest to be launched. This dressing is able to donate large quantities of fluid and thereby speed up the debridement of slough and necrotic tissue. PMID- 9223959 TI - Management of the patient with minor traumatic brain injury. AB - Staff in the accident and emergency department and critical care unit are often confronted with patients who have sustained life-threatening injuries. In such situations the symptoms of a minor traumatic brain injury (MTBI) may be missed. It is important that, when the physical symptoms begin to subside, nurses are aware of the symptoms of MTBI and respond accordingly. A post-concussion syndrome may develop 2 weeks to 2 months after a MTBI. This syndrome can affect the patient's ability to perform the usual activities of living. This article defines MTBI, describes the symptoms encountered in patients with MTBI, and outlines the pathophysiology, clinical findings, treatment and nursing interventions. PMID- 9223960 TI - The use of restraint in the care of elderly patients. AB - The use of physical and chemical restraint in elderly care settings to restrict mobility and control behaviour is not a new concept. However, until the 1980s, it was an area that was neglected in nursing research. In 1989, Evans and Strumpf carried out an extensive literature review on restraint in an attempt to consolidate the state of knowledge regarding its use in care of the elderly settings. This article aims to evaluate how far the discussion surrounding restraint has progressed since that time and in what direction. It also reviews the extent of restraint use and rationale for its application, the consequences of restraint and progress in developing alternatives. PMID- 9223961 TI - Medical investigations. 3: Percutaneous liver biopsy. PMID- 9223962 TI - The dilemma of incorporating research into clinical practice. AB - Despite pressure to move towards evidence-based practice within the health service, a considerable proportion of nursing intervention continues to be founded on time-honoured tradition and historical precedent. Most explanations for this research/practice hiatus have focused on organizational problems such as the lack of appropriate skills training, inadequate time and limited resources. This article suggests that the problem may also relate to psychological resistance from nurses, especially at the level of professional stereotyping, a lack of confidence and entrenched gender stereotypes, all of which predispose nurses to maintaining the traditional basis of clinical practice. If nursing is to become a more scientific, clinical service then an open-minded and flexible approach which takes account of the relevant psychological factors is required. PMID- 9223963 TI - Nurses as hotel hostesses: a future path in nursing? PMID- 9223964 TI - Guidelines for nursing care of the patient receiving sedation and analgesia in the gastrointestinal endoscopy setting. Society of Gastroenterology Nurses and Associates, Inc. PMID- 9223965 TI - Standards for infection control and reprocessing of flexible gastrointestinal endoscopes. Society of Gastroenterology Nurses and Associates, Inc. PMID- 9223966 TI - Changes in healthcare system mean opportunities for nurses. PMID- 9223967 TI - Developing a conscious sedation program: from policy development through quality improvement. AB - Conscious sedation (CS) has rapidly become a popular method of anesthesia, administered to decrease the pain and anxiety associated with operative or invasive procedures while avoiding the potential complications of general anesthesia. Intravenous conscious sedation (IVCS) is most frequently administered, but the patient can also receive CS by oral, intramuscular, or rectal routes. Although the level of sedation using CS is not as deep as general anesthesia, the patient is potentially at risk for respiratory depression. CS provides many benefits. It also places an increased burden on healthcare providers to ensure safe and competent care when this form of anesthesia is used. PMID- 9223968 TI - Maintaining competent practice: information settings for the gastroenterology nurse. AB - Professional standards of practice for the gastroenterology nurse address the nurse's need to maintain and enhance professional competency through knowledge acquisition. With the rapidly increasing knowledge base in the nursing profession, it is increasingly important for the nurse to "keep current." In this article, the authors present information settings that assist the gastroenterology nurse in information retrieval and management. PMID- 9223969 TI - Long-term gastrostomy in children: caregiver coping. AB - As technologic interventions such as long-term gastrostomy in children with severe disability become commonplace, and increasing numbers of medically complex children are cared for in the home, nurses will require specialized knowledge to support families. In this article, the authors document findings from a longitudinal study of families caring for a child with a gastrostomy, specifically addressing the caregiver perspective on how families cope with this challenge. The findings reveal a complex set of effective coping strategies as articulated by family caregivers experienced in the management of long-term gastrostomy. Further, these caregivers reveal insights about gastrostomy's technical, social, and psychologic implications for both themselves and their children. In documenting an insider perspective on this phenomenon, this study provides nurses with a comprehensive orientation to the ways in which healthcare professionals can better support family caregiver coping. PMID- 9223970 TI - A licensed practical nurse/licensed vocational nurse's (LPN/LVN) guide to the changing healthcare system. AB - The purpose of this article is to help licensed practical nurses/licensed vocational nurses (LPN/LVN) become aware of their role in the changing healthcare system. There is no better time than the present to begin thinking of how to survive in the current atmosphere of change and uncertainty. The challenge for the LPN/LVN is to define a framework for practice in the new system. A clear vision is needed for tomorrow, a personal vision that each individual must define and work toward. PMID- 9223971 TI - Advanced practice nursing: merging the clinical nurse specialist and nurse practitioner roles. AB - Advanced practice nursing has taken a preeminent role in the recent reform of health care by fostering a perception of leadership, accountability, and responsibility in areas involving access to care, quality of care, and the cost of care. Despite this opportunity for recognition and advancement in the current healthcare system, variations in educational preparation, public confusion about the various roles and titles within advanced practice nursing, and differing professional certifications plague the opportunity for an elevated status of advanced practice nursing. In this article, the author addresses the issues related to merging the clinical nurse specialist and nurse practitioner roles. A review of the historical development of advanced practice nursing along with current practice issues and future needs is also presented. PMID- 9223972 TI - My introduction to metallic esophageal stents with case reviews. PMID- 9223973 TI - Common vitamin B complex agents and folic acid: Part III. Niacin (nicotinic acid) (Niacinamide, Nia-Bid, Niac, Niacels, Nico-400, Nicobid, Nicolar, Nicotinex, Slo Niacin, Span-Niacin, Tega-Span). PMID- 9223974 TI - Nursing in the big picture. PMID- 9223978 TI - Beating the odds. Helping problem gamblers. PMID- 9223979 TI - Beyond our borders. How to get involved in international development. PMID- 9223980 TI - Pakistan's community health workers. AB - Pakistan's health characteristics are worse than those of other Asian countries at similar stages of development. Its mortality rate for children under five is 139 per 1,000, and its maternal mortality is 60 per 10,000. Malnutrition in women and children is widespread; 50 per cent of children under five are stunted. Pakistan's population growth rate of 3.1 per cent per year is among the highest in Asia. The high population growth rate and poor health status of many people call for extensive health care services, but, unfortunately, health services do not reach most of the people of Pakistan. Partly because the training of doctors and nurses is lengthy and expensive, there is an acute shortage of health care providers, especially women. Although female health professionals are preferred for caring for women, cultural constraints inhibit women from seeking education. Such is the multifaceted dilemma in the provision of primary health care in Pakistan. PMID- 9223981 TI - Occupational health and safety in Canadian health care facilities. AB - Numerous authors have expressed concern about the employee health and safety services offered by health care facilities in Canada. More than perhaps any other type of employer, these facilities are well equipped to provide extensive occupational health programs and should be sensitive to the need, but in reality too few facilities offer too little in the way of services. Compounding the problem are the strains resulting from nationwide cutbacks in health care spending, which have seriously deteriorated the quality of working life in Canadian health care facilities. Occupational health and safety programs are correspondingly more important, but are still too rare. PMID- 9223982 TI - Streamlining discharge planning. PMID- 9223983 TI - Let's celebrate nursing. PMID- 9223984 TI - Just the way it is here in Nicaragua. PMID- 9223985 TI - It's a small world. PMID- 9223986 TI - The measure of success. PMID- 9223987 TI - Home care of the elderly. Oral cancer patient. AB - Elderly patients with oral cancer present many challenges in the home healthcare setting. Timely detection of disease, appropriate and timely referrals, and the adverse effects of therapies, nutrition, and depression are discussed in this article. Using a case study, the author offers suggestions on the coordination of services of dental, medical, and nursing professionals and management of adverse dental effects. PMID- 9223988 TI - Clinical laboratory improvement amendment. PMID- 9223989 TI - Violence. Prevention in the home health setting. AB - Home care workers face an increasing risk for workplace violence. A proactive preventive approach is presented that suggests strategies to use during the previsit phase, the visit experience, and on an ongoing basis. PMID- 9223990 TI - Managing cancer pain crisis effectively in the home. PMID- 9223991 TI - Psychosocial interventions for patients with chronic obstructive pulmonary disease. AB - Depression, hopelessness, and pessimism are common behaviors seen in patients with chronic obstructive pulmonary disease (COPD). Healthcare personnel have traditionally focused on therapies to treat the physiologic effects of this debilitating disease. The psychologic problems associated with COPD that can affect a patient's personal and social quality of life are not often addressed. This article reviews current research on these problems and suggests nursing interventions that can assist in meeting patient-centered outcomes. PMID- 9223992 TI - Key moral principles applied to managed care. AB - Although MCOs pose many challenges to the ethics of healthcare delivery, they also have the potential to minimize some previously existing unethical practices. The focus on maximum quality and cost-effectiveness places more demands on the provider with increased accountability. In the old unstructured, noncompetitive healthcare environment, providers were not as accountable to the patient for providing the best quality care at the best price. As home care agencies integrate with managed care networks, outcome measurement becomes increasingly important (National Association for Home Care, 1996). In the managed, integrated market, competition is strong and quality becomes a benchmark when alliances and mergers occur. Home care is in a prime position to be a major player in this new healthcare system and continue to expand services in a capitated environment while being challenged to uphold the ethical practice philosophy outlined in the basic moral principles discussed in this article. PMID- 9223993 TI - A Mom & Me program. AB - For the postpartum patient and her newborn, the hospitalization period continues to decrease while educational needs are unchanged. This leads to high stress and a limited ability for patients to absorb needed information for safe care after discharge. The program described below shows how one home care agency combined the skills of the in-hospital nursing staff and home care staff to develop an in home educational program for this population. PMID- 9223994 TI - Psychiatric symptoms in a community-based medically ill population. AB - Twenty-five home health nurses used the Psychiatric Symptom Assessment Scale to assess the prevalence of psychiatric symptoms in a convenience sample of 176 medically ill clients in their caseloads. Psychiatric symptoms were observed in 74% of the clients, with somatic concerns, anxiety statements, depressive mood, tension, and helpless/hopeless statements being the most prevalent symptoms. Although most symptoms were rated as mild or moderate, several clients had severe symptoms. Implications for nursing care are discussed. PMID- 9223995 TI - Never lose heart? PMID- 9223996 TI - Managing academic development. PMID- 9223997 TI - The language of management: an enduring challenge. AB - The language used in health care organizations has a profound effect on the way people conduct their work. The aim of this paper is to highlight and examine the significance of language in health care organizations, and raise awareness of its relevance to nurse managers. This is achieved by exploring three main themes. The contrasting views of health care, reflected in the language of management and nursing; the management of rhetoric; and researching language. It is argued that the material effects of language need to be recognized by nurse managers, if they are to fulfil their role effectively, and that language can also be used as a resource in the pursuit of this goal. Recommendations are made concerning the need for nurse managers to become sensitive to the varied dimensions of language and a means of initiating this process is suggested. PMID- 9223998 TI - Changing childbirth--the midwifery managers' tale. AB - This paper concerns Maternity Services. It begins by describing the development of the current policy in Britain which culminated in the report Changing Childbirth. It proceeds to describe a qualitative investigation in which a group of Heads of Midwifery Service in one health region were interviewed concerning their views about the basic recommendations of the report and their experiences concerning the feasibility of its implementation by 1998. On the basis of its evidence, the study concludes the policy is failing to be fully realized due to both a lack of earmarked resources and confused objectives which are not fully accepted by professionals and are not understood by service users. The findings demonstrate that the Heads of Midwifery believe that what was proposed as a bold and innovative policy has become no more than lukewarm advice. The conduct of the study is described in detail to illustrate how operational managers can research the ways in which policies are influencing service delivery in their field of practice. PMID- 9223999 TI - Contracting in the National Health Service (NHS): recognizing the need for co operation. AB - Within the reorganized National Health Service hierarchical relationships between Health Authorities and Trusts have been replaced by functional differentiation. However, differentiation of function cannot be seen as an end in itself and management of the relationship between purchasers and providers must include managing the differentiation as well as the function. This paper suggests that collaborative and administrative activities have a distinct role to play in health service management. The paper suggests that in health service management market strategies are likely to dominate in relation to resource allocation activities. The paper also argues that administrative strategies are likely to be necessary within the internal market system--to bridge the gap resulting from the differentiation of function. PMID- 9224001 TI - Job diagnostic surveys of paediatric nursing: an evaluative tool. AB - Two distinct trends can be identified in the context within which nursing care is planned and delivered. One is the continuous pressure to find ways of increasing efficiency and cost-effectiveness. The second is the widespread expectation that public services in general, and health services in particular, should be monitored and evaluated. In these circumstances, nurses and their managers need a range of evaluative tools so that changes in the organization of nursing care can be evaluated. Hackman and Oldham's 'Job Diagnostic Survey' (JDS) approach was tested in a Paediatric Unit in which aspects of primary nursing were being introduced. The paper outlines the JDS approach in the Unit in question and offers an assessment of the value of the JDS as an evaluative tool. PMID- 9224000 TI - The role of clinical directorate managers in facilitating evidence-based practice: a report of an exploratory study. AB - This paper presents an exploration of the role of clinical directorate managers in facilitating evidence-based practice. It reports on a small qualitative study which explored the perceptions held by directorate managers regarding their role in facilitating evidence-based practice for their nursing teams. Insights were sought into the nature of the overt and covert behaviours which the managers used in encouraging nurses to base their practices on research evidence. The sample consisted of 10 directorate managers from two teaching hospitals in one Trust, whose characteristics were not atypical from other directorate managers. Data were collected using semi-structured, tape-recorded interviews. Interviews focused on manager's backgrounds, professional responsibilities and practices, personal research agendas and socio-political influences. Findings revealed some constraining influences on the development of evidence-based practice, such as the Clinical Managers' budget allocations, and the policies and goals of the Trust. However, in respect of the managers' own practices, whilst some facilitative behaviours were identified, the data suggested that clinical managers were behaving in a manner which actually inhibited the development of evidence-based nursing practice. It would appear that in the post-reform National Health Service (NHS) clinical managers find themselves in a difficult position which gives rise to a practical failure to use their position and organizational authority to influence the utilization of research in practice. The implications of these findings for an evidence-based health service are discussed. PMID- 9224002 TI - Expert views on clinical supervision: a study based on interviews. AB - Clinical supervision is a didactic process of the purpose of human development and maturity. The aim of this study is to analyse views on clinical supervision held by a number of experts, and to reflect on the effects and value of clinical supervision in relation to public health. Data were collected by interviews and analysed in accordance with the grounded theory construction model. The results showed that clinical supervision is an integration process guiding a person from 'novice to expert' by establishing a relationship of trust between supervisor and supervisee. This study indicates that implementation of systematic clinical supervision may positively affect quality of care, and patients' recovery, create improved feeling of confidence in one's work, and prevent burnout among staff. The negative aspects, as reported, were the possibility of high 'opportunity costs', e.g. the time loss for patient care by those participating in organized systematic supervision. On the other hand, clinical supervision contributes towards more efficient use of resources and hence avoids unnecessary costs. However, neither of these aspects were further elaborated on by the experts but clearly indicate an important field for further research. PMID- 9224003 TI - Towards a framework for clarifying psychiatric nursing roles. AB - Psychiatric nurses function in many varied settings of practice, against the backdrop of continuous change, in the organization of health care delivery. Understandably, psychiatric nursing practice roles are not well-defined despite the availability of a range of theories. A framework for brokering between theory and practice is proposed. Elements of role adequacy, role legitimacy and role support are related to 'theory of' and 'theory for' psychiatric nursing. When treated as dynamic in relation to one another, these elements can form a framework for clarifying practice roles. PMID- 9224004 TI - Dual diagnosis of severe mental health problems and substance abuse/dependence: a major priority for mental health nursing. AB - It is now established that very significant numbers of people with severe mental illness abuse or depend on drugs and/or alcohol. This combination (Dual Diagnosis) leads to increased rates of violence and service use, a reduction in adherence to treatment regimes, an increase in susceptibility to human immunodeficiency virus (HIV) infection and is now found in in-patient populations. Because of their vulnerability to accidents and physical illnesses, dual diagnosis patients are found increasingly in accident and emergency departments, general medical wards and primary care settings. For this reason nurses and other health professionals working in general hospitals should be as aware as their mental health colleagues of the specific needs of this population. There are some excellent models of service organization and training for dealing with dual diagnoses populations in some parts of the USA. However, there is little such development in the UK. There are clear pathways to be followed, but the need for action is urgent. PMID- 9224005 TI - Children of the mentally ill: a qualitative focus group approach. AB - Children of the mentally ill constitute a group neglected by mental health care providers. Increased rates of psychopathology, impaired attention processes, disturbances in interpersonal relationships, and reduced overall adaptive functioning are reported as significant outcomes for offspring of parents with a mood disorder. While epidemiological studies underscoring the risks from a hereditary standpoint are many, there are few studies examining the subjective experience of living with a depressed parent. Findings from this pilot study elucidate the subjective experience of preadolescents/adolescents living with an affectively ill parent, applying a qualitative focus group design. Videotaped sessions were analysed using methods consistent with qualitative research. 'Struggle to understand the illness', 'managing the illness', 'recognizing the signs', and 'impact of parent's hospitalization' emerged as central themes, capturing the essence of participants' experiences. The first two themes were further divided into subthemes. Findings illuminate the need to broaden nursing interventions and research, to include family perspectives, particularly when parental mental illness is a factor. PMID- 9224006 TI - Evaluation of a low-threshold clinic for opiate-dependent drug users. AB - This study reports on the development and outcome of a Low Threshold Clinic (LTC) for opiate-dependent drug users. The service originated as a nursing initiative within an inner city Drug Dependency Centre (DDC) and its rationale and treatment approach are explored in relation to the literature and local circumstances. Client baseline and outcome data were systematically gathered to assess service uptake and service efficacy in terms of client outcome. Data are presented for the first two years of operation during which a total of 59 clients entered the LTC. The sociodemographic characteristics and patterns of drug use among this group suggest the service was successful in targeting clients who previously failed to enter traditional treatment programmes despite initial referral to the DDC. Outcome data indicate a tendency for clients to inject less frequently, engage in less criminal activity and, by 12 months, to reduce their dose of prescribed methadone whilst attending the LTC. Tentative conclusions are drawn concerning the value of this service for 'hard-to-reach' drug users and those who may be at a precontemplation stage of change. Recommendations are made for a more comprehensive evaluative study that involves comparison with other treatment approaches. PMID- 9224007 TI - Generating ideas for teaching mental health nursing. AB - University teachers who prepare mental health nurses require a variety of teaching techniques to encourage students to engage with the subject matter and to prepare for unknown future problems and dilemmas that they must resolve. Mental health nursing requires the development of specific discipline knowledge, intellectual skills, such as the ability to think laterally and make ethical decisions, and personal attributes, such as maturity and tolerance, as well as practical experience in collaborative care. This article offers a number of strategies that teachers may use in their classes to promote the development of these mental health nursing skills. Other teachers might use these examples to trigger the development of their own repertoire of teaching skills for beginning mental health nurses. PMID- 9224008 TI - Nurses and neuroleptic medication: applying theory to a working relationship with clients and their families. AB - This paper presents a model for the nursing management of the effects of neuroleptic or antipsychotic medication. The model addresses the contemporary issues facing nurses, their clients and families, while at the same time is congruent with the continua of effects of these medications. For many, nonadherence is directly related to undesired side effects, while for others the reasons may be multidimensional. The proposed model addresses the notion of advocacy and empowerment in the light of interventions that relate to the undesired-desired effects continuum. Underpinning this is the notion of the phases of the nurse-client relationship, which were advocated by Peplau (1952) and have since offered a framework for nursing intervention. While developing the phases of this relationship, nurses can help the client and family by assuming various roles that can enhance the relationship. It is suggested that each of these roles has an element of advocacy and an element of empowerment. It is part of the nurses' professional responsibility to help the client and their family to deal with the issues surrounding medication nonadherence. PMID- 9224009 TI - Mentoring state hospital nurses to write. AB - There is a shortage of clinical nurses, particularly in the area of psychiatric nursing, who feel competent and comfortable writing for publication. This is unfortunate since clinical nurses have valuable experiences to share, to promote and advance the nursing profession. This manuscript describes the work of one Director of Nursing at a state hospital who mentored the nursing staff to write for publication. Over a period of five years some 28 articles have been published by nurses at this hospital. This success in publication has led to an increase in self-esteem among nurses, an increased willingness to share their writing experiences with others, and increased numbers of workshop presentations on clinical subjects. This article describes how the mentoring process can be used to promote authorship, and specifically, how it was used at this one hospital. PMID- 9224010 TI - Liaison psychiatric nursing in an inner city accident and emergency department. AB - The Mental Health Nursing Review Team recommend research to examine the potential of liaison mental health nursing. This paper describes a study into the work of two emergency psychiatric nurses (EPN) attached to a central London accident and emergency (A&E) department. A semi-structured interview schedule was employed to generate data on role development and purpose, and a retrospective survey of clinical activity was carried out over a 94-day period. Specific attributes and working practices, which promoted integration within the A&E department, are identified as are the reasons for referring patients to the EPNs, patients' primary problems on assessment, assessment outcome and the staff involved in each assessment. Findings are considered in relation to the various models of psychiatric consultation and liaison found in the literature. The scope and quality of the available data is also assessed and recommendations are made to enable a more robust prospective study to be undertaken. PMID- 9224011 TI - Manipulativeness and dialogue. AB - The notion of manipulativeness is sometimes invoked both with regard to patients or clients and with regard to the professionals who are charged with caring for them. The paper explores that notion and the idea that what makes an action manipulative is not the end at which it is directed but the way in which that end is sought. Open dialogue is characterized and it is suggested that manipulative interactions are ones that depart from open dialogue. Manipulative persons are characterized as having or being biased towards a restricted social repertoire. While some manipulativeness is inevitable, it is suggested that we need to reflect from time to time on whether we have got the balance right. PMID- 9224012 TI - The epidemiology and effects of borderline personality disorder in primary health care. PMID- 9224013 TI - The contribution of nurses, midwives and health visitors to the targeting of health and social need in Northern Ireland. PMID- 9224014 TI - Mental health pathways to care. PMID- 9224016 TI - The future of community psychiatric nursing. PMID- 9224015 TI - The development of a Mental Health Nurse Practitioner service in an acute district hospital. PMID- 9224018 TI - The added value of board certification status. PMID- 9224019 TI - The care and feeding of a Web page: a call for case discussions. PMID- 9224021 TI - Brown recluse spider bite: a case study. AB - The case of a young man with a chronic, resistant, and unpredictable wound is described. The wound is believed to have been caused by the bite of a brown recluse spider. This article presents a brief overview of the potential sequelae of brown recluse spider bite and a pictorial review of this young man's wound during a 22-month period. PMID- 9224022 TI - Nursing wound care survey: sterile and nonsterile glove choice. AB - PURPOSE: The application of sterile and clean procedure to the practice of wound care nursing was examined. DESIGN: This prospective, descriptive study surveyed staff nurses regarding glove use. SUBJECTS AND SETTING: Seven hundred forty-three staff nurses from five health care agencies in the San Francisco Bay Area responded to the survey. INSTRUMENTS: A self-report wound care survey instrument was developed by Nursing Consortium for Research and Practice members from information adapted from the wound care literature. The questionnaire comprised 31 questions and required approximately 10 minutes to complete. METHODS: Nursing Consortium for Research and Practice members obtained approval from their respective institutional human subjects committees and distributed questionnaires among all nurses engaged in direct care. Some agency representatives personally handed the survey instruments to subjects, but most distributed them through their agencies personnel mailing systems. RESULTS: Seven hundred twenty-three (38%) of 1900 questionnaires were completed and returned to the five site coordinators. Differences were found between acute care and home health nurses. Acute care nurses were more likely than home care nurses to use sterile gloves in all wound care situations. CONCLUSION: Greater variation was found with regard to sterile technique in wound care practice than in previously reported studies. Although patient risk factors and wound type significantly influenced the choice of sterile or clean gloves, additional environmental and personal factors exerted considerable influence. These included health care setting, degree of professional education, and nurses' experiential background. Attempts to modify practice through policy change alone may not be sufficient to overcome resistance to change. Instead, it may be necessary for nurses to "unlearn" lessons from basic nursing education before they can adopt to new practices and clinical policies. PMID- 9224020 TI - Surviving redesign: basic concepts of patient-focused care and their application to WOC nursing. AB - The concept of patient-focused care continues to gain popularity in health care delivery throughout the United States. In an effort to compete more effectively in today's market, health care institutions have adopted strategies from industry to control rising costs. WOC nurses in every practice setting are feeling the effects of one such strategy, patient-focused care. The purpose of this article is to describe basic concepts of patient-focused care, a redesign technique that was born out of continuous quality improvement, and to present strategies tailored to strengthen WOC nursing practice in this changing and challenging environment. PMID- 9224025 TI - Notes on methodology. Participation of human subjects in studies. PMID- 9224024 TI - Home-based management of urinary incontinence: a pilot study with both frail and independent elders. AB - OBJECTIVE: The participation of older rural women and their caregivers in a pilot research study on behavioral management interventions for urinary incontinence is described. DESIGN: A quasiexperimental design was used. SETTING AND SUBJECTS: Women 55 years old and older and living in a rural county in North Florida who had episodes of urinary incontinence twice or more per week were included. Outreach was directed at two groups of elders with incontinence, those who were functioning independently and those who were frail and dependent on caregivers for assistance with activities of daily living. METHODS: Behavioral management of continence comprised three techniques for the management of urinary incontinence: self-monitoring, scheduling regimens and pelvic muscle exercise with biofeedback. MAIN OUTCOME MEASURES: The main outcome measures were episodes of urine loss and amount (In grams) of urine loss with time, determined with a weighed pad test. RESULTS: Behavioral management of continence resulted in significant decrease in urinary incontinence; decreases in frequency and volume of urine loss were found among all study participants when data were analyzed. CONCLUSIONS: Although behavioral management of continence was effective in reducing incontinence among independent, community-dwelling elderly women, there was a marked lack of response to this project by frail elders and their caregivers. The same barriers to implementing time-intensive behavioral management interventions among frail elders in long-term care facilities may operate in the home setting. PMID- 9224023 TI - Pregnancy, delivery, and postpartum experiences of fifty-four women with ostomies. AB - OBJECTIVE: The pregnancy, labor, and delivery experiences of women with ostomies are described. DESIGN: Patient survey. SETTINGS AND SUBJECTS: Distribution of surveys to women with these experiences: patients of the Cleveland Clinic Foundation, referrals by networking of ET nurses, and distribution of surveys at a national conference of the United Ostomy Association and through local ostomy associations. INSTRUMENT: A 12-point questionnaire was adopted for each type of diversion: ileostomy, urostomy, continent ileostomy, colostomy, and pelvic reservoir. METHOD: The survey was distributed to attendees at a national United Ostomy Association conference and local United Ostomy Association meetings. Surveys were distributed to other EI nurses in the mideastern United States region and to qualified patients at Cleveland Clinic Foundation. MAIN OUTCOME MEASURE: The subjective experiences of pregnancy, labor and delivery of women with ostomies are described. RESULTS: Seventy-five questionnaires were distributed and 54 women responded. Eight women (15%) had fertility problems. Stoma-related problems during the second or third trimester were reported by 68.5%. Most problems were corrected without medical intervention. CONCLUSIONS: The presence of an ostomy should not be a deterrent to successful pregnancy and delivery. PMID- 9224026 TI - Continence management and wound care for a patient in a residential facility. PMID- 9224027 TI - A legend in ET nursing: a tribute to the memory of Ruth Kules, RN, CETN. PMID- 9224028 TI - The societal burden of incontinence: even greater than we thought. PMID- 9224029 TI - You say sterile and I still say not: a response to my colleague's letter to the editor. PMID- 9224030 TI - Compression therapy and venous ulcerations: another point of view. PMID- 9224031 TI - Future shock: millennial midwifery. PMID- 9224032 TI - Water birth--a passing fad? PMID- 9224033 TI - Aurelia, Cecily & Ann: a brief survey of caesarean section. PMID- 9224034 TI - An upside-down world? PMID- 9224035 TI - The legal aspect of sterilisation. Part I. PMID- 9224036 TI - How will it change your practice? PMID- 9224037 TI - Supervisors and midwives' morale in the NHS. PMID- 9224038 TI - First class delivery? PMID- 9224042 TI - The Iolanthe Midwifery Trust. PMID- 9224043 TI - Thinking about nursing futures. PMID- 9224044 TI - Nursing our narratives: towards a dynamic understanding of nurses in narrative tales. AB - Previous research on the representation of nurses in literature has tended to rely on a 'quasi-scientific' method that ultimately produces catalogues of static images. This paper argues that literary representations of nurses must be analysed in terms of situational context. In order to accomplish this the narratological concepts of the chronotope and the donor are used, resulting in a dynamic and powerful reading of nurses in narrative tales. PMID- 9224045 TI - Tradition and culture in Heidegger's Being and Time. AB - In nursing literature, Heideggerian hermeneutics, as expounded in Being and Time, is taken well near unanimously to be an invitation to explore tradition and culture. Understanding, we are told in the name of Heidegger, is to be found in the realm of common meanings and shared practices. This interpretation of what Heidegger is about in Being and Time is neither unchallengeable nor unchallenged. While a number of scholars can be found to agree with it, there are many others who see it as an utter misreading of Heidegger. In their judgement, it is an interpretation diametrically opposed to what Heidegger sets forth in his treatise. For researchers interested in invoking Heidegger or following a Heideggerian approach, this is a frustrating impasse. The only valid starting point for resolving it, this article suggests, is a close reading of what Heidegger actually says in the pages of Being and Time. PMID- 9224046 TI - Postmodern research: no grounding or privilege, just free-floating trouble making. AB - Postmodernism has been criticized as failing to offer, on the one hand, authoritative explanations for social phenomena that might provide a scientific basis for policy formation or, on the other, the philosophical justification for emancipatory work-its radical scepticism about claims to knowledge leaving its advocates, including many nurses, with little scope to transform oppressive social and political regimes. Various approaches to this important problem have been offered, both philosophical and methodological. Some critical theorists have rejected certain aspects of postmodernism as dangerous and distracting. Some more accommodating solutions are troubled by unacknowledged inconsistencies. Others embrace postmodernism's unavoidable ambiguity (towards the Enlightenment for instance) with a lighter heart. In this paper I will review some of the criticism of postmodernism and some proposed solutions to these problems. Using recent research into the impact of managerialism on nursing within the UK National Health Service as an example and drawing on deconstructive literary theory, I conclude by accepting a rhetorical agonistics of undecidability. I take postmodernism as a mandate for causing trouble for those groups who are currently having their say and whose version of truth and rationality has achieved domination over others. I do not take postmodernism as a place from which to champion the cause or privilege the view of any particular group. PMID- 9224047 TI - Reviewing Foucault: possibilities and problems for nursing and health care. AB - This paper addresses Foucauldian theory and its usefulness to nursing research. It is written in the form of a discussion between the authors on the merits and liabilities of Foucauldian theory as applied to analyses of nursing. As such, it focuses upon some of the more pertinent critiques of both Foucauldian and postmodern theory. By addressing Foucault from two different positions, the discussion seeks to demonstrate the complexity of Foucauldian theory and warns against over-simplification in its application to nursing research. The authors also argue for an awareness of the strengths and weaknesses of any given theoretical stance. PMID- 9224048 TI - Grounded theory method, Part I: Within the five moments of qualitative research. AB - While discussing how the grounded theory method has been situated during the five moments of qualitative research history, a detailed analysis is offered of the differences between the classic mode of the method and Strauss and Corbin's reformulation of the method. Such differences include: philosophical perspectives, paradigm of inquiry, intended product, theoretical underpinnings, procedural steps and claims of rigour. Reasons for Strauss and Corbin's elaboration of the method are suggested, within an historical context. PMID- 9224049 TI - The researcher's personal responses as a source of insight in the research process. AB - Drawing on accounts of the author's personal responses while undertaking a qualitative study on the norms governing the relationship between nurses and mothers, it is argued that such responses, rather than being seen as a source of bias, have the potential to be a source of insight and interpretation in the research. This paper tells the 'inside' story of previously published research that was 'sanitized' by the omission of any reference to the researcher's subjective responses. The recognition of such researcher responses has implications for how research is supervised and presented. PMID- 9224050 TI - Why we should wash our hands of medical soaps. PMID- 9224051 TI - A conversation with Margarete Sandelowski and Philip Darbyshire: issues in qualitative inquiry. Interview by Carolyn Emden. PMID- 9224052 TI - Men in nursing: exploring the male nurse experience. PMID- 9224053 TI - The International Council of Nurses Quadrennial Congress. PMID- 9224054 TI - The curse of history. PMID- 9224055 TI - Incompetent criteria. PMID- 9224056 TI - A guy named Joe. PMID- 9224057 TI - Fear factor. PMID- 9224058 TI - Hungry for knowledge. PMID- 9224059 TI - Teaching the teachers. PMID- 9224060 TI - Refuge reaching out. PMID- 9224061 TI - Healthy choice. PMID- 9224062 TI - The Centre for Palliative Care Education and Research. AB - This article describes the role and function of the Centre for Palliative Care Education and Research at the University of Central England in Birmingham. The authors describe the development of the centre, which is a multidisciplinary venture in collaboration with Cancer Relief Macmillan Fund and palliative care providers throughout the west Midlands. Current educational and research initiatives are identified and some future challenges indicated. PMID- 9224063 TI - Back pain in nursing and associated factors: a study. AB - This article reports on an assessment of back pain and associated factors undertaken with nursing staff on nine NHS hospital wards and two private wards. One hundred and sixty eight nurses completed a confidential, retrospective questionnaire. The results suggest that despite the implementation of the 1992 EC manual handling operations regulations, back pain among nurses still remains a problem and is often due to the cumulative effects of work pressures. There is little evidence that training in manual handling reduces the prevalence of back pain directly, since the factors influencing the occurrence of back pain are complex. PMID- 9224065 TI - Student nurses on the Internet. PMID- 9224064 TI - The psychosocial aspects of visual impairment in diabetes. PMID- 9224066 TI - In our own hands. PMID- 9224067 TI - 'I'd like to hear from you'. PMID- 9224068 TI - A student's support helped me cope. PMID- 9224069 TI - Risk, safety, and the dark side of quality. PMID- 9224070 TI - Treating diarrhoea. PMID- 9224071 TI - Chlamydia pneumoniae and coronary heart disease. PMID- 9224072 TI - Screening for people with a family history of colorectal cancer. PMID- 9224074 TI - Russia fears rapid increase in HIV infection. PMID- 9224073 TI - US attempts to keep Medicare solvent. PMID- 9224075 TI - New Filipino law may harm inner city care. PMID- 9224076 TI - Romania adopts new transplant law. PMID- 9224078 TI - Health authority tries to reclaim 700,000 pounds settlement. PMID- 9224077 TI - Wales is set 20 health targets. PMID- 9224079 TI - Randomised controlled trial of effect of fruit and vegetable consumption on plasma concentrations of lipids and antioxidants. AB - OBJECTIVES: To determine the extent to which plasma antioxidant concentrations in people with habitual low intake of fruit and vegetables respond to increased intakes of these foods. To examine whether advice to increase fruit and vegetables will result in reduction of concentrations of total and low density lipoprotein cholesterol. DESIGN: Randomised controlled trial in which intervention and control groups were followed up for eight weeks. The intervention group was asked to consume eight servings of fruit and vegetables a day. SETTING: Dunedin, New Zealand. SUBJECTS: Eighty seven subjects with normal lipid concentrations who ate three or fewer servings of fruit and vegetables daily. MAIN OUTCOME MEASURES: Plasma concentrations of vitamin C, retinol, alpha and beta carotene, alpha tocopherol, lipids, and lipoproteins. Dietary intake assessed with diet records over four days. RESULTS: The mean plasma vitamin C, alpha carotene, and beta carotene concentrations increased in parallel with increased dietary intake of fruit and vegetables in the intervention group. Concentrations of retinol, alpha tocopherol, lipids, and lipoproteins remained unchanged despite some increase in dietary vitamin E and a small reduction in saturated fat intake. CONCLUSIONS: Following a recommendation to increase fruit and vegetable consumption produces change in plasma concentrations of vitamin C, alpha carotene, and beta carotene likely to reduce incidence of cancer. More specific dietary advice to modify fat intake may be necessary to reduce the risk of cardiovascular disease mediated by lipoprotein and vitamin E. PMID- 9224080 TI - Cohort study of effect of being overweight and change in weight on risk of coronary heart disease in old age. AB - OBJECTIVE: To evaluate risk of late life coronary heart disease associated with being overweight in late middle or old age and to assess whether weight change modifies this risk. DESIGN: Longitudinal study of subjects in the epidemiological follow up study of the national health and nutrition examination survey I. SETTING: United States. SUBJECTS: 621 men and 960 women free of coronary heart disease in 1982-84 (mean age 77 years). MAIN OUTCOME MEASURE: Incidence of coronary heart disease. RESULTS: Body mass index of 27 or more in late middle age was associated with increased risk of coronary heart disease in late life (relative risk = 1.7 (95% confidence interval 1.3 to 2.1)) while body mass index of 27 or more in old age was not (1.1 (0.8 to 1.5)). This difference in risk was due largely to weight loss between middle and old age. Exclusion of those with weight loss of 10% or more increased risk associated with heavier weight in old age (1.4 (1.0 to 1.9)). Thinner older people who lost weight and heavier people who had gained weight showed increased risk of coronary heart disease compared with thinner people with stable weight. CONCLUSIONS: Heavier weight in late middle age was a risk factor for coronary heart disease in late life. Heavier weight in old age was associated with an increased risk once those with substantial weight loss were excluded. The contribution of weight to risk of coronary heart disease in older people may be underestimated if weight history is neglected. PMID- 9224081 TI - Secular trend in the occurrence of asthma among children and young adults: critical appraisal of repeated cross sectional surveys. AB - OBJECTIVES: To review repeated surveys of the rising prevalence of obstructive lung disease among children and young adults and determine whether systematic biases may explain the observed trends. DESIGN: Review of published reports of repeated cross sectional surveys of asthma and wheezing among children and young adults. The repeated surveys used the same sampling frame, the same definition of outcome variables, and equivalent data collection methods. SETTING: Repeated surveys conducted anywhere in the world. SUBJECTS: All repeated surveys whose last set of results were published in 1983 or later. MAIN OUTCOME MEASURES: Lifetime and current prevalences of asthma and current prevalence of wheezing. The absolute increase (yearly percentage) in the prevalences of asthma and wheezing was calculated and compared between studies. RESULTS: 16 repeated surveys fulfilled the inclusion criteria. 12 reported increases in the current prevalence of asthma (from 0.09% to 0.97% a year) and eight reported increases in the current prevalence of wheezing (from 0.14% to 1.24% a year). Changes in labelling are likely to have occurred for the reporting of asthma, and information biases may have occurred for the reporting of wheezing. Only one study reported an increase in an objective measurement. CONCLUSIONS: The evidence for increased prevalences of asthma and wheezing is weak because the measures used are susceptible to systematic errors. Until repeated surveys incorporating more objective data are available no firm conclusions about increases in obstructive lung disease among children and young adults can be drawn. PMID- 9224082 TI - Treatment of herpes simplex gingivostomatitis with aciclovir in children: a randomised double blind placebo controlled study. AB - OBJECTIVES: To examine the efficacy of aciclovir suspension for treating herpetic gingivostomatitis in young children. DESIGN: Randomised double blind placebo controlled study. SETTING: Day care unit of a tertiary paediatric hospital. SUBJECTS: 72 children aged 1-6 years with clinical manifestations of gingivostomatitis lasting less than 72 hours; 61 children with cultures positive for herpes simplex virus finished the study. MAIN OUTCOME MEASURES: Duration of oral lesions, fever, eating and drinking difficulties, and viral shedding. INTERVENTION: Aciclovir suspension 15 mg/kg five times a day for seven days, or placebo. RESULTS: Children receiving aciclovir had oral lesions for a shorter period than children receiving placebo (median 4 v 10 days (difference 6 days, 95% confidence interval 4.0 to 8.0)) and earlier disappearance of the following signs and symptoms: fever (1 v 3 days (2 days, 0.8 to 3.2)); extraoral lesions (lesions around the mouth but outside the oral cavity) (0 v 5.5 days (5.5 days, 1.3 to 4.7)); eating difficulties (4 v 7 days (3 days, 1.31 to 4.69)); and drinking difficulties (3 v 6 days (3 days, 1.1 to 4.9)). Viral shedding was significantly shorter in the group treated with aciclovir (1 v 5 days (4 days, 2.9 to 5.1)). CONCLUSIONS: Oral aciclovir treatment for herpetic gingivostomatitis, started within the first three days of onset, shortens the duration of all clinical manifestations and the infectivity of affected children. Further studies are needed to evaluate the ideal dose and length of treatment. PMID- 9224083 TI - Unsupervised surgical training: questionnaire study. PMID- 9224084 TI - Ocular damage associated with proton pump inhibitors. PMID- 9224085 TI - Recruitment, retention, and time commitment change of general practitioners in England and Wales, 1990-4: a retrospective study. AB - OBJECTIVES: To describe the recruitment and retention of general practitioners and changes in their time commitment from 1 October 1990 to 1 October 1994. DESIGN: Retrospective analysis of yearly data. SETTING: England and Wales. SUBJECTS: General practitioners in unrestricted practice. MAIN OUTCOME MEASURES: Numbers of general practitioners moving into and out of general practice; proportion of general practitioners practising less than full time; proportion of general practitioners having unchanged time commitment over the study period; and proportion of general practitioners leaving general practice in 1991 who were subsequently practising in 1994. RESULTS: Numbers of general practitioners entering general practice (1565 in 1990, 1400 in 1994) fell over the study period as did the numbers leaving general practice (1488 in 1990, 1115 in 1994). The net effect was an increase in both the total and full time equivalent general practitioners practising from 1 October 1990 (26,757 full time equivalents) to 1 October 1994 (27,063 full time equivalents). Numbers of general practitioners practising full time were decreasing whereas part time practice was increasing; women were more likely to practise part time. 35.5% (43/121) of women practising full time and 17.8% (24/135) of men practising full time who left practice in 1991 were practising again in 1994. CONCLUSION: Simply using total numbers of general practitioners or net increase to describe workforce trends masks much movement in and out of general practice and between differing time commitments. Recruitment and retention issues need to be separated if reasonable policies are to be developed to assure the necessary general practitioner workforce for a primary care led NHS. PMID- 9224086 TI - Commentary: there is urgent need to raise recruitment--even to stand still. PMID- 9224087 TI - Cognitive behaviour therapy--clinical applications. PMID- 9224088 TI - ABC of mental health. Community mental health services. PMID- 9224089 TI - The injustice of compensation for victims of medical accidents. PMID- 9224090 TI - Managed care. Origins, principles, and evolution. AB - Managed care has entered the lexicon of healthcare reform, but confusion and ignorance surround its meaning and purpose. It seeks to cut the costs of health care while maintaining its quality, but the evidence that it is able to achieve these aims is mixed. As well as raising awareness and understanding of the issues surrounding managed care, this series considers whether managed care is desirable for the NHS. Developed in the United States as a response to spiralling healthcare costs and dysfunctional fragmented services, managed care is not a discrete activity but a spectrum of activities carried out in a range of organisational settings. Due to its constantly changing nature, managed care is a slippery concept--but all its permutations have in common an attempt to influence and modify the behaviour and practice of doctors and other health professionals towards cost effective care. Whatever potential managed care may hold in this regard, careful appraisal of its implications is essential. PMID- 9224091 TI - Nicotine replacement therapy should be prescribable on NHS. PMID- 9224092 TI - Dietary treatment of active Crohn's disease. Dietary treatment is best for children. PMID- 9224093 TI - Dietary treatment of active Crohn's disease. Diet is the best treatment. PMID- 9224094 TI - Hypothesis that people with coronary heart disease are living longer is supported. PMID- 9224095 TI - Cyclosporin can be used in early rheumatoid arthritis. PMID- 9224096 TI - Senior house officer training. Training must be more structured. PMID- 9224097 TI - Senior house officer training. Improved training may have more to do with money than with new shifts. PMID- 9224098 TI - Senior house officer training. Impact of existing peer review visits needs to be increased. PMID- 9224099 TI - Senior house officer training. When midwives perform obstetric tasks at night, trainees can spend more supervised time in clinics. PMID- 9224100 TI - Blood donation, body iron stores, and risk of myocardial infarction. Confidence intervals and possible selection bias call study results into question. PMID- 9224101 TI - Most British research and development in primary care arises outside rural areas. PMID- 9224102 TI - Chaplains on transplant teams facilitate permission being given for organ donation. PMID- 9224103 TI - Amiodarone pulmonary toxicity. Authors did not emphasise typical radiological and histological features sufficiently. PMID- 9224104 TI - Amiodarone pulmonary toxicity. Amiodarone should be used with caution in patients in intensive care. PMID- 9224105 TI - Systematic reviews provide information not contained in traditional narrative reviews. PMID- 9224106 TI - Does setting good practice standards for research ethics committees increase their legal liability? PMID- 9224107 TI - Mechanism of accountability needs to be developed for changing role of healthcare professionals. PMID- 9224108 TI - Alcohol misuse among doctors. Treatment should be offered. PMID- 9224109 TI - Alcohol misuse among doctors. Eliminating stress is the way forward. PMID- 9224110 TI - Breast cancer screening for younger women is not an efficient use of resources. PMID- 9224111 TI - Vegetarians and vegans may be most at risk from low selenium intakes. PMID- 9224112 TI - Oestrogen receptors and breast cancer. PMID- 9224113 TI - Reforming the New Zealand health reforms. PMID- 9224114 TI - Vitamin E and cardiovascular protection in diabetes. PMID- 9224115 TI - HIV associated tuberculosis. PMID- 9224116 TI - Devolution and the Scottish NHS. PMID- 9224117 TI - US tobacco firms agree to $369bn deal. PMID- 9224118 TI - Health authorities plan legal action against tobacco companies. PMID- 9224120 TI - Russian doctors protest over state of nation's health. PMID- 9224119 TI - South African Truth Commission calls doctors to account for their actions during the apartheid era. PMID- 9224121 TI - UK government to tackle prescription fraud. PMID- 9224122 TI - Colombia is confused over legalisation of euthanasia. PMID- 9224123 TI - Institute for Medical Ethics faces up to flood of dilemmas. PMID- 9224124 TI - Links with leukaemia confirmed for French nuclear plant. PMID- 9224125 TI - The union gets interested in ethics. PMID- 9224126 TI - Corticosteroids in acute traumatic brain injury: systematic review of randomised controlled trials. AB - OBJECTIVE: To quantify the effectiveness and safety of corticosteroids in the treatment of acute traumatic brain injury. DESIGN: Systematic review of randomised controlled trials of corticosteroids in acute traumatic brain injury. Summary odds ratios were estimated as an inverse variance weighted average of the odds ratios for each study. SETTING: Randomised trials available by March 1996. SUBJECTS: The included trials with outcome data comprised 2073 randomised participants. RESULTS: The effect of corticosteroids on the risk of death was reported in 13 included trials. The pooled odds ratio for the 13 trials was 0.91 (95% confidence interval 0.74 to 1.12). Pooled absolute risk reduction was 1.8% ( 2.5% to 5.7%). For the 10 trials that reported death or disability the pooled odds ratio was 0.90 (0.72 to 1.11). For infections of any type the pooled odds ratio was 0.92 (0.69 to 1.23) and for the seven trials reporting gastrointestinal bleeding it was 1.05 (0.44 to 2.52). With only those trials with the best quality of concealment of allocation, the pooled odds ratio estimates for death and death or disability became closer to unity. CONCLUSIONS: This systematic review of randomised controlled trials of corticosteroids in acute traumatic brain injury shows that there remains considerable uncertainty over their effects. Neither moderate benefits nor moderate harmful effects can be excluded. The widely practicable nature of the drugs and the importance of the health problem suggest that large simple trials are feasible and worth while to establish whether there are any benefits from use of corticosteroids in this setting. PMID- 9224127 TI - Prospective study of effect of switching from cigarettes to pipes or cigars on mortality from three smoking related diseases. AB - OBJECTIVE: To estimate the extent to which cigarette smokers who switch to cigars or pipes alter their risk of dying of three-smoking related diseases-lung cancer, ischaemic heart disease, and chronic obstructive lung disease. DESIGN: A prospective study of 21520 men aged 35-64 years when recruited in 1975-82 with detailed history of smoking and measurement of carboxyhaemoglobin. MAIN OUTCOME MEASURES: Notification of deaths (to 1993) classified by cause. RESULTS: Pipe and cigar smokers who had switched from cigarettes over 20 years before entry to the study smoked less tobacco than cigarette smokers (8.1 g/day v 20 g/day), but they had the same consumption as pipe and cigar smokers who had never smoked cigarettes (8.1 g) and had higher carboxyhaemoglobin saturations (1.2% v 1.0%, P < 0.001), indicating that they inhaled tobacco smoke to a greater extent. They had a 51% higher risk of dying of the three smoking related diseases than pipe or cigar smokers who had never smoked cigarettes (relative risk 1.51; 95% confidence interval 0.96 to 2.38), a 68% higher risk than lifelong non-smokers (1.68; 1.16 to 2.45), a 57% higher risk than former cigarette smokers who gave up smoking over 20 years before entry (1.57; 1.04 to 2.38), and a 46% lower risk than continuing cigarette smokers (0.54; 0.38 to 0.77). CONCLUSION: Cigarette smokers who have difficulty in giving up smoking altogether are better off changing to cigars or pipes than continuing to smoke cigarettes. Much of the effect is due to the reduction in the quantity of tobacco smoked, and some is due to inhaling less. Men who switch do not, however, achieve the lower risk of pipe and cigar smokers who have never smoked cigarettes. All pipe and cigar smokers have a greater risk of lung cancer than lifelong non-smokers or former smokers. PMID- 9224128 TI - Prospective cohort study of factors influencing the relative weights of the placenta and the newborn infant. AB - OBJECTIVES: To determine the demographic, environmental, and medical factors that influence the relative weights of the newborn infant and the placenta and compare this ratio with other factors known to predispose to adult ill health. DESIGN: Prospective cohort study. SETTING: The tertiary referral centre for perinatal care in Perth, Western Australia. SUBJECTS: 2507 pregnant women who delivered a single live infant at term. MAIN OUTCOME MEASURES: Placental weight, birth weight, and the ratio of placental weight to birth weight. RESULTS: By multiple regression analysis the placental weight to birthweight ratio was significantly and positively associated with gestational age, female sex, Asian parentage, increasing maternal body mass index, increased maternal weight at booking, lower socioeconomic status, maternal anaemia, and increasing number of cigarettes smoked daily. There were no consistent relations between the placental weight to birthweight ratio and measures of newborn size. CONCLUSIONS: The ratio of placental weight to birth weight is not an accurate marker of fetal growth. In its role as a predictor of adult disease the ratio may be acting as a surrogate for other factors which are already known to influence health and may act before or after birth. Determining the role that relative growth rates of the fetus and placenta have in predisposing to adult disease requires prospective study to account for the many confounding variables which complicate this hypothesis. PMID- 9224129 TI - Diabetes in institutionalised elderly people: a forgotten population? PMID- 9224130 TI - What future for continuity of care in general practice? PMID- 9224131 TI - Statistics notes. Units of analysis. PMID- 9224132 TI - Reliability of health information for the public on the World Wide Web: systematic survey of advice on managing fever in children at home. AB - OBJECTIVE: To assess the reliability of healthcare information on the world wide web and therefore how it may help lay people cope with common health problems. METHODS: Systematic search by means of two search engines, Yahoo and Excite, of parent oriented web pages relating to home management of feverish children. Reliability of information on the web sites was checked by comparison with published guidelines. MAIN OUTCOME MEASURES: Minimum temperature of child that should be considered as fever, optimal sites for measuring temperature, pharmacological and physical treatment of fever, conditions that may warrant a doctor's visit. RESULTS: 41 web pages were retrieved and considered. 28 web pages gave a temperature above which a child is feverish; 26 pages indicated the optimal site for taking temperature, most recommending rectal measurement; 31 of the 34 pages that mentioned drug treatment recommended paracetamol as an antipyretic; 38 pages recommended non-drug measures, most commonly tepid sponging, dressing lightly, and increasing fluid intake; and 36 pages gave some indication of when a doctor should be called. Only four web pages adhered closely to the main recommendations in the guidelines. The largest deviations were in sponging procedures and how to take a child's temperature, whereas there was a general agreement in the use of paracetamol. CONCLUSIONS: Only a few web sites provided complete and accurate information for this common and widely discussed condition. This suggests an urgent need to check public oriented healthcare information on the internet for accuracy, completeness, and consistency. PMID- 9224133 TI - Commentary: measuring quality and impact of the World Wide Web. PMID- 9224134 TI - Colorectal cancer. PMID- 9224135 TI - ABC of mental health. Anxiety. PMID- 9224136 TI - GP budget holding in New Zealand: lessons for Britain and elsewhere? PMID- 9224138 TI - Implications of managed care for health systems, clinicians, and patients. AB - The rhetoric and realities of managed care are easily confused. The rapid growth of managed care in the United States has had many implications for patients, doctors, employers, state and federal programmes, the health insurance industry, major medical institutions, medical research, and vulnerable patient populations. It has restricted patients' choice of doctors and limited access to specialists, reduced the professional autonomy and earnings of doctors, shifted power from the non-profit to the for-profit sectors and from hospitals and doctors to private corporations. It has also raised issues about the future structuring and financing of medical education and research and about practice ethics. However, managed care has also accorded greater prominence to the assessment of patient satisfaction, profiling and monitoring of doctors' work, the use of clinical guidelines and quality assurance procedures and indicated the potential to improve the integration and outcome of care. PMID- 9224137 TI - New Zealand's health reforms: a clash of cultures. PMID- 9224139 TI - Maternal and child health in China. AB - China has made great progress in improving the health of women and children over the past two generations. The success has been attributed to improved living standards, public health measures, and good access to health services. Although overall infant and maternal mortality rates are relatively low there are large differences in patterns of mortality between urban and rural areas. The Chinese have developed a hierarchical network of maternal and child health services, with each level taking a supervisory and teaching role for the level below it. Maternal and child health in China came to international attention in 1995 with the promulgation of the maternal and child health law. In China this was seen as a means of prioritising resources and improving the quality of services, but in the West it was widely described as a law on eugenics. PMID- 9224140 TI - Rationing health care. Allocating resources only to treating present dangers would ruin preventive health care and jeopardise future lives. PMID- 9224141 TI - Rationing health care. Cost effectiveness should not be the only criterion in deciding on treatment. PMID- 9224142 TI - Rationing health care. Access to treatment should be equal, regardless of age. PMID- 9224143 TI - Prevalence of gastrointestinal symptoms after bacterial gastroenteritis. Bowel symptoms vary over time. PMID- 9224144 TI - Prevalence of gastrointestinal symptoms after bacterial gastroenteritis. Patients with the irritable bowel syndrome may underreport historical symptoms. PMID- 9224145 TI - Prevalence of gastrointestinal symptoms after bacterial gastroenteritis. Psychological factors were not assessed. PMID- 9224146 TI - Prevalence of gastrointestinal symptoms after bacterial gastroenteritis. Study did not include a control group. PMID- 9224147 TI - Dual publication of surgical abstracts is acceptable. PMID- 9224148 TI - General practitioners prefer to work in cooperatives for out of hours work. PMID- 9224149 TI - Trial of prescribing strategies in managing sore throat. Penicillin had no effect in patients negative for group A beta haemolytic streptococci. PMID- 9224150 TI - Trial of prescribing strategies in managing sore throat. Failure to show antibiotic effectiveness was due to inclusion of cases of sore throat of viral origin. PMID- 9224151 TI - Compliance and concordance with treatment. Coming to an understanding with patients and prepositions. PMID- 9224152 TI - Compliance and concordance with treatment. Treating the patient as a decision maker is not always appropriate. PMID- 9224153 TI - Donations of orthopaedic equipment also cause problems. PMID- 9224154 TI - Overcoming racism in the NHS. An authoritative central body would help. PMID- 9224155 TI - Overcoming racism in the NHS. Positive aspects of multi-ethnic teams should be emphasised. PMID- 9224156 TI - Treatment for haemophilia by calendar in Hungary. PMID- 9224157 TI - Quoting intermediate analyses can only mislead. PMID- 9224158 TI - All doctors are problem doctors. Doctors must admit that they are human. PMID- 9224159 TI - All doctors are problem doctors. Network of "medical buddies" is needed. PMID- 9224160 TI - Court ordered caesarean sections are discouraging women from seeking obstetric care. PMID- 9224161 TI - Conclusions about why doctors change their practice were not supported by the data. PMID- 9224162 TI - Gun availability and violent death. PMID- 9224164 TI - Maternal considerations in formulating HIV-related breast-feeding recommendations. PMID- 9224163 TI - AIDS--an update on the global dynamics. PMID- 9224165 TI - Dealing with tobacco--the implications of a legislative settlement with the tobacco industry. PMID- 9224166 TI - Comment: Gunsmoke--changing public attitudes toward smoking and firearms. PMID- 9224167 TI - Comment: ethical dilemmas in worldwide polio eradication programs. PMID- 9224168 TI - Comment: evaluating the effectiveness of hospital care. PMID- 9224169 TI - Flawed gun policy research could endanger public safety. AB - A highly publicized recent study by Lott and Mustard concludes that laws easing restrictions on licenses for carrying concealed firearms in public substantially reduce violent crime. Several serious flaws in the study render the authors' conclusions insupportable. These flaws include misclassification of gun-carrying laws, endogeneity of predictor variables, omission of confounding variables, and failure to control for the cyclical nature of crime trends. Most of these problems should bias results toward overestimating the crime-reducing effects of laws making it easier to carry concealed firearms in public. Lott and Mustard's statistical models produce findings inconsistent with criminological theories and well-established facts about crime, and subsequent reanalysis of their data challenges their conclusions. Public health professionals should understand the methodological issues raised in this commentary, particularly when flawed research could influence the introduction of policies with potentially deleterious consequences. PMID- 9224170 TI - Ethical dilemmas in current planning for polio eradication. AB - Intensification of polio eradication efforts worldwide raises concerns about costs and benefits for poor countries. A major argument for global funding is the high benefit-cost ratio of eradication; however, financial benefits are greatest for rich countries. By contrast, the greatest costs are borne by poor countries; the Pan American Health Organization has estimated that host countries bore 80% of costs for polio eradication in the Americas. The 1988 World Health Assembly resolution setting up the Polio Eradication Initiative carried the proviso that programs should strengthen health infrastructures. Drastic cuts in donor funding for health make this commitment even more important. Two international evaluations have reported both positive and negative effects of polio and Expanded Programme on Immunization programs on the functioning and sustainability of primary health care. Negative effects were greatest in poor countries with many other diseases of public health importance. If poor countries are expected to divert funds from their own urgent priorities, donors should make solid commitments to long-term support for sustainable health development. PMID- 9224171 TI - Infant survival, HIV infection, and feeding alternatives in less-developed countries. AB - OBJECTIVES: This study examines, in the context of the human immunodeficiency virus (HIV) epidemic, the effects of optimal breast-feeding, complete avoidance of breast-feeding, and early cessation of breast-feeding. METHODS: The three categories of breast-feeding were weighed in terms of HIV transmission and infant mortality. Estimates of the frequency of adverse outcomes were obtained by simulation. RESULTS: Avoidance of all breast-feeding by the whole population always produces the worst outcome. The lowest frequency of adverse outcomes occurs if no HIV-seropositive women breast-feed and all seronegative women breast feed optimally, given infant mortality rates below 100 per 1000 and relative risks of dying set at 2.5 for non-breast-fed compared with optimally breast-fed infants. For known HIV-seropositive mothers, fewer adverse outcomes result from early cessation than from prolonged breast-feeding if the hazard of HIV transmission through breast-feeding after 3 months is 7% or more, even at high mortality rates, given relative risks of dying set at 1.5 for early cessation compared with optimal duration of breast-feeding. CONCLUSIONS: The risk of HIV transmission through breast-feeding at various ages needs to be more precisely quantified. The grave issues that may accompany a possible decline in breast feeding in the less developed world demand evaluation. PMID- 9224172 TI - Preventing unintended pregnancy: the cost-effectiveness of three methods of emergency contraception. AB - OBJECTIVES: This study examined the cost-effectiveness of emergency contraceptive pills, minipills, and the copper-T intrauterine device (IUD) as emergency contraception. METHODS: Cost savings were modeled for both (1) a single contraceptive treatment following unprotected intercourse and (2) emergency contraceptive pills provided in advance. RESULTS: In a managed care (public payer) setting, a single treatment of emergency contraception after unprotected intercourse saves $142 ($54) with emergency contraceptive pills and $119 ($29) with minipills. The copper-T IUD is not cost-effective as an emergency contraceptive alone, but savings quickly accrue as use continues. Advance provision of emergency contraceptive pills to women using barrier contraceptives, spermicides, withdrawal, or periodic abstinence saves from $263 to $498 ($99 to $205) annually. CONCLUSIONS: Emergency contraception is cost-effective whether provided when the emergency arises or in advance to be used as needed. Greater use of emergency contraception could reduce the considerable medical and social costs of unintended pregnancies. PMID- 9224173 TI - Mortality related to sexually transmitted diseases in US women, 1973 through 1992. AB - OBJECTIVES: This study estimated the trends in mortality related to sexually transmitted diseases (STDs) and their sequelae in US women from 1973 through 1992. METHODS: The total number of deaths was obtained from US national mortality data and from AIDS surveillance data, and current literature was reviewed to estimate proportions of diseases attributable to sexual transmission. RESULTS: From 1973 through 1984, total STD-related deaths decreased 24%. However, from 1985 through 1992, STD-related deaths increased by 31%, primarily because of increasing numbers of deaths from sexually transmitted human immunodeficiency virus (HIV) infection. The most important changes during the 20-year period were the emergence of and continued increase in the number of deaths related to hetero sexually transmitted HIV. CONCLUSIONS: The leading causes of STD-related mortality in women, viral STDs and their sequelae, are generally not recognized as being sexually transmitted. Increases in STD-related mortality are primarily due to sexually transmitted HIV, which will soon surpass cervical cancer as the leading cause. PMID- 9224174 TI - Maternal employment and breast-feeding: findings from the 1988 National Maternal and Infant Health Survey. AB - OBJECTIVES: This analysis uses nationally representative data from the 1988 National Maternal and Infant Health Survey to explore the factors, including employment, associated with breast-feeding initiation and duration. METHODS: Multiple logistic regression was used to model the determinants of breast-feeding initiation among 9087 US women. Multiple linear regression was used to model the duration of breast-feeding among women who breast-fed. RESULTS: Fifty-three percent of mothers initiated breast-feeding in 1988, and the decision to breast feed was not associated with maternal employment. However, among breast-feeders, returning to work within a year of delivery was associated with a shorter duration of breast-feeding when other factors were controlled. Among employed mothers, the duration of maternity leave was positively associated with the duration of breast-feeding. CONCLUSIONS: The low rates of breast-feeding initiation in the United States are not attributable to maternal participation in the labor force. However, returning to work is associated with earlier weaning among women who breast-feed. PMID- 9224175 TI - Some correlates of self-rated health for Australian women. AB - OBJECTIVES: This study aimed to identify some of the correlates of self-rated health for young to middle-aged Australian women. METHODS: Regression analyses were based on a 4-year longitudinal study using a random sample of Sydney women 20 to 59 years of age at baseline. Participants were interviewed in 1986/87 and 1990. RESULTS: Cross-sectional relationships between self-assessed health and other health measures varied significantly by age, although physical health was a common correlate. Sixty-three percent of participants reported a similar rating of health over the 4-year period between the surveys. Changes in self-assessed health were sensitive to chronic disease. Also, participants' self-ratings of health were related to their subsequent chronic disease status. CONCLUSIONS: Self rated health reflects a complex process of internalized calculations that encompass both lived experience and knowledge of disease causes and consequences. Women seem to take into consideration a broad range of factors, including lifestyle, vitality, mental attitude, and age, and, if they have a health condition, the chronicity of their disease, duration since diagnosis, and treatment. PMID- 9224176 TI - Frequent attendance at religious services and mortality over 28 years. AB - OBJECTIVES: This study analyzed the long-term association between religious attendance and mortality to determine whether the association is explained by improvements in health practices and social connections for frequent attenders. METHODS: The association between frequent attendance and mortality over 28 years for 5286 Alameda Country Study respondents was examined. Logistic regression models analyzed associations between attendance and subsequent improvements in health practices and social connections. RESULTS: Frequent attenders had lower mortality rates than infrequent attenders (relative hazard [RH] = 0.64;95% confidence interval [CI] = 0.53,0.77). Results were stronger for females. Health adjustments had little impact, but adjustments for social connections and health practices reduced the relationship (RH = 0.77; 95% CI = 0.64, 0.93). During follow-up, frequent attenders were more likely to stop smoking, increase exercising, increase social contacts, and stay married. CONCLUSIONS: Lower mortality rates for frequent religious attenders are partly explained by improved health practices, increased social contacts, and more stable marriages occurring in conjunction with attendance. The mechanisms by which these changes occur have broad intervention implications. PMID- 9224178 TI - The campaign to raise the tobacco tax in Massachusetts. AB - OBJECTIVES: Question 1 raised the Massachusetts state tobacco tax to fund tobacco education programs. This paper examines the process of qualifying and passing Question 1. METHODS: Information was gathered from internal memoranda, meeting minutes, newspaper articles, internal documents, letters, newsletters, news and press releases, and personal interviews. Data about campaign contributions were obtained from the Massachusetts Office of Campaign and Political Finance. RESULTS: Three factors help explain why Question 1 passed: (1) the policy environment was favorable because of the social unacceptability of smoking; (2) the activists assembled a large coalition of supporters; and (3) the activists countered industry claims that the new tax would hurt small business and lower income smokers and would be wasted by the legislature. The ballot initiative passed despite the industry's $7 million campaign to defeat it. CONCLUSIONS: The apparent influence of the tobacco industry on the legislature was the driving force behind the decision of public health activists to qualify Question 1. Moving policy-making out of the legislature into the public arena widened the scope of conflict and enabled public health activists to win. PMID- 9224177 TI - Routine prenatal screening for congenital heart disease: what can be expected? A decision-analytic approach. AB - OBJECTIVES: This study assessed the potential impact of fetal ultrasound screening on the number of newborns affected by cardiac anomalies. METHODS: A decision model was developed that included the prevalence and history of congenital heart disease, characteristics of ultrasound, risk of abortion, and attitude toward pregnancy termination. Probabilities were obtained with a literature survey; sensitivity analysis showed their influence on expected outcomes. RESULTS: Presently, screening programs may prevent the birth of approximately 1300 severely affected newborns per million second-trimester pregnancies. However, over 2000 terminations of pregnancy would be required, 750 of which would have ended in intrauterine death or spontaneous abortion. Further, 9900 false-positive screening results would occur, requiring referral. Only the sensitivity of routine screening and attitude toward termination of pregnancy appeared to influence the yield substantially. CONCLUSIONS: The impact of routine screening for congenital heart disease appeared relatively small. Further data may be required to fully assess the utility of prenatal screening. PMID- 9224179 TI - The association between the purchase of a handgun and homicide or suicide. AB - OBJECTIVES: The purpose of this study was to determine whether purchase of a handgun from a licensed dealer is associated with the risk of homicide or suicide and whether any association varies in relation to time since purchase. METHODS: A case-control study was done among the members of a large health maintenance organization. Case subjects were the 353 suicide victims and 117 homicide victims among the members from 1980 through 1992. Five control subjects were matched to each case subject on age, sex, and zip code of residence. Handgun purchase information was obtained from the Department of Licensing. RESULTS: The adjusted relative risk of suicide was 1.9 (95% confidence interval [CI] = 1.4, 2.5) for persons with a history of family handgun purchase from a registered dealer. The adjusted relative risk for homicide, given a history of family handgun purchase, was 2.2 (95% CI = 1.3, 3.7). For both suicide and homicide, the elevated relative risks persisted for more than 5 years after the purchase. CONCLUSIONS: Legal purchase of a handgun appears to be associated with a long-lasting increased risk of violent death. PMID- 9224180 TI - The effectiveness of a school-based curriculum for reducing violence among urban sixth-grade students. AB - OBJECTIVES: In this study, we examine the impact of a school-based curriculum designed to reduce violence among urban sixth-grade students. METHODS: This study used a staggered implementation design in which sixth-graders in six middle schools were taught an 18-session violence-prevention curriculum during either the fall or spring semester. Outcome measures were completed at the beginning, middle, and end of the school year. RESULTS: For boys, participation in the program during the fall resulted in significant posintervention differences in the self-reported frequency of violence and several other problem behaviors. Most of these differences were maintained at the end of the school year. Girls, in contrast, did not appear to benefit from the program. CONCLUSION: These results support the use of a school-based curriculum for reducing violence among sixth grade boys. They also underscore the importance of early intervention and the necessity of examining gender effects in evaluating such programs. PMID- 9224181 TI - Profiles of violent youth: substance use and other concurrent problems. AB - OBJECTIVES: This study examined the prevalence of various violent behaviors among high school-age adolescents, the co-occurrence of teenage violence with other public health problems, and gender differences in violence. METHODS: Longitudinal data for more than 4500 high school seniors and dropouts from California and Oregon were used to develop weighted estimates of the prevalence of violent behavior and its co-occurrence with other emotional and behavioral problems. RESULTS: More than half the sample had engaged in violence during the last year, and one in four had committed predatory violence. Boys were more likely than girls to engage in most types of violence, but both were equally prone to violence within the family. Violent youth were more likely than their peers to have poor mental health, use drugs, drop out of school, and be delinquent. Violent boys were more likely than violent girls to commit nonviolent felonies and sell drugs, but less likely to have poor mental health or become a parent. Prevalence estimates for violence co-occurring with three or more other problems ranged from 4% to 21%. CONCLUSIONS: Teenage violence typically coexists with additional emotional and behavioral problems. Programs must consider the broader public health context in which violence occurs. PMID- 9224182 TI - Milk, dietary calcium, and bone fractures in women: a 12-year prospective study. AB - OBJECTIVES: This study examined whether higher intakes of milk and other calcium rich foods during adult years can reduce the risk of osteoporotic fractures. METHODS: This was a 12-year prospective study among 77761 women, aged 34 through 59 years in 1980, who had never used calcium supplements. Dietary intake was assessed with a food-frequency questionnaire in 1980, 1984, and 1986. Fractures of the proximal femur (n = 133) and distal radius (n = 1046) from low or moderate trauma were self-reported on biennial questionnaires. RESULTS: We found no evidence that higher intakes of milk or calcium from food sources reduce fracture incidence. Women who drank two or more glasses of milk per day had relative risks of 1.45 for hip fracture (95% confidence interval [CI] = 0.87, 2.43) and 1.05 for forearm fracture (95% CI = 0.88, 1.25) when compared with women consuming one glass or less per week. Likewise, higher intakes of total dietary calcium or calcium from dairy foods were not associated with decreased risk of hip or forearm fracture. CONCLUSIONS: These data do not support the hypothesis that higher consumption of milk or other food sources of calcium by adult women protects against hip or forearm fractures. PMID- 9224183 TI - Determining injury at work on the California death certificate. AB - OBJECTIVES: This study examined decisions of California Country Coroner's offices in determining injury at work and identified factors influencing this decision. METHODS: Surveys were sent to California County Coroner's offices (response rate = 93%). The survey included 23 vignettes that required the respondent to determine whether the fatality involved an injury at work. The Rasch method was used to determine internal consistency in endorsing vignettes and to determine overall endorsability of vignettes based on underlying factors. RESULTS: Respondents showed internal consistency but much disagreement in their endorsement of vignettes. Decedents who were performing paid work or were on their work site during working hours were almost unanimously endorsed as having incurred an injury at work. Non-payment, travel/transportation, suicide, and nontraditional work sites and work hours led to disagreement and uncertainty among respondents. CONCLUSIONS: Coroners have different methods of determining injury at work on the death certificate, and available guidelines do not define many of the ambiguous situations encountered by coroners. PMID- 9224184 TI - The effect of high altitude and other risk factors on birthweight: independent or interactive effects? AB - OBJECTIVES: This study examined whether the decline in birth-weight with increasing altitude is due to an independent effect of altitude or an exacerbation of other risk factors. METHODS: Maternal, paternal, and infant characteristics were obtained from 3836 Colorado birth certificates from 1989 through 1991. Average altitude of residence for each county was determined. RESULTS: None of the characteristics related to birthweight (gestational age, maternal weight gain, parity, smoking, prenatal care visits, hypertension, previous small-for-gestational-age infant, female newborn) interacted with the effect of altitude. Birthweight declined an average of 102 g per 3300 ft (1000 m) elevation when the other characteristics were taken into account, increasing the percentage of low birthweight by 54% from the lowest to the highest elevations in Colorado. CONCLUSIONS: High altitude acts independently from other factors to reduce birthweight and accounts for Colorado's high rate of low birthweight. PMID- 9224185 TI - Reduced probability of HIV infection among crack cocaine--using injection drug users. AB - OBJECTIVES: This study examined in greater detail the authors' previously reported finding that crack use among injection drug users is associated with lower levels of infection with the human immunodeficiency virus (HIV). METHODS: Self-reported data and blood tests for HIV antibodies from 4840 out-of-treatment injection drug users were used to examine relationships among crack use, HIV risk behavior, and HIV infection. RESULTS: Crack use was significantly associated with higher levels of many sexual risk and needle use behaviors and was consistently associated, independently of all behavioral variables examined, with lower rates of HIV infection. CONCLUSIONS: Crack use among injection drug users appears to be associated with lower risk for HIV infection independently of other behavioral variables. PMID- 9224186 TI - Low prevalences of HIV infection and sexually transmitted disease among female commercial sex workers in Mexico City. AB - OBJECTIVES: This study tried to determine human immunodeficiency virus (HIV)/sexually transmitted disease (STD) prevalences among female commercial sex workers in Mexico City. METHODS: A sampling frame was constructed that included bars, massage parlors, and street corners. RESULTS: Prevalences for Treponema pallidum, herpes simplex virus type 2, HIV, Neisseria gonorrhoeae, and Chlamydia trachomatis were 6.4%, 65%, 0.6%, 3.7%, and 11.1%, respectively. A significant association was found between higher STD frequencies and working at street sites. CONCLUSIONS: Most STD frequencies were lower in comparison with rates found for female sex workers in other countries. However, preventive programs against STD/ HIV are needed in this population. PMID- 9224187 TI - The effects of an abusive primary partner on the condom use and sexual negotiation practices of African-American women. AB - OBJECTIVES: This study examined the consequences of having a physically abusive primary partner on the condom use and sexual negotiation practices of young African-American women. METHODS: Interviews were conducted with 165 sexually active African-American women aged 18 through 29 in San Francisco, Calif. RESULTS: Women in abusive relationships were less likely than others to use condoms and were more likely to experience verbal abuse, emotional abuse, or threats of physical abuse when they discussed condoms. They were more fearful of asking their partners to use condoms, worried more about acquiring the human immunodeficiency virus (HIV), and felt more isolated than did women not in abusive relationships. CONCLUSIONS: HIV prevention programs for women should address domestic violence prevention strategies. PMID- 9224188 TI - The decentralization of syphilis screening for improved care in Jamaican public clinics. Collaborative Working Group on Decentralized Syphilis Screening. AB - OBJECTIVES: This study examined the decentralization of syphilis screening for improved care in Jamaican public clinics. METHODS: One of every five serum samples tested at the six peripheral sites was frozen and retested at the central laboratory in Kingston. Patient files and laboratory logbooks were compared over a 3-month period. RESULTS: Between May 1993 and December 1994, 15.5% of 32913 patients with sexually transmitted diseases and 8.3% of 8914 women seeking prenatal care were found syphilis seroreactive. Of 2001 samples evaluated, 1933 (96.6%) had been correctly reported at the peripheral sites. Of 129 syphilis seroreactors detected at the peripheral sites, 88 (68%) were treated the same day and 21 (16%) more within 3 days after testing. CONCLUSIONS: Syphilis seroreactors were accurately detected and quickly treated at the peripheral sites. If these efforts can be sustained, Jamaican syphilis rates should decrease. PMID- 9224189 TI - Prenatal health behaviors and psychosocial risk factors in pregnant women of Mexican origin: the role of acculturation. AB - OBJECTIVES: This study examined the association between acculturation of Mexican origin women and factors in low birthweight and preterm delivery. METHODS: Interviews were conducted with 911 Mexican-origin respondents in Los Angeles prenatal care clinics. Infant outcome data were retrieved from delivery records. RESULTS: Mexican-American women had generally more undesirable prenatal behaviors and risk factors than Mexican-immigrant women. Although higher acculturation was significantly associated with behavioral risk factors, there were no direct effects of acculturation on infant gestational age or birthweight. CONCLUSIONS: Future research needs to measure multiple factors to assess their effects on culture-specific protective factors. PMID- 9224190 TI - The relationship of patient reading ability to self-reported health and use of health services. AB - OBJECTIVES: This study examined the relationship of functional health literacy to self-reported health and use of health services. METHODS: Patients presenting to two large, urban public hospitals in Atlanta, Ga, and Torrance, Calif, were administered a health literacy test about their overall health and use of health care services during the 3 months preceding their visit. RESULTS: Patients with inadequate functional health literacy were more likely than patients with adequate literacy to report their health as poor. Number of years of school completed was less strongly associated with self-reported health. Literacy was not related to regular source of care or physician visits, but patients in Atlanta with inadequate literacy were more likely than patients with adequate literacy to report a hospitalization in the previous year. CONCLUSIONS: Low literacy is strongly associated with self-reported poor health and is more closely associated with self-reported health than number of years of school completed. PMID- 9224191 TI - The effectiveness of a media-led intervention to reduce smoking among Vietnamese American men. AB - OBJECTIVES: This study evaluated an anti-tobacco campaign targeting Vietnamese men in San Francisco, Calif. METHODS: The intervention included Vietnamese language media, health education materials, and activities targeting physicians, youth, and businesses. Evaluation involved pretest and posttest cross-sectional telephone surveys and multiple logistic regression analyses designed to identify variables associated with smoking and quitting. RESULTS: At posttest, the odds of being a smoker were significantly lower (odds ratio [OR] = 0.82, 95% confidence interval [CI] = 0.68, 0.99), and the odds of being a quitter were significantly higher (OR = 1.65, 95% CI = 1.27, 2.15), in San Francisco than in a comparison community. CONCLUSIONS: Despite modest success, further efforts are needed to reduce smoking among Vietnamese-American men. PMID- 9224192 TI - Employee and public responses to simulated violations of no-smoking regulations in Spain. AB - OBJECTIVES: This study evaluated compliance with regulations prohibiting smoking in public places by using a new method of smoking simulation. METHODS: Sites in Sabadell, Spain, were visited by observers who lit a cigarette, simulating the act of smoking, and noted the placement and content of signs indicating smoking restrictions. RESULTS: A warning was given in only 17% of sites. A significant association was observed between the presence of signs banning smoking and the elicitation of a warning from people present. CONCLUSIONS: Simulating the violation of smoking restrictions may be used as an efficient method of assessing enforcement of the prohibition of public smoking. PMID- 9224194 TI - Trends in fatal occupational injuries and industrial restructuring in North Carolina in the 1980s. AB - OBJECTIVES: This study examined the relationship between changes in employment in North Carolina in the 1980s and fatal occupational injury rates. METHODS: Unintentional fatal occupational injuries (n = 1989) in North Carolina between 1978 and 1991 were identified via the medical examiner's system. RESULTS: Overall fatal injury rates declined during the 1980s, but rates increased 9.6% per year among manufacturing industries that declined in employment size; rates fell among service sector and manufacturing industries that grew. CONCLUSIONS: Increasing occupational fatal injury rates accompanied the decline in workforce in North Carolina's traditional, labor-intensive manufacturing industries during the 1980s, while service sector and expanding manufacturing industries have experienced declining fatal injury rates. PMID- 9224195 TI - Increasing car seat use for toddlers from inner-city families. AB - OBJECTIVES: The purpose of this project was to increase toddler car seat use in low-income minority families. METHODS: Families from Newark, NJ, were divided into two study groups. Both groups were given car seats; one group also received education regarding car restraint use. Observations were made of car seat use before car seat distribution, immediately after distribution, 4 to 5 months later, and 1 year later. RESULTS: Car seat use increased markedly immediately after distribution and remained high 1 year later, regardless of education. CONCLUSIONS: These results indicate that distributing car seats results in long term use among a currently low-use population. PMID- 9224193 TI - Weapon carrying among black adolescents: a social network perspective. AB - OBJECTIVES: This report describes the salience of social networks to the phenomena of adolescent weapon carrying. METHODS: A random-walk network sampling design was used to survey 113 adolescents about topics, including weapon carrying. RESULTS: In a probability sample of 12- to 15-year-olds, 20.9% reported ever carrying a weapon. Carriers were eight times as likely as noncarriers to report weapon carrying by an older associate, and 19 times as likely to report weapon carrying by a peer. A significant dose-response effect was present. CONCLUSIONS: This evidence supports the interpretation that modeling of weapon carrying by personal network members is important for its initiation and maintenance in adolescence. PMID- 9224196 TI - Assessing children's ultraviolet radiation exposure: the use of parental recall via telephone interviews. AB - OBJECTIVES: This study evaluated the validity of a parental report measure of children's solar protection behaviors. METHODS: Fifty-eight children had skin color assessed twice with a colorimeter. Between measurement sessions, parents were interviewed by telephone to assess children's indoor-outdoor status and solar protection across 40 hourly intervals. RESULTS: Parental report of child's indoor-outdoor status was significantly correlated with the colorimeter values, whereas the use of sunscreen and protective clothing was not. CONCLUSIONS: This measure was feasible for assessing ultraviolet exposure in young children. The component that assessed the number of intervals spent outdoors evidenced predictive validity. PMID- 9224197 TI - Injuries to bicyclists in Wuhan, People's Republic of China. AB - OBJECTIVES: This study examined the morbidity and mortality from bicycling injuries in Wuhan, China. METHODS: Police department data for the year 1993 complemented by data from emergency room interviews were analyzed. RESULTS: The death rate from bicycling injuries was estimated as 2.2 per 100000 population, more than seven times the rate for the United States. At least 79% of the fatalities and 17% of the emergency room cases sustained head injuries, the majority (71%) of which resulted from contact of the head with the concrete or asphalt road. None of the patients was wearing a helmet at the time of injury, and helmet use among the general bicyclist population was nonexistent. CONCLUSIONS: Bicycle-related head injury is an important public health issue in China. The effectiveness of safety helmets in developing countries needs to be evaluated. PMID- 9224199 TI - Condom-carrying behavior among college students. PMID- 9224198 TI - Medical necessity and defined coverage benefits in the Oregon Health Plan. AB - The policy debate in Oregon has primarily focused on the Prioritized List of Services. However, little information is available on how defined coverage benefits and managed care affect the role of medical necessity in determining care for Medicaid patients. This issue is important because medical necessity determinations are currently used by many states to limit extraneous health care costs but require resource-intensive oversight, are open to wide variance, and frequently prompt litigation challenging interpretations of what is necessary and what is not. The qualitative study described here addressed whether medical necessity remains a salient and useful concept in the Oregon Health Plan. Our results indicate that defined coverage benefits, as described by the funded portion of the Prioritized List of Services, supplant medical necessity determinations for coverage, while managed care incentives limit the need for medical necessity determinations at the provider level. Clinical choices are, for the most part, guided by providers' judgment within the financial constraints of capitation and by targeted use management techniques. The combination of capitated care and Oregon's defined coverage benefits package has marginalized the use of medical necessity, albeit with consequences for state oversight of Medicaid services. PMID- 9224200 TI - A call for consistency in defining breast-feeding. PMID- 9224201 TI - Improving Americans' diet. PMID- 9224202 TI - Posttranscriptional regulation of lung elastin production. PMID- 9224203 TI - The role of nuclear factor-kappa B in cytokine gene regulation. AB - Transcription factors are DNA-binding proteins that regulate gene expression. Nuclear factor-kappa B (NF-kappa B) is a critical transcription factor for maximal expression of many cytokines that are involved in the pathogenesis of inflammatory diseases, such as adult respiratory distress syndrome (ARDS) and sepsis syndrome. Activation and regulation of NF-kappa B are tightly controlled by a group of inhibitory proteins (I kappa B) that sequester NF-kappa B in the cytoplasm of immune/inflammatory effector cells. NF-kappa B activation involves signaled phosphorylation, ubiquitination, and proteolysis of I kappa B. Liberated NF-kappa B migrates to the nucleus, where it binds to specific promoter sites and activates gene transcription. The activation of NF-kappa B initiates both extracellular and intracellular regulatory events that result in autoregulation of the inflammatory cascade through modulation of NF-kappa B activation. Recently, activation of NF-kappa B has been linked to ARDS and has been shown to be a critical proximal step in the initiation of neutrophilic inflammation in animal models. Activation of NF-kappa B can be inhibited in vivo by treatment with antioxidants, corticosteroids, and the induction of endotoxin tolerance. Identification of more specific and efficacious inhibitors of NF-kappa B activation might prove beneficial for the treatment of cytokine-mediated inflammatory diseases. PMID- 9224204 TI - Stabilization of elastin mRNA by TGF-beta: initial characterization of signaling pathway. AB - The cytokine transforming growth factor-beta (TGF-beta) has multiple effects on a wide variety of cell types. These effects include modulation of growth and regulation of gene transcription. In a few instances, TGF-beta has also been shown to regulate gene expression posttranscriptionally by altering message stability, but the pathway by which this activity is executed remains largely unknown. In the present work, we demonstrate that TGF-beta 1 has no effect on transcription of the elastin gene in cultured human fetal lung fibroblasts, but does stabilize elastin messenger RNA (mRNA), leading to a dramatic increase in the steady-state level of elastin mRNA. A corresponding increase in production of tropoelastin accompanies the increase in elastin mRNA. Through the use of specific inhibitors, we demonstrate that phosphatidylcholine (PC)-specific phospholipase C (PLC) and protein kinase C (PKC) are involved in mediating the elastin message stabilization. In contrast, G proteins and extracellularly regulated kinases do not appear to be involved. These results suggest that although the TGF-beta signaling pathway leading to message stabilization shares components with that modulating transcription, the message-stabilization pathway also contains diverse other elements. PMID- 9224206 TI - Exogenous and endogenous transforming growth factors-beta influence elastin gene expression in cultured lung fibroblasts. AB - Elastin, an important structural protein of the extracellular matrix, confers elastic properties on the pulmonary alveolar interstitium. In the alveolar wall, elastin is primarily produced postnatally by fibroblasts. The mechanisms that regulate lung fibroblast (LF) elastin gene expression have not been completely defined, although both transcriptional and posttranscriptional mechanisms appear to be involved. Transforming growth factors-beta (TGF-beta s) have been shown to increase elastin production by cultured neonatal rat LF. Analyses of elastin gene transcription and mRNA stability indicate that exogenous TGF-beta 1 increases the half-life of tropoelastin mRNA by 1.5-fold and does not alter elastin gene transcription. Interference with the functions of endogenous TGF-beta 1 in cultured LF, through the addition of neutralizing antibodies or antisense oligodeoxynucleotides, decreases tropoelastin and tropoelastin mRNA production by these cells. The content of total (latent plus active) TGF-beta s was approximately 4.5-fold greater in lungs obtained from rats on postnatal day 8 than in lungs obtained from adults. These findings indicate that endogenous TGF beta s, in cultured LF, regulate elastin gene expression, most likely by a posttranscriptional mechanism. Since others have shown that elastin mRNA appears to have a longer half-life in neonatal than in adult rat lungs, we hypothesize that the higher content of TGF-beta s could contribute to the greater elastin mRNA stability in neonatal lungs. PMID- 9224205 TI - IL-13 mRNA and immunoreactivity in allergen-induced rhinitis: comparison with IL 4 expression and modulation by topical glucocorticoid therapy. AB - The allergen-induced late nasal response (LNR) is associated with high expression of interleukin-4 (IL-4) and IL-5 messenger RNA (mRNA) in the nasal mucosa, suggesting a role for Th2-type cytokines in the development of the LNR. Moreover, topical corticosteroid-mediated inhibition of the LNR is accompanied by inhibition of IL-4, but not IL-5, mRNA expression, IL-13 shares a number of functions with IL-4, including IgE switching and vascular cell adhesion molecule 1 (VCAM-1) upregulation. We investigated the expression of IL-13 mRNA and immunoreactivity in nasal biopsies from 10 normal subjects and 20 subjects with allergic rhinitis. IL-4 mRNA expression was examined in the same subjects. The allergic rhinitis patients were randomized to receive a 6-wk treatment with either topical fluticasone propionate (n = 10) or placebo (n = 10) nasal spray twice daily. A nasal biopsy was taken before treatment and 24 h after local nasal allergen provocation with a grass-pollen extract. Before treatment, there was no significant difference between the allergic rhinitis patients and controls in the expression of IL-13 mRNA and immunoreactivity. After allergen provocation, we observed a significant increase in IL-13 mRNA-positive and immunoreactive cells at 24 h only in subjects given placebo (P < 0.001). Inhibition of the LNR after corticosteroid treatment was associated with a marked decrease in allergen induced IL-13 mRNA-positive (P < 0.001) and immunoreactive cells (P < 0.001). In subjects given placebo, 76.9 +/- 5.5% of IL-13 mRNA-positive cells observed after allergen were CD3+, whereas 11.2 +/- 2.7% coexpressed immunoreactivity for mast cell tryptase. In these subjects, increases in cells expressing IL-13 mRNA were greater than for IL-4 mRNA (P = 0.001), and double in situ hybridization studies revealed that 100% of the IL-4 mRNA-positive cells coexpressed IL-13 mRNA, whereas 66.6 +/- 10.5% of IL-13 mRNA-positive cells coexpressed IL-4 transcripts after allergen challenge. The results of this study suggest that IL-13 expression is a prominent feature of the LNR, and that inhibition of the LNR following steroid therapy may be partly attributable to inhibition of IL-13 expression. PMID- 9224207 TI - Inducible nitric oxide synthase mRNA and immunoreactivity in the lungs of rats eight hours after antigen challenge. AB - We have previously shown that inducible nitric oxide (iNO)-synthase immunoreactivity is expressed in bronchial epithelium and increased in asthma which suggests a possible role for NO in airway hyperresponsiveness. We tested the hypothesis that exposure of a sensitized animal to antigen could account for the increased expression of iNO-synthase in the airways. We examined the expression of iNO-synthase mRNA and immunoreactivity in the lungs of ovalbumin (OA) sensitized Brown Norway (BN) rats 8 h after antigen challenge by in situ hybridization and immunocytochemistry. Sensitized and unchallenged or bovine serum albumin (BSA) challenged rats, or unsensitized and OA challenged rats served as controls. With the use of an iNO-synthase probe we found a higher expression of iNO-synthase mRNA in BN rat airways after antigen challenge with OA but not after antigen challenge with BSA or in other controls. Most of the expression was in the epithelium of the airways with few cells positive in the subepithelial inflammatory infiltrate or in lung lavage. Very strong iNO-synthase immunoreactivity was observed in the airway epithelium of sensitized and OA challenged rats. No significant immunoreactivity was observed in the inflammatory infiltrate of the airways or in lung parenchyma. In conclusion, iNO-synthase increases in the airways of sensitized rats after exposure to antigen, the major source being from airway epithelial cells. NO may have a role in the development of the late airway response and bronchial hyperresponsiveness. PMID- 9224208 TI - Conductive airway surfactant: surface-tension function, biochemical composition, and possible alveolar origin. AB - Alveolar surfactant is well known for its ability to reduce minimal surface tension at the alveolar air-liquid interface to values below 5 mN/m. In addition, it has been suggested that an analogous conductive airway surfactant is also present in the airways. To elucidate the composition, possible origin, and surface activity of conductive airway phospholipids (PL), we compared in adult porcine lungs the PL classes and phosphatidylcholine (PC) molecular species of nonpurified tracheal aspirate samples with those of bronchoalveolar lavage fluid (BAL), tracheobronchial epithelium, and lung parenchyma. We also analyzed PL and PC composition, protein content, and surface activity of surfactant isolated from tracheal aspirates (SurfTrachAsp), BAL (SurfBAL), and the 27,000 x g pellet of BAL (SurfP27000) by density-gradient centrifugation. Although PL composition revealed contributions of the airways to tracheal aspirates, the composition of PC molecular species of tracheal aspirates was similar to that of BAL and lung parenchyma, but differed considerably from that of airway epithelium. SurfTrachAsp had the same PL and PC composition as SurfBAL and SurfP27000, indicating that this fraction of tracheal aspirates may have originated from the alveoli. Nevertheless, minimal and maximal surface tensions were higher in SurfTrachAsp than in SurfBAL and SurfP27000. Analysis of surfactant proteins A, B, and C (SP-A, SP-B, and SP-C) revealed that SP-A was decreased and SP-B and SP C were absent, whereas total protein was increased in SurfTrachAsp. We conclude that as compared with alveolar surfactant, PL of SurfTrachAsp show the same composition, but that surface-tension function is impaired and the concentration of surfactant proteins is decreased in SurfTrachAsp. PMID- 9224209 TI - Interaction with type II estrogen binding sites and antiproliferative activity of tamoxifen and quercetin in human non-small-cell lung cancer. AB - The antiestrogen tamoxifen is thought to antagonize the effects of estrogens by competing with them for estrogen receptor (ER) binding. However, tarnoxifen can also reverse multidrug resistance, synergize with cisplatin cytotoxicity, and inhibit growth in ER-negative lung cancer cells. In addition to ERs, rat and human target tissues contain a second binding macromolecule termed the type II estrogen binding site (type II EBS). It has been shown that tamoxifen and flavonoids, a widely distributed class of natural substances with a variety of biologic actions, bind to type II EBS and inhibit the growth of several tumor cell types. At present, conflicting data about ERs and an absence of data about type II EBSs exist for lung tumors. We have tested non-small-cell lung carcinoma cell lines and primary tumor cells for the presence of ERs and type II EBSs and have evaluated the effects of tamoxifen and quercetin (pentahydroxyflavone) on the growth of these cells. Using a whole-cell assay and nuclear and cytosolic radiobinding experiments with [3H]estradiol as tracer, we have found that SK-LU1, SW900, ChaGo-K-1, H441, H661, and A549 cells, as well as primary tumors, bind estrogen specifically. This binding results mainly from the presence of a large number of type II EBSs, whereas ERs are absent or present at low concentrations. Type II EBSs bound tamoxifen and quercetin with similar affinity. Cell counts and a thymidine incorporation assay showed that both compounds inhibit cell growth in a concentration-dependent manner at concentrations ranging from 10 nM to 1 microM. Neither ipriflavone, an isoflavone, nor rutin, the 3-rhamnosylglucoside of quercetin, bound type II EBSs or inhibited cell growth. These findings suggest that tamoxifen and quercetin could regulate lung cancer cell growth through a binding interaction with type II EBSs. This mechanism could also be active in vivo, in that we have observed that nuclear and cytosolic type II EBSs were present in all primary lung cancers tested (n = 12), and that tamoxifen and quercetin were effective in inhibiting in vitro bromodeoxyuridine (BrdU) incorporation and proliferation-cell nuclear antigen expression by neoplastic cells in these cancers. PMID- 9224210 TI - Polarized secretion of fibrinogen by lung epithelial cells. AB - The lung epithelium has recently been identified as a novel site of fibrinogen (FBG) biosynthesis. A coordinated upregulation of A alpha, B beta, and gamma chain FBG gene transcription occurs upon stimulation of A549 lung epithelial cells with dexamethasone (DEX) and the proinflammatory mediator interleukin-6 (IL 6). Subsequently, the cells synthesize and secrete fully assembled FBG. This study addresses the polarity of such FBG secretion by A549 cells cultured on polycarbonate membrane filters. After induction with IL-6 and DEX, cells were metabolically labeled, and FBG was immunopurified from the apical and basolateral chambers. Analysis by gel electrophoresis revealed that A549 cells secreted newly synthesized FBG in a polarized manner, with the majority (80%) of FBG secreted basolaterally. Consistent with this observation, immunoelectron microscopy using Protein A-gold labeling showed FBG within secretory vesicles in close proximity to the basolateral aspect of the A549 cell membrane. Polarized secretion was microtubule-dependent since depolymerization using colchicine significantly reduced the basolateral component of secretion, causing intracellular retention of FBG. These data provide evidence that FBG is secreted by lung alveolar epithelial cells vectorially toward the basement membrane, which may reflect in vivo processes associated with local injury, inflammation, and repair mechanisms. PMID- 9224211 TI - Expression of vascular endothelial growth factor by human eosinophils: upregulation by granulocyte macrophage colony-stimulating factor and interleukin 5. AB - Vascular endothelial growth factor (VEGF) is a pleiotropic polypeptide that mediates endothelial-cell-specific responses such as induction of proliferation and vascular leakage. We examined the expression of VEGF messenger RNA (mRNA) and protein by human eosinophils in response to granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-5 (IL-5). Immunoreactive VEGF protein was detected in freshly isolated eosinophils by immunocytochemistry. Eosinophils spontaneously released VEGF protein in culture medium, and this release was upregulated by GM-CSF or IL-5. Freshly isolated eosinophils constitutively expressed VEGF mRNA. Although incubation of eosinophils in culture medium reduced steady-state VEGF mRNA levels, eosinophil VEGF mRNA levels were enhanced by GM CSF and IL-5, and this enhancement was blocked by the transcription inhibitor actinomycin D. Analysis of alternatively spliced mRNA species revealed that eosinophils contained transcripts mainly encoding for the 121- and 165-amino-acid forms of VEGF. VEGF mRNA expression and VEGF release in cytokine-stimulated eosinophils were significantly reduced by treatment with a glucocorticosteroid, a protein-tyrosine kinase inhibitor, or a protein kinase C inhibitor. Cytokine activated eosinophils may be an important source of a vascular permeability factor, namely VEGF, thus contributing to tissue edema formation at sites of allergic inflammation. PMID- 9224212 TI - Inhibitory effect of heparin on serotonin-induced hyperplasia and hypertrophy of smooth muscle cells. AB - Serotonin (5-HT) produces both hyperplastic and hypertrophic effects on smooth muscle cell (SMC) in culture. Heparin is known to inhibit serum-induced hyperplasia of SMC but has not been previously tested on the stimulatory effect of 5-HT on SMC. Our present data show that at 24 h heparin inhibited by 50% the stimulation of 3H-thymidine incorporation into bovine pulmonary artery SMC and at 7 days totally reversed both cellular proliferation and enlargement of SMC produced by 1 microM 5-HT. Heparin failed to alter 5-HT uptake by SMC, but inhibited the stimulation of tyrosine phosphorylation of GTPase-activating protein, a proposed intermediate in the 5-HT stimulatory process. Thus heparin inhibits both hyperplastic and hypertrophic effects of 5-HT on SMC, perhaps through the inhibition of a phosphorylated intermediate protein. PMID- 9224213 TI - Distribution of receptors of collagen and globular domains of C1q in human lung fibroblasts. AB - Fibroblasts are the predominant cell type responsible for the synthesis of collagen and other matrix elements in normal and fibrotic lungs. We have previously reported that human lung fibroblasts are heterogeneous in C1q binding and that subpopulations differing in C1q binding can be isolated and subcultured. We have investigated the distribution of receptors for C1q-collagen domain (cC1q R) and globular domain (gC1q-R) in adult human lung fibroblasts. Fibroblasts were isolated from cultures of adult human lung explants in medium containing fresh- or heated plasma-derived human sera and separated by FACS-cell sorting into populations binding to C1q with high- (HF) and low- (LF) fluorescence. The cC1q-R was obtained from fibroblast membrane preparations by affinity chromatography through an anti-cC1q-R antibody column and its distribution was determined by Western analysis. The presence of gC1q-R was determined by immunoblots using an anti-gC1q-R antibody raised against a synthetic peptide. The results showed that a 54 kD protein crossreacting with anti-cC1q-R antibody was produced by LF cells, but it was barely detectable in HF cultures. Immunostaining with anti-cC1q-R antibody revealed that most of the cells in LF cultures were positive while the HF cells were negative. A 38 kD protein recognized by anti-gC1q-R antibody was produced by lung fibroblasts; however, no differences were detected in its distribution between LF and HF cultures. SDS-polyacrylamide gel electrophoresis of membrane proteins binding to an affinity column of C1q-globular fragment showed that the HF cultures contain a approximately 51 kD protein, which was a minor component in LF membranes. These data show that cC1q-R is expressed predominantly by a population of human lung fibroblasts, while the 38 kD gC1q-R is produced by all cells. Another 51 kD protein appears to be produced by a separate population of fibroblasts which does not express cC1q-R. Our results indicate that two lung fibroblast subtypes may be distinguished based on production of the 54 kD putative cC1q-R and another 51 kD protein which binds to C1q-globular domain. PMID- 9224214 TI - Alveolar macrophages in sarcoidosis coexpress high levels of CD86 (B7.2), CD40, and CD30L. AB - Alveolar macrophages (AM) from sarcoid patients have been shown to be good antigen presenting cells (APC) unlike normal AM which are usually ineffective. We demonstrate in ten consecutive sarcoid patients that most of their AM, unlike normal AM, do coexpress high levels of CD86, CD40, and CD30L, all known to be important for T-cell activation. CD80 is also slightly more expressed on sarcoid AM than on normal AM, but is detected on only 26 +/- 6% (mean +/- SEM) of sarcoid AM. A good correlation is present between the percentage of sarcoid AM expressing CD86 and CD40 or CD86 and CD30L. However, no correlation is found between the percentage of CD80 and CD86 positive AM in these same patients. Blocking antibodies against CD86 were able to reduce by more than 80% allogeneic T-cell proliferation induced by the AM of sarcoid patients. This study provides evidence that AM can, in pathologic states such as sarcoidosis, express functional costimulatory molecules for T-cell activation such as CD86, thought to be rather specific for more professional APC such as dendritic cells. PMID- 9224215 TI - Isolation and characterization of a peroxidase from the airway. AB - Sheep airway mucus can potently scavenge hydrogen peroxide, an important mediator of airway inflammation. Here, the scavenging activity was identified as a peroxidase produced by goblet cells of the airway epithelium and secreted into the airway lumen. Ovine airway peroxidase activity was purified approximately 100 fold from airway lavage fluid in two steps, using cation exchange and lectin affinity chromatography, yielding an apparently homogeneous 82-kD glycoprotein. Ovine airway peroxidase represented about 1% of the total protein in airway mucus and thus was an abundant enzyme in airway secretions. The absorbance spectrum of the purified peroxidase showed a major peak at 412 nm indicative of a hemoprotein. The ratio of A412/A280 of the purified enzyme was 0.86. The absorption spectrum of ovine airway peroxidase, its ability to oxidize halides, its sensitivity to inhibitors and its apparent molecular mass on sodium dodecyl sulfate gels showed that airway peroxidase was similar to lactoperoxidase but distinguished from myeloperoxidase, eosinophil peroxidase as well as from glutathione peroxidases. Based on these observations, ovine airway peroxidase is a newly isolated and abundant enzyme of airway mucus which may function to control reactive oxygen species in the airway and to prevent infection by catalyzing the formation of biocidal compounds. PMID- 9224216 TI - Identification of novel inducible genes in airway epithelium. AB - DNA differential display analysis (DD-PCR) was utilized to identify genes that are expressed in airway epithelium and are relevant to airway inflammation; cytokine-mediated induction of gene expression and inhibition of that induction by glucocorticoids were the criteria for selection. The IB3-1 cell line was cultured in the presence of tumor necrosis factor-alpha (TNF-alpha), dexamethasone, or dimethyl sulfoxide (DMSO) as a control, and analyzed via DD-PCR and Northern blot analyses. With this approach, two TNF-alpha-inducible and dexamethasone (DEX)-sensitive expressed sequence tags (EST8 and EST19) were identified. In IB3-1 cells, TNF-alpha increased messenger RNA (mRNA) expression of EST8 (34%, P < or = 0.005) and EST19 (41%, P < or = 0.01), whereas dexamethasone reduced this expression to resting levels. This pattern of mRNA expression was also observed in normal human bronchial epithelial cells (EST8: 21%, P < or = 0.009; EST19: 11%, P < or = 0.02) and in the basophil leukemia cell line KU812 (EST8: 34%, P < or = 0.01). Through basic local alignment search tool (BLAST) analysis, it was determined that these ESTs exhibited significant homology with the monomeric G protein rhoC (EST8: 100% homology, P = 1.6 x 10( 100)) and the UFO tyrosine kinase receptor (EST19: 86% homology, 5.3 x 10(-28). PMID- 9224218 TI - The association of microalbuminuria and mortality in non-insulin-dependent diabetes mellitus. A systematic overview of the literature. AB - OBJECTIVES: To critically analyze the literature linking microalbuminuria with total and cardiovascular mortality and cardiovascular morbidity in non-insulin dependent diabetes mellitus (NIDDM) and to quantify the risk. METHODS: A combination of retrieval techniques (MEDLINE, SCISEARCH, and handsearching published bibliographies) was used to find all relevant articles based on title and abstract and "Methods" sections. Unpublished data on albumin excretion rate were sought from large NIDDM cohort studies. RESULTS: A total of 264 citations were retrieved, of which 11 cohort studies were selected for inclusion in the overview, representing a total of 2138 patients followed up for a mean of 6.4 years. Patient age was similar across cohorts. Duration of NIDDM ranged from newly diagnosed to 13 years. The prevalence of microalbuminuria ranged from 20% to 36% in the 8 cohorts that excluded patients with clinical proteinuria. All studies reported either a trend or a significant association between microalbuminuria and total mortality or cardiovascular morbidity or mortality; the overall odds ratio for death was 2.4 (95% confidence interval, 1.8-3.1) and for cardiovascular morbidity or mortality, 2.0 (95% confidence interval, 1.4 2.7). We found no evidence of reporting bias. CONCLUSION: Microalbuminuria is a strong predictor of total and cardiovascular mortality and cardiovascular morbidity in patients with NIDDM. PMID- 9224217 TI - Quantitative RT-PCR measurement of cytochromes p450 1A1, 1B1, and 2B7, microsomal epoxide hydrolase, and NADPH oxidoreductase expression in lung cells of smokers and nonsmokers. AB - Bronchial epithelial cells (BEC) are the progenitors of bronchogenic carcinomas and are exposed to polycyclic aromatic hydrocarbon (PAH) procarcinogens through inhalation of combustion products. PAH are converted to carcinogenic molecules through a combination of monoxygenation by cytochrome p450 (CYP) enzymes in the presence of NADPH oxidoreductase (OR) and hydrolysis by microsomal epoxide hydrolase (mEH). In artificial systems, the relative expression of these genes determines whether carcinogenic or noncarcinogenic species are generated during metabolism. This relationship was explored in humans by using quantitative competitive reverse transcriptase polymerase chain reaction amplification to determine the range of expression of CYP1A1, CYP1B1, mEH, and NADPH OR in BEC recovered from 10 nonsmokers and 9 smokers. CYP2B7 expression was evaluated because, although little is known of its substrate specificity, it is expressed at high levels in human lung tissue. CYP1A1 and CYP1B1 were expressed in BEC at significantly different levels (P < 0.05) in the 9 smokers at 1.4 +/- 2.3 x 10(4) and 2.4 +/- 3.2 x 10(3) molecules/10(6) beta-actin molecules (mean +/- STD), respectively, but each was measurable in only one of the 10 nonsmokers. There was significant inter-individual variation (P < 0.05) in both CYP1A1 and CYP1B1 expression among the subjects for whom sufficient data were obtained. The inducibility of human BEC CYP1A1 gene by PAH exposure was confirmed in vitro by incubating cultured immortalized human BEC with beta-naphthoflavone and observing a > 6-fold induction of CYP1A1 after 24 h. In contrast to BEC, alveolar macrophages expressed CYP1A1 at low (30-70 molecules/10(6) beta-actin molecules) to unmeasurable levels in both smokers and nonsmokers. There was no significant difference in expression of mEH, CYP2B7, or NADPH OR in smokers compared with nonsmokers. The inter-individual variation in absolute and relative expression of PAH metabolism enzymes in BEC reported here supports the hypothesis that inter individual variation in ability to activate/inactivate inhaled PAH carcinogens accounts for at least some of the inter-individual variation in risk for bronchogenic carcinoma. PMID- 9224219 TI - Doxycycline revisited. AB - Although several new antibiotics have recently become available, in several clinical instances conventional antibiotics may be equally efficacious at a considerably lower cost. In today's era of cost containment, it is particularly relevant to revisit older, inexpensive antibiotics to reexplore their role in the face of the emergence of resistant microorganisms and competition from newer agents. Doxycycline is one such antibiotic. It is an inexpensive, broad-spectrum antimicrobial agent that remains the drug of first choice for several infections. In addition, it can be used for a variety of other indications. Adverse effects are infrequent and relatively minor. While interactions occur with several medications, none of these interactions has significant adverse consequences. PMID- 9224220 TI - The cost of dying of end-stage liver disease. AB - BACKGROUND: The high cost of liver transplantation is well known. The cost of dying of complications of end-stage liver disease (ESLD) without transplant, however, has not been well documented. METHODS: For a 5-year period (1991-1995), in 153 patients, mean inpatient hospital charges and length of stay were analyzed in 6 groups of patients: (1) patients admitted with the primary diagnosis of esophageal varices, (1a) the subset of group 1 patients who died on this admission, (2) patients admitted to the liver team who died of complications from ESLD, (3) patients who underwent transjugular intrahepatic portosystemic shunts, (4) patients who underwent surgical shunt for bleeding varices, and (5) patients who underwent liver transplantation. RESULTS: One hundred twenty-nine patients with esophageal varices were hospitalized 13.7 days with a mean charge of $30,980 for each of 202 admissions. Of these, 38 died after 24 days with a mean charge of $67,091. Seven patients admitted to the liver team died of complications of ESLD at $110,576 per admission. Transjugular intrahepatic portosystemic shunt was performed in 17 patients with a mean charge of $43,209. Six patients underwent surgical shunt for $53,994. Mean charge for 7 liver transplantations was $222,968. During the study period, 36.7% of all charges were for patients who died. CONCLUSIONS: It is difficult to estimate the total cost of ESLD; however, in evaluating inpatient costs, we see that it is expensive and significant amounts are spent on patients who die. Further study is necessary to determine which factors can optimize the cost of ESLD. PMID- 9224221 TI - Gout and risk for subsequent coronary heart disease. The Meharry-Hopkins Study. AB - BACKGROUND: Patients with gout are encountered frequently in clinical practice. Previous studies have suggested that hyperuricemia and gout may represent risk factors for coronary heart disease (CHD), the most common cause of death in American men. METHODS: Prospectively collected data from 2 longitudinal cohort studies of former medical students--371 black men in the Meharry Cohort Study and 1181 white men in the Johns Hopkins Precursors Study--were analyzed. The development of gout and of CHD was determined by physician self-report, and validated by using published criteria. The risk for CHD associated with gout was evaluated using Cox proportional hazards analysis. RESULTS: During a median follow-up of 30 years, there were 38 gout cases and 44 CHD events among the Meharry men, and 68 gout cases and 138 CHD events among the Hopkins men. Prior gout was not associated with an increased risk for incident CHD (relative risk = 1.20; 95% confidence interval, 0.37-3.92) among the Meharry men or among the Hopkins men (relative risk = 0.66; 95% confidence interval, 0.24-1.79). Multivariate analysis adjusted for known CHD risk factors did not alter these findings. CONCLUSION: These results, in black and white male physicians, do not suggest a role in men for targeting gout identification in the primary prevention of CHD. PMID- 9224222 TI - Eliminating unnecessary lactate dehydrogenase testing. A utilization review study and national survey. AB - BACKGROUND: Consensus recommendations call for the elimination of lactate dehydrogenase (LDH) tests from routine rule out myocardial infarction (ROMI) protocols. METHODS: We conducted a utilization review project in which we evaluated the institutional impact of removing LDH and LDH isoenzyme tests from our hospital diagnostic panel. We then conducted a scripted telephone survey of 100 US hospitals to assess the generalizability of this project. RESULTS: All our cardiology staff members supported this intervention. Lactate dehydrogenase isoenzyme test results did not add clinically useful data for any of 200 consecutive patients discharged with a diagnosis of acute myocardial infarction, and selective use of LDH isoenzyme testing in cases where it was clinically believed to be indicated cut costs 99% during the year after our intervention. Furthermore, our telephone survey demonstrated that 66% of US hospitals polled continue to test for LDH isoenzymes in every patient with possible myocardial infarction. CONCLUSIONS: Our results corroborate prior recommendations for the removal of LDH testing from the routine ROMI protocol. Such an intervention may be accomplished easily, with excellent staff acceptance and considerable savings. Most US hospitals continue to include LDH testing in their ROMI panels despite national guidelines recommending otherwise. PMID- 9224223 TI - The impact of alcohol-related diagnoses on pneumonia outcomes. AB - BACKGROUND: There is controversy regarding the role of alcoholism as a prognostic factor in hospitalized patients with pneumonia. OBJECTIVE: To assess the impact of alcohol abuse on hospitalization charges, length of hospital stay, intensive care unit use, and in-hospital mortality. METHODS: We studied a cohort of all adults hospitalized in 1992 in Massachusetts with a principal diagnosis of pneumonia, and all Massachusetts residents hospitalized for pneumonia in 6 bordering states. RESULTS: For the 23,198 pneumonia cases the mean total hospitalization charges were $9925, mean length of hospital stay was 9.6 days, 12% of the cases had intensive care unit stays, and 10% of the cases died during the hospitalization. In bivariate analyses, pneumonia cases with alcohol-related diagnoses had higher charges (mean, $11,232 vs $9877, P = .07), had shorter length of hospital stay (9.2 vs 9.6 days, P = .02), were more likely to experience an intensive care unit stay (19% vs 12%, P < .001), and had lower in hospital mortality (6.0% vs 10.2%, P < .001). Multivariable analyses adjusting for comorbidity, pneumonia etiology, and demographics revealed that for pneumonia cases with alcohol-related diagnoses, risk-adjusted hospital charges were $1293 higher (adjusted mean, $11,179 vs $9888, P < .001), length of hospital stay was 0.6 days longer (10.1 vs 9.5 days, P = .001), intensive care unit use was higher (18% vs 12%; adjusted odds ratio, 1.63; 95% confidence interval, 1.33-1.98), and mortality was no different (10% with or without an alcohol-related diagnosis). CONCLUSIONS: Having an alcohol-related diagnosis is associated with more use of intensive care, longer inpatient stays, and higher hospital charges. To understand resource utilization in cases of pneumonia, alcohol abuse is a comorbid factor that must be considered. PMID- 9224224 TI - Influence of age on symptoms at presentation in patients with community-acquired pneumonia. AB - BACKGROUND: Advanced age has become a well-recognized risk factor for death in patients with pneumonia. It may also be associated with reduced symptom reporting, raising the possibility that diagnosis and treatment may be delayed in older patients. OBJECTIVE: To evaluate the association between age and the presenting symptoms in patients with community-acquired pneumonia. METHODS: This study was conducted at inpatient and outpatient facilities at 3 university hospitals, 1 community hospital, and 1 staff-model health maintenance organization. Patients included adults (age > or = 18 years) with clinical and radiographic evidence of pneumonia, who were able to complete a baseline interview. The presence of 5 respiratory symptoms and 13 nonrespiratory symptoms were recorded during a baseline patient interview. A summary symptom score was computed as the total number of symptoms at presentation. RESULTS: The 1812 eligible study patients were categorized into 4 age groups: 18 through 44 years (43%), 45 through 64 years (25%), 65 through 74 years (17%), and 75 years or older (15%). For 17 of the 18 symptoms, there were significant decreases in reported prevalence with increasing age (P < .01). In a linear regression analysis, controlling for patient demographics, comorbidity, and severity of illness at presentation, older age remained associated with lower symptom scores (P < .001). CONCLUSIONS: Respiratory and nonrespiratory symptoms are less commonly reported by older patients with pneumonia, even after controlling for the increased comorbidity and illness severity in these older patients. Recognition of this phenomenon by clinicians and patients is essential given the increased mortality in elderly patients with pneumonia. PMID- 9224225 TI - Continuity of care and the use of breast and cervical cancer screening services in a multiethnic community. AB - OBJECTIVE: To examine how continuity of care affects the use of breast and cervical cancer screening in a multiethnic population. METHODS: All data came from a structured telephone survey of a population-based quota sample designed to determine the cancer prevention needs of multiethnic blacks and Hispanics in New York, NY, in 1992. The study included 1420 women of 7 racial/ethnic groups: US born blacks, English-speaking Caribbean-born blacks, Haitian blacks, and Puerto Rican, Dominican, Colombian, and Ecuadorian Hispanics. The main outcome measures were ever and recently having had a Papanicolaou smear, clinical breast examination (CBE), or mammogram. RESULTS: Among respondents who qualified for the survey on the basis of age and ethnicity, the refusal rate for completing the interview was 2.1%. Compared with women without a usual site of care, those with a usual site, but no regular clinician, were 1.56, 2.45 (P < or = .01), and 2.32 (P < or = .05) times as likely ever to have received a Papanicolaou smear, CBE, or mammogram, respectively and 1.84, 1.92 (P < or = .05), and 1.75 times as likely to have received a recent Papanicolaou smear, CBE, or mammogram, respectively. Compared with women without a usual site of care, women with a regular clinician at that usual site of care were 2.63 (P < or = .01), 2.83 (P < or = .01), and 2.30 (P < or = .05) times as likely ever to have received a Papanicolaou smear, CBE, or mammogram, and were 2.00 (P < or = .05), 2.65 (P < or = .01), and 1.40 times as likely to have recently received a Papanicolaou smear, CBE, or mammogram, respectively (adjusted odds ratios). For uninsured women, presence of a usual site of care was associated with increases in recent use of cancer screening for all screening tests. CONCLUSIONS: There is a linear trend in increasing breast and cervical cancer screening rates when one goes from having no usual source of care, to having a usual source, and to having a regular clinician at that usual source. Emphasis on continuity of care, especially on usual source of care, may help to bridge the gap in access to cancer prevention services faced by minority women. PMID- 9224226 TI - Effect of theophylline on erythrocytosis in chronic obstructive pulmonary disease. AB - BACKGROUND: Patients with chronic obstructive pulmonary disease (COPD) tend to develop secondary erythrocytosis to compensate for their chronic hypoxia. Theophylline has recently been shown to reduce hematocrit and erythropoietin blood levels in normal subjects and in patients with erythrocytosis after renal transplantation. OBJECTIVE: To determine whether theophylline may be used to lower the hematocrit in patients with COPD. METHODS: Two hundred four patients with COPD were studied retrospectively and 10 patients prospectively (8 starting treatment with the drug [group 1] and 2 who suspended its long-term use [group 2]) for the correlation between theophylline therapy and hematocrit and erythropoietin level. RESULTS: In the patients studied retrospectively, lower hematocrits were found in the theophylline-treated than in the untreated patients (0.43 +/- 0.006 vs 0.46 +/- 0.007, respectively; P < .002). Twelve untreated patients and 2 of those treated with theophylline had hematocrits above 52%. Oxygen saturation levels were similar in both groups, and exclusion of patients with oxygen saturation lower than 88% did not change the pattern, suggesting that the effect of theophylline could not be entirely explained by improved oxygen availability. Seven of the 8 patients studied prospectively in group 1 (P < .02) and the 2 patients in group 2 showed inverse correlations between hematocrits and theophylline administration. A similar pattern was observed with serum erythropoietin levels in 5 of 7 patients studied. The effects were reproducible on rechallenge in 3 of the 4 patients in group 1 and the 2 patients in group 2. CONCLUSIONS: Theophylline may have a beneficial effect in treatment and prevention of erythrocytosis in patients with COPD. PMID- 9224227 TI - Concerns and expectations in patients presenting with physical complaints. Frequency, physician perceptions and actions, and 2-week outcome. AB - BACKGROUND: Specific concerns and expectations may be a key reason that people with common physical complaints seek health care for their symptoms. OBJECTIVES: To determine the frequency of symptom-related patient concerns and expectations, physician perceptions and actions, and the relationship of these factors to patient satisfaction and symptom outcome. METHODS: This was a prospective cohort study of 328 adult outpatients presenting for evaluation of a physical complaint. The setting was a general medicine clinic in a teaching hospital. Measures included previsit patient questionnaire to identify symptom-related concerns and expectations; a postvisit physician questionnaire to determine physician perceptions and actions; and a 2-week follow-up patient questionnaire to assess symptom outcome and satisfaction with care. RESULTS: Pain of some type accounted for 55% of common symptoms, upper respiratory tract illnesses for 22%, and other physical complaints for 23%. Two thirds of patients were worried their symptom might represent a serious illness, 62% reported impairment in their usual activities, and 78%, 46%, and 41% hoped the physician would prescribe a medication, order a test, or provide a referral. Physicians often perceived symptoms as less serious or disabling and frequently did not order anticipated tests or referrals. While symptoms improved 78% of the time at 2-week follow-up, only 56% of patients were fully satisfied. Residual concerns and expectations were the strongest correlates of patient satisfaction. CONCLUSIONS: Improved recognition of symptom-related concerns and expectations might improve satisfaction with care in patients presenting with common physical complaints. PMID- 9224228 TI - Time trends of physician visits and treatment patterns of peptic ulcer disease in the United States. AB - BACKGROUND: In the last 4 decades, the prevalence rates of peptic ulcer disease and our understanding of its pathophysiological features underwent major changes. OBJECTIVE: To analyze how these trends affected physician visits and treatment of ulcer disease. METHODS: The National Diseases and Therapeutic Index of IMS America Ltd, Plymouth Meeting, Pa, was used as the data source. Survey data were obtained from a representative sample of US physicians 4 times per year during a 48-hour period and extrapolated to a national level. Physician visits for gastric, duodenal, and all peptic ulcers were expressed as rates per 100,000 living US population. RESULTS: Between 1958 and 1995, physician visits for duodenal ulcer showed a marked decline, while those for gastric ulcer remained largely unchanged. In 1995, 4 million patients visited a physician because of peptic ulcer, corresponding to a rate of 1500 per 100,000 US population. The predominant therapy changed from anticholinergics, tranquilizers, and antacids between 1958 and 1977 to histamine2 receptor antagonist from 1978 until 1988, which subsequently became replaced in part by sucralfate and proton pump inhibitors. In 1995, about 75% of ulcers were still treated primarily with antisecretory medications, and only 5% received antibiotic therapy. CONCLUSIONS: Peptic ulcer is still common, although duodenal ulcer rates continue to decrease. The historical trends of treatment regimens show a steady change between various medications. No therapeutic class dominated ulcer therapy for more than 20 years. This trend is likely to continue, particularly, in light of the small fraction currently treated by antibiotics to eradicate Helicobacter pylori. PMID- 9224229 TI - Risk factors for a medically inappropriate admission to a Department of Internal Medicine. AB - OBJECTIVE: To identify patient- and admission-related risk factors for a medically inappropriate admission to a department of internal medicine. METHODS: Cross-sectional study of a systematic sample of 500 admissions to the department of internal medicine of an urban teaching hospital. The appropriateness of each admission and reasons for inappropriate admissions were assessed using the Appropriateness Evaluation Protocol. Risk factors included the time (day of week and holidays) and manner (through emergency department or direct admission) of admission, patient age and sex, health status of patient and spouse, living arrangements, formal home care services, and informal support from family or friends. RESULTS: Overall, 76 (15.2%) hospital admissions were rated as medically inappropriate by the Appropriateness Evaluation Protocol. In multivariate analysis, the likelihood of an inappropriate admission was increased by better physical functioning of the patient (odds ratio [OR], 1.5; 95% confidence interval [CI], 1.1-2.1 [for 1 SD in Physical Functioning scores]), lower mental health status of the patient's spouse (OR, 2.6; 95% CI, 1.3-5.6), receipt of informal help from family or friends (OR, 3.3; 95% CI, 1.5-7.2), and hospitalization by one's physician (OR, 3.6; 95% CI, 1.7-7.5). Receiving formal adult home care was not associated with inappropriateness of hospitalization. CONCLUSIONS: Inappropriate admissions to internal medicine wards are determined by a mix of factors, including the patient's health and social environment. In addition, the private practitioners' discretionary ability to hospitalize their patients directly may also favor medically inappropriate admissions. PMID- 9224230 TI - Fulminant Pneumocystis carinii pneumonia in 4 patients with dermatomyositis. AB - Between 1989 and 1996, 4 cases of Pneumocystis carinii pneumonia (PCP) were observed in patients seronegative for the human immunodeficiency virus who were receiving corticosteroid therapy for dermatomyositis in our institution. These cases were considered unusual in light of the short delay of their onset after initiation of immunosuppressive therapy and their fulminant course: 3 of these patients died of PCP occurring during the first month of treatment with prednisone. In all 4 patients lymphopenia was observed before the initiation of corticosteroid treatment and low CD4 and CD8 cell counts were evident at the time of PCP. These observations support the view of an increase in both the severity and incidence of PCP in patients without human immunodeficiency virus infection and question the need for a primary prophylaxis in patients with connective tissue diseases receiving high-dose corticosteroid therapy. PMID- 9224231 TI - Underuse of warfarin in atrial fibrillation. PMID- 9224232 TI - Supraventricular distinctions, tachycardia, and panic disorder. PMID- 9224233 TI - An operation for severe grades of contracted or clawed toes. 1911. PMID- 9224234 TI - Rheumatologic view of the rheumatoid foot. AB - Rheumatoid arthritis is a common systemic disease that affects between 0.3% and 1.5% of the general population worldwide. In 1988, it was estimated by the National Arthritis Foundation that there were 4 to 6 million cases of rheumatoid arthritis in the United States. There is general agreement that the feet are a major source of pain and disability at some point in the course of the illness. The frequency of involvement of the feet among 1000 patients with rheumatoid arthritis studied by Vainio was 91% in females and 85% in males. The clinical features and pathogenesis of the rheumatoid foot and an approach to initial nonsurgical treatment will be discussed. PMID- 9224235 TI - Shoes and insoles for patients with rheumatoid foot disease. AB - Pedorthic devices (shoes, modifications, insoles) are alternatives to, and, complementary with, operative procedures for patients with rheumatoid foot disease. The practical use of insoles and shoesole rolls (rocker soles) to diminish pressures and shearing stresses, and to support load bearing, is described. Criteria for prescribing such devices are described. The requirements for orthopaedic shoes are listed and a special technique of using test shoes with transparent thermomoldable stiff plastic materials is introduced to reveal pressure spots in advance of finishing the definitive shoe. Custom made pedorthic devices can improve loading capacity and mobility effectively even in severely deformed feet, thus being an important part of medical treatment. PMID- 9224236 TI - Extraosseous manifestations of rheumatoid arthritis in the foot and ankle. AB - The immune mediated pathologic effects of rheumatoid arthritis on osteoarticular tissues are well delineated in the orthopaedic and medical literature. Less well explored are the extraosseous manifestations of rheumatoid arthritis. The rheumatoid disease process can affect virtually any organ system or tissue in the human body; from scleritis, arteritis, and splenomegaly to neuropathy, bursitis, and tendinopathy. The scope of this treatise is to define better for the clinician the extraosseous presentation of rheumatoid arthritis in the foot and ankle. PMID- 9224237 TI - Long-term followup of rheumatoid forefoot surgery. AB - Rheumatoid forefoot deformities were treated originally at the Rheumatism Foundation Hospital by metatarsal head resection (II-V) and resection of the base of the proximal phalanx of the great toe. Recurrent great toe deformity with pain in numerous cases led to a comparative series of arthrodesis of the first metatarsophalangeal joint with resection of lesser metatarsal heads. At an average followup period of 3 years, the clinical evaluation and patient assessments were slightly in favor of arthrodesis. However, the patients' evaluation at 14 years was slightly in favor of resection. Measured in the plane of the first metatarsophalangeal joint, the recommended fusion position is 15 degrees valgus and 30 degrees dorsiflexion (females) and 25 degrees dorsiflexion (males). The position of the fusion is critical for a successful surgical outcome. Although both surgical methods give good pain relief and patient satisfaction, there is a risk of reoperation in the long term. PMID- 9224238 TI - Surgery of the rheumatoid forefoot with special reference to the plantar approach. AB - The key to the understanding of rheumatoid forefoot deformities is the predominant joint involvement of the tibial side of the hindfoot, of the fibular tarsometatarsal joints, and the destruction especially of the metatarsal heads. Independent of these patterns, the predominant sources of pain and impaired function, and thus of surgical remedies, remains the metatarsophalangeal joints. The authors prefer forefoot arthroplasty consisting of a judicious resection of the metatarsal heads, plantar capsulorraphy, tenolysis and rerouting of the tendons, through a dorsomedial approach to the great toe basal joint, and through a transverse plantar approach with dermodesis (plastic resection of a plantar wedge of skin and subcutaneous tissue) for the lesser toes. This method yields excellent pain relief that does not deteriorate for at least 15 years. However, 1/2 of the nearly complete postoperative correction was lost again within 10 years, and barefoot walking again became more difficult. Reviewing the literature, one cannot detect a clearcut superiority of plantar versus dorsal approaches, nor of one surgical routine versus another. PMID- 9224239 TI - Arthroplasty of rheumatoid metatarsophalangeal joints. An outcome study. AB - Deformity of the forefoot is a common disabling problem in chronic rheumatoid arthritis. The most common deformifies are hallux valgus, cockup toes, and depressed metatarsal heads. Resection arthroplasty has been improved by adding reconstructive procedures, for example, medial capsular arthroplasty in the great toe and plantar plate arthroplasty of the lesser toes with longitudinal pinning of all toes for 4 weeks. Results were compared with a series of silicone double hinge implants without grommets in the great toe and resection of lesser metatarsal heads and pinning. Patients were evaluated by questionnaire to evaluate outcome. There was no significant difference in the two series of patients. Overall good results were 85% to 95% and slightly favored the group without implants. These results were equal to those reported in the literature for patients who underwent fusion of the great toe and resection arthroplasty of lesser toes. PMID- 9224240 TI - Rheumatoid arthritis. Hindfoot disease. AB - Rheumatoid arthritis frequently involves the hindfoot. It may produce a proliferative synovial hypertrophy around the tendons and a gradual destruction of the joints. Unfortunately, this part of the foot frequently is overlooked when caring for these patients who often have multiple areas of involvement giving pain and producing a deformity. Treatment consists of nonoperative measures such as proper footwear, physical modalities, and occasional injections of corticosteroids. It is important to preserve the function of the posterior tibial tendon and not allow the hindfoot to develop a valgus deformity. Arthrodesis of selected hindfoot joints provides relief of pain and prevents or corrects hindfoot deformity. It is critical to evaluate simultaneously the ankle joint in all these patients. PMID- 9224241 TI - Management of the rheumatoid hindfoot with special reference to talonavicular arthrodesis. AB - In the patient with rheumatoid arthritis, involvement of the hindfoot is common. If conservative management fails, surgical treatment should be considered before the development of a fixed deformity. In patients with isolated talonavicular disease, arthrodesis of this joint provides excellent pain relief and seems to prevent future deformity. PMID- 9224242 TI - Compression arthrodesis of the rheumatoid ankle and hindfoot. AB - The reported frequency of involvement of the rheumatoid ankle and hindfoot varies between 9% and 70%. Fusion of the ankle joint, the subtalar, talonavicular, or calcaneocuboidal joint (Chopart's joint) or all of them is the preferred method of treatment for severe rheumatoid involvement causing pain, instability, and/or severe deformity. Ankle arthroplasty is indicated rarely. Pantalar arthrodesis is performed more frequently than talonavicular fusion or ankle fusion. Reported rates of fusion after compression arthrodesis of the ankle joint vary from 65% to 90%, averaging 80% to 85%. Higher success rates of as high as 95% were obtained with internal lag screw fixation as proposed by Wagner. The result of various combinations of arthrodesis (n = 54) of the ankle joint, the subtalar joint, and Chopart's joint in 43 patients with rheumatoid arthritis operated on in a 10-year period from 1984 through 1993 are presented. In all cases internal fixation by lag screws according to Wagner was used with a modified lateral approach incorporating osteotomy of the distal fibula. The technique is described in detail. Solid fusion was obtained in 21% of the cases after 8 weeks, in 9% of the cases after 12 weeks, and in 92% of the cases after 16 weeks. In 8% (3 patients) revision because of delayed union or nonunion eventually led to bony fusion. Postoperative pain, walking capacity, gait, and the subjective outcome were assessed. Complications occurred in 16%, revision was performed in 11.6% of the cases; in all cases healing was obtained. Overall patient satisfaction was 93%. PMID- 9224243 TI - Use of grommets for flexible hinge implant arthroplasty of the great toe. AB - A double stemmed flexible hinge implant was designed in 1974 for use with the Mayo type resection arthroplasty of the first metatarsophalangeal joint. Press fit titanium grommets were developed in 1985 to protect the implant midsection from sharp bony edges and shearing forces that can lead to implant abrasion or fracture, and particulate synovitis. The study of 90 first metatarsophalangeal joint implant arthroplasties performed with encircling press fit titanium grommets showed favorable bone response around the implant stems and the bone to grommet interface, with absence of complications relating to particulate reactivity, implant or grommet fracture. These findings suggest that the grommets effectively protect the implant to bone interface, improve implant durability, and prevent particulate synovitis. PMID- 9224244 TI - Shoulder fusion for paralyzed upper limb. AB - Fusion of the shoulder joint after a brachial plexus injury is a well known procedure in cases of flail shoulder in combination with normal motor and sensory function in the band. However, in combination with modern orthoses to stabilize the elbow, fusion of the shoulder in cases of a totally flaccid and afunctional arm might be more beneficial. In a retrospective study the impact of shoulder fusion on daily abilities in a population with a completely flaccid arm caused by a brachial plexus injury was investigated. Compared with a similar population, consisting of 16 patients with an afunctional unstable shoulder, all 12 patients who underwent shoulder fusion proved to perform at a higher functional level. Shoulder fusion in combination with an elbow stabilizing orthosis for a completely flaccid upper limb is a beneficial procedure that leads to less disability and results in a better quality of life. PMID- 9224245 TI - Value of cervical spine radiographs as a screening tool. AB - A review of the reports of 848 cervical spine radiographs was done to assess the yield of useful and critical information in a group of patients without trauma. In 470 of these patients the clinical record also was reviewed; 54.2% of the radiographs were read as having degenerative change, 35% were read as normal, and 8.5% were read as being consistent with muscle spasm. The remaining 2.3% included diagnoses of anatomic or congenital variants, soft tissue calcification, or old compression fractures. There were no serious diagnoses such as acute fracture, dislocation, or neoplasm that, had they not been identified, would have put the patient in jeopardy. Thus, for most outpatients with nontraumatic symptoms of a nonspecific or nonlocalizing nature, the use of cervical spine radiographs as a screening tool is not justified. PMID- 9224246 TI - Periprosthetic bone density around fully hydroxyapatite coated femoral stem. AB - In this study, periprosthetic bone mineral density was measured at scheduled time intervals after surgery by dual energy x-ray absorptiometry in 21 patients to assess the history of bone density redistribution after femoral stem insertion. Measurements of changes in bone density with time were obtained for the regions of the greater trochanter, the lateral cortex, the tip, the medial cortex, and the calcar. In all regions, bone density decreased during the first 3 months after surgery; this was followed by a prolonged period of 18 to 30 months of bone gain, a subsequent period of steady state, and the final resumption of bone aging processes after the third postoperative year. The greatest loss was observed in the calcar region after 6 months (greater than 50%). The characteristic pattern of time related bone density changes obtained in this study may make it possible to compare other pathologic, design, or stiffness related patterns. This could have clinical relevance in the early diagnosis of pathologic processes and as a means of evaluating prosthetic designs. PMID- 9224247 TI - Wear particulate species and bone loss in failed total joint arthroplasties. AB - The aim of this study was to evaluate the relative contribution of polyethylene, metal, and polymethylmethacrylate (cement) particles to the overall bone loss in aseptic loosening. Twenty-four interface tissues with adjacent bone were obtained during 17 revision total joint arthroplasties (11 hips and six knees). Osteoclasts and macrophages were identified immunohistochemically on the bone surface. The length of the bone surface in contact with these cell types was measured and analyzed with reference to the particulate species present within the fibrous interface. The presence of abundant polyethylene particles significantly increased the proportion of the bone surface in contact with macrophages but did not have a significant influence on that of osteoclasts. Osteoclastic bone resorption was significantly more extensive in the presence of metal particles. In contrast, the presence of cement particles did not have a significant influence on macrophage or osteoclast coverage of the bone surface. These results highlight the significance of polyethylene particles in macrophage recruitment and subsequent osteolysis and suggest a different mechanism of bone loss related to metal, namely mediation through osteoclastic activities. The relative contribution of cement particles was negligible and needs reevaluation in light of evidence provided by others. PMID- 9224248 TI - Acetabular screw rings and surface treatment. AB - The Zweymuller truncated self tapered threaded ring is forged in pure titanium with a 3- to 5-mu grit blasted surface roughness. Among 167 consecutive primary total hip arthroplasties prospectively studied, two threaded components failed to achieve initial stability, with one requiring almost immediate revision. One hundred twenty-six hips have been fully documented with a 5-year minimum followup. The mean modified Harris hip score improved from 44.4 points preoperatively to 91 points at 7 years average followup (range, 60-121 months). Bone to implant gap rate decreased from 54.7% after surgery to 2.4% at last review, with only one detectable early migration that had secondary stable osseointegration. Annual linear wear of more than 0.2 mm per year could be detected in only one ceramic on polyethylene bearing surface with one corresponding femoral osteolysis granuloma. At the 9- to 10-year interval, the survivorship with definite loosening as a failure was 98.7%. Delayed loosening by failure of an initially stable reconstruction, progressive lucencies, liner disassociation, or worrisome osteolysis have not been observed. These midterm results are much better than those of former screw rings with smooth surface finishes. The main reason for the general failure and justified abandonment of threaded polished components may not be the screw in mechanism of their primary fixation, but the lack of an appropriate surface for bone ingrowth and osseointegration. PMID- 9224249 TI - Electromyographic analysis of muscle fatigue in anterior cruciate ligament deficient knees. AB - The aim of this study was to detect possible differences in muscle fatigue and recovery of knee flexor and extensor muscles in patients with a deficient anterior cruciate ligament compared with patients with a normal anterior cruciate ligament. Surface electromyography of 15 patients with anterior cruciate ligament deficiency was performed while the muscles were under 80% of maximum isometric contraction, and after 1, 2, 3, and 5 minutes of rest. During the first 60 seconds of contraction, all muscles recorded significantly decreased mean power frequency and increased amplitude. The rate of decrease of mean power frequency was significantly greater in the injured quadriceps and normal hamstrings. All muscles except two recovered to the initial mean power frequency level after 1 minute of rest. All but two muscles in the injured and normal limb recorded an overshoot of mean power frequency during the recovery phase. This overshoot phenomenon also was seen for some muscles in the amplitude analysis. The findings confirm the fatigue state in all the muscles, suggest recruitment of more Type II fibers as the muscle fatigues, and show the physiologic adaptation of the quadriceps and hamstrings to anterior cruciate ligament insufficiency. The current study indirectly shows a dissociation between low intramuscular pH and mean power frequency during the recovery phase. It also indirectly suggests that the atrophied thigh muscles have fiber type composition similar to that of the normal side. PMID- 9224250 TI - The natural course of arthrosis of the knee. AB - The aim of this study was to describe the course of untreated or conservatively treated arthrosis of the knee joint. Of 265 patients presenting with knee pain who had weightbearing radiographs taken between 1970 and 1973, 132 (33 men and 99 women) responded to and participated in a clinical followup 20 years later using the Hospital for Special Surgery score and new weightbearing radiographs. Arthrosis was defined as an equal to or greater than 50% joint space narrowing. In 75 of 132 knees of Ahlback Class 0 (57%) and 20 of 52 knees of Ahlback Class I (39%), the classification remained unchanged. The Hospital for Special Surgery score at followup was compared with the Ahlback classes in the 1970s for the 110 cases for which surgery was not done. A higher Ahlback class at the time of presentation was associated with lower functional score (Hospital for Special Surgery) 20 years later. In addition, in the contralateral knee a reduction of joint space may occur. In the arthrosis group with total joint space reduction or attrition there was a clinical and radiographic deterioration that would merit surgical intervention to avoid unnecessary loss of function. However, in knees with equal to or greater than 50% joint space reduction (Ahlback I) and pain, a considerable number (39%) did not deteriorate radiographically during a 20-year period, and 25% remain free of pain. It appears that the long-term prognosis of mild knee arthrosis is not necessarily poor. A substantial number of these patients will not have progression of the arthrosis. PMID- 9224251 TI - Laser Doppler flowmetry during open reduction for developmental dysplasia of the hip. AB - Laser Doppler flowmetry was used intraoperatively to monitor femoral head perfusion during open reduction of 13 congenital hip dislocations in 11 patients. Laser Doppler determinations ranged from 12 to 400 mV before reduction and 30 to 300 mV after reduction. Three patients had magnitude changes in excess of 50%. One had increased perfusion, and two had decreased blood flow. Avascular necrosis of the hip occurred in one patient that was not predicted by laser Doppler flowmetry. Femoral head perfusion measured 175 mV for the dislocated hip and 180 mV after reduction of the femoral head and completion of the pelvic osteotomy. The authors conclude that laser Doppler flowmetry is not a reliable method for monitoring femoral head perfusion during open reduction of the hip for developmental hip dysplasia. PMID- 9224254 TI - Recalcitrant nonunion. AB - One surgeon treated 13 patients with 14 long home fractures that remained ununited for 10 or more years (average, 16 years) and after an average of three prior surgeries. The clavicle was involved in two cases, the humerus in five (one proximal, three diaphyseal, and one distal intraarticular), the femoral diaphysis in three, and the tibial diaphysis in four. The patients were observed for an average of 54 months. All of the fractures healed, and every patient in the series regained functional use of the involved limb without reports of pain, instability, or persistent swelling related to the site of nonunion. Three patients had persistent leg length discrepancies, and five had substantial residual stiffness of one or both adjacent joints. This experience has shown that despite the longevity of the nonunions, healing can be achieved using the basic concepts of the creation of a stable skeletal fixation in the presence of a well vascularized environments with the addition of autogenous bone graft. By the same token, the duration of the nonunion will lead to soft tissue maladaptation and contracture that at times compromise successful restoration of limb length or adjacent articular mobility. PMID- 9224253 TI - Intracapsular hip pressure after femoral neck fracture. AB - A consecutive series of 34 patients with femoral neck fractures was included in a prospective study aimed at evaluating preoperative variations in intracapsular pressure after changes in hip position, hip traction, and aspiration of hemarthrosis and their influence on the development of femoral head necrosis. Patients were observed for 7 years after surgery. Before aspiration, the mean intracapsular pressure in the antalgic physiologic position was 44.4 mm Hg. There were no differences between displaced and undisplaced fractures. The pressure was a maximum (mean value, 124.8 mm Hg) with the hip in extension and inward rotation, this pressure being greater than the blood systolic pressure in most cases. Hip traction of 3 kg in the antalgic physiologic position was found to be highly effective in preventing any bone flow tamponade effect in displaced and undisplaced femoral neck fractures: the mean intracapsular pressure decreased to 28.5 mm Hg. Aspiration of the hemarthrosis induced a significant decrease in intracapsular pressure only in cases with impaired vascularity of the femoral head as measured by scintigraphy using 99mTc labeled methyldiphosphonate. Aspiration of the hemarthrosis therefore is indicated only in the above cases, although it is less effective than hip traction in the antalgic position. There was no significant correlation between intracapsular pressure and the scintigraphy ratio. Avascular necrosis of the femoral head was detected in six cases. Among these, five patients had an intracapsular pressure below their diastolic blood pressure. This could indicate that vascular damage related to the fracture could be an important cause of bone necrosis despite that blood supply can be decreased by a tamponade effect. PMID- 9224252 TI - The hip in children with cerebral palsy. Predicting the outcome of soft tissue surgery. AB - This study reviewed 56 hips in 37 children with cerebral palsy who had undergone an adductor tenotomy alone or in combination with an anterior obturator neurectomy. The mean review period was 5.3 years. At latest review, 25 of 30 (83%) hips with a preoperative migration percentage of less than 40% were reduced, but 20 of 26 (77%) hips with a preoperative migration percentage of 40% or more were subluxated or dislocated. Surgery was unsuccessful for 13 of 15 hips with an acetabular index of more than 27 degrees. Percutaneous adductor tenotomy alone was as effective as the combination of an open procedure with an anterior obturator neurectomy. The age at the time of surgery did not have a significant effect on the outcome. The preoperative migration percentage was the only significant predictor of outcome in this group of children. PMID- 9224255 TI - Modified lateral approach to the distal humerus for internal fixation. AB - Internal fixation of fractures of the most distal portion of the humeral shaft is problematic. A modified lateral approach was assessed to determine its role in the surgical management of these injuries. Posterior plating of the lateral column was performed in each case. Eight patients with eight fractures (seven acute, one nonunion) were treated. All fractures united. There were no complications. On the basis of these results, displaced fractures of the most distal portion of the humeral shaft can be managed successfully via a modified lateral approach to the distal humerus. PMID- 9224256 TI - Convergent dislocation of the elbow. AB - Proximal radioulnar translocation is an extremely rare injury. Only seven cases have been reported previously. The authors report the case of a 6-year-old girl whose translocation was undiagnosed for more than 2 weeks. Definitive treatment required open reduction and use of a radiocapitellar pin to stabilize the proximal radius. At the 2-year followup, the patient has no symptoms and has resumed full activity with nearly full range of motion. To ensure early diagnosis of rare types of elbow dislocations, it is important to recognize the lack of supination and disruption of the radiocapitellar relationship on radiographs. PMID- 9224257 TI - Difficulty in removal of certain intramedullary nails. AB - Intramedullary nailing is the most common treatment for displaced diaphyseal fractures of the femur and tibia. Gerhard Kuntscher introduced the technique of intramedullary nailing to clinical practice in the 1940s, and this method has been the focus of many authors with regard to indications, technique, complications, and outcome. The five cases presented here represent a complication not often reported in recent years: the difficulty in removing an intact intramedullary nail. Inspection of the interlocking nails in four of the cases presented reveals a specific design characteristic: the cross sectional design of the nail prevents the distal, unslotted end of the nail from being extracted from the medullary cavity. This problem is preventable by a change in nail design or the development of absorbable implants. PMID- 9224258 TI - Blood transfusion and bone allografts. Effect on infection and outcome. AB - Intraoperative and postoperative blood replacement have been implicated in increased rates of wound infection, decreased rates of renal allograft transplant rejection, and increased rates of local recurrence and metastasis of certain kinds of tumors, all presumably on the basis of some alteration in the immune system. Because patients who have bone allograft surgery for tumors often require transfusion and because the procedure is associated with a high rate of failure (20%), infection (9%-10%), and local recurrence (10% for high grade tumors), the effect of transfusion (range, 0-4750 ml) was studied for 264 patients who had proximal humeral, proximal or distal femoral resections, and massive cadaveric allografts but who did not have adjuvant chemotherapy or radiation. An attempt was made to statistically correlate the tumor and allograft outcome and rate of infection with patient age and gender, anatomic site, diagnosis, stage, type of graft, number of subsequent procedures, surgical margins, perioperative transfusions, blood loss, duration of operative procedures, and number of pregnancies. Of the variables studied, only blood loss, transfusion, and duration of surgery had an effect on outcome and, more specifically, on infection rate and time to union. No effect was observed on metastasis, recurrence, or the ultimate outcome of the procedure. PMID- 9224259 TI - Bone allografts are immunogenic and may preclude subsequent organ transplants. AB - The authors report a case of a 41-year-old woman with diabetes and chronic renal failure in whom antihuman leukocyte antigen antibodies developed after she received a frozen bone allograft that limited her access to organ donors. The patient had a chondrosarcoma of the right distal femur. A wide resection with segmental total knee arthroplasty was followed by a revision using a composite bone allograft prosthesis. After revision, broadly reactive lymphocytotoxic antibodies developed in the patient. The patient's panel reactive antibody level rose from 28% to a peak of 70%. Panel reactive antibody expresses the percentage of a panel of human leukocyte antigen type T lymphocytes from 40 individuals (representative of all human leukocyte antigen Class I histocompatibility antigens) to which antihuman leukocyte antigen Class I lymphocytotoxic antibodies have developed in the recipient as measured by the antiglobulin crossmatch method. The specificity of the patient's primary antibody is found in 45% of donors available in Illinois since 1988 (N = 1606). Because a positive crossmatch precludes kidney and pancreas transplantation, at least 45% of cadaver organ donors were excluded from use for this patient. This is an unusual case that focuses on the potential impact of bone allografts in patients who may need subsequent organ transplantation. PMID- 9224261 TI - Location of the extraforaminal lumbar nerve roots. An anatomic study. AB - Twelve cadavers were used to determine the anatomic location of the extraforaminal lumbar nerve root in the intertransverse space. After exposure of the transverse processes of the lumbar spine and the extraforaminal lumbar nerve roots, direct measurements, including the nerve root angle, nerve root diameter, distance to the superior facet, and the intertransverse space, were made bilaterally. The results showed that the extraforaminal nerve root angle and diameter and the distance between the superior facet and lateral limit of the nerve root consistently increased from cephalad to caudal. The largest dimension for height and width of the intertransverse space was found at the level of L3-4, and the smallest was found at the level of L5-S1. This information may be helpful in minimizing the incidence of injury to the lumbar nerve root during a posterolateral approach to lumbar disc. PMID- 9224260 TI - Osteochondral repair using perichondrial cells. A 1-year study in rabbits. AB - Articular cartilage repair remains a clinical and scientific challenge with increasing interest focused on the transplantation of chondrogenic cells. This study evaluated the repair response during a 1-year period after implantation of allogenic perichondrium cell polylactic acid composite grafts into 3.7 x 5 mm osteochondral defects drilled into the medial femoral condyles of 82 adult New Zealand White rabbits. The repair tissue was evaluated grossly, histologically, histomorphometrically, biochemically, and biomechanically at 6 weeks, 12 weeks, 6 months, and 1 year after implantation. After gross evaluation, cartilaginous material appeared to fill the defect in 70 experimental knees, for an overall repair frequency of 85%. The histomorphometric results and the histologic appearances were variable. None of the specimens were completely normal at 1 year. Only specimens with subchondral bone reformation displayed a definable cartilage appearing surface with chondrocytes surrounded by dense matrix. Subchondral bone reformation was inconsistent, reaching 50% at 1 year. Biochemically, the repair tissue matured during a 1-year period into a hyaline Type II collagen dominant tissue, whereas glycosaminoglycan content remained low at all time periods. The measured compressive properties of the repair tissue at 1 year were not significantly different from those of the contralateral knee that was not surgically treated. The treatment of osteochondral defects in the rabbit knee with allogenic perichondrium cell polylactic acid composite grafts yielded a high percentage of grossly successful repairs that showed inconsistent subchondral bone reformation. These results suggest that healthy subchondral bone is important to articular cartilage repair. They also highlight that a cartilaginous appearing tissue at gross inspection may not represent structurally normal articular cartilage. Continued multidisciplinary studies on the arthroplastic potential of rib perichondrial cells are needed before human studies, which rarely can extend beyond gross assessment of repair tissue appearance can be undertaken. PMID- 9224262 TI - Comparison of different methods used to inhibit physeal growth in a rabbit model. AB - A rabbit model was used to compare the rate, efficacy, and histologic appearance of physeal growth inhibition effected by Phemister epiphysiodesis, epiphyseal stapling, and percutaneous epiphysiodesis. Each technique led to an effective physeal closure, although the Phemister and staple methods produced more rapid deceleration of growth. A slower rate of physeal closure was seen after percutaneous epiphysiodesis, because this technique produced a gap in the bone that initially filled with fibrous tissue before forming bridges of trabecular bone leading to closure of the growth plate. Elevation of the periosteum alone produced an initial growth stimulation followed by early physiologic physeal closure. The amount of physis to ablate when doing a percutaneous epiphysiodesis is controversial. These results suggest that a percutaneous technique with limited physeal ablation, as used in the current study, effects slower rate of growth inhibition than that by the Phemister and staple techniques. A percutaneous technique that ablates a larger portion of the physis may be desirable to obtain more rapid growth inhibition. PMID- 9224263 TI - Evaluation of hip stability after simulated transverse acetabular fractures. AB - One of the major goals in managing acetabular fractures is the prevention of posttraumatic arthrosis. Unreduced fractures involving the weightbearing portion of the acetabulum may lead to posttraumatic arthrosis, whereas fractures outside this area portend a better prognosis. The purpose of this study was to help distinguish among fractures that require operative reduction, those that can be treated with traction, and those that require even less aggressive treatment. A model was developed to test hip stability after simulated transverse acetabular fractures. The results from this investigation suggest that transverse fractures with a roof arc angle of 90 degrees do not affect the weightbearing portion of the acetabulum. Fractures with a roof arc angle of 60 degrees begin to infringe on the weightbearing area, and those with roof arc angles of less than 60 degrees are clearly in the weightbearing region. Hip stability was significantly affected by the roof are angle and by the interaction of the roof arc angle and the angle of hip abduction or adduction. The data from the current study suggest that the area of the acetabulum considered to be weightbearing in transverse acetabular fractures may be more expansive than previously thought. The model developed may be used to investigate anterior and posterior column fractures. PMID- 9224264 TI - Patellofemoral joint kinematics and contact pressures in total knee arthroplasty. AB - Patellofemoral joint kinematics, contact areas, and contact pressures were measured concomitantly before and after total knee arthroplasty in 10 fresh frozen human cadaver knees using an Instron machine, a custom patellofemoral joint testing jig, axial bone markers, a continuous video digitizing system, and Fuji pressure sensitive film. The implant used in this study was the Kirschner Performance Knee System with an all polyethylene, domed patellar component. For all tests, the patella was aligned in its anatomically neutral position. Patellofemoral joint contact areas decreased as much as 19-fold after total knee arthroplasty. Mean patellofemoral joint contact pressures increased as much as 32 fold, and peak patellofemoral joint contact pressures increased as much as 22 fold after total knee arthroplasty. No statistically significant differences between preoperative and postoperative specimens were observed with respect to the patellofemoral, patellotibial, or patellar tilt angles from 30 degrees to 120 degrees knee flexion. Thus, the elevated patellofemoral joint contact pressures observed after total knee arthroplasty in vitro are not a primary consequence of iatrogenically altered patellofemoral kinematics. PMID- 9224265 TI - Stimulation of systemic bone formation induced by experimental blood loss. AB - Direct physical injury to bone marrow is associated with a systemic osteogenic response. However, blood loss, a condition that stimulates hemopoietic stem cells, also may activate osteoprogenitor cells in the bone marrow. To determine if bleeding induces a systemic osteogenic response, the mineral appositional rates and osteoblast numbers were determined in the bones of rats that were subjected to controlled cardiac bleeding and compared with those of rats subjected to ablation of their tibial bone marrow. In addition, a study of the kinetics of the osteogenic responses during the first 10 days after operative treatment was performed by quantitating the serum levels of biochemical indices known to be associated with systemic bone formation. The results showed that animals that sustained acute blood loss (1% or 3% body weight) or injury to their tibial bone marrow had statistically significant increases in mineral appositional rate, osteoblast number, and serum levels of osteogenic growth peptide. The kinetics studies showed that osteogenic growth peptide levels peaked on the tenth postoperative day and declined sharply thereafter. An enhancement of serum osteocalcin activity occurred only on the second postoperative day, was increased in all experimental groups when compared with untreated control animals, but immediately declined to baseline levels. Alkaline phosphatase activities increased in the experimental groups, peaking on Day 10 after tibial bone marrow ablation and on Day 12 in the group that underwent bleeding. These findings suggest that bleeding alone, independent of any skeletal trauma, may evoke a systemic osteogenic response. This response is similar in its timing and magnitude to that which has been shown to follow direct physical injury to bone marrow. The observation that systemic bone formation follows bone marrow activation induced by two different stimuli suggests that these responses may be mediated by common regulatory mechanisms. The ability to trigger or control these responses may form the basis for future therapeutic strategies to enhance bone formation. PMID- 9224266 TI - Lateral leg pain in a 26-year-old woman. PMID- 9224267 TI - How best to treat acute or unstable slipped capital femoral epiphysis with great interest. PMID- 9224268 TI - The historical development and clinical results on metal on metal total hip systems. PMID- 9224269 TI - Influence of surgical approach on lateral retinacular releases in total knee arthroplasty. PMID- 9224270 TI - Isolation of bone from muscles prevents the development of experimental callus like heterotopic bone. PMID- 9224271 TI - Anterior instability of the knee. PMID- 9224272 TI - No more 'squint' please. PMID- 9224273 TI - Hyperglycemic cytosolic reductive stress 'pseudohypoxia': implications for diabetic retinopathy. PMID- 9224274 TI - Retinoic acid. A key molecule for eye and photoreceptor development. PMID- 9224275 TI - Catoptric properties of eyes with misaligned surfaces studied by exact ray tracing. AB - PURPOSE: Historically, Purkinje images have been used to calculate ocular surface curvature and misalignment. The purpose of this report is to introduce an exact ray-tracing program that can be used to examine the influence of ocular component variations on the size and position of Purkinje images I, III, and IV. METHODS: Ray tracing was carried out on Le Grand's four-surfaced schematic eye to demonstrate the main features of the program. Any location may be chosen for the point light source and the observer. Ocular component dimensions, eye rotation, and crystalline lens decentering and rotation are fully adjustable. The program computes the coordinates of the Purkinje images in three-dimensional space from the point of view of the observer. It also offers the option of exhibiting Purkinje images seen through standard and telecentric imaging devices. Both options yield Purkinje image positions in relation to the rotating center of the limbus, observed clinically. RESULTS: The resulting program compared favorably with current ray-tracing software and with data from the literature. CONCLUSIONS: The program has many research and teaching applications. For example, our requirement was to calculate a series of linear coefficients that closely approximate Purkinje-image behavior in any given eye. These coefficients may be used to measure misalignment of intraocular components in phakic and pseudophakic eyes. PMID- 9224276 TI - Measurement of ultraviolet radiation at the surface of the eye. AB - PURPOSE: A new method for the measurement of ultraviolet radiation that reaches the surface of the eye is described. METHODS: The technique uses contact lenses produced from the ultraviolet-sensitive plastic polysulfone. Two types of polysulfone contact lenses (9 mm and 12 mm in diameter) were manufactured from a polysulfone rod. The 9-mm polysulfone contact lens could be calibrated and used to determine the ocular-to-ambient exposure ratio in a fashion similar to polysulfone film badges. The 12-mm polysulfone contact lens was designed as a "piggy-back" lens and required a larger diameter polymethlylmethacrylate carrier lens to fit the eye adequately. A method of in vivo stabilization was developed to minimize lens rotation. RESULTS: During four wearing trials, the ratio of ocular-to-ambient ultraviolet exposure ranged from 4% to 23%. CONCLUSIONS: Contact lenses manufactured from polysulfone offer the potential to study the exposure of the eye to ultraviolet radiation. The smaller diameter lens can measure an average ocular exposure, whereas the larger, stabilized, piggy-back design may allow regional dose assessment across the entire lens surface. PMID- 9224277 TI - Improvement in Vernier acuity in adults with amblyopia. Practice makes better. AB - PURPOSE: To determine the nature and limits of visual improvement through repetitive practice in human adults with naturally occurring amblyopia. METHODS: A key measure the authors used was a psychophysical estimate of Vernier acuity; persons with amblyopia have marked deficits in Vernier acuity that are highly correlated with their loss of Snellen acuity. The experiment consisted of three phases: pretraining measurements of Vernier acuity and a second task (either line detection thresholds or Snellen acuity) in each eye with the lines at two orientations; a training phase in which observers repetitively trained on the Vernier task at a specific line orientation until each had completed 4000 to 5000 trials; and posttraining measurements (identical to those in the first phase). Two groups of amblyopic observers were tested: novice observers (n = 6), who had no experience in making psychophysical judgments with their amblyopic eyes, and experienced observers (n = 5), who had previous experience in making Vernier judgments with their amblyopic eyes (with the lines at a different orientation) using the signal-detection methodology. RESULTS: The authors found that strong and significant improvement in Vernier acuity occurs in the trained orientation in all observers. Learning was generally strongest at the trained orientation but may partially have been transferred to other orientations (n = 4). Significant learning was transferred partially to the other eye (at the trained orientation) in two observers with anisometropic amblyopia. Improvement in Vernier acuity did not transfer to an untrained detection task. In two observers, the improvement in Vernier acuity was accompanied by a commensurate improvement in Snellen acuity. CONCLUSIONS: Some adults with amblyopia retain a significant degree of neural plasticity. Although several observers (primarily novices) showed evidence of generalized learning, several amblyopic patients showed evidence for improvement that was orientation and task specific. In this latter group of observers, the improvement appeared to reflect alterations that were, at least in part, in early neural processes that were orientation specific and were localized beyond the site of convergence of the two eyes. PMID- 9224278 TI - Integration of a sensory component into the accommodation model reveals differences between emmetropia and late-onset myopia. AB - PURPOSE: To evaluate the differences in accommodative function between subjects with emmetropia and those with late-onset myopia (LOM). METHODS: This study suggests a modified model of static accommodation, in which an accommodative sensory gain as a linear operator is added to simulate the sensory part of the system. Results derived from the model show that the sensory part not only affects the slope of the accommodative response function but also increases the system's effective threshold (ET) to the blur signal. This method expands the utility of using the control model to evaluate accommodation behavior. Thirteen emmetropic and 10 LOM subjects participated in this study. The subject's accommodative responses to one-, two-, three-, and four-diopter stimuli were measured by the Canon R-1 optometer, and the differences in dark focus, the slope of the accommodative response function, and the ET were compared between the emmetropic and the LOM subjects. RESULTS: The results show that although the dark focus values and the slopes of the accommodative response function are not significantly different in emmetropia and LOM, the ETs are significantly different. CONCLUSIONS: The higher ET found among subjects with LOM suggests that either the blur (or the error) signal is degraded significantly in the sensory part of the system, the dead space as an internal threshold of the system is high, or both factors are important. On the basis of further analysis of the data, we speculate that the sensory system in LOM subjects was less sensitive to blur than that of the emmetropic subjects. PMID- 9224279 TI - Factors affecting the pores of the inner wall endothelium of Schlemm's canal. AB - PURPOSE: A linear relationship between the density of pores in the inner wall of Schlemm's canal and aqueous outflow facility has been reported previously in a study in which investigators examined only eyes fixed at constant pressure, so that fixative flow rates differed from eye to eye. Because pores may form as a function of flow rate, the purpose in the current study was to verify the previous findings, using constant flow perfusions. METHODS: Outflow facility was measured in enucleated human eyes. Eyes were fixed under either constant flow or constant pressure conditions, microdissected to expose the inner wall of Schlemm's canal, and prepared for scanning electron microscopy. The density and diameter of pores in the inner wall were measured. RESULTS: Statistical analysis showed no correlation between outflow facility and either the density or the diameter of pores. Pore density decreased significantly during the hours after death. Examining only eyes for which experimentation was started within 20 hours of death, we found that pore density increased significantly with the volume of fixative that had been perfused through the outflow pathway. CONCLUSIONS: The correlation found by Allingham et al between outflow facility and pore density in the inner wall endothelium was not confirmed. However, the relationship between pore density and volume of fixative perfused is consistent with and may be responsible for the finding in the previous study. Because fixation conditions can influence the apparent pore density in the inner wall endothelium significantly, the conclusion reached previously, that pores contribute only 10% of the aqueous outflow resistance, may require reevaluation. PMID- 9224280 TI - Nonsulfhydryl-reactive phenoxyacetic acids increase aqueous humor outflow facility. AB - PURPOSE: The phenoxyacetic acid, ethacrynic acid (ECA), has potential use in glaucoma therapy because it acts to increase aqueous outflow in vivo and in vitro. In human trabecular meshwork (HTM) cell culture, ECA acts to change cell shape and attachment, effects that have been correlated with microtubule (MT) alterations and chemical sulfhydryl (SH) reactivity. To further explore these actions, we evaluated two non-SH reactive phenoxyacetic acids, inadcrinone and ticrynafen, and the MT-disrupting drug vinblastine. METHODS: Excised bovine and porcine eyes were perfused and outflow facility measured. Calf pulmonary artery endothelial and HTM cells were grown in culture and cytoskeletal effects evaluated after drug treatment. RESULTS: Indacrinone, ticrynafen, and vinblastine all caused an increase in outflow facility. In contrast with ECA, the outflow effects of indacrinone and ticrynafen were not blocked by excess cysteine. Although indacrinone and ticrynafen produced changes in cell shape in vitro, the beta-tubulin staining pattern of treated cells was not altered. Vinblastine caused cell shape change and the expected MT disruption. CONCLUSIONS: Phenoxyacetic acids can increase aqueous outflow facility and alter HTM cell shape and attachment in vitro by a non-SH, non-MT mechanism (which is probably shared also by ECA). These findings suggest the possibility of a broader class of glaucoma drugs that may be directed at the HTM. An understanding of the cellular target for these drugs has implications both for potential glaucoma therapy and for the cytoskeletal mechanisms involved in normal outflow function. PMID- 9224281 TI - Pseudomonas deficient in protease IV has significantly reduced corneal virulence. AB - PURPOSE: The role of protease IV in the pathogenesis of Pseudomonas aeruginosa keratitis was investigated by comparing a mutant strain completely deficient in protease IV activity with its protease IV activity-producing parent. METHODS: A protease IV-deficient Pseudomonas strain PA103-29::Tn9 was generated by mutagenesis of strain PA103-29, which produces protease IV, through transposon insertion. Protease IV activity was determined by a casein agar assay, zymography, and cleavage of the chromogenic substrate, Chromozym PL. Corneal virulence was evaluated by slit lamp examination and bacterial cultures in both a rabbit intrastromal model and a mouse topical model of keratitis. RESULTS: The protease IV-deficient strain PA103-29::Tn9 had significantly reduced corneal virulence relative to its parent strain PA103-29 in both a rabbit intrastromal model and a mouse topical model of infection. In the rabbit model, ocular damage (slit lamp examination score) mediated by the parent strain was severe at 32 hours after infection, whereas damage mediated by the mutant was minimal at both 32 and 55 hours after infection. This difference in virulence was not a result of differences in growth in vivo, because both strains grew equally. In the mouse model, eyes inoculated with the protease IV-producing parent strain had significant corneal damage as early as 24 hours after infection, whereas the protease IV-deficient mutant strain produced no significant corneal damage during 6 days of infection. CONCLUSIONS: The ability to produce active protease IV was the determining factor in the severity of corneal virulence. Protease IV appears to mediate corneal virulence and should be considered as a target in the development of medications designed to minimize corneal damage during Pseudomonas keratitis. PMID- 9224282 TI - Hepatocyte growth factor, keratinocyte growth factor, and other growth factor receptor systems in the lens. AB - PURPOSE: To examine the expression and function of hepatocyte growth factor (HGF), keratinocyte growth factor (KGF), epidermal growth factor (EGF) and other growth factor-cytokine-receptor systems in lens epithelial cells. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis were used to examine the expression of messenger RNAs in primary cultured rabbit and human lens cells and in ex vivo rabbit lens tissue. Protein expression and the effect of HGF and KGF on crystallin expression in lens epithelial cells were evaluated by immunoprecipitation and Western blot analysis. The effect of exogenous HGF, KGF, and EGF and of the coculture of lens epithelial cells with corneal endothelial cells on the proliferation of rabbit lens cells in a Transwell system was determined by cell counting. RESULTS: Messenger RNAs and proteins of HGF and KGF were expressed in primary rabbit lens epithelial cells and in ex vivo rabbit lens epithelial tissue. Human lens cells also expressed the mRNAs. Other growth factors and receptor messenger RNAs were also expressed. Hepatocyte and keratinocyte growth factors, and coculture with corneal endothelial cells stimulated proliferation of rabbit lens epithelial cells. In first-passage rabbit lens cells, HGF, KGF, and EGF increased the expression of alpha and beta crystallins. CONCLUSIONS: Hepatocyte and keratinocyte growth factor-receptor systems are expressed in lens cells. HGF and KGF are not expressed in epithelial cells in such tissues as skin, cornea, and lacrimal gland in which fibroblastic and epithelial cells interact in the formation of an organ. Expression of these growth factors in the lens may have evolved because the lens cells are relatively isolated within the anterior chamber of the eye. Our results suggest, however, that growth factors released by the corneal endothelium also could modulate lens functions (aquecrine interactions). PMID- 9224283 TI - Expression of macrophage migration inhibitory factor in corneal wound healing in rats. AB - PURPOSE: The study was conducted to evaluate the expression of macrophage migration inhibitory factor (MIF) during penetrating corneal injury. METHOD: A penetrating linear incision (2 mm) was made in the center of the right cornea with a razor blade. The expression of MIF in the lacerated eye and in the contralateral eye was examined by immunohistochemistry at 3, 6, 24, 48, and 72 hours after injury. Concentrations of MIF in the aqueous humor of the injured and contralateral eyes were also measured by enzyme-linked immunosorbent assay. The expression of MIF messenger RNA (mRNA) in the injured cornea was quantified by reverse transcription-polymerase chain reaction and subsequent Southern blot analysis. RESULTS: Positive migration inhibitory factor staining was observed in the basal cells of epithelial and endothelial cells of the normal rat cornea. The positive staining of the central corneal epithelium diminished at 3 hours after injury. At 6 hours after injury, positive MIF staining reappeared on the basal cells of the injured area, whereas the staining of the contralateral eye remained unchanged. Enzyme-linked immunosorbent assay of the aqueous humor revealed that the MIF concentration was elevated in both the injured and the contralateral eyes. The maximum concentration of aqueous MIF was observed at 6 hours after injury in both eyes. Reverse transcription-polymerase chain reaction and Southern blot analysis revealed that MIF-mRNA expression in the injured cornea increased from 6 to 48 hours after injury. CONCLUSION: The results of immunohistochemistry suggest the possibility that MIF is released from the corneal epithelial cells of the injured eye within 3 hours. Conversely, the MIF-mRNA level of the injured cornea is increased from 6 to 48 hours after injury and then diminished. In addition, unilateral corneal injury induces bilateral upregulation of MIF in the aqueous humor. PMID- 9224284 TI - Oxygen modulation of guanylate cyclase-mediated retinal pericyte relaxations with 3-morpholino-sydnonimine and atrial natriuretic peptide. AB - PURPOSE: This study explores at which level of the guanylate cyclase pathway oxygen modulates retinal pericyte relaxation induced by nitric oxide (NO). METHODS: Bovine retinal microvascular pericytes were grown on silicone. On silicone, pericyte contractile tone induces wrinkles. Drug-induced relaxation was quantified as a reduced number of wrinkles after exposure to 3-morpholino sydnonimine (SIN-1) or atrial natriuretic peptide (ANP) in the absence or in the presence of either 0.3 microM methylene blue (MB), a guanylate cyclase inhibitor, or 10 microM hemoglobin, a NO scavenger; and under 100% oxygen (hyperoxia), ambient air (normoxia), or 100% nitrogen (hypoxia). RESULTS: Pericytes were relaxed with SIN-1 and ANP in a concentration-dependent manner (EC50: 0.1 microM and 0.01 microM, respectively). Relaxations induced by SIN-1 or ANP were inhibited (P < 0.001) by MB, whereas hemoglobin inhibited only SIN-1 relaxations (P < 0.001). Relaxations induced by SIN-1, but not by ANP were increased (P < 0.001) under hypoxia and decreased (P = 0.002) under hyperoxia. CONCLUSIONS: SIN 1 and ANP relax pericytes through the activation of guanylate cyclase (inhibited by MB), but only SIN-1 through an extracellular release of NO (inhibited by hemoglobin). That oxygen only modulates pericyte relaxations induced by SIN-1 (NO mediated) but not those induced by ANP suggests that an interaction between oxygen and NO might participate in the capillary network's blood-flow modulation according to local tissue oxygen tension. PMID- 9224285 TI - An assessment of rat photoreceptor sensitivity to mitochondrial blockade. AB - PURPOSE: To report results of functional, biochemical and structural studies of photoreceptor mitochondria in isolated rat retinas under conditions of mitochondrial inhibition. METHODS: Dark-adapted rat retinas were incubated in a modified Ringer's bicarbonate medium under aerobic and anaerobic conditions. Several different procedures were used to inhibit mitochondrial function; N2, 0.01 mM antimycin A, and 1 and 10 mM potassium cyanide (KCN). Measurements were made of lactic acid production, retinal adenosine triphosphate (ATP) content, and receptor potentials. Morphology of the inner segment mitochondria was examined by electron microscopy. RESULTS: In the presence of N2, 0.01 mM antimycin, or 1 mM KCN, lactic acid production was linear throughout the 60- minute period; and the rate was similar for each condition. Retinal ATP content and the amplitude of the receptor potential were also maintained at high levels after short-term incubations with either N2, antimycin A, or 1 mM KCN. In contrast, use of 10 mM KCN produced an entirely different set of results. These effects were studied both at the alkaline pH (8.9) found when this concentration of KCN was simply added to bicarbonate-buffered media and at the normal pH (after readjustment) of 7.4. With 10 mM KCN (pH 8.9), retinal lactate production was severely depressed, retinal ATP content was nearly depleted within 5 to 10 minutes, and the amplitude of the receptor potential rapidly declined to a low level. The deleterious effects of 10 mM KCN on these parameters were lessened to varying degrees when pH was readjusted to 7.4. Electron microscopic observations of rat rod inner segments indicated generally excellent survival of these organelles after incubation with either N2, antimycin A, or 1 mM KCN in comparison with their appearance under oxygenated conditions. However, the inner segments were significantly disrupted after incubation of retinas with 10 mM KCN. CONCLUSIONS: Findings suggest that the loss of the receptor potential and depletion of ATP observed with minutes after exposing isolated rat retinas to media containing 10 mM KCN results from the inhibition of both respiration and glycolysis by this high concentration of KCN. In contrast, when conditions are chosen so that only respiration is impaired (as with N2, antimycin A, or 1 mM KCN) photoreceptor cells are resistant to short-term episodes of mitochondrial inhibition, principally because the upregulation of glycolysis generates sufficient ATP to compensate reasonably well for the loss in mitochondrially produced ATP. PMID- 9224286 TI - Characterization of N5-methyltetrahydrofolate uptake in cultured human retinal pigment epithelial cells. AB - PURPOSE: To determine the identity of the transport process that is responsible for the uptake of N5-methyltetrahydrofolate, the predominant form of folate in blood, into cultured human retinal pigment epithelial cells. METHODS: Human retinal pigment epithelial cells were cultured on an impermeable plastic support, and the characteristics of the uptake of radiolabeled N5-methyltetrahydrofolate into the cells were investigated. The expression of the folate receptor and the reduced-folate transporter in these cells was evaluated by functional assays and by Northern blot analysis. In addition, the characteristics of N5 methyltetrahydrofolate uptake in these cells were compared with those in folate transport-defective human breast cancer cells that were manipulated to express functionally by transfection the cloned human folate receptor and the human reduced-folate transporter. RESULTS: Transport of N5-methyltetrahydrofolate into these cells occurred by a single saturable process with a Michaelis-Menten constant of 0.13 +/- 0.01 microM. The process was specific for such reduced folates as N5-methyltetrahydrofolate and N5-formyltetrahydrofolate. Nonreduced folate interacted with this transport process only weakly. The transport process was inhibited by anion transport inhibitors. Results of Northern blot analysis indicated the presence of transcripts specific for the reduced-folate transporter in these cells. These cells expressed very small amounts of the folate receptor evidenced from the binding of folate and from the detectable presence of folate receptor-specific transcript, but there was no evidence for participation of the receptor in the observed transport of N5-methyltetrahydrofolate. There was also no evidence in these cells for expression of the folate transporter that prefers nonreduced folate as a substrate. CONCLUSIONS: Transport of N5 methyltetrahydrofolate in human retinal pigment epithelial cells occurs exclusively through the reduced-folate transporter. The folate receptor is expressed at negligible levels in these cells and does not participate in the observed transport. Because the cells were cultured on impermeable supports, making the basolateral membrane inaccessible for transport measurements, it is speculated that the observed findings are related to the transport function of the apical membrane of these polarized cells. PMID- 9224287 TI - Vitronectin is responsible for serum-stimulated uptake of rod outer segments by cultured retinal pigment epithelial cells. AB - PURPOSE: To examine whether the vitronectin (VN) in serum is responsible for the serum stimulation of phagocytosis in the rod outer segment (ROS) by cultured retinal pigment epithelial (RPE) cells. METHODS: Vitronectin was removed from fetal bovine serum by heparin-agarose affinity chromatography. Concentrations in normal and depleted serum were determined by enzyme-linked immunosorbent assay, using a polyclonal antibody against bovine VN and commercially prepared human VN as a standard. A monoclonal antibody against human alpha v beta 5 was used in localization and in blocking experiments. Rod outer segment phagocytosis was measured using a flow cytometric assay. RESULTS: Affinity chromatography removed 95% of the VN from serum as determined by enzyme-linked immunosorbent assay. Vitronectin-depleted serum did not stimulate ROS phagocytosis by RPE cells. Commercially prepared VN added to serum-free medium stimulated ROS phagocytosis in a dose-dependent manner. Pretreatment of RPE cells with an antibody against alpha v beta 5, an integrin receptor for VN, had no effect on phagocytosis in the absence of serum but completely blocked the serum stimulation of ROS phagocytosis. Antibody against alpha v beta 5 demonstrated a variable labeling pattern on the cultured RPE cell surface with morphologically distinct cell clusters exhibiting less labeling. Those cell clusters exhibiting less receptor labeling also showed less uptake of fluorescent-labeled ROS. CONCLUSIONS: Vitronectin is the component responsible for serum stimulation of ROS uptake, and this uptake appears to be mediated by an alpha v beta 5 integrin. Although clearly important in vitro, a role for VN in ROS uptake by RPE cells in situ remains to be determined. PMID- 9224288 TI - Effect of increasing glucose concentrations and protein phosphorylation on intercellular communication in cultured rat retinal pigment epithelial cells. AB - PURPOSE: The intercellular communication between cultured rat retinal pigment epithelial (RPE) cells grown in increasing glucose concentrations or after modulation of the protein kinase C-induced protein phosphorylation was investigated by studying the conduction of the [Ca2+]i wave elicited by mechanical stimulation and by analyzing the fluorescence recovery after photobleaching (FRAP). METHODS: Subconfluent monolayers of RPE cells isolated from neonatal Long Evans rats were cultured in growth medium with various glucose levels and analyzed using the fluorescent dye fluo-3 for measurements of intracellular Ca2+ after mechanical stimulation and using 6-carboxyfluorescein diacetate to investigate the intercellular communication with FRAP. RESULTS: Mechanical stimulation in 5 or 12 mM glucose resulted in a Ca2+ wave that spread centrifugally through the neighboring cells. An inhibition of the propagation of this wave, similar to that induced by halothane, could be observed in cells grown for 72 hours in 14-mM or higher concentrations of glucose. This inhibitory effect was not caused by a hyperosmotic effect, in that results of experiments on cells cultured in growth medium supplemented with mannitol instead of glucose did not differ from those of experiments in the control medium. Activation of protein kinase C by incubation of the cells for 30 minutes with phorbol 12-myristate 13 acetate (PMA) resulted in a strong inhibition of [Ca2+]i-wave propagation. This inhibition did not depend on the oxidizing effects of PMA because the addition of glaucine, a known antioxidant, did not prevent the inhibition. Cells grown for 72 hours in glucose-rich medium (25 or 50 mM) and in which all protein kinase C activity was downregulated by a previous 72-hour exposure to 1 microM PMA, did not display the inhibitory effect on the propagation of the Ca2+ wave that is normally induced by this elevated glucose level. Stimulation or inhibition of protein kinase A activity by incubating RPE cells with Sp-cyclic adenosine monophosphate or Rp-cyclic adenosine monophosphate respectively, or inhibition of tyrosine kinase activity with herbimycin A did not alter the intercellular communication after mechanical stimulation. To determine whether the observed changes were caused by alterations in gap junction conductance (GJC), FRAP experiments were performed in control conditions, after a 30-minute incubation with PMA, and in cells cultured in 50 mM glucose in the presence and in the absence of 1 microM PMA. The measured GJC was consistent with the inhibitory effect on propagation of an intercellular Ca2+ wave in all tested conditions. CONCLUSIONS: In RPE cells, a glucose concentration of 14 mM (224 mg/dl) or higher inhibits Ca(2+)-wave propagation and intercellular GJC. This effect may be mediated by protein kinase C activity. PMID- 9224289 TI - Role of P-selectin in endotoxin-induced uveitis. AB - PURPOSE: P-selectin is one of the early-reactive adhesion molecules that play a part in the rolling phase of leukocytes in cellular infiltration. The study objective was to determine whether P-selectin is involved in the development of endotoxin-induced uveitis (EIU). METHODS: Endotoxin-induced uveitis was initiated in male Lewis rats by injecting 200 micrograms lipopolysaccharide (LPS) into the foot pad. Expression of P-selectin in the iris-ciliary body was studied by immunohistochemistry using wholemounts and paraffin embedded sections of iris ciliary body prepared at various time intervals. A monoclonal antibody (mAb) against P-selectin and a ligand for P-selectin, sialyl Lewis X oligosaccharide (SLeX-OS), was intravenously injected to evaluate the effects of selectin inhibition. The effect of treatment was evaluated by the number of infiltrated cells and protein concentration in the aqueous humor at 24 hours after LPS treatment. RESULTS: P-selectin immunoreactivities were observed in the vessels of the iris in whole iris-ciliary body mounts and on the surface of the microvascular endothelium in paraffin-embedded sections. Activity was most prominent at 15 minutes and at 5 to 7 hours after LPS treatment and was moderate from 1 to 4 hours after treatment. The selective inhibition of P-selectin significantly blocked the cellular infiltration into aqueous humor, but this infiltration was even more effectively inhibited by SLeX-OS. Protein concentration in the aqueous humor was not inhibited by selectin as much as was cellular infiltration. CONCLUSIONS: In the early phase of EIU, P-selectin may be expressed on the vascular endothelium in the iris in a biphasic pattern that modulates the rolling phase of leukocytes. The expression of this molecule may be essential for succeeding processes of cellular infiltration and may determine the subsequent states of ocular inflammation. PMID- 9224290 TI - Immunity and immune privilege elicited by cultured retinal pigment epithelial cell transplants. AB - PURPOSE: To determine whether cultured retinal pigment epithelial (RPE) cells implanted in the subconjunctival space induce an immune response against autoantigens and whether an active downregulation is achieved by RPE grafts placed in the anterior chamber and within the subretinal space. METHODS: Cultured RPE cells from eyes of newborn C57BL/6 mice were implanted in the subconjunctival space, the anterior chamber, or the subretinal space of eyes of adult C57BL/6 mice. At postimplantation day 12, the recipients were evaluated for RPE-specific delayed hypersensitivity and examined clinically and histologically for evidence of rejection. To facilitate their identification, RPE cells were labeled with 5 bromodeoxyuridine, before intraocular transplantation. RESULTS: Cultured RPE cells implanted in the subconjunctival space of syngeneic mice elicited an intense RPE-specific delayed hypersensitivity associated with a vehement cellular infiltration of the graft when examined at postimplantation day 12. By contrast, grafts in the anterior chamber and subretinal space displayed no evidence of rejection, and their recipients failed to display RPE-specific delayed hypersensitivity. Additionally, the spleens of these mice contained regulatory T cells that suppressed RPE-specific delayed hypersensitivity in naive syngeneic recipients. CONCLUSIONS: Cultured RPE cells can induce an immune response against autoantigens. Implantation of RPE cells in immune-privileged sites of the eye induces a deviant immune response that is associated with spleen cells that suppress RPE-specific delayed hypersensitivity and autoimmune rejection. PMID- 9224291 TI - Immunocytochemical study of dystrophin localization in cone cells of mouse retinas. AB - PURPOSE: Previously, the authors reported that dystrophin was observed under the rod cell membranes in rat retinas. However, it was not determined whether dystrophin is located in cone cells. In the current study, the authors clarify dystrophin localization in cone cells of mouse retinas. METHODS: Immunoblotting, confocal laser scanning microscopy, and immunoelectron microscopy were used to investigate retinal dystrophin with a monoclonal antibody raised against the human dystrophin C-terminus. RESULTS: Immunoblotting analysis showed some immunoreactive bands from retinal extracts. Confocal images indicated two different immunostaining patterns: One was a tiny dot, and the other was a larger, aggregated dot. Immunoelectron microscopy revealed that retinal dystrophin was localized in cone cells as well as in rod cells. CONCLUSIONS: Retinal dystrophin is a common component of cone and rod cells and probably is related to the physiological function of photoreceptor cells. PMID- 9224292 TI - Treatment of bleb leaks with transforming growth factor-beta in the rabbit model. AB - PURPOSE: The mechanism through which peribleb injection of autologous blood results in resolution of bleb leak in the rabbit model remains unclear. This study evaluates the clinical and histologic effects of peribleb injection of transforming growth factor-beta (TGF-beta) after leak induction in mitomycin-C treated blebs. METHOD: Posterior lip sclerectomies treated with mitomycin-C were created in New Zealand White rabbits. On postoperative day 7, a standardized stab incision was performed on all blebs, and the eyes were randomized to receive a peribleb injection of either TGF-beta or of a balanced salt solution. RESULTS: Injection of TGF-beta was associated with the resolution of bleb leak and maintenance of a functioning bleb in 50% (4 of 8) of treated eyes. The remaining TGF-beta-treated eyes and control eyes demonstrated continued bleb leaks or bleb failures with intraocular pressure returning to preoperative levels. Histologic examination revealed increased peribleb cellularity and denser collagen deposition in the TGF-beta-treated eyes compared with that observed in control eyes. CONCLUSIONS: Peribleb TGF-beta injections may contribute to healing bleb leaks, but the injections do not appear in this model to be as useful as whole blood injections. PMID- 9224293 TI - Nitric oxide enhancement of cholinergic amacrine activity by inhibition of glycine release. AB - PURPOSE: To investigate a possible interaction between cholinergic and nitrergic amacrine cells in the rabbit retina. METHODS: The activity of cholinergic amacrine cells was estimated by measuring the light-evoked release of [3H] acetylcholine (ACh) from the retina of rabbits anesthetized with urethane. An eyecup was prepared and filled with Krebs-Ringer bicarbonate solution, containing [3H]-choline. After washing with fresh medium containing physostigmine, 0.5 ml of medium was placed in the eyecup. The medium was replaced every 5 minutes, and the radioactivity in the resultant samples was measured. In some experiments the release of [3H]-ACh and glycine was measured using isolated retinas. RESULTS: Local application of the nitric oxide (NO) donors, S-nitroso-N-acetyl-DL penicillamine and sodium nitroprusside strikingly enhanced the light-evoked release of [3H]-ACh. In contrast, inhibition of nitric oxide synthase with L nitromonomethylarginine (LNMMA) or N-nitro-L-arginine (LNA) greatly reduced the light-evoked release of [3H]-ACh. In that the response of cholinergic amacrine cells is damped by an inhibitory feedback circuit involving glycinergic amacrine cells, the effect of strychnine on the inhibitory action of LNMMA was examined, Strychnine abolished the inhibitory effect of LNMMA on the light-evoked release of [3H]-ACh, suggesting that endogenous NO normally has an inhibitory effect on glycinergic amacrine cells. This idea was supported by experiments using isolated retinas, in which sodium nitroprusside and S-nitroso-N-acetyl-DL-penicillamine inhibited the potassium-evoked release of glycine but enhanced the release of [3H]-ACh. CONCLUSIONS: Endogenous NO is released in the retina and acts indirectly to facilitate the light-evoked response of cholinergic amacrine cells. PMID- 9224295 TI - This month in investigative urology. Progressive changes in detrusor function with bladder outlet obstruction. PMID- 9224294 TI - Effect of varying the mitomycin-C treatment area in glaucoma filtration surgery in the rabbit. AB - PURPOSE: To investigate the effect of varying the treatment area of subconjunctival mitomycin-C (MMC) using an adapted rabbit model of filtration surgery. METHODS: Twenty-four New Zealand White rabbits underwent filtration surgery, with random allocation to one of three treatments: 5-minute subconjunctival applications of MMC (0.4 mg/ml) with either a large (8 x 10 mm) or small (4 x 2 mm) sponge or no treatment (control). Drainage was achieved by placing an intravenous cannula through a scleral tunnel into the anterior chamber. Rabbits were examined at set intervals for up to 30 days after surgery. Measurements of appearance, size, height, and vascularity of blebs and of intraocular pressure and anterior chamber depth were made by a masked observer. Histologic analysis of eyes was performed at 3, 14, and 30 days. RESULTS: Statistical analysis showed a significant difference in bleb survival among all groups (log rank P = 0.0054, with 100% survival with large areas of MMC treatment). Comparison between large and small treatment area groups revealed significant differences in bleb survival (log rank P = 0.0388); bleb area (between-subject analysis, P = 0.009), and bleb height (between-subject analysis, P = 0.005). These differences were seen clinically, with large areas of MMC treatment producing diffuse and elevated blebs, small areas of treatment producing thin-walled and localized blebs with scarring at 21 days, and no treatment resulting in comparatively vascularized and scarred blebs before 14 days. Histologic analysis revealed clear differences among groups, with an increase in subconjunctival cellularity and scar tissue in eyes with failed blebs. CONCLUSIONS: The size of the area of subconjunctival MMC treatment significantly affects surgical outcome. Histologic features mirror differences observed clinically. Alteration of the size of the MMC treatment area may provide an alternative and more controllable approach to modulating the wound-healing response after drainage surgery and, more important in the clinical context, to modifying bleb morphology. PMID- 9224296 TI - Treatment options for localized prostate cancer based on pretreatment serum prostate specific antigen levels. AB - PURPOSE: We reviewed all available literature on early stage prostate cancer treatment in which pretreatment serum prostate specific antigen (PSA) levels were used to stratify patients. We determined if any conclusions could be reached regarding the optimal therapy of this disease. MATERIALS AND METHODS: A MEDLINE search was conducted to obtain all articles in English on prostate cancer treatment from 1986 to 1996 in which PSA levels were used to stratify patients and evaluate outcome. Studies were considered eligible only if they met all criteria of pretreatment PSA values recorded and grouped for subsequent evaluation, posttreatment PSA values monitored continuously, definitions of biochemical control stated and median followup given. RESULTS: Of the 16 surgical studies identified only 3 met the inclusion criteria. Of the 30 radiation therapy articles identified 15 met the inclusion criteria, including 2 on conformal external beam radiotherapy, 8 on conventional external beam radiotherapy and 5 on interstitial brachytherapy. No studies using neutrons or combined hormonal therapy with surgery or radiotherapy were identified in which patients were stratified by pretreatment PSA. Results for all therapies were extremely variable with the 3 to 5-year rates of biochemical control ranging from 48 to 100% for patients with a pretreatment PSA of less than 4 ng./ml., 44 to 90% for PSA more than 4 to 10 ng./ml. and 27 to 89% for PSA more than 10 to 20 ng./ml. Even using the same treatment modality, a wide range of results were obtained. No treatment option consistently produced superior results. CONCLUSIONS: When data were reviewed from studies using pretreatment serum PSA to stratify patients, no consistently superior treatment option in the radiotherapy or surgical literature emerged. These data suggest that standard definitions of disease stage and biochemical cure must be adopted to evaluate treatment efficacy and advise patients on the most appropriate treatment option for the disease. PMID- 9224297 TI - Reverse transcriptase polymerase chain reaction for prostate specific antigen in the management of prostate cancer. AB - PURPOSE: Polymerase chain reaction is a powerful tool for expanding minute quantities of deoxyribonucleic acid (DNA) for detailed study. Reverse transcriptase polymerase chain reaction (RT-PCR) involves the initial conversion of messenger ribonucleic acid (mRNA) to DNA, followed by the amplification of the DNA product for molecular analysis. The mRNA for prostate specific antigen (PSA) is expressed only by prostatic cells. RT-PCR offers a potentially more sensitive assay for the detection of cells expressing PSA mRNA in peripheral circulation or in extraprostatic tissues. The current status of RT-PCR in the amplification and detection of PSA gene expression in the management of prostate cancer is reviewed. MATERIALS AND METHODS: The literature was reviewed for available data using RT-PCR for detection of prostatic cells outside of the prostate. RESULTS: Amplification of mRNA by RT-PCR represents a highly sensitive method of detection of gene expression. A single cell expressing PSA among 10 to 100 million lymphocytes can be detected by the RT-PCR assay. This assay may detect extraprostatic or circulating prostatic cells in peripheral blood, lymph nodes and bone marrow in many patients with prostate cancer. Various studies have reported sensitivities of detection of PSA expressing cells in the peripheral blood ranging from 0 to 88%. This wide range in the sensitivity may be partly due to tremendous variation between the protocols used in each study. In patients with lymph node metastasis the RT-PCR assay appears more reliable than immunohistochemistry for identification of prostatic tissue in the lymph node. In some series analyses of radical prostatectomy specimens suggest a strong correlation between a positive RT-PCR result and capsular invasion by the tumor, while others do not support its use in determining pathological stage. In the majority of reports the RT-PCR assay was highly specific for detection of extraprostatic PSA expression in prostate cancer patients, and negative for detection in men with benign prostatic hyperplasia and in women. Sources of potential false-positive and false-negative results of this assay are identified and discussed. CONCLUSIONS: RT-PCR can detect PSA expressing cells that are otherwise undetectable by other means in patients with localized and metastatic cancers. This assay is highly specific, since PSA expressing cells were consistently undetectable in the peripheral circulation of patients without prostate cancer in most studies. Limited data to date suggest that this test may have a role in the staging of tumors before radical prostatectomy and in the serial followup of patients after treatment. RT-PCR may improve the detectability of lymph node metastasis over immunohistochemistry. Presently this test should remain a powerful research tool in the study of the biology and behavior of prostate cancer, and it should not be used to guide any clinical decision making. The use of this assay as a prognostic and management tool for prostate cancer is in the earliest stages. PMID- 9224298 TI - Initial evaluation of CYFRA 21-1 diagnostic performances as a urinary marker in bladder transitional cell carcinoma. AB - PURPOSE: CYFRA 21-1, an immunoradiometric assay developed for the detection of a soluble cytokeratin 19 fragment, is evaluated for its diagnostic performance in urine of patients with transitional cell carcinoma. MATERIALS AND METHODS: CYFRA 21-1 was investigated in serum and urine of 128 patients, including 48 with bladder transitional cell carcinoma (group 1), 44 with other urological pathological conditions (group 2) and 36 free of urothelial disease (group 3). Urinary cytopathology was also performed. RESULTS: Mean urinary CYFRA was 123.5 +/- 53, 11.9 +/- 4.8 and 2.3 +/- 0.2 ng./ml. for groups 1 to 3, respectively, and was significantly different. From the receiver operating characteristics curve, the optimal combination of 96% sensitivity and 74% specificity was determined for a threshold value of 4 ng./ml. while overall cytopathology sensitivity was 43%. CONCLUSIONS: Urinary CYFRA 21-1 may be a useful marker for diagnosing transitional cell carcinoma. PMID- 9224299 TI - Combination "sandwich" therapy for extensive renal calculi in 100 consecutive patients: immediate, long-term and stratified results from a 10-year experience. AB - PURPOSE: We determined the immediate and long-term efficacy of combination "sandwich" therapy for management of large, extensively branched calculi in 100 consecutively treated patients. MATERIALS AND METHODS: We treated 61 women and 39 men for stones ranging from 2.2 to 66 cm2 (mean 20.8) with percutaneous debulking followed by shock wave lithotripsy and, when necessary, secondary nephroscopy via the mature tract. The primary debulking was performed via 1 to 3 tracts (total 106, mean 1.06 per patient), following which 1 to 3 shock wave treatments (total 127, mean 1.3 per patient) were administered. Subsequently, 62 patients underwent 71 secondary or tertiary percutaneous procedures (mean 1.1 per patient). RESULTS: Total hospital stay ranged from 3 to 44 nights (mean 12.2) and decreased with experience. In 34 patients 40 complications developed, the most frequent of which were bleeding requiring transfusion in 14 patients and fever or sepsis delaying a planned procedure or hospital discharge in 20 patients. For patients with struvite stones the transfusion rate and fever/sepsis rate was 20 and 33%, respectively, compared to only 10 and 12%, respectively, for those patients with noninfection related stones. Of 87 patients available for 1-month radiographic followup 55 (63%) were stone-free, while 32 (37%) had discrete residual gravel. With time and experience, the stone-free rate improved from 52 to 70%. Of 55 patients followed for a mean of 40.5 months ipsilateral stones recurred in 13 (22.8%). Of 39 patients with struvite calculi 11 (28%) had recurrent bacteriuria or infection. Renal function, defined by serum creatinine, ranged from 0.6 to 3.9 mg./dl. (mean 1.3) before treatment and from 0.5 to 6.4 mg./dl. (mean 1.4) 1 to 101 months (mean 31) after treatment. CONCLUSIONS: This combined sandwich approach offers immediate and long-term results comparable to other forms of management currently available for these challenging cases. Furthermore, this approach may be applied successfully to virtually any patient with large, extensively branched or otherwise complex stones. PMID- 9224300 TI - New onset hypertension after extracorporeal shock wave lithotripsy: age related incidence and prediction by intrarenal resistive index. AB - PURPOSE: In a recent study we found an increased resistive index immediately after extracorporeal shock wave lithotripsy (ESWL) in patients older than 60 years, which suggests renovascular disturbance. The present 26-month followup study was undertaken to investigate the relevance of elevated resistive index levels and the incidence of new onset hypertension. MATERIALS AND METHODS: Of the initial 76 patients 57, including 20 of the 23 at risk patients 60 or greater years, group 3), were followed for more than 26 +/- 6 months after ESWL. Followup included 2 resistive index measurements by Doppler ultrasound of the treated and the contralateral kidney, at least 2 blood pressure measurements 1 week apart and excretory urography as well as determination of plasma renin activity in 9 patients. RESULTS: With 1 exception, elevated resistive index levels and hypertension were observed exclusively in patients older than 60 years. In these patients the resistive index ranged between 0.65 and 0.86 (mean plus or minus standard deviation 0.74 +/- 0.05, normal less than 0.7). This increase in resistive index was statistically significant (p < 0.0001). Compared to the levels obtained immediately after ESWL, the resistive index continued to increase in all 9 patients older than 60 years who had hypertension (45%), whereas in the normotensive patients the resistive index was either stable or decreased. There was a strong positive correlation (0.903) between pathological resistive index levels and blood pressure. CONCLUSIONS: Patients older than 60 years are at risk for disturbances of renal perfusion as assessed by the resistive index, and 45% of these patients have new onset hypertension within 26 months of treatment. PMID- 9224302 TI - Leave no stone unturned? PMID- 9224301 TI - Renal stone fragments following shock wave lithotripsy. AB - PURPOSE: We describe a select group of asymptomatic patients with fragments and dust 3 months after extracorporeal treatment, who were followed to evaluate the long-term outcome and therapeutic implications. MATERIALS AND METHODS: A total of 129 patients with dust and residual fragments (less than 4 mm.) at 3 months was re-examined at 12 months, and 95 were also evaluated at 24 months. Followup examinations consisted of radiographic studies, renal ultrasonography and urine culture. Dust and residual fragments were sought, and patients were defined as free or as having persistent lithiasis or stone regrowth. At 24 months recurrences in the patients stone-free at 12 months also were considered. RESULTS: At the 12-month followup 60 patients (46.5%) were stone-free and 56 (43.5%) still had dust or residual fragments. The localization of the stones or fragments at 3 months and their sizes did not have a significant influence on the stone-free rate but regrowth was greater in patients with stones larger than 10 mm. (11 of 40 patients, 27.5% versus 2 of 89, 2.2%, p = 0.001). The probability of eliminating residual lithiasis at 12 months was significantly greater in patients with dust than in those with residual fragments (42 of 79 patients, 58% versus 18 of 50, 36%, p = 0.026). Regrowth of residual lithiasis was observed in 13 patients (10%). CONCLUSIONS: Based on our results, we do not believe that patients with fragments require systematic re-treatment in the short term but they may be followed long term and re-treated if symptoms persist or stones recur. PMID- 9224303 TI - Experience with revascularizing renal artery aneurysms: is it feasible, safe and worth attempting? AB - PURPOSE: We retrospectively evaluated the feasibility and efficacy of surgical revascularization for renal artery aneurysms. MATERIALS AND METHODS: Beginning in 1984, 12 patients with renal artery aneurysm underwent renal revascularization regardless of clinical features. Postoperative results were analyzed regarding split renal function, patency of the revascularized arteries, blood pressure control and surgical complications. RESULTS: Postoperative renal function was stable or improved in all but 1 case and patency in branched arteries was preserved in 86%. Hypertension in 8 patients was cured in 7 (88%), including 2 with renovascular hypertension, and improved in 1 (12%). Complications were minimal with only 1 ureteral stricture that required reoperation. CONCLUSIONS: The majority of renal artery aneurysm cases are amenable to surgical repair. Carefully performed renal revascularization is rewarding in that high blood pressure is better controlled, renal function is improved and the potential risk of rupture is obviated. PMID- 9224304 TI - Incisional hernia after laparoscopic nephrectomy with intact specimen removal: caveat emptor. AB - PURPOSE: We report 5 cases of postoperative incisional hernia after laparoscopic nephrectomy with intact removal of the specimen. MATERIALS AND METHODS: During the last 5 years 29 patients underwent laparoscopic nephrectomy with intact removal of the resected specimen due to a large kidney and/or malignancy. Of these 29 patients 5 had a postoperative incisional hernia at the site of intact removal, including 3 with renal tumors and 2 with large polycystic kidneys due to adult onset autosomal dominant polycystic kidney disease. The records of these patients were reviewed to determine any specific factors that might relate to the development of this complication. RESULTS: An incisional hernia developed at the wound site in 5 patients (17%) 41 to 73 years old (mean age 53.4). Average body mass index for the patients was 34.2 (range 26 to 47). Average weight and size were 542 gm. and 20.3 x 10.3 cm., respectively, for the 3 resected malignant specimens and 1,975 gm. and 23.8 x 16.5 cm., respectively, for the 2 benign kidneys. A transverse lower flank muscle cutting incision (average 10.4 cm.) was performed to remove the resected kidney. Incisional hernias appeared after an average of 6.6 weeks postoperatively. Risk factors for a postoperative hernia included obesity in 80% of the patients, chronic renal insufficiency due to autosomal dominant polycystic kidney disease in 40%, postoperative pulmonary complication in 40% and metastatic cancer in 20%. CONCLUSIONS: Our experience has led us to avoid a lower flank port connecting incision for specimen removal. Instead we changed to a midline or subcostal incision in these patients. In addition, we believe that with the availability of the impermeable organ entrapment sacks there is less need for intact specimen removal even for renal tumors. Currently large benign kidneys (autosomal dominant polycystic kidney disease) are morcellated in situ to a suitable size for entrapment, while renal tumors are entrapped and morcellated directly. Presently our only indication for intact removal is in the case of a renal pelvic or caliceal transitional cell cancer. PMID- 9224305 TI - Simultaneous chromosome 7 and 17 gain and sex chromosome loss provide evidence that renal metanephric adenoma is related to papillary renal cell carcinoma. AB - PURPOSE: Metanephric adenoma has recently been recognized as a unique renal tumor characterized by an unusual degree of cellular differentiation and maturation. We recently studied metanephric adenoma using metaphase analysis and observed concomitant chromosome Y loss and chromosome 7 and 17 gain. To determine if these chromosomal anomalies are consistently present in renal metanephric adenoma, we studied all 11 tumors in the pathology tissue registry at our institution using fluorescence in situ hybridization (FISH). MATERIALS AND METHODS: FISH, using deoxyribonucleic acid probes for chromosomes 1, 7, 8, 17, X and Y, was performed in isolated nuclei from 11 paraffin embedded renal metanephric adenoma specimens. RESULTS: Of the 11 tumors (73%) 8 demonstrated chromosome 7 and 17 gain by FISH, and the remaining 3 were found to have an apparently normal chromosomal content. Of the 8 tumors (75%) from men showed 6 chromosome 7 and 17 gain with Y chromosome loss. Of the 3 tumors (33%) from women 1 had chromosome 7 and 17 gain with X chromosome loss, while 1 had chromosome 7 and 17 gain without sex chromosome aneusomy. Metaphase analysis performed on 2 tumors revealed chromosome 7 and 17 gain and Y chromosome loss in 1, and no apparent, chromosome anomaly in the other, confirming the results of FISH analysis. CONCLUSIONS: FISH analysis of renal metanephric adenoma identified frequent chromosome 7 and 17 gain and sex chromosome loss. These results are consistent with a clonal neoplastic disorder in which chromosomes 7, 17, X and Y are likely to be involved in the pathogenesis of this tumor. These characteristic chromosomal alterations have also been observed in papillary renal cell adenoma and papillary renal cell carcinoma, providing evidence that these tumors may be related. PMID- 9224306 TI - Surgery for metachronous solitary liver metastases of renal cell carcinoma. AB - PURPOSE: The postoperative outcome and survival of patients undergoing surgery for metachronous solitary liver metastases of renal cell carcinoma were evaluated. MATERIALS AND METHODS: Between 1983 and 1993, 17 patients with metachronous liver metastases of renal cell carcinoma underwent laparotomy for metastatic liver disease. All patients had undergone radical nephrectomy a mean of 3.6 years before the diagnosis of liver metastases. RESULTS: Surgical resection was feasible in 13 of 17 patients with right hemihepatectomy in 9 (3 multivisceral resections), wedge resection in 4 and ex situ (mobilization and eversion out of the abdomen) resection in 1. Stage R0 resection (complete removal, negative surgical margins with no macroscopic disease left behind) was possible in 11 of 13 cases (85%). In patients with metastatic liver tissue resection the mortality rate was 31% (4 of 13) with additional significant morbidity in another 2. Mean survival of patients with nonresectable disease was 4 months, which increased to 16 months after resection. CONCLUSIONS: Complete resection of metachronous liver metastases can be achieved in the majority of patients. However, significant morbidity and mortality as well as the limited prognosis even after R0 resection strongly suggest careful patient selection. PMID- 9224308 TI - Prognostic value of Ki-67 for recurrence and progression of superficial bladder cancer. AB - PURPOSE: We retrospectively reviewed 104 cases of superficial bladder cancer to ascertain whether the Ki-67 labeling index predicts recurrence and progression. MATERIALS AND METHODS: Archival specimens from superficial bladder cancer cases were immunostained with Ki-67. RESULTS: The recurrence rate was significantly higher in cases with a Ki-67 labeling index of 5.35 or greater than in those with a value less than 5.35 (p < 0.001). The recurrence rate at 2 years was 70% in cases with a Ki-67 labeling index of 5.35 or greater and 22% in those with an index less than 5.35 (p < 0.001). Using multivariate analysis the index predicted recurrence of bladder cancer (p < 0.005). Median Ki-67 labeling index in cases with progression was significantly higher than in those without progression (p < 0.01). CONCLUSIONS: The Ki-67 labeling index is an independent predictive factor for recurrence of superficial bladder cancer. PMID- 9224307 TI - Adjuvant chemotherapy for superficial transitional cell bladder carcinoma: long term results of a European Organization for Research and Treatment of Cancer randomized trial comparing doxorubicin, ethoglucid and transurethral resection alone. AB - PURPOSE: We compared the efficacy of transurethral resection alone or transurethral resection followed by bladder instillations of doxorubicin or ethoglucid for 1 year in patients with superficial bladder carcinoma, and followed them long term for the incidence of progression to muscle invasion. MATERIALS AND METHODS: A total of 443 patients with superficial transitional cell carcinoma of the bladder was randomized. After randomization of 206 patients the control arm was closed to patient entry based on the results of an interim analysis showing a significant difference in favor of those receiving adjuvant chemotherapy. RESULTS: Final analysis of treatment results for recurrence included 432 patients at a median followup of 3.4 years for time to first recurrence, 5 years for analysis of time to invasion (Category T2 disease or worse) and 10.7 years for duration of survival. Time to first recurrence was significantly prolonged by both drugs compared to transurethral resection alone (doxorubicin versus transurethral resection alone p < 0.001 and ethoglucid versus control p < 0.001). Recurrence rate per year was 0.30 for both adjuvant treatment arms and 0.68 for the resection only group. Progression to muscle invasion was rare (15.1% of cases) and not apparently different in the 3 treatment arms. Of the 423 patients death from any cause in 199 and from malignant disease in 59 was not correlated with treatment. However, there was a strong correlation between death from malignant disease, and T category and tumor grade. CONCLUSIONS: In regard to time to first recurrence and recurrence rate per year this study indicates that adjuvant chemotherapy with doxorubicin and ethoglucid using the indicated schedule is superior to transurethral resection alone. However, progression in stage or survival was not influenced by the treatment regimen. PMID- 9224309 TI - Long-term followup of all patients with muscle invasive (stages T2, T3 and T4) bladder carcinoma in a geographical region. AB - PURPOSE: We studied the relationship between long-term survival and treatment of stages T2, T3 and T4 bladder carcinoma in an unselected patient population. MATERIALS AND METHODS: A total of 680 patients with the initial diagnosis of bladder carcinoma in 1987 to 1988 in Western Sweden was prospectively registered and followed until 1994. Of these patients 107 had stage T2 to T3 and 41 had stage T4 disease. RESULTS: Of the patients with stage T2 to T3 disease 30 (mean age 66) underwent radical cystectomy, 33 (mean age 75) full dose radiotherapy and 44 (mean age 81) nonradical therapy (mainly transurethral resection of the bladder). The 5-year crude survival rates were 33, 15 and 14%, respectively. Of the patients with stage T4 disease 6 (mean age 61) underwent radical cystectomy, 9 (mean age 73) full dose radiotherapy and 26 (mean age 81) nonradical therapy (mainly transurethral resection of the bladder). All except 1 patient died of disease within 4 years. CONCLUSIONS: More than 60% of the patients in the cohort were considered unsuitable for radical cystectomy and their survival was poor, whether treated with full dose radiotherapy or transurethral resection of the bladder alone. PMID- 9224310 TI - Radical cystectomy for carcinoma of the bladder: critical evaluation of the results in 1,026 cases. AB - PURPOSE: We performed a critical analysis of the different prognostic factors affecting survival among patients with carcinoma of the bladder for whom cystectomy was indicated. The different patient and tumor characteristics were correlated to survival data by a univariate as well as multivariate analysis. MATERIALS AND METHODS: Between 1969 and 1990, 764 men and 262 women, average age plus or minus standard deviation 43 +/- 8 years, with invasive carcinoma of the bladder were eligible for 1-stage radical cystectomy and urinary diversion. Patients were followed regularly and examined signs for and location of treatment failure. Followup ranged from 0 to 24.2 years, with a median plus or minus standard deviation of 4.05 +/- 4.16 years. RESULTS: Postoperative mortality was 4%. Most of the patients presented with advanced stage (greater than P3) disease. Squamous tumors accounted for 59% of cases, transitional carcinoma 22% and adenocarcinoma 11%. Bilharzial eggs were seen in 85% of the specimens. Regional lymph nodes were involved in 18.3% of the cases. The 5-year survival rate was 48%. The survival estimate was correlated to patient and tumor characteristics by univariate and multivariate analyses. Only tumor stage and grade, and lymph node status had a significant impact on survival. CONCLUSIONS: Contemporary cystectomy with continent diversion for muscle invasive disease provides minimal morbidity, offers good locoregional disease control and results in acceptable quality of life. The presence of positive regional lymph nodes is not a contraindication to this therapy. PMID- 9224311 TI - Orthotopic lower urinary tract reconstruction in women using the Kock ileal neobladder: updated experience in 34 patients. AB - PURPOSE: Orthotopic lower urinary tract reconstruction has revolutionized urinary diversion following cystectomy. Initially performed solely in male patients, orthotopic diversion has now become a viable option in women. Currently, the orthotopic neobladder is the diversion of choice for women requiring lower urinary tract reconstruction at our institution. We evaluate and update our clinical and functional experience with orthotopic reconstruction in female patients. MATERIALS AND METHODS: Since June 1990, 34 women 31 to 86 years old (median age 67) have undergone orthotopic lower urinary tract reconstruction following cystectomy. Indications for cystectomy included transitional cell carcinoma in 29 patients, urachal adenocarcinoma in 1, mesenchymal tumor of endometrial origin in 1, cervical carcinoma in 1 and a fibrotic radiated bladder in 1. In addition, 1 woman underwent undiversion to the native urethra following a previous simple cystectomy and cutaneous diversion for eosinophilic cystitis. Data were analyzed according to postoperative early and late complications, survival, tumor recurrence, pathological evaluation of the cystectomy specimen, continence status, voiding pattern and patient satisfaction. The median followup in this group of patients was 30 months (range 17 to 70). RESULTS: There were no perioperative deaths, and 4 early (11%) and 3 (9%) late complications. Four patients died, none with a urethral recurrence, including 3 of metastatic bladder cancer and 1 of unrelated causes. In another patient with an extensive mesenchymal tumor of the uterus a sigmoid tumor recurred requiring conversion of the orthotopic reservoir to a cutaneous diversion. All of the remaining 29 patients are alive without evidence of disease. Intraoperative frozen section of the distal surgical margin (proximal urethra) accurately evaluated (confirmed by permanent section) the proximal urethra prospectively for tumor in all 29 specimens removed for transitional cell carcinoma, including 28 specimens (97%) without evidence of tumor and 1 specimen with carcinoma in situ. Complete daytime and nighttime continence was reported by 29 (88%) and 27 (82%) of 33 evaluable patients, respectively. A total of 28 patients (85%) void to completion, while 5 (15%) require some form of intermittent catheterization to empty the neobladder. Patient satisfaction is overwhelming. CONCLUSIONS: The excellent clinical and functional results demonstrated with further followup confirm our initial experience with orthotopic diversion in women. Careful selection of appropriate female candidates for orthotopic diversion is critical, and includes preoperative evaluation of the bladder neck and intraoperative frozen section analysis of the distal cystectomy margin. Furthermore, close monitoring of the retained urethra is mandatory in all women undergoing orthotopic diversion. We believe that the orthotopic neobladder is the urinary diversion of choice in women following cystectomy. PMID- 9224312 TI - A worldwide view of bladder cancer. PMID- 9224313 TI - Cisplatin, ifosfamide, methotrexate and vinblastine combination chemotherapy for metastatic urothelial cancer. AB - PURPOSE: We investigated the activity of combination chemotherapy consisting of cisplatin, ifosfamide, methotrexate and vinblastine in patients with metastatic urothelial cancer. MATERIALS AND METHODS: A total of 32 consecutive patients was treated with 30 mg./m.2 cisplatin on days 1 through 3, 1.5 gm./m.2 ifosfamide with mesna on days 1 through 3, 30 mg./m.2 methotrexate and 3 mg./m.2 vinblastine on day 1 plus 5 micrograms./kg. granulocyte colonystimulating factor on days 7 through 11. Courses were repeated every 21 days for a maximum of 6 cycles. RESULTS: Major toxicity was granulocytopenia in 56% of patients, including 11 episodes of granulocytopenic fever. Anemia and thrombocytopenia developed in a third of the cases. No other significant toxicity or treatment related death was noted. An objective response was achieved in 20 patients (62.5%, 95% confidence interval 44 to 79). Median time to progression was 7 months and median survival was 13 months. CONCLUSIONS: The cisplatin, ifosfamide, methotrexate and vinblastine regimen appeared active in patients with metastatic urothelial carcinoma. This regimen was associated with significant but manageable hematological toxicity and the incidence of mucositis or renal impairment was low. Prospective randomized studies are needed to assess whether the addition of ifosfamide to other active agents will improve the survival of patients with this disease. PMID- 9224314 TI - A new biaxial epilated scrotal flap for reconstructive urethral surgery. AB - PURPOSE: We describe a new type of perineum based scrotal flap with biaxial vascularization supplied by both superficial perineal arteries. Flap length of up to 20 cm. may be attained for urethral reconstruction. MATERIALS AND METHODS: A total of 37 men with complex urethral stenosis of different etiologies underwent surgery using 1 of 3 urethroplasty techniques based on this new flap. The whole anterior urethra, including pendulous and bulbar segments, was reconstructed with a scrotal patch in 10 patients. A scrotal tubular flap was used as a substitute for the bulbar urethra in 7 patients and for the membranous portion in 4. Bulbar urethroplasty with a scrotal island patch was performed in 16 patients. RESULTS: Of the patients 86% achieved normal voiding after 1-stage urethroplasty. Mean followup was 39.5 months. CONCLUSIONS: The excellent axial vascularization of this new flap permits successful resolution of the most complex urethral stenoses regardless of extension, location and etiology. PMID- 9224315 TI - Experience with 30 posttraumatic rectourethral fistulas: presentation of posterior transsphincteric anterior rectal wall advancement. AB - PURPOSE: We present the challenging problems involving the treatment of rectourethral fistulas, especially those caused by war wounds. Various existing techniques used by a single surgeon are compared in this study. The method of posterior transsphincteric anterior rectal wall advancement is described as the treatment of choice. We emphasize the importance of fecal and urinary diversion. To our knowledge this series is the largest in the literature. MATERIALS AND METHODS: From 1981 to 1994 we treated 30 men 18 to 50 years old (mean age 34) with posttraumatic rectourethral fistulas, including 23 (76.5%) caused by missiles. Urethroscopy with digital examination under anesthesia was the most reliable diagnostic study. End sigmoid colostomy and suprapubic cystostomy were performed in all patients. RESULTS: In 14 patients (46.5%) the fistula healed after double diversion but 16 (53.5%) required reconstruction for repair. Of the 6 procedures using established techniques in 5 patients 3 (50%) failed and 3 were successful but a urethral stricture developed after 2 (66%). On the other hand, in all patients (100%) who underwent repair via posterior transsphincteric anterior rectal wall advancement the fistula resolved and a stricture developed in 3 (27%). Fistula size and extent of fibrosis affected treatment, while etiology did not. Urethral obstruction complicated only the missile wounds. CONCLUSIONS: Double diversion has resulted in resolution of approximately half of the small, less fibrous fistulas. Early repair is recommended for large fibrous fistulas. Anterior rectal wall advancement through a posterior transsphincteric incision offers a new option that has proved to be successful and safe, and causes fewer urethral complications. It also provided good visualization with minimal bleeding and was less painful. Double diversion is a prerequisite to reconstruction. PMID- 9224316 TI - Traumatic posterior urethral injury and early realignment using magnetic urethral catheters. AB - PURPOSE: We determined the success of early urethral realignment using magnetic urethral catheters. MATERIALS AND METHODS: We retrospectively reviewed the records of 13 patients with complete urethral disruption treated with endourological realignment 0 to 11 days after injury using coaxial magnetic urethral catheters. RESULTS: Urethral realignment was established in 11 of the 13 patients (85%) using magnetic urethral catheters. Of the 10 patients for whom followup was available urethral strictures developed in 5 (50%) a mean of 6.1 months after realignment, necessitating a mean of 1.4 corrective procedures per patient. Impotence was noted in 1 of 7 patients (14%) and no urinary incontinence developed after realignment. CONCLUSIONS: Urethral realignment within 2 weeks of injury using magnetic urethral catheters is a safe and simple technique with minimal morbidity. The stricture formation, impotence and incontinence rates of this technique are comparable to those reported for delayed urethroplasty. We advocate early realignment using magnetic urethral sounds as an alternative treatment for traumatic urethral disruption. PMID- 9224317 TI - Treatment results using pubovaginal slings in patients with large cystoceles and stress incontinence. AB - PURPOSE: We determined the efficacy of performing a pubovaginal sling concurrently with a formal cystocele repair in patients with grade III to IV cystoceles. MATERIALS AND METHODS: We studied 42 women with grade III to IV cystoceles diagnosed by physical examination and video urodynamics. Of the patients 9 (22%) had intrinsic sphincter deficiency diagnosed by an abdominal leak point pressure of less the 60 cm. water, and 24 (57%) had type II stress incontinence with urethral hypermobility and an abdominal leak point pressure greater than 90 cm. water. A pubovaginal sling and anterior colporrhaphy were performed and, if indicated, other vaginal procedures were done at that time. RESULTS: A total of 36 patients (86%) was available for postoperative pelvic examinations performed at 3-month intervals, for a mean followup of 20.4 months (range 12 to 39). Only 3 patients had symptomatic grade III cystoceles and 2 had enteroceles. Two patients required collagen injections and 2 underwent a repeat pubovaginal sling. Therefore, all patients were continent at the time of followup. CONCLUSIONS: This study confirms that in patients with large cystoceles and stress urinary incontinence a pubovaginal sling and anterior colporrhaphy effectively treat the incontinence and reduce the cystocele. In addition, the fascial sling appears to provide additional support to the bladder base, improving the durability of the anterior colporrhaphy. PMID- 9224318 TI - Quality of life and continence assessment of the artificial urinary sphincter in men with minimum 3.5 years of followup. AB - PURPOSE: We determined the long-term efficacy and quality of life impact of the artificial urinary sphincter. MATERIALS AND METHODS: We reviewed the medical records of 68 men who underwent artificial urinary sphincter placement for post prostatectomy incontinence (64) or neurogenic disease (4) between March 1980 and March 1992 (mean followup 7.2 years). Quality of life was assessed in 52 patients who completed the incontinence impact questionnaire and the urogenital distress inventory. RESULTS: At followup 54 men were socially continent (0 or 1 pad per day). Overall, pad score decreased significantly from 2.75 before to 0.97 after artificial urinary sphincter implantation (p < 0.001). The artificial urinary sphincter was permanently removed in 4 patients. Revisions for mechanical failure or urethral atrophy were required in 25% of the patients (mean 1.35 procedures per patient). The mechanical failure rate decreased significantly after 1987 due to device improvements (12.4 versus 44.4%, p < 0.01). Subjective improvement and overall satisfaction were rated as 4.1 and 3.9, respectively (scale 0 to 5). At followup the mean values of the incontinence impact questionnaire and urogenital distress inventory demonstrated the positive impact of the artificial urinary sphincter on quality of life. CONCLUSIONS: This long-term study documents the positive impact of the artificial urinary sphincter on patient quality of life with few mechanical failures since 1987. PMID- 9224319 TI - The evaluation of arterial inflow by gravity cavernosometry. AB - PURPOSE: We determined whether the comparison between equilibrium pressure after intracavernous injection of vasodilators and maximal corporeal pressure at gravity cavernosometry could provide information about the relative contribution of arterial inflow and cavernous wall resistance to the erection process. MATERIALS AND METHODS: The results of gravity cavernosometry performed in 68 impotent patients were compared to those of duplex scanning in 53 and penile angiography in 10. RESULTS: A highly statistically significant (p < 0.01) but nonlinear correlation was observed between the equilibrium pressure after injection and maximal corporeal pressure, which indicates a paramount role of the corporeal veno-occlusive mechanism in the development of penile rigidity. However, in most patients with a pressure increase of more than 30 mm. Hg from the equilibrium pressure after injection to the maximal corporeal pressure, arterial insufficiency was diagnosed by duplex scanning and/or arteriography, and seemed to be the main limiting factor in the development of penile rigidity. CONCLUSIONS: Gravity cavernosometry provides functional information about the corporeal veno-occlusive mechanism and arterial inflow and, therefore, about the relative roles of these mechanisms in the development of penile rigidity. PMID- 9224320 TI - Single potential analysis of corpus cavernosum electromyography for the assessment of erectile dysfunction: provocation, reproducibility and age dependence--findings in 36 healthy volunteers and 324 patients. AB - PURPOSE: Corpus cavernosum electromyography is a controversial method for assessing erectile failure. For its application as a diagnostic tool with clinical relevance, intra-individual stability of the parameters in independent recordings as well as information about provocation and age dependence are required. MATERIALS AND METHODS: We investigated reproducibility, provocation and age dependence of 11 parameters of single potential analysis of corpus cavernosum electromyography for 36 healthy volunteers in 2 independent recordings with surface electrodes using a visual evaluation technique. Recording 1 results were compared to findings for 324 men with erectile dysfunction. RESULTS: In healthy subjects all parameters varied extremely among individuals and they were poorly reproducible at repetition. The definition of a normal range as mean plus or minus 2.5 standard deviations did not result in useful diagnostic criteria for individual cases. Activity was not age dependent. Significant differences between healthy and impotent men as defined groups were found in provocation (overall maximum likelihood chi-square 15.5, dF = 2, p < 0.0005). CONCLUSIONS: Single potential analysis of corpus cavernosum electromyography seems to be appropriate for distinguishing potent volunteers from patients with erectile dysfunction. Provocation of slow cavernous electric activity seems to be a promising parameter that should be considered for ongoing studies. However, a high range of variation of findings even intra-individually does not currently qualify the method for routine clinical use. Further research will show whether different means of documentation or evaluation, that is corpus cavernosum electromyography pattern analysis at rest after audiovisual sexual stimulation, drug application or digital conversion of data, will lead to better results. PMID- 9224321 TI - Somatosensory evoked potentials in patients with primary premature ejaculation. AB - PURPOSE: Premature ejaculation has been believed to be psychological in the majority of patients. With few exceptions organic conditions are rarely implicated. We investigated the possible role of sensory function in patients with primary premature ejaculation to determine whether there is an etiological basis for this condition. MATERIALS AND METHODS: We performed somatosensory evoked potentials from the penis in 34 patients with primary premature ejaculation and in 30 normally potent men. The latencies and amplitudes of the evoked potentials were measured at the penile shaft (dorsal nerve) and at the glans penis. RESULTS: Mean latency of dorsal nerve and glans penis somatosensory evoked potentials was 1.51 and 6.80 (significant) msec. shorter, respectively, in the patients than in the normal subjects. In the normal subjects the mean latency of glans penis somatosensory evoked potentials was 0.99 msec. longer than that of the dorsal nerve (not significantly different) but in patients the mean latency in the glans penis was 4.30 msec. shorter (p < 0.001). Mean amplitude of glans penis somatosensory evoked potentials was less than that of the dorsal nerve in both groups. However, mean amplitudes of dorsal nerve and glans penis somatosensory evoked potentials were significantly greater in patients than in normal men. CONCLUSIONS: Patients with premature ejaculation have hypersensitivity and hyperexcitability of the glans penis, which may give rise to uncontrolled ejaculation and are believed to be organic implications for premature ejaculation. PMID- 9224322 TI - Long-term followup of and patient satisfaction with the Dynaflex self-contained inflatable penile prosthesis. AB - PURPOSE: We analyzed our experience with the Dynaflex* self-contained inflatable penile prosthesis to define specific complication rates and patient satisfaction in the long term. MATERIALS AND METHODS: From October 1990 through October 1994, 62 men underwent implantation of a Dynaflex prosthesis. Mean and median time since implantation was 50 and 53 months, respectively (minimum 24). In addition to standard followup and tabulation of complications, patients were contacted and interviewed regarding satisfaction with the prosthesis. RESULTS: Mechanical device failures occurred in 9.7% of cases with a mean time to failure of 40 months in 6 prostheses. An additional 16.1% of patients had an unsuccessful outcome due to dissatisfaction with the Dynaflex. Of the patients with a functional Dynaflex prosthesis contacted at the time of this review 88.1% expressed satisfaction. CONCLUSIONS: Mechanical failure rates for the Dynaflex prosthesis are comparable to those previously reported for multicomponent inflatable penile prostheses. Patient dissatisfaction, mainly due to difficulty in operating the Dynaflex inflation and deflation mechanisms, is much higher than for multicomponent inflatable penile prostheses. However, in patients who are successful in mastering operation of the Dynaflex satisfaction rates are high. Careful patient selection, and extended training and education efforts are the keys to maximizing successful outcomes with the Dynaflex prosthesis. PMID- 9224323 TI - Modified Nesbit procedure for the treatment of Peyronie's disease: a comparative outcome analysis. AB - PURPOSE: We describe a surgical modification of the Nesbit procedure to correct Peyronie's disease, and compare the results of this procedure with those of 2 other surgical techniques. MATERIALS AND METHODS: In 30 patients a vertical incision in the tunica albuginea was closed in a horizontal fashion with permanent suture knots buried beneath the tunica in a running looped fashion, resulting in watertight closure with no exposed suture material. A standard Nesbit procedure and plaque excision with a polyethylene terephthalate mesh reinforced silicone sheet patch graft were done in 28 cases each. RESULTS: Elimination of penile curvature, patient satisfaction and postoperative impotence rates were not statistically different for standard and modified Nesbit procedures. However, plaque excision and synthetic patch grafting resulted in less elimination of curvature (61%, p = 0.004), a lower rate of satisfaction (30%, p = 0.00002) and a higher incidence of impotence after surgery (18%, p = 0.04). The modified Nesbit procedure achieved an overall higher rate of correction of curvature than the standard approach (93 versus 79%). CONCLUSIONS: A modified Nesbit procedure achieves the greatest functional success for Peyronie's disease with an acceptably low complication rate. PMID- 9224324 TI - Comparison of puncture versus vasotomy techniques for vasography in an animal model. AB - PURPOSE: We determine the adverse effects of 2 different techniques of vasography in an animal model. MATERIALS AND METHODS: Unilateral vasography was performed by a direct puncture technique with a lymphangiogram needle or through a partial thickness vasostomy technique in 2 groups of 10 adults Lewis rats using nonionic contrast medium mixed with methylene blue. Each rat had a contralateral vasectomy. A complete vasogram was confirmed by visualization of colored dye in the bladder. An additional group of 5 animals with unilateral vasectomy alone served as controls. The adverse effects of these 2 techniques were assessed by performing mating studies at 2 and 4 months after vasography. In vitro flow through the vas deferens, sperm granuloma formation and histology of the vas deferens at the vasography site were evaluated at sacrifice 5 months after vasography. RESULTS: The fertility of the 3 groups, as measured by the mean number of uterine implantation sites, was not significantly different at the 2 and 4-month breeding periods. In addition, we observed no significant decrease in the fertility of the 3 groups with time. Complete vasal obstruction was noted at sacrifice in 2 rats (20%) in the vasostomy group and none in the puncture or control group (p = 0.476). The mean in vitro flow rates through the vasa of the puncture and vasostomy vasography groups were significantly lower than those in controls (p < 0.05) but not different from each other. The sperm granuloma formation rate was similar among the 3 groups. CONCLUSIONS: Our results demonstrate that both vasography techniques have a measurable adverse effect on vasal flow rates and a potential adverse effect on fertility. The direct puncture method had a slightly lower complication rate than the partial thickness vasostomy method and it may be the preferable technique for the inexperienced microsurgeon. PMID- 9224325 TI - Vasoepididymostomy for vasectomy reversal: a critical assessment in the era of intracytoplasmic sperm injection. AB - PURPOSE: We compared vasoepididymostomy to microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection for treatment of epididymal obstruction secondary to vasectomy. MATERIALS AND METHODS: Results in patients who underwent vasoepididymostomy for vasectomy reversal at our institution were compared to those reported previously for microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection performed for obstructive azoospermia. The pregnancy rates, delivery rates, complications, cost per procedure and cost per delivery were compared. A cost per newborn analysis was performed using pregnancy and delivery rates, and reported cost estimates for the complications of assisted reproductive techniques. RESULTS: A total of 55 men underwent 58 vasoepididymostomies in an attempt to restore fertility after vasectomy. Median followup was 19 months (range 0 to 115). Median obstructive interval was 12 years. There were no major complications. The patency rate after 6 months was 85%. Of the couples 20 achieved 24 pregnancies and 16 had 17 live births. The pregnancy rate at 1 year was 44%. There were 4 miscarriages and there are 3 ongoing pregnancies. The live delivery rate was 36%. Assuming a 29% delivery rate for microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection, the cost per newborn was $51,024, compared to $31,099 for vasoepididymostomy. CONCLUSIONS: Vasoepididymostomy is more successful and more cost-effective than microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection for vasectomy reversal. It does not expose the women to complications in the treatment of a male problem and it is indicated for treatment of epididymal obstruction secondary to vasectomy. Microsurgical epididymal sperm aspiration and intracytoplasmic sperm injection should be reserved for cases not amenable to surgical reconstruction. PMID- 9224326 TI - Effect of age at operation, location of testis and preoperative hormonal treatment on testicular growth after cryptorchidism. AB - PURPOSE: We evaluated the effect of patient age, primary location of the gonad and preoperative human chorionic gonadotropin administration on future testicular growth in patients treated for cryptorchidism. MATERIALS AND METHODS: Testicular volume was measured in 75 adults treated for cryptorchidism when they were 10 months to 13 years old. RESULTS: The mean volume of the cryptorchid testes plus or minus standard deviation, whether unilateral or bilateral, was 11 +/- 6 ml. compared to 20 +/- 7 ml. for the spontaneously descended testes in patients with unilateral cryptorchidism. The results showed no significant correlation between patient age at treatment or original testicular location and final testicular volume, although the 22 testes of 18 patients undergoing surgery after age 5 years were somewhat smaller (9 +/- 5 ml.) than the 66 testes of 55 younger patients (12 +/- 6 ml.). However, 26 patients who had received human chorionic gonadotropin treatment had a significantly smaller testis (9 +/- 5 ml.) than did 57 treated with surgery alone (12 +/- 6 ml., p < 0.05). CONCLUSIONS: Early orchiopexy at age younger than 2 years is not necessarily essential. Adult testicular volume is slightly greater in patients with cryptorchidism if treated at ages up to 5 years. Preoperative location of the testis in otherwise healthy boys exerts no definite effect on final testicular volume. Preoperative human chorionic gonadotropin administration may have an adverse effect on future testicular growth. PMID- 9224327 TI - Residual masses after chemotherapy for metastatic testicular cancer: the clinical implications of the association between retroperitoneal and pulmonary histology. Re-analysis of Histology in Testicular Cancer (ReHiT) Study Group. AB - PURPOSE: We determined the need and sequence of retroperitoneal lymph node dissection and thoracotomy in patients with nonseminomatous testicular cancer, and with residual retroperitoneal and pulmonary masses after chemotherapy. MATERIALS AND METHODS: We studied 159 patients undergoing retroperitoneal lymph node dissection and a thoracotomy following cisplatin based induction chemotherapy for metastatic testicular nonseminomatous germ cell tumor. Several well-known predictors for residual histology (necrosis, mature teratoma and cancer) were evaluated. RESULTS: As expected, necrosis was found more often at retroperitoneal lymph node dissection if the primary tumor was negative for teratoma, the residual mass was small or the decrease in size was great. Contrary, neither residual mass size nor the decrease in size was predictive of the histological status of the residual lung lesion. Histological findings in the retroperitoneum and lung were strongly correlated, such that necrosis at retroperitoneal lymph node dissection was associated with an 89% probability of necrosis in the lung. CONCLUSIONS: Retroperitoneal lymph node dissection should be performed before thoracotomy is considered, since the histological status at dissection is a strong predictor of that at thoracotomy. PMID- 9224328 TI - The impact of sonography on the diagnosis of scrotal disorders. AB - PURPOSE: We assessed the impact of ultrasound on the referring urologist diagnosis of scrotal disorders. MATERIALS AND METHODS: University urologists (6 attending and 3 resident physicians) in a clinic setting completed questionnaires before and after on 35 patients with scrotal symptoms. The physicians were requested to estimate the probability (0 to 100%) of their most likely diagnosis before and after receiving the sonographic information. We calculated the mean change in diagnostic percentage confidence and also the proportion of patients whose pre-sonographic diagnosis was changed. RESULTS: Scrotal ultrasound changed the initial diagnosis in 11 of 35 patients (32%, 95% confidence interval 19 to 49). The mean percentage gain in diagnostic certainty from ultrasound was 29% (95% confidence interval 20 to 39, p < 0.001). CONCLUSIONS: Scrotal ultrasound has considerable impact on referring urologist diagnosis of scrotal abnormalities. PMID- 9224329 TI - Natural history of prostatism: risk factors for acute urinary retention. AB - PURPOSE: We determined the occurrence of and risk factors for acute urinary retention in the community setting. MATERIALS AND METHODS: A cohort of 2,115 men 40 to 79 years old was randomly selected from an enumeration of the Olmsted County, Minnesota population (55% response rate). Participants completed a previously validated baseline questionnaire that assessed symptom severity, and voided into a portable urometer to measure peak urinary flow rates. A 25% random subsample underwent transrectal sonographic imaging of the prostate to determine prostate volume. Followup was performed through a retrospective review of community medical records to determine the occurrence of acute urinary retention in the subsequent 4 years. RESULTS: During the 8,344 person-years of followup 57 men had a first episode of acute urinary retention (incidence 6.8/1,000 person years, 95% confidence interval [CI] 5.2, 8.9). Among men with no to mild symptoms (American Urological Association symptom index score 7 or less) the incidence of acute urinary retention increased from 2.6/1,000 person-years among men 40 to 49 years old to 9.3/1,000 person-years among men 70 to 79 years old. By contrast, rates increased from 3.0/1,000 person-years for men 40 to 49 years old to 34.7/1,000 person-years among men 70 to 79 years old among men with moderate to severe symptoms (American Urological Association symptom index score greater than 7). Men with depressed peak urinary flow rate (less than 12 ml. per second) were at 4 times the risk of acute urinary retention compared with men with urinary flow rates greater than 12 ml. per second (95% CI 2.3, 6.6). Men with an enlarged prostate (greater than 30 ml.) experienced a 3-fold increase in risk (95% CI 1.0, 9.0, p = 0.04). CONCLUSIONS: Lower urinary tract symptoms, depressed peak urinary flow rates, enlarged prostates and older age are associated with an increased risk of acute urinary retention in community dwelling men. These findings may help to identify men at increased risk of acute urinary retention in whom closer evaluation may be warranted. PMID- 9224330 TI - A nationwide survey of practicing urologists: current management of benign prostatic hyperplasia and clinically localized prostate cancer. AB - PURPOSE: Our aim was to define the spectrum of urological care for benign prostatic hyperplasia (BPH) and clinically localized prostate cancer. MATERIALS AND METHODS: In 1995 a random sample of 394 American urologists was surveyed with a response rate of 67%. RESULTS: Respondents reported seeing a median of 240 BPH patients during the preceding 12 months, and they had prescribed alpha-blockers for 70 and finasteride for 15. They had performed a median of 25 transurethral prostatectomies but few other operations for BPH. Almost all urologists routinely used digital rectal examinations and prostate specific antigen tests for BPH diagnosis. The next most common studies were American Urological Association symptom scores and uroflowmetry. Pressure-flow studies were rarely done. Respondents reported seeing a median of 35 new patients with prostate cancer during the last year, and performing a median of 90 prostate biopsies and 13 radical prostatectomies. Respondents had referred a median of 10 patients for external beam radiotherapy but few patients received brachytherapy or cryotherapy. Urologist staging practices varied considerably. CONCLUSIONS: These data provide a picture of current practice regarding the management of BPH and prostate cancer. PMID- 9224331 TI - Reliability of Spanish translations of select urological quality of life instruments. AB - PURPOSE: Many patients with urological disease do not speak English. In medical studies restricting patients to those who speak only English undermines efforts to understand disease because restrictions decrease efficiency of patient recruitment, and because language and culture are associated with variable outcomes. In Spanish speaking locations, such as South Florida, studies would suffer severe selection bias if patients were required to speak English. To allow grouping in future studies of English and Spanish speaking patients we examined the English-Spanish reliability of select instruments that measure health related quality of life in patients with urological disease. MATERIALS AND METHODS: We assembled available Spanish versions and translated English versions of questions regarding satisfaction, the American Urological Association symptom index, the University of California, Los Angeles Prostate Cancer Index and a pain inventory. We then examined English-Spanish reliability by asking bilingual men 50 years old or older to complete English and Spanish versions at the same sitting. A convenience sample was recruited from outpatients and volunteers at the Miami Veterans Affairs Medical Center and population based subjects living in largely Hispanic Hialeah, Florida. Reliability estimates were calculated with kappa coefficients for categorical data and intraclass correlation coefficients for quantitative data. RESULTS: A total of 100 subjects a median of 59 years old completed the questionnaire, including 55 born in Puerto Rico or Cuba, while the remainder were born at various sites throughout the Americas and Spain. Reliability estimates showed that kappa = > 0.81 for almost all items. For 2 items relating to health and social interactions reliability was poor, and stratification showed that poor reliability was primarily a feature of subjects in good health who are theoretically socially active. CONCLUSIONS: Almost all items tested have excellent English-Spanish reliability in a mixed sample of bilingual men. Nonreliability of 2 items relating to health and social interactions probably originates from the effect of language on perception, and invalidates English and Spanish grouping of these items. Because the sample represents many dialects of Spanish, the translations tested may be transported to other cities. In studies that use these instruments investigators can reasonably group answers from English and Spanish speaking study subjects or study the effects of acculturation on quality of life. PMID- 9224332 TI - Transurethral resection of the prostate with microprocessor controlled electrosurgical unit. AB - PURPOSE: We analyzed the efficacy and side effects of a microprocessor controlled high frequency unit for transurethral resection of the prostate. MATERIALS AND METHODS: A high frequency device with microprocessor control was used in 934 consecutive patients undergoing transurethral resection of the prostate. Indications for transurethral resection, medical history, preoperative findings, operative parameters, operative and immediate postoperative complications, and postoperative peak flow rate and residual urine were evaluated. RESULTS: Postoperative peak flow rate and residual urine were comparable to those with standard transurethral resection of the prostate. One patient died on postoperative day 1 for a mortality rate of 0.1%. The immediate morbidity rate was 6.9%. CONCLUSIONS: Morbidity and mortality in this study were lower than those in previous series on transurethral resection of the prostate. PMID- 9224333 TI - Comparison of percent free prostate specific antigen and prostate specific antigen density as methods to enhance prostate specific antigen specificity in early prostate cancer detection in men with normal rectal examination and prostate specific antigen between 4.1 and 10 ng./ml. AB - PURPOSE: We analyzed the behavior of prostate specific antigen (PSA) density and percent free PSA to enhance the specificity of PSA in the early diagnosis of prostate cancer in men with normal digital rectal examination and PSA serum level between 4.1 and 10 ng./ml. MATERIALS AND METHODS: PSA serum level, PSA density and percent free PSA were analyzed in 74 men with normal digital rectal examination and PSA serum level between 4.1 and 10 ng./ml. All men underwent systematic prostate biopsy, and the diagnosis was benign prostate hyperplasia in 52 and prostate cancer in 22. Furthermore, we determined the decrease in unnecessary biopsies and the cancer detection rate using 0.10 versus 0.15 as cut points for PSA density, and 20 versus 25 as cut points for percent free PSA. RESULTS: In patients with benign prostatic hyperplasia and prostate cancer, respectively, the median PSA level was 6.7 and 7.0 ng./ml. (p > 0.05), median prostate volume was 50 and 37 cc (p < 0.04), median PSA density was 0.14 and 0.19 (p < 0.007) and median percent free PSA was 18.9 and 10.1 (p < 0.005). Using PSA density cut points of 0.15 and 0.10, the decrease in negative biopsies was 53.8 and 36.5% with a sensitivity of 86.4 and 90.9%, respectively. However, using percent free PSA cut points of 20 and 25, the decrease in negative biopsies was 36.5 and 26.9% with a sensitivity of 77.3 and 95.5%, respectively. CONCLUSIONS: Although both methods could minimize unnecessary biopsies in men with normal digital rectal examination and PSA serum level between 4.1 and 10 ng./ml., the percent free PSA was more cost-effective since transrectal ultrasound was not required. In this small series of symptomatic patients a percent free PSA cut point of 25 could detect at least 95% of prostate cancers and decrease 26.9% of negative biopsies. PMID- 9224334 TI - Prevalence and predictors of a positive repeat transrectal ultrasound guided needle biopsy of the prostate. AB - PURPOSE: We determined the prevalence of and risk factors for carcinoma in patients with 1 previously negative prostate biopsy. MATERIALS AND METHODS: Transrectal ultrasound guided prostate needle biopsies were repeated in 130 men. Risk factors analyzed included age, pathological result of initial biopsy, inter biopsy interval, prostate specific antigen (PSA), PSA density, PSA velocity, digital rectal examination, abnormal transrectal ultrasound and family history of prostate cancer. RESULTS: A total of 39 patients (30%) had positive biopsies for cancer. Univariate analysis revealed that PSA more than 20 ng./ml. and abnormal transrectal ultrasound were more frequent in men with positive second biopsies. Using multivariate logistic regression analysis only PSA more than 20 ng./ml. was a significant risk factor (adjusted odds ratio 4.48, 95% confidence interval 1.02 to 20.1). We determined the incidence of carcinoma in patients who represent the lowest risk group as defined by PSA less than 10 ng./ml., PSA density less than 0.15 mg./ml./cm.3, PSA velocity less than 0.75, ng./ml. per year, no prostatic intraepithelial neoplasia plus negative transrectal ultrasound, digital rectal examination and family history. Of 21 patients who fit this cohort 5 (23.8%) had carcinoma on repeat biopsy. CONCLUSIONS: A significant false-negative rate for initial transrectal ultrasound guided prostate biopsies exists. Baseline risk in lowest risk patients is sufficiently high such that one cannot define a subset of patients for whom repeat biopsy is unnecessary. We recommend repeat biopsy in all patients who meet the criteria for a transrectal ultrasound guided biopsy and in whom the initial biopsy is negative. PMID- 9224335 TI - p53 protein and gene alterations in pathological stage C prostate carcinoma. AB - PURPOSE: We determined the extent of p53 immunoreactivity in pathological stage C prostate cancer as well as its correlation to tumor grade, substage, recurrence and proliferation rate. To define better the temporal relationship of p53 nuclear reactivity in prostate cancer p53 immunoreactivity was evaluated in all associated prostatic intraepithelial neoplasia lesions. MATERIALS AND METHODS: Using immunohistochemistry p53 status and proliferation rate were determined in 96 tumors from patients with pathological stage C prostate cancer. Single strand conformational polymorphism in exons 5 to 8 was used in a subset of specimens to assess the association of p53 nuclear accumulation with mutations in the p53 gene. RESULTS: p53 Nuclear reactivity was demonstrated in 10 tumors (10.4%), including 6 with high and 4 with low level nuclear reactivity. Of the tumors 86 (89.6%) had no evidence of p53 immunoreactivity. Each of the 6 tumors with high level p53 reactivity had associated areas of prostatic intraepithelial neoplasia that also showed p53 nuclear reactivity. Furthermore, pathological stage C substage (C1, 2 or 3) was significantly associated with p53 nuclear reactivity (p = 0.04). Proliferation rates were correlated with p53 nuclear reactivity (p = 0.09), while there was no association with tumor grade or recurrence. p53 Gene alterations were noted in 2 of the 3 p53 positive tumors versus no alterations in the p53 gene of 3 p53 negative tumors. CONCLUSIONS: p53 Nuclear accumulation is uncommon in pathological stage C prostate cancer and its presence in premalignant prostatic intraepithelial neoplasia lesions suggests that it may be an early event in a subset of prostate cancers. PMID- 9224336 TI - Accelerated tumor proliferation rates in locally recurrent prostate cancer after radical prostatectomy. AB - PURPOSE: We compared the growth rates of primary cancer and prostatic fossa recurrence after radical prostatectomy. MATERIALS AND METHODS: Tumor proliferative rates were studied in 26 patients with biopsy proved prostatic fossa recurrences after radical prostatectomy. Proliferation was calculated in the prostatectomy specimens and in recurrent cancer using Ki-67 antibody to detect dividing cells. RESULTS: Mean and median labeling indexes for radical prostatectomy specimens were 2.96 and 2.51, respectively. Labeling indexes in locally recurrent tumors were significantly higher (mean 6.47, median 5.59, p < 0.001). The increase in labeling index between parent and recurrent tumors was unrelated to pathological staging at prostatectomy or interval from radical prostatectomy. CONCLUSIONS: Tumors that recur locally after radical prostatectomy appear to have a higher proliferative rate compared to parent tumors. PMID- 9224337 TI - The effect of urethral introducer tip catheters on the incidence of urinary tract infection outcomes in spinal cord injured patients. AB - PURPOSE: We attempted to determine whether an introducer tip catheter reduces urinary tract infection in spinal cord injured patients on intermittent catheterization. MATERIALS AND METHODS: The introducer tip catheter bypasses the colonized 1.5 cm. of the distal urethra. Enrolled patients were prospectively entered into the study in alternate groups depending on whether they reflex voided: group 1--on intermittent catheterization with the introducer tip catheter but not voiding spontaneously or wearing an external urinary catheter, group 2- same as group 1 but using a nonintroducer tip catheter; group 3--on intermittent catheterization with the introducer tip catheter, voiding by reflex and wearing an external urinary catheter, and group 4--same as group 3 but using a nonintroducer tip catheter. RESULTS: Statistical significance was shown when comparing patients using versus not using the introducer tip catheter regardless of whether an external urinary catheter was worn (p = 0.0121). A greater difference was noted between patients using and not using the introducer tip catheter in the intermittent catheterization only group (p = 0.0093). CONCLUSIONS: The introducer tip catheter decreased urinary tract infections in hospitalized men with spinal cord injury on intermittent catheterization. PMID- 9224338 TI - Extracorporeal shock wave lithotripsy for obstructing pancreatic duct calculi. AB - PURPOSE: A review was done to determine the effectiveness of extracorporeal shock wave lithotripsy (ESWL) in the treatment of impacted pancreatic duct calculi. MATERIALS AND METHODS: A total of 19 patients, who were potential candidates for radical pancreatic surgery after unsuccessful endoscopic retrograde cholangiopancreatography, sphincterotomy and attempted stone extraction from the pancreatic ducts, underwent ESWL of the calculi. Followup ranged from 6 months to 6 years. RESULTS: Of the 19 patients 14 avoided a major operation and 6 have remained pain-free for the long term. Two patients died of causes not related to ESWL or endoscopic retrograde cholangiopancreatography. Five patients eventually underwent a Whipple or Puestow procedure for relief of symptoms or persistent obstruction. Complications were minimal. CONCLUSIONS: ESWL is a valuable adjunct in patients with impacted pancreatic duct calculi unretrievable by primary endoscopic retrograde cholangiopancreatography. PMID- 9224339 TI - Use of polyglycolic acid mesh to support parenchymal closure following partial nephrectomy. AB - PURPOSE: We describe the use of absorbable mesh for closure of a large parenchymal defect created by partial nephrectomy. MATERIALS AND METHODS: A circular piece of polyglycolic acid mesh was measured to approximate the diameter of the parenchymal defect and the edges were sutured to the renal capsule in a running fashion. RESULTS: This technique has been used in 3 patients without complications. This approach has been particularly helpful for repairing large and irregular renal parenchymal defects. CONCLUSIONS: Use of polyglycolic acid mesh is effective for rapid, hemostatic closure of the kidney in association with partial nephrectomy. PMID- 9224340 TI - Endoscopic topical placement of formalin soaked pledgets to control localized hemorrhage due to radiation cystitis. AB - PURPOSE: A useful technique for treating localized bladder hemorrhage secondary to radiation cystitis is described. MATERIALS AND METHODS: Cotton pledgets soaked in 5% formalin are placed endoscopically onto bleeding foci of the bladder for 15 minutes and then removed. RESULTS: There was immediate cessation of prolonged bleeding refractory to intravesical saline, alum, prostaglandin E1 and estrogen. No subsequent bleeding was noted during 16 months of followup. CONCLUSIONS: Topical application of formalin soaked pledgets is an effective method of controlling localized bleeding secondary to radiation cystitis. PMID- 9224341 TI - Import catheter in erectile dysfunction. AB - PURPOSE: We propose an alternative technique for intracavernous self-injection of sodium nitroprusside for erectile dysfunction by inserting a Medtronic ImPort* catheter with a valved tip. MATERIALS AND METHODS: A silicone catheter was implanted in 3 patients with psychogenic impotence. The reservoir, which is used for vasoactive agent injection, was implanted laterally to the anterosuperior iliac spine and the distal tip of the catheter was inserted into the corpora cavernosa via a subcutaneous tunnel. The injection technique was taught to the patient and the initial injection was performed 1 week later. RESULTS: All patients and partners were satisfied with the technique and quality of erections at a mean followup of 14 months. There were no major local complications due to catheter implantation and no systemic complications due to sodium nitroprusside injection. CONCLUSIONS: An alternative technique for intracavernous pharmacotherapy of inserting an ImPort catheter prevented the complications of intracavernous injections in patients with erectile dysfunction. PMID- 9224342 TI - Retropubic cystourethropexy: is it an obstructive procedure? AB - PURPOSE: We sought to establish whether colpocystourethropexy creates bladder outlet obstruction, as evaluated by pressure-flow studies. MATERIALS AND METHODS: We retrospectively analyzed the records of 50 women. Preoperative evaluation included a detailed questionnaire, physical examination, urine culture, cystourethroscopy and multichannel urodynamic testing. Every patient underwent retropubic colpocystourethropexy according to the Tanagho modification of the original Burch technique. An average of 3 months after the operation clinical evaluation and identical multichannel urodynamic testing were repeated. Preoperative and postoperative urodynamic parameters were compared for each patient and statistical differences were established using Student's 2-tailed t test. RESULTS: No statistically significant difference was demonstrated in static urethral pressure profile parameters and in parameters during the filling phase of the cystometrogram except for cystometric capacity, which decreased after surgery (p = 0.02). In contrast, all 5 pressure-flow parameters analyzed (minimum urethral opening pressure, detrusor pressure at maximum flow, maximum flow, theoretical cross-sectional area and theoretical diameter of the flow rate controlling zone) showed statistically significant differences induced by surgery. Pressure-flow data reported on Schafer's diagram and on the Abrams Griffiths nomogram failed to demonstrate urodynamically significant obstruction created by surgery. CONCLUSIONS: Our data suggest that colpocystourethropexy does not create obstruction but, rather, restores pressure-flow conditions to a normal or nearly normal level. PMID- 9224343 TI - Urethral hemangioma: case report. PMID- 9224344 TI - Late glans hypervascularization subsequent to penile prosthesis implantation after revascularization operation. PMID- 9224345 TI - Treatment of impotence resulting from internal urethrotomy. PMID- 9224346 TI - Nonpalpable Leydig's cell tumors diagnosed by fine needle aspiration. PMID- 9224347 TI - Leech therapy for massive scrotal hematoma following percutaneous transluminal angioplasty. PMID- 9224348 TI - Papillary cystadenoma located in the spermatic cord. PMID- 9224349 TI - Finasteride induced gynecomastia. PMID- 9224350 TI - Re: Enucleative surgery for stage I nephroblastoma with a normal contralateral kidney. PMID- 9224351 TI - Re: Editorial comment: Outcome analysis of Mitrofanoff principle applications using appendix and ureter to umbilical and lower quadrant stomal sites. PMID- 9224352 TI - Re: Pelvic fracture urethral injuries: evaluation of various methods of management. PMID- 9224353 TI - Re: Circumcision: successful glanular reconstruction and survival following traumatic amputation. PMID- 9224354 TI - Re: The genetics of male infertility. PMID- 9224355 TI - Re: Japanese men have smaller prostate volumes but comparable urinary flow rates relative to American men: results of community based studies in 2 countries. PMID- 9224356 TI - Re: Characterization of patients with androgen independent prostatic carcinoma whose serum prostate specific antigen decreased following flutamide withdrawal. PMID- 9224357 TI - Marriage and mortality in prostate cancer. PMID- 9224358 TI - Re: Pathogenesis and prophylaxis of postoperative thromboembolic disease in urological pelvic surgery and Prospective randomized trial of warfarin and intermittent pneumatic leg compression as prophylaxis for postoperative deep venous thrombosis in major urological surgery. PMID- 9224359 TI - Urinary calcium oxalate saturation in healthy infants and children. AB - PURPOSE: A number of factors influence the development of renal calculi, the most essential of which is the supersaturation of urine with lithogenic substances. Calcium oxalate stones occur most frequently in adult and pediatric patients with urolithiasis. Therefore, we established normal age and sex related data for urinary calcium oxalate saturation in infancy and childhood to allow a more specific prediction of the risk of (recurrent) stone disease. MATERIALS AND METHODS: We collected 24-hour urine samples from 473 healthy infants and children without a history of renal stones. Urinary lithogenic and stone inhibitory substances were measured, and the urinary calcium oxalate saturation was calculated using a computer program. RESULTS: Mean urinary calcium oxalate saturation was always higher in boys than in girls, which was significant in infancy (5.22 versus 2.03, p < 0.05) and at ages 7 to 9 years (8.84 versus 5.47, p < 0.05). The saturation first increased (p < 0.05) until age 7 to 9 years in boys and girls, and remained at high levels at ages 10 to 12 years (7.03 versus 5.49, p < 0.05 compared to infancy). Calcium oxalate saturation then decreased until adolescence when values were comparable to those of infancy (5.29 versus 3.35). CONCLUSIONS: We recommend calculating urinary calcium oxalate saturation for diagnostic purposes as well as for therapy control. Normal age and sex related values must be considered. PMID- 9224360 TI - The ultrasonographic differentiation of obstructive versus nonobstructive hydronephrosis in children: a multivariate scoring system. AB - PURPOSE: We identified sonographic prognosticators to aid in distinguishing obstructive from nonobstructive hydronephrosis in children. MATERIALS AND METHODS: Twelve sonographic variables were initially analyzed to determine significant associations between the variables and the presence of urinary tract obstruction as defined by diuretic radionuclide renography. The significant findings were subsequently subjected to logistic regression models to identify potential predictors for obstructive hydronephrosis. RESULTS: The 7 variables associated with a significantly higher risk of urinary tract obstruction were increased echogenicity, parenchymal rims 5 mm. or less, contralateral hypertrophy, resistive index ratio 1.10 or greater, resistive index difference with diuresis of 70% or greater, ureter diameter 10 mm. or greater and aperistaltic ureter. These variables were used for the development of a multivariate scoring system. CONCLUSIONS: The obstructive scoring system shows promise as a screening method for the sonographic differentiation of obstructive from nonobstructive hydronephrosis in children. PMID- 9224361 TI - New renal scars in children with urinary tract infections, vesicoureteral reflux and voiding dysfunction: a prospective evaluation. AB - PURPOSE: Established renal scarring represents areas of the kidney that imaging reveals to be damaged at presentation for medical management of urinary tract infection. New renal scarring represents new renal damage in parts of the kidney that imaging reveals to be normal at presentation. We attempted to characterize patients in whom new renal scars developed while they were under our care. MATERIALS AND METHODS: In 1988 a data base was started to identify patients with new renal scarring. All patients presenting with urinary tract infections were enrolled. Our data base has 250 possible fields per event with multiple events per patient. More than 2,100 patients have been enrolled to date. All patients with pyelonephritis, defined as a febrile urinary tract infection with flank pain and tenderness, and all with reflux underwent dimercapto-succinic acid (DMSA) scan at least 4 months after presenting with infection to assess established renal scars. New renal scars were identified when new renal defects were demonstrated on a second DMSA scan. RESULTS: In our data base there are 1,426 patients with urinary tract infections, 685 (46%) with pyelonephritis and 1,062 (74.5%) with vesicoureteral reflux, including 558 found to have bilateral vesicoureteral reflux and 504 diagnosed with unilateral reflux. A history of daytime urinary incontinence was noted in 538 patients (37.7%), 192 (13.5%) had established scars at initial presentation and in 31 (2.1%) new renal scars developed while they were under our care, including 30 with established scars as well. Of the 25 patients in whom new renal scars developed while on medical therapy 11 underwent surgery. In 6 patients with dysfunctional voiding who were receiving medical treatment renal scars developed postoperatively. Surgery was performed in 17 of the 31 patients and 24 (77%) with new renal scars had a history of dysfunctional voiding. CONCLUSIONS: Previous characterizations of patients with new renal scars have relied on excretory urography for assessing renal architecture and ignored voiding patterns of the children affected. Using the DMSA scan we identified 31 children with reflux, urinary tract infection and dysfunctional voiding in whom new renal scars developed while they were under our care. PMID- 9224362 TI - Benign fibroepithelial polyps causing symptomatic bilateral intermittent hydroureteronephrosis. PMID- 9224363 TI - Bilateral congenital segmental megaureter. PMID- 9224364 TI - Left retrocaval ureter associated with the Goldenhar syndrome (branchial arch syndrome). PMID- 9224365 TI - Experience with vesicoureteral reflux in children: clinical characteristics. AB - PURPOSE: We reviewed our 9-year experience with a large population of children with vesicoureteral reflux who were evaluated and treated according to contemporary concepts. MATERIALS AND METHODS: From 1985 to 1993 we followed 288 boys and 752 girls with vesicoureteral reflux. If surgery was not performed, patients were on antibiotic prophylaxis and evaluation was done every 18 months with contrast voiding cystography and radionuclide renal imaging. Urine cultures were obtained every 4 months. Two negative voiding cystourethrograms 1 year apart were required to discontinue prophylaxis. RESULTS: The major reasons for initial evaluation were urinary tract infection in 560 children (54%), voiding dysfunction without urinary tract infection in 156 (15%), sibling surveys in 122 (12%) and prenatal hydronephrosis in 23 (2%). In 150 kidneys (10%) in 132 children scarring at presentation was grade 0 in 10 (7%), I in 18 (12%), II in 27 (18%), III in 30 (20%), IV in 48 (32%) and V in 17 (11%). Of these 132 patients 17 presented at ages less than 1 year (13%), 29 at ages 1 to 3 (22%), 50 at ages 4 to 6 (38%), 24 at ages 7 to 9 (18%) and 12 at ages greater than 10 (9%). No new scars were seen in children on prophylaxis without breakthrough infection. After 1 negative voiding cystourethrogram reflux was noted again in 27% of the cases. Breakthrough infections developed in 62 children of whom a third were older than 7 years. Reimplantation in 205 children (20%) was performed for grade IV to V reflux (101), breakthrough infection (62), advanced age (18), large periureteral diverticulum (12) and noncompliance (3). Five boys and 57 girls (30% of all children) had urinary tract infections after successful reimplantation. CONCLUSIONS: Almost half of the children with vesicoureteral reflux have no history of culture proved urinary tract infection. Scarring may be associated with any reflux grade and it may be initially diagnosed at any age. Only half of the scars are noted with higher grades of reflux (IV and V). Continuous prophylaxis prevents new scarring. Breakthrough infections are rare but they can occur at ages greater than 7 years. Two consecutive negative cystograms are necessary before discontinuing prophylaxis. Children should be monitored after reimplantation for recurrent urinary tract infection. PMID- 9224366 TI - Vesicoureteral reflux. PMID- 9224367 TI - Urodynamic correlates of resolution of reflux in meningomyelocele patients. AB - PURPOSE: Resolution of reflux in meningomyelocele patients is a reflection of improved bladder storage. We correlated resolution of reflux with changes observed in sequential urodynamic studies. MATERIALS AND METHODS: The study included 27 children with meningomyelocele born between 1975 and 1985 who presented with or developed vesicoureteral reflux. Resolution of reflux was observed during the 10-year followup period as they were treated with a regimen of clean intermittent catheterization and pharmaco-therapy. Urodynamic studies were performed when vesicoureteral reflux was present and subsequent to its resolution. The urodynamic parameters compared in the 2 studies included bladder capacity, pressure specific bladder volume, bladder compliance and leak point pressure. RESULTS: Significant increases in bladder capacity, pressure specific bladder volume and bladder compliance were noted. Leak point pressure appeared to be decreased subsequent to resolution of reflux. CONCLUSIONS: Resolution of reflux in meningomyelocele patients correlates with changes in parameters of bladder storage observed on sequential urodynamic studies. PMID- 9224368 TI - Bladder pheochromocytoma in a 10-year-old girl. PMID- 9224370 TI - Granulomatous lymphangitis of the penile skin as a cause of penile swelling in children. PMID- 9224369 TI - Urethral lengthening with anterior bladder wall flap (Pippi Salle procedure): modifications and extended indications of the technique. AB - PURPOSE: We report a clinical case series of an innovative method of urethral reconstruction for the treatment of urinary incontinence. Modifications of our original technique are presented. MATERIALS AND METHODS: Bladder neck repair was done in 17 patients, mean age 9.3, with neurogenic incontinence (13) or exstrophy (4). Average followup is 25.6 months. Of the patients 9 with neurogenic bladder underwent the original procedure using a midline anterior bladder wall flap. In the 4 patients with exstrophy a modified procedure was done using an anterolateral bladder wall flap. In another 4 patients an extended flap of distal mucosa was used to avoid ureteral reimplantation. Augmentation was performed in 13 of the 17 cases (10 detubularized ileum and 3 detubularized colon). RESULTS: Continence (greater than 4 hours) was obtained in 12 of the 17 patients (70%), 2 are dry for 1 to 2 hours and 3 are incontinent. A urethrovesical fistula developed in 2 patients (1 closed successfully), and 3 patients have problems with catheterization. CONCLUSIONS: Urethral lengthening with anterior bladder wall flap is a versatile alternative in the surgical treatment of urinary incontinence. Variations of the original technique resulted in an improved vascular supply and decreased the formation of fistula at the base of the flap. The modified technique was successful in patients with exstrophy, including those who had failed bladder neck surgery. PMID- 9224371 TI - Epidermoid cyst of the penis with extension into the pelvis. PMID- 9224372 TI - Endocrine analysis of childhood monorchism. AB - PURPOSE: We characterized follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels in boys with surgically documented unilateral absent testes (monorchism) to determine whether measurement of gonadotropin levels could distinguish them from boys with unilateral impalpable cryptorchidism. MATERIALS AND METHODS: Baseline serum gonadotropin levels were prospectively measured in 43 boys 2 months to 14 years old who presented with a unilateral impalpable testis that was confirmed to be absent at surgery. Control serum specimens were obtained from 63 age matched boys undergoing minor surgery with no evidence of hypospadias, or testicular, hormonal or renal diseases. Serum FSH and LH levels were drawn preoperatively and assayed by double antibody radioimmunoassay. A subgroup of 7 boys with monorchism was also evaluated following gonadotropin releasing hormone (GnRH) stimulation and compared to age matched boys with a unilaterally impalpable testis discovered surgically. RESULTS: In the monorchism group mean plus or minus standard deviation basal FSH was 4.08 +/- 0.28 mIU/ml. and LH was 4.13 +/- 0.33 mIU/ml. In the control group mean basal FSH was 4.36 +/- 1.52 mIU/ml. and LH was 4.66 +/- 0.75 mIU/ml. No statistical difference existed between the 2 groups for mean basal gonadotropin level. While monorchid boys were more likely to have elevated FSH levels (p = 0.016), this was not true for LH (p = 0.21). Since gonadotropin levels less than 5 mIU/ml. are accepted normal values, this threshold was applied to FSH and carried a sensitivity of 23.8%, specificity 93.8%, positive predictive value 71.4% and negative predictive value 65.6%. Lower cutoff values marginally improved sensitivity but reduced specificity. Peak stimulated levels of FSH and LH following GnRH stimulation failed to distinguish between boys with 1 or 2 testes. CONCLUSIONS: Baseline FSH is more likely to be elevated in prepubertal boys with monorchism but it does not appear to be clinically useful when sensitivity and predictive value are poor. Similarly, gonadotropin level following GnRH stimulation is not sufficiently sensitive to advocate the use of hormonal measurements to diagnose prepubertal monorchism. PMID- 9224373 TI - Bilateral suprarenal cryptorchidism in a patient with the Pfeiffer syndrome. PMID- 9224374 TI - Testicular exstrophy: bilateral presentation in a newborn. PMID- 9224375 TI - Cystic dysplasia of the rete testis: a benign congenital lesion associated with ipsilateral urological anomalies. AB - PURPOSE: Cystic dysplasia of the rete testis is a benign congenital lesion that can mimic testicular cancer. We report 6 cases, review the literature, discuss the embryological etiology and make management recommendations. MATERIALS AND METHODS: The records and pathology reports of 6 boys presenting with cystic dysplasia of the rete testis at 5 institutions were reviewed, as was the relevant literature. RESULTS: Of the 6 cases 5 presented as scrotal masses in previously healthy boys and 1 as an abdominal mass in a newborn with multiple congenital anomalies. One patient had been followed from birth for a multicystic dysplastic kidney and 4 were found to have an ipsilateral absent kidney during evaluation. Development of the contralateral side was normal in most cases. CONCLUSIONS: Cystic dysplasia of the rete testis is an unusual, benign congenital lesion that can mimic testicular cancer in presentation. The presence of ipsilateral renal anomalies, particularly renal agenesis, can suggest cystic dysplasia of the rete testis in the differential diagnosis preoperatively. Even if cystic dysplasia of the rete testis is suspected, we recommend inguinal exploration and early control of the spermatic cord in the event that neoplasia is identified. If possible, the goal of preserving as much normal testicular parenchyma as possible is desirable. Long-term followup for possible recurrence is recommended, particularly after local excision. PMID- 9224376 TI - Ipsilateral testicular hypotrophy is associated with decreased sperm counts in infertile men with varicoceles. AB - PURPOSE: The presence of ipsilateral testicular growth retardation (hypotrophy) is the most common indication for prophylactic varicocele repair in adolescents in an effort to prevent future infertility. We examined the relationship between semen parameters and ipsilateral versus contralateral testicular size in men with unilateral varicoceles to determine whether testicular size is an appropriate parameter for predicting future fertility. MATERIALS AND METHODS: We studied the records of consecutive patients with palpable unilateral left varicoceles for whom a history, physical examination and semen analysis were available. Total motile sperm counts of men with and without ipsilateral testicular hypotrophy were compared. RESULTS: We identified 611 patients with unilateral clinical left varicoceles, including 305 (50%) with ipsilateral testicular hypotrophy. Mean total motile sperm counts plus or minus standard error of mean were significantly less in the patients with than without testicular hypotrophy (80 +/- 5.2 versus 126 +/- 7.8 x 10(6) sperm, p = 0.0018). Hypotrophy was more common in patients with large varicoceles (73%) than in those with medium (53%) or small (43%) varicoceles. CONCLUSIONS: Infertile patients with testicular hypotrophy associated with unilateral varicoceles have worse semen parameters than those without hypotrophy. These data support the practice of varicocele repair in adolescents with varicocele associated testicular growth retardation. PMID- 9224377 TI - Malignant Sertoli cell tumor in a prepubescent boy. PMID- 9224378 TI - Serum levels of mullerian inhibiting substance in preterm and term male neonates. AB - PURPOSE: Mullerian inhibiting substance, also called anti-mullerian hormone, is responsible in the embryo for the regression of the mullerian structures. During the second and third trimesters the physiological functions that mullerian inhibiting substance may have after the period of mullerian duct regression are poorly understood. We obtained information on mullerian inhibiting substance levels in the male newborn during this period of gestation. MATERIALS AND METHODS: Mullerian inhibiting substance was measured by an enzyme immunoassay in cord blood obtained at birth in 27 preterm (25 to 36 weeks of gestation) and 92 term (37 to 42 weeks) male neonates. RESULTS: Cord serum mullerian inhibiting substance concentrations were relatively high from 25 to 31 weeks (mean plus or minus standard deviation 86.4 +/- 36.1 ng./ml.) and then they decreased from 32 weeks to term (mean 24.2 +/- 14.0 ng./ml.). CONCLUSIONS: The decline early in the third trimester may be consistent with mullerian inhibiting substance having a function during the second but a diminished role in the third trimester. PMID- 9224379 TI - Testicular descent--a proposed interaction between mullerian inhibiting substance and epidermal growth factor. PMID- 9224380 TI - Immunohistochemical expression of Ki-67 to predict lymph node involvement in clinical stage I nonseminomatous germ cell tumors. AB - PURPOSE: Primary archival tumor tissues of 89 patients with clinical stage I nonseminomatous germ cell tumors were analyzed for MIB-1 expression and histological parameters such as percentage embryonal carcinoma and presence of vascular invasion to determine the value of these parameters to predict absence or presence of occult lymph node disease. MATERIALS AND METHODS: A monoclonal antibody (MIB-1) developed for application in paraffin-embedded tissues was used to measure immunohistochemical expression of Ki-67 for the overall tumor (total MIB-1) and for each malignant cell type present. In addition, the primary tumors were examined for the presence of vascular invasion and determination of quantitative histology. Univariate and multivariate logistic regression models were used for statistical analysis. RESULTS: Univariate analysis neither revealed total MIB-1 score nor MIB-1 score in the highest area of staining of the different cell types to significantly predict pathological stage I or stage II disease. However, the presence of vascular invasion (p < 0.0001) and the percentage of embryonal carcinoma (p < 0.0001) were significant risk factors for occult nodal disease. Multivariate logistic regression analysis revealed the combination of vascular invasion and the percentage of embryonal carcinoma to be the best model to predict pathological stage II correctly (86.5%). DISCUSSION: The determination of immunohistochemical MIB-1 expression did not correlate with pathological stage in clinical stage I nonseminomatous germ cell tumors (NSGCT). We were not able to define high risk or low risk groups for occult nodal disease based on MIB-1 staining results. However, percentage of embryonal carcinoma and presence of vascular invasion accurately predicted absence or presence of lymph node metastasis in clinical stage I NSGCT. Our study underlines that a prospective multicenter trial is urgently needed to accurately assess the role of MIB-1 staining in management of clinical stage I NSGCT. PMID- 9224381 TI - Apoptosis in the rat penis after penile denervation. AB - PURPOSE: Despite the advances in nerve sparing prostatectomy for prostate cancer, some patients develop impotence or subjectively complain of a decrease in penile size. We hypothesized that these clinical observations may be explained by injury to the cavernous nerves resulting in programmed cell death (apoptosis) within the penis. We utilized a rat model of penile denervation in order to demonstrate apoptosis after denervation. METHODS AND MATERIALS: Fifteen male Sprague Dawley rats underwent abdominal exploration and bilateral cavernous neurotomy. Fifteen sham operations were performed as normal controls. The rats were sacrificed on postoperative day 1,2,3,6, and 10 and their penises were harvested. Messenger RNA was extracted and probed on a northern blot for sulfated glycoprotein-2 (SGP-2). SGP-2 is a gene product reported to be elevated in apoptotic tissues. Separate denervated and sham rats were used for DNA extraction (sacrificed postoperative day #2) in order to demonstrate the internucleosomal DNA fragmentation (laddering) found in apoptotic tissues. In addition, in situ histology was performed with ISEL techniques (in situ end labeling) to stain for apoptotic nuclei in denervated rats. RESULTS: Northern blot analysis showed a large increase in SGP-2 mRNA expression in the denervated rats with little detected in the sham operated group. DNA extraction studies revealed the presence of internucleosomal DNA fragmentation on agarose gel (a marker for apoptosis) in the denervated group versus intact high molecular weight DNA in the sham rats. In addition, in situ staining of denervated penile erectile tissue demonstrated apoptotic nuclei in the cavernous tissue. CONCLUSION: Apoptosis of penile erectile tissue occurs after denervation of the rat penis. This has not been previously described in the literature and may offer some explanation at the molecular level concerning the mechanism of impotence and/or decrease in penile size after radical prostatectomy. PMID- 9224382 TI - In vivo cystometric evaluation of progressive bladder outlet obstruction in rats. AB - PURPOSE: Bladder outlet obstruction in man is a common medical disorder that may result from benign prostatic hyperplasia, urethral stricture disease, or congenital anomaly. The functional changes that develop in response to obstruction include detrusor instability, elevated voiding pressures, and the presence of a residual urine. The aim of this study was to document the development of progressive bladder outlet obstruction over time in a rat model using conscious, in vivo urodynamics. MATERIALS AND METHODS: Infravesical bladder outlet obstruction was created in female rats by placing a jeweler's jump ring loosely around the proximal urethra. Gradual development of outlet obstruction was followed urodynamically in awake animals at 3, 7, 14, 21, and 28 days post obstruction using a subcutaneously implanted mediport. For each group n = 5-8 animals. RESULTS: Animals developed large capacity bladders with increased compliance, a high residual urine volume, and spontaneous activity. Bladder capacity increased from 0.20 + 0.02 ml. to 6.30 + 1.59 ml. at 28 days post obstruction (p < 0.05). Residual volume increased from 0.06 + 0.01 ml. to 5.95 + 1.54 ml. (p < 0.05). Percent void decreases from 72 + 3.7% in sham controls to 6.7 + 2.5% at 28 d (p < 0.05). Voiding pressure increased from 12 + 1.6 mm. Hg in sham animals to a maximum of 42 + 6.1 mm. Hg at 21 d (p < 0.05). Compliance was significantly higher at 28 d when compared to all other time points. 89% of obstructed animals developed bladder instability. CONCLUSIONS: This study provides clear evidence of the progressive change in bladder function which occurs following outlet obstruction. Implantation of a subcutaneous mediport allows in vivo recording of both the filling and voiding phases of micturition in awake animals that have intact neural responses. This is a precise and easily reproducible method for producing obstruction in a small animal which can provide a continuum of tissue and urodynamic data that may be used to further study the pathophysiologic changes underlying bladder outlet obstruction or other models of bladder dysfunction. PMID- 9224383 TI - Cytotoxic effects of recombinant adenovirus p53 and cell cycle regulator genes (p21 WAF1/CIP1 and p16CDKN4) in human prostate cancers. AB - PURPOSE: Overexpressing or restoring the basal levels of tumor suppressor genes in cancer cells can suppress tumorigenicity of cancer cells. In this communication, we compared tumor suppressive activities of three well-defined tumor suppressive genes (p53, p21WAF1/CIP1, and p16CDKN2) delivered individually to prostate cancer cells with adenoviral vector (Ad). MATERIALS AND METHODS: Efficacy of growth inhibition by recombinant adenoviruses bearing p53, p21WAF1/CIP1, or p16CDKN2 (Ad5CMV-p53, Ad5CMV-p21, Ad5CMV-p16) genes were tested in vitro on androgen-dependent (LNCaP) and androgen-independent (C4-2, DU-145, and PC-3) human prostate cancer cells, ex vivo and in vivo on PC-3 tumor. RESULTS: Ad5CMV-p53 was observed to exert the greatest growth inhibitory action on all of the cell lines tested; inhibition appeared to be cytolytic. In comparison to control Ad5CMV-PA added samples, the growth inhibitory action of Ad5CMV-p21 and Ad5CMV-p16 appeared to be cytostatic. Ad5CMV-p53 is more effective than Ad5CMV-p16 and Ad5CMV-p21 in inhibiting the tumor "take" rate. A similar order of antitumor activity was observed when recombinant adenoviruses were injected intratumorily to previously established PC-3 tumors in vivo. CONCLUSION: p53 is the most effective tumor suppressor gene to target human prostate cancer. PMID- 9224384 TI - Results of laser tissue soldering in vasovasostomy and epididymovasostomy: experience in the rat animal model. AB - The use of microsurgical techniques (vasovasostomy and epididymovasostomy) for vasectomy reversal has now enabled surgeons to perform both procedures with certainly acceptable success rates. However, these operations are technically demanding and require special training in microsurgery. PURPOSE: A new method of performing these procedures using laser tissue soldering is described and results are evaluated. Laser tissue soldering is different from laser welding in that it involves the laser activation of a protein solder with a dye specific for the specific wavelength of laser light; therefore, surrounding tissue is not affected by the laser. MATERIALS AND METHODS: Ten rats underwent bilateral vasovasostomy and eleven underwent bilateral epididymovasostomy. In each rat, a sutured anastomosis was performed on one side while laser tissue soldering was performed on the other. Animals were sacrificed after one month and anastomoses were evaluated for patency and presence of sperm granulomas. Histologic analysis was also performed. RESULTS: Patency rates were 8/10 (80%) for sutured vasovasostomy versus 9/10 (90%) for the laser technique. Epididymovasostomy patency rates were 8/11 (73%) for sutured versus 9/11 (82%) for the laser technique. Mean operative times were significantly shorter for lasered anastomoses when compared to controls. The frequency of granuloma formation did not significantly differ between laser and control groups. CONCLUSIONS: Laser tissue soldering resulted in similar patency when compared to a conventional 2 layered sutured anastomosis while decreasing operative time. In addition, since fewer sutures are placed, the laser method is less technically demanding. PMID- 9224385 TI - Genetically regulated response to intravesical bacillus Calmette Guerin immunotherapy of orthotopic murine bladder tumor. AB - PURPOSE: Genetically regulated host response to intravesical Bacillus Calmette Guerin (BCG) immunotherapy was assessed using the murine bladder tumor MM45T in Bcgr and Bcgs inbred congenic strains of mice. MATERIALS AND METHODS: Tumor detection and monitoring of treatment response to BCG was carried out using magnetic resonance imaging (MRI) of BALB/c (Bcgs allele) and BALB/c. CD2 (CD2) (Bcgr allele) mice implanted orthotopically with MM45T tumor cells. Intravesical BCG instillation (3 doses per week for 3 weeks) was used as prophylaxis against tumor implantation in both Bcgr and Bcgs strains and as definitive treatment against MRI-confirmed established tumors. Tumors implanted in both strains of untreated mice served as controls. Intravesical injection of BCG was also performed in established heterotopic subcutaneous tumors in both strains. Immunologic response in all groups was assessed by flow cytometric analysis of the bladder irrigation fluid cell composition, measuring CD4+ (helper/inducer) and CD8+ (cytotoxic/ suppressor) cell subsets. RESULTS: Intralesional injection of BCG into established heterotopic tumors showed growth inhibition in the Bcgs strain but not in the Bcgr strain. Intravesical BCG treatment against established orthotopic tumors showed significant tumor regression in the Bcgs strain compared to control but there was no effect in the Bcgr strain. CONCLUSION: The differential anti-tumor activity of BCG in the Bcgs and Bcgr congenic murine strains supports the notion that Bcg gene-controlled responsiveness to BCG innoculation determines, at least partially, the host response to immunotherapy. These results have potential clinical significance in patient selection for intravesical therapy for bladder cancer. PMID- 9224386 TI - Renin angiotensin system in rabbit corpus cavernosum: functional characterization of angiotensin II receptors. AB - PURPOSE: The regulation of the corporal smooth muscle tone is important in the process of penile erection. Although specific angiotensin (ANG) II binding to and effects of ANG II on some reproductive structures have been studied, the presence of the renin-angiotensin system has not yet been defined in the corpus cavernosum. ANG II is formed from ANG I by angiotensin I-converting enzyme (ACE). ANG II and ANG I produce contractions in vascular smooth muscles. Two subtypes of ANG II receptors (AT1 and AT2) have been characterized. The purpose of the present experiments was to determine whether the renin-angiotensin system regulates rabbit corpus cavernosum smooth muscle tone. MATERIALS AND METHODS: A strip of rabbit corpus cavernosum was mounted in an organ chamber to measure the isometric tension. The specific binding for 125I-ANG II was characterized by in vitro autoradiography. RESULTS: ANG II and ANG I, precursor of ANG II, contracted corpus cavernosum smooth muscle dose-dependently, but the response of smooth muscle to ANG I was 10-fold less than that to ANG II. Contractile responses of smooth muscle to ANG II and ANG I were blocked by Dup 753, a specific inhibitor of ANG II type 1 receptor, but not by PD 123,319, a specific inhibitor of ANG II type 2 receptor. The effect of ANG I was attenuated by captopril, an inhibitor of ACE. Specific binding sites for 125I-ANG II were found in the corpus cavernosum. The dissociation constant (Kd) was 5.32 +/- 1.65 nM. and maximum binding capacity (Bmax) was 305.72 +/- 85.24 amol/mm. Specific binding of 125I-ANG II was displaced by Dup 753 (10(-6) M) but not by PD 123,319 (10(-5) M). The inhibitory constant (Ki) for Dup 753 was 8.09 +/- 2.51 nM. CONCLUSION: The present results suggest that the renin-angiotensin system is involved in the regulation of corpus cavernosum smooth muscle tone of rabbit and the ANG II receptor subtype AT1 is important in the regulation of penile erection. PMID- 9224387 TI - Cu/Zn superoxide dismutase, catalase and glutathione peroxidase mRNA expression in the rat testis after surgical cryptorchidism and efferent duct ligation. AB - The testis is known to be highly sensitive to a number of physical stresses. Previous investigations suggest that oxidative stress may be an important mediator of testicular injury. The ability of the testis to manage oxidative stress may be limited by enzymatic clearance of reactive oxygen species (ROS). To evaluate the ability of the testis to withstand the common pathologic conditions of cryptorchidism and obstruction, we measured mRNA levels of testicular antioxidant enzymes. Prepubertal rats were rendered unilaterally cryptorchid and 40 days after the procedure, cryptorchid, contralateral and control (sham) testes were harvested for RNA extraction. Adult rats were subjected to unilateral efferent duct ligation and the obstructed testes harvested 1 to 28 days after the procedure. Antioxidant enzyme mRNA expression was assessed by Northern blot analysis using 32P-labeled DNA probes for classical cellular glutathione peroxidase (GSHPx), phospholipid hydroperoxide glutathione peroxidase (PHGPX), Cu/Zn superoxide dismutase (SOD) and catalase. In both cryptorchid and contralateral testes, the germ cell-specific 0.9 kb SOD and PHGPX mRNA transcript levels were significantly decreased compared to control testes (p < 0.05). Similarly, after efferent duct ligation, the 0.9 kb SOD and PHGPX mRNA transcript levels also decreased compared to control testes (p < 0.05). These findings suggest that the overall decline in testicular mRNA transcript levels after efferent duct ligation and cryptorchidism is primarily due to germ cell depletion. Reduced levels of antioxidant enzyme mRNAs in cryptorchid testes have been documented. Further experiments may elucidate the role of increased oxidative stress associated with decreased antioxidants in cryptorchidism. It remains to be determined whether oxidative stress has a causative role in the abnormal spermatogenesis and tumorigenesis associated with cryptorchidism. PMID- 9224388 TI - Effects of magnesium cardioplegia on regulation of the porcine coronary circulation. AB - We compared the effect of hypermagnesium and hyperkalemic crystalloid cardioplegia on beta-adrenoceptor-mediated and endothelium-dependent relaxation and myogenic responses of coronary arterioles. Pigs were placed on cardiopulmonary bypass. Hearts were arrested with cold hypermagnesium (25 mM Mg2+, hyper-Mg, n = 12) or hyperkalemic (25 mM K+, hyper-K, n = 12) crystalloid cardioplegia for 1 hr. Hearts of selected pigs (n = 6 in each group) were then reperfused for 1 hr. In vitro relaxation responses of acetylcholine-pre contracted arterioles were studied in a pressurized no-flow state with video microscopy. Relaxation of pre-contracted coronary microvessels (70-150 microns) to isoproterenol (beta-adrenergic agonist) and forskolin (adenylate cyclase activator) was preserved after cardioplegia using a hyper-Mg solution. In contrast, responses were impaired to isoproterenol [P < 0.01 (two-factor ANOVA) vs. controls, n = 6] and forskolin (P < 0.01) after hyper-K cardioplegia. After 1 hr of reperfusion, relaxation responses to isoproterenol and forskolin were partially recovered. Hyper-Mg cardioplegia and post-cardioplegic reperfusion did not affect receptor-mediated endothelium-dependent relaxation to ADP, non receptor-mediated endothelium-dependent relaxation to A23187, and endothelium independent relaxation to nitroprusside. However, responses to ADP (P < 0.01) and A23187 (P < 0.05) were selectively impaired after hyper-K cardioplegia. Myogenic contraction was impaired after either hyper-Mg or hyper-K cardioplegia. Left ventricular systolic pressure, coronary blood flow, and +dP/dt were similar after hyper-Mg or hyper-K cardioplegia. These results suggest that hyper-Mg cardioplegia is superior to hyper-K cardioplegia in terms of preserving beta adrenoceptor-mediated and endothelium-dependent regulation of the coronary microcirculation, yet it has minimal if any additional beneficial effect on preserving myogenic responses or myocardial contractile function. PMID- 9224389 TI - Lexipafant inhibits platelet activating factor enhanced neutrophil functions. AB - Platelet activating factor (PAF) enhances polymorphonuclear leukocyte (PMN) superoxide (.O2-) production, CD11b expression, and elastase release, all essential components in the pathophysiology of multiple-organ failure. This study was designed to determine the effects of Lexipafant, a PAF receptor antagonist, on PAF-mediated PMN functions. PMNs from 10 healthy volunteers were isolated and pretreated with various concentrations of Lexipafant (0-100 microM). PMNs were then incubated for 5 min with 200 nM PAF for .O2- detection or 2000 nM PAF for elastase measurement and activated with 1 microM N formylmethionylleucylphenylalanine. The mean rate of .O2- production was determined by a cytochrome c reduction assay (nmole .O2-/min/1.33 x 10(5) PMN +/- SEM). Elastase release was measured by the cleavage of the synthetic elastase substrate Meo-Suc-Ala-Ala-Pro-Val-pNA (mean elastolytic activity +/- SEM). In parallel experiments, PMNs were incubated with 200 nM PAF for 30 min following pre-treatment with Lexipafant and CD11b expression was determined by flow cytometry (mean fluorescence intensity +/- SEM). Statistical analysis was performed using repeated-measures ANOVA (P < 0.05). Lexipafant inhibited PAF enhanced PMN .O2- generation, CD11b expression and elastase release in a dose dependent fashion. The IC50 of Lexipafant for .O2- production, CD11b expression, and elastase release was 0.046, 0.285, and 0.05 microM, respectively. Lexipafant attenuated the PAF-mediated upregulation of PMN .O2- production, CD11b expression, and elastase release in a dose dependent fashion. These data support the hypothesis that Lexipafant may reduce the severity of the inflammatory response to injury produced by PAF-enhanced activation of PMNs. PMID- 9224390 TI - Immunization with antibodies that mimic LPS protects against gram negative bacterial sepsis. AB - We developed 9H1.B11, an anti-idiotypic anti-deep core/lipid A (DCLA), murine monoclonal antibody (mAb) that mimics the conserved DCLA region of lipopolysaccharide (LPS). It recognizes an epitope in the variable region of an DCLA mAb, binds to the murine macrophage cell surface, and inhibits LPS-induced macrophage cytokine secretion. We hypothesized that (1) active immunization with mAb 9H1.B11 would be associated with the development of anti-DCLA antibodies and (2) immunization would protect against subsequent gram negative bacterial challenge. Mice were immunized for 8 weeks before intraperitoneal (ip) challenge with Escherichia coli O111:B4 bacteria. Control animals were immunized with an irrelevant IgM antibody 8133 (negative control) or with LPS derived from Salmonella minnesota Re bacteria (positive control). Sera from immunized mice were collected, and titers against the core region of LPS (Re) and against LPS derived from the infecting E. coli strain were determined. Mice immunized with mAb 9H1.B11 developed measurable titers against S. minnesota Re LPS but not against the challenge strain of E. coli. However, immunization with 9H1.B11 on S. minnesota Re LPS protected against subsequent infection due to E. coli O111:B4 (100% survival). The group of mice immunized with IgM 8133 exhibited only 25% survival. The development of an anti-S. minnesota Re LPS titer after immunization with 9H1.B11 provides further evidence that a portion of 9H1.B11 mimics the conserved DCLA region of LPS. We believe that this approach holds considerable promise and plan further studies to define the mechanism by which protective capacity occurs. PMID- 9224391 TI - Anastomotic intimal hyperplasia: a comparison between conventional and endovascular stent graft techniques. AB - Endovascular grafts (EVGs) have been proposed as a treatment for a variety of vascular diseases; however, the impact of EVGs on graft healing has not been fully evaluated. The aim of this study is to compare anastomotic intimal hyperplasia (AIH) and endothelialization in EVGs and conventional bypass grafts (CGs). Seven mongrel dogs received an EVG in one iliac artery and a CG in the other iliac artery using a 5 mm x 4 cm polytetrafluoroethylene graft. The EVG was secured to the native vessel wall, with balloon expandable stents at either ends of the graft. CGs were anastomosed using running sutures. Intravascular ultrasound was performed at the time of sacrifice (8 weeks) to determine percentage of stenosis at the distal anastomosis. Specimens were divided longitudinally for light microscopic analysis (thickness of distal AIH) and scanning electron microscopic studies (percentage of endothelial coverage of the graft). Percentage of stenosis at the distal anastomosis was significantly higher in EVGs compared with CGs (28.2 +/- 18.2% versus 1.8 +/- 2.8%; P < 0.01) due to significantly greater mean intimal thickness in the EVGs (441.1 +/- 101.1 microns versus 82.4 +/- 41.9 microns; P < 0.01). The total percentage of area covered by endothelial cells was also significantly greater in EVGs compared with CGs (80.5 +/- 37.5% versus 30.3 +/- 37.1%; P < 0.05). Intraluminal location enhanced endothelialization of the polytetrafluoroethylene graft; however, it also resulted in greater AIH. Further device refinements including stent design may be required to maximize the potential of these endovascular procedures. PMID- 9224392 TI - Alpha-adrenergic activation of myocardial NF kappa B during hemorrhage. AB - Hemorrhage and resuscitation has been recognized as an exclusively destructive process which results in multiple organ dysfunction. Although it is well established that endogenous adaptation (preconditioning) mechanisms exist, it is unknown whether hemorrhage and resuscitation induces endogenous adaptive/protective mechanisms in the heart. Furthermore, alpha 1-adrenoceptors and nuclear factor kappa B (NF kappa B) have each been implicated in stress induced signal transduction; however, whether they might be involved in hemorrhage-induced adaptive signal transduction remains unknown. This study tests the hypothesis that H/R activates myocardial NF kappa B and results in myocardial adaptation via alpha 1-adrenoceptors. Rats were briefly (10 min) hemorrhaged to 35 mmHg and resuscitated, sham operated, or neither, with and without prior alpha 1-adrenoceptor inhibition (prazosin). Hearts were then isolated and either probed for NF kappa B activation or subjected to a second insult consisting of global normothermic I/R (20 min/40 min). Antecedent hemorrhage and resuscitation activated myocardial NF kappa B and improved left ventricular developed pressure, coronary flow, and end diastolic pressure following ischemia-reperfusion (P < 0.05, ANOVA with Bonferroni-Dunn). Hemorrhage-induced adaptation was abolished by prior alpha 1-adrenoceptor blockade. This study constitutes the initial demonstration that H/R activates myocardial NF kappa B and induces adaptive signal transduction against ischemia-reperfusion injury. PMID- 9224393 TI - Expression of two novel recombinant proteins from aortic adventitia (kappafibs) sharing amino acid sequences with cytomegalovirus. AB - We have recently purified and partially sequenced a microfibrillar protein from human aortic adventitia (aneurysm-associated antigenic protein, 40 kDa [AAAP-40]) that is immunoreactive with immunoglobulin (IgG) from the wall of abdominal aortic aneurysms (AAAs). It shares motifs with Ig kappa (which may act as a binding site for interaction with integrins), cytomegalovirus (which may be a molecular mimic in the pathogenesis of AAA), and vitronectin and the fibrinogens. A cDNA library was constructed from the aortic adventitia of a AAA. The library was screened with either rabbit anti-vitronectin antibody or rabbit anti fibrinogen antibody. Positive plaques were purified and expressed in a strain of Escherichia coli. The clone sequences were analyzed. The expressed proteins were separated by SDS/PAGE and the immunoblots were probed with either AAA IgG or anti human Ig kappa antibody. Experimental cell lines, transfected with the clones (clones 1 and 5), synthesized recombinant proteins (rAAAP-CL1 and rAAAP-CL5), detectable in Western immunoblots with AAA IgG. A prediction of the tertiary structure resembles well-characterized cell adhesion molecules. These findings suggest that there is a novel family of matrix proteins that may use immunoglobulin motifs as binding sites for cellular integrins and that there are matrix proteins in addition to AAAP-40 that may serve as autoantigens in the pathogenesis of AAA disease. PMID- 9224394 TI - Nitric oxide contributes to adriamycin's antitumor effect. AB - Recently, several antitumor drugs have been shown to stimulate nitric oxide (NO) production. PURPOSE: To determine if adriamycin induces NO production in breast cancer cells in vitro and whether NO contributes to adriamycin's antitumor effect in vivo. METHODS: Murine breast cancer cells (EMT-6) were incubated with adriamycin (ADRIA, 0, 10, 100, 1000 microM) in the presence or absence of the NO synthase inhibitor aminoguanidine (AG, 1 mM). Twenty-four hours later nitrite accumulation (Greiss reagent) and cell viability (MTT assay) were assessed. Supernatants from adriamycin-stimulated cells were also analyzed at 6, 8, and 24 hr for TNF, IL-1, and IFN gamma (ELISA). For in vivo experiments, 10(5) EMT-6 cells were injected into the flank of BALB/c mice (n = 20) and 1 hr later mice received one of four treatments: (1) saline, (2) ADRIA (10 mg/kg ip), (3) AG (100 mg/kg sc BID), or (4) ADRIA (10 mg/kg ip) and AG (100 mg/kg sc BID). Two weeks later tumor size was measured and in situ tumor cell apoptosis was determined by fluorescent microscopy and flow cytometry. RESULTS: Adriamycin was cytotoxic to EMT-6 cells with 100 microM resulting in nearly 100% killing (P < 0.01). Adriamycin also stimulated nitrite accumulation with 100 microM producing 6.5 +/- 0.26 microM nitrite (P < 0.001). AG blocked adriamycin-stimulated nitrite accumulation (P < 0.05), but did not inhibit cytotoxicity in vitro. In vivo, adriamycin inhibited tumor size by nearly 400% (P < 0.001), while AG attenuated adriamycin's effect on tumor growth (P < 0.05). There was no difference in the detection of apoptotic tumor cells between the adriamycin and adriamycin and AG groups as determined by immunohistochemistry and flow cytometry. CONCLUSIONS: These findings suggest that adriamycin stimulated NO production in EMT-6 cells, but adriamycin's cytotoxicity in vitro was NO-independent. In vivo, adriamycin inhibited tumorigenesis partially via an NO-dependent, nonapoptotic mechanism. PMID- 9224395 TI - Hemodynamic and inotropic effects of nitric oxide in pulmonary hypertension. AB - Right ventricular failure following cardiac transplantation is most commonly related to pre-existent recipient pulmonary hypertension secondary to chronic congestive heart failure. Although nitric oxide has had some role clinically in improving pulmonary hemodynamics and right ventricular function in this setting, an appropriate large-animal model of stable pulmonary hypertension has not been available for basic investigation of this problem. This study was designed to examine the hemodynamic and inotropic effects of inhaled nitric oxide using a canine model of monocrotaline pyrrole-induced chronic pulmonary hypertension. Eight mongrel dogs (22-25 kg) were used. All animals underwent percutaneous pulmonary artery catheterization to measure right heart hemodynamics prior to and 8 weeks after a right atrial injection of monocrotaline pyrrole. Eight weeks post injection, all hearts were instrumented with a pulmonary artery flow probe, sonomicrometric dimension transducers, and micromanometers. Data were collected at baseline and following nitric oxide administration. Eight weeks post monocrotaline pyrrole injection, significant increases were observed in the pulmonary hemodynamics compared to pre-injection. Nitric oxide led to significant decreases in pulmonary vascular impedance. Significant improvements in pulmonary blood flow, transpulmonary efficiency, and right ventricular contractility were also observed. This investigation demonstrates the well-known clinical effects of nitric oxide in improving pulmonary hemodynamics which were also associated with an increase in pulmonary blood flow, transpulmonary efficiency, and right ventricular contractility in the setting of monocrotaline pyrrole-induced pulmonary hypertension. PMID- 9224396 TI - Nifedipine protects small intestine from cyclosporine-induced hemodynamic and functional impairment. AB - We have previously shown that cyclosporine (CsA) causes intestinal hemodynamic and functional impairments. In this study, we evaluated whether nifedipine protects the small intestine from such toxic side effects. Isogeneic small intestinal transplantation was performed in rats which then received one of the following two-week treatments: olive oil, 0.15 ml/kg/day i.m. as vehicle controls in group 1; nifedipine, 1 mg/kg/day i.m. in group 2; CsA, 15 mg/kg/day i.m. in group 3; and both nifedipine and CsA in group 4. Vascular resistance, whole tissue blood flow and its mucosal and serosal/muscularis distributions in both graft and recipient residual native intestines, and absorptive function were determined. The data showed that two-week treatment with CsA resulted in a marked elevation of vascular resistance from 51.0 +/- 6.8 to 72.4 +/- 11.1 U/g in the native whole tissue and from 53.7 +/- 7.2 to 78.2 +/- 12.1 U/g in the graft whole tissue, and decreases in blood flow from 1.59 +/- 0.26 to 1.11 +/- 0.17 ml/g/min in the native whole tissue and from 1.50 +/- 0.21 to 1.03 +/- 0.18 ml/g/min in the graft whole tissue and absorption from 227 +/- 36 to 166 +/- 26 mg glucose/dl. Mucosa was preferentially affected, while serosal/muscularis layers remained relatively unchanged. When nifedipine was concomitantly used with CsA, vascular resistance and blood flow values in the mucosal layer and whole intestinal tissue as well as absorptive function showed no significant differences from the baseline data. The changes observed in denervated grafts and recipient native intestines were similar. We conclude that nifedipine is effective in protecting both graft and native small intestines from CsA-induced toxicity in the rat. PMID- 9224397 TI - Boundary layer infusion of basic fibroblast growth factor accelerates intimal hyperplasia in endarterectomized canine artery. AB - We examined the effects of human recombinant basic fibroblast growth factor (bFGF) on the proliferation and migration of cultured dog smooth muscle cells (SMCs) and endothelial cells (ECs) and the effect of continuous local boundary layer infusion of bFGF on intimal hyperplasia in endarterectomized dog artery. In vitro proliferation and migration of dog SMCs or ECs were performed using direct counting and Boyden's chamber, respectively. At a dose of 10 ng/mL, bFGF significantly promoted both SMC and EC proliferation (7- and 4-fold, respectively) and migration (2.3- and 1.9-fold, respectively). Six dogs underwent bilateral carotid endarterectomies. A newly designed local infusion device with an osmotic pump continuously delivered bFGF to one artery or vehicle solution to the contralateral artery for 14 days. The intimal thickness and area in the bFGF treated vessels were increased by 72 and 81%, respectively, compared with control arteries (P < 0.05). As assessed by the bromodeoxyuridine index, the proliferative activity was increased by 73% in bFGF-treated arteries (P = 0.03). Furthermore, cell proliferation at the distal anastomoses of local infusion device was significantly increased in the bFGF-infused grafts compared with distal anastomoses in the control grafts (13.24 +/- 1.24% versus 5.24 +/- 1.01%, P < 0.01). These data demonstrate that human recombinant bFGF has a potent effect on dog SMC and EC proliferation and migration, and that local infusion of exogenous bFGF significantly enhances the intimal hyperplasia formation and cell proliferation to vascular injury. We conclude that the bFGF pathway may contribute to the development of intimal hyperplastic lesions. PMID- 9224398 TI - Graft versus host disease in rats made tolerant for organ allografts. AB - Tolerance for organ allografts would eliminate acute and chronic rejection as well as the need for nonspecific immunosuppression. A potential hazard of tolerance is the susceptibility to graft vs host disease (GVHD) due to unresponsiveness to alloantigen. This study sought to determine if our model of tolerance induction results in susceptibility to GVHD. Chimeras were created by transplantation of T-cell depleted ACI and Lewis bone marrow into lethally irradiated Lewis rats. Chimerism was determined post-BMTx by flow cytometric analysis of recipient spleens for the presence of ACI cells. ACI/Lew chimeras (ALC), animals that reconstituted only with syngeneic (Lewis) marrow (so-called failed chimeras), and ACI/Lew F1 (LACF1) hybrid rats were all given 200 x 10(6) ACI splenocytes i.v. Animals were examined for evidence of GVHD. GVHD was quantified using the popliteal lymph node enlargement assay. All LACF1 (n = 6) rats developed severe lethal GVHD following ACI splenocyte injection. Similarly, ALC (n = 6) developed fatal GVHD. Animals that reconstituted only with syngeneic Lewis marrow (failed chimeras) showed no signs of illness. GVHD was confirmed histologically and immunohistochemically. Failed chimeras receiving ACI splenocyte challenge showed no evidence of GVHD histologically. Popliteal lymph node enlargement indices reflected the presence of GVHD in the chimeras and hybrids but not in the failed chimeras. We conclude that tolerance induction by mixed chimerism results in susceptibility to GVHD if enough donor lymphoid tissue is given to the host at the time of organ transplant. Animals that are not mixed chimeras (failed bone marrow transplant) rejected the allogeneic splenocytes as evidenced by their lack of disease. Tolerance may therefore make the host defenseless against fatal GVHD. PMID- 9224399 TI - Decreased expression of LFA-1 on peripheral blood lymphocytes in graft versus host disease. AB - We have previously demonstrated an increase in lymphocyte function associated antigen-1 (LFA-1) expression in the native intestines of animals with graft versus host disease (GVHD) after small bowel transplantation (SBTx). The present study evaluated LFA-1 expression on peripheral blood lymphocytes (PBLs) during GVHD. GVHD was created in LBNF1 rats by heterotopic vascularized SBTx from Lewis donors and compared to sham-op controls and cyclosporine-treated SBTx rats (SBTx CyA, 10 mg/kg/day). PBLs were harvested on Postop Day 13 when signs of severe GVHD were present and PBLs were stained for LFA-1, CD3 (T-cell marker), and CD45 (B cell marker) and analyzed on an Epics 5 flow cytometer. Mesenteric lymph node (MLN) lymphocytes from the native intestines were also harvested for each group and stained for LFA-1. There were significant decreases in LFA-1, CD3, and CD45 PBL expression in rats with GVHD. CyA treatment corrected the abnormal CD3 and CD45 ratios, but not LFA-1 expression. It is concluded that: (1) The proportion of PBLs expressing LFA-1 is depressed in animals with clinical GVHD consistent with their recruitment into the host's inflamed tissues. (2) CyA treatment corrects the abnormalities in T and B cell ratios but not the decreased expression of LFA-1 on PBLs and may relate to its known imperfect ability to treat GVHD. PMID- 9224400 TI - Chronic hepatitis C virus infection in humans: induction of hepatic nitric oxide synthase and proposed mechanisms for carcinogenesis. AB - Chronic inflammatory states frequently lead to the increased production of nitric oxide (NO) via inducible NO synthase (NOS-2). In addition, NO may produce mutagenesis through several mechanisms such as DNA oxidation, DNA deamination, and the formation of N-nitroso compounds. As there is a strong association between human hepatitis C virus (HCV) infection and the development of hepatocellular carcinoma (HCC), we were interested in whether human HCV hepatitis leads to induction of NOS-2 and if the mutation repair system of p53/p21 was upregulated. Reverse transcriptase-polymerase chain reaction (RT-PCR) for human NOS-2 message was performed on RNA samples from both liver biopsies and whole liver from HCV-positive and control patients (normal liver from hepatic resections for metastases). Immunohistochemistry (IHC) for p53 and Western blot analysis for p21 were also performed on the whole liver samples. From the liver biopsies, 60% of HCV-positive patients expressed NOS-2 by RT-PCR. Looking at the whole liver samples, 100% of the HCV-positive patients expressed NOS-2 vs 12.5% in the normal samples. p53 was not detected in either group but there was upregulation of p21 over baseline expression in a number of the HCV-positive patients. Human HCV hepatitis leads to consistent upregulation of hepatic NOS-2 message, but message is not predictably present in "normal" human liver. There is also induction of p21 in some patients with HCV hepatitis. Chronic expression of NO in HCV hepatitis may play a role in DNA mutagenesis and the development of HCC. PMID- 9224401 TI - Starvation enhances hepatic free radical release following endotoxemia. AB - Although it is well known that malnourished patients who become septic have an increased risk of organ failure and death compared to normally nourished individuals, the pathological processe(s) underlying this observation are unknown. To evaluate one possible explanation for this finding, we tested the hypothesis that malnutrition depresses hepatic antioxidant stores and accelerates hepatic release of oxygen free radicals in an animal model of sepsis. Male rats were either fasted (n = 14) or fed (n = 14) for 3 days prior to receiving lipopolysaccharide (LPS, 17 mg/kg intraperitoneally). Animals were weighed daily and then sacrificed 6 and 24 hr after LPS administration to determine hepatic superoxide anion (an oxygen free radical) release and liver glutathione (GSH, an antioxidant) content. Fasted rats were severely malnourished as indicated by a 23% decrease in body weight compared to fed rats, which gained 11% (P < 0.05). Liver GSH was depressed by 30% (P < 0.05) and 20% (P = 0.066) in the fasted compared to fed animals 6 and 24 hr after LPS administration. In addition, hepatic superoxide anion release was 210 and 75% higher in the fasted animals 6 and 24 hr after LPS injection (P < 0.05 at both time points). Liver superoxide anion release and GSH content were negatively correlated (P < 0.001, R = - 0.73) indicating that superoxide anion release increased as GSH content fell. Malnutrition leads to depletion of liver antioxidant stores with accelerated release of hepatic oxygen free radicals. Oxidant-mediated organ damage may be one cause of increased morbidity and mortality in malnourished, systemically infected patients. PMID- 9224402 TI - Dexamethasone and lipopolysaccharide regulation of taurine transport in Caco-2 cells. AB - Intracellular enterocytic levels of the immunomodulator taurine decrease significantly in response to trauma and surgical insult. The effect of physiological stress on enterocyte taurine uptake is unknown. The aim of this study was to compare taurine transport under basal and stressed conditions using the human intestinal Caco-2 cell line in vitro. Caco-2 cells were incubated with 10 nM [1,2-3H]taurine at 37 degrees C and 5% CO2 and taurine uptake was examined over the range of 0.1-10 microM to determine kinetic parameters of the transporter. The culture medium was then supplemented with dexamethasone and/or lipopolysaccharide (LPS) and taurine uptake was calculated as picomoles per milligram protein per hour. Statistics were by unpaired Student's t test. Taurine uptake was hyperbolically related to taurine concentration and obeyed Michaelis Menten kinetics with a K(m) of 5.27 +/- 0.95 microM and Vmax of 1125.43 +/- 130.9 pmole/mg protein/ hour. Dexamethasone (1-1000 microM) significantly reduced taurine uptake by up to 66.15%. LPS (1 microgram/ml) impaired transport of taurine by 15.7%, and in combination with dexamethasone (100 microM) by 42.4%. All results are mean of at least three experiments and P < 0.05. We have established that taurine uptake by enterocytes is downregulated by dexamethasone. This may relate to the decreased intestinal levels of taurine observed in trauma and surgery patients. Further study may elucidate mechanisms whereby homeostasis of enterocyte taurine might be maintained during sepsis. PMID- 9224403 TI - The use of transgenic mice to generate high affinity p53 specific cytolytic T cells. AB - P53 is an attractive target immunotherapy because it is overexpressed in up to one half of all malignancies, and its overexpression often correlates with a worsened prognosis. We wanted to determine the feasibility of targeting wild-type epitopes p53 on human tumor cells. HLA A2.1 transgenic mice were immunized with the immunodominant wild-type p53 peptide epitopes, p53(149-157) and p53(264-272), along with a pan-DR helper epitope peptide in incomplete Freund's adjuvant (IFA). Twelve days later, splenocytes were harvested and stimulated with syngeneic blast cells that had been acid-treated to remove endogenous peptide and p53 peptide pulsed. The responding cells were subsequently restimulated weekly with acid washed, peptide-pulsed Jurkat cells transfected with HLA A2.1. Peptide specific activity was tested in a chromium release assay. The resulting cytotoxic T cells (CTL) were cloned by limiting dilution. Peptide specific CTL were generated against both p53(149-157) and p53(264-272. Only p53(149-157) specific CTL were able to recognize and lyse cells that overexpressed endogenous p53. CTL clones derived from the p53(149-157) cell line demonstrated high affinity and specificity for p53(149-157) when presented by HLA A2.1+ cells. The p53(149-157) specific CTL were tested for specificity against a variety of cultured human cell lines. The CTL clones only lysed cells that overexpressed p53 in the context of HLA A2.1 and did not lyse cells with normal p53 expression or cells that lacked HLA A2.1 expression. This study demonstrates the possibility of targeting tumors, which overexpress p53, and raises the possibility transferring the high affinity, p53 specific T cell receptors from the murine CTL to human T cells. PMID- 9224404 TI - Deoxycholate inhibits in vivo butyrate-mediated BrDU labeling of the colonic crypt. AB - The short-chain fatty acid butyrate (NaBu) selectively increases colonic crypt base proliferation and inhibits "premalignant" crypt surface hyperproliferation while the secondary bile acid deoxycholate (DCA) induces surface hyperproliferation, in vitro. We hypothesized that NaBu and DCA have similar selective and antagonistic effects on the colonic crypt proliferative pattern, in vivo. Fifty-six adult SD rats underwent surgical isolation of the colon and 24-hr intraluminal instillation with physiological (10 mM) and pharmacological (25 mM) levels of butyrate alone or combined with a physiological DCA level (5 microM). Bromodeoxyuridine-labeling indices (LI) were determined as labeled cells divided by total cells, for the whole crypt and five crypt compartments from base to surface. Treatment with NaBu increased total LI when compared to NaCl. This effect was significant only at the crypt base. Both doses of NaBu resulted in similar LI with no further response at the higher concentration. In contrast to prior in vitro studies, DCA alone at this concentration did not affect LI, but when combined with NaBu, DCA inhibited the effects of NaBu at the crypt base and surface. The conclusions are: (1) the in vivo proliferative effects of NaBu are selective to the crypt base, (2) an in vivo low physiological DCA level does not promote crypt surface hyperproliferation but does inhibit butyrate's proliferative effect, and (3) NaBu and DCA interact in a complex and antagonistic manner to selectively modulate crypt base and surface proliferation, in the rat colon, in vivo. These findings may have clinical relevance since colonic levels of NaBu and DCA are affected by diet. PMID- 9224405 TI - The temporal expression of transforming growth factor-beta 1 in early aortocoronary vein grafts. AB - The success of coronary reconstructive procedures is limited by the high incidence of restenosis secondary to intimal hyperplasia (IH). Transforming growth factor-beta 1 (TGF-beta 1) is a growth factor which has been shown to be important in the early development of IH in arteries and peripheral vein grafts. To date, there is little information concerning the early remodeling in aortocoronary vein grafts (ACVG). The purpose of this study was to characterize the expression of TGF-beta 1 expression in early aortocoronary vein grafts. Eighteen mongrel dogs underwent aortocoronary vein bypass grafting. Vein grafts were excised at 2 hr, 4 hr, and 7 days after implantation, snap frozen, and processed for ribonuclease protection assays (RPA) using 32P-labeled riboprobes for TGF-beta 1 and 18 S rRNA. TGF-beta 1 expression was quantified by densitometric analysis of autoradiographs which were expressed as a ratio TGF beta 1/rRNA. Representative vessel rings were also collected for histology. There was a significant rise in TGF-beta 1 expression in the 2-hr vein grafts (0.42 +/- 0.04 compared to control saphenous vein (0.21 +/- 0.05, P < 0.02). In addition, there was significant downregulation of TGF-beta 1 at 4 hr (0.28 +/- 0.05) and at 7 days (0.18 +/- 0.01) when compared to 2 hr (P < 0.05). Histological specimens showed minimal intimal hyperplasia at 7 days. These results show for the first time an acute rise in TGF-beta 1 expression in ACVG. This upregulation quickly subsides by 4 hr and gene expression approaches control values by 7 days. By understanding this temporal relationship of expression one could better target potential therapeutic modalities to attenuate IH. PMID- 9224407 TI - One-on-one mentor-resident rotation for improving continuity of care in a surgical training program. AB - The modern resident team, staffed by multiple attendings, often makes sacrifices on continuity of care due to scheduling conflicts. We investigated a one-on-one mentor-resident rotation where all clinical activities were synchronized to produce near-perfect continuity of care, and we compared the objective and subjective outcome measures to those of control rotations of the same resident during the same year. The results showed that continuity of care close to 100% was possible in such rotations, but work hours were increased by 25%. Also, the number of patients seen was decreased by over 50%. The rotation was well-received by both mentors and residents. Continuity of care per se can be improved by this rotation. However, theoretical disadvantages, mainly narrow training due to exposure to only one mentor and fewer patients, make it unsuitable for extended use. PMID- 9224406 TI - EGF and IGF-I synergistically stimulate proliferation of human esophageal epithelial cells. AB - Proliferation of esophageal mucosal cells is important in repair of reflux induced injury. We studied the effects of EGF, IGF-I, and IGF-I/binding protein-3 (BP-3) complex on immortalized esophageal epithelial (HET-1A) cells and searched for synergy between the growth factors. HET-1A cells were plated at 5 x 10(4) in 12 well plates. After 2 days in optimal media, they were maintained in basal media with or without the test peptides: EGF at 0.05-5 nM, IGF-I at 0.1-10 nM, IGF-I/BP-3 at 0.1-10 nM, and a combination of EGF at 5 nM and IGF-I or IGF-I/BP-3 at 1 and 10 nM. In comparison to basal media EGF and IGF-I stimulated cell proliferation over baseline at 5 and 10 nM, respectively. The combination of EGF at 5 nM and IGF-I at 1 and 10 nM worked synergistically, increasing cell counts over baseline to 10.3 +/- 0.2 and 14.6 +/- 0.8 x 10(5), respectively. The calculated additive effect of EGF and IGF-I at 1 and 10 nM individually increased cell counts to 8.2 +/- 0.2 and 10.4 +/- 0.6 x 10(5), respectively. The difference between the observed and the calculated values was significant at P < 0.05, ANOVA, Turkey test. IGF-I/BP-3 complex enhanced this synergy at low levels of IGF I but not at 10 nM IGF-I. EGF, IGF, and IGF-I/BP-3 independently promote HET-1A proliferation. IGF and EGF in combination demonstrate synergism with potentiated interaction presumably because of their different roles in the cell cycle, EGF being a competence factor and IGF being a progression factor. This combination may have potential as a treatment for esophageal mucosal injury, and IGF-I/BP-3 may further enhance their benefit. PMID- 9224408 TI - Increased nitric oxide in exhaled gas is an early marker of hypovolemic states. AB - Acute hemorrhage is associated with a variety of physiologic and metabolic alterations, including vascular hyporeactivity and endothelial cell dysfunction. The lung is a major target organ during hemorrhagic shock. The effect of acute hemorrhage on NO production in the lung is not well described. In the present study we examined the effect of acute hemorrhage on exhaled NO (NOe), and studied how changes in blood volume and flow affect NOe. Anesthetized and mechanically ventilated rabbits were used. The effect of acute hemorrhage by slow exsanguination on NOe was examined using chemiluminescence. Because hemorrhagic shock is associated with decreased pulmonary blood flow, we established an isolated lung preparation perfused with autologous blood (Hct = 17.4%) and studied the effect of pulmonary flow rate on NOe independent of metabolic changes. In order to separate the effect of flow from the effect of changes in blood volume, we examined the effect of flow in isolated lungs perfused with a blood-free albumin solution (PAS). In the isolated lung, ventilation was similar to that used in the intact animal, and temperature, pH, pCO2, and PO2 were kept normal. Prior to exsanguination, baseline NOe in the intact animal was 24 +/- 3 ppb. At 5, 10, 15, and 20 min after initiating the hemorrhage, NOe rose to 31 +/- 3, 51 +/- 7, 94 +/- 10, and 154 +/- 16 ppb, respectively (P < 0.05). During baseline conditions in the blood-perfused isolated lungs (200 ml/min), NOe was 35 +/- 3 ppb. When flow was decreased to 70 and 0 ml/min, NOe increased to 37 +/- 3 and 56 +/- 6 ppb, respectively (P < 0.001). During baseline conditions in the PAS perfused lungs (70 ml/min), NOe was 94 +/- 13 ppb and was unaffected by changes in flow. The perfusion pressure in the isolated lungs was in the normal range. Reduction in blood flow rate in the isolated lung was associated with less than twofold increase in NOe. This was attributed to reduction in red blood cell volume and not due to changes in blood flow rate. Reduction in flow in the intact animal during hemorrhage generated more than threefold increase in NOe, suggesting that neurohumoral mediators, in addition to changes in flow, play an important role in determining. NOe in the intact condition. NOe began to rise immediately after exsanguination began, and therefore may be a useful early marker of acute hemorrhagic shock and hypovolemia. This information may be useful in the intensive care setting. PMID- 9224409 TI - Matrix metalloproteinase inhibition attenuates human pancreatic cancer growth in vitro and decreases mortality and tumorigenesis in vivo. AB - Matrix metalloproteinases (MMP) are enzymes responsible for extracellular matrix degradation, a critical component influencing the growth and metastatic potential of cancer. The purpose of this study was to determine the in vitro effects of MMP inhibition on human pancreatic cancer cells and to document its effect on cancer growth in vivo. The effect of MMP inhibition was determined using the MMP inhibitor BB-94 and a moderately differentiated pancreatic cancer cell line (HPAC). In vitro, a dose response curve was generated over 5 days utilizing the MTT [3(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. In vivo, using an established orthotopic model for pancreatic cancer (LD100 = 80 days), 22 nude mice with orthotopic tumors (30 were implanted) received either BB-94 or vehicle beginning 4 days prior to implantation and continuing to death or sacrifice on Day 70. Mice were weighted weekly. At death/sacrifice, tumors were weighted, volume determined, and metastases/ distant spread documented. In vitro, BB-94 had little effect on HPAC proliferation at 40 ng/ml but achieved progressively greater to near complete inhibition at doses up to 4000 ng/ml while maintaining cell viability. In vivo, BB-94 significantly increased length of survival (69 +/- 0.1 days vs. 56 +/- 3.1 days) and necropsy weight (25.7 +/- 1.67 g vs. 19.8 +/- 1.14 g) while decreasing metastatic rate (1 vs. 20) and tumor size (0.14 +/- 0.02 g vs. 0.65 +/- 0.1 g). MMP inhibition limits HPAC proliferation in a dose-dependent fashion without direct cytotoxic effects in vitro. Mice harboring orthotopic tumors treated with BB-94 demonstrated significant reductions in tumor weight, volume, and metastases which corresponded to increased animal weight and prolonged survival. PMID- 9224410 TI - Prostaglandin E2 modulates monocyte MHC-II (Ia) suppression in biomaterial infection. AB - Staphylococcus epidermidis biomaterial infection is associated with local cellular immune suppression measured by a depressed monocyte (M phi) Ia expression. The purpose of this study was to define the effect of proinflammatory mediators on Ia expression and bacterial clearance in experimental biomaterial infection. A 1-cm-long Dacron tube graft, sterile or colonized with Staphylococcus epidermidis (1 x 10(7) cfu/ml2), was implanted in Swiss-Webster mice. Perigraft fluid was collected at 7, 10, 14, and 28 days and assayed by enzyme-linked immunoassays for tumor necrosis factor alpha (TNF alpha), interleukin (IL)-I alpha, IL-4, IL-10, and prostaglandin E2 (PGE2). Grafts were sonicated and plated for quantitative growth. In vivo effector inhibitions was accomplished with anti-TNF alpha, anti-IL-1 alpha antibodies (7 micrograms/24 hr), or indomethacin (50 micrograms/24 hr) via an Alzet 7-day microinfusion pump. M phi Ia expression was determined by flow cytometry. A significant elevation of TNF alpha, IL-1 alpha, and PGE2 was found during the first 10 days in the infected compared with sterile (P < or = 0.05) grafts and correlated with maximal Ia suppression. Neither IL-4 nor IL-10 was significantly different in the sterile or infected perigraft fluid. Indomethacin completely prevented M phi Ia suppression, while anti-IL-1 alpha only partially (94%) prevented M phi Ia suppression with a corresponding decrease in PGE2 production in infected grafts. Anti-TNF alpha increased PGE2 production by 189% and was associated with depressed M phi Ia expression. Indomethacin treatment improved mean graft adherent bacterial clearance by 54% at 7 days and 75% at 28 days compared with control (not significant). Interleukin-1 alpha but not TNF alpha increases PGE2 production which modulates M phi Ia suppression. To improve treatment of biomaterial infections, local immunomodulation of PGE2 and IL-1 alpha is promising. PMID- 9224411 TI - Alterations in oxidative metabolism and glutamine transport support glucose production in the tumor-influenced hepatocyte. AB - Glutamine is the primary substrate whose hepatic transport is upregulated in the tumor-bearing host; however, the subsequent metabolism of transported glutamine is currently unknown. The purpose of this study was to determine if glutamine is an important oxidative fuel source for hepatocytes in cancer. Specifically we compare rates of glutamine transport and oxidation in hepatocytes from control and tumor-bearing animals. We also compare rates of glucose oxidation and rates of glucose production from glutamine in control hepatocytes versus those from tumor-bearing animals. Hepatocytes from rats bearing the MCA fibrosarcoma were isolated when tumors comprised 5 and 15% of total body weight and compared to sham-implanted and pair-fed control animals. [3H]GLN transport, GLN and glucose oxidation to CO2, and glucose production from glutamine were assayed. Tumor burden of 5% stimulated a 2.52-fold increase in hepatocyte glutamine transport and a 2-fold increase when tumor burden reached 15%. Rates of oxidation of glutamine were suppressed by 1.5-fold when tumors comprised 5% of body weight compared to sham animals and were equivalent to sham animals when tumors comprised 15% of body weight. Significant alterations in glucose oxidation were observed when tumors were both small and large-glucose oxidation was suppressed by 3.6- and 3.7-fold when tumors comprised 5 and 15% of body weight respectively compared to sham-implanted rats. Incubation of hepatocytes from tumor-bearing animals with glutamine as a gluconeogenic substrate induced a 1.84-fold increase in glucose production compared to sham hepatocytes. In conclusion, (i) despite a doubling of GLN transport by the tumor-influenced hepatocyte, GLN oxidation by hepatocytes was not increased. (ii) Glucose oxidation by hepatocytes from tumor bearing animals was decreased compared to sham hepatocytes and, simultaneously, glucose production by tumor-influenced hepatocytes from glutamine was increased. The augmentation of hepatic glutamine transport and decreased glutamine oxidation seen in tumor-influenced hepatocytes appear to support hepatocyte gluconeogenesis from glutamine. PMID- 9224412 TI - Reduced ischemia-reperfusion injury with isoproterenol in non-heart-beating donor lungs. AB - Transplantation of lungs retrieved from non-heart-beating donors could expand the donor pool. Recent studies suggest that the ischemia-reperfusion injury (IRI) to the lung can be attenuated by increasing intracellular cAMP concentrations. The purpose of this study was to determine the effect of IRI on capillary permeability, as measured by Kfc, in lungs retrieved from non-heart-beating donors and reperfused with or without isoproterenol (iso). Using an in situ isolated perfused lung model, lungs were retrieved from non-heart-beating donor rats ventilated with O2 or not at varying intervals after death. The lungs were reperfused with or without iso (10 microM). Kfc, lung viability, and pulmonary hemodynamics were measured, and tissue levels of adenine nucleotides and cAMP were measured by HPLC. Iso-reperfusion decreased Kfc significantly (P < 0.05) compared to non-iso-reperfused groups at all postmortem ischemic times, irrespective of preharvest ventilation status. Pulmonary arterial pressures and resistances increased and venous resistances decreased with iso-reperfusion. Total adenine nucleotide (TAN) levels correlated with Kfc in non-iso-reperfused (r = 0.65) and iso-perfused (r = 0.84) lungs. cAMP levels increased significantly with iso-reperfusion. cAMP levels correlated with Kfc (r = 0.87) in iso reperfused lungs. Iso-reperfusion of lungs retrieved from non-heart-beating donor rats results in decreased capillary permeability and increased lung tissue cAMP levels. Pharmacologic augmentation of tissue TAN and cAMP levels may further ameliorate the increased capillary permeability seen in lungs retrieved from non heart-beating donors. PMID- 9224413 TI - Pravastatin increases survival and inhibits natural killer cell enhancement factor in liver transplanted rats. AB - Pravastatin, a 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitor, has been shown to decrease the number of acute rejection episodes in cardiac and renal transplant patients. This study evaluates the effects of pravastatin on survival of rats following liver transplant and attempts to elucidate the mechanisms of these effects. Both survival and natural killer cell enhancing factor (NKEF) studies utilized Dark Agouti rats for donor livers transplanted into Brown Norway rats as recipients. All rats received daily low-dose cyclosporine (CsA) 2 mg/kg/day by gavage. The treated groups also received gavage doses of pravastatin, 20 mg/kg/day. Survival data were analyzed by the method of Kaplan-Meier and log-rank chi 2 tests for statistical significance. For NKEF evaluation, rats were sacrificed at varying time points; total RNA was extracted from the liver and hybridized with 32P-radiolabeled NKEF DNA probes in the Northern blot technique. Radiographs were quantitated using densitometry. Data were analyzed by two-way ANOVA. Actuarial survival was improved (P < < 0.05) in rats treated with pravastatin in addition to low-dose CsA (n = 41, CsA alone n = 74). Less fibrosis and chronic rejection was seen on histological section in the treated animal livers, P < 0.05, NKEF was seen maximally at Days 5-15 tapering off at Day 21. NKEF-a and NKEF-b levels were significantly decreased in the animals treated with CsA and pravastatin compared to CsA alone in the group of animals < 16 days postop (P < < 0.05). Pravastatin improves survival in rats following OLT and while the mechanism is still unknown, inhibition of natural killer cell enhancement factor may represent an alteration in the overall immune response. PMID- 9224414 TI - Candida infection following severe trauma exacerbates Th2 cytokines and increases mortality. AB - Following trauma, there is an increase of Th2 cytokines (IL-4, IL-6, and IL-10) and a decrease in Th1 cytokines (IFN-gamma and IL-2) that may account for impaired cellular immunity. However, the functional significance of a dominant Th2 pattern to the host remains unclear. The aim of this study was to evaluate whether Candida albicans (CA) sepsis in the setting of a Th2 response to trauma leads to increased mortality and to examine the mediators involved. Female BALB/c mice were randomized (12 per group) to receive no injury (C); trauma, consisting of a combined femur fracture and 40% total blood loss (T); no injury plus CA infection (C+CA); and CA infection 1 week following trauma (T+CA). Survival was then followed for 3 weeks. In a separate study, mice were treated as above (5 per group) and sacrificed. Harvested splenocytes were evaluated for concanavalin A stimulated cytokine production and liver and kidney homogenates were plated to evaluate CA growth per organ and examined histologically. Candida infection at 1 week following trauma resulted in significantly increased mortality compared to infected controls. Furthermore, the Th2 dominant cytokine pattern was significantly augmented in the presence of CA infection in both C+CA and T+CA groups. Additional analysis showed significant growth of CA in liver and kidney homogenates from T+CA compared to C+CA mice. These results suggest that injured and infected mice demonstrate augmentation of Th2 dominant responses above that of injury or infection alone, as well as a decreased ability to clear Candida which may partially explain the increase in mortality observed. Therapies designed to neutralize Th2 cytokines or augment Th1 cytokines may prove beneficial in the setting of sepsis following trauma. PMID- 9224415 TI - Macrophage TNF mRNA expression is modulated by protease inhibitors. AB - BACKGROUND: Nuclear factor kappa B (NF kappa B) is an important transcriptional activator protein and is a crucial component of the host's response to infection. The activation of NF kappa B is correlated with the phosphorylation of inhibitory kappa B (I kappa B) and its subsequent degradation. We hypothesized that protease inhibitors which prevented I kappa B degradation could inhibit the macrophage gene activation and reduce the production of inflammatory cytokines. METHODS: Rabbit alveolar macrophages (M phi) were obtained by bronchoalveolar lavage. M phi were exposed to Escherichia coli lipopolysaccharide (LPS) (10 ng/ml) in the presence of various concentrations of protease inhibitors, either N-tosyl-L phenylalanine chloromethyl ketone (TPCK) or N-benzoyl-L-tyrosine ethyl ester (BTEE). Total RNA was extracted for Northern blot assay of tumor necrosis factor (TNF) mRNA expression using a rabbit genomic DNA probe. Total nuclear extracts were also obtained for the measurement of the NF kappa B activity with the electrophoretic mobility shift assay. The TNF production in the M phi supernatant was measured by L929 bio-assays. RESULTS: NF kappa B activity induced by LPS was inhibited by either BTEE or TPCK. Inhibition of NF kappa B activity by these agents also prevented TNF mRNA expression and TNF production induced by LPS. The cellular mechanism leading to NF kappa B activation was further studied. TNF mRNA expression and NF kappa B activation were inhibited by D609, a phospholipase C (PLC) inhibitor, as well as by protein kinase C (PKC) inhibitors. In addition, direct stimulation of PKC led to NF kappa B activation and TNF mRNA expression. CONCLUSIONS: These data suggest that TNF mRNA expression of LPS-stimulated M phi is mediated through NF kappa B, NF kappa B activation is intimately regulated by the PLC signaling pathway. PMID- 9224417 TI - Characterization of glutamine and glutamate transport in rat lung plasma membrane vesicles. AB - Insufficient glutamine for the lungs during sepsis may contribute to an impairment in lung function. Lung glutamine metabolism is supported by both blood glutamine uptake and de novo biosynthesis using circulating glutamate as a precursor. Information regarding the specific plasma membrane carriers involved in this uptake is lacking. Furthermore, the effect of sepsis on amino acid transport in whole lung has not been studied. We isolated lung plasma membrane vesicles (LPMVs) from control and LPS-treated rats and assayed glutamine and glutamate transport activity in LPMVs. Vesicle purity and functionality were confirmed by time-dependent concentrative amino acid uptake in the presence of Na+, impoverishment of microsomal enzymes, and a 25-fold enrichment in the plasma membrane marker 5'-nucleotidase. Eighty percent of glutamine uptake in lung vesicles was mediated via the high affinity Na(+)-dependent carrier System ASC (Vmax = 80 +/- 10 pmole/mg protein/15 sec; Km = 224 +/- 30 microM) while 19% occurred via the Na(+)-independent System ASC (Vmax = 11 +/- 2 pmole/mg/15 sec; Km = 141 +/- 23 microM). Ninety percent of glutamate transport was mediated by the Na(+)-independent System XAG-. Treatment of rats with LPS resulted in a decrease in both glutamine and glutamate transport in LPMVs. LPMVs offer a novel method for characterizing lung amino acid transport and studying the effects of catabolic states on this activity. The effects of endotoxin on System ASC and XAG activity may contribute to reduced lung glutamine availability during septic states which may impair cellular metabolism and function. PMID- 9224416 TI - The effect of ganciclovir on herpes simplex virus-mediated oncolysis. AB - Entry of herpes simplex virus (HSV) into tumor cells results in viral gene expression followed by cellular lysis. Attenuated HSVs selectively destroy tumors with sparing of surrounding normal tissue. HSV encodes a thymidine kinase (TK) that converts ganciclovir to a toxic metabolite. This metabolite may be transferred between cells and lead to the death of neighboring uninfected cells, termed bystanders. We sought to determine if HSV-mediated oncolysis is enhanced by ganciclovir treatment. In addition, we examined bystander killing in cocultures of TK transfectants and parental cells. hrR3, an attenuated HSV, expresses TK. The 50% lethal dose of hrR3 for a rat gliosarcoma (9L) and three human colorectal carcinomas (HT29, KM12C6, and KM12L4) was determined. Cells were infected with a 50% lethal dose of hrR3, followed by treatment with ganciclovir, and then cell survival was quantitated. In separate experiments 9L and HT29 cells were transfected with TK. Parental cells and TK transfectants were cocultured in various ratios, in the presence of ganciclovir, and cell survival was quantitated. hrR3-mediated oncolysis was enhanced by ganciclovir in the gliosarcoma but not in the three colorectal carcinomas. Cocultures of both 9L and HT29 parental cells with their corresponding TK transfectants demonstrated bystander killing. The mortality of 9L cocultures was 54% greater than that predicted for exclusive killing of transfectants. HT29 mortality was 8% greater than predicted. The ability of ganciclovir to augment hrR3-mediated oncolysis varies significantly between tumor cells lines. The extent of ganciclovir mediated killing of neighboring nontransduced parental cells similarly varies. Consideration should be given to these factors in the design of gene therapy strategies using HSV vectors as oncolytic agents. PMID- 9224418 TI - Exogenous human recombinant interleukin-10 attenuates hindlimb ischemia reperfusion injury. AB - Proinflammatory cytokines have been found to mediate part of the local and distant organ injury after ischemia and reperfusion (I/R). The anti-inflammatory cytokine interleukin-10 (IL-10) inhibits both TNF-alpha and IL-1, and we hypothesized that exogenous human IL-10 may decrease lung and soleus muscle injury after hindlimb I/R. Male Sprague-Dawley rats were randomly assigned to I/R (n = 10); I/R+IL-10 (10 micrograms i.v., n = 10), SHAM (n = 4); or SHAM+IL-10 (10 micrograms i.v., n = 4). Bilateral hindlimb ischemia was produced by tourniquet occlusion for 4 hr and all animals were sacrificed after 4 hr of reperfusion or at comparable times for the SHAMs. Soleus muscle cellular injury was determined by uptake of 99Tc pyrophosphate while soleus muscle capillary permeability, and lung capillary permeability were assessed by uptake of 125I-labeled albumin. Soleus muscle and lung neutrophil infiltration were measured with the myeloperoxidase assay. Serum samples were assessed for TNF-alpha production with the WEHI bioassay. Hindlimb I/R caused significant soleus muscle cellular injury, soleus muscle capillary injury, lung capillary injury, and lung neutrophil infiltration, Pretreatment with exogenous IL-10 significantly reduced soleus muscle capillary permeability and also reduced soleus muscle cellular injury, but not to a statistically significant degree. IL-10 administration also reduced pulmonary capillary permeability despite significantly increased lung neutrophil infiltration. Elevated TNF-alpha levels were found in 66% (4/6) rats in the I/R group versus 30% (3/10) rats in the I/R+IL-10 group. Exogenous IL-10 attenuates both local and distant organ injury after hindlimb I/R potentially independent of neutrophil infiltration. PMID- 9224420 TI - Induction of heat shock protein 72kDa expression is associated with attenuation of ischaemia-reperfusion induced microvascular injury. AB - Leukocyte-endothelial interaction is a pivotal step in the pathogenesis of ischemia-reperfusion (I/R) injury. Exposure of cells to a subcritical heat stress may protect cells from subsequent I/R injury, through induction of a 72-kDa heat shock protein (HSP72). The aim of this study was to investigate the effect of thermotolerance on leukocyte adherence and migration during an I/R period in rat mesenteric postcapillary venules. Sprague-Dawley rats were randomized into control (sham I/R), I/R, and thermotolerance+I/R groups. Thermotolerance was induced 18 hr prior to I/R, which was in turn established by occlusion of the superior mesenteric vascular pedicle for 10 mins, followed by 60 mins of reperfusion. The blood flow, leukocyte rolling velocity, and the number of adherent and migrated leukocytes in postcapillary venules were measured by intravital microscopy. I/R significantly decreased the rolling velocity of leukocytes; increased the number of adherent leukocytes at 10, 30, and 60 mins after reperfusion; and also increased the number of migrated leukocytes at 60 mins after reperfusion. Thermotolerance induction expression of HSP72 in pulmonary, intestinal, and mesenteric tissues was determined by Western immunoblotting. Thermotolerance significantly prevented the I/R-induced decrease in rolling velocity of leukocytes, the increase in the number of adherent leukocytes at 30 and 60 mins, and the increase in the number of migrated leukocytes at 60 mins. This results suggest that thermotolerance attenuates I/R injury by modulating leukocyte-endothelial interaction in vivo, possibly by increasing tissue expression of HSP72. PMID- 9224419 TI - Role of exogenous L-arginine in hepatic ischemia-reperfusion injury. AB - Plasma L-arginine is usually deficient immediately after hepatic reperfusion in orthotopic liver transplantation, which may also contribute to the occurrence of either hepatic ischemia-reperfusion injury or pulmonary hypertension. In this study, exogenous L-arginine was thus experimentally used to reverse the deficient status of the L-arginine/NO pathway. An in vivo model of 1 hr hepatic ischemia and reperfusion was thus tested in both rats (Experiment A) and pigs (Experiment B). In Experiment A, 10 mg/kg of L-arginine (group 1, n = 7), D-arginine (group 2, n = 7), or saline (group 3, n = 7) was administered through the portal vein. The hepatic tissue blood flow, at 20 min after reperfusion, improved in group 1 (70.7 +/- 7.0% of the preclamp levels) compared to groups 2 and 3. The serum glutamate oxaloacetate transaminase levels at 24 hr after reperfusion were also lower in group 1 (320 +/- 22.2 IU/L) than in either group 2 or group 3. The intrahepatic NO levels showed a temporal burst (> 15,000 pA current) after reperfusion only in group 1. In Experiment B, 10 mg/kg of L-arginine (group 4, n = 5), D-arginine (group 5, n = 5), or 10 ml of saline (group 6, n = 5) was administered through the portal vein. In group 4, the MPAP (mean pulmonary arterial pressure)/MAP (mean arterial pressure) was lower than that observed in groups 5 and 6. In conclusion, exogenous L-arginine administered from the portal vein was thus found to be effective in mitigating both portal hypertension and reperfusion injury by producing an increased amount of NO immediately after reperfusion. PMID- 9224421 TI - The response to calorie restriction in mammals shows features also common to hibernation: a cross-adaptation hypothesis. AB - The marked elevation in hepatic carbamyl phosphate synthetase I (CPSI) in calorie restricted mice, and the changes in erythrocyte 2,3-diphosphoglycerate (2,3-DPG) and in hemoglobin oxygen affinity in calorie-restricted and hypoxic humans living in Biosphere 2 suggest similarities between physiologic events in calorie restriction and hibernation. Other data from the literature strengthen this comparison. Accordingly, we hypothesize that the response to the calorie restriction regime as studied by gerontologists, rather than being a laboratory artifact, is part of a spectrum of responses to food deprivation which have adaptive value in the wild, and whose triggering mechanism may primarily involve the neuroendocrine system. PMID- 9224422 TI - Effect of life-long food restriction on cardiac myosin composition. AB - This study examined the effects of age and dietary manipulations on the cardiac myosin isozyme composition of male Fischer 344 rats. In hearts from ad libitum fed rats, aged 6-24 months, the myosin isozyme profile shifted with age from the fast, V1, to the slow, V3, isoform. Life-long food restriction (FR) (60% of ad libitum intake) as well as short-term FR (4 months) initiated at 16 months of age enhanced the age-related shift. Isocaloric reduction in the carbohydrate consumption of the rat from 2/3 to 1/3 of total calories had no effect on the isozyme profile, suggesting that FR acts via a decrease in calorie intake alone. The effect of FR on the cardiac myosin isozyme composition reported here shares several characteristics with the well-known effect of FR on life span extension, i.e., it (a) persists as long as FR is applied (life span extension is proportional to FR duration), (b) depends upon calorie reduction rather than a decrease in a specific dietary component, and (c) can be induced even when FR is initiated later in life. Suggesting that alterations in cardiac performance may be involved, the results may provide some clue as to the mechanism by which FR retards the aging processes. PMID- 9224423 TI - Adipose tissue-derived tumor necrosis factor-alpha activity is elevated in older rats. AB - High levels of adipose tissue-derived tumor necrosis factor-alpha (AT-TNF) mRNA and protein have previously been associated with genetic models of obesity and insulin resistance. Because there are endogenous TNF inhibitors it is unknown if AT-TNF activity is also increased. We hypothesized that AT-TNF activity would increase in older animals because of an accumulation of fat mass. We chose to study 2 different-aged male Fischer 344 rats, 3-month-old (young) and 14-month old (mature) because fat mass should be quite different but insulin action on glucose metabolism similar. Indeed, mature rats had over 1.5-fold more fat mass, but whole body insulin resistance, as estimated by fasting plasma insulin, was similar to young rats. Mature rats had twice as much AT-TNF activity as the young in both the epididymal (EPI) and retroperitoneal (Retro) fat pads (p < .0005). AT TNF correlated with fasting plasma insulin in Retro only (r = .48, p = .04). AT TNF activity strongly correlated with cell size in both EPI and Retro (r = .79 and .81, respectively, p < .0001). Because cytokines can be regulated at several levels, AT-TNF activity, protein, and mRNA were measured. AT-TNF protein levels were higher in young rats, suggesting that these animals may secrete an inhibitor that reduces AT-TNF activity. There were no significant differences in AT-TNF mRNA between groups. Since TNF has been shown to affect several key genes in tissue culture, mRNA for lipoprotein lipase, hormone-sensitive lipase, and Glut4 were measured. No differences were found between groups. In summary, AT-TNF activity increased in mature animals in relation to adipose cell size. PMID- 9224424 TI - Alterations in atrial natriuretic peptide (ANP) secretion and renal effects in aging. AB - Aging is associated with hypertension and electrolyte disturbances. The purpose of this study was to determine the effect of aging upon secretion and renal actions of atrial natriuretic peptide (ANP). Rats were anesthetized and received tracheal, jugular vein, carotid artery, and bilateral uretheral catheterization. One set of young (2-3 mo) rats (Group 2, n = 9) and one set of old (18-21 mo) rats (Group 4, n = 7) received bilateral atrial appendectomies. Control young (Group 1, n = 8) and old (Group 3, n = 8) rats received a sham appendectomy. All rats were infused (iv) with 6% albumin in Krebs buffer, sufficient to increase blood volume by 15%. Finally, each rat was injected with ANP (1 microgram/kg). Sodium excretion rate (U(Na+)V) in response to volume expansion was significantly decreased in all groups compared to Group 1 (young control, p < .05). All groups demonstrated a striking increase in U(Na+)V with the ANP injection, but the response was greatest in young control rats when factored by body weight (p < .05). There were no significant differences in MAP between the groups, suggesting that the differences in U(Na+)V observed were not the result of hemodynamic factors. Isolated perfused atria from young (n = 9) and old (n = 8) rats were subjected to stretch and endothelin stimulation (50 nM). Atria from young rats showed a dramatic increase in ANP secretion in response to atrial stretch and a further marked increase in secretion in response to endothelin, whereas both of these responses were markedly attenuated in old rats (p < .05). These results suggested that the secretion and renal effects of ANP are impaired in aging. Changes in secretion and actions of ANP in aging could contribute to the development of hypertension or heart failure. PMID- 9224425 TI - Overload-induced C-Myc oncoprotein is reduced in aged skeletal muscle. AB - The C-Myc oncoprotein was examined in anterior latissimus dorsi (ALD) muscles of young (6 weeks) and aged (90 weeks) quail after 0.5 hours to 14 days of stretch. Western analyses of nuclear extracts showed an increase in the C-Myc oncoprotein after 1 h of stretch in young adult birds, and C-Myc remained elevated for 3 days of stretch. The onset and total accumulation of the C-Myc oncoprotein was less in muscles from aged quail as compared to muscles from young adult birds. Immunocytochemical analyses showed that C-Myc was localized in nuclei and averaged 0.2 +/- 0.04 nuclei/muscles fiber cross-section (n/f) in control muscles. C-Myc immunopositive nuclei were more numerous in muscles from young adult birds (1.7 +/- 0.2 n/f) compared to aged birds (1.1 +/- 0.1 n/f) after 2-12 h of stretch. C-Myc positive nuclei declined to 0.7 +/- 0.1 n/f after 3 days of stretch, in muscles from young adult birds; however, this was greater than in muscles from aged birds at the same time point (0.3 +/- 0.04 n/f). Many nuclei that were associated with muscle fibers expressed the C-Myc oncoprotein but did not incorporate bromodeoxyuridine, a marker of DNA synthesis and activated satellite cells. These data show a decreased ability of skeletal muscles from aged quails to initiate a program inclusive of early C-Myc oncoprotein accumulation in response to stretch. PMID- 9224426 TI - Modulation of kinase activities in dauers and in long-lived mutants of Caenorhabditis elegans. AB - Mutant alleles of the genes age-1 and daf-2 that lengthen life span (Age phenotype) of Caenorhabditis elegans cause higher protein kinase (PKA, PKC, PTK) activity levels in senescing worms relative to wild-type. Elevated levels of PKA and PTK were also present in dauer larvae, developmentally arrested juveniles specialized for long-term survival, relative to L3 larvae, the alternative developmental stage. PKC activity was downregulated in dauers of a non-Age control strain and in age-1 mutant dauers, compared to L3 larvae, but similar activities were measured in dauers and L3 larvae of a daf-2 mutant strain. Thus, age-1 and daf-2 mutant worms may express distinct elements of a dauer-specific survival program during adult life. PMID- 9224427 TI - Membrane modifications of red blood cells in Alzheimer's disease. AB - Red blood cells (RBC) from 24 Alzheimer's disease (AD) patients, 18 age- and sex matched nondemented (ND) patients, hospitalized in the same facility for orthopedic problems, and 18 healthy volunteers aged 30-52 years were studied in order to gain insight into the nature of RBC membrane modifications in AD. Significant differences were found between RBC from AD and ND patients or young controls respectively for annexin V-binding (45.5 +/- 18.0% vs 27.1 +/- 14.7 and 2.7 +/- 1.9, p = .003), fraction of glycerol resistant cells (30.8 +/- 11.1% vs 19.6 +/- 6.4 and 10.2 +/- 3.1, p = .026), cell electrophoretic mobility in polymer (1.028 +/- 0.022 microns sec-1 V-1 cm vs 1.046 +/- 0.022 and 1.053 +/- 0.021, p = .02) and only limited significance for the filterability (1.46 +/- 0.12 msec vs 1.58 +/- 0.11 and 1.54 +/- 0.11, p = 0.1). A logistic analysis, using simultaneously several features as independent variables, suggested the combined use of annexinV- binding, glycerol resistance, and cell filterability which allowed the assignment of 95% of patients from this cohort to the right group. A prospective analysis of a larger cohort is required for the estimation of the diagnostic value of this test battery. In addition, the high level of annexin binding is characteristic of a disruption of the phospholipid asymmetry in aged or damaged cells, while the high glycerol resistance combined with low electrophoretic mobility an rigidity characterize young RBC, thus indicating an enhanced turnover of RBC in Alzheimer's disease. PMID- 9224428 TI - Balance and skeletal alignment in a group of elderly female fallers and nonfallers. AB - The purpose of this study was to determine whether sagittal plane posture differed between fallers and nonfallers and to explore the relationship between skeletal alignment and balance in elderly females. Forty-eight women > 65 years of age were recruited from various medical and senior citizen centers. Thirteen of these women were classified as fallers. Spinal posture was measured in standing using an inclinometer, and lower extemity joint angles were assessed in standing using a universal goniometer. The Berg Balance Scale, the Functional Reach Test, end a modified Timed Get Up and Go Test were used to measure balance. Spinal alignment did not differ significantly between fallers and nonfallers; however, knee joint angle was significantly greater in fallers compared to nonfallers. Significant, but low, correlations were found between the inclination of the upper thoracic spine and all three balance measures. Lower thoracic slope and knee joint angle in standing were also weakly related to two of the three balance measures. This study supports the hypothesis that a significant but weak relationship exists between balance and skeletal alignment in elderly females. PMID- 9224429 TI - Sleep disorders and aging: understanding the causes. PMID- 9224430 TI - Correlates of performance-based measures of muscle function in the elderly: the Cardiovascular Health Study. AB - BACKGROUND: It is unknown how much age-related changes in muscle performance represent normal aging versus the effects of chronic disease and life style. We examined the correlates of four performance measures-gait speed, timed chair stands (TCS), grip strength, and maximal inspiratory pressure (MIP)-using baseline data from the Cardiovascular Health Study (CHS), a population-based study of risk factors for heart disease and stroke in persons > or = age 65. METHODS: We analyzed data from the 5,201 CHS participants. Variables were arranged into nine categories: Personal Characteristics, Anthropometry, Physical Condition, Reported Functional Status, Subjective Health, Psychological Factors, Symptoms, Cognitive Status, Habits and Lifestyle, and Prevalent Disease. Independent correlates were identified using stepwise linear regression. RESULTS: The regression models explained 17.7-25.4% of the observed variability. Although age significantly correlated with each measure, it explained little of the variability (< or = 5.7%). Anthropometric features plus physical condition explained 14.0-17.4% of the variability for grip strength and MIP, but 2.8-12.9% of the variability for gait speed and the log of TCS. Subjective health and psychological factors explained 1.8-9.4% of the variability in gait speed and the log of TCS, but < or = 1.2% of the variability in grip strength and MIP. Variables for prevalent disease explained < or = 1.3% of the variability in each measure. CONCLUSIONS: After age 64, age explained little of the variability in muscle performance in a large sample of mostly functionally intact, community dwelling older persons. Complex measures such as gait speed were more associated with subjective factors than were direct measures of strength. Prevalent disease contributed surprisingly little to muscle performance. PMID- 9224431 TI - The association of plasma IL-6 levels with functional disability in community dwelling elderly. AB - BACKGROUND: IL-6 is a multifunctional cytokine that has been shown to increase with age. METHODS: Plasma IL-6 was measured by ELISA in 1,727 community-dwelling elderly subjects whose blood was drawn during the third in-person survey of the Duke Established Populations for Epidemiologic Studies of the Elderly (EPESE). Demographics, functional status (disability), and disease states were determined. Correlations of these factors with IL-6 were analyzed with Spearman's Rho while differences between groups were assessed by Wilcoxon test. RESULTS: IL-6 levels were higher with age (p = .0001) even in this older population (> 70 years). There was a positive correlation between IL-6 and functional disability for each of the functional status measures (p = .0001), as well as a correlation between self-rated health and IL-6. Significantly higher median levels of IL-6 were found in subjects reporting prevalent cancer, heart attack, and high blood pressure, but not diabetes or arthritis. The association between age and functional status with high IL-6 remained when all other variables were controlled, in multivariable analysis. CONCLUSIONS: This association between increased plasma IL 6 levels and functional status suggests that dysregulation of IL-6 may be related to the functional disability seen with aging, and that IL-6 may be useful as a component of an overall marker of health. PMID- 9224432 TI - Assessing functional status: correlation between performance on tasks conducted in a clinic setting and performance on the same task conducted at home. The Salisbury Eye Evaluation Project Team. AB - BACKGROUND: Many studies of functional status in elderly people use performance based measures. There is an underlying assumption that these measures reflect, at least for some tasks, functional abilities in everyday tasks carried out at home. The purpose of this study was to determine the correlation between tasks carried out in the clinic and at home, and the role of visual impairment in performance at either setting. METHODS: We compared the performance of 97 participants in the Salisbury Eye Evaluation (SEE) project at the clinic and at home on eight different tasks: semitandem stand, functional reach, stair climb and descend, inserting a plug, looking up and dialing a telephone number, and reading. RESULTS: The correlations were good for all tasks, with coefficients ranging from .52 to .86. Those with visual impairment were slightly more likely to perform better at home compared to the clinic, although the differences were statistically significant only for the reading task. The most important predictor of performance on any task in the home was performance on the task at the clinic, even after adjusting for age, race, sex, education, and visual impairment. Educational level and visual impairment were consistent predictors of performance in the home for most tests. CONCLUSIONS: We conclude that performances on standardized tasks in the clinic setting do correlate with similar tasks performed in the home, although the relationship is complicated in the presence of visual impairment. PMID- 9224433 TI - The effect of strength and endurance training on gait, balance, fall risk, and health services use in community-living older adults. AB - BACKGROUND: The study tested the effect of strength and endurance training on gait, balance, physical health status, fall risk, and health services use in older adults. METHODS: The study was a single-blinded, randomized controlled trial with intention-to-treat analysis. Adults (n = 105) age 68-85 with at least mild deficits in strength and balance were selected from a random sample of enrollees in a health maintenance organization. The intervention was supervised exercise (1-h sessions, three per week, for 24-26 weeks), followed by self supervised exercise. Exercise groups included strength training using weight machines (n = 25), endurance training using bicycles (n = 25), and strength and endurance training (n = 25). Study outcomes included gait tests, balance tests, physical health status measures, self-reported falls (up to 25 months of follow up), and inpatient and outpatient use and costs. RESULTS: There were no effects of exercise on gait, balance, or physical health status. Exercise had a protective effect on risk of falling (relative hazard = .53, 95% CI = .30-.91). Between 7 and 18 months after randomization, control subjects had more outpatient clinic visits (p < .06) and were more likely to sustain hospital costs over $5000 (p < .05). CONCLUSIONS: Exercise may have beneficial effects on fall rates and health care use in some subgroups of older adults. In community-living adults with mainly mild impairments in gait, balance, and physical health status, short term exercise may not have a restorative effect on these impairments. PMID- 9224434 TI - Leg extension power and walking speed in very old people living independently. AB - BACKGROUND: Leg extension power can be determined as the product of the force and velocity of movement. Its association with maximal walking speed was studied in 131 80- and 85-year-old men and women. METHODS: Leg extension power was measured with the help of a sledge ergometer in a sitting position using a facilitated "jump test." The participant was attached by belts to a sliding chair on rails inclined at 12.6 degrees to the floor. The feet were placed on the force plate attached perpendicularly to the rails, and the knee angle was 90 degrees at the starting position. The participant was advised to extend his or her legs powerfully. The highest value of five to eight attempts was accepted as the result. The results were adjusted for body mass and expressed as watts.kilogram 1. Maximal walking speed was measured in the laboratory corridor over a distance of 10 m. RESULTS: Men and 80-year-old subjects exhibited greater leg extension power and were faster walkers than women and 85-year-old persons. Leg extension power correlated positively with maximal walking speed in all groups: the correlation coefficients were .412 in the 80-year-old men (n = 41, p = .007), .619 in the 80-year-old women (n = 56, p < .001), .939 in the 85-year-old men (n = 8, p = .001), and.685 in the 85-year-old women (n = 23, p < .001). The regression lines for leg extension power and walking speed were coincident, indicating that the power requirements to attain a given walking speed were similar for both sexes. The minimum power threshold for those with a maximal walking speed of 1.30-1.49 m.s-1 was on the order of 4 W.kg-1; a maximal walking speed of 1.50-1.99 m.s-1 required 7 W.kg-1; and for a speed over 2.00 m.s-1 the power threshold was 9.5 W.kg-1. CONCLUSIONS: Their lower average leg extension power may be one of the factors explaining the greater prevalence of mobility problems among women than men. PMID- 9224435 TI - The effects of two types of cognitive tasks on postural stability in older adults with and without a history of falls. AB - BACKGROUND: This study used a dual task design to investigate the effects of two different types of cognitive tasks on stability (as measured by center of pressure displacement) in young vs older adults with and without a history of falls. METHODS: Two secondary cognitive tasks, a sentence completion and a visual perceptual matching task, were used to produce changes in attention during quiet stance under flat vs compliant surface conditions in 20 healthy young adults, 20 healthy older adults, and 20 older adults with a history of imbalance and falls. Postural stability was quantified using forceplate measures of center of pressure (COP). Speed and accuracy of verbal response on the cognitive tasks were also quantified. RESULTS: During the simultaneous performance of a cognitive and postural task, decrements in performance were found in the postural stability measures rather than the cognitive measures for all three groups. While no differences were found between the young adults and the older healthy adults on the firm surface, no task condition, when task complexity was increased (either through the introduction of a secondary cognitive task, or a more challenging postural condition such as standing on the compliant surface), significant differences in postural stability between the two groups became apparent. In contrast to the young and healthy older adults, postural stability in older adults with a history of falls was significantly affected by both cognitive tasks. CONCLUSION: Results suggest that when postural stability is impaired, even relatively simple cognitive tasks can further impact balance. Results further suggest that the allocation of attention during the performance of concurrent tasks is complex; depending on many factors including the nature of both the cognitive and postural task, the goal of the subject and the instructions. PMID- 9224437 TI - Causes of death in geriatric patients: a cross-cultural study. AB - BACKGROUND: The elderly are living longer and causes of death are shifting. At the same time, autopsy rate is at, or near, its lowest in history, compounded by an even lower interest in geriatric autopsies. Thus, the prevalent cause of death in this age group remains poorly studied. METHODS: In a retrospective study, the autopsy protocols of 440 70-year-old or older patients from the Houston Veterans Affairs Hospital and 321 80-year-old or older patients from the II*Institute of Pathology in Prague (Czech Republic) were reviewed in order to establish a correct cause of death. The autopsy diagnosis was correlated with the prosectors' description of pathological findings in the protocol. In questionable cases or discrepancies, the patient's clinical chart and/or the histological autopsy slides were also reviewed. RESULTS: The distribution of death by infections and cardiac disorders each accounted for one-third of all deaths. Congestive heart failure prevailed in the over 80-year-olds, and myocardial infarcts prevailed in the younger patients. The number of deaths due to malignancy dropped from 25% in those 70-79 years old to about 10% in the elder patients. Central nervous system disorders were frequent as an underlying disease, but were not common as a cause of death. The findings were similar in both series, thus supporting their accuracy. CONCLUSION: Our findings bring into question the accuracy of reported causes of death in the elderly. With increasing age, differences appear in the levels of mortality and morbidity for various disease categories. This study underlines the need for more baseline data for older people which can be obtained only by more and well-performed autopsies. PMID- 9224436 TI - Characteristics of depressive symptoms in elderly urban and rural African Americans. AB - BACKGROUND: Despite considerable progress in the epidemiology of late life depression, little data have been documented in the scientific literature on depressive symptoms among elderly African Americans. The present investigation identifies characteristic symptoms of depression in African American community resident elders. METHOD: Ninety-six African American men and women aged 60 years and older, with equal representation from urban and rural counties in west Tennessee, composed the sample. The sample was stratified in each of the two counties into three age categories; 60-69, 70-79, and 80 and older. Data from the Center for Epidemiological Studies-Depression scale were compared with the association of medical illness, medication use, social network, level of physical function in activities of daily living, and demographic characteristics. RESULTS: Residents screening positive for the presence of depressive symptoms showed an increased report of hypertension (p < .036), arteriosclerosis (p < .035), and circulatory problems (p < .008). There was an increased report of symptoms of depression among those who had six or more different chronic illnesses (p < .001) and among those who reported using four or more different prescription medications in the past month for chronic illnesses (p < .015). Regression analyses of data indicated that medical illness (p < .001) and social network (p < .041) were the most important predictors of depressive symptoms among residents. CONCLUSIONS: Considering the projected increase of African Americans reaching age 60, and because depressive illness is an important public heath concern, early identification of salient risk factors for depression is critical in instituting early intervention programs for the ethnic minority elderly population. PMID- 9224438 TI - Altered neuroendocrine control of GH secretion in normal women of advanced reproductive age. AB - BACKGROUND: Previous studies have suggested that the neuroendocrine control of growth hormone (GH) secretion changes with increasing age in women with normal menstrual cycles and sex steroid levels. METHODS: In order to verify this hypothesis, 8 younger (22-32 years) and 8 older (41-45 years) women with normal menstrual function and gonadal steroid levels were tested with the serotonergic agent sumatriptan (6 mg in a subcutaneous bolus), the GABAergic agonist sodium valproate (800 mg orally), the dopaminergic compound L-Dopa (500 mg orally) and placebos. Furthermore, all women were tested with GH-releasing hormone (GH-RH 1 microgram/kg body weight in an intravenous (i.v.) bolus) to determine whether GH secretion in response to its specific releasing factor was preserved. Serum GH levels were recorded over 2 hours in all tests and IGF-I levels in basal samples. RESULTS: Plasma IGF-I concentrations and the GH responses to sumatriptan, sodium valproate and L-Dopa were significantly lower in older than in younger women. Also, the GH-RH-induced GH response was significantly lower in older than in younger subjects. When peak GH responses to releasing stimuli were compared with age, significant negative correlations were found in all tests. CONCLUSIONS: These data did not show a specific neurotransmitter change underlying defective GH secretion in older aged reproductive women. On the other hand, the results indicated that age-related changes in the secretory machinery of GH, such as a reduced pituitary sensitivity to GH-RH and/or a reduction in the pituitary GH secretory capacity, affect women during the last years of the reproductive period. PMID- 9224439 TI - Behavioral treatment of depression in dementia patients: a controlled clinical trial. AB - The current study is a controlled clinical investigation of two nonpharmacological treatments of depression in patients with Alzheimer's disease. Two active behavioral treatments, one emphasizing patient pleasant events and one emphasizing caregiver problem solving, were compared to an equal-duration typical care condition and a wait list control. Seventy-two patient-caregiver dyads were randomly assigned to one of four conditions and assessed pre-, post-, and at 6 months follow-up. Patients in both behavioral treatment conditions showed significant improvement in depression symptoms and diagnosis as compared with the two other conditions. These gains were maintained at 6-month follow-up. Caregivers in each behavioral condition also showed significant improvement in their own depressive symptoms, while caregivers in the two other conditions did not. Results indicate that behavioral interventions for depression are important and effective strategies for treating demented patients and their caregivers. PMID- 9224440 TI - The measurement of mood states in older adults. AB - This study examined the reliability and validity of the Profile of Mood States (POMS) questionnaire when administered to 479 community-dwelling older adults. Factor analysis replicated the original factor structure identified in young adult samples, suggesting that older adults adopt the same underlying constructs of mood when responding to the POMS. There was strong support for concurrent validity, and this instrument was able to discriminate between healthy adults and patients with known mood disturbance. Excellent internal consistency of POMS subscales and very good retest reliability were noted. A significant age-related decline in self-reported mood states emphasizes the need for age-specific norms, although response bias rather than age itself was shown to account for this decline. Overall, the POMS can be recommended as a reliable and valid measure of mood states in older adults, although it is clear that normative values for this test vary considerably over the age spectrum. PMID- 9224441 TI - Asking the age question in elderly populations: a reverse record check study. AB - In two large-scale surveys among elderly respondents we evaluated the accuracy of answers obtained to three differently formulated age questions. Respondents included 6,149 individuals aged 65-86 living in The Netherlands. Because criterion age data were available from different sources, it was possible to compare the respondent's reported age with his or her actual age. Refusal rates were low for all three questions. Both age and cognitive capabilities influenced accuracy of the answers to the age questions. The results indicated that the most accurate data were obtained with the question, "What is your date of birth?" in combination with interview date. PMID- 9224442 TI - Older and younger adults use a functionally identical algorithm to select items for restudy during multitrial learning. AB - We investigated whether aging affects several components of how people select items for study during multitrial learning. Younger and older adults studied paired-associate items and then made delayed judgements of learning (JOLs). Immediately after making a JOL for an item, some participants decided whether to restudy the item on subsequent trials; for other participants, the computer selected for restudy the items that had been judged as least-well learned. Next, paired-associate recall occurred, which was followed by restudy-test trials. As expected, age differences occurred in recall on the first trial, and this difference was propagated across trials. In contrast to the hypothesis that older adults would be more conservative in selecting items, both age groups selected to restudy (a) the items that they had rated as least-well learned and (b) the majority of items that would not be recalled on the first trial. Comparisons between participants who self-selected items vs the groups in which the computer controlled selection also converged on the conclusion of age equivalence in processes underlying item selection. PMID- 9224443 TI - Adult age-group differences in recall for the literal and interpretive meanings of narrative text. AB - This study examined age differences in recall for the literal and interpretive meanings of narrative text. Following presentation of one of two stories rich in both literal and interpretive content, younger (mean age = 19.2 years) and older (mean age = 72.2 years) adults were asked to retell and to interpret the story. Response task order was counterbalanced across participants. When asked to retell a story as close to the original as possible, the younger adults recalled more of the literal propositional content than did the older adults in the retell-first, although not in the interpret-first, condition. In addition, both older and younger adults recalled more of the main ideas (gist) relative to the details. When asked to interpret the same story, more older than younger adults produced deep and synthetic representations of the story's interpretive meanings. In addition, there was a clear preference among the older age group for deep synthetic responding. Although more younger than older adults produced analytic interpretations, within the younger group there was no clear preference for either an analytic or a deep-synthetic style. PMID- 9224444 TI - Positive and negative social exchanges: weighing their effects in later life. PMID- 9224445 TI - Women's caregiving and paid work: causal relationships in late midlife. AB - Care of an ill or disabled family member or friend is disproportionately done by women and typically is done in late midlife. Because this is-also a time in the life course when women's labor force participation peaks, many women faced with caregiving demands have to decide how to balance them with their employment. In this study we use the National Longitudinal Survey (NLS) of Mature Women to examine the causal relationship between employment and caring for an ill or disabled friend or relative over a three-year period. We find that employment does not affect whether or not women start caregiving, but that women who do start are more likely to reduce employment hours or stop work. Thus, the causal relationship between employment and caregiving in late midlife is largely unidirectional, with women reducing hours to meet caregiving demands. PMID- 9224446 TI - The influence of caregiving and employment on the voluntary activities of midlife and older women. AB - One factor thought to contribute to higher levels of stress among caregivers is the restriction on personal time and leisure activities that they feel. We use data from the National Survey of Families and Households (NSFH) to examine the influences that caregiving, the relationship of the care-recipient with the caregiver, and the intensity of the caregiving have on women's participation in personal, family-centered, and community activities. We find that care-giving regardless of age, does not reduce the frequency of participation in voluntary activities. In fact, among younger women, some types of caregiving are associated with significantly higher levels of participation. Based on our results, we conclude that caregiving does not necessarily result in a "loss of self." Caregivers may be using outside activities as a way to relieve the stress of the caregiving tasks; caregivers may be particularly adept at balancing roles, or most caregiving may not be at a level of intensity sufficient to interfere with other activities. PMID- 9224447 TI - The effects of positive and negative social exchanges on aging adults. AB - This study tested various models of the effects of positive and negative exchanges on positive and negative affect using structural equation modeling. Based on a probability sample of middle-aged and older adults, the relationships between social exchanges and psychological well-being were examined both within the total sample and within subgroups of individuals who had experienced few vs many life events. Within the general population, the Domain Specific Model resulted in the best fit. That is, positive exchanges were associated with positive affect, and negative exchanges were associated with negative affect. However, among the subgroup that had experienced more life events, there was a significantly stronger relationship between negative exchanges and negative affect. These findings suggest that, to understand the effects of social exchanges, it is important to consider the context of life events. PMID- 9224448 TI - Social support and depressive symptoms: differential patterns in wife and daughter caregivers. AB - This cross-sectional study examined how three types of social support-social participation, emotional support, and caregiving support-were related to depressive symptoms in wives caring for their elderly husband and daughters caring for their elderly parent. We investigated whether different dimensions of social support affect mental health via different mechanisms and whether the context in which the support is needed and received will temper its effects. We found that social participation had a main effect on depressive symptoms for daughters but not for wives. Emotional support buffered the stress emanating from the husband's behavior problems for wives. For daughters, emotional support buffered the stress emanating from both the behavior problems and the ADL/IADL limitations of the parent care recipient. Using caregiving as the example, our data suggested that social support does not have uniform effects; rather, the type of stressor, the type of social support, and the individual context interact to result in the specific effect of support. PMID- 9224449 TI - Age and stage of readiness for smoking cessation. AB - Using the 1992 National Health Interview Survey Cancer Control Supplement, relationships were analyzed between stage of readiness for smoking cessation and background characteristics, smoking behaviors, and smoking-related attitudes among smokers aged 18-29, 30-49, and > or = 50 years. For each age group, an ordinal logistic regression model was computed to identify correlates of readiness to quit. The youngest smokers had attitudes most favorable to being ready to try to quit smoking. For smokers aged 30-49, the influence of a medical provider and perceived health effects of smoking were important correlates of readiness. Among smokers 50 and older, those with realistic health consequences of smoking and those who perceived smoking as addictive were more likely to be ready to quit. The effectiveness of smoking cessation programs might be improved by matching interventions to a smoker's age and stage in the smoking cessation process. PMID- 9224450 TI - Transitions in chronic low back pain in Japanese older adults: a sociomedical perspective. AB - This study examines the patterns and determinants of chronic low back pain over a three-year period among older adults in Japan. We tested our model based on a sociomedical perspective, using a two-wave national probability sample survey of persons aged 60 and older (N = 2,200) conducted in 1987 and 1990 in Japan. At baseline, the prevalence of chronic low back pain was 18 percent. Among those who were free of back pain at baseline, the probabilities of onset, death, and nonresponse were 13 percent, 7 percent, and 10 percent. Among those who had back pain at baseline, the probabilities of recovery, death, and nonresponse were 43 percent, 8 percent, and 9 percent, respectively. Our multinomial logistic regression analysis supports our thesis that societal factors (age, gender, education, and social relationships) affect transitions in chronic back pain not only directly, but also indirectly through mediating health and health behavior factors. The results suggest that social relationships have both favorable and unfavorable effects on chronic low back pain. PMID- 9224451 TI - Abscessed hydroxyapatite orbital implants. PMID- 9224452 TI - Office repair of retinal detachment. PMID- 9224453 TI - Aspergillus endophthalmitis in orthotopic liver transplant. PMID- 9224454 TI - Patient self-management skills influence the course of glaucoma. PMID- 9224455 TI - The intrastromal corneal ring segments. Phase II results for the correction of myopia. AB - OBJECTIVE: The purpose of the study was to evaluate the safety and efficacy of the intrastromal corneal ring segments (ICRS) for the correction of myopia. DESIGN: A 2-year phase II clinical trial of ICRS was initiated in May 1995. The investigational plan specifies that 150 patients with sighted eyes, requiring myopic corrections from -1.00 to -6.00 diopters (D), will each receive ICRS in 1 eye. The patient population will be divided into approximately five patients per ICRS thickness (0.25, 0.30, 0.35, 0.40, and 0.45 mm) per site. Six investigational sites are participating in the trial. PARTICIPANTS: Fifty-nine men and 43 women requiring myopic corrections were enrolled at four U.S. investigational sites. These 102 patients each received the ICRS product in 1 eye. INTERVENTION: Correction of myopia. MAIN OUTCOME MEASURES: Efficacy of ICRS was assessed with respect to the trial endpoints of predictability of refractive effect, uncorrected visual acuity (UCVA), stability of UCVA, maintenance of best spectacle-corrected visual acuity and stability of refractive effect. RESULTS: As shown by the available month-3 data (99 patients; all device thicknesses), 95 (96%) of 99 patients had a UCVA of 20/40 or better. Ninety-eight (99%) of 99 patients were within 2 lines of their preoperative best spectacle-corrected visual acuity. The average change (with standard error) in cycloplegic refraction (spherical equivalent) achieved by ICRS thickness was -1.27 +/- 0.09 D (0.25 mm), -2.13 +/- 0.16 D (0.30 mm), -2.56 +/- 0.15 D (0.35 mm), -3.77 +/- 0.37 D (0.40 mm) and -4.16 +/- 0.24 D (0.45 mm). Seventy-seven percent (76/99) of the patients were within +/-1.00 D of their intended correction. When the ICRS was removed in two cases, both patients returned to within 0.75 D of their preoperative manifest refraction. CONCLUSIONS: The ICRS appears to be a viable and effective alternative for the treatment of myopia. Additionally, as indicated by the explant data, the ICRS's refractive effect may be reversible upon removal of the device. PMID- 9224456 TI - Ocular integrity after refractive procedures. AB - PURPOSE: The purpose of the study was to determine the integrity of human eyes after refractive procedures. METHODS: Whole human globes underwent either radial keratotomy (RK) with eight incisions, automated lamellar keratoplasty (ALK), photorefractive keratectomy (PRK), or excimer laser assisted in situ keratomileusis (LASIK). Eyes then were subjected to quantitatively increasing levels of trauma until rupture occurred. RESULTS: All eyes operated on required less energy to rupture as compared with that of control eyes. The mean number of trials required for rupture is as follows (energy doubled with each successive trial): normal, 4.29; LASIK, 3.80; ALK, 3.67; PRK, 3.60; and RK, 2.83. The level of energy required to rupture normal, ALK, PRK, and LASIK eyes was not significantly different. All RK eyes ruptured at incisions. Most ALK, PRK, and LASIK eyes ruptured near the flap edge or limbus. Most normal eyes ruptured with both corneal and scleral involvement. Age of tissue donors at the time of death and time elapsed between death and procedure were not significantly different between groups (P = 0.88 and 0.79, respectively). CONCLUSIONS: The energy required to rupture ALK, PRK, and LASIK eyes is not significantly different from that for normal eyes. The RK eyes ruptured with significantly less energy than did normal eyes. All RK eyes ruptured at incision sites. PMID- 9224457 TI - Postoperative cellular reaction on various intraocular lens materials. AB - PURPOSE: The presence of cellular deposits on the surface of intraocular lenses (IOLs) is a manifestation of: (1) the breakdown of the blood-aqueous barrier produced by surgery; and (2) foreign body reaction induced by lens implantation. The purpose of this study was to assess the presence of cellular deposits on the surfaces of various IOL materials. METHODS: Fifty patients scheduled for cataract surgery were randomized into five groups of ten patients each and received IOLs of the following materials: conventional polymethylmethacrylate (PMMA), surface passivated PMMA, heparin-surface modified PMMA, poly-hydroxyethylmethacrylate (HEMA) hydrogel and silicone. Patients were examined at 7 days, 30 days, 90 days, and 180 days after surgery. All eyes were observed first via slit-lamp and then using a contact specular microscope for photographic documentation. RESULTS: Small, spindle-shaped cells were observed on all IOLs in the early postoperative period. Epithelioid cells appeared approximately 30 days after surgery on all PMMA IOLs, but most particularly on conventional PMMA IOLs. No cells were observed on poly-HEMA and silicone IOLs. CONCLUSIONS: The decreased number of epithelioid cells discovered in the early postoperative period may indicate a reduction in the inflammatory process induced by surgery. The permanence of epithelioid cells on IOL surfaces may be a sign of foreign body reaction. The results of this study indicated that poly-HEMA and silicone IOLs showed fewer cellular deposits than PMMA IOLs, suggesting that they may be better tolerated than PMMA IOLs. PMID- 9224458 TI - Cataract after vitrectomy in young patients. AB - OBJECTIVE: The purpose of the study was to evaluate the occurrence of cataract in young patients after pars plana vitrectomy. DESIGN: The authors reviewed the medical records for previtrectomy and postvitrectomy lens changes in patients younger than 30 years of age at the Kellogg Eye Center, University of Michigan. PARTICIPANTS: Forty-nine patients (50 eyes) younger than 30 years of age (mean age, 23.5 years; range, 5 months-30 years) underwent phakic vitrectomy over a 12 year period. MAIN OUTCOME MEASURES: Cataracts were categorized as posterior subcapsular, nuclear sclerotic, or cortical. Cataracts also were graded as mild (1+), moderate (2+), or severe (3+). RESULTS: In this series of young patients, vitrectomy was performed for a wide range of ocular conditions, including trauma and complicated retinal detachment. Postvitrectomy cataract developed in 29 patients (60%). Eighteen patients (36%) had visually significant cataract on long term follow-up (mean follow-up, 29.7 months; range, 6 months-13 years). The most common cataract was posterior subcapsular (57%), followed by nuclear sclerosis (23%), a combination of both (17%), and cortical cataract (3%). Patients with gas filled eyes had a significantly higher rate of cataract formation than patients with fluid-filled eyes (P < 0.05). CONCLUSIONS: Postvitrectomy cataracts were more common in the authors' series compared with those of previous reports on young patients. Cataracts were most often posterior subcapsular and were significantly associated with the use of intraocular gas. The occurrence of postvitrectomy cataract appears to be higher in patients undergoing vitrectomy for complex ocular conditions, regardless of age. PMID- 9224459 TI - Pediatric cataract management with variations in surgical technique and aphakic optical correction. AB - PURPOSE: The purpose of the study was to compare the results of three techniques of cataract surgery in children. Two methods included intraocular lens (IOL) implantation and one used contact lens correction of aphakia. DESIGN: Nonrandomized clinical trial. PARTICIPANTS: Seventy-seven eyes of 50 children between the ages of 2 1/2 and 16 years who had cataract surgery for the treatment of uncomplicated cataract. INTERVENTION: Thirty-one eyes underwent a "conventional" style of implantation, and a "phaco-style" of surgery was used in 24 eyes. A contact lens was used as the primary means of aphakic correction in 22 eyes. MAIN OUTCOME MEASURES: The visual results and complications of each type of surgery were compared. RESULTS: Corrected visual acuities did not differ significantly between the three groups 6 months after surgery. The incidence and type of complications were significantly different. Better lens centration, less long-term iris changes, or wound-related problems were observed with "phaco style" modification of the technique of IOL insertion. CONCLUSIONS: Pediatric IOL insertion eliminated the need for contact lens wear and did not lead to a significantly different corrected visual acuity 6 months after surgery compared with lensectomy with contact lens correction. Adoption of some of the techniques of modern small-incision cataract surgery for pediatric IOL procedures produces a significant reduction in postoperative anterior segment complications compared with a standard limbal approach. Such modifications allow pediatric IOL insertion to be a safe alternative for the correction of pediatric aphakia. PMID- 9224460 TI - Resolution of astigmatism after surgical resection of capillary hemangiomas in infants. AB - OBJECTIVE: The goal of the study was to assess the affect of early surgical resection of capillary hemangiomas on the induced astigmatism of infants. DESIGN: Cohort study. PARTICIPANTS: Three infants younger than 9 months of age are included. INTERVENTION: Total resection of the astigmatism-inducing hemangiomas was performed. MAIN OUTCOME MEASURES: Refractive change in the eye operated on was measured and the cosmetic results were observed. RESULTS: Preoperative and postoperative cylinder reduction in the three patients were 8 diopters (D) to 0 D, 3 D to 0 D, and 4.5 D to 1 D, respectively. All patients had excellent cosmetic results, and there were no postoperative complications. CONCLUSIONS: Surgical resection of adnexal hemangiomas in carefully selected infants can lead to excellent cosmetic results. If the mass-induced pressure on the infant sclera is relieved at a young enough age, anisometropia and resultant amblyopia can be eliminated. PMID- 9224461 TI - Post-traumatic corneal mucormycosis caused by Absidia corymbifera. AB - OBJECTIVE: The purpose of the study was to report a case of mycotic keratitis caused by the organism Absidia corymbifera (class Zygomycetes, order Mucorales, family Mucoraceae). DESIGN: Case report. PARTICIPANT: A healthy 37-year-old farmer scratched his left cornea on a galvanized nail while working in his barn. Within 24 hours, an infiltrate in the interior cornea developed that advanced superiorly, reducing the vision to hand motion by the following day. He was treated with topical and systemic antibiotics and antifungal medications, but the infiltrate spread to the adjacent nasal limbus. INTERVENTION: An 11-mm penetrating keratoplasty was performed with an adjacent nasal 7-mm superficial lamellar sclerectomy. MAIN OUTCOME MEASURES: Pathologic examination of the keratoplasty specimen. RESULTS: Corneal cultures grew A. corymbifera. The organisms were identified in tissue sections by light, fluorescent, electron, and immunoelectron microscopy. CONCLUSIONS: The authors believe that this is the first reported case of keratitis caused by an Absidia species and, as such, represents an unusual form of mucormycosis in an otherwise healthy individual. PMID- 9224462 TI - Neovascular glaucoma developing after uncomplicated cataract surgery for heavily irradiated eyes. AB - PURPOSE: The purpose of the study was to evaluate the effect of cataract surgery and postoperative panretinal photocoagulation (PRP) on the development of neovascular glaucoma (NVG) in heavily irradiated eyes. PATIENTS AND METHODS: The authors performed a retrospective study on the incidence of NVG in 90 eyes that had received megavoltage external beam irradiation at a retinal dose of 56 to 80 Gy and that had at least 48 months of follow-up. These eyes were categorized into different groups depending on whether cataract surgery or PRP was performed. Rates and proportions of NVG occurring in these groups were compared and analyzed with one-tailed Fisher's exact test. RESULTS: The incidence of NVG was significantly higher in patients who underwent cataract surgery without postoperative PRP (P < 0.01). Neovascular glaucoma did not develop in any patient who underwent cataract surgery and PRP. CONCLUSIONS: Cataract surgery may accelerate the development of NVG in heavily irradiated eyes. Photoablation of ischemic retina is recommended before cataract surgery or soon thereafter if cataract density precludes laser treatment. PMID- 9224463 TI - Ultrasound biomicroscopy in infantile glaucoma. AB - BACKGROUND: Glaucoma in infants has many causes: Evaluation of the anatomy of the anterior segment of eyes with infantile glaucoma may help to determine the pathogenesis of an infant's disease and influence therapeutic decisions. METHODS: Eleven eyes of six infants with glaucoma were evaluated with ultrasound biomicroscopy (UBM) to evaluate the anatomic characteristics and relationships of the anterior segment structures. RESULTS: The anterior chamber angle, iris, lens, ciliary body, and posterior chamber angle could be imaged in detail. Elongated and anteriorly placed ciliary processes were noted in all eight eyes with trabeculodysgenesis. There were no apparent anomalies in the trabecular meshwork, or anterior chamber. In three eyes with dense corneal opacities, ultrasound biomicroscopy showed severe anterior segment disorganization and thin central corneas with posterior corneal excavation. CONCLUSIONS: Ultrasound biomicroscopy is a useful non-invasive method for evaluating infants with glaucoma in cases with corneal opacities. This information can help in surgical planning for glaucoma management. PMID- 9224464 TI - Long-term follow-up of initially successful trabeculectomy. AB - OBJECTIVE: The purpose was to study the long-term outcomes of primary trabeculectomies that were successful at 1 year. DESIGN: A retrospective study of patients with various types of glaucoma who had trabeculectomies that were successful at 1 year and who had a follow-up of at least 10 years. PARTICIPANTS: There were 40 patients (40 eyes) who had primary trabeculectomies that were successful at 1 year and who had a follow-up range of 10 to 21 years. INTERVENTION: Control of intraocular pressure (IOP) and disease progression was evaluated at 5, 10, and 15 years and at the last obtainable follow-up. MAIN OUTCOME MEASURES: Successful control of IOP was defined as IOP less than 21 mmHg or a reduction of 33% if preoperative IOP was less than 21 mmHg. Successful control of disease progression was defined as stable cup-disc ratios determined by examination, or color photographs or both, as well as stable visual fields. RESULTS: If an eye was considered successful by IOP at 1 year, the probability of successful control of IOP was 82% at 5 years and 67% at 10 and 15 years. If an eye was considered successful by IOP at 1 year, the probability of successful control of disease progression at 5 years was 77%, at 10 years 61%, and at 15 years 48%. If an eye did not require further glaucoma surgery at 1 year, the probability that it still would not need further surgery at 5 years was 90%, at 10 years 75%, and at 15 years 67%. Forty percent of eyes had cataract extraction by the time of last follow-up examination. CONCLUSIONS: Loss of IOP control and progression of glaucomatous damage occurs over time despite initial success at 1 year. PMID- 9224465 TI - Analysis of reliability indices from Humphrey visual field tests in an urban glaucoma population. AB - PURPOSE: Visual field assessment is extremely important in glaucoma management, but interpretation is affected by the quality of the patient's performance. The authors have investigated the reliability of visual field performance by a randomly selected sample of the chronic glaucoma population at an urban tertiary care practice. METHODS: Patient reliability in Humphrey automated visual field testing was studied in 106 randomly selected chronic open-angle glaucoma patient charts, which provided 768 tests (mean, 7.2 +/- 4.8 fields; range, 2-18 fields). Reliability criteria were established as less than 20% fixation losses, less than 33% false-negative error, and less than 33% false-positive error, as recommended by Humphrey Instruments, Inc (San Leandro, CA). RESULTS: Patients performed reliably in 61% of right eye fields, 58% of left eye fields, and 59.5% overall. Of the 106 patients, only 35 (33%) were always reliable in both eyes, whereas 8 (7.5%) were always unreliable in both eyes. The most common cause of unreliability was fixation loss (39%), whereas false-positive error (5%) and false-negative error (9%) were less frequent. A more severely depressed mean deviation correlated significantly with poorer performance on the three reliability indices, with false-negative error having the greatest correlation, followed by fixation loss and false-positive error. Corrected pattern standard deviation correlated closely only with false-negative error. Prolonged test time also correlated with all three reliability indices. Age was a significant factor for fixation loss but not for false-negative or false-positive error. CONCLUSIONS: The authors conclude that fewer than two thirds of the Humphrey visual fields were reliable with the authors' urban tertiary care population of patients with glaucoma. Relaxing the fixation loss criterion to less than 33% improved the rate of reliability to approximately 75%. The severity of glaucomatous visual field defects, test time, and age were identified as factors influencing the reliability of the Humphrey visual fields. PMID- 9224466 TI - Effect of surgery on visual field progression in normal-tension glaucoma. AB - PURPOSE: The effect of intraocular pressure-lowering surgery on the rate of visual field progression in normal-tension glaucoma (NTG) was studied. METHODS: Seventeen patients with NTG who underwent trabeculectomy in one eye for worsening visual field loss were included in the first part of the study. All patients had Humphrey 24-2 visual fields at the rate of 2-3 fields per year. Pointwise linear regression analysis of the visual field data was done separately for the preoperative and postoperative periods. This was performed for both operated and fellow eyes. The mean slope (MS), which indicates rate of visual field progression, was calculated. Change in MS was correlated with change in intraocular pressure (IOP). For the second part of the study, 11 patients who had a minimum of 4 visual fields and 18 months of follow-up before surgery were identified. Using the preoperative fields, the rate of sensitivity loss for each visual field location in the operated eye was ascertained for every patient. This rate of loss was extrapolated to generate the expected visual fields, assuming an unchanged rate of progression. The mean sensitivity of the expected visual field was compared with that of the actual field at the last follow-up. RESULTS: The MS in the operated eyes improved from -2.97 +/- 3.21 (mean +/- SD) in the preoperative period to 0.53 +/- 3.83 (P < 0.005; Student's t test) postoperatively. In the fellow eyes the MS changed from -1.78 +/- 2.55 to -1.43 +/- 3.01 (P = 0.754). There was a weak correlation between change in MS and percentage IOP decrease (correlation coefficient 0.39). The difference in mean sensitivity between the expected and actual visual fields was -3.72 dB (P = 0.002), and was better in the actual field. CONCLUSIONS: In this study, surgical lowering of IOP resulted in a slower rate of visual field loss in the operated eye. PMID- 9224467 TI - Evaluation of coexisting optic nerve head drusen and glaucoma with optical coherence tomography. AB - OBJECTIVE: Optic nerve head drusen often make evaluation of the nerve head difficult to interpret. In addition, visual field defects are known to occur in patients with optic disk drusen, resembling glaucomatous damage. The authors report two cases of coincident optic nerve head drusen and glaucoma, in which the use of optical coherence tomography (OCT) in evaluating the nerve fiber layer was beneficial. PARTICIPANTS: Two patients with both optic nerve head drusen and glaucoma, one with primary open angle glaucoma, the other with pseudoexfoliation glaucoma were evaluated. Both patients had asymmetric optic disk drusen, with clinically visible drusen only in one eye. INTERVENTION: Ophthalmologic examination, color and red-free photography, automated Humphrey visual field testing and OCT were performed. RESULTS: Nerve fiber layer loss as measured by OCT was found to be greater than expected by the appearance of the optic nerve head and red-free photography, with visual fields consistent with findings in case 1. In case 2, visual fields were full, despite nerve fiber layer thinning seen by OCT and red-free photography. CONCLUSIONS: There can be significant nerve fiber layer thinning in patients with both glaucoma and optic disk drusen, despite the appearance of the optic nerve head in these patients. The cup margin may be obscured by the drusen, giving rise to a falsely full-appearing disk. In such cases, OCT may provide a useful means to quantitatively measure the nerve fiber layer thickness and to aid in the management of these patients by detecting nerve fiber layer thinning earlier than would otherwise be possible. PMID- 9224468 TI - Application of rapid scanning retinal thickness analysis in retinal diseases. AB - PURPOSE: The authors have further developed their method of retinal thickness analysis to rapidly generate multiple optical cross sections of the retina and provide thickness maps at the posterior pole. The potential use of this method was evaluated in a number of macular disorders. METHODS: A commercial prototype of the scanning retinal thickness analyzer was used to examine patients with a variety of macular diseases. A laser slit beam was projected on the retina and scanned across a 2- X 2-mm retinal area in 200 to 400 msec. The images of the intersection of the laser slit beam with the retina were recorded digitally and used for visualization of disease. Nine scans were combined, and an operator-free algorithm generated a three-dimensional thickness map at the posterior pole. RESULTS: Cysts could be visualized in macular edema associated with diabetes mellitus and with retinal vein occlusion. The retinal thickness map quantitated the location, extent, and height of the edema. In serous detachment, the extent and the height of the retinal pigment epithelial elevation could be documented. In cases of suspected macular holes and pseudoholes, the diagnosis was considered more reliable than with conventional biomicroscopy. The extent of epiretinal membranes, the sites of adherence, and associated intraretinal cystic changes were identified. In glaucoma, the anatomic course of localized loss of neuronal retinal tissue could be traced. CONCLUSIONS: Scanning retinal thickness analysis provided multiple optical cross sections of the retina and yielded information useful in the diagnosis and monitoring of macular diseases. The three-dimensional thickness map provided quantitative information that may be useful for clinical management. PMID- 9224469 TI - Choroidal metastases from renal cell carcinoma. AB - BACKGROUND: Choroidal metastases from renal cell carcinoma are uncommon. The authors investigated the clinical characteristics of patients with renal cell carcinoma in whom choroidal metastases developed. METHODS: The clinical records of five patients with histopathologically confirmed renal cell carcinoma and choroidal metastases were reviewed retrospectively. RESULTS: In four patients, choroidal metastases were either the sole initial manifestation of disease or were the initial manifestation of metastatic disease. The interval from nephrectomy to the onset of ocular signs ranged from 6 to 18 years. A reddish orange appearance of the tumor was present in two patients, but no pathognomonic features distinguishing these tumors from other choroidal metastases were identified. CONCLUSIONS: Ocular metastases may precede the diagnosis of renal cell carcinoma or may follow it by years or decades. This interval between its ocular and systemic presentation may be so prolonged as to obscure the relation between the choroidal metastases and the primary tumor. In patients with amelanotic or reddish choroidal lesions without known metastatic disease, evaluation of the kidney may be warranted as part of a metastatic workup to exclude metastatic renal cell carcinoma. PMID- 9224470 TI - The management of giant retinal tears using perfluoroperhydrophenanthrene. A multicenter case series. Vitreon Collaborative Study Group. AB - OBJECTIVE: The purpose of the study was to determine the predictors of success and evaluate the use of perfluoroperhydrophenanthrene as an intraoperative and postoperative tool in the management of giant retinal tears in a multicentered collaborative study. DESIGN: Multicentered prospective case series. PARTICIPANTS: Twenty-three centers consecutively enrolled 162 eyes of 161 patients with retinal tears 90 degrees or greater in circumferential extent. INTERVENTION: Perfluoroperhydrophenanthrene was used as an intraoperative surgical adjunct in all cases and left after surgery in 16 eyes (9.9%). MAIN OUTCOME MEASURES: Retinal reattachment and visual acuity. RESULTS: Intraoperative reattachment was achieved in 158 eyes (97.5%); 147 eyes (90.7%) remained attached at their most recent follow-up. Seventy-nine eyes (48.8%) experienced an improvement in their visual acuity, 26 eyes (16.0%) remained unchanged, and 57 (35.2%) worsened. Recurrent retinal detachment occurred in 80 patients (49.4%). Other significant postoperative complications included cataract formation in 20 (39.2%) of 51 eyes, macular pucker in 12 (7.4%), corneal decompensation in 10 (6.2%), and hypotony (intraocular pressure equal to or less than 5 mmHg) in 9 (5.6%). A chi-square analysis of preoperative characteristics showed that hypotony (P = 0.007), macular detachment (P = 0.020), a history of cataract extraction (P = 0.003), poor visual acuity (P = 0.000), giant tear extent greater than 180 degrees (P = 0.004), and higher grade proliferative vitreoretinopathy (P = 0.000) all predicted a poor visual outcome. Vitreon (Vitrophage, Inc., Lyons, IL) was left in 16 eyes (9.9%) for an extended postoperative retinal tamponade for between 3 and 1034 days (mean, 87.2 days). The Vitreon was well tolerated, and these eyes experienced a similar outcome and rate of retinal reattachment to the rest of the group. CONCLUSIONS: Vitreon is a safe and useful adjunct to pars plana vitrectomy in the management of giant retinal tears and may, additionally, be the perfluorocarbon liquid that can be used most safely as a temporary postoperative tool for extended retinal tamponade, reinforcing its role as a useful adjunct in the management of these complex retinal detachments. PMID- 9224471 TI - Post-traumatic proliferative vitreoretinopathy. The epidemiologic profile, onset, risk factors, and visual outcome. AB - PURPOSE: The purpose of the study was to characterize the clinical development of proliferative vitreoretinopathy (PVR) after trauma in the human eye. METHODS: A chart review was performed on the records of 1564 patients with ocular trauma seen at a large metropolitan hospital. The frequency, type of ocular trauma, time to onset, potential risk factors, and visual outcome for PVR were evaluated. RESULTS: Proliferative vitreoretinopathy occurred in 71 (4%) of 1654 injured eyes. Of these 71 injured eyes, 30 (42%) resulted from rupture, 15 (21%) from penetration, 13 (18%) from perforation, and 7 (10%) from confusion. Six (9%) were associated with an intraocular foreign body (IOFB). The frequency of PVR following perforation, rupture, penetration, IOFB, and contusion was 43%, 21%, 15%, 11%, and 1%, respectively. Overall, those eyes that developed PVR had a poorer visual outcome, with PVR being the primary reason for visual loss. The time from injury to onset of PVR was shortest after perforation (median, 1.3 months), followed by rupture (2.1 months), IOFB (3.1 months), penetration (3.2 months), and contusion (5.7 months). Vitreous hemorrhage was the strongest independent predictive factor for the development of PVR. A long, posteriorly located wound and persistent intraocular inflammation were also important risk factors for PVR. CONCLUSIONS: These results suggest that PVR is a common complication following a variety of ocular injuries, and that it is associated with a poor visual outcome. Its frequency, onset, and outcome are strongly dependent on the nature of the trauma. Specific high-risk groups are identified as candidates for more aggressive therapy. PMID- 9224472 TI - Application of polymerase chain reaction assay in the diagnosis of orbital granuloma complicating atypical oculoglandular cat scratch disease. AB - BACKGROUND: Parinaud oculoglandular syndrome is uncommon. Most cases are caused by cat scratch disease (CSD), recently discovered to be associated with the pathogen Bartonella henselae. Before isolation of the micro-organism, diagnosis relied on the presence of characteristic clinical features. However, atypical cases could cause diagnostic problems. With the development of an indirect fluorescent antibody test and polymerase chain reaction (PCR) assay, oculoglandular CSD can be diagnosed readily. METHODS: The authors report a case of atypical Parinaud oculoglandular syndrome in a 51-year-old woman who presented with an inferior conjunctival forniceal mass extending into anterior orbital tissues. Blood and operative tissue specimens were obtained for routine screening and histopathologic analysis but more specifically for serologic analysis, culture, and PCR assay for B. henselae. Computed tomography was performed to delineate the mass. RESULTS: Cultures for B. henselae were negative. Initial serologic analysis demonstrated a low IgG response without detectable IgM, but 1 month later had undergone a fourfold rise in IgG, again without detectable IgM. Histopathologic analysis showed a nonspecific necrotizing granulomatous inflammation consistent with but not diagnostic of CSD. Polymerase chain reaction assay for B. henselae was strongly positive. Computed tomographic scan showed a preseptal and anterior orbital inflammatory process. CONCLUSIONS: Cat scratch disease due to B. henselae should be suspected in patients with atypical conjunctival inflammation associated with regional lymphadenopathy. PCR assay is extremely useful in establishing the diagnosis. The PCR assay offers the additional advantage of early diagnosis because the test is positive early in the disease. Antibiotic therapy remains controversial. In this case, surgical excision hastened resolution of the conjunctival inflammation. However, the lymphadenopathy responded poorly to antibiotics. PMID- 9224473 TI - Laser microsurgery for superficial T1-T2 basal cell carcinoma of the eyelid margins. AB - BACKGROUND: Basal cell carcinoma (BCC), the most common malignancy of the eyelid margins, poses therapeutic problems. Surgery, radiation therapy, and cryotherapy are the currently accepted methods for the treatment of this affliction. To verify the technical and clinical effectiveness of the surgical laser method, a specific approach was developed by performing laser-combined procedures under microscopic control. METHODS: A series of 26 patients underwent carbon dioxide (CO2) laser microsurgical excision of 27 primary superficial BCCs of the eyelid margins. Eighteen tumors were T1 and 9 were T2. The lesions were located at the lid margins in 18 and at the canthus in 9 cases. The eyelash line was involved in all cases, whereas intermarginal space was involved in 17 cases, without extension to the conjunctival border. Six lesions were in the lacrimal region. Median linear extent of the lesion was 5 mm (range, 4-10 mm). Treatment was performed with the patient under local anesthesia in a Day Hospital regimen. The authors used the microscope-mounted CO2 laser as a scalpel to excise the tumor mass, thus obtaining the specimen for histologic evaluation. The authors treated the deep and lateral resection margins with laser vaporization and left the wound bed to heal by secondary intention. RESULTS: No significant complications were observed. As full-thickness eyelid resections were avoided, the authors noted conservation of lid function and cosmetic aspect in all patients. With a median follow-up of 73 months (range, 18-118), only one patient had tumor recurrence after 22 months. This tumor, located at the outer canthus, had a second microsurgical laser excision, and the patient is disease free 51 months after the last treatment. CONCLUSIONS: Laser microsurgery appears to be a safe and effective treatment method for primary superficial T1 and T2 BCC of the eyelid margins without conjunctival extension. PMID- 9224474 TI - Granulomatous blepharitis as a sign of Melkersson-Rosenthal syndrome. AB - OBJECTIVE: The objective of our study was to describe the ophthalmic features and histologic eyelid findings of Melkersson-Rosenthal syndrome. DESIGN: Three patients with eyelid edema underwent eyelid biopsy to establish the diagnosis of Melkersson-Rosenthal syndrome. RESULTS: Of the three patients, only one patient manifest the classic triad of facial edema, facial paralysis, and furrowed tongue. Histoloppgically, the eyelid skin in all patients showed characteristic perilymphatic granulomas with marked dermal edema. CONCLUSIONS: The histopathologic features assist in establishing the diagnosis of Melkersson Rosenthal syndrome in oligo- and monosymptomatic patients. Melkersson-Rosenthal syndrome should be considered in patients presenting with eyelid edema of unknown etiology and biopsy performed. PMID- 9224475 TI - Transcanalicular neodymium: YAG laser for revision of dacryocystorhinostomy. AB - BACKGROUND: Laser-assisted dacryocystorhinostomy (DCR) has failed to match the success rates of external DCR. It has been suggested that this technology may be best suited for revision of failed DCR cases. The authors prospectively evaluated the efficacy of transcanalicular laser-assisted revision DCR (TCLARDCR). METHODS: A neodymium:YAG (Nd:YAG) laser was used for transcanalicular revision of 24 failed DCRs. Failure had followed one (n = 15), two (n = 7), or three (n = 2) previous external DCRs. RESULTS: Mean duration of the surgery was 78.2 minutes. Success was achieved in 11 cases (46%; mean follow-up, 20 months). There was no correlation between early loss of stents and failure. Three cases had partial relief of symptoms. Three of the failures unsuccessfully underwent further TCLARDCR. CONCLUSIONS: The authors' success rate of 46% with TCLARDCR compares poorly with the 85% success for external revision DCR. With TCLARDCR, specific anomalies like the sump syndrome cannot be addressed adequately. There is a theoretical risk of canalicular injury. Laser lacrimal surgery also is equipment dependent and more costly than external DCR. The TCLARDCR cannot be recommended for revision DCR using the Nd:YAG laser (Lasersonics, Milpitas, CA). PMID- 9224476 TI - Multidisciplinary management of refractory orbital rhabdomyosarcoma. AB - PURPOSE: Combined chemotherapy and radiation therapy have improved the survival of children with primary orbital rhabdomyosarcoma, but recurrence or persistence of the local orbital tumor still occurs. There are no established guidelines for dealing with these uncommon patients, and the authors present a review of the combined method treatment and outcome of children with refractory primary orbital rhabdomyosarcoma. METHODS: From clinical databases, 67 children with orbital rhabdomyosarcoma were identified. Seven (10%) of the 67 children had tumors refractory to combined chemotherapy and radiation therapy and underwent exenteration or eye-sparing tumor excision. Their clinical course and outcome were reviewed retrospectively. RESULTS: No patient was lost to follow-up, which ranged from 3.2 to 11 years. Five (71%) of the seven children with refractory tumor are still alive at more than 3 years after surgery (3.2-11 years; mean, 6.9). In one of the two children who died, tumor extended beyond the operative margins at exenteration, and the other child died with regional metastasis within a month of exenteration. CONCLUSIONS: Although more than 90% of children with orbital rhabdomyosarcoma respond to combined therapy by pediatric oncologists and radiotherapists, local orbital (salvage) surgery by ophthalmologists may be of value in the minority of children with refractory tumors. All of the five surviving children appear to be disease free. PMID- 9224477 TI - A flood of rewards. Volunteers set an example for all of us. PMID- 9224478 TI - Document imaging in medicine. How long can you do without it? PMID- 9224479 TI - Is routine screening for colorectal cancer justifiable? These gastroenterologists say YES! AB - While methods of screening for colorectal cancer undoubtedly will be refined and new techniques developed, there is ample evidence to support use of the currently employed protocol: annual fecal occult blood testing and periodic flexible sigmoidoscopy. Aggressive attempts to educate physicians and patients on the importance of such screening are needed. Primary care physicians can play an important role in ensuring patient compliance and reducing the incidence of this serious public health problem. PMID- 9224480 TI - Pitfalls in diagnosis of Lyme disease. What you need to know about serologic testing. AB - Lyme disease can be difficult to recognize because not all patients have erythema migrans or other classic symptoms. Therefore, laboratory testing has become an important aid to clinical diagnosis. But the story doesn't end there-serologic testing presents another set of problems and concerns. Drs. Still and Ryan explain how aspects of the disease itself and various factors inherent in the available tests can affect laboratory results. PMID- 9224481 TI - To screen or not to screen for HIV in pregnant women. Pros and cons of routine screening and use of home test kits. AB - Emotions run high when it comes to who should and who should not be tested for HIV. Now, as the debate heats up over screening of all pregnant women and their babies, primary care physicians may find themselves caught in the fray. Recent approval of home HIV test kits allows patients to test themselves, but can the results be trusted? Dr. Wong summarizes the contentious issues, looks at the debate from both sides, provides supportive documentation when available, and points out the "knowledge gaps." PMID- 9224482 TI - Epilepsy management. Issues in medical and surgical treatment. AB - After a single seizure, about 40% of patients have recurrence. The main features correlating with recurrence are cause, seizure type, EEG findings, family history of seizures and, possibly, the presence of a prior febrile seizure, Todd's paresis, and other abnormal neurologic findings. A number of medications are available for treatment. Withdrawal from medication is successful in 60% to 70% of patients. Several factors favor successful drug taper. These include a seizure free status for at least 2 years during drug therapy, a single type of seizure (partial or generalized), young age at seizure onset, and an epilepsy syndrome with a tendency to remit. Surgery can be considered in certain patients with surgically remediable syndromes. Candidates typically have seizures that impair consciousness, that cause falling with injury, that have adverse psychosocial or social effects, and that persist after trials of three appropriate medications. A multidisciplinary evaluation should take place at a surgery center with experience and documented success. Favorable results from surgery can be expected in a large proportion of patients. PMID- 9224483 TI - Women's issues in epilepsy. Menses, childbearing, and more. AB - A variety of special issues arise in caring for women with epilepsy. Seizures related to the menstrual cycle (catamenial epilepsy) can be controlled by several treatment strategies. Oral contraceptives are not contraindicated, but higher dose agents are needed if enzyme-inducing antiepileptic medications are given concomitantly. Sexual and reproductive dysfunctions are common but often go unrecognized. In the past, many people with epilepsy were wrongly advised against reproduction. With appropriate management, most women with epilepsy deliver healthy children. A better understanding of these concerns will allow physicians to improve the lives of women with epilepsy. PMID- 9224484 TI - New antiepileptic drugs. Overcoming the limitations of traditional therapy. AB - Several new antiepileptic drugs have become available recently. Since seizures and epilepsy are common, primary care physicians are likely to encounter a patient who is taking one of these new medications. Successful medical management of epilepsy requires a proper understanding of medication half-life, indications, and side effects. Felbamate has a broad spectrum of efficacy but is limited by side effects and idiosyncratic reactions. Fosphenytoin has the efficacy of phenytoin and offers the advantage of intramuscular and intravenous dosing without the significant adverse effects associated with intravenous phenytoin; however, it is expensive. Gabapentin has minimal side effects and drug interactions yet has limited efficacy for seizures. Lamotrigine has broad seizure efficacy but requires a slow adjustment to therapeutic levels. Topiramate has minimal drug interactions, but therapy must be initiated slowly to avoid side effects. All of the new antiepileptics hold great promise in the management of patients with recurrent seizures. PMID- 9224485 TI - Seizures in special populations. Children, the elderly, and patients with coexistent medical illness. AB - Epilepsy management may need to be modified in certain patient groups. The management of epilepsy in all age-groups, and particularly in children, may be best approached from a disease-based model that recognizes specific epilepsy syndromes. Accurate diagnosis of pediatric epilepsy syndromes allows the physician to improve medication selection, estimate duration of treatment, and counsel patients and family members about aggravating factors and prognosis. Epilepsy management in elderly patients requires knowledge of age-related changes in the metabolism and protein binding of anticonvulsants and how these changes affect use of particular antiepileptic drugs. Concurrent medical illness, especially renal and hepatic disease, may alter drug distribution, metabolism, and excretion in some patients. In addition, a number of medications used to treat coexistent medical or psychiatric illness are associated with seizure provocation. Knowledge of these factors can improve epilepsy management in special patient populations. PMID- 9224486 TI - Varicella-zoster virus infection. The complex prevention-treatment picture. AB - The highly prevalent and contagious varicella-zoster virus is usually benign in healthy persons but may cause substantial morbidity in immunocompromised patients and some adults. New developments in prevention and treatment, as discussed in this article, offer attractive options but also present difficult management decisions for primary care physicians. PMID- 9224487 TI - Overpowering pain. A serious problem comes out of the closet. PMID- 9224488 TI - Lipid screening in children and adolescents. Identifying patients with familial hypercholesterolemia. Institute for Clinical Systems Integration. PMID- 9224489 TI - Ultrastructural features of the pineal gland from cold-exposed golden hamsters. PMID- 9224490 TI - The nervous system of ctenophores. III. Ultrastructure of synapses. PMID- 9224491 TI - Death of the central neuron: an electron microscopic study of thalamic retrograde degeneration following cortical ablation. PMID- 9224492 TI - Extracellular space and membrane changes in brain owing to different alkali metal buffers. PMID- 9224493 TI - Freeze-etching images of central myelinated nerve fibres. PMID- 9224494 TI - Quantitative electron microscopy on the injured hypoglossal nucleus in the rat. PMID- 9224495 TI - The sensory hairs and tectorial membrane of the basilar papilla in the lizard Calotes versicolor. PMID- 9224496 TI - Active cell death (apoptosis) in liver biology and disease. PMID- 9224497 TI - Regulation of liver-specific gene expression. PMID- 9224498 TI - Control of vesicle traffic in hepatocytes. PMID- 9224499 TI - Regulation and function of the multidrug resistance genes in liver. AB - The P-glycoproteins are integral membrane proteins that function as ATP-dependent transporters. The multidrug resistance genes which encode P-gp comprise a small gene family, with 2 members in humans and 3 in rodents. The P-gp encoded by the mdr1 gene functions as a drug efflux pump to remove drugs from cells and may serve as a barrier to protect cells from cytotoxic agents. In normal tissues, P gp is localized on the luminal surface of transporting epithelia in the liver, kidney, small intestine, testes, and blood-brain barrier. Transient exposure to drugs transcriptionally increases the level of expression of the mdr1 genes, however, the cellular pathways critical to this regulation are yet unknown. This observation may have some implications on the level of expression in tumors and response to chemotherapy. Examination of the basal level of MDR expression in tumors may not be a reliable predictor of the effect of P-gp on chemotherapy. Induction of MDR transcription by drugs may further impede the effectiveness of anti-cancer agents. This is most obvious for drugs which are substrates for P-gp transport, however, it also applies significantly to compounds which are not themselves substrates but affect the response to other drugs simultaneously or subsequently administered. A clear understanding of the mechanisms that regulate basal and drug-induced mdr transcription will facilitate development of novel agents which circumvent this obstacle or permit targeted modification of mdr expression. Expressed on the bile canalicular surface of the liver, P-gps represent the first ATP-dependent biliary transporters to be characterized. The P gp encoded by mdr2 is the major form of P-gp expressed in normal liver and transports phospholipids into bile. A defect in this protein leads to severe liver disease caused by chronic inflammation of the biliary system that results from high concentrations of free bile salts. The cellular origin and molecular basis of the ensuing liver tumors in these mice are unclear. It is possible that the chronic damage to the biliary ductules causes an increased growth rate of the surrounding cells, including putative stem cells in the liver. Thus, these mice may serve as a model for carcinogenesis in which the liver is under constant promotion placing the proliferating cells at increased risk to further genetic alterations or expansion of preexisting, but normally quiescent, mutations. Mdr2 deficient animals may also provide a model for human chronic inflammatory liver disease. Clearly, these exciting results indicate that further characterization of the P-gps as normal physiologic canalicular membrane transporters is necessary. PMID- 9224501 TI - Ethanol oxidizing enzymes: roles in alcohol metabolism and alcoholic liver disease. AB - The elucidation of nucleotide and amino acid sequences of the genes and enzymes involved in the metabolism of ethanol has led to the ability to genotype individuals simply and rapidly. Although the different isozymes were once thought to be a likely explanation for between individual differences in alcohol elimination rates, this has not been found to be the case. Other explanations that remain to be investigated include potential regulatory variants in the genes that alter the level of expression of the enzymes, and genetic influences on activity of the malateaspartate shuttle and rates of mitochondrial NADH reoxidation. However, the isozymes encoded by ADH2*2 and ALDH2*2 have been found to influence alcohol drinking behavior or alcoholism substantially. This supports the original premise that the metabolic disposition of ethanol affects individuals' responses to it. The results suggest that any additional variants might also contribute to the spectrum of individual drinking preferences. Among heavy drinkers, the influence of the isozymes on risk of alcoholic liver disease has not been found to be great, suggesting that many other factors, perhaps interacting with the enzyme polymorphisms, are involved in determining susceptibility. PMID- 9224500 TI - Molecular biology of bilirubin metabolism. AB - As the genes encoding the glucuronidating enzymes are discovered, it is evident that glucuronidation is a magnificent example of how in evolution, man became adapted to his "intoxicating" environment. A superfamily of genes is necessary to dispose of the toxins and carcinogens that are encountered by inhalation and ingestion. The enzymes that glucuronidate endogenous compounds are members of this large family. For the clinician, it is important to remember that jaundice may sometimes be the result of interactions at the level of bilirubin glucuronidation. When jaundice results from inactivation of members of the UGT1 family, conjugation of certain phenols, such as the anesthetic propofol, or synthetic estrogens, such as ethinylestradiol, can also be impaired. In the case of severe bilirubin glucuronidation deficiencies, such as the Crigler Najjar syndrome type I, there are exciting prospects for a possible cure by gene therapy. PMID- 9224502 TI - Hepatic lipoprotein metabolism: recent molecular insights. PMID- 9224503 TI - Gene therapy for human liver disease. AB - Investigators working in the area of gene therapy believe the potential for advances in all medical disciplines is enormous. It is humbling, however to appreciate how far we need to go, as the field is truly in its infancy. Gene transfer technologies currently under evaluation in clinical trials have major limitations. Vector systems used in the clinics by the year 2000 probably have yet to be discovered. An additional lesson learned is that efforts at gene therapy are hampered by a lack of knowledge of the basic biology of the target organ and pathogenesis of the underlying disease. Successful gene therapy programs will critically evaluate the field and through fundamental research move steadily forward toward the long-term goal of truly effective therapy for a wide spectrum of disease. In the near future, two liver diseases are the most likely to be treated with gene therapy. The evaluation of patients with familial hypercholesterolemia is ongoing, and once approved, more candidates will be enrolled for therapy. Progress also has been made in creating vectors for the treatment of ornithine transcarbamylase deficiency. It remains to be seen whether adenoviruses or retroviruses will be used first in attempts to control this disease. Although the inflammatory response noted with current recombinant viruses is a formidable problem, the efficiency of gene transfer into the liver with these vectors makes continued study worthwhile. PMID- 9224504 TI - Use of novel beta-L(-)-nucleoside analogues for treatment and prevention of chronic hepatitis B virus infection and hepatocellular carcinoma. PMID- 9224505 TI - Autoimmune disorders associated with hepatitis C. PMID- 9224507 TI - Urea cycle disorders: clinical paradigm of hyperammonemic encephalopathy. PMID- 9224506 TI - Pharmacological treatment of portal hypertension. PMID- 9224508 TI - Pathogenesis of brain edema in fulminant hepatic failure. PMID- 9224509 TI - Artificial hepatic support systems. AB - Severe acute liver failure is associated with high mortality. Improved respiratory and hemodynamic management together with intracranial pressure monitoring and aggressive treatment of cerebral edema have greatly improved patient care. However, many patients die despite optimal medical treatment, because of failure to arrest the progression of cerebral edema. This in turn result in brain stem herniation with rapid neurologic deterioration and death. Liver transplantation has emerged as the definitive treatment for patients with severe acute liver failure. Unfortunately approximately up to one half of the patients with this severe form of liver failure will die while awaiting liver transplantation. There is thus a clear need for a liver support system to provide a "bridge" to transplantation. Over the years, many "bridge" systems were introduced which promised effective support but had no wide clinical success. Because of our incomplete understanding of the pathophysiology of liver failure and development of cerebral edema, it was felt that use of isolated hepatocytes or ex vivo whole liver perfusion would provide both detoxifying and synthetic functions. Whole liver perfusion appears to be effective but cumbersome and costly because it would require each center, where patients are being treated, to maintain animal colonies for patient treatment. Therefore, cryopreserved isolated xenogeneic hepatocytes appear to be the best candidates for building a "bridge" system. In a preliminary clinical study, we have used a porcine hepatocyte-based liver support system (Bioartificial Liver: BAL) to treat patients with acute liver failure as well as patients with acute exacerbation of chronic liver disease. Patients in the first group, who were candidates for transplantation, were successfully bridged to a transplant with excellent survival. No obvious benefit from BAL treatments was seen in the second group. In this group patients where cerebral edema is not a major component of the clinical presentation, it is possible that long-term support will be needed with repeated treatments over several weeks to provide adequate synthetic and detoxifying liver function until the patients' livers recover. For such liver recovery to take place, these chronic patients will need to be treated earlier in the course of their disease when they still have some residual liver mass as well as regenerative capacity. Prospective controlled trials will be initiated as soon as the current phase I study is concluded in order to determine the efficacy of this system in both patient populations. PMID- 9224510 TI - Laparoscopic hepatobiliary surgery. AB - Laparoscopic hepatobiliary surgery has advanced rapidly in the past few years. Laparoscopic cholecystectomy has been the most significant advance and its value will become even greater as the incidence of biliary injuries is decreased. LCBDE and staging laparoscopy are very promising procedures. Other hepatobiliary laparoscopic operations such as liver resections and biliary-enteric anastomoses are still in developmental stages and will require a longer period of assessment. PMID- 9224511 TI - Immunosuppressive therapy in liver transplantation: principles and practice. PMID- 9224513 TI - Laparoscopic appendicectomy: pros & cons--literature review of 4190 cases. AB - The aim of this literature review is to evaluate current data on laparoscopic and open appendicectomy in order to establish a new gold standard in surgery. The authors have reviewed 4190 appendicectomies, 3322 laparoscopic appendicectomies (LA) and 868 open appendicectomies (OA), from 22 studies published between February 1992 and June 1994. Four studies dealt exclusively with pediatric patients (n = 1558). The analysis compares surgical technique, operating time, pathological findings, major and minor complications, postoperative pain and costs. The strongest arguments against LA are the increased rate of major complications, the increase of overall cost and the negative effect of the learning curve. The argument in favour of LA are the significant reduction of minor complications, the shortening of the postoperative hospitalization and of the time to resume full activity. The benefit of LA regarding the amount of analgesia requirement and the time needed to resume normal feeding after surgery is still controversial. The authors have reached the conclusion that LA might emerge as the first choice for appendicectomy. More studies are necessary to confirm this trend. PMID- 9224512 TI - Prophylaxis of venous thromboembolism in surgery. PMID- 9224514 TI - Prognosis of gastric carcinoma with emphasis on lymph node status. AB - A retrospective study of surgical treatment for 123 patients with primary gastric carcinoma was made. Tumour-stage, presence of distant organ metastases, lymph node involvement, depth of wall-penetration and curative resection were the important prognostic criteria. Five-year survival rate was 22.8% in the whole series and 37.5% after curative (R0 M0) resection (p = 0.0029). Seventy-two patients (58.5%), i.e. 47 without (N0) and 25 with lymph node metastases (N1 + N2), underwent, R0-resection. The overall recurrence rate was significantly lower (4% vs. 36%; p = 0.004) and the 5-year survival rate higher (46% vs. 28%; p = 0.007) for the R0-N0-group than for the R0-N1 + N2-group. Distant organ metastases-related mortality was similar in these two groups of patients (51% vs. 60%; p = 0.7). CONCLUSION: The poor prognosis of gastric carcinoma indicates the need for improved surgical technique and/or adjuvant therapy, but the necessity for adjuvant therapy without having performed D2 gastrectomy does not seem founded. PMID- 9224515 TI - An intra-operative technical aid to miniplating of mandibular fractures by fracture segment apposition. AB - Internal plate osteosynthesis and lag-screw osteosynthesis are currently the preferred methods of fixation in the treatment of mandibular fractures. Rigid internal fixation (RIF) provides for functionally stable immobilization of the segments, avoiding the need for postoperative intermaxillary fixation (IMF) by "dental wiring" in the majority of cases. Reduction and fixation can be provided by so-called compression plates (type AO/ASIF). Non compressive mini-plates are more extensively used, however, because their size and malleability facilitate their transoral application. Peroperative problems may arise with the anatomical reduction of the fragments. We present a simple fracture reduction-compression technique that can be used in combination with either compression and non compression mini-plate fixation. Its use can be extended to reduction and fixation in selected areas of difficult surgical access. PMID- 9224516 TI - The laparoscopic resection of a benign stromal tumour of the duodenum. AB - A 72-year-old-female presented intermittent retrosternal pain, heartburn and dysphagia. Computerized CT-Scan showed a large mass with a cross-sectional diameter of 5 cm at the lateral side of part II of the duodenum. The preoperative histology was unclear. The tumour was successfully removed by laparoscopic approach. PMID- 9224517 TI - Conservative surgical treatment for a giant thoracoabdominal benign teratoma. AB - The authors describe an adult patient with a giant, cystic teratoma (33 x 22 x 18 cm) involving the posterior mediastinum and retroperitoneum. A conservative surgical approach consisting of a partial endocystectomy plus injections of tetracycline into the residual cavity, was performed. Two months after surgery, a CT scan showed a 6 x 5 x 3 cm residual cyst. The authors believe that the excision of the inner surface of the cystic lesion and the intracystic administration of tetracycline, may successfully prevent the accumulation of fluid which is the main cause of the progressive enlargement of such benign cystic teratomas. PMID- 9224518 TI - Uretero-arterial fistula: two observations. AB - Two cases of life-threatening haematuria, secondary to an uretero-arterial fistula, are reported. Both cases present predisposing causative factors. One patient had a combination of previous aorto-bifemoral bypass grafting, an iliac artery aneurysm (retrogradely perfused), and an indwelling ureteral stent for ureteral compression. The other patient had previous aortoiliac surgery and obstructive uropathy with chronic urinary tract infection. Preoperative diagnosis of uretero-arterial fistula was made in only one patient. He was successfully operated (exclusion of the iliac aneurysm). In the other patient, nephrectomy was attempted to control reno-ureteral bleeding of unknown origin. Fatal recidive of brisk haematuria occurred some days later. Factors contributing to the development of uretero-arterial fistula, their diagnosis and optimal treatment are discussed. PMID- 9224519 TI - Intra-arterial thrombolysis followed by elective surgery for thrombo-embolic popliteal aneurysms. AB - Two patients with acute limb-threatening lower extremity ischaemia as a result of a thrombosed and embolizing popliteal artery aneurysm are described. Both patients were successfully treated with intra-arterial thrombolysis and subsequent elective vascular reconstruction. Thrombolysis might be an effective method to identify the underlying cause of limb ischaemia and to recanalize the run-off vessels leading to better bypass patency rates. However, this management is only indicated in selected cases of acute limb ischaemia without motor or sensory deficit. PMID- 9224520 TI - Successful management of acute aortic dissection in a heart transplant recipient. AB - A case of type III aortic dissection which occurred fourteen months after heart transplantation is presented. Medical therapy was instituted to achieve controlled hypotension. The evolution was favorable and the patient could be discharged after one month. Hypertension and increased ejection fraction after transplantation could have been predisposing factors via an increase of the shear stress in the aorta. PMID- 9224521 TI - Posttraumatic pneumomediastinum: not always cause for alarm. AB - A case of pneumomediastinum following a minor blunt thoracic trauma is presented. As no underlying organic lesion could be found, the pathophysiology must have been that of a spontaneous pneumomediastinum, i.e. alveolar rupture with subsequent dissection of air along the bronchovascular sheats to the mediastinum. The pathophysiology, clinical presentation, diagnosis and treatment of the spontaneous pneumomediastinum are discussed. With regard to the treatment it seems most important not to be too aggressive, because spontaneous pneumomediastinum is a benign and self-limiting condition. Spontaneous regression within a week can be expected. PMID- 9224522 TI - Wilms' tumour presenting as a pulmonary embolism. AB - Wilms' tumour (nephroblastoma) is one of the most common solid malignant tumours in infancy and childhood. In about 40% of cases, invasion of the renal vein is present. Rarely, these tumours extend into the inferior vena cava and right atrium. We discuss a 7-week-old girl who presented with acute massive pulmonary embolism, and was found to have a large tumour of the left kidney. Later on, the diagnosis of Wilms' tumour was confirmed. Wilms' tumour presenting as massive pulmonary embolism is extremely rare; to our knowledge this is the fourth case described in literature. We review the cases previously described, and comment on the diagnostic and therapeutic features of this clinical entity. PMID- 9224523 TI - Neuritic plaque evolution in Alzheimer's disease is accompanied by transition of activated microglia from primed to enlarged to phagocytic forms. AB - Activated microglia, overexpressing the potent neuroactive cytokine interleukin 1, have been implicated as a driving force in the evolution of diffuse amyloid deposits into diagnostic neuritic plaques in Alzheimer's disease. To evaluate this role further, we used double-label immunohistochemistry to classify and quantify plaque-associated and non-plaque-associated activated interleukin-1 immunoreactive microglia in parahippocampal tissue from 11 patients with Alzheimer's disease. These activated microglia were subclassified as primed (only slightly enlarged), enlarged, or phagocytic (enlarged with heterogeneous cytoplasmic contents). We further determined the distribution of these microglial subtypes among four defined plaque types. Most (84%) primed microglia were not plaque associated, although 13% were present in diffuse non-neuritic plaques and 3% were present in diffuse neuritic plaques. In contrast, most enlarged (55%) and phagocytic (91%) microglia were plaque associated. Of plaque-associated enlarged microglia, most (71%) were found in diffuse neuritic plaques with the remainder evenly distributed between diffuse non-neuritic and dense-core neuritic plaques (15% each). Of plaque-associated phagocytic microglia, a few were present in diffuse non-neuritic plaques (5%), but most were found in diffuse neuritic plaques (62%) and dense-core neuritic plaques (33%). These findings show preferential association of primed microglia with diffuse amyloid deposits and imply that microglial transformation from primed, to enlarged, to phagocytic types occurs in concert with the evolution of amyloid plaques from diffuse amyloid deposits to the neuritic beta-amyloid plaque forms in Alzheimer's disease. Microglial phagocytic activity in neuritic plaques may reflect involvement in the processing of diffuse amyloid into condensed beta-amyloid, or in clearance of neuritic debris. PMID- 9224524 TI - Cell cycle markers in the hippocampus in Alzheimer's disease. AB - Using immunohistochemistry we have analysed the nuclear expression of cyclins A, B, D, and E in neurones in the hippocampi of control subjects and patients suffering from various neurodegenerative disorders including. Alzheimer's disease (AD). Cyclins A and D could not be detected but varying degrees of cyclin E expression were found in all patient groups including control subjects. Cyclin B expression was not detected in control subjects but it was expressed in the subiculum, dentate gyrus and CA1 region in patients with AD-type pathology and in the CA2 region and the dentate gyrus of cases of Pick's disease. These results suggest that some neurones may have re-entered the cell cycle. The expression of cyclin E without cyclin A expression may indicate an arrest in G1 with the possibility of re-differentiation and exit from G1 to G0. The expression pattern of cyclin E indicates that re-entry into the cell cycle is possible even in control patients, but it is accentuated in patients with AD-related pathology. However, cyclin B was only expressed in AD patients and occurred in areas that were severely affected by pathology. Neurones with cyclin B-reactive nuclei in AD were AT8 positive but did not contain fully developed tangles. In neurones, where cyclin B is expressed, it would appear that the G1/S checkpoint has been bypassed and that the cell cycle is arrested in G2. It is proposed that these neurones do not have the opportunity for subsequent re-differentiation. Since factors known to be present in G2 seem to be responsible for microtubule destabilisation and hyperphosphorylation of tau we hypothesise that cell cycle disturbances may be important in the pathogenesis of AD. PMID- 9224525 TI - Up-regulation of intercellular adhesion molecule 1 in cerebral microvessels after cortical contusion trauma in a rat model. AB - A study was made on the expression of the intercellular adhesion molecule 1 (ICAM 1) in cerebral microvessels after cortical contusion trauma of the rat brain. The trauma was produced by a free-falling weight on the exposed dura of one fronto parietal lobe. Immunohistochemistry was done on cryostat sections using a monoclonal antibody and the reaction product was visualized using the avidin biotin-peroxidase complex method. Control and sham-operated rats showed immunostaining of some penetrating arteries of the cerebral cortex, the epithelial cells of the choroid plexus and occasional microvessels of the brain parenchyma. The same pattern of immunostaining was seen in rats that were subjected to trauma and killed after 30 min. All rats with contusion trauma that were allowed to survive for 6-72 h showed a substantial increase in the number of immunostained capillaries throughout the site of the lesion. The ipsilateral hippocampus showed a mild to moderate increase in the number of immunostained microvascular profiles. This phenomenon was also present in the lateral thalamus of some rats. The staining was seen as an uninterrupted line at the position of the endothelial cells, indicating an upregulation of this adhesion molecule after brain trauma. Up-regulation of ICAM-1 is a well-known phenomenon in inflammatory and ischemic lesions of the brain but has not previously been described in detail in traumatic brain injury. ICAM-1 may be involved in the production of several post-traumatic events such as leukocyte adhesion, microcirculatory disturbances and edema formation. PMID- 9224526 TI - Accumulation of wild-type p53 in astrocytomas is associated with increased p21 expression. AB - Approximately one quarter of human astrocytomas show immunohistochemical positivity for p53 protein but lack p53 gene mutations, which could reflect either an accumulation of wild-type p53 protein or an inadequate sensitivity of mutation detection. Since wild-type p53 up-regulates p21 expression, increased p21 expression in those astrocytomas with p53 accumulation in the absence of mutations would argue that the protein was wild type in these tumors. We therefore compared p21 expression with p53 gene and protein status in 48 primary human astrocytomas. Single-strand conformation polymorphism analysis and direct sequencing of the p53 gene showed mutations in 11 tumors (22.9%), while immunohistochemistry revealed positive staining in 19 cases (39.6%). Those tumors with p53 immunopositivity in the absence of p53 mutation had significantly increased p21 expression when compared to either mutant p53 or p53-immunonegative cases. Neither p53 nor p21 status correlated with proliferation indices, as assessed by Ki-67 immunohistochemistry. These results support the hypotheses that functionally wild-type p53 accumulates in some astrocytomas, and that alternative cell cycle checkpoints (such as the p16 pathway) may be more important than p21 in regulating proliferation in astrocytomas. PMID- 9224527 TI - Concomitant deficiency of beta- and gamma-sarcoglycans in 20 alpha-sarcoglycan (adhalin)-deficient patients: immunohistochemical analysis and clinical aspects. AB - We have investigated the expression, using immunohistochemistry, of beta- and gamma-sarcoglycans in the muscles of 20 patients in whom previous screening had revealed a deficiency of alpha-sarcoglycan. alpha-, beta- and gamma-sarcoglycans were absent in 7 patients and variably reduced in 8 patients, in 2 of whom beta sarcoglycan was more reduced than the alpha- and gamma-proteins. In 5 other patients with variably reduced alpha- and beta-sarcoglycans, gamma-sarcoglycan was completely absent. In all patients the distribution of hyposthenia at disease onset was similar, and predominantly involved pelvic girdle muscles; however, the age at onset and rate of disease progression were highly variable. In severely compromised patients, the onset of disease was before 10 years of age and gamma sarcoglycan or all three sarcoglycans were absent from muscles. Immunohistochemical analysis of sarcoglycans should be part of routine screening for muscle dystrophies to identify patients with sarcoglycanopathy. Gene analysis is necessary to identify the primary defect; however, sarcoglycan immunohistochemistry may be useful for indicating which gene to investigate. Further biochemical characterization of the interactions between these proteins is required to fully elucidate their roles in causing severe, moderate or mild muscular dystrophy. PMID- 9224528 TI - Neuropathology of the dentate nucleus in developmental disorders. AB - The dentate nucleus was examined histologically and immunohistochemically in 47 cases of nonprogressive developmental disorders. Neuronal loss and/or atrophy was observed in 13 cases, while mild neuronal lesions, characterized by dendritic swelling and/or the appearance of eosinophilic materials around the neurons, were exhibited in 20 cases. The former change was accompanied by diffuse central nervous system involvement, and the etiology was perinatal hypoxic ischemic encephalopathy, acute encephalopathy, and meningoencephalitis in most cases. On the other hand, most of the patients with kernicterus showed the latter change. Immunohistochemically, the mild neuronal lesions mimicking grumose, degeneration, described in some neurodegenerative diseases, seemed to reflect the changes of Purkinje cell terminals. It is suggested that secondary structural alteration of the dentate neurons in the absence of severe atrophy can occur in nonprogressive developmental disorders. PMID- 9224529 TI - Immunocytochemical study of pituitary oncocytic adenomas. AB - Using immunocytochemistry we have analyzed 8 pituitary oncocytomas, 14 null cell adenomas, and 2 oncocytomas of the parotid gland (Warthin's tumor). The proportions of adenoma cells that are positive for mitochondrial protein (MP), cytochrome oxidase (COX), and manganese-superoxide dismutase (Mn-SOD) were significantly higher in pituitary oncocytomas than in null cell adenomas (MP P < 0.001, COX P < 0.001, Mn-SOD P < 0.05). In pituitary oncocytomas, MP-positive cells were distributed unevenly but in clusters or in islets admixed with some MP negative cells, and corresponded to COX-positive cells. In contrast, almost all of the oxyphilic epithelial cells of Warthin's tumor were positive for MP, COX, and Mn-SOD. On the other hand, both pituitary tumors displayed similar findings with regard to the proportion of adenoma cells immunoreactive for copper/zinc-SOD and adenohypophysial hormones, the Ki-67 (MIB-1) proliferating cell index, and the mean number of argyrophilic nucleolar organizer regions. It was confirmed that immunocytochemical identification of MP and COX is useful for distinguishing pituitary oncocytomas from null cell adenomas. Although it remains to be determined whether oncocytomas originate from oncocytic changes of tumor cells or from neoplastic transformation of oncocytic cells, it appears that tumorigenesis of pituitary oncocytomas differs from that of Warthin's tumor. PMID- 9224530 TI - Exertional rhabdomyolysis and exercise intolerance revealing dystrophinopathies. AB - Exercise intolerance associated with myalgias, muscle cramps or myoglobinuria may be associated with a dystrophinopathy. A search for abnormal dystrophin expression (using immunohistochemistry, immunoblot and DNA analysis) was carried out in a series of 15 patients. They were selected because they presented exercise intolerance, negative biochemical tests (lipid, glycogen and mitochondrial metabolism) and abnormal immunohistochemistry with at least one anti-dystrophin antibody (anti-Dys 1, rod domain; anti-Dys 2, C terminus; anti Dys 3, N terminus). Lack of anti-Dys 1 immunoreactivity was seen in three patients and abnormal immunoreactivity with all three anti-dystrophin antibodies in two. Immunoblot confirmed the dystrophinopathy in these five patients only, and multiplex polymerase chain reaction DNA analysis revealed a deletion in the dystrophin gene in two of these patients, affecting the proximal part of the rod domain in one and the distal part of this domain in the other. The clinical, biological and histopathological features of the five patients reported here, together with the previous cases reported in the literature, are described and reveal that exercise intolerance associated with dystrophinopathy displays characteristic clinical, biological and immunohistochemical features and defines a new dystrophinopathy phenotype. The absence of staining in the rod domain provides a secure diagnosis of this syndrome. Dystrophinopathy is one etiology of idiopathic myoglobinuria, requiring genetic counseling. PMID- 9224531 TI - Hereditary polioencephalomyelopathy of the Australian cattle dog. AB - A vacuolar degeneration affecting primarily the gray matter in the central nervous system (CNS) of young Australian Cattle Dogs is described. An initial presentation of seizures was followed by a progressive spastic tetraparesis. Grossly evident bilateral and symmetrical foci of malacia were in the nuclei of the cerebellum and brain stem and the gray matter of the spinal cord. Microscopically, vacuolation of glial cells, dilation of the myelin sheaths and reactive astrocytosis characterized mild CNS changes. More advanced lesions displayed progressive dissolution of the neuropil, prominent vacuolation of reactive astrocytes, numerous glial fibrillary acidic protein-positive coiled astrocytic processes, neuronal vacuolation and loss with relative sparing of large neurons. Ultrastructurally marked mitochondrial accumulation and swelling were seen in astrocytes. In the appendicular muscles, changes interpreted as long term denervation atrophy accompanied by widespread expression of the neonatal isoform of myosin were observed. The character of the neurological sings, the nature and the distribution of the lesions within the neuroaxis have not been reported in domestic animals. An inherited biochemical defect, possibly mitochondrial, is proposed as the cause. Selected conditions with a bilateral and symmetrical distribution affecting the gray matter of domestic animals are summarized. PMID- 9224532 TI - Ubiquitin-related cytoskeletal abnormality in frontotemporal dementia: immunohistochemical and immunoelectron microscope studies. AB - Although reports of dementia lacking the distinctive non-Alzheimer-type histopathology have been increasing, the concept is still far from clear. It has become apparent that this population shows neuropathological heterogeneity, and some recent reports have proposed a classification or criteria for these disease conditions. Of the reported cases, frontotemporal dementia (FTD) of motor neuron disease is unique in that the neurons of the hippocampus and entorhinal cortex have ubiquitin-related abnormalities. Recently, a new ubiquitin-related abnormality, characterized by ubiquitinated inclusions in the neurites, has been found in some FTD cases. Using immunoelectron microscopy with immunogold particles, we have found that in these two disease conditions ubiquitinated inclusions consist of abnormal filaments of 10-15 nm in diameter. Our results support the speculation that there is a close relationship between ubiquitin and abnormal filaments in these two types of FTD, indicating that cytoskeletal related disorders may underlie certain types of FTD. PMID- 9224533 TI - The long incubation period in rabies: delayed progression of infection in muscle at the site of exposure. AB - The striped skunk (Mephitis mephitis) is a host of rabies in large areas of Canada and the United States. In each of two experiments, equal numbers of skunks in two groups were inoculated intramuscularly with low doses of a field strain of rabies virus (street rabies virus). In each experiment, skunks in one group surviving to 2 months were killed at this time and selected tissues were used for examination by the polymerase chain reaction (PCR) method or by immunohistochemistry for rabies antigen. Results of detailed examinations using PCR technology (experiment 1) indicated that muscle at the inoculation site contained viral RNA at 2 months postinoculation, when other relevant tissues on the route of viral migration and early entrance into the central nervous system were negative. The cellular location of virus/antigen, as determined immunohistochemically in experiment 2, was striated muscle fibers and fibrocytes. Our results indicate a major role of muscle (tissue) infection at the inoculation site in the long incubation period of rabies in skunks. These and related findings will be useful in rabies control and, if applicable to other species, will be relevant in postexposure treatment. PMID- 9224534 TI - Scanning electron microscopical study of the neurofibrillary tangles of Alzheimer's disease. AB - Neurofibrillary tangles (NFTs) have been ultrastructurally studied by various methods, leading to several three-dimensional models of paired helical filaments (PHFs). In this study, we present the scanning electron microscopic findings of NFTs in an autopsy case of Alzheimer's disease and clarify the three-dimensional structures of NFTs. NFTs were clearly defined in freeze-cracked nerve cells and consisted of two types of filamentous structures, straight and helical filaments. Straight filaments measured from 20 to 25 nm in diameter and had a smooth surface. They were slightly bent but mostly straight with no constrictions. One type of straight filaments ran in a bundle in the same direction, another was intertwined to each other. Most of the helical profiles of filaments usually measured about 28 nm in diameter, with a distance of 100 nm between periodic constrictions. They seemed to consist of a pair of isodiametric filaments of 10 nm in diameter. In addition, two unusual types of helical filaments were occasionally observed. One comprised thick filaments of about 38 nm in diameter, with a distance of 100 nm between constrictions; these helical filaments appeared to consist of two or more strands. The other comprised thin helical filaments of about 20 nm in diameter and regularly constricted at an interval of 50 nm. All types of the helical filaments examined in this case were leotropic. This result supports a protofilament model of PHFs. Scanning electron microscopy using the freeze-cracked and maceration method is a useful and simple method for three dimensional observation of the filamentous structures in NFTs. PMID- 9224535 TI - Delayed spongiform leukoencephalopathy after heroin abuse. AB - Here we report the clinical and pathological findings in a 30-year-old drug addict in whom an intravenous injection of heroin led to reversible coma with respiratory depression and heart failure. On regaining consciousness, the patient was found to have rhabdomyolysis with renal failure requiring dialysis and peripheral neuropathy. Three weeks later his neurological condition suddenly deteriorated and delayed encephalopathy developed, leading to death 20 days later. The neuropathological study of the brain disclosed pale, spongy myelin with diffuse reactive astrogliosis and microglial proliferation, without hypoxic necrotic lesions. The cerebral and cerebellar cortices were unchanged. The absence of typical hypoxic lesions and the presence of spongiosis with massive astrocytosis distinguished this case from the previously reported cases of delayed leukoencephalopathy following severe hypoxia. An immunocytochemical study designed to exclude an underlying alteration of the metabolic oxidative pathway detected normal expression of the respiratory chain complexes IV, III and V. Despite the absence of an oxidative chain alteration in our patient, we cannot exclude the possibility that an individual predisposition played a pathogenetic role in this delayed leukoencephalopathy. PMID- 9224536 TI - Hamartoma of the triceps surae muscle. AB - A 9-year-old, otherwise healthy girl presented with a 5-year history of pain in her right calf with retarded growth and development of an equinus contracture of her right leg. Magnetic resonance imaging showed an irregular mass with heterogeneous enhancement after contrast in her right triceps surae muscles, especially the soleus. Histological studies of this triceps surae muscle tissue revealed a haphazard distribution of adipose and connective tissue, striated and smooth muscle cells, vessels and lymphoid follicles, as well as nerve bundles which, together, were considered components of a hamartoma. PMID- 9224537 TI - Cystic ganglioneurocytoma outside the ventricular region. AB - Recently cases of ganglioneurocytoma and cerebral neurocytoma, very rare variants of central neurocytoma, have been reported. The former is characterized by differentiation toward ganglion cells and the latter by extraventricular origin in the cerebrum, but their existence as distinct clinicopathological entities, is controversial. We report an unusual case of neurocytoma, which arose extraventricularly from the frontal lobe, formed a large cystic lesion and showed ganglioid differentiation, in a 11-year-old girl. Following subtotal tumor resection, she showed a satisfactory clinical course and no evidence of recurrence. This is a very rare case of central neurocytoma-like tumor outside the ventricular system and also of ganglioneurocytoma. This case may provide some insight into the tumorigenesis and widen the clinicopathological concept of neurocytoma. PMID- 9224538 TI - Neurofibrillary pathology in the human paraventricular and supraoptic nuclei. AB - Severe neurofibrillary changes were identified in the paraventricular and supraoptic nuclei of elderly individuals using markers for Alzheimer's disease related intraneuronal pathology. This neurofibrillary pathology is remarkable in that the magnocellular paraventricular and supraoptic nuclei are particularly resistant to Alzheimer's disease. Moreover, the changes were observed even in non demented controls, indicating that they develop independently of Alzheimer's disease. The alterations in the paraventricular and supraoptic nuclei were consistently accompanied by neurofibrillary changes in the mediobasal hypothalamus. PMID- 9224539 TI - EAACI provocation tests with allergens. Report prepared by the European Academy of Allergology and Clinical Immunology Subcommittee on provocation tests with allergens. PMID- 9224540 TI - Carotid body dopamine content and release by short-term hypoxia: effect of haloperidol and alpha methyl paratyrosine. AB - Dopamine (DA) is thought to modulate the transduction of the hypoxic stimulus by the glomus cell in the carotid body (CB). The hypothesis tested here is that presynaptic DA D2 receptors (D2's) located on the type 1 cell function as autoreceptors to control DA release and/or synthesis. The aim of the study was to compare the effects of blocking D2's with haloperidol and DA synthesis with alpha methyl paratyrosine (AMPT) on the in vitro carotid body DA response to hypoxia. 54 CB's sampled from adult rabbits were incubated for one hour in a surviving medium bubbled with either 100% O2 or 8% O2 Sixteen CB's served as control (100% O2: n = 8, 8% O2: n = 8), 18 (100% O2: n = 8, 8% O2: n = 10) were sampled from rabbits pretreated with AMPT and 20 (100% O2: n = 12, 8% O2: n = 8) were incubated with micromolar concentrations of haloperidol. At the end of exposure. DA contained in the carotid body (DACB) and released in the surviving medium (DAr) were measured by HPLC. In 100% O2 DACB was not different between either AMPT or haloperidol and control, but DAr was significantly higher in the haloperidol group compared with control (mean +/- SE: 26.6 +/- 7.4 versus 7.6 +/- 2.0 pmol/h, P < 0.02). In 8% O2, control DACB (576 +/- 133 pmol/CB) was significantly higher than AMPT or haloperidol (respectively 228 +/- 29.6 and 246 +/- 49.9 pmol/CB, P < 0.01) and control DAr (234 +/- 72.3 pmol/h) was also significantly higher than AMPT or haloperidol (respectively 28.8 +/- 5.2 and 40.6 +/- 11.4 pmol/h, P < 0.01). Finally, DAr was significantly larger in 8% O2 than in 100% O2 in control and AMPT groups (P < 0.01), but not in the haloperidol group. The increase in DAr by haloperidol in the resting CB is consistent with the blockade of D2's regulating DA release. The decreased DAr in 8% O2 after AMPT suggests that increased DA synthesis contributes to maintain DA secretion by the type I cell exposed to short term hypoxia. The lack of difference in DAr between 8% O2 and 100% O2 after haloperidol probably reflects non specific--i.e., D2 independent--effect of micromolar concentration of haloperidol on DA synthesis and/or sodium-calcium exchangers during hypoxia. PMID- 9224541 TI - Electrophoretic separation of myosin heavy chain isoforms in the human m. vastus lateralis: references to reproducibility and relationships with force, electromechanical delay, fibre conduction velocity, endurance and electromyography. AB - The aim of this investigation was to determine the relationship between muscle performance and the myosin heavy chain (MHC) composition and the reliability of electrophoretically determined MHC compositions. A total of thirty-one male subjects participated in the experiments. Maximal voluntary isometric contractions (MVC) of the knee extensors were performed at an arbitrary knee angle of 90 degrees and the following variables were recorded: maximal isometric force, muscle fibre conduction velocity (MFCV), electromechanical delay (EMD), maximal rate of force development (MRFD), median frequency of EMG (MF) and iEMG. Static isometric contractions of the knee extensors were held at an angle of 90 degrees using contractile forces of 10%, 50% and 100% MVC, respectively. These tests were conducted on separate days. Muscle biopsy samples were obtained from the left m. vastus lateralis before MVC and static endurance tests. MHC protein isoform differences were determined through sodium dodecyl-polyacrylamide gel electrophoresis (SDS-PAGE) followed by densitometric analysis. Type I-MHC compositions of the m. vastus lateralis ranged from 20-68% with a mean of 49 +/- 18%, mean type IIa-MHC and type IIb-MHC percentages were 35 +/- 16% and 16 +/- 10%, respectively. MHC compositions of duplicate biopsy samples were not significantly different from that of original samples. The coefficients of variation calculated for duplicate biopsy samples suggested reasonable reproducibility for MHC isoform differentiation for type I-MHC and type-II MHC composition (CV = 12.6%). Differentiation between type IIa-MHC and type IIb-MHC was not always clear using the densitometric traces. Subjects with higher percentages of type II-MHC displayed significantly faster MFCV (r = 0.67, P < 0.1), isometric force development (r = 0.68, P < 0.1) and shorter periods of EMD (r = -0.72, P < 0.05). There was also a tendency toward faster MRFD in these subjects although results did not reach significance. Endurance times for isometric contractions held at 10%, 50% and 100% MVC to exhaustion were not correlated with MHC composition. No relationships between II-MHC composition and MF or iEMG were observed. It was suggested that surface electromyographic recordings obtained during isometric MVC did not reflect underlying differences in muscle fibre composition. PMID- 9224542 TI - Relationship between acid neutralization capacity of saliva and gastro oesophageal reflux. AB - Saliva is an important factor in neutralization of oesophageal acid exposure, clinically manifested as gastrooesophageal reflux (GOR). The aim of this study is to compare the composition and the "capacity in acid neutralization" (CAN) of saliva in controls and patients suffering of GOR. We compared the composition of saliva from 56 patients who had symptoms of GOR with that of saliva from 20 healthy control subjects. After a standardized 24-hour period of pH-monitoring, 39 patients had normal pH reflux scores (normal acid score: NAc-GOR) and 17 had abnormal pH reflux scores (pathological acid score: PAc-GOR). Then, following a 10-h fast, total saliva was collected during ten minutes in all patients and in the healthy control subjects. The composition of the saliva samples was analysed and the titration curve was determined on 200 microliters aliquots by successive addition of 5 microliters volumes of 0.1 N Hcl. The GOR patients had significantly lower salivary concentrations of Na+ and of both free and bound sialic acids and had higher salivary concentrations of inorganic phosphates than the controls. These disorders were more marked in PAc-GOR patients. Initial pH was 7.43 +/- 0.43 in controls, 7.35 +/- 0.45 in NAc-GOR patients, and 6.91 +/- 0.53 in PAc-GOR patients. In the beginning of the titration curve, PAc-GOR patients were significantly different from NAc-GOR patients and from controls. Saliva of both groups of patients presented significant differences in the acidic portion of the titrations curves, at high volumes of added HCl. These data show that the composition of saliva was modified in patients with GOR disease compared to that of normal subjects. A difference in titration curves was also observed with a higher acidic buffering capacity in these GOR patients. The modifications in saliva composition suggest a role for inorganic phosophates in the acid neutralization capacity observed in GOR, perhaps linked with a adaptation to chronic acid exposure. PMID- 9224543 TI - Plasma myosin and creatine kinase time-course after a concentric-eccentric field exercise. AB - The effect of a concentric (uphill run)-eccentric (downhill run) field exercise of 22.3 km long was examined in five healthy male volunteers to compare the time course of changes in plasma creatine kinase (CK) activity and beta myosin heavy chain (beta MHC) concentration observed during the recovery. CK and beta MHC were examined in blood sampled before exercise, immediately and 5 hours after exercise ceased. Screenings were conducted 1, 2, 3, 4, 5 and 7 days later. For every subject, the peak of plasma CK was transient and observed within the first 24 hours of recovery. In contrast to CK changes, plasma beta-MHC elevation was delayed and the peak, also transient, was observed the second or the third day after the exercise. The highly significant relationship between individual values of CK and beta MHC (P < 0.001) demonstrates that beta MHC could be used as a marker of skeletal muscle damage after acute exercise. PMID- 9224544 TI - Effect of endotoxemia on plasma and tissue levels of nitric oxide metabolites and guanidino compounds. AB - The effect of endotoxemia on the levels of amino acids, nitrates, nitrites and guanidino compounds was investigated. Plasma levels of nitrate and nitrite were significantly increased indicating increased production of nitric oxide during endotoxemia. Plasma concentrations of alanine, glutamine, leucine, methionine, phenylalanine, proline and taurine were also significantly elevated. These results indicate that endotoxin produces a hypercatabolic state. The plasma concentration of arginine was significantly decreased whereas the concentrations of ornithine and urea, the catabolites of arginine were increased. Decreased plasma arginine coupled with increased plasma ornithine and urea indicate that arginine catabolism is increased and arginine synthesis is decreased during endotoxemia. Plasma levels of creatine, creatinine, guanidine and guanidinosuccinic acid were significantly elevated whereas homoarginine levels were significantly decreased. Nitric oxide synthase utilizes arginine as well as homoarginine as substrates. The decreased concentration of both substrates may be related to alterations in nitric oxide synthase activity during endotoxemia. These results suggest that in addition to nitric oxide, other catabolites of arginine such as guanidino compounds may be important in the pathophysiology of endotoxemia. Because of the marked increase in guanidinosuccinic acid, a known uremic toxin, we speculate that guanidinosuccinic acid may be important in the pathophysiology of endotoxemia. PMID- 9224545 TI - Transepithelial potential difference of a single gill filament isolated from the crayfish Astacus leptodactylus Esch.: a new method. AB - A new method is described that allows in vitro perfusion and transepithelial electrical potential measurements of a single filament (3-5 mm long; 200 microns in diameter) isolated from the podobranch of the crayfish Astacus leptodactylus. An electrophysiological study was carried out on the preparation to validate this technique. The good physiological quality of the isolated filament preparation has been established and results of continuous measurements of the potential difference under two perfusion conditions are reported. Filaments were perfused with Van Harreveld physiological saline inside and either with Van Harreveld saline or artificial fresh water outside. Large potential differences up to 150 mV between inside and outside of the filament were recorded. When filaments were symmetrically perfused, the behavior of the electrical potential difference allowed two populations of filaments to be distinguished. PMID- 9224546 TI - Regional variations in electrical and ion transport properties along the isolated intestine of the frog Rana esculenta. AB - Anterior, posterior and colon regions of isolated intestines of the frog Rana esculenta were studied in Ussing chambers under short-circuit conditions. Each region presented a serosa-positive potential which decreased upon longer incubation with no significant change in resistance. The colon displayed higher transepithelial potential (initial mean: 11.4 mV) and resistance (165.cm 2) than the proximal parts (initial mean: ca. 2 mV and 120-80 .cm 2). Bilateral substitution of Na+ by NMDG (N-methyl-D-glutamine) or of Cl- by gluconate induced large and sustained decreases in potential and current, which were reversed in the anterior and posterior intestine and abolished in colon, indicating strict dependence upon the presence of both Na+ and Cl-. The mucosal membranes showed the presence of amiloride-sensitive Na+ sites (with drug efficiency higher in colon). Na+/K+/2Cl- cotransport (current decreased by about 50% by bumetanide in anterior and posterior regions only), Cl- permeability or channels inhibited by diphenylamine-2-carboxylate, DPC (similar decreases as by bumetanide). In either chamber 5 mM BaCl2 induced 20-42% inhibition of current, indicating the occurrence of barium-sensitive K+ channels in both apical and basolateral membranes (more markedly on serosal side) in all three intestinal regions. Finally, current increase by IBMX and theophylline designate the colon as a target for adenylate cyclase stimulating hormones. PMID- 9224547 TI - Action potential configuration in heart papillary muscles from female rats in different thyroid states. AB - We have studied the effects of thyroidectomy and the in vivo administration of different triiodothyronine (T3) doses in thyroidectomized female rats on electrophysiological properties, measured in vitro, of the anterior and posterior papillary muscle fibers from the right ventricle. In each thyroid state, the action potential duration (APD) measured by stimulating at 1 Hz was shorter for the posterior papillary muscle. APD from both preparations was found significantly lengthened in thyroidectomized animals in comparison to euthyroid controls. APD was shortened owing to treatment of thyroidectomized rats with T3 doses up to 10 micrograms/100 g body weight every second day. Treatment with larger doses of T3 tended to restore the values of APD found for ventricular fibres from both controls and thyroidectomized animals treated with substitutive T3 doses (5 micrograms/100 g body weight every second day). As the stimulation rate was increased from 1 to 5 Hz, APD increased in both preparations of all groups. The changes were of different amounts but the APD difference between the rat groups, which were significant at 1 Hz, remained significant at 5 Hz, while the differences between anterior and posterior preparations were cancelled in animals treated with 50 micrograms of T3 and reversed in those treated with 100 micrograms. PMID- 9224548 TI - Effect of activators and inhibitors of K+ channels on insulin secretion in the amphibian pancreas. AB - The aim of this study was to obtain pharmacological evidence for the presence and participation of K+ channels in amphibian pancreatic islets. Pancreases from the toad Bufo arenarum were thus incubated with activators or blockers of K+ channels and the immunoreactive insulin released into the medium was measured by radioimmunoassay. Two K(+)-ATP channel openers (diazoxide and BPDZ44) inhibited; while a K(+)-ATP channel blocker (tolbutamide) and metabolizable sugars (glucose, glyceraldehyde) significantly stimulated the output of insulin. Although a nonmetabolizable sugar (galactose) failed to increase insulin release, dinitrophenol decreased the secretagogue effect of glucose. By contrast, although somatostatin and clonidine blocked the release of insulin, tetraethylammonium significantly stimulated secretion. For each compound tested, the effects on both insulin secretion and B-cell K+ channel activity were similar to those observed in the mammalian pancreas. These findings point to the existence of mammalian like K+ channels in the B-cells of some amphibians. PMID- 9224549 TI - Effects of the amount and type of dietary fat on exocrine pancreatic secretion in dogs after different periods of adaptation. AB - Mongrel dogs were fed, from weaning to 6 months of age, on one of two 9% lipid diets that differed only in the type of fat content (sunflower oil or virgin olive oil) to study their effects on exocrine pancreatic secretion, in the basal period and in response to food. In addition, the results were compared with those obtained in a previous work performed by us on dogs adapted for 8 months to diets containing a higher (15%) amount of the same dietary fats to further evaluate the influence of the amount of dietary fat and the length of the adaptation period. The results from the present study show that both the volume and bicarbonate secreted in the absence of stimuli are unaffected by the quality of dietary fat. In contrast, in response to food, the pancreatic juice flow and the bicarbonate output were significantly higher in the group of animals given the sunflower oil diet. The differences seem to be related with the oleic acid content in the diets and the effectiveness of this fatty acid in triggering the release of inhibitory peptides such as pancreatic polypeptide and peptide YY. The comparison between the results from our present and previous studies supports the afore-mentioned hypothesis and confirms the existence of a clear influence of the amount and type of dietary fat, especially the oleic acid content, upon the pancreatic response to food, without ruling out a role for the duration of the adaptation period. PMID- 9224550 TI - Lipid modifications of microsomes isolated from villus and crypt zones of bovine intestinal mucosa: relationship with fatty acid binding protein. AB - The present studies were conducted to examine the fatty acid composition of microsomal lipids of bovine small intestine. Microsomes and cytosol were isolated from mucosal scrapings enriched with villus and crypts cells, and the following studies were conducted to: 1) analyse fatty acids from microsomal lipids; 2) incorporate 1-14C oleic acid to microsomal lipids; and 3) bind 1-14C oleic acid to cytosolic proteins of villus and crypt zone. The results of these studies demonstrated that: (1) the major unsaturated fatty acids of microsomes were oleic (C18:1 n-9), linoleic (C18:2 n-6) and arachidonic acids (C20:4 n-6), which increased from crypt to villus tip of the bovine intestinal mucosa; (2) gel filtration chromatography indicated that the low-molecular weight cytosolic proteins obtained from superficial mucosal scrapings contained the greatest oleic acid binding activity; (3) the incorporation of 1-14C oleic acid to microsomes was higher in phospholipids, triglycerides and cholesterol esters from villus than in crypts zones; (4) the protein content of cytosol and microsomes was longer in villus zones than in crypts zones; (5) the peroxidizability index showed the highest value in villus microsomes. PMID- 9224551 TI - Dynamics of two-component biochemical systems in interacting cells; synchronization and desynchronization of oscillations and multiple steady states. AB - Systems of interacting cells containing a metabolic pathway with an autocatalytic reaction are investigated. The individual cells are considered to be identical and are described by differential equations proposed for the description of glycolytic oscillations. The coupling is realized by exchange of metabolites across the cell membranes. No constraints are introduced concerning the number of interacting systems, that is, the analysis applies also to populations with a high number of cells. Two versions of the model are considered where either the product or the substrate of the autocatalytic reaction represents the coupling metabolite (Model I and II, respectively). Model I exhibits a unique steady state while model II shows multistationary behaviour where the number of steady states increases strongly with the number of cells. The characteristic polynomials used for a local stability analysis are factorized into polynomials of lower degrees. From the various factors different Hopf bifurcations may result in leading for model I, either to asynchronous oscillations with regular phase shifts or to synchronous oscillations of the cells depending on the strength of the coupling and on the cell density. The multitude of steady states obtained for model II may be grouped into one class of states which are always unstable and another class of states which may undergo bifurcations leading to synchronous oscillations within subgroups of cells. From these bifurcations numerous different oscillatory regimes may emerge. Leaving the near neighbourhood of the boundary of stability, secondary bifurcations of the limit cycles occur in both models. By symmetry breaking the resulting oscillations for the individual cells lose their regular phase shifts. These complex dynamic phenomena are studied in more detail for a low number of interacting cells. The theoretical results are discussed in the light of recent experimental data on the synchronization of oscillations in populations of yeast cells. PMID- 9224552 TI - Information processing and symmetry-breaking in memory evolutive systems. AB - The aim of this paper is to evaluate the role of symmetry and symmetry-breaking processes on the complex information processing developed by hierarchical evolutionary natural systems, such as biological, neural, social or cultural systems. The study is conducted in the frame of the Memory Evolutive Systems, which give a mathematical model of these systems. The dynamics of a MES is modulated by the competition between a net of internal regulation centers which act apart-but encode overlapping strategies which have to be equilibrated. The main characteristics of these systems, at the root of their complexity and adaptability, is a symmetry-breaking in the passage from a higher (or macro) level to a lower (or micro) level: several disparate sub-systems with different comportments at the micro level can be undistinguishable at the higher macro level because of a similar macro behavior (Multiplicity Principle). It is responsible for the development of a dialectics between heterogeneous regulation centers, and for the emergence in time of more and more complex objects. An application to neural systems vindicates an emergentist dynamical reduction of mental states to physical states. PMID- 9224553 TI - Constancy, uniformity and symmetry of living systems: the computational functions of morphological invariance. AB - Living systems, whether organs or entire organisms, display various forms of morphological invariance. Why? These phenomena are studied here from the Proto Cognitive perspective, according to which evolution proceeds by processing information about the environment. The evolution of constancy, uniformity and symmetry is studied in detail. The study later focuses on the invariances of the plant's leaves and it is proposed that these invariances play a crucial role in the plant's development of individual form. Experiments are proposed to test these hypotheses. Morphological invariance is further examined in the light of thermodynamics and information theory. New thermodynamic restrictions are imposed on the processes of measurement and information recording. Uniformity and symmetry are shown to meet these restrictions. PMID- 9224554 TI - Is symmetry informative? AB - Is symmetry informative? The answer is both yes and no. We examine what information and symmetry are and how they are related. Our approach is primarily mathematical, not because mathematics provides the final word, but because it provides an insightful and relatively precise starting point. Information theory treats transformations that messages undergo from source to destination. Symmetries are information that leave some property of interest unchanged. In this respect the studies of information and symmetry can both be regarded as a Quest for the identity transformation. PMID- 9224555 TI - A theoretical integration of Cambrian explosion and post-Permian quiescence. AB - Trophic dynamics is internally causative. Each trophic level is causative in initiating changes in trophic flow either as a supplier towards the upper level or as a consumer towards the lower. Resource presentation followed by its subsequent exploitation makes suppliers causative, while resource exploitation followed by its subsequent presentation makes consumers causative. Trophic dynamics of supplier domination gradually alternates with that of consumer domination while being punctuated by occasional mass extinctions due to depletion of the resources towards the lowest trophic level. The Cambrian explosion could be associated with trophic dynamics of supplier domination, whereas the post Permian quiescence with that of consumer influencing supplier domination. PMID- 9224556 TI - Computer science and biology--the German Conference on Bioinformatics (GCB'96). PMID- 9224557 TI - RNA-RNA interaction and gene splicing. AB - The precise excision of intervening sequences during RNA splicing is an interesting example of the high degree of specificity involved in biosynthesis processes. Self-splicing RNA precursors achieve this specificity primarily through intramolecular interactions whereas all other types of RNA splicing requires interaction between cellular factors and specific recognition signals in the RNA precursor. About twelve years ago, the in vitro splicing system was developed and a general scheme of the pre-mRNA was proposed (Hernandez and Keller, 1983; Krainer et al., 1984; Lin et al., 1985; Padgett et al., 1984; Ruskin et al., 1984). A fundamental question in the splicing field is how the 5' and 3' splice sites are recognized and paired during the splicing reaction. Recent work in the splicing field has established that a network of RNA interactions may form the structural foundation of the spliceosomes. Possible solutions to many unsolved puzzles are getting attention. RNA-RNA interactions now appear to underlie many aspects of substrate recognition, reaction partner juxtaposition and catalysis. In this article we have presented the latest mechanisms involved in the pre-mRNA splicing and their implication in applied research including cancer. PMID- 9224558 TI - Cytotoxicity to tumors by alpha, beta-dihydric long-chain fatty alcohols isolated from esterolysates of uncytotoxic sheep cutaneous wax: the dependence on the molecular hydrophobicity balance of N- or iso-alkyl moiety bulkiness and two hydroxyl groups. AB - Wool fatty alcohols (WF-Alc; C10-C33), separated by esterolysis of wool grease secreted from sheep sebaceous gland, inhibited growth of mouse Ehrlich ascites carcinoma (EAC) cells in contrast to no inhibition by unesterolysed wool grease. WF-Alc was fractionated by molecular distillation and subsequent octadecylsilica (ODS) gel liquid chromatography, showing that most of the growth-inhibitory activity was found in the most hydrophilic fraction with the lowest boiling-point (MW 200-300; C12-C20), ODS-HPLC of the fraction showed that most of the activity resided in two homogeneous fractions identified by GC-MS and 13C-/1H-NMR as alpha, beta-dihydric saturated fatty alcohols such as 1,2-hexadecanediol (n C16(OH)2) and 16-methyl-1,2-heptadecanediol (iso-C18(OH)2), respectively. EAC cells implanted into mice were inhibited markedly by n-C16(OH)2 possessing a cytolysing ability and slightly by iso-C18(OH)2, but hardly by other alkyl-alpha, beta-diols (C12-C24) contained in WF-Alc. Thus, the antitumor activity of WF-Alc was exhibited only after saponification of uncytotoxic wool grease, showing necessity of unesterified hydroxyl groups, and was dependent upon the molecular hydrophobicity balance attributed to both hydroxyl groups and n- or iso-alkyl moiety bulkiness specified out of diverse species of fatty alcohols contained in WF-Alc. PMID- 9224560 TI - Reverse-micelle model: pH, electromagnetic field and inhibitor enzyme interaction. AB - The reverse micelle is one of many models thought to have properties more nearly resembling the biological cellular environment, than does the traditional dilute solution biochemical reaction system. In order to evaluate the results of EMF perturbation of enzyme-catalyzed reactions, the description of the AOT reverse micelle model, with respect to its internal pH, effect of chemical inhibitors, temperature, and electromagnetic-field perturbation has herein been extended. Acetylcholinesterase and NADPH cytochrome-P450 reductase, reacting within the AOT reverse-micelle, exhibit a temperature vs. activity profile equivalent to the same reaction in a buffered dilute-solution environment. In reverse micelles, some inhibitors of AChE (propidium, and d-tubocurarine) have much less effect upon indophenol-acetate hydrolysis than they do in a dilute solution environment. Other inhibitors act in the same manner within the structured environment of the reverse micelle as in the conventional dilute solution reaction model. These differences are explicable in terms of mechanism of action of the individual inhibitors. Perturbation by low-intensity microwave fields has a similar inhibitory effect upon dilute-solution reactions, as those in the 'low-water activity' environment of the reverse micelle. However, the interactions between physical and chemical perturbants are differently limited by the structure of the aqueous phase of the reverse micelle. pH of the 'internal' reverse-micelle environment is a function of the availability of H-ions supplied by system components. Use of indicator dyes show that the low-molarity buffers which are compatible with reverse-micelle stability, are often insufficient to maintain a constant pH. Too, in the reverse micelle, reaction rate, for proton yielding reactions, is dramatically greater than the rate of the same reaction in dilute solution at the same acidic pH. PMID- 9224559 TI - Preneoplasia-associated expression of calcyclin and of binding sites for synthetic blood group A/H trisaccharide--exposing neoglycoconjugates in human lung. AB - Development of preneoplastic lesions in human lung is supposed to be accompanied with alterations of distinct biochemical features which might functionally be crucial for this alteration. To contribute to the definition of such determinants in peripheral lung parenchyma, the files of the Department of Pathology, Thoraxklinik, (a total of 2890 cases) were screened for respective tissue specimens. Seventy one cases with complete clinical documentation were found and an age-, sex-, and disease-matched control group was formed. When compared to control group patients, especially the tumor free cases with preneoplastic aberrations revealed a history of exposure to external noxes. Several probes with assumed relevance were tested with the panel of specimens for both groups, focussing on comparative analysis of alveolar lining cells. In addition to labelled neoglycoconjugates which include tissue lectin-seeking probes that expose mono-, di- and blood group-related trisaccharides, presence of calcium- and annexin-binding calcyclin, of complement component C5b, of the lymphokine macrophage migration inhibitory factor, and of ligands of the serum amyloid P component was evaluated. Compared to normal cells at the alveolar surface in controls, the preneoplastic cells displayed an apparent down-regulation of expression of A/H-trisaccharide-specific binding sites and an upregulation of expression of calcyclin. These three characteristics correlated with the phenotypic alterations and encourage further studies to elucidate the functional significance of reduced expression of the glycoligand-specific sites and the presence of this member of the S100-family of Ca(2+)-binding proteins. PMID- 9224561 TI - Inhibition of the proliferation of Ehrlich ascites tumor cells by hydrostatic pressure. AB - The effect of high pressure on the viability of Ehrlich ascites tumor cells was examined. The tumor cells were subjected to various pressures (0.1-150 MPa) for 30 min at 37 degrees C. The viability of pressure-treated cells was examined by the dye exclusion method. The number of stained cells increased significantly at pressures above 130 MPa. In addition, the pressure-treated cells were intraperitoneally inoculated into the mice. The tumor cells which were subjected to pressures below 110 MPa proliferated in the peritoneal cavity of the mice, so that the mice died. In contrast, the mice, which were inoculated with the tumor cells treated at pressures above 130 MPa, remained alive. These results suggest that the destruction of the tumor cells begins to occur at about 130 MPa. PMID- 9224562 TI - Detection of breast tumor associated mucin epitope on CAMA cell line using monoclonal antibody G3F1 generated against HMFG membrane. AB - The expression of breast tumor associated mucin epitope on CAMA cell line was detected employing the mouse MAB G3F1 generated against HMFG membrane. Immunocytochemical studies revealed that MAB G3F1 strongly reacted with 85% of breast cancer tissue sections with specific staining of apical cell membrane of malignant epithelial cells. The mucin antigen recognised by MAB G3F1 was detected by selectively extracting high molecular weight glycoprotein antigen from HMFG membrane using lectin affinity chromatography and gel filtration chromatography. Immunoprecipitation studies revealed that MAB G3F1 recognized a high molecular weight glycoprotein with an approximate molecular weight of 300kd. The expression of MAB G3F1 reactive antigen on CAMA cells was detected by immunocytochemistry and by immumoprecipitating 300kd antigen from 125I labelled Tx100 solubilized extract of CAMA cells. The results from these investigations suggest that CAMA cells express MAB G3F1 reactive antigen with tumor associated epitope, similar to tumor associated mucin epitope of HMFG membrane. PMID- 9224563 TI - Invasive properties of cadmium-resistant human fibrosarcoma HT-1080 cells. AB - Invasive properties of tumor cells having acquired heavy metal resistance were investigated. We selected the cadmium-resistant (Cd-R) cells from human fibrosarcoma HT-1080 cells. Total metallothionein levels in cytosol of HT-1080 Cd R cells were significantly higher than original lines, and were of a highly resistant potency to cytotoxicity of cisplatin, as well as heavy metals. The HT 1080 Cd-R cells showed higher invasiveness into recombinant basement membrane Matrigel. However, HT-1080 Cd-R cells were inferior in locomotion ability. Significant differences in adhesive ability to extracellular matrix proteins were not observed between HT-1080 and HT-1080 Cd-R cells. High invasiveness of HT-1080 Cd-R cells was caused by their extremely strong enzymatic activities. High level of 92kDa matrix metalloproteinase-9 (MMP-9) was recognized from the conditioned medium of HT-1080 Cd-R cells, whereas 72kDa MMP-2 was secreted equally from both cell lines. Our investigation suggests that drug resistance acquired through the mechanisms of cellular metal-tolerance may promote malignancy and tumor metastasis during cancer chemotherapy. PMID- 9224564 TI - Mitochondrial and cytosolic rhodanese from liver of DAB-treated mice. III. Inhibition kinetic studies. AB - Rhodanese (thiosulphate:cyanide sulphurtransferase) shows distinctive mitochondrial and cytoplasmic activities in several models of tumorigenesis. To investigate the basis for these differences, the enzyme was purified from mitochondrial and cytosolic liver fractions of mice treated with the carcinogen p dimethyl-aminoazobenzene (DAB) and some inhibition kinetic studies were carried out. When both substrates were assayed at inhibitory levels, non-competitive inhibition was observed for the second substrate at variable concentrations, the reversible connection between both substrates was attained by the instability of the second enzyme form. It is suggested that the enzyme might be changing from an unstable ES form to a more stable sulphur substituted intermediate as a consequence of DAB treatment. Sulphite was a competitive inhibitor vs thiosulphate for rhodanese isolated from normal liver and a hyperbolic activator for the enzyme isolated from liver of DAB-treated animals. PMID- 9224565 TI - The structural basis for the specificity of pyridinylimidazole inhibitors of p38 MAP kinase. AB - BACKGROUND: The p38 mitogen-activated protein (MAP) kinase regulates signal transduction in response to environmental stress. Pyridinylimidazole compounds are specific inhibitors of p38 MAP kinase that block the production of the cytokines interleukin-1beta and tumor necrosis factor alpha, and they are effective in animal models of arthritis, bone resorption and endotoxin shock. These compounds have been useful probes for studying the physiological functions of the p38-mediated MAP kinase pathway. RESULTS: We report the crystal structure of a novel pyridinylimidazole compound complexed with p38 MAP kinase, and we demonstrate that this compound binds to the same site on the kinase as does ATP. Mutagenesis showed that a single residue difference between p38 MAP kinase and other MAP kinases is sufficient to confer selectivity among pyridinylimidazole compounds. CONCLUSIONS: Our results reveal how pyridinylimidazole compounds are potent and selective inhibitors of p38 MAP kinase but not other MAP kinases. It should now be possible to design other specific inhibitors of activated p38 MAP kinase using the structure of the nonphosphorylated enzyme. PMID- 9224566 TI - Iterative type II polyketide synthases, cyclases and ketoreductases exhibit context-dependent behavior in the biosynthesis of linear and angular decapolyketides. AB - BACKGROUND: Iterative type II polyketide synthases (PKSs) produce polyketide chains of variable but defined length from a specific starter unit and a number of extender units. They also specify the initial regiospecific folding and cyclization pattern of nascent polyketides either through the action of a cyclase (CYC) subunit or through the combined action of site-specific ketoreductase (KR) and CYC subunits. Additional CYCs and other modifications may be necessary to produce linear aromatic polyketides. The principles of the assembly of the linear aromatic polyketides, several of which are medically important, are well understood, but it is not clear whether the assembly of the angular aromatic (angucyclic) polyketides follows the same rules. RESULTS: We performed an in vivo evaluation of the subunits of the PKS responsible for the production of the angucyclic polyketide jadomycin (jad), in comparison with their counterparts from the daunorubicin (dps) and tetracenomycin (tcm) PKSs which produce linear aromatic polyketides. No matter which minimal PKS was used to produce the initial polyketide chain, the JadD and DpsF CYCs produced the same two polyketides, in the same ratio; neither product was angularly fused. The set of jadABCED PKS plus putative jadl CYC genes behaved similarly. Furthermore, no angular polyketides were isolated when the entire set of jad PKS enzymes and Jadl or the jad minimal PKS, Jadl and the TcmN CYC were present. The DpsE KR was able to reduce decaketides but not octaketides; in contrast, the KRs from the jad PKS (JadE) or the actinorhodin PKS (ActIII) could reduce octaketide chains, giving three distinct products. CONCLUSIONS: It appears that the biosynthesis of angucyclic polyketides cannot be simply accomplished by expressing the known PKS subunits from artificial gene cassettes under the control of a non-native promoter. The characteristic structure of the angucycline ring system may arise from a kinked precursor during later cyclization reactions involving additional, but so far unknown, components of the extended decaketide PKS. Our results also suggest that some KRs have a minimal chain length requirement and that CYC enzymes may act aberrantly as first-ring aromatases that are unable to perform all of the sequential cyclization steps. Both of these characteristics may limit the widespread application of CYC or KR enzymes in the synthesis of novel polyketides. PMID- 9224568 TI - Rational design of allosteric ribozymes. AB - BACKGROUND: Efficient operation of cellular processes relies on the strict control that each cell exerts over its metabolic pathways. Some protein enzymes are subject to allosteric regulation, in which binding sites located apart from the enzyme's active site can specifically recognize effector molecules and alter the catalytic rate of the enzyme via conformational changes. Although RNA also performs chemical reactions, no ribozymes are known to operate as true allosteric enzymes in biological systems. It has recently been established that small molecule receptors can readily be made of RNA, as demonstrated by the in vitro selection of various RNA aptamers that can specifically bind corresponding ligand molecules. We set out to examine whether the catalytic activity of an existing ribozyme could be brought under the control of an effector molecule by designing conjoined aptamer-ribozyme complexes. RESULTS: By joining an ATP-binding RNA to a self-cleaving ribozyme, we have created the first example of an allosteric ribozyme that has a catalytic rate that can be controlled by ATP. A 180-fold reduction in rate is observed upon addition of either adenosine or ATP, but no inhibition is detected in the presence of dATP or other nucleoside triphosphates. Mutations in the aptamer domain that are expected to eliminate ATP binding or that increase the distance between aptamer and ribozyme domains result in a loss of ATP-specific allosteric control. Using a similar design approach, allosteric hammerhead ribozymes that are activated in the presence of ATP were created and another ribozyme that can be controlled by theophylline was created. CONCLUSIONS: The catalytic features of these conjoined aptamer-ribozyme constructs demonstrate that catalytic RNAs can also be subject to allosteric regulation-a key feature of certain protein enzymes. Moreover, by using simple rational design strategies, it is now possible to engineer new catalytic polynucleotides which have rates that can be tightly and specifically controlled by small effector molecules. PMID- 9224567 TI - Biosynthesis of vitamin B12: the multi-enzyme synthesis of precorrin-4 and factor IV. AB - BACKGROUND: In order to study the biosynthesis of vitamin B12, it is necessary to produce various intermediates along the biosynthetic pathway by enzymic methods. Recently, information on the organisation of the biosynthetic pathway has permitted the selection of the set of enzymes needed to biosynthesise any specific identified intermediate. The aim of the present work was to use recombinant enzymes in reconstituted multi-enzyme systems to biosynthesise particular intermediates. RESULTS: The products of the cobG and cobJ genes from Pseudomonas denitrificans were expressed heterologously in Escherichia coli to afford good levels of activity of the corresponding enzymes, CobG and CobJ. Aerobic incubation of precorrin-3A with the CobG enzyme alone yielded precorrin 3B. When CobJ and S-adenosyl-L-methionine were included in the incubation, the product was precorrin-4. Both precorrin 3B and precorrin-4 are known precursors of vitamin B12 and their availability has allowed new mechanistic studies of enzymic transformations. CONCLUSIONS: Our results show that the expression of the CobG and CobJ enzymes has been successful, thus facilitating the biosynthesis of two precursors of vitamin B12. This lays the foundation for the structure determination of CobG and CobJ as well as future enzymic experiments focusing on later steps of vitamin B12 biosynthesis. PMID- 9224569 TI - RNA as a drug target. AB - Historically, the pharmaceutical industry has focused on proteins, rather than nucleic acids, as drug targets. But recent advances in the fields of RNA synthesis, structure determination and therapeutic target identification make the systematic exploitation of RNA as a drug target a realistic goal. PMID- 9224570 TI - Methionine aminopeptidase (type 2) is the common target for angiogenesis inhibitors AGM-1470 and ovalicin. AB - BACKGROUND: Angiogenesis, the formation of new blood vessels, is essential for tumor growth. The inhibition of angiogenesis is therefore emerging as a promising therapy for cancer. Two natural products, fumagillin and ovalicin, were discovered to be potent inhibitors of angiogenesis due to their inhibition of endothelial cell proliferation. An analog of fumagillin, AGM-1470, is currently undergoing clinical trials for the treatment of a variety of cancers. The underlying molecular mechanism of the inhibition of angiogenesis by these natural drugs has remained unknown. RESULTS: Both AGM-1470 and ovalicin bind to a common bifunctional protein, identified by mass spectrometry as the type 2 methionine aminopeptidase (MetAP2). This protein also acts as an inhibitor of eukaryotic initiation factor 2alpha (elF-2alpha) phosphorylation. Both drugs potently inhibit the methionine aminopeptidase activity of MetAP2 without affecting its ability to block elF-2alpha phosphorylation. There are two types of methionine aminopeptidase found in eukaryotes, but only the type 2 enzyme is inhibited by the drugs. A series of analogs of fumagillin and ovalicin were synthesized and their potency for inhibition of endothelial cell proliferation and inhibition of methionine aminopeptidase activity was determined. A significant correlation was found between the two activities. CONCLUSIONS: The protein MetAP2 is a common molecular target for both AGM-1470 and ovalicin. This finding suggests that MetAP2 may play a critical role in the proliferation of endothelial cells and may serve as a promising target for the development of new anti-angiogenic drugs. PMID- 9224571 TI - Mapping out fat profits. PMID- 9224573 TI - Credentialing: the hot potato is in our hands. PMID- 9224572 TI - Engineered cell surfaces: fertile ground for molecular landscaping. AB - The cell surface contains a wealth of information that determines how cells interact with their environment. Methods for directing the cell surface expression of novel protein-based and oligosaccharide-based epitopes are stimulating new directions in biotechnology and biomedical research. PMID- 9224574 TI - How new technology affects practice and patient safety. PMID- 9224575 TI - Experience with laparoscopic leiomyoma coagulation and concomitant operative hysteroscopy. AB - STUDY OBJECTIVES: To evaluate the experiences of women who underwent laparoscopic leiomyoma coagulation (myolysis) alone and those who had myolysis in conjunction with transcervical endomyometrial resection (TEMR), transcervical electrosurgical resection of submucous leiomyomas (TSR), or both. DESIGN: Continuing, prospective observational study with mean (+/- SEM) follow-up of 36.0 +/- 1.2 months (range 18-54 mo). SETTING: Gynecology department of community and teaching hospitals. PATIENTS: One hundred sixty-seven women with symptomatic leiomyomata. INTERVENTIONS: Women complaining of pressure, pain, or both underwent only myolysis. Those with the additional symptom of chronic menorrhagia underwent TEMR, TSR, or both. Nineteen (11.4%) of the 167 women had elective second-look laparoscopy 6.0 +/- 0.3 months (range 6-8 mo) later to evaluate possible adhesion formation. MEASUREMENTS AND MAIN RESULTS: Main outcome measures were control of symptoms, numbers and types of concomitant and subsequent procedures, changes in uterine and leiomyomata volumes, and number of successful pregnancies. Mean total uterine volume of the 167 women decreased from 620 +/- 28.4 cm3 before leuprolide treatment to 131 +/- 7.2 cm3 by 7 to 12 months postoperatively (p <0.0001). Five (3.6%) women had hysterectomies for persistent or recurrent menorrhagia, pain, pressure, or a combination of symptoms (p = 0.01). Pathologic evaluation revealed adenomyosis, leiomyomata, or both. Of 52 women with chronic menorrhagia, 33 (63.5%) developed amenorrhea and 17 (32.7%) developed hypomenorrhea or eumenorrhea; 2 (3.8%) required repeat TEMR. The two women who desired to retain fertility had uncomplicated full-term pregnancies and uneventful vaginal deliveries. CONCLUSIONS: Myolysis alone or in conjunction with TEMR, TSR, or both obviated the need for major surgery in 162 (97.0%) women. Until further studies are concluded, myolysis should be performed selectively in women contemplating pregnancy. PMID- 9224576 TI - Health and fertility outcomes among women surgically treated for endometriosis. AB - STUDY OBJECTIVE: To assess health and fertility status among women after surgical treatment of endometriosis. DESIGN: Prospective study. SETTING: Community-based gynecologic specialty practice. PATIENTS: Two hundred ninety women with newly diagnosed endometriosis. MEASUREMENTS AND MAIN RESULTS: Medical records of all women were abstracted at baseline; self-administered questionnaires were used to collect follow-up data. Most women (68-79%) reported some or great improvement in symptomology after surgical treatment. One hundred twenty-four (53%) of 232 women reported one or more pregnancies, two-thirds of which resulted in live births. Secondary sex ratios were below 1 (range 0.92-0.50), reflecting a female excess. Logistic regression analysis identified previous live birth as the only significant predictor of pregnancy after surgery; advancing maternal age significantly decreased the likelihood of a live birth. CONCLUSIONS: Overall, these women reported improvement in symptoms at follow-up. Operative and clinical findings were not significant predictors of pregnancy likelihood. Prior live birth conferred more than a twofold increase in pregnancy likelihood, whereas advancing age decreased the likelihood. Reasons for reversed sex ratios are unknown but warrant further study. PMID- 9224577 TI - Preliminary prospective observations on the laparoscopic management of endometrial carcinoma using the two-stage approach to aortic lymphadenectomy. AB - STUDY OBJECTIVE: To determine the value of a two-stage approach to laparoscopic aortic lymphadenectomy (ALN) in women with endometrial cancer. DESIGN: Prospective case series. PATIENTS: Twenty-three consecutive, unselected women with endometrial cancer were managed prospectively according to a previously defined protocol. All had laparoscopic hysterectomy, ten required pelvic and one had an aortic lymphadenectomy (ALN). Pelvic lymph node metastases (PLNM) were present in two (20%) and aortic lymph node metastases in one (10%) patient. Mean age was 60; three women were over 80 years old, and two were 78 years old. Mean weight and body mass index were 192 and 33.5, respectively; two women weighed over 300 pounds and another two weighed over 250 pounds. Mean anesthetic time was 3.2 hours, mean blood loss 469 ccs, and mean drop in hemoglobin 2.5 g/dl. One patient was transfused. Median hospital stay was 2 days. One patient had a questionable ileus post-operatively, and another was hospitalized for 10 days to control her diabetes and blood pressure. CONCLUSIONS: By predicating ALN on the presence of PLNM in endometrial cancer, the number of ALN can be reduced without reducing the number of aortic lymph node metastases detected, and laparoscopic management can be extended to morbidly obese women. PMID- 9224578 TI - Transvaginal ultrasonography and hysteroscopy in the diagnosis of endometrial abnormalities. AB - STUDY OBJECTIVE: To investigate the value of transvaginal ultrasonography, aspiration biopsy, and hysteroscopy combined with curettage or directed biopsy in detecting endometrial pathology in women with abnormal uterine bleeding. DESIGN: Prospective, nonrandomized study. SETTING: A university-affiliated hospital. PATIENTS: One hundred twenty-two premenopausal and 78 postmenopausal women with abnormal uterine bleeding. INTERVENTIONS: The women underwent transvaginal ultrasonography (TVS) combined with aspiration Pipelle biopsy. They were scheduled for hysteroscopy and endometrial sampling by curettage or directed biopsy within 4 weeks. MEASUREMENTS AND MAIN RESULTS: Ultrasonographic findings were evaluated on the basis of final diagnoses established by hysteroscopy and histologic examination. The endometrium was measured at its thickest part in the longitudinal plane. In premenopausal women, endometrial thickness was measured during the early proliferative phase of the cycle. Ultrasound examination was considered negative if single-layer thickness was less than 5 mm in the absence of endometrial projections. In all other cases it was classified as positive. For postmenopausal women the cutoff point was 4 mm (single layer). In postmenopausal women with endometrial thickness less than 4 mm, as well as in premenopausal patients with negative TVS, the combination of TVS and aspiration biopsy missed only one case of atypical hyperplasia. In premenopausal patients TVS clearly detected 73% of polyps and myomata, permitting diagnostic and surgical hysteroscopy to be performed at the same time. In postmenopausal women with endometrial thickness 4 mm or greater, aspiration biopsy failed to detect two cases of atypical hyperplasia and one of focal adenocarcinoma. Pipelle sampling was technically infeasible in a woman with endometrial cancer because of a stenotic cervix. It also missed the majority of benign lesions (polyps and myomas). CONCLUSIONS: Transvaginal ultrasound seems to be an excellent initial diagnostic method, with high sensitivity in diagnosing endometrial abnormalities. Its combination with aspiration biopsy seems to be safe in women with a thin endometrium. Hysteroscopy is necessary in postmenopausal women with an endometrium of 4 mm or more, as well as in premenopausal patients with endometrial thickness more than 5 mm (preovulatory phase of the cycle) and in those with suspected polyps or myomas. PMID- 9224580 TI - Comparison of 49 laparoscopic myomectomies with 49 open myomectomies. AB - STUDY OBJECTIVES: To compare the results of open myomectomy with those of laparoscopic myomectomy, and to assess complications, surgical results, total hospital cost, and morbidity associated with each procedure. DESIGN: Retrospective chart review. SETTING: Private practice of one surgeon, and Department of Obstetrics and Gynecology, Rush Medical College, Chicago, Illinois. PATIENTS: Ninety-eight women with symptomatic uterine leiomyomata. INTERVENTIONS: Forty-nine consecutive laparoscopic myomectomies were performed between 1993 and 1995, and 49 open myomectomies were performed between 1983 and 1995. MEASUREMENTS AND MAIN RESULTS: Indications for both procedures were similar, including menometrorrhagia, pelvic pain, and enlarging myomata. Mean operating time for open myomectomies was 133 minutes versus 264 minutes for laparoscopies (p <0.0001). Mean blood loss was 340 ml and 110 ml, respectively (p <0. 001). The greatest blood loss was 1000 ml in the open group and 800 ml in the laparoscopic group. Uterine size at surgery was 12 to 14 weeks in 42.9% of the open group and 9 to 11 weeks in 51% of the laparoscopy group. The open group incurred a total of 272 hospital days versus 29 days in the laparoscopic group (maximum 25 and 3 days, respectively; mean 5.6 and 0.6 days, respectively; p <0.001). The frequency of postoperative complications was higher in the open group (17) than in the laparoscopic group (5, p = 0.0068). Of patients in whom postoperative adhesions were evaluated, the overall frequency of adhesions was lower in the laparoscopic group. Three women in the open group required postoperative transfusions, compared with none in the laparoscopic group. Seven pregnancies have thus far occurred in the laparoscopic group. Three women delivered at term by elective cesarean section, at which no evidence of uterine dehiscence was found. Estimated average cost of each procedure, expressed in April 1995 dollars using the Consumer Price Index, were $14,461 for open myomectomies and $13,814 for laparoscopies (p = 0.65). Linear regression with residual analysis was performed on costs for both groups and revealed significantly increasing time trend for open myomectomies. During the years of this study, the open procedures increased in price at a rate of $868/year. The cost of laparoscopic myomectomies showed no time trend. CONCLUSIONS: Compared with open myomectomy, laparoscopic myomectomy had lower morbidity, no identifiable trend of increasing hospital cost, minimal hospital stay, and fewer complications. PMID- 9224579 TI - The value of hysteroscopic exploration for abnormal uterine bleeding. AB - STUDY OBJECTIVE: To determine sensitivity, specificity, positive predictive value, negative predictive value, and global diagnostic precision of hysteroscopic exploration in the diagnosis of endometrial hyperplasia and adenocarcinoma in women with abnormal uterine bleeding. DESIGN: Retrospective analysis. SETTING: University-affiliated hospital. PATIENTS: One thousand three hundred ninety-eight patients with abnormal uterine bleeding, 57.3% premenopausal and 42.6% postmenopausal. INTERVENTIONS: Diagnostic hysteroscopy and subsequent dilatation and curettage. MEASUREMENTS AND MAIN RESULTS: Endometrium was classified hysteroscopically as normal, atrophic, endometrial hyperplasia, and endometrial carcinoma. Histopathologic diagnosis was performed to determine the efficacy of hysteroscopy in diagnosing endometrial hyperplasia and adenocarcinoma. For endometrial hyperplasia in premenopausal women, sensitivity was 71.8%, specificity 96.4%, and global diagnostic precision 92.5%; in postmenopausal women, respective figures were 85. 1%, 100%, and 97.3%. For diagnosing adenocarcinoma in premenopausal patients, hysteroscopy was 100% sensitive, with specificity 99.4% and global diagnostic precision 99.5%; in postmenopausal women, respective figures were 100%, 99.4%, and 99.5%. CONCLUSIONS: In women with abnormal uterine bleeding, diagnostic hysteroscopy is a basic tool that allows precise diagnosis of endouterine lesions such as polyps and submucous myomas. It also is highly accurate for evaluating endometrial adenocarcinoma and hyperplasia. PMID- 9224581 TI - Vaginoscopic approach to outpatient hysteroscopy. AB - STUDY OBJECTIVE: To evaluate a new method of outpatient hysteroscopy. DESIGN: Prospective observational study. SETTING: Departments of Obstetrics and Gynecology, Medical School of Ioannina, Greece, and Brooklyn Hospital Center, Brooklyn, New York. PATIENTS: Three hundred twenty-four women, 316 of whom were symptomatic and 8 asymptomatic. INTERVENTIONS: Hysteroscopic vaginoscopy with directed endometrial biopsies. MEASUREMENTS AND MAIN RESULTS: The procedure was successful and well tolerated in 211 (65%) women. In 90 (27.9%) women, the procedure was successful but with significant pelvic pain, and in the remaining 23 (7.1%) it was complicated by either vagal reaction or complete intolerance. Hysteroscopic findings were in agreement with histopathologic results in 290 (89.5%) patients. In all 12 women with endometrial adenocarcinoma, hysteroscopic findings were identical with histopathologic results. CONCLUSION: The vaginoscopic approach is effective for outpatient hysteroscopy. PMID- 9224582 TI - Sterilization by open laparoscopy in a private office. AB - Between December 4, 1979, and September 1, 1996, I performed 813 laparoscopic sterilizations in my private office setting. To increase safety and reduce costs, open laparoscopy was done under local anesthesia in every case. The procedure was completed in 811 women. There were no major complications. Minor complications consisted of three superficial wound infections and two failed laparoscopies. PMID- 9224583 TI - Multicenter feasibility study of a new coaxial falloposcopy system. AB - We compared falloposcopy employing a new coaxial system with traditional laparoscopic chromotubation and hysterosalpingography in a prospective, multicenter clinical trial at five tertiary infertility centers. Based on findings at hysterosalpingography or laparoscopic chromotubation, the 16 women (22 tubes) in group 1 had a presumed diagnosis of proximal tubal obstruction, and the 4 (7 tubes) in group 2 had unexplained infertility. Cannulation was successfully achieved in 83.3% of tubes. In group 1, 85% (17/20) of visualized tubes were patent and 35% (7/20) were normal. In group 2, 40% (2/5) of visualized tubes were abnormal. Management was changed in 52.4% of women as a result of falloposcopic findings. Falloposcopy with this new coaxial system allows improved visualization with less bulky and less traumatic instruments. The system provides valuable information regarding the fallopian tube lumen that correlates poorly with that obtained with more traditional techniques. PMID- 9224584 TI - Subtotal vaginal hysterectomy: a new role for an old procedure. AB - Patients with persistent uterine bleeding that is unresponsive to conservative therapy may opt for endometrial ablation over total hysterectomy because of concerns over subsequent sexual dysfunction or other nonclinical issues. Twelve such women with healthy cervices who failed endometrial ablation, and eight candidates for ablation were offered subtotal vaginal hysterectomy as a definitive primary surgical intervention instead of endometrial ablation. Our experience suggests the safety and utility of subtotal vaginal hysterectomy in properly selected patients. Randomized, comparative studies of this technique as an alternative to hysteroscopic ablation or resection may be warranted. PMID- 9224585 TI - A new technique of primary trocar insertion for laparoscopy. AB - A new method of primary trocar insertion exploits the anatomy of the anterior abdominal wall at the umbilicus. The point of fusion between the skin, fascia, and peritoneum is identified, and a tiny incision is made precisely over this point, enabling a small clamp to be introduced directly into the peritoneal cavity. After stretching the opening with this clamp, a 5-mm trocar is introduced into the peritoneal cavity over a blunt probe, and the abdomen is insufflated. The opening is stretched further with a Kelly clamp, and a 10-mm trocar is introduced over a blunt probe. The technique was used in 54 consecutive patients, 20 of whom had prior low vertical incisions. Ten women had very dense periumbilical adhesions, placing at least four at extremely high risk of bowel injury from blind entry. There were no injuries, and the technique is so quick and effective that it is now the author's routine method of trocar insertion for laparoscopy. PMID- 9224586 TI - Laparoscopic resection of a noncommunicating rudimentary uterine horn. AB - Two women had infertility due to a symptomatic unicornuate uterus associated with rudimentary contralateral horn. Both carried successful pregnancies after laparoscopic resection of the horns. PMID- 9224587 TI - Oophoropexy to prevent sequential or recurrent torsion. AB - Laparoscopic oophoropexy may prevent recurrent (repeat torsion of the same ovary) or sequential (subsequent torsion of the contralateral ovary) ovarian torsion. Two adolescent girls with sequential ovarian torsion underwent laparoscopic plication of utero-ovarian ligaments. Neither patient has had recurrence in the 6. 5 and 2 years, respectively, since surgery. Sequential ovarian torsion has been described,1-8 and in almost every instance the authors raised the question of whether or not oophoropexy should have been done at the time of the initial episode of torsion. In virtually every instance the second ovary was removed and the patient rendered menopausal. In two patients with sequential ovarian torsion the ovary was saved and oophoropexy performed laparoscopically in an effort to prevent recurrence. PMID- 9224588 TI - Laparoscopic removal of virilizing hilar cell tumor in a postmenopausal patient. AB - A postmenopausal woman experienced rapidly progressing hirsutism and signs of virilization. Hormone evaluations showed markedly elevated serum testosterone levels and no evidence of excess cortisol or dehydroepiandrosterone sulfate production. A computerized tomographic scan of the adrenals and ovaries was normal, and transvaginal ultrasound revealed a left ovary with a maximum diameter of 3.2 cm. At outpatient laparoscopic bilateral oophorectomy, the left ovary had a benign, 2.5-cm Leydig cell tumor, hilar cell variant. Laparoscopy may be useful in the diagnosis and treatment of select cases of virilizing tumors of the ovary. PMID- 9224589 TI - Ureteral injury after laparoscopic surgery. AB - Ureteral injuries are uncommon but serious complications of laparoscopic pelvic surgery. When unrecognized, patients experience fever, abdominal pain, signs of peritonitis, and leukocytosis usually 48 to 72 hours after the surgical procedure. A 48-year-old woman underwent laparoscopic-assisted vaginal hysterectomy, bilateral salpingo-oophorectomy, and anterior and posterior colporrhapy due to a large, symptomatic uterine myoma. Postoperatively, she suffered from progressive left lower quadrant pain, with drainage of yellowish fluid from the subumbilical puncture wound 5 days after the operation. Significant urinary ascites was present. Intravenous pyelogram revealed injury to the lower third of the left ureter about 3 cm away from the ureterovesical junction. Left-sided percutaneous nephrostomy was performed after transurethral placement of a ureteral stent failed. Reanastomosis of the ureter was performed successfully 3 months later, and the patient fully recovered without compromise of the genitourinary tract. PMID- 9224590 TI - Not so benign endometrial hyperplasia: endometrial cancer after endometrial ablation. AB - The masking or development of endometrial cancer after endometrial ablation is a concern often alluded to in discussions of complications of endometrial ablation. It is necessary to look for a common factor when this complication occurs. Six cases published in peer-reviewed literature were collected to establish a link between the development of endometrial cancer and endometrial ablation. Preexisting endometrial hyperplasia seems to be the common denominator, and should be considered a contraindication to endometrial ablation until more data are collected. PMID- 9224591 TI - CME approved article. Ectopic pregnancy. PMID- 9224593 TI - Representation of cloned genomic sequences in two sequencing vectors: correlation of DNA sequence and subclone distribution. AB - Representation of subcloned Caenorhabditis elegans and human DNA sequences in both M13 and pUC sequencing vectors was determined in the context of large scale genomic sequencing. In many cases, regions of subclone under-representation correlated with the occurrence of repeat sequences, and in some cases the under representation was orientation specific. Factors which affected subclone representation included the nature and complexity of the repeat sequence, as well as the length of the repeat region. In some but not all cases, notable differences between the M13 and pUC subclone distributions existed. However, in all regions lacking one type of subclone (either M13 or pUC), an alternate subclone was identified in at least one orientation. This suggests that complementary use of M13 and pUC subclones would provide the most comprehensive subclone coverage of a given genomic sequence. PMID- 9224592 TI - The world according to Maf. AB - Maf family proteins are so named because of their structural similarity to the founding member, the oncoprotein v-Maf. The small Maf proteins (MafF, MafG and MafK), as do all family members, include a characteristic basic region linked to a leucine zipper (b-Zip) domain which mediate DNA binding and subunit dimerization respectively. The small Maf proteins form homodimers or heterodimers with other b-Zip proteins present in the cell and bind to Maf recognition elements (MARE) in DNA. Since they lack known transcriptional activation domains, the small Maf proteins function either as obligatory heterodimeric partner molecules with numerous large subunits, discussed below, or alternatively as homo or heterodimeric transcriptional repressors. The three small Maf proteins are expressed in a number of overlapping tissues, but their expression profiles nonetheless appear to be under meticulous tissue- and developmental stage specific control. The MARE bears a striking resemblance to the NF-E2 binding sequence. NF-E2 binding sites in the human beta-globin locus control region have been directly implicated as integral components in the circuitry required for eliciting changes in chromatin structure that precede globin gene activation. While the NF-E2 DNA sequence has been shown to be important for erythroid specific gene regulation, a growing list of other genes may also be regulated through the same, or very similar, cis elements in non-erythroid cells. Taken together, these observations argue that comprehensive analysis of the activities of the small Maf proteins may provide a unique perspective for expanding our understanding of transcriptional regulation that can be elicited through interacting transcription factor networks. PMID- 9224594 TI - The role of a basic amino acid cluster in target site selection and non-specific binding of bZIP peptides to DNA. AB - The ability of a transcription factor to locate and bind its cognate DNA site in the presence of closely related sites and a vast array of non-specific DNA is crucial for cell survival. The CREB/ATF family of transcription factors is an important group of basic region leucine zipper (bZIP) proteins that display high affinity for the CRE site and low affinity for the closely related AP-1 site. Members of the CREB/ATF family share in common a cluster of basic amino acids at the N-terminus of their bZIP element. This basic cluster is necessary and sufficient to cause the CRE site to bend upon binding of a CREB/ATF protein. The possibility that DNA bending and CRE/AP-1 specificity were linked in CREB/ATF proteins was investigated using chimeric peptides derived from human CRE-BP1 (a member of the CREB/ATF family) and yeast GCN4, which lacks both a basic cluster and CRE/AP-1 specificity. Gain of function and loss of function experiments demonstrated that the basic cluster was not responsible for the CRE/AP-1 specificity displayed by all characterized CREB/ATF proteins. The basic cluster was, however, responsible for inducing very high affinity for non- specific DNA. It was further shown that basic cluster-containing peptides bind non-specific DNA in a random coil conformation. We postulate that the high non- specific DNA affinities of basic cluster-containing peptides result from cooperative electrostatic interactions with the phosphate backbone that do not require peptide organization. PMID- 9224595 TI - DNA helicase activity in Werner's syndrome gene product synthesized in a baculovirus system. AB - The gene responsible for Werner's syndrome (WRN) contains a region homologous to the Escherichia coli RecQ type DNA helicase and was thought to code for a DNA helicase belonging to this helicase family. However, no evidence has been shown before to substantiate this prediction. Here, we show data that the product of the WRN gene is indeed a DNA helicase. The gene product, a polypeptide with a relative molecular mass of 170 kDa, expressed in the insect Spodoptera frugiperda (Sf21) cell and purified by affinity column chromatography contained both the ATPase and DNA unwinding activities characteristic of DNA helicase. Expressions in Sf21, as well as in HeLa cells, showed that the WRN DNA helicase is exclusively transported to the nucleoplasm, which is consistent with its function in DNA metabolism. Our studies on strand displacement suggest that WRN helicase can unwind not only a duplex DNA, but also an RNA-DNA heteroduplex, while the latter reaction seems less efficient. Enzymological features learned from the purified WRN helicase are discussed with respect to the biological function, which remains to be clarified. PMID- 9224597 TI - Conditional gene targeted deletion by Cre recombinase demonstrates the requirement for the double-strand break repair Mre11 protein in murine embryonic stem cells. AB - Repair of DNA damage resulting in double-strand breaks (DSBs) is controlled by gene products executing homologous recombination or end-joining pathways. The MRE11 gene has previously been implicated in DSB repair in the yeast Saccharomyces cerevisiae . Here we have developed a methodology to study the roles of the murine Mre11 homolog in pluripotent embryonic stem cells. Using a gene targeting approach, a triple LoxP site cassette was inserted into a region of MRE11 genomic DNA flanking conserved phosphodiesterase motifs. The addition of Cre recombinase activity promotes deletions of three types that can be scored. We find that deletion at phosphodiesterase motif III encoded in the N-terminus of Mre11 is acheived in the presence of a wild-type MRE11 allele. However, when the wild-type MRE11 allele is inactivated by gene targeted insertion of a neo marker, only Cre recombination events that allow expression of wild-type Mre11 protein are observed. Therefore, Mre11 is required for normal cell proliferation. This methodology introduces a means to study important regions of essential genes in cell culture models. PMID- 9224596 TI - A branched DNA signal amplification assay for quantification of nucleic acid targets below 100 molecules/ml. AB - The branched DNA hybridization assay has been improved by the inclusion of the novel nucleotides, isoC and isoG, in the amplification sequences to prevent non specific hybridization. The novel isoC, isoG-containing amplification sequences have no detectable interaction with any natural DNA sequence. The control of non specific hybridization in turn permits increased signal amplification. Addition of a 14 site preamplifier was found to increase the signal/noise ratio 8-fold. A set of 74 oligonucleotide probes was designed to the consensus HIV POL sequence. The detection limit of this new HIV branched DNA amplifier assay was approximately 50 molecules/ml. The assay was used to measure viral load in 87 plasma samples of HIV- infected patients on triple drug therapy whose RNA titers were <500 molecules/ml. In all 11 patients viral load eventually declined to below the detection limit with the new assay. PMID- 9224599 TI - Mechanism of DNA transesterification by vaccinia topoisomerase: catalytic contributions of essential residues Arg-130, Gly-132, Tyr-136 and Lys-167. AB - Vaccinia topoisomerase, a eukaryotic type IB enzyme, catalyzes relaxation of supercoiled DNA by cleaving and rejoining DNA strands through a DNA- (3' phosphotyrosyl)-enzyme intermediate. We have performed a kinetic analysis of mutational effects at four essential amino acids: Arg-130, Gly-132, Tyr-136 and Lys-167. Arg-130, Gly-132 and Lys-167 are conserved in all members of the type IB topoisomerase family. Tyr-136 is conserved in all poxvirus topoisomerases. We show that Arg-130 and Lys-167 are required for transesterification chemistry. Arg 130 enhances the rates of both cleavage and religation by 10(5). Lys-167 enhances the cleavage and religation reactions by 10(3) and 10(4), respectively. An instructive distinction between these two essential residues is that Arg-130 cannot be replaced by lysine, whereas substituting Lys-167 by arginine resulted in partial restoration of function relative to the alanine mutant. We propose that both basic residues interact directly with the scissile phosphate at the topoisomerase active site. Mutations at positions Gly-132 and Tyr-136 reduced the rate of strand cleavage by more than two orders of magnitude, but elicited only mild effects on religation rate. Gly-132 and Tyr-136 are suggested to facilitate a pre-cleavage activation step. The results of comprehensive mutagenesis of the vaccinia topoisomerase illuminate mechanistic and structural similarities to site specific recombinases. PMID- 9224598 TI - The developmental activation of the chicken lysozyme locus in transgenic mice requires the interaction of a subset of enhancer elements with the promoter. AB - The complete chicken lysozyme locus is expressed in a position independent fashion in macrophages of transgenic mice and forms the identical chromatin structure as observed with the endogenous gene in chicken cells. Individual lysozyme cis -regulatory elements reorganize their chromatin structure at different developmental stages. Accordingly, their activities are developmentally regulated, indicating a differential role of these elements in locus activation. We have shown previously that a subset of enhancer elements and the promoter are sufficient to activate transcription of the chicken lysozyme gene at the correct developmental stage. Here, we analyzed to which grade the developmentally controlled chromatin reorganizing capacity of cis -regulatory elements in the 5' region of the chicken lysozyme locus is dependent on promoter elements, and we examined whether the lysozyme locus carries a dominant chromatin reorganizing element. To this end we generated transgenic mouse lines carrying constructs with a deletion of the lysozyme promoter. Expression of the transgene in macrophages is abolished, however, the chromatin reorganizing ability of the cis -regulatory elements is differentially impaired. Some cis -elements require the interaction with the promoter to stabilize transcription factor complexes detectable as DNase I hypersensitive sites in chromatin, whereas other elements reorganize their chromatin structure autonomously. PMID- 9224600 TI - A cruciform-dumbbell model for inverted dimer formation mediated by inverted repeats. AB - Small inverted repeats (small palindromes) on plasmids have been shown to mediate a recombinational rearrangement event in Escherichia coli leading to the formation of inverted dimers (giant palindromes). This recombinational rearrangement event is efficient and independent of RecA and RecBCD. In this report, we propose a cruciform-dumbbell model to explain the inverted dimer formation mediated by inverted repeats. In this model, the inverted repeats promote the formation of a DNA cruciform which is processed by an endonuclease into a linear DNA with two hairpin loops at its ends. Upon DNA replication, this linear dumbbell-like DNA is then converted to the inverted dimer. In support of this model, linear dumbbell DNA molecules with unidirectional origin of DNA replication (ColE1 ori ) have been constructed and shown to transform E.coli efficiently resulting in the formation of the inverted dimer. The ability of linear dumbbell DNA to transform E.coli suggests that the terminal loops may be important in bypassing the requirement of DNA supercoiling for initiation of replication of the ColE1 ori. PMID- 9224601 TI - Mechanisms of developmental regulation in Trypanosoma brucei: a polypyrimidine tract in the 3'-untranslated region of a surface protein mRNA affects RNA abundance and translation. AB - Salivarian trypanosomes are extracellular parasites of mammals that are transmitted by tsetse flies. The procyclic acidic repetitive proteins (PARPs) are the major surface glycoproteins of the form of Trypanosoma brucei that replicates in the fly. The abundance of PARP mRNA and protein is very strongly regulated, mostly at the post-transcriptional level. The 3'-untranslated regions of two PARP genes are of similar lengths, but are dissimilar in sequence apart from a 16mer stem-loop that stimulates translation and a 26mer polypyrimidine tract. Addition of either of these PARP 3'-untranslated regions immediately downstream of a reporter gene resulted in developmental regulation mimicking that of PARP. We show that the PARP 3'-UTR reduces RNA stability and translation in bloodstream forms and that the 26mer polypyrimidine tract is necessary for both effects. PMID- 9224602 TI - Synthesis and hybridization properties of inverse oligonucleotides. AB - The synthesis of adenine and thymine cyclopentylethyl nucleosides is presented. This novel constrained monomeric building block is very difficult to incorporate into oligonucleotides. It was introduced in 13mer oligodeoxynucleotide sequences at a single position using H-phosphonate chemistry. Phosphoramidite chemistry completely failed in this particular case. The H-phosphonate building blocks were obtained starting from the corresponding phosphoramidites. Stability of duplexes with RNA and DNA is significantly reduced. PMID- 9224604 TI - Telomere length regulation during postnatal development and ageing in Mus spretus. AB - Telomere shortening has been causally implicated in replicative senescence in humans. To examine the relationship between telomere length and ageing in mice, we have utilized Mus spretus as a model species because it has telomere lengths of approximately the same length as humans. Telomere length and telomerase were analyzed from liver, kidney, spleen, brain and testis from >180 M.spretus male and female mice of different ages. Although telomere lengths for each tissue were heterogeneous, significant changes in telomere lengths were found in spleen and brain, but not in liver, testis or kidney. Telomerase activity was abundant in liver and testis, but weak to non-detectable in spleen, kidney and brain. Gender differences in mean terminal restriction fragment length were discovered in tissues from M.spretus and from M.spretus xC57BL/6 F1 mice, in which a M. spretus -sized telomeric smear could be measured. The comparison of the rank order of tissue telomere lengths within individual M. spretus showed that certain tissues tended to be longer than the others, and this ranking also extended to tissues of the M.spretus xC57BL/6 F1 mice. These data suggest that telomere lengths within individual tissues are regulated independently and are genetically controlled. PMID- 9224603 TI - NMR analysis of CYP1(HAP1) DNA binding domain-CYC1 upstream activation sequence interactions: recognition of a CGG trinucleotide and of an additional thymine 5 bp downstream by the zinc cluster and the N-terminal extremity of the protein. AB - The DNA binding domain of the yeast transcriptional activator CYP1(HAP1) contains a zinc-cluster structure. The structures of the DNA binding domain-DNA complexes of two other zinc-cluster proteins (GAL4 and PPR1) have been studied by X-ray crystallography. Their binding domains present, besides the zinc cluster, a short linker peptide and a dimerization element. They recognize, as homodimers, two rotationally symmetric CGG trinucleotides, the linker peptide and the dimerization element playing a crucial role in binding specificity. Surprisingly, CYP1 recognizes degenerate forms of a direct repeat, CGGnnnTAnCGGnnnTA, and the role of its linker is under discussion. To better understand the binding specificity of CYP1, we have studied, by NMR, the interaction between the CYP1(55 126) peptide and two DNA fragments derived from the CYC1 upstream activation sequence 1B. Our data indicate that CYP1(55-126) interacts with a CGG and with a thymine 5 bp downstream. The CGG trinucleotide is recognized by the zinc cluster in the major groove, as for GAL4 and PPR1, and the thymine is bound in the minor groove by the N-terminal region, which possesses a basic stretch of arginyl and lysyl residues. This suggests that the CYP1(55-126) N-terminal region could play a role in the affinity and/or specificity of the interaction with its DNA targets, in contrast to GAL4 and PPR1. PMID- 9224605 TI - Modification of DNA duplexes to smooth their thermal stability independently of their base content for DNA sequencing by hybridization. AB - The possibility of equalizing DNA duplex stability is essential for the application of sequencing by hybridization. In this paper we describe a new strategy to obtain DNA duplexes with a thermal stability independent of their base content. Modified *C bases have been developed and incorporated into oligonucleotides. The influence of these modifications on duplex stability has been studied by absorption spectroscopy, thus allowing selection of N -4-ethyl-2' deoxycytidine (d4EtC), which hybridizes specifically with natural dG to give a G4EtC base pair whose stability is very close to that of natural AT base pairs. Duplexes built with AT and/or G4EtC base pairs exhibit thermal stabilities independent of their base content in a classical buffer solution, thus enabling control of the stability of DNA hybrids as a function of their length only. PMID- 9224606 TI - Analysis of immunoglobulin Sgamma3 recombination breakpoints by PCR: implications for the mechanism of isotype switching. AB - The molecular mechanism of immunoglobulin switch recombination is poorly understood. Switch recombination occurs between pairs of switch regions located upstream of the constant heavy chain genes. Previously we showed that switch recombination breakpoints cluster to a defined subregion in the Sgamma3, Sgamma1 and Sgamma2b tandem repeats. We have developed a strategy for direct amplification of Smu/Sgamma3 composite fragments as well as Smu and Sgamma3 regions by PCR. This assay has been used to analyze the organization of Smu, Sgamma3 and a series of Smu/Sgamma3 recombination breakpoints from hybridomas and normal mitogen-activated splenic B cells. DNA sequence analysis of the switch fragments showed direct joining of Smu and Sgamma3 without deletions or duplications. Mutations were found in two switch junctions on both sides of the crossover point, suggesting that template switching is the most likely model for the mechanism of switch recombination. Statistical analysis of the positions of the recombination breakpoints in the Sgamma3 tandem repeat indicates the presence of two sub-clusters, suggesting non-random usage of DNA substrate in the recombination reaction. PMID- 9224608 TI - A coalescent approach to the polymerase chain reaction. AB - A versatile algorithm is developed to model PCR on a computer. The method is based on a modification of the coalescent process and provides a general framework to analyse data from PCR. It allows for incorporation of the dynamics of the replication process as described in terms of the number of starting template molecules and cycle-dependent PCR efficiency. The simulation method generates, as a first step, the genealogy of a set of sequences sampled from a final PCR product. In a second step a mutation process is superimposed and the resulting data set is analysed. The efficiency of our algorithm enables us to get reliable approximations of various sample distributions. We demonstrate the relevance of our method with two applications: maximum likelihood estimation of the error rate in PCR and a test of homogeneity of the template. PMID- 9224607 TI - Comparison of the specificities and catalytic activities of hammerhead ribozymes and DNA enzymes with respect to the cleavage of BCR-ABL chimeric L6 (b2a2) mRNA. AB - With the eventual goal of developing a treatment for chronic myelogenous leukemia (CML), attempts have been made to design hammerhead ribozymes that can specifically cleave BCR-ABL fusion mRNA. In the case of L6 BCR-ABL fusion mRNA (b2a2 type; BCR exon 2 is fused to ABL exon 2), which has no effective cleavage sites for conventional hammerhead ribozymes near the BCR-ABL junction, it has proved very difficult to cleave the chimeric mRNA specifically. Several hammerhead ribozymes with relatively long junction-recognition sequences have poor substrate-specificity. Therefore, we explored the possibility of using newly selected DNA enzymes that can cleave RNA molecules with high activity to cleave L6 BCR-ABL fusion (b2a2) mRNA. In contrast to the results with the conventional ribozymes, the newly designed DNA enzymes, having higher flexibility for selection of cleavage sites, were able to cleave this chimeric RNA molecule specifically at sites close to the junction. Cleavage occurred only within the abnormal BCR-ABL mRNA, without any cleavage of the normal ABL or BCR mRNA. Thus, these chemically synthesized DNA enzymes seem to be potentially useful for application in vivo , especially for the treatment of CML, if we can develop exogenous delivery strategies. PMID- 9224609 TI - Ribonucleoprotein formation by the ORF1 protein of the non-LTR retrotransposon Tx1L in Xenopus oocytes. AB - The Tx1L elements constitute a family of site-specific non-LTR retrotransposons found in the genome of the frog Xenopus laevis . The elements have two open reading frames (ORFs) with homology to proteins of retroviruses and other retroelements. This study demonstrates an expected activity of one of the element encoded proteins. The RNA binding properties of ORF1p, the product of the first ORF of Tx1L, were examined after expression from RNA injected into Xenopus oocytes. Using sucrose gradient sedimentation and non-denaturing gel electrophoresis, we show that ORF1p associates with RNA in cytoplasmic ribonucleoprotein (RNP) particles. Discrete RNPs are formed with well-defined mobilities. The ORF1p RNPs are distinct from endogenous RNPs that contain stored oocyte mRNAs and two specific endogenous mRNAs do not become associated with ORF1p. ORF1p appears to be capable of associating with its own mRNA and with other injected RNAs, independent of specific recognition sequences. Although nuclear localization of ORF1p was anticipated, based both on the supposed mechanism of transposition and on the presence of a potential nuclear localization signal, no significant fraction of the protein was found in the oocyte nucleus. Nonetheless, the RNA binding capability of ORF1p is consistent with the proposed model for transposition of non-LTR retrotransposons. PMID- 9224610 TI - Nanoscopic structure of DNA condensed for gene delivery. AB - Scanning force microscopy was used to examine DNA condensates prepared with varying stoichiometries of lipospermine or polyethylenimine in physiological solution. For the first time, individual DNA strands were clearly visualized in incomplete condensates without drying. Using lipospermine at sub-saturating concentrations, discrete nuclei of condensation were observed often surrounded by folded loops of DNA. Similar packing of DNA loops occurred for polyethylenimine induced condensation. Increasing the amount of the condensing agent led to the progressive coalescence or aggregation of initial condensation nuclei through folding rather than winding the DNA. At over-saturating charge ratios of the cationic lipid or polymer to DNA, condensates had sizes smaller than or equal to those measured previously in electron micrographs. Polyethylenimine condensates were more compact than lipospermine condensates and both produced more homogeneously compacted plasmids when used in a 2-4-fold charge excess. The size and morphology of the condensates may affect their efficiency in transfection. PMID- 9224611 TI - Multiplex fluorescence-based primer extension method for quantitative mutation analysis of mitochondrial DNA and its diagnostic application for Alzheimer's disease. AB - A sensitive and highly reproducible multiplexed primer extension assay is described for quantitative mutation analysis of heterogeneous DNA populations. Wild-type and mutant target DNA are simultaneously probed in competitive primer extension reactions using fluorophor-labeled primers and high fidelity, thermostable DNA polymerases in the presence of defined mixtures of deoxy- and dideoxynucleotides. Primers are differentially extended and the resulting products are distinguished by size and dye label. Wild-type:mutant DNA ratios are determined from the fluorescence intensities associated with electrophoretically resolved reaction products. Multiple nucleotide sites can be simultaneously interrogated with uniquely labeled primers of different lengths. The application of this quantitative technique is shown in the analysis of heteroplasmic point mutations in mitochondrial DNA that are associated with Alzheimer's disease. PMID- 9224612 TI - Sp3 encodes multiple proteins that differ in their capacity to stimulate or repress transcription. AB - The product of the retinoblastoma (Rb) susceptibility gene ( RB-1 ) regulates expression of a variety of growth control genes via discrete promoter elements termed retinoblastoma control elements (RCEs). We have previously shown that RCEs are bound and regulated by a common set of ubiquitously expressed nuclear proteins of 115, 95 and 80 kDa, termed retinoblastoma control proteins (RCPs). We have also previously determined that Sp3 and Sp1, two members of the Sp family of transcription factors, encode the 115 and 95 kDa RCPs respectively and that Rb stimulates Sp1/Sp3-mediated transcription in vivo. In this report we have extended these results by determining that the 80 kDa RCP arises from Sp3 mRNA via translational initiation at two internal sites located within the Sp3 trans activation domain. Internally initiated Sp3 proteins readily bind to Sp1 binding sites in vitro yet have little or no capacity to stimulate transcription of Sp regulated genes in vivo. Instead, these Sp3-derived proteins function as potent inhibitors of Sp1/Sp3- mediated transcription. Since cell cycle- or signal- induced expression of a variety of genes, including p21 waf1/cip1, p15 INK4B, CYP11A, mdr1 and acetyl-CoA carboxylase, have been mapped to GC-rich promoter elements that bind Sp family members, we speculate that alterations of the protein and/or DNA binding activities of internally initiated Sp3 isoforms may account in part for the regulation of such differentially expressed genes. PMID- 9224613 TI - Intronic polyadenylation in the human glycinamide ribonucleotide formyltransferase gene. AB - The mouse glycinamide ribonucleotide formyltransferase (GART) locus is known to produce two functional proteins, one by recognition and use of an intronic polyadenylation site and the other by downstream splicing. We now report a similar intronic polyadenylation mechanism for the human GART locus. The human GART gene has two potential polyadenylation signals within the identically located intron as that involved in intronic polyadenylation in the mouse gene. Each of the potential polyadenylation signals in the human gene was followed by an extensive polyT rich tract, but only the downstream signal was preceded by a GT tract. Only the downstream signal was utilized. The polyT rich tract which followed the functional polyadenylation site in the human GART gene was virtually identical in sequence to a similarly placed region in the mouse gene. An exact inverted complement to the polyT rich stretch following the active polyadenylation signal was found in the upstream intron of the human gene, suggesting that a hairpin loop may be involved in this intronic polyadenylation. PMID- 9224614 TI - Analysis of the cleavage reaction of a trans-acting human hepatitis delta virus ribozyme. AB - The cleavage reaction catalyzed by the trans -acting genomic ribozyme of human hepatitis delta virus (HDV) was analyzed with a 13mer substrate (R13) and thio substituted [SR13(Rp) and SR13(Sp)] substrates under single-turnover conditions. The cleavage of RNA by the trans -acting HDV ribozyme proceeded as a first order reaction. The logarithm of the rate of cleavage (kclv) increased linearly (with a slope of approximately 1) between pH 4.0 and 6.0, an indication that a single deprotonation reaction occurred. This result suggests that kclv reflects the rate of the chemical cleavage step, at least around pH 5. The amount of active complex with the SR13(Sp) substrate was almost as large as with R13 (60-80%), whereas the amount of the corresponding active complex formed with the SR13(Rp) substrate was, at most, 20% of this value (with 0.5-100 mM Mg2+ions) at pH 5.0. Nonetheless, the value of kclv for all substrates was almost the same (0.4-0.5 min-1). Neither a 'thio effect' nor a 'Mn2+rescue effect' were observed. These results suggest that Mg2+ions do not interact with pro-R oxygen directly but are essential to the formation of the active complex of the ribozyme and its substrate. PMID- 9224615 TI - An episomal vector for stable tetracycline-regulated gene expression. AB - The recently introduced tetracycline (Tc)-regulatable eukaryotic gene expression system based on the Escherichia coli Tn 10 tetracycline operon has proven to be a powerful tool for controlled expression of a variety of genes in vitro as well as in vivo . Control elements of this expression system are contained in two separate plasmid vectors. The tTA vector encodes a transactivator protein and the tetP vector contains a responsive operator-promoter element (tetP) that controls gene expression depending on tTA binding. Establishment of cell lines expressing a gene of interest under tetP control requires two subsequent rounds of transfection and clonal selection after each transfection. Here we describe a modification of this system in which the tetP element is placed in an episomal EBNA-based plasmid that can be stably maintained in primate but not in rodent cells. Using HeLa and human melanoma cells, we show that upon transient or stable transfection a reporter gene is expressed in a Tc-regulated manner similar to the original system. Thus, this expression system combines the advantages of episomal vectors, such as high efficiency of transfection and time-efficient selection of mass cultures, with tight control of gene expression provided by the Tc regulatable system. PMID- 9224616 TI - Cellular distribution of mammalian DNA topoisomerase II is determined by its catalytically dispensable C-terminal domain. AB - Mammalian cells express two genetically distinct isoforms of DNA topoisomerase II, designated topoisomerase IIalphaand topoisomerase IIbeta. We have recently shown that mouse topoisomerase IIalpha can substitute for the yeast topoisomerase II enzyme and complement yeast top2 mutations. This functional complementation allowed functional analysis of the C-terminal domain (CTD) of mammalian topoisomerase II, where the amino acid sequences are divergent and species specific, in contrast to the highly conserved N-terminal and central domains. Several C-terminal deletion mutants of mouse topoisomerase IIalpha were constructed and expressed in yeast top2 cells. We found that the CTD of topoisomerase IIalphais dispensable for enzymatic activity in vitro but is required for nuclear localization in vivo. Interestingly, the CTD of topoisomerase IIbetawas also able to function as a signal for nuclear targeting. We therefore examined whether the CTD alone is sufficient for nuclear localization in vivo . The C-terminal region was fused to GFP (green fluorescent protein) and expressed under the GAL1 promoter in yeast cells. As expected, GFP signal was exclusively detected in the nucleus, irrespective of the CTD derived from either topoisomerase IIalphaor IIbeta. Surprisingly, when the upstream sequence of each CTD was added nuclear localization of the GFP signal was found to be cell cycle dependent: topoisomerase IIalpha-GFP was seen in the mitotic nucleus but was absent from the interphase nucleus, while topoisomerase IIbeta GFP was detected predominantly in the interphase nucleus and less in the mitotic nucleus. Our results suggest that the catalytically dispensable CTD of topoisomerase II is sufficient as a signal for nuclear localization and that yeast cells can distinguish between the two isoforms of mammalian topoisomerase II, localizing each protein properly. PMID- 9224617 TI - Identification of two novel cis-elements in the promoter of the prostate-specific antigen gene that are required to enhance androgen receptor-mediated transactivation. AB - A monomeric androgen responsive element (ARE) is not sufficient to mediate significant androgen induction of the prostate-specific antigen (PSA) gene. Co transfection experiments using a series of 5'deletion fragments of the proximal promoter region of the PSA gene linked to bacterial chloramphenicol acetyltransferase (CAT) as a reporter have identified two motif sequences which are indispensable for androgen receptor (AR)-mediated transactivation of the PSA promoter and have been designated as motifs A and B respectively. Of note, motif B alone has very little independent enhancer activity regardless of the presence or absence of androgen, whereas multi-copies of motif A exert androgenic inducibility for a heterologous promoter independent of the presence of ARE. Nucleotide substitutions in either motif significantly decrease the androgen inducibility and the nuclear protein binding ability. Furthermore, gel band shift experiments consistently demonstrate that nuclear proteins can bind these motifs, and they are non-receptor factors. Our data indicate that these two DNA motifs are novel cis -regulatory elements and exhibit different mechanisms in cooperation with ARE for AR-mediated transactivation. PMID- 9224618 TI - A branch point consensus from Arabidopsis found by non-circular analysis allows for better prediction of acceptor sites. AB - Little knowledge exists about branch points in plants; it has even been claimed that plant introns lack conserved branch point sequences similar to those found in vertebrate introns. A putative branch point consensus sequence for Arabidopsis thaliana resembling the well known metazoan consensus sequence has been proposed, but this is based on search of sequences similar to those in yeast and metazoa. Here we present a novel consensus sequence found by a non-circular approach. A hidden Markov model with a fixed A nucleotide was trained on sequences upstream of the acceptor site. The consensus found by the Markov model shares features with the metazoan consensus, but differs in its details from the consensus proposed earlier. Despite the fact that branch point consensus sequences in plants are weak, we show that a prediction scheme incorporating them leads to a substantial improvement in the recognition of true acceptor sites; the false positive rate being reduced by a factor of 2. We take this as an indication that the consensus found here is the genuine one and that the branch point does play a role in the proper recognition of the acceptor site in plants. PMID- 9224619 TI - Optimization of the performance of the polymerase chain reaction in silicon-based microstructures. AB - We have demonstrated the ability to perform real-time homogeneous, sequence specific detection of PCR products in silicon microstructures. Optimal design/ processing result in equivalent performance (yield and specificity) for high surface-to-volume silicon structures as compared to larger volume reactions in polypropylene tubes. Amplifications in volumes as small as 0.5 microl and thermal cycling times reduced as much as 5-fold from that of conventional systems have been demonstrated for the microstructures. PMID- 9224620 TI - Identification of an intronic enhancer that nullifies upstream repression of SPARC gene expression. AB - The SPARC gene 5'flanking sequence has been shown to contain enhancer elements, but also negative control elements immediately upstream of the enhancer elements. Although these 5'enhancer elements are active in F9 and PYS-2 cells, their activities are nullified by the 5'repressor activity. In the present study we have identified within intron 1 between nucleotides (nt) +5000 and +5150 of the SPARC gene an enhancer element that bound to two transcription factors of 48 and 52 kDa and between nt +5000 and +5523 a DNase I hypersensitive site. Furthermore, a region containing the 3'intron 1 enhancer element, together with the 5'enhancer elements, neutralized the 5'repressor activity and stimulated efficient transcription. The resulting SPARC promoter activity is about equal in F9, differentiated F9 and PYS-2 cells. We consistently found that the rate of SPARC transcription is nearly the same in F9 and PYS-2 cells. Association of the 3'enhancer element in intron 1 with the DNase I hypersensitive site suggests that both play a role in regulating SPARC expression in vivo . PMID- 9224621 TI - An extracellular matrix response element in the promoter of the LpS1 genes of the sea urchin Lytechinus pictus. AB - The extracellular matrix (ECM) has been shown to play an important role in development and tissue-specific gene expression, yet the mechanism by which genes receive signals from the ECM is poorly understood. The aboral ectoderm-specific LpS1-alpha and -beta genes of Lytechinus pictus , members of the Spec gene family, provide an excellent model system to study ECM- mediated gene regulation. Disruption of the ECM by preventing collagen deposition using the lathrytic agent beta-aminopropionitrile (BAPN) inhibits LpS1 gene transcription. LpS1 transcription resumes after removal of BAPN and subsequent collagen reformation. Using a chloramphenicol acetyltransferase (CAT) reporter gene assay, we show that a 125 bp region of the LpS1-beta promoter from -108 to +17 contains an ECM response element (ECM RE). Insertion of the 125 bp region into the promoter of the metallothionein gene of L. pictus, a gene unaffected by ECM disruption, caused the fused promoter to become ECM dependent. As with the endogenous LpS1 genes, CAT activity directed by the fused LpS1-beta promoter resumed in embryos recovered from ECM disruption. A mutation in a cis -acting element called the proximal G-string, which lies in the 125 bp region, caused CAT activity levels in ECM-disrupted embryos to equal that of the wild-type LpS1-bet apromoter in ECM intact embryos. These results suggest that the intact ECM normally transmits signals to inhibit repressor activity at the proximal G-string in aboral ectoderm cells. Consistent with these results were our findings which showed that in addition to expression in the aboral ectoderm, the proximal G-string mutation caused expression of the CAT gene in oral ectoderm cells. These studies suggested that the proximal G-string serves as a binding site for negative regulation of the LpS1 genes in oral ectoderm during development. We also examined trans acting factors binding the proximal G-string following ECM disruption. Band shift gels revealed a predominant set of slower migrating nuclear proteins from ECM disrupted embryos which bound the proximal G-string. This work suggested that ECM disruption initiates signaling that induces a repressor to bind the ECM RE and/or modifies ECM RE binding proteins, which in turn represses LpS1 gene activity. PMID- 9224622 TI - Magnetic bead capture of cDNAs from double-stranded plasmid cDNA libraries. AB - We have developed a cDNA library screening method which allows the simultaneous screening of >10 ( 12 ) double-stranded plasmid cDNA molecules with minimal a priori sequence knowledge. A biotinylated, gene-specific oligonucleotide probe along with abutting 'blocking' oligos is hybridized to the plasmid cDNA library and the target plasmid retrieved with paramagnetic streptavidin beads and transformed into Escherichia coli. Multiple rounds of enrichment with a target plasmid represented at 0.002-0.0001% resulted in over one-third positive clones. Our method will be useful for isolating even the rarest cDNAs starting from ESTs, isolated exons or homologous sequence information. PMID- 9224624 TI - Measurement of agonist-induced guanine nucleotide turnover by the G-protein Gi1alpha when constrained within an alpha2A-adrenoceptor-Gi1alpha fusion protein. AB - A fusion protein was generated between the porcine alpha2A-adrenoceptor and a pertussis-toxin-insensitive (Cys351-->Gly) variant of the alpha subunit of Gi1alpha by direct in-frame fusion of the N-terminus of the G-protein to the C terminus of the receptor. The fusion protein could be transiently expressed to high levels in COS-7 cells. Addition of the alpha2-adrenoceptor agonist 5-bromo-N (4,5-dihydro- 1H-imidazol-2-yl)-6-quinoxalinamine (UK14304) to membranes of pertussis-toxin-treated transfected cells resulted in a concentration-dependent stimulation of high-affinity GTPase activity. Vmax estimations for the GTPase activity demonstrated an induced catalytic-centre activity of 2.0+/-0.2 min-1 for Gi1alpha when the alpha2A-adrenoceptor was maximally stimulated by UK14304 with a Km for GTP of 0.37+/-0.07 microM. Co-expression of excess beta1gamma2 along with the alpha2A-adrenoceptor-Gi1alpha fusion protein resulted in greater maximal UK14304-induced stimulation of high-affinity GTPase activity (2.1+/-0.2-fold) without alteration in agonist EC50. These studies demonstrate the functionality of the fusion construct, its capacity to interact with betagamma complex and its utility in measuring agonist regulation of the catalytic-centre activity of GTP by a receptor-associated G-protein. PMID- 9224625 TI - Role of ascorbate in the activation of NF-kappaB by tumour necrosis factor-alpha in T-cells. AB - The first product of ascorbate oxidation, the ascorbate free radical (AFR), acts in biological systems mainly as an oxidant, and through its role in the plasma membrane redox system exerts different effects on the cell. We have investigated the role of ascorbate, AFR and dehydroascorbate (DHA) in the activation of the NF kappaB transcription factor in Jurkat T-cells stimulated by tumour necrosis factor-alpha (TNF-alpha). Here we show, by electrophoretic mobility shift assays, that ascorbate increases the binding of NF-kappaB to DNA in TNF-alpha-stimulated Jurkat cells. The ability of ascorbate to enhance cytoplasmic inhibitory IkBalpha protein degradation correlates completely with its capacity to induce NF-kappaB binding to DNA and to potentiate NF-kappaB-mediated transactivation of the HIV-1 long terminal repeat promoter in TNF-alpha-stimulated Jurkat cells but not in cells stimulated with PMA plus ionomycin. AFR behaves like ascorbate, while DHA and ascorbate phosphate do not affect TNF-alpha-mediated NF-kappaB activation. These results provide new evidence for a possible relationship between the activation of the electron-transport system at the plasma membrane by ascorbate or its free radical and redox-dependent gene transcription in T-cells. PMID- 9224626 TI - Identification of an 11-residue portion of CTP-phosphocholine cytidylyltransferase that is required for enzyme-membrane interactions. AB - CTP-phosphocholine cytidylyltransferase (CT) is a key regulatory enzyme in the biosynthesis of phosphatidylcholine (PC) in many cells. Enzyme-membrane interactions appear to play an important role in CT activation. A putative membrane-binding domain appears to be located between residues 236 and 293 from the N-terminus. To map the membrane-binding domain more precisely, glutathione S transferase fusion proteins were prepared that contained deletions of various domains in this putative lipid-binding region. The fusion proteins were assessed for their binding of [3H]PC/oleic acid vesicles. Fusion proteins encompassing residues 267-277 bound to PC/oleic acid vesicles, whereas fragments lacking this region exhibited no specific binding to the lipid vesicles. The membrane-binding characteristics of the CT fusion proteins were also examined using intact lung microsomes. Only fragments encompassing residues 267-277 competed with full length 125I-labelled CT, expressed in recombinant Sf9 insect cells, for microsomal membrane binding. To investigate the role of this region in PC biosynthesis, A549 and L2 cells were transfected with cDNA for CT mutants under the control of a glucocorticoid-inducible long terminal repeat (LTR) promoter. Induction of CT mutants containing residues 267-277 in transfectants resulted in reduced PC synthesis. The decrease in PC synthesis was accompanied by a shift in endogenous CT activity from the particulate to the soluble fraction. Expression of CT mutants lacking this region in A549 and L2 cells did not affect PC formation and subcellular distribution of CT activity. These results suggest that the CT region located between residues 267 and 277 from the N-terminus is required for the interaction of CT with membranes. PMID- 9224623 TI - DNA glycosylases in the base excision repair of DNA. AB - A wide range of cytotoxic and mutagenic DNA bases are removed by different DNA glycosylases, which initiate the base excision repair pathway. DNA glycosylases cleave the N-glycosylic bond between the target base and deoxyribose, thus releasing a free base and leaving an apurinic/apyrimidinic (AP) site. In addition, several DNA glycosylases are bifunctional, since they also display a lyase activity that cleaves the phosphodiester backbone 3' to the AP site generated by the glycosylase activity. Structural data and sequence comparisons have identified common features among many of the DNA glycosylases. Their active sites have a structure that can only bind extrahelical target bases, as observed in the crystal structure of human uracil-DNA glycosylase in a complex with double stranded DNA. Nucleotide flipping is apparently actively facilitated by the enzyme. With bacteriophage T4 endonuclease V, a pyrimidine-dimer glycosylase, the enzyme gains access to the target base by flipping out an adenine opposite to the dimer. A conserved helix-hairpin-helix motif and an invariant Asp residue are found in the active sites of more than 20 monofunctional and bifunctional DNA glycosylases. In bifunctional DNA glycosylases, the conserved Asp is thought to deprotonate a conserved Lys, forming an amine nucleophile. The nucleophile forms a covalent intermediate (Schiff base) with the deoxyribose anomeric carbon and expels the base. Deoxyribose subsequently undergoes several transformations, resulting in strand cleavage and regeneration of the free enzyme. The catalytic mechanism of monofunctional glycosylases does not involve covalent intermediates. Instead the conserved Asp residue may activate a water molecule which acts as the attacking nucleophile. PMID- 9224627 TI - Staurosporine, but not Ro 31-8220, induces interleukin 2 production and synergizes with interleukin 1alpha in EL4 thymoma cells. AB - Protein kinase C (PKC) has been implicated in interleukin 1 (IL1) signal transduction in a number of cellular systems, either as a key event in IL1 action or as a negative regulator. Here we have examined the effects of two PKC inhibitors, staurosporine and the more selective agent Ro 31-8220, on IL1 responses in the murine thymoma line EL4.NOB-1. A 1 h pulse of staurosporine was found to strongly potentiate the induction of IL2 by IL1alpha in these cells. In contrast, neither a pulse nor prolonged incubation with Ro 31-8220 affected the response to IL1alpha. Both agents blocked the response to PMA, however. A 1 h pulse of staurosporine was also found to induce IL2 production on its own, activate the transcription factor nuclear factor kappaB (NFkappaB) and increase the expression of a NFkappaB-linked reporter gene. It synergized with IL1alpha in all of these responses. Ro 31-8220 was again without effect, although both staurosporine and Ro 31-8220 blocked the activation of NFkappaB by PMA. Finally, staurosporine caused the translocation of PKC-alpha and -epsilon, and to a lesser extent PKC-beta, but not PKC-θ or -zeta, from the cytosol to the membrane, although a similar effect was observed with Ro 31-8220. The results suggest that PKC is not involved in IL1alpha signalling in EL4 cells. Furthermore, the potentiating effect of staurosporine on IL1alpha action does not involve PKC inhibition, and is likely to be at the level of NFkappaB activation. PMID- 9224628 TI - A conserved GATA motif in a tissue-specific DNase I hypersensitive site of the cardiac alpha-myosin heavy chain gene. AB - Transgenic analysis has indicated that far upstream regulatory elements of the cardiac alpha-myosin heavy chain (MyHC) gene are required for appropriate transgene expression [Subramaniam, Gulick, Neumann, Knotts and Robbins (1993) J. Biol. Chem. 268, 4331-4336]. In an attempt to identify these as-yet-undefined regulatory elements, we mapped the DNase I hypersensitive sites (DHSs) in the 4 kb upstream region of the hamster cardiac alpha-MyHC gene. When using nuclei isolated from late-gestational and adult heart ventricles, a strong DHS was identified in the -1.9 kb region (alpha-1.9 kb site). It cannot be detected in kidney, liver or cardiofibroblast nuclei. Within this site, we found a conserved GATA-motif that interacts specifically with GATA-binding factors in nuclear extracts of cardiomyocytes at various developmental stages. These data provide further evidence to support the role of GATA factors in the regulation of cardiac alpha-MyHC gene expression. PMID- 9224629 TI - Importance of aspartate-70 in organophosphate inhibition, oxime re-activation and aging of human butyrylcholinesterase. AB - Asp-70 is the defining amino acid in the peripheral anionic site of human butyrylcholinesterase (BuChE), whereas acetylcholinesterase has several additional amino acids, the most important one being Trp-277 (Trp-279 in Torpedo AChE). We studied mutants D70G, D70K and A277W to evaluate the role of Asp-70 and Trp-277 in reactions with organophosphates. We found that Asp-70 was important for binding positively charged echothiophate, but not neutral paraoxon and iso OMPA. Asp-70 was also important for binding of positively charged pralidoxime (2 PAM) and for activation of re-activation by excess 2-PAM. Excess 2-PAM had an effect similar to substrate activation, suggesting the binding of 2 mol of 2-PAM to wild-type but not to the D70G mutant. A surprising result was that Asp-70 was important for irreversible aging, the D70G mutant having a 3- and 8-fold lower rate of aging for paraoxon-inhibited and di-isopropyl fluorophosphate-inhibited BuChE. Mutants of Asp-70 had the same rate constants for phosphorylation and re activation by 2-PAM as wild-type. The A277W mutant behaved like wild-type in all assays. Our results predict that people with the atypical (D70G) variant of BuChE will be more sensitive to the toxic effects of echothiophate, but will be equally sensitive to paraoxon and di-isopropyl fluorophosphate. People with the D70G mutation will be resistant to re-activation of their inhibited BuChE by 2-PAM, but this will be offset by the lower rate of irreversible aging of inhibited BuChE, allowing some regeneration by spontaneous hydrolysis. PMID- 9224630 TI - Expression of stable human O-glycan core 2 beta-1,6-N acetylglucosaminyltransferase in Sf9 insect cells. AB - UDP-GlcNAc:Galbeta1-3GalNAc-R (GlcNAc to GalNAc) beta-1, 6-N acetylglucosaminyltransferase (C2GnT) catalyses the formation of O-glycan core 2. Purification and characterization of C2GnT from natural sources has been hampered by the instability of this enzyme. We have been able to prepare a stable partly purified recombinant human C2GnT by expression of a truncated form of the enzyme in the baculovirus/Spodoptera frugiperda 9 (Sf9) insect cell system. C2GnT activity was secreted into the Sf9 culture medium (15 pmol/min per microl; approx. 0.2 mg/l) and was stable at 4 degrees C either in solution or after lyophilization. Endoglycosidase H and N-glycanase F treatment of the radiolabelled C2GnT indicated the presence of N-glycans at both potential N glycosylation sites. The elimination of one or both of the two potential N glycosylation sites or treatment of the virus-infected insect cells with tunicamycin resulted in loss of enzyme activity due in part to protein degradation. PMID- 9224631 TI - Internalization and down-regulation of the prostacyclin receptor in human platelets. AB - The internalization of [3H]iloprost, a prostacyclin analogue, was studied in human platelets by binding studies. After incubation with [3H]iloprost at 37 degrees C, addition of unlabelled ligand at either 37 degrees C or 4 degrees C caused dissociation of 74% and 52% of the bound ligand respectively, suggesting that a portion had been internalized. The percentage of [3H]iloprost bound at equilibrium to the surface (evaluated by acid treatment) at either 37 degrees C or 4 degrees C was markedly different (80% versus 25%). Internalization was dependent on time and on the ligand nature and concentration. Energy-depleting agents (dinitrophenol and 2-deoxyglucose) completely inhibited internalization, whereas probenecid (inhibitor of organic anion transporters) did not affect it significantly. Subcellular fractionation indicated that, at 4 degrees C or in the absence of ligand, most of the receptor was present in membrane fractions (pellet at 27000 or 105000 g), whereas, when platelets were preincubated at 37 degrees C with iloprost, the receptor was found mainly in the cytosolic fraction. In platelets preincubated with iloprost at 4 degrees C, two classes of binding sites were present, whereas after preincubation at 37 degrees C only the lower-affinity sites were detected. After exposure to the agonist, iloprost-induced inhibition of platelet aggregation and activation of adenylate cyclase and cAMP production were significantly lower. Taken together, these data demonstrate that human platelets can internalize a high-affinity binding site for iloprost, presumably the prostacyclin receptor. PMID- 9224632 TI - Analysis of E-box DNA binding during myeloid differentiation reveals complexes that contain Mad but not Max. AB - It has been shown that during myeloid differentiation the levels of mad1 mRNA are induced before the loss of c-Myc protein. This suggests that inactivation of the differentiation-blocking activity of c-Myc might not occur primarily through the loss of Myc protein, but through an increase in the levels of its antagonist, Mad1. To investigate this question we have analysed the levels of mad1 mRNA during differentiation of myeloid leukaemic HL60 cells. Although levels of mad1 mRNA were moderately increased after induction with phorbol ester, we also found that differentiation could be achieved with other inducers without any concomitant up-regulation of mad1 mRNA. In addition, analysis of E-box DNA binding revealed that, although Myc-Max complexes were lost rapidly after differentiation induction, formation of Mad1-containing complexes only occurred during the later stages of the differentiation programme. Further analysis of these Mad-containing complexes revealed that they were also unlikely to have the capacity to antagonize c-Myc function, as they did not contain Max. Therefore these data suggest that an increase in the levels of mad1 mRNA or the formation of a Mad-Max complex are unlikely to be essential or determining events for myeloid differentiation. In addition, the discovery of DNA-binding complexes that contain Mad1, but not Max, opens up this transcription factor network to include other Max-like proteins or proteins of an unrelated nature. PMID- 9224633 TI - Molecular cloning of up-regulated cytoskeletal genes from regenerating skeletal muscle: potential role of myocyte enhancer factor 2 proteins in the activation of muscle-regeneration-associated genes. AB - A subtractive hybridization and cloning strategy was used to identify genes that are up-regulated in regenerating compared with normal skeletal muscle. The gastrocnemius muscle of CD1 mice was injected with a myotoxic agent (BaCl2). A cDNA library was constructed from the regenerating muscle, and was screened with subtracted probes enriched in genes up-regulated during regeneration. Cofilin and vimentin cDNA clones were isolated. Both cofilin and vimentin were demonstrated to be overexpressed in regenerating compared with non-regenerating muscle (17 fold and 19-fold induction respectively). Cofilin and vimentin mRNAs also exhibited an increased expression in C2C12 myoblasts and a decreased expression in differentiated myotubes. Analysis of the regeneration-induced vimentin enhancer/promoter region revealed a consensus binding site for the myocyte enhancer factor 2 (MEF2) transcription factors. Electrophoretic mobility-shift assays and in vivo reporter assays revealed that MEF2 DNA-binding activity and transcriptional activation are increased in regenerating skeletal muscle, indicating that they may play a role in the activation of muscle genes during regeneration. These data suggest that both cofilin (an actin-regulatory protein) and vimentin (an intermediate filament) may be key components of the cytoskeletal reorganization that mediates muscle cell development and adult skeletal-muscle repair. PMID- 9224634 TI - Nitrogen dioxide depletes uric acid and ascorbic acid but not glutathione from lung lining fluid. AB - The aim of this study was to determine the kinetics of the reactions between the gaseous free-radical pollutant, nitrogen dioxide (NO2), and the water-soluble antioxidants present in respiratory tract lining fluid (RTLF). Samples of RTLF, recovered from 12 subjects (mean age 54.1+/-16.3 years; eight male, four female) as bronchoalveolar lavage (BAL) fluid were exposed ex vivo to NO2 [50-1000 parts per billion (ppb)] for 4 h. For comparison, similar exposures were carried out with single and composite solutions with relevant RTLF antioxidant concentrations. Ascorbic acid (AA), uric acid (UA), GSH depletion, and GSSG and malondialdehyde (MDA) formation were determined with time. In the three models, UA and AA were consumed in a time- and NO2-concentration-related fashion. In addition, their rate of depletion correlated positively with their initial concentration (UA, r=0.92, P<0.05; AA, r=0.94, P<0.05). Little difference was found between the rate of loss of AA (2.2+/-0. 2; 1.9+/-0.5; 1.4+/-0.3 nmol.l-1.h 1.ppb-1), and that of UA (2.4+/-0. 2; 2.1+/-0.6; 1.3+/-0.2 nmol.l-1.h-1.ppb-1) in the three RTLF models examined (single, composite, BAL fluid respectively). GSH loss from BAL fluid (0.2+/-0.1) was significantly less than that seen in either single (1.4+/-0.3) or composite (1.2+/-0.5 nmol.l-1.h-1. ppb-1) antioxidant solutions. In all cases, GSH consumption was significantly less than AA or UA. As model complexity increased, the rate of individual antioxidant loss decreased, such that in BAL fluid, AA, UA and GSH consumption rates were significantly less (P<0. 05) than in the pure or composite antioxidant mixtures. In BAL fluid, little GSSG production was observed at any NO2 concentration. MDA concentration, determined as a measure of lipid peroxidation, did not change following exposure to 50, 150 or 400 ppb NO2, but increased MDA was seen in BAL fluid from 8/12 subjects following exposure to 1000 ppb NO2 for 1 h or more. In conclusion, NO2, at environmentally relevant concentrations, depletes BAL fluid of the antioxidant defences, UA and AA, but not GSH. PMID- 9224635 TI - Tissue-specific expression and promoter analyses of the human tissue kallikrein gene in transgenic mice. AB - The expression of the tissue kallikrein gene is tissue-specific and exhibits a complex pattern of transcriptional and post-translational regulation. Information concerning the mechanism of its tissue-specific expression has been limited owing to the lack of suitable cell lines for the expression study. We approached this problem by introducing human tissue kallikrein gene constructs into mouse embryos, creating transgenic lines carrying its coding sequence with varying lengths of the promoter region. One construct (PHK) contained 801 bp in the 5' flanking region and two deletion constructs contained either 302 bp (D300) or 202 bp (D200) of the promoter region. The expression of human tissue kallikrein in these transgenic mice was monitored by Northern blot, reverse transcriptase-PCR followed by Southern blot, and radioimmunoassay. In all three lines, human tissue kallikrein was expressed predominantly in the pancreas and at lower levels in other tissues, including salivary gland, kidney and spleen. This pattern was similar to that of tissue kallikrein expression in human tissues. The D300 line has higher levels of transgene expression than the D200 and PHK lines. The results indicate that the 202 bp segment immediately upstream of the translation starting site is sufficient to direct a tissue-specific expression pattern of the human tissue kallikrein gene, and that regulatory elements might exist between 801 and -202. PMID- 9224636 TI - Larger increases in sensitivity to paracatalytic inactivation than in catalytic competence during experimental evolution of the second beta-galactosidase of Escherichia coli. AB - Second-order rate constants (M-1.s-1) at 25 degrees C and pH 7.5 for inactivation of first-generation (ebga and ebgb), second-generation (ebgab and ebgabcd) and third-generation (ebgabcde) experimental evolvants of the title enzyme by 2',4' dinitrophenyl 2-deoxy-2-fluoro-beta-D-galactopyranoside are 0.042, 0.30, 10, 24 and 57 respectively. Only partial inactivation is observed, except for ebgabcde. At a single high inactivator concentration, inactivation of the wild-type ebgo is also seen. The changes in sensitivity to the paracatalytic inactivator (over a range of 10(3.3)) are larger than changes in kcat/Km for lactose (over a range of 10(2.7)) or nitrophenyl galactosides (over a range of only 10(1.3)), or changes in degalactosylation rate (over a range of 10(1.7)). These data raise the possibility that evolution in the reverse sense, towards insensitivity to a paracatalytic inactivator with a proportionally lower effect on transformation of substrate, may become a mechanism for the development of bacterial resistance to antibiotics that act by paracatalytic enzyme inactivation. PMID- 9224637 TI - Control of vascular smooth-muscle cell growth by macrophage-colony-stimulating factor. AB - Since in several vascular diseases abnormal vascular smooth-muscle cell (VSMC) proliferation is often associated with the presence of macrophages, we examined whether macrophage-colony-stimulating factor (M-CSF) might play a role in the control of VSMC growth. VSMCs were isolated from rat aorta and maintained in culture. Using a bioassay, a macrophage-colony-stimulating activity was detected in the serum-free supernatant of VSMCs, which could be inhibited by the addition of specific anti-M-CSF antibodies. The presence of M-CSF receptor protein and of M-CSF and M-CSF receptor gene transcripts was demonstrated by immunocytochemistry, using a specific anti-c-Fms antibody and Northern blot analysis respectively. [3H]Thymidine incorporation was measured following the addition to quiescent VSMCs of various dilutions of L929 cell supernatant (as a source of M-CSF) or of recombinant M-CSF. Both exogenous M-CSF and serum-free VSMC conditioned medium promoted DNA synthesis in a concentration-dependent manner, and this effect could be abrogated by the presence of a specific anti-M CSF antibody. Under similar experimental conditions, L929 cell supernatant modulated proto-oncogene expression, as assessed by Northern blot analysis of c fos, c-myc, egr-1 and junB. It was further demonstrated that M-CSF could act in synergy with thrombin, platelet-derived growth factor or basic fibroblast growth factor in promoting VSMC DNA synthesis. These results support the hypothesis that M-CSF affects the growth of cultured rat VSMCs through paracrine/autocrine mechanisms. Its effects at both the macrophage and the VSMC level confer to M-CSF a central role in the development of vascular lesions that occurs during atherosclerotic progression. PMID- 9224638 TI - A Trypanosoma cruzi-secreted 80 kDa proteinase with specificity for human collagen types I and IV. AB - Specific interactions between parasites and extracellular matrix components are an important mechanism in the dissemination of Chagas' disease. Binding of the extracellular matrix proteins to Trypanosoma cruzi receptors has been described as a significant step in this phenomenon. In this study, a specific proteinase activity was identified in cell-free extracts of amastigote, trypomastigote and epimastigote forms of T. cruzi using the collagenase fluorogenic substrate N-Suc Gly-Pro-Leu-Gly-Pro-7-amido-4-methylcoumarin. Isolation of this activity was achieved by a four-step FPLC procedure. Optimal enzyme activity was found to occur at pH 8.0 and was associated with a single T. cruzi 80 kDa protein (Tc 80 proteinase) on SDS/PAGE under reducing conditions. An internal peptide sequence of Tc 80 proteinase was obtained (AGDNYTPPE), and no similarity was found to previously described proteinases of T. cruzi. This enzyme activity is strongly inhibited by HgCl2, tosyl-lysylchloromethane ('TLCK') p-chloromercuribenzoate and benzyloxycarbonyl-Phe-Ala-diazomethane. The purified enzyme was able to hydrolyse purified human [14C]collagen types I and IV at neutral pH, but not 14C-labelled BSA, rat laminin, rabbit IgG or small proteins such as insulin or cytochrome c. In addition, Tc 80 proteinase activity was found to be secreted by T. cruzi forms infective to mammalian cells. Furthermore we demonstrated that purified Tc 80 proteinase mediates native collagen type I hydrolysis in rat mesentery. This feature is compared with that of Clostridium histolyticum collagenase. These findings suggest that Tc 80 proteinase may facilitate T. cruzi host-cell infection by degrading the collagens of the extracellular matrix and could represent a good target for Chagas' disease chemotherapy. PMID- 9224639 TI - Purification and kinetic analysis of pea (Pisum sativum L.) NADPH:protochlorophyllide oxidoreductase expressed as a fusion with maltose binding protein in Escherichia coli. AB - NADPH:protochlorophyllide oxidoreductase (POR) catalyses the light-dependent reduction of protochlorophyllide to chlorophyllide, a key reaction in the chlorophyll biosynthetic pathway. To facilitate structure-function studies, POR from pea (Pisum sativum L.) has been overexpressed in Escherichia coli as a fusion with maltose-binding protein (MBP) at 5-10% of the total soluble cell protein. The fusion protein (MBP-POR) has been purified to greater than 90% homogeneity by a two-step affinity-purification procedure. This represents the first successful overexpression and purification of a plant POR. MBP-POR was found to be active, and the kinetic properties were determined using a continuous assay in which the rate of chlorophyllide formation was measured. The Vmax was 20.6+/-0.9 nmol.min-1.mg-1 and the Km values for NADPH and protochlorophyllide were 8.7+/-1.9 microM and 0.27+/-0.04 microM respectively. These results represent the first determination of the kinetic properties of a pure POR and the first report on the kinetics of POR from a dicotyledenous plant. The experiments described here demonstrate that the enzyme is not a 'suicide' enzyme, and the only components required for catalysis are NADPH, protochlorophyllide and light. Size-exclusion chromatography on a Superose 6 HR column indicated that MBP-POR has a molecular mass of 155 kDa (compared with the molecular mass of 80 kDa estimated by SDS/PAGE), indicating that it behaves as a dimer in solution. This is the first direct determination of the oligomerization state of POR. PMID- 9224640 TI - Responsiveness of human neutrophils to interleukin-4: induction of cytoskeletal rearrangements, de novo protein synthesis and delay of apoptosis. AB - Interleukin-4 (IL-4) and IL-13 are cytokines that share many biological activities. We have previously demonstrated that IL-13 affects a number of neutrophil responses, and here we extend our observations to IL-4. We present, for the first time, direct evidence for the presence of functional IL-4 receptors on human neutrophils. We report that IL-4 induces RNA synthesis in a concentration-dependent manner and, based on observations of the induction of morphological cell shape changes and spreading onto glass, we demonstrate that IL 4 activates neutrophil cytoskeletal rearrangements. We further show that IL-4 is a potent activator of de novo protein synthesis in neutrophils, and we identify by microsequencing one of these proteins as the cytoskeletal protein actin. We were also able to demonstrate for the first time that actin is cleaved into at least two fragments of approximately 30 kDa (pI 5.4) and approximately 25 kDa (pI 5.0) in neutrophils. Finally, we report that IL-4 delays neutrophil apoptosis, as assessed by morphological observations from cytocentrifuge preparations, as well as by measurement of differences in staining by flow cytometry with both propidium iodide and Hoechst reagent. Taken together, we conclude that IL-4 is a more potent neutrophil agonist than previously believed. We discuss the possibility that the induction of the de novo synthesis of actin by IL-4 is related to the mechanism by which this cytokine delays apoptosis; in addition, the cleavage of this protein is likely to contribute to the apoptotic process. PMID- 9224641 TI - Purification and characterization of guinea-pig liver microsomal deacetylase involved in the deacetylation of the O-glucoside of N-hydroxyacetanilide. AB - A microsomal deacetylase that catalyses the deacetylation of the O-glucoside of N hydroxyacetanilide (GHA) was purified from guinea-pig liver. The activity was located exclusively in the microsomes and not detected in the cytosol. The purified GHA deacetylase was a trimeric protein with a molecular mass of 160+/-10 (S.D.) kDa composed of subunits of 53+/-2 kDa; its pI was 4.7. The N-terminal amino acid sequence of GHA deacetylase was similar to those reported for guinea pig and rat liver microsomal carboxylesterases. The GHA deacetylase showed a comparable hydrolytic activity towards p-nitrophenyl acetate (PNPA), although the activities towards N-hydroxyacetanilide, acetanilide and some endogenous acylated compounds were very low or not detectable. The deacetylase activity towards GHA was inhibited by organophosphates but not by p-chloromercuribenzoate, suggesting that GHA deacetylase can be classified as a B-esterase. The enzyme exhibited a positive homotropic co-operativity towards GHA. The values of the Hill coefficient, the half-saturating concentration ([S]0.5) for GHA, and Vmax were 1.59+/-0.03, 5.51+/-0.07 mM and 32.5+/-1.4 micromol/min per mg respectively, at the optimum pH of 8.5. The bell-shaped pH dependence of the Vmax/[S]0.5 profile indicated pKa values attributed to histidine and lysine residues. The study of stoichiometric inhibition by di-isopropyl fluorophosphate and kinetic analysis with the Monod-Wyman-Changeux model suggests that GHA deacetylase has six substrate binding sites and three catalytically essential serine residues per enzyme molecule. PMID- 9224642 TI - Involvement of a region near valine-69 of tissue inhibitor of metalloproteinases (TIMP)-1 in the interaction with matrix metalloproteinase 3 (stromelysin 1). AB - Tissue inhibitors of metalloproteinases (TIMPs) inhibit matrix metalloproteinases (MMPs) by forming a 1:1 stoichiometric complex, but the inhibition mechanism of these inhibitors is not known. Here we have investigated the reactive site of TIMP-1 by its proteinase susceptibility before and after forming a complex with MMP-3 (stromelysin 1). When TIMP-1 was allowed to react with human neutrophil elastase, its inhibitory activity was destroyed. This resulted from cleavage of the Val69-Cys70 bond. However, cleavage of this bond by neutrophil elastase was prevented when TIMP-1 formed a complex with the catalytic domain of MMP-3, and full TIMP-1 activity was restored after dissociation of the complex at pH 3.0 in the presence of EDTA. These results indicate that the region around Val69 closely associates with an active MMP. The three-dimensional structure of the N-terminal domain of TIMP-2 elucidated by NMR studies [Williamson, Martorell, Carr, Murphy, Docherty, Freedman and Feeney (1994) Biochemistry 33, 11745-11759] reveals that Val69 and Cys70 form part of an extended ridge that also includes the N-terminal section of the inhibitor. This region is probably involved in the interaction with the catalytic domains of MMPs. PMID- 9224643 TI - Apolipoprotein E forms stable complexes with recombinant Alzheimer's disease beta amyloid precursor protein. AB - Apolipoprotein E (apoE), a protein genetically linked to the incidence of Alzheimer's disease, forms SDS-stable complexes in vitro with beta-amyloid peptide (Abeta), the primary component of senile plaques. In the present study, we investigated whether apoE was able to bind full-length Abeta precursor protein (APP). Using a maltose-binding-protein-APP fusion protein and human very-low density lipoprotein (VLDL), we detected an interaction of apoE with APP that was inhibited by Abeta or anti-apoE antibody. Saturation-binding experiments indicated a single binding equilibrium with an apparent 1:1 stoichiometry and a dissociation constant of 15 nM. An interaction was also observed using apoE from cerebrospinal fluid or delipidated VLDL, as well as recombinant apoE. APP.apoE complexes were SDS-stable, and their formation was not inhibited by reducing conditions; however, they were dissociated by SDS under reducing conditions. ApoE.APP complexes formed high-molecular-mass aggregates, and competition experiments suggested that amino acids 14-23 of Abeta are responsible for complex formation. Finally, no differences were found when studying the interaction of APP with apoE3 or apoE4. Taken together, our results demonstrate that apoE may form stable complexes with the Abeta moiety of APP with characteristics similar to those of complexes formed with isolated Abeta, and suggest the intriguing possibility that apoE-APP interactions may be pathologically relevant in vivo. PMID- 9224644 TI - Effects of thimerosal on the transient kinetics of inositol 1,4,5-trisphosphate induced Ca2+ release from cerebellar microsomes. AB - Thimerosal, a thiol-reactive reagent, has been shown to increase the cytosolic Ca2+ concentration in a variety of cells by sensitizing inositol 1,4,5 trisphosphate (InsP3) receptors. Thimerosal can have both sensitizing (at concentrations of <2 microM) and inhibitory (at concentrations of >2 microM) effects on InsP3-induced Ca2+ release (IICR) from cerebellar microsomes. Transient kinetic studies were performed by employing a fluorimetric stopped-flow approach using fluo-3. IICR was found to be a bi-exponential process with a fast and a slow component. At a maximal InsP3 concentration (20 microM), the fast phase had a rate constant of 0.9 s-1 and the slow phase had a rate constant of 0.4 s-1. The amplitudes of the two phases were 60% and 40% respectively. When the rate constants for the two phases were plotted as Hill plots, the processes were found to be non-co-operative in both cases (Hill coefficient of 1.0), thus arguing for a simple mechanism linking InsP3 binding to channel opening. At a submaximal InsP3 concentration (0.2 microM), where the sensitizing effects of thimerosal are most pronounced, thimerosal increased the rate constants of both phases in a sigmoidal fashion, with a Hill coefficient of 4.0, suggesting that several cysteine residues (up to four) need to be modified in order for maximum sensitization to occur. The rate constants remained elevated even at thimerosal concentrations that inhibited IICR. The amplitude or extent of Ca2+ release was, however, elevated to a much greater extent in the slow phase, suggesting that the two phases respond differently. At maximal InsP3 concentrations, thimerosal has no effect upon the rate constants but inhibits the amplitude of Ca2+ release. PMID- 9224645 TI - Glutathione and the rate of cellular proliferation determine tumour cell sensitivity to tumour necrosis factor in vivo. AB - Low rates of cellular proliferation are associated with low GSH content and enhanced sensitivity of Ehrlich ascites-tumour (EAT) cells to the cytotoxic effects of recombinant human tumour necrosis factor (rhTNF-alpha). Buthionine sulphoximine, a selective inhibitor of GSH synthesis, inhibited tumour growth and increased rhTNF-alpha cytoxicity in vitro. Administration of sublethal doses (10(6)units/kg per day) of rhTNF-alpha to EAT-bearing mice promoted oxidative stress (as measured by increases in intracellular peroxide levels, O2(-); generation and mitochondrial GSSG) and resulted in a slight reduction (19%) in tumour cell number when controls showed the highest rate of cellular proliferation. ATP (1mmol/kg per day)-induced selective GSH depletion, when combined with rhTNF-alpha administration, afforded a 61% inhibition of tumour growth and resulted in a significant extension of host survival. Administration of N-acetylcysteine (1mmol/kg per day) or GSH ester (5mmol/kg per day) abolished the rhTNF-alpha- and ATP-induced effects on tumour growth by maintaining high GSH levels in the cancer cells. Our results demonstrate that the sensitivity of tumour cells to rhTNF-alpha in vivo depends on their GSH content and their rate of proliferation. PMID- 9224646 TI - Protein involvement in the fusion between the equatorial segment of acrosome reacted human spermatozoa and liposomes. AB - Artificial membranes (liposomes) can interact with the equatorial segment (ES) of human spermatozoa, provided that the acrosome reaction (AR) has occurred [Arts, Kuiken, Jager and Hoekstra (1993) Eur. J. Biochem. 217, 1001-1009]. Using fluorescently labelled liposomes, this interaction can be seen as either punctate fluorescence in the ES (lip-ESp), reflecting only bound liposomes, or as diffuse fluorescence in this region (lip-ESd), indicating that the liposomes have fused with the ES membrane. Only equatorial segments that still contain constituents of the acrosomal matrix have the capacity to bind liposomes and eventually to fuse with them. Since the exposure of such intact equatorial segments is the exclusive result of induction of the AR under physiological conditions, these results imply that liposomes can be used for the rapid detection of acrosome-reacted spermatozoa. The lip-ESp and lip-ESd patterns were shown to be reflections of two distinct properties of the ES. Proteolytic treatment after AR completely inhibited the formation of a lip-ESd pattern, whereas formation of the lip-ESp pattern was only marginally inhibited by the proteolytic treatment. The same results were obtained using anti-sperm antibodies, which did not react with acrosome-intact spermatozoa. Proteolytic treatment of spermatozoa before AR induction had no effect on the fusion capacity of the ES after subsequent AR, which implies that the putative fusion protein is not accessible before AR. Thus fusion of liposomes with the ES of human spermatozoa is mediated by a sperm protein(s), whereas the lip-ESp pattern is not likely to represent the liposome binding stage that precedes the fusion step. PMID- 9224647 TI - Cloning of mouse 17beta-hydroxysteroid dehydrogenase type 2, and analysing expression of the mRNAs for types 1, 2, 3, 4 and 5 in mouse embryos and adult tissues. AB - 17beta-Hydroxysteroid dehydrogenases (17HSDs) are responsible for the conversion of low-activity sex steroids to more potent forms, and vice versa. 17HSD activity is essential for the biosynthesis of sex steroids in the gonads, and it is also one of the key factors regulating the availability of active ligands for sex steroid receptors in various extragonadal tissues. In this study, we have characterized mouse 17HSD type 2 cDNA, and analysed the relative expression of 17HSD types 1, 2, 3, 4 and 5 mRNAs in mouse embryos and adult male and female tissues. The cDNA characterized has a open reading frame of 1146 bp, and encodes a protein of 381 amino acids with a predicted molecular mass of 41837 kDa. Northern-blot analysis of adult mouse tissues revealed that, of the different 17HSDs, the type 2 enzyme is most abundantly expressed. High expression of the enzyme, which oxidizes both testosterone and oestradiol, in several large organs of both sexes indicates that it is the isoform having the most substantial role in the metabolism of sex steroids. Interestingly, four of the five 17HSD enzymes were also detected by Northern blots of whole mouse embryos, and each of the enzymes showed a unique pattern of expression. The oestradiol-synthesizing type 1 enzyme predominates in early days of development embryonic day 7, but after that the oxidative type 2 enzyme becomes the predominant form of all 17HSDs. The data therefore suggest that there is transient oestradiol production in the early days of embryonic development, after which inactivation of sex steroids predominates in the fetus and placenta. PMID- 9224648 TI - Quantitative profiling of tissue- and gender-related expression of glutathione S transferase isoenzymes in the mouse. AB - Cytosolic glutathione S-transferase (GST) isoenzymes from brain, heart, lung, liver, kidney and gonads of male and female CD-1 mice were identified and quantified with a combination of affinity purification, electrospray ionization MS, Edman microsequencing, Western blot analysis and reverse-phase HPLC. The three principal hepatic GST subunits, mGSTA3 (25271 Da), mGSTP1 (23478 Da), and mGSTM1 (25839 Da), were isolated from liver, lung, kidney, testes and female heart, whereas brain, ovaries and male heart contained mGSTM1 and mGSTP1. Additional isoenzymes were detected in tissues, including mu class subunits mGSTM2 (25580 Da) and mGSTM3 (25570 Da), an N-terminally blocked Alpha subunit (25480 Da) assigned as mGSTA4, and proteins of molecular masses 25490, 22540, 24493, 24378 and 25383 Da. Distinct gender differences in expression of GST subunits were observed for liver, heart, kidney and gonads, whereas GST expression was similar in brain and lung for both genders. In contrast with patterns of expression in liver (high ratio of mGSTA3 to mGSTP1 in females relative to males), mGSTP1 was the most abundant subunit in female gonads, whereas mGSTA3 was not present in detectable quantities. The profile of GST expression in kidney was similar to that in liver; however, male kidneys expressed 30% more soluble GST than female kidneys. A striking gender-related difference in GST expression was found in cardiac tissue, where female animals expressed 50% more soluble GST than male tissues, and the GST isoenzyme with the greatest documented activity towards lipid hydroperoxides, mGSTA3, was present in female tissue yet absent from male tissue. These results point to complex gender- and tissue-dependent expression of individual mouse GST isoenzymes. PMID- 9224649 TI - Mitogenic signalling by delta opioid receptors expressed in rat-1 fibroblasts involves activation of the p70s6k/p85s6k S6 kinase. AB - The regulation of mitogenic signalling pathways by G-protein-coupled receptors has been studied in Rat-1 fibroblasts stably transfected with the murine delta opioid receptor. We showed recently that stimulation of this receptor led to the activation of the p42 and p44 isoforms of mitogen-activated protein (MAP) kinase [Burt, Carr, Mullaney, Anderson and Milligan (1996) Biochem. J. 320, 227-235]. The present study has examined the role of the ribosomal S6 kinase p70(s6k) in mitogenic signalling by the delta opioid receptor. Treatment of Rat-1 fibroblasts expressing this receptor with the synthetic enkephalin [d-Ala,d-Leu]-enkephalin (DADLE) led to a dose-dependent increase in p70(s6k) enzyme activity. Activation of p70(s6k) was dependent on the level of delta opioid receptor expressed and was sustained above basal levels for several hours. Immunoblotting revealed that p70(s6k) was subject to increased phosphorylation, the extent of which coincided temporally with enzyme activation. Activation of p70(s6k) by DADLE, but not by platelet-derived growth factor, was blocked by pretreatment of cells with pertussis toxin. Activation of p70(s6k) was also partly blocked by wortmannin, indicating that phosphoinositide 3-OH kinase is required for full activation of p70(s6k) by opioid receptor agonists. Activation of the delta opioid receptor in transfected cells led to increased DNA synthesis. This increase was prevented by rapamycin, which also completely blocked activation of p70(s6k) by DADLE. In addition, prevention of the activation of p42 and p44 MAP kinases also blocked the induction of DNA synthesis by DADLE. These results suggest that the activation of both MAP kinases and p70(s6k) might be crucial to the induction of mitogenic responses by Gi-linked receptors such as the delta opioid receptor. PMID- 9224650 TI - Involvement of diacylglycerol production in activation of nuclear factor kappaB by a CD14-mediated lipopolysaccharide stimulus. AB - Exposure of Chinese hamster CHO-K1 transfectant cells expressing mouse CD14 (CHO/CD14 cells) to lipopolysaccharide (LPS) induced rapid elevation of the cellular diacylglycerol (DAG) and choline/phosphocholine levels and nuclear translocation of nuclear factor kappaB (NFkappaB). When cells were incubated with short-chain DAG analogues or bacterial phospholipase C, NFkappaB activation occurred even without the LPS stimulus. Treatment of CHO/CD14 cells with tricyclo[5.2.1.0(2.6)]decyl-(9[8])xanthogenate (D609), an inhibitor of phosphatidylcholine-specific phospholipase C and phospholipase D, almost completely inhibited not only the LPS-dependent production of DAG and choline/phosphocholine but also the LPS-dependent NFkappaB activation. In contrast, treatment of cells with 1-(6-{[3-methoxyoestra-1,3, 5(10)-trien-17beta yl]-1H-pyrrole-2,5-dione (U73122), an inhibitor of phosphatidylinositol-specific phospholipase C in vitro, did not affect the LPS-dependent activation of NFkappaB. Production of DAG and activation of NFkappaB after the LPS stimulus were observed in mouse macrophage-like J774.1 cells, and this response to LPS by J774. 1 cells was also inhibited by D609. These results suggest that the production of DAG from phosphatidylcholine was upstream of NFkappaB activation in response to a CD14-mediated LPS stimulus. PMID- 9224651 TI - A rapid quantitative assay for the detection of mammalian heparanase activity. AB - Heparan sulphate (HS) is an important component of the extracellular matrix and the vasculature basal laminar which functions as a barrier to the extravasation of metastatic and inflammatory cells. Cleavage of HS by endoglycosidase or heparanase activity produced by invading cells may assist in the disassembly of the extracellular matrix and basal laminar, and thereby facilitate cell migration. Heparanase activity has previously been shown to be related to the metastatic potential of murine and human melanoma cell lines [Nakajima, Irimura and Nicolson (1988) J. Cell. Biochem. 36, 157-167]. To determine heparanase activity, porcine mucosal HS was partially de-N-acetylated and re-N-acetylated with [3H]acetic anhydride to yield a radiolabelled substrate. This procedure prevented the masking of, or possible formation of, new heparanase-sensitive cleavage sites as has been observed with previous methods of radiolabelling. Heparanase activity in a variety of tissues and cell homogenates including human platelets, colonic carcinoma cells, umbilical vein endothelial cells and rat mammary adenocarcinoma cells (both metastatic and non-metastatic variants) and liver homogenates all degraded the substrate in a stepwise fashion from 18.5 to approximately 13, 8 and finally to 4.5 kDa fragments, as assessed by gel filtration analysis, confirming the substrate as suitable for the detection of heparanase activity present in a variety of cells and tissues. A rapid quantitative assay was developed with the HS substrate using a novel method for separating degradation products from the substrate by taking advantage of the decreased affinity of the heparanase-cleaved products for the HS-binding plasma protein chicken histidine-rich glycoprotein (cHRG). Incubation mixtures were applied to cHRG-Sepharose columns, with unbound material corresponding to heparanase-degradation products. Heparanase activity was determined for a variety of human, rat and murine cell and tissue homogenates. The highly metastatic rat mammary adenocarcinoma and murine lung carcinoma cell lines had four to ten times the heparanase activity of non-metastatic variants, confirming the correlation of heparanase activity with metastatic potential. Human cancer patients had twice the serum heparanase levels of normal healthy adults. The assay will be valuable for the determination of heparanase activity from a variety of tissue and cell sources, as a diagnostic tool for the determination of heparanase potential, and for the development of specific inhibitors of heparanase activity and metastasis. PMID- 9224652 TI - Role of sarcoplasmic/endoplasmic-reticulum Ca2+-ATPases in mediating Ca2+ waves and local Ca2+-release microdomains in cultured glia. AB - We have characterized the sarcoplasmic-endoplasmic reticulum Ca2+-ATPase (SERCA) pumps in cultured rat cortical type-1 astrocytes, type-2 astrocytes and oligodendrocytes. Perfusion with 10 microM cyclopiazonic acid (CPA) or 1 microM thapsigargin evoked a large and persistent elevation in cytosolic [Ca2+] in normal Ca2+-containing medium and a small and transient increase in nominally Ca2+-free medium. Subtraction of the response in Ca2+-free medium from that in the control revealed a slow-onset Ca2+-entry response to SERCA inhibition, which began after most of the store depletion had occurred. Thapsigargin- and CPA induced responses propagated as Ca2+ waves, which began in several distinct cellular sites and travelled throughout the cell and through nearby cells, in confluent cultures. Propagation was supported by specialized Ca2+-release sites where the amplitude of the response was significantly higher and the rate of rise steeper. Such higher Ca2+-release kinetics were observed at these sites during Ins(1,4,5)P3-mediated Ca2+ waves in the same cells. Fluorescently tagged thapsigargin labelled SERCA pumps throughout glial cell bodies and processes. In oligodendrocyte processes, multiple domains with elevated SERCA staining were always associated with mitochondria. Our results are consistent with a model in which only a single Ca2+ store, expressing Ins(1,4,5)P3 receptors and SERCAs sensitive to both thapsigargin and CPA, is present in rat cortical glia, and indicate that inhibition of SERCA activates both Ca2+ release as a wavefront and Ca2+ entry via store-operated channels. The spatial relationship between SERCAs and mitochondria is likely to be important for regulating microdomains of elevated Ca2+-release kinetics. PMID- 9224653 TI - Interaction of human neutrophil flavocytochrome b with cytosolic proteins: transferred-NOESY NMR studies of a gp91phox C-terminal peptide bound to p47phox. AB - During activation of the neutrophil NADPH oxidase, cytosolic p47(phox) is translocated to the membrane where it associates with flavocytochrome b via multiple binding regions, including a site in the C-terminus of gp91(phox). To investigate this binding site further, we studied the three-dimensional structure of a gp91(phox) C-terminal peptide (551SNSESGPRGVHFIFNKEN568) bound to p47(phox) using transferred nuclear Overhauser effect spectroscopy (Tr-NOESY) NMR. Using MARDIGRAS analysis and simulated annealing, five similar sets of structures of the p47(phox)-bound peptide were obtained, all containing an extended open bend from Ser5 to Phe14 (corresponding to gp91(phox) residues 555-564). The ends of the peptide were poorly defined, however, suggesting they were more flexible. Therefore further refinement was performed on the Ser5-Phe14 region of the peptide after omitting the ends of the peptide from consideration. In this case, two similar structures were obtained. Both structures again exhibited extended open-bend conformations. In addition, the amino acid side chains that showed evidence of immobilization on binding to p47(phox) correlated directly with those that were found previously to be essential for biological activity. Thus during NADPH oxidase assembly, the C-terminus of gp91(phox) binds to 47(phox) in an extended conformation between gp91(phox) residues 555 and 564, with immobilization of all of the amino acid side chains in the 558RGVHFIF564 region except for His561. PMID- 9224654 TI - Identification and characterization of the MUC2 (human intestinal mucin) gene 5' flanking region: promoter activity in cultured cells. AB - The initiation point for MUC2 gene transcription is located within a 7000-base GC rich region of the mucin gene cluster found on chromosome 11p15.5. The promoter activity of the 5'-flanking region of the MUC2 gene was examined following its cloning into the luciferase-producing pGL2-Basic reporter vector. A short segment comprising bases -91 to -73 relative to the start of transcription was found to be important for basal promoter activity in all cell lines tested. Electrophoretic mobility shift assays demonstrated nuclear protein binding to this region, which contains the consensus CACCC motif (5'-GCCACACCC). This element has been shown to be functionally important in several promoters that are active in diverse cell types. Competition experiments using an Sp1 oligonucleotide and antibody supershift experiments indicated that both Sp1 and other Sp1 family members bind to this element. Inclusion of the region between bases -228 and -171 in pGL2-Basic constructs increased normalized luciferase reporter activity by almost 3-fold in C1a cells, which produce relatively high levels of MUC2 mRNA. Significantly lower levels of normalized luciferase activity resulted when the same construct was transfected into cultured cell lines that express low or undetectable levels of MUC2, suggesting a possible role for this region in conferring cell-type specificity of expression. We also demonstrate, using actinomycin D, that the MUC2 mRNA is long-lived, at least in cultured cells. Moreover, no evidence was found that the MUC2 mRNA turned over more rapidly in LS174T cells, which produce relatively low levels of MUC2 mRNA, as compared with C1a cells, which produce high levels of mRNA. Thus a long mRNA half life appears to be an important mechanism involved in achieving elevated levels of MUC2 mRNA. PMID- 9224655 TI - Alternative splicing of ClC-6 (a member of the CIC chloride-channel family) transcripts generates three truncated isoforms one of which, ClC-6c, is kidney specific. AB - ClC-6 is a protein that structurally belongs to the family of ClC-type chloride channels. We now report the identification of three additional ClC-6 isoforms that are truncated because of alternative splicing. We have isolated, from human K562 cells, four types of ClC-6 cDNAs that encode four distinct ClC-6 protein isoforms. ClC-6a (869 amino acids) corresponds to the previously published ClC-6 protein [Brandt and Jentsch (1995) FEBS Lett. 377, 15-20] and it has a canonical ClC structure. However, ClC-6b (320 amino acids), ClC-6c (353 amino acids) and ClC-6d (308 amino acids) are truncated at their C-termini. Hydropathy-plot analysis indicates that the shortened isoforms contain maximally four (ClC-6b and -6d) or seven (ClC-6c) transmembrane domains. Sequence analysis of a human genomic ClC-6 fragment indicates that the cDNA variability arises from alternative splicing at two different positions: the first alternative site consists of an intron flanked by two alternative donor sites and two alternative acceptor sites, the second being due to an exon that is optionally included or excluded. Reverse-transcription-PCR analysis of ClC-6 expression in human cell lines and tissues shows that the majority (83%) of ClC-6 mRNAs consists of ClC-6a or ClC-6c messengers. Furthermore, in a mouse tissue panel, the ClC-6a mRNA has a relatively broad tissue expression pattern, since it could be detected in brain, kidney, testis, skeletal muscle, thymus and pancreas. In contrast, expression of ClC-6c is more restricted, since it was only detected in kidney. PMID- 9224657 TI - Conditionally immortalized cell lines with differentiated functions established from temperature-sensitive T-antigen transgenic mice. AB - A variety of cell lines with differentiated functions are required to study tissue functions at cellular and molecular levels. Using transgenic mice harbouring ubiquitously expressing the temperature-sensitive T-antigen (ts T antigen) gene of simian virus 40 (SV40), many cell lines were generated. The properties of these established lines suggested that their growth was dependent on T-antigen, and that they retained some of the differentiated functions of each particular tissue. A possible use of these cell lines for tissue functions in vitro is discussed. PMID- 9224656 TI - Expression of the murine RanBP1 and Htf9-c genes is regulated from a shared bidirectional promoter during cell cycle progression. AB - The murine Htf9-a/RanBP1 and Htf9-c genes are divergently transcribed from a bidirectional promoter. The Htf9-a gene encodes the RanBP1 protein, a major partner of the Ran GTPase. The divergently transcribed Htf9-c gene encodes a protein sharing similarity with yeast and bacterial nucleic acid-modifying enzymes. We report here that both mRNA species produced by the Htf9-associated genes are regulated during the cell cycle progression, peak in S phase and decrease during mitosis. Transient expression experiments with reporter constructs showed that cell cycle expression is controlled at the transcriptional level, because the bidirectional Htf9 promoter is down-regulated in growth arrested cells, is activated at the G1/S transition and reaches maximal activity in S phase, though with a different efficiency for each orientation. We have delimited specific promoter regions controlling S phase activity in one or both orientations: identified elements contain recognition sites for members belonging to both the E2F and Sp1 families of transcription factors. Together, the results suggest that the sharing of the regulatory region supports co-regulation of the Htf9-a/RanBP1 and Htf9-c genes in a common window of the cell cycle. PMID- 9224658 TI - Molecular genetic elucidation of the tripartite structure of the Schizosaccharomyces pombe 72 kDa TFIID subunit which contains a WD40 structural motif. AB - BACKGROUND: The multisubunit general transcription factor termed TFIID is comprised of the TATA box DNA binding protein TBP and several TBP-associated factors termed TAFs. Current arguments regarding the mechanisms of regulation of transcription contend that TFIID makes multiple specific protein-protein interactions with numerous protein factors, and that these interactions are important for the regulation of transcriptional initiation. TAFs contain a variety of potential structural motifs and it has been speculated that these motifs participate directly in TAF function. However, to date the physiological significance of these putative structural motifs has not been systematically analysed in vivo. RESULTS: The essential gene encoding the Schizosaccharomyces pombe 72 kDa TFIID subunit is termed taf72+, which contains WD40 repeats, was cloned and sequenced. A comparison of the primary structure of this gene with its Drosophila and S. cerevisiae counterparts suggests the presence of regions that might play a role in TFIID function, due to the fact that significant portions of the sequences are highly conserved. Complementation analyses of a series of deletion mutants of this gene revealed that the most evolutionarily conserved regions of taf72+, including the WD40 repeats, are in fact indispensable for the viability. CONCLUSIONS: The 72 kDa subunit of S. pombe TFIID, which contains putative WD40 repeats, consists of three distinct functional domains separated by intervening regions. The functional significance of the WD40 repeats is demonstrated by this in vivo study. PMID- 9224659 TI - Autoregulation of Pax6 transcriptional activation by two distinct DNA-binding subdomains of the paired domain. AB - BACKGROUND: Pax6 is a transcription factor that plays a central role in eye development. Pax6 contains a DNA-binding domain called paired domain, which consists of a highly conserved N-terminal subdomain and a variable C-terminal subdomain. Recent findings have suggested that both subdomains possess distinct DNA-binding activities. RESULTS: To understand the mechanism of DNA-binding and transcriptional activation by Pax6 via these subdomains, we employed Pax6 paired domain mutants previously isolated from patients with ocular disorders. Analysis of these mutants by gel shift assay revealed that the N-terminal and C-terminal subdomains can independently bind to their respective cognate sites, P6CON and 5aCON. Results from a luciferase assay, however, showed that the two functional subdomains negatively regulate their transactivation potentials each other. Wild type Pax6 and its hyperactive variants show different patterns of DNA contact. CONCLUSION: These results support a new model for the regulation of Pax6 transactivation: When one DNA-binding subdomain binds to its cognate site, the other subdomain also interacts with the flanking sequences nonspecifically, and this interaction constrains its structure to give a reduced level of transactivation. PMID- 9224660 TI - Transcriptional regulation of a Purkinje cell-specific gene through a functional interaction between ROR alpha and RAR. AB - BACKGROUND: The orphan nuclear receptor ROR alpha is highly expressed in the Purkinje cells of the cerebellum during the postnatal development of brain. A recent observation has been made that the ROR alpha gene is disrupted in staggerer mice-which show a cell-autonomous defect in the development of the Purkinje cells. RESULTS: In order to understand the functions of ROR alpha in cerebellar development, I attempted to identify its target genes. Transient expression study demonstrated that transcription of the Purkinje cell protein-2 (Pcp-2) gene is activated by ROR alpha, which binds as a monomer to a single half site motif (RORE) within the promoter region. Its transcription was also activated by retinoic acid receptor (RAR) which binds as a heterodimer with RXR to a retinoic acid responsive element (RARE) in the downstream region. Interestingly, the ROR alpha-mediated transcription is further activated synergistically by RAR. CONCLUSION: That the Pcp-2 gene is a target of ROR alpha, and is suggested that its transcription is also regulated by RAR. PMID- 9224662 TI - Overview of role of BMD measurements in managing osteoporosis. AB - The advent of effective treatments and the opportunity to precisely and accurately measure bone mass have probably been the greatest advances in the field of osteoporosis in the last decade. Bone densitometry has become the most widespread noninvasive method for the detection of osteoporosis and to provide advice on risk of future fractures. It has achieved an unquestioned role in clinical decision making for the management of osteoporotic patients. PMID- 9224661 TI - A novel function of the C-terminal lipid moieties of Rab3A small G protein implicated in Ca2+-dependent exocytosis--inhibition of interaction with GTP and reduction of this inhibition by phospholipid. AB - BACKGROUND: Rab3A small G protein is implicated in Ca2+-dependent exocytosis. It undergoes posttranslational lipid-modifications at its C-terminal region. These lipid moieties are important for the actions of the regulators of Rab3A, but not for the interaction with its downstream target. RESULTS: We have found another function of the C-terminal lipid moieties of Rab3A. GTP rapidly associates with the guanine nucleotide-free form of unmodified Rab3A, but not with the same form of modified Rab3A. Moreover, GTP rapidly dissociates from the GTP-bound form of modified Rab3A, but not from the same form of unmodified Rab3A. The association of GTP with the guanine nucleotide-free form of modified Rab3A is stimulated by the Rab3 GDP/GTP exchange protein (Rab3 GEP), and the dissociation of GTP from the GTP-bound form is markedly reduced by synaptic vesicle phospholipid. CONCLUSIONS: These results suggest that the interaction of the lipid moieties of Rab3A with Rab3 GEP or synaptic vesicles is required for the interaction of modified Rab3A with GTP. Moreover, these results - together with the fact that Rabphilin-3A associated with synaptic vesicles inhibits the activity of Rab3 GTPase-activating protein - suggest that the GTP-bound form of modified Rab3A is associated with synaptic vesicles through both Rabphilin-3A and the vesicle phospholipid. PMID- 9224663 TI - Technical principles of dual energy x-ray absorptiometry. AB - Since its introduction nearly ten years ago, dual-energy x-ray absorptiometry (DXA) has become the single most widely used technique for performing bone densitometry studies. One reason for its popularity is the ability of DXA systems to measure bone mineral density (BMD) in the spine and proximal femur, the two most common sites for osteoporotic fractures. Other advantages of DXA include the exceptionally low radiation dose to patients, short scan times, high resolution images, good precision and inherent stability of calibration. For these reasons DXA scans are widely used to diagnose osteoporosis, assist making decisions in treatment, and as a follow-up response to therapy. Another important application has been the use of DXA in many clinical trials of new treatments for osteoporosis. Since the first generation pencil beam DXA systems became available, the most significant technical innovation has been the introduction of fan beam systems with shorter scan times, increased patient throughput, and improved image quality. New clinical applications include the measurement of lateral spine and total body BMD, body composition, and vertebral morphometry. Despite these advances, posteroanterior (PA) spine and proximal femur scans remain the most widely used application because of their utility in treatment decisions and monitoring response to therapy. PMID- 9224664 TI - Peripheral measurement techniques for the assessment of osteoporosis. AB - Peripheral measurement techniques have been the first to be developed for the assessment of osteoporosis, and they remain useful. Besides traditional approaches such as radiographic absorptiometry (RA), radiogrammetry, and single photon absorptiometry (SPA), new peripheral approaches have been developed that offer powerful ways to assess skeletal status in osteoporosis. These include single x-ray absorptiometry (SXA), peripheral dual x-ray absorptiometry (pDXA), peripheral quantitative computed tomography (pQCT), quantitative ultrasound (QUS) techniques, and magnetic resonance imaging (MRI) approaches. This review describes the current role of peripheral imaging techniques vis-a-vis their central imaging counterparts. Peripheral measurement techniques are attractive because equipment cost is substantially lower, radiation exposure is small, and the devices require less space and sometimes are even portable. Additionally, QUS and MRI offer the potential to measure aspects of bone status beyond the limits of bone densitometry. Peripheral techniques represent important diagnostic methods for the assessment of osteoporosis. PMID- 9224665 TI - Interpretation of bone densitometry studies. AB - Dual-energy x-ray absorptiometry (DXA) is widely used for identifying patients with osteoporosis, making decisions about the commencement of preventive therapy, and following up response to treatment. It is important that radiologists and nuclear medicine physicians issuing clinical reports present clear interpretations that aid the primary care physician in making decisions affecting treatment. This review discusses the principles behind the interpretation of bone mineral density (BMD) studies. After a World Health Organization report published in 1994, osteoporosis is often diagnosed on the basis of the patient's T-score value (difference of BMD from young adult mean normalized to the population SD). T-scores are a measure of current fracture risk. There are problems relating to the use of T-scores in the elderly, and we argue that decisions about treatment are generally best made on the basis of the Z-score value (difference of BMD from age-matched mean normalized to the population SD) because this measures the patient's fracture risk relative to his or her peers. Recent studies confirm that the posteroanterior (PA) projection lumbar spine scan is still the optimum measurement site for monitoring response to treatment. A BMD change of 4.5% is required to register a statistically significant change. PMID- 9224666 TI - Assessment of fracture risk by bone density measurements. AB - Bone densitometry in its various applications has become an established tool for the diagnosis of osteoporosis. Bone density has been shown to be significantly associated with the risk of future fracture in many prospective studies. From long-term prospective studies, it can be concluded that peak bone density and bone loss are important predictors of subsequent fracture, and that fracture can be predicted over a longer period. Bone density predicts fracture even in elderly persons aged 80 years and older. However, in this population some fractures, such as the cervical hip fracture, may be more strongly influenced by other risk factors. The differences between the various densitometric techniques in predicting future osteoporotic fracture of any type is marginal. However, it seems that bone density measurements at the site of fracture do perform better than measurements at other sites. There is no evidence that measuring a second site improves the diagnostic capability of bone densitometry. The association between bone density and future fracture is partly independent of age and other significant predictors of fracture such as falls, cognizance, and mobility. Quantitative ultrasonic measures of bone quality have been shown to have a predictive capability that is comparable to that of bone density. From the perspective that bone densitometry and quantitative ultrasound independently predict fractures, these measures actually seem complementary rather than competitive. Simple geometric measures of the bones such as hip axis length and vertebral depth may be derived from images of bone densitometry scans and are also predictive of hip fracture or vertebral fracture independently of bone density. Using the current knowledge of the association between bone density, quantitative ultrasound, geometric properties, and fractures as well as clinical risk factors, new models for fracture prediction can be developed for future application in clinical practice for the benefit of the individual patient. PMID- 9224667 TI - Vertebral morphometry studies using dual-energy x-ray absorptiometry. AB - Vertebral fractures are one of the most common consequences of osteoporosis. They are usually diagnosed by visual interpretation of lateral radiographs of the lumbar and thoracic spine. Vertebral morphometry, based on measurements of the anterior, middle, and posterior heights of the vertebral bodies from T4 to L4, is a useful adjunct to the visual reading of radiographs. A new generation of dual energy x-ray absorptiometry (DXA) scanners offers software for acquiring lateral images of the spine and performing vertebral morphometry analysis. Advantages of DXA morphometry include straightforward and reproducible patient positioning, absence of geometrical distortion of the image, low radiation dose, digital acquisition, and simplified, semi-automated scan analysis. The widespread availability of such DXA systems should make the investigation of vertebral fractures more widely accessible. PMID- 9224668 TI - Bone scintigraphy in metabolic bone disease. AB - The bone scan has well-recognized appearances in metabolic bone diseases, with its main clinical value found in focal conditions or the focal complications of disease. In clinical practice, the bone scan is most widely used to detect fractures in osteoporosis and pseudofractures in osteomalacia and to evaluate Paget's disease. PMID- 9224669 TI - XATH-1, a vertebrate homolog of Drosophila atonal, induces a neuronal differentiation within ectodermal progenitors. AB - XATH-1, a basic/helix-loop-helix transcription factor and a homolog of Drosophila atonal and mammalian MATH-1, is expressed specifically in the dorsal hindbrain during Xenopus neural development. In order to investigate the role of XATH-1 in the neuronal differentiation process, we have examined the effects of XATH-1 overexpression during Xenopus development. XATH-1 induces the expression of neuronal differentiation markers, such as N-tubulin, within the neural plate as well as within nonneural ectodermal progenitor populations, resulting in the appearance of process-bearing neurons within the epidermis. The related basic/helix-loop-helix genes neurogenin-related-1 and neuroD are not induced in response to XATH-1 overexpression within the embryo, suggesting that XATH-1 may activate an alternate pathway of neuronal differentiation. In further contrast to neurogenin-related-1 and neuroD, high-level expression of general neural markers expressed earlier in development, such as N-CAM, is not induced by XATH-1 overexpression. Competent ectodermal progenitors therefore respond to ectopic XATH-1 expression by initiating a distinct program of neuronal differentiation. PMID- 9224670 TI - Lens regeneration in larval Xenopus laevis: experimental analysis of the decline in the regenerative capacity during development. AB - In Xenopus laevis, the capacity to regenerate a new lens from the outer cornea gradually decreases between stages 50 and 58, is almost negligible during the metamorphic climax, and disappears after metamorphosis. The factors responsible for lens transdifferentiation of the outer cornea are produced by the neural retina and are located in the vitreous chamber. This decrease in the regenerative capacity may be due to: (1) a reduction of the inductive power of the retina, (2) a reduction of lens-forming competence of the outer cornea, (3) an inhibition of the lens transdifferentiation process, (4) a combination of these causes. In order to test these hypotheses, fragments of outer cornea or of outer and inner corneas joined together were isolated from early larvae, late larvae and froglets, and implanted into the eye of host larvae during the premetamorphosis or the metamorphic climax. Results from implants of outer cornea into the vitreous chamber showed that the drop in lens regeneration capacity during the metamorphic climax is not due to a decrease in the inductive power of the retinal factor and that the gradual decrease in the regenerative capacity observed between stages 50 and 58 is not related to a substantial diminution in the capacity of outer cornea cells to transdifferentiate into lens fibers. Results from implants of outer and inner corneas joined together showed that in these implants the lens transdifferentiation of the outer cornea was partially inhibited. These findings indicate that the decrease in lens regeneration is mainly due to an inhibition of the lens transdifferentiation process of the outer cornea by the inner cornea. However, even implants of cornea (multilayered epithelium and substantia propria) excised from metamorphosed animals were able to form lens fibers, although to a lesser percentage than that obtained after implantation of fragments of larval outer and inner corneas. Thus, the lens forming competence in the corneal epithelium is still present to a certain degree even when lens regeneration capacity is lost. Several observations suggest that in the lentectomized eye of late larvae and froglets the mechanical inhibition of lens transdifferentiation process exerted by the inner cornea (or the substantia propria), due to the rapid formation of a connective barrier against the spreading of the retinal factor toward the outer cornea, has a decisive role in maintaining the phenotypic stability of the outer cornea. PMID- 9224671 TI - Sonic hedgehog participates in craniofacial morphogenesis and is down-regulated by teratogenic doses of retinoic acid. AB - The face is one of the most intricately patterned structures in human and yet little is known of the mechanisms by which the tissues are instructed to grow, fuse, and differentiate. We undertook a study to determine if the craniofacial primordia used the same molecular cues that mediate growth and patterning in other embryonic tissues such as the neural tube and the limb. Here we provide evidence for the presence of organizer-like tissues in the craniofacial primordia. These candidate organizers express the polarizing signal sonic hedghog (shh) and its putative receptor, patched, as well as fibroblast growth factor 8 and bone morphogeneic protein 2. Shh-expressing epithelial grafts functioned as organizing tissues in a limb bud assay system, where they evoked duplications of the digit pattern. High doses of retinoic acid, which are known to truncate the growth of the frontonasal and maxillary processes and thus produce bilateral clefting of the lip and palate, inhibited the expression of shh and patched but not fgf8, in the craniofacial primordia, and abolished polarizing activity of these tissues. From these studies we conclude that the embryonic face contains signaling centers in the epithelium that participate in craniofacial growth and patterning. In addition, we discuss a novel mechanism whereby retinoids can exert a teratogenic effect on craniofacial morphogenesis independent of its effects on Hox gene expression or neural crest cell migration. PMID- 9224672 TI - Live astrocytes visualized by green fluorescent protein in transgenic mice. AB - Green fluorescent protein (hGFP-S65T) was expressed in transgenic mice under the control of the astrocyte-specific glial fibrillary acidic protein (GFAP) promoter. Tissues from two independent transgenic lines were characterized by Northern blot analysis and by confocal microscopy. The expression pattern in these two lines was identical in all tissues examined, and similar to that found previously with a lacZ transgene driven by the same promoter. Bright fluorescence was observed in the cell bodies and processes of unfixed or fixed astrocytes, using both whole mount and brain slice preparations, from multiple areas of the central nervous system. However, in contrast to GFAP-lacZ transgenics, retinal Muller cells expressed the GFP transgene in response to degeneration of neighboring photoreceptors. These data indicate that the 2.2-kb hGFAP promoter contains sufficient regulatory elements to direct expression in Muller cells, and that GFP is a suitable reporter gene for use in living preparations of the mammalian nervous system. Such mice should prove useful for studies of dynamic changes in astrocyte morphology during development, and in response to physiological and pathological conditions. PMID- 9224673 TI - Acquisition of meiotic competence in growing mouse oocytes is controlled at both translational and posttranslational levels. AB - Full-grown mouse oocytes spontaneously resume meiosis in vitro when released from their follicular environment. By contrast, growing oocytes are not competent to resume meiosis; the molecular basis of meiotic competence is not known. Entry into M phase of the eukaryotic cell cycle is controlled by MPF, a catalytically active complex comprising p34cdc2 kinase and cyclin B. Incompetent oocytes contain levels of cyclin B comparable to those in competent oocytes, while their level of p34cdc2 is markedly lower; p34cdc2 accumulates abruptly at the end of oocyte growth, at the time of meiotic competence acquisition. We show here that this change in p34cdc2 concentration is not secondary to a corresponding change in the concentration of the cognate mRNA, indicating that translational control may be involved. Microinjection of translatable p34cdc2 mRNA into incompetent oocytes yielded high levels of the protein, but it did not lead to resumption of meiosis. Similarly, microinjection of cyclin B1 mRNA resulted in accumulation of the protein, but not in the acquisition of meiotic competence. By contrast, the microinjection of both p34cdc2 and cyclin B1 mRNAs in incompetent oocytes induced histone H1 and MAP kinase activation, germinal vesicle breakdown, and entry into M-phase including the translational activation of a dormant mRNA. Thus, endogenous cyclin B1 in incompetent oocytes is not available for interaction with p34cdc2, suggesting that a posttranslational event must occur to achieve meiotic competence. Microinjection of either p34cdc2 or cyclin B1 mRNAs accelerated meiotic reinitiation of okadaic acid-treated incompetent oocytes. Taken together, these results suggest that acquisition of meiotic competence by mouse oocytes is regulated at both translational and posttranslational levels. PMID- 9224674 TI - A potential role of R-cadherin in striated muscle formation. AB - We have examined the murine embryonic expression pattern of the cell adhesion molecule R-cadherin in muscle, kidney, thymus, and lung. In developing muscle, R cadherin was first seen at 10.5-11.5 days postcoitum in the somitic myotome. Consistently, we found R-cadherin expressed at the highest levels in the myotome, early skeletal muscle, and smooth muscle (both vascular and visceral), while very low levels of R-cadherin were detected in the heart. The expression pattern and subcellular localization of R-cadherin in developing skeletal muscle indicate a possible role in myoblast cell-cell interactions during both primary and secondary myogenesis. In the developing kidney, R-cadherin was first detected at 10.5 days postcoitum in the mesonephric epithelial tubule cells. In the metanephric kidney, it was specifically expressed in the pretubular aggregates, comma- and S-shaped bodies, proximal tubules, and collecting ducts. Thus, in the kidney, R-cadherin was associated with the mesenchymal-epithelial transition. R cadherin was also found in other developing epithelia, for example in the thymic epithelial cells. In the lung, R-cadherin was expressed at the highest levels in the smooth muscle surrounding the lung epithelial tubules. To test whether R cadherin can direct formation of tissues, we constitutively expressed R-cadherin in E-cadherin-/- ES cells and examined histogenesis in teratomas derived from these cells. R-cadherin exclusively rescued formation of striated muscle and epithelia in the teratomas. R-cadherin's ability to form epithelia in vivo was substantiated by its ability to rescue formation of cystic embryoid bodies in vitro. By comparing our data with the previously reported embryonic expression patterns and histogenetic activities of E- and N-cadherin, we suggest that R cadherin plays an important role in the formation of striated muscle and possibly also of epithelia. PMID- 9224675 TI - Multiple positive cis elements regulate the asymmetric expression of the SpHE gene along the sea urchin embryo animal-vegetal axis. AB - The mechanism that establishes the maternally determined animal-vegetal axis of sea urchin embryos is unknown. We have analyzed the cis-regulatory elements of the SpHE gene of Strongylocentrotus purpuratus, which is asymmetrically expressed along this axis, in an effort to identify components of maternal positional information. Previously, we defined a regulatory region that is sufficient to provide correct nonvegetal expression of a beta-galactosidase reporter gene (Wei, Z., Angerer, L. M., Gagnon, M. L., and Angerer, R. C., Dev. Biol. 171, 195-211, 1995). We have now analyzed this region intensively in order to determine if the spatial pattern is controlled by nonvegetal-positive activities or by vegetal negative activities. The regulatory sequences, except the basal promoter, were mutated by either deletion or sequence replacement. None of these mutations resulted in ectopic beta-gal expression in vegetal cells, showing that no single negative cis element is responsible for the lack of vegetal SpHE transcription. Surprisingly, even short segments of the regulatory region containing only several identified cis elements also direct nonvegetal expression. Furthermore, the SpHE basal promoter functions effectively in vegetal cells in combination with cis-acting elements derived from the PMC-specific gene, SM50. We conclude that the spatial pattern of SpHE transcription is achieved by multiple positive activities concentrated in nonvegetal cells. The vegetal expression of SM50 also is regulated only by positive activities (Makabe, K. W., Kirchhamer, C. V., Britten, R. J., and Davidson, E. H., Development 121, 1957-1970, 1995). A chimeric promoter containing both SpHE and SM50 regulatory sequences is active ubiquitously, suggesting that these regulators are not reciprocally repressive. These observations suggest a model in which the SpHE and SM50 genes are activated by separate sets of positive maternal activities concentrated, respectively, in nonvegetal and vegetal domains of the early embryo. PMID- 9224676 TI - Characterization of the binding of recombinant mouse sperm fertilin beta subunit to mouse eggs: evidence for adhesive activity via an egg beta1 integrin-mediated interaction. AB - The sperm protein fertilin (also known as PH-30) is a candidate for mediating the interactions between sperm and egg plasma membranes. Fertilin is a heterodimer. The beta subunit, which has a region with homology to the family of integrin ligands known as disintegrins, has been hypothesized to be involved in the binding of sperm to the egg surface. To investigate this hypothesis and determine what role fertilin beta plays in fertilization, we have expressed the putative extracellular domain of mouse fertilin beta in bacteria as a fusion protein with maltose-binding protein (hereafter referred to as recombinant fertilin beta-EC) and used two assays to characterize its binding to mouse eggs. Immunocytochemistry was used to examine the localization of recombinant fertilin beta-EC binding. A luminometric assay was also developed to quantify levels of binding of recombinant fertilin beta-EC to single eggs. We find that recombinant fertilin beta-EC binds to the region of the plasma membrane of the egg to which sperm bind, thus providing the first direct evidence that fertilin beta has adhesive properties. Peptides corresponding to the disintegrin domain of fertilin beta reduce its binding to eggs, suggesting that this domain is at least partially involved in the recognition of fertilin beta by binding sites on the egg. Treatment of zona pellucida-free eggs with chymotrypsin reduces the ability of the eggs to support the binding of recombinant fertilin beta-EC, implicating an egg surface protein as a binding site for recombinant fertilin beta-EC. Binding of recombinant fertilin beta-EC to eggs is also reduced in the absence of divalent cations and is supported by 2.0 mM Ca2+, Mg2+, or Mn2+. Furthermore, eggs incubated in recombinant fertilin beta-EC prior to in vitro fertilization show reduced levels of sperm binding. Finally, we have examined the possible role of integrins on eggs as receptors for fertilin beta, since an anti-alpha6 integrin subunit monoclonal antibody, GoH3, has been shown to inhibit sperm binding (E. A. C. Almeida et al. (1995) Cell 81, 1095-1104). We find that: (a) an increased amount of GoH3 epitope on the egg surface does not correlate with an increased ability of the eggs to bind sperm or recombinant fertilin beta-EC; (b) the GoH3 antibody has virtually no inhibitory effect on recombinant fertilin beta EC binding; and (c) recombinant fertilin beta-EC binding is reduced in the presence of anti-beta1 integrin antibodies. These results suggest that a beta1 containing integrin participates in the binding of recombinant fertilin beta-EC to mouse eggs. PMID- 9224677 TI - Characterization of the binding of recombinant mouse sperm fertilin alpha subunit to mouse eggs: evidence for function as a cell adhesion molecule in sperm-egg binding. AB - Fertilin (previously known as PH-30) is a sperm protein that is a candidate molecule for mediating the binding and fusion of the sperm and egg plasma membranes. Fertilin is a heterodimer, with a beta subunit that has a region of homology to the disintegrin family of integrin ligands and an alpha subunit that has a region of homology to viral fusion peptides. It has been hypothesized that fertilin beta and alpha subunits mediate the interactions between sperm and egg plasma membranes, namely, binding and fusion, respectively. To address this hypothesis and to examine specifically the role of fertilin alpha in fertilization, we have expressed the predicted extracellular domain of mouse fertilin alpha as a bacterial fusion protein with maltose-binding protein. This fusion protein (hereafter referred to as recombinant fertilin alpha-EC) binds to the microvillar region of zona pellucida (ZP)-free eggs, the region of the membrane to which sperm bind. This binding is reduced in the absence of divalent cations and is supported by Ca2+, Mg2+, or Mn2+. Eggs that have been treated with chymotrypsin bind less recombinant fertilin alpha-EC than do untreated eggs, suggesting that a chymotrypsin-sensitive binding site for recombinant fertilin alpha-EC is present on egg surfaces. Binding to eggs is also affected by the method used to remove the ZP. Finally, recombinant fertilin alpha-EC inhibits the binding of sperm to eggs during in vitro fertilization of ZP-free eggs. These data are the first evidence to suggest that fertilin alpha can function as a cell adhesion molecule during fertilization, mediating the binding of sperm and egg plasma membranes. PMID- 9224678 TI - Entry of mouse embryonic germ cells into meiosis. AB - Germ cells harvested from mouse embryonic genital ridges were mixed with disaggregated embryonic lung cells, and the reaggregates were cultured for 4-7 days. Germ cells derived from female embryos 10.5-13.5 days postcoitum (dpc) entered and progressed through meiotic prophase in vitro as in vivo, although with a 12- to 24-hr delay. If the cultures were maintained for 2-3 weeks, the germ cells developed into growing oocytes. When germ cells were taken from male embryos 10.5 and 11.5 dpc, they too entered and progressed through meiotic prophase, but germ cells from later embryos (12.5 and 13.5 dpc) developed as prospermatogonia, as in male genital ridges in vivo. When 11.5 dpc male genital ridges were disaggregated, reaggregated, and cultured in the same way as the lung reaggregates, the germ cells again entered meiotic prophase. We conclude that the male genital ridge at about 12 dpc produces a factor that inhibits entry of germ cells into meiosis, and that production of this factor is disrupted by prior disaggregation of the genital ridge. If a meiotic inducing substance is required for entry of germ cells into meiosis, it must be present in the male genital ridge as well as in the female genital ridge, and probably also in the lung. PMID- 9224679 TI - The STUD gene is required for male-specific cytokinesis after telophase II of meiosis in Arabidopsis thaliana. AB - During male meiosis in wild-type Arabidopsis the pollen mother cell (PMC) undergoes two meiotic nuclear divisions in the absence of cell division. Only after telophase II is a wall formed which partitions the PMC into four microspores. Each microspore undergoes two subsequent mitotic divisions to produce one vegetative cell and two sperm cells in the mature pollen grain. In this paper we describe the isolation and the phenotypic characterization of mutations in the STUD (STD) gene, which is specifically required for male specific cytokinesis after telophase II of meiosis. Although the male meiotic nuclear divisions are normal in std mutant plants, no walls are formed resulting in a tetranucleate microspore. Despite the absence of cell division in the PMC, postmeiotic development in the coenocytic microspore proceeds relatively normally, resulting in the formation of large pollen grains which contain four vegetative nuclei and up to eight sperm cells. Interestingly, these enlarged pollen grains which contain multiple vegetative nuclei and extra sperm cells behave as single male gametophytes, producing only single pollen tubes and resulting in partial male fertility in std mutant plants. Characterization of the process of pollen development and pollen function in std mutants thus reveals two different types of developmental regulation. Each of the four nuclei found in a std microspore following meiosis is capable of independently undergoing the complete mitotic cell division (including cytokinesis) which the single nucleus of a wild-type microspore would normally undertake. The ability of the four meiotic products to independently continue through mitosis does not depend on their division into separate cells, but is controlled by some subcellular component found within the coenocytic microspore. By contrast, the mature std pollen grain functions as a unit and produces only a single pollen tube despite the presence of multiple nuclei within the vegetative cell, suggesting that this process is controlled at the cellular level independently of the extra subcellular components. PMID- 9224680 TI - The coupling of cyclic GMP and photopolarization of Pelvetia zygotes. AB - Unidirectional blue light directs the rhizoid-thallus axis in the apolar zygotes of Fucus and Pelvetia. Here, it is shown that blue light (but not red light) increased cyclic GMP levels of Pelvetia zygotes by about a factor of 2. When the increase in cyclic GMP was blocked by a guanylyl cyclase inhibitor, photopolarization was also blocked. Bathing the cells in a permeant cyclic GMP analog, which should tend to collapse intracellular cyclic GMP gradients, reduced the degree of photopolarization. Growing the cells in the dark in a gradient of the analog caused the rhizoids to tend to form on the low concentration side. It appears that the stimulation of the blue light photoreceptors on the side nearer the light activates guanylyl cyclase and results in a transcytoplasmic cyclic GMP gradient that is necessary for polarization. PMID- 9224681 TI - A new model for the membrane topology of glucose-6-phosphatase: the enzyme involved in von Gierke disease. AB - Very recently we have proposed [Hemrika et al. (1997) Proc. Natl. Acad. Sci. USA 94, 2145-2149] that the active site of the vanadate-containing chloroperoxidase from the fungus Curvularia inaequalis, of which the tertiary structure is known, is structurally very similar to that of the membrane-bound mammalian glucose-6 phosphatases for which no structural data are available. The proposed active site of glucose-6-phosphatase, however, is incompatible with the six transmembrane helix topology model that is currently used. Here we present a new topology model for glucose-6-phosphatase which is in agreement with all available data. PMID- 9224682 TI - RNA editing in metazoan mitochondria: staying fit without sex. AB - RNA editing subsumes a number of functionally different mechanisms which have in common that they change the nucleotide sequence of RNA transcripts such that they become different from what would conventionally be predicted from their gene sequences. RNA editing has now been found in the organelles of numerous organisms as well as in a few nuclear transcripts. Most recently, it was shown to affect tRNAs in the mitochondria of several animals. The occurrence and evolutionary persistence of RNA editing is perplexing since backmutations in the genes might be assumed rapidly to eliminate the need for 'correction' of the gene sequences at the post-transcriptional level. Here, we review the recent RNA editing systems discovered in animal mitochondria and propose that they have arisen as a mechanism counteracting the accumulation of mutations that occurs in asexual genetic system. PMID- 9224683 TI - The complete inventory of the yeast Saccharomyces cerevisiae P-type transport ATPases. AB - A total of sixteen open reading frames encoding for P-type ATPases have been identified in the complete genome sequence of Saccharomyces cerevisiae. Phylogenetic analysis distinguishes 6 distinct families. Topology predictions, identification of aminoacid sequence motifs and phenotype analysis of the available mutants suggest that these families correspond to ATPases transporting either H+ (2 members), Ca2+ (2 members), Na+ (3 members), heavy metals (2 members), possibly aminophospholipids (5 members including 4 new ones) or unknown substrates (2 new members). It is proposed that the latter family which has homologs in Tetrahymena thermophila, Plasmodium falciparum and Caenorhabditis elegans constitutes a new group called P4-ATPases with characteristic topology and aminoacid signatures. PMID- 9224684 TI - Addition of G418 and other aminoglycoside antibiotics to mammalian cells results in the release of GPI-anchored proteins. AB - Resistance to the neomycin analogue G418 forms the basis of a dominant marker selection system for mammalian cells transfected with the bacterial neomycin gene. We found that COS-1 cells stably transfected with the neomycin resistance gene had a greater than 50% reduction in cell-associated glycosylphosphatidylinositol (GPI)-anchored alkaline phosphatase (AP). A similarly reduced amount of AP was also observed in wild-type COS-1 cells incubated in the presence of G418 or other aminoglycoside antibiotics. The AP was released from cells into the culture supernatant in its GPI-anchored form. Our data suggest that the G418-induced reduction of AP involves a vesiculation process of COS-1 cells. PMID- 9224685 TI - The interaction of synaptic vesicle-associated membrane protein/synaptobrevin with botulinum neurotoxins D and F. AB - Botulinum neurotoxins type D and F are zinc-endopeptidases with a unique specificity for VAMP/synaptobrevin, an essential component of the exocytosis apparatus. VAMP contains two copies of a nine residue motif, termed V1 and V2, which are determinants of the interaction with tetanus and botulinum B and G neurotoxins. Here, we show that V1 plays a major role in VAMP recognition by botulinum neurotoxins D and F and that V2 is also involved in F binding. Site directed mutagenesis of V1 and V2 indicates that different residues are the determinants of the VAMP interaction with the two endopeptidases. The study of the VAMP-neurotoxins interaction suggest a pairing of the V1 and V2 segments. PMID- 9224686 TI - A single mutation in the M-subunit of Rhodospirillum rubrum confers herbicide resistance. AB - Cells of the photosynthetic bacterium Rhodospirillum rubrum were rendered resistant against the inhibitor 2-(1-phenyl)ethylamino-3-propionylamino-4-cyano thiazole (PPCTH). Electron transport in reaction centers prepared from one of the mutants (M6) was neither inhibited by PPCTH and other NH-thiazoles nor terbutryn. These inhibitors are known to bind at the Q(B) site of the L-subunit. Compared to the wild type, chromatophores from M6 exhibited strongly altered Q(B)- Fe2+ and Q(A)- Fe2+ EPR signals. Inhibitor resistance is due to a mutation in the bacterial reaction center M-subunit, where Glu234 is exchanged against Lys. This is the first example of an inhibitor resistance in the Q(B) site caused by a mutation in the M-subunit. PMID- 9224687 TI - Inhibition of Gal beta1, 4GlcNAc alpha2,6 sialyltransferase expression by antisense-oligodeoxynucleotides. AB - Treatment of human colorectal carcinoma cells HT29 with specific antisense oligodeoxynucleotides led to a decreased Gal beta1,4GlcNAc alpha2,6 sialyltransferase activity on the level of protein expression as well as on the mRNA level. Antisense treatment did not effect cell viability or cell growth. Oligodeoxynucleotides which were complementary to the region upstream of the initiation codon were particularly effective in inhibition of enzyme expression. No such inhibition was found by treatment of cells with oligodeoxynucleotides complementary to the region downstream of the initiation codon or by treatment of cells with scrambled controls or sense oligodeoxynucleotides. PMID- 9224688 TI - Copper and cell-oxidized low-density lipoprotein induces activator protein 1 in fibroblasts, endothelial and smooth muscle cells. AB - The effect of cupric ion- or endothelial cell-oxidized low-density lipoproteins (LDL) on transcription factor AP1 activation was investigated by electrophoretic mobility shift assay. Both oxidized LDL induced AP1 activation in fibroblasts, endothelial and smooth muscle cells. This phenomenon was also observed in the presence of cycloheximide. alpha-Tocopherol, a lipophilic free radical scavenger, and N-acetylcysteine, an hydrophilic antioxidant, partially inhibited the stimulatory effect of Cu2+-oxidized LDL. LDL modified by the mixture of the oxygen radicals OH. and O2.-, which generated lipid peroxidation products, also initiated AP1 activation, whereas LDL modified by OH. alone, which did not lead to marked LDL lipid peroxidation, was ineffective. Thus, lipid peroxidation products seem at least partially involved in the activation mechanism. Since AP1 activity is essential for the regulation of genes involved in cell growth and differentiation, our study suggests that the oxidative stress induced by oxidized LDL might be related to the fibroproliferative response observed in the atherosclerotic plaque. PMID- 9224689 TI - Bacterial aspartic proteinases. AB - Regions of genomic DNA encoding putative aspartic proteinase domains were amplified by PCR from the bacterial species, Escherichia coli and Haemophilus influenzae. Expression of each of these DNA fragments resulted in the accumulation of the corresponding recombinant proteins in insoluble aggregates. Each recombinant protein was solubilised, refolded and shown to be able to cleave synthetic peptides that have been extensively used previously as substrates for aspartic proteinases of vertebrate, fungal and retroviral origin. Each activity was completely blocked by the diagnostic aspartic proteinase inhibitor, acetyl pepstatin. This is thus the first report demonstrating unequivocally that aspartic proteinases may be present in bacteria. PMID- 9224690 TI - Inhibition of the mitochondrial cyclosporin A-sensitive permeability transition pore by the arginine reagent phenylglyoxal. AB - The mitochondrial permeability transition pore, a cyclosporin A-sensitive channel, is controlled by the transmembrane electric potential difference across the inner membrane. Here, we show that treatment of rat liver mitochondria with the arginine reagent phenylglyoxal inhibits the permeability transition pore triggered by depolarization with uncoupler after Ca2+ accumulation. Phenylglyoxal does not change the extent of mitochondrial Ca2+ uptake or the extent of membrane depolarization, indicating that covalent modification of arginine (and possibly lysine) residues directly affects the open probability of the pore. We propose that arginine residues play a role in the physiological control of the permeability transition pore by the mitochondrial transmembrane potential. PMID- 9224691 TI - Resistance to apoptosis in Fanconi's anaemia. An ex vivo study in peripheral blood mononuclear cells. AB - Fanconi's anaemia (FA) is a rare autosomal recessive disease characterised by progressive pancytopoenia, a diverse assortment of congenital malformations, an increased sensitivity to reactive oxygen species and a predisposition to the development of malignancies. In the present study, we assessed the propensity to undergo apoptosis of peripheral blood mononuclear cells (PBMC) from Italian FA patients. Cells were challenged by 2-deoxy-D-ribose (dRib) or TNF-alpha plus cycloheximide as agents that induce apoptosis by interfering with cell redox status and mitochondrial membrane potential (MMP), and PBMC from FA patients resulted to be less prone to die than those from healthy subjects. The decreased susceptibility of FA cells to undergo apoptosis was also evident when another parameter highly correlated with the apoptotic process, i.e. MMP, was measured. Moreover, when N-acetylcysteine was added to dRib-treated PBMC, a strong protection was evident either in PBMC from control subjects or from FA patients. These data indicate that an alteration of unknown nature of the mechanisms favouring apoptosis is present in freshly collected cells from FA patients, and that such alteration could contribute to the pathogenesis of the disease, and particularly to the increased susceptibility to cancer. PMID- 9224692 TI - Substrate and thiol specificity of a stress-inducible glutathione transferase from soybean. AB - An RT-PCR-derived clone encoding a stress-inducible glutathione transferase (GSTGm1) from soybean has been overexpressed in E. coli. The enzyme was active as the dimer GSTGm1-1 and showed GST and glutathione peroxidase activity toward diverse xenobiotics, including analogues of natural stress-metabolites. The selective herbicides, fomesafen and acifluorfen, were conjugated more actively with homoglutathione (hGSH), the major thiol in soybean, than with glutathione (GSH). No thiol preference was shown with the related herbicide, fluorodifen, while GSH was preferred with metolachlor and most non-herbicide substrates. Similar thiol-dependent specificities were observed in GST preparations from plants of varying GSH/hGSH content. PMID- 9224693 TI - A truncated isoform of Ca2+/calmodulin-dependent protein kinase II expressed in human islets of Langerhans may result from trans-splicing. AB - Calcium/calmodulin-dependent protein kinase II (CaM kinase II) has been proposed to play a key role in glucose stimulated insulin secretion. Using the rapid amplification of cDNA ends technique we amplified the 3' end of the CaM kinase II gamma gene from human islet RNA. A novel cDNA was detected composed of 5' sequence from the human CaM kinase II gamma gene joined to the 3' end of the human signal recognition particle 72 (SRP72) gene. We predict that this mRNA species will code for a truncated form of CaM kinase II, designated gammaSRP, comprising the entire catalytic and regulatory domains of the protein and with a predicted molecular weight of 37 kDa. We mapped the human SRP72 gene to chromosome 18 and, as the CaM kinase II gamma gene was previously mapped to human chromosome 10q22, we suggest this novel cDNA may have resulted from trans splicing. PMID- 9224694 TI - Bi-directional movement of actin filaments along long bipolar tracks of oriented rabbit skeletal muscle myosin molecules. AB - In actomyosin in vitro motility assays, orientation of myosin molecules affects their interaction with actin. We obtained long tracks of myosin molecules with uniform orientation. Bipolar filaments about 50 microm long were made from myosin rod prepared from molluscan smooth muscles, to which rabbit skeletal-muscle myosin bound, creating long synthetic thick-filaments. Movement of F-actin toward their center was much faster (4.7 +/- 0.6 microm s(-1)) than in the opposite direction (1.9 +/- 0.2 microm s(-1)), indicating that myosin molecules were arranged in the same orientation along each half of the bipolar filament. These complex thick-filaments permit measurement of actin movement over 20 microm of oriented skeletal myosin tracks facilitating mechanistic studies of actomyosin motility. PMID- 9224695 TI - Thermodynamic characterization of the binding of dCMP to the Asn229Asp mutant of thymidylate synthase. AB - Isothermal titration microcalorimetry and equilibrium dialysis have been used to characterize the binding of 2'-deoxycytidine 5'-monophosphate (dCMP) to the Asn229Asp mutant of Lactobacillus casei recombinant thymidylate synthase at pH 7.4 over a temperature range of 15 degrees C to 35 degrees C. Equilibrium dialysis analysis shows that dCMP binds to two sites in the dimer of both wild type and mutant thymidylate synthase. A concomitant net uptake of protons with binding of dCMP to both enzymes, was detected carrying out calorimetric experiments in various buffer systems with different heats of ionization. The change in protonation for binding of dCMP to wild-type enzyme is lower than that obtained for binding of this nucleotide to TS N229D, which suggests that the pK value of Asp-229 is increased upon dCMP binding to the mutant enzyme. At 25 degrees C, although the binding of dCMP to wild-type and N229D TS is favoured by both enthalpy and entropy changes, the enthalpy change is more negative for the mutant protein. Thus, the substitution of Asn 229 for Asp results in a higher affinity of TS for dCMP due to a more favourable enthalpic contribution. The Gibbs energy change of binding of dCMP to the mutant enzyme is weakly temperature dependent, because of the enthalpy-entropy compensation arising from a negative heat capacity change of binding equal to -0.83 +/- 0.02 kJ K(-1) per mol of dCMP bound. PMID- 9224696 TI - Palytoxin effects through interaction with the Na,K-ATPase in Xenopus oocyte. AB - Palytoxin (PTX) is known to bind to Na,K-ATPase, to inhibit its activity, and to induce cation conductance, but the mechanism of these effects is still poorly understood. In Xenopus oocytes, PTX induced a large cation conductance, an effect that could be prevented or reversed by ouabain for oocytes expressing Xenopus Na,K-pumps but not with those expressing Bufo Na,K-pumps. In both cases patch clamp experiments demonstrated a 7-8 pS channel in the presence of PTX. A large PTX-induced conductance could be observed with minimal Na,K-pump inhibition. From the single PTX-induced channel and macroscopic whole oocyte conductance, and the number of Na,K-pumps, we can conclude that PTX-induced conductance occurs through a direct interaction of PTX with a small number of Na,K-pumps. PMID- 9224697 TI - A novel activity for a group of sesquiterpene lactones: inhibition of aromatase. AB - A group of eleven sesquiterpene lactones isolated from different Asteraceae species from north-western Argentina were investigated for their inhibitory action on the estrogen biosynthesis. Seven of them, of different skeleton types, were found to inhibit the aromatase enzyme activity in human placental microsomes, showing IC50 values ranging from 7 to 110 microM. The most active were the guaianolides 10-epi-8-deoxycumambrin B (compound 1), dehydroleucodin (compound 2) and ludartin (compound 3). These compounds were competitive inhibitors with an apparent Ki = 4 microM, Ki = 21 microM and Ki = 23 microM, respectively. Compounds 1 and 2 acted as type II ligands to the heme iron present in the active site of aromatase cytochrome P450 (P450arom). Besides, all of them failed to affect the cholesterol side-chain cleavage enzyme activity on human placental mitochondrias. This is the first report on the aromatase inhibitory activity of this group of natural compounds. PMID- 9224698 TI - Structural features of the gene encoding human muscle type carnitine palmitoyltransferase I. AB - We isolated a human muscle type of carnitine palmitoyltransferase I (CPTI-M) genomic clone and determined its entire nucleotide sequence. By comparison of the nucleotide sequence of the genomic clone with that of cDNA, we determined the intron/exon junctions. For detection of the exon(s) in the 5'-region of the CPTI M gene, we isolated cDNA clones corresponding to the 5'-region of its transcript by 5'-rapid amplification of cDNA ends (5'-RACE method). Results showed two alternative exons, 1A and 1B, that do not encode amino acids in the 5'-region of the human CPTI-M gene. The gene encoding human CPTI-M was found to consist of two 5'-non-coding exons, 18 coding exons and one 3'-non-coding exon spanning approximately 10 kbp. Furthermore, on analysis of the 5'-flanking region, a putative gene encoding a 'choline kinase homologue' was found to be located only about 300 bp upstream from exon 1A of the human CPTI-M gene. Comparison of the gene structure of human CPTI-M with the reported partial gene structure of human liver type CPTI (CPTI-L) showed that the intron insertion sites were completely conserved in these two genes. PMID- 9224699 TI - Disulfide bond formation is not involved in cap-binding activity of Xenopus translation initiation factor eIF-4E. AB - The eukaryotic initiation factor eIF-4E from Xenopus laevis was expressed in Escherichia coli and refolded in an active form. To define the cysteine residues forming a disulfide bond in Xenopus eIF-4E, each of the 3 cysteine residues was changed to serine by site-directed mutagenesis. Cap-binding activities of the mutant proteins were evaluated by 7-methyl-GTP(m7GTP)-affinity column chromatography. Even the mutant protein containing no cysteine showed an affinity for m7GTP. From the above results and the estimation of the sulfhydryl groups by Ellman's assay method, we concluded that a disulfide bond is not involved in the active Xenopus eIF-4E. PMID- 9224700 TI - Heterogeneity of water-soluble amyloid beta-peptide in Alzheimer's disease and Down's syndrome brains. AB - Water-soluble amyloid beta-peptides (sA beta), ending at residue 42, precede amyloid plaques in Down's syndrome (DS). Here we report that sA beta consists of the full-length A beta(1-42) and peptides truncated and modified by cyclization of the N-terminal glutamates, A beta[3(pE)-42] and A beta[11(pE)-42]. The A beta[3(pE)-42] peptide is the most abundant form of sA beta in Alzheimer's disease (AD) brains. In DS, sA beta[3(pE)-42] concentration increases with age and the peptide becomes a dominant species in the presence of plaques. Both pyroglutamate-modified peptides and the full-length A beta form a stable aggregate that is water soluble. The findings point to a crucial role of the aggregated and modified sA beta in the plaque formation and pathogenesis of AD. PMID- 9224701 TI - Unusual structural stability and ligand induced alterations in oligomerization of a galectin. AB - L-14, a 14-kDa S-type lectin shows the jelly roll tertiary structural fold akin to legume lectins yet, unlike them, it does not dissociate on thermal unfolding. In the absence of ligand L-14 displays denaturation transitions corresponding to tetrameric and octameric entities. The presence of complementary ligand reduces the association of L-14, which is in stark contrast with legume lectins where no alterations in quaternary structures are brought about by saccharides. From the magnitude of the increase in denaturation temperature induced by disaccharides the binding constants calculated from differential scanning calorimetry are comparable with those extrapolated from titration calorimetry indicating that L 14 interacts with ligands essentially in the folded state. PMID- 9224702 TI - Redox chemistry of cobalamin and iron-sulfur cofactors in the tetrachloroethene reductase of Dehalobacter restrictus. AB - Respiration of Dehalobacter restrictus is based on reductive dechlorination of tetrachloroethene. The terminal component of the respiratory chain is the membrane-bound tetrachloroethene reductase. The metal prosthetic groups of the purified enzyme have been studied by optical and EPR spectroscopy. The 60-kDa monomer contains one cobalamin with Em(Co[1+/2+]) = -350 mV and Em(Co[2+/3+]) > 150 mV and two electron-transferring [4Fe-4S](2+;1+) clusters with rather low redox potentials of Em approximately -480 mV. The cob(II)alamin is present in the base-off configuration. A completely reduced enzyme sample reacted very rapidly with tetrachloroethene yielding base-off cob(II)alamin rather than trichlorovinyl cob(III)alamin. PMID- 9224703 TI - A human gene encoding morphine modulating peptides related to NPFF and FMRFamide. AB - FMRFamide-related peptides have been isolated from both invertebrates and vertebrates and exhibit a wide range of biological effects in rats. We show here that in humans 2 FMRFamide-related peptides are encoded by a single gene expressed as a spliced mRNA. The larger predicted peptide (AGEGLNSQFWSLAAPQRFamide) differs from the peptide isolated from bovines (AGEGLSSPFWSLAAPQRFamide) by the substitutions of 2 amino acids. The shorter predicted peptide (NPSF, SQAFLFQPQRFamide) is 3 amino acids longer than the bovine 8 amino-acid NPFF (FLFQPQRFamide) or the human NPFF peptide isolated from serum [5], suggesting that the encoded protein is subject to cleavage by a tripeptidyl peptidase or by a novel processing mechanism. On rat spinal cord, the larger peptide is indistinguishable in activity from the equivalent bovine peptide whereas the smaller extended peptide is inactive. PMID- 9224704 TI - The arrangement of the transmembrane helices in the secretin receptor family of G protein-coupled receptors. AB - The members of the secretin receptor family of G-protein-coupled receptors share no significant sequence similarity to the more familiar rhodopsin-like family. However, multiple sequence alignment analysis reveals seven hydrophobic regions with significant alpha-helical periodicity. Residues that are likely to be buried on the interior of the helical bundle and others that are likely to contact the lipid bilayer are identified. A predicted arrangement of the helical bundle is described in which, by comparison with the arrangement in the rhodopsin family, helices 2 and 7 are more buried within the bundle while helix 3 is more exposed to the lipid bilayer. PMID- 9224705 TI - Triabodies: single chain Fv fragments without a linker form trivalent trimers. AB - A single chain Fv fragment (scFv) of the murine monoclonal antibody 11-1G10 was constructed by directly joining the C-terminal residue of the V(H) domain to the N-terminal residue of V(L). 11-1G10 is an anti-idiotype and competes with the antigen, influenza virus neuraminidase (NA), for binding to the NC41 antibody. The scFv formed stable trimers with three active antigen combining sites for NC41 Fab fragments. We propose that trimeric scFvs may be the preferred conformation for directly linked V(H)-V(L) molecules, which contrasts the formation of scFv dimers (diabodies) when the V(H) and V(L) domains are joined by short flexible linkers of between 5-10 residues. BIAcore biosensor binding experiments showed that the trimeric scFv showed an expected increase in binding affinity, due to avidity, compared to the monomeric 15-residue linked scFv. The increase in avidity of scFv trimers offers advantages for imaging and immunotherapy. PMID- 9224706 TI - Expression of trypsin in vascular endothelial cells. AB - Proteinases produced by vascular endothelial cells are expected to play important roles in many biological processes. Here we report that human vascular endothelial cells express trypsinogen-2 mRNA and its protein product in culture. The trypsinogen production was stimulated by a tumor promoter and associated with cell growth. In situ hybridization analysis showed that the trypsinogen gene was significantly expressed in vascular endothelial cells around gastric tumors and in patients with disseminated intravascular coagulation (DIC). These results suggest the possible roles of endothelial cell-derived trypsin in tumor angiogenesis and abnormal blood coagulation. PMID- 9224707 TI - UDP-galactose 4-epimerase from Escherichia coli: existence of a catalytic monomer. AB - UDP-galactose 4-epimerase from Escherichia coli is a homodimer of molecular mass 39 kDa/subunit and requires NAD as a co-factor. X-ray crystallographic studies indicate two pyridine nucleotide co-factor-binding sites of the dimeric molecule situated in a symmetry-oriented manner. Size-exclusion HPLC of an equilibrium intermediate at 3 M urea suggests a monomeric holoenzyme structure that is catalytically active. Ultracentrifugal studies of the native enzyme in a 5-20% sucrose gradient at low protein concentration also indicate existence of a catalytic monomer. The monomer resembles the dimeric protein in stability and most of its physico-chemical properties. PMID- 9224709 TI - Phosphate groups in lipopolysaccharides of Salmonella typhimurium rfaP mutants. AB - Lipopolysaccharides (LPS) of Salmonella typhimurium rfaP mutants and of a galE strain as a control were subjected to analysis by 31P-NMR in order to assess the location of phosphate groups. This was done to obtain direct proof for our earlier finding by chemical analysis that phosphate was lacking in the core oligosaccharide part of the mutant LPS, whereas the core oligosaccharide normally contains several phosphate groups. Such phosphate deficiency has been associated with the increased susceptibility of the rfaP mutants to hydrophobic antibiotics and detergents. Analysis of the de-O-acylated LPS derivatives of S. typhimurium rfaP strains SH7770, SH8551, and SH8572 by 31P-NMR revealed an almost total lack of phosphate groups in the core oligosaccharide part, the LPS phosphates being largely accounted for by the two monophosphate monoesters of lipid A, linked to positions C-1 and C-4' of the lipid A backbone. Core oligosaccharide-linked phosphates were detected in minor proportions only, indicating the presence of some normally phosphorylated core oligosaccharide, due to the inherently leaky nature of the mutation. PMID- 9224708 TI - AMP-activated protein kinase isoenzyme family: subunit structure and chromosomal location. AB - The AMP-activated protein kinase (AMPK) consists of catalytic alpha and non catalytic, beta and gamma (38 kDa) subunits and is responsible for acting as a metabolic sensor for AMP levels. There are multiple genes for each subunit and we find that rat liver AMPK-alpha2 isoform catalytic subunit is associated with beta1 and gamma1 and not with beta2 or gamma2 subunit isoforms. The beta1 and gamma1 isoforms are also subunits of the alpha1 isoform. The sequence of cloned human AMPK-beta1 is 95% identical in amino acid sequence with rat beta1. Human chromosomal localizations were determined for AMPK-alpha1 (5p11-p14), AMPK-beta1 (12q24.1-24.3) and AMPK-gamma1 (12q12-q14), respectively. PMID- 9224710 TI - Vp165 and GLUT4 share similar vesicle pools along their trafficking pathways in rat adipose cells. AB - vp165 (or gp160) is an aminopeptidase that has been identified as one of the major proteins of the GLUT4-containing vesicles. In the present study we have determined the degree of co-localization between vp165 and GLUT4 in rat adipose cells and used perturbation by wortmannin to assess the exocytic and endocytic steps along the translocation and recycling pathways of GLUT4 in the absence and presence of insulin. Western blots of subcellular membrane fractions demonstrate very similar distributions of vp165 and GLUT4. Confocal microscopy of whole cells provides direct evidence that these proteins share the same vesicle populations moving both towards and from the plasma membrane. These data are consistent with the presence of a distinct insulin-sensitive compartment that sequesters both GLUT4 and vp165 and suggest similar trafficking routes through the recycling compartments. PMID- 9224711 TI - Constitutive activation of the TSH receptor by spontaneous mutations affecting the N-terminal extracellular domain. AB - Activating mutations of the TSH receptor gene have been found in toxic adenomas and hereditary toxic thyroid hyperplasia. Up to now, all mutations have been located in the serpentine portion of the receptor. We now describe two additional mutations affecting Ser-281 (Ser-281-Thr and Ser-281-Asn) in the ectodomain of the receptor. After transfection in COS cells, both mutants displayed increased constitutive activity for cAMP generation despite expression at a lower level than the wild type. The mutants were responsive to TSH. The present results are compatible with a model in which the activity of the unliganded receptor is kept at a low level by an inhibitory interaction between the N-terminal domain and the serpentine portion of the receptor. PMID- 9224712 TI - The Na+/e- stoichiometry of the Na+-motive NADH:quinone oxidoreductase in Vibrio alginolyticus. AB - A method is proposed to estimate the stoichiometries of primary Na+-pumps in intact bacterial cells. It is based on technique when the H+/e- stoichiometry is measured in the presence of protophorous uncoupler and in the absence of penetrating ions other than H+. Under these conditions, the H+ influx discharges membrane potential generated by the Na+ pump so the Na+/e- and H+/e- ratios become equal. Using this approach it is shown that the Na+/e- ratio for the Na+ motive NADH:quinone oxidoreductase of Vibrio alginolyticus is equal to 0.71 +/- 0.06. The Na+/e- stoichiometry appears to be approximately 1, provided that the contribution of the non-coupled NADH:quinone oxidoreductase, which is resistant to low HQNO concentrations, is taken into account. PMID- 9224713 TI - Signal perception and transduction in plant defense responses. PMID- 9224714 TI - Yeast Gcn5 functions in two multisubunit complexes to acetylate nucleosomal histones: characterization of an Ada complex and the SAGA (Spt/Ada) complex. AB - The transcriptional adaptor protein Gcn5 has been identified as a nuclear histone acetyltransferase (HAT). Although recombinant yeast Gcn5 efficiently acetylates free histones, it fails to acetylate histones contained in nucleosomes, indicating that additional components are required for acetylation of chromosomal histones. We report here that Gcn5 functions as a catalytic subunit in two high molecular-mass native HAT complexes, with apparent molecular masses of 0.8 and 1.8 megadalton (MD), respectively, which acetylate nucleosomal histones. Both the 0.8- and 1.8-MD Gcn5-containing complexes cofractionate with Ada2 and are lost in gcn5delta, ada2delta, or ada3delta yeast strains, illustrating that these HAT complexes are bona fide native Ada-transcriptional adaptor complexes. Importantly, the 1.8-MD adaptor/HAT complex also contains Spt gene products that are linked to TATA-binding protein (TBP) function. This complex is lost in spt20/ada5delta and spt7delta strains and Spt3, Spt7, Spt20/Ada5, Ada2, and Gcn5 all copurify with this nucleosomal HAT complex. Therefore, the 1.8-MD adaptor/HAT complex illustrates an interaction between Ada and Spt gene products and confirms the existence of a complex containing the TBP group of Spt proteins as demonstrated by genetic and biochemical studies. We have named this novel transcription regulatory complex SAGA (Spt-Ada-Gcn5-Acetyltransferase). The function of Gcn5 as a histone acetyltransferase within the Ada and SAGA adaptor complexes indicates the importance of histone acetylation during steps in transcription activation mediated by interactions with transcription activators and general transcription factors (i.e., TBP). PMID- 9224715 TI - ELT-1, a GATA-like transcription factor, is required for epidermal cell fates in Caenorhabditis elegans embryos. AB - Epidermal cells are generated during Caenorhabditis elegans embryogenesis by several distinct lineage patterns. These patterns are controlled by maternal genes that determine the identities of early embryonic blastomeres. We show that the embryonically expressed gene elt-1, which was shown previously to encode a GATA-like transcription factor, is required for the production of epidermal cells by each of these lineages. Depending on their lineage history, cells that become epidermal in wild-type embryos become either neurons or muscle cells in elt-1 mutant embryos. The ELT-1 protein is expressed in epidermal cells and in their precursors. We propose that elt-1 functions at an early step in the specification of epidermal cell fates. PMID- 9224716 TI - Genetic analysis reveals that PAX6 is required for normal transcription of pancreatic hormone genes and islet development. AB - We present genetic and biochemical evidence that PAX6 is a key regulator of pancreatic islet hormone gene transcription and is required for normal islet development. In embryos homozygous for a mutant allele of the Pax6 gene, Small eye (Sey(Neu)), the numbers of all four types of endocrine cells in the pancreas are decreased significantly, and islet morphology is abnormal. In the remaining islet cells, hormone production, particularly glucagon production, is markedly reduced because of decreased gene transcription. These effects appear to result from a lack of PAX6 protein in the mutant embryos. Biochemical studies identify wild-type PAX6 protein as the transcription factor that binds to a common element in the glucagon, insulin, and somatostatin promoters, and show that PAX6 transactivates the glucagon and insulin promoters. PMID- 9224717 TI - p21CIP1-mediated inhibition of cell proliferation by overexpression of the gax homeodomain gene. AB - gax, a diverged homeobox gene expressed in vascular smooth muscle cells (VSMCs), is down-regulated in vitro by mitogen stimulation and in vivo in response to vascular injury that leads to cellular proliferation. Recombinant Gax protein microinjected into VSMCs and fibroblasts inhibited the mitogen-induced entry into S-phase when introduced either during quiescence or early stages of G1. Overexpression of gax with a replication-defective adenovirus vector resulted in G0/G1 cell cycle arrest of VSMCs and fibroblasts. The gax-induced growth inhibition correlated with a p53-independent up-regulation of the cyclin dependent kinase inhibitor p21. Gax overexpression also led to an association of p21 with cdk2 complexes and a decrease in cdk2 activity. Fibroblasts deficient in p21 were not susceptible to a reduction in cdk2 activity or growth inhibition by gax overexpression. Localized delivery of the virus to denuded rat carotid arteries significantly reduced neointima formation and luminal narrowing. These data indicate that gax overexpression can inhibit cell proliferation in a p21 dependent manner and can modulate injury-induced changes in vessel wall morphology that result from excessive cellular proliferation. PMID- 9224718 TI - Differential regulation of FUS3 MAP kinase by tyrosine-specific phosphatases PTP2/PTP3 and dual-specificity phosphatase MSG5 in Saccharomyces cerevisiae. AB - The Saccharomyces cerevisiae mating pheromone response is mediated by activation of a MAP kinase (Fus3p and Kss1p) signaling pathway. Pheromone stimulation causes cell cycle arrest. Therefore, inactivation of the Fus3p and Kss1p MAP kinases is required during recovery phase for the resumption of cell growth. We have isolated a novel protein tyrosine phosphatase gene, PTP3, as a negative regulator of this pathway. Ptp3p directly dephosphorylates and inactivates Fus3p MAP kinase in vitro. Multicopy PTP3 represses pheromone-induced transcription and promotes recovery. In contrast, disruption of PTP3 in combination with its homolog PTP2 results in constitutive tyrosine phosphorylation, enhanced kinase activity of Fus3p MAP kinase on stimulation, and delayed recovery from the cell cycle arrest. Both tyrosine phosphorylation and kinase activity of Fus3p are further increased by disruption of PTP3 and PTP2 in combination with MSG5, which encodes a dual specificity phosphatase. Cells deleted for all three of the phosphatases (ptp2delta ptp3delta msg5delta) are hypersensitive to pheromone and exhibit a severe defect in recovery from pheromone-induced growth arrest. Our data indicate that Ptp3p is the major phosphatase responsible for tyrosine dephosphorylation of Fus3p to maintain a low basal activity; it also has important roles, along with Msg5p, in inactivation of Fus3p following pheromone stimulation. These data present the first evidence for a coordinated regulation of MAP kinase function through differential actions of protein tyrosine phosphatases and a dual specificity phosphatase. PMID- 9224719 TI - The 30-kD subunit of mammalian cleavage and polyadenylation specificity factor and its yeast homolog are RNA-binding zinc finger proteins. AB - Cleavage and polyadenylation specificity factor (CPSF), a key component of the mammalian RNA 3'-end processing machinery, consists of four subunits of 160, 100, 73, and 30 kD. Here we report the isolation and characterization of a cDNA encoding the 30-kD polypeptide. Antibodies raised against this protein inhibit cleavage and polyadenylation and coimmunoprecipitate the other CPSF subunits. The protein sequence contains five C3H-zinc-finger repeats and a putative RNA-binding zinc knuckle motif at the carboxyl terminus. Consistent with this observation, the in vitro translated 30-kD protein binds RNA polymers with a distinct preference for poly(U). In addition, an essential S. cerevisiae gene, YTH1, was cloned which is 40% identical to CPSF 30K at the protein level. Extracts prepared from a conditional yth1 mutant have normal cleavage activity, but fail to polyadenylate the upstream cleavage product. Efficient polyadenylation activity can be restored by the addition of purified polyadenylation factor I (PF I). We demonstrate that Yth1p is a component of PF I that interacts in vivo and in vitro with Fip1p, a known PF I subunit. PMID- 9224721 TI - Drosophila Jun relays the Jun amino-terminal kinase signal transduction pathway to the Decapentaplegic signal transduction pathway in regulating epithelial cell sheet movement. AB - We have characterized mutations in the Drosophila homolog of the mammalian proto oncogene c-Jun gene (Djun). We demonstrate that DJUN in the embryo is a downstream target of the JNK signal transduction pathway during dorsal closure formation, and that the function of the JNK/DJUN pathway is to control the localized expression of decapentalegic (dpp), a member of the TGF-beta growth factor family. In contrast to previous observations, we find that both in the embryo and during photoreceptor cell determination, DJUN is not regulated by a pathway that involves MAPK. PMID- 9224720 TI - Drosophila Jun kinase regulates expression of decapentaplegic via the ETS-domain protein Aop and the AP-1 transcription factor DJun during dorsal closure. AB - During Drosophila embryogenesis, ectodermal cells of the lateral epithelium stretch in a coordinated fashion to internalize the amnioserosa cells and close the embryo dorsally. This process, dorsal closure, requires two signaling pathways: the Drosophila Jun-amino-terminal kinase (DJNK) pathway and the Dpp pathway. We have identified mutations in DJun and show that DJNK controls dorsal closure by activating DJun and inactivating the ETS repressor Aop/Yan by phosphorylation. DJun and Aop regulate dpp expression in the most dorsal row of cells. Secreted Dpp then instructs more ventrally located cells to stretch. Our results provide a causal link between the DJNK and Dpp pathways during dorsal closure. Interestingly, in vertebrates, transforming growth factor-beta and c-Jun regulate collagenase gene expression during wound healing, a process that also involves the closing of an epithelial sheath. PMID- 9224722 TI - Coupling of Jun amino-terminal kinase and Decapentaplegic signaling pathways in Drosophila morphogenesis. AB - Dorsal closure in Drosophila embryos involves the migration of two lateral epithelia toward the dorsal midline to establish the dorsal ectoderm. Previous work showed that this morphogenetic movement depends on the activities of a Jun amino (N)-terminal kinase kinase (JNKK) encoded by the hemipterous (hep) gene, and of a JNK encoded by basket. Hep is required for cell determination in the leading edge of migrating epithelia, by controlling specific expression of the puckered (puc) gene in these cells. During dorsal closure, decapentaplegic (dpp), a member of the transforming growth factor-beta (TGF-beta) superfamily, is expressed in the row of cells making up the leading edge of the epithelia. Here, we show that the small GTPases Dcdc42, Drac1, and the Hep JNKK control dpp expression in this migratory process. Appropriate dpp and puc expression in the leading edge also depends on the inhibitory function of the puc gene. Further, our data suggest that the leading edge is the source of a JNK autocrine signal, and exclude a role of Dpp as such a ligand. Dorsal closure couples JNK and dpp signaling pathways, a situation that may be conserved in vertebrate development. PMID- 9224723 TI - Jun in Drosophila development: redundant and nonredundant functions and regulation by two MAPK signal transduction pathways. AB - Drosophila Jun is shown to be involved in different signal transduction pathways and developmental decisions. Dorsal closure, a morphogenetic process occurring during Drosophila embryogenesis, is regulated by Hemipterous (Hep) and Basket (Bsk), homologs of JNKK and JNK, respectively. Embryos lacking Jun activity exhibit a dorsal closure phenotype, very similar to that of bsk and hep mutants, indicating that Jun is a target of Hep/Bsk signaling. In eye and wing development Jun participates in a separate signaling pathway that is comprised of Ras, Raf, and the ERK-type kinase Rolled. In contrast to the strict requirement for Jun in dorsal closure, its role in the eye is redundant but can be uncovered by mutations in other signaling components. The redundant function of Jun in eye development may contribute to the precision of photoreceptor differentiation and ommatidial assembly. PMID- 9224724 TI - Brevetoxin-6 (PbTx-6), a nonaromatic marine neurotoxin, is a ligand of the aryl hydrocarbon receptor. AB - Brevetoxins (PbTx) are a family of marine polyether toxins that exert their toxic action by activating voltage-sensitive sodium channels. Two forms of brevetoxin, PbTx-2 and -3, induce hepatic cytochrome P4501A1, measured as ethoxyresorufin O deethylase (EROD) activity, in redfish and striped bass. P4501A1 induction is transcriptionally regulated through the binding of a ligand, typically a planar aromatic compound, to the aryl hydrocarbon receptor (AhR) and subsequent complex formation with the dioxin response element (DRE), an upstream regulatory region of the CYP1A1 gene. To determine if PbTx, a nonaromatic compound, induced EROD by this mechanism, two sets of experiments were performed. Initially, saturation binding assays with PbTx-2, -3, and -6 were carried out to determine if PbTx-2, 3, or -6 was an AhR ligand. Results showed that PbTx-6 inhibited specific binding of dioxin to the AhR, whereas PbTx-2 and -3 had no effect. Subsequently, gel retardation assays showed that PbTx-6 caused a concentration-dependent increase in AhR-DRE complex formation. The most abundant and neurotoxic forms of brevetoxin, PbTx-2 and -3, did not appear to be involved in this process. However, PbTx-6, the epoxide which is a likely biotransformation product, is at least one of the forms of PbTx involved in EROD induction. PMID- 9224725 TI - Stimulation of osteopontin mRNA expression in HL-60 cells is independent of differentiation. AB - 12-O-Tetradecanoylphorbol 13-acetate (TPA) induces HL-60 cells to differentiate along the monocyte/macrophage pathway and stimulates expression of the extracellular adhesion protein osteopontin (OPN). In this study, the mechanism of TPA-mediated OPN mRNA expression and its relationship to differentiation were investigated. The induction of OPN mRNA by TPA was dose dependently inhibited by staurosporine (0.4-10.0 nM) and chelerythrine (0.1-5.0 microM), indicating that OPN expression requires PKC activation. Furthermore, the mitogen-activated protein kinase kinase (MAPKK) inhibitor, PD 098059 (1.0-10.0 microM), inhibited the effect of TPA in a dose-dependent fashion. Cycloheximide (10 microg/ml) ablated the induction of OPN mRNA by TPA. To determine if OPN mRNA expression was associated with a particular differentiational pathway, HL-60 cells were treated with RA, 9-cis-RA, calcitriol, or sodium butyrate. None of these agents stimulated OPN mRNA. Treatment with TPA subsequent to a 120-h pretreatment with retinoic acid (RA), 9-cis-RA, or calcitriol resulted in a potentiation of the induction of OPN mRNA. These results support a role for protein kinase C (PKC) in promoting OPN expression because each of these agents increased PKC levels. An hOPN promoter/reporter construct was responsive to TPA, indicating that this effect is at the level of transcription. Thus, TPA-stimulated transcription of the OPN gene apparently occurs via a PKC/MAPK-dependent mechanism that is independent of that associated with differentiation and is not dependent on the maturational state of these cells. PMID- 9224726 TI - The role and content of endogenous insulin-like growth factor-binding proteins in bovine articular cartilage. AB - Previous work identified insulin-like growth factor (IGF)-binding proteins (IGF BPs) in chondrocyte culture fluids, but the relationship of these proteins to the composition of intact cartilage was not established. The aim of this work was to analyze the IGF-BP system resident in bovine articular cartilage and to examine its role in IGF-1-regulated proteoglycan (PG) metabolism. Protein extracts of freshly dissected or cultured cartilage slices were analyzed by 125I-IGF-2 ligand blotting. Fresh tissue and basal cultured samples (serum-free) from nine animals, aged fetal to adult, contained two major IGF-BPs of approximately 31,000 and 24,000-21,500 Mr, with the latter doublet being dominant. The 31,000 Mr protein was identified as IGF-BP-2 by specific immunoreactivity with two polyclonal antibodies, and the 24,000-21,500 Mr doublet was identified as IGF-BP-6 by reactivity with a specific polyclonal antibody and by marked preferential affinity for IGF-2 over IGF-1 by ligand blotting. Treatment of the cartilage cultures with 10 ng/ml transforming growth factor-beta (TGF-beta1) for 1 week led to an accumulation of IGF-BP-2, while IGF-BP-6 was unchanged (ligand blots, n = 6 animals). IGF-1 had a similar but much less pronounced effect. The IGF-BP increase following TGF-beta treatment, quantified by charcoal assay, was on average 6-fold, while total protein increased only 1.2-fold (n = 4). By contrast, IGF-1 (10 ng/ml) increased IGF-BP by <2-fold (n = 4), and retinoic acid, at 1 x 10(-8) M was not effective (n = 3). As before, 10 ng/ml TGF-beta or IGF-1 increased proteoglycan synthesis and maintained its homeostasis. IGF-1 analogs with reduced affinity for the IGF-BPs were tested. The effect of an A-chain analog (Thr49, Ser50, Ile51) on PG synthesis was comparable to that of IGF-1, even though the analog had one-half of the IGF-1 affinity for the type I IGF receptor. A B-chain analog, with one-third the receptor affinity of IGF-1, promoted an average 2-fold higher PG synthesis in the linear response range to concentration. Thus, in relation to their respective affinities for the IGF-type I receptor, both IGF analogs were more effective than native IGF-1. These results suggest that an overall effect of the endogenous IGF-BP activity in articular cartilage under the test conditions is the inhibition of IGF-1-stimulated proteoglycan synthesis. PMID- 9224727 TI - The human immunodeficiency virus type 1 Tat protein potentiates zidovudine induced cellular toxicity in transgenic mice. AB - 3'-Azido-2',3'-dideoxythymidine (AZT, zidovudine) is the principal antiretroviral agent in the treatment of AIDS. Although beneficial, AZT remains restricted for human usage because of its severe toxic effects. We examined the AZT sensitivity in transgenic mice expressing HIV-1 one-exon-encoded 72 amino acid Tat (Tat72) and full-length 86 amino acid Tat (Tat86) proteins. Administration of AZT (1 mg/ml) in drinking water for 1 week resulted in a three- to fourfold decrease in hematopoietic progenitors from bone marrow in Tat mice compared to AZT-treated nontransgenic controls as determined by erythroid and granulocyte/macrophage colony-forming unit assays. In liver and thymus, two of the tissues examined, AZT treatment of Tat mice resulted in as much as 80-90% suppression of Mn-superoxide dismutase (Mn-SOD) activity. Other parameters associated with loss of Mn-SOD such as increase in carbonyl proteins and decrease of sulfhydryl content were also significantly enhanced by AZT in Tat mice. Our in vivo study suggests that AZT therapy is associated with oxidative damage affecting cellular functions in several tissues and that Tat is one of the contributory factors in AZT-induced toxicities. The findings of AZT-induced oxidative damage may help to improve the therapeutic index of AZT and other related drugs in AIDS patients. PMID- 9224728 TI - Inhibition by 1alpha,25-dihydroxyvitamin D3 of activin A-induced differentiation of murine erythroleukemic F5-5 cells. AB - 1alpha,25-Dihydroxyvitamin D3 (1alpha,25-(OH)2D3) and other vitamin D3 (VD3) analogs enhanced the inhibitory effect of Activin A on murine erythroleukemia (MEL) cell proliferation and differentiation in a dose-dependent manner. 1alpha,25-(OH)2D3 inhibited differentiation more potently than proliferation by one order of magnitude. The VD3 analog study demonstrated either effect of VD3 on MEL cells via vitamin D receptor (VDR), as evidenced from the close relationship with the reported affinities for VDR. The effects of 1alpha,25-(OH)2D3 were preceded by the suppression of ornithine decarboxylase (ODC) activity, a rate limiting enzyme in polyamine metabolism. Difluoromethylornithine (DFMO), an inhibitor of ODC, inhibited MEL cell proliferation, which was reversed by the simultaneous addition of putrescine, a product of ODC, but did not affect differentiation. 1alpha,25-(OH)2D3 inhibited cell differentiation during the phenotype-expression stage as reflected by the inhibition of beta-globin gene expression, while it inhibited proliferation in the commitment stage. Furthermore, it seems unlikely that the different effects of VD3 on proliferation and differentiation may be a result of upregulation of VDR or nongenomic action. In summary, it was suggested that 1alpha,25-(OH)2D3 inhibited Activin A-induced MEL cell proliferation and differentiation by distinct mechanisms and inhibited the proliferation by inhibiting ODC activity. We demonstrated the presence of 1alpha,25-(OH)2D3 action on leukemic cells at physiological concentration, which was distinct from the pharmacological effect of VD3 reported thus far. PMID- 9224729 TI - Sequence determination and molecular characterization of gigantin, a cytotoxic protein produced by the mould Aspergillus giganteus IFO 5818. AB - Gigantin is a 17-kDa ribonuclease secreted by Aspergillus giganteus IFO 5818. The sequence of the genomic DNA coding for this protein is reported. The deduced amino acid sequence reveals nine amino acid variations with respect to alpha sarcin, a well-characterized ribosome-inactivating protein from A. giganteus MDH 18894. The peptides obtained after tryptic digestion of reduced and carboxyamidomethylated gigantin have been chromatographically separated. The analysis of these peptides in comparison to those originating from alpha-sarcin corroborates the above sequence differences. These do not sensibly modify the conformation of the protein, based on the coincidence of the circular dichroism and fluorescence emission spectra of the two proteins. The obtained results are discussed in terms of the involvement of the distinctive residues in the immunological and catalytic properties that distinguish gigantin from alpha sarcin. PMID- 9224730 TI - Csk phosphorylation and inactivation in vitro by the cAMP-dependent protein kinase. AB - Csk is a protein tyrosine kinase that phosphorylates other protein tyrosine kinases of the Src family and down-regulates their activities. It is not known how Csk is regulated. We investigated the possibility that Csk is regulated through phosphorylation by examining if Csk can serve as an in vitro substrate for a panel of protein kinases. We found that Csk was phosphorylated by the cAMP dependent protein kinase (PKA), but not by protein kinase C, Src, or the fibroblast growth factor receptor kinase. Csk phosphorylation in vitro by PKA is on a serine residue(s) and can reach a stoichiometry of approximately 0.6 mol phosphate per mole of enzyme. Furthermore, incubation with PKA in the presence of ATP and magnesium ion results in a time-dependent decrease in Csk kinase activity. A six-fold decrease in Csk activity (expressed as Vmax/Km ratio) was achieved due to a threefold increase in its Km and a twofold decrease in its Vmax value within 1 h of incubation with the catalytic subunit of PKA and ATP-Mg. Both phosphorylation and inactivation by PKA were blocked by a PKA-specific inhibitor. Csk mutants with a deleted SH2 or SH3 domain retained their ability to be phosphorylated and inactivated by PKA, indicating that the phosphorylation site is located within the catalytic domain. These studies suggest that the cAMP dependent protein kinase can regulate Csk activity. PMID- 9224731 TI - Transport of S-adenosylmethionine in isolated rat liver mitochondria. AB - Mitochondria do not have the enzyme, methionine adenosyltransferase (ATP: L methionine S-adenosyltransferase, EC 2.5.1.6), necessary for the biosynthesis of S-adenosylmethionine. Nevertheless, about 30% of total hepatic S adenosylmethionine resides in the mitochondria and radiolabeled S adenosylmethionine may be isolated from the mitochondria after administration of radiolabeled methionine. This leads to the hypothesis that a carrier-mediated system is responsible for S-adenosylmethionine transport from the cytosol into the mitochondria. We have characterized such a system in isolated rat liver mitochondria. Uptake of S-adenosylmethionine consisted of two components. One component was incorporation of the methyl group into phospholipids as shown by thin-layer chromatography. The second component represented uptake into the mitochondria since addition of excess unlabeled S-adenosylmethionine resulted in efflux of labeled substrate. This countertransport is characteristic of a carrier mediated transport system. Uptake (corrected for incorporation into phospholipids) was saturable with an apparent Km = 8.9 microM and Vmax = 54.3 pmol x mg protein(-1) x min(-1). Uptake was not inhibited by methionine, adenosine, 5'-methylthioadenosine, carnitine, choline, betaine, quinine, or hemicholinium-3. Uptake was inhibited by sinefungin and by S-adenosylhomocysteine (Ki = 53.4 microM). Uptake of S-adenosylmethionine was not dependent on the electrical potential across the mitochondrial membrane. These results indicate that S-adenosylmethionine is taken up into mitochondria via a specific, carrier mediated system. PMID- 9224732 TI - Structure of the gene for type I hexokinase from rat. AB - Based on presumed analogy with the previously characterized gene encoding the Type II isozyme of rat hexokinase (Printz, R.L., Koch, S., Potter, L.R., O'Dougherty, R.M., Tiesinga, J.J., Moritz, S., and Granner, D. K., J. Biol. Chem. 268, 5209-5219, 1993), the locations of splice sites in the gene encoding the rat Type I isozyme of hexokinase have been determined by PCR amplification of intronic DNA. Sequences at the splice sites conform to the consensus sequence, with GT and AG being found at 5' and 3' ends of the introns, respectively. Sizes of exons 1 and 2 were determined directly while others were estimated based on identified splice sites and the previously determined cDNA sequence. These exon sizes were confirmed by PCR amplification, which gave products having sizes consistent with those of introns and exons predicted to be within the amplified sequence. Thus, it is unlikely that the gene encoding the Type I isozyme contains any introns not having analogs in the gene for Type II hexokinase. The deduced structure for the rat Type I hexokinase gene is therefore identical to that for the rat Type II isozyme, and spans over 51 kb. Six tandem repeat sequences of (AC/GT)n have been identified in the 5' flanking region and in introns 10, 11, 12, and 16; this is an unusually high frequency of tandem repeat sequences. PMID- 9224733 TI - Establishment of a novel host, high-red yeast that stably expresses hamster NADPH cytochrome P450 oxidoreductase: usefulness for examination of the function of mammalian cytochrome P450. AB - A novel strain of Saccharomyces cerevisiae useful for expression studies of mammalian microsomal cytochrome P450s was established and named High-red yeast. Hamster NADPH-cytochrome P450 oxidoreductase (P450 reductase) cDNA to be introduced into yeast was isolated from a hamster liver cDNA library. The cDNA was 2421 bp long and contained an entire coding region for 667 amino acids. The NH2-terminal amino acid sequence deduced from the hamster P450 reductase cDNA was identical with that of the enzyme purified from hamster livers except for deletion of the initial methionine. A delta-sequence derived from yeast retrotransposon Ty was cloned and used as a sequence for homologous recombination in a yeast genome. S. cerevisiae YPH500 was transformed with a multi-integration cassette containing the expression unit of the hamster P450 reductase and the delta-sequence. The transformant showing the highest activity of the P450 reductase was named High-red yeast. High-red yeast carried more than six copies of the multi-integration cassettes in a single chromosome and retained the multi integration cassettes over a period of 100 generations under nonselective culture conditions, indicating that this yeast was a mitotically stable transformant. The microsomes prepared from High-red yeast had 20 times the P450 reductase activity of the microsomes prepared from the parental yeast. Due to the high activity of the hamster P450 reductase, the 7-ethoxycoumarin deethylase activity of mouse CYP1A1 expressed in High-red yeast was 250 times higher than the activity of mouse CYP1A1 expressed in the parental yeast. PMID- 9224734 TI - A potent superoxide dismutase mimic: manganese beta-octabromo-meso-tetrakis-(N methylpyridinium-4-yl) porphyrin. AB - Variously modified metalloporphyrins offer a promising route to stable and active mimics of superoxide dismutase (SOD). Here we explore bromination on the pyrroles as a means of increasing the redox potentials and the catalytic activities of the copper and manganese complexes of a cationic porphyrin. Mn(II) and Cu(II) octabrominated 5,10,15,20-tetrakis-(N-methylpyridinium-4-yl) porphyrin, Mn(II)OBTMPyP4+, and Cu(II)OBTMPyP4+ were prepared and characterized. The rate constants for the porphyrin-catalyzed dismutation of O2.- as determined from the inhibition of the cytochrome c reduction are k(cat) = 2.2 x 10(8) and 2.9 x 10(6) M(-1) s(-1), i.e., IC50 was calculated to be 12 nM and 0.88 microM, respectively. The metal-centered half-wave potential was E(1/2) = +0.48 V vs NHE for the manganese compound. Cu(II)OBTMPyP4+ proved to be extremely stable, while its Mn(II) analog has a moderate stability, log K = 8.08. Nevertheless, slow manganese dissociation from Mn(II)OBTMPyP4+ enabled the complex to persist and exhibit catalytic activity even at the nanomolar concentration level and at biological pH. The corresponding Mn(III)OBTMPyP5+ complex exhibited significantly increased stability, i.e., demetallation was not detected in the presence of a 400-fold molar excess of EDTA at micromolar porphyrin concentration and at pH 7.8. The beta-substituted manganese porphyrin facilitated the growth of a SOD deficient strain of Escherichia coli when present at 0.05 microM but was toxic at 1.0 microM. The synthetic approach used in the case of manganese and copper compounds offers numerous possibilities whereby the interplay of the type and of the number of beta substituents on the porphyrin ring would hopefully lead to porphyrin compounds of increased stability, catalytic activity, and decreased toxicity. PMID- 9224735 TI - Analysis of homo- and heterodimerization of retinoid receptors in solution. AB - To characterize the dimerization of retinoid receptors in solution, RAR alpha homodimers and RAR alpha-RXR alpha heterodimers, formed in the absence or the presence of a naturally occurring RA response element (betaRARE) under different ionic conditions, were analyzed by size-exclusion fast protein liquid chromatography and sucrose density gradient sedimentation. In the presence of [3H]RA both RAR alpha and RXR alpha existed primarily as monomers of 50 kDa in solutions containing 80 mM KCl. However, when betaRARE was included in these incubations, a 40-fold increase in the occurrence of both the RAR alpha homodimers and the RAR alpha-RXR alpha heterodimers (125 kDa) was observed. The presence of RAR alpha and RXR alpha in the betaRARE-associated homo- and heterodimers was confirmed by the positive interaction of the receptors with the specific antibodies. Both RAR alpha homodimers and RAR alpha-RXR alpha heterodimers bound betaRARE even in the absence of the ligand RA with the heterodimer showing a 2- to 4-fold greater affinity than the homodimer for the DNA binding element. When the receptors were incubated in solutions of increasing ionic concentration (50-300 mM KCl), a decrease in the amount of both RAR alpha homodimers and RAR alpha-RXR alpha heterodimers was accompanied by a corresponding increase in the monomeric fraction even in the presence of betaRARE, suggesting that the high salt concentrations inhibit the surface to surface interactions between the monomers. These observations suggest that in vivo, as in solution, the formation of a stable retinoid receptor dimer complex is dependent upon both receptor-receptor and receptor-RARE interactions. PMID- 9224736 TI - Ascorbate and alpha-tocopherol prevent apoptosis induced by serum removal independent of Bcl-2. AB - Cells require serum to maintain growth in vitro. Serum provides growth and survival factors and its removal causes an oxidative stress that induces peroxidations in membrane lipids and development of programmed cell death (apoptosis) in some cells. Cells containing Bcl-2 are partially protected against both lipid peroxidation and apoptosis and some cell lines, such as Daudi, which lack this protein, are very sensitive to serum removal. Thus, cells are grown for 48 h in the absence of fetal calf serum and apoptotic cells are scored. HL-60 cells containing a moderate amount of Bcl-2 show 30% apoptosis, while 55% cells are apoptotic of the Bcl-2-negative Daudi cell population. Apoptosis is reduced to 15% in the transiently transfected Daudi/Bcl-2 cells. Ascorbate (Asc) and alpha-tocopherol (alphaTOH) can prevent lipid peroxidation and apoptosis caused by serum withdrawal, when added to culture media, even in the absence of Bcl-2. Also, these two antioxidants increase survival of cells grown in the absence of serum independent of their Bcl-2 content. Immunostaining and quantification of Bcl-2 show that HL-60 cell line is a heterogeneous population relative to the expression of Bcl-2. When these cells are grown in the presence of serum, cells lacking Bcl-2 survive, but no Bcl-2-negative cells survive without serum. Part of this population of Bcl-2-negative cells is rescued by Asc and alphaTOH. Antioxidants effective at the plasma membrane such as Asc and alphaTOH can protect cells from oxidative damage and prevent apoptosis independent of Bcl-2 content. PMID- 9224738 TI - Oxidative modification and nitration of human low-density lipoproteins by the reaction of hypochlorous acid with nitrite. AB - Hypochlorous acid (HOCl) reacts with nitrite (NO2-) at a molar ratio of 1:1 yielding an equimolar amount of nitrate. The rate of this reaction follows the dissociation of hypochlorous acid and decreases with the increasing of pH from 4 to 10 as assayed by stopped-flow analysis, suggesting that HOCl, not hypochlorite, is the reactant. The second-order rate constant at pH 7.2, 25 degrees C, was estimated as (7.4 +/- 1.3) x 10(3) M(-1) s(-1), a rate considerably higher than that of the Fenton reaction (42 M(-1) s(-1)). In human low-density lipoproteins (LDL) the reaction led to a loss of beta-carotene and alpha-tocopherol. The NO2-/HOCl mixture initiated lipid peroxidation in LDL, whereas NO2- or HOCl alone had only little effect. When LDL was added immediately after mixing of NO2- with HOCl, no loss of antioxidants or accumulation of lipid peroxidation products was observed, suggesting that a short-lived reactive intermediate, previously postulated as nitryl chloride, is the reactive species. The mixture NO2-/HOCl as well as peroxynitrite led to the formation of 3 nitrotyrosine in LDL as assayed using a monoclonal anti-nitrotyrosine antibody. Furthermore, incubation of J774.2 macrophage-like cells with LDL, pretreated with the NO2-/HOCl mixture, led to increased cellular accumulation of cholesterol. Thus modification of LDL caused by the reaction of nitrite with HOCl contributes to the formation of cholesterol-rich cells, a key feature of the early atherosclerotic lesion. PMID- 9224737 TI - Characterization of human liver inducible nitric oxide synthase expressed in Escherichia coli. AB - We have cloned the human liver inducible isoform of nitric oxide synthase (NOS) into an Escherichia coli expression vector and have expressed and purified the enzyme. The protein has been expressed with and without a polyhistidine tail. In both cases, expression of functional protein requires coexpression with calmodulin and inclusion of tetrahydrobiopterin (H4B) in the purification buffers. Unlike the constitutive isoforms of NOS, this isoform is unstable in the absence of L-arginine (L-Arg) and H4B toward loss of the heme group and the formation of a low-spin species spectroscopically distinct from that of the cofactor-bound protein. The enzyme purified in the presence of both L-Arg and H4B is highly active, with a Vmax of approximately 800 nmol NO min(-1) mg(-1) and a Km for L-Arg of 22 microM. The cytochrome c reductase activity is 38,000 nmol x min(-1) mg(-1). Similar values are obtained for the enzyme with and without the polyhistidine tail. Ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid does not inhibit the activity of the protein, nor is the activity of the enzyme increased by the addition of exogenous calmodulin and/or Ca2+. These findings contrast with an earlier report, based on experiments with extracts of COS-1 cells expressing the recombinant enzyme, that the enzyme responds to changes in the Ca2+ concentration. The human hepatic isoform is similar in its properties to the inducible NOS isoform purified from macrophages. PMID- 9224740 TI - Is clonality equivalent to malignancy: specifically, is immunoglobulin gene rearrangement diagnostic of malignant lymphoma? PMID- 9224739 TI - Phosphoenolpyruvate carboxylase protein kinase from soybean root nodules: partial purification, characterization, and up/down-regulation by photosynthate supply from the shoots. AB - Phosphoenolpyruvate carboxylase (PEPC) kinase was partially purified about 3000 fold from soybean root nodules by a fast-protein liquid chromatography protocol. This protein-serine kinase has an apparent native molecular mass of about 30,000 as estimated by size-exclusion chromatography. Following electrophoresis of this partially purified PEPC-kinase preparation in a denaturing gel containing dephospho maize leaf PEPC as substrate, the in situ renaturation and assay of protein kinase activity revealed two, PEPC-dependent kinase polypeptides with molecular masses of about 32 and 37 kDa. The approximately 32-kDa polypeptide was significantly more active than the approximately 37-kDa catalytic subunit. The activity of this partially purified PEPC kinase, and a less purified sample, was Ca2+-insensitive. This protein kinase preparation was able to phosphorylate purified PEPCs from soybean nodules, maize leaves, and a sorghum recombinant C4 PEPC. In contrast, this PEPC kinase was unable to phosphorylate a phosphorylation site mutant form of sorghum C4 PEPC (S8Y), two other soybean nodule phosphoproteins [nodulin-26 and nodulin-100 (sucrose synthase)], bovine serum albumin, and histone III-S. Following in vitro phosphorylation of purified dephospho soybean nodule PEPC from stem-girdled plants by the partially purified nodule PEPC kinase, the former's activity and sensitivity to L-malate inhibition increased and decreased, respectively. Notably, the Ca2+-independent PEPC kinase activity in nodules from illuminated plants was markedly greater than that in nodules harvested from plants subjected to stem girdling or prolonged darkness. Furthermore, the kinase activity in nodules was controlled reversibly by illumination and extended darkness pretreatments of the parent plants, suggesting that photosynthate supply from the shoots is likely responsible for these striking changes in PEPC kinase activity observed in planta in the legume nodule. PMID- 9224741 TI - Human parvovirus B19 in bone marrows from adults with acquired immunodeficiency syndrome: a comparative study using in situ hybridization and immunohistochemistry. AB - Human parvovirus B19, which infects and lyses erythroid precursors, can cause severe anemia in patients with immunodeficiency. The incidence of parvovirus infection in adult acquired immunodeficiency syndrome (AIDS) patients is unknown. Eighty-one archival formalin-fixed, paraffin-embedded (FFPE) bone marrow biopsies from 73 AIDS adults were immunostained with monoclonal R92F6 against B19 VP1 and VP2 capsid proteins using streptavidin peroxidase and streptavidin alkaline phosphatase techniques. In addition, the same tissues were hybridized in situ with a digoxigenin-labeled parvovirus B19 DNA probe. Five FFPE bone marrows, from 3 HIV-negative patients with positive immunoglobulin M (IgM) serology for parvovirus B19, and 1 parvovirus B19-infected fetal liver were positive controls. By immunoperoxidase, all tissues were negative with R92F6 except the fetal liver, which exhibited strong positivity predominantly in viral inclusions. With immunoalkaline phosphatase, all positive controls were immunoreactive with R92F6; however, the AIDS marrows were negative. With in situ hybridization (ISH), all positive controls and 7 of 81 (8.6%) of AIDS marrows were positive for B19 parvovirus DNA. We conclude that ISH is more sensitive than R92F6 immunohistochemistry in parvovirus B19 detection. A small but significant number of bone marrows from AIDS adults shows evidence of human parvovirus B19 infection. PMID- 9224742 TI - EWS/FLI-1 fusion transcripts in three peripheral primitive neuroectodermal tumors of the kidney. AB - Although primitive neuroectodermal tumor (PNET) is a well-recognized entity, its renal localization as a primary site has not been appreciated. Only nine cases of renal PNET exist in the literature. The paucity of renal PNET could be explained by the lack of objective diagnostic techniques that would facilitate its distinction from other primitive round cell tumors of the kidney, such as the more widely recognized monophasic Wilms' tumor and clear-cell sarcoma of the kidney (CCSK), as well as renal carcinoid, or neuroblastoma invading the kidney from the adjacent adrenal gland. The recently identified specific fusion transcripts detectable by reverse transcription polymerase chain reaction (RT PCR) have provided us with a valuable tool for the detection of renal PNET. This article reports three renal PNET that expressed EWS/FLI-1 fusion transcripts by RT-PCR, in addition to positive staining for MIC2 protein and neuron-specific enolase (NSE). One also exhibited dense core granules in cell processes by electron microscopy. Employment of such methodology will lead to a more accurate classification of renal tumors. PMID- 9224743 TI - Role of CD44 in nonpalpable T1a and T1b breast cancer. AB - Primary infiltrating ductal carcinomas (IDCs) of the breast which measure less than 0.5 cm (T1a lesions) and between 0.5 and 1.0 cm (T1b lesions) are associated with a small risk of nodal metastasis. The role of axillary dissection in T1a and T1b breast cancer is controversial. In the absence of axillary dissection, comparable prognostic information might be obtained by examination of the primary cancer. The adhesion molecule CD44 represents a family of transmembrane proteins that mediate cell-cell and cell-matrix interactions. Previous investigators have correlated expression of CD44 and its isoforms with prognosis in breast cancer. We investigated the value of CD44 isoform expression as a predictor of nodal metastases in nonpalpable T1a and T1b IDC. Monoclonal antibody against the standard form of CD44 (CD44s) and polyclonal antibody directed against the variant isoform (CD44v6) was tested on 34 cases of nonpalpable node-negative infiltrating ductal carcinoma (IDC) less than 1.0 cm and 9 cases of nonpalpable node-positive IDC less than 1.0 cm. The expression of CD44s was significantly decreased in node-positive T1a and T1b IDC versus node-negative T1a and T1b IDC (11% vs 65%). In contrast, 97% of the node-negative IDC and 100% of the node positive IDC expressed the CD44v6 isoform. We conclude that CD44s expression is significantly altered in T1a and T1b IDC with nodal metastases but that the CD44v6 isoform does not correlate with nodal metastases in nonpalpable stage T1a and T1b IDC. PMID- 9224744 TI - Loss of heterozygosity of chromosome 14q in low- and high-grade meningiomas. AB - Abnormalities of chromosome 22q have been well studied as a major molecular genetic event in meningiomas. Chromosome 14q loss has also been shown to be a common phenomenon. However, only a few studies have reported molecular genetic changes of this chromosome in meningioma. In this study, we examined 41 sporadic meningiomas of different histological subtypes and grades with 15 polymorphic microsatellite markers covering a wide region on chromosome 14q. Overall, 37% (15 of 41) of cases showed loss of heterozygosity for one or more allelic markers. Thirty percent (10 of 33) of benign tumors showed allelic losses, whereas 62.5% (5 of 8) of high-grade meningiomas showed loss of heterozygosity. There were altogether eight cases of partial deletions and seven cases of probable monosomy of 14q. Allelic losses of 14q were also commonly seen in recurrent tumors (3 of 3, 100%), parasagittal tumors (4 of 7, 57%), and tumors with transitional subtype (7 of 14, 50%). Among the tumors with allelic losses of 14q, all except one concurrently showed loss of heterozygosity for markers on 22q by our previous study. Of the eight cases with partial deletions, one showed losses on 14q11.1 31, two showed deletions on 14q24.3-31, three showed losses at 14q32.1-32.2, and the remaining two showed deletions at 14q24.3-32.2. We therefore defined two cluster regions of deletion on chromosome 14q: 14q24.3-31 and 14q32.1-32.2. Our studies suggested that more than one tumor suppressor gene(s) residing on distinct regions of chromosome 14q are important in the development and atypical or anaplastic changes in meningiomas. PMID- 9224746 TI - Peribiliary vascular plexus in primary sclerosing cholangitis and primary biliary cirrhosis. AB - The peribiliary vascular plexus plays an important role in physiology of bile flow. Disturbance of the microcirculation may contribute to ductal injury, but little is known about alterations in the vascular supply of small bile ducts in liver disease. Immunoperoxidase stains for vascular endothelium (Ulex europaeus, factor VIII-related antigen, CD34) were used to study the peribiliary vascular plexus in 20 cases of primary sclerosing cholangitis (PSC) and 27 cases of primary biliary cirrhosis (PBC), two diseases characterized by bile duct destruction. Normal liver from 10 autopsy cases of sudden cardiac death was used as a control. Interlobular bile ducts (20- to 80-microm diameter) were identified on AE1/AE3 immunostain; vessels adjacent to the basement membrane of these ducts were counted. Normal interlobular bile ducts had an average of 2.15 vessels per duct (range, 1.68 to 2.71). Few PBC or PSC cases had a normal number of peribiliary vessels. There was a trend toward vasopenia at higher stage, although vascular loss was noted in early stages as well. The pattern of vascular loss was different for the two diseases; in PSC, the periductal capillaries were often preserved but were pushed away from the basement membrane by concentric deposits of collagen. Small residual vessels could be identified within fibrous scars of obliterated bile ducts in PSC. In 4 stage 3 or 4 PSC cases with little bile duct injury, vessel/duct ratio approached normal levels. In PBC, vessels were obliterated in areas of granulomatous inflammation and heavy lymphocytic infiltrate around bile ducts. In conclusion, loss of peribiliary vessels is common in PSC and PBC. Vessel loss is seen in early stages and may contribute an element of ischemia to continued small bile duct loss but is probably secondary to the inflammatory process. PMID- 9224745 TI - p53 expression and proliferative activity in Bowen's disease with or without chronic arsenic exposure. AB - A comparative study of Bowen's disease (BD) with or without chronic arsenic exposure may contribute to understanding arsenic carcinogenesis. We compared the p53 overexpression and proliferative activity of 26 cases of BD with chronic arsenic exposure (group I) and 22 comparable cases of BD without chronic arsenic exposure (group II) by immunohistochemical method on formalin-fixed, paraffin embedded tissues with antibodies PAb1801 and MIB-1, respectively. We also included in this study two squamous cell carcinomas that developed from BD in group I and one in group II. Two paired BD lesions in the same individual of one patient in group I and of three patients in group II were also studied. The significant p53(+) (>10% stained cells) rates were 42.3% (11 of 26) and 9.1% (2 of 22) for groups I and II, respectively, and the difference was statistically significant (P = .01). The p53 expression in different lesions of the same individual remained consistently the same. Squamous cell carcinomas that developed in 2 cases of p53(+) BD in group I were also positive, but the one in 1 case of p53(-) BD in group II was negative. No significant statistical difference in proliferative activity was found between group I BD and group II BD (P= .769), nor between p53(+) cases (>10% stained cells) and p53(-) cases (<10% stained cells) in group I BD (P = .519). This study showed that significant overexpression of p53 protein was higher in BD with chronic arsenic exposure. Therefore, arsenic carcinogenesis of BD might be different from that of BD unrelated to arsenic, and alteration of p53 plays a more important role in the pathogenesis of BD with chronic arsenic exposure. Overexpression of p53 was not a prerequisite for developing squamous cell carcinoma and was not affected by proliferative activity. PMID- 9224747 TI - Presence of mycobacterial DNA in sarcoidosis. AB - In 11 of 35 clinically proven cases of sarcoidosis, we detected DNA sequences coding for the mycobacterial 65-kDa antigen. In four cases, the sequences were homologous to Mycobacterium avium; seven sequences were related to other nontuberculous Mycobacteria. The insertion sequence 1110, characteristic for Mycobacterium avium, was present in three cases. The insertion sequence 6110 of the Mycobacterium tuberculosis complex (M tuberculosis, africanum, bovis, BCG) was not detectable in any of the 11 cases, ruling out the presence of members of the Mycobacterium tuberculosis complex. Therefore, it seems reasonable to speculate about a mycobacterial cause in some cases of sarcoidosis. PMID- 9224748 TI - Human herpesvirus-8-associated body cavity-based lymphoma in human immunodeficiency virus-infected patients: a unique B-cell neoplasm. AB - Human immunodeficiency virus (HIV)-related body cavity-based lymphomas (BCBLs) are known to exhibit unusual clinical, immunophenotypic, and genotypic features, and have recently been found to harbor DNA sequences of a new human herpesvirus, designated Kaposi's sarcoma-associated herpesvirus (KSHV) or human herpesvirus 8 (HHV-8). The authors have encountered eight cases of HHV-8-associated BCBL in HIV infected patients. A literature search revealed an additional 50 reported cases of HIV-related BCBL, as well as reports of several other disorders associated with HHV-8 DNA. Comprehensive analysis of the clinical and pathobiological features of all 58 known cases of HIV-related BCBL shows it to be a unique B-cell neoplasm with a strong propensity for body-cavity involvement without mass lesions and with little or no dissemination, poor prognosis, high grade usually immunoblastic morphology, late B-cell phenotype and genotype, no associated c-myc gene rearrangement, frequent presence of Epstein-Barr virus (EBV) genome, and uniform association with HHV-8 DNA. Considering these features in the context of other disorders associated with HHV-8 DNA, HHV-8 appears to play a causal role in BCBL, possibly in concert with EBV, and may induce this lymphoma through dysregulation of cytokines, particularly interleukin-6, or infection of an unusual B-cell subset. The characteristics of HHV-8-associated BCBL suggest a possible role for antiherpes or anticytokine agents in the treatment of this lymphoma. PMID- 9224749 TI - Expression of CD44 standard form and variant isoforms in non-small cell lung carcinomas. AB - CD44, a cell adhesion molecule, has been implicated in tumor invasion and metastasis in certain malignancies. We studied the expression of CD44 standard (CD44s) and variant isoforms (CD44v) in 98 non-small cell lung carcinomas (NSCLCs) by immunohistochemistry and correlated the observations with clinical outcome. Formalin-fixed, paraffin-embedded archival tissues from 49 squamous cell carcinomas (SCCs) and 49 adenocarcinomas (ACs) were immunostained after microwave irradiation with monoclonal antibodies against CD44s and CD44v3, v4/5, v6, v7/8, and v10, and the results were correlated with histological tumor type, tumor stage, recurrence, and survival rates. SCCs of the lung showed strong membranous expression of each of the CD44s, v3, v4/5, v6, and v10 proteins in comparison with ACs (P < .0001). Staining for CD44 v4/5 was overwhelmingly positive in SCCs (72%) as compared with ACs (2.2%). Intense immunoreactivity for CD44v6 was present in 19 of 20 (95%) metastatic lung carcinomas. The bronchiolar basal cells and alveolar pneumocytes were positive for CD44s, v3, and v6. CD44s and variant isoform expression did not correlate with tumor stage, recurrence, and survival rates. In conclusion, there is significant immunopositivity of CD44s and variant isoforms in SCCs over ACs of the lung. Expression of CD44v6 may suggest an increased risk for local lymph node metastasis in NSCLCs. CD44v4/5 reactivity may be useful to discriminate squamoid differentiation in poorly differentiated NSCLCs. PMID- 9224750 TI - Giant cell tumor of tendon sheath is a polyclonal cellular proliferation. AB - Giant cell tumor of tendon sheath (GCTTS) is a common soft tissue tumor. Immunophenotypical evidence suggests it is of synovial cell origin. There is controversy regarding the underlying nature of this lesion, specifically whether it is a neoplastic or nonneoplastic (ie, reactive or hyperplastic) process. Karyotypic abnormalities have been identified in GCTTS and interpreted as evidence of neoplasia, although the finding of similar karyotypic abnormalities in unequivocally nonneoplastic proliferations raises questions about using such findings to define a neoplasm. In an attempt to resolve this uncertainty, a polymerase chain reaction (PCR)-based assay for methylation of the X-linked human androgen receptor gene (HUMARA) was used to assess whether GCTTS is a clonal or polyclonal proliferation. DNA was isolated from formalin-fixed, paraffin-embedded tissue blocks from eight cases of digital GCTTS in female subjects; two cases of hepatocellular carcinoma (HCC) were used as clonal controls. Seven of eight cases of GCTTS were informative, and each showed a polyclonal proliferation, whereas both cases of HCC were clonal. Our results indicate that GCTTS is a nonneoplastic proliferation, if one accepts that a population of cells forming a tumorous mass must show clonality to be classified as a neoplasm. Our results emphasize that simple karyotypic abnormalities do not define a neoplasm. It remains to be determined whether GCTTS is a reactive or hyperplastic process. PMID- 9224751 TI - Rapid polymerase chain reaction-based detection of the causative agent of cat scratch disease (Bartonella henselae) in formalin-fixed, paraffin-embedded samples. AB - Bartonella (formerly Rochalimaea) henselae (Bh) plays a central role in cat scratch disease. A polymerase chain reaction (PCR)-based assay that can detect Bh DNA in formalin-fixed, paraffin-embedded (FF-PE) samples would have utility in the evaluation of processed lymph nodes suggestive of this disorder. Fresh or FF PE cultures of Bh and related species were analyzed. Thirteen lymph nodes (12 FF PE and one fresh cell suspension) with necrotizing suppurative granulomatous inflammation and seven FF-PE negative control lymph nodes were also evaluated. PCR was performed using a novel, hemi-nested protocol. Amplified products were analyzed by gel electrophoresis. The fresh and FF-PE Bh cultures showed a specific PCR product with an analytical sensitivity of 0.5 pg bacterial DNA. Seven (54%) of 13 clinical lymph node samples with morphological features suggestive of cat scratch disease also had detectable Bh DNA, whereas none of the seven negative control lymph nodes yielded positive results. We have designed a rapid and sensitive PCR test that can reliably detect Bh DNA in fresh and FF-PE samples. Our findings indicate that this assay has clinical utility in the diagnosis of cat scratch disease. PMID- 9224752 TI - HER-2/neu gene amplification status in prostate cancer by fluorescence in situ hybridization. AB - HER-2/neu expression has been established as a prognostic factor in breast and other cancers. In prostate cancer (PC), a similar predictive role has been hindered by variable immunohistochemical (IHC) results. The authors studied DNA amplification of the HER-2/neu gene on 4-microm sections obtained from 62 formalin-fixed, paraffin-embedded PCs by fluorescence in situ hybridization (FISH). The results were compared with HER-2/neu protein expression as determined by IHC and correlated by logistic regression analysis with Gleason tumor grade, DNA ploidy, serum prostate specific antigen (PSA), and pathological stage. The HER-2/neu gene was localized using the Oncor (Gaithersburg, MD) digoxigenin labeled unique sequence probe. Amplified PCs had at least 20 malignant cells, with 5 or more copies of the sequence. Amplification of HER-2/neu correlated with Gleason score (P = .0001). The mean Gleason score of unamplified tumors was 5.7 and that of amplified tumors was 7.5. Nondiploid tumors had a significantly greater rate of HER-2/neu amplification compared with diploid tumors (P = .0003). Of the 62 cases evaluated by IHC and FISH, 18 cases (29%) were overexpressed by IHC, and 27 cases (44%) were amplified by FISH. A trend for similar HER-2/neu status in each PC by the two methods did not reach statistical significance (P = .23). HER-2/neu amplification by FISH was associated with advanced pathological stage; however, this relationship reached only near-statistical significance (P = .06). There was no correlation of HER-2/neu amplification by FISH with patient age or preoperative serum PSA levels. The authors conclude that HER-2/neu gene amplification status can be determined by FISH on archival prostate cancer specimens, significantly correlates with high tumor grade and nondiploid DNA content, and is more frequently encountered in tumors with advanced pathological stage. Also, FISH is more sensitive than IHC for detection of abnormalities in the HER-2/neu gene, and further studies should be undertaken to determine whether a FISH-based HER-2/neu detection method may prove of importance in the prediction of prognosis and planning of therapy in prostate cancer patients. PMID- 9224753 TI - Extranodal head and neck lymphomas in Guatemala: high frequency of Epstein-Barr virus-associated sinonasal lymphomas. AB - Sinonasal lymphomas of T cell or natural killer cell (T/NK cell) phenotype represent a subset of extranodal head and neck lymphomas. T/NK cell sinonasal lymphomas have been described in diverse geographic settings, including China, Japan, Peru, Northern Europe, and North America. The frequency of these lymphomas is highly dependent on the geographic location in which they occur, their incidence being low in Europe and North America and relatively high in Asian countries and in Peru. Regardless of their geographic location, they are typically associated with the Epstein-Barr virus (EBV). Few studies have addressed the relative frequency of sinonasal lymphoma within the group of extranodal head and neck lymphomas. We investigated the anatomic distribution, immunophenotypical profile, and EBV status of 33 cases of extranodal head and neck lymphoma from patients in Guatemala. The anatomic distribution of these lymphomas is similar to that seen in Asian countries: 17 (52%) in the sinonasal area, five (15%) in the palate, and 11 (33%) in other locations. Fifteen (88%) of the 17 sinonasal lymphomas showed a T or null cell phenotype with a strong association with EBV by in situ hybridization. Most Guatemalan patients with these lymphomas were of Mayan descent. In Guatemala, the relative frequency of sinonasal lymphomas within the group of head and neck lymphomas is significantly higher than that reported for Western countries. In addition, the relative frequency of T/NK versus B cell sinonasal lymphomas is higher than that described in North America and similar to that observed in Asian countries and Peru. PMID- 9224754 TI - Recognition of normal, neoplastic, and fetal airway epithelial cell membranes by two monoclonal antibodies. AB - The reactivity of two rat monoclonal antibodies was studied. These antibodies, A2R and A2C, bind a 32 kDa alveolar type II cell membrane receptor for surfactant protein A. A2R and A2C also bind apical cell membranes of ciliated and nonciliated cells of the conducting airways. Because this reactivity suggested possible utility in targeting those cells for therapeutic gene transfer, the binding activity of these two antibodies was examined in human tissues. In conducting airways, A2R and A2C bound apical epithelial cell membranes throughout the embryologic period studied: from 15 weeks of gestation, through maturity. Reactivity was more restricted to ciliated cells of the airways as maturation progressed. In the peripheral lung, A2C and A2R only bound most cells in the early developing lung, but mainly type II cells in mature lungs. Other normal tissues recognized by these antibodies included crypt lining cells of the adult and fetal stomach, large bile duct epithelium, and pancreatic acinar cells. All of these cells derive from embryonic foregut endoderm. Other normal tissues, both of endodermal and nonendodermal origin, were negative. Pulmonary carcinomas were studied. A2C and A2R recognized all non-small cell carcinomas of the lung tested. In contrast, none of the small cell carcinomas or carcinoid tumors of the lung were recognized by these antibodies. The function of p32 in these diverse cell types is not clear, but whatever its role in these tissues, antibodies versus p32 may potentially be used to target gene or drug therapy to the normal or malignant cells they recognize. PMID- 9224755 TI - Histomorphological patterns of renal amyloidosis: a correlation between histology and chemical type of amyloidosis. AB - A retrospective study was conducted to investigate whether there was a correlation between the histological pattern of renal amyloidosis, the chemical type of amyloid protein involved and the clinical presentation. Eighteen consecutive cases of systemic amyloidosis that had renal biopsies processed and examined histopathologically at the Department of Pathology, Faculty of Medicine, University of Malaya, Kuala Lumpur were reviewed. The age range of patients was 25 to 64 yrs (mean, 46 yrs). The male:female ratio was 2.6:1. Three patients were Malay, 9 Chinese, 3 Indian, 1 Indonesian, 1 Iban, and 1 Bisaya. According to the predominant site of amyloid deposition, 14 cases showed a glomerular pattern and 4 a vascular pattern. 8 cases were designated as 2 anti-human amyloid-A (AA) amyloidosis on the basis of permanganate-sensitivity and immunoreactivity of deposits with anti-human AA protein antibody. Ten cases contained deposits that were permanganate-resistant and nonimmunoreactive for AA protein and were designated as AL in type. The histomorphologic pattern of renal amyloidosis did not provide a reliable means of differentiating AA from AL amyloidosis. The glomerular pattern tended to present with renal manifestations such as nephrotic syndrome and chronic renal failure, whereas the vascular pattern tended to present with nonrenal manifestations such as diarrhoea. These findings may have a bearing on the pathophysiology of amyloidosis and provide clues to appropriate management. PMID- 9224756 TI - Salivary gland lymphoid infiltrates associated with lymphoepithelial lesions: a clinicopathologic, immunophenotypic, and genotypic study. AB - The criteria for distinguishing benign lymphoepithelial lesions (BLEL) from low grade B-cell lymphomas of mucosa-associated lymphoid tissue (MALT) type in salivary glands and the significance of genotypically documented clonality in this setting are controversial. In addition, the clinical implications of a neoplastic diagnosis are unclear. The histopathologic features of 68 specimens from 49 patients with at least one salivary gland biopsy with LEL together with available clinical data were, therefore, reviewed. Paraffin section immunohistochemical (IHC) stains for kappa, lambda, CD3, CD20, and CD43; in situ hybridization (ISH) for kappa and lambda; and polymerase chain reaction (PCR) for immunoglobulin (Ig) HC rearrangement were performed. The 61 salivary gland specimens were classified as BLEL-13, BLEL with monocytoid B-cell (MBC) halos (BLEL-halo-8), low grade B-cell lymphoma of MALT type with confluent zones of MBC or other atypical lymphocytes (ML-MALT-24), low grade B-cell lymphoma of MALT type with monoclonal plasma cells (ML-MALT-PC-12), and high grade B-cell lymphoma of MALT type (MALT-high grade-4). Soft tissue and perineural invasion was not observed in BLEL and was most common in the MALT lymphomas. Lymph node involvement was identified in six patients at the time of their salivary gland MALT lymphomas but in none with BLEL. CD43+ B cells were seen most commonly in ML MALT but were present in all other categories except MALT-high grade. Clonal B cells were identified by PCR in 5 of 12 BLEL, 5 of 8 BLEL-halo, 17 of 22 ML-MALT, 6 of 10 ML-MALT-PC, and 3 of 3 MALT-high grade biopsies. All ML-MALT-PC were clonal by ISH or IHC. Repeat biopsies in 14 patients most commonly showed a BLEL/ML-MALT lesion in an ipsilateral or contralateral salivary gland with one transformation to a MALT-high grade. Although only a few patients are known to have received chemoradiation or radiation therapy, most patients with low-grade lesions have pursued an indolent course. These data show the presence of two types of borderline lesions within the spectrum of lymphoid proliferations associated with salivary gland LEL. One has clonal B cells without histological features of neoplasia and the other nonconfluent MBC extending beyond the confines of LEL ("halos"). They share some features with the infrequent nonneoplastic BLEL and others with the more common low-grade B-cell lymphomas of MALT. A few high-grade B-cell lymphomas of MALT were also identified including a rare example of transformation from a low- to high-grade lesion. The optimal therapeutic approach for the borderline and low-grade lesions and the reason why so many of the lymphoproliferative lesions associated with LEL remain localized to the neck remain to be defined. PMID- 9224757 TI - Loss of H19 imprinting and up-regulation of H19 and SNRPN in a case with malignant mixed Mullerian tumor of the uterus. AB - In several human cancers, it has been recently reported that abnormally altered status of genomic imprinting is related to oncogenesis. In this study, we investigated the expression of three imprinted genes in a case with malignant mixed Mullerian tumor of the uterus (MMMT). In the tumor, expression of H19 showed marked upregulation (6.3-fold) with biallelic expression compared with that in the corresponding normal myometrium. The 5'-promoter region of H19 was hypomethylated in the tumor, whereas it was hemimethylated in the myometrium. Expression of the small nuclear ribonucleoprotein polypeptide N gene (SNRPN) was also upregulated by 1.9-fold. However, the insulin-like growth factor II gene (IGF2) was expressed at low levels in both myometrium and MMMT. The overexpression of H19 is caused by reactivation of the repressed allele of H19 due to demethylation of CpG islands within its 5'-promoter region. Whether upregulation of SNRPN is caused by its biallelic expression remains undetermined because restriction fragment length polymorphisms (RFLP) sites were not informative in SNRPN and IGF2. In conclusion, H19 and SNRPN may play significant roles in the tumorigenesis of MMMT and H19 may have tumor-promoting activity in addition to its known tumor-suppressing activity, probably depending on the tissue and the local milieu. PMID- 9224758 TI - Multifocal hemangioendothelioma of the fetus and placenta. AB - A case of multifocal hemangioendothelioma of the liver, adrenal gland, and placenta is reported. The histological appearance of the tumor is consistent with an infantile hemangioendothelioma, type 2. Multifocal development is the most obvious explanation for the disease but the possibility that this represents malignant placental neoplasm with metastases requires consideration. PMID- 9224759 TI - Dedifferentiated acinic cell carcinoma of the parotid gland: a distinct rarely described entity. AB - A case of dedifferentiated acinic cell carcinoma of the parotid gland is presented. A 46-year-old man presented with a parotid gland mass. At surgery the tumor was found adherent to the temporal bone and cervical adenopathy was present. Treatment included radical parotidectomy and intraoperative radiotherapy. Histologically, the tumor was a composite of a usual low-grade acinic cell carcinoma and high-grade, poorly differentiated carcinoma. Cervical lymph node metastases were composed entirely of high-grade carcinoma. Immunohistochemically, both low- and high-grade malignant components were negative for p53 oncoprotein expression. Moreover, polymerase chain reaction and nonisotopic single-stranded conformational polymorphism analyses were consistent with a germ line configuration of the p53 gene, exons five through eight, in both low- and high-grade elements of the tumor. The literature on this unusual variant of acinic cell carcinoma is reviewed. PMID- 9224760 TI - Mutations in the p53 gene in pulmonary blastomas. PMID- 9224761 TI - Molecular chaperones and the cytoskeleton. AB - Heat shock proteins, first observed because they are preferentially synthesized by organisms exposed to heat or other physiological stress, are also synthesized constitutively. These proteins are divided into several families, namely, HSP100, 90, 70, 60 (chaperonin), and the small heat shock/alpha-crystallin proteins. They enjoy a wide phylogenetic distribution and are important because they function as molecular chaperones, able to mediate many cellular processes through an influence on higher order protein structure. For example, molecular chaperones assist in the transport of proteins into mitochondria and chloroplasts, as well as influencing clathrin lattice dynamics, viral replication and transcriptional activation. Under conditions of stress, some molecular chaperones prevent denaturation of proteins while others may dissociate protein aggregates, refolding monomers derived therefrom or directing their proteolytic destruction. We present in this review an analysis of the emerging literature on the relationship between molecular chaperones and the cytoskeleton, a collection of polymeric structures consisting of microtubules, microfilaments and intermediate filaments. A recent development in this field is identification of the TCP-1 complex as the eukaryotic cytoplasmic chaperonin which directs folding of cytoskeletal proteins such as alpha/beta/gamma-tubulin, actin and centractin. Moreover, the TCP-1 complex is a centrosomal component, apparently involved in the nucleation of microtubules. Other molecular chaperones recognize one or more cytoskeletal elements and in most cases they modulate the assembly of and/or provide protection for their constituent proteins. For example, HSP70 protects the centrosome and perhaps intermediate filaments during heat shock, and like HSP90, it binds to microtubules. Small heat shock proteins interact with microfilaments and intermediate filaments, affect their polymerization and guard them from heat shock by a phosphorylation-dependent mechanism. We conclude that molecular chaperones have different but cooperative roles in the formation and function of the eukaryotic cell cytoskeleton. PMID- 9224762 TI - Modulation of protein synthesis relative to DNA synthesis alters the timing of differentiation in the protozoan parasite Theileria annulata. AB - The control of differentiation through time is critical for the correct ordering of sequential developmental events. A timing mechanism based on the number of mitotic divisions has been proposed for both higher eukaryote and protozoan parasite cellular differentiation. However, the mitotic clock model has not been validated by experiments which altered the proliferation rate of cells in vitro. This study has used the drugs aphidicolin and oxytetracycline to investigate the modulation of differentiation in the protozoan parasite Theileria annulata. The results showed that the timing of macroschizont to merozoite differentiation correlated with expression levels of a merozoite surface antigen during the reversible phase of the differentiation process. In addition, analysis of the effect of the drugs and elevation of culture temperature indicated that altered timing of differentiation was associated with changes to the rate of protein synthesis relative to DNA synthesis. From these results we postulate that the differentiation clock runs on the basis of a progressive elevation of a regulator(s) of merozoite gene expression to a quantitative commitment threshold. We also propose that this mechanism of timing can be corrupted by modulation of the proliferation potential (DNA synthesis) and/or growth potential (factor production) of the cell. The relevance of this model to differentiation in vivo and to other eukaryotic systems is discussed. PMID- 9224763 TI - Microtubule-dependent transport of secretory vesicles visualized in real time with a GFP-tagged secretory protein. AB - Biosynthetic transport from the trans-Golgi network (TGN) to the plasma membrane (PM) is mediated by secretory vesicles. We analyzed secretory vesicle transport in real time using a GFP-tagged secretory protein, hCgB-GFP, consisting of human chromogranin B (hCgB) and green fluorescent protein (GFP). The fusion protein was expressed transiently in Vero cells or in a stable clone after induction with butyrate. After arrest of the biosynthetic protein transport at 20 degrees C, fluorescent hCgB-GFP colocalized with TGN38, a marker of the TGN. Subsequent release of the secretion block at 37 degrees C led to the formation of green fluorescent vesicles. Confocal analysis revealed that these vesicles were devoid of TGN38 and of Texas Red-coupled transferrin and cathepsin D, markers of the endosomal/lysosomal pathway. As determined by fluorometry and metabolic labelling hCgB-GFP was secreted from the TGN to the PM with a t(1/2) of 20-30 minutes. Video-microscope analysis of green fluorescent vesicles showed brief periods of rapid directed movement with maximal velocities of 1 microm/second. Vesicle movement occurred in all directions, centrifugal, centripetal and circumferential, and 50% of the vesicles analyzed reversed their direction of movement at least once within an observation period of 45 seconds. In the presence of nocodazole the movement of fluorescent vesicles ceased. Concomitantly, secretion of hCgB-GFP was slowed but not completely blocked. We suggest that microtubules (MT) facilitate the delivery of secretory vesicles to the PM by a stochastic transport, thereby increasing the probability for a vesicle/target membrane encounter. PMID- 9224764 TI - Leukosialin (CD43, sialophorin) redistribution in uropods of polarized neutrophils is induced by CD43 cross-linking by antibodies, by colchicine or by chemotactic peptides. AB - We investigated a possible association of leukosialin (CD43), the major surface sialoglycoprotein of leukocytes, with neutrophil cytoskeleton. We first analysed the solubility of CD43 in Triton X-100 and observed that CD43 of resting neutrophils was mostly soluble. The small proportion of CD43 molecules, which 'spontaneously' precipitated in Triton, appeared associated with F-actin, as demonstrated by the fact that this insolubility did not occur when cells were incubated with cytochalasin B or when F-actin was depolymerized with DNase I in the Triton precipitate. Cell stimulation with anti-CD43 mAb (MEM59) enhanced this CD43-cytoskeleton association. By immunofluorescence as well as by electron microscopy, we observed a redistribution of CD43 on the neutrophil membrane, initially in patches followed by caps, during anti-CD43 cross-linking at 37 degrees C. This capping did not occur at 4 degrees C and was inhibited by cytochalasin B and by a myosin disrupting drug butanedione monoxime, thus providing evidence that the actomyosin contracile sytem is involved in the capping and further suggesting an association of CD43 with the cytoskeleton. Some of the capped cells exhibited a front-tail polarization with CD43 caps located in the uropod at the rear of the cell. Surprisingly, colchicine and the chemotactic factor fNLPNTL which induce neutrophil polarization associated with cell motility, also resulted in a clustering of CD43 in the uropod, independently of a cross-linking of the molecule by mAbs. An intracellular redistribution of F actin, mainly at the leading front and of myosin in the tail, was observed during CD43 clustering induced by colchicine and in cells polarized by anti-CD43 mAbs cross-linking. We conclude that neutrophil CD43 interacts with the cytoskeleton, either directly or indirectly, to redistribute in the cell uropod under antibodies stimulation or during cell polarization by colchicine, thus highly suggesting that CD43 may be involved in cell polarization. PMID- 9224765 TI - Functional expression of the alpha 7 integrin receptor in differentiated smooth muscle cells. AB - Expression of the alpha7 integrin is developmentally regulated and is thought to be tissue-specific for both skeletal and cardiac muscles. We now report that alpha7 is also strongly and ubiquitously expressed by various types of smooth muscle, including vascular, gastrointestinal and genitourinary smooth muscles. In addition, alpha7 was surface-expressed by a number of smooth muscle cell lines that maintained their differentiated phenotype following adaptation to culture. Studies with the mouse 9E11G smooth muscle cell line showed that the alpha7 integrin mediated both adhesion and motility of these cells on laminin 1 substrates. Alpha7 expression appears to correlate with the smooth-muscle differentiated phenotype. The multipotential P19 mouse embryonic stem cell line lacks alpha7 but uses the alpha6 integrin to adhere to laminin 1. Following retinoic acid-induced P19 differentiation predominantly to the smooth muscle cell lineage, high expression of alpha7 was detected along with partial dependence on alpha7 for binding to laminin. The expression of alpha7 paralleled the induction of smooth-muscle-specific alpha-actin, as revealed by dual-labeling flow cytometry. In contrast, alpha7, which initially was highly expressed on the surface of vascular smooth muscle cell explants, was rapidly downregulated in smooth muscle cell outgrowths as they dedifferentiated into their synthetic phenotype. The results indicate that the expression of alpha7 integrin in smooth muscle cells is associated with their differentiated phenotype and mediates their interaction with laminins. PMID- 9224766 TI - Nuclear envelope assembly in Xenopus extracts visualized by scanning EM reveals a transport-dependent 'envelope smoothing' event. AB - We analyzed the pathway of nuclear envelope assembly in Xenopus egg extracts using field emission in-lens scanning electron microscopy. The binding, fusion, and flattening of vesicles onto the chromatin surface were visualized in detail. The first nuclear pore complexes assembled in flattened patches of nuclear envelope, before the chromatin was fully enclosed by membranes. Confirming previous transmission electron microscope observations, two morphologically distinct types of vesicles contributed to the nuclear membranes: ribosome carrying ('rough') vesicles, many of which bound directly to chromatin, and 'smooth' vesicles, which appeared to associate primarily with other nuclear vesicles or membrane patches. The presence of ribosomes, an outer nuclear membrane marker, on many chromatin-binding vesicles suggested that chromatin attachment proteins integral to the inner membrane were present on vesicles that also carried markers of the outer membrane and endoplasmic reticulum. Chromatin associated vesicles also carried pore membrane proteins, since pore complexes formed when these vesicles were incubated with cytosol. A change in nuclear envelope morphology termed 'envelope smoothing' occurred 5-15 minutes after enclosure. Nuclear envelopes that were assembled in extracts depleted of wheat germ-agglutinin-binding nucleoporins, and therefore unable to form functional pore complexes, remained wrinkled, suggesting that 'smoothing' required active nuclear transport. Lamins accumulated with time when nuclei were enclosed and had functional pore complexes, whereas lamins were not detected on nuclei that lacked functional pore complexes. Very low levels of lamins were detected on nuclear intermediates whose surfaces were substantially covered with patches of pore complex-containing envelope, suggesting that pore complexes might be functional before enclosure. PMID- 9224767 TI - Association of the gamma12 subunit of G proteins with actin filaments. AB - Recent studies have suggested an association between heterotrimeric G proteins, which play a major role in transmembrane signal transduction, and intracellular components. We therefore examined the subcellular localization of isoforms of G protein gamma subunits in Swiss 3T3 and C6 glioma cells, mainly containing the gamma5 and gamma12 subunits. Immunocytochemical double staining with phalloidin showed co-localization of the gamma12 subunit with actin filaments (F-actin), while the gamma5 co-localized with vinculin, suggesting an association with focal adhesion. Pretreatment of cells with Triton X-100 eliminated the gamma5 but not the gamma12 staining. Co-localization of gamma12 and F-actin was preserved when F actin was disorganized with cytochalasin D or reorganized using fetal calf serum. Large amounts of gamma12 were recovered in the vimentin- and tubulin-free F-actin rich fraction prepared from crude cytoskeleton preparations by double depolymerization-repolymerization. Co-localization of Gi2alpha, beta and gamma12 in the F-actin-rich fraction suggested the existence of gamma12 as a betagamma or heterotrimeric complex. Furthermore, purified betagamma12 was found to associate with F-actin in vitro more tightly than betagamma5. These results strongly suggest that the gamma12 subunit associates with F-actin in cells. The observed differential distribution of gamma12 and gamma5 implies functional differences for the two gamma subunits. PMID- 9224768 TI - Concerted action of tenascin-C domains in cell adhesion, anti-adhesion and promotion of neurite outgrowth. AB - We used a new approach to identify domains of chicken tenascin-C required for interaction with cells. Instead of expressing the parts of interest, we deleted them from an otherwise intact tenascin-C molecule and scored for the concomitant change in activity. As a starting point for all mutant constructs we expressed the smallest naturally occurring tenascin-C splice variant in vertebrate cells. The tenascin-C mutants had either deletions of all EGF-like repeats, all fibronectin type III repeats or of the fibrinogen globe. In double mutants the fibronectin type III repeats were deleted together with either the EGF-like repeats or the fibrinogen globe, respectively. All tenascin-C variants assembled correctly to hexameric molecules of the expected molecular characteristics. Intact tenascin-C and the mutant missing the fibrinogen globe did not promote adhesion of chick embryo fibroblasts, whereas both, the hexamers containing solely the fibrinogen globe or the EGF-like repeats were adhesive substrates and even supported cell spreading. When tenascin-C was added to the medium of fibroblasts plated on fibronectin-coated wells, cell adhesion was blocked by intact tenascin-C, but not by mutants missing the fibrinogen globe. In neurite outgrowth assays using dorsal root ganglia, processes formed on all substrates except on the mutant missing only the fibrinogen globe, where the ganglia failed to adhere. The mutants missing the fibronectin type III repeats allowed more rapid neurite outgrowth than all other tenascin-C variants and the mutant consisting essentially of oligomerized EGF-like repeats was as active a substrate for neurite outgrowth as laminin. From the combined data, it is concluded that the activities of intact tenascin-C cannot be mimicked by investigating domain by domain, but the concerted action of several domains leads to the diverse cellular responses. PMID- 9224770 TI - Evidence for colocalization and interaction between 37 and 39 kDa isoforms of secretory carrier membrane proteins (SCAMPs). AB - Secretory carrier membrane proteins (SCAMPs) are proteins of post-Golgi recycling carriers, including regulated secretory organelles. The two major size variants, SCAMP1 (37 kDa) and SCAMP2 (39 kDa), extensively colocalize in membranes of fibroblasts and parotid acinar cells based on immunocytochemistry and velocity centrifugation, although the relative amounts of each variant may differ in selected organelles. SCAMP1, and to a lesser extent, SCAMP2, are substrates for chemical crosslinking in situ, and the recognizable crosslinking products of SCAMP1 suggest potential formation of homomultimers. SCAMP1 and SCAMP2 can be co immunoprecipitated following detergent solubilization, using antibodies that specifically react with only one of the variants. Both the localization and interactions of SCAMPs are reiterated using transfected SCAMP1 that is epitope tagged (myc) at either the NH2 or COOH terminus and an anti-myc antibody. Like other transport vesicle membrane proteins, SCAMPs form complexes that apparently include homomultimers. Furthermore, these studies suggest that both SCAMP1 and SCAMP2 may function together in a single protein complex. PMID- 9224769 TI - The anaphase-promoting complex is required in G1 arrested yeast cells to inhibit B-type cyclin accumulation and to prevent uncontrolled entry into S-phase. AB - Inactivation of B-type cyclin dependent kinases due to ubiquitin-mediated cyclin proteolysis is necessary for the exit from mitosis. In Saccharomyces cerevisiae, proteolysis is initiated at the onset of anaphase and remains active until Cln1 and Cln2 cyclins appear in late G1 of the subsequent cell cycle. A large particle called the anaphase-promoting complex (APC) which is composed of the TPR proteins Cdc16p/Cdc23p/Cdc27p and other proteins is required for B-type cyclin ubiquitination in both anaphase and during G1 phase. The APC has an essential role for the separation of sister chromatids and for the exit from mitosis, but until now it was unclear whether the persistence of APC activity throughout G1 had any physiological role. We show here that the APC is needed in G1 arrested cells to inhibit premature appearance of B-type cyclins and to prevent unscheduled initiation of DNA replication. When pheromone arrested cells of cdc16 and cdc23 mutants were shifted to the restrictive temperature, they underwent DNA replication in the presence of pheromone. DNA replication also occurred in a G1 arrest induced by G1 cyclin (Cln) depletion, indicating that mutant cells with a defective APC initiate DNA replication without the Cln G1 cyclins, which are normally needed for the onset of S-phase. Degradation of Clb2p, Clb3p and Clb5p depends on the APC. This suggests that accumulation of any one of the six B-type cyclin proteins could account for the precocious replication of cdc16 and cdc23 mutants. PMID- 9224772 TI - Quantitative brain microdialysis study on the mechanism of quinolones distribution in the central nervous system. AB - Brain interstitial fluid (ISF) concentrations, which regulate the toxicodynamic effect of quinolone antimicrobial agents (quinolones) in the central nervous system, have been determined for norfloxacin, ofloxacin, fleroxacin, and pefloxacin using a quantitative brain microdialysis technique. Steady-state brain ISF concentrations of the quinolones were 7-30 times lower than the unbound serum concentrations due to restricted distribution in the brain. Cerebrospinal fluid concentrations of the quinolones were approximately twice as high as the brain ISF concentrations, except for norfloxacin. Thus, it seems that an active efflux transport system across the blood-brain barrier is responsible for maintaining brain ISF concentrations lower than unbound serum concentrations at steady-state. A good correlation was observed for norfloxacin, ofloxacin, fleroxacin, and pefloxacin between brain ISF concentrations and total brain concentrations. Moreover, a relatively small difference was observed among the quinolones for the in vitro brain slice-to-medium concentration ratio, compared with an 11-fold difference in the in vivo brain-to-unbound serum concentration ratio after intravenous infusion. These results indicate that the different quinolones studied all exhibit similar apparent binding and/or uptake by brain parenchyma, with an average brain ISF-to-total brain concentration ratio of 0.688. PMID- 9224771 TI - Identification of CYP1A5 as the CYP1A enzyme mainly responsible for uroporphyrinogen oxidation induced by AH receptor ligands in chicken liver and kidney. AB - Uroporphyrinogen is an intermediate of the heme biosynthetic pathway. The oxidation of uroporphyrinogen to uroporphyrin (UROX) has been demonstrated to be catalyzed by mammalian CYP1A2. This reaction has an important role in uroporphyria caused by halogenated aromatic compounds. Two CYP enzymes induced by Ah receptor ligands were purified recently from chick embryo liver. One, designated CYP1A5, was preferentially active in arachidonic acid epoxygenation and the other, designated CYP1A4, in 7-ethoxyresorufin deethylase (EROD) and aryl hydrocarbon hydroxylase (AHH), reactions mainly catalyzed by CYP1A1 in rodents. The amino acid sequences of both CYP1A5 and CYP1A4 are more similar to CYP1A1 than to 1A2, and neither can be classified as an ortholog of mammalian CYP1A1 or 1A2. Here we report that reconstituted purified CYP1A5 was eight times more active than CYP1A4 in catalyzing UROX. The stimulation of UROX by 3,4,3',4' tetrachlorobiphenyl that has been observed in microsomes was also observed with the reconstituted enzymes. Similar dose response relationships were found for induction of UROX and EROD in both chick embryo liver microsomes and in cultured chick hepatocytes, indicating coinduction of CYP1A5 and CYP1A4. UROX was induced by the Ah receptor ligand, 3-methylcholanthrene, in chicken kidney as well as liver. The findings reported here and other evidence that CYP1A4 and CYP1A5 tend to exhibit CYP1A1 and 1A2-like enzyme activites, respectively, indicate that the division of some enzyme activities among CYP1A enzymes applies to different vertebrate classes. PMID- 9224773 TI - Baculovirus-mediated expression and purification of human FMO3: catalytic, immunochemical, and structural characterization. AB - The baculovirus expression vector system was used to overexpress human FMO3 in insect cells for catalytic, structural, and immunochemical studies. Membranes prepared from infected Trichoplusia ni cell suspensions catalyzed NADPH-dependent metabolism of methyl p-tolyl sulfide at rates 20 times faster than those obtained with detergent-solubilized human liver microsomes. Sulfoxidation of the methyl and ethyl p-tolyl sulfides by recombinant human FMO3 proceeded with little stereochemical preference, whereas sulfoxidation of the n-propyl and n-butyl homologs demonstrated increasing selectivity for formation of the (R)-sulfoxide. This chiral fingerprint recapitulated the metabolite profile obtained when detergent-treated human liver microsomes served as the enzyme source. Catalytically active human FMO3 was purified to apparent homogeneity by cholate solubilization and sequential column chromatography on Octyl-Sepharose, DEAE Sepharose, and hydroxyapatite. Purified FMO3 exhibited the same electrophoretic mobility as native microsomal enzyme, and immunoquantitation showed that this isoform represents approximately 0.5% of human liver microsomal protein. Therefore, FMO3 is quantitatively a major human liver monooxygenase. LC/electrospray-mass spectrometry analysis of purified FMO3 identified >70% of the tryptic peptides, including fragments containing motifs for N-linked glycosylation and O-linked glycosylation. Although insect cells have the capacity for glycan modification, MS analysis of the tryptic peptides demonstrated that these sites were not modified in the purified, recombinant enzyme. Edman degradation of the recombinant product revealed that posttranslational modification of human FMO3 by insect cells was limited to cleavage at the N terminal methionine, a process seen in vivo with animal orthologs of FMO3. These studies demonstrate the suitability of this eukaryotic system for heterologous expression of human FMOs and future detailed analysis of their substrate specificities. PMID- 9224774 TI - Cellular uptake of 3H-bestatin in tissues of mice after its intravenous injection. AB - Bestatin, a dipeptide analog, is a potent aminopeptidase inhibitor of bacterial origin. We have previously shown that bestatin inhibits cytosolic exopeptidases in mammalian cells, and results in the accumulation of di- and tripeptide intermediates in cellular protein degradation. Our primary interest is the uptake of bestatin in liver and muscle, 10 min after its intravenous injection into mice. In this short interval, peptide intermediates accumulate linearly in these tissues and permit an estimate of their rates of cellular protein breakdown. Male, CD-1 adult mice received the intravenous injection of 3H-bestatin and 14C sucrose. The disappearance of 3H-bestatin from the plasma, when normalized by the injected radioactivity, was indistinguishable from that of 14C-sucrose. They both drop rapidly during the first 10 min after the injection, followed by a slower exponential disappearance of 3.4% per min, which extrapolates to an apparent volume of distribution of 25 ml/100 g body weight. In two mice, 3 hr after the injection, the urine contained 77.4% and 79.8% of the injected 14C-sucrose, and 70.9% and 73.9% of the injected 3H-bestatin. Other mice were killed 10 min after the injection of 5 mg of bestatin, and the concentration of 3H-bestatin and 14C sucrose was determined in the plasma and various tissues. Using sucrose as a nonpermeant marker of the extracellular space, extracellular 3H-bestatin was calculated and subtracted from the total to estimate the cellular uptake of bestatin. Bestatin was taken up readily in the liver (383-452 microg/g), kidneys (175-191 microg/g), and intestine (137-179 microg/g), but much less in red cells (11 microg/g) or skeletal muscle (4.8 microg/g). Bestatin also entered slowly into erythrocytes in vitro (0.3%/min) by a nonsaturable process. It is suggested that bestatin is taken up through transporter-mediated processes in some cells but not others. PMID- 9224775 TI - In vitro oxidation of famciclovir and 6-deoxypenciclovir by aldehyde oxidase from human, guinea pig, rabbit, and rat liver. AB - Famciclovir, a 9-substituted guanine derivative, is a new antiviral agent which undergoes rapid hydrolysis and oxidation in man to yield the active antiherpes agent, penciclovir. Studies with human liver cytosol have indicated that the oxidation of the penultimate metabolite, 6-deoxypenciclovir, to penciclovir is catalyzed by the molybdenum hydroxylase, aldehyde oxidase. In the present study the oxidation of famciclovir and 6-deoxypenciclovir with partially purified molybdenum hydroxylases from human, guinea pig, rabbit, and rat livers and bovine milk xanthine oxidase has been investigated. Famciclovir and 6-deoxypenciclovir were oxidized predominantly to 6-oxo-famciclovir and penciclovir, respectively, by human, guinea pig, and rat liver aldehyde oxidase. Small amounts of 8-oxo and 6,8-dioxo-metabolites were also formed from each substrate. Famciclovir and 6 deoxypenciclovir were good substrates for rabbit liver aldehyde oxidase but, in each case, two major metabolites were formed. 6-Deoxypenciclovir was converted to penciclovir and 8-oxo-6-deoxypenciclovir in approximately equal quantities; famciclovir was oxidized to 6-oxo-famciclovir and a second metabolite which, on the basis of chromatographic and UV spectral data, was thought to be 8-oxo famciclovir. Two groups of Sprague Dawley rats were identified; those containing hepatic aldehyde oxidase and xanthine oxidase and those with only xanthine oxidase. These have been designated AO-active and AO-inactive rats, respectively. Famciclovir was not oxidized by enzyme from AO-inactive rats or bovine milk xanthine oxidase although 6-deoxypenciclovir was slowly converted to penciclovir by rat liver or milk xanthine oxidase. Inhibitor studies showed in human, guinea pig, and rabbit liver that xanthine oxidase did not contribute to the oxidation of famciclovir and 6-deoxypenciclovir; thus it is proposed that drug activation in vivo would be catalyzed solely by aldehyde oxidase. PMID- 9224776 TI - Identification of the metabolites of the HIV-1 reverse transcriptase inhibitor delavirdine in monkeys. AB - Delavirdine mesylate (U-90152T) is a highly specific nonnucleoside HIV-1 reverse transcriptase inhibitor currently under development for the treatment of AIDS. The metabolism of delavirdine was investigated in male and female cynomolgus monkeys after oral administration of [14C-carboxamide]delavirdine mesylate at single doses of 80 mg/kg and multiple doses of 160 to 300 mg/kg/day. Desalkyl delavirdine was the major metabolite in circulation. In urine, desalkyl delavirdine accounted for nearly half of the radioactivity, with despyridinyl delavirdine and conjugates of desalkyl delavirdine accounting for most of the remaining radioactivity. Bile was mostly composed of desalkyl delavirdine and 6' O-glucuronide delavirdine, with parent drug, 4-O-glucuronide delavirdine, and conjugates of desalkyl delavirdine as significant components. In addition, several minor metabolites were observed in urine and bile of delavirdine treated monkeys. The metabolism of delavirdine in the monkey was extensive and involved N desalkylation, hydroxylation at the C-4' and C-6' positions of the pyridine ring, hydroxylation at the C-4 position of the indole ring, pyridine ring-cleavage, N glucuronidation of the indole ring, and amide bond cleavage as determined by MS and/or one-dimensional and two-dimensional NMR spectroscopies. Phase II biotransformations included glucuronidation, sulfation, and beta-N acetylglucosaminidation. The identification of the N-linked beta-N acetylglucosamine and 4-O-glucuronide metabolites of delavirdine represents novel biotransformation pathways. PMID- 9224778 TI - In vitro identification of the P450 enzymes responsible for the metabolism of ropinirole. AB - The in vitro metabolism of ropinirole was investigated with the aim of identifying the cytochrome P450 enzymes responsible for its biotransformation. The pathways of metabolism after incubation of ropinirole with human liver microsomes were N-despropylation and hydroxylation. Enzyme kinetics demonstrated the involvement of at least two enzymes contributing to each pathway. A high affinity component with a K(M) of 5-87 microM and a low affinity component with a K(M) of approximately two orders of magnitude greater were evident. The high affinity component could be abolished by pre-incubation of the microsomes with furafylline. Additionally, incubation of ropinirole with microsomes derived from CYP1A2 transfected cells readily produced the N-despropyl and hydroxy metabolites. Some inhibition of ropinirole metabolism was also observed with ketoconazole, indicating a minor contribution by CYP3A. Multivariate correlation data were consistent with the involvement of the cytochrome P450 enzymes 1A2 and 3A in the metabolism of ropinirole. Thus, it could be concluded that the major P450 enzyme responsible for ropinirole metabolism at lower (clinically relevant) concentrations is CYP1A2 with a contribution from CYP3A, particularly at higher concentrations. PMID- 9224777 TI - Metabolism of the HIV-1 reverse transcriptase inhibitor delavirdine in mice. AB - Delavirdine mesylate (U-90152T) is a highly specific nonnucleoside HIV-1 reverse transcriptase inhibitor currently under development for the treatment of AIDS. The excretion, disposition, brain penetration, and metabolism of delavirdine were investigated in CD-1 mice after oral administration of [14C]delavirdine mesylate at single doses of 10 and/or 250 mg/kg and multiple doses of 200 mg/kg/day. Studies were conducted with 14C-carboxamide and 2-14C-pyridine labels, as well as 13C3-labeled drug to facilitate metabolite identification. Excretion was dose dependent with 57-70% of the radioactivity eliminated in feces and 25-36% in urine. Pharmacokinetic analyses of delavirdine and its N-desisopropyl metabolite (desalkyl delavirdine) in plasma showed that delavirdine was absorbed and metabolized rapidly, that it constituted a minor component in circulation, that its pharmacokinetics were nonlinear, and that its metabolism to desalkyl delavirdine was capacity limited or inhibitable. Delavirdine did not significantly cross the blood-brain barrier; however, its N isopropylpyridinepiperazine metabolite arising from amide bond cleavage-was present in brain at levels 2- to 3-fold higher than in plasma. The metabolism of delavirdine in the mouse was extensive and involved amide bond cleavage, N desalkylation, hydroxylation at the C-6' position of the pyridine ring, and pyridine ring-cleavage as determined by MS and/or 1H and 13C NMR spectroscopies. N-desalkylation and amide bond cleavage were the primary metabolic pathways at low drug doses and, as the biotransformation of delavirdine to desalkyl delavirdine reached saturation or inhibition, amide bond cleavage became the predominant pathway at higher doses and after multiple doses. PMID- 9224779 TI - Comparative study on formation of hydroxy and sulfur-containing metabolites from different chlorinated biphenyls with 2,5-substitution in rats. AB - 2,4',5-Trichlorobiphenyl (TriCB), 2,3',4',5-tetrachlorobiphenyl (TetraCB), 2,2',4',5,5'-pentachlorobiphenyl (PentaCB), and 2,2',3',4',5,5' hexachlorobiphenyl (HexaCB) were studied with regard to the fecal excretion and tissue distribution of their metabolites after intraperitoneal injection to rats. Major fecal metabolites were 3- and 4-hydroxy and 3- and 4-methylthio derivatives, the substitution ratios depending largely on the degree of chlorination. As the degree of chlorination increased, hydroxy products were more efficiently excreted, whereas the formation of methylthio metabolites greatly decreased. As a result, the excretion ratios of methylthio and hydroxy products varied with 2.8 for TriCB, 1.3 for TetraCB, 0.04 for PentaCB, and 0.02 for HexaCB. The 3-/4-hydroxy substitution ratios were 0.6 for TriCB, 1.4 for TetraCB, 21 for PentaCB, and 35 for HexaCB, whereas the 3-/4-methythio substitution ratios were 1.2 for TriCB, 0.8 for TetraCB, 0.18 for PentaCB, and 0.12 for HexaCB. The formation rate of 3- and 4-methylthio metabolites from each congener was correlated to the accumulation and distribution of 3- and 4-methylsulfonyl derivatives in tissues. The tissue/blood concentration ratios of methylsulfonyl metabolites showed that the 3-methylsulfonyl derivatives from higher chlorinated biphenyls had a relatively high affinity for liver and adipose tissue, whereas the 4-methylsulfonyl derivatives were selectively retained in the lung in all cases. PMID- 9224781 TI - Metabolism and excretion of a new antipsychotic drug, ziprasidone, in humans. AB - The pharmacokinetics, metabolism, and excretion of a new antipsychotic drug, ziprasidone, were studied in four normal male volunteers after oral administration of a single 20 mg dose of a mixture of 14C- and 3H-labeled ziprasidone. Blood, urine, and feces were collected at various intervals for determination of total radioactivity and metabolic profiles. Eleven days after the dose, 20.3 +/- 1% of the administered radioactivity was recovered in the urine and 66.3 +/- 4.8% in feces. The absorption of ziprasidone was rapid, and the C(max) for ziprasidone and metabolites occurred at 2 to 6 hr postdose. Mean peak serum concentration of unchanged drug was 45 ng/ml and a mean AUC(0-t) of 335.7 ng x hr/ml. Mean peak serum concentration of total radioactivity (average of 3H and 14C) was 91 ng-eq/ml and a mean AUC(0-t) of 724.6 ng-eq x hr/ml. On the basis of AUC(0-t) values, approximately 46% of circulating radioactivity was attributable to unchanged drug. Ziprasidone was extensively metabolized and only a small amount (<5% of the administered dose) was excreted in urine and feces as unchanged drug. Twelve metabolites in human urine and serum were identified by ion-spray LC/MS and LC/MS/MS with simultaneous monitoring of radioactivity. The major urinary metabolites were identified as oxindole-acetic acid and its glucuronide conjugate, benzisothiazole-3-yl-piperazine (BITP), BITP-sulfoxide, BITP-sulfone and its lactam, ziprasidone-sulfoxide, and sulfone similar to those identified in rats. In addition, two novel metabolic pathways (reductive cleavage and N-dearylation of the benzisothiazole ring) were identified for ziprasidone in humans. The metabolites resulted by these pathways were characterized as S-methyl dihydroziprasidone, S-methyl-dihydro-ziprasidone sulfoxide, and 6-chloro-5-(2 piperazin-1-yl-ethyl)-1,3-dihydro-indol-2-one, respectively. Ziprasidone sulfoxide and sulfone were the major metabolites in human serum. The affinities of the sulfoxide and sulfone metabolites for 5-HT2 and D2 receptors are low with respect to ziprasidone, and are thus unlikely to contribute to its antipsychotic effects. Structures of the major metabolites were confirmed by chromatographic and spectroscopic comparisons to synthetic standards. Based on the structures of these metabolites, four routes of metabolism of ziprasidone were identified: 1) N dealkylation of the ethyl side chain attached to the piperazinyl nitrogen, 2) oxidation at sulfur resulting in the formation of sulfoxide and sulfone, 3) reductive cleavage of the benzisothiazole moiety, and 4) hydration of the C=N bond and subsequent sulfer oxidation or N-dearylation of the benzisothiazole moiety. The identified metabolites accounted for >90% of total radioactivity recovered in urine. PMID- 9224782 TI - Human and rat lung biotransformation of cyclosporin A and its derivatives using slices and bronchial epithelial cells. AB - Lung biotransformation of the immunosuppressants, cyclosporin A (CSA), the hydroxyethyl derivative SDZ IMM 125 (IMM), and the methylcarbonate derivative SDZ SCP 764 (SCP), was demonstrated in slices from human and rat. The major biotransformation pathway for CSA and IMM (0.1-10 microM) was hydroxylation at amino acid 1 to form AM1 or IMM1, while for SCP it was an esterase cleavage of the methylcarbonate group to form AM1 in both species. The initial rate (0-1 hr) of human total metabolite formation increased proportionally with substrate concentration. AM1 formation was five times greater from SCP, an esterase pathway, than CSA, an oxidative pathway which was inhibited (50%) by ketoconazole. At 24 hr human lung CSA metabolite formation was greater than IMM (3-fold) or SCP (2-fold), whereas rat lung and liver and human bronchial epithelial cell SCP metabolite formation generally exceeded CSA or IMM metabolism. CSA biotransformation is expected to occur throughout the human lung as demonstrated by the similar metabolite profile and extent of metabolism by slices derived from five different regions. The scaling of slice total metabolism to organ metabolism revealed that initially lung CSA metabolite formation would be equal to liver but with time liver metabolism would exceed lung for human and rat. This study has demonstrated that human and rat lung are metabolically active, exhibiting oxidative and esterase pathways toward cyclosporin derivatives. The lung will play an important role in this metabolism, particularly when administered via inhalation; however, the liver will also be a major organ involved in the total clearance of these compounds. PMID- 9224780 TI - Evaluation of omeprazole and lansoprazole as inhibitors of cytochrome P450 isoforms. AB - The human clearance of omeprazole and lansoprazole is conducted primarily by the hepatic cytochrome P450 (CYP) system. Efficacy data indicate few differences between these two drugs, but they may exhibit discrete drug interaction profiles. To compare the potency and specificity of these drugs as inhibitors of CYP isoforms, we performed in vitro studies with human liver microsomal preparations. Both drugs were potent, competitive inhibitors of CYP2C19, as measured by the conversion of S-mephenytoin to 4-hydroxymephenytoin (k(i) = 3.1 +/- 2.2 microM for omeprazole, K(i) = 3.2 +/- 1.3 microM for lansoprazole). For omeprazole, the highest concentration at which >70% inhibition of CYP2C19 was observed with no significant inhibitory effect on other isoforms was at least 20 times greater than K(i). Both drugs were competitive inhibitors of CYP2C9-catalyzed conversion of tolbutamide to 4-hydroxytolbutamide (K(i) = 40.1 +/- 14.8 microM for omeprazole, K(i) = 52.1 +/- 1.4 microM for lansoprazole) and were noncompetitive inhibitors of CYP3A-catalyzed conversion of dextromethorphan to 3 methoxymorphinan (K(i) = 84.4 +/- 4.0 microM for omeprazole, K(i) = 170.4 +/- 7.1 microM for lansoprazole). Lansoprazole was at least 5 times more potent (K(i) = 44.7 +/- 22.0 microM) than omeprazole (k(i) = 240.7 +/- 102.0 microM) as an inhibitor of CYP2D6-mediated conversion of dextromethorphan to dextrorphan. No inhibition of CYP1A2, assessed by measuring the conversion of phenacetin to acetaminophen, was noted. Our data suggest that whereas the inhibitory profiles of these two drugs are similar, lansoprazole may be the more important in vitro inhibitor of CYP2D6. Since its inhibition is very potent and has a broad "window of selectivity," omeprazole seems to be a useful, selective inhibitor of CYP2C19. PMID- 9224783 TI - Disposition of [14C]avitriptan in rats and humans. AB - Avitriptan is a new 5-HT1-like agonist with abortive antimigraine properties. The study was conducted to characterize the pharmacokinetics, absolute bioavailability, and disposition of avitriptan after intravenous (iv) and oral administrations of [14C]avitriptan in rats and oral administration of [14C]avitriptan in humans. The doses used were 20 mg/kg iv and oral in the rat, 10 mg iv in humans, and 50 mg oral in humans. The drug was rapidly absorbed after oral administration, with peak plasma concentrations occurring at 0.5 hr postdose. Absolute bioavailability was 19.3% in rats and 17.2% in humans. Renal excretion was a minor route of elimination in both species, with the majority of the dose being excreted in the feces. After a single oral dose, urinary excretion accounted for 10% of the administered dose in rats and 18% of the administered dose in humans, with the remainder excreted in the feces. Extensive biliary excretion was observed in rats. Avitriptan was extensively metabolized after oral administration, with the unchanged drug accounting for 32% and 22% of the total radioactivity in plasma in rats and humans, respectively. Plasma terminal elimination half-life was approximately 1 hr in rats and approximately 5 hr in humans. The drug was extensively distributed in rat tissues, with a tendency to accumulate in the pigmented tissues of the eye. PMID- 9224784 TI - Glucuronidation of retinoids by rat recombinant UDP: glucuronosyltransferase 1.1 (bilirubin UGT). AB - Rat liver recombinant BR1UGT1.1 was found to have significant activity toward retinoid substrates. UGT1.1 glucuronidation activity was 91 +/- 18 pmol/mg x min for atRA and 113 +/- 19 pmol/mg x min for 5,6-epoxy-atRA. The apparent K(M) and V(max) of atRA acid glucuronidation by UGT1.1 were 59.1 +/- 5.4 microM and 158 +/ 43 pmol/mg x min, respectively. SDS-PAGE and Western blot analysis of UGT1.1 transfected HK293 membrane proteins photolabeled with [11,12-3H]atRA revealed a protein of approximately 56 kDa that was labeled by [3H]atRA, detected by anti pNP UGT antibody and not present in membranes from nontransfected HK293 cells. Liver microsomes from Gunn rats, which lack UGT1.1, had significant activity toward atRA (111 +/- 28 pmol/mg x min). PMID- 9224785 TI - Decay rates of anti-HIV dideoxynucleotides in tissue culture systems: a simple correction for the effect of cell replication. AB - Measurement of intracellular drug levels in cell culture systems can be of predictive value in establishing rational clinical dosage schedules. Such in vitro measurements carried out with anti-HIV agents of the 2',3' dideoxynucleoside (ddN) class have shown that many of the pharmacologically active ddNTP metabolites of these agents have relatively long intracellular half lives and little or no host-cell cytotoxicity. As a consequence, replication of drug-exposed cells continues at an unperturbed rate so that a systematic dilution error occurs in the measurement of ddNTP decay half-times. The aim of this study is to present a simple general formulation for the correction of measured t1/2 values for ddNTPs and for other agents with similar intracellular pharmacokinetic properties. Two factors of practical interest emerge: first, the error is greater for agents with slow intracellular clearance rates than for agents with rapid rates; and second, for cell lines with long doubling times, the measured t1/2 values approach more closely to the true t1/2-values, until with the extreme case (quiescent or "G(o)" cells), the observed and true decay times are identical. The greatest dilution errors are seen with adenodine-based agents such as ddATP and 2'-F-ddATP, while the smallest errors are seen with rapidly cleared agents of the dideoxythymidine class. PMID- 9224786 TI - Characterization of the novel benzisothiazole ring-cleaved products of the antipsychotic drug ziprasidone. AB - Characterization of two novel benzisothiazole ring cleaved metabolites of the antipsychotic drug, ziprasidone (ZIP), in rat has been described. Metabolites designated M6 and M9 were isolated from urine and bile of the rat dosed with radiolabeled ZIP and purified by reversed phase HPLC. The chemical structures of these metabolites were assigned based on tandem mass spectrometry in combination with chemical derivatization techniques. M6 and M9 were unaffected upon treatment with N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide. Reaction of M9 with aqueous TiCl3 also did not change the HPLC retention time or the CID spectrum of metabolite M9. These data excluded the possibility that these metabolites were owing to N-oxidation and/or aromatic hydroxylation. M6 and M9 were generated only when in vitro incubations of ZIP were conducted with human liver S-9 fraction in the presence of S-adenosyl-L-methionine. Based on these data, metabolites M6 and M9 were identified as S-methyl-dihydro-ZIP and S-methyl-dihylro-ZIP-sulfoxide, respectively. The structure of M9 was unambiguously confirmed by comparing the LC/MS retention time and mass spectral data with synthetic standard. A mechanism for the formation of these metabolites from ZIP is proposed. PMID- 9224787 TI - Distribution of N(omega)-nitro-L-arginine following topical and intracerebroventricular administration in the rat. AB - The distribution of the nitric oxide synthase inhibitor, N(omega)-nitro-L arginine (L-NA), in the rat brain following either topical or intracerebroventricular application was examined using 14C-labeled L-NA. Two hours after topical application, the concentration of L-NA 1 mm below the surface was approximately 20% of the concentration at the surface, and less than 3% at 2 mm below the cortical surface. L-NA infused into the lateral cerebral ventricle distributes only in regions in close proximity to the ventricle even 8 h after administration. Consequently, intracerebroventricular administration of nitric oxide synthase inhibitors may not be useful if global inhibition of nitric oxide synthase is desired. PMID- 9224788 TI - The modulation of sensorimotor gating deficits by mesolimbic cholecystokinin. AB - The effects of cholecystokinin (CCK) in an animal model of sensorimotor-gating deficits with strong face, construct and predictive validity for schizophrenia were investigated. Prepulse inhibition (PPI) occurs when a weak acoustic lead stimulus inhibits the startle response to a loud startling stimulus. Infusions of sulfated CCK-8 in the posterior nucleus accumbens potentiated apomorphine-induced disruption of PPI but had no effect on baseline PPI or the amplitude of acoustic startle reflex itself. The results provide evidence that mesolimbic CCK may play a role in regulating sensorimotor gating deficits but contradict earlier notions that CCK agonists may have antipsychotic properties and upon which clinical trials of CCK agonists in schizophrenia were based. Rather, these results suggest that antagonists of CCK may display neuroleptic-like actions on deficits in PPI and may hold greater promise as antipsychotics. PMID- 9224789 TI - Propofol potentiates the depressant effect of alfentanil in isolated neonatal rat spinal cord and blocks naloxone-precipitated hyperresponsiveness. AB - Our previous studies have shown that a benzodiazepine potentiates opioid actions on spinal cord by blocking a hyperresponsiveness that may be related to the development of opioid tolerance and withdrawal. The present study was designed to test whether propofol, which like benzodiazepines acts on GABA(A) receptors, displays similar interactions with opioids. Spinal cords isolated from 1-7 day old rats were arranged to record the slow ventral root potential (sVRP) elicited by stimulating a lumbar dorsal root. A concentration of propofol which by itself did not depress sVRP significantly enhanced the apparent potency of alfentanil and blocked the increase in sVRP observed when alfentanil is followed by naloxone. The results suggest that enhancement of GABA inhibition may increase opioid potency by inhibiting the development of acute tolerance. PMID- 9224790 TI - An atypical diencephalic nucleus in actinopterygian fishes: visual connections and sporadic phylogenetic distribution. AB - Nucleus rostrolateralis is a distinctive diencephalic nucleus in some ray-finned fishes. In the osteoglossomorph Pantodon, it is a large ovoid nucleus with visual system connections. A topologically and cytoarchitectonically similar nucleus has been found in six species of non-osteoglossomorph fishes, two species of gars and four euteleosts. Of the latter, two are ostariophysans of the genus Danio, one an atherinomorph, and one a notothenioid percomorph. A variety of characteristic similarities can be found in nucleus rostrolateralis among all these species. The present study reports on these similarities in Danio and in the euteleost Xiphophorus as compared with the nucleus in Pantodon. While this nucleus has a phylogenetic distribution that might imply convergent evolution, the high degree of similarity in its features across species strongly suggests that its genetic basis may be the same despite the lack of phenotypic homology. PMID- 9224791 TI - Synaptically coupled central nervous system neurons lack centrosomal gamma tubulin. AB - In cycling cells, microtubule assembly is initiated at the centrosome and requires the centrosomal protein gamma-tubulin. Previously, it was reported that gamma-tubulin is present at the centrosome of cervical ganglion cells undergoing axonal growth, but not in the axons or dendrites. We find that although gamma tubulin is present at the centrosomes of neurons just beginning to extend processes, it is not associated with centrosomes in hypothalamic and cortical neurons on which functional synaptic connections have formed. In contrast, another centrosomal protein, pericentrin, is associated with the centrosome at all stages. These results suggest that centrosomal microtubule nucleation is required for early stages of neurogenesis to supply sufficient microtubule polymer to support rapid axonal growth, but is not required for maintenance of axonal microtubules in synaptically coupled neurons. PMID- 9224792 TI - Densitometric evaluation of markers for cholinergic transmission in rat superior olivary complex. AB - Cholinergic transmission in the rat superior olivary complex (SOC) was assessed by densitometric measurements of labeling with several markers: acetylcholinesterase (AChE) histochemistry, choline acetyltransferase (ChAT) immunohistochemistry, and muscarinic acetylcholine receptor (M35) immunohistochemistry and N-methylscopolamine (NMS) binding autoradiography, as well as its subtype 2 (m2) immunohistochemistry. The markers which may occur in cholinergic neurons (ChAT, m2 receptor and AChE) showed dense labeling in the ventral nucleus of the trapezoid body (VNTB), in line with other evidence that it contains cholinergic neuron somata. The markers which are predominantly in cholinoceptive neurons (M35 and NMS) showed less labeling in the SOC, suggesting few muscarinic cholinergic inputs. These results are compared with those for the cochlear nucleus. PMID- 9224793 TI - A left temporal lobe impairment of auditory information processing in schizophrenia: an event-related potential study. AB - A measure of auditory prepulse inhibition (PPI) is the reduction of the scalp recorded P1 event-related potential (ERP) after a sound that is preceded by 100 300 ms by a click as prepulse. This measure of sensory gating was adapted to study the effect of a prepulse on processing tones that were part of a 'go no-go' discrimination. ERPs were recorded at right and left, frontal and temporal sites in groups of patients with schizophrenia (SCH) or obsessive compulsive disorder (OCD) and healthy controls (CON). A prepulse 100 ms but not 500 ms before either tone reduced the P1 ERP amplitude in healthy and OCD subjects but not SCH patients. At frontal and temporal recording sites the P1 amplitude was similar bilaterally in controls but showed a right temporal shift in the SCH patients. If the tone was the 'no-go' tone, the prepulse reduced the N1 amplitude in both the CON and SCH groups. The N1 was similar, bilaterally in controls but again showed a right temporal shift in the SCH group. These results show a reduction of a PPI like effect on early processing (P1) that is more marked in the left hemisphere of SCH patients and may affect channel selection for processing information (N1) about task-relevant sounds. PMID- 9224794 TI - Potentiation of synaptic transmission in the rat dentate gyrus in vitro by (S) 3,5-dihydroxyphenylglycine ((S)-DHPG). AB - The direct activation of metabotropic glutamate receptors (mGluRs) by 1S,3R aminocyclopentane dicarboxylate (1S,3R-ACPD), has previously been shown to induce a relatively fast (maximum at 10 min) and slow (90 min) onset long-term potentiation (LTP) of synaptic transmission in the hippocampus. Here we report the first evidence for a relatively fast onset LTP of synaptic transmission in the immature male rat (50-100 g) dentate gyrus in vitro by application of the mGluR type I agonist, (S)-3,5-dihydroxyphenylglycine ((S)-DHPG; 20 microM). Bath application of (S)-DHPG caused a transient depression of the field excitatory postsynaptic potential (EPSP) slope, followed after washout by a relatively rapidly developing potentiation of synaptic transmission to a maximum increase at 12-15 min (161.1 +/- 11.4% compared to controls at 15 min; n = 8). This effect was not observed following perfusion with the enantiomer (R,S)-DHPG at the same dose. The (S)-DHPG potentiation occluded tetanically induced LTP and vice versa. The potentiation was antagonised by the non-specific mGluR antagonist (R,S)-alpha methyl-4-carboxyphenylglycine ((R,S)-MCPG) at high doses (500-1000 microM) but was unaffected in the presence of the N-methyl-D-aspartate (NMDA) receptor blocker, D(-)-2-amino-5-phosphonopentanoic acid (D-AP5; 50 microM). Our results demonstrate a robust NMDA-independent LTP induced by (S)-DHPG that occludes tetanically induced LTP. PMID- 9224795 TI - Interleukin-1beta sensitizes the response of the gastric vagal afferent to cholecystokinin in rat. AB - Interleukin-1beta (IL-1beta) and cholecystokinin (CCK) are important mediators in the development of anorexic response during disease. The role of IL-1beta and CCK in the peripheral mechanisms of anorexia was studied by recording the mass afferent activity of the gastric vagal nerve in anesthetized rats. The i.v. administration of CCK (1 nmol) increased the activity of the vagal nerve, and this response was raised by 55-72% 2 h after i.v. injection of IL-1beta. It is proposed that IL-1beta-induced anorexia is mediated via the sensitization of type A CCK receptors in the periphery. PMID- 9224796 TI - Amyloid beta-induced neurotoxicity is associated with phospholipase D activation in cultured rat hippocampal cells. AB - The role of phospholipase D (PLD) in amyloid beta (Abeta)-induced neurotoxicity was studied by comparing the effects of Abeta (1-40) on PLD activity and release of lactate dehydrogenase (LDH) from cultured rat hippocampal cells. PLD activity was determined in [3H]myristic acid-labeled cells by measuring the formation of [3H]phosphatidylethanol in the presence of ethanol (0.5%), and LDH activity in the cell media was measured via colorimetric assay. Abeta (50 microM), aged for 3 days to allow for peptide aggregation, acutely (1 h) stimulated PLD activity. Unaged Abeta (50 microM) had no acute (1 h) effect on PLD activity, but significantly stimulated PLD activity by 87% when incubated with cells for 1-3 days. Abeta (50 microM)-induced PLD activity was closely correlated with Abeta (50 microM)-induced LDH release over a time course of 1-3 days. These data suggest that PLD activation may be involved in Abeta-induced neurotoxicity. PMID- 9224797 TI - No evidence for specific opioid effects on batrachotoxin-modified sodium channels from human brain synaptosomes. AB - Human central nervous system (CNS) sodium channels modified by batrachotoxin and incorporated inter voltage-clamped lipid bilayers, were exposed to various concentrations of the opioid alfentanil (0.2-8.0 mM). Alfentanil caused a concentration-dependent and membrane potential independent reduction of the single channel amplitude and the fractional channel open-time. The weighted computer fit of the dose-response curve yielded a maximal conductance block of 50% with an EC50 of 1.3 mM. These effects occurred at levels beyond clinically relevant human serum/brain levels (0.003 mM) but within the predicted concentration range using the Meyer-Overton (lipid solubility/anaesthetic potency) correlation. Thus, human CNS sodium channels are probably not a main target site for the clinical effects of alfentanil but they provide a model system to estimate the proportion of the lipophilic interactions contributing to its overall effect. PMID- 9224798 TI - Ultrastructural localization of neuronal nitric oxide synthase-immunoreactivity in the rat ileum. AB - The location of neuronal nitric oxide synthase-immunoreactivity (NOS-IR) in whole mount preparations of muscularis externa of rat ileum was determined by using pre embedding electron microscope immunocytochemistry. Several neurons, nerve fibers and nerve endings in the myenteric plexus (MP) and nerve endings within the muscle layers were found to be NOS-IR. These nerve endings were especially numerous in the deep muscular plexus (DMP) and much closer to interstitial cells of Cajal (ICC) than to smooth muscle cells. Some of the ICC-MP were NOS-IR. These findings indicate that ICC-MP are apparently able to produce NO and ICC-DMP are the ileal ICC type very richly innervated by the NO releasing nerves. PMID- 9224799 TI - Day-night variation of pituitary adenylate cyclase-activating polypeptide (PACAP) level in the rat suprachiasmatic nucleus. AB - Adenylate cyclase-activating polypeptide (PACAP) is synthesized in the retinal ganglion cells which terminate on vasoactive intestinal polypeptide neurons in the suprachiasmatic nucleus (SCN), the location of circadian clock. To examine whether PACAP exhibits daily variations in the rat SCN, we measured endogenous PACAP contents throughout the day under 12:12 h light-dark or constant dark conditions. PACAP level was low during the light periods, high during the dark periods, and was stable under constant dark conditions. In the periventricular nucleus of the hypothalamus and cerebral cortex, PACAP content did not show any significant variation throughout the day. Our findings suggest that PACAP content in the SCN may be changed by lighting conditions. Thus, PACAP-containing neurons may play certain roles in the entrainment of circadian rhythms. PMID- 9224800 TI - Sorbitol accumulation and transmembrane efflux in osmotically stressed JS1 schwannoma cells. AB - Sorbitol accumulation and transmembrane efflux in JS1 schwannoma cells were examined during osmotic stress in the presence of a sorbitol dehydrogenase (SD) inhibitor and following return to iso-osmotic conditions. SD inhibition promoted sorbitol accumulation under hyperglycemic and/or hyperosmotic conditions, and sorbitol efflux during iso-osmotic incubation was prevented by cooling to 4 degrees C or by quinidine. It appears that sorbitol levels in JS1 cells are dependent on SD activity and that sorbitol is rapidly removed upon restoring iso osmotic conditions. PMID- 9224801 TI - Effects of dopamine on N-terminus-deleted human tyrosine hydroxylase type 1 expressed in Escherichia coli. AB - N-Terminus-deleted mutants and wild-type human tyrosine hydroxylase type 1 were expressed in Escherichia coli (E. coli) and utilized to investigate the dopamine induced decrease in the enzyme catalytic activity and also to identify the specific portion in the N-terminus that affects the efficiency of the inhibitory action of dopamine. Supernatants of bacterial lysates were used as enzyme samples. The pH profiles of the enzyme catalytic activity were affected according to the degree of the deletion. The deletion up to 39 amino acid residues was enough to abolish the inhibitory effect of dopamine in the basic pH range. These results suggest that the inhibition by dopamine of tyrosine hydroxylase activity is closely related to the amino acid sequence in the N-terminus of the enzyme. PMID- 9224802 TI - Different effects of Alzheimer-associated mutations of presenilin 1 on its processing. AB - Presenilin 1 (PS 1) shows missense mutations in most early-onset familial Alzheimer's disease (FAD). Transfection of cDNA for wild type PS 1 into rat pheochromocytoma PC12 cells generated a 47 kDa full-size PS 1 protein, which was processed into a 28 kDa N-terminal fragment and a 19 kDa C-terminal fragment. We prepared selected Alzheimer-associated mutations (Gly384Ala, Leu392Val, and Cys410Tyr) of PS 1, which localized after a possible cleavage site. By transient expression in PC12 cells and rat glioma cell line, C6, we examined their influence on the processing of PS 1. Cys410Tyr inhibited proteolytic processing of PS 1, while Gly384Ala and Leu392Val did not. Thus, the Alzheimer related mutations can be divided into two groups in terms of their effect on the proteolytic cleavage of PS 1. PMID- 9224803 TI - Increase of urocortin-like immunoreactivity in the rat supraoptic nucleus after dehydration but not food deprivation. AB - The effects of dehydration and food deprivation on urocortin-like immunoreactivity (Ucn-IR) in the rat supraoptic nucleus (SON) and the paraventricular nucleus (PVN) were examined by immunohistochemistry. Water deprivation for 48 h caused a significant increase in the number of Ucn-IR neurons in the SON, compared with control. Ucn-IR fibers and varicosities in the SON and the internal zone of the median eminence (ME) were increased, but a few and faint Ucn-IR neurons and fibers were observed in the PVN. On the other hand, food deprivation for 48 h caused a significant decrease in the number of Ucn-IR neurons in the SON, compared with control. Ucn-IR fibers and varicosities in the SON and the ME were fewer than those in controls. Ucn-IR neurons and fibers in the PVN were not detected after food deprivation. These results suggest that Ucn in the SON may be involved in the central regulation of water balance and nutrient homeostasis. PMID- 9224805 TI - Colloquium on signaling and molecular structure in pharmacology (La Jolla, California, March 11-12, 1997). PMID- 9224804 TI - Leukemia inhibitory factor (LIF) mRNA-expressing neuronal subpopulations in adult rat basal forebrain. AB - We have previously found leukemia inhibitory factor (LIF) mRNA in neurons of the adult rat brain. To identify which neuronal subpopulations are expressing LIF transcripts, non-radioactive in situ hybridization was combined with immunocytochemistry for various neuronal markers. Studying the rat basal forebrain and cerebral cortex, we find LIF mRNA is expressed in both cholinergic and GABAergic neurons. These data suggest a role for LIF in the function of these mature neurons. PMID- 9224806 TI - High affinity glutamate transporters: regulation of expression and activity. AB - L-Glutamic acid is a major excitatory neurotransmitter in the mammalian central nervous system. The termination of the glutamatergic transmission and the clearance of the excessive, neurotoxic concentrations of glutamate is ensured by a high affinity glutamate uptake system. Four homologous types of Na/K-dependent high affinity glutamate transporters, glutamate/aspartate transporter, glutamate transporter 1, excitatory amino acid carrier 1, and excitatory amino acid transporter 4, have recently been cloned and were assigned to a separate gene family, together with two neutral amino acid carriers, alanine/serine/cysteine transporter 1/serine/alanine/threonine transporter and adipocyte amino acid transporter. The genomic organization of these transporters is still under investigation. Very little is known about the nature of the factors and molecular mechanisms that regulate developmental, regional, and cell type-specific expression of the glutamate transporters and their aberrant functioning in neurodegenerative diseases (e.g., amyotrophic lateral sclerosis and Alzheimer's disease). Some experimental conditions (e.g., ischemia, corticostriatal lesions, hyperosmolarity, culturing conditions) and several naturally occurring and synthetic compounds (e.g., glutamate receptor agonists, dopamine, alpha1- and beta-adrenergic agonists, cAMP, phorbol esters, arachidonic acid, nitric oxide, oxygen free radicals, amyloid beta-peptide, tumor necrosis factor-alpha, glucocorticosteroids, unidentified neuronal factors) affect the molecular expression and activity of glutamate transporters. Further elucidation of the molecular events that link epigenetic signals with transcriptional and post transcriptional mechanisms (e.g., alternative splicing, translation and post translational modifications) is crucial for the development of selective pharmacological tools and strategies interfering with the expression of the individual glutamate transporters. PMID- 9224807 TI - Nonpeptide mimic of bradykinin with long-acting properties at the bradykinin B2 receptor. AB - Kinins, members of a family of peptides released from kininogens by the action of kallikreins, exhibit a variety of biological activities including vasodilation, increased vascular permeability, contraction of smooth muscle cells, and activation of sensory neurons. However, investigation of the physiological actions of kinins has been greatly hampered because its effects are curtailed by rapid proteolysis in blood, lung, and liver. We describe the pharmacological characteristics of a novel nonpeptide bradykinin receptor agonist FR190997 (8 [2,6-dichloro-3-[N-[(E)-4-(N-methylcarbamoyl)cinnamidoacetyl ]-N methylamino]benzyloxy]-2-methyl-4-(2-pyridylmethoxy)quinoli ne). FR190997 markedly stimulated phosphatidylinositol hydrolysis in Chinese hamster ovary cells permanently expressing the human bradykinin B2 receptor. The response of phosphatidylinositol hydrolysis was antagonized by the B2 receptor selective antagonist Hoe 140 (D-Arg-[hydroxyproline3,beta-thienylalanine4,D-Tic7,++ +Oic8]bradykinin). In competitive experiments using membranes prepared from Chinese hamster ovary cells expressing the human bradykinin receptor subtypes, FR190997 showed a high affinity binding to the B2 receptor with IC50 value of 5.3 nM and no binding affinity for the B1 receptor. In vivo, FR190997 mimics the biological action of bradykinin and induces hypotensive responses in rats with prolonged duration. Therefore, FR190997 is a highly potent and subtype-selective nonpeptide agonist which displays high intrinsic activity. This compound should represent a powerful tool for further investigation of the physiology and pathophysiology of bradykinin receptors. PMID- 9224808 TI - Palmitoylation of the V2 vasopressin receptor. AB - Palmitoylation of the V2 vasopressin receptor (V2R) and its functional role were investigated in transfected cells. Palmitoylation was assessed by incubating transfected cells with [3H]palmitic acid and immunoprecipitating the receptor with an antibody raised against a portion of the third intracellular loop of V2R. Wild-type and nonglycosylated V2R yielded tritium signals at 45-55 and 40 kDa, respectively, demonstrating that the V2R is palmitoylated and that receptor palmitoylation is independent of glycosylation. Substitution of CC341/342 for serines eliminated receptor palmitoylation, whereas replacement of a single amino acid, C341S or C342S, restored partial palmitoylation. Saturation binding assays revealed decreased cell surface expression of the nonpalmitoylated receptor compared with the wild-type; this effect was more pronounced when a truncated form of V2R (G345ter) was studied. The presence of either cysteine residue (C341S or C342S) elevated receptor expression to normal levels, most likely due to the partial restoration of palmitoylation. Ligand binding affinity, hormone-induced stimulation of adenylyl cyclase activity, receptor internalization, and desensitization were not affected by the absence of palmitoylation. No increase but rather a slight decrease in the extent of receptor palmitoylation was detected after exposure to vasopressin. It was concluded that the V2R is palmitoylated in both cysteines, each cysteine is palmitoylated independently from the other, and palmitoylation enhances cell surface expression of the V2R. PMID- 9224809 TI - Cytokine gene expression during ontogeny in murine thymus on activation of the aryl hydrocarbon receptor by 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) binds and activates the aryl hydrocarbon receptor (Ah-R), an endogenous transcription factor that is expressed in the thymus. TCDD exposure leads, among other effects, to thymus atrophy and immunosuppression. We previously analyzed the interference of TCDD with differentiation processes in fetal thymus organ cultures and found that in the presence of TCDD, the proliferation rate of immature (CD4- CD8- and CD4- CD8+ HSA+) thymocytes is inhibited, whereas the maturation along the CD4/CD8 path is accelerated. Moreover, the differentiation of thymocytes is skewed by TCDD at < or = 40% (compared with approximately 15% without TCDD) of the CD8 single positive subset of future cytotoxic T cells, and apparently more cells audition for and pass positive selection. The fetal murine thymus expresses functional Ah R mRNA, as shown by reverse transcription-polymerase chain reaction and TCDD inducible CYP1A1 and CYP1B1 expression. Because the differentiation of thymocytes is to a considerable extent controlled by cytokines and many cytokine genes are potential targets of the Ah-R due to Ah-R-binding elements (xenobiotic response elements) in their promoters, we analyzed the cytokine expression in fetal thymus organ culture exposed to TCDD. Fetal thymi were cultured from gestation day 15 for < or = 8 days, thus covering ex vivo the period after population of the thymus anlage until birth. We show with semiquantitative reverse transcription polymerase chain reaction that more interleukin (IL)-1beta, IL-2, IL-6, tumor growth factor (TGF)-beta3, and tumor necrosis factor-alpha are produced in TCDD exposed thymi, whereas other cytokines (e.g., TGF-beta1, PAI-2, or IL-4) are only slightly up- and down-modulated during the culture period or not modulated at all (e.g., IL-1beta, IL-7, interferon-gamma, and TGF-beta2). PMID- 9224810 TI - Role of amino- and carboxyl-terminal regions of G(alphaZ) in the recognition of Gi-coupled receptors. AB - Many Gi-coupled receptors are known to interact with the pertussis toxin (PTX) insensitive Gz protein. Given that the alpha subunits of Gi and Gz share only 60% identity in their amino acid sequences, their receptor-interacting domains must be highly similar. By swapping the carboxyl termini of alpha i2 and alpha z with each other or with those of alpha t, alpha12, and alpha13, we examined the relative contributions of the carboxyl-end 36 amino acids of the alpha chains toward receptor recognition. Chimeric alpha chains lacking the site for PTX catalyzed ADP-ribosylation were coexpressed with the type II adenylyl cyclase (AC II) and one of several Gi-coupled receptors (formyl peptide, dopamine D2, and delta-opioid receptors) in human embryonic kidney 293 cells. The alpha i2/alpha z chimera was able to interact with both aminergic and peptidergic receptors, resulting in betagamma-mediated stimulation of AC II in the presence of agonists and PTX. Functional and mutational analyses of alpha i2/alpha z revealed that this chimera can inhibit the endogenous ACs of 293 cells. Similarly, the alpha z/alpha i2 chimera seemed to retain the abilities to interact with receptors and inhibit cAMP accumulation. Fusion of the carboxyl-terminal 36 amino acids of alpha z to a backbone of alpha t1 produced a chimera, alpha t1/alpha z, that did not couple to any of the Gi-coupled receptors tested. Interestingly, an alpha13/alpha z chimera (with only the last five amino acids switched) displayed differential abilities to interact with receptors. Signals from aminergic, but not peptidergic, receptors were transduced by alpha13/alpha z. A similar construct, alpha12/alpha z, behaved just like alpha13/alpha z. These results indicated that "alpha i-like" or "alpha z-like" sequences at the carboxyl termini of alpha subunits are not always necessary or sufficient for specifying interaction with Gi-coupled receptors. PMID- 9224811 TI - Thyroid regulation of NADPH:cytochrome P450 oxidoreductase: identification of a thyroid-responsive element in the 5'-flank of the oxidoreductase gene. AB - The current study demonstrates that T3-activated transcription of the NADPH:cytochrome P450 oxidoreductase (P450R) gene is dependent on the thyroid hormonal status of the animal, with both transcriptional and post-transcriptional pathways being important in regulating the cellular P450R mRNA level. The region required for transcriptional activation of the P450R gene by T3 has been identified. Nuclear run-on experiments demonstrated that the effects of T3 on P450R transcription are dependent on thyroid status, with a transcriptional enhancement obtained in T3-treated hypothyroid rat liver (1.8-fold increase) but not in T3-treated euthyroid animals. Transient cotransfection of P450R promoter/chloramphenicol acetyl transferase (CAT) constructs and the thyroid hormone receptor beta1 (TR beta1) expression plasmid into rat hepatoma H4IIE cells resulted in a 2.4-fold induction of promoter activity that was both T3 and TR beta1 dependent. Analysis of promoter deletion constructs identified a P450R thyroid response region (P450R-TRE; bases, -564 to -536) containing three imperfect direct repeats of the thyroid response motif, AGGTCA. Mutational analysis further established that T3 induction was dependent only on the upstream direct repeat, having the sequence AGGTGAgctgAGGCCA. Footprint analysis showed that all three motifs were protected by proteins present in rat liver nuclear extracts, and a direct interaction between P450R-TRE and T3 receptors TR alpha1 and TR beta1 was demonstrated by gel-shift analysis. In vitro binding studies with P450R-TRE revealed the formation of heterodimeric complexes when TR alpha1 was coincubated with either the retinoic X receptor alpha or nuclear extract from rat liver, COS, or H4IIE cells. In addition, placement of the P450R-TRE upstream of the T3-nonresponsive heterologous thymidine kinase promoter resulted in a 2.7 fold transcriptional enhancement that was both T3 and TR beta1 dependent. Previous studies have demonstrated that T3 augments P450R mRNA levels approximately 20-30-fold and approximately 12-fold, respectively, in hypothyroid and euthyroid rats. Hence, for the hypothyroid state, transcriptional and post transcriptional events contribute to the T3-induced mRNA increases; however, the marked increase in message level in T3-treated euthyroid animals depends primarily on post-transcriptional pathways. PMID- 9224812 TI - Recombinant advanced glycation end product receptor pharmacokinetics in normal and diabetic rats. AB - Vascular dysfunction in patients with diabetes mellitus is related to advanced glycation end product (AGE) formation. We previously showed that AGEs produce an increase in vascular permeability and generated an oxidant stress after binding to the receptor (RAGE) present on endothelium. RAGE, a 35-kDa protein that belongs to the immunoglobulin superfamily, has been cloned from a rat lung cDNA library, and recombinant rat soluble RAGE (rR-RAGE) has been produced in insect cells. The sequence of RAGE is highly conserved between human and rat. We studied the biological effect of rR-RAGE and pharmacokinetics of 125I-rR-RAGE after intravenous or intraperitoneal administration in normal and streptozotocin induced diabetic rats. rR-RAGE prevented albumin or inulin transfer through a bovine aortic endothelial cell monolayer, restored the hyperpermeability observed in diabetic rats or induced in normal rats by diabetic rat red blood cells, and corrected the reactive oxygen intermediate production after intravenous or intraperitoneal administration. After intravenous injection of 125I-rR-RAGE, the distribution half-life was longer (p < or = 0.01) in diabetic (0.15 and 4.01 hr) than in normal (0.02 and 0.21 hr) rats, as was the case for the elimination half lives (diabetic, 57.17 hr; normal, 26.02 hr; p < or = 0.01). Distribution volume was higher in diabetic than in normal rats (6.94 and 3.24 liter/kg, respectively; p = 0.049). Our study showed that rR-RAGE was biologically active in vivo and slowly cleared, which suggests it could be considered as a potential therapy. PMID- 9224813 TI - Intracellular metabolism and action of acyclic nucleoside phosphonates on DNA replication. AB - 9-(2-phosphonylmethoxyethyl)guanine (PMEG) is an acyclic nucleoside phosphonate derivative that has demonstrated significant anticancer activity in a number of in vitro and in vivo animal model systems. In this study, we compared the cellular metabolism of PMEG and 9-(2-phosphonylmethoxyethyl)adenine (PMEA), a clinically active anti-HIV and antihepatitis agent, and the inhibitory activities of their putative active diphosphate derivatives, PMEGpp and PMEApp, respectively, toward human cellular DNA polymerases. PMEG was significantly more cytotoxic than PMEA against a panel of human leukemic cells. The diphosphate derivatives were the major metabolites formed in cells on both these agents, with PMEGpp reaching cellular concentration approximately 4-fold higher than that achieved for PMEApp. These differences in cellular accumulation of the diphosphate derivatives were not, however, sufficient to account for the 30-fold difference in cytotoxicity between the two analogs. PMEGpp was also at least a 7 fold more effective inhibitor of in vitro simian vacuolating virus 40 DNA replication system than that of PMEApp (IC50 = 4.6 microM). Studies with a defined primed DNA template showed that PMEGpp was a potent inhibitor of both human polymerases alpha and delta, two key enzymes involved in cellular DNA replication, whereas PMEApp inhibited these enzymes relatively poorly. From these studies, we can conclude that the factors that contribute to the enhanced antileukemic activity of PMEG derives both from its increased anabolic phosphorylation and the increased potency of the diphosphate derivative to target the cellular replicative DNA polymerases. PMID- 9224814 TI - Structural basis for differential induction of spermidine/spermine N1 acetyltransferase activity by novel spermine analogs. AB - The spermine analog N1,N11-diethylnorspermine (DE-333, also known as DENSPM or BENSPM) is regarded as the most potent known inducer of the polyamine catabolic enzyme, spermidine/spermine N1-acetyltransferase (SSAT), increasing activity by more than 200- to 1000-fold in certain cell types. The relative ability of a series of eight systematically modified DE-333 analogs to affect SSAT expression was examined in Malme-3M human melanoma cells, one of several cell lines known to be especially responsive to induction of this enzyme. In particular, we examined the relative contribution of induction of enzyme mRNA and prolongation of enzyme half-life to analog-mediated increases in enzyme activity. Induction of enzyme mRNA was most influenced by intra-amine carbon distances; relative effectiveness was found to be proportional to the number of three-carbon units. Stabilization of enzyme was most determined by the terminal N-alkyl substituent size; among methyl, ethyl and propyl groups, methyl was least effective. Thus, DE-333, which most potently induces SSAT mRNA and effectively stabilizes SSAT enzyme activity, produces the greatest increase in enzyme activity. Although other contributing mechanisms may be involved, the relative abilities of the various analogs to induce enzyme activity is at least partially attributable to their combined effects on enzyme mRNA and protein half-life. These data reveal the highly sensitive structure-activity relationships that underlie and control spermine analog induction of SSAT activity. Pending further definition of the relationship between SSAT induction and antitumor growth and toxicity in vivo, these relationships may be used to optimize therapeutic efficacy. PMID- 9224815 TI - Structure/activity relationships in lysophosphatidic acid: the 2-hydroxyl moiety. AB - Although lipid phosphoric acid mediators such as lysophosphatidic acid (LPA) are now recognized widely as intercellular signaling molecules, the medicinal chemistry of these mediators is poorly developed. With the goal of achieving a better understanding of the structure activity relationships in LPA, we have synthesized and tested a series of LPA analogs that lack the 2-hydroxyl moiety. Our series consisted of compounds with 2, 3, or 4 carbon diol or amino alcohol backbones and oleoyl or palmitoleoyl acyl groups. These molecules cannot be acylated further to form phosphatidic acids, nor do they have chiral centers. The rank order potency of these compounds in mobilization of calcium in MDA MB-231 cells suggested a maximum optimal chain length of 24-25 atoms. However, high potency for the inhibition of adenylyl cyclase in these cells was achieved only by one compound that also contained a dissociable proton five bond lengths from the phosphorus atom. That compound, N-oleoyl-2-hydroxyethyl-1-phosphate, was nearly equipotent to 1-oleoyl LPA in both assays. The striking mimicry of LPA by the ethanolamine-based compound and the presence of fatty acid amides in tissue prompts us to propose that phosphorylated N-acyl ethanolamides occur naturally. PMID- 9224816 TI - Novel 7-alkyl methylenedioxy-camptothecin derivatives exhibit increased cytotoxicity and induce persistent cleavable complexes both with purified mammalian topoisomerase I and in human colon carcinoma SW620 cells. AB - An alkylating camptothecin (CPT) derivative, 7-chloromethyl-10,11-methylenedioxy camptothecin (7-CM-MDO-CPT) was recently shown to produce irreversible topoisomerase I (top1) cleavage complexes by binding to the +1 base of the scissile strand of a top1 cleavage site. We demonstrate that 7-CM-EDO-CPT (7 chloromethyl-10,11-ethylenedioxy-camptothecin) also induces irreversible top1-DNA complexes. 7-CM-MDO-CPT, 7-CM-EDO-CPT, and the nonalkylating derivative 7-ethyl 10,11-methylenedioxy-camptothecin (7-E-MDO-CPT) also induced reversible top1 cleavable complexes, which were markedly more stable to salt-induced reversal than those induced by 7-ethyl-10-hyroxy-CPT, the active metabolite of CPT-11. This greater stability of the top1 cleavable complexes was contributed by the 7 alkyl and the 10,11-methylene- (or ethylene-) dioxy substitutions. Studies in SW620 cells showed that 7-E-MDO-CPT, 7-CM-MDO-CPT, and 7-CM-EDO-CPT are more potent inducers of cleavable complexes and more cytotoxic than CPT. The reversal of the cleavable complexes induced by 7-E-MDO-CPT, 7-CM-MDO-CPT, and 7-CM-EDO-CPT was markedly slower after drug removal than that for CPT, which is consistent with the data with purified top1. By contrast to CPT, 7-E-MDO-CPT, 7-CM-MDO-CPT, and 7-CM-EDO-CPT were cytotoxic irrespective of the presence of 10 microM aphidicolin. These results suggest that 7-E-MDO-CPT, 7-CM-MDO-CPT, and 7-CM-EDO CPT are more potent top1 poisons than CPT and produce long lasting top1 cleavable complexes and greater cytotoxicity than CPT in cells. PMID- 9224817 TI - Stimulation of protein kinase C rapidly reduces intracellular Na+ concentration via activation of the Na+ pump in OK cells. AB - Na+ reabsorption is regulated in proximal tubules by hormones that stimulate protein kinase C (PKC). To determine whether stimulation of PKC causes a reduction in intracellular Na+ concentration ([Na+]i) that might link Na+ pump activation to increased Na+ reabsorption, [Na+]i was measured in kidney cells loaded with the Na+-sensitive fluorescent indicator SBFI. Rapid digital imaging fluorescence microscopy determinations were performed in epithelial kidney cells transfected with the rodent Na+ pump alpha1 cDNA. In 42 determinations, the basal [Na+]i was 19.7 +/- 2.4 mM. Stimulation of PKC reduced the [Na+]i to 5.6 +/- 0.6 mM in approximately 10 sec. This drastic change in [Na+]i requires a transient 74 120-fold increase in Na+ pump activity. After the new steady state [Na+]i is reached, the Na+ pump is 58% activated. The entry of Na+ into the cells is not affected by stimulation of PKC; therefore, the reduction in [Na+]i is exclusively dependent on activation of the Na+ pump. Accordingly, PKC stimulation does not affect the [Na+]i of cells expressing a mutant Na+ pump that is not stimulated by PKC. The decrease in [Na+]i observed in cells transfected with the rodent Na+ pump alpha1 cDNA is large and sufficiently fast that it is expected to stimulate rapidly passive Na+-influx into the cells, thereby accounting for the observed PKC-induced stimulation of Na+ reabsorption. PMID- 9224818 TI - Development of resistance of human immunodeficiency virus type 1 to dextran sulfate associated with the emergence of specific mutations in the envelope gp120 glycoprotein. AB - Polyanionic compounds are known to inhibit the binding of human immunodeficiency virus (HIV) to CD4+ cells and the subsequent fusion step between the virus and cells. We selected an HIV-1 strain resistant to dextran sulfate (DS) by cultivation of HIV-1 (NL4-3)-infected MT-4 cells in the presence of DS Mr 5000. DS did not inhibit the binding of DS-resistant virus to MT-4 cells or syncytium formation between MOLT cells and HUT-78 cells persistently infected with the DS resistant virus. In addition, a monoclonal antibody with specificity for the V3 loop of envelope gp120 glycoprotein did not recognize the DS-resistant HIV-1 gp120 V3 loop. The following mutations were found in the gp120 molecule of the DS resistant HIV-1 strain but not in the wild-type strain: S114N in the V1 loop region; S134N in the V2 loop region; K269E, Q278H, and N293D in the V3 loop region; N323S in the C3 region; a deletion of five amino acids (Phe-Asn-Ser-Thr Trp) at positions 364-368 in the V4 loop; and R3871 in the CD4 binding domain. Our results suggest that (i) DS interacts with specific amino acid residues in the gp120 molecule, (ii) the virus is able to overcome the inhibitory effect of DS on viral infectivity, (iii) cross-resistance developed against those polyanionic compounds that are structurally related to DS, and (iv) the molecular determinants of HIV cell tropism, syncytium-inducing ability, coreceptor (fusin/ CC-CKR5) utilization, and polyanion resistance seem to be located in the env genome of HIV and specifically in the V3 loop domain. PMID- 9224819 TI - The mu-opioid receptor down-regulates differently from the delta-opioid receptor: requirement of a high affinity receptor/G protein complex formation. AB - Chronic opioid treatment of Neuro2A cells stably expressing either delta-opioid receptor (DOR) or mu-opioid receptor (MOR) resulted in agonist-dependent receptor down-regulation. Although there is high homology in the DOR and MOR amino acid sequences, there is an apparent difference in the regulation of the cellular levels of these two receptors. The ability of 24-hr [D-Pen2,D-Pen5]enkephalin (DPDPE) treatment to internalize and down-regulate DORs expressed in Neuro2A remained intact after pertussis toxin (PTX) pretreatment, which uncouples the receptor from G proteins. In contrast, the ability of [D-Ala2,N-MePhe4,Gly ol5]enkephalin (DAMGO) to internalize and down-regulate MORs in Neuro2A cells was completely abolished by PTX pretreatment. The requirement of functional MOR but not DOR in agonist-induced receptor down-regulation was further demonstrated by site-directed mutagenesis of the receptors. When Asp114 in transmembrane 2 of MOR was converted to alanine, the ability was abolished of DAMGO or morphine to inhibit forskolin-stimulated [3H]cAMP production in Neuro2A cells stably expressing this mutant receptor. There was a parallel decrease in agonist affinity and elimination of the agonist-induced receptor down-regulation. On the other hand, although the equivalent mutation of Asp95 to alanine in DOR likewise resulted in the inability of DPDPE to inhibit [3H]cAMP production, the ability of DPDPE to down-regulate this mutant receptor after 24-hr treatment was unaffected. This difference in MOR and DOR down-regulation could be caused by the differences in the ability of these two receptors to form high affinity complexes with G proteins. DOR retained the ability to form high affinity complexes even after PTX pretreatment or after mutation of Asp95 in transmembrane 2. In contrast, MOR existed only in the low affinity, uncoupled state after PTX pretreatment or after conversion of Asp114 to alanine. Therefore, in Neuro2A cells, agonist-induced opioid receptor down-regulation seems to depend directly on the formation of the high affinity receptor complexes and not on the activation of the receptors and subsequent transduction of the signals. PMID- 9224820 TI - Lactone modulation of the gamma-aminobutyric acid A receptor: evidence for a positive modulatory site. AB - The gamma-aminobutyric acid-A (GABA(A)) receptor complex is allosterically modulated by a variety of substances, some of clinical importance. Barbiturates and neurosteroids augment GABA-currents and also directly gate the channel. A variety of gamma-butyrolactone analogues also modulate GABA-induced currents, with some potentiating and others inhibiting. Because several gamma thiobutyrolactone analogues have biphasic effects on GABA currents, experiments with wild-type and picrotoxinin-insensitive GABA(A) receptors were performed to analyze whether some gamma-thiobutyrolactones interact with two distinguishable sites on the GABA(A) receptor. beta-Ethyl-beta-methyl-gamma-thiobutyrolactone inhibited GABA-induced currents at low concentrations (0.001-1 mM), but potentiated GABA-induced currents at higher concentrations (3-10 mM) in wild-type alpha1beta2gamma2-subunit containing ionophores. The related alpha-ethyl-alpha methyl-gamma-thiobutyrolactone potentiated submaximal GABA currents in wild-type receptors at both low and high concentrations (0.1-10 mM). Mutations in the second transmembrane domain of alpha1, beta2, or gamma2 conferred picrotoxinin insensitivity onto GABA(A) receptor complexes. When these mutated alpha1, beta2, or gamma2 subunits were incorporated into the receptor complex, beta-ethyl-beta methyl-gamma-thiobutyrolactone potentiated GABA currents over the entire concentration range (0.1-10 mM). Neither the potentiating activity nor the EC50 of alpha-ethyl-alpha-methyl-gamma-thiobutyrolactone changed in the mutant receptors. Further studies demonstrated that the mutations did not affect the EC50 of chlordiazepoxide or phenobarbital. These and our earlier results identify a modulatory site on the GABA(A) receptor distinct from that interacting with barbiturates, benzodiazepines, and steroids. Additionally, they show that the gamma-butyrolactones probably interact at two different sites on the ionophore to produce opposite effects on GABA-mediated current. PMID- 9224821 TI - High affinity binding of [3H]propionyl-[Met(O2)11]substance P(7-11), a tritiated septide-like peptide, in Chinese hamster ovary cells expressing human neurokinin 1 receptors and in rat submandibular glands. AB - Propionyl-[Met(O2)11]substance P(7-11) [ALIE-124 or propionyl-[Met(O2)11]SP(7 11)] has been designed as a septide-like ligand adequate for tritiation and, therefore, adequate for binding studies. In Chinese hamster ovary (CHO) cells expressing human tachykinin neurokinin (NK)-1 receptors, ALIE-124 displaced [3H][Pro9]substance P (SP) from its binding site at micromolar concentrations. However, ALIE-124 stimulated phosphatidylinositol hydrolysis, as previously shown for septide-like peptides. With [3H]ALIE-124 (95 Ci/mmol), we have been able to reveal a high affinity binding site in CHO cells (Kd = 6.6 +/- 1.0 nM), with a low maximal binding capacity. [3H]ALIE-124 specific maximal binding represented only 15-20% of that observed with [3H][Pro9]SP in CHO cells. Septide-like peptides, including septide and NKA, were potent competitors (in the nanomolar range) of [3H]ALIE-124 specific binding site. Interestingly, SP and [Pro9]SP were also potent competitors, with 10-fold greater potency for sites labeled with [3H]ALIE-124 than for sites labeled with [3H][Pro9]SP. The NK-1 antagonist RP 67580 also showed a higher potency for [3H]ALIE-124 than for [3H][Pro9]SP specific binding sites. NKB and [Lys5,methyl-Leu9,Nle10]NKA(4-10) displaced [3H]ALIE-124 binding but with lower potency, whereas senktide had no affinity. The existence of [3H]ALIE-124 specific binding sites was also demonstrated in rat submandibular gland. In this tissue, [3H]ALIE-124 specific maximal binding was higher, reaching 40-50% of that achieved with [3H][Pro9]SP. PMID- 9224822 TI - Vasoactive intestinal polypeptide and pituitary adenylate cyclase-activating polypeptide receptor chimeras reveal domains that determine specificity of vasoactive intestinal polypeptide binding and activation. AB - Vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase activating polypeptide (PACAP) receptors are closely related G protein-coupled receptors with seven-transmembrane domains. The VIP receptor can bind both VIP and PACAP with high affinity, whereas the PACAP receptor binds only PACAP with high affinity. To elucidate the structural domains involved in a selectivity for VIP binding and the subsequent receptor activation, a series of chimeric receptors between the VIP and PACAP receptors was constructed, expressed in COS-7 cells, and analyzed for ligand binding and cAMP generation. All chimeric constructs bound PACAP with high affinity and subsequently activated cAMP generation similarly to the wild-type receptors. In contrast, profound differences were observed in the potencies of VIP for competition of 125I-labeled PACAP binding to both wild-type receptors and the chimeric receptors. The cAMP responses of these receptors generally correlated with the ability of VIP to compete for PACAP radioligand binding with the exceptions for some particular chimeras. In this report we demonstrate that several domains, including the amino terminal extracellular domain, the transmembrane domains I and II, and the first extracellular loop of the VIP receptor, are important for the selectivity for VIP binding and responsiveness to VIP. We further show that the third extracellular loop and its proximal domains of the VIP receptor appear to be involved in the VIP recognition, especially the receptor activation process. On the other hand, the direct binding experiments of the VIP radioligand demonstrated that both wild type receptors and all chimeric receptors have a high affinity binding site for VIP, although this high affinity VIP binding resulted in a biological response only in the VIP receptor or VIP receptor-like chimeras. This suggests that there is a nonbiologically relevant high affinity VIP-binding site within the rat PACAP receptor. PMID- 9224823 TI - Efficacy and kinetics of opioid action on acutely dissociated neurons. AB - Opioids have been shown to cause a potent inhibition of neurons in the locus ceruleus (LC) in vivo in brain slices and isolated neurons; however, the kinetics of opioid action have not been described. In this study, we used acutely isolated LC neurons to examine opioid and alpha2-adrenoceptor action on potassium and calcium currents. [Met]Enkephalin (ME), [D-Ser2,Leu5,Thr6]-enkephalin, etorphine, and [D-Ala2,N-Me-Phe4,Gly-ol5]enkephalin increased potassium conductance, whereas morphine and naloxone were antagonists. The time constant of potassium channel activation was approximately 0.7 sec and was the same for each agonist. The amplitude of the current and the time constant of decay were dependent on the agonist, suggesting that agonist efficacy and affinity, respectively, determined these parameters. The amplitude of potassium current induced by the alpha2 adrenoceptor agonist UK14304 was not significantly different from that induced by ME, but the time constant of current activation was half that of ME, and the decline was more rapid. When potassium conductances were blocked with the combination of internal cesium and external barium, opioid and alpha2 agonists had no effect at potentials more negative than -50 mV and decreased barium currents at potentials between -40 and +20 mV. Both morphine and clonidine caused a small inhibition of barium current. In dorsal root ganglion cells, morphine alone had small and inconsistent effects on the calcium current, but it always competitively antagonized the inhibition caused by [D-Ala2,N-Me-Phe4,Gly ol5]enkephalin. The results in isolated LC neurons suggest 1) the amplitude and time course of the opioid-induced potassium current depend on agonist efficacy and affinity and 2) the coupling of both mu-opioid and alpha2-adrenoceptors to calcium channels seems to be more efficient than that to potassium channels. PMID- 9224824 TI - The stability of the agonist beta2-adrenergic receptor-Gs complex: evidence for agonist-specific states. AB - A restricted version of the ternary complex model for receptor-G protein complex formation has recently been proposed. Known as the two-state model, this model proposes that in the context of agonist and G protein interactions, only two thermodynamic states exist for the receptor: active (R*) and inactive (R). One form of this model suggests that only the R* state of the receptor is capable of interacting with and subsequently activating G proteins. We directly tested the kinetic aspects of a strict two-state receptor model in a cell line containing the native beta2-adrenergic receptor that is capable of inducing Gs expression. We examined adenylyl cyclase activity in the presence of limiting GTP levels and concluded that there exists a different rate of heterotrimer dissociation (i.e., HR*G yields HR* + G*) for different beta2-agonists. This finding is inconsistent with a strict two-state model in which R* is a characteristic of the receptor that is independent of the identity of the agonist. It implies that agonist activation of adenylyl cyclase is more complicated than a simple two-state model. PMID- 9224826 TI - A conserved threonine residue in the second intracellular loop of the 5 hydroxytryptamine 1A receptor directs signaling specificity. AB - Productive interaction between receptors and G proteins involves multiple intracellular receptor domains, but the role of individual receptor amino acids in directing the selection of specific signaling pathways has not yet been identified. Sequence alignment of several G protein-coupled receptors identified a highly conserved threonine residue in the i2 loop of the 5-hydroxytryptamine 1A (5-HT1A) receptor that is a putative protein kinase C phosphorylation consensus site and is located in a predicted amphipathic alpha-helical domain. To examine the role of this conserved threonine residue in 5-HT1A receptor coupling to Gi/Go proteins, this residue was mutated to alanine (T149A mutant). Wild-type and mutant 5-HT1A receptors were stably transfected into both Ltk- and GH4C1 cells to investigate receptor coupling to multiple signaling pathways. In both cell lines, the T149A mutant displayed similar agonist affinities as the wild-type receptor. In Ltk- cells, the T149A 5-HT1A receptor inhibited cAMP accumulation by 30% compared with wild-type (83%). A 2.6-fold increase in intracellular calcium (due to phospholipase C-mediated calcium mobilization) was observed for the wild-type receptor upon the addition of 100 nM 5-HT; whereas the T149A 5-HT1A receptor failed to mediate a calcium mobilization response at equivalent receptor levels to wild-type. When transfected in GH4C1 cells, the T149A receptor mutant fully inhibited basal cAMP and partially inhibited Gs-stimulated cAMP accumulation compared with wild-type receptor (57 +/- 14% versus 86 +/- 2%). In contrast, the T149A 5-HT1A receptor mutant failed to block the influx of calcium induced by calcium channel agonist (+/-)-Bay K8644, whereas the wild-type 5-HT1A receptor inhibited the calcium influx by 40%. Thus, the Thr149 residue is directly involved in G protein coupling to calcium mobilization (mediated by betagamma subunits of Gi2) and to inhibition of calcium channel activation (mediated by betagamma subunits of Go) but plays a minor role in coupling to alpha1-mediated inhibition of cAMP accumulation. The conserved i2 loop threonine may serve as a G protein contact site to direct the signaling specificity of multiple receptors. PMID- 9224825 TI - Differences in folylpolyglutamate synthetase and dihydrofolate reductase expression in human B-lineage versus T-lineage leukemic lymphoblasts: mechanisms for lineage differences in methotrexate polyglutamylation and cytotoxicity. AB - Cellular accumulation of methotrexate polyglutamates (MTXPGs) is recognized as an important determinant of the cytotoxicity and selectivity of methotrexate in acute lymphoblastic leukemia (ALL). We identified a significantly lower cellular accumulation of MTXPGs in T-lineage versus B-lineage lymphoblasts in children with ALL, which is consistent with the worse prognosis of T-lineage ALL when treated with conventional antimetabolite-based therapy. Maximum MTXPG accumulation in leukemic blasts in vivo was 3-fold greater in lymphoblasts of children with B-lineage ALL (129 children) compared with those with T-lineage ALL (20 children) (p < 0.01) and was characterized by a saturable (Emax) model in both groups. The human leukemia cell lines NALM6 (B-lineage) and CCRF/CEM (T lineage) were used to assess potential mechanisms for these lineage differences in MTX accumulation, revealing i) greater total and long-chain MTXPG accumulation in NALM6 over a wide range of methotrexate concentrations (0.2-100 microM), ii) saturation of MTXPG accumulation in both cell lines, with a higher maximum (Emax in NALM6, iii) 3-fold higher constitutive FPGS mRNA expression and enzyme activity in NALM6 cells, iv) 2-fold lower levels of DHFR mRNA and protein in NALM6 cells, and v) 4-6 fold lower extracellular MTX concentration and 2-fold lower intracellular MTXPG concentration to produce equivalent cytotoxicity (LC50) in NALM6 versus CEM. There was a significant relationship between FPGS mRNA and enzyme activity in lymphoblasts from children with newly diagnosed ALL, and blast FPGS mRNA and activity increased after methotrexate treatment. These data indicate higher FPGS and lower DHFR levels as potential mechanisms contributing to greater MTXPG accumulation and cytotoxicity in B-lineage lymphoblasts. PMID- 9224828 TI - Differential roles of monkey striatum in learning of sequential hand movement. AB - To study the role of the basal ganglia in learning of sequential movements, we trained two monkeys to perform a sequential button-press task (2x5 task). This task enabled us to examine the process of learning new sequences as well as the execution of well-learned sequences repeatedly. We injected muscimol (a GABA agonist) into different parts of the striatum to inactivate the local neural activity reversibly. The learning of new sequences became deficient after injections in the anterior caudate and putamen, but not the middle-posterior putamen. The execution of well-learned sequences was disrupted after injections in the middle-posterior putamen and, less severely, after injections in the anterior caudate/putamen. These results suggest that the anterior and posterior portions of the striatum participate in different aspects of learning of sequential movements. PMID- 9224827 TI - Positive cooperativity of acetylcholine and other agonists with allosteric ligands on muscarinic acetylcholine receptors. AB - It is well known that allosteric modulators of muscarinic acetylcholine receptors can both diminish and increase the affinity of receptors for their antagonists. We investigated whether the allosteric modulators can also increase the affinity of receptors for their agonists. Twelve agonists and five allosteric modulators were tested in experiments on membranes of CHO cells that had been stably transfected with genes for the M1-M4 receptor subtypes. Allosterically induced changes in the affinities for agonists were computed from changes in the ability of a fixed concentration of each agonist to compete with [3H]N-methylscopolamine for the binding to the receptors in the absence and the presence of varying concentrations of allosteric modulators. The effects of allosteric modulators varied greatly depending on the agonists and the subtypes of receptors. The affinity for acetylcholine was augmented by (-)-eburnamonine on the M2 and M4 receptors and by brucine on the M1 and M3 receptors. Brucine also enhanced the affinities for carbachol, bethanechol, furmethide, methylfurmethide, pilocarpine, 3-(3-pentylthio-1,2,5-thiadiazol-4-yl)-1,2,5,6-tetrahydro-1- methylpyridine (pentylthio-TZTP), oxotremorine-M, and McN-A-343 on the M1, M3, and M4 receptors, for pentylthio-TZTP on the M2 receptors, and for arecoline on the M3 receptors. ( )-Eburnamonine enhanced the affinities for carbachol, bethanechol, furmethide, methylfurmethide, pentylthio-TZTP, pilocarpine, oxotremorine and oxotremorine-M on the M2 receptors and for pilocarpine on the M4 receptors. Vincamine, strychnine, and alcuronium displayed fewer positive allosteric interactions with the agonists, but each allosteric modulator displayed positive cooperativity with at least one agonist on at least one muscarinic receptor subtype. The highest degrees of positive cooperativity were observed between (-)-eburnamonine and pilocarpine and (-)-eburnamonine and oxotremorine-M on the M2 receptors (25- and 7-fold increases in affinity, respectively) and between brucine and pentylthio TZTP on the M2 and brucine and carbachol on the M1 receptors (8-fold increases in affinity). The discovery that it is possible to increase the affinity of muscarinic receptors for their agonists by allosteric modulators offers a new way to subtype-specific pharmacological enhancement of transmission at cholinergic (muscarinic) synapses. PMID- 9224829 TI - Identification of motor and sensory brain activities during unilateral finger movement: spatiotemporal source analysis of movement-associated magnetic fields. AB - We investigated the movement-related cortical fields (MRCFs) recorded by magnetoencephalography (MEG) to identify the motor and sensory brain activities at the instant of the unilateral finger movement using six normal subjects. We focused our investigation on the source analysis of the events tightly linked to movement onset, and we used brain electric source analysis (BESA) to model the sources generating MRCFs during the interval from 200 ms before to 150 ms after the movement onset. Four sources provided satisfactory solutions for MRCF activities in this interval. Sources 1 and 2, which were located in the pre central regions in the hemisphere contralateral and ipsilateral to the moved finger, respectively, generated the readiness fields (RF), but source 1 was predominant just before movement onset. The motor field (MF), the peak of which was just after movement onset, was mainly generated by source 1. Sources 3 and 4 were located in the post-central regions in the hemisphere contralateral and ipsilateral to the moved finger, respectively. The first motor evoked field (MEF I), the peak of which was about 80 ms after the movement, was mainly generated by source 3, but with the participation of sources 1, 2 and 4. The results indicated that the activities of both pre -and post-central regions in bilateral hemispheres were related to voluntary movements, although the predominant areas varied over time. This is the first noninvasive study to clarify the complex spatiotemporal activities relating movements in humans using a multi-channel MEG system. PMID- 9224830 TI - Inhibitors of G-proteins and protein kinases reduce the sensitization to mechanical stimulation and the desensitization to heat of spinothalamic tract neurons induced by intradermal injection of capsaicin in the primate. AB - Intradermal injection of capsaicin results in sensitization of spinothalamic tract cells to brushing and pressure applied to the cutaneous receptive field in anesthetized monkeys. A significant increase in background activity also occurs immediately after capsaicin injection that lasts for at least 2 h. A 40-50% decrease in the response to noxious heat stimuli is also observed following capsaicin injection. This study investigated the spinal role of second messengers by extracellularly recording from spinothalamic tract cells and delivering inhibitors of second messenger pathways to the spinal cord by microdialysis. Blockade of protein kinases with the general protein kinase inhibitor, H7 (5.0 mM, n = 6), reduced the sensitization of the cells to brush and pressure. Blockade of protein kinase C with NPC15437 (10.0 mM, n = 10) reduced the increased background activity and the increased responses to brush. Blockade of protein kinase A with H89 (0.01 mM, n = 9) was most effective. H89 reduced the background activity, the increased responses to brush and press, and reversed the decreased response to noxious heat stimuli. Blockade of G-proteins with the general G-protein inhibitor, GDP-beta-S (1.0 mM, n = 9), reduced the background activity and the responses to brush and pressure without affecting the decreased response to heat. Thus, multiple intracellular messengers appear to be involved in the processing of central sensitization induced by activation of C-fibers following intradermal injection of capsaicin. PMID- 9224831 TI - Saccadic eye movements to visual and auditory targets. AB - Recent neurophysiological studies of the saccadic ocular motor system have lent support to the hypothesis that this system uses a motor error signal in retinotopic coordinates to direct saccades to both visual and auditory targets. With visual targets, the coordinates of the sensory and motor error signals will be identical unless the eyes move between the time of target presentation and the time of saccade onset. However, targets from other modalities must undergo different sensory-motor transformations to access the same motor error map. Because auditory targets are initially localized in head-centered coordinates, analyzing the metrics of saccades from different starting positions allows a determination of whether the coordinates of the motor signals are those of the sensory system. We studied six human subjects who made saccades to visual or auditory targets from a central fixation point or from one at 10 degrees to the right or left of the midline of the head. Although the latencies of saccades to visual targets increased as stimulus eccentricity increased, the latencies of saccades to auditory targets decreased as stimulus eccentricity increased. The longest auditory latencies were for the smallest values of motor error (the difference between target position and fixation eye position) or desired saccade size, regardless of the position of the auditory target relative to the head or the amplitude of the executed saccade. Similarly, differences in initial eye position did not affect the accuracy of saccades of the same desired size. When saccadic error was plotted as a function of motor error, the curves obtained at the different fixation positions overlapped completely. Thus, saccadic programs in the central nervous system compensated for eye position regardless of the modality of the saccade target, supporting the hypothesis that the saccadic ocular motor system uses motor error signals to direct saccades to auditory targets. PMID- 9224832 TI - Further evidence for non-monosynaptic group I excitation of motoneurones in the human lower limb. AB - Non-monosynaptic group I and group II excitation of human lower limb motoneurones was investigated. Changes in the firing probability of individual voluntarily activated motor units belonging to various muscles (soleus, gastrocnemius medialis, tibialis anterior, peroneus brevis, quadriceps and biceps femoris) were investigated after stimulation of various nerves (posterior tibial, common peroneal and femoral nerves) with weak (0.4-0.6x motor threshold) electrical stimuli. In all investigated motor nuclei, stimulation of the "homonymous" nerve evoked a peak of increased firing probability with a latency that was 3-7 ms longer than the monosynaptic Ia latency. The more caudal the motor nucleus explored, the greater the central delay. This strongly suggests a transmission through neurones located above the lumbar enlargement. If one excepts the sural induced excitation of peroneus brevis units, which seems to be mediated through a particular pathway, the main peripheral input to neurones mediating non monosynaptic excitation evoked by these weak stimuli is group I in origin. The pattern of distribution of non-monosynaptic group I excitation was very diffuse, since stimulation of each nerve was able to evoke excitation in all investigated nuclei. In most cases, non-monosynaptic excitation evoked in a given motor unit by stimulation of one nerve was depressed on combined stimulation of two nerves, and evidence is presented that this lateral inhibition is exerted at a premotoneuronal level. By contrast, there was no evidence that increasing the afferent input in a given pathway evokes an "autogenetic" inhibition in this pathway. The negative correlation found between non-monosynaptic group I-induced and late group II-induced facilitation of the quadriceps H-reflex when using high stimulus intensities applied on the common peroneal nerve suggests that these two effects could be mediated through common interneurones. PMID- 9224833 TI - Intrinsic circuitry and physiological properties of pyramidal neurons in rat barrel cortex. AB - Pyramidal neurons in the rat posteromedial barrel subfield (PMBSF) were characterized physiologically and filled with biocytin in in vitro brain slices. Intrinsic axons belonging to supragranular neurons projected horizontally and vertically, arborizing in layers II/III and V, but had few or no projections to layers IV or VI. These axons projected horizontally for up to 2 mm, spanning two to seven barrel columns. Layer V neurons had more diffuse axon arbors that projected either vertically, arborizing in layers III to V, or horizontally, branching profusely in layers V and VI. The basal dendritic trees of neurons in layers II/III, V and VI spanned one or two barrel columns without being skewed toward particular barrel columns. Physiologically, regular-spiking neurons were classified as "RS1" or "RS2" according to their degree of late spike frequency adaptation. RS1 neurons were found in superficial and deep layers, whereas RS2 neurons were significantly more prevalent in the latter. Infragranular, but not supragranular neurons showed slow, inward rectification at hyperpolarized potentials. All neurons generated fast and medium afterhyperpolarizations following individual spikes; however, only infragranular pyramids had depolarizing afterpotentials interposed between the two afterhyperpolarizations. RS1 neurons had larger cell bodies, longer total basal dendritic lengths, and more densely branched proximal dendritic trees than RS2 neurons. These findings indicate that pyramidal neurons in the deep and superficial layers of the rat PMBSF have distinct patterns of intracortical axon arbors and distinct physiological properties. These features are probably involved in shaping and modulating the response properties of PMBSF neurons. PMID- 9224834 TI - Smooth-pursuit eye movements elicited by first-order and second-order motion. AB - The perception of the displacement of luminance-defined contours (i.e., first order motion) is an important and well-examined function of the visual system. It can be explained, for example, by the operation of elementary motion detectors (EMDs), which cross-correlate the spatiotemporal luminance distribution. More recent studies using second-order motion stimuli, i.e., shifts of the distribution of features such as contrast, texture, flicker, or motion, extended classic concepts of motion perception by including nonlinear or hierarchical processing in the EMD. Smooth-pursuit eye movements can be used as a direct behavioral probe for motion processing. The ability of the visual system to extract motion signals from the spatiotemporal changes of the retinal image can be addressed by analyzing the elicited eye movements. We measured the eye movement response to moving objects defined by two different types of first-order motion and two different types of second-order motion. Our results clearly showed that the direction of smooth-pursuit eye movements was always determined by the direction of object motion. In particular, in the case of second-order motion stimuli, smooth-pursuit did not follow the retinal image motion. The latency of the initial saccades during pursuit of second-order stimuli was slightly but significantly increased, compared with the latency of saccades elicited by first order motion. The processing of second-order motion in the peripheral visual field was less exact than the processing of first-order motion in the peripheral field. Steady state smooth-pursuit eye speed did not reflect the velocity of second-order motion as precisely as that of first-order motion, and the resulting retinal error was compensated by saccades. Interestingly, for slow second-order stimuli we observed that the eye could move faster than the target, leading to small, corrective saccades in the opposite direction to the ongoing smooth pursuit eye movement. We conclude from our results that both visual perception and the control of smooth-pursuit eye movements have access to processing mechanisms extracting first- and second-order motion. PMID- 9224835 TI - Effects of graft pooling of foetal rat and mouse tissue and immunosuppression in the 6-hydroxydopamine rat model of Parkinson's disease. AB - We employed intracerebral co-transplantation of foetal xenogeneic striatal mouse tissue and allogeneic rat substantia nigra into the adult rat brain to elucidate the effects of xenogeneic mouse graft on the function and survival of an allogeneic rat graft in 6-hydroxydopamine lesioned Sprague-Dawley rats. Foetal mouse striatum (STR) and rat substantia nigra (VM) were transplanted as non pooled separate deposits or a pooled cell suspension with or without cyclosporin A (Cy A). Immunosuppressed recipients of pooled rat and mouse co-grafts showed a significantly better compensation of amphetamine-induced rotational behaviour compared with non-immunosuppressed animals with pooled rat and mouse co-grafts 3 and 6 weeks post-grafting. Tyrosine hydroxylase (TH) immunohistochemistry revealed a non-significant reduction in survival in pooled (1806.3+/-367.5 cells) rat and mouse co-transplants without immunosuppression compared with immunosuppressed pooled (3383.3+/-732.7 cells) animals with allo- and xenogeneic tissue and controls (3506.4+/-839.3 cells). Graft volumes were significantly reduced in pooled transplants without immunosuppression (0.1+/-0.026 mm3; ANOVA post-hoc Scheffe F-test, P<0.0001) compared with immunosuppressed recipients (0.7+/-0.1 mm3) and controls (0.6+/-0.1 mm3). In non-pooled allo- and xenogeneic grafts without immunosuppression the survival rate of the TH-immunoreactive cells and graft volumes were reduced (2359.3+/-479.5 cells; 0.2+/-0.043 mm3) compared with immunosuppressed animals (2927.3+/-946.6 cells; 0.6+/-0.2 mm3) and controls (2701.1+/-693.8 cells; 0.3+/-0.1 mm3) without reaching a level of significance. Rejection of mouse tissue was observed in all non-immunosuppressed recipients. In summary: (i) continued immunosuppression yielded significant beneficial effects on function and beneficial effects on survival of pooled grafts with an immunogenetic disparity; (ii) the rejection of a xenogeneic graft component may compromise survival and function of other, allogeneic graft components; and (iii) transplantation of non-pooled allo- and xenogeneic tissues may result in a better survival of the graft compared with pooled cell suspensions. PMID- 9224836 TI - Learning impairment induced by lesion of the CA1 field of the primate hippocampus: attempts to ameliorate the impairment by transplantation of fetal CA1 tissue. AB - Monkeys with bilateral excitotoxic lesion of the CA1 field of the hippocampus were severely impaired at learning visuospatial conditional tasks. This was not a general spatial impairment, because the animals were not impaired on serial spatial reversal, which requires response flexibility in the spatial domain; they were not impaired at learning to choose the position furthest away from a single stimulus, which requires analysis of spatial layout of the test area, and they were not impaired at discriminating between two patterns that differed only in orientation. CA1-lesioned monkeys were impaired at learning a visuospatial conditional task when trials of the two component types "if AA go left" and "if BB go right" were presented according to either a pseudorandom or alternating schedule; but they were not impaired if one component type of trial was presented until three consecutive correct responses were made, followed by the other type of trial, to three consecutive correct responses. In all cases testing continued until a criterion of 27 of 30 consecutive correct responses across both types of trial was achieved. Although this suggests that CA1-lesioned animals are particularly prone to interference effects, they had no difficulty in learning ten concurrent visual discriminations presented against either a uniform background or with each discrimination presented against its own distinctive background, a condition that might reduce interference in unoperated monkeys. Interference following hippocampal damage might occur at a deeper level than stimulus identification such that animals with hippocampal damage may be able to learn about many aspects of different stimuli in parallel but may be unable to learn about multiple related aspects of the same subject matter. Monkeys with grafts of fetal CA1 tissue in the lesioned CA1 field showed significant improvement relative to CA1-lesioned animals on those tasks on which CA1-lesioned animals were impaired, although they remained impaired relative to control animals. This suggests that the grafts had produced some improvement in performance. Grafted monkeys did not differ from unoperated control monkeys or from CA1-lesioned monkeys on those tasks that were not sensitive to CA1 damage. This demonstrates that the grafts did not have an additional deleterious effect on cognitive performance. PMID- 9224837 TI - Glutamate, aspartate and co-localization with calbindin in the medial thalamus. An immunohistochemical study in the rat. AB - Topographical and quantitative features of medial thalamic neurons in which aspartate (ASP) or glutamate (GLU) might act as neurotransmitters were investigated in the rat. The calcium-binding protein calbindin D-28k (CB) was exploited as a marker of neuronal subsets, thus allowing us to study also the relationships between the CB-containing neurons and those immunoreactive to excitatory amino acids. Double immunocytochemistry of ASP and CB or GLU and CB was performed in 40-microm-thick sections. The three markers were distributed in the thalamic midline, mediodorsal, anterior intralaminar and ventromedial nuclei, with regional variations. ASP-immunoreactive neurons appeared more numerous than the GLU-immunoreactive ones throughout these structures; ASP-CB or GLU-CB double immunostained neurons were evident. ASP-, GLU- and CB-immunoreactive cells were then quantitatively evaluated in 5-microm-thick consecutive sections. Interindividual variations and different anti-ASP and anti-GLU antibodies did not result in significant differences. ASP and GLU were not co-localized. Single ASP- or GLU-immunoreactive neurons accounted for 60% of the total number of immunostained cells, and single ASP-immunopositive cells represented more than half of these neurons. Among the CB-immunoreactive cells (40% of the total), half were double immunostained; the proportion of double CB-ASP-immunopositive neurons was sevenfold higher than that of the CB-GLU-immunoreactive ones. These results indicate that ASP may act as excitatory neurotransmitter in a relatively high proportion of medial thalamic neurons, in which ASP frequently coexists with CB. Approximately 50% of the CB-immunoreactive cells did not contain either ASP or GLU, suggesting that some medial thalamic neurons may utilize a different neurotransmitter. PMID- 9224838 TI - Delayed neuronal death following perinatal asphyxia in rat. AB - The consequences of perinatal asphyxia on the rat brain were studied 80 min to 8 days after birth with hematoxylin-eosin and in situ DNA double-strand-breaks labeling histochemistry. Asphyxia was induced by immersing fetus-containing uterus horns, removed from ready-to-deliver Sprague-Dawley rats, in a water bath at 37 degrees C for various time periods (0-22 min). Spontaneous- and cesarean delivered pups were used as controls. Perinatal asphyxia led to a decrease in the rate of survival, depending upon the length of the insult. No gross morphological changes could be seen in the brain of either control or asphyctic pups at any of the studied time points after delivery. However, in all groups, nuclear chromatin fragmentation, corresponding to in situ detection of DNA fragmentation, was observed at different stages. Nuclear fragmentation in control pups showed a specific distribution that appeared to be related to brain maturation, thus indicating programmed cell death. A progressive and delayed increase in nuclear fragmentation was found in asphyctic pups, which was dependent upon the length of the perinatal insult. The most evident effect was seen in frontal cortex, striatum, and cerebellum at postnatal day 8, although changes were also found in ventral-posterior thalamus, at days 1 and 2. Thus, nuclear chromatin fragmentation in asphyctic pups indicates a delayed post-asphyctic neuronal death. The absence of signs of inflammation or necrosis suggests that delayed neuronal cell death following perinatal asphyxia is an active, apoptosis-like phenomenon. PMID- 9224840 TI - Expression of Fos-like immunoreactivity in the brain and spinal cord of rats following middle cerebral artery occlusion. AB - This study examined c-fos protein expression in the brain and spinal cord of rats following permanent occlusion of the middle cerebral artery (MCA) above the rhinal fissure. At 1 h after right-sided MCA occlusion, Fos-like immunoreactivity (Fos-LI) was detected in neurons not only in the ipsilateral cerebral cortex but also in the spinal cord. In the latter, Fos-LI was localized in the nucleus and perikarya of neurons in the grey matter, notably the large motor neurons in the ventral horn. Fos-LI was most intense at 2-4 h, but became undetectable after 48 h in the cerebral cortex and 72 h in the spinal cord. In sham-operated animals, Fos-LI was almost undetectable or virtually absent. It was also not detected in the core territory supplied by the MCA at any time points after arterial occlusion. When the ischaemia-induced neuronal damage in both the cerebral cortex and spinal cord was evaluated by Nissl staining, some neurons appeared atrophic. We conclude that the induction of Fos-LI in neurons of the cerebral cortex and spinal cord is linked respectively to early onset-short stimulation and persistent excitatory or disinhibition phenomenon as a result of focal ischaemic brain injury. PMID- 9224839 TI - Planning an action. AB - The motor control of a sequence of two motor acts forming an action was studied in the present experiment. The two analysed motor acts were reaching-grasping an object (first target) and placing it on a second target of the same shape and size (experiment 1). The aim was to determine whether extrinsic properties of the second target (i.e. target distance) could selectively influence the kinematics of reaching and grasping. Distance, position and size of both targets were randomly varied across the experimental session. The kinematics of the initial phase of the first motor act, that is, velocity of reaching and hand shaping of grasping, were influenced by distance of the second target. No kinematic difference was found between movements executed with and without visual control of both hand and targets. These results could be due to computation of the general program of an action that takes into account extrinsic properties of the final target. Conversely, they could depend on a visual interference effect produced by the near second target on the control of the first motor act. In order to dissociate the effects due to second target distance from those due to visual interference, two control experiments were carried out. In the first control experiment (experiment 2) subjects executed movements directed towards spatial locations at different distances from the first target, as in experiment 1. However, the near second target was not presented and subjects were required to place the object on an arbitrary near position. Distance of the second (either real or arbitrary) target affected the reaching component of the first motor act, as in experiment 1, but not the grasp component. In the second control experiment (experiment 3), the pure visual interference effect was tested. Subjects were required to reach and grasp the object and to lift it in either presence or absence of a second near stimulus. No effect on the initial phase of the first motor act was observed. The results of the this study suggest a dissociation in the control of reaching and grasping, concerning not only visual analysis of extrinsic properties of the immediate target but also visual analysis of the final target of the action. In other words, the notion of modularity for the motor control can be extended to the construction of an entire action. PMID- 9224841 TI - The influence of movement segment difficulty on movements with two-stroke sequence. AB - Arm movements in the horizontal plane consisting of two segments were examined to determine whether the difficulty of the second segment influenced the kinematic characteristics of the first segment. The direction of the first segment was an elbow extension movement away from the trunk and remained constant throughout the experiment. The direction of the second segment varied between forearm extension and flexion movements. Based on Fitts' law, two different indexes of difficulty (ID) of the second segment were utilized by changing target size and movement amplitude. The effects of changing ID were examined for two different movement amplitudes. All movements were single-joint movements employing elbow flexion/extension and were recorded by an x-y digitizer. Variations in the ID of the second segment produced context-dependent kinematic changes in the performance of the initial segment. Movement duration increased when the ID was increased by reducing target size for both extension-extension sequence and extension-flexion sequences. Peak velocity also decreased for higher ID targets in the extension-flexion sequence. However, there was an interaction between the ID and movement amplitude in the extension-flexion sequence. In this sequence the duration of movement for the high ID/large movement amplitude condition increased substantially compared with the low ID/small movement amplitude condition. In addition, changing ID of the second segment influenced the time between the two segments (intersegment interval) in the extension-flexion sequence. Collectively, these data suggest that the planning of complex movements is based in part on the accuracy demands of multiple segments of the sequence. PMID- 9224842 TI - Movement-induced gain modulation of somatosensory potentials and soleus H reflexes evoked from the leg. I. Kinaesthetic task demands. AB - Movement-related gating of cerebral somatosensory evoked potentials (SEPs) occurs during active and passive movements of both the upper and the lower limbs. The general hypothesis was tested that the brain participates in setting the gain of the ascending path from somatosensory receptors of the human leg to the somatosensory cortex. In experiment 1, SEPs from Cz' and soleus H-reflexes were evoked by electrical stimulation of the tibial nerve in the popliteal fossa during passive movement about the right ankle. Early SEPs and H-reflexes sampled during simple passive movement were significantly attenuated when compared with stationary controls (P<0.05). The additional requirement of tracking the passive ankle movement with the other foot led to a significant relative facilitation of mean SEP, but not H-reflex amplitude, compared with means from passive movement alone (P<0.05). In experiment 2, SEPs were evoked in the active (tracking) leg during a forewarned reaction-time task. Subjects were required to move in a preferred direction or to track the passive movement of their right foot with their left. Significant attenuation of early SEP components occurred 100 ms prior to EMG onset (P<0.05), with no apparent effect due to tracking. In the 3rd experiment, SEPs and H-reflexes were evoked in the passively moved leg (the target for active movement of the left leg) during the same forewarned reaction time task. During the warning period, SEPs were significantly attenuated compared with stationary controls for non-tracking movements, but not for movements involving tracking (P<0.05). It is concluded that centrifugal factors are important in modulating SEP gain required by the kinaesthetic demands of the task. PMID- 9224843 TI - Movement-induced gain modulation of somatosensory potentials and soleus H reflexes evoked from the leg. II. Correlation with rate of stretch of extensor muscles of the leg. AB - Attenuation of initial somatosensory evoked potential (SEP) gain becomes more pronounced with increased rates of movement. Manipulation of the range of movement also might alter the SEP gain. It could alter joint receptor discharge; it should alter the discharge of muscle stretch receptors. We hypothesized that: (1) SEP gain reduction correlates with both the range and the rate of movement, and (2) manipulation of range and rate of movement to achieve similar estimated rates of stretch of a leg extensor muscle group (the vasti) results in similar decreases in SEP gain. SEPs from Cz', referenced to Fpz' (2 cm caudal to Cz and Fpz, respectively, according to the International 10-20 System), along with soleus H-reflexes were elicited by electrical stimulation of the tibial nerve at the popliteal fossa. Stable magnitudes of small M-waves indicated stability of stimulation. A modified cycle ergometer with an adjustable pedal crank and electric motor was used to passively rotate the right leg over three ranges (producing estimated vasti stretch of 12, 24 and 48 mm) and four rates (0, 20, 40 and 80 rpm) of movement. Two experiments were conducted. Ranges and rates of pedalling movement were combined to produce two or three equivalent estimated rates of tissue stretch of the vasti muscles at each of 4, 16, 32 and 64 mm/s. Tibial nerve stimuli were delivered when the knee was moved through its most flexed position and the hip was nearing its most flexed position. Means of SEP, H reflex and M-wave magnitudes were tested for rate and range effects (ANOVA). A priori contrasts compared means produced by equivalent estimated rates of vasti stretch. Increasing the rate of movement significantly increased the attenuation of SEP and H-reflex gain (P<0.05). Increasing the range of movement also significantly increased these gain attenuations (P<0.05). Combining these to achieve equivalent rates of stretch, through different combinations of rate and range, resulted in equivalent depressions of SEP gain. H-reflex gains were similarly conditioned. These results suggest that muscle stretch receptors play a more important role than joint or cutaneous receptors in regulating SEP gain consequent to movement. We note that the present calculation only considers the knee extensors; however, the biomechanical model of stretch applies also to receptors in the hip extensors. This paper and the companion one show that primary factors in the kinaesthetic components of the movement regulate activity induced gain attenuation of SEPs. PMID- 9224844 TI - Weak short-latency spinal projections to the long flexor of the human thumb. AB - The human thumb is controlled by a muscle, flexor pollicis longus (FPL), that is unique among mammals and contributes to manual dexterity. The present study sought to define whether the spinal reflex circuitry for this muscle differed from that for an adjacent muscle (flexor carpi radialis, FCR). In peri-stimulus time histograms, short-latency, largely monosynaptic excitation produced by median nerve stimulation was significantly less frequent and significantly smaller for FPL motor units than FCR motor units. Thus the motoneurone pools of adjacent muscles differ in their spinal reflex accessibility. The reflex control of FPL may thus be achieved by supraspinal pathways rather than the traditional monosynaptic arc. PMID- 9224845 TI - Tendon tap induces a single long-lasting excitatory reflex in the motoneurons of human soleus muscle. AB - The reflex responses of the soleus motor units to Achilles tendon taps were investigated. Two different techniques were used to analyse the motor unit data. The first approach was the classical technique which involved building peristimulus time histograms (PSTH) from the firing times of single motor units. The second approach was a relatively unused technique that involved plotting the instantaneous discharge frequency of the single motor unit against time (peristimulus frequencygram or PSF). Using PSTH as the indicator, we found that the tap to the tendon induced three separate reflex responses: the first response was a very short-lasting excitatory response or the jerk reflex, the second was a period of relative silence (silent period or the "trough"), and the third was a broad peak 170 ms after the stimulus. Using the same motor unit data, the PSF technique indicated that the tap to the tendon induced a single long-lasting excitatory reflex. The PSF displayed an increase starting from the latency of the jerk reflex and continuing for about 65 ms. There was no significant change in the discharge frequency at the end of the first excitatory response. Since the discharge frequency of a motoneuron has a strong positive linear relationship with the effective synaptic current it receives, it is suggested that throughout the 65-ms period the net (effective) synaptic drive to the soleus motoneurons was excitatory. It is therefore concluded that tendon tap induces a single long lasting excitatory reflex in the motoneurons of the soleus muscle. PMID- 9224847 TI - Viewing the hand prior to movement improves accuracy of pointing performed toward the unseen contralateral hand. AB - It is now well established that the accuracy of pointing movements to visual targets is worse in the full open loop condition (FOL; the hand is never visible) than in the static closed loop condition (SCL; the hand is only visible in static position prior to movement onset). In order to account for this result, it is generally admitted that viewing the hand in static position (SCL) improves the movement planning process by allowing a better encoding of the initial state of the motor apparatus. Interestingly, this wide-spread interpretation has recently been challenged by several studies suggesting that the effect of viewing the upper limb at rest might be explained in terms of the simultaneous vision of the hand and target. This result is supported by recent studies showing that goal directed movements involve different types of planning (egocentric versus allocentric) depending on whether the hand and target are seen simultaneously or not before movement onset. The main aim of the present study was to test whether or not the accuracy improvement observed when the hand is visible before movement onset is related, at least partially, to a better encoding of the initial state of the upper limb. To address this question, we studied experimental conditions in which subjects were instructed to point with their right index finger toward their unseen left index finger. In that situation (proprioceptive pointing), the hand and target are never visible simultaneously and an improvement of movement accuracy in SCL, with respect to FOL, may only be explained by a better encoding of the initial state of the moving limb when vision is present. The results of this experiment showed that both the systematic and the variable errors were significantly lower in the SCL than in the FOL condition. This suggests: (1) that the effect of viewing the static hand prior to motion does not only depend on the simultaneous vision of the goal and the effector during movement planning; (2) that knowledge of the initial upper limb configuration or position is necessary to accurately plan goal-directed movements; (3) that static proprioceptive receptors are partially ineffective in providing an accurate estimate of the limb posture, and/or hand location relative to the body; and (4) that static visual information significantly improves the representation provided by the static proprioceptive channel. PMID- 9224846 TI - In vitro evidence that the reduction in mesencephalic dopaminergic neurons in the weaver heterozygote is not due to a failure in target cell interaction. AB - The murine weaver (wv) mutation is characterized by a genetically determined loss of several neuronal populations, which include the nigrostriatal dopaminergic neurons. Animals homozygous for the wv gene exhibit marked deficits in dopaminergic morphological and neurochemical parameters. The wv gene shows incomplete dominance in that heterozygous (wv/+) mice exhibit moderate reductions in midbrain dopaminergic neuron number. It is unclear whether the dopaminergic neuronal loss in homozygous and heterozygous animals results from an effect of the wv gene solely on the dopaminergic neurons or is due to a failure of interaction of dopaminergic neurons with target cells of the striatum. This issue has been addressed utilizing three-dimensional reaggregate tissue cultures to determine whether the wv gene acts directly on the mesencephalic dopaminergic neurons. Embryonic mesencephalon and striatum from wv/+ and wild-type (+/+) brains were dissociated and the cells recombined into four mesencephalic-striatal aggregate combinations: (1) mesencephalic(+/+)-striatal(+/+)aggregates; (2) mesencephalic(wv/+)-striatal(wv/+)aggregates; (3) mesencephalic(wv/+) striatal(+/+)aggregates; and (4) mesencephalic(+/+)-striatal(wv/+)aggregates. At 29 days and 57 days of culture, the number of dopaminergic neurons and dopamine content from mesencephalic-striatal aggregates consisting of mixed genotype or from only wv/+ tissue were quantitated and compared with that from mesencephalic striatal cultures prepared from +/+ tissue alone. At both culture time points, aggregates containing wv/+ mesencephalon coaggregated with either wv/+ or +/+ striatum contained fewer dopaminergic neurons than mesencephalic-striatal cultures composed of only +/+ cells. Coaggregation of +/+ mesencephalon with wv/+ striatum did not have a detrimental effect on dopaminergic cell number. The findings demonstrate that the difference in the number of mesencephalic dopaminergic neurons between wv/+ and +/+ animals seen in vivo can be reproduced in three-dimensional reaggregate culture. Since the coculture of +/+ striatum with wv/+ mesencephalon did not appear to rescue wv/+ dopaminergic neurons in the aggregates as compared to wv/+ striatum and, wv/+ striatum proved to be a perfectly adequate target for +/+ mesencephalic dopaminergic neurons, it appears that the effect of the wv gene is on the dopaminergic neurons themselves. PMID- 9224848 TI - Sex differences in the response to GABA antagonists depend on the route of drug administration. AB - Sex differences in the responses to two GABA-related convulsants (bicuculline, picrotoxin) were studied in rats and mice following intraperitoneal (i.p.) and intravenous (i.v.) drug administration. Following i.p. administration male and female rats were equally sensitive to bicuculline, while female rats were more sensitive to picrotoxin. After i.v. infusion the threshold doses of bicuculline and picrotoxin producing running/bouncing clonus (RB clonus) were significantly lower in male than in female rats, i.e. male rats were more sensitive to both convulsants than females. Following i.p. administration, at some doses female mice were more sensitive to bicuculline and male mice to picrotoxin, although ED50 values between the sexes were not significantly different. After i.v. infusion, doses of bicuculline producing RB clonus and death were significantly lower in male than in female mice, i.e. male mice were more sensitive to bicuculline. The two sexes of mice were equally sensitive to i.v. administration of picrotoxin. While sex and species differences obtained following i.p. drug administration could presumably be explained by differences in pharmacokinetics, the i.v. route of drug administration is suggested as a reliable technique in the studies of sex and species differences in pharmacodynamics. PMID- 9224849 TI - Arm-movement-related neurons in the primate superior colliculus and underlying reticular formation: comparison of neuronal activity with EMGs of muscles of the shoulder, arm and trunk during reaching. AB - Neuronal activity was recorded from the superior colliculus (SC) and the underlying reticular formation in two monkeys during an arm reaching task. Of 744 neurons recorded, 389 (52%) clearly modulated their activity with arm movements. The temporal activity patterns of arm-movement-related neurons often had a time course similar to rectified electromyograms (EMGs) of particular muscles recorded from the shoulder, arm or trunk. These reach cells, as well as the muscles investigated, commonly exhibited mono- or biphasic (less frequently tri- or polyphasic) excitatory bursts of activity, which were related to the (pre )movement period, the contact phase and/or the return movement. The vast majority of reach cells exhibited a consistent activity pattern from trial to trial as did most of the muscles of the shoulder, arm and trunk. Similarities between the activity patterns of the neurons and the muscles were sometimes very strong and were especially notable with the muscles of the shoulder girdle (e.g. trapezius descendens, supraspinatus, infraspinatus or the anterior and medial deltoids). This high degree of co-activation suggests a functional linkage, though not direct, between the collicular reach cells and these muscles. Neuronal activity onset was compared with that of 25 muscles of the arms, shoulders and trunk. The majority of cells (78.5%) started before movement onset with a mean lead time of 149+/-90 ms, and 36.5% were active even before the earliest EMG onset. The neurons exhibited the same high degree of correlation (r=0.97, Spearman rank) between activity onset and the beginning of the arm movement as did the muscles (r=0.98) involved in the task. The mean neuronal reach activity (background subtracted) ranged between 7 and 193 impulses/s (mean 40.5+/-24.2). The mean modulation index calculated [(reach activity background activity)/reach activity+background activity)] was 0.75+/-0.23 for neurons (n=358) and 0.87+/ 0.14 for muscles (n=25). As the monkeys fixated the reach target constantly during an arm movement, neuronal activity which was modulated in this period was not related to eye movements. The three neck muscles investigated in the reach task exhibited no reach-related activity modulation comparable to that of either the reach cells or the muscles of the shoulder, arm and trunk. However, tonic neck muscle EMG was monotonically related to horizontal eye position. The clear skeletomotor discharge characteristics of arm-movement-related SC neurons revealed in this study agree with those already known from other sensorimotor regions (for example the primary motor, the premotor and parietal cortex, the basal ganglia or the cerebellum) and are consistent with the possible role of this population of reach cells in the control of arm movements. PMID- 9224850 TI - Anatomical distribution of arm-movement-related neurons in the primate superior colliculus and underlying reticular formation in comparison with visual and saccadic cells. AB - We recorded from 389 "reach" neurons (two monkeys) in the superior colliculus (SC) and underlying reticular formation (RF) or adjacent periaqueductal grey, whose activity was related to visually guided arm movements. Reach neurons were present from approximately 0.7 mm down to a depth of 6 mm below the surface of the SC (mean 3.7+/-1.3, n=389). Although this mean distribution was different from that of cells with visual (mean depth 1.7+/-1.4 mm, n=283) or saccadic responses (mean depth 2.0+/-1.4 mm, n=232), there was a large amount of overlap. Fifty-five per cent of all reach cells (213/389) were assumed to be located inside the SC. The others were considered to be located in the underlying RF. The characteristics of visual responses and saccadic bursts (e.g. response latencies, discharge rates, burst durations) of arm-movement-related neurons were not different from those of typical visual or saccade cells in the SC. Although reach neurons could be recorded in a large area of the SC, they were found more often in the lateral than in the medial parts (chi-squared=19.3, P<0.001). Possible pathways by which arm-movement-related neuronal activity in and below the SC might gain access to spinal motor structures are discussed. The location of arm movement-related neurons described in this study is in accordance with the known target areas of skeletomotor-related corticotectal projections and with the sites of origin of tectofugal pathways. It is concluded that this population of reach cells is in a position to relay and transmit limb movement information to the spinal motor system, where it may influence (or interact with) motor commands coming from other motor centres. PMID- 9224851 TI - Discrete and continuous planning of hand movements and isometric force trajectories. AB - We have previously demonstrated that, in preparing themselves to aim voluntary impulses of isometric elbow force to unpredictable targets, subjects selected default values for amplitude and direction according the range of targets that they expected. Once a specific target appeared, subjects specified amplitude and direction through parallel processes. Amplitude was specified continuously from an average or central default; direction was specified stochastically from one of the target directions. Using the same timed response paradigm, we now report three experiments to examine how the time available for processing target information influences trajectory characteristics in two-degree-of-freedom forces and multijoint movements. We first sought to determine whether the specification of force direction could also take the form of a discrete stochastic process in pulses of wrist muscle force, where direction can vary continuously. With four equiprobable targets (two force amplitudes in each of two directions separated by 22 degrees or 90 degrees), amplitude was specified from a central default value for both narrow and wide target separations as a continuous variable. Direction, however, remained specified as a discrete variable for wide target separations. For narrow target separations, the directional distribution of default responses suggested the presence of both discrete and central values. We next examined point-to-point movements in a multijoint planar hand movement task with targets at two distances and two directions but at five directional separations (from 30 degrees to 150 degrees separation). We found that extent was again specified continuously from a central default. Direction was specified discretely from alternative default directions when target separation was wide and continuously from a central default when separation was narrow. The specification of both extent and direction evolved over a 200-ms time period beginning about 100 ms after target presentation. As in elbow force pulses, extent was specified progressively in both correct and wrong direction responses through a progressive improvement in the scaling of acceleration and velocity peaks to the target. On the other hand, movement time and hand path straightness did not change significantly in the course of specification. Thus, the specification of movement time and linearity, global features of the trajectories, are given priority over the specific values of extent and direction. In a third experiment, we varied the distances between unidirectional target pairs and found that movement extent is specified discretely, like direction, when the disparity in distances is large. The implications of these findings for contextual effects on trajectory planning are discussed. The independence of extent and direction specification and the prior setting of response duration and straightness provide critical support for the hypothesis that point-to-point movements are planned vectorially. PMID- 9224852 TI - Corticospinal input in human gait: modulation of magnetically evoked motor responses. AB - Transcranial magnetic stimulation (TMS) of the motor cortex was applied during locomotion to investigate the significance of corticospinal input upon the gait pattern. Evoked motor responses (EMR) were studied in the electromyogram (EMG) of tibialis anterior (TA), gastrocnemius (GM) and, for reference, abductor digiti minimi (AD) muscles by applying below-threshold magnetic stimuli during treadmill walking in healthy adults. Averages of 15 stimuli introduced randomly at each of 16 phases of the stride cycle were analysed. Phase-dependent amplitude modulation of EMR was present in TA and GM which did not always parallel the gait-associated modulation of the EMG activity. No variation of onset latency of the EMR was observed. The net modulatory response was calculated by comparing EMR amplitudes during gait with EMR amplitudes obtained (at corresponding background EMG activities) during tonic voluntary muscle contraction. Large net responses in both muscles occurred prior to or during phasic changes of EMG activity in the locomotor pattern. This facilitation of EMR was significantly higher in leg flexor than extensor muscles, with maxima in TA prior to and during late swing phase. A comparison of this facilitation of TA EMR prior to swing phase and prior to a phasic voluntary foot dorsiflexion revealed a similar onset but an increased amount of early facilitation in the gait condition. The modulated facilitation of EMR during locomotion could in part be explained by spinal effects which are different under dynamic and static motor conditions. However, we suggest that changes in corticospinal excitability during gait are also reflected in this facilitation. This suggestion is based on: (1) the similar onset yet dissimilar size of facilitatory effects in TA EMR prior to the swing phase of the stride cycle and during a voluntary dynamic activation, (2) the inverse variation of EMR and EMG amplitudes during this phase, and (3) the occurrence of this inversion at stimulation strengths below motor threshold (motor threshold was determined during weak tonic contraction and EMR were facilitated during gait). It is hypothesized that the facilitation is phase linked to ensure postural stability and is most effective during the phases prior to and during rhythmical activation of the leg muscles resulting in anticipatory adjustment of the locomotor pattern. PMID- 9224853 TI - Cytotoxic lesions of the retrohippocampal region attenuate latent inhibition but spare the partial reinforcement extinction effect. AB - Experiment I assessed the effect of cytotoxic retrohippocampal (entorhinal and extra-subicular cortices) lesions on the development of latent inhibition (LI) using an off-the-baseline, between-subjects, conditioned emotional response paradigm. Sham-operated controls and unoperated rats that had been pre-exposed to a light stimulus prior to light-shock pairings showed less conditioned suppression towards the light stimulus than the nonpre-exposed animals, thus demonstrating LI. However, LI was not evident in rats with retrohippocampal lesions. In experiment 2, the same animals were trained to run in an straight runway for food. Half of the animals were trained under a 50% partial reinforcement schedule (i.e. they were rewarded randomly on half of the acquisition trials) and the other half were trained under a continuous reinforcement schedule (i.e. they were rewarded on every acquisition trial). When tested in extinction, animals trained on the partial reinforcement schedule showed greater persistence than animals trained on continuous reinforcement, thus demonstrating the partial reinforcement extinction effect (PREE). Rats with retrohippocampal lesions showed a PREE that was at least as clear as that seen in the sham-operated controls and in the unoperated animals. It is concluded that cytotoxic lesions of the retrohippocampal region selectively led to an abolition of LI, but spared the PREE. The present study thus provided evidence against the hypothesis that LI and the PREE share a common neural substrate. PMID- 9224854 TI - Kinematics of the freely moving head and neck in the alert cat. AB - In this study we examined connections between the moment-generating capacity of the neck muscles and their patterns of activation during voluntary head-tracking movements. Three cats lying prone were trained to produce sinusoidal (0.25 Hz) tracking movements of the head in the sagittal plane, and 22.5 degrees and 45 degrees away from the sagittal plane. Radio-opaque markers were placed in the cervical vertebrae, and intramuscular patch electrodes were implanted in five neck muscles, including biventer cervicis, complexus, splenius capitis, occipitoscapularis, and rectus capitis posterior major. Videofluoroscopic images of cervical vertebral motion and muscle electromyographic responses were simultaneously recorded. A three-dimensional biomechanical model was developed to estimate how muscle moment arms and force-generating capacities change during the head-tracking movement. Experimental results demonstrated that the head and vertebrae moved synchronously, but neither the muscle activation patterns nor vertebral movements were constant across trials. Analysis of the biomechanical model revealed that, in some cases, modification of muscle activation patterns was consistent with changes in muscle moment arms or force-generating potential. In other cases, however, changes in muscle activation patterns were observed without changes in muscle moment arms or force-generating potential. This suggests that the moment-generating potential of muscles is just one of the variables that influences which muscles the central nervous system will select to participate in a movement. PMID- 9224855 TI - Reference frames in saccadic targeting. AB - We attempt to determine the egocentric reference frame used in directing saccades to remembered targets when landmark-based (exocentric) cues are not available. Specifically, we tested whether memory-guided saccades rely on a retina-centered frame, which must account for eye movements that intervene during the memory period (thereby accumulating error) or on a head-centered representation that requires knowledge of the position of the eyes in the head. We also examined the role of an exocentric reference frame in saccadic targeting since it would not need to account for intervening movements. We measured the precision of eye movements made by human observers to target locations held in memory for a few seconds. A variable number of saccades intervened between the visual presentation of a target and a later eye movement to its remembered location. A visual landmark that allowed for exocentric encoding of the memory target appeared in half the trials. Variable error increased slightly with a greater number of intervening saccades. The landmark aided targeting precision, but did not eliminate the increase in variable error with additional intervening saccades. We interpret these results as evidence for a representation that relies on knowledge of eye position with respect to the head and not one that relies solely on updating in a retina-centered frame. Our results allow us to set an upper bound on the standard deviation of an eye position signal available to the saccadic system during short memory periods at 1.4 degrees for saccades of about 10 degrees. PMID- 9224856 TI - Common organization for unimanual and bimanual reach-to-grasp tasks. AB - In two experiments comparisons between characteristics of performance of a unimanual and a bimanual reach-to-grasp (prehension) task were made on an individual subject basis. The unimanual prehension task used required that the object be grasped by finger and thumb pad opposition, the bimanual task required that the grasp be made by opposing the pads on the two index fingers. Experiment 1 examined adaptation of prehension movements to objects of different size (width) but equal grasp surface area placed at different distances. Experiment 2 examined adaptation of movements to objects of different grasp surface areas. It was found that the aperture and transport components of the two prehension tasks developed over time in very similar fashion in all subjects. Movements were adapted to different task constraints in the same way as has previously been reported in the literature and were very similar in both tasks: maximum aperture increases with increasing object size and occurs later in the movement for larger objects; movement time increases with target distance; time of maximum aperture occurs earlier in the movement for targets with smaller grasp surface areas; movement times are longer for such objects, largely due to increases in the deceleration phase of the movement. These results support the notion that there is an effector independent level of organization that governs the coordination of movements during performance of reaching and grasping tasks. PMID- 9224857 TI - Sensory strategies in human postural control before and after unilateral vestibular neurotomy. AB - Vestibular inputs tonically activate the anti-gravitative leg muscles during normal standing in humans, and visual information and proprioceptive inputs from the legs are very sensitive sensory loops for body sway control. This study investigated the postural control in a homogeneous population of 50 unilateral vestibular-deficient patients (Meniere's disease patients). It analyzed the postural deficits of the patients before and after surgical treatment (unilateral vestibular neurotomy) of their diseases and it focused on the visual contribution to the fine regulation of body sway. Static posturographic recordings on a stable force-plate were done with patients with eyes open (EO) and eyes closed (EC). Body sway and visual stabilization of posture were evaluated by computing sway area with and without vision and by calculating the percentage difference of sway between EC and EO conditions. Meniere's patients were examined when asymptomatic, 1 day before unilateral vestibular neurotomy, and during the time-course of recovery (1 week, 2 weeks, 1 month, 3 months, and 1 year). Data from the patients were compared with those recorded in 26 healthy, age- and sex-matched participants. Patients before neurotomy exhibited significantly greater sway area than controls with both EO (+52%) and EC (+93%). Healthy participants and Meniere's patients, however, displayed two different behaviors with EC. In both populations, 54% of the subjects significantly increased their body sway upon eye closure, whereas 46% exhibited no change or significantly swayed less without vision. This was statistically confirmed by the cluster analysis, which clearly split the controls and the patients into two well-identified subgroups, relying heavily on vision (visual strategy, V) or not (non-visual strategy, NV). The percentage difference of sway averaged +36.7%+/-10.9% and -6.2%+/-16.5% for the V and NV controls, respectively; +45.9%+/-16.8% and -4.2%+/-14.9% for the V and NV patients, respectively. These two distinct V and NV strategies seemed consistent over time in individual subjects. Body sway area was strongly increased in all patients with EO early after neurotomy (1 and 2 weeks) and regained preoperative values later on. In contrast, sway area as well as the percentage difference of sway were differently modified in the two subgroups of patients with EC during the early stage of recovery. The NV patients swayed more, whereas the V patients swayed less without vision. This surprising finding, indicating that patients switched strategies with respect to their preoperative behavior, was consistently observed in 45 out of the 50 Meniere's patients during the whole postoperative period, up to 1 year. We concluded that there is a differential weighting of visual inputs for the fine regulation of posture in both healthy participants and Meniere's patients before surgical treatment. This differential weighting was correlated neither with age or sex factors, nor with the clinical variables at our disposal in the patients. It can be accounted for by a different selection of sensory orientation references depending on the personal experience of the subjects, leading to a more or less heavy dependence on vision. The change of sensory strategy in the patients who had undergone neurotomy might reflect a reweighting of the visual and somatosensory cues controlling balance. Switching strategy by means of a new sensory selection of orientation references may be a fast adaptive response to the lesion-induced postural instability. PMID- 9224858 TI - Glutamate receptor antagonists reduce spontaneous epileptiform activity in cortical wedges prepared from DBA/2 mice. AB - This study investigated the involvement of glutamatergic neurotransmission in the epileptiform activity demonstrated in cortical weges prepared from genetically audiogenic seizure-prone DBA/2 mice. Omission of Mg2+ from the perfusing medium initiated spontaneous epileptiform events with accompanying afterpotentials on the repolarizing phase. These spontaneous depolarizations also occurred in some 30% of the slices in the presence of Mg2+ (2 mM). The N-methyl-D-aspartate (NMDA) receptor antagonist 3-(2-carboxypiperazin-4-yl)-propenyl-1-phosphonic acid (CPP) and the non-competitive NMDA receptor channel blocker ketamine produced a significant reduction of these spontaneous depolarizations. 7-Chlorokynurenic acid (7-CKA), an antagonist at the strychnine-insensitive site on the NMDA receptor, also exerted an inhibitory effect. In addition the AMPA/kainate receptor antagonist 6,7-dinitroquinoxaline-2,3-dione (DNQX) suppressed the spontaneous events. These observations provide evidence that glutamatergic neurotransmission contributes to the epileptiform activity in this cortical preparation. PMID- 9224859 TI - Self-induced splitting of spiral-shaped spreading depression waves in chicken retina. AB - Spreading depression (SD) of electroencephalographic activity is a dynamic wave phenomenon in the central nervous system (CNS). The retina, especially the isolated chicken retina, is an excellent constituent of the CNS in which to observe the dynamic behavior of the SD wave fronts, because it changes its optical properties during a SD attack. The waves become visible as milky fronts on a black background. It is still controversial what the basic mechanistic steps of SD are, but certainly SD belongs to the self-organization phenomena occurring in neuronal tissue. In this work, spiral-shaped wave fronts are analyzed using digital video imaging techniques. We report how the inner end of the wave front, the spiral tip, breaks away repeatedly. This separation process is associated with a Z-shaped trajectory (extension approximately 1.2 mm) that is described by the tip over one spiral revolution (period 2.45+/-0.1 min). The Z-shaped trajectory does not remain fixed, but performs a complex motion across the retina with each period. This is the first time, to our knowledge, that established imaging methods have been applied to the study of the two-dimensional features of SD wave propagation and to obtaining quantitative data of their dynamics. Since these methods do not interfere with the tissue, it is possible to observe the intrinsic properties of the phenomenon without any external influence. PMID- 9224860 TI - Daily durations of spontaneous activity in cat's ankle muscles. AB - For an understanding of how various degrees of altered use (training, disuse) affect the properties of skeletal muscles, it is important to know how much they are used normally. The main aim of the present project was to produce such background knowledge for hindlimb muscles of the cat. In four adult female cats, each one being studied in several experimental sessions, ankle muscles were chronically implanted with electrodes for electromyographic (EMG) recording. The muscles recorded from were: extensor digitorum longus (EDL), peroneus longus (PL), tibialis anterior (TA), lateral gastrocnemius (LG) and soleus (SOL). For PL, TA and LG, there were anterior as well as posterior recording sites. During 24-h experimental sessions, the studied animal stayed, together with another cat, in a box large enough for playing and walking around. Using telemetric techniques, samples of EMG signals were recorded on tape for 4 min every 30 min. In an off-line analysis, measurements were made of the total accumulated duration of activity from each one of the studied muscle regions. These "duty times" were expressed as a percentage of total sampling duration. When averaged over the whole 24-h experimental period, the mean duty times per muscle region varied from 1.9% for EDL up to about 13.9% for SOL. Also, among predominantly fast muscles of mixed-fibre composition (i.e. all studied muscles except SOL), marked and statistically significant differences in duty time were found, mean values varying fivefold from 1.9% (EDL) to 9.5% (PL, posterior site). For all three muscles with simultaneous recordings from different sites, consistent and statistically significant differences in daily duty time were found between anterior and posterior regions (anterior less than posterior for TA and PL; anterior more than posterior for LG). We also measured the extent to which each 4 min sampling period was filled with activity (if any). As compared to muscles with a low mean 24-h duty time, those with high duty times were not active during more sampling periods per day, but, whenever being used, their activity lasted relatively longer. Such results were consistent with the view that differences in mean 24-h duty time might largely reflect differences in the extent to which the various muscles and muscle regions were used for long-lasting stabilizing contractions. PMID- 9224861 TI - The effect of fixation condition manipulations on antisaccade performance in schizophrenia: studies of diagnostic specificity. AB - This series of studies evaluated (1) hypotheses that poor antisaccade performance is attributable to confounding variables (e.g., visual attention deficits, incomplete understanding of task demands) and (2) the specificity of poor antisaccade performance to schizophrenia. In addition to self-correcting errors before being cued to do so, schizophrenia patients also showed the expected saccadic reaction time changes to fixation condition manipulations: decreased latencies for gap and increased latencies for overlap trials. These data suggest that schizophrenia patients are adequately engaged in and understand the antisaccade task. Schizophrenia patients made fewer correct antisaccade responses than other psychiatric patients (obsessive-compulsive and bipolar disorder) and normal subjects. The first-degree relatives of schizophrenia patients also generated a decreased proportion of correct antisaccade responses compared with normal subjects. For schizophrenia patients who performed below the range of normal subjects, 26% of their relatives also performed below the normal range. Conversely, patients who performed normally did not have a single poor-performing relative. These data suggest that increased antisaccade error rates may index a liability for schizophrenia within a subset of families. PMID- 9224862 TI - Control of frontal plane body motion in human stepping. AB - During a step the body's centre of mass (CoM) typically remains medial to the supporting foot and therefore the body is unstable and falling (sideways) under gravity. This may make it difficult to adjust the frontal-plane body motion appreciably once the step is under way. We have therefore investigated whether this motion could be controlled largely in a ballistic manner, that is by setting the initial (toe-off) position and velocity of the CoM such that the fall develops as required for the particular step without the need for appreciable mid step adjustment. Subjects stepped in different directions and from different postures, and the resulting motion of their CoM in the frontal plane was compared with that of a single-segment mathematical model of the body which falls freely under the influence of gravity. The lateral position and velocity of subjects' CoM at toe-off varied across the different step types in a manner consistent with a ballistic mode of control. Furthermore the model, given these positions and velocities as initial conditions, closely predicted the subsequent CoM motion. The results suggest that subjects may produce the different body trajectories required for different types of step largely in a ballistic manner. This would imply that the central nervous system must judge in advance the size and direction of the initial "throw" given to the body-mass. PMID- 9224863 TI - Role of primary somatic sensory cortex in the categorization of tactile stimuli: effects of lesions. AB - We lesioned the right primary somatic sensory (S1) cortex in two monkeys trained to categorize the speed of moving tactile stimuli. Animals performed the task by pressing with the right hand one of two target switches to indicate whether the speed of a probe moving across the glabrous skin of the left hand was low or high. Sensory performance was evaluated with psychometric techniques and motor behavior was monitored by measuring the reaction (RT) and movement (MT) times before the experiment and throughout the 60 days after the ablation of SI cortex. After the lesion, there was a slight increase in the RTs but no change in the MTs, indicating that removal of SI cortex did not affect the animals' capacity to detect the stimuli. However, monkeys lost their ability to categorize the stimulus speeds. This effect was observed from the 1st day after the lesion until the end of the study. We conclude that somatosensory areas outside SI can by themselves process tactile information in a limited way and that the extraction of higher-order features that takes place during the categorization task requires the intervention of SI cortex. PMID- 9224864 TI - Circular trajectory formation during blind locomotion: a test for path integration and motor memory. AB - Eight healthy subjects were asked to walk blindfolded along circular paths of different radii after several practice trials with vision. Their task was to stop after completing two full revolutions. They always walked counter-clockwise (CCW) in (a) a control condition (CONTROL), including the instructions mentioned above, (b) with the further instruction to count backwards in twos (MENTAL), (c) with the instruction to count loudly (LOUD). The movement of two markers lying along the head naso-occipital axis was recorded by means of an ELITE system. Total walked distance (DISTANCE), total head turning angle (ANGLE) and average radius (RADIUS) of the trajectories performed were measured. All subjects were able to perform approximately circular trajectories. They consistently overshot the ideal radius independently of the condition and circle size, undershot the total angle and overshot total distance. The LOUD condition induced greater errors in the performance but only on total distance (P<0.05). A strong correlation was found between the errors in radius and total distance but not between distance and total angle. Principal components analysis suggested that radius and distance share a common source of errors while total angle produced independent errors. The results indicate that (a) circular trajectories can be generated starting from spatial and/or motor memory, without the aid of visual information; (b) the task needs some attentional control and does not involve simple automatic processing of afferent information; (c) different sensory information or different processing modes are probably involved in the estimation of the curvature and length of the walked path on the one hand, and of the total rotation angle on the other. PMID- 9224865 TI - Distinct substance P- and calretinin-containing projections from the supramammillary area to the hippocampus in rats; a species difference between rats and monkeys. AB - Our recent studies showed the co-existence of substance P and calretinin in the supramammillo-hippocampal pathway of monkeys, as well as species differences in the synaptic targets of extrinsic substance P fibers in the hippocampi of monkeys and rats. Experiments used: (1) single and multiple stereotaxic injection of wheat germ agglutinin-conjugated HRP into the hippocampus and immunostaining for substance P in the supramammillary area; (2) colocalization of substance P and calretinin in supramammillary area cells; and (3) colocalization of these two neurochemicals in retrogradely labeled supramammillary projective cells of both male and female rats. These demonstrated: (a) many calretinin- and fewer substance P-immunoreactive neurons retrogradely labeled in the ipsilateral supramammillary area; (b) approximately 74% of all substance P cells contain calretinin and 9% of the calretinin neurons co-contain substance P; and, most importantly (c) none of the retrogradely labeled supramammillary cells colocalize calretinin and substance P. These results indicate the presence of two distinct supramammillo-hippocampal projections in the rat, one that contains substance P and the other calretinin. The latter innervates the same areas as those in the monkey, and the former terminates only in the CA2 hippocampal subfield. PMID- 9224866 TI - The effects of disorientation on visual landmark control of head direction cell orientation. AB - Head direction (HD) and place cells were recorded in rats that had previously exhibited significant acquisition deficits on a radial arm maze task following disorientation treatment. In this study we determined whether this behavioral impairment was associated with a lack of landmark stimulus control over the preferred orientations of HD and place cells. Neurons were recorded as animals retrieved food pellets in a cylindrical apparatus containing a single cue card. Some of these HD cells were also recorded while animals explored an eight-arm radial maze in a similar cue-controlled environment. The stimulus control of the landmarks in each environment was assessed by rotating the landmark and examining the subsequent preferred orientations of HD and place cells. Animals underwent disorientation treatment before and after each recording session. Despite this disorientation, rotation of the cue card in the cylindrical apparatus resulted in a corresponding shift in the preferred orientations of HD and place cells in 13 of 15 and 7 of 7 recording sessions, respectively. On the radial arm maze, rotation of the landmark cue was associated with a corresponding shift in the HD cell's preferred orientation in 7 of 9 sessions. These results suggest that a visual landmark's stimulus control may not require a learned association between that landmark and an animal's stable experience in an environment. Furthermore, instability in the HD cell system is unlikely to account for the impaired performance of the disoriented animals in the radial arm maze. Rather, these impairments may be due to the animal's inability to utilize stable representations of the environment provided by HD and place cells. PMID- 9224868 TI - Protein secretion by Gram-negative bacterial ABC exporters--a review. AB - One of the strategies used by Gram-negative bacteria to secrete proteins across the two membranes which delimit the cells is sec-independent and dedicated to proteins lacking an N-terminal signal peptide. Most of these proteins display a C terminal secretion signal located in the last 60 amino acids (aa). Using one Erwinia chrysanthemi protease, PrtG, secreted by such a pathway it was shown that the smallest C-terminal sequence allowing efficient secretion contains the last 29 aa of PrtG and that low but significant secretion can be promoted by the last 15 aa of PrtG. Moreover, the extreme C-terminal motif, consisting of a negatively charged aa followed by several hydrophobic aa must be exposed and is conserved amongst many proteins following this pathway. This secretion system depends on ABC protein-mediated exporters, which consist of three cell envelope proteins: two inner membrane proteins, an ATPase (the ABC protein), a membrane fusion protein (MFP) and an outer membrane polypeptide. These Gram-negative bacterial protein exporters are dedicated to the secretion of one or several closely related proteins belonging to the toxin, protease and lipase families. The genes encoding the three secretion proteins and the exoproteins are usually all linked, consistent with the specificity of the systems. Er. chrysanthemi metalloproteases B and C and Serratia marcescens hemoprotein HasA are secreted by such homologous pathways and interact with the ABC protein. Interaction between the ABC protein and its substrate has also been evidenced by studies on protease and HasA hybrid transporters obtained by combining components from each system. Association between hemoprotein HasA and the three exporter secretion proteins was demonstrated by affinity chromatography on hemin agarose on which the substrate remained bound with the three secretion proteins. The three components' association was ordered and substrate binding was required for the formation of this multiprotein complex. PMID- 9224869 TI - Recent progress and future directions in studies of the main terminal branch of the general secretory pathway in Gram-negative bacteria--a review. AB - The main terminal branch (MTB) of the general secretory pathway is used by a wide variety of Gram- bacteria to transport exoproteins from the periplasm to the outside milieu. Recent work has led to the identification of the function of two of its 14 (or more) components: an enzyme with type-IV prepilin peptidase activity and a chaperone-like protein required for the insertion of another of the MTB components into the outer membrane. Despite these important discoveries, little tangible progress has been made towards identifying MTB components that determine secretion specificity (presumably by binding to cognate exoproteins) or which form the putative channel through which exoproteins are transported across the outer membrane. However, the idea that the single integral outer membrane component of the MTB could line the wall of this channel, and the intriguing possibility that other components of the MTB form a rudimentary type-IV pilus like structure that might span the periplasm both deserve more careful examination. Although Escherichia coli K-12 does not normally secrete exoproteins, its chromosome contains an apparently complete set of genes coding for MTB components. At least two of these genes code for functional proteins, but the operon in which twelve of the genes are located does not appear to be expressed. We are currently searching for conditions which allow these genes to be expressed with the eventual aim of identifying the protein(s) that E. coli K 12 can secrete. PMID- 9224870 TI - Filamentous phage assembly: variation on a protein export theme. AB - Biogenesis of both filamentous phage and type-IV pili involves the assembly of many copies of a small, integral inner membrane protein (the phage major coat protein or pilin) into a helical, tubular array that passes through the outer membrane. The occurrence of related proteins required for assembly and export in both systems suggests that there may be similarities at the mechanistic level as well. This report summarizes the properties of filamentous phage and the proteins required for their assembly, with particular emphasis on features they may share with bacterial protein export and pilus biogenesis systems, and it presents evidence that supports the hypothesis that one of the phage proteins functions as an outer membrane export channel. PMID- 9224871 TI - Biogenesis of the bundle-forming pilus of enteropathogenic Escherichia coli: reconstitution of fimbriae in recombinant E. coli and role of DsbA in pilin stability--a review. AB - Enteropathogenic Escherichia coli (EPEC) adhere to tissue culture cells in a distinct pattern known as localized adherence (LA). We have defined two loci necessary for LA. A plasmid-encoded gene cluster encodes bundlin, the major structural subunit of a type-IV fimbria called the bundle-forming pilus (BFP), a prepilin peptidase necessary for processing of pre-bundlin to its mature form, and twelve other proteins. Under the control of an exogenous promoter, these 14 genes are sufficient for the biogenesis of BFP in a heterologous E. coli host. The chromosomal gene dsbA, which encodes a periplasmic disulfide-bond oxidoreductase, is also required for LA. In the absence of DsbA protein, bundlin is made but rapidly degraded. Pre-bundlin is also rapidly degraded in the absence of DsbA, suggesting that the prepilin is a transcytoplasmic protein simultaneously accessible to enzymes on both sides of the inner membrane. These studies offer a fresh perspective on the biogenesis of type-IV pili. PMID- 9224872 TI - Longus pilus of enterotoxigenic Escherichia coli and its relatedness to other type-4 pili--a minireview. AB - Longus is a long pilus produced by human enterotoxigenic Escherichia coli (ETEC) which shares significant structural and biochemical features with class-B type-4 pili. These pili include the toxin-coregulated pilus (TCP) of Vibrio cholerae, the bundle-forming pilus (BFP) of enteropathogenic E. coli and both longus and the colonization factor antigen III (CFA/III) of ETEC. These pili are produced under defined growth conditions indicating that they are under the control of different regulatory elements. While TCP is chromosomally encoded, the remaining pili are encoded on large virulence plasmids. Longus and CFA/III are closely related pili although certain DNA and protein differences also exist between them. This may account for the differences in the regulation, surface presentation, antigenicity, and prevalence of these two pilins among ETEC. Neighboring lngA, a second open reading frame termed lngB was found which encodes a protein with significant homology to proteins which are part of a type-II secretory system such as XcpV, OutC, and PulO of Pseudomonas aeruginosa, Erwinia chrysanthemi, and Klebsiella pneumoniae, respectively. This suggests that lngB may be an accessory gene involved in biogenesis of longus. PMID- 9224873 TI - The conserved tetracysteine motif in the general secretory pathway component PulE is required for efficient pullulanase secretion. AB - The PulE component of the pullulanase secretion pathway, a typical main terminal branch of the general secretory pathway, has a tetracysteine motif (4Cys) that is also present in almost all of the many PulE homologues, including those involved in type-IV piliation and conjugal DNA transfer. The 4Cys resembles a zinc-binding motif found in other proteins such as adenylate kinases, which may be pertinent in view of the fact that PulE has a consensus ATP-binding motif and since at least one PulE homologue has been reported to have kinase activity. In PulE, the Cys residues of this motif form scrambled intra- and intermolecular disulfide bonds when cells are disrupted. Replacement of one or more Cys of this motif by Ser reduces PulE function, but at least two adjacent Cys must be replaced to prevent intramolecular disulfide bond formation. PMID- 9224874 TI - The invasion-associated type-III protein secretion system in Salmonella--a review. AB - The genetic determinants that confer upon Salmonella the ability to enter non phagocytic cells are largely encoded in a pathogenicity island located at centisome 63 of the bacterial chromosome. Molecular genetic analysis has revealed that this region encodes a specialized protein secretion system that mediates the export and/or translocation of putative signaling proteins into the host cell. This protein secretion system, which has been termed type III or contact dependent, has also been identified in other plant and animal pathogens that have, in common, the ability to interact with eukaryotic host cells in an intimate manner. PMID- 9224875 TI - The tcp gene cluster of Vibrio cholerae. AB - The toxin co-regulated pilus (TCP) has been identified as a critical colonization factor in both animal models and humans for Vibrio cholerae O1. The major pilin subunit, TcpA (and also TcpB), is similar to type-4 pilins but TCP probably more appropriately belongs to a sub-class which includes the bundle-forming pilus of enteropathogenic Escherichia coli. The genes for TCP biosynthesis and assembly are clustered with the exception of housekeeping functions such as TcpG (=DsbA, a periplasmic disulfide bond epimerase). The nt sequences from El Tor and classical strains show only minor differences corresponding to the major regulatory regions and in TcpA itself. These differences are thought to account for the alternate conditions required for expression of TCP by the two biotypes and the antigenic variation and lack of cross-protection. Aside from the TcpA only a few of the proteins have had their roles in TCP biogenesis defined. Regulation of TCP is controlled by the ToxR regulon via ToxT with a possible involvement of TcpP and the cAMP-CRP system. Experiments using the infant mouse cholera model have now shown that TCP is a colonization factor and protective antigen for both classical and El Tor O1 strains and in the O139 Bengal serotype and that the mannose sensitive haemagglutinin pilus does not appear to play a comparable role. PMID- 9224876 TI - Translocation failure in a type-4 pilin operon: rfb and tcpT mutants in Vibrio cholerae. AB - Defined chromosomal mutations that lead to assembly failure of the toxin coregulated pilus (TCP) of Vibrio cholerae provide useful insights into the biogenesis of a type-4 pilus. Mutants in rfb affecting LPS O-antigen biosynthesis, and strains depleted of the cytoplasmic membrane-associated ATP binding protein TcpT, provide contrasting TCP export-defective phenotypes acting at different locations. Mutants in the perosamine biosynthesis pathway of V. cholerae 569B result in an rfb phenotype with an LPS consisting only of core oligosaccharide and lipid A. Such strains are unable to assemble TCP, and TcpA subunits are found in the periplasm and membrane fractions. In both rfb and tcpT mutants, the export defect is specific and complete. TcpT is a member of a large family of cytoplasmic membrane-associated ATP-binding proteins which are essential in type-4 pilin systems and in many non-pilin outer membrane transporters in Gram-negative bacteria. The behaviour of translocation-arrested TcpA in rfb and tcpT mutants is indistinguishable from that within assembled pilus under a range of conditions including flotation in density gradients, chemical cross-linking, and detergent extraction experiments. From the data presently available, it would appear that TcpA requires TcpT-mediated translocation from the cytoplasmic membrane and that TcpT stabilizes the subunit at or immediately beyond this stage, before crossing the outer membrane. PMID- 9224877 TI - Domains within the Vibrio cholerae toxin coregulated pilin subunit that mediate bacterial colonization. AB - Several experimental approaches have provided evidence suggesting that a domain within the C-terminal region of the TcpA pilin, delineated by the single disulfide loop, is directly responsible for the colonization function mediated by the toxin coregulated pilus (TCP) of Vibrio cholerae. This evidence includes the mapping of domains recognized by protective monoclonal antibodies to this region, the ability of peptides from within this region to elicit cholera protective antibody, the construction of tcpA missense mutations that abolish TCP function, and the requirement of a periplasmic disulfide isomerase to produce functional TCP. PMID- 9224878 TI - Genes involved in the biogenesis and function of type-4 fimbriae in Pseudomonas aeruginosa. AB - Type-4 fimbriae are filamentous polar organelles which are found in a wide variety of pathogenic bacteria. Their biogenesis and function is proving to be extremely complex, involving the expression and coordinate regulation of a large number of genes. Type-4 fimbriae mediate attachment to host epithelial tissues and a form of surface translocation called twitching motility. In Pseudomonas aeruginosa they also appear to function as receptors for fimbrial-dependent bacteriophages. Analysis of mutants defective in fimbrial function has allowed the identification of many of the genes involved in the biogenesis of these organelles. Thus far over 30 genes have been characterized, which fall into two broad categories: those encoding regulatory networks that control the production and function of these fimbriae (and other virulence determinants such as alginate) in response to alterations in environmental conditions; and those encoding proteins involved in export and assembly of these organelles, many of which are similar to proteins involved in protein secretion and DNA uptake. These systems all appear to be closely related and to function in the assembly of surface-associated protein complexes that have been adapted to different biological functions. PMID- 9224879 TI - The type-4 pilus is the major virulence-associated adhesin of Pseudomonas aeruginosa--a review. AB - Pseudomonas aeruginosa (Pa) produces several surface-associated adherence factors or adhesins which promote attachment to epithelial cells and contribute to the virulence of this pathogen. Among them, the type-4 pilus accounts for about 90% of the adherence capability of Pa to human lung pneumocyte A549 cells. Furthermore, it is responsible for more than 90% of the virulence in AB.Y/SnJ mice. Pa type-4 pili display a tip-base differentiation with the adherence function located at the tip of the pilus. All Pa pili prototypes characterized so far contain an intrachain disulfide loop (DSL) of 12 to 17 semi-conserved amino acid residues at the C-terminus of pilin. In Pa, this DSL comprises the epithelial cell-binding domain. Despite little sequence homology, DSL-containing peptides of different pilin prototypes seemingly reveal striking structural similarities. Two beta-turns within the loop and the disulfide bridge impose significant structural rigidity on the DSL pilin peptide, suggesting a conformationally conserved binding domain. Insertions of C-terminal pilin peptides with disrupted DSL displayed on the surface of bacterial S-layer mediate the same receptor binding characteristics as pili, indicating that a DSL is not essential in maintaining the functionality of the binding domain. Pa pili bind specifically to the carbohydrate moiety of the glycosphingolipids (GSL) asialo G(M1) and asialo-G(M2) and, to a much weaker extent, to lactosyl ceramide and ceramide trihexoside. The disaccharide sequence GalNAc beta(1-4)Gal, common in both asialo-G(M1) and asialo-G(M2), likely represents the minimal structural receptor motif recognized by the pili. Pa pili also bind to surface-localized proteins of human epithelial cells and other cell types, suggesting that non sialylated GSL and (glyco)proteins function as receptors of pili. In addition to the major pilus adhesin, exoenzyme S and, as recent studies indicate, flagella, are further protein adhesins of Pa with GSL receptor binding specificities similar to those of pili. PMID- 9224880 TI - Molecular genetic analysis of type-4 pilus biogenesis and twitching motility using Pseudomonas aeruginosa as a model system--a review. AB - Genetic analysis of Pseudomonas aeruginosa pilus biogenesis and twitching motility has revealed the requirement for several pil loci which have been localized to different regions of the chromosome. One pil locus, designated pilE, resides at approx. 71 min on the PAO genetic map, a region of the chromosome previously shown to harbor a number of genes required for pilus assembly (i.e., pilA, -B, -C, -D, -R and -S). The PilE protein shows significant sequence identity to the N-terminal domain of PilA as well as to the pilin precursors from a variety of type-4 pilus producers. Included within this homologous region is a short, positively charged leader sequence followed by a prepilin peptidase cleavage site and a largely hydrophobic region. Additionally, an unlinked set of pil genes, designated pilG, -H, -I, -J and -K, has been localized to the SpeI fragment H which corresponds to approx. 20 min on the PAO genetic map. This gene cluster encodes proteins that demonstrate remarkable similarity to the chemotaxis proteins of enterics and the gliding bacterium Myxococcus xanthus and are thought to be part of a signal transduction system that controls P. aeruginosa pilus biosynthesis and twitching motility. PMID- 9224881 TI - Structure-function relationship of type-IV prepilin peptidase of Pseudomonas aeruginosa--a review. AB - The bifunctional enzyme prepilin peptidase (PilD) from Pseudomonas aeruginosa is a key determinant in both type-IV pilus biogenesis and extracellular protein secretion, in its roles as a leader peptidase and MTase. It is responsible for endopeptidic cleavage of the unique leader peptides that characterize type-IV pilin precursors, as well as proteins with homologous leader sequences that are essential components of the general secretion pathway found in a variety of Gram negative pathogens. Following removal of the leader peptides, the same enzyme is responsible for the second posttranslational modification that characterizes the type-IV pilins and their homologues, namely N-methylation of the newly exposed N terminal amino acid residue. This review discusses some of the work begun in order to answer questions regarding the structure-function relationships of the active sites of this unique enzyme. PMID- 9224882 TI - Transformation competence and type-4 pilus biogenesis in Neisseria gonorrhoeae--a review. AB - In Neisseria gonorrhoea (Ngo), the processes of type-4 pilus biogenesis and DNA transformation are functionally linked and play a pivotal role in the life style of this strictly human pathogen. The assembly of pili from its main subunit pilin (PilE) is a prerequisite for gonococcal infection since it allows the first contact to epithelial cells in conjunction with the pilus tip-associated PilC protein. While the components of the pilus and its assembly machinery are either directly or indirectly involved in the transport of DNA across the outer membrane, other factors unrelated to pilus biogenesis appear to facilitate further DNA transfer across the murein layer (ComL, Tpc) and the inner membrane (ComA) before the transforming DNA is rescued in the recipient bacterial chromosome in a RecA-dependent manner. Interestingly, PilE is essential for the first step of transformation, i.e., DNA uptake, and is itself also subject to transformation-mediated phase and antigenic variation. This short-term adaptive mechanism allows Ngo to cope with changing micro-environments in the host as well as to escape the immune response during the course of infection. Given the fact that Ngo has no ecological niche other than man, horizontal genetic exchange is essential for a successful co-evolution with the host. Horizontal exchange gives rise to heterogeneous populations harboring clones which better withstand selective forces within the host. Such extended horizontal exchange is reflected by a high genome plasticity, the existence of mosaic genes and a low linkage disequilibrium of genetic loci within the neisserial population. This led to the concept that rather than regarding individual Neisseria species as independent traits, they comprise a collective of species interconnected via horizontal exchange and relying on a common gene pool. PMID- 9224883 TI - Microevolution and epidemic spread of serogroup A Neisseria meningitidis--a review. AB - An extensive and representative strain collection of serogroup A Neisseria meningitidis was established. These bacteria were obtained from different endemic and epidemic/pandemic sources and include strains from diseased patients and healthy carriers. The genetic relationships of the bacteria were defined by multi locus enzyme electrophoresis and sequence polymorphisms of genetically variable antigens have been analyzed in closely-related groupings. The results are interpreted as reflecting a balance of recombination events, which disrupt clonal relationships, and sequential bottlenecks, which purify the bacterial population of genetic variants during epidemic spread. PMID- 9224884 TI - Post-translational modifications of meningococcal pili. Identification of common substituents: glycans and alpha-glycerophosphate--a review. AB - Neisseria meningitidis elaborate filamentous adhesins, pili or fimbriae, which belong to the type-4 structural group of pili also found on other bacterial pathogens such as Neisseria gonorrhoeae, Pseudomonas aeruginosa, Moraxella bovis and Dichelobacter nodosus. Meningococcal pili readily undergo structural variations which arise as a result of inter- and intra-genomic recombinational events. Structural variations often result in variations in bacterial adhesion mediated by pili. Studies on structure/function relationship of meningococcal pili have shown that pili are post-translationally modified and contain unusual covalently-linked substituents. The presence of glycans was shown by the use of specific carbohydrate labelling, chemical deglycosylation and by introducing galE mutations; the latter studies confirmed the presence of galactose on pili. Mass spectrometric and other analysis of pilin-derived tryptic peptides and linked substituents were used to determine the structure of an unusual O-linked trisaccharide, Gal beta1-4 Gal alpha1-3[2,4-diacetamido-2,4,6-trideoxyhexose]. Similar studies have also identified, perhaps a unique substituent, alpha glycerophosphate, which is attached to Ser93 by a phosphodiester linkage. PMID- 9224885 TI - Type-4 pili and meningococcal adhesiveness. AB - The ability to interact with non-phagocytic cells is a crucial virulence attribute of the meningococcus. Pili play a major role in this process and are the only means yet discovered by which capsulated bacteria may adhere to cells. Pilus-mediated adhesion is a two-step process which requires (i) the expression of the adhesin PilC1 and (ii) the expression of an appropriate pilin variant. Some pilin variants have the ability to modify the degree of adhesiveness through the formation of bundles of pili which increases bacteria-bacteria interactions. PMID- 9224886 TI - The pilus colonization factor of pathogenic neisserial species: organelle biogenesis and structure/function relationships--a review. AB - Type-IV pilus expression plays a critical role in the interactions between Neisseria gonorrhoeae, Neisseria meningitidis and their human host. We have focused on experiments designed to elucidate the mechanisms of organelle biogenesis as one means of understanding the complexities of pilus biology in these species. Employing a variety of approaches, genes and gene products essential to pilus biogenesis have been identified and characterized. The findings indicate that the neisserial type-IV pilus biogenesis machinery is most closely related to that operating in Pseudomonas aeruginosa and other pseudomonad species. This interrelatedness is documented at the levels of gene organization, DNA homologies and identities between the primary structures of the components. Despite these similarities, the biological correlates of pilus expression in the pathogenic Neisseria are quite unique. The current status of our embryonic understanding of the factors influencing organelle biogenesis is presented. In the context of this workshop, emphasis has been placed on specific contributions made through studies of gonococci and meningococci to the field as a whole.. PMID- 9224887 TI - Type-4 pilus-structure: outside to inside and top to bottom--a minireview. AB - We have recently proposed a computational model of the N. gonorrhoeae pilus fiber based on the high resolution X-ray crystal structure of the component protein pilin, combined with available biophysical and genetic data [Parge et al. (1995) Nature 378, 32-38]. In parallel, we have used anti-peptide antibodies to distinguish buried and exposed regions of pilin within the assembled fiber [Forest et al. (1996) Infect. Immun. 64, 644-652]. This mini-review addresses the properties of the current pilus model and the locations of end-exposed epitopes. The fiber forms a three-layered structure of coiled conserved alpha helices surrounded by beta-sheet, with the hypervariable region as the most highly exposed portion. Overall the pilus model developed from diffraction and antibody mapping is expected to be representative of type-4 pili with general implications for type-4 assembly, function, and interactions with other proteins and cell membranes. PMID- 9224888 TI - Type-4 pili of Kingella denitrificans. AB - Kingella denitrificans possess type-4 pili, and the type strain, ATCC 33394, contains at least four complete copies of type-4 pilin-encoding genes. Previously reported hybridization patterns of K. denitrificans chromosomal DNA seen using a Neisseria gonorrhoeae pilin gene region probe, had been interpreted as representing possible partial, silent gene loci. This now appears to be due to cross-reaction to multiple copies of 18-bp inverted repeat structures. Data are presented on a variety of colony variants which have changed from a spreading corroding (SC) phenotype to a nonspreading-noncorroding (N) phenotype. Interestingly, while the SC to N transition is most often associated with loss of piliation in other bacteria containing type-4 pili, many of the K. denitrificans N variants still produce pilin, and some still produce pili. PMID- 9224889 TI - Uptake and processing of DNA by Acinetobacter calcoaceticus--a review. AB - In natural transformation, DNA in the form of macromolecular fragments can be translocated across the cell envelope of prokaryotic microorganisms. During the past two decades, several, largely mutually contradictory, hypotheses have been forwarded to explain the molecular mechanism and bioenergetics of this translocation process. Other biomacromolecules are translocated across the bacterial cell envelope as well, such as polysaccharides and proteins, the latter for instance in the process of the assembly of type-IV pili. This brings up the question whether or not common components are involved. Here, we review analyses of DNA translocation in Acinetobacter calcoaceticus, a Gram-negative eubacterium that is able to migrate through twitching motility, and also shows a high frequency of natural transformation. DNA uptake in this organism is an energy dependent process. Upon entry into the cells, the DNA fragments are integrated into the resident chromosome when a sufficiently large region of mutual homology is available (200 to 400 bp). However, this process is rather inefficient, and on the average 500 bp of each incoming fragment is degraded through exonuclease activity. Upon covalent attachment of a bulky protein molecule to the transforming DNA, the DNA-translocation machinery becomes blocked in further translocation activity. Since A. calcoaceticus is not well suited for transposon mutagenesis, a random mutagenesis procedure has been developed, based on the ligation of an antibiotic-resistance marker to random fragments of chromosomal DNA. This method was used to generate several mutants impaired in the natural transformation process. Three of these have been characterized in detail. No components, common to the translocation of macromolecules through the cell envelope of Acinetobacter, have been detected in this screen. PMID- 9224890 TI - Binding and transport of transforming DNA by Bacillus subtilis: the role of type IV pilin-like proteins--a review. AB - The pathway for binding, processing and transport of transforming DNA into competent cells of Bacillus subtilis is described. The known proteins involved in mediating these processes are reviewed in turn, including several that resemble proteins required for type-IV pilus assembly and function, and those involved in protein secretion. Based on the phenotypes of null mutations in the cognate genes for these proteins, on similarities to other proteins and on membrane localization and topology data, proposals are made for the roles of the individual proteins in the transformation process. A dynamic model is suggested for the presentation of transforming DNA to the transport apparatus. PMID- 9224891 TI - The DAN1 gene of S. cerevisiae is regulated in parallel with the hypoxic genes, but by a different mechanism. AB - The DAN1 gene is expressed under anaerobic conditions in yeast and completely repressed during aerobic growth. The function of the gene is unknown, and genetic disruption had no effect on fitness which could be detected, even upon prolonged anaerobic growth. Expression of DAN1 was constitutive in a heme-deficient strain, indicating that heme participates in repression. Expression was blocked by heme in anaerobic medium, suggesting that heme acts as a negative co-effector rather than through its metabolic functions, i.e., in the production of a co-effector. Expression of DAN1 was regulated in parallel with the hypoxic gene ANB1, showing identical kinetics of induction and dose response to heme. However, unlike ANB1, DAN1 is not regulated by the repressor of the hypoxic regulon, ROX1, as shown by observation of normal aerobic repression of DAN1 in a strain carrying a deletion of ROX1. These results indicate the existence of a parallel regulatory system which produces an identical response to oxygen by a different mechanism than that controlling the hypoxic regulon. PMID- 9224893 TI - The gene of the neural cell recognition molecule F11: conserved exon-intron arrangement in genes of neural members of the immunoglobulin superfamily. AB - The chicken neural glycoprotein F11 is a cell recognition molecule implicated in neurohistogenesis, in particular in the context of neurite outgrowth and fasciculation. F11 is a glycosyl-phosphatidylinositol-linked member of the immunoglobulin superfamily that is also termed contactin or F3 in humans and rodents, respectively. In this study, we report the complete structure of the F11 gene. It is composed of 23 exons distributed over more than 100 kb of genomic DNA and each of the ten domains of the F11 protein is encoded by two exons. The sizes of the introns vary by two orders of magnitude ranging from 150 bp to more than 15 kb. All interdomain introns are in phase one, i.e. are inserted after the first nucleotide of a codon, being consistent with assembly of a F11 progenitor gene via exon shuffling. The intradomain introns are localized at variable sites within the domains and have different intron phases. This study reveals a remarkable similarity of the F11 gene with the gene of axonin-1, a related neural immunoglobulin superfamily member which is also implicated in neurite outgrowth and fasciculation. The intron positions with respect to the protein domain organization are found to be identical, strongly suggesting that both genes are derived from a common ancestor that already had this exon-intron structure. PMID- 9224892 TI - Analyses of APG13 gene involved in autophagy in yeast, Saccharomyces cerevisiae. AB - We have isolated 14 apg mutants defective in autophagy in yeast Saccharomyces cerevisiae (Tsukada and Ohsumi, 1993). Among them, APG1 encodes a novel Ser/Thr protein kinase whose kinase activity is essential for autophagy. In the course of searching for genes that genetically interact with APG1, we found that overexpression of APG1 under control of the GAL1 promoter suppressed the autophagy-defective phenotype of apg13-1 mutant. Cloning and sequencing analysis showed that the APG13 gene encodes a novel hydrophilic protein of 738 amino acid residues. APG13 gene is constitutively expressed bot not starvation-inducible. Though dispensable for cell proliferation, APG13 is important for maintenance of cell viability under starvation conditions. apg13 disruptants were defective in autophagy like apg13-1 mutants. Morphological and biochemical investigation showed that a defect in autophagy of delta apg13 was also suppressed by APG1 overexpression. These results imply genetic interaction between APG1 and APG13. PMID- 9224894 TI - Characterization of the avian Ich-1 cDNA and expression of Ich-1L mRNA in the hen ovary. AB - We have sequenced the chicken interleukin-1beta-converting enzyme (ICE) and ced-3 homolog (Ich-1) cDNA, and evaluated Ich-1 mRNA expression in the hen ovary during follicle development. While two alternatively spliced forms of Ich-1, Ich-1L and Ich-1S, were amplified by PCR from an embryonic chicken cDNA library, only the Ich-1L form was found to be expressed in adult ovarian granulosa and theca tissues. The deduced amino acid (aa) sequence of ICH-1L is 70.8% identical to human ICH-1L and contains the conserved QACRG peptide active catalytic sequence characteristic of many ICE-related family of cysteine proteases. PMID- 9224896 TI - A ribosomal RNA operon from Pseudomonas stutzeri Zobell. AB - A ribosomal RNA operon from the marine bacterium, Pseudomonas stutzeri Zobell, was cloned and characterized by Southern hybridization and sequence analysis. The 16S rRNA, 23S rRNA, 5S rRNA and 2 tRNA genes (alanine and isoleucine) were identified by homology with sequences in GenBank. The rRNA gene exhibited typical eubacterial organization (16S-tRNAs-23S-5S). A putative ribosomal promoter and anti-terminator regions were also identified and described. Significant differences in spacing of the anti-terminator regulatory elements were observed between P. stutzeri Zobell and Escherichia coli. PMID- 9224895 TI - Sequence and promoter regulation of the PCK1 gene encoding phosphoenolpyruvate carboxykinase of the fungal pathogen Candida albicans. AB - The PCK1 gene encoding PEP carboxykinase (Pck1) of the fungal pathogen Candida albicans was isolated and sequenced. The deduced Pck1 protein has high homology to ATP-dependent Pck1 proteins in other species, especially to Pck1 of Saccharomyces cerevisiae (70% homology), but not to GTP-dependent Pck1 proteins. PCK1 transcript levels were efficiently repressed by glucose and derepressed (induced) on gluconeogenetic carbon sources. PCK1 regulation occurs on the level of transcription, as demonstrated by a fusion of the PCK1 promoter to the LAC4 reporter gene, yielding derepressed/repressed expression ratios of > 100. Homologous sequences in the PCK1 promoters of C. albicans and S. cerevisiae were identified. The PCK1 promoter may be useful to efficiently regulate expression and thereby test the function of genes in C. albicans. PMID- 9224897 TI - Apg1p, a novel protein kinase required for the autophagic process in Saccharomyces cerevisiae. AB - Autophagic protein degradation includes bulk protein turnover with dynamic membrane reorganization, in which formation of novel organelles autophagosomes play key roles. We have shown that Saccharomyces cerevisiae performs the autophagy in the vacuole, a lytic compartment of yeast, in response to various kinds of nutrient starvation. Here we show that the APG1 gene, involved in the autophagic process in yeast, encodes a novel type of Ser/Thr protein kinase. Our results provide direct evidence for involvement of protein phosphorylation in regulation of the autophagic process. We found overall homology of Apglp with C. elegans Unc-51 protein, suggesting that homologous molecular mechanisms, conserved from unicellular to multicellular organisms, are involved in dynamic membrane flow. PMID- 9224898 TI - Molecular analysis of the Rhodobacter capsulatus chaperone dnaKJ operon: purification and characterization of DnaK. AB - In Rhodobacter capsulatus (Rbc), the participation of DnaK in the synthesis of light harvesting antenna complex I (LHI) has been recently inferred from the finding that the amount of LHI alpha- and beta-polypeptides synthesized in an in vitro translation system was strongly reduced when DnaK was depleted. In the present work, a DnaK protein was isolated from Rbc and biochemically characterized. The N-terminus of the protein was sequenced and a corresponding oligo was used as probe in order to clone the gene coding for DnaK. The dnaK gene was located in an operon (dnaKJ) with two open reading frames, which code for DnaK and DnaJ, respectively. A promoter element corresponding to the consensus sequence of the atypical heat shock (HS) promoter of several alpha-purple proteobacteria was identified. Northern blot analysis indicated that dnaK and dnaJ belong to the same transcriptional unit; there were two transcripts, one comprising both the dnaK and dnaJ genes and a second with only dnaK. Primer extension analysis revealed that under both chemotrophic and phototrophic conditions transcription was initiated from the same position before and after HS. The promoter activity was studied under different growth conditions with a dnaK-lacZ fusion under the control of the dnaKJ promoter. The present work opens up the possibility to study the specific role of DnaK in the assembly of photosynthetic apparatus proteins. PMID- 9224899 TI - Cloning and expression of a chicken alpha-amylase gene. AB - We have isolated and sequenced a genomic clone for a pancreatic alpha-amylase gene (amy) of the chicken (Gallus gallus). The gene is interrupted by nine introns, spans over 4 kb, and encodes a protein (AMY) of 512 aa that is 83% identical to the human pancreatic alpha-amylase enzyme. Southern blot analysis of chicken DNA revealed two distinct pancreatic amy loci. In addition, we have generated a cDNA from chicken pancreatic RNA corresponding to the coding sequence of the genomic clone. The cDNA was inserted into a yeast expression vector, and the resulting construct used to transform Saccharomyces cerevisiae cells. Transformed yeast cells synthesized and secreted active AMY enzyme, and the gel migration pattern of the alpha-amylase produced by the yeast cells was identical to that of the native chicken enzyme. PMID- 9224900 TI - Single-column purification of free recombinant proteins using a self-cleavable affinity tag derived from a protein splicing element. AB - A novel protein purification system has been developed which enables purification of free recombinant proteins in a single chromatographic step. The system utilizes a modified protein splicing element (intein) from Saccharomyces cerevisiae (Sce VMA intein) in conjunction with a chitin-binding domain (CBD) from Bacillus circulans as an affinity tag. The concept is based on the observation that the modified Sce VMA intein can be induced to undergo a self cleavage reaction at its N-terminal peptide linkage by 1,4-dithiothreitol (DTT), beta-mercaptoethanol (beta-ME) or cysteine at low temperatures and over a broad pH range. A target protein is cloned in-frame with the N-terminus of the intein CBD fusion, and the stable fusion protein is purified by adsorption onto a chitin column. The immobilized fusion protein is then induced to undergo self-cleavage under mild conditions, resulting in the release of the target protein while the intein-CBD fusion remains bound to the column. No exogenous proteolytic cleavage is needed. Furthermore, using this procedure, the purified free target protein can be specifically labeled at its C-terminus. PMID- 9224901 TI - Cloning of Drosophila MCM homologs and analysis of their requirement during embryogenesis. AB - MCM (minichromosome maintenance) gene family of Saccharomyces cerevisiae encodes essential DNA replication factors that participate in the initiation of DNA replication. In addition, their localization to the nucleus in a mitosis dependent manner fueled the hypothesis that MCMs also act to couple DNA replication to mitosis. We report the identification of a Drosophila gene family with extensive sequence identity to the MCM genes. Results from antibody injection experiments suggest that MCMs play an essential role in DNA replication during embryogenesis. Evolutionary conservation of MCM sequences and function in Drosophila could potentially facilitate studies of how initiation of DNA replication is regulated and coupled to mitosis during metazoan development. PMID- 9224902 TI - Identification of a new member (ZNF183) of the Ring finger gene family in Xq24 25. AB - Four genes were mapped to the Xq24-25 region by searching the EST and the non redundant database with short tracts of genomic sequences. These were random STSs present in the STS database or sequences derived from CpG islands (EagI-based STSs). One of the four matches corresponded to the full length transcript from the intronless glutamate dehydrogenase gene. The second was the human homolog of the bovine NADH ubiquinone oxidoreductase MWFE subunit gene (GDB symbol: NDUFA1). The other two, ZNF183 and ITBA4, were novel genes whose function cannot directly be inferred from their sequence analysis. However, a known motif, the C3HC4 Ring finger domain, shared by various tumor suppressors, DNA repair genes and cytokine receptor-associated molecules, is present at the C terminus of the ubiquitously expressed ZNF183 gene. ITBA4 is expressed at various levels in different tissues and is alternatively processed in brain. Similarity search did not detect any significant match in databases. These results, together with others previously reported by our laboratory, suggest that comparison of genomic and transcribed sequences which are continuously accumulating in databases, can provide 'virtual' mapping of a substantial number of ESTs to the specific genomic region which the STSs have been derived from. PMID- 9224903 TI - Serotonin syndrome and drug combinations: focus on MAOI and RIMA. AB - Serotonin syndrome is a potentially life-threatening complication of psychopharmacological drug therapy. The syndrome is produced most often by the concurrent use of two or more drugs that increase brainstem serotonin activity and is often unrecognized because of the varied and nonspecific nature of its symptomatology. Serotonin syndrome is characterized by alterations in cognition, behavior, autonomic nervous system function and neuromuscular activity. The purpose of this study was to investigate the possibility that any serotomimetic substance alone or in combination may give rise to serotonin syndrome, that this condition is not confined to the use of newly introduced substances, and that the newer reversible inhibitors of monoamine oxidase type A (RIMAs) are at decreased risk for this phenomenon than older, classical (irreversible) monoamine oxidase inhibitors (MAOI). This is a hypothesis-generating study based on a review of all published cases of adverse effects arising in patients receiving serotomimetic substances or combinations. A wide range of substances were involved in 226 cases published worldwide since 1950 where there was any use of single or combined serotomimetic treatments. Of the 226 cases, 105 fulfilled the Sternbach criteria for serotonin syndrome. Some classes of drugs and individual substances were more commonly represented. This may arise from product utilization patterns or from the specific properties of the individual products. However, moclobemide, a RIMA, was represented in only 9 of the 226 published cases and 3 of the 105 defined serotonin syndromes, either in multi-drug combinations and/or in mixed drug overdose. One explanation for the small number of cases involving RIMAs could be the reversibility of these new products. In addition the small number of reports on moclobemide could be an effect of its short availability in routine use during the period of the literature review. We conclude that a spectrum of serotonergic hyperactivity, through to a defined serotonin syndrome, may arise from the use or combination of any serotomimetic substance, as this is a consequence of the mechanism involved, rather than the use of any specific product such as the new antidepressants. We further conclude that this condition is not confined to the use of newly introduced substances and that the newer reversible inhibitors of monoamine oxidase type A (RIMAs) may be at decreased risk for this phenomenon than older, classical (irreversible) (MAOI). Given that a spectrum of serotonergic hyperactivity was observed, this analysis prompts redefinition of the currently accepted criteria for serotonin syndrome. PMID- 9224904 TI - Hippocampal pyramidal cell disarray correlates negatively to cell number: implications for the pathogenesis of schizophrenia. AB - Brains from patients with therapy-refractory schizophrenia were examined with respect to pyramidal cell disarray in the hippocampus, a finding reported in some studies, but not confirmed in others. A significantly higher number of disarrayed cells was seen in the brains of the schizophrenic patients in all subfields of the Cornu Ammonis (CA) investigated. Compared with controls the schizophrenic patients also had significantly fewer pyramidal cells in the observed areas of CA1-CA3, but not in CA4. There were no signs of gliosis. Finally, there was a significant negative correlation between the number of disarrayed cells and the total number of pyramidal cells in CA1-3 in the group of probands and controls taken together, a finding interpreted as lending support to the idea of a prenatal migratory disturbance in the pathogenesis of schizophrenic syndromes. PMID- 9224905 TI - Functional EEG mapping and SPECT in detoxified male alcoholics. AB - Fifteen alcoholics diagnosed according to DSM-III-R, who were detoxified for at least 2 weeks and showed no clinical withdrawal signs, were investigated with 16 channel EEG mapping during resting, manumotor and music perception conditions, and were compared with 13 control persons. Single photon emission computed tomography (SPECT) using hexa-methyl-propilene-amine-oxime (HMPAO) labeled with 99m-technetium (99mTc) as tracer was performed separately (in patients only) and submitted to semiquantitative region of interest (ROI) analysis in 2 slices, 6 and 10 cm above canthomeatal line, respectively. Resting EEG showed increased power values in fast beta frequency band for the detoxified alcoholics. On cortical stimulation, patients showed signs of pathological EEG reactivity. Correlations of EEG parameters to cerebral blood flow (CBF) values (patients only) yielded coefficients around zero for all frequency bands (signs of uncoupling). All findings point to organic brain dysfunctions in these patients which extend beyond the period of withdrawal. PMID- 9224906 TI - Predictors of clinical and social outcomes after hospitalization in schizophrenia. AB - A prospective cohort study of schizophrenia was carried out in Sao Paulo, Brazil, in order to investigate clinical and social outcomes in schizophrenia and related psychoses after hospitalization. A sample of 124 individuals who were living in a defined catchment area and had been consecutively admitted to psychiatric hospitals in that area with clinical diagnoses of non-affective functional psychoses was followed up for 2 years. Assessments of clinical status and social adjustment at inclusion and at 2-year follow-up were carried out by means of standardized instruments, the PSE and the DAS. At the end of the follow-up period, 120 subjects (96.8%) were traced, and 103 (83.1%) were re-assessed. At the second assessment, the proportion of subjects with a nuclear syndrome of schizophrenia had halved (from 68.3% to 32.7%), 23.8% were symptom free and 60.2% showed at least one psychotic symptom. Presence of psychotic symptoms at follow up was best predicted by educational level (less than 4 years of formal education) and an initial DSM-III-R diagnosis of schizophrenia. The distribution of global social adjustment levels at 2-year follow-up was similar to that observed at the outset, with approximately one third of subjects showing good, one third showing intermediate and one third showing poor global social adjustment. Social disability was best predicted by longer duration of illness, worse social adjustment levels at inclusion and lower educational level. PMID- 9224907 TI - Use of potentially abusive psychotropic substances in psychiatric inpatients. AB - A series of 417 consecutively admitted psychiatric inpatients were studied with regard to their use of potentially abusive psychotropic substances in the last 3 months preceding admission. In all patients face-to-face interviews were performed; in 354 of them urine specimens could also be tested. Alcohol and benzodiazepines belonged to the most frequently used substances followed by cannabis, opiates and cocaine. Barbiturates, hallucinogens and amphetamine derivatives were only exceptionally reported. The most important finding of the study is that every fifth patient regularly used "hard" drugs (opiates and/or cocaine), every fourth patient illegal drugs and every third patient alcohol. Substances were found in 54% of all urine specimens; methadone, opiates and cocaine were hardly found alone. For the latter substances excellent agreement was found between interview reports and urine exams. Excluding patients diagnosed as substance-use disorders, there were no statistically significant differences between schizophrenic, affective, neurotic/stress/somatoform and other disorders with regard to the use of "hard" drugs and illegal drugs. Regular substance use correlated with much worse psychosocial adjustment. Substance use has to be explored and considered in every individual psychiatric inpatient. PMID- 9224909 TI - Mental illness in homeless women: an epidemiological study in Munich, Germany. AB - In an epidemiological survey of the prevalence of mental illness in homeless individuals in Munich, Germany, a probability sample of 32 homeless women were interviewed using a standardized diagnostic instrument (Diagnostic Interview Schedule for DSM-III diagnoses). Results point to very high prevalence rates of mental disorders among homeless women. The most frequent diagnostic groups were alcohol and drug abuse (lifetime prevalence rate 90.6%), affective disorders (50.0%), anxiety disorders (43.8%) and schizophrenia (21.9%). Prevalence rates are compared with a female household sample (Epidemiological Catchment Area Study in New Haven, Connecticut). All disorders tended to be more frequent in homeless women as compared with the household sample. Our results show the urgent need to provide medical and other assistance to homeless women. There is a need for adequate psychiatric care, supply with food and housing and the development of concepts for personal and vocational rehabilitation considering the specific needs of women. PMID- 9224910 TI - Malonic dialdehyde in the cerebrospinal fluid in intracranial hypertension. PMID- 9224908 TI - Serotonin-immune interactions in major depression: lower serum tryptophan as a marker of an immune-inflammatory response. AB - Serum total tryptophan and the five competing amino acids (CAA), i.e., valine, leucine, tyrosine, phenylalanine, and isoleucine were determined in 35 major depressed subjects of whom 27 with treatment resistant depression (TRD), and 15 normal controls. Twenty-five of the depressed subjects had repeated measurements of the amino acids both before and after antidepressive treatment. The following immune-inflammatory variables were assayed in the above subjects: serum zinc (Zn), total serum protein (TSP), albumin (Alb), transferrin (Tf), iron (Fe), high density lipoprotein cholesterol (HDL-C), number of peripheral blood leukocytes, and the CD4+/CD8+ T cell (T-helper/T-suppressor) ratio. Serum tryptophan and the tryptophan/CAA ratio were significantly lower in major depressed subjects than in normal controls. The tryptophan/CAA ratio was significantly lower in patients with TRD than in patients without TRD and normal controls. There were no significant alterations in any of the amino acids upon successful therapy. There were significant correlations between serum tryptophan and serum Zn, TSP, Alb, Tf, Fe, and HDL-C (all positive), and number of leukocytes and the CD4+/CD8+ T cell ratio (all negative). The tryptophan/CAA ratio was significantly and negatively related to the number of leukocytes and the CD4+/CD8+ T-cell ratio. The results suggest that (a) TRD is characterized by lower availability of serum tryptophan; (b) the availability of tryptophan may remain decreased despite clinical recovery; and (c) the lower availability of tryptophan is probably a marker of the immune-inflammatory response during major depression. PMID- 9224911 TI - Trigeminal neurinomas in infants. PMID- 9224912 TI - Choroid plexus papillomas of the III ventricle in infants. Report of three cases. AB - The III ventricle is an uncommon location for choroid plexus papilloma at any age. We describe three new cases of choroid plexus papillomas of the III ventricle (CPPs). All children were boys under 4 months of age and all presented with increased intracranial pressure, hydrocephalus and macrocephaly. The three were examined by preoperative computed tomography (CT) and ultrasonography. Two of them were investigated with magnetic resonance imaging (MRI). The first case was treated with a right corticofrontal transventricular approach and subtotal resection, so that he required a second operation through a transcallosal approach. In the other two cases a transcallosal approach was used. Two children needed permanent ventriculo-peritoneal shunts. The average follow-up of 4.3 years has revealed no neurological deficits in any case. The timing of and the need for shunting are major considerations. Clinical and imaging follow-up (CT and/or ultrasonography) are very helpful in controlling postoperative hydrocephalus and subdural effusion, avoiding unnecessary shunting in many cases. The operative approaches, transcortical and transcallosal, are discussed. PMID- 9224913 TI - Basal interhemispheric supra- and/or infrachiasmal approaches via superomedial orbitotomy for hypothalamic lesions: preservation of hypothalamo-pituitary functions in combination treatment with radiosurgery. AB - Although several approaches to the hypothalamus have been used, none is able to give full views of the hypothalamus. The risk of permanent morbidity for hypothalamo-pituitary functions is still high, especially in patients with craniopharyngioma. Basal interhemispheric supra-chiasmal or infra-chiasmal approaches via superomedial orbitotomy were developed for better visualization of the hypothalamus. Operative techniques and results, including combination treatment with radiosurgery, are reported. Twelve patients with tumors compressing the hypothalamus upward or extending into the III ventricle, or both, were operated on: 3 tumors were removed totally, 6 tumors subtotally and 3 tumors partially. Six patients received radiosurgery for residual tumor. Four patients with hypopituitarism preoperatively required oral corticosteroids and thyroid hormones postoperatively. The basal interhemispheric approach via superomedial orbitotomy is useful for better visualization of the hypothalamus and preservation of hypothalamo-pituitary functions. PMID- 9224914 TI - Hemiconvulsion-hemiplegia-epilepsy syndrome. A clinical, electroencephalographic and neuroradiological study. AB - Six patients (4 boys and 2 girls) with hemiconvulsion-hemiplegia-epilepsy (HHE) syndrome are described. They had prolonged seizures, lasting from 30 min to 12 h, at ages 1-4 years. These took the form of hemiconvulsion in three of the children and generalized tonic-clonic seizures in the others, being preceded by hemifacial twitching or head and eye deviation in two. They were followed by hemiplegia, which cleared with time in five patients, apart from subtle pyramidal tract signs. One child had spastic quadriparesis, choreiform movements, contracture deformities and severe mental retardation following repeated status epilepticus. Subsequent epilepsy developed in five patients and was satisfactorily controlled with carbamazepine and/or phenobarbitone. Cerebral hemiatrophy was documented in all patients by cranial computed tomography and/or magnetic resonance imaging. Single photon emission computed tomography (done in 4 patients) showed ipsilateral hypoperfusion (of the damaged hemisphere). Electroencephalography showed ipsilateral slowing and low voltage of background activity. Epileptiform discharges were found on the ipsilateral side in two cases and the contralateral side (the undamaged hemisphere) in one. PMID- 9224915 TI - Isolated ophthalmological manifestations due to malfunction of a lumboperitoneal shunt: shortening of the spinal catheter in three pediatric patients. AB - A long-term follow-up study was conducted in 28 pediatric patients treated with lumboperitoneal shunting. Shunt malfunction was observed in 3 patients (10.6%) who presented with isolated ophthalmological abnormalities 3, 5, and 11 years after surgery. The mechanism of this phenomenon is considered to be very slowly developing malfunction secondary to shortening of the spinal catheter associated with child's growth. The importance of periodic funduscopic examination in patients treated with lumboperitoneal shunting in infancy is stressed. PMID- 9224916 TI - Surgical treatment supposed natural history of the tethered cord with occult spinal dysraphism. AB - We retrospectively evaluated the pre- and postoperative course of 34 tethered cord patients with occult spinal dysraphism in an attempt to infer the natural history of this disorder and to determine the effectiveness of the surgical treatment. There were 32 cases with lumbosacral lipoma and 2 with tight filum terminale. The age at surgery ranged from 1 month to 47 years old. Eight patients, aged 1 month to 4 years old, were asymptomatic; 26 had neurogenic bladder (26 cases) or motor problems affecting the legs (8 cases). None of the patients older than 5 years of age were asymptomatic. Untethering of the spinal cord was performed in all cases. The postoperative follow-up period ranged from 5 months to 11 years. During these periods, 7 (88%) of the 8 asymptomatic patients remained neurologically intact, 6 (23%) of the 26 symptomatic patients showed improved symptoms, and 15 patients (58%) remained unchanged. These results indicate that the neurological symptoms will appear progressively in the tethered cord patients, and that prophylactic surgery should be considered as early as possible. PMID- 9224918 TI - Cerebrospinal fluid hydrothorax caused by transdiaphragmatic migration of a ventriculoperitoneal catheter through the foramen of Bochdalek. AB - Cerebrospinal fluid hydrothorax is a very rare complication following ventriculoperitoneal shunting. The authors report a case of a 3-year-old girl who developed cerebrospinal fluid hydrothorax (caused by migration of the intra abdominal catheter through the right vertebrocostal trigone of Bochdalek, the one most unlikely to be congenitally patent) and respiratory distress. The patient was successfully treated with thoracocentesis and shunt revision. PMID- 9224917 TI - Antley-Bixler syndrome. Description of two new cases and a review of the literature. AB - Antley-Bixler syndrome was first described in 1975, and to date 20 cases have been reported. In addition to brachycephaly, the syndrome is associated with midface hypoplasia, often with choanal stenosis or atresia, bilateral radiohumeral synostosis, multiple joint contractures, femoral bowing and long bone fractures, "pear-shaped" nose, dysplastic ears and, occasionally, urogenital or cardiac defects. Survival is closely linked to upper airway obstruction. This, in addition to craniosynostosis, also affects mental prognosis. The cluster of malformations and their severity are variable, and while numerous children have died early from respiratory distress, one third of them are alive and have had quite satisfactory development. With early and effective prevention of respiratory complications and early treatment of craniosynostosis, the overall prognosis can be favorable. The mode of inheritance is probably autosomal recessive, and midtrimester prenatal diagnosis is feasible. Genetic counseling depends on accurate prognostic and therapeutic data. We describe two new cases, a 4-year-old boy with unilateral coronal synostosis and radiohumeral synostosis on the same side and an 18-month-old girl with brachycephaly and imperforate anus. PMID- 9224919 TI - Hemidystonia secondary to carotid artery gunshot injury. AB - A 9-year-old boy was accidentally shot at close range with a pistol. The bullet entered through the left anterior neck and severed the left common carotid artery. Emergency surgery was performed with an end-to-end anastomosis. He recovered gradually from severe right-sided hemiparesis. CT scans demonstrated left parietal infarction. Within months he developed right hemidystonia, which progressed over the next few years. The movement disorder was refractory to medical therapy. MR scans showed a large demarcated defect in the left parietal lobe extending to the occipital lobe, to the insula and to the posterior ventral putamen. At age 18 the patient underwent a staged left-sided thalamotomy. The hemidystonia improved postoperatively but later partially recurred. PMID- 9224920 TI - Hemidystonia as presenting symptom of an optic glioma. AB - The first case is presented of an exophytic chiasmatic-hypothalamic glioma causing progressive hemidystonia in an 8-year-old boy, and all 24 reported cases of intraaxial cerebral tumours resulting in symptomatic hemidystonia are reviewed. PMID- 9224921 TI - Posttraumatic intradiploic arachnoid cyst of the posterior fossa. AB - We report the case of a 5-year-old girl with an enlarging suboccipital mass, a posttraumatic intraosseous arachnoid cyst. Diagnostic work-up revealed that the lesion consisted of an intradiploic arachnoid cyst and an extra-axial occipital pouch that communicated by way of an osseous and dural defect. Surgical repair was undertaken with good results. A search of the current literature has shown only seven previous reports of leptomeningeal cysts situated at the occipital bones, most of them the result of an antecedent skull fracture. A pathogenetic hypothesis is presented comparing the growth of arachnoid intraosseous cysts and the development of meningocencephaloceles. PMID- 9224922 TI - New developments in combinatorial libraries. PMID- 9224923 TI - Selectively infective phages (SIP). AB - We review here advances in the selectively infective phage (SIP) technology, a novel method for the in vivo selection of interacting protein-ligand pairs. A 'selectively infective phage' consists of two components, a filamentous phage particle made non-infective by replacing its N-terminal domains of gene3 protein (g3p) with a ligand-binding protein, and an 'adapter' molecule in which the ligand is linked to those N-terminal domains of g3p which are missing from the phage particle. Infectivity is restored when the displayed protein binds the ligand and thereby attaches the missing N-terminal domains of g3p to the phage particle. Phage propagation becomes strictly dependent on the protein-ligand interaction. This method shows promise both in the area of library screening and in the optimization of peptides or proteins. PMID- 9224924 TI - From the natural evolution to the genetic manipulation of the host-range of retroviruses. AB - The discovery that retroviruses share many features with retrotransposons led to the proposal that retroviruses evolved from cellular movable elements and became independent genetic entities, which started a parasitic life on their own. It has been further hypothesized that in the course of this evolutionary process, retrotransposons incorporated a cellular membrane glycoprotein gene into their genome, which finally became the envelope gene of the virus. Equipped with a error-prone polymerase and a high recombination rate during replication, the resulting genetic element has been equipped with a powerful machinery to change and modify itself, in particular the envelope gene, not only to escape the immune system, but also to change the host species. The HIV-1 epidemiology may serve as the best example of this still on-going evolutionary process. Since retroviruses insert their genome into that of the host cell as part of their life cycle, they have been extensively utilized as gene transfer vectors (retroviral vectors). Recently, experiments have been initiated to modify the envelope of retroviral vectors in the laboratory to alter the host range for specific gene transfer applications. This review highlights aspects of the natural evolution and the genetic modification of the retroviral envelope protein. Its implication on the safety of retroviral vectors in human gene therapy is also discussed. PMID- 9224925 TI - The nonstructural proteins of the hepatitis C virus: structure and functions. AB - The hepatitis C virus is the major causative agent of nonA-nonB hepatitis worldwide. Although this virus cannot be cultivated in cell culture, several of its features have been elucidated in the past few years. The viral genome is a single-stranded, 9.5kb long RNA molecule of positive polarity. The viral genome is translated into a single polyprotein of about 3000 amino acids. The virally encoded polyprotein undergoes proteolytic processing by a combination of cellular and viral proteolytic enzymes in order to yield all the mature viral gene products. The gene order of HCV has been determined to be C-E1-E2-p7-NS2-NS3-NS4A NS4B-NS5A-NS5B. The mature structural proteins, C, E1 and E2 have been shown to arise from the viral polyprotein via proteolytic processing by host signal peptidases. Conversely, generation of the mature nonstructural proteins relies on the activity of viral proteases. Thus, cleavage at the NS2/NS3 junction is accomplished by a metal-dependent autoprotease encoded within NS2 and the N terminus of NS3. The remaining cleavages downstream from this site are effected by a serine protease contained within the N-terminal region of NS3. Besides the protease domain, NS3 also contains an RNA helicase domain at its C-terminus. NS3 forms a heterodimeric complex with NS4A. The latter is a membrane protein that has been shown to act as a cofactor of the protease. Whereas the NS5B protein has been shown to be the viral RNA-dependent RNA polymerase, no function has yet been attributed to NS4B and NS5A. The latter is a cytoplasmic phosphoprotein and appears to be involved in mediating the resistance of the hepatitis C virus to the action of interferon. PMID- 9224926 TI - RNA-protein interactions in the regulation of coronavirus RNA replication and transcription. AB - Coronavirus, with a 31-kb RNA genome, replicates its own RNA and transcribes subgenomic mRNAs by complex mechanisms. Viral RNA synthesis is regulated by multiple RNA regions, which appear to interact either directly or indirectly. Multiple cellular proteins bind to these regions and may undergo additional protein-protein interactions. These findings suggest that coronavirus RNA synthesis is carried out on a ribonucleoprotein via a mechanism that involves both viral and cellular proteins associated with viral RNA, similar to DNA dependent RNA transcription. This mode of RNA synthesis may be applicable to most RNA viruses. PMID- 9224927 TI - The molecular anatomy of influenza virus RNA polymerase. AB - The genome of influenza virus is composed of eight RNA segments of negative polarity. The RNA-dependent RNA polymerase is associated with each viral RNA (vRNA) segment and in virus-infected cells, involved in both transcription, i.e. vRNA-directed synthesis of viral mRNA, and two step reactions of vRNA replication, i.e. vRNA-dependent synthesis of complementary RNA (cRNA) and cRNA dependent synthesis of vRNA. The RNA polymerase is composed of three viral proteins, PB1, PB2 and PA. PB1 is the core subunit for not only the RNA synthesis but also the assembly of PB2 and PA into this multifunctional enzyme complex. PB1 alone is able to catalyze vRNA-dependent RNA synthesis, but PB2 is required for capped RNA-dependent transcription, both together forming the transcriptase. The third P protein, PA, and an as yet unidentified host factor(s) are involved in the conversion of RNA polymerase from transcriptase to replicase. The functional map is being made for both PB1 and PB2 proteins. PMID- 9224928 TI - Role of cellular kinases in the gene expression of nonsegmented negative strand RNA viruses. AB - Nonsegmented negative strand RNA viruses package an RNA-dependent RNA polymerase composed of two subunits, a large protein L and a phosphoprotein P, for transcription and replication of their genome RNAs. The RNA polymerase activity resides within the L protein, while the P protein acts as a transcription factor or transactivator of the polymerase. Since P protein is heavily phosphorylated and phosphorylation is known to regulate function of many viral as well as cellular proteins, the role of phosphorylation of P protein in the gene expression of this group of RNA viruses has recently been investigated. Through expression in bacteria the P protein was produced in large quantity in the nonphosphorylated form and involvement of cellular kinase(s) in its phosphorylation was studied. Casein kinase II and/or protein kinase C have been shown to play a critical role in the activation of P protein in transcription. These findings have opened up a new avenue for studying an important regulatory step in virus gene expression that may lead to the development of an effective antiviral agent. PMID- 9224929 TI - Identification of HCV core mimotopes: improved methods for the selection and use of disease-related phage-displayed peptides. AB - Disease-specific epitope discovery from random peptide libraries displayed on phage using sera from patients involves a number of screening steps with many immune and non-immune sera. To rapidly identify mimotopes of the human hepatitis C virus (HCV) core protein, we have used an anti-core human monoclonal antibody (mAb; B12.F8) as a probe in screening phage that were affinity-selected using a serum from an HCV infected patient. Three different positive phage were isolated displaying low or no homology with the natural antigen, but which still efficiently bound to the antigen binding site of the B12.F8 antibody. Testing the reactivity of these phage with forty-five sera from HCV infected patients showed that antibodies recognizing them are present in more than 80% of this population. These antibodies showed distinct fine specificity, as they bound the selected phage in a mutually exclusive fashion. Co-expression of two mimotopes in the same cells led to chimeric particles which were recognized by antibodies of different specificity. These data provide novel information on the potential use of the phage display technology for the characterization of antibody specificity as well as disease diagnosis and prevention. PMID- 9224930 TI - A new immunohistochemical methodology for the specific detection of MDR1-P glycoprotein in human tissues based on phage-displayed peptides mimicking the MM4.17 epitope. AB - It is not rare that controversial indications about the presence or the expression level of multidrug-resistant (MDR) proteins come out from different laboratories upon examination of identical tumor specimens. Distinct aspects, including the use of weakly discriminating monoclonal antibodies (MAbs) and/or unsuitable techniques and procedures, contribute in generating differences in the MDR phenotype evaluation of cancer cells. In this regard we describe here an innovative immunohistochemical approach for the determination of P-glycoprotein expression in cells and tissues. The method is based on the ability of phage displayed peptides to mimic antibody epitopes. For this purpose we utilized the phage clone #55, which was affinity-purified from a phage-displayed random peptide library using the MAb MM4.17 (specific for MDR1-P-glycoprotein) as previously described. This clone has been chosen since it clearly and undoubtedly reacts with its cognate MAb, as was determined by ELISA and dot blot tests. Inhibition of the MAb MM4.17 binding to MDR1-P-glycoprotein-expressing cells could be performed by adding a calibrated concentration of phage clone #55 particles, which mimic MDR1-P-glycoprotein antigen. This methodology can eliminate misleading interpretations concerning the presence and expression level of MDR1-P-glycoprotein and might well contribute in routine clinical determinations of MDR in tumor specimens, thus contributing to our understanding of the basis of the mechanisms of tumor cell resistance to drugs. PMID- 9224931 TI - Assaying phage-borne peptides by phage capture on fibrinogen or streptavidin. AB - There is no simple and efficient method for assaying phage isolated from libraries without having to resort to PEG purification of the phage, or to the biotinylation or other labelling of the target molecule. We report here a method for producing 'bifunctional' phage that express two types of peptide; one peptide, fused to pVIII, will bind to immobilized fibrinogen, allowing capture of the phage out of culture supernatants; this allows the other peptide, fused to pIII or pVIII to be assayed by simple ELISA. This system has also been developed for the capture of phage bearing a streptavidin-binding peptide. The bifunctional phage are produced by bacterial cells bearing a plasmid that expresses pVIII fused either to the fibrinogen-binding peptide or to the streptavidin-binding one. Thus, when these cells are infected with a phage clone or pool to be assayed, phage will be produced whose 'capture-peptide' is produced from the plasmid and whose 'assay-peptide' is produced from the phage genome. We show here that, by this method, bifunctional phage can be produced that will bind to immobilized streptavidin or fibrinogen. PMID- 9224932 TI - Construction, exploitation and evolution of a new peptide library displayed at high density by fusion to the major coat protein of filamentous phage. AB - The amino-terminus of the major coat protein (PVIII) of filamentous phage can be extended, up to 6-7 residues, without interfering with the phage life cycle. We have constructed a library of approximately ten millions different phage each displaying a different octapeptide joined to the amino-terminus of the 2700 copies of PVIII. Most of the resulting clones are able to produce infective particles. This molecular repertoire constituted by the periodic regular decoration of the phage filament surface, can be utilized to search elements that bind proteins or relatively small organic molecules like the textile dye Cibacron blue. By sequential growth cycles we have performed a library evolution experiment to select phage clones that have a growth advantage in the absence of any requirement for binding a specific target. The consensus of the best growers reveals a Pro rich sequence with large hydrophobic residues at position 7 and Asn at position 1 of the random peptide insert. We propose that the assembly secretion process is favoured in phages displaying this family of peptides since they fit the groove between two adjacent PVIII subunits by making advantageous molecular contacts on the phage surface. PMID- 9224933 TI - Accessibility of peptides displayed on filamentous bacteriophage virions: susceptibility to proteinases. AB - The genome of the filamentous bacteriophage fd has been engineered so that small peptides can be inserted into the exposed N-terminal segment of pVIII, the major protein of the virus capsid. Most small peptides can be displayed on all 2700 copies of pVIII (a recombinant virion), but larger peptides can be displayed only in virions in which modified and wild-type proteins are intermingled (hybrid virions). Peptides displayed in this way are highly immunogenic and capable of interacting with receptors and other ligands. The physical accessibility of the displayed peptides was tested by examining their susceptibility to digestion with proteinases. Potential cleavage sites in peptides displayed on recombinant or hybrid virions were in general found to be accessible to trypsin and chymotrypsin; and the density of incorporation of peptides in the virion had no effect on the susceptibility to cleavage. However, peptide bonds towards the C terminal end of an insert, located approximately 47 residues or fewer from the C terminus of the coat protein, were protected from digestion, presumably because of their proximity to the bulk viral surface. These results have important implications for the design and optimization of peptide display systems using filamentous bacteriophages. PMID- 9224934 TI - Using molecular repertoires to identify high-affinity peptide ligands of the WW domain of human and mouse YAP. AB - The WW domain is a globular protein domain that is involved in mediating protein protein interaction and that ultimately participates in various intracellular signaling events. The domain binds to polyproline ligands containing the xPPxY consensus (where x signifies any amino acid, P is proline and Y is tyrosine). One of the first WW domain-ligand links that was characterized in vitro was the WW domain of Yes-Associated Protein (YAP) and its WBP-1 ligand. To further characterize this molecular interaction, we used two independent approaches, both of which focused on the mutational analysis of the WBP-1 ligand. We screened repertoires of synthetic decamer peptides containing the xPPxY core of WBP-1 in which all ten positions were sequentially replaced with the remaining amino acids. In addition, we screened decamer repertoires with all permutations of the amino acids which individually increased the binding to the WW domain of YAP, as compared to the wild type. In a parallel approach, we used a phage-displayed combinatorial peptide library biased for the presence of two consecutive prolines to study ligand preferences for the WW domain of YAP. Interestingly, these two lines of investigation converged and yielded the core sequence PPPPYP, which is preferred by the YAP-WW domain. This sequence was found within the p53 (tumor suppressor) binding protein-2, a probable cognate or alternative ligand interacting with YAP. PMID- 9224935 TI - Theoretical and experimental selection parameters for HBV-directed antisense RNA are related to increased RNA-RNA annealing. AB - Annealing kinetics of antisense species against two different target regions of the hepatitis B virus (HBV) were measured by kinetic in vitro selection. Individual association rates were related to energies calculated for local sequence segments and predicted structures of the complete pregenomic target RNA. A relationship between the presence of external loops and joint sequences with fast pairing was observed whereas internal loops did not favor fast RNA-RNA annealing. The findings were used to predict a fast-annealing HBV-directed antisense oligodeoxyribonucleotide that turned out to pair with its target RNA at an association rate constant of k=9.2 x 10(4) M(-1) s(-1), which is substantially faster than the annealing rates of artificial antisense RNA so far included in in vitro selection assays. PMID- 9224936 TI - Overproduction of Sac7d and Sac7e reveals only Sac7e to be a DNA-binding protein with ribonuclease activity from the extremophilic archaeon Sulfolobus acidocaldarius. AB - Genomic DNA from Sulfolobus acidocaldarius was screened using a degenerate oligodeoxyribonucleotide, derived from the sequence of 16 N-terminal amino acids from SaRD protein. SaRD protein was previously isolated in our laboratory and identified as a protein from S. acidocaldarius exhibiting ribonuclease activity as well as DNA-binding properties. On the basis of Southern hybridization analysis two genes from S. acidocaldarius have been cloned, sequenced and overproduced in Escherichia coli. The deduced amino acid sequences revealed that one gene encodes Sac7d and the other one Sac7e; two small, previously described basic proteins from S. acidocaldarius, and furthermore the N-termini of Sac7e and SaRD are identical. Northern blot analysis demonstrated that the genes are transcribed separately. After expression of sac7d and sac7e genes in E. coli it was shown that only recombinant Sac7e protein exhibits RNase activity and is catalytically indistinguishable from SaRD protein. Western blot analysis using a polyclonal antiserum raised against purified SaRD protein further confirmed that Sac7e and SaRD are identical proteins endowed with RNase activity and DNA-binding properties. A new RNA cleavage mechanism has to be postulated for Sac7e since, in contrast to common RNases (e.g. RNase A and T1), no histidines are present in the amino acid sequence. Differences between the very closely related 7 kDa proteins from two Sulfolobus strains converting DNA-binding proteins into RNases are pointed out and discussed, whereas substitutions of Glu by Gln (S. solfataricus) or by Lys (S. acidocaldarius) seem to be crucial. PMID- 9224937 TI - Ribonuclease T1 is active when both catalytic histidines are replaced by aspartate. AB - The enzymatic activity of many ribonucleases (RNases) depends on two histidines, as in RNase A, or one histidine and/or glutamate, as in microbial RNases belonging to the T1 family. In RNase T1, substitution of either one or both of the two histidines at positions 40 and 92 by a variety of other amino acids reduces the activity more than 100-fold. However, the double variant His40- >Asp/His92-->Asp retains a significant residual enzymatic activity towards RNA and guanylyl-3',5'-cytidine, indicating that a pair of substituted side chains in these positions, both with acid functionality, can confer enzymatic activity. It was shown that the substitution of histidine with glutamate in the variant His40- >Glu yields an enzyme with drastically reduced activity and leads to inactivation in the His92-->Glu, His40-->Glu/His92-->Glu variants. For the variants where histidine is substituted with aspartate we found measurable activity from 1% (His40-->Asp) to 6% (His40-->Glu/His92-->Asp) towards RNA. PMID- 9224938 TI - Inducible expression of the beta 1 subunit of the sodium pump. AB - The Na,K-ATPase, or sodium pump, a ubiquitous transmembrane enzyme in higher eukaryotes, consists of an alpha and a beta subunit. Here we investigate the expression pattern of the two beta isotypes in mouse B cell lines. Neither primary cells nor cell lines express beta 2. Abelson virus-transformed pre-B cells express beta 1, while B lymphomas and plasmacytomas do not. Thus, beta 1 expression in transformed cells follows that of their untransformed counterparts. Some subclones of pre-B cell line 70Z/3 express beta 1, and others do not, but lipopolysaccharide induces the beta 1-negative cells to become beta 1-positive. PMID- 9224939 TI - A novel SR-related protein specifically interacts with the carboxy-terminal domain (CTD) of RNA polymerase II through a conserved interaction domain. AB - The largest subunit of the RNA polymerase II (pol II) contains at the carboxy terminus a peculiar repetitive sequence that consists of 52 tandem repeats of the consensus motif Tyr-Ser-Pro-Thr-Ser-Pro-Ser, referred to as the C-terminal domain (CTD). Upon transcriptional initiation/promoter clearance, the CTD becomes extensively phosphorylated and apparently remains so during elongation. While the underphosphorylated CTD plays a role in transcriptional initiation, recent evidence couples the highly phosphorylated CTD to RNA processing, namely polyadenylation and splicing. Using a yeast two-hybrid screen, we have selected for human proteins that interact with the CTD of RNA polymerase II. The CTD-GAL fusion protein used as a bait is highly phosphorylated in yeast and, accordingly, we did not isolate proteins implicated in transcriptional regulation but rather proteins with possible roles in RNA splicing. One major cDNA clone isolated this way encodes SRrp129/CASP11, a protein that contains a conserved CTD-interaction domain at the C-terminus and an internal serine-arginine rich domain (SR domain). Proteins of the SR family have been implicated in RNA splicing, notably in the regulation of alternative splicing. Thus we consider it likely that SRrp129 is an auxiliary splice factor. We also improved our method to quickly map domains involved in protein-protein interaction (Stagljar et al., 1996, BioTechniques 21, 430-432). Instead of using sonication for the production of a random DNA fragment library, we took advantage of the fact that DNAse I in the presence of manganese (II) produces double strand rather than single strand DNA breaks. The DNA fragment library of the SRrp129 clone was then used in the yeast two-hybrid system to identify the 100-amino acid domain that interacts with the CTD of RNA polymerase II. PMID- 9224940 TI - Complex segregation analysis of facial clefting in Chile. AB - Nonsyndromic cleft lip with or without cleft palate (CL/P) has an incidence of 1.5 per 1,000 live births in Chile, with 1.7 per 1,000 in males and 1.3 per 1,000 in females, which is nearly the same as the level found in Asian populations. The high rate of occurrence of CL/P in Chile is probably due to the presence of Amerindian genes in Chilean populations. Using the computer program PAP, a complex segregation analysis of CL/P was conducted for 67 multigeneration pedigrees from Chile, each ascertained from one affected proband. These pedigrees yielded 162 affected individuals and over 898 family members who were included in the analysis. The most parsimonious model of transmission indicated the presence of an autosomal dominant gene with reduced (20-25%) penetrance. PMID- 9224941 TI - Cleft lip and palate etiology and its meaning in early 20th century England: Galton/Pearson vs. Bateson; polygenically poor protoplasm vs. Mendelism. AB - At the outset of the 20th century in England there arose a venomous dispute between Mendelian geneticists such as Bateson and anti-Mendelian biometricians such as Pearson over the genetic etiology of such "physical deformities" as cleft lip and palate. To Pearson et al., such traits were an expression of physical and racial degeneracy which could be traced to polygenically poor protoplasm. To Bateson et al., such traits were Mendelian unit characters whose segregation could be seen in carefully constructed family pedigrees. Bateson dismissed the work of the anti-Mendelians as "unsound in construction" and predicted such thinking would inevitably lead to "brutal" control of those the larger society deemed unfit. History proved Bateson astutely prescient. PMID- 9224942 TI - Mandibular movements during elevation and fusion of palatal shelves evaluated from the course of Meckel's cartilage. AB - The purpose of the present study was to investigate whether the course of Meckel's cartilage could reveal the mandibular movements during the elevation and fusion of the soft tissue palatal shelves. Histological sections, cut serially in the horizontal plane from 64 human mandibles, 16-104 mm CRL, were analyzed. The course of the anterior and medial part of Meckel's cartilage changed markedly during the three palatal stages, i.e., before, during, and after palate formation. The medial part changed during these stages from a straight course through a curled, S-shaped course to a crochet-hook-shaped course. The anterior part of Meckel's cartilage developed from a separation in the symphysis menti region to a fusion and later to a separation again. It is suggested that these changes in the course of Meckel's cartilage are due to different muscle activities. It is supposed that during the palatal developmental stages the activity of the geniohyoid and genioglossus muscles caused the mandibular retraction and the widening of the angulation between the bilateral hemimandibles. The S-shape of Meckel's cartilage is a result of these movements. Later mandibular proclination and narrowing of the bilateral hemimandibles resulted in an anterior mechanical fusion of Meckel's cartilage due to the activity of the mylohyoid muscle. This stage is followed by a retraction and re widening of the angulation between the bilateral bony components, which disrupts the fusion of Meckel's cartilage in the symphysis menti region. Thus, the course of Meckel's cartilage revealed the mandibular movements in the sagittal and transverse planes during palate formation. PMID- 9224943 TI - Soft tissue facial morphology related to headform: a three-dimensional quantitative analysis in childhood. AB - The object of this investigation was to determine whether children of the same age with different headforms differ in their three-dimensional soft-tissue facial characteristics. The three-dimensional coordinates of 22 standardized facial landmarks were automatically collected in a sample of 70 boys and 71 girls age 11 to 13 years attending a junior high school. From the collected landmarks, several three-dimensional facial angles, linear distances, linear distance ratios, and volumes were calculated. For each subject the cephalic index (maximal head breadth/ maximal head length x 100) was computed and three groups of measurements for each sex were obtained (dolicho-, meso- and brachycephalic). A two-way factorial analysis of variance compared the effects of sex and headform, and the interaction sex x headform. On average, boys had significantly (P < or = 0.05) longer and wider faces than girls, with a larger lower third facial volume relative to middle third facial volume. A significant (P < or = 0.05) effect of headform over facial morphology was found for all angles with a prevalent axial orientation. Conversely, no effect was demonstrated for angles with a sagittal orientation, nor for any other considered parameters. For each sex, the dolichocephalic children had smaller values than the brachycephalic children (i.e., more convex faces in the left-right direction), while the mesocephalic children had intermediate values. No sex x headform interactions were found. Results confirm that a different headform (skull) is associated with a different three-dimensional facial morphology (combined effect of skull and soft tissues), but without size differences. PMID- 9224944 TI - Dietary consistency and craniofacial development related to masticatory function in minipigs. AB - Since the 1890s oral biological researchers have been interested in the idea that strenuous mastication of unprocessed food will stimulate proper oral-facial growth and occlusal relationships. Conversely, lack of such function due to consumption of refined food is one hypothesis among many for the etiology of malocclusion in industrialized humans. Adequately controlled experimental testing of the idea has been limited to rats. To investigate the "disuse" theory in a larger-bodied and more occlusally relevant animal model, we raised four Yucatan minipigs from weaning on hard diet (HD) and another four on softened but equivalent diet (SD). The animals were monitored for eight months, sacrificed, and then occlusal and osteometric data collected. Variations due to dietary regime are pervasive and not due to caries, periodontitis, or attrition differences. Whereas HD body weight is 10% greater than SD, the deep masseter is 25% greater, with similar disproportion in superficial masseter and temporalis weight. Facial prognathism, arch narrowness, tooth crowding/maleruption and posterior cranial tapering are markedly different in the two groups. A curious posterior torsional difference in the mandibular rami, as well as broadness and flatness of the mandibular symphysis, also occur in SD. We performed a Q-mode principal coordinates analysis of the 19 logged variables for the specimens, bootstrapping the variable list, to demonstrate a statistically significant (P < .01) overall pattern of dramatic differences. Having controlled other celebrated orthodontic etiologies (genetic background, respiratory mode, infectious degeneration and interproximal attrition), these results support the proposition that dietary consistency relates directly to human craniofacial growth. PMID- 9224945 TI - Molecular cloning and sequencing of the cDNA encoding the catalytic subunit of mouse glutamate-cysteine ligase. AB - Reverse transcription-polymerase chain reaction (RT-PCR) was used for the enzymatic synthesis of cDNA sequences encompassing the open reading frame for the catalytic subunit of mouse kidney glutamate-cysteine ligase (Glclc). Comparison of the mouse Glclc cDNA sequence and predicted protein sequence with that of rat Glclc and human GLCLC revealed between 94.8% and 88.4% cDNA homology and 98.4% to 95% amino acid identity, respectively. PMID- 9224946 TI - Structure of the promoter for the rat Fas antigen gene. AB - The Fas antigen is a receptor protein transducing cell death signals. Binding of Fas ligands to Fas antigens provokes apoptosis in target cells. Here we report the structure of the promoter of the gene coding for rat Fas antigens. The major transcription start site, identified by the S1 nuclease protection assay, was situated 188 nucleotides upstream of the translational initiation site. The promoter activity was located in the region at the nucleotide position from -142 to -24. In this region we identified a consecutive sequence of NF-kappaB and NF IL6 consensus sequences, spanning from -142 to -122. PMID- 9224947 TI - Cloning and analysis of the dnaG gene encoding Pseudomonas putida DNA primase. AB - The dnaG gene coding for primase, a key enzyme in DNA replication, has been isolated from chromosomal DNA of the soil bacterium Pseudomonas putida. It maps within the putative MMS operon, between the rpsU and rpoD genes. Comparison of the deduced amino acid sequence of P. putida DnaG with sequences of other known bacterial primases reveals the presence of a possible regulatory region which would be unique to pseudomonads. The analysis of nucleotide sequence suggests that stable folding of the dnaG mRNA may significantly contribute to the low level of its expression within a cell. PMID- 9224948 TI - Cloning and sequence analysis of human calcyphosine complementary DNA. AB - Calcyphosine, initially identified as thyroid protein p24, is a calcium-binding protein containing four EF-hand domains. It was first cloned and characterized in the dog and corresponds to R2D5 antigen in rabbit. Using the canine calcyphosine cDNA sequence as a probe, we have isolated its human counterpart from a thyroid cDNA library. The two sequences display a high degree of conservation, both at nucleotide and deduced amino acid levels. Sequence comparison with other proteins showed that the closest homologue of calcyphosine is the crustacean CCBP-23 protein. Northern blot analysis revealed that calcyphosine messenger RNA is much less abundant in human than in canine thyrocytes. Western blot experiments indicated that the amount of protein is also dramatically reduced in man compared to dog. PMID- 9224949 TI - Sequence polymorphism and structural analysis of timothy grass pollen profilin allergen (Phl p 11). AB - Three cDNA clones encoding timothy grass pollen profilin (Phl p 11) were newly isolated. Comparison of the sequences of four cDNA clones, including a previously isolated clone, showed a low level of polymorphism. Isoelectrofocusing of highly purified timothy grass profilin indicated the existence of at least five isoforms. One recombinant profilin showed similar immunological properties to natural timothy grass profilin. Tertiary structure of Phleum pratense profilin was obtained by homology-based molecular modeling. PMID- 9224950 TI - The cloning and overexpression of a cruciform binding protein from Ustilago maydis. AB - The structural gene HMP1 encoding a cruciform DNA binding protein from Ustilago maydis has been cloned. Gene isolation was enabled by a polymerase chain reaction procedure using primers designed from amino acid sequence obtained from the purified protein. DNA sequence determination has revealed that the gene encodes a protein containing 98 amino acids with a calculated molecular weight of 10151. Comparison of the cDNA and genomic sequences indicated the presence of a single intron in the 5' coding region of the gene. The gene was over-expressed as a translational fusion with a hexahistidine leader sequence enabling affinity purification of the protein on an immobilized metal matrix. Protein isolated after over-expression exhibited cruciform binding activity, conforming earlier purified native protein results. Sequence analysis indicated that no HMG box was present and very little homology to other known cruciform binding proteins was found. It is plausible that HMP1 represents a novel class of proteins that recognize such secondary structures. PMID- 9224951 TI - Characterization and sequence of an additional 15-lipoxygenase transcript and of the human gene. AB - 15-lipoxygenase is a lipid-peroxidating enzyme that oxidizes fatty acids, such as those esterified to cellular membranes. It has been implicated in the oxidative modification of low-density lipoprotein and is thus thought to contribute to the development of atherosclerosis. The enzyme has also been shown to be specifically induced by interleukin-4 in human blood monocytes. Two 15-lipoxygenase hybridizing messages were detected in these cells; one (2.7 kb) corresponds to the previously isolated cDNA for 15-lipoxygenase, while the other (4 kb) was of unknown origin. We have isolated and characterized this 4 kb transcript. Our experiments show that it has 1.2 kb additional sequence in its 3' untranslated region, and that it is generated from genomic sequences through differential polyA site selection. We present studies to address the functional significance of the extended 3'UTR. Selection of an upstream polyadenylation signal results in production of the 2.7 kb transcript. In addition, we present here for the first time the cloning and sequence of the human 15-lipoxygenase gene, as well as the identification of regulatory elements in the promoter region of this gene. PMID- 9224952 TI - Complete cDNAs for CDC42 from chicken cochlea and mouse liver. AB - CDC42 is a member of the ras superfamily of small GTP-binding proteins that are related through the highly conserved GTP-binding domain and are involved in signal transduction pathways. Two full-length CDC42 cDNAs have been isolated: a 2148-bp chick cochlea cDNA and a 2063-bp mouse liver cDNA. Each encodes a CDC42 protein of 191 amino acids. The avian CDC42 protein differs from the mouse at only one amino acid residue, a Thr for a Ser at position 185. Both CDC42 proteins are more similar to the ubiquitous human isoform originally isolated from placenta than to the isoform isolated from fetal brain. Using a probe from the 3' UTR of the mouse liver CDC42 cDNA, we demonstrated that the mouse gene is expressed in all tissues examined. Southern blot analysis of a mouse inter specific backcross with this gene-specific probe identified at least three CDC42 like (Cdc42l) genes in the mouse genome. Cdc42l1 was mapped to distal mouse Chromosome 4, near Cappb1. Cdc42l2 mapped more proximal on Chromosome 4, whereas Cdc42l3 mapped to the X Chromosome. PMID- 9224954 TI - CpG island methylation and promoter usage in the parathyroid hormone-related protein gene of cultured lung cells. AB - Excessive production of a parathyroid hormone-related protein (PTHrP) by tumours commonly results in the syndrome of humoral hypercalcaemia of malignancy. We have investigated whether epigenetic changes play a role in over-expression of the PTHrP gene, using cultures lung cells as a model system. Study of the methylation status of CpG dinucleotides in the 5' region of the gene showed that in normal cells the CpG island was completely unmethylated. In the lung squamous cell carcinoma cell line, BEN, two-thirds of the CpG island was substantially methylated. RT-PCR analysis showed that this heavy methylation did not prevent expression of any of the three PTHrP gene promoters. This is a surprising finding, since methylation is usually associated with inhibition of gene activity. Methylation of the 5' non-coding region of the PTHrP gene may not play a role in the regulation of adjacent promoters. Alternatively, maintenance of a demethylated state in the 170 bp at the 3' end of the CpG island may be fundamental for the use of PTHrP promoters. PMID- 9224953 TI - Regulation of expression of the human heme oxygenase-1 gene in transfected chick embryo liver cell cultures. AB - Induction of heme oxygenase (HO) has been proposed as a protective cellular mechanism against oxidative damage. In previous work (Tyrrell et al., Carcinogenesis [1993] 14, 761-765), portions of the 5' promoter region of the human HO-1 gene linked to the reporter gene chloramphenicol acetyl transferase (CAT), had been transiently expressed in HeLa cells. To extend the study of human HO gene expression into primary liver cells, these reporter gene fusion constructs, containing 121 or 1416 base pairs of the untranscribed 5'-upstream sequences of the human HO-1 gene, were used along with pSV beta-Gal plasmid to dually transfect primary cultures of chick embryo liver cells (CELC). The transfected cells were treated with selected metals, heme, phorbol ester, and chemical agents that produce oxidative stress (H2O2 or sodium arsenite). Reporter gene activities were measured 18-20 h later. Our major findings are: (1) these HO CAT constructs were expressed in CELC; (2) unlike HeLa cells, the expression of CAT was detected in CELC without the need for the SV40 enhancer; (3) sodium arsenite and cobalt chloride induced the expression of the HO-CAT constructs whereas heme had no effect on or decreased CAT expression for all of the transfected constructs; (4) study of endogenous chick HO-1 gene expression in CELC showed that HO-1 responded to sodium arsenite treatment in a dose-dependent fashion, and the response was rapid and transient. We conclude that, in chick liver cell cultures, induction of the HO-1 gene by heme is fundamentally different from that produced by transition metals or sodium arsenite. Furthermore, the results suggest that expression of the HO-1 gene is highly conserved across species. PMID- 9224955 TI - Promoter of the Na,K-ATPase alpha3 subunit gene is composed of cis elements to which NF-Y and Sp1/Sp3 bind in rat cardiocytes. AB - Na,K-ATPase alpha subunit has three isoforms whose expression is regulated developmentally and hormonally. Na,K-ATPase alpha3 subunit gene (Atpla3) is expressed only in brain and neonatal heart in a rat. The purpose of this study is to analyze cis-acting elements and trans-acting factors regulating the transcription of Atpla3 in cultured neonatal rat cardiocytes. Transient transfection assays with Atpla3-luciferase chimeric construct and a series of 5' sequential deletion mutations revealed the existence of positive regulatory elements from -74 to -59 and from -59 to -39. A factor was identified to bind across -59 by gel retardation assay. Methylation interference and DNase I footprinting analyses revealed the binding region from -74 to -53 (positive regulatory element (PRE) 1). The binding factor was identified to be NF-Y by gel retardation assay using specific antibody. Gel retardation and methylation interference analyses revealed that factors bind to two other elements from -54 to -43 (PRE2) and from -25 to -13 (PRE3). The binding factors were identified to be Sp1/Sp3 using specific antibodies. The functions of above-mentioned three elements were examined by transient transfection assay with various combinations of mutations. They all regulated the transcription positively and a synergistic enhancement of it was observed. Roles of NF-Y in the transcriptional activation and synergy are discussed. PMID- 9224956 TI - Up-regulation of nuclear genes in response to inhibition of mitochondrial DNA expression in chicken cells. AB - Vertebrate cells depleted of (rho0) mitochondrial DNA (mtDNA) exhibited phenotypic traits that differed from the parental (rho+) cells. To isolate genes whose expression is associated with mtDNA depletion, we constructed cDNA libraries from mRNAs isolated from chicken rho+ cells transformed by the MC29 (v myc-containing) retrovirus and from rho0 cells developed by long-term exposure of the rho+ cells to ethidium bromide (EtdBr). Through subtractive hybridization procedures, three genes, elongation factor 1 alpha (EF- 1 alpha), beta-actin and v-myc were identified and found to be up-regulated in rho0 cells. In addition, Northern analysis demonstrated that the mRNA content for GAPDH was also elevated in rho0 cells. Run-on transcription assays and mRNA stability studies in the presence of actinomycin D indicated that elevated expression of these four genes depends, at least in part, upon increased rate of transcription. Other regulatory mechanisms contribute to the elevated expression of the transcripts in rho0 cells, as suggested by cycloheximide enhancement of the accumulation of the mRNAs for EF-1 alpha and beta-actin in rho0 cells, but not in parental rho+ cells. Moreover, inhibition of mtDNA replication and transcription by EtdBr and inhibition of translation on mitoribosomes by chloramphenicol also increased the expression of the four genes in parental rho+ cells, thus mimicking the situation in rho0 cells. These data suggest that information encoded within mtDNA participates in the regulation of nuclear genes in chicken cells. PMID- 9224957 TI - Herpesvirus proteases: targets for novel antiviral drugs. AB - Herpesvirus proteases have emerged as targets for the development of novel antiviral drugs. These enzymes, which are necessary for the replication of all herpesviruses, are serine proteases, but possess a unique structure as revealed by solution of the crystal structure of human cytomegalovirus protease. Many of the biochemical properties of these enzymes are now explained by the structure. Conventional serine protease inhibitors are not potent inhibitors of these enzymes and therefore the search for potent inhibitors possessing necessary features of an effective antiviral will require novel approaches. The three dimensional structure serves as a milestone for continued endeavors towards this goal. PMID- 9224958 TI - (S)-1-(3-hydroxy-2-phosphonylmethoxypropyl)cytosine (HPMPC) inhibits HIV-1 replication in epithelial cells, but not T-lymphocytes. AB - PMEA [9-(2-phosphonylmethoxyethyl)adenine] inhibited both HSV-1 and HIV-1 replication in MT-2 and HeLa-CD4 cells. (S)-1-[3-hydroxy-2 (phosphonylmethoxy)propyl]cytosine (HPMPC) inhibited both these viruses in the epithelioid HeLa-CD4 cells, but did not inhibit either virus in the T-lymphocytic MT-2 cells. PMEA and HPMPC are metabolized to their diphosphorylated forms within cells, which then inhibit viral polymerases. We therefore compared the metabolism of PMEA and HPMPC in MT-2 and HeLa CD4 cells. PMEApp formation was efficient in both the cell types, whereas HPMPCpp levels were approximately 3-10 fold lower in MT-2 cells, compared to HeLa-CD4 cells. These results indicate that HPMPC can inhibt HIV replication in the appropriate cell types, and show that differences in their metabolism cannot account entirely for the lack of antiviral efficacy of HPMPC in MT-2 cells. PMID- 9224959 TI - Rabies virus infection of IMR-32 human neuroblastoma cells and effect of neurochemical and other agents. AB - IMR-32 human neuroblastoma cells are a continuous nerve cell line expressing neuronal nicotinic acetylcholine receptors. These cells were found to be susceptible to infection by rabies virus (CVS strain). After infection, viral antigen accumulated in the cell body in puncta and larger masses and spread out into the processes until at 3-4 days the entire cell was filled with antigen and lysed. A variety of chemical agents including cholinergic agonists and antagonists were tested for ability to inhibit infection of IMR-32 cells in a fluorescent focus assay. Agents found to inhibit infection were antibodies against the viral glycoprotein, gangliosides, a synthetic peptide of the neurotoxin-binding site of Torpedo acetylcholine receptor alpha1 subunit, alpha bungarotoxin, and lysosomotropic agents. All other agents tested including other cholinergic ligands and synthetic peptides were not effective. Except for lysosomotropic agents, the agents which inhibited infection also inhibited attachment of virus to the cell surface. These results indicate that IMR-32 cells are a useful model in studying the interaction of a neurotropic virus with human neurons. The ability of alpha-bungarotoxin to inhibit infection suggests that neuronal alpha-bungarotoxin-binding receptors might serve as central nervous system receptors for rabies virus. PMID- 9224960 TI - A randomized, double-blind trial of parenteral low dose versus high dose interferon-beta in combination with cryotherapy for treatment of condyloma acuminatum. AB - Forty-nine subjects were enrolled in a study comparing two dosages of parenterally administered interferon (IFN)-beta in combination with cryotherapy for the treatment of anogenital warts. Subjects were randomized to receive subcutaneous injections of either 2 x 10(6) or 4 x 10(6) IU/m2 of IFN-beta (Biogen) three times a week for a total of 6 weeks. Cryotherapy was administered concomitantly by aerosolization of liquid nitrogen at 10-day intervals. Systemic side- effects were modest in intensity and included fever, chills, myalgia, and headaches (flu-like symptoms). During the first 2 weeks of therapy, they were more common in the high dose group than in the low dose group (P = 0.02). Using survival analysis, there was no significant difference between the two groups in rates of resolution of warts present at baseline (P = 0.62). However, the rate of new lesion formation during the study was significantly lower in the high dose group (P = 0.04). PMID- 9224961 TI - Pharmacokinetic study of the interaction between rifabutin and delavirdine mesylate in HIV-1 infected patients. AB - The oxidative metabolism of delavirdine, a non-nucleoside inhibitor of HIV-1 reverse transcriptase, is mediated in part by cytochrome P450 3A. The influence of rifabutin, an inducer of certain human cytochrome P450 isozymes, on the steady state pharmacokinetics of delavirdine was investigated in 12 HIV-positive patients with CD4 counts ranging from 75 to 671/mm3. Both the control group (n = 5) and the rifabutin group (n = 7) received 400 mg delavirdine mesylate every 8 h for 30 days; subjects in the rifabutin group took a 300 mg, once-daily dose of rifabutin on study days 16-30. Harvested plasma from serial blood samples collected after dosing on days 15, 16, and 30 was assayed for delavirdine and its N-desalkyl metabolite concentrations using a reversed-phase HPLC method. Blood samples obtained on days 16 and 30 were also assayed for rifabutin by HPLC. Delavirdine mesylate alone or in combination with rifabutin was well-tolerated. On day 30, statistically significant differences between groups were observed for all delavirdine pharmacokinetic parameters (P < 0.046). After coadministration of rifabutin and delavirdine mesylate for 2 weeks, oral clearance of delavirdine increased five-fold, resulting in lower steady-state plasma delavirdine concentrations. Rifabutin pharmacokinetic parameters were similar to those previously reported. Concomitant use of delavirdine and rifabutin at the recommended dose for each drug is discouraged. Maintaining therapeutic concentrations of delavirdine in patients on both medications may require dose modification. PMID- 9224962 TI - Developmental specificity of immunoglobulin heavy chain switch region recombination activities. AB - To understand the regulation of enzymes that carry out immunoglobulin heavy chain class switch recombination, we have assayed recombination of extrachromosomal substrates carrying switch region sequences in cell lines representing different stages of lymphoid cell development. Both pre-B and mature B cell lines supported switch substrate recombination, but B cell lines derived from later stages of cell development did not. Recombination did not occur in an erythroid or a macrophage line. Most recombination junctions in the substrates recovered from transfection of pre-B and B cells mapped to heterogeneous sites within the S mu and Sgamma regions, as do chromosomal switch junctions. Some recombination did occur in T cell lines, but most recombination junctions involved an upstream promoter and did not map preferentially to S regions. Culture of the pre-B cell lines PD31 and 70Z/3 with LPS increased recombination two-fold, to levels approaching those observed in LPS-cultured primary B cells. These results show that the full complement of factors necessary for switch recombination is present only in cells representing a limited spectrum of B cell development and that LPS, which can activate resting splenic B cells to carry out chromosomal recombination, can also stimulate recombination activity in immortalized pre-B cell lines. PMID- 9224963 TI - Purification and characterization of the immunoglobulin switch sequence-specific endonuclease (Endo-SR) from bovine spleen. AB - Mature B lymphocytes are able to specifically alter their Ig isotype expression in response to extracellular stimuli via a highly regulated, deletional recombination process called isotype switch recombination. Switch recombination breakpoints predominantly map to large (1-10 kb), G-rich and highly repetitive switch regions that are located directly upstream of immunoglobulin heavy-chain constant region genes. Switch region repeat structures vary considerably both within and between species, but all switch regions contain disproportionate numbers of two pentamer motifs, TGGGN and TGAGC, that are found at or directly adjacent to most analysed switch junctions. We have recently identified an endonuclease activity, Endo SR, that preferentially cleaves TGGGN and TGAGC switch motifs. We have purified the bovine endonuclease activity to homogeneity and have identified a protein with a molecular weight of approximately 32,000 that directly correlates with enzyme activity. As discussed in this report, we have found that murine and bovine Endo-SR are preferentially enriched in lymphoid tissue nuclear extracts and that both enzymes demonstrate highly similar physical and biochemical characteristics. However, each enzyme demonstrates related but distinctive specificities for consensus and degenerate TGGGN and TGAGC switch pentamer motifs. PMID- 9224964 TI - Evidence for phosphatidylinositol 3-kinase-dependent T cell antigen receptor (TCR) signal transduction. AB - Recent evidence implicates PI 3-kinase in TCR signal transduction. The fungal metabolite wortmannin is a specific inhibitor of PI 3-kinase both in vitro and in vivo when used at nanomolar concentrations. Therefore, we examined the effect of wortmannin on stimulation of primary T cells and T cell lines. Wortmannin had a dose-dependent inhibitory effect on TCR-dependent primary T cell proliferation with IC50 in the nanomolar range. Furthermore, activation of T cell lines independently of antigen presenting cells and, therefore of any CD28 co stimulatory signaling, was also sensitive to wortmannin. As expected, phorbol ester stimulation bypassed PI 3-kinase signal transduction. Importantly, the effect of wortmannin correlated with inhibition of activation of PI 3-kinase in stimulated T cells. The earliest step in T cell activation, tyrosine kinase activation, was not significantly affected by wortmannin. We conclude that a wortmannin-sensitive enzyme, probably PI 3-kinase, acting downstream of tyrosine kinases, but independently of the phorbol ester activated pathway, is necessary for stimulation of T cells via the TCR, and that this requirement is independent of any role of PI 3-kinase in co-stimulation via CD28 coreceptor. PI 3-kinase is most probably involved in generation of 3-phosphorylated lipid products, and is not merely an adaptor. PMID- 9224965 TI - Anti-ganglioside monoclonal antibody AA4 selectively inhibits IgE-induced signal transduction pathways in rat basophilic leukemia cells. AB - In rat basophilic leukemia 2H3 (RBL-2H3) cells, mAb AA4 binds to two derivatives of ganglioside GD1b that associate with the Src family kinase p53/56lyn and a serine kinase. Pre-incubation of cells with mAb AA4 blocks immunoglobulin E (IgE) mediated histamine release. In the present study we investigated the effect of incubation with mAb AA4 on signal transduction events. In addition to stimulation of the high affinity IgE receptor (Fc epsilonRI), cells were also activated by the calcium ionophore A23187 and the acetylcholine agonist carbachol in RBL-2H3 cells transfected with the G protein-coupled m3 muscarinic receptor. Incubation of cells with mAb AA4 in a dose-dependent manner inhibited the following Fc epsilonRI-induced signal transduction events: the increase of intracellular free calcium, phosphoinositol breakdown, tyrosine phosphorylation of proteins including the beta- of Fc epsilonRI and secretion. However, there was no inhibition of degranulation or of these biochemical events when cells were stimulated with calcium ionophore or activated via a G protein-coupled pathway. Our results demonstrate that mAb AA4 selectively blocks Fc epsilonRI-induced cell activation at a very early step upstream of receptor tyrosine phosphorylation. As mAb AA4 has previously been found to bind to gangliosides associated with Fc epsilonRI, inhibition of very early biochemical events may be due to interaction of mAb AA4 with the Fc epsilonRI induced signal transduction cascade at the receptor level. PMID- 9224966 TI - Variable region gene segment utilization in rhesus monkey hybridomas producing human red blood cell-specific antibodies: predominance of the VH4 family but not VH4-21 (V4-34). AB - Structural analyses of human immunoglobulin gene segments from monoclonal cell lines provide valuable information regarding the antibody repertoire. This information, in conjunction with a nearly complete knowledge of the human immunoglobulin germline repertoire, now allows further investigation into the underlying molecular basis responsible for some of the observed biases found in the expressed repertoire. One human heavy chain variable region gene segment, V4 34 (VH4-21), is one of the most prevalent gene segments in the expressed repertoire. The overwhelming presence of the V4-34 gene segment suggests that it may play an important role in immune responses. However, there is currently little information regarding its presence and potential importance in nonhuman primates. In order to determine if this gene segment is used by lower primates in a similar manner we determined the molecular structure of the variable region gene segments that are expressed by macaque monoclonal heterohybridomas that are specific for human red blood cell antigens. Eleven of the 12 hybridomas are derived from Rhesus monkeys (Macaca mulatta) and one is from a cynomologous monkey (Macaca fascicularis), all of which have been immunized with human red blood cells. The predominance of a VH4-like family and the specific absence of a VH4-21 equivalent led us to further characterize the macaque VH4 gene family at the germline level. Therefore, germline gene segments from the macaque equivalent to the human VH4 gene family are also described. PMID- 9224967 TI - Determination of the N- and C-terminal sequences required to bind human IgE of the major house dust mite allergen Der f 2 and epitope mapping for monoclonal antibodies. AB - B cell epitopes of the major house dust mite allergen Der f 2 from Dermatophagoides farinae were analysed using deletion mutants of Der f 2 expressed as fusion proteins in Escherichia coli. The reactivities of these partial Der f 2 molecules to human anti-mite IgE antibodies in atopic patients and to murine anti-Der f2 monoclonal antibodies (mAbs) were examined by immunoblotting. A C-terminal deletion mutant of Der f 2, 1-123, had almost the same reactivity to human IgE as the whole Der f 2 (1-129) and an N-terminal deletion mutant of Der f 2 (25-129) still had weak reactivity. On the other hand, in two deleted Der f 2 molecules, 1-120 and 30-129, reactivity was lost in spite of long overlapping sequences. These results suggest that the human IgE antibodies to Der f 2 in atopic patient sera recognize the conformational structures dependent on the tertiary structure of Der f 2, including disulfide bond formations, rather than the contiguous sequences of amino acids. The sequences 1-24, 25-29 and 121-123 were revealed as the minimum N- and C- terminal amino acid sequences required for IgE binding. Contrastingly, all three murine mAbs bound to the smaller deletion mutants, 1-90 and 67-129, suggesting that the cores of the epitopes for these mAbs exist in the 24 amino acid sequence of Der f 2, 67-90 overlapping the sequential human IgE epitope on Der p 2, the equivalent allergen from Dermatophagoides pteronyssinus. These findings are important for the understanding of the antigenic structure of Der f 2 and for the manipulation of the allergen for immunotherapy. PMID- 9224968 TI - Structural basis for the interaction of superantigen with the alternative superantigen-binding receptor p85. AB - Superantigens are microbial products which bind both to the TCR beta-chain and, with moderate affinity, to MHC class II molecules. Class II-bearing cells bind the superantigen and present the superantigen to T cells expressing certain TCR beta-chain variable region alleles. We have previously reported that the superantigen staphylococcal enterotoxin B (SEB) binds with moderate affinity to the protein p85 expressed on COS-1, an African Green Monkey kidney fibroblast like cell line. In the present report we carry out a structural analysis to examine the basis for the interaction of superantigen to p85. We show that SEC1, SEC2, and SEC3 also bind to p85 based on inhibition of the binding of radiolabeled SEB. On the other hand, SEA, SED, SEE and toxic shock syndrome toxin 1 do not exhibit detectable binding. In an effort to characterize the structural basis for the SEB binding to p85, we have generated both amino- and carboxy terminal truncations of SEB expressed as fusion proteins with the maltose-binding protein of Escherichia coli. Our results show that the full-length SEB fusion protein and a truncation missing the 81 amino-terminal amino acids both compete successfully with native SEB for binding. On the other hand, carboxy-terminal truncations in which 19 or 34 residues are deleted both fail to compete for binding. These results are consistent with results which show that monoclonal anti-SEB antibodies specific for carboxy-terminal determinants block SEB binding to p85, but an amino-terminal mAb fails to exhibit any alteration in binding. These results suggest that residues at or near the carboxy-terminus of SEB play a role in binding to p85. PMID- 9224969 TI - The ability of peptides to induce cytotoxic T cells in vitro does not strongly correlate with their affinity for the H-2Ld molecule: implications for vaccine design and immunotherapy. AB - The hypothesis that the ability of a peptide to bind to a class I molecule correlates with its immunogenicity is controversial. In this paper we have measured the affinity constants of nine synthetic peptides, which have been previously identified as binding to H-2L(d) molecules, and have determined their immunogenicity in an in vitro cytotoxic T lymphocyte (CTL) induction assay. We find that six peptides bind with high affinity (K(a) > 10(7)/M); of these, four are of viral origin but only two elicit potent CTLs, one is a self peptide which is not immunogenic, while the sixth is of bacterial origin and also does not generate effective CTLs. Two peptides bind with intermediate affinity (K(a) > 10(6)/M); one of these elicits a moderate CTL response, while the other, a tumor derived epitope, is highly immunogenic. Intriguingly, the peptide with lowest affinity (p2Ca) is exceedingly effective at eliciting CTLs. The efficacy of peptides with modest affinity for their restriction elements appears to correlate well with the CTL precursor frequency. We have also examined intrinsic parameters of some of the peptides such as solubility and stability. Taken together, our results underscore the relevance of factors other than affinity which affect immunogenicity and which may be critical in the design of peptide-based vaccines as well as tumor immunotherapy approaches. PMID- 9224970 TI - [A retrospective view of tumor immunology]. AB - The story of tumor immunology includes periods of hope followed by ones of disenchantment as far as clinical applications are concerned. In antiquity, cancer was considered "contrary to Nature", a concept which was confirmed by Ehrlich at the beginning of our century when the layed down the foundations of immunology. The latter was defined as the defence against all "non-self" intruders, including cancer, as opposed to the protection of "self". This concept was further accentuated by the theory immune surveillance proposed by Burnet in 1969 which implicated a destruction of nascent neoplastic cells by T lymphocytes. To increase host defence was the basis of tumor immunotherapy with BCG, levamisol and other adjuvants. The appearance of the nude mouse, athymic, and yet free of spontaneous tumors, led to a new paradigm, the network theory proposed by Jerne. This was based on immunological homeostasis implicating that both "self" and "non self" can be rejected and tolerated. Cancer gradually ceased to be considered as "contrary to Nature". As for the proposed viral etiology of cancer which was the basis of the National Cancer Act signed by Nixon in 1971, this led to various breakthroughs and Nobel Prizes (Table 1), to discoveries such as reverse transcriptase, cellular oncogenes, tumor suppressor genes, which gave a new explanation for neoplastic transformation. The latter can now be considered as the consequence of a cascade of molecular events which include oncogene expression, anti-oncogene deletion, etc... converting, step by step, for instance, a polyp into a colon cancer and its metastases. The availability of monoclonal antibodies capable of attacking tumor cells did not lead to the expected success because of the complexity of the immune system. Attempts at a better understanding of the latter have led to a subdivision of the T lymphocyte CD4 population into Th1 and Th2. Th1 favor rejection (tumoral, fetal or of transplants) through the elaboration of IL-2, IFN and TNF while Th2 led to tolerance or acceptation through the production of IL-4, IL-5 and IL-10: both functions neutralize each other establishing a "normal" equilibrium Th1 vs Th2. This could explain the state of "tumor dormancy" or tumors in situ which are apparently quite frequent. That any immunological stimulation would cause these dormant tumors to proliferate is the basis of the immunostimulation theory proposed by Prehn and supported by the clinical observations of Stewart. This new concept has led some authors to propose that instead of destroying the tumor cells an attempt be made to maintain them in a state of dormancy in congenial company with normal cells. PMID- 9224971 TI - Evidence for immune facilitation of breast cancer growth and for the immune promotion of oncogenesis in breast cancer. AB - Autoimmune diseases have been extensively studied in man and experimental animal models and salient points are reviewed, as a clear understanding of the immune mechanisms involved is essential if one is to understand the potential of immune interactions with established cancer cells or in the premalignant period of hyperplasia. Such reactions may be of benefit to the host, with down regulation of tumor growth, or unfavourable, with facilitation of oncogenesis and cancer growth. In particular, evidence is cited that supports a beneficial effect of the host response to non-small-cell lung cancer and the association of a poor prognosis in established breast cancer caused by a heightened immune response to the tumor. Histologic evidence supports these conclusions, as do studies of specific and nonspecific immune reactivity in breast cancer patients. The potential for cytokines to stimulate breast cancer growth, increase angiogenesis and decrease cell adhesion is reviewed, also recent evidence for autologous lymphocyte stimulation of breast cancer. Parallels between immune promotion of breast cancer in mice, caused by the mouse mammary tumor virus, and the development of breast cancer in women are also reviewed. If the mouse model has relevance for human breast cancer, one could predict that there would be a reduced incidence of breast cancer in a population of chronically immunosuppressed women following organ transplantation. Such is the case. This finding, plus the fact that all treatments that have shown efficacy in breast cancer have one thing in common, they are immunosuppressive, strongly support the role of immune facilitation of breast cancer growth and immune promotion of oncogenesis in breast cancer in a substantial number of growth. PMID- 9224972 TI - [Role of connective tissue in the tumor-host relationship]. AB - In the embryo, both differentiation and temporospatial organization are regulated by the mesoderm. Some of these functions are expressed by the connective tissue during wound or organ repair and regeneration. The normal development of the latter depends on the epithelium-mesenchyme interrelationship and the formation of an adequate amount of stroma and a certain type of collagen or proteoglycans. Our hypothesis proposes that cancer is a regenerative process which has failed as a consequence of alterations in the connective tissue. The object of this paper was to investigate whether the connective tissue and the amorphous fundamental substance (SFA) are capable of regulating the proliferation and death of normal and tumor cells and to duplicate the mechanisms involved. The results obtained in vivo, ex-vivo and in vitro experiments indicate that following: 1) SFA exerts a direct and selective cytotoxic effect on tumor cells; 2) SFA reduces the proliferative capacity of normal and tumor cells; 3) both the cytotoxic and antiproliferative effects of SFA are independent of cellular and humoral immune responses but are dependent on the chemical integrity of its component since its denaturalization reduces its antitumoral activity; 4) the tumor cells modulate the regulatory effects of SFA through endocellular enzymes liberated by cell death induced by the cytotoxic action of SFA itself. These results suggest that in the absence of the inhibitory effect of SFA, the tumor cells which remain viable con now proliferate actively due to enzyme stimulation. In conclusion, the regulatory function of the connective tissue on the proliferation and viability of tumor cells would depend on the molecular constitution of SFA. PMID- 9224973 TI - [Immunoregulation of murine mammary tumor growth]. AB - The function of the immune system during tumor growth is very controversial. Although there were great hopes that a proper stimulation of immunity would erradicate tumor cell proliferation this objective has still not been attained. We review and analyze some of the immunologic studies performed in our laboratory using spontaneous murine mammary adenocarcinomas. In early stages of tumor growth our studies showed a specific immune response in vivo and in vitro by delayed type hypersensitivity tests, and syngeneic lymphocyte induced angiogenesis assay. These activated lymphocyte responses were not involved in mechanisms effective for tumor rejection. The lymphocyte activation vanished with tumor growth. Soluble factors shed from splenocytes of tumor bearing hosts and factors secreted by tumor cells enhanced tumor and metastatic growth. Other cell populations, such as mastocytes and neutrophils were also altered during tumor growth. We conclude that soluble factors released by tumor cells induce the production of suppressor cells or factors that stimulate tumor and metastatic growth. PMID- 9224974 TI - [Immunobiological characterization of murine LB leukemia and the LBC cell line]. AB - LB leukemia is a nonimmunogenic T cell tumor which spontaneously arose in a BALB/c mouse; efforts to induce immunological rejection of the leukemic cells have always failed. The leukemic cells grow rapidly and progressively in the syngeneic host invading spleen, lymph nodes and liver. A cell line (LBC) was developed from the original tumor. Both the original tumor and the cell line have been characterized as expressing the Thy 1+, CD3-, CD25+, MHC class I+, class II , CD4- (original tumor), CD4+ (cell line), CD8+, gp70-, J11d.2+ phenotypes. Immunization of syngeneic mice with irradiated LBC cells induced cytotoxic T lymphocytes as well as anti-LBC antibodies which reacted with components of 14, 16 and 27 kDa present on LB tumor cells, LBC cell line and normal thymocytes but not on normal lymph node cells. Immunization of syngeneic mice with LBC cells partially protected them against subsequent challenge with the original tumor cells. The effect of sera from tumor-bearing mice and the super-natants from short term cultures were studied on cell proliferation. An inhibitory activity was demonstrated in these fluids, which was abrogated by addition of exogenous IL 2. ELISA showed the presence of soluble IL-2R alpha chain both in the conditioned medium as well as in the serum, which was demonstrated to be responsible for the inhibitory activity. The soluble IL-2R was produced by LB leukemic cells and exerted the inhibitory activity blocking cell proliferation and modulating immune response by binding to free IL-2. Using reverse-transcription PCR, mRNA for IL-2 was found to be present in tumor cells. Our findings indicate that LB cell proliferation is mediated by an autocrine pathway involving endogenous IL-2 generation, despite the fact that these cells are not dependent on exogenous IL-2 to grow in culture. The relationship between tumorigenicity and expression of MHC class II was also investigated. In vitro treatment with IFN-gamma failed to induce the expression of class II antigens in LBC cell line. Therefore these cells were tri-transfected by a liposome-mediated protocol with 1-A alpha d, I-A beta d genes and pSV2neo. Cells were selected to grow in medium containing Genetecin (G418) and surviving transfectants were cloned. Three I-A+ clones were obtained (LBCT) and were used to induce a specific CTL response against tumor cells. Syngeneic mice inoculated with 10(3) LBCT cells failed to develop a tumor while the DT50 of mice injected with 10(6) LBCT cells was three times the value for mice injected with LBC cells (I-A-). It is suggested that neoexpression of MHC class II molecules enhances anti-tumor response by transforming tumor cells into professional antigen-presenting cells, which may be used to improve tumor specific immunity in the autologous host. PMID- 9224975 TI - [Concomitant antitumor resistance]. AB - Concomitant resistance of tumor-bearing mice against a second tumor challenge was evaluated in euthymic and athymic mice using 17 tumors with different degrees of immunogenicity. Two temporarily separated peaks of concomitant resistance were detected during tumor development: the first peak was only observed associated with small immunogenic tumors (< 500 m3., it was tumor-specific and mediated by T cell-dependent immunological mechanisms. The second peak was exhibited by large tumors (> 2000 mm3) independently of their immunogenicity; it was non-tumor specific, thymus-independent and correlated with a serum-activity (neither antibodies nor complement) which inhibited the in vitro proliferation of tumor cells. Out of 17 tumors studied, 15 tumors exhibited a moderate or strong concomitant resistance. The remaining two, which exhibited a weak or undetectable concomitant resistance and correlatively, a low or absent serum-inhibitory activity were the only tumors which included lung metastases. This fact suggested a correlation between concomitant resistance, absence of metastases and the existence of an inhibitory factor(s) in the serum. This inhibitory factor was partially characterized: it was resistant to boiling (5-10' at 100 degrees C) and to variations of pH; its molecular weight was estimated between 850 and 1200 D; it was recovered in only one fraction from HPLC (high power liquid chromatography) columns presenting maximum absorption at 215 and 266 nm; amino acid analysis and magnetic nuclear resonance studies suggested the presence of a molecule of thyrosine and one or two molecules of carbohydrates in its structure. PMID- 9224976 TI - Immunostimulation of cancer versus immunosurveillance. AB - A moderate antitumor immune reaction is optimal for tumor growth and most (perhaps all) untransplanted tumors, rather than being inhibited, are probably dependent, at least early in their progression, upon the immune reaction. Some tumors, as for example most human skin tumors, have a higher incidence in immunodepressed patients than they do in the general population. This could mean that the normal immune reaction usually inhibits the growth of these tumors. More probably, the increased incidence in immuno-depressed heart and kidney transplant patients is caused by a lowering of a tumor-stimulatory immune reaction to a level that is even more stimulatory. Other tumors, such as human mammary tumors, have, in contrast to the skin tumors, a lower than expected incidence in immunodepressed patients. Mammary tumors are postulated to possess, on average, a low immunogenicity that arouses an immune reaction that is usually equal to or less than that required for optimal tumor growth; a further lowering of the reaction, as occurs in heart or kidney transplant patients, thus results in a lowered tumor incidence. In such patients, an immunostimulating adjuvant or vaccine might, we predict, sometimes accelerate rather than inhibit tumor growth. PMID- 9224977 TI - Immune mechanisms and tumor dormancy. AB - The natural occurrence of tumor dormancy in man is reviewed based on observations made by a distinguished surgeon and two pathologists after a lifetime of practice. They concluded that dormancy is the result of immune mechanisms preventing the growth of microscopic metastases already present following curative surgery. Six cases of non-small-cell lung cancer are described in patients randomized to receive specific active immunotherapy in controlled clinical trials. Three patients had nonregional metastases at 11, 12 and 14 years. Two had regional recurrence after 9 years and in one patient a small hilar node metastasis was found at necropsy after 7.6 years. In each case an immunodepressive event or drug treatment preceded resurgent growth. Animal models of tumor dormancy are reviewed and the evidence is clear that dormancy may be induced by manipulating immune mechanisms, resulting in cells remaining in mitotic arrest, or tumor cell proliferation is balanced by an equivalent rate of cell death. Based on these observations future clinical strategies should de developed to induce the dormant state in micrometastases in the adjuvant setting. PMID- 9224978 TI - [Spontaneous regression of cancer]. PMID- 9224979 TI - [Melanoma: model and opportunities for immunologic intervention]. PMID- 9224980 TI - [Immunotherapy in melanoma and renal tumors]. PMID- 9224981 TI - [Treatment of multiple myeloma with interferon]. PMID- 9224983 TI - [Perinatal and neonatal mortality]. PMID- 9224982 TI - [Digitalis and its current importance in the treatment of heart failure]. PMID- 9224984 TI - [Severity of acquired cystic kidney disease determines improvement in chronic kidney failure anemia]. AB - OBJECTIVE: To identify a correlation between dialysis treatment duration and severity of acquired cystic disease of the end-stage kidney measured by cyst sizes; and assess its effect on spontaneous improvement of anemia. MATERIALS AND METHODS: Ten patients, 6 males and 4 females were selected, who have been on hemodialytic treatment for more than 5 years. There was no patient selected with autosomal dominant polycystic kidney disease. The renal evaluation has been made through ultrasonographic studies. Acquired cystic disease of the end-stage kidney was characterized by finding four or more cysts in both kidneys. The largest cysts were measured for correlation effect. The diagnosis of anemia was established by hematocrit and hemoglobin serum values. It has also been analyzed serum urea, creatinine, albumin, iron, total iron-binding capacity and the per cent saturation of serum transferrin were tested. RESULTS: The patients were properly dialysed (serum urea and creatinine = 98.7 +/- 35 mg/dL e 9.7 +/- 2.7 mg/dL, respectively). There were in good nutritional shape (serum albumin = 4.5 +/- 0.5 g/dL) and had normal serum iron level (serum iron = 80 +/- 34 mg/dL). The prevalence of acquired renal cystic disease was 80%. No finding of malignancy has been detected in these cysts. There was a significant correlation between time on dialysis treatment and hematocrit values (R = 0.70; p < 0.05). Cyst sizes had a direct and significant correlation with hematocrit levels (R = 0.74; p < 0.05). CONCLUSION: These results show that spontaneous improvement on anemia seen in patients on chronic dialysis has a significant correlation with the severity of acquired cystic disease of the end-stage kidney. Our data suggest a functional role of acquired kidney cysts on endogenous erythropoietin production. PMID- 9224986 TI - [Bacterial aerosol generated by mechanical ventilators: comparative study]. AB - Mechanical ventilators generate aerosol which may be bacterially colonized. PURPOSE: To determine the environmental contamination generated by ventilators with two different humidification techniques. METHODS: The study was done comparing the generation of bacterial colonized aerosol by the expiratory valve of mechanical respirators with conventional water nebulization or with hygroscopic condensator as the humidifier source during 15 minutes of observation. RESULTS: The aerosol got positive cultures in 32.2% of the conventional system and in 5% of the condensator system (p = 0.0340). CONCLUSION: We concluded that the humidification by the hygroscopic condensator may be an efficient way to reduce environmental bacteria contamination. PMID- 9224985 TI - [Clinical significance, epidemiology and microbiology of coagulase-negative staphylococcal nosocomial bacteremia at a teaching hospital]. AB - Coagulase-negative staphylococci (CNS) are an important cause of nosocomial bacteremia and they are frequently considered as contaminants of blood-cultures. From October 1990 to September 1992, 300 positive blood-cultures for CNS at the Hospital Sao Paulo were studied and 141 CNS bacteremias were characterized as nosocomial bacteremias. Clinical and microbiological criteria were defined to differentiate between true CNS bacteremia and contaminated cultures. Only 20.6% of the CNS nosocomial bacteremia were considered as true bacteremia. Most of the CNS true nosocomial bacteremia were detected among newborns admitted to the neonatal intensive care unit; the presence of intravascular catheter and parenteral nutrition were significant findings. We did not detect significant difference between true nosocomial bacteremia and contaminated cultures regarding to resistance to oxacillin and SLIME production. The clinical criteria and the positivity of the blood-cultures up to 48 hours after incubation, utilized in our definitions, were useful parameters to characterize the CNS true nosocomial bacteremia. PMID- 9224987 TI - [Endocervical tubal metaplasia: morphological concepts and practical importance]. AB - OBJECTIVE: Among uterine cervix tumor-like lesions, tubal metaplasia (TM) has been confused with endocervical in situ adenocarcinoma. TM is a benign lesion composed of three cellular types: ciliary, secretory and intercalary (or peg cell). Thus, the main purpose of this work is to localize and characterize tubal metaplasia and its relation to other morphological lesions in the cervix. METHODS: Eighteen cervical specimens from 8 cones and 10 hysterectomies with TM were reviewed in order to observe its relative frequency in different segments such as: superior, inferior, surface epithelium and glands. All cases were associated to other neoplastic and non-neoplastic diagnosis. RESULTS: TM was observed in cases with an age span from 24 to 72 years old, with a mean age of 44 years. In most of the cases (83%), TM was found in the superior region of the cervix, but in 61% TM was also found in the inferior region, either on the surface epithelium or in the glands. In 60% of the cases TM was associated with invasive or intra-epithelial neoplasia. CONCLUSIONS: In spite of being observed in the higher parts of the endocervix, TM was also detected in the lower segment, where the differential diagnosis with in situ adenocarcinoma is important. Thus, although more frequently associated with neoplasia in this study, it is not possible to determine the real incidence of TM in the cervix. However, morphological characterization of the lesion is very important for diagnostic purposes. PMID- 9224988 TI - [Increase in the search for treatment by crack users in 2 outpatient clinics at the city of Sao Paulo from 1990 to 1993]. AB - An increase in crack use epidemiological research and police data. Currently, in Brazil, no data are available linking the route of administration and attendance to treatment for cocaine dependence. OBJECTIVE: The purpose of this paper was to analyze the changes in cocaine routes of administration in a cocaine dependent population treatment in two outpatient public services (PROAD and UDED). METHOD: Standardized interview data, collected at admission to treatment were compared from 1990 to 1993. The prevalence rates of smoked ("crack"), injected and snorted cocaine were compared. RESULTS: The percentage of patients who reported "crack" cocaine use increased from 17% in 1990 to 64% in 1993 (p < 0.01) The prevalence of snorted cocaine remained stable in the period of time analyzed, being the most frequent route reported. The intravenous route tended to decrease from 40% in 1990 to 28% in 1993. CONCLUSION: The implications of the increase of "crack" cocaine users who sought treatment are discussed. These data are important in planning prevention and treatment strategies, mainly in AIDS prevention. PMID- 9224989 TI - [Immunologic fetal and neonatal immaturity: influence on the clinical course of HIV-1 infection in children]. AB - Children born to HIV-1 infected mothers present a more severe clinical evolution than adults or children infected by other routes. The physiologic immaturity of the fetal and neonatal immune systems at the time of the infection probably plays an essential role in the progression of HIV-1 infection in these children. This paper describes the development of the normal human immune system and its correlation with the immunopathogenicity of vertical acquired immunodeficiency syndrome (AIDS). PMID- 9224990 TI - [Perinatal and neonatal mortality at the Clinicas de Porto Alegre Hospital, Brazil]. AB - OBJECTIVE: Epidemiological analysis of neonatal and perinatal mortality of 20,280 newborns alive with 500g or more and 374 stillbirths occurred at the Hospital de Clinicas de Porto Alegre from 1984 to 1990. PURPOSE: To compare two periods: A (1984-1987) with B (1988-1990), establishing a relationship between the changes occurred in the causes and the rate of mortality. METHODS: The retrospective study was done with the records of promptuaries of obstetrical and neonatal centers, and review of flow-sheets of the deaths and autopsies. RESULTS: Between 1984 to 1990, 20,280 newborns alive with 500g or more, 374 stillbirths at perinatal unit of Hospital de Clinicas de Porto Alegre were born. 258 deaths occurred, the neonatal mortality rate was 12.7 per thousand. The stillbirth rate was 18.4 per thousand. The perinatal mortality rate was 28.4 per thousand. The incidence of low birth weight (< 2,500g) was 11.2% and very low birth weight (< 1,500 g) was 1.8%, the former group had an increase incidence between 1984-1988 (A) from 1.5% to 2.2% (B). The causes of deaths were distributed as follow: a) intrauterine infections (22.4%); b) hyaline membrane disease (20.1%); c) congenital malformation (18.2%); d) asphyxia (15.5%); e) postnatal infections (9.7%). The causes of stillbirth were: a) perinatal asphyxia (38.7%); b) intrauterine infections (9%); toxemia (8.2%); d) malformation (7.4%). The period B showed changes with an increase of postnatal infections odds ration (OR) 7 (1.9 30.6) and congenital malformations OR 1.6 (0.8-3.2). It did not occurred a decrease in mortality rate for prematures below 1,500g OR 90 (61-118) in A to 54 (37-68) in B. CONCLUSIONS: The advantages in technology and human capacity were not sufficient to reduce significantly the rate of neonatal mortality. PMID- 9224991 TI - [Study of bone density in systemic scleroderma]. AB - BACKGROUND: Osteopenia in systemic sclerosis (scleroderma) patients was reported in X-ray studies of hands and by proximal and distal forearm bone mass measurement. It has been suggested that bone loss in these patients might be due to chronic ischemia, immobilization and early menopause. Nevertheless, it is not established if these patients present generalized osteopenia. To shed light into this point we studied bone mineral density in the spine, proximal femur and total body in patients with systemic sclerosis. PATIENTS AND METHOD: Twenty-five Caucasian women were evaluated. Mean age of patients was 48 +/- 12 years and mean time of disease was 7 +/- 7 years; 13 were postmenopausal (8 +/- 8 years). Bone mass was measured in the spine, proximal femur and total body by using a dual photon absorptiometry with X rays source (Lunar-Model DPX). RESULTS: Bone mass in different sites was not statistically different from the age-matched control healthy women. Mean bone mass of patients with limited form was not different from patients with diffuse form of systemic sclerosis. Patients with calcinosis had lower bone mass at proximal femur than those without this alteration. CONCLUSIONS: Patients with systemic sclerosis do not present bone loss and this disease in not a risk factor for generalized osteoporosis. PMID- 9224992 TI - [In vitro activity of cefetamet compared with other antimicrobial agents against bacteria isolated from respiratory tract infections]. AB - Cefetamet pivoxil is a new beta lactamase orally stable administered cephalosporin. Antimicrobial resistance among respiratory pathogens has become an important problem for both the physician and the microbiologist and the patterns of resistance vary greatly depending on geographic location, often requiring in vitro susceptibility testing of isolates. PURPOSE: The in vitro activity of cefetamet, the microbiologically active metabolite of the prodrug cefetamet pivoxil, was compared with other 11 drugs against 376 bacterial strains recently isolated from patients with respiratory tract infections. METHODS: The comparative activity in vitro of cefetamet and other 11 antimicrobial agents was measured against 376 bacterial strains isolated from patients with respiratory tract infections, during a six month period. Through the determination of minimum inhibitory concentration by the microdilution technique, patterns of antimicrobial resistance were reported. RESULTS: Cefetamet showed high in vitro activity against all the bacterial tested, possessing a spectrum of activity similar to that other recently developed oral cephalosporins. The good activity of cefetamet against beta-lactamase producing isolates, like Moraxella catarrhalis, can be due to its beta-lactamase stability. At a concentration of 1.0 microgram/mL, cefetamet inhibited 97% of all the tested bacteria. CONCLUSION: The MIC90 of the cumulative susceptibility results of the 12 antimicrobics tested in the 376 strains studied, confirm the excellent activity of cefetamet against the common respiratory tract pathogens. PMID- 9224993 TI - [Cost of liver transplantation at the Clinical Hospital of the University of Parana, Brazil]. AB - PURPOSE: To determine the cost of liver transplantation at the Clinical Hospital of the Federal University of Parana. METHODS: The data of 24 patients subjected to 25 liver transplantations were evaluated from the day of hospital admission until the day of discharge to determine the length of hospitalization, quantity of material and medications used, and exams and procedures performed. Professional fees were not included in the study. RESULTS: The age of the patients varied from 6 to 56 years. Six patients were younger than 14 years of age. Five patients died during hospitalization. Retransplantation was performed in only one patient. The average cost for liver procurement was US$ 2,783.19. The total cost of the liver transplantation varied, depending on the occurrence of complications, length of hospitalization and the amount of blood products transfused. The total cost varied from US$ 6,359.84 to US$ 75,434.18, with an average of US$ 21,505.53. The most expensive item of the liver transplantation was blood products transfused, followed by medications, and intensive care and room charges. CONCLUSIONS: The cost of liver transplantation varies among the patients and may be performed in Brazil at a cost less than that reported in the United States and Europe. PMID- 9224994 TI - [General characteristics of the spermatozoa in oligozoospermic men with and without clinical varicocele]. AB - Patients with oligozoospermia show a reduction in the semen quality, independent of the etiology of the disturbance. PURPOSE: To investigate the role of the varicocele in the decrease of the semen quality in oligozoospermic men. METHODS: Ten patients with left clinical varicocele (termed PCV) and 21 patients without this entity (termed PSV) attended in a private laboratory from Petropolis, RJ, were investigated. Sperm count, vitality, motility, and morphology of spermatozoa were examined and the results were compared between them and a control group consisting of 15 patients without clinical varicocele and with normal spermiogram. RESULTS: PCV and PSV had showed significant decrease in the vitality (43.9% and 34.9% versus 73.0% in the control group), grade (a) (5.3% and 2.4% versus 32.4% and grade (d) (76.7% and 83.8% versus 44.9%) of sperm progression and in the percentage of oval sperms (25.5% and 22.9% versus 61.2%), amorphous head (25.4% and 23.8% versus 12.5%) and other anomalies (23.8% and 30.5% versus 13.0%). PCV had also showed significant difference in the percentage of tapered sperm (10.9% versus 1.3%), whereas PSV had showed significant difference in the grade (b) of sperm motility (11.0% versus 22.0%), both in regard to the control group. Between PCV and PSV had not been found significant differences. CONCLUSION: Varicocele reduces the semen quality in oligozoospermic men, but this reduction also occurs in oligozoospermia of any etiology. PMID- 9224995 TI - [Free radicals: concepts, associated diseases, defense system and oxidative stress]. PMID- 9224996 TI - [Panoramic nailfold capillary microscopy and its application in rheumatic diseases]. PMID- 9224997 TI - [Antivenom serum doses in the treatment of poisoning by a venomous snake of the genus Bothrops]. PMID- 9224998 TI - [Molecular biology of Alzheimer's disease: a light at the end of the tunnel?]. PMID- 9224999 TI - [Polyneuropathy deficiency among Xavante indians]. AB - The authors present two cases of polyneuropathy deficiency among Xavante indians where the sole food was rice in case 1 and almost so in case 2. The rice consumed by these indians was processed or hulled. Intoxication by cyanide from maniot or other vegetable was excluded. CASE REPORT: Two indians aged 18 and 25 years with a progressive history of weaness, decrease in muscular force and thinning were observed in their villages. On removal to the Hospital Sao Paulo, atrophy of the distal musculature of the upper and lower limbs, motor deficit distally with zero degree in the flexor musculature, abolished deep reflexes, plantar cutaneous reflex without response bilaterally, decreased tactile, painful and pallesthetic sensitivity distally in the lower limbs were noted on neurological examination of case 1. On neurological examination of case 2 proximal hyporeflexia in the upper limbs, areflexia in the distal portions of the upper and lower limbs, tactile and painful hypoesthesia in the feet, right hypoacousis were noted. Electromyography showed abnormalities compatible with symmetric sensorimotor polyneuropathy with an axonal demyelination pattern in case 1 and predominantly demyelinizing in case 2. Cerebrospinal fluid tests were normal. DISCUSSION: Polyneuropathy was characterized by the clinical history and by neurological, electromyographic and cerebrospinal fluid tests. The diagnosis of polyneuropathy deficiency was established by the clinical history and by electromyography suggesting peripheral polyneuropathy of nutritional origin. This neuropathy deficiency does not fit myeloneuropathies such as ataxic tropical neuropathy, spastic paraparesis and Cuba neuropathy. CONCLUSION: The Xavante polyneuropathy deficiency is caused by thiamine (vitamin B1) deficiency, that is dry beriberi, owing to consumption of industrially processed rice as sole or almost sole food. The Xavante polyneuropathy is different from the neuropathy observed among Kreen-Akrore indians and from that of adolescent indians in the Xingu Park. PMID- 9225000 TI - Treatment of murine gliomas by adoptive transfer of ex vivo activated tumor draining lymph node cells. AB - The adoptive transfer of tumor-reactive T lymphocytes has recently been demonstrated to be an effective means for mediating the regression of experimental intracranial fibrosarcomas. In this study, mice bearing syngeneic intracranial GL261 gliomas were cured by the combination of sublethal whole body irradiation followed by the intravenous transfer of tumor-draining lymph node (LN) T cells activated with anti-CD3 or staphylococcal enterotoxin C2 (SEC2). To further identify the functional effector T cel population in the adoptive immunotherapy, LN T cells were separated into two subsets, based on the level of expression of the cell adhesion molecule CD62L (L-selectin). As few as 5 x 10(5) CD62Llow cells could cure the majority of animals, whereas 2 x 10(6) CD62Lhigh cells were completely ineffective. Moreover, T cells isolated from advanced intracranial tumors were identified to be predominantly CD62Llow. In contrast, spleens contained a mixture of CD62L low and high cells similar to the transferred cell population. T cells in the glioma site were more actively proliferating than those isolated from the spleen. Mice cured of GL261 tumors demonstrated long-term immunologic memory by rejecting intracranial challenges of the original tumor but not an immunologically distinct tumor. Furthermore, despite infiltration of transferred cells into the intracranial tumors, cured mice did not exhibit any apparent neurologic abnormalities during treatment, prolonged follow-up, or after intracranial tumor rechallenge. This study demonstrates the effective treatment of an intracranial murine glioma by the systemic adoptive transfer of activated tumor-draining LN T cells and selective tumor infiltration by the therapeutically active CD62Llow T cells. PMID- 9225001 TI - Glucocorticoid regulation of natural cytotoxicity: effects of cortisol on the phenotype and function of a cloned human natural killer cell line. AB - The ability of glucocorticoids to suppress cellular immune functions, including the cytotoxic activity of natural killer cells, is well known. However, the molecular mechanism(s) of glucocorticoid-mediated suppression of cellular cytotoxicity mediated by natural killer cells is not understood. We have investigated the effects of cortisol on protein expression and cytotoxic function of natural killer cells using NK3.3, a well-characterized, cloned human natural killer cell line. Cortisol, at concentrations up to 2 microM, does not significantly alter the viability or proliferative capacity of NK3.3 cells. However, micromolar concentrations of cortisol induce the expression of a small set of proteins which are not synthesized by NK3.3 cells in the absence of cortisol, and repress the synthesis of another set of proteins including several phenotypic determinants and cytokines. In the presence of added cortisol, the synthesis of perforin mRNA was partially repressed. However, the most striking effect of cortisol on this NK clone was its repression of granzyme A synthesis. In conjunction with the downregulation of adhesion proteins, NK3.3 cells cultured in the presence of cortisol exhibit a reduced capacity to form conjugates with K562 target cells. Whereas cortisol treatment of NK3.3 cells causes an approximately 50% decrease in their ability to form conjugates with K.562 target cells, the cytotoxic function of these cells is completely abolished under the same conditions. This first report of hormonal regulation of granzyme expression and the strong correlation between granzyme A repression and cytotoxic function suggests that cortisol may regulate NK function by repression of granzyme A synthesis. In addition to demonstrating the significant influence of cortisol on natural killer cell function, these studies provide a model system for elucidation of molecular mechanism(s) whereby glucocorticoids repress cellular immune function, especially with respect to natural killer cells. PMID- 9225002 TI - Early steps in T cell development are affected by aging. AB - Involution of the thymus accompanies aging, a process in which the organ diminishes in size and cellularity and becomes disorganized. The rate of T cell emigration from the thymus is markedly reduced with age, and phenotypic analyses have identified alterations in the relative proportions of the major thymocyte subpopulations. The present studies made use of the capacity of the thymus to regenerate following irradiation from an intrathymic radio-resistant precursor population. By analysis of the differentiation of this "wave" of thymocytes, it was determined that aging most severely affects the earliest developmental transitions. While the overall rate of differentiation does not appear to be affected in older mice, fewer thymic progenitors initiate differentiation. The reduced expansion of late pre-T cells in the middle-aged is due to the smaller pool size of these cells. PMID- 9225003 TI - Short-term dexamethasone treatment modulates the expression of the murine TCR zeta gene locus. AB - Glucocorticoids (GCH) are highly effective agents in controlling inflammation and immune response. We studied the effect of the synthetic GCH dexamethasone (DEX) on the expression of TCR zeta gene splicings that code for some chains belonging to the T-cell receptor (TCR)/CD3 complex. In the DEX-treated hybridoma T-cell line 3DO, TCR zeta gene splicings increase within the first 24 hr (about fourfold increase), as demonstrated by reverse transcriptase-polymerase chain reaction and RNase protection assay. This increase is due to the stimulation of TCR zeta gene locus transcription, as demonstrated by the "run-on" assay. A similar upregulation was observed in murine thymocytes following in vivo DEX treatment. As a consequence of TCR zeta gene locus modulation, the expression of the spliced mRNAs coding for TCR zeta and TCR eta subunits is increased, whereas their relative ratio is only slightly changed. Indeed, the amount of TCR zeta protein in 24-hr DEX-treated cells is fivefold more than that in the untreated cells. A similar effect was seen in 3DO cells treated with hydrocortisone but not in those treated with testosterone. TCR zeta protein increase was confined to the cytoplasm and therefore TCR/CD3 complex expression did not increase. This newly described effect of DEX may constitute an additional molecular mechanism that contributes to its immunomodulating activity. PMID- 9225004 TI - Ligation of CD53/OX44, a tetraspan antigen, induces homotypic adhesion mediated by specific cell-cell interactions. AB - The CD53 antigen is a member of the tetraspan family of proteins with unknown function. Stimulation of rat IR938F B-cell lymphoma cells with monoclonal antibody MRC OX44 (anti-rat CD53) triggered a homotypic adhesion reaction which reached a maximum effect at 24 hr. This effect occurred at 37 degrees C but not at 4 degrees C. Adhesion was prevented by removal of divalent cations, Ca2+ and Mg2+, with EGTA and EDTA as chelating agents. The adhesion induced by MRC OX44 was inhibited by cycloheximide and actinomycin D, suggesting that de novo protein synthesis was required for this effect. The addition of mAb WT1 against rat LFA-1 (CD11a) antigen had no effect on adhesion, suggesting that the cell-cell interaction is not mediated by the expression of LFA-1 antigen. The intracellular signals required to induce adhesion were inhibited by two tyrosine kinase inhibitors, genistein and piceatannol. Wortmannin, a selective inhibitor of phosphoinositide 3-kinase activity, completely blocked adhesion. Two protein kinase C inhibitors, H7 and bisindolylmaleimide, inhibited the adhesion, suggesting that part of the signal is mediated by PKC. Electron microscopy of aggregated cells showed that the interaction is localized to short membrane regions, where contact areas of higher density in opposing zones from both cells were detected. We postulate that there is a common adhesion mechanism that is modulated by several tetraspan family members and associated proteins. This adhesion structure might represent a novel form of cell communication among lymphoid cells. PMID- 9225005 TI - Posttranslational regulation of Lck and a p36-38 protein by activators of protein kinase C: differential effects of the tumor promoter, PMA, and the non-tumor promoter, bryostatin. AB - T cell activation via the antigen receptor or by PKC-activating drugs results in phosphorylation of Lck and alteration of its electrophoretic mobility. Although tyrosine phosphorylation appears to regulate Lck enzymatic activity, the significance of phosphorylation of serine residues and its relevance to the cell proliferation process are yet unclear. We found that the PKC activator, bryostatin, like PMA, induced the conversion of p56lck to a slower migrating form with an apparent molecular mass of 60 kDa. The effect of PMA lasted over 48 hr but that of bryostatin was transient and correlated in time kinetics with that of the bryostatin-induced degradation of PKC. The effects of bryostatin were dominant over those of PMA. In addition, PKC was found to affect both serine and tyrosine phosphorylation of Lck but had no significant effect on the in vitro catalytic activity of Lck. To test whether serine phosphorylation of Lck may affect its ability to bind tyrosine phosphoproteins, we compared Lck immunoprecipitates from PMA- and bryostatin-treated T cells. We found that a 36- to 38-kDa tyrosine phosphoprotein co-immunoprecipitated with Lck from cells that were treated for 24 hr with PMA, but not bryostatin. A p36-38 from PMA- but not bryostatin-treated cells also interacted with an Lck-SH2 fusion protein, suggesting differential regulation of p36-38 by PMA and bryostatin. Furthermore, in vitro phosphorylation of p36-38 occurred in lysates of cells that were treated for 24 hr with PMA, but not in lysates of bryostatin-treated cells. The results show that tyrosine phosphorylation and the association of p36-38 with Lck are differentially affected by bryostatin and PMA and suggest that PKC regulates the interaction of potential signaling molecules with Lck, thereby regulating biochemical events that are relevant to T cell mitogenesis and/or transformation. PMID- 9225006 TI - Potentiation of antitumor CTL response by GM-CSF involves a B7-dependent mechanism. AB - We have previously demonstrated the importance of endogenous GM-CSF production for the B7-2-dependent potentiating effect of exogenous TNF for CTL generation by stimulation cultures of splenic cells from mice bearing a large MOPC-315 tumor. Here we show that addition of GM-CSF to stimulation cultures of such tumor-bearer splenic cells also leads to the generation of enhanced anti-MOPC-315 CTL activity via a B7-dependent mechanism. However, while the potentiating effect of TNF was previously shown to be IL-2-independent, the potentiating effect of GM-CSF is shown here to be completely IL-2-dependent. Still, the potentiating activity of exogenous GM-CSF for the in vitro generation of CTL activity is shown to depend completely on endogenous TNF production. Finally, TNF and GM-CSF may cooperate in enhancing the in vivo generation of CTL activity in MOPC-315 tumor bearers because low-dose melphalan (L-phenylalanine mustard) therapy, which was previously shown to lead to the rapid up-regulation of TNF production at the tumor site and the subsequent TNF-dependent in vivo acquisition of potent CTL activity, is shown here to lead to the rapid up-regulation of GM-CSF production at the tumor site. PMID- 9225007 TI - Recognition of gp43 tumor-associated antigen peptide by both HLA-A2 restricted CTL lines and antibodies from melanoma patients. AB - Previously, we detected a 43-kDa tumor-associated antigen (TAA) using the human monoclonal antibody L92, which recognizes the tetramer peptide KYQI. In the present study, cell lines of cytotoxic T lymphocytes (CTL) specific to the gp43 peptide (DLTMKYQIF) were established from peripheral blood lymphocytes (PBL) of melanoma patients. Patients' PBL (n = 326) of different HLA Class I types were assessed for gp43 CTL activity. CTL specific to gp43 peptide were generated only from HLA-A2 melanoma patients and not normal donors. gp43 CTL recognized gp43 peptide-pulsed autologous BLC and T2 HLA-A2 target cell lines. Furthermore, CTL lines were shown to kill both HLA-A2 autologous and HLA-A2 allogeneic melanoma cell lines, indicating that gp43 peptide can be processed endogenously and presented by melanoma cells as a common TAA. The gp43 CTL lines did not kill normal cells. Specific amino acids of the peptide were shown to be important determinants in stimulation and recognition of CTL. gp43 peptide, recognized by both antibodies and T cells of melanoma patients, is a novel TAA peptide that may play an important role in anti-tumor immunity in human. PMID- 9225008 TI - Spontaneous hair follicle cycling may influence the development of murine contact photosensitivity by modulating keratinocyte cytokine production. AB - The development of murine contact hypersensitivity is influenced by hair follicle cycling. Here, we have examined hair cycle-associated fluctuations of murine contact photosensitivity (CPS) to tetrachlorosalicylanilide (TCSA) and its immunologic mechanism(s). When the CPS outcome was monitored in correlation with their spontaneous, synchronized hair cycling, mice aged 8 and 14 weeks, with most of their hair follicles in telogen, exhibited strong CPS responses, whereas 4-, 11-, and 16-week-old mice with a predominance of anagen follicles in a large area of their integument exhibited lower responses. This suggests that the development of CPS is inhibited in mice with anagen hair follicles. Antigen-specific, T-cell receptor V beta 7+ suppressor T cells, which are recognized to down-regulate the CPS response to TCSA, were not generated in sensitized anagen mice. Culture supernatants of epidermal cells derived from mice with anagen hair follicles contained factor(s) that suppress in vivo the development of CPS. It was found that levels of mRNA for tumor necrosis factor alpha (TNF alpha) were markedly decreased in epidermal cells from early anagen to telogen mice, whereas message for IL-1 receptor antagonist (IL-1ra) was transcribed increasingly during this hair cycling. These findings suggest that altered keratinocyte production of these cytokines is involved in mediating the anagen-associated depression of CPS. PMID- 9225009 TI - CD26/dipeptidyl peptidase IV does not work as an adenosine deaminase-binding protein in rat cells. AB - In this paper, we describe further characterization of the membrane-associated molecule CD26/dipeptidyl peptidase IV (DPP IV), which is said to be adenosine deaminase-binding protein (ADA-bp) in humans, to clarify its association with ADA in rat immune cells. For this purpose, we used three types of rats: DPP IV+ rats; DPP IV- rats, which lack enzyme activity and immunological reactivity of DPP IV; and ADA- rats, which have reduced ADA activity due to continuous peritoneal injection of 2'-DCF, a potent inhibitor of ADA. ADA existed in the cells of DPP IV+ and DPP IV- rats, but it did not exist on the cell surface in either rat. ADA rats showed a decrease in ADA activity and in the number of immune cells, but no effect on DPP IV was observed. These data suggest that in rats, in contrast to humans, DPP IV does not exist as ADA-bp. PMID- 9225010 TI - In vitro production of antigen-specific T cells from unprimed mice: role of dexamethasone and anti-IL-10 antibodies. AB - We describe here a culture system for studying the development, in vitro, of antigen-specific CD4 T cells from unprimed mice. T cells from young mice are initially exposed to antigen, such as pigeon cytochrome C or keyhole limpet hemocyanin, in the presence of adherent accessory cells and then allowed to proliferate in the absence of antigen but in the presence of IL-2, 10(-8) M dexamethasone, and antibodies to IL-10. Proliferation and IL-2 production by T cells harvested from such expansion cultures are antigen-dependent but not antigen-specific and at different doses can be either stimulated or inhibited both by the priming antigen and by irrelevant proteins. Antigen-specific T cell reactions can be elicited by any of three modifications of the culture protocol: (a) absorption of nonspecific cells on accessory cell monolayers bearing irrelevant proteins; (b) increased doses of dexamethasone during the expansion phase; or (c) a second cycle of antigen activation and antigen-free expansion. These observations provide a foundation for further analysis of in vitro maturation of primary immune responses and suggest an important role for IL-10 and glucocorticoids in regulating the early stages of activation and proliferation by naive T cells. PMID- 9225011 TI - Influence of carcinogen binding by lactic acid-producing bacteria on tissue distribution and in vivo mutagenicity of dietary carcinogens. AB - The aims of this investigation were to determine whether viable cultures of lactic acid-producing organisms (LAB) can bind dietary carcinogens and to assess the consequences of binding for the absorption from the gut, distribution in the body and in vivo genotoxicity of ingested carcinogens. The carcinogens used in this study were ones known to be present in the human diet, namely benzo[a]pyrene (B(a)P, aflatoxin B1 (AFB1) and the cooked food carcinogens 2-amino-3-methyl-3H imidazo[4,5-f]quinoline (IQ), 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), 5-phenyl-2-amino-1 methylimidazo [4,5-f]pyridine (PhIP) and 3-amino-1-methyl-5H-pyrido[4,3-b]indole (Trp-P-2). They represent a range of structural types so that the specificity of any binding effects could be addressed. Of the carcinogens tested, B(a)P and Trp P-2 were bound most effectively by the two LAB strains Bifidobacterium longum and Lactobacillus acidophilus. AFB1 was poorly bound, while MeIQx, MeIQ, PhIP and IQ were bound to an intermediate degree. The extent of the binding of the heterocyclic amine carcinogens was dependent on the pH conditions during incubation and this effect was more apparent with B. longum than with L. acidophilus. Using the host-mediated assay (HMA), an in vivo bacterial mutation assay, it was demonstrated that the administration of bacterial cell suspensions of B. longum and L. acidophilus did not lead to a reduction in induced mutagenicity by MeIQ, MeIQx or Trp-P-2, detectable in the liver of treated mice compared with controls. The lack of a protective effect could not be attributed to a short period of contact between bacterial cells and mutagens, since similar results were obtained after preincubating bacteria and mutagens together at pH 5 for 50-60 min, to maximize the binding, before gavaging the mice. Lack of activity of B(a)P in the HMA prevented the determination of the effect of LAB on genotoxicity of the polycyclic aromatic hydrocarbon. However, it is clear from the radiolabel distribution study that the amount of the carcinogen entering the blood was not significantly reduced by B. longum administration. In addition, the amount of radiolabelled B(a)P that reached the target organs (liver, lungs and heart) was also not affected by the LAB administration. A similar lack of inhibitory effect of B. longum on blood concentration and accumulation in the liver of Trp-P-2 was apparent. The results of the present study suggest that although LAB may bind carcinogens in vitro, this does not lead to major changes in absorption and distribution of carcinogens in the body, or in their genotoxic activity in the liver. PMID- 9225012 TI - Analysis of the content of the diterpenes cafestol and kahweol in coffee brews. AB - The diterpenes cafestol and kahweol have been implicated as the components in boiled coffee responsible for its hypercholesterolaemic effects. These particular coffee constituents have also been shown to possess anticarcinogenic effects. A simple and sensitive reverse-phase HPLC method using solid-phase extraction has been developed for the analysis of cafestol and kahweol in coffee brews. This method was used to confirm that the method of coffee brewing is a major determinant of the cup content and hence level of consumption of these diterpenes. Scandinavian-style boiled coffee and Turkish-style coffee contained the highest amounts, equivalent to 7.2 and 5.3 mg cafestol per cup and 7.2 and 5.4 mg kahweol per cup, respectively. In contrast, instant and drip-filtered coffee brews contained negligible amounts of these diterpenes, and espresso coffee contained intermediate amounts, about 1 mg cafestol and 1 mg kahweol per cup. These findings provide an explanation for the hypercholesterolaemic effect previously observed for boiled coffee and Turkish-style coffee, and the lack of effect of instant or drip-filtered coffee brews. This methodology will be of value in more correctly assessing the human exposure to these diterpenes through the consumption of coffee, and hence the potential physiological effects of different brews. PMID- 9225013 TI - Polar and non-polar heterocyclic amines in cooked fish and meat products and their corresponding pan residues. AB - Fourteen cooked dishes with their corresponding pan residues were analysed for polar and non-polar heterocyclic amines using HPLC. The choice of foods, including beef, pork, poultry, game, fish, egg and sausages, was based on an investigation of an elderly population in Stockholm participating in an analytical epidemiological case-control study on cancer risks after intake of heterocyclic amines. The food items were prepared using normal household cooking practices, and to reflect the wide range of surface browning of the cooked dishes that would be encountered in this population, four cooking temperatures were used in the range 150-225 degrees C. For all food samples, the total amount of heterocyclic amines formed at 150 degrees C was less than 1 ng/g cooked product, and at 175 degrees C less than 2 ng/g. The highest concentrations of heterocyclic amines were detected in fillet of pork, reindeer meat and chicken breast fried at 200 and 225 degrees C and their corresponding pan residues. The total sum of 2 amino-3,8-dimethylimidazo-[4,5-f]quinoxaline, 2-amino-3,4,8-trimethylimidazo[4,5 f]quinoxaline and 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine was about 1 microgram per 100 g portion (including pan residues) for reindeer meat and chicken breast, and between 1.9 and 6.3 micrograms per 100-g portion for fillet of pork. PhIP was the most abundant heterocyclic amine, identified in 73 of 84 samples, and the highest concentration of PhIP, 32.0 ng/g, was found in the pan residue from fillet of pork cooked at 225 degrees C. The non-polar heterocyclic amines 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole and 3-amino-1-methyl-5H pyrido[4,3-b]indole were detected in the range of 0.5-7.4 ng/g in most foods cooked at 225 degrees C, and also in meat sauce prepared at 200 and 175 degrees C. The other heterocyclic amines tested for: 2-amino-3-methylimidazo-[4,5 f]quinoline, 2-amino-3,4-dimethylimidazo[4,5-f]quinoline, 2-amino-6-methyl-pyrido [1,2-a:3',2'-d]-imidazole and 2-aminodipyrido-[1,2-a:3',2'-d]imidazole, were present only at very low or non-detectable levels. The low recoveries of the amino-alpha-carbolines 2-amino-9H-pyrido[2,3-b]indole and 2-amino-3-methyl-9H pyrido[2,3-b]indole made it impossible to quantify them. However, the co mutagenic substances 1-methyl-9H-pyrido-[3,4-b]indole and 9H-pyrido[3,4-b]indole were detected at levels of about 1-30 ng/g in most of the dishes cooked at 200 and 225 degrees C. PMID- 9225014 TI - Sister chromatid exchanges induced in vitro and in vivo by an extract of black pepper. AB - Black pepper is a spice widely used in human food. The aim of this investigation was to determine whether an alcoholic extract of the mature berries of black pepper induced genotoxic damage in vivo and in vitro. The first aspect was evaluated in mouse bone marrow cells and the second one in human lymphocytes. In both cases the rate of sister chromatid exchange (SCE) and the replicative index were determined. For the in vivo assay, ip doses of 7.0, 14.0, 28.0 and 56.0 mg/kg body weight were tested, with the following results: (1) a significant increase of SCE frequency in all doses tested compared with the control level (the highest dose produced almost a duplication of the basal rate of SCEs); (2) a similar pattern with regard to cell proliferation kinetics at all doses tested, without significant differences between them. For the in vitro assay, doses of 25.0, 50.0, 75.0 and 100.0 micrograms/ml were tested, with the following results: (1) a significant increase in the frequency of SCEs at all doses tested; a linear regression analysis of the data produced a correlation coefficient of 0.98; (2) a significant reduction in the replicative index, at the two high doses. These results demonstrated that the extract of black pepper was genotoxic in both systems. PMID- 9225015 TI - Preliminary safety assessment of an arachidonic acid-enriched oil derived from Mortierella alpina: summary of toxicological data. AB - An arachidonic acid-enriched oil (AA-oil), derived from Mortierella alpina was subjected to a programme of studies to establish its preliminary safety for use in infant nutrition. This was addressed at two levels: (1) HPLC analysis of metabolites produced by the production strains at various conditions, and (2) an evaluation of the toxicity of the final product. The following studies were carried out on the AA-oil: gene mutation assays in bacteria and mammalian cells in vitro; chromosome aberration assays both in vitro and in vivo and acute and subacute (4-wk) oral toxicity in the rat. No known mycotoxins were produced by the production strains under the conditions tested. Further, the oil did not show mutagenic or clastogenic activity and the acute oral toxicity, expressed as the LD50 value, exceeded 20 ml/kg body weight, that is, 18.2 g/kg body weight. In the subacute oral toxicity study the AA-oil was tested as such and in combination with a docosahexaenoic-enriched oil (DHA-oil) derived from fish oil at a ratio of 2:1 (AA:DHA). This was done because high dose levels of AA may result in adverse effects; DHA can compensate for these effects. Furthermore, human milk contains both AA and DHA at a ratio of AA:DHA of 2 to 3:1. No obvious signs of toxicity were observed. Levels of phospholipids and triglycerides tended to be decreased in the highest dose groups. The no-observed-adverse-effect level of the AA-oil in the subacute 4-wk toxicity study was placed at the highest levels tested, namely 3000 mg AA-oil/kg body weight/day as such and in the combination of 3000 mg AA oil and 1500 mg DHA-oil/kg body weight/day. This corresponds to an intake of 1000 mg AA/kg body weight/day, which represents approximately 37 times the infant intake of AA in human milk. PMID- 9225016 TI - Safety evaluation of wild apricot oil. AB - Wild apricot, a variety of Prunus armeniaca, grows in the hilly regions of India. The seeds yield 27% of kernels. The potential availability of the kernels is 40,000 tons/year and these yield 47% of oil. The oil has 94% unsaturated fatty acids, rich in oleic and linoleic acids. Systemic effects and nutritional quality of wild apricot oil (WAO) were assessed in a 13-wk feeding study in weanling albino rats using a diet containing 10% WAO as the sole source of dietary fat. A similar diet containing groundnut oil (GNO) was used as the control. WAO did not manifest any toxic potential. The food consumption, growth rate and food efficiency ratio of rats fed WAO were similar to those fed GNO. The digestibility of this oil was found to be comparable to that of GNO. There were no macroscopic or microscopic lesions in any of the organs that could be ascribed to WAO incorporation in the diet. The results of this study indicate that WAO could be used for edible purposes without any overt toxic signs or symptoms. However a long-term study may be needed to confirm its innocuousness further. PMID- 9225017 TI - Metabolism of 14C-labelled sucrose esters of stearic acid in rats. AB - Rats were dosed by oral gavage (250 mg/kg) with compounds containing sucrose esterified in four, six or eight positions with stearic acid. For each compound, rats excreted greater than 95% of the dose in the faeces. The extent of absorption and metabolism of radioactivity was inversely related to the degree of esterification. For rats dosed with sucrose esters labelled in the fatty acid moieties, the degree of absorption of radioactivity was highest for the tetraesterified compound (5.9% of the dose). At 120 hr after dosing with this derivative, the highest concentrations of radioactivity, aside from tissues of the gastrointestinal tract, were found in fat (183 micrograms-equivalents/g tissue), lymph nodes (117 micrograms-equivalents/g tissue) and the liver (88 micrograms-equivalents/g tissue); appreciable radioactivity appeared in the blood (3.9 micrograms-equivalents/g tissue) and collected lymph (5.0-7.6% of the dose). For rats dosed with esters labelled in the sucrose moiety, the amounts of radioactivity absorbed were lower than after dosing with the corresponding sucrose derivatives labelled in the fatty acid moieties; the absorbed radioactivity was greatest following administration of the tetraesterified compound (3.0%). Relatively little radioactivity was found in tissue samples collected from these rats. These results are consistent with limited hydrolysis of the sucrose esters, presumably to sucrose and fatty acids, prior to intestinal absorption. PMID- 9225019 TI - Induction of micronuclei, DNA strand breaks and HPRT mutations in cultured Chinese hamster V79 cells by the phytoestrogen coumoestrol. AB - Coumoestrol (COUM), genistein (GEN) and daidzein (DAI) are major phytoestrogens present in numerous plants eaten by humans and food-producing animals. Little is known about the genotoxicity of these natural compounds. The effects of COUM, GEN and DAI were studied in cultured Chinese hamster V79 cells at various endpoints. None of the substances affected the cytoplasmic microtubule complex or the mitotic spindle. However, COUM and GEN but not DAI proved to be strong inducers of DNA strand breaks and micronuclei containing acentric fragments, as shown with antikinetochore antibodies. The clastogenicity of GEN may be due to its non intercalative inhibitory effect on topoisomerase II, whereas COUM may act through topoisomerase II inhibition and/or DNA intercalation. COUM was also a clear inducer of hypoxanthine guanine phosphoribosyltransferase (HPRT) mutations in V79 cells; GEN was only marginally active and DAI inactive at this endpoint. This is the first report on the clastogenicity and mutagenicity of COUM in mammalian cells. PMID- 9225018 TI - Assessment of the carcinogenicity of stevioside in F344 rats. AB - The carcinogenic potential of stevioside, a compound that is used as a sweetener for food and drink, was examined in F344 rats of both sexes. Stevioside was added to powdered diet at concentrations of 0 (control), 2.5 and 5%. The doses were selected on the basis of results from a 13-wk subchronic toxicity study and administered to groups of 50 male and 50 female rats ad lib. for 104 wk. All surviving rats were killed at wk 108. Body weight gains were slightly depressed in line with the dose of stevioside, in both sexes, and a significant decrease in the final survival rate was observed for the 5% treated males. Histopathologically, however, there was no significantly altered development of neoplastic or non-neoplastic lesions attributable to the stevioside treatment in any organ or tissue, except for a decreased incidence of mammary adenomas in females and a reduced severity of chronic nephropathy in males. It is concluded that stevioside is not carcinogenic in F344 rats under the experimental conditions described. PMID- 9225020 TI - Effect of nitrite on blood pressure in anaesthetized and free-moving rats. AB - The effect of nitrite on blood pressure and heart rate was studied in anaesthetized (non-telemetric method) and free-moving rats (biotelemetry system). In anaesthetized rats, NaNO2 (10-1000 mumol/kg), infused over 5 min, induced a dose-related decrease in blood pressure. The maximal decrease in mean arterial blood pressure (MAP), caused by 1000 mumol/kg NaNO2 and measured 15 min after infusion was 55.9 +/- 3.2% (n = 3). After NaNO2 infusion, in the plasma, rapid conversion of nitrite into nitrate was observed. However, sodium nitrate (NaNO3, 100 mumol/kg) did not decrease blood pressure and there was no conversion of nitrate into nitrite. Free-moving rats received KNO2 which was added to drinking water (36 mmol/litre) for a period of 3 days. KNO2 decreased the MAP and increased the heart rate during the rat's activity phase at night but not during their resting phase in the day. An equal concentration of potassium (KCl, 36 mmol/litre added to drinking water) for 3 days did not decrease blood pressure. It is concluded that nitrite decreases blood pressure in rats, which probably induces, by renin-angiotensin system activation, hypertrophy of the adrenal zona glomerulosa. PMID- 9225021 TI - Organochlorine pesticides in pasteurized milk and associated health risks. AB - The presence of organochlorine pesticides (alpha-HCH, beta-HCH, lindane, aldrin, dieldrin, heptachlor, heptachlor epoxide, chlordane and the isomers and metabolites of DDT) in Spanish pasteurized milk were investigated. 95% of the samples contained one of the isomers of the HCH group and 12.9% of them exceeded the maximum residue limit permitted by the European Union; six samples went over that limit for heptachlor epoxide and 74.63% of the samples contained chlordane at higher concentrations than those permitted by the legislation. None of the samples exceeded the limit for the DDT group. The mean concentrations detected for each pesticide were as follows (mg/kg): alpha-HCH = 0.015; beta-HCH = 0.039; lindane = 0.007; delta-HCH = 0.07; aldrin = 0.002; dieldrin = 0.028; chlordane = 0.101; heptachlor = 0.011; heptachlor epoxide = 0.021; o.p'-DDD = 0.016; p.p'-DDD = 0.009; p.p'-DDE = 0.045 (DDT + metabolites = 0.067). PMID- 9225022 TI - Assessment of cassia gum. AB - Cassia gum is approved for use in Europe by the Commission Directive (EEC No. E 499) and is listed in the Annex of the Council Directive (70/524/EEC) as a stabilizer (thickening and gelling agent) in the manufacture of canned pet foods (for cats and dogs). It is also approved for use in Japan and is listed as a food additive in The Ministry of Health and Welfare Announcement No. 160 (10 August 1995). A panel of experts in the areas of toxicology, pharmacology and food science was assembled to review the safety of cassia gum for use as a thickening agent in human and pet foods in the United States. The available data on cassia gum and structurally related gums demonstrate a lack of toxic effects in animals. This review is the basis for the consideration of cassia gum as generally recognized as safe (GRAS) under conditions of its intended use as a thickening agent in human and pet foods. PMID- 9225023 TI - Virus 'quasispecies': making a mountain out of a molehill? PMID- 9225024 TI - Differences in hepatitis C virus quasispecies composition between liver, peripheral blood mononuclear cells and plasma. AB - Hepatitis C virus (HCV) exists in vivo as a highly variable mixture of closely related genomes (quasispecies), but the pathogenetic significance of such heterogeneity is still largely unknown. To investigate this issue, we compared the composition of HCV quasispecies found in the liver, peripheral blood mononuclear cells (PBMC) and plasma of ten patients by single-strand conformation polymorphism analysis of the E2/NS1 region and sequencing of the variants detected. We found considerable quasispecies differences between the liver and PBMC in all the patients, involving variant numbers, relative quantities and relative electrophoretic mobilities, but no apparent tissue-specific trend. Genome variants present in the liver and/or PBMC were not detected in the corresponding plasma samples, while certain HCV variants were present only in plasma. No dominant amino acids or amino acid pattern characteristic of variants present solely in the PBMC were detected in the E2/NS1 region sequenced. PMID- 9225025 TI - Hepatitis C virus core protein induces hepatic steatosis in transgenic mice. AB - Hepatitis C virus (HCV) is a major cause of chronic hepatitis worldwide, which finally leads to development of hepatocellular carcinoma. Chronic hepatitis C is characterized by several histological features in the liver which discriminate it from other forms of hepatitis: bile duct damage, lymphoid follicles and steatosis (fatty change). Little is known, however, about the role of HCV or its viral proteins in the pathogenesis of hepatitis. Recently, the core protein of HCV has been suggested to have a transcriptional regulatory function, and thereby to be involved in inducing phenotypic changes in hepatocytes. To clarify whether or not the HCV core protein has an effect on pathological phenotypes in the liver, two independent transgenic mouse lines carrying the HCV core gene were established. These mice developed progressive hepatic steatosis, indicating that the HCV core protein plays a direct role in the development of hepatic steatosis, which characterizes hepatitis C. This transgenic mouse system would be a good animal model for the study of pathogenesis in human HCV infection. PMID- 9225026 TI - Discrimination of hepatitis G virus/GBV-C geographical variants by analysis of the 5' non-coding region. AB - We have investigated the ability of different subgenomic fragments to reproduce the phylogenetic relationships observed between six complete genome sequences of GBV-C/hepatitis G virus (HGV). While similar relationships were observed following analysis of part of the 5' non-coding region (5'NCR), for the coding region they were not accurately reproduced for some large fragments or for the majority of fragments of 300 or 600 nucleotides. Analysis of 5'NCR sequences from a large number of isolates, including newly obtained sequences from Pakistan, Zaire and Scotland, produced separate groupings of Asian, African and European/North American variants. These groupings are associated with specific polymorphisms in the 5'NCR, many of which were covariant and consistent with a proposed secondary structure for this region. The relatively low level of amino acid sequence variation observed between these geographically and phylogenetically defined groups of variants suggests that they are unlikely to display significant biological differences. PMID- 9225027 TI - Secondary structure of the 3'-untranslated region of yellow fever virus: implications for virulence, attenuation and vaccine development. AB - A genetic algorithm-based RNA secondary structure prediction was combined with comparative sequence analysis to construct models of folding for the distal 380 nucleotides of the 3'-untranslated region (3'-UTR) of yellow fever virus (YFV). A number of structural elements that are thermodynamically stable, conserved in shape, and confirmed by compensatory mutations were revealed. At the same time structural polymorphisms were observed among strains of YFV. These polymorphisms showed an association with virulence: all wild and pathogenic strains were likely to be folded in a significantly different way from vaccine strains with reduced virulence. Structural divergence was also found among vaccine strains, with 17DD, the most virulent in the mouse model, exhibiting an intermediate pattern of folding, combining structural features of both wild and vaccine strains. The observation of a strong association between secondary structure of the 3'-UTR and virulence of YFV may help elucidate the molecular mechanisms of virus attenuation and lead to new strategies of vaccine development directed towards rational modification of secondary structure of the 3'-UTR. PMID- 9225028 TI - Sequence analysis of the avirulent, demyelinating A7 strain of Semliki Forest virus. AB - The nonstructural region of the genome of the avirulent A7 strain of Semliki Forest virus (SFV) has been sequenced, so that the complete nucleotide sequence is available. Compared to the virulent SFV4 strain (produced from the infectious clone pSP6-SFV4), A7 contains 226 nucleotide changes in the translated region, which result in 47 amino acid changes. The 5' nontranslated region has two nucleotide changes, and the 3' nontranslated region is longer in A7 than SFV4, and contains divergent and repeated sequences. Chimeras containing SFV4 and A7 sequences and an infectious clone of A7, pSP6-CA7, were constructed. The virulence of these was tested by intraperitoneal and intranasal infection of adult BALB/c mice. It was shown that determination of the avirulent phenotype of A7 was polygenic, and required the additive effect of sequences from both the structural and non-structural regions of the SFV genome. PMID- 9225029 TI - Death mechanisms in cultured cells infected by Semliki Forest virus. AB - We have investigated the induction of cell death in cultured cells by the virulent SFV4 and avirulent A7 strains of Semliki Forest virus (SFV). In BHK cells, death occurred by a typical apoptotic mechanism, as did the death of oligodendrocytes in glial cell cultures. For cerebellar neuron cultures, virus induced death was due to necrosis. Although the SFV4 and A7 strains did not differ in the mechanism of induction of cell death, the virulent SFV4 strain did multiply to a higher titre in cultured neurons than the avirulent A7 strain. This is consistent with previous animal studies which indicate that the virulence of SFV strains is controlled by rapidity of multiplication in the CNS, leading to a lethal threshold of damage, rather than differential cell tropism or cell death mechanisms. The immune-mediated demyelination induced by avirulent strains may be triggered by apoptosis of oligodendrocytes, the consequences of which are obscured by death for virulent strains. PMID- 9225030 TI - Molecular cloning of the hepatitis A virus receptor from a simian cell line. AB - Using a eukaryotic expression system in combination with a monoclonal antibody (MAb) capable of blocking hepatitis A virus (HAV) adsorption, a cDNA clone was selected from a library of S.la/Ve-1 cells, a cell line that is highly susceptible to the virus. Sequence analysis of the cDNA revealed a single open reading frame that encoded a protein consisting of 460 amino acids. The deduced primary structure of the protein included a signal sequence, a transmembrane domain, four sites for N-linked glycosylation, cysteine residues attributable to an immunoglobulin domain and threonine clusters characteristic of mucin-like protein. By employing a vaccinia virus expression vector, the cDNA was expressed in HeLa cells where it induced marked HAV attachment which was specifically blocked by the MAb. The cDNA obtained was thus assumed to encode a functional receptor for HAV. PMID- 9225031 TI - The complete Mokola virus genome sequence: structure of the RNA-dependent RNA polymerase. AB - The genome sequence of the rabies-related virus Mokola virus (genus Lyssavirus) has been completed by sequencing the L gene, which consists of 6384 nucleotides encoding a 2127 amino acid polymerase. Alignment of the Mokola virus L protein with other polymerases from the virus order Mononegavirales defined three domains: a divergent NH2-terminal domain, a highly conserved central domain carrying most of the functional motifs and a COOH-terminal domain with alternating conserved and divergent regions. A statistical study outlined the stringency of conservation of glycine, acidic (D, E) and basic (K, R, H) amino acids in polymerases, particularly as key residues of the conserved motifs. PMID- 9225032 TI - Class I-restricted CTL induction by mucosal immunization with naked DNA encoding measles virus haemagglutinin. AB - We have investigated the class I-restricted CTL response specific for measles virus haemagglutinin (HA) in the spleens of mice immunized by various mucosal routes with a DNA plasmid carrying the HA gene (pV1j-HA). A single immunization with recombinant DNA injected in the buccal mucosa induced an HA-specific CTL response. Similarly, nasal immunization with the DNA vaccine induced primary CTLs against measles virus HA. Booster immunization did not enhance the CTL activity. Oral or intrajejunal immunization with the plasmid induced a CTL response of lower magnitude. However, this could be potentiated by co-administration of the mucosal adjuvant cholera toxin or cationic lipids (DOTAP). These data show that a CTL response can be generated by mucosal vaccination using DNA vaccines. PMID- 9225033 TI - Sequence analysis of the nucleocapsid gene of measles virus isolates from South Africa identifies a new genotype. AB - Sequence analysis was performed on 20 measles virus (MV) isolates from South Africa, five of which were obtained between 1986 and 1989 and 15 isolates collected during the 1994/95 measles season. A 590 bp fragment of the carboxyl terminus of the nucleocapsid (N) was amplified by PCR and subjected to sequence and phylogenetic analysis. Comparison of the South African MV strains with those previously described revealed that at least two distinct groups of wild-type (wt) MV exist, one of which has been circulating since 1986. The major genotype (I) was represented by the more recent isolates which showed three characteristic amino acid substitutions. Furthermore, three vaccine-like viruses with sequences very similar to the Edmonston wt strain were identified. Phylogenetic analysis of 100 MV strains allowed the assignment of new definitions for MV genotypes and subgroups. Employing these definitions, the majority of South African isolates analysed here formed a new genotype. PMID- 9225034 TI - Influenza virus M1 protein binds to RNA through its nuclear localization signal. AB - The RNA-binding activity of influenza A virus M1 protein was studied by cross linking the protein to viral RNA followed by sequence analysis of the oligoribonucleotide bound to the protein as well as sequence analysis of the M1 peptide bound to the RNA. M1 was found to bind to RNA without any RNA sequence specificity, as verified in a series of filter-binding experiments using a large variety of nucleic acids including DNA. The peptide sequence that bound to the RNA was the RKLKR nuclear localization signal of M1. Site-specific mutagenesis of recombinant M1 showed that most of the basic residues in that region had to be mutated in order to inhibit RNA-binding. We also constructed an M1 mutant that no longer bound to RNA but which was still able to inhibit the in vitro transcription activity of isolated viral ribonucleoprotein, albeit to a lower extent. Mutation of the zinc-binding sequence had no effect on RNA-binding or transcription-inhibition activity. PMID- 9225035 TI - Role of gamma delta TCR+ lymphocytes in the augmented resistance of trehalose 6,6'-dimycolate-treated mice to influenza virus infection. AB - Trehalose 6,6'-dimycolate (TDM), an immunomodulator, potentiates non-specific resistance in mice to influenza virus infection. When mice were injected intravenously with TDM, the striking proliferation of a minority of T-lymphocytes bearing gamma/delta T-cell receptors (gamma delta T-cells) that accumulated in granulomatous lungs was thought to be associated with the maintenance of acquired resistance to lethal influenza virus infection. To clarify the cellular basis of the defence against influenza virus, mice were depleted of gamma delta T-cells, alpha/beta (alpha beta) T-cells, or natural killer (NK) cells by in vivo administration of corresponding antibodies prior to influenza virus infection. The depletion of gamma delta T-cells significantly abrogated the augmented resistance of TDM-treated mice to infection, as did depletion of either alpha beta T-cells or NK cells. To gain insight into the functional ability of gamma delta T-cells, we evaluated the cytotoxic activity of this T-cell subset against a panel of target cell lines that were stably transfected with the influenza virus haemagglutinin (HA) gene from A/PR/8/34(H1N1) and A/Aichi/2/68(H3N2) strains. The gamma delta T-cells from TDM-treated mice showed profound cytotoxicity against the target cells expressing HA of either the H1 or H3 subtype, in a non-major histocompatibility complex-restricted manner. Taken together, these results indicate that gamma delta T-cells play a non-specific role, in conjunction with alpha beta T-cells and NK cells, in protecting mice against influenza virus infection, and that the recognition and destruction of HA expressing target cells by the activated gamma delta T-cells is one of the steps involved in this anti-influenza virus immunosurveillance. PMID- 9225036 TI - Molecular characterization of attenuated Junin virus strains. AB - The Junin virus strain Candid #1 was developed as a live attenuated vaccine for Argentine haemorrhagic fever. In this paper we report the nucleotide sequences of S RNA of Candid #1 and its more virulent ancestors XJ#44 and XJ (prototype). Their relationship to Junin virus wild-type MC2 strain and other closely and distantly related arenaviruses was also examined. Comparisons of the nucleotide and amino acid sequences of N and GPC genes from Candid #1 and its progenitor strains revealed some changes that are unique to the vaccine strain. These changes could be provisionally associated with the attenuated phenotype. PMID- 9225037 TI - African horsesickness virus VP7 sub-unit vaccine protects mice against a lethal, heterologous serotype challenge. AB - An established mouse model was used to evaluate the effectiveness of the major outer core protein of African horsesickness virus (AHSV), VP7, as a subunit vaccine. Adult female BALB/c mice were immunized with VP7 crystals purified from BHK cells infected with AHSV serotype 9 (AHSV-9), using three inoculations in Freund's adjuvant. Eighty to one hundred per cent of the immunized mice were protected against a heterologous challenge with a known lethal dose of AHSV-7. The protected immunized mice did not develop any clinical signs characteristic of virulent AHSV infection in this model during the study. In contrast, 80-100% mortality was observed in the non-immunized mice that received the same challenge virus. Subsequent studies indicated that a single inoculation of 1.5 micrograms purified AHSV VP7 in Freund's complete adjuvant was sufficient to protect at least 90% of mice from AHSV-7 challenge. If the antigen was presented in the absence of Freund's complete adjuvant, 70% of the mice were still protected by one inoculation of VP7 crystals. Titres of circulating antibody against AHSV VP7, determined by competitive ELISA, did not appear to correlate with protection and passive antibody transfer from immunized BALB/c mice failed to protect syngeneic recipients from AHSV-7 challenge. Therefore, the observed protection is unlikely to be due to an antibody-mediated immune response. The number of viraemic mice and the duration of viraemia post-challenge was significantly reduced in vaccinated mice compared to non-vaccinated controls. However, the levels of viraemia were similar. PMID- 9225038 TI - VP7: an attachment protein of bluetongue virus for cellular receptors in Culicoides variipennis. AB - The importance of VP7 of bluetongue virus (BTV) in the binding of BTV to membrane proteins of the BTV vector Culicoides variipennis was investigated. Core BTV particles, prepared from whole viruses, lacked outer proteins VP2 and VP5 and had VP7 exposed. More core particles and whole viruses bound to membrane preparations of adults of C. variipennis and KC cells, which were cultured from this vector insect, than to membrane preparations of Manduca sexta larvae. More core particles than whole viruses bound to membrane preparations of adults of C. variipennis and KC cells. Polyclonal anti-idiotypic antibodies (anti-Id), which were made against an antigen-combining region of an anti-BTV-10 VP7 antibody and functionally mimicked VP7, bound more to the membrane preparations of adults of C. variipennis and KC cells, and less to cytosol preparations. In Western overalay analysis, the Culicoides plasma membrane preparation reduced binding of an anti-VP7 monoclonal antibody to VP7. Whole and core BTV particles and the anti Id bound to a membrane protein with a molecular mass of 23 kDa that was present predominantly in membrane preparations of adults of C. variipennis and KC cells. This protein was present in much lower concentrations in membrane preparations of C6/36 and DM-2 insect cells. PMID- 9225040 TI - Efficient herpes simplex virus type 1 (HSV-1) capsid formation directed by the varicella-zoster virus scaffolding protein requires the carboxy-terminal sequences from the HSV-1 homologue. AB - The scaffolding protein and associated protease of the human herpesvirus varicella-zoster virus (VZV), encoded by genes 33.5 and 33 respectively, were synthesized in insect cells using a baculovirus expression system. The expressed 33.5 product formed numerous long, flexible, hollow rods, and in this respect different from the herpes simplex virus type 1 (HSV-1) homologue which forms large aggregates consisting mainly of fibrous material interspersed with scaffold like particles. Removal of 27 amino acids from the carboxy terminus of the VZV scaffolding protein by the gene 33 protease or expression of the cleaved product did not result in any discernible change in the morphology of the scaffolding protein. Again, this was in marked contrast to the situation in HSV-1 where removal of the 25 carboxy-terminal amino acids from the scaffolding protein by the associated protease or expression of VP22a results in the formation of large numbers of scaffold-like particles. Despite these differences, when cells were multiply infected with baculoviruses expressing the HSV-1 capsid shell proteins and the VZV scaffolding protein complete capsids were observed, suggesting that the VZV protein could act as a scaffold for the assembly of the HSV-1 capsid shell. The efficiency of capsid assembly was increased substantially by exchanging the 23 carboxy-terminal amino acids of the VZV scaffolding protein for the corresponding 22 carboxy-terminal amino acids of the HSV-1 homologue, supporting previous work which showed that this region was critical for the formation of intact capsids. PMID- 9225041 TI - Nuclear translocation of mutagenized forms of human cytomegalovirus glycoprotein B (gpUL55). AB - To define structural elements involved in translocation of human cytomegalovirus (HCMV) glycoprotein B (gB) to the inner nuclear membrane (INM) compartment, mutagenized gB derivatives with deletions of the potential membrane anchor domains or of portions of the cytoplasmic tail were stably expressed in human astrocytoma cells. Subcellular localization examined by immunofluorescence and cell fractionation suggested that all gB derivatives reached the INM; however, reduced amounts were found after deletion of the extreme carboxy terminus [amino acids 856-906; gB(Del3)]. Pulse-chase analysis revealed accumulation in nuclear fractions of all gB derivatives during the chase, except for gB(Del3), which exhibited impaired nuclear retention. A carboxy-terminal nucleoplasmin-like signal localized within the respective deletion may thus be involved in nuclear transport and retention of HCMV gB. Immunoprecipitation after 32P-radiolabelling of the gB transfectants verified that the gB molecule is phosphorylated at a carboxy-terminal consensus motif for casein kinase II. PMID- 9225039 TI - Differential T cell response induced by certain recombinant oligopeptides of herpes simplex virus glycoprotein B in mice. AB - Much attention is presently focused on the quality of the immune response produced by helper T or regulatory cells because of its implications for vaccine development and immunomodulation. Glycoprotein B (gB) of herpes simplex virus (HSV) has been shown to induce a protective T cell response. To further characterize the nature of the T cell response, oligopeptides were expressed from the open reading frame of gB from HSV-2 (gB-2) as fusion proteins with beta galactosidase (GZ) in E. coli. After immunopurification using an anti-GZ affinity column, oligopeptides p59 and p65, spanning amino acid residues 339-394 and 424 484 of gB-2 respectively, were examined for immunogenic response by delayed type hypersensitivity (DTH) in vivo and for antigenic response by T cell proliferation in vitro. p59 but not p65 was able to prime for both DTH and proliferative T cell response to whole HSV-2 and protect against challenge infection. However, when mice were pretreated with cyclophosphamide, p65 primed for a strong DTH response to a level similar to that induced by p59 in mice either pretreated or not treated with cyclophosphamide. This suggests that p65 contains epitopes capable of inducing both DTH and immunosuppression. Thus, when mice were primed with p65 before immunizing with HSV-2, their in vitro HSV-specific proliferative response was suppressed. Therefore, p59 is a good immunogen able to induce significant, though incomplete, protection. It could be considered for inclusion in a cocktail of subunit vaccines against HSV-2 whereas p65 or parts thereof should be excluded for this purpose. PMID- 9225042 TI - Comparison of the human versus murine cytomegalovirus immediate early gene promoters for transgene expression by adenoviral vectors. AB - We have developed a number of replication defective adenoviral (Ad) vectors which express transgenes under the control of the human cytomegalovirus (HCMV) immediate early (IE) gene promoter. The orientation of the expression cassette replacing E1 in the vector backbone had a significant effect on the level of transgene expression, with vectors containing expression cassettes directed towards the right end of the Ad genome expressing 7-fold higher levels of beta galactosidase (beta-gal) than those with inserts in the opposite orientation. Murine cells infected with any of several different Ad vectors in which transgene expression was under the control of the HCMV IE promoter produced 10-100-fold less transgene product (such as beta-gal) than similarly infected human cells. Replacing the HCMV IE promoter with the murine CMV (MCMV) IE promoter resulted in an increase in the levels of beta-gal produced in murine and rat cells by approximately 5-30-fold compared to levels obtained with the HCMV IE promoter, and levels produced in human cells were the same or greater using the MCMV IE promoter compared to the HCMV IE promoter. Similar results were obtained using a luciferase reporter gene. The MCMV IE promoter, therefore, was able to drive high levels of expression without the pronounced species preferences observed for the HCMV IE promoter. The MCMV IE promoter also directed high levels of expression in vivo, suggesting that Ad vectors carrying the MCMV IE promoter may be more effective than those with the HCMV IE promoter for transgene expression in animal models. PMID- 9225043 TI - Sequence variations in EBNA-1 may dictate restriction of tissue distribution of Epstein-Barr virus in normal and tumour cells. AB - In seropositive individuals Epstein-Barr virus (EBV) establishes a virus reservoir in peripheral blood lymphocytes (PBLs). Transmission from one individual to another occurs via saliva due to a lytic (virion productive) phase of infection in the oropharynx. EBNA-1 is responsible for maintaining viral episomes in the host cell and could, therefore, also affect the persistence of the virus in different cell lineages. Based on sequence analysis of EBNA-1 we now demonstrate that (i) in addition to the prototype EBNA-1 (identical to the B95.8 virus EBNA-1), EBV in normal individuals encompasses multiple EBNA-1 subtypes, both in PBLs and in oral secretions; (ii) although EBV with prototype EBNA-1 is the predominant virus in normal individuals, it is very rarely associated with either nasopharyngeal carcinoma (NPC) or Burkitt's lymphoma (BL); (iii) EBV with an EBNA-1 subtype (V-val) frequently associated with NPC is also selectively detected in oral secretions and not in PBLs; (iv) EBV with the EBNA-1 subtype V pro is restricted to PBLs, while a mutated version of this subtype is present in BL, but not in NPC. These findings suggest that the variations in EBNA-1 may be relevant to the ability of EBV to persist in different cell types, and hence relevant to its oncogenic potential. PMID- 9225044 TI - Both A type and B type Epstein-Barr virus nuclear antigen 6 interact with RBP-2N. AB - Using the yeast two-hybrid system, Epstein-Barr virus nuclear antigen 6A (EBNA6A) was found to interact with the RBP-2N isoform of RBP-J kappa. The interaction of EBNA6A and EBNA6B with RBP-2N was compared and the results indicated that EBNA6B was less efficient at interacting with RBP-2N than was EBNA6A. Deletion mutation analysis of EBNA6A identified a region involved in the interaction with RBP-2N, while analysis of RBP-2N identified a domain which interacts with EBNA6A. The region of RBP-2N to which EBNA6A binds has previously been shown to interact with EBNA2. PMID- 9225045 TI - Murine gammaherpesvirus 68 encodes tRNA-like sequences which are expressed during latency. AB - Murine gammaherpesvirus 68 (MHV-68) is a virus of wild rodents and is a convenient small animal model for studies of gammaherpesvirus pathogenesis. We have sequenced 6162 bp at the left end of the MHV-68 genome and identified two unique open reading frames (ORFs) (ORF2 and ORF3) and an ORF (ORF1) which displays similarity to poxvirus members of the serpin family. Interspersed with the ORFs is a family of eight novel tRNA-like sequences sharing tRNA-like predicted secondary structures and RNA polymerase III promoter elements. These sequences are expressed to high levels during lytic infection and are processed into mature tRNAs with post-transcriptionally added 3' CCA termini, indicating their recognition as tRNAs by cellular machinery. Acidic Northern analysis of four tRNAs tested has demonstrated that they are not aminoacylated by aminoacyl tRNA synthetases present in the infected cell. Thus, it is currently unclear what biological function these uncharged viral tRNA-like sequences may fulfil. In situ hybridization analysis has shown that in addition to being expressed within productively infected tissues during acute stages of infection, the tRNA-like sequences are abundantly expressed within splenic germinal centres of latently infected mice. Therefore, the MHV-68 viral tRNAs represent a marker for latent infection and constitute the first report of tRNA-like sequences encoded by a virus of eukaryotes. PMID- 9225046 TI - Specificity of human cytotoxic T lymphocytes induced by a human papillomavirus type 16 E7-derived peptide. AB - In order to establish tumour-specific cytotoxic T lymphocyte (CTL) cell lines, T cells from a human papillomavirus (HPV) type 16-positive patient with a cervical carcinoma in situ and from a healthy volunteer were stimulated in vitro with autologous dendritic cells loaded with peptides derived from the viral transforming proteins E6 and E7 and corresponding to potential HLA-A*0201 restricted T cell epitopes. From each donor a small number of low-affinity CTL lines against the peptide E7/86-93 was obtained, which specifically lysed HLA A*0201-expressing B-lymphocytes (cell line 721) loaded with this peptide. Cytotoxicity was also observed against two HLA-A*0201-E7-positive epithelial cell lines, the cervical carcinoma cell line CaSki and the HPV-16-immortalized foreskin-keratinocyte cell line HPK IA. However, since none of the CTL recognized both cell lines, and E7-expressing 721 transfectants were never lysed, it was concluded that the reactivity against CaSki and HPK IA cells was due to cross reactivity on allogeneic HLA molecules rather than to E7 recognition, which emphasizes that the specificity of tumour cell lysis by peptide-induced CTL has to be interpreted with caution. PMID- 9225047 TI - Tissue culture adaptation of natural isolates of simian virus 40: changes occur in viral regulatory region but not in carboxy-terminal domain of large T-antigen. AB - The regulatory region of natural isolates of simian virus 40 (SV40) is different from that of laboratory-adapted strains of the virus. The latter have a nucleotide sequence duplication within the enhancer region which varies slightly with each strain, whereas the duplication is lacking in fresh isolates of SV40, which contain an 'archetypal' regulatory region. Many isolates also display nucleotide differences in the DNA encoding the carboxy terminus of large tumour antigen (T-ag). To determine whether genetic changes in these two regions of the SV40 genome were detectable during laboratory adaptation and long-term passage, low-passage virus stocks of two laboratory strains which had detailed passage histories spanning more than 25 years (Baylor strain and VA45-54) were analysed using PCR, cloning and sequencing assays. Both laboratory and archetypal regulatory regions were present in low-passage stocks. Following duplication in the regulatory region, no additional changes were detectable. The variable region at the T-ag carboxy terminus did not undergo any change with tissue culture passage and may serve as a useful site for taxonomic classification of different strains of SV40. Cloned genomes containing single or duplicated enhancers derived from both SV40 strains were viable in CV-1 cells. Attempts to induce regulatory region duplications by 14 serial passages of SV40 archetypal strains in monkey cells were not successful. The results are compatible with tissue culture adaptation of SV40, reflecting either selection of a rare variant pre-existing in the original sample or generation of a rare regulatory region duplication in infected cells. PMID- 9225048 TI - Sequence heterogeneity of heron hepatitis B virus genomes determined by full length DNA amplification and direct sequencing reveals novel and unique features. AB - So far, only a single heron hepatitis B virus genome (HHBV-4) has been cloned and sequenced. Therefore, neither the significance of its sequence divergence from other avian hepadnaviruses nor the sequence variability of HHBV genomes in general are known. Here we have analysed the sequence heterogeneity of HHBV genome populations in several sera from naturally infected herons. A highly sensitive PCR method for full-length HHBV genome amplification was established which allowed direct sequencing of entire HHBV populations without prior cloning. Sequences of HHBV genomes from four sera were thus obtained which differed from those of HHBV-4 by up to 7%. Some of the divergent nucleotides and the corresponding amino acids of the predicted viral proteins were conserved in all four new HHBV isolates and varied only in HHBV-4. This indicates that the HHBV-4 genome is not in all aspects representative of this class of viruses. Interestingly, a highly conserved ORF upstream of the C-gene present in a position analogous to that of the mammalian hepadnavirus X-gene became apparent in all HHBV genomes. In contrast to the duck hepadnaviruses, the small (sAg-S) instead of the largest (sAg-L) envelope protein of all HHBVs has a myristylation site. These data confirm the significant sequence divergence of HHBV from other avian hepadnaviruses. Moreover, they show that HHBV has low sequence variability and indicate two new and unique features not evident in other avihepadnaviruses: an additional, highly conserved gene and potential myristylation of the sAg-S instead of the sAg-L envelope protein. PMID- 9225049 TI - A unique segment of the hepatitis B virus group A genotype identified in isolates from South Africa. AB - The preS2/S genes of hepatitis B virus isolated from 29 acutely or chronically infected individuals in the Gauteng province of South Africa were sequenced. Phylogenetic analysis of these sequences in comparison with global isolates from the GenBank database showed that 24 sequences clustered with genotypic group A, three with genotypic group D and one each with genotypic groups B and C. Group A isolates had greater identity with groups D (variation of 6.6%) and E (6.8%) than with the Eastern groups B (7.4%) and C (8.1%) and were most different from group F (11.0%). Of the South African group A specimens, 59.1% clustered with two global sequences to form a discrete segment which we have called subgroup A. The amino acid differences that set these isolates apart from the rest of group A tended to cluster in the preS2 region (amino acids 7, 10, 32, 35, 47, 48, 53 and 54), with a few changes occurring in the major surface antigen (amino acid sites 207 and 209). Analysis of isolates showed that there was a 9-fold higher prevalence of the ay determinant in South Africa than previously reported. PMID- 9225050 TI - Structure and genomic organization of a novel human endogenous retrovirus family: HERV-K (HML-6). AB - Prototypic elements of a novel human endogenous retrovirus (HERV) family were identified and cloned from a human genomic library by the use of a pol fragment, HML-6, related to type A and type B retroviruses and class II HERVs. Out of 39 polhybridizing clones, five contained structures of full-length retroviral proviruses, with regions showing similarity to gag, pol and env, flanked by long terminal repeats (LTRs). Restriction mapping and partial sequence analysis of each full-length clone revealed few conserved restriction sites among HML-6 genomes, and about 20% sequence divergence over the reverse transcriptase region sequenced, suggesting that HML-6 constitutes a heterogeneous, but distinct family of elements belonging to the HERV-K superfamily. Sequence analysis of two clones, HML-6p and HML-6.17, revealed a lysine (K) tRNA UUU primer-binding site, and 40 68% nucleotide sequence similarity to LTR, gag, pro, pol and env regions of type B retroviruses and class II HERVs. HERV-K (HML-6) elements are present at about 30-40 copies per haploid genome. The HML-6 LTRs contain putative progesterone responsive elements, which may be involved in the regulation of HML-6 expression. Furthermore, there are about 50 additional solitary HML-6 LTRs per haploid genome. Such LTRs were integrated within the pol region of two clones belonging to the same HML-6 family, indicating that some site preference may be involved in HERV integration. PMID- 9225051 TI - Transfer of endoplasmic reticulum and Golgi retention signals to human immunodeficiency virus type 1 gp160 inhibits intracellular transport and proteolytic processing of viral glycoprotein but does not influence the cellular site of virus particle budding. AB - In this study, specific signals known to mediate endoplasmic reticulum or Golgi localization of transmembrane proteins have been transferred to the human immunodeficiency virus type 1 (HIV-1) env gene product. The intracellularly retained recombinant glycoproteins were not proteolytically processed to gp120 and gp41, which is further evidence that this process occurs at a later stage in the transport pathway, presumably within or near the trans-Golgi network. Since the subcellular localization of the viral glycoproteins of enveloped viruses can be one of the factors determining the cellular site of particle assembly and release, experiments were performed to determine if this property was altered by coexpression of the recombinant HIV-1 glycoproteins. When wild-type virus was compared to mutant virus encoding the intracellularly retained glycoproteins, the extent of HIV-1 particle release into the extracellular medium remained unaffected, and electron-microscopic analysis did not reveal any significant alteration in the cellular sites of particle assembly and budding. Thus, in COS-7 cells, altered subcellular localization of the viral glycoprotein does not exert a dominant influence on the assembly site of the HIV-1 particle. PMID- 9225052 TI - A protoplast system for studying tomato spotted wilt virus infection. AB - A plant protoplast system for studying tomato spotted wilt tospovirus (TSWV) infection was established and tested. Using polyethylene glycol-mediated inoculation with highly infectious TSWV particles, generally 50% or more of Nicotiana rustica protoplasts were infected. In these cells viral RNA and viral protein synthesis became detectable at 16 h post-inoculation (p.i.) and continued at least until 90 h p.i. Both the structural viral proteins [nucleoprotein (N) and the envelope glycoproteins G1 and G2] and the nonstructural viral proteins NSs and NSm accumulated to amounts sufficient for detection and immunocytological analysis. Local lesion tests on petunia leaves and electron microscopical analysis confirmed the production of mature, infectious virus particles, underlining the conclusion that a full infection cycle was completed in this system. Upon inoculation of Vigna unguiculata (cowpea) protoplasts with TSWV particles, comparable proportions of infected cells and amounts of NSs, NSm and N protein were obtained, but much lower amounts of viral glycoproteins were detected than in N. rustica protoplasts, and progeny virus particles were less abundant. With the N. rustica-based protoplast system, a powerful synchronized single-cell infection system has now become available for more precise in vivo studies of the processes occurring during tospovirus infection. PMID- 9225053 TI - A cDNA clone from a defective RNA of citrus tristeza virus is infective in the presence of the helper virus. AB - A naturally occurring defective RNA of 2379 nt (D2.3) from the VT strain of citrus tristeza closterovirus (CTV) was cloned and sequenced. The D2.3 RNA is a fusion of two regions of 1521 and 858 nt from the 5' and 3' ends of the CTV genome, respectively. A cDNA clone of D2.3 RNA was tagged by the insertion of a 0.47 kb chimeric DNA fragment and the recombinant cDNA was inserted downstream of the cauliflower mosaic virus 35S promoter. The resulting construct was bombarded into CTV-infected tissue, which was then grafted onto virus-free plants. The presence of recombinant RNA in systemically infected leaves was demonstrated by RT-PCR. Sequencing the RT-PCR products synthesized from double-stranded RNA confirmed the presence of the chimeric segment used for tagging. This is the first report of an infectious cDNA molecule derived from CTV D-RNA. PMID- 9225054 TI - Transgenic accumulation of two plant virus coat proteins on a single self processing polypeptide. AB - An expression cassette based on the highly specific tobacco etch potyvirus (TEV) nuclear inclusion (NIa) proteinase has been developed to produce multiple proteins through the translation of a single self-processing polypeptide. Gene constructs encoding TEV NIa, the tobacco mosaic tobamovirus (TMV) coat protein (CP) and the soybean mosaic potyvirus (SMV) CP were used to develop transgenic tobacco plants. Proper processing of the multifunctional polypeptide was demonstrated, leading to accumulation of separate proteins in planta. Moreover, the viral genes expressed in this way were biologically active and conferred pathogen-derived protection to TMV, TEV and potato potyvirus Y (PVY). Transgenic plants were also derived from gene constructs in which the NIa cleavage site was mutated, resulting in the accumulation of the non-processed polyprotein, as predicted. Although transgenic proteins accumulated in low amounts in all the plant lines analysed, accumulation of the mutant non-processed protein form was greatly increased in plants following infection with TEV, but not TMV, apparently as a consequence of protein stabilization. PMID- 9225055 TI - Detection of potato mop-top virus capsid readthrough protein in virus particles. AB - Potato mop-top furovirus (PMTV) RNA 3 encodes the 20 kDa coat protein and a larger readthrough protein of 67 kDa. The readthrough protein is expressed by suppression of the amber stop codon which terminates the coat protein gene. A 21 kDa C-terminal fragment of the readthrough protein was doned, fused to glutathione S-transferase and expressed in E. coli. An antiserum prepared against purified fusion protein was used in ELISA to detect the readthrough protein in extracts of PMTV-infected leaves. Immunogold labelling studies showed that the readthrough protein was located near one extremity of some of the virus particles. PMID- 9225056 TI - Nucleotide sequence of a new bipartite geminivirus isolated from the common weed Sida rhombifolia in Costa Rica. AB - The nucleotide sequence of infectious clones of a geminivirus from Costa Rica that infects Sida rhombifolia was determined. Sida golden mosaic virus (SiGMV-Co) has a bipartite genome (DNAs A and B). Computer analysis showed that the bipartite genome of SiGMV-Co resembles that of other whitefly-transmitted geminiviruses. The DNA A (2605 nt) and DNA B (2587 nt) components have little sequence homology other than within the common region (CR). Analysis of DNAs A and B showed that SiGMV-Co is closely related to bean dwarf mosaic virus (BDMV). SiGMV-Co was introduced via agroinoculation into seven plant species, including tomato and bean. PMID- 9225057 TI - Efficient whitefly transmission of African cassava mosaic geminivirus requires sequences from both genomic components. AB - Clones of two subgroup III geminiviruses, the common strain of tomato golden mosaic virus (csTGMV) and African cassava mosaic virus originating from Kenya (ACMV-K), were shown to be non-transmissible by whitefiles. Lack of transmissibility of cloned ACMV-K was investigated by exchanging genomic components with a whitefly-transmissible ACMV isolate from Nigeria (ACMV-NOg). Neither pseudorecombinant was transmissible, indicating that defects in both genomic components contributed to the lack of transmissibility. Analysis of the acquisition of the pseudorecombinats by Bemisia tabaci indicated that accumulation of virus within the insect was DNA B dependent. Return of virus to plants was determined by DNA A, although the coat protein was essential for acquisition. Repeated passaging of both the wild strain of ACMV-NOg and the cloned virus led to loss of insect transmissibility of the wild isolate but not the cloned virus. Products encoded on both genomic components are required for transmission of bipartite geminiviruses by B. tabaci. PMID- 9225058 TI - Nicking and joining activity of banana bunchy top virus replication protein in vitro. AB - The major open reading frame of banana bunchy top virus (BBTV) DNA-1 encodes a putative replication initiation protein (Rep). In vitro, a fusion protein of BBTV Rep linked to a maltose-binding protein exhibited both site-specific nicking and joining activities. These activities were dependent on the presence of Mg2+ or Mn2+, but did not require ATP. The fusion protein specifically cleaved ssDNA between bases +7 and +8 of a conserved nonanucleotide loop sequence which is present in the virion-strand of the stem-loop common region of each BBTV component. During this reaction, the fusion protein became covalently attached to the 5' end of the 3'cleavage product. After the nicking reactions, the fusion protein was also capable of catalysing the joining of two nicked ssDNA fragments in a site-specific manner. Based on these activities, BBTV Rep would appear to be very similar to the Rep proteins of the geminiviruses. PMID- 9225059 TI - Evidence for the presence of a low-level, persistent baculovirus infection of Mamestra brassicae insects. AB - A laboratory culture of Mamestra brassicae insects (MbLC) harbours a latent or occult baculovirus that resembles M. brassicae multiple nucleocapsid nucleopolyhedrovirus (MbMNPV). Although conventional extraction techniques have failed to detect the presence of virus in MbLC, control virus-free insects (MbWS) died of an MbMNPV-like infection after being fed MbLC fat-body cells. This suggested that the MbLC cells harboured infectious MbMNPV, albeit at low levels. We have also demonstrated that fat-body cells from MbLC, but not from MbWS, contain mRNA specific for the polyhedrin gene and transcriptional factors that are capable of activating baculovirus late and very late gene promoters linked to a reporter gene encoding chloramphenicol acetyltransferase. Our data provide indirect evidence that the latent MbMNPV in the MbLC insects is maintained as a persistent infection, with the expression of viral genes at a low level. PMID- 9225060 TI - Promoter analysis of a cysteine-rich Campoletis sonorensis polydnavirus gene. AB - Promoter activity of the Campoletis sonorensis polydnavirus (CsPDV) WHv1.6 gene was analysed by transient transfection assays in insect cell culture using constructs expressing the CAT gene. Deletions of the WHv1.6 gene promoter were used to define promoter regions important for expression. Progressive deletion of the regions upstream of the TATA box reduced the promoter activity, whereas deletions eliminating the TATA box abolished promoter activity. Cis-activating elements were detected up to 1 kb upstream of the WHv1.6 transcription initiation site (TIS). Promoter elements increasing transcription were detected between -444 and -550 bp and between -831 and -1035 bp relative to the TIS. Analysis of the 3' flanking sequences of the WHv1.6 gene indicated that the polyadenylation signals were the only important elements affecting expression in the constructs. Comparison of promoter regions of four cysteine-rich CsPDV genes revealed homologous sequences that may be important for transcriptional regulation of polydnavirus gene expression in parasitized Heliothis virescens larvae. PMID- 9225061 TI - Report on the workshop entitled: "Modeling chemically induced leukemia- implications for benzene risk assessment". PMID- 9225062 TI - A pilot study of alpha-interferon and plicamycin for accelerated phase of chronic myeloid leukemia. AB - Thirteen patients with accelerated phase of chronic myeloid leukemia (CML-AC) were treated with intravenous plicamycin and subcutaneous alpha-interferon. Two patients stabilized, three patients had partial hematologic responses and one patient had a hematologic complete response with a major cytogenetic response. Two patients, progressing on hydroxyurea, did not respond, but demonstrated re sensitization to hydroxyurea after completion of induction therapy and had prolonged return to chronic phase for 30 months and 25 months. Four non responders subsequently received additional chemotherapy and responded. Median survival of all study patients from the development of accelerated phase of CML was 24 months: substantially longer than other reported series (median 6 months). Plicamycin appears to add efficacy to interferon in the stabilization of accelerated phase of CML. PMID- 9225063 TI - Thy-1 expression on blast cells from adult patients with acute myeloid leukemia. AB - Bone marrow and peripheral blood from 28 adult patients with acute myeloid leukemia (AML) were analyzed for the surface expression of the Thy-1 antigen by dual-colour flow cytometry. The Thy-1 antigen was expressed on greater than 5% of cells from seven patients with the proportions of Thy-1 positive cells ranging from 8.1% to 85.0%. The CD34+ Thy-1+ phenotype was present in all seven cases. The expression of Thy-1 on leukemia cells from patients with AML may need to be considered in the development of methods of normal stem cell isolation from these patients. PMID- 9225064 TI - CD34 expression on acute myelocytic leukemia cells. PMID- 9225066 TI - Bryostatin 1: differentiating agent from the depths. PMID- 9225065 TI - Bryostatin 1 (bryo1)-induced monocytic differentiation in THP-1 human leukemia cells is associated with enhanced c-fyn tyrosine kinase and M-CSF receptors. AB - Bryostatin 1 (bryo1), a naturally occurring macrocyclic lactone derived from the marine bryozoan Bugula neritina is a potent protein kinase C (PKC) activator. In this report, we investigated the role of c-fyn protein, a src-related protein tyrosine kinase (PTK), during bryo1-induced monocytic differentiation in a human leukemia cell line, THP-1. Bryo1 treatment for 24 h inhibited the proliferation of THP-1 cells and caused a major fraction of them to become adherent cells with distinct monocyte/macrophage features and enhanced expression of M-CSF receptors (M-CSFR), a hallmark of mature macrophages. The THP-1 cells in control cultures expressed low but detectable levels of c-fyn proteins. Treatment of THP-1 cells with bryo1 resulted in an enhanced expression of c-fyn proteins, but not c-lyn proteins, another member of the src-family of kinases. The bryo1 treatment also enhanced the levels of both c-fyn tyrosine kinase and autophosphorylation activities in THP-1 cells. Using a combined immunoprecipitation and immunoblot analysis, bryo1 was shown to promote an enhanced association between c-fyn kinase and M-CSFR. The inducing activity of bryo1 was associated with PKC activation; treatment of THP-1 cells with bryo1 led to a rapid and transient elevation of total PKC activity in THP-1 cells. These results show that enhanced expression and activation of fyn kinases are critical events associated with monocytic differentiation induced by bryo1 in THP-1 cells. Our findings may be of clinical relevance, as bryo1 has been used in clinical trials of cancer patients. PMID- 9225067 TI - Protein kinase C is not necessary for transforming growth factor beta-induced growth-arrest in leukemia cell lines. AB - Growth and differentiation of blood cell precursors are regulated by cytokines and hormones by mechanisms which are incompletely understood. Protein kinase C (PKC) isozymes are widely regarded as being important in signal transduction pathways. We have shown that one isozyme, PKC beta, is uniquely important in mediating phorbol ester-induced growth-arrest in the HL-60 myeloid cell line. 1,25-dihydroxyvitamin D3 induces differentiation and growth-arrest in many cells. It upregulates the expression of PKC beta, potentiating the action of phorbol ester. We tested the hypotheses that cytokines, which arrest the growth of hematopoietic cells, do so by activating PKC beta, and that differentiation and growth-arrest induced by 1,25-dihydroxyvitamin D3 is caused by upregulation of PKC beta isozyme gene expression. The influence on growth of combinations of five cytokines (TNF alpha, TGF beta 1, gamma-IFN, IL-1, and G-CSF) and 1,25 dihydroxyvitamin D3 on ten human leukemia cell lines (THP-1, HL-60 S, HL-60 PET, U937, K562, Jurkat, MOLT-4, RPM1 8402, KG-1, and KG-1a) was determined. Four cell lines (THP-1, HL-60 S and PET, and U937) exhibited total growth-arrest when incubated with 1,25-dihydroxyvitamin D3 followed by TGF beta 1. The expression by each cell line of mRNA encoding PKC alpha, beta, and delta, both before and after 24 or 48 h of incubation with 1,25-dihydroxyvitamin D3, was determined. Cell lines sensitive to TGF beta 1 each expressed PKC delta endogenously, or expression was up-regulated with 1,25-dihydroxyvitamin D3. U937 cells underexpressed PKC alpha, and HL-60 PET cells underexpressed PKC beta. These data suggested that PKC delta could be responsible for mediating growth-arrest by TGF beta 1. To test this hypothesis directly, we incubated the cells with two bisindolylmaleimide PKC inhibitors during the addition of 1,25-dihydroxyvitamin D3 and TGF beta 1. Surprisingly, the PKC inhibitors did not block the growth arrest induced by 1,25-dihydroxyvitamin D3 and TGF beta 1. This experiment strongly suggests that neither growth-arrest induced by TGF beta 1 nor the potentiation of this growth-arrest by 1,25-dihydroxyvitamin D3 is mediated by a PKC isozyme which is inhibitable by the bisindolymaleimides. PMID- 9225068 TI - Is PKC activation required for leukemia cell differentiation? PMID- 9225069 TI - Morphological changes and apoptosis in bone marrow from patients with myelodysplastic syndromes treated with granulocyte-CSF and erythropoietin. AB - A study of bone marrow morphology and apoptosis was undertaken in 51 patients with myelodysplastic syndromes (MDS) treated with granulocyte colony-stimulating factor (G-CSF) and erythropoietin (EPO). In 19 of these patients (37%), a significant improvement in the hemoglobin level was found after treatment. Apoptosis was measured using a nick-end labeling (TUNEL) technique. Patients with MDS had a significantly higher percentage of labelled (apoptotic) cells in the bone marrow compared to healthy individuals (56.3 +/- 3.8% vs. 16.2 +/- 1.4%, p = 0.0001). Patients with RAS showed a lower percentage of apoptotic cells than patients with RA (68.5 +/- 9% vs. 46.5 +/- 4.8%, p < 0.05), while patients with RAEB did not differ significantly from either RA or RAS. In the patients who responded to treatment, the bone marrow samples displayed significant morphological changes. The percentages of erythroid precursors and myeloblasts were reduced after treatment, and patients who had ring sideroblasts before treatment also showed a reduction in the percentage of these cells. Total erythroid index also decreased in responding patients. The percentage of apoptotic cells decreased significantly in responding patients (58.8 +/- 4.8% before treatment vs. 44.5 +/- 5.5% after treatment, mean reduction 18.3%, p = 0.0003), whereas no significant change was found in non-responding patients. Our results suggest that one important mechanism behind the positive effects of treatment with G-CSF and EPO is a reduction in the degree of ineffective hematopoiesis in MDS. PMID- 9225070 TI - Apoptosis as a parameter of cytokine treatment in myelodysplasia. PMID- 9225071 TI - On the prognostic value of systemic methotrexate clearance in childhood acute lymphocytic leukemia. AB - The prognostic value of systemic methotrexate clearance (ClMTX) during high-dose therapy was evaluated in a cohort of 42 children with acute lymphocytic leukemia (ALL). As part of an extensive chemotherapy protocol, they had received a total of 293 methotrexate (MTX) infusions in the 6-8 g/m2 dose range. At the termination of the study, when they had all been followed up for 3.5 years or more, 26 of these patients were still in continuous complete remission, whereas 16 had suffered relapse. The intrapatient variability in ClMTX during the eight courses was up to six-fold. In 67% of the patients, the maximum level of ClMTX reached at least twice the minimum value. The coefficients of variation for the intra- and interindividual variability in ClMTX were 9-57% and 26-41%, respectively. The cumulative probability of relapse, estimated by the Kaplan Meier procedure, was increased for patients with a high ClMTX during the initial treatment course, but the difference was not significant on a 5% level. There was no significant relationship between high individual median ClMTX and subsequent relapse of ALL. However, ClMTX during the initial infusion, the time-dependent mean for ClMTX, and the individual patient's median ClMTX, were significant predictors for event-free survival in a Cox proportional hazards regression analysis. The present study demonstrates gross pharmacokinetic variability and unpredictable values of ClMTX in subsequent courses after standardized administration of MTX to paediatric patients with ALL. In spite of the association between ClMTX and prognosis shown by some of the analyses, estimates of ClMTX rates may not necessarily be related to disease outcome in a way that can be exploited to the benefit of the individual patient. PMID- 9225072 TI - Does pharmacokinetic variability influence the efficacy of high-dose methotrexate for the treatment of children with acute lymphoblastic leukemia: what can we learn from small studies? PMID- 9225074 TI - Protein kinase inhibitor-induced alterations of drug uptake, cell cycle and surface antigen expression in human multidrug-resistant (Pgp and MRP) promyelocytic leukemia HL-60 cells. AB - Protein kinase inhibitors staurosporine and CGP 41251, a benzoylated derivative of staurosporine with selective PKC inhibitory activity, reversed the decreased rhodamine 123 uptake in HL-60/VCR (with Pgp-mediated drug resistance) but not in HL-60/ADR (MRP-mediated drug resistance) cells. CGP 41251 reversed the decreased rhodamine 123 uptake in HL-60/VCR cells more efficiently (when compared on a equimolar basis) than staurosporine. However, the protein tyrosine kinase inhibitor genistein unexpectedly modulated the decreased rhodamine 123 uptake in Pgp positive (HL-60/VCR) cells, but not in HL-60/ADR (MRP positive) cells. Cell surface phenotype of both HL-60 drug-resistant cell sublines was compared with that of the parental, drug-sensitive HL-60 cells. Both drug-resistant cell lines expressed markedly decreased levels of cell surface HLA class I antigen in comparison with the parental HL-60 cells. A similar decreased cell surface expression of HLA class II/DR on both drug-resistant, as well as of CD59 (protectin) on HL-60/ADR cells was found. Both protein kinase C inhibitors studied (staurosporine and CGP 41251) exhibited variable effects on cell surface antigen (HLA, ICAM-1, CD59) expression, suggesting complex interactions between PKC-dependent and -independent mechanisms in the regulation of surface antigen expression in these cell lines. Staurosporine differed from CGP 41251 in the cell cycle alterations induced in the HL-60 cell lines examined. Staurosporine induced the accumulation of cells in the G2/M phase of the cell cycle and the appearance of pre-G0 (apoptotic) cells in both examined drug-resistant cell lines. Staurosporine induced the appearance of cells with high DNA content in HL-60/ADR, but not in HL-60/VCR cells. PMID- 9225073 TI - Modulation of p53, WAF1/p21 and BCL-2 expression during retinoic acid-induced differentiation of NB4 promyelocytic cells. AB - The NB4 cell line, established from a patient with APL, carries the t(15; 17) and undergoes differentiation along the granulocytic pathway when exposed to retinoic acid (RA). The NB4 cell line was used as a model for exploring the expression of genes and proteins implicated in growth regulation, differentiation and apoptosis during treatment with RA. NB4 cells undergo a series of cytological and molecular alterations during RA treatment--Day 1: cell differentiation marked by an increase in CD11b is evident. Day 2: WAF1/p21 mRNA and then protein rise, though they drop 2 days later. Day 3: the percentage of cells in S phase begins to decrease and G1 arrest begins. Day 4: p53 mRNA level and then protein levels fall. Day 5: CD11b/BCL-2 double staining cells are markedly reduced. No signs of apoptosis were observed after up to 8 days of treatment with RA. These results demonstrate that NB4 cells treated with RA rapidly differentiate and arrest at G1 phase concurrent with p53-independent WAF1/p21 induction; in addition, phenotypic differentiation appears to commence before changes in cell cycle progression. An explanation for the decrease in p53 as well as the lack of apoptosis immediately after BCL-2 downregulation will require further study. PMID- 9225075 TI - The effects of vinblastine on the expression of human immunodeficiency virus type 1 long terminal repeat. AB - Previous work by our group has demonstrated induction of the HIV-LTR following exposure of cells to various DNA-damaging agents such as ultraviolet (UV) light, cisplatin, and doxorubicin. The current experiments were designed to determine the relative effects of the anti-mitotic drug vinblastine on expression of the HIV-LTR. Using human cervical carcinoma (HeLa) cells stably transfected with the chloramphenicol acetyl transferase (CAT) reporter transcriptionally driven by the HIV-LTR promoter, we demonstrated a 9-10-fold induction at 48-72 h following vinblastine treatment. Previous experiments had demonstrated repression of cisplatin or doxorubicin-mediated HIV induction by treatment with salicylic acid. The vinblastine induction also was repressed by salicylic acid treatment, but not by treatment with indomethacin, suggesting a role for the NF kappa B pathway in the inductive response. When UV exposure was coupled to the vinblastine treatment, there was no additive or synergistic effect evident, suggesting similar paths of induction between the two agents. Northern blots demonstrated that these agents were operating at the level of transcription and that salicylic acid inhibited vinblastine-mediated induction of HIV-LTR-CAT mRNA only if administered at the same time as vinblastine; addition of salicylic acid 2 h later had no effect on transcript accumulation. All combinations of treatments with vinblastine and/or salicylic acid markedly reduced cell survival. PMID- 9225076 TI - Achievement of a complete cytogenetic response with hydroxyurea in a patient with chronic myelogenous leukemia. AB - Hydroxyurea rarely produces a complete cytogenetic remission in patients with chronic myelogenous leukemia (CML). In this report, we describe one case of the CML patient who achieved complete cytogenetic remission (no Ph chromosome in 20 25 metaphase cells) by treatment with hydroxyurea alone. By the fluorescent in situ hybridization (FISH) methodology using bcr/abl specific translocation probe, sequential bone marrow specimens from the patient showed the characteristic 9;22 translocation at a higher rate (9-10%) than the normal control range (2.49-4.88%) at the time of complete cytogenetic remission. Thus, it is suggested that FISH is a more sensitive method to detect the bcr/abl fusion gene than conventional cytogenetic analysis for the detection of minimal residual disease in CML. PMID- 9225077 TI - High susceptibility of an Epstein-Barr virus-converted Burkitt's lymphoma cell line to cytotoxic drugs. AB - Susceptibility to cytotoxic drugs was studied using an Epstein-Barr virus (EBV) genome-negative Burkitt's lymphoma (BL) cell line (Ramos) originating from the tumor cells and an in vitro EBV-converted Ramos cell line (B7). Decreased doubling time (DT) with increased saturation density (SD) and elevated [3H]thymidine incorporation were shown in B7, indicating more rapid cell proliferation and growth. However, B7 was highly susceptible to cytotoxic drugs when compared to Ramos. These results indicated whether the presence or absence of EBV genome in tumor cells may be beneficial for evaluation of susceptibility to cytotoxic chemotherapy in patients with BL. PMID- 9225078 TI - Early transformation to acute myeloblastic leukaemia with the acquisition of inv(16) in Ph positive chronic granulocytic leukaemia. PMID- 9225079 TI - Future perspectives in gastroenterology. Proceedings of an international conference. Leipzig, Germany, June 6-8, 1996. PMID- 9225080 TI - The stomach as an endocrine organ. PMID- 9225081 TI - Acid secretion and Helicobacter pylori. PMID- 9225082 TI - Immunopathology of Helicobacter pylori gastritis. PMID- 9225083 TI - Helicobacter pylori infection in ulcer pathogenesis. PMID- 9225084 TI - Cell volume signalling, osmolytes and liver function. PMID- 9225085 TI - Biliary secretion: future perspectives. PMID- 9225086 TI - Newer pathogenetic concepts in cholesterol gallstone formation: a unitary hypothesis. PMID- 9225087 TI - Update hepatitis A-G. PMID- 9225088 TI - Pathogenesis of liver fibrogenesis. PMID- 9225089 TI - Regulation of human pancreatic secretion. PMID- 9225091 TI - Pathogenesis of acute pancreatitis. PMID- 9225090 TI - Stimulus-secretion coupling of pancreatic digestive enzyme secretion. PMID- 9225092 TI - Acute phase reaction of the exocrine pancreas. PMID- 9225093 TI - Regulation of interdigestive gastrointestinal motility and secretion. PMID- 9225094 TI - The intestine, an endocrine organ. PMID- 9225095 TI - Pathogenesis of inflammatory bowel disease. PMID- 9225096 TI - Recent advances in gastric cancer. PMID- 9225097 TI - Molecular genetics of colorectal cancer. PMID- 9225098 TI - Hepatocellular carcinoma. PMID- 9225099 TI - Endoscopic ultrasonography in upper gastrointestinal and pancreatic disease. PMID- 9225100 TI - Immunohistochemistry. PMID- 9225101 TI - Gene level diagnostics in gastroenterology. PMID- 9225102 TI - Use of peptide receptor agonists and antagonists for diagnosis and treatment. PMID- 9225103 TI - Immunosuppressive therapy for chronic inflammatory bowel disease. PMID- 9225104 TI - Extracorporeal shock wave lithotripsy. PMID- 9225105 TI - Future perspectives in gastroenterology: laser. PMID- 9225106 TI - Future perspectives of minimal invasive surgery. PMID- 9225107 TI - Organ-preserving surgery in pancreatic diseases. PMID- 9225108 TI - Commentary on the use of immortalized neuroendocrine cell lines for physiological research. PMID- 9225109 TI - Tissue-specific expression and regulation by 1,25(OH)2D3 of chick protein kinase inhibitor (PKI) mRNA. AB - The heat-stable protein kinase inhibitor (PKI) protein is a specific and potent competitive inhibitor of the catalytic subunit of cAMP-dependent protein kinase (PKA). Previously, it has been shown that vitamin D status affects chick kidney PKI activity: a 5- to 10-fold increase in PKI activity was observed in kidneys of chronically vitamin D-deficient chicks and treatment with 1,25-dihydroxyvitamin D3 (1,25[OH]2D3) in cultured kidney cells resulted in a 95% decrease in PKI activity. The authors have recently cloned the cDNA for chick kidney PKI and have used the coding sequence to study the regulation of PKI mRNA. Northern analysis showed the expression of two PKI messages, which are 2.7 and 3.3 kb in size. These mRNAs are expressed in brain, muscle, testis, and kidney, but not in pancreas, liver, or intestine. PKI mRNA steady-state levels are downregulated by 47% in kidneys from vitamin D-replete chicks as compared to vitamin D-deficient chicks. PKI mRNA levels in brain, muscle, and testis are not affected by vitamin D status. Treatment of primary chick kidney cultures treated with 10(-7) M 1,25(OH)2D3 for 24h resulted in a 20-30% decrease in PKI mRNA. 1,25(OH)2D3 treatment does not affect the stability of PKI mRNA as determined by treatment of cell cultures with actinomycin D. This study shows that 1,25(OH)2D3 directly and tissue-specifically downregulates PKI mRNA in the chick kidney. PMID- 9225110 TI - Effects of constant infusion with insulin-like growth factor-I (IGF-I) to immature female rats on body weight gain, tissue growth and sexual function. Evidence that such treatment does not affect sexual maturation of fertility. AB - Plasma levels for insulin-like growth factor-I (IGF-I) steadily increase in female rats between 20 and 40 d of life, and this increase is intimately related to the wellknown growth spurt occurring at this age. Since specific actions of IGF-I related to sexual function have been described at the ovarian and hypothalamic levels, an endocrine role of rising circulating IGF-I levels during sexual maturation cannot be excluded. Therefore, the impact of adult-type plasma IGF-I levels during the juvenile age, on body weight (BW) gain, growth of several organs, sexual development, and fertility has been evaluated. Female Sprague Dawley rats were infused with rhIGF-I (2 and 4 micrograms/g BW/d, using Alzet minipumps), between 20 and 41 d of life. When infusing 2 micrograms/g BW/d, plasma levels for IGF-I were increased 1.5- to 2-fold over controls at all ages studied. They were further increased with the higher dosage, but only after 35 d of age. Plasma levels for insulin-like growth factor binding protein (IGFBP)-1 to -3 were clearly increased. BW gain was significantly increased, but only with the higher dosage. Tail length was never modified. In contrast, a growth acceleration for spleen, kidneys, adrenals, and ovaries was observed with both dosages. The ovarian weight of treated animals represented approx 140% of control animals with the 4 micrograms/g BW/d dosage. Histology of the enlarged ovaries did not reveal any abnormalities. No meaningful modification of the timing of vaginal opening was observed, and fertility was not comprised by previous rhIGF-I infusion during 20-41 d age period. In summary, early exposure to increased (adult-like) plasma IGF-I levels did not modify BW gain or tail length, but affected the development of spleen, kidneys, adrenals, and ovaries. Exposure to supraphysiological plasma IGF-I levels (> 1200 ng/mL), accelerated BW gain and increased the weight of all organs studied. No signs of precocious sexual maturation were seen and fertility was normal. In conclusion, prematurely increased plasma IGF-I levels affected somatotropic parameters, but not the onset of sexual function. PMID- 9225112 TI - Stress-response proteins in human pituitary adenomas. Expression of heat-shock protein 72 (HSP-72). AB - The presence of heat-shock protein 72 (HSP-72) was investigated by immunohistochemistry (IHC) in a series of 28 surgically removed pituitary adenomas including six somatotroph, two mammosomatotroph, five lactotroph, six corticotroph, four null cell adenomas, and three oncocytomas. Overall, 25 tumors (90%) were positive for HSP-72. One somatotroph, one lactotroph, and one null cell adenomas each contained only sparse, small HSP-72 immunoreactive granules and were regarded as negative. The expression of HSP-72 was commonly uneven differing in degree from cell to cell and among various tumors. In most adenomas, the immunoreactivity was seen as fine granules of moderate density, distributed throughout the cytoplasm. In some cells, the immunoreactivity was strong and diffuse. In one somatotroph, two corticotroph, one null cell, and one oncocytic adenomas, nearly all tumor cells were strongly positive. Adenoma cells, located adjacent to capillaries and small vessels, commonly showed a selective and strong immunoreactivity for HSP-72. The fragments of nontumorous adenohypophysial parenchyma also contained fine immunoreactive cytoplasmic granules accumulating in scattered hormone-producing cells in stellate cells. These results show that HSP-72 is expressed in most pituitary adenomas with a mostly focal and less frequently diffuse pattern of overexpression. PMID- 9225111 TI - Placental progonadotropin-releasing hormone (pro-GnRH) in the rhesus monkey. AB - Gonadotropin-releasing hormone (GnRH) has been shown to play a role in the regulation of human chorionic gonadotropin (hCG) secretion by the human placenta. Molecular studies have demonstrated that human placental trophoblast cells synthesize a progonadotropin-releasing hormone (pro-GnRH) identical to its human hypothalamic counterpart. However, far less is known about nonhuman primates. To determine whether pro-GnRH exists in the rhesus placenta, pro-GnRH mRNA was cloned, sequenced, and shown to be 97.6% homologous to its human placental counterpart. A single base difference (base 1167) in the domain encoding GnRH results in the same amino acid, arginine, in position 8, whereas four base differences (bases 1200, 1253, 1268, 1292) in the domain encoding GnRH-associated peptide (GAP) result in four different amino acids in position 19, 37, 42, and 50. The absence of a basic amino acid in position 50 suggests the rhesus sequence may be cleaved to yield GAP peptides different from the human placenta. Thus, these data justify the use of mammalian GnRH in studies of rhesus placental function, but indicate the need to investigate the roles unique GAP peptides may play in placental/uterine function. PMID- 9225113 TI - Bovine granulocyte chemotactic protein-2 is secreted by the endometrium in response to interferon-tau (IFN-tau). AB - Interferon-tau (IFN-tau) is secreted by the bovine conceptus and may regulate synthesis of uterine endometrial cytokines to provide an environment that is conductive to embryo development and implantation. Interferon-tau stimulates secretion of an 8-kDa uterine protein (P8) in the cow. P8 was purified, digested to yield internal peptides, and partially sequenced to determine identity. Two internal peptides had 100% (13-mer) and 92% (12-mer) amino acid sequence identity with bovine granulocyte chemotactic protein-2 (bGCP-2). Bovine GCP-2 is an alpha chemokine that acts primarily as a potent chemoattractant for granulocyte cells of the immune system. A peptide was synthesized based on a region of bGCP-2 that overlapped with a P8 peptide amino acid sequence, coupled to keyhole limpet hemocyanin, and used to generate high titer polyclonal antiserum in sheep. Western blots revealed that bGCP-2 was not released by endometrium from day 14 nonpregnant cows, but was released in response to 25 nM IFN-tau (p<0.05). Uterine GCP-2 exhibited high affinity to heparin agarose, a characteristic shared by all alpha chemokines. This is the first report describing presence of GCP-2 in the uterine endometrium and regulation by IFN-tau. The regulation of bGCP-2 by IFN tau may have important implications for cytokine networking in the uterus during pregnancy. Also, the regulation of inflammation and angiogenesis by bGCP-2 working together with other cytokines may be integral to establishing early pregnancy and implantation in the cow. PMID- 9225115 TI - Human growth hormone fragment (hGH44-91) produces insulin resistance and hyperinsulinemia but is less potent than 22 kDa hGH in the rat. AB - A 17 kDa fragment of human growth hormone (22 kDa hGH), identified as hGH44-191, has lower binding affinity for growth hormone receptors (GHRs), but has been reported to be more potent in producing glucose intolerance in yellow obese mice. Out aim was to investigate this anomaly by comparing acute development of hyperinsulinemia and insulin resistance ("diabetogenic activity") during hGH44 191 or 22 kDa hGH infusion in normal rats. Fasted awake make rats (350-370 g) were infused via a carotid cannula with saline (CON), 22 kDa hGH (at 0.125 micrograms/min), or hGH44-191 (at 0.64 or 0.32 micrograms/min) for 5.75 h. Over the last 2 h, a euglycemic hyperinsulinemic clamp (insulin infusion rate 0.25 U/kg/h) was performed. After 3.75 h infusion, 22 kDa hGH at 0.125 and hGH44-191 at 0.64 micrograms/min produced basal (preclamp) hyperinsulinemia compared to CON. During the clamp, insulin resistance was consistently produced by 22 kDa hGH at 0.125 and hGH44-191, at 0.64 micrograms/min compared to CON. Using specific radioimmunoassays for 22 kDa hGH and hGH44-191, we determined that under conditions of equivalent diabetogenic activity, molar circulating levels of hGH44 191 were 50-60-fold higher than 22 kDa hGH. It was concluded that whereas 22 kDA hGH and hGH44-191 are both capable of generating acute hyperinsulinemia and insulin resistance in the normal rat, the diabetogenic potency of hGH44-191 is not enhanced compared to 22 kDa hGH, and that diabetogenic potency is in accord with the reported lower binding affinity of hGH44-191 to the GHR. PMID- 9225114 TI - Androgen support of lacrimal gland function. AB - The effects of dihydrotestosterone (DHT) (1 mg/kg) on biochemical parameters related to lacrimal secretion, basal tear flow rate, and pilocarpine-stimulated lacrimal gland fluid secretion, in mature ovariectomized rabbits were studied. The effects of the synthetic estrogen diethylstilbestrol (DES) (100 micrograms/kg), on lacrimal gland biochemical parameters in normal mature female rabbits was also studied. Ovariectomy decreased the total serum levels of testosterone (T) by 88.5% and androstenedione by 35.9%, without changing the levels of dehydroepiandrosterone (DHEA) of its sulfate. Ovariectomy caused a significant regression of the lacrimal glands, decreasing total DNA by 35%, and total protein by 22%. DHT treatment of ovariectomized animals prevented lacrimal gland regression, increasing total gland DNA (31%) and total protein (18%). DHT treatment also increases Na+, K(+)-ATPase activity (29%) and beta-adrenergic receptor binding sites (23%) compared to the ovariectomized group. DHT increased pilocarpine stimulated lacrimal gland fluid secretion (13.26 +/- 1.47 microL/min) compared to the ovariectomized group (7.72 +/- 0.41 microL/min), but DHT treatment paradoxically decreased basal tear flow rate (1.02 +/- 0.04 microL/min) as compared to the ovariectomized rabbits (1.96 +/- 0.12 microL/min). DES decreased the total serum T from 59.33 +/- 10.54 pg/mL to 21.5 +/- 6.06 pg/mL. DES decreased total Na+,K(+)-ATPase by 12% and increased beta-adrenergic receptor binding sites by 83.3%. These results suggest that androgens play a major role in supporting lacrimal gland secretory function. Additionally, they suggest that estrogens may influence certain aspects of lacrimal functions, although it is not clear to what extent those actions are elicited directly or indirectly. PMID- 9225116 TI - Vasoactive intestinal peptide (VIP) mediates the effect of estrogens on the dopaminergic tone in the hypothalamic-pituitary axis of ovariectomized (OVX) rats. AB - The role of vasoactive intestinal peptide (VIP) in the regulation of dopamine (DA) concentration in mediobasal hypothalamus (MBH), posterior and anterior pituitary of ovariectomized (OVX) estrogenized rats was studied using passive immunization against VIP with a specific antiserum (a-VIP). Chronic estradiol administration decreased DA concentration in MBH, and in posterior and anterior pituitary, compared to OVX control rats. DA tissue concentration increased following a-VIP administration to control and estrogenized OVX rats. In vitro study of VIP and a-VIP on DA release from MBH in chronically estrogenized OVX rats showed that estrogens decreased DA evoked-release from MBH;a-VIP increased DA evoked-release from MBH of control OVX and estrogenized rats. VIP decreased DA evoked-release from MBH of OVX rats, but had no effect on estrogenized rats. VIP decreased DA tissue concentration in MBH of OVX control but not of estrogenized rats. It is suggested that VIP decreases DA synthesis and release from hypothalamic neurons in female rats, and that VIP partially mediates the inhibitory effect of long-term estrogen administration on DA release from MBH. PMID- 9225117 TI - Modulation of preoptic regulatory factor-2 (porf-2) mRNAs by castration and hypophysectomy. AB - Neuropeptides are central to the regulation of mammalian gender-dependent development and reproduction. Preoptic regulatory factor-2 is a neuropeptide gene that is known to be expressed in rat brain and testis. In the brain, expression is gender-dependent and age-dependent. Tissue-specific transcripts are found in the preoptic area (POA) of the hypothalamus and in the testis. In order to investigate the effects of reproductive hormone status on expression of porf-2 in the male rat, porf-2 transcripts were studied by Northern blot analysis in intact, hypophysectomized, and castrated rat POA, medial basal hypothalamus (MBH), cerebral cortex (CC), testis, and liver. Castration of hypophysectomy increased levels of the brain-specific 0.84 kb 5' porf-2 transcript in the POA, but did not affect levels of this transcript in the CC. There was a small decrease in the MBH following castration. Hypophysectomy also resulted in a fourfold increase in the 5' 1.1 kb testis-specific transcript. The affected transcripts are localized to the cytoplasm. A nontissue specific 3' transcript was also detected. Interestingly, this 0.6 kb transcript became non-detectable in all tissues examined following hypophysectomy. Porf-2 mRNA was also detected in human hypothalamus, testis, adrenal, placenta, and prostate with unique transcripts in each tissue examined . It has been shown elsewhere that porf-2 is a unique single copy gene in the rat genome. These data demonstrate that expression of the porf-2 gene is differentially regulated at the pretranslational level by intrinsic tissue-specific, as well as extrinsic pituitary and gonadal factors. The selected responses to reproductive hormonal status suggest that porf 2 may play a role in hypothalamic pituitary-gonadal interactions. PMID- 9225118 TI - Expression of the preoptic regulatory factor-1 and -2 genes in rat testis. Developmental and hormonal regulation. AB - Hormone-responsive peptides play a vital role in development and regulation of testicular function. The preoptic regulatory factors, porf-1 and porf-2, were originally discovered in the rat brain, but are also expressed in the rat and human testis. In the brain expression is age-related, hormone-responsive, region- specific, and gender-related, suggesting that porf-1 and porf-2 are involved in gender-specific brain development and function. Tissue-specific porf-1 and porf-2 mRNAs are also found in the testis and hypophysectomy may alter testicular porf-2 expression. It was thus of interest to further examine porf-1 and porf-2 expression in the testis to evaluate their potential as hormone-responsive peptides that regulate testicular development and function. Testicular expression of both porf-1 and -2 was analyzed as a function of maturational stage, aging and hypophysectomy by the solution hybridization/nuclease protection assay, and cellular location determined by in situ hybridization histochemistry. Expression was quantitatively compared in normal male rats at 15, 30 and 60 d (n = 4) and at 2, 6, 12, and 24 mo of age (n = 5). During development porf-1 is expressed at a constant level at 15, 30 and 60 d, then declines significantly with advancing age; levels at 24 mo are only 20% of those seen at 2 mo (p < 0.05). In contrast, porf-2 expression is highest at 15 d of age and steadily declines at 30 and 60 d, plateaus in the mature adult (6 and 12 mo), then exhibits an additional significant decline in the aged 24 mo animals (6 vs 24 mo, p < 0.05). Hypophysectomy of young adult rats at day 42 results in increased testicular expression 12 d later of both porf-1 (p < 0.05) and porf-2(p < 0.005) compared to intact 54-d-old rats (n = 5). In situ hybridization histochemistry confirms that both porf-1 and porf-2 are expressed in the mature testis at 60 d of age. Porf-2 mRNA is localized to immature germ cells including spermatogonia and primary spermatocytes. Porf-1 mRNA is associated with mature sperm and at low levels in the Sertoli cell cytoplasm surrounding spermatocytes. These data suggest that porf-2 is a pituitary hormone-responsive factor in the developing testis and that both porf-1 and porf-2 have cell-type specific functions in the germ cell compartment of the mature testis PMID- 9225119 TI - The role of insulin-like growth factor-I (IGF-I) and estradiol in rabbit corpus luteum progesterone production. AB - To determine whether insulin-like growth factor-I (IGF-I) plays a role in rabbit corpus luteum (CL) physiology the authors examined: IGF-I expression, the effect of IGF-I on progesterone (P) production in vitro, and the interaction of IGF-I with estradiol, the primary luteotropin in the rabbit. Northern blot analysis revealed that IGF-I mRNA is present in the rabbit CL throughout pseudopregnanacy. An intact ovarian in vitro perfusion model and dispersed luteal cell culture were used to determine effects of IGF-I on P production and interactions with estradiol. IGF-I significantly stimulated P production compared to control medium by the isolated, intact perfused rabbit ovary and by dispersed, cultured luteal cells. Estradiol alone did not stimulate P production in vitro. Estradiol did augment IGF-I stimulated P production in the intact perfused ovary and in luteal cell culture. These findings support a role for IGF-I in rabbit CL P production. PMID- 9225120 TI - Cytoplasmic Ca2+ in glucagon-producing pancreatic alpha-cells exposed to carbachol and agents affecting Na+ fluxes. AB - The cytoplasmic Ca2+ concentration ([Ca2+]i) was measured with dual wavelength fluorometry in glucagon-producing mouse pancreatic alpha-cells loaded with the indicator fura-2. Spontaneous rhythmic activity in terms of slow oscillations from a basal level was observed at 3 mM glucose. Like in the insulin-secreting beta-cells the generation of [Ca2+]i oscillations in the alpha-cells was affected by the activity of the Na/K pump. Blocking the pump with ouabain resulted in an initial rise of [Ca2+]i followed by gradual return to the basal level. The oscillations were transformed into sustained elevation of [Ca2+]i by 10 mM L glycine, which is cotransported with Na+. A similar but less pronounced effect was obtained when Na+ was cotransported with 10 mM of the nonmetabolizable amino acid alpha-amino-isobutyric acid. L-glycine induced sustained increase of [Ca2+]i also when the oscillatory activity was suppressed by exposing the alpha-cells to 20 mM glucose in the presence of insulin. The observation that carbachol induces a [Ca2+]i response in isolated alpha-cells calls for reconsideration of current ideas that muscarinic stimulation of glucagon release is an indirect effect mediated by adjacent beta-cells. PMID- 9225121 TI - Tissue-specific expression of inhibin/activin subunit and follistatin mRNAs in mid- to late-gestational age human fetal testis and epididymis. AB - Inhibin/activin subunit (alpha, beta A, and beta B) immunoreactive protein localization patterns and cell type specific inhibin alpha-subunit mRNA expression have been examined in early- to midgestational age human fetal testes. The scarcity of available third trimester human fetal tissue has, however prevented a complete examination throughout the gestational period and the cell specific expression of follistatin and beta A- and beta B-subunit mRNAs are currently unknown at any gestational age. In the present study, this gap is filled and report mRNA expression patterns of inhibin/activin subunits in mid- and late-gestational age (21-33 wk) human fetal testes and testicular duct system. We also report the first examination of follistatin mRNA signals in the human fetal gonad is also resent in both tubular and interstitial cells, and beta B-subunit mRNA is expressed in seminiferous tubules, in mid- and late-gestational age human fetal testes. Inhibin/activin beta A-subunit mRNA was detected in the interstitial cells of remarkably well preserved mid (21 and 22 wk) and late (29 wk) gestational age testes, and is the only activin-system factor mRNA also expressed in tissue of the duct system of the testis (smooth muscle cells of the epididymis). Follistatin mRNA signal was equal to background levels in testicular and duct tissues at all ages examined. These cell specific expression patterns suggest prominent and possibly differential roles for the inhibins and activins, unopposed by gonadal follistatin, in the human fetal male reproductive system. PMID- 9225122 TI - Truncated and full-length glucagon-like peptide-1 (GLP-1) differentially stimulate intestinal somatostatin release. AB - Glucagon-like peptide-1(7-36NH2) is a potent stimulator of insulin secretion, as well as of somatostatin-14 (SS-14) release from the pancreatic and gastric D cells. To investigate the possible effects of this peptide on release of intestinal somatostatin (SS-28 and SS-14, rat intestinal cultures were treated with 10(-12)-10(-6) M GLP-1(7-36NH2), as well as with the structurally related peptides, GLP-1(1-36NH2) and GLP-2. Both forms of GLP-1 stimulated does-dependent increases in intestinal somatostatin; secretion reached 643 +/- 126% of controls (p < 0.001) after treatment with 10(-6) M GLP-1(7-36NH2), and 398 +/-76% of controls (p < 0.001) after 10(-6) M GLP-1(1-36NH2). Thus, GLP-1(7-36NH2) was more effective than GLP-1(1-36NH2) in stimulating secretion of intestinal somatostatin like immunoreactivity (SLI) (p < 0.05). GLP-2 did not affect intestinal somatostatin release. Gel permeation analysis demonstrated that 10(-6) M GLP-1(7 36NH2) stimulated SS-28 by 2.9 +/- 0.4-fold and SS-14 by 9.1 +/- 3.7-fold, whereas GLP-1(1-36NH2) exerted equivalent effects (2.8 +/- 0.9-fold) on both forms of somatostatin. These findings define a novel biological role for GLP-1(7 36NH2) in the regulation of intestinal somatostatin secretion, and demonstrate that GLP-1(1-36NH2) exerts unique biological activities in this system. PMID- 9225124 TI - PKC isoenzyme expression and cellular responses to phorbol ester in JEG-3 choriocarcinoma cells. AB - Protein kinase C (PKc) is a key regulatory enzyme involved in the transduction of extracellular growth signals to the cell nucleus. It occurs in several isoforms, the exact functional roles of which have not been established as yet. The tumor promoting agent 12-O-tetradecanoyl-phorbol acetate (TPA) is the classic activator of PKC and modulates the activity of the activating protein-1 (AP-1) transcription factor complex via this pathway. AP-1, in turn, induces cell proliferation in many tissues. In the present study, the PKC isoenzyme expression pattern in JEG-3 choriocarcinoma cells was analyzed. The results were compared with those obtained in HEC-1B endometrium adenocarcinoma cells, which had previously been characterized in this respect. To gain insight into the possible functional consequences of different PKC expression patterns, cell proliferation rates and AP-1 activity in response to TPA in both cell lines was studied. Western blot analysis of the PKC isoenzyme expression pattern revealed that JEG-3 cells are deficient in the PKC alpha, delta, and epsilon isoforms. These isoenzymes are strongly expressed in HEC-1B cells, with the alpha and delta being constitutively active. As opposed to HEC-1B cells, JEG-3 cells did not show an enhanced proliferation rate in response to TPA. Furthermore, TPA-treated JEG-3 cells did not exhibit any change in cell shape and refractility as observed in HEC-1B cells. AP-1 activity, as determined by a transfected AP-1-luciferase reporter plasmid, was induced 10-fold by TPA in JEG-3 cells, yet only threefold in HEC-1B cells. It is concluded from these data that differential expression of a subset of PKCs, e.g., the alpha, delta, and epsilon isoforms, may serve as an indicator of the proliferative potential in response to growth factors and mitogens. Furthermore, our data indicate that the inducibility of AP-1 activity does not necessarily reflect the proliferative capacity of a given cell type in response to classical tumor promoters such as phorbol ester. PMID- 9225123 TI - The steroidogenic acute regulatory (StAR) protein two years later. An update. PMID- 9225125 TI - Chronic administration of growth hormone (GH) to adult chickens exerts marked effects on circulating concentrations of insulin-like growth factor-I (IGF-I), IGF binding proteins, hepatic GH regulated gene I, and hepatic GH receptor mRNA. AB - In young birds, growth hormone (GH) administration has been found to have only a small or even no effect on circulating concentrations of insulin-like growth factor-I (IGF-I). This is in obvious contrast to the situation in mammals. The present study examines the effect of continuous administration of GH in adult male chickens. Plasma concentrations of IGF-I were markedly elevated (2.5-3.0 fold, p < 0.001) in GH-treated chickens. There were also some transient increases in the circulating levels of IGF binding proteins. Adult chickens showed other manifestations of increased responsiveness to GH, including elevated hepatic expression of GH-regulated gene-I (mRNA) with GH treatment (p < 0.05), and a tendency (p < 0.08) for decreased GH-receptor mRNA. In contrast to the changes in circulating concentrations of GH and IGF-I with GH treatment, no changes in plasma concentrations of thyroid hormones, reproductive hormones, glucose, or nonesterified fatty acids were evident. PMID- 9225126 TI - Estrogen replacement in ovariectomized rats results in physiologically significant levels of circulating progesterone, and co-administration of progesterone markedly reduces the circulating estrogen. AB - Estrogen and progesterone replacement in ovariectomized rats in an often-used experimental system for determination of the specific effects of these hormones. In this study, two different delivery systems and two different dosage levels of estrogen, progesterone or a combination of the two have been used. Estrogen and progesterone in the circulation have been measured in response to each treatment. It is reported that estrogen treatment (237.2 +/- 49.2 pg/mL) results in physiologically significant levels of circulating progesterone (11.1 +/- 1.3 ng/mL). Also, co-administration of progesterone (23.7 +/- 2.0 ng/mL) with estrogen decreases the level of estrogen over that seen with estrogen alone (96.7 +/- 19.2 pg/mL with progesterone vs 237.2 +/- 49.2 pg/mL without progesterone). Thus, contrary to expectations, estrogen replacement therapy is not specific to estrogen and some of the antagonistic effects of progesterone are the result of a decrease in circulating estrogen, and not a specific effect on a target tissue. Whereas the mechanism of these effects has not been determined, obvious artifactitous phenomena have been excluded as being their cause. These results could have a major impact on the interpretation of past and future experiments of this kind. PMID- 9225127 TI - Nonclassical secretory dynamics of LH revealed by hypothalamo-hypophyseal portal sampling of sheep. AB - Continuous withdrawal of hypophyseal portal blood from unrestrained sheep has permitted detailed assessments of the pulsatile secretion of gonadotrophin releasing hormone (GnRH). To determine if this blood can also be used to characterize the sensory dynamics of pituitary hormones, patterns of luteinizing hormone (LH) in the hypophyseal portal blood of ovariectomized ewes was compared with previous patterns of GnRH and peripheral LH. Hypophyseal portal blood and jugular vein blood were collected every 5 min from six ovariectomized ewes over 6 12 h. Hypophyseal portal blood contained GnRH-associated, sharply defined LH pulses that were much larger than in the periphery. Pulses of secreted LH (hypophyseal portal LH less peripheral LH) showed much faster rates of rise and fall than peripheral and followed pulses of GnRH by an average of 1.26 min. In contrast to pulses in jugular blood, secreted LH pulses often reached a relatively unchanging interpulse nadir-plateau and thereby approached closely algorithm-estimated, extrapolated baselines. The interpulse baseline concentrations of secreted LH (99.6 ng/mL) in hypophyseal portal blood were 31 fold higher than those for jugular LH (3.23 ng/mL). These elevated concentrations also exceeded mean jugular peak concentrations (11.1 ng/mL) and, thus, primarily must represent newly secreted LH. The non-Gaussian profiles of this secreted LH were substantially more complex than the inputs predicted from jugular LH measurements by deconvolution. Furthermore, regardless of the analytical approach, estimations of the mass of secreted LH in each pulse did not correlate well with inputs predicted by deconvolution or Kushler-Brown pulsefit analysis of corresponding pulses in jugular blood (r2 ranging 0.40-0.48). Among alternative explanations is the possibility of heterogeneity in concentrations of GnRH in the portal vessels and variable distribution within the hypophysis. In summary, assay of hypophyseal portal blood obtained directly from the pituitary provides a method for direct assessment of secretory responses to hypothalamic peptides, and thereby serves as an unmatched method for studying the dynamics of LH secretion in vivo. With this approach, LH is revealed to be secreted as complex, non Gaussian pulses that are far more sharply defined that those in the periphery, include non-GnRH-dependent, secretory components that cannot be predicted by deconvolution and are followed by periods of relatively constant, basal secretion. PMID- 9225128 TI - Regulation of ovarian follicle differentiation in gonadotrophin-stimulated rats. AB - The aim of the present study was to investigate the regulation of the in vitro DNA synthesis of ovarian cells recovered from prepubertal rats 48 h after administration of pregnant mare's serum gonadotrophin alone (granulosa cells) or followed by human chorionic gonadotrophin (luteal cells). Isolated granulosa cells were cultured in serum-free medium, different stimuli added for periods of 48 h, and 3H-thymidine incorporation was measured. Both follicle-stimulating hormone (FSH) and luteinizing hormone (LH) inhibited 3H-thymidine incorporation by cultured granulosa cells in a dose-dependent manner (FSH: 10, 100, 200 ng/mL = 26, 41, 49% inhibition, respectively; LH: 0.1, 1, 10 ng/mL = 11, 37, 75% inhibition, respectively). On the other hand, estradiol was found to stimulate 3H thymidine incorporation in granulosa cells (Estradiol: 5, 50, 500 ng/mL = 17, 37, 76% stimulation, respectively). In luteal cells, the rate of basal 3H-thymidine incorporation was very low (granulosa cells: 2560 +/- 310; luteal cells: 661 +/- 92 cpm/100,000 cells) and not modified by any stimulus. To determine the possible production of an inhibitory growth factor by the early corpus luteum, 3H thymidine incorporation by granulosa cells was assessed in the presence of 10% conditioned media (CM) recovered from luteal cell cultures. A marked inhibition both in basal and estradiol-stimulated 3H-thymidine incorporation was observed (74 and 76% of inhibition, respectively). Results suggest that an inhibitory growth factor produced by luteal cells after luteinizing gonadotrophin stimulus could be involved in the differentiation of growing follicles to corpus luteum. PMID- 9225129 TI - Serotonin (5-HT) stimulates thyrotropin-releasing hormone (TRH) gene transcription in rat embryonic cardiomyocytes. AB - Thyrotropin-releasing hormone (TRH) and its mRNA have been identified in the rat heart, and TRH can enhance cardiomyocyte contractility in vivo. At present, little is known about cardiac TRH gene transcriptional regulation in the heart. Hormones and neurotransmitters, including thyroid hormone (T3), glucocorticoids, testosterone, and 5-HT initiate effects not only in the cardiovascular system, but also in the regulation of hypothalamic TRH. To clarify the potential roles of these modulators upon the cardiac TRH gene transcription, rat TRH promoter activity was assessed in rat embryonic myocyte cells (H9C2) by transient transfection assays. TRH promoter activity was stimulated significantly by dexamethasone (10(-4) M) and testosterone (10(-5) M), and was inhibited by T3 (10(-7) M). Interestingly, the neurotransmitter 5-HT stimulated TRH promoter activity in H9C2 cells, but not in HTB-11 cells. To further clarify this selective role of 5-HT on TRH promoter transcriptional activity in cardiac cells, 5-HT receptor antagonists and agonists were tested. A selective 5-HT2 receptor antagonist blocked 5-HT stimulation, whereas 5-HT agonist analogs caused augmentative effects when combined with 5-HT. Neither 5-HT nor any antagonists or agonists influenced H9C2 cell growth or morphology. These data suggest that 5-HT is an important transcriptional regulator of the cardiac TRH gene. PMID- 9225130 TI - Inhibition of corticosteroid-binding globulin caused by a severe stressor is apparently mediated by the adrenal but not by glucocorticoid receptors. AB - The effect of stress on serum corticosteroid-binding globulin (CBG) was studied in adult male Sprague-Dawley rats. CBG was measured either by a homologous radioimmunoassay (RIA) or by a binding assay (BA) using 3H-corticosterone. Exposure of adult male rats to a severe stressor such as immobilization (IMO) for 1 h did not alter serum CBG levels, but a significant decrease was found after 6 and especially 24 h IMO. This decrease was not observed after 24 h exposure to a milder treatment such as food and water deprivation. The effect of different periods of exposure to two stressors, IMO or restraint, was also studied. The following results were obtained:serum CBG levels were reduced by IMO, but only by restraint; IMO-induced reduction of CBG levels was always observed 24 h after starting exposure to IMO, independently of the actual period of exposure to the stressor; and IMO-induced inhibition of CBG was proportional to the hours of exposure to the stressor. Although IMO-induced inhibition of CBG was prevented by adrenalectomy, a role for glucocorticoid acting through their classical type II receptors is unclear as far as treatment of rats with the glucocorticoid receptor antagonist RU486 (100 mg/kg) did not prevent the inhibition caused by IMO. The present data clearly indicate that acute exposure to a stressor is able to decrease CBG levels provided that duration of exposure to the stressor and its intensity are high and that the effect is tested at least 6 h after the onset of stress. The effect appears to be mediated by some adrenal factor(s) other than glucocorticoids. PMID- 9225131 TI - GnRH receptors and GnRH endocrine effects on luteoma cells. AB - An ovary implanted into the spleen of an ovariectomized rat develops into a luteinized tumor, growing in response to gonadotrophins. Previously, it was shown that in vivo Buserelin, a gonadotrophin-releasing hormone (GnRH) analog, inhibited tumor growth. To determine if GnRH had a direct effect on tumor cells, the presence of GnRH receptors as well as the endocrine effects of buserelin were studied on tumoral tissue. GnRH receptors were present in luteoma in similar concentrations and dissociation constant (Kd) to control estrous ovaries. In vivo treatment with buserelin did not modify luteoma GnRH receptors. In organ incubations, luteoma secreted significantly higher estradiol and lower progesterone than estrous ovaries; addition of buserelin did not modify steroid secretion. The same difference in basal steroid secretion between luteoma cells and luteal cells superovulated prepubertal ovaries was observed in cell cultures. Although luteinizing-hormone (LH)-stimulated progesterone in both kinds of cells, buserelin significantly inhibited LH-stimulated progesterone only in luteoma cells. These results describe clear differences in basal steroid secretion between tumoral and normal tissue. Furthermore, they show that luteoma possess GnRH receptors similar to those in normal ovarian tissue, and that GnRH analogs have endocrine effects on these cells. Therefore, a direct effect of buserelin on luteoma cells can be postulated. PMID- 9225132 TI - Endochondral bone growth during early pregnancy compared with pseudopregnancy in rats. AB - There are physiological and skeletal changes that occur during pregnancy to accommodate the increased calcium needs of late pregnancy and lactation in the rat. Endochondral bone growth is accelerated during early to midpregnancy, but the endocrine basis of this is not clear. The purpose of this study was to define the role, if any, of placental factors in changes in endochondral growth by comparing changes that occur during pregnancy with pseudopregnancy in the rat. Many hormones change during pseudopregnancy, except placental hormones (e.g., placental lactogens) because a placenta is lacking. Rates of endochondral growth were increased during pregnancy and pseudopregnancy compared to age-matched, unmated controls. There were also increases in body weight in both pregnant and pseudopregnant animals. Since the observed changes occur in both pregnant and pseudopregnant animals, this indicates that endocrine factors other than those secreted by placenta are involved in increased growth during early pregnancy. PMID- 9225133 TI - Modulation of FSH receptor phosphorylation correlates with hormone-induced coupling to the adenylate cyclase system. AB - The authors have recently demonstrated that an inhibitor of protein phosphorylation, staurosporine (SSP), can dramatically enhance follicle stimulating hormone (FSH) stimulated cyclic adenosine monophosphate (cAMP) accumulation in rat granulosa cell line (GFSHR-17) overexpressing about 20-fold FSH receptor than primary granulosa cells. Moreover, incubation with SSP can partially release the cells from FSH-induced desensitization. In this work, it was examined whether coupling of FSH receptor to the adenylate cyclase is correlated with the degree of receptor phosphorylation. Immunoprecipitation of FSH receptor after metabolic labeling of the cells with 32P-orthophosphate revealed that preincubation of the cells with SSP resulted in pronounced reduction in FSH receptor phosphorylation compared to control cells, concomitantly with a dramatic increase in FSH-stimulated cAMP accumulation. In contrast, incubation of the cells with saturating dose of FSH, which leads to uncoupling between the receptor and the adenylate cyclase, resulted in enhanced receptor phosphorylation. Moreover, cells preincubated with FSH could be released from desensitization by further incubation with SSP and a significant reduction in FSH receptor phosphorylation. Immunostaining of the cells with FSH receptor antibody reveal a homogenous distribution of the receptor on the surface of SSP treated cells. Some aggregation of the receptor was evident in control cells that were not treated with SSP. In contrast, massive clustering and capping of the receptor molecules were observed on the surface of FSH-stimulated cells. The current data suggest that phosphorylation-dephosphorylation of the receptor molecules play an important role in the degree of coupling between the receptor and the adenylate cyclase system. Moreover, desensitization to FSH stimulation that is implicated with high degree of receptor phosphorylation may lead to aggregation of the receptor molecules on the cell surface. PMID- 9225135 TI - Virtual Symposium on Osteoporosis. PMID- 9225134 TI - Role of interleukin-1 beta, interleukin-6, and TNF-alpha in intestinal maturation induced by dietary spermine in rats. AB - In the present investigation, the authors aimed to evaluate the role of cytokines in intestinal postnatal maturation induced by dietary polyamines. Neonatal rats were administered either saline (8 mumol) orally. Spermine increased interleukin 1 beta (IL-1 beta), IL-6, and TNF-alpha plasma concentration. The maximum concentrations of IL-1 beta, IL-6, and TNF-alpha were, respectively, observed at 4, 4, and 8 h posttreatment. Intraperitoneal (i.p.) injection of IL-1 beta increased the specific activity of sucrase in whole small intestine, whereas the specific activities of maltase and lactase were significantly enhanced only in the jejunum. IL-6 elicited sucrase and increased maltase specific activity in the whole small intestine, but lactase specific activity was not affected. TNF-alpha had no effect on sucrase and maltase specific activity, but a slight augmentation of lactase specific activity was detected in the jejunum. Spermine and spermidine content in the intestine was increased by i.p. injection of IL-1 beta and IL-6. Corticosterone secretion was elevated by single i.p. injection of IL-1 beta, IL 6, or TNF-alpha. These findings suggest that spermine could induce postnatal intestinal development and corticosterone secretion through a cytokine-dependent mechanism. PMID- 9225136 TI - Nasal calcitonin. AB - Nasal calcitonin is a newly approved treatment for established osteoporosis that increases lumbar spine bone mass, is safe, and well tolerated. Fracture efficacy data is not yet available, although preliminary results are promising. The dose for established osteoporosis is 200 IU. The dose for prevention of postmenopausal osteoporosis has not been established. Nasal calcitonin may be analgesic to bone and may be of benefit in glucocorticoid-induced vertebral osteoporosis. Nasal spray calcitonin may be of benefit to the symptomatic patient with acute vertebral fracture, the complex patient, or the patient with established osteoporosis who is intolerant of bisphosphonates or estrogen. PMID- 9225137 TI - Bisphosphonates in the treatment of osteoporosis. AB - Bisphosphonates are compounds derived from pyrophosphate, a byproduct of cellular cleavage of adenosine triphosphate (ATP), and are resistant to alkaline phosphatase by virtue of replacement of oxygen by carbon. The high affinity of the P-C-P structure for hydroxyapatite accounts for deposition in bone. Modification of the two side chains of carbon alters the potency of the drugs. Of those that have either completed or are undergoing clinical trials, the order of increasing potency for inhibition of bone resorption is etidronate, clodronate, tiludronate, pamidronate, alendronate, residronate and ibandronate (potency range: 1 to 10,000). Less than 5% of bisphosphonates are absorbed and the half life is a few hours. The drugs must be given on an empty stomach because food and beverages interfere with gastrointestinal absorption. Of the absorbed fraction, as much as 60% is taken up by the skeleton and the remainder is excreted unchanged in the urine. Etidronate, tiludronate, residronate, and alendronate are given orally, clodronate intravenously, and pamidronate and ibandronate by either route. At lower concentrations, bisphosphonates inhibit osteoclatic bone resorption, whereas at higher concentrations they may inhibit mineralization and cause osteomalacia. Inhibition of mineralization diminishes with increasing potency. In postmenopausal women, etidronate and alendronate for 3 yr were shown to inhibit bone resorption, increase bone mineral density (BMD) of the lumbar spine and hip, and prevent fractures without producing osteomalacia. Bone formation also is reduced as a consequence of diminished bone resorption but reduction is less than the reduction of bone resorption. In higher doses bisphosphonates may cause upper gastrointestinal disturbances but in recommended doses they generally are well tolerated and have an excellent safety profile. PMID- 9225138 TI - Use of estrogen for prevention and treatment of osteoporosis. PMID- 9225139 TI - DNA extraction from paraffin-embedded tissues using a salting-out procedure: a reliable method for PCR amplification of archival material. AB - Many techniques have been described for the extraction of DNA from paraffin embedded tissues. Numerous efforts have been directed at simplification of these methods for rapid analysis using PCR. One disadvantage to some of the simpler procedures is inefficient PCR amplification, and for more involved ones using phenol/chloroform extraction, reduction in the yield of DNA. In the present study we report the use of a novel salting-out procedure that was utilized to extract DNA from 259 separate microdissection specimens of formalin-fixed, paraffin embedded tissue sections. These sections were derived from 97 patients with tumors of the ampulla of Vater resected between 1965 and 1995 at our institution. The mean DNA yield was 22.75 micrograms (median 13.2 +/- 30.25) and the mean 260/280 absorbance ratio was 1.68 (median 1.70 +/- 0.25). All specimens (259/259) were successfully used to amplify K-ras exon 1 by a nested PCR technique. These results indicate that this DNA extraction method produces good yields of quality DNA, even from specimens several decades old. PMID- 9225140 TI - Apoptosis in dopaminergic neurons of the human substantia nigra during normal aging. AB - Morphological and biochemical alterations have been described in neurons of the aged human brain. However, the cell death process associated with neuronal senescence remains to be elucidated. Apoptosis and autophagic degeneration, two modes of programmed cell death described in embryogenesis and tissue renewal in adult, have been observed in nigral dopaminergic neurons in patients with Parkinson's disease. In the present study, we made the hypothesis that programmed cell death may be also involved in the death of nigral dopaminergic neurons occurring during aging. Cell death types were defined by morphological criteria identified at subcellular level. We thus performed an ultrastructural analysis in order to search for apoptotic and autophagic features in melanized neurons of the substantia nigra in four normal aged subjects. Morphological characteristics of apoptosis, such as contact loss with surrounding tissues, cell shrinkage and chromatin condensation, were found in 2% of the total number of melanized neurons analyzed. Although endoplasmic reticulum appeared normal, mitochrondria were markedly shrunken. Fragments of melanized neurons were found in glial cells. Autophagic degeneration or necrosis were not detected in melanized neurons. Signs of oxidative stress, such as vacuolation of mitochondria, were observed in melanized neurons devoid of apoptotic features. These findings demonstrate that apoptosis is involved in cell death of nigral dopaminergic neurons during normal aging. Since morphological abnormalities found in this study, such as marked mitochondrial shrinkage in apoptotic neurons, were not observed in patients with Parkinson's disease, the mechanisms underlying apoptosis may be different in aging and pathology. PMID- 9225141 TI - p53 mutation and protein alteration in 50 gliomas. Retrospective study by DNA sequencing techniques and immunohistochemistry. AB - Alterations of the p53 protein, which is a 53 kD phosphoprotein and gene product of the p53 gene, has been found to play a major role in the genesis of a variety of human malignancies including tumors of the central nervous system. We investigated 50 tumor specimens from primary central nervous system neoplasms. Tissue samples were screened for mutations by the single-strand conformation polymorphism method and detected mutations were sequenced. All tissue specimens were stained immunohistochemically for p53 protein, which when altered accumulates in the nucleus due to prolonged half-life. Mutations were found in six cases, including one pilocytic astrocytoma World Health Organization (WHO) grade I, two astrocytomas WHO grade II, two anaplastic astrocytomas WHO grade III, and one primitive neuroectodermal tumor (PNET). In terms of relative frequency mutations were found mostly in the group of anaplastic astrocytomas WHO grade III. Interestingly, no mutations were found in the group of investigated glioblastomas. P53 immunopositivity did not correlated with the mutations found, whereas the staining index was significantly higher in the cases with detected mutations than in those without. When p53 alterations is seen as an indicator for different pathogenic pathways in glioma formation, this study gives evidence for a difference between anaplastic astrocytoma and glioblastoma. However, since there was a great overlap in p53 immunopositivity and p53 mutation in tumors of different WHO grades and entities, it seems that p53 will not act as a marker molecule neither for tumor entities nor for tumor malignancy. PMID- 9225142 TI - Effects of short-term treatment with CaCl2 or EDTA on the parathyroid glands in pregnant golden hamsters, with special reference to large vacuolar bodies. AB - The large vacuolar bodies in the parathyroid glands of pregnant golden hamsters after administration of CaCl2 or EDTA were investigated. In the parathyroid glands of the pregnant animals 15 min after administration of CaCl2, the mean serum calcium concentration was significantly high when compared to that of the control animals. In the parathyroid glands of the pregnant animals 15, 30 and 60 min after administration of EDTA, the mean serum calcium concentration was significantly low when compared to that of the control animals. In the parathyroid glands of the pregnant animals 15 min after administration of CaCl2, the percentage area occupied by large vacuolar bodies was significantly increased when compared to that of the control animals. In the parathyroid glands of the pregnant animals 15 min after administration of EDTA, the percentage area occupied by large vacuolar bodies was significantly decreased when compared to that of the control animals. PMID- 9225143 TI - Chronic inhibition of NO synthesis produces myocardial fibrosis and arterial media hyperplasia. AB - Pathophysiological effects of nitric oxide (NO)-deficient hypertension are much better known than are the potential morphological changes. Hearts and main arteries were studied in 15 week old male Wistar rats administered NG-nitro-L arginine methyl ester (L-NAME) for 4 weeks. A does of 40 mg/kg/day increased systolic arterial pressure by 30%, while heart rate decreased by 20%. Heart/body weight ratios were not significantly changed. Total cardiac RNA and DNA content and [14C]leucine incorporation into myocardial protein were, however, increased by 15%, 228% and 97%, respectively. Light microscopy of hearts showed subendocardial areas of necrosis along with different stages of healing. Morphometric evaluation demonstrated significant increase in myocardial fibrosis. Serum lactate dehydrogenase increased by 91%. Proliferation cell nuclear antigen (PCNA) immunohistochemistry indicated positive cells in areas of postischemic repair. Chronic inhibition of NO synthase (NOS) resulted in periarterial fibrosis and hyperplasia of the media in coronary arteries and aorta. RNA and DNA content, and [14C]leucine incorporation into protein of aorta increased by 255%, 95% and 49%, respectively. PCNA staining showed numerous positive nuclei in the media of coronary arteries and the aorta. It is concluded that inhibition of NOS leads to systemic hypertension with focal myocardial fibrosis reflecting reparative responses associated to ischemic injury. This sequence of alterations involves impaired arterial relaxation, and uncontrolled vascular medial proliferation attributed to the absence of smooth muscle cell proliferation inhibition by NO. PMID- 9225144 TI - In vitro effects of estradiol on pituitary GH-immunoreactive cells. AB - In order to determine whether estradiol modulates the proliferation and activity of somatotrophic cells in vitro, a study of GH-immunoreactive cells was carried out in pituitary monolayer cultures obtained from male adult rats treated with 10(-6)M estradiol for 3 hours. Cellular activity was evaluated in a morphometric study of GH-immunoreactive cells. The proliferation rate was determined by double immunostaining for GH and PCNA (proliferating cellular nuclear antigen). The results were compared to those obtained from control dishes. Estradiol was seen to increase the cellular (p < 0.05), nuclear (p < 0.01) and cytoplasmic areas (p < 0.05). Estradiol decreased the percentage of proliferating GH-immunoreactive cells (p < 0.05) and the nuclear density of somatotrophs (p < 0.05) when all cells present in the dishes were considered. However, when only GH-immunoreactive cells were considered, estradiol increased the proliferation rate of these cells (p < 0.05). Overall, our results suggest that, in vitro, estradiol stimulates the cellular activity and proliferation of GH-immunoreactive cells in the rat. PMID- 9225145 TI - Ectasias of the subcapsular sinus in lymph nodes of athymic and euthymic rats: a relation to immunodeficiency. AB - This paper describes a morphologically unusual feature occurring in lymph nodes of some aged euthymic animals but mostly athymic animals. It initially consists of small alveole-like excrescences of the cortical wall of the subcapsular sinus. With dilatation, an excrescence becomes an ectasia which expands into the cortex. Observations suggest that ectasias enlarge under the influence of an increased pressure of the afferent lymph of a node. Such condition conceivably results from a greater lymph formation due to inflammation of the drained tissue site, combined with an impairment to the flow of lymph from the subcapsular sinus into medullary sinuses. A probable relation of ectasia formation to immunodeficiency is discussed. This formation results in the atrophy of the affected lymphoid cell populations of a node which likely contributes to aggravate the deficiency of the immune system. PMID- 9225146 TI - Electron microscopic perspectives of gill pathology induced by 1-naphthyl-N methylcarbamate in the goldfish (Carassius auratus Linnaeus). AB - This experiment has clarified the ultrastructural pathology, by scanning (SEM) and transmission electron microscopy (TEM), induced by 1-naphthyl-N- methylcarbamate (carbaryl) in the gills of juvenile goldfish (Carassius auratus Linneaus). Carbaryl is a low toxicity pesticide commonly used in forestry and agriculture and for controlling aquatic weeds and crustacean predators of shellfish, and has been known to cause gill damage in fish and clams. A variety of cellular changes were observed after exposure of goldfish for 96 h to a sublethal dose of 10 mg carbaryl/l of water. SEM revealed secondary lamellar fusion, distortion, thinning, and mucus release. TEM responses included enlargement of subepithelial lymphatic spaces and mitochondrial disruption and distortion of the lamellar covering epithelium. Pillar cells became detached and chloride cells were vacuolated. Fish were able to withstand these changes in subacute experiments due to redundancy in gill surface area. PMID- 9225147 TI - The value of proliferating cell nuclear antigen (PCNA)/cyclin in the assessment of cell proliferation in glomerulonephritis. AB - Proliferating cell nuclear antigen (PCNA)/cyclin is an acidic nuclear protein increasing from the late G1 to S phases of the cell cycle and whose detection parallels other standard methods for assessing cell proliferation. The aim of this study was to investigate PCNA expression in normal and diseased human kidneys, in order to clarify cell proliferation in renal tissue and to define a possible correlation of its expression with various types of glomerulonephritis (GN). The immunohistochemical avidin-biotin complex (ABC) method was used for the demonstration of PCNA applying the monoclonal antibody PC-10 to paraffin sections from: 10 normal kidneys, 55 renal biopsies with various types of proliferative GN (PGN), 44 renal biopsies with various types of non proliferative GN (NPGN). In PCNA-positive renal biopsies with GN the antigen showed a heterogenous nuclear expression in occasional or few mesangial and glomerular epithelial cells as well as in a greater number of tubular epithelial cells. PCNA was expressed in 20% of normal kidneys and in 38% of renal biopsies with GN. The frequency of PCNA expression was significantly increased in the cases of PGN (47%) compared to that observed in the cases of NPGN (27%) (p = 0.03). PCNA was detected in 10/24 cases of IgA nephropathy, in 3/4 cases of IgM nephropathy, in 5/14 of other types of primary PGN and in 8/13 of secondary PGN. PCNA expression was not correlated with the degree of mesangial cellularity in PGN. Moreover, there was no significant difference in PCNA expression between primary and secondary PGN. PCNA demonstrated an intense expression in the majority of epithelial cells forming cellular crescents in 8/11 cases of PGN. In conclusion, PCNA was observed more frequently in diseased than in normal kidneys. The significant increase in the frequency of PCNA intraglomerular expression in PGN suggests that PCNA has a certain value in the assessment of mesangial proliferation. Moreover, the increased PCNA expression in tubular epithelial cells especially in PGN, indicates their proliferative state and may be correlated with their proposed activation and role in the progression of renal injury. PMID- 9225148 TI - Vitamin E prevents neutrophil accumulation and attenuates tissue damage in ischemic-reperfused human skeletal muscle. AB - Neutrophil accumulation and the consequent production of oxygen-derived free radicals are involved in the pathogenesis of Ischemia-Reperfusion syndrome. In this study we investigated whether a treatment with Vitamin E, which has antioxidant properties, could attenuate the tissue damage by interfering with the influx of neutrophils within the ischemic and reperfused human skeletal muscle. To this purpose, patients undergoing aortic cross-clamping during the surgical repair of aortic abdominal aneurysm were studied as a model of ischemia reperfusion of the lower limb muscles. Muscle biopsies from the right femoral quadriceps of patients not receiving and receiving Vitamin E pretreatment before surgery were taken: a) after the induction of anaesthesia, as control samples, and b) after a period of ischemia followed by 30 min of reperfusion. The tissue samples were either routinely processed for morphological study and immunohistochemical analysis to detect an altered expression of specific endothelial adhesion proteins, such as E-selectin and ICAM-1. The results obtained showed that Vitamin E administration was able to prevent the accumulation of neutrophils within the ischemic and reperfused muscle. This beneficial effect of Vitamin E was due to its ability to hinder the expression of E-selectin and ICAM-1, molecules known to increase the adhesiveness of endothelium to circulating neutrophils. After treatment with Vitamin E a marked attenuation of the reperfusion injury was also evident. In conclusion, Vitamin E treatment may be considered a valuable tool for protection against the ischemia reperfusion damage of human skeletal muscle. PMID- 9225149 TI - Iron binding proteins in gallbladder carcinomas. An immunocytochemical investigation. AB - By immunohistochemistry, the presence of major iron-binding proteins (lactoferrin, transferrin, ferritin) has been investigated in adenocarcinomas (27 cases), adenosquamous carcinoma (1 case), undifferentiated sarcomatoid carcinoma (1 case) and mucinous adenocarcinomas (3 cases) of the gallbladder 10 samples of chronic lithyasic cholecystitis, 4 adenomyomas and 6 tubulo-villous adenomas have also been studied. In a variable share of adenocarcinomas, a positive immunoreactivity for iron-binding antisera was encountered in the cytoplasm, while tubulo-villous adenomas, adenomyomas and the normal epithelium of the gallbladder were generally unreactive. In carcinomatous lesions, the staining intensity was variable between different cases or individual tumour cells. The production of these iron-binding proteins in the gallbladder carcinoma in itself could be related to a greater availability of iron for metabolic processes in the neoplastic cell; alternatively, the cytoplasmic localization of these substances in carcinomatous elements may be a consequence of a defective or impaired function of iron-binding receptors with a modified degree of transmembranous iron transfer. PMID- 9225150 TI - Effects of pneumadin (PNM) on the adrenal glands. 6. Further studies on the inhibitory effect of PNM on dexamethasone-induced atrophy of the rat adrenal cortex. AB - Pneumadin (PNM) is a biologically active decapeptide, which has previously been found to enhanced rat adrenal growth; the mechanism is indirect and probably involves the stimulation of both arginine-vasopressin (AVP) and ACTH release. The effects of 2- and 6-day PNM administration on the atrophic adrenal cortices of rats treated for 8 and 12 days, respectively, with daily subcutaneous injections of 15 or 40 g/100 g body weight of dexamethasone (Dx) were investigated. Morphometry showed that PNM counteracted Dx-induced adrenal atrophy, by preventing the decrease in volume and number of the parenchymal cells. PNM raised aldosterone and corticosterone production of adrenal quarters from Dx-treated rats, but it did not evoke significant changes in the plasma concentrations of the two hormones. The preventive effect of PNM was only partial and almost exclusively evident in rats administered the lower dose of Dx. In light of these findings the following conclusions are drawn: (i) PNM is able to partially overcome the Dx-induced inhibition of the rat hypothalamo-pituitary-adrenal axis, probably by stimulating the pituitary release of AVP and ACTH, that in turn enhance adrenocortical growth; (ii) the PNM-induced improvement of the secretory capacity of atrophic adrenocortical cells is not sufficient to raise the blood level of corticosteroid hormones; and (iii) Dx exerts a direct inhibitory action on adrenocortical cells, which is not counteracted by PNM. PMID- 9225151 TI - The behavior of different types of polytetrafluoroethylene (PTFE) prostheses in the reparative scarring process of abdominal wall defects. AB - Currently one of the most widely used prosthetic materials in the repair of abdominal wall defects, is expanded polytetrafluoroethylene (ePTFE). It has been suggested that its behavior with respect to the reparative process may depend on its structure. The aim of the present study was to evaluate the effect of the structure of 3 ePTFE prostheses on the scarring process in an abdominal-wall defect experimental model. The prostheses employed were the Soft Tissue Patch (STP) which is laminar in structure, Mycro Mesh (MM) which is multilaminar with perforations, and the Dual Mesh (DM) prosthesis which has one non-porous surface. Abdominal wall defects (7 x 5 cm) were created in 36 New Zealand rabbits and repaired using fragments of STP, MM and DM. Follow-up periods were 14, 30, 60 and 90 days post-implant. At these times prostheses were macroscopically examined for the presence of infection and/or rejection and the formation of adhesions to abdominal viscera. Specimens were also taken for microscopic analysis (optical and scanning electron) and for immunohistochemical analysis using the rabbit macrophage-specific monoclonal antibody RAM-11. Labelled macrophage counts were performed at each follow-up session. No cases of infection or rejection were found. Loose adhesions between prosthesis and underlying viscera were observed in 2 of the STP, 4 of the MM and 2 of the DM implants. STP and DM implants were progressively encapsulated by organized connective tissue on both peritoneal and subcutaneous surfaces. Cellular colonization was observed on both STP surfaces and on the porous surface of the DM although no more than a third of the biomaterial was penetrated by cells in either case. Colonization was very slight at prosthesis anchorage points. MM implants differed only in the formation of connective tissue bridges in perforated areas, and cellular infiltration in interlaminar spaces. Macrophage response was similar in the 3 prostheses with a reduction in RAM-11 labelled cells (p < 0.05) between 14 and 90 days post implant. We conclude: a) the 3 types of PTFE prosthesis induced low incidence of adhesion formation between biomaterial and viscera; b) integration mechanism of the 3 prostheses were similar and culminated with the encapsulation of the PTFE by the neoformed tissue; c) the macrophage response induced by the 3 prostheses was similar to that of any reparative process in the absence of biomaterial. PMID- 9225152 TI - Detection of glycoconjugates in the ductus epididymis of the prepubertal and adult horse by lectin histochemistry. AB - This paper describes an approach for studying the structure of glycoconjugates found in the principal cells lining the epididymal duct in adult and prepubertal horses, using ten different lectin horseradish conjugates: Con-A, LCA, WGA, GSA II, SBA, PNA, RCA-I, DBA, UEA-I, and LTA. Saponification and sialidase procedures, followed by lectin binding, were employed to visualize the distribution and to reveal the sequence of sialoglycoconjugates in ductus epididymis. In the adult horse the results demonstrated variations in the content and distribution of glycosidic residues of glycoconjugates in different epididymal regions (caput, corpus, cauda) and vas deferens, suggesting that each epididymal segment has a specific function. In particular, staining of the Golgi zone in the principal cells lining corpus epididymis was interpreted as evidence for synthesis and secretion of glycoconjugates and sialoglycoconjugates. In the prepubertal horse, only the glycocalyx of the epithelial cells lining the epididymal duct showed reactivity toward the different lectins used, suggesting hormonal regulation of the epididymis activity. Additional, the heterogeneity of the lectin staining pattern of the adult horse epididymis reported in this investigation also suggests the existence of different functional segments along the epididymal duct. PMID- 9225153 TI - Postnatal morphologic changes and glial fibrillary acidic protein immunoreactivity in the anteroventral cochlear nucleus of the acoustically deprived gerbil. AB - This study investigated the morphological changes and glial fibrillary acidic protein immunoreactivity (GFAP-IR) in the anteroventral cochlear nucleus (AVCN) of acoustically-deprived gerbils during postnatal development. The mongolian gerbil, Meriones unguiculatus, had been acoustically deprived on the right side or left side by a surgical ligation of the external auditory canal at postnatal day 12-14. No discernible microcysts were located in the ipsilateral AVCN at one, three, six and nine months after monaural ligation. Also, no discernible microcysts were located in the contralateral AVCN at one and three months after monaural ligation. Numerous microcysts were located in the contralateral AVCN at six months after monaural ligation and were slightly reduced in number at nine months after monaural ligation. Some of the microcysts closely apposed to and connected with the blood vessels through a leakage route or channel. A foamy region was found in the superficial granule cell cap of the AVCN. The foamy region became evident in the ipsilateral AVCN at three months after monaural ligation. However, the foamy region became evident in the contralateral AVCN at three and nine months after monaural ligation. Vacuoles were mainly found in the neuronal cells at the junction of the superficial and deep layers in the AVCN. These vacuoles were found in the contralateral AVCN at one, three, six, and nine months after monaural ligation. However, vacuoles were found in the ipsilateral AVCN only at three months after monaural ligation. Morphological changes of the myelin sheath were found to be more severe in the contralateral AVCN than in the ipsilateral. GFAP-IR was located in the superficial layer of the contralateral AVCN at three and nine months after monaural ligation. However, GFAP-IR was found in the superficial and deep layers of the ipsilateral AVCN at three and nine months after monaural ligation. GFAP-IR was also found in the superficial layers of the ipsilateral AVCN at six months after monoaural ligation. Microcysts are presumably derived from the detachment of the myelin sheath from the retracted axons, protrusion of the myelin sheath, and disruption of the myelin sheath. The major conclusions were that (1) microcysts were greatly reduced following acoustical ligation during postnatal development, and (2) blood vessels and GFAP immunoreactive astrocytes may be involved in the depletion of microcysts for maintaining the homeostasis of the microenvironment in the cochlear nuclei. PMID- 9225154 TI - Proliferation and migration kinetics of stem cells in the rat fundic gland. AB - The proliferation and migration of stem cells in the developing and adult rat fundic gland have been studied using BrdU immunohistochemistry and BrdU-GSA II (Griffonia-simplicifolia agglutinin-II) double staining. In the developing rat fundic gland, stem cells were first scattered throughout all levels of the epithelia and then concentrated in the depth of the pits. With the elongation and maturation of the fundic glands, stem cells left the gland base and moved upward. By 4 weeks after birth, the development of the fundic gland was completed and stem cells were confined to a narrow proliferative zone in the isthmus, reaching the adult distribution pattern. In the adult rat fundic gland, stem cells in the isthmus differentiated and migrated upward and downward, replacing the surface mucous cells and glandular cells respectively. For upward migration, it took about one week for stem cells to migrate from the isthmus to the surface. For downward migration, it took about two weeks for stem cells to migrate from the isthmus to the neck, and it took 30-36 weeks to reach the gland unit's blind end. Finally stem cells were lost at the deepest level of the glands. The results obtained by simple topographical distribution in the present experiment agreed well with those obtained by quantitative analysis, suggesting the usefulness of BrdU immunohistochemistry for cell kinetic studies. PMID- 9225155 TI - Expression of argyrophilic nucleolar organizer regions (AgNORs) in the harderian gland of male and female hamsters during postnatal development. AB - The expression of argyrophilic nucleolar organizer regions (AgNORs) was studied in the different secretory cell types of the Harderian gland of male and female Syrian hamsters during postnatal development. Mean AgNOR area was calculated for each cell type in paraffin sections from 7-, 14-, 21-, 28-, 45- and 90-day-old animals. AgNOR content was similar in male type I-cells and in female cells, decreasing in both cell types from the 7th to the 14th day, increasing afterwards at the 21st day, and remaining at relatively stable levels from that point to the end of the study. AgNOR content of male type II-cells was greater than in other cell type studied, and was greater in 45- and 90-day-old animals than in 28-day olds. Changes of AgNOR content in type I-cells of male and female hamsters during the first two weeks seem to be related to changes in proliferative activity while metabolic activity might be responsible for changes taking place later on. Our results also support that male type I- and type II-cells have a different biological behaviour and that type II-cells are far from being degenerating cells. PMID- 9225156 TI - On the mechanism of the transport through Golgi apparatus. AB - The application of an ultrastructural cytochemical method for K(+)-dependent paranitrophenylphosphatase (Na+, K(+)-ATPase) has revealed patterns of plasticity of the Golgi apparatus of neurons in the cerebral cortex of 15-day-old rats. The peripheral part of the cis-most cisterna, being usually reactive, deviates from its regular arrangement in the Golgi stack and contacts with adjacent profile of the granular endoplasmic reticulum or with the next cisterna. The findings prompt the hypothesis that active movements of the cis-cisternae may facilitate and accelerate the transport of nascent proteins to and through the Golgi apparatus. The question about the nature of the contacts between the pointed membrane-bound compartments remains open. PMID- 9225157 TI - Gastric oxyntic cell structure as related to secretory activity. AB - The oxyntic, or parietal cell has two characteristic membrane systems. The mammalian intracellular canaliculi are specialized networks of narrow channels lined with numerous microvilli. The other common to all oxyntic cells is the tubulovesicles, a system of tubules and vesicles. The tubulovesicular compartment is drastically depleted during maximal gastric acid secretion and this is coincident with an increase in the cell surface membrane area. A plausible explanation of this process is the fusion and transfer of tubulovesicular membranes to the plasma membrane. However, for many years there was no convincing evidence of the connections between these two membrane systems. How the tubulovesicular membranes transform into plasma membrane without demonstrable connections has been an enigma to electron microscopists. Recent ultra-high resolution scanning electron microscopic observation on the rat oxyntic cell treated with aldehyde-osmium-aldehyde method revealed that in the resting stage, the tubulovesicles were isolated spherical vesicles. But after tetragastrin stimulation, they were interconnected by slender connecting tubules forming a tubulovesicular network. Then this network was fused to the intracellular canaliculus at relatively few points. These connections between the tubulovesicles and luminal surface membrane was also demonstrated in the frog oxynticopeptic cells. In this review, these membrane transformations as well as changes of the H+/K(+)-ATPase, the lectin binding glycocalyx and the cytoskeleton during secretion will be illustrated and discussed. PMID- 9225158 TI - Unorthodox myogenesis: possible developmental significance and implications for tissue histogenesis and regeneration. AB - During the last few years several reports have described the occurrence of skeletal myogenesis in cells derived from embryonic, fetal and perinatal tissues that usually do not contribute to skeletal muscle in the adult vertebrate body. After a brief description of current ideas on myogenic determination in higher vertebrates, three examples of this unorthodox myogenesis will be described: 1) the occurrence of myogenesis in chick epiblast cells, cultured in isolation in serum-free medium; 2) the presence of cells endowed with myogenic potential in the embryonic mouse neural tube; and 3) the occurrence of spontaneous or induced myogenesis in mesenchymal cells during fetal and postnatal life. A possible embryological basis for unorthodox myogenesis, in relation to gastrulation and morphogenetic fields, is then presented. It is also proposed that unorthodox myogenesis may represent a compensatory mechanism for higher vertebrates that have lost much of the regeneration potential of lower vertebrates. PMID- 9225159 TI - The role of gap junctional intercellular communication (GJIC) disorders in experimental and human carcinogenesis. AB - There is a growing body of evidence supporting the etiologic implication of gap junctional intercellular communication disorders in carcinogenesis. Substantial progress has recently been made both in molecular biology of gap junction and in the field of cancer research. They provide new insights and conceptions of gap junctional disorders in tumor pathology. Modern understanding of the structure, function and regulation of gap junctions, as well as putative mechanisms of its disorders in human and experimental carcinogenesis are discussed in this review with particular emphasis on fast-moving aspects of this problem. PMID- 9225160 TI - Immunocytochemical localization of the vacuolar H(+)-ATPase pump in the kidney. AB - In this article we review immunocytochemical localization studies using a monoclonal antibody raised against the 31 kD subunit of bovine H(+)-ATPase, and indirect immunofluorescent staining. In the proximal tubules there is intense H(+)-ATPase staining along the brush borders of S1 and S2, and linear subvillar invagination staining in S1, S2, and S3 segments. In the thick ascending limb of the loop of Henle there is a mild to moderate degree apical cytoplasmic vesicular staining. In distal convoluted tubule there is mild to moderate degree of H(+) ATPase staining which is sharply delineated along the luminal plasma membrane. In the connecting tubule, the connecting tubule cells show mild to moderate luminal membrane and apical cytoplasmic vesicular staining, and the intercalated cells demonstrate prominent H(+)-ATPase staining which is polarized to either apical or basolateral pole or distributed diffusely throughout the cell. In the cortical collecting duct the principal cells show minimal or no staining while the intercalated cells show very bright H(+)-ATPase staining with 6 identifiable morphologic subtypes based on polarization of the pump to apical or basolateral poles, and the degree of polarization (well polarized or poorly polarized). In the medullary collecting duct the principal cells show no staining and the intercalated cells show prominent H(+)-ATPase staining only in the apical pole. We also describe adaptive responses to different physiologic manipulations e.g., chronic oral acid loads, chronic respiratory acidosis, remnant kidney model, chronic desoxycorticosterone (DOCA) administration, and chronic potassium depletion diet. Moreover, we compare the immunocytochemical localization of the H(+)-ATPase pump of rabbit and kidneys. PMID- 9225161 TI - Hyperactive androgen receptor in prostate cancer: what does it mean for new therapy concepts? AB - Investigations on androgen signaling alterations in the late stages of prostate cancer revealed new molecular mechanisms that may be in part responsible for failure of endocrine therapy. Both primary and metastatic lesions from prostate cancer express androgen receptor protein. Amplification of androgen receptor gene occurs in a subset of prostate cancer patients. Several point mutations of androgen receptor gene have been described; they generate receptors which are functionally activated by androgens, other steroids, and even by antihormones. The frequency of androgen receptor mutations may be high in tumor metastases. Functional activity of androgen receptor is influenced by nonsteroidal factors, such as peptide growth factors and second messengers. Thus, prostate cancer cells adapt to low androgen environment by various mechanisms utilizing androgen receptor. Therefore, new strategies for switching off the androgen receptor are needed. PMID- 9225162 TI - Constitutive and regulated expression of vitronectin. AB - Tissue homeostasis depends on spatially and temporally controlled expression of multifunctional adhesive glycoproteins and their cellular counter receptors, and on a tight regulation of proteolytic enzyme systems. The adhesive glycoprotein vitronectin (Vn) not only regulates adhesive events, but also controls a number of these proteolytic enzyme cascades, including the complement, coagulation, and fibrinolytic systems. However, understanding of the biological functions of this molecule is complicated due to it's conformationally lability and its tendency to self-associate. While plasma Vn is monomeric and lacks exposure of conformationally sensitive epitopes, platelet and tissue-associated Vn are believed to be conformationally altered and multimeric. The latter forms express a functional repertoire distinct from plasma Vn. While little Vn immunoreactivity is detectable in most normal tissues, increased depositions of Vn have been observed in areas of tissue injury and necrosis. Tissue Vn was believed to be plasma-derived, but recent studies indicate that extrahepatic cells have the biosynthetic potential to produce Vn and that its synthesis can be regulated under inflammatory conditions. Here, the constitutive and regulated expression of Vn, its locations in tissues and interaction with other matrix molecules are reviewed and their implications for the functions of this molecule are discussed. PMID- 9225163 TI - Developmental anatomy of the primary olfactory pathway in the opossum Monodelphis domestica. AB - It has been shown in previous studies that the marsupial central nervous system is born at a relatively immature state. Although olfaction is thought to play a role in guiding the locomotion of the newborn, the cellular substrates on which this notion is based have not been systemically investigated. This review article summarises the anatomical development of the primary olfactory pathway in the postnatal Monodelphis. The olfactory epithelium and bulb appear morphologically immature at birth although some of the olfactory neurons are shown to express olfactory marker protein. The olfactory tissues subsequently undergo a rapid sequence of developmental events during the first two postnatal weeks. The evidence shows that the marsupial and eutherian olfactory system share a similar temporal sequence of developmental processes although the former proceeds at a lag time of about 10-14 days compared to that a mice (using the date of birth as a common reference point). Much physiological and behavioral studies remain to be done before we can be certain about the time at which functional maturity is attained in this system. PMID- 9225164 TI - Eosinophils and human cancer. AB - Eosinophils are rare granulocytes that are normally associated with allergic diseases or responses to various parasitic infections. Many types of human cancer, however, are also associated with extensive eosinophilia, either within the tumor itself, or in the peripheral blood, or in both locations. Special techniques such as autoflourescence or immunohistochemistry are sometimes needed to detect the presence of intact and degranulating eosinophils within the tumors. With the help of these techniques, extensive eosinophilia is most often seen in hematologic tumors such as Hodgkin's disease and certain lymphomas; however, many other types of cancer such as colon, cervix, lung, breast, and ovary also contain eosinophilia if diligently sought. Although the presence or absence of eosinophilia within these tumors does not appear to have a major influence on the prognosis of the patient, eosinophils may play an important role in the host interaction with the tumor, perhaps by promoting angiogenesis and connective tissue formation adjacent to the cancer. In addition, tumor-related eosinophilia provides some interesting clues into tumor biology, particularly with regard to production of cytokines by the tumor cells. PMID- 9225166 TI - The hemopoietic system: a phylogenetic approach. AB - Nomadism is a true hemopoietic characteristic during vertebrate phylogeny and ontogeny. This work reviews the mechanism and developmental steps of hemopoiesis, from a phylogenetic point of view. A summary of the principal hemopoietic "foci" along the evolutionary line is also presented. PMID- 9225165 TI - The role of carbohydrate residues in mammalian fertilization. AB - The fertilization process in mammals involves binding and fusion of free-swimming sperm and ovulated eggs. This review focuses on the role of carbohydrate residues in the process of sperm-egg interaction in mammals. The zona pellucida (ZP), the acellular glycoprotein coat surrounding the egg is highly glycosylated and possess both Asn- (N-) linked and Ser/Thr- (O-)linked oligosaccharides, with an extreme structural heterogeneity between the different species. Different carbohydrates on ZP3, such as Galactose in alpha-linkage, N-acetylglucosamine in beta-linkage, were suggested as the complementary sperm receptors, mediating the primary binding between the spermatozoon and the ZP. Several suggested complementary ZP3 binding proteins on the sperm are sp56, beta-1,4 galactosyltransferase and p95. Some carbohydrate residues of the ZP undergo post fertilization modifications that might alter the sperm receptor, thus assisting in the establishment of the block to polyspermy. The studies summarized in this review imply a main role for the carbohydrate residues in the process of sperm egg interaction. PMID- 9225167 TI - Mitosis in the human embryo: the vital role of the sperm centrosome (centriole). AB - The pattern of sperm centrosomal (centriolar) inheritance, centrosomal replication and perpetuation during mitosis of the human embryo is reviewed with a series of electron micrographs. Embryonic cleavage involves repeated mitoses, a convenient sequence to study centriolar behaviour during cell division. After the paternal inheritance of centrioles in the human was reported (Sathananthan et al., 1991a), there has been an upsurge of centrosomal research in mammals, which largely follow the human pattern. The human egg has an inactive non-functional centrosome. The paternal centrosome contains a prominent centriole (proximal) associated with pericentriolar material which is transmitted to the embryo at fertilization and persists during sperm incorporation. Centriolar duplication occurs at the pronuclear stage (interphase) and the centrosome initially organizes a sperm aster when male and female pronuclei breakdown (prometaphase). The astral centrosome containing diplosomes (two typical centrioles) splits and relocates at opposite poles of a bipolar spindle to establish bipolarization, a prerequisite to normal cell division. Single or double centrioles occupy pivotal positions on spindle poles and paternal and maternal chromosomes organize on the equator of a metaphase spindle, at syngamy. Bipolarization occurs in all monospermic and in most dispermic ova. Dispermic embryos occasionally form two sperm asters initially and produce tripolar spindles (tripolarization). Anaphase and telophase follows producing two or three cells respectively, completing the first cell cycle. Descendants of the sperm centriole were found at every stage of perimplantation embryo development and were traced from fertilization through cleavage (first four cell cycles) to the morula and hatching blastocyst stage. Centrioles were associated with nuclei at interphase, when they were often replicating and occupied pivotal positions on spindle poles during mitosis. Sperm remnants were associated with centrioles and were found at most stages of cleavage. Centrioles were found in trophoblast, embryoblast and endoderm cells in hatching blastocysts. Pericentriolar, centrosomal material nucleated astral and spindle microtubules. Abnormal nuclear configurations observed in embryos reflect mitotic aberrations. The bovine embryo closely resembles the human embryo in centriolar behaviour during mitosis. It is concluded that the sperm centrosome is the functional active centrosome in humans and is likely the ancestor of centrioles within centrosomes in foetal and adult somatic cells. The role of the sperm centrosome in embryogenesis and male infertility is discussed, since it is of clinical importance in assisted reproduction. PMID- 9225168 TI - Determinants of axonal regeneration. AB - Axons often regrow to their targets and lost functions may be restored after an injury in the peripheral nervous system. In contrast, axonal regeneration is generally very limited after injuries in the central nervous system, and functional impairment is usually permanent. The regenerative capacity depends on intrinsic neuronal factors as well as the interaction of neurons with other cells. Glial cells may, in different situations, either support or inhibit axonal growth. This review discusses the molecular mechanisms that are involved in promoting and inhibiting axonal regeneration in the nervous system after injuries. PMID- 9225169 TI - Luteinizing hormone on Leydig cell structure and function. AB - The effects of luteinizing hormone (LH) and human chorionic gonadotrophic hormone (hCG) on Leydig cell structure and function are reviewed in this paper under two main headings; responses to LH and hCG stimulation and responses to LH deprivation. With acute LH stimulation, up to 2 hours following the LH injection, there was no change in the volume of a Leydig cell. However, Leydig cell peroxisomal volume and intraperoxisomal SCP2 content showed a rapid and transient change. These changes can be considered to be specific because: i) no other Leydig cell organelle including smooth endoplasmic reticulum (SER) showed such a change, and ii) only the intraperoxisomal SCP2 but not catalase (a marker enzyme for peroxisomes) showed such a change within 30 minutes of LH stimulation. As these changes occurred prior to the peak testosterone levels following this treatment, it is suggested that SCP2 and peroxisomes may have an association with testosterone biosynthesis prior to cholesterol transport into mitochondria. With LH or hCG stimulation for longer periods, i.e. one day or more, the same morphological changes are produced in Leydig cells irrespective of the age of the species, dosage of LH or hCG, and with single or multiple doses. These changes include, Leydig cells hypertrophy and/or hyperplasia, increase in the cellular organelle content (mostly SER and mitochondria) and depletion of lipid droplets. In addition, a recent study showed that Leydig cell peroxisomal volume, SCP2 content, the amount of intraperoxisomal SCP2 and testosterone secretory capacity were also significantly increased in response to chronic LH treatment. The effects of LH deprivation by whatever means (e.g. hypophysectomy, with testosterone and 17 beta-estradiol silastic implants, LH antisera) on Leydig cell structure and function is generally described as opposite to those observed following LH or hCG stimulation. These include Leydig cell hypotrophy and hypoplasia, reductions in the cytoplasmic organelle content in general and specific reductions in SER and peroxisomal volumes, reductions in total catalase and SCP2 in Leydig cells together with reductions in the intraperoxisomal SCP2 content in Leydig cells and their testosterone secretory capacity. PMID- 9225170 TI - Microglia and prion disease: a review. AB - Prion diseases are characterized by the accumulation of PrPSc, an altered isoform of a normal cellular protein, PrPc. The prion hypothesis holds that the process of conformational change from PrPc to PrPSc under the influence of PrPSc constitutes the basic infectious mechanism in prion diseases. It is still unknown whether pathological changes in these diseases, which include spongiform degeneration, nerve cell loss and gliosis, are the result of neurotoxicity of PrPSc, loss of function of PrPc or some other mechanism. Recent in vitro findings using a synthetic peptide of human PrPc implicate microglia as a mediator of pathological changes. The mechanism of the toxicity of this peptide involves activation of microglia oxidative stress, and direct interactions with PrPc synthesizing neurones that reduce their ability to cope with oxidative stress. Microglia thus seem to emerge as a mediator of neuronal degeneration and cell death in prion diseases. PMID- 9225171 TI - The role of dietary fat in obesity. AB - EPIDEMIOLOGY: Epidemiological evidence suggests that a high-fat diet promotes the development of obesity and that there is a direct relationship between the amount of dietary fat and the degree of obesity. The importance of this relationship has been shown in black prepubescent females, who consumed more calories as fat than white females. Moreover, black adult females are heavier and have significant higher cardiovascular disease mortality rates than white females. THE INFLUENCE OF DIETARY FAT ON FOOD INTAKE: An overview of animal studies had indicated that high-fat diets induce greater food intake and weight gain than high-carbohydrate diets. Several factors such as caloric density, satiety properties and post absorbtive processing can contribute to this different response to high-fat diets. Accordingly, the satiating effects after meals with a high fat:carbohydrate ratio is less than for meals with a lower ratio. Some authors have reported that the most important variable influencing meal size is not the level of hunger but the nutrient content of the range of foods consumed. Thus dietary fat has a weak effect on satiety and we suggest that periodic exposure to a high-fat meal, particularly when hunger is high, may be sufficient to lead to overconsumption of energy as fat in obese patients. DIETARY FAT AND FAT BALANCE: Energy balance is well correlated with fat balance in lean controls, whereas there is no correlation with either carbohydrate or protein balances. Several authors have shown that carbohydrate and protein storage is closely regulated by adjusting oxidation to intake, whereas fat is almost exclusively used or stored in response to day-to-day fluctuations in energy balance. The positive relationship between fat intake and lipid oxidation seen in lean controls appears not to be present in obese patients. On high-fat diets, post-obese women failed to increase the ratio of fat to carbohydrate oxidation appropriately. Increasing dietary fat results in preferential fat storage in post-obese women, impaired suppression of carbohydrate and reduction of 24h energy expenditure. CONCLUSIONS: Dietary fat induces overconsumption and weight gain through its low satiety properties and high caloric density. Obese and post-obese subjects do not appear to adapt to dietary fat, and therefore fat storage is increased. PMID- 9225172 TI - Mode of action of orlistat. AB - Gastric and pancreatic lipases are enzymes that play a pivotal role in the digestion of dietary fat. Orlistat, a semisynthetic derivative of lipstatin, is a potent and selective inhibitor of these enzymes, with little or no activity against amylase, trypsin, chymotrypsin and phospholipases. It exerts its effect within the gastrointestinal (GI) tract. Orlistat acts by binding covalently to the serine residue of the active site of gastric and pancreatic lipases. When administered with fat-containing foods, orlistat partially inhibits hydrolysis of triglycerides, thus reducing the subsequent absorption of monoaclglycerides and free fatty acids. This effect can be measured using 24h faecal fat excretion as a representative pharmacodynamic parameter. Orlistat's pharmacological activity is dose-dependent and can be described by a simple Emax model which exhibits an initial steep portion of the dose-response curve with a subsequent plateau (approximately 35% inhibition of dietary fat absorption) for doses above 400 mg/d. At therapeutic doses (120 mg tid with main meals) administered in conjunction with a well balanced, mildly hypocaloric diet, the inhibition of fat absorption (approximately 30% of ingested fat) contributes to an additional caloric deficit of approximately 200 calories. Orlistat does not produce significant disturbances to GI physiological processes (gastric emptying and acidity, gallbladder motility, bile composition and lithogenicity) or to the systemic balance of minerals and electrolytes. Similarly, orlistat does not affect the absorption and pharmacokinetics of drugs with a narrow therapeutic index (phenytoin, warfarin, digoxin) or compounds frequently used by obese patients (oral contraceptives, glyburide, pravastatin, slow-release nifedipine). PMID- 9225173 TI - A one-year trial to assess the value of orlistat in the management of obesity. AB - OBJECTIVES: This paper describes the methodology of a multicentre study designed to assess the efficacy and tolerability of orlistat 120 mg tid as therapy for inducing weight loss in excess of that achieved with a moderately calorie restricted diet alone. The results from a single centre are presented to illustrate the nature of the response. DESIGN: This was a double-blind, randomized, parallel-group, placebo-controlled multicentre study. A four-week, single-blind, placebo run-in period preceded a 52 week double-blind treatment period during which patients received either orlistat or placebo three times a day. At the start of the run-in period, all patients were placed on a diet containing approximately 30% of calories as fat and designed to cause an energy deficit of approximately 600 kcal/d. SUBJECTS: Patients of either sex, more than 18 y of age, with a body mass index (BMI) between 30 and 43 kg/m2 were eligible for enrolment. MEASUREMENTS: Efficacy assessments included: measurements of body weight; anthropometry; quality of life; blood pressure; serum lipids; fasting serum glucose and insulin. Safety assessments included: adverse events; vital signs; ECG; renal and gallbladder ultrasound; haematology; serum biochemistry. OUTCOME: In the single centre there was a reduction in body weight of 5.5 +/- 4.5 (s.d.) kg (5.7% reduction) in the placebo group and 8.6 +/- 5.4kg in the orlistat treated group (8.4% reduction) by six months. Thereafter, the placebo group tended to relapse whereas the orlistat group maintained their loss (2.6% vs 8.4% reduction from initial value at 52 weeks). Total and LDL cholesterol fell by 0.05 mmol/l (1.6%) and 0.14 mmol/l (4.2%), respectively, in orlistat treated patients. The drop-out rate was 48% in the placebo group and 39% in the orlistat group. Intestinal symptoms related to orlistat were significantly increased compared to placebo but were well tolerated. Fat soluble vitamin levels remained within the normal range in the treatment group; the reduction seen in alpha-tocopherol levels in patients receiving orlistat was normalized by the decrease in plasma cholesterol concentrations. Beta-carotene and vitamin D concentrations also decreased in orlistat-treated patients. CONCLUSIONS: This preliminary analysis suggests that orlistat, when used with a health-promoting low-fat and moderately energy-restricted diet, confers advantages in the long-term management of obesity. PMID- 9225174 TI - Results of middle ear ventilation with 'Mangat' T-tubes. AB - Tympanostomy tube placement has been shown to be an effective treatment for recurrent acute otitis media and chronic otitis media with effusion. The Senior author, (K.S. Mangat), considered stiffness and the longer inner limbs of the Goode (Xomed) or Treace (Treace Medical) T-tubes as important factors in the high incidence of complications, and used smaller soft silicone. Mangat-tube (Xomed) with shorter inner limbs. A prospective study was undertaken over a five year period (July 1987-July 1992) which was a continuation of a previous retrospective study of Goode and Treace T-tubes (Mangat, K.S., Morrison, G.A.J., and Ganiwalla, T.M. (1993) Int. J. Pediatr. Otorhinolaryngol. 25, 119-125). 322 Mangat tubes (M tubes) were inserted in 191 patients with persistent otitis media with effusion. The peak ages for insertion were between 4 and 6 years. Spontaneous extrusion occurred in 240 ears (66.5%; 154 patients) at a mean time of 29.3 months. Of these, there were 60 perforations at three months follow-up (18.6%) which fell to 31 perforations after six months (9.6%). Surgical extraction of the M-tube was necessary in 82 ears (22.7%; 50 patients) following persistent otorrhoea or resolution of the condition. Otorrhoea, requiring treatment, was noted in 36 ears (11%). No association was found between the occurrence of infection and the incidence of perforation persisting after a year. There was a higher incidence of persistent perforation in those requiring surgical extraction. The overall persistent perforation rate of only 9.6% would appear to be less than that experienced with Goode or Treace T-tubes. PMID- 9225175 TI - The age of diagnosis of sensorineural hearing impairment in children. AB - OBJECTIVE: To identify factors responsible for delays in diagnosis and treatment of pediatric sensorineural hearing impairment (SNHI), and to assess the thoroughness of medical evaluation in these children. DESIGN: Retrospective analysis. SETTING: State-supported school for the deaf. PATIENTS AND OTHER PARTICIPANTS: 291 children with SNHI, the bast majority of whom are profoundly hearing impaired. Data were collected from the school's database, individual student records, and a parental questionnaire. MAIN OUTCOME MEASURES: (1) The age of diagnosis and treatment of SNHI; (2) actors leading to a delay in diagnosis; (3) current medical evaluations used to determine the etiology of SNHI; and (4) the level of parental satisfaction with the evaluation process. RESULTS: Many children with SNHI experience delays in diagnosis from the time of first suspicion of hearing loss. Children with a risk factor for SNHI are diagnosed no earlier than children without a risk factor. Caucasian children are diagnosed significantly earlier than either Black or Hispanic children, regardless of socioeconomic status. Inconsistent medical evaluation ensues following the diagnosis of SNHI, and parental satisfaction with this process is low. CONCLUSIONS: The average age of diagnosis of SNHI remains unacceptably high. There exists a need to enhance physician awareness of childhood deafness and to develop guidelines for the medical evaluation in cases of pediatric SNHI. Lastly, the importance of parental concern regarding a child's hearing or language development must be re-emphasized. PMID- 9225177 TI - Management of secondary hemorrhage following pediatric adenotonsillectomy. AB - A retrospective study was performed of all patients requiring admission to the Royal Children's Hospital, Melbourne over a 12 year period with secondary haemorrhage following adenotonsillectomy, to determine what percentage of these children received blood transfusions or were returned to the operating room to secure hemostasis, and to identify factors predictive of the need for major intervention. There were 163 children who presented from 2 to 15 days following surgery. Initial management in all cases was establishment of intravenous access, and 151 received intravenous or oral antibiotics. One hundred and forty one were managed without the need for major intervention (87%), including five who had silver nitrate cautery to the tonsillar fossae. Major intervention was required in 22 cases (13%): 5 patients were returned to the operating room for hemostasis; 15 received blood transfusions and 2 underwent both. All surgery was required within 12 h of admissions and all blood transfusions within 24 h. The highest rates of major intervention were in those with fresh bleeding at the time of presentation (38%) and hemoglobin levels less than 100 g/l (36%). For those requiring admission with secondary haemorrhage, a period of observation of 24 h would probably be adequate in the majority of cases to identify those children who will require major intervention by surgery or transfusion. PMID- 9225178 TI - A protocol for otolaryngology-head and neck resident training in pneumatic otoscopy. AB - Otitis media with effusion (OME) is one of the most frequent pediatric diagnoses and is also one of the most common indications for medical or surgical intervention in this age group. Pneumatic otoscopy is the standard for physical diagnosis of a middle ear effusion. We report on our experience with a validation program for otolaryngology-head and neck surgery residents in the use of pneumatic otoscopy to diagnose OME. Four PGY 2 residents sequentially completed a 4 month clinical and didactic training program in pneumatic otoscopy. The trainee sequentially performs pneumatic otoscopy, otomicroscopy, and myringotomy on each patient scheduled for a myringotomy and tube placement the morning of surgery. After each task the trainee is required to state if an effusion is present or not, and the accuracy of the diagnosis is immediately reinforced at the time of myringotomy. The trainee's sensitivity and specificity in diagnosing OME is then calculated for the first and second half of the study period. The trainee is validated in pneumatic otoscopy if the sensitivity is > 80% and > 70% respectively, and the trainee is validated in otomicroscopy if the sensitivity and specificity is > 90% and > 80% respectively. Four residents completed the protocol, and a total of 275 ears were examined. Four residents were validated in pneumatic otoscopy, and three residents were validated in otomicroscopy. We conclude that this protocol allows for accurate documentation of the resident's skill progression and enhances resident education. PMID- 9225176 TI - Perioperative psychosocial interventions for autistic children undergoing ENT surgery. AB - Tonsillectomy/adenoidectomy is one of the most frequently performed operations in the United States. It is therefore likely that pediatric ENG surgeons will encounter autistic and developmentally delayed children in routine practice. Autistic children differ from normal children in that they exhibit severe deficits in language and social functioning; abnormal reaction to stimuli such as light, sound, touch, and pain; attachments to particular unusual objects and rigidly stereotyped routines. They are often mentally retarded. With the increasing importance of managed care and continuous quality improvement, knowledge of how to manage the operative course of such children is crucial for the practising surgeon. Based on research and clinical knowledge of these children, certain psychosocial and medical interventions are presented which may improve the operative course of this population. Using the parent as a consultant; decreasing separation from familiar caretakers, objects, and routines; pre-operative role-playing; tailoring anesthetic induction; and using post-operative distractors are suggested techniques. PMID- 9225179 TI - Otoneurologic evaluation of child vertigo. AB - In this study 282 children with vertigo are subdivided (according to previous experiences) into three large groups: (1) vertigo and cochlear diseases; (2) vertigo as an isolated symptom; and (3) vertigo and C.S.N. diseases. Due to the difficult etiopathogenetic investigation of the patients from the second group, the authors focused on that group as they are less studied, are without associated symptoms (deafness--first group; CNS diseases--second group) and where vertigo appears as an idiopathic and an isolated symptom. A careful anamnestic, clinical and instrumental analysis leads to the following observations: (1) in decreasing order of frequency we find the third group, followed by the first and finally by the second; (2) in spite of the overall lower incidence of the second group, this latter includes the paroxismal benign vertigo (PBV) which is overall the second most frequent vertiginous form (after vertigo due to cranial trauma). In this group the authors underline the reasonably high incidence of the iatrogenic syndromes, insisting on the need of their accurate prevention of these risks; (3) the authors confirm that, nowadays, a reliable etiopathogenetic cause of the apparently isolated vertigo (except for the ascertained iatrogenic forms) cannot be identified. Moreover, in spite of its frequency, PBV is the less known form of vertigo, of which we cannot give a certain diagnosis and which can be identified only the the exclusion of all the other known forms through instrumental and clinical observations. PMID- 9225180 TI - The study of optokinetic 'look' nystagmus in children: our experience. AB - Seventy health children underwent an OKN trial. The authors have chosen to perform only four tests (slow and fast clockwise and counterclockwise OKN) taking into account (in agreement with several international studies) four parameters: sTAP, fTAP, sTSAP, fTSAP (where s and f indicate the velocity of the shifting target slow or fast, TAP is total asymmetry percentage of the SSC--speed of slow components--and TSAP is total asymmetry percentage of saccades). They carried out the statistical analysis of the results, which did not show peculiar difference between child and adult OKN. The result of the test was independent of the side first tested and of sex. The authors have tried to identify the normal range of values more suitable to the study of child OKN; on the basis of the calculation of the 95% percentile the normality range was wider than the range assumed for adults. The authors have also tried to subdivided the results for three different groups of age (I = 3-7 years; II = 8-11 years; III = 12-14 years) in order to observe the degree of OKN maturation with age. From the results obtained the maturation of OKN pathways seems to occur in the 7th year of age for the slow movements; the findings related to the fast movements are more doubtful and need further analysis. Finally, although the number of saccades interposed to the tracings depends on enormous variations unrelated to age, sex and first side tested, our data show their higher incidence during the slow test. PMID- 9225181 TI - Body build--is it a factor in acute subglottic laryngitis? AB - Acute subglottic laryngitis (pseudocroup) is caused by viral infection and usually occurs in children from 6 months to 4 years of age. Obese children are considered to be more susceptible to the disease. In order to evaluate the influence of nutritional status on acute subglottic laryngitis occurrence, an analysis of 193 patients was performed. A group of 70 age-matched healthy children served as the control subjects. The nutritional status of children (body weight and height) was assessed and their percentile positions on the weight and weight-height charts were determined. The recurrence of pseudocroup coexistence of allergy and breast-feeding history were considered in the study. Results of statistical analysis indicate no significant difference in weight and weight height percentile distribution between patients group and controls. The recent changes in child nutrition might be the explanation of decreased susceptibility to pseudocroup among overfed children. PMID- 9225182 TI - Vascular compression of the airway: indications for and results of surgical management. AB - Vascular compression of the airway is a significant cause of respiratory compromise in children. While the indications for surgical repair are sometimes life threatening, they can also be subtle. This retrospective study examines 45 surgical cases of tracheobronchial compromise secondary to vascular compression at a large children's hospital between July 1983 and February 1996. A total of 34 were diagnosed with innominate artery compression, ten with a double aortic arch and one with an anomalous right subclavian artery. The 45 patients, 25 male and 20 female, ranged in age from 12 days to 11 years at surgery (average 13 months). A total of 21 (47%) presented with proven or suspected episodes of cyanosis or apnea. All 45 patients had evidence of vascular compression during microlaryngoscopy and bronchoscopy. The diagnosis was confirmed by magnetic resonance imaging (MRI) in 23/45 (51%), barium swallow in 22/45 (49%) and aortogram in 3/45 (7%). There was one death. One patient had a tracheotomy before surgery and continues to require it after surgery. Complete resolution of symptoms was achieved in 39/45 (87%) with five requiring more than one operation before their symptoms resolved completely. A total of four patients experienced a recurrence of symptoms within a variable length of time after surgery. Surgical indications and treatment alternatives will be discussed. PMID- 9225183 TI - Longitudinal survey of voice quality after pediatric laryngotracheoplasty. AB - This study assesses the effect of pediatric laryngotracheoplasty on voice quality. A group of ten children who underwent laryngotracheoplasty with thyrotomy and anterior cartilage graft were examined two or more times after decannulation and their voices were compared to those of sex- and age-matched controls. Each examination included a laryngoscopy, evaluation by a speech/language pathologist and measurement of maximum phonation times and fundamental frequencies using the Signalyse program. The strength of the voices and the maximum phonation times gradually improved. The fundamental frequency became lower with age, as in the controls. The data suggest that laryngotracheoplasty does not hamper the development of voice with age in children and that the voice improves without any further surgery, although hoarseness remains. PMID- 9225184 TI - Diphtheria or smell: comment on a painting of Goya. PMID- 9225185 TI - Extranasopharyngeal angiofibroma of the inferior turbinate. AB - The first reported case of angiofibroma of an inferior turbinate is presented. The tumour occurred in a 9-year-old boy and was extirpated by subperiosteal dissection of the lateral nasal wall. PMID- 9225186 TI - Infantile schwannoma of the maxillary sinus. AB - A rare case of a 5-year-old female with schwannoma of the maxillary sinus is presented. She had complained of painless swelling of the left cheek and hard palate for a duration of one year. Preoperatively, a CT scan strongly suggested it to be a maxillary cyst with an erupted tooth rather than neoplasm. The tumour was completely removed after embolization of the left internal maxillary artery. The tumour was composed of spindle cells in a palisading pattern and intercellular collagenous fibres. Mitotic figures and atypical nuclei were not observed. Immunohistochemically, the majority of the cells were positive for NSE and S-100 protein, whereas GFA and PCNA showed little immunoreaction. The pathological diagnosis was Antony type A of schwannoma arising in the maxillary sinus. PMID- 9225187 TI - Midline cervical cleft. AB - Midline cervical cleft is a rare congenital anomaly of the ventral neck and its embryological origin has not been clearly established. Less than 100 cases have been reported in the literature. We present a case of midline cervical cleft operated on at the age of 2 months in order to illustrate its clinical presentation and surgical management. PMID- 9225188 TI - A nasopharyngeal dermoid causing neonatal airway obstruction. AB - A case of neonatal respiratory distress due to a pedunculated nasopharyngeal dermoid is presented with its MRI assessment. The dermoid was removed without complications. Nasopharyngeal teratomas are uncommon, consisting of tissues from all three germ layers with varying degrees of differentiation. Symptoms arise during the neonatal period and are associated with airway obstruction. MRI can be performed for contemporary assessment. The first goal of management is to establish a safe and protected airway using complete surgical excision. PMID- 9225190 TI - Pathophysiologic substrate for sustained ventricular tachycardia in coronary artery disease. AB - Sustained ventricular tachycardia (VT) in the presence of coronary artery disease (CAD) is almost always associated with prior infarction. Its mechanism is reentrant excitation and it can be initiated > 95% of the time. Disrupted and delayed endocardial activation and prolonged, fragmented electrograms recorded during sinus rhythm distinguish patients with VT from those with normal ventricles and those of prior infarction without VT. The extent of abnormalities of activation and number of abnormal, fragmented and late electrograms are greatest in patients with sustained VT. These abnormalities are associated with scar tissue separating the viable myocytes. Fragmented electrograms are due to discontinuous activation due to nonuniform anisotropy caused by the scar tissue. Patients with CAD demonstrate depressed excitability and prolonged relative refractory periods (ie, an upward shift in the strength-interval curve) at sites of infarction but effective refractory periods measured at 10 mA comparable to normals and dispersion of refractory periods. However the associated abnormalities of conduction and activation produce an abnormal dispersion of recovery. Intraoperative mapping of patients with CAD has shown that most of the abnormalities of endocardial activation and conduction are in the subendocardial layers and subendocardial resection of these areas cures VT and abolishes delayed, fragmented electrograms and split potentials and normalizes the electrograms recorded from the subjacent tissue. This supports the hypothesis that abnormalities of conduction are the critical pathophysiologic substrate of VT in CAD. PMID- 9225189 TI - Facial nerve neurinoma in a child. AB - Facial nerve neurinoma is a benign tumor, infrequent and exceptional in children. It's clinical manifestation depends on its location and extension, facial palsy being its most frequent sign. Complementary examinations, namely image-diagnosis studies and audiometric tests become essential. Certain diagnosis is made by pathological anatomy. The reported case is a 13-year-old patient suffering from left peripheral facial palsy with an evolution of 8 months who went through a middle cerebral fossa and transmastoid combined tract. PMID- 9225191 TI - Timing of surgical treatment for active native value endocarditis. AB - The aim of the study was to assess the optimum timing of surgical treatment for the active phase of native valve endocarditis. A retrospective study was conducted of the records of patients who had undergone aortic and/or mitral valve replacement for active native valve endocarditis during 1979-94 at Kinki University Hospital. Thirty-three patients with active infective endocarditis of the native valves were treated surgically. Their mean age was 45.4 years (range 11-71). The infective organism was streptococcus in 9 cases, Staphylococcus aureus in 8, and enterococcus in 4 cases. Blood cultures were negative in 9 cases. Of the patients infected with Staphylococcus aureus, 3 died soon after the operation and 1 died later during hospitalization. These 4 patients had been treated medically more than 2 weeks before operation. Another patient who was also treated medically more than 2 weeks before surgery survived. In contrast, all 3 patients infected with Staphylococcus aureus who were operated on within 2 weeks after the onset survived. No early or in-hospital deaths were documented among patients infected with organisms other than Staphylococcus aureus. Among patients who had suffered preoperative embolic episodes, the time from the initial pyrexia to the embolic event was clearly shorter in those infected with Staphylococcus aureus than in those infected with other organisms. Among the former group, 5 out of 6 patients suffered an embolism within 2 weeks of the onset of pyrexia and the remaining 1 within 3 weeks. Thus, in patients presenting with active native valve endocarditis caused by Staphylococcus aureus, surgical treatment should be performed as soon as possible after the onset of pyrexia, preferably within 2 weeks or as soon as the infective organism is identified as Staphylococcus aureus. PMID- 9225192 TI - L-arginine increases exercise-induced vasodilation of the forearm in patients with heart failure. AB - To determine whether L-arginine, a precursor of nitric oxide, can improve exercise-induced vasodilation of the forearm in patients with heart failure, we measured forearm blood flow in 9 patients with heart failure and in 7 age-matched control subjects before and after intra-arterial infusion of L-arginine. Resting forearm blood flow was significantly lower in patients with heart failure than in control subjects (2.34 +/- 0.85 (SD) vs 4.76 +/- 0.77 ml/min per 100 ml, p < 0.001). Endothelium-dependent vasodilation induced by acetylcholine was attenuated in patients with heart failure (p < 0.05). Exercise-induced vasodilation after handgrip exercise was significantly lower in patients with heart failure (p < 0.05). Intra-arterial infusion of L-arginine did not change basal forearm blood flow but significantly augmented acetylcholine-induced vasodilation in both patients with heart failure and control subjects (p < 0.05). Although L-arginine did not affect maximum forearm blood flow after handgrip exercise in control subjects (before, 26.2 +/- 13.5; after, 25.7 +/- 14.3; p = NS), it was increased in patients with heart failure (from 15.2 +/- 4.9 to 24.7 +/- 14.6, p < 0.01). The finding that L-arginine increased both acetylcholine- and exercise-induced vasodilation in patients with heart failure suggests that endothelial dysfunction might play an important role in impaired exercise-induced vasodilation in patients with heart failure. PMID- 9225193 TI - Evaluation of myocardial viability using sequential dual-isotope single photon emission tomography imaging with rest TI-201/stress Tc-99m tetrofosmin in the prediction of wall motion recovery after revascularization. AB - In patients with coronary artery disease (CAD), differentiation between severely ischemic but potentially viable myocardium and irreversibly infarcted tissue is clinically important, particularly when revascularization procedures are considered. Although thallium (TI) cardiac imaging has been shown to be a good tool for investigating myocardial viability in CAD, this tracer shows physical limitations, such as a low photon energy and long half-life. We assessed the results of a rest TI-201/stress Tc-99m tetrofosmin protocol in subjects with prior anterior myocardial infarction. All of the patients had an akinetic or dyskinetic area and more than 75% stenosis in the left anterior descending artery. All of the patients underwent revascularization after the examination. We evaluated the improvement in wall motion after revascularization using the centerline method with contrast left ventricular angiography. Fourteen patients showed reversible defects with the rest TI-201/stress Tc-99m tetrofosmin protocol or in additional TI-201 24 h redistribution images. All 14 patients showed a significant improvement in wall motion after revascularization. Dual-isotope rest TI-201/stress Tc-99m tetrofosmin single photon emission tomography data, acquired separately, may give fast and complete information about myocardial perfusion during stress and at rest, and on about myocardial viability. PMID- 9225194 TI - Clinical application of cardiac output during ramp exercise calculated using the Fick equation--comparison with the 2-stage bicycle ergometer exercise protocol in the supine position. AB - The purpose of this study was to clarify whether the direct Fick method is applicable to the measurement of cardiac output (Q) during ramp exercise. Twelve patients with chronic health failure underwent both a ramp exercise test and a steady-state exercise test. Oxygen intake (VO2), arterial oxygen saturation and mixed venous oxygen saturation were continuously measured using a pulse oximeter and a fiberoptic catheter, and the arteriovenous oxygen difference (a-v O2 diff) and Q were calculated. Both VO2 and a-v O2 diff were significantly lower in the ramp protocol than in the steady-state protocol when they were compared at the same workload. However, the VO2 vs Q relationship and the VO2 vs a-v O2 diff relationship were very similar in the 2 protocols. The difference between Q measured during steady-state exercise and Q calculated at the matched VO2 during ramp exercise was small (3.7 +/- 7.0% and 5.6 +/- 6.6%). The results indicate that, clinically, Q measured by the direct Fick method with simultaneous measurement of VO2 and a-v O2 diff during ramp exercise is a good substitute for the true Q measured by the steady-state protocol. We conclude that Fick Q is applicable to ramp exercise. PMID- 9225195 TI - The effects of class I drugs on the cycle length of sustained ventricular tachycardia and the signal-averaged electrocardiogram. AB - Evaluation of signal-averaged electrocardiograms (SAECGs) has been reported to be useful for predicting the patients with sustained ventricular tachycardia (SVT) combined with organic heart disease and at high risk for sudden cardiac death. However, the relationship between drug efficacy and SAECGs has not been established. We studied the effects of class I drugs on SVT using SAECGs. The study group consisted of 13 patients with SVT who underwent serial electropharmacologic trials. In 7 trials, SVT could not be induced after administration of class I drugs (group 1). In 17 trials, SVT was still inducible (group 2). Filtered QRS in group 2 were significantly prolonged after administration of class I drugs (p < 0.05). The prolongation of the SVT cycle length was correlated with the prolongation of the original QRS (p < 0.05) and with prolongation of the filtered QRS after administration of class I drugs (p < 0.01). The evaluation of the effects of class I drugs on SAECGs may be useful in predicting the inducibility of SVT and SVT cycle length. PMID- 9225196 TI - Increased coronary vasomotor tone in acute myocardial infarction patients with spontaneous coronary recanalization. AB - To clarify the role of coronary spasm or dynamic coronary obstruction in the development of acute myocardial infarction (AMI) with spontaneous recanalization (SR), symptoms in 296 patients with AMI admitted within 24 h after the onset of chest pain were analyzed just before and after onset, and coronary angiograms were analyzed soon after onset. Patients were divided into 3 groups according to the initial angiographic findings in the infarct-related coronary artery (IRCA): group 1 comprised 172 patients with total occlusion (TIMI O); group 2 comprised 57 patients with subtotal occlusion (TIMI 1,2); and group 3 comprised 67 patients with SR (TIMI 3). The incidence of SR was 20.3% at 0-4 h after onset, 22.2% at 4 6 h, 19.7% at 6-12 h, 24.0% at 12-24 h, and 36.0% at 24 h or later. The incidence of SR did not increase significantly as time elapsed. The incidence of angina at rest and variable-threshold angina before the onset of infarction was only 16.2% in group 1, but was significantly higher in groups 2 (64.3%) and 3 (61.9%). The incidence of intermittent chest pain at onset in group 1 (8.4%) was significantly lower than in groups 2 (54.5%) and 3 (38.8%). Vasodilation of the proximal normal segment adjacent to the stenotic site of the IRCA induced by intracoronary nitroglycerin was significantly higher in groups 2 (11.7 +/- 1.2%) and 3 (20.7 +/ 2.6%) than in group 1 (4.0 +/- 0.6%). These results suggest that coronary spasm or dynamic obstruction may be involved in the pathogenesis of thrombus formation or coronary obstruction causing AMI in many Japanese patients. PMID- 9225198 TI - Effects of ischemic preconditioning on ventricular arrhythmias during ischemia and reperfusion using a retrograde blood flow model in dogs. AB - We examined the effects of ischemic preconditioning on ventricular arrhythmias during ischemia and reperfusion from the electrophysiologic point of view by using the retrograde blood flow (RBF) model, which causes severe ischemia. A total of 51 anesthetized dogs were divided into 3 groups. Group 1 (10-min simple occlusion) consisted of 15 dogs; group 2 (10-min RBF) consisted of 20 dogs; and group 3 (10-min RBF with preconditioning) consisted of 16 dogs. Preconditioning consisted of 5 cycles of 2 min of ischemia (RBF) and 5 min of reperfusion. In the subepicardium, myocardial blood flow (MBF) in group 2 was significantly lower than in group 1 or group 3 [group 2 (4.7 +/- 2.3 ml/min per 100 g) vs group 1 (35.0 +/- 5.8) or group 3 (22.0 +/- 4.6); p < 0.01 and p < 0.05 respectively]. However, there were no differences in MBF in the subendocardium between the 3 groups. The incidence of conduction block in the subepicardium was significantly higher in group 2 than in group 1 or group 3 [group 2 (85%) vs group 1 (33%), p < 0.01; vs group 3 (38%), p < 0.01]. There were no differences in the incidence of conduction block in the subendocardium between the 3 groups. During 10-min ischemia, the incidences of ventricular fibrillation (VF) were 7% in group 1, 35% in group 2, and 6% in group 3 (group 2 vs group 1, p < 0.05; and group 2 vs group 3, p < 0.05). During 10-min reperfusion, the incidences of VF were 29% in group 1, 77% in group 2, and 33% in group 3 (group 2 vs group 1, p < 0.05; and group 2 vs group 3, p < 0.05). Ventricular arrhythmias were reduced during both 10-min ischemia and 10-min reperfusion as a result of the improvement in the conduction components by ischemic preconditioning which increased MBF in the subepicardium. PMID- 9225197 TI - Morphological classification of atrial muscle in the atrioventricular junctional area--3-dimensional reconstruction of serial sections of the human heart. AB - To trace anatomical structures that might be associated with the dual atrioventricular (AV) nodal pathway and to investigate the morphologic characteristics of the cells that form these pathways, we examined serial sections of the AV junctional area with a light microscope and reconstructed them 3-dimensionally with a computer. Twelve hearts were obtained at autopsy from patients who had not shown AV conduction disturbances or supraventricular tachycardia before death. The method of Lev et al was used to prepare serial sections. Fascicles of atrial muscle contiguous with the AV node were examined with a light microscope and were classified into 3 groups, on the basis of morphologic characteristics and myocyte diameter. A computer was used to reconstruct 3-dimensionally the course of the fascicles and surrounding structures. At the border of the AV node and bundle of His relatively large myocytes extended directly into the AV bundle from the anterosuperior interatrial septum. Morphologically, the course was considered to be consistent with the fast pathway. In contrast, small cells that entered the AV node from the inferoposterior interatrial septum resembled sinus node cells with few myofibrils and a winding shape. These cells extended from the coronary sinus ostium to the tricuspid valve annulus and are thought to make up the slow pathway. PMID- 9225199 TI - Use of head-up tilt testing to determine a possible cause of unexpected cardiac asystole during epidural anesthesia. AB - Head-up tilt testing is widely used in the diagnosis of syncope of unknown origin. In this report, head-up tilt testing elucidated the etiology of cardiac asystole of unexpected and sudden onset during orthopedic surgery under epidural anesthesia in a 30-year-old woman. Conventional diagnostic approaches were ineffective. Venous pooling in the lower legs as a result of vasodilation and subsequent vagotony due to epidural anesthesia, a condition mimicking orthostatic stress, is proposed as the mechanism of asystole. Follow-up examinations over 16 months revealed no further syncope and a good clinical course. Head-up tilt testing was useful in determining etiology in this case. PMID- 9225200 TI - A case of acute massive pulmonary thromboembolism treated by mechanical clot fragmentation using a percutaneous transluminal angioplasty balloon. AB - Large, bilateral central pulmonary thromboemboli (PTE) led to cor pulmonale and severe hypoxemia in a patient who had undergone Hardy's operation. After several unsuccessful efforts (thrombolysis using a percutaneous catheter and aspiration of the emboli), mechanical clot fragmentation using a percutaneous transluminal angioplasty (PTA) balloon was attempted. This procedure was successful, resulting in a decrease in pulmonary artery pressure from 58/22 (mean 34) mmHg to 20/10 (mean 13) mmHg together with an increase in aortic pressure from 64/36 (mean 45) mmHg to 112/60 (mean 77) mmHg. Thus, mechanical clot fragmentation using a PTA balloon is a promising method for reducing pulmonary artery pressure and increasing aortic pressure in patients with acute PTE. PMID- 9225201 TI - Acute myocardial infarction due to vasospasm in a 13-year-old-boy. AB - We describe an unusual case of acute myocardial infarction due to vasospasm in a 13-year-old boy. He was admitted to our hospital with severe congestive heart failure and shock. He had experienced a feeling of chest oppression with dyspnea while running, which grew worse. He then lost consciousness and was brought by ambulance to our intensive care unit. He had had similar but milder episodes of chest oppression months earlier. The family history revealed that his father had died suddenly from hypertrophic cardiomyopathy and that his grandmother also had hypertrophic cardiomyopathy. On admission, the patient was bathed in a cold sweat, his pulse was weak, and his blood pressure was too low to measure. Coarse crackling and wheezing were audible in both lung fields. Administration of catecholamine and intra-aortic balloon pumping failed to stabilize the hemodynamic variables, but percutaneous cardiopulmonary support proved to be lifesaving. Coronary arteriography performed during his convalescence showed on evidence of atherosclerosis. The acetylcholine provocation test ultimately revealed a diagnosis of acute myocardial infarction due to vasospasm. PMID- 9225202 TI - Thrombolysis and intervention in acute myocardial infarction: an overview. AB - The use of thrombolytic agents in acute myocardial infarction (AMI) has been extensively studied for the past decade and a half and has become the standard of care for most patients presenting early in the course of AMI. Despite this general acceptance, there remains controversy over the choice of thrombolytic, the use of adjunctive anti-platelet and anti-thrombotic agents, the proper role for PTCA, especially direct PTCA, and the potential role for new interventional devices. The intent of this article is to examine in turn each of these areas, reviewing selected data from relevant trials. In so doing we shall develop an overall concept for reperfusion in AMI to quide our ongoing efforts at resolving our remaining therapeutic challenges. PMID- 9225204 TI - Activation of latent TGF-beta at the vascular wall--roles of endothelial cells and mural pericytes or smooth muscle cells. PMID- 9225203 TI - Smooth muscle phenotypes in developing and atherosclerotic human arteries demonstrated by myosin expression. AB - Smooth muscle myosin heavy chains (MHC) exist in multiple isoforms. Rabbit smooth muscle contain at least three types of MHC isoforms; SM1 (204 kDa), SM2 (200 kDa) and SMemb (200 kDa). SM1 and SM2 are specific to smooth muscle, but SMemb is a nonmuscle-type MHC abundantly expressed in the embryonic aorta and in activated mesenchymal cells. We previously reported that these three MHC isoforms are differentially expressed in rabbit during normal vascular development and in experimental arteriosclerosis and demonstrated that MHC isoforms are excellent markers for smooth muscle phenotype. In order to clarify the clinical significance of MHC isoforms, this article will focus on the expression of smooth muscle MHC isoforms in normally developing and atherosclerotic human arteries, especially in coronary arteries. We recently isolated and characterized three cDNA clones encoding human SM1, SM2, and SMemb. The expression of SM2 mRNA in the human fetal aorta was significantly lower as compared to SM1 mRNA but the ratio of SM2- to SM1-mRNA was increased after birth. SMemb mRNA in the aorta was decreased after birth. Immunohistologically, SM1 was constitutively positive from the fetal stage to adulthood in the apparently normal media of the aorta and coronary arteries, whereas SM2 was not detected in fetal arteries of early gestational stage. SM2 was recognized in well-differentiated smooth muscle after perinatal stage. In the human aorta or coronary arteries, unlike in rabbit, SMemb was detected even in the adult. Mild diffuse intimal thickening in the major coronary arteries of the young was found to be composed of smooth muscle cells, reacting equally to three antibodies for MHC isoforms. In thickened but non atheromatous intima, the expression of well-differentiated smooth muscle-specific MHC (SM2) was reduced, especially in the deeper layer. With progression of atherosclerosis, intimal smooth muscle diminished the expression of not only SM2 but also SM1, whereas alpha-smooth muscle actin was well preserved. We conclude from these results that smooth muscle MHC isoforms are important molecular markers for studying human vascular smooth muscle cell differentiation as well as the cellular mechanisms of atherosclerosis. PMID- 9225205 TI - Multi-functional aspects of high density lipoprotein as an anti-atherogenic lipoprotein in vivo: evidence from in vitro experiments using macrophages. PMID- 9225206 TI - Involvement of von Willebrand factor and PGI2 in platelet binding to a partially denuded endothelial monolayer. AB - Platelet binding to an endothelial monolayer was examined after denudation. The binding increased for up to 10 min and thereafter declined gradually. Antibodies against von Willebrand factor (vWF) inhibited the binding. Production of prostacyclin (PGI2) occurred 10 min after endothelial denudation. This study suggests that vWF is involved in the binding during the first 10 min and that PGI2 suppresses the binding thereafter. PMID- 9225207 TI - Effect of modified LDL on the release of NO and PGI2 from rat peritoneal macrophages. AB - Nitric oxide (NO) and prostacyclin (PGI2) have vasodilative and anti proliferative effects on smooth muscle cells (SMC) and an anti-aggregating action on platelets. The present study was designed to elucidate the influence of modified low density lipoprotein (LDL) on the release on NO and PGI2 from rat peritoneal macrophages. Cholesteryl ester (CE) content in macrophages markedly increased on incubation with acetylated LDL (ac-LDL), while NO release did not change. Although incubation with mildly oxidized LDL (m-ox-LDL) and highly oxidized LDL (h-ox-LDL) increased CE content in macrophages, only incubation with h-ox-LDL reduced NO release. PGI2 release from macrophages was not affected by incubation with ac-LDL, m-ox-LDL or h-ox-LDL. These results indicate that the degree of suppression of NO release in macrophages by modified LDL is related to the extent of oxidative modification of LDL itself, but not to the extent of the accumulation of CE in macrophages. Although the role of NO released from macrophages in atherosclerosis is still unclear, the observation of reduced production of NO from macrophages in response to ox-LDL may provide new insight into the role of ox-LDL in the pathogenesis of atherosclerosis. PMID- 9225208 TI - Glycosylation and secretion of lipoprotein lipase by 3T3-L1 adipocytes: effects of brefeldin A. AB - Time courses of synthesis and secretion of lipoprotein lipase (LPL) were examined in 3T3-L1 adipocytes. LPL was glycosylated in the endoplasmic reticulum (ER) within 10 min after synthesis, and was transported after 20-30 min to the trans Golgi where it was converted to the mature form with M(r) = 55,000-58,000, which was resistant to endoglycosidase H (endo H). LPL subunits with M(r) = 55,000 58,000 appeared in the medium within 30 min after synthesis. The effects of brefeldin A (BFA), which inhibits transport of glycoproteins in various types of cells, on secretion and glycosylation of LPL were also examined. BFA completely blocked release of LPL activity into the medium, causing accumulation of the activity in cells. The suppressive effect of BFA on release of LPL activity was reversible. BFA-treated cells synthesized LPL with M(r) = 53,000-55,000 consisting of 2 types of subunits, the main type being totally endo H-sensitive and the other partially endo H-sensitive. No LPL were secreted into the medium by BFA-treated cells. PMID- 9225209 TI - Association of hyperinsulinemia and serum free fatty acids with serum high density lipoprotein-cholesterol. AB - A total of 155 Japanese subjects (79 men and 76 women) who were classified as having normal or borderline glucose tolerance, according to the criteria for the 50-g oral glucose tolerance test (GTT) of the Japanese Diabetes Society, were analyzed for factors related to serum high density lipoprotein (HDL)-cholesterol concentration, especially the responses of insulin and free fatty acid (FFA) after a glucose challenge. In men, significant negative univariate correlations were observed with body mass index (P < 0.01), the summed values of triceps and subscapular skin-folds (P < 0.01), serum insulin concentration at all time intervals, and serum FFA at 30 and 60 min of GTT. Serum insulin at 60, 120, and 180 min, sum insulin, and FFA at 30 and 60 min of GTT were significantly related to serum HDL-cholesterol after adjustment for body mass index and triglyceride concentration. Multiple linear regression analysis with the step-forward method showed that sum insulin (P < 0.01), FFA at 60 min of GTT (P < 0.001), and alcohol consumption (P < 0.01) were independently related to serum HDL-cholesterol concentration. Only the triglyceride concentration was inversely correlated (P < 0.05) with HDL-cholesterol concentration in women. These data indicate that both insulin and FFA concentration, as markers of insulin resistance, apparently influence on HDL kinetics in men, but not in women. The lack of this association in women was appeared to related to the degree of obesity. PMID- 9225211 TI - Hypertriglyceridemia caused by the autoantibody to lipases for plasma lipoproteins: a case report. AB - A 41-year-old female patient with muscle dystrophy, hepatosplenomegaly and tendinous xanthoma showed mild hypertriglyceridemia. The lipoprotein profile in blood showed increases in triglycerides in VLDL and LDL, and a marked decrease of cholesterol in HDL. Chylomicronemia was found, but was not severe. Both lipoprotein lipase and hepatic triglyceride lipase activities were reduced to a level that was only a few percent of the control. Immunoblotting study revealed that the IgG autoantibody in her serum was apparently reactable with hepatic triglyceride lipase and weakly with lipoprotein lipase. Hypertriglyceridemia in this patient is suggested to be due to the autoantibody to these lipases. PMID- 9225210 TI - Effects of an HMG-CoA reductase inhibitor, pravastatin, and bile sequestering resin, cholestyramine, on plasma plant sterol levels in hypercholesterolemic subjects. AB - To study exogenous sterol metabolism during the suppression or stimulation of cholesterol biosynthesis induced by treatments for hyperlipidemia, we determined plasma plant sterol concentrations before and after administration of an HMG-CoA reductase inhibitor, pravastatin, and compared these with changes in these plasma sterol levels by the bile-sequestrating resin, cholestyramine. The effects of the drugs were also studied in a sitosterolemic patient who has had increased plasma levels of plant sterols. Plasma cholesterol levels determined by the HPLC method were decreased significantly after administration of pravastatin. Plasma plant sterol (sitosterol and campesterol) as well as cholestanol concentrations were also significantly reduced. Cholestyramine administration decreased plasma levels of cholesterol, but did not change those of plant sterols in the hypercholesterolemic subjects. Pravastatin had little effect in a sitosterolemic patient on plasma levels of sterols, where cholestyramine decreased the plasma levels of both cholesterol and cholestanol. These results indicate that treatment with the HMG-CoA reductase inhibitor decreases plasma plant sterol concentrations, and suggest that the increased plasma plant sterol levels in sitosterolemia might not be due to the decreased cholesterol biosynthesis in vivo. PMID- 9225212 TI - VLDL receptor in health and disease: interview with a receptor in avian oocytes and mammalian muscle and fat cells. AB - The VLDL receptor is made up of five functional domains that resemble the LDL receptor. In mammals, the receptor is highly expressed in muscle and fat cells, while in chicken, it is abundant in oocytes. The extremely high degree of amino acid conservation of the VLDL receptor during the evolution suggests that the receptor plays an essential role in vertebrates. Recent studies on the chicken VLDL receptor revealed that the receptor plays a key role in the uptake of yolk precursors in oocytes and mediates the growth of oocytes. The VLDL receptor is an essential receptor in avian species and the receptor-deficient mutant hens are sterile and exhibit severe hyperlipidemia with aortic atherosclerosis. PMID- 9225213 TI - Effects of granulocyte-macrophage colony-stimulating factor on the levels of VLDL and LDL receptor mRNAs in vivo. AB - We investigated the mechanism by which granulocyte-macrophage colony-stimulating factor (GM-CSF) lowers plasma cholesterol levels. Recombinant human GM-CSF (rhGM CSF) was administered to normal and Watanabe heritable hyperlipidemic (WHHL) rabbits. Treatment with rhGM-CSF reduced the levels of cholesterol and triglyceride in these animals. In vitro colony assay for hematopoietic progenitors indicated that rhGM-CSF was capable of supporting granulocyte macrophage colony formation in rabbits, suggesting that rhGM-CSF stimulates macrophage function even in rabbits. Northern blot analysis of rabbit very-low density lipoprotein (VLDL) receptor showed that rhGM-CSF elevated the levels of VLDL receptor mRNA 2.6- and 1.8-fold in muscles of normal WHHL rabbits, respectively, 1.5 hours after a single injection. Increases of 1.5- and 1.4-fold were observed in muscles of these rabbits after 5 days of administration. No changes were found in the LDL receptor mRNA levels in liver, spleen or bone marrow. These findings show that the lowering of lipids by GM-CSF may be mediated through the up-regulation of the VLDL receptor mRNA and the enhancement of macrophage function. PMID- 9225215 TI - Lipase activities in post-heparin plasma and tissues, and susceptibilities of lipoproteins in experimental diabetic rats. AB - Heparin administration to diabetic rats caused no change in VLDL, an increase in IDL and a decrease in LDL on electrophoretic analysis of plasma lipoproteins, while the administration to control rats markedly decreased VLDL and increased IDL and LDL. Both hepatic triglyceride lipase (HTGL) and lipoprotein lipase (LPL) activities in the postheparin plasma were lower in the diabetic rats than in the controls, and the reduction of HTGL activity was greater than that of LPL activity in the diabetic rats. The LPL activity in the adipose tissue was lower in the diabetic rats than in the controls, but the activities in the cardiac and skeletal muscles were similar in the two rats. The HTGL-catalyzed fatty acid (FA) releases from the diabetic VLDL and IDL were lower than those from the normal rat VLDL and IDL, while the LPL-catalyzed FA release in the diabetic rats was not different from those in the controls. The decreases in LPL and HTGL activities and the markedly impaired susceptibility of IDL to HTGL coincide well with the postheparin changes in plasma lipoproteins in diabetic rats, an increase in IDL and a decrease in LDL. PMID- 9225214 TI - Endocytic uptake of lysophosphatidylcholine mediated by macrophage scavenger receptor plays a major role in oxidized low density lipoprotein-induced macrophage growth. PMID- 9225216 TI - Intraindividual variations in lipoprotein (a) levels and factors related to these changes. AB - Lp(a) levels are genetically determined and remain stable without major changes throughout lives. However, when an individual's Lp(a) levels are observed over a one-year period, they show spontaneous variation. The rate of intraindividual variation in Lp(a) was observed in 16 patients with hypertension, hyperlipidemia and/or glucose intolerance in a chronic stable state who regularly visited the hospital clinic once a month, at least 10 times during the year, and in whom a total of 42 blood and clinical chemistry tests including serum lipids, Lp(a) and apoproteins were performed. The rate of annual intraindividual variation of Lp(a) averaged out as 16.6%. The rate was 18.8% for isoform S4 (n = 10), 18.6% for S3 (n = 3), and although small in number of subjects, other isoforms showed minor variation rates. There was a significant negative correlation between the rate of variation (y%) and LP(a) level (xmg/dl) r = -0.605, p < 0.05, y = -0.461 x +29.8). Therefore, when Lp(a) was high, the rate of variation (SD%) was low. This was consistent with the finding that the rates of variation were low for isoforms S2, S3S4 and F, whose molecular weights were low, accompanied by high Lp(a) levels. On the other hand, when the relationship between Lp(a) level and the amount of variation (SD mg/dl) was examined, there was no correlation between the two, since the amounts of variation were almost constant at a level of 3.8 mg/dl, regardless of Lp(a) level. The annual variation of Lp(a) level was found to be related to three groups of factors based on comparison of the variations among WHO phenotypes of hyperlipidemias, univariate correlation analysis with the clinical parameters tested, and multivariate analysis: the first group of factors was related to structure and metabolism of very low-density lipoprotein such as triglycerides, phospholipids, apo C-II, C-III, E, A-II and uric acid; the second group was related to thrombosis centering on platelets; and the third group involved those in the acute phase reactions represented by 1 hr and 2 hr erythrocyte sedimentation rates. PMID- 9225217 TI - The long-term effect of eicosapentaenoic acid on serum levels of lipoprotein (a) and lipids in patients with vascular disease. AB - The effects of eicosapentaenoic acid (EPA) on serum lipoprotein (a) (Lp(a)) and other lipid levels in patients with vascular disease were examined. The serum levels of Lp(a), total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL) and very-low-density lipoprotein (VLDL) were measured in 24 patients with vascular disease. An elevated serum Lp(a) level (39 +/- 22 mg/dl) was noted in 9 patients, elevated total cholesterol level (263 +/- 31 mg/dl) in 12 patients, elevated triglyceride level (240 +/- 98 mg/dl) in 10 patients and elevated LDL level (651 +/- 88 mg/dl) in 6 patients before administration of EPA. EPA (1,800 mg/day) was given to these patients for long periods ranging from 6 to 24 months. The serum levels of Lp(a), TC, TG and LDL were lowered significantly (p < 0.05) after EPA administration for 12 and 18 months, for 6, 12, 18 and 24 months, for 18 months and for 12 and 18 months, respectively. These findings indicated that long-term administration of EPA may lower Lp(a) and serum lipids, which is beneficial for patients with various arterial diseases in terms of preventing progression of the disease. PMID- 9225218 TI - In vivo evaluation of DX-9065a, a synthetic factor Xa inhibitor, in experimental vein graft. AB - An in vivo effect of a novel synthetic Xa inhibitor, DX-9065a, was evaluated in a highly thrombogenic venous graft model. A woven Tetron tube graft was interposed in the inferior vena cava of rabbits. All the grafts were completely occluded within 5 hours after a bolus injection of heparin (50 U/kg) given just prior to the grafting. The following agents were continuously given to the respective group of rabbits for 2 h after the bolus injection of heparin; heparin (50 U/kg/h, UFH-group), DX-9065a (0.05 mg/kg/h, DX-group) and argatroban (32 microG/kg/h, MD-group). During a 5-h observation period, the anti-Xa activity in circulating blood between the UFH- and DX-group and the anti-thrombin activity between the UFH- and MD-group were not significantly different. The graft patency in the DX-group (4/4) and MD-group (4/4) was significantly better than that in the UFH-group (3/10). Ultrastructural analysis of the luminal surface of the harvested graft by scanning electron microscopy revealed the reduced formation of fibrin networks entrapping erythrocytes in the DX- and MD-group in comparison with the patent UFH-group. In conclusion, a novel synthetic Xa inhibitor DX-9065a exerts a potent in vivo antithrombotic effect, which was comparable with argatroban, a synthetic thrombin inhibitor. PMID- 9225219 TI - New method for assaying free and total cholesterol in cultured cells by high pressure liquid chromatography. AB - We developed a simple, sensitive and accurate method for assaying cellular free and total cholesterol by monitoring 4-cholesten-3-one, a conversion product of the cholesterol oxidase-catalyzed oxidation of the free cholesterol that has a strong chromophoric alpha, beta-unsaturated ketone at 240 nm, using a high pressure liquid chromatographic system. This method measured picomole quantities of free and total cholesterol and precisely determined their concentrations in cells (10(4) range) in culture using 7 beta-hydroxycholesterol as an internal standard. PMID- 9225220 TI - Current state of and recent trends in serum lipid levels in the general Japanese population. Research Committee on Serum Lipid Level Survey 1990 in Japan. AB - To determine the recent serum lipid levels in the general Japanese population and trends in their changes over the past 30 years, a nationwide survey of serum lipid levels was conducted in 39 institutes from various districts around Japan. The total number of subjects were 34,815, consisting of 20,279 men and 14,536 women aged 4 through 99 years. All the serum samples were collected and analyzed within one week at the Special Research Laboratory (Tokyo, Japan). In males, the mean serum cholesterol level showed a gradually increase from 170 mg/dl in the 0- to 9-year-old age group to 198 mg/dl in the 50- to 59-year-old age group. There was a slight decrease after age 60 years. In females, the mean cholesterol level gradually rose with age from 173 mg/dl in the 0- to 9-year-old age group to 210 mg/dl in the 60- to 69-year-old age group, and fell to 207 mg/dl after 80 years of age. The mean HDL-cholesterol level in men gradually decreased with age from 60 mg/dl in the 0- to 9-year-old age group to 51 mg/dl in the 30- to 39-year-old age group, remained at this level up to 69 years of age, and then increased to 54 mg/dl for the above 80 years old group. The mean HDL-cholesterol level in women increased from 57 mg/dl in the 0- to 9-year-old age group to 62 mg/dl for the 20- to 29-year-old age group: then gradually decreased with age to 54 mg/dl in the 60 to 79-year-old age group. The mean LDL-cholesterol level in men gradually increased with age from 98 mg/dl in the 0- to 19-year-old age group to 122 mg/dl at 70-79 years of age. The mean LDL-cholesterol level in women was low at 101-103 mg/dl up to 29 years of age, then it increased with age to 135 mg/dl in the 60- to 69-year-old age group. The serum cholesterol levels in 1970 and 1980 were higher than that in 1960, but in 1990 values similar to those in 1960 were observed in both men and women. The present results will become the standard serum lipid level data for the Japanese people, and succeeding 10-year surveys will clarify the trends of lipid levels in this country. PMID- 9225222 TI - Lipoprotein (a) in the regulation of fibrinolysis. AB - Elevated plasma levels of lipoprotein(a) [LP(a)] are associated with increased an risk of developing atherosclerosis. This increased risk may be due to an Lp(a) mediated depression of fibrinolytic activity. Lp(a) regulates fibrinolysis by controlling the activity of plasminogen activators. Lp(a) is a low density lipoprotein with an apoprotein(a) subunit which has a high degree of homology with the fibrinolytic zymogen plasminogen. The apoprotein(a) subunit contains up to thirty seven copies of a domain homologous to the plasminogen kringle 4 domain, which enables Lp(a) to bind to fibrin. The subunit also has a zymogen domain, but it is not activated by plasminogen activators. Lp(a) inhibits plasminogen activation by competing with plasminogen for access to plasminogen activators bound to vascular surfaces. Lp(a) also competes with the irreversible inhibitor of plasminogen activators, plasminogen activator inhibitor-1. Therefore increases in Lp(a) concentration may decrease fibrinolytic activity by preventing activation of plasminogen, but Lp(a) may also prolong plasminogen activation by preventing the irreversible inhibition of the activators. At elevated levels of Lp(a) the decreased rate of plasmin generation may not be offset by the prolongation in plasminogen activation, and fibrinolysis will be inhibited. PMID- 9225221 TI - Cellular binding and degradation of lipoprotein (a). AB - Lp(a) is an important contributing factor to the development of atherosclerosis, and in structure is similar to LDL. Given the central role of the LDL receptor (LDL-R) in the metabolism of LDL, we felt that a study of the binding and degradation of Lp(a) facilitated by the LDL-R of human monocyte derived macrophages (HMDM) would be of value in understanding its pathological nature. In this study we compared equimolar amounts of Lp(a) and LDL and found that nearly equal amounts of Lp(a) and LDL bound to the LDL-R of HMDM at 4 degrees C, however the affinity of both lipoproteins was much lower than has been observed for the LDL-R of fibroblasts, being 0.80 muM for Lp(a) and 0.23 muM for LDL. The binding of Lp(a) to HMDM could be competed by 63% with a 50-fold excess of LDL. Degradation of Lp(a) at 37 degree C, unlike 4 degrees C binding, was mainly nonspecific (75% of total Lp(a) degradation) and when compared on an equimolar basis, nearly 6 times more LDL than Lp(a) was processed by the LDL-R pathway in 5 hr. Lower degradation of Lp(a) appears to be the result of lower binding at 37 degree C and a lower degradation rate when compared to LDL. It was not caused by increased intracellular accumulation or retroendocytosis. Degradation of both lipoproteins was only modestly affected by up and down regulation of the LDL-R. Because the binding of LDL at 4 degrees C and degradation at 37 degree C is mainly LDL-R specific, whereas only the 4 degree C binding of Lp(a) is so, suggests that the poor LDL-R dependent degradation of Lp(a) at 37 degree C is caused by a conformational change that is inducted in Lp(a) upon lowering the temperature to 4 degree C which allows better recognition of Lp(a) by the HMDM LDL-R. PMID- 9225223 TI - Polymorphism and Distribution of Apo(a). AB - Several apo(a) isoforms, controlled by a series of alleles Lp(a)F, Lp(a)B, Lp(a)S1, Lp(a)S2, Lp(a)S3, Lp(a)S4 and null, were found in 470 healthy Japanese by 4% SDS-PAGE and immunoblotting techniques. There was a strong inverse relationship between the apparent molecular weight of apo(a) isoforms and plasma concentrations of Lp(a). Lp(a) in d < 1.006 fraction increased 2-4h after oral fat load. Lp(a) exhibited a marked avidity for triglyceride-rich lipoprotein (TRL), and we suggest that the TRL-bound Lp(a) is the intact Lp(a) derived from serum. We demonstrated that the lipid-free apo(a) does not contain apo B-100 in serum, and has a molecular mass of ca 200 kDa. The free apo(a) level in normal subjects was 1.75 mg/dl (as Lp(a)) and was no different from the level in CAD patients. PMID- 9225224 TI - Lipoprotein (a) in ischemic heart disease and cerebrovascular disease. PMID- 9225225 TI - Changes in the concentration and distribution of lipoprotein (a) in plasma after fat intake. AB - To clarify the association between apo(a) and TG-rich lipoproteins, we studied changes in plasma Lp(a) concentration and apo(a) distribution in lipoprotein fractions after fat intake. The subjects were 15 hyperlipidemic patients and 3 healthy volunteers with fasting Lp(a) concentrations ranging from 2.5-52 mg/dl. They were given a fatty meal (50 g fat/m2 and 60 mg retinyl palmitate) after a 12 hour overnight fast and venous blood samples were taken at 0, 3, 4.5, 6, 7.5, 9, 12 and 24 hours. Fractions of sf > 400, sf 20-400 and d > 1.006 g/ml were isolated from plasma samples by ultracentrifugation. Plasma Lp(a) levels increased transiently at 4.5 hours, decreased between 4.5 and 12 hours, and recovered almost to initial levels by the next morning. Plasma Lp(a) peaked before the plasma RP peak appeared. Apo(a) associated with TG-rich lipoprotein fractions increased and apo(a) in the d > 1.006 fraction decreased after fat intake in most of the subjects, suggesting the transfer of apo(a) from the d > 1.006 fraction to the TG-rich lipoproteins. The increase in TG-rich lipoprotein apo(a) correlated with the RP area under the curve (r=0.79, p<0.05) and the decrease in d > 1.006 apo(a) (r = 0.80, p < 0.05). This distributional change of apo(a) after fat intake was confirmed by gel filtration and density gradient ultracentrifugation. Transfer of apo(a) from the main Lp(a) fraction of the plasma obtained from a subject with a high Lp(a)level to the TG-rich lipoprotein fraction of the plasma obtained 4.5 hours after fat intake from a subject with a low plasma Lp(a) level was also shown in vitro by density gradient ultracentrifugation. Our studies revealed a significant association between apo(a) and TG-rich lipoproteins in the postprandial. Further studies are necessary to clarify the pathophysiological role of Lp(a) in TG-rich lipoproteins. PMID- 9225226 TI - Contribution of Lp(a) to the occurrence of vascular diseases: correlation of several risk factors including diabetes mellitus. AB - We demonstrated that Lp(a) levels in patients with arteriosclerosis obliterans (21.4 +/- 2.5 mg/dl) and in patients with ischemic heart disease (17.2 +/- 0.8 mg/dl) are higher than those in controls (15.4 +/- 0.7 mg/dl) or healthy controls (11.3 +/- 1.1 mg/dl). Lp(a) levels in patients with these vascular diseases were especially higher when there were known atherosclerotic risk factors such as diabetes mellitus, hypercholesterolemia or hypertension, although Lp(a) levels in patients with these risk-positive group was not different from that of control. These results suggest that Lp(a) contributes to the development of atherosclerotic vascular diseases especially when known atherosclerotic risk factors are not present. We also investigated the case of thromboangiitis obliterans, which is believed to develop from nonatherosclerotic mechanisms, and found that Lp(a) levels were higher (26.5 +/- 9.6 mg/dl) in such patients. PMID- 9225227 TI - Significance of hypertriglyceridemia in the occurrence of ischemic heart disease. PMID- 9225228 TI - Role of apolipoprotein E in lipoprotein metabolism and in the process of atherosclerosis. AB - Apolipoprotein E (apoE) plays an important role in plasma lipoprotein metabolism through its high affinity binding to cell surface low density lipoprotein (LDL) receptor. To determine the role of apoE in plasma lipoprotein metabolism, transgenic mouse lines with integrated rat apoE gene under control of metallothionein promotor were established. The plasma level of rat apoE in homozygotes for the transgene was 17.4 mg/dl after zinc induction. In this group, plasma cholesterol and triglycerides levels were 43%, 68% reduced as compared with controls, respectively. Gel filtration chromatography showed that lipid reduction was mainly due to decreased both very low density lipoproteins (VLDL) and LDL. Furthermore, we studied the effects of apoE on the atherogenic process in Watanabe heritable hyperlipidemic (WHHL) rabbits. We administered 10 mg of purified apoE intravenously into five WHHL rabbits three times a week from their age of 2.5 months to 11 months for 8.5 months. After sustained administration of apoE, we found a significant reduction in the accumulation of cholesterol ester in aortae (1.55 +/- 0.07 mg/g tissue) as compared to control rabbits (4.32 +/- 0.61 mg/g tissue). Thus, apoE plays an important role not only in plasma lipoprotein metabolism but also in atherosclerotic process. PMID- 9225229 TI - Postprandial lipoprotein metabolism in diabetes mellitus and obesity. PMID- 9225230 TI - Hyperlipidemia and hemostatic system. AB - The plasma levels of blood coagulation and fibrinolytic factors and the serum levels of lipids were measured in 62 subjects (22 normolipidemia and 40 hyperlipidemia) to investigate whether hyperlipidemia may affect the hemostatic system. Prothrombin, factors VII, IX and X were elevated in hyperlipidemic patients. The positive correlations were found between factors VII, IX and X, and triglyceride. The significant correlations were also found between VII and IX, and total cholesterol. Plasma levels of thrombin-antithrombin III complex (TAT), which reflects activation of coagulation system, were slightly but significantly higher in type IIb hyperlipidemia, although they were within normal range. Plasma levels of active plasminogen activator inhibitor (PAI) in type IIb and IV were significantly higher than in normals. A significant correlation was found between active PAI and triglyceride (r = 0.76, p < 0.0001). After the administration of fat emulsion to 18 patients with various diseases, which induced artificial hypertriglyceridemia, PAI levels as well as triglyceride levels significantly increased. These results suggest that hypertriglyceridemia may increase the synthesis and/or release of PAI, inducing a hypofibrinolytic condition, which could lead to thrombosis. It has been established that lipoprotein (a) [Lp(a)], which has a molecular structure homology to plasminogen, impairs fibrinolysis by its competitive inhibition of adsorption of plasminogen to vascular endothelial surface and/or fibrin. We assayed plasma levels of Lp(a) and parameters of blood coagulation and fibrinolysis in 168 patients with type II diabetes mellitus and 48 normal controls. In the diabetics, the levels of Lp(a) as well as levels of tissue-type plasminogen activator (t-PA) antigen and PAI activity were significantly higher than normal controls. Furthermore, it was shown that Lp(a) had a weakly negative correlation with t-PA antigen in the diabetics. These results suggest that an elevated level of Lp(a) may decrease release of t-PA, although the underlying mechanism remains unsolved. PMID- 9225231 TI - Serum triglycerides and blood coagulation factors VII and X, and plasminogen activator inhibitor-1. AB - It has been suggested that impaired fibrinolytic-coagulation system, such as increased concentration of inhibitors to fibrinolysis or activators to coagulations, occasionally may play a role in the development of atherosclerotic vascular disease. In this study, we aimed to elucidate the relationship of serum lipids to fibrinolytic-coagulation system. The subjects studied were 190 outpatients at Kyorin University Hospital, 108 of whom were mostly hypertension, diabetes mellitus and hyperuricemia (Control), 59 of whom were coronary heart disease (CHD), 25 of whom were cerebrovascular disease (CVD). Blood samples were measured the levels of blood coagulation factors VII (F-VII) and X (F-X), and plasminogen activator inhibitor-1 (PAI-1) in these subjects, together with the concentrations of serum lipids. The serum levels of F-X was significantly higher in CHD subjects than in controls (111 +/- 19% vs 101 +/- 22%, p < 0.05). However, there was a no significant difference of F-VII among three groups. And we found that the levels of serum lipids, especially serum triglycerides showed a significant positive correlation between the concentrations of F-VII (r = 0.343, p < 0.01) and F-X (r = 0.513, p < 0.01), and PAI-1 (r = 0.528, p < 0.001) in CHD and CVD subjects. For this reason, 156 bank employee subjects were also admitted to this study (Bank employees). In bank employee subjects, the serum levels of triglycerides also showed a significant positive correlation with the levels of F VII (r = 0.321, p < 0.001), F-X (r = 0.254, p < 0.001) and PAI-1 (r = 0.420, p < 0.001). These data suggest that serum lipids, particularly triglycerides have a close relationship with thrombogenesis as evidenced by activated F-VII and F-X in the extrinsic coagulation system and also by elevated PAI-1 activities in fibrinolysis. Therefore, when we try to prevent the patients from CHD or treat them, we ought pay attentions not only to serum cholesterol or LDL-cholesterol for their atherogenic actions, but also to triglycerides because of their close correlation with extrinsic coagulation system and anti-fibrinolytic activities. The reduction of fibrinolytic capacity due to increased plasma levels of F-VII, X and PAI-1 may have importance in atherosclerotic vascular disease, particularly in patients with hypertriglyceridemia. PMID- 9225232 TI - Hypertriglyceridemia and fatty liver: clinical diagnosis of fatty liver and lipoprotein profiles in hypertriglyceridemic patients with fatty liver. AB - Fatty liver has prevailed by 14% in the healthy population of this country. The factors contributing genesis of fatty liver were gender (male), obesity, high alcohol consumption, glucose intolerance and hypertriglyceridemia. And hypertriglyceridemia seems to be the common underlying factor to all other causes. The mechanism for accumulation of triglycerides in the liver can be explained at least by increased HTGL activities and elevated apo A-II levels, a postulated co-factor of HTGL. An hypertriglyceridemic patients with fatty liver had the insulin resistance. PMID- 9225233 TI - Molecular disorders of cholesteryl ester transfer protein. AB - Plasma cholesteryl ester transfer protein (CETP) facilitates the transfer of cholesteryl ester (CE) from HDL to apolipoprotein B-containing lipoproteins and therefore is a key protein in the reverse cholesterol transport system. The importance of plasma CETP in lipoprotein metabolism has been highlighted by the discovery of CETP-deficient subjects with a marked hyper-HDL-cholesterolemia. The deficiency of CETP causes various abnormalities in the concentration, composition, and functions of high density and low density lipoproteins. The current review will focus on some of the recent knowledge on CETP with special reference to the biochemical and molecular biological aspects of CETP. Furthermore, detailed information will be presented regarding the lipoprotein abnormalities and molecular basis of CETP deficiency. PMID- 9225234 TI - Identification of two apolipoprotein variants, A-I Kaho (Asp 51-->Val) and A-I Lys 107 deletion. AB - We have identified two apolipoprotein (apo) A-I variants using isoelectric focusing gel electrophoresis: apo A-I Kaho which has a relative charge of +1 compared to normal apo A-I4, and apo A-I Nanakuma2 which has a relative charge of -1. Sequence analysis of PCR-amplified DNA from the proband of apo A-I Kaho revealed a single substitution of aspartic acid (GAC) for valine (GTC) at residue 51. Sequence analysis of PCR-amplified DNA from the proband of apo A-I Nanakuma2 revealed a three-base (AAG or AGA) deletion between bases 186 and 193 from the 5' end of exon 4 that leads to deletion of Lys 106 or 107. This mutation may be the same as that of apo A-I Marburg or A-I Munster-2 reported by Rall et al. (Rall SC Jr, Weisgraber KH, Mahley RW, Ogawa Y, Fielding CJ, Utermann G, Haas J, Steinmetz A, Menzel HJ, and Assmann G, J Biol Chem, 259: 10063-10070, 1984). Because of its unique sequence between 185 and 193 from the 5' end of exon 4 of apo A-I gene, we could not define whether AAG or AGA is deleted by DNA sequencing. PMID- 9225235 TI - A novel nonsense mutation in exon 1 and a transition in intron 3 of the lipoprotein lipase gene. AB - We examined the lipoprotein lipase (LPL) gene by single strand conformation polymorphism (SSCP) and by restriction fragment length polymorphism (RFLP) analysis in 106 patients with hypertriglyceridemia to screen for novel mutations and to study the contribution of LPL genetic defects in hypertriglyceridemia. We found a single incidence of a homozygous novel nonsense mutation (216G-->A; 14Tryptophan-->stop codon) in exon 1 and 6 cases heterozygous for a single transition (C-->T) at six bp upstream from splicing acceptor site of intron 3. These mutations were not found in 105 normolipidemic controls. The proband homozygous for the nonsense mutation in exon 1, a 74 year old woman, had mild hyperchylomicronemia and her post-heparin plasma showed no LPL protein. However, four heterozygous among family members did not demonstrate hypertriglyceridemia. The frequency of heterozygosity for the C-->T transition in intron 3 was significantly different from that in normolipidemic controls. Therefore, it was suggested that the mutation is involved in hypertriglyceridemia. All of the heterozygotes were men with 4 patients having impaired glucose tolerance or diabetes mellitus. These observations suggest that this polymorphism in intron 3 combined with other as yet undefined factors may be related to hypertriglyceridemia. PMID- 9225237 TI - The synergistic effect of elastase and hydrogen peroxide on vascular endothelial cell injury is due to the production of hydroxylradical in the endothelial cells. AB - Protease inhibitors such as aprotinin and urinastatin inhibited vascular endothelial cell injury induced by PMA-stimulated leukocytes, although their inhibitors did not suppress the production of active oxygen species released from leukocytes. On the other hand, in the presence of pancreas elastase (10 micrograms/ml), hydrogen peroxide (50 microM) caused severe injury of endothelial cells isolated from the bovine carotid artery (% specific 51Cr release, % SR = 42.9 +/- 3.3%), although the % SR elicited by elastase or hydrogen peroxide alone, respectively, was below 1%. Elastase and hydrogen peroxide acted synergistically on the injury of endothelial cells from the bovine carotid artery similarly to that in the endothelial cells isolated from the bovine coronary artery and human umbilical vein. Furthermore, elastase derived from both pancreas and leukocyte induced this synergistic action on endothelial cell injury. To clarify the mechanism of vascular endothelial cell injury induced by the combination of elastase and hydrogen peroxide, we examined the effects of various radical scavengers and protease inhibitors. Deferoxamine mesylate completely inhibited the endothelial cell injury, while protease inhibitors such as antitrypsin and macroglobulin had a protective effect. Pretreatment of endothelial cells with deferoxamine mesylate also protected against this cytotoxicity. These findings suggested that the synergistic effect of elastase and hydrogen peroxide on the endothelial cell injury is due to the production of hydroxylradical in the endothelium and that this synergistic action might be partially involved in the endothelial cell injury induced by activated leukocytes. PMID- 9225236 TI - Characterization of vitronectins in atherosclerotic lesions. AB - Vitronectin is one of the major extracellular matrix proteins that accumulates in atherosclerotic lesions. A monoclonal antibody (EMR1a/212D) specifically stained the extracellular regions in thickened intima which colocalized well with lipid deposition. The antigenic glycoprotein with a molecular weight of 66KDa was revealed to be rabbit vitronectin. When homogenates of WHHL rabbit atheroma were subjected to immunoblot analysis using EMR1a/212D, four molecules with molecular weight 66, 56, 50 and 47KDa were detected. To confirm whether these smaller immunopositive bands were derived from mature vitronectin, another monoclonal antibody (EMR1b/244H) recognizing the polypeptide region of vitronectin was prepared. All four molecules were detected by EMR1b/244H as well as by EMR1a/212D. Two smaller vitronectins (56KDa and 50KDa) were found in atherosclerotic lesions and increased markedly during the development of atherosclerosis. On the other hand, the vitronectin detected in normal rabbit aorta was mainly of the mature type, while 56KDa and 47KDa forms were not detected. The total amount of the four vitronectins in atherosclerotic lesions was 38.5 +/- 5.0 ng/mg wet weight tissue, a value approximately 9.5 fold higher than that found in normal aorta. In conclusions, we found massive accumulation of these vitronectins concomitant with atherosclerotic development in rabbit aorta. PMID- 9225238 TI - Coronary segmental responses to acetylcholine in patients with hypercholesterolemia. AB - We investigated coronary segmental response to intracoronary acetylcholine (ACh) in 19 patients with hypercholesterolemia and 18 patients with normal cholesterol levels. All patients had atypical and chest pain and angiographically normal coronary arteries. After baseline angiography, ACh (3 and 30 micrograms/min) was infused into the left coronary artery, followed by infusion of nitroglycerin. Percent changes in diameter of the proximal, middle, and distal segments of the left coronary arteries were measured by quantitative angiography. In the normocholesterolemic group, 3 micrograms/min of ACh produced significant coronary vasodilation in the distal segments (+8.2 +/- 2.6%, p < 0.005), while 30 micrograms/min did not cause any changes. In the hypercholesterolemic group, 30 micrograms/min of ACh caused significant coronary vasoconstriction in the middle and distal segments (-7.2 +/- 1.9% and -6.2 +/- 1.9%, p < 0.001 and p < 0.01, respectively), while 3 micrograms/min caused no changes. In each group, vasodilator responses to nitroglycerin in the middle and distal segments were significantly greater than those in the proximal segments (p < 0.001). Our results suggest that impaired endothelial function may be evaluated more effectively in the distal coronary segments in patients in the early stage of epicardial coronary atherosclerosis attributable to hypercholesterolemia. PMID- 9225239 TI - The relationships of testosterone, estradiol, dehydroepiandrosterone-sulfate and sex hormone-binding globulin to lipid and glucose metabolism in healthy men. AB - We investigated the relationships of plasma sex hormones (free testosterone; free T, estradiol; E2 dehydroepiandrosterone-sulfate; DHEA-S) and sex hormone-binding globulin (SHBG) levels to lipid and glucose metabolism cross-sectionally in 212 apparently healthy men aged from 18 to 59 years. A multiple linear regression analysis for lipid and glucose parameters with age, body mass index (BMI), percent body fat (%fat), waist to hip ratio (WHR), estimated maximal oxygen uptake (VO2max), alcohol and cigarette consumption, sex hormones, and SHBG, respectively, as independent variables, was performed. DHEA-S was indicated as one of the independent predictors of both high density lipoprotein cholesterol (HDL-C), with a positive relation, and of triglyceride and total cholesterol/HDL C ratio, with a negative relation, while SHBG was one of the predictors of both HDL-C, with a positive relation, and of fasting insulin, with a negative relation. The E2 level was found to be negatively related to both low density lipoprotein cholesterol and fasting blood glucose. These findings thus suggest that the higher levels of SHBG, DHEA-S and E2 within physiological ranges in healthy men may partially help to maintain a desirable profile of the plasma lipid and glucose metabolism. PMID- 9225240 TI - Altered bile acid metabolism related to atherosclerosis in alloxan diabetic rats. AB - Normal and alloxan diabetic rats were kept on a 0.25% cholesterol diet for 12 months and the changes in serum cholesterol levels, and fecal excretion of sterols and bile acids were examined to elucidate the influence of changes in bile acid metabolism on manifestations of hypercholesterolemia and development of atheromatous lesions. Diabetic rats fed the cholesterol diet showed increases in bile acid synthesis and in the cholic acid group/chenodeoxycholic acid group (CA/CDCA) ratio, and developed significant hypercholesterolemia and atheromatous lesions. In contrast, normal rats showed increased bile acids synthesis but a decreased CA/CDCA ratio after feeding with the cholesterol diet, and developed neither hypercholesterolemia nor atheromatous lesions. Fecal sterol excretion and the cholesterol/sitosterol ratio decreased in diabetic rats. Positive correlations were found between the cumulative serum cholesterol level and the atheromatous lesion area, and between the fecal CA/CDCA ratio and the serum cholesterol level, in the latter of which the correlation was higher in rats on the cholesterol diet than in those on the standard diet. These findings suggest that alteration of bile acid metabolism with increases in cholic acid synthesis and CA/CDCA ratio in diabetic rats enhances cholesterol absorption to produce significant hypercholesterolemia, which in turn leads to development of atheromatous lesions. PMID- 9225241 TI - Elucidation of the structure of constrained bicyclopeptides in solution by two dimensional cross-relaxation spectroscopy: amatoxin analogues. AB - The evaluation of peptide structures in solution is made feasible by the combined use of two-dimensional NMR in the laboratory (NOESY) and rotating frames (ROESY), and by the use of molecular dynamics calculations. The present paper describes how both the NMR method and molecular dynamics calculations were applied to very rigid synthetic bicycle peptides that are analogues of natural amatoxins. The NMR theory, which allows the estimate of interatomic distances between interacting nuclei, is briefly discussed. The experimental data were compared with those of known solid-state structures. Three amatoxin analogues have been examined. Of these, one is biologically active (S-deoxo gamma[R] OH-Ile3-amaninamide) and its structure in the solid state has recently been worked out. The second and third analogues (S-dexo-Ile3-Ala5-amaninamide and S-deoxo-D-Ile3-amaninamide, respectively) are inactive and their solid-state structures are unknown. The data presented confirm the authors previous hypothesis that lack of biological activity of S-deoxo-Ile3-Ala5-amaninamide is due to the masking of the tryptophan ring by the methyl group of L-Ala and not to massive conformational changes of the analogue. PMID- 9225242 TI - Preferred conformation of peptides rich in Ac8c, a medium-ring alicyclic C (alpha,alpha)-disubstituted glycine. AB - A complete series of terminally blocked, monodispersed homo-oligopeptides (to the pentamer level) from the sterically demanding, medium-ring alicyclic C (alpha,alpha)-disubstituted glycine 1-aminocyclooctane-1-carboxylic acid (Ac8c), and two Ala/Ac8c tripeptides, were synthesized by solution methods and fully characterized. The preferred conformation of all the oligopeptides was determined in deuterochloroform solution by IR absorption and 1H-NMR. The molecular structures of the amino acid derivative Z-Ac8c-OH, the dipeptide pBrBz-(Ac8c)2-OH and the tripeptide pBrBz-(Ac8c)3-OtBu were assessed in the crystal state by X-ray diffraction. Conformational energy computations were performed on the monopeptide Ac-Ac8c-NHMe. Taken together, the results obtained strongly support the view that the Ac8c residue is an effective beta-turn and helix former. A comparison is also made with the conformational preferences of alpha-aminoisobutyric acid, the prototype of C (alpha,alpha)-disubstituted glycines, and of the other members of the family of 1-aminocycloalkane-1-carboxylic acids (Acnc with n = 3, 5-7) investigated so far. The implications for the use of the Ac8c residue in peptide conformational design are considered. PMID- 9225243 TI - Solution synthesis of human midkine, a novel heparin-binding neurotrophic factor consisting of 121 amino acid residues with five disulphide bonds. AB - Human midkine (hMK), a novel heparin-binding neurotrophic factor consisting of 121 amino acid residues with five intramolecular disulphide bonds, was synthesized by solution procedure in order to demonstrate the usefulness of our newly developed solvent system, a mixture of dichloromethane or chloroform and trifluoroethanol. The final protected 121-residue peptide was assembled from two large fully protected intermediates, Boc-(1-59)-OH and H-(60-121)-OBzl, in CHL/TFE(3:1, v/v) using water-soluble carbodiimide in the presence of HOOBt as coupling reagents. After removal of the protecting groups by HF followed by treatment with Hg(OAc)2 in 50% acetic acid, the fully deprotected peptide was subjected to the oxidative folding reaction. The final product was confirmed to have the correct disulphide structure from its tryptic peptide mapping and to possess the same biological activities as those of the natural product. In order to clarify the active region of the hMK molecule, the N-terminal half domains [(1 59) and (60-121)] were also synthesized by the same procedure used for the hMK synthesis. The C-half domain was confirmed to show the full pattern of bioactivities except for the neuronal cell survival activity, while the N-half one showed much less activity in general. PMID- 9225244 TI - The total chemical synthesis of monocyte chemotactic protein-1 (MCP-1). AB - The affinity-based N (alpha)-amino protecting group tetrabenzo[a,c,g,i]fluorenyl 17 methoxycarbonyl (Tbfmoc) has been utilized as a hydrophobic probe to allow the simple, quick and highly effective isolation of a 76 residue cysteine-containing protein (MCP-1). The base-labile Tbfmoc group can be removed under very mild conditions, which preserve the thiol-containing protein in the reduced state. Oxidative folding was then used to furnish the biologically active beta-chemokine MCP-1. PMID- 9225245 TI - 3(10)-Helices, helix screw sense and screw sense reversal in the dehydro-peptide Boc-Val-delta Phe-Gly-delta Phe-Val-OMe. AB - The pentapeptide Boc-Val-delta Phe-Gly-delta Phe-Val-OME, containing two dehydro phenylalanine (delta Phe) residues, has been synthesized and its structure investigated. In the crystalline state, the molecule adopts a right-handed 3(10) helical conformation stabilized by two intramolecular hydrogen bonds between CO of Val1 and NH of delta Phe4, and between CO of delta Phe2 and NH of Val5, respectively. NMR measurements are consistent with the presence of 3(10)-helical structures also in acetonitrile and dimethylsulphoxide solution: the distances between backbone protons estimated from NOE connectivities are in overall agreement with those observed in the solid state; the chemical shifts of the amide protons show the smaller temperature coefficients for the NHs that in solid state are involved in intramolecular hydrogen bonds. The CD spectra in acetonitrile, chloroform, methanol and dimethylsulphoxide display exciton couplets of bands corresponding to the delta Phe electronic transition at 280 nm; the sign of the bands is consistent with the presence of helical structures having a prevalent left-handed screw sense. Addition of 1,1,1,3,3,3-hexafluoro propan-2-ol gives rise to the gradual appearance of a couplet of opposite sign, suggesting the helix reversal from left-handed sense to right-handed sense. The conformational behaviour is discussed on the basis of the specific sequence of the peptide. PMID- 9225247 TI - Synthesis of a new template with a built-in adjuvant and its use in constructing peptide vaccine candidates through polyoxime chemistry. AB - Synthetic lipopeptides are showing promise as vaccine candidates, but until now it has been very difficult to prepare them in homogeneous form. We describe the synthesis and characterization of a new water-soluble, four-branched template with a built-in lipophilic adjuvant (Pam3Cys). Through the use of oxime chemistry, we attached four copies of an unprotected influenza virus peptide and characterized the product (13 kDa) by reversed-phase HPLC and electrospray ionization mass spectrometry. Several other such constructions were made using the new template and different peptides. We seem to have a general method for making synthetic lipopeptides in homogeneous form. PMID- 9225246 TI - Crystal structure and molecular conformation of the cyclic hexapeptide cyclo-(Gly Aib-Gly)2. AB - We have synthesized and crystallized the cyclic peptide (Gly-Aib-Gly)2. Its structure has been determined by conventional X-ray diffraction methods. In the crystal it adopts a conformation with one beta-turn (type I) and its mirror image at the other side of the ring. All conformational angles are similar to those reported for these amino acids residues. In particular the Aib residue has a conformation intermediate between alpha- and 3(10)-helical conformations. The ring is an adequate model for the beta-turn conformation. A molecule of formic acid is found in the crystal which shows a very short hydrogen bond with one of the glycine carbonyl groups. PMID- 9225248 TI - The solution structure of the immunodominant and cell receptor binding regions of foot-and-mouth disease virus serotype A, variant A. AB - Abstract: The solution structure of a 20 amino acid long peptide corresponding to the region 141-160 of the envelope protein Vp1 from foot-and-mouth disease virus (FMDV) serotype A, variant A, has been determined by a combination of NMR experiments and computer calculations. The peptide contains both the immunodominant epitope as well as the sequence (RGD) used by the virus to bind the cell receptor in the initial stages of infection. These two sites have been shown to partially overlap. One hundred and thirty-five NMR distance constraints were used to obtain a set of 11 structures by distance geometry, minimization and molecular dynamics simulations. These structures were divided into two homogeneous families based upon backbone superimposition. The first and most populated family was characterized by a backbone RMS of 1.5 +/- 0.4 A, the second by a backbone RMS of 0.8 +/- 0.2 A. The two families had similar structural features and differed mainly in the backbone angles of G149. In the larger of the two families these angles favoured the formation of a loop comprising residues 147 to 152 and stabilized by a H-bond between NH of D147 and the CO of A152. In the second family, where this bond was absent, the peptide adopted in this region the shape of an irregular helix. The C-terminal half of the peptide (152-159) was similar in both families and largely helical. Similar structural features were also found within the VRGDS sequence (144-148) which was assigned to a beta-turn type IV. The features of the two families of structures were found to be different from those of the recently published X-ray structure of the antigenic loop of a chemically modified form of FMDV. Proposals accounting for these differences are provided which take into account the dual activity of the 141-160 sequence (i.e. antibody binding and cell invasion through receptor binding). PMID- 9225249 TI - The solution conformational features of two highly homologous antigenic peptides of foot-and-mouth disease virus serotype A, variant A and USA, correlate with their serological properties. AB - The solution structure of a peptide corresponding to the VP1 region 141-160 of foot-and-mouth disease virus (FMDV) serotype A variant USA has been studied by NMR and computer calculations and compared with the results from a study on a highly homologous peptide deriving from serotype A, variant A. The two peptides differ in their serological behavior and contain the immunodominant epitope of the virus which partly overlaps with its receptor binding region. Distance constraints, derived both from 2D and 3D homonuclear NMR and 2D-heteronuclear NMR experiments, were combined with DG calculations to yield 50 structures. After refinement through EM and restrained molecular dynamics simulations the selected structures shared several general features. In particular the 151-158 region was a helix in all cases while a large loop similar to that found in peptide A but comprising less residues and stabilized by an H-bond between the side chains of D147 and S150 was found in the majority of structures. A further loop, common to all structures, was identified around the RGD sequence (145-147). This was different from that found in the corresponding region of peptide A as were the conformations of the individual residues within the RGDX sequence. The different structural features shown by the two peptides were rationalized in terms of the S148 (peptide A) to F148 (peptide USA) mutation. The second mutation, that at position 153 (L in A, P in USA) did not appear to affect the structure of the peptide significantly although the different dimensions of the loop in the central region and the type of H-bond stabilizing it could be potentially ascribed to this second mutation. All criteria used pointed to different structural features for the two peptides consistent with their serological behaviour. PMID- 9225250 TI - Effects of end group and aggregation on helix conformation: crystal structure of Ac-(Aib-Val-Ala-Leu)2-Aib-OMe. AB - The role of end groups in determining stereochemistry and packing in hydrophobic helical peptides has been investigated using an alpha-aminosobutyric acid (Aib) containing model nonapeptide sequence. In contrast to the Boc-analogue, Ac-(Aib Val-Ala-Leu)2-Aib-OMe crystallizes with two independent molecules in a triclinic cell. The cell parameters are: space group P1, a = 10.100(2)A, b = 15.194(4)A, c = 19.948(5)A, alpha = 63.12(2) degrees, beta = 88.03(2) degrees, y = 88.16(2) degrees, Z = 2, R = 7.96% for 5140 data where magnitude of Fo > 3 rho(F). The two independent molecules alternate in infinite columns formed by head-to-tail hydrogen bonding. The helices in the two independent molecules are quite similar to each other but one molecule is rotated approximately 123 degrees about its helix axis with respect to the other. All the helical columns pack parallel to each other in the crystal. Replacement of the bulky Boc group does not lead to any major changes in conformation. Packing characteristics are also similar to those observed for similar helical peptides. PMID- 9225251 TI - The structures of the frenatin peptides from the skin secretion of the giant tree frog Litoria infrafrenata. AB - The granular dorsal glands of the giant tree frog Litoria infrafrenata contain five peptides including caerulein (a known neuropeptide), and four new peptides named franatins 1 (MH+ = 1140 Da), 2 (1423), 3 (2180), 4 (2493). The amino acid sequences of the frenatins are detailed: their structures do not correspond to those of peptides isolated from other amphibians or animals. Frenatin 3, Gly-Leu Met-Ser-Val-Leu -Gly-His-Ala-Val-Gly-Asn-Val-Leu-Gly-Gly-Leu-Phe-Lys-Ser-(OH), has wide spectrum antimicrobial properties. PMID- 9225252 TI - Solid-phase synthesis and cellular localization of a C- and/or N-terminal labelled peptide. AB - We report the solid-phase synthesis by the Fmoc strategy of a peptide containing a cysteamide group at its C-terminus. This peptide was subjected to further modifications including the linkage of fluorophores, namely lucifer yellow and coumarin respectively, at the C- and/or N-terminals. After incubation with living cultured cells these two probes were localized and it is concluded that the post synthesis modifications can strongly modify the localization of the peptide. PMID- 9225253 TI - Binding of Fe3+ ions to halobacterial purple membranes as studied by Mossbauer spectroscopy. AB - Purple membranes (PM) from Halobacterium were reconstituted with 57Fe ions and investigated by Mossbauer spectroscopy within the temperature range from 5 to 300 K at the Fe/bacteriorhodopsin (BR) ratio 0.6-300. When the Fe/Br ratio was below 2, Fe3+ bonded to PM mostly as hydroxymonomeric particle [FeOH]2+.5H2O, the apparent charge of the iron ion being two. When the Fe/BR ratio exceeded two, the dimeric form [FeOH](2+)4.8H2O along with a cluster form dominated. The temperature dependences of the mean square displacement show that the mobility of Fe ions changes from the solid-state type to the quasi-diffusional one at temperatures approximately 200 and approximately 230 K for the dimeric or monomeric and cluster iron forms, respectively. The nature of the cation binding sites and their location on the PM surface are discussed. A possible role of the divalent cation binding to PM in the mechanism of BR proton pumping is suggested. PMID- 9225254 TI - Polyclonal antibodies against human gamma-tubulin stain centrioles in mammalian cells from different tissues. AB - Rabbit polyclonal antibodies were raised against the C-terminal fragment (amino acid residues 318-451) of human gamma-tubulin. These antibodies were used to stain cultured cells of various tissues (epithelium, nervous tissue, fibroblasts) from different animals (human, monkey, pig, rat, kangaroo rat, mouse, hamster, chicken, triton). The antibodies specifically stained centrioles in the interphase and mitotic cells of mammals, but not birds (chicken) or amphibians (newt). In the interphase cells, centrioles were stained as a pair of dots (or as a double dot) in 96-97% of the cells. The distances between the maternal and filial centrioles varied in different cultures. Procentrioles were stained in certain cells, but with less intensity than mature centrioles. In mitotic cells, the antibodies revealed two spots corresponding to two mitotic poles. The spots in mitosis were significantly larger than the interphase dots, but the staining was more faint. In spontaneous tripolar mitoses, only two poles were stained. Thus, it was shown that, on the one hand, gamma-tubulin is associated with centrioles irrespective of whether or not they serve as the microtubule organizing centres and, on the other hand, gamma-tubulin might not be an essential component of the microtubule organizing centres. PMID- 9225255 TI - Effect of increasing concentrations of nonionic detergent Triton X-100 on solubilization and structure of rat liver and adipose plasma membranes. AB - The extent of solubilization and the structure of rat liver and adipose plasma membranes after treatment with nonionic detergent Triton X-100 were studied. The concentration of Triton X-100 varied from 0.005 to 0.050% (0.26-2.6 mg/mg membrane protein). The excimerization of the pyrene fluorescent probe and the relative vibronic band intensities in the pyrene monomer fluorescence spectrum were measured to evaluate the membrane lipid bilayer fluidity and polarity. The data on the aqueous pyrene fluorescence in the presence of Triton X-100 showed that formation of micellar aggregates occurred at detergent concentrations of over 0.015%. This value is a crucial factor in the detergent action on the membrane structure: both intensive extraction of plasma membrane proteins and fluidization of the lipid bilayer were observed only at Triton X-100 concentration over 0.015%. However, Triton X-100 did not exert any action on the polarity of the membrane hydrophobic regions. PMID- 9225256 TI - Effect of lysophosphatidylcholine on the structure and function of low density lipoproteins. AB - The treatment of LDL with bee-venom phospholipase A2 resulted in the formation of lipid-protein particles (phl-LDL) with an increased content of lysophosphatidylcholine (LPC). At the same time, the composition of other lipids and the protein structure remained unaffected. phl-LDL, as well as LPC, abolished the hormone-induced [Ca2+] increase in platelets and platelet aggregation induced by PAF, AMP and thrombin, whereas LDL produced no effect on the hormone stimulated increase in the intracellular [Ca2+]. The effect persisted in a Ca(2+) free medium, indicating that phl-LDL and LPC did not abolish the mobilization of intracellular stores with the above-mentioned inducers. Neither LPC no phl-LDL affected the [Ca2+]i level in platelets and suppressed the platelet aggregation evoked by tapsigargine, a specific inhibitor of endoplasmic reticulum Ca(2+) ATPase, or by phorbol myristate acetate. The inhibitory effect depended on the LPC concentration and the time of platelet incubation with phl-LDL or LPC. The half-maximum efficient LPC concentrations were identical for LPC and phl-LDL (2-4 microM). The inhibitory effect was dependent on the LPC structure: lysophosphatidylethanolamine and phosphatidylcholine displayed no inhibitory effect. The results suggest that when added to washed platelets, free LPC and phl LDL inhibit only the receptor-dependent increase of [Ca2+]i. PMID- 9225257 TI - Mixed lipid-protein films of bacterial photosynthetic reaction centres. II. Mixed multilayers on solid supports. AB - Mixed lipid-protein multilayers composed of the reaction centre (RC) proteins from the Chloroflexus aurantiacus and Rhodobacter sphaeroides (wild type) photosynthetic bacteria and synthetic lipids were investigated. The optimal conditions for forming thin films on solid plates (approximately 100% transfer) were 30 mN/m surface pressure and transfer of the interfacial monolayers from the buffer/air interface onto the plates by the Langmuir-Schaefer method. The films transferred onto quartz and optical transparent current-conducting plates retained their optical and photoelectric properties. The preferential orientation of the protein of the interfacial surface depended on the type of lipid used. In RC-acryloylphosphatidylethanolamine films, the H subunit of the RC from Rhodobacter sphaeroides was oriented toward the water phase, in contrast to RC diacetylenic acid films, in which the H-subunit was oriented toward the air phase. It is shown that RC can change their orientation in a monolayer, even to the opposite one, depending on the type of lipid matrix. PMID- 9225258 TI - Use of hydrated reversed micelles of surfactant in organic solvent for stabilization of individual oligomeric forms of uridine phosphorylase from Escherichia coli K-12. AB - The catalytic activity of uridine phosphorylase from Escherichia coli K-12 entrapped in hydrated reversed micelles of aerosol OT (AOT) in octane has been studied as a function of the degree of hydration of the micelles. It was shown that the catalytic activity of uridine phosphorylase reached maximum values at [H2O/[AOT] ratios equal to 8.4, 12.9, 16.1 and 18.6. Based on the sedimentation data the conclusion has been made that the maxima of the catalytic activity correspond to the monomeric, dimeric, trimeric and tetrameric forms of the enzyme. The measurements of the rate of the enzymatic reaction catalyzed by uridine phosphorylase entrapped in hydrated reversed micelles at various concentrations of AOT indicate that the monomeric enzyme form, in contrast to the trimeric and tetrameric forms, exhibits the membranotropic properties. PMID- 9225259 TI - Conformational analysis of amphotericin B molecule. AB - Conformational analysis of a cyclic molecule of a channel-forming antibiotic amphotericin B was conducted by the methods of molecular mechanics. A number of conformers differing in both hydroxyl group orientation and lactone ring conformation were obtained. The stable states of the conformers have close intrinsic energies. The conformational analysis supports the conclusion by Mazerski and Borowski (Biophys. Chem. 54:49-60 (1995)) on the flexibility of the amphotericin B lactone ring. PMID- 9225260 TI - Intracellular distribution and characteristics of Ca(2+)-transporting systems in cells of ciliate protozoan Tetrahymena pyriformis. AB - A significant digitonin-sensitive calcium pool has been shown to exist in Tetrahymena pyriformis cells. In the presence of exogenic energy sources, calcium released from the digitonin-sensitive pool can be accumulated by mitochondria or endoplasmic reticulum. The kinetic characteristics of the mitochondrial and reticulum systems of Ca2+ transport and their sensitivity to various inhibitors were studied. PMID- 9225262 TI - ATP-dependent potassium channel from rat liver mitochondria: inhibitory analysis, channel clusterization. AB - An inhibitor of potassium mitochondrial channels quinidine (0.1 mM) closed the ATP-sensitive potassium channels isolated from mitochondria and reconstituted into a bilayer lipid membrane. Inhibitors of cytoplasm membrane K-channels (ouabain, tetraethyl ammonium, and cesium ions) produced no effect on the ATP sensitive mitochondria K-channels; 0.5-2 microM glybenclamide did not affect the reconstituted channels, either. The multiplicity of jumps of conductivity at small concentration of the protein and the maintenance of ion selectivity at switching various levels of conductivity lead us to assume that the large channels are clusters of elementary channels. The average frequency of channel cluster switchovers between various levels of conductivity is much higher than at the elementary channel. PMID- 9225261 TI - Direct participation of phospholipids in transmembrane K(+)-transport. AB - The correlation between the characteristics of K(+)-transport across the mitochondrial and bilayer lipid membranes formed form mitochondrial lipids has been demonstrated. It has been shown that different modes of K(+)-efflux activation in mitochondria result in the appearance of K(+)-transporting phospholipid forms. The experiments described in this work suggest that current fluctuations similar to those observed for biological channels can be registered in unmodified bilayer lipid membranes containing no protein components. PMID- 9225263 TI - Quantitative evaluation of the kinetics of lipid probe redistribution between fusing cells. AB - Redistribution of a fluorescent lipid probe in a cell-fusion system was analyzed by solving numerically the time-dependent diffusion equations on two spheres connected by a cylindrical pore. The half-times of the process were calculated for different values of the diffusion coefficient Dp for a lipid probe in the pore. It was shown that the kinetics of the redistribution was relatively insensitive to the length and radius of the pore, whereas Dp significantly affected the redistribution rate constant. Based on the results of the numerical analysis, the minimal lag time between the start of the process and the appearance of experimentally measurable quantities of the probe in the acceptor cell was roughly estimated. PMID- 9225264 TI - A theoretical study of outermost skin layer electroporation. AB - Two models of electrohydration of stratum corneum (SC) have been developed. In the first model, the hydration of one interbilayer region is considered on the assumption that water molecules are adsorbed on the inhomogeneous surface of a bilayer and can interact, thereby lowering their energy on the surface. The dependence of the hydration degree on the voltage across the skin had been found. At certain parameter values the degree of hydration rapidly grows at certain voltage up to the magnitudes at which continuous water pathways appear. The second model has used the macroscopic approach which presumes water to be present in the interbilayer region as microdrops. The dependence of the hydration degree on voltage has been also found. At voltages of the order of tens of volts the obtained hydrations of interbilayer regions are sufficient to generate electroinduced hydrophilic pores in the SC lipid phase. Formation of tortuous continuous pathways for the transport of small ions is little probable because it requires voltages much higher than 100 V. We suggest that small ions pass the skin by the straight way through corneocytes and lipid bilayers at voltages of the order of tens of volts and higher. PMID- 9225265 TI - 5-HT receptor classification and nomenclature: towards a harmonization with the human genome. AB - Molecular biology has dramatically advanced our knowledge and understanding of receptors for 5-hydroxytryptamine (5-HT). The existence of multiple 5-HT receptors defined using traditional pharmacological and biochemical approaches has now been amply confirmed, but gene products encoding putative "new" 5-HT receptors have also been discovered. In some cases, the absence of suitably selective agonists and antagonists has hampered determination of a physiological role for these gene products. This makes their classification as formally recognised receptors premature. PMID- 9225266 TI - Inhibition of cAMP accumulation via recombinant human serotonin 5-HT1A receptors: considerations on receptor effector coupling across systems. AB - Inhibition of forskolin-stimulated cyclic AMP accumulation was measured in two stable HeLa cell lines HA6 and HA7 expressing different levels of recombinant human 5-HT1A receptors. These cells were studied previously to characterize another second messenger system activated by 5-HT1A receptors, i.e. calcium mobilization. The pharmacological characterization of the inhibition of cyclic AMP accumulation was made using agonists (5-HT, 8-OH-DPAT, buspirone, MDL 73005) and putative antagonists (SDZ 216-525, NAN-190, WAY-100135, pindolol, propranolol, WAY 100635). It is shown that 5-HT, 8-OH-DPAT, buspirone, MDL 73005 behaved as full (or nearly full) and potent agonists, whereas SDZ 216-525, NAN 190 and WAY-100135 displayed a limited (and similar) degree of intrinsic activity at human 5-HT1A receptors; on the other hand pindolol, propranolol and WAY 100635 behaved as "silent" antagonists. The effects were quantitatively and qualitatively very similar in both cells lines for all drugs tested, suggesting that the coupling between 5-HT1A receptors and inhibition of cyclic AMP accumulation in HeLa cells is very tight. There were, however, significant variations in both the level of agonism and the potency of a number of compounds when calcium mobilization and the inhibition of cyclic AMP accumulation were compared. Especially in HA7 cells which express lower receptor levels, a number of drugs failed to display agonism (e.g. buspirone or MDL 73005), whereas in HA6 cells they acted as partial agonists. Together, the data show that functional responses mediated by the same receptor can vary rather dramatically depending on receptor density and/or on the effector system involved. Interestingly, 5-HT1A receptor-mediated inhibition of adenylate cyclase activity measured in calf hippocampal membranes shows very similar degrees of potency and intrinsic activity for a number of compounds that have been tested on the inhibition of cyclic AMP accumulation in HeLa cells, suggesting that the very tight coupling observed in the recombinant system may apply to native 5-HT1A receptors. PMID- 9225268 TI - Role of the 5-HT1A receptor in development of the neonatal rat brain: preliminary behavioral studies. AB - Serotonin exerts an influence on the prenatal development of rat brain. However, later developmental times may be more applicable to the understanding of the role of serotonin in human developmental disorders. Therefore, the current study was undertaken to gain preliminary information on the postnatal effects of serotonin on rat brain development. As the 5-HT1A receptor has been shown to be involved in much of the developmental functions of serotonin, an agonist for this receptor, 8 hydroxy-DPAT (8-OH-DPAT), was used. Neonatal rat pups at three ages (postnatal days, PNDs) 3-10, 10-17 or 17-24) were injected daily with 1 mg/kg 8-OH-DPAT and evaluated for behavioral consequences. The youngest group showed accelerated incisor eruption and eye-opening, a possible consequence of 5-HT1A receptor interactions with epidermal growth factor (EGF). Behaviorally, the animals were more anxious. Animals treated from PND 10-17, showed no change in craniofacial development but showed greater behavioral maturity in measures of spontaneous alternation and activity in the open field. The oldest animals (PND 17-24) showed no behavioral alterations, suggesting that this time length is beyond the critical period for serotonin's influence in brain development. PMID- 9225267 TI - Association of short alleles of a VNTR of the serotonin transporter gene with anxiety symptoms in patients presenting after deliberate self harm. AB - The polymorphism of a variable number tandem repeat (VNTR) region of the serotonin transporter gene consists of three alleles containing, respectively, 9 (STin2.9), 10 (STin2.10) and 12 (STin2.12) copies of a repetitive element. The frequencies of the three alleles in 384 individuals presenting after deliberate self harm were the same as a group of 346 controls. However, ratings of anxiety (and state anger) were higher in those patients with genotypes including the shorter repetitive elements. The findings support the hypothesis that, in this group of patients with low rates of severe psychiatric disorder, allelic variation in the serotonin transporter gene may contribute to the expression of anxiety symptoms. PMID- 9225269 TI - Agonist and inverse agonist efficacy at human recombinant serotonin 5-HT1A receptors as a function of receptor:G-protein stoichiometry. AB - Membrane preparations were made from Chinese Hamster Ovary (CHO) cells expressing 1.6 and 4.2 pmol/mg of recombinant human 5-HT1A receptors, as determined by saturation binding with the selective antagonist, [3H]-S 15535 ([3H]-4 (benzodioxan-5-yl)]-(indan-2-yl)piperazine). There was no change in the number of G-proteins activated by the full agonist, serotonin (5-HT; approximately 1.1 pmol/mg in each preparation, measured by [35S]-GTP gamma S saturation binding), therefore increasing the receptor:G-protein ratio from approximately 1.4:1 (RGlow) to approximately 4:1 (RGhigh). Agonist efficacy was measured by stimulation of [35S]-GTP gamma S binding. The serotonergic agonist, eltoprazine, behaved as a partial agonist (Emax = 52.7%) at RGlow membranes but virtually as a full agonist (Emax = 93.2%) at RGhigh membranes, relative to 5-HT (= 100%). The latter exhibited a two-fold shift to the left in its concentration-response curve in RGhigh compared to RGlow membranes (P < 0.01). WAY 100,635 (N-?2-[4-(2 methoxyphenyl)-1-piperazinyl]ethyl?-N-(2-pyridinyl) -cyclo-hexane-carboxamide), did not alter [35S]-GTP gamma S binding from basal levels in either membrane preparation. In contrast, spiperone displayed inverse agonist activity, decreasing [35S]-GTP gamma S binding from basal levels by 17% in RGlow membranes but by 28% in RGhigh membranes. These data indicate that an increased receptor:G protein ratio (i) augments the potency of full agonists, (ii) increases the efficacy of partial agonists and (iii) increases the negative efficacy of inverse agonists at recombinant human 5-HT1A receptors. Furthermore, these data suggest that spiperone induces, or stabilises, a G-protein-coupled, but inactive conformation of the receptor. PMID- 9225270 TI - WAY100635-induced augmentation of the 5-HT-elevating action of citalopram: relative importance of the dose of the 5-HT1A (auto)receptor blocker versus that of the 5-HT reuptake inhibitor. AB - The elevation of extracellular 5-HT after systemic administration of 5-HT reuptake inhibiting drugs is strongly potentiated by agents capable of blocking 5 HT1A autoreceptors in the midbrain raphe. The present in vivo microdialysis study was aimed at assessing the relative importance of 5-HT reuptake inhibition versus 5-HT1A autoreceptor blockade in this interaction. Citalopram (0.5 or 5.0 mg/kg s.c.) dose-dependently increased dialysate 5-HT in the rat ventral hippocampus, maximally doubling the initial baseline values within 60 min after injection. The selective 5-HT1A receptor blocker, WAY100635 (0.01-0.3 mg/kg s.c.), further augmented, in a dose-dependent manner, the high-dose citalopram response (to approximately 4-5 x the pre-citalopram baseline). For comparison, the effect of low-dose (0.5 mg/kg s.c.) citalopram was mildly, but not significantly, potentiated by WAY100635 (0.3 mg/kg). WAY100635 given alone does not alter 5-HT under these conditions. The data confirm previous findings that 5-HT1A autoreceptor blockade enhances the citalopram-induced increase of extracellular 5 HT in the forebrain. To the extent the extracellular levels of 5-HT is a valid index, through 5-HT reuptake blockade appears to be the primary prerequisite for this interaction to occur. New drugs and/or treatment regimes based on the SSRI/5 HT1A autoreceptor blocker combination concept should, therefore, emphasize the former property. PMID- 9225271 TI - Effect of 5-HT1A receptor antagonists on citalopram-induced increase in extracellular serotonin in the frontal cortex, striatum and dorsal hippocampus. AB - The aim of the present study was to compare the effects of citalopram, either alone or combined with 5-HT1A receptor antagonists, on extracellular serotonin levels in brain regions innervated by the dorsal or median raphe nuclei. Using intracerebral microdialysis in awake rats with separate probes in the frontal cortex or dorsal hippocampus, we studied the ability of 8 mg/kg s.c. ( )penbutolol, a beta-adrenoceptor antagonist with antagonist action at 5-HT1A and 5-HT1B receptors, and 0.3 mg/kg s.c. WAY-100635, a selective 5-HT1A receptor blocker, to modify the effect of 1 and 10 mg/kg i.p. citalopram on extracellular serotonin. Both doses of citalopram had more effect on extracellular serotonin levels in the dorsal hippocampus than in the frontal cortex. The effect of 1 mg/kg citalopram was significantly potentiated by (-)penbutolol in the frontal cortex only, but a clear-cut potentiation of the effect of citalopram was seen in both regions at a dose of 10 mg/kg. The effect of 10 mg/kg citalopram was potentiated by WAY-100635 in the frontal cortex but not in the dorsal hippocampus. In a second set of experiments, the combined effect of WAY-100635 and citalopram was studied in the same rat implanted with vertical probes in the striatum and dorsal hippocampus. Citalopram (1 and 10 mg/kg i.p.) raised extracellular serotonin to a similar extent in both regions. However, 0.3 mg/kg s.c. WAY-100635 potentiated the effect of 10 mg/kg citalopram in the striatum but not in the dorsal hippocampus. The results suggest that only a combined blockade of 5-HT1A and 5-HT1B receptors potentiates the effect of citalopram on extracellular concentrations of serotonin in the dorsal hippocampus. The findings may be relevant in designing clinical trials aimed at enhancing the antidepressant action of selective serotonin re-uptake inhibitors by combining them with serotonin receptor antagonists. PMID- 9225272 TI - Autoreceptor antagonists enhance the effect of the reuptake inhibitor citalopram on extracellular 5-HT: this effect persists after repeated citalopram treatment. AB - The effect of repeated administration of the reuptake inhibitor citalopram (10 mg/kg s.c., b.i.d. for 14 days) or saline on extracellular 5-hydroxytryptamine (5 HT) and autoreceptor sensitivity was assessed using microdialysis in the frontal cortex (FCx) and dorsal hippocampus (DH) of unanesthetized rats. Acute citalopram (5 mg/kg s.c.) challenge produced significant increases in DH and FCx 5-HT. The nonselective 5-HT1A/1B receptor antagonist (-)+penbutolol (8 mg/kg s.c.), administered 2 hr after citalopram challenge, significantly enhanced 5-HT in FCx and DH of both the chronic citalopram and saline pretreatment groups. Administration of the selective 5-HT1A receptor antagonist WAY 100635 (0.3 mg/kg s.c.) after citalopram challenge significantly enhanced 5-HT in FCx but not DH of both pretreatment groups. This suggests that there may be differences between DH and FCx in regulation of 5-HT release. Nevertheless, these results provide evidence that 5-HT autoreceptors are still active in restraining 5-HT release. Nevertheless, these results provide evidence that 5-HT autoreceptors are still active in restraining 5-HT release even after repeated administration of an antidepressant drug. PMID- 9225273 TI - Effects of food deprivation on midbrain 5-HT1A autoreceptors in Lewis and SHR rats. AB - Food deprivation stimulates the activity of the hypothalamo-pituitary-adrenal axis and brain serotonin (5-hydroxytryptamine, 5-HT) synthesis. Because midbrain somato-dendritic 5-HT1A autoreceptors may obey homologous and heterologous (e.g. by glucocorticoids) down-regulation, we have analyzed whether 24 hr of fasting affects midbrain 5-HT1A receptor binding and sensitivity in Lewis and SHR rats (i.e. strains that differ in behavioral/neuroendocrine responses to stressors). Fasting affected neither [3H]8-hydroxy-2-(di-N-propylamino)tetralin ([3H]8-OH DPAT) binding at 5-HT1A autoreceptors nor 8-OH-DPAT-induced inhibition of midbrain 5-HT synthesis (an index of 5-HT1A autoreceptor sensitivity). Because fasting increased 5-HT precursor (tryptophan) levels to similar extents in the midbrains of saline- and 8-OH-DPAT-treated rats, we conclude that food deprivation does not affect 5-HT1A autoreceptors. In turn, our results suggest that the differential effects of 5-HT1A receptor agonists on food intake, in fed and fasted rats may be independent from 5-HT1A autoreceptors. PMID- 9225274 TI - In vivo effects of 5-hydroxytryptamine receptor activation on rat nucleus tractus solitarius neurones excited by vagal C-fibre afferents. AB - The effects of ionophoretically applied 5-hydroxytryptamine (5-HT) and 5-HT receptor agonists were studied on rat nucleus tractus solitarius (NTS) neurones receiving unmyelinated vagal afferent input. 5-HT excited 15 of 34 neurones (44%), inhibited 10 (29%) and had no effect on nine. 8-Hydroxy-2-(di-N propylamino)tetralin HBr (8-OH-DPAT) excited 23 of 53 neurones (43%), inhibited 24 (45%) and had no effect on six neurones and (+/-)-2,5-dimethoxy-4 iodoamphetamine HCl activated 18 of 37 neurones (49%), inhibited nine (24%) and had no effect on 10. These results demonstrate that activation of 5-HT1A and 5 HT2 receptors can excite or inhibit populations of NTS neurones. Phenylbiguanide, however, excited 20 of 23 neurones (87%), inhibited only one (4%) and had no effect on two indicating that 5-HT3 receptor activation has an excitatory action. NTS neurones receiving cardiac vagal afferent input were more likely to be excited by 5-HT (five of five, 100%) or 8-OH-DPAT (four of five. 80%) than the population as a whole. In conclusion, the data demonstrate that 5-HT1A, 5-HT2, and 5-HT3 receptor subtypes are functionally present on NTS neurones receiving excitatory vagal afferent input. Further, the subpopulation of NTS neurones receiving input from cardiac afferents are excited by 5-HT, possibly by an action on 5-HT1A or 5-HT3 receptors. PMID- 9225275 TI - How efficacious are 5-HT1B/D receptor ligands: an answer from GTP gamma S binding studies with stably transfected C6-glial cell lines. AB - The intrinsic activity of a series of 5-hydroxytryptamine (serotonin, 5-HT) receptor ligands was analysed at recombinant h5-HT1B and h5-HT1D receptor sites using a [35S]GTP gamma S binding assay and membrane preparations of stably transfected C6-glial cell lines. Compounds either stimulated or inhibited [35S]GTP gamma S binding to a membrane preparation containing either h5-HT1B or h5-HT1D receptors. The potencies observed for most of the compounds at the h5 HT1B receptor subtype correlated with their potencies measured by inhibition of stimulated cAMP formation on intact cells. Apparent agonist potencies in the [35S]GTP gamma S binding assay to C6-glial/h5-HT1D membranes were, with the exception of 2-[5-[3-(4-methylsulphonylamino)benzyl-1 2,4-oxadiazol-5-yl]-1H indol-3-yl] ethanamine (L694247), 5- to 13-times lower than in the cAMP assay on intact cells. This suggests that receptor coupling in the h5-HT1D membrane preparation is less efficient than that in the intact cell. It further appeared that 6-times more h5-HT1D than h5-HT1B binding sites were required to attain a similar, maximal (73%), 5-HT-stimulated [35S]GTP gamma S binding response: Hence, the h5-HT1B receptor in C6-glial cell membranes could be more efficiently coupled, even though some compounds more readily displayed intrinsic activity at h5-HT1D receptor sites [e.g. dihydroergotamine and (2'-methyl-4'-(5 methyl[1,2,4]oxadiazol-3-yl)biphenyl-4-carboxylic acid [4-methoxy-3-(4 methylpiperazin-1-yl)phenyl]amide (GR127935)]. Efficacy differences were apparent for most of the compounds (sumatriptan, zolmitriptan, rizatriptan, N-methyl-3 [pyrrolidin-2(R)-ylmethyl]-1H-indol-5-ylmethyl sulfonamide (CP122638), dihydroergotamine, naratriptan and GR127935) that stimulated [35S]GTP gamma S binding compared to the native agonist 5-HT. The observed maximal responses were different for the h5-HT1B and h5-HT1D receptor subtypes. Few compounds behaved as full agonists: L694247, zolmitriptan and sumatriptan did so at the h5-HT1B receptor and only L694247 at the h5-HT1D receptor. GR127935 (10 microM) exerted little effect on [35S]GTP gamma S binding via h5-HT1B receptors (10% stimulation), but potently (pA2: 9.11) antagonized h5-HT1B receptor-stimulated [35S]GTP gamma S binding. Ketanserin and methiothepin inhibited [35S]GTP gamma S binding (by 13-28%) in the absence of an agonist, but were potent and competitive antagonists in the presence of an agonist via h5-HT1B (methiothepin) and h5-HT1D (methiothepin and ketanserin) receptors. The results document the utility of using [35S]GTP gamma S binding studies to assess agonist efficacy, and to characterize 5-HT1B/D receptor ligands as apparently neutral antagonists and inverse agonists at the G-protein level. PMID- 9225276 TI - Molecular cloning and pharmacological characterization of guinea pig 5-HT1B and 5 HT1D receptors. AB - Human 5-HT1B and 5-HT1D receptors have been implicated as molecular targets for the treatment of acute migraine based upon the pharmacological actions and clinical efficacy of sumatriptan, an agonist for human 5-HT1B/1D receptors. The guinea pig has served as an animal model to assess 5-HT1B/1D receptor function, most recently in evaluating 5-HT1B/1D receptor agonists as potential anti migraine agents. Since two distinct, but closely-related receptors displaying "5 HT1D receptor pharmacology" have been cloned previously from most mammalian species, the genes encoding these receptors were isolated from a guinea pig liver genomic DNA library using oligonucleotide probes targeted to nonconserved regions of recombinant human 5-HT1B and 5-HT1D receptors. Sequence analysis indicates that guinea pig 5-HT1B and 5-HT1D receptors are comprised 390 and 378 amino acids, respectively. Comparison of the deduced amino acid sequences of guinea pig 5-HT1B and 5-HT1D receptor subtypes show that they display overall and transmembrane (TM) identities of 63% and 77%, respectively. Both clones contain a conserved threonine residue in TM7, a structural feature imparting "5-HT1D receptor pharmacology". Guinea pig 5-HT1B and 5-HT1D receptor genes were transiently expressed in Cos-7 cells and their binding properties were evaluated using [3H]5-HT. Both cloned receptor subtypes displayed "5-HT1D receptor pharmacology" with the following rank order of binding affinities: 5-CT > 5-HT > sumatriptan > 8-OH-DPAT > (-)-pindolol. Ketanserin displayed modest (five-fold) 5 HT1D receptor selectivity, while methiothepin exhibited a similar selectivity for the 5-HT1B subtype. In particular, ketanserin exhibits profound differences in 5 HT1D receptor affinity (and selectivity) across species. High correlations were observed between the binding affinities of serotonergic ligands for 5-HT1D binding sites measured in guinea pig cortical membranes and both cloned guinea pig 5-HT1B (r2 = 0.88) and 5-HT1D (r2 = 0.80) receptors, indicating that the development of subtype selective compounds (i.e. 5-HT1B versus 5-HT1D) using native tissues may be more difficult to achieve without the advantage of using recombinant receptor subtypes. Additionally, there is a good correspondence between binding profiles of recombinant guinea pig 5-HT1B and 5-HT1D receptor subtypes and to their respective cloned human homologs. However, species differences in binding affinities of a subset of compounds are evident. These data extend previous observations that subtype selective (i.e. 5-HT1D) compounds identified in one species may not discriminate between closely related receptors (i.e. 5-HT1B and 5-HT1D) in all animal model systems. PMID- 9225277 TI - Differential effects of 5-HT1B/1D receptor agonists on neurogenic dural plasma extravasation and vasodilation in anaesthetized rats. AB - These studies compared the effects of the 5-HT1B/1D receptor agonists sumatriptan, CP-122 288 ((R)-N-methyl-[3-(1-methyl-2-pyrrolidinylmethyl)-1H-indol 5-yl] methanesulphonamide succinate) and CP-93 129 (3-(1,2,5,6-tetrahydropyrid-4 yl)pyrrolo[3,2-b]pyrid-5-one dihydrochloride) on neurogenic dural extra-vasation and vasodilation in anaesthetized rats. Dural extravasation, evoked by high intensity (1.2 mA) stimulation of the trigeminal ganglion, was measured using the radioactive plasma marker 125I-labelled bovine serum albumin. Dural vasodilation produced by lower intensity (50-300 microA) stimulation of trigeminal fibres, was measured through a closed cranial window using intravital microscopy. All compounds inhibited dural extravasation (rank order of potency: CP-122 288 > > sumatriptan > CP-93 129) and dural vasodilation (rank order of potency: CP-93 129 > > sumatriptan = CP-122 288). Comparison of the potency of these compounds with their potencies in an in vitro functional model, agonist-induced [35S]GTP gamma S binding, suggests that blockade of dural extravasation was consistent with an action at rat 5-HT1D receptors, but activity at another, unknown, "extravasation receptor" could also be involved. In contrast, inhibition of dural vasodilation was consistent with an action at rat 5-HT1B receptors. We suggest that in our preparations, production of dural vasodilation involves activation of trigeminal A delta-fibres whereas production of dural extravasation involves activation of trigeminal C-fibres. The differential effects of compounds on dural extravasation and vasodilation may therefore be due to the different receptor subtypes involved and to the selective localization of these subtypes on different populations of trigeminal sensory fibre. PMID- 9225280 TI - Importance of h5-HT1B receptor selectivity for 5-HT terminal autoreceptor activity: an in vivo microdialysis study in the freely-moving guinea-pig. AB - The importance of h5-HT1B receptor selectivity for 5-HT terminal autoreceptor activity was investigated with the selective h5-HT1B receptor ligands SB 219085, SB 220272, SB 224289 and SB 216641. The studies employed measurement of compound affinity and efficacy in vitro and the measurement of extracellular 5-HT in the frontal cortex of the freely-moving guinea-pig using in vivo microdialysis. All compounds had high affinity and selectivity for the h5-HT1B receptor, with SB 224289 the most selective for h5-HT1B over h5-HT1D receptors. Compounds exhibited a range of efficacies at both receptors: SB 224289 and SB 219085 were inverse agonists, SB 220272 was an antagonist and SB 216641 was a partial agonist. SB 220272, SB 216641 and SB 224289 had no effect on extracellular 5-HT following systemic administration, however, SB 219085 produced a significant increase. The SB 219085-induced increase in extracellular 5-HT was attributed to the compounds non-specific releasing properties as it was also demonstrated to increase basal release of [3H]5-HT from pre-loaded guinea-pig cortical slices. The lack of effect of the above h5-HT1B receptor selective compounds and the decrease in extracellular 5-HT elicited by the non-selective compounds GR 127935, GR125743 and methiothepin suggest that antagonism of 5-HT1D receptors may mediate this decrease in 5-HT levels. It is plausible that blockade of 5-HT1D receptors increases 5-HT levels in the raphe, this activates 5-HTtA receptors which results in an overall decrease in terminal 5-HT release. Definitive proof now awaits elucidation of the action of a selective 5-HT1D receptor antagonist. PMID- 9225278 TI - Differential distribution of [3H]sumatriptan binding sites (5-HT1B, 5-HT1D and 5 HT1F receptors) in human brain: focus on brainstem and spinal cord. AB - We report on the autoradiographic distribution of 5-HT1B, 5-HT1D and 5-HT1F receptor subtypes in human brain, focusing on the brainstem and cervical spinal cord. We have used [3H]sumatriptan as a radioligand in the presence of suitable concentrations of 5-CT (5-carboxamidotryptamine) to define 5-HT1F receptors, and ketanserin, to discriminate between 5-HT1B and 5-HT1D receptors. In the brainstem the highest concentrations of [3H]sumatriptan binding sites were seen in substantia nigra. The spinal trigeminal nucleus, substantia gelatinosa of the spinal cord, nucleus of the tractus solitarius and periaqueductal grey, also showed significant levels of [3H]sumatriptan binding sites. In the brainstem and spinal cord the total population of 5-CT-insensitive receptors, corresponding to 5-HT1F receptors, ranged from 9.8% in the periaqueductal grey to 53.4% in the substantia gelatinosa. This population represented 67.0% of binding in layer V of the frontal cortex. The decrease in [3H]sumatriptan binding in the presence of 200 nM ketanserin, indicative of the presence of 5-HT1D receptors, was very limited throughout the human brain, only reaching 20% of total specific binding over the periaqueductal grey. The proportion of [3H]sumatriptan binding sites displaced by 5-CT and insensitive to ketanserin, corresponding to 5-HT1B receptors, was, in general, the most abundant, ranging from 43.8% in substantia gelatinosa to 69.9% in the periaqueductal grey. Significant levels of 5-HT1B and 5-HT1D receptors found in migraine control pain areas suggest their involvement in antinociceptive mechanisms. PMID- 9225279 TI - Identification of 5-HT receptor sub-types in a homogeneous population of presumptive serotoninergic neurones. AB - The expression of the known rat 5-HT receptor sub-types has been analyzed in a presumptive serotoninergic cell line derived from the rat raphe nuclei, using reverse transcription followed by polymerase chain reaction. By manipulating the activity of the oncogene (ts-SV40T) product used to immortalize the serotoninergic precursors, it has been possible to compare the expression of the 5-HT receptors in either replicative or differentiating cells. 5-HT1B, 5-HT1D, 5 HT3, 5-HT6 and 5-HT7 receptor gene expression were all observed in the replicating cells. However, under differentiation conditions, expression of all except the 5-HT1B receptor was lost. Only one novel amplification product appeared during early differentiation, in the 5-HT2B lane; its smaller than expected size was suggestive of a previously undescribed alternate splicing of the mRNA in brain. The curtailment of 5-HT receptor expression in differentiating neurones in vitro may reflect the normal ongoing restriction in the phenotypic potential during embryogenesis in vivo. The serotonin cells, therefore, constitute a pristine cell line in which to study the receptor pharmacology of one or more 5-HT receptor sub-types in isolation. PMID- 9225281 TI - Do 5-HT1B/1D autoreceptors modulate dorsal raphe cell firing? In vivo electrophysiological studies in guinea pigs with GR127935. AB - GR127935 is a selective antagonist of release-modulating 5-HT1B/1D autoreceptors on serotonergic terminals and, as such, would be expected to produce increases in extracellular 5-HT. The changes in 5-HT observed are mixed, however, possibly due to the presence of somatodendritic 5-HT1a/1D autoreceptors. Theoretically, blockade of these autoreceptors would elevate intra-raphe 5-HT which, in turn, would activate somatodendritic 5-HT1A autoreceptors and slow firing rate. As recorded in anesthetized guinea pigs, dorsal raphe cell firing was unaffected by doses of GR127935 ranging from 20 to 20000 micrograms/kg i.v. Lower doses of GR127935 (0.002-2 micrograms/kg i.v.) yielded highly variable responses, although these were not significantly different from baseline. 8-OH-DPAT in these and similar neurons produced the robust dose-dependent inhibitory response expected of a 5-HT1A agonist; increases in extracellular 5-HT resulting from re-uptake blockade by fluoxetine also suppressed unit activity. Doses of CP-135,807, a centrally-acting 5-HT1B/1D agonist, to increase tone on the somatodendritic 5 HT1B/1D autoreceptor produced only a trend toward decreases in dorsal raphe neuronal firing. The overall weak effect of GR127935 on raphe unit activity suggests that the mechanism described previously must take into account factors such as the degree of intra-raphe 5-HT release, the endogenous tone on the autoreceptors, receptor selectivity and intrinsic activity of GR127935 and/or heterogeneity within the subtype. PMID- 9225282 TI - Cloning and characterization of the guinea pig 5-HT1F receptor subtype: a comparison of the pharmacological profile to the human species homolog. AB - The anti-migraine compound, sumatriptan, has been shown to have substantial affinity for the cloned human 5-HT1F receptor suggesting that, in addition to 5 HT1B/5-HT1D receptor subtypes, the 5-HT1F receptor may be a therapeutic target for the treatment of migraine. Several investigators have used the guinea pig plasma extravasation model to evaluate potential anti-migraine drugs. Since species differences in the pharmacology of serotonin receptors are well known, we compared the pharmacological profiles of the cloned human and guinea pig 5-HT1F receptors in order to validate the usefulness of the in vivo model in predicting anti-migraine activity of compounds targeted for humans. We have cloned the guinea pig 5-HT1F by homology to the human 5-HT1F receptor and evaluated its pharmacological profile using radioligand binding assays. The cloned guinea pig 5 HT1F gene exhibited 94% amino acid identity to the corresponding human homolog. High affinity (Kd approximately 10 nM) [3H]5-HT binding was detected to membranes obtained from Cos-7 cells transiently expressing the guinea pig 5-HT1F receptor. The cloned guinea pig receptor displayed typical 5-HT1F receptor pharmacology with the following rank order of binding affinities: 5-HT > sumatriptan > 1-NP = DHE > alpha-methyl 5-HT > metergoline > methiothepin > 5-CT. The pharmacological profiles of the cloned guinea pig and human 5-HT1F receptors were very similar as reflected by the high correlation (r2 = 0.72, slope = 0.76) observed between the binding affinities of compounds for these two species homologs. In situ hybridization studies in guinea pig tissue revealed 5-HT1F receptor mRNA expression in the neurons of the trigeminal ganglion, suggesting that the 5-HT1F receptor may play a role in the presynaptic inhibition of neuropeptide release at the level of the intracranial vasculature, thereby blocking the development of neurogenic inflammation. Dorsal root ganglion cells also moderately expressed the 5-HT1F transcripts. The localization of the 5-HT1F receptor to areas involved in the mediation and transfer of nociceptive information implies a role for this receptor in pain processing. These findings indicate that a selective 5-HT1F agonist may be a novel approach to treat migraine. PMID- 9225283 TI - In vivo electrophysiological characterization of 5-HT receptors in the guinea pig head of caudate nucleus and orbitofrontal cortex. AB - The aim of the present study was to characterize in vivo the 5-HT receptor subtypes which mediate the effect of microiontophoretic applied 5-HT in the guinea pig head of caudate nucleus and orbitofrontal cortex. 5-HT and the preferential 5-HT2A receptor agonist DOI and the preferential 5-HT2C receptor agonist mCPP, suppressed the quisqualate (QUIS)-induced activation of neurons in both structures. The inhibitory effect of DOI and mCPP was not prevented by acute intravenous administration of the 5-HT1/2 receptor antagonist metergoline (2 mg/kg) and the 5-HT2A/2C receptor antagonist ritanserin (2 mg/kg) in the two regions nor by the selective 5-HT2A receptor antagonist MDL100907 (1 mg/kg) in the head of caudate nucleus. However, the inhibitory effect of DOI, but not that of mCPP, was antagonized by a 4-day treatment with metergoline and ritanserin (2 mg/kg/day; using minipumps implanted subcutaneously) in head of caudate nucleus, but not in orbitofrontal cortex. Microiontophoretic ejection of the 5-HT1A/7 receptor agonist 8-OH-DPAT and of the 5-HT1A receptor antagonist WAY100635 both suppressed the spontaneous and QUIS-activated firing activity of orbitofrontal cortex neurons. At current which did not affect the basal discharge activity of the neuron recorded, microiontophoretic application of WAY100635 and BMY7378 failed to prevent the inhibitory effect of 8-OH-DPAT. The inhibitory effect of gepirone, which is a 5-HT1A receptor agonist but devoid of affinity for 5-HT7 receptors, was also not antagonized by WAY100635. Altogether, these results suggest the presence of atypical 5-HT1A receptors in the orbitofrontal cortex. The present results also indicate that the suppressant effect of DOI may be mediated by 5-HT2A receptors in head of caudate nucleus and atypical 5-HT2 receptors in orbitofrontal cortex. PMID- 9225284 TI - Serotonin induces excitatory postsynaptic potentials in apical dendrites of neocortical pyramidal cells. AB - By intracellular and whole cell recording in rat brain slices, it was found that bath-applied serotonin (5-HT) produces an increase in the frequency and amplitude of spontaneous excitatory postsynaptic potentials/currents (EPSPs/EPSCs) in layer V pyramidal cells of neocortex and transitional cortex (e.g. medial prefrontal, cigulate and frontoparietal). The EPSCs were suppressed by LY293558, an antagonist selective for the AMPA subtype of excitatory amino acid receptor, and by two selective 5-HT2A receptor antagonists, MDL 100907 and SR 46349B. In addition, the EPSCs were suppressed by the fast sodium channel blocker tetrodotoxin (TTX) and were dependent upon external calcium. However, despite being TTX-sensitive and calcium dependent, there was no evidence that the EPSPs resulted from an increase in impulse flow in excitatory neuronal afferents to layer V pyramidal cells. The EPSCs could be induced rapidly by the microiontophoresis of 5-HT directly to "hot spots" within the apical (but not basilar) dendritic field of recorded neurons, indicating that excitatory amino acids may be released by a TTX-sensitive focal action of 5-HT on a subset of glutamatergic terminals in this region. Consistent with such a presynaptic action, the inhibitory metabotropic glutamate receptor agonist (1S,3S) aminocyclopentane-1,3-dicarboxylate markedly reduced the induction of EPSPs by 5 HT. Postsynaptically, 5-HT enhanced a subthreshold TTX-sensitive sodium current, potentially contributing to an amplification of EPSC amplitudes. These data suggest 5-HT. via 5-HT2A receptors, enhances spontaneous EPSPs/EPSCs in neocortical layer V pyramidal cells through a TTX-sensitive focal action in the apical dendritic field which may involve both pre- and postsynaptic mechanisms. PMID- 9225285 TI - Activation of 5-HT2B receptors in the medial amygdala causes anxiolysis in the social interaction test in the rat. AB - In a recent study, we reported the presence of neurones expressing 5-HT2B receptor protein in the medial amygdaloid nucleus of the adult rat brain. In the present study, bilateral micro-injection of the 5-HT2B receptor agonist 1-[5-(2 thienylmethoxy)-1H-3-indolyl]propan-2-amine hydrochloride (BW 723C86, 0.09 and 0.93 nmol, 5 min pretest) into the medial amygdaloid nuclei increased the total interaction time of a pair of male rats in the social interaction test, to a comparable extent to chlordiazepoxide (5 mg/kg p.o., 1 hr pretest) without altering locomotor activity; indicative of anxiolytic activity. The increase in social interaction was prevented by pretreatment with the 5-HT2C/2B receptor antagonist N-(1-methyl-5-indoyl)-N'-(3-pyridyl) urea hydrochloride (SB 200646A, at 2 but not 1 mg/kg p.o., 1 hr pretest), which did not alter behaviour when given alone. Intra-amygdala BW 723C86 (0.09, 0.31 and 0.93 nmol, 5 min pretest) did not significantly alter the number of punished responses made when the same rats were examined seven days later in a Vogel punished drinking test, although chlordiazepoxide (5 mg/kg p.o., 1 hr pretest) produced the expected anxiolytic profile. The results are consistent with the proposal that activation of 5-HT2B receptors in the medial amygdala induces anxiolysis in the social interaction model but has little effect on behaviour in a punished conflict model of anxiety. These data suggest that serotonergic neurotransmission in this nucleus may selectively affect specific kinds of anxiety generated by different animal models. PMID- 9225286 TI - SB 242084, a selective and brain penetrant 5-HT2C receptor antagonist. AB - SB 242084 has a high affinity (pKi 9.0) for the cloned human 5-HT2C receptor and 100- and 158-fold selectivity over the closely related cloned human 5-HT2B and 5 HT2A subtypes respectively. SB 242084 had over 100-fold selectivity over a range of other 5-HT, dopamine and adrenergic receptors. In studies of 5-HT-stimulated phosphatidylinositol hydrolysis using SH-SY5Y cells stably expressing the cloned human 5-HT2C receptor, SB 242084 acted as an antagonist with a pKb of 9.3, which closely resembled its corresponding receptor binding affinity. SB 242084 potently inhibited m-chlorophenylpiperazine (mCPP, 7 mgkg i.p. 20 min pre-test)-induced hypolocomotion in rats, a model of in vivo central 5-HT2C receptor function, with an ID50 of 0.11 mg/kg i.p., and 2.0 mg/kg p.o. SB 242084 (0.1-1 mg/kg i.p.) exhibited an anxiolytic-like profile in the rat social interaction test, increasing time spent in social interaction, but having no effect on locomotion. SB 242084 (0.1-1 mg/kg i.p.) also markedly increased punished responding in a rat Geller-Seifter conflict test of anxiety, but had no consistent effect on unpunished responding. A large acute dose of SB 242084 (30 mg/kg p.o.) had no effect on seizure susceptibility in the rat maximal electroshock seizure threshold test. Also, while SB 242084 (2 and 6 mg/kg p.o. 1 hr pre-test) antagonized the hypophagic response to mCPP, neither acute nor subchronic administration of the drug, for 5 days at 2 or 6 mg/kg p.o. twice daily, affected food intake or weight gain. The results suggest that SB 242084 is the first reported selective potent and brain penetrant 5-HT2C receptor antagonist and has anxiolytic-like activity, but does not possess either proconvulsant or hyperphagic properties which are characteristic of mutant mice lacking the 5-HT2C receptor. PMID- 9225287 TI - RS-102221: a novel high affinity and selective, 5-HT2C receptor antagonist. AB - The 5-HT2C receptor is one of three closely related receptor subtypes in the 5 HT2 receptor family. 5-HT2A and 5-HT2B selective antagonists have been described. However, no 5-HT2C selective antagonists have yet been disclosed. As part of an effort to further explore the function of 5-HT2C receptors, we have developed a selective 5-HT2C receptor antagonist, RS-102221 (a benzenesulfonamide of 8-[5-(5 amino-2,4-dimethoxyphenyl) 5-oxopentyl]-1,3,8-triazaspiro[4.5]decane-2,4-dione). This compound exhibited nanomolar affinity for human (pKi = 8.4) and rat (pKi = 8.5) 5-HT2C receptors. The compound also demonstrated nearly 100-fold selectivity for the 5-HT2C receptor as compared to the 5-HT2A and 5-HT2B receptors. RS-102221 acted as an antagonist in a cell-based microphysiometry functional assay (pA2 = 8.1) and had no detectable intrinsic efficacy. Consistent with its action as a 5 HT2C receptor antagonist, daily dosing with RS-102221 (2 mg/kg intraperitoneal) increased food-intake and weight-gain in rats. Surprisingly, RS-102221 failed to reverse the hypolocomotion induced by the 5-HT2 receptor agonist 1-(3 chlorophenyl)piperazine (m-CPP). It is concluded that RS-102221 is the first selective, high affinity 5-HT2C receptor antagonist to be described. PMID- 9225288 TI - Effects of ketanserin on the discrimination of electrical stimulation of the dorsal raphe nucleus in rats. AB - The electrical stimulation of the dorsal raphe nucleus was used as a training cue in a discrimination paradigm. Sprague-Dawley rats were trained to discriminate between electrical stimulation (ES; 200 microA) of the dorsal raphe nucleus (DRN) and non-stimulation. This was accomplished by associating ES with intraperitoneal (i.p.) injection of lithium chloride (LiCl), following the session with electrical stimulation. This made the drinking of saccharin during ES aversive by conditioned taste aversion. Following training, rats decreased saccharin consumption in ES sessions. This discrimination was learned within three pairings of the ES with LiCl. Lowering the ES current to 50-100 microA resulted in levels of saccharin consumption similar to non-stimulation levels, whereas 150 microA showed a response intermediate between the stimulation response at 200 microA and non-stimulation. The discrimination of ES of the DRN (200 microA) was not affected by prior administration of the 5-HT2 antagonist ketanserin (1 or 2 mg/kg, i.p.), suggesting that activation of 5-HT2 receptors is not the primary discriminative cue generated by ES. However, the 5-HT2A/2C agonist DOI (0.25-0.5 mg/kg, i.p.), substituted for ES of the DRN, i.e. animals reduced saccharin consumption following DOI administration. This substitution of DOI for ES was antagonized by the administration of ketanserin (1 mg/kg, i.p.). These results suggest that ES of the DRN has properties that are similar to the activation of 5 HT2A/2C receptors by DOI. However, other stimuli such as activation of other 5-HT receptors are also involved, given that the discriminative cues of ES are not blocked by the 5-HT2A/2C antagonist ketanserin. PMID- 9225289 TI - Analysis of the ligand binding site of the 5-HT3 receptor using site directed mutagenesis: importance of glutamate 106. AB - The 5-HT3 receptor is a ligand-gated ion channel with significant structural similarity to the nicotinic acetylcholine receptor. Several regions that form the ligand binding site in the nicotinic acetylcholine receptor are partially conserved in the 5-HT3 receptor, presumably reflecting the conserved signal transduction mechanism. Specific amino acid differences in these regions may account for their distinct ligand recognition properties. Using site-directed mutagenesis, we have replaced one of these residues, glutamate 106 (E106), with aspartate (D), asparagine (N), alanine (A) or glutamine (Q) and characterized the ligand-binding and electrophysiological properties of the mutant receptors after transient expression in HEK-293 cells. The affinity for the selective 5-HT3 receptor antagonist [3H]GR65630 was decreased 14-fold in the mutant E106D (Kd = 3.69 +/- 0.32 nM) when compared to wildtype (WT, E106) 5-HT3 receptor (0.27 +/- 0.03 nM), while the affinity for E106N was unchanged (0.42 +/- 0.07 nM, means +/- SEM, n = 3-10). Decreased affinities for both E106D and E106N were observed for the antagonists granisetron, ondansetron and renzapride and for the agonists 5-HT (130- and 30-fold) and 2-methyl-5-HT (250- and 20-fold), respectively. Both mutants still formed 5-HT-activatable ion channels, but the high Hill coefficient of the concentration effect curves in wildtype (2.0) was decreased to unity in both cases. The EC50 of 5-HT was increased seven-fold in E106N (8.7 microM) when compared to wildtype (1.2 microM), but unchanged in E106D, and the potency of the antagonist ondansetron for both mutants was decreased. E106A and E106Q expressed poorly preventing a detailed characterization. These data suggest that E106 contributes to the ligand-binding site of the 5-HT3 receptor and may form an ionic or hydrogen bond interaction with the primary ammonium group of 5-HT. PMID- 9225290 TI - Allosteric potentiation of the 5-HT3 receptor-mediated ion current in N1E-115 neuroblastoma cells by 5-hydroxyindole and analogues. AB - Potentiation of the 5-HT3 receptor-mediated ion current in mouse N1E-115 neuroblastoma cells by 5-hydroxyindole (5-OHi) and three analogues (5 aminoindole, catechol and indole) was examined using whole-cell voltage clamp and single channel patch clamp techniques. The substances tested enhanced the amplitude of the maximum 5-HT-evoked ion current by 12-30%. The rank order (at 1 mM) to potentiate the 5-HT-induced current was: 5-OHi approximately 5-aminoindole approximately catechol > indole. The concentration-effect curve of 5-HT was shifted leftwards by 1 mM 5-OHi, resulting in a two-fold increase of the apparent affinity of 5-HT from 1.4 microM to 0.7 microM, without affecting the Hill coefficient. The time constant of reversal of activation of the 5-HT-induced ion current upon washout of the agonist was delayed by 1 mM 5-OHi from 4.0 sec to 12.8 sec. 5-HT3 receptor-gated single channel events in cell-attached patches in the presence and absence of 1 mM 5-OHi were indistinguishable, apart from a slight increase in the event frequency. The results suggest that 5-OHi and analogues potentiate the 5-HT3 receptor-mediated ion current by delaying agonist dissociation and thereby increase the probability of channel opening. From the increased apparent affinity of 5-HT and the non-surmountability of the potentiating effect, it is concluded that 5-OHi and analogues are allosteric modulators of 5-HT3 receptors. PMID- 9225291 TI - 5-HT3 receptors in outside-out patches of N1E-115 neuroblastoma cells: basic properties and effects of pentobarbital. AB - A fast solution exchange system (Dilger and Brett, 1990; Biophysics Journal 57: 723-731) with an exchange rate < 1 msec was used to study 5-HT3 (5-HT; 5 hydroxytryptamine) receptor-mediated currents in superfused outside-out patches of N1E-115 mouse neuroblastoma cells. At negative membrane potentials, 5-HT induced inward currents in a concentration-dependent manner (IC50 = 3.8 microM, Hill coefficient = 1.8). The mean peak current at a near-maximally effective 5-HT concentration of 30 microM was 20.6 pA. The 5-HT3 receptor antagonist ondansetron (0.3 nM) reversibly inhibited the 5-HT (30 microM) signal by approximately 50%. The currents induced during application of 30 microM 5-HT for 2 sec were characterized by inward rectification, a monophasic onset (tau ON = 37.5 msec) and, after reaching a peak, a monophasic decay (desensitization; tau OFF = 391 msec). Onset and decay were slower at lower 5-HT concentrations. The recovery of fully desensitized patches required a washout period of 45 sec. Pentobarbital inhibited 5-HT-induced (30 microM) currents in a concentration-dependent manner. The maximally obtainable inhibition with a given pentobarbital concentration was reached already when it was exclusively coapplied with 5-HT (IC50 = 135 microM. Hill coefficient = -0.7), since additional preexposure for at least 45 sec did not alter the concentration-response curve of pentobarbital. In conclusion, outside-out patches of N1E-115 cells are suitable to study the kinetic properties of 5-HT3 receptor channels. The results obtained in this model with pentobarbital are compatible with the suggestion that the inhibitory action of pentobarbital on 5-HT3 receptors is dependent on the agonist-activated (open) channel. PMID- 9225292 TI - Radioligand binding and photoaffinity labelling studies show a direct interaction of phenothiazines at 5-HT3 receptors. AB - The effects of a range of phenothiazines were examined on 5-hydroxytryptamine3 (5 HT3) receptors in membranes from NIE-115 neuroblastoma cells using radioligand binding. Chlorpromazine, fluphenazine, perphenazine, trifluoperazine and prochlorperazine inhibited specific binding of both the 5-HT3 receptor antagonist [3H]GR65630 and agonist [3H]meta-chlorophenylbiguanide (mCPBG), with Ki values ranging from 0.4 to 3.9 microM. The mode of action of chlorpromazine was further examined using photoaffinity labelling in the presence and absence of 5-HT. Saturation radioligand binding data with both [3H]GR65630 and [3H]mCPBG showed that photoaffinity labelling with chlorpromazine (1 microM) caused a decrease in the maximum number of binding sites observed (35% and 28% for agonist and antagonist, respectively). This decrease was not observed when the membranes were incubated in the presence of 5-HT. The results demonstrate a direct interaction of a range of phenothiazines at the 5-HT3 receptor binding site. PMID- 9225293 TI - [3H]RS 57639, a high affinity, selective 5-HT4 receptor partial agonist, specifically labels guinea-pig striatal and rat cloned (5-HT4S and 5-HT4L) receptors. AB - RS 57639, by being a partial agonist in rat esophagus but a competitive antagonist in guinea-pig ileum, is one of several ligands which operationally discriminate among 5-HT4 receptors in different tissues. The discovery of splice variants of the 5-HT4 receptor, 5-HT4S and 5-HT4L, raises the possibility that this functional heterogeneity among 5-HT4 receptors may be due to differences in the interaction of ligands with different isoforms of the receptor. To test this idea, the functional and binding interactions of RS 57639 with rat 5-HT4S and 5 HT4L receptors were characterized. RS 57639 stimulated adenylate cyclase in cells expressing 5-HT4S or 5-HT4L receptors with similar potency (pEC50 = 7.9 +/- 0.1 and 7.6 +/- 0.1) and efficacy (71 +/- 3 and 59 +/- 4% of 5-HT). [3H]RS 57639 also bound to 5-HT4S and 5-HT4L receptors with similar affinity (Kd = 0.09 +/- 0.01 and 0.11 +/- 0.01 nM) and specificity (SB204070 > GR113808 > SDZ 205557 > cisapride > renzapride > alpha me-5-HT > 5-CT). Therefore, the operational differences among 5-HT4 receptors, detected with RS 57639, are not explained by differences in the interaction of the ligand with 5-HT4S and 5-HT4L receptors. [3H]RS 57639 binding to guinea-pig striatal membranes was also characterized. [3H]RS 57639 bound with high affinity (Kd = 0.25 +/- 0.07 nM) and a specificity similar to that of the 5-HT4 receptor antagonist, [3H]GR113808. Therefore, while the mechanism by which RS 57639 operationally distinguishes among 5-HT4 receptors was not determined, [3H]RS 57639 was shown to specifically label native and cloned 5-HT4 receptors. As the first selective agonist radioligand to be described for this receptor, [3H]RS 57639 may prove useful in further studies of receptor coupling and ligand interactions. PMID- 9225294 TI - 5-HT4 receptors improve social olfactory memory in the rat. AB - Serotonin (5-HT) is involved in a large variety of physiological functions and it appears now that it could play a role in cognitive processes through the activation of 5-HT4 receptors. The present study was conducted to determine the effect of BIMU1, a mixed 5-HT4 agonist/5-HT3 antagonist on social olfactory recognition in rats, a behaviour test which has previously been shown to access short-term memory and to be sensitive to cholinergic drugs. This test is based on the investigation of an unfamiliar juvenile by an adult rat during two distinct 5 min presentations. At a 30-min delay after each presentation adults recognized the juvenile, whereas after a 2-hr delay all the adults had forgotten it. When administered intraperitoneally immediately after the first presentation, BIMU1 (10 mg/kg) enhanced short-term memory (i.e. recognition of the juvenile after a 2 hr delay). Ondansetron (10 and 100 micrograms/kg injected intraperitoneally), a 5 HT3 antagonist, had no significant effect on this form of memory. The effect of BIMU1 was antagonized by intraperitoneal injection of GR 125487, a very selective and potent 5-HT4 antagonist. The antagonistic effect was obtained at 1 and 10 mg/kg of GR 125487, but not at 0.1 mg/kg. It is certainly a specific effect on brain 5-HT4 receptors, since we determined a brain concentration of GR 125487 equal to 3.8 x 10(-7) M after the intraperitoneal injection of 10 mg/kg of this drug. This GR 125487 concentration is certainly sufficient to occupy all the 5 HT4 brain receptors (Kd = 10(-10) M) but not to occupy 5-HT3 receptors (Kd > 10( 6) M). The 5-HT4 specificity of the blockade by GR 125487 is further demonstrated by the fact that a 10-fold lower dose of GR 125487 (1 mg/kg) is also effective to inhibit the BIMU1 effect. PMID- 9225295 TI - The effects of novel, selective 5-hydroxytryptamine (5-HT)4 receptor ligands in rat spatial navigation. AB - Activation of central 5-hydroxytryptamine (5-HT4) receptors may enhance cognitive performance. In the present study, the effects of two novel, potent and selective 5-HT4 receptor agonists, RS 67333 (1-(4-amino-5-chloro-2-methoxyphenyl)-3-(1-n burtl-4-piperidinyl)- 1-propanone) and RS 67506 (1-(4-amino- 5-chloro-2 methoxyphenyl)-3-[1-[2-[(methylsulfonyl)amino]ethyl]-4- piperidinyl]-1 propanone), were studied in a rat model of spatial learning and memory; the Morris water maze. RS 67333 (0.1, 10 and 1000 micrograms/kg, intraperitoneally (i.p.)), a highly potent, selective and hydrophobic 5-HT4 receptor agonist, reversed the decrements in cognitive performance induced by atropine (30 mg/kg, i.p.). By contrast, no effect was seen to RS 67506 (0.1, 10 and 1000 micrograms/kg, i.p.), a hydrophilic 5-HT4 receptor agonist, of equivalent potency and selectivity to RS 67333. This differential effect may reflect the enhanced ability of RS 67333 to enter the CNS, with respect to RS 67506. The ameliorative actions of RS 67333 on cognitive dysfunction were abolished by prior treatment with a selective 5-HT4 receptor antagonist, RS 67532 [1-(4-amino-5-chloro-2-(3, 5 dimethoxy benzyloxyphenyl)-5-(1-piperidinyl)-1-pentanone; 1 mg/kg, i.p.]. When given alone, or in naive rats, RS 67532 (0.1, 10 and 1000 micrograms/kg, i.p.), was without effect. None of the compounds tested affected the swim speed at any of the doses used. In separate locomotor studies, RS 67532 reduced activity at 1 and 10 mg/kg, i.p., although no effect was seen with RS 67333 or RS 67506 (0.01 10 mg/kg, i.p.). These data suggest that RS 67333 reversed the cognitive deficit induced by atropine and support a role of 5-HT4 receptors in rat spatial learning and memory. PMID- 9225296 TI - BIMU1 increases associative memory in rats by activating 5-HT4 receptors. AB - Olfactory association learning was used to investigate the involvement of 5-HT4 receptors in learning and long-term memory. The behavioral role of the 5-HT4 receptors was studied by using BIMU1 (3-ethyl-2,3-dihydro-N-[endo-8-methyl-8 azabicyclo(3.2.1)oct-3-yl]-2-oxo -1 H-benzimidazole-1-carboxamide, hydrochloride (Boehringer Ingelheim, Italy); a mixed 5-HT4 agonist/5-HT3 antagonist, and GR125487 (1-[2-[methyl sulphonyl)-amino]ethyl]-4-piperidinyl-methyl 5-fluro-2 methoxy-1H-indole-3- carboxylate; Glaxo Group Research, Hertfordshire, U.K.), a specific 5-HT4 antagonist. The intraperitoneal injections of BIMU1 at 1, 5, and 10 mg/kg were followed by an substantial improvement (> 15% in percentage of correct responses at the dose of 10 mg/kg) in associative memory. Difficulty rapidly reversing behavioral responses to previously learned association, 1 month later indicated that the BIMU1 effect at 10 mg/kg was not transient, but correlated to long-term memory. The effects of BIMU1 are most likely to be mediated by 5-HT4 receptors since they were blocked by GR125487 at 10 mg/kg. These data suggest that activation of 5-HT4 receptors may modulate cognitive processes like learning and memory. PMID- 9225297 TI - Anxiolytic-like actions of the selective 5-HT4 receptor antagonists SB 204070A and SB 207266A in rats. AB - The highly selective 5-HT4 receptor antagonists, SB 204070A (0.001-0.1 mg/kg s.c., 30 min pretest) and SB 207266A (0.01, 1 and 10 mg/kg p.o., 1 hr pre-test), increased time spent in social interaction without affecting locomotor activity, in a rat 15 min social interaction test under high light, unfamiliar conditions. At 1 and 10 mg/kg s.c., SB 204070A was no longer active. These results are consistent with the profile expected of anxiolytic treatments in this procedure. In a rat 5 min elevated x-maze test, SB 204070A (0.01 and 1 mg/kg s.c., 30 min pre-test) significantly increased the percentage of time spent on the open arms. SB 204070A (0.01 mg/kg s.c.) and SB 207266A (1 mg/kg p.o., 1 hr pre-test) also increased percentage entries to the open arms. Neither compound affected locomotion at any dose tested in the procedure. The effects of both compounds in this procedure are also consistent with anxiolysis. Neither SB 204070A (0.1 or 1 mg/kg s.c., 30 min pre-test) nor SB 207266A (0.1 or 1 mg/kg p.o., 1 hr pre-test) affected either unpunished or punished responding, in a rat Geller-Seifter conflict model of anxiety. The maximal efficacy of both SB 204070A and SB 207266A in the rat social interaction test was similar to that of the benzodiazepine anxiolytic chlordiazepoxide (5 mg/kg s.c. or p.o.) used as a positive control, but was considerably less in the elevated x-maze procedure. The results suggest that 5-HT4 receptor antagonists may have modest anxiolytic-like actions in rats. PMID- 9225298 TI - Functional and radioligand binding characterization of rat 5-HT6 receptors stably expressed in HEK293 cells. AB - We have stably expressed the rat 5-HT6 receptor in HEK293 cells at a density of > 2 pmol/mg protein, as determined in equilibrium binding studies with [3H]-LSD and [3H]-5-HT and compared the affinity of a range of compounds in competition binding experiments with either [3H]-LSD or [3H]-5-HT as radioligand. A variety of tryptamine derivatives were tested and showed a significantly higher affinity when the 5-HT6 receptor was labelled with [3H]-5-HT, whereas ergoline compounds and several antagonists had higher affinities when [3H]-LSD was used as radioligand. Subsequently we examined the ability of LSD, 5-HT and a number of tryptamine derivatives to stimulate cAMP accumulation in order to determine their agonist potency and efficacy. We observed the following rank order of potency: LSD > omega-N-methyl-5-HT approximately bufotenine approximately 5- methoxytryptamine > 5-HT > 2-methyl-5-HT approximately 5-benzyloxytryptamine approximately tryptamine > 5-carboxamidotryptamine > > 5-HTQ. LSD, lisuride, 2 methyl-5-HT, tryptamine and 5-benzyloxytryptamine behaved as partial agonists relative to 5-HT. The rank order of potency of the tryptamine compounds correlated well with their affinities determined in binding assays. In addition, we have tested a number of antagonists in this system (rank order of potency: methiothepin, clozapine, mianserin and ritanserin). This characterization of the pharmacological properties of recombinant 5-HT6 receptor will facilitate the identification of 5-HT6 receptor-mediated responses in physiological systems. PMID- 9225299 TI - Modulation of IH by 5-HT in neonatal rat motoneurones in vitro: mediation through a phosphorylation independent action of cAMP. AB - The depolarization of adult and neonatal rat facial and spinal motoneurones by 5 hydroxytryptamine (5-HT) in part involves an enhancement of the hyperpolarization activated, inward-rectifier, IH. Under experimental conditions which promote this action, 5-HT evokes an inward current which can be mimicked by intracellularly applied adenosine 3',5'-cyclic monophosphate (cAMP) and potentiated by the cAMP specific phosphodiesterase inhibitor Ro 20-1724. In this study, we show that this action of 5-HT can be blocked by the adenylyl cyclase inhibitors 2'3' dideoxyadenosine (2',3'-DDA). 5'-adenylimidodiphosphate (AMP-PNP) and SQ-22536 (9 (tetrahydro-2-furyl)adenine), but not by external or internal application of the protein kinase inhibitors H-7, staurosporine and chelerythrine. The most recently cloned 5-HT receptor subtypes, 5-HT4, 5-HT6 and 5-HT7, can all stimulate adenylyl cyclase when activated. In the presence of internal GTP-gamma-S, 5-HT irreversibly enhanced IH. The 5-HT-induced inward current could be reversibly blocked by methysergide, but not by the 5-HT4 receptor antagonist GR-113808A, the 5-HT6 and 5-HT7 antagonist clozapine and the 5-HT1A antagonist WAY-100365. 5 Methoxytryptamine (5-MeOT) and 5-carboxamidotryptamine (5-CT) mimicked the action of 5-HT with a rank order of potency of 5-HT = 5MeOT > 5-CT. Surprisingly, 8 hydroxy-2-(di-N-propylamino)-tetralin (8-OH DPAT), a 5-HT1A and 5-HT7 agonist was inactive on facial motoneurones unlike its reported agonist action on spinal motoneurones. It is proposed that cAMP produced by 5-HT-mediated stimulation of adenylyl cyclase acts in a phosphorylation-independent manner, possibly directly, on the IH channel. The 5-HT receptor subtype mediating this response cannot be correlated with any of the classified 5-HT receptor subtypes that stimulate adenylyl cyclase. PMID- 9225300 TI - Comparative study in the rat of the actions of different types of stress on the release of 5-HT in raphe nuclei and forebrain areas. AB - The effects of several stress procedures on the release of 5-HT in the dorsal and median raphe nuclei (DRN and MRN, respectively) and in forebrain structures of the rat brain innervated by both nuclei have been studied using intracerebral microdialysis. Handling for 30 sec, a saline injection and forced swimming for 5 min elevated significantly the 5-HT output in the MRN. The 5-HT output in the DRN was also enhanced by a saline injection. With regard to the forebrain structure examined, handling and forced swimming increased dialysate 5-HT in the amygdala. The injection of saline induced a slight, but significant, elevation of 5-HT in the medial prefrontal cortex. In contrast, the outflow of 5-HT was significantly reduced in the ventral hippocampus and medial prefrontal cortex following forced swimming and this effect persisted well beyond the cessation of the swim session. These results indicate that the efflux of 5-HT in the MRN appears to respond to different forms of stress, whereas that in the DRN only increases after the injection of saline. The release of 5-HT in the forebrain structures is also dependent on the type of stress procedure and the region studied. PMID- 9225301 TI - 8-OH-DPAT-induced spontaneous tail-flicks in the rat are facilitated by the selective serotonin (5-HT)2C agonist, RO 60-0175: blockade of its actions by the novel 5-HT2C receptor antagonist SB 206,553. AB - The 5-HT1A receptor agonist, 8-OH-DPAT ((+/-)-8-dihydroxy-2-(di-n-propylamino) tetralin), (0.63 mg/kg, s.c.) elicited spontaneous tail-flicks (STFs) in rats. This response was potentiated by the selective 5-HT2C receptor agonist, RO 60 0175 ((S)-2-(6-chloro-5-fluoroindol-1-yl)-1-methylethylamine) fumarate) (0.16 mg/kg, s.c.), the action of which was abolished by the novel 5-HT2C antagonist, SB 206,553 (5 methyl-1-(3-pyridil-carbamoyl)-1,2,3,5-tetrahydropyrrolo[2,3 f]indole) (0.16 mg/kg, s.c.). These data show that 5-HT1A receptor-mediated STFs in rats are facilitated by activation of 5-HT2C receptors supporting the existence of functional interactions between these sites. PMID- 9225302 TI - Potassium channelopathies. AB - The molecular diversity of K(+)-selective channels far exceeds any other group of voltage- or ligand-gated channels, reflecting their early ancestral origin. This diversity is mirrored by the broad spectrum of physiological functions subserved by these proteins. Potassium channels modulate the resting potential and action potential duration of neurons, myocytes and endocrine cells and stabilize the membrane potential of excitable and nonexcitable cells. In addition to channel diversity, differential cellular expression of K+ channels determines the specific electrical responses to stimuli in a particular cell or tissue. This study reviews the recent genetic and physiological studies of congenital disorders caused by mutations in genes encoding K+ channels. These include the human disorders of episodic ataxia with myokymia, long QT syndrome and Bartter's syndrome, and weaver ataxia in mice. An understanding of the molecular basis of these diseases could facilitate the discovery and development of specific pharmacological therapies. PMID- 9225303 TI - Regulation of plasma aldosterone levels by metoclopramide: a reappraisal of its mechanism from dopaminergic antagonism to serotonergic agonism. AB - It has been thought, since the late 1970s, that dopamine exerts a tonic suppression of plasma aldosterone levels in human subjects. This action, however, had not been established directly using dopamine and dopamine mimetic drugs, which do not, in fact, affect the aldosterone levels. Rather, the conclusion was arrived at indirectly, based on the increase in aldosterone levels seen with dopamine receptor blockers; metoclopramide in particular, considered at the time of its discovery in the 1960s to be a new generation dopamine antagonist. However, metoclopramide is not a pure drug and in fact, shows intermediate affinity at certain serotonin receptor subtypes. Studies have been recently carried out in human subjects on the role of serotonergic transmission in mediating the metoclopramide as an aldosterone secretagogue effect. Here we briefly review this work and attempt to reassess the action of metoclopramide as an aldosterone secretagogue, from dopamine D2 antagonism to serotonin 5-HT4 partial agonism. PMID- 9225304 TI - Involvement of dopamine D2 receptors in the effect of cocaine on sexual behaviour and stretching-yawning of male rats. AB - The effect of cocaine (7.5, 15 and 30 mg/kg) administered in acute or subchronic mode, on the mating behaviour of sexually active male rats varied in a dose- and mode-dependent manner. Regardless of mode of treatment, 30 mg/kg markedly impaired the rats copulatory ability and impairment continued for a week after suspension of subchronic treatment. An acute dose of 15 mg/kg reduced intromission frequency, while in subchronic mode it also reduced ejaculation latency. Mount frequency was increased by 7.5 and 15 mg/kg, but only on first injection. In the case of sexually-naive male rats, acute administration of cocaine (3-30 mg/kg) stimulated penile erections at 7.5 mg/kg and motor hyperactivity at all doses. (-) Eticlopride (0.025 and 0.05 mg/kg), a DA D2 antagonist, counteracted cocaine-induced motor hyperactivity but not penile erection, which it enhanced. (-) Eticlopride at the same doses also antagonized cocaine potentiation of lisuride (0.2 mg/kg)-induced behavioural effects. When male rats treated with subchronic cocaine (15 mg/kg) were injected with the DA D2 agonist SND 919 (0.1 mg/kg), they displayed a more marked stretching-yawning behaviour than control animals receiving SND 919 at the same dose. The involvement of DA D2 receptors in cocaine-induced effects is suggested. PMID- 9225305 TI - [3H]raclopride binding to brain tissue from subjects with schizophrenia: methodological aspects. AB - The binding of [3H]raclopride to particulate membrane and frozen sections (with quantitative autoradiography) from the caudate-putamen, obtained at autopsy from schizophrenic and non-schizophrenic subjects, was measured. The affinity of [3H]raclopride to particulate membrane was significantly decreased in the schizophrenic compared to non-schizophrenic subjects. The density of [3H]raclopride binding to tissue from subjects with schizophrenia was increased, unchanged or decreased depending on the methodology used. Finally, there was an age-dependent decrease in [3H]raclopride binding in the frozen sections from the caudate-putamen of the non-schizophrenic subjects. This age-dependent decrease was not apparent using particulate membrane from schizophrenic or non schizophrenic subjects or tissue sections from the schizophrenic subjects. We conclude that the binding of [3H]raclopride is dependent on methodology and therefore data from in vitro and in vivo studies using this drug should be interpreted with caution. PMID- 9225306 TI - Evidence to suggest that agonist modulation of hyperlocomotion is via post synaptic dopamine D2 or D3 receptors. AB - It has been suggested that a sub-population of dopamine D3 receptors is located pre-synaptically and these serve as autoreceptors in dopamine projection areas such as the nucleus accumbens/ventral striatum. To study further the physiological role and synaptic location of the dopamine D3 receptor, we have investigated the in vivo effect of the D3/D2 receptor agonist quinelorane on amphetamine-induced hyperactivity and extracellular dopamine release from the nucleus accumbens of the conscious rat. Amphetamine increased dopamine release to 202 +/- 34% of pre-injection control values, but quinelorane at 2.5 micrograms/kg, a dose which effectively blocked amphetamine-induced hyperlocomotion, had no significant effect on amphetamine-induced dopamine release. These data suggest that hyperlocomotion is mediated via post-synaptic rather than pre-synaptic dopamine receptors. Since quinelorane has significant affinity for the dopamine D3 receptor, these effects may be via post-synaptic D3 receptors; however, D2 receptor effects cannot be disregarded. In summary, these data indicate that the quinelorane effect on amphetamine-stimulated hyperlocomotion is not mediated via D3 or D2 autoreceptors, but rather a population of receptors located post-synaptically, which appear to mediate the inhibition of rat locomotor activity. PMID- 9225307 TI - A comparative study of the effects of selective and non-selective 5-HT2 receptor subtype antagonists in rat and mouse models of anxiety. AB - Although there is some evidence that compounds acting at 5-HT2 receptors show anxiolytic activity, little is known about the specific involvement of the different 5-HT2 receptor subtypes in the modulation of anxiety-related responses. In the present study, the behavioural effects of mianserin, a non-selective 5-HT2 receptor antagonist, MDL 100,907, a selective 5-HT2A receptor antagonist, and SB 206553, a selective 5-HT2B/2C receptor antagonist, were investigated in two rat (the Vogel drinking conflict and the elevated plus-maze tests) and two mouse (i.e. the mouse defense test battery (MDTB) and the light/dark choice test) models of anxiety. Diazepam was used as a positive control. In the Vogel drinking test, mianserin (10 mg/kg) and SB 206553 (3-30 mg/kg), but not MDL 100,907, increased punished responding. Similarly, mianserin (1 mg/kg) and SB 206553 (3-10 mg/kg), but not MDL 100,907, increased entries into the open arms of the elevated plus-maze. These effects are consistent with anxiolytic-like actions of mianserin and SB 206553, although the magnitude of the effects of these two compounds was less than those of diazepam. In addition, in the MDTB, the 5-HT2 antagonists did not clearly affect the defensive reactions of mice exposed to a rat stimulus and they failed to reverse the avoidance of the illuminated box in the light/dark choice test. These results indicate a lack of anxiolytic-like action of the compounds in mice. These behavioural profiles suggest that blockade of the 5-HT2A receptor may not reduce anxiety and demonstrate that 5-HT2B and/or 5-HT2C receptor subtypes may be primarily involved in the anxiolytic-like effects of mianserin and SB 206553 in rats. PMID- 9225309 TI - Characterization of the cell death process induced by staurosporine in human neuroblastoma cell lines. AB - Staurosporine is a potent and non-specific inhibitor of protein kinases. There is also evidence of staurosporine being a potent inducer of apoptosis. In several human neuroblastoma cell lines (SH-SY5Y, NB69, IMR-5 and IMR-32) we have found 100 nM staurosporine to induce cell death in half the population (EC50). Electron microscopy of these cells, fluorescence microscopy after Hoechst-33258 staining of chromatin and agarose-electrophoresis of DNA, show two different types of cell death. SH-SY5Y and NB69 die by apoptosis and display all the characteristic features of it. IMR-5 and IMR-32 lack some of these features and a ladder pattern of DNA degradation is not found. Different morphological types of apoptosis have been described during the development of vertebrates; the possibility of finding a similar diversity in cell culture is suggested. On the other hand, staurosporine is a potent promoter of neurite outgrowth. In all the neuroblastoma cell lines we have tested, neurite-promoting and cell death-inducing staurosporine concentrations mostly overlap. This fact has not been reported before, probably because of an early versus late timing of these two different phenomena. The neuritogenic effect has prompted the suggestion that staurosporine could be a prototype of drugs for neurodegenerative diseases; the present study raises several concerns about such a proposal. PMID- 9225308 TI - Carrier-mediated serotonin release induced by d-fenfluramine: studies with human neuroblastoma cells transfected with a rat serotonin transporter. AB - The NMB human neuronal cell line, transfected with a newly prepared plasmid expressing rat serotonin transporter (NMB-rSERT), shows specific [3H]5-HT uptake which is blocked by citalopram and fenfluramine (F) stereoisomers with IC50 values (1 nM. 0.5 microM (dF) and and 5 microM (IF), respectively) which are similar to those found in rat brain synaptosomes. d-Fenfluramine (0.5 and 10 microM) also stimulates tritium release from NMB-rSERT cells preloaded with [3H-] 5-HT. The d-fenfluramine-induced [3H-]5-HT release is blocked by 0.3 microM citalopram and is dependent on the density of SERT expressed per cell, but is not affected by removal of Ca++ ions from the incubation medium. Manipulation of the Na+ gradient across the plasma membrane (replacing 60 mM NaCl with an equimolar concentration of KCl or choline) also induced [3H-]5-HT release from NMB-rSERT cells, which was inhibited by 0.3 microM citalopram. These results, together with the finding that NMB-rSERT cells preloaded with 500 nM unlabelled 5-HT take up [3H-]d-fenfluramine, make NMB-rSERT cells a valuable tool for studying the transporter-mediated exchange release induced by amphetamine derivatives. PMID- 9225310 TI - Modulation of long-term potentiation in CA1 region of mouse hippocampal brain slices by GABAA receptor benzodiazepine site ligands. AB - Enhancement of GABAA receptor function with benzodiazepine (BZ) site agonists can disrupt memory formation and hippocampal synaptic plasticity. To investigate this further the effects of the agonist, flunitrazepam, were contrasted with that of the inverse agonist, methyl-6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM), on NMDA-dependent LTP induction in the CA1 region of mouse hippocampus. Under control conditions, a priming stimulus (10 stimuli at 100 Hz) potentiated e.p.s.p. slopes by 198%, and subsequent burst stimuli (4 x 10 events at 100 Hz every 20 sec) by 306%. This potentiation was blocked by the non-competitive NMDA receptor antagonist MK-801 and the glycine site antagonist L-701,324. Flunitrazepam (1 microM) alone caused a slight but significant reduction in e.p.s.p.s to 83% of control, suppressed LTP induced by priming stimuli (133%) and burst stimuli (188%), but not that induced by sustained high-frequency stimulation (2 x 100 events at 100 Hz, 20 sec apart). The suppression of LTP induction by flunitrazepam was blocked by the benzodiazepine site antagonist flumazenil. In contrast, the inverse agonist DMCM (100 nM) potentiated LTP formed by both priming (to 283%) and burst stimuli (to 477%). This was associated with an enhancement of paired pulse facilitation during the induction phase and the subsequent appearance of paroxysmal burst discharges. Therefore, in addition to improvements in learning and memory as a result of improved vigilance, benzodiazepine inverse agonists can have direct effects on synaptic processes thought to contribute to memory formation. PMID- 9225311 TI - Metabotropic glutamate receptor mediated long-term depression in developing hippocampus. AB - The effects of bath application of the metabotropic glutamate receptor (mGluR) agonist 1S,3R-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD, 10 microM) were studied at the Schaffer collateral-CA1 synapse in hippocampal slices from rats of 8-33 days postnatal age. In immature animals (8-12 days) ACPD induced a biphasic response characterized by an acute decrease in field EPSP slope (approximately 50 60% of baseline) in the presence of the agonist, followed by long-term depression (LTD, approximately 75-80% of baseline) after washout. In animals older than 20 days, ACPD induced a slow onset potentiation or minimal change. Both the acute depression and LTD were blocked by the mGluR antagonist alpha-methyl-4 carboxyphenyl glycine (MCPG). ACPD-induced LTD was blocked by the N-methyl-D aspartate receptor (NMDAR) antagonists D(-)-2-amino-5 phosphopentanoic acid (AP5) and dizocilpine maleate (MK-801), and by ethanol. Glutamic pyruvic transaminase, an enzyme that selectively metabolizes endogenous extracellular glutamate, also blocked LTD suggesting that the requisite NMDA currents were tonically activated by extracellular rather than synaptically released glutamate. ACPD-induced LTD was blocked by staurosporine, indicating a requirement for serinethreonine kinase activation, and was unaffected by the L-type voltage sensitive calcium channel blocker nitrendipine and the A1 adenosine receptor antagonist 8-cyclopentyl-1,3 dimethylxanthine (CPT). Because mGluR-mediated LTD was observed only in immature CA1, mGluRs may play a role in hippocampal development, perhaps by contributing to synapse pruning in a temporally restricted fashion. PMID- 9225312 TI - Activation of group III metabotropic glutamate receptors depresses glutamatergic transmission at corticostriatal synapse. AB - Intracellular recordings were performed from a rat corticostriatal slice preparation in order to characterize the effects of group III metabotropic glutamate receptor (mGluR) agonists on excitatory transmission at corticostriatal synapses. The amplitude of excitatory postsynaptic potentials (EPSPs), evoked by cortical stimulation, was significantly decreased by agonists acting at group III metabotropic glutamate receptors. Both L-2-amino-4-phosphonobutanoate (L-AP4) and L-serine-O-phosphate (L-SOP) were effective in reducing the amplitude of cortically evoked EPSPs, in a dose-dependent manner. The EC50 value for the effect of L-SOP and L-AP4 was 0.89 microM and 9.95 microM, respectively. Both L AP4 and L-SOP had negligible effects on the intrinsic membrane properties of the recorded neurons and did not alter the postsynaptic response to focal application of glutamate, suggesting a presynaptic site of action. The presynaptic inhibition of both L-SOP and L-AP4 was fully antagonized by 250 microM (s)-2-methyl-2-amino 4-phosphonobutanoate (MAP4), whilst it was unaffected by 500 microM RS-methyl-4 carboxyphenylglycine (MCPG). Conversely, the presynaptic inhibitory effect on the EPSP amplitude exerted by 10 microM 1S,3R-1-aminocyclopentane-1,3-dicarboxylic acid (1S,3R-ACPD) was antagonized by 500 microM MCPG, whilst it was not blocked by 250 microM MAP4. Finally, the reduction of the EPSP amplitude produced by a saturating dose of L-SOP was further increased by 10 microM 1S,3R-ACPD, suggesting an additive effect of these compounds. The present results are consistent with the idea that group III mGluRs exert a presynaptic inhibitory modulation of the excitatory glutamatergic transmission at corticostriatal synapses. PMID- 9225313 TI - Desensitization of kainate receptors by kainate, glutamate and diastereomers of 4 methylglutamate. AB - The potencies of kainate, glutamate and diastereomers of 4-methylglutamate were determined for activation and steady-state desensitization of GluR6 and dorsal root ganglion-type kainate receptors using whole-cell voltage clamp. In HEK293 cells expressing GluR6, all four diastereomers induced desensitizing inward currents at relatively high concentrations (> 50 microM), however, the 2S,4R diastereomer (2S,4R-4MG; SYM 2081) was approximately 100-fold more potent than the other three. The EC50 for receptor activation by 2S,4R-4MG (1.0 microM) was similar to that for kainic acid (1.8 microM), but 2S,4R-4MG was significantly more potent than kainate, glutamate or the other diastereomers of 4 methylglutamate at producing steady-state desensitization of GluR6 receptors. IC50s for desensitization quantified using a fixed concentration of kainate as a test agonist were 7.6, 31 and 667 nM for 2S,4R-4MG, kainate and glutamate, respectively. In addition, 2S,4R-4MG fully desensitized native kainate receptors (of the GluR5 subtype) in dorsal root ganglion neurons with an IC50 of 11 nM, compared to 3.4 microM for glutamate. For GluR6, recovery from desensitization displayed a similar time course for kainate and glutamate (tau = 3-4 s) but was roughly 20-fold slower for 2S,4R-4MG, which suggests that the rate of recovery is not entirely dependent on the affinity of ligand for the desensitized receptor. Following exposure to concanavalin A, application of kainate, glutamate and 2S,4R 4MG evoked very similar maximal currents that showed little or no desensitization. Lectin pretreatment produced a leftward shift in the concentration-response relationship for 2S,4R-4MG with an 11-fold reduction in the EC50; however, no significant change in the EC50 for kainate was observed. The characteristic of 2S,4R-4MG to potently and completely desensitize both recombinant GluR6 receptors and native receptors on dorsal root ganglion neurons suggests that this compound will be useful to study selective blockade of these receptors in the nervous system. PMID- 9225314 TI - Pharmacologically distinct presynaptic calcium channels in cerebellar excitatory and inhibitory synapses. AB - We have used whole-cell patch clamp recordings and pharmacological blockers of Ca channels to compare the pharmacology of Ca channels that mediate synaptic transmission at the three types of synapses innervating Purkinje cells in rat cerebellar slices. Both parallel fiber and climbing fiber excitatory synapses were sensitive to the P-type Ca channel blocker, omega-AgaIVA and the P/Q/N-type channel blocker, omega-conotoxin MVIIC. Transmission at inhibitory interneuronal synapses was not suppressed by these toxins, or by the N-type (omega-conotoxins GVIA and MVIIA) or L-type (nimodipine) channel blockers. Inhibitory transmission could be inhibited by Ni2+ and amiloride, but only at concentrations (IC50 approximately 300 microM) that affect other types of Ca channels. These results indicate that excitatory and inhibitory presynaptic terminals of the cerebellar cortex possess different types of voltage-gated Ca channels. The excitatory terminals contain P-type, Q-type and N-type Ca channels, with P-type channels playing the most prominent role. The inhibitory terminals possess quite different type(s) of Ca channel. The heterogeneous distribution of Ca channel types should impart unique properties to transmitter release from the excitatory and inhibitory terminals. PMID- 9225315 TI - Construction of a standard reference for PET studies of methionine accumulation using a computerised brain atlas. AB - Positron emission tomography (PET) is valuable for assessing the biochemistry and physiology of the human brain. A computerised brain atlas has been developed which allows demonstration of anatomical regions of PET images and manipulation of these images into a standardised anatomical space. Once the images are in this standardised three-dimensional space it is possible to make comparisons between individuals and groups of individuals. We describe the use of this atlas in the generation of a set of mean reference images using methionine PET images of normal volunteers. PMID- 9225317 TI - Intravenous angiography in brain death: report of 140 patients. AB - We present our experience and discuss the value of cerebral intravenous digital subtraction angiography (IV DSA) in the diagnosis of brain death. A total of 140 patients presenting with clinical signs of brain death were studied by IV DSA. According to the angiographic appearance of the vertebrobasilar system, the patients were divided into four groups. Cessation of blood flow within the internal carotid arteries and their branches was consistently found. Attention is focused on 9 patients with persistent blood flow within the posterior fossa. In sedated patients in whom EEG and evoked brain-stem responses are non-diagnostic, or in order to shorten the observation time, transcranial Doppler should be performed to determine the appropriate moment for IV DSA, which is a reliable method of confirming brain death. PMID- 9225316 TI - MRI of pituitary adenomas in acromegaly. AB - Adenomas causing acromegaly represent at least a quarter of pituitary adenomas. We studied 12 patients presenting with active acromegaly due to a pituitary adenoma with a 1.5 T superconductive MRI unit. All had T1-weighted sagittal and coronal sections before and after Gd-DTPA; six had coronal T2-weighted images. Surgical correlation was obtained in seven patients. Histologically, there were eight growth hormone (GH)-secreting and three mixed [GH and prolactin (PRL) secreting] adenomas, and one secreting GH, PRL and follicle-stimulating hormone. Macroadenomas (10) were more frequent than microadenomas (2). No correlation was found between serum GH and tumour size. There were nine adenomas in the lateral part of the pituitary gland; seven showed lateral or infrasellar invasion. Homogeneous, isointense signal on T1- and T2-weighted images was observed in six cases. Heterogeneous adenomas had cystic or necrotic components. PMID- 9225318 TI - MRI of acute cerebral infarction: a comparison of FLAIR and T2-weighted fast spin echo imaging. AB - Fluid-attenuated inversion-recovery (FLAIR) sequences have been reported to provide high sensitivity to a wide range of central nervous system diseases. To our knowledge, however, FLAIR sequences have not been used to study patients with acute cerebral infarcts. We evaluated the usefulness of FLAIR sequences in this context. FLAIR sequences were acquired on a 0.5 T superconducting unit within 8 h of the onset in 19 patients (aged 26-80 years) with a total of 23 ischaemic lesions. The images were reviewed retrospectively by three neuroradiologists, and the FLAIR images were compared with T2-weighted fast spin-echo images. All but one of the ischaemic lesions involving grey matter was clearly demonstrated on FLAIR images as increased signal intensity in cortical or central grey matter. FLAIR images were particularly useful for detecting the hyperacute cortical infarcts within 3 h of onset, which were not readily detected on the spin-echo images. In 9 of 11 patients with complete proximal occlusion, the distal portion of the cerebral artery was visible as an area of high signal intensity on FLAIR images. PMID- 9225319 TI - Isolated inferior sagittal sinus thrombosis: a case report. AB - We present a case of isolated inferior sagittal sinus thrombosis shown on CT, MRI and angiography. This condition has not, to our knowledge, been described previously. PMID- 9225320 TI - Recurrent cerebral venous infarcts and superior vena cava obstruction: case report. AB - We report a patient with repeated venous infarcts in the occipital lobe and occlusion of the superior vena cava. The pathogenetic relationships between the superior vena cava occlusion and the brain infarcts are discussed. High pressure in the superior venous territory and incomplete patency of the transverse sinus are probably responsible for the venous infarcts. PMID- 9225321 TI - Magnetic resonance ventriculography with gadolinium DTPA: report of two cases. AB - We report intrathecal use of gadolinium DTPA for MRI of the cerebrospinal fluid (CSF). In two patients with leptomeningeal carcinomatosis, we injected 0.01 mmol gadolinium DTPA into the lateral ventricle via an Ommaya device. Coronal T1 weighted images of the head were obtained at 0.2 T prior to and after injection. There was pronounced enhancement of CSF close to the injection site, allowing good delineation of CSF and surrounding brain tissue. No side effects occurred. MRI with intrathecal administration of highly diluted gadolinium DTPA may be a promising alternative to conventional investigation of CSF-filled cavities using iodinated X-ray contrast media or radionuclides. PMID- 9225322 TI - MRI and CT in an autosomal-dominant, adult-onset leukodystrophy. AB - We report MRI findings in a family with an autosomal-dominant, adult-onset neurological disorder. The clinical picture, the white matter changes detected on MRI and the absence of any laboratory abnormality suggested the diagnosis of leukodystrophy with an unknown biochemical defect. Autosomal-dominant inheritance is extremely rare in this kind of disease, and most reported families have not undergone MRI. We performed MRI and clinical examination of 17 members of our family; 9 affected subjects, at different stages of the disease, were detected. The most characteristic MRI findings were initially symmetrical areas of signal change in the white matter of the trigonal region; demyelination extending thereafter to the frontal and parietal regions, partially involving subcortical white matter; the temporal lobe and optic radiations were less involved; the internal capsule and corpus callosum were involved later, in a dorsoventral direction; patchy demyelination was evident in the late stages in the brain stem; the cerebellum was spared even in the latest stages of the disease. While pathological examination is essential to characterise and classify these kinds of diseases, MRI can make substantial contributions to understanding their natural history, and to detect early signs of the disease. PMID- 9225323 TI - Serial imaging in MELAS. AB - We report two patients with fatal mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). Single-photon emission computed tomography (SPECT) with 123I-N-isopropyl-p-iodoamphetamine was more sensitive to the lesions than CT or MRI. SPECT showed focal hyperperfusion before or during the stroke and diffuse hypoperfusion of the brain, sparing the basal ganglia in the terminal stages. These findings support the theory that metabolic disturbance in the brain causes the "stroke" in MELAS. PMID- 9225324 TI - Disseminated intracerebral alveolar echinococcosis: CT and MRI. AB - Cerebral alveolar echinococcosis is rare and has a poor prognosis. We report an unusual case presenting with disseminated intracranial lesions secondary to primary hepatic infection. PMID- 9225326 TI - MRI in carcinomatous encephalitis. AB - We report a rare case of miliary brain metastases presenting with symptoms similar to encephalitis ("carcinomatous encephalitis"). Contrast-enhanced MRI demonstrated miliary metastases more distinctly than other imaging methods and reproduced the pathological features. PMID- 9225325 TI - Granulomatous amebic encephalitis caused by acanthamoeba. AB - Infections arising from free-living amebae are rare. They generally cause recognizable disease only in chronically ill, debilitated patients who are immune suppressed. Only about 70 cases of granulomatous amebic encephalitis have been reported. We present an unusual case of granulomatous encephalitis in a 35-year old man. Neurologic examination and laboratory tests were inconclusive. CT demonstrated bilateral low-density areas with mild mass effect in the cortex and subcortical white matter, which showed increased signal on T2-weighted MRI. Craniotomy and brain biopsy revealed granulomatous encephalitis with acanthamoeba organisms. Though non-specific, imaging can support the diagnosis of amebic encephalitis and direct biopsy. PMID- 9225328 TI - The MRI signs of spinal arachnoid diverticula. AB - Our goal was to find MRI signs of use for identifying a spinal arachnoid diverticulum. Three cases of spinal arachnoid diverticula, one extradural and two intradural, were examined on a 1.5 T imager. There was obvious mass effect on the adjacent structures in one case and increased signal intensity in the diverticulum on proton density- and T2-weighted images in two cases. Signal changes due to turbulent movement of the spinal fluid inside the diverticula were seen in all cases on sagittal fast spin-echo (FSE) proton density- and T2 weighted images; it was difficult to tell whether these signal changes imply a communication or are simply FSE artefacts. On contrast-enhanced studies, all cases showed partial enhancement inside the diverticula. There thus are four signs of diverticula: mass effect, the increased signal, signal void sign and partial enhancement; the last of these, the most reliable, has never been reported before. PMID- 9225327 TI - Magnetisation transfer ratio measurement in the cervical spinal cord: a preliminary study in multiple sclerosis. AB - MRI readily detects the lesions of multiple sclerosis (MS) in the brain and spinal cord. Conventional MRI sequences do not, however, permit distinction between the various pathological characteristics (oedema, demyelination, axonal loss and gliosis) of lesions in MS. Magnetisation transfer (MT) imaging may be more specific in distinguishing the pathologies responsible for disability in MS, namely demyelination and axonal loss, and therefore may have a potential role in monitoring treatment. We have applied MT imaging to the cervical spinal cord to see if it is feasible to measure MT ratios (MTR) in this region where pathological changes may result in considerable disability. We studied 12 patients with MS and 12 age- and sex-matched normal controls using a sagittal T2 weighted fast spin-echo sequence with and without an MT pulse. The median value for cervical cord mean MTR measurement in normal controls was 19.30% units (interquartile range 19.05-19.55), whereas values were significantly lower in MS patients (median = 17.95% units, interquartile range 17.25-19.00, P = 0.0004). There was a low intrarater variability for repeated mean MTR measurements. We conclude that it is possible to measure MTR in the cervical spinal cord, that a significant reduction occurs in patients with MS, and that there may be a role for this measure in future MS treatment trials. PMID- 9225329 TI - Extradural spinal meningioma: MRI. AB - We report a case of extradural spinal meningioma with pathologically proven features of malignant transformation. The MRI findings of extradural spinal meningioma and differences in the findings from intradural meningiomas are discussed. PMID- 9225330 TI - MRI of active otosclerosis. AB - Our aim was to determine whether MRI reliably shows pathology in patients with active otosclerosis (otospongiosis). We studied five patients with clinical and audiometric signs of this disorder and positive findings on high-resolution CT and tympanocochlear scintigraphy. Contrast enhancement of otospongiotic lesions was found in all affected ears, and could be topographically related to demineralised otospongiotic foci on CT. In lesions in the lateral wall of the labyrinth MRI sometimes showed the pathology better than CT, where partial-volume effects could be troublesome. PMID- 9225331 TI - Hypothalamic neuronal histamine: implications of its homeostatic control of energy metabolism. AB - In a series of studies on histaminergic functions in the hypothalamus, probes to manipulate activities of histaminergic neuron systems were applied to assess its physiologic and pathophysiologic implications using non-obese normal and Zucker obese rats, an animal model of genetic obesity. Food intake is suppressed by either activation of H1-receptor or inhibition of the H3-receptor in the ventromedial hypothalamus (VMH) or the paraventricular nucleus, each of which is involved in satiety regulation. Histamine neurons in the mesencephalic trigeminal sensory nucleus modulate masticatory functions, particularly eating speed through the mesencephalic trigeminal motor nucleus, and activation of the histamine neurons in the VMH suppress intake volume of feeding at meals. Energy deficiency in the brain, i.e., intraneuronal glucoprivation, activates neuronal histamine in the hypothalamus. Such low energy intake in turn accelerates glycogenolysis in the astrocytes to prevent the brain from energy deficit. Thus, both mastication and low energy intake act as afferent signals for activation of histaminergic nerve systems in the hypothalamus and result in enhancement of satiation. There is a rationale for efficacy of a very-low-calorie conventional Japanese diet as a therapeutic tool for weight reduction. Feeding circadian rhythm is modulated by manipulation of hypothalamic histamine neurons. Hypothalamic histamine neurons are activated by an increase in ambient temperature. Hypothalamic neuronal histamine controls adaptive behavior including a decrease in food intake and ambulation, and an increase in water intake to maintain body temperature to be normally constant. In addition, interleukin-1 beta, an endogenous pyrogen, enhanced turnover of neuronal histamine through prostaglandin E2 in the brain. Taken together, the histamine neuron system in the hypothalamus is essential for maintenance of thermoregulation through the direct and indirect control of adaptive behavior. Behavioral and metabolic abnormalities of obese Zucker rats including hyperphagia, disruption of feeding circadian rhythm, hyperlipidemia, hyperinsulinemia, and disturbance of thermoregulation are essentially derived from a defect in hypothalamic neuronal histamine. Abnormalities produced by depletion of neuronal histamine from the hypothalamus in normal rats mimic those of obese Zuckers. Grafting the lean Zucker fetal hypothalamus into the obese Zucker pups attenuates those abnormalities. These findings indicate that histamine nerve systems in the brain play a crucial role in maintaining homeostatic energy balance. PMID- 9225332 TI - Evaluation of bioelectrical impedance for prospective nutritional assessment in cystic fibrosis. AB - We have compared the use of bioelectrical impedance analysis (BIA) with anthropometry for the prediction of changes in total body potassium (TBK) in a group (n = 31) of children with cystic fibrosis. Linear regression analysis showed that TBK was highly correlated (r > 0.93) with height2/impedance, weight, height, and fat-free mass (FFM) estimated from skin-fold measurements. Changes in TBK were also correlated, but less well, with changes in height2/impedance, weight, height, and FFM (r = 0.69, 0.59, 0.44, and 0.40, respectively). The children were divided into two groups: those who had normal accretion of TBK (> 5%/y) and those who had suboptimal accretion of TBK (< 5%/y). Analysis of variance showed that the significant difference in the change in TBK between the groups was detectable by concomitant changes in impedance and weight but not by changes in height, FFM, or weight and height Z scores. The results of this study suggest that serial BIA measures may be useful as a predictor of progressive undernutrition and poor growth in children with cystic fibrosis. PMID- 9225333 TI - Catabolism of lipoprotein-X induced by infusion of 10% fat emulsion. AB - The clinical significance of lipoprotein-X (Lp-X) induced by intravenous infusion of 10% fat emulsion was assessed, with special reference to atherogenesis, by in vitro experiment using purified Lp-X from the sera of patients receiving Intralipid 10%. Lp-X appeared after long-term intravenous infusion of 10% fat emulsion in the patients with intestinal fistula due to the anastomotic leakage. To clarify the role of Lp-X in terms of atherogenicity, the cholesterol metabolism of Lp-X in macrophages as scavenger cells and in hepatocytes as parenchymal cells was studied. When [3H]cholesterol-labeled Lp-X or oxidized low density lipoprotein (o-LDL) was incubated with J-774 macrophages, the incorporation of Lp-X into macrophages was negligible compared with o-LDL. When Lp-X or high-density lipoprotein (HDL) was incubated with J-774 macrophages laden with [3H]cholesterol, the release of cholesterol from macrophages was enhanced by Lp-X as well as HDL. When [3H]cholesterol-labeled Lp-X LDL or HDL was incubated with the human hepatoma cell line of Hep G2 cells, the incorporation of Lp-X into Hep G2 cells was less than that of LDL, but similar to that of HDL. From these findings, it is suggested that the catabolism of Lp-X cholesterol generated with intravenous 10% fat emulsion was mediated by hepatocytes rather than by macrophages, indicating that the hyperlipidemia due to increased Lp-X may not be atherogenic. PMID- 9225334 TI - Dietary supplementation at home improves the regain of lean body mass after surgery. AB - Little is known about nutritional intake after discharge though it takes months to regain preoperative weight after gastrointestinal surgery. We studied whether a 4-mo intervention with dietary advice and protein-rich supplements would increase nutritional intake and gain in lean body mass (LBM) in patients who had undergone gastrointestinal surgery. Patients admitted for gastrointestinal surgery were randomized at discharge to serve as control patients (n = 47) or to receive intervention (n = 40). One month after discharge, the control patients had a nutritional intake (3-d diet record) comparable with the intake of the general population that did not increase further. During the 4 m, the intervention patients had an increased intake of protein (+22%) and energy (+16%), and an enhanced gain of LBM after 2 mo (control 0.8 kg versus intervention 2.1 kg; P = 0.009). After the 4-mo intervention, both LBM and fat were gained (control 1.7 kg LBM and 0.2 kg fat versus intervention 3.1 kg LBM and 1.5 kg fat; LBM: P = 0.029 and fat: P = 0.056). At discharge patients should increase protein intake to 1.5 g.kg-1.d-1 for 2 mo, e.g., by taking protein-rich liquid supplements. PMID- 9225335 TI - Contrasting effects of identical nutrients given parenterally or enterally after 70% hepatectomy: bacterial translocation. AB - High mortality occurs in rats with 70% hepatectomy fed intravenous (IV) total parenteral nutrition (TPN; 13.9% glucose, 4.17% amino acids, 1.46% fat, electrolytes, trace minerals, and vitamins providing 216 kcal.kg-1.d-1) but not when the identical nutrients are given at the same rate enterally (gastrostomy). We hypothesized that a difference in bacterial translocation (BT) was a contributing factor to this phenomenon. Forty-five male Sprague-Dawley rats (300 360 g) were divided into five groups and underwent the following: control (no operation), sham (intraperitoneal [IP] pentobarbital anesthesia, central venous and gastrostomy catheters, laparotomy, sham hepatectomy), standard oral feeding (SOF), TPN (IV nutrients), and total enteral nutrition (TEN; gastrostomy). The SOF, TPN, and TEN groups had IP pentobarbital anesthesia, central venous and gastrostomy catheters, and 70% hepatectomy. Postoperatively, control and SOF (both catheters plugged) rats ate a commercial rat chow and drank tap water ad libitum pre- and postoperatively. The sham, TPN, and TEN groups were given the identical infusate composition as above, but the nutrient concentrations were cut in half (110 kcal/kg) and three-quarters (165 kcal/kg) on postoperative days 1 and 2, respectively. At the end of postoperative day 2, all rats were euthanized. BT to mesenteric lymph nodes (MLNs), liver, spleen, and lungs was significantly higher in the TPN rats compared with all other groups, except that BT to the MLNs was similar in the TPN and TEN groups. Bacteremia was found only in the TPN rats. BT in TPN rats with 70% hepatectomy was significantly greater 48 h after operation than in those fed the identical nutrients enterally at the same rate; this correlates with the previously reported significantly greater mortality in rats with 70% hepatectomy receiving TPN. PMID- 9225336 TI - Asymmetry of the total body water prediction bias using the impedance index. AB - Our purpose was to prove on a geometric basis that the bias of total body water (TBW) prediction equations based on the impedance index is far greater in fluid overloading than in dehydration. We used formal evaluation of conventional bioimpedance regression equations in both normal and abnormal body fluid status. We plotted the hyperboloid function generated from a standard prediction equation for the TBW over the resistance-reactance (RXc) plane containing the bivariate tolerance intervals (ellipses) of the healthy population. The equation estimated 35 L TBW for the average man (both sexes) of 170 cm height. Leaving the center of the tolerance ellipses, over which the function was relatively flat, the predicted TBW rapidly increased to absurd values for the shorter vectors, indicating fluid overloading (e.g., > 100 L for R < 170 ohm). Migration of the longer impedance vectors beyond the upper pole of 95% tolerance ellipse, which is in the dehydration region, produced less biased estimates of TBW (e.g., < 22 L for the extreme R values > 850 ohm). Different formulas produced TBW prediction bias of the same order. Due to the hyperbolic shape, functions of the impedance index are critically dependent on the region of the RXc plane where they are calculated and they produce misleading results in patients with fluid overload. PMID- 9225337 TI - Early enteral nutrition in gastrointestinal surgery: a pilot study. AB - There is still some concern about the safety of early enteral nutrition (EN) to patients with recent anastomoses. A pilot trial was carried out on a prospective basis to evaluate the tolerance and clinical outcome of 56 patients who received early EN following gastrointestinal (GI) surgery. A continuous infusion of an elemental, peptide-based diet was administered using a nasointestinal feeding tube placed beyond the pylorus by the operating surgeon. Tube feeds were started at 6.07 +/- 4.99 h after surgery and advanced as tolerated to a rate of 60 mL/h on the third postoperative day. Patients received the diet either proximal or distal (in the case of gastrectomies) to their recent anastomosis. Forty-six patients met the inclusion criteria and were included in the analysis. EN was well tolerated with a low incidence of side effects (19.5%), nausea and vomiting being the most frequent. Oral feeding was started 2.89 +/- 1.28 d after surgery. There was one case of small bowel suture leakage, but no relationship to the tube feeding was established. Early EN appears to be a useful and safe therapeutic alternative for the postoperative management of patients undergoing GI surgery. It may contribute to faster recovery of bowel function and lead to a shorter hospital stay. Careful selection of patients is necessary in order to obtain the greatest benefit of early enteral feeding in this patient population. PMID- 9225338 TI - Metabolic and nutritional management of a patient with multiple enterocutaneous fistulas. PMID- 9225339 TI - Vitamin E and human health: rationale for determining recommended intake levels. AB - The recent literature provides strong evidence that vitamin E intakes much higher than the current recommendations can contribute to and/or improve human health. In fact, the available data indicate that at higher-than-current recommended intake levels, vitamin E affects several functions related to human health. For example, Vitamin E is required to protect polyunsaturated fatty acids (PUFAs) against auto-oxidation. The amount of vitamin E needed to protect PUFAs against oxidative damage is at least 0.4-0.8 mg vitamin E per gram PUFAs and may be in excess of 1.5 mg/g when diets contain higher-than-average levels of long-chain PUFAs. Based upon studies of vitamin E kinetics and metabolism, a daily vitamin E intake of 135-150 IU is suggested. Important functions such as protection against oxidative damage, immune response, and the propensity of platelets to adhere to the vessel wall are related to vitamin E intakes. Vitamin E intake of 40 IU/d was the least amount demonstrated to inhibit low-density lipoprotein oxidation; a dose-dependent effect was seen up to 800 IU/d. Vitamin E intakes of at least 60 IU/d enhanced immune responses and intakes of 200 IU-400 IU/d decreased platelet adhesion to the vessel wall. Based upon the effects of modulating these functions, it is hypothesized that vitamin E plays a pivotal role in the prevention of cardiovascular diseases. Indeed, many observational studies have reported vitamin E to reduce the risk of cardiovascular disease. Recent intervention studies corroborate these findings. Of equal importance, there is a solid body of literature that demonstrates that these and much higher vitamin E intakes are safe. PMID- 9225340 TI - Limitation of stunning in dog myocardium by nucleoside and nucleotide mixture, OG VI. AB - OG-VI is a solution composed of 30 mM inosine, 30 mM sodium 5'-guanylate, 30 mM cytidine, 22.5 mM uridine, and 7.5 mM thymidine, expecting to use for total parenteral nutrition. We examined the effect of OG-VI on myocardial contractile dysfunction during reperfusion after ischemia (myocardial stunning) in dogs. Pentobarbital-anesthetized dogs were subjected to 20-min left anterior descending coronary artery ligation followed by 30-min reperfusion. Saline, OG-VI or its constituents [inosine and sodium 5'-guanylate mixture (IG), and cytidine, uridine, and thymidine mixture (CUT)], or 5-amino-4-imidazole carboxamide riboside (AICAr) was infused at 0.1 mL.kg-1.min-1, starting 30 min before the ischemia. The contractile function was determined by ultrasonometry and assessed as % segment shortening (%SS). %SS was markedly decreased by ischemia, and returned toward pre-ischemic level after reperfusion, although the recovery was incomplete. The %SS was almost completely recovered by OG-VI and IG, and to a lesser extent by AICAr; CUT was ineffective. In the presence of 1 mg.kg-1 of 8 cyclopentyl-1,3-dipropylxanthine (DPCPX, a selective adenosine A1 receptor antagonist), cardioprotective effect of OG-VI on stunned myocardium was still observed. In conclusion, infusion of OG-VI improved myocardial contractile dysfunction in stunned myocardium. This effect was more potent than its constituents and AICAr. Adenosine A1 receptors are not involved in the mechanism. PMID- 9225342 TI - Dietary nucleotides: a conditional requirement. PMID- 9225341 TI - Dietary nucleotides may influence the humoral immunity in immunocompromised children. PMID- 9225343 TI - Experimental hepatectomy: the effect on the intestine and influence of various supplements. PMID- 9225344 TI - A new approach to estimate changes in total body water by bioelectrical impedance analysis. PMID- 9225346 TI - Colonic microflora: nutrition and health. PMID- 9225345 TI - Effect of home supplementation on lean body mass after surgery. PMID- 9225347 TI - Home parenteral nutrition in adults: new trends raise new questions. PMID- 9225348 TI - Healthy body weight standards. PMID- 9225349 TI - Glucocorticoids and energy expenditure: relevance to the regulation of energy balance in man. PMID- 9225350 TI - Vitamin A deficiency, child health, and survival. PMID- 9225351 TI - Artificial nutrition in cancer patients: an outlook from Europe. PMID- 9225352 TI - Welfare reform: unanticipated but inevitable consequences for health insurance coverage for the poor. PMID- 9225353 TI - The role of the lung in the metabolism of fat. 1930. PMID- 9225355 TI - Paroxysmal movement disorders. PMID- 9225354 TI - A pediatrician's view. The fun of the chase. PMID- 9225356 TI - Breath-holding spells. PMID- 9225357 TI - Complicated migraine syndromes and migraine variants. PMID- 9225358 TI - The diagnosis and management of syncope in children and adolescents. PMID- 9225359 TI - Parasomnias in children. PMID- 9225360 TI - How I became a pharmacologist. PMID- 9225361 TI - Comparative study on the effect of calcium channel blockers on basal and parathyroid hormone-induced bone resorption in vitro. AB - A number of clinical and experimental studies suggest that the effects of calcium channel blockers are not limited to the cardiovascular system but might also involve skeletal calcium metabolism due to the presence of L-type calcium channels in osteoblastic cells. We therefore investigated the influence of calcium channel blockers of the dihydropyridine type (nifedipine, amlodipine) as well as of the phenylalkylamine type (verapamil, gallopamil) on basal and parathyroid hormone-induced bone resorption utilizing organ-cultured neonatal mouse calvaria. Only at 10(-4) M, amlodipine, verapamil and gallopamil reduced basal and parathyroid hormone-induced resorption In contrast, nifedipine, between 10(-5)-10(-4) M, exhibited a dose-dependent inhibitory effect on parathyroid hormone-related bone resorption by up to 50%. When calvariae were cultured for 48 hr in the presence of inhibitory concentrations of the calcium channel blockers and then stimulated with parathyroid hormone, only parietal bones pretreated with nifedipine remained completely responsive to the bone resorbing action of the hormone. PMID- 9225362 TI - Modelling of non-linear pharmacokinetics in sheep after short-term infusion of cardiotoxic doses of imipramine. AB - Imipramine was administered to sheep (n = 10) by intravenous infusion in high doses (450 mg-900 mg) to elicit cardiovascular shock. A cardiac assist device was then employed to manage the acute overdose situation. The concentration-time course of imipramine and its metabolite desmethylimipramine in plasma was measured by HPLC. As an indicator of imipramine's cardiotoxic effect, cardiac output was monitored. The aim of the study was to evaluate the pharmacokinetics under these conditions and to assess the efficiency of a cardiac assist device with (n = 5) and without (n = 5) an integrated haemoperfusion unit in removing drug from the circulation. The kinetics of imipramine could be described by a three compartment body model with concentration-dependent clearance resulting in non-linear kinetics. The changes in cardiac output with time could be linked to the pharmacokinetic model by a linear relationship. The cardiac assist device was found to contribute to the overall elimination of imipramine whereas the haemoperfusion unit had no clinically relevant impact. PMID- 9225363 TI - T25: a simplified carcinogenic potency index: description of the system and study of correlations between carcinogenic potency and species/site specificity and mutagenicity. AB - A simplified carcinogenic potency index, the T25, is proposed as a practical method for the inclusion of potency considerations in carcinogen classification systems. The T25 is the chronic daily dose in mg per kg bodyweight which will give 25% of the animals tumours at a specific tissue site, after correction for spontaneous incidence, within the standard life span of that species. Calculated T25 values of a set of 113 US National Cancer Institute/National Toxicology Program (NC/NTP) carcinogens showed excellent correlation (correlation coefficient 0.96, P < 0.0001) with the carcinogenic potency index TD50 of Peto et al. (1984). The mean of T25 values for 51 transspecies, multiple common site NCI/NTP carcinogens were 10-fold lower than those for 62 NCI/NTP single species, single site carcinogens. For these 113 carcinogens, the mean T25 values were approximately 3-fold lower for agents that were also mutagenic in Salmonella compared to the non-mutagenic agents. PMID- 9225364 TI - Relaxation of rat resistance arteries by acetylcholine involves a dual mechanism: activation of K+ channels and formation of nitric oxide. AB - The relaxation of rat mesenteric resistance arteries to acetylcholine was studied in vessels (normalised internal diameter 230-330 microns) mounted in an isometric myograph and contracted with noradrenaline (5 microM). Removal of the endothelium abolished acetylcholine-induced vasorelaxation, whereas pretreatment with NG nitro-L-arginine (500 microM) only inhibited the response partly. The relaxation was, however, completely inhibited by NG-nitro-L-arginine when the arteries were contracted with 80 mM K+. Acetylcholine-induced vasorelaxation was also attenuated by pretreatment with the K+ channel blocker, iberiotoxin (100 nM), and the combined pretreatment with iberiotoxin+NG-nitro-L-arginine completely blocked vasorelaxation to acetylcholine. Further, vasorelaxation to acetylcholine was attenuated by tetraethylammonium (5 mM), 4-aminopyridine (1 mM), and BaCl2 (100 microM), respectively, whereas glibenclamide (1 microM) and indomethacin (10 microM) were devoid of effect. Vasorelaxation to the nitric oxide donor sodium nitroprusside was not influenced by iberiotoxin. We conclude that in rat mesenteric resistance arteries, there is a significant nitric oxide-independent component of acetylcholine-induced vasorelaxation, which is mediated by activation of several types of K+ channels, in particular large conductance Ca(2+)-dependent K+ channels. PMID- 9225365 TI - Modulation of the megakaryoblastic Dami cell line differentiation by phosphodiesterase inhibitors and imidazo[1,2-a]pyrazine derivatives. AB - Phosphodiesterase inhibitors have been shown to modulate cell differentiation. We have previously shown that a series of imidazo[1,2-a]pyrazine derivatives displayed inhibitory effects on phosphodiesterase isoenzymes types III. IV and V isolated from Dami cells and on Dami cell growth. In the present study we have investigated the effect of these derivatives on the expression of two differentiation markers, glycoproteins Ib and IIb/IIIa of the human megakaryoblastic leukaemic Dami cell line in comparison to those elicited by 3 isobutyl-1-methylxanthine and selective phosphodiesterase inhibitors of types 1 (8-methoxymetyl-1-methyl-3-(2-methylpropyl) xanthine), III (Milrinone), IV (RO 201724) and V (Zaprinast). Imidazo[1,2-a]pyrazine derivatives, 3-isobutyl-1 methylxanthine and selective phosphodiesterase inhibitors, except 8-methoxymethyl 1-methyl-3-(2-methylpropyl) xanthine, decreased glycoprotein Ib expression. SCA40, SCA41, SCA44 and 3-isobutyl-1-methylxanthine-but not the other compounds affected the expression of glycoprotein IIb/IIIa in a positive manner. The effects of imidazo[1,2-a]pyrazine derivatives on glycoprotein expression appeared to be related to their phosphodiesterase inhibitory potency. PMID- 9225366 TI - The in vivo effects of interleukin-3 on histamine levels in non-Hodgkin's lymphoma patients. AB - Recombinant human Interleukin-3 (RhIL-3) is a haemopoietic growth factor with effect both on early and differentiated cells, such as eosinophils and basophils, and it also acts as a histamine-releasing agent. The purpose of the present study was to examine whether in vivo rhIL-3 administration after chemotherapy affected basophil histamine levels and whether a concordance between rhIL-3 induced histamine release and side effects during the treatment could be demonstrated. Thirty patients with non-Hodgkin's lymphoma entered the study. All patients received 6 courses of chemotherapy, rhIL-3 was administered subcutaneously once daily after the second and the fourth course of chemotherapy from cycle day 2-15 at the dose levels 0.5, 1.0, 5.0, 7.5 and 10 micrograms/kg with 6 patients at each dose level. In cycle 6 recombinant human Granulocyte-Macrophage Colony Stimulating Factor (rhGM-CSF) (3.0 micrograms/kg) was administered sequential/concurrent day 9-15 to rhIL-3 (day 2-15) at all dose levels except 7.5 micrograms/kg, where rhIL-3 was given day 2-8 and rhGM-CSF sequential day 9-15. Cycles 1, 3 and 5 served as control cycles with no cytokine therapy. During rhIL 3 treatment, and after CHOP chemotherapy, the basophil counts increased moderately especially during the recovery period day 15-22, and mainly at the two highest dose levels 7.5 and 10 micrograms/kg, but never exceeded the normal upper limit. Histamine levels in basophils were the same in patients before chemotherapy and healthy volunteers, and except from a trend to increased histamine level at 10 micrograms/kg on day 15, no difference was noted between rhIL-3 cycles and control cycles. Within 3-4 hr after rhIL-3 administration, a drop in histamine level in basophils was noted, which could be due to histamine releasing properties of rhIL-3 as previously demonstrated by in vitro studies. No serious side effects were noted during the cytokine treatment, and despite that most patients had mild flushing of the face, neck and upper chest, no patients experienced sensitization throughout the study. Although a significant increase in rhIL-3-induced histamine release from basophils was noted in some of the patients, no correlation to the dose of rhL-3 was found, and no correlation was noted between side effects and histamine release or histamine levels in basophils. PMID- 9225368 TI - Blood and plasma lipoprotein distribution and gender differences in the plasma pharmacokinetics of lipid-associated annamycin. AB - The objectives of this study were to determine the lipoprotein distribution of unbound annamycin and liposomal annamycin within human and rabbit blood and plasma and to evaluate the plasma pharmacokinetics of liposomal annamycin in male and female rabbits following a single intravenous bolus of the compound. Annamycin and liposomal annamycin were incubated in human and rabbit blood and plasma for 60 min. at 37 degrees C. Following incubation blood and plasma samples were assayed by HPLC for drug in each of the lipoprotein and lipoprotein deficient plasma fractions. To evaluate the plasma pharmacokinetics of liposomal annamycin in male versus female rabbits, a single intravenous bolus dose (5 mg/kg) of liposomal annamycin was administered to male and female rabbits. Sequential blood samples were obtained from the animals following the dose, analyzed for drug, and the pharmacokinetics determined using multicompartmental methods. The incorporation of annamycin into liposomes composed of dimyristoylphosphatidylcholine and dimyristoylphosphatidylglycerol resulted in no significant differences in blood versus plasma lipoprotein drug distribution. Furthermore, no differences in the plasma distribution of liposomal annamycin were observed when the drug was either incubated in vitro for 1 hr or 1 hr following intravenous administration into New Zealand male white rabbits. The plasma clearance and volume of distribution of liposomal annamycin were decreased and a increase in plasma AUC in female as compared to male rabbits following a single intravenous bolus of liposomal annamycin was observed. These findings suggest that the lipoprotein distribution of liposomal annamycin is not different when incubated in blood or plasma and that in vitro liposomal annamycin plasma distribution is similar to in vivo. Furthermore, it appears that the pharmacokinetics of liposomal annamycin are different following administration to male versus female rabbits. PMID- 9225369 TI - Cultural distance and its relationship to psychological adjustment of international exchange students. AB - This study aimed to evaluate the reliability and validity of the Japanese version of the Cultural Distance Questionnaire (CDQ), and to examine whether or not the measured cultural distance influenced the psychological adjustment of international exchange students and, if so, which aspects were most influential. Subjects were 211 Japanese high school and college students who stayed for one year with a host family in one of 23 countries around the world. They were asked to complete the Maudsley Personality Inventory before departure from Japan, the 12-item General Health Questionnaire six months after arrival in the host country and the CDQ six months after they returned from abroad. We found that the CDQ had high internal consistency reliability. Satisfactory validity was suggested for the CDQ because it offered wide coverage of various aspects of daily lives that reflect cultural differences, because it was uncontaminated by social desirability or by personality traits and because the obtained scores by country and region were largely in accordance with the expected directions. It was found that the greater the cultural distance between Japan and the foreign community, as measured by the CDQ, the greater the psychological distress of the international student placed in that community. Furthermore, the results showed that it was the food that had the greatest impact on the intercultural adjustment. PMID- 9225367 TI - On the high affinity of 8-cyclohexylcaffeine for the presynaptic inhibitory adenosine receptor present in rat motor nerve terminals. AB - Rat neuromuscular junction was used to study the characteristics of presynaptic A1 adenosine receptors. We investigated the ability of the 8-substituted caffeine, 8-cyclohexylcaffeine (CHC), as well as of 1,3,8-substituted xanthines, 1,3-dipropyl-8-p-sulfophenylxanthine (DPSPX) and 8-p-sulfophenyl-1-isoamyl-3 isobutylxanthine (SPIIBX) to antagonize the inhibitory effect of 2 chloroadenosine on the amplitude of nerve-evoked twitches of the rat phrenic hemidiaphragm, and we compared the affinity of these xanthines with that of 1,3 dipropyl-8-cyclopenthylxanthine (DPCPX). CHC, DPSPX and SPIIBX in a near parallel manner shifted to the right the log concentration-response curve for the inhibitory effect of 2-chloroadenosine on nerve-evoked twitch amplitude. Linear Schild plots with slopes near to unity were obtained for all these xanthines. The order of potency of the xanthines was DPCPX (Ki = 0.53 nM) > DPSPX (38 nM) = CHC (41 nM) > SPIIBX (404 nM). The affinities of DPSPX and SPIIBX for the A1 receptor at the rat neuromuscular junction are in agreement with the affinities described for A1 receptors at brain membranes. The now reported affinity of CHC for the presynaptic A1 receptor is 683 times higher than that obtained in binding studies in rat brain membranes, and is only 49 times higher than that obtained in functional assays (adenylate cyclase activity) in non-neuronal preparations (rat fat cells). PMID- 9225370 TI - Risk factors for postpartum depression in Japan. AB - We conducted a longitudinal study to identify risk factors for postpartum depression. At the late phase of pregnancy, 627 pregnant women agreed to take the State-Trait Anxiety Inventory Trait (STAIT) test and to remain in the study until 4 months postpartum. At 1, 3 and 4 months postpartum, they took the Edinburgh Postnatal Depression Scale (EPDS) test and the State-Trait Anxiety Inventory State (STAIS) test. At 3 months postpartum, they were asked about socio psychological and obstetric factors. High scores in the EPDS and STAIS tests were correlated with primiparity, premature delivery, difficult labor, experience of life events and worries about baby care. Furthermore, high scores in the STAIT test in late pregnancy were strongly correlated with high scores in the EPDS and STAIS tests in the postpartum period. PMID- 9225372 TI - Periodic leg movements in sleep in essential hypertension. AB - The presence of periodic leg movements in sleep (PLMS) was assessed in 91 subjects diagnosed with essential hypertension. More than 18 per cent of the sample had PLMS, which is considerably higher than in normal controls. Also, the prevalence was significantly correlated with the severity of hypertension, as well as with age. Periodic leg movements in sleep were more frequent in the first few hours of the sleep period and during sleep stages 1 and 2. The arousing effect of PLMS was minimal, with only 17 per cent of all events related to an EEG arousal. Our results suggest that PLMS are common in people with essential hypertension, although they do not seem to be associated with any particular sleep disorder. PMID- 9225371 TI - Recurrent paroxysmal episodes characterized by perceptual alteration in three schizophrenic patients on neuroleptic medication. AB - A suddenly occurring episode characterized by perceptual alteration (SEPA), mainly of visual and/or auditory modalities, which repeatedly occurred in three schizophrenic patients on long-term neuroleptic medication, is described. Perceptual alteration showed some distinct features that were different from acute symptoms of schizophrenia, and was accompanied by mood changes such as severe anxiety and agitation and, in one of the patients, also by extrapyramidal symptoms. Perceptual alteration, as well as mood changes and extrapyramidal symptoms, responded well to an anticholinergic drug, biperiden. Recent studies have shown that SEPA occurred not only in schizophrenic patients but also in patients on long-term neuroleptic medication for treating other mental disorders. These findings suggest that SEPA is associated with dopaminergic hypoactivity in the brain, which is induced by long-term neuroleptic medication. PMID- 9225373 TI - Free-running circadian rhythm of melatonin in a sighted man despite a 24-hour sleep pattern: a non-24-hour circadian syndrome. AB - Sleep and plasma melatonin rhythms were measured longitudinally in a sighted young man (21 years old) under a day-right environment. At each measurement, the responsiveness of the melatonin rhythm to a single light pulse was examined in addition to the 24-hour profile. In experiment 1, the timing of sleep was decided by the subject himself. Although most sleep episodes were observed between 21:02 h and 10:55 h, the plasma melatonin rhythm free-ran for a period of 24.18 h. In experiment 2, the sleep-wake schedule of the subject was strictly fixed. The subject was instructed to go do bed at 24:00 h and wake up, at the latest, before 8:00 h for 40 days. The melatonin rhythm, however, continued to free-run for a period of 24.12 h. Nocturnal melatonin level could not be suppressed by a 3-hour light pulse of 500 lx, but was suppressed by a pulse of 1000 lx. It is concluded that internal desynchronization occurred in this particular sighted subject where the sleep-wake rhythm was entrained by the 24-hour day-night environment, whereas the plasma melatonin rhythm free ran, and that a forced sleep schedule did not act as a strong zeitgeber. PMID- 9225374 TI - A sighted man with non-24-hour sleep-wake syndrome shows damped plasma melatonin rhythm. AB - Twenty-four-hour profiles of plasma melatonin, cortisol and rectal temperature were measured longitudinally in a sighted man who has been suffering from sleep disorders for more than 10 years. The sleep-wake rhythm of this subject free-ran, despite his routine life, and occasionally showed a sign of internal desynchronization, where sleep was lengthened up to 30 h. These states were classified into the non-24-hour sleep-wake syndrome. Plasma melatonin concentrations in the subjective night remained at a low level and showed a damped circadian rhythm. At the same time, robust circadian rhythms were detected in plasma cortisol and rectal temperature, indicating that the circadian pacemaker was intact. The causal relationship between the damping of nocturnal melatonin rise and a failure of entrainment of the sleep-wake cycle is discussed. PMID- 9225375 TI - Daily melatonin intake resets circadian rhythms of a sighted man with non-24-hour sleep-wake syndrome who lacks the nocturnal melatonin rise. AB - Effects of daily melatonin intake on the circadian rhythms of sleep and wakefulness, rectal temperature and plasma cortisol were examined in a sighted man who had suffered from the non-24-hour sleep-wake syndrome. The subject lacked the nocturnal melatonin rise in plasma, but showed robust circadian rhythms in rectal temperature and plasma cortisol. The sleep-wake rhythm free-ran with a period longer than 24 hours. Daily melatonin intake at 21:00 h concentrated sleep episodes in the nocturnal period (24:00-8:00 h), and increased the length of the episodes. A single oral dose (3 mg) of melatonin increased plasma melatonin levels to about 1300 pg/mL within one hour and remained at pharmacological levels for approximately 6 hours. The trough of rectal temperature and the circadian rise of plasma cortisol were fixed to the early morning. A higher dose of melatonin (6 mg) did not improve the general feature. After the cessation of melatonin intake, the sleep-wake rhythm began to free-run together with the circadian rhythms in rectal temperature and plasma cortisol. It is concluded that daily intake of melatonin at early night time resets the circadian rhythms in a sighted man who lacked the nocturnal melatonin rise and showed free-running circadian rhythms in routine life. PMID- 9225376 TI - Fluctuations of rectal and tympanic temperatures with changes of ambient temperature during night sleep. AB - Many studies have demonstrated a decline in core temperature during slow wave sleep (SWS) in animals and humans. However, there are few studies that have investigated core temperature fluctuation during rapid eye movement sleep (REM) at different ambient temperatures. This study examined the effects on core temperature of continuous hot or cold exposure during sleep. Ten male subjects were exposed to hot and cold stress from 00 h to 1:00 h, when SWS is most predominant, and from 5:00 h to 7:00 h, when REM is predominant. Rectal temperature (Tr) and tympanic temperature (Tt) were monitored for 3 days, and polysomnographies (PSG) were recorded from 23:00 h to 7:00 h. The experiments lasted 3 weeks for each subject, over 2 consecutive nights each week (including an adaptation night and an experimental night). On the experimental night, subjects were exposed to hot (29 degrees C) or cold (21 degrees C) ambience. The core temperature fluctuation under the hot or cold ambience were compared with under the thermoneutral ambience. Under hot ambience, Tr declined significantly in the first 2 hours of sleep, but Tt did not change. In the last 2 hours, both Tr and Tt were significantly elevated. Under cold ambience, both Tr and Tt declined significantly in the first 2 hours. However, in the last 2 hours, neither Tr not Tt showed any change. The result that Tr and Tt rose in hot ambience during the last 2 hours when REM is predominant suggests that body temperature during REM is influenced by ambient temperature. PMID- 9225378 TI - Influence of reference electrodes, stimulation characteristics and task paradigms on auditory P50. AB - To clarify the nature of auditory P50, middle latency auditory evoked potentials were recorded by using different conditions of reference electrodes (linked earlobes, LE; balanced non-cephalic, BN), stimulation characteristics (tone burst, human voice) and tasks (counting, simple reaction) in 10 right-handed males (aged 21-36 years). EEG was recorded from Fz, Cz, Pz, C3, C4, T3 and T4 according to the 10-20 system. Two groups of electrode sites were made for the statistical analysis: a midline group, Fz, Cz and Pz, and a lateral group, T3, C3, Cz, C4 and T4. The results were that the P50 amplitudes with BN electrodes were significantly higher than those with LE in both groups (midline, P < 0.01; lateral, P < 0.01); the P50 amplitudes by voice stimulation were significantly higher than those by tone stimulation in the lateral group (P < 0.05), and the P50 latencies under a simple reaction paradigm were significantly shorter than those under a counting task in both groups (midline, P < 0.05; lateral, P < 0.05). These results suggest that various factors including motor response affect the P50 amplitudes and latencies. PMID- 9225377 TI - Identification of monocyte chemoattractant protein-1 in senile plaques and reactive microglia of Alzheimer's disease. AB - It has been shown that human monocytes express monocyte chemoattractant protein-1 (MCP-1), an inflammatory factor, in response to non-fibrillar beta-amyloid protein. Reactive microglia and inflammatory factors were reported to be present in beta-amyloid deposits (senile plaques) in Alzheimer's disease, suggesting the presence of MCP-1 in senile plaques. To address this issue, we examined MCP-1 immunoreactivity in senile plaques using a mouse monoclonal anti-MCP-1 antibody. Monocyte chemoattractant protein-1 was found immunohistochemically in mature senile plaques and reactive microglia but not in immature senile plaques of brain tissues from five patients with Alzheimer's disease. These findings suggest that MCP-1-related inflammatory events induced by reactive microglia contribute to the maturation of senile plaques. PMID- 9225379 TI - Polarization anatomy of a kainic acid seizure. AB - Slow voltage-sensitive dyes work by accumulating in brain tissue and report the average membrane potential of neurons and glia. The voltage-sensitive dye diO-C2 5 was used to monitor the polarization state of 27 brain structures in the rat during a systemically induced, behaviorally mild, kainic acid seizure using a 20 s recording period. The effects of the anesthetic agent used in the experiment were minimized by delaying the dye injection and seizure mapping for one day. Eleven areas were depolarized during the seizure, but 16 other areas did not change their polarization state compared to controls. The effects of pentobarbital appear to have no measurable effect on seizure propagation once the animal has behaviorally recovered from the anesthesia. The technique allows for mapping areas of seizure involvement with a unique combination of spatial and temporal resolution. PMID- 9225380 TI - Nuclear polyadenylate polymerase activity in the brain of seizure-prone EL mice. AB - Nuclear polyadenylate polymerase from I activity in the brains of seizure-prone EL mice was significantly higher than in seizure-non-susceptible progenitor ddY mice. This finding may be essential in acquiring susceptibility to seizures, since there was no significant difference between EL(S) mice and those that did not receive stimulation, EL(NS) mice. Lower form II enzymatic activity was observed in both groups of EL mice but not in ddY mice. Moreover, significantly lower activities of form II 7 days after seizures were found in EL(S) mice compared with EL(NS) mice, suggesting that this is a consequence of repeated seizures. The activity of form I enzyme decreased immediately and at 30 and 60 min after seizures, then returned to control levels at 100 min. Form II enzymatic activity was significantly decreased only at 30 min after seizures, implying that seizures exerted a later effect on form II enzyme. These changes may cause a decrease in the rate of polyadenylation in the brain; thus, alteration of post transcriptional events, including messenger RNA processing and transport, may occur during epileptic seizures. PMID- 9225381 TI - Temporal and regional profiles of cytoskeletal protein accumulation in the rat brain following traumatic brain injury. AB - To characterize the cytoskeletal aberration due to traumatic injury, temporal and regional profiles of changes in immunoreactivity of microtubule-associated protein 2 (MAP2), neurofilament heavy subunit protein (NFH) and heat shock protein 72 (HSP72) were investigated after different magnitudes of traumatic brain injury by fluid percussion. The experimental rat brain was perfusion-fixed at 1, 6 and 24 hours after traumatic brain injury. Conventional histological staining has demonstrated that the mildest traumatic brain injury (1.0 atm) induced no neuronal loss at the impact site and that neuron loss was apparent when traumatic brain injury was increased to 4.3 atm. The mildest traumatic brain injury, however, caused a significant increase in HSP72 immunoreactivity in the superficial cortical layers at the impact site as early as 1 hour after the injury. In the case of severe traumatic brain injury (4.3 atm), neuron loss was apparent in the area at the impact site, but the increase in HSP72 immunoreactivity was moderate, and it was observed only after 6 hours in the deep cortical layers under the necrotic area. The increased immunostaining of MAP2 was demonstrated in damaged axons and neuronal perikarya in the wider area surrounding the impact site at 6 and 24 hours after the injury. Six and 24 hours after the injury, perikaryal accumulation of neurofilament was observed, and the accumulated neurofilament was mostly phosphorylated. These results indicate that the severe traumatic brain injury of 4.3 atm triggers the abnormal accumulation of cytoskeletal proteins in neuronal perikarya, most probably due to an impairment of axonal transport. It is implied that the increased expression of HSP72 may be involved in the protective process of neurons after traumatic brain injury. PMID- 9225382 TI - Strain difference in behavioral response to a new environment in rats. AB - Locomotor activity in rats throughout a 24-hour period in a new environment was examined for strain differences and for the capacity for adaptation to that environment. Fischer 344 rats (F344), spontaneously hypertensive rats (SHR) and Wistar normotensive Kyoto rats (WKY) were used. The horizontal locomotor activity of individual rats was measured by photocell-utilizing activity-recording devices. The locomotor activity counts on the second day, after 1 day of adaptation, were compared with those after 5 days of adaptation (on the sixth day). In WKY, there was no difference in activity at any period of the day between the second and sixth days. In SHR, the locomotor activity on the second day between 6:00 h and 9:00 h (in the light phase) and between 24:00 h and 3:00 h (in the dark phase) was higher than on the sixth day. In F344, the locomotor activity on the second day between 18:00 h and 2:00 h was higher than on the sixth day. The capacity for adaptation in SHR and F344 was thus poorer than in WKY. The poor adaptation in SHR and F344 was similar to that in depressive patients induced by moving house. These findings suggested that SHR or F344 were suitable for depression research. PMID- 9225383 TI - Negative correlation of reaction times and the auditory P50 in a Go, No-Go paradigm. AB - We investigated the relationship between reaction times in a Go, No-Go paradigm and various components of the auditory event-related potentials. The P50 latencies in the Go trials were significantly earlier than those in the No-Go trials. The N200 and P300 were positively associated with the reaction times. However, the P50 latencies showed negative correlation with the reaction times, and the negative correlation at F3 was the largest. Referring to source studies on P50, it is suggested that the activation of either the ascending activating system or the left premotor area through that system might affect the speed of reaction. PMID- 9225384 TI - Slipped capital femoral epiphysis. AB - SCFE is the most common hip abnormality in adolescence. The subsequent development and severity of degenerative changes is related to the degree of slippage and to delay in diagnosis. Awareness of this diagnostic possibility in the population at greatest risk and knowledge of subtle and early radiographic findings will allow prompt diagnosis and treatment of SCFE. Follow-up radiographic examination should be used to evaluate for physeal fusion and any complications, including progression of the slip, hardware complications, chondrolysis, avascular necrosis, and secondary osteoarthritis. PMID- 9225386 TI - Subtle or atypical injuries of the thoracic aorta and brachiocephalic vessels in blunt thoracic trauma. AB - Aortic or brachiocephalic vessel injuries secondary to blunt thoracic trauma are relatively common and can occur throughout the length of the thoracic aorta or in various locations in the brachiocephalic vessels. Aortography remains the standard of reference for the diagnosis of these injuries despite recent technologic advances in other imaging modalities. The classic aortographic finding in aortic or brachiocephalic vessel injury consists of a large false aneurysm, typically protruding from the medial aspect of the aortic isthmus. However, intrathoracic aortic or brachiocephalic vessel injury can and does occur at any intrathoracic location and may exhibit a wide variety of radiographic appearances, thereby presenting a diagnostic challenge even for experienced trauma angiographers. Large false aneurysms may appear oval or rounded, tubular, or asymmetrically globular and may manifest in unusual locations such as the ascending aorta. Although smaller, irregularly shaped false aneurysms at atypical locations may be obscure or mimic ductus diverticula, their irregular, sharp margins allow them to be distinguished as injuries. The subtlety of aortic or brachiocephalic vessel injuries necessitates a high degree of suspicion along with meticulous imaging technique in all cases and the use of additional projections in equivocal cases for definitive diagnosis. PMID- 9225385 TI - "Lumps" and "bumps" that mimic acute aortic and brachiocephalic vessel injury. AB - Laceration of the thoracic aorta or brachiocephalic vessels due to blunt trauma is relatively common. In such cases, prompt and accurate diagnosis followed by timely surgery is essential. These injuries typically occur at the aortic isthmus and can usually be readily identified at aortography, which remains the standard of reference for diagnosis. However, numerous anatomic variants that manifest as "lumps" or "bumps" on aortograms can mimic true vascular injury, thereby leading to false-positive or false-negative diagnosis. These variants include aortic spindle, classic or atypical ductal diverticula, and infundibula of the brachiocephalic arteries and adjacent branches or of the right third intercostal artery. Ductus diverticula typically occur at the isthmus and have smooth, uninterrupted margins with gently sloping shoulders. Infundibula are also smoothly marginated but can occur in a variety of locations and generally taper into one or more vessels at their apex. Knowledge of the imaging appearances of these anatomic variants is necessary for correct interpretation of aortograms of the aorta and brachiocephalic vessels in blunt trauma patients. PMID- 9225387 TI - CT of renal inflammatory disease. AB - Although computed tomography (CT) is not routinely indicated in uncomplicated renal infection, it is of value in establishing the diagnosis in equivocal cases, in evaluating high-risk patients, and in determining the extent of disease. Unenhanced CT is useful in demonstrating gas, calculi, parenchymal calcifications, hemorrhage, and inflammatory masses. However, a contrast material enhanced study is essential for complete evaluation of patients with renal inflammatory disease to demonstrate alterations in renal excretion of contrast material that occur as a result of the inflammatory process. In severe acute pyelonephritis, enhanced CT scans obtained during the cortical nephrographic phase typically demonstrate solitary or multifocal areas of hypoattenuation with loss of the corticomedullary interface. Delayed CT scans obtained during the excretory phase are frequently more helpful than early CT scans in defining the extent of the disease process, identifying complications such as renal abscess, and confirming the presence of urinary obstruction. PMID- 9225389 TI - Evaluation of the portal venous system: complementary roles of invasive and noninvasive imaging strategies. AB - Evaluation of the portal venous system is required in several clinical circumstances, including before and after liver transplantation, before creation of a transjugular intrahepatic portosystemic shunt, in the clinical setting of bowel ischemia, or to evaluate varices. Several noninvasive modalities (magnetic resonance [MR] imaging and MR angiography, computed tomography [CT], and ultrasound [US]) are available for evaluation of the portal venous system in addition to the invasive angiographic methods. In most clinical circumstances, either CT or MR imaging and MR angiography in combination with US of the liver vasculature will allow complete evaluation of the portal venous system. Invasive evaluation of the portal venous system is necessary when results of the noninvasive tests disagree or are inconclusive. Angiography may also be indicated whenever noninvasive tests indicate occlusion of the portal venous system, as this is often a crucial clinical question and false-positive results can occur with the noninvasive tests. PMID- 9225388 TI - Complications of peritoneal dialysis: evaluation with CT peritoneography. AB - Computed tomographic (CT) peritoneography involves CT of the abdomen and pelvis after administration of a mixture of contrast material and dialysate. CT peritoneography can demonstrate a variety of complications of continuous ambulatory peritoneal dialysis. In patients with symptoms of peritonitis, CT peritoneography is better than conventional CT in demonstrating loculated fluid collections and indicates adhesions by means of uneven distribution of the contrast material-dialysate mixture. In patients with edema or abdominal bulging, CT peritoneography reliably shows the site of dialysate leakage and allows differentiation of a leak from a hernia. In patients with problems of fluid return, catheter malposition and its effect on dialysate distribution can be determined with CT peritoneography. In patients with poor ultrafiltration, demonstration of restricted space in the pelvis or poor distribution of fluid with CT peritoneography suggests adhesions. CT peritoneography also provides anatomic information for referring physicians that may determine whether treatment is medical or surgical. PMID- 9225390 TI - CT of acquired abnormalities of the portal venous system. AB - Computed tomography (CT), including biphasic contrast material-enhanced helical dynamic scanning and three-dimensional CT angiography, is useful in evaluating acquired abnormalities of the portal venous system. At contrast-enhanced CT, portal venous thrombus usually manifests as low-attenuation intraluminal lesions combined with enlargement of the affected portal vein. Cavernous transformation, a masslike network of intertwined veins that provides an alternative pathway for a stenosed or occluded portal vein, is depicted as multiple, periportal vascular structures. At helical dynamic CT, arterioportal shunts manifest as early enhancement of the affected portal vein, transient hyperperfusion abnormalities with lobar or segmental distribution, or transient wedge-shaped enhancement peripheral to the tumor. In patients with portosplenic venous invasion by malignant neoplasms, peripancreatic or perigastric veins may dilate if they function as hepatopetal collateral veins. In patients with portal hypertension, a variety of hepatofugal collateral pathways can develop, including esophageal, paraesophageal, coronary gastric, inferior phrenic, paraumbilical, abdominal wall, splenorenal, gastrorenal, retrocaval, and mesocaval collateral pathways. An understanding of the varied CT appearances of acquired abnormalities of the portal venous system will allow more definitive diagnosis and help avoid false diagnosis of disease. PMID- 9225391 TI - From the archives of the AFIP. Genitourinary rhabdomyosarcoma in children: radiologic-pathologic correlation. AB - Rhabdomyosarcoma is the most common tumor of the lower genitourinary tract in children in the first 2 decades of life. Most cases of genitourinary rhabdomyosarcoma are of the embryonal histologic subtype and include tumors of the bladder, prostate, testes and paratesticular sites, penis, perineum, vagina, and uterus. The natural history, pattern of metastatic spread, treatment, and prognosis of childhood rhabdomyosarcoma vary with the anatomic site of the lesion. In children with rhabdomyosarcoma of the bladder or prostate, presenting signs and symptoms include urinary or fecal retention, dysuria, urinary tract infection, and hematuria. Paratesticular rhabdomyosarcoma produces painless scrotal swelling, which may be ignored until the tumor has reached a large size. Vaginal tumors may manifest as a prolapsing mass in the introitus. Radiologic studies of children with genitourinary rhabdomyosarcoma reflect the nonspecific gross features of the tumor, which may be ill defined with infiltrative margins or well circumscribed by a pseudocapsule of compressed tissue. The botryoid variant of embryonal rhabdomyosarcoma results when submucosal tumor produces a polypoid mass resembling a cluster of grapes within a hollow structure. Botryoid morphology is characteristic, but not specific, for rhabdomyosarcoma within the vagina or urinary bladder, since yolk sac tumor and "tumoral" cystitis may have a similar appearance. Invasion of adjacent structures by the primary tumor may make the precise anatomic origin of genitourinary rhabdomyosarcoma difficult to determine on cross-sectional images. Recent refinements in multidisciplinary therapeutic regimens combining chemotherapy, radiation therapy, and surgery have dramatically improved outcome for children with genitourinary rhabdomyosarcoma. Diagnostic imaging plays an important role in monitoring response to therapy. PMID- 9225392 TI - From the RSNA refresher courses. Challenges of pediatric spiral CT. AB - Spiral technology has expanded the usefulness of computed tomography (CT) in the evaluation of pediatric diseases. Even though spiral CT requires shorter total scanning times, image degradation by patient motion is still a problem in very young patients, and oral and intravenous sedatives are administered to minimize this problem. Optimal parenchymal enhancement depends on the amount and injection rate of contrast material and the timing of the onset of scanning. All these are more variable in studies of children than in adults, with the latter being the most complicated. Spiral CT of the pediatric chest is most useful for evaluating anastomotic dehiscence and stenosis in lung transplant recipients, pulmonary nodules in children with malignant disease likely to disseminate to the lung, and great vessel anomalies; for staging pulmonary tumors; and for dynamic imaging to identify various diseases that cause pulmonary dysfunction. Important applications of spiral CT of the pediatric abdomen and pelvis include evaluating the liver for acquired vascular abnormalities and vascular tumors, staging and preoperative assessment of renal tumors, and evaluating inflammatory pelvic lesions. PMID- 9225393 TI - The AAPM/RSNA physics tutorial for residents. X-ray production. AB - X-rays are produced when highly energetic electrons interact with matter and convert their kinetic energy into electromagnetic radiation. The two unique mechanisms by which x-rays are produced are called the bremsstrahlung and characteristic processes. Bremsstrahlung x-rays produce a continuous x-ray spectrum, whereas characteristic x-rays are produced at specific narrow bands of energies. Many technical parameters of the x-ray production equipment affect the magnitude and shape of the x-ray spectrum. The quantity of x-rays produced varies proportionally to the tube potential squared, tube current, exposure time, and atomic number of the anode material and is inversely proportional to the distance squared. x-ray quantity is also affected by the voltage waveform (generator type) and tube filtration. The shape of the x-ray spectrum is affected by the atomic number of the anode material, tube potential, filtration, and voltage waveform. PMID- 9225394 TI - Primer on computers and information technology. Part one: an introduction to the computer. PMID- 9225395 TI - Primer on computers and information technology. Part two: an introduction to computer networking. AB - Computers networks are a way of connecting computers together such that they can exchange information. For this exchange to be successful, system behavior must be planned and specified very clearly at a number of different levels. Although there are many choices to be made at each level, often there are simple decisions that can be made to rapidly reduce the number of options. Planning is most important at the highest (application) and lowest (wiring) levels, whereas the middle levels must be specified to ensure compatibility. Because of the widespread use of the Internet, solutions based on Internet technologies are often cost-effective and should be considered when designing a network. As in all technical fields, consultation with experts (ie, computer networking specialists) may be worthwhile. PMID- 9225396 TI - Residents' teaching files. Papillary renal cell carcinoma: diagnostic dilemma of a cystic renal mass. PMID- 9225397 TI - Pediatric case of the day. Mesenteric plexiform neurofibroma in neurofibromatosis type 1 (von Recklinghausen disease). PMID- 9225398 TI - US case of the day. Gastric polyp in conjunction with poorly differentiated gastric adenocarcinoma. PMID- 9225399 TI - General case of the day. Longitudinal stress fracture of the left femoral diaphysis and stress reaction in the right femur. PMID- 9225400 TI - Breast imaging case of the day. Recurrent infiltrating ductal carcinoma and radiation-induced dystrophic calcifications. PMID- 9225401 TI - Idiopathic interstitial pneumonias: high-resolution CT and histologic findings. PMID- 9225402 TI - Imaging of emphysema and lung volume reduction surgery. PMID- 9225403 TI - CT of acute pulmonary emboli: where does it fit? PMID- 9225404 TI - Complex disease of the pleural space: the 10 questions most frequently asked of the radiologist--new approaches to their answers with CT and MR imaging. PMID- 9225405 TI - Tuberculosis and atypical mycobacterial infections in the 1990s. PMID- 9225406 TI - Is non-metropolitan residence a risk factor for poor birth outcome in the U.S.? AB - The association between non-metropolitan residence and the risk of poor birth outcome in the United States was examined using the records of 11.06 million singleton births in the United States between 1985 and 1987. Rates of neonatal and post-neonatal death, low birth weight and late prenatal care among non metropolitan residents were compared to the rates among metropolitan residents. The association between residence in a non-metropolitan area and the risk of poor birth outcome was assessed in national and state level regression analyses. Residence in a non-metropolitan county was not found to be associated with increased risk of low birth weight or neonatal mortality at the national level or in most states, after controlling for several demographic and biological risk factors. Non-metropolitan residence was associated with greater risk of post neonatal mortality at the national level. Non-metropolitan residence was strongly associated with late initiation of prenatal care at both the national level and in a majority of the states. Residence in non-metropolitan areas does not appear to be associated with higher risk of adverse birth outcome. Regionalization of perinatal care and other changes in the rural health care system may have mitigated the risk associated with residing in areas relatively isolated from tertiary care. High levels of late prenatal care among non-metropolitan residents suggest a continuing problem of access to routine care for rural women and their infants that may be associated with higher levels of post-neonatal mortality and childhood morbidity. PMID- 9225407 TI - Medication, chronic illness and identity: the perspective of people with asthma. AB - The issue of compliance with prescribed medication has traditionally been dominated by the perspective of the health professional although increasingly sociologists, using qualitative methods, have begun to present the patients' point of view. However, little has been published on asthma, despite the numbers of people suffering from this chronic condition and the amount of medication regularly prescribed. This paper focuses on the perspective of a sample of S. Wales (U.K.) asthma patients who have all been prescribed prophylactic medication in the last 12 months and explores their attitudes to medication in the context of their everyday lives, using inductive qualitative research methods. Two main groups were identified: the deniers and the accepters. They differed fundamentally in their readiness to accept the identity of asthma sufferer which, in turn, was associated with very different beliefs about the nature of their problem and the meaning of the medication prescribed for it. There was also marked differences in their strategies of self-presentation and disclosure and their pattern of medication use, particularly for prophylactic medication. A third group, the pragmatists, were also identified as a possible sub-group of the accepter category who are less open within self-presentation and less consistent in their beliefs about asthma but do not reject the label entirely. Identity work, i.e. the way the respondents interpreted the social identity of asthma sufferers and managed to reconcile it with other social identities, is proposed as the most useful way of understanding the observed variation in the way people diagnosed as asthmatic conceptualise and use their medication. PMID- 9225408 TI - Health care and consumer choice: medical and alternative therapies. AB - This paper reports on research conducted in a large Canadian city during 1994 1995. The study examines the motivations of patients who choose to seek care from one of five different types of practitioners: family physicians, chiropractors, acupuncturists/traditional Chinese doctors, naturopaths and Reiki practitioners. We use the Andersen socio-behavioural model to help explain why people choose orthodox medicine or a type of alternative care. The data are derived from face to face interviews with 300 patients: 60 from each of the five modes of treatment. The findings demonstrate that this model can explain the use of alternative as well as orthodox medical services. Patients choose specific kinds of practitioners for particular problems, and some use a mixture of practitioners to treat a specific complaint. The choice of type of practitioner(s) is multidimensional and cannot solely be explained either by disenchantment with medicine or by an "alternative ideology". PMID- 9225409 TI - Socioeconomic inequity in health care: a study of services utilization in Curacao. AB - The aim of this study is to examine whether there is socioeconomic equity in health care utilization in Curacao, Netherlands Antilles. We explore how educational level is related to utilization of GPs, specialists, hospitals; dentists and physiotherapists, taking into account the effects of sex, age and inequalities in health. The study also examines whether these relationships vary according to the unit of analysis: probability (or incidence) of services use versus overall volume of contacts. The data were derived from the Curacao Health Study, a health interview survey among a random sample (N = 2248) of the non institutionalized population aged 18 and over. The results indicate that there is socioeconomic inequity in the probability of health care utilization in Curacao. People with a higher educational level are more likely to consult a specialist, dentist or physiotherapist, and are also more likely to be hospitalized. This is not only the case when the mediating effects of socioeconomic inequalities in health (need) are taken into account, but also before adjustment for health inequalities. In other words: there appears to be both vertical inequity (i.e. greater needs for services are not met by greater use) and horizontal inequity (i.e. similar needs for care are not met by similar levels of services use). The observed inequalities in use of specialists and hospitals contrast with findings from international research. The volume of health services use (i.e. the numbers of consultations) appears to be hardly connected with a person's position in the SES hierarchy; only dental services are used more extensively by higher educated individuals. PMID- 9225410 TI - Quality of life: a dynamic construct. AB - The principle of Einstein's theory of special relativity is that an observer of an apparently moving body cannot be sure if the body really has moved, if he/she has moved or if both events have occurred. Although Einstein was discussing physical events, a similar hypothesis may apply to quality of life. When using quality of life instruments, one presumes that the point of reference (the observer in Einstein's terms) does not move, i.e. that an individual's attitude towards a particular construct will remain stable. Otherwise, changes in response to particular variables cannot be interpreted. However, attitudes are not constant: they vary with time and experience and are modified by such psychological phenomena as adaptation, coping, expectancy, optimism, self-control and self-concept. For example, eating problems may be extremely important at one point in a person's life. However, when oral discomfort has been diagnosed as cancer and treated with surgery or radiation, the same individual may "objectively" demonstrate more problems when eating, but report them as less because they have now become relatively unimportant. Furthermore, paradoxical reports that some groups of ill individuals rate their quality of life higher than do "healthy" persons raise similar questions concerning between-group point of reference differences. Investigators in the fields of organisational management, education and psychology have developed techniques such as "then ratings", saliency indicators and individualised questionnaires in attempts to quantify within-subject variability and between-group differences pertaining to point of reference. We suggest that similar methods may help us to measure change in the impact of the different items of quality of life instruments. In this paper, we will describe the theories of change associated with quality of life measurement. In addition, we will present evidence suggesting that the point of reference does change, the reasons for this and possible solutions to the problem. PMID- 9225411 TI - Sex differences in physical symptoms: the contribution of symptom perception theory. AB - Health surveys, studies on physical symptom reporting, and medical registration of physical complaints find consistent sex differences in symptom reporting, with women having the higher rates. By and large, this female excess of physical symptoms is independent from the symptom measure, response format and time frame used, and the population under study. As most studies concern healthy individuals, the sex difference can not simply be attributed to a greater physical morbidity in women. In this paper we propose a number of explanations for this phenomenon, based on a biopsychosocial perspective on symptom perception. We discuss a symptom perception model that brings together factors and processes from the extant literature which are thought to affect symptom reporting, such as somatic information, selection of information through attention and distraction, attribution of somatic sensations, and the personality factors somatisation and negative affectivity. Finally, we discuss the explanations for sex differences in physical symptoms that arise from the model. PMID- 9225412 TI - Population growth, poverty and health. AB - One of the most popular explanations for the many problems that face Africa is population growth. Africa's population has doubled since 1960. Africa has the highest fertility rate in the world and the rate of population growth is higher than in any other region. At the same time, Africa faces a social and economic situation that is viewed by many as alarming. Among the problems that devastate Africa is that of persistent poor health. Africa has lower life expectancy, higher mortality rates and is affected by more disease and illness conditions than any other region. Focusing on sub-Saharan Africa, this paper examines the relationship between population growth, poverty and poor health. While most analyses have focused on population growth as an original cause of poverty and underdevelopment, this paper argues that while both population growth and poor health play a significant role in exacerbating the problem of poverty, they are themselves primary consequences of poverty rather than its cause. PMID- 9225413 TI - Appropriateness in health care: application to prescribing. AB - To help account for and address observed variations in medical practice, evaluations of "appropriateness" have sought to supplement incomplete evidence with professional opinion. This article contributes to an understanding and refinement of the construct of appropriateness by discussing how it has been defined and applied in studies of health care in general and prescribing in particular. We suggest that appropriateness is the outcome of a process of decision-making that maximises net individual health gains within society's available resources. This definition distinguishes between (in)appropriate prescribing, as an outcome, and (ir)rational prescribing as a process. To assess appropriateness, we advocate combining explicit criteria with independent review in cases of uncertainty and disagreement. Refinements based on reviews using implicit criteria should draw on shared professional knowledge and post hoc state the process followed as explicitly as possible. The Medication Appropriateness Index is shown to provide a solid foundation for identifying dimensions of prescribing appropriateness. PMID- 9225414 TI - Migrancy, masculine identities and AIDS: the psychosocial context of HIV transmission on the South African gold mines. AB - Levels of HIV infection are particularly high amongst migrant workers in sub Saharan Africa. This paper presents a case study of one such vulnerable group of migrants-underground workers on the South African gold mines-and highlights the psychosocial context of HIV transmission in the mining setting. On the assumption that social identities serve as an important influence on peoples' sexual behaviour, the study examines the way in which miners construct their social identities within the parameters of their particular living and working conditions. It also identifies some of the key narratives used by miners to make sense of their experience in the realms of health, ill-health, HIV and sexuality. Masculinity emerged as a leading narrative in informants' accounts of their working life, health and sexuality, and the paper examines the way in which the construction of masculine identities renders miners particularly vulnerable to HIV. The implications of these findings for HIV educational interventions are discussed. PMID- 9225415 TI - Cleaning the womb: constructions of cervical screening and womb cancer among rural black women in South Africa. AB - In South Africa problems with current cervical screening uptake, including low coverage and loss of screened women to follow-up, have been identified. This paper presents the findings of an anthropological study of rural Black women's perceptions and understandings of cervical symptomatology, screening and cancer conducted among three different language groups in South Africa. The data collected indicate that women were screened when presenting with symptoms of reproductive tract infection, with the result that for many the smear came to be associated with the diagnosis and treatment of sexually transmitted diseases (STDs). In some cases the smear was said itself to "clean" the womb. The results were often interpreted by women as signifying womb "dirtiness" and confirming the presence of symptomatic reproductive disease for which they had initially presented to the biomedical facility. Several barriers to screening were identified including fear of vaginal exposure, expectation of pain, being asymptomatic, and gender of the practitioner. In addition women perceived womb cancer to be invariably terminal, knowledge which was constructed from personal and community experience of the illness. The illness was closely associated with (usually female) "promiscuity". The authors discuss the implications of the data for healthworkers and health promotion specialists, in particular the association of the smear with STDs, the way in which women are recruited for screening, the perceived terminality of womb cancer, and the processes by which local knowledge about illness is constructed. The findings demonstrate the importance of medical anthropology in contributing towards the provision of effective and locally appropriate healthcare. PMID- 9225417 TI - Female life expectancy, gender stratification, health status, and level of economic development: a cross-national study of less developed countries. AB - A number of studies have attempted to account for cross-national differences in life expectancy, but relatively few have focused on female life expectancy, and even fewer on the relevance of predictors linked to gender stratification theory. The present study seeks to assess the utility of gender stratification theory in accounting for cross-national differences in female life expectancy in less developed countries. An incremental model building strategy is used to develop a final model that combines predictors linked to both industrialism theory and gender stratification theory. The analysis is based on multiple regression and cross-sectional samples that vary in size from 40 to 97 countries. Evidence is presented that several aspects of women's status have a positive effect on female life expectancy. Indicators of women's educational status, women's economic status, and women's reproductive autonomy all prove to be important predictors of female life expectancy. Analysis of interaction effects suggests that the strength of the effects of some aspects of women's economic status and the effect of some aspects of health status on female life expectancy vary with the level of economic development. A comprehensive assessment of the relative strength of alternative measures of women's education is carried out, and evidence is presented that it does make a difference how the level of women's education is measured. PMID- 9225416 TI - Head injury rehabilitation in the U.K.: an economic perspective. AB - The human and societal costs as a result of traumatic brain injury (TBI) are extensive with approximately 200-300/100,000 of the population requiring hospitalisation each year in the U.K. Advances in neurosurgical management have meant that more people sustaining head injuries are surviving. The need for rehabilitation programmes for these individuals is therefore ever increasing. While in the U.S.A. rehabilitation programmes for TBI patients are well established, in the U.K. the provision of such services is patchy and varies widely in different localities. The belated response to the rehabilitation needs of this group of individuals in the U.K. has coincided with an increased awareness of the economic efficiency of health care provision. This paper critically reviews published studies looking at the economics of rehabilitation services for brain injured patients. No studies in the U.K. were identified and all the sources discussed are from the U.S.A. The methodological guidelines underlying economic appraisal of health care are summarised and the studies assessed to determine the extent to which they fulfil these guidelines. The paper concludes that most studies purporting to provide evidence of cost-effectiveness did not include appropriate data, nor followed the methodological guidelines allowing such claims to be made. Some recommendations for future research are presented. PMID- 9225418 TI - Communicating with the people about HIV infection risk as a basis for planning interventions: lessons from the Kagera Region of Tanzania. AB - In order to deepen the understanding of risk factors associated with HIV infection in the Kagera region of Tanzania and to investigate the potentials of communicating with the people in planning for interventions, two studies were performed in the districts of Bukoba Urban, Bukoba Rural and Muleba in 1989. The HIV prevalence of these areas ranged between 4.5% and 24.2% according to the prevalence study performed earlier in 1987. The studies involved the community in ward meetings on the one hand, and previously studied individuals on the other hand. The studies aimed both at conveying to the people the results of a previously performed study and at collecting new data using a combination of quantitative and qualitative methods in order to better understand the associated risk factors, perceived or real, and what suggestions the community could offer for reducing HIV transmission in the region. From the initial study, awareness about AIDS was found to be universal. Change of sexual partners and infection with syphilis were found to be the major risk factors for HIV-I infection. From the ward meetings people suggested a variety of solutions for interventions which we have categorized as either "hard" or "soft". The "hard" solutions involved suggestions such as isolation, imprisonment, castration and killing of AIDS victims, while the "soft" solutions involved sympathetic handling of the sick and educating the people about the modes of transmission and how best to prevent infection. There was a greater tendency for the low HIV prevalence rural communities to suggest the "hard" solutions than the high HIV prevalence urban ones which tended to suggest the "soft" solutions. However, with the changing dynamics of HIV infection in the region towards higher HIV prevalence in rural areas, it is likely that the "soft" solutions will gain acceptance and become adopted for interventions throughout the region. The information obtained from these studies has provided lessons that can be used for rational counselling as well as for guiding the implementation of IEC activities geared at interventions. It is also suggested that there should be further research into new strategies or their combinations which could be crucial in prevention such as those of community participation, empowerment of women and solidarity in AIDS intervention work. PMID- 9225419 TI - The effect of spatial definition on the allocation of clients to screening clinics. AB - We compared four strategies for inviting 91,456 women aged 50-69 years to one of six clinics for mammography screening and 40,142 men aged 60-79 years to one of 10 clinics for abdominal aortic aneurysm (AAA) screening. The strategies were invitation to the clinic nearest to the client and invitation to the clinic nearest to the client's area of residence defined by census small area, postcode and local government area. For each strategy we calculated the expected demand at each clinic and the travel distances for clients. We found that when women were allocated to mammography clinics on the basis of the local government area instead of their individual address, expected demand at one clinic increased by 60%, and 19% of clients were invited to attend a more remote clinic, entailing 99,000 km of additional travel. Similar results were obtained for men allocated to AAA clinics by their postcode of residence instead of their individual address: 55% difference in expected demand, 13% to a more remote clinic and 60,000 km of extra travel. Allocation on the basis of small areas did not show such great differences, except for travel distance, which was about 5% higher for each clinic type. We recommend that allocation of clients to screening clinics be made according to residential address, that assessment of the location of clinics be based on distances between residences and nearest clinic, but that planning new locations for clinics be aided with spatial analysis tools using small area demographic and social data. PMID- 9225420 TI - Effect of cobalt supplementation on serum vitamin B12 levels, weight gain and survival rate in lambs grazing cobalt-deficient pastures. AB - The effect of cobalt supplementation on serum vitamin B12, growth rate and survival rate was measured in controlled field experiments with Texel twin lambs of the same sex, grazing cobalt-deficient pastures. The non-supplemented lambs had lower serum vitamin B12 concentrations than their supplemented brothers or sisters. During the experiments more lambs died in the non-supplemented than in the supplemented group. At the end of the experiments supplemented lambs weighed (mean live weight) 7.2, 9.5, and 11.0 kg more than non-supplemented lambs in 1991, 1992, and 1993, respectively. Sex-related differences in weight gain and survival rate were observed. PMID- 9225421 TI - Estimating the rate of pseudorabies virus introduction into pig-finishing herds at regional level. AB - From February to June 1995, 5-12 blood samples were collected in each Dutch pig herd and tested for antibodies against pseudorabies virus (PRV). The percentage of PRV-seropositive pig-finishing herds in three regions with more than 1,000 pigs/km2 (regions 2, 4 and 7) was 6% (region 2), 12% (region 7), and 25% (region 4). The percentage of PRV-seropositive pig-finishing herds in five regions with fewer than 1,000 pigs/km2 (regions 1, 3, 5, 6, and 8) was 3% (region 1), 9% (region 3), 6% (region 5), 0% (region 6), and 4% (region 8). The small sample size allows only the detection of major outbreaks and the percentages of PRV seropositive herds therefore under-estimate the actual virus circulation in the regions. The fraction of PRV introductions that will result in a major outbreak depends on the herd immunity and thus on the vaccination programme of the herds. By combining for each herd the occurrence or absence of a major outbreak with the herd immunity induced by the vaccination programme, we estimated the average rate at which PRV was introduced into finishing herds in the eight regions. The average number of PRV introductions per finishing herd per finishing period (16 weeks) in the pig-dense regions was estimated at 0.20 in region 2, 0.83 in region 4, and 0.48 in region 7. In the less densely populated regions this rate was estimated at 0.08 in region 1, 0.34 in region 3, 0.17 in region 5, 0.00 in region 6, and 0.09 in region 8. The eight regions could be classified into four areas with a statistically different (P < 0.05 in Mann Whithey U test) rate of PRV introduction: i) regions 1, 6, and 8; ii) regions 2 and 5; iii) regions 3 and 7; and iv) region 4. PMID- 9225422 TI - The effect of intravenous or subcutaneous administration of meglumine antimonate (Glucantime) in dogs with leishmaniasis. A randomized clinical trial. AB - The efficacy of i.v. versus s.c. administration of Glucantime (100 mg/kg of body weight/day) was studied in 41 dogs with leishmaniasis without serious renal insufficiency. Remission was obtained in 35 dogs (85.4%) after 3 to 6 weeks of treatment but there was a relapse within 1 year in 26 dogs (74.3%). The median period of remission was 6 months. Cross-over therapy resulted in remission in 17 of 20 dogs. The percentage of remission after initial and cross-over therapy, the median relapse free period, and survival did not differ significantly between the two groups. There were very few complications and most were of minor clinical importance. Thrombophlebitis developed in one dog after i.v. injection. In dogs with leishmaniasis without serious renal insufficiency, there is a 75% probability of survival for more than 4 years following treatment with Glucantime for 3 to 6 weeks, with additional treatment when relapses occur. PMID- 9225423 TI - Prevalence of intestinal nematodes of dogs and cats in The Netherlands. AB - Faecal samples from 272 dogs and 236 cats from Dutch households were examined for nematode eggs. Toxocara eggs were found in 8 dogs (2.9%) and 11 cats (4.7%). Toxascaris eggs were found in 1 dog (0.4%) and Trichuris eggs in 2 dogs (0.7%). Examination of faeces from 56 stray cats revealed Toxocara in 12 cases (21%) and Toxascaris eggs in 3 cases (5.4%). No hookworm eggs were found. The percentage of positive samples was significantly higher in young animals than in older animals. Toxocara eggs were found significantly more frequently in stray cats than in cats kept in households. PMID- 9225424 TI - Urinary corticoid/creatinine ratios in the differentiation between pituitary dependent hyperadrenocorticism and hyperadrenocorticism due to adrenocortical tumour in the dog. AB - In a study on the differentiation between pituitary-dependent hyperadrenocorticism (PDH) and hyperadrenocorticism due to adrenocortical tumour (AT), two questions were addressed: 1. Do basal urinary corticoid/creatinine (c/c) ratios have any value in this respect, and 2. what is the reference percentage suppression of the urinary c/c ratios in the high-dose dexamethasone suppression test? Data obtained from 160 dogs with hyperadrenocorticism were analysed. In 49 dogs the diagnosis AT was confirmed by the finding of plasma ACTH concentrations < 40 ng/l, by visualisation of the tumour by ultrasonography and/or computed tomography, and by histological examination of the adrenal tissue obtained at surgery or autopsy. Among the 111 dogs with PDH, there were 31 animals with resistance to dexamethasone suppression, i.e., suppression < 50%. The basal urinary c/c ratios of dogs with PDH and AT did not differ significantly, although urinary c/c ratios > 100 x 10(-6) almost exclusively occurred in association with PDH. Among the dogs with hyperadrenocorticism, the positive predictive value of a basal urinary c/c ratio > 100 x 10(-6) for the diagnosis of PDH was 0.90 (95% CI: 0.74-0.98). Of the 49 dogs with AT, 34 had a urinary c/c ratio after dexamethasone administration higher than the basal urinary c/c ratio. The maximum suppression of the basal urinary c/c ratio in dogs with AT was 43.7%. It is concluded that in dogs with hyperadrenocorticism basal urinary c/c ratios only have predictive value in the differentiation between AT and PDH when the ratio exceeds 100 x 10(-6). The generally accepted criterion of 50% suppression by dexamethasone in the differentiation between PDH and AT is also applicable to the urinary c/c ratio. PMID- 9225425 TI - Comparison of two suture materials for intradermal skin closure in dogs. AB - The macroscopic and histological appearance of cutaneous incisions closed with polyglecaprone 25 (Monocryl) and polyglactin 910 (Vicryl) suture materials were compared in four dogs. Polyglecaprone 25 compared favourably to polyglactin 910 suture material for closure of canine skin incisions, and was associated with significantly less tissue reaction in the early phases of the healing process than was polyglactin 910 suture material. This difference was not present at later evaluation times. PMID- 9225426 TI - Long-term success rate of resin-bonded metal crowns on the canine teeth of working dogs. AB - In this clinical study, 19 full metal crown restorations of canine teeth were placed in seven working dogs. Thirteen canine teeth were severely abraded with no involvement of the pulp cavities; six fractured canine teeth were endodontically treated. At least 1/3 of the coronal part of the canine tooth was available for a supragingivally performed, minimal tooth crown preparation. An adhesive technique to bond the electrolytically etched crown (an alloy of cobalt-chrome-molybdenum) to the tooth was used. The metal crowns, slightly shorter and with a rounder tip than the original tooth, were bounded to the enamel and dentine by using a resin luting cement. Posts or post-and-core techniques were not used. Mean follow-up period was 32 months (range 24-52 months), at which stage 17 crowns were found to be intact and functional. Two crowns were lost as a result of trauma resulting in a fracture of the tooth below the crown. PMID- 9225427 TI - Monoclonal gammopathy in a Dutch warmblood mare. AB - A 15-year-old Dutch warmblood mare was presented because of lethargy, which had been present for several weeks, and severe anaemia. Total protein was high and serum electrophoresis revealed a monoclonal peak in the alpha-2 region. Monoclonal immunoglobulin, IgG(T), was detected by immuno-electrophoresis in serum and urine. Postmortem examination revealed a relatively large number of plasmacytoid cells in the bone marrow and a monotonous population of plasmacytoid cells in the spleen. These findings are suggestive of a plasma cell myeloma. PMID- 9225428 TI - Different seasonal occurrence of anaplasmosis outbreaks in beef and diary cattle in an area of Argentina free of Boophilus microplus ticks. AB - The seasonal occurrence of anaplasmosis (Anaplasma marginale) outbreaks in dairy and beef cattle was analysed for a region of Argentina (29 degrees to 31 degrees S and 58 degrees to 62 degrees W) that is free of Boophilus microplus ticks, using data collected from December 1978 to November 1995. The outbreaks were confirmed by inspection of blood smears obtained from sick or dead cattle. A total of 94 outbreaks were confirmed by inspection of blood smears obtained from sick or dead cattle. A total of 94 outbreaks were identified: 48 in beef cattle and 46 in dairy herds. The proportional seasonal distribution of outbreaks was different in beef and dairy cattle (chi-square = 15.08, P < 0.01). While no seasonal pattern of anaplasmosis outbreaks was found in dairy cattle, there was a concentration of outbreaks during the summer months (54% of the total) in beef cattle. Rural practices that are carried out more frequently on dairy than on beef cattle may have favoured iatrogenic transmission of A. marginale in the milk production system. PMID- 9225429 TI - Progressive neuronopathy in two Cairn terrier litter mates. AB - Clinical, histopathological, and EM findings are described for two Cairn terrier litter mates, an 18-months-old male and an 11-month-old female with progressive neuronopathy. The initial clinical signs were characterized by hind limb weakness and ataxia, which deteriorated with exercise. These signs progressed over several months to tetraparesis. Pathological examination revealed extensive chromatolytic degeneration of neurons and moderate secondary Wallerian-type degeneration in the spinal cord and brain stem. Progressive neuronopathy can be differentiated clinically from globoid cell leukodystrophy, another progressive neurological disorder in Cairn terriers, by the exercise-induced deterioration of the neurological signs. Progressive neuronopathy occurs only in Cairn terriers and because of the similarity in age of onset and the occurrence in one litter, an inherited disease is suspected. PMID- 9225430 TI - The effect of discontinuation of postmilking teat disinfection in low somatic cell count herds. I. Incidence of clinical mastitis. AB - Results are described of a split-udder trial on the effect of discontinuation of postmilking teat disinfection on the incidence of clinical mastitis in seven dairy herds with a low bulk milk somatic cell count and a high incidence of clinical mastitis. Overall incidence of clinical mastitis was non-significantly lower (18%), whereas the incidence of the most prevalent pathogen associated with clinical mastitis, Escherichia coli, was significantly lower in quarters for which postmilking teat disinfection was discontinued. We concluded that discontinuation of postmilking teat disinfection may decrease the incidence of clinical Escherichia coli mastitis in herds for which standard mastitis prevention measures are executed adequately, bulk milk somatic cell count is low, and incidence of clinical mastitis is high. However, because an increase in intramammary infections with contagious pathogens may occur, care is recommended when advising discontinuation of postmilking teat disinfection. PMID- 9225431 TI - The effect of discontinuation of postmilking teat disinfection in low somatic cell count herds. II. Dynamics of intramammary infections. AB - Results of a 20 month split-udder trial on the effect of discontinuation of postmilking teat disinfection on intramammary infections (IMI) with major and minor pathogens in seven dairy herds with a low somatic cell count are described. The incidence of Escherichia coli IMI was found to be significantly lower, whereas the incidence of IMI with Staphylococcus aureus and minor pathogens was significantly higher in quarters for which postmilking teat disinfection was discontinued than in disinfected quarters. It was concluded that discontinuation of postmilking teat disinfection decreased the incidence of E. coli IMI, accompanied by a, from a practical point of view, acceptable rise in somatic cell count. However, the possible increase in the incidence of S. aureus IMI calls for careful monitoring of the dynamics of IMI with contagious pathogens, when postmilking teat disinfection is discontinued in an attempt to reduce E. coli mastitis. PMID- 9225433 TI - Microphthalmia, brachygnathia superior, and palatocheiloschisis in a foal associated with griseofulvin administration to the mare during early pregnancy. AB - An 18 year old Friesian mare was treated with griseofulvin for dermatomycosis in the second month of pregnancy. Pregnancy was uneventful and after 331 days a male foal was born. The foal showed bilateral microphthalmia, severe brachygnathia superior, and palatocheiloschisis. The lesions were incompatible with life and the animal was euthanized. As similar lesions have been described in other species associated with griseofulvin administration during pregnancy, and the development of the eyes and facial bones in the horse occurs in the second month of pregnancy, the lesions most likely can be attributed to griseofulvin administration. PMID- 9225432 TI - Characterization of muscarinic receptors in equine tracheal smooth muscle in vitro. AB - This study was undertaken to assess the importance of muscarinic receptor subtypes in equine airway disease. Smooth muscle strips from the mid-cervical portion of the trachea of horses were placed in tissue baths and isometric contractile force was measured. Active force was measured in response to metacholine and the selective muscarinic receptor agonists McN-A-343 (M1 selective) and pilocarpine (M2-selective) in cumulative concentrations (10(-9)M through 10(-3)M), with and without preincubation with three or four concentrations of the selective muscarinic receptor antagonists pirenzepine (M1 selective), methoctramine (M2-selective), and 4-DAMP (M3-selective). The tissues contracted in response to all muscarinic agonists. The maximum responses (mean +/ sem) were 86.7 +/- 6.2 g for metacholine, 27.1 +/- 2.5 g for McN-A-343 and 37.6 +/- 3.5 g for pilocarpine. Preincubation with the selective muscarinic receptor antagonists resulted in dose-dependent rightward shifts of the concentration effect curves for metacholine. pA2 values (means +/- sem) were 8.88 +/- 0.30 for 4-DAMP, 6.53 +/- 0.38 for methoctramine, and 6.72 +/- 0.31 for pirenzepine. Preincubation with 10(-7) M 4-DAMP resulted in a rightward shift of the concentration-effect curves for McN-A-343 and pilocarpine. These results indicate that the most important muscarinic receptor mediating contraction of equine tracheal smooth muscle is of the M3-type. Therefore relatively low concentrations of a M3-selective muscarinic receptor antagonist will inhibit acetylcholine induced contraction of equine airway smooth muscle. PMID- 9225434 TI - An attempt to control lungworm disease and parasitic gastroenteritis on commercial dairy farms by the use of an adaptation of Michel's dose-and-move system. AB - On seven dairy farms an attempt was made to control lungworm disease in calves by turnout on a pasture grazed earlier by cows, followed by a move to aftermath and ivermectin treatment 2 months later. Transmission of lungworm was observed on all farms. Lungworm disease occurred on four farms at treatment. Coughing re-occurred on three of these farms in some animals 2 months later. Owing to poor performance between turnout and treatment, weight gain was below the norm on the farm with the highest infections and most severe respiratory signs. On the other farms respiratory signs did not result in poor weight gain. Gastrointestinal nematode infections remained low on all farms. The conclusion is that this dose and move scheme cannot be recommended for the control of lungworm. PMID- 9225435 TI - Intramuscular bioavailability of ketoprofen lysine salt in horses. AB - Lysine salts are often used in human pharmaceuticals to increase the solubility and absorption of acidic drugs when these are administered parenterally. In this study the intramuscular bioavailability of ketoprofen administered as the lysine salt was evaluated in horses (n = 5) treated intravenously and intramuscularly (2.2 mg/kg active substance) in a cross-over study. The absorption rate of ketoprofen administered as the lysine salt was rather low: the mean residence time increased from 31.7 min after IV injection to 128.9 min (after IM injection), and the bioavailability was high (mean 92.4%). The calculated steady state plasma concentrations of ketoprofen during multiple dosage were much higher after intramuscular (0.106 g/ml) than after intravenous (0.066 microgram/ml) administration. Intramuscular injections of the ketoprofen lysine salt can therefore be given to horses, which are particularly prone to develop soft tissue reactions, since use of the lysine salt markedly reduced local irritation at the injection site. PMID- 9225436 TI - A case of autochthonous canine leishmaniasis in The Netherlands. AB - The transmission of visceral leishmaniasis (VL) in the absence of its natural vector, the sandfly, is considered exceptional. This report describes VL in a 12 month-old dog which had never been in an area in which VL is endemic but was born in the Netherlands from a bitch that had been infected in Spain. Although the mode of transmission, via the placenta or otherwise, is unknown, it can be concluded that bitches with VL can be a source of infection for their pups, even in a sandfly-free non-endemic area. The dog was successfully treated with allopurinol. PMID- 9225437 TI - Acute colitis in adult horses. A review with emphasis on aetiology and pathogenesis. AB - This review article describes the different aetiological agents known or suspected to cause colitis in the adult horse, namely Salmonella spp., Clostridium spp., Ehrlichia risticii, Cyathostomes, fungi, various antibiotics, drugs, and toxins, with emphasis on their mechanism of action. For each of the infectious agents, diagnostic procedures are indicated. The effects of endotoxin can be important in all forms of equine colitis. PMID- 9225438 TI - Primary hyperparathyroidism in two cats. AB - Primary hyperparathyroidism (PHP) is an infrequently diagnosed disorder in cats. In this report the signs and symptoms of two cats with hypercalcaemia due to PHP are described, together with diagnostic approach, results of treatment, and immunohistochemical findings. A 9-year-old and a 13-year-old neutered male domestic shorthair cat were presented with signs of lethargy, anorexia, and vomiting. Both cats had persistent hypercalcaemia and normo- to hypophosphataemia. Cytological examination of a fine-needle aspiration biopsy sample of a palpable cervical mass revealed groups of benign glandular-epithelial cells in one cat. In the other cat no cervical mass was palpable. In this cat plasma parathyroid hormone (PTH) levels were measured repeatedly and these values exceeded the maximum reference value on two occasions. Following exclusion of other causes of hypercalcaemia both cats were subjected to neck surgery and in both a solitary parathyroid adenoma was removed. The adenomas contained an abundance of PTH, as demonstrated by immunohistochemical techniques. Plasma calcium and phosphate concentrations returned to within, reference ranges postoperatively. Recovery was uncomplicated and there were no signs of recurrence on follow-up examinations. PMID- 9225439 TI - Surgical treatment and complications of a urethral obstruction in a cat: a case study. AB - A 4-year-old Burmese cat had undergone two different urethrostomy procedures to resolve feline urethral obstruction. Both methods failed because of incorrect surgical techniques. The reasons for failure, the secondary complications, and the surgical correction of the previously performed urethrostomies are described. PMID- 9225440 TI - Factors influencing complications during caesarean section on the standing cow. AB - The complications occurring during Caesarean section are reviewed, and then an investigation of the factors influencing three important complications, namely recumbency of the animal, increased contractility of the uterus, and difficulties with exteriorization of the pregnant uterine born, is described. Two epidemiological and statistical methods were used. The parity, the type of animal, the use of sedatives, and difficulties with the exteriorization of the pregnant horn had, with increasing significance, an independent influence on recumbency of the cow. Attempts to extract the calf was the only factor that significantly increased uterine contractility. The surgeon, the parity, the increased uterine contractility, the position of the calf, and the presence of adhesions were associated with increasing significance, with difficult exteriorization of the pregnant horn. It is very important to know the factors that influence the occurrence of complications during surgery, in order to take some precautions to minimize the complications after the operation. PMID- 9225441 TI - Riboprinting: a tool for the study of genetic diversity in microorganisms. AB - Classical morphology-based methods of taxonomic and phylogenetic analysis are inadequate in many groups of structurally simple eukaryotes. Molecular methods can generate data independently of the complexity of the organisms' morphology. Riboprinting is one such technique, and involves restriction enzyme analysis of polymerase chain reaction amplified small subunit ribosomal RNA genes. The utility of the method is illustrated with examples from several genera of intestinal and bloodstream parasites. Among the applications of riboprinting are the detection of cryptic genetic variation within species, organism misidentifications and culture mix-ups, independent verification of DNA sequences, and the rapid generation of data useful in phylogenetic analyses. PMID- 9225442 TI - Phagosomal proteins of Dictyostelium discoideum. AB - In recognizing food particles. Dictyostelium cell-surface molecules initiate cytoskeletal rearrangements that result in phagosome formation. After feeding D. discoideum cells latex beads, early phagosomes were isolated on sucrose step gradients. Protein analyses of these vesicles showed that they contained glycoproteins and surface-labeled species corresponding to integral plasma membrane proteins. Cytoskeletal proteins also were associated with phagosomes, including myosin II, actin and a 30 kDa-actin bundling protein. As seen by the acridine orange fluorescence of vesicles containing bacteria, phagosomes were acidified rapidly by a vacuolar H(+)-ATPase that was detected by immunoblotting. Except for the loss of cytoskeletal proteins, few other changes over time were noted in the protein profiles of phagosomes, suggesting that phagosome maturation was incomplete. The indigestibility of the beads possibly inhibited further endocytic processing, which has been observed by others. Since nascent phagosomes contained molecules of both the cytoskeleton and plasma membrane, they will be useful in studies aimed at identifying specific protein associations occurring between membrane proteins and the cytoskeleton during phagocytosis. PMID- 9225443 TI - In vitro cultivation of the vascular phase of Sarcocystis singaporensis. AB - To establish an in vitro culture system for the precystic phase of Sarcocystis singaporensis, we initially tested various excysting fluids for sporocysts. An excysting fluid containing 2.5% bovine taurocholate and 10% bile of the specific intermediate host, Rattus norvegicus, in RPMI medium was the most suitable resulting in excystation of 80% of the sporozoites. Subsequently, we identified brain endothelial cells and pneumonocytes of the rat to promote growth of sporozoites to schizonts. Hepatoma, fibroblastic, or myoblastic cells were not suitable for the parasite's development. First-generation schizonts were seen at days 3-10 postinoculation (PI); a distinct second peak of schizogonic development only occurred in endothelial cells at days 14-18 PI. First-generation schizonts were 26.0 (+/- 3.8) microns in diameter and contained 32-50 merozoites, second generation schizonts measured 34.4 (+/- 10.6) microns and contained 54-72 merozoites. Merozoite yield at large-scale culture conditions (75 cm2 flasks) using pneumonocytes as host cells was relatively low. Ultrastructurally, sporozoites and merozoites were quite similar to corresponding stages of other Sarcocystis species. With regard to host cell specificity and developmental kinetics, in vitro cultivation showed close similarities to the situation in vivo. PMID- 9225444 TI - Fluorescently-labeled oligonucleotide probes can be used to identify protistan food vacuole contents. AB - In situ hybridization using fluorescent oligonucleotide probes complementary to unique regions of 16S rRNA molecules provides a way of identifying the food vacuole contents of bactivorous protists. Laboratory experiments with Tetrahymena showed rRNAs in food vacuoles are degraded slowly enough to permit their use as hybridization targets for such probes. A probe specific for a hypervariable region of the small subunit rRNA of an unnamed proteobacterium abundant in a local lake was then synthesized. It was used to probe the food vacuoles of the ciliates present in fixed water samples collected from the same lake. The vacuoles of several filter-feeding ciliates bound the probe, indicating that such probes can be used to identify the food vacuole contents of ciliates collected from natural samples. PMID- 9225445 TI - Ribosomal RNA analysis indicates a benthic pennate diatom ancestry for the endosymbionts of the dinoflagellates Peridinium foliaceum and Peridinium balticum (Pyrrhophyta). AB - The establishment of chloroplasts as cellular organelles in the dinoflagellate, heterokont (stramenopile), haptophyte, and cryptophyte algae is widely accepted to have been the result of secondary endosymbiotic events, that is, the uptake of a photosynthetic eukaryote by a phagotrophic eukaryote. However, the circumstances that promote such associations between two phylogenetically distinct organisms and result in the integration of their genomes to form a single functional photosynthetic cell is unclear. The dinoflagellates Peridinium foliaceum and Peridinium balticum are unusual in that each contains a membrane bound eukaryotic heterokont endosymbiont. These symbioses have been interpreted, through data derived from ultrastructural and biochemical investigations, to represent an intermediate stage of secondary endosymbiotic chloroplast acquisition. In this study we have examined the phylogenetic origin of the P. foliaceum and P. balticum heterokont endosymbionts through analysis of their nuclear small subunit ribosomal RNA genes. Our analyses clearly demonstrate both endosymbionts are pennate diatoms belonging to the family Bacillariaceae. Since members of the Bacillariaceae are usually benthic, living on shallow marine sediments, the manner in which establishment of a symbiosis between a planktonic flagellated dinoflagellate and a bottom-dwelling diatom is discussed. In particular, specific environmentally-associated life strategy stages of the host and symbiont, coupled with diatom food preferences by the dinoflagellate, may have been vital to the formation of this association. PMID- 9225446 TI - Effect of platelet-activating factor on the process of cellular differentiation of Herpetomonas muscarum muscarum. AB - The effects of platelet-activating factor (PAF), at doses ranging from 10(-6) M to 10(-10) M, on cell growth and on cell differentiation of Herpetomonas muscarum muscarum were investigated. Cell differentiation was evaluated by both light and electron microscopy. At the concentrations used, PAF did not interfere with the protozoan growth. However, parasites grown in the presence of PAF (10(-6) M) were significantly more differentiated than those grown in the absence of PAF, since the first day of culture. On the first two days of culture, PAF doses ranging from 10(-10) M to 10(-7) M, did not significantly interfere with the differentiation of these parasites, although after the third day of culture, all PAF doses used significantly increased the protozoan differentiation. Specific PAF receptor antagonists totally abrogated (WEB 2086 and WEB 2170) or significantly decreased (BN 52021) PAF effect on cell differentiation. These findings indicate PAF triggers the process of cell differentiation in Herpetomonas muscarum muscarum and suggest these parasites have receptors for PAF. PMID- 9225447 TI - Distribution of epitopes of Trypanosoma cruzi amastigotes during the intracellular life cycle within mammalian cells. AB - In this study we have examined the distribution of epitopes defined by monoclonal antibodies raised against Trypanosoma cruzi amastigotes during the intracellular life cycle of the parasite. We have raised monoclonal antibodies towards amastigote forms and performed preliminary immunochemical characterization of their reactivities. MAB 1D9, 3G8, 2B7, 3B9, and 4B9 react with carbohydrate epitopes of the parasite major surface glycoprotein--Ssp-4 defined by MAB 2C2 [5]; MAB 4B5 reacts with a noncarbohydrate epitope in all developmental stages of the parasite, and MAB 3B2 also detects a noncarbohydrate epitope preferentially in T. cruzi flagellated forms. Vero cells infected with tissue culture-derived trypomastigotes of clone D11 (G strain) were fixed at different times during the intracellular proliferation of parasites, and processed for immuno-electron microscopy and confocal immunofluorescence with the different monoclonal antibodies. We observed that while the surface distribution of MAB 2C2 and 4B9 epitopes was uniform throughout the cycle, MAB 1D9, 3G8, and 2B7 reacted with cytoplasmic membrane-bound compartments of the amastigotes. MAB 3B9 displayed a unique surface dentate pattern in some amastigotes. MAB 4B5 recognized a curved shaped structure at the flagellar pocket region in some intracellular amastigotes and localized to the membrane in dividing forms. In intracellular trypomastigotes, MAB 4B5 also displayed a punctate pattern near the flagellar pocket. PMID- 9225448 TI - Rapid methylation of cell proteins and lipids in Trypanosoma brucei. AB - The fate of the [methyl-14C] group of S-adenosylmethionine (AdoMet) in bloodstream forms of Trypanosoma brucei brucei, was studied. Trypanosomes were incubated with either [methyl-14C]methionine, [U-14C]methionine, S-[methyl 14C]AdoMet or [33S]methionine and incorporation into the total TCA precipitable fractions was followed. Incorporation of label into protein through methylation was estimated by comparing molar incorporation of [methyl-14C] and [U 14C]methionine to [35S]methionine. After 4-h incubation with [U-14C]methionine, [methyl-14C]methionine or [35S]methionine, cells incorporated label at mean rates of 2,880 pmol, 1,305 pmol and 296 pmol per mg total cellular protein, respectively. Cells incubated with [U-14C] or [methyl-14C]methionine in the presence of cycloheximide (50 micrograms/ml) for four hours incorporated label eight- and twofold more rapidly, respectively, than cells incubated with [35S]methionine and cycloheximide. [Methyl-14C] and [U-14C]methionine incorporation were > 85% decreased by co-incubation with unlabeled AdoMet (1 mM). The level of protein methylation remaining after 4-h treatment with cycloheximide was also inhibited with unlabeled AdoMet. The acid precipitable label from [U 14C]methionine incorporation was not appreciably hydrolyzed by DNAse or RNAse treatment but was 95% solubilized by proteinase K. [U-14C]methionine incorporated into the TCA precipitable fraction was susceptible to alkaline borate treatment, indicating that much of this label (55%) was incorporated as carboxymethyl groups. The rate of total lipid methylation was found to be 1.5 times that of protein methylation by incubating cells with [U-14C]methionine for six hours and differential extraction of the TCA lysate. These studies show T. b. brucei maintains rapid lipid and protein methylation, confirming previous studies demonstrating rapid conversion of methionine to AdoMet and subsequent production of post-methylation products of AdoMet in African trypanosomes. PMID- 9225450 TI - The plaque matrix (PQM) and tubules at the surface of intramuscular parasite, Trachipleistophora hominis. AB - Surface plaque matrix (PQM) and a tubular arrangement of filaments border Trachipleistophora hominis parasites during growth within host muscle. The PQM at the parasite surface forms a network of processes which can be associated with filamentous tubules. Peroxidase tracer delineated the PQM and showed apparent connections with the tubules. The tubules at the interface of T. hominis-infected cells are structurally similar to the extrasporular tubules of the microsporidian, Ameson michaelis. The extrasporular tubules of A. michaelis and the proteins from T. hominis-infected muscle reacted to keratin antibodies, K8.13, K4 and K13. Conversely, antibodies produced to T. hominis-infected muscle, reacted with the extrasporular tubular proteins of A. michaelis. The PQM and tubular elements are thought to play an important role in affecting molecular traffic between the host and parasite. PMID- 9225449 TI - Effects of carboxylmethylation and polyamine synthesis inhibitors on methylation of Trypanosoma brucei cellular proteins and lipids. AB - The fate of methionine in eukaryotic cells is divided between protein synthesis and the branched pathway encompassing polyamine synthesis, methylation of proteins and lipids, and transsulphuration reactions. Aside from protein synthesis, the first step to all other uses of methionine is conversion to S adenosylmethionine. Blockade of polyamine synthesis in African trypanosomes by the ornithine decarboxylase inhibitor DL-alpha-difluoromethylornithine (Ornidyl, DFMO) the AdoMet decarboxylase inhibitor 5'-[[(Z)-4-amino-2-butenyl]-methylamino] 5'-deoxyadenosine or the protein methylase inhibitor sinefungin induces dramatic increases in intracellular AdoMet. In a previous study, distribution and pool sizes of [35S] or [U-14C]methionine were followed in bloodform trypanosomes as incorporation into the total TCA precipitable fractions. In the present study, the effects of pretreatment with DFMO (1 mM), MDL 73811 (1 microM) and sinefugin (2 nM) on [35S] and [U-14C]methionine incorporation were studied in blood forms. DFMO or MDL 73811 pretreatment increased protein methylation 1.5-fold through incorporation of [U14C]methionine, while sinefungin caused a 40% reduction of incorporation. The increases in incorporation of [U-14C]methionine due to DFMO and MDL 73811 were reduced 40% to 70% by including cold AdoMet (1 mM) in the incubation medium, an indication of AdoMet transport by bloodform trypanosomes and the utilization of [U-14C]methionine as AdoMet. Exogenous AdoMet had no effect on [35S]methionine incorporation. The agents studied are curative for African trypanosomiasis infections, either clinically (DFMO) or in model infections (MDL 73811, sinefungin) and thus highlight interference with AdoMet metabolism and methylation reactions as biochemical consequences of these agents. PMID- 9225451 TI - The role of the nucleus isthmi in respiratory pattern formation in bullfrogs. AB - The nucleus isthmi (NI) is a mesencephalic structure of the amphibian brain located between the roof of the midbrain and the cerebellum. From a neuroanatomical perspective, the NI can be compared with the pons which, in mammals, contributes to the control of breathing pattern. This study tested the hypothesis that the NI plays a critical role in breathing pattern formation in the bullfrog. More specifically, we postulated that this nucleus was the site responsible for clustering breaths into distinct episodes of breathing. This hypothesis was tested by comparing the respiratory motor output of decerebrate, paralyzed and artificially ventilated bullfrogs before and after bilateral lesions of the NI by pressure microinjections of lidocaine or kainic acid (KA) into this area. Bilateral microinjections of lidocaine or KA into the NI transformed the breathing pattern from episodic (many breaths per episode) to one of evenly spaced single breaths, without affecting the amplitude of the fictive breaths. These changes in breathing pattern were associated with an overall decrease in breathing frequency and a reduction in CO2-chemosensitivity. Breathing episodes of more than one breath reappeared during hypercarbia (3.5% CO2 in air) after KA lesioning. Bilateral lesions to the NI did not affect the changes in the timing or the amplitude of the respiratory-related bursts elicited by pulmonary stretch receptor feedback, indicating that mechanoreflexes do not require NI input. We conclude that the NI is not responsible for the genesis of breathing episodes, but provides a tonic excitatory input to respiratory centers in the lower brainstem. The NI also plays an important role in either CO2 chemodetection or, more probably, integration of CO2 chemoreceptor information. This, in turn, contributes to the production of episodes of more than one breath. PMID- 9225452 TI - Evidence for a persistent Na+ conductance in neurons of the gastric mill rhythm generator of spiny lobsters. AB - Evidence for a persistent Na+ conductance was obtained in identified motor neurons of the gastric mill network in the stomatogastric ganglion of the spiny lobster Panulirus interruptus. The cells studied were the lateral gastric and lateral posterior gastric motor neurons, which in vivo control chewing movements of the lateral teeth of the gastric mill. We examined basic cellular properties in the quiescent network of the isolated stomatogastric ganglion. In current clamp recordings, we found two types of evidence for a persistent Na+ conductance. First, tetrodotoxin-sensitive inward rectification occurred during depolarization from rest to spike threshold. Second, 5 mmol l-1 tetraethylammonium (a K+ channel blocker) induced plateau potentials that persisted in the presence of Mn2+ or a low [Ca2+]0 but were blocked by a low [Na+]0 or 100 nmol l-1 tetrodotoxin. The plateau potentials could drive trains of fast spikes in the motor axon and strong transmitter release at central output synapses within the ganglion. This conductance probably corresponds to the persistent Na+ current, INaP, described in cultured stomatogastric neurons and in neurons from several other preparations. During normal neuronal activity, it may contribute to the prolonged plateau depolarizations and long spike trains typical of motor neuronal activity during gastric rhythm generation. Persistent inward currents of this type are likely to be important in neurons that must fire prolonged bursts in cycle after cycle of rhythmical activity. PMID- 9225454 TI - To treat or not to treat: the case of tuberculosis. AB - Incomplete treatment of patients with infectious tuberculosis (TB) may not only lead to relapse but also to the development of antibiotic resistant TB-one of the most serious health problems facing society today. In this article, we formulate one-strain and two-strain TB models to determine possible mechanisms that may allow for the survival and spread of naturally resistant strains of TB as well as antibiotic-generated resistant strains of TB. Analysis of our models shows that non-antibiotic co-existence is possible but rare for naturally resistant strains while co-existence is almost the rule for strains that result from the lack of compliance with antibiotic treatment by TB infected individuals. PMID- 9225453 TI - Physiological mechanisms of evolved desiccation resistance in Drosophila melanogaster. AB - We investigated physiological characters associated with water balance in laboratory populations of Drosophila melanogaster selected for resistance to desiccating conditions for over 100 generations. Five replicate, outbred, desiccation-selected (D) populations were compared with their control (C) populations. Water loss rates of female D flies were approximately 40% lower than those of C females. Although excretory water loss was reduced in desiccation selected flies, it comprised less than 10% of total water loss, indicating that the D populations have evolved reduced cuticular and/or respiratory water loss rates. Total surface lipid amounts did not differ between the C and D flies. Cuticular hydrocarbons from D flies were longer than those from C flies and melted at slightly higher temperatures, possibly contributing to reduced water loss rates. Desiccation-selected flies contained approximately 30% more bulk water than controls, as well as more glycogen. However, total metabolic water stores did not differ between the stocks owing to higher lipid levels in the C populations. The ability to tolerate water loss, as measured by water content at the time of death, did not differ between D and C flies. Thus, evolution of increased desiccation resistance has occurred by multiple physiological mechanisms, but some potential adaptive differences have not evolved. PMID- 9225455 TI - On the non-existence of an optimal migration rate. AB - We show that an optimal migration rate may not exist in a population distributed over an infinite number of individual living sites if empty sites occur. This is the case when the mean number of offspring per individual mu is finite. We make the assumption of uniform migration to other sites whose rate is determined by the parent's genotype or the offspring's genotype at a single locus in a diploid hermaphrodite population undergoing random mating. In both cases, for mu small enough, any population at fixation would go to extinction. Moreover, in the latter case, for intermediate values of mu, the only fixation state that could resist the invasion of any mutant would lead the population to extinction. These are the two conditions for the non-existence of an optimal migration rate. They become less stringent as the cost for migration expressed by a coefficient of selection 1-beta becomes larger, that is closer to 1. The results are obtained assuming that the allele at fixation is either nondominant or dominant. Although the optimal migration rate is the same in both cases when it exists, the optimality properties may differ. PMID- 9225456 TI - Clinical and mechanistic aspects of photopheresis. AB - Photopheresis is an extracorporeal form of photochemotherapy with 8 methoxypsoralen (8-MOP) and ultraviolet A (UVA) radiation. Photopheresis is used for the management of T-cell-mediated diseases, and such treatment leads to the induction of antigen-specific immune suppression directed to the pathogenic clone of T cells. Photopheresis is used to treat a wide variety of diseases--such as cutaneous T-cell lymphoma, systemic sclerosis; rheumatoid arthritis, lupus erythematosus--and is also successfully applied in the suppression of graft rejection. In addition to the clinical achievements, attention will be paid to results from animal studies. An important outcome of these studies is that photopheresis can be used to treat airway hyperreactivity. Furthermore, it was shown that the therapeutic strategy can be changed drastically: the presence of plasma during irradiation should be avoided and the amount of blood that must be treated to obtain the desired antigen-specific immunosuppression can be greatly decreased. Also, results from cellular experiments are discussed. An example of this is the increase in the major histocompatibility complex expression on the surface of cells found after treatment. The mechanism that underlies photopheresis has not yet been elucidated, but progress has been made. The following related points will be reviewed: models for investigation; and mechanistic aspects, with the emphasis on cellular biomacromolecules and on photosensitizers (drugs) other than 8-MOP. PMID- 9225457 TI - Pulsed photoacoustic spectroscopy applied to the diffusion of sunscreen chromophores in human skin: the weakly absorbent regime. AB - Pulsed photoacoustic spectroscopy was used to study the penetration of sunscreen chromophores into human skin. This study focuses on basic solutions containing single typical filter molecules, as used in current sunscreens, dissolved in mineral oil. The pulsed form of the photoacoustic technique was preferred because it provides more detailed information on the filter distribution within the different layers of human skin. A new methodology provides better insight into the diffusion process through signal analysis in the time and frequency domains, allowing for global and depth-related characterization. The penetration of the chromophore influences the response signal by inducing changes in the optical and thermal properties at different depths within the medium. The light scattering effect of titanium dioxide was demonstrated by the same technique. PMID- 9225458 TI - Sunscreens prevent local and systemic immunosuppression of contact hypersensitivity in mice exposed to solar-simulated ultraviolet radiation. AB - Ultraviolet (UV) irradiation causes the immunosuppression of contact hypersensitivity (CH) responses in animals and humans. There are conflicting reports regarding the effectiveness of sunscreens in preventing UV-induced suppression of both local-type CH (induced by the application of the contact sensitizer directly to UV-exposed skin) and systemic-type CH (induced by the application of the contact sensitizer to an unirradiated skin site 3 days after UV exposure). The purposes of this study were as follows: 1. to derive solar simulator UV dose-response curves for the induction of local and systemic CH suppression in C3H mice; 2. to establish minimum immune suppression doses (MISDs) for local and systemic CH; 3. to determine the local and systemic immune protection capacity of two commercial sunscreen lotions with labeled sun protection factors (SPFs) of 4 and 8. Dose-response curves for the induction of local and systemic CH suppression were derived by exposing groups of mice to a range of full-spectrum UV doses (0.37-21.4 kJ m-2) on two consecutive days delivered from a filtered 1000 W xenon arc lamp solar simulator. The MISDs, defined as the lowest dose tested to cause approximately 50% suppression of the normal CH response, were obtained from the dose-response curves. Although the local and systemic immunosuppression dose-response curves were not statistically different, the MISD for local suppression of CH (1.35 kJ m-2) was about fivefold lower than that for systemic CH suppression (6.76 kJ m-2). The MISD was used as the endpoint to determine sunscreen immune protection levels. Both sunscreens, applied at 2 mg cm-2, provided immune protection against the induction of local and systemic CH suppression in mice exposed to an effective UV dose of 1 MISD given through the sunscreen, i.e. 4 MISD to SPF 4 sunscreen-protected mice and 8 MISD to SPF 8 sunscreen-protected mice mounted CH responses that were significantly greater than those elicited in unprotected mice exposed to 1 MISD of solar-simulated UV radiation. The calculated immune protection factors for these sunscreens exceeded the level of protection predicted by their labeled SPFs, i.e. the local immune protection factor of both sunscreens was 15 and the systemic immune protection factors were 8 for the SPF 4 sunscreen and 15 for the SPF 8 sunscreen. Our data show that these two sunscreens provide levels of immune protection which exceed the levels predicted by their labeled SPFs in immunoprotection tests conducted in mice exposed to a relevant MISD of UV radiation from a source emitting a UV power spectrum similar to that of sunlight. PMID- 9225459 TI - In vivo photo-detection of chemically induced premalignant lesions and squamous cell carcinoma of the rat palatal mucosa. AB - Photo-detection using in vivo fluorescence was studied for different stages of chemically induced premalignant lesions and squamous cell carcinoma (SCC) of the Wistar rat palatal mucosa. It was found that the epithelial dysplasia (numerically expressed in the epithelial atypia index (EAI) of the rat palate, induced by repeated application of the carcinogen 4-nitroquinoline 1-oxide (4NQO), showed an increase approximately proportional to the duration of the application period. Photo-detection of the lesions using Photofrin-induced fluorescence was studied with dual-wavelength excitation and the subtraction of images, in an attempt to reduce the autofluorescence. The Photofrin dose was 2.5 mg kg-1. This was based on a dose-response study for normal tissue damage by photodynamic therapy (PDT) in this animal model, because the underlying rationale was to study photo-detection as a method of locating additional (early) malignancies in patients treated by PDT. Fluorescence intensities 24 and 48 h after injection of Photofrin were shown to increase with the duration of 4NQO application and with increasing EAI. For an EAI greater than 15, there was a statistically significant difference (p < 0.01) between the fluorescence signals obtained with and without the injection of Photofrin. Fluorescence signals of these lesions without the use of Photofrin (autofluorescence) also showed an increase with increasing stages of epithelial dysplasia of the rat palate. However, the fluorescence signals obtained with Photofrin were always higher than those of the autofluorescence. From this study, we conclude that photo-detection with Photofrin has potential in distinguishing chemically induced premalignant lesions and squamous cell carcinomas from the normal rat palatal mucosa. Photofrin (2.5 mg per kg of body weight) certainly adds to the sensitivity of photo-detection, but autofluorescence alone also has promising features for detecting premalignant and malignant lesions of the oral mucosa. PMID- 9225460 TI - Photodynamic efficacy of naturally occurring porphyrins in endothelial cells in vitro and microvasculature in vivo. AB - Photodynamic therapy (PDT) has been described in terms of cellular and vascular effects. The precise mechanisms of cellular and vascular damage are still unknown. In this study, the photodynamic inactivation of endothelial cells in vitro and damage to the microvasculature in vivo by naturally occurring porphyrins (uroporphyrin III (UP), coproporphyrin III (CP) and protoporphyrin IX (PP)) were investigated. The chick chorioallantoic membrane model (CAM model) was used, which is convenient for the study of damage to the microcirculation induced by PDT. The hydrophilic porphyrins UP and CP exhibited low cytotoxicity towards endothelial cells. Only small amounts of UP and CP were taken up, resulting in weak inactivation after irradiation. In contrast, the more lipophilic PP showed a marked cytotoxicity. Considerable amounts of PP were accumulated in the cells, leading to pronounced inactivation after light exposure. For the three porphyrins, damage to the microvasculature was observed. The damage caused by the hydrophilic porphyrins UP and CP was strongly dependent on the drug and light dose. For vascular injury, the efficacy was graded as UP < CP < PP. PMID- 9225461 TI - The plant ER: a dynamic organelle composed of a large number of discrete functional domains. AB - The endoplasmic reticulum (ER) of plants is comprised of a three-dimensional network of continuous tubules and sheets that underlies the plasma membrane, courses through the cytoplasm, and links up with the nuclear envelope. Aside from discussing the dynamic properties of this versatile and adaptable organelle, the review highlights the structure and the functional properties of 16 types of morphologically defined ER membrane domains. Owing to their labile or transient nature, several of these domains can only be visualized reliably through the use of ultrarapid freezing techniques. The ER domains discussed are: the lamin receptor domain; the nuclear pores; the nuclear envelope-ER gates, the microtubule nucleation domains; the protein and oil body-forming domains; the vacuole-forming ER; the actin-binding, the plasma membrane-anchoring and the vacuole and mitochondrion-attachment domains; the lipid recycling ER cisternae and the plasmodesmata. Preliminary evidence suggests that this list will have to be expanded in the near future. Understanding the assembly, the functional roles, and the developmental regulation of these domains has implications both for understanding cell structure and function, and for exploiting plants for agricultural and biotechnological purposes. PMID- 9225462 TI - Tryptophan decarboxylase is encoded by two autonomously regulated genes in Camptotheca acuminata which are differentially expressed during development and stress. AB - Camptothecin (CPT) is a valuable anti-cancer monoterpene alkaloid produced by the Chinese tree Camptotheca acuminata. Tryptophan decarboxylase (TDC) supplies tryptamine for the indole moiety of CPT and its derivatives, and is considered a key step in monoterpene indole alkaloid biosynthesis as it links primary and secondary metabolism. This report describes the isolation and characterization of tdc1 and tdc2, two autonomously regulated TDC genes from Camptotheca. When expressed in Escherichia coli, the products of each gene could decarboxylate tryptophan, but were inactive against tyrosine, phenylalanine and 3,4 dihydroxyphenylalanine (dopa), tdc1 was developmentally regulated, having its highest expression level in the apex, young stem and bark, tissues which also contain the highest levels of CPT. Expression of tdc1 also increased during seedling development and was correlated with alkaloid accumulation during germination. tdc2 expression was induced in Camptotheca leaf discs and cell suspension cultures treated with fungal elicitor or methyl jasmonate, treatments which did not affect tdc1 expression. Unlike tdc1, tdc2 expression was not detected in any unstressed Camptotheca tissues nor in developing seedlings. These data suggest that tdc1 may be part of a developmentally regulated chemical defense system in Camptotheca, while tdc2 serves as part of a defense system induced during pathogen challenge. PMID- 9225463 TI - Efficient gene targeting in the moss Physcomitrella patens. AB - The moss Physcomitrella patens is used as a genetic model system to study plant development, taking advantage of the fact that the haploid gametophyte dominates in its life cycle. Transformation experiments designed to target three single copy genomic loci were performed to determine the efficiency of gene targeting in this plant. Mean transformation rates were 10-fold higher with the targeting vectors and molecular evidence for the integration of exogenous DNA into each targeted locus by homologous recombination is provided. The efficiency of gene targeting determined in these experiments is above 90%, which is in the range of that observed in yeast and several orders of magnitude higher than previous reports of gene targeting in plants. Thus, gene knock-out and allele replacement approaches are directly accessible to study plant development in the moss Physcomitrella patens. Moreover, efficient gene targeting has so far only been observed in lower eukaryotes such as protozoa, yeasts and filamentous fungi, and, as shown here the first example from the plant kingdom is a haplobiontic moss. This suggests a possible correlation between efficient gene targeting and haplophase in eukaryotes. PMID- 9225464 TI - Characterization of a DNA-binding protein that interacts with 5' flanking regions of two fruit-ripening genes. AB - The E4/E8 binding protein (E4/E8BP) interacts with sequences in the 5' flanking regions of two genes, E4 and E8, that are coordinately regulated by ethylene during tomato fruit ripening. The DNA-binding activity of this protein increases during fruit ripening, and it may play a role in regulation of these genes. To begin to understand the function of this protein, a cDNA has been isolated that encodes a protein, E4/E8BP-1, with DNA-binding specificity similar to that of E4/E8BP. This DNA-binding protein is closely related to a DNA binding protein from tobacco, 3AF1, that interacts with the promoter of the pea rbcS-3A gene. A repeated domain was identified within the predicted 3AF1 amino acid sequence, which includes a series of histidines and cysteines, suggestive of zinc binding, and this repeat is conserved in E4/E8BP-1. Interaction of both E4/E8BP-1 and nuclear extracts from ripening fruit with the E8 recognition sequence is sensitive to 1,10-phenanthroline, indicating that a metal is required for binding of both the native and recombinant proteins. The mRNA for E4/E8BP-1 is moderately abundant in fruit, and increases slightly during fruit ripening, consistent with a role in fruit ripening. A truncated version of E4/E8BP-1 was able to transactivate the E4 promoter in transient assay, demonstrating that this DNA binding protein can interact with the E4 promoter in vivo to enhance gene transcription. PMID- 9225466 TI - A combinatorial role for exon, intron and splice site sequences in splicing in maize. AB - Plant introns are typically AU-rich or U-rich, and this feature has been shown to be important for splicing. In maize, however, about 20% of the introns exceed 50% GC, and most of them are efficiently spliced. A series of constructs has been designed to analyze the cis requirements for splicing of the GC-rich Bz2 maize intron and two other GC-rich intron derivatives. By manipulating exon, intron and splice site sequences it is shown that exons can play an important role in intron definition: changes in exon sequences can increase splicing efficiency of a GC rich intron from 17% to 86%. The relative difference, or base compositional contrast, in GC and U content between exon and intron sequences in the vicinity of splice sites, rather than the absolute base-content of the intron or exons, correlates with splicing efficiency. It is also shown that GC-rich intron constructs that are poorly spliced can be partially rescued by an improved 3' splice site. PMID- 9225465 TI - Isolation of a novel class of bZIP transcription factors that interact with ABA responsive and embryo-specification elements in the Dc3 promoter using a modified yeast one-hybrid system. AB - Dc3 is a carrot lea class gene that is abundantly expressed during somatic and zygotic embryogenesis. Its expression is normally embryo-specific and also can be induced by abscisic acid. The regulatory elements mediating the embryo-specific expression of Dc3 reside within the proximal promoter region (-117 to +26), which is also essential for ABA-induced expression. In this study, an optimized version of the yeast one-hybrid system has been used to clone factors that bind to the promoter region of the Dc3 gene. Twenty-five million yeast transformants were screened in a single experiment, and nine independent cDNA clones were isolated from a sunflower library that encode proteins that specifically bind to functional cis-regulatory elements in the Dc3 promoter. Analysis of these clones showed that they are derived from three different mRNA species that encode two basic leucine zipper proteins. The basic regions of these proteins, named DPBF-1 and 2 (Dc3 Promoter-Binding Factor-1 and 2), respectively, are nearly identical to each other and are similar to the plant G-box binding factor GBF-4. Outside the basic region, however, DPBF-1 and 2 diverge significantly from each other and from other known factors. Both factors have transcriptional activity in yeast, and bind to DNA as dimers. Unlike other plant bZIP factors, DPBF-1 and 2 recognize sequences containing the ACACNNG core. Cloning of these factors demonstrates the power of the one-hybrid approach when optimally applied. PMID- 9225468 TI - Regulatory elements in vivo in the promoter of the abscisic acid responsive gene rab17 from maize. AB - The rab17 gene from maize is transcribed in late embryonic development and is responsive to abscisic acid and water stress in embryo and vegetative tissues. In vivo footprinting and transient transformation of rab17 were performed in embryos and vegetative tissues to characterize the cis-elements involved in regulation of the gene. By in vivo footprinting, protein binding was observed to nine elements in the promoter, which correspond to five putative ABREs (abscisic acid responsive elements) and four other sequences. The footprints indicated that distinct proteins interact with these elements in the two developmental stages. In transient transformation, six of the elements were important for high level expression of the rab17 promoter in embryos, whereas only three elements were important in leaves. The cis-acting sequences can be divided in embryo-specific, ABA-specific and leaf-specific elements on the basis of protein binding and the ability to confer expression of rab17. We found one positive, new element, called GRA, with the sequence CACTGGCCGCCC. This element was important for transcription in leaves but not in embryos. Two other non-ABRE elements that stimulated transcription from the rab17 promoter resemble previously described abscisic acid and drought-inducible elements. There were differences in protein binding and function of the five ABREs in the rab17 promoter. The possible reasons for these differences are discussed. The in vivo data obtained suggest that an embryo specific pathway regulates transcription of the rab genes during development, whereas another pathway is responsible for induction in response to ABA and drought in vegetative tissues. PMID- 9225467 TI - A homolog of the Arabidopsis thaliana ERS gene is actively regulated in Rumex palustris upon flooding. AB - A cDNA homologous to the ethylene-response sensors (ERS/ETR1) from Arabidopsis thaliana was isolated from a Rumex palustris cDNA library. This cDNA, RP-ERS1, was 2421 bp long and shared 66% nucleotide homology with ETR1 and ERS in their coding regions. The transcript level of RP-ERS1 was actively regulated during the leafelongation response of R. palustris upon flooding. RP-ERS1 transcript levels increased after submergence, and also after exposure to high concentrations of ethylene and carbon dioxide and low concentrations of oxygen. These results suggest that R. palustris plants may respond to flooding stress by increasing the number of their ethylene receptors. PMID- 9225469 TI - A novel nucleic acid helicase gene identified by promoter trapping in Arabidopsis. AB - A gene encoding a predicted nucleic acid helicase was identified in Arabidopsis thaliana by activation of a promoter trap. An in vivo transcriptional fusion between the helicase gene and a promoterless uidA (gusA) gene was generated, which was expressed specifically in the tapetum and vascular tissues. The tagged gene, designated HVT1 (Helicase in Vascular tissue and Tapetum), encodes a native transcript of approximately 4.4 kb, which is of very low abundance. The predicted HVT1 protein, of 1291 amino acid residues, is homologous to the Drosophila MALELESS, human RNA helicase A and bovine nuclear DNA helicase proteins, and represents the first identified member of a new subgroup within the mle group of the DEAH family. Low stringency genomic Southern blot analysis indicates that at least two structurally related genes exist in Arabidopsis. We suggest that the HVT1 protein may play a role in nucleic acid unwinding in restricted cell types during both vegetative and reproductive phases of the Arabidopsis life cycle. PMID- 9225470 TI - Phosphorylation of nuclear proteins directs binding to salicylic acid-responsive elements. AB - The cis-located DNA sequence as-1 (Activation Sequence-1) from CaMV 35S promoter has been previously identified as an element that can confer inducibility by salicylic acid (SA) with immediate early kinetics. This sequence specifically binds to ASF-1 (Activation Sequence Factor-1), previously characterized in tobacco nuclear extracts. To assess whether modulation of ASF-1 binding activity can explain the activation of the as-1 sequence observed in vivo, we performed electrophoretic mobility shift assays using nuclear extracts from SA-treated and water-treated tobacco plants. Our results indicate that treatment of plants with SA increases ASF-1 binding to as-1 and to ocs, an as-1-like element from the Agrobacterium octopine synthase gene. In contrast, SA treatment has no effect on the binding of GT-1 factor to its target light-inducible box II element. Furthermore, treatment of nuclear extracts from SA-treated plants with alkaline phosphatase decreases ASF-1 binding to the as-1 element. This can be reversed by pretreatment with 10 mM NaF. Accordingly, pretreatment of nuclear extracts from control water-treated plants with ATP produces an increase in ASF-1 binding activity similar to that observed with SA. This effect of ATP is reversed by treatment with alkaline phosphatase and prevented by quercetin, a casein kinase II inhibitor. These results support the hypothesis that a nuclear protein kinase is involved in the immediate early events of transcriptional activation triggered by SA. PMID- 9225471 TI - An Arabidopsis thaliana cDNA complementing a hamster apoptosis suppressor mutant. AB - Programmed cell death or apoptosis is a process in which unwanted cells are eliminated during growth and development. In mammals, several genes have been identified whose products are necessary to prevent entry into the apoptotic process. We have isolated a clone from an Arabidopsis thaliana cDNA library whose predicted translation product shows highly significant similarity to the mammalian defender against apoptotic death 1 (DAD1) protein. Transformation of the mutant hamster tsBN7 cells, which undergo apoptosis at restrictive temperature, demonstrates that the plant protein is as efficient as human DAD1 in rescuing these hamster cells from apoptosis. In contrast to mammals, Southern hybridisation and genomic data indicate that there are probably two genes in Arabidopsis thaliana. Northern blot analysis shows that AtDAD transcripts are present in all tissues examined, although the abundance of the transcripts is reduced in siliques during the maturation and desiccation phase of the seed. This is the first experimental proof that a homologue of an animal gene involved in apoptosis exists in plants and the first demonstration of complementation of a vertebrate mutant by a plant cDNA. Our results suggest that this process of suppression of apoptosis has been conserved in animals and plants. PMID- 9225472 TI - A promoter trap for Chlamydomonas reinhardtii: development of a gene cloning method using 5' RACE-based probes. AB - A promoterless radial spoke protein RSP3 gene has been used to identify promoter regions in the genome of Chlamydomonas reinhardtii. The acceptor strain pf-14 arg7 was transformed with a linearized vector containing the ARG 7.8 gene as a selection marker and a promoterless RSP3 gene. The frequency at which the motility was restored in transformants varied from 2-3%. Several of these were motile only in ammonium-free medium, indicating that the procedure could be used to select inducible promoters. Transformation of nitrogen-starved cells produced about twice as many transformants which were only motile in ammonium-free medium. Since one of the tagging vectors contained an RSP3 gene with a hybridization flag in its 3' untranslated region, it was possible to estimate the size of the new RSP3 transcripts in transformants. The results suggested that in most cases a hybrid RNA was generated consisting of the tagged gene transcript and reporter gene RNA. By 5' RACE, these parts of the new transcripts were amplified and it was shown that the generated DNA fragments could be used to clone a tagged gene. One such example, gene 2BC9, is predicted to code for a mitochondrial matrix protein. The tagging procedure will be optimized for cloning genes induced by nitrogen starvation, the cue for gametogenesis. PMID- 9225473 TI - Retrofitting YACs for direct DNA transfer into plant cells. AB - The utility of plant YAC libraries prepared in conventional YAC vectors would be dramatically increased if these YACs could be used directly for plant transformation. A pair of vectors that allow clones from YAC libraries to be modified (retrofitted) for plant transformation by direct DNA transfer methods, such as particle bombardment or electroporation, has been developed. Modification of the YAC is achieved in two sequential yeast transformation steps by taking advantage of the homologous recombination system in yeast. Using this approach, two plant-selectable marker genes and DNA sequence elements required for copy number amplification in yeast can be introduced into YACs present in yeast strain AB1380. The utility of these vectors is demonstrated by retrofitting YACs that contain inserts ranging in size from 80 to 700 kb. The 6- to 12-fold increase in copy number of these modified YACs facilitates the isolation of YAC DNA for direct DNA transformation methods. Retrofitted YACs were used for particle bombardment to examine the efficiency with which their large DNA inserts are transferred into plant cells. The availability of these retrofitting vectors should facilitate the transfer of YAC DNA inserts into plant cells and thus help bridge the gap between existing mapping techniques and plant transformation procedures. PMID- 9225474 TI - Monoclonal antibodies to barley aleurain and homologs from other plants. AB - Barley aleurain is contained within a specific type of vacuole characterized by acidic pH and the presence of other hydrolytic enzymes. The aleurain-containing vacuole is distinct from protein storage vacuoles, and anti-aleurain antibodies serve as markers for this organelle in barley cells. Aleurain is a unique type of cysteine protease, and other plant species have genes for homologs whose sequences are highly conserved, but little is known about these homologs at the protein level. Seven monoclonal antibodies to barley aleurain were isolated, which bind to and define aleurain homologues in Arabidopsis, Petunia, and tobacco cell extracts. Interestingly, in addition to 29-32 kDa aleurain homologs, Petunia extracts contain a protein of approximately 50 kDa and tobacco extracts a protein of approximately 40 kDa that are recognized by multiple different monoclonal antibodies, indicating an unexpected diversity to the aleurain protein family. Among the group of antibodies are some that efficiently immunoprecipitate metabolically labeled aleurain from barley cell extracts, and some that efficiently label aleurain in immunofluorescence assays using root tip cells. These antibodies should be useful for plant cell biologists who study vacuole biogenesis and function and sorting of proteins to specific vacuolar compartments, in barley as well as other plant species. PMID- 9225476 TI - Families of stationary patterns producing illusory movement: insights into the visual system. AB - A computational explanation of the illusory movement experienced upon extended viewing of Enigma, a static figure painted by Leviant, is presented. The explanation relies on a model for the interpretation of three-dimensional motion information contained in retinal motion measurements. This model shows that the Enigma figure is a special case of a larger class of figures exhibiting the same illusory movement and these figures are introduced here. Our explanation suggests that eye movements and/or accommodation changes cause weak retinal motion signals, which are interpreted by higher-level processes in a way that gives rise to these illusions, and proposes a number of new experiments to unravel the functional structure of the motion pathway. PMID- 9225478 TI - Variation in growth form and precocity at birth in eutherian mammals. AB - Using the flexible Chapman-Richards model for describing the growth curves from birth to adulthood of 69 species of eutherian mammals, we demonstrate that growth form differs among eutherian mammals. Thereby the commonly used Gompertz model can no longer be considered as the general model for describing mammalian growth. Precocial mammals have their peak growth rate earlier in the growth process than altricial mammals. However, the position on the altricial-precocial continuum accounts for most growth-form differences only between mammalian lineages. Within mammalian genera differences in growth form are not related to precocity at birth. This indicates that growth form may have been associated with precocity at birth early in mammalian evolution, when broad patterns of body development radiated. We discuss four non-exclusive interpretations to account for the role of precocity at birth on the observed variation in growth form among mammals. Precocial and altricial mammals could differ according to (i) the distribution of energy output by the mother, (ii) the ability of the young to assimilate the milk yield, (iii) the allocation of energy by the young between competing functions and (iv) the position of birth between conception and attainment of physical maturity. PMID- 9225477 TI - Transitions in individuality. AB - The evolution of multicellular organisms is the premier example of the integration of lower levels into a single, higher-level individual. Explaining the evolutionary transition from single cells to multicellular organisms is a major challenge for evolutionary theory. We provide an explicit two locus genetic framework for understanding this transition in terms of the increase of cooperation among cells and the regulation of conflict within the emerging organism. Heritability of fitness and individuality at the new level emerge as a result of the evolution of organismal functions that restrict the opportunity for conflict within and ensure cooperation among cells. Conflict leads, through the evolution of adaptations that reduce it, to greater individuality and harmony for the organism. PMID- 9225475 TI - Dynamics of two feline retroviruses (FIV and FeLV) within one population of cats. AB - We present a deterministic model of the dynamics of two microparasites simultaneously infecting a single host population. Both microparasites are feline retroviruses, namely Feline Immunodeficiency Virus (FIV) and Feline Leukaemia Virus (FeLV). The host is the domestic cat Felis catus. The model has been tested with data generated by a long-term study of several natural cat populations. Stability analysis and simulations show that, once introduced in a population, FIV spreads and is maintained, while FeLV can either disappear or persist. Moreover, introduction of both viruses into the population induces an equilibrium state for individuals of each different pathological class. The viruses never induce the extinction of the population. Furthermore, whatever the outcome for the host population (persistence of FIV only, or of both viruses), the global population size at the equilibrium state is only slightly lower than it would have been in the absence of the infections (i.e. at the carrying capacity), indicating a low impact of the viruses on the population. Finally, the impact of the diseases examined simultaneously is higher than the sum of the impact of the two diseases examined separately. This seems to be due to a higher mortality rate when both viruses infect a single individual. PMID- 9225479 TI - Contribution of NMDA and non-NMDA glutamate receptors to locomotor pattern generation in the neonatal rat spinal cord. AB - The motor programme executed by the spinal cord to generate locomotion involves glutamate-mediated excitatory synaptic transmission. Using the neonatal rat spinal cord as an in vitro model in which the locomotor pattern was evoked by 5 hydroxytryptamine (5-HT), we investigated the role of N-methyl-D-aspartate (NMDA) and non-NMDA glutamate receptors in the generation of locomotor patterns recorded electrophysiologically from pairs of ventral roots. In a control solution, 5-HT (2.5-30 microM) elicited persistent alternating activity in left and right lumbar ventral roots. Increasing 5-HT concentration within this range resulted in increased cycle frequency (on average from 8 to 20 cycles min-1). In the presence of NMDA receptor antagonism, persistent alternating activity was still observed as long as 5-HT doses were increased (range 20-40 microM), even if locomotor pattern frequency was lower than in the control solution. In the presence of non NMDA receptor antagonism, stable locomotor activity (with lower cycle frequency) was also elicited by 5-HT, albeit with doses larger than in the control solution (15-40 microM). When NMDA and non-NMDA receptors were simultaneously blocked, 5 HT (5-120 microM) always failed to elicit locomotor activity. These data show that the operation of one glutamate receptor class was sufficient to express locomotor activity. As locomotor activity developed at a lower frequency than in the control solution after pharmacological block of either NMDA or non-NMDA receptors, it is suggested that both receptor classes were involved in locomotor pattern generation. PMID- 9225480 TI - Phylogenetic survey of soluble saxitoxin-binding activity in pursuit of the function and molecular evolution of saxiphilin, a relative of transferrin. AB - Saxiphilin is a soluble protein of unknown function which binds the neurotoxin, saxitoxin (STX), with high affinity. Molecular characterization of saxiphilin from the North American bullfrog, Rana catesbeiana, has previously shown that it is a member of the transferrin family. In this study we surveyed various animal species to investigate the phylogenetic distribution of saxiphilin, as detected by the presence of soluble [3H]STX binding activity in plasma, haemolymph or tissue extracts. We found that saxiphilin activity is readily detectable in a wide variety of arthropods, fish, amphibians, and reptiles. The pharmacological characteristics of [3H]STX binding activity in phylogenetically diverse species indicates that a protein homologous to bullfrog saxiphilin is likely to be constitutively expressed in many ectothermic animals. The results suggest that the saxiphilin gene is evolutionarily as old as an ancestral gene encoding bilobed transferrin, an Fe(2+)-binding and transport protein which has been identified in several arthropods and all the vertebrates which have been studied. PMID- 9225481 TI - DNA phylogeny of the marsupial wolf resolved. AB - The phylogenetic position of the recently extinct marsupial 'wolf', or thylacine (Thylacinus cynocephalus), has been a source of contention in mammalian systematics for nearly a century. Thylacines were endemic to Australasia, but possessed striking anatomical similarities to Oligo-Miocene borhyaenid marsupials of South America. At issue has been whether these features are indicative of common ancestry or convergent adaptation to carnivory. Recent morphological studies have supported both conclusions. Although current marsupial classifications group thylacines with Australian dasyuromorphians, this putative clade is characterized by mostly primitive morphological features. Attempts to determine thylacine affinities with ancient protein and DNA analyses have supported, but not resolved, a dasyuromorphian placement. We report 1546 bp of mitochondrial DNA sequence (from cytochrome b and 12S rRNA genes) and 841 bp of nuclear protamine gene sequence from the thylacine and representatives of all or most other marsupial orders. Phylogenetic analysis of these sequences shows unambiguously that thylacines are members of Dasyuromorphia, and suggests a late Oligocene or very early Miocene divergence of familial lineages. PMID- 9225482 TI - Mixed-genotype infections of malaria parasites: within-host dynamics and transmission success of competing clones. AB - Mixed-genotype infections of microparasites are common, but almost nothing is known about how competitive interactions within hosts affect the subsequent transmission success of individual genotypes. We investigated changes in the composition of mixed-genotype infections of the rodent malaria Plasmodium chabaudi clones CR and ER by monoclonal antibody analysis of the asexual infection in mice, and by PCR amplification of clone-specific alleles in oocysts sampled from mosquitoes which had fed on these mice. Mixed-clone infections were initiated with a 9:1 ratio of the two clones, with ER as the minority in the first experiment and CR as the minority in the second experiment. When beginning as the majority, clones achieved parasite densities in mice comparable to those achieved in control (single-clone) infections. When they began as the minority, clones were suppressed to less than 10% of control parasitaemias during the early part of the infections. However, in mosquitoes, the frequency of the initially rare clone was substantially greater than it was in mice at the start of the infection or four days prior to the feed. In both experiments, the minority clone in the inocula produced as many, or more, oocysts than it did as a single-clone infection. These experiments show that asexual dominance during most of the infection is poorly correlated to transmission probability, and therefore that the assumption that within-host population size correlates to transmission probability may not be warranted. They also raise the fundamental question of why transmission rates of individual genotypes are often higher from mixed than single-clone infections. PMID- 9225484 TI - Histological and ultrastructural study of the effects of cholinergic and adrenergic agonists on salivary secretion from the lingual epithelium and the lingual gland of the Tokyo Daruma pond frog. AB - Comparative observations of the effects of cholinergic and adrenergic agonists (such as pilocarpine, phenylephrine and isoproterenol) on the secretion of salivary fluid and of secretory granules from the lingual epithelium and the lingual gland in the Tokyo Daruma pond frog, Rana porosa porosa, were made by light and transmission electron microscopy. The effect of pilocarpine on the loss of cytoplasm in both the lingual epithelium and the lingual gland was the strongest, and that of isoproterenol was the weakest. Transmission electron microscopy revealed that electron-dense granules, located in cells in both the lingual epithelium and the lingual gland, were discharged by exocytosis after stimulation by phenylephrine and isoproterenol. Immediately before secretion of these granules, they became somewhat larger and round or distorted in shape. However, after administration of pilocarpine, no obvious discharge of electron dense granules was apparent and, instead, granules in some cells began deteriorating within the cells and those in other cells developed electron-dense and electron-lucent areas. A dot-like pattern was recognized in the electron dense areas of these granules. These phenomena were assumed to be secondary effects accompanying the secretion of salivary fluid from cells of the lingual epithelium and the lingual gland. By contrast, mucous granules in the lingual gland were secreted by a holocrine process. PMID- 9225483 TI - Microvessels of the chick chorioallantoic membrane uniformly restrict albumin extravasation during angiogenesis and endothelial cytodifferentiation. AB - The present study served to evaluate extravasation of albumin across the microvascular endothelium of the chick chorioallantoic membrane (CAM) during the establishment of endothelial barrier functions (days 4.5 to 6.0), during a rapid phase of normigenesis (day 10), and after initiation of endothelial cytodifferentiation (day 14). CAM preparations in shell-less cultures were evaluated by intravital fluorescence confocal microscopy and videodensitometry after microinjections of chicken serum albumin (CSA) conjugated to Fluorescein isothiocyanate (FITC) (1% in Avian Ringers). At each observed temporal stage, FITC-CSA extravasation from the CAM pre-capillaries, capillaries, and post capillaries was uniformly negligible. Since the endothelial glycocalyx imparts a net negative luminal charge, impedance of CSA transport by mutual charge repulsion was also evaluated. Accordingly, CAM microvessels were pre-infused with protamine sulfate (100 microliters/ml) to neutralize the negative charge. However, interstitial accumulation of FITC-CSA remained negligible. Thus, the presence of tight junctional clefts and paucity of plasmalemmal vesicles during CAM normigenesis probably serve as the principal morphologic correlates to the observed albumin restriction. Further, the normigenic CAM endothelium presents a more restrictive barrier to macromolecules than that of most angiogenic endothelia. PMID- 9225485 TI - Innervation of taste buds in the canine larynx as revealed by immunohistochemistry for the various neurochemical markers. AB - The distribution and innervation of the canine laryngeal taste buds were observed using immunohistochemistry with antibodies against protein gene product 9.5 (PGP 9.5) and neurofilament protein (NFP). We also observed the immunohistochemical distribution of serotonin, tyrosine hydroxylase (TH) and various neuropeptides including calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal peptide (VIP), galanin, methionine enkephalin (ENK) and neuropeptide Y (NPY). The taste buds in the canine larynx were densely distributed in the mucosa at the basal portion of the epiglottis and cuneiform process of the arytenoid cartilage. The taste cells were immunoreactive for PGP 9.5 and serotonin. The nerve fibers with immunoreactivity for PGP 9.5 in the taste buds were observed in the perigemmal region and intra- and subgemmal plexuses, and these were classified into two types based on their diameter. The thick nerve fibers corresponded to the fibers immunoreactive for NFP, while the thin nerve fibers corresponded to the fibers immunoreactive for TH and various neuropeptides. Numerous nerve fibers immunoreactive for SP and CGRP were observed in the perigemmal region, and intra- and subgemmal plexuses. A few galanin- and ENK immunoreactive nerve fibers were also observed in the taste buds, whereas NPY immunoreactive nerve fibers were noted beneath them. All peptide-containing fibers except for VIP-immunoreactive nerves were situated in the subgemmal regions. In conclusion, the multiple innervation to the laryngeal taste buds were documented. Thick nerve fibers are likely to be irritant receptors, while thin varicose nerve fibers seem to regulate taste buds themselves. The laryngeal taste buds may be among the important structures which are sensitive to exogeneous chemical and/or mechanical stimuli, for the protection of the airway and the regulation of the respiratory function. PMID- 9225486 TI - Distribution of sulfakinin-like peptides in the central and sympathetic nervous system of the American cockroach, Periplaneta americana (L.) and the field cricket, Teleogryllus commodus (Walker). AB - We describe the distribution of sulfakinin-like neuropeptides in the central and sympathetic nervous system of the American cockroach Periplaneta americana (L.) (Blattodea) and the field cricket Teleogryllus commodus (Walker) (Othoptera), using an antisulfakinin primary antibody and confocal laser scanning microscopy. We conclude that, in the cockroach, sulfakinin-like material is produced in ten pairs of anterior cells in the pars intercerebralis, as well as two pairs of medial and one major pair of lateral posterior brain cells. This contrasts with findings in other insects, including the cricket, where only the posterior cell groups express sulfakinin-immunoreactive material. Extensive arborization of dendrites containing sulfakinin-like peptides occurs within the neuropile of both species, suggesting a neurotransmitter/neuromodulator function. In the cockroach, there is clear evidence of direct distribution of sulfakinin-like peptides along axons to the foregut tissue, and a plexus of retrocerebral nerves is likely to serve as a neurohaemal release site. Neurohaemal release into the dorsal aorta is also postulated. Sulfakinin-immunoreactive axons do not innervate the hindgut in either cockroaches or crickets. Sulfakinin may function as a gut myotropin in the Blattodea, in addition to functioning as a neurotransmitter within the central nervous system. This latter function appears to be general across insect orders, while the neurohaemal distribution and myotropic activity are restricted to the Blattodea. PMID- 9225487 TI - Islets and diffuse endocrine component in the pancreas of three red frogs species: relationships between endocrine and exocrine tissue. AB - The endocrine pancreas of three red frogs was studied immunohistochemically. It consisted of islets and diffuse endocrine cells. The islets showed a mammalian like arrangement with a central core of B cells and a peripheral mantle of A/PP cells. A few D and VIP cells were also present. Several regulatory peptides were co-localized in the same endocrine cells by consecutive sections and double labeling studies. The A/PP cells were formed by subpopulations of cells showing various types of immunoreactivity and varying degrees of immunolabeling. Generally, glucagon/pancreatic polypeptide, glucagon/pancreatic polypeptide/peptide tyrosine tyrosine and glucagon/pancreatic polypeptide/neuropeptide tyrosine immunoreactivities were present in the islets and in the endocrine cells scattered throughout the exocrine parenchyma (the diffuse component). Some specimens, mainly belonging to Rana dalmatina, showed evident periinsular halos around the islets. The diffuse component was abundant, and mainly contained A/PP cells. It formed a net across the exocrine parenchyma; its interrelationship with the latter might occur by a paracrine mechanism. PMID- 9225488 TI - Significance of the localization of large granules in the thyrogonadotrophs of the musk shrew (Suncus murinus). AB - In order to examine the significance of small round and large granules in the thyrogonadotrophs of the musk shrew, we investigated the differences of numbers, distribution patterns and/or immunoreactivities of the large granules in the thyrogonadotrophs among the newborn, infant and adult musk shrews. As a result, we observed that the thyrogonadotrophs exist in newborn musk shrew, but the large granules were few in number and immunoreactivity on the granules was weak. The ratio of the number of large granule-containing cells to all LH beta positive cells was very low (6.2%) in newborn musk shrew, but high (98.2%) in adult male animals. We assume that thyrogonadotrophs of newborn shrews may not yet synthesize active beta-subunits, that the synthesis of beta-subunits is gradually increased with aging, and excessive products of beta-subunits may be stored in the large granules. It is known that female musk shrews are induced ovulators; therefore, we investigated morphological changes in the large and small granules in the thyrogonadotrophs at the LH surge which occurs after mating. We observed that the small granules were markedly decreased in number, or had almost vanished, 2 h after mating, but that the large granules still remained. Consequently, we assume that the large granules may be storage sites of excessive products of beta-subunits in older animals. On the other hand, small granules (release type granules) may be exhausted by release of a large quantity of gonadotrophin 2 h after mating, and this acute exhaustion of small granules may induce a sharp depression of the LH surge. PMID- 9225490 TI - Ambulatory blood pressure measurement: what is the use? PMID- 9225489 TI - Financial equivalent, industry sponsored research and conflict of interest. AB - Conflicts of interest for the clinician(physician)-researcher are not limited only to direct and clear financial support by manufacturers of the pharmaceutical and medical device industry, but rather include delicate indirect monetary and research support. Today professionals face an inevitable choice between two opposing moral orders, one based in the primacy of ethical obligations to the sick, the other in the primacy of self-interest and the marketplace. Some medical ethicists urge, reshape ethical codes to conform to the ethos of the marketplace, which legitimates self-interest over beneficence and makes vices out of most of traditional virtues. Second opinion represents the ethicists who recommend a firm stand in belief that being a physician imposes certain specific obligations. Medicine is at heart a moral enterprise and those who practice it are de facto members of a moral community. The market introduces an alien-till this time unknown-set of economic values into an institution (medicine) whose inherent ends are altruistic, but in countries under health care reform it brings a complex of special ethical issues in connection with deficient legislation and not firm ethical rules adopted. (Ref. 17.) PMID- 9225492 TI - Respiratory sleep disorders and high blood pressure. PMID- 9225491 TI - Carbonic anhydrase and the heart. PMID- 9225493 TI - Assessment of left ventricular systolic function in low-echogenic patients by intravenous Infoson injection during dopamine echocardiography. An open, phase III trial. AB - This trial evaluated whether the intravenous injection of an ultrasound contrast agent (Infoson) facilitates the assessment of systolic function in 40 low echogenic patients undergoing low-dose (4 mcg/kg/min) dopamine echocardiography. Interobserver difference in calculated ejection fraction at entry was > 10%. Echocardiographic monitoring was performed in the apical 4-chamber view at four intervals: baseline, no contrast; first Infoson injection; dopamine infusion, no contrast; dopamine infusion+second Infoson injection. The left ventricle was divided into 5 segments and analysis was performed by two blinded observers. Wall motion abnormalities, ejection fraction and the confidence in detecting the endocardial border, were assessed. Infoson provided adequate left ventricular opacification in 90% of the injections, with a significant improvement in endocardial border detection. The interobserver variability of ejection fraction measurements was significantly reduced. The probability of attaining concordance between the investigators on wall motion assessment improved significantly. These results suggest potential applications in stress echocardiography. PMID- 9225495 TI - Transdisciplinarity in the study of undernutrition-infection interactions. AB - Interactions between undernutrition, infection, and growth and development are complex, and are reviewed in this article, giving particular emphasis on the importance of diarrheal infection in this process. The effects of diet, nutrition and infection on the nutritional status of a child can vary according to the disease ecology, the age of the child, patterns of feeding and types of food consumed. There are two possible ways in which this relationship can begin; one in which poor nutritional status leads to impaired immunocompetence and reduced resistance to infection, and the other in which exposure to infectious disease can lead to appetite loss and anorexia, malabsorption, and elevated metabolism of energy and other nutrients. Once started, the interactions between these two major environmental stressors becomes increasingly complex, with the nature of the disease ecology influencing the balance of immunoparesis and adaptive immunity and its effect on subsequent disease experience. Furthermore, the disease ecology influences the type and extent of associated physiological phenomena including anorexia, fever, and malabsorption, all of which have an impact on nutritional status. Of disease categories, diarrhea has particularly potent effects in this relationship. The predicted impact of HIV infection among newborn infants is the earlier onset of the undernutrition-infection cycle, as low CD4+ T lymphocyte counts soon after birth are likely to predispose such infants to earlier opportunistic infection. PMID- 9225496 TI - Physiological consequences of everyday psychosocial stress. AB - A large body of data has been accumulated concerning physiological responses in people exposed to stressors in laboratories. Adrenaline and cortisol have become known as "stress hormones" because, in men, levels of both hormones consistently rise in response to stress in laboratory-based investigations. If chronically repeated, elevation of adrenaline and cortisol is likely to have long-term consequences for health, especially cardiovascular health, partly via the effects of the hormones on blood pressure and serum cholesterol levels. Research on people conducting their everyday lives is necessary to establish whether the same responses are shown on a day to day basis. Such research requires new methodologies and careful data collection. So far, it has been shown that adrenaline and blood pressure do seem to vary in expected ways. Other responses in everyday life, including those of cholesterol, cortisol and the immune system, are less well characterised. PMID- 9225497 TI - Understanding the nutrition of poor urban women: ethnographic and biological approaches. AB - To better understand how women respond to conditions of urban poverty in a developing country a sample of 85 women living in Cali, Colombia was studied. Anthropometric indicators of nutritional status were normal for the group. However, many women indicated that they did not always have sufficient money to purchase food, and described the strategies they used when financial resources were inadequate. These strategies included changes in meal composition, reductions in food portion size, and reductions in the number of meals eaten. Evidence of the use of these strategies was identified in 17.1% of all diet records (n = 509). The adequate nutritional status of this group of women suggests that their strategies were usually successful in maintaining adequate energy intake, but the frequent use of these strategies suggests that the women are potentially at risk for undernutrition. PMID- 9225498 TI - Transdisciplinary approach in women's health research: a study on urban middle class women. AB - The paper discusses the importance of adopting a transdisciplinary approach in women's health research and highlights the important issues relating to women's health. Specific health problems of women in developing countries have been outlined, and some findings of research on the relationship between working status and maternal health among the urban middle-class women of Calcutta are reported. They do not support the hypothesis that working women are under greater stress and hence suffer from ill health. The availability of hired domestic help and the support received from other family members may help mitigate the burden. PMID- 9225500 TI - Biocultural approaches to health and mortality in an Old Order Amish community. AB - The assumption that Amish cultural value of cooperation leads to greater longevity, prosperity and well-being among elders was examined using historical demographic and ethnographic analysis in a conservative Old Order Amish community. Migration, fertility, mortality and morbidity data since 1948 were used to identify population structure. The population pyramid in the Amish community (3% of the population over age 60, compared to 18% of the neighboring non-Amish rural area) is largely determined by natural fertility with low infant mortality. Household ethnography explored health beliefs, access to health care, caregiving patterns, and economic strategies. The perception that community cooperation and altruistic behavior were of benefit to Amish health is supported by mortality rates. Amish age standardized death rates were 19% below the US death rate in 1960. Migrating to form new communities and selectively utilizing acute (but not preventive or public) health care services, emerged as strong cultural patterns that facilitate reproductive success among the Amish. PMID- 9225499 TI - An universal model of health and work output: theoretical structure and testing strategy. AB - The model proposed in this paper presents a broad range of factors to predict individual human work output. The predictors include aerobic capacity, body size, motivation, work pattern, social environment and social network, caloric intake, drug and alcohol use, stress resistance and thermoregulation. Health is a major intervening variable, and its relationship to work output is a special concern of this research. We suggest that this model may be used as a template to explain human productivity in most societies. Its universality can be subjected to rigorous testing in a range of settings from tropical upland swidden horticulturalists to urban workers in a northern industrialized country. Observations are offered on some of those testing sites and on methodological issues implicit in research of this breadth. A major pilot study of urban Chinese workers has already demonstrated the predictive power of the model in one setting. PMID- 9225501 TI - Aging and gerontology: a paradigm of transdisciplinary research. AB - During the latter half of the twentieth century many societies experienced rapid increases in the number of persons aged 65 years and older. Such increases followed world wide reductions in infant and childhood mortality and the widespread control of infectious diseases through sanitation and medical advances. Today, we observe increasingly older average ages in many industrialized cosmopolitan societies, some approaching 35 years. With this increase, the need to understand the problems of elders and research on age related patterns of health change also have increased. This paper explores some of the advantages that a transdisciplinary approach might have to studies of aging and gerontology. Particular emphasis is placed on the chronic degenerative diseases, activities of daily living, and longitudinal patterns of individual change. PMID- 9225502 TI - Health promotion of adolescents. AB - China has been making progress in adolescence health care, carrying out directed investigations and academic exchanges, as well as training. Since 1949, both growth and development of Chinese children and adolescents have accelerated significantly. Menarche and the secondary sex characteristics of girls now appear earlier than before. The average age of menarche is 12.5 years (1991) and boys average first emission is 14.33 years (1991). In China, the commonly encountered adolescent health problems are menstruation hygiene, menstruation dysfunction, emission, masturbation, teenage pregnancy, acne, obesity, smoking, alcohol drinking, drug abuse, and suicide. Causes of death of adolescents in China has significantly changed, all deaths caused by infectious diseases have dropped significantly. Of all death causes today, accidental injury is the leading one. Sexually transmitted diseases and tuberculosis have shown a rebound recently. The rate of smoking among middle school students in Beijing increased from the 1980s to 1990s, with male students' smoking at significantly higher rates than female students. Adolescents is a transitional period from dependent childhood to independent adulthood. Good physical and mental health of children and adolescents makes for good health in adulthood, therefore adolescence is a very important period in one's life. We need to go a step further and develop more detailed data on adolescent health and provide more health care for adolescents. PMID- 9225504 TI - Maximal aerobic capacity and its relationship with physical growth in Chinese children. AB - Maximal aerobic capacity and its relationship to physical growth in 463 Chinese children and adolescents aged 10-19 is reported. Results show that VO2max, VO2max/height and VO2max/HRmax positively relate to measures of physical growth such as height, weight and lean body mass (LBM). Correlation of VO2max/weight with physical growth variables is high and positive in boys and low and negative in girls. The sample is divided to two groups on each variable: well-developed and relatively poorly-developed according to height, weight and LBM. SDS values of VO2max, VO2max/height and VO2max/HRmax are higher in the first group than in the second group. In contrast, SDS values of VO2max/weight and VO2max/LBM are greater in second group. The similarity of results between SDS values with correlation analysis suggest that weight and LBM are probably the decisive factors that influence VO2max. PMID- 9225503 TI - The status of physical growth in Chinese children. AB - This paper reports the physical growth status of about a half million Chinese children covering 28 nationalities in 1985. There are 4 items (height, weight, sitting-height, and chest circumference) that have been analyzed. Results are as follows: Han children display comparatively high physical growth levels. A growth difference emerges between Han Children who live in different social and geographical environments. The growth of urban children is better than that of rural children; the growth of children living in north China is better than that of children in south China. Differences between administrative regions are rather large. Beijing, Tianjin, Shanghai, Shandong, Liaoning and Heilongjiang display high-level growth; Guizhou, Qinghai, Sichuan, Yunnan, Fujian provinces and Guangxi autonomous region display low-level growth. The growth differences between minority nationalities are noticeable. Uighur, Kazak, Khalkhas, Hui, Mongol, and Korean, nationalities demonstrate relatively high growth level; Lahu, Wa, Yao, Tong, Miao, Puyi and She nationalities appear to maintain a low growth level. The probable reasons for the differences are discussed. PMID- 9225505 TI - The aging process--an analysis of the latent structure of body morphology (in males). AB - The morphological characteristics (20 anthropometric variables) of a total of 2,351 examinees (from the age of 18 to 90) were analyzed by a model of the principal components of the factor analysis. Four factors were extracted that explain 71.4% of the total variance. The factors-"general body voluminosity", "subcutaneous fat tissue", "longitudinal body dimensionality" and "upper body voluminosity"-were analyzed within the context of their appearance in different age-determined cohorts. The differences between cohorts (groups per ten years of age) were studied by the canonical discriminant analysis. The first two discriminant functions (describing mostly the variability of cohorts-96.11%) indicate a constant decrease of body and sitting height, and an increase of upper body voluminosity till the fourth age cohort, which is the most crucial one in the change of latent morphological structure. Results of the correct classification of cohort members show that only 48.45% of probands were correctly placed (the best classification determined was in the age between 46 and 55 years) indicating that in males, at least three different groups exist according to the specificity of morphological aging in human organisms. PMID- 9225506 TI - The effect of coagulation parameters on the placental respiratory and nutritive function in women having chronic hypertension with superimposed preeclampsia. AB - The aim of this study was to explore the relationship between coagulation and fibrinolytic system parameters with nutritive and respiratory placental function. We have analysed 79 pregnant women, of which 39 with severe preeclampsia (index group) and 41 healthy pregnant women. When comparing the study groups in third trimester, significantly lower platelet counts, fibrinogen values and antithrombin III values have been found in the index group compared to the control group. Factor VII levels were not found to be significantly different. The control group revealed significantly higher levels of coagulation factors II, V and VIII, compared to the index group. The increase of FDP, reduction of fibrinogen and increase of fibrinolysis in index group, when compared to the control group of healthy pregnant women, are the reflection of the intravascular fibrin deposition that leads to the described coagulation changes and consecutively to the foetal growth retardation. Indirect evidence are the correlation between newborns' weight and fibrinogen levels/fibrinolytic activity. PMID- 9225507 TI - Anthropometric and quantitative EMG status of femoral quadriceps before and after conventional kinesitherapy with and without magnetotherapy. AB - The frequency of femoral quadriceps muscle hypotrophy has become a significant therapeutic problem. Efforts are being made to improve the standard scheme of kinesitherapeutic treatment by using additional more effective therapeutic methods. Beside kinesitherapy, the authors have used magnetotherapy in 30 of the 60 patients. The total of 60 patients, both sexes, similar age groups and intensity of hypotrophy, were included in the study. They were divided into groups A and B, the experimental and the control one (30 patients each). The treatment was scheduled for the usual 5-6 weeks. Electromyographic quantitative analysis was used to check-up the treatment results achieved after 5 and 6 weeks of treatment period. Analysis of results has confirmed the assumption that magnetotherapy may yield better and faster treatment results, disappearance of pain and decreased risk of complications. The same results were obtained in the experimental group, only one week earlier than in the control group. The EMG quantitative analysis has not proved sufficiently reliable and objective method in the assessment of real condition of the muscle and effects of treatment. PMID- 9225508 TI - Deletion screening of the Duchenne/Becker muscular dystrophy gene in Croatian population. AB - The dystrophin gene deletion in 53 Duchenne and 21 Becker muscular dystrophy (DMD/BMD) male patients was analyzed by DNA test using multiplex polymerize chain reaction (M-PCR) in Croatian population. The overall percentage of deletion cases observed was 50%; 61% (53/32) for DMD and 38% (21/8) for BMD. The number of deleted exons was variable, but generally DMD deletions involving single-exon 19, 44, 50, 51 and larger exon deletions 3-6, 4-12, 4-17, 8-13, 12-13, 12-19, 48-50, 50-51, 50-52, 51-52 were more frequent. Eight patients with BMD had deletions exon 45-47, 45-48, and exon 3. The results obtained in the present study showed location of breakpoints in the dystrophin gene, and pointed to variability of deletion patterns in Croatian population among different European populations. PMID- 9225509 TI - A prospective study of S-T segment depression in the electrocardiogram and mortality in the population. AB - A prospective study was carried out of all degrees of horizontal or descending depression of the S-T segment in the electrocardiogram (more than 1 mm or 0.1 mV, 0.5-0.9 mm or 0.05-0.05 mV and up to 0.5 mm or 0.05 mV) in a sample of 2414 subjects of both sexes, aged 35-54 years, (1326 females and 1088 males) in six Croatian regions on three occasions during a 13-year period: 1969, 1972 and 1982, according to the Minnesota code. S-T segment depression in the ECG was found during the first examination in 10.69% of females and 4.13% of males; during the second examination in 12.66% females and 6.24% males, and in the third examination in 19.06% females and 12.12% males. S-T segment depression of up to 0.5 mm was dominant and twice as frequent in females than in males, and the difference was significant. S-T segment in the ECG was also analyzed in a sample of 239 subjects (141 males and 98 females) who died during the period between the second and third examination, and for whom ECGs had been recorded in 1969 and 1972. In the group of females who died during the period between the first and second examination, 16.32% had all degrees of S-T segment depression, while in the group of surviving females this amounted to 10.6%. In the group of deceased males this finding was found in 4.24% of deceased and 4.13% of surviving males. In the group of females who died between the second and third examination, S-T segment depression was found in 18.36% of deceased and 12.66% of surviving females, and 10.62% in deceased males and 6.24% in surviving males (depression of 1 mm or deeper and 0.5-0.9 mm was found in 7.08% of deceased and 1.28% surviving males). The relative risk of mortality in middle-aged females with S-T segment depression of 0.5 mm or deeper, was 3.65 times higher than in females without such ECG changes, while the relative risk of mortality in males of the same age with the same finding was 5.85 times higher than in those without such ECG changes. PMID- 9225510 TI - Comparative study of mitral annular calcification (MAC) with cardiac arrhythmias in dialysis patients. AB - Cardiac arrhythmias and myocardial malfunction are very frequent in uremic patients. The pathogenesis and etiology of arrhythmias are very complex and still unknown. The sedimentation of calcium salt in myocardial structures is one of the reasons for emergence of cardiac arrhythmias (AV conduction defects, ectopic arrhythmias). The appearance of mitral annular calcification (MAC), as the expression of the speed up process of atherosclerosis, was noted in younger uremic patients especially during hemodialysis. The aim of our research was to compare the incidence of MAC and cardiac arrhythmias in patients on hemodialysis. Our study included 40 patients, 24 male and 16 female, in the age between 20 and 60. Patients were mostly from Zagreb and the Counties of Zagreb (35%), Karlovac (10%), Slavonski Brod (7.5%), Varazdin (5%) and Pozega (5%). All 40 patients received 24 hours of Holter monitoring and 2-D echocardiography of M-mode. The patients were divided in two groups: I MAC+ (N = 23) and II MAC- (N = 17). Frequency of cardiac arrhythmias in group I was: atrial fibrillation N = 0; conduction defects N = 2 (1%); ventricularectopy Lown grade 3-5 N = 15 (65%); supraventricular ectopy N = 8 (34%), while the frequency of cardiac arrhythmias in group II was: atrial fibrillation N = 0; conduction defects N = 0; ventricular ectopy Lown grade 3-5 N = 6 (35%), supraventricular ectopy N = 6 (35%). During statistical processing the significant connection of MAC+ and frequency of cardiac arrhythmias was noticed. For both groups we have not noticed statistical significance in cardiac arrhythmia compared to electrolytes, risk factors PTH, and age. The time of hemodialysis treatment is one of possible factors for incidence of cardiac arrhythmias influenced by MAC. We noticed statistically significant (p < 0.05) difference of rhythm disorders between group I and group II especially for the ventricular ectopic activity, the frequency of which was higher in group I than in group II. MAC has probably significant role in dialysis patients for the development of cardiac arrhythmias within the framework of series of complicated multifactorial patogenetic mechanisms. PMID- 9225511 TI - Holistic approach to analysis of medical data: vulvar cancer. AB - This paper continues the series of studies introducing holistic approach to analysis of clinical data. Namely, besides the information regarding his/her disease, each hospitalized cancer patient also provides the variety of data regarding his/her psychological, cultural, social, economical, genetic, constitutional and medical background. The aim of this study was to introduce a holistic approach to analysis of medical data, in this case clinical data regarding cancer of the vulva. Such approach requires the collection of data regarding different aspects of the cancer patients, and after the satisfactory sample size is obtained (which should be at least five times greater than the number of examined patient characteristics), the performance of factor analysis. In this study, the authors have processed the data regarding 25 characteristics of all 755 vulvar cancer patients treated between 1938 and 1990 at the Department for Gynecological Oncology of the University Hospital for Gynecology and Obstetrics, Zagreb, Croatia. In factor analysis, the principal components were rotated after the initial extraction (the authors recommended the use of oblimin rotation) in order to obtain better ground for interpretation of the obtained results. The next step in this approach was the stepwise exclusion of characteristics with smallest commonality according to Kaiser-Meyer-Olkin criteria, and retaining the characteristics and components with the most significant impact on the explained system variance. When the number of principal components and initial analyzed characteristics was reduced to 3-4 and 7-10, respectively, the ultimate interpretations and conclusions were made. This approach outlined some clusters of correlations between medical data which are difficult to identify using other statistical procedures, primarily the impacts of various socioeconomic and hereditary-constitutional variables on overall survival. PMID- 9225512 TI - Holistic approach to analysis of medical data: cancer of the corpus uteri. AB - Besides the information regarding his/her disease, each hospitalized cancer patient also provides the variety of data regarding his/her psychological, cultural, social, economical, genetic, constitutional and medical background. The aim of this study was to introduce a holistic approach to analysis of medical data, in this case clinical data regarding cancer of the corpus uteri. Such approach requires the collection of data regarding different aspects of the cancer patient, and after the satisfactory sample size is obtained (which should be at least five times greater than the number of examined patient characteristics), the performance of factor analysis. In this study, the authors have processed the data regarding 25 characteristics of 928 corpus uteri cancer patients treated between 1980 and 1990 at the Department for Gynecological Oncology of the University Hospital for Gynecology and Obstetrics, Zagreb, Croatia. In factor analysis, the principal components were rotated after the initial extraction (the authors recommended the use of oblimin rotation) in order to obtain better ground for interpretation of the obtained results. The next step in this approach was the stepwise exclusion of characteristics with smallest communalities according to Kaiser-Meyer-Olkin criteria, and retaining the characteristics and components with the most significant impact on the explained system variance. When the number of principal components and initial analyzed characteristics was reduced to 3-4 and 7-10, respectively, the ultimate interpretations and conclusions were made. This approach outlined some clusters of correlations between medical data which are difficult to identify using other statistical procedures, primarily the impacts of various socioeconomic and hereditary-constitutional variables on overall survival. PMID- 9225513 TI - Estimation of radioactive calcium absorption based on expanding Ca-exchangeable space. AB - Anthropological interest in calcium metabolism is partially connected with the genetic basis of skeletal metabolism that could provide additional optimism for people suffering from osteoporosis. The present investigation is aimed at deal with a of model describing Ca intestinal absorption, suitable for other descriptions currently investigated in anthropology (migration and gene flow). The purpose of this investigation was to establish features of the reflection of the Ca isotopic absorption tests in serum using the concept of an expanding Ca space and to assess the reliability of the estimation of absorption (ABS) from one serum sample. Double tracer method, using 85Sr as the iv tracer, was used as the referent. The study included 100 subjects (49 females, 51 males) of which 34 were considered healthy. The mean value of ABS was 68 (+/-17.45 SD) % of dose. Serum radioactivity during the 24 hours following oral administration of 47Ca (together with 100 mg elemental Ca), o.s(t), from five blood samples was determined. The relationship between o.s(t) and ABS was found to be connected to an expanding Ca-space, V(t): ABS = o.s(t)V(t) = 26 + 11.8t0.35 o.s(t). After the absorption process was finished, an approximate value of V(t) is (100/S1)(t-x)b. The constants "S1" and "b" describe power function of iv s(t), whereas parameter "x" represents delay of V(t) Ca-space at oral administration in comparison to the iv administration. Statistical models selected on the basis of their low values of the SEE (from 8.71 to 8.90% of dose) include body surface index or body weight index (BW0.425). The CV expressed as (SEE/mean x ABS(Ca/Sr) x 100 were about 13% and the observed greatest difference between the estimated and measured ABS(Ca/Sr) was 20% of dose. The authors believe that models presented permit, to a limited extent, comparison of results obtained by different procedures. The results, according to variability of the sample of subjects, permit judgment on upper limits of error at estimation of absorption from one blood sample radioactivity. PMID- 9225514 TI - Some anatomical and anthropological measures of mandibular ramus in our population. AB - Beside the already analysed bioanthropometric characteristics of the lower jaw, additional measurement of parameters insufficiently discussed in reference literature were performed in our study. On the basis of the obtained data it was possible to define relevant biometrical relations in the region of the lower jaw branch and anatomical regions that are especially important to the use of conduction anaesthesia. All measurements were defined on the basis of anteroposterior (horizontal) and vertical directions. The relation between different anatomical structures of the lower jaw interior branch and foramen mandibulae served as a focal point. When related to anteroposterior plane the mandibular foramen was located precisely in the middle of the distance between crista temporalis and posterior ramus ridge. Viewed in vertical direction the lowest point of mandibular opening was slightly closer to the mandibular angle than to incisure of mandible, meaning that in the majority of macerated skulls mandibular foramen had a relatively low position. PMID- 9225515 TI - Comparative review of gnathometric characteristics in total dentures and eugnathic subjects. AB - The problem of determining the position and lining of the teeth in total dentures is presented by a comparative examination of gnathometric variables in eugnathic subjects and in those wearing total dentures. The material consisted of 51 plaster casts of eugnathic subjects and 49 subjects wearing total dentures. The following were examined: upper total number of incisors (T1), anterior width (AW), posterior width (PW), anterior length (AL) and posterior length (PL) of models. Analysis of the investigated variables was carried out by means of sliding calipers MECANIC Type 6901 (IVOCLAR, Lichtenstein) with a scale of 0-130 mm, and reading accuracy 97.5%. A three-dimensional pair of compasses according to Korhaus (Dentaurum, Germany) was used to measure the anterior and posterior lengths of models, with a scale 10-60 mm and reading accuracy 95%. No significant difference was observed between the eugnathic subjects and subjects wearing total dentures (p > 0.05). The results of this comparative gnathometric analysis of examined variables (Tl, AW, PW, AL and PL) are a contribution to the rules when lining anterior and posterior teeth in total dentures. PMID- 9225516 TI - Effect of unilateral function on craniofacial growth. AB - The present study is aimed at experimental research of the effects of functional matrices upon the formation of functional units. The sample consisted of 15 experimental dogs (five dogs per each experiment) and five controls. Three experiments were carried out: neurectomy of the right mandibular nerve, circumvention of the right nostril and enucleation of the right eyeball. The results showed no statistically significant difference between age and animal sacrifice time interval. The controls manifested the significant differences in the length, breadth and height of the upper mandibular body consistent with their natural asymmetry. Neurectomy of the right mandibular nerve did not manifest statistically significant differences whereas the sample was too small, animal sacrifice time interval too short and passive movements of that mandibular side were still present after the operation. Circumvention of the right nostril revealed in the nasal opening breadth, which for the very nature of the operation and only a partial exclusion of the right side function could have been anticipated. Enucleation of the right eyeball eliminated the visual function in the right face causing changes in the midface, i. e. in the breadth and height of the nostril. The findings confirmed the initial assumption on the correlation between the removal of potential growth factors and increase of the opposite face parameters. The changes were most prominent in the mid of the head which can be related to its developmental and structural features. PMID- 9225517 TI - The use of visual evoked potentials to follow-up prisoners of war after release from detention camps. AB - Visual evoked potentials were examined in released prisoners of war (POWs) in order to evaluate the extent of neurological impairment after imprisonment in Serbian detention camps. On two occasions visual evoked potentials were determined in a group of 11 released prisoners of war (POWs): 157 days and 379 days after release from detention camp. During the first examination no significant differences were found in VEP parameters between the right and left eye. However, during the second examination significant differences were found in the latencies of waves P100 and N145, statistically significantly prolonged latencies of the P100 wave and significantly greater amplitudes of waves P50 and N75. The results can be regarded as progression of the VEP changes in the released POWs. It is hypothesized that these changes are a result of a demyelination process, caused by the altered immunological status of the POWs during posttraumatic stress syndrome. PMID- 9225518 TI - Psychical difficulties in former prisoners of detention camps. AB - The war imprisonment is a traumatic experience which is generally considered to have a potential to cause various psychical difficulties, in particular the posttraumatic stress disorder (PTSD). During the aggression on Croatia, several thousands of Croatian soldiers and civilians were held in Serbian detention camps where they were tortured to extent of extreme stress. In this paper, the authors researched psychical effects of stress in former war prisoners. Examinees were observed in three separate groups. The first group was formed of all former prisoners of war (a total of 1458) that went through several medical examinations, including psychiatric one, after their release from detention camps. Examinations were performed at the University Clinic for Infectious Diseases "Dr. Fran Mihaljevic" in Zagreb, in period from November 1991 to September 1992. The second group consisted of 82 former prisoners randomly chosen from a total of 735 prisoners released from the "Sremska Mitrovica" camp in August 1992. The third group contained 37 prisoners from "Manjaca" camp out of 100 invited to the control examination 6 months after their release. A classic psychiatric diagnostic interview was performed in all of the examinees immediately after their release, and in the 2nd and the 3rd group the modified Watson's PTSD questionnaire was also used in addition. In the 2nd group, prisoners were questioned immediately after they were released. Using classical psychiatric interview, a specific psychiatric diagnosis could have been established in 20% cases. Psychiatric symptoms were observed in 30-40% examinees (in 36% of former "Manjaca" camp prisoners). Through the use of Watson's questionnaire, a PTSD diagnosis was established in 85.7% (70 out of 82) prisoners of the 2nd group, and in 27% of the 3rd investigated group of prisoners. Former prisoners examined after release (the 2nd group) showed significantly higher prevalence of PTSD symptoms. All PSTD symptoms were found in more than 50% cases of the 2nd group, while in the 3rd group none of the symptoms were found in more then half of examinees. Results are discussed and one among the direct conclusions is that former prisoners of war, expecting their problems to disappear spontaneously, are unwilling to seek for psychiatric help. PMID- 9225519 TI - Changing the stereotypes of "counsellors". AB - Stereotypes, as a relatively inflexible form of cognitive organization, serve in the organization of personal experience. They are relatively difficult to change and take a relatively long time. In our study we have tried to examine whether some traditional stereotypes of "counsellors" have altered during more than three years of work by members of the Croatian Psychiatrists Association with displaced persons in a camp in Zagreb. The statistical analysis of the information indicated that stereotypes have changed and in such a way that positive stereotypes have become more positive while the negative have become less negative among those displaced persons who had continued counseling from psychiatrists, psychologists and social workers from our Association. In the long term, these results can be considered even more positive than the actual process of psychological assistance because the change in stereotypes is likely to be reflected in a larger number of future clients and through future generations. PMID- 9225520 TI - Doctor's oath to secrecy and psychiatric patient. AB - Doctor's oath to secrecy is in the basics of patient's trust to doctor, which is a prerequisite of successful medical treatment. The authors tried to find the answers to following questions: a) what is the attitude of psychiatric patients, people who ask for psychiatric help, towards revealing facts about their psychical problems, b) how well are they informed about doctor's oath to secrecy and to what extent does their willingness reach in asking compensation on the court in case their secret was revealed, c) what is the frequency of the so called "institutionalized" revealing of secrets. The research was performed on the sample of 100 male psychiatric patients hospitalized at the University Clinic for Psychiatry and additional 100 persons who asked for help using phone service of Center for crisis conditions at the same clinic. The obtained data showed that 41% of hospitalized psychiatric patients wanted to hide their psychical problems, and considerably higher percentage (74%) of patients expect the doctor's discretion about the problem, although the most of them are not convinced that there will actually be one. Not a single patient has expressed willingness to ask for compensation on the court in case of doctor's breaking the oath to secrecy. About the half of the patients are informed about doctor's oath to secrecy (42%), but almost all of them (39% of cases in our total sample) consider it normal that the information about their illness is given to some institution outside their family. Persons that ask for help by phone want to remain anonymous in 43% of cases, and in almost same percentage (41%) they believe that their secrecy is guaranteed. In the concluding paragraphs, the need for constant consideration and actualization of problem of doctor's oath to secrecy in the complex situation of global social and medical progress is stressed. PMID- 9225521 TI - Investigation of the speed of reaction on external stimulus in schizophrenic psychosis. AB - In 30 schizophrenic examinees, the latention time was measured. This time is referred to as an interval between the start of the stimulus and the response to the stimulus in the skin-galvanic reflex. Elementary stimulation has been applied, using device's timer tone and clapping of hands, which should simulate and associate the thunderclap. The intensity of psychosis was measured according to the Metric scale of psychotic behavior by Rogina, while the intensity of anxiety was measured by psychological tests: Rorschach's psycho-diagnostic test and Spillberger's questionnaire for anxiety. The reaction to the stimulus and latention time were registered using polygraph unit in order to record skin galvanic reflex. The research was performed at two separate time points: prior to the therapy with derivatives of the phenothiazine group (the experimental examination group), and 25 days after the therapy (control group). The research has shown that the latention time in schizophrenic examinees does not significantly differ from the corresponding time in healthy controls, and it averages 2.30 seconds. Furthermore, no statistically significant difference in latention time before and after the therapy was observed. However, before the therapy started, i.e. in experimental group," the examinees who were psychotic to a greater extent have shown longer latention than those less psychotic. Additional finding was that the examinees from experimental group who were more anxious according to psychological tests have also shown longer latention time. After the therapy, the reaction to the external stimulus was stronger, which was expressed in increased reaction amplitude in skin-galvanic reflex. The latention time was prolonged, especially in case of examinees that were psychotic to a smaller extent before the therapy. We can conclude that so-called transformed psychotic anxiety was replaced after the therapy with a "new" anxiety-existential fear, i.e. the stronger anxious expectation and confusion appeared because the anxiety in its "free-floating" form remained very high. After the therapy, psychotic protection failed to appear and the reaction characteristics are a consequence of the delaying the confrontation with reality, which was registered as a prolonged latention time, i.e. response to external stimulus. PMID- 9225522 TI - The adolescents assessment of family functioning. AB - The aim of this study was to find out in what way adolescents assess their families and family functioning. An 34-item scale in self-report format was administered to 154 high school students and 34 patients in diagnostic procedure before entering psychotherapy in order to explore basic elements of family functioning (structure, affect, communication; behaviour control, ethic values transmission, intimacy and idealization, and elements of family dysfunction). The results showed that adolescents in general assessed their families as growth and individuation supportive. The adolescents in the clinical sample presented more dissatisfaction with their families and elements of family dysfunctions were more emphasized. PMID- 9225523 TI - Eating behaviour, weight status and depressive feelings in female adolescents. AB - The aim of this study was to examine the attitudes towards food and eating, body image, mood, feelings and relationships in family and peer group in female adolescents with varying weight status in an attempt to explore whether a relationship between emotional difficulties and body weight could be found. The data on use of 81 item measure of emotional difficulties and behavioural symptoms related to eating disorders and dieting in female adolescents aged 15 to 19 years and group of eating disorder patients are reported. The findings suggest that the adolescents who by self-reported weight value appear to be relatively overweight and eating disorder patients score significantly higher on a body dissatisfaction subscale of the applied questionnaire than normal weight and underweight adolescents. Only in the clinical sample of eating disorder patients, however, difficulties in dealing with depressive feelings and fear of poor impulse control were present. PMID- 9225524 TI - Continuity and change reflected in synchronic and diachronic linguistic variation of Middle Dalmatia. AB - The present paper analyses the geographical patterning of the two main dialects of the Croatian language in contemporary Middle Dalmatia, using language data as an indicator of population change by migration, with its wider cultural implications based on acculturation processes. It is shown that the synchronic evidence of the dialect and sub-dialect patterns of this area reflects more extensive interrelationships between different cultural substrata in terms of both little (local) and great (Islamic and Latin) traditions. PMID- 9225525 TI - Scientometric analysis of anthropology in the Republic of Croatia for the period of 1980-1996. AB - Anthropologists from the Republic of Croatia have published 254 scientific papers in the period from 1980-1996, that are included in the secondary publication Social Science Citation Index. Scientists working in the scientific subfield anthropology participate with approximately 2% in the overall scientific output of the Republic of Croatia. Thirty-six international articles were published (14.2% of the total number), while the rest of 218 papers were published solely by domestic authors. An average anthropological paper is published by 3.06 authors, and approximately one-third of all articles by a single author. The major part of scientific papers (237 articles or 93.3%), Croatian anthropologists have published in a domestic primary scientific journal Collegium Antropologicum. All scientific papers together obtained 380 citations or 1.5 citations per article. The citation of articles is approximately 60% above the expected average for the respective journals. Published international papers had 6.6 citations, while articles by domestic authors had 0.65 citation per paper. Anthropological scientific papers obtained 154 independent citations and participate with 40.5% in the total number of citations. In the first five years after publishing, 166 articles (65.4% of the total number) were not cited, while the world's average for the scientific subfield anthropology was greater, 79.5% uncited articles. Only 19.4% of international papers and 72.9% of domestic papers were not cited in this five-year period. Based on scientometric indicators of a scientific output, that is, the number of published papers, partial scientific contribution, i.e., partial authorship, and scientific influence, i.e. number of citations, a method for the evaluation of scientific papers and their authors has been suggested in this paper. PMID- 9225526 TI - Analysis of the quantitative dermatoglyphics of the digito-palmar complex in patients with multiple sclerosis. AB - Recent studies on the etiopathogenesis of multiple sclerosis (MS) all point out that there is a polygenetical predisposition for this illness. The so called "MS Trait" determines the reactivity of the immunological system upon ecological factors. The development of the glyphological science and the study of the characteristics of the digito-palmar dermatoglyphic complex (for which it was established that they are polygenetically determined characteristics) all enable a better insight into the genetic development during early embriogenesis. The aim of this study was to estimate certain differences in the dermatoglyphics of digito-palmar complexes between the group with multiple sclerosis and the comparable, phenotypically healthy groups of both sexes. This study is based on the analysis of 18 quantitative characteristics of the digito-palmar complex in 125 patients with multiple sclerosis (41 males and 84 females) in comparison to a group of 400 phenotypically healthy patients (200 males and 200 females). The conducted analysis pointed towards a statistically significant decrease of the number of digital and palmar ridges, as well as with lower values of atd angles in a group of MS patients of both sexes. The main discriminators were the characteristic palmar dermatoglyphics with the possibility that the discriminate analysis classifies over 80% of the examinees which exceeds the statistical significance. The results of this study suggest a possible discrimination of patients with MS and the phenotypically health population through the analysis of the dermatoglyphic status, and therefore the possibility that multiple sclerosis is genetically predisposed disease. PMID- 9225527 TI - Changes in vitamin E concentration after surgery and anesthesia. AB - The aim of this randomized study was to examine changes in vitamin E concentration in female subjects (age 30-60, ASA I) after cholecystectomy and halothane (N = 16) or isoflurane (N = 16) anaesthesia. Vitamin E concentration was measured two days before, and then one, five and twenty-four hours and four days after surgery. High-pressure liquid chromatography was used for its determination. Simultaneously activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma glutamyltransferase (GGT) were determined. STATISTICAL ANALYSIS: ANOVA, Tukay HSD test. The research has been accepted by the Drugs Committee of the Karlovac County Hospital. Preoperative vitamin E concentrations in the halothane group were 8.69 +/- 2.35 micrograms/L, median 8.67 micrograms/L and in the isoflurane group 9.43 +/- 2.4 micrograms/L, median 9.08 micrograms/L. Statistically lower vitamin E concentrations compared with preoperative values were noted one hour (P < 0.05), 5 hours (P < 0.01), 24 hours (P < 0.01), as well as 4 days (P < 0.01) after the operation. The lowest vitamin E concentrations were noted 24 hours after the operation with statistically insignificantly higher values in the isoflurane group (halothane group 5.98 +/- 2.08 micrograms/L, isoflurane group 6.58 +/- 1.51 micrograms/L). Analyzing enzyme (ALT, AST and GGT) pre- and postoperative values, no statistically significant differences between the investigated groups and during the time were observed. Statistically significant differences were found between individual measurement times, with no statistical significance of the differences between the halothane and isoflurane groups. It seems that neither the difference in halothane and isoflurane biotransformation nor their distinct effect on perfusion of some organs are the determining factors in post-operative changes in vitamin E concentration. PMID- 9225528 TI - Awarding of the Herder Award to Dunja Rihtman-Augustin. PMID- 9225531 TI - Bringing an evidence-base to dentistry. PMID- 9225532 TI - Oral health education materials; whose business is it anyway? PMID- 9225533 TI - Patient satisfaction with a consultation at a cranio-facial pain unit. AB - OBJECTIVE: Patient satisfaction is an important component of the evaluation of the quality of health care and has also been linked to therapeutic outcomes. The objective of this paper was to assess patient satisfaction with an initial consultation at a cranio-facial pain unit located in a large hospital in a major metropolitan centre. DESIGN: A case series design was used. The study population consisted of new patients referred to the unit. One month before their initial appointment patients were mailed an oro-facial pain questionnaire. Immediately following the consultation they were mailed a satisfaction questionnaire which contained a modified version of the Dental Visit Satisfaction Scale. RESULTS: Oro facial pain data were collected from 121 patients and 78 of these returned a completed satisfaction questionnaire. While levels of satisfaction with the consultation were high overall, specific questions about consultation processes and outcomes revealed some sources of dissatisfaction. Just over half reported dissatisfaction with communications and one fifth were dissatisfied with outcomes in terms of diagnosis and treatment. Satisfaction with communication was the only dimension of the consultation process to be associated with satisfaction with outcomes, and satisfaction with outcomes was the main predictor of satisfaction overall. CONCLUSIONS: Although the relationships between patients and practitioners at the unit were favourable, improvements in communications may be necessary if the full benefits of the consultation are to be realised. This can be difficult with this patient population whose conditions are complex and often poorly understood. PMID- 9225534 TI - A clinical trial of two fluoride dentifrices in an area of low caries prevalence. AB - OBJECTIVE: The study was designed to test the hypothesis that a dentifrice with fluoride at the same concentration (1000ppm) from two sources, ie NaF and NaMFP, would provide a greater treatment effect than one with NaMFP alone. BASIC RESEARCH DESIGN: A double blind clinical trial with random assignment of children to one of two groups was carried out for three years. The two trial groups were similar at the outset in respect to those variables which might otherwise have affected the outcome, including age and gender, with means per subject of 98.4 sound surfaces and 2.2 decayed and filled surfaces in each group initially. CLINICAL SETTING: Secondary schools in the Isle of Wight, UK, an area of diminished caries experience. PARTICIPANTS: One thousand six hundred and thirty three children aged initially 10-12 years. INTERVENTIONS: A test dentifrice containing 500ppm NaF plus 500ppm NaMFP, and a standard active control product containing 1000ppm NaMFP. Products were used in the home. OUTCOME MEASURES: Increment of DF teeth and surfaces measured over 36 months. RESULTS: After three years, mean approximal surface increments were 3.6 new DFS in the control group and 3.1 in the test group, a difference 13 per cent (P < 0.05). Thirty-four per cent of the subjects were caries free at the outset. In the 1075 subjects with caries at the outset, the total mean increment on all surfaces was 7.2 new DFS in the control group and 6.4 new DFS in the test group, a difference of 11 per cent (P < 0.05). However, those subjects with initial caries had approximal surface increments of 4.8 new DFS in the control group and 4.0 new DFS in the test group, a difference of 16 per cent (P < 0.01). Included separately along with the conventional rubric were enamel white spots on which no differential treatment effect was observed. CONCLUSIONS: Under the conditions of this study, the regular use of a dentifrice containing 1000ppm fluoride from two sources provided a significantly greater treatment effect than one with fluoride from a single source. PMID- 9225536 TI - Trends in toothbrushing frequency among Finnish adolescents between 1977 and 1995. AB - OBJECTIVE: To analyse trends in development of the toothbrushing frequency of Finnish adolescents and the socio-economic factors associated with these trends between 1977 and 1995. DESIGN: The data were collected as part of a nation-wide research programme, the Adolescent Health and Lifestyle Survey, which started in 1977. Since then a 12-page questionnaire has been sent every other year. Dental health behaviour was studied from the outset. SUBJECTS: The sample represented 12 , 14-, 16- and 18-year-old children and adolescents in Finland. The sample size varied between 3,205-10,626, making a total of 66,687 participants. OUTCOME MEASURES: The recommended toothbrushing frequency, twice-a-day, was studied. The socio-economic factors included age, gender, self-assessed school performance, level of education, socio-economic status of the householder, and socio-economic category of the residential area. RESULTS: Among boys, daily toothbrushing increased from 1977 to 1995, but among girls it remained stable. Among boys, the prevalence of twice-a-day toothbrushing frequency varied from 13 per cent to 25 per cent between the ages of 12 and 18 years, and among girls from 32 per cent to 60 per cent, respectively. Among 12- to 14-year-old boys, the socio-economic differences almost disappeared. There were no changes among 12- to 14-year-old girls but there was an unexpected declining trend in toothbrushing among 16- to 18-year-old girls. Apparently further improvement in the toothbrushing frequency of girls had stopped. CONCLUSIONS: Although there was a clear trend towards improvement of toothbrushing frequency among boys, their toothbrushing frequency still lagged far behind that of girls. PMID- 9225535 TI - Development of an oral health indicator in infants. AB - OBJECTIVE: To develop a simple predictive indicator for 1.5-year-old infants' oral health as an aid for maternal preventive efforts. DESIGN: The development of an indicator was based on the relationship between life style factors of the 1.5 year-old infants and the caries incidence of the same children at three years of age. Data gained at examination of the 3-year-old subjects were subjected to analysis with the caries-free information on the same subjects at 1.5 years. SETTING: A community health centre in Hiroshima Japan. SUBJECTS: 1575 children received an oral check-up at 3 years of age at a community health centre. OUTCOME MEASURES: The category scores of the six items which were extracted retrospectively from life style factors at 1.5 years affecting caries prevalence at 3 years of age were quantified and transformed into integral numbers so as to make up a total of 100. The sum of the six transformed scores was named an Infants Dental Index (IDI). RESULTS: The IDI score was associated significantly with the dmft score at 3 years of age (r = -0.18 P < 0.001). When the subjects were classified into four groups with respect to the IDI score these four groups were significantly related to the caries onset between 1.5 and 3 years of age (chi 2 = 36.74, P < 0.001 CONCLUSIONS: The IDI developed in this study appeared to be valid so that it could be applied in the field of the community dental health to identity higher risk infants and direct more effective health education to mothers. PMID- 9225537 TI - An emergency dental service for students: 4-year findings. AB - OBJECTIVE: To describe the arrangements for the provision of emergency dental services for students at the University of Manchester and to report data collected during the first four and a half years of the student emergency dental services (SEDS) unit based at the University Dental Hospital of Manchester. METHOD: Data pertaining to every student attending SEDS since its inception were collected by means of questionnaire including provision to record diagnoses, treatment needs and the emergency care provided. RESULTS: The incidence of dental emergencies within the student population served by SEDS has been found to be 39 emergencies per 1000 students per annum, with the service being most heavily used by overseas students. Caries, pulpal pathology and failed restorations account for 46 per cent of the presenting emergencies, with pericoronitis (19 per cent) and other emergencies of periodontal origin (14 per cent) being common place. CONCLUSION: It is concluded that a student emergency dental service may be found to be an important element of student medical and related welfare services. PMID- 9225538 TI - A pilot assessment of alternative methods of quantifying dental pain with particular reference to dentine hypersensitivity. AB - OBJECTIVE: The overall aim of this pilot study was to establish the usefulness and comparability of selected verbal and non-verbal methods in the quantification of sensory and affective aspects of dental pain associated with dentine hypersensitivity (DH). DESIGN: The assessment of dental pain was conducted during an eight week clinical study. Patients were asked to rate their perception of dental pain using selected methods of quantification following tactile (Yeaple Probe-an electronic pressure-sensitive probe) and evaporative (cold air from a dental air syringe) stimulation; together with an overall assessment of perception to daily stimuli (e.g., cold air/water, toothbrushing, sweet and sour foods). The assessment methods used to quantify pain arising from DH were a continuous visual analogue scale (VAS), a 0-10 numerical rating VAS scale (NRS), and a separate intensity verbal descriptor (IVD) and unpleasantness verbal descriptor (UVD) word scales. SETTING: A specialist department at a postgraduate dental institute and hospital in London, UK. SUBJECTS: Twenty-five adult patients (8M + 17F) with a mean age of 42.6 years (95 per cent C.I. 38.8 to 46.4 years) attending the department for a clinical study evaluating the efficacy of a desensitising toothpaste agreed to participate. OUTCOME MEASURES: The study compared a continuous visual analogue scale (VAS), a 0-10 numerical rating visual analogue scale (NRS), and a separate intensity (IVD) and unpleasantness verbal descriptor (UVD) scales to quantify sensory and affective aspects of pain. An unweighted moving average technique was used to construct graphs of the relative frequency of reported severity gradings over a range of 0-10. RESULTS: The results of the study indicated that cold air appeared to cause greater discomfort to the patient than tactile sensitivity, with the air intensity curve for both IVD and 0-10 VAS peaking at a severity score of 5 while continuous VAS peaked at a score of 3-4. All methods peaked at score 2 for tactile sensitivity. The UVD scale peaked at score 2-3 and again at 6 for air sensitivity, but conformed to the other scales by peaking at score 2 for tactile sensitivity. NRS and IVD scales therefore appeared to provide acceptable alternatives to continuous VAS, but the UVD scale, probably because of the imprecise nature of the words used in the scale, did not. CONCLUSIONS: This study partially confirms previous conclusions that both verbal and non-verbal techniques quantify sensory and affective aspects of pain. However, the imprecise nature of UVD words provided misleading information in terms of both accuracy and sensitivity (except at very low levels of discomfort), when assessing pain arising from dentine hypersensitivity. In view of the highly subjective data arising from studies of this nature, the use of a moving average technique may be considered a more pragmatic method of analysis. PMID- 9225539 TI - Multifactorial modelling for caries prediction in Jordanian university students. AB - OBJECTIVE: To construct a prediction model for caries experience in Jordanian university students using a number of explanatory risk factors as predictors. DESIGN: Data on salivary flow rate, buffering capacity, streptococci and lactobacilli counts, plaque accumulation, oral hygiene and between meal sugar intakes were tested as predictors of clinically and radiographically registered DMFS: Methods of analysis included correlation, then multiple regression, and finally dichotomisation of the DMFS data and application of discriminant analysis and logistic regression. The latter analyses were conducted in order to predict in which caries risk group an individual belonged rather than predicting (as with regression) their actual caries status. Two dichotomisation schemes were investigated; dichotomisation at the mean and at the 75th percentile. SETTING: The University of Jordan. PARTICIPANTS: A random sample of 180 university students (77 male and 103 female). OUTCOME MEASURES: Relationships were expressed as correlation coefficients, R2, and sensitivity, specificity and predictive values of the predictors, and also their validity and efficiency. RESULTS: The highest correlation coefficient achieved was 0.43 (P < 0.0001) between sugar containing snack intakes and DMFS. The predicted power of the fitted multiple regression model was low R2 = 0.38). Logistic regression with the DMFS data dichotomised at the 75th percentile indicated that the fitted caries model correctly identified 76 per cent of the subjects. Sensitivity and specificity values of the predictive battery were 80 per cent and 75 per cent respectively. CONCLUSIONS: The multifactorial aetiology of caries remains unclear and requires further research. In the meantime, well-documented preventive measures should be implemented for this and similar populations. PMID- 9225540 TI - Stages of change for sugar and fat reduction in an adolescent sample. AB - OBJECTIVES: To apply the stages of change model for sugar and fat intakes in a sample of adolescents and to assess the factors influencing young people's ability to change their eating patterns. BASIC RESEARCH DESIGN: Self complete questionnaires assessed young people's readiness to change both their sugar and fat intakes in a cross sectional study. PARTICIPANTS: The study sample consisted of 479 young people aged 13-14 years attending four mixed ability state secondary schools in Camden, North London. RESULTS: A sizeable proportion of the sample were either in the precontemplation or action stages for their sugar or fat consumption. There were significant differences between the males and females. Application of the stages of change model produced very similar results for both sugar and fat consumption. The main reason given for reducing sugar or fat intakes was a desire to improve appearance through losing weight. Direct health concerns were less of a concern. A range of social and structural factors were identified by the sample as having an influence over their ability to make future dietary changes. CONCLUSIONS: Future oral health promotion interventions designed to promote healthier eating practices amongst young people need to recognise the various stages of change young people may be in and develop appropriate measures to meet their needs. PMID- 9225541 TI - Accessing primary dental care in three London boroughs. AB - OBJECTIVE: To investigate and identify factors that may influence individuals' ability to access primary health care services. DESIGN: Cross-sectional study SETTING: Lambeth, Southwark and Lewisham. SUBJECTS: A 0.1 per cent random sample of people aged 18 years and over in the population of 626,621 people in the three boroughs. MAIN OUTCOME MEASURES: Demographic as well as psychosocial profile of the sample, perceptions of their dental health needs, accessed dental care. RESULTS: The results indicated that people believed that their oral health was good. The majority of the sample had attended for dental care within the previous year and were registered with a general dental practitioner. Accessing dental care was related to age, social class, borough of residence, dentate status and dental phobia status. In addition subjects experiencing problems with their teeth gained access to care more readily than others. This was related to social class. Accessing dental care was predicted by dental care being provided by a general dental practitioner, experiencing problems with teeth and not being dentally phobic. CONCLUSIONS: The findings suggest that psychosocial factors together with dental health status can act as determinants when accessing primary dental services. It is considered that family health services authorities should be aware of these influences when developing and monitoring dental health services, in order to make them responsive and sensitive to the needs of the people whom they serve. PMID- 9225543 TI - Coming of age: cultural diversity and mental health. PMID- 9225542 TI - Presidential address to the British Association for the Study of Community Dentistry, Reading, April 1997. PMID- 9225544 TI - Personality disorders and culture: contemporary clinical views (Part A). AB - This article reviews the basic concepts surrounding the clinical relationships between culture and personality disorders (PDs). Culture plays a significant role in the construction of self-concept and self-image, the egocentric/sociocentric dichotomy, and the determination of biases in the clinical study of PDs. Cultural contextualization is, therefore, crucial in the demarcation between normal and abnormal personalities. From a clinical perspective, culture has three roles vis a-vis the psychopathology of personality: (a) as an interpretive/explanatory tool; (b) as a pathogenic/pathoplastic agent; and (c) as a diagnostic/nosological factor. The first of two parts, this article examines the interpretive/explanatory and pathogenic/pathoplastic roles, substantiated by clinical examples gleaned from the existing literature. PMID- 9225545 TI - Psychological testing as a diagnostic and therapeutic tool in the treatment of traumatized Latin American and African refugees. AB - The use of psychological assessment, an underutilized tool, in connection with posttraumatic stress disorder (PTSD) is presented. Identification of PTSD in refugees from Latin America and Africa is usually difficult because it is compounded by the trauma of migration. Issues regarding diagnosis and treatment are discussed, and case examples are provided to illustrate specific clinical concerns. Disclosure of historical information to the clinician and validation of a history of trauma are addressed through the testing process and projective data patterns. PMID- 9225546 TI - Personal storytelling and the metaphor of belonging. AB - The author uses the idiom of autobiographical storytelling to explicate two important themes: How the complex phenomenon of 'belonging' results from the interactions of members of a nondominant group and those of a dominant group; and how the author himself has dealt with this phenomenon in his own developmental adaptation over the years. The concept of belonging is a cultural principle that should be thoroughly understood by mental health professionals if they are to deal effectively with the problem of cross-cultural adaptation. PMID- 9225547 TI - Psychopathology among Asian Americans: a model minority? AB - The prevalence of psychopathology among Asian Americans has been a source of debate. Some investigators believe that the prevalence rate is quite low, whereas others argue that it is fairly high. A review of the literature suggests that at this time, it is not possible to determine the specific rates of psychopathology. However, evidence does suggest that their rates of mental disorders are not extraordinarily low. Thus, public portrayals of Asian Americans as a well adjusted group do not reflect reality. Attempts to determine the exact prevalence rates have been hindered by characteristics of the Asian American population, particularly its relatively small size, heterogeneity, and rapid changes in demographics. It is suggested that aggregate research, in which different Asian American groups are combined, is important for policy considerations, broad cultural comparisons, and establishing baseline information. To advance scientific contributions and understanding, studies that examine the correlates and course of disorders within specific Asian American groups are necessary as well. PMID- 9225548 TI - Black women, work, stress, and perceived discrimination: the focused support group model as an intervention for stress reduction. AB - This exploratory study examined the use of two components (small and large groups) of a community-based intervention, the Focused Support Group (FSG) model, to alleviate employment-related stressors in Black women. Participants were assigned to small groups based on occupational status. Groups met for five weekly 3-hr sessions in didactic or small- and large-group formats. Two evaluations following the didactic session and the small and large group sessions elicited information on satisfaction with each of the formats, self-reported change in stress, awareness of interpersonal and sociopolitical issues affecting Black women in the labor force, assessing support networks, and usefulness of specific discussion topics to stress reduction. Results indicated the usefulness of the small- and large-group formats in reduction of self-reported stress and increases in personal and professional sources of support. Discussions on race and sex discrimination in the workplace were effective in overall stress reduction. The study highlights labor force participation as a potential source of stress for Black women, and supports the development of culture- and gender-appropriate community interventions as viable and cost-effective methods for stress reduction. PMID- 9225549 TI - Can psychotropic medications change ethnoculturally determined behavior? AB - A clinical case is presented in which the use of a prescribed psychotropic medication was associated with dramatic changes in ethnoculturally determined attitudes and behavior. These changes were first noticed by the patient and followed a month of improvement in the target symptoms for which the medication was prescribed. The hypothesis is advanced that changes in identity may occur only after the establishment and adjustment to a new neurohormonal equilibrium. Medication-induced personality changes may facilitate changes in ethnocultural identity. PMID- 9225550 TI - Personality disorders and culture: contemporary clinical views (Part B). AB - This article reviews the basic concepts surrounding the clinical relationships between culture and personality disorders (PDs). Part A of this article, which appeared in Cultural Diversity and Mental Health, Vol. 1, No. 1, pp 3-17 (1995), examined the interpretive/explanatory and pathogenic/ pathoplastic roles of culture. Herein, culture's role as a diagnostic/nosological factor is discussed through the use of measurement instruments and the cultural formulation included in DSM-IV (American Psychiatric Association, 1994). In addition to these three roles, some authors would also consider a therapeutic/protective function for cultured in PDs. Following a critique of the biological perspective, a research model based on the definition of the cultural profile and the estimation of the cultural distance between clinical examiners and populations is proposed. It is important to reject both biological reductionism and the extremes of cultural determinism, in order to better assess the intraethnic distribution of psychopathology, and interethnic variations represented by the notion of cultural relativism. PMID- 9225552 TI - From graduate student to ethnic researcher: a challenging journey. AB - The goal of this position article is to convey the message to junior faculty with diverse personal characteristics (i.e., female, minority, or majority ethnic scholars) that obstacles encountered in their academic experience may be the result of being perceived as a symbol of sociohistorical problems. For conveying this message, I will tell my story by referring to what I have experienced as transitional phases: (1) "becoming an international student," (2) "from ambassador to minority," (3) "the great insight, sharing our problems," and (4) "overcoming using coping strategies in academic life." Emphasis will be given to coping strategies that I have developed for facing challenging sociohistorical problems while protecting my psychological well-being. Although the literature is sparse, coping strategies are discussed using available literature and personal experiences including the ability: (1) to assume that most problems affecting diverse junior faculty are sociohistorical, (2) to acknowledge the importance of finding mentors for the psychological well-being of junior faculty, (3) to stop replying to stereotypic views and to use energy and time more productively, (4) to respond selectively to collegial and student feedback, (5) to proactively establish collaborative activities, and (6) to have clear career development objectives. I conclude by recognizing the need for junior faculty to assume an advocacy role for creating a new era of equality and change in academe and in multicultural America. Thus, junior faculty need to undergo a major insightful realization leading to an awakening experience portrayed by the phrase: It is not only me. PMID- 9225551 TI - The therapist of color and the white patient dyad: contradictions and recognitions. AB - The therapist of color and White patient dyad often involves contradictions and recognitions which are acknowledged through the specific processes and dynamics permeating this dyad. The relationship between self and other is frequently mediated through projection and identification. This article examines this unique interracial and interethnic therapeutic dyad emphasizing its clinical implications through the attribution of otherness, the use of colored screen projection, and the significance of power reversal. Special emphasis is given to the prevalent transferential and countertransferential reactions aided by clinical material. It is concluded that this therapeutic dyad provides a model for cross-cultural encounters where the resolution of contradictions can lead to the recognition of paradoxes, the acknowledgment of ambivalence, and the acceptance of disparate parts of the self. PMID- 9225553 TI - Beauty is in the soul of the beholder: psychological implications of beauty and African American women. AB - The criteria for beauty in the United States are primarily based on Caucasian European American, middle-class standards. African American women tend to vary greatly from these criteria. Though very few studies have been conducted on the body image of Black women in the United States, historically, the physical images portrayed of African American women in the United States have not been positive. Mental health practitioners must understand how these negative images may affect the body image and self-esteem of African American women. Therapeutic and community interventions are discussed. PMID- 9225554 TI - Coping with culture-based conflict: implications for counseling research and practice. AB - Interpersonal conflict related to sociocultural group membership was examined with a multicultural university sample. The Social Group Conflict Scale (SGCS), collective self-esteem (CSE), and Bradburn affect scale were administered to 248 university students. The current study attempted to replicate and extend the findings on social group-based conflict recently proposed by Dunbar, Sue, and Liu. Results indicated that 51% of the subjects reported encountering interpersonal conflict attributable to their social group memberships, with ethnicity being the most frequently attributed group category. Significant gender and ethnic differences were noted in coping approach employed in responding to the conflict event. The current findings are considered in regard to effectively assessing and responding to intercultural conflict for mental health practice. PMID- 9225555 TI - Brief symptom inventory scores of Asian, Asian-American, and European-American college students. AB - Brief Symptom Inventory scores of Asian, Asian-American, and European-American university students were examined. It was hypothesized that Asian-Americans with low-medium levels of acculturation would show levels of distress similar to those found with Cheng, Leong, and Geist's sample of international Asian college students, whereas Asian-American research participants with higher levels of acculturation would report levels of distress more similar to European-American college students. The results indicated support for the influence of level of acculturation on the reporting of distress on the BSI and further indicated the need for future research on Asian-Americans to examine gender effects in addition to acculturation. PMID- 9225556 TI - Grief in family and cultural context: learning from Latino families. AB - The following article offers an integrative theory of family development in sociocultural context, which critically examines the goodness of fit between the grief experiences of culturally diverse families and prevailing North American grief practices. This model suggests that many Latino families, even when they are themselves from a variety of national backgrounds with somewhat different sociopolitical histories and cultural practices, offer an approach to death and grief that can enhance developmental outcomes in family bereavement. These qualities include a focus on family and extended family relationships as developmental resources; an appreciation of the spiritual and psychological continuity between the living and the dead; and an appreciation of the need to keep working on relationships, even after a death, so as to create new, more optimal shared understandings of family past, present, and future. The bereavement story of the Ruiz family, who migrated from rural Puerto Rico to the United States, serves to illustrate the ways that integration of Latino family experience into mental health models of grief can help expand understanding and improve bereavement outcomes for all families. PMID- 9225557 TI - Either a paradigm shift or no mental measurement: the nonscience and the nonsense of The Bell Curve. PMID- 9225558 TI - Exile and professional identity: on going back to Cuba. AB - The author tells the story of her lifelong attempts to create a coherent, complex cultural identity from her family's multiple diaspora legacies, and the impact of these struggles on her personal and professional development. She emphasizes the intergenerational conflicts created by the sociopolitical circumstances of her generation's Cuban immigration experience, and progressive attempts to include her Cuban identity in her sense of self. An unexpected lesson in the politics and history of psychoanalysis, itself an immigrant movement that abandoned its social conscience to survive in exile, catalyzed a return to Cuba and a greater inclusion of its social values in her personal and professional lives. PMID- 9225559 TI - The origin and formulation of Chinese character: an introduction to confucianism and its influence on Chinese behavior patterns. AB - Confucianism has had a powerful influence on Chinese behavior and social structure for the past 2,000 years. Some of these influences produce roadblocks to conventional Western psychotherapy and counseling when used on Chinese Americans with traditional values. The Confucianism influence on the Chinese is described with suggestions for psychotherapists and counselors who have Chinese and Chinese American clients. PMID- 9225560 TI - Severity of disturbance among Asian American outpatients. AB - Although Asian Americans are low utilizers of mental health services, they may suffer from a greater degree of disturbance by the time they are accepted for services. In this study, thousands of Asian and White clients who utilized a large mental health system over a 5-year period were compared on three measures of severity of disturbance: severity of diagnosis, ratings of functioning, and presence of psychotic features. Results for all three indicators supported the hypothesis that Asian Americans show greater disturbance than do Whites. The findings provide convincing evidence of greater disturbance among Asian American clients. It is suggested that for cultural reasons, only the most severely disturbed Asian clients use mental health services. PMID- 9225561 TI - Intercountry adoption of Latin American children: the importance of early bilingual/bicultural services. AB - Because of major worldwide demographic changes, many Latin American children are frequently adopted in the U.S. This article presents an overview of the historical and contemporary circumstances and controversies surrounding intercountry adoptions (ICAs), and a review of possible risk factors for later child or adolescent maladjustment. Although a number of follow-up studies indicate a 70-80% positive outcome, some ICAs end in painful family-child disruptions. There is, therefore, a growing need for bilingual/bicultural mental health services to improve the initial adjustment process and to facilitate a positive long-term outcome. These services are especially needed when the child is older at arrival and the adopting family lives in a mostly homogeneous community. Suggested prearrival, arrival, and follow-up interventions with the family, the child, and the school are described. A case illustration of the interventions offered to a U.S. family and their recently adopted Latin American child is provided. PMID- 9225562 TI - The Simpson trial: lessons for mental health practitioners. AB - The author reflects on the emotional impact of the O.J. Simpson trial and verdict and contends that the issues and concerns generated by the trial are mental health issues. The lack of leadership of mental health professionals in helping people to deal with their reactions to these events is noted. From this experience, seven lessons for mental health professionals are proposed. In particular, a call is made for mental health practitioners who are committed to cultural diversity to become more involved in understanding and responding to racially charged, community-rupturing events like the Simpson trial and verdict. PMID- 9225563 TI - "Dichos" therapy group: a therapeutic use of Spanish language proverbs with hospitalized Spanish-speaking psychiatric patients. AB - Physical, emotional, and cultural barriers have resulted in the underutilization of mental health services by Hispanic/Latino individuals. Described in this article is a culturally sensitive intervention that has been developed based on existing elements within Hispanic/Latino culture. The therapeutic uses of dichos- Spanish language proverbs and refranes (sayings)--are explored in the treatment of a regressed population of Hispanic/Latino psychiatric inpatients. The dichos therapy group described in this article utilizes dichos to draw patients into discussions about a wide range of issues, and is able to do so where other efforts fail because of their cultural and familial relevance, vivid imagry, and the flexibility with which they can be used. This intervention effectively facilitates building rapport, decreasing defensiveness, enhancing motivation and participation in therapy, improving self-esteem, focusing attention, stimulating emotional exploration and articulation of feelings, and development of insight, and assists in exploring cultural values and identity. Clinical examples are provided to exemplify these therapeutic effects. PMID- 9225564 TI - Consciousness-raising groups as a multicultural awareness approach: an experience with counselor trainees. AB - This article discusses the usefulness of consciousness-raising (CR) groups in the training of multiculturally aware counselors, and provides a description of their use in a graduate course on multicultural and gender issues in counseling. The conceptual framework for this training experience is built on Young's (1992) analysis of oppression, Freire's (1971, 1973) theory of liberating pedagogy, and social constructionist philosophy. Course procedures and several CR activities are described; the training experience is evaluated in terms of student reactions to it and potential dilemmas an instructor might face in using this format for teaching. PMID- 9225565 TI - Ethnocultural factors in the development of an Asian American psychiatrist. AB - Despite rising numbers of Asian American psychiatric trainees, little has been written about the specific problems arising in training for members of this ethnic minority group. The authors discuss some of the difficulties for the Asian American psychiatric trainee, in relation to the stigma of mental illness and its impact on the trainee's decision to enter psychiatry, ethnic identity and stereotyping, psychotherapy supervision, and career opportunities. Specific vignettes will describe each of these situations and the internal conflicts they engender during training. The resolution of these conflicts will be described within a transference and countransference framework with the intent of providing a starting point for process-oriented supervision geared toward the development of a professional identity. Specific recommendations will be given for the educational and career development process for Asian American psychiatric trainees. PMID- 9225566 TI - Psychosocial risk and protective influences in Hawaiian adolescent psychopathology. AB - A large community sample of adolescents of a Native Hawaiian (Asian/ Pacific Islander) minority group was studied along with a small comparison group of non Hawaiians, for the relationship between psychopathology (as measured by standard symptom scales) and (a) perceived support from family and friends, and (b) discussing problems with others. Expected gender patterns for friend support but not for family support were found. The Hawaiian boys appeared atypical, reporting nearly equal family support as Hawaiian girls. Discussing problems with another person was correlated with lower anxiety and depression scores but not aggression and substance abuse scores. It is concluded that gender and cultural factors influence symptom prevalence and severity as well as the impact of psychosocial risk factors. PMID- 9225567 TI - The influence of race and gender on depressive and substance abuse symptoms in high-risk adolescents. AB - Concerns about the cultural competence of child mental health services has led to the examination of racial/ethnic and gender differences in the prevalence of psychiatric symptoms. This study examines racial and gender differences in depressive and substance abuse symptomatology in a high-risk population of adolescents living in five residential group homes in South Carolina. We surveyed 299 youth ages 12 to 17, including 101 African American and 198 Whites. They completed the Centers for Epidemiological Studies-Depression scale (CES-D) and questions on substance abuse, demographics, and psychosocial functioning. No significant differences were found in the percentages of Whites and African Americans scoring above 16+ and 23+ cutoff scores on the CES-D, but significant gender differences were identified. Neither race nor race by age group interactions were found to be significantly correlated in regression analyses with CES-D score nor multiple substance use, whereas gender (p < .001) and school performance were significantly correlated with CES-D score, and poverty was correlated with multiple substance use. Our results indicate that levels of depressive symptomatology as measured by the CES-D are much more sensitive to gender than to race in high-risk populations. Different gender cutoffs are indicated when using systematic instruments in the measurement of depressive symptoms. PMID- 9225568 TI - Predictors of English fluency among Hmong refugees in Minnesota: a longitudinal study. AB - The objective of this study was to assess factors associated with later acquisition of English language fluency among Hmong refugees in Minnesota. Fluency in a society's lingua franca is a critical skill in psychosocial adaptation and mental health. A longitudinal study design was used, in which premigration and early postmigration factors were related to subsequent English fluency. The first group of 102 Hmong refugees located in Minnesota by the Immigration and Naturalization Service participated, and were interviewed in their homes. Hmong research assistants collected data using a questionnaire format at 1.5 years following resettlement in the U.S. Eight years later, two measures of English language competence were obtained: a self-assessment and an objective measure of English language fluency. Self-assessed fluency and performance on a brief English test showed good correlation. Greater English fluency on both measures was predicted by the following: younger age, male gender, education or vocational training in Laos prior to migration, occupation in Laos requiring literacy, study of English while in Asia, less proximity to other Hmong households in the U.S., any educational involvement in the U.S. (except English as a second language or ESL training), and not receiving welfare. Self-assessment of English fluency appeared to be a valid measure of competence in English. Demographic characteristics, certain premigration experiences, and early postmigration experiences predicted English fluency after 10 years in the U.S. ESL training was not associated with eventual English fluency on either self assessment or objective testing. Recommendations are made to enhance English fluency, and hence the psychosocial adaptation of refuguees and other immigrants to the U.S. PMID- 9225569 TI - Politics and South African mental health: hope, truth, and reconciliation. PMID- 9225570 TI - [Red cell Na+/H+ exchange and role of protein kinase C in its stimulation in diabetes mellitus, essential hypertension and nephropathy]. AB - Na+/H+ exchange (NHE) was measured as maximal initial velocity of pH-dependent H+ efflux from red cells into an alkaline medium containing Na+ in patients with insulin-dependent or noninsulin-dependent diabetes, with and without hypertension and in normoglycemic, essential hypertensives and normal controls (50 subjects in each subgroup). Maximal velocities of NHE were found in microalbuminuric patients in all subgroups, and NHE correlated with the rate of micro-albuminuria (r = 0.61, p = 0.02). Daily insulin requirements were greater in those with elevated NHE (84 +/- 8 vs 42 +/- 4 U/day). There was no correlation between NHE and levels of plasma glucose, HbA1 and plasma aldosterone and lipid profile and PRA. NHE was correlated with plasma prolactin (r = 0.51, p = 0.02) and PTH r = 0.24, p = 0.05). In uremic patients, NHE was inversely correlated with creatinine clearance (r = -0.48, p = 0.03). Since calphostin C, a selective inhibitor of protein kinase C, lowered increased NHE in vitro, the protein kinase C-dependent pathway of the exchanger regulation was concluded to be responsible for NHE activation in diabetes mellitus and essential hypertension. PMID- 9225571 TI - [Seminiferous tubule cytological pattern in infertile, azoospermic men in diagnosis and therapy]. AB - We determined spermatogenic patterns of seminiferous tubules in azoospermic infertile men and evaluated the prevalence of bilateral testicular homogeneity. 185 azoospermic men underwent bilateral testicular fine-needle aspiration (TFNA) in which each testis was punctured at 3 different positions. Aspirated material was stained and classified according to the most mature spermatogenic cell type present or whether only Sertoli cells were present. 35.7% had spermatozoa in their testes, 36.2% had spermatogenic maturation arrest, and 28.1% had only Sertoli cells in their seminiferous tubules. In 15.6% of all patients, the diagnosis in 1 testis differed from that in the other. In only 73.2% of those with testicular spermatozoa was it bilateral. In the remaining 26.9%, only Sertoli cells, spermatocytes or spermatids were found as the most mature cell type in the other testis. The study definitely indicates that fertilization with retrieved testicular spermatozoa should not be offered to azoospermic patients without prior evaluation of the seminiferous tubuespermatogenic pattern in both testes. PMID- 9225572 TI - [Attention deficit hyperactivity disorder, facilitating alcohol and drug abuse in an adult]. AB - Attention-deficit hyperactivity disorder (ADHD) has been considered a mental and behavioral disorder of childhood and adolescence. It is being increasingly recognized in adults, who may have psychiatric co-morbidity with secondary depression, or a tendency to drug and alcohol abuse. We describe a 32-year-old woman known for years as suffering from borderline personality disorder and drug dependence (including hashish, marijuana, LSD and "ecstasy") and alcohol abuse that did not respond to treatment. Only when correctly diagnosed as ADHD and appropriately treated with the psychotropic stimulant, methylphenidate (Ritalin), was there significant improvement. She succeeded academically, which had not been possible previously, the craving for drugs diminished and a drug-free state was reached. Although administration of psychostimulants to drug abusers is controversial, as they are addictive, in cases of ADHD they have promoted drug abstinence. PMID- 9225573 TI - [Toxic shock syndrome]. AB - Toxic shock syndrome (TSS) is a rare, life-threatening, acute multisystem illness usually characterized by sudden onset of high fever, diffuse sunburn-like erythroderma and a variety of other signs and symptoms. It may progress rapidly to hypotension and shock with multiple organ failure. Its exact cause is unknown, but in almost all cases there has been an infection with exotoxin-producing strains of phage group I Staphylococcus aureus. Although initially described in association with the use of superabsorbent tampons in menstruation, TSS has complicated a variety of surgical procedures. Recently in head and neck surgery attention has focused on absorbent packing materials, such as those used in postoperative nasal care. TSS developed in a 12-year-old 28 hours after tonsillectomy, nasal septoplasty and inferior turbinectomy in which absorbent packing material was used. It is important to maintain a high index of suspicion for TSS in all postoperative patients with fever, hypotension and erythroderma. PMID- 9225574 TI - [Angioedema caused by splenectomy with malignant lymphoma followed by multiple myeloma 7 years later]. AB - Acquired C1-inhibitor (C1-INH) deficiency has been reported in patients with immunoglobulin abnormalities and lymphoproliferative disorders, and angioedema has appeared simultaneously with the lymphoproliferative disease. We present a 50 year-old woman with acquired C1-INH deficiency and angioedema which preceded by 7 years the diagnosis of malignant mantle cell lymphoma. During the interval she was treated with Danazole and there were no attacks of angioedema. When routine follow-up bone marrow aspiration revealed infiltration of nonspecified lymphoma cells, exploratory laparotomy and splenectomy were performed. A month later Danazol was stopped, C1-INH levels returned to normal and there were no attacks of angioedema. Mantle cell lymphoma consisting of lymphocytes with cytoplasmic IgM-lambda was diagnosed in the excised spleen but chemotherapy was not initiated. 6 months later, a second lymphoproliferative disorder, multiple myeloma IgA kappa, was diagnosed. PMID- 9225575 TI - [Recurrent chronic multifocal bone infection]. AB - We present a 9-year-old girl who had chronic recurrent multifocal osteomyelitis. The bones involved were: right clavicle, distal fibula (bilateral), left sacroiliac and right wrist. After 10 years of follow-up; she is asymptomatic but presents radiological evidence of lesions in the right clavicle and left sacroiliac joint. The diagnosis was made by exclusion criteria. The biopsy and results of cultures from various bones were negative 4 times. Although chronic recurrent multifocal osteomyelitis is rare, it should be considered in the differential diagnosis of acute or chronic osteomyelitis and neoplasma. Its recognition avoids unnecessary laboratory tests and antibiotic therapy. PMID- 9225576 TI - [Gastrografin for mechanical partial, small bowel obstruction due to adhesions]. AB - The therapeutic effect of gastrografin is occasionally mentioned in the literature. However, this effect has not been objectively evaluated. We studied prospectively the effect of Gastrografin in cases of adhesive, simple, partial, small bowel obstruction (SBO) compared to conventional management. During 3 years, a total of 137 episodes of simple, partial SBO in 127 patients (10 recurrent episodes) were treated. The episodes were randomized into a control group (80 episodes), treated conventionally, and a trial group (77 episodes), which received in addition 100 ml of Gastrografin administered through the nasogastric tube. The two groups were well-matched with regard to age, gender, weight, medical and surgical background and duration of complaints before admission. Time to first stool and resolution of obstruction, complications, need for surgery, and hospital stay were noted. Mean time to first stool was significantly shorter in the trial group: 6.2 +/- 3.9 hours vs 23.5 +/- 12.7 (p < .0001). Mean hospital stay for unoperated patients was also shorter in the trial group: 2.7 +/- 2 days vs 5.5 +/- 2 days, (p < .0001). In addition, significantly fewer episodes in the trial group required operation, 10.4 vs 26.7% (p < 0.013). 1 patient in each group dies following operation. There were no Gastrografin related complications and it was effective and safe for adhesive, partial, simple SBO. It significantly speeds resolution of obstruction, reduces the need for operation, and shortens convalescence. PMID- 9225577 TI - [Gastric duplication cyst in an adult]. AB - Gastric duplication cysts are rare in adults and usually asymptomatic. In most cases they are discovered incidentally by abdominal ultrasound, CT, or upper gastrointestinal x-rays. Most of the duplications (82%) are cystic and do not communicate with the stomach. Approximately half of the cases are associated with other congenital anomalies. We report a 59-year-old woman operated on for a pancreatic mass that proved to be a gastric duplication cyst. The cyst was resected and the postoperative course was uneventful. PMID- 9225578 TI - [Systemic lupus erythematosus in Israel]. PMID- 9225579 TI - [Autoimmune phenomena and musculoskeletal manifestations in hepatitis C viral infection]. PMID- 9225580 TI - [Hepatitis C, therapy and predicting factors]. PMID- 9225581 TI - [Prenatal diagnosis of orthopedic anomalies]. PMID- 9225582 TI - [The importance of acute pain units]. PMID- 9225583 TI - [Developmental dysplasia of the hip]. PMID- 9225584 TI - [Short bowel syndrome]. PMID- 9225585 TI - [Pathogenesis of hemolytic uremic syndrome and thrombotic thrombocytopenia purpura]. PMID- 9225586 TI - [A new generation of infections in a changing world. Infectious diseases at the end of the twentieth century]. PMID- 9225588 TI - [Regional health care administration following the Netaniahu Commission]. PMID- 9225587 TI - [Methylphenidate (Ritalin) treatment in attention deficit hyperactivity disorder]. PMID- 9225589 TI - Ischemic stroke syndromes in childhood. AB - The ischemic stroke syndrome is very broad and encompasses a wide range of underlying conditions. Its identification is of great importance in clinical routine, in particular in the management of young patients who have acute neurologic deficits. The introduction of CT, MR and ultra-sound demonstrating lesions of the brain and in certain degree of the cerebral arteries has in general eliminated the need for angiography as a first examination. The most common underlying anomaly found with thrombotic or embolic stroke is congenital or acquired heart disease. Thus, it is essential that patients with cerebral ischemia be submitted to a complete cardiac examination. Children tend to show more recovery after a stroke than adults do. PMID- 9225590 TI - Esophageal ulcers in AIDS. AB - Thirty five esophageal biopsies from AIDS patients with clinical symptoms of esophagitis sent to "Emilio Ribas Institute", Pathology Laboratory, in a 2 year period were revised for possible infectious agents. Microorganisms were seen in 17 cases (48.6%). In 6 cases (17.1%), Acid-Fast bacilli were observed. One of these cases also had characteristic cytomegalic inclusions in endothelial cells. Inflammatory responses were composed of lymphocytes, some plasma cells and many histiocytes, with absence of giant cells in 4 cases of mycobacteriosis; in the other 2 cases, acid-fast bacilli were seen over the epithelium. Exclusive infection by cytomegalovirus was detected in 5 cases (14.3%), and candidiasis in 5 cases (14.3%). In one case there was association of cytomegalovirus and candidiasis. Esophageal ulcers in AIDS patients caused by Mycobacterium sp, may be more common than previously reported, and certainly an overlooked diagnosis. Once esophageal biopsy is an easy diagnostic procedure, this method may be used in routine screening for tuberculosis in patients with AIDS. PMID- 9225591 TI - Chromosome localization of the gene for growth hormone in the common shrew (Sorex araneus). AB - The chromosome localization of the gene encoding growth hormone (GH) was determined by Southern blotting of DNA obtained from a panel of common shrew x Chinese hamster and common shrew x mouse hybrid somatic cell clones using mink GH DNA as a probe. The GH gene was found to be localized on chromosome hn of the common shrew. PMID- 9225592 TI - Fucosylated glycoproteins in Chinese hamster metaphase chromosomes. AB - Distribution of fucosylated proteins in Chinese hamster metaphase chromosomes was studied with a fluorescein isothiocyanate conjugated monofucosyl-specific lectin, Ulex europaeus agglutinin I (UEA I). In situ binding of UEA I showed that fucosylated chromosomal proteins are preferentially localized to two internal structural domains of metaphase chromosomes: the helically coiled substructure of chromatids, and the Q-band regions. Corresponding proteins were identified in Western blots of isolated metaphase chromosomes. Several proteins with molecular weights ranging from 33 to 195 kD were recognized by UEA I. These data suggest that a subset of chromosomal proteins are fucosylated; they may have a role in the structural organization of chromosomes. PMID- 9225593 TI - Origin of an oral adenocarcinoma as suggested by cytogenetical observations. PMID- 9225595 TI - The association between arthritis and the weather. AB - Despite the prevasiveness of the idea that arthritis is influenced by the weather, scientific evidence on the matter is sparse and non-conclusive. This study, conducted in the Australian inland city of Bendigo, sought to establish a possible relationship between the pain and rigidity of arthritis and the weather variables of temperature, relative humidity, barometric pressure, wind speed and precipitation. Pain and rigidity levels were scored by 25 participants with osteoarthritis and/or rheumatoid arthritis four times per day for 1 month from each season. Mean pain and rigidity scores for each time of each day were found to be correlated with the meteorological data. Correlations between mean symptoms and temperature and relative humidity were significant (P < 0.001). Time of day was included in the analysis. Stepwise multiple regression analysis indicated that meteorological variables and time of day accounted for 38% of the variance in mean pain and 20% of the variance in mean rigidity when data of all months were considered. A post-study telephone questionnaire indicated 92% of participants perceived their symptoms to be influenced by the weather, while 48% claimed to be able to predict the weather according to their symptoms. Hence, the results suggest (1) decreased temperature is associated with both increased pain and increased rigidity and (2) increased relative humidity is associated with increased pain and rigidity in arthritis sufferers. PMID- 9225594 TI - The time factor in mortality: weather associations in a subtropical environment. AB - Mortality rates for a decade in Brisbane are analysed for dependence upon atmospheric factors. Time filters are applied to both the dependent and independent variables, and several models are developed to enable prediction, especially for weekly intervals. Statistically, deaths are observed to increase with colder and less humid weather with winds from a westerly, direction. Overall, taking account of both synoptic and seasonal influences, > 90% of cumulative deviations from mean death rates are explained. Some differences are also noted in the association of death with the weather between sexes, age groups and causes of death. PMID- 9225596 TI - Effects of acute hyperthermia on the carotid baroreflex control of heart rate in humans. AB - The purpose of this study was to examine the effect of hyperthermia on the carotid baroreceptor-cardiac reflexes in humans. Nine healthy males underwent acute hyperthermia (esophageal temperature -38.0 degrees C) produced by hot water perfused suits. Beat-to-beat heart rate (HR) responses were determined during positive and negative R-were-triggered neck pressure steps from +40 to -65 mm Hg during normothermia and hyperthermia. The carotid baroreceptor-cardiac reflex sensitivity was evaluated from the maximum slope of the HR response to changes in carotid distending pressure. Buffering capacity of the HR response to carotid distending pressure was evaluated in % from a reference point calculated as (HR at 0 mm Hg neck pressure-minimum HR)/HR range x 100. An upward shift of the curve was evident in hyperthermia because HR increased from 57.7 +/- 2.4 beats/min in normothermia to 88.7 +/- 4.1 beats/min in hyperthermia (P < 0.05) without changes in mean arterial pressure. The maximum slope of the curve in hyperthermia was similar to that in normothermia. The reference point was increased (P < 0.05) during hyperthermia. These results suggest that the sensitivity of the carotid baroreflex of HR remains unchanged in hyperthermia. However, the capacity for tachycardia response to rapid onset of hypotension is reduced and the capacity for bradycardia response to sudden hypertension is increased during acute hyperthermia. PMID- 9225597 TI - Time series analysis supporting the hypothesis that enhanced cosmic radiation during germ cell formation can increase breast cancer mortality in germ cell cohorts. AB - Techniques from cancer epidemiology and time series analysis were used to explore the hypothesis that cosmic radiation can induce germ cell changes leading to increases in future breast cancer mortality. A birth cohort time series for female breast cancer mortality was obtained using a model-independent, age-period cohort analysis on age-specific mortality data for 1940-1990. The birth cohort series contained several oscillatory components, which were isolated and compared to the corresponding frequency components of a cosmic ray surrogate time series Greenland ice-core 10Be concentrations. A technique, referred to as component wave-train alignment, was used to show that the breast cancer and cosmic ray oscillations were phase-locked approx. 25 years before the time of birth. This is consistent with the time of germ cell formation, which occurs during the fetal development stage of the preceding generation. Evidence is presented that the observable oscillations in the birth cohort series were residues of oscillations of much larger amplitude in the germ cell cohort, which were attenuated by the effect of the broad maternal age distribution. It is predicted that a minimum of 50% of breast cancer risk is associated with germ cell damage by cosmic radiation (priming event), which leads to the development of individuals with a higher risk of breast cancer. It is proposed that the priming event, by preceding other steps of carcinogenesis, works in concert with risk factor exposure during life. The priming event is consistent with epigenetic changes such as imprinting. PMID- 9225598 TI - Antimicrobial activity of some 13-alkyl substituted protoberberinium salts. AB - Several 13-alkyl substituted analogs of berberine and palmatine were found to be highly active against two types of Staphylococcus aureus (S1 and S2) of different origin. The most active 13-hexylberberine was 8 times more active (against S1) and the same order active (against S2) as kanamycin sulfate. 13-Hexylpalmatine displayed an activity against S. aureus (S1) 4 times greater than that of kanamycin sulfate. The activities of 13-hexylberberine against two types of S. aureus were 64 and 128 times greater than those of the clinically used alkaloid berberine. Additionally two hexyl derivatives possessed antifungal activity. PMID- 9225599 TI - Anti-inflammatory activity of aqueous extracts and steroidal sapogenins of Agave americana. AB - Lyophilized aqueous extracts obtained from Agave americana L (Agavaceae) collected in the north of Sardinia were characterized with regard to their steroidal sapogenin content. Extracts of A. americana and genins isolated from them were evaluated for anti-inflammatory properties by testing their effects on carrageenin-induced edema. The effect of orally administered genins on gastric mucous membranes was also assessed. Lyophilized extracts administered by the intraperitoneal route at doses equivalent to 200 and 300 mg/kg of fresh plant starting material, showed good anti-inflammatory activity. Doses of genins (total steroidal sapogenins, hecogenin and tigogenin) equivalent to the amount in the lyophilized extracts produced an antiedentatous effect which was much stronger and more efficacious than that obtained with an i.p. administration of 5 mg/kg of indomethacin or dexamethasone 21-phosphate at a dose equivalent to the molar content of hecogenin administered. At the doses used to evaluate the anti inflammatory activity, the genins did not have any harmful effect on the gastric mucous membranes. Lesions occurred when significantly higher doses of hecogenin were given, but gastric damage was still less than that caused by the drugs used for comparative purposes. PMID- 9225600 TI - Anti-allergic properties of the natural PAF antagonist yangambin. AB - In this study we examined the ability of the furofuran lignan yangambin to influence the local and systemic responses induced by antigen or PAF in actively sensitized or normal rats. Given intraperitoneally 1 h before stimulation, yangambin inhibited the pleural neutrophil and eosinophil infiltration evoked by the i.pl. injection of PAF or antigen into normal or 14 daysensitized rats whereas plasma exudation evoked by both stimuli was unaffected. The pleural neutrophil influx (6 h) after LTB4 stimulation was also significantly inhibited by yangambin. We also evidenced that the hemoconcentration, thrombocytopenia, and leucocytosis noted after i.v. PAF were all attenuated by yangambin. In actively sensitized rats, pretreatment with yangambin failed to modify the antigen-induced hemoconcentration and leucocytosis, but dose-dependently abrogated the thrombocytopenia noted 1 h post-stimulation. In vitro, the anaphylactic contraction of longitudinal jejunal segments to antigen challenge was significantly inhibited by yangambin (10(-5)-10(-4) M). Likewise, the contraction of jejunal segments from normal rats to PAF was markedly blocked by yangambin under conditions where the response to 5-hydroxytryptamine (5-HT) was not altered. In conclusion, our results show that antigen- and PAF-induced pleural neutrophil and eosinophil accumulation, but not exudation, is sensitive to treatment with yangambin. In addition, yangambin also suppressed the pleural neutrophil infiltration triggered by LTB4 as well as the blood thrombocytopenia and intestinal anaphylaxis elicited by antigen in rats. Thus, our findings indicate that yangambin shows an antagonistic action on receptors other than those of PAF, i.e., LTB4, and strongly suggest that it may be a useful drug in the treatment of some allergic inflammatory responses. PMID- 9225601 TI - Antioxidative action of diterpenoids from Podocarpus nagi. AB - Diterpenoids, totarol (1), totaradiol (2), 19-hydroxytotarol (3), totaral (4), 4 beta-carboxy-19-nortotarol (5), sugiol (6), isolated from Podocarpus nagi, were evaluated as antioxidants. Microsomal lipid peroxidation induced by Fe(III) ADP/NADPH and mitochondrial lipid peroxidation induced by Fe(III)-ADP/ NADH were inhibited by these terpenoids. They inhibited linoleic acid autoxidation but not generation of superoxide anion. Totarol (1) protected mitochondrial respiratory enzyme activities against NADPH induced oxidative injury. Totarane diterpenes from P. nagi were shown to be effective to protect biological systems and function against various oxidative stresses. PMID- 9225603 TI - Antinociceptive components of Ganoderma lucidum. AB - The antinociceptive effects 134 extracts prepared from 45 species of mushrooms were examined by the acetic acid-induced writhing method. From the CH2Cl2 extract of Ganoderma lucidum among the active extracts, ganoderic acids A, B, G and H and compound C6 were isolated as the antinociceptive components. PMID- 9225604 TI - Bicuculline-induced epileptiform activity in rat hippocampal slices: suppression by Aconitum alkaloids. AB - Alkaloids of Aconitum spec. (Ranunculaceae) are employed in traditional Chinese folk medicine as analgesics. The present study was designed in order to investigate the effects of the structurally related alkaloids aconitine, lappaconitine, and 6-benzoylheteratisine on experimentally induced epileptiform activity. Experiments were performed as extracellular recordings of stimulus evoked population spikes in rat hippocampal slices. Epileptiform activity was induced by bicuculline. All three alkaloids exerted an inhibitory action on excitability of hippocampal pyramidal cells in a frequency-dependent manner. The onset of inhibition was accelerated by increasing the frequency of electrical stimulation. Aconitine (1 microM) evoked a complete suppression of both normal and epileptiform activity, whereas lappaconitine (10 microM) and 6 benzoylheteratisine (10 microM) selectively diminished the epileptiform afterdischarges and the duration of the bursts, but spared the normal activity. The present findings suggest that the structurally related Aconitum alkaloids aconitine, lappaconitine, and 6-benzoylheteratisine possess an anticonvulsive potential. The predominant effect of these alkaloids is to suppress the spread of seizure activity, and they may therefore tend to distort epileptic events. However, despite their similar structure, they exert qualitatively and quantitatively different inhibitory effects. PMID- 9225602 TI - Immunostimulating activity of Celosian, an antihepatotoxic polysaccharide isolated from Celosia argentea. AB - Celosian, an acidic polysaccharide from the seeds of Celosia argenteo (Amaranthaceae) was found to be a potent antihepatotoxic agent for chemical and immunological liver injury models in animals. The immunomodulating action of celosian was studied to clarify the preventive mechanism of celosian on liver injuries. Celosian induced tumor necrosis factor-alpha (TNF-alpha) production in mice. Celosian also induced the production of interleukin-1 beta (IL-1 beta) and nitric oxide (NO) in macrophage cell line J774.1 in a concentration-dependent manner (1 to 1000 micrograms/ml). Moreover, celosian induced IL-1 beta secretion in human mononuclear cells. In addition, celosian enhanced gamma interferon (IFN gamma) production activity of concanavalin A (Con A) in mice spleen cells, though celosian alone did not significantly influence IFN-gamma production. These results indicate that celosian is an immunostimulating agent in addition to antihepatotoxic effects. PMID- 9225606 TI - The influence of Vicia faba (broad bean) seedlings on urinary sodium excretion. AB - Dopamine (DA) is known to increase diuresis and natriuresis through its action on renal dopaminergic receptors. Augmentation of intra-renal DA concentration by enhancement of its in situ production is greatly dependent on the availability of its precursor L-DOPA to the sites of its renal decarboxylation. Vicia faba (Vf) is a ubiquitous plant rich in easily absorbable L-DOPA. Following ingestion of 40 g freshly chopped Vf containing 120-130 mg of L-DOPA, plasma L-DOPA and urinary sodium and DA excretion increased significantly. The DA/Cre ratio reached a maximum level (280 +/- 58 micrograms/g) 60 minutes after Vf ingestion. This was significantly higher than the DA/Cre ratio after a control meal (1.8 +/- 0.2 micrograms/g; P < 0.0005). The Na/Cre ratio reached the maximal level (2.85 +/- 0.42 mmol/g) 90 minutes after Vf ingestion. This was significantly higher than the Na/ Cre ratio after the control meal (1.4 +/- 0.24 mmol/g; P < 0.005). We suggest that Vf might be of value in treating conditions such as hypertension, heart failure, renal failure, and liver cirrhosis in which natriuresis and diuresis are medically beneficial. PMID- 9225605 TI - Effects of visnadine on rat isolated vascular smooth muscles. AB - Visnadine, an active principle extracted from the fruit of Ammi visnaga, exhibits peripheral and coronary vasodilator activities and has been used for the treatment of angina pectoris. The present study was undertaken to further characterize the inhibitory effects of visnadine on the contractile responses in rat isolated aortic rings and portal vein segments. Visnadine (< 10(-5) M) selectively inhibited the contractions induced by depolarization with 80 mM KCl or by CaCl2 in KCl-depolarized aorta and the spontaneous activity of the portal vein. Its inhibitory effects were not increased as the time of depolarization was prolonged and were similar in aorta incubated in 5 or 40 mM KCl. At concentrations higher than 10(-5) M, visnadine also inhibited the contractile responses induced by noradrenaline and phorbol 12-myristate 13-acetate (PMA), being equipotent to inhibit noradrenaline-induced contractions in either Ca(2+) containing or Ca(2+)-free medium and PMA-induced contractions. In conclusion, the present results suggest that visnadine preferentially inhibited the contractile responses mediated by Ca2+ entry through L-type Ca2+ channels, whereas at high concentrations it may also interfere with other sites involved in vascular smooth muscle contraction. PMID- 9225607 TI - Minor components with smooth muscle relaxing properties from scented myrrh (Commiphora guidotti). AB - All sesquiterpenes present in a sample of scented myrrh were isolated and characterised. Seven compounds, with cadinane, guaiane, oplopane, and eudesmane skeletons, were obtained, of which two are new and two are reported from a natural source for the first time. The major component, T-cadinol, has previously been shown to possess smooth muscle-relaxing properties, and the major purpose of the investigation was to compare the effects of the minor and more polar sesquiterpenes with that of T-cadinol in the rat aorta. Like T-cadinol, the minor sesquiterpenes are more efficient in reducing K(+)-induced contractions than those induced by the alpha-adrenoceptor agonist phenylephrine, however, they were all less potent than T-cadinol. PMID- 9225608 TI - Betulinic acid: isolation from Triphyophyllum peltatum and Ancistrocladus heyneanus, antimalarial activity, and crystal structure of the benzyl ester. AB - The known lupane-type triterpene betulinic acid (3) was isolated for the first time from Triphyophyllum peltatum and Ancistrocladus heyneanus. It was found to exhibit moderate to good in vitro antimalarial activity against asexual erythrocytic stages of the human malaria parasite Plasmodium falciparum. A first X-ray structure analysis succeeded after conversion into its benzyl ester 4. PMID- 9225609 TI - Steroidal saponins from Asparagus officinalis and their cytotoxic activity. AB - Two oligofurostanosides were isolated from the seeds of Asparagus officinalis L and their structures characterized as 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-(alpha L-rhamnopyranosyl- (1-->4))-beta-D-glucopyranosyl]-26-O-[beta-D-glucopyranosyl] (25R) -22 alpha-methoxyfurost-5-ene-3 beta,26-diol(methyl protodioscin) and its corresponding 22 alpha-hydroxy analogue (protodioscin). The structural identification was performed using detailed analysis of 1H- and 13C-NMR spectra including two-dimensional NMR spectroscopy (COSY, HMQC, NOESY and HMBC), and chemical conversions. These two compounds have been shown to inhibit the growth of human leukemia HL-60 cells in culture and macromolecular synthesis in a dose dependent manner. The inhibitory effect on DNA synthesis was found to be irreversible. PMID- 9225610 TI - Aged garlic extract and its constituents inhibit Cu(2+)-induced oxidative modification of low density lipoprotein. AB - Oxygen radical injury and lipid peroxidation have been suggested as major causes of atherosclerosis, cancer, liver disease, and the aging process. More specifically, oxidative modification of low density lipoprotein (LDL) has been recognized as an important process of atherosclerosis. In this study, we determined the effects of aged garlic extract (AGE), four of its constituents, and a metabolite on Cu(2+)-induced oxidative modification of LDL using an in vitro system. All these compounds were shown to inhibit oxidative modification of LDL. PMID- 9225611 TI - Protective effect of curcuminoids on epidermal skin cells under free oxygen radical stress. AB - Curcuminoids from Curcuma longa L. (Zingiberaceae) protected normal human keratinocytes from hypoxanthine/ xanthine oxidase injury. Since curcuminoids synergistically inhibited nitroblue tetrazolium reduction, a decrease in superoxide radical formation leading to lower levels of cytotoxic hydrogen peroxide was proposed as an explanation for this protective effect. PMID- 9225612 TI - Inhibition of phospholipase C gamma 1 by the prenylated flavonoids from Sophora flavescens. AB - The effect on the phospholipase C gamma 1 activity of eleven prenylated flavonoids from Sophora flavescens was investigated. These flavonoids exhibited relatively strong inhibitory activity with IC50 values ranged from 7.5 x 10(-6) M to 35 x 10(-5) M with the exception of kushenol H (4) (IC50 value; > 5.3 x 10(-4) M). The presence of C3-OH resulted in a significant diminution of activity and the configuration of C3-OH is likely to be another factor influencing the activity. In addition, hydration of the C-4"'-C-5"' double bond of the lavandulyl side chain caused complete loss of activity. These data suggest that the presence and configuration of C3-OH are related to the inhibitory activity and the lavandulyl side chain is also important for high inhibitory activity against PLC gamma 1. PMID- 9225613 TI - Phenolic and antibacterial constituents of Vahlia capensis. AB - The n-butanol fraction of Vahlia capensis yielded kaempferol, quercetin, afzelin, astragalin, quercitrin, isoquercitrin, rutin, gallic acid, chiro-inositol, dulcitol, and a novel biflavonoid, VC-15B (vahlia biflavone). The compounds were identified using 1D and 2D NMR techniques and FABMS. Vahlia biflavone and gallic acid were isolated, using bioassay-guided procedure and identified as the antibacterial components. Both compounds showed activity against Gram positive Staphylococcus aureus and Bacillus subtilis. Vahlia biflavone gave MIC values of 15.3 micrograms/ml and 30.6 micrograms/ml against S. aureus and B. subtilis, respectively while gallic acid gave a value of 71.3 micrograms/ml for both organisms. PMID- 9225614 TI - Composition and antimalarial activity in vitro of the essential oil of Tetradenia riparia. AB - The essential oil from the leaves and stems of Tetradenia riparia was analysed by GC and GC/MS and 35 components were identified. The main constituents were alpha terpineol (22.6%), fenchone (13.6%), beta-fenchyl alcohol (10.7%), beta caryophyllene (7.9%), and perillyl alcohol (6.0%). Moderate antimalarial activities were recorded against two strains of Plasmodium falciparum. PMID- 9225615 TI - Anti-tumor promoting activities of lantadenes on mouse skin tumors and mouse hepatic tumors. AB - Two-stage carcinogenesis of mouse skin papillomas induced by 7,12 dimethylbenz[alpha]anthracene and 12-O-tetradecanoylphorbol 13-acetate, and mouse hepatic tumors induced by N-nitrosodiethylamine and phenobarbital, were inhibited by lantadenes. PMID- 9225616 TI - Bicycloillicinone asarone acetal: a novel prenylated C6-C3 compound increasing choline acetyltransferase (ChAT) activity from Illicium tashiroi. AB - Bicycloillicinone asarone acetal (1), a novel bicyclic prenylated C6-C3 compound, has been isolated from the woods of Illicium tashiroi and its structure has been elucidated by spectroscopic analyses and chemical degradation. Compound 1 was found to increase choline acetyltransferase (ChAT) activity in culture of P10 rat septal neurons. PMID- 9225617 TI - Effects of aging on implicit sequence learning: accounting for sequence structure and explicit knowledge. AB - The present research was intended to examine the sequence learning ability of elderly people-with a focus on comparing sequences with different structural characteristics and on properly assessing explicit knowledge. Experiment 1 showed that learning-related improvements in serial reaction time task performance were greater for young than elderly subjects, and elderly subjects were especially poor at learning a sequence with complex structural characteristics. Measures of recognition memory showed that neither young nor elderly subjects showed above chance explicit knowledge of the sequences. Experiment 2 was designed to test the validity and sensitivity of the explicit recognition measures by comparing young subjects in groups given all random trials, given sequence trials with implicit instructions, or given sequence trials with explicit instructions. Experiment 2 confirmed the sensitivity of the recognition measures to explicit knowledge, so it is concluded that group effects in Exp. 1 reflect age-related differences in implicit learning. PMID- 9225618 TI - Learning spatial sequences in unilateral neglect. AB - Brain-damaged patients with unilateral spatial neglect ignore aspects of the world located on the side opposite their lesion. In the present study we examined the performance of unilateral neglect patients (UN) on an SRT task in which a hybrid repeating sequence (21313) was used. We analyzed the patients' performance for each location separately as a function of the target's location in the trial preceding the response. The UN patients were severely limited in their learning of the sequence when compared to normal controls. In particular, they appeared to learn unique associations (21 and 13) but not ambiguous ones (31 and 32). We discuss two possible explanations for this phenomenon. The first is that UN patients show a deficit similar to that of normal subjects in dual task situations. The second is that the learning deficit is unique to spatial processing impairments of UN patients and is not directly related to research with normal population. We outline future research that may distinguish between these two explanations. PMID- 9225619 TI - Effects of rate of reinforcement and rate of change on choice behaviour in transition. AB - In two experiments with pigeons, a single variable-interval schedule assigned reinforcers to two response keys on a percentage basis. The percentage of reinforcers assigned to each key was changed every few sessions, and subjects' choice responses were recorded before and after each change. In Experiment 1, the overall rate of reinforcement was varied across conditions. The pigeons' choice responses adapted more quickly to a change in the reinforcement percentages when the overall reinforcement rates were higher, but acquisition rates varied by only about a factor of 3, whereas reinforcement rates were varied by about a factor of 9. In Experiment 2, the reinforcement percentages changed about every 8 sessions in Phases 1 and 3, but every 1 or 2 sessions in Phase 2. Pigeons' choice responses adapted to a change in reinforcement percentages more quickly in Phase 2 than in Phases 1 and 3. The results from both experiments pose difficulties for several prominent models of transitional choice behaviour. The results suggest that each successive reinforcer has more impact on a subject's subsequent choice behaviour when the overall rate of reinforcement is lower and when the reinforcement contingencies have changed frequently in the recent past. PMID- 9225620 TI - Weight loss in rats produced by running: effects of prior experience and individual housing. AB - To investigate factors affecting activity-based anorexia (ABA) or activity-stress (AS), rats were given 2-hr access to a running wheel immediately prior to their daily 1.5-hr food access during the light cycle. This produced a reduction in food intake, a steady increase in running, and a large drop in body weight with a prolonged delay before weight recovery began. Experiment 1 found that these effects were reduced in rats with prior experience of eating at this time of day. In contrast, prior experience of running in the wheel when on ad lib food enhanced these effects in Experiment 2, where a subsequent change for half the subjects to individual housing produced a further decrease in body weight. The latter factor was investigated from the outset of Experiment 3 and again individually housed rats showed greater weight loss than did group-housed rats. This experiment also found that in rats of the same age a low initial body weight predicts greater vulnerability to ABA. It was concluded that ABA results from activity-induced reduction of feeding, which prolongs adaptation to a new feeding schedule and is accentuated by social isolation. PMID- 9225621 TI - The effects of hippocampal and area parahippocampalis lesions in pigeons: I. Delayed matching to sample. AB - Four experiments were conducted to determine the effects of bilateral damage to the hippocampus and area parahippocampalis (Hp-APH) on visual memory in pigeons using the delayed matching-to-sample (DMS) procedure. In Experiment 1, we generated visual retention gradients with delays of 0, 1.5, 3, 6, and 12 sec both preoperatively and postoperatively in three pigeons with considerable preoperative visual DMS experience. Bilateral Hp-APH lesions had no effect whatsoever on visual retention. In Experiment 2, we examined the effects of Hp APH lesions on both the acquisition of a visual DMS task with a 0-sec delay, and the subsequent retention performance with delays of 0, 3, 6, 12, and 24 sec. There was no difference between unoperated control pigeons and Hp-APH pigeons either in terms of the number of sessions required to learn the visual DMS task or in terms of their subsequent visual retention performance levels. In Experiments 3 and 4, we examined whether Hp-APH pigeons might be more sensitive than control pigeons to the effects of proactive interference (by reducing the duration of the intertrial interval) and retroactive interference (by introducing delay-interval illumination). Although reducing the duration of the intertrial interval and increasing the level of delay-interval illumination both resulted in lower performance levels on the visual DMS task, there was no indication that the Hp-APH pigeons were any more affected by the changes in interference levels than were unoperated control pigeons. These findings support the view that the Hp-APH in pigeons plays little role in the processing and retention of purely visual information. PMID- 9225622 TI - The effects of hippocampal and area parahippocampalis lesions in pigeons: II. Concurrent discrimination and spatial memory. AB - Two experiments were conducted to examine the effects of bilateral hippocampus (Hp) and area parahippocampalis (APH) lesions in pigeons on the acquisition of a visual and spatial task. In Experiment 1, pigeons were trained on three successive six-pair concurrent discrimination tasks, each using a novel set of stimuli. There was no difference between control unoperated pigeons and Hp-APH pigeons in terms of the number of sessions required to learn either the first, second, or third concurrent discrimination task. In Experiment 2, the same pigeons were trained on an open-field spatial task similar in many ways to the radial-arm maze task used with rats. In contrast to the absence of impairments on the visual concurrent discrimination task, pigeons with Hp-APH lesions were severely impaired on the acquisition of the spatial task. These findings support the view that the Hp-APH in pigeons is important for the processing of spatial, rather than visual information. PMID- 9225623 TI - Categorization and perceptual learning: an analogue of the face inversion effect. AB - This paper reports two experiments that investigate the extent to which it is plausible to suppose that an associatively based mechanism for perceptual learning acts as the basis for the effects of inversion on identification, recognition, matching and discrimination of faces (and certain other stimuli rendered familiar by expertise, e.g. gundogs). In the first experiment, an inversion effect that is contingent both on familiarity with a category and on the category possessing prototypical structure is demonstrated using discrimination learning of chequerboard stimuli. The second experiment demonstrates that the inversion effect found in Experiment 1 can generalize to a recognition paradigm as well. These results are discussed within the framework provided by associative learning theory, and a parallel is drawn with models employing a norm-based coding in similarity space. The conclusion is that it would be remarkable if the inversion effects demonstrated with the abstract categories used in the experiments reported here were not implicated in the inversion effects found with other classes of stimuli, whilst conceding that the analogy is not complete, particularly in the case of faces. PMID- 9225624 TI - Investigating the relation between imagery and perception: evidence from face priming. AB - The relation between imagery and perception was investigated in face priming. Two experiments are reported in which subjects either saw or imagined the faces of celebrities. They were later given a speeded perceptual test (familiarity judgement to pictures of celebrities) or a speeded imagery test (in which they were told the names of celebrities and asked to make a decision about their appearance). Seeing faces primed the perceptual test, and imaging faces primed the imagery test; however, there was no priming between seeing and imaging faces. These results show that perception and imagery can be dissociated in normal subjects. In two further experiments, we examined the effects of imaging faces on a subsequent face-naming task and on a task requiring familiarity judgements to partial faces. Both these tasks were facilitated by prior imaging of faces. These results are discussed in relation to those of McDermott & Roediger (1994), who found that imagery promoted object priming in a perceptual test involving naming partial line drawings. The implications for models of face recognition are also discussed. PMID- 9225625 TI - Mood-state-dependent retrieval: the effects of induced mood on memory reconsidered. AB - Analysis of studies investigating mood-state-dependent retrieval identifies methodological problems that may have contributed to the controversy surrounding the reliability of the effect-in particular, the possible confounding of encoding and retrieval in previous studies. Five experiments are reported investigating the effects of mood on learning and recall. Mood-state-dependent retrieval was observed in Experiment 1a (using Velten's Mood Induction Procedure); Experiment 1b (using a music MIP); and Experiment lc (using Velten's MIP at encoding and a music MIP at retrieval). Subjects who learned and recalled in different moods had significantly greater decrements in recall than did subjects in the same moods. Experiments 2 and 3 investigated the effect of observable retrieval cues on mood state-dependent retrieval. In Experiment 2, the presence of observable retrieval cues at recall overrode state-dependent retrieval. In Experiment 3, by manipulating the presence or absence of observable cues at recall, both the occurrence and the erasure of the mood-state dependency was demonstrated. Mood state during learning and cued recall was also shown to affect performance in a third session under conditions of free recall. PMID- 9225626 TI - Disruption of short-term recognition memory for tones: streaming or interference? AB - A sequence of auditory stimuli interpolated between the initial presentation of a tone and a comparison tone impairs recognition performance. Notably, the impairment is much less with interpolated speech than with tones. Six experiments converge on the conclusion that this pattern of impairment is due more to the organization of the interpolated sequence than to its similarity to the to-be remembered standard. Factors that contribute to the coherence of the interpolated sequence into a stream distinct from the initial tone are primary determinants of the level of impairment. This is demonstrated by manipulating factors that contribute to the coherence of the interpolated sequence by the action of temporal, spatial, timbral, and tonal attributes. However, the relative immunity of recognition performance to the interpolation of unprocessed digit sequences is not explained wholly by such coherence. PMID- 9225629 TI - Clinical aspects on the use of preimplantation genetic diagnosis in couples at risk. AB - BACKGROUND: Preimplantation genetic diagnosis (PGD) is a new technique which may become an attractive alternative to traditional prenatal diagnosis for couples at risk of getting children with severe genetic diseases. We here report our experience after the first trials in Sweden. METHODS: On day three after fertilization by intracytoplasmic sperm injection (ICSI) one or two blastomeres were biopsied and diagnosed by fluorescence in situ hybridization (FISH). Gender determination was done in two cases where the female in the couples were carrier of a severe X-chromosome bound disease (Wiskott-Aldrich disease or ornithine transcarbamylase deficiency). RESULTS: The first couple got an embryo transfer in each of their two treatment cycles. Two female embryos were transferred in the first cycle and one in the second cycle. The second couple did not get any embryo transfer in their two cycles. No clinical pregnancy occurred. CONCLUSION: With the rapidly improving knowledge about inherited disease and refinement of the IVF techniques, PGD will play an important clinical role in high risk groups within a decade for both mono- and polygenic disorders. Both technical as well as legal and ethical problems have however to be solved before these new techniques can be applied on large scale. PMID- 9225628 TI - Fertility, pregnancy and cancer. AB - Results of clinical and experimental research into various aspects of cancer and pregnancy are reported. These investigations have been proceeding for many years. 845 cases of pregnancy with concomitant malignant tumor at various localization are reported here. Diagnostic methods and results of treating cervical and breast cancer during pregnancy are described. Also discussed is the influence of pregnancy on the clinical course of cancer and survival. Fertility after recovery from cancer accomplished by means of organ-preserving surgery is commented on. PMID- 9225630 TI - Quality assurance--a process for improving perinatal care. AB - Quality assurance is an ongoing process, including not only the assessment and evaluation of the quality of care, but also the definition and implementation of strategies to improve the quality. The process of quality assurance must start from the identification of health problems and of the appropriate interventions to address those problems. In perinatal care, however, appropriate assessment of the effectiveness of diagnostic and therapeutic interventions and care routines has often been lacking. This symposium will discuss examples of the various steps in quality assurance in perinatal care from countries in the Baltic region. PMID- 9225631 TI - Recent developments in perinatal problems in the Baltic countries. PMID- 9225632 TI - Perinatal problems and quality assurance in Latvia--a country in economic transition. AB - PURPOSE: To characterize the perinatal situation in Latvia today and to elaborate a program for future perinatal care. METHOD: We have analyzed the causes and structure of perinatal mortality at the Riga Maternity Home (RMH), a tertiary hospital where one-fifth of all Latvia's babies are born, and compared the results with Latvia in general. RESULTS: Since 1991, perinatal mortality has varied between 17 and 23 per 1000 births annually at the ORMH and between 17 and 19 per 1000 births in Latvia as a whole. According to official statistics, perinatal hypoxia is the main cause of perinatal death in about half of the cases. There is a high percentage of antenatal (45%) and intrapartum (12%) deaths. CONCLUSIONS: We have elaborated a number of measures to develop antenatal care and improve the intranatal care for term and preterm infants. PMID- 9225633 TI - Quality assurance in antenatal care--experiences from a professional workshop group in Sweden. PMID- 9225634 TI - A regional perinatal database in southern Sweden--a basis for quality assurance in obstetrics and neonatology. AB - BACKGROUND: In order to ensure as few avoidable adverse outcomes of pregnancy as possible, it is necessary to continuously evaluate the quality of both obstetric and neonatal care. The eleven southernmost hospitals in Sweden have joined together in a project of developing a regional database, with special emphasis on rapid output of information in order to identify changing trends. METHODS: A regional computerized database has been developed, collecting variables and quality indicators agreed upon by all participants. Specific protocols have been designed for obstetric care, neonatal care and autopsy findings. All participating units transfer information on paper forms or via local computerized information systems. The regional database thus receives data on about 20,000 deliveries annually. RESULTS: Data collection started on September 1, 1994. The first results are due in March 1996, and thereafter on a regular basis every 3 months. Special methods for rapid analysis of raw data have been developed with the help of commercially available data analysis tools. CONCLUSIONS: It is possible to construct an information system with different computer platforms and different database tools at each participating facility, as long as the database systems are locally controllable. That a software is commercially available is no guarantee that data transfer to a central database is possible. Experience from participating sites also indicates that a specialized database is needed for registering obstetric data, as general computerized record-keeping systems are unable to cope with an event concerning more than one subject at a time. PMID- 9225635 TI - The Nordic/Baltic perinatal death classification. PMID- 9225636 TI - Induced abortion--a global health problem. AB - Every year around 500,000 women are estimated to die from pregnancy-related causes, the majority in the developing world and many as a consequence of unsafe abortion. Around 25 per cent of maternal deaths in Asia and 30-50 per cent of maternal deaths in Africa and Latin America occur as a result of induced abortion. Data on abortion related maternal morbidity is less reliable than mortality but suggests that for every maternal death 10-15 women suffer significant pregnancy-related morbidity, i.e. infertility, genito-urinary problems and/or chronic pain. Induced abortion occurs in practically every society in the world but only 40 per cent of the women in the world live in countries where abortion is legally free. A permissive legislation is an important prerequisite for medically safe and early abortion. Oppositely, with a restrictive law, abortion is difficult to obtain, costly and possibly unsafe, in particular to the least affluent women in the society. Induced abortion in a developed country with legal and easy access to services is a safe procedure with hardly any mortality and very low morbidity. The best strategy to reduce the number of unsafe abortions is prevention of unwanted pregnancy. The consequences of unsafe abortion on women's health need to be acknowledged by everybody in the society in order to improve abortion care. It is necessary to adjust legal and other barriers to medically safe abortion in order to follow the declaration at the UN conference on population in Cairo, 1994, which stated that abortion, wherever legal, should be safe. It is also necessary to introduce preventive measures where abortions are performed, i.e. good and easily accessible family planning services. PMID- 9225638 TI - Pregnancy termination--situation in Finland. AB - Finnish abortion numbers have been declining steadily since the initial increase following the liberalization of abortion legislation in 1970, and in 1994, only 7.9 abortions per 1000 women between 15 and 49 years of age were performed. This figure is the lowest in the Nordic countries. Legislation, methods and complications, abortion services and contraceptive practices in Finland are described. PMID- 9225637 TI - Abortion in the framework of family planning in Estonia. AB - OBJECTIVE: To analyze the legal status of abortion and trends in childbirth, abortions and contraceptive use in Estonia. DESIGN: National statistical data on induced abortions, spontaneous abortions, births and contraceptive use in Estonia. RESULTS: Even though induced abortion is legal in Estonia, the abortion rate has been declining but is still high and in 1994 was 53.8 per 1000 women of fertile age. Among young women under 20 years of age, abortions decreased slightly in the period 1992-94 (from 55.5 to 41.5 per 1000). The birth rate has been declining rapidly in recent years, resulting in a net population reduction. The use of modern contraceptives is increasing but is still low. CONCLUSION: Survey on abortion and on contraceptive sales and use are needed. To improve family planning and prevent unwanted pregnancies it is important to increase awareness among the entire population--and especially among young people--by information, education and communication. A high standard of counseling, including pre- and postabortion counseling and in the youth services, is essential. PMID- 9225639 TI - Induced abortions in Denmark. AB - BACKGROUND: A law on Induced Abortion on Request came into force in Denmark in 1973. During the first years the rate of abortion increased but since the early 1980s the rate has been rather constant. The paper reviews recent findings concerning induced abortion and discusses its role in controlling fertility. METHODS: Trends in induced abortion is described from routine statistics while information on the aborting women are taken both from a survey and from a register based study of fertility- and abortion-pattern among a cohort of women. RESULTS: Fertility trends in Denmark are characterized by an increasing age at first birth. Half of the aborters to day have no children before and 10% had given birth less than 18 months earlier. Among aborters a higher proportion than among parturients were still under education and a higher proportion were single with no steady partner. Half of the aborters became pregnant in spite of contraceptive use, indicating a need for better contraceptives. CONCLUSIONS: Induced abortion has become a generally accepted form of birth control in Denmark and the decision to terminate a pregnancy is influenced by many factors including the woman's conjugal--and educational situation. A strategy for prevention of induced abortion must take into consideration the social circumstances of women and for families with children. PMID- 9225640 TI - Abortions in Malmo--problems and prevention. AB - Since 1975, women living in Sweden have had after counseling, free access to legal abortion until the 18th week of pregnancy. During the past decade, the abortion number remained around 34-38,000 per year, but since 1989 it has been decreasing continuously. This means an abortion rate ranging from 19.8 to 21.5 per 1,000 women aged 15-44. The largest towns in Sweden have the highest abortion rates, Malmo leading with 26.5-30.4 per 1,000 women. PMID- 9225641 TI - Clinical experience of a combined oral contraceptive with very low dose ethinyl estradiol. AB - BACKGROUND: The risk of thromboembolic events related to the ethinyl estradiol (EE) dose in oral contraceptive (OC) pills has led to a further dose reduction. METHODS: An OC pill with 150 micrograms desogestrel combined with only 20 micrograms EE was compared with a pill containing the same dose of desogestrel but 30 micrograms of EE in a Scandinavian multicentre study with follow-up visits after 3, 6 and 12 months. RESULTS: In almost 5,000 cycles with each pill the numbers of pregnancies due to method failure with the lower and higher EE dose pills were 0 and 2, respectively. Irregular bleedings were slightly more common with the lower EE dose, but tended to decrease over the year of study. Other side effects were uncommon. Regarding metabolic effects, both pills tended to raise the plasma HDL level and the lower EE dose pill also reduced LDL. Free testosterone was reduced by two-thirds with both pills, showing beneficial effects on acne. CONCLUSIONS: It is concluded that both these pills are reliable and safe, but that many women would accept a slightly greater risk of irregular bleeding with the 20 micrograms EE dose pill in exchange for a reduction in potential risk related to the estrogenic component of OC pills. PMID- 9225642 TI - Oral contraception and cerebral thromboembolism. PMID- 9225643 TI - Selection and performance of the levonorgestrel-releasing intrauterine system. AB - BACKGROUND: The levonorgestrel-releasing intrauterine system (LNg IUS) is a hormonal contraceptive that is used in the uterine cavity. To determine whether the reasons for choosing LNg IUS vs. copperreleasing intrauterine devices (Cu IUDs) differ and whether their performances are comparable, we carried out a retrospective study in Finland during the first years of LNg IUS use. METHODS: Gynecological and contraceptive histories of 626 LNg IUS and 626 Cu IUD users and the performance of the device were reviewed from patient records. RESULTS: Women who accepted the LNg IUS were more likely than Cu IUD acceptors to have a history of menstrual bleeding of 6 days or more (44.4% vs. 28.4%), heavy bleeding (44.8% vs. 8.4%) and moderate or severe dysmenorrhea (15.9% vs. 7.5%). In both groups, 70% of the women had used Cu IUDs earlier. However, the LNg IUS acceptors had had more side effects during previous use of CU IUDs (58.2% vs. 28.8%). They also reported more side effects that resulted in discontinuation of a previous Cu IUD (39.4% vs. 10.1%). However, the 12-month life-table continuation rates of 80.6 (SE 1.9)% vs. 83.4 (SE 1.8)% were alike. Cu IUD users discontinued the current method more often because of problems of bleeding and unwanted pregnancy. Among those women who had previously discontinued a hormonal method because of hormonal side effects, there were no differences in the continuation rates of the two groups. CONCLUSIONS: LNg IUS can be successfully used by women who cannot use a CU IUD or who have experienced hormonal side effects with oral contraceptives. PMID- 9225644 TI - The possible use of antiprogestins for contraception. AB - BACKGROUND: A number of compounds, antiprogestins, e.g. mifepristone, onapristone and lilopristone, have been developed which compete with progesterone at the receptor level. One of these, mifepristone, is in combination with a prostaglandin analogue currently in use for termination of early pregnancy. The possibility to use these compounds for contraceptive purposes is presently under evaluation. METHODS: The possible contraceptive effect of antiprogestins has been evaluated in both clinical and experimental studies. RESULTS: Administration of antiprogestin during the follicular phase has an inhibitory effect on follicular development and ovulation, and on endometrial development and function if administered during the secretory phase of the menstrual cycle. A high dose of mifepristone, 200 mg, administered immediately following ovulation is highly effective in preventing implantation, most likely due to an effect on endometrial receptivity. It seems that the endometrium is more sensitive to antiprogestin than is the ovulatory process. Low weekly, 2.5 mg to 5 mg, and daily doses, 0.5 mg, of mifepristone did not inhibit ovulation, but a significant effect on endometrial development and especially endometrial function judged from measurement of the expression of a number of markers for endometrial receptivity could be demonstrated. CONCLUSION: The effect of mifepristone on the endometrium may be sufficient to prevent implantation, and if so, an oral contraceptive method could be developed which has no effect on ovarian function. PMID- 9225645 TI - Quality assurance in endoscopic surgery. AB - A number of operative procedures in gynecology are now being performed with the help of endoscopy, a development that was not based upon appropriate clinical trials. Quality assurance is today's challenge in gynecological endoscopic surgery. Although as a rule data from quality registers cannot be used for comparative purposes, they may guarantee a basic standard of documentation of surgical results. Quality assurance is a continuous process and a means to improve quality in general. Quality indicators are specific criteria that denote quality change and play a key role in quality programs. It is important to realize that quality in this sense may be viewed not only from the medical aspect but from a variety of perspectives including education, training and medical care. PMID- 9225646 TI - Laparoscopic surgery in ectopic pregnancy. AB - BACKGROUND: A randomized, prospective clinical trial was conducted to compare the efficacy of laparoscopic treatment versus conventional conservative abdominal surgery for tubal pregnancy. METHODS: Patients were stratified for age and risk determinants for ectopic pregnancy (EP). Forty-eight patients were treated by laparoscopy and 57 by laparotomy. Entry criteria were: size of the ectopic gestation < 4 cm, hemodynamic stability, accessibility for laparoscopic treatment and a trained laparoscopist on duty. RESULTS: There was no difference between the groups regarding gestational duration, size and location of the ectopic gestation, and the mean preoperative hCG values. The groups did differ with respect to total operation time (73 min in the laparoscopy group vs. 88 min in the laparotomy group), hospital stay (2.2 days vs. 5.4 days) and convalescence period (11 days vs. 24 days). The rates of elimination of hCG were similar in the two groups, and there was no statistical difference in the rate of second intervention. CONCLUSIONS: Patients treated by laparoscopy had a shorter hospital stay and a shorter convalescence than patients from the laparotomy group. PMID- 9225647 TI - A renaissance for vaginal hysterectomy. AB - BACKGROUND: The most often performed major operation in gynecology is hysterectomy. The very first vaginal hysterectomy (VH) was performed by Langenbeck in 1813 (1). In 1934, Heany reported a series of 565 vaginal hysterectomies performed for benign diseases, half of which were indicated by fibromyomas (2). Since the 1950s many articles have been published on VH and indications for operation have increased. VH deserves more attention and needs to be carried out more often for the time of the operation is shortened, the postoperative complications occur less and the postoperative stay in hospital is shortened as well. METHODS: VH has been done with the traditional method. RESULTS: Between November 1, 1991 and April 1, 1995 we performed 38 VH operations. The average operating time was 55 minutes, not a single patient needed blood transfusion, one bladder rupture occurred and the average postoperative hospitalisation was 7.1 days. CONCLUSION: VH is a safe operation with few intra- and postoperative complications, without notable blood loss. VH allows one to shorten the operating time and allows early postoperative discharge of some patients from hospital. PMID- 9225648 TI - Subtotal versus total laparoscopic hysterectomy. AB - Between 1993 and 1994, 368 women underwent hysterectomy for benign disorders at the University of Kiel. Of these operations, 58.7% were performed either by pelviscopic or by laparotomic Classic Intrafascial Supracervical Hysterectomy, subtotal hysterectomy with coring of the inner cervix. Of the remainder, 14.8% were performed by total abdominal hysterectomy, 13.6% by Intrafascial Vaginal Hysterectomy, 12.2% by vaginal hysterectomy, and only 0.05% by Laparoscopic Assisted Vaginal Hysterectomy. Comparative data of these six surgical techniques concerning patients' characteristics, indications for operation, histological features, blood loss, operating time, hospital stay, uterine weights and postoperatively used analgesics are described. PMID- 9225649 TI - Treatment of uterine fibroids with GnRH-analogues prior to hysterectomy. AB - GnRH-analogues have a longer half-life and stronger receptor affinity than native GnRH. They block the secretion of gonadotropins from the anterior pituitary, producing a hypoestrogenic state. Preoperative treatment with GnRH-agonists for 3 months prior to hysterectomy reduces the size of uterine fibroids by about 50%. The hematologic profile is improved and subjective symptoms reduced. Other results are: a smaller peroperative blood loss, a shorter hospitalization time, and a tendency to easier operations. Side effects are predominantly hypoestrogenic. Treatment with GnRH-agonists before hysterectomy is recommended in cases of large fibroids, when technical problems may be anticipated, when preoperative anemia is present, or when it is desirable to postpone surgery. PMID- 9225650 TI - Can pre-eclampsia be predicted and prevented? AB - Pre-eclampsia is still a condition which worldwide kills many mothers and is the cause of a great number of perinatal deaths and one of the leading causes of perinatal morbidity even in developed countries. The definition of pre-eclampsia still varies in the literature, thus making the comparison of results difficult and the issue obscure. Nevertheless, recent advances in research into the pathophysiology of pre-eclampsia has greatly enhanced our understanding of his condition. Therefore, new strategies to predict and prevent this disorder are sought for and emerging. PMID- 9225651 TI - Hemodynamic measures in prediction of pre-eclampsia. PMID- 9225652 TI - Biochemical prediction of pre-eclampsia. AB - Various biochemical tests, suggested to be of value in the prediction of pre eclampsia, have been evaluated. Some of these tests are currently available for use clinical, whereas others exist only in the laboratory. None, however, has been found to fulfil all desired criteria for the prediction of the development of pre-eclampsia. PMID- 9225653 TI - Non-medical prevention of pre-eclampsia. AB - Daily supplementation with 1.5-2 g calcium is the only known effective method of non-medical prevention of pre-eclampsia. Data concerning the role of bed rest for prevention of this condition are still controversial. Fish oil and magnesium supplements have proved ineffective in clinical trials. Dietary salt restriction is not acceptable in view of its possible adverse effects. PMID- 9225654 TI - Medical prevention of pre-eclampsia. AB - There is no clear evidence that any of the antihypertensive drugs available can defer or prevent the occurrence of proteinuric pre-eclampsia or associated problems such as fetal growth retardation or perinatal death. When antihypertensive treatment is indicated, there seems to be no reason to prefer any of the tested beta-blockers, or to prefer labetalol to a pure beta-blocker, or indeed, to prefer beta-blockers to methyldopa. The increased maternal, fetal and infant mortality and morbidity associated with hypertension in pregnancy justify careful evaluation of the risks of the more severe forms of hypertension at an early stage in all pregnancies. A careful family and medical history are benchmarks in pregnancy surveillance, followed by meticulous monitoring of the pregnant mother in a well organized maternity health care system where high maternal compliance is necessary together with use of appropriate methods to predict hypertension early. Prophylactic treatment with medication causing least harm to the mother and fetus to prevent serious complication due to hypertension in pregnancy when increased risk is identified would be of value and further improve maternal and fetal outcome. PMID- 9225655 TI - Is screening for genital infections in pregnancy necessary? AB - In recent decades, cervical screening for gonorrhea has been an integral part of antenatal care. Today, in most countries in the "western world", other microorganisms commonly found in the vagina are important causes of premature labor and are associated with perinatal and puerperal infections. These include Chlamydia trachomatis, Group B streptococci, herpes simplex virus, genital mycoplasmas and bacterial vaginosis. This paper discusses current strategies to prevent complications of these infections in pregnancy and at birth. PMID- 9225656 TI - Diagnosis of infections due to Chlamydia trachomatis. PMID- 9225657 TI - Epidemiology of sexually transmitted diseases in the Baltic countries. AB - According to the UN definition the Baltic countries belong to the group of countries whose economy is in transition. This transition period has created changes in life-style, priorities and living standards. The objective of this study was to determine the incidence, distribution and control of sexually transmitted diseases (STDs) in the Baltic countries and to draw some comparisons with data from Scandinavia. We have compared the official statistical data concerning STDs from 1991-94 in all three Baltic countries and have attempted to obtain information about the way these reports are collected. We have come to the following conclusions: -the incidence rate of STD in the Baltic countries is increasing, -the average age of patients suffering from STDs is decreasing, -the specificity of the diagnostic methods used for STDs (especially Chlamydia trachomatis) needs to be improved. Facilities for diagnosing HSV and HPV should be made available. PMID- 9225658 TI - The changing face of prostitution in the Baltic sea area. PMID- 9225659 TI - HIV infection and pregnancy. PMID- 9225660 TI - Stable coronary artery disease: the family physician's approach. PMID- 9225662 TI - ADHD: the role of the family physician. PMID- 9225661 TI - A new era in HIV care. PMID- 9225663 TI - Once-daily aminoglycoside dosing. PMID- 9225664 TI - Counseling the infertile couple: when enough is enough. PMID- 9225665 TI - Mitral valve prolapse in patients with anorexia nervosa. PMID- 9225666 TI - Dying with dignity. PMID- 9225667 TI - Follow-up of the minimally abnormal Pap smear. PMID- 9225668 TI - Opportunistic infections and psychosocial stress in HIV. AB - Persons infected with the human immunodeficiency virus (HIV) are susceptible to both opportunistic infections and psychosocial crises at varying stages of their disease. Many interventions, including lifestyle changes and chemoprophylaxis, may help prevent, delay or lessen the extent of the morbidities associated with immunodeficiency. In appropriate patients, prophylaxis for infections with Pneumocystis carinii, Mycobacterium avium complex and Toxoplasma helps to reduce morbidity. PMID- 9225670 TI - Providing asthma education in primary care practice. AB - An educational dialog founded on open communication between clinician and patient is necessary for a successful partnership in asthma care. Educating patients in self-management of asthma has become a major challenge in primary care practice. The process should begin at the time of diagnosis and should be integrated into every step of medical care during office visits and any other clinician-patient interaction. Identifying patients' expectations and concerns about their disease at each office visit helps the clinician focus care. The educational effort should include a discussion of basic asthma facts, and the types and uses of medications, and a demonstration of the skills involved in the proper use of metered-dose inhalers, spacers and peak flow meters. A written asthma self management plan developed jointly by the clinician and the patient includes recommended daily medications and the specific steps the patient should take to control asthma and achieve the goals of asthma care. PMID- 9225669 TI - Management of stable coronary artery disease. AB - Options for the treatment of multivessel coronary artery disease include medical therapy and revascularization with either coronary artery bypass grafting (CABG) or percutaneous transluminal coronary angioplasty (PTCA). CABG has been shown to prolong survival in patients with left main coronary disease or when at least two of the following factors are present: extensive coronary artery disease, significant ischemia as shown on stress testing and left ventricular dysfunction. In patients with extensive coronary artery disease but normal left ventricular function, either PTCA or CABG may be used initially without adversely affecting rates of survival. However, patients undergoing angioplasty have a higher rate of repeat revascularization procedures than those treated with bypass. The complex interplay of the various risks and benefits of medical therapy, angioplasty or bypass in the treatment of coronary artery disease requires an individualized approach to therapy and careful patient education. PMID- 9225671 TI - Necrotizing fasciitis: a diagnostic challenge. AB - Group A streptococci are responsible for much of the morbidity associated with skin infections. Necrotizing fasciitis, the most extreme form of these infections, may be life-threatening. Consequently, physicians need to know how to diagnose and effectively treat this deep infection of the subcutaneous tissues. The diagnosis of necrotizing fasciitis is based on the history (i.e., predisposing factors to infection), gram staining and culture, radiography and, ultimately, surgical exploration. The infection is treated with antibiotics, hyperbaric oxygenation and surgical debridement. PMID- 9225672 TI - Evaluating the hyperactive child in your office: is it ADHD? AB - When confronted with the complaints of hyperactivity and impulsivity in a child, the family physician may find it tempting to initiate a complex process leading to the diagnosis of attention-deficit hyperactivity disorder, which is arguably the most prevalent neurobiologic disorder in childhood. While the diagnosis of attention-deficit hyperactivity disorder cannot be made solely on the basis of office visits, a practical strategy for an initial assessment of hyperactivity and impulsivity in the examination room is extremely important. Careful observation of the child's behavior and strategic questioning of the parents can be helpful in differentiating volitional misbehavior from the neurologically based lack of behavioral control in attention-deficit hyperactivity disorder. A skilled initial office assessment allows more effective use of behavioral checklists and neuropsychologic tests, increases diagnostic confidence and provides a yardstick for measuring the effectiveness of treatment. PMID- 9225673 TI - Stress and insufficiency fractures. AB - Stress fractures of the axial and appendicular skeleton typically present as localized pain that develops without a history of specific acute injury. The differential diagnosis includes primary or metastatic neoplasms, infections, musculoskeletal soft tissue injuries, nerve compression syndromes and joint diseases. Plain radiographs may demonstrate changes consistent with fracture, including a fracture line or fracture callus. In many cases, however, initial radiographs are normal or nondiagnostic. This occurs most frequently in three situations: (1) when radiographs are obtained soon after the onset of symptoms, before the appearance of a fracture line or new bone formation; (2) in patients with osteopenia, in whom detection of a fracture line and new bone formation is difficult, and (3) when the fracture involves areas of the skeleton that are difficult to study with plain films (including the spine and pelvis). When the plain films are normal, other tests such as bone scans, computed tomography or magnetic resonance imaging usually demonstrate the fracture. PMID- 9225674 TI - Pleomorphic adenoma of the parotid. AB - Pleomorphic adenoma of the parotid is the most common tumor of salivary gland origin, accounting for 60 to 70 percent of all benign salivary gland tumors. This lesion usually presents as a slow-growing painless mass inferior to the pinna of the ear. The diagnosis is based on clinical presentation, magnetic resonance imaging or computed tomography, and fine-needle aspiration biopsy. The treatment is wide excision in which the entire capsule is removed but the facial nerve is spared. Proper diagnosis and treatment are necessary to prevent the complications of tumor recurrence and malignant transformation. Carcinoma expleomorphic adenoma arises in longstanding tumors and has a five-year recurrence rate of 75 percent. PMID- 9225675 TI - Foot infections in patients with diabetes. AB - The combination of sensory neuropathy, ischemia and direct adverse effect on host defense mechanisms makes patients with diabetes vulnerable to foot infections. A high degree of clinical suspicion and vigilance is necessary for early diagnosis of soft tissue infections and their differentiation from noninfected ulcers. Diagnosis and assessment depend primarily on clinical history and physical examination, although radiographs, scans and laboratory tests may also provide useful clinical data. The ability to detect bone in the base of an ulcer with a blunt sterile probe may be particularly useful in assisting the recognition of osteomyelitis. Most non-limb-threatening infections are caused by Gram-positive cocci, but more serious infections are often polymicrobial. Effective treatment is based on a comprehensive strategy of wound care, avoidance of weight bearing, optimal metabolic control, appropriate antibiotic use and, possibly, surgical intervention. PMID- 9225676 TI - Nutrition during pregnancy. AB - Nutrition assessment and counseling are integral components of preconception and prenatal care. The average-size woman should gain between 11.25 and 15.75 kg (25 and 35 lb) during a normal pregnancy. Some factors identify the pregnant woman with a nutrition risk. Vitamin and mineral supplementation should be based on a dietary assessment. Common discomforts of pregnancy frequently can be managed with dietary modification and safe pharmacotherapeutics. The coordinated efforts of health care providers, registered dietitians, the Women, Infants, and Children (WIC) nutrition program, local health departments and Cooperative Extension Service offices can provide appropriate nutrition assessment, education and intervention. PMID- 9225677 TI - Drug interactions with the nonsedating antihistamines. AB - The nonsedating antihistamines are frequently prescribed agents. Well-documented drug-drug interactions with two of these agents, terfenadine and astemizole, may result in serious adverse effects, including death, when they are prescribed along with macrolide antibiotics and/or the antifungal agents itraconazole or ketoconazole. Fexofenadine and loratadine appear to be the least likely nonsedating antihistamines to interact with other medications and to result in a life-threatening interaction. This article reviews the known drug-drug interactions involving nonsedating antihistamines and provides a basis from which the clinician can predict potential interactions. PMID- 9225678 TI - Advisory Committee on Immunization Practices issues recommendations for the 1997 98 influenza season. PMID- 9225679 TI - Rett syndrome: a disorder affecting early brain growth. PMID- 9225680 TI - The clinical syndrome of early-life bilateral hippocampal sclerosis. AB - Four infants had bilateral hippocampal sclerosis by magnetic resonance scans, including oblique coronal fast spin echo images of the temporal lobes; [18F]fluorodeoxyglucose-positron emission tomographic scans, done in 2 infants, showed isolated bilateral anterior temporal lobe hypometabolism. All had epilepsy with episodes of status epilepticus. Despite adequate motor and sensory functions, all failed to develop language (or lost attained language), social skills, and complex purposive or adaptive activity, even after epilepsy was controlled. Bilateral hippocampal dysfunction in early life appears to be associated with a profound failure of cognitive capacities, including language learning and learning of complex social and adaptive skills in general. The deficits correspond to the cognitive deficits of severe infantile autism. Hippocampal function, or more generally medial temporal lobe function, appears necessary for language learning in the infant, as well as for complex social and adaptive learning. PMID- 9225681 TI - Cytokine profile of myelin basic protein-reactive T cells in multiple sclerosis and healthy individuals. AB - Myelin basic protein (MBP)-reactive T cells have been implicated in the autoimmune pathogenesis of multiple sclerosis (MS). In this study, we examined the cytokine profile of 531 primary MBP-reactive T-cell lines and 72 independently established clones from 32 patients with MS and 18 healthy controls (NS) by using highly sensitive enzyme-linked immunosorbent assays. An increased number of primary T-cell lines producing interferon-gamma (IFN gamma) and/or interleukin-4 (IL-4) in response to MBP were found in patients with MS compared with controls. No distinct Th1 or Th2 subtypes could be demonstrated among the MBP-reactive clones. IL-4 was more frequently observed among MS-derived clones. Clones derived from MS patients produced increased levels of IL-2, IL-4, tumor necrosis factor-alpha (TNF alpha), IFN gamma, and IL-10, but not IL-6. It is interesting that MBP-reactive T cells from MS patients expressing the disease associated HLA-DRB1*15 allele produced increased quantities of TNF alpha, a cytokine suggested to play an important role in inflammation and demyelination. When challenged with either MBP or a bacterial superantigen, the clones expressed similar levels of the proinflammatory cytokine IFN gamma. Our study suggests a functional difference in T-cell responses to MBP in patients with MS compared with healthy individuals, and provides further insights into the role of MBP reactive T cells and their cytokine profile in the inflammatory processes of MS. PMID- 9225682 TI - Association of Campylobacter jejuni serotype with antiganglioside antibody in Guillain-Barre syndrome and Fisher's syndrome. AB - Using Penner's method and Lior's scheme, we serotyped 31 isolates from patients with Guillain-Barre syndrome, 7 isolates from those with Fisher's syndrome, and isolates from patients with sporadic enteritis. PEN 19 of Campylobacter jejuni was isolated more frequently from the Guillain-Barre syndrome patients (16/31 isolates, 52%) than from the sporadic enteritis patients (5%). LIO 7 of C. jejuni also was isolated more frequently from the Gillain-Barre syndrome patients (14/31 isolates, 45%) than from the enteritis patients (3%). One reason why Guillain Barre syndrome is rare, despite the high incidence of C. jejuni enteritis, may be the low frequencies of PEN 19 and LIO 7. The frequency of positive anti-GM1 antibody titers in the Guillain-Barre syndrome patients with PEN 19 isolates was higher than that in the Guillain-Barre syndrome and Fisher's syndrome patients without PEN 19 isolates. We speculate that the serotypic determinant of PEN 19 aids in the production of anti-GM1 antibody by a GM1-like lipopolysaccharide. In contrast, 5 of the 7 isolates from the Fisher's syndrome patients belonged to PEN 2:LIO 4. The IgG anti-GQ1b antibody was associated with PEN 2 and LIO 4. These serotypic determinants may aid in the production of IgG anti-GQ1b antibody by a GQ1b-like lipopolysaccharide. PMID- 9225683 TI - Accumulation of beta-amyloid precursor protein in HIV encephalitis: relationship with neuropsychological abnormalities. AB - The pathogenesis of neuropsychological abnormalities in patients with human immunodeficiency virus type 1 (HIV-1) encephalitis is obscure because neurons are not the target of infection and severe neuronal loss occurs only late during the disease. Moreover, there is evidence indicating that HIV dementia is not a homogeneous entity and could partially reverse after treatment with zidovudine. The finding that impaired axonal flow, evidenced by beta-amyloid precursor protein immunoreactivity, could contribute to the neuropsychological deficits prompted the present study. Brains of patients with full-blown acquired immunodeficiency syndrome (AIDS) were studied and findings compared with those of normal and abnormal control subjects. The presence of HIV-1 DNA was investigated by nested polymerase chain reaction; axonal abnormalities were detected by beta amyloid precursor protein, ubiquitin immunohistochemistry, and silver staining. Accumulation of beta-amyloid precursor protein was observed in all the HIV encephalitis brains studied; the appearance of the immunostaining varied from globular structures to bundles of parallel formations. In 2 AIDS brains without pathological abnormalities, only the latter pattern was detected. The brains with trauma were strongly reactive with beta-amyloid precursor protein antibody and the different reactivity within them correlated with posttrauma survival, only globular structures being detected in the older cases. No correlation was found between the different pattern of beta-amyloid precursor protein reactivity and dementia in AIDS patients. These results show that widespread axonal injury is a constant feature in AIDS brains and suggest that it could play a role in the pathogenesis of the neuropsychological abnormalities of these patients. PMID- 9225684 TI - Extensive DNA deletion associated with severe disease alleles on spinal muscular atrophy homologues. AB - Spinal muscular atrophy (SMA) is a motor neuron disease presenting with a wide spectrum of phenotypic variations. The primary cause of most, if not all, forms of childhood-onset spinal muscular atrophy appears to be the homozygous loss of the telomeric copy of the survival motor neuron (SMNT) gene. It is interesting that approximately half of all affected patients are likewise homozygous nulls for the neuronal apoptosis inhibitory protein (NAIP) gene and a somewhat lesser fraction for the basal transcription factor, p44 subunit (BTF2p44) gene. It has been proposed that homozygous loss of SMNT is the primary cause of spinal muscular atrophy while the loss of NAIP and perhaps other genes primarily affects the severity of disease manifestation. We explored this hypothesis by evaluating the extent of gene deletions in three multigenerational families with spinal muscular atrophy exhibiting dramatic intrafamilial phenotypic variation. Using somatic cell hybrid lines to sequester individual spinal muscular atrophy homologues, we show that homologues missing several contiguous genes correlate with "severe" disease alleles and homologues missing only SMNT correlate with "mild" disease alleles. These observations support the hypothesis that phenotypic severity among the childhood-onset spinal muscular atrophies is directly correlated with the extent of disease-specific deletions. PMID- 9225685 TI - Magnetic resonance imaging in classification of congenital muscular dystrophies with brain abnormalities. AB - A survey was performed of magnetic resonance imaging (MRI) findings in 21 patients with congenital muscular dystrophy (CMD) with cerebral abnormalities to evaluate the contribution of MRI to the classification of CMD patients. In 5 patients with Walker-Warburg syndrome (WWS), MRI showed hydrocephalus due to aqueduct stenosis, generalized cerebral cortical agyric or pachygyric polymicrogyria, diffuse cerebral hemispheric white matter abnormalities, and malformations of posterior fossa structures. In 4 patients with muscle-eye-brain disease, MRI showed cortical dysplasia, but less severe than in WWS. The cerebral white matter either was normal or contained multiple focal abnormalities. Malformations of posterior fossa structures were present. Eight patients, classified as having classic merosin-deficient CMD (MD-CMD), had diffuse cerebral hemispheric white matter abnormalities, no other abnormalities. One patient with MD-CMD had only a few, focal white matter abnormalities. Three CMD patients had occipital agyria, otherwise normal gyration, multifocal or more diffuse cerebral white matter changes, and variable hypoplasia of pons and vermis. Two of the 3 patients had negative muscle merosin staining. The conclusion of the study is that MRI is an important adjunct in the classification of CMD patients. CMD with occipital agyria can be regarded as a newly recognized, separate CMD subtype. PMID- 9225686 TI - Intrinsic epileptogenesis of hypothalamic hamartomas in gelastic epilepsy. AB - Hypothalamic hamartomas and gelastic seizures are often associated with cognitive deterioration, behavioral problems, and poor response to anticonvulsant treatment or cortical resections. The origin and pathophysiology of the epileptic attacks are obscure. We investigated 3 patients with this syndrome and frequent gelastic seizures. Ictal single-photon emission computed tomography performed during typical gelastic seizures demonstrated hyperperfusion in the hamartomas, hypothalamic region, and thalamus without cortical or cerebellar hyperperfusion. Electroencephalographic recordings with depth electrodes implanted in the hamartoma demonstrated focal seizure origin from the hamartoma in 1 patient. Electrical stimulation studies reproduced the typical gelastic events. Stereotactic radiofrequency lesioning of the hamartoma resulted in seizure remission without complications 20 months after surgery. The functional imaging findings, electrophysiological data, and results of radiofrequency surgery indicate that epileptic seizures in this syndrome originate and propagate from the hypothalamic hamartoma and adjacent structures. PMID- 9225687 TI - Circulating tumor necrosis factor-alpha correlates with electrodiagnostic abnormalities in Guillain-Barre syndrome. AB - Autoimmune damage to peripheral nerves, mediated by activated T lymphocytes and macrophages, underlies the pathogenesis of inflammatory demyelination in Guillain Barre syndrome. Both T lymphocytes and macrophages secrete tumor necrosis factor alpha, a cytokine that exerts toxic effects on myelin, Schwann cells, and endothelial cells. The reportedly high serum levels of this cytokine in patients with Guillain-Barre syndrome may reflect the degree of immune activation rather than a direct pathogenic effect. We compared serum levels of tumor necrosis factor-alpha, interleukin-1 beta, and soluble interleukin-2 receptor with well established electrodiagnostic criteria for primary demyelination in 23 patients with Guillain-Barre syndrome, to assess the relationship between these cytokines and peripheral myelin damage. High serum levels of tumor necrosis factor-alpha were associated with prolonged distal motor latencies and slowed motor conduction velocities, prolonged or absent F-wave responses, and reduced amplitude of distal compound muscle action potentials. No significant correlation was observed between electrodiagnostic criteria for primary demyelination and serum levels of interleukin-1 beta or soluble interleukin-2 receptor. These findings suggest a putative role of tumor necrosis factor-alpha in the pathogenesis of peripheral nerve demyelination in Guillain-Barre syndrome. PMID- 9225688 TI - Multiple sclerosis: oligodendrocytes display cell death-related molecules in situ but do not undergo apoptosis. AB - To investigate whether apoptosis is involved in the fate of oligodendrocytes in chronic multiple sclerosis lesions, the pro-apoptotic molecules fas and tumor necrosis factor receptors and the anti-apoptotic molecule bcl-2 were examined by immunohistochemistry, and DNA fragmentation was assessed by an end labeling technique. Fas and both tumor necrosis factor receptors were preferentially expressed on oligodendrocytes in multiple sclerosis lesions, this phenotype being more evident at the lesion edge. The ligand for fasL, was constitutively present at high levels on microglia. The anti-apoptotic molecule bcl-2 was selectively expressed on oligodendrocytes in silent lesions and on astrocytes in active lesions. These molecules were also detected in control material, albeit at lower levels. In chronic active lesions, a few inflammatory cells displayed fas reactivity, whereas the majority expressed bcl-2. DNA fragmentation was found in a number of infiltrating cells and some microglia, whereas, with one possible exception, oligodendrocytes showed no evidence of apoptosis. Thus, while apoptosis is involved in the elimination of infiltrating cells, it plays little or no role in oligodendrocyte depletion in multiple sclerosis, a process that may be related to a lytic pathway. In addition, microglia constitutively displayed the ligand for fas, and appeared to be the major effector cell population in the central nervous system. PMID- 9225689 TI - Metabolic reduction in the posterior cingulate cortex in very early Alzheimer's disease. AB - This study investigated cerebral glucose metabolism in very early Alzheimer's disease, before a clinical diagnosis of probable Alzheimer's disease is possible, using [18F]fluorodeoxyglucose positron emission tomography. First, 66 patients with probable Alzheimer's disease with a spectrum of dementia severity (Mini Mental State Examination score, 0-23) were recruited and studied. Cortical metabolic activity was analyzed topographically using three-dimensional stereotactic surface projections. Regression analysis was performed for each brain pixel to predict metabolic patterns of very early disease. Predictions were tested prospectively in a group of 8 patients who complained only of memory impairment without general cognitive decline (Mini-Mental State Examination score, 25 +/- 1) at the time of scanning but whose condition later progressed to probable Alzheimer's disease. Both results were compared to cerebral metabolic activity in 22 age-similar normal control subjects. Prediction and analysis of actual patients consistently indicated marked metabolic reduction (21-22%) in the posterior cingulate cortex and cinguloparietal transitional area in patients with very early Alzheimer's disease. Mean metabolic reduction in the posterior cingulate cortex was significantly greater than that in the lateral neocortices or parahippocampal cortex. The result suggests a functional importance for the posterior cingulate cortex in impairment of learning and memory, which is a feature of very early Alzheimer's disease. PMID- 9225690 TI - Short- and long-term survival and function of unilateral intrastriatal dopaminergic grafts in Parkinson's disease. AB - Six patients with Parkinson's disease were followed for 10 to 72 months after human embryonic mesencephalic tissue from four to seven donors was grafted unilaterally into the putamen (4 patients) or putamen plus caudate (2 patients). After 8 to 12 months, positron emission tomography showed a 68% increase of 6-L [18F]-fluorodopa uptake in the grafted putamen, no change in the grafted caudate, and minor decreases in nongrafted striatal regions. There was therapeutically valuable improvement in 4 patients, but only modest changes in the other 2, both of whom developed atypical features. Patient 4 was without L-dopa from 32 months and had normal fluorodopa uptake in the grafted putamen at 72 months. Overall, the L-dopa dose was reduced by a mean of 10 and 20%, "off" time was reduced by 34 and 44%, and the "off" phase Unified Parkinson's Disease Rating Scale motor score by 18 and 26%, and the duration of the response to a single L-dopa dose increased by 45 and 58% during the first and second years after surgery, respectively. Rigidity and hypokinesia improved bilaterally, but mainly contralateral to the implant. No consistent changes in dyskinesias were observed. We conclude that transplantation of embryonic mesencephalic tissue leads to highly reproducible survival of dopaminergic neurons, inducing clinically valuable improvements in most recipients. PMID- 9225691 TI - Presenilin-1 polymorphism and hereditary cerebral hemorrhage with amyloidosis, Dutch type. AB - Hereditary cerebral hemorrhage with amyloidosis, Dutch type, caused by a mutation at codon 693 of the amyloid beta precursor protein gene, is characterized by amyloid beta deposition resulting in recurrent strokes and dementia. Recent data suggest that presenilin-1 may be biologically linked to cerebral amyloid beta deposition. The intronic presenilin-1 polymorphism published by Wragg and colleagues (1996) was analyzed in 65 carriers of the hereditary cerebral hemorrhage with amyloidosis, Dutch type, mutation. We found that the presenilin-1 genotype was not correlated with age at first stroke, number of recurrences, dementia, and age at death or with white matter hyperintensities and focal lesions on magnetic resonance images. From our data we conclude that amyloid beta deposition in this disease is most likely not influenced by presenilin-1. PMID- 9225692 TI - Sporadic focal dystonia in northwest Germany: molecular basis on chromosome 18p. AB - Idiopathic focal dystonia (IFD) is the most common form of idiopathic torsion dystonia in the Euroamerican population, with a prevalence of about 30 per 100,000. Although most patients claim a negative family history, we recently mapped this syndrome to chromosome 18p as an autosomal dominant trait in Family K from Northwest Germany. We now have investigated sporadic patients with IFD from the same geographic area both clinically and molecularly with chromosome 18p markers. The data indicate that most of these apparently sporadic patients have inherited the same mutation as Family K from a common ancestor and, in fact, owe their disease to autosomal dominant inheritance at low penetrance. The data also indicate that this dystonia mutation (DYT7) is the predominant cause of IFD, at least in this area of Northwest Germany, and that its location can be narrowed from a 30- to a 6-centimorgan region close to marker D18S1098. PMID- 9225694 TI - Selective vestibular damage in neurosarcoidosis. AB - We report a patient with neurosarcoidosis who developed bilateral benign paroxysmal positional vertigo (BPPV) of the posterior canals, deafness, and absent responses to conventional caloric and rotational vestibular testing. Additional rotation in the planes of the vertical semicircular canals revealed relative sparing of vertical canal function. This vertical-horizontal canal dissociation explains the presence of BPPV and suggests that the vestibular damage in this patient is secondary to a vasculitic neuropathy. PMID- 9225693 TI - alpha 1-Antichymotrypsin as a risk modifier for late-onset Alzheimer's disease in Japanese apolipoprotein E epsilon 4 allele carriers. AB - In the Japanese population, sporadic late-onset Alzheimer's disease (LOAD) cases had significantly higher frequencies of the A allele of alpha 1-antichymotrypsin (ACT) gene as well as the epsilon 4 allele of apolipoprotein E (APOE) gene than controls. The odds ratio for LOAD in APOE4 carriers with the ACT-A allele was more than six times that in APOE4 carriers without the ACT-A allele (21.1 vs 3.2). These results indicate that the ACT-A allele is a risk modifier for LOAD in APOE4 carriers. PMID- 9225695 TI - Polymerase chain reaction-based detection of Tropheryma whippelii in central nervous system Whipple's disease. AB - Whipple's disease of the central nervous system (CNS) may be associated with normal intestinal histology as a result of minimal or patchy involvement. The diagnosis is difficult and is frequently made post mortem. We studied 6 patients with clinically suspected CNS Whipple's disease; 2 had oculomasticatury myorhythmia (OMM) fitting criteria for a diagnosis of definite CNS Whipple's disease. One of the 2 had duodenal histology highly suggestive of Whipple's disease the other 5 patients had normal duodenal histology. DNA was extracted from paraffin-embedded duodenal tissues in all patients and frozen pontine tissue in 1. Two primer pairs (W3F-W4R, W3F-W2R) were used in separate polymerase chain reactions (PCRs) to amplify fragments of Tropberyma whippelii 16S rDNA from these tissue samples. PCR amplicons were detected only in the duodenal tissues from the 2 patients with OMM. The sequences of these amplicons were identical to the corresponding region of the previously published Tropheryma whippelii 16S rDNA sequence. PCR-based assays of intestinal or brain tissue may be of value for confirming, and possibly refuting, a clinical diagnosis of CNS Whipple's disease in a patient with any combination of dementia, supranuclear gaze palsy, hypothalamic manifestations, myoclonus, seizures, ataxia, or OMM, especially when tissue histology is unrevealing. PMID- 9225696 TI - Early-onset Alzheimer's disease with a presenilin-1 mutation at the site corresponding to the Volga German presenilin-2 mutation. AB - We describe a new mutation causing Alzheimer's disease (AD) in presenilin-1 (N135D) that is at the homologous site to the presenilin 2 mutation (N141I) in Volga German kindreds. The phenotype of PS1 N135D is an early-onset (34-38 years) disease. The mutation forms part of, and extends, the alpha-helical array of mutations in transmembrane 2 of the presenilins and leads to the suggestion that disruption of this helical face is the molecular insult that leads to disease. PMID- 9225697 TI - Levodopa-inhibiting effect of pallidal surgery. PMID- 9225698 TI - Copper and zinc levels in familial amyotrophic lateral sclerosis patients with CuZnSOD gene mutations. PMID- 9225699 TI - Acute myopathy of intensive care. PMID- 9225700 TI - Inmate access to postrelease medical care: public health implications. PMID- 9225701 TI - Rural physicians, rural networks, and free market health care in the 1990s. AB - The changes brought about by managed care in America's urban communities will have profound effects on rural physicians and hospitals. The rural health care market characterized by small, independent group practices working with community hospitals is being offered affiliations with large, often urban-based health care organizations. Health care is evolving into a free market system characterized by large networks of organizations capable of serving whole regions. Rural provider initiated networks can assure local representation when participating in the new market and improve the rural health infrastructure. Although an extensive review of the literature from 1970 to 1996 reveals little definitive research about networks, many rural hospitals have embraced networking as one strategy to unify health care systems with minimal capitalization. These networks, now licensed in Minnesota and New York, offer rural physicians the opportunity to team up with their community hospital and enhance local health care accessibility. PMID- 9225702 TI - Developing a strategy for managing behavioral health care within the context of primary care. AB - Although most patients with psychological disorders are diagnosed and treated within the primary care setting, there are few guidelines to help primary care physicians and managed care plan administrators construct programs of behavioral health care that are compatible with the primary care environment. We report the findings from a review of the literature from 1970 to 1996 on factors that predict the use of mental health and substance abuse services with specific reference to primary care. We use a heuristic framework of service use that includes the characteristics of patients, primary care physicians, practice settings, and managed care plans. Recognizing that the factors associated with the use of services center on the primary care practice, we argue that programs of behavioral health care will work best when they are decentralized to account for variations among primary care patients, physicians, and practices; when they are integrated clinically, financially, and administratively within the primary care setting; and when primary care physicians are active leaders in the design and implementation of these services, for clinical and financial reasons. PMID- 9225703 TI - The 'usual care' of major depression in primary care practice. AB - OBJECTIVE: To determine how primary care physicians treat patients with major depression in the course of routine practice and the degree to which such practice produces outcomes anticipated with interventions recommended by the Agency for Health Care Policy and Research Depression Guideline Panel. DESIGNS: Prospective cohort study. SETTINGS: Academically affiliated ambulatory family practice centers and internal medicine clinics in urban neighborhoods of Pittsburgh, Pa. PATIENTS: Ninety-two patients who were seen in primary care practices and who met criteria for a current major depression as determined by the Diagnostic Interview Schedule and a psychiatrist's assessment. INTERVENTION: Physicians were informed of the patient's psychiatric diagnosis, and were urged to treat it in whatever manner and for whatever duration they deemed appropriate (ie, with "usual care"). MAIN OUTCOME MEASURES: The treatments that were provided, the patients' clinical course, and the relationship between the type of treatment and clinical course. RESULTS: Health center records indicated that 67 patients (73%) received a depression-specific treatment in the 8 months following study entry. A majority of the total cohort were prescribed an antidepressant drug. Of the 92 patients, 18 (20%) were asymptomatic at 8 months (Hamilton Rating Scale for Depression score, < or = 7). The treatment pattern was not clearly related to the clinical course. CONCLUSIONS: The recovery rates for the patients with major depression who were treated with usual care in routine primary care practices were lower than those anticipated from treatments consistent with the Agency for Health Care Policy and Research guidelines. Further studies of the caregiving elements that influence the effectiveness of depression-specific treatments of patients in primary care settings are needed. PMID- 9225704 TI - The 'usual care' of depression is not 'good enough'. PMID- 9225705 TI - Communication of preferences for care among human immunodeficiency virus-infected patients. Barriers to informed decisions? AB - OBJECTIVE: To examine the way patients with serious, progressive illnesses communicate their care preferences to their physician. DESIGN: An observational, cross-sectional survey of 1031 clients with acquired immunodeficiency syndrome (AIDS) or symptomatic human immunodeficiency virus disease. Self-report of communication was assessed in 861 clients who stated a treatment preference focused on extending life or focused on comfort even if it shortened life. SETTING: The Robert Wood Johnson AIDS Health Services Program in 9 US cities. PARTICIPANTS: Eight hundred sixty-one of 1031 clients recruited to the AIDS Health Services Program. RESULTS: Eight hundred sixty-one subjects expressed a preferred treatment approach; however, only 35.8% had spoken to their physician about their preferred treatment. Black clients were half as likely (odds ratio, 0.49; confidence interval, 0.29-0.85) to have discussed their preferred treatment approach even after adjustment for age, function, education, income, and other covariates. Black clients were half as likely to prefer an approach to care that focused only on comfort (odds ratio, 0.51; 95% confidence interval, 0.34-0.76). Clients with AIDS who were symptomatic daily, college educated, and more functionally impaired were more likely to have discussed a preferred treatment approach with their physician. CONCLUSIONS: Most persons with symptomatic human immunodeficiency virus infection have not discussed their preferred treatment approach with a physician. This disparity is greater for blacks, who were less likely to want a palliative treatment approach. PMID- 9225706 TI - Women's triage and management preferences for cervical cytologic reports demonstrating atypical squamous cells of undetermined significance and low-grade squamous intraepithelial lesions. AB - OBJECTIVE: To determine women's triage test preferences for the evaluation and management of atypical squamous cells of undetermined significance (ASCUS) and low-grade squamous intraepithelial lesion (LSIL) Papanicolaou smear reports. DESIGN: A 35-item questionnaire. SETTINGS: Primary care clinic waiting rooms. PARTICIPANTS: A convenience sample of 968 women. INTERVENTION: Women received standardized descriptions of the meaning of ASCUS and LSIL Papanicolaou smear classifications and uniform descriptions of the 4 triage tests: Papanicolaou smear, human papillomavirus DNA test, cervicography, and colposcopy. MAIN OUTCOME MEASURES: Subjects' responses to questionnaire. RESULTS: More women (58.4%) preferred a repeat Papanicolaou smear for an ASCUS report than would choose human papillomavirus DNA testing (7.3%), cervicography (20.6%), or colposcopy (13.8%) (P < .001, chi 2). Alternatively, 51% of women wanted colposcopy to evaluate an LSIL report compared with the other 3 options (P < .001, chi 2). Test accuracy was the most important factor that influenced women's decisions for each test, compared with cost, discomfort, and other reasons (P < .001, chi 2). Positive predictors for women's selection of colposcopy to evaluate a Papanicolaou smear showing LSIL included older age (P < .01, logistic regression analysis), higher level of income (P < .001, chi 2), greater level of education (P < .001, logistic regression analysis), greater level of knowledge of colposcopy and Papanicolaou smears (P < .001, logistic regression analysis), family history of cervical cancer (P < .01, chi 2), and history of cervical dysplasia (P = .02, chi 2). CONCLUSIONS: Most women preferred a repeat Papanicolaou smear to further evaluate an initial Papanicolaou smear demonstrating ASCUS and colposcopy to evaluate a report of LSIL. Women identified test accuracy as the most important reason for triage test selection. Multiple factors, primarily involving patient and family history of cervical neoplasia, level of education, income, age, and knowledge of tests, influence women's desire for specific triage tests. Because no optimal management of women with ASCUS and LSIL Papanicolaou smear reports has been determined, consideration of women's triage test preferences should complement overall patient care. PMID- 9225707 TI - Change in coronary risk and coronary risk factor levels in couples following lifestyle intervention. The British Family Heart Study. AB - OBJECTIVES: To measure the extent to which changes in cardiovascular risk factors were correlated among married couples following a 1-year primary care, family centered, cardiovascular lifestyle intervention program and to identify couples who benefited most from this prevention program. DESIGN: Observational study. SETTING: Thirteen primary care centers in 13 towns in Britain. PARTICIPANTS: A total of 1477 men aged 40 to 59 years and their female partners who attended a family health checkup in 1991 to 1992 from randomly ordered invitations to registered families. After 1 year, 1204 (82%) partner pairs were rescreened. MAIN OUTCOME MEASURES: One-year changes in cigarette smoking, systolic blood pressure, serum cholesterol level, blood glucose level, and a total coronary risk score. RESULTS: Comparing men and women partners, baseline values and 1-year changes in overall coronary risk score (Pearson r = 0.27 and r = 0.20, respectively), cigarette smoking, body mass index, systolic blood pressure, cholesterol levels, and glucose levels were all positively correlated (all P < .001 except smoking cessation, P = .03). Changes in cholesterol levels and systolic blood pressure were also associated with partner's baseline measurement (P < or = .01 in both men and women). CONCLUSIONS: Men and women who benefit most from risk factor reductions have partners who also tend to benefit most. Conversely, men and women who enjoy little or no benefit have partners who tend to have similarly small benefits. It is likely that lifestyle intervention targeted at men and women as couples rather than as individuals may result in a greater reduction in cardiovascular risk factors, possibly through mutual reinforcement of lifestyle changes. PMID- 9225708 TI - Family-centered preventive counseling for coronary heart disease risk factors. Is it time for a randomized clinical trial? PMID- 9225709 TI - Effects of a systematic approach to tobacco cessation in a community-based practice. AB - Studies suggest that the absence of a systematic approach is a barrier to the provision of tobacco cessation counseling services in clinical practice. A systematic intervention was shown to be feasible and effective at a faculty practice site. Our pilot study examined the feasibility of implementation at a community-based practice and assessed the effect of the tobacco cessation counseling system on our patients' smoking behavior. Systematic assessment (smoking status, "readiness to quit"), brief counseling at each visit, and follow up (for those ready to quit) were provided by a physician and nurse team. Our results suggest that the office-based tobacco cessation counseling system can work in a community-based practice and is an effective strategy for helping smokers quit and in preparing to quit. PMID- 9225710 TI - Treatment with buspirone in a patient with autism. AB - This study evaluates the safety and efficacy of buspirone hydrochloride for the treatment of a patient with autism and hyperactivity disorder and determines the effect of buspirone on the number of performance tasks completed by the patient at school. A 3-week, double-blind, placebo-controlled crossover study was performed in a private physician, office-based practice. A child with autism, which was diagnosed by Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised, criteria, was studied. The child received placebo for 3 weeks and buspirone for 3 weeks; there was a 1-week interval between the 2 treatments. The outcome was measured by using Conners abbreviated parent and teacher questionnaires and by determining the number of daily performance tasks completed by the child at school. Statistical analysis was performed by linear models and standard F tests. Buspirone was found to be safe and efficacious, without side effects, for decreasing hyperactivity and increasing completed performance tasks. The beneficial effects of buspirone in helping this patient with autism in his natural daily settings suggest that buspirone may be an alternative to neuroleptic agents in the medical therapy of autism; further study in other patients is needed. PMID- 9225711 TI - The pharmacologic treatment of anxiety and depression in African Americans. Considerations for the general practitioner. AB - A growing pool of recent research points to the importance of ethnicity in psychopharmacologic management of depression and anxiety disorders, with sometimes profound implications for efficacy and safety. Such research has provided provocative findings that illustrate important interethnic pharmacogenetic, pharmacokinetic, and pharmacodynamic differences, especially for African Americans. We did a systematic literature review of psychopharmacologic treatment considerations among African Americans with anxiety and mood disturbance seen by primary care physicians, who provide most psychopharmacologic treatment. The findings commonly point to a greater percentage of "poor metabolizers" among African Americans compared with Euro-Americans. General treatment considerations include greater attention to adverse effects and better clinical response and poorer compliance for a given dose, potential need for lower starting doses and slower increases, use of plasma drug levels if available, determination of past responses to a similar drug, and integration of pharmacogenetic information into an overall socioculturally and ethnically sensitive approach to assessment and treatment. PMID- 9225712 TI - The injured shoulder. Primary care assessment. AB - Shoulder problems are the second most common orthopedic complaint in primary care medicine. The range of motion, ligamentous and muscular support, and central location of the shoulder are key factors for the successful performance of persons at work or on the playing field. These special attributes also contribute to injury and to difficulty in assessing the painful shoulder. An understanding of the pertinent anatomic structures, the differential diagnosis of shoulder pain (intrinsic and referred pains), and a systematic approach to the evaluation including a complete history and physical examination are necessary in this assessment. Adequate examination consists of inspection, muscle strength and range-of-motion testing, palpation, and neurologic testing of the shoulder, neck, and elbow followed by special tests to detect impingement, instability, or tendinosis. This basic assessment is augmented by the proper use of radiographs, arthrography, computed tomography, ultrasonography, and magnetic resonance imaging. An adequate database and proper assessment of the injured shoulder allow the primary care physician to make a pathoanatomic diagnosis and formulate an appropriate treatment plan and make appropriate use of orthopedic consultants. PMID- 9225713 TI - A guide to the isolated dilated pupil. AB - The poorly reactive and dilated pupil observed in a comatose patient is often thought to represent an acute third nerve palsy owing to brain herniation or aneurysm. In the well patient, however, the isolated dilated pupil is unlikely to be owing to a third nerve palsy. It is more commonly owing to other benign causes such as local iris sphincter abnormalities, pharmacologic dilation, tonic pupil syndrome, or sympathetic irritation. This article presents a diagnostic flowchart to help the primary care physician analyze this problem and prevent costly and unnecessary imaging of these patients. PMID- 9225714 TI - Are we all quacks? PMID- 9225715 TI - Another view of GH neuroregulation: lessons from the sheep. PMID- 9225716 TI - Transgenic animal models in reproductive endocrine research. PMID- 9225717 TI - Are only certain hypothyroid subjects predisposed to raised intraocular pressure? PMID- 9225718 TI - Graves' ophthalmopathy: what is the initial abnormality? PMID- 9225719 TI - Less can be more--at least in mice: osteocalcin deficiency associated with increased bone formation. PMID- 9225720 TI - Hyperactive channels and inherited hypertension: Liddle's syndrome--an epithelial sodium channelopathy. PMID- 9225721 TI - More clues from fat mice: leptin acts as an opponent of the hypothalamic neuropeptide Y system. PMID- 9225722 TI - Reversible increase of intraocular pressure in subclinical hypothyroid patients. AB - OBJECTIVE: The aim of the study was to analyse the relationship between the ocular parameters, namely intraocular pressure (IOP), and the early forms of subclinical hypothyroidism. DESIGN: Fifty-three subjects (9 male and 44 female) aged from 18 to 45 years (mean 32 +/- 7 years) were selected for this study. Twenty-nine met the criteria of subclinical hypothyroidism and 24 euthyroid subjects, age- and sex-matched, were used as controls. METHODS: All individuals underwent a complete ocular examination, including visual field examination and serial measurement of IOP by means of a Goldmann tonometer. A tonographic examination was also performed. RESULTS: The hypothyroid patients showed a substantially higher pressure in both eyes compared with control subjects (right eye = 17.52 +/- 4.74 vs 13.42 +/- 1.95 mmHg, P < 0.0001; left eye = 17.55 +/- 3.99 vs 13.71 +/- 1.55 mmHg, P < 0.0001). Indeed, the tonometric pressure exceeded 18 mmHg in 11 out of the 29 (38%) patients in the right eye and in 8 out of 29 (27%) patients in the left eye. The outflow index was normal in all subjects except in two hypothyroid patients. After two months of L-thyroxine (L T4) replacement therapy, only one patient continued to show tonometric values above 18 mmHg and the hypothyroid patients showed a significant reduction in mean IOP in both eyes compared with pre-treatment values (right eye = 14.96 +/- 1.32 mmHg, P < 0.0097; left eye = 15.03 +/- 1.38 mmHg, P < 0.0018). Treatment did not lead to any change in the outflow indices; however, the C value (outflow coefficient at the sclerocorneal corner) returned to normal in the two patients with increased pre-treatment tonographic values. CONCLUSIONS: These findings indicate that the intraocular pressure is increased even in subclinical hypothyroid patients and that, at this early stage, the impairment is fully reversible with L-T4 therapy. PMID- 9225723 TI - Analysis of human thyrotropin receptor gene expression and immunoreactivity in human orbital tissue. AB - The human thyrotropin receptor (hTSHR) represents an autoantigen that plays a central role in the hyperthyroidism of Graves' disease (GD). hTSHR transcripts have recently been detected by reverse transcription (RT)-PCR in various extrathyroidal tissues, suggesting that the hTSHR may be more widely distributed than previously thought, and that it may serve as a common antigen in the thyroidal and extrathyroidal manifestations of GD. Using techniques other RT-PCR, we examined whether RNA encoding hTSHR and hTSHR-specific immunoreactivity can be detected and localized in cultured orbital fibroblasts (OF), orbital connective tissue, extraocular muscle and various extraorbital tissues derived from both patients with Graves' ophthalmopathy (GO) and normal individuals. Using in situ hybridization with a digoxigenin-labeled antisense oligonucleotide probe specific for the extracellular domain of hTSHR, specific perinuclear and cytoplasmic hTSHR gene expression was detected in OF of patients with GO and, to a lesser degree, normal individuals. Using a highly sensitive immunostaining technique and a panel of monoclonal and polyclonal antibodies directed against different epitopes of recombinant hTSHR, distinct cytoplasmic hTSHR-like immunoreactivity was detected in methanol-fixed OF and orbital connective tissue, which was absent in abdominal fibroblasts, or when using isotype-matched non-immune immunoglobulins. Mouse monoclonal and pig polyclonal hTSHR antibodies detected cytoplasmic hTSHR-like immunoreactivity in perimysial fibroblasts within extraocular muscle, but not in extraocular muscle fibers. Immunocytochemical staining with rabbit polyclonal hTSHR antibody revealed, in addition, distinct cell surface-associated immunoreactivity in paraformaldehyde-fixed OF, but not in abdominal fibroblasts. Taken together, our results suggest that RNA encoding hTSHR is present and actively processed to immunoreactive hTSHR-like protein in OF residing within orbital connective tissue and extraocular muscle. These data support the concept that OF expressing intact or variant hTSHR may act as extrathyroidal targets for sensitized T-cells and immunoglobulins in GD. PMID- 9225724 TI - Effect of natural menopause on serum levels of IGF-I and IGF-binding proteins: relationship with bone mineral density and lipid metabolism in perimenopausal women. AB - The present study was performed to examine the effect of natural menopause on serum levels of IGF-I, IGF-binding protein (IGFBP)-2 and -3 as well as on bone mass and lipid metabolism in perimenopausal women. One hundred and twenty-one healthy Japanese women, who were 45-55 years old, were studied (71 premenopausal and 50 postmenopausal women 1-9 years after menopause). Bone mineral density (BMD) was measured at the middle third of the radius by using dual energy X-ray absorptiometry. Serum levels of IGF-I, but not those of IGFBP-2 or -3, were significantly reduced in the postmenopausal group compared with the premenopausal group. One year after menopause, serum IGF-I levels were significantly lower, and the biochemical markers of bone turnover, such as serum total alkaline phosphatase level and urinary calcium to creatinine ratio, were significantly higher than the premenopausal levels. Serum levels of IGF-I, but not those of IGFBP-2 or -3, were positively correlated with BMD. Serum levels of IGFBP-2, but not those of IGF-I or IGFBP-3, were negatively correlated with body mass index and body weight. Finally, serum levels of IGFBP-3, but not those of IGF-I, were positively correlated with serum levels of total cholesterol and triglyceride. The present findings suggest that a rapid decrease in serum IGF-I levels after menopause might be partly involved in bone loss following gonadal failure and that IGFBP-2 and -3 might be related to the regulation of body mass and lipid metabolism during perimenopause respectively, although the mechanisms remain unknown. PMID- 9225725 TI - Recombinant human gonadotropins stimulate steroid and inhibin production in human granulosa cells. AB - OBJECTIVE: Recently pure gonadotropins have become available through recombinant technology. In parallel with ongoing clinical trials it is important to examine the effects of these new gonadotropin preparations in experimental studies in human granulosa cells. In the present study the effects of recombinant FSH (rFSH) and LH (rLH) on steroid and inhibin production were examined in human granulosa cells in culture. PATIENTS AND METHODS: Granulosa cells were obtained during the follicular phase of the menstrual cycle in seven women undergoing gynecological laparotomy and from follicles in stimulated cycles in women undergoing oocyte retrieval in connection with in vitro fertilization/embryo transfer. The granulosa cells were cultured in modified Medium 199 containing 1% fetal bovine serum for 4-8 days with and without hormones. Media were changed on alternate days and stored at -20 degrees C until analyzed for estradiol, progesterone and inhibin. RESULTS: Granulosa cells from natural cycles were highly responsive to rFSH which caused a dose-related (rFSH 0.1 to 100 ng/ml) increase in estradiol and progesterone accumulation. The maximal stimulatory effect was reached with a concentration of rFSH between 1 and 10 ng/ml. Granulosa cells from stimulated cycles responded highly to rLH in terms of increased progesterone production during the whole culture period. A maximal stimulatory effect was observed with rLH at a concentration of 0.1 ng/ml. Both types of granulosa cells responded to recombinant gonadotropins in terms of increased inhibin production. CONCLUSIONS: The present study demonstrates that granulosa cells from human ovarian follicles are highly responsive to recombinant gonadotropins as demonstrated by increased steroid and inhibin production. PMID- 9225727 TI - Gonadal dysfunction in males with prolactinoma: from functional modification to irreversible damage? PMID- 9225726 TI - Ovarian 17 alpha-hydroxyprogesterone responses to GnRH analog testing in oligomenorrheic insulin-dependent diabetic adolescents. AB - OBJECTIVE: To investigate the pituitary-ovarian function in adolescent girls with insulin-dependent diabetes mellitus (IDDM). DESIGN: Clinical case-control study. METHODS: The GnRH analog leuprolide acetate was administered subcutaneously to 16 adolescents with IDDM (seven eumenorrheic and nine oligomenorrheic) and 13 controls between 0800 and 0900 h. Blood samples were collected at baseline and 0.5, 3, 6 and 24 h after leuprolide to measure levels of gonadotropins, 17 alpha hydroxyprogesterone (17-OHP), androgens and estradiol. RESULTS: Mean baseline serum LH levels were significantly higher in eumenorrheic compared with oligomenorrheic IDDM patients, while peak LH responses to GnRH analog testing were similar in all subjects. Oligomenorrheic IDDM girls showed, as a group, a distinct 17-OHP response to GnRH analog stimulation, which in five out of nine girls was in the range of functional ovarian hyperandrogenism (> or = 8.6 nmol/l). Androgen and estradiol levels were not significantly altered in any group. No correlation was found between steroid levels and HbA1c levels, although the latter were significantly higher in oligomenorrheic than in eumenorrheic patients. CONCLUSION: About 50% of the oligomenorrheic IDDM adolescents had an increased ovarian 17-OHP response to GnRH analog stimulation in the range of functional ovarian hyperandrogenism. Factors other than metabolic control, such as stress, may play an etiologic role in IDDM ovarian dysfunction. PMID- 9225728 TI - Melatonin receptors are present in adult rat Leydig cells and are coupled through a pertussis toxin-sensitive G-protein. AB - Previous studies have suggested that melatonin (MLT) acts directly on rat Leydig cells by modulating androgen production. In the present study, the site of action of MLT was investigated. The binding of 2-[125I]iodomelatonin (125I-MLT; 7-240 pmol/l) to Leydig cell membrane fragments was tested in the presence or absence of guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S; 50 mumol/l). Saturation studies and Scatchard analysis revealed the existence of a high-affinity binding site with a Bmax of 46.70 +/- 3.50 fmol/mg protein and a Kd of 88.70 +/- 6.20 pmol/l; treatment with GTP-gamma-S reduced the concentration of 125I-MLT binding sites (Bmax 34.03 +/- 4.50), while increasing the Kd to 106.5 +/- 2.61 pmol/l. Pretreatment of the cells with pertussis toxin (PTX; 10 ng/ml for 16 h) resulted in a decreased binding of 125I-MLT and a lack of effect of GTP-gamma-S. Moreover, the effect of MLT on testosterone secretion induced by LH (30 mIU/ml), forskolin (1 mumol/l) and LHRH (100 nmol/l) was studied after 3-h incubation of cells which had been precultured with or without PTX. The inhibition of testosterone secretion due to MLT administration was eliminated by PTX pretreatment during forskolin and LH, but not during LHRH administration. However, 17 hydroxyprogesterone levels were higher in all groups incubated in the presence of MLT, irrespective of PTX pretreatment. Our data suggest that: (a) MLT receptors are present on the membranes of adult rat Leydig cells; (b) they couple through PTX-sensitive G-protein-coupled binding sites; (c) the mechanism by which MLT blocks 17-20 desmolase enzymatic activity (thus leading to increased 17 hydroxyprogesterone levels), and testosterone secretion during LHRH stimulation is likely to depend on one or more different mechanism(s) of action. PMID- 9225729 TI - Expression of parathyroid hormone-related peptide (PTHrP) and PTH/PTHrP receptor in newborn human calvaria osteoblastic cells. AB - We examined the expression of parathyroid hormone-related peptide (PTHrP) and its receptor in normal newborn human calvaria osteoblastic (NHCO) cells. Northern blot analysis showed that NHCO cells express a single 1.6 kb transcript of PTHrP, which was increased within 1 h (2x) and peaked at 6 h (7x) after serum treatment. In the culture media, the release of PTHrP peptide was maximally increased (4x) 24 h after the addition of serum, as determined by immunoradiometric assay. NHCO cells exhibited a cytoplasmic immunostaining for PTHrP in the presence of serum, and most PTHrP-positive cells were alkaline phosphatase-negative, suggesting that PTHrP was expressed in undifferentiated cells. Furthermore, RT-PCR analysis showed that both PTHrP and PTH/PTHrP receptor were expressed in NHCO cells in basal conditions or after stimulation with serum. The maximal PTHrP expression induced by serum suppressed PTH/PTHrP receptor expression, suggesting that PTHrP down-regulated its receptor in NHCO cells. Treatment with 10 nM human PTH(1-34) which binds to PTH/PTHrP receptors, increased intracellular cAMP levels and alkaline phosphatase activity, and decreased cell growth, indicating that ligand binding to PTH/PTHrP receptors regulates NHCO cell proliferation and differentiation. The expression and synthesis of PTHrP and the presence of functional PTH/PTHrP receptors suggest a possible paracrine mechanism of action of PTHrP in normal human calvaria osteoblastic cells. PMID- 9225730 TI - Expression of nitric oxide synthase III in human thyroid follicular cells: evidence for increased expression in hyperthyroidism. AB - Nitric oxide mediates a wide array of cellular functions in many tissues. It is generated by three known isoforms of nitric oxide synthases (NOS). Recently, the endothelial isoform, NOSIII, was shown to be abundantly expressed in the rat thyroid gland and its expression increased in goitrous glands. In this study, we analyzed whether NOSIII is expressed in human thyroid tissue and whether levels of expression vary in different states of thyroid gland function. Semiquantitative RT-PCR was used to assess variations in NOSIII gene expression in seven patients with Graves' disease, one with a TSH-receptor germline mutation and six hypothyroid patients (Hashimoto's thyroiditis). Protein expression and subcellular localization were determined by immunohistochemistry (two normal thyroids, five multinodular goiters, ten hyperthyroid patients and two hypothyroid patients). NOSIII mRNA was detected in all samples: the levels were significantly higher in tissues from hyperthyroid patients compared with euthyroid and hypothyroid patients. NOSIII immunoreactivity was detected in vascular endothelial cells, but was also found in thyroid follicular cells. In patients with Graves' disease, the immunostaining was diffusely enhanced in all follicular cells. A more intense signal was observed in toxic adenomas and in samples obtained from a patient with severe hyperthyroidism due to an activating mutation in the TSH receptor. In multinodular goiters, large follicles displayed a weak signal whereas small proliferative follicles showed intense immunoreactivity near the apical plasma membrane. In hypothyroid patients, NOSIII immunoreactivity was barely detectable. In summary, NOSIII is expressed both in endothelial cells and thyroid follicular cells. The endothelial localization of NOSIII is consistent with a role for nitric oxide in the vascular control of the thyroid. NOSIII expression in thyroid follicular cells and the variations in its immunoreactivity suggest a possible role for nitric oxide in thyrocyte function and/or growth. PMID- 9225731 TI - Role of the renin-angiotensin system in the development of thyroxine-induced hypertension. AB - OBJECTIVE: We evaluated the influence of chronic blockade of the renin angiotensin system on hypertension induced by long-term thyroxin (T4) administration. To this end, we determined the effects of chronic treatment with captopril on blood pressure, cardiac hypertrophy and other renal and metabolic variables of hypertensive hyperthyroid rats. METHODS: T4 was administered s.c. at 0.38 mumol/kg per day and captopril was given in the drinking water (1.38 mmol/l). Both treatments were maintained for 6 weeks. Control rats received tap water. After the treatment period, the rats were placed in metabolic cages. Later, blood pressure was measured in conscious rats by intra-arterial determination. RESULTS: T4-treated rats showed an increased mean arterial pressure (MAP) whereas, in rats treated with T4 plus captopril, MAP was similar to that of the control group. Captopril did not affect the increased heart rate or ventricular weight/body weight ratio of hyperthyroid rats, but it improved the reduced creatinine clearance of these animals. CONCLUSIONS: The elevation in blood pressure produced by long-term T4 administration was prevented by chronic blockade of the renin-angiotensin system. Captopril improved the renal function of hyperthyroid rats, but did not affect the relative cardiac hypertrophy of these animals. PMID- 9225732 TI - Efficacy of nonfetal human RPE for photoreceptor rescue: a study in dystrophic RCS rats. AB - This study determines the efficacy of nonfetal human retinal pigment epithelium (RPE) for photoreceptor rescue utilizing the dystrophic RCS rat as an animal model. Eyes from 10- and 49-year-old donors were obtained through the Rochester Eye and Human Parts Bank. The RPE was isolated by enzymatic treatment of the choroid-RPE with 2% dispase for 30 min at 37 degrees C. Mechanically dissociated RPE cells were injected at the superior hemisphere into the subretinal space of dystrophic RCS rats during the fourth postnatal week. Rats receiving vehicle injection served as sham controls. The animals were immunosuppressed with daily cyclosporine injections (10 mg/kg) and sacrificed 30 days posttransplantation for histologic evaluation of the RPE graft and its effect on photoreceptor survival. Transplantation of adult human RPE promoted the survival of photoreceptors in the dystrophic RCS rat. Morphometric analysis of the grafted superior hemisphere demonstrated a threefold increase in photoreceptor cell density (149.2 +/- 50 SD) compared to sham controls (39.7 +/- 31 SD) and the untouched inferior hemisphere (52.8 +/- 28 SD). RPE from the 49-year-old donor was as effective as RPE from the 10-year-old donor in promoting photoreceptor survival. The results of this study in RCS rats suggests that RPE from adult human donors of varied ages is suitable for transplantation and retains the capability to promote survival of photoreceptor cells. This finding opens the possibility of using nonfetal RPE cells in human retinal transplantation. PMID- 9225733 TI - Axotomy increases the expression of glucose-regulated protein 78 kDa in rat facial nucleus. AB - Nerve injuries lead to metabolic and morphological changes in the cell bodies of the neurons of origin. Increases in glucose turnover in axotomized facial and hypoglossal motor nuclei have been described. Glucose-regulated protein 78 kDa (GRP78) is implicated in cellular protein folding and subunit assembly and responds to glucose deficiency. We performed Western blot and immunohistochemistry to determine the effect of axotomy on the expression and regulation of GRP78 in the facial nucleus (FN). Facial nerve axotomy caused a larger and longer increase of GRP78 in the ipsilateral FN than in the contralateral FN. In right ipsilateral FN, axotomy resulted in elevation of GRP78 protein levels, first detected at 12 h and which reached significant, maximal induction at 24 h (75 +/- 27% increase). GRP78 protein levels decreased at later time points, but remained elevated over sham-operated controls. In contrast, no significant increase in GRP78 concentrations was found in contralateral left FN. Immunocytochemically, positive GRP78 staining was found mainly in the cytoplasm of motoneurons; there was no nuclear staining. Prominent GRP78-immunostaining appeared in axotomized motoneurons at 24 h postaxotomy as compared with the contralateral, unoperated controls. This augmentation was also observed at 4 and 7 days postaxotomy. The possibility that glucose metabolism and GRP78 levels are two parallel events in the injured facial nucleus is discussed. PMID- 9225735 TI - Calretinin-immunoreactive dopaminergic neurons from embryonic rat mesencephalon are resistant to levodopa-induced neurotoxicity. AB - Levodopa, which is used in the treatment of Parkinson's disease, has known cytotoxic effects on dopaminergic neurons grown in culture. Calretinin (CR) is a cytosolic calcium-binding protein found in specific subpopulations of neurons as well as in some nonneuronal tissue. CR is expressed in 10% of rat embryo dopaminergic neurons grown in vitro. Since it has been postulated that CR provides neuroprotection due to its calcium-binding properties, we investigated whether CR-containing dopaminergic neurons were spared from levodopa toxicity. Incubation of mesencephalic cells with 10(-5) to 10(-7) M levodopa on Days 1-6 in vitro produced no significant effects on the number of dopaminergic neurons containing CR, but resulted in the loss of approximately 65% of the dopaminergic cells which did not contain CR. The remaining CR-negative dopaminergic neurons exhibited dose-dependent reductions in neurite length. The neuronal processes in CR-containing dopaminergic cells retained a smooth bipolar appearance. CR immunoreactive cells which did not contain dopamine showed slight neurite length decreases at the highest drug concentrations but no changes in neuron number. These results indicate that CR may protect dopaminergic neurons from levodopa induced toxicity. PMID- 9225734 TI - FAC1 expression and localization in motor neurons of developing, adult, and amyotrophic lateral sclerosis spinal cord. AB - In this study we report the localization and expression of FAC1 protein in developing, normal adult and amyotrophic lateral sclerosis (ALS) lumbar spinal cord. High levels of FAC1 protein were detected in cells throughout all areas (gray and white matter) of the developing lumbar spinal cord. FAC1 protein was localized predominately in nuclei and the cell body of motor neurons during early stages of spinal cord development. In contrast, low levels of FAC1 protein were observed in the adult lumbar spinal cord, localized only in the cell body of large alpha motor neurons found in lamina IX. Interestingly, FAC1 protein expression was elevated in surviving motor neurons of ALS spinal cord compared to the controls and was located both in the nucleus and throughout the cytoplasm of motor neurons. FAC1 protein was also observed in white matter cells and fibers in ALS spinal cord. In support of the immunocytochemical results, in situ hybridization studies demonstrated that FAC1 mRNA is also elevated in ALS spinal cord motor neurons. These data describe the developmental regulation of FAC1 protein in the spinal cord by immunocytochemical techniques and provide evidence that this protein is reexpressed in ALS motor neurons. PMID- 9225737 TI - Cyclosporin A retards the wallerian degeneration of peripheral mammalian axons. AB - The distal (anucleate) segments of mammalian peripheral axons typically undergo complete Wallerian degeneration within 1-3 days after severance from their cell bodies, unlike invertebrates and lower vertebrates, where anucleate axons do not degenerate for weeks to months. This rapid Wallerian degeneration in mammals could be due to a more efficient immune system and/or to differences in calcium dependent pathways relative to invertebrates and lower vertebrates. To suppress the immune system and to inhibit calcium-dependent pathways in axons, we gave daily subcutaneous injections of cyclosporin A (CsA: 10 mg/kg) to Sprague-Dawley rats for 7 days before, and 5 days after, severing their right ventral tail nerves. To confirm that CsA suppressed the immune system, white blood cell density was measured in CsA-treated and in non-treated rats. Our data showed that the number of surviving anucleate myelinated axons at 5 postoperative days in CsA treated rats was significantly higher than the number in non-treated rats. Anucleate unmyelinated axons in the ventral tail nerve also exhibited better survival in CsA-treated rats than in nontreated rats. These results are consistent with the hypothesis that the immune response and/or calcium-dependent pathways play important roles in the rapid Wallerian degeneration of anucleate mammalian axons. PMID- 9225736 TI - Expression of blood-brain barrier characteristics following neuronal loss and astroglial damage after administration of anti-Thy-1 immunotoxin. AB - In most regions of the CNS, vascular endothelial cells play an important role in maintaining the composition of the neuronal microenvironment by virtue of their blood-brain barrier (BBB) characteristics. The maintenance of the endothelial BBB phenotype in vitro has been attributed primarily to astrocytes but little attention has been paid the potential role of neurons. In this study we have attempted to injure or destroy neurons and fibers of passage in a circumscribed area while leaving vascular and glial elements intact in order to determine if neurons are involved in BBB maintenance in situ. The immunotoxin OX7-SAP, a conjugate of the Thy-1 antibody OX7 and the ribosome-inactivating protein saporin, was injected into the adult rat striatum to effect neuronal death at the injection site. Although neurons and fibers of passage were destroyed within the lesion, glial cells unexpectedly were also severely injured as determined by immunohistochemical expression of several neuronal and astroglial marker proteins and ultrastructural analysis. The microvasculature remained intact, allowing a qualitative immunohistochemical analysis of several BBB markers at time points ranging from 3 to 28 days postinjection. Despite the loss of both neurons and astroglia within the lesions, the microvasculature continued to express the brain type endothelial glucose transporter GLUT-1 at all time points examined. In contrast, the barrier to endogenous protein (rat serum albumin) and the expression of endothelial barrier antigen (EBA) decreased initially but recovered even in areas that contained minimal numbers of astroglia and neuronal elements. We conclude that intact neuronal or glial cells do not appear to be necessary for the maintenance in situ of the BBB properties examined herein. PMID- 9225738 TI - In vivo hypoxia-induced neuronal damage with an enhancement of neuronal nitric oxide synthase immunoreactivity in hippocampus. AB - Although it is well known that brain ischemia is dominantly caused by hypoxia and hypoglycemia, it is still unclear how hypoxia participates in ischemia. We studied the changes in neuronal nitric oxide synthase (nNOS) and the effect of the NOS inhibitor NG-nitro-L-arginine (NNA) on hypoxia. In vivo hypoxia (5% O2/95% N2 for 30 min) induced mild degenerative neuronal changes (shrunken and eosinophilic somata with picnotic nuclei) in neurons of the CA3, the hilus of the dentate gyrus (DG) and the DG, but not in the CA1. At 3 and 7 days after hypoxia, levels of nNOS protein were significantly enhanced to 153 and 209%, but iNOS protein could not be detected. The numbers of nNOS-immunopositive neurons were significantly enhanced to 145 and 191% in the CA3, 145 and 178% in the hilus of the DG, and 243 and 387% in the DG after 3 and 7 days, respectively. In contrast, no statistical difference was determined in the CA1. We further examined the effect of NNA administered at 5 min and 3, 6, and 24 h after hypoxia. Administration of NNA (0.1 and 1 mg/kg, i.p.) significantly decreased the number of damaged neurons in the hilus of the DG and the DG. However, higher doses of NNA (10 mg/kg, i.p.) did not prevent damage. These results suggest that hypoxia alone induces enhancement of nNOS protein and nNOS immunoreactivity in neurons of the hippocampus and that NNA has biphasic effects against hypoxia-induced neuronal damage in the hilus of the DG and the DG. PMID- 9225739 TI - Calretinin-containing neurons in trimethyltin-induced neurodegeneration in the rat hippocampus: an immunocytochemical study. AB - The present study uses immunocytochemistry to investigate the behavior of the calretinin (CR)-containing neuronal subpopulation (interneurons) of the rat hippocampus in neurodegenerative processes induced by the neurotoxicant trimethyltin. Cell counts of CR-immunolabeled interneurons indicated that these cells are spared by the neurotoxicant-induced degeneration, characterized by a generalized neuronal loss, as shown by quantitative analysis after cresyl violet staining. PMID- 9225740 TI - Perinatal asphyxia induces long-term changes in dopamine D1, D2, and D3 receptor binding in the rat brain. AB - We have investigated the long-term effects of 15-16 min or 19-20 min of perinatal asphyxia on D1, D2, and D3 receptors (analyzed by quantitative autoradiography) in the mesotelencephalic dopamine systems of the 4-week-old rat. Perinatal asphyxia reduced D1 antagonist binding ([3H]SCH 23390 in the presence of ketanserine) in the accumbens nucleus, the olfactory tubercle, and the substantia nigra and increased D1 agonist binding ([3H]dopamine in the presence of spiperone) in the accumbens nucleus and the olfactory tubercle. No changes in D2 antagonist binding ([123]iodosulpride) were found, whereas D2 agonist binding ([3H]N-propylnorapomorphine, [3H]NPA) was reduced in the posterior part of the caudate-putamen, and following 19-20 min of asphyxia it was also reduced in the accumbens nucleus. D3 agonist binding (R/S-(+/-)-2-(N,N-di[2,3(n)-3H] propylamino)-7-hydroxy-1,2,3,4-tetrahydronaphthalene, [3H]7-OH-DPAT) was increased in the anterior part of the caudate-putamen following 15-16 min but not 19-20 min of asphyxia. The results indicate that perinatal asphyxia reduced the number of D1 receptors and increased D1 agonist affinity in the accumbens nucleus and the olfactory tubercle and reduced the number of D1 receptors in the substantia nigra. The number of D2 receptors was unchanged by asphyxia, whereas the D2 agonist affinity was reduced in the caudate-putamen and in the accumbens nucleus. D3 agonist binding was increased in the caudate-putamen selectively following 15-16 min of asphyxia. In conclusion, asphyxia during birth induces long-term changes in the binding characteristics of dopamine receptors in the mesotelencephalic dopamine systems, which may contribute to previously reported behavioral changes. PMID- 9225741 TI - Effects of extracellular matrix components on axonal outgrowth from peripheral nerves of adult animals in vitro. AB - Relatively little is known of the growth requirements for regenerating axons of the peripheral nervous system of adult animals. In the present study, we show that extracellular matrix material secreted by the Engelbreth-Holm-Swarm tumor cell line (matrigel) supports axonal growth from explanted peripheral nerve dorsal root ganglia (DRG) preparations of adult mice and amphibia in serum-free media, without addition of growth factors. Axonal growth in matrigel was much more profuse than that in the more commonly used gels of type 1 collagen and, after some days in culture, was accompanied by migration of Schwann cells along axons. The most abundant protein in matrigel is laminin, which has been shown in many studies to support axonal growth but, surprisingly, antisera to laminin did not inhibit axonal growth in matrigel. To determine the ability of the major components of matrigel, laminin, type IV collagen, and heparan sulfate proteoglycan (HSPG), to support axonal growth, these proteins were added to preparations of mouse peripheral nerve-DRGs in type I collagen gels. Regenerating axons were significantly longer in the presence of laminin and type IV collagen than in control cultures, while HSPG had a slight inhibitory effect. In this assay system, however, diluted matrigel solution was even more effective in stimulating axonal growth than laminin or type IV collagen, either alone or in combination. The results suggest that in addition to laminin and type IV collagen, other components within matrigel may contribute to its ability to support axonal growth. PMID- 9225742 TI - Reduction in p140-TrkA receptor protein within the nucleus basalis and cortex in Alzheimer's disease. AB - It has been hypothesized that the diminished transport of nerve growth factor (NGF) seen within cholinergic basal forebrain (CBF) neurons in Alzheimer's disease (AD) results from a defect in the expression of its high-affinity trkA receptor. The present study used an anti-human trkA-specific monoclonal antibody (mAb 5C3) that recognizes the NGF docking site, combined with quantitative optical densitometry, to evaluate whether expression of the trkA protein is altered within the nucleus basalis and its cortical projection sites in AD. In normal aged humans, trkA immunoreactivity revealed a continuum of positive neurons extending throughout all CBF subfields. In addition, trkA-positive neurons were scattered throughout the olfactory tubercle and striatum. These regions also displayed intense trkA neuropil staining. Although fewer in total number, remaining CBF perikarya in AD displayed a significant decrease in trkA levels relative to aged controls. Biochemical analysis revealed a significant reduction in trkA protein within both the nucleus basalis and the frontal cortex in AD relative to aged controls. In contrast, trkA levels in the caudate nucleus were unaffected. The decrease in trkA protein in conjunction with our recent observations that the message for trkA is reduced within individual CBF neurons in AD supports the concept that defects in the production and/or utilization of the trkA receptor may be a key event mediating degeneration of NGF-responsive CBF neurons in this disease. PMID- 9225743 TI - Prion protein-deficient cells show altered response to oxidative stress due to decreased SOD-1 activity. AB - The cellular function of the prion protein (PrPc), a cell surface glycoprotein expressed in neurones and astrocytes, has not been elucidated. Cell culture experiments reveal that cerebellar cells lacking PrPc are more sensitive to oxidative stress and undergo cell death more readily than wild-type cells. This effect is reversible by treatment with vitamin E. In vivo studies show that the activity of Cu/Zn superoxide dismutase is reduced in Prnp gene-ablated (Prnp0/0) mice. Constitutively high Mn superoxide dismutase activity in these animals may compensate for this loss of responsiveness to oxidative stress. These findings suggest that PrPc may influence the activity of Cu/Zn superoxide dismutase and may be important for cellular resistance to oxidative stress. PMID- 9225744 TI - Descending propriospinal neurons in normal and spinal cord-transected lamprey. AB - The organization and distribution of propriospinal neurons with descending axons were determined via retrograde HRP labeling in normal lamprey and in animals that had behaviorally recovered for various times (4, 8, 16, and 32 weeks) following transection of the rostral spinal cord. In normal animals, descending propriospinal neurons were found in the rostral, middle, and caudal spinal cord. Theoretical analysis indicated that the majority of these neurons had relatively short axons (< 10-15 mm), although a few neurons had relatively long axons (> 30 mm). In spinal cord-transected animals, with increasing recovery times there was a gradual increase in the numbers of labeled propriospinal neurons below the lesion with short-to moderate-length descending axons. The distribution of descending propriospinal neurons and the possible plasticity in this system following spinal cord transection are discussed with regard to activation of spinal motor networks and initiation of locomotor behavior. PMID- 9225745 TI - Expression profile of stress proteins, intermediate filaments, and adhesion molecules in experimentally denervated and reinnervated rat facial muscle. AB - The immunohistochemical profiles of ubiquitin, heat shock protein 70, alpha-B crystallin, desmin, vimentin, neural cell adhesion molecule (N-CAM), and tenascin in rat facial muscle were studied after permanent denervation by transection of the facial plexus on one side and compared with findings after immediate reinnervation by hypoglossal-facial nerve anastomosis subsequent to transection on the contralateral side. Levator labii muscle samples were collected sequentially at 2, 6, 7, 10, 20, and 24 weeks after surgery. Normal levator labii muscle fibers showed physiological expression of desmin and alpha-B-crystallin. Denervated rat facial muscle displayed distinct up-regulation of ubiquitin, alpha B-crystallin, N-CAM, and tenascin. While alpha-B-crystallin and N-CAM decreased in long-standing denervation, tenascin had completely disappeared at 6 weeks. Like-wise, reinnervated muscles displayed enhanced expression of ubiquitin, alpha B-crystallin, N-CAM, tenascin, and, additionally, desmin. Strong expression of desmin and ubiquitin was found up to the 10th week as well as of alpha-B crystallin, N-CAM, and tenascin up to the 7th week of reinnervation. Afterward, expression of stress proteins, intermediate filaments, and adhesion molecules returned to expression profiles of normal controls, indicating that enhancement of these proteins was restricted to the "atrophic and regenerative" states with a decline to physiological levels after successful reinnervation and restoration of muscle fibers. Furthermore part of regeneration from damage seems to resemble reactivated developmental mechanisms by reappearance of developmentally expressed proteins like desmin, N-CAM, and tenascin. PMID- 9225747 TI - Transplantation of mesencephalic cell suspension in dopamine-denervated striatum of the rat. II. Effects on corticostriatal transmission. AB - The present study has been designed to investigate whether intrastriatal implantation of mesencephalic dopamine (DA)-synthetizing neurons into the striatum (ST) of rats whose substantia nigra (SN) was previously destroyed by 6 hydroxydopamine (6-OHDA) restores the pattern of corticostriatal transmission from the medial prelimbic and sensorimotor cortices. In 6-month-old normal animals electrical stimulation of these two functionally unrelated cortices evoked a short latency and brief excitation in 81.6% of neurons recorded in the dorsolateral ST. This percentage decreased significantly (70.6%) in age-matched animals whose dopaminergic nigrostriatal pathway was unilaterally destroyed by 6 OHDA 3 months before recording. However a significant increase in neurons (36.9%) which could be simultaneously activated from the two cortices in comparison to intact rats was noted. In addition the lesion caused a significant decrease in the threshold current required to evoke activation of striatal neurons from the sensorimotor cortex. The increase in the number of striatal neurons responding simultaneously to cortical stimulations demonstrates that destruction of the dopaminergic nigrostriatal pathway causes a loss of the focusing action of DA on corticostriatal transmission. Transplantation of embryonic mesencephalic neurons appears to reestablish this action since the number of convergent responses was significantly decreased in grafted animals (23.5%) in comparison to denervated (36.9%) and sham-grafted (35.1%) animals. Furthermore, the grafts showed a trend to increase current intensities required to evoke activation of striatal cells from both cortices. The action of grafted mesencephalic neurons over prelimbic and sensorimotor cortical inputs to the dorsal ST could be involved in recovery of grafted animals in the correct execution of complex sensorimotor tasks requiring integration of different cortical signals within the ST. PMID- 9225746 TI - Riluzole reduces incidence of abnormal movements but not striatal cell death in a primate model of progressive striatal degeneration. AB - Riluzole has been shown recently to increase life expectancy in patients with amyotrophic lateral sclerosis. A number of experimental studies also suggest that this compound may be a neuroprotectant. We have investigated in baboons whether riluzole would protect striatal neurons from a prolonged 3-nitropropionic acid (3NP) treatment and ameliorate the associated motor symptoms. In animals receiving 3NP and the solvent of riluzole, 12 weeks of high-dose 3NP treatment resulted in the appearance of persistent leg dystonia and significant increases in the incidence of three categories of abnormal movements and in the dyskinesia index in the apomorphine test (0.5 mg/kg i.m.). Quantitative assessment of these behavioral deficits using a video movement analysis system demonstrated a significant decrease in locomotor activity and peak tangential velocity in 3NP treated animals compared to controls. Histological analysis showed the presence of severe, bilateral, striatal lesions, localized in both caudate and putamen. Cotreatment with riluzole (4 mg/kg i.p., twice daily) significantly reduced the dyskinesia index (-35%, P < 0.02) in the apomorphine test. In the quantitative behavioral analysis, riluzole significantly ameliorated the decrease in peak tangential velocity (P < 0.02) but not the decrease in locomotor activity observed after 3NP. Comparative histological analysis of the two groups of treated animals did not demonstrate a clear neuroprotective effect of riluzole. The present study suggests that one potential therapeutic interest for riluzole in neurodegenerative disorders may reside in the reduction of motor symptoms associated with striatal lesions. PMID- 9225748 TI - Capability for reactive gliosis develops prenatally in the diencephalon but not in the cortex of rats. AB - In this study, the glial reactions to stab wounds were investigated on a large population of newborn (P0) and fetal rats, by the immunohistochemical staining of the glial fibrillary acidic protein. The lesions penetrated both the cortex and the diencephalon. The fetuses were lesioned in utero from the 17th embryonic day (E17) and were born on E22 or E23 in the natural way. In the cortex usually no reactive gliosis developed although definitive tissue destructions remained after the lesion. Weak and incomplete glial reactions were observed in a few cases of E20 or P0 lesions only. In the diencephalon, however, the same stabbings provoked massive glial reactions. The timing and the morphology of this reaction were similar to those found in adult animals. At E17 the lesion did not result in reactive gliosis even in the diencephalon. Our study highlights two phenomena: (i) depending on the brain area servere glial reactions can already follow fetal lesions, and (ii) the appearance of the capability for glial reactions may be a stage of the local tissue maturation in every brain area and cannot be considered as a function of brain development in general. Probably, the capability for glial reactions can take place only when certain histogenetic processes (e.g., cell migration, axon growth, apoptosis) have been at least mostly accomplished, but which of the local development events are the determining ones remains to be investigated. PMID- 9225749 TI - Parkinsonism reduces coordination of fingers, wrist, and arm in fine motor control. AB - This experiment investigates movement coordination in Parkinson's disease (PD) subjects. Seventeen PD patients and 12 elderly control subjects performed several handwriting-like tasks on a digitizing writing tablet resting on top of a table in front of the subject. The writing patterns, in increasing order of coordination complexity, were repetitive back-and-forth movements in various orientations, circles and loops in clockwise and counterclockwise directions, and a complex writing pattern. The patterns were analyzed in terms of jerk normalized for duration and size per stroke. In the PD subjects, back-and-forth strokes, involving coordination of fingers and wrist, showed larger normalized jerk than strokes performed using either the wrist or the fingers alone. In the PD patients, wrist flexion (plus radial deviation) showed greater normalized jerk in comparison to wrist extension (plus ulnar deviation). The elderly control subjects showed no such effects as a function of coordination complexity. For both PD and elderly control subjects, looping patterns consisting of circles with a left-to-right forearm movement, did not show a systematic increase of normalized jerk. The same handwriting patterns were then simulated using a biologically inspired neural network model of the basal ganglia thalamocortical relations for a control and a mild PD subject. The network simulation was consistent with the observed experimental results, providing additional support that a reduced capability to coordinate wrist and finger movements may be caused by suboptimal functioning of the basal ganglia in PD. The results suggest that in PD patients fine motor control problems may be caused by a reduced capability to coordinate the fingers and wrist and by reduced control of wrist flexion. PMID- 9225750 TI - Characterization of zinc-induced neuronal death in primary cultures of rat cerebellar granule cells. AB - Although zinc is essential for the activity of numerous biological systems, and zinc deficiency has been associated with various pathologies, this metal can also exert direct neurotoxic action. In primary cultures of rat cerebellar granule neurons, a brief, 15- to 30-min exposure to zinc (100-500 microM) resulted in concentration-dependent delayed neuronal death. The toxicity of zinc depended on the maturity of the neuronal cultures-it was not apparent prior to Day 5 and it reached a plateau at about 9-10 days in vitro. We assayed cell injury by measuring mitochondrial functioning (MTT assay) and cell death with the trypan blue exclusion assay. Apoptosis was assayed by the morphological appearance of cells following fluorescence staining with propidium iodide and by the in situ TUNEL technique. Mitochondrial injury was an early result of zinc treatment. Actinomycin D, an inhibitor of macromolecular synthesis, attenuated delayed cell death. The calcium channel blockers nimodipine and amlodipine reduced both mitochondrial injury and cell death; the blockade of ionotropic glutamate receptors with MK-801 or CNQX was ineffective. These results suggest that calcium channel-blocker-sensitive mitochondrial injury and DNA damage are operative in the protein-synthesis-dependent neurotoxicity of zinc. An imbalance of zinc homeostasis might play a role in the pathophysiology of apoptosis-associated neurodegenerative disorders. PMID- 9225751 TI - 17 beta-estradiol attenuates fimbrial lesion-induced decline of ChAT immunoreactive neurons in the rat medial septum. AB - We investigated the neuroprotective effects of 17 beta-estradiol (E2) on medial septal cholinergic neurons following partial unilateral lesion of the fimbriafornix. Adult female rats were ovariectomized (OVX) and, 5 days later, treated with a single intravenous (iv) injection of an estradiol (E2)-chemical delivery system (E2-CDS) or its vehicle hydroxypropyl-beta-cyclodextrin (HPCD). All rats were subjected to partial unilateral electrolytic fimbrial lesion the following day. At 20 days postlesion, brain slices from treated animals were assessed for choline acetyltransferase (ChAT) by immunohistochemistry. Animals treated with HPCD or E2-CDS showed a 44 or 4% decrease, respectively, in ChAT positive neurons on the lesioned side compared to the nonlesioned side of the medial septum. In a second study using the same lesioning procedure, adult OVX rats received either a subcutaneous E2 pellet implant (n = 6), or, 5 days postovariectomy, a single iv injection of E2-CDS (n = 8) or HPCD (n = 6). Animals treated with HPCD showed a 55% decrease in ChAT-positive neurons on the lesioned side compared to the nonlesioned side of the medial septum. By contrast, rats treated with E2-CDS or E2 pellet had a 14 or 13% decrease, respectively, in ChAT positive neurons. Interestingly, E2 treatment substantially decreased ChAT positive neurons on the nonlesioned side of the medial septum in comparison to control animals. The present study suggests that cholinergic neurons in the medial septum are protected from lesion-induced degeneration by treatments which increase brain E2 levels. Thus, E2 may play a neuroprotective role in the basal forebrain cholinergic system. PMID- 9225752 TI - Comparison of immunohistochemical and functional reinnervation of skin and muscle after peripheral nerve injury. AB - In order to investigate the usefulness of immunohistochemical detection of regenerating axons as a correlate of functional recovery, reinnervation of mouse foot pads, hairy skin, and muscle were studied at several intervals along 3 months after sciatic nerve crush using immunohistochemical markers PGP 9.5 and CGRP. These histological results were compared with functional recovery of sweat glands (SGs), plantar muscles, and pain sensibility. One week after nerve injury all neural functions were abolished in the operated hindpaw of all mice, no CGRP immunoreactive (-ir) fibers were seen in the samples studied, while PGP 9.5 immunofluorescence remained at dim levels within nerve trunks, but disappeared from terminal innervation. The first PGP 9.5- and CGRP-ir regenerating fibers were seen at 15-16 days postoperation (dpo) in dermal nerve trunks of dorsal hairy skin and some days later in dermal trunks of foot pads. Regenerating nerve fibers progressed along the periphery of the dermis reinnervating the different dermal appendages. At 25 dpo all target organs were reinnervated. The first SGs activated by pilocarpine reappeared by 16 dpo and increased in number to 88% of control counts. Nociceptive responses reappeared at 17 dpo and reached 100% of control values. The first PGP immunofluorescence in neuromuscular junctions was seen at 16 dpo, while the first muscle action potentials were recorded at 19 dpo, and the potentials amplitude increased to 66% of controls. Good correlations were found between morphological and functional results of reinnervation. However, the density and distribution of nerve profiles in the tissues studied did not reach normal levels, while neural functions conveyed by small fibers reached levels similar to controls. PMID- 9225753 TI - A subpopulation of reactive astrocytes at the immediate site of cerebral cortical injury. AB - We have identified an early-appearing intermediate filament-associated protein (IFAP-70/280 kDa) in radial glia and their immediate derivatives. This IFAP is absent in the adult CNS. In this study, we examined the reexpression of this early glial differentiation trait in rat reactive astrocytes induced by stab injury of the cerebrum. Double-label immunofluorescence microscopy demonstrated that by 36 h postlesion, IFAP-70/280 kDa was present in a few GFAP-positive astrocytes in the area adjacent to the wound. As the gliotic reaction progressed, the number of IFAP-positive reactive astrocytes increased and by 5-6 days postlesion, IFAP-70/280 kDa was present in most of the hypertrophied astrocytes in tissue immediately adjacent to the wound. By 8 days postlesion, while the number of IFAP-negative reactive astrocytes away from the wound diminished, the IFAP-containing reactive astrocytes close to the wound persisted. Concurrently, they began to change from a stellate form to an elongated shape, with their longitudinal axes radiating from the wound. The immunoreactivity of this IFAP started to diminish at 20 days postlesion, and by 30 days postlesion, it was not observed in the remaining gliotic cells. These results demonstrate that reactive astrocytes induced by stab-wound injury can be divided into two subtypes: persistent IFAP-70/280 kDa-containing cells which are close to the wound in the area of the glial scar and transient IFAP-70/280 kDa-negative cells which are farther from the wound. The reappearance of IFAP-70/280 kDa also suggests that some reactive astrocytes have the capacity to recapitulate early developmental stages. PMID- 9225754 TI - p75 neurotrophin receptor induction and macrophage infiltration in peripheral nerve during experimental diabetic neuropathy: possible relevance on regeneration. AB - In this study we examined the expression of the neurotrophin receptor p75 (p75NTR) and the activation of macrophages in the sciatic nerve of rats at different time points after the induction of diabetes with streptozotocin (STZ). Northern blot and immunocytochemical analysis showed that p75NTR was not detectable in the sciatic nerve by Week 2 after STZ treatment. At this time, single nerve fiber immunostaining using ED1 monoclonal antibody revealed that active macrophages were infiltrating the endoneurium, which had a normal morphological aspect. By Weeks 5 and 15 p75NTR mRNA and protein were induced in the endoneurium of diabetic animals. Immunocytochemical analysis of teased single nerve fibers showed that p75NTR protein was distributed uniformly along isolated fibers with no pathological evidence of axonal degeneration or myelin disruption. At this time, cells of the phagocyte lineage had already disappeared from the nerve. These data show that during experimental diabetic neuropathy, the endoneurial induction of p75NTR is localized along isolated nerve fibers showing no morphological alterations, and in time, follows the recruitment of active macrophages in the nerve, suggesting that these cells, directly or through their products, can influence p75NTR induction. This process might play an important role in STZ diabetic neuropathy, as a response to decreased levels of neurotrophins such as NGF and promoting nerve regeneration in the early phases of the disease. PMID- 9225755 TI - 3-Nitropropionic acid neurotoxicity: visualization by silver staining and implications for use as an animal model of Huntington's disease. AB - The neuronal damage produced by the mitochondrial toxin 3-nitropropionic acid (3NPA) has been suggested to replicate much of the neuropathology seen in Huntington's disease (HD) and therefore might be used in an animal model. We investigated the susceptibility to this toxin of different neuronal populations in addition to the commonly studied caudate putamen by injecting 3NPA into seven different brain regions as well as systemically. After different survival times, rats were intracardially perfused, brain sections were processed with the Gallyas silver technique, and impregnated neurons were mapped with a computerized microscope. Intracerebral administration of 3NPA resulted in a lesion, the center of which was devoid of tissue while the area was surrounded by a halo of Golgi like impregnated neurons. In addition to local damage, rats receiving microinjections into the frontal cortex, caudate putamen, basal forebrain, and amygdala displayed argyrophilic neurons in the thalamus corresponding to the topography of thalamofugal neurons projecting to the individual injection sites. On the other hand, negligible secondary damage was seen after injections into the internal capsule, thalamus, or substantia nigra, implicating that thalamofugal axons are especially vulnerable to the local effect of this toxin. Two and a half days after systemic administration of 3NPA, a diffuse argyrophilic neuronal reaction was seen in the dorsolateral part of the caudate putamen, together with a more regionally selective staining of neurons in different cortical areas and the hippocampus. These morphopathological changes were also accompanied by motor deficits. The affected neurons in the cortical regions were primarily in those layers (V and VI) and areas (medial prefrontal, caudal insular/periphinal, and ventral temporal) that do not project toward the lesioned striatal area; therefore, the cortical pathology may represent another primary site of action of the toxin. Among the affected neurons in the hippocampal complex were pyramidal neurons in the CA1 region as well as various neurons in the CA3 region and dentate hilar area. These studies suggest that a combination of 3NPA administration and a sensitive silver-impregnation method may unravel the potential site of primary neuronal damage in this animal model. Furthermore, these findings may contribute to the understanding of how the disease progresses in HD from the originally affected neuronal population(s) by the recruitment of closely related systems and pathways. PMID- 9225756 TI - Effect of intermittent long-lasting electrical tooth stimulation on pulpal blood flow and immunocompetent cells: a hemodynamic and immunohistochemical study in young rat molars. AB - Release of sensory neuropeptides after stimulation of afferent nerve fibers has previously been shown to induce vasodilation and increased vascular permeability in the dental pulp, a condition recognized as neurogenic inflammation. In the present study a possible role for the sensory neuropeptides in transendothelial migration of immunocompetent cells was investigated. The dental pulp is an isolated tissue densely innervated with sensory fibers containing neuropeptides, and following electrical stimulation of the crown, the effect on pulpal blood flow and immunocompetent cells can be studied in a noninvasive model. A laser Doppler flowmeter was used to measure relative changes in pulpal blood flow during long-lasting intermittent stimulation of innervated and denervated rat first molars. In the innervated teeth, stimulation promptly increased pulpal blood flow by on average 45% at the start of the experiment, whereas almost no blood flow increase was recorded after 4 to 5 h stimulation. Surgical sectioning of the inferior alveolar nerve abolished blood flow increase upon stimulation. After stimulation, a quantitative analysis of CD43+, CD4+, CD11+, and I-A antigen expressing cells was performed, and the effect of stimulation on calcitonin gene related peptide (CGRP)-immunoreactive and substance P (SP)-immunoreactive (IR) nerve fibers was studied. Immunohistochemistry was performed by the avidin-biotin peroxidase method. Stimulation resulted in an almost complete depletion of CGRP- and SP-IR nerve fibers in the first molar pulp, whereas nerve fibers in the gingiva and neighboring teeth were unaffected. A significant increase in the number of CD43+ cells was found in the innervated tooth after stimulation compared to the stimulated denervated (P < 0.01) and unstimulated control (P < 0.05) first molars. For I-A antigen-expressing cells a significant increase (P < 0.05) was found between the innervated stimulated and unstimulated control, but not between the innervated and denervated stimulated first molars. Hence, from the present experiment it is concluded that the pulpal nerves participate in and facilitate transendothelial migration of CD43+ cells during acute neurogenic inflammation. PMID- 9225757 TI - Time course of changes in lactate and free fatty acids after experimental brain injury and relationship to morphologic damage. AB - Regional levels of lactate and free fatty acids (FFA) were measured after lateral fluid percussion (FP) brain injury in rats. At 5 min after injury, tissue concentrations of lactate were elevated in the cortices and hippocampi of both ipsilateral and contralateral hemispheres. Whereas lactate levels had returned to normal by about 20 min after injury in the penumbra and contralateral cortices, their elevation persisted in the ipsilateral injured cortex and hippocampus for 24 h after injury. Increases in the levels of FFA (particularly stearic and arachidonic acids) were observed in the cortices and hippocampi of both ipsilateral and contralateral hemispheres at 5 min after injury; these levels returned to normal in only the penumbra and contralateral cortices by 20 min after injury. Increased amounts of palmitic and oleic acids were also found only in the injured left cortex and ipsilateral hippocampus at 20 min or later after injury. In general, these elevations persisted for as long as 6 to 24 h in the injured cortex and for 2.5 to 24 h after injury in the ipsilateral hippocampus. Histologic studies revealed a similar extent of damage in the cortex between 5 min and 24 h after injury, whereas damage in the CA3 region of the ipsilateral hippocampus increased during that period. These findings suggest a role for lactic acid and FFA, two secondary injury factors, in neuronal cell loss after brain injury. PMID- 9225758 TI - Asymmetrical protection of neostriatal neurons against transient forebrain ischemia by unilateral dopamine depletion. AB - Neurons in the dorsal neostriatum are highly vulnerable to transient cerebral ischemia. It has been suggested that excessive dopamine release during ischemia may play an important role in the pathogenesis of postischemic cell death in the neostriatum. However, it remains controversial whether depletion of dopamine protects neurons in the neostriatum against ischemic insult. In the present study, transient forebrain ischemia was induced using the four-vessel occlusion method. Ischemic depolarization was used as an indication of completed ischemia. Under our experimental conditions, ischemia that produces approximately 21 min ischemic depolarization caused more than 90% of cell death in the dorsolateral neostriatum. Using such ischemia as a standard insult, the effect of dopamine depletion on neostriatal neurons after ischemia was investigated. Dopamine depletion was produced by unilateral injection of 6-OHDA into the substantia nigra. No difference was found between the number of surviving neurons in the left and the right neostriatum after depletion of dopamine on the left side. In contrast, surviving neurons dramatically increased in the right neostriatum after depletion of dopamine on the right side. These results clearly demonstrate an asymmetrical protection of neostriatal neurons against ischemia after dopamine depletion. The mechanisms of this asymmetrical protection may clarify dopamine action on neuronal injury following cerebral ischemia. PMID- 9225759 TI - Gonadal steroid regulation of hamster facial nerve regeneration: effects of dihydrotestosterone and estradiol. AB - We have demonstrated, in a series of experiments, the therapeutic potential of androgens in facial motoneuron regeneration in the adult hamster. Initial work utilized testosterone propionate (TP) as the form of androgen given to adult hamster at the time of facial nerve crush axotomy at its exit from the stylomastoid foramen. TP is capable of being enzymatically converted to estrogen. Thus, the effects of TP on the regenerative properties of facial motoneurons could be due to androgens, estrogens, or both. Recent studies of androgen receptor (AR) mRNA levels suggest that androgens and estrogens work synergistically to regulate AR expression in these motoneurons. In this study, we examined the ability of dihydrotestosterone propionate (DHTP), a nonaromatizable androgen which cannot be converted to estrogen, and estradiol (E2) to alter facial nerve regeneration, using fast axonal transport of radioactively labeled proteins to assess facial nerve regeneration. Adult gonadectomized male hamsters were subjected to right facial nerve crush axotomy, with the left side serving as control, and divided into three groups. One-third of the animals received 1 subcutaneous implant of DHTP, one-third received 1 subcutaneous implant of E2, and the remaining third was sham implanted. Postoperative survival times were 4 and 7 days. As expected, DHTP treatment resulted in an approximately 40% increase in the rate of regeneration, with an associated prolongation in the delay time before sprouting occurred. These effects were slightly greater than previously observed with TP, as might be predicted given the more potent physiological effects observed with DHTP compared to TP. Surprisingly, E2 treatment also resulted in an increase in the rate of regeneration (30%), with minimal effects on the delay time before sprout formation occurred. The results argue for a synergistic role for androgens and estrogens in augmenting peripheral nerve regeneration in the model system used in this study. PMID- 9225760 TI - Differential distribution of immunoreactivity in the adult rat spinal cord revealed by the monoclonal antibody, Py: a light and electron microscopic study. AB - The monoclonal antibody Py has previously been shown to be a useful marker for subpopulations of neurons in the rat brain. However, the distribution of Py immunoreactivity in other regions of the CNS and PNS is not known. Here, we present a light and electron microscopic investigation into the distribution of Py immunoreactivity in the adult rat spinal cord, dorsal root ganglia, and peripheral nerves. Py immunoreactivity was associated with cytoskeletal elements in the cell body and dendrites of large-diameter neurons (particularly motoneurons, Clarke's nucleus neurons, and some dorsal root ganglion cells). Small-diameter neurons of lamina II (substantia gelatinosa) were Py negative. Py immunoreactivity was also detected in some populations of nerve fibers, notably axons located in the corticospinal tract, axons in the region of the white matter bordering the gray matter (presumably propriospinal axons), and also motor axons of the ventral root, but not in peripheral nerve. Dorsal roots were largely unstained. The present observations suggest a possible involvement of the Py antigen in the function or maintenance of the cytoskeleton of some populations of neurons and that the antibody may be a potentially useful tool for studying lesion-induced cytoskeletal alterations, particularly in alpha-motoneurons and Clarke's nucleus neurons. PMID- 9225762 TI - Age-dependent expression of the apamin-sensitive calcium-activated K+ channel in fast and slow rat skeletal muscle. AB - An altered expression of the apamin-sensitive K+ channel from skeletal muscle is apparently implicated in human myotonic dystrophy (MD). We found, in rats, that the expression of this channel depends on age and the type of muscle. This result may be one of the bases of the different susceptibilities of fast and slow muscles to drug-induced myotonia. PMID- 9225761 TI - Myotonic dystrophy: decreased levels of myotonin protein kinase (Mt-PK) leads to apoptosis in muscle cells. AB - The pathogenesis of myotonic dystrophy (DM) and the function of the product of the DM gene, myotonin protein kinase (Mt-PK), and its relationship to the disease are uncertain. To gain insight into the function of Mt-PK we studied the effect of decreasing the levels of Mt-PK in cultured human myoblasts. Myoblasts were transfected with an anti-sense oligonucleotide (ODN) targeted to the translation initiation site of DM mRNA which resulted in about 76% reduction in the levels of Mt-PK protein. A large percentage (about 48 to 90%) of myoblasts transfected with this oligonucleotide (but only about 2 to 23% of myoblasts transfected with a control oligonucleotide) underwent apoptosis within 24 h. To further substantiate these results we delivered a specific antibody to Mt-PK into the myoblast cells using a lipid carrier to inhibit its function and show that this resulted in apoptosis in 57 to 72% of the cells within 24 h. These results suggest that decreased levels of Mt-PK may contribute to muscle pathology in DM by leading to apoptosis of muscle cells. PMID- 9225764 TI - The antioxidant enzymes glutathione peroxidase and catalase increase following traumatic brain injury in the rat. AB - The inflammatory response following mechanical brain injury is characterized by an increase in the cytokine interleukin-1 beta (IL-1 beta) followed by a large elevation in the neurotrophin, nerve growth factor (NGF). The substantial upregulation in NGF observed in our previous studies suggests that it may have functions in addition to that of a target-derived neuronal support mechanism. We hypothesize that NGF is a mediator of oxidative homeostasis, by inducing the production of oxygen-free radical scavengers in brain tissue following injury. We tested this hypothesis by measuring the activity of the antioxidant enzymes, glutathione peroxidase (GSH-Px), and catalase in cortical brain tissue following experimentally induced cortical contusion. We observed a twofold increase in GSH Px and a threefold increase in catalase activities in a time course which reflected the temporal increase in NGF observed following the same cortical contusion model. These findings support the hypothesis and illuminate an important reparative role for NGF following trauma. PMID- 9225763 TI - Flow cytometric analysis of major histocompatibility complex (MHC) class II antigen expression on brain cells from Borna disease virus-infected rats without an intervening in vitro culture step. AB - Borna disease virus (BDV) infects astrocytes in the Lewis rat brain. BDV-infected astrocytes have been shown to express MHC class II in vitro but not in vivo. Using a sensitive fluorescence-activated cytometric technique, we now report the detection of MHC class II on freshly harvested S100-positive cells from BDV infected rat brain, without an intervening in vitro culture step. These data support the hypothesis that astrocytes from BDV-infected rats express MHC class II on their surface and, thus, are potential participants in the encephalitic response to BDV infection. PMID- 9225765 TI - Magnetic resonance imaging of experimental microinfarctions in the rabbit brain. AB - Small hyperintense lesions are frequently found with T2-weighted magnetic resonance imaging (MRI) of the human brain. A significant number of these lesions are probably infarctions. Because there is often a long delay between the microischemic impact and the autopsy, if any, and as the specificity of the MRI is low for detecting ischemic lesions, it is difficult to draw conclusions about the clinicotopographic correlations. This work concerns the usefulness of MRI in detecting experimental microischemic lesions in the rabbit brain about 24 h after the impact. It seems that the sensitivity of T2-weighted MRI in detecting foci of damaged areas in the rabbit brain is good enough to make it useful for evaluating tissue damage when screening for potential neuroprotective drugs and that the experimental model should be useful for developing diagnostically valuable MRI techniques. PMID- 9225766 TI - Detection of Loa loa-specific DNA in blood from occult-infected individuals. AB - Accurate and specific diagnosis of human loiasis is of crucial importance in an endemic area where two-thirds of infected individuals are without circulating microfilariae (occult loiasis). By using the polymerase chain reaction (PCR) and specific primers to the repeat 3 region (15r3) of the gene coding for Loa loa 15 kDa polyprotein antigen, DNA was amplified from total blood lysate of occult infected subjects. A 396-bp DNA fragment was specifically detected. We tested the specificity of this method by qualitative hybridization to PCR products using blood lysates of the following subjects: (1) from Gabon (80 individuals residing in L. loa endemic area where loiasis exists sympatrically with Mansonella perstans); (2) from Togo (12 individuals infected with Onchocerca volvulus and M. perstans); (3) from Tahiti (12 individuals infected with Wuchereria bancrofti); and (4) from Mali (12 individuals infected with O. volvulus and M. perstans). Samples from Gabon included 60 L. loa amicrofilaremics and 20 L. loa occult infected subjects. Qualitative hybridization carried out at 50 degrees C on PCR products, using a 15r3-specific oligonucleotide probe, revealed hybridization with L. loa-infected samples from Gabon and four samples from Togo after 2 days exposure to the film. The positive samples from Togo were characterized by the use of nested PCR. Three nested PCR products have been sequenced. No differences were observed between the three sequences and they are 99.72% identical to L. loa 15r3. None of bancroftian-infected individuals from Tahiti, nor O. volvulus- and M. perstans-infected individuals from Mali reacted after 1 week's exposure (overexposure) to the film. This allows us to conclude first that our 15r3 PCR assay is specific for L. loa and secondly that L. loa infections occur in Togo. The sensitivity of this 15r3 PCR assay was further investigated with occult patients and field-collected amicrofilaremic samples. We found that 19 of the 20 occult-infected individuals were positive on Southern hybridization, whereas 35/60 amicrofilaremics were positive. These results have shown that the sensitivity of this assay in detecting unequivocal, parasitologically proven occult loiasis was 95%, while the specificity with regard to the sympatric M. perstans was 100%. PMID- 9225767 TI - Trypanosoma cruzi: use of herpes simplex virus-thymidine kinase as a negative selectable marker. AB - Trypanosoma cruzi, the protozoan that causes Chagas' disease, was transfected with a fusion gene of hygromycin phosphotransferase and herpes simplex virus thymidine kinase, HyTK. Transfectants selected in hygromycin had thymidine kinase activity, whereas controls did not. In vitro growth of the mammalian life-stage forms, amastigotes and trypomastigotes, was inhibited 98% by the nucleoside analogue ganciclovir (5 micrograms/ml). Growth of the insect-stage form, epimastigotes, was not inhibited by ganciclovir (up to 250 micrograms/ml) or other nucleoside analogues. Intracellular uptake of ganciclovir by epimastigotes was found to be 10-fold less than that by amastigotes. Mice infected with the HyTK-expressing parasites and treated with ganciclovir had a statistically significant reduction of parasitemia by 57%; however, complete eradication of parasites was not achieved. The parasites recovered from the treated mice continued to be susceptible to ganciclovir in vitro. Parasite clones with higher expression of thymidine kinase were more sensitive to ganciclovir, suggesting that greater expression of the thymidine kinase gene may lead to parasites that can be fully eradicated from infected experimental animals. PMID- 9225768 TI - Trypanosoma brucei: lack of cross-resistance to melarsoprol in vitro by cymelarsan-resistant parasites. AB - We have examined cross-resistance between trypanocidal drugs using a well characterised drug-sensitive line, 247, and its cymelarsan-resistant derivative, 247melCyR. The cymelarsan-resistant line was cross-resistant to trimelarsen and melarsen oxide, and partially cross-resistant to two diamidines, pentamidine and berenil (diminazene aceturate). It was cross-resistant to lipid-soluble melarsoprol in vivo but to only a trivial degree in two in vitro assays. The potential role of adenosine transport in arsenical-induced killing of parasites was investigated. Adenosine, adenine, and the diamidines, but not inosine, were able to inhibit killing of drug-sensitive STIB 247 trypanosomes by cymelarsan and melarsen oxide in a concentration-dependent manner. These results are consistent with the view that these arsenical compounds enter trypanosomes via an adenosine specific transporter. Melarsoprol-induced killing of trypanosomes was unaffected, however, by either purine and to only a slight degree by the diamidines. These data suggest that melarsoprol can enter trypanosomes by a route other than through an adenosine transporter and that there may be two mechanisms contributing to arsenical resistance in this drug-resistant line of trypanosomes. PMID- 9225769 TI - Trichinella spiralis: synthesis of type IV and type VI collagen during nurse cell formation. AB - The portion of skeletal muscle fiber (Nurse cell) harboring Trichinella spiralis is surrounded by an acellular capsule susceptible to digestion with collagenase. Antibodies recognized type IV and type VI collagen in the capsule, while the periodic acid Schiff reagent stained the capsule differentially, revealing at least two distinct layers. RNA analysis showed that mRNA specific for type IV and type VI collagen was present in muscle tissue on Days 9 and 15, but not on Day 3, following intracellular infection. In situ hybridization showed that most of the mRNA for both types was within the Nurse cell, and all enlarged Nurse cell nuclei were transcriptionally active for those messages. Synthesis of type IV collagen mRNA was absent by Day 24. In contrast, type VI collagen mRNA was still present at 24 days and 8 months. These results support the hypothesis that T. spiralis, either directly or indirectly, influences the synthesis of these two collagen types throughout its own developmental cycle in the Nurse cell. PMID- 9225770 TI - Homologues of the 24-kDa flagellar Ca(2+)-binding protein gene of Trypanosoma cruzi are present in other members of the Trypanosomatidae family. AB - A full-length cDNA encoding the 24-kDa flagellar Ca(2+)-binding protein (FCaBP) of the Dm28c clone of Trypanosoma cruzi was cloned and characterized. Comparison of the deduced amino acid sequence with those of the FCaBPs of other T. cruzi strains revealed greater than 97% sequence conservation. FCaBP-like genes are found in Trypanosoma conorhini, Trypanosoma freitasi, Trypanosoma lewisi, Herpetomonas megaseliae, Leptomonas seymouri, and Phytomonas serpens, but not in Crithidia deanei, Leishmania amazonensis, or Endotrypanum schaudinni: Among various T. cruzi strains, FCaBP genes are located on chromosomes of different size, although all strains possess multiple FCaBP genes organized as tandemly arranged gene families. Northern and Western blot analyses revealed that FCaBP mRNAs are produced in all organisms possessing FCaBP-hybridizing sequences, indicating that expression of FCaBP or an FCaBP-like protein is common to a number of trypanosomatid species. PMID- 9225771 TI - Transcription of the Leishmania major Hsp70-I gene locus does not proceed through the noncoding region. AB - Primary transcripts in kinetoplastid protozoa are generally assumed to be multicistronic. We have analyzed the transcription in the gene locus which encodes the 70-kDa heat shock protein by using nuclear run-on analysis. We find that RNA synthesis in the Hsp70-I gene locus either is terminated or pauses within the intergenic region approximately 250 nt downstream of the polyadenylation site. We therefore propose a discontinuous mode of transcription in the Hsp70 genes of Leishmania major. PMID- 9225772 TI - Simulium damnosum s.l.: isolation and identification of prophenoloxidase following an infection with Onchocerca spp. using targeted differential display. AB - Phenoloxidase (PO) is the key enzyme for melanin synthesis and plays an important role in the defense and recognition of pathogens in insects and other arthropods. We now report the upregulated transcription of the gene encoding the precursor of PO, prophenoloxidase, in Onchocerca-infected Simulium damnosum s.l., the main vector of human and bovine onchocerciasis in subsaharan Africa. Using homology based generic primers in a polymerase chain reaction-based targeted differential display, the gene itself was identified and partially sequenced. PMID- 9225773 TI - Acanthocheilonema viteae: stage-specific expression of G-protein alpha-subunits. AB - We have previously demonstrated by Western blot analysis that the adult stage of the filarial nematode Acanthocheilonema viteae expresses the alpha-subunits of heterotrimeric G-proteins corresponding to GS and Gq. We now show, using the same technique, that these two alpha-subunits are not detectable in the microfilaria stage of the parasite. Conversely, microfilariae contain Go, an alpha-subunit not expressed by the adult worm. No other G-protein alpha-subunits were found in microfilariae by Western blotting. However, reverse transcriptase-polymerase chain reaction (RT-PCR) with degenerate G-protein oligonucleotide primers, followed by hybridisation analysis, using oligonucleotides specific for individual G-protein alpha-subunits, not only confirmed expression of Go, but also detected Gi1 and G11 alpha-subunits. G-protein expression in infective larvae was also investigated by RT-PCR analysis: this stage of the organism was found to resemble the adult more than the microfilaria but differed from the adult in that GS was absent and Gi3 was present. The significance of these stage specific differences in G-protein expression is discussed with respect to their possible role in parasite development and survival. PMID- 9225774 TI - Plasmodium falciparum: transport of entantiomers of nucleosides into Sendai treated trophozoites. PMID- 9225775 TI - Trypanosoma cruzi: can activity of the rRNA gene promoter be used as a marker for speciation? PMID- 9225776 TI - Plasmodium falciparum: a rapid DNA fingerprinting method using microsatellite sequences within var clusters. PMID- 9225777 TI - Technology assessment, managed healthcare and nuclear medicine. PMID- 9225778 TI - Research radionuclide availability in North America. PMID- 9225779 TI - Clinical validation of the influence of P-glycoprotein on technetium-99m sestamibi uptake in malignant tumors. AB - We prospectively studied 48 patients with either breast cancer (30 patients) or lung cancer (18 patients) to ascertain the relationship between the degree of accumulation of 99mTc-sestamibi and the expression of p-glycoprotein in tumor tissues. METHODS: During initial presentation (37 patients) or post-therapy evaluation (11 patients), the patients underwent contemporaneous 99mTc-sestamibi imaging and biopsy (30 patients) or surgery (18 patients). The interval between surgery/biopsy and imaging varied between 3 and 15 days. All patients had radiologically detectable tumors. Immunohistochemical studies were performed on paraffin sections using a monoclonal antibody, JSB-1, developed against the internal epitope of p-glycoprotein. Tumor-to-background ratios were correlated with the level of p-glycoprotein expression determined by immunohistochemical studies. RESULTS: Our results showed an inverse correlation between the tumor-to background ratios of 99mTc-sestamibi and the density of p-glycoprotein expression in tumor tissues. The values for the tumor-to-background ratios were significantly lower for those tumors expressing p-glycoprotein at high levels than those with scattered and no expression (p < 0.01 and p < 0.001, respectively). CONCLUSION: Although our results warrant further studies at the molecular level using PCR techniques after the extraction of mRNA, our data strongly suggest that 99mTc-sestamibi imaging is useful to noninvasively determine the presence of multidrug resistance in patients with malignant tumors. PMID- 9225780 TI - Monitoring response to therapy with thallium-201 and technetium-99m-sestamibi SPECT in nasopharyngeal carcinoma. AB - This study prospectively assessed the value of 201Tl and 99mTc-sestamibi (MIBI) SPECT in monitoring disease regression/progression as compared with MRI findings in patients with nasopharyngeal carcinoma (NPC) having radiotherapy with or without chemotherapy. METHODS: Eighteen patients (age range 15-78 yr, mean 45 yr) had consecutive SPECT imaging using a dual-head gamma camera after the injection of 111 MBq 201Tl and 555 MBq MIBI before therapy and at 3 mo and 6 mo after completion of therapy. A total of 106 SPECT studies was correlated with contemporaneous MRI studies. Tumor-to-background ratios were obtained on coronal slices. Visually detectable lesions in the region of the nasopharynx and cervical lymph nodes were considered positive for residual disease. The gold standard for the presence of disease was the combination of repeat MRI scans, endoscopic examination and clinical evaluation performed 12-15 mo after completion of therapy. RESULTS: MIBI-SPECT proved superior to both 201Tl SPECT and MRI after 3 or 6 mo follow-up in predicting complete response. Accuracy rates in the detection of residual disease in the nasopharynx are 39%, 72% and 89% for MRI, 201Tl and MIBI, respectively, for the 3-mo evaluation; 71%, 71% and 94% for MRI, 201Tl and MIBI, respectively, for the 6-mo evaluation. CONCLUSION: MIBI SPECT could be used as a screening test in predicting response to therapy in patients with NPC. PMID- 9225781 TI - Technetium-99m-tetrofosmin SPECT imaging of lung masses: a negative study. AB - Technetium-99m-tetrofosmin has emerged as a new radiopharmaceutical for myocardial imaging, in competition with 201Tl and 99mTc-MIBI. In this study, 99mTc-tetrofosmin was evaluated for its ability to detect malignant and benign lesions from single solid lung masses. METHODS: Forty-nine patients with a single solid lung mass based on chest radiograph findings received 99mTc-tetrofosmin SPECT of the chest to evaluate the value of 99mTc-tetrofosmin SPECT for detecting malignant and benign lesions. RESULTS: Only 61% of the lung malignancies were detected by 99mTc-tetrofosmin SPECT of the chest, including 53% of epidermoid carcinoma (ca), 67% of adeno ca, 75% of small-cell ca, 0% of undifferentiated large-cell ca and 100% of other lung malignancies. In addition, 50% of the benign lesions were detected by chest 99mTc-tetrofosmin SPECT. The probability of tetrofosmin uptake in the mass was not related to mass size. The diagnostic sensitivity, specificity and accuracy were 61%, 50% and 59%, respectively, for differentiating malignant and benign lesions when diagnosing a single solid lung mass. CONCLUSION: Technetium-99m-tetrofosmin SPECT of the chest is of little or no value for the detection of lung ca from single solid lung masses. PMID- 9225782 TI - Radionuclide-guided stereotactic prebiopsy localization of nonpalpable breast lesions with normal mammograms. AB - Scintimammography with 99mTc-sestamibi can be used as a complementary technique to improve the mammogram's sensitivity and specificity for detection of breast carcinoma. We have observed in some patients focal areas of increased 99mTc sestamibi uptake with no corresponding abnormalities on physical examination or mammogram. A phantom device and a special needle were designed to stereotactically localize these lesions before biopsy. METHODS: After intravenous injection of 30 mCI (1110 MBq) of 99m Tc-sestamibi, a prone lateral image of the abnormal breast was obtained. With the patient in the prone position, the breast was compressed with two fenestrated plates in the prone position. The x and y coordinates of the abnormal hot spot of the breast were determined. The z coordinate of focal 99mTc-sestamibi uptake was determined by advancing a localizer needle through a selected predetermined hole of the fenestrated plate using real-time visualization on the persistence monitor. The tip of the opturator inside the needle is welded with 57Co to determine the depth of the hot spot in the breast. RESULTS: Three women, all of whom had normal mammogram and breast physical examinations, were studied using 99mTc-sestamibi prone breast imaging. Pre-excisional biopsy needle localization of abnormal focal uptake was performed. Two women demonstrated infiltrative ductal carcinoma, and the third had proliferative fibrocystic disease of the breast. CONCLUSION: Our initial experience demonstrates that nuclear medicine guided stereotactic needle biopsy of the breast in patients with positive scintimammograms is technically feasible. In the future, this technology will enable us to detect breast carcinoma in the absence of clear-cut clinical and mammographic findings. PMID- 9225783 TI - Frontal sinus mucocele mimicking a metastasis of papillary thyroid carcinoma. AB - Radioiodine scans are highly specific for detecting metastases of well differentiated thyroid carcinomas. However, false-positive 131I whole-body scans may occur as illustrated in the following case report. In a 53-yr-old patient, abnormal 131I uptake was found in the right frontal skull 4 wk after total thyroidectomy and radioiodine therapy for papillary thyroid cancer. Bone scans and planar x-rays of the skull were unremarkable and the serum thyroglobulin level was within normal limits. X-ray CT revealed a small nodule in the right frontal sinus corresponding to the pathological focus of 131I uptake. Surgical removal and histopathological examination of this lesion yielded a mucocele, a slow-growing lesion of the paranasal sinuses accumulating mucous material. The postoperative 131I scan was unremarkable. The possibility of a false-positive finding on radioiodine scans should be considered, particularly when the serum thyroglobulin level is not elevated. PMID- 9225784 TI - Acute liver necrosis induced by iodine-131-MIBG in the treatment of metastatic carcinoid tumors. AB - Iodine-131-metaiodobenzylguanidine (MIBG) is used in the treatment of carcinoid tumors. Temporary palliation with complete subjective symptomatic response has been reported in these patients. This treatment is usually well tolerated and side-effects are generally limited to nausea, mild hepatic toxicity with spontaneous recovery and temporary myelosuppression. Our case report shows that repeated treatment with [131I]MIBG in a patient with extensive carcinoid liver metastasis may cause severe hepatic toxicity leading to death. Factors such as concomitant use of 5-fluorouracil and the progressive nature of the disease may have contributed to this event. PMID- 9225785 TI - FDG hypermetabolism associated with inflammatory necrotic changes following radiation of meningioma. AB - PET with 18F-fluoro-2-deoxy-D-glucose (FDG) is currently the noninvasive gold standard for distinguishing brain tumor recurrence from radiation necrosis. We present a case report that appears to contradict this doctrine. The patient had a history of atypical meningioma and was treated with surgical resection and postoperative proton-beam radiation therapy. Approximately 16 mo after completion of therapy, MRI demonstrated two new regions of enhancement, and an FDG-PET study was performed to further characterize these lesions. FDG-PET demonstrated an area of intense hypermetabolism, and wide surgical resection was performed. Histological examination of the surgical specimen revealed reactive changes and areas of necrosis. There was no evidence of either recurrent or radiation-induced tumor. PMID- 9225786 TI - Comparative PET imaging of experimental tumors with bromine-76-labeled antibodies, fluorine-18-fluorodeoxyglucose and carbon-11-methionine. AB - The potential of a 76Br-labeled anti-carcinoembryonic antigen monoclonal antibody (MAb), 38S1, as tumor-imaging agent for PET was investigated in a comparative experimental study with [18F]fluorodeoxyglucose ([18F]FDG) and L-[methyl 11C]methionine ([11C]Met). METHODS: The three radiotracers were administered to nude rats carrying subcutaneous xenografts or liver metastases from a human colonic carcinoma. Tracer biodistribution was evaluated by PET imaging and radioactivity measurement of dissected tissues and also by whole-body autoradiography for subcutaneous xenografts. RESULTS: For PET imaging of subcutaneous tumors, 76Br-38S1 proved superior to the other radiotracers. Tumor to-tissue ratios were, except for the tumor-to-blood ratio, generally higher for 76Br-labeled MAb than for [18F]FDG and [11C]Met. Liver metastases were imaged with PET using both 76Br-38S1 and [18F]FDG, and the metastases-to-liver ratios of dissected samples were not significantly different for the two radiotracers. CONCLUSION: The tumor-imaging capacity of 76Br-labeled MAb 38S1 was superior to [18F]FDG and [11C]Met in the subcutaneous tumor model, whereas 76Br-38S1 and [18F]FDG were equally successful for the identification of liver metastases. PMID- 9225787 TI - In vitro and in vivo detection of functional somatostatin receptors in canine insulinomas. AB - Ten dogs with hypoglycemia due to insulinomas were studied to assess the expression of somatostatin receptors (SSTRs) in canine insulinomas and its potential diagnostic value. METHODS: The response of circulating glucose and insulin concentrations to the subcutaneous administration of a somatostatin analog, octreotide, was measured. SSTRs were visualized in vitro by autoradiography. [Iodine-125-Tyr3]-octreotide and [125I-Tyr11]-somatostatin-14 (SRIF-14) were used as radioligands. SPECT was performed 6 hr after the injection of [111In-DTPA-D-Phe1]-octreotide. RESULTS: After subcutaneous injection of 50 micrograms octreotide, plasma glucose concentration rose from 2.3 +/- 0.2 mmol/liter to 3.2 +/- 0.3 mmol/liter at 3.5 hr (p < 0.05) and plasma insulin concentration decreased from 451 +/- 135 pmol/liter to a nadir of 249 +/- 115 pmol/liter at 30 min (p < 0.05). In vitro autoradiography revealed that all primary insulinomas and their metastases had specific SSTRs for both [125I-Tyr3] octreotide and [126I-Tyr11]-SRIF-14. Scatchard analysis of SSTR binding in the tumor tissue of one dog revealed high-affinity binding sites for [125I-Tyr3] octreotide (dissociation constant (Kd) 1.7 nM, maximum binding capacity (Bmax) 499 fmol/mg membrane protein). The primary tumor and/or metastases in five of six dogs could be visualized and localized by SPECT with [111In-DTPA-D-Phe1] octreotide. In the remaining dog, multiple metastases (< 3 mm) were found in the liver at necropsy, apparently too small to be visualized by SPECT. CONCLUSION: The in vitro autoradiography and ligand binding studies indicate that canine insulinomas express one type of SSTR. This is in contrast with findings in humans where, on the basis of ligand binding studies, different subtypes of SSTRs have been identified. The uniformity of SSTRs, their high frequency of expression and the high incidence of metastatic disease make canine insulinomas very suitable for investigation of the value of SRIF analogs in the diagnosis and treatment of metastasized endocrine pancreatic tumors. PMID- 9225788 TI - Synthesis, biodistribution and imaging properties of indium-111-DTPA-paclitaxel in mice bearing mammary tumors. AB - Paclitaxel, an antineoplastic agent that stabilizes microtubules and arrests cells in the G2/M cell cycle phase, has shown activity against many common cancers, including ovarian and breast tumors. In order to evaluate the potential value of radiolabeled paclitaxel as an imaging tool in tumors, we synthesized 111In-DEPA-paclitaxel and investigated its biodistribution and gamma scintigraphic imaging properties. METHODS: Mice bearing a paclitaxel-responsive mammary tumor (MCA-4) were used. DTPA-paclitaxel was labeled with 111In with a radiochemical yield of 84% and radiochemical purity of 90%. Each mouse received 5 microCi of radiotracers intravenously for biodistribution studies and 100 microCi for gamma scintigraphic studies. Indium-111-DTPA was used as a control. RESULTS: In tumor-bearing mice, 111In-DTPA was characterized by rapid clearance from the plasma with negligible retention in the tumor, the liver and other body parts. In contrast, 111In-DTPA-paclitaxel exhibited a pharmacological profile resembling that of paclitaxel. Furthermore, a significant uptake of 111In-DTPA-paclitaxel was observed in the tumor. The tumor-to-muscle ratios were 2.64, 3.16 and 6.94 at 30 min, 2 hr and 24 hr, respectively, although absolute uptake in the tumor decreased from 1.95% (injected dose/g) at 30 min to 0.21% at 24 hr after injection. The tumor-to-blood ratio reached 50 at 24 hr after injection. Gamma scintigraphy and autoradiographic studies clearly showed the retention of radiolabeled paclitaxel in the tumor 24 hr after injection. CONCLUSION: These studies suggest that 111In-DTPA-paclitaxel may be clinically useful in studying the uptake of paclitaxel in solid tumors. PMID- 9225789 TI - Multitracer studies during gene therapy of hepatoma cells with herpes simplex virus thymidine kinase and ganciclovir. AB - Using different tracers of tumor metabolism, the application of PET for monitoring gene therapy with the suicide gene herpes simplex virus thymidine kinase (HSVtk) is investigated in this in vitro study. METHODS: Morris hepatoma cells were transfected with a retroviral vector bearing the HSVtk gene, and different clones were established by selection with the neomycin analog G418. Thereafter, uptake measurements using fluorodeoxyglucose (FDG), 3-O methylglucose, aminoisobutyric acid and methionine were performed in a thymidine kinase (TK)-expressing cell line and in control cells bearing the empty vector in the presence of different concentrations of ganciclovir (GCV). These experiments were done up to 48 hr after the onset of therapy. The values were expressed as Bq/well or as Bq/10(5) cells. RESULTS: During GCV treatment therapy, a decrease of the uptake/well was measured for all tracers in the TK-expressing cell line. After normalization to the viable cell number, the uptake for FDG and 3-O methylglucose increases up to 195% after 24 hr incubation with GCV. A high pressure liquid chromatography analysis revealed a decline of the FDG-6-phosphate fraction after 48 hr incubation with GCV. Consequently, a normalization of FDG uptake was observed after this incubation period, whereas the 3-O-methylglucose uptake was still increased. Experiments performed with different amounts of TK expressing cells and control cells showed that these effects are dependent on the percentage of TK-expressing cells. The aminoisobutyric acid uptake decreases to 47%, while the methionine uptake decreases in the acid-insoluble fraction (to 17%) and increases in the acid-soluble fraction (to 150%). CONCLUSION: These data indicate that combinations of the PET tracers used in these experiments may be applied for monitoring gene therapy with HSVtk. The increase in FDG and 3-O methylglucose uptake in vitro is interpreted as stress reaction of the tumor cells. However, an uncoupling of transport and phosphorylation was observed after 48 hr incubation. The amino acid uptake experiments point to an inhibition of protein synthesis as well as of the neutral amino acid transport. PMID- 9225790 TI - Preclinical studies of indium-111-labeled IgM: a human monoclonal antibody for infection imaging. AB - Indium-111-labeled plasma proteins, such as albumin, transferrin and IgG, have been proven useful to image infection. We reported previously that 111In-labeled human monoclonal antibody, IgM 16.88 (In-IgM) also would localize at the site of infection. However, the kinetics of blood clearance, distribution and infection uptake have not been investigated. We compared the kinetics of distribution and infection uptake of In-IgM 16.88 with that of in-polyclonal IgG in rats with focal infection. METHODS: Both IgM 16.88 and polyclonal IgG were labeled with 111In using a bifunctional chelating agent, LiLo. The labeling efficiency was > 95%. Focal infection was induced in rats by an intramuscular injection of E. Coli in the right thigh. In-IgM (30-40 microCi) was injected into five groups of rats (five rats/group). The rats were killed at 4, 8, 16, 24 and 36 hr. The percent injected dose (%ID) in blood, infection muscle, control muscle, liver, spleen and kidney were determined. Similar studies were performed with In-IgG. RESULTS: The In-IgM activity in blood at 4 hr postinjection was 27% which decreased to 2% by 36 hr. In contrast, the In-IgG blood activity was 40% at 4 hr and 20% at 36 hr. The infection/ muscle (I/M) ratios are higher with In-IgM at all time points postinjection compared to that of In-IgG. At 24 hr, the I/M ratio was 22 compared to 9 with In-IgG. At the same time point, the infection/ blood (I/B) ratio with In-IgM was 2.7 compared to only 0.8 with that of In-IgG. In-IgM was taken up mostly by the liver compared to diffuse abdominal uptake of IgG. CONCLUSION: These result indicate that In-IgM produces higher lesion to background ratio when compared to In-IgG and, therefore, is potentially useful to image infection in patients. PMID- 9225791 TI - Three-dimensional dosimetry for intralesional radionuclide therapy using mathematical modeling and multimodality imaging. AB - A method of dosimetry is described that quantifies the three-dimensional absorbed dose distribution resulting from an intralesional administration of a radiolabeled monoclonal antibody, allowing for both spatial and temporal heterogeneity of distribution of the radionuclide and without the need for a calibration scan. METHODS: A mathematical model was developed to describe the distribution of activity as a function of time resulting from infusion at a single point within the solid component of a tumor. The parameters required for this model are either known directly or may be obtained from SPECT image data registered to computed tomography. Convolution of this distribution with a point source dose kernel enabled the three-dimensional absorbed-dose distribution to be obtained. RESULTS: This method was applied to a set of patient data acquired in the course of a clinical study performed at our center, and dose profiles and dose-volume histograms were produced. It was shown that the three-dimensional distribution of dose was significantly nonuniform. CONCLUSION: Initial results suggest that this method offers a means of determining the absorbed dose distribution within a tumor resulting from intralesional infusion. This method extends the Medical Internal Radiation Dose computation, which, in these circumstances, would make erroneous assumptions. Furthermore, it will enable individual patient treatment planning and optimization of the parameters that are within the clinician's control. PMID- 9225792 TI - Quantification of systolic count increase in technetium-99m-MIBI gated myocardial SPECT. AB - This study was performed to clarify the validity of quantification of myocardial wall thickening by the count increase method using electrocardiography (ECG) gated SPECT. METHODS: We performed a phantom study to examine the quantification of this method and to clarify the relationship between the changes of relative counts and objective size (such as myocardial wall) under various conditions. In addition, in volunteers, the percent count increase (%CI) was analyzed in left ventricular segments based on circumferential profile curve analysis by ECG-gated SPECT with 99mTc-MIBI (methoxyisobutyl-isonitrile), and it was compared with the regional systolic wall thickness (%Th) assessed by echocardiography during low dose dobutamine infusion. RESULTS: In our phantom study, the relative count changes were correlated linearly with the object size only within less than 20 mm. Recovery coefficient curves were influenced by acquisition parameters such as type of collimator, diameter of camera rotation, counts and photon scattering. In ECG-gated SPECT, the %CI value was increased gradually at each stage after dobutamine infusion, in relation to the increase of the %Th seen on echocardiography, although there are significant large deviations between these two parameters. CONCLUSIONS: In this study, quantitative analysis based on the %CI in ECG-gated SPECT may underestimate regional wall thickening. These data should be considered in the evaluation of the %CI as an index of myocardial function. PMID- 9225793 TI - Significance of late redistribution thallium-201 imaging after rest injection for detection of viable myocardium. AB - The aim of this study was to determine whether late redistribution imaging after rest injection of 201Tl would provide further information on myocardial viability over conventional rest-early redistribution 201Tl imaging. METHODS: Twenty-nine patients with coronary artery disease and left ventricular dysfunction underwent rest, early (3-4 hr) and late (20-24 hr) redistribution 201Tl and gated blood pool studies. In 14 patients with successful revascularization, gated blood pool study was repeated after the coronary intervention. RESULTS: Nine of 29 patients showed early redistribution, and six additional patients showed further redistribution on the late images. Of 136 segments with initial 201Tl defects, 18 showed early redistribution, and 10 showed late redistribution. When a threshold of 60% of peak activity was used as an index of myocardial viability, only a small fraction (3%) of the initial 201Tl defects were additionally considered viable by the late images. In 14 patients who underwent revascularization, the positive (69%) and negative (87%) predictive values of the early redistribution images for functional recovery were similar to those obtained by the late images (68% and 86%, respectively). CONCLUSION: Although late redistribution after rest injection of 201Tl occasionally occurs, most of the clinically relevant information on myocardial viability may be obtained by conventional rest-early redistribution 201Tl imaging when the defect severity is considered an index of tissue viability. PMID- 9225795 TI - ACE inhibition reduces cardiac iodine-123-MIBG release in heart failure. AB - Radioiodinated metaiodobenzylguanidine (123I-MIBG), an analog of norepinephrine, has been used to assess cardiac sympathetic nerve activity. Decreased myocardial accumulation and enhanced washout of 123I-MIBG have been reported in patients with congestive heart failure (CHF). The purpose of this study was to determine whether angiotensin converting enzyme (ACE) inhibition reduced 123I-MIBG release and improved cardiac 123I-MIBG accumulation in patients with CHF. METHODS: Twenty nine patients receiving conventional treatment for CHF, New York Heart Association (NYHA) functional class 2-3, were studied. Nineteen patients received additional treatment with enalapril, an ACE inhibitor, and 10 patients who were treated with conventional therapy alone were defined as a control group. Iodine 123-MIBG imaging and echocardiography were performed on all patients before treatment and repeated after 9.1 +/- 3.0 mo of treatment. Images were obtained 30 min and 4 hr after injection of 123I-MIBG, and a heart to mediastinum (H/M) ratio was defined to quantify cardiac 123I-MIBG uptake as a fraction of the mean counts per pixel in the heart divided by those in the mediastinum. The washout rate of 123I-MIBG from the heart was calculated as follows: (early counts - delayed counts)/early counts x 100(%). RESULTS: In patients with enalapril group, the H/M ratio of 123I-MIBG was increased after treatment (early image: 1.60 +/- 0.22 vs. 1.73 +/- 0.28, p < 0.05, delayed image: 1.63 +/- 0.28 vs. 1.82 +/- 0.33, p < 0.01). The washout rate of 123I-MIBG was reduced from 38% +/- 11% to 30% +/- 12% after treatment (p < 0.01). However in the conventional therapy group, the H/M ratios in the early and delayed images (early image: 1.58 +/- 0.31 vs. 1.52 +/- 0.23, delayed image: 1.49 +/- 0.27 vs. 1.49 +/- 0.25) and the washout rate (34% +/- 8% vs. 33% +/- 7%) remained unchanged after treatment. In patients with an increased H/M ratio of enalapril group (n = 13), a left ventricular ejection fraction increased from 48% +/- 12% to 55% +/- 9% (p < 0.01) after treatment. CONCLUSION: ACE inhibition reduces cardiac 123I-MIBG release and thus lowers cardiac sympathetic nerve activity. Iodine-123-MIBG may be helpful in evaluating the therapeutic effects of ACE inhibition on the cardiac sympathetic nervous system in patients with CHF. PMID- 9225794 TI - Influence of blood substrate levels on myocardial kinetics of iodine-123-BMIPP. AB - To evaluate the influence of blood substrate levels on myocardial uptake of 123I labeled beta-methyl-iodophenyl-pentadecanoic acid (BMIPP), we examined the correlation between myocardial BMIPP uptake and blood levels of free fatty acid (FFA), glucose, insulin, triglyceride and total cholesterol. METHODS: In 180 patients, venous blood samples were obtained, and the early and late myocardial uptakes (MU15 and MU150) were determined on planar images at 15 and 150 min after injection at rest, respectively, and the clearance rate of BMIPP from the myocardium was calculated. Dynamic SPECT with BMIPP, PET with [18F]fluoro deoxyglucose and determination of myocardial carnitine contents were performed in 15, 1 and 3 patients, respectively. RESULTS: In the 180 patients, MU15 correlated with blood insulin (r = 0.22, p = 0.005) and FFA (r = -0.19, p = 0.02) levels, whereas MU150 did not correlate with blood levels of any variables that were measured (p > 0.05). The clearance rate correlated with blood insulin (r = 0.28, p < 0.001), glucose (r = 0.17, p = 0.03) and FFA (r = -0.40; p < 0.001) levels. The correlations were, however, weak, and five patients (2.8%) with no myocardial BMIPP uptake, all of whom had anterior myocardial infaction, had no characteristics regarding the blood substrate levels. Although dynamic SPECT demonstrated rapid myocardial extraction of BMIPP in 13 patients with myocardial BMIPP uptake, it demonstrated no myocardial BMIPP extraction in two patients with no myocardial BMIPP uptake. One of the five patients with no myocardial BMIPP uptake showed increased myocardial [18F]fluorodeoxyglucose uptake and decreased myocardial carnitine content. CONCLUSION: The influence of blood substrate levels on myocardial BMIPP uptake is not very significant, although high serum FFA levels may be associated with slow clearance of BMIPP from the myocardium. The complete absence of myocardial BMIPP uptake is not rare and may not be associated with changes in blood substrate levels or early back diffusion of BMIPP. PMID- 9225796 TI - Adenosine coronary vasodilation in coronary artery disease: technetium-99m tetrofosmin myocardial tomography versus echocardiography. AB - This study compared the results of adenosine 99mTc-tetrofosmin cardiac tomography with those of adenosine echocardiography in identifying patients with coronary artery disease (CAD) and in localizing individual stenosed, coronary vessels. METHODS: Twenty-six consecutive patients with suspected or known CAD had simultaneous adenosine (140 micrograms/Kg/min intravenously) 99mTc-tetrofosmin tomography and two-dimensional echocardiography. All patients had coronary angiography within 4 wk from imaging studies. Regional 99mTc-tetrofosmin activity was quantitatively measured in 78 coronary vascular territories and echocardiographic left ventricular function was assessed in corresponding regions. RESULTS: At coronary angiography one patient had normal coronary vessels, 12 patients one-vessel and 13 had multivessel disease (> or = 50% luminal stenosis). Among the 25 patients with CAD, 22 showed perfusion defects at adenosine 99mTc-tetrofosmin tomography (sensitivity 88%) and 17 had abnormal echocardiographic study (sensitivity 68%, p < 0.05 versus 99mTc-tetrofosmin). Agreement for the identification of patients with CAD between adenosine 99mTc tetrofosmin tomography and echocardiography was observed in 21 (81%) of the total 26 patients, with a kappa value of 0.45. Overall sensitivity, specificity and diagnostic accuracy for detection of individual stenosed vessels were 79%, 88% and 83% for 99mTc tetrofosmin and 57%, 68% and 61% (all p < 0.05 versus 99mTc tetrofosmin) for echocardiography. Concordance between adenosine 99mTc tetrofosmin tomography and echocardiography in the detection of individual stenosed coronary vessels was observed in 57 (73%) of the 78 vascular territories, with a kappa value of 0.36. CONCLUSION: Adenosine-induced coronary vasodilation associated with quantitative 99mTc-tetrofosmin tomography is more accurate than adenosine echocardiography in identifying patients with CAD and in detecting individual stenosed coronary vessels. PMID- 9225797 TI - Quantitative assessment of cerebral blood flow in patients with Alzheimer's disease by SPECT. AB - This study evaluated an automated analysis of SPECT brain imaging in patients with Alzheimer's disease (AD). METHODS: Patients [n = 81; mean age, 69.9 +/- 10.6 yr (mean +/- s.d.)] with a clinical diagnosis of probable AD (NINCDS-Alzheimer's Disease and Related Disorders Association criteria) underwent 99mTc-ethyl cysteine dimer SPECT imaging. After imaging registration and data extraction using three-dimensional stereotactic surface projections, a pixel-wise comparison of ethyl cysteine dimer uptake was performed using a reference database of 10 cognitive intact controls of comparable age. RESULTS: When individual cases were compared to the normal database, temporo-parietal regional cerebral blood flow (rCBF) abnormalities across different levels of dementia severity were clearly depicted on pixel-wise Z-score images. The rCBF reduction in cortical association areas showed a significant correlation with an overall level of cognitive decline, as assessed by the Mini Mental State Examination and by the cognitive section of the Cambridge Mental Disorders of the Elderly Examination. In addition, there were significant region-specific correlations between left temporo perfusion deficit and language performance and between right parietal rCBF reduction and praxis. CONCLUSION: These results indicate that this observer independent analysis of SPECT data enables objective and semiquantitative assessment of the magnitude and extent of cortical perfusion abnormalities in patients with AD. PMID- 9225798 TI - Regional cerebral blood flow measurement with iodine-123-IMP autoradiography: normal values, reproducibility and sensitivity to hypoperfusion. AB - We recently proposed a simplified technique for measuring regional cerebral blood flow (rCBF) using the [123I]N-isopropyl-p-iodoamphetamine (IMP) autoradiographic (ARG) method with SPECT (the IMP-ARG method). We examined normal values of rCBF and the reproducibility and sensitivity to hypoperfusion in stroke patients using this method. METHODS: By using a standard arterial input, a single static scan, a fixed distribution volume (Vd) and one-point arterial blood sampling, we measured rCBF in 39 normal volunteers (19 men and 20 women; mean ages 61 +/- 11 yr for the men and 60 +/- 12 yr for the women). Eighteen neurologically stable patients with prior stroke (mean age = 65 +/- 11 yr) were studied twice at a mean interval of 97 days. In 16 patients (7 men and 9 women, mean age = 63 +/- 5 yr) with subarachnoid hemorrhage, rCBF was measured 1-2 wk after onset. Cerebral vasospasm was evaluated by repeated angiography. The mean rCBF in the vasospastic area was compared with that in a nonvasospastic area. RESULTS: The mean rCBFs of the cerebral cortex and centrum semiovale in the volunteers were 33.0 +/- 5.1 ml/100 g/min and 25.0 +/- 4.5 ml/100 g/min, respectively. There was no age-dependent change in rCBF, but the women showed significantly higher cortical rCBF than the men (p < 0.05). In the stroke patients, the whole-brain CBF values showed high reproducibility, with high correlations between those obtained at the first and second studies (y = -3.5 + 1.03x; r = 0.90; p < 0.001). In the subarachnoid hemorrhage patients, the vasospastic area showed significantly lower rCBF than the normal cortical rCBF (p < 0.01) and the nonvasospastic area (p < 0.01). Brain regions with rCBF levels below 20 ml/100 g/min showed infarction on the follow-up CT scan. CONCLUSION: The IMP-ARG method is reproducible, sensitive to hypoperfusion and feasible for the quantitative evaluation of rCBF in routine clinical practice. PMID- 9225799 TI - Intra-individual differences between technetium-99m-HMPAO and technetium-99m-ECD in the normal medial temporal lobe. AB - Regional distributions of 99mTc-hexamethyl propyleneamine oxime (99mTc-HMPAO) and 99mTc-ethyl cysteinate dimer (99mTc-ECD) were compared in the normal brain. METHODS: Six paid, healthy volunteers (mean age 26 yr) had high-resolution neuroperfusion SPECT using both 99mTc-HMPAO and 99mTc-ECD on separate days. RESULTS: Regional distribution of the two tracers differed. Technetium-99m-HMPAO accumulated more in the thalamus, frontal lobe, temporal lobe and cerebellum than 99mTc-ECD, which accumulated more in the occipital and parietal lobes. There was a considerable difference in the accumulation of the two tracers in the medial temporal lobe. The percent accumulations of 99mTc-HMPAO and 99mTc-ECD in the medial temporal lobe compared with the mean global cerebral cortical accumulation were 93.9% +/- 2.4% and 83.1% +/- 4.1% (mean +/- s.d.), respectively. CONCLUSION: The results suggest that 99mTc-HMPAO and 99mTc-ECD require specific and separate criteria for diagnosing temporal lobe pathologies, such as dementia and temporal lobe epilepsy. PMID- 9225800 TI - Technetium-99m-HMPAO brain SPECT in systemic lupus erythematosus with CNS involvement. AB - Functional brain SPECT is playing an increasingly important role in evaluating CNS conditions in patients with systemic lupus erythematosus (SLE). However, SPECT findings varied in different studies because of their small population. Furthermore, earlier researchers, being restricted by the resolution of the camera, might not have been able to evaluate deep-seated nuclei such as the basal ganglia and thalamus. In this study, we describe the different patterns of SPECT findings in SLE patients with CNS involvement. METHODS: Seventy-two SLE patients (aged 14-67 yr; mean 33.2 yr) were divided into three groups: Group 1 with definite neuro-psychiatric disorder (including stroke, seizures and psychosis); Group 2 with minor neuropsychiatric disorders (headache, dizziness and recent memory impairment); and Group 3 without any neuropsychiatric symptoms or signs. Ninety minutes after injection of 1110 MBq 99mTc-HMPAO, brain SPECT was performed using a dual-head camera and fan-beam collimator. In addition, MRI and an electroencephalography (EEG) were also performed. RESULTS: SPECT findings were normal in 87% of the Group 3 patients and abnormal in all Group 1 patients; 84.6% of the Group 2 patients had abnormal SPECT findings. The parietal, frontal and temporal lobes were the most common areas of CNS involvement. Parietal lobes were involved in 95.6% of Group 1 patients and 80.7% in Group 2 patients. Frontal lobes were involved in 56.5% of Group 1 patients and 65.3% of Group 2 patients. Temporal lobes were involved in 56.5% of Group 1 patients and 46.1% of Group 2 patients. The basal ganglion was involved in about 30% of Group 1 patients and 11.5% of Group 2 patients, while the thalamus and cerebellum were less involved in neuropsychiatric SLE. MR images showed less sensitivity in the detection of CNS involvement than the SPECT and were normal in 27.3% of patients with definite neuropsychiatric disorders. The EEG and anticardiolipin antibody did not correlate well to the clinical diagnosis. CONCLUSION: HMPAO brain SPECT had the best correlation with the clinical diagnosis and may provide additional and objective information on SLE patients with potential CNS involvement. PMID- 9225801 TI - Normal brain perfusion pattern of technetium-99m-ethylcysteinate dimer in children. AB - The purpose of this study was to assess the normal perfusion pattern of the pediatric brain with 99mTc-ethylcysteinate dimer (99mTc-ECD). METHODS: Tomographic imaging was performed with a dedicated system with high sensitivity and resolution. Sixteen children, referred for brain imaging in the workup of seizure disorder, were included since they turned out negative after a 1-yr follow-up. A standardized brain presentation was obtained after reslicing and reorienting of the three-dimensional volumetric dataset. RESULTS: Quantitative analysis did not reveal significant left-right uptake differences per patient. Three age clusters were investigated that showed differences in regional uptake, mainly a relatively increased uptake in basal ganglia, visual and motor cortex. An uptake ratio or perfusion index was calculated after normalization. Normal limits were established for the children in the three groups. CONCLUSION: Technetium-99m-ECD is a safe agent for children and should be the radiopharmaceutical of choice for brain perfusion studies because of favorable radiation dosimetry and stability. The age dependence of perfusion necessitates a database comparison before concluding that the observed perfusion pattern is normal. PMID- 9225802 TI - Brain perfusion SPECT in Lyme neuroborreliosis. AB - SPECT imaging brain perfusion using 99mTc-HMPAO was performed on a 38-yr-old women with Lyme neuroborreliosis confirmed by autopsy. The patient had been suspected of spinocerebellar degeneration. Cerebral blood flow was diffusely decreased throughout cerebral cortices but cerebellar blood flow was not impaired, which indicated that the diagnosis was unlikely spinocerebellar degeneration. These findings suggested that brain perfusion SPECT provides useful information in diagnosing the patients with Lyme neuroborreliosis, especially when spinocerebellar degeneration is included in the differential diagnosis. PMID- 9225803 TI - Anterior operculum syndrome localized by SPECT. AB - The aim of this case report was to present a patient with complete anarthria and orofacial apraxia without other relevant neurological deficit. The clinical features are compatible with anterior operculum syndrome. METHODS: A regional brain perfusion scan was done using 99mTc-HMPAO and a SPECT gamma camera. A brain CT scan and an MRI were also performed. RESULTS: Brain CT and MRI were not diagnostic. On brain SPECT, hypoperfusion of the left inferior area of the frontal lobe was noted. CONCLUSION: The patient studied showed an uncommon case of anterior operculum syndrome of focal degenerative origin localized by SPECT. SPECT may be a useful and effective method for diagnosis of this unusual neurological deficit. PMID- 9225804 TI - Technetium-99m-meso-HMPAO as a potential agent to image cerebral glutathione content. AB - To clarify whether the content of glutathione (GSH) in the brain can be estimated by the uptake of 99mTc-meso-HMPAO, we conducted the following in vivo and in vitro experiments. METHODS: We investigated the effect of diethyl maleate (DEM) and buthionine sulfoximine (BSO) administration on the brain uptake of 99mTc-meso HMPAO in the mouse, rat and rabbit, and the chemical specificity of in vitro interaction of 99mTc-HMPAO to GSH using measurements of octanol-extractable radioactivity as an index of remaining intact tracer. RESULTS: The uptake of 99mTc-meso-HMPAO in the mouse and rat brain were reduced together with decreased content of GSH by preloading of DEM, a GSH depletor that acts through glutathione S-transferase. Neither 99mTc-meso-HMPAO uptake nor GSH content was affected in the rabbit brain. Similarly, the uptake of 99mTc-meso-HMPAO and GSH content in the mouse brain was reduced by preinjection of BSO, a GSH depletor that acts through gamma-glutamylcysteine synthetase. In an in vitro study, 99mTc-HMPAO showed reactivity to the molecules possessing a -SH group, but were not specific to GSH. The order of 99mTc-meso-HMPAO reactivity to the mouse brain homogenate agreed with the order of GSH concentration: normal > BSO > DEM. GSH was a major contributor to the conversion reaction of 99mTc-meso-HMPAO to hydrophilic complex in mouse brain homogenate. CONCLUSION: GSH may have a major responsibility for trapping 99mTc-HMPAO in the brain, suggesting the possibility of in vivo measurement of brain GSH with 99mTc-meso-HMPAO. PMID- 9225805 TI - Hyperfixation of copper-62-PTSM in rat brain after transient global ischemia. AB - We evaluated the regional distribution of 62Cu-pyruvaldehyde bis(N4 methylthiosemicarbazone) (62Cu-PTSM), a potential PET perfusion agent, in the rat brain and observed hyperfixation in transient global ischemia in rats. METHODS: The distribution of 62Cu-PTSM was examined in comparison with that of 123I labeled p-iodophenyl-N-isopropylmethanphetamine (123I-IMP) as a reference blood flow marker. Brain uptake of these two tracers was measured in Wistar rats subjected to 30-min four-vessel occlusion followed by recirculation for 10 min, 1 hr or 1, 3 or 5 days. Tracers were injected intravenously into rats 10 min before decapitation. The activities of Complex I and Complex I-III of mitochondria and the concentration of sulfhydryl (SH) groups were also measured. RESULTS: Copper 62-PTSM showed accelerated accumulation in the brain at 1 hr and 1 day after reperfusion when compared with that of 123I-IMP (p < 0.01), and this enhancement was considered to be due to hyperfixation. At these time points, SH concentration was significantly decreased (p < 0.01). On the other hand, the activity of Complex I was not influenced by ischemia/reperfusion, but that of Complex I-III was decreased to 65-70% of the control level (p < 0.01). CONCLUSIONS: Copper-62 PTSM showed hyperfixation most possibly as a result of increased NADH concentration, caused by disturbed electron transport in mitochondria. PMID- 9225806 TI - Scintigraphy with indium-111-labeled homologous (donor) platelets in the platelet transfusion refractory bone marrow transplant patient. AB - Some bone marrow transplant patients who require multiple platelet transfusions as a consequence of post-transplant thrombocytopenia become refractory to these transfusions. As the spleen is the primary site of destruction for senescent and damaged platelets, splenectomy is a potential therapy for persistent thrombocytopenia. Scintigraphy with 111In-labeled platelets has been used to identify increased splenic sequestration and destruction in various platelet disorders, especially idiopathic thrombocytopenic purpura, before consideration of therapeutic splenectomy, but this technique has not been widely described in platelet transfusion refractory bone marrow transplant patients. We report on the results of 111In-labeled platelet scans in two such patients and review the pertinent literature in relation to the possible benefits and limitations of this scanning technique. PMID- 9225807 TI - Indium-111-leukocyte imaging: a case of peritonitis mimicking inflammatory bowel disease. AB - Leukocytes labeled with 99mTc-HMPAO and 111In have been used extensively in imaging inflammatory disorders, including inflammatory bowel disease (IBD), which has the appearance of tubular bowel activity. Peritonitis is inflammation of the serosal surfaces lining the peritoneal cavity which envelopes the bowel, giving a pattern of diffuse abdominal uptake on imaging. We present a case of an elderly man with surgically and pathologically confirmed peritonitis whose preoperative leukocyte scan mimicked the findings of IBD. Our findings suggest that diffuse peritonitis can mimic IBD on an 111In-leukocyte scan. PMID- 9225808 TI - Methodological validation and clinical usefulness of carbon-14-urea breath test for documentation of presence and eradication of Helicobacter pylori infection. AB - A simple [14C]urea breath test (C-14-UBT) was validated with aims of determining accuracy in documenting both the presence and proof of eradication of Helicobacter pylori infection. METHODS: Fifty-six dyspeptic patients had endoscopy with biopsies and C-14-UBT. Eleven biopsy-proven H. pylori-negative patients allowed C-14-UBT normal value determination. Forty-three patients with recurrent peptic ulcer disease and biopsy-proven H. pylori infection were included in an antimicrobial eradication protocol. Endoscopy with biopsies and C 14-UBT were done again 8 wk after initiation of treatment in 35 patients. For C 14-UBT, 185 kBq (5 microCi) of [14C]urea was swallowed. Breath samples obtained up to 20 min were counted to calculate AS20, [(% 14CO2 dose excreted/mmol of CO2) x kg] at 20 min. Combined histologic and microbiologic analyses of antral biopsies were used as a gold standard. RESULTS: The positivity value was set as AS20 > 0.33% (mean + 3 s.d. of AS20 in H. pylori-negative patients). Diagnosis of H. pylori infection was correct with C-14-UBT in 55/56 patients (44 true positive, 11 true-negative and 1 false-negative; sensitivity = 98%; specificity = 100%). As a proof of eradication, C-14-UBT correctly classified 33/35 patients (5 true-positive, 28 true-negative and 2 false-positive; sensitivity = 100%; specificity = 93%). The C-14-UBT global performance yielded sensitivity, specificity and accuracy of 98%, 95% and 97%, respectively. A significant correlation (r = 0.84) was found between AS20 and the number of H. pylori colonies on culture. CONCLUSION: This C-14-UBT is highly accurate both for diagnosis and proof of eradication of H. pylori infection and reflects the antral bacterial load. It is simple, fast and inexpensive, and it is therefore suitable for clinical practice. PMID- 9225809 TI - The exercise renogram and its interpretation. AB - The exercise renogram is a rarely used diagnostic procedure, but it may visualize an exercise-induced change in renal function related to the pathophysiology of essential hypertension, which could greatly increase interest in this examination. The aim of this study was to demonstrate the interpretative approach and the terminology which is used to describe results of exercise renography, using a population of hypertensives with renovascular disease. METHODS: We reviewed the examinations of 70 hypertensives who had supine renography as well as exercise renography with a 60-80 W work load. Forty-eight patients were examined with 99mTc-MAG3 and 22 with 131I hippurate. The renographic and angiography results were recorded as well as the antihypertensive drugs used and the site of vascular lesions. RESULTS: Thirty-three hypertensives developed a bilateral-abnormal exercise renogram, which appears to be associated with primary hypertension. Eight individuals responded to exercise with a unilateral-abnormal exercise renogram, in a kidney behind a stenosis. Only 19 patients had a normal exercise renogram, and 10 had only one functioning kidney. Pathology recognized but unrelated to the intervention included nonfunctioning and small kidneys and pelvic retention. CONCLUSION: Exercise renography's only indication is for recognition of pathology unique to hypertension, since other function disturbances were recognized in resting renograms. PMID- 9225810 TI - Scintigraphic evidence of pulmonary vascular occlusion in sickle cell disease. AB - The acute chest syndrome of sickle cell disease is believed to be primarily a microvascular event. It will rarely involve the larger pulmonary vasculature. We present a case of sickle cell disease where segmental pulmonary arteries were temporarily occluded during the episode of sickling. PMID- 9225811 TI - Splenogonadal fusion diagnosed by spleen scintigraphy. AB - Splenogonadal fusion (SGF) is a rare congenital malformation characterized by fusion of the spleen and a gonad (almost always the left one) frequently associated with orofacial and/or limb developmental abnormalities. Only 125 cases were reported between 1883 and 1994. This report concerns a case of SGF in a 20 yr-old woman with an accidental finding of a splenic space-occupying lesion protruding into the lower abdomen in ultrasound and CT. Radiocolloid spleen scintigraphy and SPECT proved to be the best procedure to establish the correct diagnosis of SGF. As SGF is often asymptomatic, more liberal use of splenic scintigraphy is suggested in patients with congenital limb and/or orofacial anomalies. SGF should be included among the differential diagnoses of left abdominal, pelvic or scrotal masses. PMID- 9225812 TI - Copper-62-ATSM: a new hypoxia imaging agent with high membrane permeability and low redox potential. AB - An ideal hypoxia imaging agent should have high membrane permeability for easy access to intracellular mitochondria and low redox potential to confer stability in normal tissue, but it should be able to be reduced by mitochondria with abnormally high electron concentrations in hypoxic cells. In this context, nitroimidazole residues are not considered to be essential. In this study, Cu(II) diacetyl-bis(N4-methylthiosemicarbazone) (Cu-ATSM), a 62Cu-bisthiosemicarbazone complex, with high membrane permeability and low redox potential, was evaluated as a possible hypoxia imaging agent, using electron spin resonance spectrometry and the Langendorff isolated perfused rat heart model as well as rat heart left anterior descending occlusion model. METHODS: Nonradioactive Cu-ATSM was incubated with rat mitochondria, after which reduction of Cu(II) to Cu(I) was measured with electron spin resonance. As a model of hypoxic mitochondria, rotenone (Complex I inhibitor)-treated mitochondria were used. RESULTS: In this study, Cu-ATSM was reduced by hypoxic but not by normal mitochondria. CONCLUSION: Thus, retention of 62Cu-ATSM was studied serially in perfused rat hearts under conditions of normoxia (95% O2 + 5% CO2), hypoxia (95% N2 + 5% CO2) and reoxygenation (95% O2 + 5% CO2). In normoxia and reoxygenation, 62Cu-ATSM injected as a single bolus showed low retention (23.77% and 22.80%, respectively) 15 min after injection, but retention was increased markedly under hypoxic conditions (81.10%). Also, in the in vivo left anterior descending occluded rat heart model, 62Cu-ATSM retention was inversely correlated with accumulation of 201Tl, a relative myocardial blood flow marker. PMID- 9225813 TI - Physiologic smoothing of blood time-activity curves for PET data analysis. AB - Blood or plasma time-activity curves (TACs) are used as the input function for mathematical models of tracer kinetics in several applications including PET. Uncertainty associated with both the blood data and the PET tissue data can result in uncertainty in the estimates of metabolic rates, blood flow, etc. METHODS: This article presents an approach to reduce the uncertainty in the blood TAC by fitting a model to the curve. The model includes a choice of bolus or infusion input and has three compartments (plasma, interstitial fluid and tissue fluid) with exchange between them. There is a parameter for loss from the plasma compartment. To test the utility of smoothing blood TACs with this approach, a program was set up, using the fluorodeoxyglucose (FDG) model, with simulated noisy blood and tissue TACs. The smoothed blood TAC was compared to a linearly interpolated TAC as the input function with a compartmental model parameter estimation program and with graphical analysis. RESULTS: With a well sampled blood TAC (19 points), the model approach is somewhat more accurate than linear interpolation if the s.d. of noise added to the data exceeded 10%. With sparsely sampled blood TACs (five points) or with a large gap in the blood TAC, the modeled approach was markedly better. For graphical analysis, the model smoothed TAC was also more accurate, although, in general, the results were not as sensitive to the input function. CONCLUSION: This approach, using a physiologically reasonable model to smooth the blood TAC, is a useful aid in PET data analysis, particularly when the data are quite noisy or when there are large gaps in the data. PMID- 9225814 TI - Ultrasound guided internal radiotherapy using yttrium-90 glass microspheres for liver malignancies. PMID- 9225815 TI - Technetium-99m-sestamibi cellular uptake: passive or secondary active transport? PMID- 9225816 TI - BMIPP and flow tracers in myocardial hypoperfusion. PMID- 9225817 TI - Antigranulocyte antibody uptake in bone marrow is age-dependent. PMID- 9225818 TI - Effectively using evidence for decision analysis. PMID- 9225819 TI - Decision tree sensitivity analysis for cost-effectiveness of FDG-PET in the staging and management of non-small-cell lung carcinoma. PMID- 9225820 TI - Breast cancer consensus report. PMID- 9225821 TI - Absence of short-term effects of glucagon-like peptide-1 and of hyperglycemia on plasma leptin levels in man. AB - In rodents, leptin and the incretin glucagon-like peptide-1 (7-36) amide (GLP-1) affect feeding at least in part via interaction with hypothalamic neuropeptide Y (NPY), suggesting that cross talk may exist between GLP-1 and the ob gene product. Besides insulin, acute hyperglycemia has recently been shown to induce ob gene expression. To address the question of whether leptin plasma levels in humans are affected by GLP-1 infusion and/or hyperglycemia, eight healthy volunteers were studied during euglycemia and hyperglycemic clamping with or without GLP-1 administration while insulin levels were kept constant by somatostatin infusion. Under all conditions, leptin plasma levels remained unchanged, demonstrating that in humans leptin plasma concentrations are affected neither by short-term peripheral GLP-1 infusion nor by hyperglycemia, which suggests that postprandial GLP-1 release and hyperglycemia do not modulate secretion of the ob gene product. PMID- 9225822 TI - Corticotropin increases the receptor-specific uptake of native low-density lipoprotein (LDL)--but not of oxidized LDL and native or oxidized lipoprotein(a) [Lp(a)]--in HEPG2 cells: no evidence for Lp(a) catabolism via the LDL-receptor. AB - To understand the interaction of corticotropin (ACTH) and lipid catabolism, we analyzed the influence of ACTH on receptor-mediated lipoprotein uptake and compared the uptake and degradation of human native (N-LDL) and oxidized (Ox-LDL) low-density lipoprotein and native (N-Lp(a)) and oxidized (Ox-Lp(a)) lipoprotein(a) by human hepatoma (HepG2) cells. The receptor affinity of N-LDL, Ox-LDL, N-Lp(a), and Ox-Lp(a) was comparable (Kd, 33, 13, 24, and 13 micrograms/mL medium), whereas the maximum degradative capacity was 10.5-fold higher in N-LDL (Vmax, 1,978 ng/mg cell protein) compared with Ox-LDL (189 ng/mg). In N-LDL, it was 4.5-fold higher than in N-Lp(a) (442 ng/mg) and eightfold higher than in Ox-Lp(a) (246 ng/mg) (P < .05). Addition of ACTH to the cell cultures increased receptor-specific degradation of N-LDL by 44% (2,866 v 1,978 ng/mg, P < .05), whereas changes in Ox-LDL, N-Lp(a), and Ox-Lp(a) showed no significant increase. No differences in uptake specificity were observed with or without ACTH. In addition, a 12-hour preincubation of liver cells with LDL increased Lp(a) uptake by 40% to 50% with (411 v 620 ng/mg) and without (393 v 558 ng/mg) ACTH administration. These data indicate that ACTH elevates receptor specific uptake of N-LDL, but only to a low extent versus Ox-LDL, N-Lp(a), or Ox Lp(a). These results support the hypothesis that catabolism of oxidized lipoproteins and Lp(a) through the LDL receptor pathway is only a minor route of lipid metabolism, whereas LDL receptor activity itself can be stimulated by ACTH. PMID- 9225823 TI - Decreased substance P content in the rectal mucosa of diabetics with diarrhea and constipation. AB - Substance P (SP), vasoactive intestinal polypeptide (VIP), and somatostatin content in rectal mucosa were determined by radioimmunoassay (RIA) in 38 diabetic patients (12 with normal bowel function, 13 with diabetic diarrhea, and 13 with constipation) and in 10 nondiabetic controls with normal bowel function. SP content (picograms per milligram) in the rectal mucosa of diabetics with normal bowel function was significantly higher than that of nondiabetic controls (P < .05). SP content in the rectal mucosa of diabetics with diabetic diarrhea and constipation was significantly lower than in diabetics with normal bowel habits and nondiabetic controls (P < .05). No differences were found in the rectal mucosa content of VIP and somatostatin between the different groups of diabetics and controls. Diabetic diarrhea is a condition with an intermittent nature and frequently alternates with constipation. Our findings showing low levels of rectal mucosa SP in both conditions suggest a possible common role of SP in the pathogenesis of diabetic diarrhea and constipation. PMID- 9225824 TI - Elevated serum lipoprotein(a) levels in young women with endometriosis. AB - Elevated serum lipoprotein(a) [Lp(a)] levels increase the risk of cardiovascular disease if levels of low-density lipoproteins (LDLs) are also high. The biological function of Lp(a) is unknown, but plasma levels may be elevated in inflammatory disease. Endometriosis is a common gynecologic disorder in which endometrial tissue is found outside of the lining of the uterine cavity. There is an immune component to this condition whereby the number of peritoneal macrophages is increased and the level of prostanoids and cytokines in peritoneal fluid is elevated. In the present study, we measured serum lipid, lipoprotein, and apolipoprotein levels in 29 women with endometriosis and in 29 matched healthy controls. Fasting serum triglyceride and apolipoprotein (apo) Al levels were higher in women with endometriosis (+28.1%, P < .001, and +12.3%, P < .01, respectively), but there were no significant differences in LDL or high-density lipoprotein (HDL) cholesterol levels. Serum Lp(a) levels were fivefold higher (P < .01) in the patients (median, 15.0 mg/dL; range, 0.05 to 60.0) than in controls (median, 3.1 mg/dL; range, 0.05 to 57.2). The distribution of apo(a) isoforms was similar in the two groups, but in women with endometriosis the individual apo(a) isoforms tended to be associated with higher serum Lp(a) levels. Endometriosis may represent a relatively common condition in which to investigate the role of Lp(a) in human metabolism. PMID- 9225825 TI - Physiological role of the opioid-cholinergic interaction in growth hormone neuroregulation: effect of sex and food intake. AB - Studies performed in animals and humans have suggested a functional interaction between opioid and cholinergic systems in the control of growth hormone (GH) secretion. Moreover, the sex-dependent modulation of GH secretion in humans is well established. To investigate the role of sex and food intake in the regulation of the reciprocal influences of opioids and acetylcholine in the modulation of GH secretion, we studied the GH response to pyridostigmine (PYR) alone and during a naloxone (NAL) infusion in a group of normal men and women before a meal (at 1:00 PM) and postprandially. In women, the response of GH to PYR alone before the meal was significantly lower than in the men (area under the curve [AUC], mean +/- SEM, 320.18 +/- 87.16 v 1,031.06 +/- 333.21 micrograms/L/90 min, P < .01). Before the meal, NAL completely abolished the response of GH to PYR in men (AUC, 1,031.06 +/- 333.21 v 16.50 +/- 7.50 micrograms/L/90 min, P < .01), whereas infusion of NAL did not significantly modify the GH response to PYR in women. Consumption of the meal significantly decreased PYR-induced GH release in both women (AUC, 21.75 +/- 12.75 v 320.18 +/- 87.16 micrograms/L/90 min, P < .05) and men (AUC, 45.75 +/- 18.75 v 1,031.06 +/- 333.21 micrograms/L/90 min, P < .01). Conversely, food intake did not change the effects of NAL infusion on the GH response to PYR either in women or in men. We conclude that the sex-dependent opioid modulation of PYR-induced GH secretion is observed before a meal but not in the postprandial state. Food intake may be hypothesized to influence the cholinergic regulation of GH secretion and the sex-dependent opioid modulation of central cholinergic tone. PMID- 9225826 TI - Extrapancreatic action of truncated glucagon-like peptide-I in Otsuka Long-Evans Tokushima Fatty rats, an animal model for non-insulin-dependent diabetes mellitus. AB - To clarify the mechanism(s) of the antidiabetic effects of truncated glucagon like peptide-1 (GLP-1) in diabetics, we examined its insulinotropic and extrapancreatic effects in a newly established strain of spontaneously non insulin-dependent diabetic (NIDDM) rats, Otsuka Long-Evans Tokushima Fatty (OLETF) rats, that received a continuous infusion of truncated GLP-1 620 pmol/d/kg (G group, n = 12) or of vehicle (V group, n = 12) for 4 weeks by Alzet pump. Nonfasting plasma glucose levels were significantly lower (P < .05) in the G group than in the V group (7.0 +/- 0.67 v 9.1 +/- 1.7 mmol/L), and fasting plasma immunoreactive insulin (IRI) levels were lower in the former than in the latter (0.63 +/- 0.31 v 0.78 +/- 0.25 nmol/L). At day 15 of infusion, the G group showed an attenuated plasma glucose response to an oral glucose load, but had plasma IRI levels comparable to those in the V group. A long-term infusion of truncated GLP-1 increased the glucose infusion rate (GIR) significantly (P < .05) during a euglycemic-hyperinsulinemic clamp test (59.0 +/- 14.8 mumol/kg/min for group G v 38.9 +/- 12.2 for group V), but hepatic glucose output (HGO) did not differ significantly for either group. Uptake of 2-deoxy-D-glucose (2DG) by peripheral muscles in the G group was as much as 2.4-fold higher than in the V group (5.52 +/- 2.04 v 2.29 +/- 0.97 mumol/100 g muscle weight/min). We conclude from these data that truncated GLP-1, in addition to its well-known incretin effect, is capable of augmenting insulin action in peripheral tissues of diabetics, which can contribute, in part, to improve glucose intolerance in OLETF rats. PMID- 9225827 TI - Ethanol exerts acute protein-sparing effects during postabsorptive but not during anabolic conditions in man. AB - Ethanol abuse is frequently associated with protein malnutrition. To assess the acute effects of ethanol on whole-body protein metabolism, [1-13C]leucine kinetics were measured in eight postabsorptive normal male subjects three times, ie, during administration of two doses of ethanol (dose 1, 0.52 g/kg during 2 hours and 0.3 g/kg during 3 hours; dose 2, 0.69 g/kg during 2 hours and 0.3 g/kg during 3 hours) and during saline (controls). During the last 2 hours of the studies, glucose, insulin, and amino acids were infused to assess the effects of ethanol on protein kinetics under anabolic conditions (euglycemic clamp). The decreases in leucine flux (reflecting whole-body protein breakdown) and nonoxidative leucine disappearance (a parameter of protein synthesis) during saline infusion were abolished in both ethanol protocols (P < .05 or less v saline). The rate of leucine oxidation decreased during the higher dose of ethanol compared with saline (P < .005), indicating an anticatabolic effect. During anabolic conditions (clamp), leucine flux and nonoxidative leucine disappearance were significantly higher in both ethanol studies compared with saline (P < .05). Resting energy expenditure (REE) and oxygen consumption (VO2) during the euglycemic clamp increased to a greater degree during both ethanol studies than during saline (P < .05 or less). Thus, an elevation of blood ethanol concentrations to the levels observed in social drinking results in a net anticatabolic effect (diminished leucine oxidation) when ethanol is administered alone. However, during administration of other nutritional substrates, the anticatabolic effect was not detectable, possibly because ethanol enhanced nutrient-induced thermogenesis. PMID- 9225828 TI - Longitudinal changes of biochemical parameters in muscle during critical illness. AB - The study was undertaken to characterize the time course of biochemical parameters in skeletal muscle during critical illness to gain information for the design of a suitable protocol for interventional studies using metabolic or nutritional manipulation. Critically ill patients in our intensive care unit ([ICU] N = 9) were investigated on two separate sampling occasions with percutaneous muscle biopsies for determination of protein, nucleic acids, free amino acids, energy-rich phosphates, fat, water, and electrolytes. The first biopsy specimen was taken 3 to 11 days after admission and the second biopsy specimen 3 to 7 days later. Protein concentration, expressed as alkali-soluble protein (ASP)/DNA, decreased by 12% (P < .02) between the two biopsies. The total free amino acid content was only 50% of normal, but remained unaltered over time. In particular, the concentration of glutamine remained low, approximately 25% of normal. In contrast, branched-chain amino acid (BCAA) increased by 25% (P < .05) and phenylalanine by 55% (P < .05) between biopsies. The fat content related to fat-free solid (FFS) increased by 130% (P < .001) between the two biopsies. Muscle water did not change during the study period. The extracellular portion was double the normal value when related to FFS. Intracellular water, on the other hand, was outside the 95% confidence interval for normal values in the second biopsy. The concentrations of adenosine triphosphate (ATP), creatine, phosphocreatine, and the phosphorylated fraction of total creatine remained at the same level between the two biopsies. We conclude that in critically ill patients, there is a decrease in protein content over time and increases in BCAA, phenylalanine, and fat content, while the low glutamine level and high extracellular water content remain unaltered. The temporal alterations were well characterized after a 5-day study period. PMID- 9225830 TI - Partial preservation of pancreatic beta-cells by vanadium: evidence for long-term amelioration of diabetes. AB - Streptozotocin (STZ)-diabetic rats treated with vanadium can remain euglycemic for up to 20 weeks following withdrawal from vanadium treatment. In this study, we examined the effects of short-term vanadium treatment in preventing or reversing the STZ-induced diabetic state. Male Wistar rats were untreated (D) or treated (DT) with vanadyl sulfate for 1 week before administering STZ. Treatment was subsequently maintained for 3 days (DT3) or 14 days (DT14) post-STZ, after which vanadium was withdrawn. At 4 to 5 weeks post-STZ and following long-term withdrawal from vanadium, DT14 rats demonstrated levels of food and fluid intake and glucose tolerance that were not significantly different from those of age matched untreated nondiabetic rats, and had significantly reduced glycemic levels in the fed state compared with D and DT3 groups. The proportion of animals that were euglycemic (fed plasma glucose < 9.0 mmol/L) was significant in DT14 (five of 10) relative to D (one of 10) and DT3 (one of 10) (P = .01). All euglycemic animals had an improved pancreatic insulin content that, albeit low (12% of control), was strongly linked to euglycemia in the fed state (r = -.91, P < .0001). Moreover, the highly significant correlation persisted with the analysis of untreated STZ-rats alone (r = -.95, P < .0001). Similarly, improvements in glucose tolerance and insulin secretory function in euglycemic rats were strongly correlated with small changes in residual insulin content. Hence, as vanadium pretreatment did not prevent STZ-induced beta-cytotoxicity, the vanadium-induced amelioration of the diabetic state appears to be secondary to the preservation of a functional portion of pancreatic beta cells that initially survived STZ toxicity. The partial preservation of pancreatic beta cells, albeit small in proportion to the total insulin store, was both critical and sufficient for a long-term reversal of the diabetic state. These results suggest that apparently modest effects in preserving residual pancreatic insulin content can have profound consequences on glucose homeostasis and may bear important implications for interventions that have "limited" protective effects on beta cells. PMID- 9225829 TI - Lipoic acid reduces glycemia and increases muscle GLUT4 content in streptozotocin diabetic rats. AB - Alpha lipoic acid (lipoate [LA]), a cofactor of alpha-ketodehydrogenase, exhibits unique antioxidant properties. Recent studies suggest a direct effect of LA on glucose metabolism in both human and experimental diabetes. This study examines the possibility that LA positively affects glucose homeostasis in streptozotocin (STZ)-induced diabetic rats by altering skeletal muscle glucose utilization. Blood glucose concentration in STZ-diabetic rats following 10 days of intraperitoneal (i.p.) injection of LA 30 mg/kg was reduced compared with that in vehicle-treated diabetic rats (495 +/- 131 v 641 +/- 125 mg/dL in fed state, P = .003, and 189 +/- 48 v 341 +/- 36 mg/dL after 12-hour fast, P = .001). No effect of LA on plasma insulin was observed. Gastrocnemius muscle crude membrane GLUT4 protein was elevated both in control and in diabetic rats treated with LA by 1.5- and 2.8-fold, respectively, without significant changes in GLUT4 mRNA levels. Gastrocnemius lactic acid was increased in diabetic rats (19.9 +/- 5.5 v 10.4 +/- 2.8 mumol/g muscle, P < .05 v nondiabetic rats), and was normal in LA-treated diabetic rats (9.1 +/- 5.0 mumol/g muscle). Insulin-stimulated 2-deoxyglucose (2 DG) uptake into isolated soleus muscle was reduced in diabetic rats compared with the control group (474 +/- 15 v 568 +/- 52 pmol/mg muscle 30 min, respectively, P = .05). LA treatment prevented this reduction, resulting in insulin-stimulated glucose uptake comparable to that of nondiabetic animals. These results suggest that daily LA treatment may reduce blood glucose concentrations in STZ-diabetic rats by enhancing muscle GLUT4 protein content and by increasing muscle glucose utilization. PMID- 9225831 TI - Protective effect of D-alpha-tocopherol on the function of human mesangial cells exposed to high glucose concentrations. AB - Altered functions of mesangial cells (MCs) induced by high glucose levels are thought to play an important role in the pathogenesis of diabetic nephropathy. We investigate whether D-alpha-tocopherol (Toc), an antioxidant, can prevent malfunction of cultured human MCs induced by high-glucose media. Incubating MCs with 33 mmol/L glucose caused increased lipid peroxide (LPO) levels, disturbed cell replication, enhanced cytotoxicity, enhanced activity of the diacylglycerol (DAG)-protein kinase C (PKC) pathway, and overproduction of fibronectin and eicosanoids (6-keto prostaglandin F1 alpha [PGF1 alpha] and thromboxane B2 [TXB2]). The amount of LPO in MCs grown in 5 mmol/L glucose was reduced by the addition of Toc in a dose-dependent manner. Since the maximum effect of Toc on decreasing LPO was achieved at a concentration of 100 mumol/L, this dose was selected for the following experiments. Addition of Toc prevented increased LPO levels and [51Cr]-release from MCs induced by high-glucose media without affecting cell number. Toc decreased the total DAG level and PKC activity in membrane fractions in MCs cultured at both 5 and 33 mmol/L glucose. Furthermore, glucose-induced overproduction of fibronectin and eicosanoids from MCs was completely abolished by Toc. These results strongly suggest that Toc ameliorates glucose-induced malfunctions of MCs in vitro. PMID- 9225832 TI - Nutritional and metabolic effects and significance of mild orotic aciduria during dietary supplementation with arginine or its organic salts after trauma injury in rats. AB - The effects of acute food deprivation and subsequent refeeding with isonitrogenous oral liquid diets supplemented with arginine (ARG), ARG alpha ketoglutarate (AKG), or ARG alpha-ketoisocaproate (AKIC) were examined in a Sprague-Dawley rat trauma model (bilateral femur fracture). Both control and trauma rats were starved for 2 days and then pair-fed for 4 days with one of four liquid isonitrogenous diets: diet 1 was a basal casein-based diet, and diets 2, 3, and 4 were the basal diet in which 10% of the nitrogen was replaced by ARG, AKG, or AKIC nitrogen. Two days of starvation resulted in a 13% loss of body weight and also a 27% decrease in the excretion of orotic acid (OA) in control and trauma rats. Although the ARG content of diets 2, 3, and 4 was the same, ARG- and AKIC-supplemented rats excreted significantly (P < .05) more OA than AKG-fed rats. The low level of OA excretion in AKG-fed rats indicates greater use of ARG for metabolic purposes, including efficient urea cycle operation. The metabolic adaptation and nutritional efficacy, i.e., Increased nitrogen retention, larger weight gain, and altered amino acid (AA) metabolism, of AKIC rats seem to be better than in ARG- or AKG-fed rats. PMID- 9225833 TI - High prevalence of mitochondrial diabetes mellitus in Japanese patients with major risk factors. AB - To identify diabetes mellitus caused by the mitochondrial gene substitution at genomic nucleotide pair 3243 (M3243A-->G) we selected 87 diabetic patients with high risk factors such as maternal inheritance and hearing loss. Total DNA was extracted from peripheral leukocytes, and mitochondrial DNA fragments containing M3243A-->G were amplified by polymerase chain reaction (PCR). The amplified fragments were digested with a restriction endonuclease Apa1 and analyzed by agarose gel electrophoresis. The incidence of the M3243A-->G mutation was 4.6% (four of 87) in diabetic patients with maternal inheritance and/or hearing loss. In a subgroup with both maternal inheritance and hearing loss, the incidence of the mutation was as high as 21.4% (three of 14). Cardiac disorders were also present in all four diabetic patients with the mutation. This study suggests that maternal inheritance and hearing loss are useful clinical findings to identify diabetic patients with the mutation, and that cardiac involvement is a high risk factor for the M3243A-->G mutation. PMID- 9225834 TI - A nitrogen-free hypocaloric diet and recombinant human growth hormone stimulate postoperative protein synthesis: fasted and fed leucine kinetics in the surgical patient. AB - Twelve otherwise healthy patients undergoing elective surgery for resection of rectosigmoid adenocarcinoma were randomly allocated to two groups: one group receiving intravenous dextrose 5% 600 to 800 kcal.d-1 (DX, n = 6) and the other group receiving the same amount of dextrose intravenously plus recombinant human growth hormone (DX + rGH, n = 6). Supplementation with rGH started on the day of surgery and continued postoperatively for 5 days. No nitrogen was provided in the diet. This regimen was started 3 days before surgery and continued for 5 days after surgery. Protein kinetics were studied over a period of 8 hours in all patients. Following an overnight fast, a primed constant infusion of L-[1 13C]leucine was maintained for 4 hours (fasted state) and continued for a further 4 hours (fed state) during which 5% beet dextrose (low 13C content) with or without rGH was administered. The isotope studies were performed on the day before surgery and 6 days after surgery. Other measurements included urinary nitrogen excretion, gaseous exchange, and plasma concentrations of insulin, GH, and insulin-like growth factor-I (IGF-I). Addition of rGH to the dextrose diet had a significant positive effect on protein synthesis (P = .02). Surgery was responsible for a significant increase in postoperative whole-body protein breakdown and synthesis and leucine oxidation (P < .01), although lesser changes were observed in the DX group. An interaction between rGH and surgery was associated with a significant increase in protein synthesis (P = .009), but not with changes in either protein breakdown or leucine oxidation. Carbohydrate provision in the form of beet dextrose during the fed state of the isotopic study did not attenuate the significant decrease in protein synthesis (P = .01) or breakdown (P = .003) either before or after surgery, probably reflecting the absence of nitrogen in the diet. No significant interaction was found between rGH and feeding. These results of leucine kinetics indicate that addition of rGH to a low-dextrose intake in the absence of dietary nitrogen can actually promote protein synthesis. The low levels of leucine oxidation could be explained by the fact that amino acids resulting from protein degradation were directed preferentially toward resynthesis of new proteins rather than to oxidative pathways. There was a significant increase in plasma insulin and GH in the group receiving rGH (P < .05). The postoperative plasma concentration of IGF-I did not change in the latter group compared with the DX group, in which IGF-I concentration decreased significantly (P < .05) as part of the response to combined surgery and dietary restriction. Although both IGF-I and insulin are independently capable of stimulating protein synthesis, elevated levels of either hormone or GH itself may primarily modulate protein synthesis, even with a low intake of carbohydrates. PMID- 9225835 TI - Is the plasma uridine level a marker of the overproduction of uric acid? AB - To determine whether the plasma level of uridine can be used to identify patients with gout, the plasma concentration of uridine was determined in patients with gout and normal subjects. Plasma uridine was significantly higher in patients with gout than in normal subjects. It was also significantly higher in patients with gout of the overexcretion (of uric acid) type than in those with gout of the underexcretion type. Plasma uridine was used to classify gout patients into underexcretion and overexcretion types, with a diagnostic accuracy of 92.5%. Results indicate that the plasma uridine concentration may be a marker of uric acid production and can be used to separate hyperuricemia into the overexcretion and underexcretion types. PMID- 9225836 TI - Time of day and glucose tolerance status affect serum short-chain fatty acid concentrations in humans. AB - Short-chain fatty acids (SCFA) are derived from endogenous (metabolism of fat, carbohydrate, and amino acids) and exogenous (colonic fermentation) sources. To see how time of day and glucose tolerance status influenced serum SCFA concentrations, we determined serum SCFA throughout the day in 22 subjects with impaired glucose tolerance (IGT) and 10 young and eight middle-aged normal controls. On 1 day, insulin sensitivity was assessed as the steady-state plasma glucose (SSPG) level achieved during intravenous infusion of glucose insulin, and somatostatin. On another day, plasma glucose and insulin and serum SCFA levels were measured 12 times over 12 hours with subjects eating a standard diet. SSPG in young controls (5.5 +/- 1.1 mmol/L) was less than in middle-aged controls (9.3 +/- 1.6 mmol/L), which in turn was less than in IGT subjects (13.7 +/- 0.6 mmol/L; P < .01). Mean plasma glucose in IGT subjects was greater than in normal controls, and mean plasma insulin in IGT subjects was higher than in young controls but similar to the levels in middle-aged controls. Mean 12-hour serum acetate in young controls (143 +/- 13 mumol/L) was greater than in middle-aged controls (104 +/- 11 mumol/L) and IGT subjects (113 +/- 5 mumol/L; P < .05). Mean 12-hour serum propionate in young controls (3.8 +/- 0.5 mumol/L) was less than in IGT subjects (5.4 +/- 0.3 mumol/L; P < .01), with middle-aged controls being intermediate (4.6 +/- 0.3 mumol/L). Both young (1.6 +/- 0.3 mumol/L) and middle aged (1.0 +/- 0.2) controls had lower mean butyrate than IGT subjects (3.1 +/- 0.4 mumol/L; P < .05). Levels of all three SCFA varied significantly during the day, tending to decrease after breakfast and increase transiently after lunch and dinner. It is concluded that both time of day and glucose tolerance status affect serum SCFA levels in nondiabetic humans. The results suggest that serum acetate is derived primarily from colonic fermentation, serum butyrate primarily from endogenous fatty acid metabolism, and serum propionate from both exogenous and endogenous sources. PMID- 9225837 TI - Serum leptin is increased in growth hormone-deficient adults: relationship to body composition and effects of placebo-controlled growth hormone therapy for 1 year. AB - The gene product from the ob gene, leptin, has recently been characterized in humans. The circulating level of leptin is related to body mass index (BMI) and more closely to estimates of total body fat, whereas visceral fat has been reported to be of minor importance. However, it is unknown if leptin is directly regulated by hormones that influence substrate metabolism and body composition. We studied leptin in adult growth hormone (GH)-deficient (GHD) patients substituted with GH treatment for 12 months in a parallel double-blind, placebo controlled study. Twenty-seven GHD adults aged 44.9 +/- 1.9 years underwent anthropometric measurements for determination of regional and total body fat (BMI, waist to hip ratio [WHR], computed tomographic [CT] scan, dual-energy x-ray absorptiometry [DEXA] scan, and bioimpedance analysis [BIA]) before and after 12 months of placebo-controlled GH substitution (2 IU/m2) in a parallel design. The same measurements were performed in 42 healthy adults aged 39.1 +/- 1.7 years. The logarithm of serum leptin levels correlated positively with abdominal subcutaneous fat and total body fat (BIA and DEXA) in untreated GHD patients and healthy subjects. Fasting insulin did not correlate with leptin levels in either of the groups. After 12 months of GH administration, the body composition of GHD patients was significantly changed with respect to a marked decrease in body fat. The relations of leptin to the estimates of body fat were maintained, and leptin was furthermore related to BMI and fasting insulin. In multiple linear regression analyses, additional estimates of visceral adiposity (intraabdominal fat and maximal anterior-posterior diameter determined by CT scan) were significant determinants of leptin in the healthy subjects. The increase in fasting insulin levels during GH substitution correlated negatively with the reduction in leptin levels (r = -.823, P = .003). At baseline, leptin levels were increased in the patients compared with controls in both sexes (women, 21.8 +/- 3.3 v 11.3 +/- 1.4 ng/mL, P = .002; men, 8.1 +/- 1.2 v 4.7 +/- 0.7 ng/mL, P = .008). Leptin levels were similar in GHD patients treated for 12 months compared with healthy controls for both women and men (women, 15.9 +/- 2.3 and 11.3 +/- 1.4 ng/mL, P = .163; men, 7.1 +/- 2.8 and 4.7 +/- 0.7 ng/mL, P = .759). In healthy adults and in GHD patients, leptin levels were significantly higher in women than in men (11.3 +/- 1.4 v 4.7 +/- 0.7 ng/mL, P < .001; 21.8 +/- 3.3 v 8.1 +/- 1.2 ng/mL, P < .001). Gender remained a significant determinant of leptin levels in several models of multiple linear regression analysis also including age, estradiol levels, insulin, and estimates of body fat. We conclude that leptin is increased but not differently regulated in GHD patients compared with normal subjects, and that leptin levels are closely related to estimates of body fat. This relationship is maintained during a decrease in body fat due to GH substitution. PMID- 9225838 TI - Atrial natriuretic peptides increase calcitonin gene-related peptide within human circulation. AB - Long-acting natriuretic peptide (LANP), vessel dilator, and atrial natriuretic factor (ANF) (each infused at 100 ng/kg body weight [BW].min for 60 minutes) increased the circulating concentration of calcitonin gene-related peptide (CGRP) threefold to fourfold in 30 healthy humans. Within 30 minutes of stopping ANF infusion, the CGRP circulating concentration had returned to preinfusion levels, whereas its increase secondary to the other atrial peptides was still significant 2 to 3 hours after cessation of their infusions. There was a 50% decreased excretion (P < .001) of CGRP into the urine secondary to LANP and vessel dilator, which correlated with the increase of CGRP in the circulation. The ANF-induced 50% decreased CGRP excretion occurred after the circulating concentration of CGRP had returned to preinfusion levels. Kaliuretic peptide did not affect CGRP circulating concentration or excretion into urine. These data suggest that LANP and vessel dilator inhibit the metabolic breakdown of CGRP as part of their mechanism of increasing CGRP in plasma, whereas the ANF effect of increasing CGRP in plasma appears to be secondary to stimulating the release of CGRP. PMID- 9225839 TI - Effects of aging on dehydroepiandrosterone sulfate in relation to fasting insulin levels and body composition assessed by bioimpedance analysis. AB - Insulin can inhibit dehydroepiandrosterone (DHEA) biosynthesis in humans, as suggested by several studies performed in induced or spontaneous hyperinsulinemia. The increased insulin resistance documented throughout aging, with its accompanying hyperinsulinemia, may contribute to the age-related decline in DHEA synthesis. The aim of this study was to assess if the aging-related differences in DHEA sulfate (DHEA-S) serum levels can be associated with differences in fasting insulin levels, as well as body composition. Two hundred fifty-two healthy subjects of both sexes aged 19 to 90 years with a body mass index (BMI) less than 30 (mean +/- SD, 23.5 +/- 2.4) were studied DHEA-S and insulin serum levels were determined by a radioimmunologic procedure; body composition was assessed by anthropometry (fat mass percentage [FM%] estimated from four skinfold thicknesses by Durnin and Womersley and Siri equations [FM% SKF]) and by bioimpedance analysis (BIA) (FM% estimated by equations developed by Segal et al and Deurenberg et al for subjects < and > 62 years, respectively [FM% BIA]). DHEA-S levels were significantly and inversely related to age in both sexes. No significant aging-related differences were found in fasting insulin levels, although a trend toward an increase was apparent in the women on simple regression analysis. No significant associations were found between DHEA-S and insulin levels. As for body composition, a positive relationship to age was apparent for FM%-SKF, FM%-BIA, and waist to hip ratio (WHR), whereas BMI and phase angle ([PA] a bioelectric parameter considered an index of the ratio between intracellular and extracellular water) were inversely related to age. Fasting insulin levels were positively related to FM% as estimated by both BIA and anthropometry, independently of age in both sexes; in addition, a positive correlation with WHR and with the subscapular to triceps skinfold thickness ratio (SS/TS) was found in men and women, respectively. No significant correlation was apparent between DHEA-S and body composition indices in men, whereas in women a slight negative correlation between DHEA-S and WHR was documented, and was still significant after adjustment for age and fasting insulin. Stepwise multiple regression analysis confirmed that DHEA-S levels are not related to fasting insulin, but are independently related to age and, in women only, to WHR. Our study suggests that the DHEA-S decline due to aging is independent of fasting insulin, at least in healthy, non-obese people. In addition, it is not related to the aging-dependent changes in body composition in terms of FM% and fat-free mass (FFM) percentage (FFM%). Only in women could changes in fat distribution be slightly associated with DHEA-S decline, although such a relation cannot be accounted for by changes in insulin levels. PMID- 9225840 TI - Resistance of lipoproteins from continuous ambulatory peritoneal dialysis patients to in vitro oxidation. AB - Patients with end-stage renal failure on continuous ambulatory peritoneal dialysis (CAPD) develop abnormalities in plasma lipoproteins that may contribute to their increased risk for atherosclerosis. The oxidative modification of lipoproteins is considered to play a central role in atherogenesis. This study examines the susceptibility to oxidation in vitro of low- and high-density lipoprotein (LDL and HDL, respectively) obtained from long-term CAPD patients. CAPD LDL was less susceptible to copper-mediated protein derivatization (fluorescence) compared with control LDL CAPD LDL and HDL displayed less copper promoted conjugated-diene production and lipid peroxide generation, suggesting a greater resistance of CAPD lipoprotein lipids to oxidation. Autooxidation during long-term storage was also much lower in CAPD LDL and HDL. However, when 2,2' azobis(2-amidinopropane) dihydrochloride (ABAP) was used to initiate oxidation, there was no difference in conjugated-diene generation between CAPD and the control. CAPD LDL contained slightly less oxidizable, polyunsaturated fatty acid, but the vitamin E content of CAPD and control LDL was equivalent. Our findings indicate that lipoproteins from uremic patients undergoing long-term CAPD are more resistant to in vitro oxidation than control lipoproteins. PMID- 9225842 TI - Rapid identification of Smith-Lemli-Opitz syndrome homozygotes and heterozygotes (carriers) by measurement of deficient 7-dehydrocholesterol-delta 7-reductase activity in fibroblasts. AB - To extend the enzyme deficiency in Smith-Lemli-Opitz syndrome (SLOS) to extrahepatic tissues, 7-dehydrocholesterol-delta 7-reductase activity was measured in fibroblasts from 10 controls, five SLOS homozygotes, and five obligate heterozygotes. In cells grown almost to confluence in cholesterol containing medium (4 mg/dL), the conversion of [1,2-3H]7-dehydrocholesterol to cholesterol (7-dehydrocholesterol-delta 7-reductase activity) was 3.8 times higher in control than in homozygote cells and 2.2 times higher than in heterozygote cells. After 24 hours' exposure of the fibroblasts to cholesterol deficient medium supplemented with lovastatin, 7-dehydrocholesterol-delta 7 reductase activity increased twofold in controls, but did not change significantly in either heterozygous or homozygous cells. In contrast, the activities of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase and lathosterol 5-dehydrogenase, two key enzymes that precede 7-dehydrocholesterol delta 7-reductase in the cholesterol biosynthetic pathway, and low-density lipoprotein (LDL) receptor-mediated binding were equal in control, homozygote, and heterozygote fibroblasts. Further, HMG-CoA reductase activity and LDL receptor-mediated binding increased after exposure of the cells to cholesterol deficient medium. Fibroblast cholesterol concentrations were approximately equal, although homozygote cells contained 30 times more 7-dehydrocholesterol. Thus, markedly reduced 7-dehydrocholesterol-delta 7-reductase activity that cannot be upregulated after exposure of the cells to cholesterol-deficient medium is diagnostic for the biochemical defect in SLOS. Significantly reduced enzyme activity between the levels in controls and homozygotes without accumulation of 7 dehydrocholesterol in fibroblasts identified heterozygotes. PMID- 9225843 TI - Attitudes toward standardized data collection. AB - BACKGROUND AND PURPOSE: Clinical databases will gain importance as physical therapists need to determine the effectiveness and justify the expense of treatment. Obstacles impeding participation in standardized data collection need to be identified and addressed to construct high-quality databases. SUBJECTS: Subjects were clinicians involved in patient care in a multisite physical therapy/occupational therapy practice that had initiated a standardized data collection program. METHODS: Questionnaires, asking about participation in the data collection project, were distributed to participating clinics. Factor analysis identified attitudes toward standardized data collection. Relationships between efforts to learn standardized methods, attitudes, clinic and clinician characteristics, and levels of participation in the project were investigated. RESULTS: Most respondents (66%) had made some effort to learn the standardized procedures. Five attitudes were identified: inconvenience, acceptance of operational definitions, automation, paperwork, and training. CONCLUSION AND DISCUSSION: Inconvenience of data collection and training in standardized methods need to be addressed to construct large, high-quality databases. PMID- 9225841 TI - Impaired nonoxidative glucose metabolism in patients with liver cirrhosis: effects of two insulin doses. AB - Glucose intolerance is encountered in the majority of cirrhotic patients. This alteration has been attributed to a defective insulin-mediated glucose uptake in peripheral tissue, where nonoxidative glucose disposal seems to be chiefly impaired. To further investigate insulin action under euglycemic conditions, we studied how physiological (100 microU/mL) and pharmacological (1,000 microU/mL) plasma insulin concentrations affect whole-body insulin-mediated glucose uptake, as well as oxidative and nonoxidative glucose disposal, in cirrhotic patients and controls. To this aim, a sequential two-step insulin euglycemic clamp combined with indirect calorimetry was performed in eight cirrhotic patients and six control subjects. During the first step of the clamp, total glucose uptake was reduced by 40% in cirrhotic patients versus controls (4.42 +/- 1.39 v 7.63 +/- 1.60 mg/kg/min, P = .002). By increasing insulin to pharmacological levels, glucose disposal increased in both groups. However, the maximum rate of glucose metabolism achieved in cirrhotic patients was lower than in controls at all times (10.29 +/- 2.04 v 12.82 +/- 0.51 mg/kg/min, P = .012). Glucose oxidation was lower in cirrhotics in the basal state, but similar in both groups during insulin/glucose infusion. On the other hand, the reduced nonoxidative glucose disposal observed in cirrhotic patients was not normalized even by increasing insulin to pharmacological levels. In conclusion, in liver cirrhosis a reduced insulin sensitivity is associated with a reduced insulin responsiveness that is mainly caused by defective nonoxidative glucose disposal. PMID- 9225844 TI - The sensitivity and specificity of pain response to activity and position in categorizing patients with low back pain. AB - BACKGROUND AND PURPOSE: The purposes of this study were to develop screening tests for four low back pain (LBP) diagnoses based on patient reports concerning the severity of pain in various positions and during activities and then to examine the accuracy of these tests in assigning subjects to one of four LBP diagnostic categories. The accuracy of screening tests is determined by calculating sensitivity and specificity and is well established in epidemiology. SUBJECTS: One hundred six consecutive patients who either were being treated for LBP for the first time or had not received medical care for LBP at the participating clinics within the 12 months prior to the study were recruited. METHODS: Subjects completed a Pain Response to Activity and Position Questionnaire at the time of their initial clinic visit. The diagnosis of LBP was obtained from the medical record after at least 1 month of follow-up and the completion of diagnostic testing. Data analysis yielded symptom clusters that were used to produce screening tests for each of the four categories of LBP. RESULTS: Sensitivity, specificity, and positive and negative predictive power of the screening tests were (1) benign back disease: .57,.71,.40, and .82, (2) disk disease: .65,.49,.35, and .77, (3) spinal stenosis: .52, .74, .36, and .85, and (4) disk disease with spinal stenosis: .81, .54, .24, and .94. CONCLUSION AND DISCUSSION: Although more than half of the subjects with a particular LBP diagnosis tested positive for that diagnosis, approximately two thirds of the subjects who tested positive for each of the diagnoses actually had another diagnosis. Negative tests may be more useful in that between 77% and 94% of the subjects without the diagnosis tested negative. Although patient reports of LBP response to position and activity are not sufficient for diagnosis, they may be useful in ruling out a particular diagnosis. PMID- 9225845 TI - Patient sexual behaviors and sexual harassment: a national survey of physical therapists. AB - BACKGROUND AND PURPOSE: The objective of this study was to describe the extent to which physical therapists have experienced patient sexual behaviors and sexual harassment. SUBJECTS AND METHODS: A questionnaire was mailed to 750 individuals selected from a computer-generated, randomized sample provided by the American Physical Therapy Association of members who identified themselves as being licensed physical therapists and currently engaged in clinical practice. Completed questionnaires were returned by 48.6% of the eligible sample. RESULTS: Eight-six percent of the respondents reported having experienced some form of patient sexual behavior in the course of practice. The vast majority of these incidents were not rated as harassment. Sixty-three percent of the respondents, however, reported at least one incident of sexual harassment. Despite those high rates, less than one third of the respondents had received any training in how to handle sexual harassment. CONCLUSION AND DISCUSSION: The results support the findings of previous investigators who concluded that patient sexual behavior and sexual harassment is an important issue that needs to addressed in training programs. PMID- 9225846 TI - Use of the standard error as a reliability index of interest: an applied example using elbow flexor strength data. AB - The intraclass correlation coefficient (ICC) and the standard error of measurement (SEM) are two reliability coefficients that are reported frequently. Both measures are related; however, they define distinctly different properties. The magnitude of the ICC defines a measure's ability to discriminate among subjects, and the SEM quantifies error in the same units as the original measurement. Most of the statistical methodology addressing reliability presented in the physical therapy literature (eg, point and interval estimations, sample size calculations) focuses on the ICC. Using actual elbow flexor make and break strength measurements, this article illustrates a method for estimating a confidence interval for the SEM, shows how an a priori specification of confidence interval width can be used to estimate sample size, and provides several approaches for comparing error variances (and square root of the error variance, or the SEM). PMID- 9225848 TI - Physical therapy implications following the TRAM procedure. AB - The TRAM procedure has gained popularity over the last decade as an autogenous technique for breast reconstruction. Several outcome studies have demonstrated complications from this procedure, as described in this article. Women should be informed about the possible complications prior to surgery. Physical therapists can play an important role in rehabilitation and education for patients who are planning to undergo or who have undergone the TRAM procedure. It is important that physical therapists become well acquainted with the surgical procedure and treatment guidelines to effectively treat patients who have undergone the TRAM procedure, especially as this procedure increases in popularity. In addition, it is important that further research be conducted to substantiate the valuable clinical contribution that physical therapy has on successful recovery following the TRAM operation. PMID- 9225847 TI - Multidimensional assessment of motor function in a child with cerebral palsy following intrathecal administration of baclofen. AB - This case report describes an 11-year-old boy with spastic diplegia whose reflex status, range of motion (ROM), strength, and motor performance were measured before and after implantation of an indwelling system for delivery of intrathecally administered baclofen. Before baclofen use, the subject experienced clonus that interfered with walking, needed assistance with transfers, and was unable to independently put on underwear and socks. Measures of spasticity, kinematics and electromyographic activity during voluntary movements, ROM, Gross Motor Function Measure (GMFM) scores, and self-reports of change were obtained at baseline, before and after bolus baclofen injection, during a double-blind placebo-controlled clinical trial of baclofen administration via an indwelling pump, and after 1 and 2 years of baclofen therapy. Spasticity, Babinski reflexes, clonus, strength, and coactivation of antagonist muscles during voluntary movement were decreased shortly after baclofen administration began. Hip and ankle ROM increased, upper-extremity movement speed increased, independence in dressing and transfers improved, and orthoses were discarded. After 1 and 2 years, GMFM scores were 7.8% and 6.4% above baseline, respectively; the subject won a fitness award. After 2 years, ROM was worse than at baseline and concerns regarding hip subluxation arose. Single-joint movement control and independence improved and spasticity decreased during baclofen administration. PMID- 9225849 TI - Characterization and expression of two members of the S-adenosylmethionine decarboxylase gene family in carnation flower. AB - S-adenosylmethionine decarboxylase (SAMDC; EC 4.1.4.50) is one of the key enzymes in polyamine biosynthesis, and the product of its catalytic reaction, decarboxylated S-adenosylmethionine (dcSAM), serves as an aminopropyl donor in the biosynthesis of spermidine and spermine. In order to provide information on the structure and regulation of SAMDC, we have isolated and sequenced two different SAMDC cDNA clones from carnation petals. The nucleotide sequences of CSDC9 and CSDC16 show 78.3% identity, and the deduced amino acid sequences show 81.7% identity and 86.5% similarity [12]. There are several regions with highly conserved sequences among SAMDC cDNAs of potato, spinach, periwinkle, man and yeast. These conserved regions include a cleavage site for the processing of SAMDC proenzyme and a putative PEST sequence that may be relevant to the rapid degradation of SAMDC protein. Carnation SAMDC cDNAs have long transcript leaders of 472 bp and 502 bp for CSDC9 and CSDC16, respectively. Both sequences contain short upstream open reading frames (uORFs) in their 5'-untranslated regions. The CSDC9 uORF is 54 amino acids from 152 to 317 while the corresponding sequence in CSDC16 is 52 amino acids located from 156 to 314 in each 5'-untranslated region. The nucleotide sequences of uORFs in CSDC9 and CSDC16 were 89.9% identical. In vitro transcription/translation experiments showed: (1) each proenzyme of both cDNAs of SAMDC was converted to two polypeptides consisting of a large subunit (calculated as 31,544 Da and 32,537 Da, respectively) and a small subunit (calculated as 9704 and 9041 Da, respectively) after 20 min of translation; (2) the processing occurs rapidly during the translation of protein. But once the translation process is stopped accumulation of the subunits slows and never reaches completion even after 300 min. The processing of carnation SAMDC enzyme is not stimulated by putrescine in in vitro transcription/translation reaction. PMID- 9225850 TI - The maize mitochondrial plasmid RNA b is associated with protein during synthesis but is not encapsidated. AB - RNA b is the most abundant member of a family of autonomously replicating single- and double-stranded RNA plasmids found in maize mitochondria. The extent to which this molecule is associated with proteins was investigated by rate zonal and CsCl equilibrium density gradient centrifugation of clarified lysates of S cytoplasm maize mitochondria. A soluble complex of RNA b, responsible for synthesis of the more abundant (+) RNA b strand in mitochondrial lysates, was identified. The complex had a buoyant density of 1.49 g/cm3, indicating a substantial non-nucleic acids content. The sedimentation coefficient of the complex, however, was only slightly larger than that of deproteinized RNA b. Synthesis of RNA b as well as the larger RNA plasmid, RNA a, was resistant to heparin, suggesting that, for both RNAs, preformed complexes between an RNA template and an RNA-dependent RNA polymerase capable of elongating in vivo preinitiated RNA plasmid strands, were present in the lysate. Only a small fraction of RNA b molecules were bound in the complex; the bulk of RNA b sedimented at the same rate as the deproteinized RNA. Thus, after replication, maize mitochondrial plasmids are not associated with nucleoprotein capsids although their synthesis takes place through ribonucleoprotein replication complexes. PMID- 9225851 TI - Tissue-specific activation of the osmotin gene by ABA, C2H4 and NaCl involves the same promoter region. AB - The gene encoding the antifungal protein osmotin is induced by several hormonal and environmental signals. In this study, tissue-specific and inducer-mediated expression of the reporter gene beta-glucuronidase (uidA) fused to different fragment lengths of the osmotin promoter was evaluated in transgenic tobacco (Nicotiana tabacum). The region of the promoter between -248 to -108 (Fragment A) was found to be essential and sufficient for inducer (abscisic acid (ABA), C2H4 and NaCl)-mediated expression of the reporter gene. Expression of the reporter gene was developmentally regulated and increased with maturity of leaves, stem and flowers. Expression also was tissue-specific being most highly expressed in epidermis and vascular parenchyma of the stem. The regulators ABA, C2H4 and NaCl exhibited tissue-specific induction of this promoter. The promoter was specifically responsive to C2H4 in flowers at virtually all stages of development, but not responsive in these tissues to ABA or NaCl. Conversely, ABA and NaCl were able to induce reporter gene activity using promoter Fragment A in specific tissues of root where C2H4 was unable to induce activity. Further dissection of the promoter Fragment A into fragments containing either the conserved GCC element (PR); PR/AT; or G/AT sequences, and subsequent testing of these fragments fused to GUS in transgenic plants was performed. These experiments revealed that the promoter fragment containing PR element alone, although required, was barely able to allow responsiveness to C2H4. However, significant C2H4-induced activity was obtained with a promoter fragment containing the AT and PR elements together. PMID- 9225852 TI - DNA-binding properties, genomic organization and expression pattern of TGA6, a new member of the TGA family of bZIP transcription factors in Arabidopsis thaliana. AB - The TGA genes encode a family of basic domain-leucine zipper (bZIP) transcription factors that are conserved in higher plants. We have continued to unravel the complexity of this gene family by using a polymerase chain reaction (PCR)-based approach. Taking advantage of the conserved amino acid sequence in the bZIP domain found in all members of this gene family, two degenerate oligonucleotides were synthesized based on the sequence of this region in order to amplify by PCR the analogous genomic fragments from the various TGA loci in Arabidopsis. This approach has led us to the finding of a new member of the TGA gene family, and subsequently the isolation of a gene designated as TGA6. Further characterization of the TGA6 locus confirmed our prediction that the gene structure of this family is remarkably conserved. Genomic Southern blot analysis revealed that TGA6 is a single-copy gene in Arabidopsis. Based on the genomic sequence information, gene specific primers were synthesized for isolating the cDNA that corresponds to the coding region. Subsequently, the cDNA for TGA6 was cloned and sequenced. Gel mobility shift assays with in vitro translated TGA6 protein showed that TGA6 is more like TGA5 in terms of its in vitro DNA-binding properties. The expression of TGA6 in different tissues was estimated by using reverse transcriptase (RT)-PCR and further analyzed in transgenic Arabidopsis lines expressing a TGA6 promoter GUS fusion. TGA6 promoter activity is found primarily in roots of young seedlings. As seedlings develop, TGA6 is expressed in aging cotyledons, mesophyll cells of hydathodes on leaf margins, vascular tissue and trichomes of senescing rosette leaves, but is very low in primary roots of mature plants. High levels of expression persist in young lateral roots and in regions of the primary root where lateral roots are emerging. In flowers, the activity is localized predominantly to mature pollen grains. The expression pattern of TGA6 reported here is strikingly similar to that of an Arabidopsis acidic chitinase gene. Possible biological significance of TGA6 in cellular defense against pathogens and abiotic stress is discussed. PMID- 9225853 TI - An ozone-responsive region of the grapevine resveratrol synthase promoter differs from the basal pathogen-responsive sequence. AB - Stilbene synthase (STS) is an enzyme involved in the biosynthesis of stilbenes, which are synthesized in various plants in response to pathogen attack, UV irradiation or exposure to ozone. We describe analysis of an ozone inducible STS transcript and its corresponding promoter (Vst1), combined with the beta glucuronidase (GUS) reporter gene. A single ozone pulse (0.1 microliter/l, 10 h) resulted in 11-fold GUS expression. Histochemical localization of GUS activity revealed small spots distributed over the whole leaf. Cross-sections of leaf tissue showed that the Vst1 promoter was induced in palisade and spongy parenchyma cells and to a lesser extent in epidermal cells. Deletions at the 5' end showed that a partial promoter sequence between position -430 and -280 constituted the ozone-responsive region, whereas for effective pathogen inducibility sequences from -280 to -140 have been shown to be necessary. PMID- 9225854 TI - The phosphoenolpyruvate carboxylase (ppc) gene family of Flaveria trinervia (C4) and F. pringlei (C3): molecular characterization and expression analysis of the ppcB and ppcC genes. AB - Phosphoenolpyruvate carboxylase (PEPC) is a central enzyme of C4 and CAM photosynthesis but plants, in addition, contain various non-photosynthetic isoforms with characteristic and variable functions. The partial sequence and a detailed expression analysis of the PpcB and PpcC genes which encode non photosynthetic PEPC isoforms in the C4 plant Flaveria trinervia and the C3 species F. pringlei is presented. Southern analyses showed that PpcB and PpcC sequences are most probably of single-copy in the genomes of F. trinervia and F. pringlei. With gene-specific probes the various ppc transcripts could be distinguished unequivocally from one another and the expression patterns of all ppc genes were compared. PpcB and PpcC transcripts of both F. trinervia and F. pringlei were detected preferentially in roots and stems and at low levels in leaves. Their accumulation patterns are thus similar to each other, but different from that of the PpcA genes which in F. trinervia encode the C4 isoform of PEPC. Transgenic analysis of the 5'-flanking regions of the PpcB genes of both F. trinervia and F. pringlei in tobacco revealed that the PpcB promoter/beta glucuronidase reporter genes were preferentially expressed in the phloem and the roots. Comparison of the PpcB promoter/reporter gene with the accumulation pattern of the PpcB transcripts in F. trinervia and F. pringlei suggests that the expression of the PpcB genes is predominantly controlled by transcription. PMID- 9225855 TI - Expression of a wheat ADP-glucose pyrophosphorylase gene during development of normal and water-stress-affected anthers. AB - In wheat (Triticum aestivum L.), water deficit during meiosis in the microspore mother cells (MMCs) induces pollen abortion, resulting in the failure of fertilization and a reduction in grain set. In stressed plants, meiosis in MMCs proceeds normally but subsequent pollen development is arrested. Unlike normal pollen grains, which accumulate starch during the late maturation phase, stress affected anthers contain pollen grains with little or no starch. Stress also alters the normal distribution of starch in the anther wall and connective tissue. To determine how starch biosynthesis is regulated within the developing anthers of stressed plants, we studied the expression of ADP-glucose pyrophosphorylase (AGP), which catalyzes the rate limiting step of starch biosynthesis. Two partial-length cDNAs corresponding to the large subunit of AGP were amplified by RT-PCR from anther RNA, and used as probes to monitor AGP expression in developing anthers of normal and water-stressed plants. These clones, WAL1 and WAL2, had identical deduced amino acid sequences and shared 96% sequence identity at the nucleic acid level. In normal anthers, AGP expression was biphasic, indicating that AGP expression is required for starch biosynthesis both during meiosis and later during pollen maturation. AGP expression in stressed anthers was not affected during the first phase of starch accumulation, but was strongly inhibited during the second phase. We conclude from these results that the reduced starch deposition later in the development of stressed pollen could be the result of a lower expression of AGP. However, this inhibition of AGP expression is unlikely to be the primary cause of male sterility because anatomical symptoms of pollen abortion are observed prior to the time when AGP expression is inhibited. PMID- 9225857 TI - Footprinting of the spinach nitrite reductase gene promoter reveals the preservation of nitrate regulatory elements between fungi and higher plants. AB - Nitrite reductase (NiR) is the second enzyme in the nitrate assimilatory pathway reducing nitrite to ammonium. The expression of the NiR gene is induced upon the addition of nitrate. In an earlier study, a 130 bp upstream region of the spinach NiR gene promoter, located between -330 to - 200, was shown to be necessary for nitrate induction of beta-glucuronidase (GUS) expression in tissue-specific manner in transgenic tobacco plant [28]. To further delineate the cis-acting elements involved in nitrate regulation of NiR gene expression, transgenic tobacco plants were generated with 5' deletions in the -330 to -200 region of the spinach NiR gene promoter fused to the GUS gene. Plants with the NiR promoter deleted to -230 showed a considerable increase in GUS activity in the presence of nitrate, indicating that the 30 bp region between -230 to -200 is crucial for nitrate-regulated expression of NiR. In vivo DMS footprinting of the -300 to -130 region of the NiR promoter in leaf tissues from two independent transgenic lines revealed several nitrate-inducible footprints. Footprinting within the -230 to 181 region revealed factor binding to two adjacent GATA elements separated by 24 bp. This arrangement of GATA elements is analogous to cis-regulatory sequences found in the promoters of nitrate-inducible genes of Neurospora crassa, regulated by the NIT2 Zn-finger protein. The -240 to -110 fragment of the NiR promoter, which contains two NIT2 consensus core elements, bound in vitro to a fusion protein comprising the zinc finger domain of the N. crassa NIT2 protein. The data presented here show that nitrate-inducible expression of the NiR gene is mediated by nitrate-specific binding of trans-acting factors to sequences preserved between fungi and higher plants. PMID- 9225856 TI - Site-directed mutagenesis of the basic residues 321K to 321G in the CP 47 protein of photosystem II alters the chloride requirement for growth and oxygen-evolving activity in Synechocystis 6803. AB - CP 47, a component of photosystem II (PSII) in higher plants, algae and cyanobacteria, is encoded by the psbB gene. Site-specific mutagenesis has been used to alter a portion of the psbB gene encoding the large extrinsic loop E of CP 47 in the cyanobacterium Synechocystis 6803. Alteration of a lysine residue occurring at position 321 to glycine produced a strain with altered PSII activity. This strain grew at wild-type rates in complete BG-11 media (480 microM chloride). However, oxygen evolution rates for this mutant in complete media were only 60% of the observed wild-type rates. Quantum yield measurements at low light intensities indicated that the mutant had 66% of the fully functional PSII centers contained in the control strain. The mutant proved to be extremely sensitive to photoinactivation at high light intensities, exhibiting a 3-fold increase in the rate of photoinactivation. When this mutant was grown in media depleted of chloride (30 microM chloride), it lost the ability to grow photoautotrophically while the control strain exhibited a normal rate of growth. The effect of chloride depletion on the growth rate of the mutant was reversed by the addition of 480 microM bromide to the chloride-depleted BG-11 media. In the presence of glucose, the mutant and control strains grew at comparable rates in either chloride-containing or chloride-depleted media. Oxygen evolution rates for the mutant were further depressed (28% of control rates) under chloride-limiting conditions. Addition of bromide restored these rates to those observed under chloride-sufficient conditions. Measurements of the variable fluorescence yield indicated that the mutant assembled fewer functional centers in the absence of chloride. These results indicate that the mutation K321G in CP 47 affects PSII stability and/or assembly under conditions where chloride is limiting. PMID- 9225858 TI - Expression of arginine decarboxylase in seedlings of indica rice (Oryza sativa L.) cultivars as affected by salinity stress. AB - The effect of salinity stress on the activity of arginine decarboxylase (ADC, EC 4.1.1.19), the first enzyme in biosynthesis of polyamines (PA) from arginine, as well as its transcript level has been compared in salt-sensitive (M-1-48) and salt-tolerant (Pokkali) rice cultivars. Treatment of 72 h grown seedlings either with increasing concentrations of NaCl or with 150 mM NaCl for different time periods, showed a gradual increase of activity in Pokkali. In M-1-48 an immediate increase followed by sharp decrease was observed on prolonged treatment beyond 6 h or above 150 mM NaCl. To generate a DNA probe for ADC, the polymerase chain reaction was used with oat genomic DNA and sequence-specific primers. A region of oat genomic DNA containing a coding sequence for 166 amino acids of the C terminal part of the ADC enzyme was amplified and called OAD1. Southern analysis of EcoRI- or BamHI-cut genomic DNAs from different cultivars of rice with OAD1 as the probe revealed strong hybridization with one DNA fragment of rice and restriction fragment length polymorphism (RFLP) was noticed. Northern analysis of total RNA of rice with OAD1 as the probe revealed hybridization with a transcript of similar size to the ADC transcript in oat. While in Pokkali, at least a 20 fold accumulation of OAD1 homologous transcript was detected after treatment with 200 mM NaCl, only a seven-fold increase in transcript level was found in M-1-48 after 150 mM NaCl treatment. Results suggest that in the salt-tolerant rice cultivar Pokkali, ADC enzyme activity increases and its transcript also accumulates during the prolonged salinity stress, this mechanism is absent in the salt-sensitive rice cultivar M-1-48 where a prolonged period of salinity stress down-regulates both ADC activity and its transcript level. PMID- 9225859 TI - Specific sequence modifications of a cry3B endotoxin gene result in high levels of expression and insect resistance. AB - Solanum melongena (eggplant) cv. Picentia and the wild species Solanum integrifolium were transformed with both a wild type (wt) and four mutagenized versions of Bacillus thuringiensis (Bt) gene Bt43 belonging to the cry3 class. The Bt gene was partly modified in its nucleotide sequence by replacing four target regions (W: +1 to +170; X: +592 to +1057; Y: +1203 to +1376; Z: +1376 to +1984) with synthetic fragments obtained by polymerase chain reaction amplification of crude oligonucleotides. The synthetic Bt genes were designed to avoid, in their modified regions, sequences such as ATTTA sequence, polyadenylation sequences and splicing sites, which might destabilize the messenger RNA. Furthermore, the codon usage was improved for a better expression in the plant system. The amino acid composition was not altered. Four versions of the modified Bt gene were obtained, BtE, BtF, BtH and BtI, with a nucleotide subtitution percentage of 8.2, 8.6, 14, and 16%, respectively, in comparison to the wt gene Bt43. Modified versions contained different subsets of substituted regions: BtE-W + Z, BtF - Y + Z, BtH-X + Y + Z, BtI - W + X + Y + Z. In the final modified version (BtI), overall guanine+cytosine was increased from the 34.1% of the wt gene to 45.5%, and most of the destabilizing sequences were eliminated. Transgenic plants obtained with the more modified versions, BtH and BtI, were fully resistant to Leptinotarsa decemlineata Say first- and third-instar larvae, while Bt43 wt, BtE and BtF genotypes did not cause mortality and did not affect larval development. PMID- 9225860 TI - Isolation of a cDNA encoding an Arabidopsis galactokinase by functional expression in yeast. AB - A cDNA clone encoding Arabidopsis thaliana galactokinase was fortuitously isolated during the course of a screen for plant homologues of a Saccharomyces cerevisiae peroxisome assembly gene, PAS9. Clones were sought which restored the ability of pas9 cells to grow on oleate as a sole carbon source, as oleate metabolism requires peroxisomal beta-oxidation and therefore functional peroxisomes. Subsequent experiments showed that high level expression of the galactokinase cDNA did not complement the peroxisomal assembly defect, but instead permitted the cells to grow on agar plates in the absence of an external carbon source. Agar plates were shown to contain a small amount of galactose released from the agar as a result of autoclaving. The galactokinase clone was shown to be functional, as it could complement a S. cerevisiae galactokinase mutant. Galactokinase is a single copy gene in Arabidopsis, which has been designated AGK1, and is expressed in all the major organs of the plant. PMID- 9225861 TI - Interaction of rice and human SRP19 polypeptides with signal recognition particle RNA. AB - The signal recognition particle (SRP) controls the transport of secretory proteins into and across lipid bilayers. SRP-like ribonucleoprotein complexes exist in all organisms, including plants. We characterized the rice SRP RNA and its primary RNA binding protein, SRP19. The secondary structure of the rice SRP RNA was similar to that found in other eukaryotes; however, as in other plant SRP RNAs, a GUUUCA hexamer sequence replaced the highly conserved GNRA tetranucleotide loop motif at the apex of helix 8. The small domain of the rice SRP RNA was reduced considerably. Structurally, rice SRP19 lacked two small region that can be present in other SRP19 homologues. Conservative structure prediction and site-directed mutagenesis of rice and human SRP19 polypeptides indicated that binding to the SRP RNAs occurred via a loop that is present in the N-domain of both proteins. Rice SRP19 protein was able to form a stable complex with the rice SRP RNA in vitro. Furthermore, heterologous ribonucleoprotein complexes with components of the human SRP were assembled, thus confirming a high degree of structural and functional conservation between plant and mammalian SRP components. PMID- 9225862 TI - Differential display-mediated isolation of a genomic sequence for a putative mitochondrial LMW HSP specifically expressed in condition of induced thermotolerance in Arabidopsis thaliana (L.) heynh. AB - Plants of Arabidopsis thaliana pre-treated at 37 degrees C for 2 h can survive an otherwise lethal heat shock at 45 degrees C. Differential display reverse transcriptase-PCR (DDRT-PCR) was utilized to clone DNA fragments corresponding to mRNAs specifically expressed in conditions of induced thermotolerance or of expression of thermotolerance. One of these DDRT-PCR fragments enabled the isolation of a genomic clone pAt1.3EX, containing the sequence Athsp23.5, the gene for a low-molecular-weight (LMW) heat shock protein (HSP), AtHSP23.5. Athsp23.5 is low- or single-copy in the Arabidopsis genome and its open reading frame is interrupted by a 137 bp intron. Analysis of the sequence suggests AtHSP23.5 is targeted to the mitochondrion. The steady-state level of the AtHSP23.5 mRNA varied significantly according to the heat treatment, increasing on heat shock (transfer from 22 degrees C to 37 degrees C), with a further increase during expression of thermotolerance (transfer from 22 degrees C to 37 degrees C and then to 45 degrees C). Expression was low after an abrupt stress (from 22 degrees C to 45 degrees C). This behaviour was different from that observed for other LMW HSP mRNAs that were present at high level at 37 degrees C, but did not increase significantly in condition of expression of thermotolerance, and reached a considerable steady-state level also during the abrupt stress at 45 degrees C. The retrotranscription of AtHSP23.5 mRNA followed by amplification with two primers encompassing the intron allowed for the isolation of an almost full-length cDNA sequence. The sequence analysis of the two cDNAs obtained from condition 22 degrees C-->37 degrees C and condition 22 degrees C-->45 degrees C suggested that in both cases the intron had been correctly spliced. The importance of correct intron splicing in survival at high temperatures and the role of mitochondrial HSP in induction and expression of thermotolerance are discussed. PMID- 9225863 TI - Chromosomal location and expression of the single-copy gene encoding high mobility-group protein HMG-I/Y in Arabidopsis thaliana. AB - A cDNA encoding the HMG-I/Y protein from Arabidopsis thaliana has been isolated and characterised by nucleotide sequencing. The 903 bp cDNA contains a 612 bp open reading frame encoding a protein of 204 amino acid residues showing homology to HMG-I/Y proteins from other plant species. The protein contains four copies of the 'AT-hook' motif which is involved in binding A/T-rich DNA. Southern blotting showed that the HMG-I/Y gene was present in a single copy in the Arabidopsis genome. The gene was localised to the top of chromosome 1 by RFLP analysis of F8 recombinant inbred lines. Northern blotting showed that the gene was expressed in all organs examined, with the highest expression in flowers and developing siliques. PMID- 9225864 TI - The Brassica rapa elongated internode (EIN) gene encodes phytochrome B. AB - The elongated internode (ein) mutation of Brassica rapa leads to a deficiency in immunochemically detectable phytochrome B. Molecular analysis of the PHYB gene from ein indicates a deletion in the flanking DNA 5' of the ATG start codon, which could interfere either with PHYB transcription or processing of the PHYB transcript. Restriction fragment length polymorphisms and inverse PCR fragments generated from the PHYB gene of wild-type and ein seedlings demonstrate the deletion to be 500 bp in length. Seedlings of heterozygote, EIN/ein, contain about 50% of the level of immunochemically detectable phytochrome B of equivalent wild-type EIN/EIN seedlings. Etiolated seedlings of EIN/ein show a responsiveness to red light almost intermediate between that of ein/ein and EIN/EIN homozygotes. Furthermore, whereas the ein/ein homozygote is poorly responsive to low red/far red ratio light, the presence of one functional allele of EIN in the heterozygote confers an elongation response intermediate between that of the homozygotes EIN/EIN and ein/ein in these light conditions. The partial dominance of ein indicates a close relationship between phytochrome B level and phenotype. PMID- 9225865 TI - Promoter sequences from two different Brassica napus tapetal oleosin-like genes direct tapetal expression of beta-glucuronidase in transgenic Brassica plants. AB - To investigate the sequences responsible for the regulated expression of tapetal specific oleosin-like genes, ca. 2 kb of the 5'-upstream regions from two divergent genes, OlnB;4 and OlnB;13, were isolated, sequenced and fused to the reporter gene beta-glucuronidase for study in transgenic Brassica napus plants. Although the proteins encoded by these two genes are highly divergent, except for the conserved oleosin-like domain, the first 250 bp of their 5'-upstream regions was 86% identical, including a region of 150 bp upstream from the TATA box. Analysis of 42 independent transformants by histochemical and fluorometric methods showed that both promoters directed tapetal-specific expression that peaked at the 4 mm flower bud stage. PMID- 9225867 TI - Activation of endothelial cells in thrombosis and vasculitis. AB - The endothelium participates actively in homeostatic mechanisms such as the regulation of vascular tone and maintenance of a nonthrombotic environment, as well as directing biological responses such as leukocyte trafficking to inflammatory sites. Disruption of these processes leads to disease. In the antiphospholipid antibody syndrome autoantibodies provoke the endothelium to develop a prothrombotic surface. In systemic vasculitides associated with presence of antineutrophil cytoplasm antibodies, it is likely that the autoantibodies incite premature neutrophil activation, disrupted neutrophil endothelium interactions and endothelial damage. This review considers how normal endothelial functions may be subverted in disease and how active endothelial responses may contribute to disease. PMID- 9225866 TI - Characterization and expression of a rice RAD23 gene. AB - In order to identify proteins that interact with plant transcriptional complexes, we performed a two-hybrid screen in yeast using a cDNA library from embryogenic rice suspension cultures and the plant transcriptional activator viviparous-1 (vp1) as 'bait'. In this screen, we detected an interaction between VP1 and a rice homologue of the Saccharomyces cerevisiae RAD23 gene (osRAD23). The RAD23 protein is associated with the general transcriptional machinery in yeast, and is believed to play a role in the processes of nucleotide excision repair in yeast and mammalian cells. This report is the first identification of a RAD23 homolog in plants. The osRAD23 amino acid sequence shares 50-60% similarity throughout its length with RAD23 sequences from yeast, mice, and man. osRAD23 contains a characteristic ubiquitin-like domain at its N-terminus, which is similar to other RAD23 genes. Analysis of the expressed sequence tag database identifies two different classes of RAD23 genes in both Arabidopsis and rice. Southern analysis of rice genomic DNA indicated the presence of at least two RAD23-like genes. A single transcript (1.5 kb) of osRAD23 was detected in total RNA from rice embryonic tissue, while three transcripts (1.8, 1.5 and 1.0 kb) were observed in total RNA from vegetative tissues of rice. PMID- 9225868 TI - Orchitis due to vasculitis in autoimmune diseases. AB - Autoimmune diseases can affect the blood vessels, causing systemic vasculitis. Although testicular manifestation of some autoimmune diseases is not uncommon, only a few cases of acute orchitis are described in the literature. The underlying pathological condition in testicular manifestations of autoimmune diseases is severe vasculitis causing inflammation and infarction. In patients with recurrent episodes of scrotal swelling and pain, testicular vasculitis as the first sign of a systemic disease should be taken into consideration. PMID- 9225870 TI - T cell receptor (V beta) bias in the response of rheumatoid arthritis synovial fluid T cells to connective tissue antigens. AB - T cell receptor (V beta) use in the response to type II collagen and cartilage proteoglycans was analysed in peripheral blood and synovial fluid T cells from RA patients. T cells from RA patients with an immune response to connective tissue antigens, and paired PB and SF samples were stimulated in vitro with type II collagen, high density aggrecan proteoglycans (PG), and the T cell mitogen concanavalin A. After short term culture, mRNA was extracted from cells and a reverse transcription-polymerase chain reaction was performed, using primers specific for eight TCR V beta determinants. Blood cells stimulated with ConA generated strong bands with virtually all the V beta primers tested, but the TCR (V beta) expression by SF T cells stimulated with mitogen was biased, suggesting a selection process during joint infiltration. The V beta phenotypes of cells responding to PG was restricted in individual RA patients, but the pattern of V beta use in the the RA population was not consistent. In contrast, the V beta phenotypes of SF cells responding to CII was highly biased in both individual patients and the RA population, with V beta 14, V beta 17, and V beta 8 phenotypes predominant. We conclude that the T cell response to connective tissue antigens is restricted compared with mitogen stimulation, with the highest degree of TCR bias seen in the response of SF T cells to stimulation with type II collagen. PMID- 9225869 TI - Reliability of radiographic grading of osteoarthritis of the hip and knee. AB - We review studies on the reliability of radiographic assessment of osteoarthritis of the hip and knee. Reliability studies were reported for 10 among 24 identified scores. In general, moderate to good agreement was found for overall scores and for separate grading of joint space narrowing of the hip and osteophytes of the knee in the majority of studies, while reliability tended to be lower for other radiographic features. Overall scores of the knee were more reliable than overall scores of the hip, and intra-rater-reliability was considerably higher than inter rater-reliability in most instances. Comparison of reliability between scores can only be made with caution, given the difference in the design of reliability studies, particularly the different qualification of involved observers. The limits of existing knowledge on reliability of commonly used radiologic scores are outlined, and, proposals are made to overcome those limits in future studies. PMID- 9225871 TI - Association of DM genes in systemic sclerosis is secondary to the association with HLA genes. AB - The contribution of polymorphism of DMA and DMB alleles to the pathogenesis of Japanese Systemic Sclerosis (SSc) was studied in 55 Japanese SSc patients and 77 normal Japanese subjects using the PCR-RFLP (restriction fragment length polymorphism) method. The allele frequencies of DMB*0101 allele were increased in SSc with diffuse scleroderma (70.0% vs 49.4%, p < 0.05, pc = not significant (NS)) and in SSc with antitopoisomerase I antibody (a-Scl-70), (68.2%, p < 0.05, pc = NS). The phenotype frequencies of DMB*0101 in these subgroups of SSc were increased significantly (95.0%, p = 0.014, pc < 0.05; 95.5%, p = 0.0088, pc < 0.05, respectively). In contrast, DMB*0102 and DMB*0103 alleles tended to decrease in diffuse scleroderma and SSc with a-Scl-70, but the decreases were not significant. Association analysis among DMA, DMB, and DRB1*1502 in Japanese SSc with diffuse scleroderma and SSc with a-Scl-70 indicated that the increase in DMA*0101 was not primary, but reflected an increase in HLA DRB1*1502. PMID- 9225872 TI - Non-Hodgkin's lymphomas complicating Sjogren's syndrome: can Epstein Barr virus be implicated? AB - We examined eight (6 parotid, 1 caecum, 1 lymph node) non Hodgkin's Lymphomas (NHL) complicating primary Sjogren's syndrome (SS), four parotid NHL, in patients without SS, and three salivary gland biopsies from SS patients and no NHL, for Epstein Barr virus (EBV), using immunohistochemistry for late membrane protein, in situ hybridisation (ISH) for EBER, and PCR for EBV DNA. Late membrane protein was not detected. In NHL's complicating SS, EBERs were detected in two parotid lymphomas by ISH. EBV DNA was detected in three SS parotid NHL. Cecal and lymph node SS NHL were negative for EBER and EBV DNA. EBV DNA was detected in two non SS NHLs, one expressed EBER. Despite positive EBV DNA results by PCR in three samples expression of EBER was noted in only one by ISH. This was a high grade NHL complicating SS. There was no evidence of EBV in low grade NHLs complicating SS. PMID- 9225873 TI - Avascular necrosis of bone in systemic lupus erythematosus. The predictive role of precipitating autoantibodies. AB - The association between the type of precipitating autoantibodies and occurrence of avascular necrosis of bone (AVN) was examined. We prospectively analyzed clinical and laboratory findings of our 113 patients with SLE. Seven of 113 (6%) patients developed AVN. Anti-Ro (SS-A) and anti-RNP antibodies coexisted in 3 of 7 AVN patients. The same combination of these two antibodies were observed in 1 without AVN. Antibodies to topoisomerase I were detected in 2 other patients with AVN but not in any of the patients without AVN. The coexistence of the former two or the presence of the latter one is rare in SLE. However, these (combination of) antibodies can be useful as a local ischemic marker predicting the development of AVN. PMID- 9225874 TI - Correlates of disablement in systemic onset juvenile chronic arthritis. A cross sectional study. AB - The impact of systemic onset JCA on functional outcome was studied in a multidimensional construct. Twenty-one patients were subjected to auxologic evaluation, a laboratory check, pulmonary and cardiac function tests, radiographic evaluation, joint count on tenderness, swelling and function, ADL, health assessment (CHAQ), and psychosocial evaluation. Six of 21 patients had active systemic disease. Restrictive pulmonary function was found in 8/17 patients, 1/21 had pericarditis. Joint impairment was moderate. Functional limitations were mild. Self-esteem was positive. Parental report on functional limitation correlated significantly with joint impairment. Performance of daily activities correlated strongly with perceived competence. Active inflammatory disease did not correlate with joint impairment and functional limitation. Patients with systemic onset JCA develop mild functional limitations, which partially correlate with the more serious impairments. Pulmonary function disorders are a common impairment. Active inflammatory disease might influence functional outcome, but there is no evidence that it influences joint impairment outcome. PMID- 9225875 TI - Quantitative assessment of clinical disease status in primary Sjogren's syndrome. A cross-sectional study using a new classification model. AB - Quantitative and qualitative assessment of the clinical disease manifestations in 41 primary Sjogren's syndrome (pSS) patients was performed according to a new classification model. Frequencies of subgrouped disease manifestations were as follows: 1) surface exocrine disease: 100%, 2) internal organ exocrine disease: 63%, 3) monoclonal B lymphocyte disease: 5%, 4) inflammatory vascular disease: 71%, 5) non-inflammatory vascular disease: 59%, 6) mediator induced disease: 98%. Summary scores for severity of surface exocrine disease correlated to the summary scores of all other disease manifestations (p = 0.02), to the summary scores of internal organ exocrine disease (p = 0.003), and to the summary scores of mediator induced disease (p = 0.03). Blood leucocyte counts showed significant negative correlations to levels of plasma IgG, serum IgA-RF, IgM-RF, anti-SSA/SSB antibodies, IL-6, and IL-1Ra. We conclude that the model made detailed analysis of the clinical presentation of pSS possible, and thus may assist in elucidating important pathobiological aspect of the disease. PMID- 9225876 TI - Double-blind, placebo-controlled cross-over study of intravenous S-adenosyl-L methionine in patients with fibromyalgia. AB - The objective of this study was to test the efficacy of intravenously administered S-adenosyl-L-methionine (SAMe) in patients with fibromyalgia (FM). Thirty-four out-patients with fibromyalgia symptoms received SAMe 600 mg i.v. or placebo daily for 10 days in a cross-over trial. There was no significant difference in improvement in the primary outcome: tender point change between the two treatment groups. There was a tendency towards statistical significance in favour of SAMe on subjective perception of pain at rest (p = 0.08), pain on movement (p = 0.11), and overall well-being (p = 0.17) and slight improvement only on fatigue, quality of sleep, morning stiffness, and on the Fibromyalgia Impact Questionnaire for pain. No effect could be observed on isokinetic muscle strength, Zerrsen self-assessment questionnaire, and the face scale. No effect of SAMe in patients with FM was found in this short term study. PMID- 9225877 TI - Ultrasound guided synovial biopsy of the wrist. AB - Seven patients (4 female and 3 male, mean age 46) with arthritis of the wrist (n = 7) without known etiology were evaluated. High-definition ultrasound equipment was used for localization of synovial hypertrophy, suitable for ultrasound guided biopsy without risk. A 18-gauge diameter Tru-cut biopsy needle was used in an automated gun. Two passes were performed with continuous guidance of the needle tip. In all patients representative synovial tissue was obtained in adequate amount. No complications were encountered. Ultrasound guided biopsy is proposed as an effective technique which can be performed with low patient discomfort on an outpatient basis. PMID- 9225878 TI - Nodular regenerative hyperplasia of the liver in systemic lupus erythematosus. The relationship with anticardiolipin antibody and lupus anticoagulant. AB - Recent reports have indicated that nodular regenerative hyperplasia (NRH) associated with systemic lupus erythematosus (SLE) is related to anticardiolipin antibodies (aCL) and/or lupus anticoagulant (LA). We describe a patient with SLE complicated by NRH, who did not show neither aCL nor LA activity. This case suggests that the pathogenesis of NRH in patients with autoimmune diseases is heterogeneous and not confined to aCL and LA. PMID- 9225880 TI - Carpal tunnel syndrome heralding polymyalgia rheumatica. AB - Presenting symptoms in polymyalgia rheumatica (PMR) may be atypical. We report herein two old females who developed a bilateral carpal tunnel syndrome several months before the typical symptoms of PMR appeared. In both patients the diagnosis of PMR was overlooked and a surgical release of the median nerve was performed. PMR should be considered in elderly people who develop an acute or subacute carpal tunnel syndrome. PMID- 9225879 TI - Aneurysmal dilatation of ascending aorta and aortic insufficiency in juvenile spondyloarthropathy. AB - Aortic insufficiency is a well-recognized complication of adult spondyloarthropathy but rare in juveniles, and no report is associated with aneurysmal dilatation at any age. We describe rapidly progressive aortic insufficiency with aneurysmal dilatation of ascending aorta in a 15-year-old boy with juvenile spondyloarthropathy, necessitating a Bentall operation. PMID- 9225881 TI - Multiorgan sarcoidosis presenting with symmetric polyarthralgia, cutaneous vasculitis, and sicca symptoms. AB - A 49 year old male presented with symmetric polyarthralgia, cutaneous vasculitis, and sicca symptoms. Histological examinations of biopsy specimens from the liver and skin showed sarcoidosis. PMID- 9225882 TI - Thoracic myelopathy due to intraspinal rheumatoid nodules. AB - A fifty-six-year-old woman with classical rheumatoid arthritis had subacute onset of paraparesis due to thoracic epidural rheumatoid nodules. Although plain radiograms and computed tomograms of the thoracic spine were negative except for old compression fractures, magnetic resonance imaging revealed thoracic spinal cord compression due to masses at multiple levels. There was a steady recovery after excision surgery. PMID- 9225883 TI - Classification criteria for rheumatoid arthritis. PMID- 9225884 TI - Hazards associated with metalworking by artists. AB - Metalworking is the constructive sculptural application of various technologic processes for producing art from ferrous and nonferrous metals. Self-employed artists and their workplaces are not protected by governmental agencies. We describe the techniques involved in metalworking processes and discuss the physical trauma and medical syndromes associated with each process, as well as safety, prevention, and treatment aspects of the basic syndromes. PMID- 9225885 TI - Intracranial saccular (berry) aneurysm: a brief overview. AB - An incidental saccular intracranial (berry) aneurysm is not an uncommon finding at autopsy. Rupture of such an aneurysm can produce severe neurologic deterioration and death, often without warning. Morbidity and mortality can be reduced in some patients with early surgical intervention. Familial aggregation of cerebral aneurysms occurs frequently enough to warrant screening of family members to diagnose an asymptomatic berry aneurysm. We review the etiology, clinicopathologic features, and natural history of berry aneurysms and aneurysm rupture, as well as the various screening methods. A careful examination of the cerebral vessels should be routine in every postmortem examination, regardless of the cause of death. PMID- 9225886 TI - Blood glucose determination: point of care testing. AB - Mechanisms for point of care glucose determinations have changed significantly since first introduced approximately 20 years ago. Such tests are now commonly done in acute and chronic care hospitals, as well as in physicians' offices and patients' homes. Although basically reliable, there are a number of potential problems with glucose determination by these methods that may not be considered by physicians interpreting these tests. This is a brief review of such problems, especially in the acute care setting. PMID- 9225887 TI - The addition of antibiotics to fibrin glue. AB - Fibrin glue is composed of two separate solutions of fibrinogen and thrombin. When mixed together, these two solutions mimic the final stages of the clotting cascade to form a fibrin clot. Because the resulting fibrin patch is a good medium for microbial growth, the addition of antibiotics to one of the components of fibrin glue has been shown to reduce postoperative infections. Seventeen different antibiotics have been investigated in vitro. Of the 17, cefotaxime, mezlocillin, gentamicin, neomycin, and polymixin B, when added to fibrin glue, can decrease the rate of clot formation or the strength of the resultant fibrin clot. Further work is necessary to characterize the effect the addition of antibiotics has on the rate and strength of fibrin clotting and to determine what effect low systemic levels of antibiotics might have on antibiotic resistance patterns. PMID- 9225888 TI - Physician education in hyperlipidemia management: the impact on collaboration. AB - Collaboration of health care professionals is likely beneficial in modifying patient behavior in the treatment of hyperlipidemia. The purpose of this study was to determine whether limited instruction and demonstration of collaborative management of hyperlipidemia in a continuing medical education (CME) would change physicians' office practices, as determined 1 year later by questionnaire. Collaborative practice was defined as physicians working with other allied health care professionals as a team to increase patients' medication compliance and other behavioral outcomes. A 19-credit hour CME Lipid Disorders Training Program (LDTP) was offered emphasizing the collaborative approach to hyperlipidemia patient management. Physicians (n = 196) were surveyed 1 year after LDTP. The response rate was 52.5%, nonrespondents were similar in locations. About 51% of respondents reported increased collaborative practice; of these respondents, 68% reported saving time, 78% reported improved patient outcomes, 76% improved office efficiency, and 90% increased patient satisfaction. According to self-reporting by these physicians, increased collaboration practices after attending the LDTP course led to improved patient outcomes. PMID- 9225889 TI - Sertindole hydrochloride: a novel antipsychotic medication with a favorable side effect profile. AB - Forty percent of all long-term care hospitalization days are accounted for by patients with schizophrenia. New approaches to managing this disorder are needed, including improved efficacy and better tolerability to enhance compliance with treatment. Sertindole hydrochloride is a novel antipsychotic medication soon to be available in the United States and Canada. As part of multisite phase II and III studies, we studied effects of this medication in five patients with chronic schizophrenia and examined the side effect profile. With more than 30 patient months of exposure, sertindole treatment was not associated with neurologic side effects and was well tolerated in all patients studied. No evidence of hematologic abnormalities was found. Serial electrocardiograms revealed slight increases in QTc that were not considered clinically significant and did not lead to discontinuance of treatment. While data from larger samples are needed, in this small population sertindol hydrochloride was tolerated well with no evidence of acute neurologic side effects associated with traditional treatments for schizophrenia. Individuals with schizophrenia may benefit from enhanced compliance with treatment and a possible reduction in hospitalizations in the future. PMID- 9225890 TI - Ender nails: an alternative for intramedullary fixation of femoral shaft fractures in children and adolescents. AB - Fixation of femur fractures in children and adolescents is becoming widely accepted because of the lower chance of iatrogenic infection and prohibitive cost of in-hospital traction and spica cast care. Interlocking nails, which have dramatically improved adult femur fracture care, have posed problems, such as injury to the greater trochanteric apophysis and osteonecrosis of the femoral head. We reviewed a series of five stable femoral shaft fractures treated with Ender nails in children from 9 to 17 years of age. No serious complications (eg, leg length inequality) occurred. Follow-up averaged more than 3 years. Complete healing, with return to preinjury activity levels, occurred within 11 weeks on average. Decreased hospitalization, low cost of implants, less potential damage to growth centers, and decreased blood loss and operative time suggest this procedure has merit. Attention to operative technique diminishes previously reported complications attributed to Ender nails. PMID- 9225891 TI - Acquired immunodeficiency syndrome in Alabama: special concerns for black women. AB - BACKGROUND: Among AIDS case reports from rural and small town areas of the United States, rates are higher in the South than in any other part of the country. METHODS: For this study, we analyzed AIDS surveillance statistics from the state of Alabama for trends, distributions, and populations affected. We aggregated Alabama AIDS surveillance data in 5-year intervals--1981 to 1985, 1986 to 1990, 1991 to 1995--and made comparisons based on geographic area of residence of people diagnosed with AIDS. RESULTS: Of the 3,558 cases of AIDS reported in Alabama in the period 1981 to 1995, 86% were men and 14% were women. Among women, 69.7% were black and 29.1% were white. Among men, 48.4% were black and 50.9% were white. We compared these figures with 1995 Alabama population estimates of 26.2% black, 73.7% white, and < 1% another race. The rates for black women and white women increased 170-fold and 23-fold, respectively, from the 1981 to 1985 period to the 1991 to 1995 period. For the same periods, case rates for black men and white men increased more than 80-fold and 50-fold, respectively. Black women showed a rise per 100,000 population-from 0.3 (1981 to 1985) in both northern and southern Alabama to 37 in northern Alabama and 64 in southern Alabama (1991 to 1995). CONCLUSIONS: Black women are at disproportionately high risk, particularly in the southern counties of Alabama. HIV is increasingly prevalent in rural and small town communities in Alabama and is more often transmitted heterosexually than it has been previously. PMID- 9225892 TI - Unexpected second trimester pregnancy loss due to maternal parvovirus B19 infection. AB - Five unexplained second trimester fetal deaths occurring in a 3-month interval were evaluated to determine whether the fetal demise was associated with maternal infection due to parvovirus B19. One fetus was markedly hydropic, a classic finding of fetal infection; no gross anomalies were present in the other four. Liver specimens from all five fetuses showed viral inclusions by light microscopy, supporting the presence of fetal parvovirus B19 infection. None of the five women had a history of parvovirus infection, but serum samples from all of them were positive for IgG to parvovirus. We conclude that maternal parvovirus B19 infection is associated with fetal demise during the second trimester and that hydropic changes may be absent, necessitating careful evaluation of tissue from the dead fetus. PMID- 9225893 TI - Endoscopic retrograde cholangiopancreatography in the treatment of bile leaks and bile duct strictures after laparoscopic cholecystectomy. AB - We reviewed our experience over 3 years with 11 patients who had bile leaks (Group 1) and 8 patients who had bile duct strictures after laparoscopic cholecystectomy (LC) and were treated with endoscopic retrograde cholangiopancreatography (ERCP) (Group 2). In Group 1, bile leaks were at the level of the cystic duct in 10 patients and from a duct of Luschka in 1 patient; 10 patients had sphincterotomy and 11 patients received barbed stents. All patients had resolution of bile leak and stents were removed after an average of 5 weeks. In Group 2, stenoses were at the level of the common bile duct (CBD) in 7 patients and of the CBD-common hepatic duct in 1 patient. Six patients had a sphincterotomy and 7 patients were successfully treated with pneumatic polyethylene balloon dilatation and stent placement. One patient had unsatisfactory dilatation and was referred to surgery. Two patients had permanent resolution of stenosis at 3 and 4 years of follow-up, 5 patients had recurrence, and a total of 6 patients eventually needed surgery. We conclude that ERCP is effective in resolving isolated bile leaks, but iatrogenic strictures after LC more often require surgical treatment after ERCP. PMID- 9225894 TI - Peer discussion on training physicians to be competent communicators: support for a multiple discourse approach. AB - To consider medical educators' views about training physicians to be competent communicators, a focus group was conducted at a large southeastern medical college with faculty responsible for training interns and residents to be competent communicators. The medical school was undergoing a curriculum review and instructional development process, including an examination of the methods being used to teach communication. A discussion of a teaching faculty committee revealed several themes, including the foundational assumption that communication is a natural ability that some have and others do not. Other notions likely to affect the educators' beliefs about the importance and effectiveness of communication education include the ideas that courtesy is synonymous with good communication and that although a few fundamentals might be transmitted, experience is the best teacher. Participants identified several audiences, in addition to patients, with whom they communicate as part of their profession, including other physicians, other health care providers (e.g., nurses and lab technicians), and hospital administrators, health care insurers, and others responsible for evaluating physicians' performance. The implications of this project for the development of medical communication courses within a multiple discourse approach are discussed. PMID- 9225895 TI - Diagnosis of Alzheimer's disease in a community hospital-based brain bank program. AB - Several studies in recent years have addressed the accuracy of the clinical diagnosis of Alzheimer's disease (AD). However, most large studies have been done in university centers specializing in dementia. The purpose of this study was to assess the diagnostic accuracy of dementia in a community-based brain bank program, using data from the Central Florida component of the State of Florida's Brain Bank Program. Since 1987, 261 cases of dementia have been assessed at antopsy, and the clinical and pathologic diagnoses were compared. Of 234 patients with a clinical diagnosis of AD, 181 (77%) had a pathologic diagnosis of AD, with or without other contributing disorders. Of 27 patients with a clinical diagnosis of non-AD dementia, 14 (52%) had a pathologic diagnosis of AD, with or without other contributing diagnoses. These findings are similar to those previously reported and emphasize the importance of autopsy for the accurate diagnosis of dementia for genetic counseling, assessment of diagnostic techniques or drug therapy, and epidemiologic studies. PMID- 9225896 TI - Surgical treatment of gastrointestinal B-cell mucosa-associated lymphoid tissue lymphomas. AB - We retrospectively evaluated clinicopathologic features of 35 patients treated for primary gastrointestinal lymphomas of MALT type (mucosa-associated lymphoid tissue) between 1970 and 1993. Fourteen patients (40%) were treated for acute abdominal conditions (bowel obstruction in 8, perforation in 2, and gastrointestinal bleeding in 4), and the rest had exploratory laparotomy. The tumor was located in the stomach in 23 patients (66%), in the jejunum and ileum in 10 (29%), and in the large intestine in 2 (6%). The type of operation was defined according to site and extent of disease. Most patients received chemotherapy postoperatively. Staging was done according to the Ann Arbor classification. Survival depended on stage and extension of the disease; 5-year survival was 45%. Surgical resection followed by adjuvant chemotherapy is warranted when the patient is considered to be a surgical candidate. PMID- 9225897 TI - Synchronous pulmonary cryptococcosis and histoplasmosis. AB - A chronically immunosuppressed patient receiving oral steroid therapy had a cavitary lesion; both Histoplasma capsulatum and Cryptococcus neoformans were found in samples of respiratory secretions. C neoformans was also found in specimens taken from skin lesions and blood. This is the first reported instance of synchronous infection by these two fungi. PMID- 9225898 TI - En bloc pancreaticoduodenectomy and colectomy for duodenal neoplasms. AB - Duodenal malignancy is rare and generally considered to have both a low resectability rate and a poor prognosis. Historically, the involvement of the colon or its mesentery has been considered a criterion for unresectability by many surgeons because of the overall magnitude of surgery involved with an en bloc colectomy and pancreaticoduodenectomy. In the past few years, several reports have noted a decrease in morbidity and mortality rates for pancreaticoduodenectomy. The current safety of the procedure suggests that the classical criteria for resectability can now be reevaluated for certain neoplasms. We report two cases of pancreaticoduodenectomy with en bloc colectomy done as attempted curative resections for primary duodenal malignancies. The procedure was well tolerated by both patients; there were no major complications, and it provided both prolonged survival and effective palliation. PMID- 9225899 TI - Hemangioma of the temporal bone in a patient presumed to have Meniere's syndrome. AB - This case report describes a patient with a facial nerve hemangioma of 8 years' duration that initially caused most of the symptoms of Meniere's syndrome: fullness, sensorineural hearing loss, dizziness, tinnitus, and disruption of balance. The hearing loss was in the high-frequency range (> or = 3,000 Hz); typically, the initial hearing loss in Meniere's syndrome is in the low-frequency range. Mild facial nerve weakness and punctate keratitis due to corneal exposure appeared 8 years later. Contrast-enhanced magnetic resonance imaging and high resolution computed tomography depicted the lesion and made preoperative diagnosis possible. With meticulous surgical removal of the tumor, which was intertwined with the facial nerve, facial nerve function was preserved. PMID- 9225900 TI - Cardiac rupture due to blunt trauma. AB - Cardiac rupture due to blunt trauma has been recognized with increasing frequency over the past two decades. The mortality rate is high and the majority of patients die before they arrive at the emergency department. A high index of suspicion and prompt surgical intervention are crucial for survival. We report the management of two patients, one with a double right atrial tear and one with a single right atrial tear, after each was involved in a motor vehicle accident. PMID- 9225901 TI - Intravascular tumor: a previously unreported finding of glucagonoma. AB - Glucagonoma is a relatively rare pancreatic islet cell tumor. Historically, these tumors present a typical constellation of symptoms including diabetes, weight loss, anemia, necrolytic migratory erythematous rash, and propensity for thrombosis. This clinical presentation is described as the glucagonoma syndrome. The syndrome can be confirmed with the use of serum measurements of glucagon levels and immunohistochemical assay of the tumor. Variations from the classic syndrome have been described, and serum measurements of glucagon in patients with suspected islet cell tumors can identify subsets of patients with glucagonoma who do not exhibit the classic syndrome. In our case, the unusual presentation of glucagonoma included the previously unreported component of an intravascular venous extension of tumor. PMID- 9225902 TI - Primary subclavian vein thrombosis and pulmonary embolism. AB - Thrombotic veno-occlusion of the upper extremity is infrequent in the absence of external trauma or in the presence of an indwelling catheter. Inadvertent injury to the underlying venous circulation can occur when the upper extremity is subjected to an unaccustomed repetitive action. The resultant intimal damage may become a nidus for thrombus development. PMID- 9225903 TI - Cellulitis and myositis caused by Agrobacterium radiobacter and Haemophilus parainfluenzae after influenza virus vaccination. AB - Agrobacterium radiobacter is a gram-negative aerobic bacillus that has been reported as a cause of disease only 36 times in the literature. More than half of the patients (25) have had bacteremia. Peritonitis, urinary tract infection, endocarditis, and one case of cellulitis associated with bacteremia have also been reported. Infection is often associated with immunosuppression and the presence of a plastic foreign body, such as central venous catheters, nephrostomy tubes, intraperitoneal catheters, and prosthetic cardiac valves. We present apparently the first case of A radiobacter causing myositis after influenza virus vaccination. PMID- 9225904 TI - Stevens-Johnson syndrome and respiratory failure in a 9-year-old boy. AB - The etiology of respiratory failure associated with Stevens-Johnson syndrome may be multifactorial, including upper airway involvement, pneumothorax/pneumomediastinum, and direct involvement of the respiratory mucosa. Respiratory failure from direct involvement of the respiratory mucosa is relatively uncommon. We describe a 9-year-old boy who had respiratory failure associated with Mycoplasma pneumoniae-induced Stevens-Johnson syndrome. Bronchoscopic examination of the airways revealed sloughed mucosa, ulcerative lesions, and inspissated secretions indicative of lower airway involvement with Stevens-Johnson syndrome. Although the mainstay of therapy is supportive care with controlled ventilation, rigid bronchoscopy with bronchoalveolar lavage to clear the airways of the debris was an invaluable adjunct to this patient's care. PMID- 9225905 TI - Presentation and management of a thyroglossal duct cyst with a papillary carcinoma. AB - Thyroglossal duct carcinoma is rare, and its presentation is similar to that of a thyroglossal duct cyst: a nontender, palpable mass in the midline location. Rapid increase in growth may be a sign of malignancy, but the diagnosis is based on the pathology of the cyst. Initial treatment of thyroglossal duct carcinoma is the same surgical procedure used for removal of a thyroglossal duct cyst. Further surgery depends on the finding of thyroid nodules or positive lymph nodes but is rarely necessary. The recurrence rate after simple excision is low. Postoperative radioiodine ablation or radiation is considered in cases of recurrence or metastasis. In this report, we describe a patient with a new-onset, nontender, neck mass who had a Sistrunk procedure for a presumed thyroglossal duct cyst and was found to have papillary carcinoma. PMID- 9225906 TI - Geriatric medicine as a challenge to internists. PMID- 9225907 TI - Implantation of colon cancer at trocar sites is increased by low pressure pneumoperitoneum. AB - BACKGROUND: The purpose of this study was to determine the effect of pneumoperitoneum on the implantation of tumor at trocar sites. METHODS: GW-39 human colon cancer cell suspension (0.5 ml of 2.5% v/v) was injected into the peritoneal cavity of golden Syrian hamsters through a 1 cm midline incision. Four 5 mm trocars were inserted through the anterior abdominal wall, and the midline incision was then closed. The animals were randomized to receive pneumoperitoneum (n = 62) or no pneumoperitoneum (n = 60) for 10 minutes. Tumor implantations at trocar sites and midline wound incisions were documented grossly and histologically 8 weeks later. RESULTS: Tumor was identified in 86% (49 of 57) of control animals and 95% (52 of 55) of the experimental group (p = 0.20). Implants increased with pneumoperitoneum at the midline incision from 44% to 71% (p < 0.01) and at trocar sites from 41% to 64% (p < 0.00001). CONCLUSIONS: Pneumoperitoneum significantly increased tumor implantation at trocar sites and midline incisions. PMID- 9225909 TI - Surgical treatment for primary esophageal cancer developing after pharyngolaryngectomy for head and neck cancer. AB - BACKGROUND: Primary esophageal cancer developing after hypopharyngeal or laryngeal cancer is increasing in frequency, but operative treatment of such cases is not well established. The proximity of both cancers could produce interactive influence on surgical procedures and risk factors. There have been few reports focusing on surgical strategy for such cases. METHODS: We retrospectively investigated the most recent series of 18 patients who had previously undergone pharyngolaryngectomy for cancer and subsequently underwent esophagectomy for second primary cancer of the esophagus. At esophagectomy special care was taken to preserve the tracheal vascularity and to select adequate procedures for lymph node dissection and bowel reconstruction. RESULTS: Curative resection as confirmed macroscopically was achieved in all cases except one. Lymph node involvement was found in half of the patients. No major postoperative complication was observed except for partial necrosis of the trachea in two cases that were conservatively treated. No patient died during hospitalization. The 3-year survival rate was 64% overall and 55% in cases with lymph node involvement. CONCLUSIONS: Although increased operative risk was expected in esophagectomy after pharyngolaryngectomy, our operative results were acceptable. Most risk factors were controllable by selecting appropriate operation procedures. In patients with technically resectable cancer, esophagectomy with systematic lymph node dissection is recommended as in ordinary esophageal cancer. PMID- 9225908 TI - Thoracoscopic esophagectomy for esophageal cancer. AB - BACKGROUND: Minimal access surgery is an alternative to open surgery in esophageal surgery. Its role in cancer resection is controversial. METHODS: Thoracoscopic esophageal resection was attempted in 22 patients who had increased operative risk. Postoperative outcomes of these patients were compared with the outcomes of 63 patients who underwent open thoracotomy resection during the same period. RESULTS: Thoracoscopy was completed in 18 patients. Conversion to thoracotomy was necessary because of locally advanced tumor in three patients, and a bypass procedure was performed in another patient because of poor ventilation during thoracoscopy and the finding of metastatic disease. The median thoracoscopy time was 110 minutes (range, 55 to 165 minutes). The total operating times were 240 minutes (range, 165 to 360 minutes) and 250 minutes (range, 190 to 420 minutes) for thoracoscopy and thoracotomy, respectively, p = 0.5. Blood loss was significantly less than that of open resection; medians were 450 ml (range, 200 to 800 ml) and 700 ml (range, 300 to 2500 ml) for thoracoscopy and thoracotomy, respectively, p < 0.01. The median number of lymph nodes removed at thoracoscopy was 7 (range, 2 to 13) compared with 13 (range, 5 to 34) in the thoracotomy group. Bronchopneumonia affected 17% of both groups of patients. Only one patient who was converted to open thoracotomy died. Port site recurrence developed in one patient. Overall survival rates were not significantly different. CONCLUSIONS: Thoracoscopic esophageal resection was a feasible option. Clear advantages over open thoracotomy were not demonstrated, although patients who were selected for thoracoscopy had worse performance status. This technique deserves further investigation in dedicated centers. PMID- 9225910 TI - Splenectomy in the accelerated or blastic phase of chronic myelogenous leukemia: a single-institution, 25-year experience. AB - BACKGROUND: Patients in the accelerated or blastic phases of chronic myelogenous leukemia (CML) often have painful splenomegaly and secondary thrombocytopenia. We tested the hypothesis that splenectomy can be performed with minimal complications in advanced CML, thereby alleviating pain, reversing thrombocytopenia, and minimizing transfusion requirements. METHODS: We reviewed the records of 53 patients in the accelerated or blastic phases of CML who underwent splenectomy between 1970 and 1995 at the U. T. M. D. Anderson Cancer Center. RESULTS: Twenty-eight patients were in accelerated phase and 25 in blastic phase at the time of splenectomy. The most common indications for splenectomy were symptomatic splenomegaly (median splenic weight, 1000 gm; range, 120 to 6700 gm) or thrombocytopenia (platelet count less than 100,000/microliter) or both. There was 1 death within 30 days of splenectomy. The preoperative platelet count increased 3.72-fold +/- 0.53-fold (mean +/- SEM) by postoperative day 7 (p < 0.001; paired t test). Patients with transfusion-dependent thrombocytopenia had significantly fewer platelet and red blood cell transfusions in the 6 months after splenectomy than in the 6 months before splenectomy (p = 0.016; sign test). CONCLUSIONS: Splenectomy can be performed with minimal morbidity and mortality in advanced CML, thereby relieving symptomatic splenomegaly, reversing thrombocytopenia, and minimizing transfusion requirements. PMID- 9225911 TI - Popliteal artery entrapment syndrome: the role of early diagnosis and treatment. AB - BACKGROUND: The purpose of this study was to evaluate whether certain factors could influence arterial impairment at presentation for treatment of popliteal artery entrapment syndrome (PAES) and whether its early diagnosis could optimize long-term results. METHODS: Between 1979 and 1995, 30 patients were treated for PAES at our institution. Patients were characterized by age, risk factors, associated diseases, preoperative symptoms, affected side, dominant limb, duration of symptoms, musculotendinous structure causing the compression, arteriographic findings, arterial status at presentation, type of operation, postoperative complications, and long-term follow-up. RESULTS: Twenty-nine (65%) limbs underwent musculotendinous section (MTS), 15 (33%) limbs underwent vascular reconstruction, and 1 (2%) was surgically explored. Patients submitted to MTS were younger (mean, 31 +/- 3 years) than patients who underwent vascular reconstruction (mean, 41 +/- 4 years; p < 0.05). MTS limbs had a greater number of minor symptoms compared with those that underwent vascular reconstruction (62% versus 20%; p < 0.02). Arteriogram showed that MTS limbs had a greater number of normal findings at rest when compared with limbs that underwent conventional reconstruction (85% versus 0%; p < 0.001). No specific factors influenced the arterial status at presentation. During follow-up, treadmill examination revealed that MTS limbs had a better response (96%) than limbs that had undergone vascular procedures (67%; p < 0.02). MTS limbs had a better long-term patency rate (mean, 87 +/- 7 months) compared with limbs that were submitted to vascular reconstruction (mean, 107 +/- 8 months) (95% versus 65%; p < 0.02). CONCLUSIONS: Because PAES is a progressive disease that can create serious vascular obstructive disease and no specific factors seem to influence the degree of vascular impairment, the detection and treatment of PAES at an early stage permit better long-term results. PMID- 9225912 TI - Hemodynamic effects of isovolemic hemodilution during descending thoracic aortic cross clamping and lower torso reperfusion. AB - BACKGROUND: Isovolemic hemodilution has been suggested for blood conservation and to improve hemodynamic tolerance to abdominal aortic cross clamping. However, the hemodynamic effects of hemodilution during descending thoracic aortic cross clamping (DAC) have not been established. We evaluated them in anesthetized swine. METHODS: Hemodilution (n = 7) was produced by the isovolemic exchange of blood for 6% hetastarch to a target hematocrit of 20%. Hematocrit in control pigs (n = 7) remained at 30%. DAC was performed at the T9 level for 45 minutes. During a 60-minute reperfusion period, control pigs were infused with lactated Ringer's solution; shed blood was returned to hemodilution pigs, followed by lactated Ringer's. If hypotension occurred despite left atrial pressure of 10 mm Hg or greater, boluses of phenylephrine were given to keep mean arterial pressure above 60 mm Hg. RESULTS: Hemodilution caused a marked reduction in hematocrit and in global oxygen delivery (DO2). DAC produced a significant increase in proximal arterial pressure, cardiac index, and DO2 and oxygen consumption (VO2) was markedly reduced in both groups. A significant increase in systemic vascular resistance during DAC occurred only in control pigs. After reperfusion, vascular resistance was significantly lower than baseline in hemodilution pigs, requiring a sixfold greater dose of phenylephrine to avoid hypotension. A lower global DO2 and supply-limited VO2 were also observed in hemodilution pigs. CONCLUSIONS: Isovolemic hemodilution maintains hemodynamic stability during DAC. During lower torso reperfusion, however, hemodilution caused hemodynamic instability, decreased global DO2, and limited VO2, which may offset its potential benefits. PMID- 9225913 TI - Elastin degradation products induce adventitial angiogenesis in the Anidjar/Dobrin rat aneurysm model. AB - BACKGROUND: Infusion of the abdominal aorta with pancreatic elastase induces aneurysms in a rat model (Anidjar/Dobrin). Because elastolysis liberates elastin degradation products (EDPs), the present experiment was carried out to test the hypothesis that an EDP alone could induce features of aneurysm disease. METHODS: The EDP val/gly/val/ala/pro/gly (VGVAPG), elastase, or saline solution was infused into infrarenal aorta (n = 4/group). After 1 week aortic diameters were measured, and the tissues were prepared for histologic examination. Adventitial capillaries (vessels per high-power field) were counted over a standardized preparation of aorta. Wall thickness was measured by means of computer-aided planimetry. RESULTS: There was an increase of greater than 100-fold in mean vessels per high-power field in aortas receiving VGVAPG or elastase versus saline controls (4.10 +/- 0.68 SEM or 4.48 +/- 0.49 SEM versus 0.03 +/- 0.03 SEM, respectively, p < 0.05). The VGVAPG-perfused group had a 26% +/- 4% SEM increase in diameter from baseline that was statistically significant (p < 0.01), but the aortas did not reach aneurysmal dimensions. CONCLUSIONS: Although no aneurysms occurred at 1 week after the infusion of EDP, the results demonstrate that the EDP VGVAPG can induce a characteristic feature of aneurysm disease. The model permits study of the earliest stages of experimental aneurysm formation and raises interesting questions regarding the role of the vasa vasorum in this pathologic process. PMID- 9225914 TI - Protective effect of monoclonal antibodies to adhesion molecules on rat liver ischemia-reperfusion injury. AB - BACKGROUND: The source of reactive oxygen species in the liver remains to be elucidated. The present study was undertaken to determine whether polymorphonuclear neutrophils (PMNs) can contribute to hepatic ischemia reperfusion injury, and pretreatment with monoclonal antibodies (mAbs) to intercellular adhesion molecule-1 (ICAM-1), lymphocyte function associated antigen-1 (LFA-1), and CD 18 could improve energy metabolism and prolong the viability of the organ. METHODS: Male Wistar rats were used. Rat liver ischemia was induced by clamping blood vessels supplying median and left lateral hepatic lobes. Monoclonal antibodies to ICAM-1, LFA-1, or CD18 were injected intravenously 5 minutes before inducing ischemia. To determine the effect of mAbs on the survival rate, total hepatic ischemia was induced by clamping the hepatic artery, portal vein, and bile duct after making a portafemoral shunt. RESULTS: Although ischemia of the liver for 90 minutes did not permit survival of the animals, pretreatment with mAbs to ICAM-1 plus LFA-1 increased the survival rate to 57%. Pretreatment with mAb to ICAM-1 failed to increase the survival rate. The number of PMNs in the liver increased continually up to 24 hours after reperfusion after 90 minutes of ischemia, and the expression of ICAM-1 was enhanced 4 hours after reperfusion. This is accompanied by a low recovery of hepatic adenosine triphosphate and, on the contrary, a marked increase in lipid peroxide in the reperfused liver. Pretreatment with mAbs suppressed the infiltration of PMNs and the elevation of lipid peroxide and enhanced the recovery of hepatic adenosine triphosphate 6, 12, or 24 hours after reperfusion. Pretreatment with mAbs also prevented the rise in serum alanine aminotransferase level after reperfusion. CONCLUSIONS: These results suggest that PMNs contribute to ischemia-reperfusion injury in the liver 4 hours and more after reperfusion, and pretreatment with mAbs to adhesion molecules is useful for the prevention of ischemic liver cell injury. PMID- 9225915 TI - Pain as a predictor of outcome in patients with operable pancreatic carcinoma. AB - BACKGROUND: The purpose of our study was to evaluate the relationship between pain and resectability and survival in patients with operable pancreatic carcinoma. METHODS: Pain, pain intensity, and pain location were prospectively assessed in newly diagnosed patients with operable adenocarcinoma of the pancreas. Patients were evaluated before their first operation at a large tertiary care cancer center. Pain factors were then correlated with outcomes of surgery, including resectability and survival. RESULTS: Seventy-seven patients with operable pancreatic carcinoma were evaluated before operation. With the Memorial Pain Assessment Card and a demographic questionnaire, an analysis of analgesic use and pain prevalence and intensity were quantitated. Twenty-two (29%) of 77 patients reported no pain. Fifty-five had mild to severe pain. Moderately severe or greater pain (Memorial Pain Assessment Card Tursky scores of 5 or greater or visual analogue self-assessment pain intensity scores greater than 30) was found in 20 patients. Twenty-six (34%) patients had resectable disease. Of the 51 patients who did not have resections, 35 had metastatic disease. Locoregional unresectable disease without metastases was found in 16 patients. Resectability was correlated with the presence of pain (p = 0.04). The median survival for all patients was 6.7 months. Not surprisingly, patients undergoing resection had a significantly better median survival than did those whose disease was unresectable (5.5 versus 15.1 months). Pain before operation significantly predicted survival (median survival for those with pain, 5.7 months; for those without pain, 15 months; p = 0.003). Even among patients who underwent resection, the presence of pain was associated with a worse survival (21.9 months versus 9.2 months; p = 0.045). In a multivariate analysis the two significant variables were inability to undergo resection and presence of any pain. CONCLUSIONS: The presence of pain in newly diagnosed patients with potentially operable pancreatic cancer is an ominous predictor of resectability and of survival. Even if the patient can undergo resection, the presence of preoperative pain is associated with a poor prognosis. Patients with operable pancreatic cancer who present with pain, even those whose evaluation shows a likelihood of resectability, are at high risk for recurrence with an impaired survival compared with those patients without pain. PMID- 9225916 TI - Anti-interleukin-8 monoclonal antibody reduces free radical production and improves hemodynamics and survival rate in endotoxic shock in rabbits. AB - BACKGROUND: Although high levels of interleukin-8 (IL-8) have been found in patients with sepsis and a monoclonal antibody (MoAb) against IL-8 has been successfully used in some animal models of inflammation, no specific therapeutic agent against IL-8 has been tested for the treatment of sepsis. We studied the effects of a MoAb against IL-8 in the treatment of endotoxic shock with a prospective randomized rabbit endotoxic shock model. METHODS: Twenty New Zealand white rabbits were anesthetized and divided into four groups: normal, anti-IL-8, control-Ab, and lipopolysaccharide (LPS). Anti-IL-8 and control-Ab groups received a MoAb (immunoglobulin G, 3 mg/kg) 5 minutes before the LPS injection. All groups, except the normal group, received a continuous 20-minute infusion of LPS (500 micrograms/kg). The normal group received NaCl (0.9%) rather than LPS. RESULTS: The 7-day survival rates were 100% for normal group, 80% for anti-IL-8 group, 40% for control-Ab group, and 0% for LPS group. Compared with the LPS group, anti-IL-8 rabbits had a smaller decrease in mean arterial blood pressure (p < 0.05) and increased urinary volume (p < 0.05). Anti-IL-8 rabbits had lower plasmatic levels of IL-1 beta, less free radical production (p < 0.05), and a higher survival rate (p < 0.01). CONCLUSIONS: IL-8 plays a significant role in endotoxic shock, and IL-8 blockage results in attenuation of the hypotensive and tachypneic effects of LPS, reduced free radical production, and an increased survival rate after lethal endotoxic shock. PMID- 9225917 TI - Octylcyanoacrylate tissue adhesive versus suture wound repair in a contaminated wound model. AB - BACKGROUND: Octylcyanoacrylate tissue adhesive is a topical wound closure that precludes the need for foreign bodies (sutures) to close wounds. It also has an in vitro antimicrobial effect when standard disc sensitivity tests are used. METHODS: To determine whether contaminated wounds closed with octylcyanoacrylate tissue adhesive will have a lower infection rate compared with wounds closed with 5-0 monofilament sutures, we designed a randomized, blinded, experimental animal study. Two incisions were made on 20 albino guinea pigs. The wounds were contaminated with 10(5) Staphylococcus aureus ATCC 12600 and randomly assigned to be closed with either topical octylcyanoacrylate tissue adhesive or percutaneous 5-0 polypropylene suture. Five days later the adhesive and sutures were removed, and a section of the wound was given to a histopathologist blinded to the type of wound closure. The wound was determined to be infected if inflammatory cells with intracellular cocci were seen. The rest of the wound was opened and examined for clinical evidence of infection. Quantitative bacteriologic analysis was performed. RESULTS: Five wounds in the tissue adhesive group were sterile on day 5, whereas all sutured wounds had positive cultures (25% versus 0%, p < 0.05). Fewer wounds in the tissue adhesive group were determined to be infected by histologic and clinical criteria (0% versus 55%, p < 0.001, and 20% versus 65%, p < 0.01, respectively). Agreement on the determination of infection by histologic and clinical criteria yielded a kappa coefficient of 0.46 (95% confidence interval [GI], 0.19 to 0.73). An infection criterion of 10(5) colony-forming units/gm of tissue correlated poorly with clinical and histologic infection rates (0.19 [95% CI, -0.06 to 0.44] and 0.13 [95% CI, -0.05 to 0.31], respectively). CONCLUSIONS: Contaminated wounds closed with sutures had higher infection rates compared with those reported with topical tissue adhesive. The amount of colonization may not be an accurate method to determine infection. PMID- 9225918 TI - The immunogenicity of the extracellular matrix in arterial xenografts. AB - BACKGROUND: Determinants of xenograft immunogenicity are poorly characterized. We showed previously that decellularized arterial xenografts (DAXs) dilate, whereas decellularized arterial isografts (DAIs) and allografts do not, suggesting an interspecies, rather than an intraspecies, immunogenicity of the arterial extracellular matrix leading to chronic rejection. Now we have investigated the immunogenicity of the arterial extracellular matrix in xenografts and its impact on chronic injury (elastin lysis) and remodeling (graft dilation). METHODS: Diameter and elastin content were measured in DAIs and DAXs from hamster to rat (concordant combination) and guinea pig to rat (discordant combinations) at 8 weeks. We also characterized the immune effectors infiltrating DAIs and DAXs by immunohistochemistry after 6 hours to 4 weeks of implantation. Results were compared with nondecellularized isografts and xenografts. Last, the impact of the donor-recipient phylogenetic distance on monocyte-macrophage penetration into the media was assessed in three xenograft combinations. RESULTS: DAXs from guinea pig, but not from hamster, were aneurysmal at 8 weeks. Elastin lysis paralleled graft dilation. DAXs, but not DAIs, were infiltrated by monocytes, macrophages, T lymphocytes, and immunoglobulins. The donor-recipient combination did not affect the phenotype of the inflammatory infiltrate in DAXs, but it modified the kinetics of monocyte-macrophage penetration into the media. The absence of decellularization changed the inflammatory infiltrate phenotype (absence of macrophages) but had little impact on DAX injury and remodeling. CONCLUSIONS: DAX immunogenicity accounts for most of chronic arterial xenograft injury, which is modulated by the donor-recipient combination. The immunogenicity of arterial xenografts, unlike allografts, is supported by the extracellular matrix in addition to the cells and could influence the long-term fate of xenografts. PMID- 9225919 TI - A rat model of pancreatic ductal adenocarcinoma: targeting chemical carcinogens. AB - BACKGROUND: Current experimental models of pancreatic cancer either fail to reproduce the ductal phenotype or cause simultaneous cancers in other organs also. To develop an animal of pancreatic cancer that accurately mimics the human condition, we restricted carcinogenic exposure to the pancreas and specifically targeted ductal epithelial cells. Three different carcinogens were either implanted directly into the pancreas or infused into the pancreatic duct, with or without near-total pancreatectomy (as a means of inducing pancreatic ductal cell proliferation). METHODS: Groups of male Sprague-Dawley rats were exposed to varying doses of dimethylbenzanthracine (DMBA), methynitronitrosoguanidine, or ethylnitronitrosoguanidine either through direct implantation into the pancreas or infusion into the pancreatic duct. Near-total pancreatectomy was added in all groups except two DMBA implantation groups. Surviving rats were killed at 3, 6, 9, or 12 months, and the pancreata were evaluated histologically. RESULTS: All three carcinogens caused pancreatic inflammation, ductal hyperplasia, atypia, and dysplasia beginning by 3 months and becoming more prominent at later time points. Only DMBA caused frequent invasive pancreatic ductal adenocarcinoma, which was first evident by 6 months. The prevalence of pancreatic cancer among DMBA-treated rats evaluated after 10 months was 39% (19 of 49). The addition of pancreatic resection did not enhance pancreatic cancer development. CONCLUSIONS: Of the strategies tested, only direct implantation of DMBA into the rat pancreas frequently produces pancreatic cancer histologically similar to human ductal adenocarcinoma. The development of hyperplastic, atypical, and dysplastic changes preceding and accompanying carcinomas suggests that these lesions are preneoplastic. This model recapitulates the progression from normal to neoplastic epithelium and is likely to be useful for the study of morphologic and molecular mechanisms underlying the early stages of pancreatic carcinogenesis and for the investigation of novel diagnostic and therapeutic techniques. PMID- 9225920 TI - Adenovirus-mediated transfer of tissue-type plasminogen activator gene to human endothelial cells. AB - BACKGROUND: Seeding of vascular grafts with genetically engineered human endothelial cells (hECs) secreting antithrombogenic or fibrinolytic agents has considerable clinical potential. METHODS: An adenoviral vector was used to transfer the human tissue-type plasminogen activator (htPA) gene to hECs, and the ability of the transduced hECs to secrete htPA was examined. Cultured hECs on plates were incubated with various concentrations of recombinant adenoviruses containing the htPA or LacZ gene for various times to determine the optimal transfer conditions. Transduced hECs were seeded onto fibronectin-coated expanded polytetrafluoroethylene grafts (4 mm in diameter), some of which were exposed to pulsatile flow in vitro. RESULTS: Effective transduction of the htPA gene into hECs (htPAhECs) was achieved with viral soup at a multiplicity of infection of 30 after incubation for 1 day, which yielded 4.8 +/- 0.20 x 10(3) ng/10(6) cells/6 hr htPA antigen on plates (n = 3), 2.2 +/- 2.0 x 10(3) ng/10(6) cells/6 hr on grafts (n = 6), and 6.8 +/- 1.7 x 10(2) ng/10(6) cells/6 hr on perfused grafts (n = 6). The retention of htPAhECs by perfused grafts was 84.0% +/- 3.0%, comparable with the noninfected (82.1% +/- 8.0%) and mock-infected (94.2% +/- 0.4%) hEC values. CONCLUSIONS: By adenoviral vector-mediated gene transfer, 10(2-3)-fold enhancement of htPA secretion was demonstrated, which did not affect cell retention by grafts. PMID- 9225921 TI - Sun Yat-sen: surgeon and revolutionary. PMID- 9225922 TI - Spindle cell stromal tumor of the pancreas: treatment by pancreatoduodenectomy. PMID- 9225923 TI - Aortic aneurysm rupture after extracorporeal shock wave lithotripsy. PMID- 9225924 TI - Increased risk for inguinal hernia in patients with Ehlers-Danlos syndrome. PMID- 9225925 TI - False-positive parathyroid scan leading to sternotomy: incidental detection of a thymoma by C-11 methionine positron emission tomography. PMID- 9225926 TI - Surgical management of hyperparathyroidism in view of a reliable parathyroid adenoma localization test. PMID- 9225927 TI - Is laparoscopic cholecystectomy indicated for early gallbladder cancer? PMID- 9225928 TI - Cellular gene therapy: a credible odyssey. PMID- 9225929 TI - Generation of cytokines in red cell concentrates during storage is prevented by prestorage white cell reduction. AB - BACKGROUND: The effect of prestorage white cell (WBC) filtration on the cytokine content in red cells (RBCs) has not been clarified. STUDY DESIGN AND METHODS: Six units of buffy coat-poor RBC concentrates were prepared. Each unit was divided into three parts: one was used as a control, the second was made WBC-rich by the addition of WBCs from the buffy coat, and the third was made WBC-poor by filtration. All units were stored at 4 degrees C for 6 weeks. Immediately after preparation and every second week subsequently, samples for analyses of interleukin (IL) 1 beta (IL-1), IL-2, IL-6, IL-8, and tumor necrosis factor alpha (TNF alpha) were obtained. After 13 weeks, the procedure was repeated on the same donors. RESULTS: IL-2 was not detected. The amounts of IL-1, IL-8, and TNF alpha increased during the storage period. With the exception of IL-8, only low concentrations were found. Filtered units had lower concentrations of IL-1, IL-6, IL-8, and TNF alpha after 2 weeks of storage than did the control and WBC-rich units. The amounts of cytokines in filtered units did not increase during the study period. CONCLUSION: Prestorage filtration seems to diminish the amount of IL-1, IL-6, IL-8, and TNF alpha RBCs during storage. The possible clinical implications of this should be elucidated. PMID- 9225930 TI - Retroviral transduction of peripheral blood leukocytes in a hollow-fiber bioreactor. AB - BACKGROUND: Peripheral blood white cells (leukocytes) (PBLs) have been used as effective targets for genetic manipulation by transduction with retroviruses in open systems. A semi-closed hollow-fiber bioreactor was tested for culturing and transducing lymphocytes. STUDY DESIGN AND METHODS: PBLs were isolated from five healthy donors, and 5 x 10(7) cells were cultured in hollow-fiber bioreactors for 4 days after stimulation with anti-CD3 in medium containing 200 units per mL of recombinant interleukin 2. Transduction with retroviral vector containing the gene for iduronate-2-sulfatase and G 418 resistance, L2SN, was performed daily on Days 4, 5, 6, and 7, and the cells were expanded for an additional 3 days. RESULTS: PBLs from three donors were harvested from the bioreactor after transduction and expansion, and 4.5 x 10(9), 7.0 x 10(9), and 2.9 x 10(9) cells were recovered, representing 90-, 136-, and 58-fold expansions. The transduction frequency of L2SN was 10, 5, and 1 percent, respectively. For additional expansion of PBLs, in two cases the bioreactor was reinoculated with 5 x 10(7) cells, which were expanded again for 16 and 8 days, respectively, yielding 1.4 x 10(9) and 3.1 x 10(9) cells, which reflected an additional 28- and 62-fold expansion of cells. PBLs from two other donors were transduced and expanded in the bioreactor, and then 0.8 mg per mL of G 418 was added to the medium in an attempt to enrich the transduced population. Although 2.5 and 10 percent of the cells were transduced, cell death and absence of expansion in the presence of G 418 resulted in final cell lots with viabilities of only 4 and 8 percent. In all cases, the harvested cells tested negative in bacterial and fungal cultures. CONCLUSION: Hollow-fiber bioreactors are an efficient and effective system for the retroviral transduction and expansion of PBLs for clinical gene therapy. PMID- 9225931 TI - Inhibitory effect of 0 degree C storage on the proliferation of Yersinia enterocolitica in donated blood. AB - BACKGROUND: Yersinia enterocolitica is frequently identified in cases of bacterial sepsis due to red cell transfusion. One of the features that makes Y. enterocolitica particularly dangerous is that, unlike most other bacterial contaminants of blood components, this organism can actively multiply in currently recommended refrigerator temperatures (1-6 degrees C). The effect of a colder than normal storage temperature on Y. enterocolitica growth was investigated to determine whether bacteria growth could be reduced or inhibited at 0 degree C. STUDY DESIGN AND METHODS: Twenty-four units of freshly collected donated blood were obtained. Three sets of 7 units each were inoculated with Y. enterocolitica O:3, Y. enterocolitica O:20, and Y. enterocolitica O:5, 27, respectively. The remaining 3 units served as uninoculated controls. Each of the 24 bags was split into two equal aliquots, with one aliquot stored at 4 degrees C and the other at 0 degree C. Bacteria growth was measured twice weekly for 6 weeks. Endotoxin and hemoglobin levels were also measured at selected intervals. RESULTS: Bacteria growth was detected earlier and in higher concentrations in the aliquots stored at 4 degrees C. Twenty-two of the 42 inoculated aliquots had measureable bacteria growth. Thirteen aliquots had been maintained at 4 degrees C, and nine had been stored at 0 degree C. Sixteen of these 22 aliquots were matched pairs. Exponential growth was detected after 14 to 32 days in the 4 degrees C aliquots and after 28 to 39 days in the 0 degree C aliquots. Final bacteria counts were much higher in the 4 degrees C aliquots (10(5)-10(14) colony forming units/mL) than in the 0 degree C aliquots (10(1)-10(4) colony-forming units/mL) on Day 42. Endotoxin was present in all 13 of the 4 degrees C aliquots with actively growing Y. enterocolitica. CONCLUSION: Storage of red cells at 0 degree C markedly prolongs the time required for Y. enterocolitica to achieve exponential grwoth and results in lower concentrations of bacteria. PMID- 9225932 TI - Increased platelet aggregation due to chilling to 20 degrees C is not related to increased sensitivity to agonists. AB - BACKGROUND: Previous studies suggested that chilled platelets have a greater sensitivity to agonists than do platelets aggregated at 37 degrees C. STUDY DESIGN AND METHODS: Washed platelets were aggregated at 20 or 37 degrees C with ADP (0-20 microM), arachidonic acid (0-200 microM), or the thromboxane mimetic U46619 (0-9 nM). RESULTS: Chilling caused a significant (p < or = 0.05) increase in spontaneous platelet aggregation (> 27% at 20 degrees C vs < 5% at 37 degrees C) and spontaneous dense granule release (> 0.01 nM of ATP at 20 degrees C vs. 0 nM of ATP at 37 degrees C), ADP and U46619 caused a significantly greater aggregation response and dense granule release at 20 degrees C, although there was no change in agonist sensitivity of platelets (effective dose of agonist necessary to induce 50% aggregation [ED50]: 1.00 +/- 0.35 microM ADP at 20 degrees C and 1.63 +/- 0.47 microM ADP at 37 degrees C; 8.26 +/- 3.65 pM U46619 at 20 degrees C and 18.97 +/- 4.82 pM U46619 at 37 degrees C). Platelets aggregated with arachidonic acid showed a significant decrease in aggregation and agonist sensitivity at 20 degrees C (ED50 118.7 +/- 44.4 microM) from those at 37 degrees C (ED50 25.6 +/- 7.2 microM), possibly as a result of the reduced enzymatic activity of cyclooxygenase and thromboxane synthase at the lower temperature. CONCLUSION: The data suggested that washed platelets chilled to 20 degrees C and aggregated are not more sensitive to agonists than are 37 degrees C controls, but rather that chilled platelets undergo greater spontaneous aggregation. PMID- 9225933 TI - The presurgical management with erythrocytapheresis of a patient with a high oxygen-affinity, unstable Hb variant (Hb Bryn Mawr). AB - BACKGROUND: Hemoglobin (Hb) Bryn Mawr is an unstable Hb variant resulting in congenital hemolytic anemia. This variant Hb also has an increased affinity for oxygen. The perioperative transfusion management of this disorder is described, and the first genomic analysis of this Hb variant is given. CASE REPORT: An 11 year-old boy, heterozygous for Hb Bryn Mawr, was referred for cholecystectomy. Sequence analysis of genomic DNA confirmed that the patients was heterozygous for a T-->C transition in the codon for amino acid 85, causing a substitution of serine for phenylalanine in the beta-globin chain. On the basis of whole-blood O2 dissociation studies, projected tissue O2 delivery would have been suboptimal during general anesthesia; therefore, a partial red cell exchange transfusion was performed to lower variant Hb and prevent tissue hypoxia during surgery. The red cell mass to be exchanged (50%) was determined from the calculated increase in O2 delivery capacity required to maintain an O2 extraction of 4 to 5 mL of O2 per dL of whole blood. The p50 of whole blood from the patients immediately after the exchange transfusion was 16.0 torr. At the time of surgery, the p50 was normal (25.9 torr). The patient's whole blood 2,3 DPG levels were 4.70 mmol per mL of red cells (before transfusion) (normal range = 4.8 +/- 0.3 mmol/mL red cells), 4.07 mmol per mL of red cells (immediately after transfusion), and 4.55 mmol per mL of red cells (48 hours after transfusion). CONCLUSION: This patient with Hb Bryn Mawr was prepared for surgery with a partial exchange transfusion to prevent tissue hypoxia during anesthesia. Decreased 2,3 DPG levels immediately after transfusion resulted in increased O2 affinity of whole blood; however, 48 hours after exchange transfusion, a normal p50 (due to both removal of variant Hb and regeneration of 2,3, DPG) was observed. Partial exchange transfusion is useful in the preoperative management of patients with Hb variants characterized by increased O2 affinity. PMID- 9225934 TI - Recombinant human erythropoietin as adjuvant treatment for autologous blood donation in elective surgery with large blood needs (> or = 5 units): a randomized study. AB - BACKGROUND: Autologous blood transfusion presents no infectious or immunologic side effects. The aim of this randomized study was to determine the impact of recombinant human erythropoietin (rHuEPO) on the donation of 5 units of autologous blood by nonanemic patients who were candidates for elective surgery with transfusion requirements of > or = 5 units. STUDY DESIGN AND METHODS: Starting on Day -35, 420 mL of blood was taken weekly. All patients received 200 mg of iron saccharose complex intravenously at each visit and six subcutaneous injections of rHuEPO (141 U/kg) or placebo between Days -21 and -7. RESULTS: Of 50 patients, 45 completed the study (placebo, 21; rHuEPO, 24). Total red cell production was higher in the rHuEPO group (p = 0.001). Donation of 5 units was possible for 67 percent (placebo group) and 79 percent (rHuEPO group) of patients (p = 0.5). The mean number of blood units donated was 4.6 (placebo group) and 4.7 (rHuEPO group). More patients in the placebo group received allogeneic blood (9/21 [43%] vs. 6/23 [26%]), although the difference did not reach significance (p = 0.34). CONCLUSION: In nonanemic patients donating 5 units of blood, rHuEPO associated with intravenous iron increased total red cell production. However, no difference was found between the rHuEPO and placebo groups with regard to the number of units of autologous blood donated of the number of patients receiving allogeneic blood transfusion. PMID- 9225935 TI - Plateletpheresis in 90- to 110-pound donors using the CS-3000 blood cell separator. AB - BACKGROUND: Increases in the use of single-donor apheresis components have increased the need for platelet donors. In the United States, persons must weigh 110 pounds or more to qualify as blood donors, and the same weight limitation has been placed on apheresis donors. Because automated plateletpheresis with some instruments differs considerably from whole-blood donation with respect to the volume of blood removed from the donor, the feasibility of using persons weighing between 90 and 110 pounds as platelet donors was evaluated by the use of the CS 3000 blood cell separator. STUDY DESIGN AND METHODS: The study was performed using female subjects who met all usual donor requirements except for minimum weight. The standard platelet collection procedure of the instrument was used, except that the blood processing rate was manually selected so as to optimize the blood withdrawal and return rate in individuals. Vital signs were recorded before and after donation as were signs or symptoms of any type of donor reaction. RESULTS: Twenty-six of 28 women completed the donation procedure; in two instances, collection was terminated prematurely because of an inability to maintain adequate venous access. An average of 4.5 x 10(11) platelets were collected during a mean donation time of 110 minutes. All donors tolerated the procedure well, and no serious adverse reactions were seen. Because of the administration of priming solution and anticoagulant during apheresis, there was a net positive fluid balance following the procedure, in spite of the removal of approximately 220 mL of platelet concentrate. CONCLUSION: These preliminary studies suggest that 90- to 110-pound persons may serve as plateletpheresis donors. Additional studies are needed to more fully document the safety and efficacy of this approach. The use of lower-weight donors may significantly increase the number of persons available to provide single-donor platelet components. PMID- 9225936 TI - The association of biologically active lipids with the development of transfusion related acute lung injury: a retrospective study. AB - BACKGROUND: Transfusion-related acute lung injury (TRALI) is clinically similar to the adult respiratory distress syndrome (ARDS) and has been linked to the transfusion of leukocyte antibodies in blood components. Animal model have implicated neutrophil (PMN)-priming agents in ARDS; however, two agents were required. Previous studies showed the generation of PMN-priming agents during blood storage. Thus the association of PMN-priming agents with TRALI was examined. STUDY DESIGN AND METHODS: Ten patients with TRALI and 10 with febrile or urticarial reactions (control group) were evaluated. The presence of PMN priming activity was tested in the patients' pretransfusion and posttransfusion blood samples by incubating PMNs with these samples followed by activation of the respiratory burst. Plasma lipids were separated by normal-phase high-performance liquid chromatography (HPLC), and the priming activity was evaluated. The presence of leukocyte antibodies was determined in the blood donors and patients with TRALI. RESULTS: Significantly more PMN-priming activity was present in the posttransfusion sera (11.4 +/- 1.8 nmol superoxide anion/min, mean +/- SEM; n = 10) and plasma of patients with TRALI than in their pretransfusion sera (6.5 +/- 1.5: n = 10) or in the pretransfusion and posttransfusion sera (5.1 +/- 1.3, n = 10; and 4.5 +/- 1.4, n = 10, respectively) and from the controls (p < 0.05). HPLC separation of lipids demonstrated that three active species were present in the posttransfusion plasma samples of TRALI patients. All the patients with TRALI had underlying clinical factors, such as infection, cytokine administration, recent surgery, or massive transfusion, while only 2 of 10 control patients had these clinical conditions. None of the donors had significant titers of HLA or HLA-DR antibodies; however, 50 percent had weak positivity for granulocyte antibodies. CONCLUSION: TRALI is the result of two clinical events, the first being a predisposing clinical condition and the second being the transfusion of biologically active lipids in stored blood. PMID- 9225937 TI - Specific antibodies to Trypanosoma cruzi among blood donors in Los Angeles, California. AB - BACKGROUND: Trypanosoma cruzi, the cause of Chagas' disease, is often transmitted by transfusion in Latin America. Previous studies showed that at least 1 in 1000 eligible blood donors at the Los Angeles County+University of Southern California (LAC+USC) Medical Center Blood Bank had specific antibodies to T. cruzi. In June 1993, serologic screening of prospective allogeneic donors at epidemiologic risk for T. cruzi infection was begun voluntarily. STUDY DESIGN AND METHODS: The risk of T. cruzi infection in all eligible donors was assessed by questionnaire. At risk donors were screened serologically for antibodies to T. cruzi with an enzyme immunoassay, and confirmatory testing was done with a radioimmunoprecipitation assay. RESULTS: During the 29-month study period 1311 (39.5%) of 3320 donors were judged to be at risk for T. cruzi infection. Seven donors (1/475) were reactive by an enzyme immunoassay, and six of these seven (1/ 553) were positive in a radioimmunoprecipitation assay. All radioimmunoprecipitation assay-positive donors had been born in countries in which Chagas' disease is endemic. One person in this group had received a transfusion in his homeland. CONCLUSION: These results demonstrate that a substantive proportion of eligible blood donors at our institution have antibodies specific for T. cruzi and that a commercially available assay can be used to detect these antibodies. Our data suggest that the risk of transmission of T. cruzi by transfusion could be eliminated by serologic testing limited to persons born in or transfused in countries in which Chagas' disease is endemic. PMID- 9225938 TI - A prospective study of a serum-pooling strategy in screening blood donors for antibody to hepatitis C virus. AB - BACKGROUND: To examine the feasibility and to perform a cost-benefit analysis of a pooling protocol of enzyme immunoassay (EIA) screening for antibody to hepatitis C virus (anti-HCV) under real conditions, a prospective study was carried out using sera from 1875 local blood donors. STUDY DESIGN AND METHODS: In the absence of knowledge of the anti-HCV reactions, the donor's sera were pooled into groups of five consecutive samples for testing by EIA. The dilution and final volume of the serum pool were adjusted to equal those recommended for single-serum EIA by the manufacturer of the test kit. The results obtained were compared with those of single-serum EIA to assess the feasibility of the pooling protocol. By applying probability theory, the percentage of reduction in the number of tests performed (L value) when the serum-pooling strategy was used was calculated for several anti-HCV seroprevalences and for varied sizes of pool. The calculations were performed on a computer using a program compiled by the authors. RESULTS: The results showed that seroprevalence was 2.24 percent (95% CI, 1.57-2.91%); the rate of false negativity was 0 (95% CI, 0-8.4%), the sensitivity of the pooling protocol was 100 percent (95% CI, 91.6-100.0%), the rate of false positivity was 0.8 percent (95% CI, 0-1.8%), and the specificity of the pooling protocol was 99.2 percent (95% CI, 98.2-100.0%). Cost-benefit analysis showed that the pooling protocol could save 69.3 percent of the cost. A table of L values can be used conveniently by serologists to determine the optimum pool size if estimates of seroprevalence are available. CONCLUSION: The pool EIA did not perform worse than individual EIAs, and the pooling strategy was markedly less expansive. The pooling protocol was recommended for screening of anti-HCV-positive subjects from large populations with low seroprevalence. PMID- 9225939 TI - Transfusions of granulocyte-colony-stimulating factor-mobilized granulocyte components to allogeneic transplant recipients: analysis of kinetics and factors determining posttransfusion neutrophil and platelet counts. AB - BACKGROUND: Granulocyte-colony-stimulating factor (G-CSF) is a safe and effective agent for mobilization of neutrophils in normal donors, consistently resulting in cell yields per leukapheresis (LA) procedure that are superior to those with other agents. LA components also contain platelets, whose clinical relevance is unknown. STUDY DESIGN AND METHODS: This study describes the kinetics of and analyzes the factors determining the ANC and platelet count increments seen with each of three transfusions of granulocytes collected from HLA-matched sibling donors receiving G-CSF (n = 10; maximum of 3 LA procedures/donor). The transfusions were given to recipients (n = 10) on alternate days beginning. Day 1 after allogeneic bone marrow transplant (BMT). RESULTS: Significant, sustained increments in the recipient ANCs were observed after the transfusion of G-CSF mobilized LA components. The mean peak posttransfusion increments in the ANCs were 1195, 729, and 631 per microL with transfusion of donor LA components on Days 1, 3, and 5, respectively. The length of time that the mean posttransfusion ANC was at or above the baseline (pretransfusion) value was 25 to 37 hours, depending on the post-BMT day when the component was administered. No consistent relationship was observed between LA component granulocyte dose, baseline recipient ANC, or temperature elevation and post-transfusion ANC increments. Large numbers of platelets (mean, 2.55 x 10(11)) were present in LA components, and this resulted in significant increments from baseline in the mean platelet count 1 hour after LA component transfusions. Between Days 1 and 7, the duration of severe neutropenia was shorter and the percentage of patients requiring nondonor platelet transfusions was less in study patients who received LA component transfusions than in a similar historical control group who did not. CONCLUSION: The transfusion of G-CSF-mobilized, HLA-matched LA components to allogeneic BMT recipients resulted in significant and sustained increments in the ANC and the platelet count. Within the range examined, a relationship between neutrophil dose and an increment in the ANC was not demonstrated. PMID- 9225940 TI - A graded scale for assessment of safety of blood substitutes. PMID- 9225942 TI - Out-of-hospital transfusion. AB - Although issues regarding the provision of transfusions in nonhospital settings are still unresolved, there is now a considerable body of experience with such transfusions and a growing consensus that there are acceptable ways to make the service available to carefully selected patients. Besides the core of published information on the subject, there is ongoing interest and a comparison of experiences occurring at meetings such as those of the AABB and state organizations such as the California Blood Bank Society. These formal and informal exchanges of information by blood centers, hospitals, and outpatient facilities continue to improve the quality achieved by those just embarking on out-of-hospital transfusion programs and those already experienced in this approach. The ultimate beneficiaries of all this shared quality improvement are those patients for whom high-quality out-of-hospital transfusion is not only appropriate but even preferable to inpatient care. PMID- 9225943 TI - Citrate ratio in collection of peripheral blood progenitors. PMID- 9225944 TI - Potential usefulness of protease inhibitor and chloroquine in the treatment of transfusion-associated graft-versus-host disease. PMID- 9225945 TI - Biodistribution of an antibody-enzyme conjugate for antibody-directed enzyme prodrug therapy in nude mice bearing a human colon adenocarcinoma xenograft. AB - The enzyme carboxypeptidase G2 (CPG2) can be targeted to tumors by antibodies and used to activate prodrugs in a treatment called antibody-directed enzyme prodrug therapy (ADEPT). Different doses of CPG2 conjugated to the anti-CEA antibody A5B7 were administered i.v. to nude mice bearing the LS174T human colon adenocarcinoma xenograft, and the biodistribution of conjugate activity 48 and 72 h later was determined using a novel high-performance liquid chromatography (HPLC) method. Conjugate doses of 2,500 and 625 U/kg gave tumor enzyme levels of 0.5-0.6 U/g. Lower doses of 300 and 150 U/kg gave tumor enzyme levels of 0.1-0.3 U/g. Intriguingly, the best tumor:blood ratio of conjugate activity at both 48 and 72 h was achieved after administration of the 625-U/kg dose, not the 2,500-U/kg dose. After 48 h this ratio was 3.8, whereas after 72 h the value was 5.5. This conjugate dose also gave the greatest tumor:tissue ratios in all other tissues examined. After 72 h the tumor:colon ratio was 105, whereas the tumor:kidney ratio was 36. In ADEPT, to obtain maximal tumor damage to LS174T xenografts in nude mice with minimal systemic toxicity using the A5B7-CPG2 conjugate, prodrug should therefore be administered at least 72 h after a conjugate dose of 625 U/kg. PMID- 9225946 TI - Bladder tissue pharmacokinetics of intravesical taxol. AB - Our previous studies have suggested that the ineffectiveness of intravesical mitomycin C or doxorubicin therapy against muscle-invading bladder cancer is in part because of the inability of these drugs to penetrate the urothelium (the urothelial drug concentration is < 5% of the concentration in urine). The goal of the present study was to identify agents that are efficiently absorbed across the urothelium. To evaluate the potential use of taxol in intravesical therapy for bladder cancer, we examined the bladder tissue and systemic plasma pharmacokinetics of intravesical taxol in dogs. Animals (approximately 8 kg body weight) were given an instillation of taxol at 500 micrograms in 20 ml water. At 120 min postinstillation, the bladder was emptied and excised, and about 85% of the dose was recovered in the urine. The taxol concentration in the urothelium was about 50% of the concentration in the urine, the concentrations then declined logarithmically in the underlying capillary-perfused tissues. The average tissue concentration (-2 micrograms/g) was two to three times the reported plasma concentration of 0.75 microgram/ml in patients following intravenous infusion of the > 100-fold higher dose of 250 mg/m2. The steady-state plasma concentration was < 0.02% of the average tissue concentration, and was < 0.05% of the maximally tolerated plasma concentration in patients. The octanol:water partitioning coefficients of taxol, doxorubicin, and mitomycin were > 99, 0.52, and 0.41, which parallels the rank order of the partitioning across urothelium, i.e. taxol (approximately 50%) > > doxorubicin approximately mitomycin C (-3%). In summary, the partitioning of taxol across the urothelium was more favorable than the partitioning of mitomycin C and doxorubicin, and the systemic concentration of taxol resulting from intravesical treatment was insignificant in spite of the extensive absorption into the bladder. We conclude that intravesical delivery of taxol provides a significant bladder tissue targeting advantage, and that taxol represents a viable candidate drug for intravesical bladder cancer therapy. PMID- 9225947 TI - Association of cisplatin nephrotoxicity with patient characteristics and cisplatin administration methods. AB - OBJECTIVE: To assess factors that affect cisplatin nephrotoxicity. METHODS: In 425 patients treated with cisplatin, we assessed the effect of pretreatment factors and treatment conditions on the rise in serum creatinine with the first course of cisplatin, on the maximum rise in serum creatinine over the entire course of the cisplatin therapy, and on residual nephrotoxicity after the last cisplatin treatment ended. (Because of the nature of the relationship between serum creatinine and creatinine clearance, rise in serum creatinine was divided by pretreatment creatinine squared.) Patients were dichotomized into the upper quartile versus the lower three quartiles of degree of nephrotoxicity. Multivariate analyses were based on logistic regression, controlling for cisplatin dose per course. RESULTS: Controlling for cisplatin dose per course, factors most closely associated with nephrotoxicity during the first course of cisplatin were: serum albumin and potassium, body surface area, and administration of cisplatin over 2-5 days per course vs 1 day (negative associations). Controlling for cisplatin dose per course, the single factor most closely associated with maximum life-time cisplatin nephrotoxicity was concurrent use of a vinca alkaloid (negative association). Controlling for cisplatin dose per course, factors most closely associated with residual nephrotoxicity after the end of cisplatin therapy were cumulative dose of cisplatin, concurrent use of metoclopramide (positive associations), uric acid and concurrent use of phenytoin and a vinca alkaloid (negative associations). The association of nephrotoxicity with uric acid and with body surface area was felt to be an artifact resulting from its positive association with pretreatment serum creatinine. Nephrotoxicity during the first course of cisplatin also correlated significantly with autopsy kidney cortex platinum concentrations in 77 evaluable patients. CONCLUSIONS: (1) While several factors correlated with cisplatin nephrotoxicity, most of the observed nephrotoxicity was not explained by the variables identified. (2) While most patients received intravenous hydration, patients receiving high hydration volumes did not have significantly less nephrotoxicity than patients receiving lower hydration volumes: (3) Of the variables identified, serum albumin, metoclopramide and phenytoin may have affected nephrotoxicity by altering cisplatin uptake into or distribution within the kidney. PMID- 9225948 TI - Intratumor distribution of doxorubicin following i.v. administration of drug encapsulated in egg phosphatidylcholine/cholesterol liposomes. AB - PURPOSE: A pharmacological evaluation of an egg phosphatidylcholine/cholesterol (55:45 mole ratio, EPC/Chol) liposome doxorubicin formulation was carried out. The objective was to define liposomal lipid and drug distribution within sites of tumor growth following intravenous (i.v.) administration to female BDF1 mice bearing either Lewis lung carcinoma, B16/BL6 melanoma, or L1210 ascitic tumors. METHODS: Mice were injected i.v. with EPC/Chol liposomal doxorubicin, and plasma and tumor levels of lipid and drug were determined 1, 4 and 24 h late with radiolabeled lipid and fluorimetry or fluorescence microscopy, respectively. In addition, single-cell suspensions of the Lewis lung and B16/BL6 tumors were prepared and the presence of macrophages was determined with an FITC-labeled rat antimouse CD11b (MAC-1) antibody. RESULTS: For mice bearing the Lewis lung solid tumors, there was a time-dependent accumulation of liposomal lipid, with a plateau of approximately 500 micrograms lipid/g tumor at 48 h. In contrast, the apparent plateau (microgram doxorubicin/g tumor) for doxorubicin was achieved at 1 h and remained constant over a 72-h time course. In comparison with free drug administered at the maximum tolerated dose (MTD, 20 mg/kg) doxorubicin levels in tumors were two- to threefold greater when the drug was administered in liposomal form. The increase in drug delivery was comparable for both solid tumors. With animals bearing the L1210 ascitic tumor, drug exposure was as much as ten times greater (in comparison with free drug) when doxorubicin was administered in liposomes. An evaluation of single-cell suspensions prepared from the two solid tumors suggested that more than 98% of the tumor-associated drug and liposomal lipid was not tumor cell-associated. Histological studies with the Lewis lung carcinoma, however, revealed that a proportion of the drug did colocalize with tumor-associated macrophages. Analysis of cells obtained from mice bearing ascitic tumors showed that more than 80% of the cell-associated drug could be removed by procedures designed to remove adherent cells. CONCLUSION: The results summarized here suggest drug concentrations within a solid tumor, such as the Lewis lung carcinoma, are constant over time when the drug is given in a "leaky" EPC/Chol formulation. The results also suggest that liposomal lipid within sites of tumor growth is primarily localized within the interstitial spaces or tumor associated macrophages. PMID- 9225949 TI - Pharmacokinetics of intrapericardial administration of 5-fluorouracil. AB - A 30-year-old patient with metastatic breast adenocarcinoma was diagnosed as having a malignant pericardial effusion. METHODS: The patient was treated with two courses of 200 mg 5-fluorouracil (5-FU) followed by 20 mg cisplatin 5 h later directly infused into the pericardial space through a catheter. The drug levels of the 5-FU were monitored during the second treatment. The half-life of 5-FU in the pericardial space was 168.6 min with a concentration of 0.113 mg/ml still detected at 5 h. The area under the curve (AUC) was estimated to be 4.739 mg h/ml. The plasma concentrations of 5-FU ranged from 0.022 to 0.04 mg/ml throughout the infusion. RESULTS: There was no significant change in the patient's blood counts or chemistry profile. She did not experience any side effects during the treatment. A pericardial window was performed 2 days later when balloon pericardiectomy was unsuccessful. The patient eventually succumbed to her disease 4 months later, but without evidence of pericardial effusion. CONCLUSIONS: We conclude that pericardial infusion of 5-FU allowed a high concentration of 5-FU to be achieved within the pericardial sac with a greatly increased half-life over that of systemic 5-FU treatment (168 min vs 6-20 min), and with little systemic toxicity. PMID- 9225950 TI - Combined-modality treatment of inflammatory breast carcinoma: twenty years of experience at M. D. Anderson Cancer Center. AB - PURPOSE: To review the 20 years of experience at M. D. Anderson Cancer Center with a combined-modality approach against inflammatory breast carcinoma. PATIENTS AND METHODS: A total of 178 patients with inflammatory breast carcinoma were treated in the past 20 years at M. D. Anderson Cancer Center by a combined modality approach under four different protocols. Each protocol included induction chemotherapy, then local therapy (radiotherapy or mastectomy), then adjuvant chemotherapy, and, if mastectomy was performed, adjuvant radiotherapy. Chemotherapy consisted of 5-fluorouracil, doxorubicin, and cyclophosphamide (FAC) with or without vincristine and prednisone (VP). In protocol D, patients received an alternate adjuvant chemotherapy regimen, methotrexate and vinblastine (MV), if they did not have a complete response (CR) to induction chemotherapy. RESULTS: The median follow-up of live patients in group A was 215 months, in group B 186 months, in group C 116 months, and in group D 45 months. An estimated 28% of patients were currently free of disease beyond 15 years. At the time of analysis, 50 patients were alive without any evidence of disease. A further 12 patients died of intercurrent illness, and 15 patients were followed beyond 10 years without recurrence of disease. Among initial recurrence, 20% of patients had local failure, 39% systemic failure, and 9% CNS recurrence. Initial response to induction chemotherapy was an important prognostic factor. Disease-free survival (DFS) at 15 years was 44% in patients who had a CR to induction chemotherapy, 31% in those who had a partial response (PR), and 7% in those who had less than a PR. There was no improvement in overall survival (OS) or DFS among patients who underwent alternate chemotherapy (MV) compared with those who did not. Using surgery and radiotherapy as opposed to radiotherapy alone as local therapy did not have an impact on the DFS or OS rate. CONCLUSION: These long-term follow-up data show that with a combined-modality approach a significant fraction of patients (28%) remained free of disease beyond 15 years. In contrast, single modality treatments yielded a DFS of less than 5%. Thus, using combined-modality treatment (chemotherapy, then mastectomy, then chemotherapy and radiotherapy) is recommended as a standard of care for inflammatory breast carcinoma. PMID- 9225951 TI - The vascular compartment hampers accurate determination of teniposide penetration into brain tumor tissue. AB - After a pre-operative 1-h i.v. infusion of 150 mg/m2 of teniposide (Vumon; VM26), the drug levels were determined in resected brain tumor specimens from three patients with malignant glioma and from three patients with brain metastases. Tissue dissections were performed within 0-2.5 h after drug administration in three patients and after 24 h in the other three patients. Teniposide was quantified by high-performance liquid chromatography and the levels of albumin in the resected tissue samples were quantified by radial immunodiffusion. In addition, albumin levels were quantified in normal brain tissue, in malignant glioma and in metastatic brain tumor tissue obtained post mortem from deceased patients. The albumin levels indicated that a substantial fraction (range: 0.16 0.50) of the resected brain tumor specimens consisted of blood. As the plasma concentration of teniposide during the first hours after infusion is high, the major part of the drug measured in the tumor specimens collected within 2.5 h after drug administration originated from the blood compartment. At 24 h after drug administration, when the plasma level of teniposide had declined to approximately 0.20 microgram/ml, we could discern a real tissue uptake of teniposide ranging from 0.15-0.27 microgram/g wet tissue weight in the resected tumor. Although the number of patients in this study is small, this work clearly illustrates that an accurate determination of the tissue concentration of teniposide is hindered by the high concurrent plasma levels. It is therefore essential that future tissue distribution studies also include a suitable procedure that establishes the contribution of drug originating from the blood compartment. PMID- 9225952 TI - Pharmacokinetics and clinical impact of all-trans retinoic acid in metastatic breast cancer: a phase II trial. AB - PURPOSE: The purpose of this trial was to evaluate tumor cytoreduction by all trans retinoic acid (ATRA) in patients with metastatic breast cancer and to characterize the initial pharmacokinetics of this agent. METHODS: The study was a single institution, phase II study. The treatment regimen consisted of ATRA administered orally at a dose of 50 mg/m2 three times a day for 14 consecutive day of a 21-day cycle. Cycles were repeated until disease progression, unacceptable toxicity or patient withdrawal. Plasma samples were obtained following the first dose of ATRA for pharmacokinetic analysis. RESULTS: A total of 17 patients with metastatic breast cancer were enrolled in the study, and 14 completed at least one cycle of therapy and were evaluable for response. One patient achieved a partial response in soft tissue of 4 months duration. Three patients had stable disease for 4, 2, and 2 months duration. The remainder had progressive disease. ATRA was reasonably well tolerated. Pharmacokinetic analysis revealed a high degree of interpatient variability in systemic exposure following the initial dose of ATRA. CONCLUSIONS: We conclude, that in the dose and schedule tested, ATRA does not have significant activity in patients with hormone refractory, metastatic breast cancer. Future studies should focus on more intensive investigation of those individuals with very high or low ATRA initial systemic exposure in the hope of expanding our understanding of ATRA's clinical pharmacology, ultimately leading to improved efficacy. PMID- 9225953 TI - Measurement of nitrobenzylthioinosine in plasma and erythrocytes: a pharmacokinetic study in mice. AB - PURPOSE: Nitrobenzylthioinosine (NBMPR), a potent inhibitor of nucleoside transport in many cell types, modulates the in vivo disposition of several cytotoxic nucleoside analogs. In this study, a radioligand binding assay was developed for measurement of the NBMPR content of plasma and erythrocytes. METHODS: The assay was based on the competition between NBMPR and [3H]NBMPR for high-affinity sites on human erythrocytes membranes. With this assay, we followed in mice changes in the NBMPR content of blood plasma and erythrocytes, following the intraperitoneal injection of the disodium salt of NBMPR 5'-monophosphate (NBMPR-P), a prodrug form of NBMPR. RESULTS: The radioligand binding assay was able to measure precisely as little as 2.5 pmol of NBMPR, allowing the direct determination of NBMPR concentrations in plasma as low as 16 nM. As few as 8 x 10(3) molecules of NBMPR per cell could be determined in erythrocytes. The NBMPR content of plasma from mice injected with NBMPR-P was maximal at about 20 min after injection and declined to < 0.2% of the peak value by 10 h. Erythrocyte associated NBMPR was also maximal at 20 min, and declined to 11% of the peak value by 10 h after injection. Time courses for the disappearance of NBMPR from plasma and erythrocytes were monoexponential and yielded half-life values of 0.39 h and 0.68 h, respectively, an apparent volume of distribution of 0.61 l/kg, and a clearance of 1.1 l/h per kg. CONCLUSIONS: The radioligand binding assay is a sensitive and facile method for monitoring NBMPR concentrations in mammalian plasma and tissue extracts. PMID- 9225955 TI - A feasibility study of 1-h paclitaxel infusion in patients with solid tumors. AB - The optimal schedule for paclitaxel administration has not yet been determined. This phase I/II study was carried out to evaluate the safety of paclitaxel administration by 1-h infusion in the outpatient setting. A total of 43 patients with advanced pretreated malignancies (18 breast, 18 ovarian, and 7 non-small cell lung cancers) received at least 2 cycles of paclitaxel given at 175 mg/ m2 in a single dose by 1-h i.v. infusion. This protocol was repeated every 21 days. All patients were premedicated as follows: promethazine given i.m. at 50 mg, dexamethasone given at 16 mg in 250 ml normal saline by i.v. infusion for 20 min and ranitidine given i.v. at 50 mg in 250 ml normal saline over 15 min, all premedication being carried out 1 h before the paclitaxel infusion. In a total of 156 cycles, only 1 patient presented with a hypersensitivity reaction (grade 2 urticaria in 1 cycle) and another patient developed transient facial flushing (in 1 cycle: this was resolved by slowing of the infusion rate) on this schedule of paclitaxel administration. Other adverse side effects were usually mild and well tolerated. Alopecia was universal; myelosuppression was uncommon because our patients were supported with granulocyte colony-stimulating factor (G-CSF, lenograstim) given at 34 IU/day in the presence of a neutrophil count of < 500 microliters; neutropenia was seen in 50/156 (32%) cycles and was mild. Neurotoxicity was the most serious adverse effect, and all patients experienced mild to severe neuro-muscular toxicity, mainly in the form of peripheral sensorimotor neuropathy and myalgias. In conclusion, 1-h paclitaxel administration is safe and reduces the duration of treatment, making its use more convenient and easy in the outpatient setting. A prospective comparison of 1-h versus 3-h paclitaxel infusion in terms of efficacy and toxicity is the subject of our current randomized study. PMID- 9225954 TI - A phase II study of weekly high-dose cisplatin combined with oral etoposide in advanced non-small-cell lung cancer. AB - As a dose-response relationship has been suggested for cisplatin, it appeared attractive to explore high-dose-intensity regimens in non-small-cell lung cancer. In a phase I study of weekly administration of cisplatin combined with oral etoposide we achieved a cisplatin dose intensity of 52.5-60 mg/m2 per week in most patients. We subsequently explored this regimen in advanced non-small-cell lung cancer. Patients were treated with cisplatin infused at 70 mg/m2 on days 1, 8, 15 and 29, 36, 43 in combination with oral etoposide given at 50 mg on days 1 15 and 29-43. Patients showing stable disease or a better response were continued on treatment with oral etoposide given at 50 mg/m2 per day on days 1-21 every 28 days for a maximum of four cycles. In all, 22 patients with stage III disease and 31 patients with stage IV disease entered the study. The median number of cisplatin administration was 6 per patient; 17 patients reached the planned cisplatin dose intensity of 60 mg/m2 per week, 11 patients achieved 52.5 mg/m2 per week, and 7 patients reached 47 mg/m2 per week. Overall, 11 of 21 stage III patients had a partial response [response rate 51%, 95% confidence interval (CI) 36-81%], as did 9 of 28 patients with stage IV disease (32%; 95% CI 15-49%). Toxicity was mainly hematologic, with leukocytopenia being the most frequent cause of treatment delay. Nephrotoxicity of grade 1 was observed in seven patients. Two patients developed clinical hearing loss. With this schedule a high median cisplatin dose intensity of 52.5-60 mg/m2 per week was reached. The 51% response rate achieved in stage III disease makes this schedule attractive for further exploration; however, it is not recommended for routine use in stage IV disease. PMID- 9225956 TI - Protective role of metallothionein in renal toxicity of cisplatinum. AB - To elucidate the protective role of metallothionein (MT) in the prevention of cisplatinum (cis-DDP) toxicity, we investigated the lethal and renal toxicities caused by cis-DDP in MT-null transgenic mice in comparison with wild-type control mice, and examined the effect of pretreatment with bismuth nitrate or zinc sulfate on the cis-DDP nephrotoxicity. The MT-null mice were of mixed 129 Ola and C57BL/6 genetic background. Since no differences in cis-DDP nephrotoxicity were observed between these strains, C57BL/6J mice were used as the wild-type control. The basal MT levels in the kidneys were negligible in the MT-null mice and 2.93 +/- 0.77 micrograms/g tissue in the C57BL/6J mice. In terms of both the lethal and renal toxicities of cis-DDP, MT-null mice were far more sensitive than C57BL/6J mice. Preinduction of renal MT synthesis by administration of bismuth nitrate or zinc sulfate protected C57BL/6J mice from cis-DDP nephrotoxicity. In the case of MT-null mice, however, renal MT could not be induced by pretreatment with these metal compounds, and renal toxicity of cis-DDP was not prevented by this pretreatment. These results suggest that MT is an important factor with the potential to suppress the development of cis-DDP toxicity. PMID- 9225957 TI - Adriamycin-induced histamine release from heart tissue in vitro. AB - It has been proven that the anthracyclines induce an important, noncytotoxic histamine release from rat peritoneal mast cells. As mast cells derived from different tissues exhibit marked heterogeneity, the effect of Adriamycin in comparison with other antineoplastic agents was tested on fragments of the right heart auricle, which contain a great number of mast cells. In this experimental model, Adriamycin induced a dose-dependent histamine release that was significantly limited by the antiexocytotic drug sodium cromoglycate. The antineoplastic agents cisplatin and 5-fluorouracil, in contrast, did not provoke any comparable histamine release. In the formulation employed in clinical settings, paclitaxel was also capable of inducing a histamine release comparable with that of Adriamycin; the exocytotic activity, however, was also evident when the tissue fragments were treated with Cremophor EL alone, without the addition of paclitaxel, whereas treatment of samples with paclitaxel dissolved in ethanol did not induce any releasing action. These data thus suggest that the secretory activity should be ascribed to the solvent Cremophor EL and not to paclitaxel. The release of histamine induced by paclitaxel in Cremophor EL/ethanol was also limited by sodium cromoglycate. These results again indicate that histamine release from mast cells derived not only from the peritoneal cavity but also from the cardiac tissue could play a role in the cardiotoxicity of anthracyclines and of paclitaxel in the clinically employed formulation. PMID- 9225958 TI - Methotrexate increases red blood cell concentrations of 6-methylmercaptopurine ribonucleotide in rats in vivo. AB - PURPOSE: To elucidate the effect of methotrexate (MTX) on 6-mercaptopurine (6-MP) metabolism in rats. METHODS: Fourteen rats were given 6-MP 20 mg/kg daily for 7 days. Seven of the rats were also given MTX 20 mg/kg on days 5 and 7. Blood samples were obtained from all rats on days 0.5 and 8, and red blood cell (RBC) lysates were analysed for thiopurine methyltransferase (TPMT) activity and the concentration of methylated 6-MP metabolites [methyl mercaptopurine ribonucleotides (MMPRP)] and 6-thioguanine nucleotides (6-TGN). RESULTS: The concentration of MMPRP increased 2.4 times from day 5 to day 8 in RBCs from rats given MTX in addition to 6-MP, as against 1.2 times in rats given 6-MP alone (P = 0.003). 6-TGN levels increased and TPMT activity decreased from day 5 to day 8, with no difference between the 6-MP and the 6-MP plus MTX groups. CONCLUSIONS: Single bolus doses of MTX increase the concentration of MMPRP in rats given daily s.c. doses of 6-MP, with no effect on 6-TGN concentration or TPMT activity. PMID- 9225959 TI - Chemotherapy in oesophageal cancer. PMID- 9225960 TI - Alternative formulations of paclitaxel. AB - Paclitaxel, a novel antitumour agent, is active clinically against advanced ovarian and breast cancer and under investigation for various other cancers. One of the problems associated with the intravenous administration of paclitaxel is its low solubility in water. The current pharmaceutical formulation consists of a 1:1 (v/v) mixture of ethanol and Cremophor EL. This formulation, however, has been demonstrated to cause some severe hypersensitivity reactions. Therefore the development of a safer intravenous formulation devoid of Cremophor EL is an important investigational issue. This review deals with some of the most promising formulation alternatives. PMID- 9225961 TI - Chimeric T-cell receptors: highly specific tools to target cytotoxic T lymphocytes to tumour cells. PMID- 9225962 TI - Growth factors in human ovarian cancer. PMID- 9225963 TI - Analysis of structural and numerical chromosome abnormalities in sperm of normal men and carriers of constitutional chromosome aberrations. A review. AB - Sperm chromosome analysis offers the opportunity to gather information about the origin of chromosome aberrations in human germ cells. Over the last 20 years more than 20,000 sperm chromosome complements from normal donors and almost 6000 spermatozoa from men with constitutional chromosome aberrations (inversions, translocations) have been analyzed for structural and numerical chromosome abnormalities, as well as for segregation of the constitutional chromosome aberrations after the sperm had penetrated hamster oocytes. On the other hand, it took only 6 years to screen more than 3 million mature spermatozoa from healthy probands for disomy rates of 20 autosomes (chromosomes 19 and 22 not evaluated) and the sex chromosomes, and for diploidy rates by in situ hybridization techniques. In the present paper the results arising from both methods are compiled and compared. PMID- 9225964 TI - Two newly identified mutations (Thr233Ile and Leu152Met) in partially adenosine deaminase-deficient (ADA-) individuals that result in differing biochemical and metabolic phenotypes. AB - Deficiency of adenosine deaminase (ADA-) results in autosomal recessive immunodeficiency disease of varying severity. Partial ADA- [ADA deficiency in erythrocytes (RBCs) but substantial ADA in non-RBCs] has also been identified, primarily by population screening of healthy adults in Africa and newborns in New York State. Normal immune function and/or minimal elevations of toxic metabolites in childhood suggested that partial ADA deficiency was benign and therefore that six mutations identified in partially ADA-deficient newborns and expressing 8-80% of normal ADA in non-RBCs were not pathogenic. However, the lowest activity mutation (Arg211Cys) has now been reported in patients with adult-onset immunodeficiency. We have now molecularly and biochemically studied two additional individuals whom we found to represent opposite ends of the spectrum of partial ADA deficiency as to biochemical abnormalities and age of ascertainment. Homozygosity for a newly identified Leu152Met mutation expressing considerably less activity than the pathogenic Arg211Cys mutation was found in a currently healthy 10-year-old Afghanistani child (ascertained at birth). He had the highest accumulation of the metabolite dATP among 13 partially ADA-deficient patients studied, but considerably lower than in those with immunodeficiency. Homozygosity for a newly identified Thr233Ile mutation expressing somewhat greater ADA activity than Arg211Cys was found in a healthy young adult Kung individual, associated with very low metabolite concentrations. Biochemical findings and a family history suggestive of immunodeficiency in prior offspring support the idea that the Leu152Met mutation could result in disease in homozygous individuals challenged by severe environmental insult or in heterozygosity with a null mutation. The pathogenicity of the Thr233Ile mutation, as well as a previously described Ala215Thr mutation with relatively lower activity is less likely but will only be determined by long-term observation of individuals carrying these mutations. Although, in contrast to other partial mutations, neither of these two mutations are at CpG hot spots, the frequency of CpG mutations remains high for partial mutations but is also similarly high in ADA- immunodeficient patients (5/8 vs 12/21). PMID- 9225965 TI - Genetic variations at the T cell receptor gamma locus in circulating peripheral blood mononuclear cells of clinically categorised leprosy patients. AB - The allelic polymorphisms at exon 3 and exon 2 of the T cell receptor (TCR) C gamma 2 (TRGC2) gene, generating 18-kb and 5.4-kb HindIII fragments, respectively, were found to be more frequent in multibacillary leprosy patients than in the controls (P < 0.005 and P < 0.001, respectively) when screened with the IDP2.11 probe. The frequencies of heterozygotes for the 18-kb allele and homozygotes for the 5.4-kb allele were found to be significantly higher in the multibacillary patients than in the controls (P < 0.001). Interestingly, the 8.0 kb allele, originating from the triplication of exon 2 of C gamma 2, was observed exclusively in the paucibacillary leprosy patients. Further, when DNA samples were screened with the pH60 probe for the HindIII RFLP at the TCR J gamma 2 (TRGJ2) gene segment, the 2.1-kb allele was again more prevalent in leprosy patients with the multibacillary form of the disease than in the paucibacillary patients and the controls (P < 0.025). The frequency of homozygotes for the 2.1 kb allele was also significantly higher in the multibacillary patients than in the paucibacillary patients (P < 0.010) and the controls (P < 0.025). A significant difference was observed in the frequencies of detectable rearrangements involving the V gamma 7/8 and V gamma 9 gene segments at the gamma locus between circulating peripheral blood mononuclear cells of the multibacillary leprosy patients and the controls. These rearrangements were detected less frequently in the multibacillary patients (P < 0.001 for V gamma 7/8 and P < 0.005 for V gamma 9). PMID- 9225966 TI - Search for the optimal fetal cell antibody: results of immunophenotyping studies using flow cytometry. AB - Fetal nucleated cells circulating in maternal peripheral blood are a noninvasive source of fetal DNA for prenatal genetic diagnoses. The successful isolation of fetal cells from maternal blood depends upon identification of differences between fetal and maternal cell surface antigen expression. To our best knowledge, a monoclonal antibody that binds only fetal blood cells has not yet been identified. We studied antigens recognized by six different monoclonal antibodies for their biologic expression on fetal blood cells as a function of gestational age, and compared their ability to bind fetal but not maternal cells. The results suggest a relationship between gestational age and nucleated cell surface antigen expression. The monoclonal antibodies FB3-2, H3-3, CD71 and 2 6B/6 are suitable reagents for first or early second trimester fetal cell isolation, although FB3-2 and H3-3 are more specific for fetal cells due to significantly lower expression of these antigens on maternal mononuclear cells. The observation that samples from fetuses with chromosome abnormalities or multiple structural anomalies express higher levels of these antigens indicates that these reagents will potentiate the detection of abnormal fetal cells in maternal blood samples. PMID- 9225968 TI - Multiple identification of a particular type of hereditary C1q deficiency in the Turkish population: review of the cases and additional genetic and functional analysis. AB - Complete selective deficiencies of the complement component C1q are rare genetic disorders that are associated with recurrent infections and a high prevalence of lupus erythematosus-like symptoms. All C1q deficiencies studied at the genetic level revealed single-base mutations leading to termination codons, frameshifts or amino acid exchanges and these were thought to be responsible for the defects as no other aberrations were found. One particular mutation, leading to a stop codon in the C1qA gene, was first identified in members of a Gypsy family from the Slovak Republic. The same mutation has been found in all cases of C1q deficiency from Turkey that have been investigated. Here we present the results of genetic analysis of the C1q genes from three families and give information on further C1q-deficient members of two families that have not been reported elsewhere. Reviewing all cases of C1q deficiency from Turkey prompted us to hypothesize that one particular defective allele is present in the population of southeast Europe and Turkey. With a novel polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis and allele-specific PCR we are able to detect even asymptomatic, heterozygous carriers of the mutation, which will enable genetic counseling of the affected individuals. PMID- 9225967 TI - Association of polymorphism at COL3A and CTLA4 loci on chromosome 2q31-33 with the clinical phenotype and in-vitro CMI status in healthy and leprosy subjects: a preliminary study. AB - Two genetic loci, viz. COL3A and CTLA4, located within the chromosome 2q31-33 region in the vicinity of the proposed syntenic site of the mouse "Bcg" locus were genotyped by the polymerase chain reaction in leprosy patients and healthy individuals. All the subjects studied were assessed as in-vitro responders/non responders to mycobacterial antigens. Simple sequence length polymorphism analysis revealed five (236 to 312 bp) and eight (84 to 120 bp) allelomorphs for COL3A and CTLA4, respectively. Our preliminary analysis showed a significant association between the 250-bp COL3A allelomorph in the homozygous condition and the multibacillary form of leprosy (P < 0.05: relative risk = 5.5). Another allelic (312 bp) variant of COL3A was significantly correlated with non responsiveness to M. leprae antigens in vitro (P < 0.01). The 104-bp allelomorph of CTLA4 was not observed in any of the 25 cases of leprosy. This absence was statistically significant (P < 0.05) when compared with normal healthy controls and depicted a high relative risk (RR = 25.83). An additional observation of the predominance of a unique 84-bp CTLA4/CTLA4-like allelomorph was observed in the Indian subjects studied. PMID- 9225969 TI - Molecular basis of the apolipoprotein H (beta 2-glycoprotein I) protein polymorphism. AB - Apolipoprotein H (apoH, protein; APOH, gene) is considered to be an essential cofactor for the binding of certain antiphospholipid autoantibodies to anionic phospholipids. APOH exhibits a genetically determined structural polymorphism due to the presence of three common alleles (APOH*1, APOH*2 and APOH*3) detectable by isoelectric focusing (IEF) and immunoblotting. The APOH*3 allele can be further characterized into two subtypes, APOH*3w and APOH*3B, based upon its reactivity with monoclonal antibody 3D11. In this study we have determined the molecular basis of the APOH protein polymorphism and its distribution in three large U.S. population samples comprising 661 non-Hispanic whites, 444 Hispanics and 422 blacks. By direct DNA sequencing of PCR amplified fragments corresponding to the eight APOH exons, we identified two missense mutations that correspond to the APOH*1 and APOH*3w alleles. A missense mutation (G-->A) in exon 3, which alters amino acid Ser to Asn at codon 88 and creates a restriction site for TSP509 I, was present in all APOH*1 allele carriers. A second missense mutation (G-->C) at codon 316 in exon 8, which replaces amino acid Trp with Ser and creates a restriction site for BSTBI, was present in all APOH*3w carriers. The distribution of the Ser 88 Asn and Trp 316 Ser mutations was significantly different between the three racial groups. The frequency of the Asn-88 allele was 0.011, 0.043, and 0.056 in blacks. Hispanics and non-Hispanic whites, respectively. While the Ser 316 allele was observed sporadically in blacks (0.008), it was present at a polymorphic frequency in Hispanics (0.027) and non-Hispanic whites (0.059). The identification of the molecular basis of the APOH protein polymorphism will help to elucidate the structural-functional relationship of apoH in the production of antiphospholipid autoantibodies. PMID- 9225970 TI - Genetic investigation of the porphobilinogen deaminase gene in Swedish acute intermittent porphyria families. AB - A total of 12 mutations associated with acute intermittent porphyria (AIP) have been detected in the porphobilinogen deaminase gene in Swedish AIP families. Three of them are newly discovered and unique to the Swedish population: a splice mutation in intron 6 (int6+1), a missense mutation in exon 11 (Gly216Asp) and a TG deletion in exon 14. PMID- 9225971 TI - Mutational analysis and expression studies of the neurofibromatosis type 2 (NF2) gene in a patient with a ring chromosome 22 and NF2. AB - The case of a seriously disabled and retarded female patient with neurofibromatosis type 2 (NF2) is reported. She suffered from bilateral vestibular schwannomas, multiple intracranial meningiomas and neurinomas. The constitutional karyotype of the patient was 46, XX, r(22)/45,XX,-22. A constitutional G to A transition in the proximal 3' untranslated region of isoforms 1 and 2 was identified in the patient's NF2 gene and shown not to affect differential splicing or mRNA stability. The instability of the ring chromosome 22 with the associated loss of tumor suppressor genes on chromosome 22, in particular the loss of the NF2 gene, are assumed to have caused multiple tumorigenesis in this patient. PMID- 9225973 TI - Human cationic amino acid transporter gene hCAT-2 is assigned to 8p22 but is not the causative gene in lysinuric protein intolerance. AB - Lysinuric protein intolerance (LPI) is a recessively inherited amino acid disorder characterized by defective efflux of cationic amino acids at the basolateral membrane of the intestinal and renal tubular epithelium. Recently, cDNAs encoding the related proteins hCAT-2A and hCAT-2B have been cloned. These two carrier proteins are most likely to product of the same gene, hCAT-2. Using the hCAT-2B cDNA, we assigned the hCAT-2 gene to chromosome 8p22. Furthermore, by linkage analysis in Finnish LPI families, we ruled out that hCAT-2B is involved in LPI disease. PMID- 9225972 TI - Genetic fine localization of the beta-glucocerebrosidase (GBA) and prosaposin (PSAP) genes: implications for Gaucher disease. AB - Mutations in the glucocerebrosidase (GBA) and prosaposin (PSAP) genes are responsible for Gaucher disease, the most prevalent sphingolipidosis. Somatic cell hybrid analysis and in situ hybridization experiments have localized the GBA gene to 1q21 and the PSAP gene to 10q21-q22. We performed pairwise and multi point linkage analyses between the two genes and several highly polymorphic markers from the Genethon human linkage map. Our results show that six markers cosegregate with the GBA gene (Zmax = 8.73 at theta = 0.00 for marker D1S2714) and define a 3.2-cM interval between D1S305 and D1S2624 as the most probable location for the gene. Three of these markers (D1S2777, D1S303, and D1S2140), as well as the gene encoding pyruvate kinase (PKLR), are contained in a single YAC clone together with the GBA gene. A new polymorphism was identified within the PSAP gene (C16045T) and used for linkage studies. The multi-point analysis places the gene in a 9.8-cM interval between D10S1688 and D10S607. The fine localization of these genes provides a useful tool for cosegregation analysis, indirect molecular diagnosis, and population genetic studies. PMID- 9225975 TI - Identification of novel 'expressed sequence tags' within the FHIT gene locus in human chromosome region 3p14.2. AB - Losses of genetic material within human chromosome regions (HCR) 3p12-p14 and 3p21-p22 are observed in various neoplasias, suggesting tumor suppressor gene (TSG) loci within these regions. HCR 3p14 is particularly interesting as it contains the t(3;8) translocation breakpoint of a hereditary renal cell carcinoma, the FRA3B fragile site, and DNA markers deleted in several types of human cancer. We here report on the identification of five novel 'expressed sequence tags' (ESTs) within 3p14.2 which map proximal to exon 9 of the candidate TSG, FHIT. These ESTs may be valuable for elucidation of the supposed TSG content in 3p14.2. PMID- 9225976 TI - Discriminating between allelic and interlocus differences among human immunoglobulin VH4 sequences by analyzing single spermatozoa. AB - To address the challenging issue of distinguishing allelic and interlocus differences among repetitive sequences, human immunoglobulin VH4 loci in the parental haplotypes of 13 donors were determined by analyzing single spermatozoa. VH4 sequences detected among these donors were assigned to their corresponding loci based on the fact that allelic sequences usually segregate into different gametes. Four out of the ten VH4 loci were shown to contain null alleles that are undetectable with diploid materials. The distribution of the allelic variation within the analyzed regions at the VH4 loci is highly biased. PMID- 9225974 TI - The G4 gene is duplicated in 44% of human immunoglobulin heavy chain constant region haplotypes. AB - The structure of the human immunoglobulin heavy chain constant region (IGHC), on chromosome 14q32, comprises nine CH genes and two pseudogenes, all originating from multiple duplication events. Continuing evolution of the region is demonstrated by the finding of various types of duplicated and deleted haplotypes, which together add up to 6%. Here we provide molecular and genetic evidence that the G4 gene is duplicated in 44% of IGHC haplotypes in the Italian population. The duplication spans about 20 kb of genomic DNA and probably originated through unequal crossing over. Refined characterisation of the genomic region downstream from the G4 gene improves our knowledge of the evolutionary history of CH genes. PMID- 9225977 TI - A double mutant LDL receptor allele in a cypriot family with heterozygous familial hypercholesterolemia. AB - Two novel mutations Q363X and D365E were identified in the low-density lipoprotein receptor gene in a Cypriot patient with heterozygous familial hypercholesterolemia. Restriction enzyme analysis of the index case and seven of her family members, by using AvaII and PvuII respectively, demonstrated that the two exon 8 mutations are transmitted in cis within the family. The disease phenotype is probably caused by the stop-363 mutation; this would result in a truncated protein that would probably be rapidly degraded in the extracellular space. PMID- 9225978 TI - An unusual combined insertion/deletion polymorphism in intron 10 of the human complement C6 gene. AB - Investigation of intron 10 of the human complement C6 gene revealed an unusual combined insertion/deletion polymorphism at position 493: the subsequent 6 bp is deleted and is substituted by a different 26 bp, giving a net gain of 20 bp. The variant shows autosomal co-dominant inheritance. The 26 bp insertion is homologous to a human endogenous retrovirus-type sequence and could tentatively be ascribed to a retroposon. Alternatively, the presence of three copies of a 5 bp direct repeat, an 8 bp palindrome and a 12 bp split symmetrical element could suggest an endogenous, sequence-mediated mutational process. Polymorphisms of this type are extremely rare, although there are several examples of such mutations causing disease. PMID- 9225979 TI - Regional distribution of beta-thalassemia mutations in India. AB - We have characterized the mutations in 1050 carriers of the beta-thalassemia gene and analyzed their regional distribution in India. The majority of beta thalassemia carriers were migrants from Pakistan and their pattern of mutations differed from the rest. The frequency of the 619-bp deletion was 33.3% among the migrants from Pakistan, 8-17% in the northern states, and less than 5% in the other states. Among non-migrant subjects, the predominant mutation was IVS-I-5 (G ->C), varying from 85% in the southern states and 66-70% in the eastern states to 47-60% in the northern states. The mutation IVS-I-1 (G-->T) was observed at high frequency among the migrants from Pakistan (26.2%), but with very low/zero frequency in the other states. Mutations at codons 8/9 (+G) and codons 41/42 ( CTTT) were distributed in all regions of India with a frequency varying from 3% to 15%. Only eight of 12 published rare mutations were observed in subjects from different parts of India. Mutations of codon 5 (-CT) and codons 47/48 (+ATCT) were found exclusively in migrants from Pakistan, and mutation -88 (C-->T) was detected only in subjects from Punjab, Haryana, and Uttar Pradesh. Using the amplification refractory mutation system technique, mutations were successfully identified in 98.2% of subjects. Overall, 91.8% of the subjects had one of the five commonest mutations [IVS-I-5 (G-->C), 34.1%; 619-bp deletion, 21.0%; IVS-I-1 (G-->T) 15.8%; codons 8/9 (+G), 12.1%, and codons 41/42 (-CTTT), 8.7%], 5.9% of the subjects had a less common mutation, while 1.8% of the carriers remained uncharacterized. The application of this knowledge has helped to successfully establish a program of genetic counselling and prenatal diagnosis of beta thalassemia in order to reduce the burden of this disease in India. PMID- 9225981 TI - Mutation and haplotype analyses of the Werner's syndrome gene based on its genomic structure: genetic epidemiology in the Japanese population. AB - The correlation between mutations in the Werner's syndrome (WRN) gene and the haplotypes of surrounding markers was studied in Japanese patients. We have elucidated the genomic structure of WRN helicase, and found five additional mutations, designated mutations 6-10. Mutations 4 and 6 were found to be the two major mutations in this population; these mutations comprised 50.8% and 17.5%, respectively, of the total in a sample of 126 apparently unrelated chromosomes. Almost all the patients homozygous for mutation 4 shared a haplotype around the WRN gene, consistent with the view that they are derived from a single ancestor. This important advantage demonstrated in the identification of the WRN gene suggests that the Japanese present a unique population for the cloning of other disease genes. The conserved haplotype was observed across 19 loci, extending a distance estimated to be more than 1.4 Mbp around the WRN gene. This haplotype is rare among random Japanese individuals. Unexpectedly, all the nine patients homozygous for mutation 6 shared a haplotype that was identical to this haplotype at 18 of these 19 markers. These results suggest that mutations 4 and 6 arose independently in almost identical rare haplotypes. The remaining mutations (1, 5, 7, 8, 9, and 10) occurred rarely, and were each associated with different haplotypes. PMID- 9225980 TI - cDNAs with long CAG trinucleotide repeats from human brain. AB - Twelve diseases, most with neuropsychiatric features, arise from trinucleotide repeat expansion mutations. Expansion mutations may also cause a number of other disorders, including several additional forms of spinocerebellar ataxia, bipolar affective disorder, schizophrenia, and autism. To obtain candiate genes for these disorders, cDNA libraries from adult and fetal human brain were screened at high stringency for clones containing CAG repeats. Nineteen cDNAs were isolated and mapped to chromosomes 1, 2, 4, 6, 7, 8, 9, 12, 16, 19, 20, and X. The clones contain between 4 and 17 consecutive CAG, CTG, TCG, or GCA triplets. Clone H44 encodes 40 consecutive glutamines, more than any other entry in the nonredundant GenBank protein database and well within the range that causes neuronal degeneration in several of the glutamine expansion diseases. Eight cDNAs encode 15 or more consecutive glutamine residues, suggesting that the gene products may function as transcription factors, with a potential role in the regulation of neurodevelopment or neuroplasticity. In particular, the conceptual translation of clone CTG3a contains 18 consecutive glutamines and is 45% identical to the C terminal 306 residues of the mouse numb gene product. These genes are therefore candidates for diseases featuring anticipation, neurodegeneration, or abnormalities of neurodevelopment. PMID- 9225982 TI - Expanded CAG repeats in spinocerebellar ataxia (SCA1) segregate with distinct haplotypes in South african families. AB - The autosomal dominant late onset spinocerebellar ataxias (SCAs) are genetically heterogeneous. Three genes, SCA1 on 6p, SCA2 on 12q and MJD1 on 14q, have been isolated for SCA1, SCA2 and Machado-Joseph disease (MJD), respectively. In these three autosomal dominant disorders the mutation is an expanded CAG repeat. Evidence for heterogeneity in families not linked to the SCA1, SCA2 and MJD loci is provided by the mapping of SCA loci to chromosomes 16q, 11cen and 3p. A total of 14 South African kindreds and 22 sporadic individuals with SCA were investigated for the expanded SCA1 and MJD repeats. None of the families nor the sporadic individuals showed expansion of the MJD repeat. Expanded SCA1 and CAG repeats were found to cosegregate with the disorder in six of the families tested and were also observed in one sporadic individual with a negative family history of SCA. The use of the microsatellite markers D6S260, D6S89 and D6S274 provided evidence that the expanded SCA1 repeats segregated with three distinct haplotypes in the six families. Use of the highly polymorphic tightly linked microsatellite markers is still important as this stage, particularly where this coincides with the possibility of a homozygous genotype with the trinucleotide repeat marker. Importantly, our molecular findings indicate: (1) an absence of MJD expanded repeats underlying SCA; (2) the major disease in this group is due to mutations in the SCA1 gene; and (3) the familial disorder in the majority population group (i.e. mixed ancestry) in the Western Cape region of South Africa is most likely to be the result of two distinct founder events. PMID- 9225983 TI - Analysis of the variation in chromosome size among diverse human populations by bivariate flow karyotyping. AB - Chromosomes sampled from seven human populations were analyzed by flow cytometry to survey normal variation in chromosome size. The populations include two African Pygmy groups, two Amerindian tribes, Druze, Khmer Cambodians, and Melanesians. Mitotic chromosomes were isolated from cultured cells and stained with Hoechst 33,258 and chromomycin A3. The relative DNA content and base-pair composition of each homolog was quantified by bivariate flow karyotyping. Significant variation in DNA content, ranging from 10-40%, was observed for chromosomes 1, 13-16, 19, 21, 22, and Y. The measurements for each population appeared to be a random sampling of the total set of 33 individuals for the majority of chromosomes. A few significant differences in the distributions of chromosomal DNA content were observed among the populations, however. The data, when combined with an earlier study of 33 unrelated individuals of unknown ethnic origin, provide a good representation of the variation in chromosome size among humans. PMID- 9225985 TI - A continuing enigma: the role of cells of macrophage lineage in the development of HIV disease. PMID- 9225984 TI - Microsatellite polymorphism in the human heme oxygenase-1 gene promoter and its application in association studies with Alzheimer and Parkinson disease. AB - Oxidative stress has been suggested to be involved in the pathogenesis of neurodegenerative diseases, such as Alzheimer disease (AD) and Parkinson disease (PD). Heme oxygenase-1 (HO-1), a key enzyme in heme catabolism, also functions as an antioxidant enzyme. Here, we show that a (GT)n repeat in the human HO-1 gene promoter region is highly polymorphic, although no particular alleles are associated with AD or PD. This newly identified genetic marker should allow us to study the possible involvement of HO-1 in certain human diseases. PMID- 9225986 TI - Macrophage tropism: fact or fiction? AB - Primary HIV-1 isolates can be distinguished by phenotypic qualities such as the ability to productively infect cells of established CD4-positive lines and to induce syncytia in MT-2 cells. Such viral phenotypes have also been reported to confer host cell specificity. It is perceived that primary isolates with the syncytium-inducing phenotype (SI or rapid/high) are T cell tropic and are therefore unable to infect primary cells of the monocyte/macrophage lineage. However, we have consistently found that these isolates are as capable of establishing infection in monocyte-derived macrophages (MDM) as the monocytotropic, non-syncytium-inducing variants (NSI or slow/low). It is known that differentiation, activation, and proliferation of human monocytes are affected by both isolation methods and culture conditions. Therefore, to test whether our inability to discriminate macrophage tropic HIV-1 isolates could be explained by differences in culturing techniques, we isolated monocytes by elutriation or short-term adherence and allowed the cells to mature and differentiate in either the presence or absence of autologous lymphocytes. After removal of nonadherent cells, MDM were infected with a panel of SI and NSI primary HIV-1 isolates. MDM were susceptible to infection by the SI as well as the NSI isolates, regardless of whether or not the cells were allowed to mature in the presence of autologous lymphocytes. However, MDM matured in the presence of autologous lymphocytes replicated HIV-1 isolates (both NSI and SI) to a higher titre than MDM matured in the absence of lymphocytes. In light of these findings and recently published reports on HIV-1 phenotype and chemokine receptor usage we believe that the term macrophage-tropic strains of HIV-1 is no longer appropriate. PMID- 9225987 TI - Significance of macrophage tropism of SIV in the macaque model of HIV disease. AB - Microglia, alveolar macrophages, and Langerhans cells are representatives of cells of macrophage lineage that are susceptible to infection with HIV-1 and they play important roles in the pathogenesis of AIDS dementia, lymphoid interstitial pneumonia, and systemic viral invasion from mucosal surfaces, respectively. In contrast, elimination of CD4+ T cells with resultant development of immunosuppression and AIDS is thought to be reflective of the exclusive tropism of the virus for CD4+ T cells. Examination of these concepts in macaques infected with molecularly cloned strains of SIVmac suggested that all strains of the virus are both macrophage- and lymphocyte-tropic and that all aspects of pathogenesis including loss of CD4+ T cells are dependent on infection in both cell types. However, viral clones that caused productive lytic infection in macrophages were less virulent than those which caused persistent nonproductive infection. The former caused subclinical and even immunizing infections, whereas the latter caused activation and productive infection in CD4+ T cells, AIDS, and systemic infection, even after inoculation of the virus on mucosal surfaces. If these findings on SIVmac are relevant to HIV-1 disease, then demonstration that HIV-1 isolates are macrophage-tropic probably does not necessarily correlate with their pathogenic potential. PMID- 9225988 TI - Chemokine receptors and HIV. AB - The discovery that chemokine receptors are the human cofactors required along with CD4 for fusion and infection by HIV has opened new directions in AIDS research on mechanisms of viral entry, tropism, and pathogenesis. A possible mechanism of co-receptor function has been demonstrated that involves the formation of a complex on the cell surface between the HIV-1 envelope, CD4, and the coreceptor. Functional studies indicate that this interaction is structurally complex, that it probably involves multiple domains of the coreceptor, and that different virus isolates interact with coreceptors in distinct ways. Other immunodeficiency viruses including simian immunodeficiency virus and feline immunodeficiency virus also utilize chemokine receptors for entry. The identification of genetic polymorphisms helps explain why some people, with alterations in the CCR5 gene that prevent expression, are protected from HIV-1 infection. The discovery of specific HIV-1 fusion coreceptor molecules has not only provided new insights into the mechanisms of viral entry and tropism, but also led to new avenues of investigation on strategies to block HIV infection. PMID- 9225990 TI - Role of pro-inflammatory cytokines and beta-chemokines in controlling HIV replication. AB - Several members of the cytokine network play an important role in controlling the replication of the human immunodeficiency virus (HIV) in several experimental systems. Their effects can be categorized in the following three functional groups: (1) HIV-inductive cytokines; (2) HIV-suppressive cytokines; (3) cytokines with both activating and inhibiting capacities. Studies on the mechanism of action of these molecules have highlighted the fact that several steps of the retrovirus life cycle, from binding to budding of progeny virions from the infected cell, are affected by cytokines. This general concept has been recently substantiated by the discovery that certain beta-chemokines can act as blockers of viral entry by interfering with HIV co-receptors. Finally, it is important to recognize that cytokines have gone beyond their role as potential pathogenetic or protective endogenous cofactors in HIV replication and disease progression, and are becoming experimental therapeutic agents for HIV disease, best illustrated thus far by the case of interleukin-2. PMID- 9225991 TI - GM-CSF and its effects on replication of HIV-1 in cells of macrophage lineage. AB - Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a hemopoetic growth factor that is a member of the four-helix bundle family of cytokines and growth factors. It regulates the proliferation and differentiation of granulocytes and cells of macrophage lineage from bone marrow progenitor cells, mediating these activities through binding to its receptor. Most studies examining the effects of GM-CSF on HIV-1 replication in primary monocytes and macrophages, and in related cell lines, have demonstrated augmentation of HIV-1 expression in vitro, although some reports have been at variance with these findings. These laboratory-based observations have been confirmed in limited clinical trials. This review outlines the details of these studies and considers mechanisms by which GM-CSF may exert its effects on cells of this lineage. PMID- 9225989 TI - MCP-1 and CCR2 in HIV infection: regulation of agonist and receptor expression. AB - Monocyte chemotactic protein-1 (MCP-1) interacts with the chemokine receptor CCR2. Two CCR2 cDNAs have been described. Sequence analysis as well as Northern blotting and RNase protection with different probes revealed that the CCR2 gene is expressed in activated natural killer (NK) cells and mononuclear phagocytes as a predominant long transcript (3.4 kb) consisting of CCR2B followed by a novel sequence (X), corresponding to an intron in the genome, and by a CCR2A specific portion. The predominant long transcript is polyadenylated and present in the cytoplasm. We found that bacterial products and cytokines affect CCR2 expression. Interleukin-2 (IL-2) augmented CCR2 mRNA in monocytes and NK cells. The augmented migratory capacity of IL-2-activated versus resting NK cells was associated with increased CCR2 transcript levels. Lipopolysaccharide (LPS) and other microbial agents caused a rapid and drastic reduction of CCR2 mRNA levels. The rate of nuclear transcription of CCR2 was not affected by LPS, whereas the mRNA half life was reduced. These results suggest that regulation of receptor expression, in addition to agonist production, is probably a crucial point in the regulation of the chemokine system. Down-regulation of chemokine receptor expression may play a role in the modulation of HIV infection in macrophages by LPS. Levels of MCP-1 were markedly elevated in the cerebrospinal fluid (CSF) but not in blood of HIV infected patients with cytomegalovirus (CMV) encephalitis. The CSF levels of MCP 1 in CMV encephalitis were markedly higher than those found in the CSF of HIV infected patients with or without unrelated neurological diseases. IL-8, the prototype of C-X-C chemokines and RANTES and macrophage inflammatory protein-1 alpha (C-C chemokines) were not substantially increased in the liquor of CMV encephalitis patients. High levels of MCP-1 may underlie monocyte recruitment and tissue damage in CMV encephalitis and may represent a rapid and useful tool in the diagnostic armamentarium for neurological disorders associated with HIV. PMID- 9225992 TI - Induction of cytokines by HIV-1 and its gp120 protein in human peripheral blood monocyte/macrophages and modulation of cytokine response during differentiation. AB - We previously reported that in vitro culture of human peripheral blood monocytes resulted in a time-dependent differentiation into macrophages and in an enhanced capacity for producing certain cytokines [i.e., tumor necrosis factor alpha, interleukin-6 (IL-6), and interferon-beta (IFN-beta)] in response to bacterial lipopolysaccharide (LPS). HIV-1 infection or gp120 treatment of monocyte/macrophages resulted in the induction of low levels of IFN-beta, which were very effective in restricting viral replication in 7-day cultured macrophages but not in freshly isolated cells. This enhanced response of macrophages was due to a higher sensitivity of these cells to the antiviral effect of IFN-beta. Consistent with this finding, 7-day cultured macrophages exhibited higher levels of type I IFN receptors than 1-day cultured monocytes. Treatment of monocyte/macrophages with gp120 also caused a marked increase in IL 10 secretion, regardless of the differentiation state. No IL-12 secretion was detected in monocyte/macrophage cultures treated with gp120 alone. However, consistent IL-12 secretion was found in 7-day cultured macrophages primed with IFN-beta and subsequently stimulated with gp120. Macrophages responded more efficiently than monocytes to the priming effect of IFN-beta for IL-12 production. This was consistent with a stronger antiviral response against vesicular stomatitis virus by these cells as well as with a higher expression of IFN-beta receptors. The finding that the acquisition of the macrophage phenotype is associated with an increased capacity to respond to environmental signals (such as type I and type II IFNs) underlines the importance of the differentiation process for the selection of a certain repertoire of responses that may allow these cells to have important functions in vivo. PMID- 9225993 TI - Intracellular GSH content and HIV replication in human macrophages. AB - In vitro HIV-1 infection induced a significant decrease in intracellular reduced glutathione (GSH) in human macrophages. Such a decrease was observed at the time of infection corresponding to maximum release of virus from infected cells and was not related to cell cytotoxicity. GSH los was not related to its oxidation or leakage through the cell membrane. Inhibition of intracellular GSH synthesis by buthionine sulfoximine (BSO) did not further decrease GSH levels with respect to the decrease caused by HIV alone. However, treatment of macrophages with BSO significantly increased the HIV yield in the supernatant. Exogenous GSH strongly suppressed the production of p24 gag protein as well as the virus infectivity. Previous observations with other RNA and DNA viruses consistently showed that GSH antiviral effect occurred at late stages of virus replication and was related to the selective decrease of specific glycoproteins, such as gp120, which are particularly rich in disulfide bonds. PMID- 9225994 TI - Possible participation of polymorphonuclear cells stimulated by microbial immunomodulators in the dysregulated cytokine patterns of AIDS patients. AB - Macrophages and polymorphonuclear cells (PMN) play a major role as cells primarily responsive to microbial biological response modifiers (BRM). Although much attention has been given to macrophages, PMN have been relatively underinvestigated. We have recently studied the responses of PMN from HIV- and HIV+ subjects after stimulation with a powerful immunomodulatory fraction from the cell wall of Candida albicans (MP-F2) and compared this to bacterial lipopolysaccharide (LPS). Both cytokine patterns and PMN anticandidal activity were investigated. MP-F2, like LPS, was an active inducer of interleukin-8 (IL 8), tumor necrosis factor alpha (TNF-alpha), IL-6, and IL-1 beta production by PMN and monocytes from all subjects. IL-12 was also produced by MP-F2-stimulated PMN in the presence of interferon-gamma (IFN-gamma). PMN from HIV+ subjects showed increased in vitro expression of TNF-alpha and IL-6 genes as determined by semiquantitative reverse transcriptase-polymerase chain reaction. In all subjects, cytokine gene expression was strongly stimulated by MP-F2 or LPS and inhibited by IL-10. Production of IL-6 and TNF-alpha protein (measured by ELISA) was higher in PMN from HIV+ subjects in at least one of the conditions tested (unstimulated or stimulated by LPS or MP-F2). However, the amount of the C-X-C chemokine IL-8 was equal in PMN from HIV- and HIV+ subjects. PMN from HIV+ subjects were at least as active in inhibiting candide growth as PMN from HIV- controls. In both groups PMN were equally stimulated by MP-F2 and LPS. Only in severely neutropenic subjects was there some reduction in the anticandidal activity but not in cytokine responses. When appropriately stimulated by microbial BRM, PMN are active producers of pro-inflammatory and immunomodulatory cytokines. This production is not only totally preserved in HIV+ subjects but may be higher than in PMN from HIV- subjects and may be coupled with an efficient anticandida activity. We suggest that during common bacterial or fungal infections PMN may contribute to the dysregulated production of inflammatory cytokines in AIDS patients. PMID- 9225995 TI - Immune stimulation and HIV-1 viral replication. AB - A biphasic early and late viremia is characteristic of HIV-1 infection. The first increase in circulating viral burden occurs within weeks after infection, before a host immune response, and the second, later peak emerges during the inevitable HIV-1 devastation of immune function. Recently, intermittent bouts of viremia have also been identified in HIV-1-infected individuals and found to be associated with episodes of immune challenge. Vaccinations, exposure to antigens, and infections often induce reversible increases in circulating viral levels, dependent on CD4+ T lymphocyte numbers. However, even with marked losses in CD4+ T cell counts, opportunistic infections appear to trigger a viremic response. In searching for the source of this virus, macrophages in tissues co-infected with opportunistic pathogens have been identified as prodigious producers of HIV-1. Thus, the fountain from which HIV-1 emerges may shift from CD4+ T lymphocytes in early HIV-1 infection to tissue macrophages later in the natural evolution of the disease, as the CD4+ T cells are depleted. Defining the mechanisms of this transitional event in HIV-1 infection may facilitate regulation and therapeutic control of both opportunistic infections and HIV-1. PMID- 9225996 TI - Infection of gastrointestinal tract macrophages by HIV-1. AB - As the largest lymphoid organ and the largest reservoir of macrophages in the body, the gastrointestinal tract mucosa is probably the largest organ reservoir of macrophages infected with HIV-1. To elucidate the biology of HIV-1 infection of intestinal macrophages, we isolated lamina propria macrophages from normal human jejunum by neutral protease digestion, purified the cells by counterflow centrifugal elutriation, and then infected the cells with HIV-1. The lamina propria macrophages were permissive to macrophagetropic isolates of HIV-1 and substantially less permissive to lymphocyte-tropic isolates. Compared with blood monocytes, mucosal macrophages produced 2-3 logs less p24 antigen at peak infection. The reduced level of infection was not due to impaired macrophage viability, reduced CD4 expression, or the isolation procedure. These results confirm that macrophages isolated from the gastrointestinal tract mucosa can support HIV-1 production, albeit at a lower level than blood monocytes. The reduced level of virus production may reflect the unique biology of intestinal lamina propria macrophages. PMID- 9225997 TI - Mechanisms of loss of functional dendritic cells in HIV-1 infection. AB - Dendritic cells (DC) are lost from blood and skin during injection with HIV-1; those remaining show a reduced capacity to stimulate T cell proliferation [S. C. Knight, AIDS 10, 807-817]. Our recent studies investigate mechanisms underlying these effects. DC exposed to HIV-1 vitro can act as targets for cytotoxic T cells, although optimal killing was not obtained until DC were exposed to HIV-1 for 3 days. This cytotoxicity may provide a feedback mechanism by which DC that have presented antigens are removed. However, this effect could also contribute to the reduction in DC during persistent infection. We have also investigated the effect of exposure to HIV-1 on DC function. DC exposed to HIV-1 IIIB virus for 2 h stimulated primary proliferative and cytotoxic T cell responses in vitro; these effects may be similar to those occurring during the early activation of protective antiviral immunity in vivo. After exposure of DC to virus for 5 days, stimulation of allogeneic T cells was reduced. However, a different situation applied when using DC developed from CD34+ cord blood stem cells under the influence of granulocyte-macrophage colony-stimulating factor and tumor necrosis factor alpha that were exposed at 24 h to the same virus. These DC showed low levels of infection similar to peripheral blood DC but in contrast stimulated normal allogeneic T cell proliferation. The capacity of DC exposed to HIV-1 to stimulate T cell proliferation or to show a blocked stimulatory capacity may thus depend not only on the length of the exposure to virus but also on the maturational state of the DC. Loss in DC numbers and function on exposure to HIV 1 may result in lower levels of stimulation of T cells, which in turn may be instrumental in reduction of T cell numbers. PMID- 9225998 TI - Cellular and viral protein interactions regulating I kappa B alpha activity during human retrovirus infection. AB - NF-kappa B/Rel transcription factors participate in the activation of numerous genes involved in immune regulation/inflammation including cytokines, cell surface receptors, adhesion molecules, and acute phase proteins. NF-kappa B activity is controlled by inhibitory proteins, I kappa Bs, that maintain the DNA binding forms of NF-kappa B in an inactive state in the cytoplasm. Many viruses, including the human retroviruses HIV-1 and HTLV-1, also utilize the NF-kappa B/I kappa B pathway to their transcriptional advantage during viral infection. Our recent studies have focused on the I kappa B alpha inhibitor and have characterized several protein interactions that modulate the functional activity of I kappa B alpha during human retrovirus infection. In this article, we summarise recent studies demonstrating that (1) chronic HIV-1 infection of human myelomonoblastic PLB-985 cells leads to constitutive NF-kappa B activity, activated in part due to enhanced I kappa B alpha turnover and increased NF-kappa B/Rel production; (2) HTLV-1 Tax protein physically associates with the I kappa B alpha protein in vivo and in vitro and also mediates a 20- to 40-fold stimulation of NF-kappa B DNA binding activity mediated via an enhancement of NF-kappa B dimer formation; (3) casein kinase II phosphorylates I kappa B alpha at multiple sites in the C-terminal PEST domains and regulates I kappa B alpha function; (4) transdominant forms of I kappa B alpha, mutated in critical Ser or Thr residues required for inducer-mediated (S32A,S36A) and/or constitutive phosphorylation block HIV LTR trans-activation and also effectively inhibit HIV-1 multiplication in a single cycle infection model; and (5) the amino-terminal 55aa of I kappa B alpha (NIK) interacts with the human homologue of dynein light chain 1, a small 9 kDa human homologue of the dynein light chain protein involved in microtubule and cytoskeletal dynamics. Together, our results highlight a number of intriguing molecular interactions between I kappa B alpha and cellular or viral proteins that modulate transcription factor activity and nuclear-cytoplasmic flow of host proteins. PMID- 9226000 TI - Development of laboratory and animal model systems for HIV-1 encephalitis and its associated dementia. AB - The neuropathogenesis of HIV-1 encephalitis and its associated dementia revolves around sustained viral replication in cells of mononuclear phagocyte origin (brain macrophages, multinucleated giant cells, and microglia). Macrophage secretory factors play important roles in facilitating monocyte trafficking into the brain, in regulating productive viral replication, and in producing neurotoxic responses. To study these events, we constructed an artificial blood brain barrier (BBB) to assay monocyte transendothelial migration and developed an animal model system for HIV-1 encephalitis to ascertain the role that virus infected mononuclear phagocytes play in disease pathogenesis. The BBB model was composed of brain microvascular endothelial cells and astrocytes placed on opposite sides of a porous membrane. Monocyte activation, not HIV-1 infection per se, was the central event affecting monocyte BBB migration. Many of the pathological features of HIV-1 encephalitis were reproduced in SCID mice stereotactically inoculated with virus-infected monocytes. These included widespread astrogliosis, apoptosis of neurons, dendritic damage, and macrophage/microglial activation. Such laboratory and animal model systems are being used to ascertain the pathogenic potential of virus-infected macrophages in brain and ways to curb such injurious effects. PMID- 9225999 TI - HIV-1 viral protein R (Vpr) regulates viral replication and cellular proliferation in T cells and monocytoid cells in vitro. AB - Among the putative accessory genes of HIV-1, the 96-amino-acid virion-associated vpr gene product has been described to have several novel biological activities. These include cytoplasmic-to-nuclear translocation, which empowers HIV to infect and replicate in non-dividing cells and to increase viral replication, particularly in macrophages. Along with these viral effects, we found that HIV-1 Vpr induces dramatic biological changes in the target cells of HIV infection, including induction of changes in transcriptional patterns, morphological changes, and complete inhibition of proliferation, which collectively was termed differentiation. These changes occur in the absence of other viral gene products, suggesting that Vpr mediates its proviral effects partially or perhaps solely through modulation of the state of the target cell rather than directly on the virus. The inhibition of proliferation in T cell lines has been extended by several groups to demonstrate that the inhibition of proliferation is through G2 cell cycle arrest, further supporting the idea that Vpr acts directly on cellular targets. We have recently described a role for Vpr in modulating the glucocorticoid pathway, which is involved in the regulation of the state of the cell, in cytoplasmic-to-nuclear translocation, and in modulation of host cell transcription. It is important to note that certain anti-glucocorticoid compounds modulate Vpr activity in vitro. These results support the idea that the host cell contains specific receptor molecule(s) through which Vpr mediates its activity. Consequently, Vpr represents a unique target for anti-HIV drug development and has significance for HIV-1 disease progression. PMID- 9226001 TI - Cellular aspects of HIV-1 infection of macrophages leading to neuronal dysfunction in in vitro models for HIV-1 encephalitis. AB - HIV-1 is a hematogenously spread virus that most likely gains entry into the brain within blood-derived macrophages. Indeed, productive viral replication selectively occurs within perivascular and parenchymal blood-derived macrophages and microglia and HIV-infected macrophages have increased potential to bind and transmigrate through the blood-brain barrier. Once inside the brain, HIV-infected macrophages secrete a variety of pro-inflammatory mediators that display neuromodulatory and neurotoxic activities in several in vitro models for HIV-1 encephalitis. The final outcome regarding neuronal function and cell loss is regulated through intercellular interactions between these virus-infected cells and astrocytes. In this regard, both HIV-induced intracellular events in macrophages and interactions between HIV-infected macrophages and brain cells are reviewed as factors that might lead to neuronal injury in in vitro model systems for HIV-1 encephalitis. PMID- 9226003 TI - IL-13 acts on macrophages to block the completion of reverse transcription, inhibit virus production, and reduce virus infectivity. AB - An understanding of the immune suppression of HIV-1 replication in macrophages continues to be a major goal of AIDS research due to the central role this cell type has in AIDS pathogenesis. We have previously discussed the potential clinical benefits of the anti-inflammatory cytokine interleukin-13 (IL-13), which, unlike IL-4 or IL-10, had limited effects on T cell functions. In this report are extend our observations on the effects of IL-13 on HIV-1 replication in monocyte-derived macrophages (MDM) and show redundancy with IL-4, IL-13 or IL 4 have similar effects on HIV-1 replication in MDM when added at different times after infection, with the ability to decrease infection virus release when added for up to 7 days after infection. Removal of IL-13 from MDM revealed a reduction of infection by 16- to 81-fold based on the absence of viral re-emergence from lower multiplicity of infection (m.o.i.). The reduction of HIV-1 infectivity in MDM caused by IL-13 was further characterized by studies on the formation of viral DNA over a range of m.o.i. IL-13 increased the formation of LTR DNA at the lowest m.o.i. of 0.007 while concurrently inhibiting the formation of gag DNA, a later reverse transcription product, at the highest m.o.i. tested, 0.62. Overall, our data indicate that IL-13 can act on macrophages before and after HIV-1 infection by blocking the completion of reverse transcription, decreasing virus production, and reducing the infectivity of the progeny virions. PMID- 9226002 TI - HIV infection of macrophages and pathogenesis of AIDS dementia complex: interaction of the host cell and viral genotype. AB - AIDS dementia complex (ADC) develops in only a third of HIV-infected patients who progress to AIDS. Macrophages and microglial cells are the major cellular sites of productive HIV replication in brain. Using 11 blood isolates of HIV from asymptomatic patients there was marked variation in tropism and the level of productive infection in recently adherent monocytes and monocyte-derived macrophages cultured in vitro. However, less variation was seen with 19 blood isolates from advanced HIV infection and 11 postmortem tissue isolates from brain, cerebrospinal fluid, spleen, and lung. Newly adherent monocytes expressed CCR5 in all seven patients tested, consistent with their susceptibility to infection but not explaining the above variability. There is, also marked regional variability in neuropathology in the brain of patients with ADC. We have demonstrated that there was marked variation in the V3 sequences of HIV clones from different regions of the cortex of a patient with ADC, suggesting independent evolution of HIV replication in brain. Furthermore, production of the neurotoxin quinolinic acid from HIV-infected macrophages varied, depending on the host and source of HIV isolate. Hence variations in viral genotype, production by infected macrophages, and subsequent toxin production may contribute to the variability in neuropathology between individuals and between different regions of the brain in the same individual. PMID- 9226004 TI - Targeting antiviral nucleotide analogues to macrophages. AB - Macrophages are important target cells for human immunodeficiency virus type 1 (HIV-1) infection. We have developed a drug targeting system for the selective delivery of phosphorylated nucleoside analogues to these phagocytosing cells. This system is based on the possibility of encapsulating the phosphorylated drugs into autologous erythrocytes and on the subsequent selective modification of their membranes to promote macrophage recognition and phagocytosis. Targeted delivery of phosphorylated nucleoside analogues to human, feline, and murine macrophages inhibits the infectivity of HIV-1, feline immunodeficiency virus, and LP-BM5 viruses more efficiently than the administration of the corresponding nucleoside analogues. In vivo administration of 2',3'-dideoxycytidine 5' triphosphate (ddCTP) encapsulated into autologous erythrocytes to LP-BM5-infected mice was found to reduce infectivity and disease progression. Furthermore, the simultaneous administration of AZT or ddC produced additive antiviral effects. The possibility of using red cells as drug targeting systems was useful for the design, synthesis, and delivery of new antiviral nucleoside analogues. As a prototype of these new drugs, di-(thymidine-3'-azido-2',3'-dideoxy-D-riboside)-5' 5'-p1-p2-pyrophospha te (AZTp2AZT) was prepared. Although this drug in solution has the same antiviral activity as AZT, when administered encapsulated into erythrocytes it was several times more efficient in inhibiting the infectivity of human, feline, and murine immunodeficiency viruses. Thus, the availability of a drug targeting system for the selective delivery of antivirals to macrophages offers an additional possibility for the development of new drugs and of new combination antiviral therapies. PMID- 9226005 TI - Inhibition of replication of HIV in primary monocyte/macrophages by different antiviral drugs and comparative efficacy in lymphocytes. AB - Several anti-HIV drugs acting on different steps of virus replication were tested in our experimental model of primary monocyte/macrophages; the results were compared with the activity found in lymphocytes. Nucleoside analogues (AZT, ddI, ddC, d4T, PMEA, 3TC etc.) show greater activity in macrophages (M/M) than in lymphocytes. In particular, the EC50 of AZT, ddC, and ddI in M/M is 2- to 100 fold lower than that found in lymphocytes. This greater efficacy of nucleoside analogues in M/M depends on the enhancement of their chain-terminating activity by the low levels of endogenous deoxynucleoside-triphosphates (dNTP) usually found in resting cells such as M/M. Non-nucleoside reverse transcriptase inhibitors (NNRTI) do not act as chain terminators (thus their antiviral effect is not related to the intracellular concentrations of dNTP); as a consequence the activity of TSAO, HEPT, TIBO, and other NNRTI tested in M/M is similar to that found in lymphocytes. Regarding inhibitors of binding and fusion of HIV, we found that their anti-HIV activity is markedly decreased (or even nullified) when M/M are treated with cytokine activators of M/M function and enhancers of HIV replication. More relevant from a clinical standpoint, protease inhibitors are able to inhibit HIV replication in chronically infected macrophages (i.e., cells carrying the proviral genome already integrated in the host genome). All other inhibitors of late stage of virus life cycle tested (antisense-rev, anti-tat, interferon-alpha and -gamma, phosphorothioate analogues, GLQ-223, etc.) were totally inactive in chronically infected macrophages. The different effects of various classes of HIV inhibitors in lymphocytes and macrophages suggests that AIDS therapy should consider all aspects of the pathogenesis of HIV infection and must be restricted to drugs, or combinations of drugs, active against both lymphocytes and M/M in all body compartments where the virus hides and replicates. PMID- 9226006 TI - Radiopharmaceuticals for breast cancer imaging. PMID- 9226007 TI - Scintimammography (SMM) with 99mTc-MDP: an overview of the experience at the National Cancer Institute of Napoli. PMID- 9226008 TI - 99mTc-sestaMIBI breast scintigraphy. PMID- 9226009 TI - 99mTc-tetrofosmin in breast cancer: experience of the Second University of Naples. PMID- 9226010 TI - The role of internal mammary lymphoscintigraphy in the clinical staging and prognosis of patients with breast cancer. PMID- 9226011 TI - The role of lymphoscintigraphy by periareolar injection in the evaluation of axillary lymph node metastases from breast cancer. PMID- 9226012 TI - Internal mammary chain lymphoscintigraphy (IML) and IML-guided internal mammary chain biopsy (GIMB) in breast cancer. PMID- 9226013 TI - Axillary node metastasis detection in breast cancer with 99mTc-sestaMIBI and 111In-pentetreotide. PMID- 9226014 TI - Optimization of axillary lymphoscintigraphy to detect the sentinel node in breast cancer. PMID- 9226016 TI - The need of cost-effectiveness evaluation when using high-cost equipment in national health services. PMID- 9226015 TI - The contribution of positron emission tomography (PET) with 18F fluorodeoxyglucose (FDG) in the preoperative detection of axillary metastases of breast cancer: the experience of the National Cancer Institute of Milan. PMID- 9226017 TI - Role of bone scan in breast cancer follow-up. PMID- 9226018 TI - Mucinous markers in breast cancer. PMID- 9226019 TI - Radioimmunodetection and radioimmunotherapy of breast cancer. PMID- 9226020 TI - Experience with 89-strontium treatment of painful osseous metastases from breast cancer. PMID- 9226021 TI - Palliative therapy with rhenium-186-HEDP for bone metastases of breast cancer. PMID- 9226022 TI - Rhenium-186 hydroxyethylidene diphosphonate for treatment of painful osseous metastases from mammary carcinoma. PMID- 9226023 TI - Adjuvant hormone therapy after radical prostatectomy: indications and results. AB - Despite recent advances in staging modalities, nearly 30-40% of patients undergoing radical prostatectomy for clinically localized prostate cancer have residual disease. In these cases, one or more of the following conditions may be present: extracapsular disease, positive margins, invasion of the seminal vesicles, lymph node metastases or the postoperative persistence of PSA values above the biological threshold. The optimal management for residual prostate cancer remains controversial and in this setting adjuvant therapy could be appropriate. In the present review we examine the conditions in which hormonal adjuvant therapy can be indicated and the results available from retrospective or non-randomized studies. From the data in the literature and in the absence of randomized prospective studies, prudent conclusions could be drawn on the efficacy of adjuvant hormonal therapy. In cases of small volume, low grade (Gleason score < 7) prostate cancer in stage C or D1, radical surgery coupled with adjuvant hormonal therapy leads to survival rates in stage C similar to those in the intraprostatic stage, and in stage D1 with minimal lymph involvement, seems to delay clinical development of metastases. Finally, the quality of life associated with adjuvant therapy and the drug regimens available for this therapy are reviewed. PMID- 9226024 TI - The potential role of Tomudex in the treatment of advanced colorectal cancer. AB - AIMS AND BACKGROUND: The quinazoline folate analog thymidylate synthase inhibitor, Tomudex, is about to enter the Italian pharmaceutical market. Its place among the therapeutic options for advanced colorectal cancer is discussed. METHODS: The pros and cons of currently available chemotherapeutic regimens are briefly described with special attention to patient's and tumor's determinants of treatment outcome. The mechanism of action and the results of phase I, II and III studies of Tomudex are reviewed. RESULTS: Not all patients need to be treated. Guidelines are given in this respect. Tomudex at the dose of 3 mg/m2 given i.v. every three weeks has antitumor activity similar to that of currently available regimens, with a favorable toxicity profile. CONCLUSIONS: Current research approaches are unlikely to dramatically improve the treatment outcome of this disease in the near future. What can reasonably be expected is less toxicity and more convenient routes and schedules of drug administration that may translate into better quality of life for our patients. Tomudex has been devised along these lines. PMID- 9226025 TI - Quality of life among mastectomy patients using external breast prostheses. AB - BACKGROUND: Most women who undergo mastectomy for breast cancer use external breast prostheses. Yet, little is known about patterns of use, satisfaction levels, and quality of life associated with their use as compared to other options. PATIENTS AND METHODS: We report longitudinal, self-report questionnaire data regarding prosthetic use from 592 Italian mastectomy patients. Women who report satisfaction with their prostheses are compared on medical, demographic, and quality of life variables to a matched sample of women who report dissatisfaction. We also compare matched samples of women who do not use prostheses and women who had reconstruction to prosthetic users. RESULTS: Most women used and were satisfied with their prostheses. However, there was a small group of women who were dissatisfied. These women reported greater disruption to their sense of feminility and worse quality of life in some areas. We found few differences between prosthetic users and women who used either of the other two options available following mastectomy-taking no action to restore the appearance of the amputated breast or having reconstructive surgery. CONCLUSIONS: No one technique for restoring the appearance of the mastectomized breast is necessary to optimize quality of life for all women. Physicians should describe the options to women, along with the average satisfaction rates for women choosing those options, and help women to make the best personal decisions. PMID- 9226026 TI - Combined and sequential expression of p53, Rb, Ras and Bcl-2 in bronchial preneoplastic lesions. AB - AIMS AND BACKGROUND: Several simple molecular abnormalities have been detected in bronchial preneoplastic lesions, but the simultaneous presence of these alterations has been scarcely investigated. METHODS: We studied, by an immunohistochemical method, the expression of p53, Rb, Ras and Bcl-2 in 65 samples from surgical specimens and diagnostic biopsies selected for the presence of preneoplastic changes in the bronchial epithelium. To perform an analysis of the combined expression of all markers in the same areas, we accurately mapped every consecutive section on which immunohistochemical reactions were performed, subdividing each specimen into 25x microscopic fields, which allowed good topographical mapping. RESULTS: It was found that the frequency of p53-positive and Rb-negative microscopic fields was directly related to the morphological grading of lesions. On the other hand, Ras expression characterized high-grade lesions not showing squamous differentiation (non-squamous Cis). Regarding Bcl-2 expression, only slight differences in positivity distribution were found between the different lesions. More interesting was the parallel evaluation of all markers in the same areas: one of the main patterns, found to be correlated with the severity of histopathological features, was characterized by combined p53 hyperexpression/Rb hypoexpression; furthermore, when Ras and Bcl-2 hyperexpression were superimposed to the above pattern, the former mainly characterized non-squamous Cis, while the latter was present only in high-grade squamous lesions. However, the most frequently encountered pattern did not show any alteration of the studied markers, suggesting that other mechanisms could be involved in bronchial carcinogenesis. CONCLUSIONS: The detection of combined molecular abnormalities in bronchial preneoplasia could clarify the steps involved in lung carcinogenesis; furthermore, a simple and inexpensive method, such as immunohistochemistry, could be routinely applied also to cytologic specimens in order to detect those lesions, or patients, that are prone to progression towards lung cancer. PMID- 9226027 TI - OVCA (CA125) second generation: technical aspects and serum levels in controls, patients with liver disease, pregnant women and patients with ovarian disease. AB - An immunoradiometric method of the second generation (IR-MA II) is widely used to determine CA125 serum levels. In this study we have evaluated the performance characteristics of a commercially available IRMA CA125 II (Byk-Gulden, Sangtec Diagnostica). The CA125 serum levels were determined in several groups of patients (healthy women, pregnant women, subjects affected by benign and malignant ovarian cancer, patients with liver diseases) with two IRMAs CA125 II (Byk-Gulden, Sangtec Diagnostica and Centocor, Diagnostic Division) and IRMA CA125 I (Byk-Gulden, Sangtec Diagnostica). Our results show a good analytic performance of IRMA CA125 II (Byk-Gulden, Sangtec Diagnostica), a good correlation between IRMAs CA125 II (Byk-Gulden, Sangtec Diagnostica and Centocor, Diagnostic Division), but an unacceptable correlation between IRMAs CA125 II (Byk Gulden, Sangtec Diagnostica and Centocor, Diagnostic Division) and IRMA CA125 I. A statistically significant difference was observed comparing the values obtained with both IRMAs CA125 II and IRMA CA125 I in the groups of patients. In contrast no statistically significant difference was observed when we compared the values obtained with IRMA CA125 II (Byk-Gulden, Sangtec Diagnostica) and IRMA CA125 II (Centocor, Diagnostic Division). CA125 serum values obtained with the second generation kits were different from those obtained with the first-generation one; consequently, it is important, especially in the follow-up of cancer patients, that CA125 serum values be obtained with kits of the same generation. Our data seem to suggest the use of second-generation kits to determine CA125 serum levels. PMID- 9226028 TI - Postoperative adjuvant chemoradiotherapy for rectal cancer: analysis of acute and chronic toxicity. AB - AIMS AND BACKGROUND: The aim of the study was to evaluate acute and chronic toxicity of combined postoperative standard radiation therapy to the pelvis and 5 fluorouracil plus levamisole in resectable rectal cancer. METHODS: Between July 1990 and September 1993, 58 patients with histologically confirmed adenocarcinoma of the rectum entered the prospective study. The schedule consisted of 5 fluorouracil, 450 mg/m2 i.v. for 5 days, and from day 28 5-fluorouracil, 450 mg/m2 i.v. weekly for 24 weeks, plus levamisole given orally at the dose of 150 mg every day for 3 days every 2 weeks for 6 months; radiotherapy (180 cGy/day) 5 days a week for a total dose of 45 Gy was administered from day 28. RESULTS: After the first cycle of chemotherapy (before radiotherapy), overall toxicity was mild. During chemoradiotherapy, dose-limiting toxicity was grade 3 diarrhea and proctitis, for which the combined treatment was interrupted for more than 7 cumulative days in 28 patients. During the 24 weeks of weekly 5-fluorouracil (after radiotherapy), no severe toxicity was reported. Three-year survival and progression-free survival were 65% and 50-55%, respectively. CONCLUSIONS: Although adjuvant chemoradiotherapy is usually feasible, in our study toxicity was severe in a substantial proportion of patients, probably due to the schedule applied. We are evaluating the feasibility and toxicity of a combined treatment which includes 5-fluorouracil in continuous chronomodulated infusion during radiotherapy. PMID- 9226029 TI - Ifosfamide and etoposide in previously treated patients with advanced breast cancer. AB - AIMS AND BACKGROUND: Ifosfamide is an active alkylating agent in the treatment of breast cancer, as a first-line therapy and in advanced disease. Since the combination of etoposide with an alkylating agent produces a synergistic and tolerable activity in various malignancies, in the present study, ifosfamide and etoposide were administered to patients with advanced breast cancer to evaluate the response characteristics and the toxicity profile. STUDY DESIGN: The combination of ifosfamide, mesna and etoposide was prospectively administered to 41 previously treated patients with stage IV breast carcinoma. The treatment schedule consisted of ifosfamide, 1500 mg/m2, infused over 24 hrs with 1500 mg/m2 mesna on days 1 to 5 and 120 mg/m2 etoposide, infused over 1 hr on days 1 to 3, to be repeated every 4th week. RESULTS: After a median follow-up of 10 months, an objective response rate of 23% (overall 2.5% complete remission and 20.5% partial remission) and a median response duration of 5.3 months were obtained in 39 assessable patients. The non-responder group consisted of 28.3% stable disease and 48.7% progressive disease. The prior status of chemotherapy was the only significant prognostic factor with an impact on the response rate. The overall toxicity was generally mild, with grade 3 myelotoxicity encountered in 25.7% of patients. CONCLUSIONS: The tolerable side effect profile of the ifosfamide and etoposide combination might be advantageous as regards the quality of life. To improve the rate and/or the duration of response and to clarify the precise role of the ifosfamide-etoposide combination in previously treated advanced breast cancer, further trials are warranted. PMID- 9226030 TI - Brain metastases in well-differentiated carcinoma of the thyroid. AB - Fifteen patients (4 males and 11 females) developed brain metastases from well differentiated thyroid cancer within 1 month to 14 years of the initial diagnosis. One patient presented with a brain tumor. Except for 3 patients with unique brain metastases, all the others had extensive metastases in nodes, lungs and bones in various combinations. Brain metastases generally appeared after the onset of metastases at other sites. The histology of the brain tumor matched the primary pathology in the 6 operated cases. The treatment was surgery and external radiation in 6 cases, and radioiodine or chemotherapy in the others. Survival in general was less than 6 months after the diagnosis of brain metastases. The prognosis is poor once the onset of brain metastases is evident. PMID- 9226031 TI - Unusual breast lumps. AB - Two new Italian cases of breast infection by Dirofilaria Repens are presented. In one case the correct diagnosis was clinically achieved through needle aspiration. This case is documented by a surprising macrophotography. PMID- 9226032 TI - Clinical determinants of long-term survival in patients with glioblastoma multiforme. AB - Repeated reports of more than ten years postoperative survival in patients with glioblastoma multiforme (GM) have appeared in the literature over the last decades. Authors have tried to identify the clinical, therapeutic and histological features determining long-term survival. We present two patients in whom, after radical removal of the tumor followed by conventional radiation, there has been no recurrence for at least ten years. The young age of the patients and the radical surgical approach were in accordance with previous reports of long-term survival. Nevertheless, one tumor originated from the thalamus, a location considered to be of unfavorable prognosis. We therefore further discuss the value of clinical signs as determinants in the prognosis of GM. PMID- 9226034 TI - Disseminated nocardiosis in a metastatic breast cancer patient. PMID- 9226033 TI - Tuberculous skeletal muscle involvement in acute leukemia: report on two cases. AB - Bacterial infection of skeletal muscle (pyomyositis) is usually followed by abscess formation. The most commonly isolated pathogen is Staphylococcus aureus. Tuberculosis rarely affects patients with acute leukemia. The authors report on 2 patients, one with acute myelogenous leukemia and the other with acute lymphoblastic leukemia whose clinical course was complicated by tuberculous skeletal muscle abscesses. In both instances, musculoskeletal pain was accompanied by evidence of muscle abscesses by imaging studies of the painful areas. Therefore, in patients with acute leukemia and evidence of muscle abscesses with initial cultures negative for bacteria and fungi, one should include tuberculosis in the differential diagnosis. PMID- 9226035 TI - Palliative care and euthanasia, still confusion? PMID- 9226037 TI - Changing patterns of cell adhesion molecules during mouse pelage hair follicle development. 2. Follicle morphogenesis in the hair mutants, Tabby and downy. AB - Wild-type mice have three main types of hair in their pelage: tylotrichs, awls and zigzags. Tabby mice have a yellowish coat consisting of awls only, whereas downy mice have a sparse grayish coat consisting of unusually fine hairs. The spatial and temporal distribution of cell adhesion molecules (CAMs) during hair follicle morphogenesis was investigated in the mutants and compared with that in nonmutant mice. In Tabby embryos, awl follicles developed normally and showed normal immunostaining patterns for E-cadherin, P-cadherin and N-CAM. Prior to follicle initiation, however, some deviations from normal skin morphology and staining patterns indicated a delay in the development of the basal epidermal layer. On the other hand, the stratum corneum was formed prematurely. Therefore, the lack of tylotrich and zigzag follicles in Tabby mice might be explained by a general defect in epidermal development rather than by abnormal CAM expression. In downy embryos, tylotrich and awl follicles were initiated within the normal time periods, but elongation and differentiation of most follicles were abnormal. At birth, most follicles were small and/or severely deformed but showed normal CAM expression patterns. Extreme distortion and disorientation of follicles seemed to be associated with disintegration of the dermal papilla and abnormal mesenchymal cell condensations between the follicles. This suggests that abnormal hair development in downy mice might result from a defect in dermal rather than epidermal components of the skin. PMID- 9226036 TI - Changing patterns of cell adhesion molecules during mouse pelage hair follicle development. 1. Follicle morphogenesis in wild-type mice. AB - The morphogenesis of hairs is initiated and maintained by reciprocal interactions between groups of epithelial and mesenchymal cells. To examine whether cell adhesion molecules play a role in this process, prenatal distribution patterns of various cell adhesion molecules were studied during hair follicle morphogenesis in the dorsal skin of C57BL mouse embryos, using monoclonal antibodies. E cadherin was present on all epithelial cells of the skin when the ectoderm gave rise to periderm and epidermis. E-cadherin was reduced in the follicle placodes and hair plugs, then disappeared from the presumptive hair matrix of elongating follicles. P-cadherin was initially present on all cells of periderm and epidermis and was later retained at a reduced level in the basal epidermal layer. P-cadherin was prominent in all follicle placodes and hair plugs and in the presumptive hair matrix of elongating follicles. N-CAM was present on all mesenchymal cells of the presumptive dermis at the prefollicle stage, then temporarily restricted to a few cells just below the dermal-epithelial junction. Later, N-CAM reappeared in the interfollicular mesenchyme and was prominent in the mesenchymal sheath and dermal papilla of elongating follicles. In addition, N CAM was expressed in the hair plugs, then became progressively restricted to the upper caudal part of the elongating follicles. The results suggest that the main role of cell adhesion molecules is to mould the follicle by relaxing or reinforcing cell contacts in areas of increased morphogenetic activity. PMID- 9226038 TI - Apoptosis in adult mouse testis induced by experimental cryptorchidism. AB - Induction of cryptorchidism in the mouse causes infertility due to disruption of spermatogenesis including reduction of germ cells; however, the cellular mechanism responsible for the degenerative changes in cryptorchid testis is still unclear. In surgically induced bilateral cryptorchidism of 3-month-old C57BL/Tw mice, cellular changes in the cryptorchid testis were studied 1, 2, 3, 7, 14 and 21 days after the operation by electron microscopy, DNA fragmentation, in situ 3' end labeling, serum and testicular testosterone measurements and gene expression. Although the testis showed DNA fragmentation even in intact mice, the cryptorchidism increased the degree of the fragmentation at 1 postcryptorchidism (p.c.) day. Apoptosis was encountered mainly in spermatids and spermatocytes. The number of apoptotic cells in the cryptorchid testis showed a 7-fold increase at 1 p.c day as compared to the intact testis, then it gradually decreased. Serum testosterone levels showed a significant decrease at 2 p.c. days and remained low thereafter. Expression of transforming growth factor-beta 2 (TGF-beta 2), TGF beta 3, tumor necrosis factor-alpha receptor and Fas mRNAs increased in the cryptorchid testis within 24 h after the operation. In lpr(cg) and lpr mice lacking functional Fas, gld mice lacking functional Fas ligand and lpr(cg)-gld mice lacking both functional Fas and Fas ligand, the experimental cryptorchidism also induced apoptosis in germ cells at 1 p.c. day. The present results indicate that cryptorchidism induces apoptotic dell death in germ cells, and that testosterone reduction and the Fas system may not be significantly involved in the apoptosis of male germ cells. PMID- 9226039 TI - Functional sexual differences in rat mammillary bodies: a quantitative Ag-NOR study. AB - The mammillary bodies of the posterior hypothalamus are one of the CNS structures in which structural sexual dimorphism has already been described. This study quantifies the argyrophilic nucleolar organizer regions (NORs) of the neurons from this region in the male and female rats, the latter during two major phases of the estrous cycle (estrus-diestrus). The number and relative area of these NORs stained with a silver nitrate technique are considered as an index of the global protein synthesis of the neurons in the different nuclei from the mammillary bodies. Our results show the existence of statistically significant differences between sexes and estrous cycle phases. These findings suggest a significant influence of gender and the hormonal state on the neural activity of the MB. PMID- 9226040 TI - Cytoarchitecture of the abducens nucleus of man: a Nissl and Golgi study. AB - The abducens nucleus is a pontine nucleus directly involved in oculomotion through its connections with the lateral rectus muscle of the eye. The aim of the present study was to investigate the cytoarchitectural organization of the abducens nucleus in man. The data obtained showed that the nerve cell bodies were small, medium and large in size and polygonal, oval, round or spindle shaped. The cytoplasm of all neurons appeared basophilic due to clearly evident scattered Nissl granules. On the basis of the characteristics of the dendritic arborization, multipolar and fusiform cells were identified. The multipolar neurons showed four to eight primary dendrites which gave off a wide secondary ramification. The fusiform neurons showed two dendrites emerging from the opposite poles of the elongated nerve cell body. The dendrites of all the neurons were largely confined within the boundaries of the nucleus. This finding would suggest that the neuronal relationships of the abducens nucleus supplied by the afferent fibers which pass through or end within it take place almost completely inside the nucleus. The wider dendritic arborization shown by the multipolar cells would indicate the latter as the principal target fields for the afferent inputs. PMID- 9226041 TI - Extraocular motoneuron stimulation frequency effects on motor unit tension in cat. AB - The contractile characteristics of 47 twitch and 3 nontwitch lateral rectus motor units in 11 cats were examined using two different stimulation paradigms derived from observed motoneuron firing frequencies in alert animals. The twitch units showed an average twitch tension (46.0 +/- 8.1 mg), contraction time (6.15 +/- 0.26 ms) and median fusion frequency (170 Hz) consistent with previous studies, kt value, defined as the slope of the relation between a series of constant frequency tetanic stimulation trains (lasting 200 ms) and tetanic tension, correlated with maximum tetanic tension (r = 0.984, p < 0.05). ktps value, defined as the slope of the relation between tetanic tension and a series of constant frequency stimulation trains (lasting 1975 ms) that immediately followed a 25-ms high-frequency burst (pulse/step paradigm), was similarly correlated (r = 0.853, p < 0.05) with maximum tetanic tension (x = 256.5 +/- 35.2 mg). ktps values were lower that kt values for each unit, but the units did not change their position in the numerical hierarchy. Eighty-four percent of the motor units produced different maximum tetanic tensions (x = 24 +/- 3%), when comparing pulse/step to constant frequency stimulation, but only 14% of those units produced a greater maximum tetanic tension during pulse/step stimulation. In contrast, 46% of the motor units contracted with more force during the step phase of the pulse/step paradigm than with constant frequency stimulation when the stimulation rate was below 120 Hz: 24% of the units contracted with less force. In addition, pulse/step stimulation frequency ranges above 120 Hz (step phase) were ineffective in increasing tension while higher frequencies continued to elicit tension increases during constant frequency stimulation. The impact of these tension variations on eye movement is discussed. PMID- 9226042 TI - Variability in the human entorhinal region may confound neuropsychiatric diagnoses. AB - The human entorhinal region consists of a number of areas; however, there is no generally accepted nomenclature for these cytoarchitectonic fields, and the designation of its constituent layers or strata is a matter of controversy. Here, we consider a hitherto neglected adjacent field, the preamygdaloid claustrocortex. Its medial subfield has a small common border with the rostromedial entorhinal region (width maximal 2 mm). Both fields are cytoarchitectonically rather similar. The rostromedial oral entorhinal field lacks ascending terminal islands. Its unusually small pre-alpha cells are arranged in a thin band or small clusters consisting of pyramidal, triangular, or polymorphic cells. The conspicuous chromophilic pre-beta cell clusters are composed of a variety of cell types, including groups of 'immature' spindle shaped or bipolar nerve cells. Furthermore, a rare sulcus within the entorhinal region (central sulcus of the entorhinal region: observed in 4% of the 450 brains examined) is associated with an unusual lamination of the entorhinal layers in its wall and floor. Both the specific shape and arrangement of neurones in the claustrocortical-rostral entorhinal border region and the unusual lamination within the rare central entorhinal sulcus are regarded as reflecting neurodevelopmental disturbances characteristic of schizophrenic brains. In contrast, our observations in a large sample of serially sectioned brains from controls, schizophrenics, and patients suffering from neuropsychiatric diseases other than schizophrenia do not support this assumption. PMID- 9226043 TI - Myocardial bridge muscle on left anterior descending coronary artery differs from subepicardial myocardium of the left ventricle in dogs. AB - The myocardial bridge (MB) is a muscle band found sporadically above the coronary artery (CA) in humans and certain animals such as the dog, cat and sheep. The purpose of our study was to compare the structure of the MB muscle with that of tissue from the subepicardial myocardium. The histological studies included toluidine blue staining of 1-micron-thick sections and Gomori's trichrome staining of canine cardiac samples. The MB muscle of the dog heart is characterized by a distinctive spatial arrangement, with individual fibers separated by substantial elements of intercellular connective tissue in cross section. Longitudinally, the long, slender fibers are aligned continuously with intermediation of intercalated disks lying perpendicular to the long axis of the fibers. In other regions of the left ventricular subepicardial myocardium, each myocyte is tightly packed in transverse view. There is great variation in the thickness (0.11-2.24 mm, average 0.45 mm) of MBs and the distance (24-236 microns, average 103 microns) between MBs and the left anterior descending coronary artery (LAD) among the 13 affected dogs examined, with no apparent relationship between the occurrence of MBs and either age or sex. These results on MB alignment suggest that the MB muscle generates force along the long axis of the fiber orientation as skeletal muscle does, and with minimal constriction of the CA; if so, the function of MB myocytes may differ from that of common cardiac myocytes, as does the structure. Then, the long-supposed downward compressive force of MBs on the LAD would be minimal in most cases; however, when the MB produces a systolic narrowing of the LAD known as the milking effect, the degree of lateral compression and its influence should depend not only on the substantial size of the MB muscle but also on the distance between the MB muscle and LAD. The environment surrounding the LAD may be a crucial factor in determining whether the MB influences the induction of heart disorders or not. PMID- 9226044 TI - Myocardial bridging as a factor in heart disorders: critical review and hypothesis. AB - The purpose of this report is to review the previous and current methods for the detection of a MB and to summarize results of work by other investigators as well as our own recent morphological studies with emphasis on work since 1983. We will discuss the presumable association of MBs with heart disorders, on the basis of significant publications. We describe the importance of both basic and clinical research for the assessment of MB influence in various heart disorders. Aspects covered are: (1) historical background of MBs and the relationship between the MB and CA; (2) general characteristics of MBs; (3) methods for studying MBs; (4) compressive effect of the MB on the CA; (5) clinical symptoms including myocardial ischemia, angina pectoris, myocardial infarction, development of atherosclerosis and thrombosis, ventricular fibrillation and sudden death; (6) treatment of MB-associated heart disorders by resection of MBs and (7) future research trends. PMID- 9226046 TI - Auditory brainstem response: recent developments in recording and analysis. AB - During the past 25 years since the 'discovery' of the ABR, clinicians world-wide have exploited the clinical utility of this evoked response with many different populations. The 1970s and 1980s saw the development of ABR protocols for increasing the reliability and validity of the response in various clinical situations. Now, in the last decade of the century, advanced technology and the development of increasingly sophisticated software allows more flexibility in these measurements. Thus, ABR testing continues to evolve at a rapid pace. The main focus for ABR in the 1990s has been on developing techniques to improve the quality of the response, decrease the time in which it takes to record the response, and minimize human error through objective response analysis. Some of the major recent developments were reviewed in this chapter. The intent of this review is to provide the researcher and the clinician with an understanding of the underlying principles of the techniques which will be, and are already, available to them on their ABR equipment. Many of these techniques are promising for use with newborns, children, and difficult-to-test populations where time is a limiting factor in the amount of information that can be obtained. We encourage any and all clinicians to experiment with these techniques to determine how each technique might contribute to their particular clinic protocols. As with any new technique, there will be a learning curve associated with understanding the underlying principles, mastering the technical requirements, and realizing the most valuable clinical applications of these methods. Once the procedures become familiar to clinicians and applied in varied clinical settings, it is likely that the neurodiagnostic and audiologic value of ABR will be profoundly enhanced. PMID- 9226045 TI - Electrocochleography. PMID- 9226048 TI - Objective measurements and the audiological management of cochlear implant patients. PMID- 9226049 TI - Electroneuronography. PMID- 9226047 TI - Hearing as reflected by middle and long latency event-related potentials. PMID- 9226050 TI - Testing the vestibulo-ocular reflex. PMID- 9226051 TI - Vestibular evoked potentials. PMID- 9226052 TI - Otoacoustic emissions. PMID- 9226053 TI - New routes and formulations for allergen-specific immunotherapy. PMID- 9226054 TI - Mechanism of aspirin-induced asthma. PMID- 9226055 TI - Systemic effects of two nasally administered glucocorticosteroids. AB - Two topical corticosteroids, budesonide (BUD) and beclomethasone dipropionate (BDP), both administered as suspensions in water, were investigated in healthy volunteers regarding influence on cortisol in plasma and urine (U-cortisol) after nasal application. In the first study, single doses of 200, 400, and 800 micrograms of BDP and BUD were given at 10:00 pm. In the second study, 100, 200, and 400 micrograms were given mornings and evenings for 4 days. In the single dose study, none of the drugs or doses showed any significant influence on cortisol in plasma. However, U-cortisol decreased significantly after BUD 400 and 800 micrograms. In the multidose study, U-cortisol values were significantly reduced after all doses of BUD and the highest dose of BDP. The compounds tested showed different ability to cause measurable systemic effects after nasal application. The clinical implication is that the prescriber, when choosing a compound, should take the application site into consideration and should also be encouraged to find the lowest effective dose. PMID- 9226056 TI - The effects of topical nasal steroids on rat respiratory mucosa in vivo, with special reference to benzalkonium chloride. AB - Fifty rats were treated with topical nasal steroids with and without the preservative benzalkonium chloride in their right nostril twice daily for 21 days, while the left nostrils were exposed to 0.9% NaCl. By cutting the noses serially in frontal sections, the structure of the mucosal lining of all parts of the nose could be investigated. Areas with squamous cell metaplasia were observed in all nostrils exposed to topical steroids containing benzalkonium chloride. Such alterations were not observed in any nasal cavities exposed to the topical nasal steroid without the preservative or to 0.9% NaCl. In conclusion, benzalkonium chloride appears to be potentially toxic to the mucosa in vivo. PMID- 9226057 TI - Occupational allergy in saffron workers. AB - Sensitization to the flower of saffron, a plant commonly grown in Spain for commercial purposes, and its clinical significance as an occupational allergen were studied. The prick test and RAST, with saffron pollen, stamen, and pistil extracts, were used to evaluate the cutaneous and specific antibody responses in the studied population. Provocation tests in patients with clinical findings were used to verify the implication of saffron components in these symptoms. Fifty saffron workers were evaluated. Three of them were sensitized to saffron pollen and stamen proteins, giving prick and RAST positive values. One patient presented asthma, showing a positive bronchial provocation test, and two patients rhinoconjunctivitis, showing positive conjunctival provocation tests. Of a general allergic population of 237, 10 patients also presented cutaneous test and IgE positive to saffron. Saffron allergens (from pollen and stamens) were characterized by SDS-PAGE immunoblotting. A relevant allergen of 15.5 kDa with profilinic nature was detected and further purified by high-resolution gel filtration chromatography. No allergenic components were demonstrated in pistils. Cross-reactivity of saffron extracts was evaluated by RAST inhibition with respect to other pollen species commonly causing sensitization in the same area of study. A significant degree of cross-reactivity was demonstrated between saffron and Lolium, Salsola, or Olea. The identification of the protein components involved in the cross-reactions was investigated by blot inhibition. PMID- 9226058 TI - Dust-mite-allergen concentrations in asthmatics' bedrooms in the Quad Cities (Illinois, USA) after the Mississippi River floods of 1993. AB - This study aimed to measure allergens from Dermatophagoides farinae and D. pteronyssinus and to examine possible relationships of these mite allergens with flooding and other housing factors. A total of 313 dust samples were collected from the bedrooms of 57 asthmatics in 45 homes in the Quad Cities (Illinois, USA) and analyzed by ELISA for the presence of the D. farinae and D. pteronyssinus allergens. Twenty of these homes had some flooding in the last 12 months due to the Mississippi River floods of 1993 and/or other factors. The log-transformed least-squares means of allergens collected were 28 ng/m2 for the D. farinae allergen and 26 ng/m2 for the D. pteronyssinus allergen. D. farinae allergen levels were significantly higher in homes located in the valley, in homes during the summer months, in homes with furred or feathered pets, in homes which had not been flooded in the last year, and in homes where rugs had been steam-cleaned in the last 12 months. D. pteronyssinus allergen levels were significantly higher in homes located in the valley, in homes during April and July-September, in homes with furred or feathered pets, and in homes with no dehumidifier. PMID- 9226059 TI - Adjunct effect of loratadine in the treatment of acute sinusitis in patients with allergic rhinitis. AB - H1-blockers are often added to the standard treatment of acute sinusitis, but this is not supported by a controlled study. A multicentric, randomized, double blind, placebo-controlled, parallel-group study was done in 139 allergic patients (15-65 years) to assess the adjunct efficacy of loratadine in acute exacerbation of rhinosinusitis. Sinusitis was diagnosed by symptoms and confirmed by rhinoscopy and sinus radiograph. Allergy was characterized by skin tests, RAST, and history. Patients were treated with antibiotics (14 days), oral corticosteroids (10 days), and loratadine (10 mg OD) or placebo (28 days). Treatment efficacy was assessed over 28 days by symptom scores quoted daily by patients. Physicians also rated total symptom scores at entry and at day 28. At entry, both groups had similar symptoms. Placebo-treated patients improved significantly, but patients who received loratadine had a significantly greater improvement in sneezing (P = 0.003) after 14 days, and in nasal obstruction (P = 0.002) after 28 days. Physicians found that patients receiving loratadine were significantly improved compared to placebo patients (P = 0.0125). Loratadine in addition to standard therapy was found to improve the control of some symptoms of sinusitis. PMID- 9226060 TI - Occupational type I allergy to Christmas cactus (Schlumbergera). AB - The study aimed to determine whether occupational contact urticaria and symptoms of mucous membranes, reported by five workers in a cactus nursery, were due to IgE-mediated allergy to Schlumbergera cacti. The five persons had positive skin prick tests to the plants as is and positive histamine-release tests, and in three of them specific IgE to the cacti could be demonstrated by Maxisorp RAST and immunoblotting. Four of the patients were atopic, and the fifth had a positive skin prick test to cat dander, indicating latent atopy. Skin prick tests with cacti were negative in most atopic volunteers, and all had negative histamine-release tests. The results suggest a true IgE-mediated allergy to the cacti, and both genetic predisposition and close contact with the plants at work seem to be important factors in the emergence of this new occupational allergy. PMID- 9226061 TI - Protein and allergen content of various natural latex articles. AB - Proteins remaining in products made of natural rubber latex are potential sensitizers. In the present work, we quantified the releasable protein and allergen contents in 37 brands of latex gloves and 26 other latex products. Our results demonstrate the presence of widely varied protein and allergen contents in various latex articles and the lack of a correlation between the protein and allergen values. These findings may assist hospital management and medical staff to take effective preventive measures. PMID- 9226062 TI - IgE-mediated hypersensitivity to latex in childhood. AB - A total of 267 children scheduled to receive anesthesia during a surgical, neurosurgical, or orthopedic intervention were investigated. IgE antibodies against latex were detected in serum samples of 6.4% (17/267 children) of the patients. The most important difference between sensitized and nonsensitized children was the number of surgical interventions in the past. The median of surgical interventions was 1.0 in the nonsensitized group of children and 3.0 in the sensitized group. Only 0.9% of the children with up to two surgical interventions and 34.1% with three or more procedures were sensitized to latex. Only one of the sensitized children developed intraoperative anaphylaxis during intervention after our investigation. We conclude that children with a history of three or more surgical interventions have a high risk of sensitization to latex proteins. Nevertheless, the predictive value of IgE antibodies against latex for development of anaphylaxis during anesthesia seems to be low. PMID- 9226063 TI - The prevalence of latex allergy in children seen in a university hospital allergy clinic. PMID- 9226064 TI - IgE response to penicillin. PMID- 9226065 TI - Dietary and environmental preventive measures for high atopic risk babies. PMID- 9226066 TI - Allergy to frogs. PMID- 9226067 TI - Immediate local reaction to tetanus toxoid booster. PMID- 9226068 TI - Immediate reaction to methylprednisolone with tolerance of other corticosteroids. PMID- 9226069 TI - Association of cold urticaria and aquagenic urticaria. PMID- 9226070 TI - Hymenoptera stings in conscripts. PMID- 9226071 TI - Allergy and the premenstrual syndrome (PMS). PMID- 9226072 TI - Prolonged continuous i.v. "desensitization" in Tienam hypersensitivity. PMID- 9226073 TI - Oral allergy syndrome from pork. PMID- 9226074 TI - Allergic conjunctivitis from eyedrops. PMID- 9226075 TI - Allergic contact dermatitis from mitomycin C. PMID- 9226076 TI - Exercise-induced egg anaphylaxis. PMID- 9226077 TI - Allergic disease prevalence in Izmir. PMID- 9226078 TI - Allergy to dental prosthesis. PMID- 9226079 TI - A review of the use of augmentation therapy for the treatment of resistant depression: implications for the clinician. AB - OBJECTIVE: To critically review the literature on augmentation therapy in resistant depression in order to assist the clinician to make a reasoned choice. Augmentation therapy is defined as the addition of a second agent to an existing antidepressant regimen with the aim of achieving improved clinical response. METHOD: The available literature which related specifically to currently popular augmentation strategies in treatment resistant depression for the past 20 years was examined. The scientific evidence supporting the efficacy of these regimens and their safety was reviewed. RESULTS: Considerable research on lithium augmentation has been undertaken, and on triiodothyronine augmentation to a lesser degree. A number of other drugs have been trialed as augmentation agents with claims of success; however, most of the evidence supporting these agents is anecdotal and in the form of case reports. There are very few well-performed double-blind placebo-controlled studies of augmentation therapy. CONCLUSIONS: Because of possible complex pharmacodynamic and pharmacokinetic interactions, augmentation therapy is not without its potential complications. Lithium augmentation of tricyclic antidepressants can be recommended as a safe and effective strategy and there is a body of scientific evidence supporting the addition of T3 as an effective augmentation agent. Recent research with pindolol augmentation of selective serotonin re-uptake inhibitors (SSRIs) is encouraging, but these findings require replication. There is no empirical evidence supporting buspirone, carbamazepine, sodium valproate, methylphenidate or amphetamine as effective augmentation agents, or that adding a tricyclic to a SSRI has usefulness in relieving depressive symptoms. There is a need for considerable research in this area, with more prospective well-controlled placebo studies. PMID- 9226080 TI - The assessment of depression in the postnatal period: a comparison of four self report questionnaires. AB - OBJECTIVE: The objective of this study was to ascertain the degree of agreement between four self-report depression scales, with particular emphasis on whether each scale would identify the same subgroup of women as being 'most depressed'. METHOD: The questionnaires were administered to a sample of approximately 200 postnatal women at 4 weeks, 4 months and 8 months after delivery. The instruments were: the Edinburgh Postnatal Depression Scale; the depression subscale of the Hospital Anxiety Depression Scale; the Zung Self-Rating Depression Scale; and the depression subscale of the Profile of Mood States. RESULTS: Agreement between pairs of instruments, in terms of identifying the most depressed subgroup of women in the cohort, only averaged approximately 40%. Agreement between the three instruments was only about 25%. CONCLUSIONS: This poor level of agreement most likely reflects the different emphasis in item content of the questionnaires, which in turn reflects different notions of 'depression' held by the designers of the instruments. The implications of the findings for research and clinical practice are discussed. PMID- 9226081 TI - Vietnamese and Arabic women's responses to the Diagnostic Interview Schedule (depression) and self-report questionnaires: cause for concern. AB - OBJECTIVE: The original study aimed to determine the best cut-off scores to screen for postnatal depression on translated versions of the Edinburgh Postnatal Depression Scale (EPDS) for Vietnamese and Arabic women. This research was conducted using the depression module of the Diagnostic interview Schedule (DIS) to determine caseness. This paper reports on the suitability of this diagnostic interview as a criterion measure of depression in these women with a non-English speaking background. METHOD: Vietnamese and Arabic women in south-west Sydney completed the EPDS and a General Health Questionnaire (GHQ-30) antenatally. At 6 8 weeks postpartum they completed an EPDS, the GHQ-30 and a Faces Scale, and were interviewed using the depression module of the DIS. Members of a small convenience sample of women were asked about the cultural appropriateness of each of the instruments. RESULTS: Vietnamese women admitted to few depressive symptoms on the DIS, whereas they appeared more open to reporting these on the EPDS and the GHQ-30. Arabic women responded more openly to the questionnaires and the interview, although they too were reluctant to report specific symptoms on the DIS. CONCLUSION: The usefulness of the DIS in determining rates of major depression in the Vietnamese and Arabic community in Australia is questionable. Further studies designed specifically to investigate this are needed. PMID- 9226082 TI - The bereavement phenomenology questionnaire: a single factor only. AB - OBJECTIVE: To review the psychometric properties and factor structure of the Bereavement Phenomenology Questionnaire (BPQ). METHOD: Continuous data were compared using two tailed t-tests and analysis of variance. A confirmatory factor analysis was carried out. RESULTS: Good internal consistency (Cronbach's alpha = 0.93) with a single factor has been found. The BPQ discriminates the intensity of grief between widows and widowers, spouses and offspring, and temporally between acute and later phases of bereavement. CONCLUSION: Although on the BPQ items are highly correlated and only a single factor has emerged, it remains nonetheless a valid and reliable scale for the measurement of grief. PMID- 9226083 TI - Media depictions of mental illness: an analysis of the use of dangerousness. AB - OBJECTIVE: To explore how the commonsense understanding, that those with a mental illness are dangerous, is deployed in a small sample of print media. METHOD: The print media sample was subjected to a discourse analysis informed by knowledge of media practices. Materials were read closely and references to mental illness were identified, classified and analysed. RESULTS: This non-sensational material was shown to provide repeated confirmations of the commonsense understanding that mental illnesses make people unpredictable and dangerous. Close study of the lead article suggested that it was written so that readers had to draw on such understandings to make sense of the account it presented. CONCLUSION: The study challenges the notion that media present negative depictions of mental illnesses either because journalists are poorly informed or because 'sensation sells'. It is concluded that media practices directed at engaging readers require the use of cases and a style of writing that forces readers to draw upon commonsense knowledge of mental illness to understand the text. It is argued that this is a deliberate effort to enlist readers as co-creators of the text and thereby increase their interest. PMID- 9226084 TI - The stigma of mental illness in Asian cultures. AB - OBJECTIVE: This article reviews the attitudes towards mental illness and psychiatric stigma in Asian cultures. METHOD: Relevant literature published in English was reviewed. RESULTS: Psychiatric stigmas in Asian cultures share some common features. However, response to mental illness has many variations across cultures. Psychiatric stigma is prevalent and severe in some but not all Asian cultures. CONCLUSIONS: The stigma of mental illness needs to be studied within its sociocultural context in order to understand its origins, meanings and consequences. It may be relevant to examine the indigenous concepts, experience and implications of psychological problems to address problems in mental health care relating to stigma. PMID- 9226085 TI - Liaison psychiatry in an HIV/AIDS unit. AB - OBJECTIVE: To provide an overview of the work of a liaison psychiatry service to an HIV/AIDS inpatient unit, and particularly to examine the identification of mood and related disorders by referring doctors. METHOD: The MICRO-CARES prospective clinical database system was used to obtain data on all patients referred to the HIV/AIDS consultation-liaison psychiatry service in an infectious diseases hospital in Melbourne. RESULTS: Three hundred and ninety-two inpatient referrals were made in the 2 years from 1993-1995: a referral rate of 16.7%. The most frequent reasons for referral were evaluation of coping problems (42%), assessment of possible depression (31%), and assessment of psychotropic medication (24.5%). The most common psychiatric diagnoses were mood disorders (36.5%), psychoactive substance use disorders (22.7%) and organic mental disorders (18.1%). Overall concordance of recognition of depression by the referring doctor and diagnosis of depression by the consultant psychiatrist was 79%; 20% false positive rate, 23% false negative rate. CONCLUSIONS: Psychiatric comorbidity is common in patients with HIV/AIDS. Reasons for referral vary from those seen in other inpatient settings. Previously noted problems such as the misdiagnosis of psychiatric disorder and the mislabelling of the syndrome recognised by psychiatrists as depression were noted here. PMID- 9226086 TI - Experiences of physical and sexual abuse in Australian general practice attenders and an eating disordered population. AB - OBJECTIVE: To determine the reported rates of child physical and sexual abuse experienced by hospitalised eating disordered patients compared to a control group of women attending general practitioners. METHOD: A retrospective survey using the self-report Finkelhor Sexual Life Events Inventory and clinical reports. RESULTS: Nearly one-half of eating disordered patients reported a history of child sexual abuse and one-quarter reported child physical abuse. These rates were significantly higher than those reported by the control group. CONCLUSIONS: Direct questioning regarding trauma histories is warranted when assessing patients with eating disorders and attention to such issues should be incorporated into the total management plan. PMID- 9226087 TI - A survey of social outcome in schizophrenia in Tasmania. AB - OBJECTIVE: To survey the social outcome of patients with schizophrenia attending State mental health facilities in southern Tasmania. METHOD: Using the Statewide Mental Health Register, patients using inpatient and outpatient facilities who received a diagnosis of schizophrenia between 1981 and 1988 were identified (n = 771), and demographic and illness measures, and admissions and length of inpatient stay were compiled. The Life Skills Profile (LSP) was completed by mental health personnel for the 247 who were regular attenders or inpatients in 1991. RESULTS: Social morbidity as indexed by the LSP was highest in psychiatric hospital inpatients and patients in long-term rehabilitation programs, and lower in patients attending community centres. The majority of patients in suburban settings and attending community centres lived with their families, whereas patients in the inner city or in the rehabilitation service were mainly in hostel accommodation or living alone. Patients with schizophrenia attending State services were of a similar age range but had a longer duration of illness and more admissions, and had spent more days in hospital than patients who were not in regular contact with the service. CONCLUSIONS: The distribution of social morbidity in schizophrenia confirms that the public health system is supporting a group with high social morbidity. Patients with the highest morbidity are receiving the highest levels of care and intervention. PMID- 9226088 TI - Is clozapine safe in the elderly? AB - OBJECTIVE: The aim of this paper is to assess the risk/benefit ratio of clozapine in the elderly. METHOD: MedLine and Internet searches, followed by cross-checking for further articles, identified the references. Reports on efficacy as well as side effects were examined. Five psychogeriatric patients treated by the authors are presented. RESULTS: The review of the literature highlighted the lack of studies on the use of clozapine in the elderly. Retrospective studies and case reports dominate. There is only one double-blind placebo-controlled study of six patients extant. Although efficacy seemed to parallel that in the younger age groups, the incidence of leukopenia, agranulocytosis, postural hypotension and confusion was greater in the elderly. Two of the five patients reported by the authors developed agranulocytosis, bringing the Australian experience of agranulocytosis in patients over 65 years of age to 4/55. CONCLUSION: The risk/benefit ratio in the elderly is distinctly higher than found in the younger population. With specific reference to agranulocytosis alone, the frequently quoted risk of 1% for patients is likely to be 5-10 times higher in the over 65 population. Caution is advised when prescribing clozapine in the psychogeriatric population. PMID- 9226089 TI - Clozapine in the management of bipolar and schizoaffective manic episodes resistant to standard treatment. AB - OBJECTIVE: To test the efficacy of clozapine in treatment-resistant manic episodes. CLINICAL PICTURE: Three cases, two with bipolar disorder (manic) and one of schizoaffective disorder (manic), were treated with clozapine. TREATMENT: Clozapine was used after the failure of standard antipsychotics and mood stabilizers. OUTCOME: All three cases were successfully treated. CONCLUSION: A controlled trial of clozapine in treatment-resistant bipolar and schizoaffective manic episodes is indicated. PMID- 9226090 TI - Bilateral ulnar nerve paralysis: an unreported complication of drug-induced extrapyramidal rigidity. AB - OBJECTIVE: This case report describes a very unusual consequence of drug-induced extrapyramidal side effects. CLINICAL PICTURE: The patient developed bilateral ulnar nerve paralysis. TREATMENT: The treatment consisted of anticholinergic medication and physiotherapy. OUTCOME: The patient made a complete recovery over a period of 8 months. CONCLUSIONS: There is a need to ensure compliance with anticholinergic medication when using depot neuroleptic medication. PMID- 9226091 TI - Comment on Reconciling the patient's right to confidentiality and the family's need to know. PMID- 9226092 TI - Phenotyping for CYP 11D6 alleles. PMID- 9226093 TI - Comment on An epidemiological case for a separate adolescent psychiatry. PMID- 9226094 TI - WWW forensic psychiatry on-line. PMID- 9226095 TI - Comment on Neuroleptic malignant syndrome and clozapine monotherapy. PMID- 9226096 TI - Comment on Access to firearms and the risk of suicide: a case control study. PMID- 9226097 TI - Rate-dependent prolongation of action potential duration in isolated rat ventricular myocytes. AB - Rate-dependent alterations of action potential duration (APD) in rat ventricular myocytes were investigated. Action potentials of the isolated myocytes were recorded with patch electrodes containing EGTA (11 mM), and showed a marked rate dependent prolongation in the APD (0.2-5 Hz). This prolongation was significantly inhibited in the presence of 4-aminopyridine (4-AP), a blocker of the transient outward K+ current (Ito). Thus, the rate-dependent decrease in Ito may underlie the change in APD. In contrast, the action potentials recorded from rat ventricular papillary muscles with conventional microelectrodes did not show rate dependent alterations in the APD, i.e., the APD remained practically unaltered at the frequency range of 0.2-5 Hz. These results suggest that the rate-dependent prolongation of APD (due to rate-dependent blockade of Ito) becomes evident when the intracellular Ca2+ was chelated by the internal application of EGTA via patch pipette. We speculate that the rate-dependent prolongation of APD (via decreases in Ito) is masked in the ventricular papillary muscles, probably due to rate dependent decreases in the inward current (e.g., electrogenic Na(+)-Ca2+ exchange current) that is regulated by the intracellular calcium. PMID- 9226098 TI - Myocardial electrophysiological properties in the presence of an AT1 angiotensin II receptor antagonist. AB - INTRODUCTION: Blockade of the AT1 angiotensin II (Ang II) receptor has been shown to provide anti-hypertensive effects. However, whether AT1 Ang II receptor antagonists influence myocardial electrophysiological properties remains unclear. METHODS AND RESULTS: Accordingly, atrial and ventricular myocardial electrophysiological properties were examined in adult rat (n = 13) and guinea pig (n = 9) myocardial preparations in the presence of the specific AT1 Ang II receptor antagonist, valsartan (CGP 48933; 0.5, 5, or 500 mumol/L). These concentrations reflect up to 100 fold higher drug concentrations than those observed in clinical trials. Transmembrane potential data were recorded using standard microelectrode techniques at baseline and following superfusion with valsartan. The lower concentrations of valsartan (0.5 and 5 mumol/L) had minimal effects on myocardial electrophysiology. In the presence of 500 mumol/L of valsartan, resting membrane potential increased from baseline in both rat (-82.3 +/- 4.1 vs -76.8 +/- 5.8 mV, p < 0.05) and guinea pig (-81.6 +/- 2.9 vs -76.9 +/- 2.0 mV, p < 0.05) atrial myocardium. Action potential duration at 90% repolarization was increased in guinea pig atrial (91.7 +/- 1.4 vs 80.0 +/- 5.6 ms, p < 0.05) and ventricular (131.1 +/- 8.1 vs 118.7 +/- 8.3 ms, p < 0.05) myocardium following exposure to 500 mumol/L of valsartan. In a separate series of experiments, Ang II (1.0 mumol/L) had no effect on atrial or ventricular action potential characteristics in either species. CONCLUSION: Thus, the effects of valsartan, which were observed only at concentrations 100 fold higher than those reported in clinical trials, may be due to non-specific drug interactions with the myocyte sarcolemma. PMID- 9226099 TI - Myocardial angiotensin receptor type 1 gene expression in a rat model of cardiac volume overload. AB - To investigate the regulation of the angiotensin receptor type 1 (AT1) in different organs in cardiac volume overload, we measured AT1 mRNA content in the atria, left and right ventricle, kidney and liver of rats with an aortocaval shunt, produced by infrarenal aortocaval puncture 4 weeks earlier. For angiotensin receptor mRNA quantitation a novel quantitative PCR procedure based on liquid phase hybridization was used that allowed the determination of absolute AT1 mRNA copy numbers and its comparison in different organs. Glyceraldehydephosphate dehydrogenase (GAPDH) mRNA was measured by RT-PCR to control externally equal mRNA content and quality of RNA extraction in shunt animals and controls. Heart weight was increased in the shunt animals, with the greatest increase in the atria. Blood pressure, plasma renin activity, plasma angiotensin I and II and aldosterone concentrations were not significantly altered. The AT1 mRNA content was significantly increased in the atria (shunt: 1167 +/- 350 copies AT1 mRNA/ng RNA vs controls: 803 +/- 240; p < 0.05). No change was found in the right or left ventricle, in the kidney and liver. The findings document that atrial hypertrophy in cardiac volume overload parallel with a significant increase in atrial AT1 mRNA content. PMID- 9226100 TI - Functional consequences of acute collagen degradation studied in crystalloid perfused rat hearts. AB - OBJECTIVES: The impact of acute collagen disruption by the disulfide donor, 5,5' dithio-2-nitrobenzoic acid (DTNB) on ventricular properties was tested in rat hearts. METHODS: Collagen was degraded acutely in 13 isolated, isovolumically contracting rat hearts by perfusion with 1 mM DTNB added to Krebs-Henseleit solution for 1 hour followed by 2-hour perfusion with normal solution. Another 13 hearts were perfused with normal solution for 3 hours (Control). RESULTS: Collagen content was 3.5 +/- 0.5% of ventricular dry weight in control group compared with 2.1 +/- 0.4% in DTNB group (decrease by 40%, p < 0.01). Scanning electron micrographs revealed loss of the delicate collagen network surrounding muscle fibers in DTNB treated hearts. Developed pressure at a fixed volume decreased to 86 +/- 17% of the baseline value after 3-hour perfusion in the control group, whereas in DTNB treated hearts developed pressure fell to 68 +/- 13% (p < 0.01). End-diastolic pressure was set at 5 mmHg at the beginning of the experiment and rose to 15 +/- 8 mmHg in control and 30 +/- 13 mmHg (p < 0.01) in the treated hearts. Concomitantly, wet-to-dry weight ratio increased from 5.63 +/ 0.26 in control to 6.07 +/- 0.11 (p < 0.05) in the DTNB treated hearts. A separate set of experiments on isolated myocytes excluded the possibility of a direct effect of DTNB on myocyte contractile function. CONCLUSIONS: These data suggested that with 40% collagen disruption by DTNB there is a significant increase in tissue edema that results in a decrease in chamber capacitance; in addition, there is a significant decrease in systolic performance which reflects the combined effect of edema and loss of collagen. PMID- 9226101 TI - Characterisation of the infarct-limiting effect of delayed preconditioning: timecourse and dose-dependency studies in rabbit myocardium. AB - The delayed phase ('second window') of protection induced by ischemic preconditioning in rabbit heart is observed as enhanced resilience to infarction 24 hours after repetitive brief cycles of ischemia. Here we provide a fuller physiological characterisation of this phenomenon in the open-chest rabbit model, examining temporal characteristics and dose-dependency of this adaptation. For examination of the timecourse of delayed protection, rabbits were pretreated with four 5 minute coronary artery occlusions (PC) or sham operation (SHAM). Twenty four, 48, 72 or 96 hours later, infarct size after 30 min coronary occlusion and 120 minutes reperfusion was assessed with TTC staining and expressed as a percentage of myocardial risk volume (I/R). I/R was reduced at 24 hours (SHAM 48.1 +/- 3.9% v PC 31.4 +/- 3.0%, P < 0.01), 48 hours (SHAM 41.9 +/- 3.0% v PC 19.6 +/- 6.3%, P < 0.01), and 72 hours (SHAM 39.8 +/- 3.4% v PC 17.2 +/- 2.5%, P < 0.01). No protection was observed 96 hours after preconditioning (SHAM 35.0 +/- 4.8% v PC 36.9 +/- 3.8%). In a further study, animals were pretreated with one, two or four 5 minute coronary occlusions (1 x 5 PC, 2 x 5 PC, 4 x 5 PC) and subjected to the infarction protocol 48 hours later. I/R was 44.5 +/- 4.3% in SHAM, 24.8 +/- 4.4% in 1 x 5 PC (P < 0.01), 27.4 +/- 2.9% in 2 x 5 PC (P < 0.05) and 24.4 +/- 4.8 in 4 x 5 PC (P < 0.01). Delayed protection in this rabbit model is prolonged, extending between 24 and 72 hours after the preconditioning stimulus. The threshold for eliciting the second window of protection in this model is as low as one 5 minute coronary occlusion. PMID- 9226102 TI - Mechanical properties and effects of sympathetic co-transmitters on human coronary arteries and veins. AB - Active isometric wall tension was studied at different levels of passive wall tension in isolated circular 2 mm long segments of human epicardial coronary arteries and veins, and maximum active wall tension was calculated to 6.60 mN/ mm for arteries and 0.86 mN/mm for veins. Vasomotor responses to sympathetic co transmitters were studied at resting tension and after precontraction with U46619. Noradrenaline (NA) and adenosine 5'-triphosphate (ATP) induced strong contractions of veins, whereas relaxant responses dominated in arteries. Isoprenaline potently relaxed all arteries and veins. Prazosin and rauwolscine in a concentration of 10(-7) M both competitively antagonized NA-induced contraction of arteries and veins. For uridine 5'-triphosphate (UTP), relaxant responses were demonstrated in most arteries but only some veins. Neuropeptide Y (NPY) elicited no observable vasomotor responses in either arteries or veins. Mechanical removal of the arterial endothelium did not significantly alter relaxant responses to NA, ATP, UTP or isoprenaline. In conclusion, alpha 1- and alpha 2-adrenoceptors mediating contraction and beta-adrenoceptors mediating relaxation seem to be present in both human epicardial coronary arteries and veins. When applied to isolated epicardial coronary vessels, NA and ATP had a stronger influence on vasomotor tone than NPY and UTP, mediating strong contraction of veins but mainly relaxation of coronary arteries, that was independent of an intact endothelium. PMID- 9226103 TI - DTTX30, a combined thromboxane receptor antagonist and thromboxane synthetase inhibitor, prevents coronary thrombosis in anesthetized dogs. AB - BACKGROUND: Combined thromboxane A2 receptor blockade and thromboxane synthetase inhibition facilitates local prostacyclin production at the site of platelet activation thereby providing a potent antithrombotic effect. The efficacy of DTTX30, a combined thromboxane A2 receptor blocker-thromboxane synthetase inhibitor, in inhibiting recurrent coronary thrombosis was evaluated in vivo using a canine model of unstable angina pectoris. METHODS AND RESULTS: Pentobarbital-anesthetized dogs (total of 25) were used in which acute damage of the proximal left circumflex coronary artery, together with mechanical stenosis, produced an occlusive thrombosis. When the platelet plug was removed by rubbing the vessel, the occlusion returned reproducibly for at least 2 hours in control studies. To evaluate the antithrombotic efficacy of DTTX30, a reproducible pattern of occlusive coronary thrombi was first established over a period of one hour. Thereafter, DTTX30 (0.12 mg/kg i.v. bolus plus 0.29 mg/kg/hr), acetylsalicylic acid (ASA, 5 mg/kg, i.v. bolus), vapiprost (1.0 mg/kg i.v. bolus plus 1.0 mg/kg/ hr), or vehicle was administered and observations continued for one additional hour. At the end of the one hour observation period, epinephrine (0.3 microgram/kg/min i.v.) was infused (with continued DTTX30, vapiprost or vehicle infusion) and observations were continued for an additional 30 min. DTTX30 eliminated the recurrent arterial thrombus formation in all dogs without significant systemic or left ventricular hemodynamic effects. Infusion of epinephrine to further provoke thrombus formation produced a significant enhancement of left ventricular pressure development (LV dP/dt) and arterial diastolic and systolic pressures but failed to initiate thrombus formation. DTTX30 inhibited fully collagen-induced platelet aggregation ex vivo and produced an approximate 3-fold prolongation of the sublingual bleeding time. ASA eliminated thrombus formation in 4 of 5 dogs, but the additional prothrombotic stimulus of epinephrine infusion produced recurrent thrombus formation in all dogs. Collagen-induced platelet aggregation ex vivo was inhibited by acetylsalicylic acid, but sublingual bleeding time was unaffected. Vapiprost inhibited arterial thrombus formation in all dogs, but thrombus formation returned with the addition of epinephrine. There was a tendency for prolonged bleeding times with vapiprost administration and collagen-induced platelet aggregation ex vivo was effectively inhibited. CONCLUSION: These studied indicate that the combined inhibition of the thromboxane A2 receptor together with inhibition of thromboxane synthetase provides superior antithrombotic activity in vivo than does thromboxane A2 receptor blockade alone (vapiprost) or inhibition of cyclooxygenase (ASA). Thrombus formation even with the additional stimulation of epinephrine was inhibited by DTTX30 but neither vapiprost nor ASA was effective in this setting. PMID- 9226104 TI - Validation of the multiple colored microsphere technique for regional blood flow measurements in newborn piglets. AB - The use of multiple colored microspheres (CMS) for the measurement of regional blood flow (RBF) in almost all organs and tissues of newborn piglets was validated. For this purpose mixtures of different CMS and/or radio-labeled microspheres (RMS) were injected into the left ventricle of eight newborn piglets. Regional blood flows (RBF) were quantified using the reference sample method. Flow rates estimated by RMS and CMS were compared for each tissue sample. An excellent correlation (r = 0.995-0.999) between CMS and RMS flow rates was found even for organs with low perfusion and tissue samples containing 400-750 CMS. We conclude that the CMS technique is a valid alternative for RBF measurement in newborn piglets, and that all disadvantages arising from radioactive labeling are thereby avoided. PMID- 9226105 TI - Digoxin and its related endogenous factors. AB - The digitalis drugs are plant-derived cardenolide compounds used medicinally for several hundred years. These drugs elicit inotropic and chronotropic effects on the heart, but they also affect many other tissues. The mechanism of action involves inhibition of the ion-transport activity of a membrane-associated protein called Na, K-ATPase (sodium pump). Present theory holds that the sodium pump is the principal molecular receptor for the digitalis drugs. Recent evidence indicates the presence of naturally occurring digitalis-like compounds in mammals. It is believed these compounds, collectively known as either digitalis like (DLF) or ouabain-like (OLF) factors, may be endogenous hormones regulating the biological activity of the sodium pump and its isoforms. The presence of deglycosylated and other congeners of one specific DLF, the digoxin-like immunoreactive factor (DLIF), has very recently been described in humans. Digoxin as a drug is the most widely prescribed digitalis in the U.S., and its measurement in serum has established a model for present-day therapeutic drug monitoring (TDM). Historically, the accurate measurement of digoxin in blood has been difficult. This article focuses on the present understanding of the clinical use of digoxin, factors that affect the accuracy of measuring digoxin, the principle of measuring metabolically active species of digoxin, and the effects of DLIF and other interfering substances in digoxin immunoassay. PMID- 9226106 TI - Current concepts and advances in clinical laboratory testing for autoimmune diseases. AB - This review discusses the current concepts of immunological tolerance, physiological vs. pathological autoimmunity, autoimmune diseases, and laboratory tests helpful in diagnosis. The autoantibodies in organ-specific autoimmune diseases are directed against antigens of the injured organs, whereas the antinuclear antibodies (ANA) detected in systemic autoimmune diseases are detected against a vast array of nuclear and intracellular antigens and peptides necessary for DNA/RNA synthesis, repair, splicing, and transcription. Knowledge of the mean titer and presence or absence of specific ANA types will help predict the nature of the disease and the response to therapy. Noteworthy features of these "ANA profiles" are (1) patients with systemic lupus erythematosus frequently have multiple types of ANA but anti-dsDNA and anti-SM are diagnostic, (2) patients with drug-induced lupus have ANA restricted to antihistone, (3) patients with mixed connective tissue disease have ANA restricted to anti-RNP, (4) patients with CREST (calcinosis, Raynaud's, esophageal dysmotility, sclerodactyly, and telangiectasia) syndrome have ANA restricted to anticentromere, (5) ANA with anti-SS-A/Ro specificity is associated with vasculitis and nephritis, (6) ANA with anti-SS-B/La and anti-nRNP specificities is associated with milder clinical disease, (7) ANAs with anti-Jo-1 and PM-Scl specificities are associated with pulmonary fibrosis and poor prognosis. Technological advances in the fields of molecular immunogenetics are guiding the studies of autoimmune diseases from serological and histopathological evaluations toward search for subcellular risk factors such as chemical and biological agents and susceptibility genes. Knowledge of these factors will help (1) to identify disease susceptibility genes prior to clinical onset and irreversible tissue damage, (2) to avoid environmental risk factors, and (3) to devise specific immunosuppressive strategies. PMID- 9226108 TI - Strategies for imaging biliary neoplasms. AB - Biliary tract neoplasms are rare, develop in either intra- or extrahepatic locations, and are represented by a spectrum of imaging characteristics. These tumors may be imaged using a variety of traditional methods, such as ERCP, PTC, ultrasound, or CT, and some newer innovative cross-sectional techniques, such as MR- or CT-cholangiography. The choice of imaging technique depends on whether the study is for diagnostic, staging, or therapeutic purposes. Because these tumors are frequently small, secondary features such as intrahepatic ductal dilatation or even hepatic metastases, may be the initial finding on screening studies. Benign tumors, postsurgical strictures, or even biliary sludge may mimic features of malignant tumors, and the radiologist must be aware of these potential sources of interpretative error. PMID- 9226107 TI - Liver MRI with contrast enhancement. AB - Liver MR imaging has improved substantially in the last decade due to advances in equipment design and pharmaceutical development. Contrast agents for the liver have been examined within three major classes. Extracellular agents (Gd HP-DO3A, Gd DTPA, and Gd DTPA-BMA) are currently in widespread clinical use. Gadolinium chelates with hepatobiliary excretion are in clinical trials (Gd BOPTA and Gd EOB DTPA). With respect to particulate agents, one (AMI-25) has received Food and Drug Administration (FDA) approval recently. Imaging technique is critical to proper use, with different strategies employed depending on the class of agent. With gadolinium chelates, T1-weighted images are used for visualization, with dynamic scans being important for extracellular agents, and both dynamic and delayed scans being important for agents with hepatobiliary excretion. With the particulate iron compounds, T2-weighted images are used for visualization, with emphasis on delayed imaging. PMID- 9226109 TI - Antibodies to Listeria monocytogenes. AB - Listeria monocytogenes is one of the leading foodborne pathogens and has been implicated in numerous outbreaks in the last 2 decades. Immunocompromised populations are usually the most susceptible to Listeria infections. Although the pathogenic mechanism is a complex process, significant progress has been made in unravelling the mechanism in recent years. It is now clear that numerous extracellular and cell-associated proteins, such as internalin, listeriolysin, actin polymerization protein, phospholipase, metalloprotease, and possibly p60 proteins, are essential for L. monocytogenes entry into mammalian cells, survival inside the phagosome, escape into the cytoplasm, and cell-to-cell spread. Other proteins may be responsible for growth and physiology or to maintain the structural integrity of the bacteria. Monoclonal and polyclonal antibodies have been developed against many of those antigens or their synthetic derivatives that have helped greatly to determine the structure and function of these antigens. The antibodies were also used for the diagnosis and detection, immunocytochemical staining, and serotyping of Listeria. Humoral immune response to live L. monocytogenes cells was examined in naturally or experimentally infected hosts. Studies revealed that only extracellular antigens induced the humoral response, whereas cell-associated antigens had apparently no response. It is speculated that during the occasional bacteremic phase, L. monocytogenes releases extracellular antigens that are then processed by the immune system for antibody production. As L. monocytogenes is an intracellular pathogen, the cell-associated antigens are not persistent in the blood circulation and thus fail to stimulate the humoral immune response. PMID- 9226110 TI - Public health and nonpasteurized fruit juices. AB - Well publicized outbreaks of foodborne illness have occurred in recent years due to consumption of commercial, nonpasteurized ("fresh" or "unpasteurized") fruit juices. Nonpasteurized and heat treated juices have been associated with at least 15 foodborne illness outbreaks since the early 1900s. Disease syndromes have included salmonellosis, typhoid fever, cyrptosporidiosis, Escherichia coli related diarrhea, and hemolytic uremia. Mortality has occasionally occurred during these outbreaks. An increase in the number of reported outbreaks in recent years possibly reflects greater consumption of fresh juices and closer scrutiny of these products by medical and public health authorities. This article reviews the fruit juice borne outbreaks in the 1900s, methods to control pathogens, and regulatory issues related to production of nonpasteurized fruit juices in the U.S. PMID- 9226111 TI - Immunologic epitope, gene, and immunity involved in pneumococcal glycoconjugate. AB - Pneumococcal infection persists as a major cause of pneumonia, otitis media, and meningitis in infants. Children less than 2 years of age show the highest incidence of pneumococcal diseases. Production of monoclonal antibody (MAb) to polysaccharide (PS) and binding characteristics to PS epitopes were studied. Removal of the O-acetyl group from 9V PS by alkali hydrolysis resulted in a decreased binding with rabbit 9V antiserum (AS). However, the binding reaction with 9V MAb was less affected by the loss of O-acetyl content. Type 9V IgG MAb provided passive protection and enhanced the opsonophagocytic activity of polymorphonuclear (PMN) leukocytes to kill type 9V pneumococci. The pathogenecity of pneumococci is attributed to various virulence factors distributed on the cell surface, including capsular polysaccharide and protein antigens, for example, pneumolysin, autolysin, pneumococcal surface protein A (PspA), pneumococcal surface adhesion (PsaA), and hemin binding protein. Some of these protein antigens may be used as a component to combine with pneumococcal PS vaccine or as a carrier of conjugate vaccine. Clinical trials of pneumococcal conjugate vaccines showed that covalent linkage of capsular PS to protein carriers improved the immunogenicity of the PS. Development of glycoconjugate vaccine for selected pneumococcal types will help solve the problem of poor immunogenecity of PS vaccine in young children used for prevention of pneumococcal infection. PMID- 9226112 TI - Natural protection of spring and well drinking water against surface microbial contamination. I. Hydrogeological parameters. AB - The fate and transport of microbes in groundwater are controlled by physicochemical characteristics of the microbe and of the groundwater/aquifer media. Key characteristics of the microbe include size, inactivation (die-off) rate, and surface electrostatic properties. Key properties of the groundwater/aquifer system include flow velocity, aquifer grain (or pore) size, porosity, solid organic carbon content, temperature, pH, and other chemical characteristics of water and mineral composition. Because of size and surface electrical properties, viruses are much more mobile in groundwater than Cryptosporidium and Giardia (which are about 100 times or more larger than viruses). The inactivation or die-off rate is usually the most important factor governing how far microbes can migrate in significant numbers in groundwater. Typical half-lives of microbes in groundwater range from a few hours to a few weeks. Examples of maximum reported migration distances of microbes in groundwater include: bacteria, 600 m in a sandy aquifer: viruses, 1000 to 1600 m in channeled limestones and 250 to 408 m in glacial silt-sand aquifers; Cryptosporidium and Giardia, no confirmed reports found of significant migration distances. Investigations by the EPA have indicated that distances of 210 to 325 m away from septic tanks are necessary to achieve with high confidence an 11 order of magnitude reduction in virus concentrations. PMID- 9226114 TI - Does change in skin perfusion provide a good index to monitor the sympathetic response to a noxious stimulus in preterm newborns? AB - Skin perfusion was measured using laser Doppler fluximetry (LDF) in 16 preterm babies undergoing a standardised heel prick procedure. Although there was a significant reduction in skin blood flow following the heel prick, this was variable and dependent on basal skin blood flow. This, together with loss of data due to movement artefact, makes this technique unreliable in quantifying the sympathetic response to a noxious stimulus in preterm infants. PMID- 9226113 TI - Natural protection of spring and well drinking water against surface microbial contamination. II. Indicators and monitoring parameters for parasites. AB - Recent outbreaks of cryptosporidiosis and reports of other newly described para sitic diseases associated with drinking water transmission prompted a reevaluation of source water monitoring criteria for public health protection. The field of microbial indicators was reviewed and each candidate sentinel evaluated in terms of its sensitivity, specificity, and technical feasibility. In addition, a clear distinction was made between source water monitoring and monitoring in the distribution system. Of all potential candidate microbial sentinels, Escherichia coli is deemed the most efficacious for public health protection. Based on a conservative estimate of its half-life in groundwater for 8 d, it is recommended that at least two samples be obtained during this half life. In addition to E. coli, two water quality indicator sentinels, which are not necessarily direct public health threats, should also be monitored at the same frequency. These are the total coliform group and the enterococci. If E. coli is present in any source water sample, the borehole and any directly connected borehole should be embargoed. If either total coliforms or enterococci are detected, only that individual borehole should be taken off line and not used until the situation is remediated and the cause of the fecal contamination eliminated. Clostridium perfringens spores serve as a useful long-lived indicator. However, their perseverance in a sample should not be considered a direct public health threat because spores may far outlive pathogens. As a parasite indicator, C. perfringens should have the same importance as a positive coliform or enterococcus analysis. Coliphages do not yet fulfill enough of the criteria to be routinely employed. Biological monitoring should be coupled with physicochemical monitoring to establish a long-term history of the source. Because all natural waters vary in the amounts of heterotrophic plate count bacteria, test methods should be employed that are refractory to them. A combination of rigorous source protection plus extraordinary source monitoring serve as sufficient multiple barriers for parasite protection. PMID- 9226115 TI - Transvaginal fetal biometry in early pregnancy. AB - OBJECTIVE: To produce reference charts for fetal size with transvaginal sonography that are potentially helpful in evaluating normal and abnormal early pregnancies. DESIGN: A prospective cross-sectional study. SUBJECTS: 1081 normal singleton pregnancies with a normal fetal karyotype or normal healthy baby at delivery, at 9-16 weeks' gestation. Measurements included crown rump length, biparietal diameter, transverse cerebellar diameter, head and abdominal circumference, mean abdominal diameter, thoracic circumference, femur length, humerus length and foot length. RESULTS: The best description of the relationship between single ultrasonographic parameters and gestational age was achieved by polynomial regression analysis. All fetal biometric parameters correlated closely with gestational age. Biparietal diameter maintained the closest correlation with gestational age (r2 = 97.15, p < 0.001; y = -0.545 - 0.06x + 0.15x2); transverse cerebellar diameter showed the poorest correlation with gestational age (r2 = 88.17, p < 0.001). Reference ranges (5 degrees and 95 degrees percentile intervals) were constructed for all biometric parameters in relation to gestational age. Mean residuals are similar for all parameters with a very low range. CONCLUSIONS: These data provide normograms for first and early second trimester fetal measurements which may be of aid in the dating of pregnancies and can be useful in the early detection of genetic disorders affecting the growth of fetal structures. PMID- 9226116 TI - Levels of amniotic fluid insulin and profiles of maternal blood glucose in pregnant women with diabetes type-I. AB - The aim of this study was to investigate the relationship between amniotic fluid insulin (AF-insulin) measurements and maternal blood glucose levels in pregnancies complicated by insulin-dependent maternal diabetes mellitus (IDDM). Twenty-five patients with IDDM underwent amniocentesis (AC) in the third trimester. Twelve patients had a second amniocentesis after 2-3 weeks. The maternal blood glucose values (MBG) 2 weeks before amniocentesis were correlated with AF-insulin. Mean (+/-S.D.) MBG in the group with AF-insulin > 97th centile (n = 7) was 6.1 +/- 1 mmol/l. MBG in the group with AF-insulin < 97th centile (n = 18) was 5.3 +/- 1.2 mmol/l (r = 0.2948; P-value 0.162). In the group with repeated AC and AF-insulin > 97th centile (n = 6) the correlation coefficient was 0.722 (P = 0.043), whereas in the group with normal AF-insulin (n = 6) no correlation was found (r = -0.213; P = 0.686). These results indicate that no significant correlation exists between MBG values and concentration of AF insulin. MBG is not appropriate for the diagnosis of fetal hyperinsulinism in well-controlled women with IDDM. In individual cases with AF-insulin > 97th centile a decrease of MBG causes lower AF-insulin levels. These results indicate that there seems to be an individual threshold for maternal MBG which causes hyperinsulinism. Fetal hyperinsulinism not only depends on blood glucose levels. Different fetal sensitivity to maternal glucose stimuli or a different glucose transport across the placenta in the individual fetus could be responsible for these results. PMID- 9226117 TI - Non-invasive blood pressure measurements and aortic blood flow velocity in neonates. AB - We studied the systolic blood pressure difference between the upper and the lower extremities in healthy newborn infants and the effect of the isthmic narrowing of the aorta on the possible difference. The blood pressure was measured with an oscillometric blood pressure device from every extremity of 36 healthy infants aged 2-5 days. A Doppler echocardiography was performed for each infant to measure the aortic blood flow velocity in the ascending aorta and in the aortic arch above and below the isthmic narrowing. The mean blood pressure readings (S.D.) were the following: the right arm 76.8 (7.3)/48.1 (6.9), the left arm 77.5 (7.4)/51.6 (7.0), the right thigh 77.7 (7.1)/40.7 (5.8), the left thigh 76.8 (6.4)/39.6 (5.8), the right calf 75.5 (7.1)/46.6 (5.7) and the left calf 77.1 (8.6)/48.7 (6.7). The aortic blood flow was faster below the isthmic narrowing of the aorta (1.15 +/- 0.19 m/s) than in the ascending aorta (0.93 +/- 0.12 m/s) or in the aortic arch above the isthmus (0.99 +/- 0.15 m/s). The calculated pressure gradient between the ascending aorta and aorta below the isthmus was 2.0 +/- 1.8 mmHg and between opposite sides of the isthmus 1.5 +/- 1.2 mmHg. Unlike in childhood and adolescence, the systolic blood pressure in the lower extremities of healthy newborn infants is not higher than in the upper extremities. The physiological narrowing of the aortic arch does not explain this phenomenon. If blood pressure measurements are performed on a neonate to rule out aortic coarctation, the readings obtained must be interpreted in respect to normal values in newborns. PMID- 9226118 TI - Ultrasound growth parameters in relation to levels of amniotic fluid insulin in women with diabetes type-I. AB - The aim of the study was to investigate the correlation between ultrasound parameters and levels of amniotic fluid insulin (AF-insulin) in pregnancies complicated by insulin-dependent diabetes mellitus (IDDM). In 129 women with IDDM amniocentesis was performed between 28 and 35 weeks of gestation. The levels of AF-insulin were measured by radioimmunoassay (Pharmacia RIA 100) and were correlated with biparietal diameter (BPD), abdominal diameter (AD), abdominal circumference (AC), and femur length (FL). The women were maintained at good glycemic control (fructosamine level: mean +/- S.D.: 236.3 +/- 40 micromol/l) and delivered infants with a mean (+/- S.D.) birth weight of 3477 +/- 640 g. The sensitivity of BPD, AD, AC and FL to detect fetuses with pathological levels of AF-insulin was 50%, 62%, 67% and 49%, respectively. The sensitivities of AD and AC in a selected group (n = 14) with highly pathological levels of AF-insulin (> 20 microU/ml) were both 80%, whereas the specificity was 56% and 46%, respectively. In women with IDDM, fetal biparietal diameter, abdominal diameter, abdominal circumference, and femur length are not reliable markers for the identification of fetal hyperinsulinism. Only cases with highly pathological levels of AF-insulin can be detected by abdominal measurements. PMID- 9226119 TI - Temporal disparity between reduction of cot death and reduction of prone sleeping prevalence. AB - According to several reports sudden infant death rates have decreased significantly after public campaigns aimed at reducing the incidence of sleeping in a prone position. The Styrian population (1.2 million inhabitants), who have been studied from 1984, also showed a significant drop in the incidence of cot death during 1989 (from 2/1000 to 1/1000%). The year before, a campaign for the prevention of cot death had been launched. This included the recommendation to prevent infants from lying in a prone position during sleep. Part of the prevention programme consisted of a detailed questionnaire filled in and returned by the parents. These data, on 29970 infants from 1989 to 1994, provided information on the frequency of prone sleeping in 37% of our total population and as a consequence on parental response to the campaign. Calculating the data per year led to the surprising result that the reduction by half (from 50% to 25%) in the prevalence of sleeping in a prone position did not occur in 1989, when the drop in the incidence of cot death occurred, but 3 years later, in 1992. The following years saw a further decrease of prone position to 7% but no appreciable change in the incidence of cot death. However, during those 11 years of study about 80% of the victims were consistently found dead lying in a prone position. Our results show a temporal disparity between the reduction of sudden infant death and the decrease of prone sleeping in a population. Although we do not deny sleeping in a prone position as a risk factor for cot death, there cannot be a simple relationship between sleeping habits in the population and incidence of cot death. PMID- 9226120 TI - Lyso-phosphatidylcholine and outcome of preterm babies with respiratory distress syndrome treated with surfactant. AB - Phosphatidylcholine (PC) is the predominant phospholipid in natural surfactant preparations. A metabolic intermediate, lyso-PC, is potentially injurious to the lungs. In the present study, tracheal aspirates from preterm babies with respiratory distress syndrome treated with surfactant were examined for the presence of lyso-PC to determine if there was any correlation with outcome. Eighteen babies were assigned to receive initially either 100 or 200 mg/kg Curosurf followed by up to three further 100-mg/kg doses if required. Lyso-PC was present in aspirates taken 12-24 h after the last treatment from nine of 11 infants who initially received 200 mg/kg but in only one from seven receiving 100 mg/kg initially, and was dependent on the total dose of phospholipid administered. Three babies in the low-dose group developed bronchopulmonary dysplasia, whereas two in the high-dose group were non-survivors, however we could not correlate the presence of lyso-PC with adverse long-term outcome in this group of preterm infants. PMID- 9226121 TI - Is the age of menopause determined in-utero? AB - OBJECTIVE: To determine whether age at menopause is related to size at birth. DESIGN: A follow-up study of two groups of women whose size at birth was recorded. SETTING: Hertfordshire and Sheffield, England. POPULATION: 755 women aged 60-71 years born in Hertfordshire; 235 women aged 40-42 years born in the Jessop Hospital, Sheffield. MAIN OUTCOME MEASURES: Age at natural menopause or serum follicle stimulating hormone concentration greater than 25 IU/ml. RESULTS: Age at menopause was unrelated to birth weight. However, it occurred at a younger age in women who had low weight at 1 year. This was independent of their body weight and smoking habits. In the population of younger women those who had had an early menopause tended to have been short at birth, with a high ponderal index (birth weight/length3). CONCLUSION: Growth retardation in late gestation, leading to shortness at birth and low weight gain in infancy, may be associated with a reduced number of primordial follicles in the ovary leading in turn to an earlier menopause. PMID- 9226122 TI - Stability of urinary HVA and VMA on filter paper. AB - The study examined the stability of HVA and VMA in 1-ml aliquots of a single urine sample stored on filter paper at different temperatures for 2 years. The results showed that HVA and VMA were stable in dried filter paper when stored at 4 degrees C or lower temperature. Storage at room temperature resulted in degradation of the sample. PMID- 9226123 TI - Possible sex-correlated transmission of maternal class I HLA haplotypes. AB - Forty-seven alleles of class I HLA-AB loci (14 for locus A and 33 for locus B) were identified in 787 participants in two groups of unrelated families. Group I included parents and children typed for bone marrow transplantation. Group II included families typed for renal transplantation. Before statistical evaluation, the A locus alleles were grouped into eight classes according to broad specificity, and the B locus alleles were grouped according to HLA epitopes into two classes. Significant differences in HLA-AB haplotype frequencies were found between male and female offspring. When families with children of both sexes were analysed, the frequencies of maternally inherited HLA-AB haplotypes were found to be significantly different in brothers and sisters. The results suggest the possibility that the transmission of specific AB haplotypes from mother to offspring may be correlated to children's sex. The major histocompatibility complex has been shown to be involved in the expression of H-Y male-specific minor histocompatibility antigens. The possible selection in the transmission of specific maternal HLA-AB haplotypes to male offspring may contribute to the avoidance of maternal cytotoxic reactions toward the male foetus. PMID- 9226124 TI - HLA-DQ associations and T-cell receptor V-gene usage in peripheral blood of Swedish myasthenia gravis patients. AB - To characterize better the functional aspects of the HLA class II associations with myasthenia gravis (MG), T-cell receptor (TCR) V alpha/beta elements were studied in peripheral blood in 29 Swedish MG patients. HLA typing had previously been done using polymerase chain reaction with sequence-specific primers (PCR SSP) or combined with sequence-specific oligonucleotide probes (PCR-SSO). The TCR V gene expression was determined by fluorescence-activated cell sorter (FACS) analysis using 12 monoclonal antibodies (mAb) that detected 30-40% of CD4+ and CD8+ T cells. No correlation between HLA-DQ genotype and TCR V elements could be found, nor was any restricted V gene usage seen. Fourteen (48%) of the patients had T cells showing signs of abnormal expansion in peripheral blood. There was an increased expression of TCR V gene elements in CD8+ T cells in patients (13/29) compared with CD4+ T cells in patients (5/29) (P < 0.05) and in unthymectomized patients compared with controls (14/56) (P < 0.005). TCR V gene expression was also increased in the CD8+ population in unthymectomized (7/8) compared with thymectomized patients (6/21) (P < 0.01). There was an increased expression in both CD4+ and CD8+ populations in unthymectomized patients (7/8, 88%), compared with thymectomized patients (7/21) (P < 0.05). We conclude that the abnormal T cell expansion in peripheral blood could be a reflection of non-specific pathogenic processes in the muscle and thymus. PMID- 9226126 TI - Allelic polymorphism of two multifunctional regions in the central human MHC: tenascin X, XB-S and YB, and their duplicated fragments XA and YA. AB - Two highly polymorphic sequences have been discovered in the complement C4 region of the human major histocompatibility complex (MHC). They are part of a duplicated unit of overlapping genes, transcribed in opposite directions and containing the sequences of tenascin X (XB), XB-S, XA, YB and YA. Fragments of 1014 bp and 894 bp were co-amplified by polymerase chain reaction (PCR) and digested with two different restriction enzymes. This preliminary study provides evidence for more than five different alleles each of XA (YA) and XB (XB-S, YB), and at least 11 XA-XB haplotypes. Their association with extended HLA-B-DR haplotypes, C4 complotypes, and C4 region restriction fragment length polymorphisms (RFLP) is discussed. X gene polymorphisms are a complement region marker system that should be uniquely suited for PCR-based typing methods. They could become a useful addition to HLA class I and class II markers in the mapping of candidate genes for MHC-associated diseases, including the X and Y genes themselves. PMID- 9226125 TI - Report from the HLA class II typing by PCR-SSP Multicentre Study. AB - Results from 360 HLA-DR and -DQ 'low-resolution' typings with polymerase chain reaction sequence-specific primers (PCR-SSP), performed by nine laboratories, were analysed for their overall utility in routinely defining the HLA-DR1-DR18, DR51-DR53 and DQ1-DQ9 specificities in less than 2.5 h. Thirty EDTA blood samples and 10 DNA samples were distributed and analysed by each laboratory. DNA was extracted using a rapid bromide salt extraction protocol. Complete HLA-DR and -DQ typings were performed, three by three, on pre-aliquoted 96-tube PCR trays. When compared with reference typing, 351/360 (98%) correct DR typings were obtained, whereas 320/360 (89%) of the DQ phenotypes were correctly assigned. The time for three complete HLA-DR and -DQ 'low-resolution' typings, including DNA extraction, ranged from 2.0 h to 2.3 h. Unfortunately, an unusually high level of PCR amplification failures was observed (3%), probably due to diffusion and a significant volume loss from some of the pre-aliquoted primer mixes. Consequently, only 52% of the typings were without any amplification failure, and 0-2 amplification failures where found in 88% of the PCR-SSP typings performed. The number of HLA-DR-DQ retypings needed was 7 and 8%, respectively, reflecting the low number of typings where allelic identification was directly affected by the relatively high level of amplification failures in this study. Thus, a 91-98% success rate of correctly identified HLA-DR and -DQ alleles could be maintained, even under suboptimal typing conditions. PMID- 9226127 TI - The specificity of anti-HLA class II monoclonal antibodies in cattle. AB - At the Eleventh International HLA Histocompatibility Workshop, numerous anti-HLA class II monoclonal antibodies (mAb) were tested. For several of the polymorphic mAb, one epitope for binding has been mapped within the antigen-binding site of the class II molecules. Screening of the available bovine DRB3 and DQB exon 2 sequences revealed that some of the key amino acid (AA) motifs of these epitopes were present in cattle as well, and the question was raised whether this sharing of key AA motifs might cause interspecies cross-reactivity. Eight polymorphic anti-HLA class II mAb (seven anti-HLA DRB1 and one anti-HLA DQB) were selected for analysis of their reactivity towards bovine lymphocytes. In addition, the monomorphic anti-HLA class II mAb, 7.5.10.1, was selected for analysis, as this mAb was described to detect class II polymorphism in cattle. Flow cytometry and lymphocyte microcytotoxicity testing revealed that five of the polymorphic anti HLA mAb were reactive with bovine lymphocytes. Furthermore, the anti-bovine reactivity of 7.5.10.1 was confirmed. These findings were supported by biochemical analysis. The anti-bovine reaction of the anti-HLA mAb did not correspond with the expected reaction, which was based on the presence of the AA, postulated to be responsible for recognition. Therefore, we suggest that the patterns of reactivity of the anti-HLA mAb are not always determined by one epitope. PMID- 9226128 TI - Sequence of a new HLA-DR allele, DRB5*0106. AB - We report here the exon 2 sequence of a new HLA-DRB5 allele, DRB5*0106, that was identified in two volunteer bone marrow donors from the Swiss national registry. This new allele differs from DRB5*0101 by five amino acids at positions 67, 70, 71, 85 and 86. It is associated with DRB1*1501 and DQB1*0602. This unusual DRB1*1501-DRB5*0106 association increases the complexity of the DR2 group, although it appears to be very rare, at least in our population. HLA-DRB5*0106 can be readily detected upon DR generic oligotyping, provided the two probes that mark the major DRB5 subtypes, DRB5*0101 and DRB5*02, respectively, are included in the assay. PMID- 9226129 TI - Polymorphisms of TAP1 and TAP2 genes in German patients with type 1 diabetes mellitus. AB - Type 1 diabetes mellitus (IDDM) is an autoimmune disorder in which the alleles HLA DQA1*0501-DQB1*0201 and DQA1*0301-DQB1*0302 confer strong susceptibility. The genes for transporters associated with antigen processing (TAP1 and TAP2) are located near HLA DQ and display only a limited degree of polymorphism. Since polymorphisms of TAP might influence susceptibility to IDDM possibly by selection of different antigen peptides, we investigated sequence variants of TAP1 and TAP2 genes in 120 German patients with IDDM and 218 random healthy German controls by polymerase chain reaction (PCR) followed by sequence-specific oligonucleotide analysis (SSO), single-strand conformation polymorphism (SSCP) analysis and amplification refractory mutation system (ARMS). TAP1*02011 (16% vs. 4% in controls, P = 0.001, RR = 5.0) and TAP2*0101 (96% vs. 69% in controls, P < 0.0001, RR = 10.6) showed a positive association with IDDM. However, these associations disappeared when patients and controls were matched for predisposing HLA DQA1 or DQB1 alleles as well as for DRB1*0401. In conclusion, our findings indicate that the observed association of TAP variants with IDDM in German patients is due to linkage disequilibrium with HLA DQ alleles/DRB1*04 subtypes. PMID- 9226130 TI - Nomenclature for factors of the HLA system, update January/February 1997. PMID- 9226131 TI - Nomenclature for factors of the HLA system, update March 1997. PMID- 9226132 TI - Propiomelanocortin (POMC) gene expression by normal skin and keloid fibroblasts in culture: modulation by cytokines. AB - Originally described as a product of the pituitary gland, propiomelanocortin (POMC) has recently been identified in other tissues, such as in human skin, where it may accumulate in response to various stimuli. Thus far, epidermal keratinocytes, as well as melanocytes and macrophages, have been shown to express POMC. This study investigated the expression of POMC mRNA in cultured dermal fibroblasts derived from either normal skin or keloids. Using Northern blot hybridization with a POMC cDNA generated by RT-PCR of mRNA isolated from cardiac muscle, we demonstrated that dermal fibroblasts express POMC, as significant levels of mRNA were detected in unstimulated cells in culture. POMC transcript steady-state levels were strongly reduced by transforming growth factor-beta (TGF beta), whereas tumor necrosis factor-alpha (TNF-alpha) counteracted the effect of TGF-beta and exerted a stimulatory activity on POMC mRNA levels. Reduction of POMC transcript levels by TGF-beta was also observed in cultured keratinocytes. Clearly detectable levels of POMC mRNA were detected in cultured keloid-derived fibroblasts; however, little, if any, regulation by TGF-beta was observed. These data represent the first demonstration of POMC expression by fibroblasts and down regulation by TGF-beta. Furthermore, our results indicate altered TGF-beta regulation of POMC gene expression in keloid-derived fibroblasts, suggesting that POMC may play a role in the pathogenesis of keloid formation. PMID- 9226133 TI - Epidermal localization and protective effects of topically applied superoxide dismutase. AB - Data from the literature, as well as our previous work, indicate a protective effect of superoxide dismutase (SOD) in topical application against UV-induced cutaneous damage. In the present article we show that pre-treatment of the skin with SOD protects against PUVA-induced inflammatory reactions not only in murine, but also in human skin. Using fluorescently labelled Cu,Zn SOD, epifluorescence microscopy and digital image processing, we demonstrate that the FITC fluorescence localizes in the stratum corneum and upper granulosa, as well as in the epidermal cell layer surrounding the lumina of the hair follicles. These findings were similar for murine and human skin. Since autofluorescence was eliminated by a special filter, it can be ascertained that the fluorescence observed in the tissues was due to FITC-labelled SOD. PMID- 9226134 TI - A realistic approach to the sensitivity of PCR-DGGE and its application as a sensitive tool for the detection of clonality in cutaneous T-cell proliferations. AB - The practical value of the detection of clonality within the T-cell receptor gamma locus by polymerase chain reaction for the diagnosis of cutaneous T-cell lymphomas is well known. However, studies dealing with this subject so far, with special emphasis on the sensitivity of the technique in comparison to, for example, Southern blotting have used mixtures of DNA in various concentrations instead of using mixtures of the cells involved, which would reflect the in vivo situation in a more realistic scope. The purpose of this study was therefore to determine the sensitivity and the limitations of the PCR assay by dilution experiments, using mixtures of cells. Furthermore we studied its applicability to cutaneous T-cell proliferative disorders. Two clonal T-cell lines (MyLa and Jurkat) served as positive control. Dilutions of MyLa cells, cultured normal human keratinocytes and peripheral blood mononuclear cells from lymphoma negative volunteers were used to assess the sensitivity of the PCR-DGGE assay. Skin samples from 4 patients with cutaneous T-cell lymphoma, 1 lesional lymph node, 2 blood samples from a patient with Sezary syndrome and 4 lymphoma-negative tissue samples were analysed. Two samples were uncertain for diagnosis of lymphoma. The PCR-DGGE assay consisted of a 2-round nested PCR with consensus primers within the TCR-gamma locus followed by electrophoretic separation of the product along a denaturing urea/formamide gradient gel. PCR-DGGE sensitivity was, to our knowledge, for the first time investigated for mixtures of lymphocytes (clonal and polyclonal) and keratinocytes. Clonal T-cells were detected in a concentration between 1-0.1% in keratinocytes, whereas the sensitivity was generally lower upon dilution in peripheral blood mononuclear cells or in a mixture of keratinocytes and peripheral blood mononuclear cells. Nevertheless, T cell clonality was detected in 2 blood samples of a patient with Sezary syndrome, which were negative by Southern blot analysis. The crucial point of this work was the new approach to establish the sensitivity of the PCR-DGGE, in a way which more closely mimics the condition of clinical specimens. Instead of mixing and amplifying DNA extracted from clonal T-cell lines and polyclonal bone marrow cells, we amplified DNA from clonal and polyclonal cells which had been mixed in various ratios before DNA extraction. Polymerase chain reaction in conjunction with denaturing gradient gel electrophoresis is a sensitive and versatile molecular tool for the assessment of clonality of suspect cutaneous lesions. The determination of sensitivity using DNA extracted from premixed cells more closely corresponds to the actual test situation when testing skin samples. PMID- 9226135 TI - beta-Adrenergic stimulation induces activation of protein kinase C and inositol 1,4,5-trisphosphate increase in epidermis. AB - Epidermal keratinocytes express beta 2-adrenergic receptors on the cell membrane. The binding of the agonists to the beta 2-adrenergic receptors regulates activation of adenylate cyclase. This transmembrane signaling system has been regarded to be one of the important pathways for the functions of keratinocytes. We previously reported that beta-adrenergic stimulation induced a transient increase of intracellular Ca2+ in normal human epidermal keratinocytes. Thus we investigated the effects of epinephrine on another transmembrane signaling system, the phosphatidyl-inositol signal transduction pathway in pig epidermis. Treatment of pig pure epidermis with epinephrine resulted in a transient increase in inositol 1,4,5-trisphosphate with a peak at 30 s. Epinephrine induced translocation of protein kinase C from cytosol to the membrane fraction. The activation of protein kinase C, translocation of protein kinase C from cytosol to the membrane fraction, was confirmed using the beta-adrenergic agonist isoproterenol. Moreover, the effect of epinephrine on the activation of protein kinase C was inhibited by preincubation with propranolol, a beta-adrenergic antagonist. The increase in inositol 1,4,5-trisphosphate and translocation of protein kinase C by epinephrine are consistent with the view that beta-adrenergic stimulation induces turnover of inositol phospholipid in pig epidermis. PMID- 9226136 TI - Molecular analysis of different phases in human wound healing. AB - Cultured granulation fibroblasts grown from punch biopsies of the same lower arm area, obtained 3, 6, 9 and 14 days after wounding, were used as a human wound healing model in comparison to quiescent fibroblasts. We investigated the expression of key extracellular matrix components at the protein level by flow cytometry and mRNA steady state levels by Northern blotting of the different fibroblasts and compared these data to the ability to migrate towards a chemotactic signal. Procollagen alpha 1 (I), fibronectin and matrix metalloprotease-1 synthesis was strongly up-regulated at the mRNA steady state level on days 3 and 14. Tissue inhibitor of metalloprotease-1 mRNA is only 20% down-regulated between day 3 and 14. Chemotaxis towards conditioned medium reflects a net effect of several factors and is distinctly different from chemotaxis towards platelet-derived growth factor, which peaks at day 3. Compared to the protein level, the enhanced expression of the corresponding PDGF receptor beta chain mRNA is delayed by 3 to 6 days. PDGF receptor alpha shows no regulatory changes during the observation period. This data further supports the idea that functionally divergent subpopulations of fibroblasts exist during wound healing. PMID- 9226137 TI - Isolation of a cDNA encoding a tyrosine kinase expressed in murine skin. AB - Tyrosine phosphorylation is widely recognized as playing an important role in cell differentiation, proliferation and carcinogenesis. We used the polymerase chain reaction (PCR) method to identify protein tyrosine kinases that were expressed in the skin. Mixed oligonucleotide probes were used to amplify and screen neonatal murine skin mRNA for clones encoding amino acid contiguities, the conservation of which is characteristic of the protein tyrosine kinase family. When the PCR products were sequenced, a novel clone encoding protein tyrosine kinase, PTK70, was identified. A full-length cDNA was isolated from a mouse thymus cDNA library. The nucleotide and deduced amino acid sequence showed that it featured src-homology (SH) 2 domain, SH3 domain and kinase domain like other src family protein tyrosine kinases, but lacked the N-terminal myristylation site and C-terminal tyrosine residue. Although the mRNA of PTK70 was detected in various tissues ubiquitously, the degree of its expression differed among tissues. Murine skin is one in which PTK70 was expressed strongly, with its expression being much stronger in the epidermis and in the cell line derived from murine keratinocytes than in those from melanoma or fibroblast cell lines. These evidences suggest that PTK70 may be involved in proliferation or differentiation of keratinocytes in the skin. PMID- 9226138 TI - Serum-free conditions for the long term growth and differentiation of neoplastic canine keratinocytes. AB - Long term cultures of canine keratinocytes have been established but culture conditions currently used require supplementation with fetal bovine serum (FBS). Unfortunately, FBS contains many non-defined components which may interfere with in vitro studies. This study describes the development of defined serum-free culture conditions for neoplastic canine keratinocytes grown submerged and at the air-liquid interface. Two commercially available serum-free media established for human epidermal cells failed to support canine keratinocyte growth. In contrast, a defined serum-free medium developed in our laboratory successfully supported proliferation of neoplastic canine keratinocytes for at least 40 passages. Cells showed a slower growth rate, but reached similar final densities and were morphologically identical to those cultured in FBS. Grown at the air-liquid interface, the cells reached the same degree of differentiation as in vivo stratified squamous epithelium and cultures grown in FBS. These results demonstrate that canine keratinocytes require different serum-free growth conditions than human cells. Neoplastic canine keratinocyte cultures, grown under serum-free culture conditions, provide an ideal in vitro system for comparative studies of keratinocyte biology and pathogenesis of various dermatoses. PMID- 9226139 TI - Distal posterior interosseous nerve syndrome. AB - Five patients presenting with chronic dorsal wrist pain and diagnosed as distal posterior interosseous nerve syndrome are reported. Clinical examination revealed point tenderness of the fourth extensor compartment. The symptoms were reproduced by extreme wrist extension in all patients and by extreme flexion in three of the patients. The pain was relieved in all patients by a selective lidocaine block of the terminal branch of the posterior interosseous nerve at the wrist joint. All the patients failed to respond to nonoperative treatment and underwent surgical exploration of the nerve. The operative findings were an enlarged nerve in three patients and adhesion of the nerve to the joint capsule in five patients. A 2 cm section of the nerve proximal to the extensor retinaculum was resected. Four of the five patients had excellent pain relief and returned to full asymptomatic activity. One patient had improvement in pain and was satisfied with the outcome. A diagnosis of distal posterior interosseous nerve syndrome should be considered if the usual sources of dorsal wrist pain are eliminated. PMID- 9226140 TI - Modelling in economic evaluation: an unavoidable fact of life. AB - The role of modelling in economic evaluation is explored by discussing, with examples, the uses of models. The expanded use of pragmatic clinical trials as an alternative to models is discussed. Some suggestions for good modelling practice are made. PMID- 9226141 TI - Statistical inference for cost-effectiveness ratios. AB - Methods for statistical inference for cost-effectiveness (C/E) ratios for individual treatment and for incremental cost-effectiveness (delta C/ delta E) ratios when two treatments are compared are presented. In a lemma, we relate the relative magnitude of two C/E ratios to the delta C/ delta E ratio. We describe a statistical procedure to test for dominance, or admissibility, that can be used to eliminate an inferior treatment. The one-sided Bonferroni's confidence interval procedure is generalized to the two-sided case. The method requires only that two confidence intervals be available, one for cost and one for effectiveness. We describe Fieller-based confidence intervals and show them to be shorter than Bonferroni intervals. When distribution assumptions hold and variance and covariance estimates are available, Fieller intervals are preferable. However, Bonferroni intervals can be applied in more diverse situations and are easier to calculate. A simple Bonferroni based technique, and a likelihood ratio statistic given by Siegel, Laska and Meisner, for testing the null hypothesis that the C/E ratios of two treatments are equal is presented. The approaches are applied to the data from a phase II clinical trial of a new treatment for sepsis considered previously by others. PMID- 9226142 TI - Confidence intervals for cost-effectiveness ratios: a comparison of four methods. AB - We evaluated four methods for computing confidence intervals for cost effectiveness ratios developed from randomized controlled trials: the box method, the Taylor series method, the nonparametric bootstrap method and the Fieller theorem method. We performed a Monte Carlo experiment to compare these methods. We investigated the relative performance of each method and assessed whether or not it was affected by differing distributions of costs (normal and log normal) and effects (10% absolute difference in mortality resulting from mortality rates of 25% versus 15% in the two groups as well as from mortality rates of 55% versus 45%) or by differing levels of correlation between the costs and effects (correlations of -0.50, -0.25, 0.0, 0.25 and 0.50). The principal criterion used to evaluate the performance of the methods was the probability of miscoverage. Symmetrical miscoverage of the intervals was used as a secondary criterion for evaluating the four methods. Overall probabilities of miscoverage for the nonparametric bootstrap method and the Fieller theorem method were more accurate than those for the other the methods. The Taylor series method had confidence intervals that asymmetrically underestimated the upper limit of the interval. Confidence intervals for cost-effectiveness ratios resulting from the nonparametric bootstrap method and the Fieller theorem method were more dependably accurate than those estimated using the Taylor series or box methods. Routine reporting of these intervals will allow individuals using cost effectiveness ratios to make clinical and policy judgments to better identify when an intervention is a good value for its cost. PMID- 9226143 TI - Productivity costs measurement through quality of life? A response to the recommendation of the Washington Panel. AB - This paper comments on the recently published guidelines of the Washington Panel on incorporation of indirect non-medical costs, or productivity costs, in economic evaluations of health care. Traditionally the human capital or more recently the friction cost method is used to measure these costs. The Panel, however, recommends incorporating productivity costs as health effects in the denominator of the C/E ratio. This paper argues that incorporation of productivity costs in cost-effectiveness analysis expressed as health effects is not correct. Only direct health related effects on quality of life that cannot be meaningfully monetarized should be considered as health effects. Furthermore, measuring productivity costs in terms of quality of life may lead to misrepresentation of these costs from a societal viewpoint. This misrepresentation occurs because of the existence of social security systems and private insurance compensating for income reductions from disease. Furthermore, the patient's viewpoint is useful for quality of life measurement, but not for measuring productivity costs from a societal perspective. Finally, alternative recommendations are formulated for incorporating societal productivity costs in economic evaluations of health care. PMID- 9226144 TI - Using conjoint analysis to assess women's preferences for miscarriage management. AB - To date, standard gamble, time trade-off, visual analogue and, more recently, willingness to pay, have been most commonly employed in health economics to assess utilities from various health care interventions. This article considers the use of conjoint analysis as an alternative technique to assess utilities. The technique is applied to assess women's preferences for the management of miscarriage. The paper addresses methodological issues in the application of the technique to health care and demonstrates its use in estimating willingness to pay and utilities. It is concluded that conjoint analysis is potentially a very useful tool and that future research should investigate more thoroughly the potential application of the technique in health economics. PMID- 9226145 TI - Economic evaluation and the shifting balance towards primary care: definitions, evidence and methodological issues. AB - Current UK government policy places considerable emphasis on shifting the balance of health care provision from secondary towards primary care. Despite this emphasis, however, there has been very little economic evaluation of initiatives designed to achieve this shift. In view of this deficiency, our article has three main aims. First, it offers a working definition of shifts in the balance of care at the primary-secondary care interface. Second, it systematically reviews the existing literature on the cost-effectiveness of initiatives designed to shift the balance of care. Third, it identifies a range of methodological issues that need to be addressed if future economic evaluations in this area are to be carried out satisfactorily. PMID- 9226146 TI - Output efficiency of health maintenance organizations in Florida. AB - We use data envelopment analysis (DEA) to measure the relative technical efficiencies of 28 HMOs licensed to practice in the State of Florida in the autumn of 1994. Health care output measures used in the analysis are number of commercial, Medicare and Medicaid members enrolled in each plan. Inputs to the model are capital assets, total expenditures on the provision of medical services and administrative expenses. We find differences in HMO efficiency scores and loss ratios (defined as the ratio of expenses on the provision of medical services to the total expenses incurred by the organization) across individual plans. Differences in efficiency measures across model type (staff, IPA, combination) and ownership types (for-profit, not-for-profit) are small but significant: staff models and for-profits are more efficient. In a multivariate model, we also find that large HMOs are more efficient and HMOs with Medicaid patients are significantly less efficient than other HMOs. PMID- 9226147 TI - Equity in health care utilization: further tests based on hurdle models and Swedish micro data. AB - This paper tests the null hypothesis of no horizontal inequity in delivery of health care by use of count data hurdle models and Swedish micro data. It differs from most earlier work in three principal ways: First, the tests are carried out separately for physician and hospital care; second, the tests are carried out separately for the probability of seeking care and the amount of care received (given any use); and third, the tests are based on a model that includes several socioeconomic variables, e.g. income, education and size of community of residence. The paper rejects the hypothesis of no inequity because socioeconomic factors also have significant effects on utilization, e.g. income and size of community of residence. Size of community of residence has a positive significant effect on the frequency of physician visits but not on the probability of visiting a physician. PMID- 9226148 TI - Juvenile myelomonocytic leukemia. PMID- 9226149 TI - Transcription factors, normal myeloid development, and leukemia. PMID- 9226150 TI - Abnormal expression of the B-cell homing chemokine receptor BLR1 during the progression of acquired immunodeficiency syndrome. AB - The putative chemokine receptor BLR1 has been identified as the first G-protein coupled receptor involved in B-cell migration and in microenvironmental homing to B-cell follicles and to germinal centers. In healthy individuals, expression of BLR1 is restricted to all mature recirculating B cells and to a subpopulation of T-helper memory cells. In the present study, we analyzed the distribution of BLR1 on defined lymphocyte subsets during the progression of acquired immunodeficiency syndrome. It is shown that the proportion of T-helper memory cells coexpressing BLR1 continuously decreases during the infection, whereas a high proportion of gamma/delta T cells expressing BLR1 can be found in peripheral blood. The latter subpopulation is restricted to lymphoid tissues in healthy individuals. Most interestingly, in 75% of all human immunodeficiency virus (HIV)+ individuals, peripheral blood B cells were identified as not expressing BLR1 and phenotypically resembling germinal center cells of lymphoid tissue. Using BLR1 as a marker molecule, this study identifies peripheral blood lymphocytes in HIV+ individuals that are usually restricted to lymphoid tissue in healthy individuals. Because HIV infection is active in lymphoid tissue even at the clinically latent stage, aberrant expression of the B-cell homing chemokine receptor BLR1 might be an early indicator for the onset of destruction of lymphoid tissue. PMID- 9226151 TI - The Ig heavy chain gene is frequently involved in chromosomal translocations in multiple myeloma and plasma cell leukemia as detected by in situ hybridization. AB - Chromosome rearrangement of 14q32.33 has recurrently occurred with variable partner sites, including 11q13.3, 8q24.1, 18q21.3, and 6p21.1 in multiple myeloma (MM). To assess the actual incidence of 14q32.33 translocation and to elucidate its implication in the pathogenesis of MM, we studied 42 patients with MM, plasma cell leukemia, or plasmacytoma and 5 with monoclonal gammopathy with undetermined significance (MGUS) by G-banding and molecular cytogenetic methods. Using double color fluorescence in situ hybridization (DCFISH) with 2 Ig heavy chain (IgH) gene probes, a yeast artificial chromosome (YAC) clone containing variable region, and a phage clone containing gamma constant region, 14q32.33 translocation was detected as split signals of the IgH gene in 31 patients with plasma cell malignancies and 3 with MGUS. In contrast, of 40 patients who were assessed by G-banding, 3 (7.5%) showed the 14q+ chromosome. DCFISH detected a split of the IgH gene on interphase nuclei in 34 (73.9%) of 46 patients analyzed, whereas on metaphase spreads, it was in 22 (51.2%) of 43 patients analyzed. Interphase DCFISH was particularly useful to detect 14q32.33 translocation in 17 (65.4%) of 26 patients with normal karyotypes. Donor sites were identified in 11 of 22 patients demonstrated as carrying 14q32.33 translocation by metaphase FISH. Chromosome t(11;14)(q13.3; q32.33) was detected in 5 patients, t(8;14)(q24.1;q32.33) in 2, t(14;18)(q32.33;q21.3) in 2, and t(7;14)(q32.1;q32.33) in 1. A complex 14q32.33 translocation involving 3q and 16q24 was detected in 1 patient. Myeloma cells with t(7;14) showed myelomonocytoid surface antigen. Because rearrangements of 14q32.33 were closely associated with translocation of proto-oncogenes into the IgH gene, our findings indicate that 14q32.33 translocation with various partner chromosomes is a critical event in the pathogenesis of MM and MGUS. PMID- 9226152 TI - All patients with the T(11;16)(q23;p13.3) that involves MLL and CBP have treatment-related hematologic disorders. AB - The involvement of 11q23-balanced translocations in acute leukemia after treatment with drugs that inhibit the function of DNA topoisomerase II (topo II) is being recognized with increasing frequency. We and others have shown that the gene at 11q23 that is involved in all of these treatment-related leukemias is MLL (also called ALL1, Htrx, and HRX). In general, the translocations in these leukemias are the same as those occurring in de novo leukemia [eg, t(9;11), t(11;19), and t(4;11)], with the treatment-related leukemias accounting for no more than 5% to 10% of any particular translocation type. We have cloned the t(11;16)(q23;p13.3) and have shown that it involves MLL and CBP (CREB binding protein). The CBP gene was recently identified as a partner gene in the t(8;16) that occurs in acute myelomonocytic leukemia (AML-M4) de novo and rarely in treatment-related acute myeloid leukemia. We have studied eight t(11;16) patients, all of whom had prior therapy with drugs targetting topo II with fluorescence in situ hybridization (FISH) using a probe for MLL and a cosmid contig covering the CBP gene. Both probes were split in all eight patients and the two derivative (der) chromosomes were each labeled with both probes. Use of an approximately 100-kb PAC located at the breakpoint of chromosome 16 from one patient revealed some variability in the breakpoint because it was on the der(16) in three patients, on the der(11) in another, and split in four others. We assume that the critical fusion gene is 5'MLL/3'CBP. Our series of patients is unusual because three of them presented with a myelodysplastic syndrome (MDS) most similar to chronic myelomonocytic leukemia (CMMoL) and one other had dyserythropoiesis; MDS is rarely seen in 11q23 translocations either de novo or with t-AML. Using FISH and these same probes to analyze the lineage of bone marrow cells from one patient with CMMoL, we showed that all the mature monocytes contained the fusion genes as did some of the granulocytes and erythroblasts; none of the lymphocytes contained the fusion gene. The function of MLL is not well understood, but many domains could target the MLL protein to particular chromatin complexes. CBP is an adapter protein that is involved in regulating transcription. It is also involved in histone acetylation, which is thought to contribute to an increased level of gene expression. The fusion gene could alter the CBP protein such that it is constitutively active; alternatively, it could modify the chromatin-association functions of MLL. PMID- 9226153 TI - A metalloproteinase inhibitor prevents lethal acute graft-versus-host disease in mice. AB - Tumor necrosis factor (TNF) and Fas ligand (FasL) have been implicated in the pathogenesis of graft-versus-host disease (GVHD), which is a major complication after allogeneic bone marrow transplantation. We examined here the ameliorating effect of a metalloproteinase inhibitor (KB-R7785) that inhibits TNF-alpha and FasL release in a lethal acute GVHD model in mice. Administration of KB-R7785 into (BALB/c x C57BL/6) F1 that received C57BL/6 spleen cells markedly reduced the mortality and weight loss in association with minimal signs of GVHD pathology in the liver, intestine, and hematopoietic tissues. The ameliorating effect of KB R7785 was superior to that of anti-TNF-alpha antibody. Our results suggest that KB-R7785 could be a potent therapeutic agent for GVHD. PMID- 9226154 TI - Ex vivo generation of human anti-pre-B leukemia-specific autologous cytolytic T cells. AB - In contrast to other neoplasms, antigen-specific autologous cytolytic T cells have not been detected in patients with human pre-B-cell leukemias. The absence of efficient B7 family (B7-1/CD80; B7-2/CD86) -mediated costimulation has been shown to be a major defect in tumor cells' capacity to function as antigen presenting cells. We show here the generation of autologous anti-pre-B-cell leukemia-specific cytolytic T-cell lines from the marrows of 10 of 15 patients with pre-B-cell malignancies. T-cell costimulation via CD28 is an absolute requirement for the generation of these autologous cytolytic T cells (CTL). Although costimulation could be delivered by either bystander B7 transfectants or professional antigen-presenting cells (indirect costimulation), optimal priming and CTL expansion required that the costimulatory signal was expressed by the tumor cell (direct costimulation). These anti-pre-B-cell leukemia-specific CTL lysed both unstimulated and CD40-stimulated tumor cells from each patient studied but did not lyse either K562 or CD40-stimulated allogeneic B cells. Cytolysis was mediated by the induction of tumor cell apoptosis by CD8+ T cells via the perforin-granzyme pathway. Although we were able to generate anti-leukemia specific CTL from the bone marrow, we were unable to generate such CTL from the peripheral blood of these patients. These studies show that antigen-specific CTL can be generated from the bone marrow of patients with pre-B-cell leukemias and these findings should facilitate the design of adoptive T-cell-mediated immunotherapy trials for the treatment of patients with B-cell precursor malignancies. PMID- 9226155 TI - Comparative activity of melarsoprol and arsenic trioxide in chronic B-cell leukemia lines. AB - Inorganic arsenic trioxide (As2O3) was recently shown to induce apoptosis in NB4 promyelocytic leukemic cells. We have investigated the effects of the organic arsenical, melarsoprol (a drug used for treatment of trypanosomiasis), upon induction of apoptosis in cell lines representative of chronic B-cell lymphoproliferative disorders. An Epstein-Barr virus (EBV)-transformed B prolymphocytic cell line (JVM-2), an EBV-transformed B-cell chronic lymphocytic leukemia (B-CLL) cell line (I83CLL), and one non-EBV-transformed B-CLL cell line (WSU-CLL) were used as targets. Dose-response experiments with melarsoprol (10( 7) to 10(-9) mol/L) were performed over 96 hours. Unexpectedly, we found that melarsoprol caused a dose- and time-dependent inhibition of survival and growth in all three cell lines. In contrast, As2O3 at similar concentrations had no effect on either viability or growth. After 24 hours, all three cell lines treated with melarsoprol (10(-7) mol/L) exhibited morphologic characteristics of apoptosis. We also observed prominent concentration-dependent downregulation of bcl-2 mRNA after 24 hours of exposure to melarsoprol in WSU-CLL, I83CLL, and JVM 2 cells. Decrease of bcl-2 protein expression was also observed in all three cell lines, whereas As2O3 had no effect on this parameter. We conclude that melarsoprol may inhibit the growth of lymphoid leukemic cell by promoting programmed cell death. Results of these studies suggest that melarsoprol shows promising therapeutic activity in these diseases, and a study to evaluate clinical effects of this drug has been initiated. PMID- 9226156 TI - Incidence and clinical relevance of TEL/AML1 fusion genes in children with acute lymphoblastic leukemia enrolled in the German and Italian multicenter therapy trials. Associazione Italiana Ematologia Oncologia Pediatrica and the Berlin Frankfurt-Munster Study Group. AB - The molecular approach for the analysis of leukemia associated chromosomal translocations has led to the identification of prognostic relevant subgroups. In pediatric acute lymphoblastic leukemia (ALL), the most common translocations, t(9;22) and t(4;11), have been associated with a poorer clinical outcome. Recently the TEL gene at chromosome 12p13 and the AML1 gene at chromosome 21q22 were found to be involved in the translocation t(12;21)(p13;q22). By conventional cytogenetics, however, this chromosomal abnormality is barely detectable and occurs in less than 0.05% of childhood ALL. To investigate the frequency of the molecular equivalent of the t(12;21), the TEL/AML1 gene fusion, we have undertaken a prospective screening in the running German Berlin-Frankfurt-Munster (BFM) and Italian Associazione Italiana Ematologia Oncologia Pediatrica (AIEOP) multicenter ALL therapy trials. We have analyzed 334 unselected cases of pediatric ALL patients consecutively referred over a period of 5 and 9 months, respectively. The overall incidence of the t(12;21) in pediatric ALL is 18.9%. The 63 cases positive for the TEL/AML1 chimeric products ranged in age between 1 and 12 years, and all but one showed CD10 and pre-B immunophenotype. Interestingly, one case displayed a pre-pre-B immunophenotype. Among the B lineage subgroup, the t(12;21) occurs in 22.0% of the cases. Fifteen of 61 (24.6%) cases coexpressed at least two myeloid antigens (CD13, CD33, or CDw65) in more than 20% of the gated blast cells. DNA index was available for 59 of the 63 TEL/AML1 positive cases; a hyperdiploid DNA content (> or = 1.16) was detected in only four patients, being nonhyperdiploid in the remaining 55. Based on this prospective analysis, we retrospectively evaluated the impact of TEL/AML1 in prognosis by identifying the subset of B-lineage ALL children enrolled in the closed German ALL-BFM-90 and Italian ALL-AIEOP-91 protocols who had sufficient material for analysis. A total of 342 children were investigated for the presence of TEL/AML1 fusion gene and 99 cases (28.9%) were positive. The patients expressing the TEL/AML1 fusion mRNA appeared to have a better event-free survival (EFS) than the patients who lacked this chimeric product. Whereas three of the TEL/AML1 positive cases (3.0%) have relapsed to date, 27 patients without TEL/AML1 rearrangement (11.1%) suffered from relapse. To date, the only subset of B-lineage ALL with a favorable prognosis has been the hyperdiploid group (DNA index > or = 1.16 < 1.6). Our findings reinforce the need to include the molecular screening of the t(12;21) translocation within ongoing prospective ALL trials to prove definitively its prognostic impact. PMID- 9226157 TI - Increased frequency of expression of elevated dihydrofolate reductase in T-cell versus B-precursor acute lymphoblastic leukemia in children. AB - The relationships between dihydrofolate reductase (DHFR) levels or methotrexate membrane transport and acute lymphoblastic leukemia (ALL) immunophenotype were evaluated on 51 T-cell and 44 B-precursor ALL specimens from 90 pediatric ALL patients at diagnosis and relapse, using a fluorescent methotrexate analog (PT430) and flow cytometry assay (Matherly et al, Blood 85:500, 1995). For T-cell ALL, 35 of 45 (78%) of newly diagnosed patients' specimens exhibited elevated DHFR relative to DHFR levels in ALL blasts from methotrexate-responsive patients. For 30 of 45 diagnostic T-ALL specimens, DHFR expression was heterogeneous, with up to 3 separate subpopulations covering a 44-fold range; the DHFR-overproducing fractions comprised 10% to 88% of the total blasts. Elevated DHFR was less common in B-precursor ALL at diagnosis, being detected in only 17 of 36 specimens (47%); 11 of these samples exhibited DHFR heterogeneity. Median maximal DHFR levels were fourfold higher in T-cell than B-precursor ALL at diagnosis. Within a particular phenotypic group, there were no correlations between DHFR levels and patient prognostic features, including age, sex, chromosomal abnormalities, white blood cell counts (WBCs), and percentage of S-phase. However, for B-precursor ALL, there was a notably higher fraction of African-American than white patients with elevated DHFR. For patients (both phenotypes) with low WBCs (<50,000/ microL), event-free survival times were significantly shorter for those expressing DHFR above a threshold level than for patients expressing DHFR below this level (P < .016); this relationship was not seen for patients with high WBCs (>50,000/microL). Elevated DHFR was detected in 11 of 14 relapse specimens (5 of 6 T-cell and 6 of 8 B-precursor). Two of five paired relapse specimens (both T cell) from patients treated with methotrexate exhibited increased DHFR levels over those at diagnosis (2.5- to 5-fold); the fraction of DHFR-overproducing blasts was also increased in 4 of 5 paired relapse specimens (2 B-precursor and 2 T-cell). In contrast to the variations in DHFR, highly impaired methotrexate transport was detected in only 6 of 95 ALL specimens, including both diagnosis and relapse. There was no correlation between phenotype and impaired transport. These data provide further rationale for the use of mechanistically based prognostic factors to complement known biologic or disease-based prognostic indicators in the design of ALL therapy including methotrexate, particularly with patients presenting with low WBCs. The finding of a markedly increased frequency of DHFR overexpression in T-cell over B-precursor ALL suggests that this difference is associated with the poorer prognosis of patients with T-cell ALL treated with standard-dose antimetabolite therapy and implies that higher-dose methotrexate (> or = 1 g/m2) during consolidation therapy may be useful in the treatment of this disease. PMID- 9226158 TI - Granulocyte colony-stimulating factor as an adjunct to induction chemotherapy for adult acute lymphoblastic leukemia--a randomized phase-III study. AB - Because of the recommendation to avoid the concomitant administration of growth factors and chemotherapy, there is only limited information on colony-stimulating factor (CSF) therapy in acute lymphoblastic leukemia (ALL) induction protocols, in which cytotoxic drugs are administered in divided doses over a prolonged period of time, thus requiring a simultaneous administration of growth factors and chemotherapy. We conducted a prospective, randomized, controlled study to determine the safety and efficacy of granulocyte colony-stimulating factor (G CSF; filgrastim) as an adjunct to phase I of induction chemotherapy for adult ALL. Patients (n = 53) were randomized to receive no growth factor or G-CSF (5 microg/kg/d subcutaneously) starting on day 2 of chemotherapy consisting of daunorubicin (45 mg/m2) and vincristine (1.5 mg/m2) on days 1, 8, 15, and 22; L asparaginase (2500 U/m2) on days 1 through 14; and prednisone (60 mg/m2) on days 1 through 28. A total of 25 patients in the G-CSF group and 26 patients in the control arm fulfilled the inclusion criteria of the study. G-CSF markedly ameliorated neutropenia because the median proportion of days with neutropenia less than 1,000/microL was 29% in the G-CSF group as compared with 84% in the control arm (P < .00005). The median time to reach absolute neutrophil counts (ANC) > or = 1,000/microL was 16 days in G-CSF patients and 26 days in controls (P < .001). More importantly, G-CSF significantly reduced the incidence of febrile neutropenia (12% v 42% in controls, P < .05) and documented infections (40% v 77%, P < .05). No significant differences were found with regard to requirements for red blood cell transfusions and platelet concentrates. A total of 24 of 25 (96%) patients in the G-CSF group and 20 of 25 (80%) evaluable control patients had complete remission after phase I of induction therapy. We conclude that G-CSF can be safely administered as an adjunct to induction therapy of ALL and is clinically beneficial by ameliorating neutropenia and reducing infectious complications. PMID- 9226160 TI - Inhibition of immature erythroid progenitor cell proliferation by macrophage inflammatory protein-1alpha by interacting mainly with a C-C chemokine receptor, CCR1. AB - Several lines of evidence indicate that macrophage inflammatory protein-1alpha (MIP-1alpha) modulates the proliferation of hematopoietic progenitor cells, depending on their maturational stages. To clarify the mechanisms for the modulation of hematopoiesis by this chemokine, we examined the expression of a receptor for MIP-1alpha, CCR1, on bone marrow cells of normal individuals using a specific antibody and explored the effects of MIP-1alpha on in vitro erythropoiesis driven by stem cell factor (SCF) and erythropoietin (Epo). CCR1 was expressed on glycophorin A-positive erythroblasts in addition to lymphocytes and granulocytes. CCR1+ cells, isolated from bone marrow mononuclear cells (BMMNCs) using a cell sorter, comprised virtually all erythroid progenitor cells in the BMMNCs. Moreover, MIP-1alpha inhibited, in a dose-dependent manner, colony formation by burst-forming unit-erythroid (BFU-E), but not by colony forming unit erythroid (CFU-E), in a methylcellulose culture of purified human CD34+ bone marrow cells. Although reverse-transcription polymerase chain reaction (RT-PCR) showed the presence of CCR1, CCR4, and CCR5 transcripts in CD34+ cells in BM, anti-CCR1 antibodies significantly abrogated the inhibitory effects of MIP-1alpha on BFU-E formation both in a methylcellulose culture and in a single cell proliferation assay of purified CD34+ cells. Although the contribution of CCR4 or CCR5 cannot be completely excluded, these results suggest that MIP-1alpha mediated suppression of the proliferation of immature, but not mature erythroid progenitor cells, is largely mediated by CCR1 expressed on these progenitor cells. PMID- 9226159 TI - Jak1 plays an essential role for receptor phosphorylation and Stat activation in response to granulocyte colony-stimulating factor. AB - The proliferation and differentiation of neutrophils is regulated by granulocyte specific colony-stimulating factor (G-CSF). G-CSF uses a receptor of the cytokine receptor superfamily and, in common with all members of the family, induces the tyrosine phosphorylation and activation of members of the Janus protein tyrosine kinase (Jak) family. In both myeloid cells and a human fibrosarcoma cell line expressing the G-CSF receptor, G-CSF induces the tyrosine phosphorylation and activation of Jak1, Jak2, and Tyk2. In addition, G-CSF induces the tyrosine phosphorylation of the receptor and members of the signal transducers and activators of transcription (Stat) family, including Stat3, as well as Stat1 and Stat5, depending on the cells involved. Using mutant cell lines lacking various Jaks, we show here that Jak1 is critical for G-CSF-mediated Stat activation, whereas Jak2 or Tyk2 are either not required or play redundant or ancillary roles. In the absence of Jak1, G-CSF induces activation of Jak2 and Tyk2, but fails to induce receptor tyrosine phosphorylation and induces dramatically reduced levels of Stat activation. A kinase-inactive Jak2, when overexpressed in cells lacking endogenous Jak2, can suppress Jak1 activation, receptor phosphorylation, and Stat activation, suggesting competition in the receptor complex either for Jak1 binding or substrates. Because the requirement for Jak1 is very similar to that previously shown for interleukin-6 signaling, the data support the concept that the G-CSF receptor and gp130 are both structurally and functionally similar. PMID- 9226161 TI - Thrombopoietin in patients with congenital thrombocytopenia and absent radii: elevated serum levels, normal receptor expression, but defective reactivity to thrombopoietin. AB - The pathophysiology of thrombocytopenia in the syndrome of thrombocytopenia with absent radii (TAR) is not yet understood. We examined thrombopoietin (TPO) serum levels and the in vitro reactivity of platelets to TPO in five patients affected with TAR syndrome. We found elevated TPO serum levels in all patients tested, excluding a TPO production defect as cause for thrombocytopenia in TAR syndrome. In addition, we found similar expression of the TPO receptor c-Mpl on the surface of platelets from TAR patients (5 of 5) and a similar molecular weight of the receptor as compared with healthy controls (4 of 4). Platelet response to adenosine diphosphate or thrombin receptor agonist peptide SFLLRN (TRAP) was normal in TAR patients. However, in contrast to results with healthy controls we could show absence of in vitro reactivity of platelets from TAR patients to recombinant TPO as measured by testing TPO synergism to adenine diphosphate and TRAP in platelet activation. TPO induced tyrosine phosphorylation of platelet proteins was completely absent (3 of 4) or markedly decreased (1 of 4). Our results indicate that defective megakaryocytopoiesis/thrombocytopoiesis in TAR syndrome is not caused by a defect in TPO production but a lack of response to TPO in the signal transduction pathway of c-Mpl. PMID- 9226162 TI - In vivo effects of Flt3/Flk2 ligand on mobilization of hematopoietic progenitors in primates and potent synergistic enhancement with granulocyte colony stimulating factor. AB - The Flt3 receptor is expressed in primitive hematopoietic cells and its ligand exerts proliferative effects on these cells in vitro in synergy with other cytokines. To expand on the functional properties of Flt3 ligand (FL) in vivo we treated nonhuman primates with FL and tested its ability to mobilize stem/progenitor cells when given alone or in combination with granulocyte colony stimulating factor (G-CSF) treatment. FL alone (200 microg/kg/day) mobilizes progenitors with slow kinetics and with a peak effect at the end of 2 weeks of treatment. The spectrum of mobilized progenitors includes myeloid, lymphoid, megakaryocytic, and osteoclastogenic but a low proportion of burst-forming unit (BFU)e. Bone marrow (BM) studies before and during the treatment suggested that proliferative effects in BM may have preceded effects on peripheral blood mobilization. To assess the synergy of FL with G-CSF in mobilization of progenitors we used two schemes: one in which G-CSF was used for the last 5 days of a 12-day treatment with FL; the other in which both cytokines were given concurrently for 5 days only (FL, 200 microg/kg; G-CSF, 100 microg/kg). Both schemes yielded much higher progenitor mobilization levels (peak levels of colony forming cells [CFSs] 41,000 to 95,000/mL blood) than observed with either FL (CFC 4,600 to 7,300/mL) or G-CSF (8,405 +/- 3,024/ mL) used alone at the same doses. Furthermore, there was a progressive and significant expansion of progenitors in vitro during 2 weeks in suspension cultures of mononuclear cells or of CD34+ cells only in the animal with the combined treatment. Likewise, substantial mobilization of osteoclastogenic progenitors was documented only with the combined treatment. Given the functional properties of FL, its synergistic mobilization with G-CSF, and its anticipated good tolerance (because of the absence of an effect on mast cell activation), a clinical use is projected for this cytokine in peripheral blood transplantation settings, as well as in experiments with ex vivo gene transfer. PMID- 9226163 TI - The roles of Bcl-X(L) and apopain in the control of erythropoiesis by erythropoietin. AB - Erythropoietin (EP) is required by late-stage erythroid progenitor cells to prevent apoptosis. Several lines of evidence suggest that it is this action of EP that regulates erythrocyte production in vivo. To study the control of apoptosis in mouse and human erythroblasts, the expression of members of the Bcl-2 family of proteins and the expression and activation of the apoptosis-linked cysteine protease Yama/CPP32/apopain were examined. These proteins have been implicated as regulators of apoptosis in several cell models. The Bcl-2 family members analyzed were Bcl-2, Bcl-X, Bax, Bad, Bak, A1, and Mcl-1. Bcl-X expression in proerythroblasts was highly EP-dependent. Bcl-X was strongly increased during the terminal differentiation stages of human and mouse erythroblasts, reaching maximum transcript and protein levels at the time of maximum hemoglobin synthesis. This increase in Bcl-X expression led to an apparent level of approximately 50 times the level in proerythroblasts. In contrast, neither mouse nor human erythroblasts expressed Bcl-2 transcript or protein. Bax and Bad proteins remained relatively constant throughout differentiation, but diminished near the time of enucleation. Bak protein was present in early erythroblasts, but diminished progressively during differentiation. EP deprivation in both mouse and human erythroblasts led to activation of the cysteine protease, apopain, as was indicated by cleavage of the proenzyme into its proteolytically active fragments. Apopain activation was detectable within 2 hours of EP deprivation in mouse erythroblasts. These findings suggest an important role for Bcl-X in late erythroid differentiation and for apopain in apoptosis of erythroblasts caused by deprivation of EP. PMID- 9226164 TI - Differential maintenance of primitive human SCID-repopulating cells, clonogenic progenitors, and long-term culture-initiating cells after incubation on human bone marrow stromal cells. AB - Many experimental and clinical protocols are being developed that involve ex vivo culture of human hematopoietic cells on stroma or in the presence of cytokines. However, the effect of these manipulations on primitive hematopoietic cells is not known. Our severe combined immune-deficient mouse (SCID)-repopulating cell (SRC) assay detects primitive human hematopoietic cells based on their ability to repopulate the bone marrow (BM) of immune-deficient non-obese diabetic/SCID (NOD/SCID) mice. We have examined here the maintenance of SRC, colony-forming cells (CFC), and long-term culture-initiating cells (LTC-IC) during coculture of adult human BM or umbilical cord blood (CB) cells with allogeneic human stroma. Transplantation of cultured cells in equivalent doses as fresh cells resulted in lower levels of human cell engraftment after 1 and 2 weeks of culture for BM and CB, respectively. Similar results were obtained using CD34+-enriched CB cells. By limiting dilution analysis, the frequency of SRC in BM declined sixfold after 1 week of culture. In contrast to the loss of SRC as measured by reduced repopulating capacity, the transplanted inocula of cultured cells frequently contained equal or higher numbers of CFC and LTC-IC compared with the inocula of fresh cells. The differential maintenance of CFC/LTC-IC and SRC suggests that SRC are biologically distinct from the majority of these in vitro progenitors. This report demonstrates the importance of the SRC assay in the development of ex vivo conditions that will allow maintenance of primitive human hematopoietic cells with repopulating capacity. PMID- 9226165 TI - Identification of increased protein tyrosine phosphatase activity in polycythemia vera erythroid progenitor cells. AB - Polycythemia vera (PV) is a clonal hematologic disease characterized by hyperplasia of the three major bone marrow lineages. PV erythroid progenitor cells display hypersensitivity to several growth factors, which might be caused by an abnormality of tyrosine phosphorylation. In the present study, we have investigated protein tyrosine phosphatase (PTP) activity in highly purified erythroid progenitor cells and found that the total PTP activity in the PV cells was twofold to threefold higher than that in normal cells. Protein separation on anion-exchange and gel-filtration columns showed that the increased activity was due to a major PTP eluted at approximately 170 kD. This enzyme was sensitive to PTP inhibitors and it did not cross-react with antibodies to SHP-1, SHP-2, or CD45. Subcellular fractionation showed that the PTP localized with the membrane fraction, where its activity was increased by threefold in PV erythroid progenitors when compared with normal cells. As the erythroid progenitors progressively matured, activity of the PTP declined rapidly in the normal cells but at a much slower rate in the PV cells. These studies suggest that a potentially novel membrane or membrane-associated PTP, representing a major PTP activity, may have an important role in proliferation and/or survival of human erythroid progenitors and that its hyperactivation in PV erythroid progenitors might be responsible for the increased erythropoiesis in PV patients. PMID- 9226166 TI - Orderly process of sequential cytokine stimulation is required for activation and maximal proliferation of primitive human bone marrow CD34+ hematopoietic progenitor cells residing in G0. AB - Bone marrow (BM) CD34+ cells residing in the G0 phase of cell cycle may be the most suited candidates for the examination of cell cycle activation and proliferation of primitive hematopoietic progenitor cells (HPCs). We designed a double simultaneous labeling technique using both DNA and RNA staining with Hoechst 33342 and Pyronin Y, respectively, to isolate CD34+ cells residing in G0(G0CD34+). Using long-term BM cultures and limiting dilution analysis, G0CD34+ cells were found to be enriched for primitive HPCs. In vitro proliferation of G0CD34+ cells in response to sequential cytokine stimulation was examined in a two-step assay. In the first step, cells received a primary stimulation consisting of either stem cell factor (SCF), Flt3-ligand (FL), interleukin-3 (IL 3), or IL-6 for 7 days. In the second step, cells from each group were washed and split into four or more groups, each of which was cultured again for another week with one of the four primary cytokines individually, or in combination. Tracking of progeny cells was accomplished by staining cells with PKH2 on day 0 and with PKH26 on day 7. Overall examination of proliferation patterns over 2 weeks showed that cells could progress into four phases of proliferation. Phase I contained cytokine nonresponsive cells that failed to proliferate. Phase II contained cells dividing up to three times within the first 7 days. Phases III and IV consisted of cells dividing up to five divisions and greater than six divisions, respectively, by the end of the 14-day period. Regardless of the cytokine used for primary stimulation, G0CD34+ cells moved only to phase II by day 7, whereas a substantial percentage of cells incubated with SCF or FL remained in phase I. Cells cultured in SCF or FL for the entire 14-day period did not progress beyond phase III but proliferated into phase IV (with <20% of cells remaining in phases I and II) if IL-3, but not IL-6, was substituted for either cytokine on day 7. G0CD34+ cells incubated with IL-3 for 14 days proliferated the most and progressed into phase IV; however, when SCF was substituted on day 7, cells failed to proliferate into phase IV. Most intriguing was a group of cells, many of which were CD34+, detected in cultures initially stimulated with IL-3, which remained as a distinct population, mostly in G0/G1, unable to progress out of phase II regardless of the nature of the second stimulus received on day 7. A small percentage of these cells expressed cyclin E, suggesting that their proliferation arrest may have been mediated by a cyclin-related disruption in cell cycle. These results suggest that a programmed response to sequential cytokine stimulation may be part of a control mechanism required for maintenance of proliferation of primitive HPCs and that unscheduled stimulation of CD34+ cells residing in G0 may result in disruption of cell-cycle regulation. PMID- 9226167 TI - A three amino acid deletion in glycoprotein IIIa is responsible for type I Glanzmann's thrombasthenia: importance of residues Ile325Pro326Gly327 for beta3 integrin subunit association. AB - Glanzmann's thrombasthenia (GT) is a recessive autosomal bleeding disorder characterized by abnormal platelet aggregation due to a qualitative or quantitative defect of the glycoprotein (GP) IIb-IIIa complex (integrin alphaIIb beta3). We describe a new mutation in the GPIIIa gene responsible for type I GT in a consanguineous Algerian family. A discordance between phenotyping and genotyping of the GPIIIa-related HPA-1 platelet alloantigen system in three family members heterozygous for the disease suggested a genetic defect in the GPIIIa gene and a normal GPIIb gene. Sequence analysis of amplified genomic DNA fragments showed a 6-bp deletion in exon 7 of the GPIIIa gene resulting in the amino acid deletion/substitution (Ile325pro326Gly327 --> Met) and creating a new BspHI restriction site. Expression of the mutated integrin beta3 subunit cDNA in Chinese hamster ovary cells showed that the cDNA gene was transcribed into a full length beta3 protein with an apparent molecular weight identical to wild-type beta3 and accumulated as a single-chain molecule in the cell cytoplasm. The absence of heterodimeric complex formation of the mutant beta3 protein with endogenous alpha v was shown by immunoprecipitation experiments, intracellular immunofluorescent labeling, and a semiquantitative enzyme-linked immunosorbent assay using the alpha vbeta3 complex-specific monoclonal antibodies LM609 and 23C6. Substitution of the methionine residue by a proline, present at position 326 of wild-type beta3, did not restore the ability of the recombinant mutant beta3 protein to associate with alpha v , suggesting that the Ile-Pro-Gly motif is located in a beta3 domain important for integrin subunit interaction. The association of a BspHI restriction site with this newly identified mutation has allowed allele-specific restriction analysis of Algerian GT individuals and the identification of two new unrelated type I patients exhibiting the same mutation, suggesting that the described mutation might be significant in this population and that BspHI restriction analysis will provide a useful screening assay for antenatal diagnosis and genetic counselling. PMID- 9226168 TI - Binding of phosphorylated Sp1 protein to tandem Sp1 binding sites regulates alpha2 integrin gene core promoter activity. AB - The alpha2beta1 integrin, a collagen/laminin receptor, is expressed by a variety of cell types, including epithelial cells, mesenchymal cells, and hematopoietic cells. To understand the molecular mechanisms that regulate expression of the alpha2beta1, integrin in cells with megakaryocytic differentiation, we characterized the 5' flanking region of the alpha2 integrin gene and identified three distinct regulatory regions, including a core promoter, a silencer, and megakaryocyte enhancers in the distal 5' flank (Zutter et al, Blood 96:3006, 1995 and Zutter et al, J Biol Chem 269:463, 1994). We now focus on the core promoter of the alpha2 integrin gene located between bp -30 and -92 that is required for transcriptional activity of the alpha2 integrin gene. Sequence analysis identified two Sp1 consensus sites and a potential AP2 site. Gel retardation assays showed that nuclear proteins from uninduced K562 cells and K562 cells induced to become megakaryocytic bound specifically to the core promoter region (bp -30 to bp -92) producing two DNA-protein complexes. In addition, nuclear extracts from cells induced along the megakaryocyte lineage produced a selective increase in the slower migrating complex. Site-directed mutagenesis of the 5', the 3', or both Sp1 binding sites suggested that both Sp1 binding sites are required for full promoter activity and for DNA-protein complex formation. DNA footprinting also showed specific protection of the 5' Sp1 site by nuclear extracts from uninduced K562 cells and protection of both the 5' and the 3' Sp1 sites by nuclear extracts from induced K562 cells. Sp1 protein-DNA complex formation was dependent on Sp1 phosphorylation. The faster migrating DNA-protein complex was enhanced by dephosphorylation; the slower migrating DNA-protein complex was diminished or lost. PMID- 9226169 TI - High molecular weight kininogen peptides inhibit the formation of kallikrein on endothelial cell surfaces and subsequent urokinase-dependent plasmin formation. AB - A sequence of 31 amino acids (S565-K595) in domain 6 of the light chain of high molecular weight kininogen (HK) has previously been shown to be responsible for the binding of plasma prekallikrein (PK) or kallikrein. To find effective peptides that might block binding between HK and PK on cell surfaces, a new series of synthetic peptides has now been prepared that incorporates portions of this binding domain sequence. For mapping the minimal sequence within HK, these new peptides were tested for their ability to compete with HK for binding PK in a cell-free system and on human umbilical vein endothelial cells (HUVEC). In the former, at pH 7.4, the kds for binding between kallikrein and either D567-K595, S565-P594, D567-S593, or D567-T591 were all similar to that for the binding of S565-K595 (0.2 to 0.4 micromol/L), but those for the binding of D568-K595, W569 K595, and D567-P589 were an order of magnitude greater (kd = 2 to 5 micromol/L). D567-S586, the shortest chain length of the N- and C-terminal truncation sequences tested, does not effectively compete with kininogen for kallikrein binding (kd = 100 micromol/L). These results imply that D567-T591, a 25-residue peptide (HK25c), contains sufficient structural information for binding kallikrein in solution. D567-T591 also is the minimum structural sequence to block binding of kallikrein to HUVEC-bound HK (IC50 = 50 nmol/L) and to inhibit PK activation to kallikrein on the cell surface (IC50 = 80 nmol/ L). In addition, D567-T591 also inhibits the generation of kallikrein-activated urokinase, which activates plasminogen to plasmin (IC50 = 100 nmol/L). Thus, HK-derived peptides may be useful compounds for modulating excessive fibrinolysis and hypotension in sepsis and multiple trauma. PMID- 9226170 TI - Expression and functional characterization of an abnormal platelet membrane glycoprotein Ib alpha (Met239 --> Val) reported in patients with platelet-type von Willebrand disease. AB - Platelet-type von Willebrand disease (vWD) is a congenital bleeding disorder characterized by heightened ristocetin-induced platelet aggregation caused by abnormally high affinity between the platelet membrane glycoprotein (GP) Ib/IX complex and von Willebrand factor (vWF). Two distinct point mutations, Gly233 to Val and Met239 to Val, have been reported in GPIb alpha. We have constructed a recombinant GPIb alpha fragment containing the latter mutation, Met239 to Val (M239V) and characterized the mutant molecule using two methods, ie, interaction between soluble vWF and immobilized M239V and inhibition of platelet aggregation by purified soluble M239V. Spontaneous binding (ie, binding without any inducers) was observed between 125I-vWF and immobilized M239V but not between 125I-vWF and immobilized wild-type (WT) GPIb alpha. The addition of low concentrations of ristocetin (0.2 mg/mL) induced specific 125I-vWF binding to immobilized M239V, but not to WT GPIb alpha. At high concentrations of ristocetin (1.2 mg/mL), both WT GPIb alpha and M239V specifically bound to 125I-vWF. Thus, M239V reproduced the unique functional abnormality of the GPIb/IX complex in platelet-type vWD. Moreover, the purified soluble M239V inhibited platelet aggregation induced by low concentration of ristocetin (0.3 mg/mL) in platelet-rich plasma from a patient having Met239 to Val mutation, whereas purified WT did not. These results provide direct evidences that the reported point mutation is the responsible molecular basis of this disorder. PMID- 9226171 TI - Regulation of tie receptor expression on human endothelial cells by protein kinase C-mediated release of soluble tie. AB - The expression and activity of receptor tyrosine kinases (RTK) at the cell surface can be modulated by several different pathways including the proteolytic release of the extracellular domain as a soluble receptor. We investigated the regulation of tie receptor expression, an orphan RTK restricted to cells of hematopoietic and endothelial lineages, on primary human endothelial cells and a stably transfected Chinese hamster ovary (CHO) cell line. Tie was expressed in cells as a doublet of 135 and 125 kD; the 135-kD band represented mature cell surface receptor containing sialic acid and N-linked oligosaccharide residues, whereas the 125-kD band represented an intracellular pool of immature receptor. Phorbol 12-myristate 13-acetate (PMA) had dramatic effects on tie expression at the cell surface. Within 15 minutes of PMA treatment, the 135-kD band disappeared from the cell surface and was accompanied by the appearance of a 100-kD band in cell supernatants. The 100-kD band continued to accumulate in the media throughout the duration of PMA treatment during which mature tie receptor was undetectable on the cell surface by fluorescence-activated cell sorting (FACS) or in cell lysates by immunoblot analysis. Using specific antibodies, this 100-kD species was shown to be a soluble form of the tie receptor containing the extracellular domain. PMA-dependent release of soluble tie was mediated through the activation of protein kinase C (PKC); soluble tie was not released in the presence of PKC inhibitors, an inactive PMA analog, or following the downregulation of PKC through chronic PMA treatment. These results indicate that tie receptor expression on endothelial cells is regulated by the release of a soluble extracellular fragment following activation of PKC. Parallel pathways regulating c-kit, tumor necrosis factor (TNF), and colony-stimulating factor (CSF) receptor expression suggest that the release of extracellular receptor fragments represents an alternative mechanism through which cells modulate responses to growth factors and cytokines. PMID- 9226173 TI - Antibodies to HLA class I alpha1 domain trigger apoptosis of CD40-activated human B lymphocytes. AB - We analyzed herein whether antibodies to HLA class I alpha1 domain, which trigger apoptosis of activated T cells, may also control the growth/survival of human B lymphocytes. Addition of monoclonal antibody (MoAb) 90 (mouse IgG1) or YTH862 (rat IgG2b) was found to strongly inhibit the proliferation of CD40-activated total tonsil B cells as well as that of purified naive, germinal center, and memory B-cell subsets. This inhibitory effect was not prevented by addition of B cell tropic factors, such as interleukin-2 (IL-2), IL-4, and IL-10, and was a result of induced B-cell apoptosis as shown by using a TUNEL assay and DNA electrophoresis. In contrast, engagement of another epitope of the alpha1 domain, as well as that of the alpha2 and alpha3 domains by specific anti-HLA class I MoAbs, failed to inhibit DNA synthesis and to induce apoptosis of CD40-activated B cells. As recently reported for acquisition of sensitivity to Fas (APO-1/CD95) dependent apoptosis, susceptibility to MoAb90-and YTH862-induced death was restricted to CD40-activated B cells, because resting and anti-IgM-activated B cells did not undergo apoptosis after HLA class I engagement. Moreover, ligation of the B-cell receptor protected CD40-activated B cells from both HLA class I- and Fas-mediated growth inhibition and apoptosis. Taken together, these results show that engagement of the alpha1 domain of HLA class I induces apoptotic cell death of CD40-activated, but not of antigen-activated B cells, and would, therefore, suggest a possible role for HLA class I molecules in the control of B cell homeostasis. PMID- 9226172 TI - Signaling pathways regulating CD44-dependent cytolysis in natural killer cells. AB - CD44 is a cytotoxic triggering molecule on activated, but not fresh natural killer (NK) cells. In the current study, metabolic pathways used in CD44-directed lysis (CD44DL) were examined using activated human NK cells as effectors. We found that CD44 expressed by activated NK cells was indistinguishable in isoform and molecular weight from CD44 on unactivated cells. However, de novo protein expression was required for the induction of CD44DL, suggesting that activated NK cells contain proteins not present in fresh NK cells that couple CD44 to the lytic machinery. Concanimycin A, a selective inhibitor of perforin-based cytolysis, totally blocked CD44DL, natural cytotoxicity, and antibody-dependent cell-mediated cytolysis (ADCC). Moreover, studies in which kinase inhibitors were added during the effector phase of lysis indicated that protein-tyrosine and ser/thr kinases were required for all three cytolytic activities and that protein kinase C played a nonessential role in lysis. By contrast, wortmannin totally inhibited CD44DL, but failed to block natural cytotoxicity and only partially blocked ADCC, suggesting that phosphatidylinositol 3-kinase (PI 3-kinase) is required at an early, receptor-specific stage of CD44DL. Finally, cytochalasin B enhanced CD44DL, but not ADCC, indicating that CD44DL is modulated by actin polymerization. Taken together, our data suggest that CD44 in NK cells interacts with proteins induced during interleukin-2 activation in a triggering pathway that induces perforin release, requires PI 3-kinase, and is modulated by the cytoskeleton. PMID- 9226174 TI - Ig D(H) gene segment transcription and rearrangement before surface expression of the pan-B-cell marker CD19 in normal human bone marrow. AB - The onset of IgH transcription and rearrangement is a defining characteristic of the progenitor population in which B-lineage commitment occurs. These features were used to better define the earliest stage of B-cell commitment in humans and to determine if these stages differ as a function of human ontogeny. Fetal and adult bone marrow mononuclear cells were sorted into B-lineage subpopulations on the basis of surface expression of the stem cell marker CD34, the pan-B-cell marker CD19, and IgM and analyzed for transcription and rearrangement of the IgH locus. The locus was found to be transcriptionally active before surface expression of CD19, as indicated by the presence of germline I mu, C mu, and D(H)Q52 transcripts in the CD34+ CD19- subpopulation. Transcripts from IgH alleles that had undergone DJC mu rearrangements were also detected in the CD34+ CD19- subpopulation. Within this subpopulation, low levels of DXP-containing DJC mu transcripts were detected in both fetal and adult cells. Although D(H)Q52 DJC mu transcripts were abundant in fetal CD34+ CD19- cells, they were not detected in cells of the same phenotype derived from adult bone marrow. In both fetus and adult, V(H)3-and V(H)6-containing VDJC mu transcripts were detected only in the CD19+ subpopulations. These data indicate that transcription of D(H)Q52-J(H) and DXP-J(H) rearrangements differs during fetal and adult B lymphopoiesis. Moreover, in both fetus and adult, transcription of unrearranged components of the IgH locus and DJ rearrangements can proceed before the surface expression of CD19. PMID- 9226175 TI - Modulation of Bcl-2 protein by CD4 cross-linking: a possible mechanism for lymphocyte apoptosis in human immunodeficiency virus infection and for rescue of apoptosis by interleukin-2. AB - We have previously demonstrated that CD4 cross-linking (CD4XL) results in apoptosis of CD4+ T cells and augmentation of Fas antigen (CD95, APO-1) expression in CD4+ and CD8+ T cells. Here we demonstrate that CD4XL mediated by both, anti-CD4 monoclonal antibody (MoAb) or human immunodeficiency virus (HIV) envelope protein gp120 reduces the expression of the proto-oncogene Bcl-2 in CD4+ T cells, but not in CD8+ T cells, concurrently with the induction of CD4+ T cell apoptosis. Additionally, the Bcl-2dim population expressed high levels of Fas antigen. Bax, an antagonist of Bcl-2, was brightly expressed even in the Bcl-2dim population. Addition of interleukin (IL)-2 rescued CD4+ T cells from CD4XL induced Bcl-2 downmodulation and apoptosis induction. These results support the hypothesis that CD4 ligation by HIV-1 envelope protein in vivo in HIV-infected patients selectively reduces Bcl-2 protein in CD4+ T lymphocytes, thereby facilitating Fas/Fas-ligand triggered apoptosis; furthermore the findings reported expand the rationale for use of IL-2 in HIV disease. PMID- 9226176 TI - The development of a model for the homing of multiple myeloma cells to human bone marrow. AB - Prior in vitro studies have suggested a role of adhesion molecules, bone marrow stromal cells (BMSCs), and cytokines in the regulation of human multiple myeloma (MM) cell growth and survival. Although in vivo models have been developed in severe combined immunodeficient (SCID) mice that support the growth of human MM within the murine BM microenvironment, these xenograft models do not permit a study of the role of adhesion proteins in human MM cell-human BMSC interactions. We therefore established an in vivo model of human MM using SCID mice implanted with bilateral human fetal bone grafts (SCID-hu mice). For the initial tumor innoculum, human MM derived cell lines (1 x 10(4) or 5 x 10(4) ARH-77, OCI-My5, U 266, or RPMI-8226 cells) were injected directly into the BM cavity of the left bone implants in irradiated SCID-hu mice. MM cells engrafted and proliferated in the left human fetal bone implants within SCID-hu mice as early as 4 weeks after injection of as few as 1 x 10(4) MM cells. To determine whether homing of tumor cells occurred, animals were observed for up to 12 weeks after injection and killed to examine for tumor in the right bone implants. Of great interest, metastases to the right bone implants were observed at 12 weeks after the injection of 5 x 10(4) MM cells, without spread of human MM cells to murine BM. Human MM cells were identified on the basis of characteristic histology and monoclonal human Ig. Importantly, monoclonal human Ig and human interleukin-6 (IL 6), but not human IL-1beta or tumor necrosis factor-alpha, were detectable in sera of SCID-hu mice injected with MM cells. In addition, specific monoclonal Ig light chain deposition was evident within renal tubules. This in vivo model of human MM provides for the first time a means for identifying adhesion molecules that are responsible for specific homing of human MM cells to the human, as opposed to murine, BM microenvironment. Moreover, induction of human IL-6 suggests the possibility that regulation of MM cell growth by this cytokine might also be investigated using this in vivo model. PMID- 9226177 TI - Lymphomas in patients with Sjogren's syndrome are marginal zone B-cell neoplasms, arise in diverse extranodal and nodal sites, and are not associated with viruses. AB - The occurrence of non-Hodgkin's lymphoma (NHL) is the most serious complication of Sjogren's syndrome (SS). We performed a study of 16 NHLs occurring in patients with an underlying SS. These lymphomas arose not only in salivary glands (7 cases) but also in other mucosal extranodal sites (the stomach [4 cases], the lung [3 cases], the skin [3 cases], the buccal mucosa [1 case], the thymus [1 case]) and in nodal sites (8 cases). Low-grade marginal zone lymphomas (MZL) were diagnosed in 12 of the 16 patients, 9 of mucosa-associated lymphoid tissues (MALT) type in mucosal sites and 3 exclusively nodal. The 4 other patients presented with a high-grade B-cell lymphoma that was probably a histological transformation of an underlying low-grade MZL at least in 3 of the cases involving skin, stomach, and parotid, respectively. A t(14;18) translocation was detected in 1 of 8 lymphomas tested. We detected serum anti-p53 antibodies in 2 of the 14 studied patients. p53 protein was detected in 1 of 11 lymphomas tested. LMP protein and Eber RNAs of Epstein-Barr virus (EBV) were not detected in the 16 NHL biopsies. Using polymerase chain reaction, EBV was never detected except in 1 of 4 parotid lymphomas. No human T-lymphotropic virus 1 or human herpes virus 8 DNAs were detected in NHL biopsies. None of the patients had hepatitis C virus infection found using serological methods. Chemotherapy was usually efficient. In conclusion, lymphomas occurring in patients with an underlying SS are in most cases MZL. These lymphomas are not associated with viruses known to be present in other types of lymphomas. Some of the translocations or mutations of oncogenes or antioncogenes described in other lymphomas are detected in SS-associated lymphomas. PMID- 9226179 TI - Cytokine expression and tumorigenicity of large granular lymphocytic leukemia cells from mice transgenic for the tax gene of human T-cell leukemia virus type I. AB - The human T-cell leukemia virus type I (HTLV-I) regulatory protein, Tax, has been speculated to play a major role in HTLV-I leukemogenesis. Indeed, several studies have suggested that upregulation of various cellular oncogenes and cytokines by Tax may explain the pathogenesis observed in HTLV-I-infected individuals, as well as several Tax-transgenic animal models. We report here the analysis of cytokine expression in a Tax-transgenic animal model with large granular lymphocytic (LGL) leukemia. Two different transgenic mice showed identical expression of interleukin-1alpha (IL-1alpha), IL-1beta, interferon gamma (IFNgamma), and granulocyte-macrophage colony-stimulating factor (GM-CSF) in peripheral tail tumors. Interestingly, LGL cell lines derived from these same tumors expressed high levels of both IFNgamma and GM-CSF, which correlated with the level of Tax expression. These same LGL cell lines also expressed high levels of lymphocyte function-associated antigen-1 (LFA-1) and intracellular adhesion molecule-1 (ICAM 1). Engraftment of these LGL cell lines into severe combined immunodeficient (SCID) mice led to the development of leukemia and lymphomas. Examination of these SCID mice showed that their pathology was nearly identical to that observed in the original Tax-transgenic mouse model. Both the Tax-transgenic and engrafted SCID mouse models allow for the analysis of cellular events that are required for tumor development associated with HTLV infection and suggest that Tax expression may be responsible for the upregulation of certain cytokines and adhesion molecules that affect the infiltrating capabilities of HTLV-I-infected cells. PMID- 9226178 TI - Characterization of overt B-cell lymphomas in patients with hepatitis C virus infection. AB - A pathogenetic role of the hepatitis C virus (HCV) has been hypothesized for a subset of B-cell non-Hodgkin's lymphomas (NHLs). However, the preliminary characterization of B-cell NHLs in HCV-infected individuals has been poorly addressed. In the present study, we report detailed information on 35 consecutive patients with overt B-cell NHL of recent onset and HCV infection; all patients referred to a single oncological center in Northeast Italy. Histopathologic evaluation was performed by a single reference hemopathologist, and the link with the two relevant autoimmune diseases predisposing to B-cell NHL and in which HCV has been implied, ie, "essential" mixed cryoglobulinemia (EMC) and Sjogren's syndrome, was investigated. Control groups included 122 consecutive HCV-negative patients with B-cell NHL and 464 consecutive histopathologic cases of B-cell NHL referred to the same center, as well as 127 consecutive patients with HCV infection and without lymphoma referred to a different center in the same geographical area. B-cell NHLs in HCV-infected patients frequently presented at onset (1) an extranodal localization with peculiar target organs of HCV infection (ie, the liver and major salivary glands) being significantly overrepresented; (2) a diffuse large cell histotype without any prior history of low-grade B-cell malignancy or bone marrow involvement; and (3) a weak association with a full blown predisposing autoimmune disease, although serum autoimmune features were common and cryoglobulins were always present. Therefore, the HCV-related B-cell NHLs in this oncological series presented distinctive features compared with B cell NHLs in HCV-negative patients, and they differed from bone marrow low-grade NHLs frequently diagnosed in HCV-positive patients with EMC. Such novel information may be relevant for future research aimed at clarifying the possible link between HCV infection, autoimmunity, nonmalignant B-cell lymphoproliferation, and overt B-cell malignancy. PMID- 9226180 TI - A specific stimulator of granulocyte colony-stimulating factor accelerates recovery from cyclophosphamide-induced neutropenia in the mouse. AB - We have identified a small molecular weight compound, SCH 14988, which specifically stimulates in vitro granulocyte-colony stimulating factor (G-CSF) production from activated human peripheral blood mononuclear cells and monocytes but not other cytokines or CSFs with hematoregulatory activity. In vivo administration of SCH 14988 to mice rendered neutropenic by cyclophosphamide treatment resulted in the accelerated recovery of the peripheral neutrophil compartment. This activity correlated with increased in vivo G-CSF levels and stimulation of marrow granulopoiesis, and was comparable to that of exogenously administered recombinant human G-CSF. No alterations to other leukocyte populations in peripheral blood, spleen, or the peritoneal cavity were observed. These findings suggest that SCH 14988 may be clinically useful to enhance neutrophil granulopoiesis, as well as to study the mechanisms involved in G-CSF gene regulation. PMID- 9226181 TI - Neutrophil secondary-granule deficiency as a hallmark of all-trans retinoic acid induced differentiation of acute promyelocytic leukemia cells. AB - Acute promyelocytic leukemia (APL) is a neoplasm with the unique chromosomal translocation t(15;17), which involves the retinoic acid receptor alpha gene. All trans retinoic acid (ATRA) has been used for APL patients as a potent therapeutic agent to induce differentiation of leukemia cells. Although polymorphonuclear leukocytes (PMNs) appearing in the blood and bone marrow during ATRA treatment often possess Auer rods, indicating their neoplastic origin, other morphological abnormalities of PMNs have not been elucidated. We studied the morphological changes of APL cells during ATRA treatment at the ultrastructural level. Although most aberrant primary granules, including Auer rods, became morphologically normal in response to ATRA therapy and the nuclei showed chromatin condensation and lobulation, resulting in the emergence of PMNs, the lobulated nuclei often had nuclear filamentous connections and/or nuclear blebs, indicating some pathological process. Furthermore, PMNs, particularly early in ATRA treatment, lacked neutrophil secondary granules as did the PMNs appearing in a culture of APL cells incubated with ATRA, findings consistent with previously reported data that acute myeloid leukemia cell lines do not produce secondary granule proteins even after induction of differentiation towards mature neutrophils. The present data indicate that ATRA is incapable of inducing complete morphological maturation of APL cells and that secondary-granule deficiency may be a hallmark of aberrantly differentiated leukemic cells. PMID- 9226182 TI - Hereditary hyperferritinemia-cataract syndrome: relationship between phenotypes and specific mutations in the iron-responsive element of ferritin light-chain mRNA. AB - Recent reports have described families in whom a combination of elevated serum ferritin not related to iron overload and congenital nuclear cataract is transmitted as an autosomal dominant trait. We have studied the molecular pathogenesis of hyperferritinemia in two families showing different phenotypic expression of this new genetic disorder. Serum ferritin levels ranged from 950 to 1,890 microg/L in affected individuals from family 1, and from 366 to 635 microg/L in those from family 2. Cataract was clinically manifested in family 1 and asymptomatic in family 2. By using monoclonal antibodies specific for the H and L ferritin subunits, serum ferritin was found to be essentially L type in both normal and affected individuals. The latter also showed normal amounts of H type ferritin in circulating mononuclear cells; on the contrary, L-type ferritin contents were 13 times normal in family 1 and five times normal in family 2 on average. Serum ferritin was glycosylated in both normal and affected individuals. There was a close relationship between mononuclear cell L-type ferritin content and serum ferritin concentration (r = 0.95, P < .00001), suggesting that the excess production of ferritin in cells was directly responsible for the hyperferritinemia. The dysregulated L-subunit synthesis was found to result from different point mutations in a noncoding sequence of genomic L-subunit DNA, which behaves as an mRNA cis-acting element known as iron regulatory element (IRE). Affected individuals from family 1 were heterozygous for a point mutation (a single G to A change) in the highly conserved, three-nucleotide motif forming the IRE bulge. Affected members from family 2 were heterozygous for a double point mutation in the IRE lower stem. Using a gel retardation assay, the observed molecular lesions were shown to variably reduce the IRE affinity for an iron regulatory protein (IRP), which inhibits ferritin mRNA translation. The direct relationship between the degree of hyperferritinemia and severity of cataract suggests that this latter is the consequence of excessive ferritin production within the lens fibers. These findings provide strong evidence that serum ferritin is a byproduct of intracellular ferritin synthesis and that the L subunit gene on chromosome 19 is the source of glycosylated serum ferritin. From a practical standpoint, this new genetic disorder should be taken into account by clinicians when facing a high serum ferritin in an apparently healthy person. PMID- 9226183 TI - Pyridoxine refractory X-linked sideroblastic anemia caused by a point mutation in the erythroid 5-aminolevulinate synthase gene. AB - To elucidate how pyridoxine-refractory X-linked sideroblastic anemia (XLSA) develops, we analyzed the erythroid-specific 5-aminolevulinate synthase (ALAS-E) gene of a patient with the anemia. The activity and amount of the enzyme in bone marrow cells of the patient were found to be approximately 5% of the normal control. We identified a point mutation, which introduces an amino acid substitution from Asp 190 to Val. In transient transfection analyses using quail fibroblasts, accumulation of aberrantly processed proteins, the sizes of which were larger than that of mature ALAS-E, was found in mitochondria. The proteins were reproducibly detected in assays combining in vitro transcription/translation of ALAS-E precursor and import of the precursor into isolated mouse mitochondria. These results suggest that the mutation causing pyridoxine-refractory XLSA affects the processing of the ALAS-E precursor, thus provoking instability of the ALAS-E protein. PMID- 9226184 TI - Utilization of intracellular ferritin iron for hemoglobin synthesis in developing human erythroid precursors. AB - Ferritin (Ft) plays an important role in cellular iron metabolism. It can store substantial amounts of iron in a nontoxic soluble form. However, its ability to donate iron for cellular needs, in particular for hemoglobin (Hb) synthesis in human erythroid cells, is still controversial. We studied the role of intracellular Ft-iron in Hb synthesis and the involvement of lysosomal proteolysis in iron release from Ft. Ft-iron release and its subsequent incorporation into heme was investigated in normal human erythroid precursors developing in culture. Dual staining flow cytometry with antibody (Ab)-specific for Ft and for Hb showed a decrease in cellular Ft content in erythroid cells during their maturation. Cellular Ft-iron participation in heme synthesis was studied by labeling cells with 59Fe. Cells were incubated with 59Fe-labeled human diferric transferrin (Tf), then chased, and intracellular radioiron distribution between Ft and Hb was determined on subsequent days by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and/or Ft immunoprecipitation and heme extraction. On day 6, most of the 59Fe accumulated in Ft. Thereafter, a progressive decrease of radioiron in Ft and a corresponding increase of the label in Hb was observed. Inhibition of heme synthesis with succinylacetone caused radioiron to remain in Ft and prevented its redistribution. Addition of unlabeled diferric Tf to the culture medium did not prevent radioiron from appearing in Hb. Chloroquine repression of lysosomal function prevented radio-iron redistribution between Ft and Hb. Inhibition of proteolysis by chymostatin and/or leupeptin led to Ft-protein accumulation in the cells and also prevented radioiron transfer from Ft to Hb. The results of the present study suggest that intracellular Ft donates iron for heme synthesis and that proteolytic Ft degradation in a lysosomal-like compartment is necessary for iron release and its transfer to heme. PMID- 9226185 TI - Wide variety of point mutations in the H gene of Bombay and para-Bombay individuals that inactivate H enzyme. AB - The H genes, encoding an alpha1,2fucosyltransferase, which defines blood groups with the H structure, of four Bombay and 13 para-Bombay Japanese individuals were analyzed for mutations. Four Bombay individuals were homologous for the same null H allele, which is inactivated by a single nonsense mutation at position 695 from G to A (G695A), resulting in termination of H gene translation. The allele inactivated by the G695A was designated h1. The other 13 para-Bombay individuals possessed a trace amount of H antigens on erythrocytes regardless of their secretor status. Sequence analysis of their H genes showed four additional inactivated H gene alleles, h2, h3, h4, and h5. The h2 allele possesed a single base deletion at position 990 G (990-del). The h3 and h4 alleles possessed a single missense mutation, T721C, which changes Tyr 241 to His, and G442T, which changes Asp148 to Tyr, respectively. The h5 allele possessed two missense mutations, T460C (Tyr154to His) and G1042A (Glu348to Lys). The h2, h3, h4, and h5 enzymes directed by these alleles were not fully inactivated by the deletion and the missense mutations expressing some residual enzyme activity resulting in synthesis of H antigen on erythrocytes. Thirteen para-Bombay individuals whose erythrocytes retained a trace amount of H antigen were determined to be heterozygous or homozygous for at least one of h2, h3, h4, or h5 alleles. This clarified that the levels (null to trace amount) of H antigen expression on erythrocytes of Bombay and para-Bombay individuals are determined solely by H enzyme activity. These mutations found in the Japanese H alleles differ from a nonsense mutation found in the Indonesian population. To determine the roles of the H, Se, and Le genes in the expression of H antigen in secretions and Lewis blood group antigen on erythrocytes, the Lewis and secretor genes were also examined in these Bombay and para-Bombay individuals. The Lewis blood group phenotype, Le(alpha- b+), was determined by the combinatorial activity of two fucosyltransferases, the Lewis enzyme and the secretor enzyme, and the secretor status was solely determined by the secretor enzyme activity, not by H enzyme activity. Bombay individuals were confirmed to be homozygous for the inactivated H and Se genes. As expected from the very low frequency of Bombay and para-Bombay individuals in the population, ie, approximately one in two or 300,000, the H gene mutations were found to be very variable, unlike the cases of the point mutations in the other glycosyltransferase genes; the ABO genes, the Lewis gene, and the secretor gene. PMID- 9226186 TI - Essential role of the thymus to reconstitute naive (CD45RA+) T-helper cells after human allogeneic bone marrow transplantation. AB - To contribute to the understanding of the role of the thymus in humans in the reconstitution of naive (CD45RA+) T cells after bone marrow transplantation (BMT), we compared T-cell regeneration in a unique situation, namely a thymectomized cancer patient (15 years old), with that of thymus-bearing patients after allogeneic BMT. These cases shared features of transplantation (total body irradiation, HLA-matched donors, and graft-versus-host disease prophylaxis with cyclosporine A) and all had an uncomplicated post-transplantation course. As shown by fluorescence-activated cell sorting analyses, the thymectomized host failed to reconstitute CD45RA+ T-helper cells even 24 months after BMT (11% CD45RA+ of CD4+ cells). In this patient, preferentially CD45RO+ cells contributed to the recovery of CD4+ cells (206 of 261/microL at 6 months and 463 of 558/microL at 24 months after BMT, CD45RA+ of CD4+ cells), whereas CD45RA+ cells remained low (<60/microL). In contrast, nine thymus-bearing hosts (5 children and 4 adults) examined between 6 and 24 months after BMT effectively reconstituted CD4+/CD45RA+ cells according to their normal age-related range (> or = 28% in adults and > or = 50% in children). Five of these were analyzed sequentially at 6 and 9 months after BMT. Within this period, CD45RA+ cells increasingly contributed to the recovery of CD4+ cells (median, +21%), even when total CD4+ cells decreased. With respect to T-cytotoxic/suppressor cells, the thymectomized host retained the capacity to recover CD45RA+ cells (137 of 333/microL at 6 months and 596 of 1,046/microL at 24 months after BMT, CD45RA+ of CD8+ cells), a proportion similar to that seen in thymus-bearing hosts. These findings suggest that a thymus-independent pathway exists to regenerate CD45RA+ T cytotoxic/suppressor cells, but residual thymus is essential to reconstitute naive (CD45RA+) T-helper cells after BMT in humans. PMID- 9226187 TI - Bone marrow transplantation for severe aplastic anemia: has outcome improved? AB - Bone marrow transplants for severe aplastic anemia were first performed in the 1970s. Transplant regimens, supportive care, and patient selection have changed substantially since then. Our objective was to determine the impact of these changes on transplant outcome. We studied 1,305 recipients of HLA-identical sibling transplants for aplastic anemia between 1976 and 1992, reported to the IBMTR by 179 centers. We compared survival of transplants performed in three intervals (1976 through 1980 [n = 186], 1981 through 1987 [n = 648], and 1988 through 1992 [n = 471]) using Cox proportional hazards regression. Five-year survival (+/-95% confidence interval) increased from 48% +/- 7% in the 1976-1980 cohort to 66% +/- 6% in the 1988-1992 cohort (P < .0001). Risks of graft-versus host disease (GVHD) and interstitial pneumonia decreased over time, but the risk of graft failure did not. Higher long-term survival resulted primarily from decreased mortality in the first 3 months posttransplantation. Late mortality risks were low and changed little over the intervals studied. In multivariate analysis, changes in transplantation strategies accounted for most but not all of the improved outcome. Use of cyclosporine to prevent GVHD was the most important factor. Changes in patient selection did not seem to explain improved survival. Survival after HLA-identical sibling bone marrow transplantations for aplastic anemia has improved since 1976. Changes in GVHD prophylaxis account for much of this improvement. Other changes may also operate. PMID- 9226188 TI - Cytokine-facilitated transduction leads to low-level engraftment in nonablated hosts. AB - Using a murine bone marrow transplantation model, we evaluated the long-term engraftment of retrovirally transduced bone marrow cells in nonmyeloablated hosts. Male bone marrow was stimulated in a cocktail of interleukin-3 (IL-3), IL 6, IL-11, and stem cell factor (SCF) for 48 hours, then cocultured on the retroviral producer line MDR18.1 for an additional 24 hours. Functional transduction of hematopoietic progenitors was detected in vitro by reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of multiple drug resistance 1 (MDR1) mRNA from high proliferative potential-colony forming cell (HPP-CFC) colonies. After retroviral transduction, male bone marrow cells were injected into nonablated female mice. Transplant recipients received three TAXOL (Bristol-Myers, Princeton, NJ) injections (10 mg/kg) over a 14-month period. Transplant recipient tissues were analyzed by Southern blot and fluorescence in situ hybridization for Y-chromosome-specific sequences and showed donor cell engraftment of approximately 9%. However, polymerase chain reaction amplification of DNAs from bone marrow, spleen, and peripheral blood showed no evidence of the transduced MDR1 gene. RT-PCR analysis of total bone marrow RNA showed that transcripts from the MDR1 gene were present in a fraction of the engrafted donor cells. These data show functional transfer of the MDR1 gene into nonmyeloablated murine hosts. However, the high rates of in vitro transduction into HPP-CFC, coupled with the low in vivo engraftment rate of donor cells containing the MDR1 gene, suggest that the majority of stem cells that incorporated the retroviral construct did not stably engraft in the host. Based on additional studies that indicate that ex vivo culture of bone marrow induces an engraftment defect concomitantly with progression of cells through S phase, we propose that the cell cycle transit required for proviral integration reduces or impairs the ability of transduced cells to stably engraft. PMID- 9226189 TI - Constitutive expression of a CD44 variant isoform on T cells facilitates regaining of immunocompetence in allogeneic bone marrow transplantation. AB - Constitutive expression of a rat CD44 variant isoform, rCD44v4-v7, on murine T cells accelerates immune responsiveness. Because prolonged immunodeficiency can be a major drawback in allogeneic bone marrow transplantation, we considered it of special interest to see whether repopulation of lethally irradiated syngeneic and allogeneic mice may be influenced by constitutive expression of the rCD44v4 v7 transgene. When lethally irradiated syngeneic and allogeneic mice were reconstituted with bone marrow cells (BMC) from rCD44v4-v7 transgenic (TG) or nontransgenic (NTG) mice, the former had a clear repopulation advantage: thymocytes expanded earlier after reconstitution and, as a consequence, higher numbers of lymphocytes were recovered from spleen and lymph nodes. Lymphocytes also displayed functional activity in advance to those from mice reconstituted with BMC from NTG mice. Most importantly, after the transfer of BMC from TG mice into an allogeneic host, the frequency of host-reactive T cells decreased rapidly. Apparently, this was due to accelerated induction of tolerance. Because these effects were counterregulated by an rCD44v6-specific antibody, it is likely that they could be attributed to the rCD44v4-v7 TG product. Thus, expression of a CD44 variant isoform at high levels facilitated reconstitution with allogeneic BMC by accelerated establishment of tolerance and the regaining of immunocompetence. PMID- 9226190 TI - Downregulation of increased CD95 (APO-1/Fas) ligand in T cells from human immunodeficiency virus-type 1-infected children after antiretroviral therapy. PMID- 9226191 TI - Severe proximal deep vein thrombosis in a Glanzmann thrombasthenia variant successfully treated with a low molecular weight heparin. PMID- 9226192 TI - Are activated cytotoxic T cells in Hodgkin's disease biopsies a poor prognostic marker? PMID- 9226193 TI - Valproic acid and augmentation of fetal hemoglobin in individuals with and without sickle cell disease. PMID- 9226194 TI - Prevalence of selective IgA deficiency in Spain: more than we thought. PMID- 9226195 TI - The effectiveness of deferiprone in thalassemia. PMID- 9226196 TI - Serum Ig abnormalities in mantle cell lymphoma. PMID- 9226197 TI - Elevated serum level of Fas ligand correlates with the asymptomatic stage of human immunodeficiency virus infection. PMID- 9226198 TI - Probing the cause of thyroid dysgenesis. PMID- 9226199 TI - Comparison of 18FDG-PET with 131iodine and 99mTc-sestamibi scintigraphy in differentiated thyroid cancer. AB - 18Fluorine-fluorodeoxyglucose (FDG) positron-emission tomography (PET) has emerged as a useful method in various fields of oncology. The aim of the present study was to evaluate the clinical significance of this technique in differentiated thyroid carcinoma and to compare the results with other imaging modalities, particularly with whole-body 131iodine scintigraphy (WBS) and hexakis (2-methoxyisobutylisonitrile) (99m)technetium (I) scintigraphy (MIBI). Whole-body PET imaging using FDG was performed in 54 patients. There were 39 patients with papillary tumors and 15 patients with follicular tumors (including 3 Hurthle-cell carcinomas). Primary tumor stage (pT) was pT1 in 5 cases, pT2 in 19 cases, pT3 in 2 cases, pT4 in 24 cases, and unknown in 4 cases, respectively. Finally, for each case an overall clinical evaluation was done including histology, cytology, thyroglobulin level, sonography, computed tomography, magnetic resonance imaging, and subsequent clinical course, to allow a comparison with functional imaging results. Compared with WBS, FDG-PET gave different results in the majority of cases with recurrence/metastases (11 FDG-true-positive/WBS-negative tumor sites and 8 WBS-true-positive/FDG-negative tumor sites). In 7 patients with recurrence/metastases, FDG-PET and WBS gave corresponding results (10 sites). In 28 patients, FDG-PET and WBS were normal (including 2 false-negative cases). MIBI was performed in 44 cases. FDG-PET was better correlated to MIBI (congruent positive results in 13 tumor sites) than to WBS. Compared with MIBI, FDG-PET was superior in 5 cases (including 3 patients with distant metastases). Two FDG negative/MIBI-positive tumors were observed. Different tracer uptake mechanisms have to be considered regarding "nonspecific" tumor imaging with FDG-PET or MIBI. Nevertheless, since spatial resolution with respect to tomographic imaging is inferior with single photon emission computer tomography (SPECT) using MIBI, the observed higher sensitivity of PET might be due to the higher spatial resolution of this method. As far as grading could be obtained, FDG-PET seemed to be more sensitive than WBS in high-grade tumors, whereas WBS was positive predominantly in low-grade carcinomas, although statistical significance could not be reached. The results prove the clinical usefulness of FDG-PET and MIBI for detection of 131iodine-negative tumor tissue in differentiated thyroid cancer. PMID- 9226200 TI - Telomerase activity in benign and malignant thyroid tumors. AB - Thyroid nodules are found in 5% to 10% of the population. While these nodules carry only a 5% to 10% risk of malignancy, tests that complement fine-needle aspiration (FNA) cytology in preoperative diagnosis and risk stratification are lacking. Telomerase is a ribonucleoprotein polymerase with activity found in many malignant tissues, but absent from most normal adult tissue. In this study, we have investigated telomerase activity in 24 thyroid tumors, 14 matched adjacent thyroid tissues, and 3 chronic thyroiditis tissue samples. Using a telomeric repeat amplification protocol (TRAP) assay on frozen tissue, telomerase activity was detected in 11 of 20 thyroid carcinomas, including 10 of 14 papillary carcinomas and a Hurthle cell carcinoma. Telomerase activity was not detected in 4 benign adenomas, 3 follicular carcinomas, or a single case each of medullary and anaplastic thyroid carcinoma. Telomerase activity was detected in 3 of 14 samples of adjacent thyroid tissue from patients with thyroid tumors. Interestingly, all 3 cases of adjacent thyroid tissue that tested positive had a moderate to marked degree of chronic inflammation. In addition, 3 of 3 samples from chronic thyroiditis specimens tested positive for telomerase activity. When tumor invasiveness (vascular and/or capsular) was compared with telomerase activity in papillary carcinomas, only 1 of 4 telomerase-negative tumors was invasive, while 6 of 10 of telomerase-positive tumors were invasive. Moreover, 6 of 7 invasive papillary carcinomas had telomerase activity. In summary, this is the first report of telomerase activity in thyroid tissue and nodules. This activity was detected in a large percentage of papillary thyroid carcinomas, but not benign adenomas, follicular carcinomas, or most normal thyroid tissue. Telomerase activity may also correlate with tumor invasiveness. Further studies will focus on larger numbers of tumors, metastatic tissue, and undifferentiated carcinomas, as well as application of this assay to products from fine-needle aspirates as a potential diagnostic and prognostic marker in thyroid neoplasms. PMID- 9226201 TI - Is serum thyroglobulin a useful marker for thyroid cancer in patients who have not had ablation of residual thyroid tissue? AB - Serum thyroglobulin has been measured sequentially in 47 patients with differentiated thyroid cancer who had been treated by surgery and I-thyroxine, but no radioiodine (131I). In 39 of the patients, the operation was subtotal or total thyroidectomy, and in 8 patients it was lobectomy, or lobectomy plus isthmusectomy. In the first group of patients, serum thyroglobulin (Tg) was consistently undetectable in 62% and less than 5 ng/mL in 85%. In the latter group only 1 patient had undetectable Tg values and 2 (25%) had values less than 5 ng/mL. Even in those with higher Tg values, the levels remained constant within a narrow range, provided thyrotropin (TSH) was not high. Serum Tg can be used in conjunction with clinical evaluation in follow-up of patients with differentiated thyroid cancer who have had an operation consisting of more than lobectomy and isthmusectomy and have not been treated with 131I. Its use in patients with lesser operations has to be interpreted in relation to the amount of thyroid left and to the consistency of the results with time. PMID- 9226202 TI - Expression patterns of extracellular matrix components in native and cultured normal human thyroid tissue and in human toxic adenoma tissue. AB - The extracellular matrix (ECM) and basement membranes (BM, a specialized form of ECM) greatly influence proliferation, differentiation, and function of cells and the structure of tissues. While a considerable amount of information is available on thyroid cellular proliferation, differentiation and function, much less is known about thyroid ECM and BM. In this study the presence of the ECM/BM components fibronectin, collagen IV, alpha1, beta1, gamma1 laminin, several laminin variants, osteonectin, and perlecan was demonstrated in cryosections of nonadenomatous and toxic adenoma human thyroid tissue. Also, positive immunohistochemical staining for collagen IV, laminin, perlecan, and fibronectin was obtained in sections of human thyroid tissue cultured in a three-dimensional (alginate) culture system. The present study provides methods and data that will facilitate the investigation of the interaction between cells and ECM in thyroid tissue. PMID- 9226203 TI - Efficacy of low doses of radioiodine in the treatment of autonomous thyroid nodules: importance of dose/area ratio. AB - Radioiodine (131I) represents an interesting alternative to surgery in the treatment of autonomously functioning thyroid nodules (AFTN), but leads to a significant incidence of hypothyroidism when high doses are used. Over 4 years, we have treated 40 patients (hyperthyroid [Plummer's disease]: 6, single hot nodules with undetectable thyrotropin [TSH] and normal serum free thyroxine [FT4]: 34), 34 single hot nodules with undetectable thyrotropin TSH and normal serum free thyroxine [FT4] with 131I. The dose level was neither related to the concentration of FT4 nor to the iodine uptake on thyroid scintigram. Retrospectively we measured the nodule's area on the scan and calculated the dose/area ratio (DAR). Three months after treatment, 30 patients were euthyroid, 9 were still hyperthyroid, and 1 was hypothyroid. The mean DAR of the euthyroid patients was twofold higher than for the hyperthyroid subjects (1.4 +/- 0.8 vs. 0.7 +/- 0.3 mCi/cm2; p = .003) and one-half the DAR for the hypothyroid patient (2.82 mCi/cm2). Twenty of the 30 euthyroid patients had received a dose higher than 1 mCi/cm2 and 7 of 9 hyperthyroid patients had received a dose lower than 1 mCi/cm2. (chi2 = 12.9; p = .02). The initial values of T4, TSH, and dose level of patients who were euthyroid or hyperthyroid at 3 months were not different. These data suggest that the efficacy of 131I for treating AFTN depends on the DAR, rather than the initial T4 value or the 131I uptake. A DAR between 1 and 1.5 mCi/cm2 seems to be optimal and avoids hypothyroidism. PMID- 9226204 TI - GLUT1 glucose transporter expression in benign and malignant thyroid nodules. AB - Malignant cells exhibit increased rates of glycolysis and glucose uptake, the latter of which is mediated by glucose transport proteins. Because several types of cancer have been shown to express high levels of the GLUT1 glucose transporter isoform, we hypothesized that expression of GLUT1 might distinguish malignant from benign thyroid tissue. Archival thyroid tissue obtained at surgery was immunostained for GLUT1 protein. There were 38 benign cases (24 follicular adenoma, 1 Hurthle cell adenoma, 8 nodular goiter, 3 Hashimoto's thyroiditis, 2 Graves' disease) and 28 cases of thyroid cancer (17 papillary and its follicular variant, 6 follicular, 1 Hurthle cell, 2 anaplastic, 2 medullary). Normal thyroid tissue adjacent to nodules showed no thyrocyte staining in any case. No GLUT1 staining was seen in thyrocytes in benign nodular tissue, except for a single case of Hashimoto's thyroiditis in which a few Hurthle cells showed weak staining. Among the thyroid cancers, 13 of 28 (46%) showed tumor cell GLUT1 staining in at least some areas. This included 9 of 17 cases of papillary carcinoma and its follicular variant, 2 of 6 cases of follicular carcinoma and 2 of 2 cases of anaplastic carcinoma. Tumor cell GLUT1 staining was seen in two patterns: circumferential plasma membrane staining focally within the tumor, or asymmetric staining of the basilar aspect of tumor cells adjacent to stroma in some cases of papillary carcinoma. We conclude that GLUT1 expression is frequently detectable by immunostaining in thyroid cancer, but not in benign nodules or normal thyroid. GLUT1 expression may be a clinically useful molecular marker for thyroid cancer. PMID- 9226205 TI - Clinical features of patients with Graves' disease undergoing remission after antithyroid drug treatment. AB - The clinical course of 306 Graves' patients treated with methimazole (MMI) was reviewed with the aim of establishing criteria able to predict remission of hyperthyroidism after medical treatment. One hundred and ninety-four (149 females, 45 males) of 306 (63.4%) patients had relapse of hyperthyroidism after antithyroid drug (ATD) withdrawal. Relapse was more frequent during the first months of the follow-up, but still it was observed 3 years after MMI withdrawal. The relapse rate was dependent on the age of the patient, the size of goiter, and the level of TSH-receptor antibody (TRAb) at diagnosis, being observed in 40 of 47 (85%) patients with high (> 30 U/L) TRAb level and in 54 of 101 (53%) patients with low TRAb level (< or = 30 U/L; p <.0002). Remission was more frequent (43.3%) in patients having the combination goiter size < or = 40 mL, TRAb level < or = 30 U/L, than in patients with goiter size > 40 mL and high TRAb levels (9%). In the subgroup of patients with the combination: goiter < or = 40 mL- TRAb < or = 30 U/L - age at onset > 40 years, the remission rate was 80%, and all relapses occurred within the first 9 months after MMI withdrawal. In conclusion, our study confirms that hyperthyroidism relapses in the majority of patients with Graves' disease treated with ATD. Among different clinical and laboratory features, age at onset of hyperthyroidism, goiter size and TRAb level are particularly helpful in identifying those patients who are more prone to undergo a remission of hyperthyroidism after medical treatment and may be useful to select the minority of Graves' patients who will benefit from antithyroid drug treatment as a first choice. PMID- 9226206 TI - Absence of mutations in the gene encoding thyroid transcription factor-1 (TTF-1) in patients with thyroid dysgenesis. AB - Thyroid transription factor-1 (TTF-1) is a homeodomain-containing nuclear transcription factor, important in regulation of the thyroid-specific genes thyroglobulin (Tg), thyroperoxidase (TPO), and thyrotropin receptor (TSHR). TTF-1 is an early biochemical marker of thyroid differentiation, essential for thyroid development and maintenance of the thyroid differentiated state. It is possible that mutations in titf1 gene encoding TTF-1 could result in failure of the thyroid gland to develop. Single strand conformation polymorphism (SSCP) was used to detect the presence of titf1 gene mutation in a group of 15 patients with congenital hypothyroidism. The etiology of the congenital hypothyroidism included thyroid agenesis (9), sublingual ectopic thyroid (4), and severe hypoplasia (2). The analysis did not identify any titf1 gene mutation, among these patients. These results rule out the presence of titf1 mutations, at least in the coding region, in our thyroid dysgenesis patients. Mutations in titf1 coding region may be an extremely rare event, and was not detected in our small sample size or, alternatively, such a mutant might even be viable since TTF-1 plays an important role in lung, brain, and pituitary development. PMID- 9226207 TI - Mutations in the gene encoding thyroid transcription factor-1 (TTF-1) are not a frequent cause of congenital hypothyroidism (CH) with thyroid dysgenesis. AB - Permanent congenital hypothyroidism (CH) has an incidence of 1/3000-4000 newborns and is among the most frequent cause of mental retardation and neurological alterations in children. In 80% to 85% of cases CH is associated with thyroid dysgenesis. A group of 61 patients with CH (22 with agenesis, 18 with ectopy, 1 with hypoplasia, and 20 cases with CH without thyroid enlargement but not further characterized) and 30 normal subjects were examined for the presence of mutations in the gene encoding the thyroid transcription factor 1 (TTF-1). The coding region of the TTF-1 gene was analyzed in all cases by the single stranded conformational polymorphism (SSCP) and no mutations were detected. Direct sequencing also carried out in patients with thyroid agenesis confirmed the absence of mutations or polymorphisms in the TTF-1 gene. The absence of mutations in the TTF-1 gene in our samples indicates that the mutations in the TTF-1 gene are not a frequent cause of CH. PMID- 9226208 TI - Behavioral effects of liothyronine (L-T3) in children with attention deficit hyperactivity disorder in the presence and absence of resistance to thyroid hormone. AB - Evidence that the thyroid may play a role in the pathogenesis of attention deficit hyperactivity disorder (ADHD) comes from observations that 48% to 73% of children with the syndrome of resistance to thyroid hormone (RTH) have ADHD. Casual observations in subjects with RTH have suggested that treatment with thyroid hormone may improve the symptoms of ADHD. The aim of this study was to determine whether thyroid hormone has a beneficial effect on the behavior of children with RTH. A prospective, randomized, double-blinded, placebo-controlled, cross-over study was conducted to evaluate the effect of the rapid acting thyroid hormone, liothyronine (L-T3), on the behavior of 8 children with ADHD + RTH, and 9 children with ADHD and normal thyroid function (ADHD Only). Parent and teacher ratings of hyperactivity (Conners scale) and a computerized continuous performance test (CPT) were used as objective measures of hyperactivity, attention and impulsivity. L-T3 had no effect on Conners Hyperactivity Index in 7 of 9 children with ADHD Only; it caused improvement and deterioration in 1 subject each. In contrast, the rating in 5 of 8 subjects with ADHD + RTH showed improvement, whereas 3 of 8 subjects remained unchanged. L-T3 was associated with increased commission errors in 5 of 8 children with ADHD Only and decreased commission errors in 4 of 7 with ADHD + RTH. In children with RTH and ADHD, particularly those that exhibit hyperactivity, L-T3 in supraphysiological doses may be beneficial in reducing hyperactivity and impulsivity. In the majority of children with ADHD who do not have RTH, L-T3 treatment has no effect or may be detrimental. PMID- 9226209 TI - Routine skin cleansing with povidone-iodine is not a common cause of transient neonatal hypothyroidism in North America: a prospective controlled study. AB - A high incidence of transient neonatal hypothyroidism has been observed in premature infants after routine skin cleansing with iodine. Because these reports have been predominantly from Europe, a borderline, iodine-deficient area, we wished to determine whether this was also true in North America, an iodine sufficient area. A prospective, controlled study was performed in premature babies < or = 36 weeks gestation admitted to a neonatal intensive care nursery. Thyroxine (T4) and thyrotropin (TSH) were measured at day 1, days 4 to 6, and 10 to 12 after skin preparation with iodine or with a noniodine-containing antiseptic solution (chlorhexidine) that served as control. If repeat cleansing was required, this sequence was repeated. Urinary iodine was quantitated on days 1 to 3 to estimate iodine exposure. There was no difference in the mean T4 concentration at any of the time points evaluated nor in the incidence of transient hypothyroidism between the iodine-exposed (2/17) and control babies (0/14) despite urinary iodine excretion up to 88 times the control value. Unexpectedly 5 iodine-exposed but 0 control babies developed severe hypothyroxinemia (T4 < 40 nmol/L), compatible with the sick euthyroid syndrome; one of them died. We conclude that, unlike in Europe, transient hypothyroidism is not a common sequela of routine skin cleansing with iodine in premature newborn infants in North America. This difference in incidence may be due to prior iodine status. Whether excessive iodine absorption in premature infants is associated with thyroid-independent toxic effects remains to be clarified. PMID- 9226210 TI - A longitudinal study of changes in body mass index and total body composition after radioiodine treatment for thyrotoxicosis. AB - Patients treated for thyrotoxicosis often complain of increases in body weight after treatment of their thyroid disorder. The objective of this study was to define the extent of changes in body mass and composition following treatment of thyrotoxicosis with radioiodine 131I. We prospectively measured body mass index (BMI) in 75 patients (18 males, 57 females), ranging in age from 15-88 years treated for thyrotoxicosis with 131I (doses ranging from 5.2 to 25.7 mCi) between 1978 and 1994. BMI pre- and post-radioiodine treatment were recorded for up to 10 years after treatment. Body composition studies were performed on a subgroup of 9 patients (1 male, 8 females), aged 24 to 74 years treated with 6.0 to 12.2 mCi 131I. Bone mineral content, lean mass, and fat content were determined prior to and following 131I treatment through whole body scanning with a Hologic QDR 1000/W dual energy x-ray densitometer. Sustained increases in BMI averaging 2.33 kg/m2 above baseline (p < .01) were observed from the initial 6 months through the first 5 years after 131I therapy. BMI was no different from pretreatment levels over the remainder of the 10-year follow-up period. Lean mass increased significantly (p = .0004) by an average of 7.2 kg. Although both fat and mineral content appeared to increase, these changes were not statistically significant. Weight gain occurs within the first year after 131I treatment of patients with thyrotoxicosis and is predominantly due to increased lean mass (20.2%, p < .0005) although bone mineral content increased only marginally (4.62%, p = .10). PMID- 9226211 TI - Thyroid dysfunction is not associated with alterations in serum leptin levels. AB - To determine if serum leptin levels are affected by thyroid dysfunction, we measured its concentration in serum samples from 25 euthyroid controls and 25 subjects each with hypothyroidism and thyrotoxicosis collected over a 3-month period. Mean leptin levels in the euthyroid (24.1 +/- 8.3 microg/L), hypothyroid (22.7 +/- 7.0 microg/L) and thyrotoxic (23.3 +/- 4.3 microg/L) groups were not significantly different. Data were available to express leptin in terms of body mass index (BMI) in 11 euthyroid, and 6 untreated hypothyroid and thyrotoxic individuals. There was a significant positive correlation between BMI and leptin level (r = 0.60, p = .0002) for this subgroup, irrespective of their thyroid status. These data suggest that leptin levels are not affected by thyroid dysfunction. PMID- 9226212 TI - Flow-mediated, endothelium-dependent vasodilation is impaired in subjects with hypothyroidism, borderline hypothyroidism, and high-normal serum thyrotropin (TSH) values. AB - Patients with hypothyroidism are considered to have an increased risk of developing atherosclerosis; because endothelial dysfunction is an early sign of atherosclerosis, we investigated whether endothelial dysfunction is present in patients with hypothyroidism. Thirty-five subjects with various TSH levels were investigated by high-resolution ultrasound imaging of the brachial artery to assess endothelial and smooth muscle responses. Flow-mediated, endothelium dependent vasodilatation was significantly higher in subjects with TSH 0.4-2 microIU/mL (11.8 +/- 2.7%), compared with subjects with TSH 2.01-4 microIU/mL (6.8 +/- 2.9%), 4.01-10 microIU/mL (5.2 +/- 6.3%) and >10 microIU/mL (4.0 +/- 4.4%); TSH levels correlated inversely to endothelium-dependent dilatation. Thus, flow-mediated vasodilatation, a marker of endothelial function, is impaired not only in patients with mild hypothyroidism but also in subjects with "high-normal" serum TSH levels (ie, 2.01-4.0 microIU/mL) that may be characterized as possibly abnormal. PMID- 9226213 TI - Serum N-terminal pro-atrial natriuretic factor 1-98 before and during thyroxine replacement therapy in severe hypothyroidism. AB - Decreased plasma concentrations of atrial natriuretic factor (ANF) and of its N terminal prohormones have been demonstrated in severely hypothyroid patients compared with control subjects, and shown to normalize with thyroxine (T4) replacement therapy. Whether this depends on thyroid hormone deficiency exclusively or is secondary to hemodynamic changes that result from it remains a matter of debate. In a recent investigation dose-related increases in both ANF and N-terminal prohormones of ANF by T4 replacement therapy in incremental doses increased at 4-week intervals were demonstrated. It was suggested that thyroid hormones may enhance synthesis rather than release of atrial peptide hormones. The aim of the present study was to confirm this assumption in hypothyroid patients with normal cardiac performance. Serum N-terminal amino acids 1-98 (ie, pro-ANF 1-98) of pro-ANF was determined in 11 severely hypothyroid patients without pericardial effusion and with normal cardiac left ventricular function. Mean pro-ANF 1-98 concentration before T4 replacement therapy remained unchanged after 10 days on T4 (p = .12). After 2 months of therapy, mean pro-ANF 1-98 was significantly increased compared with pretreatment values (p < .003). A significant correlation to the increase in free T4 (r = 0.48, p < .01) but not to the decrease in thyrotropin (TSH) (r = -0.32, p = .09) was found. The present results indicate that thyroid hormones directly increase pro-ANF 1-98 independently of cardiac hemodynamics in the hypothyroid state. PMID- 9226214 TI - Failure to find an association between hepatitis C virus and thyroid autoimmunity. AB - A high frequency of hepatitis C antibodies has been reported from France in patients with autoimmune thyroiditis. Two cases of Hashimoto's thyroiditis in association with chronic active hepatitis and hepatitis C infection have also been reported. We have examined this potential association in 46 patients with autoimmune hypothyroidism and found that 16 apparently had hepatitis C antibodies in one of the two commercially available enzyme-linked immunosorbent assays (ELISA), but all patients were negative in a confirmatory commercially available recombinant immunoblot assay (RIBA-3) indicating that none of the patients were truly positive for hepatitis C antibodies. We also tested sera from 111 patients with proven hepatitis C infection and found no increased prevalence of thyroid autoantibodies. These results suggest that hepatitis C infection is not a risk factor for the development of thyroid autoimmunity in the United Kingdom. PMID- 9226215 TI - Hodgkin's disease treated with neck radiation is associated with increased antibody-dependent cellular cytotoxicity against human extraocular muscle cells. AB - Patients with Hodgkin's disease have higher a prevalence of thyroid function abnormalities and, perhaps, orbitopathy than the general population, but the pathophysiology of this association and its relationship to Hodgkin's disease treatment remain unclear. We analyzed the frequency of thyroid function abnormalities, autoantibodies against thyroid antigens, and autoimmunity against extraocular muscle cell antigens by Western blot analyses and antibody-dependent cellular cytotoxicity (ADCC) assays in patients with Hodgkin's disease (n = 20) and controls (n = 10). Hodgkin's disease patients were subdivided into those treated with thyroidal external beam radiation therapy (XRT, n = 15) or chemotherapy (MOPP/ABVD, n = 5). The ADCC assay against extraocular muscle cells was increased in patients with Hodgkin's disease (5.5% vs. <1.0%, p = .026) when compared with controls. In addition, Hodgkin's disease patients treated with XRT (with or without chemotherapy) had significantly higher ADCC tests than controls (9.7% vs. <1.0%, p = .010), In contrast, ADCC assays were not different between Hodgkin's disease patients treated with chemotherapy alone and controls (<1.0% vs. <1.0%, p = .53). Hodgkin's patients treated with XRT had higher ADCC assays than those treated with chemotherapy alone (p = .087), although this difference did not achieve statistical significance. Serum measurements of antithyroid peroxidase (TPO) antibodies, antithyroglobulin (Tg) antibodies, thyroid binding inhibitory immunoglobulins (TBII), and thyroid stimulating immunoglobulin (TSI) were similar in all groups. Antibodies against the 64 kDa orbital antigen were detected in 1 patient and 1 control subject. Excluding patients already treated with L-thyroxine for hypothyroidism (n = 5), free T3, but not free T4, was lower in the Hodgkin's disease group than in controls (2.2 pg/mL vs. 2.7 pg/mL, p = .008). Thyrotropin (TSH) concentrations were not statistically different between these groups. In summary, these data show: (1) ADCC against human orbital muscle cells is increased in patients with Hodgkin's disease compared with controls: (2) these differences were noted among Hodgkin's disease patients treated with thyroidal XRT, with or without chemotherapy, and not among those patients treated with chemotherapy alone; and (3) no statistically significant differences in the frequency of thyroid autoantibodies were found. These data suggest that patients with Hodgkin's disease display altered antibody-dependent immune function toward extraocular muscle cells that may possibly be related to by XRT. Larger, prospective studies assessing thyroid and orbital-related immunologic abnormalities in Hodgkin's disease are warranted. PMID- 9226216 TI - Hyperthyroidism in McCune-Albright syndrome with a review of thyroid abnormalities sixty years after the first report. AB - We present a patient with hyperthyroidism associated with McCune-Albright syndrome (MAS). MAS is a sporadic genetic disease characterized by polyostotic fibrous dysplasia, cafe au lait cutaneous spots and endocrinopathies (peripheral precocious puberty, thyroidopathies, acromegaly, etc.). It is caused by an activating mutation of the gene for the Gs alpha membrane-associated protein, which mediates the thyrotropin (TSH)-induced and other hormone-induced activation of adenylyl cyclase. A 13-month-old girl was diagnosed with MAS. Precocious puberty was treated initially with testolactone and later with oophorectomy. Subclinical hyperthyroidism was detected biochemically at birth, and 10 months later, it became clinically evident, albeit mild, with absence of goiter. A concomitant liver dysfunction precluded treatment with thionamides and she was sporadically treated with beta-blockers. The combination of increased free thyroxine (T4) and triiodothyronine (T3) with low plasma thyrotropin (TSH) levels in the absence of thyroid-stimulating autoantibodies persisted until the age of 6 years, when she was referred to our unit. Hyperthyroidism was then clinically evident with cardiac hyperactivity, and it was cured with administration of radioiodine (131I). Thyroid disease is the second most common endocrinopathy associated with MAS, and since 1936, 63 cases of thyroidopathies have been described, including 19 nodular (14 with and 5 without hyperthyroidism) and 23 diffuse (20 with and 3 without hyperthyroidism) goiters, and 18 cases of hyperthyroidism without goiter. The previously described somatic activating mutation of the gs alpha gene in the ovaries, the liver and the peripheral blood of our patient, in the absence of stigmata, autoimmunity might be incriminated for the secretory and mitotic activation of the thyroid gland. We suggest the treatment of choice of hyperthyroidism in MAS patients should be 131I administration because: (a) hyperthyroidism is very likely to recur after withdrawal of antithyroid medication; (b) the morbidity of these patients is elevated; (c) oophorectomized patients do not need to be advised to avoid procreation during the months after 131I administration; and (d) finally, even in the usual cases of hyperthyroidism in childhood, 131I treatment is becoming more popular worldwide. PMID- 9226217 TI - A case of thyrotropin-secreting pituitary microadenoma with normal thyrotropin alpha-subunit level. AB - We present a 32-year-old male with a thyrotropin (TSH)-secreting pituitary microadenoma with normal alpha-subunit (SU) and/or alpha-SU/TSH molar ratio. An interesting feature of this patient is that the size of the pituitary tumor remained unchanged during a 6-year follow-up without treatment. The tumor was clearly visualized with somatostatin receptor imaging, indicating that it was somatostatin receptor-positive. Subcutaneous injection of 100 microg octreotide acetate three times daily resulted in significant reduction of TSH and free thyroid hormones 6 weeks after initiation of treatment. However, tumor size was not changed 3 months after initiation of octreotide therapy and thyroid hormones, but not TSH level, eventually increased in spite of increasing the octreotide dosage up to 600 microg/day. This led to discontinuation of treatment. The patient responded only temporarily to octreotide in spite of somatostatin receptors. This case further demonstrates that a normal alpha-SU and/or alpha SU/TSH molar ratio cannot exclude the possibility of a TSH-secreting pituitary adenoma. PMID- 9226218 TI - Schwannoma of the neck simulating a thyroid nodule. AB - A 31-year-old man is reported with the rare occurrence of a neurogenic tumor simulating a thyroid nodule. A growing nodule of the right neck was the only clinical symptom. Ultrasonography revealed a hypoechogenic nodule on the right side of the thyroid gland. Technetium (Tc)-99m pertechnetate showed a normal thyroid scintiscan. Due to ultrasonography-guided fine-needle aspiration biopsy, a schwannoma (neurilemmoma) was suspected. Surgical intervention removed the nodule and histology confirmed a schwannoma with Antoni A structures. To avoid unnecessary or inappropriate surgical intervention in cases of hypoechogenic nodules in connection with the thyroid gland, other nonthyroidal structures that may cause hypoechogenic patterns with ultrasonography should also be considered. The key preoperative investigations in such cases are ultrasonography and sonographically guided fine-needle aspiration biopsy. PMID- 9226219 TI - Reconstitution of triiodothyronine inhibition in non-triiodothyronine-responsive thyrotropic tumor cells using transfected thyroid hormone receptor isoforms. AB - The triiodothyronine (T3) inhibitory effect on the thyrotropin (TSH)beta- and alpha-subunit genes is believed to be mediated by binding of T3 to specific nuclear receptors that are present in various isoforms. alphaTSH cells, which are derived from a pure alpha-subunit secreting thyrotropic tumor, contain the same nuclear factors that are important for alpha-subunit gene expression in TSH expressing T3-responsive thyrotropic cells (TtT97). However, as in the parent tumor, alpha-subunit expression in alphaTSH cells was not inhibited by T3, despite the presence of high-affinity nuclear T3 receptors (TRs) with a similar number of sites per cell as in TtT97. When transcripts coding for the different TR isoforms from the MGH101A tumor were analyzed by Northern blot, TR alpha1 was present, as well as the non-T3-binding variant alpha2, but transcripts encoding the opposite strand Rev-ErbAa were not detectable and neither TR beta1 nor TR beta2 mRNAs were detectable, whereas all these transcripts were detectable in TtT97 tumors. Similar findings were observed in alphaTSH cells, where TR beta1 transcripts were barely detectable in Northern blots and TR beta2 transcripts were detectable only by RT-PCR. The TR beta gene locus is present and unrearranged in the tumor genome. In transient transfection studies conducted in alphaTSH cells overexpression of either TR beta1, TR beta2, or TR alpha1 reconstituted T3-inhibition of the alpha-subunit promoter down to 40% to 50% of control. We conclude that the relative lack of TR beta gene expression correlates with unresponsiveness to T3. The alphaTSH cell line represents a unique model in which to dissect the mechanism of T3 inhibition. PMID- 9226220 TI - Hormone-activated phosphorylation of human beta1 thyroid hormone nuclear receptor. AB - To understand the role of phosphorylation in the hormone-dependent transcriptional activation of thyroid hormone receptors (TRs), the present study evaluated the effect of the thyroid hormone, 3,3',5-triiodo-L-thyronine (T3) on the phosphorylation of TR, human subtype beta1 (h-TRbeta1). The extent of phosphorylation was compared in cells cultured in T3-depleted (Td) or T3 supplemented medium (Td + T3). T3 was found to activate phosphorylation of h TRbeta1 approximately threefold. Taking into account the T3-induced fourfold downregulation in the expression of h-TRbeta1 in the same period, the specific T3 activated phosphorylation was increased approximately twelvefold. Phosphoamino acid analysis indicates that the phosphorylation of serine and threonine in a ratio of approximately 10:1 was increased approximately threefold by T3. Comparison of the [32P]-labeled tryptic maps of h-TRbeta1 phosphorylated in cells cultured in Td medium or Td + T3 medium indicates that the latter had fewer fragments and changes of intensities in several common fragments, indicating that the phosphorylation sites activated by T3-treatment differed from those of basal phosphorylation. Partial V8 and chymotrypic proteolysis indicates that h-TRbeta1 phosphorylated in cells cultured in Td + T3 medium was more resistant to proteolysis. These results indicate that T3-activated phosphorylation altered the protease susceptibility of h-TRbeta1 that could reflect structural changes in h TRbeta1. These results raise the possibility that T3-activated phosphorylation may play an important role in transcriptional activation of h-TRbeta1. PMID- 9226221 TI - Molecular analysis of the antibody response to thyroglobulin and thyroid peroxidase. AB - In this review, we discuss the latest results concerning the molecular analysis of antibodies (Ab) directed toward thyroid autoantigens. In particular, we attempt to define patterns within the Ab repertoire that correlate best to their activities. Whilst a considerable amount is now known concerning the Ab response to thyroid peroxidase (TPO), there is still much we do not understand. We review evidence for the site of interaction of TPO-reactive Ab with native TPO. The Ab responses to thyroglobulin (Tg) and, in particular, the thyroid-stimulating hormone receptor (TSH-R), are much less well characterised. In this review, we focus on the molecular analysis of the Ab response to Tg and TPO, assessing the repertoire as it is currently known. In addition, we have tried to link this information with the analysis of the epitopes recognised by the various Ab. Finally, we discuss one of the more unusual features of the thyroid Ab repertoire, the use of D-D fusion at heavy chain junctions, and questions raised by our current state of knowledge, such as the role of Ab using germline V regions in antigen recognition. PMID- 9226222 TI - Somatostatin therapy and Graves' ophthalmopathy. PMID- 9226223 TI - Renal failure from mitochondrial cytopathies. AB - Mitochondrial cytopathies are metabolic diseases, expressing mutations in nuclear DNA, punctiform mutations or depletions in mitochondrial DNA. These genetic lesions alter mitochondrial oxidative phosphorylation, with a reduction in energy produced for cell activity. Renal disease may be the first sign of mitochondrial cytopathy, or it may appear together with neurological and neuromuscular signs. Fanconi's syndrome, a benign sign of renal tubulopathy, is particularly frequent in newborns with mitochondrial cytopathy, whereas tubulointerstitial nephropathy, which affects infants and adults, is more serious because it develops into terminal uremia. Findings of hyperlactatemia and reduced enzymatic activity on the respiratory chain in tissue biopsies are of diagnostic significance in mitochondrial cytopathy. A breakthrough is being made in our understanding of genetic alterations in mitochondrial DNA, and with future therapy, the kidney, a target organ, may be safeguarded. PMID- 9226224 TI - Hypertension in pregnancy. AB - Normal pregnancy is characterized by a marked reduction in peripheral vascular resistance. Blood pressure is diminished, while cardiac output, blood volume, renal blood flow and the glomerular filtration rate are increased. These hemodynamic changes are reversed in preeclampsia-eclampsia (PE-E), a condition considered to be related to a dysfunction of the endothelium. Decreases in the production of nitric oxide (NO) and prostacyclin (in association with an increased synthesis of thromboxane A2) may play a role in the pathogenesis of PE E. Recent reports have supported the concept that an imbalance between vasodilators and vasoconstrictors may explain the clinical, laboratory and hemodynamic disturbances observed in PE-E. Particular attention has been given to the fact that a state of L-arginine NO deficiency may be present. The control of hypertension, rest, and close clinical and laboratory surveillance remain the gold standard to minimize the severity of the complications of PE-E. However, on the basis of its physiology and pathophysiology, low-dose aspirin has been recommended in pregnancies at risk to prevent or, at least, to delay the occurrence of PE-E. Although initial reports showed promising results, recent conclusions from large trials have moderated this optimism. The use of supplemental L-arginine may be considered another possibility of treatment, and experimental data have given convincing results, but there are no reports on PE E. Therefore, the practical management of PE-E requires prudence, careful follow up and prompt decisions on the precise moment for delivery (which remains the most effective therapeutic procedure). PMID- 9226225 TI - Elevated albumin excretion in nonmodulating essential hypertensive patients. AB - Nonmodulating (NM) essential hypertensives are characterized by abnormal renal and aldosterone responses to angiotensin II. Recently, hyperinsulinemia, hypercholesterolemia, and an increased prevalence of family history of hypertension and myocardial infarction have been shown in NM hypertensives. Since an elevated urinary albumin excretion (UAE) has been indicated as a negative prognostic marker for cardiovascular diseases in essential hypertensives, we evaluated UAE in 50 male patients with mild to moderate essential hypertension (mean age 46.3 +/- 4.4 years), characterized as low renin (LR) (n = 14), modulating (M) (n = 20), and NM patients (n = 16) according to their renin profile and ability to modulate the aldosterone response to a graded infusion of angiotensin II. A group of 14 healthy male subjects (mean age 43.3 +/- 3.9 years) served as control. Resulting data showed that NM had significantly higher UAE (30.7 +/- 10.7 microg/min) than controls (11.9 +/- 2.7 microg/min, p < 0.0001), LR (22.1 +/- 8.4 microg/min, p < 0.05), and M patients (19.7 +/- 6.6 microg/min, p = 0.0001) when all fed a 200-mmol NaCl/day diet. On the contrary, differences in UAE disappeared when all subjects were on a low sodium regimen (10 mmol NaCl/day). Compared to LR and M patients, the NM ones also manifested higher low density lipoprotein cholesterol levels (p < 0.05). Furthermore, these latter and UAE were positively correlated in NM patients (r = 0.579, p < 0.05) but not in the other subgroups. In conclusion, the current study demonstrates elevated UAE in NM patients, suggesting the NM phenotype is combined to an increased cardiovascular risk. PMID- 9226226 TI - Over 11 years of stable renal function after remission of nephrotic-range proteinuria in type I diabetics treated with an ACE inhibitor. AB - It has previously been considered inevitable that a progressive deterioration in renal function would occur in type I diabetics who have proteinuria in the nephrotic range. We have reviewed all type I diabetic patients presenting with nephrotic-range albuminuria to this department over a 13-year period. Of 16 patients identified, 4 have demonstrated a prolonged stability of renal function, with 2 losing their albuminuria. The latter 2 patients, who have been treated with an angiotensin-converting enzyme inhibitor for over 11 years, are presented in detail. The possible factors contributing to progression and the role of angiotensin-converting enzyme inhibitors in the treatment of advanced diabetic nephropathy are discussed. PMID- 9226227 TI - Differential cytokine regulation of complement proteins in human glomerular epithelial cells. AB - We have previously shown in inbred strains of mice which naturally develop systemic lupus erythematosus that kidney C3, C2, C4 and factor B gene expression increases coincidently with the occurrence of glomerulonephritis, suggesting that local tissue complement gene expression could contribute to the pathogenesis of immune complex injury. In this study, we investigated the synthesis of complement proteins in glomerular epithelial cells (GECs) and its regulation. Using biosynthetic labelling, immunoprecipitation and sodium dodecyl sulfate polyacrylamide gel electrophoresis, we demonstrated that GECs synthesized C1r, C1s, C1 inhibitor, C3, C2 and factor B. Interferon-gamma induced increases in the synthesis of all these proteins. Both factor B and C3 proteins were increased following addition of either IL-1beta, IL-6 or TNF-alpha to GEC cultures; however, these cytokines did not increase either C2, C1r, C1s or C1-inhibitor biosynthesis. Lipopolysaccharide affected the biosynthesis of these proteins in a similar way. A semiquantitative analysis of the mRNA expression of some of these proteins by reverse-transcriptase polymerase chain reaction showed that these cytokine effects were pretranslational as there was enhancement of factor B mRNA expression by IL-1, TNF-alpha, IFN-gamma, IL-6 and endotoxin, but only IFN-gamma enhanced C1-inhibitor and C4 mRNA expression. These results may be of significance in the immunopathogenesis of glomerulonephritis, where it is likely that local complement production in GECs is independently regulated by cytokines, derived from resident glomerular mesangial cells or infiltrating monocyte/macrophages and T cells. PMID- 9226228 TI - HIV-1 gp160 envelope protein modulates proliferation and apoptosis in mesangial cells. AB - Mesangial cell (MC) hyperplasia and accumulation of extracellular matrix are the predominant features of HIV-associated nephropathy (HIVAN). Since mice transgenic for HIV-1 genes show renal lesions mimicking HIVAN, we studied the effect of HIV 1 gp160 protein on cultured murine MC (MMC) proliferation and apoptosis. HIV-1 gp160 protein stimulated (p < 0.001) MMC proliferation when compared with control MMCs. This effect of gp160 protein peaked at a concentration of 0.01 microg/ml. MMCs treated with a higher concentration of gp160 protein (0.1 microg/ml) or for a prolonged period of time (72 h) showed apoptosis rather than cell proliferation. These studies were further confirmed by DNA fragmentation and end labeling assays. gp160 also enhanced apoptosis in human MCs. Tumor necrosis factor (TNF)-alpha enhanced (p < 0.001) MMC apoptosis, and anti-TNF-alpha antibodies inhibited gp160-induced MMC apoptosis. In addition, gp160 protein attenuated MMC expression of Bcl-2 mRNA expression. These results suggest that gp160-induced apoptosis may be affected in part by the release of TNF-alpha and associated with attenuated mRNA expression of Bcl-2 by MMCs. PMID- 9226229 TI - Creatine kinase subform analysis in hemodialysis patients without acute coronary syndromes. AB - The finding of elevated levels of creatine kinase (CK) and its myocardial isoenzyme (CK MB) in the blood of patients with acute chest pain syndromes is a key factor in making the diagnosis of acute myocardial infarction. With the widespread use of thrombolytic therapy and emergency angioplasty, the ability to make a rapid and accurate diagnosis of myocardial infarction is critical. A new rapid method for analysis of the subforms of the MB isoenzyme has been shown to be sensitive and specific for the diagnosis of myocardial infarction in the general population. Because of its rapidity it has been replacing standard analyses of total CK and CK MB in some centers in guiding the initial therapy of patients with chest pain. Levels of CK MB can be abnormally elevated in hemodialysis patients even in the absence of acute myocardial necrosis. This study was undertaken in order to determine if subform analysis of the MB isoenzyme could similarly be abnormal in hemodialysis patients without acute coronary syndromes. CK MB subforms were analyzed in 52 patients without any recent cardiac symptoms who came into the dialysis unit for a routine hemodialysis treatment. We then applied the same criteria for the diagnosis of an acute myocardial infarction as would be used clinically. In this population, the subform analysis was surprisingly consistent with myocardial infarction in approximately 29% of the patients. Thus, subform analysis appears likely to result in an erroneous diagnosis of acute myocardial infarction in patients on hemodialysis. PMID- 9226231 TI - Age-related changes of urinary nitrite/nitrate excretion in normal children. AB - We measured the urinary excretion of nitrite/nitrate, stable metabolites of nitric oxide, using the brucine method in 90 healthy normal children (47 boys and 43 girls), aged from 1.0 to 17.1 years, to establish the age-related normal range in children. The urinary nitrite/nitrate excretion was highest in the youngest children and decreased in an age-dependent manner to reach constant levels at about 12 years of age in both sexes. The data may be useful in identifying sick children with abnormal nitric oxide production. PMID- 9226230 TI - Cytoprotective effect of ulinastatin on LLC-PK1 cells treated with antimycin A, gentamicin, and cisplatin. AB - The cytoprotective effect of ulinastatin was studied in LLC-PK1 cells treated with antimycin A, gentamicin, or cisplatin. All of the three agents induced a concentration-dependent increase in the release of lactate dehydrogenase and a decrease in the amount of remaining protein. In the cell injury models treated with 1.5 microM antimycin A, 10 mM gentamicin, and 0.3 mM cisplatin, ulinastatin tended to show a cytoprotective effect at a concentration of 3,000 U/ml and provided a significant protective effect at 10,000 U/ml. LLC-PK1 cells treated with 0.3 mM cisplatin, bovine serum albumin, and alpha1-acid glycoprotein at a concentration of 3.54 mg/ml, which is a comparable protein concentration to that of 10,000 U/ml ulinastatin, showed no protective effect but rather enhanced cell injury. These results suggest that ulinastatin exerts a direct protective effect on LLC-PK1 cells against various renal toxicities. PMID- 9226232 TI - Cultures of aspiration biopsy specimens in the immunological monitoring of renal transplants. AB - OBJECTIVE: Graft-infiltrating cells (GIC) have been studied in heart, lung, and liver transplants and have been shown to have greater proliferative ability when taken from rejecting allografts. Our aim was to study GIC harvested by fine needle aspiration biopsy (FNAB) in renal transplant recipients. PATIENTS AND METHODS: 93 adult patients entered the study. The FNABs were done on the 7th, 14th, and 30th day after transplantation in stable cases and whenever a rejection crisis supervened. RESULTS: The proliferation responses of GIC were significantly higher in rejection than in stable cases during the 1st month after transplantation. The sensitivity for rejection was 96.4%, the specificity 91.3%, the negative predictive value 98.7%, and the positive predictive value was 93.3% among dysfunctioning grafts. CONCLUSIONS: The study of the proliferative capacity of graft-infiltrating cells in renal transplants is a safe and very useful immunologic monitoring tool, and it could improve the FNAB diagnostic accuracy. PMID- 9226233 TI - Therapeutic effect of a newly developed antioxidative agent (OPC-15161) on experimental immune complex nephritis. AB - The effect of a newly developed free radical scavenger (OPC-15161) on the progression of nephrotoxic serum (NTS) nephritis was evaluated. NTS nephritis rats were sacrificed immediately before and 1, 2, 3, 6, and 24 h and 13 and 19 days after intravenous injection of NTS. The tissue content of phosphatidylcholine hydroperoxide, the activity of superoxide, the activity of superoxide dismutase in the renal cortex, and the serum malondialdehyde levels were measured. The phosphatidylcholine hydroperoxide content in the renal cortex of OPC-15161-treated NTS nephritis rats was lower than that in the control rats 24 h after NTS injection. The activity of superoxide dismutase in OPC-15161 treated rats was sustained in contrast to the decrease in this activity in the control rats 6 h after injection of NTS. The effects of OPC-15161, dipyridamole, and prednisolone on NTS nephritis rats were investigated. OPC-15161 (20 mg/kg p.o.) showed a potent inhibitory effect on the urinary protein excretion, whereas dipyridamole (30 and 100 mg/kg p.o.) and prednisolone (2 mg/kg p.o.) had less suppressive effects. In view of these results, we conclude that OPC-15161 notably ameliorated the urinary protein excretion by way of the suppression of lipid peroxidation in the renal tissue of NTS nephritis rats. PMID- 9226234 TI - Effects of antihypertensive drugs or glycemic control on antioxidant enzyme activities in spontaneously hypertensive rats with diabetes. AB - The activities of glomerular intrinsic antioxidant enzymes (AOEs) were measured in a diabetic spontaneously hypertensive rat (SHR) model. The effects of antihypertensive drugs, i.e. captopril or triple therapy (hydralazine, reserpine, and hydrochlorothiazide), on glomerular intrinsic AOE activities in this model were evaluated. The effects of blood glucose control on the AOE activities were also determined. The aim of the present study was to determine whether activities of glomerular intrinsic AOEs might correlate with disease activity in diabetic SHR. This study showed a decrease of glomerular intrinsic AOE, i.e. Cu/Zn-SOD and Mn-SOD (SOD = superoxide dismutase), glutathione peroxidase, and catalase, activities in diabetic SHR. Glomerular Cu/Zn-SOD or Mn-SOD, glutathione peroxidase, and catalase activities in nondiabetic SHR were slightly lower than those in nondiabetic WKY rats. These activities in diabetic SHR were significantly improved after captopril or triple therapy or blood glucose control. The levels of urinary albumin excretion, creatinine clearance, and glomerular tuft areas in diabetic SHR were also improved after the therapy. It appears that hypertension and hyperglycemia may influence the glomerular intrinsic AOE activities, albuminuria, creatinine clearance, and glomerular tuft areas in diabetic SHR. Thus, it is indicated that control of blood pressure or blood glucose is a very important factor for preventing renal injuries in the diabetic SHR model. PMID- 9226235 TI - In vitro macro- and microautoradiographic localization of V1 and V2 receptors in the rat kidney using OPC-21268 and OPC-31260. AB - To elucidate the precise localization of vasopressin (VP) V1 and V2 receptors in the kidney, we utilized in vitro macroautoradiography (macro-ARG) and microautoradiography (micro-ARG) of these receptors in Wistar rat kidneys. This was done by using OPC-21268 and OPC-31260, two newly developed selective V1 (OPC 21268) and V2 (OPC-31260) receptor antagonists. For macro-ARG, 10-microm kidney sections were incubated with Tris-HCl buffer containing [3H]-VP with or without unlabeled ligand (VP, OPC-21268, or OPC-31260) at 20 degrees C for 40 min. These sections were then loaded into X-ray cassettes with Hyperfilm-[3H] and exposed in the dark for 2 months. The autoradiograms were quantitatively analyzed by using the research analysis system RAS 1,000; the V1 and V2 receptors were quantitated by subtracting the nonspecific binding (incubated with OPC-21268 and OPC-31260, respectively) from the total binding. To assess a more precise localization of the V1 and V2 receptors, we also investigated the micro-ARG of the renal V1 and V2 receptors by dipping the kidney section slides used for macro-ARG into a photographic emulsion and observing the receptors under light microscopy. [3H]-VP binding to the rat kidney was completely displaced by unlabeled excess VP, but not by unlabeled angiotensin II, indicating that [3H]-VP binding was specific for VP receptors. Computerized quantification showed that V2 receptors, visualized by OPC-31260, were the predominant type of VP receptor in the kidney. Conversely, V1 receptors, visualized by OPC-21268, were fewer in number. V1 receptors were partly localized to the glomerulus, cortical vessels, interstitial cells, and the medullary vessels. The V2 receptors localized to the collecting ducts and medullary tubules. Our findings indicated that renal V1 and V2 receptors can be detected by in vitro macro- and micro-ARG by using OPC-21268 and OPC-31260. PMID- 9226236 TI - Cyclosporin-A- and angiotensin-II-induced vasoreactivity in isolated glomeruli and cultured mesangial cells, 4 and 24 h after renal mass reduction: role of vasodilatory prostaglandins. AB - Subtotal nephrectomy in the rat is followed by early hypertrophy and functional adaptation of the remnant glomeruli in which mesangial cells seem to play a leading part. Using a morphoquantitative approach, we assessed the vasoconstrictive response to 1 microM cyclosporin A (CsA) or 0.1 microM angiotensin II (AII) of isolated glomeruli and cultured mesangial cells obtained from normal rats and from rats after five-sixths nephrectomy. In normal rats, a significant reduction of glomerular size was observed after the administration of vasoactive agents (-14% after 30 min exposure to CsA or AII). After five-sixths nephrectomy the vasoconstrictive response, which was preserved at the 4th hour ( 12% at the 30th minute), disappeared at the 24th hour (-1.5% at the 30th minute), but was maintained in animals pretreated with indomethacin (-10% at the 30th minute). Similar responses were observed with cultured mesangial cells at the first passage. Inhibition of prostaglandin synthesis abolishes the adaptative adjustments observed in partially nephrectomized rats during the first days after surgery. These findings suggest that renal vasodilatory prostaglandins may participate in the compensatory hemodynamic adjustments following subtotal nephrectomy. PMID- 9226237 TI - Hyperlipidemic nephropathy induced by adriamycin: effect of melatonin administration. AB - The effect of melatonin (MEL) on the nephropathy and the oxidative stress induced by a single and high dose of Adriamycin (AD) has been studied in Wistar male rats. MEL (50 microg/kg/day) was injected intraperitoneally 3 and 7 days, respectively, before and after AD injection (20 mg/kg i.p.). Trunk blood was drawn and triglycerides, total cholesterol, phospholipids, high-density lipoprotein cholesterol, urea, creatinine, total protein, lipoperoxides, and reduced glutathione (GSH) levels and catalase activity (CAT) were determined in serum. In kidney homogenates, lipoperoxides, GSH, and CAT were measured as well as total protein in urine. AD administration resulted in hyperlipidemia and high grade proteinuria and a marked increase in serum lipoperoxides, urea, and creatinine. In the kidney, the increase in lipoperoxides was accompanied by a significant decrease of GSH and CAT. The efficiency of MEL was specially remarkable in restoring GSH, CAT, and proteinuria to the levels of controls. These results confirm the involvement of free radicals in the pathogenesis of nephrotoxicity induced by AD. Likewise, they show the high antioxidative power of MEL and its marked effect on the prevention and suppression of this nephropathy. PMID- 9226238 TI - Page kidney in a hemodialyzed patient. PMID- 9226240 TI - Cost-benefit = the cost of life - the economic benefit of death. PMID- 9226239 TI - Castleman's disease in a renal allograft recipient. PMID- 9226242 TI - Is HCV-infection another reason to opt for peritoneal dialysis in end-stage renal failure? PMID- 9226241 TI - Hepatitis C virus seropositivity in glomerulonephritis patients. PMID- 9226243 TI - Urinary tract carcinoma in leprosy patients treated with dapsone for a long period. PMID- 9226244 TI - End-stage renal disease in a patient with Werner's syndrome. PMID- 9226246 TI - Metabolic alkalosis and chronic renal failure in a bulimic patient. PMID- 9226245 TI - Acute leukemia in a uremic patient undergoing erythropoietin treatment. PMID- 9226247 TI - Lipoperoxidation and glutathione-dependent enzymes in uremic anemia of CAPD patients. PMID- 9226248 TI - The activation of serum hepatocyte growth factor in acute renal failure. PMID- 9226249 TI - Serum osteocalcin in children with chronic renal failure. PMID- 9226250 TI - Hyperbilirubinemia in a renal transplant patient due to cyclosporin A therapy. PMID- 9226251 TI - Alterations in induced potassium calcium efflux in the erythrocytes of patients with autosomal dominant polycystic kidney disease and hypertension. PMID- 9226252 TI - Vascular access infection associated with methicillin-resistant Staphylococcus aureus nasal carriage in a hemodialysis patient. PMID- 9226253 TI - Depletion of pre-16S rRNA in starved Escherichia coli cells. AB - Specific hybridization assays for intermediates in rRNA synthesis (pre-rRNA) may become useful for monitoring the growth activity of individual microbial species in complex natural systems. This possibility depends upon the assumption that rRNA processing in microbial cells continues after growth and pre-rRNA synthesis cease, resulting in drainage of the pre-rRNA pool. This is not the case in many eukaryotic cells, but less is known about the situation in bacteria. Therefore, we used DNA probes to measure steady-state cellular pre-16S rRNA pools during growth state transitions in Escherichia coli. Pre-16S rRNA became undetectable when cells entered the stationary phase on rich medium and was replenished upon restoration of favorable growth conditions. These fluctuations were of much greater magnitude than concurrent fluctuations in the mature 16S rRNA pool. The extent of pre-16S rRNA depletion depended upon the circumstances limiting growth. It was significantly more pronounced in carbon-energy-starved cells than in nitrogen-starved cells or in cells treated with energy uncouplers. In the presence of the transcriptional inhibitor rifampin, rates of pre-16S rRNA depletion in carbon-energy-starved cells and nitrogen-starved cells were similar, suggesting that the difference between these conditions resides primarily at the level of pre-rRNA synthesis. Chloramphenicol, which inhibits the final steps in rRNA maturation, halted pre-16S rRNA depletion under all conditions. The data show that E. coli cells continue to process pre-rRNA after growth and rrn operon transcription cease, leading to drainage of the pre-rRNA pool. This supports the feasibility of using pre-rRNA-targeted probes to monitor bacterial growth in natural systems, with the caveat that patterns of pre-rRNA depletion vary with the conditions limiting growth. PMID- 9226254 TI - Characterization of the alginate biosynthetic gene cluster in Pseudomonas syringae pv. syringae. AB - Alginate, a copolymer of D-mannuronic acid and L-guluronic acid, is produced by a variety of pseudomonads, including Pseudomonas syringae. Alginate biosynthesis has been most extensively studied in P. aeruginosa, and a number of structural and regulatory genes from this species have been cloned and characterized. In the present study, an alginate-defective (Alg-) mutant of P. syringae pv. syringae FF5 was shown to contain a Tn5 insertion in algL, a gene encoding alginate lyase. A cosmid clone designated pSK2 restored alginate production to the algL mutant and was shown to contain homologs of algD, alg8, alg44, algG, algX (alg60), algL, algF, and algA. The order and arrangement of the structural gene cluster were virtually identical to those previously described for P. aeruginosa. Complementation analyses, however, indicated that the structural gene clusters in P. aeruginosa and P. syringae were not functionally interchangeable when expressed from their native promoters. A region upstream of the algD gene in P. syringae pv. syringae was shown to activate the transcription of a promoterless glucuronidase (uidA) gene and indicated that transcription initiated upstream of algD as described for P. aeruginosa. Transcription of the algD promoter from P. syringae FF5 was significantly higher at 32 degrees C than at 18 or 26 degrees C and was stimulated when copper sulfate or sodium chloride was added to the medium. Alginate gene expression was also stimulated by the addition of the nonionic solute sorbitol, indicating that osmolarity is a signal for algD expression in P. syringae FF5. PMID- 9226255 TI - Characterization of Lactococcus lactis UV-sensitive mutants obtained by ISS1 transposition. AB - Studies of cellular responses to DNA-damaging agents, mostly in Escherichia coli, have revealed numerous genes and pathways involved in DNA repair. However, other species, particularly those which exist under different environmental conditions than does E. coli, may have rather different responses. Here, we identify and characterize genes involved in DNA repair in a gram-positive plant and dairy bacterium, Lactococcus lactis. Lactococcal strain MG1363 was mutagenized with transposition vector pG+host9::ISS1, and 18 mutants sensitive to mitomycin and UV were isolated at 37 degrees C. DNA sequence analyses allowed the identification of 11 loci and showed that insertions are within genes implicated in DNA metabolism (polA, hexB, and deoB), cell envelope formation (gerC and dltD), various metabolic pathways (arcD, bglA, gidA, hgrP, metB, and proA), and, for seven mutants, nonidentified open reading frames. Seven mutants were chosen for further characterization. They were shown to be UV sensitive at 30 degrees C (the optimal growth temperature of L. lactis); three (gidA, polA, and uvs-75) were affected in their capacity to mediate homologous recombination. Our results indicate that UV resistance of the lactococcal strain can be attributed in part to DNA repair but also suggest that other factors, such as cell envelope composition, may be important in mediating resistance to mutagenic stress. PMID- 9226256 TI - Regulation of the formate dehydrogenase gene, FDH1, in the methylotrophic yeast Candida boidinii and growth characteristics of an FDH1-disrupted strain on methanol, methylamine, and choline. AB - The structural gene (FDH1) coding for NAD(+)-dependent formate dehydrogenase (FDH) was cloned from a genomic library of Candida boidinii, and the FDH1 gene was disrupted in the C. boidinii genome (fdh1 delta) by one-step gene disruption. In a batch culture experiment, although the fdh1 delta strain was still able to grow on methanol, its growth was greatly inhibited and a toxic level of formate was detected in the medium. In a methanol-limited chemostat culture at a low dilution rate (0.03 to 0.05 h[-1]), formate was not detected in the culture medium of the fdh1 delta strain; however, the fdh1 delta strain showed only one fourth of the growth yield of the wild-type strain. Expression of FDH1 was found to be induced by choline or methylamine (used as a nitrogen source), as well as by methanol (used as a carbon source). Induction of FDH1 was not repressed in the presence of glucose when cells were grown on methylamine, choline, or formate, and expression of FDH1 was shown to be regulated at the mRNA level. Growth on methylamine or choline as a nitrogen source in a batch culture was compared between the wild type and the fdh1 delta mutant. Although the growth of the fdh1 delta mutant was impaired and the level of formate was higher in the fdh1 delta mutant than in the wild-type strain, the growth defect caused by FDH1 gene disruption was small and less severe than that caused by growth on methanol. As judged from these results, the main physiological role of FDH with all of the FDH1-inducing growth substrates seems to be detoxification of formate, and during growth on methanol, FDH seems to contribute significantly to the energy yield. PMID- 9226257 TI - Characterization of a second lysine decarboxylase isolated from Escherichia coli. AB - We report here on the existence of a new gene for lysine decarboxylase in Escherichia coli K-12. The hybridization experiments with a cadA probe at low stringency showed that the homologous region of cadA was located in lambda Kohara phage clone 6F5 at 4.7 min on the E. coli chromosome. We cloned the 5.0-kb HindIII fragment of this phage clone and sequenced the homologous region of cadA. This region contained a 2,139-nucleotide open reading frame encoding a 713-amino acid protein with a calculated molecular weight of 80,589. Overexpression of the protein and determination of its N-terminal amino acid sequence defined the translational start site of this gene. The deduced amino acid sequence showed 69.4% identity to that of lysine decarboxylase encoded by cadA at 93.7 min on the E. coli chromosome. In addition, the level of lysine decarboxylase activity increased in strains carrying multiple copies of the gene. Therefore, the gene encoding this lysine decarboxylase was designated Idc. Analysis of the lysine decarboxylase activity of strains containing cadA, ldc, or cadA ldc mutations indicated that ldc was weakly expressed under various conditions but is a functional gene in E. coli. PMID- 9226258 TI - Cloning and characterization of the region III flagellar operons of the four Shigella subgroups: genetic defects that cause loss of flagella of Shigella boydii and Shigella sonnei. AB - To detect genetic defects that might have caused loss of flagella in Shigella boydii and Shigella sonnei, the region III flagellar (fli) operons were cloned from certain strains and analyzed with reference to the restriction maps and genetic maps of Escherichia coli fli operons. S. boydii NCTC9733 (strain C5 in this paper) had the 988-bp internal deletion in the fliF gene that encodes a large substructural protein of the basal body. Two strains (C1 and C8) had deletions of the entire fliF operon, and the remaining three (C3, C4, and C9) differed in the size of the restriction fragments carrying the fliF and fliL operons. Loss of flagella in S. boydii appears to originate in some defect in the fliF operon. S. sonnei IID969 lacked the fliD gene and, in place of it, carried two IS600 elements as inverted repeats. Genes downstream from fliD were not detected in the cloned fragment despite its large size but did appear elsewhere in the chromosome. The fliD gene encodes a cap protein of the flagellar filament, and its deletion results in overexpression of class 3 operons by the increased amount of FliA (sigmaF) caused by the excess export of the anti-sigma factor FlgM. Three other strains also had the fliD deletion, and two of them had another deletion in the fliF-fliG-fliH region. The fliD deletion might be the primary cause of loss of flagella in S. sonnei. The lack of FliF or FliD in each subgroup is discussed in connection with the maintenance of virulence and bacterial growth. We also discuss the process of loss of flagella in relation to transposition of IS elements and alterations of the noncoding region, which were found to be common to at least three subgroups. PMID- 9226259 TI - The kdp system of Clostridium acetobutylicum: cloning, sequencing, and transcriptional regulation in response to potassium concentration. AB - The complete sequence of the kdp gene region of Clostridium acetobutylicum has been determined. This part of the chromosome comprises two small open reading frames (orfZ and orfY), putatively encoding hydrophobic peptides, and the genes kdpA, kdpB, kdpC, and kdpX, followed by an operon encoding a pair of sensor effector regulatory proteins (KdpD and KdpE). Except for orfZ, orfY, and kdpX, all genes showed significant homology to the kdp genes of Escherichia coli, encoding a high-affinity potassium transport ATPase and its regulators. The complete genome sequence of Synechocystis sp. strain PCC 6803 and a recently published part of the Mycobacterium tuberculosis genome indicate the existence of a kdp system in these organisms as well, but all three systems comprise neither a second orf upstream of kdpA nor an additional kdpX gene. Expression of the clostridial kdp genes, including the unique kdpX gene, was found to be inducible by low potassium concentrations. A transcription start point could be mapped upstream of orfZ. A promoter upstream of kdpD was active only under noninducing conditions. Lowering the potassium content of the medium led to formation of a common transcript (orfZYkdpABCXDE), with a putative internal RNase E recognition site, which could be responsible for the instability of the common transcript. Except for the two small peptides, all gene products could be detected in in vitro transcription-translation experiments. PMID- 9226260 TI - Functional complementation of an Escherichia coli gap mutant supports an amphibolic role for NAD(P)-dependent glyceraldehyde-3-phosphate dehydrogenase of Synechocystis sp. strain PCC 6803. AB - The gap-2 gene, encoding the NAD(P)-dependent D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH2) of the cyanobacterium Synechocystis sp. strain PCC 6803, was cloned by functional complementation of an Escherichia coli gap mutant with a genomic DNA library; this is the first time that this cloning strategy has been used for a GAPDH involved in photosynthetic carbon assimilation. The Synechocystis DNA region able to complement the E. coli gap mutant was narrowed down to 3 kb and fully sequenced. A single complete open reading frame of 1,011 bp encoding a protein of 337 amino acids was found and identified as the putative gap-2 gene identified in the complete genome sequence of this organism. Determination of the transcriptional start point, identification of putative promoter and terminator sites, and orientation of the truncated flanking genes suggested the gap-2 transcript should be monocystronic, a possibility further confirmed by Northern blot studies. Both natural and recombinant homotetrameric GAPDH2s were purified and found to exhibit virtually identical physicochemical and kinetic properties. The recombinant GAPDH2 showed the dual pyridine nucleotide specificity characteristic of the native cyanobacterial enzyme, and similar ratios of NAD- to NADP-dependent activities were found in cell extracts from Synechocystis as well as in those from the complemented E. coli clones. The deduced amino acid sequence of Synechocystis GAPDH2 presented a high degree of identity with sequences of the chloroplastic NADP-dependent enzymes. In agreement with this result, immunoblot analysis using monospecific antibodies raised against GAPDH2 showed the presence of the 38-kDa GAPDH subunit not only in crude extracts from the gap-2-expressing E. coli clones and all cyanobacteria that were tested but also in those from eukaryotic microalgae and plants. Western and Northern blot experiments showed that gap-2 is conspicuously expressed, although at different levels, in Synechocystis cells grown in different metabolic regimens, even under chemoheterotrophic conditions. A possible amphibolic role of the cyanobacterial GAPDH2, namely, anabolic for photosynthetic carbon assimilation and catabolic for carbohydrate degradative pathways, is discussed. PMID- 9226262 TI - Microbial degradation of chloroaromatics: use of the meta-cleavage pathway for mineralization of chlorobenzene. AB - Pseudomonas putida GJ31 is able to simultaneously grow on toluene and chlorobenzene. When cultures of this strain were inhibited with 3-fluorocatechol while growing on toluene or chlorobenzene, 3-methylcatechol or 3-chlorocatechol, respectively, accumulated in the medium. To establish the catabolic routes for these catechols, activities of enzymes of the (modified) ortho- and meta-cleavage pathways were measured in crude extracts of cells of P. putida GJ31 grown on various aromatic substrates, including chlorobenzene. The enzymes of the modified ortho-cleavage pathway were never present, while the enzymes of the meta-cleavage pathway were detected in all cultures. This indicated that chloroaromatics and methylaromatics are both converted via the meta-cleavage pathway. Meta cleavage of 3-chlorocatechol usually leads to the formation of a reactive acylchloride, which inactivates the catechol 2,3-dioxygenase and blocks further degradation of catechols. However, partially purified catechol 2,3-dioxygenase of P. putida GJ31 converted 3-chlorocatechol to 2-hydroxy-cis,cis-muconic acid. Apparently, P. putida GJ31 has a meta-cleavage enzyme which is resistant to inactivation by the acylchloride, providing this strain with the exceptional ability to degrade both toluene and chlorobenzene via the meta-cleavage pathway. PMID- 9226261 TI - Bacillus subtilis CcdA-defective mutants are blocked in a late step of cytochrome c biogenesis. AB - Cytochromes of the c type contain covalently bound heme. In bacteria, they are located on the outside of the cytoplasmic membrane. Cytochrome c synthesis involves export of heme and apocytochrome across the cytoplasmic membrane followed by ligation of heme to the polypeptide. Using radioactive protoheme IX produced in Escherichia coli, we show that Bacillus subtilis can use heme from the growth medium for cytochrome c synthesis. The B. subtilis ccdA gene encodes a 26-kDa integral membrane protein which is required for cytochrome c synthesis (T. Schiott et al., J. Bacteriol. 179:1962-1973, 1997). In this work, we analyzed the stage at which cytochrome c synthesis is blocked in a ccdA deletion mutant. The following steps were found to be normal in the mutant: (i) transcription and translation of cytochrome c structural genes, (ii) translocation of apocytochrome across the cytoplasmic membrane, and (iii) heme transport from the cytoplasm to cytochrome polypeptide on the outer side of the cytoplasmic membrane. It is concluded that CcdA is required for a late step in the cytochrome c synthesis pathway. PMID- 9226263 TI - Iron-regulated excretion of alpha-keto acids by Salmonella typhimurium. AB - Excretion of alpha-keto acids by clinical isolates and laboratory strains of Salmonella typhimurium was determined by high-performance liquid chromatography analysis of culture supernatants. The levels of excretion increased markedly with increasing iron stress imposed by the presence of alpha,alpha'-dipyridyl or conalbumin in the medium. The major product was pyruvic acid, but significant concentrations of alpha-ketoglutaric acid, alpha-ketoisovaleric acid, and alpha ketoisocaproic acid were also observed. Maximal excretion occurred at iron stress levels that initially inhibited bacterial growth; the concentration of alpha,alpha'-dipyridyl at which this was observed differed between strains depending on their ability to secrete and utilize siderophores, suggesting that the intracellular iron status was important in determining alpha-keto acid excretion. However, prolonged incubation of the siderophore-deficient S. typhimurium strain enb-7 under conditions of high iron stress resulted in significant delayed bacterial growth, promoted by tonB-dependent uptake of iron complexed with the high accumulated levels of pyruvic acid and other alpha-keto acids. Strain RB181, a fur derivative of enb-7, excreted massive amounts of alpha keto acids into the culture medium even in the absence of any iron chelators (the concentration of pyruvic acid, for example, was >25 mM). Moreover, RB181 was able to grow and excrete alpha-keto acids in the presence of alpha,alpha'-dipyridyl at concentrations threefold greater than that which inhibited the growth of enb-7. PMID- 9226264 TI - The 75-kilodalton antigen of Bartonella bacilliformis is a structural homolog of the cell division protein FtsZ. AB - A genomic library of Bartonella bacilliformis was constructed and screened with human anti-Bartonella serum from a patient with the chronic, verruga peruana phase of bartonellosis. An immunoreactive clone isolated from this library was found to code for a 591-amino-acid protein with a high degree of sequence similarity to the FtsZ family of proteins. The degree of amino acid identity between the B. bacilliformis protein (FtsZ[Bb]) and the other FtsZ proteins is especially pronounced over the N-terminal 321 amino acids (N-terminal domain) of the sequence, with values ranging from 45% identity for the homolog from Micrococcus luteus (FtsZ[Ml]) to 91% identity for the homolog from Rhizobium melliloti, (FtsZ[Rm1]). All of the functional domains required for FtsZ activity are conserved in FtsZ(Bb) and are located within the N-terminal domain of the protein. FtsZ(Bb) is approximately twice as large as most of the other FtsZ proteins previously reported, a property it shares with FtsZ(Rm1). Like the Rhizobium homolog, FtsZ(Bb) has a C-terminal region of approximately 256 amino acids that is absent in the other FtsZ proteins. Evidence is presented that implicates this region in the protein's antigenicity and suggests that, unlike most other FtsZ homologs, FtsZ(Bb) is at least partly exposed at the cell surface. PCR analysis revealed that an ftsZ gene similar in size to the B. bacilliformis gene is present in Bartonella henselae, a bacterium that is closely related to B. bacilliformis. PMID- 9226265 TI - Occurrence of deletions, associated with genetic instability in Streptomyces ambofaciens, is independent of the linearity of the chromosomal DNA. AB - The chromosomal structures of mutant strains of Streptomyces ambofaciens which have arisen from genetic instability were investigated by using pulsed-field gel electrophoresis and probing with sequences cloned from the unstable region which maps near the ends of the linear chromosomal DNA. The chromosomal structures of seven mutant strains harboring large deletions were classified into three types. (i) Deletions internal to one chromosomal arm were characterized in two of the mutant strains. In these strains, a linear chromosomal structure was retained, as were parts of the terminal inverted repeats sequences (TIRs) and the proteins bound to them. (ii) Four of the mutants presented a deletion including all sequences from the TIRs. A junction fragment homologous to sequences originating from internal region of both arms was characterized. Consequently, the chromosomal DNA of these strains was deduced to be circularized. Furthermore, chromosomal stability was assessed in the progeny of these circular DNA mutants. Additional deletion events were detected in 11 mutants among the 13 strains isolated, demonstrating that circular chromosomes do not correspond to a stabilization of the chromosome structure and that the occurrence of deletion could be independent of the presence of chromosomal ends. (iii) A mutant with DNA amplification was shown to have a linear chromosome with a deletion of all sequences between the amplified region and the end of the chromosome. The other chromosomal arm remained unaffected by deletion and associated with protein. PMID- 9226266 TI - Cloning, sequencing, and oxygen regulation of the Rhodobacter capsulatus alpha ketoglutarate dehydrogenase operon. AB - The Rhodobacter capsulatus sucA, sucB, and lpd genes, which encode the alpha ketoglutarate dehydrogenase (E1o), the dihydrolipoamide succinyltransferase (E2o), and the dihydrolipoamide dehydrogenase (E3) components of the alpha ketoglutarate dehydrogenase complex (KGD), respectively, were cloned, sequenced, and used for regulatory analyses. The KGD enzymatic activity was greater in cells grown under aerobic, respiratory growth conditions than under anaerobic, photosynthetic conditions. Similarly, the sucA gene was transcribed differentially, leading to a greater accumulation of sucA mRNAs under respiratory growth conditions than under photosynthetic conditions, although differential rates of mRNA decay could also contribute to the different amounts of sucA mRNAs under these two growth conditions. The sucA promoter was located about 4 kb upstream of the 5' end of the sucA gene, and transcripts greater than 9.5 kb hybridized to a sucA probe, suggesting the presence of an operon that produces a polycistronic mRNA. Thus, these genes seem to be expressed as an unstable primary transcript, and we speculate that posttranscriptional processes control the stoichiometry of KGD proteins. PMID- 9226267 TI - A novel ribosome-associated protein is important for efficient translation in Escherichia coli. AB - Previously, we showed that strains which have been deleted for the 21K gene (hereafter called yfjA), of the trmD operon, encoding a 21-kDa protein (21K protein) have an approximately fivefold-reduced growth rate in rich medium. Here we show that such mutants show an up to sevenfold reduced growth rate in minimal medium, a twofold-lower cell yield-to-carbon source concentration ratio, and a reduced polypeptide chain growth rate of beta-galactosidase. Suppressor mutations that increased the growth rate and translational efficiency of a delta yfjA mutant were localized to the 3' part of rpsM, encoding ribosomal protein S13. The 21K protein was shown to have affinity for free 30S ribosomal subunits but not for 70S ribosomes. Further, the 21K protein seems to contain a KH domain and a KOW motif, both suggested to be involved in binding of RNA. These findings suggest that the 21K protein is essential for a proper function of the ribosome and is involved in the maturation of the ribosomal 30S subunits or in translation initiation. PMID- 9226268 TI - Characterization of mutations affecting the Escherichia coli essential GTPase era that suppress two temperature-sensitive dnaG alleles. AB - Two suppressor mutations of the temperature-sensitive DNA primase mutant dnaG2903 have been characterized. The gene responsible for suppression, era, encodes an essential GTPase of Escherichia coli. One mutation, rnc-15, is an insertion of an IS1 element within the leader region of the rnc operon and causes a polar defect on the downstream genes of the operon. A previously described polar mutation, rnc 40, was also able to suppress dnaG2903. The other mutation, era-1, causes a single amino acid substitution (P17R) in the G1 region of the GTP-binding domain of Era. Analysis of the GTPase activity of the Era-1 mutant protein showed a four to five-fold decrease in the ability to convert GTP to GDP. Thus, lowered expression of wild-type Era caused by the polar mutations and reduced GTPase activity caused by the era-1 mutation suppresses dnaG2903 as well as a second dnaG allele, parB. Phenotypic analysis of the era-1 mutant at 25 degrees C showed that 10% of the cells contain four segregated nucleoids, indicative of a delay in cell division. Possible mechanisms of suppression of dnaG and roles for Era are discussed. PMID- 9226269 TI - Role of the hemA gene product and delta-aminolevulinic acid in regulation of Escherichia coli heme synthesis. AB - We initiated these studies to help clarify the roles of heme, delta aminolevulinic acid (ALA), hemA, and hemM in Escherichia coli heme synthesis. Using recombinant human hemoglobin (rHb1.1) as a tool for increasing E. coli's heme requirements, we demonstrated that heme is a feedback inhibitor of heme synthesis. Cooverexpression of rHb1.1 and the hemA-encoded glutamyl-tRNA (GTR) reductase increased intracellular levels of ALA and heme and increased the rate of rHb1.1 formation. These results support the conclusion that heme synthesis is limited by ALA (S. Hino and A. Ishida, Enzyme 16:42-49, 1973; W. K. Philipp Dormston and M. Doss, Enzyme 16:57-64, 1973) and that the hemA-encoded GTR reductase is a rate-limiting enzyme in the pathway (J.-M. Li, C. S. Russell, and S. D. Cosloy, Gene 82:2099-217, 1989). Increasing the copy number of hemM, whose product is believed to be required for efficient ALA formation (W. Chen, C. S. Russell, Y. Murooka, and S. D. Cosloy, J. Bacteriol. 176:2743-2746, 1994; M. Ikemi, K. Murakami, M. Hashimoto, and Y. Murooka, Gene 121:127-132, 1992), had no effect on either ALA pools or the rate of rHb1.1 accumulation. The hemA-encoded GTR reductase was found to be regulated by ALA. Some of our results differ from those reported by Hart and coworkers (R. A. Hart, P. T. Kallio, and J. E. Bailey, Appl. Environ. Microbiol. 60:2431-2437, 1994), who concluded that ALA formation is not the rate-limiting step in E. coli cells expressing Vitreoscilla hemoglobin. PMID- 9226270 TI - Organization and transcription of the myo-inositol operon, iol, of Bacillus subtilis. AB - Previous determination of the nucleotide sequence of the iol region of the Bacillus subtilis genome allowed us to predict the structure of the iol operon for myo-inositol catabolism, consisting of 10 iol genes (iolA to iouJ); iolG corresponds to idh, encoding myo-inositol 2-dehydrogenase (Idh). Primer extension analysis suggested that an inositol-inducible promoter for the iol operon (iol promoter) might be a promoter-like sequence in the 5' region of iolA, which is probably recognized by sigmaA. S1 nuclease analysis implied that a rho independent terminator-like structure in the 3' region of iolJ might be a terminator for iol transcription. Disruption of the iol promoter prevented synthesis of the iol transcript as well as that of Idh, implying that the iol operon is most probably transcribed as an 11.5-kb mRNA containing the 10 iol genes. Immediately upstream of the iol operon, two genes (iolR and iolS) with divergent orientations to the iol operon were found. Disruption of iolR (but not iolS) caused constitutive synthesis of the iol transcript and Idh, indicating that the iolR gene encodes a transcription-negative regulator (presumably a repressor) for the iol operon. Northern and S1 nuclease analyses revealed that the iolRS genes were cotranscribed from another inositol-inducible promoter, which is probably recognized by sigmaA. The promoter assignments of the iol and iolRS operons were confirmed in vivo with a lacZ fusion integrated into the amyE locus. PMID- 9226271 TI - Suppression of mutants aberrant in light intensity responses of complementary chromatic adaptation. AB - Complementary chromatic adaptation is a process in which cyanobacteria alter the pigment protein (phycocyanin and phycoerythrin) composition of their light harvesting complexes, the phycobilisomes, to help optimize the absorbance of prevalent wavelengths of light in the environment. Several classes of mutants that display aberrant complementary chromatic adaptation have been isolated. One of the mutant classes, designated "blue" or FdB, accumulates high levels of the blue chromoprotein phycocyanin in low-intensity green light, a condition that normally suppresses phycocyanin synthesis. We demonstrate here that the synthesis of the phycocyanin protein and mRNA in the FdB mutants can be suppressed by increasing the intensity of green light. Hence, these mutants have a decreased sensitivity to green light with respect to suppression of phycocyanin synthesis. Although we were unable to complement the blue mutants, we did isolate genes that could suppress the mutant phenotype. These genes, which have been identified previously, encode a histidine kinase sensor and response regulator protein that play key roles in controlling complementary chromatic adaptation. These findings are discussed with respect to the mechanism by which light quality and quantity control the biosynthesis of the phycobilisome. PMID- 9226272 TI - A rubrerythrin operon and nigerythrin gene in Desulfovibrio vulgaris (Hildenborough). AB - Rubrerythrin is a nonheme iron protein of unknown function isolated from Desulfovibrio vulgaris (Hildenborough). We have sequenced a 3.3-kbp Sal1 fragment of D. vulgaris chromosomal DNA containing the rubrerythrin gene, rbr, identified additional open reading frames (ORFs) adjacent to rbr, and shown that these ORFs are part of a transcriptional unit containing rbr. One ORF, designated fur, lies just upstream of rbr and encodes a 128-amino-acid-residue protein which shows homology to Fur (ferric uptake regulatory) proteins from other purple bacteria. The other ORF, designated rdl, lies just downstream of rbr and encodes a 74 residue protein with significant sequence homology to rubredoxins but with a different number and spacing of cysteine residues. Overexpression of rdl in Escherichia coli yielded a protein, Rdl, which has spectroscopic properties and iron content consistent with one Fe3+(SCys)4 site per polypeptide but is clearly distinct from both rubrerythrin and a related protein, nigerythrin. Northern analysis indicated that fur, rbr, and rdl were each present on a transcript of 1.3 kb; i.e., these three genes are cotranscribed. Because D. vulgaris nigerythrin appears to be closely related to rubrerythrin, and its function is also unknown, we cloned and sequenced the gene encoding nigerythrin, ngr. The amino acid sequence of nigerythrin is 33% identical to that of rubrerythrin, and all residues which furnish iron ligands to both the FeS4 and diiron-oxo sites in rubrerythrin are conserved in nigerythrin. Despite the close resemblance of these two proteins, ngr was found to be no closer than 7 kb to rbr on the D. vulgaris chromosome, and Northern analysis showed that, in contrast to rbr, ngr is not cotranscribed with other genes. Possible redox-linked functions for rubrerythrin and nigerythrin in iron homeostasis are proposed. PMID- 9226273 TI - MinCD-independent inhibition of cell division by a protein that fuses MalE to the topological specificity factor MinE. AB - We report that MinE, the topological specificity factor of cell division in Escherichia coli, inhibits septation when fused to the C terminus of the maltose binding protein MalE. This contrasts with overexpression of MinE alone, which affects growth but has no effect on division. Inhibition by MalE-MinE was minCD independent and depended on MinE segments involved in dimerization and prevention of MinCD division inhibition. The SOS and the heat shock responses were not involved, suggesting that the inhibition comes from a direct interaction of MalE MinE with the septation apparatus. MalE-MinE lethality was suppressed by overexpression of ftsZ, as well as by overexpression of ftsN, a suppressor of temperature-sensitive mutations in genes ftsQ, ftsA, and ftsI. We also report that high-level synthesis of MalE disturbs nucleoid partitioning. PMID- 9226274 TI - H-NS and RpoS regulate emergence of Lac Ara+ mutants of Escherichia coli MCS2. AB - Two master growth-phase regulatory proteins, H-NS and sigmaS, are involved in the formation of araB-lacZ fusion clones of Escherichia coli MCS2. The stationary phase sigma factor RpoS is strictly required for the appearance of such mutants, whereas the histone-like protein H-NS represses their emergence. Our results support the idea that genetic changes leading to adaptive mutation in this model system are regulated by physiological signal transduction networks. PMID- 9226275 TI - Evidence for two possible glnB-type genes in Herbaspirillum seropedicae. AB - Two glnB-like genes have been isolated from Herbaspirillum seropedicae by complementation of the Klebsiella pneumoniae glnB502 mutant for growth on nitrate. One of these glnB-like genes has been sequenced and shows strong identity with GlnB proteins derived from other organisms. A Tn5-20 mutation of this glnB was Nif negative. PMID- 9226276 TI - Dual multimodular class A penicillin-binding proteins in Mycobacterium leprae. AB - The ponA gene of cosmid L222 of the Mycobacterium leprae genome library encodes a multimodular class A penicillin-binding protein (PBP), PBP1. The PBP, labelled with a polyhistidine sequence, has been produced in Escherichia coli, extracted from the membranes with 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate (CHAPS) and purified by Ni2(+)-nitrilotriacetic acid-agarose chromatography. In contrast to the pon1-encoded class A PBP1, PBP1 undergoes denaturation at temperatures higher than 25 degrees C, it catalyzes acyl transfer reactions on properly structured thiolesters, and it binds penicillin with high affinity. PMID- 9226277 TI - Identification and characterization of xcpR encoding a subunit of the general secretory pathway necessary for dodecane degradation in Acinetobacter calcoaceticus ADP1. AB - A mutant of Acinetobacter calcoaceticus ADP1 unable to grow on alkanes was complemented for growth on hexadecane with a DNA fragment encoding a protein with homology to XcpR, a subunit of the general secretion pathway for exoproteins in Pseudomonas aeruginosa. Insertional inactivation of xcpR in A. calcoaceticus ADP1 by transcriptional fusion to lacZ abolishes secretion of lipase and esterase and leads to lack of growth on dodecane and slower growth on hexadecane. We, therefore, propose the participation of a secreted protein in alkane degradation. PMID- 9226278 TI - Transposition of the IS21-related element IS1415 in Rhodococcus erythropolis. AB - Three copies of the IS21-related transposable element IS1415 were identified in Rhodococcus erythropolis NI86/21. Adjacent to one of the IS1415 copies, a 47-bp sequence nearly identical to the conserved 5' end of integrons was found. Accurate transposition of IS1415 carrying a chloramphenicol resistance gene (Tn5561) was demonstrated following delivery from a suicide vector to R. erythropolis SQ1. PMID- 9226279 TI - The global regulator CsrA of Escherichia coli is a specific mRNA-binding protein. AB - The csrA gene encodes a global regulatory protein which facilitates glgC mRNA decay in vivo. A purified recombinant CsrA protein was found to inhibit in vitro glg (glycogen biosynthesis) gene expression posttranscriptionally and bind specifically to a glgC runoff transcript without causing its decay. Our results provide further insight into the mechanism by which CsrA functions as an mRNA decay factor. PMID- 9226280 TI - Repair of extensive ionizing-radiation DNA damage at 95 degrees C in the hyperthermophilic archaeon Pyrococcus furiosus. AB - We investigated the capacity of the hyperthermophile Pyrococcus furiosus for DNA repair by measuring survival at high levels of 60Co gamma-irradiation. The P. furiosus 2-Mb chromosome was fragmented into pieces ranging from 500 kb to shorter than 30 kb at a dose of 2,500 Gy and was fully restored upon incubation at 95 degrees C. We suggest that recombination repair could be an extremely active repair mechanism in P. furiosus and that it might be an important determinant of survival of hyperthermophiles at high temperatures. PMID- 9226282 TI - Transcatheter occlusion of patent ductus arteriosus using coil embolization. AB - We studied 31 procedures of coil embolization for occlusion of ductus arteriosus, attempted in 29 patients. The mean age was 4.8+/-3.4 years (1-16 years) and the mean diameter of ductus was 1.8+/-0.7 mm (0.8-3.1 mm). Femoral artery approach was used and aortogram in 90 degrees lateral view was performed. Through a Judkin right coronary catheter, the coil was delivered for occlusion of the ductus. In 5 cases, 2 coils were delivered using retrograde and anterograde techniques. Successful placement of coil was accomplished in 29 procedures. Coils 0.038 inch (diameter)-5 cm (length)-5 mm (helical diameter) (Cook, Inc) were used in 16 procedures, coils 0.035 inch-5 cm-5 mm in 9, coil 0.038 inch-8 cm-8 mm in 1, two coils 0.038 inch-5 cm-5 mm in 2, coils 0.038 inch-5 cm-5 mm+0.038 inch-5 cm-8 mm in 1, and 2 coils 0.035 inch-5 cm-5 mm in 2. Aortogram 20 min after the occlusion, showed residual shunt in 9. Coil migration occurred in a ductus type B in the following day. One patient developed severe haemolysis, due to a change in the coil position, 12 h after the procedure. Echodopplercardiogram 4 to 6 h after the procedure showed a residual shunt in 5 patients, 24 h after in 3 and 30 days after, in 1(3.8%). Heparin therapy started 10 days after occlusion of the ductus, caused reappearance of the shunt in 1 patient. This technique is simple and effective, but complications may occur hours or days after successful ductus occlusion. PMID- 9226281 TI - Abnormal passive head-up tilt test in subjects with symptoms of anxiety power spectral analysis study of heart rate and blood pressure. AB - Previous reports that subjects with anxiety symptoms are at higher risk of sudden death may imply that anxiety induces stable sympathetic hyperactivity. To address this subject, in persons with and without anxiety symptoms, we evaluated autonomic nervous system activity by power spectral analysis of heart-rate and arterial-pressure variability at baseline (rest) and after sympathetic stress (tilt). The 117 subjects selected (56 men and 61 women, age range 23-87 years) were subdivided by questionnaire into three groups: 49 subjects (mean age 55.8+/ 2.8 years) had no anxiety symptoms; 36 (mean age 56.8+/-3.6 years) had one anxiety symptom; and 32 (mean age 55.0+/-2.9 years) had two or more anxiety symptoms. Power spectral analysis recognizes three main components: high frequency (HF), chiefly reflecting vagal efferent activity; low frequency (LF), reflecting sympathetic activity; and very-low-frequency (VLF). The ratio of low- to high-frequency powers (LF:HF) of heart rate variability provides a measure of sympathovagal balance. Power spectral analysis showed that subjects with two or more anxiety symptoms had significantly lower resting values for all power spectral components of heart rate variability: total power (TP), VLF, LF, and HF than did symptomless controls (P<0.05). The highest anxiety-score groups also had a higher baseline LF:HF than the other two groups (P<0.05). Their resting LF:HF ratio correlated positively with anxiety symptom scores (r=0.72, P<0.0001). Tilt induced opposite results: the highest anxiety-score groups had a significantly lower LF:HF ratio; the ratio correlated inversely with their anxiety scores (r= 0.69; P<0.0001). Recordings of resting systolic arterial pressure variability showed that the group with two or more anxiety symptoms had significantly higher LF power (P<0.05) than symptomless controls. Our findings suggest that persons with high anxiety scores have baseline cardiac sympathetic hyperactivity. They also have low heart-rate variability, possibly explaining their susceptibility to sudden cardiac death. PMID- 9226284 TI - Change of complex and periodic heart rate dynamics with change of pulmonary artery pressure in infants with left-to-right shunt lesion. AB - We studied how complex and periodic heart rate dynamic changed as pulmonary artery pressure elevated in 32 infants with ventricular septal defect. In addition, we tested the possibility that the dynamical change could be used to predict the pulmonary artery pressure noninvasively. During cardiac catheterization, mean pulmonary artery pressure was measured and, at the same time, 5-min segments of continuous electrocardiographic recording was stored into computer files. High- (>0.15 hertz) and low- (0.03-0.15 hertz) frequency components of heart rate variability were computed using spectral analysis. The overall complexity of heart rate time series was quantified by its approximate entropy. Pulmonary hypertensive infants (mean pulmonary artery pressure >20 mm Hg, n=17) have significantly lower low- (p<0.05) and high- (p<0.05) frequency power and lower approximate entropy (p<0.0001) than pulmonary normotensive infants (mean pulmonary artery pressure < or =20 mm Hg, n=15). The mean pulmonary artery pressure is significantly correlated not with the spectral powers but with the approximate entropy (r=-0.71, p=0.0001). It can be concluded that, in infants, pulmonary hypertension induced by left- to-right shunt lesions suppresses both periodic and complex heart rate oscillation and that mean pulmonary artery pressure can be predicted by calculating the approximate entropy of heart rate variability. PMID- 9226283 TI - Preoperative management of congestive heart failure in neonates: the closed hood. AB - In this study we report the results of the use of a closed hood with no external administration of CO2 to increase pulmonary vascular resistance by lowering the inspired fraction of oxygen (FiO2) and raising the inspired fraction of carbon dioxide (FiCO2) in patients with congenital heart disease and increased pulmonary blood flow. Between December 1995 and May 1996, 9 neonates (F:5, M:4) were admitted. Each study patient was assigned to clinical classes using a 1 to 4 classification. Ages ranged between 2 and 30 days (mean 18), weight between 2.25 and 3.65 kg (mean 2.89). A plastic hood, closed on the top with a plastic membrane and with the gas entrance open to room air was placed over the head of the patients. Patients increase pCO2 by rebreathing their own expired CO2. After 24 h of the onset of the treatment the media of points of congestive heart failure 1 to 4 classification decrease from a mean of 4 to a mean of 2.28+/-0.44 (p=0.001). A statistically significant improvement in symptoms and lowering of PO2 and pH while raising pCO2 has been demonstrated in this study. PMID- 9226285 TI - Is pregnancy contraindicated after cardiac transplantation? A case report and literature review. AB - We report a cardiac allograft recipient who conceived 5 months after transplantation and spontaneously delivered a full term healthy baby girl. Pregnancy in cardiac transplant recipients is gradually becoming a more frequent issue as more patients in this population consider child bearing. In order to advise patients on potential adverse outcomes due to pregnancy, we reviewed the literature on pregnancy after cardiac transplantation. Published reports show that pregnancy in this population carry a higher risk for complications, in particular there is a higher incidence of pregnancy-induced hypertension, preeclampsia, premature labor, premature and low birth weight infants. The risk for these complications, however, is not higher than for pregnancies of renal or liver transplant recipients, to which pregnancy is not invariably advised against. Despite a greater frequency of complications during pregnancy, successful delivery of a healthy infant is the rule, without any detectable long lasting adverse effects on both mother and offspring. Thus, while cardiac transplant recipients who wish to become pregnant should be counseled on possible complications, it appears that a satisfactory outcome can generally be expected. Additionally, we discuss further issues pertinent to the care of such patients, including hemodynamic changes, immunosuppression, and rejection surveillance during their pregnancies. PMID- 9226286 TI - Heart rate variability after acute myocardial infarction in patients treated with atenolol and metoprolol. AB - Heart rate variability (HRV) reflects autonomous activity that influences the heart. It has been shown that HRV is depressed during acute myocardial infarction (AMI) and that it recovers with time. Beta-blockers reduce mortality after AMI and changes in sympathico-vagal activity have been suggested to be of importance. Under certain animal experimental conditions, metoprolol has been reported to increase vagal tone more than atenolol, which could have clinical implications. The purpose of the present study was to compare the effects of atenolol and metoprolol treatments on HRV during 6 weeks after AMI and to follow the post MI changes in HRV in patients on betablockers. METHODS: In an open, randomised cross over study, 28 patients were randomised to 3+3 weeks' treatment with atenolol or metoprolol starting 2-5 days after AMI. Twenty-four hour Holter recordings were made before randomisation and after 3 and 6 weeks. HRV was analysed as HR, SDRR, SDANN, SD, rMSSD and pNN50 in the time domain and as coefficient of component variance (CCV) of HF and LF, and as LF/HF ratio in the frequency domain. RESULTS: The average daily dose in our study population was 106 mg of metoprolol and 54 mg of atenolol. There were trends toward lower heart rates daytime, lower LF/HF ratio daytime and higher rMSSD on atenolol compared to metoprolol. In the total group of 28 patients we found during the first 3 weeks, a significant increase of SDNN, SDANN (p<0.0001) and LF/HF ratio daytime and CCV-HF night-time (p<0.01). All differences and trends were unchanged between 3 and 6 weeks. CONCLUSIONS: There was no evidence of more increased vagal tone with metoprolol compared to atenolol as has been suggested from animal models. In patients also on chronic treatment with beta blockers, an increase of HRV was seen during the first weeks post MI. PMID- 9226287 TI - Actions and interactions of ethanol and imipramine on the rat sinus node. AB - We studied the actions and interactions of ethanol and imipramine on the sinus node. Strips of the right rat atrium including the sinus node were superfused with Tyrode's solution at 37 degrees C while beating spontaneously. The preparations were exposed to imipramine or ethanol alone as well as to the two drugs in combination while recording membrane potentials with standard intracellular microelectrodes. The results obtained show that ethanol 0.8 and 2.4 g/l exerted a positive chronotropic action. On the other hand, imipramine 0.25 mg/l did not modify the sinus node rate. However, it reduced significantly the positive chronotropic action of ethanol. The sinus node rate decreased under the action of a higher concentration of imipramine (1 mg/l). When ethanol was tested in combination with this concentration of imipramine, the effect of the latter prevailed. In conclusion, a concentration of imipramine that did not affect the sinus node rate antagonized the positive chronotropic action of ethanol. In addition, the negative chronotropic action of a higher concentration of imipramine prevailed over the positive action of ethanol. The results obtained provide additional support to the notion that the use of ethanol and cardioactive drugs in combination may result in significant changes in the actions of either of the two, or both. This is of clinical relevance, since at least some of the individuals under treatment with cardioactive drugs will be alcoholics and/or social drinkers. PMID- 9226288 TI - Radiofrequency ablation of tachyarrhythmias in patients with Ebstein's anomaly. AB - We performed radiofrequency catheter ablation in five patients associated with Ebstein's anomaly to cure their refractory tachyarrhythmias. The presenting arrhythmias were four cases of orthodromic circus movement tachycardia using accessory pathways as a requisite limb, including one case of a Mahaim fiber and one of atrial flutter of common variety. All accessory pathways, including the Mahaim fiber, were ablated by RF energy delivered through the catheter placed at the AV annulus rather than the displaced anatomical AV groove. Interestingly, the antegrade or retrograde conduction interval over these accessory pathways was relatively longer than that of usual accessory pathways, and the accessory pathway potential was fractionated in some cases. The location of the atrioventricular node was displaced from the usual position to the postero inferior area of Koch's triangle in one case. The configuration of the flutter wave was larger than usual in height as well as in width. All tachyarrhythmias were cured by RF catheter ablation. In the case of RF catheter ablation for patients with Ebstein's anomaly, close attention is indispensable in order to accomplish it safely and successfully, because of the anatomical and functional differences peculiar to Ebstein's anomaly. PMID- 9226289 TI - Detection of patients at risk for recurrence of atrial fibrillation after successful electrical cardioversion by signal-averaged P-wave ECG. AB - The aim of the study was to assess the value of signal-averaged ECG of P-wave in predicting recurrence of atrial fibrillation after direct-current electrical cardioversion of chronic atrial fibrillation. The signal-averaged ECG triggered by P-wave was recorded in 35 patients after successful electroconversion. Duration of the high frequency P-wave and the root mean square voltages for the last 20 ms (RMS20) P-wave of the vector magnitude were calculated. After 6 months follow-up recurrence of atrial fibrillation was observed in 11 patients (group I) and in 24 patients sinus rhythm was maintained (group II). A filtered P-wave was significantly longer in group I with recurrence of atrial fibrillation, than in patients from group II who maintained sinus rhythm (145+/-11.8 vs 130+/-10.8 ms, p<0.001). RMS20 was significantly lower in group I than in patients from group II (1.6+/-0.6 vs 2.2+/-0.9 microV, p<0.02). A filtered P-wave of duration >q37 ms associated with a RMS 20 ms <1.9 microV had a sensitivity of 73% and specificity of 71% for the detection of patients with recurrence of atrial fibrillation after successful direct-current electrical cardioversion of chronic atrial fibrillation. These results suggest that signal-averaged ECG of P-wave may be helpful for identification of patients with recurrence of atrial fibrillation after successful direct-current electrical cardioversion. PMID- 9226290 TI - Clinical course of Chagas' heart disease: a comparison with dilated cardiomyopathy. AB - This investigation was carried out to compare the clinical course of patients with chronic Chagas' heart disease with that of patients with dilated cardiomyopathy. A total of 125 patients (75 chagasic and 50 nonchagasic) prospectively followed up at the Cardiomyopathy clinic of Santa Casa Hospital from January 1990 to June 1993 entered the study. Patients underwent clinical history, physical examination, serological tests, resting electrocardiogram, chest X-ray and two-dimensional echocardiography. In nonchagasic patients, hypertensive cardiomyopathy was found in 17 of 50 (34%) patients, idiopathic dilated cardiomyopathy in 16 (32%), the association of hypertension and coronary artery disease in 12 (24%) and ischemic cardiomyopathy in two (4%). Twenty-one (23%) chagasic and three (6%) nonchagasic patients died during the study period (P = 0.02). Sudden cardiac death occurred in eight (38%) chagasic patients, pump failure death was detected in 10 (47%) and the mode of death could not be determined in three (14%) patients with chronic Chagas' heart disease. Thus, patients with chronic Chagas' heart disease have a clinical course worse than that of patients with nonchagasic dilated cardiomyopathy. This fact may be ascribed to the electrocardiographic and morphological peculiarities usually found in chronic Chagas' heart disease. PMID- 9226291 TI - Impact of various compression rates on interpretation of digital coronary angiograms. AB - According to the ACC/ACR/NEMA/ESC-guidelines, digital techniques should be replaced by cinefilm for coronary angiography. The ad hoc group of experts recently chose CD-R (CD recordable) as transport media and the JPEG standard for image compression. To avoid a possible loss of image quality, the guidelines allow a maximal data compression of only 2:1. This, however, leads to a considerable limitation: coronary angiograms cannot be viewed in real-time directly from CD. Since the possible influence of higher compression rates on image quality of coronary angiograms had not been investigated in a controlled study, we evaluated 8 various compression rates (ranging from 5:1 to 43:1) according to a prospective, randomized and blinded protocol. Four independent observers assessed 1440 angiograms using a semiquantitative score. We found that angiograms with a compression rate of 5:1 and 6:1 did not lead to a clinically relevant deterioration of image quality, whereas 11:1 was still acceptable, but 43:1 becomes unacceptable. Since no clinically relevant loss of information at a compression rate of 6:1 was experienced in our study, a modification of the ACC/ACJ/NEMA/ESC-guidelines allowing higher compression rates should be considered. PMID- 9226292 TI - Restenosis or rapid progression in non-dilated sites are not predictors of late spontaneous coronary events. AB - The present study was designed to assess the prognostic value of clinical and angiographic factors, and especially restenosis or rapid progression in non dilated sites, on major spontaneous coronary events at long-term follow-up after a first successful coronary angioplasty performed for angina pectoris. A second aim was to assess the prognostic factors and especially restenosis in asymptomatic patients after angioplasty. The first 352 consecutive patients undergoing a successful coronary angioplasty were selected and followed-up. The following variables: age, sex, unstable angina, previous myocardial infarction, diabetes, hypercholesterolemia, tobacco consumption, hypertension, fibrinogen, coronary extent, single or multiple dilatation, restenosis, new progression, clinical deterioration of anginal status just before angiographic restudy or asymptomatic status were subjected to a stepwise regression analysis. Restenosis (a loss of 30% in diameter and/or a return to a >50% stenosis) and progression in non-dilated segments (a 20% reduction in diameter) were assessed by a computer assisted method. Cardiac death, new myocardial infarction or new unstable angina, at long-term follow-up after angiographic restudy, were regarded as spontaneous coronary events and pooled in a single dependent variable. Thus 41 patients had a coronary event. In the overall population, clinical deterioration of anginal status (p<0.001, relative risk: 3.65) and fibrinogen (p<0.05, relative risk: 1.03) were independent predictors of spontaneous coronary events. Restenosis or new progression were not predictors. In asymptomatic patients (n=187), fibrinogen (p<0.01, relative risk=1.06) was the only predictor and restenosis was not an independent predictor of spontaneous coronary events. The best predictor of spontaneous coronary events at long-term follow-up after a first successful coronary angioplasty is clinical deterioration in anginal status in the months following the procedure. Restenosis does not appear as an independent predictor. Rapid progression observed in non-dilated sites is not an important prognostic factor. PMID- 9226293 TI - Isolated mitral regurgitation complicating relapsing polychondritis. AB - The reported incidence of mitral regurgitation in relapsing polychondritis ranges from 2 to 3% and is associated with aortic regurgitation. There are no reports that mitral regurgitation can be an isolated cardiac complication of relapsing polychondritis. This case report demonstrates that partial chordal rupture and consequent severe mitral regurgitation can be the only features of cardiac involvement in relapsing polychondritis. PMID- 9226294 TI - Mitral valve prolapse secondary to rheumatic valvulitis. PMID- 9226295 TI - Sudden hemorrhagic tamponade simulating subacute ventricular rupture after acute myocardial infarction. PMID- 9226296 TI - A Symposium on Signal Transduction in the Endothelium. Vancouver, September 9-11, 1996. PMID- 9226297 TI - Signal transduction mechanisms mediating the vascular actions of endothelin. AB - Endothelin (ET)-1, an endothelium-derived vasoactive polypeptide encoded in the human genome, is the most potent vasoconstrictor identified to date. In addition to its acute role in modulating vascular smooth muscle tone, ET-1 also plays a critical role in the long-term control of cellular growth within the vasculature and thus, modulates the chronic remodeling of the vascular tree. In order to produce such a diverse range of biological responses, this peptide is able to activate numerous distinct effector systems including phospholipase C, phospholipase D, phospholipase A2, adenylate and guanylate cyclases and numerous cytosolic/nuclear protein kinases. These actions, mediated via an interaction with two major subtypes of cell surface seven-transmembrane receptors (ET(A) and ET(B)), are coupled to their effector systems by several distinct types of guanine nucleotide regulatory proteins (both inhibitory and stimulatory G proteins). This review describes such intercations and how distinct pharmacological agents have been used to identify the diverse signaling mechanisms utilized by the ET isopeptides. PMID- 9226299 TI - G proteins and endothelium-dependent relaxations. AB - Endothelial cells control the tone of the underlying smooth muscle by releasing relaxing factors (nitric oxide, NO, prostacyclin and endothelium-derived hyperpolarizing factor). G proteins couple a number of endothelial cell receptors to the activation of NO synthase. Pertussis toxin selectively ADP-ribosylates certain G proteins (mainly G(i)). In the porcine coronary artery, pertussis toxin inhibits the release of NO evoked by certain (serotonin, alpha2-adrenergic agonists, leukotrienes, thrombin), but not all, (bradykinin, adenosine diphosphate) endothelium-dependent vasodilators. This suggests that both G(i) and G(q) proteins can couple receptor activation to the increase in endothelial Ca2+ concentration required to stimulate NO synthase. In arteries with regenerated endothelium and in cultured endothelial cells, the release of NO evoked by pertussis-toxin-sensitive mechanisms is severely reduced or absent, while the response to other endothelium-dependent agonists is normal. To judge from experiments with cultured endothelial cells, the curtailment in pertussis-toxin sensitive release of NO is due to an abnormal function rather than a reduced presence of G(i) proteins, or a reduced sensitivity of the cell membrane receptor. The selective impairment of G(i) proteins in regenerated endothelial cells predisposes the blood vessel wall to vasospasm and to the initiation of the atherosclerotic process. PMID- 9226298 TI - Calcium-dependent and calcium-independent activation of the endothelial NO synthase. AB - Largely assumed to be a Ca2(+)-/calmodulin-dependent enzyme, the endothelial constitutive nitric oxide (NO) synthase (NOS III) can be activated by agonists as a consequence of an increase in the intracellular concentration of free Ca2+ ([Ca2+]i). This increase in [Ca2+]i is elicited by an increase in inositol 1,4,5 trisphosphate which is the consequence of tyrosine phosphorylation and activation of phospholipase C-gamma1 as well as protein tyrosine phosphatases. Following the mobilization of intracellular Ca2+, the depleted Ca2+ stores signal to cation channels in the plasma membrane by a pathway which appears to involve activation of both tyrosine and serine/threonine kinases since this portion of the Ca2+ response is attenuated by both tyrosine kinase inhibitors and serine phosphatase inhibitors. In response to fluid shear stress the continuous production of NO by native and cultured endothelial cells is associated with only a transient and minimal increase in [Ca2+]i. In the absence of extracellular Ca2+ and in the presence of the calmodulin antagonist, shear stress stimulates a continuous production of NO which is sensitive to the nonspecific kinase inhibitor staurosporine and the tyrosine kinase inhibitor erbstatin A. A pharmacologically identical activation of NOS III can be induced by protein phosphatase inhibitors suggesting that the tyrosine phosphorylation of NOS III or an associated regulatory protein is crucial for its Ca2(+)-independent activation. Thus in a departure from widely held beliefs, we propose that the endothelial cells are able to respond to mechanical and humoral stimuli activating NOS III by at least two separate pathways. PMID- 9226300 TI - Calcium-release-activated calcium influx in endothelium. AB - Signaling pathways activated by the tachykinin substance P (SP) were investigated in pig coronary artery endothelial cells (PCAECs). Single cells were obtained after enzymatic digestion of coronary arteries. Intracellular Ca2+ ([Ca2+]i) was measured from fura-2 fluorescence while membrane potential or ionic current was measured using patch-clamp techniques. In physiological saline solution, SP induced hyperpolarizations or outward currents which coincided with biphasic [Ca2+]i increases representing store release of Ca2+ and Ca2+ influx. Single channel recording protocols showed that both sources of Ca2+ activated a small conductance K+ channel, resulting in cell hyperpolarization. When outward currents were blocked by d-tubocurare, Cs+, or BAPTA, an inward current was unmasked. Ion substitution protocols showed that the SP-induced inward current was (1) carried by a mixture of Ca2+ and Na+, (2) blocked by La3+, and (3) inactivated by high extracellular [Ca2+]. Tyrosine kinase inhibitors also blocked the inward current. The same current was activated by bath application of BHQ, an inhibitor of the endoplasmic reticulum Ca2+ ATPase, or by cell dialysis with IP3. These results suggest that the plateau phase of the agonist-activated [Ca2+]i increase in PCAECs reflects Ca2+ entry through a depletion-activated Ca2+ channel. The characteristics of this channel are compared to those of Ca2+ channels found in other nonexcitable cells. PMID- 9226301 TI - Multiple mechanisms of activating Ca2+ entry in freshly isolated rabbit aortic endothelial cells. AB - In Fura-2-loaded, freshly isolated rabbit aortic endothelial cells the Ca2+ entry pathway was investigated using the Mn2(+)-quenching technique. Acetylcholine (ACh) interaction with muscarinic receptors activated Mn2+ influx through the plasma membrane. Sarcoplasmic-endoplasmic reticulum Ca2+ ATPase blockers such as cyclopiazonic acid (CPA), thapsigargin and BHQ, which block the endoplasmic reticulum Ca2+ pump and do not interact with receptors, also activated Mn2+ influx. Mn2+ influx activated by either ACh or CPA was blocked by the following agents: SKF96365, a receptor-operated Ca2+ channel (ROC) blocker; NCDC, a PLC and ROC blocker, and genistein, a tyrosine kinase inhibitor. D600, the L-type Ca2+ channel blocker, had no significant effect on Mn2+ influx. Caffeine blocked the ACh-induced Ca2+ release but had no effect on the ACh-induced Mn2+ influx. Similarly dantrolene, which blocked intracellular Ca2+ release induced by ACh, did not affect the ACh-activated Mn2+ influx. These data suggest that ACh can activate Ca2+ influx without depletion of the ACh-sensitive intracellular Ca2+ store. It is concluded (1) that in freshly isolated endothelial cells depletion of the intracellular Ca2+ store is not necessary for ACh-activated Ca2+ influx, and (2) that receptor activation and intracellular Ca2+ store depletion may activate the same Ca2+ entry pathway through parallel mechanisms. PMID- 9226302 TI - Overview: temporal and spatial relationships in shear stress-mediated endothelial signalling. PMID- 9226303 TI - Mechanotransduction in endothelial cells: temporal signaling events in response to shear stress. AB - Fluid shear stress is one of the most important mechanical forces acting upon vascular endothelium, because of its location at the interface between the bloodstream and vascular wall. Recent evidence indicates that several intracellular signaling events are stimulated in endothelial cells in response to shear stress. Through these events, shear stress modulates endothelial cell function and vascular structure, but the molecular basis of shear stress mechanotransduction remains to be elucidated. In our research we have focused on three temporal signal responses to shear stress: (1) production of nitric oxide (NO) as an immediate response; (2) activation of extracellular-regulated kinases (ERK1/2; p44/p42 mitogen-activated protein (MAP) kinases) as a rapid response, and (3) tyrosine phosphorylation of focal adhesion kinase (FAK) as a sustained response. In terms of vessel biology, NO production, and ERK1/2 and FAK activation seem to be correlated with vascular homeostasis, gene expression and cytoskeletal rearrangement, respectively. In this review, we discuss the mechanisms that establish the temporal order of shear stress-stimulated responses based on a hierarchy for assembly of signal transduction molecules at the cell plasma membrane. PMID- 9226304 TI - Multiple types of chloride channels in bovine pulmonary artery endothelial cells. AB - We have characterized two different types of Cl- currents in calf pulmonary artery endothelial (CPAE) cells by using a combined patch-clamp and Fura-2 microfluorescence technique to measure simultaneously ionic currents and the intracellular Ca2+ concentration, [Ca2+]i. Exposure of CPAE cells to 28% hypotonic solution induces cell swelling without a change in membrane capacitance and [Ca2+]i, and concomitantly activates a current. This current, I(Cl, vol), is closely correlated with the changes in cell volume and shows a modest outward rectification. It slowly inactivates at potentials more positive than +60 mV but is time- and voltage-independent at other potentials. Increase in [Ca2+]i by different maneuvers, such as application of vasoactive agonists (ATP), ionomycin, or loading of the cells directly with Ca2+ also activates a Cl- current, I(Cl, Ca). This current slowly activates at positive potentials, inactivates quickly at negative potentials and shows strong outward rectification. A time-independent component of the current activated by elevation of [Ca2+]i alone can be inhibited by cell shrinking by exposing the cells to hypertonic solution, indicating that an increase in [Ca2+]i also co-activates I(Cl, vol). Forskolin or cAMP never activated a current in CPAE cells, which indicates the lack of cAMP-activated channels in these cells. There is also no evidence for the existence of voltage gated Cl- channels in resting, nonstimulated cells. Challenging a cell with elevated [Ca2+]i and hypotonic solutions activated I(Cl, vol) on top of I(Cl, Ca), suggesting that I(Cl, Ca) and I(Cl, vol) are different channels. We conclude that CPAE cells do not express voltage-gated (ClC-type) or cAMP-gated (CFTR-type) Cl- channels, but activate large Cl- currents after volume (mechanical?) or chemical (Ca2+) stimulation. PMID- 9226305 TI - Potential cellular signaling mechanisms mediating upregulation of endothelial nitric oxide production by estrogen. AB - Experimental and clinical studies have provided ample evidence that estrogens exert a significant antiatherosclerotic effect and reduce morbidity and mortality due to cardiovascular diseases. The exact cellular mechanism of this vasculoprotective action of estrogen is not known, but recent work in our and other laboratories suggests that upregulation of endothelial nitric oxide (NO) production may significantly contribute to the mechanism. The vascular endothelium of female animals and humans produces more NO than that of males. Estrogen treatment significantly increases endothelial NO generation in ovariectomized animals and in postmenopausal women. Reduced endothelial NO production in the aorta of estrogen-receptor-deficient mice indicates that the nuclear estrogen receptor mediates the effect of estrogen. The most probable mechanism of estrogen-induced upregulation of endothelial NO production is the transcriptional stimulation of NOS III gene expression. However, the following alternative mechanisms may be involved as well: (1) inhibition of cytokine induced downregulation of NOS III gene expression, (2) posttranslational modification of NOS III protein, (3) increased cofactor or L-arginine availability, (4) nongenomic activation of second messengers (e.g., Ca2+, cAMP) and tyrosine kinase, and (5) modulation of NO degrading systems (e.g., reactive oxygen radical generation and antioxidants). This paper reviews current data supporting these potential mechanisms. PMID- 9226306 TI - Cytochrome P450 metabolites of arachidonic acid as intracellular signaling molecules in vascular tissue. AB - Recent studies from our laboratory have indicated that vascular smooth muscle cells (VSMC) metabolize arachidonic acid via a P4504A-dependent pathway to 20 hydroxyeicosatetraenoic acid (20-HETE), and that this system serves as a novel signal transduction pathway that plays a central role in the regulation of vascular tone. The major metabolite of arachidonic acid formed in cerebral and renal arteries is 20-HETE. The mRNA and protein for P4504A enzymes, which produce 20-HETE, have been localized in VSMC. 20-HETE is a potent vasoconstrictor, that acts in part by inhibition of the opening of the large conductance, calcium activated potassium channel, and depolarizes VSMC membrane. A preliminary study also indicated that 20-HETE activates the L-type calcium current in cerebral arterial smooth muscle. Inhibition of the endogenous production of 20-HETE in renal and cerebral arterioles attenuates pressure-dependent myogenic tone in vitro, as well as autoregulation of renal and cerebral blood flow in vivo. There is also evidence that indicates that nitric oxide regulates the formation of 20 HETE by binding and inactivating the P450 heme moiety, thus providing a negative feedback control mechanism for this system. The data outlined suggest that 20 HETE could act as a intracellular second messenger that plays an integral role in the signal transduction processes underlying the development of pressure dependent myogenic tone. PMID- 9226307 TI - Why is female sex an independent predictor of shortened overall survival after proton/photon radiation therapy for skull base chordomas? PMID- 9226308 TI - Neuropsychological function in adults after high dose fractionated radiation therapy of skull base tumors. AB - PURPOSE: To evaluate the long term effects of high dose fractionated radiation therapy on brain functioning prospectively in adults without primary brain tumors. METHODS AND MATERIALS: Seventeen patients with histologically confirmed chordomas and low grade chondrosarcomas of the skull base were evaluated with neuropsychological measures of intelligence, language, memory, attention, motor function and mood following surgical resection/biopsy of the tumor prior to irradiation, and then at about 6 months, 2 years and 4 years following completion of treatment. None received chemotherapy. RESULTS: In the patients without tumor recurrence or radiation necrosis, there were no indications of adverse effects on cognitive functioning in the post-acute through the late stages after brain irradiation. Even in patients who received doses of radiation up to 66 Cobalt Gy equivalent through nondiseased (temporal lobe) brain tissue, memory and cognitive functioning remained stable for up to 5 years after treatment. A mild decline in psychomotor speed was seen in more than half of the patients, and motor slowing was related to higher radiation doses in midline and temporal lobe brain structures. CONCLUSION: Results suggest that in adults, tolerance for focused radiation is relatively high in cortical brain structures. PMID- 9226309 TI - Large intracranial vessel occlusive vasculopathy after radiation therapy in children: clinical features and usefulness of magnetic resonance imaging. AB - PURPOSE: To assess the relationship between large intracranial vessel occlusive vasculopathy (vasculopathy) and radiation therapy, and to clarify the clinical efficacy of magnetic resonance (MR) imaging in the diagnosis and screening of the vasculopathy. METHODS AND MATERIALS: We retrospectively evaluated the medical records and serial MR images for 32 pediatric patients, in whom radiation therapy had been given to fields including the circle of Willis and major cerebral arteries. All children had periodically undergone follow-up neurologic assessment and MR imaging examinations at Kanagawa Children's Medical Center for more than one year after radiation therapy (range 1.3-14 years). Patients who had not remained free of tumor progression up to the time of final evaluation were excluded. RESULTS: Vasculopathy developed in 6 of 32 patients 2-13 years after radiation therapy. Three of them presented with transient ischemic attacks (TIA) and the other three showed infarctions without preceding TIA. Steno-occlusive changes of major cerebral arteries were identified by MR imaging in all six patients, but not obtained in the remaining 26 patients. In the patients with TIA, MR imaging demonstrated steno-occlusive changes at the time of TIA, before irreversible infarction. They have been doing well subsequent to encephaloduroarteriosynangiosis. In the three patients who presented infarction without preceding TIA, MR imaging did not demonstrate the vascular change before the onset of infarction, and two of them developed neurologic deficits. The mean exposure dose for the circle of Willis and major cerebral arteries in these six patients was significantly higher than that in the remaining 26 patients without this sequela (61 Gy vs. 50 Gy, p < 0.05). The mean age at radiation therapy of the six patients was lower, but the difference was not significant. CONCLUSION: The incidence of vasculopathy after radiation therapy has a considerable correlation with radiation dose and age at radiation therapy. MR examination is useful for the diagnostic evaluation of vasculopathy, and it is also effective in screening for vasculopathy in patients with TIA, and may be helpful in the prevention of neurologic sequela. PMID- 9226310 TI - External beam irradiation for choroid metastases: identification of factors predisposing to long-term sequelae. AB - PURPOSE: To improve overall quality of life, palliative treatments should attempt to minimize associated complications while effectively controlling specific symptoms. We reviewed our experience treating posterior uveal metastases with external beam radiotherapy (EBRT) to determine the complication rate and to identify the relationship between patient, tumor, or treatment-related factors and the development of ocular complications. METHODS AND MATERIALS: 483 consecutive patients (pts) (578 eyes) were diagnosed with intraocular metastatic disease from solid tumors between 1972-1995. Of these, 233 eyes (188 pts) had lesions of the posterior uveal tract and received EBRT. Median follow-up time was 5.8 months (range: 0.7-170.0 months). Follow-up information regarding the development of complications was documented for 230 eyes. Complete EBRT details were available for 189 eyes. Seventy-two percent of the patients received 30.0 40.0 Gy in 2.0-3.0 Gy fractions. Biologically effective dose (BED) was calculated to allow meaningful comparisons between various fractionation regimens and total doses. Concurrent chemotherapy and/or hormonal therapy was used for 101 eyes (44%). RESULTS: Median BED was 61 Gy3 (range, 6.7-105 Gy3), and 80% of treated eyes received BED 50-70 Gy3. EBRT energies included photons (70%), 60Co (19%), electrons (6%), mixed energies (3%), and orthovoltage (2%). Lens-sparing techniques were used in 136 eyes (71%). At last follow-up 28 eyes (12%) developed one or more significant complications, including cataracts (16 eyes), radiation retinopathy (6 eyes), optic neuropathy (5 eyes), exposure keratopathy (5 eyes), and neovascularization of the iris (4 eyes). Two eyes developed narrow-angle glaucoma, and one of these required enucleation. On univariate analysis, Caucasian race (vs. Black/Hispanic, p = 0.03), increased intraocular pressure at diagnosis (>21 mmHg, p = 0.02), and diagnosis by biopsy (vs. no biopsy, p = 0.03) predisposed toward the development of complications. Factors not correlated with complications included BED (p = 0.18), energy type (p = 0.81), lens-sparing technique (versus whole globe, p = 0.57), and concurrent systemic treatment (p = 0.60). The small number of complications did not support a multivariate analysis. CONCLUSIONS: Despite the employment of a variety of EBRT treatment techniques and the proximity of choroidal metastases to radiosensitive structures, significant complications of palliative EBRT were infrequent. Although complications do occur, they are related to host factors and do not appear to be a function of irradiation parameters. We conclude that the potential benefits of vision and globe preservation after palliative EBRT outweigh the small risk of treatment induced complications. PMID- 9226311 TI - Neon heavy charged particle radiotherapy of glioblastoma of the brain. AB - PURPOSE: High-linear energy transfer (LET) radiation beams have potential applications in the treatment of glioblastoma, but have not yet demonstrated significant improvement in results. However, some patients have had local control of glioblastoma with high-LET irradiations such as neutrons and heavy charged particles. METHODS AND MATERIALS: In this collaborative study, 15 patients were entered into a randomized protocol comparing two dose levels of 20 and 25 Gy in 4 weeks of neon ion irradiation. This trial was intended to determine the optimal neon dose in terms of survival and effects of radiation. RESULTS: Fourteen patients were evaluable with no significant differences in median survival (13 and 14 months; p = NS) or median time to failure (7 and 9 months; p = NS) between the two dose arms. Three patients died of nontumor-related causes, of whom one (who died 19 months posttreatment) had autopsy confirmation of no tumor on pathological exam. The other two patients had stable magnetic resonance imaging scans at 6 and 22 months posttreatment. CONCLUSION: Although the results did not demonstrate the optimal high-LET dose level, there is an intriguing effect in that two patients had control of glioblastoma until death at 19 and 22 months. This suggests that better conformation of the high-LET dose to the tumor with neutron capture therapy or dynamic conformal heavy charged particle therapy might control glioblastoma while minimizing brain damage from radiation. PMID- 9226312 TI - Efficacy of targeted supradose cisplatin and concomitant radiation therapy for advanced head and neck cancer: the Memphis experience. AB - PURPOSE/OBJECTIVE: To evaluate the feasibility, response rates, and toxicity of a Phase II study using targeted supradose cisplatin and concurrent radiation therapy in unresectable Stage III-IV head and neck squamous cell carcinoma. METHODS AND MATERIALS: Sixty patients presenting between 6/93-9/94 were enrolled, 44 (73%) of whom had T4 and/or N2-N3 nodal disease. All patients were treated with rapid targeted superselective intraarterial infusions of cisplatin (150 mg/m2 weekly x 4) and simultaneous sodium thiosulfate intravenously (9 g/m2) for systemic neutralization of cisplatin. Concurrent (day 1) daily radiation therapy was delivered to the primary tumor and overt nodal disease to 66-74 Gy while the uninvolved lower neck received 50 Gy, at 2.0 Gy/fraction. RESULTS: Fifty-one (85%) patients completed the full RADPLAT protocol as planned. Fifty-seven of 60 patients were evaluable for response. Histological (n = 50) or clinical (n = 7) assessment of primary site revealed a complete response (CR) in 52 patients, partial response (PR) in 4, and stable disease (SD) in 1. Of the 40 patients presenting with nodal metastases, pathological (n = 31) or clinical (n = 6) assessment revealed a CR in 25, PR in 11, and SD in 1, while 3 were unevaluable. Overall, for both primary site and nodal disease, CR was attained in 44 (75%), PR in 12 (23%), and SD in 1 (2%) of the 57 evaluable patients. Only 2 (4%) of 57 evaluable patients have recurred above the clavicle, 1 in the primary site and 1 in the regional lymph nodes. Twelve patients (23%) have failed in distant sites. Grade III/VI toxicity has included gastrointestinal in 6, hematologic in 6, mucosal in 12, vascular in 4, and neurological in 4 patients. CONCLUSION: Concurrent radiation therapy and targeted supradose cisplatin (i.e., RADPLAT) can be safely delivered with high response rates and excellent loco-regional control in advanced Stage III/IV head and neck squamous cell carcinoma. PMID- 9226313 TI - Dose-volume complication analysis for visual pathway structures of patients with advanced paranasal sinus tumors. AB - PURPOSE: The purpose of the present work was to relate dose and volume information to complication data for visual pathway structures in patients with advanced paranasal sinus tumors. METHODS AND MATERIALS: Three-dimensional (3D) dose distributions for chiasm, optic nerve, and retina were calculated and analyzed for 20 patients with advanced paranasal sinus malignant tumors. 3D treatment planning with beam's eye view capability was used to design beam and block arrangements, striving to spare the contralateral orbit (to lessen the chance of unilateral blindness) and frequently the ipsilateral orbit (to help prevent bilateral blindness). Point doses, dose-volume histogram analysis, and normal tissue complication probability (NTCP) calculations were performed. Published tolerance doses that indicate significant risk of complications were used as guidelines for analysis of the 3D dose distributions. RESULTS: Point doses, percent volume exceeding a specified published tolerance dose, and NTCP calculations are given in detail for patients with complications versus patients without complications. Two optic nerves receiving maximum doses below the published tolerance dose sustained damage (mild vision loss). Three patients (of 13) without optic nerve sparing and/or chiasm sparing had moderate or severe vision loss. Complication data, including individual patient analysis to estimate overall risk for loss of vision, are given. CONCLUSION: 3D treatment planning techniques were used successfully to provide bilateral sparing of the globe for most patients. It was more difficult to spare the optic nerves, especially on the ipsilateral side, when prescription dose exceeded the normal tissue tolerance doses. NTCP calculations may be useful in assessing complication risk better than point dose tolerance criteria for the chiasm, optic nerve, and retina. It is important to assess the overall risk of blindness for the patient in addition to the risk for individual visual pathway structures. PMID- 9226314 TI - Tumor hypoxia adversely affects the prognosis of carcinoma of the head and neck. AB - PURPOSE: Tumor hypoxia adversely affects short term clinical radiation response of head and neck cancer lymph node metastases and long term disease-free survival (DFS) in cervix carcinoma. This study was performed to evaluate the relationship between tumor hypoxia and DFS in patients with squamous carcinoma of the head and neck (SCCHN). METHODS AND MATERIALS: Pretreatment tumor pO2 was assessed polarographically in SCCHN patients. All patients were AJCC Stage IV and had pretreatment oxygen measurements taken from locally advanced primaries (T3 or T4) or neck nodes > or = 1.5 cm diameter. Treatment consisted of once daily (2 Gy/day to 66-70 Gy) or twice daily irradiation (1.25 Gy B.I.D. to 70-75 Gy) +/- planned neck dissection (for > or = N2A disease) according to institutional treatment protocols. RESULTS: Twenty-eight patients underwent tumor pO2 measurement. The average pre-treatment median pO2 was 11.2 mm Hg (range 0.4-60 mm Hg). The DFS at 12 months was 42%. The DFS was 78% for patients with median tumor pO2 > 10 mm Hg but only 22% for median pO2 < 10 mm Hg (p = 0.009). The average tumor median pO2 for relapsing patients was 4.1 mm Hg and 17.1 mm Hg in non-relapsing (NED) patients (p = 0.007). CONCLUSION: Tumor hypoxia adversely affected the prognosis of patients in this study. Understanding of the mechanistic relationship between hypoxia and treatment outcome will allow for the development of new and rational treatment programs in the future. PMID- 9226315 TI - The value of breast lumpectomy margin assessment as a predictor of residual tumor burden. AB - PURPOSE: Margin assessment is commonly used as a guide to the relative aggressiveness of therapy for breast conserving treatment (BCT), though its value as a predictor of the presence, type, or extent of residual tumor has not been conclusively studied. Controversy continues to exist as to what constitutes a margin that is "positive," "close," or "negative." We attempt to address these issues through an analysis of re-excision specimens. PATIENTS AND METHODS: As part of an institutional prospective practice approach for BCT, 265 cases with AJCC Stage I/II carcinoma with an initial excision margin that was < or = 2 mm or indeterminate were subjected to re-excision. The probability of residual tumor (+RE) was evaluated with respect to tumor size, histopathologic subtype, relative closeness of the measured margin, the extent of margin positivity graded as focal, minimal, moderate, or extensive, and the extent of specimen processing as reflected in the number of cut sections per specimen volume (S:V ratio). The amount of residual tumor was graded as microscopic, small, medium, or large. The histopathologic subtype of tumor in the re-excision specimen was classified as having an invasive component (ICa) or pure DCIS (DCIS). RESULTS: The primary excision margin was positive, > 0 < or = 1 mm, 1.1-2 mm, and indeterminate in 60%, 18%, 5%, and 17%, respectively. The predominant histopathologies in the initial excision specimens were invasive ductal (IDC) (50%) and tumors with an extensive intraductal component (EIC) (43%). The histopathology of the initial excision specimen was highly predictive of the histopathology of tumor found on re-excision, as residual DCIS was found in 60% of +RE specimens with initial histopathology of EIC compared to 26% for IDC (p = 0.001). Neither the extent of margin positivity nor the extent of tumor in the re-excision were significantly related to the initial histopathologic subtype; however, a +RE was seen in 59% of EIC, 43% of IDC, and 32% of invasive lobular ILC cases (p = 0.01). The extent of margin positivity was significantly related to the size of the tumor such that tumor size < or = 20 mm was associated with a greater probability of focal or minimal margin involvement. Positive margins graded as focal, minimal, moderate/extensive were associated with a +RE in 26%, 58%, and 84%, respectively (p = 0.001). Further, the extent of positivity was significantly correlated with the extent of residual tumor such that focal/minimal positivity was more commonly associated with micro/small +RE, whereas moderate/extensive positivity was associated with medium/large +RE. When the closest margin of the initial excision specimen was positive, > 0 < or = 1 mm, or 1.1-2 mm, a +RE was found in 56%, 41%, and 17%, respectively (p = 0.01) but did not correlate with the amount of residual tumor. The extent of specimen processing as reflected in the S:V ratio did not correlate with the probability of defining a measured margin as positive nor the probability of a +RE. In a univariate model, the extent of tumor in the re-excision and the histologic type of tumor in the re-excision were significantly associated with margin status and initial histopathology, respectively. The probability of finding a +RE, based on a multivariate model, was associated with the closeness and extent of margin involvement and initial histopathology of an EIC. CONCLUSION: The relative closeness of tumor to the specimen edge and the extent of margin positivity are predictive for residual tumor, though with an error consistent with its limitations as a sampling procedure. The histopathology of tumor in the initial excision is predictive of the type of residual tumor and the extent of margin positivity was correlated with the amount of residual tumor. PMID- 9226316 TI - Low-dose-rate brachytherapy as the sole radiation modality in the management of patients with early-stage breast cancer treated with breast-conserving therapy: preliminary results of a pilot trial. AB - PURPOSE: We present the preliminary findings of our in-house protocol treating the tumor bed alone after lumpectomy with low-dose-rate (LDR) interstitial brachytherapy in selected patients with early-stage breast cancer treated with breast-conserving therapy (BCT). METHODS AND MATERIALS: Since March 1, 1993, 60 women with early-stage breast cancer were entered into a protocol of tumor bed irradiation only using an interstitial LDR implant with iodine-125. Patients were eligible if the tumor was < or = 3 cm, margins were > or = 2 mm, there was no extensive intraductal component, the axilla was surgically staged, and a postoperative mammogram was performed. Implants were placed using a standardized template either at the time of reexcision or shortly after lumpectomy. A total of 50 Gy was delivered at 0.52 Gy/h over a period of 96 h to the lumpectomy bed plus a 2-cm margin. Perioperative complications, cosmetic outcome, and local control were assessed. RESULTS: The median follow-up for all patients is 20 months. Three patients experienced minimal perioperative pain that required temporary nonnarcotic analgesics. There have been four postoperative infections which resolved with oral antibiotics. No significant skin reactions related to the implant were noted and no patient experienced impaired would healing. Early cosmetic results reveal minimal changes consisting of transient hyperpigmentation of the skin at the puncture sites and temporary induration in the tumor bed. Good to excellent cosmetic results were noted in all 19 patients followed up a minimum of 24 months posttherapy. To date, 51 women have obtained 6-12-month follow-up mammograms and no recurrences have been noted. All patients currently have no physical signs of recurrence, and no patient has failed regionally or distantly. CONCLUSION: Treatment of the tumor bed alone with LDR interstitial brachytherapy appears to be well tolerated, and early results are promising. Long-term follow up of these patients is necessary to establish the equivalence of this treatment approach compared to standard BCT, however. PMID- 9226317 TI - Results of 3D conformal radiotherapy in the treatment of localized prostate cancer. AB - PURPOSE: 3D conformal radiotherapy (3D CRT) has been shown to decrease acute morbidity in the treatment of prostate cancer. Therapeutic outcome and late morbidity data have been accumulating. To evaluate the results of 3D CRT for the treatment of prostate cancer, we analyzed the outcome of a large series of patients treated with conformal techniques. MATERIAL AND METHODS: From January 1987 through June 1994, 707 patients with localized prostate cancer were treated with 3D CRT. Patients with pathologically-confirmed pelvic lymph node metastasis, treated with pre-irradiation (preRT) androgen ablation, or treated post prostatectomy were excluded. All had CT obtained specifically for treatment planning, multiple structures contoured on the axial images, and beam's-eye view conformal beams edited to provide 3D dose coverage. Median follow-up is 36 mos; 70 patients have been followed longer than 5.5 years. Six hundred three had T1-T2 tumors. PreRT prostate specific antigen (PSA) was available for 649 patients: median preRT PSA was 12.9 ng/ml, 209 patients had preRT PSA > 20 ng/ml. The median dose of radiation was 69 Gy; 102 patients received > or = 69 Gy. Biochemical failure was defined as: 1) two consecutive PSA rises over 2.0 ng/ml if nadir PSA < or = 2.0 ng/ml, 2) two consecutive PSA rises over nadir if nadir PSA > 2.0 ng/ml, or 3) initiation of hormonal therapy after RT. Complications were graded using the RTOG system. RESULTS: PreRT PSA and Gleason score emerged as independent indicators of biochemical control (bNED). Patients with preRT PSA > 10 had a significantly worse bNED at 5 years than patients with preRT PSA < or = 10. Five-year bNED was determined according to preRT PSA: PSA < or = 4, 88%; PSA > 4 < or = 10, 72%; PSA > 10 < or = 20, 43%; and PSA > 20, 30%. Patients with Gleason score > or = 7 also had a significantly worse bNED than patients with Gleason score < 7. Patients were divided into two prognostic groups: a favorable group with PSA < or = 10, Gleason score < 7, and T1-T2 tumors, and an unfavorable group with PSA > 10, Gleason score > or = 7 or T3-T4 tumors and studied for the effect of dose on bNED status. The bNED at 5 years was 75% for the favorable group and 37% for the unfavorable group. In addition, a group that might be considered a surgical subset was reviewed: patients with PSA < or = 10, Gleason score < or = 7, and T1-T2 tumors who were < 70 years old. This subset had an 84% 5-year bNED rate and 98% 5-year overall survival. Complications with the techniques used here are very low: 3% risk at 7 years of Grade 3-4 complications and 1% risk at 7 years of Grade 3 bladder complications (no Grade 4). CONCLUSION: 3D CRT allows for treatment of prostate cancers with a very low risk of complications. Patients with relatively early disease as defined by preRT PSA, Gleason score < 7, and T1-2 tumors and patients who are candidates for radical prostatectomy have excellent 5-year bNED rates. Patients with adverse prognostic factors have a high risk of biochemical recurrence and are candidates for innovative therapy. PMID- 9226318 TI - Initial clinical assessment of CT-MRI image fusion software in localization of the prostate for 3D conformal radiation therapy. AB - PURPOSE: To assess the utility of image fusion software and compare MRI prostate localization with CT localization in patients undergoing 3D conformal radiation therapy of prostate cancer. MATERIALS AND METHODS: After a phantom study was performed to ensure the accuracy of image fusion procedure, 22 prostate cancer patients had CT and MRI studies before the start of radiotherapy. Immobilization casts used during radiation treatment were also used for both imaging studies. After the clinical target volume (CTV) (prostate or prostate + seminal vesicles) was defined on CT, slices from the MRI study were reconstructed to precisely match the CT slices by identifying three common bony landmarks on each study. The CTV was separately defined on the matched MRI slices. Data related to the size and location of the prostate were compared between CT and MRI. The spatial relationship between the tip of urethrogram cone on CT and prostate apex seen on MRI was also estimated. RESULTS: The phantom study showed the registration discrepancies between CT and MRI smaller than 1.0 mm in any pair in comparison. The patient study showed a mean image registration error of 0.9 (+/- 0.6) mm. The average prostate volume was 63.0 (+/- 25.8) cm3 and 50.9 (+/- 22.9) cm3 determined by CT and MRI, respectively. The difference in prostate location with the two studies usually differed at the base and at the apex of the prostate. On the transverse MRI, the prostate apex was situated 7.1 (+/- 4.5) mm dorsal and 15.1 (+/- 4.0) mm cephalad to the tip of urethrogram cone. CONCLUSIONS: CT-MRI image fusion study made it possible to compare the two modalities directly. MRI localization of the prostate is more accurate than CT, and indicates the distance from cone to apex is 15 mm. CT-MRI image fusion technique provides valuable supplements to CT technology for more precise targeting of the prostate cancer. PMID- 9226319 TI - Prostate-specific antigen cancer volume: a significant prognostic factor in prostate cancer patients at intermediate risk of failing radiotherapy. AB - PURPOSE: Although the pretreatment serum prostate-specific antigen level (PSAL) is the single-most significant predictor of local and biochemical control in prostate cancer patients treated with radiotherapy, it is relatively insensitive for patients with a PSAL in the intermediate range (4-20 ng/ml). PSA density (PSAD) has been shown to be slightly more predictive of outcome than PSAL for this intermediate risk group; however, this improvement is small and of little use clinically. PSA cancer volume (PSACV), an estimate of cancer volume based on PSA, has recently been described and has been purported to be more significant than PSAL in predicting early biochemical failure after radiotherapy. We report a detailed comparison between this new prognostic factor, PSAL, and PSAD. METHODS AND MATERIALS: The records of 356 patients treated with definitive external beam radiotherapy for regionally localized (T1-4,Nx,M0) adenocarcinoma of the prostate were reviewed. Each patient had a PSAL, biopsy Gleason score, and pretreatment prostate volume by transrectal ultrasonography. The median PSAL was 9.3 ng/ml and 66% had Gleason scores in the 2-6 range. The median radiation dose was 66.0 Gy and the median follow-up for those living was 27 months. PSACV was calculated using a formula which takes into account PSAL, pretreatment prostate ultrasound volume, and Gleason score. The median PSACV was 1.43 cc. Biochemical failure was defined as increases in two consecutive follow-up PSA levels, one increase by a factor > 1.5, or an absolute increase of > 1 ng/ml. Local failure was defined as a cancer-positive prostate biopsy, obtained for evidence of tumor progression. RESULTS: The distributions of PSACV and PSAL were similar and, when normalized by log transformation, were highly correlated (p < 0.0001, linear regression). There was a statistically significant relationship between PSACV and several potential prognostic factors including PSAL, PSAD, stage, Gleason score, and pretreatment prostatic acid phosphatase (PAP). In univariate analyses, PSACV, PSAL, and PSAD proved to be the most significant predictors of both biochemical and local control. In multivariate analyses using Cox proportional hazards models with PSAL, PSAD, PSACV, and PAP as continuous variables, PSAL, PSACV, and Gleason score were significant in predicting biochemical control. Only PSAL was significantly correlated with local control. However, when these analyses were restricted to patients with intermediate PSALs (4-20 ng/ml), only PSACV was significant for predicting both biochemical and local control. CONCLUSION: PSACV was highly correlated with actuarial local and biochemical control and was superior to both PSAL and PSAD in predicting these outcomes in patients with PSALs between 4 and 20 ng/ml. PMID- 9226320 TI - Effect of edema on the post-implant dosimetry of an I-125 prostate implant: a case study. AB - PURPOSE: To investigate the effect of post-implant edema on the CT-based calculation of the total dose delivered by an I-125 prostate implant. MATERIALS AND METHODS: CT scans of a transperineal I-125 prostate implant were obtained 1 and 39-days post-implant. Changes in the prostate dimensions were determined from changes in the spatial distribution of the I-125 seeds. The total dose delivered to the target volume was computed from each CT scan, and the results compared. RESULTS: The volume of the prostate decreased by approximately 17% during the 38 day interval between the first and second CT scans. As a result, the radiation dose computed from the second CT scan was 13% higher. CONCLUSION: Post-implant edema can cause a significant underestimation of the radiation dose delivered by an I-125 prostate implant. Similar analysis should be carried out among a larger cohort of patients to confirm or refute these observations. PMID- 9226321 TI - A simple method to stabilize the prostate during transperineal prostate brachytherapy. AB - PURPOSE: The authors report a simple, remarkably effective method to limit prostatic motion during prostate brachytherapy. METHODS AND MATERIALS: Two 18-ga. stainless-steel needles (the same type used for source placement) are inserted transperineally, obliquely into the prostate, using finger and fluoroscopic guidance. These needles are left in place during the implant procedure. In our experience, maximum lateral displacement is decreased from about 1 cm to 0.2 mm, while craniocaudal motion is virtually eliminated. CONCLUSION: The method described here is simple to perform and does not require specialized equipment. PMID- 9226322 TI - Seminoma arising in corrected and uncorrected inguinal cryptorchidism: treatment and prognosis in 66 patients. AB - PURPOSE: The purpose of this study was to analyze prognosis and treatment results for seminoma arising in corrected and uncorrected inguinal cryptorchidism (SCIC and SUIC). METHODS AND MATERIALS: We reviewed 66 patients with inguinal seminomas between June 1958 and December 1991 at the Cancer Hospital and Institute of Chinese Academy of Medical Sciences. Of these patients, 23 had prior orchiopexy and 43 presented with an inguinal form of cryptorchidism. At presentation, 17 of 66 (26%) patients had nodal metastases. This nodal involvement was 30% (7 of 23) for SCIC and 23% (10 of 43) for SUIC, respectively. These numbers are comparable with those in a series of patients treated for scrotal seminoma at our institution (26% vs. 20%). However, 3 of 23 (13%) patients who had prior orchiopexy presented with inguinal nodal metastasis as compared with 0 of 43 patients with SUIC or 4 of 237 patients with scrotal seminoma (p < .05). There were 49 stage I, 5 stage IIA, 8 stage IIB, 3 stage III, and 1 stage IV patients. All patients underwent radical orchiectomy and received further radiotherapy, chemotherapy, or both. Patients with stage I and stage II disease were treated primarily with radiotherapy, whereas patients with stage III and IV disease were treated with chemotherapy. RESULTS: The overall and disease-free survival at 5 and 10 years was 94% and 92%, 89% and 87%, respectively. The overall 5- and 10 year survival by stage was 100% and 100% for stage I, and 77% and 68% for stage II, respectively (p < .05). There was no significant difference in survival between SUIC and SCIC (93% vs. 96% at 5 years). Four patients developed relapse. Two of these four patients experienced relapse at the inguinal area, due to a marginal miss. Three of four patients with relapse were successfully salvaged, and one died of disease. CONCLUSION: Our results indicate that prognosis for inguinal seminoma is excellent and similar to that of scrotal seminoma. Postorchiectomy radiotherapy can be considered as the standard treatment for stage I and IIA inguinal seminoma. We recommend routinely including the para aortic and ipsilateral pelvic nodes. PMID- 9226323 TI - Clinical characteristics, prognosis, and treatment of pelvic cryptorchid seminoma. AB - PURPOSE: To analyze the clinical characteristics, prognosis, and treatment outcome of pelvic cryptorchid seminoma (PCS), and to determine whether whole abdominal-pelvic irradiation for Stage I disease is necessary. METHODS AND MATERIALS: From 1958 to 1991, 60 patients with PCS were treated at the Cancer Hospital of Chinese Academy of Medical Sciences, Beijing. They presented with a lower abdominal mass and showed a predominance for the right side. A high proportion of patients with PCS [26 of 60 (43%)] had metastatic disease, compared to 20% of those with scrotal seminoma, and there was a tendency toward a higher frequency of pelvic nodal metastases. There were 34 Stage I, 6 Stage IIA, 11 Stage IIB, 5 Stage III, and 4 Stage IV patients. Of these 60 patients, 56 underwent laparotomy with or without cryptorchiectomy (37 radical orchiectomy, 7 partial orchiectomy, and 12 biopsy of the primary or cervical node), and 4 cervical node biopsy only. All patients were further treated with radiotherapy, chemotherapy, or a combination of both. Patients with Stage I and II disease received radiotherapy, whereas patients with Stage III and IV were treated with chemotherapy. RESULTS: The overall and disease-free survivals at 5 and 10 years were 92% and 87%, and 88% and 84%, respectively. The 5- and 10-year survivals were 100% for Stage I, 94% and 87% for Stage II, and 56% and 42% for Stage III/IV, respectively (p < 0.05). Volume of irradiation, i.e., whole abdominal pelvic radiotherapy (10 patients), versus hockey-stick encompassing paraaortic, ipsilateral iliac nodes and the primary tumor or tumor bed (17) did not influence outcome in Stage I patients. Five patients relapsed within 2-12 years after treatment, and four of these patients were successfully salvaged. Four patients developed a second malignant tumor and died. CONCLUSION: Stage I and II PCS can be adequately controlled by radiotherapy regardless of the surgical procedure. Whole abdominal-pelvic irradiation for Stage I and IIA disease is not required, and fields can be limited to the paraaortic, ipsilateral iliac nodes and primary tumor or tumor bed. We recommend platinum-based chemotherapy for Stage IIB-IV PCS. PMID- 9226324 TI - Clinical results of radiofrequency hyperthermia for malignant liver tumors. AB - PURPOSE: To evaluate thermometry and the clinical results of radiofrequency (RF) hyperthermia for advanced malignant liver tumors. METHODS AND MATERIALS: One hundred seventy-three patients with malignant liver tumors treated between 1983 and 1995 underwent hyperthermia. The 173 tumors consisted of 114 hepatocellular carcinomas (HCCs) and 59 non-HCCs (47 metastatic liver tumors and 12 cholangiocarcinomas). Eight-megahertz RF capacitive heating equipment was used for the hyperthermia. Two opposing 25-cm electrodes were generally used for heating the liver tumors. Our standard protocol was to administer hyperthermia 40 50 min twice a week for a total of eight sessions. The liver tumor temperature was measured by microthermocouples when possible. Transcatheter arterial embolization, radiotherapy, immunotherapy, and chemotherapy were combined with hyperthermia treatment in accordance with each patient's liver function. RESULTS: One hundred forty (81%) of the 173 patients who underwent more than four sessions of hyperthermia were evaluated in this study. Thermometry was performed in 77 (55%) of these 140 patients. The maximum tumor temperature, average tumor temperature, and minimum tumor temperature in the HCC were (mean +/- standard error) 41.2 +/- 0.2 degrees C, 40.3 +/- 1.3 degrees C, and 40.1 +/- 0.2 degrees C, respectively. The same thermometry results for non-HCC were 42.3 +/- 0.2 degrees C, 41.2 +/- 0.2 degrees C, and 40.9 +/- 0.2 degrees C, respectively. The maximum and minimum temperatures (41.8 +/- 0.2 degrees C and 40.3 +/- 0.4 degrees C) in the patients with a complete or partial response (CR or PR) were higher than those in the patients with no response or progressive disease (NR or PD) (41.3 +/- 0.5 degrees C and 39.8 +/- 0.4 degrees C), but the difference was not significant. Of the 73 cases with HCC who were evaluated by computed tomography (CT), CR was achieved in 7 (10%), PR in 15 (21%), NR in 37 (51%), and PD in 14 (19%). Of the 45 cases involving liver metastases evaluated by CT, CR was achieved in 3 (7%), PR in 17 (38%), NR in 12 (27%), and PD in 13 (29%). The 1 year cumulative survival rate for HCC patients was 30.0%, and the 5-year survival rate was 17.5%. The 1-year survival of non-HCC patients was 32.5%, and the longest survival was 30 months. The sequelae of hyperthermia included focal fat necrosis in 20 patients (12%), gastric ulceration in 4 (2%), and liver necrosis in 1 (1%). The sequelae of thermometry were severe peritoneal pain in seven patients (11%), intraperitoneal hematoma in one (1%), and pneumothorax in one (1%). CONCLUSION: Even though the thermometry results for liver tumors were not satisfactory, the treatment results are promising. Further clinical trials of RF capacitive hyperthermia for the treatment of advanced liver tumors should be encouraged. PMID- 9226325 TI - Radiation tolerance of cirrhotic livers in relation to the preserved functional capacity: analysis of patients with hepatocellular carcinoma treated by focused proton beam radiotherapy. AB - PURPOSE: To determine the preserved functional capacity of the liver as a probable determinant of radiation tolerance in patients with cirrhosis and hepatocellular carcinoma, who underwent proton beam radiotherapy. MATERIALS AND METHODS: We reviewed computed tomographic (CT) scans of 26 patients with cirrhosis and hepatocellular carcinoma during a period of 12-27 months after proton beam radiotherapy. Tumors were treated with focused proton beams with target doses of 140-186 TDF (time, dose, and fractionation). We measured the degree of hypertrophy of the untreated liver volume by measuring the total liver volume and the treated liver volume which was radiologically identified on contrast-enhanced CT scans. The risk of radiation-induced liver failure was estimated using the prediction score (PS) originally used for estimating posthepatectomy liver failure, substituting the planned treated liver volume for the resection liver volume. RESULTS: The degree of hypertrophy ranged from -19% to 51%, and was significantly positively correlated with the ratio of the planned treated liver volume to the functional liver volume. The PS agreed well with observed radiation tolerance in 21 patients, but underestimated the tolerance in 5. This underestimation was diminished when the PS was recalculated using the identified untreated liver volume in lieu of the planned untreated liver volume. CONCLUSION: As in surgical treatment, radiation tolerance of the cirrhotic liver after focused treatment is closely related to the preserved functional capacity of the identified untreated liver volume, which shows compensatory hypertrophy following radiotherapy. PMID- 9226327 TI - Adjuvant radiation for vulvar carcinoma: improved local control. AB - PURPOSE: Local recurrence is a significant problem following primary surgery for advanced vulva carcinoma. The objectives of this study were to evaluate the impact of adjuvant vulvar radiation on local control in high risk patients and the impact of local recurrence on overall survival. METHODS AND MATERIALS: From 1980-1994, 62 patients with invasive vulva carcinoma and either positive or close (less 8 mm) margins of excision were retrospectively studied. Thirty-one patients were treated with adjuvant radiation therapy to the vulva and 31 patients were observed after surgery. Kaplan-Meier estimates and the Cox proportional hazard regression model were used to evaluate the effect of adjuvant radiation therapy on local recurrence and overall survival. Independent prognostic factors for local recurrence and survival were also assessed. RESULTS: Local recurrence occurred in 58% of observed patients and 16% in patients treated with adjuvant radiation therapy. Adjuvant radiation therapy significantly reduced local recurrence rates in both the close margin and positive margin groups (p = 0.036, p = 0.0048). On both univariate and multivariate analysis adjuvant radiation and margins of excision were significant prognostic predictors for local control. Significant determinants of actuarial survival included International Federation of Gynecologists and Obstetricians (FIGO) stage, percentage of pathologically positive inguinal nodes and margins of excision. The positive margin observed group had a significantly poorer actuarial 5 year survival than the other groups (p = 0.0016) and adjuvant radiation significantly improved survival for this group. The 2 year actuarial survival after developing local recurrence was 25%. Local recurrence was a significant predictor for death from vulva carcinoma (risk ratio 3.54). CONCLUSION: Local recurrence is a common occurrence in high risk patients. In this study adjuvant radiation therapy significantly reduced local recurrence rates and may improve overall survival in certain subgroups. As salvage rates after developing local recurrence are poor adjuvant vulvar radiation should be considered for patients at risk after primary surgery. PMID- 9226326 TI - Excellent long-term survival and absence of vaginal recurrences in 332 patients with low-risk stage I endometrial adenocarcinoma treated with hysterectomy and vaginal brachytherapy without formal staging lymph node sampling: report of a prospective trial. AB - PURPOSE: The value of adjuvant radiation therapy and staging pelvic lymphadenectomy in patients with low-risk, early-stage endometrial cancer is controversial. The aim of this study was to report the long-term survival, rate of recurrences, and complications in patients with Stage I endometrial cancer, Grade 1-2, with <50% myometrial invasion treated with hysterectomy (without formal staging pelvic and periaortic lymph node sampling or lymphadenectomy) and postoperative vaginal brachytherapy. METHODS AND MATERIALS: A total of 303 patients with pathologic Stage I endometrial cancer, Grade 1-2, with <50% myometrial invasion and nonmalignant peritoneal cytology, were treated with total abdominal hysterectomy, bilateral salpingo-oophorectomy, and postoperative vaginal brachytherapy (30 Gy to point 0.5 cm depth) in a prospective study extending from 1958 to 1994. In addition, 29 additional Stage I, Grade 1-2 patients with <50% myometrial invasion and malignant peritoneal cytology were treated with 1 year of progesterone therapy. Patients were followed for 1.2-32 years (median 8.1 y). RESULTS: Six patients had recurrences and died secondary to disease. There were no vaginal recurrences. The 5-, 10-, 20-, and 30-year disease free survivals of the 303 patients with nonmalignant peritoneal cytology were 98.9%, 97.8%, 96.7%, and 96.7%, respectively. Patients with malignant peritoneal cytology had a 5- and 10-year disease-free survival of 100%. Significant radiation complications occurred in 2.1% of the patients. CONCLUSION: In patients with low-risk, Stage I endometrial cancer, hysterectomy and adjuvant postoperative vaginal brachytherapy provide excellent long-term survival, eliminate vaginal recurrences, and are not associated with significant complications. The addition of 1 year of progesterone therapy to patients with malignant cytology provides 100% long-term survival. Based on these results, patients with low-risk, Stage I endometrial adenocarcinoma do not need formal staging pelvic and periaortic lymphadenectomy. PMID- 9226328 TI - High-dose-rate intracavitary brachytherapy (HDR-IC) in treatment of cervical carcinoma: 5-year results and implication of increased low-grade rectal complication on initiation of an HDR-IC fractionation scheme. AB - PURPOSE: To report the treatment results and rectal/bladder complications of cervical carcinoma radically treated with high-dose-rate intracavitary brachytherapy (HDR-IC). The current policy of using three-fraction scheme was examined. METHODS AND MATERIALS: Between November 1987 and August 1990, 173 patients with cervical carcinoma were treated with curative-intent radiation therapy. Whole pelvic irradiation was administered with 10-MV X ray. Dose to the central cervix was 40-44 Gy in 20-22 fractions, following by pelvic wall boost 6 14 Gy in three to seven fractions with central shielding. 60Co sources were used for HDR-IC, and 7.2 Gy was given to Point A for three applications, 1-2 weeks apart. Duration of follow-up was 5-7.8 years. RESULTS: Twenty-eight patients (16%) developed central-regional recurrences. Overall 5-year actuarial pelvic control rate was 83%. By stage, 5-year actuarial pelvic control rates were 94%, 87%, and 72% for Stages IB + IIA, IIB + IIIA, and IIIB + IVA, respectively. Thirty-one patients (18%) developed distant metastasis. Overall 5-year actuarial survival rate was 58%. By stage, 5-year actuarial survival rates were 79%, 59%, and 41% for Stages IB + IIA, IIB + IIIA, and IIIB + IVA, respectively. Sixty-six (38%) and 19 patients (11%) developed rectal and bladder complications, respectively. For rectal complication, the overall actuarial rate was 38% at 5 years. By grade, 5-year actuarial rectal complication rates were 24%, 15%, 4%, and 3% for Grades 1-4, respectively. Overall prevalence of rectal complications was 37% and 14% at 2 and 5 years, respectively. Prevalence of low-grade rectal complication (Grades 1 and 2) was dominant at 2 years (30%), but declined to 8% at 5 years. Prevalence of high-grade, severe rectal complication (Grades 3 and 4) remained steady at 2 and 5 years (7% and 6%, respectively). Five-year actuarial bladder complication was 9%. Five-year prevalence of bladder complication was 2%. CONCLUSION: Using a three-fraction scheme, survival rate appeared comparable with the existing results of the low-dose-rate technique. The incidence of rectal complication with this scheme remained relatively high. The increased part of rectal complication was predominantly low grade. This result suggested that therapeutic gain with this scheme may not be good enough to circumvent its biologic disadvantage. Numbers of fractions >3 must be considered in future trials. PMID- 9226329 TI - Quantification of radiation-induced regional lung injury with perfusion imaging. AB - PURPOSE: To better understand the dose and time dependence of radiation therapy (RT)-induced regional lung dysfunction as assessed by changes in regional lung perfusion. METHODS AND MATERIALS: Patients who were to receive RT for tumors in and around the thorax, wherein portions of healthy lung would be incidentally irradiated, were prospectively studied. Regional function was assessed pre- and post-RT with single photon emission computed tomography (SPECT) lung perfusion scans, obtained following the intravenous administration of approximately 4 mCi of technetium-99m macroaggregated albumin. Pre-RT computed tomography (CT) scans were used to calculate the three-dimensional (3D) dose distribution, reflecting tissue density inhomogeneity corrections. Each SPECT scan was correlated with the pre-RT CT scan, and the 3D dose distribution. Changes in regional lung perfusion were correlated with regional RT dose, at various time intervals following radiation. RESULTS: The data from 20 patients (7 breast cancer, 5 lymphoma, 1 esophagus, 1 sarcoma, and 6 lung cancer) have been analyzed. Patients with gross intrathoracic lung cancers causing obstruction of regional pulmonary arteries were not included. For most patients, there is a statistically significant dose dependent reduction in regional blood flow at all time points following radiation. While a time dependence is suggested in the high dose range, the limited amount of data prevents meaningful statistical evaluation. CONCLUSIONS: Radiation therapy-induced regional lung dysfunction occurs in a dose-dependent manner and develops within 3-6 months following radiation. In contrast to classical "sigmoid" dose-response curves, described mainly for changes following whole lung irradiation, these data suggest a more gradual relationship between regional dysfunction and RT dose. Retraction of irradiated lung with secondary movement of unirradiated lung into the "3D-defined irradiated volume" may have introduced inaccuracies into this analysis. Additional studies are currently underway to assess this possibility and better refine this dose-response curve. Studies are underway to determine if changes in assessments of whole lung function, such as pulmonary function tests, can be predicted by summing the regional changes observed. PMID- 9226330 TI - Induction thermochemotherapy increases therapeutic gain factor for the fractionated radiotherapy given to a mouse fibrosarcoma. AB - PURPOSE: It has been shown that thermochemotherapy (TC) given prior to radiation reduces the number of clonogens, with a resultant decrease in the tumor control radiation dose. The purpose of this article was to investigate using an animal tumor model how this clonogen reduction affects subsequent fractionated radiotherapy, including repopulation of surviving clonogens, and whether the induction TC can increase the therapeutic gain factor (TGF). METHODS AND MATERIALS: The single-cell suspensions prepared from the fourth-generation isotransplants of a spontaneous fibrosarcoma, FSa-II, were transplanted into the C3Hf/Sed mouse foot. TC was given by heating tumors at 41.5 degrees C for 30 min immediately after an intraperitoneal injection of cyclophosphamide (200 mg/kg) when tumors reached an average diameter of 4 mm. Fractionated radiotherapy (R) with equally graded daily doses was initiated 24 h after TC either in air (A) or under hypoxic conditions (H). The 50% tumor control dose (TCD50) and the radiation dose to induce a score 2.0 reaction (complete epilation with fibrosis) in one-half of irradiated animals, RD50(2.0), were obtained, and the TGF was calculated. Our previous results on the fractionated radiotherapy using the same tumor system served as controls. RESULTS: The TCD50(A, single dose) and TCD50(H, single dose) following TC+R were 52.2 and 57.3 Gy, respectively, which were 14.0 and 20.4 Gy lower than those following radiation alone. The TCD50(A, TC+R) increased only slightly when the number of fractions was increased from one to 10 doses, and all TCD50s were significantly lower than the TCD50(A, R alone). Both TCD50(H, TC+R) and TCD50(H, R alone) increased consistently from a single dose to 20 doses, but all TCD50(H, TC+R) were significantly lower than the TCD50(H, R alone). Regarding the normal tissue reaction, the RD50 values both following TC+R and R alone increased consistently from a single dose to 20 daily doses. However, the RD50(TC+R) and RD50(R alone) for each corresponding number of fractions was not significantly different, resulting in the TGFs significantly > 1.0 for combined TC+R treatments, with the exception of 20 daily doses given in air. CONCLUSION: The induction TC decreased the TCD50 values substantially without altering the RD50 for a late reaction, resulting in an significant increase in the TGF. These results encourage the use of TC as an induction treatment prior to fractionated radiotherapy. PMID- 9226331 TI - Addition of cisplatin improves efficacy of 131I-labeled monoclonal antibody 323/A3 in experimental human ovarian cancer. AB - This study was conducted to determine whether the cytotoxic agent cisplatin (CDDP), also known as a radiosensitizer, can improve the efficacy of the 131I labeled monoclonal antibody (MAb) 323/A3 in the treatment of experimental human ovarian cancer. METHODS AND MATERIALS: Nude mice bearing well-established subcutaneous FMa, OVCAR-3, or Ov.Pe xenografts were injected twice with a 2-week interval either with a bolus of CDDP, 131I-323/A3, or with a combination of both modalities. CDDP was injected at various timepoints when combined with 131I 323/A3. The efficacy of the treatment was expressed as the specific growth delay (SGD). The growth inhibitory effect of the combination was characterized to detect additivity or synergism, using the mean relative tumor volumes at 2, 4, and 6 weeks after the last injection as endpoints. RESULTS: The efficacy of 131I 323/A3 was superior to that of the maximum tolerated dose (MTD) of CDDP (6 mg/kg) in all three xenografts. The addition of CDDP to 131I-323/A3 could increase the growth inhibition in the CDDP-responsive FMa and OVCAR-3 xenografts, but not in Ov.Pe xenografts. Although this improved antitumor effect was additive rather than synergistic, the combination was more effective when compared with that of the MTD of each of the modalities alone. The time interval between the administration of a bolus injection of CDDP and 131I-323/A3 had no effect on the extent of growth inhibition in OVCAR-3 xenografts. CONCLUSION: The addition of CDDP to 131I-323/A3 resulted in an additive inhibitory effect on the growth of CDDP-responsive xenografts. As the combination of radioimmunotherapy and CDDP was more effective in the inhibition of the tumor growth when compared with that of the MTD of each of the modalities alone, this treatment may therefore be considered of use in patients with ovarian cancer responsive to CDDP. PMID- 9226332 TI - Rapid assay of intrinsic radiosensitivity based on apoptosis in human CD4 and CD8 T-lymphocytes. AB - PURPOSE: An assay for radiosensitivity has numerous applications in the clinic. Avoidance of acute responses, prediction of normal tissue toxicity, and individualization of patient radiotherapy are included among these. We have developed a rapid assay (about 24 h) able to predict intrinsic radiosensitivity of CD4 and CD8 T-lymphocytes based on radiation-induced apoptosis. METHODS AND MATERIALS: Fresh blood samples (1-2 ml in heparinized tubes) were irradiated with 0-, 2-, and 8-Gy X rays at a dose rate of approximately 3 Gy/min. Following irradiation, the cells were collected and prepared for flow-cytometric analysis and cell sorting. In conjunction with the CellQuest software available with the FACSVantage cell sorter (Becton-Dickinson), two T-lymphocyte types were analyzed on the basis of their cell-specific antigens (CD4 and CD8), and DNA was stained with DAPI. Following the separation of these cell types, radiation-induced cell death was assessed. Cytotoxicity was characterized by gradual degradation of internucleosomal DNA which results in a sub-G1 peak on the DNA histogram, and by the associated loss of surface antigens causing an intermediate positive peak in the antibody histogram. Using the assay, we investigated the interdonor variation in a cohort of 45 healthy adult blood donors and 5 children [one had immunodeficiency, centromeric instability, and facial anomalies syndrome (ICF), and one had ataxia telangiectasia (AT)]. Intradonor variation was assessed with 10 different experiments from a single donor. RESULTS: CD4 and CD8 T-lymphocyte radiosensitivities were correlated (r = 0.63 and 0.65 for 2 and 8 Gy, respectively) in 45 adult donors. Both for CD4 and CD8 cells, 2 and 8 Gy irradiation responses showed a good correlation (r = 0.77 for both). Interdonor variation was significantly higher than intradonor variation (p < 0.0005) for all CD4 and CD8 data. We observed a decrease in the antigen fluorescence of dying cells, a phenomenon referred to as antigen-ebb. Antigen-ebb was clearly observed in both cell types, and correlated significantly with cytotoxicity. A trend was observed between radiosensitivity and donor age, but there was no correlation for gender. Blood from a 4-year-old girl presenting with ICF demonstrated compromised radiation-induced cytotoxicity in her CD4 T-lymphocytes, and an 11-year-old boy presenting with AT demonstrated compromised radiation-induced cytotoxicity in both his CD4 and CD8 T-lymphocytes. CONCLUSION: We conclude that the assay provides a rapid means of determining radiosensitivity, can discriminate differences in radiation-induced cytotoxicity between individuals, and can be used as a rapid screen for genetically hypersensitive patients. Antigen-ebb offers interesting possibilities for molecular biological investigations, permitting characterization and isolation of abnormal but vital cells in the absence of clastogenic agents. PMID- 9226333 TI - 1995 survey of physics teaching efforts in radiation oncology residency programs. AB - A physics teaching survey was constructed and sent to the 83 radiation oncologist training programs. The survey requested program information regarding size, staffing, curriculum, lab/rotation programs, organization, requirements, instructor makeup, teaching materials, and board certification examination results. The surveys were sent to the physicist responsible for the physics program. Forty-nine (59%) institutions returned completed surveys, of which 43 (88%) were university-associated programs, and 27 (55%) were 4-year programs. On average, there were two residents/year. Most programs (39) taught physics exclusively during the first year (PG2). Some programs taught different subjects (or levels) to different year residents. Radiation dosimetry, treatment planning, and brachytherapy constituted nearly half of the teaching hours. On average the total classroom time expended by physicists was 61.4 h/year with a range of 24 118 h. The mean for laboratory/demonstration time was 27 h/year with 18 programs providing none. Physics orientation/rotations ranged from 1 to 480 h with a mean of 170 h for a physics rotation taking place in year 2 (PG3). Mandatory attendance was 80% for first-year residents and decreased in later years. Homework was assigned in 76% of the programs, and 65% of the programs were graded. The primary instructors averaged 18.2 years of experience, and the majority were ABR/ABMP certified. Khan's textbook was the most prevalent resource for most subjects. No correlation could be made between teaching hours and ABR physics percentile scoring. The survey results reveal enormous differences in national teaching efforts. PMID- 9226334 TI - Measurement of 6-MV X-ray surface dose when topical agents are applied prior to external beam irradiation. AB - PURPOSE: Radiation therapy patients are typically warned not to apply lotions, deodorants, or powders to the skin within the treatment area because of the possible increase in surface dose due primarily to a bolus effect. This study investigates the effect of 15 products, with and without high atomic number components, on surface dose. METHODS AND MATERIALS: A Markus-type parallel plate ionization chamber in a polystyrene phantom was used to measure surface doses for normal applications of the products for a small (5 x 5-cm2) and a large (25 x 25 cm2) field size. RESULTS: The greatest surface dose increase for any product was 5.4% (21.8-27.2%) of the d(max) dose for the small field and 1.0% (43.6-44.6%) for the large field. Products with high-atomic-number components did not increase the surface dose relative to radiation therapy specialty products. CONCLUSION: No large increase in surface dose was detected with a normal application of the products. However, the possibility exists that an increase in skin reaction may occur owing to chemical irritants in the applied product. PMID- 9226335 TI - Beta-endorphin and cortisol levels in plasma and CSF following acute experimental spinal traumas. AB - beta-endorphin and cortisol were measured in cerebrospinal fluid (CSF) and plasma by radioimmunological method (RIA) in two groups of rabbits with spinal cord traumatic injuries at cervical and lumbar levels, respectively with and without concomitant spinal shock and arterial hypotension, and in a group of sham operated animals as controls. The two groups with spinal lesions displayed a significant beta-endorphin increase in CSF, whereas the cortisol level remained unchanged both in the spinal traumatized rabbits and in controls. Both the opioid and the cortisol concentration rose significantly in plasma in all three groups and in particular resulted significantly higher in the cervical traumatized group where spinal trauma was associated with spinal shock and hypotension. However, no significant difference was found when beta-endorphin concentrations in plasma were compared between the sham operated animals and the spinal lumbar traumatized animals without concomitant spinal shock. The results seem to suggest that the beta-endorphin increase in CSF is related to the nervous tissue lesion, while its increase in plasma, like that of cortisol, is due to surgery or other stress factors inherent in the experiment. This independent behaviour of beta-endorphin in plasma and in CSF suggests its different origin in these two compartments. PMID- 9226336 TI - Is there a role for corticosterone in expression of abnormal behaviour in restricted-fed fowls? AB - Growing parent stock (breeders) of meat-type chickens (broilers), subjected routinely to chronic food restriction, show increased pacing before a single daily meal and increased drinking and pecking at non-food objects (oral stereotypies) afterwards. Expression of these activities is correlated positively with the level of restriction imposed, and is thought to be controlled mainly by central dopaminergic mechanisms. There is published evidence that glucocorticoids can amplify dopamine mediated behaviours, and this paper describes four experiments examining the relationship between corticosterone and behaviour in individually caged broiler breeders. In Experiment 1 (with 3 levels of food restriction), plasma corticosterone concentration was correlated positively with the level of restriction imposed when blood samples were taken in the morning, but not when they were taken in the afternoon. This may be because corticosterone level declines from morning to afternoon with mild but not severe restriction. In Experiment 2 (severe restriction only), plasma corticosterone level did not change significantly with time of day, and mean values of individual birds were not correlated with their observed times spent in oral stereotypies. In Experiment 3 (severe restriction), object pecking increased in a dose-related way after systemic injection of 1-4 mg/kg corticosterone (significant) and 7.5-30 IU/kg ACTH (not significant), and 10-40 mg/kg metyrapone (corticosterone synthesis inhibitor) had no effect. In Experiment 4 (severe restriction), 180 mg/day metyrapone administered in food for 5 days reduced the plasma corticosterone response to injection of 15 IU/kg ACTH on the fourth day, but otherwise had no effect. Significant increases in drinker directed activity after injection of ACTH on the fourth day and 4 mg/kg corticosterone on the fifth day coincided with greatly elevated plasma levels of corticosterone. It is concluded that the oral stereotypies of restricted-fed broiler breeders do respond to induced increases in plasma corticosterone concentration that are supra physiological, but there may be only a weak association between behaviour and corticosterone within the physiological range. PMID- 9226337 TI - Effects of naltrexone on food intake and changes in subjective appetite during eating: evidence for opioid involvement in the appetizer effect. AB - The effects of 50 mg naltrexone on eating and subjective appetite were assessed in a double-blind placebo-controlled study with 20 male volunteers. Appetite was monitored using a disguised digital balance connected to a micro-computer, which constantly monitored the amount of food remaining, and which automatically interrupted feeding for 30 s after every 50 g consumed to allow appetite ratings to be made. Half the subjects ate pasta with a cheese sauce, and the remainder pasta with a tomato sauce. Subjects ate significantly less of both foods after 50 mg naltrexone than in either the placebo condition or on the initial (familiarisation) day. Naltrexone also reduced the rated pleasantness of both foods, and reduced overall eating rate. When best-fit quadratic functions were used to describe changes in rated hunger in relation to intake within the meal, naltrexone abolished the positive linear component reflecting the initial stimulation of appetite without altering either intercept or the negative quadratic function. Although mood ratings suggested that naltrexone had a mild sedative effect, mood changes alone could not explain the effects of naltrexone on appetite. Overall, these data suggest a specific role for opioids in the stimulation of appetite through palatability. PMID- 9226338 TI - The effect of anabolic-androgenic steroids on sexual behavior and reproductive tissues in male rats. AB - This study assessed the effects of high doses of anabolic-androgenic steroids (AAS) and their withdrawal on male reproductive behavior and reproductive tissues during development. Prepubertal, peripubertal, and adult male Long Evans rats were divided into 4 groups: 1) Testosterone propionate for 16 weeks (TP), 2) TP for 3 weeks and withdrawn for 13 weeks (TPWL), 3) TP for 16 weeks and withdrawn for 3 weeks (TPWS), 4) propylene glycol (control vehicle) for 16 weeks (PG). As determined by sexual performance and sexual preference tests, administration of high doses of AAS to the peripubertal animals enhanced sexual performance and sexual motivation. There was no significant effect on sexual behavior of the prepubertal animals. High doses of anabolic-androgenic steroids depleted Leydig cell number in the prepubertal and adult rats, but had no effect on the Leydig cell number of the peripubertal animals. After long-term withdrawal from AAS no significant effects on sexual behavior were found. The depletion of Leydig cells that occurred in the prepubertal animals after withdrawal was reversible, while the depletion of the Leydig cells of the adult animals did not return to the control level suggesting a long lasting alteration. PMID- 9226339 TI - Endogenous mu opioid systems and perioral responsiveness in the rat fetus. AB - Pharmacological manipulation of mu opioid receptors located in rostral and caudal parts of the brain produces distinctive changes in perioral responsiveness to nipple-like tactile stimulation in the E20 rat fetus. Blockade of caudal mu opioid receptors by intracistema magna (I.C.) injection of the selective mu antagonist drug CTOP reduces appetitive responses directed toward the artificial nipple. In contrast, blockade of mu opioid receptors in the rostral part of the brain by intracerebroventricular (I.C.V.) administration of CTOP increases fetal responsiveness to perioral cutaneous stimulation including oral capture and grasping of the artificial nipple. This pattern of the results suggests that there are at least two functionally different neuronal populations of mu opioid receptor-containing neurons that are involved in the regulation of the perioral responsiveness in the E20 rat fetus. The caudal part of this mu opioid system increases perioral responsiveness while the rostral part of the system decreases responsiveness to nipple-like perioral stimulation. These findings suggest the possibility that mu opioid systems may play a functional role in regulating neonatal behavior at the nipple. PMID- 9226340 TI - The effect of caloric load and nutrient composition on induction of small intestinal satiety in dogs. AB - The influence of caloric load and nutrient composition on small intestinal satiety was investigated in six dogs with chronic esophageal fistulas. Dogs received small bowel infusion of a mixed nutrient liquid meal, a fat, carbohydrate, and a protein solution over two hours. Each infusion was given over a range of caloric densities which represented between 0 to 20% of the total 24 h caloric requirement for each animal. Satiety was measured by sham feeding at the end of the infusion. Infusion of 0.25 cal/ml (equivalent to 5% of the 24 h caloric requirement) of a liquid mixed nutrient meal (Isocal) significantly suppressed sham feeding (volume sham fed: control 265 +/- 28 ml/min; Isocal 0.25 cal/ml 218 +/- 52 ml/min, p < 0.05). Oleate and dextrose polymer also significantly reduced sham feeding at a caloric concentration of 0.25 cal/ml (volume sham fed: control 265 +/- 28 ml/min; oleate 112 +/- 28 ml/min; oleate 112 +/- 9 ml/min; dextrose polymer 190 +/- 11 ml/min), whereas peptone did not significantly suppress sham feeding until a solution with 0.5 cal/ml was given. These studies demonstrate that caloric load of intestinal nutrients must reach a threshold to produce satiety in sham feeding dogs. The threshold caloric load is different for the three nutrient groups, with fats requiring the lowest caloric load to produce satiety. These provide insight into the mechanisms by which small intestinal signals might contribute to regulation of the inter-meal interval. PMID- 9226341 TI - Na depletion-induced Na appetite of sheep is independent of brain angiotensin II. AB - Previous experiments indicated that the Na appetite of Na-deplete sheep is decreased by systemically administered captopril. The assumption that captopril does not readily cross the blood-brain barrier, lead to the conclusion that circulating ANG II acting in brain areas without a blood-brain barrier, i.e., circumventricular organs such as the subfornical organ or organum vasculosum of the lamina terminalis, contributes to Na appetite induced by Na depletion. The present experiments investigated the possibility that systemically administered captopril does, in fact, cross the blood-brain-barrier and thereby influence brain angiotensin II formation and that brain angiotensin II contributes to Na depletion-induced Na appetite of sheep. The results showed that systemically administered captopril blocked water intake caused by intracerebroventricular infusion of angiotensin I, and that Na depletion induced Na appetite was not decreased by intracerebroventricular infusion of various antagonists of the renin angiotensin system. Thus, the results suggest that although captopril crosses the blood-brain-barrier and can influence the formation of brain angiotensin II, brain angiotensin II is not involved in the Na appetite of Na-deplete sheep. PMID- 9226342 TI - Involvement of the cholinergic system and periaqueductal gray matter in the modulation of tonic immobility in the guinea pig. AB - Unilateral microinjection of carbachol (CCh, 1.0 microg/0.2 microl) into the ventrolateral periaqueductal gray matter (vPAG) increased the duration of tonic immobility (TI) episodes induced by postural inversion and by movement restriction maneuvers in adult male guinea pig (Cavia porcellus), while stimulation with the same drug at the same concentration into the dorsolateral and dorsomedial periaqueductal gray matter (dl/dmPAG) decreased the duration of TI. Pretreatment with atropine (7.6 microg/0.4 microl) showed that the action of CCh is mediated by muscarinic receptors in the ventrolateral PAG but not in the dorsomedial and dorsolateral regions. These data suggest that the PAG and the cholinergic system are involved in the modulation of TI episodes and that different regions of the guinea pig PAG play distinct roles in the organization of this behavior. PMID- 9226343 TI - Effects of progesterone on the sexual behavior of castrated, testosterone-treated male cynomolgus monkeys (Macaca fascicularis). AB - In male cynomolgus monkeys the synthetic progestin, medroxyprogesterone acetate (MPA), decreases testosterone (T) levels and sexual behavior, binds to progestin receptors in brain, and reduces by about 70% the uptake of [3H]androgens by both brain and genital tract tissues. To examine the behavioral effects of progesterone (P) itself, eight castrated, T-treated males were each tested twice weekly with an estrogenized female before, during, and after they were treated with two SC Silastic P implants. Data from six 4-week treatment periods were analyzed to facilitate comparisons with our previous data using MPA: i) baseline, ii) weeks 4-7 of P treatment, iii) weeks 8-11 of P treatment, iv) weeks 1-4 after P implants were removed, v) weeks 5-8 after P withdrawal, and finally vi) weeks 9 12 after P withdrawal (384 1 h behavior tests). Weekly blood samples (N = 192) were analyzed by radioimmunoassay to determine plasma levels of both T and P. P treatment, which resulted in high plasma P levels (about 44 ng/ml), produced decrements in measures of male sexual behavior and motivation that were both qualitatively and quantitatively similar to those produced by MPA treatment but, unlike MPA, P did not decrease plasma T levels or change them in any way (about 850 ng/100 ml throughout). The findings suggest that P implants may be preferable to weekly MPA injections in the treatment of male sex offenders because they require less patient compliance and may not have MPA's troubling side effects. PMID- 9226344 TI - Effects of a single intraseptal injection of NGF on spatial learning in the water maze. AB - Male Sprague-Dawley rats given electrolytic lesions of the septum followed by a single intraseptal injection of 5 microg of NGF were trained on a water maze task that assessed their ability to learn the location of a visible platform and the location of platform when it was submerged. Rats with damage to the septum acquired the visible platform version of the task but were significantly impaired in locating the submerged platform. Administration of NGF, however, produced an intermediate ameliorative effect on the measure of latency to find the hidden platform during these trials. In order to determine the relative strength of the place and cue responses learned during the visible and hidden platform training trials, a probe trial was given on the final test day in which the visible platform was moved to a new location. Control rats swam either to the new platform location or the old platform location indicating the use of both a place and cue response. However, both rats with septal damage alone and rats with septal lesions treated with NGF swam directly to the new platform location indicating the relative strength of the cue response. These results support previous findings indicating that a single injection of NGF can produce improvements on a cognitive task, but it may not be doing so by restoring lost spatial functions following septohippocampal damage. PMID- 9226345 TI - Separation as a new animal model for self-induced weight loss. AB - Animal models for weight loss are generally either stress mediated or following diet restriction (DR) schedules. We investigated weight loss in mice subjected to activity stress (ACT), DR schedules of 40, 60 and 100% of daily requirements, and propose a new model for animal weight loss based upon separation. Mice were separated (SEP) from each other by perspex partitioning for 23 h per day, with free access to food for one hour. No significant differences in weight loss were found between the ACT, SEP and 40% groups. However, mean food intake in the 40% group was half that of the ACT group (p < 0.001) and significantly less (p < 0.01) than the SEP group, which consumed amounts equivalent to 65% of daily requirement. Separation of mice increases metabolic demands and may be used as a new, easily performed, animal model for weight loss. PMID- 9226346 TI - 24-hour recordings of blood pressure, heart rate and behavioural activity in rabbits by radio-telemetry: effects of feeding and hypertension. AB - We used radio-telemetry to measure 24-hour rhythms of systolic, diastolic and mean blood pressure, heart rate and behavioural activity in conscious rabbits, which were maintained under normal day/night rhythms and restricted feeding. Over three consecutive days, all variables showed little change between day-period and night-period, except for a pronounced rise in the afternoon, coinciding with the presentation of pellet food. Mean blood pressure increased during this period from baseline values between 78-82 mm Hg to a peak of 89-91 mm Hg. At the same time heart rate rose from baseline values of 147-161 b/min to a peak of 206-234 b/min and behavioural activity scores rose from 11-31 counts/h to a peak of 52-81 counts/h. Changing the time at which pellet food was presented to the rabbits from the early afternoon to the early morning, caused a complete and immediate shift of the peak of blood pressure and heart rate to the morning period. Chronic intravenous infusion of angiotensin II caused a significant increase in blood pressure (24-hour average: 80 +/- 1 vs. 114 +/- 7 mm Hg) but did not alter basal heart rate or behavioural activity. The increase in heart rate and blood pressure seen with food presentation was attenuated with angiotensin II infusion. These data show that in rabbits diurnal changes in blood pressure, heart rate and activity were determined to a large extent by timed feeding. In addition, in rabbits with angiotensin-induced hypertension the food-induced changes in blood pressure and heart rate were blunted. PMID- 9226347 TI - Patterns of body temperature, activity, and reproductive behavior in a tropical murid rodent, Arvicanthis niloticus. AB - Nile grass rats (Arvicanthis niloticus), are murid rodents from tropical Africa that exhibit diurnal patterns of wheel-running. In the present paper we describe the temporal organization of several other behaviors in these animals, as well as daily rhythms in their body temperature. In the first experiment, we characterized rhythms of gross motor activity and core body temperature in four adult females implanted with telemetry transmitters and kept on a 12:12 light:dark (LD) cycle. In all animals body temperature and gross motor activity were clearly diurnal, with peaks often occurring around dawn and dusk. In the second experiment we recorded the times of mating and parturition in eight mating couples housed in a 12:12 LD cycle. We monitored animals 24 h a day using a time lapse video recording system, beginning when males and females were paired, and ending after the birth of the second litter and the associated post-partum copulation. Mating almost always began just before the lights came on, and parturition generally occurred in an "anticrepuscular" pattern, outside of the periods around dawn and dusk. Thus, these animals exhibit an interesting mosaic of temporal adaptations, with some crepuscular tendencies expressed within a predominantly diurnal pattern. PMID- 9226348 TI - Activity measures in rhesus monkeys on long-term calorie restriction. AB - Calorie restriction (CR), undernutrition without malnutrition, extends the mean and maximal lifespan of several ecologically diverse species. Rodents on CR demonstrate increased activity measured as spontaneous locomotion, wheel running, open field behavior or movement. Activity measures were recorded from 19 male rhesus monkeys (Macaca mulatta) as either controls (C) which were fed a nutritious diet to approximate ad libitum levels, or as experimentals (E) which were fed 30% less than age- and weight-matched controls. Within each diet group, some monkeys (n = 10) began CR at 2.3 years of age (range 2.2-2.4 yrs, J Group) while another group (n = 9) began CR at approximately 4.6 years of age (range 4 5.25, A group). Beginning about 6 years after initiation of the study, behavioral activity was measured via ultrasonic motion detectors and recorded on videotape. Diurnal and circadian activity was clearly discernible. Peaks in activity were associated with mealtime and colony husbandry. Compared to Group A, Group J monkeys exhibited higher overall activity as measured by sensors, and also significantly more circling. Compared to AC monkeys, group AE monkeys demonstrated higher rates of gross motor behavior, pacing, stereotypies and grooming. The increases in motor activity observed in one group of monkeys were consistent with results obtained from rodent studies of CR and aging. CR did not significantly inhibit or negatively influence the display of behavior of rhesus monkeys in the laboratory environment. We report here, for the first time, increases in activity due to CR in a model other than the rodent. PMID- 9226349 TI - Daily exercise reduces fat, protein and body mass in male but not female rats. AB - This study was designed to compare the estimated energy balance, linear growth (body and bone lengths) and body composition (all components including body mass, total body water, fat, protein and ash) response to daily spontaneous running (DSR) in young male and female rats. We tested the hypothesis that due to gender differences in energy efficiency, DSR would reduce linear growth and body composition more in male rats. Fourteen male and sixteen female weanling Sprague Dawley rats were randomly assigned to either a sedentary (SED) control (male 7, female 8) or DSR (male 7, female 8) group. The DSR rats were allowed to run spontaneously in running wheels while SED rats remained in standard rat cages for 9 weeks. Body mass, running distance and food intake were measured over the nine week period. Subsequently, chemical analysis was performed to measure carcass content of water, protein, fat and ash. Linear growth was assessed by measures of body and bone lengths. The estimated energy balance of the DSR rats was computed and compared between genders. Estimated energy balance was significantly more negative in females than males due to significantly greater DSR distance. Body and bone lengths were similar among the SED and DSR female and SED and DSR male rats. However, whole body mass, fat mass and protein mass were significantly lower only in DSR males. These results demonstrate that DSR reduced body mass, body fat and protein mass in male rats but not in female rats despite a more negative estimated energy balance in female rats. These findings suggest that females are better protected from an energy deficit due to DSR. Possible mechanisms include gender-specific hormonal responses. PMID- 9226350 TI - Oil enriched diets and behavioral parameters in rats' recovery from early undernutrition. AB - The effect of oil enriched diets on sexual and exploratory behavior has been studied in male rats undernourished in utero and during lactation. At 20 days of age, for a period of 4 months, these animals were fed with 3 different diets: standard diet, standard diet enriched with 7% soybean oil, and standard diet enriched with 7% olive oil. A control group eating standard diet was also studied. Sexual behavior, open-field and nocturnal spontaneous locomotor activity were studied at 16-20 weeks of age. Undernutrition produced decreased body weight, and the experimental diets were not effective in growth recovery. Undernourished animals eating oil-enriched diets displayed lower nocturnal spontaneous locomotor activity, less time spent in central squares, and a lower number of rearing episodes than undernourished animals eating the standard diet; the number of peripheral squares entered was, however, significantly increased. Experimental groups eating oil diets showed a statistically significant increase in the number of ejaculating males and in the total number of ejaculations compared with experimental rats eating standard diet. These findings suggest that early undernutrition produces permanent behavioral alterations in male rats, but that the sexual behavior deficit could be reversed by feeding our oil-enriched diets from weaning. The diet enriched with soybean oil seems more effective in the rehabilitation of sexual behavior. PMID- 9226351 TI - Opioid-receptor subtype agonist-induced enhancements of sucrose intake are dependent upon sucrose concentration. AB - Selective mu ([D-Ala2, N-Me-Phe4, Gly-ol5]-enkephalin (DAMGO)), delta1 ([D-Pen2, D-Pen5]-enkephalin (DPDPE)), delta2 ([D-Ala2, Glu4]-Deltorphin (Delt II)), kappa1 (U50488H) and kappa3 (naloxone benzoylhydrazone (NalBzOH)) opioid agonists each stimulate food intake in rats. Whereas studies with selective opioid antagonists implicate mu and kappa1 receptors in the mediation of sucrose intake, studies with selective opioid agonists implicate mu and delta receptors in the mediation of saccharin intake. The present study determined if specific delta1, delta2, kappa1, kappa3 and mu opioid-receptor subtype agonists produced similar alterations in sucrose intake as a function of sucrose concentration (0.5%, 2.5%, 10%) across a 1-h time-course. Each of these agonists significantly increased sucrose intake with variations in pattern, magnitude, and consistency as a function of sucrose concentration. Whereas the mu opioid agonist, DAMGO, and the delta1 opioid agonist, DPDPE, each enhanced sucrose intake at higher (2.5%, 10%), but not lower (0.5%), concentrations, the delta2 opioid agonist, Delt II, increased sucrose intake at lower (0.5%, 2.5%), but not higher (10%), concentrations. Kappa opioid agonists produced less consistent effects. The kappa1 opioid agonist, U50488H, increased sucrose intake at high (10%) concentrations and decreased sucrose intake at low (0.5%) concentrations, and the kappa3 opioid agonist, NalBzOH, inconsistently increased sucrose intake at the 0.5% (20 microg) and 10% (1 microg) concentrations. Thus, these data further implicate mu, delta1, and delta2 opioid mediation of palatable intake, particularly of its orosensory characteristics. PMID- 9226353 TI - Postnatal testicular secretions partially restore the disturbances in reproductive activity caused by prenatal hormonal manipulation. AB - Male rats that prenatally had been exposed to an antiestrogen, nitromifene citrate (CI 628), showed evidence of impaired defeminization and masculinization in adulthood, suggesting a role of endogenous estrogen for the sexual differentiation of the male. The present study was undertaken to investigate a possible role of postnatal testicular secretions for the above behavioral effects. Male rats were exposed prenatally to CI 628 (1 mg/rat) or saline, and castrated on Day 0, Day 10, or Day 90 after birth. After treatment with gonadal hormones in adulthood, the males were tested for feminine and masculine sexual behavior and for sexual orientation, both when sexually naive and after they had acquired sexual experience. The following conclusions were drawn: 1. Permanent deficits of lordotic behavior were observed in all experimental groups, suggesting the importance of prenatal estrogen for the defeminization process. 2. Hop/darting and ear wiggling behaviors were enhanced in Day-0 and Day-10 castrates, and blocked in the Day-90 castrates. The restitution of these behaviors to normal levels in Day-90 castrates suggests that, in addition to prenatal estrogen, postnatal testicular secretions also are involved in the behavioral defeminization process. 3. Prenatal estrogen contributes to masculinization as evidenced by the impaired ejaculatory behavior observed in all experimental groups. 4. Male-typical sexual orientation toward the female seems to be facilitated by prenatal estrogen. Both the masculinization and the defeminization of male-typical sexual orientation toward a female were impaired by castration at birth and at Day 10 in the experimental animals, but a full restoration of the sexual orientation toward females was seen in Day-90 castrates, suggesting the restorative role of postnatal testicular secretions for these processes. PMID- 9226352 TI - Morphological and metabolic changes associated with large differences in daily food intake in crossed-intestines rats. AB - Twenty-two inbred male Lewis rats were made into parabiotic pairs and 7 pairs had a further operation in which the small intestines of the 2 rats were connected so that one rat continually lost food into the upper small intestine and bloodstream of its partner. As a result, these rats showed large and sustained changes in daily food intake with one rat (A) in each pair eating more than twice as much as its partner (B) for the rest of their lives. Measurements of plasma levels of glucose, insulin, and glucagon did not vary directly with daily food intake, but integrated plasma lactate values were lower in rats that ate more (A) and higher in rats that ate less (B). At sacrifice, the rats that ate more were found to have less fat with reduced fat cell size but the same cell number in both retroperitoneal and epididymal fat pads. Measurements of the rate and pattern of glucose metabolism in retroperitoneal fat cells with or without insulin stimulation were similar across groups. Rates of lipolysis with and without epinephrine did not differ among groups. Lipoprotein lipase varied directly with fat cell size and indirectly with daily food intake. These studies show that daily food intake varies directly with fat cell size and inversely with plasma lactate and retroperitoneal lipoprotein lipase levels. PMID- 9226354 TI - Deafferentation of the olfactory bulbs of male rats reduces erection to remote cues from females. AB - If male rats are paired with a receptive, soliciting female rat in a setting that prevents physical contact, but permits auditory, visual, and olfactory contact, some will have erections of the penis--noncontact erections (NCE). Surgical deafferentation of the olfactory bulbs from all the known chemosensory systems of the nasal septum renders rats anosmic, decreases sexual performance in copulation tests with females, and substantially reduces the frequency of NCE. Thus, NCE appear to be primarily regulated though the olfactory perception of volatile chemosensory cues from receptive females. PMID- 9226355 TI - Scanning behavior of rats during eating under stressful noise. AB - Behavioral ecological theories postulate that threatening environments should increase eating speed and vigilance during feeding. In the present experiment, eating speed and scanning behavior during eating were measured in 36 rats in 5 consecutive test sessions under stressful noise (95 dB white noise, n = 18) and control conditions (60 dB, n = 18) after the animals had been habituated to the test environment. Intense noise induced an increase of scanning rate and eating speed. These effects are similar to those reported for novel and light environments. PMID- 9226356 TI - The effect of dietary fat on salivary habituation and satiation. AB - This study was designed to explore the effect of dietary fat and carbohydrate on oral habituation. Forty women (18-21 years old) were randomized to 1 of 4 yogurt conditions that varied levels of dietary fat and carbohydrate. Subjects consumed 0.4 g of yogurt per pound per trial until they indicated fullness in up to 15 trials, with salivation, number of trials to fullness, and ratings of hedonics, appetite, and fullness measured. Subjects in the high-fat conditions demonstrated a significantly faster rate of salivary habituation than subjects in the low-fat conditions, and consumed less volume of yogurt but consumed more calories. The rate of habituation was not influenced by carbohydrate content of the yogurt. These findings suggest that oral habituation in humans is sensitive to macronutrient content of food. PMID- 9226357 TI - A novel chronic and detachable indwelling jugular catheterization procedure for mice. AB - The purpose of this study was to describe and demonstrate the usefulness of a chronic, detachable, indwelling jugular catheterization apparatus in mice that can be applied to acute or chronic I.V. drug administration in freely moving, unrestrained mice. The application of this procedure to the study of abused drugs is particularly advantageous, because the commonly employed intraperitoneal (I.P.), S.C., and per os (P.O.) routes of administration fail to mimic the near complete and instantaneous bioavailability of drugs abused by I.V. injection or inhalation (e.g., cocaine). Compared to current I.V. administration methods, the detachable catheter system presented in this paper is relatively easy to construct, simple to use, and appears to remain patent for an extended period of time. In addition, the utility of this procedure is greatly increased because the subject does not have to remain permanently attached to a complicated tether system. Thus, the test subject can be infused with a drug, detached from the catheter, and then undergo some behavioral test that would otherwise be impossible to undergo with a tethered system. For the purposes of demonstrating catheter patency and the importance of an I.V. route of administration, a conditioned place preference (CPP) paradigm was used to evaluate the reinforcing efficacy of cocaine. PMID- 9226358 TI - Elemental and configural learning and the perception of odorant mixtures by the spiny lobster Panulirus argus. AB - The present study used a conditioning assay to investigate if the type of learning task that spiny lobsters (Panulirus argus) were required to perform influenced the way that they perceived odorant mixtures. Mixtures were composed of 2 food-related compounds (adenosine-5'-monophosphate, betaine, or L-glutamate) at concentrations that produced the same duration of searching behavior in unconditioned animals. Aversive conditioning of search behavior coupled with generalization testing was used to evaluate perceptual similarity between related mixtures. When animals were conditioned to stop searching to a binary mixture AX, they did not generalize significantly from this mixture to either of its components (A or X), or to a binary mixture containing one novel component (AY). However, when lobsters were conditioned to avoid AX but to continue responding to AY, they generalized between AX and X and between AY and Y. The results support the hypothesis that altering the salience of a mixture's components by giving them different reinforcement contingencies changed the way that the mixtures were perceived. As a result of such conditioning, animals perceived the mixture's components as separate elements, rather than as a configuration, and, as a consequence, animals generalized between binary mixtures and their most salient or predictive components. PMID- 9226360 TI - The effects of mixing on behavior and circadian parameters of salivary cortisol in pigs. AB - The present study describes an experiment that was carried out to study the effects of mixing pigs once at 25 kg, preceded by transportation for 1.5 h, on the behavior and the circadian rhythmicity of salivary cortisol. The frequency of agonistic interactions was higher for mixed pigs. This was not only the case immediately after mixing, when pigs started to fight to establish a new social rank (p < 0.05), but also 5 to 6 weeks later; still more headknocks and bites towards other pigs were seen at that time among mixed pigs (p < 0.01). However, neither the basal cortisol concentration, assessed as the MESOR of the circadian rhythm of salivary cortisol, nor the amplitude of that rhythm was different between the groups. PMID- 9226359 TI - Selective activation of CCK-B receptors does not induce sleep and does not affect EEG slow-wave activity and brain temperature in rats. AB - Systemic injections of cholecystokinin octapeptide sulfate ester (CCK-8-SE) elicit various behavioral and autonomic responses, such as increases in nonrapid eye-movement sleep (NREMS) and hypothermia. There are two CCK receptors; both CCK A and CCK-B receptors are stimulated by CCK-8-SE. The relative importance of the CCK-A and CCK-B receptors in the somnogenic and hypothermic effects of CCK-8-SE is not well understood. In the present experiments, we studied the effects of the selective activation of CCK-B receptors by CCK tetrapeptide (CCK-4) or nonsulfated CCK-8 (CCK-8-NS) on sleep and brain temperature (Tbr). Rats were injected intraperitoneally with saline on the control day and with CCK-8-NS (10, 50, or 250 microg/kg) or CCK-4 (10, 50, or 250 microg/kg) on the test day 5-10 min before dark onset. Electroencephalogram, electromyogram, and Tbr were recorded for 12 h. None of the treatments affected sleep or Tbr significantly, with the exception of 10 microg/kg CCK-4, which transiently decreased the amount of NREMS, and 10 microg/kg CCK-8-NS, which slightly increased REMS. These results suggest that the activation of CCK-B receptors by systemic injection of CCK-4 or CCK-8-NS is not sufficient to elicit increased NREMS and hypothermia in rats. PMID- 9226361 TI - The influence of food on pain perception in healthy human volunteers. AB - The aim of this study was to investigate if food could reduce pain perception in a group of 16 healthy human volunteers (8 male and 8 female), and to explore the differential effects of macronutrient composition on the response to cold-induced pain. All subjects underwent the cold pressor test (CPT) on 3 occasions in a counterbalanced order, before and after administration of isoenergetic high-fat low-carbohydrate (CHO) and high-CHO low-fat meals, and when no meal was given. The CPT was carried out 4 times on each test day, once before the meal, and 0.5, 1.5, and 2.5 h after the meal, and at the equivalent times on the day when no food was given. Radial pulse and blood pressure measurements and visual analogue scales of mood/emotional state were carried out before and after each CPT. Mean pain scores were significantly reduced following both meals compared with the no food condition. The maximum reduction in pain occurred 1.5 h after ingestion, and a significantly greater effect was exerted by the high-fat low-CHO meal compared with the high-CHO low-fat meal. These results demonstrate that food, particularly when rich in fat, significantly reduces the pain induced by the cold pressor stimulus in healthy human subjects. PMID- 9226362 TI - A comparison of the ability of 8-9-year-old children and adults to detect taste stimuli. AB - Conflicting data exist in the literature regarding the maturity of the human sense of taste during childhood and if gender influences gustatory development. To investigate these 2 questions, taste detection thresholds for the 4 common tastants sucrose, sodium chloride, citric acid, and caffeine were established for 61 young adults and 68 children aged 8-9 years old, using a paired-comparison forced-choice procedure. No significant differences were found between the mean thresholds of women and men, or between those of female children and adults. In contrast, male children had significantly higher thresholds for all 4 tastants than adult females, for all tastants except caffeine than adult men, and for sucrose and sodium chloride than female children. It is concluded that the taste sensitivity of 8-9-year-old males, although well developed, has not fully matured, and that taste sensitivity is not affected by gender in young adults. PMID- 9226363 TI - Pain-related disability and effects of chronic morphine in the adjuvant-induced arthritis model of chronic pain. AB - Functional disability has been identified as one of the most important aspects of chronic pain, yet modeling pain-related disability has received little attention. Adjuvant-induced arthritis was induced, and one group of arthritic rats was implanted with SC 75-mg morphine pellets 1 week postadjuvant, and reimplanted every 2 weeks thereafter. The results confirm that the rodent adjuvant-induced arthritis model of severe chronic pain can be used to model pain-related disability: spontaneous activity levels and ambulatory function were reduced in arthritic rats and they exhibited substantial weight loss. The results of the present study suggest that the operant delayed nonmatching-to-position task can be used as a measure of pain-related disability, which may be especially relevant to the effects of chronic pain on performance in a work setting. The delayed nonmatching-to-position operant bar-pressing task is an "apical" test that is sensitive to deficits across a wide range of behavioral functions: motor ability, attention, motivation, learning, and memory, and arthritic rats were severely impaired in this task. In addition, analgesic treatments that impair functional abilities in normal healthy rats may actually improve the performance of rats exhibiting pain-related disability. Previous work demonstrated that acute morphine injections of only 4 mg/kg impaired performance in the delayed matching to-position task. The results of the present study demonstrate that chronic morphine attenuates the degree of pain-related disability exhibited by arthritic rats in the test of ambulatory function and the delayed nonmatching-to-position bar-pressing test. These results demonstrate that novel analgesic treatments can be screened preclinically, both with respect to their direct analgesic effects, and with respect to their ability to reduce pain-related disability. PMID- 9226364 TI - Early postnatal treatment with transforming growth factor alpha does not alter nonreproductive behavior. AB - Estrogen acting during the critical developmental period has been postulated to defeminize and possibly masculinize male sexual behavior. Transforming growth factor alpha (TGF alpha) also may be involved, because this growth factor, at least partly, mediates the mitotic effects of estrogen on target tissues. Male transgenic mice overexpressing TGF alpha have elevated serum estradiol (E2) levels and they exhibit feminization of many nonreproductive actions, suggesting that either TGF alpha and/or E2, or both, participate in the control of some nonreproductive behavior. Male and female CD-1 mice were treated with 4 microg of recombinant human TGF alpha or 2-4 microg E2 during the first 3 days of life. Although early TGF alpha treatment accelerates physical development and influences the growth of the uterus and mammary gland, it failed to have any effect on behavior, either in male or female mice. Early E2 treatment significantly lengthened immobility time in the swim test and reduced voluntary alcohol intake among the male mice. No changes in locomotor activity or aggressive behavior were noted. The expression of TGF alpha mRNA in the brainstem of adult male mice was not altered following neonatal TGF alpha or E2 treatment. However, neonatal exposure to TGF alpha caused a moderate elevation in TGF alpha mRNA expression in the female brainstem. Our results indicate that in male, but not in female mice, an excess of E2 during early life affects some nonreproductive behavior. Furthermore, early treatment with recombinant human TGF alpha does not alter nonreproductive behavior in mice. PMID- 9226365 TI - Evolution of delta activity within the nonREM sleep episode: a biphasic hypothesis. AB - The time course of delta activity within nonREM (NREM) episodes is measured for 24 healthy subjects with normal REM latencies. The first two NREM episodes in particular, show two very clearly separated peaks for about 35% of the subjects. Another 25% show two less well separated peaks. These double peak patterns are also prevalent in the literature, but there has been a tendency to dismiss them as a skipped REM effect. They are, however, still evident even when the data are averaged over the 24 subjects, indicating a systematic phenomenon. These averaged data are well fitted by an analytic function given by the sum of two consecutive overlapping Gaussian curves. The well-behaved residuals also, are an indication that a biphasic model of this kind is statistically appropriate. The model proposed is simple, with parameters related to physiological phenomena, and it suggests that there may be an underlying process with delta waves emanating from two separate signal sources. Recent neurophysiological findings suggest that delta oscillations are generated both in the thalamus and in the cortex and show that excessive synchronization of slow oscillations may lead to seizures. Hence the speculation that the biphasic process may emanate from cortical and thalamic sources and be protective in the sense that it permits smaller delta amplitudes at each source while retaining the integral delta energy necessary to satisfy sleep pressure. It is significant that the two peaks are most evident in the first two NREM episodes where delta power is high. PMID- 9226366 TI - Suggestive evidence for a schizophrenia susceptibility locus on chromosome 6q and a confirmation in an independent series of pedigrees. AB - We have investigated whether there is a locus on chromosome 6 that confers an increased susceptibility to schizophrenia using a two-stage approach and nonparametric linkage analysis. Allele sharing identical by descent (IBD) and multipoint maximum likelihood score (MLS) statistics were employed. Results from two tested data sets, a first data set, or genome scanning data set, and a second replication data set, show excess allele sharing for multiple markers in 6q, a chromosomal region not previously reported as linked to schizophrenia. In our genome scanning data set, excess allele sharing was found for markers on 6q13 q26. The greatest allele sharing was at interval 6q21-q22.3 at marker D6S416 (IBD percentage 69; P = 0.00024). The multipoint MLS values were greater than 2.4 in the 11.4-cM interval delimited by D6S301 and D6S303, with a maximum value of 3.06 close to D6S278 and of 3.05 at D6S454/D6S423. We did not confirm, however, the previously described linkage in 6p, when tested in the systematic genome scanning data set. The replication data set also showed excess allele sharing in chromosomal area 6q13-q26, which overlapped with the aforementioned positive linkage area of the genome scanning data set. The highest sharing of the second data set was at D6S424 (IBD percentage 64; P = 0.0004), D6S283 (IBD percentage 62; P = 0.0009), and D6S423 (IBD percentage 63; P = 0.0009). Multipoint MLS analysis yielded MLS values greater than 1 in an area of about 35 cM, which overlaps with the MLS multipoint area of linkage from the genome scanning data set. The multipoint MLS at the D6S454/D6S423 locus was 2.05. In the second data set, the maximum multipoint MLS was located about 10 cM centromeric from the maximum of the genome scanning data set, at the interval D6S424-D6S275 (2.35). Our results provide very suggestive evidence for a susceptibility locus for schizophrenia in chromosome 6q from two independent data sets. PMID- 9226367 TI - Localization of somatostatin receptor genes on mouse chromosomes 2, 11, 12, 15, and 17: correlation with growth QTLs. AB - A major role of the peptide hormone somatostatin is inhibition of growth hormone secretion. The effects of somatostatin are mediated through five distinct G protein-coupled receptors, each of which is expressed in the pituitary gland and other tissues. Allelic variation in the five somatostatin receptor genes (Smstr) could contribute to growth rate and overall body size. To evaluate this hypothesis we determined the chromosomal location of the Smstr genes. Restriction fragment length polymorphisms and single-strand conformational polymorphisms were used to follow the segregation of each gene in interspecific mouse backcrosses. Smstr1 through Smstr5 were localized to mouse chromosomes 12, 11, 15, 2, and 17, respectively. None of the Smstr genes colocalized with single gene mutations that affect growth. However, growth is a quantitative trait influenced by many genes and by the environment. Strains of mice selected for high and low growth have been exploited to identify chromosomal regions that modestly influence growth (J. Cheverud et a1., 1996, Genetics 142: 1305-1319). Several Smstr genes map within these regions, suggesting that they be considered candidate genes for these quantitative trait loci. PMID- 9226368 TI - The mouse gene for the inducible G-protein-coupled receptor edg-1. AB - Edg-1, an immediate-early gene induced during the in vitro differentiation of human endothelial cells, encodes a G-protein-coupled receptor (GPR) that signals via the G(i)/mitogen-activated protein kinase (MAP kinase) pathway (Lee, M.-J., Evans, M., and Hla, T. (1996) J. Biol. Chem. 271, 11272-11279). It is a prototypical member of the subfamily of "orphan" receptors that are expressed in the cardiovascular and nervous systems. In this report, the mouse edg-1 gene was cloned and sequenced, and its expression patterns were defined. The edg-1 transcript was expressed in a wide variety of adult tissues including the brain, lung, liver, heart, and spleen. However, during embryogenesis, the edg-1 mRNA was induced late in development (after Embryonic Day 15.5) at centers of ossification. As a first step toward understanding the molecular basis of tissue specific and inducible expression, the mouse edg-1 gene and its promoter were characterized. The mouse edg-1 gene is composed of two exons and is 4.9 kb in length. The second exon is large and contains the entire coding region and the 3' untranslated region. The edg-1 promoter is TATA-less and contains GC-rich elements, and transcription initiation occurs from a single start site. The 5' flanking region of the promoter contains several enhancer elements. However, the activity of the 5'-flanking region was suppressed by the repressor activity within the first intron. These data provide the basis for the further characterization of the regulation of the orphan GPR edg-1. PMID- 9226369 TI - High-resolution physical mapping of a 6.7-Mb YAC contig spanning a region critical for the monosomy 21 phenotype in 21q21.3-q22.1. AB - Deletion of genes from the chromosome 21 region between APP and SOD1 is a potential cause of some of the major phenotypic features of monosomy 21 patients. Fine physical mapping helps identify potential candidate genes. After selecting nonchimeric YACs by FISH analysis, we determined their marker contents by PCR and hybridization studies. Fifteen YACs were chosen and mapped by restriction enzyme analysis and labeling of end fragments. We localized 55 markers, including 31 STSs, 10 YAC ends, and 4 NotI linking clones, along a 6.7-Mb contig. This map facilitates transcriptional analysis of this region and construction of ready-to sequence contigs. Furthermore, FISH mapping of two patients with partial monosomy 21 using YAC and cosmid clones allowed us to define more accurately the telomeric border of the critical region between markers S226 and S213. PMID- 9226370 TI - Isolation and identification of the human homolog of a new p53-binding protein, Mdmx. AB - We recently reported the identification of a mouse cDNA encoding a new p53 associating protein that we called Mdmx because of its structural similarity to Mdm2, a well-known p53-binding protein. Here we report the isolation of a cDNA encoding the human homolog of Mdmx. The ORF of the cDNA encodes a protein of 490 amino acids, 90% similar to mouse Mdmx. The homology between Mdmx and Mdm2 is most prominent in the p53-binding domain and the putative metal-binding domains. The Mdmx protein, which, based on SDS-PAGE, has a MW of 80 kDa, can bind p53 in vitro. The human MDMX gene is transcribed in all tissues tested, with high levels in thymus. By fluorescence in situ hybridization analysis we mapped the mouse mdmx gene to chromosome 1 (region F-G) and the human MDMX gene to chromosome 1q32. PMID- 9226371 TI - Cloning of a novel human neural cell adhesion molecule gene (NCAM2) that maps to chromosome region 21q21 and is potentially involved in Down syndrome. AB - To contribute to the development of the transcription map of human chromosome 21 (HC21), we have used exon trapping to identify portions of HC21 genes. One trapped exon showed strong homology with members of the neural cell adhesion molecule (NCAM) family of genes from different species. We subsequently cloned the complete coding sequence from a human fetal brain cDNA library and determined its nucleotide sequence and predicted amino acid sequence. The predicted polypeptide of this novel NCAM2 gene contains 837 amino acids and shows 62% similarity to the NCAM homologs. It contains five immunoglobulin-like domains, two fibronectin type III domains, a transmembrane domain and a cytoplasmic domain. The gene is expressed most strongly in human adult and fetal brain. Using somatic cell hybrids, we mapped NCAM2 to 21q21, between markers D21S18 and D21S282. Radiation hybrid mapping localized this novel gene between polymorphic markers D21S1914 and D21S265. NCAMs are members of the immunoglobulin superfamily and are essential in the formation and maintenance of tissue structure. To date there are no candidate human disorders on HC21 that could be associated with mutations in NCAM2. In addition, the role of NCAM2 in the pathophysiology of Down syndrome is unknown. However, it is a good candidate for involvement in certain Down syndrome phenotypes because a slight overexpression of NCAMs increases many fold the homotypic adhesion properties of cells. PMID- 9226372 TI - Identification of endogenous retroviral sequences based on modular organization: proviral structure at the SSAV1 locus. AB - The current genome sequencing projects reveal megabases of unknown genomic sequences. About 1% of these sequences can be expected to be of retroviral origin. These are often severely deleted or mutated. Therefore, identification of the retroviral origin of these sequences can be very difficult due to the absence of convincing overall sequence similarity. There are also many copies of solo LTRs (long terminal repeats) distributed throughout genomic sequences. LTR and envelope sequences in general are among the most divergent parts of the retroviral genome and thus especially hard to detect in mutated endogenous sequences. We took advantage of the fact that these retroviral sections contain short highly conserved sequence regions providing retroviral hallmarks even after loss of overall similarity. We defined several sequence elements and peptide motifs within LTR and Env sequences and used these elements to construct models for LTRs and Env proteins of mammalian C-type retroviruses. We then used this strategy to identify successfully the hitherto missing LTRs and an env-like region in the S71 human retroviral sequence. Our approach provides a new strategy for identifying remotely related retroviral sequences in genomic DNA (especially human DNA), of potential significance for the interpretation of genomic sequences obtained from the current large-scale sequencing projects. PMID- 9226373 TI - Genetic and physical maps of the stargazer locus on mouse chromosome 15. AB - The stargazer mouse mutation causes absence seizures that are more prolonged and frequent than any other petit mal mouse model. Stargazer mice also have an ataxic gait and vestibular problems, including a distinctive head-tossing motion. From the genotyping of a large intersubspecific cross, a panel of 53 recombinant DNAs between D15Mit29 and D15Mit2 has been assembled, and a fine genetic map of the stargazer region has been constructed on mouse Chromosome 15. The stargazer locus has been mapped between D15Mit30 and the parvalbumin gene, and six candidate genes have been excluded by genetic linkage analysis. A physical contig of YACs, BACs, and P1s stretching 1.1 Mb from D15Mit30 to the somatostatin receptor 3 gene is reported, and the DNA interval including the stargazer locus has been narrowed to 150 kb. PMID- 9226374 TI - A novel human prostate-specific, androgen-regulated homeobox gene (NKX3.1) that maps to 8p21, a region frequently deleted in prostate cancer. AB - We have isolated a prostate-specific gene (NKX3.1) in humans that is homologous to the Drosophila NK homeobox gene family. Northern blot analyses indicate that this gene is expressed at high levels in adult prostate and at a much lower level in testis, but is expressed little or not at all in several other tissues. In an androgen-dependent prostate carcinoma line, LNCaP, NKX3.1 mRNA is expressed at a basal level that was increased markedly upon androgen stimulation; the NKX3.1 mRNA was undetectable in several other human tumor cell lines including two androgen-independent prostate carcinoma lines. The NKX3.1 gene maps to chromosome band 8p21, a region frequently reported to undergo a loss of heterozygosity associated with tissue dedifferentiation and loss of androgen responsiveness during the progression of prostate cancer. Based on these data we propose that NKX3.1 is a candidate gene for playing a role in the opposing processes of androgen-driven differentiation of prostatic tissue and loss of that differentiation during the progression of prostate cancer. PMID- 9226376 TI - An integrated physical and genetic map spanning chromosome band 10q24. AB - Chromosome band 10q24 is rich in genes involved in development, tumorigenesis, neurological disorders, hormone metabolism, and environmentally induced disease susceptibility. We have constructed an STS-based integrated physical and genetic map of 10q24 derived from the CEPH-Genethon mega-YAC contig data for this region. This map consists of 42 fluorescence in situ hybridization-mapped overlapping CEPH mega-YACs spanning approximately 15 Mb to which 49 STS markers have been assigned, including 24 Genethon CA repeat genetic markers, 10 known gene loci from the 10q24 region (IFI56, IDE, PDE6C, RBP4, CYP2C, CD39, DNTT, GOT1, WNT8B, and PAX2) and 11 additional expressed sequences of unknown function. PMID- 9226375 TI - The structural integrity of ROR alpha isoforms is mutated in staggerer mice: cerebellar coexpression of ROR alpha1 and ROR alpha4. AB - The recessive mouse mutation staggerer (sg) disturbs the normal development of cerebellar Purkinje cells and affects certain functions of the immune system. To identify the causative gene, we constructed high-resolution genetic and physical maps of the staggerer locus on mouse chromosome 9. The transcription unit of the orphan nuclear receptor ROR alpha was identified in the critical interval. Our mutational analysis confirms a recent report that the sg phenotype may be caused by a genomic deletion in the common coding region of the ROR alpha isoforms. Of the four different isoforms of the ROR alpha gene that are generated by a combination of alternative promoter usage and exon splicing that differ in their DNA-binding properties, isoforms ROR alpha1 and ROR alpha4 are specifically coexpressed in the murine cerebellum and human cerebellum. Thus, at least two isoforms of the murine ROR alpha gene are affected by the genomic deletion associated with the staggerer phenotype. Our finding of cerebellum-specific coregulation suggests that distinct sets of target genes regulated by the ROR alpha1 and ROR alpha4 isoforms are required for Purkinje cell development. PMID- 9226377 TI - Genomic structure and assignment of the RhoH/TTF small GTPase gene (ARHH) to 4p13 by in situ hybridization. AB - The RhoH/TTF (ARHH) gene encodes a new member of the Ras superfamily of small GTPases. The gene was identified by fusion to the BCL6/LAZ3 oncogene in an initially described t(3;4)(q27;p11) translocation in a non-Hodgkin's lymphoma cell line. The predicted amino acid sequence of the RhoH/TTF gene product includes Rho-like GTPase structural motifs. The RhoH/TTF gene is restrictively expressed in hematopoietic cells and tissues. Mutations in the human RAS genes have been shown previously to be tumorigenic; in the search for a potential implication of the RhoH/TTF gene in hemopoietic malignancies, we established its genomic structure. The RhoH/TTF gene spans 35 kb and contains two exons, with the second bearing the entire amino-acid-coding region. Chromosomal mapping, by FISH experiments, places the RhoH/TTF gene on the short arm of chromosome 4, band p13. PMID- 9226378 TI - Yeast artificial chromosome transfer into human renal carcinoma cells by spheroplast fusion. AB - Successful transfer of yeast artificial chromosomes (YACs) into human cells has been described in only a single study. We here report on the evaluation of YAC transfer strategies into a human renal cell carcinoma cell line by yeast spheroplast fusion and cationic lipids. While the latter approach proved inefficient, significant numbers of clones containing both vector arms were obtained by spheroplast fusion. FISH analyses on such clones revealed the presence of YAC integration and the co-localization of both vector arms with insert sequences. These data demonstrate that under certain experimental conditions efficient YAC transfer into human cells by spheroplast fusion is possible and may be useful for the cloning of human disease-related genes. PMID- 9226379 TI - Cloning and mapping of the MEIS1 gene, the human homolog of a murine leukemogenic gene. AB - The mammalian homeobox genes encode a family of transcription factors that are important in a wide range of cellular processes, including hematopoiesis. Aberrant expression of some homeobox genes is known to be oncogenic. We report the cloning and initial characterization of a human homeobox gene, MEIS1, identified in a survey of homeobox genes expressed in the human fetal liver. The complete cDNA sequence shows that MEIS1 is likely to be the human homolog of Meis1, a mouse gene that is known to be activated in myeloid leukemia by retroviral insertion. We found that the MEIS1 gene is expressed at low levels in normal immunohematopoietic tissues, including the fetal liver. However, consistent with its possible role in myeloid leukemogenesis, MEIS1 was expressed in a subset of myeloid leukemia cell lines, with the highest expression seen in those with a megakaryocytic-erythroid phenotype. The gene is also expressed at high levels in the cerebellum. The gene is located on human chromosome region 2p13-p14 and contains the previously identified anonymous markers D2S134 and NIB1519. Whether this gene, which is leukemogenic in mice, also plays a leukemogenic role in humans will require further study. PMID- 9226380 TI - Isolation of a novel human homologue of the gene coding for echinoderm microtubule-associated protein (EMAP) from the Usher syndrome type 1a locus at 14q32. AB - Usher syndrome type 1 (USH1) is an autosomal recessive, genetically heterogeneous disorder causing severe congenital deafness, retinitis pigmentosa, and vestibular dysfunction. The USHla locus located on 14q32 has been linked to the genetic markers D14S250 and D14S78. Using D14S250 and D14S78, we have isolated two nonchimeric YACs, 878g10 and 844g2, and a single BAC (135i20) and PAC (194e17) clone and have arranged them into a contig spanning over the D14S250 and D14S78 markers. The analysis of the YACs, BAC, and PAC revealed that the physical distance between D14S250 and D14S78 is less than 25 kb. Iterative cDNA library screening initiated with the EST 219670 found in the vicinity of the D14S78 marker yielded a cDNA contig. The nucleotide sequence of the cDNA encodes a protein of 717 amino acids in length, showing a high level of homology to the Echinoderm 77-kDa microtubule-associated protein (EMAP). The human homologue of Echinoderm microtubule-associated protein defines a novel human gene. We propose that the human EMAP is a strong candidate for the USH1a gene based on its genomic location and the proposed function of the protein. PMID- 9226381 TI - Physical mapping of human myosin-IXB (MYO9B), the human orthologue of the rat myosin myr 5, to chromosome 19p13.1. PMID- 9226382 TI - JAK3 maps to human chromosome 19p12 within a cluster of proto-oncogenes and transcription factors. PMID- 9226383 TI - Localization of the Rab escort protein-2 (REP2) and inositol 1,4,5-trisphosphate 3-kinase (ITPKB) genes to mouse chromosome 1 by in situ hybridization and precision of the syntenic regions between mouse and human 1q42-q44. PMID- 9226384 TI - The influence of endodontic treatment upon periodontal wound healing. AB - The interrelationship between periodontal and endodontic disease has aroused much speculation, confusion, and controversy. Pulpal and periodontal problems are responsible for more than 50% of tooth mortality today. Diagnosis is often difficult since these diseases have been studied primarily as separate entities. The toxic substances of the pulp may initiate periodontal defects through canal ramifications and patent dentinal tubules, thus impairing wound healing in regenerative procedures. Although no studies exist addressing the direct effect of pulpal infection on the outcome of guided tissue regeneration (GTR) procedures, several studies do indicate that pulpal status may play a significant role toward the end results of GTR. This review article discusses the potential influence of endodontic treatment on the long-term outcomes of GTR. Potential pathways between the pulp and periodontal ligament, which may be responsible for the failure of the regeneration of new periodontal attachment apparatus, are explored. Examination and review of the clinical and research findings in the literature relating to perio-endo lesions are made to demonstrate that a negative influence may exist between GTR outcomes and the status of the pulp. PMID- 9226385 TI - Tetracycline demineralization of dentin: the effects of concentration and application time. AB - The current investigation was initiated to study the effect concentration and application time has on the rate of tetracycline demineralization of dentin. Buccal and lingual surfaces of extracted bovine molars were ground to a smooth flat dentin surface using wetted silicon carbide discs. Standardized depressions were made in the dentin surface with a #909-055 diamond round wheel. Fresh tetracycline HCl (TTC-HCl) (Flavine Int. Inc.) solutions, i.e., 0, 25, 50, 75, 100, 125 and 150 mg/ml were prepared. A 30% citric acid solution was used as a positive control. The pH of each solution was recorded. 7 microl of each solution were pipetted into a depression and remained undisturbed for 1, 3, or 5 min. At the end of each application time period a fresh #3 cotton pellet was placed in the depression, once every 20 s for 1 min, to soak up the solution. The 3 pellets were placed in a 2.00 ml of 18 M omega H2O sample. As a measure of the rate of demineralization, the parts per million calcium (ppm Ca++) found in each sample were determined using atomic absorption spectrophotometry. Two-way analysis of variance was used to determine effects of TTC-HCI concentration and time on the rate of demineralization. No significant differences were found in the mean ppm Ca++ released at 1-, 3- and 5-min application times for 0, 25, or 50 mg/ml TTC. No significant differences were found in the mean ppm Ca++ released (i) between 3 and 5-min application times for 75, 100, 125 and 150 mg/ml TTC-HCl solutions and (ii) between 75, 100, 125 and 150 mg/ml TTC-HCl solutions within either the 3- or 5-min application times. The mean ppm Ca++ released at 3- and 5-min application times for 75, 100, 125 and 150 mg/ml TTC-HCI solutions were all significantly greater than the respective readings at the 1-min application time. The mean ppm Ca++ recorded for the 30% citric acid solution for all 3 application times were 3 to 5.5 x greater than the highest mean ppm Ca++ recording for TTC-HCl. The results of this study show that a 3-min application time of 75 mg/ml TTC-HCl solution is equally as effective at demineralizing dentin as is higher concentrations and/or longer application times, but was far less effective than a 30% citric acid solution. PMID- 9226386 TI - Treatment of 3rd molar-induced periodontal defects with guided tissue regeneration. AB - Recent reports provide evidence of increased attachment levels when using guided tissue regeneration (GTR) techniques for the treatment of periodontal defects. Periodontal defects frequently occur at the distal aspect of mandibular 2nd molars which are next to mesioangular impacted 3rd molars that have oral communication. The purpose of this study was to determine whether the use of GTR can enhance probing attachment levels (PALs) following extraction of mesioangular impacted third molars. 12 patients with bilateral soft tissue impacted mandibular 3rd molars entered this split mouth study. After extractions, the previously exposed distal root surface of the 2nd molars were debrided. The defects on the randomly selected experimental sites were covered with expanded polytetraflouro ethylene (e-PTFE) membrane and the tissue was replaced to cover the membrane. Membranes were removed after 6 weeks. Control sites were treated identically except no membrane was placed. GI, P1I, PD, PAL and BOP records were obtained at 0, 3 and 6 months. The use of barrier material did not provide statistically significant differences in PAL when comparing experimental versus control sites. Nevertheless, PAL gain was consistently greater at 3 and 6 months when GTR techniques were used in sites with deep impactions. PMID- 9226387 TI - Clinical response to local delivery of tetracycline in relation to overall and local periodontal conditions. AB - The purpose of this study was to determine the clinical response to local delivery of tetracycline in relation to clinical and microbiological conditions of the other teeth. 4 deep pockets were monitored in 19 subjects with multiple deep periodontal lesions and high counts of P. gingivalis. In 9 patients (LT) only 2 of the selected lesions were treated by placement of tetracycline fibers (Actisite), while the rest of the dentition was left untreated. In the other 10 patients, all teeth were supragingivally scaled and then treated by application of polymeric tetracycline HCl containing fibers, the whole dentition was subject to full mouth scaling and root planing, and the patients rinsed with 0.2% chlorhexidine (FT). A significant reduction in mean PPD was observed in all treated sites after two months. This reduction was maintained over the following 4 months. The magnitude of the effect was significantly greater in the FT group (1.74 mm) than in the LT group (0.88 mm). The mean attachment level changes were similar after 2 months in locally and fully treated subjects. A tendency of relapse was noted for treated sites in LT patients from month 2 to 6. A level of statistical significance was not reached for this effect. Data from measurements recorded at 6 sites around all teeth in the full mouth treated patients were analyzed using multiple linear regression. This analysis showed local changes in PPD and AL were significantly and strongly correlated with the baseline value of the respective parameter at the same site. In addition, more pocket depth reduction was noted if a site was not bleeding on probing at 6 months, if the location of a site was not approximal and if the tooth was not a second molar. Sites located on second molars showed also less AL gain than sites located on other teeth. Smokers showed significantly less reduction in PPD and significantly less AL gain. Furthermore, if subjects had a high % of pockets deeper than 4 mm at baseline they showed significantly less attachment gain. PMID- 9226388 TI - Occurrence of invading bacteria in radicular dentin of periodontally diseased teeth: microbiological findings. AB - Bacterial invasion in roots of periodontally diseased teeth, which has been recently documented using cultural and microscopic techniques, may be important in the pathogenesis of periodontal disease. The purpose of this investigation was to determine the occurrence and the species of invading bacteria in radicular dentin of periodontally diseased teeth. Samples were taken from the middle layer of radicular dentin of 26 periodontally diseased teeth. 14 healthy teeth were used as controls. Dentin samples were cultured anaerobically. The chosen methodology allowed the determination of the numbers of bacteria present in both deeper and outer part of dentinal tubules, and the bacterial concentration in dentin samples, expressed as colony forming units per mg of tissue (CFU/mg). Invading bacteria was detected in 14 (53.8%) samples from periodontally diseased teeth. The bacterial concentration ranged from 831.84 to 11971.3 CFU/mg (mean+/ standard deviation: 3043.15+/-2763.13). Micro-organisms identified included putative periodontal pathogens such as Prevotella intermedia, Porphyromonas gingivalis, Fusobacterium nucleatum, Bacteroides forsythus, Peptostreptococcus micros and Streptococcus intermedius. These findings suggest that radicular dentin could act as bacterial reservoir from which periodontal pathogens can recolonize treated periodontal pockets, contributing to the failure of therapy and recurrence of disease. PMID- 9226389 TI - Effect of delmopinol hydrochloride mouthrinse on plaque formation and gingivitis in "rapid" and "slow" plaque formers. AB - The aim of the present study was to investigate differences in the plaque and gingivitis inhibiting effect of delmopinol rinsing between "rapid" and "slow" plaque formers. 23 subjects (12 "rapid" and 11 "slow" plaque formers) were selected from 71 healthy young adults. The selection was based on the plaque index on the buccal surfaces of all premolars and 1st molars after 3-days without plaque control. The 23 subjects were randomly assigned into 3 groups with different mouthrinses, i.e., 0.1% delmopinol, 0.2% delmopinol, and placebo. The study was double-blind with parallel design between the "rapid" and "slow" plaque formers and cross-over design between 2 active periods and a placebo period. Each rinsing period lasted for 5 days. During the 3 test periods, the subjects refrained from all oral hygiene and rinsed 2x daily with either one of the 3 solutions. Gingival crevicular fluid (GCF) was collected from buccal surfaces of upper canines and premolars and bleeding on probing (BOP) recorded at 6 sites around each tooth before and after each test period. Plaque assessment, including plaque index (PI) and standardized color slides for planimetric analyses obtained from the canines and premolars, were only recorded after each test period. Results showed that the mean PI and planimetry values for both the "rapid" and "slow" plaque formers were lower than the placebo, for either the 0.1% or the 0.2% delmopinol mouthrinse. The differences between the" rapid" and "slow" plaque formers were not statistically significant. There was a small reduction in BOP in both groups for the delmopinol periods, as against a slight increase in the placebo period; the difference between the placebo group and the 2 groups of plaque formers was not statistically significant (p>0.6 for both 0.1% and 0.2% delmopinol). Results suggested that both 0.1% and 0.2% delmopinol reduce plaque formation and gingivitis to a similar extent in subjects with extreme rates of plaque formation. PMID- 9226390 TI - A study on factors associated with pathologic tooth migration. AB - The purpose of this cross-sectional epidemiological study was to determine the prevalence of pathologic tooth migration (PTM) among periodontal patients and to investigate the relationship and degree of association between PTM and the following factors: bone loss, tooth loss, gingival inflammation according to the gingival index, age, lingual interposition, parafunctions and oral habits. 852 periodontal patients (36.7% male, 63.3% female) whose ages ranged from 19 to 72 years (mean 42.5 +/- 9.9) were studied. PTM was defined as the presence of a developing diastema in the upper anterior sextant, which was not present in the past or already existed but increased. Statistical analysis was performed using the Wald test and the Mantel-Haenszel test. Estimates odds ratio were also calculated to assess increased PTM probability as a function of a single variable, or a combination of several. PTM prevalence of 55.8% was found, and it was statistically associated with bone loss (p<0.001), tooth loss (p<0.001) and gingival inflammation (p<0.001), while no association was observed with the remaining variables. The odds ratio indicated that PTM probability increased between 2.95 to 7.97 times as bone loss increased. For tooth loss this probability increased 2.76 times when no tooth loss was compared to 4 or more teeth lost. Likewise the probability increased 2.23 times when the gingival index was above 2. According to the single effect as well as the combined effect of these 3 main factors, it was concluded that: (1) no single factor by itself is clearly associated with PTM: (2) the factor mainly related to PTM is bone loss, followed by tooth loss and gingival inflammation: (3) as bone loss increases, the association of additional factors with PTM, such as tooth loss and gingival inflammation, increases. PMID- 9226391 TI - Effectiveness of subgingival instrumentation with power-driven instruments in the hands of experienced and inexperienced operators. A study on manikins. AB - Power instrumentation of periodontally-diseased root surfaces is gaining in significance as an alternative to conventional curette methods. In an experimental study employing manikins with simulated bone loss, we investigated whether inexperienced and experienced operators were able to achieve greater therapeutic success with power-driven devices than with hand instruments in subgingival scaling. 10 dentists experienced in periodontal treatment and 10 inexperienced dentists instrumented 7 teeth in the upper jaw, which had been covered with artificial deposits. Hand instruments, the Perioplaner system, a sonic and an ultrasonic scaler were used. The time required for treatment was measured and the % of residual deposits was calculated by means of image processing techniques. Weight loss was also determined for the teeth that were scaled with the hand instruments and the Perioplaner system. Experienced operators left significantly less % of residual deposits on the teeth (18+/-7.6%) than the inexperienced (27+/-8.4%), regardless of the type of instrument selected. Both experienced and inexperienced operators left the smallest amounts of residual deposits with hand instruments (13+/-9.8%/24+/-9.5%). Both treatment groups removed more hard tooth structure with hand instruments than with the Perioplaner system (53+/-48mg versus 47+/-25.9 mg). Experienced operators needed somewhat more time for debridement than unexperienced. Use of the sonic/ultrasonic device required somewhat less time than hand instrumentation. Inexperienced operators are, however, unable to improve their treatment results by using the power-driven instruments included in the study. PMID- 9226392 TI - Periodontal disease related to diabetic status. A pilot study of the response to periodontal therapy in type 1 diabetes. AB - Variation in the periodontal health status and the response to oral hygiene education, scaling and root planing were studied in 36 subjects with type-1 diabetes mellitus (DM) and in 10 non-diabetic control subjects. The age range of the subjects was 24-36 years. The diabetic group was divided into 3 subgroups based on the levels of glycosylated hemoglobin (HbAlc) over a 3 year period and the presence of diabetic complications as follows: (D1) subjects with good metabolic control and no complications (n=13), (D2) subjects with varying metabolic control with/without retinopathy (n=15) and (D3) subjects with severe diabetes, i.e., with poor long-term control and/or multiple complications (n= 8). Clinical measurements (plaque, subgingival calculus, probing pocket depth, bleeding after probing and clinical attachment level) were performed at the baseline and 4 weeks and 6 and 12 months after periodontal therapy. The between group comparisons were made using the Student t-test and ANOVA. Based on the plaque scores, the oral hygiene status was similar in all groups during the whole study. No statistically-significant differences in the periodontal health status could be found between the diabetic group as a whole and the non-diabetic controls at any examination. The level of periodontal health of the diabetics with good control and no complications (D1) and those with moderate control with/without retinopathy (D2) was on the same level with that seen in the non diabetic controls. Our findings of the significantly higher extent of al > or =2 mm at the baseline and the fast recurrence of pd > or =4 mm during the longitudinal study in diabetic subjects with poor metabolic control and/or multiple complications (D3) indicate increased periodontal breakdown as a complication of DM in these subjects. To be able to assess the periodontal prognosis and the need for periodontal therapy on an individual basis,the clinical practitioner should be well aware of the diabetic status of his/her patients. PMID- 9226393 TI - Clinical evaluation of a bioabsorbable regenerative material in mandibular class II furcation therapy. AB - 30 periodontally compromised adult subjects with mandibular buccal class II furcation defects were recruited for this study. All selected defects were treated according to the biological principles of guided tissue regeneration. The subjects were randomly assigned to 2 parallel groups. The test group (n=15) received a bioabsorbable polyglycolic-polylactic membrane (PGA/PLA group); the control group (n=15) received a non-resorbable expanded polytetrafluoroethylene membrane (ePTFE group). After initial therapy, baseline measurements were recorded including plaque index, gingival index, vertical and horizontal probing depths, clinical attachment level and depth of the recession. Recall visits were made at 1, 2, 4, 6, 8, 12, and 24 weeks. At 12 months, all baseline clinical parameters were again measured. The data analysis did not demonstrate a significant difference between the 2 groups. The vertical probing depth and attachment level changes were statistically significant in each group. The postoperative recession was 0.6 mm in the ePTFE group (p<0.05) and 0.8 mm (p<0.05) in the PGA/PLA group. Compared to the initial measurements, the mean changes in horizontal probing depth were 2.7 mm and 2.5 mm (p<0.001), corresponding to mean reductions of 41.5% and 40.9% for the ePTFE and the PGA/PLA groups respectively. The results of this study suggest that 12 months after initial surgery, similar clinical improvements can be obtained in GTR therapy of buccal class II furcation lesions, regardless of whether bioabsorbable PGA/PLA membranes or non-resorbable ePTFE membranes are used. PMID- 9226394 TI - Pathways mediating CRF-induced inhibition of gastric emptying in rats. AB - The corticotrophin-releasing factor (CRF) is shown to be released during stress suggesting that CRF has a physiological role in the mediation of central nervous system (CNS) response to stress, including an inhibitory effect on gastric emptying. In the present study, we have examined the pathways by which intracerebroventricularly (i.c.v.) administered CRF and central CRF activation during stress alter the gastric emptying rate of saline (0.14 M), acid (50 mM), peptone (4.5%) and peptone after preload. The emptying rates of all these test meals were significantly (p < 0.05-0.001) delayed with increasing doses of i.c.v. CRF (0.001, 0.003, 0.01, 0.1, 0.3 and 1 nmol/10 microl), when compared with their i.c.v. saline-treated controls. The 1-nmol dose of CRF inhibited the emptying of acid, peptone and peptone after a preload by 43.8%, 64.1% and 81.1%, respectively. Twenty-minute swim stress delayed gastric emptying rate of saline, acid and peptone solutions significantly (p < 0.001) and the CRF receptor antagonist, alpha-helical CRF (8 nmol/10 microl, i.c.v.), applied before the swim stress, abolished the inhibitory effect of stress on the emptying rate of these solutions. Acute intragastric administration of capsaicin (2 mg/rat) and systemic capsaicin (125 mg kg(-1)) treatment facilitated the gastric emptying rate of acid, peptone and peptone after preload significantly, almost abolishing the inhibitory effect of central CRF (p < 0.001). However, either capsaicin treatment had no effect on stress-induced inhibition of the gastric emptying of none of the solutions, except peptone after a preload. Our findings demonstrate that the gastric inhibitory response induced by swimming as a stress-producing stimulus is mediated by the endogenous release of CRF. They also suggest that CRF exerts its CNS actions on the gastrointestinal tract via vago-vagal, capsaicin-sensitive pathways, probably involving the central cholecystokinin (CCK) mechanisms. PMID- 9226395 TI - Transforming growth factor alpha-immunoreactivity in neural tissues of the rat stomach. AB - We report TGF alpha immunoreactivity in neurons of the myenteric plexus and in nerve fibers in the muscle and submucosal layers of the rat stomach. Association of TGF alpha staining nerve fibers to vessels and smooth muscle cells gives morphological evidence that EGF/TGF alpha's actions to increase mucosal blood flow and gastric motility may be mediated by TGF alpha derived from neural structures. These data suggest that TGF alpha plays a role in the neural control of the gastric function. PMID- 9226396 TI - CGRP potentiates excitatory transmission to the circular muscle of guinea-pig colon. AB - We aimed to assess whether calcitonin gene-related peptide (CGRP) can modulate the release of tachykinins which are the main nonadrenergic noncholinergic (NANC) excitatory transmitters to the circular muscle of the guinea-pig proximal colon. In organ bath experiments, electrical field stimulation (EFS) in the presence of atropine (1 microM) and guanethidine (3 microM) evoked twitch phasic NANC contractions which were abolished by the combined administration of tachykinin NK1 and NK2 receptor antagonists. Human alphaCGRP (CGRP, 1-100 nM) produced a concentration-dependent potentiation of the amplitude of the NANC contractions induced by EFS while salmon calcitonin (up to 1 microM) had no effect. The potentiating effect of CGRP was unaffected by in vitro capsaicin pretreatment (10 microM for 15 min), peptidase inhibitors (captopril, bestatin and thiorphan, 1 microM each), apamin (0.3 microM) plus L-nitroarginine (L-NOARG, 100 microM) and by the CGRP1 receptor antagonist, the C-terminal fragment CGRP(8-37) (1 microM). The NK2 receptor antagonist MEN 10627 which, when administered alone, had only a partial inhibitory effect on the amplitude of NANC twitches, concentration dependently (10 nM-1 microM) inhibited the potentiating effect of CGRP. CGRP (1 100 nM) produced a concentration-dependent potentiation of the atropine-sensitive cholinergic contractions evoked by EFS in the presence of guanethidine and of tachykinin NK1 and NK2 receptor antagonists. Similar to the effect of CGRP, application of capsaicin (0.1-1 microM) potentiated the amplitude of the NANC contraction to EFS, an effect undergoing complete desensitization upon a second application of the drug. CGRP (0.1 microM) did not affect the contractile action of a submaximally effective concentration of neurokinin A (2 nM) while it inhibited that induced by substance P (2 nM). In sucrose gap, single pulse EFS in the presence of atropine (1 microM) and guanethidine (3 microM) induced an inhibitory junction potential (i.j.p.) and a small excitatory junction potential (e.j.p.). CGRP (0.1 microM) produced membrane hyperpolarization and relaxation without affecting i.j.p. amplitude but concomitantly increased the e.j.p. amplitude to induce a contraction in correspondence to each electrical pulse. In the presence of the NK1 receptor antagonist, GR 82334 (3 microM), the membrane hyperpolarization and relaxation produced by CGRP and the EFS-evoked i.j.p. were unaffected, while the potentiating effect of CGRP on the EFS-evoked NANC e.j.p. and the corresponding contraction were abolished. We conclude that, in addition to the previously characterized direct smooth muscle relaxant action via CGRP1 receptors (Maggi et al. Regulatory Peptides 61, 27-36, 1996), CGRP also induces a remarkable potentiation of excitatory neurotransmission to the circular muscle of the guinea-pig colon via CGRP2 receptors. The latter effect, documented in this study, is evidenced on both the atropine-sensitive and the atropine-resistant (tachykinin-mediated) components of excitatory transmission: this effect does not involve mediator(s) release from capsaicin-sensitive primary afferent nerves, nor inhibition of peptide degradation or modulation of NANC inhibitory transmission. PMID- 9226397 TI - Idazoxan and the effect of intracerebroventricular oxytocin or vasopressin on sodium intake of sodium-depleted rats. AB - The alpha2-adrenergic agonist clonidine and the neuropeptide oxytocin, inhibit sodium intake when injected intracerebroventricularly (i.c.v.). The present work investigates whether (1) vasopressin also inhibits sodium intake when injected i.c.v., and (2) the effect of oxytocin and of vasopressin on sodium intake is affected by i.c.v. injection of idazoxan, an alpha2-adrenergic antagonist. Clonidine (30 nmol), oxytocin (40, 80 nmol) and vasopressin (40, 80 nmol) were injected i.c.v. 20 min prior to a 1.5% NaCl appetite test, in rats depleted of sodium for 24 h by a combination of a single s.c. injection of furosemide (10 mg/rat) and removal of ambient sodium. Every dose of clonidine, oxytocin and vasopressin inhibited the 1.5% NaCl intake. Seizures were observed with the higher dose of vasopressin, but not with either dose of oxytocin. The effect of i.c.v. injection of clonidine (30 nmol), oxytocin (80 nmol) or vasopressin (40 nmol) was partially inhibited by prior i.c.v. injection of idazoxan (160, 320 nmol). The results suggest that the inhibition of 1.5% NaCl intake induced by i.c.v. injection of neuropeptides in sodium-depleted rats depends, in part, on the activation of central alpha2-adrenoceptors. PMID- 9226398 TI - Somatostatin modulates the function of Kupffer cells. AB - Recent studies have shown that somatostatin modulates lymphocyte and peritoneal macrophage function, but the effects of somatostatin on hepatic macrophages (Kupffer cells) are not clearly defined. In the present study, hepatic macrophages obtained from male rats were treated in vitro with somatostatin or octreotide and their effects on the release of nitrite, tumor necrosis factor (TNF), and hydrogen peroxide (H2O2) determined. At concentrations of 10(-14) M to 10(-12) M, or greater than 10(-10) M, somatostatin suppressed nitrite release by Kupffer cells. At concentrations of less than 10(-9) M or greater than 10(-9) M, octreotide inhibited nitrite release by Kupffer cells. Kupffer cells treated with 10(-10) M to 10(-14) M or greater than 10(-8) M of somatostatin released significantly less amounts of TNF than did the untreated controls. TNF release by Kupffer cells treated with 10(-15) M to 10(-5) M of octreotide was significantly inhibited as compared to that of untreated controls. Kupffer cells treated with 10(-14) M to 10(-11) M and 10(-9) M to 10(-8) M of somatostatin released more H2O2 than did the untreated controls. The amount of H2O2 released by noncirrhotic Kupffer cells treated with 10(-6) M or 10(-5) M of somatostatin was less than that of controls. These findings demonstrate that somatostatin and octreotide modulate the release of nitric oxide, TNF, H2O be Kupffer cells depending on the concentration of hormones used. PMID- 9226399 TI - GIP(6-30amide) contains the high affinity binding region of GIP and is a potent inhibitor of GIP1-42 action in vitro. AB - GIP (Glucose-dependent Insulinotropic Polypeptide) is an important regulator of insulin secretion. The effects of truncated forms of the peptide, GIP(10-30), GIP(6-30amide) and GIP(7-30), on binding of 125I-GIP(1-42) to GIP receptors in transfected CHO-KI cells, and on cyclic AMP responses to GIP(1-42), have been studied with a view to defining further the receptor binding region of GIP, and to establish whether such truncated peptides exhibit agonist or antagonist activity. All three peptides were found to be receptor antagonists, however GIP(6 30amide) exhibited receptor binding affinity equivalent to that of GIP(1-42) in competitive binding studies (IC50 = 3.08+/-0.57 nM). GIP(6-30amide) inhibited GIP(1-42)-induced cAMP production by 58% at a concentration of 100 nM, whereas GIP(10-30) and GIP(7-30), inhibited only in the microM range. GIP(6-30amide) therefore contains the high affinity binding region of GIP and is a potent inhibitor of GIP(1-42) action in vitro. PMID- 9226400 TI - Distribution and functional effects of neuropeptide Y on equine ureteral smooth muscle and resistance arteries. AB - The distribution of neuropeptide Y (NPY)-immunoreactive (IR) nerves, as well as the functional effects of NPY and the Y1- and Y2-receptor agonists, [Leu31,Pro34]NPY and NPY(13-36), respectively, have been investigated in vitro in both visceral and arterial smooth muscle of the horse intravesical ureter. NPY-IR nerve fibres were widely distributed along the entire length of the ureter, although the intravesical part was the most richly innervated region, and the only one where NPY-IR ganglion cells were found. NPY (10(-7) M) did not affect either basal tone or spontaneous rhythmic contractions of the isolated intravesical ureter, but significantly enhanced the increases in both tone and frequency of phasic activity elicited by noradrenaline (10(-6) and 10(-5) M). The Y1-receptor agonist, [Leu31,Pro34]NPY (10(-7) and 10(-6) M) did not significantly alter either ureteral basal tone or the contractile activity induced by noradrenaline, whereas the Y2-receptor agonist, NPY(13-36) (10(-7) M), mimicked the potentiating effect of NPY on noradrenaline responses. In ureteral resistance arteries (effective lumen diameters of 130-300 microm), NPY (10(-10) to 10(-7) M) elicited concentration-dependent contractions, which were inversely correlated with the arterial lumen diameter. Submaximal concentrations of NPY (10(-8) M) significantly increased the sensitivity of ureteral arteries to noradrenaline. [Leu31,Pro34]NPY (10(-10) to 10(-7) M), but not NPY(13-36), induced a contractile effect of similar magnitude and potency as those of NPY, and also potentiated noradrenaline responses. The present results demonstrate a rich NPY-innervation in the intravesical ureter and reveal functional effects of the peptide enhancing motor activity in both ureteral and arterial smooth muscles, although the receptors mediating such effects seem to be different. Thus, NPY potentiates the phasic contractions and tone elicited by noradrenaline through Y2-receptors, whereas it both contracts and potentiates noradrenaline vasoconstriction in ureteral arteries via Y1-receptors. PMID- 9226401 TI - Adenosine A1 receptors mediate hypoxia-induced inhibition of electrically evoked transmitter release from rat striatal slices. AB - We have examined the role of adenosine in mediating effects of mild hypoxia on electrically evoked transmitter release. Rat striatal slices, preincubated with [3H]dopamine and [14C]choline, were superfused continuously and stimulated electrically. Before and during the second stimulation, some slices were superfused with Krebs' solution with lowered oxygen. This mild hypoxia caused a significant increase of the electrically evoked outflow of endogenous adenosine, hypoxanthine and inosine into the superfusion buffer, whereas electrically evoked release of [3H]dopamine and [14C]acetylcholine was significantly decreased. The addition of 8-cyclopentyl-1,3-dipropylxanthine, a selective adenosine A1 receptor antagonist, blocked the hypoxia-induced inhibitory effect on the evoked release of these two transmitters in a concentration-dependent manner. In summary, the results show that reduction of the oxygen supply to striatal slices results in an increased release of endogenous adenosine, which, by acting on adenosine A1 receptors, decreases the electrically evoked release of dopamine and acetylcholine. PMID- 9226402 TI - Serotonergic modulation of central respiratory activity in the neonatal mouse: an in vitro study. AB - In order to determine whether the serotonergic modulation of the central respiratory activity previously reported in neonatal rats occurs in species other than the rat, we performed identical in vitro experiments on the neonatal mouse to those performed on the neonatal rat. The effects of adding serotonin (5 hydroxytryptamine, 5-HT) and related agents to the superfusate suggested that the respiratory rhythm generator undergoes an excitatory modulation via medullary 5 HT1A receptors. Upon applying the drugs to the spinal cord alone, 5-HT was found to have a dual effect on phrenic motoneuron firing: (i) a facilitatory effect mediated by 5-HT2A receptors and (ii) a depressive effect on their inspiratory discharge mediated by non-5-HT1A, non-5-HT2A, non-5-HT3 receptors, possibly of the 5-HT1B subtype. It was therefore concluded that serotonin modulates the neonatal central respiratory activity in mice as well as in rats, and that similar 5-HT receptor subtypes are involved in this process in both species. PMID- 9226404 TI - Chronic zolpidem treatment alters GABA(A) receptor mRNA levels in the rat cortex. AB - The effect of chronic zolpidem treatment on the steady-state levels of gamma aminobutyric acidA alpha1-6, beta1-3 and gamma1-3 subunit mRNAs in rat cortex has been investigated. Male Sprague-Dawley rats were injected once daily, for 7 or 14 days, with 15 mg/kg of zolpidem in sesame oil vehicle. The levels of the alpha4 and beta1 subunit mRNAs were significantly increased after 7 days of treatment and the level of alpha1 subunit mRNA was significantly decreased after 14 days of treatment, as determined by solution hybridization. These results are compared to the previously determined effects of an equivalent schedule of treatment with diazepam. PMID- 9226403 TI - Ethanol as a general anesthetic: actions in spinal cord. AB - Ethanol, usually studied in relation to intoxication, is also capable of producing general anesthesia. The most common standard of anesthetic potency is the concentration which produces immobility in response to a noxious stimulus. This concentration will be referred to as the anesthetic concentration. Immobilization is a spinal effect. Ethanol effects were studied in spinal cord from 2-7-day-old rats at concentrations which included the anesthetic concentration in both adult rats (97 mM) and 6-7-day-old rats (235 mM). At neonatal but not adult anesthetic concentrations, ethanol depressed monosynaptic reflex amplitude (mediated by glutamate AMPA receptors + compound action potential). At both neonatal and adult anesthetic concentrations ethanol reversibly depressed the population excitatory postsynaptic potential (pEPSP) (glutamate AMPA and NMDA receptors), the slow ventral root potential (NMDA + metabotropic receptors), and the dorsal root potential (GABA(A) receptors, via glutamate-excited interneurons). Effects were greater on NMDA receptor-mediated components than on AMPA-receptor-mediated components of the pEPSP and greater on NMDA than on metabotropic receptor-mediated components of the slow ventral root potential. The profile of ethanol effects on spinal cord resembles that of inhalation general anesthetics. The results show that both AMPA and NMDA receptor mediated transmission are sensitive to ethanol and that enhancement of GABAergic neurotransmission is overridden by depression of excitation to the interneurons. They provide no obvious explanation for ethanol's lower general anesthetic potency in the neonate. PMID- 9226405 TI - Actions of thienyl analogs of baclofen at GABA(B) receptors in rat neocortical slices. AB - In rat neocortical slices maintained in Mg2+-free Krebs medium, baclofen and its thienyl analogs, 4-amino-3-(5-chlorothien-2-yl)-butanoic acid (5h), 4-amino-3-(5 methylthien-2-yl)-butanoic acid (5d), 4-amino-3-(5-bromothien-2-yl)-butanoic acid (5f) and 4-amino-3-(thien-3-yl)-butanoic acid (5j) dose-dependently suppressed the spontaneous discharges, antagonised by the GABA(B) receptor antagonist 2 hydroxysaclofen (200 microM). Their relative potencies were baclofen > 5h > 5d > 5f > 5j. These heterocyclic analogs may prove useful as GABA(B) receptor agonists in functional studies. PMID- 9226406 TI - Aminoguanidine reverses aortic hyporeactivity to noradrenaline in portal vein ligated rats. AB - To evaluate the role of the inducible and endothelial constitutive nitric oxide synthase in vascular hyporeactivity to vasopressors in portal hypertension, in vitro experiments were performed on intact and endothelium-denuded isolated thoracic aortic rings from portal vein-ligated and sham-operated rats in control conditions, in the presence of aminoguanidine alone, considered to be a selective inhibitor of the inducible nitric oxide synthase, and of aminoguanidine and the nonselective nitric oxide synthase inhibitor N(G)-nitro-L-arginine. In control conditions, hyporeactivity to noradrenaline was observed in both rings with and without endothelium from portal hypertensive versus sham-operated rats. In the rings with endothelium, aminoguanidine reverted this hyporeactivity in portal hypertensive rats. N(G)-Nitro-L-arginine caused an additional shift to the left of the concentration-response curves to noradrenaline in portal hypertensive and a similar shift in sham-operated rats. In the endothelium-denuded rings, aminoguanidine caused no significant changes in portal hypertensive rats, whereas a significant shift to the right in the sham-operated rats was noted, however similar as the shift in the time controls not preincubated with aminoguanidine. No significant further changes were observed after preincubation with the two inhibitors. The endothelium-dependent relaxations to acetylcholine were attenuated in portal hypertensive versus sham-operated rats; addition of aminoguanidine shifted the relaxation curves to the left in portal hypertensive but not in sham-operated rats. These results provide indirect evidence for an increased activity of the inducible nitric oxide synthase in the intact aortic rings but not in the endothelium-denuded rings from portal vein-ligated rats, where other factors seem to be responsible for the observed hyporeactivity to noradrenaline. The endothelial constitutive nitric oxide synthase in rings from portal vein-ligated rats shows a reduced activity which is alleviated after inhibition of the inducible enzyme by aminoguanidine. PMID- 9226407 TI - Effects of adrenergic stimulation on sciatic nerve blood flow in diabetic and control rats. AB - Sciatic endoneurial blood flow is reduced in experimental diabetes. This study examined the possible involvement of noradrenergic mechanisms in this impairment. In anaesthetised rats (pentobarbitone sodium 50 mg/kg, diazepam 2 mg/kg), sciatic nerve laser Doppler flux and vascular resistance in diabetic rats (5-6 weeks) were lower (approximately 50%) and higher (approximately 42%), respectively, than that in age-matched control rats, indicating nerve ischaemia in the diabetic tissues. Tyramine (1 nmol), noradrenaline (0.001-1 nmol) and phenylephrine (0.01 10 nmol) produced significant increases of nerve vascular resistance in control rats. The responses to tyramine (1 nmol) were completely blocked by desipramine (10 nmol) and those to phenylephrine (10 nmol) were reversed by phentolamine (1 nmol). In streptozotocin-diabetic rats, responses to phenylephrine or noradrenaline were enhanced compared to control rats, but the enhancement failed to reach statistical significance. The findings demonstrate that adrenergic stimulation affects sciatic nerve endoneurial blood flow. PMID- 9226408 TI - Apamin-sensitive K+ channels involved in the inhibition of acetylcholine-induced contractions in lamb coronary small arteries. AB - In vitro experiments were designed to investigate the endothelial factors involved in modulation of the contractile response to acetylcholine in lamb coronary small arteries. Endothelial cell removal, and inhibitors of the L arginine/nitric oxide (NO) pathway increased basal tension and contractions in response to acetylcholine and abolished relaxations in response to the Ca2+ ionophore, 6S-[6alpha(2S*,3S*),8beta(R*),9beta,11alpha]-5-( methylamino)-2 [[3,9,11-trimethyl-8-[1-methyl-2-oxo-2-(1H-pyrrol-2-yl)et hyl]-1,7 dioxaspiro[5.5]-undec-2-yl]methyl]-4-benzoxazole carboxylic acid (A23187). N(G) Nitro-L-arginine enhanced acetylcholine-induced contractions in the absence, but not in the presence of the muscarinic M1 receptor antagonist, telenzepine. In contrast to glibenclamide and charybdotoxin, apamin enhanced the acetylcholine induced contractions and reduced the relaxations caused by A23187 and exogenously added NO. The combination of 1H-[1,2,4]oxadiazolo[4,3,-alpha]quinoxalin-1-one (ODQ) and apamin did not further increase the acetylcholine-induced contractions. These results indicate that muscarinic M1 receptor-released endothelial NO inhibits the contractile responses to acetylcholine in lamb coronary small arteries through activation of guanylate cyclase, followed by an increase in apamin-sensitive K+ conductance of the smooth muscle. PMID- 9226409 TI - Involvement of alpha1B-adrenoceptors in the positive inotropic effect of endogenous noradrenaline in rabbit myocardium. AB - We studied the alpha1-adrenoceptor subtypes mediating the positive inotropic effects of phenylephrine and noradrenaline as well as endogenous noradrenaline released by tyramine in rabbit papillary muscle. In the presence of propranolol, both phenylephrine and tyramine produced a positive inotropic effect in a concentration-dependent manner. WB4101 (N-[2-(2,6-dimethoxyphenoxy]ethyl]-2,3 dihydro-1,4-benzodioxin+ ++-2-methanamine) and chlorethylclonidine each antagonized the positive inotropic effect of phenylephrine. On the other hand, only chlorethylclonidine significantly blocked the positive inotropic effect of tyramine. However, the presence of both antagonists was needed to block the positive inotropic effect elicited by the exogenous addition of the low concentration of noradrenaline. These data suggest that after extensive blockade of beta-adrenoceptors the positive inotropic effects of phenylephrine and exogenous noradrenaline result from stimulation of the alpha1A- and alpha1B adrenoceptor subtypes, whereas that of endogenous noradrenaline is mediated via the alpha1B-adrenoceptor subtype. This could be explained by assuming that the alpha1B-adrenoceptor subtype population may be located on a space confronting more closely to the sympathetic nerve endings than the alpha1A-adrenoceptor subtype population. PMID- 9226410 TI - The ecto-ATPase inhibitor ARL 67156 enhances parasympathetic neurotransmission in the guinea-pig urinary bladder. AB - The influence of enzymatic degradation on the neurotransmitter actions of ATP was studied using the ecto-ATPase inhibitor 6-N,N-diethyl-D-beta,gamma dibromomethyleneATP (ARL 67156). Field stimulation of the parasympathetic nerves innervating guinea-pig urinary bladder muscle strips (1-8 Hz for 20 s) produced characteristic biphasic contractions, the peak magnitudes of which were significantly increased by 29-32% by ARL 67156 (100 microM). A similar degree of enhancement was seen in the presence of atropine (1 microM), consistent with ARL 67156 acting to enhance the action of neuronally released ATP. The effects of ARL 67156 reversed rapidly on washout of the drug. Contractions evoked by exogenous ATP (100 microM) were also potentiated by ARL 67156 (100 microM), but those to the stable analogue alpha,beta-methyleneATP (5 microM) were unaffected. ARL 67156 (100 microM) also enhanced contractions to exogenous acetylcholine (1 microM) and histamine (3 microM), but this potentiation was abolished by pyridoxalphosphate-6 azophenyl-2',4'-disulphonic acid (PPADS) (100 mciroM). It is concluded that when ATP acts as a neurotransmitter its postjunctional actions are attenuated by enzymatic degradation. ARL 67156 inhibits this breakdown. PMID- 9226411 TI - Effects of the nitric oxide-donor, GEA 3175, on guinea-pig airways. AB - This investigation characterized the smooth muscle relaxing effect of a novel nitric oxide (NO)-releasing substance, GEA 3175 (1,2,3,4-oxatriazolium, 3-(3 chloro-2-methylphenyl)-5-[[(4-methylphenyl)sulfonyl]amino], hydroxide inner salt) on guinea-pig trachea. GEA 3175 caused a concentration-dependent relaxation of tracheal smooth muscle precontracted with acetylcholine. This effect was reversed by both okadaic acid, an inhibitor of serine/threonine-specific phosphatases, and iberiotoxin, an inhibitor of Ca2+-activated K+ channels. Furthermore, GEA 3175 had a relaxation potency similar to that of the commonly used NO-donor, S-nitroso N-acetyl-penicillamine. On the contractile response provoked by electrical field stimulation, GEA 3175 induced a long-lasting relaxation which persisted even after repeated washing. The relaxing effect of GEA 3175 was associated with rises in guanosine 3':5'-cyclic monophosphate (cGMP). In time course studies, cGMP continued to increase with incubation time after stimulation with GEA 3175 and there was a significant elevation of cGMP even after washing. In contrast, incubation with S-nitroso-N-acetyl-penicillamine caused a transient rise in cGMP. The present investigation showed that GEA 3175 evokes long-lasting effects on contractile responses and cGMP levels in guinea-pig trachea. Our results indicate that the relaxing effect of GEA 3175 occurs through a mechanism involving phosphatases and iberiotoxin-sensitive K+ channels. PMID- 9226412 TI - Aceclofenac blocks prostaglandin E2 production following its intracellular conversion into cyclooxygenase inhibitors. AB - Aceclofenac, 2-[(2,6-dichlorophenyl) amino] phenylacetoxyacetic acid, is a novel non-steroidal anti-inflammatory drug. We investigated the effects of aceclofenac on prostaglandin E2 production by several kinds of human cells. Aceclofenac inhibited interleukin-1beta-induced prostaglandin E2 production by human rheumatoid synovial cells, but had no inhibitory effect on cyclooxygenase-1 or cyclooxygenase-2 activities by itself. We also observed that part of the aceclofenac was converted into diclofenac, the cyclooxygenase-1 and cyclooxygenase-2 inhibitor, when aceclofenac was incubated with human rheumatoid synovial cells. Aceclofenac was also converted into diclofenac and 4'-hydroxy diclofenac by human polymorphonuclear leukocytes and monocytes. 4'-Hydroxy diclofenac suppressed prostaglandin E2 production specifically by blocking cyclooxygenase-2 activity. These findings suggested that aceclofenac can be metabolized to cyclooxygenase inhibitors (diclofenac and/or 4'-hydroxy diclofenac) by these inflammatory cells. Although detailed examinations in non inflammatory cells remain to be studied, we concluded that aceclofenac is shown to be a new type of non-steroidal anti-inflammatory drug which is intracellulary converted into active metabolites that inhibit the prostaglandin E2 production. PMID- 9226413 TI - Differential effects of forskolin and 1,9-dideoxy-forskolin on nicotinic receptor and K+-induced responses in chromaffin cells. AB - The diterpene forskolin inhibits nicotine-evoked chromaffin cell Ca2+ influx, scinderin redistribution, F-actin disassembly and catecholamine secretion in a concentration-dependent (10-50 microM) fashion. On the other hand, forskolin showed weak inhibitory effects when the same responses were elicited by K+ induced depolarization. Similar concentrations of 1,9-dideoxy-forskolin, a forskolin analog which does not activate adenylate cyclase, blocked very effectively the responses evoked by either of the two stimuli. Patch-clamp (whole cell configuration) studies demonstrated that both diterpenes blocked fast and reversibly peak and total chromaffin cell nicotinic acetylcholine receptor currents, effects not mediated through adenylate cyclase activation. Moreover, both forskolin and 1,9-dideoxy-forskolin exhibited Ca2+ channel blocking properties. However, 1,9-dideoxy-forskolin was more potent than forskolin as a Ca2+ channel blocker. Furthermore, 1,9-dideoxy-forskolin was also more potent than forskolin as a nicotinic acetylcholine receptor and Ca2+ channel blocker and it was more potent as a nicotinic acetylcholine receptor blocker than Ca2+ channel blocker. The results showed powerful cAMP-independent effects of the diterpenes and suggest caution in interpretation of cAMP effects on chromaffin cells when its cellular levels are modified by forskolin. PMID- 9226414 TI - Neurospecificity of phyto-bufadienolides is not related to differences in Na+/K+ pump inhibition. AB - The aim of the present study was to investigate the effects of neuro- (cumulative) and cardiotoxic (non-cumulative) bufadienolides originating from plants (phyto-bufadienolides) on the Na+/K+ pump current (Ip) in cardiac (rat and guinea pig) and dorsal root ganglion cells (guinea pig), and on Ca2+ currents in cardiomyocytes (guinea pig). All bufadienolides tested (non-cumulative drugs: thesiuside, tyledoside C; lanceotoxin B and tyledoside F for the neurotoxic group) were potent blockers of Ip at concentrations in the micro- and submicromolar range. K0.5 values for Ip inhibition in dorsal root ganglion neurones were slightly lower compared to cardiomyocytes, but the order of potency was similar in both cell types. Both classes of bufadienolides were equipotent in suppressing Ip, generated by high- and low-affinity pump isoforms. Phenomena related to pump inhibition, as hypercontracture and increase in T-type Ca2+ current in cardiomyocytes, were influenced to the same extent. Therefore, from these results, neurospecificity of some bufadienolides could not be explained by differences in Na+/K+ pump affinity. PMID- 9226415 TI - Effects of thiocyanate and AMPA receptor ligands on (S)-5-fluorowillardiine, (S) AMPA and (R,S)-AMPA binding. AB - AMPA receptors can be labeled using the agonist radioligands [3H](R,S)-alpha amino-3-hydroxy-5-methylisoxazole-4-propionic acid ([3H](R,S)-AMPA), [3H](S)-AMPA or [3H](S)-5-fluorowillardiine. In the presence of KSCN, [3H](R,S)-AMPA and [3H](S)-AMPA bind to a single population of sites in rat brain membranes, whereas [3H](S)-5-fluorowillardiine binds with two affinity components. KSCN increased the affinity of the low affinity [3H](S)-5-fluorowillardiine component > 4-fold and increased the density of both components 1.5-1.7-fold, arguing against KSCN induced interconversion of low to high affinity states. KSCN, which promotes receptor desensitization, increased the potency of AMPA isomers, (S)-5 fluorowillardiine, quisqualate and cyclothiazide for inhibition of [3H](S)-5 fluorowillardiine binding suggesting that these ligands discriminate desensitized and nondesensitized receptors. In contrast, KSCN did not greatly affect the potency of glutamate, kainate, or competitive antagonists suggesting that these ligands do not discriminate desensitized and nondesensitized receptors. In the presence of KSCN, the rank order potency for agonists and antagonists was similar or identical in all assays indicating that the three radioligands bind identical glutamate recognition sites, a conclusion supported by their identical total receptor density. However, AMPA isomers displayed 6-10-fold higher potency for displacement of [3H](S)- or (R,S)-AMPA relative to [3H](S)-5-fluorowillardiine binding. This finding, coupled with the marked two component binding by [3H](S)-5 fluorowillardiine but not [3H](S)- or (R,S)-AMPA, suggests qualitative differences between the interaction of these ligands with the agonist recognition site. PMID- 9226416 TI - Unsaturated phosphinic analogues of gamma-aminobutyric acid as GABA(C) receptor antagonists. AB - The phosphinic and methylphosphinic analogues of gamma-aminobutyric acid (GABA) are potent GABA(C) receptor antagonists but are even more potent as GABA(B) receptor agonists. Conformationally restricted unsaturated phosphinic and methylphosphinic analogues of GABA and some potent GABA(B) receptor phosphonoamino acid antagonists were tested on GABA(C) receptors in Xenopus oocytes expressing human retinal rho1 mRNA. 3-Aminopropyl-n-butyl-phosphinic acid (CGP36742), an orally active GABA(B) receptor antagonist, was found to be a moderately potent GABA(C) receptor antagonist (IC50 = 62 microM). The unsaturated methylphosphinic and phosphinic analogues of GABA were competitive antagonists of the GABA(C) receptors, the order of potency being [(E)-3-aminopropen-1 yl]methylphosphinic acid (CGP44530, IC50 = 5.53 microM) > [(E)-3-aminopropen-1 yl]phosphinic acid (CGP38593, IC50 = 7.68 microM) > [(Z)-3-aminopropen-1 yl]methylphosphinic acid (CGP70523, IC50 = 38.94 microM) > [(Z)-3-aminopropen-1 yl]phosphinic acid (CGP70522, IC50 > 100 microM). This order of potency differs from that reported for these compounds as GABA(B) receptor agonists, where the phosphinic acids are more potent than the corresponding methylphosphinic acids. PMID- 9226418 TI - Corticotropin-releasing factor-like peptides increase cytosolic [Ca2+] in human epidermoid A-431 cells. AB - This study investigated whether sauvagine and urotensin I change [Ca2+]i in human epidermoid A-431 cells and whether these changes are correlated with their anti edema properties in vivo. A-431 cells were used because they possess the corticotropin-releasing factor (CRF) receptor 2. Treatment with either sauvagine or urotensin I led to an immediate increase in [Ca2+]i, the magnitude of which depended on the concentration of the drug. Sauvagine was more effective than urotensin I, with a median effective concentration (EC50) of 1.4 +/- 0.2 fM, compared to an EC50 of 66 +/- 7 fM for urotensin I. Both were more effective at stimulating increases in [Ca2+]i than CRF (EC50 of 6.8 +/- 0.1 pM). There was a correlation between the EC50 for increasing [Ca2+]i and the median effective dose (ED50) for inhibiting edema induced by heating rat paw (r = 0.99). Removal of extracellular Ca2+ or incubation with La3+ eliminated the increase in [Ca2+]i stimulated by either peptide. Pretreatment with a CRF receptor antagonist reduced the increase in [Ca2+]i by these peptides. This occurred in an antagonist concentration-dependent manner, with median inhibitory concentrations (IC50) of 1.99 +/- 0.04 nM and 0.85 +/- 0.04 nM, respectively. Both pertussis toxin (an inhibitor of G proteins) and U-73122 (an inhibitor for inositol trisphosphate (InsP3) production) partially inhibited the increases. InsP3 was measured to determine whether these peptides mobilized Ca2+ from an InsP3-sensitive store. Both sauvagine and urotensin I increased InsP3. The InsP3 increases were inhibited by U-73 122 and CRF antagonist, but not by removal of external Ca2+. Both peptides elevated protein tyrosine phosphorylation. In summary, these peptides increase [Ca2+]i as a result of Ca2+ influx via CRF receptor-operated Ca2+ channels coupled to pertussis toxin-sensitive G proteins and a Ca2+ mobilization from InsP3-sensitive Ca2+ pools. Their in vivo effectiveness at inhibiting edema is related to their respective capacities to stimulate elevations of [Ca2+]i, supporting a role for intracellular Ca2+ in this process. PMID- 9226417 TI - Slow desensitization of the human P2Y6 receptor. AB - The P2Y6 receptor is a recently cloned P2 receptor which displays a high sensitivity for diphosphonucleotides. In 1321N1 astrocytoma cells stably expressing this receptor, UDP induced a slow and sustained accumulation of inositol trisphosphate via a pertussis toxin-insensitive G-protein: the maximal level was only reached after 15 min and a significant response was maintained for at least 3 h. A full second response to UDP was obtained after the first 45-min stimulation, but was lost after 165 min. This slow and sustained time-course and the lack of desensitization was reproduced with ADP. UTP was unable to restimulate the P2Y4 receptor, another recently cloned P2 receptor with a preference for UTP, after the first 5-min stimulation. The P2Y4 receptor is thus rapidly desensitized whereas desensitization of the P2Y6 receptor is delayed. The rank order of potency of various diphosphonucleotides at the P2Y6 receptor was: UDP > TDP > IDP > GDP > ADP >> CDP. The activity of three non-specific antagonists of P2 receptors was characterized by the following rank order of potency: reactive blue 2 > pyridoxal-phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) > suramin. In conclusion, the most impressive features of the human P2Y6 receptor revealed by this study are the slow and sustained time-course of its activation and its high resistance to desensitization. PMID- 9226419 TI - Indomethacin and piroxicam inhibit Na+-adenosine transport in rat renal brush border membranes. AB - The effects of the cyclooxygenase inhibitors, indomethacin and piroxicam, were evaluated on Na+-dependent [3H]adenosine transport in rat renal brush-border membranes of the outer renal cortex of the rat. Adenosine co-transport (1-10 microM) was estimated in the presence of 0.001-10 microM indomethacin and piroxicam. Both drugs inhibited the Na+-dependent transport in a dose-dependent manner with IC50 of 3.5 microM and 0.1 microM, respectively. The Na+-independent transport was not modified. Preincubations carried out on the vesicles with 10-50 microM arachidonic acid increased transport in a dose-dependent manner up to 1.7 times. Whereas 50 pM to 5 microM prostaglandin E2 in the presence of indomethacin did not change carrier activity, 5 microM prostaglandin E2 increased the Na+ dependent transport 1.5 times. Other prostanoid synthesis pathways were investigated with 10 microM nordihydroguaiaretic acid (lipoxygenase inhibitor), and 17-octadecynoic acid and clotrimazole (leukotriene and cytochrome P450 inhibitors). Our results demonstrated that the Na+-dependent adenosine transport in brush-border membranes was inhibited by indomethacin and piroxicam, suggesting that cyclooxygenase activity might modulate this co-transport. PMID- 9226420 TI - Decreased agonist sensitivity of human GABA(A) receptors by an amino acid variant, isoleucine to valine, in the alpha1 subunit. AB - Recombinant human GABA(A) receptors were investigated in vitro by coexpression of cDNAs coding for alpha1, beta2, and gamma2 subunits in the baculovirus/Sf-9 insect cell system. We report that a single amino acid exchange (isoleucine 121 to valine 121) in the N-terminal, extracellular part of the alpha1 subunit induces a marked decrease in agonist GABA(A) receptor ligand sensitivity. The potency of muscimol and GABA to inhibit the binding of the GABA(A) receptor antagonist [3H]SR 95531 (2-(3-carboxypropyl)-3-amino-6-(4 methoxyphenyl)pyridazinium bromide) was higher in receptor complexes of alpha1(ile 121) beta2gamma2 than in those of alpha1(val 121) beta2gamma2 (IC50 values were 32-fold and 26-fold lower for muscimol and GABA, respectively). The apparent affinity of the GABA(A) receptor antagonist bicuculline methiodide to inhibit the binding of [3H]SR 95531 did not differ between the two receptor complex variants. Electrophysiological measurements of GABA induced whole-cell Cl currents showed a ten-fold decrease in the GABA(A) receptor sensitivity of alpha1 (val 121) beta2gamma2 as compared to alpha1(ile 121) beta2gamma2 receptor complexes. Thus, a relatively small change in the primary structure of the alpha1 subunit leads to a decrease selective for GABA(A) receptor sensitivity to agonist ligands, since no changes were observed in a GABA(A) receptor antagonist affinity and benzodiazepine receptor binding. PMID- 9226421 TI - Anethole dithiolethione prevents oxidative damage in glutathione-depleted astrocytes. AB - Astrocytes protect neurons against reactive oxygen species such as hydrogen peroxide, a capacity which reportedly is abolished following loss of the antioxidant glutathione. Anethole dithiolethione, a sulfur-containing compound which is used in humans, is known to increase cellular glutathione levels and thought thereby to protect against oxidative damage. In the present study we found that anethole dithiolethione increased the glutathione content of cultured rat striatal astrocytes. This effect was abolished by coincubation with the glutathione synthesis inhibitor buthionine sulfoximine. Nevertheless, in the presence of buthionine sulfoximine, despite the lack of an increase in the lowered glutathione level, anethole dithiolethione fully protected the astrocytes against the enhanced toxicity of hydrogen peroxide. Thus, apparently other mechanisms than stimulation of glutathione synthesis are involved in the compound's protective action in astrocytes. Considering the occurrence of lowered glutathione levels in neurodegenerative syndromes, we conclude that further evaluation of the therapeutic potential of anethole dithiolethione is warranted. PMID- 9226422 TI - Occlusal imbalance and temporomandibular disorders in the elderly. AB - The prevalence of the anamnestic symptoms and clinical signs of temporomandibular disorders (TMD) in 76-, 81-, and 86-year-old subjects was studied, on the basis of Helkimo's anamnestic (Ai) and clinical (Di) dysfunction indexes. Occlusal status was recorded by means of the Eichner index: class A has a maximum of four supporting zones (minimum of one tooth contact between the antagonist jaws in the premolar and molar region on each side), class B has one to three supporting zones or tooth contact in the frontal area only, and class C has no supporting zones. The Eichner index was recorded with two kinds of variations: supporting zones with and without removable prostheses. In the population studied 8% were classified as Eichner class A, 22% as class B, and 70% as class C. Including occlusal supporting zones of the removable dentures, the percentages were 75% in Eichner class A, 21% in class B and 4% in class C. When the groups with and without removable prostheses were compared, no differences were found in the Ai or Di. In conclusion, among the elderly population the severity of TMD does not depend on the supporting zones of the dentition alone, and removable prostheses do not relieve the problem. PMID- 9226424 TI - Dental management of Alzheimer patients. A predictive test of dental cooperation in individualized treatment planning. AB - The aim of the study was to investigate a possible relationship in Alzheimer patients between the stage of dementia, cognitive and functional capacity, and behavior as a dental patient. A special index for assessing behavior in the dental setting was used. The 40 participants were inmates of a nursing home and fulfilled the criteria for Alzheimer's disease in accordance with DSM-III-R. A deficiency in the dental behavior index (DBI) of 50% or less did not correlate with cognitive, functional, or graphic capacity. These subjects were generally aware of earlier regular dental treatment and behaved as if dental visits were familiar to them, although their cognitive, functional, and graphic capacities were more impaired than disclosed by the dental behavior index. A deficiency in the dental behavior index of 50% or more was more correlated with the other capacity assessments. Finding a proper treatment level for an Alzheimer patient is a delicate task in which it is essential to balance awareness of various aspects of impairment and realistic anticipation of benefit. The dental behavior index can be an appropriate instrument in this complex process. PMID- 9226423 TI - Subjective aspects of patients with traumatized teeth. A 15-year follow-up study. AB - The aim of this study was to obtain detailed information about adults who suffered trauma to the teeth as children. A total of 102 patients took part. The patients answered a questionnaire and were interviewed before the oral examination. Thirty-nine per cent of the patients reported dissatisfaction either with the color and/or anatomic form of the traumatized teeth or reconstruction. Most of the individuals did not remember having received any information about prognosis for the traumatized teeth. Twenty-one per cent of the patients remembered pain during treatment, and 25% remembered only the behavior and attitudes of the dental team. It can be concluded that all dental treatment in children with traumatized teeth must be carried out as painlessly as possible, and the dental team should minimize discomfort during the treatment. Good knowledge about traumatology and management can reduce stress and anxiety for both the patient and the dental team. PMID- 9226426 TI - Visible-light curing units: correlation between depth of cure and distance between exit window and resin surface. AB - Study 1) The depth of proximal cavities was measured on previously taken bitewing radiographs. Study 2) By means of a scrape test, the relationship between depth of cure and irradiation distance was examined with four different curing units and two different restorative resins. Study 3) The microhardness of one of the polymerized resins was measured 0.5 mm below the free surface of the filling and then at 1.0 mm, 1.5 mm, 2.0 mm, and so forth until the resin became so soft that no hardness could be recorded. The hardness at each 0.5-mm level was recorded in relation to irradiation distance. 1) The cavity depth was most often 4-5 mm in lower premolars, 5 6 mm in upper premolars and lower molars, and 5-7 mm in upper molars. In the latter teeth, 15% of the cavities were > or =8 mm deep. 2 and 3) The depth of cure decreased moderately and in a linear manner with increasing irradiation distance. An irradiation distance of 12 mm reduced the depth of well cured resin only by about 1 mm as compared with close contact between exit window and surface of resin. PMID- 9226425 TI - Quantitative assessment of vertical heights of maxillary and mandibular bones in panoramic radiographs of elderly dentate and edentulous subjects. AB - The clinical applicability of vertical measurements of the mandible and maxilla in panoramic radiographs was studied by assessing the variety of vertical heights among 91 elderly dentate subjects. Measurements in each jaw and calculations of a maxillary ratio were made at five sites. Variations in measurements of the dentate subjects were small: 9-11% for vertical measurements in the mandible, 6 11% for vertical measurements in the maxilla, and 8-10% for the maxillary ratios. These findings suggest that it is possible quantitatively to assess heights of the mandibular and maxillary bones in panoramic radiographs. Reductions in the edentulous jaws were assessed by comparing the heights of jaws of elderly dentate subjects with those measured in 177 elderly edentulous subjects. Significant differences in heights of the mandibular body and maxilla were found between the dentate and the edentulous (P < 0.001). Edentulous women had greater values for percentage reduction in the mandibles than did the men (P < 0.01; P < 0.001 in various locations). PMID- 9226427 TI - Bone volume in human temporomandibular autopsy joints with and without erosive changes. AB - The aim was to compare the trabecular bone volume (TBV) and the total bone volume (TOBV) of human temporomandibular joints (TMJ) with erosive changes with those of joints without erosive changes. We examined 35 TMJ autopsy specimens from 19 individuals aged 66 88 years. Sagittal sections of the joints were analyzed microscopically for erosive hard-tissue changes. The TBV and the TOBV of the sections were assessed with the aid of computerized image analysis. A significant increase in trabecular and total bone volume was found in condyles with erosive changes (TBV = 21%, TOBV = 54%) as compared with condyles without erosive changes (TBV = 15%, TOBV = 40%). The trabecular bone volume of the temporal component was also increased (TBV = 24%) in joints with erosive changes in the condyle as compared with joints with unaffected condyles (TBV = 16%). The findings indicate that the relative bone mass may play a role in the development of erosive changes in the TMJ. PMID- 9226428 TI - Direct evidence concerning the 'major role' of fluoride dentifrices in the caries decline. A 6-year analytical cohort study. AB - The role of fluoride (F) dentifrices in caries decline was investigated by assessing the effect of variation in their use on caries scores among teenagers. The material comprised 211 subjects aged about 11 years at base line and 18 years at the last examination. Pairs of posterior bitewing radiographs were assessed by one examiner. Information concerning dental health behavior was collected by questionnaire and about treatment received from dental records. A reversal of the traditional DFS gender difference occurred during teenage years. Multivariate regression analyses showed an inverse relationship between variation in F dentifrice use and current decay (D1S) at age 18 years (P < 0.04) and with caries incidence per year (D1FS) during the whole observation period (P < 0.02). Total explained variance in 6-year deltaD1FS scores was 29.8%, of which variation in toothbrushing behavior contributed 1.8 percentage points. While confirming the multifactorial nature of dental caries, these results also provided quantitative evidence for the role of variation in F dentifrice use in caries incidence and decline. PMID- 9226429 TI - Management of chronic orofacial pain: attitudes among patients and dentists in a Swedish county. AB - The purpose was to survey attitudes towards management of chronic orofacial pain (COP). Questionnaires were mailed to 30 randomized dentists and to 30 consecutive COP patients, examined 16 months earlier by a pain group of dental specialists. Fifty-seven per cent of the patients reported that their pain was the same as or worse than before and was disturbing. Few were dissatisfied with the examinations. Fifty-nine per cent thought that the consultations had been good. The surveyed dentists judged the most common causes of COP to be neurogenic and psychogenic in origin; they were overwhelmingly positive to the idea of a pain group (93%) and could consider referring patients (97%). Pain-inducing local diseases occurred but were not dominant among these COP patients. We concluded that management of COP in a pain group appears to be meaningful, as reflected by the respondents' attitudes but would gain by a closer collaboration with medical expertise. PMID- 9226430 TI - A scanning electron microscopy study of disturbances in the developing rat molar induced by cyclophosphamide. AB - Scanning electron microscopy was used to study the effect of cyclophosphamide (Cy) on molar development in 18 Sprague-Dawley rats from 15 to 48 days of age after birth. Doses of 30 mg/kg body weight of Cy dissolved in 1 ml 0.9% NaCl were given to the rats at 10 and 13 days of age. Eighteen control rats had injections of 1 ml 0.9% NaCl at the same ages. The most obvious changes in the experimental teeth were found in the developing roots of the first and second molars and in both the crown and roots of the third molar. The roots of the first and second molars were short and showed apical closure in the experimental rats. In addition to the disturbances in crown and root formation, the third molars were also significantly reduced in total size as compared with the third molars in the control rats. PMID- 9226431 TI - Dental health and dental treatment needs among recruits of the Finnish Defence Forces, 1919-91. AB - The first two surveys of the dental health of young Finnish men were conducted in 1919 and 1965. The objective of four subsequent surveys (1976, 1981, 1986, and 1991) was to collect both interview and clinical examination data for the monitoring of changes in the oral health status of the recruits. A significant reduction in self-reported toothache, gingival bleeding, and number of decayed teeth was observed from 1976 to 1991. At examination, the numbers of decayed teeth, teeth indicated for extraction, teeth in need of fillings, and missing teeth decreased substantially, as did the teeth with visible plaque, subgingival calculus, and teeth with 4-mm or deeper periodontal pockets. This comprehensive series of successive cross-sectional oral health surveys clearly shows that since 1976 a significant decrease in oral disease and treatment needs has taken place among the Finnish population of young men. PMID- 9226432 TI - The effect of zinc-containing chewing gum on volatile sulfur-containing compounds in the oral cavity. AB - Volatile sulfur-containing compounds (VSC) are known to constitute the major component of halitosis. Aqueous solutions of zinc salts have been shown to reduce the levels of VSC produced orally. The aim of the present study was to examine whether zinc could be made available in the oral cavity and inhibit VSC production when delivered by a chewing gum. VSC measurements were carried out on the 'morning breath' of 11 test subjects and re-examined after the use of test solutions containing 0.02% zinc chloride, 0.2% chlorhexidine, or water or the use of chewing gums containing 2 mg, 0.5 mg, or 0 mg zinc acetate. The results showed that similar amounts of zinc in mouthrinses or chewing gum had the same effect, with a reduction of the oral VSC of 45%. Chewing gum thus seems to be a viable alternative for delivering zinc to reduce VSC levels in the oral cavity. PMID- 9226433 TI - Evolution of B2 repeats: the muroid explosion. AB - B2 repeats are a group of short interspersed elements (SINEs) specific for rodent genomes. Copy numbers were determined for different rodent genera. All the Muroid (rat, mouse, deer mouse, hamster, gerbil) rodent genomes analyzed exhibited 80,000-100,000 copies per haploid genome, whereas the squirrel genome contains only 2,500 copies, and fewer than 100 (if any) copies were observed for the Hystricognath rodents (guinea pig and nutria). These findings demonstrate that there was an 'explosion' of amplification of B2 elements within muroid rodents. The similar copy number of B2 elements within the different muroid species could be explained by formation of a high proportion of the B2 elements prior to the divergence of the different muroid species. However, the 3'-end of the B2 sequence is unique between murid and cricetid rodents suggesting that the majority of elements amplified after the divergence of these species. Also consistent with recent amplification of these elements in parallel within the muroid genomes is the finding that within mouse and rat there are distinct subfamilies of B2 repeats. The pattern of consistent parallel amplification of B2 elements in muroid species contrasts with the sporadic nature of ID repeat amplification in the same genomes. The consensus of the young mouse subfamily of elements corresponds to the B2 RNA that is preferentially transcribed in embryonic, tumor, and normal liver cells. The subfamily is young based on both its low divergence from the subfamily consensus sequence and the finding that the most recent B2 element insertions in the mouse genome are members of this subfamily. PMID- 9226435 TI - Genetical analysis of visual system disorganizer (vid), a new gene involved in normal development of eye and optic lobe of the brain in Drosophila melanogaster. AB - A neuroanatomical screening of a collection of P-element mutagenized flies has been carried out with the aim of finding new mutants affecting the optic lobe of the adult brain in Drosophila melanogaster. We have identified a new gene that is involved in the development of the adult axon array in the optic ganglia and in the ommatidia assembly. We have named this locus visual system disorganizer (vid). Reversional mutagenesis demonstrated that the vid mutant was the result of a P-element insertion in the Drosophila genome and allowed us to generate independent alleles, some of which resulted in semilethality, like the vid original mutant, while the others were completely lethal. A genetic somatic mosaic analysis indicated that the vid gene is required in the eye for its normal development by inductive effects. This analysis also suggests an inductive effect of the vid gene on the distal portion of the optic lobe, particularly the lamina and the first optic chiasma. Moreover, the absence of mutant phenotype in the proximal region of the optic ganglia, including the medulla, the second optic chiasma, and the lobula complex underlying mosaic eyes, is suggestive of an autonomously acting mechanism of the vid gene in the optic lobe. The complete or partial lethality generated by different mutations at the vid locus suggests that this gene's role may not be limited to the visual system, but may also affect a vital function during Drosophila development. PMID- 9226434 TI - The Hermes element from Musca domestica can transpose in four families of cyclorrhaphan flies. AB - Transgenic insect technology will provide opportunities to explore the basic biology of a broad range of insect species in ways that will prove insightful and important. It is also a technology that will provide opportunities to manipulate the genotypes of insects of practical significance to the health and welfare of humans. The Hermes transposable element from the housefly, Musca domestica, is a short inverted repeat-type element related to hobo from Drosophila melanogaster, Ac from Zea mays, and Tam3 from Antirrhinum majus. It has potential to become a versatile and efficient broad host-range insect transformation vector. The ability of Hermes to transpose when introduced into five species of diptera from four divergent families was tested using an in vivo, interplasmid transpositional recombination assay. Hermes was capable of transposing in all species tested, demonstrating that Hermes has a broad host-range. In addition, the rates of transposition were sufficiently high in all species tested to suggest that Hermes will be an efficient gene transfer vector in a wide range of insect species. The Hermes element also revealed a pattern of integration into the target substrate that permitted factors determining integration site selection to be identified. Primary nucleotide sequence of the integration site played a role as did proximity to preferred integration sites and the nucleosomal organization of the target. PMID- 9226436 TI - Bootstrap tests for specific hypotheses at single locus inbreeding coefficients. AB - Deviations of genotype distribution from Hardy-Weinberg expectations within a (sub)population can give valuable insight into the population structure, and can be quantified by means of F(is) values. Specific biological and/or genetical hypotheses regarding F(is) require particular statistical procedures to be able to perform the test with high power. The bootstrap offers a convenient way to test against a broad range of alternative hypotheses. It enables: a) comparison of an observed F(is) with any expected value between -1 and 1, and b) comparison of two or more observed F(is) values. However, it fails under numerous situations, and great caution should be taken before applying the bootstrap to estimate confidence intervals of F(is). We discuss under which conditions the bootstrap gives reliable results. PMID- 9226437 TI - Non-linear selection response in Drosophila: a strategy for testing the rare alleles model of quantitative genetic variability. AB - Quantitative genetic theory predicts that variation due to rare alleles at many loci will generate a transient acceleration in the response to directional selection. We have tested this prediction by constructing experimental lines of Drosophila melanogaster that carry positively selected ethanol resistance alleles at low frequencies, and then subjecting the lines to directional selection for ethanol resistance. Approximately 468,000 files were subjected to artificial selection over 30 generations. The predicted non-linear selection responses were observed in all experimental lines and replicates, on three genetic backgrounds. In contrast, un-selected controls and lines carrying random alleles at low frequencies on the same genetic backgrounds exhibited linear selection responses. These results demonstrate that non-linearities due to rare alleles are detectable and repeatable, provided that experiments are done on a sufficiently large scale. The results suggest that it may be possible to test for rare-alleles as a component of naturally occurring genetic variation by careful examination of selection response curves. PMID- 9226438 TI - Regulation of cone cell formation by Canoe and Ras in the developing Drosophila eye. AB - Cone cells are lens-secreting cells in ommatidia, the unit eyes that compose the compound eye of Drosophila. Each ommatidium contains four cone cells derived from precursor cells of the R7 equivalence group which express the gene sevenless (sev). When a constitutively active form of Ras1 (Ras1V12) is expressed in the R7 equivalence group cells using the sev promoter (sev-Ras1V12), additional cone cells are formed in the ommatidium. Expression of Ras1N17, a dominant negative form of Ras1, results in the formation of 1-3 fewer cone cells than normal in the ommatidium. The effects of Ras1 variants on cone cell formation are modulated by changing the gene dosage at the canoe (cno) locus, which encodes a cytoplasmic protein with Ras-binding activity. An increase or decrease in gene dosage potentiates the sev-Ras1v12 action, leading to marked induction of cone cells. A decrease in cno+ activity also enhances the sev-Ras1N17 action, resulting in a further decrease in the number of cone cells contained in the ommatidium. In the absence of expression of sev-Ras1V12 or sev-Ras1N17, an overdose of wild-type cno (cno+) promotes cone cell formation while a significant reduction in cno+ activity results in the formation of 1-3 fewer cone cells than normal in the ommatidium. We propose that there are two signaling pathways in cone cell development, one for its promotion and the other for its repression, and Cno functions as a negative regulator for both pathways. We also postulate that Cno predominantly acts on a prevailing pathway in a given developmental context, thereby resulting in either an increase or a decrease in the number of cone cells per ommatidium. The extra cone cells resulting from the interplay of Ras1v12 and Cno are generated from a pool of undifferentiated cells that are normally fated to develop into pigment cells or undergo apoptosis. PMID- 9226439 TI - Induction of identified mesodermal cells by CNS midline progenitors in Drosophila. AB - The Drosophila ventral midline cells generate a discrete set of CNS lineages, required for proper patterning of the ventral ectoderm. Here we provide the first evidence that the CNS midline cells also exert inductive effects on the mesoderm. Mesodermal progenitors adjacent to the midline progenitor cells give rise to ventral somatic mucles and a pair of unique cells that come to lie dorsomedially on top of the ventral nerve cord, the so-called DM cells. Cell ablation as well as cell transplantation experiments indicate that formation of the DM cells is induced by midline progenitors in the early embryo. These results are corroborated by genetic analyses. Mutant single minded embryos lack the CNS midline as well as the DM cells. Embryos mutant for any of the spitz group genes, which primarily express defects in the midline glial cell lineages, show reduced formation of the DM cells. Conversely, directed overexpression of secreted SPITZ by some or all CNS midline cells leads to the formation of additional DM cells. Furthermore we show that DM cell development does not depend on the absolute concentration of a local inductor but appears to require a graded source of an inducing signal. Thus, the Drosophila CNS midline cells play a central inductive role in patterning the mesoderm as well as the underlying ectoderm. PMID- 9226440 TI - The PDGF alpha receptor is required for neural crest cell development and for normal patterning of the somites. AB - Platelet-derived growth factors (PDGFs) have been implicated in the control of cell proliferation, survival and migration. Patch mutant mice harbor a deletion including the PDGF alpha receptor gene and exhibit defects of neural crest origin which affect pigmentation in heterozygotes and cranial bones in homozygotes. To verify the role of the PDGF alphaR gene during development, mice carrying a targeted null mutation were generated. No pigmentation phenotype was observed in heterozygotes. Homozygotes die during embryonic development and exhibit incomplete cephalic closure similar to that observed in a subset of Patch mutants. In addition, increased apoptosis was observed on pathways followed by migrating neural crest cells. However, alterations in mutant vertebrae, ribs and sternum were also observed, which appear to stem from a deficiency in myotome formation. These results indicate that PDGFs may exert their functions during early embryogenesis by affecting cell survival and patterning. PMID- 9226441 TI - Inhibins and activins regulate mammary epithelial cell differentiation through mesenchymal-epithelial interactions. AB - Inhibins and activins are members of the transforming growth factor beta (TGFbeta) family. Female mice in which both alleles encoding the inhibin betaB subunit have been deleted are unable to nurse their pups. We have now identified a cause of lactation failure in these mice. Ductal elongation and alveolar morphogenesis are retarded. During puberty and pregnancy, ductal outgrowth and alveolar development are limited and morphologically abnormal endbuds persist in the glands of postpartum females. The alveolar lumina fail to expand at parturition due to the absence of secreted milk. Transplantation experiments have been performed to determine whether the absence of systemic- or mammary-derived betaB subunits are the cause for the incomplete and aberrant development. While transplanted intact glands from wild-type mice grew normally in betaB-deficient hosts, betaB-deficient glands remained underdeveloped in wild-type hosts. However, betaB-deficient epithelium developed normally when transplanted into the fat pad of wild-type hosts. This demonstrates that ductal elongation and epithelial cell differentiation during puberty and pregnancy require activin/inhibin signalling from the stroma. The results further show that distinct, though related, activins and inhibins perform unique functions and are not able to compensate for the absence of activin B and AB and inhibin B in the process of mammogenesis. The betaB-deficient mice provide the first genetic evidence for stromal signalling in the adult mammary gland in vivo. PMID- 9226443 TI - cudA: a Dictyostelium gene with pleiotropic effects on cellular differentiation and slug behaviour. AB - The Dictyostelium cudA gene encodes a nucleoplasmic protein that is essential for normal culmination. There are no functionally characterised homologues in other organisms but there is a related gene of unknown function in Entamoeba histolytica. The cudA gene is expressed by the prestalk cells that constitute the slug tip (the pstA cells), it is not detectably expressed in the band of prestalk cells that lies behind the tip (the pstO cells) but it is expressed in the prespore cells. This unusual pattern of expression suggests a role on both the stalk and spore pathways of differentiation and cudA- mutant cells are indeed defective in both stalk and spore formation. Furthermore, the slugs formed by cudA- cells continue to migrate under environmental conditions where normal slugs culminate immediately. This aspect of their behaviour can be reversed when the cudA gene is selectively expressed in the pstA cells. This shows that processes occurring in the pstA cells regulate entry into culmination. PMID- 9226442 TI - Inductive interactions direct early regionalization of the mouse forebrain. AB - The cellular and molecular mechanisms that regulate regional specification of the forebrain are largely unknown. We studied the expression of transcription factors in neural plate explants to identify tissues, and the molecules produced by these tissues, that regulate medial-lateral and local patterning of the prosencephalic neural plate. Molecular properties of the medial neural plate are regulated by the prechordal plate perhaps through the action of Sonic Hedgehog. By contrast, gene expression in the lateral neural plate is regulated by non-neural ectoderm and bone morphogenetic proteins. This suggests that the forebrain employs the same medial-lateral (ventral-dorsal) patterning mechanisms present in the rest of the central nervous system. We have also found that the anterior neural ridge regulates patterning of the anterior neural plate, perhaps through a mechanism that is distinct from those that regulate general medial-lateral patterning. The anterior neural ridge is essential for expression of BF1, a gene encoding a transcription factor required for regionalization and growth of the telencephalic and optic vesicles. In addition, the anterior neural ridge expresses Fgf8, and recombinant FGF8 protein is capable of inducing BF1, suggesting that FGF8 regulates the development of anterolateral neural plate derivatives. Furthermore, we provide evidence that the neural plate is subdivided into distinct anterior posterior domains that have different responses to the inductive signals from the prechordal plate, Sonic Hedgehog, the anterior neural ridge and FGF8. In sum, these results suggest that regionalization of the forebrain primordia is established by several distinct patterning mechanisms: (1) anterior-posterior patterning creates transverse zones with differential competence within the neural plate, (2) patterning along the medial-lateral axis generates longitudinally aligned domains and (3) local inductive interactions, such as a signal(s) from the anterior neural ridge, further define the regional organization. PMID- 9226444 TI - Dorsal hindbrain ablation results in rerouting of neural crest migration and changes in gene expression, but normal hyoid development. AB - Our previous studies have shown that hindbrain neural tube cells can regulate to form neural crest cells for a limited time after neural fold removal (Scherson, T., Serbedzija, G., Fraser, S. E. and Bronner-Fraser, M. (1993). Development 188, 1049-1061; Sechrist, J., Nieto, M. A., Zamanian, R. T. and Bronner-Fraser, M. (1995). Development 121, 4103-4115). In the present study, we ablated the dorsal hindbrain at later stages to examine possible alterations in migratory behavior and/or gene expression in neural crest populations rostral and caudal to the operated region. The results were compared with those obtained by misdirecting neural crest cells via rhombomere rotation. Following surgical ablation of dorsal r5 and r6 prior to the 10 somite stage, r4 neural crest cells migrate along normal pathways toward the second branchial arch. Similarly, r7 neural crest cells migrate primarily to the fourth branchial arch. When analogous ablations are performed at the 10-12 somite stage, however, a marked increase in the numbers of DiI/Hoxa-3-positive cells from r7 are observed within the third branchial arch. In addition, some DiI-labeled r4 cells migrate into the depleted hindbrain region and the third branchial arch. During their migration, a subset of these r4 cells up-regulate Hoxa-3, a transcript they do not normally express. Krox20 transcript levels were augmented after ablation in a population of neural crest cells migrating from r4, caudal r3 and rostral r3. Long-term survivors of bilateral ablations possess normal neural crest-derived cartilage of the hyoid complex, suggesting that misrouted r4 and r7 cells contribute to cranial derivatives appropriate for their new location. In contrast, misdirecting of the neural crest by rostrocaudal rotation of r4 through r6 results in a reduction of Hoxa-3 expression in the third branchial arch and corresponding deficits in third arch-derived structures of the hyoid apparatus. These results demonstrate that neural crest/tube progenitors in the hindbrain can compensate by altering migratory trajectories and patterns of gene expression when the adjacent neural crest is removed, but fail to compensate appropriately when the existing neural crest is misrouted by neural tube rotation. PMID- 9226445 TI - DPP controls tracheal cell migration along the dorsoventral body axis of the Drosophila embryo. AB - We report that DPP signaling is required for directed tracheal cell migration during Drosophila embryogenesis. The failure of tracheal cells to receive the DPP signal from adjacent dorsal and ventral cells results in the absence of dorsal and ventral migrations. Ectopic DPP signaling can reprogram cells in the center of the placode to adopt a dorsoventral migration behavior. The effects observed in response to ectopic DPP signaling are also observed upon the tracheal-specific expression of a constitutive active DPP type I receptor (TKV(Q253D)), indicating that the DPP signal is received and transmitted in tracheal cells to control their migration behavior. DPP signaling determines localized gene expression patterns in the developing tracheal placode, and is also required for the dorsal expression of the recently identified BRANCHLESS (BNL) guidance molecule, the ligand of the BREATHLESS (BTL) receptor. Thus, DPP plays a dual role during tracheal cell migration. It is required to control the dorsal expression of the BNL ligand; in addition, the DPP signal recruits groups of dorsal and ventral tracheal cells and programs them to migrate in dorsal and ventral directions. PMID- 9226446 TI - The role in neural patterning of translation initiation factor eIF4AII; induction of neural fold genes. AB - Expression of the RNA-helicase translation initiation factor, eIF4AII, in animal cap explants of Xenopus specifically upregulates genes expressed early in the neural plate border such as Xsna, Xslu, Pax-3 and XANF and also the cement gland marker XCG-1. eIF4AII is expressed specifically in the prospective neurectoderm from stage 11.5 and appears to have a significant role in mediating early patterning of the neurectoderm. It is induced by all known neural inducing regimes including secreted factors such as noggin, follistatin and chordin, transcription factors such as XlPou-2 and constructs that overcome repression of neural induction (tBMP-4R, lim-m3 and Xbra delta 304). It is also upregulated when neurulization occurs in embryonic ectoderm that has been disaggregated and reaggregated. While high amounts of injected mRNA of the neural inducers noggin, tBMP-4R and Xlpou-2 downregulate Xslu and upregulate the neural plate NCAM, smaller amounts of these mRNAs activate expression of eIF4AII and Xslu and suppress expression of epidermal keratin in animal cap assays. Ectopic expression of eIF4AII mRNA also upregulates transcription of the PKC alpha and beta genes. The sensitivity of the upregulation of neurectodermal markers to GF109203X indicates that the activity of a calcium activated protein kinase C (PKC) is also required. Furthermore ectopic expression of mouse eIF4AII mRNA upregulates the endogenous eIF4AII gene by a process that requires the activity of PKC. The effects of eIF4AII appear to be direct as conditional expression of eIF4AII in animal cap explants at the equivalent of stage 11.5 induces the endogenous eIF4AII and neural fold genes within 40 minutes. Expression of eIF4AII and activation of PKC sensitizes the embryonic ectoderm to the neuralising effect of noggin. We suggest that in developing embryos the neuralizing signal emanating from the organiser at first induces eIF4AII and the prospective neural crest in an arc low on the dorsal aspect of the embryo. As the neuralizing signal increases in intensity close to the organizer region, the tissue becomes committed to a neural plate phenotype. Expression of Xash-3A may suppress further expression of neural plate border genes within the prospective neural plate thereby subdividing the neurectoderm into two distinct regions. PMID- 9226447 TI - Cellularization in locust embryos occurs before blastoderm formation. AB - In Drosophila intracellular gradients establish the pattern of segmentation by controlling gene expression during a critical syncytial stage, prior to cellularization. To investigate whether a similar mechanism may be exploited by other insects, we examined the timing of cellularization with respect to blastoderm formation in an insect with extreme short-germ development, the African desert locust, Schistocerca gregaria. Using light and electron microscopic techniques, we show that the islands of cytoplasm surrounding cleavage nuclei are largely isolated from their neighbours, allowing cleavage to proceed asynchronously. Within a short time of their arrival at the surface and prior to blastoderm formation, nuclei become surrounded by complete cell membranes that block the free uptake of dye (10,000 kDa) from the yolk. Our results imply that the formation of the blastoderm disc involves the aggregation of cells at the posterior pole of the egg and not the migration of nuclei within a syncytial cytoplasm. These findings suggest that the primary cleavage syncytium does not play the same role in patterning the locust embryo as it does in Drosophila. However, we do identify a syncytial nuclear layer that underlies the forming blastoderm and remains in continuity with the yolk. PMID- 9226448 TI - Developmental potency of the murine allantois. AB - The murine allantois is the future umbilical component of the placenta. The base of the allantois is also thought to contain the future germ line. We have examined the fate and developmental potency of cells within the murine allantois during gastrulation. lacZ-expressing headfold-stage allantoises (approximately 8.0 days postcoitum; dpc) were subdivided into three proximodistal regions and transplanted into three sites in synchronous non-transgenic host embryos: the primitive streak at the level of prospective paraxial mesoderm, the primitive streak at the level of lateral plate mesoderm, and the base of the allantois. After 23 hours in culture, operated conceptuses were examined histologically for contribution of donor allantoic cells to the conceptus. None of the allantoic regions contributed to paraxial mesoderm when placed into the fetus, but all three colonized the endothelium and adjacent mesenchyme of the dorsal aorta. The mid-region was most efficient at colonizing endothelium, whereas the base was the only allantoic region to exhibit relative pluripotency, colonizing several derivatives of all three primary germ layers. Differences in the state of differentiation along the proximodistal axis of the allantois were further borne out when the three allantoic regions were placed into the base of the allantois of host conceptuses. Striking differences were observed in final position along the proximodistal axis of the host allantois. Most grafted cells translocated distally from the base; however, basal donor allantoic cells translocated typically only as far as the host's mid-region, whereas donor allantoic tip cells typically returned to the tip, often colonizing the chorioallantoic fusion junction. Together, our data reveal that the headfold-stage allantois may contain a proximodistal gradient of differentiation, and raise intriguing questions about how this gradient was established and the role it plays in umbilical vasculogenesis. PMID- 9226449 TI - A novel prespore-cell-inducing factor in Dictyostelium discoideum induces cell division of prespore cells. AB - In Dictyostelium discoideum strain V12M2, at a very low cell density (approximately 10(2) cells/cm2), most amoebae differentiate into prespore cells in a salt solution containing cAMP if an adequately diluted conditioned medium (CM) is provided (Oohata, A. A. (1995) Differentiation 59, 283-288). This finding suggests the presence of factor(s) released into the medium that are involved in inducing prespore cell differentiation. In the present study, we report the presence of two types of factors that function synergistically in prespore cell induction; one is a heat-stable and dialysable factor(s) and the other is a heat labile and non-dialysable factor termed psi (psi) factor (prespore-inducing factor). We purified and characterized the psi factor. Its relative molecular mass was determined to be 106x10(3) Mr by SDS-PAGE and 180x10(3) Mr by gel filtration HPLC, respectively. These results indicate that psi factor exists as a dimer under native conditions. In addition to inducing prespore cell differentiation, psi factor induced cell division of prespore cells in submerged culture. Our results suggest that psi factor plays important roles not only in prespore cell differentiation but also in the progress of the cell cycle in the prespore pathway in normal development. PMID- 9226450 TI - Infection of the germ line by retroviral particles produced in the follicle cells: a possible mechanism for the mobilization of the gypsy retroelement of Drosophila. AB - The gypsy retroelement of Drosophila moves at high frequency in the germ line of the progeny of females carrying a mutation in the flamenco (flam) gene. This high rate of de novo insertion correlates with elevated accumulation of full-length gypsy RNA in the ovaries of these females, as well as the presence of an env specific RNA. We have prepared monoclonal antibodies against the gypsy Pol and Env products and found that these proteins are expressed in the ovaries of flam females and processed in the manner characteristic of vertebrate retroviruses. The Pol proteins are expressed in both follicle and nurse cells, but they do not accumulate at detectable levels in the oocyte. The Env proteins are expressed exclusively in the follicle cells starting at stage 9 of oogenesis, where they accumulate in the secretory apparatus of the endoplasmic reticulum. They then migrate to the inner side of the cytoplasmic membrane where they assemble into viral particles. These particles can be observed in the perivitelline space starting at stage 10 by immunoelectron microscopy using anti-Env antibodies. We propose a model to explain flamenco-mediated induction of gypsy mobilization that involves the synthesis of gypsy viral particles in the follicle cells, from where they leave and infect the oocyte, thus explaining gypsy insertion into the germ line of the subsequent generation. PMID- 9226451 TI - Positional apoptosis during vertebrate CNS development in the absence of endogenous retinoids. AB - We have previously shown that quail embryos that develop in the absence of vitamin A have severe defects in their central nervous system. One defect is a completely missing posterior hindbrain. Here we have studied how this comes about by examining cell death using a wholemount technique. In these A- embryos we observe two narrow bands of ectopic apoptosis. One is in the mesenchyme in the region of the first somite and occurs at the 4-6 somite stage, before neural tube closure. The second band follows immediately afterwards and occurs in the neuroepithelium of the presumptive posterior hindbrain at the 6-8 somite stage. Electron microscopy shows that the dying neuroepithelial cells exhibit the characteristics of apoptosis. Rescuing the embryos by injecting retinol before gastrulation completely prevents these apoptotic events. In an effort to identify some of the genes that may be involved in the apoptotic pathway we show that Msx 2 is upregulated in the apoptotic neuroepithelium and thus may be involved, whereas Bmp-4 is not altered and thus presumably not involved. Since these apoptotic event take place at the time of specification of axial identity and segmentation in the mesenchyme and neuroepithelium we conclude that these cells die because they are wrongly specified in terms of their rostrocaudal position, a novel phenomenon which we refer to as positional apoptosis. PMID- 9226452 TI - The bicoid-related homeoprotein Ptx1 defines the most anterior domain of the embryo and differentiates posterior from anterior lateral mesoderm. AB - Ptx1 is a member of the small bicoid family of homeobox-containing genes; it was isolated as a tissue-restricted transcription factor of the pro-opiomelanocortin gene. Its expression during mouse and chick embryogenesis was determined by in situ hybridization in order to delineate its putative role in development. In the head, Ptx1 expression is first detected in the ectoderm-derived stomodeal epithelium at E8.0. Initially, expression is only present in the stomodeum and in a few cells of the rostroventral foregut endoderm. A day later, Ptx1 mRNA is detected in the epithelium and in a streak of mesenchyme of the first branchial arch, but not in other arches. Ptx1 expression is maintained in all derivatives of these structures, including the epithelia of the tongue, palate, teeth and olfactory system, and in Rathke's pouch. Expression of Ptx1 in craniofacial structures is strikingly complementary to the pattern of goosecoid expression. In addition, Ptx1 is expressed early (E6.8) in posterior and extraembryonic mesoderm, and in structures that derive from these. The restriction of expression to the posterior lateral plate is later evidenced by exclusive labelling of the hindlimb but not forelimb mesenchyme. In the anterior domain of expression, the stomodeum was shown by fate mapping to derive from the anterior neural ridge (ANR) which represents the most anterior domain of the embryo. The concordance between these fate maps and the stomodeal pattern of Ptx1 expression supports the hypothesis that Ptx1 defines a stomodeal ectomere, which lies anteriorly to the neuromeres that have been suggested to constitute units of a segmented plan directing head formation. PMID- 9226453 TI - Genetic basis of the formation and identity of type I and type II neurons in Drosophila embryos. AB - The embryonic peripheral nervous system of Drosophila contains two main types of sensory neurons: type I neurons, which innervate external sense organs and chordotonal organs, and type II multidendritic neurons. Here, we analyse the origin of the difference between type I and type II in the case of the neurons that depend on the proneural genes of the achaete-scute complex (ASC). We show that, in Notch- embryos, the type I neurons are missing while type II neurons are produced in excess, indicating that the type I/type II choice relies on Notch mediated cell communication. In contrast, both type I and type II neurons are absent in numb- embryos and after ubiquitous expression of tramtrack, indicating that the activity of numb and the absence of tramtrack are required to produce both external sense organ and multidendritic neural fates. The analysis of string embryos reveals that when the precursors are unable to divide they differentiate mostly into type II neurons, indicating that the type II is the default neuronal fate. We also report a new mutant phenotype where the ASC-dependent neurons are converted into type II neurons, providing evidence for the existence of one or more genes required for maintaining the alternative (type I) fate. Our results suggest that the same mechanism of type I/type II specification may operate at a late step of the ASC-dependent lineages, when multidendritic neurons arise as siblings of the external sense organ neurons and, at an early step, when other multidendritic neurons precursors arise as siblings of external sense organ precursors. PMID- 9226454 TI - Chimeric analysis of fibroblast growth factor receptor-1 (Fgfr1) function: a role for FGFR1 in morphogenetic movement through the primitive streak. AB - Fibroblast growth factor (FGF) signaling has been implicated in the patterning of mesoderm and neural lineages during early vertebrate development. In the mouse, FGF receptor-1 (FGFR1) is expressed in an appropriate spatial and temporal manner to be orchestrating these functions. Mouse embryos homozygous for a mutated Fgfr1 allele (fgfr1(delta tmk)) die early in development, show abnormal growth and aberrant mesodermal patterning. We have performed a chimeric analysis to further study FGFR1 function in the morphogenesis and patterning of the mesodermal germ layer at gastrulation. At E9.5, fgfr1(delta tmk)/fgfr1(delta tmk) cells showed a marked deficiency in their ability to contribute to the extra-embryonic, cephalic, heart, axial and paraxial mesoderm, and to the endoderm of chimeric embryos. Analysis at earlier stages of development revealed that fgfr1(delta tmk)/fgfr1(delta tmk) cells accumulated within the primitive streak of chimeric embryos, and consequently failed to populate the anterior mesoderm and endodermal lineages at their inception. We suggest that the primary defect associated with the fgfr1(delta tmk) mutation is a deficiency in the ability of epiblast cells to traverse the primitive streak. fgfr1(delta tmk)/fgfr1(delta tmk) cells that accumulated within the primitive streak of chimeric embryos tended to form secondary neural tubes. These secondary neural tubes were entirely fgfr1(delta tmk)/fgfr1(delta tmk) cell derived. The adoption of ectopic neural fate suggests that normal morphogenetic movement through the streak is essential not only for proper mesodermal patterning but also for correct determination of mesodermal/neurectodermal cell fates. PMID- 9226456 TI - Myocardial perfusion after cholesterol lowering. PMID- 9226455 TI - Goosecoid and HNF-3beta genetically interact to regulate neural tube patterning during mouse embryogenesis. AB - The homeobox gene goosecoid (gsc) and the winged-helix gene Hepatic Nuclear Factor-3beta (HNF-3beta) are co-expressed in all three germ layers in the anterior primitive streak and at the rostral end of mouse embryos during gastrulation. In this paper, we have tested the possibility of functional synergism or redundancy between these two genes during embryogenesis by generating double-mutant mice for gsc and HNF-3beta. Double-mutant embryos of genotype gsc(-/-);HNF-3beta(+/-) show a new phenotype as early as embryonic days 8.75. Loss of Sonic hedgehog (Shh) and HNF-3beta expression was observed in the notochord and ventral neural tube of these embryos. These results indicate that gsc and HNF-3beta interact to regulate Shh expression and consequently dorsal ventral patterning in the neural tube. In the forebrain of the mutant embryos, severe growth defects and absence of optic vesicles could involve loss of expression of fibroblast growth factor-8, in addition to Shh. Our results also suggest that interaction between gsc and HNF-3beta regulates other signalling molecules required for proper development of the foregut, branchial arches and heart. PMID- 9226457 TI - Using genetically modified mice to study apolipoprotein B. AB - The B apolipoproteins, apo-B48 and apo-B100, are key proteins in mammalian lipoprotein metabolism and are components of all classes of lipoproteins considered to be atherogenic. Our laboratory has generated an array of genetically modified mice for studying apo-B biology. Using gene targeting in mouse embryonic stem cells, we have generated apo-B-deficient mice. Heterozygotes had low plasma levels of apo-B and cholesterol; homozygotes died early in embryonic development, most likely because the absence of lipoprotein secretion by the yolk sac interfered with the delivery of lipid nutrients to the developing embryo. We have also generated human apo-B transgenic mice with an 80-kb genomic DNA fragment spanning the entire human apo-B gene; those mice had markedly increased plasma levels of low density lipoprotein cholesterol and exhibited increased susceptibility to atherosclerosis. The human apo-B transgenic mice have also yielded insights regarding the regulation of apo-B expression in different tissues. Although the 80-kb transgene contained nearly 20 kb of 5' and 3' flanking sequences and was expressed at high levels in the liver, no transgene expression was detectable in the intestine. Subsequent transgenic mouse studies have demonstrated that the expression of the apo-B gene in the intestine is controlled by DNA sequences that are very distant from the structural gene. Transgenic mice have also proved useful for studying apo-B structure/function relationships. By expressing mutant forms of human apo B in transgenic mice, we have examined the structural features of the apo-B molecule that are required for lipoprotein (a) formation. We have demonstrated that the carboxyl terminal cystine residue of apo-B100, cysteine-4326, is required for apo-B100's disulfide linkage with apo(a) to form lipoprotein (a). Finally, we have used gene targeting techniques to generate mice that synthesize exclusively apo-B48 (apo B48-only mice) and mice that synthesize exclusively apo-B100 (apo-B100 only mice): These mice have helped to clarify the unique metabolic roles of the two apo-B proteins. PMID- 9226458 TI - Modulation of endothelial cell apoptosis: mechanisms and pathophysiological roles. AB - Apoptosis is a mode of cell death in which intrinsic cellular mechanisms participate in the demise of the cell. The modulation of endothelial apoptosis may play a role in atherosclerosis, angiogenesis, vascular remodeling and other pathophysiological states. Control of cell death is mediated by the state of activation of a death pathway as well as by the levels of anti apoptotic proteins. The final common pathway of many, if not all, triggers of apoptosis involves activation of cysteine proteases. The Bcl 2 family, in contrast, appears to play a major role in protection against apoptosis. The role of these mechanisms in modulating endothelial cell apoptosis under various conditions is discussed. PMID- 9226459 TI - Regulation of triglyceride metabolism by PPARs: fibrates and thiazolidinediones have distinct effects. AB - The molecular mechanism by which hypolipidemic fibrates and antidiabetic thiazolidinediones exert their hypotriglyceridemic action are discussed. Increased activity of lipoprotein lipase (LPL), a key lipolytic enzyme, and decreased levels of apolipoprotein C-III (apo C-III) seem to explain the hypotriglyceridemic effects of compounds. Both fibrates and thiazolidinediones exert their action by activating transcription factors of the peroxisome proliferator activated receptor (PPAR) family, thereby modulating the expression of the LPL and apo C-II genes. First, treatment of rats with PPAR alpha activators, such as fibrates induced LPL mRNA and activity selectively in the liver. In contrast, the thiazolidinediones, which are high affinity ligands for PPAR gamma, have no effect on liver, but induce LPL mRNA and activity levels in adipose tissue. In hepatocytes, fibrates, unlike the thiazolidinediones, induce LPL mRNA levels, whereas in preadipocyte cell lines the PPAR gamma ligand induces LPL mRNA levels much quicker and to a higher extent than fibrates. Second, apo C III mRNA and protein production strongly decrease in livers of fibrate but not thiazolidinedione-treated animals. Fibrates also reduced apo C-III production in primary cultures of rat and human hepatocytes. The modulation of the expression of the LPL and apo C-III genes by either PPAR alpha or gamma activators, correlates with the tissue-specific distribution of the respective PPARs: PPAR gamma expression is restricted to adipose tissues, whereas PPAR alpha is expressed predominantly in liver. In both the LPL and apo C-III genes, sequence elements responsible for the modulation of their expression by activated PPARs have been identified which supports that the transcriptional regulation of these genes by fibrates and thiazolidinediones contributes significantly to their hypotriglyceridemic effects in vivo. Whereas thiazolidinediones predominantly affect adipocyte LPL production through activation of PPAR gamma, fibrates exert their effects mainly in the liver via a PPAR alpha-mediated reduction in apo C III production. This tissue specific transcriptional regulation of genes involved in lipid metabolism by PPAR activators and/or ligands might have important therapeutic implications. PMID- 9226460 TI - Role of molecular regulation in vascular calcification. AB - Calcium deposits account for most of the dry weight of atherosclerotic lesions. Previously considered uncommon, vascular calcification is now known to be present in 80% of significant lesions and in at least 90% of patients with coronary artery disease. Previously considered a passive process, it is increasingly recognized as an active, regulated process. Previously considered benign, it is now becoming recognized as a major risk factor for cardiovascular events, and a major contributor to systolic hypertension, heart failure, plaque rupture and stenosis. To confirm the similarity of vascular calcification with embryonic osteogenesis, we demonstrated the expression of bone morphogenetic protein in calcified human lesions, and we developed an in vitro model of vascular calcification that provides a useful experimental system for elucidating the molecular regulation of this process, which we have shown to include alkaline phosphatase induction and expression of bone matrix proteins and differentiation factors. Understanding the regulatory mechanisms of vascular calcification will allow future therapeutic approaches to prevent and possibly reverse this disease and its clinical consequences. PMID- 9226461 TI - A coronary primary intervention study of Japanese men: study design, implementation and baseline data. The Kyushu Lipid Intervention Study Group. AB - This report describes the design and baseline results of the Kyushu Lipid Intervention Study (KLIS). The study aims to test the hypothesis that the long term reduction of serum total cholesterol by pravastatin will lead to a decrease in coronary heart disease (CHD) events. The trial was designed to include a random 6,000 male patients aged 45-74 years with serum total cholesterol of 220 mg/dl (5.69 mmol/l) or greater and without a history of myocardial infarction, coronary surgery or angioplasty, to undertake either pravastatin or conventional treatment (including hypolipidemic drugs other than HMG CoA reductase inhibitors, probucol and bezafibrate), and to follow up each patient for 5 years. Primary endpoints are fatal and nonfatal myocardial infarction, coronary bypass surgery and angioplasty, cardiac death, and sudden and unexpected death. During the period from May 1990 to September 1993, a total of 5,640 male patients aged 45-74 were recruited by 902 participating physicians throughout Kyushu. Randomization was, however, neglected by study physicians; the numbers of patients enrolled were 3,061 in the pravastatin group and 2,579 in the conventional treatment group. Patients allocated to the pravastatin treatment were generally unfavorable regarding coronary risk factors. Baseline mean levels of serum total cholesterol were 259 mg/dl (6.70 mmol/l) in the pravastatin group and 246 mg/dl (6.36 mmol/l) in the conventional treatment group (p <0.001). Although the trial was regarded as a prospective observational study, the KLIS provides valuable quantitative data regarding cholesterol lowering and reduction in CHD events as well as safety data of the long-term use of a statin in Japanese men with hypercholesterolemia. PMID- 9226462 TI - Cholesterol levels among Japanese Americans and other populations: Seattle Nikkei Health Study. AB - The purpose of this study was to compare average cholesterol levels between Seattle based Japanese Americans and three other populations: U.S. population, native Japanese population and native Japanese urban workers. A total of 1,466 Japanese Americans (724 men and 742 women) participated in cardiovascular disease screening in the Seattle area during 1989 94. Data sources for comparisons are from the Third National Health and Nutrition Examination Survey for 1988-91, the results of the National Cardiovascular Disease Examination Survey in Japan for 1990, and cardiovascular disease screening conducted by the Epidemiological Arteriosclerosis Research Institute in Japan for 1989. Total cholesterol and triglyceride levels of Seattle Japanese American men and women were highest among the four populations. Among men, high density lipoprotein cholesterol (HDL-C) levels for Seattle Japanese Americans and native Japanese were similar and fell between those of urban Japanese workers and the U.S. population. In women, the average HDL C levels were highest in the Japanese urban workers, second highest in Seattle Japanese Americans, and lowest in both the U.S. population and native Japanese population. These differences in lipid levels may be caused by both genetic and environmental factors, which are now under investigation. PMID- 9226464 TI - Fanconi's anaemia: case history of six Spanish families. AB - We report on the results obtained in 6 Fanconi's anaemia families (FA) (parents, brothers and sisters) affected by at least one of the symptoms usually observed in FA. The 6 FA families were studied from 1974 to 1990, all having located in Madrid (Spain) but with different ethnic origin: 3 families are of Spanish descent and the other 3 are gipsy families. All showed characteristics of the disease, including malformations, stunted growth, microcephaly, skin hyperpigmentation, high incidence of chromosomal breaks in lymphocyte cultures, and hematological and biochemical abnormalities: pancytopeny, increased fetal hemoglobin levels and significantly decreased superoxide dismutase (SOD) activity. (Ref. 17.) PMID- 9226463 TI - Effects of estrogen on atherosclerosis formation and serum nitrite/nitrate concentrations in cholesterol-fed ovariectomized rabbits. AB - This study was to examine the effects of estrogen replacement on atherosclerosis formation in ovariectomized cholesterol-fed rabbits. We also examined serum levels of nitrite/nitrate, stable metabolites of nitric oxide, to investigate the involvement of nitric oxide. Female New Zealand White rabbits were ovariectomized and divided into 3 groups; 1) fed a normal diet (ND group, n=5), 2) fed a 1% cholesterol diet (CD group, n=6), or 3) fed a 1% cholesterol diet and received estrogen replacement (CD+E group, n=7). After 3 months, the rabbits were sacrificed to examine atherosclerosis formation. Atherosclerosis was not observed in ND. The oil red 0 positive area in the aorta was significantly greater in CD than in CD+E (CD, 17.3+/-2.2; CD+E, 9.3+/-0.8%, p<0.05). Stenosis of the coronary artery was also significantly greater in CD than in CD+E (CD, 30.6+/-9.7; CD+E, 6.7+/-2.9%, p <0.05). There was no significant difference in serum lipids between CD and CD+E. Serum nitrite/nitrate levels were significantly lower in CD than in ND (ND, 37.6+/-3.6; CD, 25.3+/-3.1 microM, p<0.05). There was a non-significant trend towards higher nitrite/nitrate levels after estrogen replacement (CD+E, 34.4+/-3.8 microM, p=0.08 vs. CD). These results suggest that direct actions on vascular wall including nitric oxide production contribute to the anti atherogenic effects of estrogen. PMID- 9226465 TI - [Development of high-throughput polymerase chain reaction system and its performance]. AB - We have developed a high-throughput thermalcycler that allows reduction in reagent volume and large number of PCR amplifications. One microliter of PCR samples can be amplified on the 384-well Teflon-coated aluminum plate. Four plates can be simultaneously subjected to PCR reaction in one instrument (1536 reactions). The heated lid is continually regulated at 2-5 degrees C higher temperature than samples during PCR cycle. This enable to prevent the evaporation in small reaction volume without use of mineral oil. Also, this PCR system employs the special carbongraphite sheet showing high thermal conductivity, resulting in rapid, uniform and reproducible amplification. Furthermore, all system including temperature regulation and open-close of the lids can be automatically controlled by personal computer. The number of the machines which can be synchronously controlled in one integrated PCR system is expanded to achieve the required throughput of PCR reactions. In this paper, we demonstrate that this instrument greatly facilitates to perform a large number of PCR reactions, to reduce the running cost and accelerates the computer-controlled PCR robotization. PMID- 9226466 TI - [Functional role of muscarinic M1 receptor for the ganglionic transmission in the rat superior cervical ganglion]. AB - To elucidate the sympathetic ganglia transmission via muscarinic M1 receptor subtype, we focused on the external carotid nerve (ECN), which branches from the superior cervical ganglion and innervates the thyroid gland. In addition, thyroid blood flow (TBF) was measured by a Laser Doppler blood flow meter as an indicator for the function of ECN. A relatively specific M1 agonist, AF102B, elicited a burst depolarization of ECN and an increase in TBF. Pretreatment with a selective M1 antagonist, pirenzepine, inhibited these responses. Superior cervical ganglionectomy also suppressed the AF102B-induced increase in TBF. In contrast, electric stimulation of the sympathetic trunk elicited a TBF decrease. Nicotinic receptor agonist, DMPP (dimethylphenylpiperazinium) evoked a short-term ECN depolarization, but decreased the TBF. These responses were blocked by nicotinic receptor antagonist, hexamethonium (C6), but not only by pirenzepine. Pretreatment with nitric oxide (NO) synthase inhibitor, L-NAME, suppressed the AF102B-induced increase in TBF. These findings suggest that the M1 receptor subtype may modulate the sympathetic ganglionic transmission which has a mechanism different from nicotinic transmission in terms of functional roles, i.e., blood flow changes. Furthermore, the NO system might be involved in sympathetic ganglia transmission via the M1 receptor subtype in the rat cervical ganglion. PMID- 9226467 TI - [Augmentation of vaccination effects of PGE2-producing tumor cells by transfection with cytokine genes]. AB - Potentially antigenic tumors often escape from the immune surveillance system by producing immunosuppressive factors. It has previously been reported that a progressor-type murine fibrosarcoma culture line, QRpP cells, produced high levels of PGE2 compared with a regressor-type tumor line, QR-32 cells, and that QRpP cells progressively grew in syngeneic C57BL/6 mice. In order to improve suppressed immunogenicity of QRpP cells with the high levels of PGE2, the author transfected QRpP cells with an effective expression vector containing cDNA for IL 2, IFN-gamma and TNF-alpha. These transfected clones expressed mRNA for IL-2, IFN gamma and TNF-alpha, respectively, and produced high levels of cytokines in their culture supernatants. Consequentry, QRpP cells transfected with IL-2 (QRpP-IL-2), IFN-gamma (QRpP-IFN-gamma), IL-2 and IFN-gamma (QRpP-IL-2/IFN-gamma), TNF-alpha (QRpP-TNF-alpha) lowered in tumorigenicity. In particular, the mice that had rejected the implantation of QRpP-IL-2 clone also rejected implantation of the parental QRpP cells. The activity of pre-cytotoxic T cells (pre-CTLs) obtained from the mice implanted with QRpP-IL-2 clone was higher than that of the mice implanted with other transfectants. These results suggested that transfection with the gene for IL-2 is an effective strategy against the producing immunosuppressive factors such as PGE2. PMID- 9226468 TI - [Defective generation of NK1.1+ T cells in aly/aly mice associated with thymic architecture]. AB - It has been reported that positive selection of natural killer antigen 1.1+ (NK1.1+) T cell antigen receptor (TCR) alpha beta+ thymocytes recently identified among CD4+8- and CD4-8- subpopulations is attributable to major histocompatibility complex (MHC) class Ib ligands expressed on bone marrow (BM) derived components in the thymus. This selection pattern is quite different from NK1.1-T cells of main stream. In the present study, we investigated generation of NK1.1+ TCR alpha beta+ cells in the thymus of aly/aly mouse which lacks lymph nodes and Peyer's patches and shows abnormalities of thymic and splenic structure. We found that the proportion of the NK1.1+ TCR alpha beta+ thymocytes was extremely low in these mice as compared with aly/aly+ and normal C57BL/6 mice. Thymic reconstitution by BM cells from aly/aly+ mice which possess a normal population of NK1.1+ TCR alpha beta+ thymocytes did not restore the NK1.1+ TCR alpha beta+ cell population in the thymus of lethally irradiated aly/aly mouse. When deoxy-guanosine (dGuo)-treated fetal thymi from (B6 x B10.G) F1 mice were transplanted to aly/aly mice which had been thymectomized and reconstituted with BM cells of aly/aly mice, normal proportions of the NK1.1+ TCR alpha beta+ thymocytes were observed in the thymus grafts. Furthermore it was demonstrated that NK1.1+ T cells in aly/aly mice were unable to produce efficiently IL-4 upon in vivo stimulation with anti-CD3. These findings demonstrate that the development of NK1.1+ TCR alpha beta+ cells is accomplished under the influence of not only BM derived components but also in intact microenvironment of lymphoid tissues. PMID- 9226469 TI - [Evaluation on blood clearance and hepatic uptake of 99mTc-GSA in rats with blood flow conversion]. AB - This study was aimed to clarify a contribution of the hepatic blood flow in hepatic accumulation of 99mTc-DTPA-galactosyl serum albumin (99mTc-GSA). The experiment was performed in rats by the blood flow conversion with an external scintillation gamma camera and laser doppler flowmeter. Rats were divided into 4 groups: hepatic artery ligation (HAL, n = 10), portal vein ligation (PVL, n = 8), both hepatic artery and portal vein ligation (HAL+PVL, n = 9), and control (CONT, n = 10) groups. The scintigraphic data were obtained in each group for 10 minutes after intravenous injection of 99mTc-GSA (50 micrograms/100 g B.W). The regions of interest were assigned over the heart and whole liver and the time activity curves (TAC) were generated. Five parameters of HH 4, LHL 4, KH1, KH2, KL, were calculated as blood clearance and hepatic accumulation from TAC in each rat. HH4 as blood clearance index in CONT, HAL, PVL and HAL+PVL was 0.58 +/- 0.04 (mean +/ SE), 0.63 +/- 0.04, 0.85 +/- 0.04, 0.97 +/- 0.001, respectively. HH 4 between CONT vs PVL, HAL+PVL was statistically significant (p < 0.05). LHL 4 as hepatic uptake index in CONT, HAL, PVL and HAL+PVL was 0.96 +/- 0.001, 0.93 +/- 0.01, 0.71 +/- 0.07, 0.41 +/- 0.04, respectively. This parameter was also statistically significant between CONT vs PVL and HAL+PVL groups. Another parameter of KH1 for blood clearance and KL for hepatic uptake were also significant between CONT vs PVL and HAL+PVL groups. All parameters obtained in 99mTc-GSA study correlated well with the hepatic flow rate which was measured with a laser doppler flowmeter and reflected the reduction rate of the hepatic tissue blood flow 4 minutes after the ligation of target vessels. These result suggest that blood clearance and hepatic uptake of 99mTc-GSA are significantly affected by hepatic blood inflow. 99mTc-GSA scintigraphy may be useful in evaluating hepatic tissue blood flow. PMID- 9226470 TI - [The essential region of CD40 cytoplasmic domain for signal transduction in WEHI231 cells]. AB - Ligation of surface immunoglobulin M (IgM) induced cell death and this was blocked by CD40-mediated signal in a mouse B cell line, WEHI231. In order to define which region of CD40 and how CD40-mediated signal are involved in the blocking of IgM-induced cell death, we generated several human CD40 transfectants. We demonstrated that a 10-amino acid segment in the conserved cytoplasmic region of CD40-mediated rescue from IgM-induced cell death and was involved in the activation of c-Jun aminoterminal kinase (JNK). In addition, the same segment was also important for CD40-mediated activation of extracellular signal-regulated protein kinase 2 (ERK2). Moreover, tumor necrosis factor receptor-associated factor (TRAF) 2 and 3 could bind to the same segment of CD40. PMID- 9226471 TI - [Frequency and effect on plasma lipoprotein metabolism of a mutation in the cholesteryl ester transfer protein gene in the Chinese]. AB - The aspartate to glycine substitution at codon 442 (D442G) in the cholesteryl ester transfer protein (CETP) gene is a common mutation in Japan and is thought to be a factor that elevates the serum high-density lipoprotein (HDL)-cholesterol level in the general Japanese population. In the present study, we investigated the frequency of the D442G mutation in the Chinese subjects living in Beijing. We also studied the effects of the mutation on plasma lipoprotein metabolism in the same population and compared it with that in the Japanese Diagnosis of D442G mutation was established by polymerase chain reaction followed by restriction digestion. Among 379 subjects studied, 16 were found heterozygous thereto (allelic frequency = 2.1%). Unexpectedly, the D442G mutation was not associated with elevated HDL-cholesterol levels in the Chinese. Instead, in comparison to the unaffected, significantly decreased low-density lipoprotein (LDL)-cholesterol and total cholesterol levels were observed in the heterozygotes. These results were discrepant from those obtained in Japan: Japanese D442G heterozygotes had elevated HDL-cholesterol levels with unchanged LDL-cholesterol and total cholesterol levels. The divergence in the effect of D442G mutation on plasma lipoprotein metabolism appears to be related to the significantly low CETP activities in the Chinese affected and unaffected. Although the difference between Japan and China in nutritional conditions is supposed to contribute to the divergence, the precise mechanism remains to be determined. PMID- 9226472 TI - [In vitro evaluation of metabolic change in forebrain ischemia model of rat using proton magnetic resonance spectroscopy]. AB - Metabolic disruption resulted from cerebral ischemia and post-ischemia reperfusion injury was studied using proton magnetic resonance spectroscopy (1H MRS). We also analyzed the effect of 3-methyl-1-phenyl-2-pyrazolin-5-one (MCI 186) which can scavenge free radicals induced in the brain tissue due to ischemic reperfusion in this experiment. The ischemic model was produced using rat forebrain ischemic model (Pulsinelli's 4 vessels occlusion model). Post-ischemic reperfusion was also induced by the re-opening of the occluded common carotid arteries. The occluded time was 30 min and reperfusion time 0, 10, 30, 60 min. We obtained the specimens in the cortex under microwave fixation. Choline and acetate increased during ischemia and gradually decreased during reperfusion period. These two signals seen in 1H MRS are supposed to represent cell membrane components (products) and the increase of these signals after reperfusion seems to be related to the post ischemic reperfusion injury due to the explosive increase of free radicals. Lactate, which is induced by anaerobic glycolysis, increased during ischemia and promptly disappeared after reperfusion. The treatment of pre-ischemic administration of MCI-186 significantly suppressed increases of choline and acetate. As far as lactate is concerned, post-ischemic administration of this drug significantly reduced its increase at the point of reperfusion. Our results suggest that MCI-186 alternates changes induced by ischemic-reperfusion injury in membranous metabolism, probably due to its free radical scavenging action. PMID- 9226473 TI - Biodistribution of synthetic thymosin beta 4 in the serum, urine, and major organs of mice. AB - Thymosin beta 4 (T beta 4) is a peptide of 43 amino acids that was first isolated from the thymus gland and subsequently found to be ubiquitous in nature. T beta 4 functions mainly as an actin-sequestering molecule in nonmuscle cells, where its primary role is to maintain the large pool of unpolymerized G-actin in the cell. Studies on the pharmacokinetics of T beta 4 in human and other mammals have not been reported so far. In the present study, we have measured T beta 4 concentrations in serum, urine, and 10 major organs of female Swiss-Webster mice following intraperitoneal administration of 400 micrograms synthetic T beta 4. Using a modified enzymatic immunoassay, our data show a significant increase of T beta 4 in serum starting 2 min after injection and lasting for 40 min (average: 2.34 +/- 0.54 micrograms/ml). High concentrations were found in urine (59.3 +/- 7.54 micrograms/ml) at three different points after injection (20 min, 40 min, and 2 h). Of the 400 micrograms T beta 4 administered to mice 83% was recovered at the end of the study, 44.6% of which corresponded to urine, 1.4% to serum, and 37.5% to the organs. In 50% of the tested organs, the wet weight concentrations of T beta 4 increased significantly from the first 40 min to 2 h after injection in comparison to their baseline wet weight concentrations. These organs were: the brain (72 micrograms/g), heart (80 micrograms/g), liver (15 micrograms/g vs 9 micrograms/g), kidneys (65 micrograms/g vs 28 micrograms/g), and peritoneal fat (47 micrograms/g vs 13 micrograms/g). Wet weight concentrations increased in the thymus (196 micrograms/g vs 147 micrograms/g) and muscle (45 micrograms/g vs 0 micrograms/g) after 6 h of injection. The spleen showed an increase in wet weight concentrations at the 2 min timepoint (267 micrograms/g vs 161 micrograms/g). Ovaries had a biphasic increase at 40 min (72 micrograms/g vs 62 micrograms/g) and 24 h (92 micrograms/g vs 62 micrograms/g) after T beta 4 administration. In lungs, the highest wet weight increase after injection (149 micrograms/g at timepoint 6 h) was not higher than its basal wet weight concentration (153 micrograms/g). These pharmacokinetic studies of T beta 4 in mice have established that high levels of T beta 4 are found in blood following I.P. administration and the kidney rapidly removes the peptide from the circulation. The kinetics of this response should help define the proper scheduling of administration of T beta 4 during clinical trials in disorders, such as the acute respiratory distress syndrome (ARDS), associated with actin toxicity. PMID- 9226474 TI - An extract of the fern Polypodium leucotomos inhibits human peripheral blood mononuclear cells proliferation in vitro. AB - An alcoholic extract of the fern polypodium leucotomos (PLE) has been empirically used as an immunosuppressor for the treatment of several autoimmune diseases. In this paper, we investigated the effects of PLE on activation and proliferative responses of peripheral blood mononuclear cells (PBMNC) from healthy donors to T lymphocyte polyclonal mitogens. PLE shows a significant inhibitory effect on the proliferative response of PBMNC to stimulation with phytohaemagglutinin (PHA) or anti CD3 monoclonal antibodies (p < 0.05). In contrast, PLE did not modify the proliferative response of PBMNC to phorbol esters (p > 0.05). The inhibitory effect of PLE upon mitogen induced PBMNC proliferation is time dependent and can be overcome by the exogenous addition of interleukin-2 to the culture medium (p < 0.05). The decreased proliferative response of PBMNC to PHA stimulation in the presence of PLE is not associated with a significant modification of expression of the alpha chain (CD25) of the IL-2 receptor (p > 0.05). In conclusion, PLE shows an inhibitory effect on the polyclonal proliferative response of PBMNC to T lymphocyte mitogens that interact with cytoplasmic membrane molecules. PMID- 9226475 TI - Suppression of arthritis by the inhibitors of dipeptidyl peptidase IV. AB - Dipeptidyl peptidase IV (DP IV, CD26) is a serine exoprotease which selectively cleaves the penultimate proline residue of polypeptides. This enzyme is also expressed as a surface marker on activated T cells. In order to assess the relevance of DP IV in immunological disorders, we evaluated the in vivo effects of specific DP IV inhibitors using two arthritis models, one which was induced by collagen one by alkyldiamine. These animal models share several pathological features associated with rheumatoid arthritis. The transition state substrate analog of DP IV, (S)-Alanylpyrrolidine-boronic Acid (Ala-boroPro), suppressed hind paw swelling, which was associated with collagen-induced and alkyldiamine induced arthritis. A competitive inhibitor of DP IV, Lys(Z(NO2))-thiazolidide and an irreversible inhibitor, Ala-Pro-nitrobenzoylhydroxylamine also suppressed alkyldiamine-induced arthritis dose-dependently. We also analyzed the pharmacological effects of Lys(Z(NO2))-thiazolidide on several immune responses in vitro, in order to determine its mode of action. This inhibitor suppressed mitogen-induced and antigen-induced proliferation of T cells. However, studies using splenic cells from DP IV deficient rats showed that the inhibition of lymphocyte proliferation was not exerted through the inhibition of DP IV. PMID- 9226476 TI - Effect of sciatic denervation on cell-mediated immunity. AB - Local semiallogenic graft-versus-host (GvH) and host-versus-graft (HvG) reactions were induced in mice whose right hind limbs had been partially denervated surgically. The denervation was performed by removing a 2 mm segment of the sciatic nerve, while sham-operated and non-operated mice served as controls. Eight days after denervation the mice were injected with spleen cells from a 3 month-old C57BL female, or from (C57xCBA)F1 male mice, and the GvH and HvG reactions were evaluated, 7 or 10 days later, by quantitatively estimating the weight (enlargement) of the popliteal lymph-nodes. Significant weight increases of 51% (for the GvH reaction) and 63% (for the HvG reaction) were observed in the denervated mice, when compared to the sham-operated ones. This increased cell type reactivity in a denervated limbs seems to be related to the immunostimulatory activity of cytokines which are released by cells engaged in repair processes of damaged nerves, as well as related to the reduction of the immunosuppressive effect of noradrenergic innervation in the denervated animals. PMID- 9226477 TI - Effects of interleukin-10 on water intake, locomotory activity, and rectal temperature in rat treated with endotoxin. AB - Intracerebroventricular (i.c.v.) interleukin-10 (25, 50, and 100 ng/rat) effects on water intake, exploratory behaviour, and rectal temperature were evaluated in rats treated intraperitoneally (i.p.) with lipopolysaccharide (0.32, 0.64, and 0.96 mg/kg). Endotoxin administration induced fever and inhibition of thirst in water-deprived rats, and a decrease in lococomotory activity in normohydrated and water-deprived animals. Our data show that interleukin-10 during lipopolysaccharide administration controlled fever, increases exploratory behaviour, but did not reverse lipopolysaccharide inhibition of thirst. These effects suggest that fever, depression in locomotory activity but not inhibition of thirst, induced by endotoxin are influenced by interleukin-10 levels. PMID- 9226478 TI - Analysis of thymic stromal cell subpopulations grown in vitro on extracellular matrix in defined medium--V. Proliferation regulating activities in supernatants of human thymic epithelial cell cultures. AB - In previous reports in this series, we described the growth conditions, morphology and supernatant activities of human thymic epithelial cell cultures. The human thymic epithelial cell supernatant (HTES) contained IL-6, G-CSF activities and exhibited a strong enhancing effect on thymocyte proliferative response to mitogens, which was identified as IL-6 related. The responding thymocyte population was apparently identified as PNA, mature T cells. In order to simplify further analysis of HTES activities, we selected to use a well defined mature murine T cell clone which has a Th2 phenotype (8-5 clone). HTES induced 8-5 cell proliferation without the presence of antigen, antigen presenting cells (APC) or mitogens. This enhancing effect of HTES was completely blocked with anti hIL-6 antibody but could not be reproduced by rhIL-6 alone. Hence, IL-6 is a necessary but insufficient factor in mediating this effect. HTES induced proliferation was accompanied by endogenous IL-4 secretion from 8-5 cells. Furthermore, the proliferation was blocked by anti mIL-4 antibody, implicating IL-4 as an autocrine growth factor in this system. HTES increased also the expression of IL-2 receptor. In addition, rhIL-2 and rmIL-4 each has a synergistic effect on the proliferative response of 8-5 cells to HTES. A similar synergistic activity was demonstrated when rhIL-6 was used instead of HTES, suggesting that IL-6 regulates some HTES activities. Our findings indicate that HTES activities, of which IL-6 is only part, are mediated via the induction of autocrine growth factors and by the regulation of T cell growth factor receptor expression. PMID- 9226479 TI - Vesnarinone inhibits immune-mediated but not Fas (CD95) agonist-mediated hepatic injury. AB - Previous studies have shown that the administration of concanavalin A (ConA) into mice induces immune-mediated liver injury, which can be largely abrogated by neutralizing tumor necrosis factor(TNF)alpha. Vesnarinone is an experimental drug which is known to inhibit TNF alpha release. Here we demonstrate that vesnarinone inhibits ConA-induced hepatic injury. In a dose-dependent manner, vesnarinone inhibits in several mouse strains the increase of serum aminotransferase concentrations. additional experiments show that vesnarinone inhibits ConA mediated accumulation of DNA fragmentation in the liver. Furthermore, the drug significantly reduces the levels of circulating TNF alpha and interleukin-6 (IL 6). Vesnarinone does not modulate TNF alpha and IL-6 action on hepatic cells, as shown by its failure to reduce the cytokine specific-stimulation of acute phase plasma proteins in the rat hepatoma H-35 cell line. Neither vesnarinone nor anti TNF alpha protect against direct liver injury induced by a sublethal dose of agonist anti-Fas (CD95) antibody. Taken together, these results suggest that vesnarinone blocks hepatic injury, in part by inhibiting the release of TNF alpha in vivo. PMID- 9226480 TI - Measurement and long-term health risks of child and adolescent fatness. AB - This paper reviews child and adolescent adiposity measures and associated long term health risks. The first section argues that anthropometric measures are practical for large scale epidemiological studies, particularly the body mass index. Limitations of this and other measures are presented. The second section summarises the evidence on the relationship between child and adolescent and adult adiposity. This is based on a search for relevant literature in the following computerised databases: Medline (1985-96), BIDS (EMBASE and Science Citation Index 1985-96). The literature search revealed that the child to adult adiposity relationship is now well-documented, although methodological differences hinder comparisons. Nonetheless, consistently elevated risks of adult obesity are evident for fatter children, although the prediction of adult obesity from child and adolescent adiposity measures is only moderate. Fewer studies could be identified in relation to long-term health risks of child and adolescent adiposity. It is therefore difficult to specify categories of risk associated with childhood adiposity without more information from long-term studies. Further evidence is also required to confirm the suggestion from some studies that adult disease risks are associated with a change in adiposity from normal weight in childhood to obesity in adulthood. However, on the basis of the evidence available, it is argued that population-based approaches to the prevention of obesity are likely to be more effective than approaches targeted as fat children. Population-based approaches are desirable, first because of the poor prediction of adult obesity from child and adolescent measures, and second, because of risks of adult mortality and morbidity may be elevated for individuals who become overweight after adolescence. PMID- 9226481 TI - Increase in both pro-thrombotic and anti-thrombotic factors in obese premenopausal women: relationship with body fat distribution. AB - OBJECTIVES: To examine the relationship of obesity, body fat distribution, and fasting plasma insulin concentrations with the plasma levels of both pro thrombotic and anti-thrombotic factors in premenopausal women. SUBJECTS: 32 obese women with BMI > 28 and 33 age-matched non-obese = women with BMI < 25. MEASUREMENTS: (i) plasma concentrations of plasminogen activator inhibitor-1 antigen (PAI-1 Ag), plasminogen activator inhibitor-1 activity (PAI-1 activity), fibrinogen, von Willebrand factor antigen (vWF Ag), von Willebrand factor activity (vWF activity), and factor VII activity as pro-thrombotic factors; (ii) plasma concentrations of tissue plasminogen activator antigen (t-PA Ag), protein C, and antithrombin III as anti-thrombotic factors; (iii) fasting plasma insulin and glucose concentrations, and the lipid pattern (triglycerides, total and HDL cholesterol) as metabolic parameters. The body fat distribution was evaluated by measuring the waist circumference and the waist-to-hip ratio (WHR). RESULTS: Obese subjects had higher plasma concentrations of all pro-thrombotic factors as compared to non-obese controls (PAI-1 Ag, P < 0.001; PAI-1 activity, P < 0.05; fibrinogen, P < 0.001; vWF Ag, P < 0.001; vWF activity, P < 0.05; factor VII, P < 0.05). The plasma concentrations of PAI-1 Ag and vWF Ag were directly correlated with the waist circumference independently of other metabolic and non-metabolic variables (P < 0.05). Obese women were also characterized by higher plasma concentrations of anti-thrombotic factors such as t-PA Ag and protein C as compared to non-obese controls (P < 0.001 and P < 0.001, respectively), although these factors were not independently correlated with the waist circumference or the WHR. CONCLUSION: Plasma concentrations of the pro-thrombotic factors are increased in obese women as compared to non-obese controls, and plasma levels of PAI-1 Ag and vWF Ag correlate with central fat accumulation specifically. Plasma concentrations of anti-thrombotic factors (namely protein C and t-PA Ag) are also raised in obese women, but they are not correlated with parameters of body fat distribution. The increase in protein C levels may represent a protective response partly counteracting the increase in pro-thrombotic factors in these individuals. PMID- 9226482 TI - Serum leptin concentration is associated with total body fat mass, but not abdominal fat distribution. AB - OBJECTIVE: The obese (ob) gene encodes leptin which inhibits appetite and stimulates thermogenesis. Serum leptin concentrations are determined by total body fat mass, but the influence of visceral fat accumulation and other metabolic factors have been clinically determined. METHODS: We determined the correlations between serum leptin concentrations and the total body fat mass, abdominal fat mass, abdominal fat distribution (estimated by ultrasound), and circulating metabolic factors in 104 Japanese healthy subjects (11 men and 93 women). In addition, the effect of food restriction (30 kcal/kg desired body weight/day) for four weeks on serum leptin concentrations were also examined in 30 women. RESULTS: There was a significant correlation between serum concentrations and total body fat mass (r = 0.708, P < 0.0001), the percentage of body fat (r = 0.561, P < 0.001), and the body mass index (BMI, r = 0.630, P < 0.001). Serum leptin concentrations were correlated with abdominal wall preperitoneal and subcutaneous fat pad thickness, but not the abdominal wall fat index (AFI). Serum leptin concentrations were also correlated with serum immunoreactive insulin (IRI), but not glucose, or free fatty acid (FFA) concentrations. The weight loss after food restriction for four weeks significantly (P = 0.016) reduced the serum leptin concentrations with a significant reduction of body fat mass, serum glucose, IRI and FFA concentrations. However, there was no significant correlation of the percentage change in serum leptin concentrations to that in body fat mass after food restriction. CONCLUSION: Serum leptin concentrations are well correlated with total body fat mass in healthy subjects. Differences in abdominal fat distribution do not appear to be related to a difference in the in vivo leptin production from adipose tissue. PMID- 9226483 TI - The reliability of upper limb anthropometry in older Chinese people. AB - OBJECTIVE: To evaluate the validity of the Durnin-Womersley equations and to derive our local predictive equations for body fat from upper limb skinfold thicknesses in older Chinese people in Hong Kong. To evaluate the validity of mid arm circumference and corrected arm muscle area in predicting lean tissue mass in the same population. DESIGN: Comparison of fat percentages predicted by Durnin Womersley (D-W) equations with those estimated by Dual energy X ray absorptiometry (DXA). Predictive equations derived from regression between upper limb skinfold thicknesses and fat percentages estimated by DXA were similarly evaluated in internal and external validation groups. Mid-arm circumference (MAC) and corrected arm muscle area (CAMA) were correlated with the limb lean tissue mass, body lean tissue mass and fat percentage. SUBJECTS: 354 female and 263 male, apparently well, community dwelling subjects, aged 69-82 y; of which 40 subjects of each sex were randomly selected from the study population for internal validation of the local predictive equations; 60 female and 33 male hospital medical outpatients, aged 61-87 y, were recruited for external validation. MEASUREMENTS: Triceps and biceps skinfold thicknesses, mid-arm circumference, body mass index, fat percentages, limb and whole body lean tissue masses estimated by Hologic QDR-2000 bone densitometer. RESULTS: Fat percentages calculated by D-W equations were significantly different from those estimated by DXA (average difference -2.4 (s.d. 4.8)% and +2.1 (5.2)% in females and males respectively). The corresponding differences for our local predictive equations were not significant (-0.9 (4.7)% and -0.5 (5.0)% in females and males respectively). There was a trend of under-estimation of body fat with increasing fatness. In the hospital medical outpatients, there was a significant difference between fat percentages predicted by our equation and those by DXA in female ( 2.9(5.3)%), but not in male (+0.3(4.3)%) subjects. In males, MAC correlated with limb and body lean tissue masses as well as with fat percentage (r = 0.60, 0.68, 0.65 respectively). CAMA correlated similarly well with lean tissue masses but was more independent of fat percentage (r = 0.61, 0.65, 0.44 respectively). In females, both MAC and CAMA correlated poorly with limb and body lean tissue masses. Moreover, MAC correlated well with fat percentage (r = 0.80). CONCLUSION: Upper limb skinfold thicknesses measurement is a valid means of predicting body fat in older Chinese people. Local predictive equations were more reliable that D W equations. They were, however, subject to errors at the extreme ends of body fatness and in the presence of disease. In older females, MAC and CAMA were not reliable in predicting lean tissue mass, but MAC could be used to predict fat percentages. In older males, CAMA was more reliable than MAC in predicting lean tissue mass. PMID- 9226484 TI - The effectiveness of long-term fibre supplementation on weight maintenance in weight-reduced women. AB - OBJECTIVE: To investigate whether fibre supplementation is effective in weight reduced subjects for maintenance of weight-loss in the long-term. DESIGN: Longitudinal, randomly assigned intervention study with supplementation of 20 g of water soluble fibre (guar gum) daily for 14 months after an energy-restricted period of two months (VLCD). SUBJECTS: Thirty-one female, obese subjects (age: 41.4 +/- 7.4 y: BMI 33.2 +/- 3.7 kg/m-2); 20 subjects were supplemented with fibre and 11 subjects served as the control group. MEASUREMENTS: Body weight (BW), blood lipids and blood pressure, anthropometry, and eating behaviour were measured before the VLCD (0), after VLCD (2), and at 4, 10, and 16 months. RESULTS: The fibre group with at least 80% compliance (group A) and the control group showed the same weight regain response after VLCD. The fibre consuming group with 50-80% compliance (group B) differed with respect to relapse. The rate and amount of BW regain was significantly higher for group B. After 14 months group B had returned to baseline levels, whereas group A and the control group showed a tendency to a lower BW than at baseline (P = 0.09). No effect of fibre supplementation was found on blood lipids, blood pressure and energy intake. Eating behaviour characteristics changed during the intervention and might explain differences in weight maintenance. CONCLUSIONS: No effect of 14 months fibre supplementation was found on weight maintenance in weight-reduced subjects. Guar gum intake did not result in reduction of blood pressure or cholesterol, or in suppression of energy intake. PMID- 9226485 TI - Association of poorly controlled diabetes with low serum leptin in morbid obesity. AB - OBJECTIVES: Leptin may be involved in the regulation of body weight, food intake, and energy expenditure. In view of a possible link between leptin concentrations and diabetes that has been suggested in obese rodents, we investigated the potential relationship between serum leptin concentrations and hyperglycaemia in French patients with morbid obesity. SUBJECTS: Fasting leptin concentrations were measured in 241 morbidly obese patients with various degrees of glucose tolerance in a cross-sectional study. RESULTS: Fasting serum leptin concentrations did not differ between normoglycaemia (NG, 61.5 +/- 24.0 ng/ml) and glucose intolerant morbidly obese subjects (IGT, 56.5 +/- 18.5 ng/ml) and were slightly lower in those with controlled diabetes (55.1 +/- 30.3 ng/ml, P = 0.06 when compared to NG subjects). In contrast, leptin concentrations were 30% lower in patients with poorly controlled diabetes (43.0 +/- 22.2 ng/ml, P = 0.001 vs NG subjects). Leptin concentrations were negatively correlated with fasting glucose in all groups combined (p = -0.24, P = 0.0001) and particularly in NIDDM subjects (p = 0.31, P = 0.0054). Although leptin concentrations were higher in women than in men, similar significant correlation with fasting glucose was found when females were analyzed separately. A positive correlation was found with BMI (p = 0.25, P = 0.0001) in all groups. Multivariate analysis revealed that fasting glucose was independently associated with serum leptin concentrations (F = 12.5, P = 0.0005). Sex, age, BMI, waist/hip ratio, fasting glucose and insulin, total cholesterol and triglycerides, tested in the model, explained 42% of the leptin variability in this population. CONCLUSIONS: Poorly controlled diabetes was accompanied by a significant reduction of serum leptin concentrations in morbidly obese subjects. We suggest that a relative leptin deficiency (lower than expected for the BMI) associated with insulin deficiency in this population might contribute to a vicious cycle maintaining (or even worsening) obesity itself and/or its metabolic complications. PMID- 9226487 TI - Determinants and nature of dietary underreporting in a free-living population: the Fleurbaix Laventie Ville Sante (FLVS) Study. AB - OBJECTIVE: To study the determinants and nature of dietary underreporting in a free-living population. DESIGN: Cross-sectional study of nutritional and behavioural characteristics. SUBJECTS: 1030 weight-stable subjects, 501 women and 529 men older than 15 y, included in the Fleurbaix Laventie Ville Sante study. MEASUREMENTS: Dietary intake was assessed using a 3 dy dietary record. Self assessed body weight and height were also recorded. Behavioural and socio economic data were obtained from a questionnaire. Underreporters were defined as people with a reported ratio of energy intake to estimated basal metabolic rate lower than 1.05. RESULTS: Underreporting concerned 16% of the population and was significantly more frequent in obese than in non obese subjects (P < 0.001). Underreporting was significantly associated with a high socio-professional class (P < 0.05), having dieted at least once (P < 0.01) and to be in dietary restraint (P < 0.05). Furthermore, the contribution of protein to energy intake was significantly higher in underreporters than in non underreporters, independently of weight status. CONCLUSIONS: These data underline that underreporting may bias the assessment of energy and macronutrient intake, particularly in studies on obesity and dietary restraint. Questions about weight concern, dieting and dietary restraint may be useful to identify subjects who underestimate their food intake. PMID- 9226486 TI - Effect of chronic intracerebroventricular infusion of insulin on brown adipose tissue activity in fed and fasted rats. AB - OBJECTIVES: Carbohydrate feeding stimulates, and fasting decreases the sympathetic nervous system activity and brown adipose tissue (BAT) thermogenesis. This study was performed to assess the hypothesis that these effects were secondary to changes in insulin concentrations in the central nervous system. METHODS: BAT sympathetic activity was assessed by comparing 3H-GDP binding to isolated mitochondria of innervated and denervated interscapular BAT of three groups of 10 week old male Wistar rats: food-restricted, 48 h fasted or ad libitum fed. During the three days preceding this measurement, animals received a continuous intracerebroventricular (ivc) infusion of insulin (0.48 U/d) or vehicle. RESULTS: In food-restricted rats, 3H-GDP binding to mitochondria of innervated BAT was 41% higher than that to denervated BAT. Icv insulin did not stimulate 3H-GDP binding in innervated BAT. In 48 h fasted rats, 3H-GDP binding to mitochondria of innervated BAT was reduced by 30-50%, while the activity of denervated BAT was minimally affected. Icv insulin did not prevent this fasting induced drop in BAT. In rats fed ad libitum, icv insulin decreased food intake by 17% (P < 0.05) and increased 3H-GDP binding to innervated BAT by 27% (P < 0.05). CONCLUSION: Intracerebroventricular insulin stimulates BAT activity in rats fed ad libitum but not in food-restricted or fasted rats. This demonstrates that the decrease in BAT activity observed during fasting is unlikely to be due to a decrease in insulin concentration in the nervous system. PMID- 9226488 TI - Long-term effects of a very low calorie diet (Nutrilett) in obesity treatment. A prospective, randomized, comparison between VLCD and a hypocaloric diet+behavior modification and their combination. AB - OBJECTIVES: To compare weight loss on a balanced hypocaloric diet to that of a Very Low Calorie Diet (VLCD) after two months of treatment and to further compare 26 months of weight maintenance and safety with or without VLCD assistance in obese patients. DESIGN: Prospective, randomized, controlled intervention trial, initially with two and later with three parallel groups. SETTING: Swedish University out-patient obesity clinic. SUBJECTS: Eighty-one obese patients of both gender with a BMI > or = 30 kg/m2 from the waiting list participated in a structured weight reduction + weight maintenance programme. INTERVENTION: Twenty seven patients (group A) were randomized to a balanced diet of 6720 kJ/d (1600 kcal/d) during the whole treatment period. The other patients (n = 54) were randomized to VLCD (Nutrilett) 1764 kJ/d (420 kcal/d) diet during the first two months. The VLCD treated patients were rerandomized after the initial treatment to the well balanced hypocaloric diet (6720 kJ/d) with (group C) or without (group B) 1 MJ of VLCD to be taken during the evening. MAIN OUTCOME MEASURES: During the first two-month period, the mean body weight loss in the VLCD group was 18.9 +/- 7.1 kg compared to 7.2 +/- 4.8 kg in the diet treated group, with a similar relative fat loss assessed by bioimpedance of 68% and 76% respectively. The maintained weight loss in all groups after 28 months of treatment was 10.9 +/ 10.2 kg in the 52% who completed the programme. Weight losses and drop-out rates were similar in all three groups. CONCLUSIONS: Twenty-four months weight maintenance and drop out rates are independent of whether the initial treatment commences with VLCD or a hypocaloric diet. One MJ nutrition powder taken freely does not affect 24 months weight maintenance on a hypocaloric (6.7 MJ/d) diet. PMID- 9226489 TI - Importance of intra-abdominal visceral fat accumulation to coronary atherosclerosis in heterozygous familial hypercholesterolaemia. AB - OBJECTIVES: Hyper-low-density lipoprotein (LDL)-cholesterolaemia is a potent risk factor for coronary atherosclerosis. We have recently demonstrated that a cluster of risk factors including insulin resistance, glucose intolerance, hypertriglyceridaemia, and hypertension based on intra-abdominal visceral fat accumulation are closely related to coronary artery disease. In the current study, we evaluated the relationship between visceral fat accumulation and the severity and distribution of coronary atherosclerosis in familial hypercholesterolaemia (FH). DESIGN: The effect of visceral fat accumulation on coronary lesions and risk factors in patients with FH was investigated. SUBJECTS: Thirty-one male patients with heterozygous FH who underwent coronary angiography. MEASUREMENTS: Abdominal fat distribution was estimated by a cross-sectional computed tomographic scan at the umbilical level. Plasma lipid, glucose and insulin concentrations and blood pressure were measured. A 75 g oral glucose tolerance test was also performed. RESULTS: The patients were divided into two groups according to the degree of visceral fat accumulation. Fifteen patients had high visceral fat accumulation (High VF group) and 16 patients had normal visceral fat accumulation (Normal VF group). Body mass index (BMI) and subcutaneous fat area were significantly higher in the high VF group. Baseline serum triglyceride was significantly higher and baseline low-density lipoprotein (LDL)-cholesterol and reduction of LDL-cholesterol during treatment were significantly lower in High VF group. Fasting plasma glucose and insulin concentrations, and systolic and diastolic pressures were significantly higher in the High VF group. Significant correlations were found between visceral fat area and the sum of the glucose and insulin concentration during an oral glucose tolerance test. Visceral fat area was significantly correlated with the severity of coronary stenosis index. Distal coronary lesions were significantly more frequent in the High VF group. Moreover, the correlation between the visceral fat area and coronary stenosis index was found to be independent of age, BMI, and subcutaneous fat area by multiple regression analysis. CONCLUSIONS: Visceral fat accumulation is a potent cardiovascular risk factor in heterozygous FH. PMID- 9226490 TI - Predicting intra-abdominal fatness from anthropometric measures: the influence of stature. AB - OBJECTIVE: To investigate the influence of height on the relationships between the intra-abdominal fat and anthropometric measures. SUBJECTS: Twenty healthy female volunteers aged 20-51 y from Aberdeen, and 71 men and 34 women aged 19-85 y from Nijmegen, The Netherlands. OUTCOME MEASURES: Intra-abdominal fat volumes by magnetic resonance imaging (MRI) in Aberdeen and cross-sectional areas at L4 L5 level by computerised tomography (CT) in Nijmegen, height, body mass index (BMI), waist circumference, waist sagittal and transverse diameters, waist to hip ratio, and skinfolds. RESULTS: In the MRI study the women with BMI 20-33 kg/m2, waist circumference 62-97 cm, height 148-172 cm, and intra-abdominal fat volume 0.07-2.66 kg, waist circumference gave the highest correlation of simple indices with intra-abdominal fat volume, explaining 77.8% of variance. Single cross sectional MRI cuts predicted volume with r = 0.94-0.99. Height in various levels of index power was not related to waist circumference, waist diameters, BMI, or skinfolds and did not improve prediction of intra-abdominal fat volume or of cross-sectional intra-abdominal fat area at any level. The CT study of men and women with BMI 18-32 kg/m2 and 19-38 kg/m2, waist circumference 71-112 cm and 74 125 cm, height 158-197 cm and 151-182 cm, and intra-abdominal fat area 13-274 cm2 and 19-221 cm2 respectively, height also had little influence on the relationships of intra-abdominal fat area with waist circumference or with any other indices of adiposity in linear or quadratic models. Compared to younger subjects, intra-abdominal fat area was higher in older subjects for a given waist circumference. CONCLUSIONS: Height does not importantly influence the differences in measures of adiposity or intra-abdominal fat volume in women, or intra abdominal fat area in both sexes. Age does influence the prediction of intra abdominal fat from waist circumference, but waist circumference alone has a predictable simple relationship with intra-abdominal fat volume or area, which is likely to relate to the prediction of health risk for health promotion. PMID- 9226491 TI - New age-adjusted measure of body fat distribution in children and adolescents: standardization of waist-hip ratio using multivariate analysis. AB - OBJECTIVE: To explore a new anthropometric index of body fat distribution adjusted for ages ranging from 6-15 y in both boys and girls. DESIGN: Sex, age, and 11 anthropometric variables were subjected to principal component analysis. Based on these results, we developed a new anthropometric index, namely an age adjusted measure of body-fat distribution. This index was evaluated statistically for suitability of use in epidemiological surveys. SUBJECTS: Japanese children, including obese and nonobese subjects, in one elementary and one junior high school in Yamanashi Prefecture, Japan: 508 boys and 549 girls whos ages ranged from 6 y 1 mon-15 y. MEASUREMENTS: Measurements included the height (Ht), body weight, circumference of the waist, hip and thigh. Body mass index, the ratios of the waist, hip or thigh to the Ht, waist-hip ratio (WHR) and waist-thigh ration were calculated. RESULTS: The first principal component (PC 1) accounted for 49.8% of the total variation, and was interpreted as an indicator of the general size on an individual. PC 2 accounted for 25.9%, and was interpreted as a shape measure that indicates body fat distribution. Calculation of WHR/Ht, a parameter that represented PC 2 adjusted by PC 1, gave an highly robust linear regression equation for age by gender. The residuals from the regression line for WHR/Ht deviated from normal distribution only in the boys, whereas the mean residual was nearly zero and distribution of the residuals was similar in three age subgroups by gender, supporting the use of the common standard deviation score in all age groups as an indicator of body fat distribution. CONCLUSION: The common standard deviation score of WHR/Ht can serve as an epidemiological index of body fat distribution adjusted for ages between 6 and 15 y. PMID- 9226492 TI - The prevalence of low back pain and associations with body fatness, fat distribution and height. AB - OBJECTIVES: To examine the associations of low back pain symptoms with waist circumference, height, waist to hip ratio and body mass index, and to test the interactions between (1) waist circumference and height, and (2) waist to hip ratio and body mass index. SETTING: Cross-sectional study set in The Netherlands of a random sample of 5887 men and 7018 women aged 20-60 y. RESULTS: The prevalences of low back pain in men and women in the past 12 months were 46% and 52%, of whom 17% and 21% had low back pain for a total of 12 or more weeks, and 13% and 18% had symptoms suggestive of intervertebral disc herniation. After adjustments for age, smoking and education, more women in the highest tertile of waist circumference reported low back pain in the past 12 months (odds ratio = 1.2, 95% confidence interval: 1.1-1.4), low back pain for a total of 12 or more weeks (odds ratio = 1.5, 95% confidence interval: 1.3-1.8), and intervertebral disc herniation symptoms (odds ratio = 1.3, 95% confidence interval: 1.1-1.6) than women in the lowest waist tertile. Corresponding values of low back pain symptoms for women with high body mass index or high waist to hip ratio were similar to those with high waist. There were no significant differences between men in different tertiles of waist, waist to hip ratio or body mass index reporting low back pain symptoms. Tallest subjects did not report low back pain symptoms more often than shortest subjects. There was no significant interactions between waist and height or between waist to hip ratio and body mass index on low back pain symptoms. CONCLUSIONS: Women who are overweight or with a large waist have a significantly increased likelihood of low back pain. There are no significant interactions between waist and height, or waist to hip ratio and body mass index on low back pain symptoms. PMID- 9226493 TI - Effect of food palatability on early (cephalic) phase of diet-induced thermogenesis in nonobese and obese man. AB - OBJECTIVE: To study the effect on the early (cephalic) phase of diet-induced thermogenesis (EDT) of palatable vs unpalatable food, in nonobese and obese man. SUBJECTS: Twenty-four nonobese volunteers and 19 obese clinic patients. DESIGN AND MEASUREMENTS: A palatable, liquid formula meal of Ensure (1048 KJ, 450 ml), and of Ensure made unpalatable by addition of aqueous KCl, were sipped on nonconsecutive mornings. O2 consumption (ml/min) was measured before, and starting 30, 60 and 90 min after beginning the test meal, from which EDT was calculated as KJ/min. RESULTS: Palatability of the test meal significantly increased EDT (palatability effect, P = 0.004) but obesity status per se, did not affect EDT. Nevertheless, the effect of palatability on EDT was dependent on obesity status, being seen only in the nonobese. EDT was significantly greater in the nonobese after the palatable than the unpalatable meal: (mean +/- s.e.m.) 2.45 +/- 0.14 vs 1.83 +/- 0.14; P < 0.0001, but not in the obese: 1.93 +/- 0.28 vs 1.73 +/- 0.20; P < 0.21. Therefore only after the palatable meal was EDT less in the obese compared with the nonobese: P < 0.05. The threshold for the unpleasant taste of added KCI was 31% higher in the obese than the nonobese: 4.2 +/- 0.4 vs 3.2 +/- 0.2 [g KCI]; P < 0.025. CONCLUSIONS: The early (cephalic) phase of dietary thermogenesis (EDT) is significantly increased in the nonobese by palatability, but not in the obese, so that only after a palatable meal is EDT less, or 'deficient,' in the obese compared with the nonobese. Also, the obese have a higher threshold for the unpleasant taste of KCI (in Ensure) than the nonobese. PMID- 9226494 TI - Insulin induces rapid changes of plasma leptin in lean but not in genetically obese (fa/fa) rats. AB - OBJECTIVES: To evaluate the plasma leptin concentration in lean and genetically obese fa/fa rats and to assess the response to 2 h hyperinsulinaemia. BACKGROUND: The recently discovered peptide leptin is a putative link between the size of the adipose mass and the hypothalamic centres controlling feeding behaviour. Several genetic models of animal obesity have been characterized as carriers of mutations of either the ob gene or leptin receptor. EXPERIMENTAL DESIGN: Lean (+/?) and obese (fa/fa) Zucker rats were studied under pentobarbital anaesthesia and underwent a 2 h euglycaemic hyperinsulinaemic clamp. Plasma leptin was measured in basal condition and at the end of the clamp study. Glucose rate of disappearance was evaluated by means of the isotope dilution technique using 3-3H glucose as tracer. RESULTS: fa/fa rats showed a 40 fold higher leptin concentration compared to lean littermates (0.47 +/- 0.10 vs 19.55 +/- 1.50 ng/ml, P < 0.0001). Euglycaemic hyperinsulinaemia increased plasma leptin in lean but not in genetically obese rats. CONCLUSIONS: Our results suggest that insulin may be a regulator of in vivo leptin secretion by adipose tissue of lean rates whereas it is ineffective in increasing plasma leptin in obese Zucker rats. PMID- 9226495 TI - Modernity, postmodernity and disability in developing countries. AB - This paper examines the implications of two theoretical perspectives, modernity and postmodernity, for provision of community-based disability services in developing countries. The author argues that modernity's embrace of the 'wonders' of science and technology have significantly affected our understanding of what community is. Modernity, in fact, leads us to view communities in one of two major ways: as inferior, or as ideal. Both views are deeply flawed. Postmodernity's profound scepticism of truth claims and authority provides a useful critique of community conceived in modern terms. The critique is helpful to the extent that it reveals the power of language in constructing our ideas of community. It also highlights a new way of thinking about participation, individualism and choice in community disability initiatives. PMID- 9226496 TI - The impact of a person's disability on his or her sibling. AB - The aim of this study is to determine the impact of a person's disability on the psychological, social, family and professional life of that person's siblings. The research is based on a clinical method and case studies. An analysis is made of siblings' verbalizations in different contexts and with different aims: unstructured research interviews conducted with persons who do or do not have a sibling with a disability; meetings of parents, professionals and siblings of persons with a disability; family discussion-therapy. The study demonstrates the importance of giving siblings the opportunity to express themselves, be it about their difficulty in dealing positively with their unique family situations in social and family contexts; about their feelings of shame and guilt; about the distress caused to them by their urges to identify with the sibling with a disability. If denied such opportunities, they may develop symptoms that can seriously compromise their futures. PMID- 9226497 TI - Rehabilitation efforts before and after tightening eligibility for disability benefits in Norway. AB - This paper reports the use of vocational rehabilitation among applicants for disability benefits in Norway before and after the eligibility criteria were tightened in 1991. The data sources were documents of 668 applicants from 1990 and 1993 in two countries. Vocational rehabilitation is the preferred benefit in the National Insurance Act, and was a core issue in the restriction reform. Nevertheless, rehabilitation was tried by only 14% of the applicants in 1990, and by no more than 19% in 1993. Eight per cent of the applicants were referred to an employment officer before determination in 1990, and 14% in 1993. In order to study different pathways from work to applying for disability benefits, six types of 'social security careers' were constructed. The commonest comprised sick pay only, the second led through unemployment, and the third included rehabilitation. Rehabilitation may also be offered to applicants who are refused disability benefits. This was the alternative determination for 4% of the refused cases in 1990, a proportion that increased to 12% after the reform. The main reason for the relatively infrequent use of rehabilitation is probably the continuous downgrading over many years of resources and institutions for this task. PMID- 9226498 TI - An international validation of the interaction with disabled persons scale. AB - The Interaction with Disabled Persons Scale (IDP) was devised to measure attitudes in terms of discomfort reported about social interaction with people with disabilities. The Scale has been used in Australia for ten years. This article reports results of an international validation project that was designed to determine whether psychometric characteristics and norms emerging for Australian groups apply elsewhere. A methodological proforma was developed to maximize uniformity of data collection across nine countries: Australia, Canada, Croatia, England, Germany, Hong Kong, Poland, Scotland and the United States. In most countries the Scale was administered in English; however it also was translated into Germany, Polish, French and Croat. Results indicate that across countries mean scores fell within ten points, similar moderate to high levels of item homogeneity occurred and level of prior contact with people with disabilities emerged as the strongest predictor of IDP scores. It was concluded that the IDP Scale is a valid measure that is able to discriminate between respondents within the countries included in the study. PMID- 9226499 TI - Sports participation in individuals with spinal cord injury: demographic and psychological correlates. AB - The factors associated with involvement in sport were surveyed in a sample of 121 adults (aged 16 to 60 years) with spinal cord injury. The subjects responded to a questionnaire requesting data on personal characteristics and injury variables, and completed measures of depression (Centre for Epidemiological Studies Depression Scale) and trait anxiety (Spielberger's State Trait Anxiety Inventory). Univariate analysis showed that although sport participants (n = 67) did not differ significantly from nonparticipants (n = 54) on any of the psychometric measures, they were younger, had sustained their injury at an earlier stage, reported higher incomes, and were less likely to have sustained cervical-level damage. A hierarchical discriminant function analysis revealed that the significant predictors discriminating sport participants from nonparticipants were age and income. These findings suggested that, at least for this sample of individuals with spinal cord injury, post-injury involvement in sport was not specifically associated with indices of psychological adjustment. PMID- 9226500 TI - Instrument for locating students with suspected learning disabilities: a quantitative approach. AB - The instrument to locate students with learning disabilities was developed to create equality and uniformity, so that such difficulties could be spotted independently of socialization factors, teachers and parents to whom the student had been exposed. The instrument was developed on the basis of research that defines the reading rate of students with reading disabilities in words per minute, the minimum number of errors for locating writing disabilities, and the number of answers and lines a student uses to reconstruct the content of a passage adapted to his or her age level. The research carried out in order to construct the instrument is the first to attempt to quantify disability, and on this basis to construct an instrument to locate students with disabilities in reading, writing and visual recall. The procedure can be carried out for a whole class in a single lesson period. One indirect conclusion from the research is that disability indicators remain with the learning disabled regardless of time since diagnosis or remedial help. Other implications related to combining the quantity component in diagnoses of degrees of difference in the disability identified, and in extra examination time for the learning disabled. The instrument should solve the problem of locating students with difficulty reading, writing or in visual recall, due to disability. Students thus identified will be directed to individual diagnosis to find out whether their difficulties are primary or secondary (e.g. a new immigrant who still has trouble with Hebrew). A personal corrective programme can then be constructed. PMID- 9226501 TI - Using the rehab scale in a Greek context. PMID- 9226502 TI - The transfer of technology towards the European assistive technology industry: current impediments and future opportunities. PMID- 9226503 TI - Technical aids for the physically handicapped: a psychological study of the master robot. PMID- 9226504 TI - Strength, balance and gait before and after kidney transplantation. PMID- 9226505 TI - Assessment of pressure relief characteristics in alternating pressure air cushions. PMID- 9226506 TI - A study of library based information needs of the parents of persons with mental retardation. PMID- 9226507 TI - Use of an acoustic orientation system by two adolescents with blindness and mental retardation: a 12-month follow-up. PMID- 9226509 TI - Pregnancy rates after intracytoplasmic sperm injection in relation to sperm recovery techniques. AB - PURPOSE/METHODS: Clinical outcome after intracytoplasmic sperm injection (ICSI) was evaluated in relation to three techniques of sperm recovery, mini-Percoll, simple concentration, and centrifugation and washing. RESULTS: Whereas fertilization and embryonic cleavage rates were similar in the three groups, the rates of implantation and clinical pregnancy were statistically significantly higher following sperm recovery by the techniques of mini-Percoll and centrifugation and washing. PMID- 9226508 TI - Genetic engineering: moral aspects and control of practice. PMID- 9226510 TI - Prognostic value of objective semen parameters in an in vitro fertilization program. AB - PURPOSE: The basic semen parameters seem to have a limited predictive value in male fertility. Could other objective sperm analyses be helpful in the choice of the most adapted assisted procreation technique? METHODS: This study concerns 78 infertile couples with insemination failures. For each semen, 21 objective parameters are analyzed in fresh semen and after sperm selection procedure. The 78 couples are then included in an IVF protocol and classified into two groups: fertile (at least one cleaved embryo is obtained) and infertile. RESULTS: Using multiple variant discriminant factorial analysis, we have found nine nonconventional parameters which induce us to define two classes of semen. These two classes fit with the classification into fertile and infertile groups in 74.4% of the cases. CONCLUSIONS: So these parameters allow us to predict the chance of obtaining embryos during an IVF trial and to choose for each couple the most appropriate technique: IVF or ICSI. PMID- 9226511 TI - Gonadotropins and glucocorticoid therapy for "low responders"--a controlled study. AB - PURPOSE: A randomized, nonplacebo controlled study was conducted to determine the effect of dexamethasone supplementation to a protocol of gonadotropin therapy in 42 "low-responder patients" aged 32 to 43 years. METHODS: All underwent at least two previous cycles treated by gonadotropins for unexplained infertility, or anovulation. Human menopausal gonadotropin was started on day 4 of the menstrual cycle combined with dexamethasone 0.5 mg administered nightly, as an adjuvant. A group of "low responders" who did not receive dexamethasone served as the controls. The number of follicles, total amount of gonadotropins used, time required for stimulation, fertilization, peak estradiol levels and pregnancy rate were evaluated. RESULTS: The number of developing follicles, estradiol levels, fertilization rate and pregnancy rate did not differ significantly. CONCLUSIONS: Although certain beneficial effects were observed in the literature in some of the infertile patients treated with corticosteroids, the overall results did not support daily, low-dose dexamethasone (long-acting corticosteroid) as a clinically useful adjuvant therapy for "low responders" during gonadotropin therapy. PMID- 9226512 TI - Analysis of the human zona pellucida during culture: correlation with diagnosis and the preovulatory hormonal environment. AB - PURPOSE: The objective of this study was to analyze sequentially the human zona pellucida changes in an in vitro fertilization program as it relates to several variables. METHODS: The zona pellucida thickness was measured daily in zygotes and cleavage-stage embryos on a Nikon inverted microscope equipped with Hoffman modulation contrast optics, using an ocular micrometer. A total of 512 embryos from 96 patients was evaluated. RESULTS: There was a highly significant direct correlation between zona thickness and preovulatory estradiol and basal day 3 FSH levels (P < 0.02 and P < 0.0006, respectively). This relationship showed a rapid reversal following 48 hr of culture; embryos from patients with the highest FSH levels had thinner zonae prior to transfer (P < 0.0007). The zonae from patients with unexplained infertility were thicker (19.4 +/- 2.7 microns) than those from patients with endometriosis (17.7 +/- 2.2 microns), tubal (17.5 +/- 2.4 microns), or male-factor infertility (16.4 +/- 2.7 microns) (P < 0.0001) on the first day of culture. CONCLUSIONS: We hypothesize that the thickness of the human zona pellucida is influenced by the preovulatory hormonal environment and diagnosis. These factors should be considered as part of the embryo quality evaluation prior to transfer or when assessing the possibility of using assisted hatching. More studies are needed to understand the factors regulating the thickness of the human zona pellucida. PMID- 9226514 TI - Late manifestation of pelvic abscess following oocyte retrieval, for in vitro fertilization, in patients with severe endometriosis and ovarian endometriomata. AB - PURPOSE: Our purpose was to study the unusual and rare late manifestation of severe pelvic abscess, following oocyte pickup (OPU), for in vitro fertilization and embryo transfer (IVF-ET). PATIENTS: The patients were three infertile women with stage IV endometriosis and ovarian endometriomata, as the sole reason for their infertility. Medical and surgical modalities to treat endometriosis and infertility proved to be unsuccessful. INTERVENTIONS: All patients were prepared for IVF-ET employing a long GnRH-a and hMG protocol. Transvaginal OPU was performed under ultrasound guidance. Intravenous (i.v.) prophylactic antibiotic was routinely administered. RESULTS: All women underwent ET, and one conceived. Forty, 24, and 22 days after OPU, respectively, these patients presented with acute symptoms of severe pelvic inflammatory disease (PID) and were found to have pelvic abscess. Broad-spectrum i.v. antibiotics were employed in all cases, however, two patients did not respond and bilateral adnexectomy was eventually performed. CONCLUSIONS: Severe endometriosis with ovarian endometriomata seems to be a significant risk factor for pelvic abscess development, following transvaginal OPU for IVF-ET. Prophylactic IV cefazolin does not seem to prevent this complication. Late manifestation of pelvic abscess supports the notion that the presence of old blood in an endometrioma provides a culture medium for bacteria to grow slowly after transvaginal inoculation. PMID- 9226513 TI - Treatment variables in relation to oocyte maturation: lessons from a clinical micromanipulation-assisted in vitro fertilization program. AB - OBJECTIVE: In an effort to understand the mechanism underlying the improved pregnancy rate observed in IVF cycles when gonadotropin-releasing hormone analogues (GnRH-a) are applied, we investigated a possible relationship between treatment variables and oocyte nuclear maturity. DESIGN: Nuclear maturity was retrospectively assessed in cumulus-free, denuded oocytes, obtained from women undergoing micromanipulation-assisted IVF treatment following controlled ovarian hyperstimulation with GnRH-a and menotropins. SETTING: The setting was the infertility and IVF unit of a tertiary academic medical center. PARTICIPANTS: Two hundred twenty-one patients underwent 435 treatment cycles. MAIN OUTCOME MEASURE: This was the proportion of germinal vesicle-intact immature (GVII) oocytes. RESULTS: One hundred fifty-four of the 3520 oocytes studied (4.4%) were in the GVII stage. These oocytes were found in 66 of the treatment cycles (15.2%) and in 54 of the patients (24.4%). Cycles in which GVII oocytes were detected did not differ from those in which all the aspirated oocytes were mature in the following respects: patient age, type and duration of infertility, controlled ovarian hyperstimulation protocol and time of ovum pickup. However, the GVII group was characterized by a significantly higher peak estradiol level, as well as a higher number of mature follicles visualized sonographically (diameter, > 14 mm) and oocytes retrieved. CONCLUSIONS: Comparing the present findings with previously published data, it appears that the inclusion of GnRH-a in the stimulation regimen is associated with a lower proportion of immature oocytes. A higher occurrence of oocyte-nuclear immaturity is apparently associated with a significantly better ovarian response to stimulation. The high incidence of immature oocytes observed in patients with normospermic partners and low fertilization rates in previous cycles may suggest that the fertilization failure in some of these cases is due to oocyte, rather than sperm, dysfunction. PMID- 9226515 TI - Is there really a decrease in sperm parameters among healthy young men? A survey of sperm donations during 15 years. AB - PURPOSE: Our purpose was to measure changes in semen quality and quantity in young healthy sperm donors in Jerusalem over time. METHODS: A retrospective analysis of semen parameters over 15 years using linear regression analysis, in a single sperm bank in a tertiary university center. Study population consisted of 188 young, healthy medical students, aged 20 to 30 years, who donated sperm samples for Artificial insemination between 1980 and 1995. RESULTS: There were no statistically significant changes in semen concentration and motility during the study period. The mean semen volume increased by 0.1 ml (5.1%) per year (P < 0.0001), with a concomitant mean rise of 5.8 x 10(6) (7.7%) per year in total motile sperm count. The percentage normal morphology decreased by a mean of 1.04% per year during the entire period (P < 0.0001). CONCLUSIONS: During the past 15 years, there has been an increase in total motile sperm count, secondary to an increase in semen volume, and a decline in normal morphology that are independent of the age and the duration of abstinence in fertile men. PMID- 9226516 TI - Reasons for rejecting potential donors from a sperm bank program. AB - PURPOSE: Recruiting donors to a sperm bank program is difficult and slow because of high dropout rates and high rejection rates. The profile of successful and unsuccessful donors was determined at our sperm bank. METHODS: A total of 199 men was screened from 1986 to 1994 in the anonymous sperm bank donor programs; 174 (87%) men dropped out or did not meet minimum guidelines. The study included 25 accepted donors and 20 rejected men (of 52 rejected donors, only 20 donors who came for two consecutive semen analyses were selected). Sperm quality variables and demographic data were compared between the groups. RESULTS: Accepted donors had significantly better semen quality in motility, velocity, linearity, and ALH than did rejected donors (P < 0.01). More rejected donors than accepted donors were single (P < 0.01). A higher percentage of accepted donors consumed caffeine (P < 0.001), and they were more likely to have college degrees (P < 0.03). CONCLUSIONS: These results indicate that loss of interest and poor semen quality were the major reasons for rejection of donors in our anonymous donor sperm bank program. PMID- 9226517 TI - In vitro maturation of human testicular sperm in patients with azoospermia. PMID- 9226518 TI - Response to Eisenberg and Schenker [The duties of ethical committees applied to human reproduction (J Assist Reprod Genet 1996;13:689-697)]. PMID- 9226519 TI - Response to Meldrum and Hamilton [monoamniotic twinning and zone manipulation: a survey of U.S. IVF centers correlating zona manipulation procedures and high-risk twinning frequency (J Assist Reprod Genet 1996;13-748)]. PMID- 9226520 TI - Intestinal transplantation. PMID- 9226521 TI - Megacystis-microcolon-intestinal hypoperistalsis syndrome. PMID- 9226522 TI - Recurrence of autoimmune hepatitis in children after liver transplantation. AB - BACKGROUND: Liver transplantation is recognized as the appropriate treatment for end-stage liver disease due to chronic active autoimmune hepatitis. While it was initially thought that the disease did not recur after transplant, it is now generally accepted that adult patients may develop recurrent disease, with studies reporting a recurrence rate of < or = 25%. We have noted a higher incidence of recurrent autoimmune hepatitis in our pediatric patients undergoing liver transplant, with a high incidence of associated morbidity. METHODS: We reviewed the records of six children followed up for autoimmune hepatitis who underwent orthotopic liver transplant for complications of end-stage liver disease. RESULTS: Of the six, five developed recurrent autoimmune hepatitis at a mean time of 11.4 months after transplant. The disease was aggressive, leading to cirrhosis and retransplant in three patients, within 1 year of recurrence. A second recurrence of disease occurred in all three retransplanted patients. One patient has received a third liver transplant, one has died, and one patient is asymptomatic on medical therapy. Autoimmune hepatitis recurred in all four patients receiving tacrolimus. CONCLUSION: We conclude that liver transplant for autoimmune hepatitis is likely to be palliative for most pediatric patients. Potent immunosuppressives such as tacrolimus do not protect against the development of recurrent autoimmune hepatitis. PMID- 9226523 TI - Colonoscopy or sigmoidoscopy as the initial evaluation of pediatric patients with colitis: a survey of physician behavior and a cost analysis. AB - BACKGROUND: Pediatric patients presenting with colitis, suggestive of inflammatory bowel disease, undergo evaluation with either flexible sigmoidoscopy or colonoscopy. Our objectives were to assess current practice behavior in the evaluation of pediatric patients with colitis and to determine whether flexible sigmoidoscopy or colonoscopy was more cost-effective as the initial evaluation. METHODS: Practice behavior and procedure charges were assessed using a nationwide survey, and costs for diagnostic strategies were compared using a decision analysis program. RESULTS: The vast majority of survey respondents would proceed with colonoscopy if colitis suggestive of Crohn's disease was noted in the rectosigmoid area (81%) or if ulcerative colitis extended proximal to the rectosigmoid area (70%). If colonoscopy would follow if flexible sigmoidoscopy suggested either ulcerative colitis or Crohn's disease (67%), then colonoscopy would result in a savings of 23%. If the evaluation was predetermined to be limited to flexible sigmoidoscopy (16%), then flexible sigmoidoscopy was the cost effective strategy with savings of 29%. If colonoscopy would follow flexible sigmoidoscopy for Crohn's colitis only (13%), there was no clear cost advantage. CONCLUSIONS: The most cost-effective strategy depends on the physician's need to know the disease location. Our survey results indicate that most physicians chose to establish the extent of disease in both ulcerative colitis and in Crohn's disease; thus initial colonoscopy would be the more cost-effective strategy. When knowledge of disease distribution is not essential for patient care, flexible sigmoidoscopy can lead to substantial cost savings. PMID- 9226524 TI - Can the histologic changes of cystic fibrosis-associated hepatobiliary disease be predicted by clinical criteria? AB - BACKGROUND: Correlation between clinical parameters and histology changes in cystic fibrosis liver disease has not been documented. The purpose of this study was to determine the histologic spectrum of cystic fibrosis liver disease and the degree to which a clinical scoring system can identify subjects with significant histologic abnormalities. METHODS: We reviewed the predictive value of physical examination, biochemical parameters, and a clinical liver score, incorporating physical examination and biochemical parameters, in predicting significant abnormalities of liver histology in 43 cystic fibrosis patients who underwent hepatic biopsy. Biopsies were scored by two masked pathologists for fibrosis, inflammation, inspissation, fatty infiltration, and congestion. RESULTS: Significant histologic disease was present in 56% of patients despite little biochemical or physical examination evidence of disease. No single parameter used in the scoring system predicted the type or degree of the liver disease. The clinical liver score had a sensitivity of 85% and a specificity of 82% in predicting significant histologic changes, yet it was unable to predict the specific lesion. CONCLUSIONS: Significant histologic liver disease is common in cystic fibrosis, although the exact nature of the lesion cannot be predicted without liver biopsy. A clinical liver score that was developed for this may be useful in determining which patients require more definitive evaluation. PMID- 9226525 TI - Nutrient accretion in preterm infants fed formula with different protein:energy ratios. AB - BACKGROUND: Although standard formulas for preterm infants promote intrauterine rates of weight gain, fat deposition in preterm infants fed these formulas has been reported to be considerably higher than that in the fetus. We hypothesized that a preterm infant formula with a higher protein:energy (P:E) ratio would promote accretion rates of fat, fat-free mass, and minerals closer to those of the fetus. METHODS: As part of a larger study to determine whether accretion rates of fat and fat-free mass closer to those of the fetus can be achieved with a higher P:E ratio, we present a descriptive analysis of 72-h nutrient balance studies performed on a subset (n = 15/30) of the infants randomly assigned to be fed formula with a P:E ratio of either 3.2 g/100 kcal or 2.6 g/100 kcal. RESULTS: Despite the higher intake and net absorption of nitrogen by infants fed the higher P:E formula, there was no statistically significant difference in net nitrogen retention between groups. There also were no statistically significant differences between groups in digestible energy, metabolizable energy, energy expenditure, or energy storage. Thus, partitioning of stored energy as protein and fat did not differ between groups. The retention of calcium, phosphorus, sodium, potassium, copper, and zinc also did not differ between groups, and nitrogen intake did not affect mineral retention. CONCLUSIONS: In this study, formula for preterm infants with a P:E ratio of 3.2 g/100 kcal vs. 2.6 g/100 kcal provided no apparent benefit in terms of the proportion of fat to lean tissue accretion as determined from nutrient balance data. PMID- 9226526 TI - Modified cow's milk formula with reduced renal acid load preventing incipient late metabolic acidosis in premature infants. AB - BACKGROUND: Premature infants receiving alimentation with cow's milk formulas are at a considerably high risk of developing incipient late metabolic acidosis, an early stage in the development of manifest late metabolic acidosis. Is it possible to reduce this risk by modification of the composition of a standard formula? METHODS: The mineral composition of a cow's milk preterm formula A was modified (formula B) with the aim of reducing the alimentary load to that of human milk. 160 premature infants were fed either mother's milk (n = 50) or the modified formula B (enriched with sodium and potassium) (n = 110), and their urine pH was tested twice a week. Randomly collected subgroups of infants were studied in detail for nutrient balances. The results were compared with earlier observations of 282 premature infants fed either mother's milk (n = 28) or the standard formula A (n = 254). RESULTS: Incipient late metabolic acidosis was observed in nine of 78 premature infants receiving mother's milk, 53 of 254 premature infants receiving the standard formula A, and only one of 110 premature infants fed the modified formula B. Net acid excretion was 0.58 mmol/kg/day in 11 premature infants receiving alimentation with the modified formula B compared with 1.73 mmol/kg/day in 23 premature infants fed formula A. This reduction was mainly due to an increased alkali excess (sodium + potassium-chloride) in intake and urine. CONCLUSIONS: Reduction of renal acid load with the modified formula B had a preventive effect on the rate of development of incipient late metabolic acidosis in premature infants. PMID- 9226527 TI - Treatment of childhood peptic esophagitis: a double-blind placebo-controlled trial of nizatidine. AB - BACKGROUND: Nizatidine is an H2 histaminic receptor blocker, which acts on the oxintic cells in the stomach. The efficacy of nizatidine on acid gastric secretion has been widely studied in adults with erosive and ulcerative esophagitis, but not in children. The aim of the present study was to evaluate the therapeutic efficacy of nizatidine in children with reflux esophagitis. METHODS: Twenty-six patients were studied; all of them underwent endoscopy with multiple esophageal biopsies and 24-h intraesophageal pH monitoring. The diagnosis of esophagitis was based on histologic features. Patients were randomly assigned to double-blind treatment with either nizatidine or a placebo (10 mg/kg/day in two doses) for 8 weeks. A symptomatic score assessment was evaluated during the study. RESULTS: Twenty-four patients completed the 8-week protocol. After therapy, 9/13 (69%) patients on nizatidine and 2/13 (15%) patients on the placebo were healed (p < 0.007 by Fisher's exact test). Histological findings were improved in two other (16.7%) patients and unchanged in the last (8.3%) patient on nizatidine. In the placebo group there was histological improvement in three (25%) patients, no variation in six (50%), and worsening in one (8.3%). After therapy, determination of esophageal pH showed a statistically significant decrease of the total acid exposure time (p < 0.01) only in the nizatidine group. The clinical score analysis showed an improvement of symptoms only in the nizatidine group (p < 0.01), except for vomiting, which was reduced in both groups. CONCLUSIONS: Our results show that nizatidine is effective in treating children with reflux esophagitis. The children included in this study did not have severe esophagitis, and the conclusion must be limited to those with mild to moderate degrees of disease. PMID- 9226528 TI - Prospective significance of antiendomysium antibody positivity in subsequently verified celiac disease. AB - BACKGROUND: In order to assess their long-term predictability for the diagnosis of celiac disease, antiendomysium antibody results were compared with the outcome of the Interlaken diagnostic process. METHODS: Prospective gluten challenge was performed in 153 children with previously diagnosed flat small-intestine mucosa. In 90 patients (Group A), endomysium antibodies were initially positive, in seven (Group B) they were negative, and 56 patients (Group C) had no initial serological results. In IgA-deficient persons, IgG antibodies were also assayed, both by the immunofluorescent method. RESULTS: Histological relapse rates were 100% (90/90), 14.3% (1/7), and 76.8% (43/56), p < 0.001, in Groups A, B, and C, respectively. Each patient with relapse also exhibited endomysium antibody positivity during the challenge. Patients in whom celiac disease could be finally ruled out remained consistently endomysium-antibody negative. The celiac disease patient in Group B had severe secondary immunoglobulin deficiency at entry, which explained the initial negativity. Diagnosis based on antiendomysium antibody positivity and flat mucosa gave a higher applicability (92.8 vs. 50.3%) and reliability (relapse rate 100 vs. 89.6%) than the 1990 European Society of Paediatric Gastroenterology and Nutrition (ESPGAN) criteria among these patients. CONCLUSIONS: Endomysium antibody positivity at presentation has been found to be as useful as gluten challenge in the diagnosis of celiac disease, even in patients under the age of 2 years. Challenge is still advisable in patients with a flat small intestinal mucosa when antiendomysium antibody results are negative or have not been done, as among these patients significantly lower relapse rates were found. PMID- 9226529 TI - Use of barbiturates in the treatment of cyclic vomiting during childhood. AB - BACKGROUND: Cyclic vomiting is an uncommon disorder that can be described as recurrent, self-limiting, fairly uniform episodes of intractable nausea and vomiting with no identifiable organic cause, separated by symptom-free intervals. There is no established therapeutic regimen for this disorder. METHODS: Fourteen children referred to the Pediatric Gastroenterology Clinic were diagnosed with cyclic vomiting from May 1984 to January 1995. Vomiting, the predominant symptom, was present in all children and was severe enough to require hospitalization in 11. Associated symptoms included abdominal ain, headache, nausea, aura, and fever. Diagnostic studies were done to rule out organic causes as indicated in individual patients. Daily phenobarbital was prescribed in all 14 patients. The dose ranged from 30 to 120 mg/hs, (mean 2 mg.kg-1.day-1), with a median dose of 60 mg/hs [corrected]. Prior therapy with propranolol (3 patients) and butalbital (2 patients) had been ineffective. RESULTS: Eleven patients had complete resolution of their symptoms, and 3 patients had marked improvement in their symptoms with infrequent attacks of reduced severity. The only side effects associated with long-term phenobarbital therapy were behavioral in nature, namely hyperactivity and disruptive behavior at school. CONCLUSIONS: The results of our series of 14 patients, all of whom received barbiturates, support the usefulness of this therapeutic approach. Hence we feel that daily low-dose phenobarbital therapy is a safe and effective therapy in preventing episodes of cyclic vomiting in children. PMID- 9226530 TI - Measurement of the rate of entry of intact colon-derived lactose into the circulation: a model for assessing gut uptake of molecules not endogenously synthesized. AB - BACKGROUND: Results of in vitro studies have documented colonic absorption of lactose in the newborn. A stable isotope model was developed for assessing the entry rate of intact lactose into the portal circulation in newborn piglets. METHODS: In experiment 1, unlabeled and [D-1-(13C)]-lactose were infused into two separate mesenteric veins, and in experiment 2, labeled lactose was infused into a mesenteric vein and unlabeled lactose was infused into the colon. The 13C enrichment of plasma lactose was assessed by high performance liquid chromatography gas chromatography combined with mass spectrometry. RESULTS: The isotopic estimate of the mesenteric venous infusion rate of lactose was 91% of the theoretical. In the second experiment 13% of the unlabeled lactose infused into the colon reached the portal circulation. CONCLUSIONS: The current study provides the first, direct, in vivo confirmation of colon absorption of intact lactose. The tracer model could be used to evaluate intestinal or colonic absorption of other organic compounds not endogenously synthesized, including vitamins or drugs. PMID- 9226531 TI - Normal gastric histology in Helicobacter pylori-infected children. AB - BACKGROUND: In adults, Helicobacter pylori infection is always associated with gastritis or ulcer. However, very active gastritis and ulcers are rarely seen in children. The aim of the present work was to study the relationships between H. pylori and gastric mucosa in children. METHODS: Eighty infected children and adolescents including 48 (60%) neurologically impaired institutionalized patients, aged 2 months-22 years (mean 11.7 +/- 5.2 years) were studied retrospectively. All the patients underwent gastroscopy, and three antral and two fundic biopsy specimens were taken for histology and bacteriology. RESULTS: A normal gastric mucosa was found in 22 of 80 patients (27.5%), whereas the others had gastritis (n = 58, 72.5%). There were no statistical differences between patients with normal histology and those presenting with gastritis for age, sex, ethnic background, symptoms, and the degree of bacterial colonization. The macroscopic aspect of gastritis was less frequently found in children with a normal histology compared with those with histological gastritis (p < 0.001). CONCLUSIONS: These data show that H. pylori infection can be associated with a normal gastric histology in children. PMID- 9226532 TI - Increased TIA1-expressing intraepithelial lymphocytes in cow's milk protein intolerance. AB - BACKGROUND: Although a high level of intraepithelial lymphocytes (IELs) has been demonstrated in intestinal biopsies from children with cow's milk protein intolerance (CMPI), the properties of IELs in food-sensitive enteropathies remain unclear. In the present study, we analyzed the cytotoxic potential of IELs in CMPI, using a monoclonal antibody directed against the cytotoxic granule associated protein TIA1. METHODS: The study included 18 duodenal biopsies from 10 children previously diagnosed with CMPI and on a cow's milk-free diet of various duration. Six normal duodenal biopsies from children free of food intolerance served as controls. Immunostaining of formalin-fixed tissues was used to determine in the intraepithelial compartment (1) the number of TIA1-expressing cells per 100 epithelial cells, (2) the number of IELs per 100 epithelial cells, (3) the ratio of TIA1-expressing IELs (TIA1/IEL ratio). RESULTS: In CMPI, the number of IELs and TIA1-positive cells, as well as the TIA1/IEL ratio was significantly increased compared with controls. Moreover, a negative correlation between the TIA1/IEL ratio and the duration of the diet was observed. CONCLUSIONS: These results suggest that the recruitment of IELs with cytotoxic potential is increased in CMPI, and that IEL-mediated cytotoxicity could be involved in the pathogenesis of the disease. PMID- 9226533 TI - Prevalence of Helicobacter pylori infection in Nicaraguan children with persistent diarrhea, diagnosed by the 13C-urea breath test. AB - BACKGROUND: The impairment of gastric acid barrier caused by Helicobacter pylori (H. pylori) at the onset of infection may predispose to small bowel bacterial overgrowth, which could contribute to persistent diarrhea. METHODS: Using the 13C urea breath test, we determined the prevalence of H. pylori infection in 123 Nicaraguan children from Tipitapa, aged 1 to 65 months, from a low socioeconomic background. RESULTS: The overall prevalence of H. pylori infection was 77.2% (95/123). The prevalence varied with age and was significantly (p < 0.001) higher in infants < or = 12 months than in children aged 13-65 months, 91% (57/63) as against 63% (38/60). H. pylori infection was present in 44 of 59 (75%) children suffering from persistent diarrhea compared with 51 of 64 (80%) age-matched asymptomatic controls. In the diarrheal group, 20 of 59 (34%) children presented with malnutrition, and 16 (80%) of them showed H. pylori infection. In the control group, 20 of 64 (31%) were malnourished, and 14 (70%) of them showed H. pylori infection. CONCLUSIONS: In Nicaragua, H. pylori is acquired in early infancy. The high prevalence among children in the first 12 months of life and the lower infection rate between 1 and 5 years of age suggest a loss or clearance of infection, also an occasional finding in adults. H. pylori infection appears to be not a risk factor for persistent diarrhea or malnutrition in Nicaraguan children. PMID- 9226534 TI - NADPH-diaphorase in antral gastritis of childhood. AB - BACKGROUND: Nitric oxide has an important role in the pathophysiology of the gastric mucosa. However, to date, it is not clear if nitric oxide plays a cytoprotective or cytotoxic role in the pathogenesis of mucosal lesion. METHODS: We have used the NADPH-diaphorase histochemistry that selectively stains cells containing nitric oxide synthase, the enzyme that catalyzes the production of nitric oxide on the antral gastric mucosa of children with antral gastritis. RESULTS: In the lamina propria of the mucosa, the presence of the enzymatic activity was found in perivascular round cells and nerve fibers. In the epithelium, focal positivity to NADPH-diaphorase was found in superficial cells, mainly located in the extrusive zones. The epithelial cells in the pits and glands were negative, in the mucous layer, Helicobacter pylori were also stained by NADPH-diaphorase. A single H. pylori-infected child who was also examined after eradication of the H. pylori showed during the control examination absence of microorganisms and reduction of the NADPH-diaphorase-positive cells in the mucosa. CONCLUSIONS: Our results demonstrate that, in gastric mucosa, endogenous and exogenous structures express a NADPH-diaphorase activity. PMID- 9226535 TI - Introduction of 6-mercaptopurine in Crohn's disease patients during the perioperative period: a preliminary evaluation of recurrence of disease. AB - BACKGROUND: Recurrence of Crohn's disease after surgery is a common occurrence, pointing to the need for a strategy to prevent recurrent disease. We report the postoperative course of 10 patients who required intestinal resections for complications related to Crohn's disease. METHODS: All patients had a Pediatric Crohn's Disease Activity Index score of 10 or greater. Among these patients, 5 began treatment with 6-mercaptopurine in the perioperative period. All 10 had received various combinations of prednisone and salicylate compounds. Patients who were given 6-mercaptopurine did not discontinue the medication until 2 years after the surgery. RESULTS: To date, none of the five patients who were placed on 6-mercaptopurine have had recurrence of their Crohn's disease (mean disease-free period 32.6 +/- 18.4 months). Among those five patients not receiving 6 mercaptopurine there have been three relapses (mean time to relapse 3.7 +/- 1.2 months). Log-rank sum analyses of Kaplan-Meier survival curves show benefit to patients receiving 6-mercaptopurine in preventing relapses after intestinal resection (p < 0.05). CONCLUSIONS: Although the underlying pathophysiologic reasons leading to the high relapse rate after intestinal surgery in Crohn's disease are unknown, we conclude that treatment with 6-mercaptopurine in the perioperative period may be warranted to help prevent the recurrence of Crohn's disease after surgery. PMID- 9226536 TI - Acute steatosis in an 8-year-old boy with insulin-dependent diabetes mellitus. PMID- 9226537 TI - Concurrence of celiac disease and juvenile dermatomyositis: result of a specific immunogenetic susceptibility? PMID- 9226538 TI - Balloon dilatation of a compromised splenorenal shunt. PMID- 9226539 TI - Pancreatitis in patients with inflammatory bowel disease. PMID- 9226540 TI - Method for assessing absorption. PMID- 9226541 TI - Preventing relapse after surgery for Crohn's disease: where do we go from here? PMID- 9226542 TI - Childhood intussusception: management perspective in 1995: what to do if it is recurrent. PMID- 9226543 TI - Childhood intussusception: management perspectives in 1995. PMID- 9226544 TI - Vitamin E supplementation in cystic fibrosis. PMID- 9226545 TI - Enzymatic hydrolysis of the conjugate of vitamin D and related compounds. AB - The monoglucuronides of vitamin D, 25-hydroxyvitamin D and the corresponding pro forms were subjected to enzymatic hydrolysis using beta-glucuronidase, and substrate specificities were found in the examined enzymes originating from different sources, which were determined using reversed-phase high-performance liquid chromatography with UV detection. The enzymatic hydrolysis of the corresponding monosulfates was also examined using the same system. PMID- 9226546 TI - Time resolved study of effect of chlorpromazine on mobility of cytochrome P-450 and phospholipids in the inner membrane of adrenocortical mitochondria. AB - The effects of chlorpromazine on the mobility of cytochrome P-450 and the fluidity of lipid membranes have been investigated in bovine adrenocortical submitochondrial particles (SMP). Rotational diffusion of the cytochrome was measured by observing the decay of absorption anisotropy, ra(t), after photolysis of the heme.CO complex by a vertically polarized laser flash. Analysis of ra(t) was based on a 'rotation-about-membrane-normal' model. The anisotropy decayed within 2 ms to a time independent value r3. The presence of chlorpromazine decreased the mobile population of cytochrome P-450 from 28 to 23%. The rotational relaxation time phi a of the mobile population (approximately 1100 microseconds) was, however, not significantly changed by chlorpromazine. The lipid fluidity was examined by observing time-resolved fluorescence anisotropy, rf(t), of 1,6-diphenyl 1,3,5-hexatriene (DPH). The anisotropy rf(t) decayed within 70 ns to a time independent value r infinity. The motion of DPH was analyzed based on a 'wobbling-in-cone' model. The presence of chlorpromazine decreased the cone angle from 42 degrees to 39 degrees, while the rotational relaxation time phi f (approximately 2 ns) was not significantly changed by the presence of chlorpromazine. These results demonstrate that chlorpromazine decreased the mobility of not only lipids but also membrane proteins. PMID- 9226547 TI - A sensitive and selective method for the determination of mevalonic acid in dog plasma by gas chromatography/negative ion chemical ionization-mass spectrometry. AB - A sensitive and selective method has been developed for the determination of mevalonic acid (MVA), a cholesterol biosynthetic precursor, in dog plasma using solid-phase extraction in combination with gas chromatography/negative ion chemical ionization-mass spectrometry (GC/NICI-MS). MVA extracted from plasma with a phenylboronic acid-bonded phase cartridge was converted to its pentafluorobenzyl (PFB) ester-cyclic boronate derivative to produce a carboxyltate anion [M-PFB]- in the NlCl mode. PFB ester boronate derivatives of MVA and its internal standard, d3-mevalonolactone, were monitored in the selected ion mode at m/z 213 and 216, respectively. The precision and accuracy of within run and between-run assays were within 8%. This method was used to follow the diurnal variation of MVA levels in plasma of fasted and fed dogs. The diurnal variations of plasma MVA levels observed between the two groups were similar to those reported previously for human and rat plasma. PMID- 9226548 TI - Digital fluorescence imaging of elementary steps of neurosteroid synthesis in rat brain glial cells. AB - With fluorescence microscopic imaging, we have examined Ca2+ signaling, LDL uptake and distribution of cytochrome P450 scc on individual rat brain glial cells in order to investigate the molecular mechanisms of neurosteroid synthesis. Astrocytes and oligodendrocytes were cultured from newborn rat brain. Ca2+ signaling was observed in Calcium Green-1 loaded astrocytes upon neurotransmitter stimulations using video-enhanced microscopy. Upon stimulation of serotonin and glutamate, we observed typically three types of Ca2+ signaling which were Ca2+ oscillations, a transient increase in Ca2+ concentration and Ca2+ oscillations superimposed on a transient Ca2+ increase. On the other hand, histamine and ATP induced only a transient increase in Ca2+ without oscillatory response. Uptake of octadecyl rhodamine (R18) labeled LDL by astrocytes and oligodendrocytes was observed in the time scale of 30 min with confocal laser scanning microscopy. Some localization of LDL in the cytoplasm was observed for astrocytes. For oligodendrocytes, incorporated LDL was distributed over the entire cytoplasmic region of both cell body and multiple branched cell processes. The presence of a significant amount of cytochrome P450 scc was demonstrated with immunofluorescence staining in both astrocytes and oligodendrocytes. The density of P450 scc in both glial cells was suggested to be around 1% of that in bovine adrenocortical fasciculata cells. The results lead to an improved quantitative picture of neurosteroid synthesis in glial cells. PMID- 9226549 TI - Chiral analysis of 3-methoxy-4-hydroxyphenylglycol in human urine. AB - Since 3-methoxy-4-hydroxyphenylglycol (MHPG) is a neutral metabolite from norepinephrine, it will be a diagnostic marker for mental diseases such as depression. For the development of an immunoassay, the natural enantiomer of MHPG would be required to prepare its antigen and to examine the specificity of the antibody. A natural enantiomer synthesized, however, has not been obtained so far. In this paper, we attempted to enantioseparate synthetic DL-MHPG and to assign D-enantiomer from the optical rotation of MHPG purified from human urine, because endogenous norepinephrine occurs as D-enantiomer which should metabolically generate D-MHPG. Enantioseparation conditions were tested using a Ceramospher Chiral RU-1 column (4.6 x 250 mm) at a flow rate of 0.5 ml/min. The resolution was adequate for the analysis and purification of synthetic DL- and the urinary MHPGs using methanol as a mobile phase and the column temperature at 0 degrees C, where DL-MHPG was detected as two peaks. The earlier peak (peak 1) showed (-) optical rotation, while the latter gave (+) optical rotation. After being treated with beta-glucuronidase, the normal human urine was extracted with ethyl acetate and then evaporated to dryness. The residue was suspended in water and the supernatant was analyzed and purified by a reversed phase column with a multi channel detector. A peak corresponding to MHPG was collected and concentrated to dryness. The pooled residues were dissolved in methanol and enantioseparated on the chiral HPLC. The urinary MHPG appeared as a single peak which was corresponded to the earlier peak of DL-MHPG and showing (-) optical rotation. Thus, the urinary MHPG was found to be D-(-)-MHPG. Then the absolute configuration of enantioseparated MHPGs were assigned to each optical rotation, judging from the chemical data and the metabolic pathway of the urinary D-MHPG. These enantiomers will be useful for studying on biochemistry and immunoassay. PMID- 9226550 TI - Development of enzyme immunoassay of 2'-deoxycytidine. AB - In order to study 2'-deoxycytidine (2'-dCyd) as a possible prognostic marker in cancer chemotherapy, an enzyme immunoassay (EIA) was developed. 2'-dCyd as a hapten was succinylated and two omicron-monosuccinyl-2'-dCyd's were purified by high performance liquid chromatography and identified by mass spectrometry and 1H NMR. Two antigens were prepared by conjugating two omicron-succinyl derivatives with keyhole limpet hemocyanin (KLH) as a carrier. Both antigen produced specific antibodies to 2'dCyd in BALD/c mice. The spleen cells of one mouse immunized with 5'-omicron-succinyl-2'-dCyd-KLH were hybridized with myeloma cells. One monoclone selected produced a specific antibody. A convenient EIA was attained by using the monoclonal antibody. PMID- 9226551 TI - Capillary electrophoretic analyses of beta-trace protein and other low molecular weight proteins in cerebrospinal fluid from patients with central nervous system diseases. AB - Ordinary capillary-zone electrophoresis (CZE), as well as CZE in a sodium dodecylsulfate-containing polymer solution (SDS-CZE) and capillary isoelectrofocusing (CIEF), was applied to the analysis of low molecular weight proteins in cerebrospinal fluid (CSF) from patients with various neuropsychiatric disorders. Under the CZE conditions employed, a peaks of beta-trace protein (beta TP), which is the most abundant low MW protein in CSF, was clearly detected on the electropherograms of all the samples examined, and the CSF beta TP level could be tentatively determined using allylamine added at a constant concentration as the internal standard. The results revealed that beta TP in CSF was non-specifically increased in organic disease in the central nervous system (CNS), especially in ones giving severe physical damage to the brain tissues. On the other hand, SDS-CZE allowed us to determine simultaneously the CSF minor low MW proteins other than beta TP, such as beta 2-microglobulin, gamma-trace protein, myelin basic protein, etc., while the CIEF electropherograms suggested that beta TP were separated into several fractions with the different PI values. These capillary electrophoresis systems seem to be powerful as aids in the biochemical examinations of beta TP and other low molecular weight proteins in CSF from patients with CNS diseases. PMID- 9226552 TI - Determination of cefuroxim levels in human serum by micellar electrokinetic capillary chromatography with direct sample injection. AB - Effective monitoring of cefuroxim levels by micellar electrokinetic capillary chromatography with direct serum injection are discussed and compared with the HPLC method. With capillary electrophoresis (CE), in contrast to HPLC, good resolution and efficiency was demonstrated as well as low consumption of solvent and samples. The CE system was applied at 15 kV with UV detection at 274 nm using 150 mM sodium dodecyl sulfate in 20 mM sodium phosphate and borate (pH 9.0) as electrolyte. The results were seen within 12 min with efficiency approaching 182,000 theoretical plates. The coefficients of variations of migration time and peak area were less than 0.8 and 5.9%, respectively. The detection limits for quantitative determination were 0.28 microM level. Good linearity and recovery were also obtained in the range of serum levels usually encountered in clinical analysis with a correlation coefficient of r = 0.991 and 98-101% recovery. The monitoring of cefuroxim in human serum with micellar electrokinetic capillary chromatography (MECC) is demonstrated. Identification of cefuroxim in human serum with MECC is demonstrated. Identification of cefuroxim was performed by characterizing the sample peak in terms of the migration time and UV spectrum. Considering the results of our study, the CE method should by highly suitable for the separation of cefuroxim in biofluids. PMID- 9226553 TI - Determination of amino acids in biological fluids by capillary gas chromatography with nitrogen-phosphorus selective detection. AB - A selective and sensitive method for the determination of protein and non-protein amino acids in biological fluids by capillary gas chromatography (GC) has been developed. The amino acids in the samples were directly converted into their N(O,S)-isobutoxycarbonyl methyl ester derivatives and measured by GC with nitrogen-phosphorus selective detection (NPD) using a DB-17ht capillary column. Using this method, the derivatives of the 21 protein amino acids and the 25 non protein amino acids provided excellent NPD responses and were quantitatively and reproducibly resolved within 28 min. The lower detection limits of these amino acids, at a signal-to-noise ratio of 3, were ca. 6-150 pg injected. The calibration curves for each amino acid in the range of 0.02-2 micrograms were linear and sufficiently reproducible for quantitative analysis. This method was successfully applied to small urine and serum samples without prior clean-up; there was no evidence of interference from coexisting substances. Overall recoveries of amino acids added to urine and serum samples were 83-112%. The intra-assay and inter-assay R.S.D. of amino acids in these samples were 0.3-8.9% (n = 3) and 1.9-15.8% (n = 3), respectively. PMID- 9226554 TI - The influence of temperature on the multiple separation of estrogenic steroids using mobile phases modified with beta-cyclodextrin in high-performance liquid chromatography. AB - The effect of temperature on the retention and multiple separation of six estrogenic steroids in reversed-phase liquid chromatography has been studied. Capacity factors (k') of estriol, 17 beta-estradiol, 17 alpha-estradiol, d equilenin, equilin and estrone were measured using mobile phase modified with different concentrations of beta-cyclodextrin (from 0-16 mM), a fixed solvent composition (acetonitrile-water) and a wide range of column temperatures (from 5 to 80 degrees C). The plots of capacity factors vs. reciprocal of absolute temperature are nonlinear in each case when mobile phase modified with beta cyclodextrin was used. Particularly strong nonlinearity was observed at lower temperature and at higher beta-cyclodextrin concentration. The complex chromatograms were evaluated using optimization parameters such as capacity factor of the last-eluted peak (k'max), the smallest resolution between adjacent peaks (Rs,min) and relative resolution product (r). The results presented describe precisely the role of temperature in high-performance liquid chromatography systems in which mobile phases modified with cyclodextrin were used. Moreover, the elution order of estrogenic steroids on modified and unmodified mobile phases has been discussed. PMID- 9226555 TI - Analysis of urinary and faecal porphyrin excretion patterns in human porphyrias by fast atom bombardment mass spectrometry. AB - We report a new method for obtaining urinary and faecal porphyrin excretion patterns in human porphyrias based on fast atom bombardment mass spectrometry (FAB-MS). Porphyrins were esterified and extracted from urine or faeces as their methyl esters for analysis by FAB-MS. The protonated pseudo-molecular ion [M + H]+ observed for each porphyrin is characteristic of that porphyrin, thus allowing a mixture of porphyrins to be analysed without the need for chromatographic separation. By using tandem MS, identification and characterisation of unknown porphyrins can be achieved. The urinary and faecal porphyrin excretion patterns from various porphyric patients obtained by FAB-MS are in good agreement with those analysed by TLC or HPLC methods. PMID- 9226556 TI - Charge-transfer chromatographic study of the interaction of antibiotics with sodium dodecylsulfate. AB - The interaction of 29 antibiotics with the anionic surfactant sodium dodecylsulfate (SDS) was studied by charge-transfer reversed-phase chromatography carried out on impregnated silica layers using water-methanol mixtures as eluents. The hydrophobicity of antibiotics and the relative strength of SDS antibiotic interaction was calculated separately for each antibiotic-SDS pair. SDS interacted with 17 antibiotics where the antibiotic-SDS complex was either more hydrophilic or more hydrophobic than the uncomplexed molecule. The relative strength of interaction depended considerably on the molecular structure of the antibiotics. No significant linear correlation was found between the hydrophobicity parameters of antibiotics and their capacity to interact with SDS. Stepwise regression analysis proved that the inductive effect of substituents, their electron-withdrawing power and proton-acceptor capacity exert a significance influence on the strength of interaction. PMID- 9226557 TI - Separation of oligomers of nonylphenyl ethylene oxide [correction of nonlphenylethylene oxide] on a porous graphitized carbon column. AB - The retention of nonylphenyl ethylene oxide oligomer surfactants was determined on a porous graphitized carbon (PGC) column using water--methanol mixtures as eluents. Linear correlations were calculated between the logarithm of the capacity factor (k') and the methanol concentration in the eluent. To test the validity of the hypothesis that in the case of homologous series of solutes the intercept and slope values are intercorrelated linear correlation was calculated between the two chromatographic parameters. To elucidate the role of the length of the polar ethylene oxide chain in the retention linear correlations were calculated between the chromatographic parameters and the number of ethylene oxide groups per molecule. Nonylphenyl ethylene oxide oligomers were well separated on the PGC column. Significant linear relationships were found between the corresponding chromatographic parameters indicating that the solutes behave as a homologous series of compounds. The retention of surfactants increased linearly with increasing number of ethylene oxide groups per molecule indicating hydrophilic interactions between the solutes and the surface of PGC support. PMID- 9226558 TI - Rapid gas chromatographic profiling and screening of biologically active amines. AB - An efficient method is described for the simultaneous determination of 57 amines including volatile aliphatic amines, nonvolatile polyamines and catecholamines present in aqueous samples. The method is based on two-phase isobutyloxycarbonylation (isoBOC) with a pH shift. In 1.0 M phosphate buffer at pH 7.5, phenolic hydroxyl groups were allowed to react with isobutyl chloroformate in the dichloromethane phase, and subsequently pH of the aqueous phase was increased to 12.0 for the reaction of basic amino functions. The resulting N(O)-isoBOC amines were recovered by solid-phase extraction using Chromosorb P in normal phase partition mode, with subsequent tert.- butyldimethylsilylation of the remaining hydroxyl groups for gas chromatographic analysis. Using this combined procedure, linear responses were obtained in the concentration range of 0.2-12 ppm, with correlation coefficients varying from 0.945 to 0.999 for most of the amines studied except for 5-methoxytryptamine (0.864). Temperature-programmed retention index (I) sets as measured on DB-5 and DB-17 dual-capillary columns of different polarity were characteristic of each amine and thus, useful in the screening for amines by computer I matching. When applied to saliva samples, the present method allowed rapid screening for each spiked amine and unspiked polyamines such as 1,3-diaminopropane, putrescine, cadaverine and spermidine. PMID- 9226559 TI - Raman spectroscopic analysis of isomers of biliverdin dimethyl ester. AB - The constitutional isomers of biliverdin dimethyl ester, IX alpha and XIII alpha, were studied by resonance Raman spectroscopy. The far-reaching spectral similarities suggest that despite the different substitution patterns, the compositions of the normal modes are closely related. This conclusion does not hold only for the parent state (ZZZ, sss configuration) but also for the configurational isomers which were obtained upon double-bond photoisomerization. Based on a comparison of the resonance Raman spectra, a EZZ configuration is proposed for one of the two photoisomers of biliverdin dimethyl ester IX alpha, while a ZZE, ssa configuration has been assigned previously to the second isomer. PMID- 9226560 TI - Keratin immobilized on silica as a new stationary phase for chromatographic modelling of skin permeation. AB - Skin permeability of organic compounds depends on their lipophilicity but can also be affected by compounds interactions with specific skin components. A good chromatographic model of percutaneous penetration determined solely by lipophilicity is provided by the immobilized artificial membrane (IAM) columns. To complete the model a new high-performance liquid chromatographic (HPLC) stationary phase was prepared by physical immobilization of keratin on silica support. The keratin immobilized on silica has properties typical for the reversed-phase materials but it retains specifically acidic solutes. The keratin column can be used to conveniently compare keratolytic properties of xenobiotics. It was demonstrated that retention parameters determined on a keratin column can be combined with the retention parameters determined on the IAM column to predict differences in skin permeability within a class of drugs. It has been postulated that HPLC can model skin permeation thus reducing research time and costs as well as the use of laboratory animals. PMID- 9226561 TI - Identification and mechanism of formation of potentially genotoxic metabolites of tamoxifen: study by LC-MS/MS. AB - On-line high-performance liquid chromatography-electrospray ionization mass spectrometry (HPLC-ESI MS) and tandem mass spectrometry (MS/MS) have been applied to the study of tamoxifen metabolism in liver microsomes and to the identification of potentially genotoxic metabolites. The results showed that the hydroxylated derivatives, including 4-hydroxytamoxifen and alpha-hydroxytamoxifen are detoxication metabolites, while arene oxides, their free radical precursors or metabolic intermediates, are the most probable species involved in DNA-adduct formation. PMID- 9226562 TI - Estimation of impurity profiles of drugs and related materials. Part 16: Identification of the side-products of the ethinylation step in the synthesis of contraceptive gestogens. AB - A new apolar impurity (3,17 alpha-diethinyl-13-ethyl-3,5-gonadiene-17-ol, IIb) was detected and identified in norgestrel with the aid of thin-layer and high performance chromatography and spectroscopic techniques. IIb is the product of the acid-catalysed dehydration of an overethinylated side product (Ib) of the ethinylation step in the synthesis of norgestrel. IIb can be determined by thin layer densitometry and high-performance liquid chromatography. Another impurity (17 alpha-ethinyl-13-ethyl-4-gonene-17-ol, IV), originating from a side product of the Birch reduction step in the synthesis of norgestrel was also detected and identified. The spot of IV overlaps with that of IIb in the TLC system of USP XXIII but can be separated and quantification by more selective TLC systems and by gas chromatography. PMID- 9226563 TI - Capillary electrochromatographic separation of amino acid enantiomers using on column prepared molecularly imprinted polymer. AB - The use of molecularly imprinted polymer polymerized in capillary for the separation of amino acid enantiomers by electrochromatography is described. The substrate-selective polymers were prepared by using L-phenylalanine anilide as print molecule and methacrylic acid and/or 2-vinylpyridine as the functional monomers, which is believed to interact both ionically and through hydrogen bonding with the print molecule. Several aspects of the polymer preparation were investigated, including the treatment of the inside surface of the capillary, the composition of the polymers and the running conditions of the capillary electrochromatography. Such separation was highly specific and depended on the presence of both the print molecule and the functional monomer in the polymerization mixture. This preliminary report demonstrates a novel and simple method for the development of the capillary electrochromatographic separation of amino acid enantiomers using molecularly imprinted polymer. PMID- 9226564 TI - Simultaneous determination of methylparaben, propylparaben and thimerosal by high performance liquid chromatography and electrochemical detection. AB - A reversed-phase high-performance liquid chromatographic method using amperometric detection has been developed for the analysis of methylparaben, propylparaben and thimerosal. The liquid chromatographic separation of the three preservatives was made possible on a C18-bonded silica column with a mixed solvent consisting of methanol and aqueous 0.02 M phosphoric acid (59:41, v/v). A potential value of +1.25 V versus Ag/AgCl was chosen for simultaneous analysis. The limits of detection were 1, 2 and 5 ng for a 20 microliters injection volume of methylparaben, propylparaben and thimerosal, respectively. The analysis time of less than 20 min in this study was found to be applicable for routine analysis of these compounds in pharmaceutical products. PMID- 9226565 TI - Enantioselective determination of oxprenolol in human plasma using dialysis coupled on-line to reversed-phase chiral liquid chromatography. AB - A fully automated method for the determination of the enantiomers of oxprenolol in human plasma was developed, involving dialysis through a cellulose acetate membrane, clean-up and enrichment of the dialysate on a short precolumn and subsequent chiral liquid chromatographic (LC) analysis. All sample handling operations were executed automatically by a sample processor equipped with a robotic arm (ASTED system). The trace enrichment column (TEC) was packed with octadecylsilica. After conditioning of the TEC with the LC mobile phase and pH 3.0 acetate buffer. After the enrichment step, the analyte was transferred by the LC mobile phase to the analytical column by means of a switching valve. The influence of different parameters of the dialysis process on the recovery of oxprenolol was first investigated using achiral LC conditions. The volume as well as the aspirating and dispensing flow rates of the acceptor solution were the main parameters studied. Oxprenolol was separated on a C18 stationary phase used for the enantioseparation of oxprenolol was a Chiralcel OD-R column which contained cellulose tris (3,5-dimethylphenylcarbamate) coated on silica as chiral selector. The corresponding mobile phase consisted of a mixture of pH 6.0 phosphate buffer containing NaClO4 at 0.45 M concentration and acetonitrile (70:30 v/v). UV detection was performed at 273 nm. The method developed was validated. Recoveries for each enantiomer of oxprenolol were about 80%. The method was found to be linear in the 50-2500 ng ml-1 concentration range (r2 = 0.999 for both enantiomers) and good results with respect to intra- and inter-day reproducibility as well as accuracy were obtained. PMID- 9226566 TI - Analysis of stereoisomeric C27-bile acids by high performance liquid chromatography with fluorescence detection. AB - A method for differentially measuring the 24-hydroxylated stereoisomeric intermediates (3 alpha,7 alpha,12 alpha,24-tetrahydroxy- and 3 alpha,7 alpha,24 trihydroxy-5 beta-cholestan-26-oic acids) and related C27-bile acids in beta oxidation of bile acid biosynthesis has been developed by high performance liquid chromatography with fluorescence detection. The method involved the derivatization of the above intermediable C27-bile acids into fluorescent esters with 3-(4-bromomethylphenyl)-7-diethylaminocoumarin, a newly synthesized labeling reagent for carboxylic acids. The fluorescent derivatives were subjected to a short silica gel column to eliminate interfering products prior to analysis by high performance liquid chromatography. The separation of the 16 kinds of bile acids containing stereoisomers was carried out using a reversed-phase Inertsil C8 column by gradient elution and detected with a fluorometer (Ex. 400 nm, Em. 475 nm). The linearity of calibration curve for each bile acid was from 0.5 to 250 pmol (r = 0.999) and the detection limits were about 15 fmol at a signal-to-noise ratio of 3. The method was applied to the determination of intermediates in beta oxidation of bile acid biosynthesis using rat liver homogenate. The results showed that two stereoisomers of 24-hydroxylated C27-bile acids were predominantly produced, indicating the formation of the isomers by the cis hydration with water. PMID- 9226567 TI - Optimization of a polarized photometric detector equipped with a split-type flow cell and its analytical application to oligo-saccharides. AB - A novel, non-modulated polarimeter called a polarized photometric detector (PPD) was previously described by the authors. The PPD enables the measurement of the optical rotation of chiral compounds as a change in absorbance by placing two linear polarizers on either side of a flow cell of a conventional photometric detector. The present study describes the optimization of the conditions of PPD for highly sensitive detection of saccharides. To maximize the light intensity, the light balancing filter and slit were removed from the detector (Shimadzu model SPD-10AV). These modifications resulted in an approximately 15-fold increase in the incident light intensity when the maximum current was applied to the lamp. When this intense light was transmitted through the polarizers, the signal intensity followed the theoretical equation for phase angles up to around 1 rad. If the energy of the transmitted light was less than 700 mV, however, the baseline noise was too great to determine the chiral analyte accurately. Setting the phase angle between two polarizers at 50 degrees and the detection wavelength at 400 nm provided the most suitable conditions. This detector was applicable for the determinations of oligosaccharides in foodstuffs separated by HPLC using gradient elution. PMID- 9226568 TI - A new small particle packing for faster analysis with high resolution. AB - The need for fast and efficient separations of complex samples such as pharmaceuticals and biologicals led to the development of fast, efficient, and reproducible 3.5 microns columns. Separations using 3.5 microns column are 30-50% faster at equal plate-count compared to 5 microns columns. The results show that the 3.5 microns columns (100 mm length) give the same efficiency and resolution of drug impurities as the 5 microns columns (150 mm length). For many analytical methods, switching to 3.5 microns columns saves time and reduces costs. Separation methodologies using 5 microns columns are easily modified to accommodate 3.5 microns columns of the same chemistry because efficiency, resolution and sensitivity remain the same. It is shown that 3.5 microns columns have lifetimes comparable to 5 microns columns of the same brand. PMID- 9226569 TI - Synthesis of a new tetradentate beta-diketonate-europium chelate and its application for time-resolved fluorimetry of albumin. AB - A new tetradentate beta-diketone chelator, 1,10-bis(8'-chlorosulfo dibenzothiophene-2'-yl)-4,4,5,5,6,6,7,7, -octafluorodecane-1,3,8,10-tetraone (BCOT), was synthesized. This compound forms a stable chelate with europium (III) and can be used as a fluorescence label for time-resolved fluorescence spectrometry. BCOT can be covalently bound to a protein under relatively mild conditions and, when complexed with europium(III), both in 0.1 M Tris-HCl solution and 1.0 x 10(-5) M tri-n-octyl phosphine oxide (TOPO)-0.05% sodium n dodecyl sulfate (SDS)-0.1 M NaHCO3 solution, forms a strongly fluorescent chelate having lifetimes of 225 microseconds and 240 microseconds, respectively. As a model reaction, bovine serum albumin (BSA) was labeled with BCOT in a carbonate buffer solution. Fluorescence properties of the labeled BSA solution were studied. A time-resolved fluorescence determination of the BSA(BCOT)40-Eu3+ solution was carried out. Detection limits of 9.3 x 10(-14) M (TOPO-SDS-NaHCO3 solution) and 6.7 x 10(-13) M (Tris-HCl solution) for BSA were obtained by the method. PMID- 9226570 TI - Ectodomain interactions of leukocyte integrins and pro-inflammatory GPI-linked membrane proteins. AB - Although glycosylphosphatidyl-inositol (GPI) linked membrane proteins do not possess transmembrane or cytosolic sequences they elicit transmembrane signals. Using microscopic fluorescence imaging and resonance energy transfer (RET) techniques we have shown that certain pro-inflammatory GPI-linked membrane proteins can interact with leukocyte beta 2 integrins (complement receptor type 3 (CR3) and 4 (CR4) and the leukocyte function-associated antigen-1 (LFA-1)). For example, physical associations between CR3 and Fc gamma RIIIB, CR3 and urokinase receptors, and CR3 and CD14 (lipopolysaccharide receptor) have been found. Although Fc gamma RIIIB appears to be constitutively associated with CR3, urokinase receptors and CD14 associations with CR3 are influenced by their ligation status and cell function (e.g. adherence and locomotion). CR3-to urokinase receptor interactions have been confirmed by immunoprecipitation techniques. Immunoprecipitation of CR3 from Brij-58 lysates after biotinylation of neutrophil membranes revealed proteins of M(r) = 40,000, 50,000, 74,000 and 120,000, in addition to bands corresponding to the integrin alpha and beta chains. Cell functions such as transmembrane signaling and superoxide release/priming have been linked to these interactions. Importantly, reagents that affect the lectin-like site of CR3, such as N-acetyl-D-glucosamine, alpha methyl-D-mannoside and beta-glucan alter these interactions and, in parallel, leukocyte functions. Thus, the interactions of GPI-linked proteins and integrins can be highly dynamic events linked to cell activities. Our studies suggest that it may be possible to develop new drugs directed at the lectin-like site of beta 2 integrins that block GPI-linked protein-to-integrin coupling thereby controlling inflammatory cell processes including cell adherence, locomotion and activation. Such drugs may be useful in clinical conditions such as ischemia reperfusion injury, sepsis, arthritis and others. PMID- 9226572 TI - Determination of a substance P antagonist in human plasma and urine using high performance liquid chromatography with ultraviolet absorbance and tandem mass spectrometric detection. AB - A high-performance liquid chromatographic (HPLC) assay using ultraviolet (UV) detection was developed and compared with a HPLC method with tandem mass spectrometric (HPLC/MS-MS) detection for the determination of a substance P receptor antagonist 2(S)-((3,5-bis(trifluoromethyl)benzyl)-oxy)-3(S)-phenyl-4-((3 oxo-1,2,4- triazol-5-yl) methyl)morpholine (Fig. 1, Ia, L-742 694) in human plasma and urine. The drug was isolated from the biological matrix through liquid liquid extraction. In the HPLC/UV method, the samples were initially injected onto a cyano Hypersil column, and the chromatographic region containing the peaks of interest was heart-cut onto an analytical C-18 Hypersil column via a column switching device. The analyte was quantified by monitoring absorbance at 205 nm. The limit of quantification for I extracted from 1 ml of plasma or urine was 2.5 ng ml-1, and the assays were validated in the concentration range 2.5-500 ng ml 1. The HPLC/MS-MS method were validated in the concentration range 0.2-500 ng ml 1. Both assays provided data with precision, measured as coefficient of variation, better than 10% at all points within the standard curve range and with adequate accuracy. PMID- 9226571 TI - Flow injection analysis for measurement of activity of matrix metalloproteinase-7 (MMP-7). AB - A simple and convenient method for measuring the activity of a recombinant human matrix metalloproteinase 7 (MMP-7, matrilysin) was developed by flow injection analysis (FIA). For this method, purified recombinant MMP-7 zymogen expressed in E. coli and the substrate peptide (MOCAc-Pro-Leu-Gly-Leu-A2pr(DNP)-Ala-Arg-NH2) were used. Following the incubation of substrate peptide with activated r-proMMP 7, the resulting fluorescent product peptide (MOCAc-Pro-Leu-GLY) was monitored with a fluorescence detector (lambda ex 328 mm, lambda em 393 mm) without chromatographic separation. In this FIA system, the analysis time is 2 min and the standard curve is linear from 5 to 100 pmol of the product peptide injected. In order to use this FIA system as a method for screening inhibitors against MMP 7, the effects of CaCl2, EDTA and of the tissue inhibitor of metalloproteinase-1, and -2, were tested. A synthetic PRCGXPD-containing peptide (BS-10) was also observed to inhibit MMP-7 activity, with an IC50 value of 104 microM. Thus, it was concluded that the activity of r-MMP-7 can be reliably measured by the proposed system. Furthermore, to confirm the utility of this FIA system as a screening method, the inhibitory activity of the MMP-related substance in Joro spider (Nephilia clavata) venom was measured by this method. This inhibitory activity was observed in an extract of a venom diluted 1000-fold. Thus, the FIA method is not only simple and quick, but also sensitive enough to screen and analyze the inhibitory properties of a large number of test compounds. PMID- 9226573 TI - Determination of homocysteine and other thiols in human plasma by capillary electrophoresis. AB - A new capillary electrophoresis (CE) assay for thiols in human plasma, including homocysteine, which is an indicator of several clinical states has been developed. The thiols were derivatized quantitatively at 50 degrees C, pH 8.0 with a fluorogenic reagent, ADB-F (4-aminosulfonyl-7-fluoro-2,1,3 benzoxadiazole), which is about 30 times faster compared to the other fluorogenic reagent, SBD-F (ammonium 7-fluoro-2,1,3-benzoxadiazole-4-sulfonate). The separation of ABD-thiols was performed in a 50 mM sodium phosphate buffer (pH 2.1) using a bare fused silica capillary (27 cm x 50 microns i.d.) at 25 degrees C. With the electric field of 560 V cm-1, the time needed for the separation of homocysteine, glutathione and cysteine was less than 8 min. A filter-type ultraviolet detector and a 512-channel diode-array detector (DAD) were employed for ABD-thiol analysis. DAD was used to confirm the ABD-thiol peaks. The limits of detection (S/N = 3) for homocysteine, glutathione, and cysteine were 0.5, 1 and 2 microM at 220 nm, respectively. PMID- 9226574 TI - Micromachined analytical devices: microchips for semen testing. AB - Micromachined devices (microchips) have been designed and tested for a range of clinically important assays. In this study we compare sperm motility determined using disposable glass microchips and a conventional Makler chamber. The 17 x 14 mm glass microchips contained three etched test structures each comprising either duplicate or quadruplicate analytical microchannels. Semen samples with sperm counts ranging from 21 to 78 million sperm per ml and forward progression scores of from 1+ to 3+ were evaluated and swimming times ranging from 360 s (3.3+ progression) to 770 s (1+,2 forward progression) observed in the microchips. Motility determined by the time taken for sperm to swim to the end of a microchannel (100 microns wide x 40 microns deep x 10 mm long) in the microchip correlated with forward progression of the sperm determined by the conventional Makler chamber method. This study demonstrates the feasibility of microchips for sperm motility testing and suggests that this technique would be applicable to the study of other types of motile cells. PMID- 9226575 TI - Determination of a novel anti-psychotic agent AD-5423 and its metabolites in plasma by high-performance liquid chromatography with fluorescence detection. AB - AD-5423, 2-(4-ethyl-1-piperazinyl)-4-(4-fluorophenyl)-5,6,7,8,9,10-hexahydrocy cloocta [b]pyridine, is a novel anti-psychotic agent. In order to investigate the pharmacokinetics of AD-5423, a simple and sensitive high-performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of AD-5423 and its two N-oxidized metabolites (N-desethyl AD-5423 and AD-5423 N-oxide) in plasma. After pretreatment of a plasma sample by solid phase extraction, AD-5423 and its metabolites were analyzed on a HPLC with fluorescence detection (335/410 nm). Chromatography was performed on two C18 reversed-phase columns connected by a switching system, with a mobile phase of acetonitrile-methanol-25 mM sodium phosphate buffer (pH 2.5) containing 25 mM sodium 1-heptanesulfonate (36:26:38 v/v/v). The method gives satisfactory accuracy and precision for the determination of AD-5423 and its metabolites. In human plasma, accurate determination are possible over the concentration ranges of 0.04-5 ng ml-1 for AD-5423 and 0.1-5 ng ml-1 for N-oxidized metabolites. The intra- and inter-day assay precision (R.S.D.) of AD-5423 (0.5 ng ml-1) were 3.6 and 7.2%, respectively. In plasma of experimental animals, the validated quantitative range are 0.1-100 ng ml-1 for both AD-5423 and its metabolites. PMID- 9226576 TI - Chromatography of crotamiton and its application to the determination of active ingredients in ointments. AB - Crotamiton, which is a mixture of cis and trans isomers, was investigated by several separation techniques. One of the HPLC modes, in which crotamiton eluted as a single peak, was selected for the determination of five active ingredients (crotamiton, prednisolone, glycyrrhetinic acid, dibucaine and chlorhexidine hydrochloride) in an ointment. The simultaneous determination was performed using isocratic reversed-phase mode within 20 min by employing an octyl (C8) column and a mobile phase containing sodium dodecyl sulfate (SDS) and 2-propanol. The method was successfully applied to quality control and stability testing of the ointment. PMID- 9226577 TI - Enantioseparation of beta-blockers labelled with a chiral fluorescent reagent, R (-)-DBD-PyNCS, by reversed-phase liquid chromatography. AB - A fluorescent chiral tagging reagent, 4-(3-isothiocyanatopyrrolidin-1-yl)-7-(N,N dimethylaminosulfony l)-2,1, 3-benzoxadiazole [R(-)-DBD-PyNCS], has been used for the liquid chromatographic resolution of racemic pairs of beta-blockers. The reagents reacts with beta-blockers at 65 degrees C for 90 min in aqueous acetonitrile containing 0.05% triethylamine to produce the corresponding pair of diastereomers. No racemization occurs during the tagging reaction under these conditions. From results of the time-course study of oxprenolol the reactivities of the enantiomers of beta-blockers with R(-)-DBD-PyNCS are comparable. The optimum excitation and emission wavelengths of the resulting derivatives were ca. 460 and 550 nm, respectively. The derivatives of beta-blockers were efficiently resolved by a reversed-phase column with water-acetonitrile containing 0.1% trifluoroacetic acid as the eluent. The resolution (Rs) values of the diastereomers derived from 10 beta-blockers were in the range of 1.54-4.80. The Rs value for timolol was 0.643. The detection limits (signal-to-noise ratio of 2) were one or two orders of magnitude lower with beta-blockers having the iso propylamino structure (15-300 fmol) than with those having the tert-butylamino structure (1.25-8.00 pmol). The proposed procedure was applied to the determination of R(+)- and S(-)-propranolol in rat plasma and saliva after oral administration of R(+)-propranolol hydrochloride or S(-)-propranolol hydrochloride. PMID- 9226578 TI - Direct observation of the separation process by the chromato-videoscope. AB - Separation process in a liquid chromatographic column were visualized and analyzed by a developed chromato-videoscope. The migration aspects were evaluated with successively obtained densitograms. In reversed-phase chromatography, the band widths of each solute band were almost equal fore both weakly and strongly retained solutes when compared at the same column position (the same migration distance in the column). In the gradient elution mode, the position of the solute band showed that the migration velocity of the solute band changed gradually according to changes in solvent composition. The drug trapping process to BSA- coated ODS packings for direct injection of biological fluid was also observed. In the absence of BSA from the sample solution, the drug molecules were trapped in a narrow band. However, at higher BSA concentrations in the sample solution, a broader band shape was observed. This band broadening shows how the drug molecules were retained on the protein. PMID- 9226579 TI - Video-microscopy for analysis of molecular dynamics in cells. AB - Real-time analysis of molecular dynamics in living cells was studied by developed video-microscopes. Two new detective methods were reported, one is for analysis of ciliary movement and the other is the qualitative analysis of exocytosis of insulin-containing granules with a video-enhanced light/fluorescent microscope. For analysis of ciliary movement, glass beads were migrated in the flow. The migration speed parallel to the flow produced by ciliary beating was used as an index of the beating activity. When tracheal epithelium isolated from mouse was incubated with ambroxol, and expectorant known to activate ciliary beat frequency, the floating speeds of glass beads were changed with 1 min of incubation. The results suggest that the present method is useful not only for screening of expectorants but also for the study of molecular mechanisms underlying ciliary beat of tracheal epithelium. Visualization of the moment of the release of contents from insulin-containing granules was achieved using video enhanced fluorescent microscopy in MIN6 cells of mouse insulinoma cell line. A fluorescent amino acridine dye, quinacrine, was found to be incorporated into low pH secretory granules, including insulin, in the cells. The granules which incorporated quinacrine emitted a slightly blue-green fluorescence. Upon stimulation with glucose, release of the quinacrine fluorescence from granules were observed. The present method would be useful for quantitative analysis of secretion of insulin from MIN6 cells as well as pancreatic beta-cells. PMID- 9226580 TI - Time-resolved fluoroimmunoassay for pituitary adenylate cyclase activating polypeptide 27 (PACAP27) using europium (III) ion chelate labeled streptavidin biotin complex. AB - Pituitary adenylate cyclase activating polypeptide (PACAP) is a novel peptide hormone and has a variety of biological action. In studies of the physiological behaviour of endogenous PACAP, the determination of PACAP levels in biological materials require a highly sensitive and specific method. Therefore, we developed a sensitive time-resolved fluoroimmunoassay (TR-FIA) for PACAP27 which is the biologically important fragment of PACAP. Accordingly, we developed TR-FIA using a biotinylated PACAP27 (b-PACAP27) as a tracer and europium (III) chelate labeled streptavidin-biotinylated bovine serum albumin complex as a detection of biotin on solid phase. A measurable range of PACAP27 was 7.8-1000 pg m1-1 by the proposed TR-FIA. For measurement of biological samples, the samples were purified to eliminate substances which interfered with the TR-FIA. The mean recovery of PACAP27 using commercially reversed phase column was 74.7% (n = 12). The various tissues, extracts and plasma concentrations of rat could be measured by the proposed TR-FIA. PMID- 9226581 TI - A novel high-performance liquid chromatographic assay for vitamin D metabolites using a coulometric electrochemical detector. AB - A new, highly sensitive HPLC assay method using an electrochemical detector (ECD) for multiple assay of vitamin D metabolites is reported. The assay involves extracting lipids from plasma with methylene chloride and methanol, purification on Zorbax SIL column with 5.5% (v/v) iso-propanol in hexane and quantification by HPLC-ECD. A coulometric system, composed of the dual electrode analytical cell and a guard cell, was used for ECD of the eluting compounds. The potentials applied to detectors 1 and 2 in a dual electrode analytical cell were adjusted to +0.20 V and +0.60 V, respectively. This method is sensitive to 20 pg of 25 hydroxyvitamin D3 [25(OH)D3] and of 24R,25-dihydroxyvitamin D3 [24,25(OH)2D3]. Calibration curves gave linearity from 20-1000 pg for 25(OH)D3 and 24,25(OH)2D3. The detection limit was approximately 50 pg ml-1 for 25(OH)D3 and 24,25(OH)2D3 in plasma. This sensitivity combined with an overall recovery of 25(OH)D3 (81.5 +/- 2.6%, mean +/- S.E.) allows the measurement of trace amount of 25(OH)D3 with only 20 microliters of plasma. Intra- and interassay RSD values were 5.3 and 9.7% for 25(OH)D3 and 6.3 and 9.7% for 24,25(OH)2D3, respectively. Plasma levels of 25(OH)D3 and 24,25(OH)2D3 in normal adults were 15.9 +/- 2.8 ng ml-1 (n = 10) and 1.4 +/- 0.5 ng ml-1 (n = 10), respectively. This method allows the determination of 25(OH)D2 and 25(OH)D3 for evaluating their nutritional and clinical status. From these results, it is concluded that the proposed HPLC-ECD assay system is useful for the determination of vitamin D metabolites in biological fluids as a highly sensitive physicochemical method. PMID- 9226582 TI - Micellar electrokinetic capillary chromatography of benzodiazepine antiepileptics and their desmethyl metabolites in blood. AB - A fast and reliable method for the MEKC separation and determination of benzodiazepine antipileptics and their active metabolites in serum has been developed, using a separation buffer composed of borate (pH 9.5), SDS (18 mM), and acetonitrile (14%) as an organic modifier. The method is sensitive enough to be used clinically with a precision of less than 3% for the analysis of nitrazepam, clonazepam, clobazam, diazepam and their desmethyl metabolites in serum. PMID- 9226583 TI - Determination of p-hydroxymandelic acid enantiomers in urine by high-performance liquid chromatography with electrochemical detection. AB - High-performance liquid chromatography (HPLC) with electrochemical detection using a chiral ligand-exchange column was developed for the enantioselective determination of p-hydroxymandelic acid (HMA), a metabolite of synephrine, with high sensitivity. A good linear relationship between current ratio and amount was noted for 0.5-500 pmol HMA, with a correlation coefficient of 0.999 for each HMA enantiomer. The relative standard deviation (R.S.D.) was 1.6% at 100 pmol d-HMA and 2.2% at 100 pmol l-HMA. The detection limit of each HMA enantiomer was 0.5 pmol (signal to noise ratio, S/N = 3). By this method, HMA in Citrus unshiu and in urine following the ingestion of C. unshiu was determined. Although no HMA was found in c. unshiu, d- and l-HMA were present in urine after the ingestion of C. unshiu. The time courses of HMA and conjugated synephrine enantiomers excreted in urine following the ingestion of C. unshiu for 24 h could be monitored. This method should prove applicable to the study of synephrine metabolism. PMID- 9226584 TI - Determination of acid values of fats and oils by flow injection analysis with electrochemical detection. AB - A new method using a flow injection system with electrochemical detection was developed to determine acid values of fats and oils. VK3 (2-methyl-1,4 naphthoquinone) solution, i.e., ethanol containing 3 mM VK3 and 38 mM LiClO4, was used as the carrier solution. Flow signals were monitored at -0.33 V vs. Ag/AgCl. For preparation of a sample solution, an oil sample was completely dissolved in VK3 solution, or fatty acids were extracted from the sample into this solution. Aliquots (5 microliters) of the sample solution were injected into the flow injection system. Acid values were determined based on flow signals for 14 samples and the results were found to be consistent with those by potentiometric titration. Relative standard deviation was less than 2%. Samples were processed at the rate of 60 h-1. The stability of fish and cod liver oils was followed by measuring acid values for 8 weeks. This method proved to be a simple and rapid means for acid value determination. PMID- 9226585 TI - Analysis of ibuprofen metabolites by semi-microcolumn liquid chromatography with ultraviolet absorption and pulsed amperometric detectors. AB - Semi-microcolumn (1.5 mm i.d.) liquid chromatography (LC) system with ultraviolet (UV) and pulsed amperometric detector (PAD) was constructed for analysis of ibuprofen metabolites in human urine. PAD was connected in series with the UV detector, and an alkaline solution was post-column added to a mobile phase after the UV detector. By a gradient elution, five ibuprofen metabolites were detected with UV detection from 1 microliter of human urine at 3.5 h after the administration, and information concerning their glucuronation was simultaneously obtained by PAD response. PMID- 9226586 TI - Simultaneous enantiomeric determination of a gastroprokinetic agent mosapride citrate and its metabolite in plasma using alpha 1-acid glycoprotein HPLC column. AB - Mosapride citrate, a novel benzamide-type gastroprokinetic agent, is clinically prescribed as a racemate and is metabolized to its des-4-fluorobenzyl structure (M-1). In order to analyze simultaneously the enantiomers of mosapride and M-1 in plasma, a simple and reproducible high-performance liquid chromatographic (HPLC) method has been developed. The enantiomeric separation and determination were successfully achieved using an alpha 1-acid glycoprotein column and gradient elution with a fluorimetric detection (excitation 314 nm/emission 352 nm). Both enantiomers of mosapride and M-1 were well separated between 20 and 22 min at pH 4.4 and between 4 and 7 min at pH 5.0, respectively. Accurate determinations are possible in the concentration ranges of 10-5000 ng ml-1 for mosapride enantiomers and 50-5000 ng ml-1 for M-1 enantiomers. The intra- and inter-day coefficients of variation are satisfactory for the pharmacokinetic study of mosapride. PMID- 9226587 TI - Analysis of single-strand conformation polymorphisms by capillary electrophoresis with laser induced fluorescence detection. AB - Detection of point mutations in genomic DNA is important for diagnosis of inherited characteristics and genetic diseases. A point mutation in a specific region of DNA amplified by polymerase chain reaction (PCR) can be detected with single-strand conformation polymorphism (SSCP) analysis. Analysis of SSCP by laser-induced fluorescence capillary electrophoresis in entangled polymer solution (CE-LIF) has been developed in the present paper. K-ras genes including seven mutations were amplified with primer labeled with Texas Red at its 5' end. The labeled PCR products were dissociated to single strands by heating and separated with capillary gel electrophoresis and He-Ne laser-excited fluorescence detection. Our results suggest that all fragments having normal (Gly) and mutated (Ala, Arg, Cys, Ser, Val, Asp) sequences at codon 12 can be distinguished. Analysis of SSCPs with CE-LIF is well suited for clinical analysis of SSCPs because of its high sensitivity, resolution, reproducibility and speed. PMID- 9226588 TI - Analysis of peptides and proteins by temperature-responsive chromatographic system using N-isopropylacrylamide polymer-modified columns. AB - A new method of HPLC using packing materials modified with a temperature responsive polymer, poly(N-isopropylacrylamide) (PIPAAm), was developed. Homogeneous PIPAAm polymer and its copolymer with butyl methacrylate (BMA) were synthesized and grafted to aminopropyl silica by activated ester-amine coupling and they were used as packing materials. The surface properties and functions of the stationary phases are controlled by external temperature. Isocratic elution by aqueous mobile phase alone is the basis for separation of peptides and protein. The separation of the mixture of three peptides, insulin chain A and B and beta-endorphin fragment 1-27 was achieved by changing the column temperature with 0.9% NaCl aqueous solution as the sole eluent. Retention of peptides and proteins was controlled both by column temperature and by NaCl concentration in the aqueous mobile phases in this chromatographic system. PMID- 9226589 TI - Survey and assessment of the actual state of routine measurement of glycohaemoglobin/GHb by commercial methods: warning to the users and the providers. AB - As the clinical availability of glycohaemoglobin/GHb measurement increases, so does the need for comparable and accurate values among different laboratories and different methods. At least there should be comparability, i.e., commutability or feasibility of providing comparable results from different assays in different laboratories. A clinical joint study on insulin therapy, a survey of the actual inter-laboratory differences in GHb measurement among 41 institutions and an assessment of 11 assay methods for the determination of GHb were performed using commercial calibrators and fresh blood samples. Data on the actual state of inter laboratory and inter-assay differences of observed values were useful for comparing results among facilities. The recommendation of the Japan Diabetes Society to measure only the stable GHb component and to correct the GHb percentage by two-point calibration with assigned values, was effective but not sufficient. Even after correction, 8 out of 11 methods still remained of little practical use because of their large relative errors. Inter-method differences among 11 available assay methods were great even after correction and depended on not only the methods but the samples used for the determination. The performance of some methods or instruments used are only poor at distinguishing the stable glycated haemoglobin itself. Some alternative measurement system with comparability, commutability and precision should be established. An urgent and worldwide problem to remove inter-laboratory differences in the measurement of GHb needs to be solved. Users in clinical practice must recognize these problems, and, before supply, the providers should check their method and keep records that are readily traceable. PMID- 9226590 TI - Instrumentation of a handy microscopic probe for concurrent observation and measurement of active sweat secretion, and its applications. AB - Instrumentation for the concurrent, dynamic monitoring of active sweat glands and perspiration volume is described. A device for the measurement of the rate of sweat secretion was installed on the head part of a microscope. The combined apparatus (microscopic probe) is handy for use and its weight is very light (ca. 300 g). The microscopic probe is easily attached to the surface of human skin. The dynamic activities of the sweat glands on the forehead and nose and under the nose were observed and measurement when thermal, mental and physical stimuli were applied. The activities of individuals sweat glands were asynchronous when observed in units of a few seconds or less; however, they worked synchronously in a unit period of several seconds. The latter were recorded as fine peaks by a strip chart recorder. The proposed system may be useful for the study of the sympathetic nervous system, the skin sympathetic reflex and the working of sudomotor nerves. PMID- 9226591 TI - The effect of single does of rifampicin on the pharmacokinetics of oral nifedipine. AB - The pharmacokinetic interaction of oral rifampicin (1200 mg) and oral nifedipine (10 mg capsules), given as single doses, was investigated in six healthy volunteers (mean age 28.5 +/- 6.3 years and mean weight 67.0 +/- 4/45 kg). The plasma concentrations of nifedipine was monitored using a HPLC technique, 8 h after pre-treatment, with rifampicin. The mean relative bioavailability of nifedipine following pre-treatment with rifampicin 1200 mg was 35.8% (P < 0.0001). The mean elimination half life (t1/2) of nifedipine decreased from 2.62 to 1.03 h (P < 0.0001); and, the total clearance (ClT) increased from 17.33 to 50.17 ml min-1 kg-1 (P < 0.0001). There were no significant differences in Vd and Tmax. The study suggests that the effect of induction by rifampicin decreases the bioavailability of nifedipine by either increasing the first pass effect or decreasing its oral absorption. The induction also increases the clearance of nifedipine. PMID- 9226592 TI - Selected applications of capillaries with dynamic or permanent anodal electroosmotic flow in chiral separations by capillary electrophoresis. AB - The techniques for a dynamic and permanent reversal of the electroosmotic flow (EOF) were used for the reversal of the enantiomer migration order (EMO) of neutral and cationic analytes in chiral capillary electrophoresis (CE). Native beta-Cd and an anionic CD derivative, CM-beta-CD were used in both, bare silica- and positively coated capillaries. Advantages and disadvantages of a dynamic and permanent modification of the capillary inner surface are briefly discussed. PMID- 9226593 TI - Experimental design strategies in the optimization and robustness testing of adsorptive stripping voltammetric conditions for kynurenic acid determination. AB - Experimental design was used for the optimization and robustness testing of an adsorptive stripping voltammetric procedure for kynurenic acid determination. The optimization of the peak height response proceeded through a screening phase (D optimal design strategy) followed by a response surface study (Doehlert design) applied to the variables pH, pulse amplitude and stirring rate. An interaction between pH and stirring rate was pointed out. The optimized method was validated and the variation of factors that was expected to occur in practice was simulated in a robustness test. A composite fractional matrix for the evaluation of method robustness was used and pH emerged as the only critical factor. The linear range found applying the optimized conditions was 2.5 x 10(-9) to 2.5 x 10(-7) M and the calculated limit of detection was 1.72 x 10(-9) M. PMID- 9226594 TI - Enantioselective protein binding of semotiadil and levosemotiadil determined by high-performance frontal analysis. AB - An on-line frontal analysis HPLC system was developed for the determination of the unbound concentrations of semotiadil, a new calcium antagonist with non dihydropyridine structure, and its antipode (Levosemotiadil), and was applied to the enantioselective investigation of their plasma protein binding properties. This system consists of a high-performance frontal analysis (HPFA) column, an extraction column, and an analytical column, which are connected via two switching valves. After the direct injection of the sample solution into the HPFA column, the drug was eluted as a zonal peak with a plateau region. The unbound drug concentration was determined as the drug concentration in the plateau. As low as 1.04 nM of the unbound drug was determined with good reproducibility. Semotiadil (R-isomer) and levosemotiadil (S-isomer) are bound strongly and enantioselectively to human serum albumin (HSA) and human alpha 1-acid glycoprotein (AGP), and the enantioselectivity was reversed between these plasma proteins. While HSA binds S-isomer more strongly than the antipode, human AGP binds R-isomer more strongly. In human plasma, the unbound drug fraction was less than 1%, and the enantioselectivity was similar to that observed in AGP solution. PMID- 9226595 TI - Use of 1H-NMR spectroscopy to determine the enantioselective mechanism of neutral and anionic cyclodextrins in capillary electrophoresis. AB - One-dimensional (ID) and two-dimensional (2D) 1H nuclear magnetic resonance (NMR) techniques have been used to investigate the chiral recognition process in capillary electrophoresis (CE) for seven different cyclodextrins (CDs) with the calcium channel blocker amlodipine as a model compound. These include five neutral CDs (alpha-CD, beta-CD, gamma-CD, hydroxypropyl-beta-CD and hydroxyethyl beta-CD) and two anionic CDs (sulphobutyl-ether-beta-CD and carboxymethyl-beta CD) where mixtures of amlodipine with each of the seven CDs were examined by 1D NMR in deuterated phosphate buffer at pD 3.4. The resonance shift of signals with added CD, relative to the CD-free position (shift displacement, delta delta) and shift non-equivalence (delta delta *) of enantiomeric signals shifted relative to each other after addition of CD were examined for non-overlapped protons of amlodipine. The possible correlations of NMR shift non-equivalence data with chiral separation in CE for amlodipine have been critically assessed. Qualitative differences in the 1D NMR shifts and enhanced enantioselectivity in CE were observed for amlodipine with sulphobutyl-ether-beta-CD. Further experiments on the through-space interactions using 2D rotating frame nuclear Overhauser effect spectroscopy (ROESY) indicated that there was no association between internal glucopyranose hydrogen atoms and the aromatic hydrogens of amlodipine. This gives evidence for the aromatic ring not being included in this CD. Moreover, data from spin-lattice relaxation times (T1) measured for amlodipine in the free state and after addition of the anionic sulphobutyl-ether-beta-CD indicate that the aromatic moiety of amlodipine is not included into the sulphobutyl-ether-beta-CD cavity. There is evidence that it interacts with the sulphobutyl side chains, and may adopt a preferred orientation outside the sulphobutyl-ether-beta-CD toroid itself. PMID- 9226596 TI - Simultaneous determination of creatinine, hypoxanthine and uric acid in biological samples by column-switching liquid chromatography with ultraviolet detection. PMID- 9226597 TI - Sleep pathophysiology--a useful tool in psychiatry. PMID- 9226598 TI - Sleep-related erections. PMID- 9226599 TI - Cardiovascular disease and sleep-related erections. AB - Sexual difficulties are highly prevalent in male patients with cardiovascular diseases, such as hypertension, atherosclerosis, and hypercholesterolemia. Recently, several studies have been conducted on the effects of cardiovascular diseases, as well as associated drug and nondrug treatments, on nocturnal penile tumescence (NPT) and other measures of sexual function. Although an overall trend has been observed toward decreased NPT in patients with chronic hypertension and other cardiovascular conditions, design and methodological difficulties have been noted in most studies, and results have been generally, inconclusive. Similarly, antihypertensive drugs such as beta-blockers and diuretics have been associated with diminished NPT in several studies, although methodological problems have again been noted. Furthermore, the mechanism of action of antihypertensive drugs on sleep-related erections has not been determined. Most recently, a positive effect of cholesterol-lowering drugs (pravastatin, lovastatin) on NPT has been observed in middle-aged males with chronic hypercholesterolemia. Additional studies of the effects of cardiovascular disease on NPT and other measures of sexual function are needed. PMID- 9226600 TI - Evaluation of erectile dysfunction and sleep-related erections. AB - Significant advances in this past decade have improved our understanding of erectile physiology. A variety of tests are available for diagnosing impotence. SRE testing provides objective physiological information that is useful for indexing erectile capability and formulating a rational treatment plan. As such, SRE testing is a powerful noninvasive tool for assessing dysfunction. Nonetheless, in making a final diagnosis, the skillful clinician relies on more than one assessment parameter and on clinical acumen. PMID- 9226601 TI - Androgen and sleep-related erections. AB - Sleep-related penile erections provide a unique opportunity to objectively study erectile physiology in man. Testosterone is one of several factors involved in normal sexual function and testosterone reduction can be achieved by administering luteinizing-hormone releasing-hormone agonists (LHRH-A). In this study, ten healthy, young adult males were administered LHRH-A or placebo for a 12-week period. Subjects taking LHRH-A had a marginally significant decline in sleep-related erection duration at week 4 and significant reductions at weeks 8 and 12. By contrast, no statistically reliable change was found for the number of erections over the course of study. Maximum circumference increase during sleep erections showed mixed results. These results indicate that, whereas androgen reduction adversely affects sleep-related erections, it does not eliminate them over a 12-week trial in healthy young adult men. Further study in a larger sample is needed. Nonetheless, these preliminary findings support androgen having an important role in sleep-related erections. PMID- 9226602 TI - Sleep-related erections: absence of change following presleep sexual arousal. AB - We examined the effects of a brief period of sexual arousal before sleep on sleep related erections (SREs) to add to our knowledge concerning those factors that affect SREs. Twelve subjects watched a 5 minute sexually explicit video before sleep. On other evenings they watched a dysphoric arousal video or a lecture (neutral) video. Sleep and SREs were recorded throughout the following night. Although the brief sexual arousal video produced a full or near full erection in all subjects, no significant effect on subsequent SREs occurred. We conclude that the control of SREs in young healthy subjects is insulated against the effect of a brief period of sexual arousal before sleep. PMID- 9226603 TI - The link between sleep and depression: the effects of antidepressants on EEG sleep. AB - The assumption that sleep dysregulation is more than a mere epiphenomenon of depression is based on several observations: sleep disturbances are strongly associated with the depressive state; a number of sleep manipulations can alleviate symptoms of depression in some patients; and the majority of antidepressants bring about remarkable changes in sleep polygraphic variables. An obvious question is whether changes in sleep physiological processes are intimately involved in the pathogenesis and recovery from depression. One way to elucidate the link between sleep and depression is to examine whether the influence of antidepressants on sleep is related to clinical improvements in depressives. For that purpose, the effects of antidepressants on EEG sleep and their importance for the treatment of depression are summarized against the background of two existing hypotheses concerning the link between sleep and depression: one hypothesis concerning the role of REM; the other concerning the role of non-REM sleep. EEG sleep studies on the use of antidepressants in depressives have not produced clear evidence of the involvement of REM sleep or non-REM sleep in the mechanisms underlying clinical change. Furthermore, the role of sleep physiological mechanisms during treatment with antidepressants is still unclear. To interpret the effects of antidepressants on EEG sleep in terms of sleep physiological processes more fundamental sleep research is necessary. Also, more comparative studies of antidepressants with similar therapeutic effects but different pharmacological profiles are needed in both healthy and depressed subjects to further quantify the impact of EEG sleep modification in the recovery from depression and to differentiate between pharmacological and sleep-related aspects. PMID- 9226604 TI - REM sleep in depressed patients: different attempts to achieve adaptation. AB - Twenty-seven depressed patients and 10 healthy subjects were investigated in the sleep laboratory during two to three consecutive nights. Eleven of the 27 patients demonstrated the "first night effect" (group I) and 11 other patients demonstrated a clear absence of the "first night effect" (group II). Five of the 27 depressed patients were omitted from the study because they did not fit criteria for first night effect. The 10 healthy controls demonstrated a first night effect. In group I, the duration of the first rapid eye movement (REM) sleep episode was increased on the first night and on the second night the REM sleep latency was decreased, whereas REM sleep duration and eye movement (EM) density was increased. The number of the short sleep cycles (less than 40 minutes) was greater in group I versus group II and the percentage of slow-wave sleep (SWS) was also higher in group I. In depressed patients with the "first night effect" the enhanced REM sleep requirement is satisfied not only by an increased REM sleep duration but also by the improved REM sleep quality that is crucial for adaptation. The adaptive role of the increased first REM period and the increased EM density in this period is very limited. PMID- 9226605 TI - Actigraphic sleep monitoring in posttraumatic stress disorder (PTSD) patients. AB - Posttraumatic stress disorder (PTSD) patients frequently complain that they suffer from sleep disturbances. To date, the polysomnographic studies that have attempted to study PTSD patients' subjective complaints of sleep difficulties have produced conflicting results. The objective of the present study was to compare PTSD patients' subjective complaints of poor sleep and objective actigraphic recordings of their sleep over a period of several consecutive nights. The results indicate that PTSD patients do not suffer from poorer sleep than a control group, based on actigraphic measures, and that their subjective sleep evaluation is inconsistent with objective sleep measures. These patients fail to correctly estimate their sleep. PMID- 9226606 TI - Sleep quality versus sleep quantity: relationships between sleep and measures of health, well-being and sleepiness in college students. AB - Two studies assessed whether measures of health, well-being, and sleepiness are better related to sleep quality or sleep quantity. In both studies, subjects completed a 7-day sleep log followed by a battery of surveys pertaining to health, well-being, and sleepiness. In subjects sleeping an average of 7 hours a night, average sleep quality was better related to health, affect balance, satisfaction with life, and feelings of tension, depression, anger, fatigue, and confusion than average sleep quantity. In addition, average sleep quality was better related to sleepiness than sleep quantity. These results indicate that health care professionals should focus on sleep quality in addition to sleep quantity in their efforts to understand the role of sleep in daily life. PMID- 9226608 TI - Posttraumatic stress disorder, tenderness and fibromyalgia. AB - The aims of the present study were to inquire into the prevalence of fibromyalgia syndrome, to assess nonarticular tenderness, to measure fibromyalgia syndrome related symptoms, quality of life, and functional impairment among posttraumatic stress disorder (PTSD) patients as compared with control subjects. Furthermore, the differences between the PTSD patients with and without fibromyalgia syndrome were studied. Twenty-nine PTSD patients and 37 control subjects were assessed as to the diagnosis of fibromyalgia syndrome according to the American College of Rheumatology. Tenderness was assessed manually and with a dolorimeter. Fibromyalgia syndrome-related symptoms, quality of life, physical functioning, PTSD symptomatology, and psychiatric features were assessed by valid and reliable self-report inventories. Results showed that the prevalence of fibromyalgia syndrome in the PTSD group was 21% vs. 0% in the control group. Furthermore, the PTSD group was more tender than the control group. PTSD subjects suffering from fibromyalgia syndrome were more tender, reported more pain, lower quality of life, higher functional impairment and suffered more psychological distress than the PTSD patients not having fibromyalgia syndrome. It is suggested that previous reports on diffuse pain in PTSD in fact described undiagnosed fibromyalgia syndrome. The link between psychological stress and pain syndromes is emphasized. PMID- 9226607 TI - The relation of sleep difficulties to fatigue, mood and disability in chronic fatigue syndrome. AB - The relationship of sleep complaints to mood, fatigue, disability, and lifestyle was examined in 69 chronic fatigue syndrome (CFS) patients without psychiatric disorder, 58 CFS patients with psychiatric disorder, 38 psychiatric out-patients with chronic depressive disorders, and 45 healthy controls. The groups were matched for age and gender. There were few differences between the prevalence or nature of sleep complaints of CFS patients with or without current DSM-IIIR depression, anxiety or somatization disorder. CFS patients reported significantly more naps and waking by pain, a similar prevalence of difficulties in maintaining sleep, and significantly less difficulty getting off to sleep compared to depressed patients. Sleep continuity complaints preceded fatigue in only 20% of CFS patients, but there was a strong association between relapse and sleep disturbance. Certain types of sleep disorder were associated with increased disability or fatigue in CFS patients. Disrupted sleep appears to complicate the course of CFS. For the most part, sleep complaints are either attributable to the lifestyle of CFS patients or seem inherent to the underlying condition of CFS. They are generally unrelated to depression or anxiety in CFS. PMID- 9226609 TI - Chronic fatigue syndrome: a qualitative investigation of patients' beliefs about the illness. AB - The chronic fatigue syndrome is a disabling chronic condition of uncertain cause. Previous studies have found that patients seen in hospital clinics with the syndrome often strongly believe that their illness is physical in nature and minimize the role of psychological and social factors. There is also evidence that patients cope by avoiding activity. However, almost all of these studies have assessed illness beliefs only by questionnaire. The aim of this study was to explore the nature and origin of illness beliefs in more detail using in-depth interviews and a qualitative analysis of patient responses. Sixty-six consecutive referrals meeting Oxford criteria for chronic fatigue syndrome were recruited. Analysis of responses indicated that, whereas the most commonly described explanation for the illness was a physical one, more than half the patients also believed "stress" had played a role. Patients believed that they could partially control the symptoms by reducing activity but felt helpless to influence the physical disease process and hence the course of the illness. Patients reported that they had arrived at these beliefs about the illness after prolonged reflection on their own experience combined with the reading of media reports, self help books, and patient group literature. The views of health professionals played a relatively small role. There is potentially a considerable opportunity to help patients arrive at a wider and more enabling explanation of their illness when they first present to primary care. PMID- 9226611 TI - Basic principles and techniques of pharmacokinetic modeling. AB - Pharmacokinetics has been extensively utilized to calculate safe and effective drug dosages for many exotic animals species. This practice is optimal when the complete pharmacokinetic profile is determined from an intravenous study in the target species. Extrapolation across species using allometric techniques also require precise yet robust intravenous pharmacokinetic data from the species for which the analyses is made. The report is an overview of the design and analysis of such pivotal pharmacokinetic experiments, taking into consideration the use of pilot studies to determine optimal sampling times, the true purpose of the study (determine model structure vs. estimating a known model's parameters), and strategies of data analysis. By following these simple guidelines, the exotic animal community should benefit by having reliable data on drug disposition for their species of interest. PMID- 9226612 TI - Toxicant use in the zoo environment. AB - Toxicants, such as insecticides, rodenticides, caging material, and cleaning agents, are used on a daily basis in zoos to provide a healthy environment. These products must be used carefully so that the zoo inhabitants are not inadvertently poisoned. Product labels should be read, directions followed, and warnings heeded. An understanding of the mechanisms of action of the products as well as their effects on different species will assist zoo personnel in diagnosis and treatment decisions should toxicosis occur. Information regarding the effects of toxicants in exotic species is limited and so much information must be extrapolated from information on domestic species. PMID- 9226613 TI - Lead and zinc intoxication in zoological medicine: a review. PMID- 9226614 TI - Pharmacokinetics of a single intravenous enrofloxacin dose in scimitar-horned oryx (Oryx dammah). AB - Based on a 1.3 mg/kg mean dosage determined by metabolic energy scaling, enrofloxacin pharmacokinetics of a single i.v. dose of enrofloxacin in five adult scimitar-horned oryx (Oryx dammah) were determined. Drug concentration versus time curves were best fit by residual analysis to a one-compartment open model with a maximum (mean +/- SD) serum concentration after distribution of 1.887 +/- 0.632 micrograms/ml and an elimination half-life of 41.2 +/- 27.5 min. Model independent parameters were area under the curve (173.63 +/- 147.5 micrograms.min/ml), mean volume of distribution (steady state) (0.80 +/- 0.30 L/kg), clearance (12.07 +/- 7.12 ml/min/kg), and residence time (77.22 +/- 72.8 min). Mean serum enrofloxacin concentrations reached the recommended minimum inhibitory concentration (1.0 micrograms/ml). Drug concentrations remained above the minimum inhibitory concentration of most sensitive bacteria (0.5 micrograms/ml) consistently for 90 min. Based on this study, enrofloxacin would have to be administered parenterally to scimitar-horned oryx at 1.6 mg/kg every 6 8 hr (minimally) to maintain appropriate serum concentrations against susceptible bacteria. The metabolic energy scaled dosed regiment from this study appeared to be too low for the oryx. PMID- 9226615 TI - Disposition of single-dose intravenously administered enrofloxacin in emus (Dromaius novaehollandiae). AB - The pharmacokinetics of enrofloxacin in emus (Dromaius novaehollandiae) were examined following parenteral administration. A mean allometrically scaled dose of 2.2 +/- 0.03 mg/kg was administered as a single i.v. bolus, and serum samples were collected at predetermined intervals over a 24-hr period. Enrofloxacin levels were measured using high-performance liquid chromatography, and the resulting concentration versus time curve was analyzed using nonlinear regression with least squares parameter estimation. The data were best represented by a two compartment model with a mean elimination half-life of 3.33 hr. Mean model independent parameters obtained were area under the curve (8.26 micrograms.hr/ml), mean residence time (4.40 hr), apparent volume of distribution (1.49 L/kg), and total body drug clearance (0.36 L/hr/kg). Mean serum concentrations exceeded the target peak of 2.0 micrograms/ml and remained above an estimated inhibitory concentration of 0.5 micrograms/ml for approximately 2 hr. Based upon the results of this study, enrofloxacin administered parenterally to emus at 2.2 mg/kg every 12 hr is expected to achieve therapeutic serum concentrations against susceptible organisms. PMID- 9226616 TI - Disposition of single-dose intravenously administered amikacin in emus (Dromaius novaehollandiae). AB - The pharmacokinetics of amikacin in emus (Dromaius novaehollandiae) was examined following parenteral administration. A mean 7.2 +/- 0.12 mg/kg dose was administered as a single i.v. bolus, and serum samples were collected at predetermined intervals over a 24-hr period. Amikacin levels were measured using a fluorescence polarization immunoassay, and the resulting concentration-versus time curve was analyzed using nonlinear regression with least squares parameter estimation. The data were best represented by a three-compartment model with a mean elimination half-life (t1/2 beta) of 0.87 hr, with a longer rate of elimination from the third compartment (t1/2 gamma = 6.06 hr). Mean model independent parameters obtained were area under the curve (269.66 micrograms.hr/ml), mean residence time (6.48 hr), apparent volume of distribution (0.18 L/kg), and total body drug clearance (0.03 L/hr/kg). Mean serum concentrations exceeded a target peak of 32.0 micrograms/ml and remained above an estimated inhibitory concentration of 8.0 micrograms/ml for approximately 12 hr. Mean serum levels had declined below a target trough of 4 micrograms/ml at 24 hr. PMID- 9226617 TI - The pharmacokinetics of a single intramuscular dose of amikacin in red-tailed hawks (Buteo jamaicensis). AB - The pharmacokinetic parameters of amikacin were determined in red-tailed hawks (Buteo jamaicensis) following the i.m. administration of a single 20 mg/kg dose. After a rapid absorption phase, mean amikacin serum concentrations peaked at 65 +/- 12 micrograms/ML 30-45 min following injection. The serum amikacin concentrations decreased to 2.3 +/- 2 micrograms/ml at 12 hr postinjection. Amikacin was eliminated with first-order kinetics characteristic of a single compartment model with a half-life of 2.02 +/- 0.63 hr. The volume of distribution was estimated to be 0.28 +/- 0.03 L/kg. Forty-two isolates of gram negative bacteria and coagulase-positive Staphylococcus species were cultured from birds of prey presented to the Veterinary Medical Teaching Hospital at the University of California-Davis. The minimum inhibitory concentration (MICs) of amikacin ranged from 0.5 to 8.0 micrograms/ml (mean = 2.5 micrograms/ml). The 20 mg/kg dose used in this study resulted in serum concentrations at or above the MICs for > 12 hr for most of the isolates examined. The heaviest birds had the lowest peak serum amikacin concentrations, and the lightest birds had the highest, despite exact volume replacement for each sample drawn. This observation suggests that doses should be based on factors other than weight alone. Amikacin administered at 15-20 mg/kg/day, either as a single dose or divided into two or three doses, is effective in treating sensitive pathogens of the red-tailed hawk. PMID- 9226618 TI - Mercury distribution in American alligators (Alligator mississippiensis) in Florida. AB - Thirty American alligators (Alligator mississippiensis), including 24 wild-caught and six control captive farm-raised alligators, were analyzed for whole body mercury contamination. Wild-caught animals were collected from Water Conservation Area 3 in the Everglades ecosystem (n = 12) and from Alachua, Brevard, and Collier counties outside the Everglades (n = 12). Using cold-vapor atomic absorption spectrophotometry, samples of brain, cervical spinal cord, liver, paired kidneys, paired testes, paired ovaries, paired oviducts, heart, lungs, spleen, bile, tail and leg muscle, and tail and leg scales were analyzed on a wet weight basis to determine mercury concentration. Mercury was consistently detected in all specimens except for bile. Farm-raised alligators, fed a commercially prepared diet, contained very low mercury concentrations in all tissues analyzed. In comparison with alligators from outside the Everglades, Everglades alligators had significantly elevated concentrations of mercury in all tissues analyzed except ovaries, oviduct, bile, tail scales, and leg scales (paired two-sample Student's tau-test, P < 0.05). Muscle concentrations exceeded state (0.50-1.50 ppm) and federal (1.00 ppm) allowances for safe human consumption in alligators collected in the Everglades. No clinical signs of neurologic, hepatic, or renal toxicosis were detected. Because of the alligator's ability to bioaccumulate mercury, this species might be useful as a bio-monitor for environmental mercury contamination. PMID- 9226619 TI - Disposition of enrofloxacin and its metabolite ciprofloxacin after intramuscular injection in juvenile Burmese pythons (Python molurus bivittatus). AB - Eleven juvenile Burmese pythons (Python molurus bivittatus) weighing 0.75-1.75 kg were randomly divided into two groups. Blood samples were obtained through surgically placed anterior carotid artery cannulas. Six pythons received a single i.m. injection of enrofloxacin at 5 mg/kg. Blood samples were obtained at 0.5, 1, 3, 6, 12, 24, 48, 72, and 96 hr postinjection. A mean (+/- SD) maximal plasma concentration of 1.66 (+/- 0.42) micrograms/ml was measured at 5.75 hr postinjection. The harmonic mean half-life was calculated to be 6.37 hr. The second group of five snakes received enrofloxacin at 5 mg/kg i.m. s.i.d. for 5 days. Blood was collected immediately before each injection and at 6 hr after each injection. Over the 5-day period, there was a stepwise increase in mean trough plasma concentrations of enrofloxacin. Clinically effective peak plasma enrofloxacin concentrations were attained after the first injection but did not significantly increase during the sampling period. Pharmacokinetic data were assessed against minimum inhibitory concentrations of enrofloxacin for Pseudomonas ssp. isolates in snakes obtained from historical data at the Veterinary Medical Teaching Hospital, University of Florida. Enrofloxacin should be administered at 10 mg/kg i.m. every 48 hr when treating Pseudomonas ssp. infections in juvenile Burmese pythons. Treatment of infections of more enrofloxacin-sensitive gram-negative bacteria could be achieved with the administration of an initial i.m. dose of 10 mg/kg followed by 5 mg/kg every 48 hr. PMID- 9226620 TI - Amikacin pharmacokinetics and the effects of ambient temperature on the dosage regimen in ball pythons (Python regius). AB - The serum concentration of amikacin following intracardiac and i.m. administration of amikacin (3.48 mg/kg) in 12 ball pythons (Python regius) housed at 25 degrees C and 37 degrees C was studied. Blood samples were collected by cardiocentesis at intervals up to 144 hr after administration of amikacin. Drug concentration-versus-time curves following intracardiac administration at both temperatures best fit a two-compartment open model. For snakes housed at 37 degrees C, the extrapolated time 0 concentration (mean +/- SD) was 17.64 +/- 3.5 micrograms/ml with a median elimination half-life of 4.5 days. The maximum concentrations were 11.98 +/- 1.67 micrograms/ml and 13.87 +/- 2.61 micrograms/ml for snakes housed at 25 degrees C and 37 degrees C respectively. There were no significant pharmacokinetic differences among the snakes housed at 25 degrees C and 37 degrees C. Model-independent parameters were area under the curve, 69,900 +/- 0.011 micrograms.min/ml, apparent volume of distribution at steady state, 410 +/- 106 ml/kg, clearance, 0.036 +/- 0.009 ml/min/kg, and mean residence time, 3,530 +/- 273.7 minutes. Mean serum amikacin concentrations did not reach the recommended therapeutic peak concentrations for mammals (25 micrograms/ml). In addition, the amikacin serum concentration did not fall below the recommended therapeutic trough concentrations (2 micrograms/ml) by 6 days. The serum amikacin concentrations were efficacious based on the area under the curve. Therefore, amikacin (3.48 mg/kg) administered i.m. to ball pythons should produce maximum serum concentrations against most pathogenic bacteria. In this study, it would have taken another half-life, or 4.5 days, before trough concentrations of 2 micrograms/ml were achieved. To prevent accumulation, a one-time administration of amikacin may be appropriate. PMID- 9226622 TI - Safety of milbemycin as an oral or bath treatment for the tropical freshwater angelfish Pterophyllum scalare. AB - Technical grade milbemycin (A3-A4 oxime) was formulated in propylene glycol to produce a stock concentration of 5.0 mg/ml. Groups of six pond-reared freshwater angelfish (Pterophyllum scalare) randomly housed in 32-L aquaria were exposed to milbemycin by prolonged bath at 63, 125, and 188 PPB or by its incorporation into their gelatinized food at 2.5 mg or 5.0 mg/100 g food, which they were fed ad lib. for 1 day. Control fish were exposed to a prolonged bath (24 hr without charcoal filtration) of 0.8 ml propylene glycol/32 L water, were given gelatinized food incorporating 1 ml propylene glycol/100 g food ad lib. for 1 day, or were untreated (no propylene glycol exposure). All fish treated at 188 PPB and the smallest individuals from the 63- and 125-PPB aquaria died. Other fish at 125 PPB exhibited transient lethargy and increased opercular movement but recovered within 24 hr. No deleterious effects were noted in the fish given milbemycin orally. Pretreatment parasitic nematode infection rate, evaluated by gut dissection of 16 randomly selected fish, was 68.75%. Identification of nematodes to species was not performed. No significant differences in infection rates between treated and untreated groups were detected. PMID- 9226621 TI - Itraconazole plasma and tissue concentrations in the spiny lizard (Sceloporus sp.) following once-daily dosing. AB - Mycotic infections in reptiles present as primary diseases and as secondary problems in healing wounds and immunocompromised animals. A triazole antimycotic drug, itraconazole is orally active and well distributed and is effective against many common fungal pathogens in humans. To assess plasma and tissues concentrations after oral dosing in reptiles, a 23.5-mg/kg (mean) itraconazole dose was administered orally with a standard food bolus once daily for 3 days to 10 groups of three or four spiny lizards (Sceloporus sp.). On days 0, 1, 2, 3, 4, 6, 9, 12, and 18, group samples of blood, liver, and muscle were collected. Microbiologic assay of itraconazole concentrations was performed on these pooled samples. Values from an elimination graph of the concentrations of area under the curve (377.21 micrograms.hr/ml) and terminal elimination half-life (48.3 hr) were obtained for itraconazole in spiny lizard plasma. Peak itraconazole concentration of 2.48 micrograms/ml was obtained in two half-lives and would be expected to achieve steady state at approximately 3.1 micrograms/ml plasma concentration in 10 days. Peak liver concentration of 4.27 micrograms/ml was attained in 89.95 hr. Muscle concentration did not exceed 0.63 micrograms/ml and declined by 97.3 hr. With this dosing regimen, itraconazole plasma and liver concentrations would persist within reported minimum inhibitory concentrations for many fungal pathogens for 6 days beyond the peak concentration. PMID- 9226623 TI - Organochlorine pesticides associated with ocular, nasal, or otic infection in the eastern box turtle (Terrapene carolina carolina). AB - From May 1987 to September 1994, 19 eastern box turtles (Terrapene carolina carolina) originating from scattered locations on Long Island, New York (USA) were presented with one or more of the following signs: listlessness, ocular and nasal discharge, conjunctivitis, blepharitis, and otitis media. Numerous species of bacteria and yeast were isolated by aerobic culture. Histopathologic findings confirmed chronic active bacterial infections. Toxicologic analyses of livers revealed elevated concentrations of chlordane metabolites in two diseased turtles. A third turtle liver contained residues of endosulfan sulfate. Immunosuppressive effects of low-level exposure to organochlorines, including chlordane and endosulfan, could be involved in the pathogenesis of the observed infections. PMID- 9226624 TI - Zinc toxicosis in a Celebes ape (Macaca nigra) following ingestion of pennies. AB - An adult Celebes ape (Macaca nigra) was presented for a routine yearly evaluation. Abdominal radiographs revealed four radiodense metallic foreign objects resembling coins within the gastrointestinal tract. Hematologic abnormalities included a mild nonregenerative anemia and a mild leukocytosis, and the biochemical profile reflected renal dysfunction. Serum zinc levels were elevated. The animal was denied food for 24 hr in preparation for the endoscopic removal of the foreign objects. During this period, two partially eroded pennies were passed in the feces. Endoscopy was temporarily postponed, and the animal was offered food. Two days later a third penny was passed. When the fourth coin did not pass within the next 2 days, the animal was once again denied food, and the fourth coin was passed within 24 hr. Because of the temporal association between the fasting cycles and the passage of the coins, a causal relationship is suggested. Following the passage of all coins, serum zinc levels decreased, and hematologic and serum biochemical abnormalities improved. Six months later, all abnormalities had resolved. PMID- 9226625 TI - Presumptive red maple (Acer rubrum) toxicosis in Grevy's zebra (Equus grevyi). AB - Two female Grevy's zebras (Equus grevyi), one juvenile and one adult, were treated for hemolytic anemia. The juvenile survived, but the adult animal, which also had methemoglobinemia, was euthanized after it failed to recover from anesthesia. Significant pathologic findings in the adult zebra included generalized icterus, hemoglobinuric nephrosis, and paracentral hepatic necrosis. Serum titers for known infectious causes of anemia were negative. Examination of the zebra holding areas revealed two hybrid red maple (Acer sp.) trees. There was no known exposure to other hemolytic agents. This is the first report of probable red maple-induced hemolysis in zebra. PMID- 9226626 TI - Lead poisoning and intestinal perforations in a snapping turtle (Chelydra serpentina) due to fishing gear ingestion. AB - An adult male snapping turtle (Chelydra serpentina) was presented to the Tufts Wildlife Clinic with generalized weakness and limited ability to walk. A fishing hook was lodged in the corner of its mouth, monofilament line trailed from its cloaca, and radiography revealed that the turtle had ingested two additional hooks and a large sinker. The hemogram showed leukocytosis. At exploratory celiotomy, the fishing line was seen to have acted as a linear foreign body and had perforated the intestines. Multiple enterotomies were performed to remove the sinker and line, and perforations were repaired. Two of the hooks could not be surgically or endoscopically retrieved. Blood lead concentration was 3.6 ppm prior to start of chelation therapy with calcium disodium ethylenediaminetetraacetic acid and declined to undetectable levels within 6 wk. The turtle recovered and was released. PMID- 9226627 TI - Clinical challenge. Osteomyelitis of the distal half of the metacarpus with associated involucrum and bone sequestrum formation. PMID- 9226629 TI - Separation of multiple yellow fluorescent lipofuscin components in rat kidney and their characterization. AB - Yellow fluorescent lipofuscin deposited in rat kidney was extracted in an aqueous solution and characterized after separation. Centrifugal fractionation of the extract revealed that most of the yellow fluorescence was detected in the 105,000 x g-supernatant, and little in nuclei, cell debris, mitochondria, lysosomes, microsomes and plasma membrane. The yellow fluorescence in the supernatant was fractionated by gel filtration through Sephadex columns into 5 yellow fluorescent fractions A, B (B1, B2 and B3) and C showing the same fluorescence spectra with excitation maximum/emission maximum at 400/620 nm. The components in fraction A were converted into the smaller molecular-weight components in fraction B on treatment with 4 M urea or protease, suggesting that they were proteinaceous. The smallest molecular-weight fluorescent components in fraction C were adherent to solid cellulose materials. The fluorescent components in all the fractions were soluble in water and insoluble in chloroform-methanol, indicating that they were not lipidic materials. The fluorophores in these fractions were kept stable on borohydride treatment, but readily converted into non-fluorescent components on heavy-metal ion treatment. The characteristics of the yellow fluorescence in these fractions were quite different from those of bluish lipofuscin-like fluorophores that may be generated in tissues during lipid peroxidation. PMID- 9226628 TI - Effect of melatonin and pineal peptide preparation epithalamin on life span and free radical oxidation in Drosophila melanogaster. AB - It was shown previously that epithalamin delays age-related changes in reproductive and immune systems and increases the life span of mice and rats. These effects could be mediated by stimulating influences of epithalamin on synthesis and secretion of melatonin and on free radical processes. A comparative study on the effect of epithalamin and melatonin on both the life span of Drosophila melanogaster (strain HEM) and on the intensity of lipid peroxidation and activity of antioxidative enzymes in their tissues was the main aim of this work. Melatonin and epithalamin was added to the nutrition medium (100 micrograms/ml) during 2-3rd age of larvas. For survival analysis the flies were passed (five coupes per vessel) each 3-7 days. Lipid peroxidation was evaluated as the level of ketodienes (KD) and conjugated hydroperoxides (CHP) in fly tissues at the age of 11 days. Activity of Cu, Zn-superoxide dismuatse (SOD) and catalase was evaluated as well. The mean, median and maximum life span (MLS) were estimated. Mortality rate (MR) was calculated as alpha in the Gompertz equation (R = Ro (exp alpha t) and mortality rate doubling time (MRDT) as in 2/alpha. These parameters in groups of male and female flies exposed to melatonin and in male flies exposed to epithalamin were no different from the parameters for controls. However, exposure to epithalamin was followed in females by a significant increase in mean life span (by 17%, P < 0.02), of median (by 26%), of MLS by 14% and by a 2.12 times decrease of MR (P < 0.01) and MRDT (by 32%) compared with female controls. The level of CHP and KD in the tissues of male control flies was 40 and 49% less than that in females and indirectly correlates with male life span. Exposure to melatonin was followed by a decrease in the level of CHP and KD in females and the deletion of sex differences in them. Exposure to epithalamin significantly decreased the level of CHP and KD in female flies compared to controls (2.3 and 3.4 times, respectively, P < 0.001). Exposure to melatonin failed to influence the activity of catalase in males but increased it in females by 24% (P < 0.02) and failed to influence SOD activity both in males and females. Exposure to epithalamin was followed by a significant increase in activity of catalse, 20% in males and 7% in females and by an increase in SOD activity in males (41%). Thus, it was shown that exposure to epithalamin significantly increases the mean life span and MLS of female D.melanogaster and slowed down their aging rate by 2.12 times. This effect is in good agreement with the inhibiting effect of epithalamin in lipid peroxidation processes in fly tissues. PMID- 9226630 TI - In K562 and HL60 cells membrane ageing during cell growth is associated with changes in cholesterol concentration. AB - In a cell culture model of aging we have previously shown that there is an age related decrease in the lipid dynamics of the proerythropoetic K562 cell membranes, as determined by the generalized polarization (GP) of the phase sensitive lipid probe 2-dimethylamino-6-lauroylnaphthalene (Laurdan) (T. Parasassi, M. Di Stefano, G. Ravagnan, O. Sapora and E. Gratton. Exp. Cell Res., 202 (1992) 432-439). In the present study we also extended our observations to the lymphoblastoid HL60 cell line. In both K562 and HL60 cells during the four days after the last cell culture medium renewal the GP Laurdan value increased in a linear fashion indicating a time-dependent decrease in lipid dynamics. The initial membrane physical properties were almost completely restored upon renewal of the cell culture medium. We measured lipid composition, including individual and total phospholipids, free and esterified cholesterol at the first ('young') and at the fourth ('aged') day after culture medium renewal. We found that the decreased membrane lipid 'fluidity' at the fourth day of cell growth was associated with a 40% increase in cholesterol concentration in both cell lines. This increase in cholesterol concentration was reversible 24 h following the culture medium change. We conclude that in K562 and HL60 cells the 'age-related' decrease in membrane lipid dynamics is mediated by an 'age-related' increase in cell cholesterol content. PMID- 9226631 TI - The effects of age on rabbit MCL fibroblast matrix synthesis in response to TGF beta 1 or EGF. AB - In this study, we examined the effects of age on collagen and total protein synthesis by ligament fibroblasts in response to growth factors. Three different doses of transforming growth factor-beta 1 (TGF-beta 1) or epidermal growth factor (EGF) were individually added to in vitro fibroblast cultures from the medial collateral ligament (MCL) of skeletally immature (age 3 months), mature (age 12 months) and senescent (age 48-51 months) rabbits. Analysis of the effects of age revealed that fibroblasts from senescent rabbits produced significantly less collagen in response to TGF-beta 1 or EGF stimulation when compared to fibroblasts from immature rabbits. Furthermore, increased age was found to result in significant reductions in the baseline levels of collagen synthesis but not total protein synthesis. Additionally, collagen and total protein synthesis by MCL fibroblasts were significantly affected by the TFG-beta 1 dose, but not by the EGF dose. When fibroblasts were normalized to their own controls, the increase in collagen and total protein synthesis due to TGF-beta 1 and EGF for the senescent group were found to be greater than those for the skeletally immature rabbits at all doses. This demonstrates that MCL fibroblasts from senescent rabbits are responsive to growth factors. PMID- 9226633 TI - The magnocellular neurosecretory system of the hamster hypothalamus: an ultrastructural and morphometric study during lifetime. AB - A quantitative study regarding the age-related changes occurring in the nucleus and the somatic organelles of neurosecretory magnocellular neurons of the hypothalamo neurohypophyseal system (HNS) was carried out in the hamster at six age-points during animal life. The magnocellular cells of both parts of the supraoptic nucleus (SON) and the paraventricular nucleus (PVN) of male Syrian hamsters between 3 and 30 months of age were examined ultrastructurally. Cells of all age groups present the same morphological ultrastructure. Standard manual morphometric techniques are used to calculate the following parameters related directly or indirectly with cellular activity: nuclear area, nucleolar area, nuclear invagination index and volumetric fractions of some intracellular structures (Golgi apparatus, mitochondria, rough endoplasmic reticulum and lipofuscin). With respect to the cell nucleus, the parameters are not modified during aging. No significant differences in the volume density of subcellular components, except lipofuscin, were detected at the age groups studies. However, there is a positive linear trend among all parameters and age except for the rough endoplasmic reticulum. Our results suggest maintenance of the synthetic activity of the magnocellular neurons in the hamster during aging but in no case an increase in their metabolic activity. PMID- 9226632 TI - Abundance of alpha 1(I) and alpha 1(III) procollagen and p21 mRNAs in fibroblasts cultured from fetal and postnatal dermis. AB - The steady-state abundance of alpha 1(I) and alpha 1(III) procollagen mRNAs, p21Sdi1 mRNA, and beta-actin mRNA was determined in 29 skin fibroblast lines established from fetal, young and old donors. Donor ages ranged from 12 gestational weeks to nonagenarian. Adult donors were members of the Baltimore Longitudinal Study of Aging. The abundance of alpha 1(I) procollagen mRNA was decreased in cell lines from both young and old donors compared with fetal lines. Additionally, one alpha 1(I) transcript observed in the fetal lines was not detected in postnatal lines. The abundance of alpha 1(III) procollagen mRNA was decreased in postnatal lines from old donors compared with fetal lines. The abundance of beta-actin mRNA was lower in postnatal lines from both young and old donors compared to fetal lines. These results suggest that cultures of fetal skin fibroblasts exhibit a greater capacity for synthesis of procollagens and beta actin than postnatal lines. In contrast, the abundance of p21Sdi1 mRNA was elevated in lines established from postnatal donors. These results are consistent with developmental changes in amounts of procollagen, beta-actin and p21. PMID- 9226634 TI - Plasma renin activity and metabolic clearance rate of angiotensin II in the unstressed aging rat. AB - We conducted studies in conscious chronically catheterized, trained young (3-5 months) and old (18-20 months) rats to assess the impact of aging on baseline renin activity (PRA) and metabolic clearance rate (MCR) of angiotensin II (ANG II). We observed that under unstressed conditions the baseline values of PRA and plasma ANG II were no different in young versus old rats (1.8 +/- 0.2 versus 1.5 +/- 0.2 ng Al/ml/h and 18 +/- 3 versus 15 +/- 2 fmol/ml, respectively). Values of PRA in the present study were similar to those reported by others for old rats, but our young rat values were lower than usually reported. This probably reflects our use of an unstressed preparation. We also observed a blunted increase in PRA in old rats in response to acute converting enzyme inhibition. Overall, our observations suggest that old rats may lose their ability to increase PRA in response to acute stimuli, including perhaps, the stress of blood drawing in emotionally or surgically stressed preparations. We also observed that the MCR of ANG II increased with age, despite similar baseline plasma ANG II concentrations in young and old. This suggests that with aging, an increase occurs in the rate of synthesis of ANG II. These results emphasize the importance of establishing true baseline values for indices of the renin-ANG II system in aging. PMID- 9226635 TI - NOR activity in hippocampal areas during the postnatal development and ageing. AB - The silver staining of the nucleolar organizer regions (Ag-NORs) was used in order to estimate the biosynthetic activity of three hippocampal areas (dentate gyrus, CA1 and CA3) during postnatal development and ageing. 32 Wistar rats were used and 4 groups were formed according to the age of the animals (14, 21, 90 days and 23 months). Several Ag-NOR parameters such as mean Ag-NOR area and the ratio between Ag-NOR and nuclear areas per neuronal cell were quantified using an image analysis system. High values of these parameters are associated with a high rate of rRNA transcription. In this way, the neural biosynthetic activity in all regions studied decreased as the older ages are reached. Differences between areas are shown with the dentate gyrus and CA1 areas decreasing faster. The different activity among these areas is discussed, taking into account the particular affect on these areas of some injuries and the ageing process. Our results support the hypothesis of NOR loss as a main cause of ageing as reported by other authors. PMID- 9226636 TI - Sensitivity of the segmental renal arterioles to angiotensin II in the aging rat. AB - With advancing age the old rat kidney becomes tonically vasoconstricted by endogenous angiotensin II (ANGII) (C. Baylis. Am. J. Kid. Dis., (1993) 842). The present study was designed to investigate the sensitivity of the cortical glomerular microvasculature of the old rat kidney (19-22 months of age) to exogenous ANGII, using the in vivo micropuncture technique. In the baseline state, glomerular blood pressure (P(GC)) in old male rate was higher compared to young rats (4-5 months of age). During exogenous ANGII infusion (40 ng/kg/min), a significant rise in arterial blood pressure and renal vasoconstriction occurred in both young and old rats. In young rats, the ANGII induced fall in renal plasma flow (RPF) and glomerular plasma flow (QA) was accompanied by a rise in PGC and thus the glomerular hydrostatic pressure gradient, with little change in Kf. Therefore, the glomerular filtration rate (GFR) and single nephron GFR (SNGFR) were unchanged by ANGII infusion in young rate. In old rats, RPF and QA fell, a rise occurred in PGC and also a fall was seen in the glomerular capillary ultrafiltration coefficient (Kf), thus GFR and SNGFR fell significantly. The magnitude of the pressor and renal vasoconstriction response to ANGII were not affected by age; of interest, ANGII increased preglomerular and efferent arteriolar resistance (RA, RE) and PGC by similar accounts in young and old rats. SNGFR was reduced in old rats, due to the marked ANGII-induced decline in Kf. Neither absolute nor fractional proximal reabsorbtion were affected by ANGII infusion in either young or old rats. by 19-22 months of age, old rats had much more injured glomeruli than young rats. These data demonstrate that the afferent and efferent arterioles had similar sensitivity to exogenous pressor dose of ANGII in both young and old rats, but Kf was more sensitive to ANGII in old rats leading to a significant fall in SNGFR. PMID- 9226637 TI - Protection of dogs from bites of phlebotomine sandflies by deltamethrin collars for control of canine leishmaniasis. AB - Dog collars made of PVC plastic impregnated with the pyrethroid insecticide deltamethrin at 40 mg/g were investigated for their protective efficacy against phlebotomine sandflies. Collared dogs were kept separately (two untreated control dogs lived together) in outdoor enclosures, each with a kennel, in the Cevennes, southern France. To measure sandfly mortality and anti-feeding effects due to the deltamethrin-impregnated collars worn continuously by the dogs for up to 8 months, each dog was periodically sedated and exposed for 2h to 150-200 laboratory-reared Phlebotomus perniciosus females (plus c. 25 males) inside a net (1.2 m square, 1.8 m high) indoors. After dogs were removed from the nets, allowed to recover and returned to their kennels, any dead sandflies were collected from inside the net and counted. Surviving flies were kept overnight, then scored according to whether they were still alive or dead, unfed or blood fed. From tests 2, 3, 4, 13, 20, 26 and 34 weeks after the dogs began wearing collars, the overall numbers of blood-fed female sandflies recaptured were 75 from two dogs with collars, compared with 1911 from two collarless dogs. Thus, for every 100 flies which fed on collarless dogs, only 4 fed on collared dogs, i.e. the collars protected dogs from 96% of the bites and this activity was maintained for up to 34 weeks. During the same period, the percentage of recaptured female sandflies that had fed on collared dogs was 0-12% compared to 55-95% on collarless dogs. Immediately after dogs were taken out of the nets, 21 60% of flies confined with the collared dogs were found dead, compared to 0-12% with the controls. It is concluded that, at least in the Mediterranean subregion, this insecticidal collar would protect a dog from the majority of sandfly bites and retain a killing effect for a complete sandfly season. Moreover, it seems likely that the use of collars on all dogs in a focus of Leishmania infantum would reduce contact between sandfly vectors and canine reservoir hosts sufficiently to diminish the risk of infection for humans as well as dogs. PMID- 9226638 TI - Larval and adult nutrition effects on blood/nectar choice of Culex nigripalpus mosquitoes. AB - The impact of nutritional variables on the development of host-seeking and biting behaviours after emergence by female Culex nigripalpus mosquitoes were studied using air-flow olfactometer and close-range biting assays, respectively. Unfed females failed to develop resting stage ovarian follicles. When offered a bird host in the absence of competing stimuli, sugar-fed mosquitoes were significantly more responsive in both host-seeking and biting than unfed controls. In a choice olfactometer assay using nectar odours (honey scented with artificial apple blossom oil) versus host odours (a bird), unfed females preferred honey over bird odours except when honey odour was weak. After sucrose feeding, females switched from honey to bird preference. This change in behaviour was accompanied by significant accumulation of lipid and by follicular growth to the resting stage. Elevation of host responsiveness after sugar feeding was reversible; starvation ultimately resulted in females preferring honey over bird odours. When the larval diet was restricted by crowding, the wing-length and total lipid of resultant adult females were reduced. Although differences were subtle, unfed bird responding females tended to have longer wings and more lipid than their honey responding counterparts. PMID- 9226640 TI - Chorion patterns on eggs of Lutzomyia sandflies from the Peruvian Andes. AB - Scanning electron microscopy (SEM) as used to depict and describe the eggs of five species of phlebotomine sandfly from the Andean region of Peru: Lutzomyia caballeroi, Lu.noguchii, Lu.peruensis, Lu.tejadai and Lu.verrucarum. Two new types of chorionic sculpturing of sandfly eggs are reported: these were named disperse (Lu.tejadai) and verrucose (Lu.noguchii). The aeropylar area of the eggs is also described for the first time for neotropical sandflies. These character states appear to vary for ecological rather than phylogenetic reasons and could be used for species identification. PMID- 9226639 TI - Esterases A5-B5 in organophosphate-resistant Culex pipiens from Italy. AB - Culex pipiens mosquitos from Lignano city, Udine province, northeast Italy, were found to carry over-produced non-specific esterases A1, A2-B2 and A4-B4 or A5-B5, detected by starch gel electrophoresis, giving multiple resistance to organophosphorus insecticides. In order to differentiate between A4-B4 and A5-B5 esterases, the latter known only from Cyprus whereas the former is widespread in Italy and elsewhere, restriction fragment length polymorphism (RFLP) analysis was performed at the esterase B locus. Both B4 and B5 haplotypes were found. This is the first record of A5-B5 esterase-mediated resistance in continental Europe. PMID- 9226641 TI - Ribosomal DNA difference between species A and D of the Anopheles dirus complex of mosquitoes from China. AB - Species A and D of the Anopheles dirus complex were found in China. Ribosomal DNA second internal transcribed spacers (ITS2) of both species A and D were sequenced and found to be 716 and 710 base-pairs in length, respectively, with 69% GC content. No evidence of intraspecific variation was detected in the ITS2 sequence of species A, whereas the sequence of species D showed variation at one position in the ITS2. A large number of simple repeat motifs were dispersed throughout the ITS2 sequences. The level of interspecific difference was 5.4% of the nucleotide sequences. Some of the interspecific differences were located in regions with subrepeat structure. PMID- 9226642 TI - Preliminary investigations of Aphodius species activity in cattle faeces treated with ivermectin. AB - A.sphacelatus at densities of 0.5 and 1.0 beetle/g faeces caused significantly greater median percentage reductions (65.2% and 87.4% respectively) of Pilobolus sporangia than 0.1 beetle/g faeces (31%) in untreated cattle faeces. The median percentage reduction in sporangia due to beetle activity (48.4%) was significantly lower (P < 0.02) in faeces mixed with ivermectin at 1.0 ppm (wet weight) than in untreated faeces (88.4%). After treating a bullock with ivermectin (IvomecR Pour-On), the median percentage reduction in sporangia caused by beetles was significantly less (P < 0.05) on days 9 (78.9%) and 10 (76.9%) than in pre-dose faeces (86.5% and 93.8% respectively). In microcosms without beetles, sporulation of Pilobolus in cattle faeces from a heifer treated with ivermectin was significantly less on days 5, 10 and 15 after dosing. However, this difference was not apparent for days 5 and 10 after storage of faeces at 4 degrees C for 55 and 50 days respectively. PMID- 9226643 TI - The effects of the bont tick, Amblyomma hebraeum, on milk production of Sanga and Sanga x Brahman cattle. AB - The effects of adults on the bont tick, Amblyomma hebraeum on the milk production of Sanga and Sanga x zebu (Brahman) cattle were measured over a period of 11 weeks in the low veld of Zimbabwe in the summer of 1986. Four groups of lactating cows, consisting of two breeds, each divided into a high and low tick treatment, were exposed to very low or high challenges of ticks and their milk production measured by weighing their calves before and after suckling. The liveweight gains (LWG) of the calves were also measured. Tick burdens on the infested groups averaged around fourteen engorging females of A.hebraeum per day, which amounted to infestations of about 150 adult ticks. This is greater than most observed field infestations. This caused no significant reduction in milk yield or calf growth over the whole period, provided the teats of the dams had not been damaged by ticks. Mismothering occurred when teats were damaged. No breed differences were observed so all data was pooled for further analysis. Average calf LWGs of the high tick groups were reduced by 2.2 kg (P < 0.01) during one 4-week period but overall the 3.9 kg difference in LWG of the tick treatment groups was not quite significant (P < 0.10). Although there was a poor relationship between tick numbers and reduced milk yield or calf LWG, the effects were always in the direction expected. The effects averaged 6 +/- 10 g reduction of milk and 2.6 +/- 1.8 g loss of LWG of calves for every female tick that engorged. it was concluded that milk production is not an important consideration when estimating the losses in production caused by A.hebraeum on Brahman x Sanga or Sanga breeds of cattle. Losses due to teat or udder damage could be much more important and need to be quantified. PMID- 9226644 TI - The effects of the brown ear-tick, Rhipicephalus appendiculatus, on milk production of Sanga cattle. AB - Lactating Sanga cows of the Mashona breed from Zimbabwe, receiving either a low or high level of nutritional supplement, were exposed to two levels of infestation of adults of the brown ear-tick, Rhipicephalus appendiculatus in the highveld of Zimbabwe. The effect of the ticks on the milk yield was measured over an 11-week period during the rainy season from January to April 1986. A technique in which calves were weighed before and after suckling was used to estimate milk yield. There were significant treatment effects of ticks (P < 0.05) on milk production but no significant differences in liveweight gain between calves from tick-free and tick-infested dams. The loss in milk production was poorly related to the number of female ticks that engorged, being 9 g (SEM 4) per tick. A Friesian x Hereford (Bos taurus) reference group of cattle carried 50% more ticks than the Mashona cows, illustrating a difference in resistance between the breeds. Thirteen screw-worm (Chrysomya bezziana) strikes were recorded amongst the thirty-two Mashona cows compared with twenty-one amongst the ten Friesian x Hereford animals between January and the end of March. PMID- 9226645 TI - The effects of the brown ear-tick, Rhipicephalus appendiculatus, on milk production in dairy cattle. AB - The effect is reported of artificially controlled levels of infestation with adults of Rhipicephalus appendiculatus on the milk yield of twenty commercial Bos taurus dairy cattle on a high plane of nutrition and eighteen crossbred B.taurus x Sanga cattle on a lower plane of nutrition on the highveld of Zimbabwe. The results showed no significant effect on milk yield of infestations averaging twenty engorging ticks per animal per day, despite severe ear damage in some animals. They indicate that milk production of dairy cattle under commercial management is not sensitive to infestation with R.appendiculatus. The results have important implications for management of ticks in Africa, but need to be interpreted within the context of the control of tick-borne diseases. PMID- 9226646 TI - Observations on the mite fauna associated with adult Stomoxys calcitrans in the U.K. AB - Adult females of the blood-sucking muscid Stomoxys calcitrans sampled between June and September 1993 from a cattle farm (n = 839) and from a pig farm (n = 542) in North-West England were examined for mites. Twelve species of mites from ten families and three orders were identified as follows. In the Prostigmata, Eryenetes sp., Family Ereynetidae and Pediculaster mesembrinae, Family Pygmephoridae. In the Astigmata, Procalvolia zacheri Family Saproglyphidae, Acarus farris, Family Acaridae, Bonomoia sphaerocerae and Myianoetus sp., Family Anoetidae. In the Mesostigmata, Macrocheles muscaedomesticae and Macrocheles subbadius Family Macrochelidae, Digamasellus sp., Family Digamasellidae, Halolaelaps sp., Family Halolaelapidae. Prodinychus sp., Family Uropodoidea and Thinoseius sp., Family Eviphididae. Mean infestation rates at the two sites (all mite species) for the entire sampling period were 31.6 +/- 13.9% and 19.8 +/- 3.6% respectively. 51% of synbovine flies sampled in July were infested with mites. Mean numbers of flies infested in August at both farms were significantly lower compared to other months. The presence of tritonymphs of Ereynetes sp. on S. calcitrans demonstrates for the first time that this life cycle stage is naturally associated with insects in the field. All mites were recovered from the ventral thorax and abdomen, and two or more species commonly infested individual flies. Associations of mites with their dipteran hosts are described and discussed. PMID- 9226647 TI - Scanning electron microscopy studies on the first-instar larva of Dermatobia hominis. AB - First-instar larvae of Dermatobia hominis collected 1, 4 and 7 days after having penetrated experimentally infected rats, were studied by scanning electron microscope (SEM) observation. On the pseudocephalon there are basiconic and trichoid sensilla (antennal sensory complex), and basiconic, coeloconic and campaniform sensilla (maxillary sensory complex). The thoracic segments bear several rows of small, backwardly pointed, spines, and trichoid, campaniform, coeloconic and pit sensilla. The anterior spiracle is a minute opening. Both small and large spines directed posteriorly are on the first to fourth abdominal segments, which also bear coeloconic and companiform sensilla. These sensilla are present on the unarmed (fifth and sixth) and armed (seventh) abdominal segments. The seventh and the last (eight) abdominal segments have forwardly directed spines. Each spiracular plate has two spiracular openings and four spatulate-like structures called sun rays. The anus and the coeloconic sensilla are proeminent on the last segment. The results are compared with other parasitic dipteran larvae, and emphasize that the multiple types of sensilla on D.hominis larva may have importance in establishing the parasitic phase of the life cycle of this insect. PMID- 9226648 TI - Transmission of louping ill virus between infected and uninfected ticks co feeding on mountain hares. AB - Most of the data on oral infection of ticks by louping ill virus have been obtained from experiments in which animals were infected by syringe inoculation with infectious material. Using infected ticks to mimic the natural situation, we have demonstrated that louping ill (LI) virus transmission can occur from infected to uninfected Ixodes ricinus feeding in close proximity on mountain hares (Lepus timidus). Under these conditions the hares developed either low or undetectable viraemias. Highest prevalence of LI virus infection was observed in recipient nymphs which had fed to repletion between days 3 and 7 post-attachment of virus-infected adults; following engorgement, 56% of nymphs acquired virus. These results demonstrate the efficient transmission of LI virus between co feeding ticks on naive mountain hares. However, when ticks were allowed to co feed on virus-immune hares a significant reduction in the frequency of infection was observed. Neither red deer (Cervus elaphus) nor New Zealand White rabbits supported transmission of LI virus. The significance of virus transmission between cofeeding ticks on LI virus epidemiology is discussed. PMID- 9226649 TI - Responses of mosquitoes of the Anopheles farauti complex to 1-octen-3-ol and light in combination with carbon dioxide in northern Queensland, Australia. AB - In northern Queensland, Australia, three experiments were conducted to determine the response of mosquitoes of the Anopheles farauti complex to CDC traps baited with four attractant combinations: octenol + CO2 and light; octenol and light; CO2 and light; or CO2 and octenol without light. A CDC-modified updraft light trap was also trialled, but did not significantly enhance collections of An.farauti sensu lato. The combination of light, octenol and CO2 caught significantly more An.farauti s.l. (both An.farauti No.1 and No.2 sibling species) when compared to CO2 and light alone. Only small numbers of the An.farauti complex were captured when CDC traps were baited with octenol alone, i.e. no light or CO2. PMID- 9226650 TI - Development and survival of immature Culex annulirostris mosquitoes in southeast Queensland. AB - The rates of development and survival of the immature stages of Culex annulirostris were studied in three different breeding sites in the Brisbane area of southeast Queensland: a temporary rain-filled pool (TP), a semi-permanent pool (SPP), and an area of flooded grassland (FG) arising from the overflow of a permanent pond. Parallel observations were made on larvae and pupae either exposed to predation or in predator-free cages of 12 and 15 consecutive days respectively. Development was fastest and survival highest in the warmer TP. Average daily mortality of larvae (L) and pupae (P) was TP: L 8.3%, P 9.5%; SPP: L 25.2%, P 34.6%; FG: L 18.8%, P 35.4%. From life-tables and survivorship curves, survival from first instar to adult was TP 34.8%, SPP 2.0% and FG 4.2%. Mortality from predation was TP 43.2%, SPP 68.7% and FG 69.1%. Predator density was TP 0.32, SPP 1.48 and FG 1.76 per 300 ml sample. PMID- 9226651 TI - Summer mastitis experimentally induced by Hydrotaea irritans exposed to bacteria. AB - Summer mastitis is an acute suppurative bacterial infection of the udder in heifers and dry cows. To ascertain the possible role of flies in the transmission of the disease, experimental exposures of recipient heifers to Hydrotaea irritans previously exposed to bacteria were carried out. Flies were allowed to feed on secretions from clinical cases of summer mastitis. The pathogens present were the bacteria Staphylococcus aureus, Streptococcus dysgalactiae, Actinomyces pyogenes, Stuart-Schwan cocci, Peptococcus indolicus, Fusobacterium necrophorum and Bacterioides species. The teats of eight heifers were exposed to flies with verified pathogen content. Two teats of each animal were deliberately damaged before fly exposure. One teat was cut, another pricked with insect needles to mimic insect bites. Two of the heifers developed summer mastitis in the quarters where teats had been cut. The bacterial species isolated from these quarters corresponded to those that had previously been fed to the flies. For the first time, it is now demonstrated that H.irritans is capable of transmitting summer mastitis pathogens and so causing summer mastitis in recipient heifers. Lesions on the teat orifice may be a predisposing factor in the development of the disease. PMID- 9226652 TI - Gonotrophic and embryonic development of the Palaearctic screwworm fly Wohlfahrtia magnifica. AB - Gonotrophic and embryonic development of Wohlfahrtia magnifica were studied on pupae and adults reared in the laboratory. Gonotrophic development in screwworm begins 8 days after pupation in yellow eye pharate adults. The adult female emerges with its oocytes at stage 3 and requires only 2 days for complete oogenesis and 5 days for complete embryogenesis. Mating occurs 2-5 days after adult emergence, starting the embryonic development. Each female produces an average of 76.8 active larvae which can produce a traumatic lesion in their hosts. When food sources were regularly supplied synchronization of gonotrophic stages can be observed. These stages together with the embryonic ones are described. PMID- 9226653 TI - Anti-OspA antibody reduces reservoir competence of mice for Borrelia burgdorferi, the agent of Lyme disease. PMID- 9226654 TI - Effect of infestation with Rhipicephalus sanguineus on the antibody productivity in dogs. PMID- 9226655 TI - Visual detection in monkeys with blindsight. AB - Monkeys with unilateral striate cortical removal show residual visual abilities in their affected hemifield. To learn whether the monkeys, like patients with blindsight, lose the phenomenal representation of the visual stimuli they nevertheless respond to, we first studied their ability to localize a briefly presented target in either hemifield. By varying the luminance of the stimuli we determined their visual sensitivity, which was reduced by 0.3-1.5 log units in the impaired hemifield; suprathreshold stimuli yielded almost perfect performance. We then presented two tests designed to show whether the monkeys categorized visual stimuli in the impaired field in the same manner as they categorize them in the normal field. In the first test, they had to respond differently according to whether one or two lights were presented, with the relative position of the two stimuli in a pair being varied. Whenever one of the paired stimuli lay in the impaired hemifield, two of the three monkeys consistently ignored it, and responded as if it had been a single stimulus in the good field. In the second test, trials consisting of a single stimulus light were interleaved with blank trials. The monkey touched the position of the light or made a different response, indicating that no light had appeared. All three monkeys responded to a light of supra-threshold luminance presented in the impaired field as if it were a blank trial. These results suggest that monkeys with striate cortical destruction, like neurological patients with similar lesions, have blindsight rather than phenomenal vision when they have to detect brief static visual targets. PMID- 9226656 TI - Rightward attentional bias and left hemisphere dominance in a cue-target light detection task in a callosotomy patient. AB - Six normal subjects and a callosotomized man with a prefrontal lesion, mostly on the right side, were tested in a reaction time (RT) task involving a key-pressing response to an extrafoveal light target preceded by an extrafoveal light cue. Cues and targets were presented along the horizontal meridian at 4 degrees and 12 degrees on the right and left of fixation. Fixation was maintained throughout each trial. The cue signalled the occurrence of the target within a time window extending from 200 to 4000 misec from the cue, but did not predict target location. Normal controls responded faster to medial than to lateral targets in both fields, but showed no between-field difference, and their RT was not affected by cue location. Furthermore, they showed the so-called 'ipsilateral inhibition' or 'inhibition of return' effect, their RT being longer when cues and targets occurred in the same field than when they occurred in opposite fields. The RT of the callosotomized subject showed a left-right gradient for both cue location and target location, being longest for the leftmost location and shortest for the right locations. In addition, he showed a significant advantage for the right hand regardless of cue and target location, as well as a consistent ipsilateral inhibition in the left field, whereas in the right field there was ipsilateral inhibition only at the two longest stimulus onset asynchronies. These results suggest that, at least under these experimental conditions, there was a rightward orientational bias which reflected the taking over of the control of performance by the left hemisphere. This attentional bias was reminiscent of that seen in patients with hemi-inattention from right hemisphere damage, although the callosotomized patient showed no sign of such hemi-inattention in routine clinical tests. On the basis of several considerations the rightward bias could be attributed to the callosal interhemispheric disconnection rather than to the right prefrontal lesion. PMID- 9226657 TI - Effects of lesions invading the postcentral gyrus on somatosensory thresholds on the face. AB - Patients with unilateral removals from either the parietal, frontal or temporal lobe and normal control subjects were examined on three tests of tactile sensibility. The patients with surgical excisions from the parietal lobe were subdivided into two groups: those whose lesions invaded the face area in the primary sensory cortex and those whose lesions spared this area. A significant percentage of patients with lesions that invaded the face area had mild to severe sensory deficits on the side of the face contralateral to the lesion. A much smaller number of patients had deficits on the ipsilateral side. Lesions to the face area in the primary sensory cortex were, however, associated with a lower incidence of severe and persistent sensory deficits when compared to previous results on the effects of lesions to the hand area on the sensory capacity of the hand. These results suggest that there is some preservation of sensory function after damage to the face area in the primary sensory cortex, presumably due to the bilateral representation of the face. PMID- 9226658 TI - Learning and retention of words and designs following excision from medial or lateral temporal-lobe structures. AB - We sought to elucidate the contributions of the amygdala, hippocampus and temporal neocortex to learning and memory for verbal and visuospatial material. Two matched learning tasks, using abstract words versus abstract designs, were administered to patients with unilateral neocorticectomy (NCE; Dublin), selective amygdalohippocampectomy (AHE; Zurich) or anterior temporal-lobe resection invading the amygdala and hippocampus (ATL; Montreal). Data were analysed according to side and type of resection. Learning and recall for words was impaired in groups with resection from the left temporal lobe, irrespective of whether mediobasal structures were spared or temporal neocortex was spared. All right-resection groups were unimpaired. Learning for abstract designs was impaired across all trials in the right AHE and NCE groups, and on the last two trials in the right ATL group. Restricted deficits of lower magnitude were observed on some trials in left-resection groups. These results show a partial dissociation between side of excision and type of material, but the finding of similar deficits in all resection types was unexpected. We propose that excision from either the hippocampal region or temporal neocortex may result in a disconnection, giving a similar functional outcome, as both types of resection interrupt a circuit likely to be essential for normal storage and retrieval of information. PMID- 9226659 TI - Severe amnesia: an usual late complication after temporal lobectomy. AB - A patient developed the severe amnesic syndrome 8 years after temporal lobe surgery for epilepsy. He underwent left temporal lobectomy (6 cm, 43.5 g; hippocampal sclerosis) aged 19, and remained seizure free for 8 years until a convulsion followed a head injury. He became severely amnesic after a fourth convulsion 16 months later. He was right-handed, pre-operative IQ was average, verbal memory poor and non-verbal memory normal. Post-operatively, these were unchanged. After the first post-operative seizure he began professional training. After onset of amnesia IQ was unchanged, anterograde memory severely impaired and retrograde amnesia dense for at least 16 months. He died 2 years later. Magnetic resonance imaging before amnesia showed absence of anterior left temporal lobe, atrophy of left fornix and mamillary body, and normal right temporal lobe. Four months after onset of amnesia, right hippocampal volume had reduced by 36%. Autopsy showed: previous left temporal lobectomy with absence of left amygdala and hippocampus, atrophy of fornix and mamillary body; neuronal loss in the right hippocampus, severe in CA1 and CA4; intact right amygdala and parahippocampal gyrus; recent diffuse damage associated with cause of death. A convulsion can cause severe hippocampal damage in adult life. Hippocampal zones CA1 and/or CA4 are critical for maintaining memory and the amygdala and parahippocampal gyrus cortex alone cannot support acquisition of new memories. PMID- 9226660 TI - Visuo-motor conditional associative learning after frontal and temporal lesions in the human brain. AB - It has been shown that damage to the human lateral frontal cortex results in a severe impairment on conditional associative tasks requiring learning of arbitrary associations between a set of stimuli and a set of responses (Petrides, M., Neuropsychologia, 1985, 23, 601-614; 1990, 28, 137-149). In these studies, which first demonstrated the impairment after frontal lesions, training was by a trial-and-error procedure, during which the subject performed the various responses when a given stimulus was presented and the experimenter provided feedback until the correct response was performed. In the present experiment, patients with unilateral frontal- or temporal-lobe excisions were tested on a visuo-motor conditional associative task with a modified procedure. The subjects had to learn arbitrary associations between a set of coloured stimuli and a set of hand postures. Training in the present experiment consisted of a series of demonstration trials followed by test trials. In the demonstration trials, the experimenter showed the subject the associations between the stimuli and the responses and, in the test trials that followed, the subject was tested on these associations. If an error was made on the test trials, the correct response was demonstrated by the experimenter. Despite these changes in the training procedure, namely the demonstration of the stimulus-response associations and the provision of the correct response immediately following an error, patients with left or right frontal-lobe excisions were severely impaired in learning this task. These findings, together with those of the earlier studies (Petrides, M., Neuropsychologia, 1985, 23, 601-614; 1990, 28, 137-149), demonstrate that the impairment in conditional learning after frontal lesions is not dependent on the type of the training procedure and therefore that it reflects a specific impairment in learning. PMID- 9226661 TI - Effects of orbital frontal and anterior cingulate lesions on object and spatial memory in rhesus monkeys. AB - Object memory processes, evaluated in rhesus monkeys by delayed nonmatching-to sample with trial-unique stimuli and object reversal learning, were more severely impaired by orbital frontal than by anterior cingulate lesions. Spatial memory processes, assessed by spatial delayed response and spatial reversal learning, showed a weak trend in the opposite direction, though on these tasks neither lesion produced a serious loss. Comparison of the present results with those of earlier studies on the effects of various limbic system lesions suggests that object memory processes, including object recognition and object-reward association, are served by a circuit consisting mainly of the rhinal cortex, orbitofrontal cortex, and the magnocellular division of the medial dorsal thalamic nucleus. Although both the rhinal and orbitofrontal components of this circuit appear to participate in both functions, evidence from the present and earlier studies suggests that the orbitofrontal component is the more important one for associative memory, i.e. the formation across trials of associations between particular objects or classes of objects and reward, whereas the rhinal component is the more critical one for recognition memory, i.e. the storage and retrieval within trials of the representations of particular objects. PMID- 9226662 TI - Strategic retrieval and the frontal lobes: evidence from confabulation and amnesia. AB - Confabulation and amnesia are considered disorders of episodic but not of semantic memory. To test the limits of this view, retrieval from episodic and semantic memory was investigated in eight confabulating and nine non confabulating amnesic subjects, and in 17 matched control subjects, by using a personal and an historical version of the Crovitz [Unconstrained search in long term memory, Paper presented at the meeting of the Psychonomic Society, St Louis, MO, 1973] cue-word test. In response to cue words such as letter or battle subjects had to describe in detail, respectively, a related event from their personal lives or from history before their birth. We found that a subset of amnesic subjects, those with presumed damage or dysfunction in the region of the ventromedial frontal cortex, would confabulate in response to these cues. Their confabulations involved semantic, historical memories, as much as episodic, personal ones: and that distortions of content were at least as common as those of time. Even when not confabulating, they had much more difficulty than other amnesic subjects in recovering memories related to these cues. In confabulating, as compared to non-confabulating, amnesic subjects, prompting led to an increase in confabulations but also to greater recovery of veridical memories. By comparison, non-confabulating amnesic subjects whose memory loss was as severe as that of the confabulators, had a milder deficit on the personal as well as the historical cue-word test. They benefited from prompting somewhat more than matched control subjects. These results suggest that confabulation is associated with impaired strategic retrieval processes resulting from damage in the region of the ventromedial frontal cortex. These strategic retrieval processes help initiate and guide search in episodic and in semantic memory and they help monitor and organize the output from those systems. PMID- 9226663 TI - False recognition after a right frontal lobe infarction: memory for general and specific information. AB - We previously reported a case study of a man with right frontal lobe damage, BG, who showed extraordinarily high false alarm rates on remember-know recognition tests (Schacter, D. L. et al., Neuropsychologia, 1996, Vol. 34, pp. 793-808). Experiment 1 extends his high false alarm rate to yes-no recognition tests. BG typically gives false 'remember' responses on remember-know tests, and this pattern was uninfluenced when he was asked to explain the basis for his 'remember' responses (Experiments 2 and 3). When BG was given a semantic encoding task, he stopped giving 'remember'-based false alarms (Experiment 4). Signal detection analyses revealed that BG had a discrimination deficit and an abnormally liberal response bias (especially for 'remember' responses) in most conditions. Overall, BG's high false alarm rate is interpreted as reflecting an over-reliance on the general similarity between a test item and the study episode. PMID- 9226664 TI - Fragments of memory. PMID- 9226665 TI - Surgical management of brain-stem cavernomas. AB - We present a series of seven patients who were operated on for symptomatic brain stem cavernomas. The following approaches were used: medial suboccipital (N = 4), lateral suboccipital (N = 1), subtemporal-transtentorial (N = 1), and frontal transcortical-transventricular-subchorioidal-trans velum interpositum (N = 1). Intraoperative motor (N = 4) and somatosensory (N = 1) evoked potential monitoring revealed temporary changes in 3 patients. Immediately postoperatively, the following additional deficits were observed in 6 patients: oculomotor nerve paresis (N = 2), abducens nerve paresis (N = 3), facial nerve paresis (N = 2), deafness (N = 1), and increased ataxia (N = 3). One patient died due to septic complications not related to surgery. After a mean observation time of 2 years, 2 patients had improved, 3 were unchanged, and 1 patient deteriorated as compared to his preoperative status. In conclusion, surgical treatment of brain-stem cavernomas, although carrying a significant risk of temporary neurological deterioration is recommended in symptomatic patients in whom the cavernoma seems to reach the surface of the brain-stem. Intraoperative functional topographic mapping and monitoring have proven useful tools lowering the surgical risks in these patients. PMID- 9226666 TI - Multiple communicating intradural arachnoid cysts: usefulness of myelography and myelo-computed tomography using both lumbar and cervical punctures. Case report. AB - We report a case of multiple communicating intradural cystic lesions. Magnetic resonance imaging did not demonstrate the lesions. Neuroradiological diagnosis of the intradural arachnoid cysts was made from myelography and myelo-computed tomography using both lumbar and cervical punctures. These procedures give us useful information about flow dynamics in the spinal subarachnoid space. PMID- 9226667 TI - Surgical indications and prognosis in spinal metastases. AB - Based on the catamnestic evaluation of our own case material and the compilation of recent series from the literature, the indication for surgical intervention in spinal metastases is considered in the light of factors influencing the prognosis of the underlying malignant disease. PMID- 9226668 TI - Idiopathic normal-pressure hydrocephalus in adults: result of shunting correlated with clinical findings in 18 patients and review of the literature. AB - The authors describe their findings in a study aimed at identifying clinical prognostic factors in treatment of idiopathic normal-pressure hydrocephalus. The study comprised 18 adult patients submitted to surgery for ventriculo-peritoneal shunting. The findings that emerged from this series of patients were compared with those reported for the 381 published cases. In our group of 18 patients, average age was 65 years and the average duration of clinical history was 47 months (median 18 months). Follow-up ranged from 3 to 5 years (median 4.2 years): 12 patients improved (9 completely) and 6 presented stable neurological deficits. The factors that had a statistically significant influence on outcome were a short clinical history (less than 6 months) (p = 0.05) and a clinical onset without dementia (p = 0.03). Patients with medium-grade preoperative ventricular enlargement always made a complete functional recovery after surgery (p = 0.2). PMID- 9226670 TI - Percutaneous radiofrequency rhizotomy of lumbar spinal facets: the results of 46 cases. AB - The results of percutaneous radiofrequency rhizotomy of lumbar spinal facets in 46 patients followed at least three months (mean 15 months) are reported and compared with those reported previously. Satisfactory pain relief three months after the procedure was achieved in 36.4 percent of patients without operations and in 41.7 percent of patients, with operations other than fusion. No patient had previously undergone fusion. Treatment of low-back pain by using radio frequency thermocoagulation of spinal facets is a simple, safe, and well tolerated procedure. It can be used to relief of pain in spite of decreasing rates of success within the follow-up period. PMID- 9226669 TI - Usefulness of two-dimensional time-of-flight MR angiography combined with surface anatomy scanning for convexity lesions. AB - Thirty-eight patients with convexity lesions were studied prospectively with the two-dimensional time-of-flight (2D-TOF) magnetic resonance angiography (MRA) method. Of these 21 cases had additional surface anatomy scanning (SAS) and 7 cases had three-dimensional phase contrast (3D-PC) MRA. The findings were compared during surgery and the predictability of 2D-TOF evaluated. 2D-TOF was obtained with 2 mm slice thickness after the administration of contrast media for routine magnetic resonance imaging (MRI). Cortical veins were visualized with a good resolution with a scan time of only 5 minutes. The tumor was also visible in the background, due to enhancement, and thus the tumor-vessels relation was shown. Slow-flow vessels were also adequately seen. SAS was done at the same sitting with fast spin echo (FSE) with a scan time of 3 minutes. Once both images were incorporated, information on gyri and their relation to the lesions and vasculature could be obtained from a single image. We found 2D-TOF alone, or at times in combination with SAS, useful for planning of operation for convexity lesions. PMID- 9226671 TI - Multiple meningiomas of the central nervous system without the stigmata of neurofibromatosis. Clinical and therapeutic study. AB - Multiple meningiomas are relatively rare tumors without known neurofibromatosis. In this paper, such eight cases of multiple meningiomas as described by CUSHING and EISENHARDT are presented. Certain aspects of diagnosis and surgical management of this rare condition are discussed with particular emphasis on the importance of the distinction among multiple meningioma, meningiomatosis, or recurrences of these tumors. Fortunately, many of these patients tolerate multiple surgical interventions well, although the removal of these tumors in critical areas is a difficult problem. Thus, we think that it is important to examine and supervise all patients who have had a meningioma for a possible occurrence of a second meningioma. PMID- 9226672 TI - Effects of nerve growth factor on acetylcholinesterase activity of the proximal stump of the transected sciatic nerve. AB - 28 rats were studied. They were divided into four groups of 7 rats of each one day control animals (Group A), seven-day control animals (Group B), one-day nerve growth factor (NGF)-treated animals (Group C) and seven-day NGF-treated animals (Group D). In all animals, the right sciatic nerve was explored and completely transected using microscissors under a microscope. In NGF-treated animals, NGF (0.1 mg/kg NGF) was injected subcutaneously. In control animals, there was a statistically significant decrease in AChE activity in the proximal stump of the transected sciatic nerve at 7-day measurements (p < 0.05). In the NGF-treated groups, mean AChE activity in the proximal stump of the transected sciatic nerve was decreased at the 7-day measurement. AChE activity difference between control and NGF-treated groups, measured on the 7th day, are statistically significant (p < 0.01). These results demonstrate the effect of NGF on the cholinergic system. PMID- 9226673 TI - Surgical removal of cavernous angioma in the medulla oblongata. A case report. AB - Recent advances in neuroradiology have enabled us to approach cavernous angioma in the medulla oblongata, rather rare vascular lesion in the central nervous system. We describe a such surgical case without additional neurological symptoms and discuss surgical indications in this paper. A 61-year-old woman presented with vertigo and swallowing disturbance. T1-weighted magnetic resonance image (MRI) showed a low intensity mass in the dorsolateral portion of the medulla oblongata, and T2-weighted imaging revealed a hemosiderin rim surrounding the lesion. Angiography showed no abnormalities. Surgery using far lateral approach achieved complete removal of the mass and hematoma. Histological examination of the surgical specimen disclosed cavernous angioma. This case suggests that direct surgery can be recommended for cavernous angioma located in the dorsal or lateral medulla oblongata to remove the hematoma and angioma if bleeding clearly provokes neurological symptoms. PMID- 9226674 TI - Primary Ewing's sarcoma of the temporal bone with intracranial, extracranial and intraorbital extension. Case report. AB - Primary cranial Ewing's sarcoma is a very rare malignant tumor. Primary temporal bone Ewing's sarcoma with intraorbital extension is also a very rare condition. In this article we describe a case of primary Ewing's sarcoma of the temporal bone which extended into intracranial, extracranial and intraorbital compartments; manifesting with a swelling in the zygomatic fossa and proptosis of the right eye. The tumor was excised partially, and the patient underwent radiochemotherapy. The patient died 6 months after the therapy, with lung metastasis. The case is discussed with the pertinent literature. PMID- 9226675 TI - Intracerebral Aspergillus abscess: case report and review of the literature. AB - Intracranial aspergillosis is a rare pathologic condition, difficult to treat and often fatal, which generally affects immunodepressed patients. A case of brain abscess secondary to pulmonary localization in a patient with a non-Hodgkin lymphoma is described. The most significant clinico-pathological findings of intracranial aspergillosis are examined in the light of the relevant literature. PMID- 9226676 TI - Intraparenchymal schwannomas of the central nervous system: an additional case report and review. AB - Intraparenchymal location of schwannomas in the central nervous system (CNS) is rare. Occasional cases involving the cerebrum, cerebellum, brain stem, and spinal cord have been reported. We report here an additional case of thoracic intramedullary schwannoma in a 42 year old woman. The literature concerning intraparenchymal schwannomas in the CNS is reviewed and discussed. PMID- 9226677 TI - Dissecting aneurysms of the anterior cerebral artery and accessory middle cerebral artery. Case report. AB - A 66-year-old woman presented with dissecting aneurysms of the anterior cerebral artery (ACA) and accessory middle cerebral artery (MCA) manifesting as subarachnoid hemorrhage but without radiological evidence of the dissecting aneurysms. Intraoperative observation revealed that the vessel walls were dark purple in color, a typical finding of dissecting aneurysm. The abnormal A1 segment was trapped and the dissecting aneurysm of the accessory MCA was wrapped. In the case of SAH of unknown origin, dissecting aneurysm should always be kept in mind even if the angiogram does not show any abnormal finding. This is the first reported case of dissecting aneurysm of the accessory MCA. PMID- 9226678 TI - In vivo and in vitro cellular response to Schistosoma japonicum eggs in hosts with differing susceptibilities. AB - A comparative study of the cellular response to Schistosoma japonicum eggs was conducted in order to explore its significance, using hosts with differing susceptibilities to the parasite. In experimentally induced, synchronized hepatic granuloma formation, animal species formed each characteristic feature of the granulomas, and the magnitude of tissue reaction was significantly larger in highly susceptible hosts, such as mice and hamsters, while less susceptible hosts, such as rats and quails, formed smaller granulomas. Confluent neutrophils were seen within the tissue lesions for mice and hamsters, while rats and quails showed obviously scanty neutrophils. Guinea pigs failed to develop any granulomas. When splenic cells and bone marrow cells were used for in vitro granuloma formation, bone marrow cells showed markedly higher reactions than splenic cells from naive or sensitized animals and the reactivities of bone marrow cells from susceptible hosts, mice and hamsters, were clearly higher than those from rats, indicating similar results to those of in vivo granuloma formation. This study indicates that the in vivo and in vitro cellular response to S. japonicum eggs varies greatly according to the host's susceptibility, independent of whether the host is a naive or sensitized animal. Our results seem to support the concept that the parasites exploit the host immune system in order to complete their life-span. PMID- 9226679 TI - Function and expression of a human idiotypic network in Schistosomiasis japonica. AB - The cross-reactive idiotype (Hu-SJ-CRIM) is defined by polyclonal human anti idiotypic antibodies derived from chronically S. japonicum infected patients. The present study shows that serum levels of Hu-SJ-CRIM expressed by antibodies to S. japonicum soluble egg antigen (SEA) are associated with acute infection and hepatosplenic disease. Xenogeneic anti-idiotypic antisera (anti-Hu-SJ-CRIM) suppressed human lymphocyte blastogenesis to SEA in vitro by 47-82% (P < 0.05). These anti-idiotypic antibodies also suppressed in vitro granuloma formation induced by SEA coated heads in a dose dependent manner. This immunosuppression was antigen specific in that mitogen (PHA) or non-related antigen (PPD) induced blastogenic responses were not suppressed. Surprisingly, anti-idiotypic antibodies (anti-SJ-CRIM), which describe the mouse correlate CRIM were not suppressive in the human blastogenesis or in vitro granuloma formation assays. These data indicate a dichotomy in the function and specificity of the idiotype/anti-idiotype human and murine immune networks in S. japonicum infection. Thus, only the patient derived molecules and serology form the basis for an immunoregulatory network in Schistosomiasis japonica. PMID- 9226680 TI - Genetic control of immunity to Heligmosomoides polygyrus: presentation of promiscuous antigens to parasite-specific T cell hybridomas. AB - The role of MHC class II in the presentation of Heligmosomoides polygyrus antigens has been investigated, using a number of T cell hybridomas produced in A and E positive and negative mice. By using fixed and irradiated antigen presenting cells (APC), further evidence has emerged, to support earlier data, that there can be differential processing requirements during the presentation of H. polygyrus antigens by A and E molecules. In concordance with these earlier observations, this work provides further evidence than individual T cells can respond to antigen when presented by more than one MHC molecule. Previously, this evidence has been restricted to individual MHC molecules of the same haplotype, but these data show that H. polygyrus produces antigens which can be presented by both syngeneic and allogeneic MHC molecules. These antigens do not appear to be synonymous with the previously described H. polygyrus superantigen, as presentation is restricted to specific MHC haplotypes. It is proposed that H. polygyrus may produce these antigenic molecules as part of its strategy to manipulate the host immune system. PMID- 9226681 TI - Vaccination against hydatidosis using a defined recombinant antigen. AB - Echinococcus granulosus is the causative agent of hydatid disease in humans and animals. Natural transmission of the parasite occurs between dogs as definitive hosts and animal intermediate hosts. There is an urgent need for improved methods to control the parasite's transmission. Here we describe the development of a vaccine based on a cloned recombinant antigen from the parasite egg (oncosphere). Sheep-vaccinated with the antigen, designated EG95, are protected (mean 96-98%) against hydatidosis developing from an experimental challenge infection with E. granulosus eggs. The vaccine will provide a valuable new tool to aid in control of transmission of this important human pathogen. It also has the potential to prevent hydatid disease directly through vaccination of humans. PMID- 9226682 TI - Characterization of T-cell immune responses of Echinococcus multilocularis infected C57BL/6J mice. AB - Specific and non-specific parasite-induced changes in lymphocyte responses were analysed in C57/BL/6J mice after intrahepatic infection with Echinococcus multilocularis. Spleen cells harvested at selected times after infection were in vitro stimulated with mitogens or a crude soluble parasite extract (EmAg) at an optimized dose. Cell proliferative responses to Con-A were not modified by the infection over the first 22 weeks. In contrast, LPS-induced responses were decreased from the 13th week. A strong CD4+ proliferative T-cell response to the parasitic extract of infected mouse spleen cells was observed at the early stage of infection. This response then progressively decreased but remained significantly higher than that of control mice until the 19th week of infection. Cytokine production was investigated after in vitro EmAg stimulation of spleen cells. IFN-gamma, IL-2, IL-5 were produced within the first weeks after infection whereas the detection of IL-10 was slightly delayed. Thus, the promotion of the disease does not appear associated with the expansion of one rather than another T-cell subset in C57BL/6J mice. A general immunosuppression affecting both mitogenic and parasite-specific T-cell responses was observed at the end of the infection. PMID- 9226683 TI - Glutathione-binding proteins of Setaria digitata: antibody responses in human infected with Wuchereria bancrofti. AB - Glutathione-s-transferase activity was determined in filarial parasites. The activity was detected in adult stages of cattle parasite Setaria digitata. It was absent in other stages of Setaria and also in infective larval stages of Wuchereria bancrofti and Brugia malayi. The activity was enhanced about twenty five fold following purification of adult setaria extracts on glutathione agarose column. Antibody (IgG and IgM) levels to the affinity purified proteins (SdGBP) were detected predominantly (90%) in Wuchereria bancrofti infected individuals compared with normal residents of endemic regions. IgA and IgE responses could not be detected. Filarial sera in contrast to non-filarial caused reduction in the enzymatic activities of Sd GBP. Microfilaraemic sera after diethylcarbamazine treatment resulted in enhanced reduction of enzymatic activity. PMID- 9226684 TI - Microfilarial clearance in loiasis involves elevation of Th1 and Th2 products and emergence of a specific pattern of T-cell populations. AB - Diethylcarbamazine (DEC) induced clearance of microfilaraemia in loiasis is associated with severe posttreatment reactions. To define the switch from hypo- to hyper-responsiveness associated with DEC treatment, phenotypic alterations of T-lymphocytes, characterized by flow cytometry, and cytokines, determined by enzyme linked immunosorbent assay, were monitored in a microfilaraemic patient. In contrast to reports on onchocerciasis and lymphatic filariases, no elevation of interleukin (IL)-6 and tumour necrosis factor (TNF)-alpha was observed. The most severe side effects coincided with an elevation of interferon (IFN)-gamma on day 3, followed by IL-10, transforming growth factor (TGF)-beta 2 and macrophage inflammatory protein-1 alpha (MIP-1 alpha) peaking on day 5. Phenotypically, T cell activation markers CD38, CD54 and CD25 were significantly expressed before treatment, with high CD38 expression still existing one year after clearance of microfilaraemia. Treatment-related increases were observed with anti-CD122, anti HLA-DR and anti-CD69. CD28 was expressed before treatment on almost 100% of CD4+ and CD8+ T cells and dropped to 20% by day 5, reaching again baseline levels on day 21. Furthermore, there emerged 20% TCR alpha beta+/CD3+ T cells and 10% anti beta V5(c)+ T cells, altogether indicating a specific pattern of T-helper (Th) 1 and Th2 cytokines as well as expansion of certain pauciclonal T-cell populations in response to microfilarial clearance. PMID- 9226685 TI - Malaria cellular immune responses in neonates from Cameroon. AB - T cell responses to leucoagglutinin, PPD, and seven Plasmodium falciparum blood stages antigens were investigated in 164 cord blood samples from Cameroonian neonates. In vitro T cell responses were measured by lymphocyte proliferation, and IL-2, IFN-gamma, and IL-4 release in the presence of crude schizont extract, purified Pf155/RESA protein, and synthetic peptides from Pf155/ RESA. Following culture in presence of leucoagglutinin or PPD, proliferation and cytokine production were very low, as compared to adults from the same area. Interestingly, following stimulation of cord blood lymphocytes by malaria antigens, the percentage of responders and the mean level of positive responses were of the same order than those observed in adults for IL-2 production, while proliferative and IL-4 responses were only marginally decreased. Conversely, IFN gamma production was highly reduced, as compared to adults. Our results demonstrate that prenatal immune priming to malarial antigens is common in this area and that the fetal immune system is able to respond to antigenic stimuli, as cells proliferate and generate cytokines. As cord blood lymphocytes may be induced to differentiate into effector cells producing predominantly Th1 or Th2 cytokines, malaria during pregnancy might direct the functional capacity of fetal T cells to respond to further infection. PMID- 9226686 TI - Increased frequency of HLA-DR3 and complotype SCO1 in Mexican mestizo children with amoebic abscess of the liver. AB - The increase of HLA-DR3 and complotype SCO1 previously found in Mexican mestizo adults with E. histolytica amoebic abscess of the liver, was also found in Mexican mestizo children of either sex with the same disease, when compared to the healthy control population (adults and/or children) of the same ethnic and socioeconomic background. This HLA and complotype pattern was not found in Mexican Mestizo patients with amoebic rectocolitis. No linkage disequilibrium was found between these and the other MHC determinants tested in this survey. Thus, HLA-DR3 and SCO1 may constitute primary, independent risk factors, not for any kind of amoebic tissue invasion (i.e. amoebic rectocolitis), but specifically for amoebic liver abscess, irrespective of age or sex. The possibility of linkage disequilibrium with other factors (i.e. the TNF family) within or close to the MHC that were not tested in this study, is discussed. Children with amoebic liver abscess revealed a significant increase in HLA-DR5, and the absence of HLA-DR6 when compared to adults with amoebic liver abscess, suggesting that at least in this ethnic group these class II HLA traits may contribute to some of the peculiarities of pediatric amoebic liver abscess as opposed to the adult version of this disease. HLA-DR3, SCO1, but also HLA-DR5 and HLA-DR6 have all been associated with certain forms of immune-dysfunction, and may thus contribute to some of the clinical and immunological features of this parasitic disease. PMID- 9226687 TI - Correlation of antigen specific IgG and IgG(T) responses with Anoplocephala perfoliata infection intensity in the horse. AB - There is increasing interest in the application of serological methods to macro parasite infections to indicate infection intensity, which in turn is related to pathogenicity. Colic is the single most important cause of mortality in horses and there is evidence that a proportion of colic cases are associated with infection with the intestinal cestode Anoplocephala perfoliata. In order to develop better tools to investigate this association, the correlation between antigen-specific equine IgG and IgG(T) and infection intensity of A. perfoliata was investigated. Affinity purification of a 12/13 kDa protein doublet from crude excretory/secretory (E/S) products, and its use in enzyme linked immunosorbent assays (ELISA) is described. Its use in the immunodiagnosis of equine cestodosis and the correlation of anti-12/13 kDa IgG and IgG(T) with parasite burden is investigated using sera from 94 horses of known tapeworm infection intensity. The anti-12/ 13 kDa IgG and IgG(T) ELISAs gave correlation coefficients with infection intensity of 0.56 and 0.63 respectively. Linear regression analysis also indicated that anti-12/ 13 kDa IgG(T) was the best predictor of infection intensity. The decay of anti-12/13 kDa IgG(T) in horses following the elimination of A. perfoliata is demonstrated for four horses. Specificity of the anti-12/13 kDa IgG(T) ELISA is investigated with sera from 33 A. perfoliata negative horses with other helminth infections. Immunoblotting studies demonstrate no cross reactivity between A. perfoliata 12/13 kDa antigen and the protein antigens of other helminths. It is concluded that assay of anti-12/13 kDa IgG(T) provides a useful tool for the assessment of A perfoliata infection intensity for clinical diagnosis and for epidemiological studies. PMID- 9226688 TI - Host-protective fragments and antibody binding epitopes of the Taenia ovis 45W recombinant antigen. AB - The protective efficacy of a recombinant Taenia ovis vaccine antigen, 45W, was compared in sheep vaccine trials with antigen expressed by the full length 45W cDNA and by incomplete copies of the cDNA. Vaccine, trials were also carried out using antigen expressed by a cDNA (45S) having a sequence similar, but not identical, to 45W. Stability of the 45W antigen expressed in Escherichia coli was found to be increased after deletion of cDNA sequence encoding 19 COOH-terminal amino acids. This truncated form of the antigen was designated 45WB/X. Vaccination of sheep with antigen expressed by 45W, 45WB/X, as well as full length 45W and 45S cDNAs, induced high levels of protection. Vaccination with antigen expressed by an incomplete copy of the 45S cDNA clone did not induce protection. Comparison of deduced amino acid sequences for these clones suggests that the host-protective epitope(s) of the 45W antigen occur on either or both of the 23 and 9 amino acid peptides at the amino and carboxyl termini of 45W, respectively. Antibody binding epitopes of 45W were investigated in ELISA using overlapping 9 amino acid peptides. Protection was found to correlate with the induction of antibody to two 9 amino acid peptides. PMID- 9226689 TI - Identification of Plasmodium falciparum MSP-1 peptides able to bind to human red blood cells. AB - To determine amino acid sequences of the Plasmodium falciparum MSP-1 protein that interact with red blood cell membranes in a specific receptor-ligand interaction, 78 sequential peptides, 20 amino acids long and spanning the entire length of the molecule, were synthesized and analysed with a specific binding assay developed for this purpose. Results show that peptides based on conserved and dimorphic regions of MSP-1, interact with human red blood cells (RBCs). This interaction occurs predominantly with peptides contained within the MSP-1 proteolytic fragments of 83 kDa, 38 kDa, 33 kDa and 19 kDa. Affinity constants of these peptides were between 140 and 250 nM. Peptide-RBC binding post enzyme treatment showed that the RBC receptors are not sialic acid dependent and appear to be proteic in nature. Some of these peptides inhibited merozoite invasion of RBCs yet did not inhibit intraerthrocytic development. These peptides, in conjunction with those from other merozoite surface proteins, may be used to rationally design a second generation of synthetic peptide-based malaria vaccines. PMID- 9226690 TI - Humoral immune responses in Gambians to Pfs16, an immunodominant, Plasmodium falciparum integral membrane protein. AB - Naturally acquired humoral immune responses to Pfs16, an integral membrane protein expressed in Plasmodium falciparum gametocytes and sporozoites, were investigated in The Gambia. A high prevalence of antibodies to this molecule was detected by peptide ELISA. Ninety-three per cent of the people taking part in a survey at the end of the rainy season (November) had serum antibodies to one or more synthetic peptides spanning the sequence: 88% reacted with one particular peptide sequence (IMLIILSGIVGFKVK) whereas only one out of ten non-Gambians (taking anti-malarial prophylaxis with no history of infection) reacted with the peptide. Epitope mapping with mouse MoAbs has shown that this peptide is located on a part of the molecule differing from immunodominant regions of the molecule identified in a previous study in Papua New Guinea. PMID- 9226691 TI - Nitric oxide synthase activity in malaria-infected mice. AB - Nitric oxide (NO) is implicated in a variety of major cellular functions including defence from invasion by microbical pathogens. Evidence has been presented suggesting that it is an important mediator of protection in the early non-specific responses to malaria in mice infected with Plasmodium chabaudi (Taylor-Robinson et al. 1993). Other data from in vitro studies on the asexual stages of human parasite Plasmodium falciparum indicated that while nitric oxide itself may not be inhibitory to parasite development, its downstream products do have some anti-plasmodial activity (Rockett et al. 1991) and these could be generated by macrophages (Gyan et al. 1994). Similarly, the sexual phases of both rodent (Motard et al. 1993) and human malaria (Naotunne et al. 1993) are reportedly susceptible to the toxic effects mediated by nitric oxide generated by blood leucocytes in the course of transmission to the mosquito vector. PMID- 9226692 TI - Presence of antibodies in the aqueous humour and cerebrospinal fluid during Leishmania infections in dogs. Pathological features at the central nervous system. AB - In the present paper we show that in dogs, naturally infected with Leishmania infantum, the aqueous humour and the cerebrospinal fluid contain anti-Leishmania IgGs and that the specificity of antigen recognition of these fluids is similar to that of the sera. We also show that in the encephalon and cerebellum of these dogs there is a pathological sponge-like reaction accompanied by neuronal degeneration, mobilization of glial cells together with accumulation of amyloid deposits. The interstitial and intravascular deposition of IgGs and Leishmania antigens in choroid plexus suggest that in these animals there is a failure of the blood-cerebrospinal and ciliary bodies filtration barriers which may allow the transfer of anti-Leishmania IgGs from the blood stream to these fluids. We suggest that the failure of the blood-cerebrospinal barrier and the in situ concentration of anti-Leishmania IgGs and antigens in brain tissues may predispose to the pathological features detected in this compartment. PMID- 9226694 TI - Carbohydrates on the surface of Echinococcus granulosus protoscoleces are immunodominant in mice. AB - A carbohydrate enriched soluble fraction (CHP) was prepared by mild treatment of viable Echinococcus granulosus protoscoleces (PSC) with the enzyme endoglycosidase-F (endo-F) and characterized by SDS-PAGE, glycoside- inhibition ELISA, and immunoblotting. Three groups of four BALB/c mice were immunized with the CHP, with the remaining deglycosylated PSC (DGP) and with dead PSC (DPSC) to analyse the relative immunogenicity of carbohydrates on the surface of PSC. A fourth sentinel group was not immunized. The antibody response was analyzed during primary and hyperimmune responses. Specific antibody titres (IgG and IgM) against somatic PSC antigens (PSA) were evaluated by ELISA and their antigen recognition pattern by immunoblotting, discriminating carbohydrate and peptide specific antibody responses by periodate treatment of PSA. The avidity index of those antibodies and the titer of non-specific immunoglobulins during the whole protocol were evaluated by ELISA in vitro mitogenic activity of CHP was also evaluated. The results indicated: 1) immunodominance of surface carbohydrate epitopes from PSC, 2) predominant IgM and low avidity response against these epitopes, 3) a dramatic increase of non-specific antibody titres and 4) in vitro mitogenic activity of CHP. PMID- 9226693 TI - Influence of adjuvants on murine immune responses against the C-terminal 19 kDa fragment of Plasmodium vivax merozoite surface protein-1 (MSP-1). AB - The immunogenicity of a yeast-expressed 19 kDa fragment of P vivax MSP-1 in the presence of different adjuvant formulations was evaluated. ICR mice were immunized with the 19 kDa antigen, using Freund's, alum, and block copolymer P1005 in water-in-oil (W/O) or oil-in-water (O/W) emulsions with or without detoxified lipopolysaccharide (RaLPS) as adjuvants. Five weeks following immunization with the antigen, mice were boosted with asexual blood-stage antigens. Three weeks after the last immunization with the 19 kDa antigen, mice from the Freund's group and most groups that received P1005 as adjuvant had higher total IgG titres than those that received alum as adjuvant or antigen alone. Antibody responses after the antigen immunization were predominantly of the IgG1 isotype, but mice in the Freund's and P1005 (W/O or O/W emulsion with or without RaLPS) groups also had high titres of IgG2a and IgG2b. Antibody titres against merozoites increased in all groups after the parasite antigen boost. IgG2a levels in the group that received antigen in P1005 plus RaLPS in the W/O emulsion were higher than those receiving Freund's, alum or the other copolymer adjuvants. The high IgG2a titres in this group were associated with reduced IL-10 production. PMID- 9226695 TI - Immunological responses to antigen B from Echinococcus granulosus cyst fluid in hydatid patients. AB - The immunological reactivity of Echinococcus granulosus antigen B was evaluated in 30 hydatid patients. Antigen B was purified from sheep hydatid cyst fluid by electroelution from a non-reducing SDS-PAGE gel (AgB). In ELISA and immunoblotting (IB), determining antibody production in sera from patients with hydatid disease and with other parasitic infections, purified AgB showed higher specificity than a partially purified antigen named pH5PPT (100% vs 83% in pH5PPT ELISA and 58% in pH5PPT-IB). AgB-IB achieved higher sensitivity than AgB-ELISA (80% vs 63%). All AgB-IB positive sera recognized the 12 kDa subunit. Qualitative AgB-IB assessment of IgG isotype responses identified IgG4 as the predominant isotype (87%). The other isotypes showed a lower percentage of positive reactions: IgG1, 33%; IgG2, 21%; and IgG3, 17%. PBMC proliferative assay revealed a cellular response to AgB in 100% of patients' PBMC. These findings confirm antigen B, especially its smallest subunit, as a good diagnostic molecule. PMID- 9226696 TI - Influence of parasite presence on the immunologic profile of peripheral blood mononuclear cells from chagasic patients after specific drug therapy. AB - The aim of this study is to evaluate the effects of parasite clearance on the immunological profile of peripheral blood mononuclear cells (PBMC) from chagasic patients submitted to specific drug therapy. PBMC were examined by flow cytometry and proliferative responsiveness to Trypanosoma cruzi-related stimuli. Three groups of patients were studied: not treated (NT), treated not cured (TNC) and cured (C). All data were compared to values from uninfected individuals (NI). NT displayed a lower percentage of CD3+ cells as compared to NI, while TNC and C had mean values that were between those from NI and NT. Infected patients had double the percent of CD3+ HLA-DR+ cells, independent of the efficacy of the treatment. Thus, absence of circulating parasites did not reduce T cell activation in Chagas' disease. NT displayed a higher percentage of CD5+ B cells as compared to NI, while TNC and C had mean values between those from NI and NT. In contrast to the phenotypic data, the in vitro mean proliferative responses to parasite related stimuli of PBMC from C were reduced to the low mean levels observed in NI. These striking differences were statistically different from the high responses seen in NT and TNC. Our data suggest that proliferative responses of PBMC from C reflect immunological changes due elimination of parasite. However, successful treatment did not alter the levels of peripheral T cell activation. PMID- 9226697 TI - Two different 8 kDa monomers are involved in the oligomeric organization of the native Echinococcus granulosus antigen B. AB - The present work describes the purification and characterization of antigen B (AgB), the thermostable lipoprotein from E. granulosus. Native AgB was purified to homogeneity by a new strategy involving adsorption on DEAE-Sepharose, followed by immunopurification. The purified antigen was analysed using mapped monoclonal antibodies (MoAbs) and peptide isolation by in situ digestion in gels after SDS PAGE. Epitope mapping of 7 MoAbs using PEPSCAN, synthetic peptides and competition studies, revealed that six of them defined epitopes which clustered the N-terminal extension of a 8 kDa subunit of AgB, whilst the remaining one reacted against the stretch RGLIAEGE, corresponding to the C-terminus. The epitopes defined by the seven MoAbs were found to be present in all the subunits. Furthermore, the similarities of the peptide finger prints obtained by HPLC analysis and amino acid sequencing of tryptic peptides isolated from the 8, 16 and 24 kDa subunits, indicated that they have most if not all the amino acid sequence in common. We also found evidence that the band representing a component of an apparent molecular weight of 8 kDa in SDS-PAGE, believed to be the smallest subunit of AgB, contained at least two components, which may constitute the building blocks of the higher molecular weight subunits. PMID- 9226698 TI - Purification and N-terminal amino acid sequencing of Echinococcus granulosus antigen 5. AB - Antigen 5, a major parasite-derived lipoprotein of unilocular hydatid (Echinococcus granulosus) cyst fluid, comprises subunits of 56-70 kDa which dissociate on disulphide bound reduction to two subunits of approximately 25 and 38 kDa on SDS-PAGE. The 38 kDa component is recognized as a potentially important diagnostic marker for cystic hydatid disease. Single dimensional SDS-PAGE, two dimensional (2-D) gel electrophoresis, modified '2-D' gel electrophoresis, immunoaffinity-purification and HLPC were employed to purify antigen 5. Subsequent N-terminal sequencing suggested that antigen 5 isoforms exist due to the identification of a single amino acid sequence with alternative residues at some positions. An 18 amino acid consensus N-terminal sequence was derived for antigen 5 as a result of comparing the sequences obtained by the different fractionation methods. No homology was revealed to any previously identified protein including putative antigen 5 amino acid sequences. A synthetic peptide, mimicking the N-terminal consensus sequence, did not bind with patient sera (confirmed positive to antigen 5) or an anti-antigen 5 MoAb. Protein sequences (16 residues compared) for the 38 kDa subunit from sheep and horse (representing different strains of E. granulosus) cyst fluids were very similar except for differences at three positions. 2-D and modified '2-D' gel electrophoresis, in combination with immunoblot analysis, indicated the presence of more than one protein in the 38 kDa region of SCF capable of binding the anti-antigen 5 MoAb. PMID- 9226699 TI - The flagellar fraction of Trypanosoma cruzi depleted of an immunosuppressive antigen enhances protection to infection and elicits spontaneous T cell responses. AB - The flagellar fraction (FF) of Trypanosoma cruzi can be separated by immunoaffinity chromatography in two fractions with balanced but opposite immunological effects. The immunoaffinity purified fraction has immunosuppressive activity mediated at least partially by TGF-beta (Hansen et al., submitted). Here we report that the fraction depleted of immunosuppresive antigens (FT) administered with iscom-matrix as adjuvant provides enhanced protection to an infection challenge in immunized mice. In vitro, the FT but not the FF stimulated resident peritoneal cells to produce IL-1 and IL-6. In immunized mice, the FT elicited higher levels of antigen-specific IgG2a than the FF as well as broader recognition of T. cruzi antigens. Splenocytes from mice immunized with FT proliferated spontaneously in vitro and secreted TH1 and TH2 cytokines. The protection provided by FT correlates with its capacity to enhance the secretion of IFN-gamma. We postulate that immunosuppressive antigens present in the FF prevent the development of memory cells secreting IFN-gamma through a TGF-beta dependent mechanism. PMID- 9226700 TI - Immunopathology of the uveitis in canine leishmaniasis. AB - Particular immunopathological features and their effects on the vascular permeability of different ocular structures were analysed in two dogs naturally infected by Leishmania infantum. The existence of specific anti-Leishmania immunoglobulin G (IgG) in the aqueous humour was confirmed by the ELISA technique. There was no correlation between antibody levels in the aqueous humour and the related serum. The histopathological study of the eyes showed the existence of lesions in various ocular structures. The ciliary processes, ciliary body, sclerocorneal limbus, iris and lacrimal duct showed intense inflammatory zones with lymphocyte infiltrates, plasmatic cells and macrophages with amastigote forms of Leishmania. In addition vasculitis with dilation and thrombi were also detected in both cases, with consequent oedema and hyalinization. The immunohistochemistry analysis revealed the presence of granular and diffuse IgG deposits in the ciliary body, ciliary processes, sclerocorneal limbus and iris. Furthermore, numerous thrombosed vessels were observed in the sclerocorneal zone and iris. Complement 3 (C3) fraction deposits were not present in the ocular structures. The present data suggest that the ocular lesions may have an immunopathological origin. PMID- 9226701 TI - HPLC fractionated soluble egg antigen from Schistosoma mansoni elicits a heterogeneous human immune response. AB - Peripheral blood mononuclear cells (PBMC) from five chronic schistosomiasis patients, three former patients, a SEA sensitized individual, and normal controls were tested in lymphoblastogenesis assays for their ability to proliferate in response to soluble egg antigen (SEA) and soluble worm antigen preparation (SWAP) from Schistosoma mansoni. Cells from all patients and the SEA sensitized individual gave significantly higher responses than the normal controls when stimulated with SEA and SWAP. However, the chronic patients' SEA responses were much lower than those of the former patients and the SEA sensitized individual. When cells from the same donors were tested in the in vitro granuloma assay, all produced significant granulomatous responses except the normal controls. Once it was established that all individuals in the study gave significant lymphoproliferative responses and granulomatous reactions, SEA was subjected to HPLC fractionation to identify immunogenic protein components of SEA. HPLC separation yielded 25 major fractions. SEA responses from the sensitized individual and former patients exhibited broad, unregulated responsiveness including fractions with neutral, less charged proteins while the chronic patients demonstrated a more restricted range of responsiveness. SEA-HPLC fractions 14, 21, and 22 contain the most immunodominant proteins based on cellular proliferation data from reactive individuals tested. PMID- 9226702 TI - Use of blood products in cardiac surgery. AB - The quantity of blood products used perioperatively during cardiac surgery is known to vary widely between institutions. This study looked at the amount of blood products used perioperatively in 74 consecutive elective cardiac operations in one institution. The results are compared with those from other European centres and a cost analysis carried out. On average 2.33 +/- 0.74 (95% confidence interval 1.93-2.77) units of red cell concentrate were transfused perioperatively per patient. Six (8%) patients received no blood products. In addition a number of preoperative factors were studied in an attempt to identify predictors of transfusion requirements. Age, preoperative haemoglobin, female sex and red cell mass were all found to have some predictive value. In the face of increasing demands on a limited supply of blood products we question the need for cross matching more than four units of red cell concentrate in elective cardiac surgery. PMID- 9226703 TI - Neurone-specific enolase and Sangtec 100 assays during cardiac surgery: Part I- The effects of heparin, protamine and propofol. AB - Neurone-specific enolase (NSE) and Sangtec 100 (S-100) (Sangtec Medical, Sweden) assays are designed for clotted samples, but when studying cerebral damage following cardiac surgery, perioperative samples will contain heparin and/or protamine. The lipid emulsion propofol is also frequently used during cardiac surgery and could affect the assays. We, therefore, studied the effects of heparin, protamine and propofol on the accuracy of NSE and S-100 assays in five healthy patients. Blood samples were taken and divided into four groups: normal saline was added to group A; heparin to group B; heparin followed by protamine to group C; and propofol to group D. NSE and S-100 concentrations were measured for all samples. Neither heparin, protamine nor propofol affected the accuracy of S 100 and NSE assays; therefore, samples can be taken throughout operations involving cardiopulmonary bypass without influencing the results. PMID- 9226704 TI - Neurone-specific enolase and Sangtec 100 assays during cardiac surgery: Part II- Must samples be spun within 30 min? AB - Sangtec 100 (S-100) (Sangtec Medical, Sweden) and neurone-specific enolase (NSE) assays are showing promise in the assessment of cerebral damage following cardiopulmonary bypass (CBP). The manufacturer's instructions state, however, that samples must be spun and frozen within 30 min, which is inconvenient for serial studies. We, therefore, investigated whether strong blood samples at room temperature (RT) or 4 degrees C for up to 48 h affected the measured levels. Blood samples were taken before and after CBP in six patients and stored for 15 min, 4, 8, 24 or 48 h at RT or 4 degrees C. S-100 and NSE levels did not alter in either 'before surgery' or CPB samples when stored for up to 48 h at 4 degrees C. There was a small, nonsignificant rise when stored at RT. Samples may, therefore, be collected throughout long operations or stored overnight without affecting NSE or S-100 plasma levels. PMID- 9226705 TI - Neurone-specific enolase and Sangtec 100 assays during cardiac surgery: Part III- Dose haemolysis affect their accuracy? AB - Neurone-specific enolase (NSE) and Sangtec 100 (S-100) are useful for detecting cerebral damage during cardiopulmonary bypass (CPB). However, red cells contain NSE, and the haemolysis frequently caused by CPB could produce a false rise in NSE; S-100 is not found in red cells and should not be affected. We, therefore, compared the effects of haemolysis on NSE and S-100 to see if correction was necessary and possible. From seven patients, serial dilutions of haemolysed red cells were added to plasma (1/64-1/2048), measured for absorption at 540 nm and assayed for NSE and S-100. S-100 concentrations showed no change with haemolysis. Measured NSE increased significantly with haemolysis > 1/512 (an increase of 6.6 micrograms/ml): a correction formula is presented. In 39/48 patients after CPB, mean haemolysis was < 1/256 and would not need any correction. NSE and S-100 assay can, therefore, be used throughout CPB, which allows both glial and neuronal damage to be studied. PMID- 9226706 TI - Preoperative versus postoperative extracorporeal life support in neonatal cardiac patients. AB - The aim of this study is to document our experience with the use of extracorporeal life support (ECLS) in the neonatal cardiac patient, to detect differences in the morbidity and mortality between patients who required ECLS preoperatively and those who required ECLS postoperatively, and to determine the long-term effects of these morbidities. A chart review was undertaken of all neonatal cardiac patients who required ECLS between May 1985 and July 1994 at Kosair Children's Hospital, Louisville, Kentucky. Twenty-three neonatal cardiac patients had received preoperative or postoperative ECLS with an overall survival rate of 35%. Our preoperative and postoperative patients had similar demographics, diagnoses, decannulation rates and survival rates. However, patients receiving postoperative ECLS more frequently required more than two inotropes (p < 0.001), had an increased incidence of renal failure (p < 0.02), had more central nervous system abnormalities on brain imaging studies (p < 0.004), and had a longer hospital stay (p < 0.05). Follow-up testing of survivors yielded normal Bayley Scale of Infant Development (BSID) scores in half of the patients. Survival in the two groups was similar, but a significant difference in morbidity was found. Except for severe intracranial abnormalities, the morbidity was shown to be reversible on follow-up examination. We recommend the continued use of ECLS for neonatal cardiac patients who require preoperative or postoperative support even when severe renal failure ensues or minor abnormalities are detected on brain imaging studies. PMID- 9226708 TI - Extracorporeal circuit design considerations for giant intracranial aneurysm repair. AB - Clinical perfusionists must be able to modify the existing extracorporeal circuit in order to accommodate a specific surgical pathology. The clipping of a giant intracranial middle cerebral artery aneurysm, unapproachable with conventional neurosurgical techniques, required the use of a modified closed cardiopulmonary bypass circuit combined with deep hypothermia and total circulatory arrest. In hospital discussions with anaesthesia, cardiac surgery, neurosurgery, and cardiology enabled an informed team approach directed towards the successful treatment of this complex neurosurgical lesion. PMID- 9226707 TI - Conventional haemofiltration during routine coronary bypass surgery. AB - The use of conventional ultrafiltration during cardiopulmonary bypass (CPB) has been well recognized as an efficient modality of therapy to reverse the effects of deliberate haemodilution. Routine use of the haemofilter was prospectively studied on 60 patients undergoing coronary artery bypass surgery. Group A consisted of 30 patients on whom the ultrafiltrator was used and compared to group B who did not receive the ultrafiltration technique. The COBE 1200 ultrafiltration device was used. The results of the study demonstrated that, in group A, the mean total amount of ultrafiltrate collected during bypass was 2510 +/- 804 ml per patient. The mean 24-h postoperative blood loss was 440 +/- 192 ml in group A and 451 +/- 136 ml in group B. The average bank blood transfused was 0.6 +/- 1.3 units per patient in group A and 0.75 +/- 1.5 units per patient in group B. Postoperative weight gain in group A averaged 3.5 +/- 3.45 lb per patient, compared to 4.8 +/- 3.7 lb per patient in group B. Postoperative length of stay averaged 6.4 +/- 1.5 days per patient in group A and 6.4 +/- 2.1 days per patient in group B. Overall patient charges averaged $33,706 +/- 8348 per patient in group A and $33,041 +/- 7674 per patient in group B. It was concluded that routine use of ultrafiltration during routine coronary artery bypass surgery with CPB offers no improvement in the quality of care nor does it decrease the patient's overall charges. PMID- 9226709 TI - Surgical management of catheter tip thrombus: surgical therapy for right atrial thrombus and fungal endocarditis (Candida tropicalis) complicating paediatric sickle-cell disease. AB - The use of indwelling central catheters for long-term administration of hyperalimentation, chemotherapy or other intravenous therapies is increasing. This unusual presentation of a catheter-induced right atrial thrombus was complicated by fungal infection. We present a case of a paediatric sickle-cell patient who underwent surgical removal of a right atrial thrombus secondary to fungal (Candida tropicalis) endocarditis from an indwelling catheter. Successful thrombus removal utilizing cardiopulmonary bypass and subsequent discharge underscores the importance of surgical therapy in treating this important complication. PMID- 9226710 TI - Cardiopulmonary bypass venous cannulation challenges in a paediatric patient with complex congenital heart disease: a case report. AB - When choosing cannulae for cardiac surgery the two most important factors to be considered are the proposed procedure and the patient anatomy. These factors are especially crucial in paediatric patients with congenital heart disease. A 3-year old, 14-kg male presented to the University of Iowa Hospitals and Clinics with dextro-transposition of the great arteries, atrioventricular canal, left pulmonary stenosis, azygous continuation, bilateral superior vena cavae, partial anomalous pulmonary venous return, left aortic arch and status post-right Blalock Taussing shunt. The complex anatomy presented a surgical dilemma. The course of surgical intervention was determined, a variation of the modified Fontan procedure, and the anatomy of the patient was directly viewed. The surgeon concluded that four venous cannulae were required to provide adequate venous return for the cardiopulmonary bypass (CPB) circuit and a bloodless surgical field. The operation was successfully performed under mild hypothermia with no complications. The patient fully recovered with only mild restrictions on his activity level. This case acutely illustrates the importance of anatomical and procedural awareness when choosing cannulae and cannulation sites for CPB in paediatric patients with congenital heart disease. PMID- 9226711 TI - Isolation and N-terminal sequence of multiple forms of granulins in human urine. AB - Three molecular forms of granulins (also known as epithelins) were isolated, for the first time, in human urine. Their N-terminal sequences, which have also been determined, are identical to those of granulins A and B, previously isolated from human leukocytes, and of granulin F, never isolated before but whose primary structure is known on the basis of the cDNA sequence. The urinary molecules, which show a molecular weight of about 6.5 kDa, are most likely produced by a posttranslational proteolytic processing occurring at the level of the kidney, which appears to be the organ richest in granulin precursor mRNA. The molecular events underlying the precursor processing are unknown, even though the involvement of the protease kallikrein, an enzyme thought to be responsible for the processing of several polypeptidic growth factor precursors, could be hypothesized. Granulins, however, do not show antikallikrein activity. The presence in human urine of isoform F, previously not identified from other human sources, seems to support the hypothesis that mature forms of granulins are generated by an organ-specific precursor processing, on the basis of particular physiological requirements, and to suggest also that this isoform may play "in vivo" an important and specific role in the epithelial cells of the human kidney. PMID- 9226712 TI - Phytoene desaturase: heterologous expression in an active state, purification, and biochemical properties. AB - Conditions were developed for heterologous expression in Escherichia coli of the membrane-bound cyanobacterial/plant-type phytoene desaturase (PDS) from Synechococcus in an active form. Decrease of growth temperature for the transformant to 28 degrees C resulted in an increase of proteins in a supernatant fraction obtained after pressure disruption (20 MPa) of cells and centrifugation. This supernatant in which the highest PDS activity was found was used for purification to a homogeneous protein by ammonium sulfate precipitation and DEAE chromatography. The purified PDS was employed to determine substrate specificity and cofactor requirement. Substrates in addition to phytoene were phytofluene and 1,2-epoxy phytoene which were converted to zeta-carotene and the corresponding 1,2-epoxide. The reaction was stimulated by NAD, NADP, and oxygen. The K(m) values determined for phytoene and NADP were 3.5 microM and 14.3 mM, respectively. PMID- 9226713 TI - One-step HPLC purification procedure for porcine brain 90-kDa heat shock protein. AB - The 90-kDa heat shock protein (HSP90) was purified from porcine brain by a novel single-step purification procedure using diethylaminoethyl high-performance liquid chromatography (HPLC). About 4.8 mg of HSP90 was isolated from 25 g wet wt porcine brain tissue. The purified protein possessed a single moiety on one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis followed by silver staining. Western blotting using monoclonal antibody prepared against human HSP90 confirmed its identity as HSP90. These results indicate that small-scale HPLC purification of HSP90 from porcine brain tissue can be readily accomplished, with high yield, using a convenient one-step purification method. The procedure described in this paper represents a significant improvement in current purification methods for the isolation of HSP90 from porcine brain. PMID- 9226714 TI - Purification, characterization, and kinetics of porcine recombinant dihydropyrimidine dehydrogenase. AB - Porcine recombinant dihydropyrimidine dehydrogenase was purified from Escherichia coli cells using cell disruption, ammonium sulfate fractionation, and chromatography on DEAE-cellulose and 2',5'-ADP-Sepharose. The yield was 60% with a specific activity of 14 units/mg protein. On SDS/PAGE the purified dehydrogenase exhibits a single band, indicating that no proteolytic degradation was taking place during purification. In agreement with the native enzyme, all cofactors, FMN, FAD, NADPH, and two iron-sulfur clusters, have been found. EPR spectra of the reduced dehydrogenase obtained at pH 9.5 are characteristic for two [4Fe-4S]1+ cubanes in dipolar interaction. Quantification of the observed signals indicated 0.95 spins per subunit, showing only partially reduced iron sulfur clusters. The kinetic parameters of the porcine recombinant enzyme are very similar to those of the native enzyme. Thus, it can be concluded that the porcine recombinant enzyme behaves like the native dehydrogenase. PMID- 9226716 TI - Expression and characterization of phosphorylated recombinant human beta-casein in Escherichia coli. AB - Specific serine and threonine residues of recombinant human beta-casein produced in Escherichia coli were shown to be phosphorylated in vivo when human casein kinase II was coexpressed in the same plasmid. All of the phosphorylated forms found in the native protein were also detected in the recombinant protein. The phosphorylation of recombinant human beta-casein was confirmed by immunoblots, fast protein liquid chromatography, urea-polyacrylamide gel electrophoresis, SDS polyacrylamide gel electrophoresis, and liquid chromatography-mass spectrometry. The results indicate that the substrate specificity of casein kinase II in vivo was unaffected in its recombinant form. This is the first demonstration of in vivo phosphorylation of specific residues of a multiphosphorylated protein produced in E. coli with a single plasmid. PMID- 9226715 TI - A general purification protocol for E7 proteins from "high- and low-risk" human papillomavirus types expressed in the yeast Schizosaccharomyces pombe. AB - A purification protocol was developed to obtain human papillomavirus (HPV) type 16 E7 protein expressed in the yeast Schizosaccharomyces pombe. Only three chromatographic steps were necessary to purify the unfused HPV 16 E7 protein to homogeneity (95-99%) as shown by silver staining after polyacrylamide gel electrophoresis. Approximately 0.8 mg of highly purified E7 was obtained from 5 x 10(10) yeast cells. The purified HPV 16 E7 phosphoprotein (Ser 31/32) was refolded and assayed for functionality. Binding to the proteins Rb1 and p107 in vitro and induction of DNA synthesis after microinjection into serum-deprived NIH 3T3 cells suggest that the E7 protein retains some of its biological activities. Most importantly, the purification strategy is also applicable for different HPV 16 E7 mutants and for E7 proteins from other HPV types such as HPV 18 and 11. PMID- 9226717 TI - Overexpression and purification of elongation factor 3 from Saccharomyces cerevisiae. AB - The translational elongation factor 3 (EF-3) from Saccharomyces cerevisiae was overexpressed and purified to near homogeneity from the post-ribosomal supernatant fraction (S-100). A detailed protocol for the isolation of overexpressed EF-3 is presented. The procedure involves ion-exchange chromatography on DEAE-Sepharose and CM-Sepharose and affinity chromatography on ATP-agarose. A protein purity of > or = 96% was established by quantitating the silver-stained SDS/polyacrylamide gel. The present method facilitates isolation of EF-3 in large amounts in high yield. PMID- 9226718 TI - Stable expression and purification of a secreted human recombinant prethrombin-2 and its activation to thrombin. AB - A human prothrombin cDNA has been engineered to obtain a cDNA coding for a secreted form of human prethrombin-2. The secreted prethrombin-2 has been produced in a mammalian expression system using DXB11 cells, a mutant strain of CHO cells in which the dihydrofolate reductase gene has been deleted, and an expression vector carrying the dihydrofolate reductase cDNA. Methotrexate-induced gene amplification favored an efficient production of the recombinant protein which accumulated in the culture medium of the DXB11 cells. Growth in suspension of the stable transformants in an airlift fermenter resulted in the production of 25 mg/L recombinant prethrombin-2. The recombinant protein was purified using single-step affinity chromatography on a recombinant-hirudin column and activated by agarose gel-immobilized ecarin. All purified recombinant prethrombin-2 was activated and the generated recombinant thrombin showed catalytic properties identical to those of plasma-derived alpha-thrombin. This expression system can be used to prepare mutants of prethrombin-2 for structure-function studies investigating thrombin interactions with substrate proteins, inhibitors, and cell membranes. PMID- 9226719 TI - Expression and purification of recombinant human apolipoprotein A-I in Chinese hamster ovary cells. AB - The expression of apolipoprotein A-I (apoA-I) has been shown to be very difficult due to its amphiphilic character, autoaggregation, and degradation. We have expressed apoA-I using CHO cells, Baculovirus, and Escherichia coli [Schmidt et al., J. Biol. Chem. (1995) 270, 469-475]. Here we report about optimized conditions for the expression of proapoA-I in CHO cells, testing various serum free media. We were able to yield apoA-I expression up to 80 micrograms/ml, by far the highest ever reported. However, immunoblot analysis revealed degraded apoA-I. The best apoA-I expression testing various conditions was about 20-30 micrograms/ml without any evidence of degradation. Interestingly, the apoA-I expression resulted in reproducible apoA-I fragments of 26 and 14 kDa. These fragments are consistent with already reported in vivo findings, in which carboxy terminal proteolysis was suggested. The use of the protease inhibitors pepstatin and chymostatin, both carboxy-peptidase inhibitors, did result in contrast to other studied protease inhibitors in increased apoA-I yield. Therefore, limited carboxy-terminal proteolysis contributes to the degradation of CHO cell-secreted apoA-I. In addition, we evaluated various purification methods for the preparative isolation of recombinant apoA-I. In our hands we obtained the best recovery and no degradation with reversed-phase chromatography using a FPLC system. PMID- 9226720 TI - Expression, purification, and functional analysis of the TyrR protein of Haemophilus influenzae. AB - The gene that was inferred to encode the TyrR protein of Haemophilus influenzae Rd was synthesized by polymerase chain reaction and inserted into a T7-based expression vector. Methods were developed to overexpress the TyrR protein of H. influenzae in Escherichia coli and to purify the protein on a large scale. Both in vitro and in vivo functional comparisons of the H. influenzae and E. coli TyrR proteins were carried out. The TyrR protein of H. influenzae was able to bind in vitro to an operator target upstream of the aroF-tyrA gene of E. coli. In the presence of [gamma-S]ATP, the DNA binding ability of the H. influenzae TyrR protein was drastically reduced. Despite the much shorter peptide chain length (318 amino acid residues vs 513), the TyrR protein of H. influenzae was as active in repressing the aroF promoter as the TyrR protein of E. coli. Repression by both proteins was enhanced in the presence of tyrosine; however, the transcriptional activation function associated with the TyrR protein of E. coli could not be detected when the H. influenzae TyrR protein was expressed in E. coli. By computer analysis, at least five operator targets for TyrR were identified within the genomic DNA of H. influenzae. These observations show that the assignment of function to the tyrR gene of H. influenzae was correctly made. Further studies of the H. influenzae TyrR protein in comparison to its E. coli counterpart should provide valuable mechanistic information on transcriptional regulation in this system. PMID- 9226721 TI - Conditions for the adsorption of proteins on ultrastable zeolite Y and its use in protein purification. AB - The adsorption of proteins on ultrastable zeolites was investigated. Protein binding to one of these, ultrastable zeolite Y (USY), was studied in detail. Protein binding to USY, with a Si/Al ratio of > 240, was found to be dependent on the pH of the solution, being highest at or just below the pI of the protein. The amount of protein adsorbed on the zeolite was found to be 10 times as much as the estimated binding to the external surface of the USY. We propose an adsorption mechanism involving the formation of a protein layer strongly bound to the USY surface, further protein layers being formed on top of this on the basis of protein-protein interactions. The protein-protein interactions can be disrupted by changing the pH. Ultrastable zeolite Y was used as a new matrix for protein purification. Undesired proteins can be removed from a crude preparation by adsorption on USY, increasing the purity of a specific protein, or the protein can be adsorbed on the zeolite and subsequently eluted through changing the pH. These two means of protein purification are exemplified by the purification of peroxidase from a crude horseradish extract and by the purification of lysozyme from egg white. PMID- 9226722 TI - High-level expression of human thymidylate synthase. AB - A method is presented for expressing human thymidylate synthase (TS) to the extent of 25-30% of the protein in Escherichia coli. By this procedure, 200-400 mg of pure enzyme can be obtained from a 2-liter culture of cells. The key to the level of expression appears to be related to the conversion of purine bases in the third, fourth, and fifth codons of the TS cDNA to thymine, without altering the encoded protein product. Conversion of the penultimate proline to a leucine did not diminish expression, but while the isolated native enzyme contained only proline on its amino-terminal end, the mutated enzyme was found to contain methionine on its amino terminus. By contrast, the expression of the unmodified TS cDNA represented only about 0.1-0.2% of the total cellular protein. Unlike recombinant rat and human TSs, the respective enzymes purified to homogeneity from eukaryotic cells were blocked at the amino ends and possessed 2- to 4-fold lower specific activities. To determine at what level the impairment of expression occurred, an in vitro transcription, translation system was employed and the results showed that while transcription was unaffected, the translation of native TS mRNA was reduced by at least 20-fold relative to modified TS mRNA using a rabbit reticulocyte translation system. Thus, it appears that at least for the TS gene, expression is greatly influenced by the GC content of the 5' coding region of the gene in both prokaryote and eukaryote systems. PMID- 9226723 TI - Expression, purification, and characterization of an activated cytokine suppressive anti-inflammatory drug-binding protein 2 (CSBP2) kinase from baculovirus-infected insect cells. AB - An activated form of the human cytokine-suppressive anti-inflammatory drug binding protein 2 (CSBP2) kinase was expressed in Spodoptera frugiperda (SF9) cells from a baculovirus vector. To maximize expression and to facilitate purification of the recombinant protein, CSBP2 kinase was expressed as a carboxy terminal fusion protein to glutathione S-transferase (GST). Under optimal conditions, 2-3 mg of GST-CSBP2 could be obtained per liter of infected cell culture. The fusion protein was easily purified from the soluble fraction of the total cell lysate under nondenaturing conditions by using a glutathione-Sepharose 4B affinity resin. As expected, the purified GST-CSBP2 fusion protein was approximately 68 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and reacted with antibodies directed toward either the GST or the CSBP amino terminus. To obtain activated CSBP2, SF9 cells were coinfected with two recombinant baculovirus vectors: one that directed the synthesis of the GST-CSBP2 fusion protein and a second vector that directed the synthesis of a constitutively active form of the CSBP activating kinase, MKK3. Coexpression of GST-CSBP2 kinase with the MKK3 activator increased GST-CSBP2 activity 8- to 10-fold based on the ability of GST-CSBP2 to phosphorylate the substrate, myelin basic protein (MBP), and the ATF2 transcription factor, in vitro. Moreover, activated GST-CSBP2 was capable of activating a bacterially derived mitogen-activated protein kinase-activating protein kinase 2 in vitro. The activity of insect-derived GST-CSBP2 was also inhibited by the CSBP inhibitor, SB202190. We anticipate that the preparation and purification techniques described in this study will facilitate further biochemical characterization of this kinase. PMID- 9226724 TI - Purification of UhpT, the sugar phosphate transporter of Escherichia coli. AB - To purify UhpT, the sugar phosphate carrier of Escherichia coli, we constructed a variant (HisUhpT) in which 10 tandem histidine residues were placed at the UhpT N terminus and then used Ni(2+)-agarose affinity chromatography of detergent solubilized proteins. Membrane vesicles from a strain overexpressing His-UhpT were extracted at pH 7.4 with either 1.5% n-octyl-beta-D-glucopyranoside (octylglucoside) or 1.5% n-dodecyl-beta-D-maltoside (dodecylmaltoside) in 200 mM sodium chloride, 100 mM potassium phosphate, 50 mM glucose 6-phosphate, 10-20% glycerol, 0.2% E. coli phospholipid, and 5 mM beta-mercaptoethanol. After the detergent extract was applied to a Ni(2+)-agarose column, nonspecifically bound material was removed by washing at pH 7 with the same buffer also containing 50 mM imidazole. Purified HisUhpT was released subsequently, when sodium chloride was replaced with 300 mM imidazole or 100 mM EDTA, giving an overall yield of about 25 micrograms HisUhpT/mg vesicle protein. Whether eluted by imidazole or EDTA in either octylglucoside or dodecylmaltoside, purified HisUhpT showed a specific activity of 2.5-3 mumol/min per milligram of protein as monitored by [14C]glucose 6-phosphate transport by proteoliposomes loaded with 100 mM potassium phosphate. This corresponded to a calculated turnover number near 20 s 1 for the heterologous exchange of external sugar phosphate with internal phosphate. At low temperature (4 degrees C) HisUhpT retained full activity in either octylglucoside or dodecylmaltoside; however, at elevated temperature (> or = 23 degrees C), the protein displayed a marked lability in octylglucoside (t1/2 = 11 min), but not in dodecylmaltoside (t1/2 > or = 200-300 min). PMID- 9226725 TI - Expression of human amyloid precursor protein ectodomains in Pichia pastoris: analysis of culture conditions, purification, and characterization. AB - We have examined the use of the yeast Pichia pastoris for expression of the human amyloid precursor protein (APP). The ectodomains of the isoforms APP695, APP751, and APP770 were expressed in both P. pastoris protease-deficient strain SMD1163 and wild-type strain GS115, using the secretion vector pHIL-S1. Expression of recombinant APP in each of these strains produced intact recombinant protein, together with a small number of breakdown products. The levels of these breakdown products were not significantly altered by expression in the protease-deficient strain compared with wild-type GS115. The effects of induction time and medium composition on recombinant APP stability were also examined. After optimization of expression and culture conditions, baffled shaker flask cultures of clones selected for high expression routinely yielded 13-24 mg/liter recombinant protein following a two-step purification procedure. The recombinant isoforms possessed the heparin binding, metal binding, and Kunitz-type protease inhibitor properties of human brain-derived APP. These data indicate that P. pastoris is an appropriate laboratory-scale expression system for production of sufficient quantities of recombinant APP for use in biological studies. PMID- 9226726 TI - Anti-sexual and anxiogenic behavioral consequences of corticotropin-releasing factor overexpression are centrally mediated. AB - Corticotropin-releasing factor (CRF) acts as a neurotransmitter in brain to promote behavioral responses such as flight and immobility, which have adaptive value in the context of exposure to environmental stressors. CRF also suppresses behavioral repertoires such as mating, which are incompatible with such threat related coping responses. In this study, we employed transgenic (Tg) mice which overexpress CRF in brain and exhibit a constitutive and persistent phenotype of emotionality in order to determine the consequences of long-term CRF excess on indices of reproductive success, male sexual performance and female sexual receptivity. Sexual performance of CRF Tg males was relatively intact, whereas female receptivity was masked in CRF Tg mice by active rejection of sexually experienced male counterparts. This impairment in social interaction was only partially normalized by the serotonin antagonist, methysergide, which enhanced olfactory exploration of the still non-receptive CRF Tg females. Moreover, the anxiogenic-like character of CRF Tg mice is likely to be centrally mediated, since attenuation of hypercorticosteronemia by adrenalectomy did not alter either impaired sexual receptivity or fear-like behavior in an animal model of anxiety. Thus, overexpression of CRF in the brain results in a variety of adverse consequences including diminished social interactions. PMID- 9226727 TI - Neuroendocrine measures and lymphocyte subsets in depressive illness: influence of a clinical interview concerning life experiences. AB - The effects of a clinical interview concerning either positive or negative day-to day events on lymphocyte subpopulations, and on plasma cortisol, ACTH and norepinephrine, were determined in depressive patients (major depressive and dysthymic) and in normal controls. Irrespective of its content, the interview provoked an elevation of circulating natural killer (NK) cells, suggesting that this effect was related to either a change in mood state (regardless of its valence) or to the stress associated with the interview procedure. Since the interview did not influence plasma cortisol, ACTH or norepinephrine, it is likely that the NK cell variations were independent of these endocrines. Although basal NK cells were elevated in the depressive group relative to controls, the extent of the NK cell increase provoked by the interview was comparable in depressive and control subjects. The failure to detect differences between these populations could not be attributed to ceiling effects precluding more pronounced alterations in the depressed subjects. Indeed, variations of circulating cell subtypes were found to be exquisitely sensitive to differences in stressor intensity. In a subset of control subjects, a more potent stressor (anticipation of an academic examination) increased the plasma endocrine levels, increased circulating NK cell number beyond that associated with the interview stress, and provoked an increase of several T cell subsets (CD3, CD4 and CD8). Evidently, while a clinical interview may be sufficiently stressful to influence circulating NK cells, the stress of such a procedure seems no greater in depressed than in control subjects. It is suggested that although depressed patients may exhibit higher basal NK levels, this effect is likely not related to increased reactivity to stressors. PMID- 9226729 TI - Blunted serotonin-mediated activation of the hypothalamic-pituitary-adrenal axis in chronic fatigue syndrome. AB - We examined 5HT1a-mediated ACTH release in patients with chronic fatigue syndrome (CFS) using a between-subjects design. Patients attending a specialist outpatient clinic for CFS, who fulfilled CDC criteria, together with age- and sex-matched healthy comparison subjects, were recruited. Subjects had a cannula inserted in a forearm vein at 0830 h and were allowed to relax until 0900 h, when baseline bloods for ACTH and cortisol were drawn. They were then given ipsapirone 20 mg PO and further blood for hormone estimation was taken at +30, +60, +90, +120 and +180 min. Baseline ACTH and cortisol levels did not differ between the two groups. Release of ACTH (but not cortisol) in response to ipsapirone challenge was significantly blunted in patients with CFS. We conclude that serotonergic activation of the hypothalamic-pituitary-adrenal axis is defective in CFS. This defect may be of pathophysiological significance. PMID- 9226728 TI - A neuroendocrine role in cocaine reinforcement. AB - Cocaine stimulates the secretion of corticosterone and ACTH, probably through a CRF-related mechanism, indicating that the drug activates the HPA axis. Indeed, cocaine has been reported to produce anxiety and to precipitate episodes of panic attack during chronic use and withdrawal in humans and to induce anxiogenic behavior in animals. Cocaine also alters benzodiazepine receptor binding in discrete regions of the rat brain. Some of these changes in binding are obviously related to the convulsions and seizures which are often observed in an acute cocaine overdose. However, data from behavioral studies have suggested that some of these effects may be related directly to cocaine reinforcement since receptor changes also were observed when binding in the brains of rats that self administered cocaine was compared with that from animals that had received identical yoked, but non-contingent infusions of the drug. In this regard, pretreatment with the benzodiazepine receptor agonists chlordiazepoxide and alprazolam decreased cocaine self-administration without decreasing food reinforced responding, suggesting that these effects were specific for cocaine. Since this attenuation of self-administration was reversed by increasing the unit dose of cocaine, it is likely that these drugs were decreasing cocaine reinforcement. In contrast, exposure to stress increases vulnerability to self administer psychostimulants. In these experiments, low-dose cocaine self administration was related directly to stress-induced increases in plasma corticosterone, such that plasma corticosterone was always greater than 150 ng/ml for rats which subsequently self-administered cocaine at doses of 0.125 mg/kg/infusion or lower, suggesting a threshold for the hormone in cocaine reinforcement. In other experiments, bilateral adrenalectomy completely abolished the acquisition of intravenous cocaine self-administration in naive rats, while metyrapone decreased ongoing self-administration. In addition, ketoconazole pretreatment resulted in patterns of self-administration that were virtually indistinguishable from that observed during saline extinction, suggesting that plasma corticosterone is not only important, but may even be necessary for cocaine reinforcement. The mechanisms through which adrenocorticosteroids alter cocaine reinforcement remain to be determined, but there is increasing evidence that the mesocorticolimbic dopaminergic system is involved. In particular, the medial prefrontal cortex appears to be at least one brain region where dopamine and adrenocorticosteroids may interact to affect cocaine reinforcement. PMID- 9226730 TI - Nullification of a positive correlation between urinary levels of alpha 1 microglobulin and ulinastatin by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) in mice. AB - The relationships between urinary levels of alpha 1-microglobulin (alpha 1M) and ulinastatin (UT) were investigated in C57BL/6J mice, a species which reportedly possesses the gene similar to that of humans for synthesizing the precursor protein of alpha 1M and UT. A positive correlation was established in normal mice. However, repetitive administrations (20 mg/kg, IP, four administrations/12 h) of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) nullified the positive correlation. A similar phenomenon was induced by ICV-administered MPTP (18 and 36 micrograms) in the animals. Furthermore, L-dopa administration (50 mg/kg, IV) in MPTP-treated (1 week after the final IP administration of MPTP) mice reversed the tendency of MPTP, although the agent alone did not affect the positive correlation in normal mice. These results suggest that nullification of the positive correlation probably was induced by the central effects of MPTP. We have found previously that the lack of a positive correlation between urinary levels of alpha 1M and UT distinguishes Parkinson's disease from other neuropsychiatric diseases such as dementia (Alzheimer-type and vascular dementia), schizophrenia and mood disorders. Our present results displayed a phenomenon that the lack of correlation between urinary levels of alpha 1M and UT in patients with Parkinson's disease is reproducible in MPTP-treated mice. PMID- 9226731 TI - Effects of the Transcendental Meditation program on adaptive mechanisms: changes in hormone levels and responses to stress after 4 months of practice. AB - Stress has been implicated in both somatic and mental disorders. The mechanisms by which stress leads to poor health are largely unknown. However, studies in animals suggest that chronic stress causes high basal cortisol and low cortisol response to acute stressors and that such changes may contribute to disease. Previous studies of the Transcendental Meditation (TM) technique as a possible means of countering effects of stress have reported altered levels of several hormones both during the practice and longitudinally after regular practice of this technique. In this prospective, random assignment study, changes in baseline levels and acute responses to laboratory stressors were examined for four hormones-cortisol, growth hormone, thyroid-stimulating hormone and testosterone before and after 4 months of either the TM technique or a stress education control condition. At pre- and post-test, blood was withdrawn continuously through an indwelling catheter, and plasma or serum samples were frozen for later analysis by radioimmunoassay. The results showed significantly different changes for the two groups, or trends toward significance, for each hormone over the 4 months. In the TM group, but not in the controls, basal cortisol level and average cortisol across the stress session decreased from pre- to post-test. Cortisol responsiveness to stressors, however, increased in the TM group compared to controls. The baselines and/or stress responsiveness for TSH and GH changed in opposite directions for the groups, as did the testosterone baseline. Overall, the cortisol and testosterone results appear to support previous data suggesting that repeated practice of the TM technique reverses effects of chronic stress significant for health. The observed group difference in the change of GH regulation may derive from the cortisol differences, while the TSH results are not related easily to earlier findings on the effects of chronic stress. PMID- 9226732 TI - Modulating effects of a cold water stimulus on opioid effects in volunteers. AB - The purpose of this study was to characterize the effect of a painful stimulus on morphine and butorphanol effects in healthy non-drug abusing volunteers. Thirteen subjects with no history of opiate dependence participated in a randomized, placebo-controlled, double-blind, crossover trial in which each subject received saline, 2 mg/70 kg butorphanol, and 10 mg/70 kg morphine, IV, in each of two conditions, periodic forearm immersions into either ice-cold water (2 degrees C) or into warm water (37 degrees C). Both opioids reduced self-reported ratings of pain intensity, indicative of analgesia. Several of the subjective effects of morphine were attenuated either during or in between cold-water immersions, including visual analog scale ratings of "coasting (spaced out)," "high (drug "high")," "sleepy (drowsy, tired)," and "lightheaded". In contrast, some of butorphanol's subjective effects were increased by the cold-water manipulation. Morphine impaired psychomotor performance during one of the warm-water immersions, but not during the cold-water immersions. Psychomotor impairment induced by butorphanol was not affected by water temperature. This study provides evidence that opioid effects can be modulated by a painful stimulus in humans. PMID- 9226733 TI - Effect of imipramine treatments on the 5-HT1A-receptor-mediated inhibition of panic-like behaviours in rats. AB - The escape behaviour induced in rats by injecting D,L-homocysteic acid (DHL) into the dorsal periaqueductal grey area (DPAG) was used as an animal model of panic attacks to investigate the effect of imipramine, a drug used for the treatment of panic disorder, on the sensitivity of 5-HT1A receptors in the DPAG. Rats given imipramine (10 mg/kg per day SC for 3 weeks or IP for 2-3 days) received 250 nl saline or the 5-HT1A agonist 8-OH-DPAT (8.6 nmol) into the DPAG 10 min before inducing the escape response with DLH. As expected, 8-OH-DPAT produced a marked decrease in the average speed of the DLH-induced flight response. The short-term treatment with imipramine changed neither the DLH-induced escape behaviour nor the effect of prior 8-OH-DPAT administration on this response. In contrast, long term treatment with imipramine enhanced the 5-HT1A-mediated inhibition, as the decrement in the amplitude of the flight response produced by 8-OH-DPAT was 96% after this treatment compared to 41% in controls. The injection of 8-OH-DPAT also significantly decreased the amplitude of the freezing behaviour observed at the end of the flight response in rats given imipramine for 3 weeks, but not in controls. The long-term imipramine treatment, however, did not significantly decrease the amplitude of DLH-induced flight and freezing behaviours in absence of prior 8-OH-DPAT administration. Finally, 8-OH-DPAT failed to inhibit the DLH induced flight and freezing behaviours in rats withdrawn from imipramine after long-term treatment (10 mg/kg per day x 21 days). It is suggested that an alteration at the level of the DPAG-5-HT1A receptor system is implicated in the therapeutic and withdrawal effect of imipramine in panic disorder. PMID- 9226734 TI - Effects of haloperidol and amisulpride on motor and cognitive skill learning in healthy volunteers. AB - The effects of a typical neuroleptic, haloperidol (1 and 2 mg orally), of an atypical neuroleptic, amisulpride (50 and 100 mg) and of a placebo on motor and cognitive skill learning were assessed in 60 healthy volunteers using repeated testing on the Tower of Toronto puzzle. Subjects were asked to solve three blocks of eight trials and, at distance from drug administration, a fourth block. The puzzle was connected to a computer in order to obtain a precise timing of individual moves. Two components of cognitive skill learning were assessed, the ability to learn to solve the puzzle and the acquisition of a problem-solving routine. Subjective feelings of effort and automatisation of the task were assessed using a questionnaire. Like placebo-treated subjects, neuroleptic treated subjects were able to acquire a motor skill, to learn to solve the puzzle and to acquire a routine. However, haloperidol 2 mg-treated subjects needed significantly more moves to solve the puzzle in blocks 3 and 4, some of them having routinised a non-optimal solution. A significant cognitive slowing was observed in the haloperidol 1 mg group in block 4. The performance pattern and verbal reports suggested that haloperidol impaired the higher cognitive functions such as the ability to shift from one strategy to another and/or to assess one's performance accurately, possibly leading to the development of compensatory strategies. The only deleterious amisulpride effect was a cognitive slowing in block 4, which was observed in the lower dose group. PMID- 9226735 TI - Central 5-HT2A and D2 dopamine receptor occupancy after sublingual administration of ORG 5222 in healthy men. AB - It has been suggested that a combined blockade of 5-HT2A and D2-dopamine receptors improves efficacy and reduces the risk for extrapyramidal symptoms when treating schizophrenic patients with antipsychotic drugs. ORG 5222 is a new potential antipsychotic drug which has high affinity for 5-HT2A, D2-dopamine and alpha 1 adrenergic receptors in vitro. The objective of this study was to examine if ORG 5222 occupies 5-HT2A and D2-dopamine receptors in human subjects in vivo. Central receptor occupancy was measured by positron emission tomography (PET) in three healthy subjects after sublingual administration of 100 micrograms ORG 5222. [11C]N-methylspiperone ([11C] NMSP) was the radioligand used to measure 5 HT2A receptor binding in the neocortex and [11C]raclopride to measure D2-dopamine receptor binding in the striatum. The 5-HT2A occupancy was 15-30% and the D2 dopamine receptor occupancy was 12-23%. The study confirms that ORG 5222 binds to 5-HT2A and D2-dopamine receptors in human brain. Since receptor occupancy of ORG 5222 is rather low, doses higher than 100 micrograms are suggested in future clinical trials to evaluate the antipsychotic drug effect of ORG 5222. PMID- 9226736 TI - Interactions between H1-antagonists and opioids: a drug discrimination study. AB - We previously demonstrated that combination of opioids, pentazocine and dihydrocodeine, with the histamine H1-receptor antagonists tripelennamine and chlorpheniramine could enhance their rewarding effects in rats. In the present study, the effects of combined treatment with opioids and H1-antagonists on discriminative stimulus effects were examined in rats trained to discriminate between cocaine (10 mg/kg) or morphine (3.0 mg/kg) and saline, since it is believed that discriminative stimulus effects of abused drugs are related to their rewarding effects. Tripelennamine and chlorpheniramine, but not pentazocine or dihydrocodeine, generalized to the discriminative stimulus effects of cocaine. Pentazocine (3.0 mg/kg) and dihydrocodeine (5.6 mg/kg) significantly potentiated the cocaine-like discriminative stimulus effects of low doses of tripelennamine and chlorpheniramine, respectively. On the other hand, pentazocine and dihydrocodeine, but not tripelennamine or chlorpheniramine, generalized to the discriminative stimulus effects of morphine. Neither 1.0 or 3.0 mg/kg tripelennamine nor chlorpheniramine affected the morphine-like discriminative stimulus effects of pentazocine and dihydrocodeine, respectively. These results suggest that the potentiation of the cocaine-like discriminative stimulus effects of H1-antagonists by opioids may be involved in the enhanced rewarding effects of combinations of opioids and H1-antagonists. PMID- 9226737 TI - Flumazenil blockade of anxiety following ethanol withdrawal in rats. AB - In previous research, the drug flumazenil has been categorized both as a pure benzodiazepine antagonist and as a benzodiazepine partial agonist. The following studies used an elevated plus maze to test whether flumazenil would exert any antianxiety action in rats. While chlordiazepoxide (3.0 mg/kg), ethanol (0.75 g/kg), and the atypical benzodiazepine zolpidem (1.0 mg/kg) all significantly increased time spent on the open arms and percent open arm entries, flumazenil (1 10 mg/kg) alone did not produce any anxiolytic effects on the maze. Withdrawal from chronic ethanol treatment led to a decrease in open arm time and percent open arm entries. Flumazenil (3.0 mg/kg) blocked these changes, suggesting that the effects of flumaxenil are at least partially dependent upon the levels of stress or anxiety in the subjects. An anxiolytic action of flumazenil was not seen following the central administration of the neuropeptide corticotropin releasing factor (CRF), which reduced open arm time on the elevated plus maze. These results support the hypothesis that the mechanism of action for flumazenil effects on the anxiety observed during ethanol withdrawal involves antagonism of an endogenous benzodiazepine inverse agonist, rather than activity as a partial agonist or blockade of CRF-mediated effects. PMID- 9226738 TI - Nicotine discrimination and self-administration in humans as a function of smoking status. AB - Nicotine's discriminative stimulus effects may be critical to understanding reinforcement of tobacco smoking. It is not known whether regular nicotine exposure produces tolerance or sensitivity to these effects. In this study, male and female smokers (n = 11) and never-smokers (n = 10) were trained to discriminate 20 micrograms/kg nicotine by nasal spray from placebo (0) on day 1. On day 2, both groups were tested on generalization of this discrimination across intermittent presentations of 0, 3, 6, 12, and 20 micrograms/kg nicotine in random order. Quantitative and quantal behavioral discrimination tasks, used in previous research, were employed. On day 3, subjects were instructed to self administer sprays from the 20 micrograms/kg nicotine versus 0 bottles in a concurrent-choice procedure. All but one subject (female smoker) learned reliably to discriminate 20 micrograms/kg nicotine from placebo (> or = 80% correct) on day 1. Nicotine-appropriate responding on day 2 was attenuated in smokers versus never-smokers at 20 micrograms/kg on the quantitative task and at 12 micrograms/kg on the quantal task, suggesting tolerance. There was no difference in responding at other doses. Smokers also showed attenuated responses on the subjective measure of "head rush", which was associated with discrimination responding in both groups. Nicotine self-administration was significantly greater in smokers versus never-smokers, who self-administered nicotine below chance levels, and was inversely related to discrimination behavior in never-smokers but unrelated in smokers. Women smokers showed less change in nicotine-appropriate responding across generalization doses, reported less confidence in discriminating training doses during acquisition on day 1, and tended to self administer less nicotine on day 3. These results indicate that smokers may become tolerant to the discriminative stimulus effects of nicotine, perhaps promoting increased use. PMID- 9226739 TI - Executive function and uptake of 99mTc-exametazime shown by single photon emission tomography after oral idazoxan in probable Alzheimer-type dementia. AB - Preliminary reports suggest improved executive function in patients with lobar dementia after treatment with single doses of the alpha 2 adrenoceptor antagonist, idazoxan. The potential for use in probable Alzheimer-type dementia prompted the present study. Fifteen patients with probable Alzheimer-type dementia were examined twice with neuropsychological measures and 14 also with single photon emission tomography (SPET) after a single double blind oral administration of 40 mg idazoxan or placebo in a balanced cross-over design. Brain perfusion maps were spatially transformed into standard stereotactic space and compared pixel-by-pixel. A parametric analysis was used to examine the relationship between the drug effect, verbal fluency and brain perfusion. Two to 3 h after idazoxan, measures of reaction time, Stroop test, category fluency and anxiety were unchanged. Verbal fluency (letter) and spatial working memory were impaired and performance on the Tower of London test in a sub-set of patients showed a trend to impairment in the idazoxan condition. Idazoxan produced a modest relative activation in left thalamus and inferior occipital cortex: decreases occurred in inferior anterior cingulate and left insular cortex. There were significant correlations on both days between measures of fluency and brain perfusion in left lateral prefrontal cortex. The reduced performance with idazoxan was directly correlated with reduced perfusion in left lateral prefrontal cortex, supporting an important interaction between drug and task performance. The imaging component of the study therefore suggested that activation of frontal networks is necessary for performing fluency tasks in Alzheimer-type dementia. Brain networks involving prefrontal cortex are the locus for the primary cognitive effects of noradrenergic drugs. The direction of the effect of any dose of agonist or antagonist may depend critically upon the age and pathology of the experimental subjects and the relationship between performance, noradrenergic drive and task difficulty. PMID- 9226740 TI - Cortical and hippocampal EEG power spectra in animal models of schizophrenia produced with methamphetamine, cocaine, and phencyclidine. AB - Cortical and hippocampal EEGs in animal models of schizophrenia were compared to those obtained with psychotomimetics or antipsychotic agents by utilizing power spectral analysis. Models of positive schizophrenic symptoms were created with methamphetamine (MAP) and cocaine, and a model of both negative and positive symptoms was created with PCP. MAP caused a prolonged decrease in the cortical EEG power spectra, cocaine caused a marked decrease for a short time, and PCP produced no significant changes. In the hippocampal spectra, MAP induced a marked increase in the T2(6.0-7.9 Hz)/ T1(4.0-5.9 Hz) ratio, PCP caused a decrease of this ratio after an initial increase, and cocaine produced no significant change. (An increase in the T2/T1 ratio represents a shift of theta waves to higher frequencies.) Since apomorphine (a DA agonist) and MK-801 (an NMDA antagonist) caused the T2/T1 ratio to increase, positive schizophrenic symptoms caused by MAP may be related to the DA and NMDA systems. 3-PPP (a sigma agonist) caused biphasic changes similar to those induced by PCP. Haloperidol and chlorpromazine caused a decrease of the T2/T1 ratio. These results indicate that cortical and hippocampal EEG power spectra (especially the hippocampal T2/T1 ratio) can be used to characterize both qualitatively and quantitatively models of schizophrenia. PMID- 9226741 TI - Intracerebral penetration of carteolol hydrochloride in rats. AB - To elucidate the penetrability of carteolol, a beta-adrenoceptor antagonist (beta blocker) into the brain of rats, intracerebral and serum concentrations of the compound were determined in male rats receiving single or repetitive oral administration of carteolol hydrochloride at 30 mg/kg. The time-course of the intracerebral concentration of carteolol following single IV administration of the compound at 10 and 30 mg/kg was also studied in male rats. A high-performance liquid chromatography method was used to determine the intracerebral and serum concentrations. Following single oral dosing, the intracerebral concentration of carteolol reached a maximum of 0.074 microgram/g at 2 h postdosing and declined with a half-life of 3.7 h, and the Cmax and AUC of carteolol in the brain were 12.5% and 19.8% of those in serum. The intracerebral and serum concentrations of carteolol were determined in male rats receiving repetitive oral dosing of the compound once daily for 7 days. The concentration of carteolol in the brain and serum at 1 h postdosing varied within a range of 0.059-0.091 microgram/g and 0.321-0.443 microgram/ml, respectively, throughout the dosing period, showing no changes in the penetrability of the compound into the brain due to repeated dosing. The concentration of carteolol in the brain and serum increased in a dose dependent manner in rats receiving a single IV administration of the compound. The elimination half-life of carteolol in the serum and brain was 0.6-0.8 h and 1.3-1.7 h, respectively, in rats following single IV dosing of the compound. The half-life in the brain was about twice as long as that in the serum. The brain to serum concentration ratio was 0.306:0.499. From the above results, it was concluded that carteolol is distributed from the circulation to the brain with low penetrability. PMID- 9226742 TI - Differences in enhancing effects of zolpidem and benzodiazepine drugs on recurrent inhibition in rat hippocampal slices. AB - It has been reported that the clinical and electroencephalographic profiles of zolpidem, a non-benzodiazepine drug which binds preferentially to the omega 1 benzodiazepine recognition sites located within the GABAA receptor complex, are different from those of benzodiazepine drugs, which bind non-selectively to the omega 1 and omega 2 sites. In order to clarify the electrophysiological mechanism underlying the unique profile of zolpidem, the present study compared the enhancing effects of zolpidem and two benzodiazepine drugs, triazolam and diazepam, on recurrent inhibition. This inhibition was expressed as suppression of the orthodromically induced population spikes by the preceding antidromic stimulation of the alveus in the CA1 region of rat hippocampal slices. The rank order of potency for enhancing recurrent inhibition was triazolam > diazepam > zolpidem. From the present results and previously reported findings that zolpidem has a lower affinity for the omega 2 sites than diazepam while both have the same affinity for the omega 1 sites, we concluded that the hippocampal recurrent inhibition appears to be enhanced mainly by activation of the omega 2 sites, but not by that of the omega 1 sites. Furthermore, the lower potency of zolpidem for enhancing recurrent inhibition may underlie its unique profile in terms of its clinical and electroencephalographic effects. PMID- 9226743 TI - 7-Nitroindazole, a nitric oxide synthase inhibitor, has anxiolytic-like properties in exploratory models of anxiety. AB - The action of the novel nitric oxide synthase (NOS) inhibitor 7-nitroindazole (7 NI) was studied in different exploratory models of anxiety. In the rat plus-maze test, 7-NI potently increased time spent on open arms and percentage of open arm visits in a dose dependent manner with the minimal effective dose of 40 mg/kg. 7 NI caused an anxiolytic-like effect in the rat social interaction test. The minimal dose increasing social interaction time was 20 mg/kg. However, the drug also produced a clear sedative effect occurring even at smaller doses (10 mg/kg) in the open field test. 7-NI also showed an anxiolytic-like profile in the mouse light-dark compartment test and in the elevated plus-maze test, but the doses required were higher (80-120 mg/kg) than in rat models. Also, the sedative effect occurred at these doses in open field. We failed to demonstrate any effect of L arginine either in the rat elevated plus-maze test or in the open field test at doses up to 600 mg/kg IP. These results indicate that there are no major interspecies differences between rats and mice in respect of action of 7-NI. The clear anxiolytic-like action of the nitric oxide synthase inhibitor in four different models shows that nitric oxide is involved in the process of anxiety and that NOS could be a new target in developing anxiolytic drugs. PMID- 9226744 TI - Enhanced performance of spatial and visual recognition memory tasks by the selective acetylcholinesterase inhibitor E2020 in rhesus monkeys. AB - Hepatotoxicity limits the clinical utility of the cholinesterase inhibitor tacrine as a palliative therapy for Alzheimer's disease. The present studies examined the effects of E2020, a selective acetylcholinesterase inhibitor not associated with liver toxicity in man, on cognitive performance in rhesus monkeys using tasks employed previously to evaluate tacrine and other cholinomimetic agents. The ability of E2020 to prevent the induction of a cognitive impairment by the muscarinic receptor antagonist scopolamine was assessed using an automated spatial delayed response task. Coadministration of E2020 (0.5-1.75 mg/kg) caused a dose-dependent reversal of the scopolamine (0.03 mg/kg) induced impairment observed after retention intervals of 10 and 20 s. At the highest dose of E2020 examined (1.75 mg/kg), choice accuracy approached normal control levels. In this dose range, E2020 was well tolerated, but at the higher dose of 2 mg/kg, cholinergic side-effects were apparent. The effect of E2020 on choice accuracy in a visual recognition task was also assessed as this task does not require the use of scopolamine to disrupt performance and beneficial effects of cholinomimetics can therefore be detected at lower doses than in the spatial memory paradigm. In this task, administration of E2020 increased choice accuracy from 59 +/- 1% correct to up to 71 +/- 2% at doses of 0.03 and 0.05 mg/kg. No observable adverse effects were induced by E2020 in this dose range. The ability of E2020 to improve performance in these cognitive tasks resembles the profile of other cholinesterase inhibitors, including tacrine, that also improve cognitive function in Alzheimer's disease patients. Because of its more favourable clinical safety profile, E2020 may provide a significantly improved palliative therapy for dementia. PMID- 9226746 TI - Gliclazide scavenges hydroxyl, superoxide and nitric oxide radicals: an ESR study. AB - The role of reactive oxygen species in diabetes and its complications are well known. Two therapeutic agents commonly used in the treatment of diabetes are the sulfonylureas, gliclazide and glibenclamide. These drugs effectively reduce blood sugar in non-insulin dependent diabetes millitus by augmenting insulin release. Gliclazide is known to be a general free radical scavenger as demonstrated by inhibition of o-dianisidine photo-oxidation. In this study, the effects of gliclazide and glibenclamide on free radicals were examined in vitro, using electron spin resonance (ESR) spectroscopy. Superoxide radical (O2.-) generated from hypoxanthine-xanthine oxidase system, or hydroxyl radical (.OH) generated by the Fenton reaction, were analyzed as spin adducts of 5,5-dimethyl-1-pyrroline-N oxide (DMPO). NO was generated from 1-hydroxy-2-oxo-3-(N-3-methyl-3-aminopropyl) 3-methyl-1-triazene (NOC-7), and analyzed by 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl (carboxy-PTI) produced from the reaction between 2 (4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (carboxy-PTIO) and NO. Gliclazide scavenged O2.-, .OH and NO in a dose-dependent manner whereas glibenclamide was without effect. These findings suggest that gliclazide is not only effective in reducing blood sugar but also may be beneficial by inhibition of lipid and protein denaturation, which leads to the development of diabetic complications. PMID- 9226745 TI - Abnormal ACTH and prolactin responses to fenfluramine in rats exposed to single and multiple doses of MDMA. AB - The present study examined the persistent functional consequences associated with exposure to single and multiple doses of (+/-) 3,4-methylenedioxymethamphetamine (MDMA) as reflected by the neuroendocrine responses to d,l-fenfluramine (FEN). Adult male rats were administered a single dose of MDMA (20 mg/kg, s.c.) and challenged 2 weeks later with saline or FEN (2, 4, 6 and 8 mg/kg, s.c.). The corticotropin (ACTH) response to FEN (6 and 8 mg/kg) was blunted and the prolactin response to FEN (4 and 6 mg/kg) was enhanced in MDMA pre-treated rats. The ACTH and prolactin responses to FEN (6 mg/kg, s.c.) were then evaluated 4, 8 and 12 months after exposure to single and multiple doses MDMA (20 mg/kg, s.c. and 20 mg/kg, s.c., bid, x 4 days, respectively). The ACTH response to FEN was significantly reduced at 4 and 8 months in both MDMA treatment groups, and at 12 months in the multiple dose group only. In contrast, the prolactin response to FEN was enhanced in both groups of MDMA treated rats at 4 months, but only in the multiple dose group at 8 months. By 12 months, the prolactin response to FEN had normalized. Following multiple doses of MDMA, 5-HT concentrations were reduced significantly in the frontal cortex at 4 and 12 months. The results indicate that exposure to single or multiple doses of MDMA can produce functional alterations which can persist for months, whereas the biochemical sequelae were less robust and shorter lived. PMID- 9226747 TI - Free radical scavenging properties of alacepril metabolites and lisinopril. AB - Alacepril is an inhibitor of the angiotensin converting enzyme (ACE), and is commonly used as an antihypertensive. In this study, the effects of alacepril, its metabolites, desacetylalacepril and captopril, and also lisinopril, which has no sulfhydryl group in the structure, on free radicals were examined in vitro, using an ESR method. Superoxide and hydroxyl radical scavenging activities of alacepril metabolites, desacetylalacepril and captopril, were observed, whereas lisinopril hardly scavenged the superoxide or the hydroxyl radicals. Alacepril and its metabolites did not scavenge nitric oxide, but lisinopril showed slight scavenging activity. These findings suggest that the biological action of alacepril may be partly due to the antioxidant effect of its metabolites, having a sulfhydryl group. PMID- 9226748 TI - Production of ascorbate and hydroxyl radicals in the liver of LEC rats in relation to hepatitis. AB - The production of radicals was examined in vitro in liver supernatant prepared from LEC rats of different ages before and after the onset of jaundice. Each liver supernatant was subjected to heat-treatment at 90 degrees C for 10 min to remove heat-labile proteins, and then the production of radicals in the resultant supernatant in the presence of hydrogen peroxide and 5,5'-dimethyl-1-pyrroline oxide (DMPO) was studied by ESR. Two sharp ESR signals that completely decayed with time within 5 min after the addition of hydrogen peroxide were observed for the sample prepared from LEC rats before the onset of jaundice, followed by the appearance of four signals of a hydroxyl radical-DMPO adduct after 5 min. On the other hand, in the supernatant prepared from LEC rats after the onset of jaundice, the former two signals were not observed or observed only marginally, and the signals of the hydroxyl radical-DMPO adduct showed a different pattern of decay from that for the supernatant prepared from LEC rats before the onset of jaundice. With the addition of ascorbic acid to the liver supernatant prepared from LEC rats after the onset of jaundice, the former signals of the ascorbate and hydroxyl radicals reappeared. The present results suggest that ascorbate and hydroxyl radicals are produced in the liver of LEC rats with the onset of jaundice, depending on the relative ratio of ascorbic acid and cuprous ions. PMID- 9226749 TI - Characteristics of vasodilatation induced by cyclopiazonic acid in the rat perfused kidney. AB - We examined the possible existence of a novel endothelium-derived relaxing factor in the endothelium of the perfused rat kidney. Acetylcholine-induced vasodilatation was abolished by treatment with NG-nitro-L-arginine (L-NNA) and methylene blue in isotonic high K+ (60 mM) medium, whereas cyclopiazonic acid (CPA)-induced vasodilatation of the perfused kidney was slightly increased by this treatment. When the kidney was perfused with 0.47% CHAPS solution for 1 min, acetylcholine- or cyclopiazonic acid-induced vasodilatation was almost abolished. Acetylcholine- or cyclopiazonic acid-induced vasodilatation was not affected by indomethacin. These results suggest that CPA may release a novel endothelium derived relaxing factor, which is not prostanoids, nitric oxide nor endothelium derived hyperpolarizing factor in the endothelium of the renal vascular bed. PMID- 9226750 TI - Discrepancy between inotropic response and cyclic AMP production induced by isoprenaline and forskolin in rat ventricle strips. AB - The effects of isoprenaline and forskolin were studied on contractile response and cyclic AMP levels in the right ventricle strips of the rat heart. Isoprenaline, in concentrations ranging from 10 nM to 1 microM, significantly increased, in a concentration-dependent manner, the contractile force in this preparation. Forskolin (1-10 microM), which directly stimulates adenylate cyclase, also produced a concentration-dependent increase in cardiac contractility. The mean EC50 (microM) for the contractile action of isoprenaline was 0.08 +/- 0.014 and that of forskolin 7.3 +/- 1.1 being about 70 times less potent than isoprenaline. However, isoprenaline (0.1 microM) and forskolin (8 microM), which produced about the same inotropic response, increased tissue cyclic AMP levels by about two and five fold respectively, when compared to the basal value. These figures further indicate that while the intracellular levels of cyclic AMP in rat ventricular myocardium may be an important determinant of positive inotropism, the connection between the two parameters is more complex than the simple ratio between the tension generated and the amount of cyclic AMP found inside the cells. PMID- 9226751 TI - Inhibitory effects of Chinese ant extract (CAE) on nephrotoxicity induced by ferric-nitrilotriacetate (Fe-NTA) in Wistar rats. AB - We observed the inhibitory effects of Chinese ant extract (CAE), a Chinese traditional medicine, on nephrotoxicity induced by Fe-NTA in Wistar rat. Strong positive staining with Schiff's reagent was found in the proximal tubules of the untreated control rats. In contrast, the positivity was very weak in CAE treated rats. The level of TBARS was also higher in the untreated control rats than in CAE treated rats. Meanwhile, the scavenging effect of CAE on hydroxyl radicals was analyzed by electron spin resonance (ESR) in vitro. The results indicate that CAE can efficiently prevent Fe-NTA induced nephrotoxicity through quenching free radicals mechanism. PMID- 9226752 TI - Insulin secretion impairment and insulin sensitivity improvement in adult rats undernourished during early lactation. AB - In order to study the effect of undernutrition on the onset of disturbances in insulin secretion and insulin resistance, we compared the effects of a low protein diet containing 4% of protein (LPD) and a normal diet containing 25% of protein (NPD) supplied to the dams during the first 10 days of lactation when the pups turn into adults (90 days). We studies, in these rats, the insulin secretion, the glucose tolerance test (GTT) and, using the glucose clamp technique, the insulin resistance. The GTT showed a delay of the response of LPD group to the glucose challenge (1 mg/kg body weight) at 10 minutes (NPD = 450 +/- 27 mg/dl; LPD = 650 +/- 32 mg/dl, p < 0.01). The insulin secretion, four minutes after stimulation was found reduced in the LPD group (LPD = 1.1 +/- 0.08 microU/islet/min; NPD = 1.85 +/- 0.02 microU/islet/min, p < 0.01). Using the glucose clamp technique the plasma glucose concentration was raised during the first 20 minutes after the glucose stimulation with 10 mg/Kg-1.min-1 (NPD = 200 +/- 32 mg/dl and; LPD = 160 +/- 14 mg/dl., p < 0.01). Afterwards, the hyperglycemia was subsequently maintained (NPD = 154 +/- 9 mg/dl; LPD = 149 +/- 12 mg/dl) and the insulinemia was unchanged by infusion of glucose in the LPD group. In a similar experiment, the administration of glucose (10 mg/Kg-1. min-1) plus insulin (1.67 mU/Kg-1. min-1), the LPD group when compared with the NPD group, displayed an accentuated decreasing of glucose concentration level (LPD = 90 +/- 7 mg/dl; NPD = 130 +/- mg/dl., p < 0.01), 30 minutes after the infusion. The data suggest that undernutrition induces an adaptive process of insulin sensitivity which occurs together with an insulin secretion first phase blockage. PMID- 9226753 TI - Glycogenolysis stimulation by non-steroidal anti-inflammatories in the perfused rat liver is not accompanied by Ca2+ release. AB - Several non-steroidal anti-inflammatories increase glycogenolysis and are also able to affect intracellular Ca2+ channels. In the perfused liver, glycogenolysis stimulation is often associated with a transient Ca2+ efflux, which occurs immediately after the introduction of the glycogenolytic agonist. The purpose of this work was to verify if the introduction of non-steroidal anti-inflammatories also stimulates Ca2+ efflux. Rat livers were pre-loaded with 45Ca2+ in a recirculating system. The anti-inflammatory drugs diclofenac and niflumate were infused 10 minutes after restoration of the once-through perfusion. Glycogenolysis and oxygen uptake stimulation caused by these compounds was not accompanied by any increment in 45Ca2+ release. Subsequent infusion of vasopressin greatly stimulated 45Ca2+ efflux, denoting that the intracellular Ca2+ pools had not been depleted. These results corroborate previous results suggesting that Ca2+ is not involved in glycogenolysis stimulation caused by non steroidal anti-inflammatories. PMID- 9226754 TI - Inhibition of rabbit gastric glucosamine synthetase activity by Cu2+, Zn2+ and Se4+. AB - The effects of Fe2+, Cu2+, Zn2+ and Se4+ on the activity of glucosamine synthetase, the rate-limiting enzyme of mucus synthesis, in rabbit gastric corporal mucosa were examined. Cu2+, Zn2+ and Se4+ inhibited the glucosamine synthetase activity at concentrations ranging from 1 to 10 microM (Cu2+, 8-98% inhibition; Zn2+, 10-99% inhibition; Se4+, 32-89% inhibition). The inhibitory effects of these three ions were much stronger than that of UDP-N acetylglucosamine known as a representative inhibitor of the glucosamine synthetase activity (10 microM, 52% inhibition). Fe2+ had no significant effect on the glucosamine synthetase activity up to 100 microM. These results suggest that Cu2+, Zn2+ and Se4+ can be potent inhibitors of gastric glucosamine synthetase activity. PMID- 9226755 TI - Effect of 13-hydroperoxyoctadecadienoic acid on the glucosamine synthetase activity in rabbit gastric mucosa. AB - The effect of 13-hydroperoxyoctadecadienoic acid (13-HPODE) on the activity of glucosamine synthetase, the rate-limiting enzyme of mucus synthesis, in rabbit gastric corporal mucosa was examined. 13-HPODE inhibited the glucosamine synthetase activity at concentrations ranging from 10 to 100 microM. The effect was concentration-dependent, and the concentration required for 50% inhibition was approximately 20 microM. Experiments utilizing Fe2+ and 13 hydroxyoctadecadienoic acid revealed that the inhibitory effect of 13-HPODE on the glucosamine synthetase activity is not due to the alcohol adduct and hydroxyl radical which are expected to be formed from 13-HPODE, and that the hydroperoxyl functional group is a prerequisite. The fact that tert-butyl hydroperoxide exhibited about 50 times weaker inhibition than 13-HPODE indicates the relative specificity of fatty acyl hydroperoxides in the modulation of the glucosamine synthetase activity. These results suggest that 13-HPODE has the potential to modulate the synthesis of gastric mucus by affecting the glucosamine synthetase activity. PMID- 9226756 TI - Metyrapone reductase purified partially from liver microsomes of male rats: the enzyme differs from acetohexamide reductase. AB - An enzyme catalyzing the reduction of metyrapone, a diagnostic drug with a ketone group, was partially purified from liver microsomes of male rats. The partially purified metyrapone reductase had no ability to reduce acetohexamide, an oral antidiabetic drug with a ketone group, even though both metyrapone and acetohexamide are reduced in liver microsomes of male rats. These results clearly indicate that the reduction of these two drugs can be catalyzed by different enzymes. The partially purified metyrapone reductase was found to reduce aldehydes, ketones and menadione. The substrate specificities were in fair agreement with those of carbonyl reductase. However, the partially purified enzyme was strongly inhibited by inhibitors of aldehyde reductase, such as barbital, phenobarbital and sodium valproate. PMID- 9226757 TI - Pharmacokinetics of recombinant human erythropoietin in rabbits and 3/4 nephrectomized rats. AB - The nonlinear pharmacokinetics (1000, 5000, and 10000 IU/kg) and tissue distribution (5000 IU/kg) of erythropoietin (EPO) after intravenous administration of recombinant human EPO (rhuEPO) to rabbits, extent of absolute bioavailability (F) of EPO after subcutaneous administration (5000 IU/kg) to rabbits, and pharmacokinetics of EPO after intravenous administration to 3/4 nephrectomized rats (1000 IU/kg) were investigated. After intravenous administration of rhuEPO, 1000 IU/kg to rabbits, the terminal half-life, t1/2 (296, 368, and 378 min) and mean residence time (255, 318, and 326 min) decreased significantly, however, the total body clearance, CL (0.233, 0.165, and 0.169 ml/min/kg) and nonrenal clearance, CLNR (0.196, 0.141, and 0.120 ml/min/kg) increased significantly when compared with those of 5000 and 10000 IU/kg. The above dose-dependent pharmacokinetic parameters of EPO could be due to saturable metabolism of EPO in rabbits. The affinity of EPO to rabbit tissues studied was very low as reflected to less-than-unity values of tissue to plasma ratios except in the bile. This was supported by a considerably low value of volume of distribution of EPO at steady state (Vss) after intravenous administration of rhuEPO, 1000-10000 IU/kg, to rabbits (0.0524-0.0591 l/kg). After subcutaneous administration of rhuEPO, 5000 IU/kg, to rabbits, the plasma concentration of EPO was reached its peak at 600-720 min and declined slowly with a mean t1/2 of 1040 min. The F value after subcutaneous administration to rabbits was 43.1%. After intravenous administration of rhuEPO, 1000 IU/kg, to control and 3/4 nephrectomized rats, the CL, CLNR, and Vss were not significantly different, however, the MRT and CLR were significantly different between two groups of rats. PMID- 9226758 TI - Effects of chronic cigarette smoke inhalation on the development of senile lung in senescence-accelerated mouse. AB - The senescence-accelerated mouse (SAM) manifests most of the features of function and morphology in the senile lung with aging. However, little is known about the effects of age and cigarette smoke on alterations of the lung in SAM. In the present study, we examined the effects of chronic cigarette smoke inhalation and age on the function and morphology of lungs in two strains of SAM, SAMP2 (senescence-prone strain) and SAMR1 (senescence-resistant strain), from 6 months of age (young) and 18 months of age (aged). After 4 weeks of cigarette smoke inhalation, a small but significant airspace along with a leftward shift of the pressure-volume (P-V) curve was observed in young SAMP2, but not in SAMR1. However, the airspace size of young SAMP2 with cigarette inhalation was smaller than that in aged SAM with air inhalation, suggesting that the effect of age may be greater than that of the small burder of tobacco smoke on the lung alterations in SAMP2. In the aged SAM, there were no differences in function and structure between tobacco-exposed and air-exposed mice. Because the changes in the lungs of young SAMP2 exposed to cigarette smoke were partly simulated with age-related alterations in human lung, and because age-dependent changes of lungs were clearly investigated in SAMP2, this strain may be an interesting animal model for investigating the effects of age and/or cigarette smoke on alterations in lung structure and function. PMID- 9226759 TI - Hepatic adenine nucleotides and microsomal cholesterol 7 alpha-hydroxylase activity in the obstructed and freely draining lobes of the liver after selective bile duct obstruction. AB - BACKGROUND: The effect of selective bile duct obstruction (SBDO) on hepatic reserve function of the bile duct obstructed (BDO) and nonobstructed freely draining (FD) lobes of the liver is obscure. METHODS: The bile duct branches draining from the left lateral and median lobes of the liver were ligated for 4 and 10 days in rats, and hepatic reserve functions in BDO and FD lobes were assessed by microsomal cholesterol 7 alpha-hydroxylase activities and by hepatic adenine nucleotide and energy charge levels. The values were compared with those in sham-operated control liver. Cholesterol 7 alpha-hydroxylase activities were determined by gas-liquid chromatography-mass spectrometry, and hepatic adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) levels with high-pressure liquid chromatography. RESULTS: The histological examination of the BDO lobes showed proliferation and formation of new bile ductules and fibrous connective tissue linking portal areas. Microsomal cholesterol 7 alpha-hydroxylase activities, hepatic energy charge and adenine nucleotide levels did not differ between FD and BDO lobes, and the values were similar to those in the sham-operated liver. CONCLUSIONS: Selective bile duct obstruction shows no adverse effects on microsomal and mitochondrial functions in either BDO or FD lobes of the liver. PMID- 9226760 TI - Local inotropic stimulation by methylene blue does not improve mechanical dysfunction due to myocardial stunning. AB - We tested the hypothesis that reduction of intramyocardial cyclic guanosine monophosphate (GMP) by methylene blue (MB) would improve mechanical dysfunction in stunned myocardium. Regional stunning was produced in nine open-chest anesthetized dogs by a 12-min left anterior descending coronary artery (LAD) occlusion. MB was infused into the LAD during reperfusion (1 mg/kg per min). Stunning reduced LAD force development, introduced a significant time delay between the onset of force and shortening (delay) and caused significant systolic bulging to occur. Stunning reduced systolic regional work (the integrated product of force and segment shortening during systole), but did not significantly alter regional oxygen consumption or cyclic GMP levels. MB decreased cyclic GMP (1.8 +/ 0.2 to 0.9 +/- 0.1 pmol/g) and increased peak force (36 +/- 5 to 55 +/- 10 g). However, MB increased delay (93.9 +/- 18.4 to 233 +/- 19 ms) and systolic bulging (5.9 +/- 2.1% to 9.3 +/- 2.8%) and further reduced systolic regional work (control; 4204 +/- 933 g x mm/min; stunned: 2191 +/- 542 g x mm/min; MB: 1153 +/- 516 g x mm/min). MB increased regional myocardial oxygen consumption (7.4 +/- 1.0 to 15.6 +/- 2.7 ml O2/min per 100 g). These results suggest that depressed contractility, while present in myocardial stunning, is not the primary cause of mechanical dysfunction. PMID- 9226761 TI - Mediators of antigen-induced gastrin release: role of antigen-antibody complexes and the complement system. AB - That orally administered antigen was shown to induce gastrin release in immunized animals was a new aspect of gastrointestinal physiology. The mediators responsible for this immunological effect are still unclear. In an attempt to discover more about the mechanisms regarding antigen-induced gastrin release, we developed an in vitro system where fragments of rat antral mucosa were challenged. This makes it possible to determine the role of antigen-antibody complexes and the complement system in the mechanism of antigen-induced gastrin release. Wistar rats were immunized in vivo with NIP-OVA and mucosal fragments were challenge, in vitro with NIP-HGG. Gastrin was determined after a preincubation and a challenged incubation period without supernatants. After antigenic challenge, supernatants were used for in vitro challenge in order to rule out the presence of a soluble mediator and activation of complement. In a second group of experiments Wistar rats were used to study in vitro the release of specific antibodies after antigenic challenge. With this experimental design we were able to show increased gastrin secretion after antigenic challenge in vitro in the presence of intact tissue. It is shown that the increased gastrin release is most probably mediated by activation of the complement system in the presence of antigen-antibody complexes. These are built up by specific anti-NIP antibodies and NIP-HGG used for the challenge. The complement system might be the final pathway of the observed increased gastrin release. PMID- 9226762 TI - Regulation of male fertility by pyrimethamine in adult mice. AB - Studies were carried out to determine the antifertility and reversibility effect of pyrimethamine (PYR) in adult male mice. The parameters mainly included sperm count and motility, fertility, histoarchitecture of testis and testicular cell kinetics quantitatively following oral administration of PYR (50 mg/kg body weight per day) for 30 days. The same parameters were also studied in PYR-treated animals which were allowed to recover for 45 days (recovery group). The results suggest that sperm motility as well as counts were significantly decreased in PYR treated animals, and the fertility rate fell to zero. Testicular histology as well as germ cell kinetics were altered. However, in the animals of the recovery group, all the parameters studied were more or less similar to those of control animals. The study demonstrates the antifertility as well as reversible efficacy of PYR. PMID- 9226763 TI - Development of T-tube tracts in piglets: effect of insertion method and material of T-tubes. AB - T-tube-related bile leakage is a considerable problem in liver transplantation but rather rare in surgery of biliary lithiasis. To investigate the effect of T tube insertion method and material on the intraperitoneal T-tube tract, we performed a choledochotomy and insertion of T-tube (four of silicone, seven of latex, four of silicone with a latex sheath around the long arm) for 2 weeks on 15 piglets (choledochotomy group), and sutured a transected bile duct over a T tube stent in nine piglets (five silicone, four latex), inserted similarly as in liver transplantations, for 6 weeks. Sixteen patients underwent cholectochotomy and T-tube drainage with a latex T-tube (n = 8) and latex-sheathed silicone T tube (n = 8) for a median 9 (7-21) days. Histological examination of T-tube tracts in piglets was made, and complications after T-tube removal in the latex T tube group were compared with those in the latex-sheathed silicone T-tube group. In piglets, latex T-tubes induced better tracts than silicone T-tubes (P < 0.05). Piglets in the choledochotomy group had tracts superior to those in the anastomotic stent group (P < 0.05). There was one bile leakage in the latex T tube group, and none in the latex-sheathed silicone T-tube group. We conclude that T-tube tract development is affected by both the material and the insertion method of T-tubes. A silicone T-tube with a latex sheath around the long arm may also be a good choice for T-tube material in liver transplantation. PMID- 9226764 TI - Immune responses to mucosal infection: the Helicobacter pylori paradigm. PMID- 9226765 TI - Circulating cell adhesion molecules in HIV1-infected patients as indicator markers for AIDS progression. AB - Cell adhesion molecules play an important role during immune responses. Circulating (c) forms of these molecules have been used as monitors of disease progression. In this study, we have investigated serum levels of ICAM1, ICAM2, ICAM3 and VCAM1 in HIV-infected patients. Our results showed that levels of cICAMs and cVCAM1 are increased during HIV infection. Among an HIV-infected population, the cICAM2 level was higher in the asymptomatic group compared to the AIDS group. In contrast, the cICAM1 level was higher in the AIDS group compared to the asymptomatic group. No difference between the two groups was observed in cICAM3 and cVCAM1 levels. A significant correlation was found between cICAM1, cICAM2 and cVCAM1 in both populations. We also showed that the cICAM1/cICAM2 ratio was correlated with the increase in the c beta 2 microglobulin level and the decrease in CD4 T-cell counts in the AIDS group. These results indicate that serum cCAM1 and cICAM2 in HIV infection could be additional markers to discriminate between asymptomatic and progressor patients. PMID- 9226767 TI - Stimulatory effects of polar glycopeptidolipids of Mycobacterium chelonae on murine haematopoietic stem cells and megakaryocyte progenitors. AB - The effects of polar glycopeptidolipids of Mycobacterium chelonae (pGPL-Mc) on haematopoietic stem cells and on megakaryocyte progenitors in bone marrow (BM) and spleen were investigated in mice. We studied the in vivo spleen colony forming ability and marrow repopulating ability of pGPL-Mc by assays of colony forming units-spleen (CFU-S). The number of CFU-S was increased in BM when both donors and recipients were treated with pGPL-Mc. In contrast, a single treatment of donors induced enhancement of spleen CFU-S. The number of pre-CFU-S was not significantly increased by pGPL-Mc injection. Megakaryocyte (Meg) progenitors were determined in vitro with a quantitative cultural analysis of bone marrow and spleen cells in agar in the presence of spleen-conditioned medium. A statistically significant increase in BM and spleen CFU-Meg was observed two days after the last administration of pGPL-Mc. This experiment points out the ability of pGPL-Mc to induce substantial stimulation of megakaryocytopoiesis and slight proliferation of stem cells in BM, but which is more pronounced in spleen. This molecule therefore appears to be a potential adjuvant of chemo- and radiotherapy in order to palliate the cytotoxic side effects of these cancer therapeutic modalities. PMID- 9226766 TI - Production and metabolism of platelet-activating factor by human bone marrow cells. AB - Platelet-activating factor (PAF) is a phospholipid mediator of inflammation present in the human bone marrow. Freshly isolated human mononuclear bone marrow cells and marrow stromal cell cultures produced PAF under calcium ionophore (2 microM) and LPS (10 micrograms/ml) stimulation. By contrast, M-CSF (1000 U/ml), GM-CSF (100 ng/ml), IL1, IL3, IL6 and stem cell factor (10 ng/ml) did not stimulate PAF production. Marrow stromal cells produced 50-fold more PAF than freshly isolated mononuclear marrow cells, suggesting that stromal cells might be the major source of the human marrow-derived PAF. Mononuclear marrow cells and stromal cell cultures metabolized PAF with 1-alkyl-2-acyl-glycerophosphocholine as the major metabolic product. PMSF and p-BPB decreased the catabolism of PAF by freshly isolated marrow cells, but not by stromal cell cultures. While stromal cells rather than haematopoietic progenitors might be a major source of the human bone-marrow-derived PAF, both cell types metabolize it, suggesting their putative role in the regulation of PAF concentration in the human bone marrow. PMID- 9226768 TI - Malaria at the millenium. PMID- 9226769 TI - Integrating clinical pharmacists into the primary health care team: a framework for rational and cost-effective prescribing. AB - A recent Audit Commission report into general practice prescribing identifies areas where general practitioner and pharmacist collaboration could be beneficial. Two such areas are formulary development and repeat prescribing review. Increased generic prescribing is encouraged in the report and in central priorities for Scottish Health Boards. This study was designed to develop and assess the effects on prescribing, of a practice formulary and a procedure for change to generic name prescribing. A practice formulary, standards for generic name prescribing and an approach to prescribing review were agreed, developed and implemented. Formulary compliance and the extent of prescribing generically and of changes to generic prescriptions were assessed by prospective prescription monitoring. Consultations resulting in a prescription reduced from 69% to 59% and 80% of acute prescribing events were met from 144 formulary medicines. Rapid change to generic name prescriptions was achieved without patient complaints and the overall generic prescribing level increased from 57% to 68%. Eighty percent of all new prescriptions were generic. PMID- 9226770 TI - Quinine--acute self-poisoning and ocular toxicity. AB - Quinine is widely prescribed as a treatment for nocturnal leg cramps. In acute overdosage, it may lead to significant ocular problems. This study reveals that some 30 patients were discharged from Greater Glasgow Health Board hospitals between 1988 and 1992 following acute self-poisoning with quinine. Six of these subjects had severe visual problems, two being registered as blind and another three as partially sighted. These observations, combined with the doubtful clinical efficacy of quinine, suggest that its continued widespread use should be reappraised. PMID- 9226771 TI - Epidemiology and awareness of malaria in the Lothians. AB - Awareness of malaria and its presentation could be improved. Awareness of symptoms, incubation periods and route of transmission was poor, as was compliance with prophylaxis. The mean difference between the time to admission and the time to presentation was 2.9 days which reflects medical rather than patient associated delay. PMID- 9226772 TI - Information about antimalarial chemoprophylaxis in hospitalised patients--is it adequate? AB - Malaria remains a huge public health problem worldwide, with over 100 million new cases annually, causing one to two million deaths. This global problem spills over into the UK, with around 2000 cases of reported annually. The proportion of infections due to Plasmodium falciparum (PF) continues to increase and worse still accounts for five to 12 deaths per year. In 1992, Nathwani et al reported the 10 year experience of malaria cases admitted to the Regional Infection Unit, in Aberdeen, Scotland--the "Oil Capital". This study was of interest in that 46% of those British residents who acquired infection had travelled to West or Central Africa on oil related business. The Oil boom of the 1980's appeared to very much centred around Aberdeen and the neighbouring hinterland but did not appear to extend to Dundee which was only 60 miles further down the North-East coast. We, therefore, carried out a retrospective study of patients with malaria admitted to the Regional Infectious Diseases Unit in Dundee over a fifteen year period between 1980 and 1994. PMID- 9226773 TI - Persistent nephrogenic diabetes insipidus following lithium therapy. AB - We report the case of a patient who developed severe hypernatraemic dehydration following a head injury. Ten years previously he had been diagnosed to have lithium-induced nephrogenic diabetes insipidus, and lithium therapy had been discontinued. He remained thirsty and polyuric despite cessation of lithium and investigations on admission showed him to have normal osmoregulated thirst and vasopressin secretion, with clear evidence of nephrogenic diabetes insipidus. Lithium induced nephrogenic diabetes insipidus is considered to be reversible on cessation of therapy but polyuria persisted in this patient for ten years after lithium was stopped. We discuss the possible renal mechanisms and the implications for management of patients with lithium-induced nephrogenic diabetes insipidus. PMID- 9226774 TI - Listeria monocytogenes: a rare cause of pleural effusion in a patient with congestive cardiac failure. AB - We report the case of a 65 year old immunocompetent man with a listerial pleural effusion. Infection of the pulmonary parenchyma and pleura with listeria monocytogenes has been reported in small numbers of immunocompromised patients but there have been only two previous reports of pulmonary listeria in non compromised hosts. PMID- 9226775 TI - A repeat audit of hospital discharge letters in patients admitted with acute asthma. AB - Subsequent to the implementation of recommendations from a previous audit of acute asthma admission discharge letters from our specialist respiratory unit, a repeat audit of typed discharge letters of 86 patients (33 male, mean age 29 SD 9 years) admitted with acute asthma to the same unit over a 12 month period was performed. There was significant improvement in the discharge letter documentation of precipitating factors (p < 0.001), previous admissions with acute asthma (p < 0.01), admission arterial blood gas analysis (p < 0.001), admission peak flow rates (p < 0.05), discharge peak flow rates (p < 0.001), corticosteroid (p < 0.01) and inhaled beta 2 agonist (p < 0.01) prescription on discharge and on the specification of inhaler delivery device on discharge (p < 0.001). No significant differences in discharge letters were found in the documentation of acute therapy or post discharge follow up plan. The improvement in discharge letter quality was attributed to closing the feed back loop from the previous audit though continuing deficiencies in discharge letter contents have been identified again. These deficiencies need to be rectified and the results reaudited. PMID- 9226776 TI - The impact of cost-effectiveness on public and private policies in health care: an international perspective. Introduction and overview. PMID- 9226777 TI - Managed care pharmacy, socioeconomic assessments and drug adoption decisions. AB - A telephone survey of a representative national sample of 51 large managed care organizations in the U.S. (> 50,000 enrollees) was undertaken (1) to understand the role of socioeconomic assessments on drug adoption decisions; (2) to determine the sources of these assessments and the reliance of managed care pharmacy on each; and (3) to determine the resources for internally versus externally performed drug assessments. Socioeconomic assessments (clinical effectiveness, safety, cost of treatment, cost-effectiveness, and quality of life) are often tied to formulary decisions. Plans differ in their use of externally available socioeconomic assessments and in their ratings of the importance to decision making of drug assessments from the various sources. Those using a specific source of drug assessment information rated them in the following order of importance: PBM assessments, other HMOs, peer reviewed literature, evaluations performed by industry, articles in non-peer reviewed publications and, lastly, government reports. Timeliness and comprehensiveness are important components of the overall utility of information. A high percentage of plans reported using some of the various types of assessments, with clinical effectiveness most common, and cost-effectiveness second. The percentage of new drugs that undergo assessments in each of the plans covers a broad range, with 57% of the plans evaluating at least half of all new drugs. All but one surveyed managed care plan reported having either implemented or plans to implement a disease management program. Eighty percent of those surveyed are more concerned about drug assessments than in the past and 88% anticipate greater future use. Although 38 plans (75%) have a person in the organization responsible for drug assessments, this is the primary job in only 14 plans (37%). With greater reliance on drug assessments in the future, there are substantial opportunities for integrating drug assessments, formularies and disease management programs. PMID- 9226779 TI - Pharmacy benefit management, cost-effectiveness analysis and drug formulary decisions. AB - Pharmacy benefit management companies (PBMs) have evolved over the past decade in response to the increased demand for health care cost containment. Their activities include the implementation of drug formularies and the negotiation of rebates from manufacturers. Our analysis of this industry is based on interviews and materials provided by the top five ranked PBM companies which account for over 80% of beneficiaries covered within formulary plans. The formularies of these companies are relatively inclusive, but they are becoming more restrictive over time. At present the use of cost-effectiveness (C-E) studies in the formulary decisions of PBMs has been limited. In this regard, the surveyed PBMs emphasized that most C-E studies have not compared therapeutic substitutes in populations with characteristics that are similar to those of their clients. Pharmacy benefit management companies also have had strong incentives to focus narrowly on drug costs because they typically manage drug benefits on a "carved out" basis. However, PBMs anticipate a growing future role in the integrated management of patient care (disease management) for certain high cost chronic diseases and conditions. All of the leading firms we surveyed have disease management programs in development. The importance of C-E studies to PBM decisions is expected to increase significantly as disease management programs are implemented. The data infrastructure inherent to the PBM industry and the increasing number of employees with advanced training in pharmacoeconomics will permit firms to perform their own internal C-E studies. They are also establishing various alliances and joint ventures with drug manufacturers, health maintenance organizations, and academic institutions to perform these analyses. The leading PBMs tend to favor active participation in the development of methodological approaches to C-E studies over government regulations such as those proposed by the FDA in 1995. PMID- 9226778 TI - Hospital pharmacy decisions, cost containment, and the use of cost-effectiveness analysis. AB - The key hypothesis of the study was that hospital pharmacies under the pressure of managed care would be more likely to adopt process innovations to assure less costly and more cost-effective provision of care. We conducted a survey of 103 hospitals and analyzed secondary data on cost and staffing. Compared to the size of the reduction in length of stay, changes in the way that a day of care is delivered appear to be minor, even in areas with substantial managed care share. The vast majority of hospitals surveyed had implemented some form of therapeutic interchange and generic substitution. Most hospitals used some drug utilization guidelines, but as of mid 1995 these were not yet important management tools for hospital pharmacies. To our knowledge, ours was the first survey to investigate the link between hospital formularies and use of cost-effectiveness analysis. At most cost-effectiveness was a minor tool in pharmaceutical decision making in hospitals at present. We could determine no differences in use of such analyses by managed care market share in the hospital's market share. One impediment to the use of cost-effectiveness studies was the lack of timeliness of studies. Other stated reasons for not using cost-effectiveness analysis more often were: lack of information on hospitalized patients and hence on the potential cost offsets accruing to the hospital: lack of independent sponsorship, and inadequate expertise in economic evaluation. PMID- 9226780 TI - The Oregon experiment: the role of cost-benefit analysis in the allocation of Medicaid funds. AB - The state of Oregon decided to cover all potentially eligible Medicaid citizens to 100% of poverty. Previously, Oregon had covered persons up to 67% of poverty. In order to keep overall program costs in check. Oregon decided to limit the number of services that its Medicaid program would cover. Oregon's normative choice was to contain program costs by covering all eligible persons up to 100% of poverty, while at the same time uniformly limiting access to certain services for everyone in the overall group of eligible persons. The state developed a prioritization list of medical services and priced the components on the list. The amount of money ultimately available for the Medicaid program was a political decision informed by data about the cost of different services and influenced by the priorities set through an independent process of priority-setting. Physicians were asked to determine what works medically, how well it works, and what benefits accrue to patients. Recognizing that physician perspectives on efficacy might vary from patients' perspectives on valuation of benefits, Oregon's planners developed a method for valuing medical outcomes that stemmed from particular medical interventions. This blend of medical fact and value to patients allowed for comparing valuations by introducing cost considerations. Condition-treatment (CT) pairs linked a medical condition with one or more courses of treatment. The goal was to determine the likely incremental medical benefit from a given treatment. In addition, Oregon developed a Quality-of-Well Being scale to determine the net patient benefit from medical intervention and used a telephone survey to value that net benefit. A cost-benefit ratio was derived, and a prioritization of CT pairs was developed. The article analyzes and evaluates Oregon's use of cost-benefit calculations in the allocation of Medicaid funds, noting that Oregon itself backed away from many of the implications of its cost-benefit analysis and that the Americans with Disabilities Act has constrained use of quality-of-life judgments in Medicaid resource allocation decision-making. PMID- 9226781 TI - Making economic evaluations respectable. AB - Policy-makers worldwide are on a quest to control national spending for health care and to enhance the value received for whatever is being spent on health care. One should think that the economic evaluation of clinical practice would play a major role in this quest. Alas, so far it has not, in spite of considerable progress in the development of suitable methodology for such evaluations. The central point of this paper is that the sheer conceptual and practical complexities of economic evaluations in this context are not the only and possibly not the major barrier to a more widespread use of this type of analysis. Just as important may be the suspicion among lay persons that such analyses are easily driven by the assumptions the analyst packages into the analysis which, in turn, opens economic evaluation to hidden bias toward favored results. It is proposed in this paper that this particular barrier to the use of economic evaluations in health policy could be overcome if these analyses were more routinely subjected to the rigorous and penetrating audits that are customary in financial accounting. Typically, research papers in economics are audited through peer review only as to the methodology employed. The suggestions here is that a proper, respectable audit ought to penetrate all the way to the data that were used to produce the findings in a study. The paper concludes with some suggestions on how to develop such an audit infrastructure. PMID- 9226782 TI - Australian economic evaluation and government decisions on pharmaceuticals, compared to assessment of other health technologies. AB - In this paper the first theme is the experience with the routine use of cost effectiveness analysis in decisions by the Australian Pharmaceutical Benefits Advisory Committee on whether drug products should attract a government subsidy. As a second theme, the contrasting experience with several other health technologies is presented, with economic analysis being less frequently used in a system where there is a weaker regulatory framework. Some general points that emerge in both areas are the importance of factors other than economic evaluation in the decision-making process, and the need to make policy and administrative decisions on the basis of limited data. There is limited material available in the public domain on the interaction of economic evaluation and Australian policy on health technologies. It has been necessary, particularly in relation to the case studies presented here, to rely on input from discussion with a number of individuals and on observations made during personal involvement with some of the assessments. It is not possible to offer substantive evidence in support of this material, and indeed firm evidence in the area of impact of assessments on health policy remains difficult to collect. PMID- 9226783 TI - Economic evaluation under managed competition: evidence from the U.K. AB - Although economic evaluation in health care has a long-standing tradition in the United Kingdom, very little is known about its impact on decision making, particularly following the introduction of the internal market. Since managed competition appears to be growing in popularity worldwide, the U.K. is an interesting case study, as the reforms are well underway and there have been a number of efforts to conduct and disseminate economic evaluations. In this paper the potential for using economic evaluation in health care decision making in the U.K. is discussed. Then its actual impact is assessed in two ways. First, two case studies are discussed, on heart transplantation and the use of pharmaceuticals in the management of labour in pregnancy. Second, new data from a recent survey of potential users of economic evaluations are presented, with the emphasis on exploring the reasons for the impact, or lack of impact, of economic results. It is concluded that the NHS reforms increase the potential for the use of economic evaluation. However, there is a need to increase decision makers' awareness of economic studies and to help them interpret study methodology and results. Although worries about validity of economic studies are one of the major barriers to their use, other important barriers relate to the multiple objectives being pursued, of which increased efficiency is just one, and the difficulties of freeing resources from existing services in order to divert them to more cost effective treatments and programmes. PMID- 9226784 TI - Economic evaluation of medical technologies in Sweden. AB - This paper reviews the use of economic evaluations in the Swedish health care system. The most important actors are defined and examples are given how economic evaluations have played a role in the decision making process. The introduction of extracorporal shock-wave lithotripsy (ESWL) is used as an example on how economic evaluation was used for recommendations to the county councils to adopt this technology. Mammography is used as an example of how the evaluation is used in the political process following an initiative in the parliament. A study of the cost-effectiveness of hypertension treatment illustrates how economic evaluations are included as part of a medical technology assessment by SBU, the Swedish Council on Technology Assessment in Health Care. The role of economic evaluations for drug reimbursement and pricing is also reviewed. The main conclusions are that economic evaluations are one of several factors influencing a decision making process that have a strong strive for consensus. It is thus difficult to make a definitive statement of the contribution of such study to the outcome of the decision making process, and there is no evidence that the evaluation was the decisive factor. However, a number of changes in the way resources are allocated in the Swedish health care system speaks for an increasing role for such studies in the future. The county councils are identified as the main target for economic evaluations, and SBU has a key role in supplying the county councils with high quality assessment of new and old technologies. PMID- 9226785 TI - Economic evaluation in support of national health policy: the case of The Netherlands. AB - This article focuses on economic evaluation as an instrument to support national health policy in the Netherlands. National health policy concerns decision-making on which technologies may enter the health care market, which health care services are included in the package of health care benefits and under what conditions, and what the geographical distribution of medical services and facilities should be. Regarding the latter two issues in particular, the actual and potential role of economic evaluation in health policy is discussed. From the Dutch experience we can learn that a close cooperation between researchers and policy-makers helps to enlarge the impact of economic appraisal on policy-making and that incorporation of the right (financial) incentives is vital to the use of economic appraisal in health care decision-making. PMID- 9226786 TI - Economic evaluation of medical technologies: from theory to practice--the German perspective. AB - This paper briefly describes the German health care system and the role of economic studies in medical technologies and drugs in a decentralized health care system like Germany's. It also provides a number of examples of studies which have been conducted recently in Germany and shows the variation of sponsors, study types, study perspectives and objectives of those studies. It concludes with an outlook on the future role of pharmaco-economic studies in Germany and the recently published German recommendations for conducting those studies. PMID- 9226787 TI - Pharmaceutical economy and the economic assessment of drugs in France. AB - An overview of the present state of development of pharmacoeconomics in France is given, and more specifically how it works, in what context it operates, what kind of studies it produces, what kind of results it reaches, and what kind of future it may expect. The main features of the French health care system and French drug economy are described. Six concrete case studies of pharmacoeconomic assessment are then summarised. The economic problem raised by these new well-known drugs, the characteristics and the conclusions of the studies that have been undertaken and the decisions that have been made by health public authorities are presented. Finally presented are some views about the future of what we term the "institutional pharmacoeconomics", which is a new discipline used by pharmaceutical companies to demonstrate to institutional buyers the value for money of their new products, and to regulate prices. PMID- 9226788 TI - Space, sterility and surgery: circuits of hygiene in the operating theatre. AB - The spaces of the surgical operating theatre (ST) and associated built environment are analysed, to explore what the physical layout means for the interactions which take place. Three "circuits of hygiene" of surgical staff, surgical instruments, and patients are documented, and analysis of these physical movements through the surgical spaces examined for their contribution to sterility. It is concluded that the built environment of the ST contributes reminders to staff to fulfil the necessary procedures of aseptic technique, to ensure the safe passage of the patient through surgery into a condition where s/he may be designated as "healed". PMID- 9226789 TI - Household cost of seeking malaria care. A retrospective study of two districts in Ghana. AB - Although malaria or fever (as it is commonly referred to) is a major cause of morbidity and mortality in Ghana, the cost of treating the disease in the country has not been well documented. Knowledge about the cost of treating malaria can affect the health care seeking behaviour of people and justify increased expenditure for malaria control. This study used data collected from 1289 households in two districts in Ghana to estimate the direct and indirect costs of malaria treatment. Malaria was ascertained not by parasitological tests but through symptoms described by the respondents using a recall period of one month. It was found that substantial amount of time was spent in seeking malaria care and taking care of the sick, which makes the indirect cost per case of fever represent 79% of the total cost of seeking treatment in the survey area. The results provide ample economic justification for malaria control. The average cost of treating an episode of the disease including direct costs and the opportunity costs of travel and waiting time amounted to $8.67 or 3.7 days of male output or 4.7 days of female output. When compared with the average five days loss of output for the patient due to malaria morbidity and caretaking, it can be concluded that the cost of controlling malaria is lower than lost earnings or the value of output. PMID- 9226790 TI - A qualitative study of sexual harassment of female doctors by patients. AB - This paper reports the qualitative data from a study of sexual harassment of female family physicians by patients. In addition to the everyday harassment that any woman might encounter in a work setting, the physicians in this study also reported types of harassment which are unique to the practice of medicine. These include opportunistic harassment such as exposure of the genitals, inappropriately touching the physician when the examination requires close contact, excessive discussion of sexual matters for apparent erotic gratification, and acting out behaviours from non-competent patients. Other reported behaviours were not, strictly speaking, sexual harassment but were troublesome nonetheless, including spontaneous erections during physical examinations, physically intimidating behaviour, and ambiguous behaviours which were sexual in nature, but difficult to interpret. The findings are discussed in the context of theory pertaining to contrapower harassment. It is concluded that for some patients the gender of the physician takes precedence over her occupational status and, this combined with the unique characteristics of the doctor/patient relationship, can make the practice of family medicine more conductive to sexual harassment than other professions. PMID- 9226791 TI - Social network associations with contraceptive use among Cameroonian women in voluntary associations. AB - This paper examines the association between social networks and contraceptive use. Using data from a survey of women belonging to voluntary associations in Yaounde, Cameroon, we find that the behavior and characteristics of the members of a respondent's personal networks are associated with her contraceptive use, over and above a set of her own individual characteristics that are usually found to be important. Respondents who report that their network partners approve of contraception, use it, and encourage the respondent to use are more likely to use contraception themselves; the association with encouragement is particularly strong. Moreover, there is a strong association between the specific methods of contraception used by a respondent and those used by her network partners, suggesting that members of personal networks exchange and evaluate specific methods. Because most of the respondent's network partners were interviewed, we are able to compare the respondent's perceptions of contraceptive use by her network partners with the network partner's actual use. We find that it is perceptions of use that matter, even if those perception are incorrect. PMID- 9226792 TI - The conceptual basis of ethnic group terminology and classifications. AB - "Ethnic group" is a problematic variable in health-related research. While self identification is now widely accepted as the appropriate mode of assignment, the impracticalities of a free response in the collection of ethnic group data mean that categorisation into a limited set of choices must take place. The substantial and increasing number of persons in minority ethnic groups who identify through non-standard responses emphasises the need to develop classifications that accommodate salient vernacular terminology. The use of informants in cognitive settings and the monitoring of open-ended responses appear to offer the best way of determining which group labels to employ. The recommended approach addresses the research priorities for accurate, consistent, and high quality data and is also responsive to the dynamic nature of ethnic group and the growing ethnic diversity of the population. PMID- 9226793 TI - Patients' and professionals' understandings of the causes of chronic pain: blame, responsibility and identity protection. AB - A social constructionist analysis of how sense is made of the causes of chronic pain is reported. It is recognised that there is a multiplicity of stories available in any culture from which understanding can be reached. Q-factor analysis is used within a critical framework as Q-methodology. Sixty chronic pain patients and pain professionals completed the sorting procedure. Four factors were derived that account for the causes of chronic pain. These are reported as the patients' account, the professionals' account, the scientists account and the alternative practitioner's account. Common to all four accounts are the themes of responsibility, blame and the need to protect identity. It is argued that in all accounts responsibility is repositioned away from the sufferer or the healer. In all of the accounts blame is resisted or deflected away from individual ownership. Finally, it is argued that when pain is no longer useful as a symptom, identity is challenged, weakened and at risk for both chronic pain patients and pain professionals. Implications of this study for chronic pain research and treatment are discussed. PMID- 9226794 TI - Gender differences in medical treatment: the case of physician-prescribed activity restrictions. AB - A growing scientific literature highlights concern about the influence of social bias in medical care. Differential treatment of male and female patients has been among the documented concerns. Yet, little is known about the extent to which differential treatment of male and female patients reflects the influence of social bias or of more acceptable factors, such as different patient preferences or different anticipated outcomes of care. This paper attempts to ascertain the underlying basis for an observed differential in physicians' tendency to advice activity restrictions for male and female patients. We explore the extent to which the gender-based treatment differential is attributable to: (1) patients' health profile, (2) patients' role responsibilities, (3) patients' illness behaviors, and (4) physician characteristics. These four categories of variables correspond to four prominent social science hypotheses concerning gender differences in health and health care utilization (i.e, biological basis hypothesis, fixed role hypothesis, socialization hypothesis, physician bias hypothesis). Data are drawn from the Medical Outcomes Study (MOS), a longitudinal observational study of 1546 patients of 349 physicians practicing in three U.S. cities. Multivariate logistic regression is used to evaluate the likelihood of physician-prescribed activity restrictions for male and female patients, and to explore the absolute and relative influence of patient and physician factors on the observed treatment differential. Results reveal that the odds of prescribed activity restrictions are 3.6 times higher for female patients than for males with equivalent characteristics. The observed differential is not explained by differences in male and female patients' health or role responsibilities. Gender differences in illness behavior and physician gender biases both appear to contribute to the observed differential. Female patients exhibit more illness behavior than males, and these behaviors increase physicians' tendency to prescribe activity restrictions. After accounting for illness behavior differences and all other factors, the odds of prescribed activity restrictions among female patients of male physicians is four times that of equivalent male patients of those physicians. Medical practice, education, and research must strive to identify and remove the likely unconscious role of social bias in medical decision making. PMID- 9226795 TI - Salary inequality and primary care integration in South Africa. AB - Separation of curative and preventive health programmes often impairs the coordination of primary care in developing countries. Salary differentials between organisations may aggravate non-cooperation. Implementation of a unitary national health service by South Africa's first democratically elected government has been hampered by salary differences, but no organisation possessed information on their magnitude. This paper reports on a study which estimated the distribution and conditions of service of all 224,000 public health sector personnel in South Africa, modelled options for equalising salaries between health authorities, and considered the financial and political feasibility of the options. The most important salary differential was between provincial and local authority nurses. The option to increase salaries selectively for personnel in rural and primary care would be most feasible and most in keeping with government plans. Health service unions face conflicts of interest, and professional organisations may oppose changes in nurses' roles. In a rapidly changing health system with fragmented managerial information, a combination of administrative survey, quantitative modelling and policy analysis helped clarify a key obstacle to reform. The South African case is a warning to other countries that decentralised pay bargaining may result in uncoordinated care which may be costly and difficult to overcome. PMID- 9226796 TI - Quality of sleep during economic recession in Finland: a longitudinal cohort study. AB - To assess the association between the economic recession of the 1990s in Finland and sleep behaviour, a longitudinal study was conducted in an adult Finnish population cohort. Baseline data were obtained by means of reports on sleep behaviour, health-related behaviour, health status, and objective laboratory tests in 1983-1987. The second screening conducted in 1992-1995, i.e. during economic recession, repeated data collection by postal questionnaires. The prevalences of various sleep symptoms including insomnia, daytime tiredness, fatigue, parasomnias and the use of hypnotics remained similar in the same age cohorts during economic recession. Alcohol consumption and snoring increased among the middle-aged (30-49 years), though snoring shows the greatest individual stability among various sleep symptoms. Despite some baseline differences in the sleep/health behaviour frequencies, the changes were independent of gender and socioeconomic class. The prevalences over eight years of insomnia and snoring show fair chronicity, whereas daytime tiredness and fatigue seem to be less chronic. Middle-aged participants who were stably employed at the initial screening but became unemployed during economic recession were studied separately. Prospectively unemployed persons suffered more from insomnia and used more hypnotics than the continuously employed. We conclude that the sleep quality of the general Finnish population has not drastically deteriorated during severe economic recession except among unemployed blue-collar workers. PMID- 9226797 TI - Explaining disablement in modern times: hand-injured workers' accounts of their injuries in Hong Kong. AB - In this paper, an attempt is made to explore how people respond to injuries at work among a group of hand-injured workers in the context of contemporary Hong Kong society. Qualitative research methods were adopted. A "focused interview" by means of open-ended questions was employed to investigate those workers who have suffered from work-related injury for over 10 years. Concerning the perceived cause of the injury, the industrial production process such as machine defects, piecework, limited working experience, and lack of supervision triggered the onset of the injuries which resulted in permanent disability. At the same time, respondents tellingly explained their injury in terms of magical-religious forces such as fate and luck. This kind of understanding is embraced in cultural beliefs that are commonly found among Chinese. Such understandings appear fatalistic but allow the individual to actively cope with such misfortune. The emphasis on harmony and stability among Chinese also affected what action they took against the employer for negligence. It was shown that in general many respondents were bound by kuan-hsi (personal relationship) and tended to preserve the harmony between themselves and their employer. In modern societies like Hong Kong, traditional values and modern practices are complementary to each other in coping with life stresses such as disablement. Modernity does not provide a solid means for people to have more control over their lives. Many people still stick to their habituated lifestyle as a guiding light in day-to-day matters. Concerning risk assessment in relation to work safety, workers' value systems and practices on the shopfloor should be taken into consideration. PMID- 9226798 TI - Climacteric hormone therapy in medical and lay texts in Finland from 1955 to 1992. AB - The "social shaping of technologies" approach holds that a technology is both socially embedded and that it shapes the social environment. The aim of this study was to investigate how hormone therapy use during the climacterium and subsequently was socially shaped in texts published in the main Finnish medical journals and lay magazines during 1955-1992. In these two arenas physicians, especially gynecologists, played the major role in the debate and their professional knowledge on hormone therapy was mixed with their views on women's status and roles, the quality of life and fears about aging when they were promoting hormone use, especially in the lay magazines. This type of argument for the promotion of hormone use persisted in the most recent texts, despite the availability of substantial evidence both for the against of hormone therapy. Overall, the texts clearly favored the benefits of the therapy. Three periods of differing orientation can be discerned. Attitudes towards hormone therapy tended to be cautious from 1955 through the 1970s, more enthusiastic in the 1980s, and mixed at the start of the 1990s. In the most recent texts critical comments came from individual women who had used the therapy or decided not to, including female physicians and other professionals. The results suggest that hormone therapy is socially embedded, but may also shape perceptions and the understanding of women's aging. The social shaping of the technology approach may improve our understanding of the development of health policy towards women at and after the age of the climacterium. PMID- 9226799 TI - Comparing measures of health inequality. AB - Several methods are available to measure social inequalities in health. This paper discusses the advantages and disadvantages of different approaches, in particular the odds ratio, the slope and alpha. These methods are illustrated using data from subjects in the 1958 British birth cohort. The inequality measures are compared using health status at ages 23 and 33. Six health indicators are examined, including self-rated health, limiting long-standing illness, psychological health, respiratory symptoms, asthma and obesity. Two social indicators are compared, namely class at birth and educational qualifications. Conclusions do not differ substantially using the three methods for measuring inequality. However, consistent differences were evident between the measures of social position, with greater inequalities apparent for educational qualifications. Choice of social indicator therefore appears to be of primary importance in measuring health inequality. PMID- 9226800 TI - Social support as conversation: analysing breast cancer patients' interactions with their partners. AB - This paper demonstrates an alternative approach to studying the process of social support in close relationships by examining three tape-recorded conversations between breast cancer patients and their partners. We argue that existing research fails to take account of the "social" aspects of social support, and that new methods, sensitive to the complexities of human interaction, are needed. One promising method, tape-assisted recall (in which an audiotape of an interaction is played back to participants), allows researchers to examine the help-intended communication within a dyad as well as each participant's view of that communication. The conversations of the three couples and the participants' moment-by-moment perceptions of these conversations illustrate the complexities of help-intended communication. The commentaries in the tape-assisted recall sessions yielded insights into the interactions that were not always apparent on the surface: the personal meanings of the interactions, in the context of the couple's relationship, were necessary for understanding how support attempts are delivered effectively and why they sometimes fail. The strengths and limitations of the method for the study of social support are discussed. Implications for community health interventions to optimize support for couples coping with illness are also addressed. PMID- 9226801 TI - Psychosocial study of epilepsy in Africa. AB - As documented by many authors, the social position of epileptics in many small scale societies of Africa is marginal at best, and is often characterized by rejection, discrimination, even ostracism. Such negative and noxious attitudes toward persons suffering from epilepsy are rooted in traditional beliefs about causes and nature of convulsive disorders and these have parallels in European history. This article focuses on the psychosociocultural aspects and indigenous concepts of epilepsy, on popular attitudes towards, and social status of, sufferers from epilepsy in a Tanzanian tribal population. The authors present a comparative analysis of focus group discussions conducted with epileptics and with matched controls in two isolated communities. In one community (Mahenge) a clinic for epilepsy has been operating for over 36 years, with a public education component during the last four years, whereas in the other community (Ruaha) epileptics have only been sporadically treated in a small mission dispensary and people have had little opportunity to learn about the nature and modern treatment of convulsive disorders. The responses obtained in focus group discussions reflect the significant change in notions about the illness, in the attitude toward and in the social status of epileptics in Mahenge, while the people of Ruaha still regard epilepsy as a typical "African" affliction fraught with supernatural danger and not effectively treatable by modern medicine. PMID- 9226802 TI - Social support in pregnancy can reduce the incidence of low birthweight deliveries. PMID- 9226803 TI - Outline of A. V. Palladin Institute work. PMID- 9226804 TI - Inhibitory effect of D and DD fragments on fibrin polymerization. PMID- 9226805 TI - C-terminal effect of B beta-chain of fibrinogen on fibrin polymerization. PMID- 9226806 TI - Purification and characterization of ecamulin--a prothrombin activator from the venom of multi-scaled viper (Echis multisquamatus). PMID- 9226807 TI - The mechanism of fibrin polymerization sites functioning. PMID- 9226808 TI - Investigation of the plasminogen-streptokinase interaction. PMID- 9226809 TI - Parameters of haemostasis in the elderly people. PMID- 9226810 TI - Quasi-elastic light scattering spectroscopy for investigation of plasma proteins. PMID- 9226811 TI - Haemostasis studies related to gastric abdominal haemorrhages. PMID- 9226812 TI - Formation and dissolution of fibrin in model systems. PMID- 9226813 TI - Computer modelling of fibrin assembly. PMID- 9226815 TI - Fibrinolysis and its clinical importance. PMID- 9226814 TI - The influence of 6-AHA on the fibrin clot degradation initiated by streptokinase in normal and alpha 2-antiplasmin-deficient plasma. PMID- 9226817 TI - Fibrin clot architecture. PMID- 9226816 TI - An enzyme immunoassay for polymorphonuclear leukocyte-mediated fibrinolysis. PMID- 9226818 TI - Biochemistry and measurement of fibrinogen. PMID- 9226819 TI - Plasminogen variants and streptokinase binding. PMID- 9226820 TI - Thrombolytic therapy (a review). PMID- 9226821 TI - Therapeutic use of fibrin--a new class of fibrin sealant with minimal risks. PMID- 9226822 TI - Therapeutic use of streptokinase. PMID- 9226824 TI - Independently folded domains in fibrinogen. PMID- 9226823 TI - The effect of fibrinogen polymerization and crosslinking on exposure of fibrin specific epitopes and t-PA mediated plasminogen activation. PMID- 9226825 TI - Domain structure and interactions of plasminogen activators. PMID- 9226826 TI - Plasminogen conformations. PMID- 9226827 TI - Soluble fibrin. Characterisation and assay procedures. PMID- 9226828 TI - Fibrin polymerisation. Evidence for a secondary polymerisation site on the carboxy terminal end of the AA chain using a human fibrin specific murine monoclonal antibody. PMID- 9226830 TI - On the role of effector site in serine proteinases hydrophobic chromatography. AB - Causes of differences between the calculated and experimentally obtained properties of affine sorbents with low-molecular ligands. The effectory character of the so-called hydrophobic chromatography of serine proteins and the role of a spacer in this process are supposed. PMID- 9226829 TI - Character of structural disturbances caused by chloroalkylamines in the model and erythrocyte membranes. AB - beta, beta-Dichlorodiethylamine is proved not to induce structural disturbances in phosphatidylcholine liposomes and erythrocyte membranes which is registered by fluorescence methods. Methyl-beta, beta-dichlorodiethylamine and metaxylyl-beta, beta-dichlorodiethylamine cause the increase in microviscosity of lipid bilayer hydrophobic areas in both erythrocyte membranes and liposomes. Besides, polarity of the latter also decreases, and the metaxylyl derivative alkylates nucleophilic centers of phospholipid phosphate groups in liposomes. Erythrocyte membranes, being treated by beta, beta-dichlorodiethylamine derivatives, the increase in the membrane protein hydrophobicity is registered as well as the decrease in their immersion in the lipid bilayer. PMID- 9226831 TI - Epidemiology of Rhodococcus equi infections: a review. AB - An overview of epidemiology of R. equi infection in foals is presented, emphasizing the importance of the virulence-associated antigens and plasmids as epidemiological markers. The monoclonal antibody-based colony blot test has been developed to identify rapidly and accurately virulent R. equi. Epidemiological studies conducted during the recent 5 years have revealed that: (1) avirulent R. equi are widespread in the feces of horses and their environment on every farm; (2) the feces of horses and the environment of the horse farms having endemic R. equi infections demonstrated heavy contamination with virulent R. equi, but the farms without the problem did not, thus suggesting that foals bred on a farm with endemic disease are exposed more frequently to virulent R. equi in their environment than those of a farm without the problem; (3) only virulent R. equi are isolated from lesions of naturally infected foals, showing that natural infections in foals are principally by virulent R. equi, but not avirulent organisms; (4) infected foals which constantly shed large quantities of virulent R. equi in their feces are the major source of virulent R. equi, which this may be the mechanism of progressive development of infection on farms with a history of the disease. At present, farms with a potential for endemic infection can be distinguished on the basis of the contamination with virulent R. equi, so regular examination of foals and their environment by virulence markers might be the most practical approach to control R. equi infection on endemic farms. PMID- 9226832 TI - Immunity to Rhodococcus equi. AB - Rhodococcal pneumonia is an important, life threatening disease of foals and immunosuppressed humans. Increased knowledge of the mechanisms of protective immunity are required in order to develop an effective immunoprophylaxis strategy for horses and immunotherapeutic regiments for people. Both humoral and cellular components of the immune system may be involved in immune clearance of R. equi. The susceptibility of foals less than 4-6 months of age is postulated to reflect waning maternal antibody, and passive transfer of hyperimmune plasma can provide protection on endemic farms. However, effective clearance is likely to require appropriate cellular responses, including the secretion of cytokines. In murine models, both CD4+ and CD8+ T lymphocytes can reduce bacterial counts in the lung. CD4+ cells appear to be both required and sufficient, and IFN-gamma is a primary mediator. Clearance appears to be a type 1 immune response while type 2 responses may lead to a failure to clear and lesion development. It remains to be determined how the cellular immunity experiments reported in mice relate to horses and humans. Likewise, the role of specific R. equi antigens in protective immunity has not been determined. PMID- 9226833 TI - Protective effect against Rhodococcus equi infection in mice of IgG purified from horses vaccinated with virulence associated protein (VapA)-enriched antigens. AB - IgG was purified from horses immunized with repeated doses of virulence associated (VapA) enriched antigens extracted with Triton X-114 from the surface of a virulent strain of R. equi. This IgG were administered to mice immunosuppressed by prior treatment with indomethacin. Mice administered the higher dose were completely protected against intraperitoneal infection with R. equi; mice given the lower dose were partially protected. By contrast, mice administered concentrated nonimmune equine IgG were not protected. This study demonstrates that VapA may be an important antigen involved in humoral protective immunity in R. equi infections caused by foal virulent strains. PMID- 9226834 TI - Immunoprophylaxis of Rhodococcus equi pneumonia in foals. AB - An immunoprophylaxis program for R. equi infection of foals has been established on a number of thoroughbred breeding farms in Argentina over the past 4 years. Nearly 800 mares annually were immunized subcutaneously during the last 2 months of pregnancy with 2-3 doses of a vaccine containing soluble antigens of R. equi, including the virulence associated protein (VapA) and 'equi factors' exoenzymes. The mortality from R. equi pneumonia in the foals from vaccinated dams dropped from an average of 3% in the 5 years before the vaccination program was initiated to an average of 1.2% in the 4 years during which the program was applied (P < 0.02). On 3 farms, an additional 380 foals of vaccinated dams annually over 3 years also received at 25 days of age 600-1200 ml of hyperimmune plasma from donors immunized with this vaccine, and as well at 4 days of age in foals with poor transfer of R. equi antibodies from their dams. The average foal mortality because of R. equi in the 380 foals annually to which hyperimmune plasma was administered dropped from 5.8% on these 3 farms to 0.2% (P < 0.05). Active vaccination of foals of unvaccinated mares on an enzootic farm at 20, 30, and 40 days of age did not protect them from mortality due to R. equi pneumonia. Serology was done by complement fixation and an agar gel immunodiffusion (AGID) tests using antigens prepared in the same manner as the vaccine antigens. The immune responses among hyperimmune plasma donors varied considerably as did the responses of vaccinated mares. Of 1117 serum samples with normal post suckling gammaglobulin levels (> 600 mg%) collected at 2 days of age from foals of vaccinated mares, 36% showed a negative or weak positive AGID reaction, while the remainder had positive to strongly positive reactions. PMID- 9226835 TI - Prevention of Rhodococcus equi pneumonia of foals using two different inactivated vaccines. AB - Two different, inactivated, aluminium salt adsorbed vaccines, one containing a R. equi strain (serotype 1, 10(9) CFU/ml and equine herpesvirus 2 (EHV-2) (1.5 x 10(7) PFU/ml) and another containing R. equi only were used on three studfarms to determine whether the disease can be prevented by vaccination of both pregnant mares and their foals. Pregnant mares received two 3 ml doses of vaccine intramuscularly 6 and 2 weeks before parturition and their foals were vaccinated on two or three occasions at 3, 5 or 7 weeks of age. The efficacy of the vaccines was evaluated on the basis of the clinical signs, serological response (indirect haemagglutination and virus neutralisation tests) and culture of R. equi from sick or dead foals. On studs A and B where the bivalent vaccine was used, 24 and 14 foals were born respectively to the vaccinated mares but no clinical case or death occurred due to R. equi pneumonia, while out of the 10 nonvaccinated control foals (stud B) two succumbed to R. equi pneumonia and 4 other foals had to be treated with antibiotics because of fever, coughing and dyspnea. In stud C, where the vaccine containing R. equi strain alone was used, all 15 vaccinated foals remained healthy but one of the 11 control foals died of suppurative R. equi pneumonia and one foal had to be treated due to R. equi pneumonia. R. equi strains (serotype 1) were isolated from the lungs of all dead foals. The serological response was very weak to both R. equi and the EHV-2 strain. Antibody titres in the colostrum of the vaccinated mares against R. equi (in studs A and B, geometric mean 3.79 +/- 1.63 and 4.14 +/- 1.46, respectively) were practically not higher than titres in the controls (in stud B geometric mean 2.12 +/- 1.96). More antibody was present in the colostrum samples against EHV-2 (geometric mean 6.1 + 1.4 compared to 2.5 +/- 1.2). In all foals antibody levels were hardly detectable against both R. equi and EHV-2 until five weeks of age. From the fifth week, antibody levels gradually increased and by the ninth week their reached a titre of 5.5 +/- 1.8 (2.7 +/- 1.2 in the control foals) against R. equi and 5.2 +/- 1.4 against EHV-2. The favorable clinical results and the low antibody titres in the sera of the vaccinated foals during the first week of life suggest that protection probably was due to repeated vaccination of young foals rather than to vaccination of mares. PMID- 9226836 TI - Assessment of the immunogenic potential of Rhodococcus equi virulence associated protein (VapA) in mice. AB - The development of immunity to Rhodococcus equi, particularly to a virulence associated protein (VapA) based antigen preparation, was examined in CD1 and BALB/c mice after intraperitoneal vaccination. Immunization with VapA based antigen without adjuvant markedly enhanced organ clearance in CD1 mice but not in BALB/c mice. Delayed type hypersensitivity response and antibody titres in VapA based antigen immunized BALB/c mice were less than in CD1 mice. By contrast also to CD1 mice, sera from immunized BALB/c mice did not react as strongly with VapA in western blots. Use of adjuvants (aluminium hydroxide, iscoms) interfered markedly with the immunogenic properties of the VapA based antigen, in the case of aluminium hydroxide by apparently driving a Th2 type of response. Unexpectedly, iscom adjuvants also impaired immunity and, despite the highest DTH response, produced a low IgG2a response, suggesting that iscomization of the antigen produced a low interferon gamma and high interleukin 2 response. Passive immunization of BALB/c mice with serum from mice immunized with live virulent strain 103+ resulted in only temporary and slight enhancement of organ clearance, supporting the central importance of cellular immunity to R. equi. Immunization with live virulence plasmid- and VapA-positive R. equi strain 103 resulted in marked liver clearance, in marked DTH response and high antibody titres. By contrast, immunization with live virulence plasmid- and VapA-negative strain 103 resulted in slight but variable enhancement of clearance, but insignificant DTH and antibody. The virulence plasmid, and by implication VapA, was thus shown to be critical in determining a highly effective protection to live organisms. PMID- 9226837 TI - Opsonic effect of equine plasma from different donors. AB - The ability of equine plasma from different donors to enhance phagocytic capacity was assessed in neutrophils obtained from seven foals, aged 7-8 days (Study A), and from seven adult horses (Study B). Neutrophils were allowed to phagocytize fluorescent yeast cells opsonized with plasma from one of three donors or with pooled serum, all previously frozen (-18 degrees C) and thawed. The results were analysed by flow cytometry. In study A, fresh autologous foal serum was also used for opsonization, and in study B, heat-inactivated plasma and pooled serum were used in addition to untreated samples. The plasma from donor GN induced a higher number of truly phagocytic neutrophils (mean 78%) than did plasma from donors GD (68%), OD (66%) and pooled serum (59%) when neutrophils from foals were used (p < 0.05). Similar results were obtained when adult neutrophils were used. Phagocytosis was markedly reduced with beat-inactivated plasma as a result of there being fewer phagocytic neutrophils and less phagocytized material per cell. The opsonic capacities of the autologous foal sera were lower than that of adult donor plasma in six out of seven foals. It is concluded that there is significant individual variation in the opsonic activity amongst plasma donors with similar serum IgG concentrations. The results were consistent irrespective of whether neutrophils from adults or foals were used. PMID- 9226838 TI - Immunophenotypic analysis of foal bronchoalveolar lavage lymphocytes. AB - The purpose of this study was to define the normal immunophenotype of equine lymphocytes present within the pulmonary air spaces, and to determine if this changes as foals age from one to ten weeks. Six pairs of mares and foals underwent sequential bronchoalveolar lavage (BAL) between 1 and 10 weeks of age. Data were grouped according to foal age (1, 1-3, 3-6, or 6-10 weeks of age) and were compared to adult control values obtained from the mares. BAL cells were harvested and stained with antibodies to the equine homologues of CD5, CD4, CD8, CD44, MHC I, MHC II and to equine IgG. Data, including percent positive staining and mean fluorescence intensity, were acquired on a flow cytometer gated for viable lymphocytes. All foals had significantly fewer CD5+ lymphocytes than mares, with the largest differences in the youngest animals. The percentage of CD4+ lymphocytes increased as the foals aged, approaching adult levels by 3 weeks of age, while the percentage of CD8+ lymphocytes increased more slowly and approached adult levels by 10 weeks of age. The CD4:CD8 ratio changed from 1.26 at one week of age to 0.78 by 10 weeks of age, compared to an adult value of 0.66. Lymphocytes from foals less than 6 weeks of age expressed MHC II and CD44 at lower levels than adults. The lymphocytic populations within the airways of foals are significantly different from adult animals. This may account for the susceptibility of foals to certain respiratory infections during the first few months of life. PMID- 9226839 TI - Antigenic analysis of Rhodococcus equi preparations using different horse sera. AB - An R. equi vaccine, prepared under conditions which induce the expression of many antigens, and which has given encouraging results in field trials, was analyzed by SDS-PAGE and immunoblots and compared with other R. equi preparations: a preparation made in with the same technique from a nonvirulent isolate (virulence associated protein negative, VapA-negative); a whole cell preparation of a VapA positive R. equi, prepared as a standard bacterin; and a semipurified VapA preparation (APTX). The antigens in these preparations were analyzed using hyperimmune sera (from adult horses vaccinated with the R. equi vaccine), passively and actively immunized foals' sera, asymptomatic but serologically positive foals' sera sera from R. equi pneumonic foals, an equine APTX antiserum, and a VapA monoclonal antibody (Mab). The vaccine under study had many proteins in high concentrations. Hyperimmune sera reacted strongly with vaccine antigens in the high molecular weight regions. In the low molecular weight range, it reacted in the 14 and less kDa zone. Sera from passively immunized foals reacted similarly but not so strongly. Actively immunized foals gave very weak reactions. With the APTX extract, the Mab reacted with bands at 15-17, 44 and 66 kDa; it reacted weakly with the whole cell and not with the VapA-negative preparations. The APTX antiserum and the Mab reacted strongly with the vaccine at the 14 and less kDa zone, and also with bands at 21, 44 and 66 kDa and very tenuously at 18 kDa, but not in the expected 15-17 kDa zone, suggesting that the native form of VapA is altered but without loss of antigenicity in the vaccine preparation. Our results suggest that other higher molecular weight antigens, in addition to VapA, may be important in inducing antibodies that protect young foals from R. equi pneumonia. These antigens are in high concentrations and in an immunogenic form in the vaccine. PMID- 9226840 TI - Pathogenesis and virulence of Rhodococcus equi. AB - Inhalation of the soil-borne organism, Rhodococcus equi, can lead to a chronic and severe pyogranulomatous pneumonia in young horses and immunocompromised people. In addition, ulcerative colitis is a common sequela to infection in foals, and dissemination from the lung to other body sites is not uncommon in either the horse or man. Although the facultative intracellular bacterium is susceptible to neutrophil-mediated killing, it is able to resist innate macrophage defenses and establish residence within the intracellular environment of that phagocyte. Definitive virulence factors of R. equi have not yet been determined, but potential candidates include capsular polysaccharide, the exoenzyme cholesterol oxidase, cell wall mycolic acids, and the products encoded by a virulence-associated plasmid. The ability to replicate within the macrophage is associated with virulence, and correlates in animals with the possession of a large plasmid and expression of the plasmid-encoded, surface-expressed lipoprotein, VapA. All strains of R. equi isolated from horses with clinical disease possess a large plasmid and express VapA antigens. In addition, bacterial clearance and granuloma development in mice is linked to plasmid possession and VapA expression. Plasmid containing strains replicate within the tissues of the mouse. whereas plasmid-cured strains are rapidly cleared. At present, the function of the VapA protein is unknown. In contrast to what is observed in the foal, only a small percentage of R. equi strains isolated from humans with rhodococcal disease express VapA antigens, although a high proportion of others express a related protein which is associated with reduced virulence and is also plasmid-encoded. In a limited number of plasmid-negative human isolates, virulence has been linked to beta-lactam resistance, and preliminary evidence suggests that the phenotype may be phage encoded. It is likely that the immune status of the patient can influence whether a particular strain of R. equi is able to produce clinical disease, and certainly experimental infection in mice has confirmed that an intact cellular immune response is necessary for clearance of the organism. PMID- 9226841 TI - Oxidation of macrophage membrane cholesterol by intracellular Rhodococcus equi. AB - Phagocytic uptake by cultured mouse macrophages (PD388D1) of a virulent strain (ATCC 33701) of Rhodococcus equi producing substantial cholesterol oxidase was accompanied by intracellular survival of the bacteria, and enzymatic oxidation of macrophage membrane cholesterol. A non-virulent strain (4219) lacking cholesterol oxidase was largely eliminated from the macrophages and did not bring about oxidation of membrane cholesterol. When R. equi 33701 was co-phagocytosed with Corynebacterium pseudotuberculosis there was a significant enhancement (10-fold) in the amount of oxidation product (4-cholesten-3-one) generated. R. equi and C. pseudotuberculosis are cooperative partners in the hemolysis of sheep erythrocytes, traceable to the cholesterol oxidase of the former, and phospholipase D of the latter. Results are discussed relative to the role of cooperative cytotoxins in damage to host tissue by bacterial pathogens. PMID- 9226842 TI - In vitro production of tumor necrosis factor-alpha, interleukin-6 and interleukin 8 from normal human peripheral blood mononuclear cells stimulated by Rhodococcus equi. AB - The capability of heat-killed Rhodococcus equi organisms to induce in vitro release of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6 and IL-8 from normal human mononuclear cells as well as the secretion kinetics of these inflammatory cytokines over a 48 h period were evaluated. Results show that normal human mononuclear cells are efficiently triggered to secrete TNF-alpha, IL 6 and IL-8 following R. equi stimulation according to a different kinetics. In particular, release of IL-B was already maximally expressed after 2 h of stimulation, while TNF-alpha amounts progressively increased in a time-dependent fashion. Finally, IL-6 secretion reached peak levels as soon as 18 h of incubation. Taken together, these data point out that monocyte-derived cytokines may play an important role in the immunological control of R. equi infection in immunocompetent people. PMID- 9226843 TI - Macroamphiphilic cell envelope components of Rhodococcus equi and closely related bacteria. AB - Recent progress towards an understanding of the architecture of the mycobacterial cell envelope (P.J. Brennan and H. Nikaido, Annual Review of Biochemistry 64 (1995) 29-63) provides a model with features more generally applicable to cell envelope organisation in other mycolic acid-containing bacteria. Using this archetype, a model for the organisation of the rhodococcal cell envelope is presented here, with particular reference to cell envelope composition in Rhodococcus equi. The likelihood that mycolic acids bound to the cell wall arabinogalactan contribute to the formation of a distinct outer lipid layer is emphasised. Furthermore, the model incorporates recent work which has characterised rhodococcal macroamphiphiles (lipoglycans and lipoproteins), including the VapA virulence-associated lipoproteins of R. equi. PMID- 9226844 TI - Pathogenicity and virulence of Rhodococcus equi in foals following intratracheal challenge. AB - Twelve foals, between 27 and 83 days old, were infected with 2 strains of Rhodococcus equi by intratracheal administration. Ten of the 12 foals were inoculated with 10(4)-10(10) colony forming units (cfu) of ATCC 33701 strain. The other 2 foals were inoculated with 10(9) cfu of a plasmid-cured derivative of the ATCC 33701 strain (ATCC 33701P-). All of the 10 foals challenged with the ATCC 33701 strain showed clinical signs of pulmonary disease within 5-13 days, such as gross lesions associated with acute bronchopneumonia and microscopic lesions associated with granulomatous pneumonia. The two foals challenged with the ATCC 33701P- strain showed neither clinical signs of disease nor gross lesions. Apparently, when lacking plasmid, the virulent Rhodococcus equi lost its pathogenicity. PMID- 9226845 TI - Clinical manifestations, diagnosis, treatment, and prevention of Rhodococcus equi infections in foals. AB - Since the 1986 Rhodococcus equi workshop, there have been major breakthroughs in understanding the epidemiology of, the virulence of, and the immune response to, this intriguing pathogen. However, with the exception of the use of hyperimmune plasma for the prevention of the disease (Martens et al., 1989; Madigan et al., 1991) the clinical aspects of R. equi infections have essentially remained unchanged. This article reviews the various clinical manifestations and summarizes recent advances in diagnosis, treatment and prevention of R. equi infections in foals. PMID- 9226846 TI - Comparison of tracheal aspiration with other tests for diagnosis of Rhodococcus equi pneumonia in foals. AB - The diagnostic value of tracheal aspiration was evaluated through comparison with other diagnostic methods using an experimental model of Rhodococcus equi (R. equi) pneumonia in foals. Pneumonia was induced by spraying of the virulent R. equi strain ATCC 33701 into the trachea of foals. All foals developed fever from 11 to 16 days after bacterial inoculation. One foal was euthanized on day 26 due to its poor prognosis, and other foals euthanized on day 43. During the experiment, some tests for diagnosis of Rhodococcus equi pneumonia such as tracheal aspiration, radiography, serodiagnosis and fecal culture were carried out. R. equi was continually isolated from tracheal aspirates collected via a silicone catheter inserted transnasally on day 8 to day 32 after bacterial inoculation. On the other hand, radiography, serodiagnosis and fecal culture were demonstrated to be valuable diagnostic methods, but to be limited compared with tracheal aspiration. Indirect fluorescent antibody technique (IFA) using a monoclonal antibody against the 15- to 17-kDa virulence-associated antigens (VapA) of R. equi and PCR targeting the structural gene of VapA detected bacteria in tracheal aspirates less sensitively than the isolation technique although they were more rapid. Therefore, we conclude that a combination of tracheal aspiration and bacterial isolation was the most valuable method for routine diagnosis of R. equi pneumonia in foals. PMID- 9226847 TI - Acute maduramicin toxicity in calves. AB - A herd of 277 beef-breed calves in three age groups was mistakenly given the poultry coccidiostat maduramicin in a total mixed ration. It caused an acute toxicosis in which sudden death was the sole clinical finding in most cases. One group of 212 calves aged five to eight months suffered a mortality of 51 per cent in eight days and a total mortality of 56 per cent during the 40 days in which mortality was recorded. Mortality of only 3 per cent was recorded in two other groups of calves aged nine to 16 months in eight days and a total mortality of 11 per cent over the 40-day period. PMID- 9226848 TI - Seroconversion in an industrial unit of rabbits infected with a non-pathogenic rabbit haemorrhagic disease-like virus. AB - A serological survey of 238 rabbits for antirabbit haemorrhagic disease virus (RHDV) antibodies was made in an industrial rabbitry where no signs of the disease had been reported for four years. Seroconversion was repeatedly detected and was due to a calicivirus antigenically related to RHDV but without its pathogenicity. There was a seroprevalence of 33.3 per cent among young animals at weaning at 31 days old, 27.6 per cent at five to seven days after weaning, 56.1 per cent at 13 to 14 days after weaning, 90.3 per cent at 19 to 20 days and 100 per cent at 32 to 33 days after weaning, and all the breeding rabbits were seropositive. In the last group and in the young at weaning, the anti-RHDV antibodies were mainly class IgG, but they were IgM and IgA at 13 to 14 days after weaning. In older fattening rabbits, there was a decrease of IgM and IgA and an increase of IgG confirmed seroconversion without any specific signs of rabbit haemorrhagic disease. On the basis of these results, the probable time of infection of the meat rabbits with this non-pathogenic virus was immediately after weaning. PMID- 9226849 TI - Cutaneous protothecosis in a dog. AB - A dog was infected systemically with Prototheca wickerhamii but showed only cutaneous protothecosis. The lesions appeared progressively and consisted of non pruritic scrotal swelling and ulceration, cutaneous nodules, crusty ulcerative lesions over the trunk and serous rhinitis. The diagnosis was based on skin biopsy findings and specific culture. Microscopic examination revealed a diffuse pyogranulomatous dermatitis and numerous protothecal organisms of different sizes within the cytoplasm of phagocytic cells. Treatment with oral ketoconazole for six months resolved all the clinical signs except the scrotal granuloma which, although it was significantly reduced, had to be removed surgically. However, after five months the condition returned. PMID- 9226850 TI - Pan-Asian spread of single fungal clone results in large scale fish kills. PMID- 9226851 TI - Postmortem comparison of ultrasonography, endocrine measurements and histology of large abnormal ovarian follicles in cows. PMID- 9226852 TI - An inherited neurological disorder of the St Bernard dog characterised by unusual cerebellar cortical dysplasia. PMID- 9226853 TI - Timing of cataract surgery in dogs. PMID- 9226854 TI - Arrangements for bovine abortion investigations. PMID- 9226855 TI - Gasterophilus pecorum larvae in an aged lion. PMID- 9226856 TI - Abdominal distension in a cat. PMID- 9226857 TI - Use of frontline spray in rabbits. PMID- 9226858 TI - Malposition of central venous catheters. Incidence, management and preventive practices. AB - INTRODUCTION: Proper placement is an essential prerequisite for the use of central venous catheters. Our study was undertaken to determine the incidence of aberrant locations dependent on different anatomic approaches for various types of central venous catheters and to elucidate failures and pitfalls of preventive practices. METHODS: 2580 percutaneously inserted lines (including 538 tunneled devices and 112 implantable Port-A-Caths) introduced by Seldinger's technique were reviewed for inadvertent malpositioning. RESULTS: Primary misplacement was evident on 47 occasions (1.82%), 38 times into large venous tributaries of the superior vena cava. 3 aberrant locations involved a persistent left superior vena cava, two catheters were placed into minor intrathoracic veins and in 3 patients inadvertent arterial cannulation occurred. The frequency of malpositioning was related to the anatomic approach and the catheter type used, but not to the physician's experience. Respective incidences were 4.12% for the left internal jugular access, but were lower for the right internal jugular (1.1%) and the right (1.01%) and left (0.89%) supraclavicular approach. Misplacement was more frequent with soft silicone catheters (2.53%) than with semi-rigid catheters (0.79%). All malpositions but one were detected on chest X-ray. DISCUSSION: Our data suggest that the incidence of catheter malposition depends on the site of insertion, the type of material used, but not on the experience of the physician who inserted the catheter. Scrupulous use of preventive practices reduces the frequency of malpositioned catheters, but physicians must keep in mind potential pitfalls. Injection of radioopaque contrast medium into the catheter during control chest X-ray should be done even with opaque catheters. PMID- 9226860 TI - Primary prevention mental health programs for children and adolescents: a meta analytic review. AB - Used meta-analysis to review 177 primary prevention programs designed to prevent behavioral and social problems in children and adolescents. Findings provide empirical support for further research and practice in primary prevention. Most categories of programs produced outcomes similar to or higher in magnitude than those obtained by many other established preventive and treatment interventions in the social sciences and medicine. Programs modifying the school environment, individually focused mental health promotion efforts, and attempts to help children negotiate stressful transitions yield significant mean effects ranging from 0.24 to 0.93. In practical terms, the average participant in a primary prevention program surpasses the performance of between 59% to 82% of those in a control group, and outcomes reflect an 8% to 46% difference in success rates favoring prevention groups. Most categories of programs had the dual benefit of significantly reducing problems and significantly increasing competencies. Priorities for future research include clearer specification of intervention procedures and program goals, assessment of program implementation, more follow up studies, and determining how characteristics of the intervention and participants relate to different outcomes. PMID- 9226861 TI - In praise of a cumulative prevention science. AB - Durlak and Wells (1997) provide a pivotal appraisal of prevention research on children and adolescents. Their meta-analytic approach has the advantages of reducing scientific misjudgments based on single studies, and providing a more balanced evaluation of impact of various interventions; it provides an opportunity for hypothesis finding helps set methodological standards, allows assessment of working classifications in the field, and an evaluation of the maturity of the prevention field itself. New developmental tasks for the field include incorporating and pursuing the leads produced by these findings, conducting similar research syntheses with other populations and outcomes, and using the results as an impetus to increased operational precision and parsimony. PMID- 9226862 TI - Life course development, community epidemiology, and preventive trials: a scientific structure for prevention research. PMID- 9226863 TI - An ounce of prevention research: what is it worth? PMID- 9226865 TI - One small step for science, one giant leap for prevention. PMID- 9226864 TI - A meta-analytic review of primary prevention programs for children and adolescents: contributions and caveats. PMID- 9226866 TI - A policy perspective on prevention. PMID- 9226867 TI - Meta-analysis of interventions to prevent mental health problems among youth: a public health commentary. PMID- 9226868 TI - Primary prevention mental health programs: the future is exciting. AB - Current outcome research on primary prevention mental health programs is encouraging and the future is exciting. Data continue to accumulate regarding the efficacy of preventive intervention. Exemplary programs can prevent multiple problems across different outcome domains suggesting the need for collaboration among preventionists across disciplines and research areas. The commentators on our review (Durlak and Wells, 1997) offered many useful suggestions to improve the next generation of research. Most recommendations fall broadly under the rubric of increasing the precision of theory, design, and program evaluation. If current recommendations for improving future research are followed, the next reviewers of primary prevention mental health programs for children and adolescents will have a more complete and useful database for analysis. PMID- 9226869 TI - Insertion sequence IST3091 of Thiobacillus ferrooxidans. AB - An insertion sequence, designated as IST3091, was located adjacent to the putative origin of replication region of plasmid pTFI91 of Thiobacillus ferrooxidans TFI-91. The DNA sequence of the transposase gene of IST3091 revealed similarity with that of IS30, IS1086, IS4351, and the integrase gene of SpV1-R8A2 B (a bacteriophage of Spiroplasma citri). The sequence of IST3091 is 1063 bp long with partially matched 30-bp terminal inverted repeats. Several restriction fragments of plasmid pTFI91 of T. ferrooxidans containing the IST3091 element were cloned into the vector pHSG398. The hybrid plasmids (pBTL) were transformed into Escherichia coli NK7379 containing a miniF plasmid, which was devoid of transposable elements. The transposition function of the IST3091 element was confirmed by mobilizing hybrid plasmids via conjugation from transformed E. coli NK7379 (donor) to E. coli M8820 (recipient). The presence of the transposed element in transconjugants was detected by polymerase chain reaction amplification. PMID- 9226870 TI - Soil isolates of Pseudomonas spp. that utilize inositol phosphates. AB - Soil bacteria that utilize inositol hexaphosphate (IHP) were isolated from a range of soils using defined selection media. An analysis of 200 randomly selected isolates indicated that less than 0.5% of the culturable population of soil bacteria were capable of using IHP as a sole source of C and P. From a further 238 isolates obtained from enrichment culture, four unique organisms (identified by randomly amplified polymorphic DNA-polymerase chain reaction) were selected and characterized for their ability to specifically utilize IHP. These four organisms were putatively identified as either fluorescent Pseudomonas spp. (P. putida CCAR53 and CCAR59) or nonfluorescent Pseudomonas spp. (P. mendocina CCAR31 and CCAR60) as determined by partial DNA sequence analysis of 16S rRNA genes. The fluorescent Pseudomonas strains exhibited marked phytase activity and liberated up to 81% of the phosphate from IHP either in the absence or presence of arabinose as an additional C source. The nonfluorescent strains also exhibited an ability to liberate Pi from IHP but were effective only in the presence of added arabinose. Strains CCAR59 and CCAR60 could effectively utilize either Na IHP or Ca-IHP at pH 7.0, whereas only strain CCAR59 could grow and utilize these substrates at pH 5.0. PMID- 9226871 TI - Use of Tn5-gusA5 to investigate environmental and nutritional effects on gene expression in the coronatine biosynthetic gene cluster of Pseudomonas syringae pv. glycinea. AB - Pseudomonas syringae pv. glycinea PG4180 produces coronatine (COR), a chlorosis inducing phytotoxin that consists of the polyketide coronafacic acid (CFA) coupled via an amide bond to the ethylcyclopropyl amino acid coronamic acid (CMA). Both CFA and CMA function as intermediates in the pathway to coronatine, and genes encoding their synthesis have been localized: however, the precise factors that regulate the production of COR and its precursors remain unclear. In the present study, a lambda delivery system for Tn5-gusA5 was developed and used to obtain transcriptional fusions in the COR gene cluster. Selected carbon (fructose and xylose) and amino acid (isoleucine and valine) sources significantly decreased COR biosynthesis at the transcriptional level. Transcriptional activity in the COR gene cluster was temperature dependent with maximal expression at 18-24 degrees C and significantly less expression at 14 and 30 degrees C. Interestingly, changes in osmolarity and the addition of complex carbon and nitrogen sources to the growth medium did not significantly affect COR gene expression, although both factors significantly impacted the quantity of COR produced. These results indicate that multiple factors impact COR production and only some of these directly affect transcription in the COR gene cluster. PMID- 9226872 TI - Comparison of partial 23S rDNA sequences from Rhizobium species. AB - A hypervariable region of Rhizobium 23S rDNA was amplified by polymerase chain reaction and phylogenetic relationships of several strains were determined by comparing nucleotide sequences of the amplified product. Variation in the 23S rDNA nucleotide sequences was consistent with phylogenetic relationships determined by host nodulation specificity and (or) 16S rDNA sequence analysis. Six strains representing three Rhizobium species (R. leguminosarum bv. trifolii, R. meliloti, and R. etli), and two strains each of Bradyrhizobium and Agrobacterium were clustered into five rDNA groups. Unique features identified by secondary structure analysis of the 23S rRNA sequenced region were consistent with the hypothesis that 23S rDNA could be used to design species- or strain specific Rhizobium probes. PMID- 9226873 TI - Recombinant plasmid mobilization between E. coli strains in seven sterile microcosms. AB - Transfer by mobilization of a pBR derivative recombinant plasmid lacking transfer functions (oriT+, tra-, mob-) from one E. coli K12 strain to another was investigated in seven sterile microcosms corresponding to different environments. These microcosms were chosen as representative of environments that genetically engineered microorganisms (GEMOs) encounter after accidental release, namely attached biomass in aquatic environments (biofilm), soil, seawater, freshwater, wastewater, mouse gut, and mussel gut, GEMOs survived in the same way as the host strains in all microcosms. Recombinant DNA mobilization occurred in the mouse gut, in sterile soil, and in biofilm. The plasmid transfer rates principally reflected the environmental conditions encountered in each microcosm. PMID- 9226874 TI - Correlation of Pseudomonas aeruginosa virulence factors from clinical and environmental isolates with pathogenicity in the neutropenic mouse. AB - The potential pathogenicity of a microorganism is a major concern for Health Canada evaluators, who will be processing new biotechnology products under the Canadian Environmental Protection Act. Potential pathogenicity is generally predicted by the results of animal pathogenicity studies. In an attempt to define surrogate data for an animal model, this study was initiated. Pseudomonas aeruginosa isolates from clinical and environmental sources were screened for their pilus type, serotype, lipopolysaccharide type, ability to evade host responses, and production of toxin A, exoenzyme S, elastase, phospholipase C, and total protease. The 50% lethal dose (LD50) of the same isolates was determined in the neutropenic mouse model of infection. An attempted correlation was drawn between each (or combinations) of the virulence determinants and the LD50. Stepwise linear regression showed that the presence of high levels of exoenzyme S in association with elastase or phospholipase C, or to a minor extent toxin A, was correlated with low numbers of bacteria required to elicit an LD50. No correlation between any of the other factors examined and virulence was detected. The data suggest that an in vitro high level of exoenzyme S production could be used as surrogate information for neutropenic mouse modelling; however, the levels of all of the extracellular enzymes should be considered when making such an assessment. PMID- 9226875 TI - Interaction between poly-3-hydroxybutyrate-co-3-hydroxyvalerate and a denitrifying Pseudomonas strain. AB - In a laboratory-scale system, dentrification activity of a heterotrophic microbial starter culture changed when different lots of poly-3-hydroxybutyrate co-3-hydroxyvalerate (P(HB-co-HV)) were used as the solid carbon source in the heterotrophic denitrification reactor. In this study, possible influences of physical and chemical properties of commercially produced P(HB-co-HV) (Biopol) on biofilm formation and metabolic activity of a denitrifying starter culture were investigated. These parameters indicate the polymers' suitability for the application as the matrix substance in the bioreactor. No differences in microstructure were detected between the different lots of polymers. Growth inhibitory effects by chemical additives were found in the case of triacetine, which was included as a plasticizer in seven of eight tested lots. The amount of hydroxyvaleric acid in the polymer was not assumed to affect denitrification activity. Relevant differences could be detected regarding primary adhesion of the starter culture Pseudomonas sp. strain 2nIII. It showed good adsorption properties to hydrophobic substances with a dependence on precultivation conditions. Pseudomonas sp. strain 2nIII degraded poly-3-hydroxybutyrate acid homopolymer and P(HB-co-HV) copolymers but was unable to break up poly-3 hydroxyvaleric acid. A possible reason for these findings is the substrate specifity of the polyhydroxyalkanoate depolymerase. PMID- 9226876 TI - Increased cellular fatty acid desaturation as a possible key factor in thermotolerance in Saccharomyces cerevisiae. AB - An increase of the unsaturation level of the cellular fatty acids was observed at sublethal or superoptimal temperatures in Saccharomyces cerevisiae. The hypothesis of this paper is that a high unsaturated fatty acids relative content "per se" is not a prerequisite for withstanding sublethal temperature stress in yeast but is the result of oxygen-consuming desaturase activation, with consequent reduction of oxygen and the oxygen free radicals as they form during thermal stress. In the thermotolerant strains, no increase of cellular thiobarbituric acid reactive substances (TBARSs) was observed when temperature approached the maximal growth temperature, suggesting prevention of oxidative damage. On the other hand, the values of TBARSs tripled at 42 degrees C in nonthermotolerant strains. When a sublethal hydrogen peroxide treatment preceded a rapid temperature rise, a selected thermotolerant strain responded with a relative increase of saturated fatty acids. This response, associated with an insignificant viability loss due to the double stress, suggests the induction an alternative oxygen consumption mechanism preventing excessive fatty acid unsaturation, which could be detrimental to the cells in the presence of hydrogen peroxide at sublethal temperatures. PMID- 9226877 TI - Purification, partial characterization and peptide sequences of vitellogenin from a reptile, the tuatara (Sphenodon punctatus). AB - Vitellogenin (Vg), a major precursor to egg yolk proteins, was purified from plasma of an estradiol-treated female tuatara (Sphenodon punctatus) by MgCl2-EDTA precipitation and DEAE-cellulose chromatography. The amino acid composition of tuatara Vg is similar to that of other vertebtate Vgs and contains a large proportion of serine (13.7 mol/100 mol of total amino acid). The amino acid sequences of the N-terminus of mature Vg (33 residues) and of several trypsin- and CNBr-generated peptides were determined. Six peptide sequences obtained from tuatara Vg could be aligned with Vg sequences from other vertebrates. Reduced and non-reduced forms of tuatara Vg have the same apparent molecular mass (approximately 218 kDa) when resolved by SDS-PAGE, indicating that inter-chain disulfide bonds are not a feature of the molecule in this species. Western blot analysis with anti-tuatara Vg antiserum indicated that at least some epitopes are shared among Vgs of turtle, alligator and tuatara. PMID- 9226878 TI - Stress response of lysosomal cysteine proteinases in rat C6 glioma cells. AB - Acid proteinases of C6 rat glioma cells were analyzed by means of gelatine polyacrylamide electrophoresis with respect to their responses to stress (heat shock and butanol). Proteinase activities on gelatine gels were characterized by their molecular masses. pH-optima, isoelectric points and reactions to inhibitors. Four bands of 25, 35 and 65/85 kDa most probably represent active and proforms as well as precursor complexes of lysosomal cysteine proteinases with pH optima between 4.0 and 5.0. The 25-kDa band seems to contain cathepsin L and B, the 35-kDa band proforms of cathepsin L and B and the 65/85-kDa bands possibly precursor complexes of cathepsin L and B. After 30-min heat shocks of different temperatures (40-50 degrees C), the 35-kDa activity increased, whereas the 65/85 kDa activity decreased after exposure to 42 and 44 degrees C, which also caused a strong increase in the level of the inducible heat shock protein of 68 kDa (HSP 68). The alterations of the proteinase activities and the increases of the HSP 68 levels occur at heat shock treatments that cause cell death in about 25-40% of the population as determined by Trypan blue staining. HSP 68 induction and proteinase activity changes were also observed 12 hr after a 1-hr treatment with different butanol concentrations (0.14-0.16 M). Kinetics of the response to a 30 min heat shock (44 degrees C) revealed a maximal decrease of the 35-kDa and a maximal increase of the 65/85-kDa activities after 12 hr recovery. When cells were exposed to repeated heat shocks (44 degrees C) at 12-hr intervals, the HSP 68 level further increased, whereas the 35-kDa and 65/85-kDa proteinase activities did not change. This result indicates a role of HSP 68 (or other HSPs) in the processing or stability of the putative cathepsin precursors (65/85-kDa complexes). PMID- 9226879 TI - Cloning and sequencing of a full-length sea bream (Sparus aurata) beta-actin cDNA. AB - A full-length cDNA clone encoding beta-actin (beta-actin) was isolated from a sea bream (Sparus aurata) liver cDNA library. Sequencing of this clone reveals an open reading frame encoding a 375 amino acid protein that shares a high degree of conservation to other known actins. The sea bream beta-actin sequence showed 98% identity to carp and human beta-actin and 95% and 94% identity to sea squirt and Dictyostelium cytoplasmic actins, respectively. PMID- 9226880 TI - Coordinate induction of tetrahydrobiopterin synthesis and nitric oxide synthase activity in chicken macrophages: upregulation of GTP-cyclohydrolase I activity. AB - Biosynthesis of nitric oxide (NO) and tetrahydrobiopterin (BH4) was investigated during cytokine-mediated activation of chicken macrophages. Monocyte derived macrophages and HD11 cells, a chicken macrophage cell line, constitutively synthesize BH4. Treatment of these cells with chicken macrophage activation factor (ChMAF) causes up to 10-fold increases of intracellular BH4 and of nitrite concentrations in the cell culture supernatant. Elevated BH4 levels correlate with an increase in GTP-cyclohydrolase I (GTP-CH) activity. Kinetic studies show a joint upregulation of GTP-CH activity and NO synthase activity first detectable 4 hr after stimulation. A corresponding increase in the mRNA for GTP-CH was detected by Northern blot analysis with a chicken GTP-CH specific cDNA probe. These results demonstrate that cytokine-induced BH4 synthesis by chicken macrophages is at least partially regulated through increased GTP-CH gene expression. The functional relevance of BH4 formation for NO production is shown by experiments using 2,4-diamino-6-hydroxypyrimidine (DAHP) as a specific inhibitor of GTP-CH. Monocyte derived macrophages stimulated in the presence of DAHP show a significant decrease in NO synthesis. The effect of DAHP was reversed by adding sepiapterin, which allows synthesis of BH4 through a salvage pathway. PMID- 9226881 TI - Effect of high ionic strength and inhibitors of H,K-ATPase on the ouabain sensitive K-p-nitrophenylphosphatase activity in the sea anemone Stichodactyla helianthus. AB - The ouabain-sensitive, K-stimulated p-nitrophenyl phosphatase (K-pNPPase) activity, an associated activity of the Na,K-ATPase, was assayed in tentacles of the sea anemone Stichodactyla helianthus to investigate the possibility that the sea anemone Na,K-ATPase activity is an associated activity of an H,K-ATPase. Activity was maximal at pH 6.5-7.0, decreasing only slightly in acidic medium but falling abruptly in alkaline medium to 60% of maximum at pH 7.4. The pH of maximum activity was not remarkably altered in high ionic strength medium (560 mM choline chloride), but ouabain-sensitive K-pNPPase activity of both rat and sea anemone was strongly inhibited. Inhibitors of the gastric H,K-ATPase, 100 microM omeprazole and 10 microM SCH 28080, did not inhibit the ouabain-sensitive K pNPPase activity. Activity of the sea anemone enzyme was inhibited by 10 microM ammonium vanadate, an inhibitor of P-type ATPase, and not by 2.5 mM sodium azide, an inhibitor of both F-type and V-type ATPase. Because the sea anemone K-pNPPase activity was previously found to be more sensitive to ouabain than the Na,K ATPase activity, K(+)-ouabain antagonism was investigated and found to be relatively muted, whereas K(+)-Na+ competition was stronger than in the rat kidney. PMID- 9226882 TI - Oxygen binding and aggregation of hemoglobin from the common European frog, Rana temporaria. AB - The hemoglobin from the European frog, Rana temporaria, consists of one major and three minor components. The tetramers aggregate upon deoxygenation notably at pH 7:3. Aggregation due to formation of disulphide bridges, as occurs in related species, was observed only in polyacrylamide gels. The hemolysate showed a pronounced Bohr effect. Oxygen affinity decreased with increasing hemoglobin concentration, indicating that aggregation affects the functional properties of the hemolysate. Oxygen binding equilibria of unfractionated hemolysate are insensitive to chloride and show low sensitivity to ATP. Analysis of oxygen equilibrium curves in terms of the two-state allosteric model (MWC) showed that pH change exerted a greater effect on the association constant of the oxygenated state (KR) than that of the deoxy state (KT). The number of interacting binding sites (q) increased with hemoglobin concentration. Cooperativity of oxygen binding, evaluated as Hill coefficient n, never exceeded the value of 3.0. Earlier studies on hemoglobin and blood from this and related species, report significantly higher n values at high O2 saturation. Molecular adaptive aspects are discussed. PMID- 9226883 TI - Baculovirus-mediated expression of chicken growth hormone. AB - A full-length chicken growth hormone (cGH) cDNA was placed downstream from the Autograph californica nuclear polyhedron virus, AcNPV, polyhedron gene promoter and expressed in Sf9 insect cells. Secreted recombinant cGH levels averaged 2-10 micrograms/ml from day 5-10 postinfection. The recombinant cGH analyzed by SDS PAGE gels and Western blotting consisted of a doublet with M(r) of 26.5 and 23.5 kDa. Analysis by 2-D electrophoresis of partially-purified recombinant cGH and purified native cGH revealed similar immunoreactive charge isoforms and M(r) variants. The recombinant hormone was biologically active in a homologous radioreceptor assay. The results show that cGH expressed in insect cells is biologically and immunologically active, and that a variety of isoforms are secreted which exhibit size and charge properties similar to those of pituitary derived cGH. PMID- 9226884 TI - Channelling of deoxyribose moiety of exogenous DNA into carbohydrate metabolism: role of deoxyriboaldolase. AB - In bacteria, the addition of (deoxy)nucleosides or (deoxy)ribose to the growth medium causes induction of enzymes involved in their catabolism, leading to the utilisation of the pentose moiety as carbon and energy source. In this respect, deoxyriboaldolase appears the key enzyme, allowing the utilisation of deoxyribose 5-P through glycolysis. We observed that not only deoxynucleosides, but also DNA added to the growth medium of Bacillus cereus induced deoxyriboaldolase; furthermore, the switch of the culture from aerobic to anaerobic conditions caused a further increase in enzyme activity, leading to a more efficient channelling of deoxyribose 5-P into glycolysis, probably as a response to the low energy yield of the sugar fermentation. In eukaryotes, the catabolism of (deoxy)nucleosides is well known. However, the research in this field has been mainly devoted to the salvage of the bases formed by the action of nucleoside phosphorylases, whereas the metabolic fate of the sugar moiety has been largely neglected. Our results indicate that the deoxyriboaldolase activity is present in the liver of several vertebrates and in a number of cell lines. We discuss our observations looking at the nucleic acids not only as informational molecules, but also as a not negligible source of readily usable phosphorylated sugar. PMID- 9226885 TI - Developmental changes in lipogenic enzymes in muscle compared to liver and extramuscular adipose tissues in the rabbit (Oryctolagus cuniculus). AB - The developmental changes in the activities of acetyl-CoA-carboxylase, malic enzyme and glucose-6-phosphate dehydrogenase were compared in longissimus muscle, liver and adipose tissues in growing rabbits. Activities of lipogenic enzymes were low in muscle, as compared to the other tissues studied. The lipogenic activities in longissimus muscle increased with age. This increase was well correlated with the age-related increase in intramuscular lipid content, suggesting that intramuscular adipose tissue results from in situ lipid synthesis. During growth, each tissue displayed a specific developmental pattern for lipogenic enzyme activities. In liver and adipose tissues, the three lipogenic enzyme activities first increased and subsequently decreased, during the postweaning period. In the muscle, no such decrease was observed, suggesting that intramuscular adipose tissue develops later than the other tissues tested. Throughout postnatal period, the ratio of malic enzyme to glucose-6-phosphate dehydrogenase was reversed in muscle compared to other fat sites (5 vs 0.04). Further studies are necessary to determine the role of malic enzyme in rabbit intramuscular lipid metabolism. PMID- 9226886 TI - A heterogeneous sialic acid-binding lectin with affinity for bacterial LPS from horse mussel (Modiolus modiolus) hemolymph. AB - A sialic acid-binding lectin that agglutinates a variety of erythrocytes and bacteria and react with sialoconjugates and purified lipopolysaccharides from marine vibrios has been affinity purified from hemolymph of the horse mussel Modiolus modiolus using Bovine submaxillary mucin conjugated to CNBr-activated Sepharose 4B. The lectin demonstrated heterogeneous activity, and at least two main entities were partially characterized, and are referred to as modiolin H and modiolin E activities for the agglutination of human and horse (equine) erythrocytes, respectively. Only modiolin E activity required calcium ions for hemagglutination. The M. modiolus lectin was mainly specific for NeuAc, although the lectin demonstrated a broader range of specificity, similarly to the Limulus polyphemus lectin. The purified lectin was a glycoprotein, and in the native state existed as aggregates with M(r) in the range of 100-1,300 kDa as observed by gradient-gel electrophoresis and gel filtration on Biogel and Superose. SDS PAGE under reducing conditions revealed three subunits of M(r) 14, 17.5 and 20 kDa. Various marine bacteria adsorbed the hemagglutinating activities of the M. modiolus lectin. Purified LPS preparations from various pathogenic marine vibrios were also effective inhibitors, in particular for modiolin E activity. These results indicate that the lectin play a role in recognition of bacteria. PMID- 9226887 TI - Microheterogeneity of odorant-binding proteins in the porcupine revealed by N terminal sequencing and mass spectrometry. AB - Several odorant-binding proteins (OBP) have been previously purified from the nasal mucosa of the old world porcupine Hystrix cristata. In this paper, we report their N-terminal amino-acid sequences and accurate molecular weights, as measured by electrospray mass spectrometry. The partial amino acid sequences reveal significant similarity with OBPs of other mammalian species and segregate the eight proteins purified into two subclasses. Mass spectrometry has revealed microheterogeneity among the proteins belonging to each of these two groups, suggesting a total number of OBPs of at least nine. The molecular weight differences between OBPs cannot be readily accounted for by common post translation modifications and indicate different gene products. Such a large number of different OBPs may represent further support to an odour discriminating role for these proteins. PMID- 9226888 TI - The 102 kDa chicken liver acid phosphatase: dimeric structure, glycoprotein nature and immunological properties. AB - In this study we isolated a highly purified, homogeneous high molecular weight (HMW) AcPase (Mr 102 kDa) from the chicken liver. This enzyme was shown to be a slightly acidic (pI 5.0-6.1), dimeric sialoglycoenzyme, composed of two equivalent subunits. Its sugar moiety, characterized by interactions with specific lectins, was shown to be composed of hybrid and complex type of carbohydrate chains. Heterogeneity of the high molecular weight AcPase arising from variations of the sugar components was demonstrated by isoelectric focusing, followed by reactions of the isoelectric components with specific lectins on NC membranes. Structural relationship based on immunological similarities was shown between the HMW AcPases from carp, frog, and chicken livers. PMID- 9226889 TI - Comparative biochemical analysis of sea urchin peristome and rat tail tendon collagen. AB - We report here a biochemical comparison between type 1 rat tail tendon collagen and collagen isolated from sea urchin peristome tissue. The sea urchin collagen consisted of two species of apparent mol masses, 140 and 116 kDa. Amino acid compositional analysis of the 140 and 116 kDa species revealed the presence of hydroxyproline and hydroxylysine as well as a glycine content of 28.1 mol.%. In solubility experiments the rat tail tendon collagen was found to precipitate at sodium chloride concentrations between 1 and 2 M while peristome collagen remained soluble at salt concentrations as high as 4 M. Incubation of the peristome and rat tail tendon collagen preparations with a sea urchin collagenase/gelatinase resulted in cleavage of the former but not the latter collagen. Upon heat denaturation at 60 degrees C, however, the rat tail tendon collagen served as a substrate for the gelatinase. Cyanogen bromide cleavage of rat tail and peristome collagens generated largely unique peptide maps. Collectively, these results suggest that structural differences exist between echinoderm and vertebrate type 1 collagens. PMID- 9226890 TI - Tsukamurella tyrosinosolvens sp. nov. AB - Chemotaxonomic and 16S ribosomal DNA sequence analyses of four bacterial isolates from blood cultures from patients with cardiac pacemaker implants and sputa of patients with chronic lung infections clearly demonstrated that these bacteria belong to the genus Tsukamurella. DNA-DNA hybridization data, as well as the physiological characteristics of the isolates, indicate that they are closely related and belong to a single species that differs from previously described members of the genus Tsukamurella. The name Tsukamurella tyrosinosolvens sp. nov. is proposed for these isolates, and the new species is represented by strain IMMIB D-1397T (= DSM 44234T). Strain IMMIB D-1397T exhibits 53.4, 53.5, and 54.7% DNA-DNA relatedness to Tsukamurella paurometabola DSM 20162T, Tsukamurella inchonensis DSM 44067T, and Tsukamurella pulmonis DSM 44142T, respectively. PMID- 9226891 TI - Thermococcus hydrothermalis sp. nov., a new hyperthermophilic archaeon isolated from a deep-sea hydrothermal vent. AB - An extremely thermophilic archaeon, strain AL662T, was isolated from a deep-sea hydrothermal vent located on the East Pacific Rise at a latitude of 21 degrees N. This strain is a strictly anaerobic coccus, and its cells range from 0.8 to 2 microns in diameter. The optimum temperature, pH, and Sea Salt concentration for growth are 85 degrees C, 6, and 20 to 40 g/liter, respectively. Strain AL662T grows preferentially on proteolysis products, on a mixture of 20 amino acids, and on maltose in the presence of elemental sulfur. The membrane lipids consist of di and tetraether glycerol lipids. The DNA G+C content is 58 mol%. Sequencing of the 16S rRNA gene showed that strain AL662T belongs to the genus Thermococcus. On the basis of hybridization results, we propose that this strain should be placed in a new species, Thermococcus hydrothermalis. PMID- 9226892 TI - Helicobacter rodentium sp. nov., a urease-negative Helicobacter species isolated from laboratory mice. AB - A spiral-shaped bacterium with bipolar, single, nonsheathed flagella was isolated from the intestines of laboratory mice. The organism grew at 37 and 42 degrees C under microaerobic and anaerobic conditions, did not hydrolyze urea, was weakly positive for catalase and oxidase, reduced nitrate to nitrite, did not hydrolyze indoxyl acetate or hippurate, and was resistant to cephalothin and nalidixic acid. This is the first urease-negative, murine Helicobacter spp. isolated from intestines. Also, Helicobacter pullorum and this bacterium are unique among the genus Helicobacter in having nonsheathed flagella. The new bacterium appears to be part of the normal intestinal flora; although its pathogenic potential is unknown, this organism was also isolated from scid mice with diarrhea that were co-infected with Helicobacter bilis. On the basis of 16S rRNA gene sequence analysis data and biochemical and phenotypic criteria, the new organism is classified as a novel helicobacter, for which we propose the name Helicobacter rodentium. The type strain is MIT 95-1707 (= ATCC 700285). PMID- 9226893 TI - Comparison of the 16S ribosomal DNA sequences from the intracellular agents of proliferative enteritis in a hamster, deer, and ostrich with the sequence of a porcine isolate of Lawsonia intracellularis. AB - Proliferative enteritis is an enteric disease that affects a variety of animals. The causative agent in swine has been determined to be an obligate intracellular bacterium, Lawsonia intracellularis, related to the sulfate-reducing bacterium Desulfovibrio desulfuricans. The intracellular agents found in the lesions of different animal species are antigenically similar. In addition, strains from the pig, ferret, and hamster have been shown to be genetically similar. In this study we performed a partial 16S ribosomal DNA sequence analysis on the intracellular agent of proliferative enteritis from a hamster, a deer, and an ostrich and compared these sequences to that of the porcine L. intracellularis isolate. Results of this study indicate that the intracellular agents from these species with proliferative enteritis have high sequence similarity, indicating that they are all in the genus Lawsonia and that they may also be the same species, L. intracellularis. PMID- 9226894 TI - Comparison of a new insertion element, IS1407, with established molecular markers for the characterization of Mycobacterium celatum. AB - Genomic analyses of 18 Mycobacterium celatum strains obtained from different patients in three countries (United States, United Kingdom, and France) were performed; the methods used in this study were restriction fragment length polymorphism (RFLP) analysis, pulsed-field gel electrophoresis (PFGE) analysis, and PCR restriction analysis (PRA) of the hsp-65 gene. A new insertion sequence, IS1407 (GenBank accession no. X97307), belonging to the IS256 family, was identified in M. celatum type 1 strains and was characterized by sequencing. When a probe for Mycobacterium xenopi IS1395-like sequences was used, the RFLP analysis of M. celatum type 1 strains revealed that they contained three or four copies of IS1407 in identical genomic positions, while this element was absent in all M. celatum type 2 strains. PFGE performed with three different endonucleases revealed a unique large restriction fragment (LRF) pattern for M. celatum type 1 strains, whereas the LRF patterns obtained for M. celatum type 2 strains were polymorphic. Moreover, PFGE of nondigested genomic DNA revealed extrachromosomal elements in M. celatum type 2. The type strain of M. celatum type 3 could not be differentiated from M. celatum type 1 strains on the basis of the results of the RFLP analysis, the PFGE analysis, and the PRA of IS1407. In this study we confirmed that M. celatum types 1 and 2 represent distinct genomic clusters and that the molecular markers in M. celatum type 2 exhibit greater polymorphism than the molecular markers in M. celatum type 1. PMID- 9226895 TI - Caloramator proteoclasticus sp. nov., a new moderately thermophilic anaerobic proteolytic bacterium. AB - A new moderately thermophilic proteolytic anaerobe, strain UT, was isolated from mesophilic granular methanogenic sludge. The cells were spore-forming, motile rods that were 0.4 micron wide and 2.4 to 4 microns long and stained gram negative. Electron micrographs of thin sections revealed the presence of an atypical gram-positive cell wall. Optimum growth occurred at 55 degrees C and at pH values between 7.0 and 7.5, with a doubling time of 30 min. The DNA base ratio of guanine plus cytosine was 31 mol%. The bacterium fermented proteins mainly to acetate, hydrogen, formate, and branched-chain fatty acids. Several amino acids, including glutamate, aspartate, arginine, histidine, threonine, methionine, and branched-chain amino acids, were also utilized. Glutamate was degraded to acetate, formate, hydrogen, and alanine. In addition, the strain degraded carbohydrates, including glucose, fructose, mannose, cellobiose, and starch, to acetate, ethanol, formate, lactate, and hydrogen. The results of a 16S rRNA sequence analysis phylogenetically placed strain UT in the low-guanine-plus cytosine-content subgroup of the gram-positive phylum. We propose to classify the described strain in the genus Caloramator as a new species, Caloramator proteoclasticus. The type strain of C. proteoclasticus, strain U, has been deposited in the Deutsche Sammlung von Mikroorganismen as strain DSM 10124. PMID- 9226896 TI - Reclassification of the crenarchael orders and families in accordance with 16S rRNA sequence data. AB - A phylogenetic analysis of all validly published members of the Crenarchaeota, including several new isolates from our laboratory, suggests three orders within this archaeal kingdom. The Thermoproteales consist of both the rod-shaped, hyperthermophilic, neutrophilic representatives of the Thermoproteaceae and the members of the new family Thermofilaceae. The Sulfolobales harbor all thermoacidophilic, coccoid organisms. The neutrophilic, hyperthermophilic cocci are members of a new order tentatively named "Igneococcales." This order comprises two families, the Desulfurococcaceae, characterized by maximal growth temperature of up to 100 degrees C, and the new family Pyrodictiaceae, for which optimal growth occurs at temperatures above 100 degrees C. PMID- 9226897 TI - Inter- and intraspecific genetic analysis of the genus Saccharomonospora with 16S to 23S ribosomal DNA (rDNA) and 23S to 5S rDNA internally transcribed spacer sequences. AB - In order to clarify interspecific relationships and to investigate the intraspecific phylogenetic structure of the genus Saccharomonospora, 16S to 23S ribosomal DNA (16S-23S) and 23S to 5S ribosomal DNA (23S-5S) internally transcribed spacers (ITSs) were used for sequence analyses. The 16S-23S and 23S 5S ITSs from 22 Saccharomonospora strains were amplified by PCR and directly sequenced. The average levels of nucleotide similarity of the 16S-23S and 23S-5S ITSs for the four valid species were 87.6% +/- 3.9% and 83% +/- 2.2%, respectively. For the most part, intraspecific sequence differences were not found in the two ITSs; the only exception was Saccharomonospora glauca K194, which differed from other S. glauca strains by 1 bp in the 23S-5S ITS. The Saccharomonospora viridis strains had a smaller 16S-23S ITS region than the other strains, which may be useful for differentiating these organisms from other Saccharomonospora species. The characteristics of the two ITS regions make them more useful than 16S rRNA sequences as a tool for defining and identifying Saccharomonospora strains. However, Saccharomonospora azurea K161T had two types of 23S-5S ITSs; rrnB, separated by XhoI digestion, had two additional nucleotides inserted between positions 52 and 55. Most of the 16S-23S and 23S-5S ITS sequences of S. azurea K161T and strains of "Saccharomonospora caesia" were identical; the only exception was rrnB in S. azurea K161T. The lengths and levels of sequence divergence of the two ITSs of Saccharomonospora sp. strain K180 were different from the lengths and levels of sequence divergence of the ITSs of other species. These findings suggest that a taxonomic revision of the genus Saccharomonospora is necessary. Two trees based on 16S-23S and 23S-5S ITS sequences revealed distinct interspecific relationships in the genus Saccharomonospora. PMID- 9226898 TI - Psychroserpens burtonensis gen. nov., sp. nov., and Gelidibacter algens gen. nov., sp. nov., psychrophilic bacteria isolated from antarctic lacustrine and sea ice habitats. AB - Psychrophilic, yellow-pigmented, seawater-requiring bacteria isolated from the pycnocline of meromictic Burton Lake and from sea ice cores obtained in the Vestfold Hills (68 degrees S, 78 degrees E) in eastern Antarctica were characterized. Phenotypic analysis showed that the strains isolated formed two distinct taxa. The first taxon included nonmotile, nutritionally fastidious strains that were isolated from the pycnocline of Burton Lake. The cells of these strains were morphologically variant, ranging from vibrioid to ring shaped to coiled and filamentous; in addition, the strains were unable to metabolise carbohydrates or polysaccharides and had DNA G + C contents of 27 to 29 mol%. The strains of the second taxon, which were isolated from sea ice cores and from ice aigal biomass, were saccharolytic, exhibited rapid gliding motility, were rodlike to filamentous, and had DNA G + C contents of 36 to 38 mol%. A 16S ribosomal DNA (rDNA) sequence analysis revealed that the two Antarctic taxa formed related but distinct lineages within the [Flexibacter] maritimus rRNA branch of the family Flavobacteriacrae. The levels of 16S rDNA sequence similarity between the taxa were 90.5 to 91.3%, while the levels of similarity to other members of the [F.] maritimus rRNA branch were 85 to 90%. The whole-cell lipid profiles of the Antarctic strains were mainly comprised of branched and unbranched monounsaturated C15 to C17 fatty acids. The presence of significant levels of the lipids a 15:1 omega 10c and a17:1 omega 7c appeared to be useful biomarkers for the new Antarctic taxa and for differentiating these organisms from other members of the family Flavobacteriaceae. On the basis of polyphasic taxonomic data we propose that the new taxa are novel bacterial species designated Psychroserpens burtonensis gen. nov., sp. nov. (type strain, ACAM 188) and Gelidibacter algens gen. nov., sp. nov. (type strain, ACAM 536). PMID- 9226899 TI - Use of ribotyping to distinguish Bordetella bronchiseptica isolates. AB - A total of 113 Bordetella bronchiseptica strains, isolated from 11 different host species worldwide, were characterized by ribotyping with restriction enzyme PvuII. Sixteen distinct ribotypes were identified, and each ribotype contained five to seven restriction fragments ranging in size from 1.8 to 5.6 kb. Approximately 88% of the swine isoaltes were identified as ribotype 3 strains. Isolates from dogs also displayed little variation; 74.1% were found to be ribotype 4 strains. Strains obtained from the remaining nine host species belonged to 15 different ribotypes. There was no association between geographic location and ribotype. The technique which we used may be useful for epidemiologic studies in which the transmission of B. bronchiseptica, both within and between species, is investigated. PMID- 9226900 TI - 16S rRNA gene sequence of Rubrobacter radiotolerans and its phylogenetic alignment with members of the genus Arthrobacter, gram-positive bacteria, and members of the family Deinococcaceae. AB - The nearly complete sequence of the 16S rRNA gene of an extremely highly radiotolerant bacterium, Rubrobacter radiotolerans (reclassified from Arthrobacter radiotolerans based on chemical characteristics), was determined by PCR amplification of the genomic DNA followed by cloning of the amplified gene and sequencing by the dideoxynucleotide method. The sequence was aligned with the sequences of members of the genus Arthrobacter and also with the sequences of representatives of the gram-positive bacteria having high G + C contents and the family Deinococcaceae (radioresistant micrococci and their relatives). The results of our phylogenetic analysis confirmed that R. radiotolerans is not a member of the Arthrobacter group and thus supported the previous reclassification. Moreover, although it is radioresistant and has a high G+C content, R. radiotolerans is more closely related to the gram-positive bacteria with high G+C contents than to the radioresistant members of the Deinococcaceae. PMID- 9226901 TI - Actinomyces europaeus sp. nov., isolated from human clinical specimens. AB - Ten strains of a hitherto undescribed catalase-negative, facultatively anaerobic, coryneform bacterium were isolated or collected by workers at three European clinical bacteriology laboratories or reference centers. These strains were isolated from humans, and most came from abscess material. Biochemical and chemotaxonomic characterization revealed that the strains belonged to the genus Actinomyces. The phenotypic features of the 10 strains were incompatible with the descriptions of the previously established Actinomyces species. A comparative 16S rRNA gene sequence analysis demonstrated that the previously undescribed strains constitute a new line in the genus Actinomyces. The name Actinomyces europaeus sp. nov. is proposed for these clinical isolates. The type strain is CCUG 32789A. PMID- 9226902 TI - rRNA sequences and evolutionary relationships among toxic and nontoxic cyanobacteria of the genus Microcystis. AB - A primary-structure analysis of the 16S rRNA gene was performed with 10 strains representing five described and one unidentified species of the genus Microcystis. The phylogenies determined illustrate the evolutionary affiliations among Microcystis strains, other cyanobacteria, and related plastids and bacteria. A cluster of 10 strains that included hepatotoxic isolates identified as Microcystis aeruginosa formed a monophyletic group. However, the genus Microcystis appeared to be polyphyletic and contained two strains that clustered with unicellular cyanobacteria belonging to the genus Synechococcus. The clustering of related Microcystis strains, including strains involved in the production of the cyclic peptide toxin microcystin, was consistent with cell morphology, gas vacuolation, and the low G + C contents of the genomes. The Microcystis lineage was also distinct from the lineage containing the unicellular genus Synechocystis and the filamentous, heterocyst-forming genus Nostoc. The secondary structure of a Microcystis 16S rRNA molecule was determined, and genus specific sequence signatures were used to design primers that permitted identification of the potentially toxic cyanobacteria belonging to the genus Microcystis via DNA amplification. PMID- 9226903 TI - Staphylococcus lutrae sp. nov., a new coagulase-positive species isolated from otters. AB - Phenotypic and phylogenetic studies were performed with three strains of a catalase-positive, gram-positive, coccus-shaped bacterium isolated from otters. The results of a 16S rRNA gene sequence analysis demonstrated that these strains represent a hitherto unknown subline within the genus Staphylococcus. Based on the results of the phylogenetic analysis and phenotypic criteria, we propose that these bacteria should be classified as members of a new species, Staphylococcus lutrae. The type strain of S. lutrae is DSM 10244. PMID- 9226904 TI - Amaricoccus gen. nov., a gram-negative coccus occurring in regular packages or tetrads, isolated from activated sludge biomass, and descriptions of Amaricoccus veronensis sp. nov., Amaricoccus tamworthensis sp. nov., Amaricoccus macauensis sp. nov., and Amaricoccus kaplicensis sp. nov. AB - Three isolates of gram-negative bacteria, strains Ben 102T, Ben 103T, and Ben 104T, were obtained in pure culture by micromanipulation from activated sludge biomass from wastewater treatment plants in Italy, Australia, and Macau, respectively. These isolates all had a distinctive morphology; the cells were cocci that usually were arranged in tetrads. Based on this criterion, they resembled other bacteria from activated sludge previously called "G" bacteria. On the basis of phenotypic characteristics and the results of 16S ribosomal DNA sequence analyses, the three isolates were very similar to each other, but were sufficiently different from their closest phylogenetic relatives (namely, the genera Rhodobacter, Rhodovulum, and Paracoccus in the alpha subdivision of the Proteobacteria) to be placed in a new genus, Amaricoccus gen. nov. Each of the three isolates represents a new species of the genus Amaricoccus; strains Ben 102T, Ben 103T, and Ben 104T are named Amaricoccus veronensis, Amaricoccus tamworthensis, and Amaricoccus macauensis, respectively. An isolate designated Ben 101T, which was isolated independently by Cech and Hartman in Kaplice, Czech Republic, was also characterized and belongs to the same genus. We propose that the isolate of Cech and Hartman should be placed in another new species, Amaricoccus kaplicensis. PMID- 9226905 TI - Bacillus salexigens sp. nov., a new moderately halophilic Bacillus species. AB - Bacillus salexigens sp. nov. is proposed based on the characteristics of six moderately halophilic, grampositive, rod-shaped strains isolated from salterns and hypersaline soils located in different geographical areas of Spain. These strains were motile, formed endospores, were strictly aerobic, were catalase and oxidase positive, and contained peptidoglycan of the meso-diamlnopimelic acid type in their vegetative cell walls. The DNA base compositions of these strains ranged from 36.3 to 39.5 mol%, and these organisms constitute a homology group with levels of DNA-DNA homology ranging from 73 to 100%. The 16S rRNA sequence of strain C-20MoT, which was used as the representative strain of these isolates, groups with the 16S rRNA sequences of members of the genus Bacillus, and the highest level of similarity is 95.4%. The type strain is strain C-20Mo (= ATCC 700290 = DSM 11483 = CCM 4646). PMID- 9226906 TI - Mycoplasma crocodyli sp. nov., a new species from crocodiles. AB - Organisms with the typical characteristics of mycoplasmas were isolated from joints and lungs of crocodiles. The results of growth inhibition tests and immunobinding assays showed that the 24 mycoplasma strains isolated were identical and distinct from previously described Mycoplasma, Entomoplasma, Mesoplasma, and Acholeplasma species. These organisms represent a new species, for which the name Mycoplasma crocodyli is proposed. M. crocodyli ferments glucose and maltose, does not produce films and spots, does not hydrolyze arginine, esculin, and urea, reduces tetrazolium chloride, and possesses phosphatase activity. It lyses and adsorbs bovine, ovine, and rabbit erythrocytes. Cholesterol or serum is required for growth. The optimum growth temperature is 37 degrees C. The G + C content of the DNA is 27.6 mol%. This organism causes exudative polyarthritis in crocodiles. The type strain of M. crocodyli is strain MP145 (= ATCC 51981). PMID- 9226907 TI - Streptomyces stramineus sp. nov., a new species of the verticillate streptomycetes. AB - Strain NRRL 12292T, which produces the bleomycin-like antibiotics LL-BO1208 alpha and LL-BO1208 beta, forms umbels consisting of chains of smooth-surface ovoid spores that are borne on verticils on the serial mycelia, indicating that it is a member of the verticillate group of the genus Streptomyces formerly classified in the genus Streptoverticillium. This strain was compared morphologically and physiologically to 54 other verticillate Streptomyces strains. The levels of DNA relatedness between strain NRRL 12292T and 34 other verticillate Streptomyces strains, including strains representing at least 19 genetic species clusters, were also determined. Strain NRRL 12292T is morphologically and physiologically distinct from the other verticillate strains studied, particularly because of the straw yellow color of its aerial mycelia and spore mass. DNA hybridization data support the uniqueness of this strain, since the levels of DNA relatedness between NRRL 12292T and the other verticillate strains used in this study were low. Our data support designation of a new species, Streptomyces stramineus, whose type strain is NRRL 12292. PMID- 9226908 TI - Desulfuromonas thiophila sp. nov., a new obligately sulfur-reducing bacterium from anoxic freshwater sediment. AB - A mesophilic, acetate-oxidizing, sulfur-reducing bacterium, strain NZ27T, was isolated from anoxic mud from a freshwater sulfur spring. The cells were ovoid, motile, and gram negative. In addition to acetate, the strain oxidized pyruvate, succinate, and fumarate. Sulfur flower could be replaced by polysulfide as an electron acceptor. Ferric nitrilotriacetic acid was reduced in the presence of pyruvate; however, this reduction did not sustain growth. These phenotypic characteristics suggested that strain NZ27T is affiliated with the genus Desulfuromonas. A phylogenetic analysis based on the results of comparative 16S ribosomal DNA sequencing confirmed that strain NZ27T belongs to the Desulfuromonas cluster in the recently proposed family "Geobacteracea" in the delta subgroup of the Proteobacteria. In addition, the results of DNA-DNA hybridization studies confirmed that strain NZ27T represents a novel species. Desulfuromonas thiophila, a name tentatively used in previous publication, is the name proposed for strain NZ27T in this paper. PMID- 9226909 TI - Spiroplasma platyhelix sp. nov., a new mollicute with unusual morphology and genome size from the dragonfly Pachydiplax longipennis. AB - Spiroplasma strain PALS-1T from the gut of the dragonfly Pachydiplax longipennis was shown to be distinct from other species, groups, and subgroups of the genus Spiroplasma as determined by reciprocal serological metabolism inhibition and deformation tests. However, this strain cross-reacted extensively with representatives of other groups when it was used as an antigen. Electron microscopy of cells of strain PALS-1T revealed cells surrounded by a single cytoplasmic membrane. Light microscopy revealed helical cells that exhibited twisting motility rather than rotatory or flexing motility. Variations in the tightness of coiling were transmitted from one end of the helix to the other. The strain was resistant to penicillin, which confirmed that no cell wall was present. The organism grew well in M1D and SP-4 liquid media under either aerobic or anaerobic conditions. Growth also occurred in 1% serum fraction medium and in conventional horse serum medium. The optimum temperature for growth was 30 degrees C, at which the doubling time was 6.4 h. Multiplication occurred at temperatures from 10 to 32 degrees C. Strain PALS-1T catabolized glucose and hydrolyzed arginine but not urea. The guanine-plus-cytosine content of the DNA was 29 +/- 1 mol%. The genome size was 780 kbp, the smallest genome size in the genus Spiroplasma. Strain PALS-1 (= ATCC 51748) is designated the type strain of a new species, Spiroplasma platyhelix. PMID- 9226910 TI - Sagittula stellata gen. nov., sp. nov., a lignin-transforming bacterium from a coastal environment. AB - A numerically important member of marine enrichment cultures prepared with lignin rich, pulp mill effluent was isolated. This bacterium was gram negative and rod shaped, did not form spores, and was strictly aerobic. The surfaces of its cells were covered by blebs or vesicles and polysaccharide fibrils. Each cell also had a holdfast structure at one pole. The cells formed rosettes and aggregates. During growth in the presence of lignocellulose or cellulose particles, cells attached to the surfaces of the particles. The bacterium utilized a variety of monosaccharides, disaccharides, amino acids, and volatile fatty acids for growth. It hydrolyzed cellulose, and synthetic lignin preparations were partially solubilized and mineralized. As determined by 16S rRNA analysis, the isolate was a member of the alpha subclass of the phylum Proteobacteria and was related to the genus Roseobacter. A signature secondary structure of the 16S rRNA is proposed. The guanine-plus-cytosine content of the genomic DNA was 65.0 mol%. On the basis of the results of 16S rRNA sequence and phenotypic characterizations, the isolate was sufficiently different to consider it a member of a new genus. Thus, a novel genus and species, Sagittula stellata, are proposed; the type strain is E-37 (= ATCC 700073). PMID- 9226911 TI - Bogoriella caseilytica gen. nov., sp. nov., a new alkaliphilic actinomycete from a soda lake in Africa. AB - A new gram-positive, alkaliphilic, nonsporulating, rod-shaped bacterium is described. The organism was isolated from soda soil (Lake Bogoria, Kenya) and has the following characteristics. It is nonmotile, not acid fast, halotolerant, and microaerophilic, and optimal growth occurs at pH values between 9 and 10. The peptidoglycan type is of type A4 alpha, with lysine as the characteristic diamino acid and glutamic acid as a component of the interpeptide bridge. The major menaquinone is MK-8(H4). The polar lipids are phosphatidylglycerol, diphosphatidylglycerol, phosphatidylinositol, and an unknown phospholipid. 12 Methyltetradecanoic acid is the predominant fatty acid. The G + C content of the DNA is 70 mol%. The results of 16S ribosomal DNA sequence comparisons revealed that strain HKI 0088T represents a new lineage in the order Actinomycetales. Therefore, we concluded that strain HKI 0088T should be assigned to a new genus and species, for which we propose the name Bogoriella caseilytica gen. nov., sp. nov. The type strain and only strain of this genus and species is HKI 0088 (= DSM 11294). PMID- 9226912 TI - Dethiosulfovibrio peptidovorans gen. nov., sp. nov., a new anaerobic, slightly halophilic, thiosulfate-reducing bacterium from corroding offshore oil wells. AB - A strictly anaerobic thiosulfate-reducing bacterium was isolated from a corroding offshore oil well in Congo and was designated strain SEBR 4207T. Pure culture of the strain induced a very active pitting corrosion of mild steel, with penetration rates of up to 4 mm per year. This constitutes the first experimental evidence of the involvement of thiosulfate reduction in microbial corrosion of steel. Strain SEBR 4207T cells were vibrios (3 to 5 by 1 microns), stained gram negative, and possessed lateral flagella. Spores were not detected. Optimum growth occurred in the presence of 3% NaCl at pH 7.0 and 42 degrees C. Strain SEBR 4207T utilized peptides and amino acids, but not sugars or fatty acids. It fermented serine, histidine, and Casamino Acids, whereas arginine, glutamate, leucine, isoleucine, alanine, valine, methionine, and asparagine were only used in the presence of thiosulfate. Peptides were fermented to acetate, isobutyrate, isovalerate, 2-methylbutyrate, H2, and CO2. The addition of either thiosulfate or sulfur but not sulfate increased peptide utilization, growth rate, and biomass; during growth, H2S was produced and a concomitant decrease in H2 was observed. The addition of either thiosulfate or sulfur also reversed H2 inhibition. 16S rRNA sequence analysis indicates that strain SEBR 4207T is distantly related to members of the genus Thermoanaerobacter (83% similarity). Because the phenotypic and phylogenetic characteristics cannot be assigned to any described genus, strain SEBR 4207T is designated as a new species of a new genus, Dethiosulfovibrio peptidovorans gen. nov., sp. nov. Strain SEBR 4207T has been deposited in the Deutsche Sammlung von Mikroorganismen und zellkulturen GmbH (= DSM 11002). PMID- 9226913 TI - Propionibacterium cyclohexanicum sp. nov., a new acid-tolerant omega-cyclohexyl fatty acid-containing propionibacterium isolated from spoiled orange juice. AB - A non-spore-forming, coryneform bacterium, strain TA-12T, was isolated from spoiled off-flavor orange juice. Growth of this organism occurs at pH 3.2 to 7.5, and optimum growth occurs at pH values between 5.5 and 6.5. This organism produces lactic acid, propionic acid, and acetic acid from glucose. It is catalase negative. The cells are heat resistant and can withstand a temperature of 90 degrees C for 10 min. The DNA G + C content is 66.8 mol%. This strain has as MK-9(H4) respiratory quinone system and contains meso-diaminopimelic acid in its cell wall, and omega-cyclohexyl undecanoic acid is the major cellular fatty acid. The results of a phylogenetic analysis of the 168 rRNA gene of this organism indicated that its highest level of homology is its level of homology with the representative of the classical propionibacteria, Propionibacterium freudenreichii (97.1%). Strain TA-12T is phenotypically similar to P. freudenreichii, but it produces a large amount of lactic acid and has a distinct fatty acid composition, acid tolerance, and heat resistance, which differentiate it from P. freudenreichii and other propionic acid-producing bacteria. On the basis of these findings we propose the name Propionibacterium cyclohexanicum sp. nov. for this organism. The type strain is TA-12 (= IAM 14535 = NRIC 0247). PMID- 9226914 TI - Tetragenococcus muriaticus sp. nov., a new moderately halophilic lactic acid bacterium isolated from fermented squid liver sauce. AB - A total of 11 strains of moderately halophilic histamine-producing bacteria isolated from fermented squid liver sauce were studied phenotypically, genotypically, and phylogenetically. These strains are considered members of the genus Tetragenococcus based on their physiological, morphological, and chemotaxonomic characteristics. A 16S rRNA gene sequence analysis showed that these strains clustered with, but were separate from, Tetragenococcus halophilus. The results of DNA-DNA hybridization experiments indicated that the new isolates represent a new Tetragenococcus species, for which we propose the name Tetragenococcus muriaticus; strain X-1 (= JCM 10006) is the type strain of this species. PMID- 9226915 TI - Phylogenetic relationship of the twenty-one DNA groups of the genus Acinetobacter as revealed by 16S ribosomal DNA sequence analysis. AB - The inter- and intrageneric relationships of members of the genus Acinetobacter were investigated by performing a comparative sequence analysis of PCR-amplified 16S ribosomal DNAs (rDNAs) from 21 strains representing all of the DNA groups that have been described. Phylogenetic treeing confirmed that Acinetobacter spp. form a coherent cluster within the gamma subdivision of the class Proteobacteria that includes strains with overall levels of 16S rDNA sequence similarity of more than 94%. The analysis of intrageneric relationships suggested that the majority of the strains cluster in five clearly distinguishable clusters, and this conclusion was supported by the results obtained with the different methods used for phylogenetic analysis (i.e., the maximum-likelihood, parsimony, and distance matrix methods). The first cluster contains the representatives of DNA groups 2 (Acinetobacter baumannii) and TU13, whereas the second cluster comprises representatives of DNA groups 3, "Close To TU13," and "between 1 and 3." The representatives of closely related Acinetobacter DNA groups 8 (Acinetobacter twoffii) and 9 belong to the third cluster, which includes the representative of DNA group 6 as well. The fourth cluster is formed by DNA groups BJ15, BJ16, and BJ17, and the fifth cluster comprises DNA groups 1 (Acinetobacter calcoaceticus), BJ14, 10, and 11. Within the fifth cluster the 16S rDNA sequences of DNA group 10 and 11 strains are nearly identical. The representatives of DNA groups 4 (Acinetobacter haemolyticus), 5 (Acinetobacter junii), 7 (Acinetobacter johnsonii), 12 (Acinetobacter radioresistens), TU14, and TU15 form individual branches that are not significantly affiliated with any of the five clusters identified. Apart from the clustering of the most closely related DNA groups, the general topology of the distance dendrogram revealed some discrepancy with previous DNA-DNA hybridization data, which may point to the inadequacy of comparative 16S rDNA sequence analysis for reflecting true evolutionary relationships of closely related bacterial taxa. Important, however, was the presence of unique sequence motifs in each of the 21 different DNA groups studied, which may be useful for rapid differentiation of DNA groups of the genus Acinetobacter. PMID- 9226916 TI - Treponema amylovorum sp. nov., a saccharolytic spirochete of medium size isolated from an advanced human periodontal lesion. AB - A highly motile, medium-size, saccharolytic spirochete was isolated from an advanced human periodontal lesion in medium OMIZ-Pat supplemented with 1% human serum. The growth of this organism is dependent on either glucose, maltose, starch, or glycogen. The cells contain six endoflagella, three per pole, which overlap in the central region of the cell body. On the basis of its cell morphology and enzyme activities, as well as its sodium dodecyl sulfate polyacrylamide gel electrophoresis protein and antigen profiles, this organism is clearly distinct from all previously cultured spirochetes. The presence of a novel species is supported by the 16S rRNA sequence of this organism, which places it in phylotype 19 of Choi et al. (B. K. Choi, B. J. Paster, F. E. Dewhirst, and U. B. Gobel, Infect. Immun. 62:1889-1895, 1994). The only isolate, strain HA2P, is designated the type strain of a novel species, for which we propose the name Treponema amylovorum. PMID- 9226917 TI - Pseudomonas monteilii sp. nov., isolated from clinical specimens. AB - We propose the name Pseudomonas monteilii for a new species of gram-negative, rod shaped, motile bacteria that were nonhemolytic on blood agar and were isolated from clinical sources. The 10 strains of P. monteilii were incapable of liquefing gelatin. They grew at 10 degrees C but not at 41 degrees C, produced fluorescent pigments, catalase, and cytochrome oxidase, and possessed the arginine dihydrolase system. They were capable of respiratory but not fermentative metabolism. They did not hydrolyze esculin or starch and were able to use benzylamine, alpha-aminobutyrate, D-ribose, L-arabinose, butyrate, valerate, isovalerate, isobutyrate, inositol, phenylacetate, D-alanine, and amylamine. They possessed L-phenylalanine arylamidase, L-lysine arylamidase, L-alanine arylamidase, gamma-glutamyl-transferase, glycyl-phenylalanine arylamidase, L tryptophan arylamidase, glycyl-L-alanine arylamidase, esterase C4, esterase C6, esterase C8, esterase C9, esterase C10, and esterase C18. DNA relatedness studies revealed that P. monteilii strains formed a homogeneous DNA hybridization group. A total of 57 strains representing previously described or partially characterized taxa belonging to the genus Pseudomonas were 6 to 54% related to P. monteilii. The highest hybridization values were obtained with strains belonging to or related to Pseudomonas putida biovar A. The average G+C content of the DNA was 60.5 +/- 0.5 mol% for four of the P. monteilii strains studied. The type strain of P. monteilii is CFML 90-60 (= CIP 104883); it was isolated from bronchial aspirate and has a G+C content of 60 mol%. The clinical significance of these organisms is not known. PMID- 9226918 TI - Diversity of alkaliphilic halobacteria: proposals for transfer of Natronobacterium vacuolatum, Natronobacterium magadii, and Natronobacterium pharaonis to Halorubrum, Natrialba, and Natronomonas gen. nov., respectively, as Halorubrum vacuolatum comb. nov., Natrialba magadii comb. nov., and Natronomonas pharaonis comb. nov., respectively. AB - The 16S rRNA genes of three species of the genus Natronobacterium (Natronobacterium gregoryi, Natronobacterium pharaonis, and Natronobacterium vacuolatum) were sequenced and compared to that of the previously sequenced species Natronobacterium magadii. The sequences revealed that Natronobacterium pharaonis was phylogenetically distinct from the other members of the genus and also from other recognized genera of the family Halobacteriaceae. However, Natronobacterium vacuolatum and Natronobacterium magadii were found to be most closely related to the genera Halorubrum and Natrialba, respectively. An unidentified haloalkaliphile, strain SSL1, was also closely related to Natronobacterium magadii and Natrialba asiatica. On the basis of phylogenetic tree reconstructions, signature bases specific for individual genera, and sequences of spacer regions between 16 and 23S rRNA genes, we propose the following changes: Natronobacterium pharaonis to be transferred to Natronomonas gen. nov. as Natronomonas pharaonis gen. nov., comb. nov.; Natronobacterium vacuolatum to be transferred to the genus Halorubrum as Halorubrum vacuolatum comb. nov.; and Natronobacterium magadii to be transferred to the genus Natrialba as Natrialba magadii. PMID- 9226919 TI - Inter- and intraspecies comparison of the 16S-23S rRNA operon intergenic spacer regions of six Listeria spp. AB - The 16S-23S rRNA intergenic spacer (IGS) regions found in six Listeria species were characterized. PCR amplification of the 16S-23S IGS with a "generic primer" set generated products of about 340 bp (small) and 550 to 590 bp (large) with DNA from all Listeria strains tested. Seven Listeria monocytogenes serotype 4b strains and one L. monocytogenes serotype 4d strain also had an additional PCR product of ca. 360 bp. The 360-bp PCR product from one of these L. monocytogenes serotype 4b strains was identical in nucleotide sequence to the small 340-bp IGS, except that it contained an 18-bp tandem repeat. The small rRNA IGSs of L. innocua, L. ivanovii, L. seeligeri, L. welshimeri, and L. grayi were 83 to 99% homologous to that of L. monocytogenes. The large rRNA IGS of L. monocytogenes was 81 to 96% homologous to those of the other Listeria species and agreed with current taxonomic division among these species. The nucleotide sequences of the central 274 bp of the large rRNA IGS of strains from seven different L. monocytogenes serotypes were highly homologous; however, serotype-specific differences were noted, and four groups were identified within L. monocytogenes based on this analysis. PMID- 9226920 TI - Rhizobium hainanense sp. nov., isolated from tropical legumes. AB - A fast-growing rhizobial group isolated from leguminous plants in Hainan Province, a tropical region of China, is proposed as a new Rhizobium species on the basis of 16S rRNA gene sequencing. DNA-DNA hybridization, and phenotypic characterization. This new species belongs to the phylogenetic branch which includes Rhizobium leguminosarum. We propose the name Rhizobium hainanense sp. nov. for this species. The strain CCBAU 57015 (166) is the type strain; it has been deposited in the culture collection of Beijing Agricultural University, People's Republic of China. PMID- 9226921 TI - Phylogenetic and genetic relationships of Mesorhizobium tianshanense and related rhizobia. AB - The genetic and phylogenetic relationships for strains of Mesorhizobium tianshanense and its relatives were compared by an analysis of the results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of whole cell proteins, DNA-DNA hybridization, and full 16S rRNA gene sequencing. The strains of M. tianshanense formed a cluster which was distinct from those of other rhizobium species in the clustering analysis of SDS-PAGE. DNA-DNA relatedness between A-1BS (type strain of M. tianshanense) and the type or reference strains for Mesorhizobium loti, M. huakuii, M. ciceri, M. mediterraneum, and cluster U, an unnamed rhizobial group, ranged from 4.4 to 43.8%. The phylogenetic analysis based on the 16S rRNA gene sequences showed that M. tianshanense was closely related to the Mesorhizobium phylogenetic branch and could be distinguished from the other four species in this branch. These results further confirmed that these bacteria constitute a distinct rhizobial species. PMID- 9226922 TI - Phenotypic and phylogenetic characterization of some Globicatella-like organisms from human sources: description of Facklamia hominis gen. nov., sp. nov. AB - Six strains of a hitherto undescribed gram-positive, catalase-negative, facultatively anaerobic coccus from human sources were characterized by phenotypic and molecular taxonomic methods. Comparative 16S rRNA gene sequencing studies demonstrated that the unknown strains were genealogically homogeneous and constitute a new line closely related to, but distinct from, the genus Globicatella. The unknown bacterium was readily distinguished from Globicatella sanguis, the type species of the genus Globicatella, by the results of biochemical tests and an electrophoretic analysis of whole-cell proteins. Based on phylogenetic and phenotypic evidence, we propose that the unknown bacterium be classified as Facklamia hominis gen. nov., sp. nov. The type strain of Facklamia hominis is CCUG 36813. PMID- 9226923 TI - Identification of rickettsiae isolated in Japan as Coxiella burnetii by 16S rRNA sequencing. AB - The 16S rRNA genes of Japanese Coxiella isolates obtained from various sources and geographical areas were directly sequenced by dideoxynucleotide chain termination methods in which Taq DNA polymerase was used. The levels of sequence similarity among Japanese, European, and American isolates were more than 99%, and the Japanese isolates were identified as Coxiella burnetii, C. burnetii strains isolated worldwide, including Japan, were found to be very similar. PMID- 9226924 TI - Actinomyces graevenitzii sp. nov., isolated from human clinical specimens. AB - Four strains of a previously unknown, catalase-negative, facultatively anaerobic, gram-positive, rod-shaped organism originating from humans were characterized by biochemical, chemical, and molecular taxonomic methods. The four strains phenotypically closely resembled one another, and although they possessed characteristics consistent with membership in the genus Actinomyces, they differed from all previously recognized species of this genus. The results of comparative 16S rRNA gene sequencing studies demonstrated that the unknown human bacterium was phylogenetically a member of the genus Actinomyces. Within the genus Actinomyces, the unidentified bacterium formed a loose, but statistically significant, association with a subgroup which included Actinomyces bovis, the type species of the genus. 16S rRNA sequence divergence values of > 6%, however, unequivocally demonstrated that the unidentified bacterium represents a new subline of the genus Actinomyces. A new species, Actinomyces graevenitzii, is proposed for the four new isolates. The type strain of A. graevenitzii is CCUG 27294. PMID- 9226925 TI - The tannin-degrading species Streptococcus gallolyticus and Streptococcus caprinus are subjective synonyms. AB - The tannin-degrading species Streptococcus gallolyticus and Streptococcus caprinus have been shown to be subjective synonyms on the basis of their levels of 16S rRNA sequence similarity (98.3%) and DNA-DNA homology (> 70%) and the phenotypes of their type strains. S. gallolyticus has nomenclatural priority according to Rule 24b(2) of the International Code of Nomenclature of Bacteria. PMID- 9226926 TI - Characterization of some Actinomyces-like isolates from human clinical specimens: reclassification of Actinomyces suis (Soltys and Spratling) as Actinobaculum suis comb. nov. and description of Actinobaculum schaalii sp. nov. AB - Five strains of a hitherto unknown Actinomyces-like bacterium were isolated from human clinical sources, including blood cultures. Biochemical and chemotaxonomic characterization indicated that the strains were distinct from previously described Actinomyces and Arcanobacterium species. A comparative 16S rRNA gene sequence analysis demonstrated that the undescribed strains constitute a new subline within the Actinomyces-Arcanobacterium species complex. The closest known relative of the isolates was found to be Actinomyces suis, although a 16S rRNA sequence divergence value of approximately 6% clearly demonstrated that the unknown bacterium represents a distinct species. Based on the results of the present and earlier phylogenetic investigations, it is proposed that Actinomyces suis should be reclassified in a new genus, the genus Actinobaculum, as Actinobaculum suis comb. nov. In addition, a new species, Actinobaculum schaalii, is proposed for the Actinomyces-like bacterium from human sources. The type strain of Actinobaculum schaalii is CCUG 27420. PMID- 9226927 TI - Reclassification of Nocardioides simplex ATCC 13260, ATCC 19565, and ATCC 19566 as Rhodococcus erythropolis. AB - Our phylogenetic analysis based on 16S ribosomal DNA (rDNA) sequences and chemotaxonomic analyses showed that Nocardioides simplex ATCC 13260, ATCC 19565, and ATCC 19566 are more closely related to the genus Rhodococcus, especially Rhodococcus erythropolis, than to the genus Nocardioides. N. simplex ATCC 13260 and N. simplex ATCC 19565 and ATCC 19566 exhibited levels of 16S rDNA similarity of 99.4 and 100%, respectively, to R. erythropolis DSM 43066T. Strains ATCC 13260, ATCC 19565, and ATCC 19566 had mesodiaminopimelic acid in their peptidoglycan and MK-8(H2) as their predominant menaquinone. These three strains produced cellular fatty acid patterns similar to those of R. erythropolis strains rather than those of Nocardioides species. Therefore, N. simplex ATCC 13260, ATCC 19565, and ATCC 19566 should be reclassified as strains of R. erythropolis Gray and Thornton 1928. PMID- 9226928 TI - Characterisation of human serum albumin heterogeneity by capillary zone electrophoresis and electrospray ionization mass spectrometry. AB - Human serum albumin (HSA) preparations and HSA-containing recombinant human erythropoietin (rhEPO) formulations were analyzed by capillary zone electrophoresis. HSA was separated into several components by the addition of 1,4 diaminobutane to 20 mM sodium phosphate, pH 6.0 as electrophoretic buffer. Resolution was improved by increasing the buffer pH to 8.5. A comparative analysis of HSA preparations from three manufacturers provided evidence that this method can be used to differentiate individual preparations. The analysis of rhEPO formulations, composed largely of HSA, showed levels of heterogeneity comparable to that of HSA preparations. Electrospray ionisation mass spectrometry was used as an independent method to confirm the heterogeneous nature of HSA. PMID- 9226929 TI - Direct determination of procainamide and N-acetylprocainamide by capillary zone electrophoresis in pharmaceutical formulations and urine. AB - In this work a new sensitive capillary zone electrophoresis method for the direct determination of procainamide (PA) and N-acetylprocainamide (NAPA) in pharmaceutical formulations and urine samples without any extraction and/or preconcentration steps has been developed. The determination was carried out in a fused-silica capillary of 43.5 cm (35.9 cm length to the detector) x 0.75 micron J.D. Phosphate 0.05 M buffer was used as the background electrolyte and 10 kV separation voltage was applied. The separation of PA and NAPA is possible in a wide range of pH from 1.7 to 9.7. However, in order to avoid the effect of the urine matrix, it is optimal to work at pH 7.7. The determination of PA and NAPA takes less than 5 min while high resolution is achieved. The detection limits obtained, 1.235 micrograms/ml and 0.359 microgram/ml for PA and NAPA respectively, are lower than those for GC method normally reported. PMID- 9226930 TI - Identification of alpha 1-acid glycoprotein (orosomucoid) in human synovial fluid by capillary electrophoresis. AB - Capillary electrophoresis of human synovial fluid in a phosphate borate run buffer containing sodium dodecyl sulphate separates a hydrophilic glycoprotein, hyaluronan and a number of low-molecular-mass components. The hydrophilic glycoprotein is identified as alpha 1-acid glycoprotein (AGP), orosomucoid, by co injection methods with human AGP and by reaction with neuraminidase which released N-acetylneuraminic acid. Finally, a sample of the glycoprotein was isolated by micropreparative capillary electrophoresis, examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis methods and shown to give a positive reaction with AGP antibodies. The peak due to AGP in the capillary electrophoresis is broad and gives evidence for the presence of glycoforms. PMID- 9226931 TI - Functional equivalence in children: derived stimulus-response and stimulus stimulus relations. AB - The present study investigated the simultaneous occurrence of emergent stimulus response relations (functional equivalence) and stimulus-stimulus relations (stimulus equivalence). After being pretrained and tested on two symbolic match to-sample tasks (X1-Y1, X2-Y2), 20 4- and 5-year-old children were trained to emit specified responses to pairs of stimuli (A1-R1, B1-R1, A2-R2, B2-R2) in one setting (original training) and to emit other responses to one member of each pair (A1-R3, A2-R4) in another setting (reassignment training). Of the 18 children who responded correctly on all trained tasks, 15 emitted the novel responses also in the presence of the nonreassigned stimuli (B1-R3, B2-R4). Eleven of these children also matched same-class stimuli with one another (A1-B1, A2-B2, and vice versa). Additional tests with four of these children documented the formation of conditional response-stimulus relations (R3-B1, R4-B2) in all four children, and of conditional response-response relations (R1-R3, R2-R4, and vice versa) in two of them. Children who did not show stimulus control transfer also failed to match same-class stimuli with one another. Present findings, together with those obtained in animal research, suggest that functional equivalence can imply but does not require stimulus equivalence. PMID- 9226932 TI - Phonological awareness deficits in developmental dyslexia and the phonological representations hypothesis. AB - The claim that the well-documented difficulties shown by dyslexic children in phonological awareness tasks may arise from deficits in the accuracy and the segmental organization of the phonological representations of words in their mental lexicons is receiving increasing interest from researchers. In this experiment, two versions of the phonological representations hypothesis were investigated by using a picture naming task and a battery of phonological measures at three linguistic levels (syllable, onset-rime, phoneme). The picture naming task was used to identify words for which dyslexic and control children had accurate vs inaccurate phonological representations, and performance in the phonological awareness tasks was then compared for the words which had precise vs imprecise representations. Findings indicated that frequency effects in the phonological awareness tasks at all levels disappeared for dyslexic and control children once representational quality was taken into account, and that the availability of sublexical units for analysis appeared to differ according to (1) the accuracy and retrieval of the phonological representation and (2) the linguistic level tapped by the phonological awareness task. PMID- 9226933 TI - Children's perception of faces of varied immaturity. AB - This research examined the relationship between facial immaturity and the perception of youthfulness, helplessness, and cuteness. In the first study, college students rated 16 faces for youthfulness. Faces varied within four dimensions (eye position, eye size, nose length, and shape of chin) representing either a mature or immature feature. College students rated faces conserving immature features as more youthful than those without those features. In the second study, three groups of children (5 to 8, 9 to 12, and 13 to 16 years old) rated the same 16 faces with respect to cuteness, helplessness, and youthfulness. Children were similar with respect to their attention to immature features when evaluating faces for youthful qualities, although older children were more sensitive to eye position than younger children when rating faces for youthfulness and helplessness. Older children were more consistent in their attention to immature features when rating faces. PMID- 9226934 TI - Emergent conditional discriminations in children and adults: stimulus equivalence derived from simple discriminations. AB - This study examines whether trained and derived simple discriminations lead to conditional relations between discriminative stimuli of the same and opposite (S+, S-) functions. After being trained on an arbitrary X-Y task (X1-Y1, X2-Y2) and on two simple discrimination tasks (A1+/A2- and B1+/B2-), children (Experiment 1) and adults (Experiment 2) were tested on the formation of novel simple discriminations (A3+/A2- and B3+/B2-) and conditional stimulus relations between all directly and indirectly paired A stimuli and between all directly and indirectly paired B stimuli (A1-A2-A3, B1-B2-B3). Subjects who formed these sets also received A2-X1 and B1-X2 training followed by a series of probes to assess the formation of two five-member stimulus equivalence classes (A1-A2-A3-X1-Y1, B1 B2-B3-X2-Y2). A modest majority of the children matched the directly paired stimuli (A2-A1, B2-B1 and A1-A2, B1-B2; A2-A3, B2-B3 and A3-A2, B3-B2) with one another while only a few of them also matched the indirectly paired stimuli with one another (A1-A3, B1-B3 and A3-A1, B3-B1). Those who did also related all the A and B stimuli with the designated X and Y stimuli. By contrast, all normal adults matched all paired and conditionally linked stimuli with one another. Present findings and those of related studies on stimulus equivalence are discussed from a stimulus contiguity perspective. PMID- 9226935 TI - An exploration of why preschoolers perform differently than do adults in audiovisual speech perception tasks. AB - Preschoolers' perception of audiovisual speech is considerably less influenced by visual information than adults'. We test the hypothesis that experience correctly producing consonants plays a role in developing the underlying representation which mediates the perception of visible speech. We divided preschoolers into two groups: those who made substitution errors and those who did not. Using a newly developed methodology, we tested substituters, nonsubstituters, and adults in an auditory-only condition, a visual-only condition, and an audiovisual condition. There were no differences among groups in the auditory-only condition. Overall, children still showed less visual influence than adults. Among the children, substituters were poorer at lip-reading in the visual-only condition and showed less visual influence on the incongruent audiovisual tokens than did nonsubstituters. These results support our hypothesis that experience correctly producing consonants plays a role in the elaboration of the underlying representation. PMID- 9226936 TI - Preschool children's attention to television: visual attention and probe response times. AB - Moment-to-moment variations in the engagement of young children's cognitive capacity by televised material were examined using a secondary task paradigm. Thirty-five 5-year-olds watched a 35-min Sesame Street program containing three types of segments: normal segments, segments with scenes reordered, and segments with incomprehensible language audio tracks. While watching the program, children were to respond quickly to auditory probes distributed across all types of segments and positions within segments. Probe response times and visual attention were recorded. Major findings were: As indicated by longer probe response times, capacity was more effectively engaged if language was comprehensible, provided children were looking at television when probes were presented. If not looking, response times were equally fast across segment types. For normal segments only, there were increases in the engagement of cognitive capacity if a look at the television or the program content had been continuous for some time. The findings provide evidence for, but important refinements of, the hypothesis that young children's ongoing comprehension is a major determinant of their attention to television. PMID- 9226937 TI - Imaging 'intact' myofibrils with a near-field scanning optical microscope. AB - Fluorescently labelled myofibrils were imaged in physiological salt solution by near-field scanning optical microscopy and shear-force microscopy. These myofibrils were imaged in vitro, naturally adhering to glass while retaining their ability to contract. The Z-line protein structure of the myofibrils was antibody labelled and easily identified in the near-field fluorescence images. The distinctive protein banding structure of the myofibril was also seen clearly in the shear-force images without any labelling requirement. With the microscope in the transmission mode, resolution of the fluorescence images was degraded significantly by excessive specimen thickness (> 1 micron), whereas the shear force images were less affected by specimen thickness and more affected by poor adherence to the substrate. Although the exact mechanism generating contrast in the shear-force images is still unknown, shear-force imaging appears to be a promising new imaging modality. PMID- 9226938 TI - Evaluation of fracture planes and cell morphology in complementary fractures of cultured cells in the frozen-hydrated state by field-emission secondary electron microscopy: feasibility for ion localization and fluorescence imaging studies. AB - We have employed field-emission secondary electron microscopy (FESEM) for morphological evaluation of freeze-fractured frozen-hydrated renal epithelial LLC PK1 cells prepared with our simple cryogenic sandwich-fracture method that does not require any high-vacuum freeze-fracture instrumentation (Chandra et al. (1986) J. Microsc. 144. 15-37). The cells fractured on the substrate side of the sandwich were matched one-to-one with their corresponding complementary fractured faces on the other side of the sandwich. The FESEM analysis of the frozen hydrated cells revealed three types of fracture: (i) apical membrane fracture that produces groups of cells together on the substrate fractured at the ectoplasmic face of the plasma membrane; (ii) basal membrane fracture that produces basal plasma membrane-halves on the substrate; and (iii) cross-fracture that passes randomly through the cells. The ectoplasmic face (E-face) and protoplasmic face (P-face) of the membrane were recognized based on the density of intramembranous particles. Feasibility of fractured cells was shown for intracellular ion localization with ion microscopy, and fluorescence imaging with laser scanning confocal microscopy. Ion microscopy imaging of freeze-dried cells fractured at the apical membrane revealed well-preserved intracellular ionic composition of even the most diffusible ions (total concentrations of K+, Na+ and Ca2+). Structurally damaged cells revealed lower K+ and higher Na+ and Ca2+ contents than in well-preserved cells. Frozen-freeze-dried cells also allowed imaging of fluorescently labelled mitochondria with a laser scanning confocal microscope. Since these cells are prepared without washing away the nutrient medium or using any chemical pretreatment to affect their native chemical and structural makeup, the characterization of fracture faces introduces ideal sample types for chemical and morphological studies with ion and electron microscopes and other techniques such as laser scanning confocal microscopy, atomic force microscopy and near-field scanning optical microscopy. PMID- 9226939 TI - Three-dimensional reconstruction of senile plaques in Alzheimer's disease. AB - So far, the three-dimensional approach to senile plaques, one of the principal histopathological hallmarks in Alzheimer's disease besides neurofibrillary tangles, has been scarce. Two main problems in three-dimensional reconstruction of histological specimens are the horizontal distortion during the preparation of serial thin tissue-slides and the need for consecutive vertical readjustment. This is greatly facilitated by the reflection contrast microscope (Leica, Germany) which is a light microscopical instrument causing interference patterns and reflections along interfaces by means of circularly polarized epi illumination. Using this technique, one can obtain distinct optical sections of a depth of 1.5 microns within specimens up to 30 microns in thickness, thus preserving the integrity of the observed object and rendering a manual alignment superfluous. We applied the reflection contrast microscope (RCM) on thick tissue slides of the cerebral cortex of a patient suffering from Alzheimer's disease which had been dyed according to Campbell. This is a silver-based staining method detecting beta A4-amyloid, the main component of senile plaques. Under the RCM, these silver-stained extracellular amyloid deposits cause reflections which allow the assessment of their three-dimensional distribution by focusing through the specimen. The optical sections obtained in this way were digitized, and the identified senile plaques reconstructed by the grey-scale image analysis system VIDAS 2.5 (Zeiss/Kontron, Germany). PMID- 9226941 TI - Evidence for change in children's use of defense mechanisms. AB - Using a cohort-longitudinal design, children's use of the defenses of denial, projection, and identification was assessed at four points over a 2-year time span, covering the age period from 6 years, 6 months to 9 years, 5 months. The results showed a clear decrease in the use of denial from early to middle childhood, a sharp increase in the use of projection, and an increase in the use of identification. These findings, predicted by a theory of defense mechanism development, are consistent with cross-sectional results reported previously. PMID- 9226940 TI - Plaster or plasticity: are adult work experiences associated with personality change in women? AB - The present study tested whether work experiences were associated with personality change across two periods of adulthood (age 21 to 27 and 27 to 43) in a longitudinal sample of women (N = 81). Two competing theoretical perspectives were tested: the plaster theory, which claims that personality does not change after age 30, and the plasticity theory, which claims that personality can change at any time in adulthood. Evidence was found for both correlational consistency of personality in adulthood and for the socialization effect of work on personality change. Work experiences were not associated with personality change in young adulthood but were associated with changes between young adulthood and midlife. In the period from age 27 to age 43 women who worked more became more agentic, and women who were more successful in their work became both more agentic and more normadhering. This pattern of associations between personality change and work experience provided support for the plasticity model of personality change. PMID- 9226942 TI - Adult attachment styles, the desire to have children, and working models of parenthood. AB - College students who had yet to marry and begin a family were asked about their desire to have children and their beliefs and expectations about themselves as parents (Study 1) and the characteristics of their prospective children (Study 2). Persons with more avoidant and anxious-ambivalent models of close adult relationships harbored more negative models of parenthood and parent-child relationships. These findings indicate that working models of parenting and parent-child relationships form well before marriage and the birth of children and that these models are systematically associated with attachment styles in adult relationships. The findings also suggest ways in which insecure attachments between child and parent may be influenced by the caregiver's models of parenting and parent-child relationships. PMID- 9226943 TI - Implications of mothers' personality for their parenting and their young children's developmental outcomes. AB - Using a recent model (Watson, Clark, & Harkness, 1994), we examined implications of mothers' personality (N = 103) for parenting and children's developmental outcomes, using multiple personality self-reports, lengthy, repeated naturalistic observations, and mothers' reports about parenting and their child. Mothers high in negative emotionality and disagreeableness showed more negative affect and their children were more defiant and angry; they also reported more power assertive and less nurturant parenting, as well as less secure attachment, more behavioral problems, and lower internalization of rules in their children. Mothers high in constraint and California Psychological Inventory (CPI) socialization reported more secure attachment and better internalization of rules; CPI socialization also correlated negatively with observed maternal verbal power assertion and children's defiance and anger, and positively with compliance. Regression analyses indicated that mothers' personality, particularly negative emotionality and socialization, influenced broadly conceptualized adaptive child outcomes, even after the influence of parenting was controlled. PMID- 9226944 TI - Stability and change in psychosocial resources during caregiving and bereavement in partners of men with AIDS. AB - This study examines the effects of caregiving and bereavement on psychosocial resources in HIV+ and HIV- caregivers of men with AIDS. We explored three hypotheses regarding these effects: the "wear and tear" hypothesis, which asserts that the chronic stress of caregiving and bereavement diminishes resources; the "enhancement" hypothesis, which asserts that caregiver resources may increase in response to increased demands; and the "personality" hypothesis, which asserts that psychosocial resources reflect stable personality characteristics. We addressed four questions: (a) What are the effects of caregiving on resources? (b) How do these resources vary by the imminence of the partner's death? (c) What is the effect of the partner's death on these resources? and (d) How does the caregivers' HIV serostatus influence the effects of caregiving and bereavement on resources? Support for the personality hypothesis predominated, with some support for the wear and tear hypothesis, depending on the resource in question. In general, HIV seropositivity did not put people at additional risk for resource depletion. PMID- 9226945 TI - Micro- and macrogeographical genetic structure of colonies of naked mole-rats Heterocephalus glaber. AB - Patterns of genetic structure in eusocial naked mole-rat populations were quantified within and among geographically distant populations using multilocus DNA fingerprinting and mitochondrial DNA (mtDNA) sequence analysis. Individuals within colonies were genetically almost monomorphic, sharing the same mtDNA control region haplotype and having coefficients of band sharing estimated from DNA fingerprints ranging from 0.93 to 0.99. Family analysis of a hybrid captive colony of naked mole-rats with increased levels of genetic variability using multilocus DNA fingerprinting gave results consistent with Mendelian inheritance, and has revealed for the first time that multiple paternity can occur. In a survey of wild colonies from Ethiopia, Somalia and locations in northern and southern Kenya, we have examined mtDNA control region sequence variation in 42 individuals from 15 colonies, and together with multilocus DNA fingerprinting and mtDNA cytochrome-b sequence analysis in selected individuals have shown that these populations show considerable genetic divergence. Most of the variance in sequence divergence was found to be between geographical locations (phi ct = 0.68) and there was a significant correlation between sequence divergence and geographical separation of haplotypes. Six colonies from Mtito Andei in southern Kenya shared the same control region haplotype, suggesting a recent common maternal ancestor. In contrast, out of four colonies at Lerata in north Kenya, three haplotypes were identified, and phylogenetic analysis suggests that this area may be a zone where two distinct lineages are in close proximity. Genetic distances were maximal between Ethiopian and southern Kenyan populations at 5.8% for cytochrome-b, and are approaching interspecific values seen between other Bathyergids. PMID- 9226946 TI - Molecular evidence for an extinct parent of the tetraploid species Microseris acuminata and M. campestris (Asteraceae, Lactuceae). AB - To determine the origin of the tetraploid annuals Microseris campestris and M. acuminata, chloroplast RFLP, RAPD and ITS sequence variability among nine populations of the two polyploids and 14 populations of the diploid annuals M. elegans and M. douglasii have been surveyed. Previously described variable chloroplast restriction sites infer M. douglasii as the possible maternal parent of both tetraploid species. However, the chloroplast genome typical for M. douglasii has now also been found in some plants of M. elegans. RAPD analysis revealed 172 polymorphic DNA markers that defined all four species as genetically distinct groups, but demonstrated closer associations between M. douglasii and M. acuminata, and between M. elegans and M. campestris. Sequencing of the ITS-1 and ITS-2 region yielded 73 phylogenetically informative sites. Thirty basepair mutations separated the annual Microseris species from the outgroup, Uropappus lindleyi. The putative interspecific allotetraploid M. campestris contained only one type ITS sequence that, on the basis of eight synapomorphic substitutions was derived from M. elegans. The single ITS of M. acuminata shares six common sites with M. douglasii. Surprisingly, six sites were synapomorphic for the two tetraploids, M. campestris and M. acuminata, suggesting recombination within the ITS of both species with that of a common, now extinct, parental taxon, possibly the donor of the M. douglasii type chloroplasts found in both tetraploids. These results confirm the interpretation of M. campestris as derived from M. douglasii (extinct population) and M. elegans, and resolve the unknown origin of M. acuminata as an intraspecific hybrid between two very distinct populations of M. douglasii, one of them the same extinct M. douglasii form that contributes to M. campestris. PMID- 9226947 TI - Interspecific microsatellite markers for the study of pinniped populations. AB - Microsatellites have rapidly become the marker of choice for a wide variety of population genetic studies. Here we describe 20 pinniped microsatellite markers which have been tested across 18 pinniped species. The majority of these markers have broad utility in all pinnipeds and provide a strong base for detailed population genetic studies in the Pinnipedia. PMID- 9226948 TI - An effective method for isolating DNA from historical specimens of baleen. AB - DNA was isolated from an early twentieth century museum specimen of northern right whale baleen. A system of stringent controls and a novel set of cetacean specific primers eliminated contamination from external sources and ensured the authenticity of the results. Sequence analysis revealed that there were informative nucleotide positions between the museum specimen and extant members of the population and closely related species. The results indicate that museum specimens of baleen can be used to assess historical genetic population structure of the great whales. PMID- 9226949 TI - A RNA/RNA heteroduplex cleavage analysis to detect rare mutations in populations. PMID- 9226950 TI - Microsatellites in the bottlenose dolphin Tursiops truncatus. PMID- 9226952 TI - Toxoplasmosis in rats (Rattus norvegicus): congenital transmission to first and second generation offspring and isolation of Toxoplasma gondii from seronegative rats. AB - To study congenital transmission of Toxoplasma gondii during acute and chronic infections, 4 pregnant Sprague-Dawley rats were each fed 10,000 oocysts of the VEG strain. Toxoplasma gondii was recovered from 33, 55, 83 and 57% of rats (F1) when dams were inoculated at 6, 9, 12 or 15 days of gestation, respectively. Progeny of 15 congenitally infected female rats were examined for T. gondii. Toxoplasma gondii was recovered from tissues of 1 of 155 rats (F2) born to congenitally infected dams. A total of 4 (F2) females were mated; 0 of 40 (F3) rats born to them were infected. None of the acutely infected 4 dams that had given birth to congenitally infected litters produced congenitally infected offspring during the second pregnancy. Thus, unlike mice, evidence for repeated congenital transmission of T. gondii in the rat was found in < 1% of cases. Of the 16 congenitally T. gondii infected pups with demonstrable tissue cysts, 5 were seronegative (< 1:4) in the Sabin-Feldman dye test and 5 were seronegative (< 1:20) in the modified agglutination test by the use of whole formalinized tachyzoites and mercaptoethanol. PMID- 9226951 TI - Characterization of microsatellite loci in caribou Rangifer tarandus, and their use in other artiodactyls. PMID- 9226953 TI - Tissue cyst tropism in Toxoplasma gondii: a comparison of tissue cyst formation in organs of cats, and rodents fed oocysts. AB - The persistence of Toxoplasma gondii tissue cysts in organs of cats (definitive host) and rodents (intermediate hosts) was studied. Nine cats, 12 rats, and 12 mice were fed T. gondii oocysts and their organs were digested in pepsin and then bioassayed for bradyzoites in mice. Of 9 cats killed 37 or 51 days after feeding 10(2) (2 cats), 10(3) (3 cats) or 10(4) (4 cats) oocysts of the VEG strain, tissue cysts were found in each cat; in the tongue of 9, in the heart of 5, in the brain of 4, and in the eyes of 1 cat. The dose had no effect on the distribution of tissue cysts in cats. Twelve rats were each fed 10(5) oocysts of the VEG strain of T. gondii and killed 21, 29, 64 or 237 days later. At each time period, 11 tissues of 3 rats were pooled and bioassayed in mice. Tissue cysts were found in the brain, skeletal muscle, heart and kidneys of rats at each killing time; in the lungs, intestines, and mesenteric lymph nodes in 3 of 4 instances; in the tongue, liver, and eyes in 2 instances and in the spleen in 1 instance. Also, using the same procedures and sampling the same 11 tissues as used for rats, tissue cysts were seen in all organs except in the tongue and liver of 3 mice killed on day 82 after feeding the VEG strain. In 9 mice (3 with each strain) fed oocysts of the ME-49, GT-1, or P89 T. gondii strain and killed 62-130 days later, tissue cysts were found consistently only in the brain. Thus, in rats and mice, most tissue cysts were found in the brain and rarely in the tongue. This was in marked contrast to the distribution of tissue cysts in cats. PMID- 9226955 TI - Continuous in vitro culture of Babesia divergens in a serum-free medium. AB - Babesia divergens was cultivated in RPMI 1640 (25 mM HEPES) supplemented with 10% human serum (RPMI-10% HS) with a high percentage of parasitized erythrocytes (PPE) (> or = 40%). Standardization of in vitro tests, purification of exoantigens, biochemical studies and the safety of the culture handler motivated the development of a serum-free defined medium. Removal of serum greatly reduced the PPE but, after a period of adaptation, the culture was continuous and the parasite was able to develop a 3% routine PPE. Addition of vitamins or reduced glutathione in basal medium (RPMI) did not improve the PPE. The supplementation of basal medium with lipidic carrier (Albumax I or bovine serum albumin-Cohn's fraction V) promoted the growth of B. divergens with high PPE (> 30%) close to those obtained in RPMI-10% HS. Neither protein nor lipid fractions alone were able to restore the growth of B. divergens. Nevertheless, the whole lipid fraction from serum or Albumax I added to delipidated albumin partially restored the growth (7% PPE), indicating that the presentation of specific lipids by a carrier is crucial for the parasite. All the data indicate that Albumax I can replace human serum offering the advantages of safety, standardization for chemosensitivity tests, and exoantigen purification. PMID- 9226954 TI - A method for the isolation of schistosome eggs and miracidia free of contaminating host tissues. AB - A novel method for the isolation of schistosome eggs and miracidia from livers of mice infected with Schistosoma japonicum or S. mansoni is described. The method employed collagenase B to degrade the interstitial matrix of mouse liver tissue, after which the schistosome eggs were separated from the liver cells by 2 single step density centrifugations through Percoll. Using this procedure sufficient quantities of miracidia were obtained to generate a cDNA library. Southern blot analysis demonstrated that miracidia isolated by this method were free from contaminating host DNA. PMID- 9226956 TI - Identification of an Ixodes ricinus salivary gland fraction through its ability to stimulate CD4 T cells present in BALB/c mice lymph nodes draining the tick fixation site. AB - BALB/c mice infested with larvae or nymphs of Ixodes ricinus develop in their lymph nodes a T cell-specific immune response triggered by salivary gland soluble antigens (SGA). SGA are apparently conserved in the 3 biological stages of I. ricinus ticks and are species specific. SGA derived from partially fed females I. ricinus stimulate lymph node T cells from mice infested with I. ricinus larvae or nymphs. In contrast, lymph node cells from mice infested with Amblyomma hebraeum nymphs do not respond. A chromatographic fraction enriched with a 65 kDa protein (IrSG65) isolated from salivary glands of I. ricinus partially fed females induces in vitro a specific T cell proliferation of lymph node cells from mice infested with I. ricinus nymphs. The depletion of CD4+ T cells drastically reduces the ability of lymphocytes from infested mice to proliferate after IrSG65 stimulation. PMID- 9226957 TI - Cloning of peroxiredoxin, a novel antioxidant enzyme, from the helminth parasite Fasciola hepatica. AB - A cDNA was isolated from a cDNA expression library using serum prepared against a high molecular mass fraction of Fasciola hepatica excretory-secretory products. The full-length cDNA encodes a member of the recently described peroxiredoxin antioxidant family. Peroxiredoxin could be the major hydrogen peroxide removing antioxidant in F. hepatica since this parasite does not express a catalase and expresses little glutathione peroxidase activity. This novel antioxidant may be involved in functions such as protection against reactive oxygen species (ROS) generated by metabolic processes and/or protection of the parasite against ROS released by immune effector cells. PMID- 9226958 TI - Impairment of the chemical defence of the beetle, Tenebrio molitor, by metacestodes (cysticeroids) of the tapeworm, Hymenolepis diminuta. AB - The defensive glands of beetles, Tenebrio molitor, infected with metacestodes (cysticercoids) of Hymenolepis diminuta are everted less frequently upon stimulation, and contain less toluquinone (methylbenzoquinone) and m-cresol, than glands of uninfected controls. These differences, as shown in predation trials with wild rats, increase the likelihood that both cysticercoids and beetles will be ingested by the tapeworm's definitive host. This is the first documented case of a parasite inhibiting the chemical defence of an intermediate host, and one of only a few reports of parasite-induced manipulation of host biology supported by empirical evidence implicating facilitated parasite transmission between host species. PMID- 9226959 TI - Enhancement of recombinant glucoamylase expression by introducing yeast GAL7 mRNA termination sequence. AB - Glucoamylase gene (STA1) of Saccharomyces diastaticus was expressed in recombinant Saccharomyces cerevisiae systems. The yeast, GAL7 mRNA termination sequence, was introduced in the 3' noncoding region of the STA1 structural gene which was under the control of the SUC2 promoter and STA1 secretion signal sequence. This plasmid was named YEpSSG7 and was introduced into yeast S. cerevisiae MMY2 to construct recombinant S. cerevisiae MMY2SSG7. The GAL7 mRNA termination sequence enhanced the glucoamylase expression level by 3-5 times depending on the culture conditions compared to the result from the strain S. cerevisiae MMY2SUCSTA which did not contain the GAL7 mRNA termination sequence. Such an enhancement was not due to plasmid stability or plasmid copy number effects. Such an enhancement was primarily due to the fact that GAL7 mRNA termination sequence stabilized the STA1 mRNA 3' end. PMID- 9226960 TI - Plasmid instability kinetics in continuous culture of a recombinant Saccharomyces cerevisiae in airlift bioreactor. AB - Plasmid instability of a recombinant Saccharomyces cerevisiae C468/pGAC9 (ATCC 20690) was examined during continuous culture in a nonselective medium in an airlift bioreactor. The recombinant strain contained a 2-micron based shuttle vector pGAC9 and expresses Aspergillus awatnori glucoamylase gene under the control of the yeast enolase I (ENO1) promoter. The changes in the fraction of plasmid-bearing cells and glucoamylase activity followed first-order kinetics. Expressed as a function of time, the decay rates of both the plasmid-bearing cell fraction and glucoamylase expression increased with increasing dilution rates. If expressed as a function of cell generations, the decay rates were nearly constant over the dilution rates tested. The results indicated that the growth rate difference between plasmid-bearing and plasmid-free cells was negligible. This was probably due to the low copy number of the 2-micron based yeast shuttle vector. Thus the contribution of preferential growth to apparent plasmid instability was negligible. A novel numerical method is proposed to evaluate the parameters related to plasmid stability. The estimated values of probability of plasmid loss (P = 0.0499) were nearly constant at different dilution rates. No significant effect of growth rates on plasmid instability was observed. The proposed kinetics agreed well with experimental observations. PMID- 9226961 TI - Over-expression of the Saccharomyces cerevisiae exo-beta-1,3-glucanase gene together with the Bacillus subtilis endo-beta-1,3-1,4-glucanase gene and the Butyrivibrio fibrisolvens endo-beta-1,4-glucanase gene in yeast. AB - The EXG1 gene encoding the main Saccharomyces cerevisiae exo-beta-1,3-glucanase was cloned and over-expressed in yeast. The Bacillus subtilis endo-1,3-1,4-beta glucanase gene (beg1) and the Butyrivibrio fibrisolvens endo-beta-1,4-glucanase gene (end1) were fused to the secretion signal sequence of the yeast mating pheromone alpha-factor (MF alpha 1S) and inserted between the yeast alcohol dehydrogenase II gene promoter (ADH2P) and terminator (ADH2T). Constructs ADH2P MF alpha 1S-beg1-ADH2T and ADH2P-MF alpha 1S-end 1-ADH2T designated BEG1 and END1, respectively, were expressed separately and jointly with EXG1 in S. cerevisiae. The construction of fur 1 ura3 S. cerevisiae strains allowed for the autoselection of these multicopy URA3-based plasmids in rich medium. Enzyme assays confirmed that co-expression of EXG1, BEG1 and END1 enhanced glucan degradation by S. cerevisiae. PMID- 9226962 TI - Polymorphic microsatellite DNA markers in the rabbit (Oryctolagus cuniculus). AB - By searching the EMBL nucleotide database a total of 157 rabbit nuclear gene microsatellites were obtained (VAN LITH and VAN ZUTPHEN, Animal Genetics 27, 387 395, 1996). Thirteen of these were analysed by PCR to examine the degree of polymorphism of the amplified fragments in rabbits from different breeds. The 13 pairs of primers resulted in polymorphic products with an average of four alleles per microsatellite sequence (ranging between 2-11). There was a positive relationship between the longest repeat unit number in the nucleotide sequence and the number of alleles detected. The results obtained so far justify the conclusion that rabbit microsatellites extracted from the EMBL nucleotide sequence database are sufficiently polymorphic to be useful as Type 1 markers in rabbit genetic studies. PMID- 9226963 TI - Response of the germfree rat colonic mucosa to dietary endotoxins. AB - Suggestive to induce an immunoreactive response in the colon mucosa of germfree rats 3 diets, autoclaved or gamma-irradiated, respectively, were administered to groups of 30 days old GF Ztm:SPRD rats. Diets and rats were studied in parallel after a feeding period of 20 days. The microbiological controls confirmed the sterility of diets and animals. The aqueous suspensions of one diet displayed obviously dead yeast and Gram positive and negative bacteria, which consistently were evident in colonic cast preparations of animals fed with this diet. These findings apparently were not related to the pathohistological alterations, observed in hematoxilin-eosin stained colon sections. However, dietary endotoxin concentrations between 0.6 and 10 micrograms LPS/g corresponded with the endotoxin concentrations in feces (0.3 to 3.1 micrograms LPS/g wet weight) and in colonic tissue (0.01 to 0.6 microgram LPS/g wet weight). These endotoxins obviously mediated a dose-dependent immunoreactive response of the colonic mucosa: in parallel to the dietary endotoxin content, the cellular infiltrations ranged from single mononuclear cells to severe, cell mediated, mucosal alteration. GF rats, used as an experimental animal model for inflammatory disorders of the intestine obviously necessitate diets with low endotoxin concentrations. PMID- 9226964 TI - Time of day and stress response to different stressors in experimental animals. Part I: Golden hamster (Mesocricetus auratus Waterhouse, 1839). AB - The present paper describes the effects of animal house routine stressors on adult golden hamsters during activity time (2 hrs after lights off) and rest time (2 hrs after lights on). In addition, for determination of norm values, the circadian rhythms of the stress indicators heart rate, core body temperature and general activity of unstressed animals were telemetrically registered via implanted transmitters. The three circadian patterns of the nocturnal golden hamster under L:D = 12:12 were unimodal with a main peak after lights off. The physiological norm values (mean over 24 hours +/-SD) were: heart rate 324 +/- 18 bpm, core body temperature 37.5 +/- 0.5 degrees C and activity 114 +/- 123 units/5 min. The mean body temperature of females was significantly higher (0.4 degree C) and its mean activity level was significantly (40%) lower than that of males. The stress responses were dependent on the time of day and on the kind of stressor. The stress responses were significantly stronger during the rest time of the animals (i.e. light period), and it resulted in the subsequent ranking of stressors: handling < vaginal smear < intruder/resident confrontation < cage changing < grouping. There were no sex-dependent stress response differences. The results of this study were compared with identical investigations on the social Mongolian gerbil (J. Exp. Anim. Sci. 1996/97; 38: No. 3). PMID- 9226965 TI - A new technique: serial puncture of the cisterna magna for obtaining cerebrospinal fluid in the mouse--application in a model of herpes simplex virus encephalitis. AB - This report describes a new technique for obtaining cerebrospinal fluid from the living mouse (SJL/NBom) in a model of herpes simplex virus encephalitis which is also applicable to other mouse models. The puncture technique was performed in living animals which had been infected with Herpes Simplex Virus Type I strain F in the living animal. The cisterna magna was micro-surgically prepared: The neck muscles were dissected microscopically down to the dura which subsequently was punctured by a glass micropipette. This newly developed minimally invasive technique was performed in a group of living animals (n = 20) and results compared with those of a second group of perfusion fixed animals (n = 20). For the first time, repeated cerebrospinal fluid punctures of individual, living animals are possible. This is of great value for the assessment of new therapeutic and diagnostic strategies in experimental research using mouse models. In addition, this refined methodology significantly reduces the number of experimental animals. PMID- 9226966 TI - A technique for extracorporeal circulation in the Goettingen minipig allowing recovery and long-term follow-up. AB - The objective of our study was to establish an animal model with extracorporeal circulation (ECC) for investigations on the long-term tolerability of different pericardial substitutes using the Goettingen minipig. A combination of halothane (0.4-0.6% vaporizer setting) in oxygen and nitrous oxide (FiO2 0.33), with the opioid piritramide (75 micrograms/kg/h i.v.) was used for anaesthesia. A muscle relaxant was not administered. Due to the size of the animals (mean body weight 37.6 kg) the operative procedure was the same as in humans. Specific changes preceding or following initiation of cardiopulmonary bypass (CPB), such as inactivation of the coagulation system (400 IU heparin/kg i.v.), haemodilution, and hypothermia (32 degrees C), did not result in any complication. We did not induce cardiac arrest during CPB in order to facilitate haemodynamic stability after weaning from CPB. In three animals a temporary increase in blood pressure occurred after protamine (2.5-3.0 mg/kg i.v.) was given to reverse the heparinization. Within 60 minutes after the end of surgery all animals could be extubated when spontaneous breathing and cough reflex were present. The postoperative follow-up period of nine months was uneventful apart from one animal which developed a superficial wound in the thoracic scar area. We conclude that our technique for ECC is a safe method allowing recovery and long-term follow-up after cardiac surgery in a porcine animal model. PMID- 9226968 TI - Humoral immune responses in mice infected with gamma-irradiated third-stage larvae of Angiostrongylus cantonensis. AB - Humoral immune responses in C57BL/6 strain mice infected with 1.0 kGy gamma irradiated third-stage larvae (L3) and with non-gamma-irradiated L3 of Angiostrongylus cantonensis were assessed in this study. The young-adult worm (L5) antigen-specific antibodies in sera of both groups of mice elevated gradually and reached a peak in the third week after infection. The IgM antibodies of mice 1-week after infection with gamma-irradiated L3 and with non gamma-irradiated L3 recognized several L5 antigens of the same molecular weight. However, only partially identical profiles were observed in the reaction of L5 antigens with IgM and IgG antibodies from mice 3 weeks after infection with gamma irradiated L3 and with non-gamma-irradiated L3 respectively by western blot analysis. With regard to antibody-dependent cell-mediated cytotoxicity, antibodies from 3-week infected mice significantly increased the adherence of mouse eosinophils and neutrophils to L3 of A. cantonensis. The immunoresponses of splenic cells from mice in the third week of infection to sheep red blood cells were suppressed because the number of plaque forming cells obviously decreased more than the uninfected control. PMID- 9226967 TI - General anesthesia for ureteral measurements in the rabbit. AB - Literature search and in vitro studies on ureteral function in humans and rabbits have proven that the rabbit is a suitable animal model for the investigation of the effect of smooth muscle relaxing substances on the ureter. One of the main problems encountered was to find an appropriate anesthetic protocol for this animal model. Application of barbiturates as a monotherapy proved to be unsuitable to allow painfree preparation of the abdomen. Intravenous (iv) anesthesia consisting of ketamine-HCl/xylazine-HCl could not be considered due to interference with ureteral smooth muscle tone. Intravenous administration of ketamine-HCl induced immediate ureteral contractions with increased frequency of ureteral activity. Xylazine-hydrochloride, a mixed alpha 2-, alpha 1-adrenoceptor agonist inhibits the increase in synthesis of 3'5'-cAMP. Since the test substances used are phosphodiesterase-IV-inhibitors (rolipram and its two enantiomers), which increase 3'5'-cAMP, this type of anesthesia would interfere with the pharmacological effect to be investigated. General anesthesia using a combination of nitrous oxide (2 l/min) and oxygen (1 l/min) and a very small amount (2 mg/kg b.w.) of pentobarbital i.v. every 30 minutes, was found to be the most suitable form of anesthesia. It resulted in much more stable circulatory conditions, sufficient depth of anesthesia and the possibility to test muscle relaxing substances (PDE-IV-inhibitor) without any influence from anesthesia on their efficacy. PMID- 9226969 TI - Changes of hepatitis B markers among young adults in a hepatitis B virus endemic area: a follow up study on medical students. AB - To elucidate the hepatitis B virus (HBV) infection status of teenagers, to observe changes of the status of these young adults after long-term follow up, to clarify the HBV replication activity of young HBsAg carriers, and to understand the anti-HBs titer of those immune to HBV-infection, a comparison between two health examinations at a 5-or 6-year interval was conducted among 132 medical students. Their ages were 19.1 +/- 0.9 (mean +/- SD). There were 19 (14.4%) HBsAg carriers, 57 (43.2%) immune to HBV infection, and 56 (42.4%) susceptible to HBV. All 19 HBsAg positive carriers were still in carrier status in the second examination. Twelve (63.1%) of the HBV carriers had already HBeAg negative seroconversion before the age of 25. And two carriers might be in the late e seroconversion phase. One susceptible student had a subclinical HBV infection during his university life, with an incidence of HBV infection of 0.47% (1/213) per person-year. Nine (11%) of 82 HBV-immune students had lower anti-HBs concentrations. Six of the 9 students were immunized by natural infection. Anamnestic response developed after booster vaccination. This study shows that these young adults have an HBsAg-carrier rate similar to the general adult population in Taiwan. However, HBeAg seroconversion occurs more often and earlier in this particular population than previous reports have indicated. Moreover, only 24 (40%) susceptible subjects received HBV vaccination after the first recommendation and it is necessary to be more forceful in recommending prompt vaccination. PMID- 9226970 TI - Ambulatory 24-hour esophageal manometry and pH-metry in patients with noncardiac chest pain, but no reflux symptoms. AB - The aim of this study is to determine the diagnostic value of 24-hour ambulatory esophageal manometry and pH-metry for patients with noncardiac chest pain (NCCP), but no reflux symptoms. Twenty-four hour ambulatory esophageal manometry and pH metry was performed on 34 patients with NCCP, but no reflux symptoms. The pressure transducers were located 3, 8, and 13 cm above the lower esophageal sphincter (LES) and the pH probes were located 5 and 20 cm above the LES. An event marker was triggered by the patient for chest pain. Only 17 patients (50%) had at least one pain episode (total 81 episodes, range 1-19 episodes per person) during a 24-hour recording. Twenty-one chest pain episodes (26%) occurred during abnormal motility, whereas 4 episodes (5%) were associated with pH < 4, and 10 episodes (12%) had both abnormalities. The majority of chest pain episodes, 46 out of 81 events (57%), did not have any association with motility or pH abnormalities. Five of 7 patients (71%) with reflux-related chest pain and 8 of 11 patients (73%) with dysmotility-related chest pain had symptom association probability > 95%, indicating a significant association between chest pain and esophageal dysfunction. Our conclusion is that ambulatory esophageal manometry and pH-metry is a useful tool in the evaluation of NCCP, but only a few additional patients with reflux-related chest pain could be found in patients without reflux symptoms. PMID- 9226971 TI - A mixture of allogeneic dermal tissue and autologous microskin grafting of rabbit skin wound. AB - In a rabbit model, transplantation of largely expanded microskin grafts (expansion ratio 25 X) were studied. In Group I, the autologous microskin grafts were transplanted onto the full-thickness skin defect on the back of a rabbit and were overlaid with Biobrane. In Group II, the autologous microskin grafts were mixed with allogeneic microdermal substance (expansion ratio 25 X) before the transplantation. The percentage of re-epithelialized area to the total grafted wound was analyzed by means of computerized planimetric analyses of photographs of the grafted wounds on days 11 and 13. Biopsies of the grafted wounds were done. On day 11, the mean percentage of Group I was 72.4% and that of Group II was 82.7%. On day 13, they were 82.3% in Group I and 92.3% in Group II. In Group II, the allogeneic dermal tissue did not cause obvious rejection in the neoepithelium. Histological features showed the existence of allogeneic dermal tissue in the grafted wound. The adding of largely expanded allogeneic dermal substance to autologous microskin grafts in Group II provided better circumstances than that in Group I for re-epithelialization of autologous mciroskin grafts. PMID- 9226972 TI - Two-dimensional electrophoretic analyses of urinary proteins in Chinese male urinary stone patients. AB - This study was aimed to detect the differences in soluble urinary proteins between normal Chinese male individuals and urinary stone formers by 2 dimensional electrophoresis. Twenty urine samples were obtained from normal male adults and 35 from recurrent male urinary stone formers. The stone compositions included calcium oxalate, uric acid, carbonate apatite, and brushite. Two hundred milliliters of fresh urine was collected for analysis. Each urine sample was concentrated, dialyzed, frozen and lyophilized. The samples of the same stone composition were pooled and subjected to two-dimensional electrophoresis. Comparison of protein profiles between normal individuals and stone formers revealed a number of proteins which are not present in the urine of normal individuals, 5 from calcium oxalate, 2 from uric acid, 3 from carbonate apatite and 2 from brushite. These urinary stone-associated proteins comprise proteins A (37kd), B (30kd), C (26kd), D (25kd), and E (22kd) for calcium oxalate, proteins F (63kd) and G (59kd) for uric acid, protein H (65kd), I (42kd), and J (30kd) for carbonate apatite as well as proteins K (61kd) and L (59kd) for brushite. Among them, the proteins A, I, and J exhibited charge heterogeneity. PMID- 9226973 TI - Prevalence of human papilloma virus 16 or 18 in cervical cancer in Hualien, eastern Taiwan. AB - In order to determine the association of human papilloma virus (HPV) with cervical cancer of patients in the Hualien area, we analyzed 40 cervical cancer specimens using polymerase chain reaction (PCR) to detect the presence of HPV type 16 (HPV-16) and type 18 (HPV-18) genomes. The results showed that at least 70% (28/40) of the specimens had HPV DNA. Of the 37 squamous cell carcinomas of the cervix, HPV-16 was present in 25 cases (68%) and HPV-18 in 2 (5.4%). HPV-16 DNA was detected in one of 2 adenocarcinomas and HPV-18 in a case of small cell carcinoma. Seven (87.5%) of 8 specimens from aborigines were HPV-positive. These findings support a role for HPV in the development of cervical cancer in the Hualien population. PMID- 9226974 TI - Myofibroblastoma of the breast: a case report. AB - A tumor in the breast of a 67-year-old male was resected. It was grossly circumscribed and unencapsulated. Histologically, it was composed of plump and long bipolar, spindle cells arranged in fascicular clusters with intervening broad collagen bands. The cells lacked pleomorphism, and no mitotic figures were found. The margin was that of pushing border type. No epithelial component was seen in the tumor. Immunohistochemically, the tumor cells expressed variable reactivity to vimentin, desmin and smooth muscle actin and were negative for S 100 protein and keratin. Electron microscopy showed cell differentiation between fibroblast and smooth muscle cells. PMID- 9226975 TI - Gaze-evoked amaurosis caused by intraconal cavernous hemangioma: a case report. AB - Gaze-evoked amaurosis is an uncommon but pathognomonic symptom of intrinsic orbital mass lesion. The symptom is easily neglected even by ophthalmologists. A 49-year-old woman had a two-year history of dimming vision in her right eye when she looked upward for a few seconds. On examination, her visual acuity was normal and without apparent proptosis. The orbital CT scan and MR imaging showed an intraconal cavernous hemangioma just adjacent to the optic nerve in the right orbit. The pattern visual-evoked potential (PVEP) on primary position was normal in both eyes, but a latency delayed (P100) in the right eye was found after she looked upward for a few seconds. We discuss this interesting finding and review the possible mechanisms involved. PMID- 9226976 TI - Bacterial meningitis and lumbar epidural hematoma due to lumbar acupunctures: a case report. AB - A 48-year-old female expressed signs of meningeal irritation after having received several lumbar acupunctures within one week for back pain. Bacterial meningitis was diagnosed from cerebrospinal fluid examinations. Magnetic resonance image (MRI) of spine at admission demonstrated a fusiform lesion with characters of subacute hematoma in the epidural space of the first and second lumbar level. She received antibiotics treatment only and recovered from her central nervous system infection completely. The epidural lesion disappeared spontaneously in the MRI follow up three weeks later. We report the diagnosis and follow-up of epidural hematoma of the lumbar spine by MRI which aided the medical physician to treat meningitis attentively. PMID- 9226977 TI - The problem of quality of life. PMID- 9226978 TI - Is the SF-36 health survey questionnaire suitable as a self-report measure of the health status of older adults with Parkinson's disease? AB - An amended version of the Short Form 36 health survey questionnaire (SF-36), which has been suggested as being more suitable for self-completion in older adults, was evaluated among a group of elderly subjects with Parkinson's disease (PD). In the subjects who were able to fully complete the modified SF-36 unaided, the severity of PD was broadly reflected by low reported levels of functioning and well-being with this measure. However, the modified questionnaire still did not overcome the problem of missing responses from elderly subjects. A total of 24% of respondents missed one or more of the 36 statements and 80% of missing responses were concentrated in the three statements which had been modified. The SF-36 in its original form would need to be combined with a disease-specific measure to adequately evaluate the health status of older adults with PD. PMID- 9226979 TI - Assessing the validity of the SF-36 General Health Survey. AB - Our objective was to assess the validity of the SF-36 General Health Survey against the Social Maladjustment Schedule (SMS) and two questionnaire measures, the Social Problem Questionnaire and the Nottingham Health Profile (NHP) in a random subsample of 206 men and women from the Whitehall II study, a longitudinal survey of health and disease amongst 10,308 London-based civil servants. We found that social functioning on the SF-36 correlated significantly with social contacts, total satisfaction and total management scores on the SMS, and social isolation and emotional reactions on the NHP. General mental health on the SF-36 was associated with marriage, social contacts, leisure scores, total satisfaction and total management scores on the SMS, and emotional reactions, energy level and social isolation on the NHP. Conversely, physical functioning and physical role limitations were generally not associated with the SMS but were associated with physical abilities and pain on the NHP. In conclusion, this study offers evidence of the discriminant validity of the general mental health and physical functioning scales of the SF-36. We also found moderate construct and criterion validity for the social functioning scale of the SF-36 and considerable overlap between the general mental health and social functioning scales. PMID- 9226980 TI - Randomized trials with quality of life endpoints: are doctors' ratings of patients' physical symptoms interchangeable with patients' self-ratings? AB - The assessment of physical symptoms is a key component of quality of life studies in palliative care, but is often hampered by missing data from patient-completed questionnaires. In two large multicentre randomized trials of palliative treatment conducted by the Medical Research Council Lung Cancer Working Party, Involving over 700 patients, patients completed Rotterdam Symptom Checklists and doctors reported on eleven of the same physical symptoms at each assessment, using the same 4-point severity scale. Ratings by doctors and patients were compared with respect to compliance, severity, and outcomes for the respective trials. Doctors provided more data than patients: 66% vs. 52% in the first 6 months in one trial, 58% vs. 61% in the other. Comparisons of over 33,000 symptom assessments showed 78% complete agreement between doctor and patient, 18% disagreement by one, 4% two, and 1% three grades (complete disagreement). There was no change in levels of agreement over time, but increasing disagreement with increasing symptom severity, and a consistent bias towards doctors underestimating severity. Nevertheless, the two methods of data collection resulted in similar between-treatment conclusions. Therefore, in randomized trials the doctors' assessments of key physical symptoms may be sufficient for the between-treatment comparison. However, the fact that doctors underestimate symptom severity 15% of the time has important implications for palliative interventions. PMID- 9226981 TI - 'Equivalence' and the translation and adaptation of health-related quality of life questionnaires. AB - The increasing use of health-related quality of life (HRQOL) questionnaires in multinational studies has resulted in the translation of many existing measures. Guidelines for translation have been published, and there has been some discussion of how to achieve and assess equivalence between source and target questionnaires. Our reading in this area had led us, however, to the conclusion that different types of equivalence were not clearly defined, and that a theoretical framework for equivalence was lacking. To confirm this we reviewed definitions of equivalence in the HRQOL literature on the use of generic questionnaires in multicultural settings. The literature review revealed: definitions of 19 different types of equivalence; vague or conflicting definitions, particularly in the case of conceptual equivalence; and the use of many redundant terms. We discuss these findings in the light of a framework adapted from cross-cultural psychology for describing three different orientations to cross-cultural research: absolutism, universalism and relativism. We suggest that the HRQOL field has generally adopted an absolutist approach and that this may account for some of the confusion in this area. We conclude by suggesting that there is an urgent need for a standardized terminology within the HRQOL field, by offering a standard definition of conceptual equivalence, and by suggesting that the adoption of a universalist orientation would require substantial changes to guidelines and more empirical work on the conceptualization of HRQOL in different cultures. PMID- 9226982 TI - Development of a tool to measure the life situation of parents of children with cancer. AB - The aim of this study was to test the validity and reliability of the recently developed Life Situation Scale for Parents (LSS-P) among parents of children with cancer. One hundred and ten parents of seventy-four children and adolescents who visited three paediatric wards in Sweden filled out three instruments: The LSS-P, the Quality of Life Scale and the Family Support Scale. The reliability coefficient, Cronbach's alpha, was found to be 0.82 for the LSS-P. A factor analysis with orthogonal varimax rotation of 37 items of the LSS-P gave twelve factors. A higher order factor analysis reduced the factors to four (Care, Well being, Social life and Preparedness), explaining the underlying dimensions to 57.9%. The total LSS-P correlated significantly with the Quality of Life Scale, and the higher order factor Care with the Family Support Scale. The LSS-P discriminated, in some aspects, between two-parent visiting the ward for treatment or check-up. The conclusion is that this first version of the LSS-P was valid and reliable (internal consistency) to a certain extent, but that the instrument should be tested on larger samples and during different phases of the disease. PMID- 9226983 TI - Management in general practice significantly reduced psychosocial consequences of female urinary incontinence. AB - Urinary incontinence is a common health problem among women, and a spectrum of psychosocial problems is associated with this disorder. We have investigated how psychosocial impact changed during a management programme for urinary incontinence in general practice. One hundred and five women seeking help for urinary incontinence were treated with conservative treatment options. Psychosocial consequences, grouped as mental distress (nine items), practical inconveniences (five items), and social restrictions (11 items) were noted before treatment, and after 3, 6 and 12 months follow-up. Urge symptoms, high degree of severity, and long duration were associated with higher psychosocial impact. During treatment, psychosocial impact was significantly reduced and the degree of impact in the three consequence groups was reduced to about one third compared with before treatment. In conclusion, changes in psychosocial impact during a management programme occur as a response to successful treatment. These findings support the view that female urinary incontinence can be successfully treated in general practice. PMID- 9226984 TI - General life satisfaction and domain-specific quality of life in chronic schizophrenic patients. AB - Subjective quality of life (QOL) has often been assessed through questionnaires or structured interviews focusing on the person's satisfaction with various life domains. In particular, most QOL instruments for psychiatric patients are based on this concept. We report on a study casting some doubts on the rationale of this approach. We investigated the QOL of 48 chronic schizophrenic outpatients with a long-term disease history (at least 20 years) using a German version of the Lancashire QOL Profile. The interrelations between general life satisfaction, satisfaction with specific life domains, psychological well-being and psychopathology were studied using correlation analysis and multiple linear regression. Of the life domains assessed, only two, namely social relations and health, contributed significantly to the patients' general life satisfaction, while the others (including work, leisure, family relations and housing) did not. The subscales on psychological well-being (self-esteem, affective state) as well as psychopathology were found to be more closely associated with general life satisfaction than almost all life domains considered. The findings are discussed with regard to the specific situation of the group of patients investigated. They give indications that the life domain approach to measuring QOL has its limitations, in particular when applied to patients having adapted to a very restricted everyday life. PMID- 9226985 TI - Performance and quality of life outcome in patients completing concomitant chemoradiotherapy protocols for head and neck cancer. AB - This study evaluated post-treatment performance and quality of life (QOL) outcome in head and neck cancer (HNC) patients treated with organ preservation, intensive chemoradiotherapy (FHX). Participants were 47 Stage II-IV HNC patients with no evidence of disease at least one year post-completion of organ preservation, concomitant FHX treatment. Patients were assessed via a semi-structured in-person interview, standardized measures of QOL (FACT-H&N, CES-D), performance (PSS-HN) and patients' perception of residual side effects. Disease, treatment and toxicity data were retrieved from medical charts and protocol records. The most salient performance impairment was inability to eat a normal solid food diet, with 50% of patients able to eat soft foods or take liquids only. This specific functional deficit was not related to global QOL, nor to specific quality of life dimensions. Dry mouth, the most frequent and severe residual effect, was not associated with outcome diet, depression or QOL. Residual pain, seen in only 15% of patients, appeared to influence both functional and QOL parameters as well as being a marker for other troublesome symptoms. Twenty-three per cent of patients were depressed; depression was associated with past problems related to alcohol abuse. Decreased QOL and increased depressive symptomatology were related to total number and severity of residual effects. The data highlight the importance of systematic study of QOL dimensions and caution against making assumptions about patients' experience of particular disease and treatment sequelae. PMID- 9226986 TI - Friend of the court. PMID- 9226987 TI - Who ya gonna call? PMID- 9226988 TI - A fine mess. PMID- 9226989 TI - Stop and count to 30. PMID- 9226990 TI - Monday's child. PMID- 9226991 TI - Intracytoplasmic sperm injection for treatment of the infertile male. AB - Intracytoplasmic sperm injection (ICSI) with in vitro fertilization represents one of the most significant advances in fertility technology. In this relatively new procedure, a single viable sperm is microinjected into an oocyte that has been extracted transvaginally. After fertilization occurs, the embryo is transferred into the uterus. This procedure now affords men who were previously thought to be irreversibly infertile the chance to initiate their own biologic pregnancy. However, because of the procedure's significant costs and its potential risk to the mother, careful selection of couples following a thorough male factor evaluation is mandatory. PMID- 9226992 TI - Linear basal cell carcinomas: report of multiple sequential tumors localized to a radiotherapy port and review of the literature. AB - Linear basal cell carcinoma, an uncommon variant of basal cell carcinoma, was first described in 1985. The cases of eight additional patients with this morphologically distinctive variant have been published subsequently. We describe a woman who sequentially developed three linear basal cell carcinomas located within her radiotherapy port. Including our patient, 12 linear basal cell carcinomas have been reported in 10 patients (4 men and 6 women) ranging from ages 40 to 77 years. The tumors were located most often on the head and neck (9 cancers); other sites involved the shoulders (2 cancers) and posterior thorax (1 cancer). All of the tumors were excised without recurrence. Various mechanisms have been postulated to account for this morphologic variant of basal cell carcinomas including a Koebner phenomenon or limited lateral spread of the cancer secondary to dermal fibrosis, or both. PMID- 9226993 TI - The role of excitotoxicity in organophosphorous nerve agents central poisoning. PMID- 9226994 TI - Schizophrenia and L-745,870, a novel dopamine D4 receptor antagonist. AB - The discovery of a novel high-affinity and selective dopamine D4 receptor antagonist, L-745,870, and the results of clinical trials with this compound are reviewed. Despite several lines of evidence which suggest that a selective D4 receptor antagonist may be an effective antipsychotic agent with a lower propensity to induce extrapyramidal side-effects, L-745,870 was ineffective as an antipsychotic in humans. PMID- 9226995 TI - Nitric oxide: inducer or suppressor of apoptosis? PMID- 9226996 TI - Fluoxetine hypophagia. Is there a role for serotonergic mechanisms in some circumstances? PMID- 9226997 TI - Pitfalls using metalloporphyrins in carbon monoxide research. AB - The proposal that endogenously produced carbon monoxide (CO) may act as a biological messenger has remained controversial. Carbon monoxide is generated by haem oxygenase isoenzymes in the degradation of haem-containing molecules. Certain metalloporphyrins, which are inhibitors of haem oxygenase, have been widely used as pharmacological tools in order to establish a messenger role for CO in the brain and periphery. However, increasing evidence shows that many metalloporphyrins are also associated with a large range of undesired effects, which make the interpretation of results using such compounds very uncertain. In this article, Lars Grundemar and Lars Ny evaluate the properties and describe the nonselective effect profile of such metalloporphyrins. PMID- 9226998 TI - Therapeutic advances in amyotrophic lateral sclerosis. AB - Amyotrophic lateral sclerosis (ALS) is a progressive and rapidly fatal neurodegenerative disease in which both upper and lower motoneurones are involved. The recent discovery of mutations affecting the superoxide dismutase (SOD) gene has given impetus to research on the role of oxidative stress in the pathogenesis of familial ALS, while further evidence for a role of excitotoxicity in the disease process has arisen. In this review, Erik Louvel, Jacques Hugon and Adam Doble discuss these findings and, in addition, describe how a number of large, well-controlled clinical trials have taken place to test potential therapies suggested by different aetiological hypotheses, including immunosuppressive therapies, neurotrophic factors, antioxidants and anti excitotoxic drugs. These trials have led to the first modest steps in the treatment of this devastating neurological disease. PMID- 9226999 TI - Selective inhibitors of neuronal nitric oxide synthase--is no NOS really good NOS for the nervous system? AB - It is now ten years since NO was shown to account for the biological activity of endothelium-derived relaxing factor (EDRF). It is also the tenth anniversary of the identification of L-NG monomethyl arginine (L-NMMA) as the very first inhibitor of NO biosynthesis. That EDRF and NO were one and the same sparked an explosion of interest in the biochemistry and pharmacology of NO which has yet to subside. In contrast, the first ever nitric oxide synthase (NOS) inhibitor slipped seamlessly into the literature virtually without comment at the time. Over the following decade, L-NMMA (and like NOS inhibitors) have proved invaluable as tools for probing the biological roles of NO in health and disease and, in particular, have increased our understanding of the function of NO in the nervous system. Further advances in this important area now require the development of inhibitors selective for the neuronal isoform of NOS (nNOS). Here, Philip Moore and Rachel Handy provide an up-to-date account of the literature regarding the biochemical and pharmacological characterization of NOS inhibitors with particular reference to compounds with greater selectivity for the nNOS isoform. PMID- 9227001 TI - Cephalopelvic disproportion. PMID- 9227000 TI - Gene targeting--homing in on alpha 2-adrenoceptor-subtype function. AB - The alpha-adrenoceptor was subdivided into three subtypes: alpha 2A-, alpha 2B- and alpha 2C-adrenoceptors almost ten years ago. Since then, the search has been on to discover and develop subtype-selective agonists and antagonists, but as yet no major breakthrough has been made. In the past year, several strains of genetically engineered mice have become available, either overexpressing, totally lacking or expressing heavily modified alpha 2-adrenoceptor subtypes. Ewen MacDonald, Brian Kobilka and Mika Scheinin describe how these mice may be utilized to elucidate the physiological functions of the receptor subtypes and the properties of future subtype-selective drugs. PMID- 9227002 TI - Histopathology services in a rural African hospital: how audit can improve service. AB - Audit is about doing things right. We undertook a detailed audit of the histopathological service at a rural district hospital in Africa because delays in obtaining biopsy results had been noted by clinicians. A wide range of serious pathology was found in the 100 consecutive biopsies reviewed. It took 26 days on average from the time a biopsy was taken to the time the result was returned to the clinician, and most of this delay was administrative in nature, occurring after the pathologist had reviewed the specimen at the regional laboratory. Because of these delays, only 22% of patients biopsied were ever informed of their results. By performing this audit, reporting the results, and acting against the problems discovered, the service rapidly improved. This study illustrates the importance of simple, routine audit in district hospitals in developing countries. PMID- 9227003 TI - Analgesia for severe postoperative pain: a comparison of two methods. PMID- 9227005 TI - Clinical presentation of pulmonary tuberculosis in under 10s and differences in AIDS-related cases: a cohort study of 115 patients. AB - One hundred and fifteen cases of pulmonary tuberculosis (PTB) in children under 10 were reviewed, including a case-control retrospective study between HIV positive (+ve) and HIV negative (-ve) children. Overall, respiratory symptoms not responding to acute respiratory infection (ARI) protocol and > 10% weight loss or failure to thrive during 3 months were the main presenting symptoms, but chronic fever alone is also common in HIV infected children with PTB. Hylar enlargement is the most frequent radiologic pattern, although lobar infiltrates are common when HIV infection coexists. Gastric lavage culture was an important diagnostic tool but Mantoux test, gastric lavage direct smear and erythrocyte sedimentation rate (ESR) levels, were not helpful in diagnosing PTB. Our findings suggest that when HIV infection is suspected or confirmed, chronic fever and lower lobe infiltrates should also be considered as PTB warning signs. PMID- 9227004 TI - Anthropometric measures as a predictor of cephalopelvic disproportion. PMID- 9227006 TI - Basic dermatological treatments for tropical district hospitals. PMID- 9227008 TI - Childhood chronic lung disease in the United Arab Emirates. AB - We report our observations on the pattern of referral of children with chronic lung disease (CLD) in Al Ain, United Arab Emirates. In a 1-year period 45 children were seen with severe lung disease from an estimated childhood population of 90,000. Bronchiectasis, cystic fibrosis (CF) and congenital lung disorders were the main diagnoses made. The indigenous Arab population who represent half the total population of the district appear to be at particular risk of severe lung disease. Chest X-ray and high resolution computerized tomography (CT) were the most commonly used imaging investigations to reach a diagnosis. PMID- 9227007 TI - Chloroquine-resistant Plasmodium falciparum among children in Calabar, south eastern Nigeria. AB - Sixty-nine children aged between 6 and 60 months with parasitologically proven Plasmodium falciparum malaria were treated with chloroquine (2.5 mg/kg) in the Children's Emergency Room of the University of Calabar Teaching Hospital (UCTH) in 1993. Thirty subjects (mean age 27.8 months) and 39 (mean age 29.5 months) received chloroquine phosphate suppository (Pharma Deko) and chloroquine sulphate syrup (May & Baker), respectively. The World Health Organization (WHO) 14-day in vivo field test was used in evaluating the response to treatment. In both treatment groups the responses were similar. Overall, parasitological cure occurred in 24 subjects (34.8%) and in the remaining 45 subjects (65.2%) treatment failed (chloroquine resistance). This level of chloroquine resistant Plasmodium falciparum (CRPF) is higher than 53.6% reported in this centre in 1989. Furthermore, in the present study the proportion of RII (46.4%) is significantly higher than 21.4% (P < 0.02) obtained in 1989. Our findings show a worsening of CRPF in Calabar with RII being the main contributor. This observation indicates the need for continued surveillance of the response of P. falciparum to chloroquine and alternative antimalarials as a means of evolving an effective treatment policy for malaria. PMID- 9227009 TI - General anaesthesia for cleft lip and palate surgery team activities in Cambodia. AB - Cleft lip and palate surgery team activities in Cambodia were launched in 1989 by a Japanese non-governmental organization. The objectives of the project were to provide appropriate surgical treatment and safe general anaesthesia for local patients, and also to teach general anaesthesia and surgery to local medical staff. A surgery team was sent on four occasions between 1991 and 1993 and a total of 130 patients received surgical treatment and general anaesthesia. Anaesthesia techniques employed included total intravenous anaesthesia in 70 patients and intravenous anaesthesia and low dose halothane in 60 patients. There were no major complications, such as airway obstruction or apnoea in the postoperative period. Total intravenous anaesthesia is an appropriate technique for patients in developing countries. The teaching of anaesthesia should be emphasized during the surgery team activities. PMID- 9227010 TI - Primary small bowel volvulus in rural Nepal. AB - Primary small bowel volvulus is the commonest cause of intestinal obstruction in the Gorkha district of Nepal resulting in laparotomy. Yet, this problem is mentioned only briefly, if at all, in many standard textbooks of surgery. This paper presents details of the presentation, clinical findings, and surgical management of 18 cases of primary small bowel volvulus. Small bowel volvulus occurred in adults only, with an overwhelming male predominance and a low mortality despite late presentation in the majority of cases. Attention is drawn to the common finding of localized ischaemia of several centimetres of terminal ileum. PMID- 9227011 TI - A community-based investigation of maternal mortality from obstetric haemorrhage in rural Zimbabwe. Maternal Mortality Study Group. AB - In the rural province Masvingo in Zimbabwe, 25% of maternal deaths were caused by obstetric haemorrhage, which had a cause specific maternal mortality rate (MMR) of 40 per 100,000 live births. Forty per cent of cases were due to a ruptured uterus, and 30% to an atonic uterus. Forty-two per cent were more than 35 years old and 44% para 5 or more. In spite of antenatal coverage for 85% of the women, 42% died outside any health facility. Fifty per cent of the women had had no intervention whatsoever before death from haemorrhage. The most important factor for prevention at community level is provision of emergency transport, which would have saved 50% of the women. Other non-health service factors contributing to the adverse outcome were found in actions of the patient herself or a traditional birth attendant. In the health services avoidable factors were identified in 58% of women. More effective antenatal attention to high risk factors, especially high age and parity, appropriate use of maternity waiting shelters, action programmes for management and haemorrhage at all levels, basic resources for resuscitation, improved surgical skills with supervision and available transport for referrals are all necessary parts of a programme to prevent maternal deaths from obstetric haemorrhage. PMID- 9227012 TI - How to bring surgery to remote tribal areas. AB - In a mission hospital located in a tribal area in India we found that surgical patients were not coming to the hospital due to a variety of reasons. Table 1 summarizes the main problems and how they were overcome. The effectiveness of our efforts was reflected in the increase in the number of operations after these measures were carried out (1022 before and 1865 after). PMID- 9227013 TI - Dermatology: a useful tool in a depressed economy. AB - Dermatology is an aspect of medicine which can help produce a cheap and readily available tool for diagnosis, follow-up and epidemiologic study in a depressed economy such as in the developing countries of Africa and Asia. This would involve the provision of some dermatological training to students of medicine, nursing and other paramedical specialities, to allow maximum utilization of dermatology at the primary, secondary and even tertiary health institutions. PMID- 9227014 TI - The vacuum extractor for newborn airway suctioning. PMID- 9227015 TI - Improvised cannulae for peripheral venous cutdown. PMID- 9227016 TI - Recurrent paratesticular rhabdomyosarcoma reported from Nigeria. PMID- 9227018 TI - Wound infection in a rural hospital: the benefit of a wound management protocol. PMID- 9227017 TI - Perinatal mortality in northern Benin. PMID- 9227019 TI - Jeep door handle injury. PMID- 9227020 TI - Routine prenatal screening of Indian women for HBsAg: benefits derived versus cost. PMID- 9227021 TI - Erectile failure and destruction of glans penis by tuberculosis. PMID- 9227022 TI - Inversion uterus with intestinal obstruction. PMID- 9227023 TI - Amoebic liver abscess communicating with intrahepatic bile ducts. PMID- 9227024 TI - Enterobius vermicularis (pin worm) causing symptoms of appendicitis. PMID- 9227025 TI - Complication of acute pancreatitis: management of a haemorrhagic pancreatic pseudocyst. PMID- 9227026 TI - An unusual presentation of tuberculosis. PMID- 9227028 TI - Presentation of parasites and the radiology of parasitic diseases. PMID- 9227027 TI - Ruptured uterus presenting as a rapidly growing cystic mass in the epigastrium. PMID- 9227029 TI - Calcific enterolithiasis. PMID- 9227030 TI - Percutaneous blood salvage prior to auto-transfusion. PMID- 9227031 TI - Tropical diabetes? PMID- 9227032 TI - Sensitivities of malaria in Zaire. PMID- 9227033 TI - The Humphrey ADE block. PMID- 9227034 TI - Cotton-wick method for better drainage of middle ear. PMID- 9227035 TI - Cephalic vein cutdown: a new approach. PMID- 9227036 TI - Researchers identify diabetes gene. PMID- 9227037 TI - Medical care for the underclass. PMID- 9227039 TI - Access to health care in Virginia. PMID- 9227038 TI - Community partnerships work. AB - This adult care program epitomizes the values most recently espoused in debates on health care reform. Public/private partnerships are strengthened by emphasizing unique contributions of all resources of health care: private physicians, private pharmacists, local government, local health departments, community hospitals, business leaders, volunteer free clinics and a private medical school. The success of this program is also contingent on a high degree of patient responsibility. Patients must demonstrate a willingness to share in their care through improved capabilities of self-management. They must share in the cost of their medications, and if there is a propensity to abuse the services (unnecessary ER usage, doctor hopping), they run the risk of termination of services. Expectations are high and patients are being taught how to meet them. The next step is an intensive evaluation of outcome measures. It is felt that this evaluation must extend over a minimum of 5 years in order to be valid and demonstrate trends. We are currently pursuing funding for such an evaluation. PMID- 9227040 TI - Improving access to care. Carilion Health System. PMID- 9227041 TI - Improving access to health care for all Virginians. Trigon Blue Cross Blue Shield. PMID- 9227042 TI - The Virginia Statewide Area Health Education Centers Program. PMID- 9227043 TI - The Virginia Health Care Foundation. Meeting the challenge. PMID- 9227044 TI - Community health centers. Providing access to care in Virginia's medically underserved areas. PMID- 9227045 TI - Two chairs and beyond: protecting our patients and our medical profession. PMID- 9227046 TI - The birth of emergency medicine. A retrospective. PMID- 9227047 TI - Esophagitis associated with alendronate sodium. PMID- 9227048 TI - Alcohol withdrawal. AB - There are three concurrent processes involved in the withdrawal from alcohol in an alcohol-dependent person. The first process is the hippocampal calcium channel mechanism diagnosed by the coarse tremor leading, sometimes precipitously, to convulsions. The second process is commonly referred to as alcoholic hallucinosis, and involves the psychoactive biogenic amine, harmine. The third process involves the locus coeruleus and presents as irritability, a fine tremor, autonomic storm, and diaphoresis. Magnesium and phenobarbital are usually sufficient to treat the syndrome of alcohol withdrawal, although neuroleptics may be required. PMID- 9227049 TI - Total knee arthroplasty in the octogenarian. The patients' perspective. PMID- 9227050 TI - The health practitioners' intervention program. PMID- 9227051 TI - Prepare to negotiate your next employment agreement. PMID- 9227052 TI - Anesthetizing the dental pulp. PMID- 9227053 TI - Pulp stimulation and pulp capping. PMID- 9227054 TI - Hidden root and root canal system curvatures: discernment and treatment strategies. PMID- 9227055 TI - The heat wave: the secret to creating obturation excellence. PMID- 9227056 TI - Common errors in periradicular surgery. PMID- 9227057 TI - Endodontic cleaning and shaping: current concepts. PMID- 9227058 TI - Morphometric study of teeth in age calculation. AB - The diagnostic usefulness of some morphometric parameters of teeth in age determination were studied. In the first series 173 central incisors from 13-83 year-old individuals were examined and in the second series 72 teeth from 30 individuals (aged 39 to 80 years). Root transparency and dentinal tubule diameter were the most reliable parameters on the basis of image analysis (IBAS system) of scanning electron microscopic images. These variables were used in multiple linear regression analyses which led to the exclusion of secondary dentine as it did not improve the fit and therefore did not help to explain the dependent variable. The precise measurement of the morphometric variables we used facilitates age determination, reducing the mean error reported by other authors who used subjective estimates. Our results document the limited effectiveness of these parameters in age estimation due to individual variations caused by genetic factors and chewing habits. PMID- 9227059 TI - Dental root surface structure as an indicator of age. AB - The purpose of this investigation was to examine the relationship between the root surface structure of human teeth and the age of the individual and also to evaluate whatever contribution this relationship might give to multiple regression methods for age estimation. The material, consisting of 1000 permanent teeth, excluding molars, was examined by means of a new scoring system (Surface Roughness Scores, SRS) for surface roughness, in addition to the scoring systems of Gustafson (RG) and Johanson (RJ) for root resorption. Statistical analyses using the SPSS package indicated a symmetric left/right distribution of root surface structure. The Pearson correlation between age and RJ varied from -0.02 for maxillary central incisors to 0.54 for mandibular central incisors and was approximately the same for RG. These two scoring systems seem to be of little value in methods for age estimation. Correlations between age and the new scoring systems SRS were significant for all teeth and varied from 0.44 for maxillary second premolars to 0.68 for mandibular first premolars. There was no detectable influence of gender or reason for extraction. However, the SRS could not be assessed with sufficient reproducibility, and the estimates were therefore too subjective to be used as the sole criterion for age estimation. This scoring system could, however contribute positively to a multiple regression method. PMID- 9227061 TI - Bitemarks in forensic dental practice: the Russian experience. PMID- 9227060 TI - Dental radiographic identification utilising computerised digital slice interposition: a case report. AB - A pair of ante- and post-mortem periapical dental radiographs of an individual were digitised on a personal computer with the aid of an X-ray scanner. The radiographs were appropriately prepared and compared using image editing software. In this case, dental restorations, root morphology, periodontal ligaments, pulp spaces, bony trabeculation and a retained root fragment were features used for comparison. The results showed that all the features were consistent in ante-mortem and post-mortem radiographs except for two points. Both discrepancies were related to dimensional differences found in dental restorations and the retained root fragment. Nevertheless, there were acceptable explanations to account for each discrepancy. PMID- 9227062 TI - Comparative review of bitemark cases from Pretoria, South Africa. AB - Bitemark evidence has become more scientifically based and is currently widely accepted in the legal process. Bitemarks can be inflicted by humans or animals on humans, animals and a variety of inanimate objects and can be found on any part of the body, with their quality and appearance being influenced by a variety of factors. The purpose of this study was to record the experiences with bitemark cases presented to forensic odontologists at the University of Pretoria from 1983 1993 and to compare them with trends and findings elsewhere. Sixteen cases are presented, of which 14 were bitemarks inflicted by humans and two by dogs. Thirteen cases occurred in human tissues, three in inanimate objects. Of the bitemarks in human skin, most were present on the arms, followed by the face, thorax and back. Bitemarks over the entire body were seen in the two victims bitten by dogs. The male:female ratio was 4:1 and in 46% of cases single marks were present while the rest were multiple. Eight of the victims had been assaulted. Two cases were associated with sexual behaviour (rape), two were inflicted by dogs, and the circumstances surrounding one case were unknown. A variety of factors complicated the investigations. The major factors responsible for disqualifying bitemarks as evidence included mutilation, removal of tissues, inexperience of officials involved and multiple bitemarks. The results of the study confirm the importance of bitemarks as forensic evidence. Humans are the primary victims with the arm being the anatomical site most often involved. Inexperience on the part of the investigating officers and other officials in the handling of these cases strongly emphasises the need for proper training and education of these personnel. PMID- 9227063 TI - Image editing and computer assisted bitemark analysis: a case report. AB - Bitemark evidence in a homicide usually involves a perpetrator biting the victim prior to or around the time of death. This paper presents a case in which a homicide victim bit his assailant. A suspect taken into custody was found to have what appeared to be a human bitemark on the proximal phalanx of his right thumb. Scale photographs of this injury were obtained and compared to the dentition of the decreased using digitized computer images for superimposition. Three different approaches for comparison with the bitemark photograph were utilized: comparison with radiographs of amalgam-filled impressions of dental casts, a transparent overlay technique and comparison with photographs of a simulated bitemark inked onto the hand of a volunteer. A review of these techniques as they apply to computerized bitemark analysis is presented. PMID- 9227064 TI - A bitemark case with a twist. AB - This is a case report in which the bite patterns of two suspects were compared to a bitemark on the breast of a murder victim. Each suspect had sufficient concordant features to have been found guilty of producing the bitemark. The irony in this case is that the bitemark was not inflicted by the murderer. PMID- 9227066 TI - Individualisation of dental tissue--an aid for odontological identification? AB - The introduction of new methods in forensic diagnostics, especially serological techniques, including the use of individual markers for identification is becoming increasingly important. The DNA techniques are particularly promising and dental tissue, especially dental pulp, is a good source of DNA because it is well protected against autolysis. Gc-subtyping and application of DNA techniques for identification were reported in 1992 and show the efficacy of PCR systems for the individualisation of dental tissues. In cases of optimal conditions-room temperature and dry air-the analysis was successful after 6 or 12 months and the results could be used for identification. Under the influence of high temperature autolysis occurs and the pulp degrades making DNA typing almost impossible. The experiments show that the system HLA-DQ alpha is more reliable than the system MCT 118 and the results confirm that these techniques can be used for identification of unknown persons in some cases. The methods are only usable if comparative material belonging to the subject, such as hair, is available. PMID- 9227065 TI - Comparison of bitemarks in foodstuffs by computer imaging: a case report. AB - Police called to investigate a fire in a snackbar in Mount Gambier, South Australia, discovered four cakes with characteristics marks apparently produced by human teeth. These marks were examined and compared with the teeth of a suspect arsonist. The comparison was made by computer imaging analysis and a remarkable similarity in arch shape was observed. PMID- 9227067 TI - Accuracy in identification of implant treated patients by use of intraoral radiographs. AB - This study aimed at investigating the accuracy of seven forensic odontologists (FO) and seven police officers (PO) in determining person identity using comparable intraoral radiographs from implant treated patients, and evaluating the different characteristics used for identification. The investigation was based on 34 edentulous patients, from 26 of whom a matched pair of radiographs was constructed in such a way that one, taken after insertion of an implant supported prosthesis in the anterior part of the mandible, was regarded as the antemortem radiograph, while another picture from a later follow-up examination served as the postmortem x-ray. From each of the remaining eight patients similar radiographs were selected so that four antemortem and four postmortem ones were obtained. These eight radiographs were also paired but did not match. There were thus 30 pairs of radiographs. A classification of the 26 matched pairs regarding degree of ease (easy, moderate, difficult) in combining the radiographs, using as parameters the design of the implants, shape of the abutments, shape of the bridges and bony anatomy of the jaws was established. The total number of errors made by the FO were higher (26) than those made by the PO (18) (one PO combined all 26 matching radiographs correctly) and 12 of the 26 matchable cases were correctly paired by all observers. The design of the fixed prostheses was the most often used characteristic in the exercise. PMID- 9227068 TI - In search of a suitable denture marker. AB - The ID-Band (SDI AB, Sweden) has become the standard, internationally and FDI accepted denture marking system. In Australia however the strip is not easily obtainable and is expensive. Two other materials have been trialled as possible alternatives: (1) Titanium foil (9 microns) and (2) Ho Band (matrix) (3 microns) (Lorvic Corp, USA). All three bands were tested for tensile strength and elongation at temperatures: RT, 700 degrees and 900 degrees C. As the ID- and Ho Bands are both 18-8 stainless steel their performance was similar. The 18-8 was stronger, had a higher percentage break point and a higher elongation. The latter meant that it was softer and could be more easily inscribed and was therefore more suitable for denture marking. T1 is becoming increasingly used in dentistry but in spite of its abundance it is not likely to replace stainless steel for denture marking at present. On the other hand the Ho Band is cheaper, more readily available and it could replace ID-Band for use in Australia. An alternative paper based marking system is also presented together with the rationale for its use. PMID- 9227069 TI - Denture marking. Clinical and technical aspects. AB - The frequency of edentulousness has decreased in recent years due to the improvement in oral health. However, there is still a need to address the issue of denture marking for social and legal reasons because the oral status of populations varies in different countries and the wearing of complete dentures will be a fact for the foreseeable future. Given that only one marked denture can tell us the identity of a decreased when all other methods fail makes it a worthwhile exercise. The marking of dentures is not regulated by law in Sweden, but it is recommended by the Swedish Board of Health and Welfare (SOSFS[M]1986), that all patients should be offered the opportunity to have their dentures marked, which they may refuse. In Sweden, the dental laboratories report that they mark all dentures. The Swedish ID-Band has become the international standard and FDI accepted denture marking system, but recent research has indicated that this metal band is not resistant to very high temperatures. Since there is no international consensus regarding the matter we suggest that new materials should be explored. PMID- 9227070 TI - Quantitative forensic evaluation of bite marks with the aid of a shape analysis computer program: Part 1; The development of "SCIP" and the similarity index. AB - Bite marks left on human tissue and bitten material have become an important aspect of scientific evidence used for the conviction or acquittal of a suspect. Expert opinion has often been based on subjective comparisons rather than any objective metrical analysis and many experts will agree that there is a need to employ additional comparative tests to achieve unbiased objectivity in their investigation. In this study, an interactive shape analysis computer program ("SCIP"-Shape Comparison Interactive Program) has been employed in an attempt to derive experimentally a quantitative comparison, in the form of a Similarity Index (S.I.), between the "offender's" teeth and the bite marks produced on a standard flat wax form. The S.I. values obtained using "SCIP" were evaluated in a variety of experimental bite mark situations. It was found that in no case could the S.I. values produced by comparison of the bite mark with the dental casts from non-perpetrators be confused with the much lower S.I. from comparison of the bite mark with the dental cast of the perpetrator. The use of the Similarity Index derived using the "SCIP" program is recommended as a simple, accurate and objective means of comparing bite marks in suitable forensic cases. PMID- 9227071 TI - Quantitative forensic evaluation of bite marks with the aid of a shape analysis computer program: Part 2; "SCIP" and bite marks in skin and foodstuffs. AB - In a previous paper 1, we have shown that the use of an interactive shape analysis computer program ("SCIP") and the derivation of a quantitative Similarity Index 1 greatly facilitated the comparison of experimental flat wax bite marks with the dentition of various 'suspects' and the identification of the agent producing the bite. In this study, "SCIP" was employed in an attempt to quantify the comparison, in the form of the Similarity Index (S.I.), between the "offender's" teeth and the bite marks produced on foodstuffs and on human skin, under experimental conditions. The use of "SCIP" and the S.I. is recommended as a routine means of eliminating suspects in bite mark cases. If a reasonable number of reference points have been registered in the bitten material and particularly if the perpetrator has any unusual features in the anterior dentition, the matching of the bite mark with the actual offender is a possibility with this method. PMID- 9227072 TI - A bitemark and a fracture? AB - The kidnap and brutal murder of the eleven year-old daughter of a fire brigade officer in the town of Wassenaar in the Netherlands on September 29, 1980, resulted in the first ever appearance of a forensic odontologist as an expert witness in the history of Dutch law. This previously unpublished case is now reviewed for its historic significance, and also because it presents an interesting problem of interpretation of odontological evidence relevant to the identification of the offender, and raises issues concerning proper procedures for the utilisation of expertise in forensic odontology. PMID- 9227073 TI - Scanning electron microscopy, a useful tool in forensic dental work. AB - In selected cases there is a need for microscopic information found on the surface of tooth specimens. Scanning electron microscopy is the method of choice. Based on the knowledge of the structural organization of the mineralized dental tissues differential diagnoses of physical and chemical changes can be made. It is suggested that the forensic dental profession cooperates in establishing a collection of reference material illustrating different traumas to enamel and dentin, deposits on teeth and the structure of restorative dental materials as they will appear after some years in position on a tooth. Four different cases are presented. PMID- 9227074 TI - Forensic implications of the variation in morphology of marginal serrations on the teeth of the great white shark. AB - The teeth of the Great White Shark have been examined to ascertain whether there is any commonality in the arrangement or number of the marginal serrations (peaks) or, indeed, whether individual sharks have a unique pattern of shapes or size of the peaks. The teeth of the White Shark are characteristic in size and shape with serrations along almost the entire mesial and distal margins. This study has revealed no consistent pattern of size or arrangement of the marginal serrations that was sufficiently characteristic within an individual shark to serve as a reliable index of identification of a tooth as originating from that particular shark. Nonetheless, the serrations are sufficiently distinctive to enable the potential identification of an individual tooth as having been the cause of a particular bitemark. PMID- 9227075 TI - Application of dimorphism in teeth to age calculation. AB - The objective of this study was to determine which teeth provide the most reliable data for use in age estimation. We studied 170 teeth from the anterior region (central and lateral incisors, and canines) and used an image analysis technique to analyse a number of morphometric features in thin sections of tooth. Our results showed root translucency to be the variable most clearly related to the subject's age, and the canine teeth to provide the most reliable information on this variable. PMID- 9227076 TI - The forensic implications of palaeopathology of the cranium and jaws. Part I. A technique for radiography of mummified and skeletal remains. AB - Radiography of dried skulls presents two major problems: the lack of soft tissue which usually leads to overexposure, and difficulty in maintaining the cranium, with or without mandible, in correct position. The first problem can be alleviated by suspending a one litre drip bag containing Hartman's solution across the X-ray tube head and angled to give a fluid thickness of 10 cm. Satisfactory positioning of the skulls and relating the mandible to cranium was achieved by construction of a spinal column substitute and acrylic hooks and rubber bands. Exposure of 50 KV and 10 mA at 15 seconds for orthopantomographic views and 70 KV at 1.5 seconds for lateral and 65 KV at 1.5 seconds for postero anterior views gave an optimal, clinically acceptable image. The perfused fluids of living tissues exert a strong, modifying influence on the X-ray beam. PMID- 9227077 TI - Teeth as a poison depot. 1954. PMID- 9227078 TI - Comparative study of experimentally induced and post-mortem pink teeth. AB - Forty-eight pink teeth from eight male cadavers, all dying from unnatural causes were studied for the pink tooth phenomenon. Perfusion with whole and lysed blood, at different temperatures was carried out, followed by longitudinal sectioning for histological observation. Another 30 unblemished human teeth were used to reproduce the phenomenon in vitro to help clarify its pathogenesis. PMID- 9227079 TI - Non-acceptance of prosthetic appliances at the focus of forensic consequences. AB - Non-acceptance of prosthetic appliances is not only a clinical problem; it also gives rise to legal proceedings. Not every case of non-acceptance is psychogenically induced, there are often somatic and even iatrogenic causes. Objective treatment deficiencies are subject to forensic sanctions irrespective of any underlying psychosomatic illness. If the dentist is to manage non acceptance in a responsible way not open to forensic consequences, competent clinical and psychological skills are needed. PMID- 9227080 TI - Third molars in the establishment of adult status--a case report. AB - Teeth are the most durable structures in the human body. The timing and sequence of their development, as contained in dental development charts, have been used as valid criteria for age determination. The third molars however are the last teeth to erupt and are regarded as the most variable in the dentition. Age estimation in a legal context, using developing third molars must be carefully applied otherwise justice may miscarry. A case of wrongful use of the technique is presented here. PMID- 9227081 TI - Identification of a suicide victim by facial reconstruction. AB - Facial reconstruction is used in an attempt to identify an individual by a three dimensional representation of the facial features using the skull as the foundation after metrical and non-metrical analysis to determine age, race and gender. The skeletonized remains of a female who was reported missing six years previously were recovered from the summit of Table Mountain in Cape Town. Some personal possessions were also recovered, one of which was a shark tooth pendant which the victim's parents recognized. Although there were distinctive dental features, the antemortem dental records had been lost during the initial investigation which therefore precluded identification by this means. However, positive identification was required and facial reconstruction on the skull was undertaken which the parents duly identified. The method used for facial reconstruction is described. PMID- 9227082 TI - A quantitative analysis of subtraction images based on bite-wing radiographs for simulated victim identification in forensic dentistry. AB - The comparison between ante- and post-mortem radiographs constitutes an important basis for victim identification in forensic dentistry. Due to the decline in dental caries among children that has occurred in Western countries over the last 20 years, the number of restorations in future populations will be limited. It is likely that this will impede successful victim identification in the future. It was the aim of this study to evaluate a new radiographic technique, subtraction radiography based on bitewing radiographs and determine whether it could provide an objective quantitative measure for victim identification. Bite-wings randomly sampled from a large population of adolescents (13-19 yrs) were video-camera recorded and digitized. Subtraction images were performed of pairs of bite-wings originating from the same individual (identical images) and from different individuals (non-identical images). The images were separated into two groups, one without fillings (12 identical and 48 non-identical) and one with simple amalgam fillings (15 identical and 60 non-identical). The distribution of grey shades in the subtraction image was used as the parameter for evaluation of homogeneity in the images. Subtraction images derived from identical radiographs were significantly more homogeneous than those derived from non-identical radiographs (p < 0.001) in both group with and without fillings. Although the difference was statistically highly significant, the distribution of grey shades in the two groups overlapped. Thus, the grey shades in the subtraction image cannot per se unequivocally establish the identity of a victim, but may add to the subjective comparison of two radiographs in victim identification. PMID- 9227083 TI - A non-destructive dental method for age estimation. AB - Dental radiographs have rarely been used in dental age estimation methods for adults and the aim of this investigation was to derive formulae for age calculation based on measurements of teeth and their radiographs. Age-related changes were studied in 452 extracted, unsectioned incisors, canines and premolars. The length of the apical translucent zone and extent of the periodontal retraction were measured on the teeth while the pulp length and width as well as root length and width were measured on the radiographs and the ratios between the root and pulp measurements calculated. For all types of teeth significant, negative Pearson's correlation coefficients were found between age and the ratios between the pulp and the root width. In this study also, the correlation between age and the length of the apical translucent zone was weaker than expected. The periodontal retraction was significantly correlated with age in maxillary premolars alone. Multiple regression analyses showed inclusion of the ratio between the measurements of the pulp and the root on the radiographs for all teeth; the length of the apical translucency in five types; and periodontal retraction in only three types of teeth. The correlation coefficients ranged from r = 0.48 to r = 0.90 between the chronological and the calculated age using the formulae from this multiple regression study. The strongest coefficients were for premolars. These formulae may be recommended for use in odontological age estimations in forensic and archaeological cases where teeth are loose or can be extracted and where it is important that the teeth are not sectioned. PMID- 9227085 TI - The Helderberg air disaster--forensic odontological investigations. AB - A Boeing 747-224B Combi of the South African Airways, the "Helderberg", crashed into the sea near Mauritius on 28 November 1987. All 159 people on board died and dental tissues were present in only eight of the 15 lots of human remains recovered. Ante-mortem dental records were collected by a team in Johannesburg while the post-mortem examinations were conducted in Mauritius. The special circumstances surrounding an accident at sea resulted in the low number of bodies available for identification procedures. Of the eight remains which included dental tissues, five were identified by means of simple dental restorations, advanced dentistry, anatomical features of teeth and stages of development of teeth. One of the victims was identified by a process of exclusion and radiographic evidence played a decisive role in the identification process. (J Forensic Odontostomatol 1994; 12: 15-18) The variety of record-keeping styles and abbreviations used in different countries posed a major problem during the process and it is concluded that international standardization in record-keeping requires urgent attention. PMID- 9227084 TI - Accuracy of dental registrations in forensic odontology among dentists and dental students. AB - In forensic odontology, registration of dental characteristics is crucial in the identification procedure. It has been found that the most common errors made are incorrect registration of restorations and confusion about premolars and molars in both jaws. In an earlier study, dental students were observers and the charting was made without radiographs. However, in practical forensic work dentists make the registrations and radiographs are usually available. In this investigation eight dental students and eight dentists made registrations on ten excised macerated jaws with the aid of radiographs. The mean number of errors for each jaw for the students and the dentist was 4 and 3 respectively. The most common error among the dentists was incorrect registration of restorations, while errors on registrations of missing teeth were most common among the students. Even though the material in this study was limited, the results indicate the importance of re-examining of postmortem findings before the comparison with the antemortem data is done. Additionally, the forensic work should be performed by specialists. PMID- 9227086 TI - The duty of the dentist to keep records--significance and relevance according to German law. AB - A dentist is duty-bound to keep dental records. The keeping of accurate records is in fact a legal obligation and an integral part of the medical or dental contract which can be used as evidence in court and a patient has the right to inspect and to take possession of the objective information contained in the records. Incomplete or inadequate records lead to a reversal of the burden of proof in German jurisdiction. PMID- 9227087 TI - The clinical management of gingival recession. PMID- 9227088 TI - An introduction to the psychological concept of control and its application to the dental setting. PMID- 9227089 TI - Does fluid intake affect the health of the periodontal tissues? An interesting case report. PMID- 9227090 TI - Periodontal assistance for developing nations. PMID- 9227091 TI - Periodontal conditions in HIV/AIDS patients. A short review. PMID- 9227092 TI - Smoking and periodontal disease. A review. AB - From the many epidemiological and clinical studies we have looked at there is considerable evidence supporting a positive association between smoking and periodontal disease. In conclusion therefore we can see that smoking can cause a number of changes within the periodontium which can predispose an individual to the progression of periodontal disease. Both the frequency and severity of periodontal disease appears to be greater among smokers than non-smokers. The smoking habit is associated with a variety of deleterious changes in the oral cavity. The periodontal tissues are firstly compromised by the initial vasodilatation and then the decreased blood flow to the gingiva, due to the vasoconstricting actions of nicotine. This also acts to decrease the crevicular fluid flow, which increases the susceptibility of the host to bacterial growth. The vasoconstriction also acts to inhibit what are normally early signs of periodontal problems by decreasing gingival inflammation, redness and bleeding. Evidence suggests that smoking is consistently associated with poorer levels of oral hygiene and consequently smokers have increased accumulations of plaque and calculus. The immune response of smokers is also impaired where the chemotactic and phagocytic ability of combination of these factors which enhances the breakdown of periodontal tissues in smokers. I believe that the dentist has an important role in making patients aware of the detrimental effects that smoking has on periodontal tissues and health in general. The smoking habits of all patients should be inquired about during an oral examination and the patient should be strongly advised to stop smoking by pointing out the risks involved and the positive benefits gained from breaking this habit. It is essential that the dentist is supportive in this role in an attempt to improve the oral status of the individual. PMID- 9227093 TI - Oral blood blisters: angina bullosa haemorrhagica (ABH). PMID- 9227095 TI - The past, present and future for dental hygienists. PMID- 9227094 TI - Peripheral odontogenic fibroma with clear cell odontogenic epithelium. AB - The clinical and histological features of the peripheral odontogenic fibroma are briefly outlined. A case arising from the attached lingual gingiva between the mandibular right permanent first molar and the second molar in a 67 year old Indian female is reported here. The unusual occurrence of marked clear cell differentiation within the odontogenic epithelial component, and histogenetic link to the clear cell rests of the dental lamina and surface epithelium are discussed. PMID- 9227096 TI - Reviewer selection: emphasis on expertise. PMID- 9227097 TI - Use of tissue-tinted porcelain to restore soft-tissue defects. AB - Anterior, partially edentulous residual ridges often have hard-tissue and soft tissue defects that create esthetic and functional problems when conventional fixed prostheses are used to replace the missing teeth. However, tissue-tinted porcelain can be used in conjunction with fixed prostheses to solve many of these problems. Proper prosthesis design and shade matching are important considerations in the fabrication of fixed prostheses that incorporate tissue tinted porcelain to restore hard-tissue and soft-tissue defects. PMID- 9227098 TI - Management of the maxilla after alveolar ridge augmentation with hydroxylapatite when opposed by mandibular implants. AB - Maxillary ridge augmentation and vestibuloplasty are often adjuncts to mandibular implant reconstructions. Careful management of the augmented ridge after surgery is critical to the success of the maxillary prosthesis and osseointegration of the mandibular implant. A technique is described for alteration of an existing maxillary complete denture after ridge augmentation. Precautions regarding prosthesis function during the osseointegration phase of different types of mandibular reconstructions are noted. Technique and rationale for denture flange modification when a prosthesis is rebased for a patient who received simultaneous vestibuloplasty and ridge augmentation are described. PMID- 9227099 TI - Modified bar superstructure for an implant-retained orbital prosthesis. AB - The success of a bar superstructure for an orbital prosthesis may be compromised by the placement and angulation of implants. The following technique describes a modification to a bar superstructure that provided the advantages of convenience, security, and consistent positioning even though one implant was lost and the angulation of implants limited accuracy. PMID- 9227100 TI - Common errors in panoramic radiography of edentulous patients. AB - PURPOSE: Instructions for patient positioning during panoramic radiography usually describe positioning dentate patients, and errors in patient positioning are commonly identified by distortion of the dentition. The purposes of this study were to identify common errors in panoramic radiography of edentulous patients, describe the image distortions that can be expected with positioning errors in edentulous patients, and review quality assurance methods that improve the diagnostic value of panoramic films. MATERIALS AND METHODS: Panoramic films were randomly selected from the inactive files of 75 edentulous patients seen at the dental school for complete denture construction. The radiographs were numbered and reviewed by a board-certified oral and maxillofacial radiologist trained to identify errors in panoramic radiography. RESULTS: Of the 75 panoramic radiographs examined, only 6 films (8.0%) were free of errors, and 67 films (89.3%) had one or more errors in patient positioning and 33 films (44.0%) had one or more technical errors. The most common positioning errors were positioning the chin too high (32 films, 41.3%) and positioning the patient too far forward (26 films, 34.7%). CONCLUSIONS: Without modification, manufacturer's instructions for positioning dentate patients during panoramic radiography may result in positioning errors on the panoramic radiographs of edentulous patients. Proper training and appropriate attention to detail while exposing and developing panoramic films are required to ensure maximum diagnostic benefits for edentulous patients. PMID- 9227101 TI - The relationship between preparation convergence and retention of extracoronal retainers. AB - PURPOSE: Although traditional ideal convergence (the sum of taper of the opposite sides) for crown preparation has been arbitrarily set at 4 degrees to 10 degrees, some believe absolute parallelism yields the highest retention. This study examined the relationship between the degree of convergence of a machined metal die and the retention of its casting. MATERIALS AND METHODS: The method used was that of cementing cast metal crowns onto full crown preparations on brass dies with varying convergence angles, and then recording the force required to remove the crowns from the dies in a vertical direction using a Tate-Emery Testing Machine and Load Indicator. RESULTS: It was found that retention (i.e., the force needed to remove the cemented castings from the die in their common long axis) increases from 0 degree convergence to peak between 6 degrees to 12 degrees convergence. It also seems that a critical film thickness does exist for optimum retention, and that film thicknesses smaller than the critical thickness may be responsible for the phenomenon that we have observed and directly related to the convergence angle itself. CONCLUSIONS: There seems to be experimental data supporting the use of traditionally taught convergence. Our study found that convergence angles between 6 degrees and 12 degrees seem to be optimum for tooth crown preparation when one plans to use zinc phosphate cement. Convergence angles of less than 6 degrees may not be desirable even if they can be clinically achieved. The results of our study indicate that a relationship exists between the convergence angle and the critical cement thickness that is necessary to realize the maximum strength properties of zinc phosphate cement. PMID- 9227102 TI - Adhesion promotion between metadent and a high palladium alloy with a pyrolytically fused porcelain opaque layer. AB - PURPOSE: Several authors have reported a preference for the use of high palladium alloys in the construction of implant-fixed partial denture (IFPD) substructures. Heat-polymerized polymethyl methacrylate resins (PMMR) are used to secure denture teeth to IFPD substructures. Metadent (Sun Medical Co, Kyoto, Japan), a heat polymerized PMMR containing 5% 4-methacryloxyethyl trimellitate anhydride (4 META), could possibly improve the overall design of the prosthesis because this resin adheres to resin and ceramic denture teeth and noble metals. However, adhesive bonding between Metadent and elemental palladium has been shown to be poor. The purpose of this study was to evaluate the potential for adhesive bonding between Metadent, titanate primers, and a ceramic layer pyrolytically fused to a high palladium alloy. MATERIALS AND METHODS: Five groups of 10 specimens were prepared. A layer of opaque porcelain was baked on all of the specimens under vacuum at 1760 degrees F. The specimens were grouped as follows: (1) Metadent resin processed on ceramic with no primer, (2) Metadent resin with 10% ethyl acetoacetate titanate (DC) processed on ceramic with no primer, (3) Metadent resin with 10% DC processed to ceramic with a 2% tetraisopropyl (TPT) surface primer, (4) 4-Metadent resin with 10% DC processed to ceramic with a 2% isopropyl dimethacryloyl isostearoyl titanate (KR-7) surface primer, and (5) Metadent resin with 10% DC processed to ceramic with a titanium dioxide layer on the opaque and a KR-7 surface primer. The primers were in a methyl methacrylate (MMA) solution. All of the specimens were heat-polymerized under pressure at 212 degrees F for 1 hour. One-half (25) of the specimens (5 from each group) were hydro-thermocycled at 4 degrees C to 55 degrees C, 0.5 minute dwell per bath for 3,000 cycles. All specimens were tested in shear on an MTS Universal machine at cross head speeds of 0.5/mm/min. RESULTS: The results showed adhesion between the resin-ceramic layer in virtually all groups regardless of thermocycling effects. The control (heat cure without 10% DC and no primer) and the 10% DC with KR-7 and TPT primer groups had an average MPa of 7.5, 7.7, and 7.0, respectively. The failure mode of the specimens was adhesive, with the site of failure being the porcelain-metal interface. CONCLUSION: It was concluded that adhesion between palladium alloy and Metadent was possible when a ceramic layer was used as an intermediary interface. The limitation seemed to be the adhesion of the ceramic to the metal. PMID- 9227103 TI - Passive fit of implant-retained prosthetic superstructures improved by electric discharge machining. AB - PURPOSE: An absolutely passive fit at the interface with the superstructure and the abutment cylinders is needed for implant longevity. In this study, a method of cast framework correction using electric discharge machining (EDM) was evaluated. MATERIALS AND METHODS: An original research model incorporating strain gages was devised to measure the fit of implant superstructures in three dimensions as cast and to compare this relationship with the fit following framework machining with EDM. RESULTS: Visually, the fit of the frameworks was excellent, and the results of ANOVA demonstrated that the mean fit of the frameworks significantly improved (P > .04) after machining with EDM. CONCLUSIONS: The EDM process offers a rapid, accurate, and simple technique to correct for casting inaccuracies in the fabrication of hybrid implant prostheses. PMID- 9227104 TI - The use of a photoprotective agent to increase the color stability of a tinted extraoral prosthetic silicone. AB - PURPOSE: This study investigated the use of ultraviolet-absorbing or photoprotective agents as a method of decreasing the color changes caused by ultraviolet radiation on an intrinsically tinted facial prosthetic material. MATERIALS AND METHODS: Silastic MDX 4-4210 (Dow Corning Corp, Midland, MI) was the facial prosthetic material studied. It was intrinsically tinted with talc and nylon flock to approximate the color of skin. Three brands of commercially available sunscreens with sun protective factor (SPF) of 15 were placed on the surface of the cured silicone samples. The samples were rotated in an ultraviolet radiation chamber for 300 hours. A control group was placed in the dark for 300 hours. The second phase of the experiment had para-aminobenzoic acid (PABA) added to the tinted silicone. All samples were measured with a spectrophotometer at the beginning and at the end of 300 hours, and a delta E was obtained. RESULTS: None of the sunscreens provided any ultraviolet radiation protection to the silicone. The addition of PABA caused a significant color degradation of the silicone. CONCLUSIONS: No photoprotective agent tested provided any significant level of ultraviolet radiation protection for silastic MDX 4-4210. PMID- 9227105 TI - In vitro analysis of the effects of two air-abrasive prophylaxis systems and inlet air pressure on the surface of titanium abutment cylinders. AB - PURPOSE: The purpose of this study was to investigate the effects of two air abrasive prophylaxis systems and the effect of inlet air pressure on the surface of Branemark titanium abutment cylinders. MATERIALS AND METHODS: Single abutment cylinders were treated with either the Prophy-Jet system (sodium bicarbonate abrasive) (Dentsply International, York, PA) or the Microprophy system (aluminum oxide abrasive) (Danville Engineering Co, Danville, CA) for 60 seconds at an inlet air pressure of 60 psi or 90 psi. The effects on the surface of each abutment cylinder were visually inspected by scanning electron microscopy. RESULTS: A comparison of abutment cylinder surfaces after treatment showed that the Prophy-Jet system removed machining marks to a greater degree than the Microprophy system. Sodium bicarbonate particles from the Prophy-Jet system were significantly larger than the aluminum oxide particles used with the Microprophy system, potentially accounting for the difference in abrasivity. In addition, inlet air pressure of 60 psi caused removal of machining marks to a greater degree than an inlet air pressure of 90 psi. The principle of phase separation may account for the lower inlet air pressure causing more removal of machining marks than the higher inlet air pressure. CONCLUSIONS: Under the experimental conditions tested, neither of the two systems tested seemed to cause significant abrasion of the surface of titanium abutment cylinders. PMID- 9227106 TI - A comparison of two silicoating techniques. AB - PURPOSE: A major challenge of composite-coated metal restorations is creating a strong bond between the two materials. This study was conducted to compare the bond strengths between composite and metal alloys using two silicoating treatments; the Silicoater Classic and the Silicoater MD (Kulzer Inc, Irvine, CA). MATERIALS AND METHODS: The Classic uses a pyrolytically applied silica glass and the MD process is an oven sintering of a metal oxide silicate layer. Two types of specimens of three different alloys were cast (Deva4: 51Au38Pd, G-Cast: 50Au32Ag12Cu [Degussa Corp. South Plainfield, NJ]; Ticonium: 70Ni15Cr [CMP Industries, Inc, Albany NY]). The specimens were silicoated with either the Classic or the MD process and opaque resin and composite (Dentacolor, Kulzer Inc) were bonded to each surface. A total of five control and 10 thermocycled (5,040 cycles 0 degree C to 67 degrees C; dwell time, 1 minute) specimens were fabricated for each alloy and specimen type. Two bond strength tests were used; a three-point flexure test and a shear bond strength test. Data were analyzed using a multivariate analysis of variance (MANOVA) and a Tukey's multiple comparison test. RESULTS: The standard deviations of the shear bond strength values ranged from +/- 2.30 to +/- 17.82.kg; thus, there was no significant difference in these results. Bond strength ratios were calculated from the flexure strength data. After thermocycling, the Silicoater MD produced significantly higher (P < 0.05) bond strength ratio values compared with the Silicoater Classic for Deva-4 (MD, 9.6 +/- 1.5 v Classic, 4.7 +/- 0.9) and G-Cast (MD, 11.6 +/- 1.2 v Classic, 3.9 +/- 0.5) but lower values for Ticonium (MD, 4.5 +/- 0.4 v Classic, 6.9 +/- 0.9). CONCLUSION: The Silicoater MD produced higher bond strength ratio values with noble alloys, and the Silicoater Classic produced higher bond strength ratio results when base metal alloys were used, suggesting a possible correlation between the effectiveness of the coating processes and the compositions of the alloys. These findings suggest that, for optimum results, selection of the type of silicoating process should be based on the components of the alloy. PMID- 9227107 TI - Cantilever and implant biomechanics: a review of the literature, Part 2. AB - In Part 2 of this literature review, a summary of the literature regarding the determination of acceptable cantilever lengths for fixed implant prostheses is presented. Studies examining the possible effects of biomechanical stress on both the implant prosthesis and the supporting bone are also discussed. PMID- 9227109 TI - An update on the non-surgical treatment of periodontal disease. AB - For the resolution of gingivitis, the mechanical therapy of scaling and polishing in combination with an efficient plaque control system remains the most efficient therapeutic package. Topical antimicrobial, particularly mouth rinses may aid in plaque control, and irrigation devices enhance their efficacy but are not mandatory to the treatment. In the case of periodontitis, mechanical therapy in addition to the use of antimicrobial agents have proven to be highly efficient, with successful clinical parameters achieved as endpoints. Because of the strong bacterial component of the disease process, its elimination with chemotherapeutic agents becomes a reality. Delivery systems of antimicrobials to the site of action have large potential as adjunct to the existing modalities of treatment. This will especially be applicable in moderate to severe cases where access is difficult. Future directions in research lead us to the potentials of non steroidal anti-inflammatory agents that interfere with prostaglandin metabolism. Possible alteration of the periodontal disease process at some point in time may be a reality. Inevitably, at some point, especially in severe cases, surgical intervention becomes a necessity as no mechanical therapy is possible because of inaccessibility. In which case, the point of contention is that for periodontal disease, sound principles of anti-infective therapy must be employed constantly. PMID- 9227108 TI - Histological and immunochemical studies of oral leukoplakia: phenotype and distribution of immunocompetent cells. AB - The Phenotype and distribution of immunocompetent cells in oral leukoplakia with different levels of dysplasia were analyzed. Cells were identified in two compartments of the oral mucosa, the epithelium and subepithelial connective tissue. One hundred cases of neutral-buffered formalin-fixed paraffin embedded biopsy materials including 10 cases of acetone-fixed frozen tissue sections were studied immunohistochemically. In the main lymphoid population of each groups, the T lymphocytes predominated over the B lymphocytes. The lymphoid cells were present either as diffuse aggregates or organized in follicular patterns with or without germinal center-like structures. When present, B lymphocytes were seen to constitute the above mentioned structures. T lymphocytes made up the paracortical areas. A decrease in CD4/CD8 ratio was observed in cases with severe dysplasia. Specimens classified as mild to severe dysplasia presented a significant increase in the number of CD1a (+) dendritic Langerhans cells when compared with those of epithelial hyperplasia. A significant increase in macrophage count was also obtained in the subephitelial connective tissue of all dysplastic cases. A significant increase of CD57 (+) natural killer/killer cells in the subephitelial connective tissue and HLA-DR expression by the keratinocytes was observed in cases with severe dysplasia. Correlation and analysis of the results revealed an immunocellular reaction that varied according to the degree of dysplasia in oral leukoplakia. Immunologic events, i.e. decreased CD4/CD8 ratio, increased density of natural killer/killer cells and HLA-DR expression by keratinocytes, occurring simultaneously in severe dysplasia are speculated to be indicative of early malignant transformation. PMID- 9227110 TI - The oral health of diabetics. PMID- 9227111 TI - The Miswak as an aid in oral hygiene. AB - The Miswak is chewing stick used throughout the Middle East. In this article for FDI World, Drs. John Hardie and Khaled Ahmed, both practitioners in Saudi Arabia, review the literature on its mechanical cleansing action and the chemicals within its fibres which may be more effective in promoting oral health. They speculate that, since the plaque removing properties of the miswak and conventional toothbrush are similar, the beneficial effects of the toothbrush may also depend less upon the mechanical efficiency of its bristles and more upon the chemical constituents of the toothpaste. PMID- 9227112 TI - Review of failed implant cases. AB - Five Failed implant cases were presented and gained the following; 1) The use of computed tomography is recommended for treatment planning such as to ascertain the three-dimensional position of the mandibular canal, maxillary sinus and nasal floor, and also to decide the diameter, length and inclination of the implant placement. 2) The implant should be located for enough from mental foramen, mandibular canal, maxillary sinus and nasal floor. 3) The use of surgical stent is recommended for accurate drilling of the implant site. 4) Adequate number of implants and strategic arrangement of implants are required to equally distribute stress. 5) An implant abutment which is connected to natural teeth will be affected by the difference of micromobility of the natural teeth and the trouble of abutment of prosthesis. PMID- 9227113 TI - A literature review on some effects of translation, rotation and dental eruption on mandibular growth relative to the maxilla. PMID- 9227114 TI - Bone growth factors: potential for use as an osseointegration enhancement technique (OET). PMID- 9227115 TI - Observations on the recurrence of destructive periodontal disease during a period of four years after treatment. AB - The upper and lower extreme percentiles of change in clinical variables were analyzed for a period of four years after completion of periodontal therapy in order to assess the patterns of disease recurrence after treatment. This analysis has demonstrated periods of healing and relapse of sites within individuals, among different individuals, and among different clinical assessment variables utilized. As not all of the sites within different individuals heal or relapse at the same rate, the random asynchronous burst model of pathogenesis seems appropriate to incorporate the different clinical assessment parameters and different sites in different individuals simultaneously. Long-term recession and pocketing, however, may demonstrate opposite trends, giving the erroneous interpretation that attachment levels remain constant with time, whereas, in fact, an active "dynamic equilibrium" of tissue remodeling is established. A statistical model based on the dynamic equilibrium concept would enable account to be taken of the multifactorial interactions that incorporate site and subject interrelationships between different clinical assessment parameters. The dynamic equilibrium model may be suitable therefore as the basis on which statistical analysis of the progression of periodontal disease may be considered. However, as the value of this or other models of the periodontal disease process have not been established, the analysis of data to derive meaningful conclusions from complex statistical techniques may be premature. PMID- 9227116 TI - Is the cart before the horse? PMID- 9227117 TI - Buonocore Memorial Lecture. The amalgam-tooth interface. PMID- 9227118 TI - State of the art of tooth-colored restoratives. PMID- 9227119 TI - Enhancement of resin bonding to heat-cured composite resin. AB - The purpose of this study was to evaluate the effectiveness of various surface treatments used to enhance the bond strength of resin cements to two different laboratory-processed composite resins. Seventy specimens of a microfilled composite resin (Concept) and 70 specimens of a micro-hybrid composite resin (Herculite XRV) were fabricated in metal wells and subjected to heat (250 degrees F) and pressure (85 psi) curing. An additional 70 specimens of each material were fabricated in the shape of disks and also subjected to the same heat/pressure curing. All composite resins were subjected to one of seven treatment regimens. The like-treated specimens were then bonded together using dual-curing resin cement and a uniform seating force (106 gm). After 7 days, bonded specimens were thermocycled 1000 times at 5 and 55 degrees C, and debond shear strengths were determined on a Universal Testing Machine. The use of microabrasion (50 microns aluminum oxide at 60 psi) and ceramic layer deposition (30 microns aluminum oxide with a ceramic additive at 75 psi) consistently improved the shear bond strength of the resin cements to both composite resins. The other treatment combinations provided varying effects. In conclusion, microabrasion or ceramic layer deposition are preferred methods to enhance the bond of resin cements to composite resins. PMID- 9227120 TI - Antibacterial activity of dentin bonding systems, resin-modified glass ionomers, and polyacid-modified composite resins. AB - The antibacterial effects of the dentin bonding systems Syntac, ProBOND, Gluma 3 Step, the resin-modified glass ionomers Photac-Fil, Fuji Lining LC, Fuji II LC, and the polyacid-modified composite resins VariGlass, Geristore, and Infinity were evaluated using the cariogenic bacteria S mutans, L salivarius, S sobrinus, and A viscosus in vitro with a modified cylinder drop plate agar diffusion assay. All glass ionomers, the polyacid-modified composites, and the primers and adhesives of the dentin bonding systems exhibited various degrees of antibacterial activity against most of the test bacteria. The antibacterial activity of the adhesives of dentin bonding systems was anticipated because of the glutaraldehyde used in their formulations. However, the antibacterial activity of the various primers was unexpected and indicates a dual antibacterial action of these systems. PMID- 9227121 TI - Dentistry's friend: calcium hydroxide. PMID- 9227122 TI - In vitro caries inhibition effects by conventional and resin-modified glass ionomer restorations. AB - The ability of a material to inhibit recurrent caries formation is an important clinical therapeutic property. The objectives of this study were to develop an initial anticariogenicity profile (from fluoride release, to fluoride uptake, to resistance to an artificial caries challenge), testing the ability of conventional versus resin-modified glass-ionomer restorations to resist decay, and to study the effect of using intermediary dentin bonding agent components on the development of surface and wall carious lesions adjacent to a resin-modified glass-ionomer restoration. Cumulative fluoride release was measured from the immersion of disk-shaped specimens into deionized distilled water for 24 hours, 1, 2, 4, and 10 weeks. For the fluoride uptake and artificial caries test, standardized restorations were placed along the cementoenamel junction of extracted human molars. Secondary ion mass spectroscopy was used to determine the depth of fluoride uptake into the adjacent axial dentin from the restoration after 1 and 10 weeks. For the artificial caries test, the teeth were immersed into an acidified gelatin gel, pH 4.0, for 10 weeks. The development of recurrent decay was assessed using polarizing light microscopy. Statistical analyses were conducted using ANOVA and Fishers' LSD test (P < or = 0.05). There was generally greater fluoride release and uptake from the conventional glass ionomers, equivalent or less from the resin-modified glass ionomers, and none from the resin composite restorations. The use of an acid conditioner and primer from a dentin bonding system significantly increased the depth of fluoride uptake at 1 week. The additional use of an intermediary adhesive resin layer, however, significantly decreased the depth of fluoride uptake. The maximum depth of fluoride uptake into dentin was 300 microns at 10 weeks. Both conventional and resin-modified glass-ionomer restorations imparted resistance to dentin against the development of recurrent wall carious lesions in vitro. This was attributed to material fluoride release and uptake. PMID- 9227124 TI - Marginal adaptation of composite restorations versus hybrid ionomer/composite sandwich restorations. AB - The aim of this in vitro study was to compare the approximal marginal quality of composite fillings using a dentin bonding system to the marginal quality of hybrid ionomer/composite sandwich restorations. Forty-eight standardized class 2 cavity preparations were prepared in caries-free, human third molars. Twelve preparations at a time were filled either with a composite using the matching dentin bonding system, SZ (Scotchbond MP/Z100) or PP (PROBOND/Prisma TPH) or with hybrid ionomer/composite sandwich fillings VZ (Vitremer/Z100) or DP (Dyract/Prisma TPH). Margins were evaluated before and after thermomechanical loading (TCML) (5000 cycles [+5 degrees C/+55 degrees C], 72.5 N [1,7 Hz]) by quantitative scanning electron microscope analysis using an image analyzing system. Furthermore, microleakage was assessed by dye penetration before and after TCML. Statistical analysis was performed using the Mann-Whitney test at the 0.05 level of significance. SEM analysis after TCML showed significantly fewer marginal gaps at the material/dentin interface with VZ (2.2%), DP (7.3%), and PP (6.0%) compared to SZ (29.6%). After TCML, SZ showed significantly more marginal gaps at the material/dentin interface, whereas VZ, DP, and PP were not susceptible to TCML. PP showed the highest percentage in marginal swelling before (18.2%) and after TCML (15.9%), while VZ showed no marginal swelling at all. VZ showed significantly fewer marginal gaps at the composite/hybrid ionomer interface than DP. After TCML there was no significant difference in marginal gaps between the hybrid ionomer/enamel and the hybrid ionomer/dentin interface for both Vitremer and Dyract. Using the sandwich technique the Z100/enamel interface had significantly more marginal gaps than all other composite/enamel interfaces after TCML. There was no significant difference in microleakage between the test groups at the material/dentin interface. SEM analysis and dye penetration showed that hybrid ionomer/composite sandwich restorations have good marginal qualities and may be an alternative to composite restorations using a dentin bonding system. PMID- 9227123 TI - Diffusion behavior of eugenol from zinc oxide-eugenol mixtures through human and bovine dentin in vitro. AB - The objective of this in vitro study was to evaluate the long-term diffusion behavior of eugenol from different zinc oxide-eugenol mixtures through thin dentin layers. In 24 freshly extracted, caries-free human teeth standardized cavities were prepared exhibiting a dentin surface area of 7 mm2. The remaining thickness of the dentin close to the pulp chamber amounted to at least 0.2 mm. In a second experiment standardized dentin disks (0.2 mm thick) were prepared from 24 bovine incisors. The disks were embedded in epoxy resin, thus exposing a resin free, standardized area of 7 mm2. Specimens from both experiments were inserted into a screw cap of a vial. Cp-Cap, Temp Bond, and two zinc oxide-eugenol mixtures (P/L ratios of 10:1 and 2:1) were applied to each of six specimens from both experiments. The opposing surface was in contact with 1 ml of Ringer solution. After 1 day, 7 days, and 21 days respectively, the amount of penetrated eugenol was determined with High-Performance Liquid Chromatography (HPLC). In both experiments we found a persistent release and diffusion of eugenol, irrespective of the applied material. However, for the second experiment, we saw more pronounced diffusion rates. Furthermore, the materials showed significant differences, particularly for low versus high P/L ratios. PMID- 9227125 TI - Contouring, finishing, and polishing Class 5 restorative materials. AB - Three materials used for class 5 restorations (composite resin, glass ionomer, and resin-modified glass ionomer) were contoured, finished, and polished using seven methods: Matrix only, Sof-Lex disks, Composite Finishing System, Enhance Finishing/Polishing System. Two Striper MFS/MPS System. Two Striper MPS polish only, and contouring burs only. Results were quantified by surface roughness and gloss (reflectance). The glass-ionomer material was rougher and less reflective than composite resin; the smoothness and gloss of the resin-modified glass ionomer were between these two extremes. The original matrix smoothness and gloss could not be reproduced with any of the contour/finish/polish techniques tested on any of the three materials. The smoothest surfaces were achieved using a sequential series of rotary abrasive instruments. PMID- 9227126 TI - 4-META polymethyl methacrylate shear bond strength to titanium. AB - Machined titanium substructures for fixed, complete arch prostheses have been used with various surface pretreatments to enhance acrylic bonding. Since traditional mechanical methods for denture base material retention cannot be used, silane and surface abrasives have been used to enhance retention. The purpose of this study was to determine if a metal bonding denture base material provided a favorable bond strength to machined titanium. Fifteen rod-shaped samples of commercially pure Grade 2 titanium were divided into two groups, five samples representing the control group and 10 samples for the experimental group. The rod samples for the control received no pretreatment, while the experimental samples were pretreated with 110 microns alumina abrasive. Meta-Dent heat-cured polymethyl methacrylate was processed around each sample in a secondary cylindrical shape approximately 0.9 x 1.0 centimeters with the titanium rods extending from each end. A Shell-Nielsen shear test was then performed at a crosshead speed of 0.5 millimeters/minute to determine the bond strength. The experimental group demonstrated a mean bond strength 3.7 times greater than the control group (significant, P = 0.0001, ANOVA). Pretreatment with 110 microns alumina air abrasive significantly enhanced the bond strength of heat-cured metal bonding polymethyl methacrylate to titanium. PMID- 9227127 TI - Amalgam-reinforced stainless steel crown for restoration of permanent molars. AB - This article presents a technique utilizing a stainless steel crown to fabricate a semipermanent provisional restoration of a permanent molar where a diminished risk of occlusal perforation is desired. PMID- 9227128 TI - Bone-resorptive cytokine gene expression in periapical lesions in the rat. AB - Periapical bone destruction is an important pathogenic sequela of pulpal infection. Recent findings from this laboratory have demonstrated that most bone resorbing activity in extracts of rat periapical lesions can be neutralized by an anti-interleukin (IL)-1 alpha antiserum. To further clarify pathogenic mechanisms, bone-resorptive cytokine messenger RNA (mRNA) expression was analyzed in developing rat periapical lesions. The molar teeth of 20 Sprague-Dawley rats were surgically exposed and left open to permit infection from the oral environment. Total cell RNA was isolated from periapical granuloma tissue obtained on days 3, 7, 15 and 30 after exposure. mRNA for IL-1 alpha, IL-1 beta and tumor necrosis factor alpha (TNF-alpha) was amplified by reverse transcription polymerase chain reaction, and levels were approximated by comparison to the parallel amplification of the housekeeping gene glyceraldehyde phosphate dehydrogenase. IL-1 alpha and TNF-alpha mRNA were both highly expressed beginning on day 7, increased on day 15, and declined somewhat on day 30. In contrast, IL-1 beta mRNA was expressed at much lower levels, but with similar kinetics. The kinetics of steady state IL-1 alpha and TNF-alpha mRNA levels were confirmed using the quantitative RNase protection assay, whereas IL-1 beta mRNA could not be detected by this technique. IL-1 alpha mRNA-expressing cells were identified using in situ hybridization and included infiltrating macrophages, as well as resident fibroblasts, endothelial cells and osteoclasts. These results demonstrate that the IL-1 alpha and TNF-alpha genes are highly expressed in developing periapical lesions in the rat and confirm previous studies at the protein level in this model. PMID- 9227129 TI - Effects of acidification on growth and glycolysis of Streptococcus sanguis and Streptococcus mutans. AB - After carbohydrate intake, pH in dental plaque decreases rapidly and reaches about 4 within a few minutes. The acidification not only promotes demineralization of tooth surface but can also cause damage to bacteria in dental plaque. We, therefore, investigated the effect of acidification on the dental plaque bacteria Streptococcus sanguis and Streptococcus mutans. At pH 4.0 and 4.2, both growth and glycolytic activities in these streptococci were repressed. Prolonged acidification (for 60 min at pH 4.0) not only repressed both growth and glycolytic activities but also impaired them in S. sanguis cells with concomitant inactivation of the glycolytic enzymes, hexokinase, phosphofructokinase, glyceraldehydephosphate dehydrogenase and enolase. The impaired abilities of glycolysis and growth recovered following incubation at pH 7.0 for 80-90 min, and this was accompanied by reactivation of the glycolytic enzymes. On the other hand, these impairments were not observed in S. mutans cells exposed to prolonged acidification. These results indicate that the low pH frequently occurring in dental plaque may transiently impair streptococcal glycolysis and growth and that S. mutans is more durable to the acidification than S. sanguis. PMID- 9227130 TI - Effect of sodium and potassium ions on intracellular pH and proton excretion in glycolyzing cells of Streptococcus mutans NCTC 10449 under strictly anaerobic conditions. AB - The effect of sodium and potassium ions on intracellular acid production and acid excretion by glycolyzing cells of Streptococcus mutans was examined. S. mutans NCTC 10449 grown under glucose-limited and strictly anaerobic conditions in a continuous culture system was loaded with bis(carboxyethyl)-carboxyfluorescein, a pH-sensitive fluorescent dye, washed and suspended in 0.00-0.30 M NaCl/KCl solution. The dye allowed for the continuous monitoring of intracellular pH while proton excretion was measured simultaneously with a pH-stat. Sodium ions inhibited and potassium ions, at low pH, accelerated the amount of measurable acid excreted extracellularly. In the presence of both NaCl and KCl, proton excretion following the addition of glucose was slightly higher or similar to that observed in the presence of 0.15 M KCl alone. Sodium and potassium ions did not affect the proton-ATPase enzyme or the intracellular level of ATP, suggesting that these ions did not directly effect proton pumping activity itself. The inhibition of proton excretion by sodium ions was considered to have probably occurred as a result of an indirect inhibition of proton-ATPase activity by the low intracellular pH induced by sodium ions. PMID- 9227131 TI - Molecular analysis of representative Streptococcus gordonii Spp phase variants reveals no differences in the glucosyltransferase structural gene, gtfG. AB - Streptococcus gordonii glucosyltransferase polymerizes sucrose to form glucans, which confer a hard, sucrose-promoted phenotype (Spp+) to colonies on sucrose agar plates. The glucosyltransferase structural gene, gtfG, is positively regulated by the upstream determinant, rgg. Strain Challis undergoes a spontaneous, reversible phase variation between high (Spp+) and low (Spp-) levels of glucosyltransferase activity. Representative strains were examined to gain insights into the basis of glucosyltransferase phase variation. Western blots indicated that the level of glucosyltransferase activity was related to the amount of extracellular glucosyltransferase protein produced by Spp- and Spp+ strains. The nucleotide sequence of rgg and gtfG of the Spp- strain CH97 was found to be identical to that of the Spp+ parent, indicating that DNA differences in these regions are not the basis for glucosyltransferase phase variation. Indeed, 13C-NMR spectroscopy suggested that glucans synthesized by strain CH97 glucosyltransferase were similar to those synthesized by glucosyltransferase of the Spp+ parental strain, indicating a quantitative rather than qualitative change. However, one Spp- strain, CH1C1, had a point mutation in rgg; replacement of the parent rgg with the CH1C1 allele resulted in decreased levels of glucosyltransferase protein and activity. The results indicate that glucosyltransferase phase variation can occur in more than one way, and suggest that glucosyltransferase regulation may involve distally located regulatory gene(s) that affect rgg and/or gtfG expression. PMID- 9227132 TI - Inhibition of purified enolases from oral bacteria by fluoride. AB - Enolase activity in strains of oral streptococci previously has been found to be inhibited by 50% (Ki) by fluoride concentrations ranging from 50 to 300 microM or more in the presence of 0.5 to 1.0 mM inorganic phosphate ions. In this study, enolase was extracted and partly purified by a two-step process from five oral bacterial species and the effect of fluoride on the kinetics of enolase examined. The molecular weight of the putative enolase proteins was 46-48 kDa. The Vmax values ranged from 20 to 323 IU/mg and K(m) for glycerate-2-phosphate from 0.22 to 0.74 mM. Enolase activity was inhibited competitively by fluoride, with Ki values ranging from 16 to 54 microM in the presence of 5 mM inorganic phosphate ions. Ki values for phosphate ranged from 2 to 8 mM. The enolase from Streptococcus sanguis ATCC 10556 was more sensitive to fluoride (Ki = 16 +/- 2) than was enolase from Streptococcus salivarius ATCC 10575 (Ki = 19 +/- 2) or Streptococcus mutans NCTC 10449 (Ki = 40 +/- 4) and all three streptococcal strains were more sensitive to fluoride than either Actinomyces naeslundii WVU 627 (Ki = 46 +/- 6) or Lactobacillus rhamnosus ATCC 7469 (Ki = 54 +/- 6) enolases. The levels of fluoride found to inhibit the streptococcal enolases in this study are much lower than previously reported and are likely to be present in plaque, especially during acidogenesis, and could exert an anti-glycolytic effect. PMID- 9227133 TI - Bacteriocin production and sensitivity among coaggregating and noncoaggregating oral streptococci. AB - Twenty-one oral Streptococcus isolates of known interbacterial coaggregation groups were tested against one another (as both producers and indicators) to detect bacteriocin-like inhibitory activity. In agar-based antagonism tests, seven strains produced small inhibitory zones (< or = 3 mm diameter) but in liquid medium, only strain Streptococcus gordonii DL1 (Challis) produced a detectable antibacterial action (bacteriocin STH1). Five strains were sensitive to bacteriocin STH1, but neither the production of nor the sensitivity to any of the antagonistic agents correlated with coaggregation groupings. Four strains (C219, 903, 118 and Wicky) developed stable resistance in response to the bacteriocin, whereas one isolate (strain 34) remained sensitive following repeated bacteriocin exposure. With one exception (strain 903), bacteriocin STH1 sensitive strains were competent for genetic transformation, but not all competent strains were bacteriocin-sensitive. Bacteriocin-resistant derivatives of transformable strains exhibited decreased competence (80-90% reduction) compared with their parent strains. PMID- 9227134 TI - Detection of identical ribotypes of Porphyromonas gingivalis in patients residing in the United States, Sudan, Romania and Norway. AB - Porphyromonas gingivalis has been isolated from periodontitis lesions in subjects from many geographical locations. The purpose of this investigation was to determine whether similar ribotypes of P. gingivalis could be detected among strains isolated in different countries. A total of 198 isolates of P. gingivalis were obtained from 52 periodontitis patients in Boston (130 isolates), Bergen, Norway (17 isolates), Khartoum, Sudan (26 isolates), and Bucharest, Romania (25 isolates). DNA was isolated from each strain, cut separately by the restriction endonucleases KpnI and PstI. The resulting preparations were subjected to electrophoresis in a 0.8% agarose gel using a Tris-acetate EDTA buffer. Uncut lambda and a 1000-bp fragment of 16S rRNA were included as internal standards in each lane. In addition, a HindIII digest of lambda was present in a separate lane in each run. The DNA fragments were transferred to a nylon membrane by downward capillary transfer. 16S rRNA bands were detected using a 1000-kb digoxigenin labelled probe generated by a polymerase chain reaction. At the same time, a digoxigenin-labelled probe to lambda was employed to detect the internal and molecular weight standards. The bands were detected using antibody to digoxigenin conjugated to alkaline phosphatase and chemiluminescence. The positions of the bands relative to the internal standards were determined and normalized to correct for run-to-run variations, and the molecular weight of each band was determined by comparison with standards within each gel. The resulting data for the 2 enzymes were combined and subjected to cluster analysis using an average unweighted linkage sort. In some instances, isolates that appeared to be of identical ribotype using one endonuclease gave different ribotypes using the other. Strains of P. gingivalis within a subject were usually identical, except for 3 patients who harbored 2 different ribotypes/individual. All subsequent analyses employed a single ribotype strain for each subject. A total of 32 ribotypes were observed for isolates from distant countries. A total of 11.5% of the patients had isolates exhibiting the same ribotype: ribotype 7a. Identical ribotypes of P. gingivalis can be recovered from subgingival plaque samples of periodontitis patients in different countries. PMID- 9227135 TI - Levels of specific immunoglobulin G to Porphyromonas gingivalis in gingival crevicular fluid are related to site disease status. AB - Titers of immunoglobulin G (IgG) directed against Porphyromonas gingivalis in gingival crevicular fluid of 40 periodontitis patients were measured at three sites in each patient (healthy, gingivitis and periodontitis) by enzyme-linked immunosorbent assay. When paired analyses were performed using Wilcoxon signed rank tests, periodontitis sites were found to have lower median titers than gingivitis sites. Both systemic and locally-produced antibodies contribute to the overall gingival crevicular fluid antibody profile. Albumin, in contrast, is derived only from serum, and thus this protein serves as an indicator of serum contribution to gingival crevicular fluid. Correction was therefore made for systemic input to the gingival crevicular fluid IgG profile by expressing the results per unit of albumin. When this was done, periodontitis sites were also found to have significantly lower antibody levels than gingivitis sites. These findings suggest that a failure of local antibody production or reduction in quantities, by, for example, degradation by bacterial proteases, may contribute to the change from a gingivitis to a periodontitis lesion. PMID- 9227136 TI - Optimized oligonucleotides for the differentiation of Prevotella intermedia and Prevotella nigrescens. AB - The gram-negative, anaerobic bacterium Prevotella intermedia plays an important role in the progression of periodontitis, whereas the etiological role of the closely related but phenotypically indistinguishable species Prevotella nigrescens is controversial. To differentiate between these species properly, 16S rDNA/RNA directed, computer-optimized oligonucleotides were designed and tested with 26 P. intermedia, 26 P. nigrescens and a number of closely and more distantly related strains. The oligonucleotides were used as primers in a polymerase chain reaction and could be demonstrated to be species specific with a detection limit of 50 bacterial cells, which could also be detected when diluted 1:10(5) with different plaque bacteria. In addition, the described oligonucleotides were digoxigenin-labeled at the 3' end and used as DNA probes in a dot blot hybridization assay. This assay, although slightly less sensitive than the polymerase chain reaction-based method, gave species-specific reactions and also allowed (semi-)quantification of bacterial cells in clinical specimens. PMID- 9227137 TI - Visualization of an extracellular mucoid layer of Treponema denticola ATCC 35405 and surface sugar lectin analysis of some Treponema species. AB - Slime layers and capsules are common amongst medically relevant bacteria. We herein report that Treponema denticola, which has been associated with periodontitis, synthesizes or acquires an extracellular polysaccharide layer that we have observed through electron microscopy using the polysaccharide-specific dye Alcian blue and phosphotungstate. We have also visualized this extracellular layer by dark-field microscopy of Alcian blue-stained spirochete cells. A representative strain of each of the oral spirochete species T. denticola, Treponema vincentii and Treponema socranskii were differentiated by concanavalin A, phaseolus, lotus A and arachis lectins in a microtiter plate immunoassay for the detection of surface sugars. PMID- 9227138 TI - In vitro antifungal resistance of oral Candida albicans strains in non-AIDS patients. AB - Some cases of oral candidosis are refractory to antifungal treatment. This might be related to development of resistant Candida strains, but susceptibility testing is not standardized and not routinely available, and information related to this problem is scarce in non-AIDS patients. In this study, the in vitro antifungal resistance of oral Candida albicans strains was evaluated. The strains were obtained from a cohort of 72 HIV-negative patients with oral yeast carriage and clinical complaint. Laboratory identification revealed C. albicans in 93% of cases. None of these oral C. albicans isolates showed in vitro resistance to polyenes, but they showed varying resistance levels to fluorocytosine and azoles. This study confirms the usefulness of standardizing susceptibility testing so that it could be routinely available and of realizing a mycological diagnosis including an antifungigram when oral candidosis is suspected, whenever antifungal treatment with azoles is planned. PMID- 9227139 TI - Biologic and clinical evaluation of Syntac and Variolink systems for cohesive pretreatment of hypersensitivity and definitive cementation. AB - Adhesive resin systems have become increasingly popular as they have been demonstrated to infiltrate conditioned vital dentin to cohesively hybridize the biologically altered substrates for adhesive bonding. This paper is a review of the biologic and clinical use of Syntac (Ivoclar Vivadent) and Variolink (Ivoclar Vivadent) adhesive systems for cohesive hybridization of vital dentin to prevent patient postoperative hypersensitivity and bacterial microleakage following clinical preparation. PMID- 9227140 TI - Provisional restorations using the Provipont system. AB - Pending the fabrication of permanent restorations, temporary restorations are used to maintain the function and provide the necessary information for accurate fabrication of permanent restorations. Therefore, the quality and appropriateness of materials used in temporary restorations are important to the final results. This article introduces a new material system for temporary restorations. A case report is used to illustrate the technique and the discussion. PMID- 9227141 TI - Color communication, laboratory fabrication, and cementation. AB - Once considered to be revolutionary, porcelain veneers are now the foundation of most aesthetic dental practices. Requiring only conservative preparations, porcelain veneers can dramatically change a smile. This article discusses the technique necessary to prepare and place porcelain veneers, using a new pressed ceramic system. Close and thorough cooperation and communication between the clinician and the laboratory are essential throughout the procedure, and this article is coauthored by a dentist and a ceramist. PMID- 9227142 TI - Six years of clinical experience with an all-ceramic system. AB - The search for tooth-colored and metal-free restorations is one of the major challenges in dental research. For several decades, ceramic has been used as a restorative material because of its aesthetics and stability. Unfortunately, the survival rate of most all-ceramic systems seems unsatisfactory; due to the natural brittleness of ceramic, fractures have been the primary reason for the high failure rate. Since 1988, the University of Zurich Dental School, Switzerland, has been working with the IPS Empress all-ceramic system (Ivoclar/Williams, Amherst, NY). This article reports the clinical and research data from approximately 3,000 all-ceramic restorations. PMID- 9227143 TI - Restoration of primary molars with Compoglass. AB - The initial glass-ionomer cements have become well-known restorative materials and have achieved success in a variety of procedures. The newest generation of glass-ionomer cements contain more resin, have higher cohesive strengths, and are light cured, compared with previous generations. Other fluoride-releasing resins with glass-ionomer properties have been developed, and they are called "compomers." One such recently introduced material is discussed in this article. A clinical case report is presented, illustrating the utilization of this material in the restoration of a mandibular second primary molar. PMID- 9227144 TI - Denture aesthetics: the union of natural contour, color, and shape. AB - This article reviews a method for placement of full dentures in completely edentulous maxillary and mandibular arches. The selection of a maxillary central incisor is discussed along with the facial/tooth measurement, incisal papilla as a landmark in the maxilla, the midline, and the positioning of the remaining maxillary anterior teeth. The "S" position of the mandibular incisors, occlusion, and the verti-centric dimension are reviewed. The four basic tints of the denture bases are listed, and precision processing is recommended in fabricating the final dentures from information gained from the provisional restorations. PMID- 9227145 TI - A new attachment system for removable partial dentures. AB - How a removable partial denture (RPD) attaches to its abutment teeth is the most important aspect of partial denture design. The reason is obvious: Many critical requirements for optimal design are dependent on the way the removable segment is related and secured to the abutment teeth. To appreciate the importance of the attachment design, it is necessary to understand several principles of overall RPD design. State-of-the-art RPD design is often the most effective means for achieving long-term maintainable health of the remaining teeth. Even weakened teeth with compromised bone support can, with good design, often be used effectively as abutments for RPDs while benefitting from the removable partial. PMID- 9227146 TI - Versatility and aesthetics of the IPS Empress all-ceramic system. AB - With the reestablishment of function no longer a clinical issue, the aesthetic challenge takes precedence, with patients expecting natural-looking results. Since the late 1980s, a leucite-reinforced pressed-glass ceramic (IPS Empress, Ivoclar Williams, Amherst, NY) has been utilized in the anterior region for full crowns and veneers and in the posterior region for crowns, inlays and onlays. This article presents results achieved with the IPS Empress all-ceramic system, which features a unique combination of strength and natural light transmission qualities that resemble the characteristics of natural teeth. PMID- 9227147 TI - A clinical trial of all-ceramic crown restorations: status, fall 1995. AB - Although porcelain-fused-to-metal (PFM) restorations are the most widely used full-coverage crown restoration systems, their inherent properties make the achievement of natural aesthetic restorations an elusive task. In contrast, the all-ceramic system (IPS Empress, Ivoclar North America, Amherst, NY) offers excellent translucency and vitality, without the opacity associated with PFM restorations. Its vitality is further enhanced by an adhesive resin cementation method that conducts the color of the underlying tooth structure. This article reports the status of a 5-year investigation, initiated in 1992, of the clinical performance of the IPS Empress all-ceramic system and the resin cementation system used with it. PMID- 9227149 TI - An interview with Dr. Ronald Jackson, DDS, on esthetic inlays and onlays. Interview by Dr. Gary Severance. AB - Dr. Ronald Jackson is a graduate of West Virginia University School of Dentistry. After a general practice residency and service with the Navy in London, England, he began private practice in Middleburg, VA, in 1976. His practice emphasizes comprehensive restorative and cosmetic dentistry. In an effort to identify leaders in the field of restorative and cosmetic dentistry for Signature readers, I asked Dr. Jackson to meet with me to discuss his experience with inlays and onlays as well as his efforts to educate patients about the different types of restorations available to them. Dr. Jackson has published many articles and lectures extensively on esthetic inlays and onlays. He has twice been a featured speaker at the annual meeting of the American Academy of Cosmetic Dentistry and is scheduled to present at the 1995 meetings of the American Academy of Esthetic Dentistry and the American Society for Dental Aesthetics. Dr. Jackson is an accredited member of the AACD and holds visiting faculty appointments to the postgraduate programs in esthetic dentistry at the State University of New York at Buffalo and Baylor University Schools of Dentistry. He is on the editorial boards of three dental publications and consults to a number of dental manufacturers in the area of new product development and clinical testing of materials. PMID- 9227148 TI - Science, art and nature: a case report. AB - The ceramo-metal restoration still forms the backbone of modern restorative dentistry, despite many new systems. This article discusses a unique ceramo-metal system, its advantages, and clinical and technical applications teamwork between the dentist and the technician is emphasized. IPS Classic (Ivoclar Williams) is a ceramic system with several exclusive features. It encompasses Color Visions, a computer-generated shade system, and the IPS Impulse modifier system allows the ceramist unlimited creativity in color development. PMID- 9227150 TI - Over a decade of success with Heliomolar. AB - In developing improved dental materials and application techniques high standards and goals are set by the clinicians and the manufacturers. This article summarizes the prerequisites and desirable qualities required of composite resins for long-term process. A particular composite has demonstrated predictable and consistent long-term results and has established its place in quality dentistry. The applications for this composite resin are outlined in a clinical procedure. Its anterior/posterior universality is recognized. Several pre- and postoperative examples, in function for up to 12 years, are used to illustrate the results. PMID- 9227151 TI - Ceromers. The evolution of dental materials. PMID- 9227152 TI - The next generation in pit and fissure sealants. AB - Although the benefits of pit and fissure sealants are widely recognized, many clinicians may not routinely include their application as part of a preventive oral health care regimen. This reticence may be due to lack of knowledge regarding technique, appropriate patient populations, and additional advantages to sealant use. This article reviews the background, properties and advantages of Helioseal F (Ivoclar Vivadent, Amherst, NY), which is a resin-based, light curing, fluoride-releasing pit and fissure sealant. The author also provides a checklist of advantageous sealant properties and discusses sealant techniques and the patient profile of sealant candidates. PMID- 9227153 TI - Gold restorations of the distal aspect of cuspid teeth. AB - Proper cavity preparation and fabrication techniques for inlay and onlay restorations require particular scrutiny. Gold has been established as the most effective restorative material for this indication, demonstrating the maintenance of long-term function, marginal integrity, interproximal contacts, effective contouring, and soft tissue compatibility. This article examines the slot inlay restoration, the distal hollow-grind preparation--alone or with a pin, and the class III gold foil technique for the restoration of the distal of the cuspids, a procedure that remains technically demanding for the profession. The author outlines efficient methods by which these restorations can be accomplished for the repair of lesion that reside on the distal aspect of cuspid teeth. PMID- 9227154 TI - Enhanced aesthetics using provisionalization. AB - The proper fabrication of provisional restorations is integral to the overall success of restorative dentistry, as these templates are the initial architecture for the completed case. In a clinical setting, restorations may be utilized for an extended period, during which time the clinician and patient must feel confident. Due partially to a new generation of dental materials with unique properties, provisionalization can be predictably achieved. This article reviews the clinical criteria for the fabrication of interim restorations using a light cure/dual phase temporary crown and bridge material (Provipont DC, Ivoclar Vivadent, Amherst, NY) and its effect on the overall success of restorative procedures, including anterior and posterior restorations. PMID- 9227155 TI - Clinical versatility of new compomer restorative technology. AB - The introduction of restorative materials known as compomers is revolutionizing the field of restorative dentistry. This versatile material combines the most positive attributes of both glass ionomers and composite resins, such as continuous fluoride release, excellent adhesion, ease in placement, exceptional shade blending, and high polishability. This article examines the advantages of the compomer (Compoglass, Ivoclar Vivadent, Amherst, NY) in relation to those of other restorative materials, details the indications for its use, and provides a procedural analysis of its application through a comprehensive description of a case report. PMID- 9227157 TI - Efficient finishing and polishing. PMID- 9227156 TI - Cooperative treatment planning in creating IPS Empress SMILES. AB - With the continued increase in patient expectations for appearance-related restorations, the final result may be deficient in aesthetic qualities, such as color, contour, and shape. This is generally due to a breakdown in communication among the patient, dentist, and laboratory technician with regard to basic smile design principles prior to fabrication of the restoration. With the introduction of advanced porcelain systems (IPS Empress System, Ivoclar Williams, Amherst, NY), the clinician and technician can produce restorations that mimic the optical characteristics and color vitality of natural teeth. This article presents methods to improve communication, effectively evaluate smile design principles, and fabricate aesthetic all-ceramic restorations. PMID- 9227158 TI - Tetric: advanced techniques for a universal direct resin system. AB - No other restorative methodology has been as intensively and critically investigated as that of tooth-colored restorative options. As the biologic and physiochemical shortcomings of earlier adhesive and restorative resins systems have been identified and corrected, newer materials have been formulated, and new techniques utilized. The use of dental dam, attainment of proximal contacts, and the importance of the first layer of composite resin are often overlooked. This article presents various solutions in the quest for adhesive and aesthetic excellence in anterior and posterior applications using Tetric* (Ivoclar Vivadent, Amherst, NY), a microhybrid that meets the optimal criteria for direct composite resin restorations. PMID- 9227159 TI - Click on what you need: dentistry and the Internet. PMID- 9227160 TI - Is there a future for extracorporeal photochemotherapy in the treatment of the rheumatological diseases? PMID- 9227161 TI - Dyslipidemia and rheumatoid arthritis. PMID- 9227162 TI - Apparent hip osteoarthritis in a 16 year old girl. PMID- 9227163 TI - Serum immunoglobulins and the risk of rheumatoid arthritis. AB - OBJECTIVE: Rheumatoid arthritis (RA) is associated with several autoantibodies that can precede the clinical disease. The immunoglobulin concentrations in serum samples before illness were studied to learn more about the immunological process before RA. METHODS: A case-control study was nested within a Finnish cohort of 19,072 adults who had neither arthritis nor a history of it at the baseline examination during 1973-1977. By late 1989, 124 had developed RA, of which 89 were positive for rheumatoid factor (RF). Three controls per each incident case were individually matched for sex, age, and municipality. The concentrations of IgG, IgA, and IgM were measured from stored serum samples. RESULTS: Serum IgG before illness was found to be directly proportional to the risk of RF positive RA, and a non-linear association was present between serum IgA and the risk of RF positive RA. These associations were constant between men and women and other subgroups of the study population and not confounded by serum orosomucoid concentration, level of education, smoking, alcohol intake or body mass index. As adjusted for these factors, the odds ratios (95% confidence intervals) of RF positive RA in the lowest, mid, and highest tertiles of IgG distribution were 1.00, 1.55 (0.81, 2.97), and 2.22 (1.16, 4.26), and in the tertiles of IgA 1.00, 2.23 (1.14, 4.36), and 1.78 (0.89, 3.57), respectively. The associations persisted throughout the entire observation period but were most distinct when the period to the onset of clinical RA was > or = 10 years. IgM carried no predictive significance. None of the serum immunoglobulins predicted the development of RF negative RA. CONCLUSIONS: Increased IgG levels may reflect some, at present unknown process in the early events leading to the development of RA, typically occurring > or = 10 years before the onset of clinical disease. PMID- 9227164 TI - Is addition of sodium fluoride to cyclical etidronate beneficial in the treatment of corticosteroid induced osteoporosis? AB - OBJECTIVE: To investigate whether administration of sodium fluoride (NaF) in addition to cyclical etidronate has a positive effect on bone mineral density (BMD) in patients with established osteoporosis during continued treatment with corticosteroids. PATIENTS AND METHODS: 47 patients who were receiving treatment with corticosteroids were included in a two year randomised, double blind, placebo controlled trial. Established osteoporosis was defined as a history of a peripheral fracture or a vertebral deformity, or both, on a radiograph. All patients were treated with cyclical etidronate, calcium, and either NaF (25 twice daily) or placebo. Vitamin D was supplemented in the case of a low serum 25 (OH) vitamin D concentration. BMD of the lumbar spine and hips was measured at baseline and at 6, 12, 18, and 24 months. RESULTS: After two years of treatment, the BMD of the lumbar spine in the etidronate/NaF group had increased by +9.3% (95% confidence intervals (CI): +2.3% to +16.2%, p < 0.01), while the BMD in the etidronate/placebo group was unchanged: +0.3% (95% CI: -2.2% to +2.8%). The difference in the change in BMD between groups was +8.9% (95% CI: +1.9% to +16.0%, p < 0.01). For the hips, no significant changes in BMD were observed in the etidronate/NaF group after two years: -2.5% (95% CI: -6.8% to +1.8%); in the etidronate/placebo group BMD had significantly decreased: -4.0% (95% CI: -6.6% to -1.4%; p < 0.01). The difference between the groups was not significant: +1.5% (95% CI: -3.4% to +6.4%). No significant differences in number of vertebral deformities and peripheral fractures were observed between the two groups. CONCLUSION: The effect of combination treatment with NaF and etidronate on the BMD of the lumbar spine in corticosteroid treated patients with established osteoporosis is superior to that of etidronate alone. PMID- 9227165 TI - Granulocyte-macrophage colony stimulating factor exacerbates collagen induced arthritis in mice. AB - OBJECTIVE: To examine the effect of granulocyte-macrophage colony stimulating factor (GM-CSF) on disease progression in the collagen induced arthritis (CIA) model in mice. METHODS: DBA/1 mice were primed for a suboptimal CIA response by intradermal injection of chick type II collagen without a secondary immunisation. Three weeks after immunisation the mice were given four to five consecutive daily intraperitoneal injections of recombinant murine GM-CSF (15 micrograms; 5 x 10(5) U), or vehicle, and arthritis development was monitored by clinical scoring of paws and calliper measurements of footpad swelling. At approximately six to eight weeks after immunisation mice were killed, their limbs removed and processed for histological analyses of joint pathology. RESULTS: Control animals receiving a single immunisation with collagen exhibited a varied CIA response both in terms of incidence and severity. Mice treated with GM-CSF at 20 to 25 days after immunisation with collagen had a consistently greater incidence and more rapid onset of disease than the vehicle treated control mice, based on clinical assessment. GM-CSF treated mice showed higher average clinical scores and greater paw swelling than controls. Histological analyses of joints reflected the clinical scores with GM-CSF treated mice displaying more pronounced pathology (synovitis, pannus formation, cartilage and bone damage) than control mice. CONCLUSION: GM-CSF is a potent accelerator of the pathological events leading to chronic inflammatory polyarthritis in murine CIA supporting the notion that GM CSF may play a part in inflammatory polyarthritis, such as rheumatoid arthritis. PMID- 9227166 TI - Technetium-99m labelled liposomes to image experimental arthritis. AB - OBJECTIVES: Liposomes sterically stabilised with polyethylene glycol (PEG) labelled with technetium-99m were tested for their ability to image adjuvant arthritis in a rat model. METHODS: Adjuvant arthritis was induced in the ankle joint of the left hind foot by injection of Mycobacterium butyricum in Freund's incomplete adjuvant in the foot pad. Seven days later animals received the following radiopharmaceuticals labelled with 99mTc (a) non-PEG-liposomes, (b) PEG liposomes or (c) non-specific human polyclonal IgG. For each of the radiopharmaceuticals the in vivo distribution of the radiolabel was monitored both scintigraphically as well as by counting the dissected tissues at two, eight, and 24 hours after injection. RESULTS: The pharmacokinetics of the radiopharmaceuticals differed considerably (halflife in the blood: PEG-liposomes (18 hours) > 99mTc-IgG (3 hours) > non-PEG liposomes (1 hour)). The inflamed focus was visualised with each of the agents. The uptake of each of the radiopharmaceuticals in the inflamed ankle region correlated with their residence time in the blood (inflamed joint uptake: PEG liposomes (1.15% injected dose (ID)/ g) > 99mTc-IgG (0.35% ID/g) > non-PEG-liposomes (0.05% ID/g)). Quantitative analysis of the images showed that the inflamed ankle to background ratio was highest with the PEG-liposomes (7.5 at 24 hours after injection), while with the other two agents this ratio did not exceed 4. CONCLUSION: This study shows that 99mTc-labelled PEG-liposomes may be an excellent agent to visualise arthritis. Increased label uptake in the inflamed joint and increased target to background ratios can be obtained with PEG-liposomes because of their long circulating properties. In addition to their use as vehicles for scintigraphic imaging of arthritis PEG-liposomes might also be used for the site specific delivery of antirheumatic drugs. PMID- 9227167 TI - Effect of disease modifying agents on the lipid profiles of patients with rheumatoid arthritis. AB - OBJECTIVE: To determine the effect of intramuscular gold and oral hydroxychloroquine (HCQ) on the lipid profile of patients with rheumatoid arthritis (RA). METHOD: A prospective randomised clinical trial of 12 months' duration was performed in 100 RA patients. Data on clinical and laboratory parameters of disease activity, and fasting serum lipid samples was collected at baseline and at three monthly intervals over one year. RESULTS: The expected second line response was seen with no significant difference in efficacy between the groups at 12 months. The HCQ group had a significant overall improvement in their lipid profile while there was a trend for lipid profiles in the gold group to worsen. CONCLUSIONS: HCQ is an effective second line agent that has beneficial effects on serum lipids. This should be taken into account when choosing a disease modifying anti-rheumatic drug in patients who suffer from RA and who have significant cardiovascular risk factors. PMID- 9227168 TI - Quantitative magnetic resonance imaging of the knee: a method of measuring response to intra-articular treatments. AB - OBJECTIVES: To investigate the potential of quantitative magnetic resonance imaging (MRI) to differentiate between therapeutically induced changes in inflammation and synovial proliferation in rheumatoid arthritis (RA) of the knee. METHODS: MRI of the knee was performed on patients with RA before and one week after injection with corticosteroid (triamcinolone acetonide, TA group, n = 9) and before, four, and 12 weeks after injection with yttrium-90 plus TA (TA+Y group, n = 7). MRI scans were analysed by subjective visual grading by a trained observer and by computer aided quantitation for three features: synovial fluid volume, synovial pannus volume, and synovial enhancement after intravenous contrast agent. RESULTS: All TA subjects improved clinically at one week but the effects of TA+Y were more variable. TA significantly reduced synovial enhancement and effusion volume, whereas TA+Y at 12 weeks tended to increase synovial enhancement and decrease pannus volume. Quantitative MRI values agreed well with subjective assessment of scans. Comparison of calculated change on MRI scan before and immediately after aspiration with actual volume aspirated showed high correlation (r = 0.96). CONCLUSIONS: Quantitative MRI correlates with subjective visual assessment and, at least for synovial fluid, is accurate. MRI can differentiate actions of two therapeutic modalities on various pathological processes and is sensitive enough to detect change after one week. With the additional advantage of lack of observer bias, it will probably become a useful tool in the development and assessment of existing and novel treatments. PMID- 9227170 TI - Urinary albumin excretion in patients with systemic lupus erythematosus without renal disease. AB - OBJECTIVES: To investigate the prevalence of microalbuminuria, urinary albumin excretion (UAE) between 20-200 micrograms/min, in systemic lupus erythematosus (SLE) patients without clinical renal disease, and to discover if this could predict the development of renal disease. METHODS: This study made six monthly measurements of UAE, creatinine clearance, serological and clinical data in 22 ambulatory women patients with SLE, without clinical renal disease, hypertension, diabetes or heart failure. The patients were followed up for a period of 18 months (four measurements). Age and sex matched healthy controls were used as a comparative group. UAE was measured by nephelometry in three timed overnight urine samples at each visit. RESULTS: There were no significant differences in the creatinine clearance between the control group and the SLE patients. Creatinine clearance did not show significant changes throughout the study period. SLE patients had wide variations in the UAE rate compared with healthy controls. In five patients (5 of 22; 23%), on occasions, there was mild, transient increase in UAE reaching the level of microalbuminuria. During follow up, one patient with basal (4.67 micrograms/min) and six month (4.73 micrograms/min) normal UAE rate, was admitted with a nephrotic syndrome confirmed on biopsy examination to be proliferative lupus nephritis. Six months after beginning treatment with prednisone and cyclophosphamide her UAE rate returned to normal values (4.65 micrograms/min). CONCLUSION: SLE patients without clinical renal disease may have microalbuminuria, although this does not seem to warrant any specific action. PMID- 9227169 TI - Methotrexate in patients with moderate systemic lupus erythematosus (exclusion of renal and central nervous system disease). AB - OBJECTIVES: Methotrexate (MTX) has been used in several autoimmune diseases. Apart from its use in rheumatoid arthritis, MTX has been assessed in small studies in patients with vasculitis, uveitis, and inflammatory bowel disease. The aim of this study was to evaluate the efficacy of MTX in a particular group of patients with systemic lupus erythematosus (SLE). PATIENTS: In an open prospective study 22 patients fulfilling the ACR criteria for SLE were included. Patients had one or more of the following manifestations; active non-destructive polyarthritis, dermatitis, vasculitis of the skin, pleuritis. All patients had been treated with corticosteroids for at least six months without achieving remission. Sixteen patients were taking antimalarial drugs in addition to corticosteroids, which were stopped at the beginning of the trial. Patients with renal and central nervous involvement were excluded from the study. All patients received MTX orally at a dose of 15 mg/week over six months. Corticosteroids were continued. As additional medication only indomethacin up to 100 mg/day was permitted if used before the start of the study. The outcome was evaluated using the SLE disease activity index (SLEDAI). RESULTS: Disease activity was evaluated after six months of MTX treatment. All patients completed the study period. The SLEDAI decreased significantly from mean (SD) 12.2 (3.99) to 4 (3.75) (p = 0.001). The prednisolone dose was reduced from a mean (SD) of 17.4 (12.8) at the beginning to 8.8 (5.36) mg/day at the end point of the study (p = 0.01). MTX was well tolerated. Four patients complained of general malaise. Two patients had transient increases in liver enzymes. In no case did MTX have to be stopped. CONCLUSIONS: In an open prospective study methotrexate was used in SLE patients with particular clinical characteristics. MTX was shown to be effective in reducing disease activity and sparing the dose of corticosteroids. Further controlled studies are necessary. PMID- 9227171 TI - Immunohistochemistry of minor salivary gland biopsy specimens from patients with Sjogren's syndrome with and without hepatitis C virus infection. AB - OBJECTIVES: To characterise phenotypically the minor salivary glands of patients with clinical and histological features of Sjogren's syndrome (SS) infected with hepatitis C virus (HCV). PATIENTS AND METHODS: 75 consecutive patients with SS (31 primary SS, 44 secondary SS) diagnosed by preliminary European classification criteria. The presence of anti-HCV antibodies was detected by commercial third generation ELISA and by a second generation immunoblot assay. Presence of HCV genome in serum was determined by polymerase chain reaction analysis. Expression of CD3, CD4, CD8, CD20, HLA-DR, and CD25 molecules in lymphocytic and epithelial cells on minor salivary glands was detected by immunohistochemical assays. Expression of interferon gamma and interleukin 4 cytokines was determined by in situ hybridisation. RESULTS: Six of 31 primary SS (19%) and one of 44 secondary SS (2%) serum samples were positive for anti-HCV by ELISA. Three samples were positive, three indeterminate, and one sample corresponding to a secondary SS patient was negative by immunoblot. The three immunoblot positive serum samples were also HCV-RNA positive by PCR assay. The study of lymphocytic cells in the diffuse infiltrate of minor salivary glands showed a predominance of the CD3 lymphocytic population. A predominance of CD4 over CD8 T cells (ratio 2:1) was observed in HCV and non-HCV infected patients. The analysis of the lymphocytic focus showed that the HCV infected patients had a predominance of CD20 positive cells. Activation molecules (CD-25 and HLA-DR) were expressed in HCV and non-HCV infected patients in lymphocytic and epithelial cells, however epithelial cell expression of CD25 was low in HCV infected patients. As expected, a pronounced Th1 response was observed in the lymphocytic foci of HCV patients. CONCLUSIONS: HCV infected patients may develop an autoimmune sialadenitis, similar to that described in primary SS. PMID- 9227172 TI - Pericardial involvement in systemic sclerosis. AB - OBJECTIVE: To determine the frequency and histological characteristics of pericardial involvement in systemic sclerosis. METHOD: Necropsy sections of pericardium from 44 patients with systemic sclerosis were studied, together with sections from 19 age/sex matched controls. Sections were stained with haematoxylin and eosin, acid toluidine blue, and elastic van Gieson. Mast cells were counted in 10 random high power fields and the degree of fibrosis was quantified using a Chalkley count. RESULTS: Chronic pericarditis was seen in 31 (77.5%) of the systemic sclerosis cases, but in only one of the controls. The characteristic changes of uraemic pericarditis were not seen. The degree of fibrosis was greater in those with systemic sclerosis, though numbers of mast cells, thought to be important in fibrogenesis, were similar in both groups. Myocardial fibrosis was seen in 15 (37.5%) of systemic sclerosis cases but in none of the controls. CONCLUSION: The incidence of pericarditis and myocardial fibrosis is much greater than in controls. The results indicate that pericarditis is a primary disease (rather than secondary to uraemia). PMID- 9227173 TI - Pain in the rheumatic diseases. PMID- 9227174 TI - Antiperinuclear factors are present in polyarthritis before ACR criteria for rheumatoid arthritis are fulfilled. PMID- 9227175 TI - rhG-CSF resistant neutropenia in SLE. PMID- 9227176 TI - An assessment clinic for routine paediatric otolaryngological surgery. AB - In the current climate of medical practice clinicians are under constant pressure to come up with new ways of increasing efficiency and throughput whilst maintaining a high standard of patient care. Assessment clinics are not a new idea and are widely practised in the National Health Service. The novel feature of the clinic we are describing is that all relevant professionals are present: consultant otolaryngologist, audiologist, paediatric nurse, playleader and preassessment nurse. The Wexham Park Paediatric Otolaryngology Preassessment Clinic has now been running for six years, and in this time we have experienced a marked decrease in both the waiting times for surgery and the non-attendance rate. PMID- 9227177 TI - Whither surgery? PMID- 9227178 TI - Surgery on the Internet. Part 2: The World Wide Web. PMID- 9227179 TI - The British Association of Head and Neck Oncologists within the wider perspectives of the USA and Europe. AB - An account is given of the origin and development of the British Association of Head and Neck Oncologists in relation to American and European Societies. Disillusion with the results of treatment and the poor management available for these patients before 1950, led to the formation of head and neck oncologic societies, initially in the USA, to bring about much needed improvements. Similar views were held in the UK and Western Europe which resulted in the gradual establishment of national societies in the 1960s-80s throughout Western Europe and many other countries. It is believed that these societies should continue to develop and influence medical and planning authorities for the benefit of patients diagnosed as having these uncommon, serious, but often curable tumours. PMID- 9227180 TI - The fellowship and the definition of surgery. PMID- 9227181 TI - Lipids: from intractable grease to oil for the wheels of life. PMID- 9227182 TI - Characteristics of the low-energy reporters in a longitudinal national dietary survey. AB - The aim of the present study was to establish whether the characteristics of members of a large national birth cohort study who submitted diet diaries with implausibly low-energy intake differed from those whose recorded energy intake was more plausible. Survey members (n 1898) recorded their diets in a 7 d diary in household measures. Those whose reported energy intake (EI) as a fraction of their estimated BMR was less than 1.10, here termed low-energy reporters (LER) but often called under-reporters, constituted 20.6% of the study population. None of the variables describing dietary, smoking or exercise behaviour bore a significant relationship with low EI/BMR (< 1.10), neither did those describing region of residence, subjective adequacy of income, current social class, social relations or the social environment of the subjects. Results of logistic regression analysis showed that the only independently significant characteristic for men was higher BMI. In women, in addition to higher BMI, having been overweight or obese as an adult independently, but less significantly, predicted low EI/BMR, while membership as a child of social class III (non-manual), having more children in the household and having a paid job marginally but independently decreased the probability of reporting low EI/BMR. Submission of a diary with EI/BMR < 1.10 7 years earlier in the same survey was an even more powerful predictor of current low EI/BMR than higher BMI in both sexes. The average reported diet-composition of LER was more micronutrient- and protein-rich than that of the others, indicating different dietary, or diet-recording, behaviour in this group of subjects. LER are not a random sample of the survey population, and their characteristics, definable to some extent, put them at risk for lower health status. Although EI/BMR cut-off points can be used to identify LER, the problem of how to use their data is still unresolved. PMID- 9227184 TI - Dynamics of conjunctival impression cytologic changes after vitamin A supplementation. AB - To investigate the chronological changes in conjunctival epithelium after supplementation with a massive oral dose of vitamin A, conjunctival impression cytology (CIC) with transfer was carried out repeatedly among 200 children aged 6 120 months in a randomized, double-blind, placebo-controlled study in Chandigarh (India). Significant conversion to normal CIC started 71-80 d after vitamin A supplementation and by 101-110 d conversion had taken place in all children. Compared with the placebo group, plasma retinol concentration at 100 d post supplement was found to be significantly higher in the vitamin A-supplemented group P = 0.04. This study demonstrates that CIC responds to a massive oral dose of vitamin A 3-4 months after supplementation. These findings should guide future studies and evaluations in which CIC is used to assess response to vitamin A interventions. PMID- 9227183 TI - The effect of oral contraceptive agents on the basal metabolic rate of young women. AB - The use of oral contraceptive agents by women may be a factor that contributes to the observed inter-individual variability in the BMR. We, therefore, measured the BMR, body build and composition in two groups of young women and also assessed their self-reported level of physical activity. One group had been using oral contraceptive agents for a period of 6 months or more (OCA, n 24), while the other group had never used oral contraceptives (NOCA, n 22). There were no significant differences in age, body build or composition. The absolute BMR in the groups were not significantly different when compared using an unpaired t test (OCA: 5841 (SD 471) v. NOCA: 5633 (SD 615) kJ/d). However, using an analysis of covariance, with either body weight or a combination of fat and fat free mass as covariates, the OCA group had a BMR almost 5% higher than that of the NOCA group (OCA: 5871 v. NOCA: 5601 kJ/d; P = 0.002). When those subjects with high self-reported levels of physical activity were excluded, the difference in BMR between the two groups persisted (P = 0.001). An ANOVA of oral contraceptives use and phase of menstrual cycle showed significant differences in BMR with use of oral contraceptives (P = 0.004) but no difference in BMR between phases of the menstrual cycle. In conclusion, the use of oral contraceptive agents deserves consideration when conducting and analysing data from studies on energy metabolism in young women, as it results in a significantly higher BMR. PMID- 9227186 TI - The effect of a high dose of 3-hydroxy-3-methylbutyrate on protein metabolism in growing lambs. AB - The effect of a high dose of 3-hydroxy-3-methylbutyrate (HMB, a leucine catabolite) on protein metabolism was investigated in growing male lambs fed on hay and concentrate. Concentrate was supplemented with either Ca(HMB)2 (4 g/kg) or Ca(CO3)2 in experimental (HMB) and control groups respectively. Both groups consisted of six 2-month old lambs. Three complementary methods to study protein metabolism were carried out consecutively 2.5 months after beginning the dietary treatment: whole body phenylalanine fluxes, postprandial plasma free amino acid time course and fractional rates of protein synthesis in skeletal muscles. Feeding a high dose of HMB led to a significant increase in some plasma free amino acids compared with controls. Total, oxidative and non-oxidative phenylalanine fluxes were not modified by dietary HMB supplementation. Similarly, an acute infusion of HMB, in the control group, did not change these fluxes. In skeletal muscles, fractional rates of protein synthesis were not affected by long term dietary supplementation with HMB. Taken together our results showed that administration of a high dose of HMB to lambs was able to modify plasma free amino acid pattern without any effect on whole-body protein turnover and skeletal muscle protein synthesis. PMID- 9227185 TI - Maternal manipulation of brown adipose tissue and liver development in the ovine fetus during late gestation. AB - We examined the effect of maternal chronic cold exposure, induced by winter shearing ewes 4 weeks before their predicted lambing date, on brown adipose tissue (BAT) and liver development in lambs. Fetuses were sampled from under-fed (60% of energy requirements for maintenance and pregnancy of an unshorn ewe) shorn or unshorn ewes at 126, 140 and 145 d of gestation. Lambs were sampled from ewes within 2 h of birth. Throughout gestation fetal body, BAT and liver weights were similar in shorn and unshorn groups. The level of GDP binding to mitochondrial uncoupling protein remained low throughout gestation, but increased dramatically after birth. Lambs born to shorn ewes possessed more mitochondrial protein and exhibited a significantly higher total thermogenic activity in BAT. Type I iodothyronine 5' deiodinase (EC 3.8.1.4) activity in BAT peaked at birth, as did hepatic iodothyronine 5' deiodinase activity and was significantly greater in lambs born to under-fed shorn ewes, which exhibited a higher plasma triiodothyronine concentration. Chronic maternal adaptations to prolonged cold exposure appear to enable pregnant ewes to compensate for the negative effects of under-feeding on fetal growth and development. PMID- 9227187 TI - Effect of environmental temperature on muscle protein turnover and heat production in tube-fed broiler chickens. AB - The present experiments were undertaken to investigate the effects of environmental temperatures on growth, abdominal fat content, rate of muscle protein turnover, and heat production in tube-fed intact male broiler chickens. Plasma concentrations of thyroxine (T4), triiodothyronine (T3), and corticosterone (CTC) were also examined. Chicks (15 d old) were kept at different environmental temperatures (16, 19, 22, 25, 28, 31, and 34 degrees) and given the experimental diet (200 g crude protein/ kg, 13.57 MJ/kg metabolizable energy) by tube three times daily throughout the 12 d experimental period. In the hot conditions, except for 34 degrees, body-weight gain was significantly higher than in the cold conditions. Thus, food conversion ratios (food:gain ratios) were lower when the birds were exposed to the hot conditions other than 34 degrees. Likewise, abdominal fat content was significantly increased, and heat production was lower in the groups kept under the hot conditions other than 34 degrees. The rate of skeletal muscle protein turnover and plasma concentration of CTC were decreased when the birds were exposed to hot conditions other than 34 degrees, suggesting a role of CTC in the regulation of muscle protein turnover. Plasma concentrations of T4 and T3 were significantly decreased as environmental temperature increased. These results clearly show that plasma concentrations of thyroid hormones and CTC are associated with accelerated muscle protein turnover and heat production. PMID- 9227188 TI - The relationship between in vitro gas production, in vitro microbial biomass yield and 15N incorporation and its implications for the prediction of voluntary feed intake of roughages. AB - The relationship between in vitro gas production, concomitant in vitro apparent and true DM degradability has been examined in forty-two roughages. The partitioning of truly-degraded substrate between gas volume and microbial biomass yield and 15N incorporation into cells was also investigated. The relevance of this partitioning for the regulation of DM intake (DMI) was examined for fifty four roughages. The results can be summarized as follows. In vitro gas production and in vitro apparent and true degradability are highly correlated (P < 0.0001), r being 0.96 and 0.95 respectively. There is an inverse relationship between in vitro gas production and microbial biomass yield (r--0.67, (P < 0.0001) and also 15N enrichment (P < 0.001) when the variables were related to a given unit of substrate truly degraded. Selecting roughages by in vitro gas production may well be a selection against maximum microbial yield and a combination of in vitro gas volume measurements with a complementary determination of the substrate truly degraded is proposed, to calculate a partitioning factor (PF) reflecting the variation of short-chain fatty acid production per unit substrate degraded. PF is calculated as the ratio, substrate truly degraded: gas produced by it. PF was highly significant (P < 0.0001) in DMI prediction when included in stepwise multiple correlations together with in vitro gas volume variables reflecting the extent and rate of gas production; 11% of the variation in DMI was accounted for by the PF. The total model, including extent and rate of gas production and the PF, accounted for 84% of the variation in DMI. Roughages producing proportionally less gas per unit substrate truly degraded had higher feed intakes. PMID- 9227189 TI - Cholesterol-lowering effect of policosanol on rabbits with hypercholesterolaemia induced by a wheat starch-casein diet. AB - The effect of policosanol, a mixture of high-molecular-weight aliphatic alcohols isolated from sugarcane wax, on casein-induced hypercholesterolaemia in rabbits was studied. When policosanol was administered by the oral route once daily for 30 d (50 mg/kg) the increases in plasma total cholesterol and LDL-cholesterol (LDC-C) were significantly reduced when compared with the control group. The incorporation of 3H2O into sterols in the liver was significantly depressed, suggesting inhibition of hepatic cholesterol biosynthesis. The oral administration of policosanol raised the rate of removal of 125I-labelled LDL from serum. Kinetic parameters calculated following injection of [125I]LDL showed than in casein-fed rabbits, the terminal half-life (t1/2) was significantly decreased after policosanol treatment. The hepatic LDL-binding activity was increased after policosanol administration which suggested that the enhanced clearance was due, at least in part, to increased receptor-mediated uptake of LDL by the liver. Considered together, these results suggest that policosanol can significantly reduce the increase of plasma LDL-C in rabbits fed on a wheat starch-casein diet by reducing cholesterol biosynthesis in the liver. Such an effect could account for the enhancement of LDL catabolism through the receptor mediated pathway. PMID- 9227190 TI - Novel dietary strategy for overcoming the antinutritional effects of soyabean whey of high agglutinin content. AB - A diet-switching experiment, which aimed to improve the utilization of soyabean whey was carried out for 61 d with young rats. Feeding was arranged in such a way that after a few days on the soyabean diet, the rats were switched to a high quality lactalbumin diet for a short period, after which the cycle was repeated several times. The weights of the rats at the end of the soyabean phases were significantly less than those of animals pair-fed on a high-quality diet throughout. However, the test group regained the weight loss after switching to the lactalbumin diet. After three cycles there were no significant differences between the weights of the test rats fed on a poor soyabean diet for over a third of the experiment and those fed on the lactalbumin diet throughout. Feed conversion was always significantly higher with test rats in the lactalbumin period than with continually pair-fed controls. Similarly, faecal N losses were significantly higher for test rats in the soyabean phase, but these differences disappeared after switching to the lactalbumin diet. At the end of the experiment there were no significant differences in body protein or lipids between the groups although the pancreas was significantly heavier while the liver was lighter in soyabean-fed rats. The high destruction of trypsin inhibitors in the gut suggests that they probably had little effect on protein digestion in the gut. In contrast, as selective depletion of the agglutinin from soyabean whey removed the nutritional benefit in the lactalbumin part of the cycle, the improved feed conversion in this period must have been the result mainly of the survival and functionality of soyabean agglutinin and the benefits due to the hyperplastic growth and faster renewal of the gut surface it induced. As processing is unnecessary, this novel method is cheap and can be easily adapted for the use of soyabean whey, regarded as a waste product. PMID- 9227191 TI - Immunocompetence in relation to a heat-processed diet (Maillard reaction) in weanling rats. AB - Diets containing unheated casein (CD; control) or a casein-glucose mixture (CGD) previously heated at 140 degrees for 2 h were fed to two groups of young rats for 21 d. Differences in body weight, feed consumption, thymus, and spleen growth, protein metabolism and in vivo immune response were then determined. For this last experiment, animals were inoculated with sheep erythrocytes (SRBC) on day 15 to provide an immunological challenge. No changes were seen in body weight, feed consumption or feed conversion ratios. Neither were significant differences found in spleen weight, protein content, DNA content, DNase (EC 3.1.4.6) activity or lymphocyte count, suggesting that spleen cell growth remained similar in all the animals studied. The CGD induced marked increases in thymus DNA content whilst the protein:DNA ratio became lower. Spleen RNA content was similar in all rats, but thymus RNA content was 29% lower in the CGD group, although this difference did not reach statistical significance. This fact might be a consequence of the low RNase (EC 2.7.7.16) activity and RNase:RNA ratios in the thymus glands of CGD fed animals. Further, the number of splenic plasma cells secreting anti-SRBC antibodies (direct plaque-forming cells) was significantly decreased in the same group. It might be concluded that both diets are adequate for rat growth and that the differences observed in the thymus of CGD-fed rats may be directed towards preserving tissue function. Nevertheless, the CGD did cause immunological disturbances affecting the humoral immune response. PMID- 9227192 TI - Study of magnesium bioavailability using stable isotopes and the inductively coupled plasma mass spectrometry technique in the rat: single and double labelling approaches. AB - The present work aimed to investigate the feasibility of using stable isotopes and inductively-coupled plasma mass spectrometry (ICP/MS) to study Mg absorption in rats. Male Wistar rats, aged 7 weeks and weighing 180 g, were used. They were fed on a semi-purified diet containing 1070 mg Mg/kg, and had free access to feed and distilled water. In the first experiment, after a 16 d adaptation period, two doses of enriched 25Mg (6 and 12 mg) were administered by oral intubation, faeces and urine were collected daily and blood was sampled. Isotope ratios were determined by ICP/MS. 'True' absorption values, using the faecal isotope data, were 0.63 and 0.56 in rats receiving 6 and 12 mg 25Mg respectively, while apparent absorption was 0.50 for two successive periods of metabolic balance studies. Moreover, the oral isotope administration resulted in a measurable isotopic enrichment in plasma within hours which was still detectable on the third day following the isotope administration. In the second experiment, investigating the double labelling technique, similar rats were dosed simultaneously with 5 mg 26Mg orally (premixed with diet) and 0.29 mg 25Mg intravenously. The calculated Mg true absorption values were very similar when calculated from blood or urine data (0.38) but were lower than that obtained from faecal data (0.50). The possible causes of such an unexpected difference and limits of the double labelling technique for Mg absorption are discussed here. Together these results indicate that although 25Mg and 26Mg isotopes have high natural abundance, the described methodology permits meaningful investigations of Mg bioavailability and metabolism. PMID- 9227193 TI - Disentangling the influence of insulin dependent diabetes mellitus on refraction. PMID- 9227194 TI - Nerve fibre layer thickness measurements derived by scanning laser polarimetry: the jury is out. PMID- 9227195 TI - Folding intraocular lenses: materials and methods. PMID- 9227197 TI - Variation of nerve fibre layer thickness measurements with age and ethnicity by scanning laser polarimetry. AB - AIMS: Scanning laser polarimetry is a new technique allowing quantitative analysis of the retinal nerve fibre layer in vivo. This technique was employed to investigate the variation of the retinal nerve fibre layer thickness in a group of normal subjects of different ages and ethnic groups. METHODS: 150 normal volunteers of different ages and ethnic groups were recruited for this study. Three consecutive 15-degree polarimetric maps were acquired for each subjects. Nerve fibre layer thickness measurements were obtained at 1.5 disc diameters from the optic nerve. Four 90-degree quadrants were identified. RESULTS: The mean nerve fibre layer thickness varied from a minimum of 55.4 microns to a maximum of 105.3 microns, with a mean thickness value of 78.2 (SD 10.6) microns. Superior and inferior quadrants showed a comparatively thicker nerve fibre layer than nasal and temporal quadrants. Retinal nerve fibre layer thickness is inversely correlated with age (p < 0.001). White people showed thicker nerve fibre layers than Afro-Caribbeans (p = 0.002). CONCLUSION: The results indicate a progressive reduction of the nerve fibre layer thickness with increasing age. This may be due to a progressive loss of ganglion axons with age as suggested in postmortem studies. A racial difference in nerve fibre layer thickness is present between whites and Afro-Caribbeans. PMID- 9227198 TI - Blind spot size depends on the optic disc topography: a study using SLO controlled scotometry and the Heidelberg retina tomograph. AB - AIMS: To find out whether the size of the blind spot area, determined by static perimetry, depends on the surface topography of the optic disc and its surrounding area. METHODS: Ten eyes were examined; all had a parapapillary atrophy adjacent to the temporal side of the disc. Microperimetry was performed under direct fundus control using a Rodenstock scanning laser ophthalmoscope. The horizontal meridian of the optic discs was examined in 0.5 degree steps using five stimulus sizes (Goldmann I to V), each with 10 different degrees of brightness. Optic disc topography was measured with the Heidelberg retina tomograph (HRT). RESULTS: Stimuli with a high luminance level (Goldmann IV, 4 dB), presented on the horizontal meridian, were seen up to 0.75 degree centrally (that is, towards the optic disc centre) from the temporal edge of the parapapillary atrophy but up to 1.85 degrees centrally from the nasal optic disc border (p < 0.01). Horizontal HRT section profiles of the optic disc consistently showed prominent nasal disc borders contrasting with a shallow excavation within the temporal parapapillary atrophy. CONCLUSIONS: The size of scotomas depends on the surface topography of the tested area. The prominent nasal part of the optic disc appears less 'blind' than the shallow temporal part, probably because of more intensive light scattering by the prominent nasal part of the disc. These considerations should also apply to other scotomas. PMID- 9227196 TI - Long-term influence of insulin dependent diabetes mellitus on refraction and its components: a population based twin study. AB - AIM: To study whether refraction of the eye, or some of its components is influenced by duration of insulin dependent diabetes mellitus. METHODS: From the young cohort of the population based Danish Twin Register, containing 20,888 twin pairs born between 1953 and 1982, all twin pairs having one or both partners affected with IDDM were searched. Autorefraction, autokeratometry, and ultrasonic biometric measurements were carried out on 45 twin pairs: 16 monozygotic (MZ) twin pairs, 14 dizygotic twin pairs of same sex (DZss), and 15 dizygotic twin pairs of opposite sex (DZos). To obtain an estimate of the influence of duration of diabetes, the intrapair differences in duration of diabetes were correlated with intrapair differences in refraction and each of its components. RESULTS: Refraction was statistically significantly negatively correlated with duration of diabetes in the DZss group, and axial length correspondingly positively correlated. Surprisingly, refraction and axial length in the MZ group, adjusted for confounding factors, were correlated with diabetes duration in the opposite direction than in the DZss group, although not reaching statistical significance. Lens thickness was statistically significantly positively correlated with duration of diabetes in both MZ and DZ twins. Anterior chamber depth was negatively correlated with duration of diabetes in all the zygosity groups. CONCLUSIONS: Studies of relations between refraction and duration of diabetes show diverging results. In the MZ group, a tendency to reduced axial length and corresponding hyperopia with increasing duration of diabetes was found. However, in the DZ group of same sex the opposite tendency was found. Increasing lens thickness and decreasing anterior chamber depth with increasing duration of diabetes have been confirmed in this study. PMID- 9227199 TI - Reproducibility and sensitivity of scanning laser Doppler flowmetry during graded changes in PO2. AB - AIMS/BACKGROUND: Recently a commercially available scanning laser Doppler flowmeter has been produced, which provides two dimensional maps of the retinal perfusion. The aim of the present study was to investigate the reproducibility and the sensitivity of these measurements. METHODS: 16 healthy subjects were randomised to inhale different gas mixtures of oxygen and nitrogen in a double blind crossover study. The following gas mixtures of oxygen and nitrogen were administered: 100% oxygen + 0% nitrogen, 80% oxygen + 20% nitrogen, 60% oxygen + 40% nitrogen, 40% oxygen + 60% nitrogen, 30% oxygen + 70% nitrogen, 20% oxygen + 80% nitrogen, 15% oxygen + 85% nitrogen, and 10% oxygen + 90% nitrogen. Retinal haemodynamic variables and systemic haemodynamics were measured during all inhalation periods. Recordings under resting conditions were performed three times to calculate intraclass coefficients. RESULTS: In two subjects we did not obtain technically adequate results. A dose dependent change in retinal blood flow during graded oxygen breathing was observed (p < 0.001). During 100% oxygen breathing blood flow decrease was between 29% and 33%, whereas blood flow increase was between 28% and 33% during inhalation of 10% oxygen + 90% nitrogen. CONCLUSIONS: Scanning laser Doppler flowmetry has an acceptable reproducibility and is appropriate for description of the effect of graded changes in PO2 on retinal haemodynamics. The main problems with the system are the large zero offset, the fixation during retinal scanning, and the neglect of blood flow changes during the cardiac cycle. PMID- 9227200 TI - Effect of breathing 100% oxygen on retinal and optic nerve head capillary blood flow in smokers and non-smokers. AB - AIM: The effect of breathing 100% oxygen on retinal and optic nerve head capillary blood flow in smokers and non-smokers was investigated using scanning laser Doppler flowmetry (SLDF) as a new non-invasive method to visualise and quantify ocular blood flow. METHOD: 10 eyes of 10 young healthy non-smoking volunteers (mean age 26 (SD 3) years) and nine eyes of nine young healthy smoking volunteers (mean age 26 (4) years) were investigated. All participants were asked not to smoke or consume caffeine containing drinks for at least 4 hours before the measurements. Blood flow measurements were performed before and after 100% oxygen was applied to the subjects through a mask over a period of 5 minutes (6 litres per minute). Juxtapapillary retinal and optic nerve head blood flow were determined in arbitrary units using SLDF representing a combination of laser Doppler flowmetry and a scanning laser system allowing visualisation and quantification of the retinal and optic nerve head blood flow. Blood flow was determined in an area of 100 microns x 100 microns. The level of carboxyhaemoglobin was determined in all subjects. A Wilcoxon matched pairs signed ranks test (non-parametric) was used for statistical evaluation. RESULTS: In the non-smoking group, retinal 'flow' was reduced by 33% (p = 0.005), optic nerve head 'flow' by 37% (p = 0.005). In the smoking group retinal flow was reduced by 10% (p = 0.01), optic nerve head flow by 13% (p < 0.008). The difference in reactivity to oxygen breathing between smokers and non-smokers was highly significant (p < 0.00001). Increased carboxyhaemoglobin levels were not found in either of the groups. A significant reduction of the mean arterial blood pressure of 6% (5%) (p < 0.02) was observed in the non-smoking group after administration of oxygen. CONCLUSION: These results indicate that hyperoxia leads to a decrease in capillary blood flow of the retina and optic nerve head secondary to vasoconstriction, and that smokers do not respond to oxygen breathing as non-smokers do. The findings might be based on factors such as long term effects of nicotine on the sympathetic and parasympathetic nervous system. PMID- 9227201 TI - Effects on IOP restoration and blood-aqueous barrier after long-term treatment with latanoprost in open angle glaucoma and ocular hypertension. AB - AIMS: To evaluate whether long-term treatment with the prostaglandin analogue latanoprost has a deleterious effect on the blood-aqueous barrier (BAB) and to determine the duration of the effect on intraocular pressure (IOP) after withdrawal of treatment. METHODS: Patients with ocular hypertension or glaucoma were topically treated with latanoprost 50 micrograms/ml once daily for 6-12 months. In 26 patients IOP was followed for 14 days after withdrawal of treatment. Aqueous flare was measured with a laser flare meter during 6-12 months' treatment in 16 patients. RESULTS: On the last day of treatment IOP was 6.9 mm Hg (95% CI 5.3-8.5) lower than before treatment. It increased slowly during the follow up period but was still 1.3 mm Hg (95% CI 0.2-2.5) lower than pretreatment IOP 14 days after cessation of treatment. No change in aqueous flare was seen throughout the study. CONCLUSION: Latanoprost has no clinically significant effect on the permeability of the BAB and IOP will return to pretreatment levels within a few weeks, indicating that latanoprost is safe for long-term treatment. PMID- 9227203 TI - Intrafamilial variation of the phenotype in Bardet-Biedl syndrome. AB - AIMS: To describe the variation of the phenotype within families with several individuals with Bardet-Biedl syndrome. METHODS: The phenotypes of affected siblings in 11 Scandinavian families with two or more members who had at least three of the features: retinal dystrophy, polydactyly, obesity, hypogenitalism, and mental retardation, were compared [corrected]. Individuals without retinal dystrophy were excluded. RESULTS: Intrafamilial variation of expressivity of the features obesity, polydactyly, abnormal radiograms of the extremities, hypogenitalism, short stature, paraplegia, and dental abnormalities was found. The retinal dystrophy varied with respect to both the onset of symptoms and the course of the disease. The morphology of the fundus, however, was consistent within the families. The disorder showed statistically significant genetic linkage to the BBS4 locus on chromosome 15 in the affected siblings in two of the families, but the clinical features in these patients did not differ from the other cases of Bardet-Biedl syndrome. CONCLUSION: Comparison of siblings with the Bardet-Biedl syndrome showed variation of the typical features. In addition, the course of retinal dystrophy varied. No distinctive clinical features were found to separate the BBS4 phenotype from the remaining patients. PMID- 9227202 TI - Ocular abnormalities in thin basement membrane disease. AB - AIM/BACKGROUND: Alport syndrome is an X linked disease that results in renal failure, deafness, and ocular abnormalities including a dot and fleck retinopathy and anterior lenticonus. The ultrastructural appearance of the glomerular basement membrane in thin basement membrane disease (TBMD) resembles that seen in some patients with Alport syndrome, and in some cases this disease is inherited too. The aim of this study was to determine whether patients with TBMD have any ocular abnormalities. METHODS: The eyes of 17 unrelated individuals with TBMD were studied by slit-lamp, including biomicroscopic fundus examination with a 78 D lens, by direct ophthalmoscopy, and by fundal photographs. The findings were compared with those in patients with IgA glomerulonephritis or Alport syndrome, and in normals. RESULTS: No patient with TBMD had a dot and fleck retinopathy or anterior lenticonus. A corneal dystrophy (n = 2) or pigmentation (n = 1), and retinal pigment epithelial clumping and maculopathy (n = 1) were noted. Corneal, lens, and retinal dots were found in five (29%), three (18%), and 16 (94%) patients, respectively, but these were also demonstrated in individuals with other renal diseases and in normal individuals. CONCLUSIONS: The dot and fleck retinopathy and anterior lenticonus typical of Alport syndrome do not occur in TBMD. The protein abnormality and genetic defect in TBMD are not known, but the lack of ocular lesions suggests that the abnormal protein in this disease is more sparsely distributed or less important in the basement membranes of the eye than of the kidney. Alternatively, the protein may be less affected by the mutations responsible for TBMD. PMID- 9227204 TI - Orbital metastases: diagnosis and course. AB - AIMS: Three issues were investigated in adult outpatients with orbital metastases. One, how accurate are current diagnostic methods? Two, what is the survival associated with orbital metastases? Three, did any clinical factors correlate with prognosis in this patient cohort? METHODS: Retrospective analysis of patients with orbital metastases managed in an ocular oncology unit. RESULTS: 11 of 31 (35%) patients had no known primary malignancy at the time of orbital diagnosis. In eight of 31 (26%) computed tomography and/or magnetic resonance imaging data did not yield the diagnosis of metastases. In 15 of 17 (88%) cases a fine needle aspiration biopsy was diagnostic. Several types of therapy were used. The median survival was 1.3 years. CONCLUSION: Orbital metastases, even with newer diagnostic techniques can be difficult to diagnose. Management was based on location and extent of both orbital and systemic disease as well as vision. In most cases, orbital symptoms were palliated; however, survival was dismal. No clinical factor correlated with prognosis. PMID- 9227205 TI - Influence of topical human epidermal growth factor on postkeratoplasty re epithelialisation. AB - AIM: To test the efficacy and safety of recombinant human epidermal growth factor (hEGF) on corneal re-epithelialisation following penetrating keratoplasty. METHODS: A prospective, randomised, placebo controlled study was carried out in which patients were matched for diagnosis and received either hEGF ophthalmic solution (30 micrograms/ml or 100 micrograms/ml) or placebo in a double masked fashion. Matched pairs of patients received donor corneas from the same donor and were operated by the same surgeon on the same day. At the end of surgery all donor epithelium was removed mechanically. Patients were examined twice daily and fluorescein stained photographs were taken until the epithelium had closed. The area of the defect was measured by planimetry of the fluorescein stained defect on the photographs. RESULTS: There were no significant differences in re epithelialisation of the donor cornea between the placebo group and the group treated with 30 micrograms/ml hEGF. Time until complete closure was slightly longer with 100 micrograms/ml hEGF compared with 30 micrograms/ml hEGF and with placebo. Mean healing rate of the epithelial defect with 100 micrograms/ml hEGF was significantly slower than in the other groups. CONCLUSION: No significant acceleration of corneal re-epithelialisation was demonstrated with the use of recombinant hEGF after penetrating keratoplasty in humans. PMID- 9227206 TI - Macrophages and MHC class II positive cells in the choroid during endotoxin induced uveitis. AB - AIMS/BACKGROUND: Endotoxin induced uveitis has been regarded as a model for acute anterior uveitis and until now little was known about choroidal involvement. The aim of this study was to investigate changes in macrophages and MHC class II positive cells in the choroid of Lewis rats during endotoxin induced uveitis. METHODS: Choroid-sclera wholemounts were isolated from normal Lewis rats and at different time points--4, 8, 16, 24, 48, 72, and 96 hours, and 7, 10, and 14 days after a footpad injection of 200 micrograms of lipopolysaccharide (LPS). Immunohistochemistry was performed using the monoclonal antibodies ED1 (monocytes, macrophages, dendritic cells), and OX6 (MHC class II antigen). RESULTS: In normal rats, two layers of macrophages were identified in the choroid; a layer located immediately beneath the retinal pigment epithelium (RPE) and a layer bordering the sclera. The density of ED1 positive cells in the layer bordering the RPE cells was 902 (SD 132) cells/mm2 whereas the scleral layer had a cell density of 389 (73) cells/mm2. Based on morphology, positive cells could be divided into two main categories; pleomorphic/round cells and dendritiform cells with varying appearances, with the latter being predominant in normal eyes. A network of MHC class II positive dendritic cells was found in the choroid, beneath the RPE, with a density of 659 (96) cells/mm2. No MHC class II positive cells were found in the macrophage layer bordering the sclera. LPS injection caused a massive influx of ED1 positive macrophages in the area below the RPE cells but did not result in an influx of macrophages at the scleral side of the choroid. The infiltrate reached a maximum at 16 hours following LPS injection and decreased at 96 hours. The morphology of the infiltrating cells was pleomorphic/round at early stages of inflammation and changed into a dendritiform cell population later. The number of MHC class II positive cells on the anterior side of the choroid increased 8 hours after injection and reached a peak at 72-96 hours. MHC class II positive cells were not observed in the vicinity of the sclera at any time after LPS injection. Both resident and MHC class II positive dendritic cell numbers returned to normal values at day 14 following LPS injection. CONCLUSIONS: These results indicate that the choroid is severely inflamed after systemic LPS administration to Lewis rats and suggests that endotoxin induced uveitis may serve as a model for generalised uveitis in humans. PMID- 9227207 TI - Increased number of IgE positive Langerhans cells in the conjunctiva of patients with atopic dermatitis. AB - AIM: To determine the role of Langerhans cells (LCs) found to bear IgE in patients with atopic dermatitis (AD) by evaluating the surface distribution of these cells in the conjunctival epithelium and epidermis of skin lesions in patients with AD. METHODS: The double labelling method was used to evaluate IgE positive cells that were positive for anti-CD1a or anti-CD23 antibody in an epithelial sheet of the conjunctival limbus. Specimens of conjunctiva were obtained from 12 men, six of whom had AD and ocular complications. Five patients without atopic disease served as controls, plus one additional patient with asthma but no AD. A similar study was conducted using epidermal sheets obtained from two patients with AD and from one without AD. RESULTS: The number of CD1a+ cells present in the conjunctival epithelium of the patients with AD significantly exceeded that of the patients without AD. Most CD1a+ cells in the conjunctival epithelium and epidermis from the patients with AD bore IgE on their surfaces. Few such cells from patients without AD bore IgE. No CD23+ cells were found in the patients with or without AD. CONCLUSIONS: The presence of an increased number of LCs bearing IgE on their surfaces in the conjunctival epithelium of patients with AD suggests that these cells may be involved in eliciting the hypersensitivity reaction and participate in ocular inflammation. PMID- 9227208 TI - Intraocular lens durability after a mean of 10.9 years' implantation in humans. AB - AIMS/BACKGROUND: To clarify whether intraocular lens (IOL) implantation in the human eye affects the durability of polymethylmethacrylate over an average period of 10.9 years. METHODS: Shearing stress and extent of damage following neodymium (Nd):YAG laser application to 18 study and 12 control optics were examined. RESULTS: No significant difference was found between the study and the control IOLs in shearing stress and extent of damage following Nd:YAG. CONCLUSION: An average 10.9 years' implantation in humans does not affect either the shearing stress or extent of damage following Nd:YAG shots of polymethylmethacrylate. PMID- 9227210 TI - Recurrent uveitis in a patient with adult onset cyclic neutropenia associated with increased large granular lymphocytes. PMID- 9227209 TI - The genetics of primary open angle glaucoma. PMID- 9227211 TI - Delayed acute retinal necrosis after herpetic encephalitis. PMID- 9227212 TI - Late treatment of methanol blindness. PMID- 9227213 TI - Intracranial plasmacytoma presenting with optic nerve compression. PMID- 9227214 TI - Conjunctival and lacrimal sac pigmentation by kohl (eyeliner) PMID- 9227215 TI - An unusual case of map-dot epithelial dystrophy. PMID- 9227216 TI - Intraocular pressure changes after peribulbar injections with and without ocular compression. PMID- 9227217 TI - Preschool vision screening. PMID- 9227218 TI - First day follow up for routine phacoemulsification? PMID- 9227219 TI - A case of Leber's hereditary optic neuropathy with elevated blood levels of lactate and pyruvate. PMID- 9227220 TI - Accelerated ocular hypertensive response to topical steroids in children. PMID- 9227221 TI - Feasibility of automated visual field examination in children. PMID- 9227222 TI - Revised radiation doses for typical X-ray examinations. Report on a recent review of doses to patients from medical X-ray examinations in the UK by NRPB. National Radiological Protection Board. PMID- 9227223 TI - Computed tomography of the chest in diving-related pulmonary barotrauma. AB - Arterial gas embolism due to barotrauma of the lungs is a severe complication in compressed air diving. Precipitating factors are often missed on plain chest radiology. This study was conducted to detect occult lung disease predisposing to the development of pulmonary barotrauma in conditions associated with a change in ambient pressure. During the past 4 years, 11 patients who have suffered pulmonary barotrauma with or without subsequent development of cerebral or spinal arterial gas embolism underwent computed tomography of the chest several days post-injury. Examinations were conducted either using the conventional technique (n = 7) or, more recently, in the spiral mode (n = 4). Chest radiographs were available in all cases. In five patients CT revealed subpleural emphysematous blebs or cysts that were not detected by conventional radiography. Follow-up studies performed in two of these cases several months post-injury showed that the cystic lesions did not resolve. We assume that the lung cysts or blebs are preexisting conditions which caused pulmonary barotrauma. Computed tomography of the chest, preferably in the spiral mode, is recommended in any case of suspected pulmonary barotrauma in order to evaluate the possibility of pre-existing pathology and to predict future fitness to dive. PMID- 9227224 TI - Dynamic MR imaging of invasive breast cancer: correlation with tumour grade and other histological factors. AB - The purpose of this study was to explore the association between dynamic MR enhancement characteristics and histopathological prognostic factors of invasive breast cancer. 53 women with primary invasive breast cancer underwent dynamic contrast enhanced breast MRI. Region of interest (ROI) analysis was performed on synthetic images obtained by kinetic modelling of the dynamic data. Operator defined, large ROIs and computer-defined, 9-pixel ROIs were selected for each tumour. The relative increase in mean ROI pixel intensity was expressed in the form of enhancement ratios. Univariate and multivariate analyses were performed to explore the association of these ratios with standard histological factors, including tumour size, histopathological classification, histological grade, the presence of extensive in situ component and lymphovascular invasion, multifocal disease, and axillary lymph node status. All enhancement ratios showed significant differences between node-positive and node-negative tumours (max. p = 0.002). However, automated ROI ratios showed less overlap between node-positive and node-negative carcinomas than did large ROI ratios. A strongly significant association was observed between all automated ROI enhancement ratios and histological tumour grade (max. p = 0.001). Based on stepwise multiple regression analysis, node status and histological grade were the only histopathological factors with a significant independent effect on the enhancement characteristics. In summary, there is a strong association between dynamic MR characteristics and two important prognostic markers of invasive breast cancer, namely axillary node status and histological grade. This may allow MRI to be used in pre-operative predictions of tumour behaviour and biological activity. PMID- 9227225 TI - MRI of breast cancer: influence of chemotherapy on sensitivity. AB - MR mammography (MRM) seems to be a sensitive method for detection of breast cancer. The effect of cytotoxic agents on the dynamics of contrast medium uptake in primary breast carcinoma or recurrent disease is not known. This study addresses this question and evaluates MRM as a method of monitoring therapeutic success. A total of 13 patients (age range 34-62 years) with histologically confirmed breast cancer were investigated. The patients received neoadjuvant intravenous (iv) chemotherapy. MRM and interpretation of the dynamic measurements were performed in a standardized manner after positioning the patient in a double breast coil. A gradient echo sequence (Flash 3D, TE 5 ms, TR 12 ms, flip angle 25 degrees) was acquired before and 1, 2, 3 and 8 min after intravenous injection of Gd-DTPA 0.15 mmol kg-1 body weight. A T2 weighted SE sequence (TE 103 ms, TR 6900 ms, 4 mm, field of view 350 mm) was also obtained. MRM was performed prior to histological evaluation and after chemotherapy. All cases of malignancy were correctly diagnosed with MRM. Based on MR findings, eight patients were classified as "responders" and the remaining as "non-responders". In the "responders" a flattening of the Gd-DTPA uptake curve after the first cycle of chemotherapy of complete absence of Gd-DTPA uptake after the fourth cycle was observed. The change in Gd-DTPA uptake behaviour led to an underestimation of the extent of tumour in two patients and false negative findings in four patients. MRM provides information regarding response to therapy following the first cycle. MRM does not provide information regarding invasive tumour tissue in "responders". PMID- 9227226 TI - Technetium-99m sestamibi scintigraphy, magnetic resonance imaging and venous blood sampling in persistent and recurrent hyperparathyroidism. AB - Surgical treatment for primary hyperparathyroidism (HPT) is effective in 90% of cases. Recurrent or persistent HPT occurs in 10% of cases. Parathyroid imaging is indicated to confirm and locate an abnormal gland before reoperation. The aim of this study was to evaluate whether the combination of 99Tcm sestamibi scintigraphy, MRI and venous blood sampling (VBS) improved the overall sensitivity for abnormal parathyroid gland detection. 18 patients with recurrent or persistent HPT underwent sestamibi scintigraphy (n = 18), MRI (T1 weighted and STIR sequences) (n = 18) and venous blood sampling (n = 12) at different sites (internal jugular veins, innominate veins, and superior vena cava). All patients underwent surgical exploration. MRI yielded positive results in 15 cases (sensitivity 88%), sestamibi scintigraphy in 14 cases (83%) and VBS in 10 cases out of 12 (83%). Combined results of MRI, sestamibi and VBS yielded positive results in 16 cases (94%). The combination of MRI, sestamibi scintigraphy and VBS improved accuracy in detecting abnormal parathyroid glands before reoperation. PMID- 9227227 TI - MRI of inner ear anatomy using 3D MP-RAGE and 3D CISS sequences. AB - The aim of this study was to compare contrast enhanced 3D MP-RAGE (magnetization prepared rapid gradient echo), unenhanced 3D MP-RAGE and 3D CISS (constructive interference in steady state) in the evaluation of anatomical detail of the inner ear and facial nerve. 60 persons with no abnormalities and no or non-specific symptoms were examined with MRI. All examinations were performed using a 1.5 T MR unit. The detectability of anatomical details was evaluated by agreement of three radiologists. Statistical evaluation of the results was achieved by the two tailed Wilcoxon's test. In 86-95% of the cases, 3D CISS resulted in excellent visibility of the basal and second turn and apex of the cochlea, the vestibule and semicircular canals, as well as the nerves within the internal auditory canal. There was a significantly better visualization with CISS than with MP RAGE. Detectability of the extrameatal facial nerve was best using contrast enhanced 3D MP-RAGE in 91-96% of the cases (labyrinthine segment 96.7%; geniculate ganglion 95%; tympanic segment 91.7%; vertical segment 95%). The detection of the meatal seventh nerve was best using CISS, whilst unenhanced MP RAGE gave significantly better results than contrast enhanced MP-RAGE. These results suggest that unenhanced and contrast enhanced 3D MP-RAGE and 3D CISS sequences are complementary and not alternative MRI techniques. Both T1 and T2 weighted 3D MR imaging of the temporal bone is of advantage when compared with 2D MR sequences due to improved contrast, geometrical resolution and the possibility of adequate reconstruction of anatomical structures. PMID- 9227228 TI - A comparison between 111In-anti-E-selectin mAb and 99Tcm-labelled human non specific immunoglobulin in radionuclide imaging of rheumatoid arthritis. AB - We have developed and validated a method for imaging inflammation using a monoclonal antibody (1.2B6) against E-selectin, an endothelial-cell specific adhesion molecule. This study was undertaken to compare 111In-1.2B6 with 99Tcm labelled non-specific IgG (99Tcm-HIG) in the detection of synovitis in 11 patients with rheumatoid arthritis (RA). Imaging was performed 4 h and 20-24 h post-injection (pi) of 555 MBq 99Tcm-HIG and 15 MBq 111In-1.2B6. Scintigraphic results were compared with clinical scores of joint involvement. Joint uptake was semiquantitated. The scintigraphic appearances with both tracers correlated well, although 111In-1.2B6 at 24 h showed the highest detection rate. Taking joint tenderness or swelling as evidence of clinical activity, the sensitivity of 111In 1.2B6 at 4 h and 24 h was 69% and 82%, respectively, compared with 69% and 62% for 99Tcm-HIG. 111In-1.2B6 also displayed abnormal activity over a number of joints that appeared silent on clinical examination. Joint-to-soft tissue ratios were higher for 111In-1.2B6 at 24 h (4.0 +/- 1.9; p < 0.0001 vs all) than at 4 h (2.4 +/- 1.4) or than for 99Tcm-HIG at 4 h and 24 h (1.6 +/- 0.5 and 2.3 +/- 0.7, respectively). Net 111In counts over joints increased significantly between 4 h and 24 h (mean change: 54 +/- 40%). This study demonstrates that 111In-1.2B6 scintigraphy is a sensitive method by which to assess RA activity and that targeting is more intense and specific than using 99Tcm-HIG. However, the optimum time for 111In-1.2B6 scintigraphy is 24 h whereas good results are already obtained with 99Tc-HIG at 4 h pi. Current efforts are directed at developing 99Tcm-labelled 1.2B6 for imaging endothelial activation. PMID- 9227229 TI - Influence of number of views and mammographic film density on the detection of invasive cancers: results from the NHS Breast Screening Programme. AB - The National Health Service Breast Screening Programme (NHSBSP) has recommended the adoption of two view mammography at the prevalent screen, and the use of a target film density in the range 1.4-1.8. The aim of this study was to review the impact of number of views and optical density on the detection of invasive cancers. The last four annual returns for screening centres in the NHSBSP have been analysed retrospectively for 2827342 women aged 50-64 years attending their first (prevalent) screening examination. The detection of invasive cancers was assessed in relation to the number of views and film density using the age adjusted, Standardized Detection Ratio measure of screening performance. Typical film densities were reported for each screening year by local physicists, and the average value for all mammography sets at each programme calculated, and found to vary from 0.85 to 1.85. The mean film density across the NHSBSP rose progressively from 1.30 (SD = 0.21) in 1991/2 to 1.57 (SD = 0.12) in 1994/5. Programmes using single view mammography (MLO) and an optical density less than 1.4 detected 76% (95% CI 74-79%) of the expected invasive cancers. Programmes using two view mammography (MLO and CC) and an optical density equal to or greater than 1.4 detected 95% (95% CI 92-98%) of the expected invasive cancers. In 1994/95 when more programmes used the recommended screening modes, the NHSBSP detected 96% (95% CI 92-101%) of the expected invasive cancers at prevalent screening. The detection of invasive cancers was highest where programmes used two views with a film density in the range 1.4-1.8. The results provide evidence of the benefit of the recommended protocol for prevalent screening and indicate that from 1995/96 when all programmes will be using the recommended protocol, it is likely that the detection rates and interval cancer rates from prevalent screens in the NHSBSP will be close to the figures in the Swedish-Two County Trial. PMID- 9227230 TI - Patient dose reduction by audit of grid usage in barium enemas. AB - Patient dose reduction may be achieved by removal of the antiscatter grid during fluoroscopy when fine detail is not required, such as during the filling phase of barium enemas. A questionnaire revealed that none of 22 trainee radiologists routinely removed the grid for the filling phase of adult barium enemas. Following this, 100 consecutive barium enemas were observed, with assessment of patient radiation dose by means of a dose-area product meter. The grid was removed in only six examinations. Phantom measurements implied a possible 46% reduction in dose for the filling phase by correct grid usage. These data were presented at a local clinical audit meeting, resulting in a change in departmental policy to recommend routine removal of the grid for the filling phase of barium enemas. In a follow-up audit the grid was removed in 96 of 100 consecutive enemas, resulting in a significant 46.5% dose reduction for the filling phase (95% confidence interval 38-61%). Further re-audit has demonstrated continued good practice. PMID- 9227231 TI - The impact of cardiology on the collective effective dose in the North of England. AB - Two cardiology X-ray rooms were monitored with dose-area product meters as part of a Regional Patient Dosimetry Programme. Dose-area product measurements on over 2000 patients undergoing examinations in the cardiology rooms are presented. The data have been corrected according to patient size where possible. In room A mean dose-area product values for coronary angiography, coronary angioplasty, radiofrequency ablation and mitral valvuloplasty were found to be 47.7, 72.2, 91.1 and 161.9 Gy cm2 respectively. In room B mean dose-area product values for coronary angiography and coronary angioplasty were found to be 23.4 and 51.6 Gy cm2 respectively. Observational studies were used to deduce the typical projections and technique factors. This typical examination was used to simulate an angiogram from which it was possible to derive factors to convert measured dose-area product values into estimates of effective dose. In room A, the effective doses were estimated to be 9.4, 14.2, 17.3 and 29.3 mSv for coronary angiography, coronary angioplasty, radiofrequency ablation and mitral valvuloplasty, respectively. The effective doses during coronary angiography and coronary angioplasty, performed in room B, were found to be 4.6 and 10.2 mSv, respectively. A regional survey of the frequency of these cardiac procedures was performed. It was deduced that the annual collective effective dose from these cardiac procedures in the North of England, the former Northern Region, was 45.7 manSv. PMID- 9227232 TI - The interdependence of staff and patient doses in interventional radiology. AB - Staff doses arising from the use of X-rays are principally due to scattered radiation. This is related to the dose received by the patient expressed as the dose-area product (DAP). Doses to patients in interventional radiology are generally higher than for other fluoroscopically guided procedures. Doses to interventional radiologists are, therefore, amongst the highest associated with the use of diagnostic X-rays. The results of staff dose monitoring normalized to DAP should provide an indicator of those procedures which are associated with particularly high radiation exposures to staff, and should help to identify those radiologists whose practice may result in unnecessarily high doses to themselves. A study has been made of patient doses in two X-ray rooms used for interventional procedures associated with vascular and liver diseases. Doses to radiologists in these rooms were normalized to DAP. It was found that the average doses to the body, neck and hands were 0.05, 0.89 and 2.45 microSv/(Gy cm2), respectively for those radiologists with no significant involvement in hepatobiliary procedures. Higher doses were found for one radiologist whose workload included biliary drainage. The whole body dose was 0.17 microSv/(Gy cm2) or 5.8 mSv per year. It was shown that the doses to the neck and hands for the biliary drainage work was 6.59 and 29.0 microSv/(Gy cm2), respectively. This study has demonstrated the value of DAP as a measure of radiologist workload in respect of its significance in terms of staff dose. PMID- 9227233 TI - The effect of cortical endplates on ultrasound velocity through the calcaneus: an in vitro study. AB - Ultrasound velocity has been reported as a good predictor of bone strength measured in vitro using standard mechanical testing techniques. Such mechanical investigation of bone strength cannot be carried out in vivo, because of the invasive nature of the testing. Therefore to be able to extrapolate the in vitro findings to the clinical situation, the effect of cortex on ultrasound transmission velocity through the calcaneus is required. This was investigated in vitro by measuring ultrasound velocity through samples of different modification using a CUBAResearch ultrasound machine. The different sample modifications were: "whole" (soft tissue removed), "core" (cylindrical sample), "can" (cancellous sample without the cortex) and "def" (defatted cancellous sample). Ultrasound transmission velocity for the various sample modification were highly correlated with each other (r = 0.80-0.97). Coring resulted in a 0.77% increase in the mean velocity. Substituting bone marrow (defatting) with water at room temperature had no measurable effect on the ultrasound velocity. The velocity in the whole samples and the cancellous samples were statistically different with the cortex introducing only a 2% increase in the ultrasound velocity. Therefore the in vivo ultrasound velocity measured at the calcaneus is determined mainly by the cancellous bone component which is more sensitive to osteoporotic changes. Hence the reported ability of ultrasound velocity in vitro to predict bone strength could be expected in vivo. PMID- 9227234 TI - Radiotherapy in the treatment of solitary plasmacytoma. AB - Solitary plasmacytoma of bone (SPB) and extramedullary plasmacytoma (EMP) are rare. High local control rates are reported with radiotherapy, although the optimal dose and extent of radiotherapy portals remains controversial. Between 1983 and 1993, 30 patients with solitary plasmacytoma were seen at the Regional Cancer Centre, Trivandrum, India. 23 patients had SPB and seven EMP. The mean age was 52 years and the male to female ratio 3.2:1. Diagnosis of SPB was confirmed by biopsy in 16 patients and tumour excision in seven. 20 patients received megavoltage radiotherapy to the bone lesion with limited margins, and one received chemotherapy. Two patients who underwent complete tumour excision received no further treatment. All seven patients with EMP received megavoltage radiotherapy, four following biopsy and three after tumour excision. Local control was achieved in all patients with SPB. Nine progressed to multiple myeloma and one developed a solitary plasmacytoma in another bone. Six patients with EMP achieved local control. Three later progressed to multiple myeloma and one had local relapse. Median time to relapse was 28 months in SPB and 30 months in EMP. 5-year overall survival rates were 82% and 57% for patients with SPB and EMP, respectively. The corresponding progression free survival rates were 55% and 50%, respectively. Age, sex, site of tumour, serum M protein and haemoglobin levels did not significantly influence progression free survival. The extent of surgery, radiotherapy dose or time to relapse were not significant prognostic factors. Radiotherapy appears to be an effective modality of treatment of solitary plasmacytoma. No dose-response relationship is observed, and high local control rates are achieved with limited portals. Progression to multiple myeloma is the commonest pattern of failure, although no prognostic factors for progression are identified. The role of chemotherapy in preventing disease progression needs further evaluation. PMID- 9227235 TI - Predicting late rectal complications following prostate conformal radiotherapy using biologically effective doses and normalized dose-surface histograms. AB - A model to predict the late normal tissue complication probability (NTCP) of the rectum following conformal therapy is described. The model evaluates the biological consequence of inhomogeneities in the physical dose by computing dose histograms of the biologically effective dose to the surface of the rectum for a given fractionation scheme. A method of normalizing the surface area of the rectum is employed so that the predicted NTCP is independent of the differing cross-sectional size of sections of the rectum, ensuring the NTCP is dependent only on the dose delivered to sensitive rectal tissues. The model has been used to assess severe late rectal complications and the milder RTOG grades 2 and 3 reactions. This model was found to predict severe toxicity levels of 1.7 +/- 0.6% for an accelerated treatment of 50 Gy in 16 fractions commonly employed at this centre. This result lies between the severe toxicities predicted for 60 and 62 Gy delivered in 2 Gy fractions. The model predicts that the average NTCP for severe late effects for nine prostate patients becomes greater than 5% with a fractionation scheme of 70 Gy in 35 fractions, for the four fields treatment. The effects of not treating all fields at each therapy session on rectal toxicity were also investigated. Biologically effective dose-surface histograms show that the dose to the lower surface of the rectum is increased by not treating all fields at each therapy session, but the predicted differences in rectal NTCP are negligible. PMID- 9227236 TI - MRI of multiple biliary hamartomas. AB - Multiple biliary hamartomas (MBH) are uncommon benign malformations of the bile ducts. A case of MBH simulating liver metastases is reported, the diagnosis having been ascertained by US-guided biopsy. US and CT showed multiple hepatic nodules scattering in both lobes of the liver. Angiography showed small hypovascular lesions as well as hypervascular nodules. Only two cases with MRI studies have previously been reported. In this case, the finding was of hepatic cysts containing intermediate signal mural nodules. Demonstrating hepatic cysts with mural nodules on imaging will considerably increase the accuracy of diagnosis. PMID- 9227237 TI - Adenoid cystic carcinoma of the external auditory canal: correlation between histological features and MRI appearances. AB - A rare case of adenoid cystic carcinoma of the external auditory canal with magnetic resonance imaging appearances is reported. Both T1 weighted and T2 weighted MR images showed the tumour as a hypointense mass, although there was marked contrast enhancement. Microscopic examination of the resected tumour showed a preponderance of solid tumour cell nests. According to previous reports, these pathological and radiological findings indicate a poor prognosis. PMID- 9227238 TI - Obliteration of maxillary and sphenoid sinuses in Gaucher's disease. AB - Paranasal sinus obliteration is described in a patient with adult type Gaucher's disease. Plain radiographs and computed tomography showed obliteration of paranasal sinuses due to medullary expansion of surrounding bone. The mandible and maxilla are rarely affected in Gaucher's disease and obliteration of paranasal sinuses due to bony expansion has not previously been reported. PMID- 9227239 TI - Uncommon features of abdominal aortoiliac disease. AB - This pictorial review illustrates the cross-sectional imaging of several less common manifestations of aortoiliac disease. Despite their varying clinical features, the imaging appearances with ultrasound, CT and MRI will allow a correct assessment in most cases. PMID- 9227240 TI - The sheep in wolf's clothing. PMID- 9227241 TI - Shoulder dislocation as a complication of CT scanning. PMID- 9227242 TI - Imaging of anorectal function. PMID- 9227243 TI - Pontine haemorrhage following iohexol lumbar myelography. PMID- 9227244 TI - Standing conference on health and safety in the nuclear age. Report on a conference organized by Directorate General XI of the European Commission, held in Luxembourg, 26-27 November 1996. PMID- 9227245 TI - Fourth radiotherapy standards users meeting. Report on a meeting organized by the National Physical Laboratory, held on 9 July 1996. PMID- 9227246 TI - MRI of inner ear and facial nerve pathology using 3D MP-RAGE and 3D CISS sequences. AB - The aim of this study was to evaluate 3D CISS, unenhanced 3D MP-RAGE and contrast enhanced 3D MP-RAGE for the diagnosis of neoplastic, vascular and inflammatory lesions of the cerebellopontine angle, the inner auditory canal, the labyrinth and the facial nerve 42 MR examinations were performed on a total of 38 patients (25 males, 13 females; aged 1-77 years, mean age 43 +/- 20 years) using a 1.5 T MR unit. A T2* weighted 3D CISS sequence (TR 14.65 ms, TE 21 ms, flip angle 65 degrees, voxel size 0.7 x 0.7 x 0.7 mm3) and a T1 weighted 3D MP-RAGE sequence (TR 12.5 ms, TE 5 ms, T1 300 ms, flip angle 15 degrees, voxel size 1.0 x 0.9 x 0.9 mm3) with and without contrast medium (gadolinium-DTPA, 0.1 mmol kg-1 body weight) were used. Results of contrast enhanced 3D MP-RAGE-pathological enhancement was found in the following lesions: schwannomas of the cerebellopontine angle (CPA) and the internal auditory canal (IAC), 4; schwannomas of the IAC, 7 and labyrinthine tumours, 3; posterior fossa lymphoma, 1; meatal meningioma, 1; acute labyrinthitis, 15 and neuritis of the seventh cranial nerve, 10. Results of 3D CISS-filling defects were found with the following lesions: schwannomas of the CPA, the IAC or labyrinth, 14; lymphoma, 1; meningioma, 1; labyrinthine fibrosis, 13 and scar in the IAC, 4. These results suggest that unenhanced and contrast enhanced 3D MP-RAGE and 3D CISS are complementary MR imaging modalities. T1 weighted 3D MP-RAGE is preferred to T1 weighted 2D (turbo) spin echo sequences because of the multiplanar reconstruction possibilities of 3D sequences, which are very useful in the case of the inner ear and facial nerve. PMID- 9227247 TI - Early detection of locally recurrent rectal cancer by endosonography. AB - After curative surgery for rectal cancer, the goal of an aggressive surveillance programme is the detection of local recurrence (LR) at an early and potentially curable stage 62 patients (mean age 66.2 years) operated on for rectal cancer were prospectively enrolled in a follow-up study including endorectal ultrasound (EUS), serial CEA levels, digital examination, colonoscopy and pelvic CT. A total of 192 sonographic scans were performed, with a mean of three (range 2-7) for each patient. LR occurred in 11 patients; in all cases this was suggested by EUS. In two patients (18%) other techniques had failed to detect recurrent disease, which was identified solely by EUS. These two were treated radically and the remainder received radiotherapy or other palliative management. Five patients are alive at, on average, 18 months after LR (range 4-26 months). These include both cases treated with salvage surgery and who remain disease free. EUS is a valuable tool in the detection of locally recurrent rectal cancer. PMID- 9227248 TI - The incidence of coronary artery calcification on standard thoracic CT scans. AB - Coronary artery calcification indicates the presence of atheromatous disease and can represent an area of severe stenosis. The absence of calcification does not correlate with the absence of coronary artery disease, but an incidental finding of calcification has important prognostic implications. The best method available at present is electron beam computed tomography (EBCT) but these are often dedicated units in specialist centres and currently this modality is not widely available in the UK. This is the first study to establish the age/sex frequency with which incidental coronary artery calcification is detected using standard CT scanning in a large population. The methods used are simple, reproducible and based on standard thoracic protocols. The presence of calcification is easily detected using readily available equipment. The reported incidence of calcification using EBCT, the most sensitive method available, was compared with standard CT detection rates. While standard CT underestimates the presence of calcification, the age/sex trends are very similar. Standard CT probably detects the most dense lesions, missing areas of mild calcification, recent studies having shown that these lesions are most likely to represent areas of stenosis. This study shows that, with very little change in current practice and on readily available equipment, it is possible to obtain important diagnostic information suggesting the presence of severe coronary artery disease. PMID- 9227249 TI - Measurement of femoral antetorsion and tibial torsion by magnetic resonance imaging. AB - Several methods for measuring femoral and tibial torsion by CT have been described since 1980. Alternative methods of measurement to CT are needed to avoid irradiating young patients facing derotation osteotomy. In this study, MRI was used to quantify femoral and tibial torsion in normal adult volunteers, firstly with transverse sections analogous to those of CT in order to establish reference values; and secondly, with sections along the axis of the neck of femur to assess the influence of the orientation of the slices on the measured values. The images were acquired using fast T1 weighted gradient echo sequences in 0.2 and 1 T magnets. Average femoral antetorsion (CT-analogous method) was 10.4 degrees (average side difference = 4.6 degrees); average tibial torsion was 41.7 degrees (average side difference = 6.1 degrees). A steeply inclined slice along the axis of the femoral neck gave a mean measurement of the angle of antetorsion that was significantly higher than the mean obtained from either slightly inclined or transverse sections (16.7 degrees vs 12.1 degrees and 11.2 degrees, respectively, p < 0.001). In conclusion, MRI provides an alternative to CT in the measurement of femoral and tibial torsion. MRI enables one to orientate the slice along the axis of the femoral neck, thus obtaining a single cross-section of the entire neck. However, the normal range of measurements will vary according to the plane of image. PMID- 9227251 TI - Assessing the degree of osteoporosis in the axial skeleton using the dependence of the fractal dimension on the grey level threshold. AB - Combining the measurement of bone mineral density (BMD) and the classification of the trabecular structure in cancellous bone improves the estimation of the degree of osteoporosis. A fractal method for the automatic quantitative classification of the trabecular structure in midvertebral slices of lumbar vertebrae is introduced. This method is based on the computation of the fractal dimension (box counting method) for varying binarization thresholds. Radiographic images from 30 lumbar vertebrae and CT images from an additional 16 lumbar vertebrae were analysed by calculating the dimension D in dependency of the threshold value T. The function D(T) was normalized by the average image grey value, eliminating the bone mineral density from the computations. The results show that the images of the lumbar vertebrae have fractal properties, and the function D(T) has a typical behaviour that allows the discrimination of the degree of osteoporosis. With two parameters extracted from the function D(T) the correlation coefficients with BMD were both -79% for the radiographic images, and -93% and -91% for the CT data, respectively. PMID- 9227250 TI - Male and female normative data for ultrasound measurement of the calcaneus within the UK adult population. AB - The establishment of accurate normative data for specific populations is required before ultrasound bone densitometry may be considered for routine clinical application. A single general practice was utilized, representing both rural and urban populations, to recruit 169 male and 210 female subjects in the age range 20-80 years. Measurements of broadband ultrasound attenuation (BUA) and velocity (VOS) of the left calcaneus were undertaken using the McCue CUBAClinical II system. Two descriptive statistical models were used, mean and 95th percentile, and regression analysis. The female data for both BUA and VOS illustrated peak values in the 30-39 years age group, falling steadily thereafter. For male BUA and VOS data there was less evidence of an age related dependence. Upon analysis of previous normative studies and current data it is evident that there is considerable variability. In addition to different commercial systems being utilized, there is a lack of consistency in study design and method of subject recruitment. The need for accurate calibration and cross-calibration methods for commercial ultrasound bone densitometers is therefore of paramount importance to enable clinical interpretation of ultrasound measurements to be performed. PMID- 9227252 TI - Serum PICP as a bone formation marker in 89Sr and external beam radiotherapy of prostatic bony metastases. AB - The clinical management of skeletal metastatic disease is problematic because of the difficulty in treating and accurately monitoring therapy impact and disease progression. This investigation measured serum procollagen type I C-terminal peptide (PICP) concentrations as a semi-quantitative index of bone turnover in patients with metastatic prostatic adenocarcinoma before and following palliative 89Sr chloride therapy. 10 patients with early stage (stage A2, B1 and B2) biopsy confirmed prostatic adenocarcinoma were investigated (n = 10). Two groups of 10 patients each (n = 10 per group) with advanced (stage D) metastatic prostatic adenocarcinoma who had previously undergone hormonal manipulation were also investigated. One group of patients with scintigraphically documented metastatic bone disease received additional irradiation for new symptomatic bone metastases, whereas the other group received 89Sr chloride therapy. A radioimmunoassay for PICP was used to measure serum concentrations of patients in each of these groups as well as positive and negative controls. The concentration of serum PICP rose from 649 +/- 279 before treatment with external beam radiotherapy to 927 +/- 157 ng ml-1 4 months after therapy (p < 0.05). However, the results demonstrated a four-fold decrease (p < 0.001) in serum PICP in clinical responders to 89Sr chloride therapy versus baseline 4 months after the completion of treatment. The clinical non-responders demonstrated no significant change in PICP concentrations during that interval. This may be due to an increase in untreated bony metastases in the non-89Sr treated group. Although a relatively small representative group of patients was studied, these data demonstrate that serum PICP concentration correlates with clinical response to 89Sr chloride therapy. This objective laboratory technique may be useful for monitoring and predicting the need for 89Sr chloride therapy and optimizing palliative care. It may also be extremely useful in predicting a therapeutic response to such intervention. PMID- 9227253 TI - Modification of bone marrow radiosensensitivity by medicinal plant extracts. AB - Withaferin A (WA), a steroidal lactone, and Plumbagin (Pl), a naphthoquinone, from the roots of Withania somnifera and Plumbago rosea, respectively, have been shown to possess growth inhibitory and radiosensitizing effects on experimental mouse tumours. An aqueous extract of the leaves of Ocimum sanctum (OE) was found to protect mice against radiation lethality. Therefore, the radiomodifying effects of the above plant products on the bone marrow of the adult Swiss mouse was studied. Single doses of WA (30 mg kg-1) or Pl (5 mg kg-1) were injected intraperitoneally (ip) and OE (10 mg kg-1) was injected ip once daily for five consecutive days. Administration of extracts was followed by 2 Gy whole body gamma irradiation. Bone marrow stem cell survival was studied by an exogenous spleen colony unit (CFU-S) assay. The effects of WA and Pl were compared with that of cyclophosphamide (CP) and radioprotection by OE was compared with that of WR-2721 (WR). Radiation reduced the CFU-S to less than 50% of normal. WA, CP and Pl significantly enhanced this effect and reduced the CFU-S to almost the same extent (to < 20% of normal), although individually WA and Pl were less cytotoxic than CP. These results indicate that radiosensitization by WA and Pl is not tumour specific. OE significantly increased CFU-S compared with radiotherapy (RT) alone. OE+RT gave a higher stem cell survival (p < 0.05) than that produced by WR+RT. While WR alone had a toxic effect, OE treatment showed no such effect, suggesting that the latter may have an advantage over WR in clinical application. PMID- 9227254 TI - Human biodistribution of an ultrasound contrast agent (Quantison) by radiolabelling and gamma scintigraphy. AB - The biodistribution and kinetics of an air filled human serum albumin microcapsule formulation (Quantison) intended for use as an intravenous ultrasound contrast agent have been examined. 12 healthy subjects were administered with approximately 50 million microcapsules per kilogram body weight, radiolabelled with 50 MBq 123I. Imaging was performed over a period of 58 h using a large field-of-view gamma camera and the amount of labelled material present in the blood, urine and faeces measured. Imaging demonstrated that the liver was the organ with the highest uptake, with a mean uptake of 41.8% (SD 10.4%) of the administered dose 1 h following administration. The maximum uptake of the agent in the lungs was low, mean 4.0% (SD 3.4%). A small amount of uptake was visible in the bone marrow; however, this was not quantifiable. There was also evidence of minimal myocardial activity within 5 min of administration. No adverse events were observed and there were no changes in any of the individual post-study indices. The present study demonstrates the safety of Quantison. Gamma scintigraphy played a useful role in confirming the biodistribution of the agent with little lung uptake, high liver uptake and evidence of myocardial uptake. PMID- 9227255 TI - Quality control programme in mammography: second level quality controls. AB - Mammography is the most reliable method by which to detect lesions in the breast. Since contrast between normal and pathological areas in the breast is extremely low, mammographic image quality should reach high standards without exceeding acceptable exposure levels for the breast. A quality control programme in mammography has been implemented. This programme is subdivided into two levels. The first consists of simple daily checks of image quality and film processing, while the second deals with more complex checks of mammographic unit, screen-film system, darkroom, illuminators, viewing conditions and reference dose determination. The values of all the parameters undergoing measurement are compared with the limiting values given by National and International Protocols. This paper describes the second level controls carried out every 6 months by the medical physicist. The parameters described are only those which have been studied and analysed in detail since the quality control programme in mammography was implemented. Such parameters (kilovoltage, focal spot dimension, half value layer, tube output, automatic exposure control system, screen-film characteristic curve and mean glandular dose) were measured during the period 1991-1995 and the results summarised. The values obtained prove the constant correct functioning of the equipment. PMID- 9227256 TI - Assessment of lifetime gained as a result of mammographic breast cancer screening using a computer model. AB - A computer model for the simulation of breast cancer screening (MBS) is used to calculate the results of screening in terms of lifetime. To optimize breast cancer screening protocols, risk (lifetime lost due to radiation-induced tumours) versus benefit (lifetime gained due to early detection of breast cancer) analyses are performed for a simulated stable Swedish female population. The present study focuses on the results of different screening strategies employing single view mammography, including starting and finishing ages of screening, time interval between successive screening sessions as well as the influence of high detection screening and differences between different populations, based on lifetime lost or gained. To test the stability of the recommendations with respect to possible changes in the variables used in MBS, calculations are also performed for high risk factors for breast tumour induction using both the additive and multiplicative risk models, fast growing breast tumours, late incidence of breast tumours and age dependent survival. The results of the simulations expressed in terms of lifetime gained suggest that a theoretical benefit can be obtained employing starting and finishing ages of 35 and 75 years, respectively. In terms of number of fatal breast tumours, the favourable screening period is 40-80 years. It is concluded that the recommendations are stable for changes in the input variables of MBS. The benefits of higher detection screening are more marked for younger than for older women. A high screening frequency results in more lifetime gained, especially at relatively young ages, whereas for older ages the effect is only marginal. PMID- 9227257 TI - Evaluation of a new ultraviolet-emitting rare-earth film-screen combination. AB - The performance of a 400 speed class DuPont Ultra Vision Rapid (UVR) film-screen combination has been evaluated and compared with that of DuPont Quanta Fast Detail screens with Cronex 10L film (QFD-200 speed class). The speed was calculated from the constructed characteristic curves (H&D) at different energies. Image quality was derived objectively using the Leeds test object TOR (CDR). An anthropomorphic phantom was then employed to determine image quality subjectively by means of radiologist appraisal. Lumbar spine and chest radiography of patients were performed to evaluate the potential for dose reduction in clinical conditions by measuring skin entrance doses with thermoluminescent dosimeters (TLDs). UVR provided better resolution (9 lp mm-1 as opposed to 8 lp mm-1) and contrast response than QFD. UVR has accommodated a wider exposure latitude than might be expected with conventional 400 speed class film-screen combinations. Use of UVR resulted in better image quality than use of QFD over a wide range of exposure factors. The use of UVR can result in a dose reduction of 50% with no loss in image quality. PMID- 9227258 TI - Knee tumours--duration and nature of symptoms prior to investigation. AB - The incidence of most musculoskeletal neoplasms is highest around the knee. Royal College of Radiologists Guidelines for diagnostic imaging state that plain radiographs are not routinely indicated for knee pain without restriction of movement or symptoms of locking. The notes of 19 patients with knee tumours presenting over a 5 year period were analysed. Irrespective of age or the grade of malignancy the majority of patients had symptoms for around 6 months prior to the initial radiograph. Only four patients had symptoms which would have merited radiological investigation under the present Guidelines. Even in these patients the symptoms prior to either pathological fracture or locking would not have come under the Guidelines. There is a case for regarding persistent, unilateral knee pain for longer than 6 weeks as an indication for imaging. PMID- 9227259 TI - Monitor unit calculation in 6 MV irregularly shaped beams--accuracy in clinical practice. AB - The results of an investigation of the accuracy of monitor unit (MU) calculation in clinical shaped beams are presented. Measured doses at the reference depth on the beam central axis (isocentre) or on a beam axis representative of the irradiated area (when the isocentre lies under a block or near the edges of the block's shadow) were compared with the expected doses when calculating MUs, by applying different methods normally used in clinical practice. Empirical (areas weighted, Wrede) and scatter summation (Clarkson) methods as well as a pencil beam based algorithm were applied. 40 irregular fields (6 MV X-rays, CLinac, Varian 6/100), divided into six categories, were considered. Dose measurements were performed with a NE2571 ionization chamber in an acrylic 30 x 30 x 30 cm3 phantom. The depths in acrylic were converted into water-equivalent depths through a correction factor derived from TMR measurements. The method of dose measurements in acrylic was found to be sufficiently accurate for the purpose of this study by comparing expected and measured doses in open square and rectangular fields (mean deviation +0.2%, SD = 0.5%). Results show that all the considered methods are sufficiently reliable in calculating MUs in clinical situations. Mean deviations between measured and expected dose values are around 0 for all the methods; standard deviations range from 1% for the Wrede method to 0.75% for the pencil-beam method. The differences between expected and measured doses were within 1% for about 3/4 of the fields when calculating MUs with all the considered methods. Maximum deviations range from 1.6% (pencil-beam) to 3% (Wrede). Slight differences among the methods of MU calculation were revealed within the different categories of blocked fields analysed. The surprising agreement between measured and expected dose values obtained by using empirical methods (area weighted and Wrede) is probably due to the fact that the reference points were positioned in a "central" region of the unblocked areas. PMID- 9227260 TI - Sagittal vertebral body fractures: magnetic resonance imaging features. AB - Isolated sagittal vertebral body fractures are rare and the plain radiographic diagnosis may be difficult. We report two cases, confirmed by CT, and describe the subtle MRI features which comprised increased signal intensity only on the midline sagittal T2 weighted images. In one case, the information from MRI significantly altered the patient's management by leading to a change from surgical to conservative treatment. PMID- 9227261 TI - Hydatid cyst of the liver communicating with the left colon. AB - Rupture and secondary infection are common complications of hydatid cyst in the liver. Ultrasound and CT findings are reported in a case of hydatid cyst which has ruptured directly into the left colon. Rupture of hydatid cyst into a hollow viscus is extremely rare. CT demonstrated partial drainage of the cyst contents with the creation of an air-fluid level. PMID- 9227262 TI - Transvaginal leak of peritoneal dialysate demonstrated by CT peritoneography. AB - CT peritoneography is the investigation of choice for suspected leaks or hernias in patients on continuous ambulatory peritoneal dialysis (CAPD). We report a case of transvaginal leak of peritoneal dialysate in a young woman, confirmed by CT peritoneography. CAPD has been successfully recommenced following the repair of a left vaginal vault erosion. PMID- 9227263 TI - Magnetic resonance imaging of an ovarian mucinous cystadenoma immediately before and after rupture. AB - The MRI findings are described in a case of rupture of an ovarian mucinous cystadenoma which occurred during the course of an MR scan. While the rupture was presumed to be spontaneous, possible precipitating factors are discussed. PMID- 9227264 TI - Segmental arterial mediolysis studied by repeated angiography. AB - Segmental arterial mediolysis (SAM) is a rare disease of unknown aetiology. We report the fourteenth case of SAM, but the first to demonstrate serial changes on arteriography. A 65-year-old woman with abdominal pain underwent laparotomy with resection of an abnormally beaded and narrowed segment of the right branch of the middle colic artery. Characteristic pathological findings of lysis of the arterial media with dissecting haematomas were present. Other than some post prandial pain, the patient's post-operative course was uneventful. Serial arteriography showed various abnormalities in the trunk and branches of the superior mesenteric artery. Changes in the vessels consisted of three phases, i.e. dilatation, beading with narrowing and restoration of the smooth wall, with various modifications such as aneurysmal enlargement and occlusion. PMID- 9227265 TI - Multiple intraabdominal aneurysms--getting to the heart of the matter. PMID- 9227266 TI - Accidental irradiation of electronics used for TBI monitoring. PMID- 9227267 TI - Tear hepatocyte growth factor (HGF) availability increases markedly after excimer laser surface ablation. AB - Tear cytokines and growth factors are likely to modulate the wound healing process following corneal epithelial injury. Hepatocyte growth factor (HGF) is a paracrine mediator of epithelial proliferation, motility, and differentiation that is produced by keratocytes and the lacrimal gland. Tear samples were collected preoperatively and one, two, and seven days postoperatively in eyes undergoing excimer laser surface ablation [photorefractive keratoplasty (PRK) or phototherapeutic keratoplasty (PTK)]. Tear HGF concentration was measured with a sensitive ELISA assay. Tear HGF production was calculated using the tear flow rate in the collection capillary and HGF concentration. Although the instantaneous concentration of HGF in tears decreased significantly in the days following PRK, a large increase in tear flow resulted in a marked increase in HGF bioavailability. The heparin-binding characteristics of HGF would result in increased binding to glycosaminoglycans and other heparin-like matrix components and, therefore, increased growth factor availability to the cognate recptor. This is the first report documenting changes in tear film HGF production. HGF may have an important function in maintenance and wound healing of the ocular surface epithelium since HGF is present in the normal tear film and the HGF secretion rate increases markedly in parallel with aqueous tear production following corneal surgical injury. PMID- 9227268 TI - Intravitreal sustained release of VEGF causes retinal neovascularization in rabbits and breakdown of the blood-retinal barrier in rabbits and primates. AB - Vascular endothelial growth factor (VEGF) has been identified as a possible mediator of retinal neovascularisation (NV), but it is not certain if VEGF alone is sufficient to cause retinal NV. We sought to investigate this issue by implanting ethylene-vinyl acetate copolymer pellets that slowly release VEGF into the vitreous cavity of rabbits and primates. Eyes were examined by indirect ophthalmoscopy, fundus photography, and fluorescein angiography and then animals were killed at various time points and immunocytochemical and ultrastructural evaluations were carried out. Seven days after implantation of a pellet containing 30 micrograms of human recombinant VEGF into the vitreous cavity of rabbits, retinal blood vessels became dilated and tortuous, and between days 14 and 21, retinal NV was noted in all eyes. Fluorescein angiography showed profuse leakage of dye from the anomalous vessels. Immunohistochemical staining for proliferating cell nuclear antigen (PCNA) showed positively staining nuclei in many of the endothelial cells of new blood vessels on the surface of the retina. Six eyes implanted with control pellets containing vehicle and two eyes implanted with pellets containing 30 micrograms of human serum albumin alone showed no retinal vascular abnormalities. Implantation of pellets containing 100 micrograms of VEGF into the vitreous cavity of primates resulted in iris NV and retinal vascular dilation and tortuosity very much like that seen in humans with ischemic retinopathies. Immunohistochemical staining for serum albumin showed widespread severe breakdown of the blood-retinal barrier (BRB). Histology showed dilated thin-walled retinal vessels, but unequivocal retinal NV could not be identified and staining for PCNA was negative. These findings indicate that sustained intravitreal release of VEGF causes widespread retinal vascular dilation and breakdown of the BRB. Retinal NV seems to require persistent high levels of VEGF at the retinal surface and can be achieved in rabbits providing a potentially useful model of retinal NV, but is difficult to achieve in primates. The extensive VEGF-induced disruption of the blood-retinal barrier suggests that VEGF antagonists may provide a new therapy for patients with ischemic retinopathies and macular edema. PMID- 9227270 TI - Activation of calpain in lens: a review and proposed mechanism. AB - The purpose of these experiments was to develop a hypothesis to explain activation of m-calpain in cataractogenesis observed in rodents. The in vitro model used to study m-calpain activation was to correlate breakdown of the 'reporter' protein alpha-crystallin with the appearance of activated m-calpain using protein sequencing and casein zymography. Incubation of alpha-crystallins with m-calpain and Ca2+ caused proteolysis of alpha-crystallins and accumulation of new polypeptides. E64 and calpain inhibitor I each inhibited proteolysis of alpha-crystallins. The N-terminus of the 80 kDa subunit of m-calpain was blocked at time 0 (pro calpain). After incubation with Ca2+, the remaining 80 kDa subunit of m-calpain gave a N-terminal sequence of KDREAAEGLG, indicating loss of nine amino acid from the N-terminus (autolysed calpain). The new 43 kDa m-calpain fragment also gave a N-terminal sequence of KDREAAEGLG, indicating the same loss of the first nine amino acids on the N-terminus as well as a major loss of the C terminal half of the subunit (degraded calpain). In contrast, the N-terminus of the 80 kDa subunit of m-calpain remained blocked when E64 was present (unautolysed form). Moreover, the Ca2+ concentration required for proteolysis decreased when calpain was pre-incubated with Ca2+, although proteolysis of alpha crystallin required a higher Ca2+ concentration than proteolysis of casein. These data suggested that the sequence of events for m-calpain activation were unautolysed, autolysed and finally degraded calpain. Unautolysed and/or autolysed calpains may be proteolytically active against alpha-crystallin. PMID- 9227269 TI - Human papillomavirus-16 E6/E7 transfected retinal cell line expresses the Muller cell phenotype. AB - The introduction of viral transforming genes into mammalian cells has been used in establishing cultures of unlimited lifespan. Although Muller cells, the predominant glial cells in the mammalian retina, have been isolated using a variety of techniques, most of these cultures have limited capacity for cell division and are often contaminated by other cell types especially astrocytes, endothelial cells and microglial cells. We have established pure cultures of retinal cells which express Muller cell characteristics and exhibit unlimited growth in vitro. We now report the techniques involved in the propagation and characterization of these cultures. Mixed retinal cultures isolated from dystrophic rat retinas were infected with defective retroviruses coding for human papillomavirus (HPV) type 16 E6 and E7 proteins. The disabled viral constructs also contained the neomycin gene allowing selection of the cultures using Geneticin, a neomycin analogue. Pure cultures were then obtained from Geneticin selected populations by limiting end-dilution techniques. The expression of the HPV-16 E6/E7 genes in the transfected cell line was established using an HPV-16 E6/E7 PCR product to probe Northern blots. Cloned cells were found to be highly reactive for Muller cell markers including S-100, carbonic anhydrase-C, cellular retinaldehyde binding protein, and glial fibrillary acidic protein but not for glutamine synthetase. Ultrastructural studies showed stacks of cells with long elaborate processes, short microvilli, coated pits, cytoplasmic filaments, abundant perinuclear rough endoplasmic reticulum, and smooth endoplasmic reticulum extending to the cell processes. Growth patterns of late passage cells (> 50 passages) showed a lag phase of 48 hr followed by exponential growth extending past visual confluence at day 5. Since the cultures have undergone more than 240 population doublings, they can be characterized as a continuous cell line with unlimited lifespan. The HPV-16 E6/E7 transfected Muller cell line may prove useful in studies requiring abundant and pure cultures of Muller cells. PMID- 9227271 TI - Dexamethasone treatment decreases hyaluronan-formation by primate trabecular meshwork cells in vitro. AB - The functional significance of Hyaluronan (HA) present in the cribriform layer of Schlemm's canal is not known. It may contribute to the actual outflow resistance but the relatively inert molecule might also be necessary to prevent adherence of larger molecules to the cribriform network. Thus HA might rather prevent an increase in outflow resistance. It is well known that treatment with Dexamethasone (DM) in a number of patients leads to an increase in intraocular pressure presumably due to an increase in outflow resistance. To clarify whether an imbalance in HA formation might be involved in these changes we have treated confluent cultures of human trabecular cells as well as control cell lines (ciliary muscle cells, scleral fibroblasts) with 500 nM DM for 24 hr or 12 days and have measured HA-synthesis using incorporation assays with 0.05 m D-[6-3H] Glucosamine hydrochloride. In all six cell lines investigated there was a significant decrease in HA-synthesis following short and long term treatment with DM when compared with the untreated controls. This reaction of trabecular cells to DM treatment is different from the DM effect reported on the synthesis of many other components of the extracellular matrix like fibronectin and elastin which increase after DM treatment. If the DM-effect seen in cell cultures plays a role in vivo decreased formation in HA could result in impaired function of the outflow pathways. PMID- 9227273 TI - Inheritance and strain distribution of a persistent hyaloid vascular system in mice. AB - The mode of inheritance of a persistent hyaloid vascular system was investigated in an inbred strain of Senescence-Accelerated Mouse P9 (SAMP9) by conducting crosses between SAMP9 and SAMR1, a strain which shows normal regression of the hyaloid vascular system. We also examined the distribution of this abnormality in 12 inbred SAM strains and in eight commonly used inbred strains of mice. Ophthalmoscopic examination of the eyes of 5-week-old mice, which have transparent lenses, revealed the persistence of a hyaloid vascular system in only one female F1 hybrid out of 66 offspring. The observed segregation ratio of affected to unaffected mice was 25:52 in males and 37:44 in females, following the reciprocal backcross progeny between SAMP9 mice and F1 hybrids. The results of the strain distribution study indicated that 8-97% of the mice among six strains of SAM exhibited the persistence of a hyaloid vascular system, whereas the other inbred strains did not exhibit this abnormality. These observations suggest that at least two major genes may contribute to the persistence of a hyaloid vascular system, and suggest that the SAM strains comprise a group of related inbred strains. PMID- 9227272 TI - Analysis of variance of microspheres blood flow measurements in rabbits. AB - As part of a larger study on the interpretation of angiographically derived hemodynamic parameters, blood flow in several ocular tissues was measured using the radioactively labelled microspheres technique. As an unexpected secondary results, it was found that the microspheres data gave quantitative information on hyperaemic effects in the eye. This is the subject of the present paper. The measurements were made in 13 anaesthetized pigmented rabbits. In each animal, three blood flow measurements were performed at three different ocular perfusion pressures (60-15 mmHg). The perfusion pressures of the experimental eye were varied by changing the intra-ocular pressure. The contra-lateral eye served as a control. Labelled microspheres were used as a non-recirculating blood flow indicator, enabling the estimation of regional blood flows, in this case for the iris, ciliary body, peripheral choroid and peripapillary choroid separately. Using analysis of variance with perfusion pressure as covariate and taking into account the blood flow of the control eye, hyperaemia could be quantified in the experimental eye. Apart from a difference amongst animals, hyperaemia depended on tissue type. The amount of hyperaemia proved to be more pronounced in the anterior part of the eye, iris and ciliary body, and to decrease towards the posterior pole. With regard to the causes of this hyperaemia one could speculate about the invasive handling (anterior eye needles) topical administration of tropicamide, in combination with the general anaesthesia. PMID- 9227274 TI - Effects of antioxidants on ischemic retinal dysfunction. AB - Transient occlusion of both common carotid arteries in normotensive rats leads to a reduction in the amplitude of the b-wave of the electroretinogram (ERG). The present study investigated whether the antioxidants vitamin E and lipoate attenuate the depression and enhance the recovery of the b-wave of the ERG in response to retinal ischemia. The ERG was recorded from halothane-anesthetized rats before. during and after transient (24 min) occlusion of both common carotid arteries. The substances were administered 20 minutes before or at the onset of ischemia. Both vitamin E (100 mg kg-1) and lipoate (100 mg kg-1) significantly reduced the depression of the b-wave during occlusion and accelerated recovery during reperfusion at either time point of application. The present results suggest that antioxidants provide protection against ischemic retinal dysfunction. PMID- 9227275 TI - Confocal microscopy of cataracts from animal model systems: relevance to human nuclear cataract. AB - A recent study demonstrated that cytosolic lipid membrane structures, independent of the plasma membrane, preferentially occurred in human cataractous lenses. Animal model systems of cataractogenesis (selenite treated rats: galactose fed rats; buthionine-sulfoxime treated mice; Emory mice) were screened for possible relevant structures using the lipid membrane probe DiI and confocal microscopy. Well delineated plasma membranes of lens fiber cells with independent cytosolic staining structures were only observed in the selenite model system. These cytosolic structures were not observed in aged matched control lenses or within the transparent cortical regions of selenite treated animals with intense nuclear opacification. These results suggested that the morphological changes in DiI staining structures seen in the nucleus of the human cataractous lens were best approximated by those seen in the selenite model system. PMID- 9227276 TI - Unscheduled DNA replication precedes apoptosis of photoreceptors expressing SV40 T antigen. AB - Expression of simian virus 40 T antigen (Tag) in the rod photoreceptors of transgenic mice leads to cell death that is completed by the end of the third week of postnatal development. To understand the mechanistic link between Tag expression and the death of the expressing photoreceptors, cell cycle activity was followed in a transgenic mouse family that expresses Tag directed by the mouse opsin promoter. Tag-expressing photoreceptors also expressed rhodopsin suggesting that these cells were differentiated. The presence of Tag in the photoreceptors induced the expression of both proliferating cell nuclear antigen (PCNA) and thymidine kinase (TK). The abnormally high levels of PCNA and TK continued until the complete disappearance of the cells expressing Tag. Photoreceptor cell death was also associated with continued DNA synthesis that ceased shortly after postnatal day 16. The specific loss of the rod photoreceptors that re-entered the cell cycle accounted entirely for the loss of photoreceptors from the outer nuclear layer. The antiproliferative nature of the mature retina is directly involved in the apoptotic death of photoreceptors expressing Tag. PMID- 9227277 TI - Hyaluronic acid in the normal and glaucomatous optic nerve. AB - Eighteen normal human eye-bank eyes (age: 18-81 years), five fetal eyes (16-24 weeks), 11 primary open-angle glaucoma (POAG) eyes (age: 76-89 years), and two Schnabel's cavernous optic atrophy eyes were examined using a biotinylated hyaluronan binding protein to study the changes in the distribution of hyaluronic acid (HA) in the fetal, adult and glaucomatous optic nerve head. The vitreous body served as a positive control. Sections treated with Streptomyces hyaluronidase were used to confirm specificity. Monoclonal antibodies to myelin basic protein (MBP) and glial fibrillary acidic protein (GFAP) were used as additional controls. In fetal optic nerve, HA was localized in blood vessels, peripapillary sclera and the pial septae in the retrolaminar nerve. No staining was associated with axons. Staining for MBP was negative. In adults, HA was found surrounding the myelin sheaths in the retrolaminar nerve; staining decreased with age. In contrast, HA staining in myelinated peripheral nerves (e.g. ciliaries) remained unchanged with age. HA also was localized to the adventitia of arteries and veins throughout the posterior segment. Compared to age-matched normal eyes, HA staining was virtually absent around myelin sheaths of the retrolaminar nerve in POAG eyes. Similar changes were not found in other HA positive structures. In Schnabel's cavernous optic atrophy. HA was present in increased amount in the atrophic area, but virtually absent in the remaining retrolaminar nerve. HA staining was invariably positive in vitreous, and Streptomyces hyaluronidase treated sections were negative. In adults, staining of MBP was associated with the myelin sheath in the retrolaminar nerve. In contrast to HA, staining of MBP was unchanged with age and in POAG. In Schnabel's atrophy, MBP staining disappeared only in the atrophic area. HA in the retrolaminar optic nerve appears to be associate with the space-filling matrix between myelin sheaths. HA is not present in the axon bundles prior to myelination of the optic nerve. HA in the retrolaminar optic nerve appears to decrease with age and is further reduced in POAG; however, corresponding changes are not found in MBP or in peripheral nerves. Perhaps, decreased amounts of HA is related to a higher susceptibility to elevated intraocular pressure or to optic nerve atrophy. In Schnabel's cavernous optic atrophy, HA is present in increased amount only in the atrophic area while MBP is markedly decreased, suggesting in situ production of HA in areas of optic nerve atrophy. PMID- 9227278 TI - Demonstration of discrete secreted and membrane-bound ocular mucins in the dog. AB - Our aims were to separate and characterize secreted canine ocular mucins, and to provide definitive evidence of membrane-bound mucins at the canine ocular surface. Mucus was collected by suction from the ocular surface of normal dogs and dispersed in guanidine hydrochloride and a cocktail of protease inhibitors. Caesium chloride density gradient centrifugation separated secreted mucins from membranes, which were collected from the top of the gradients. Membranes were extracted with octyl glucoside and screened using lectins and anti-mucin antibodies. Gradient fractions containing secreted mucins were constituted into pools on the basis of differential lectin and antibody staining. High molecular weight material from each pool was purified by gel filtration. This material, and the membrane extract, were reduced and alkylated. Vacuum blotting of separated materials after agarose gel electrophoresis was used to compare subunit structure. Density gradient profiles indicated three principal secreted glycoprotein peaks: one staining strongly with anti-mucin antibodies. Gel filtration demonstrated that each contained high molecular weight material. Vacuum blots demonstrated the presence of two secreted glycoproteins with differently sized subunits. On the basis of buoyant density, one of these may be lipid complexed. Membrane extracted material stained with anti-mucin antibodies, and vacuum blotting of this material provided evidence for two membrane-bound components. In conclusion, we have shown that normal canine ocular mucus contains two secreted mucins, each exhibiting different subunit structure; one of these mucins may undergo lipid complexation. Normal canine ocular mucus also contains two membrane-bound mucins: one of which is unique among membrane mucins in showing subunit structure. PMID- 9227279 TI - Disulfide bond formation of cysteine-37 and cysteine-66 of beta B2 crystallin during cataractogenesis of the human lens. AB - Beta B2 crystallin has been prepared from total protein of normal (transparent) and cataractous human lenses. After digestion with endoprotease lys-C, the crude digests were resolved on a C18 reverse phase column. The lys-C digests of beta B2 crystallin from cataractous lenses consistently showed the presence of a major peptide that was not present in the lys-C digests from normal lenses. Treatment of this peptide with dithiothreitol resulted in the production of two peptides, corresponding to beta B2 crystallin sequences 18-41 and 48-67, containing cysteine-37 and cysteine-66, respectively. The results demonstrated that intramolecular disulfide bonding of these two residues had occurred in vivo. Based upon previous knowledge of the three dimensional structure of beta B2 crystallin, formation of this disulfide bond suggests that significant denaturation of the beta B2 crystallin molecule has occurred during the process of human lens opacification. PMID- 9227280 TI - Low de novo glutathione synthesis from circulating sulfur amino acids in the lens epithelium. AB - Transport of circulating sulfur amino acids (SAA) into the lens epithelium and de novo glutathione (GSH) synthesis were studied in the perfused guinea-pig eye. Plasma-to-aqueous transfer of SAA was in their intact form (> or = 98%) and comparable with sucrose (an extracellular marker) within 30 min. The unidirectional transport rates (ml min-1 g-1) of 35S-labeled cystine, cystine and methionine into the epithelium were: 0.0057, 0.0003 and 0.0073 from plasma, and 1.41, 0.005 and 1.69 from aqueous, respectively. The unidirectional epithelial uptake was limited to 1 min for all three [35S]SAA, and the isotopic steady-state ratio was achieved between 1 and 30 min. Cortical uptake was time-dependent and progressive between 1 and 30 min, but undetectable within 1 min. The high performance liquid chromatography (HPLC) analysis of the epithelium revealed that following 1 min of unidirectional [35S]cysteine transport, 3% of the label was incorporated into GSH and > or = 95% was as cysteine. An average incorporation of [35S]cysteine into GSH within the 30 min period was 0.83% min-1 and 1%/min for the epithelium and cortex, respectively. Infusions of [35S]cystine and methionine failed to demonstrate incorporation into GSH. Maximal rates of de novo GSH epithelial synthesis were approximately 3 and 12 pmol g-1 from plasma and aqueous cysteine, respectively. A t1/2 of 5480 hr was estimated if epithelial GSH had to be replaced exclusively by synthesis from aqueous cysteine. Given the limited aqueous and epithelial cysteine pools, and low (from cysteine) or undetectable (from cystine and methionine) incorporation of the label into GSH, we conclude that de novo GSH synthesis from circulating and aqueous SAA can be only a minor source of the millimolar concentration of GSH in the epithelium. PMID- 9227281 TI - Synergistic rise in Ca2+ produced by somatostatin and acetylcholine in ciliary body epithelial cells. AB - The purpose of these experiments was to demonstrate the presence of somatostatin receptors on the nonpigmented epithelial cells of the rabbit ciliary body and their link with intracellular Ca2+ homeostasis. Freshly excised rabbit ciliary processes and nonpigmented cell layer, explants were loaded with the fluorescent dye fura-2, and free-Ca2+ concentration ([Ca2+]i) in the nonpigmented cells was measured with fluorescence ratio imaging. The cells were continuously perfused, and drugs were added to the perfusate. Somatostatin-14 (SS14, 0.1-1.0 microM) or acetylcholine (ACh, 10 microM) applied alone produced small increases in [Ca2+]i. However, SS14 (0.1 microM) in combination with ACh (10 microM) induced a massive increase in [Ca2+]i (25.7 +/- 3.3 times the baseline level, n = 28). The dose response curve for SS14 (in the presence of 10 microM ACh) was sigmoidal with an EC50 of 3.9 nM and Hill coefficient of 2.5, indicating the requirement for multiple SS receptor activation. Somatostatin-28 could mimic the effect of SS14, although a much higher concentration was required. Shifting the SS14 dose response curve to the right by about two-orders of magnitude resulted in a fit to the SS28 data. The response to ACh + SS14 could not be blocked by the alpha 2 adrenergic blocker yohimbine (Yoh, 10 microM) or the A1-specific adenosinergic antagonist 8-cyclopentyl-1,3-dipropylxanthine (DPCPX, 1 microM). Incubation of the tissue with pertussis toxin (PTx, 1 microgram ml-1) did not alter the response to ACh alone but eliminated the synergistic effect of somatostatin. We conclude that nonpigmented epithelial cells of the rabbit ciliary body possess a novel somatostatin receptor whose activation can synergistically potentiate the rise in [Ca2+]i produced by ACh. This potentiation appears to occur via a pertussis-toxin-sensitive pathway, perhaps through Gi. PMID- 9227282 TI - Free radicals in rabbit retina under ocular hyperpressure and functional consequences. AB - Pharmacological experiments have suggested that ocular ischemia, induced by high intraocular pressure in the rabbit, provokes an oxidative stress responsible for functional alteration of the retina. However, the nature of the oxidant chemical species and their mode of generation were not elucidated. The aim of the present studies was to characterize the oxygen-derived free radicals produced during and/or after the hyperpressure period. The technique used was based on electron spin resonance spin trapping analysis of the signals obtained in microdialysates of the retina perfused with the nitrone 5-(diethoxyphosphoryl)-5-methyl-1 pyrroline-N-oxide (DEPMPO). The oxidative stress was also evaluated under ischemia and reperfusion periods by measuring the level of ascorbate in the retina via electron spin resonance detection of the ascorbyl free radical dimethyl sulfoxide (AFR-DMSO) complex. Electroretinograms were recorded to determine the functional consequences of high intraocular pressure and free radical generation. Our results show that superoxide dismutase-inhibitable DEPMPO/hydroxyl radical adducts were generated during the high intraocular pressure period and that the oxidative stress was not increased at reperfusion as assessed by spin trapping and AFR-DMSO measurements. Functional protection provided by free radical scavengers (superoxide dismutase+catalase, TEMPO nitroxide+catalase and dimethylthiourea) against high intraocular pressure induced electroretinogram alteration confirmed these observations. In conclusion, these experiments demonstrate for the first time by direct measurement that oxygen-derived free radicals are produced by the retina during acute ischemia. This generation could be the explanation for electroretinogram alteration. PMID- 9227283 TI - Glial fibrillary acidic protein (GFAP) is synthesized in the early stages of the photoreceptor cell degeneration of the mivit/mivit (vitiligo) mouse. PMID- 9227284 TI - Should the contribution of ACE gene polymorphism to left ventricular hypertrophy be reconsidered? PMID- 9227285 TI - Should we treat hypercholesterolaemia in patients over 65? PMID- 9227286 TI - Joseph Skoda (1805-1881). PMID- 9227287 TI - Patients with suspected myocardial infarction presenting with ST segment depression. PMID- 9227288 TI - Stroke and atrial fibrillation in sick sinus syndrome. PMID- 9227289 TI - Apoptosis in cardiovascular disease. PMID- 9227290 TI - Tricuspid valve endocarditis. PMID- 9227291 TI - Association of angiotensin converting enzyme and angiotensin II type 1 receptor genotypes with left ventricular function and mass in patients with angiographically normal coronary arteries. AB - OBJECTIVE: To analyse the potential association of the angiotensin converting enzyme (ACE) and angiotensin II type 1 receptor (AT1R) gene polymorphisms on left ventricular function and mass in patients with normal coronary arteries. DESIGN: Consecutive sample. SETTING: University hospital. SUBJECTS: 141 consecutive white patients referred for coronary angiography and with angiographically normal coronary arteries. Patients with valvar diseases, cardiomyopathies, or a history of myocardial infarction were excluded. MAIN OUTCOME MEASURES: Left ventricular variables were measured for all patients. The ACE and AT1R genotypes were determined with a polymerase chain reaction based protocol using DNA prepared from white blood cells. A general linear model was used to compare data according to the ACE and to the AT1R genotypes. RESULTS: A strong association was observed between left ventricular mass and systemic hypertension (mean (SD) hypertension: 114 (31) g/m2; no hypertension 98 (23) g/m2; P < 0.003). However, no influence of ACE and AT1R polymorphisms on left ventricular mass was found, regardless of systemic hypertension. The subjects homozygous for the AT1R CC mutation had a significantly lower ejection fraction than those with allele A (AC+AA) (mean (SD) 62(12)% and 68(10)%, respectively, P < 0.05). No synergistic interaction of ACE and AT1R gene polymorphisms on left ventricular function and mass was found. CONCLUSIONS: These data do not support an association of the ACE and AT1R genotypes on left ventricular hypertrophy in white patients with normal coronary arteries. PMID- 9227292 TI - Significance of initial ST segment changes for thrombolytic treatment in first inferior myocardial infarction. AB - OBJECTIVE: To evaluate the benefit to risk ratio of thrombolytic treatment in patients with small inferior acute myocardial infarction (AMI). Controlled studies relating the benefit from thrombolysis with initial electrocardiographic features are scarce and of limited sample size. DESIGN: Retrospective study of 728 patients with a first inferior AMI of six hours' duration from the Intravenous Streptokinase in Acute Myocardial Infarction (ISAM) study comparing streptokinase with placebo stratified by the initial sum ST segment elevation (sigma ST) of 0.8 mV or less and greater than 0.8 mV, and 636 patients from the International Joint Efficacy Comparison of Thrombolytics (INJECT) trial comparing double blind streptokinase with reteplase stratified by either sigma ST or the presence of precordial ST segment depression. RESULTS: ISAM study patients with an sigma ST of greater than 0.8 mV had a significant mortality benefit from streptokinase throughout six years, while those with an sigma ST of 0.8 mV or less showed a trend to higher mortality at six months (6.3% streptokinase v 5.1% placebo). Despite significantly smaller infarcts and fewer clinical complications in patients with an sigma ST of 0.8 mV or less (ISAM and INJECT) or the absence of precordial ST segment depression (INJECT) thrombolytic treatment was associated with higher early mortality than in those with initially larger ST segment deviations. CONCLUSION: Thrombolytic treatment in patients with inferior AMI presenting with larger ST segment deviations is associated with improved survival throughout six years. The risk to benefit ratio, however, in terms of early mortality in patients who have an sigma ST of 0.8 mV or less and no precordial ST segment depression may be unfavourable. PMID- 9227293 TI - Impact of early accelerated dose tissue plasminogen activator on in-hospital patency of the infarcted vessel in patients with acute right ventricular infarction. AB - OBJECTIVE: To assess the efficacy of early accelerated dose tissue plasminogen activator on in-hospital patency of the infarct related artery in patients with inferior myocardial infarction with and without right ventricular involvement. DESIGN: Single centre prospective assessment before discharge of infarct related vessel patency after early thrombolysis. SETTING: Tertiary cardiac referral centre at a university hospital. PATIENTS AND METHODS: 90 consecutive unselected patients with acute myocardial infarction, of whom 35 (39%) had electro cardiographic evidence of right ventricular involvement (ST segment elevation greater than 0.1 mV in right precordial lead V4R), were studied. All patients received accelerated dose tissue plasminogen activator 100 mg within six hours from the onset of symptoms and had control angiography before discharge. MAIN OUTCOME MEASURES: Infarct related coronary artery patency using the Thrombolysis in Myocardial Infarction (TIMI) grading system before discharge. Incidence of prolonged systemic hypotension, sinus bradycardia, complete atrioventricular block, and ventricular tachyarrhythmia during early hospitalisation. RESULTS: Despite aspirin and bolus heparinisation before thrombolysis and high dose heparinisation thereafter for at least 48 hours the infarct related artery was more likely to be occluded (TIMI 0 or 1 flow) in patients with right ventricular involvement than in those without (69 v 29%, P < 0.001), as shown by control angiography performed a mean of 12.8 days after thrombolysis. These findings may be explained, at least in part, by predominant involvement of the proximal right coronary artery (66 v 31%, P < 0.05) and a low cardiac output syndrome, being indirectly reflected by a high incidence of prolonged hypotension (26 v 7%, P = 0.02), bradycardia (34 v 14%, P = 0.03), and complete atrioventricular block (37 v 5%, P = 0.0001). CONCLUSION: Primary angioplasty should be considered as the treatment of choice in patients with acute inferior infarction with right ventricular involvement because of the high failure rate of thrombolysis. PMID- 9227294 TI - Safety of low dose heparin in elective coronary angioplasty. AB - OBJECTIVES: To evaluate the safety of a low dose of heparin in consecutive stable patients undergoing elective percutaneous transluminal coronary angioplasty (PTCA). DESIGN: Open prospective study in a single centre. PATIENTS: 1375 consecutive patients had elective PTCA (1952 lesions: type A 11%, B1 34%, B2 36%, and C 19%). There were no angiographic exclusion criteria. INTERVENTIONS: A bolus of 5000 IU heparin was used as the standard anticoagulation regimen during PTCA. The sheaths were removed immediately after successful completion of the procedure. Prolongation of heparin treatment was left to the operator's discretion. MAIN OUTCOME MEASURES: Procedural success was defined as < 50% residual stenosis without death from any cause, acute myocardial infarction, urgent coronary bypass surgery, or repeat angioplasty within 48 hours for acute recurrent ischaemia; the need for prolonged heparinisation; and the occurrence of puncture site complications. RESULTS: Procedural success without clinical events was achieved in 90% of patients. Mortality was 0.3%; coronary bypass surgery was performed in 1.7% of the procedures. The rate of myocardial infarction was 3.3%; repeat angioplasty within 48 hours was carried out in 0.7% of patients. A total of 89.1% of the patients were treated according to the protocol. Prolonged treatment with heparin was considered necessary in 123 patients (8.9%). Repeat angioplasty for abrupt closure was performed in two patients shortly after sheath removal and in two during prolonged heparinisation. Puncture site complications occurred in 2.1% of patients (low dose heparin 1.9% and prolonged heparinisation 4.9%). CONCLUSION: Elective PTCA can be safely performed using a low dose of heparin, with a negligible risk for subacute closure. Low dose heparin may reduce the incidence of puncture site complications, shorten hospitalisation, and enable out-patient angioplasty. PMID- 9227295 TI - Effect of oral aminophylline in patients with angina and normal coronary arteriograms (cardiac syndrome X). AB - BACKGROUND: Patients with syndrome X (exertional angina, positive exercise test, normal coronary arteriogram) have an altered perception of cardiac pain. This symptom may arise from increased sensitivity to adenosine. Previous studies suggest that intravenous aminophylline (an adenosine receptor blocker) improves exercise tolerance in patients with this disorder. OBJECTIVE: To examine the efficacy of oral aminophylline in syndrome X. METHODS: 13 patients (11 women and two men, mean (SD) 54 (6) years) with syndrome X were studied. Patients were randomised in a double blind crossover study to receive either oral aminophylline or placebo for three weeks. All patients underwent symptom limited exercise testing and ambulatory electrocardiography at the end of each three week period. RESULTS: 10 patients completed the study. The time to angina during exercise testing in patients who were given aminophylline was longer than for the placebo group (mean (SD) 632 (202) seconds v 522 (264) seconds, P = 0.004). Peak exercise ST depression did not differ significantly between patients who received aminophylline and those administered placebo (mean (SD) -1.9 (0.7) mm v -1.5 (0.8) mm). Six patients taking aminophylline reported a reduction in the total number of episodes of chest pain during the three weeks, but the frequency and duration of ST segment depression during Holter monitoring was unchanged. CONCLUSION: Oral aminophylline has a favourable effect on exercise induced chest pain threshold in patients with syndrome X. The disparate effects on symptoms and ST segment changes are intriguing and further study is warranted. PMID- 9227296 TI - The role of signal averaged P wave duration and serum magnesium as a combined predictor of atrial fibrillation after elective coronary artery bypass surgery. AB - OBJECTIVE: To investigate the role of low serum magnesium as a trigger for atrial fibrillation in patients with a substrate for the arrhythmia (assessed by signal averaged P wave duration). DESIGN: A case-control study. SETTING: A regional referral cardiac centre. PATIENTS AND INTERVENTIONS: 105 consecutive patients undergoing elective coronary artery bypass surgery had signal averaged P wave recordings before operation. Serum electrolytes were analysed preoperatively and on days 1, 2, and 5 after surgery. MAIN OUTCOME MEASURES: Any episode of electrocardiographically recorded atrial fibrillation was taken as a study end point. RESULTS: Of 102 patients discharged, 27 (26%) had documented episodes of atrial fibrillation at a mean of 2.7 days after surgery. A combination of P wave duration > 155 ms and serum magnesium on the first postoperative day of < 0.7 mmol/l had a sensitivity of 75% and specificity of 80% for predicting atrial fibrillation. Duration of hospital stay (7.9 v 6.8 days) was longer in the atrial fibrillation group (P < 0.01). Stepwise regression showed age, serum magnesium < 0.7 mmol/l on the first postoperative day (both P < 0.001), angiotensin converting enzyme inhibitor withdrawal (P < 0.02), and signal averaged P wave duration (P = 0.04) to be independent predictors. CONCLUSIONS: The combination of signal averaged P wave duration and low serum magnesium on the first postoperative day identified the majority of patients with atrial fibrillation after coronary artery bypass surgery. Early identification and pharmacological treatment for selected patients may reduce the incidence of postoperative atrial fibrillation. PMID- 9227297 TI - Sympathetic reinnervation and heart rate variability after cardiac transplantation. AB - BACKGROUND: Heart rate variability is thought to measure autonomic modulation, but the relation has never been demonstrated directly in humans. AIM: To test the hypothesis that increased low frequency heart rate variability reflects sympathetic reinnervation after cardiac transplantation. PATIENTS: 24 cardiac transplant recipients at the time of routine surveillance coronary angiography two or more years after cardiac transplantation, and 10 controls with normal coronary arteries undergoing angiography for investigation of chest pain. SETTING: Regional cardiothoracic centre. METHODS: Sympathetic effector function at the sinus node was assessed by measuring the fall in cycle length for two minutes after injection of tyramine to the artery supplying the sinus node. Heart rate variability was measured from three-minute RR interval sequences at rest, during metronomic respiration, and before and after atropine. RESULTS: The logarithm of the low frequency component of heart rate variability during metronomic respiration was linearly related to the logarithm of the change in cycle length after injection of tyramine (R2 = 0.28, P = 0.007). Absolute units more accurately reflected sympathetic effector function than did normalised units or the ratio of low frequency to high frequency. Atropine did not affect high frequency heart rate variability in transplant recipients. CONCLUSIONS: The low frequency component of heart rate variability is directly related to sympathetic reinnervation to the sinus node. PMID- 9227298 TI - Measuring QT dispersion: man versus machine. AB - OBJECTIVE: To compare manual and computer automated techniques for measuring QT dispersion. DESIGN: Assessment of the ability of manual and automatic measurements of QT dispersion to discriminate between a normal group and two cardiac groups. SUBJECTS: 12 simultaneous electrocardiogram leads were recorded from 25 healthy volunteers, 25 subjects after myocardial infarction, and 25 with cardiac arrhythmias. MAIN OUTCOME MEASURES: For each subject, QT dispersion was measured as the difference between the maximum and minimum QT from all 12 leads and separately for only those leads with T amplitudes of > 100 microV and for those > 250 microV. RESULTS: Manual QT dispersion (T > 100 microV) was greater (P < 0.02) in the arrhythmia patients (mean (SD), 45 (21) ms), but not the infarction patients (54 (36) ms), than in the normal subjects (39 (13) ms). There were no significant differences when all T waves were included. QT dispersion was significantly reduced by an average of 30% when T waves < 100 microV were excluded, and by 51% when those < 250 microV were excluded. Automatic techniques gave different measurements for dispersion in comparison with manual measurements. Three of the four automatic techniques detected significant differences between normal and both patient groups when no leads were excluded (P < 0.01) as well as when T waves < 100 microV were excluded (with increased significance, P < 0.002). CONCLUSIONS: Measurements of QT dispersion from small T waves increases measurement variability and reduces the potential for detecting clinical differences. Automatic measurement of QT dispersion gives different results from manual measurement, but can satisfactorily discriminate between normal and abnormal groups with good quality electrocardiograms. PMID- 9227299 TI - QT interval as a cardiac risk factor in a middle aged population. AB - OBJECTIVE: To evaluate the value of QT interval as a cardiac risk factor in middle aged people. METHODS: The association between QT interval and cardiac risk factors and mortality in a middle aged Finnish population of 5598 men and 5119 women was evaluated over a 23 year follow up. To adjust the QT interval confidently for heart rate, a nomogram was constructed from the baseline electrocardiograms separately for men and women. RESULTS: Nomogram-corrected QT interval (QTNc) prolongation was associated with elevated blood pressure and signs of cardiovascular disease; QTNc shortening was associated with smoking. Over 10% prolongation of QTNc predicted death in men with heart disease: adjusted relative risk (RR) was 2.17 (95% confidence interval 0.67-7.45) for sudden death; 2.12 (1.25-3.59) for total cardiovascular mortality; and 1.92 (1.23-3.00) for all cause mortality. In healthy men the increase in RR was not significant: sudden death, 1.48 (0.67-3.25); total cardiovascular mortality, 1.25 (0.92-1.70); all cause mortality, 1.21 (0.96-1.53). However, healthy men with long QTNc in the lowest heart rate quartile exhibited an RR of 2.75 (1.00-7.40) for sudden death. Over 10% shortened QTNc predicted cardiovascular death in men with heart disease who smoked; RR 3.72 (1.45-9.54). Non-smoking men with short QTNc had low mortality risks irrespective of possible signs of cardiovascular disease. The trends in mortality risks were similar but weaker for women. CONCLUSIONS: In a middle aged population, prolonged QT interval predicts cardiac mortality in men with signs of cardiovascular disease. In women and healthy men this risk is weak and may reflect subclinical heart disease. A shortened QT interval predicts death in men with heart disease who smoke. PMID- 9227300 TI - Idiopathic dilated cardiomyopathy: familial prevalence and HLA distribution. AB - OBJECTIVES: To compare HLA distribution in familial and non-familial dilated cardiomyopathy, because a serum marker that could identify families at risk of developing dilated cardiomyopathy should be of use in screening for the disease. PATIENTS: 100 patients with dilated cardiomyopathy. METHODS: 200 first degree relatives from 56 of the proband families were screened for dilated cardiomyopathy by echocardiography. The HLA profile of the patients with dilated cardiomyopathy, as well as of the familial and non-familial subgroups, was compared with that of 9000 normal controls. RESULTS: The familial prevalence of dilated cardiomyopathy in this patient group was "definite" in 14 of 56 (25%) and "possible" in 25 of 56 (45%). The HLA-DR4 frequency in the 100 patients with dilated cardiomyopathy was similar to that in the 9000 controls (39% v 32%). However, the DR4 subtype was significantly more common in the 25 probands with a familial tendency to dilated cardiomyopathy than in the 31 probands with non familial dilated cardiomyopathy (68% v 32%; P < 0.05). CONCLUSIONS: The present finding supports an HLA linked predisposition to familial dilated cardiomyopathy. The HLA type DR4 was significantly more common in familial than in non-familial cases. The DR4 halotype was associated with two thirds of the families at risk for dilated cardiomyopathy. PMID- 9227301 TI - A study comparing VVI and DDI pacing in elderly patients with carotid sinus syndrome. AB - OBJECTIVE: To determine whether single chamber ventricular demand (VVI) pacing is adequate for elderly patients with carotid sinus syndrome. DESIGN: Prospective double blind randomised cross over study. SETTING: Tertiary referral centre. PATIENTS: 30 consecutive patients aged over 60 years with carotid sinus syndrome referred for cardiac pacing. INTERVENTION: Patients underwent dual chamber pacemaker implantation and were then randomised to two three-month periods of VVI and DDI pacing. MAIN OUTCOME MEASURES: Responses to cardiovascular tests (vasodepression during carotid sinus massage, pacemaker effect, postural blood pressure measurements, and response to head up tilt), and symptoms. RESULTS: 11 patients developed profound hypotension during upright carotid sinus massage while pacing VVI compared with only two while pacing DDI. The upright pacemaker effect was greater in VVI (VVI, -31 (SD 19) mm Hg v DDI, -4 (12) mm Hg; P < 0.001). Postural blood pressure measurements and responses to head up tilt did not vary. Eleven patients were unable to tolerate VVI pacing and had to be withdrawn early from this limb of the study (group A). Fourteen of the remainder completed diary cards and did not express a preference (group B). No patient preferred VVI. Group A patients were older (group A, 78 (6) years v group B, 70 (9) years; P < 0.05), were more likely to be female (group A, 73% v group B, 14%; P < 0.01), and were more likely to have orthostatic hypotension while pacing DDI (group A, 46% v group B, 0%; P < 0.01). Group A and B patients could not be differentiated by other prepacing clinical or haemodynamic variables. CONCLUSIONS: Elderly patients with carotid sinus syndrome are likely to develop symptomatic hypotension following VVI pacing. The optimum pacing mode for individual patients cannot be predicted by simple cardiovascular tests before pacing. PMID- 9227303 TI - Percutaneous balloon mitral commissurotomy during pregnancy. AB - OBJECTIVE: To evaluate the effectiveness and safety of percutaneous balloon mitral commissurotomy for the treatment of pregnant women with severe mitral stenosis over a period of six years. DESIGN: Analysis of clinical, haemodynamic, and echocardiographic data before and immediately after the procedure, the pregnancy outcome, and the fate of newborn babies. SETTING: Academic cardiovascular centre in Monastir, Tunisia. PATIENTS: 44 pregnant patients who underwent percutaneous transvenous dilatation of the mitral valve between January 1990 and February 1996. Grade 2 mitral regurgitation was present in two patients and densely calcific valves in three (7%). RESULTS: Commissurotomy was successfully achieved in all cases. The total mean (SD) duration of teh procedure was 72 (18) minutes and that of fluoroscopy 16 (7) minutes. Left atrial pressure decreased from 28 (10) to 14 (7) mm Hg, mitral pressure gradient fell from 22 (8) to 5 (3) mm Hg. Cardiac output increased from 4.8 (1.1) to 6.3 (1.2) l/min and Gorlin mitral valve area from 0.96 (0.21) to 2.4 (0.4) cm2 (all P < < 0.001). Cross sectional echocardiographic mitral valve area increased from 1.07 (0.21) to 2.32 (0.36) cm2. There were no maternal or fetal deaths. Complications included a grade 4 mitral regurgitation in one patient that required early valve replacement. All patients delivered at full term, 42 vaginally and two (5%) by caesarean section; 41 babies were normal and three whose mothers had the procedure near term were relatively hypotrophic. At a mean follow up of 28 (12) months (range 2 to 26) all children had normal growth. CONCLUSIONS: During pregnancy, balloon mitral commissurotomy is the treatment of choice of severe pliable mitral stenosis in patients who are refractory to medical treatment. PMID- 9227304 TI - John Honour, Sir Thomas Lewis's personal laboratory assistant. PMID- 9227302 TI - Transthoracic Doppler echocardiographic analysis of phasic coronary blood flow velocity in hypertrophic cardiomyopathy. AB - OBJECTIVE: To use transthoracic Doppler echocardiography to assess coronary blood flow non-invasively in patients with hypertrophic cardiomyopathy. DESIGN: High frequency transthoracic Doppler echocardiography was used to assess resting phasic coronary velocity patterns in patients with hypertrophic cardiomyopathy and to define the relation between coronary flow patterns and clinical, echocardiographic, and haemodynamic manifestations of this condition. SETTING: A tertiary referral cardiothoracic centre. METHODS: Fifteen patients (10 men and five women, mean (SD) age 49 (10.3) years) with asymmetric hypertrophic cardiomyopathy underwent high frequency (5 MHz) transthoracic Doppler echocardiographic assessment of the left anterior descending coronary artery. In addition, standard two dimensional echocardiography was performed. The results were compared with 16 normal participants (nine men and seven women, mean age 61.2 (10.7) years) who had no evidence of cardiac disease. RESULTS: Biphasic diastolic predominant coronary artery blood velocity profiles were obtained in all patients and controls. Systolic peak blood velocity and velocity time integral were significantly reduced in the hypertrophic cardiomyopathy group compared with controls (11.3 (15.8) cm/s and 1.09 (1.78) cm v 20.5 (13.1) cm/s and 4.23 (2.80) cm, respectively, P < 0.05). A reversed pattern of systolic blood flow velocity was found in three patients with severe anterior wall and septal hypertrophy. During diastole there was prolongation of the diastolic acceleration (203 (53) ms v 110 (60) ms in controls, P < 0.05) and deceleration times (487 (200) ms v 210 (90) ms in controls, P < 0.05). There was no significant difference between those with and without symptoms or a left ventricular outflow tract gradient. CONCLUSIONS: Patients with hypertrophic cardiomyopathy have abnormal systolic and diastolic coronary flow profiles at rest when measured by transthoracic echocardiography. PMID- 9227305 TI - Papillary endothelial hyperplasia in a TEC coronary atherectomy specimen. AB - Angiography in a 37 year old female with a three week history of typical crescendo angina found an 80% stenosis of the proximal left anterior descending (LAD) artery. The patient underwent percutaneous transluminal coronary angioplasty involving TEC artherectomy of the LAD artery. The specimen removed by atherectomy was found to have the appearance of papillary endothelial hyperplasia. This is an unusual histological diagnosis that occurs in association with thrombus. It is rarely found within arterial vessels and has not been reported in a coronary artery. Papillary endothelial hyperplasia is now thought to be a form of organising thrombus, probably dependent on the production of basic fibroblast growth factor by the endothelium. PMID- 9227306 TI - Percutaneous transhepatic dual chamber pacing in children with Fontan circulation. AB - Permanent pacing is often required following the Fontan operation and is usually performed epicardially as there is no direct access to the ventricle from the systemic veins. Dual chamber endocardial pacing was achieved by the transhepatic approach in two children with Fontan circulation. The patients were a 7 year old boy with left atrial isomerism, single ventricle with pulmonary stenosis, interrupted inferior vena caval vein with azygous continuation, and direct drainage of the hepatic veins to the right sided atrium, and a 6 year old girl with tricuspid atresia. This approach to endocardial pacemaker implantation is potentially of considerable value in patients who do not have direct access to the ventricle from the systemic veins. PMID- 9227307 TI - Coronary involvement in the Churg-Strauss syndrome. AB - The Churg-Strauss syndrome (CSS) is a rare systemic disease characterised by vasculitis and peripheral eosinophilia in patients with an atopic constitution. Cardiac involvement is an important cause of morbidity and mortality yet coronary involvement is very rarely documented. We report the case of a 38 year old man presenting with fulminant heart failure. Coronary arteriography demonstrated extensive focal vasculopathy consistent with vasculitis. The diagnosis of CSS was established based upon the classical diagnostic criteria and corticosteroid treatment resulted in a spectacular remission of disease activity. PMID- 9227308 TI - Occlusion of persistent left superior vena cava to unroofed coronary sinus using vena cava filter and coils. AB - A 48 year old female with complex cyanotic heart disease and pulmonary hypertension was partly cyanosed because of a persistent left superior vena cava draining into an unroofed coronary sinus. The left superior vena cava, which measured 22 mm in diameter, was successfully occluded with a Gunther Tulip Vena Cava Mreye Filter which acted as a barrier for embolisation coils. PMID- 9227309 TI - Intractable recurrent ventricular tachycardia in dilated cardiomyopathy controlled by a vasodilating beta blocker. PMID- 9227310 TI - Acute hepatitis complicating parenteral amiodarone does not preclude subsequent oral therapy. PMID- 9227311 TI - Endothelial dysfunction in the absence of coronary atheroma causing Prinzmetal's angina. PMID- 9227312 TI - Epitope specificity of IgE antibodies to a major allergen (Cry j 1) of Japanese cedar pollen in sera of humans and monkeys with pollinosis. AB - Japanese cedar (Cryptomeria japonica) pollinosis has been reported to occur naturally in Japanese monkeys (Macaca fuscata) as well as humans. Using monoclonal antibodies (mAb) specific to Cry j 1, a major allergen in Japanese cedar pollen, we identified five independent epitopes (EP-1 to EP-5) on the molecule. The epitopes recognized by IgE antibodies in the sera of humans and monkeys with the pollinosis were analysed by an IgE enzyme-linked immunosorbent assay inhibition method with these mAb. In human patients, the mAb to EP-1 strongly blocked the binding of IgE antibodies in all patients' sera to Cry j 1. The reaction patterns of IgE antibodies in monkeys, however, varied among the troops of monkeys. In some troops, the mAb to EP-1 showed a blocking pattern similar to that for human patients. In other troops, mAb to EP-4 and EP-5 blocked binding of IgE. These results indicate that some, but not all, monkeys have antibody responses to the major allergen similar to those of humans. PMID- 9227313 TI - Immunoglobulin response to intrapulmonary immunization of asthmatics. AB - Atopic asthmatics, compared to non-atopic individuals, exhibit an increased amount of serum antigen-specific IgE and IgG4 antibody directed toward many aeroallergens. We tested the hypothesis that this difference between atopics and non-atopics extends to the response to intrapulmonary deposition of a neoantigen, keyhole limpets haemocyanin (KLH). We immunized nine atopic asthmatics and nine non-atopic controls with 500 micrograms KLH instilled into a subsegment of the lingula and examined serum anti-KLH, IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, and IgM and specific antibody production by peripheral blood mononuclear cells for 25 days. We also determined specific antibody in bronchoalveolar lavage fluid (BALF) in both the immunized and a non-immunized lobe 11 days after immunization. We found specific serum antibody in all immunized subjects with no difference between atopics and normals in the amount or kinetics of anti-KLH IgG1, IgG2, IgG3, IgA1, IgA2 and IgM. However, the atopics exhibited more anti-KLH IgG4 than the normal controls. Specific anti-KLH antibody-producing cells were detected in peripheral blood in most subjects at day 8 to 12 after immunization with no difference between atopics and normals. Specific IgA1, IgA2, IgG1 and IgM antibodies were detected in BALF from the immunized lobes but not from the non immunized lobes of both groups of subjects with no difference between atopics and normals. We conclude that atopic asthmatics respond to intrapulmonary KLH with more serum anti-KLH IgG4 than normal controls, consistent with a bias toward a Th2 response to intrapulmonary exposure to antigen. PMID- 9227314 TI - Adoptive transfer of allergen-specific CD4+ T cells induces airway inflammation and hyperresponsiveness in brown-Norway rats. AB - Following allergen exposure, sensitized Brown-Norway rats develop airway hyperresponsiveness (AHR) and eosinophilic inflammation together with an increase in activated T cells (CD25+) in the airways. We tested the hypothesis that CD4+ T cells are involved directly in the acquisition of AHR. Spleen T cells from animals that were injected intraperitoneally on three consecutive days with ovalbumin/Al(OH)3, showed a dose-dependent proliferative response in vitro to ovalbumin, but not to bovine serum albumin, as measured by [3H]thymidine uptake. For total T-cell transfer, spleen cells obtained from donor rats 4 days after sensitization were depleted of adherent cells by a nylon wool column separation. CD4+ and CD8+ T cells were purified by immunomagnetic beads cell separation. Recipient naive rats were injected intravenously with 50 x 10(6) total T cells, 20 x 10(6) and 5 x 10(6) CD4+ cells, and 5 x 10(6) CD8+ cells, and were exposed to ovalbumin aerosol 24 hr afterwards. After a further 24 hr, airway responsiveness to acetylcholine (ACh) was measured and provocative concentration (PC) values PC100, PC200 and PC300) (the ACh concentration needed to achieve 100, 200 and 300% increase in lung resistance above baseline) were calculated. Airway responsiveness was significantly increased in recipients of sensitized total T cells compared with recipients of cells from saline-injected donor rats (P < 0.05). There were significantly increased eosinophil major basic protein (MBP)+ cell counts/mm2 in airway submucosal tissue in the hyperreactive rats and a significant correlation was found between the number of MBP+ cells and PC100 (r = 0.75; P < 0.03) in recipients of sensitized total T cells. Purified CD4+ T cells from sensitized donors induced AHR in naive recipients (P < 0.05), while sensitized CD8+ and naive CD4+ cells failed to do so. Our data indicate that T cells may induce AHR through an eosinophilic airway inflammation and that CD4+ T cells may have a direct effect in this process in Brown-Norway rats. PMID- 9227315 TI - Cytotoxic T-lymphocyte interaction with fibronectin and vitronectin: activated adhesion and cosignalling. AB - Stimulation of cloned cytotoxic T lymphocytes (CTL) with anti-T-cell receptor (TCR) monoclonal antibody (mAb) in solution resulted in rapid and sustained activation of adhesion to immobilized fibronectin (FN) but did not initiate degranulation. Addition of a second antibody (Ab) to further cross-link the TCR substantially increased the level of adhesion and also activated degranulation, as measured by release of serine esterase, in the presence of immobilized FN but not in its absence. Thus, binding to FN can provide a costimulatory signal to activate degranulation. TCR cross-linking also activated CD8-dependent adhesion to class I, and CD8 provided a costimulatory signal upon binding to class I. However, the requirements for activating adhesion and generating the costimulatory signal differed significantly for FN versus class I ligand, suggesting that these two receptor-ligand systems do not share a common mechanism of action. Co-immobilizing FN and alloantigen resulted in increased serine esterase release in comparison with that stimulated by antigen alone, and required the FN and class I be on the same surface. Peptide and antibody blocking demonstrated that CTL binding to FN, and to vitronectin (VN), was mediated by the alpha V beta 3 vitronectin receptor (VNR). Thus, VNR is activated by a signal from the TCR to mediate adhesion to FN or VN, and delivers a costimulatory signal for degranulation via a different mechanism than costimulation by CD8 binding to class I. PMID- 9227316 TI - Effect of lisofylline and pentoxifylline on the bacterial-stimulated production of TNF-alpha, IL-1 beta IL-10 by human leucocytes. AB - The present study concerns the effect of the xanthine derivates lisofylline (LSF) and pentoxifylline (PTX) on the production of pro-inflammatory cytokines tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) and the de activating cytokine interleukin-10 (IL-10) by human leucocytes during stimulation with lipopolysaccharide (LPS), heat-killed Gram-negative bacteria (GNB) or Gram positive bacteria (GPB). The production of TNF-alpha and IL-1 beta by leucocytes stimulated with LPS, Haemophilus influenzae type b (Hib) or Streptococcus pneumoniae was inhibited by both drugs. The production of IL-10 by leucocytes stimulated with LPS and Hib was inhibited by both xanthine derivates only at 48 hr. However, incubation of leucocytes with S. pneumoniae in the presence of LSF or PTX stimulated the production of IL-10 about four- and twofold at 24 hr and 48 hr, respectively. In all instances, the extent of inhibition or enhancement of cytokine production by LSF or PTX was equal. The divergent effects of xanthine derivates on the IL-10 production indicate the existence of distinct intracellular pathways depending on whether leucocytes are stimulated by GPB or GNB. PMID- 9227318 TI - Differential expression of CD32 isoforms following alloactivation of human T cells. AB - Receptors for the Fc region of immunoglobulin G (IgG) (Fc gamma Rs) exist in three main forms: membrane bound, soluble and cytoplasmic. The function of cytoplasmic Fc gamma Rs is poorly understood. We have previously demonstrated cytoplasmic Fc gamma RII (cCD32) within most normal human peripheral blood lymphocytes (PBL), including T cells. In this study we have investigated the hypothesis that following lymphocyte activation, up-regulation of cCD32 occurs, resulting in increased expression at the cell surface. Normal PBL were activated in vitro using a two-way mixed lymphocyte reaction (MLR) and expression of CD32 monitored by flow cytometry and by immunoperoxidase staining using specific monoclonal antibodies and aggregated mouse IgG subclasses. Furthermore, we designed oligonucleotide probes specific for the three main isoforms of CD32 and looked for changes in mRNA expression throughout the MLR using an in situ hybridization technique. Increased surface expression of CD32 was found on both activated human T and B lymphocytes, but this was found only in the early stages of the MLR, on days 3 and 4, and was virtually absent by day 7. An inverse relationship between cell surface expression of CD32 and mRNA for the IIb isoforms was noted with strong mRNA expression for IIb isoforms occurring in the later stages of the MLR (days 6-7) when interleukin-2R (IL-2R)-positive T cells were predominant. A soluble IgG binding factor (soluble CD32?) was also detected in the MLR culture supernatant. These observations provide support for the hypothesis that synthesis of IIb isoforms of CD32 occurs following alloantigen activation of human T lymphocytes. PMID- 9227317 TI - Inhibition of human interleukin-12 production by pentoxifylline. AB - Pharmacological control of interleukin-12 (IL-12) production may be a key therapeutic strategy for modulating immunological diseases dominated by type-1 cytokine responses. In this study, we investigated the effects of pentoxifylline on the production of IL-12 by human blood mononuclear cells and primary human monocytes stimulated with heat-killed Staphylococcus aureus Cowan strain I (SAC) or lipopolysaccharide (LPS). Pentoxifylline potently suppressed production of IL 12 in a concentration-dependent manner. In these same experiments, tumour necrosis factor-alpha (TNF-alpha) production was inhibited and IL-10 and prostaglandin E2 (PGE2) production was enhanced by treatment with pentoxifylline. Suppression of IL-12 production by pentoxifylline was found to be independent of several known endogenous inhibitors of IL-12, such as IL-10, transforming growth factor-beta (TGF-beta), IL-4 and PGE2. RNase protection assays revealed that pentoxifylline inhibited accumulation of both IL-12 p40 and p35 mRNA, suggesting a predominant mRNA locus for pentoxifylline-induced IL-12 inhibition. Low levels of pentoxifylline added to the suppression of IL-12 production by suboptimal inhibiting doses of dexamethasone, suggesting that this drug combination may have therapeutic utility. These results provide a firm rationale for the use of pentoxifylline in clinical trials of immunological disorders characterized by inappropriate type-1 immune responses. PMID- 9227319 TI - Enhancement of antigen-presenting ability of B lymphoma cells by partial inhibition of protein synthesis through inducing B7-1 expression. AB - During the investigation of the role of protein synthesis in antigen-presenting cell (APC) function of A20-HL B lymphoma cells, we found that partial inhibition of protein synthesis enhanced their APC function. The treatment of A20-HL cells with 0.313-2.5 microM emetine, an irreversible inhibitor of protein synthesis, decreased protein synthesis by 60-70%, and enhanced their APC function to stimulate I-Ad/OVA323-339-specific T cells to produce interleukin-2 in response to ovalbumin (OVA). The emetine-treated and paraformaldehyde-fixed A20-HL cells required only 20 nM OVA323-339 peptide to stimulate the T cells, whereas those untreated and fixed required 200 nM peptide. This enhancement of APC function was mostly because of the induction of B7-1 expression on A20-HL cells by the emetine treatment, since B7-1 molecules were detected on the emetine-treated A20-HL cells, but only negligibly, if at all, on the untreated cells, and an anti-B7-1 monoclonal antibody, 1G10, inhibited the enhanced APC function of the emetine treated A20-HL cells. The emetine-treatment also increased B7-1 mRNA expression in A20-HL cells, suggesting that the induction of B7-1 expression was due to the increase in the accumulation of mRNA and the translation with residual ability to synthesize protein. Thus, partial inhibition of protein synthesis in A20-HL cells increases B7-1 mRNA accumulation and its expression on the cell surface, which results in the enhancement of their APC function. PMID- 9227321 TI - Correlation between natural killer cell activation in the bone marrow and haemopoietic dysfunction following cytomegalovirus infection of mice. AB - We describe here the activation of natural killer (NK) cells in the bone marrows and spleens of mice infected with murine cytomegalovirus (MCMV). NK activity at these sites peaked at day 2 to 3 post-infection (p.i.) and declined between days 6 and 10 p.i. in BALB/c and C57BL/6 mice. In BALB/c mice, the increases in NK activity coincided with depletion of colony-forming units of the granulocyte monocyte lineage (CFU-GM) from the marrow. CFU-GM depletion in MCMV-infected C57BL/6 mice was less severe, despite the presence of activated NK cells in the marrow. Treatment of BALB/c mice with anti-asialo GM1 prior to MCMV infection resulted in less severe CFU-GM depletion at day 2 p.i. than infection with MCMV alone. When homozygous C57BL/6 or CBA/CaH bg/bg mice were infected with MCMV, depletion of marrow CFU-GM was more severe than in their heterozygous littermates. Finally, we observed some inhibition of colony formation when marrow cells from MCMV-infected and uninfected BALB/c donors were mixed and incubated prior to the CFU-GM assay. These results suggest that activated NK cells may contribute to depletion of haemopoietic cells soon after MCMV infection of BALB/c mice, but may limit the loss of these cells in C57BL/6 and CBA/CaH mice. PMID- 9227320 TI - Autologous killing by a population of intermediate T-cell receptor cells and its NK1.1+ and NK1.1- subsets, using Fas ligand/Fas molecules. AB - Self-reactive clones, estimated by anti-V beta monoclonal antibodies (mAb) in conjunction with the Mls system, are confined to a population of intermediate (int) T-cell receptor (TCR) (or CD3) cells (i.e. TCRint cells), but are not found among TCRhigh cells. The next questions to be answered are whether autologous killing is confined to TCRint cells and how such killing is mediated. In this study, 51Cr-labelled thymocytes of syngeneic or allogeneic origin were used as target cells (4-hr assay). When liver and splenic mononuclear cells (MNC) obtained from B6 mice were used as effector cells, prominent autologous killing was seen in liver MNC, but not splenic MNC. Such killing was not seen when thymocytes from B6-lpr/lpr mice (i.e. Fas-) were used as target cells, nor when liver MNC from MRL-gld/gld mice (i.e. Fas ligand-) were used as effector cells (target thymocytes of MRL(-)+/+ mice). Cell separation experiments using a cell sorter revealed that autologous killing was mediated for the most part by CD3int cells, while allogeneic killing was mediated entirely by natural killer (NK) cells, TCRint cells and TCRhigh cells. Among CD3int cells, the NK1.1+ subset (i.e. NK1.1+ T cells) manifested a higher level of autologous killing than did the NK1.1- subset. Consistent with the results of a functional assay, it was found by reverse-transcription-polymerase chain reaction (RT-PCR) assay that CD3int cells among liver MNC showed the expression of Fas ligand mRNA, while thymocytes expressed Fas mRNA. When class I major histocompatibility complex (MHC)- thymocytes (from beta 2-microglobulin-deficient mice) were used as target cells, NK cells, but not CD3int cells, showed potent cytotoxicity. These results suggest that autologous killing is a major function of TCRint cells with self reactivity, and that such killing is mediated by means of Fas ligand/Fas molecules. PMID- 9227322 TI - Heterogeneity of cytokine functions in HIV infection. AB - Immunological responses, especially cytokines, play important roles in determining the persistence of infectious agents in chronic diseases. Th1 responses enhance cellular immunity to control infection whereas Th2 immune responses down-regulate these effector immune responses. It has been suggested that the Th1 to Th2 switch is involved in human immunodeficiency virus (HIV) disease progression. We studied the regulatory role of interleukin-4 (IL-4; Th2 response) on interferon-gamma (IFN-gamma; Th1 response) in HIV infection and its role in the generation of HIV-specific cytotoxic T lymphocytes (CTL) in an in vitro system. Forty HIV-infected, asymptomatic individuals and 20 HIV seronegative individuals were included in this study. Peripheral blood mononuclear cells were stimulated with phytohaemagglutinin and tetanus toxoid in the presence or absence of IL-4 to determine the effect of IL-4 on IFN-gamma production and HIV-Env-specific CTL activity. IL-4 showed a dual effect on IFN gamma production in HIV patients. IL-4 down-regulated IFN-gamma production in HIV seronegative individuals and in 55% of HIV patients whereas it stimulated IFN gamma production in 45% of HIV patients. IL-4 increased HIV-Env-specific CTL activity in five of seven patients of the latter group. IL-4 has multiple biological activities, e.g. IL-4 inhibits IFN-gamma production as well as stimulates CTL generation which in turn produces IFN-gamma. Understanding the biological significance of these interactions is of importance for immunotherapeutic approaches against HIV infection. PMID- 9227323 TI - Infection of C3HeB/FeJ mice with the docile strain of lymphocytic choriomeningitis virus induces autoantibodies specific for erythrocyte Band 3. AB - C3HeB/FeJ mice infected with the docile strain of lymphocytic choriomeningitis virus (LCMV-d) develop a persistent infection with a transient haemolytic anaemia. Immunoglobulin can be eluted from the red blood cells (RBC) of these mice but it cannot be detected on the RBC by a conventional antiglobulin test. The present study demonstrates that RBC from such mice bear erythrocyte autoantibodies which are predominantly of the IgG2a subclass, with lower levels of autoantibodies of the IgG1, IgG2b and IgG3 subclasses. To identify the target antigen the autoantibodies were eluted from the RBC of LCMV-infected mice. The eluted autoantibody bound to intact normal RBC and precipitated a 105000 MW component that corresponds to murine Band 3 protein. A monoclonal antibody derived from mice infected with LCMV-d also precipitated mouse Band 3, and reacted specifically by enzyme-linked immunosorbent assay against a purified preparation of Band 3. This study has shown that in C3H mice infected with LCMV-d which develop autoimmune haemolytic anaemia, the target autoantigen is erythrocyte membrane Band 3. PMID- 9227324 TI - The anti-erythrocyte autoimmune response of NZB mice. Identification of two distinct autoantigens. AB - With age, New Zealand black (NZB) mice spontaneously develop anti-mouse red blood cell (RBC) autoantibodies resulting in the development of autoimmune haemolytic anemia (AIHA). Previously, we characterized a panel of monoclonal autoantibodies derived from unimmunized, adult NZB mice. One of these antibodies (G8) was shown to be pathogenic, inducing AIHA in a non-autoimmune-prone mouse strain (BALB/c). Using G8, and two other antibodies from our panel, we have characterized two distinct autoantigens on the surface of mouse RBCs. The autoantigen, historically referred to as antigen X (AgX), was found to be partially hidden on the surface of the mouse RBC because glycosidase treatment or mild digestion with proteinase K resulted in increased reactivity with autoantibodies. One of the monoclonal antibodies (3H5G1) was found to immunoprecipitate a 110,000 MW protein identified as the erythrocyte anion transporter (band 3) whereas the pathogenic antibody (G8) as well as a third monoclonal antibody (2E6m) were shown to immunoprecipitate a 60,000 MW protein that was not reactive with the anti-band 3 serum. Finally, we show that the autoantigen recognized by G8 is expressed on differentiated mouse erythroleukaemia (MEL) cells. The results suggest that a protein distinct from band 3 can serve as a target for AIHA in NZB mice. PMID- 9227325 TI - Inhibitory effects of interleukin-10 on synovial cells of rheumatoid arthritis. AB - This paper describes the immunoregulatory effects of interleukin-10 (IL-10) on synovial cells in vitro. Synovial cells were cultured with IL-10 in the presence or absence of various cytokines. Following incubation, the costimulatory molecule expression on synovial cells and cytokine production in culture supernatants were analysed by an indirect immunofluorescence method and enzyme-linked immunosorbent assay, respectively. We also examined the effect of IL-10 on the function of synovial cells as antigen-presenting cells (APC). Synovial cells spontaneously express several kinds of costimulatory molecule and produce various kinds of cytokines. Stimulation of synovial cells with interferon-gamma (IFN-gamma), IL-1 beta, or 12-O-tetradecanoyl phorbol 13-acetate (TPA) markedly enhanced the expression of costimulatory molecules and cytokine production of these cells. Both spontaneous and up-regulated costimulatory molecule expression and cytokine production were significantly suppressed by the addition of IL-10. Autologous T cell proliferation was stimulated by purified protein derivative (PPD) in IFN gamma-treated synovial cells and treatment of these synovial cells with IL-10 also suppressed T-cell proliferation. Our results suggest that IL-10 has an inhibitory effect on synovial cells and is an important immunoregulatory component of the cytokine network in rheumatoid arthritis. PMID- 9227326 TI - Polyclonal Th1 cells transfer oil-induced arthritis. AB - T-cells play a critical role in oil-induced arthritis (OIA) in DA rats. The present study focuses on the involvement of CD4/CD8 T cells in OIA by using adoptive transfer. Mitogen-activated T cells from DA rats previously injected with incomplete Freund's adjuvant (IFA) were depleted of CD4+ T cells or CD8+ T cells before transfer to irradiated naive receipients. The results indicate that CD4+ T cells are essential for the induction of passively induced OIA. However, in vitro blocking experiments with monoclonal antibodies (mAb) to the CD4 molecule of the T cells before transfer did not affect the passive OIA. Neither was passive OIA inhibited by treating the CD4+ T cells with mAb to intracellular adhesion molecule-1 (ICAM-1) in order to block cell-cell interactions or migration. The arthritogenic CD4+ T cells were sensitive, however, to in vitro treatment with mAb to the interleukin-2 receptor, which inhibited the disease or delayed the onset of passive OIA in recipients. The arthritogenic CD4+ T cells were also analysed for expression of specific T-cell receptor (TCR) variable (V) beta chains, critical for recognition of autoantigen, by utilizing V beta gene specific polymerase chain reaction (PCR). The results show a heterogeneous expression of V beta segments of the TCR, indicating a polyclonal origin of the pathogenic cells. Moreover, an investigation of the T helper (Th)1/Th2 status of the CD4+ T cells, defined by cytokine expression, was made at the mRNA level by using in situ hybridization. High numbers of interleukin-2 (IL-2) mRNA expressing cells and also interferon-gamma (IFN-gamma) and tumour necrosis factor-alpha (TNF alpha)-expressing cells could be identified. We conclude from this study that non immunogenic IFA triggers polyclonal, IL-2-dependent Th1 cells which induce arthritis. The contribution of the CD4 or ICAM-1 molecules for arthritis induction seem to be of minor importance. PMID- 9227327 TI - Neutrophils from the synovial fluid of patients with rheumatoid arthritis express the high affinity immunoglobulin G receptor, Fc gamma RI (CD64): role of immune complexes and cytokines in induction of receptor expression. AB - Neutrophils isolated from the synovial fluid of 16/24 patients with rheumatoid arthritis expressed Fc gamma RI (CD64), the high-affinity receptor for monomeric immunoglobulin G (IgG), on their cell surface. Receptor expression ranged from 17% to 168% of the level of expression obtained after incubation of control blood neutrophils with 100 U/ml interferon-gamma (IFN-gamma) for 24 hr in vitro. Similarly, mRNA for Fc gamma RI was detected in synovial fluid neutrophils from 12/15 patients and transcript levels ranged from 5% to 200% of the values obtained after treatment of blood neutrophils with IFN-gamma for 4 hr in vitro. No surface expression nor mRNA were detected in freshly isolated blood neutrophils from either patients or from healthy controls. Addition of cell-free synovial fluid to control blood neutrophils induced both mRNA and surface expression of Fc gamma RI to levels that were comparable to those achieved after addition of IFN-gamma. Neither soluble nor insoluble immune complexes appeared to be involved in induction of Fc gamma RI expression in spite of the ability of these complexes to induce protein biosynthesis. Synovial fluid-induced expression of Fc gamma RI was partially blocked by incubation with neutralizing IFN-gamma antibodies, whilst neutralizing interleukin (IL)-6 antibodies had little effect. Levels of IFN-gamma measured within these synovial fluids ranged from 0 to 2.7 U/ml, well within the range known to induce neutrophil Fc gamma RI expression. These data thus indicate that gene expression in synovial fluid neutrophils is selectively activated as the cells enter the diseased joint. Furthermore, these data indicate that induced expression of Fc gamma RI may alter the ability of infiltrating neutrophils to respond to IgG-containing immune complexes present in these joints. PMID- 9227328 TI - Activation of neutral sphingomyelinase in human neutrophils by polyunsaturated fatty acids. AB - Although unesterified polyunsaturated fatty acids (PUFA) have been shown to elicit marked changes in neutrophil function, the associated signal transduction processes require clarification. In this study we examined the effect of PUFA on the sphingomyelin (SM)-signalling cycle in human neutrophils. Treatment of neutrophils with eicosatetraenoic acid [arachidonic acid, 20:4(n-6)] caused a decrease in the mass of cellular SM and an increase in the level of ceramide. 20:4(n-6)-stimulated neutral sphingomyelinase (SMase) activity of the leucocytes in a time- and concentration-dependent manner. Other unsaturated fatty acids, docosahexaenoic [22:6(n-3)], eicosapentaenoic [20:5(n-3)], octadecenoic [oleic, 18:1(n-9)] and octadecadienoic [linoleic, 18:2(n-6)] acids also had the capacity to activate neutral SMase; however, certain 20:4(n-6) derivatives ?20:4(n-6) methyl ester [20:4(n-6)ME], 15-hydroperoxyeicosatetraenoic (15-HPETE) and 15 hydroxyeicosatetraenoic (15-HETE) acids?, very-long-chain PUFA ?tetracosatetraenoic [24:4(n-6)] and octacosatetraenoic [28:4(n-6)] acids? and saturated fatty acids [octadecanoic (stearic, 18:0) and eicosanoic (arachidic, 20:0) acids] had no significant effect. Activation of neutral SMase by 20:4(n-6) appeared to involve metabolism via 20:4(n-6)CoA (arachidonoyl CoA) and was not dependent on prostaglandin and leukotriene synthesis. All of the fatty acids and derivatives tested failed to activate acidic SMase of neutrophils. Ceramide was found to inhibit 20:4(n-6)-induced superoxide generation by the cells. It is envisaged that the PUFA-induced ceramide production in neutrophils plays a role in the regulation of biological responses. PMID- 9227329 TI - The involvement of macrophage-derived tumour necrosis factor and lipoxygenase products on the neutrophil recruitment induced by Clostridium difficile toxin B. AB - Clostridium difficile (Cd) toxins appear to mediate the inflammatory response in pseudomembranous colitis and/or colitis associated with the use of antibiotics. In contrast to Cd Toxin A (TxA), Cd Toxin B (TxB) has been reported not to promote fluid secretion or morphological damage in rabbits and hamsters and also does not induce neutrophil chemotaxis in vitro. However, TxB is about 1000 times more potent than TxA in stimulating the release of tumour necrosis factor-alpha (TNF-alpha) by cultured monocytes. In the present study, we investigated the ability of TxB to promote neutrophil migration into peritoneal cavities and subcutaneous air-pouches of rats. We also examined the role of resident peritoneal cells in this process as well as the inflammatory mediators involved. TxB caused a significant and dose-dependent neutrophil influx with a maximal response at 0.1 microgram/cavity after 4 hr. Depleting the peritoneal resident cell population by washing the peritoneal cavity or increasing this population by pretreating the animals with thioglycollate blocked and amplified the TxB-induced neutrophil migration, respectively. Pretreating the animals with MK886 (a lipoxygenase inhibitor), NDGA (a dual cyclo- and lipoxygenase inhibitor) or the glucocorticoid, dexamethasone, but not with indomethacin (a cyclo-oxygenase inhibitor), or BN52021 (a platelet-activating factor antagonist), inhibited the neutrophil migration evoked by TxB. Pretreatment with dexamethasone or the administration of anti-TNF-alpha serum into the air-pouches also significantly reduced the TxB-induced neutrophil migration. Supernatants from TxB-stimulated macrophages induced neutrophil migration when injected into the rat peritoneal cavity. This effect was attenuated by the addition of either MK886 or dexamethasone to the macrophage monolayer and by preincubating the supernatants with anti-TNF-alpha serum. TxB also stimulated the release of TNF-alpha by macrophages. Overall, these results suggest that TxB induces an intense neutrophil migration which is mediated by macrophage-derived TNF-alpha and lipoxygenase products. PMID- 9227330 TI - CR3-dependent phagocytosis by murine macrophages: different cytokines regulate ingestion of a defined CR3 ligand and complement-opsonized Cryptococcus neoformans. AB - Phagocytosis is a fundamental process in innate resistance to infection. We have used the pathogenic yeast Cryptococcus neoformans to study the interaction of this encapsulated organism with murine macrophages in vitro. In the absence of exogenous opsonins the encapsulated yeast is almost totally resistant to ingestion by murine macrophages. Owing to its ability to activate the alternative complement pathway, the anti-phagocytic properties of the polysaccharide capsule can be partially overcome following opsonization in vitro with non-immune mouse serum and subsequent phagocytosis via complement receptors. Here, we demonstrate the importance of the complement receptor type 3 (CR3) in in vitro phagocytosis of the yeast and in in vivo resistance to infection. In vitro, 70% of a population of resident murine macrophages are able to ingest C. neoformans and then only inefficiently (1-2 organisms per cell). Previously we have shown that tumour necrosis factor-alpha (TNF-alpha) and granulocyte-macrophage colony stimulating factor (GM-CSF) efficiently enhance ingestion of serum-opsonized encapsulated C. neoformans, and we now show that the cytokines convert a population of resident macrophages to a state where all the cells are competent for ingestion of large numbers of yeasts (6-8 per cell). We also show that these cytokines have a direct effect on CR3, as enhanced levels of complement-opsonized sheep red blood cells (EIgMC) bind to macrophages activated in this way. However, cytokines that have previously been shown to enhance phagocytosis of EIgMC have no effect on ingestion of encapsulated C. neoformans. These results demonstrate that the cytokines regulating CR3-dependent ingestion of C. neoformans are different to those regulating ingestion of EIgMC and reinforce the importance of studying pathogens rather than inert ligands in understanding the regulation of phagocytosis. PMID- 9227331 TI - Transcriptional activation of the interleukin-8 gene by platelet-activating factor in human peripheral blood monocytes. AB - Interleukin-8 (IL-8) is a member of the chemokine family and a potent neutrophil chemoattractant and activator. It is produced by a variety of cell types during inflammation. In the present work, we examined the regulation of IL-8 gene expression in monocytes by the pro-inflammatory lipid mediator, platelet activating factor (PAF). Stimulation of human peripheral blood monocytes with PAF augmented their release of IL-8. The enhancement of IL-8 secretion was associated with an increase in IL-8 mRNA expression. PAF induced a concentration- and time dependent augmentation of IL-8 mRNA accumulation. The response was maximal at PAF concentrations of 10-100 nM. The increased mRNA expression was evident after 1.5 hr of stimulation and persisted for 6 hr. Stimulation of monocytes with PAF, followed by arrest of de novo transcription with actinomycin D, indicated that PAF only marginally increased the stability of IL-8 mRNA. However, in vitro nuclear transcription demonstrated that the enhancement of IL-8 mRNA expression occurred mainly at the transcriptional level. The PAF-induced increase in IL-8 mRNA levels could be blocked with a PAF receptor antagonist. These results show, for the first time, that IL-8 gene expression and protein production can be upregulated by PAF. This interaction could be important in the development and amplification of the inflammatory response. PMID- 9227332 TI - Pattern of cytokine receptors expressed by human dendritic cells migrated from dermal explants. AB - Different reasons account for the lack of information about the expression of cytokine receptors on human dendritic cells (DC): (a) DC are a trace population; (b) the proteolytic treatment used to isolate DC may alter enzyme-sensitive epitopes; and (c) low numbers of receptors per cell. In the present work the expression of cytokine receptors was analysed by flow cytometry on the population of dermal DC (DDC) that spontaneously migrate from short-term culture dermal explants. DDC obtained after dermal culture were CD1alow, CD1b+, CD1c+, human leucocyte antigen (HLA)-DR+, CD11chigh, CD11b+ and CD32+. The DC lineage was confirmed by ultrastructural analysis. DDC expressed interleukin (IL)-1R type 1 (monoclonal antibody (mAb) hIL-1R1-M1; and 6B5); IL-1R type 2 (mAb hIL-1R2-M22); IL-2R alpha chain (mAb anti-Tac; and hIL-2R-M1) and IL-2R gamma chain (mAb 3B5; and AG14C). DDC did not stain for IL-2R beta chain using four mAbs recognizing two different epitopes of IL-2R beta (mAb 2R-B; Mik-beta 1; and CF1; Mik-beta 3, respectively). DDC were also positive for the cytokine binding chains (alpha chains) of IL-3R (mAb 9F5); IL-4R (mAb hIL-4R-M57; and S456C9); and IL-7R (mAb hIL-7R-M20; and R3434). DDC showed low levels of IL-6R alpha chain (mAb B-F19; B R6; and B-E23) and its signal transducer gp130 (mAb A2; and B1). DDC strongly expressed interferon-gamma receptor (IFN-gamma R) (mAb GIR-208) and were negative for IL-8R (mAb B-G20; and B-F25). All DDC were highly positive for granulocyte macrophage colony-stimulating factor receptor (GM-CSFR) alpha chain (mAb hGM-CSFR M1; SC06; SC04, and 8G6) and to a lesser extent for the common beta chain of GM CSFR, IL-3R and IL-5R (mAb 3D7). On the other hand, reactivity was not found for granulocyte colony-stimulating factor receptor (G-CSFR) (mAb hGCSFR-M1) nor macrophage colony-stimulating factor receptor (M-CSFR) (mAb 7-7A3-17) confirming the DC lineage of DDC. As previously reported for lymphoid DC, DDC expressed tumour necrosis factor receptort (TNFR) 75000 MW (mAb utr-1; hTNFR-M1; and MR2-1) but lacked TNFR 55000 MW (mAb htr-9; MR1-1; and MR1-2). In summary, DDC express receptors for a broad panel of cytokines, even receptors for cytokines whose effects on DC are still unknown (i.e. IL-2R alpha gamma; IL-6R alpha/gp 130; IL 7R alpha gamma). PMID- 9227333 TI - Immune enhancing effects of dehydroepiandrosterone and dehydroepiandrosterone sulphate and the role of steroid sulphatase. AB - Steroid hormones, such as glucocorticoids (GC), influence immune and inflammatory responses through their suppressive actions. Recent evidence suggests that another steroid hormone, dehydroepiandrosterone (DHEA), provides an immunostimulatory influence opposing the effect of GC. DHEA circulates in its inactive sulphated form, DHEAS, requiring conversion to DHEA by a steroid sulphatase (SS) enzyme for biological activity. Therefore, inhibition of SS activity may affect immune responses, allowing endogenous GC effects to predominate. We have shown that administration of DHEA and DHEAS in contact sensitization (CS) augments ear swelling by 39 and 46% respectively (P < 0.001). DHEAS at doses of 0.5, 5 and 50 mg/kg reverses the inhibitory effect of corticosterone (5 mg/kg) (P < 0.01). In CS, CT2251 (SS inhibitor) at 10 and 0.1 mg/kg inhibited ear swelling by 61 and 38% (P < 0.05) respectively. In addition, it inhibited DHEAS-augmented responses by 49 and 35% respectively (P < 0.05), with no effect on DHEA-augmented responses. DHEAS reversed CT2251 inhibition of the CS response with complete reversal at 50 mg/kg (P < 0.05). DHEAS and CT2251 appear to affect cellular infiltration into the ear, since DHEAS increased the number of lymphocytes by 63.8% and macrophages by 107% (P < 0.001), whereas CT2251 at 0.1 mg/kg decreased the number of lymphocytes by 65% (P < 0.001) and macrophages by 80% (P < 0.001). DHEAS, CT2251 and dexamethasone had no effect on oedema in the ear. From our data we have shown that steroid hormones, such as DHEA, have the potential to act as immunostimulatory factors in vivo. Inhibiting the conversion of DHEAS to DHEA by SS enzyme leads to an anti-inflammatory effect. PMID- 9227334 TI - Fractals in pathology. AB - Many natural objects, including most objects studied in pathology, have complex structural characteristics and the complexity of their structures, for example the degree of branching of vessels or the irregularity of a tumour boundary, remains at a constant level over a wide range of magnifications. These structures also have patterns that repeat themselves at different magnifications, a property known as scaling self-similarity. This has important implications for measurement of parameters such as length and area, since Euclidean measurements of these may be invalid. The fractal system of geometry overcomes the limitations of the Euclidean geometry for such objects and measurement of the fractal dimension gives an index of their space-filling properties. The fractal dimension may be measured using image analysis systems and the box-counting, divider (perimeter stepping) and pixel dilation methods have all been described in the published literature. Fractal analysis has found applications in the detection of coding of coding regions in DNA and measurement of the space-filling properties of tumours, blood vessels and neurones. Fractal concepts have also been usefully incorporated into models of biological processes, including epithelial cell growth, blood vessel growth, periodontal disease and viral infections. PMID- 9227335 TI - Neuroendocrine differentiation in lung cancer. AB - The human lung has a resident population of neuroendocrine (NE) cells. These are characterised by the presence of neurosecretory granules and by a number of histochemical and immunocytochemical reactions. NE differentiation is a defining feature of classical and atypical carcinoid tumours and is seen in most small cell lung cancers (SCLC). Such differentiation has also been described in up to one third of non-small cell lung cancers (NSCLC). As demonstrated in an accompanying paper in this issue of the Journal, the reporting of NE differentiation varies with the technical approach used and with the nature of the tissue specimen. This phenomenon is a reflection of the histological and biological heterogeneity of lung cancer and has not been shown to be of clinical utility. PMID- 9227336 TI - Making sense out of in situ PCR. AB - Given the limited sensitivity of existing in situ hybridization methods for detecting specific nucleic acid sequences, amplification in situ by means of the polymerase chain reaction (PCR) seems to be an attractive alternative. Recent studies using in situ PCR technology have not assessed the gain in signal strength that has been achieved, nor evaluated quantitatively the efficiency of amplification. An accompanying article in the current issue of the Journal examines the reproducibility and amplification efficiency of an RT-PCR in situ hybridization method that uses a sense probe, capable of detecting only amplified target sequences. The amplification procedure resulted in approximately 3-6-fold increased sensitivity that depended upon cell type and disease status. PMID- 9227337 TI - A comparison of NSP-reticulons with conventional neuroendocrine markers in immunophenotyping of lung cancers. AB - Neuroendocrine-specific protein (NSP)-reticulons are endoplasmic reticulum associated protein complexes, which have been identified as markers for neuroendocrine differentiation. In this study, the expression of two members of the family of NSP-reticulons, NSP-A and NSP-C, have been investigated in different types of lung cancer and compared with the expression patterns of five conventional neuroendocrine markers, the neural cell adhesion molecule (NCAM), synaptophysin, chromogranin A, Leu-7, and neurofilament proteins. NSP-A and NSP-C antibodies were reactive with most carcinoid tumour and small cell lung carcinoma (SCLC) cases, while atypical carcinoid tumours showed a variable expression. In the total group of neuroendocrine tumours, a high concordance of expression was found between NSP-A and NSP-C, while their expression correlated well with NCAM and synaptophysin positivity. Chromogranin A, Leu-7, and neurofilament proteins were shown to be expressed to a limited extent in these neuroendocrine tumours. In a selected group of non-SCLCs known to exhibit neuroendocrine features, NSP-A expression was detected at much higher frequency than NSP-C. In virtually all NSP A positive cases, this expression was associated with one or more of the other neuroendocrine markers. NSP-A expression showed a stronger correlation with conventional neuroendocrine markers than NCAM. In detecting neuroendocrine differentiation in non-SCLC, NSP-A is more sensitive than synaptophysin, chromogranin A, Leu-7, and neurofilament proteins. It is concluded that NSP reticulons are valuable markers in the diagnosis of neuroendocrine differentiation in non-SCLC and should be used in conjunction with NCAM. PMID- 9227338 TI - Quantification of vitamin D receptor mRNA in tissue sections demonstrates the relative limitations of in situ-reverse transcriptase-polymerase chain reaction. AB - In situ-reverse transcriptase-polymerase chain reaction (IS-RT-PCR) is a recently described technique that is used to localize low levels of mRNA within cells and tissue sections. One of the major criticisms levelled at this technique is that positive results may be meaningless, as amplification is required to demonstrate the transcripts of interest. The use of IS-RT-PCR to demonstrate mRNA for receptors for 1,25-dihydroxyvitamin D3 (VDR) in sections of human kidney and bone has previously been described. To ascertain whether the levels of VDR mRNA detected following IS-RT-PCR were transcriptionally significant, computerized image analysis was used to determine the mean silver grain density in human kidney and bone cells following conventional in situ hybridization and after various cycles of IS-RT-PCR. Only a few cycles of PCR were needed to produce an optimum signal, but amplification of signal following IS-RT-PCR was found to be relatively inefficient. Following the optimum number of cycles of IS-RT-PCR in kidney sections, there was a less than four-fold increase in signal. Similarly, in bone, the optimum signal detected was only approximately five times greater than that found with conventional in situ hybridization. These results clearly demonstrate that the increase in signal following IS-RT-PCR follows a more linear pattern and is relatively inefficient, compared with the usual exponential increase with conventional solution phase RT-PCR. PMID- 9227339 TI - The pathological and biological nature of screen-detected breast carcinomas: a morphological and immunohistochemical study. AB - Traditional and immunohistochemical markers of prognosis were examined in 455 mammary carcinomas derived from breast cancer screening and compared with those of 277 carcinomas presenting symptomatically over the same period. Tumours detected by population screening under the U.K. National Health Service Programme do not differ from those detected by other screening projects, but compared with symptomatic cancers, screen-detected cancers are more likely to be in situ and if invasive, to be smaller, of lower grade, and to have invaded vessels, perineural spaces, and lymph nodes less frequently. Tubular and cribriform types are more often represented in screened patients. Immunohistochemical markers which have been proposed as being related to likely tumour behaviour (epidermal growth factor receptor, c-erbB-2 protein, oestrogen and progesterone receptors, cathepsin D, p53, and retinoblastoma protein) do not distinguish screen-detected from 'clinical' cancers. It is concluded that cancers diagnosed at screening do not differ biologically from those presenting clinically, but are the same lesions detected at an earlier stage of their natural history. PMID- 9227340 TI - The spectrum of 67-kD laminin receptor expression in breast carcinoma progression. AB - Laminin is a glycoprotein of the basement membrane (BM), involved in a variety of normal and pathological cellular events including tumour invasion and metastasis. Cells bind laminin through different types of receptor. The 67-kD laminin receptor (67LR) is a cell-surface protein which binds laminin with high affinity. 67LR expression has been shown to increase in neoplastic cells, compared with normal tissues, and 67LR seems to play an important role during the first steps of neoplastic progression. In this study, 67LR expression was analysed during the morphological phases of breast cancer progression from normal tissue to invasive carcinoma. A total of 506 formalin-fixed, paraffin-embedded normal breast structures and lesions were stained by immunohistochemistry usign the MLuC5 monoclonal antibody, which is specific for 67LR. The results show that in normal breast and in any kind of breast lesion, myoepithelial and endothelial cells express 67LR. While 67LR is not seen in the epithelium of normal breast, cysts, adenosis, and benign tumours, it is expressed in the epithelial cells of several hyperplasias and carcinomas in situ, both ductal and lobular, as well as in all invasive carcinomas. The 67LR-positive cell subpopulation expands from hyperplastic lesions to invasive carcinoma, suggesting that 67LR could be related to the induction and progression of breast cancer. PMID- 9227341 TI - 3-D histo-radiographic comparison of surgical clearances of microcalcified lesions in breast localization biopsies. AB - There is controversy as to the value of the radiological or pathological estimation of surgical clearance of microcalcifying breast lesions. An important part of this issue has been addressed by coordinated three-dimensional radiographic and histological examination of a prospective consecutive series of 40 benign and malignant mammographically detected lesions in surgical breast biopsy specimens containing microcalcifications, including 20 cases of ductal carcinoma in situ. They were radiographed from four viewpoints by means of rotation in a radiolucent tetrahedral container. The planes of histological examination were then chosen to correspond to the radiographic view showing the minimum separation of the edge of the specimen and the outermost microcalcification. There was a close correlation (Spearman ranked) between the least tetrahedral radiographic distance and the corresponding histological distance separating the surgical margin of excision. There were, however, incompatible Wilcoxon signed ranking orders when comparing the least tetrahedral distance or the histological distance with all four single radiographic views, including the conventional specimen radiographic view. Two-dimensional specimen mammography and standardized histological examination are suboptimal and may thus have contributed to confusion as to the value of determining adequate surgical excision of ductal carcinoma in situ of the breast. Although labour-intensive, use of four-view radiography and choice of the appropriate plane of histological examination give a better correlation of the radiographic estimates of surgical clearance with histology than single-view specimen radiography and arbitrary histological sectioning. PMID- 9227342 TI - Point mutations and nucleotide insertions in the MDM2 zinc finger structure of human tumours. AB - This study investigates the human oncoprotein MDM2, which interferes with regulation of cell division and apoptosis. Fifteen mixed-type follicular non Hodgkin's lymphomas, ten leukaemias, two hepatocellular carcinomas, one osteosarcoma, and ten normal cell lines (fibroblasts, osteoblasts, mesothelium, peripheral lymphocytes) were tested for MDM2 expression and MDM2 gene mutation by reverse transcriptase-polymerase chain reaction (RT-PCR), immunocytochemistry, and nucleotide sequence analysis. Two follicular lymphomas, three leukaemias, both hepatocellular carcinomas, and the osteosarcoma sample showed transcription of the activated MDM2 gene. These samples lacked amplified MDM2 genes and carried mis-sense, non-sense and frame-shift mutations in a zinc finger region of MDM2, altering the amino acid sequence or causing premature termination of transcription. The mis-sense mutations were found in tumour cells that showed significant accumulation of MDM2 and lack of nuclear p53. Non-sense mutations and frame-shift mutations were found in tumours lacking MDM2 proteins. The mutations may affect the biological properties of MDM2 proteins. PMID- 9227343 TI - Ki67 labelling index in gastric carcinomas. An immunohistochemical study using double staining for the evaluation of the proliferative activity of diffuse-type carcinomas. AB - The aim of the present study was to clarify the conflicting recorded data on the proliferative features of gastric carcinoma. The Ki67 labelling index (Ki67 LI) was evaluated using MIB-1 in 43 carcinomas (24 diffuse and 19 intestinal). In 18 cases, differential counts were performed in superficial and deep layers. In ten diffuse carcinomas with a prominent desmoplastic response, Ki67 LI was evaluated in sections double-stained with MIB-1 and CAM5.2. Flow cytometry was performed in 26 cases. Ki67 LI of diffuse carcinomas (36.3 +/- 19.0) was not significantly different from that of intestinal carcinomas (28.2 +/- 18.5). Ki67 LI was significantly higher (P = 0.006) in superficial than in deep areas (41.9 +/- 22.7 and 29.7 +/- 19.7, respectively) regardless of histological tumour type. No significant relationship was observed between Ki67 LI and wall invasion, lymph node metastasis, vascular invasion or ploidy. The following conclusions were drawn: double immunostaining techniques are apparently the best way to overcome the underestimation of cell proliferation in diffuse gastric carcinomas with a prominent desmoplastic response; the diffuse and intestinal types of gastric carcinoma have proliferation rates within the same range, even when the comparison is restricted to diploid tumours; and, finally, the major pool of proliferating cells resides in the superficial areas of gastric carcinomas, regardless of the histotype, which should be taken into consideration when overall counts are performed, using either immunohistochemical markers in tissue sections or suspensions of nuclei in flow cytometry. PMID- 9227344 TI - Automated location of dysplastic fields in colorectal histology using image texture analysis. AB - Automation in histopathology is an attractive concept and recent advances in the application of computerized expert systems and machine vision have made automated image analysis of histological images possible. Systems capable of complete automation not only require the ability to segment tissue features and grade histological abnormalities, but, must also be capable of locating diagnostically useful areas from within complex histological scenes. This is the first stage of the diagnostic process. The object of this study was to develop criteria for the automatic identification of focal areas of colorectal dysplasia from a background of histologically normal tissue. Fields of view representing normal colorectal mucosa (n = 120) and dysplastic mucosa (n = 120) were digitally captured and subjected to image texture analysis. Two features were selected as being the most important in the discrimination of normal and adenomatous colorectal mucosa. The first was a feature of the co-occurrence matrix and the second was the number of low optical density pixels in the image. A linear classification rule defined using these two features was capable of correctly classifying 86 per cent of a series of training images into their correct groups. In addition, large histological scenes were digitally captured, split into their component images, analysed according to texture, and classified as normal or abnormal using the previously defined classification rule. Maps of the histological scenes were constructed and in most cases, dysplastic colorectal mucosa was correctly identified on the basis of image texture: 83 per cent of test images were correctly classified. This study demonstrates that abnormalities in low-power tissue morphology can be identified using quantitative image analysis. The identification of diagnostically useful fields advances the potential of automated systems in histopathology: these regions could than be scrutinized at high power using knowledge-guided image segmentation for disease grading. Systems of this kind have the potential to provide objectivity, unbiased sampling, and valuable diagnostic decision support. PMID- 9227345 TI - Interleukin 6 gene-transfected mouse mammary adenocarcinoma: tumour cell growth and metastatic potential. AB - Cells from the spontaneous metastatic TSA mammary adenocarcinoma of BALB/C mouse were transfected with the murine (interleukin-6) IL6 gene. The clone (TSA-IL6) secreting the largest amount of IL6 displayed an in vitro increased growth rate compared with that of TSA cells transfected with the neomycin resistance gene only (TSA-neo). TSA-IL6 cell colonies consisted mainly of fusiform cells and TSA neo colonies of polygonal cells. When subcutaneously (s.c.) injected in syngeneic mice, TSA-IL6 cells gave rise to tumours that grew significantly slower than TSA neo cell tumours. Microscopically, TSA-IL6 tumours displayed a fascicular pattern of growth, associated with a very scanty macrophage infiltrate. S.c. TSA-IL6 tumours were significantly less metastatic than TSA-neo tumours. By contrast, following intravenous (i.v.) challenge, TSA-IL6 cells produced 5-7 times more lung metastases than TSA-neo cells. The i.v. TSA-IL6 cell lung metastases showed a marked macrophage infiltrate and a rich vascularization. The high in vitro TSA IL6 cell growth rate is attributable to the IL6-induced production of growth factors, some of which possess heparin-binding properties, such as amphiregulin. The differences in vascularization and macrophage infiltrate may underlie the observed differences between s.c. TSA-IL6 tumour growth with low spontaneous metastatic potential and the widespread growth of i.v. metastasis. PMID- 9227346 TI - Terminal deoxynucleotidyl transferase staining of malignant lymphomas in paraffin sections: a useful method for the diagnosis of lymphoblastic lymphoma. AB - Terminal deoxynucleotidyl transferase (TdT) is a DNA polymerase located in the cell nucleus which catalyses the polymerization of deoxynucleotides at the 3'hydroxyl ends of oligo- or polydeoxynucleotide initiators without a template. TdT is known as a useful marker for the diagnosis of acute lymphoblastic leukaemia/lymphoma, but its detection usually requires fresh tissue specimens or cell suspensions, using either an enzyme analysis or immuno-fluorescence or peroxidase staining. Until the recent development of the use of microwave-treated paraffin sections for immunoperoxidase staining, detection of TdT in paraffin sections required rather complicated processes. This new simple technique was applied to paraffin sections from the tumour tissue specimens of 16 patients with lymphoblastic lymphoma and of seven patients with non-endemic Burkitt's lymphoma, which is sometimes difficult to differentiate from lymphoblastic lymphoma because of their similar clinicopathological characteristics. In addition, as a control, ten cases each were examined of adult T-cell leukaemia/lymphoma (ATLL) and angioimmunoblastic lymphoma (AILD), which are both peripheral T-cell lymphomas. The tumour cells from 15 of the 16 (94 per cent) patients with lymphoblastic lymphoma were found to be TdT-positive. The specificity of the anti-TdT antibody used was confirmed by immunoblot and the specific 60 kD band was detected only in a specimen of lymphoblastic lymphoma. These results show that the immunostaining of TdT on paraffin-embedded sections is a useful method for differentiating lymphoblastic lymphoma from other lymphomas. This method is applicable to a routine diagnostic service. PMID- 9227347 TI - Osteoclasts are capable of particle phagocytosis and bone resorption. AB - Osteoclasts are multinucleated cells specialized for the function of lacunar bone resorption. Although they are known to be capable of phagocytosis of inert particles, it is not known whether this abolishes their ability to respond to hormones or to form resorption lacunae. Human and rat osteoclasts were isolated from giant cell tumours of bone and rat long bones, respectively, and cultured on coverslips and cortical bone slices, both in the presence and in the absence of particles of latex (1 micron diameter) and polymethylmethacrylate (PMMA) (< 50 microns). By light microscopy, it was evident that osteoclasts which had phagocytosed both latex and PMMA particles remained responsive to calcitonin. Osteoclast phagocytosis of particles was also evident on scanning electron microscopy, where it could also be seen that these cells were associated with the formation of resorption lacunae. These findings underline the fact that the osteoclast is a true member of the mononuclear phagocyte system and that phagocytosis does not abrogate either its hormonal response to calcitonin or its highly specialized function of bone resorption. That osteoclasts which have phagocytosed biomaterial particles such as PMMA are still able to carry out lacunar bone resorption is of interest in clinical conditions such as aseptic loosening, where a heavy foreign body particle load is often associated with extensive bone resorption. PMID- 9227348 TI - Properties of multinucleated giant cells in a new in vitro model for human granuloma formation. AB - Multinucleated giant cells (MGCs) are a key feature of granulomas. They have been studied with respect to the mechanism and regulation of their formation, but the function of these cells still remains elusive. A new method for the in vitro generation of granulomas was developed and characterized in which L3 larvae of Nippostrongylus brasiliensis, as a target for the cellular response, were co incubated with human mononuclear blood cells. The development of epithelioid cells and MGCs was observed and single isolated MGCs were analysed by the reverse transcriptase polymerase chain reaction method. The presence of tumour necrosis factor alpha (TNF alpha), interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS) transcripts in MGCs was demonstrated. It is proposed that MGCs in the granuloma model may in part represent an active cellular constituent involved in granuloma formation and turnover and in the destruction of the irritant. PMID- 9227349 TI - The role of monocyte chemoattractant protein-1 (MCP-1) in the pathogenesis of collagen-induced arthritis in rats. AB - Collagen-induced arthritis was produced in rats by intradermal immunization with type II collagen and the expression and production of monocyte chemoattractant protein-1 (MCP-1) were examined by immunohistochemistry, enzyme-linked immunosorbent assay (ELISA), and Northern blot analysis. Two to three weeks after the immunization, the hindfeet showed swelling and redness, followed by the development of severe arthritis, particularly in the ankle joints. During this period, prominent infiltration of neutrophils and macrophages was observed. Sandwich ELISA and Northern blot analysis revealed that MCP-1 concentrations in the joint lavages and MCP-1 mRNA levels in the joint tissues both peaked at 2 weeks after the immunization. By immunohistochemistry, various types of cells, particularly neutrophils, macrophages, synovial cells, and vascular endothelial cells, stained positively for MCP-1. Finally, injection of a neutralizing monoclonal antibody against rat MCP-1 significantly decreased the number of exudate macrophages in the lesions and reduced the ankle swelling by about 30 per cent compared with controls. These results suggest that MCP-1 plays a critical role in this model in the recruitment of monocytes and in the development of arthritis. PMID- 9227350 TI - Mast cell distribution, activation, and phenotype in atherosclerotic lesions of human carotid arteries. AB - Immunohistochemical staining for mast cell tryptase and chymase was used to examine the distribution, activation, and tryptase/chymase phenotype of mast cells (MCs) in 250 samples of atherosclerotic lesions (type I to VI) of human carotid arteries. Dual immunolocalization and histochemical techniques were used to identify the associations of MCs with macrophages, smooth muscle cells, and extracellular matrix components. Whereas normal carotid arteries contained very few MCs within the intima, atherosclerotic lesions showed increased MC numbers with variable focal accumulations. MCs were identifiable from the earliest stages of atherosclerosis, and especially at the shoulder regions of the fully formed atheroma. They were observed in close association with macrophages (HAM56 positive) and extracellular lipid, as well as at sites of foam cell formation. MCs and diffuse tryptase staining were also evident within sites of new calcification and around small calcified deposits. Extensive MC activation/degranulation, as judged by diffuse extracellular tryptase staining, was a common feature of the advanced atherosclerotic plaques complicated by fissure, haemorrhage, and thrombus formation. Moreover, such sites of extracellular MC tryptase were often associated with localized oedema and disruption of the stromal matrix. MCs which contained both tryptase and chymase (the MCTC phenotype) represented approximately 80-95 per cent of all MCs. These studies are the first to demonstrate significant numbers and focal accumulations of MCs in all developmental stages of atherosclerotic carotid arteries. Since MCs contain or express a variety of potent mediators, their release could profoundly influence the development and pathological complications of atherosclerotic plaques. PMID- 9227351 TI - Substrate utilization and maturation of cumulus cell-enclosed mouse oocytes: evidence that pyruvate oxidation does not mediate meiotic induction. AB - This study was performed to address the possible role of pyruvate in meiotic induction in mouse oocytes. Cumulus cell-enclosed oocytes from primed, immature mice were cultured in 7.5 microliters microdrops under oil for 9 or 18 h in medium containing 4 mmol hypoxanthine l-1 plus 0.23 mmol pyruvate l-1, 1 mmol pyruvate l-1, or 1 mmol pyruvate l-1 plus 5.5 mmol glucose l-1. When compared with cultures containing 0.23 mmol pyruvate l-1, 1 mmol pyruvate l-1 induced germinal vesicle breakdown, and this was preceded by an increase in pyruvate utilization. Addition of glucose prevented both the increase in pyruvate consumption and the meiotic induction. When different combinations of pyruvate and glucose were tested on oocyte maturation in microdrop cultures, a high concentration of pyruvate or glucose alone was stimulatory to maturation. Addition of the complementary energy substrate prevented the induction of germinal vesicle breakdown and reduced the amount of substrate consumption. During spontaneous maturation in vitro, oocyte-cumulus cell complexes consumed glucose for the first 3 h; however, during the second 3 h period, which followed germinal vesicle breakdown, glucose consumption decreased and net pyruvate utilization was initiated. Treatment of hypoxanthine-arrested oocytes with dichloroacetate, an activator of pyruvate dehydrogenase, stimulated pyruvate consumption but had no effect on germinal vesicle breakdown. Although FSH stimulates meiotic resumption, no changes in pyruvate consumption were observed in response to this gonadotrophin. Measurement of oxygen consumption by hypoxanthine-treated complexes revealed no effect of high concentrations of pyruvate on respiration, and FSH treatment resulted in a suppression of oxygen utilization. These data indicate that, in mouse oocyte-cumulus cell complexes, pyruvate and glucose can each modulate metabolism of the other substrate, and the can significantly influence meiotic maturation of the oocyte. In addition, augmentation of pyruvate oxidation does not appear to play a mediating role in meiotic induction triggered by energy substrate manipulation or gonadotrophin treatment. PMID- 9227352 TI - Effect of cryoprotectants on the survival of follicles in frozen mouse ovaries. AB - Ovaries from 10-day-old mice were exposed to 1.5 mol l-1 dimethylsulfoxide, 1,2 propanediol, ethanediol or glycerol for 5-60 min at room temperature before freezing. Follicles in fresh and frozen ovaries were counted and scored as normal or damaged in stained serial sections. More primordial follicles survived in ovaries frozen in dimethylsulfoxide, 1,2-propanediol and ethanediol (81-94%) than in those frozen in glycerol (4-28%). Prolonged exposure to ethanediol (60 min) before cooling decreased the survival rate, while increasing the exposure to glycerol (> or = 12 min) increased the survival rate. Fewer than 49% of primary follicles survived freezing. After transfer underneath the kidney capsules of ovariectomized immunodeficient recipients, there was no difference in the establishment of grafts of fresh (92%) and frozen (90%) ovaries, the number of recipients showing vaginal cornification (fresh, 91%, frozen 96%) or the latency of cornification (11 days). Fifteen days after transplantation, similar numbers of follicles remained in grafts of fresh ovaries, in ovaries frozen in dimethylsulfoxide and 1,2-propanediol, and in ovaries frozen after exposure to ethanediol for 5-30 min. Overall, the total number of follicles remaining in grafts of ovaries frozen in dimethylsulfoxide and 1,2-propanediol represented 42 46% of follicles present in ungrafted ovaries. This was not significantly different from grafts of fresh ovaries (63%). Dimethylsulfoxide and 1,2 propanediol are the most effective cryoprotectants for 10-day-old mouse ovaries. The majority of follicles are lost during graft establishment. PMID- 9227353 TI - Synchronization of cell division in eight-cell bovine embryos produced in vitro: effects of 6-dimethylaminopurine. AB - The methods used to achieve blastomere cell cycle synchronization in embryos used as nuclear donors during embryo reconstruction have been largely unsuccessful. The aim of this study was to determine the reliability of 6-dimethylaminopurine (6-DMAP), an inhibitor of maturation promoting factor, to half and to synchronize blastomere division in cleavage stage bovine embryos. A second goal was to assess its reversibility and toxicity in vitro. Eight-cell stage embryos obtained at 58 h after insemination were treated with several concentrations of 6-DMAP for 12 h. Treated embryos were assessed for cleavage arrest, chromatin morphology, DNA synthesis, histone H1 and scored for blastocyst formation and for hatching rate. They were subsequently fixed and the number of nuclei counted. Complete arrest of cell division was observed at concentrations of 3 mmol 6-DMAP l-1 and above. At these concentrations, interphase nuclei in arrest were noticeably larger compared with interphase nuclei of eight-cell control embryos. Removal from 6-DMAP led to release from cleavage arrest and was followed by synchronized mitosis, histone H1 kinase deactivation and re-entry into interphase within 4-5 h. Twenty-nine per cent of interphase nuclei were synthesizing DNA at the end of the 12 h treatment as indicated by BrdU analysis. At 2 h after removal from 6-DMAP, an abrupt decrease to 9% BrdU-positive nuclei was observed followed by an increase to 39% by 6 h and a decrease to 28% at 10 h. The ability of treated embryos to reach the blastocyst stage in vitro and the number of cells per blastocyst were reduced. These results indicate that 6-DMAP can reversibly arrest and synchronize cleavage to the fifth cell cycle in eight-cell bovine embryos. Although a decrease was observed in the proportion of blastocysts obtained after treatment, it is concluded that 6-DMAP is a useful tool for synchronization studies requiring donor nuclei at metaphase before fusion to recipient oocyte. PMID- 9227354 TI - Sociosexual behaviour and paternity in procarbazine-exposed rats with or without regional testicular circulatory isolation. AB - Male Sprague-Dawley rats were used in two experiments in which a procarbazine bolus (400 mg kg-1 body mass) was administered with or without testicular circulatory isolation in the form of brief clamping of the spermatic cord and gubernaculum during drug administration. Separate tests of aggressiveness, sexual motivation, copulatory performance and paternity over the subsequent 6 weeks were used to assess functional changes resulting from testicular circulatory isolation. Experiment 1 compared intermale aggression and sexual motivation of animals in groups receiving procarbazine plus testicular circulatory isolation lasting 0, 15 or 45 min with that of animals in control groups with no clamp and no drug. Experiment 2 used a 2 x 2 factorial design to evaluate sexual performance and resulting paternity in animals 2 months after testicular circulatory isolation and drug exposure compared with that in control animals. Procarbazine treatment induced minimal disruption of normal interest in a receptive female, copulatory measures (intromissions or ejaculations) and structural integrity of seminal vesicles, bulbospongiosus muscles and ventral prostate glands. Animals exposed to the drug without testicular circulatory isolation were significantly less aggressive than animals in other groups. The most profound influence of procarbazine was on paternity. Males exposed to procarbazine with or without testicular circulatory isolation impregnated notably fewer females than did control males that were not exposed to the drug. There was no evidence of recovery of normal fertility up to 10 weeks after exposure to the drug. In conclusion, the deleterious influence of procarbazine on androgen sensitive processes appears to be specific to intermale aggression and to fertility. The testicular circulatory isolation technique, for 45 min in particular, softened the impact of the drug on social behaviour, although procarbazine suppressed fecundity even with testicular circulatory isolation. PMID- 9227355 TI - Male pronuclear formation and early embryonic development of hamster oocytes matured in vitro with gonadotrophins, amino acids and cysteamine. AB - Male pronuclear (MPN) formation in oocytes after in vitro maturation (IVM) was compared with that of matured follicular oocytes that had matured in vivo (controls). Cumulus-oocyte complexes were matured in vitro for 13 h in modified Tyrode's solution (TLP-PVA); cumulus-free oocytes were then incubated in 20% oviductal fluid for 3 h, and washed and capacitated spermatozoa were added. MPN formation was significantly lower (P < 0.05) in IVM oocytes 3 to 12 h after insemination (0 to 34%, respectively) than in control oocytes (range, 98-100%). Female pronuclear formation was 84-100% in controls and IVM oocytes, but spermatozoa incompletely decondensed in IVM oocytes. The addition of 10 mumol l-1 during IVM, significantly increased (P < 0.05) MPN formation (from 17% in the absence of cysteine to 47% in the presence of cysteine), but was lower than that in controls (88%). During IVM, the addition of 10% serum or gonadotrophins (FSH and LH) with or without amino acids did not support MPN formation without cysteamine, whereas the treatment with gonadotrophins and 11 amino acids plus 200 mumol cysteamine l-1 (82%) equalled controls (92%). Development of oocytes after IVM (in 0, 10, 20% serum) in TLP-PVA, gonadotrophins, 11 amino acids and 200 mumol cysteamine l-1 was compared with development in controls. Of the IVM treatments, 20% serum was inferior at fertilization, but yielded the highest percentage of fertilized oocytes developing to or beyond the four-cell stage (20% serum versus controls, respectively); fertilized oocytes, 75% versus 88%; > or = four-cell embryo, 40% versus 53%; blastocyst, 8% versus 14%. It was concluded that during IVM, gonadotrophins plus 11 amino acids interacted with cysteamine, enhancing the decondensation of spermatozoa and MPN formation; oocytes matured in this medium with 20% serum were fertilized and some developed to the blastocyst stage. PMID- 9227356 TI - The relationship between the dietary provision of alpha-tocopherol and the concentration of this vitamin in the semen of chicken: effects on lipid composition and susceptibility to peroxidation. AB - This study is an attempt to enhance the resistance of chicken semen to peroxidative damage by supplementing the diet of cockerels with the major lipid soluble antioxidant alpha-tocopherol. Cockerels at 6 months of age were fed for 8 weeks with feed containing 0, 20, 200 or 1000 mg alpha-tocopherol kg-1. Semen was collected during the final 2 weeks of the supplementation period and the concentrations of alpha-tocopherol in the spermatozoa and the seminal plasma were determined. The concentrations of alpha-tocopherol in whole semen, spermatozoa and seminal plasma were approximately twice as high when the supplementation was 200 mg kg-1 compared with when supplementation was 20 mg kg-1; however, supplementation at 1000 mg kg-1 did not achieve any further increase in these concentrations of alpha-tocopherol. Thus, the concentration of alpha-tocopherol in semen displays only a limited responsiveness to manipulation by dietary means. In contrast, the concentrations of the vitamin in the testes and liver were found to be much more amenable to dietary manipulation, exhibiting increases of six seven-fold over the whole range of supplementation. However, the dietary-induced increase in the alpha-tocopherol content of semen did result in a significant reduction in the susceptibility of the semen to lipid peroxidation. A further effect of enhancing the concentration of this vitamin in the semen was a significant increase in the proportions of C20-22 polyunsaturated fatty acids in the sperm phospholipids. In addition, the proportion of phosphatidylethanolamine in the phospholipid was increased whereas that of sphingomyelin was reduced at the higher concentrations of alpha-tocopherol supplementation. Thus, an increased dietary intake of alpha-tocopherol does produce beneficial changes in the antioxidant capacity and lipid profile of semen, albeit to a relatively limited extent. PMID- 9227357 TI - Effects of dietary supplementation with alpha-linolenic acid on the phospholipid fatty acid composition and quality of spermatozoa in cockerel from 24 to 72 weeks of age. AB - The aim of this study was to investigate the effects of supplementing the diet of the male chicken with alpha-linolenic acid (18:3n-3) on the phospholipid fatty acid composition, motility and fertilizing ability of chicken spermatozoa. The birds in the control group received a diet supplemented with soybean oil rich in linoleic acid (18:2n-6) whereas those in the test group were supplemented with linseed oil rich in alpha-linolenic acid. A number of age-related changes in the lipid parameters of the spermatozoa were observed in control birds. Between 24 and 72 weeks of age the amount of total lipid in the spermatozoa of control birds increased by approximately 2.4 times and the proportions of cholesterol and free fatty acid also increased significantly, whereas the proportions of phospholipid and triacylglycerol decreased. In addition, the proportion of phosphatidylcholine in the total phospholipid increased, whereas the proportion of phosphatidylserine decreased during the same period. The proportion of docosatetraenoic acid (22:4n 6) in the phospholipid decreased significantly between 24 and 72 weeks of age. The concentration of spermatozoa in the semen of control birds increased to a maximum at week 39 and had decreased significantly by week 72. Supplementation with alpha-linolenic acid had little or no effect on the proportion of docosahexaenoic acid (22:6n-3) in the phospholipid profile of the spermatozoa. However, supplementation with alpha-linolenic acid did produce a significant but small increase in the proportion of docosapentaenoic acid (22:5n-3) recorded at 39 and 54 weeks. Thus, this study shows that the fatty acid composition of the sperm phospholipid demonstrates a marked resistance to dietary manipulation. Supplementation with alpha-linolenic acid significantly enhanced semen fertility at week 39. The results suggest that the small increase in the proportion of n-3 fatty acids in the sperm phospholipids induced by enriching the diet with alpha linolenic acid is associated with a significant improvement in semen quality at 39 weeks of age. PMID- 9227358 TI - Apoptosis pattern elicited by oestradiol treatment of the seminiferous epithelium of the adult rat. AB - It is widely assumed that oestrogen administration in the male mimics hypophysectomy by suppressing gonadotrophin secretion. Nevertheless, oestradiol treatment can increase germ-cell apoptosis mainly at stages IV-X of the spermatogenic cycle, rather than at stage VII when apoptotic germ-cell death is mainly triggered by gonadotrophin withdrawal caused by hypophysectomy. Since the roles of testicular oestrogens in spermatogenic regulation, if any, are unknown, we re-evaluated the germ-cell types that undergo apoptosis after oestradiol treatment. Adult male rats were injected daily with 50 micrograms oestradiol, oestradiol plus testosterone propionate (25 mg every 3 days) or oestradiol plus human menopausal gonadotrophin (equivalent to 25 iu FSH plus 25 iu LH) for 15 days. Apoptosis was assessed by in situ 3'-end labelling of internucleosomal DNA fragments in plastic semithin sections; the germ-cell types involved were identified by high-resolution light microscopy. The quantitative analysis of our results shows that the apoptosis pattern elicited by oestradiol treatment of the seminiferous epithelium differs from that reported to be caused by gonadotrophin or testosterone withdrawal, suggesting a possible role for oestradiol in the modulation of germ-cell death in the adult testis of the rat. PMID- 9227359 TI - Investigation of the association between the presence of cytoplasmic residues on the human sperm midpiece and defective sperm function. AB - Defective sperm function has been identified as one of the most common causes of human infertility. The aim of this investigation was to identify whether the presence of retained cytoplasm on the human sperm midpiece is associated with defective sperm function. Statistical analysis of data demonstrated a strong negative correlation between the presence of residual cytoplasm on the midpiece of spermatozoa in the inseminate and fertilization rate during IVF. Significant negative correlations were also identified between the percentage of spermatozoa in the ejaculate bearing cytoplasmic residues and (i) spermatozoa having membrane integrity and (ii) sperm concentration. A highly significant positive correlation was also revealed between the percentage of spermatozoa in the ejaculate with membrane integrity and the percentage of motile spermatozoa. These correlations suggest that retained cytoplasm is a cause of subfertility. Measurements of the percentage of spermatozoa bearing residual cytoplasm in the IVF inseminate could provide the basis for a simple predictive test before IVF. PMID- 9227360 TI - Motility of spermatozoa in hydrosalpingeal and follicular fluid of pigs. AB - Hydrosalpinges were created to collect adequate volumes of fluid during pre-, peri- and postovulatory intervals from the ampulla, ampullary-isthmic junction and the isthmic-utero-tubal junction of the oviducts from Large White gilts that had exhibited at least two natural oestrous cycles. The accumulated fluids, follicular fluid and Butschwiler's medium were compared for their effects on various parameters of boar sperm motility using the CellSoft, computer-assisted, digital image analysis system. Sperm velocity (micron s-1 +/- SEM) was significantly higher (P < 0.05) in follicular fluid (84 +/- 3; n = 5) than in fluids from the ampulla during peri- and early postovulatory intervals, and from the isthmic-utero-tubal junction during pre- and early postovulatory intervals. It was also higher (P < 0.05) than in the fluid from the ampullary-isthmic junction during pre- and early postovulatory intervals; however, sperm velocity in follicular fluid was not significantly different from that in the periovulatory fluid from the ampullary-isthmic junction. The mean lateral head displacement (ALHmean) of spermatozoa was significantly greater in follicular fluid (3.9 +/- 0.3 microns; n = 5) than in fluid from the ampulla during peri- and early postovulatory intervals and from the isthmic-utero-tubal junction during pre- and early postovulatory intervals, and was also higher (P < 0.05) than in fluid from the ampullary-isthmic junction during the preovulatory period, but was not different from the peri- and postovulatory ampullary-isthmic junction fluids. The proportion of spermatozoa exhibiting circular motion was significantly higher (P < 0.05) in the periovulatory fluid from the ampullary isthmic junction (24 +/- 3%) compared with fluids obtained during preovulatory and early postovulatory periods. Follicular fluid had no effect on the proportion of spermatozoa exhibiting circular motion. The average radius of sperm movement in circular trajectories was higher in follicular fluid than in the periovulatory fluids from the ampulla and ampullary-isthmic junction (P < 0.05). In hydrosalpingeal fluids collected 2-5 days after ovulation, the average radius of movement was greater in the ampulla fluid and ampullary-isthmic junction fluid than in fluid from the isthmic-utero-tubal junction (P < 0.05). These results demonstrate that follicular fluid and oviductal fluids have considerable influences on boar sperm motility. Furthermore, the immediate effect of periovulatory ampullary-isthmic junction fluid in increasing the percentage of spermatozoa swimming in circles (hyperactivated) is relevant, since it is at this time and within this region that fertilization occurs. PMID- 9227361 TI - Effects of mating dynamics and crowding on sex ratio variance in mice. AB - Mating units of six virgin females and one adult stud male were established to test for the effects of timing of mating and crowding of pregnant females on litter sex ratios in mice. Females either copulated during periods when no other female of the mating unit copulated simultaneously (single mating condition) or when more than one female copulated (multiple matings condition). Two crowding conditions were imposed on the animals: the females of 14 mating units were placed into individual cages after mating (isolated condition), while females of the other 13 mating units remained in the original group until shortly before littering (crowded condition). Sex ratio variance did not deviate from random expectation in litters arising from the multiple matings periods. However, in litters arising from single mating periods, extreme sex ratios were found significantly less frequently than expected by chance. Higher sex ratio variance in litters arising from multiple matings periods is attributed to the timing of mating being at higher variance under this condition, which is known to affect sex ratios in other rodents. Crowding significantly reduced sex ratio variance further. Reduced sex ratio variance under single mating and crowded conditions is speculated to follow from competition for resources between preimplantation embryos, which may be further increased by stressful effects of crowding. Loss of embryos after implantation appeared not to be responsible for the above effects. PMID- 9227363 TI - Impact of different patterns of feed intake during lactation in the primiparous sow on follicular development and oocyte maturation. AB - The potential contribution of nutritionally induced differences in follicular and oocyte maturity to embryo survival was addressed in pigs. When primiparous, lactating sows are fed to appetite from farrowing to day 21 of lactation and then with feed intake restricted to 50% from day 22 to 28 (restricted), embryo survival is 64% at day 28 of gestation, compared with 85% in sows fed to 50% from farrowing to day 21 and then fed to appetite from day 22 to 28 (refed). In the present study, 32 sows were equally assigned to these two treatments (restricted or refed) but they were slaughtered 38 h before the estimated time of oestrus. The largest 15 follicles per sow were aspirated and follicular fluid recovered for analysis in vitro. Although plasma oestradiol concentration before slaughter and follicular fluid oestradiol concentration at slaughter were not different (P > 0.05), refed sows had more (P < 0.02) large follicles than did restricted sows. Cumulus expansion scores in vitro were not different between treatments, although more (P < 0.03) oocytes from refed sows had matured to metaphase II than those from restricted sows. Similarly, although cumulus expansion of oocyte-cumulus complexes from prepubertal gilts oocytes incubated with follicular fluid obtained from restricted (n = 1227) or refed (n = 1147) sows was not different (P > 0.05), the rate of oocyte nuclear maturation was greater (P < 0.012) after incubation with follicular fluid from refed than with that from restricted sows. Differences in the maturation of the follicle and oocyte in the period before the LH surge may therefore contribute to the treatment effects on embryo survival. PMID- 9227362 TI - Enhancement of ovarian follicle development in heifers by treatment with recombinant bovine somatotrophin: a dose-response study. AB - Treatment with recombinant bovine somatotrophin (bST) can enhance the development of ovarian antral follicles in cattle. The underlying mechanism was examined further by performing a dose-response study to investigate the effects of bST on peripheral concentrations of somatotrophin, insulin-like growth factor I (IGF-I), insulin, FSH and LH, and ovarian follicle development. Twenty mature heifers were randomly divided into five groups and injected s.c. at 6 h intervals for 7 days with 25% of one of the following daily doses of bST: 0, 3.13, 6.25, 12.5 or 25.0 mg. Ovarian follicular dynamics were monitored by real-time ultrasonography. Blood samples were collected daily during the experiment, and every 15 min for 8 h on days 1 and 5 of bST treatment. Treatment with bST increased (P < 0.01) peripheral concentrations of somatotrophin in a dose-dependent manner. Serum concentrations of both IGF-I and insulin were significantly (P < 0.01) increased in all heifers given 12.5 or 25.0 mg bST per day. Peripheral concentrations of IGF-I and insulin in all animals in the group given 3.13 mg bST and two heifers in the group given 6.25 mg bST were not different from those in the control group, while concentrations in the other two heifers given 6.25 mg bST were significantly (P < 0.01) higher. The number of ovarian follicles < 5 mm in diameter was increased (P < 0.05) in response to bST, but only in heifers (n = 10) with significantly increased serum concentrations of IGF-I and insulin. There were no effects of treatment on peripheral concentrations of FSH, LH and progesterone, and on the numbers of follicles > 5 mm in diameter. In conclusion, this study has demonstrated in vivo that the effect of treatment with bST on ovarian follicle development appears to be mediated through an increase in circulating IGF-I or insulin concentrations, rather than via an alteration in the secretion of pituitary gonadotrophins or a direct effect of bST on ovarian follicles. PMID- 9227364 TI - Control of extracellular matrix remodelling within ovarian tissues: localization and regulation of gene expression of plasminogen activator inhibitor type-1 within the ovine corpus luteum. AB - Extensive extracellular matrix remodelling occurs within the lifespan of the corpus luteum, particularly during corpus luteum formation and regression. A major mechanism for the regulation of extracellular matrix remodelling is via local production of specific proteinase inhibitors, such as the serine proteinase inhibitor plasminogen activator inhibitor type-1 (PAI-1). The objective of the present study was to characterize the localization, ontogeny and regulation of PAI-1 expression within ovine corpora lutea. Urokinase binding activity was detected within medium conditioned by ovine luteal cells. Production of PAI-1 by ovine luteal cells was confirmed by immunoprecipitating it from labelled proteins in culture medium. mRNA encoding PAI-1 was present within developing (day 3), mature (day 10) and regressing (30 h after prostaglandin F2 alpha injection on day 10 after the onset of oestrus) corpora lutea as demonstrated by in situ hybridization. The ontogeny of PAI-1 mRNA expression was characterized within corpora lutea collected on days 3, 7, 10, 13 and 16 after the onset of oestrus (n = 4, 4, 4, 3 and 4, respectively). Expression of PAI-1 mRNA did not differ during the luteal phase (P = 0.06), although a trend for an increase in the amount of PAI-1 mRNA was observed on day 16. Expression of PAI-1 mRNA was also examined during luteal regression in corpora lutea collected 0, 6, 12, 24 and 36 h after injection of prostaglandin F2 alpha on day 10 after the onset of oestrus (n = 4 at each time). Relative PAI-1 mRNA concentrations changed significantly during luteolysis induced by prostaglandin F2 alpha (P = 0.0002). Administration of prostaglandin F2 alpha resulted in a transient sevenfold increase in PAI-1 mRNA 6 h after injection (P = 0.0001) but by 12 h the amounts had returned to values similar to those detected on day 10. We conclude that PAI-1 is a major secretory product of ovine luteal cells and that a transient increase in PAI-1 mRNA occurs during luteolysis induced by prostaglandin F2 alpha. PAI-1 probably plays a key local role in the control of extracellular proteolysis during the luteal phase. PMID- 9227365 TI - Ovulation induction and gamete transport in the female tract of the musk shrew, Suncus murinus. AB - The musk shrew Suncus murinus was studied with regard to induction of ovulation, the genesis and role of the vaginal copulation plug, and the behaviour of gametes and embryos within the Fallopian tube. Ovulation occurred about 15 h after ejaculation, which required a mean of 5.2 (range 2-10) intromittent thrusts. Since ovulation occurred also after five thrusts without ejaculation, and after ejaculation without plug formation or sperm deposition, the primary stimulus for this seemed to be the thrust of the penis, the glans of which was covered by a dense field of spines. Neither vasectomized nor prostatectomized males formed a plug at ejaculation, and in the latter case the mean number of spermatozoa reaching the isthmus of the Fallopian tube, the number at the ampullary fertilization site and the rate of fertilisation were lower than in females mated to normal males. Thus both the vesicular gland on the vas deferens and the prostate are essential for formation of the copulation plug, which appears to enhance sperm transport within the female tract. At ejaculation, < or = 10(6) spermatozoa were incarcerated by the plug in the anterior vagina for 6-7 h, by which time a maximal population of several hundred had become established in posterior crypts of the isthmus of the Fallopian tube as small groups of free languidly moving spermatozoa. It remains to be established whether oviductal crypts in this and other shrews have a storage function for spermatozoa or sequester spermatozoa and so regulate the number that reach the fertilization site. Very few spermatozoa reached the ampulla of Suncus. Generally, only one or two spermatozoa had reached the ampulla by 4-5 h, and often less than ten had done so by 5-13 h after ovulation. As a probable correlate, few eggs were penetrated during the first 5 h, with a frequent delay of 10-13 h before most eggs were fertilized. Thereafter, unfertilized eggs were transported through the oviduct at the same rate as developing embryos, which entered the uterus about 85 h after ovulation at the 32-cell stage. There were highly significant differences between the larger KAT/SK strains and smaller OK strain with regard to Fallopian tube length (mean 6.9 mm versus 9.7 mm), as well as the rates of hCG-induced ovulation (5.6 versus 3.25) and of unilateral ovulation (6% versus 50%). PMID- 9227366 TI - The unusual state of the cumulus oophorus and of sperm behaviour within it, in the musk shrew, Suncus murinus. AB - In the musk shrew, Suncus murinus, the behaviour of the cumulus-egg complex and its interaction with spermatozoa were unusual in several respects. The cumulus oophorus was ovulated about 15.5 h after mating or treatment with hCG as a hyaluronidase-insensitive matrix-free ball of cells which remained for relatively long periods of about 14 h around fertilized, and for about 24 h around unfertilized eggs. As a probable function of the small number of up to about 10 or 20 spermatozoa that generally reached the oviduct ampulla from isthmic crypts, there was often a delay of up to 10 h after ovulation before most eggs were penetrated. Soon after ovulation, however, the corona radiata retreated progressively from the zona pellucida, creating a closed perizonal space within the cumulus oophorus. Usually, most spermatozoa that did reach the ampulla were found within a cumulus and generally within that perizonal space. However, whereas the acrosome was intact among the few free ampullary spermatozoa, and in those adhering to the zona of cumulus-free eggs after delayed mating, all spermatozoa seen moving within the cumulus or adhering to the zona of unfertilized eggs had shed the giant acrosome. In accord with current observations in other shrews, the cumulus in Suncus may therefore function not only to sequester spermatozoa, but also as an essential mediator of fertilization probably by inducing the acrosome reaction. In the absence of the acrosomal carapace that expresses the zona receptors in most mammals, fertilizing Suncus spermatozoa could use an unusual array of barbs on the exposed perforatorium to attach to the zona pellucida. PMID- 9227367 TI - Migration of primordial germ cells to the developing gonadal ridges in the tammar wallaby Macropus eugenii. AB - Primordial germ cells (PGCs) of the tammar wallaby Macropus eugenii have a distinctive morphology and stain positively for alkaline phosphatase. PGCs are identifiable in embryos with 12 somites, on about day 17 of the 26.5 day gestation period, when they are located in all three germ layers of the developing embryo and in the endoderm of the bilaminar and vascular (trilaminar) yolk sac membranes. PGCs are positive for alkaline phosphatase (ALP) at least between days 17 and 22 of pregnancy. In whole mounts on day 17, three groups of cells positive for ALP occur: about 40 just caudal to the neural tube, and about 20 distributed on either side of the last three somites. By day 21, there are about 150 PGCs in the newly formed gonadal ridges and 275 in the mesenteries. On days 21-22, there are PGCs in the umbilical mesoderm, the dorsal mesentery and the coelomic angles between the dorsal mesentery and the mesonephroi. On day 22, most ALP-positive PGCs are located in the dorsal mesentery, where they occur in groups. They apparently do not migrate through the hindgut endoderm, but occasional PGCs are seen in sites such as the mesonephros, the adrenals, the blood vessels of the yolk sac and in the vicinity of the dorsal aorta and dorsal nerve cord. Between day 23 and day 25, 1 day before birth, most of the 3200-4000 PGCs complete their migration to the gonadal ridges. Although there are marked differences between embryogenesis of tammars and mice, development and the pattern of migration of PGCs in this marsupial mammal are similar to that of eutherian mammals. PMID- 9227368 TI - Melatonin receptors in red deer fetuses (Cervus elaphus). AB - Red deer (Cervus elaphus) exhibit highly seasonal rhythms in physiology and behaviour. The influence of photoperiod on the timing of these changes begins in utero where the fetus receives photoperiodic information via the diurnal pattern of maternal melatonin secretion. The potential sensitivity of deer fetuses to melatonin was ascertained by mapping specific receptors and characterizing them using 2-[125I]iodomelatonin and quantitative autoradiography in vitro. Specific binding occurred from day 31 of gestation onwards (term = 233 days) over the spinal nerves and respiratory system. At later stages of gestation binding occurred over the brain, particularly the brainstem, the pituitary gland, thyroid gland, gastrointestinal tract including the pancreas, metanephros, cochlea of the ear, spinal cord, and spinal and cranial nerves. Binding was abolished in the presence of 10(-7) mol melatonin l-1 and diminished in the presence of 10(-4) mol GTP gamma S l-1 (guanosine-5-O-(3-thiotriphosphate)), confirming that binding represented functional G-protein-coupled melatonin receptors. Characterization studies, carried out on fetal lung, revealed that binding was time-dependent, reaching equilibrium at about 3 h at room temperature (22 degrees C), and saturable with a dissociation constant (Kd) of 104 pmol l-1. This study demonstrates the presence of G-protein-coupled melatonin receptors over a wide range of tissues in red deer fetuses from early in gestation, indicating that in addition to its role in the communication of photoperiodic information to the fetus in this species, melatonin may be involved in fetal growth and development. PMID- 9227369 TI - Follicular growth pattern in cyclic rats from late pro-oestrus to early oestrus. AB - Adult cyclic rats were studied from 16:00 h on pro-oestrus to 07:00 h on oestrus to relate the cyclic hormonal changes to the proliferative activity and growth pattern of growing follicles. The proliferative activity was studied by 5 bromodeoxyuridine (BrdU) labelling and by the presence of mitoses. Small growing follicles (less than 275 microns in diameter) were divided into five classes: multilaminar classes a (Ma, up to 75 microns in diameter), b (Mb, 76-150 microns), c (Mc, 151-200 microns) and d (Md, 201-274 microns) and follicles measuring > or = 275 microns in diameter were considered as > or = class 1, following previous classifications. I.H concentrations were maximal at 18:30 h on pro-oestrus, and this was coincident with an increase in FSH, prolactin and progesterone concentrations, whereas oestradiol and testosterone concentrations were decreased. From 02:00 h on oestrus the concentrations of all hormones, except those of FSH, were decreased. The number of Ma, Mb and Mc follicles did not change during pro-oestrus-oestrus, whereas an increase in the number of follicles > or = class 1 was found at 07:00 h on oestrus. This appears to be a consequence of the increased proliferative activity of Md follicles, evidenced by the increase in the BrdU labelling and mitotic index of this follicle class, found from 02:00 to 07:00 h on oestrus, together with a decrease in the percentage of early atretic follicles > or = class 1 at 07:00 h on oestrus. This study provides an improved classification of small growing follicles into discrete classes and delineates a size class of follicles (Md follicles) that is responsive to the cyclic hormonal changes on early oestrus. PMID- 9227370 TI - Interstitial orchitis with impaired steroidogenesis and spermatogenesis in the testes of rabbits infected with an attenuated strain of myxoma virus. AB - The pathogenesis of infertility associated with acute viral orchitis was investigated in a rabbit model. Infection of postpubertal male European rabbits with an attenuated strain of myxoma virus caused systemic disease with viral replication to high titres in the testes. Infected rabbits developed an interstitial orchitis and epididymitis. This was associated with degeneration of the seminiferous epithelium, decreased serum testosterone concentrations and increased serum LH concentrations. Virus was localized within the interstitial cells. Clearance of infectious virus from the testis occurred between day 20 and day 30 after infection, although viral DNA could still be detected by polymerase chain reaction at 120 days. Viral clearance was associated with a return to normal serum testosterone and LH concentrations. Anti-sperm antibodies were present in the serum as early as 5 days after infection and declined during the recovery phase. Rabbits were infertile at 60 days but returned to normal fertility 60-90 days after infection. PMID- 9227371 TI - Regulation of mouse epididymal epithelium in vitro by androgens, temperature and fibroblasts. AB - The epididymal epithelium provides the microenvironment for sperm maturation. However, the molecular basis of epididymal function is still poorly understood because of the limitations of in vivo systems. For this reason, we have developed an in vitro culture system for mouse epididymal epithelial cells. Cells were purified by enzymatic digestion and centrifugation through a Percoll gradient, and plated on inserts coated with a replacement basement membrane. Cultured cells maintained ultrastructural and immunocytochemical features of epithelia, but did not retain the androgen responsiveness of epididymal cells (as judged by androgen receptor detection and secretion of specific markers) unless cocultured with fibroblasts. The androgen receptor was detected in the nuclei of epididymal epithelial cells only when grown with epididymal fibroblasts in the subjacent chamber. Moreover, specific epididymal secretory proteins were secreted only when epithelial cells were cultured in the presence of both androgens and fibroblasts at 32 degrees C. These results highlight the importance of cell-cell interaction, as well as temperature regulation in the physiology of the epididymis. They also establish the existence of two independent pathways in the differentiation of these cells. The first, leading to the expression of epithelial characteristics, is fibroblast-independent, whereas the second, conferring tissue-specific features, depends upon coculture with fibroblasts. PMID- 9227372 TI - Immunocytochemical and quantitative study of actin, desmin and vimentin in the peritubular cells of the testes from elderly men. AB - A quantitative immunohistochemical study using light and electron microscopy was carried out to evaluate the morphological and quantitative distribution of the peritubular cells that immunoreact with actin, vimentin and desmin, alone or in combinations, in normal adult testes and the changes in these cells in elderly men. Seminiferous tubules in ageing testes were classified in three groups according to the degree of lamina propria thickening due to tubular sclerosis: group I, < 8 microns; group II, 8.1-12 microns; and group III, > 12.1 microns. The number of peritubular cells per cross-sectioned tubule increased from group I to group III tubules. However, no significant differences between ageing men and controls were found in the total number of peritubular cells per testis. Most peritubular cells of control testes and of group I and group II tubules displayed immunoreactivity to actin. The peritubular cells in the outermost layers of group III tubules showed no or scanty reaction. The number of actin-immunostained cells per cross-sectioned tubule decreased (P < 0.05) with tubular sclerosis. The total number of these cells per testis was significantly lower (P < 0.05) in elderly men. A narrow band around the seminiferous epithelium immunostained for desmin in control testes and group I tubules. These cells also immunoreacted to actin and vimentin. In group II and, principally, in group III tubules, only isolated peritubular cells were immunostained for desmin. The number of desmin immunostained cells per cross-sectioned tubule decreased with tubular sclerosis and the total number per testis was also lower in elderly men. Vimentin immunostaining was observed in most peritubular cells in all tubule groups; these cells also immunoreacted to actin. Vimentin and desmin co-localized only in the inner peritubular cell layers. The number of vimentin-immunostained cells per cross-sectioned tubule increased with the degree of tubular sclerosis but the total number of these cells per testis did not differ significantly between control and ageing testes. PMID- 9227373 TI - Swearing an oath. PMID- 9227374 TI - Which doctor? The patient's view. PMID- 9227375 TI - Problem setting and problem solving: the role of evidence-based medicine. PMID- 9227377 TI - Ageing of bruising in children. PMID- 9227376 TI - Angiogenesis and the heart: therapeutic implications. PMID- 9227378 TI - Renovascular disease: the fifth frontier. PMID- 9227379 TI - Complications of 1303 central venous cannulations. AB - Central venous catheterization (CVC), now a common procedure, has several major complications. We assessed their incidence in a prospective study of 1303 cannulations done in the intensive care unit or operating theatre. Chest radiographs were obtained to verify proper catheter placement and to detect pneumothorax. Complications were arterial puncture in 68 (5.2%) patients, arrhythmias in 21 (1.6%), cardiopulmonary arrest in 1 (0.1%), and pneumothorax in 5 (0.5%). The tip of the CVC was incorrectly located in 149 (11.2%). The chest radiograph was a valuable method for detecting complications of central venous catheterization. PMID- 9227380 TI - Assault patients attending a Scottish accident and emergency department. AB - Over 2 months in 1995, 235 assault patients attended the accident and emergency department of the Royal Alexandra Hospital, Paisley (2.4% of total new attendances). 80% were male and their mean age was 28 years (range 6-64); men were the assailants in over 90% of attacks. Alcohol had been consumed by 69% of the victims and 9% admitted to taking illicit drugs. The commonest place of assault was the street (44%) but women were more likely to be assaulted in their homes. Penetrating weapons were used in 23% of assaults. 60% of all injuries were to the head and neck. 27% of the victims were admitted to hospital. Paisley has an assault rate similar to that of other UK centres but the use of penetrating weapons is much higher than elsewhere. PMID- 9227381 TI - Psychiatric dilemmas--surgery and the Mental Health Act (1983). AB - Clinicians commonly face an ethical conundrum when the patient declines a treatment they believe to be necessary. The decision, whether to give or to withhold treatment, may also have legal repercussions. Two illustrative cases are presented. Although the Mental Health Act (1983) is concerned with treatment of mental disorder, treatments for physical disorders may be given in certain circumstances if the patient is deemed to lack the competence to understand the issues involved in the decision, and treatment is deemed to be in their best interests. PMID- 9227382 TI - Development of an essential drugs list for Bosnia and Herzegovina. AB - Part of the impact of the war in ex-Yugoslavia and especially Bosnia and Herzegovina was to limit the supply of therapeutic drugs they had used before the war. The difficulties encountered made the health care system temporarily dependent on humanitarian assistance agencies which applied the concept of essential drugs; and, after initial difficulties, national health staff adapted to the need to prescribe from a very limited range of drugs. Meanwhile, national drug policy and procurement and prescribing practices were reviewed by working groups and a national List of Essential Drugs was drawn up by national experts with international support. This list has now been passed into legislation. PMID- 9227383 TI - Primary hydatid cyst mistaken for carcinoma of the pancreas. PMID- 9227384 TI - Head injuries in infants caused by falls from surfaces while restrained in car seats. PMID- 9227385 TI - Chronic parathyroiditis associated with primary hyperplastic hyperparathyroidism. PMID- 9227386 TI - Stromal cell tumour of rectum treated by transanal endoscopic microsurgery. PMID- 9227387 TI - Small-cell carcinoma of the prostate. PMID- 9227388 TI - Disease and dictatorship: the case of Hitler's Reich. PMID- 9227389 TI - On mentoring. PMID- 9227390 TI - Hughlings Jackson comes home. PMID- 9227391 TI - Hazards of growing up. PMID- 9227392 TI - Paradigm shifts in the 16th and 17th centuries. PMID- 9227393 TI - Complementary medicine in the medical curriculum. PMID- 9227394 TI - Complementary medicine in the medical curriculum. PMID- 9227395 TI - Complementary medicine in the medical curriculum. PMID- 9227397 TI - Evolutionary psychiatry. PMID- 9227396 TI - Intraluminal stenting in the management of adhesional intestinal obstruction. PMID- 9227398 TI - Iliopectineal bursitis. PMID- 9227399 TI - Helicobacter pylori and cancer. PMID- 9227400 TI - Hypertension management in general practice. PMID- 9227402 TI - T-type and L-type Ca2+ currents in canine bronchial smooth muscle: characterization and physiological roles. AB - We examined the voltage-dependent Ca2+ currents in freshly dissociated smooth muscle cells obtained from canine bronchi (3rd to 5th order). When cells were depolarized from -40 mV, we observed an inward current that 1) exhibited threshold and peak activation at approximately -35 mV and +10 mV, respectively; 2) inactivated slowly with half-inactivation at -20 mV; 3) deactivated rapidly (time constant < 1 ms) upon repolarization; and 4) was abolished by nifedipine and suppressed by cholinergic agonist. These characteristics are typical of L type Ca2+ current. During depolarization from -70 or -80 mV, however, many cells exhibited a second inward current superimposed on the L-type Ca2+ current. Activation of this other current was first noted at -60 mV, was maximal at approximately -20 mV, and was very rapid (reaching a peak within 10 ms). Inactivation of the other current was also rapid (time constant approximately 3 ms) and half-maximal at approximately -70 mV. There was a persistent "window" current over the physiologically relevant range of potentials (i.e., -60 to -30 mV). This current was also sensitive to nifedipine (although less so than the L type current) and to Ni2+, but not to cholinergic agonist. Finally, the tail currents evoked upon repolarization to the holding potential decayed approximately 10 times more slowly than did L-type tail currents. These characteristics are all typical of T-type Ca2+ current. We conclude that there is a prominent T-type Ca2+ current in canine bronchial smooth muscle; this current may play a central role in excitation-contraction coupling, in refilling of the internal Ca2+ pool, and in electrical slow waves. Because airflow resistance is determined primarily by the smaller airways and not the trachea, these findings may have important implications with respect to airway physiology and the mechanisms underlying airway hyperreactivity and asthma. PMID- 9227403 TI - Intracellular electrolytes regulate the volume set point of the organic osmolyte/anion channel VSOAC. AB - Regulation of the volume sensitivity of the swelling-activated organic osmolyte/anion channel VSOAC by intracellular electrolytes was examined in intact and patch-clamped C6 glioma cells. In intact cells, VSOAC activation was monitored by [3H]taurine efflux measurements, and intracellular electrolyte concentrations were manipulated by acclimation to hypertonic medium for varying periods of time. Hypertonic shrinkage was followed by a rapid and complete regulatory volume increase mediated by electrolyte accumulation that elevated intracellular Na+, K+, and Cl- concentrations. During prolonged (4-48 h) exposure to hypertonicity, electrolyte concentrations decreased gradually as cells accumulated the organic osmolyte myo-inositol. VSOAC activation induced by cell swelling of 35-40% increased as a function of the time of exposure to hypertonicity and was inversely correlated with measured intracellular Na+, K+ and Cl- levels. In patch-clamped cells, swelling-induced Cl- current activation was unaffected by acclimation conditions but was inhibited by increasing the concentration of electrolytes in the patch pipette solution. Quantification of the relationship between VSOAC activation and cell swelling demonstrated that increases in intracellular electrolyte levels increase VSOAC volume set point. Regulation of VSOAC volume sensitivity by electrolytes allows cells to selectively utilize electrolytes or a combination of electrolytes and organic osmolytes for regulatory volume decrease (RVD). Control over the type of solute used for volume regulation is advantageous, allowing cells to control intracellular ionic composition and prevent increases in cytoplasmic ionic strength during RVD. PMID- 9227405 TI - Delivery of newly synthesized Na(+)-K(+)-ATPase to the plasma membrane of A6 epithelia. AB - Na(+)-K(+)-ATPase is localized to the basolateral cell surface of most epithelial cells. Conflicting results regarding the intracellular trafficking of Na(+)-K(+) ATPase in Madin-Darby canine kidney cells have been reported, with delivery to both apical and basolateral membranes or exclusively to the basolateral cell surface. We examined the delivery and steady-state distribution of Na(+)-K(+) ATPase in the amphibian epithelial cell line A6 using an antibody raised against Na(+)-K(+)-ATPase alpha-subunit and sulfo-N-hydroxysuccinimidobiotin to tag cell surface proteins. The steady-state distribution of the Na(+)-K(+)-ATPase was basolateral, as confirmed by immunocytochemistry. Delivery of newly synthesized Na(+)-K(+)-ATPase to the cell surface was examined using [35S]methionine and [35S]cysteine in a pulse-chase protocol. After a 20-min pulse, the alpha-subunit and core glycosylated beta-subunit were present at both apical and basolateral cell surfaces. The alpha-subunit and core glycosylated beta-subunit delivered to the apical cell surface were degraded within 2 h. Mature alpha/beta-heterodimer was found almost exclusively at the basolateral surface after a 1- to 24-h chase. These data suggest that immature Na(+)-K(+)-ATPase alpha-subunit and core glycosylated beta-subunits are not retained in the endoplasmic reticulum of A6 cells and apparently lack sorting signals. Mature Na(+)-K(+)-ATPase is targeted to the basolateral surface, suggesting that basolateral targeting of the protein is conformation dependent. PMID- 9227404 TI - Ca2+ flux in human placental trophoblasts. AB - Intracellular Ca2+ is an important second messenger. In the placenta, regulation of intracellular Ca2+ concentration ([Ca2+]i) by extracellular factors has received relatively little attention. Cultured human placental trophoblasts were treated with a series of potential Ca(2+)-mobilizing ligands. After 3 days in culture, there was an increase in [Ca2+]i in response to angiotensin, endothelin, transforming growth factor-alpha, and ATP in approximately 8, 54, 17, and 100% of the cells, respectively. The response to ATP was dose dependent. At low ATP concentrations (1-10 microM), the response to repeat ATP application remained unchanged, whereas at 100 microM, response to repeat stimulation resulted in lower peak value. The order of potency for the ATP derivatives was ATP = UTP > benzoylbenzoic-ATP > ATP gamma S > ADP beta S > ADP > alpha,beta-MeATP > AMP. This suggests action via the P2u purinergic receptor. Removal of extracellular Ca2+ decreased the ATP-induced Ca2+ response by 45%; this indicates that a substantial portion of the increase in [Ca2+]i was due to influx from extracellular space. Finally, ATP rapidly induced inositol 1,4,5-trisphosphate formation in cultured trophoblasts. Therefore, ATP-induced changes in Ca2+ flux may be due in part to activation of phosphoinositide-specific phospholipase C. In summary, ATP is a potent calciotropic ligand in human placental trophoblasts, acting through the P2u receptor. The effect of ATP on [Ca2+]i may prove to be involved in the modulation of various trophoblast functions, including hormone secretion and active transport of nutrients. PMID- 9227406 TI - Regulation of inducible and neuronal nitric oxide synthase gene expression by interferon-gamma and VIP. AB - The studies examined the regulation of inducible and neuronal nitric oxide synthases (iNOS and nNOS, respectively) in the rat brain, stomach, rectum, and spleen. Relative expression of iNOS and nNOS mRNAs was quantified by the sensitive method of polymerase amplification reactions. The NOS proteins were determined by Western blot, using specific antibodies. Highest levels of nNOS and iNOS mRNAs were expressed in the rat brain and spleen, respectively. Furthermore, both nNOS and iNOS were expressed in the stomach and rectum. Interestingly, treatment of tissues with lipopolysaccharides or cytokine interferon-gamma (IFN gamma) induced the expression of iNOS and decreased that of nNOS, representing a shift from one isoform to the other. When the tissues were treated with IFN-gamma followed by vasoactive intestinal polypeptide (VIP), the induction of iNOS was reduced by VIP. The changes in iNOS and nNOS expression at the transcriptional level corresponded to those at the translational level. The data suggest a regulatory role of IFN-gamma and VIP in the expression iNOS and nNOS and a counterregulation of iNOS and nNOS in rat tissues. PMID- 9227407 TI - Volume regulation in NIH/3T3 cells not expressing P-glycoprotein. I. Regulatory volume decrease. AB - Exposure of NIH/3T3 fibroblasts not expressing P-glycoprotein to 50, 30, 20, and 10% hyposmotic solutions led to cell volume increases of 70, 32, 21, and 12%, respectively. After swelling, NIH/3T3 cells exhibited regulatory volume decrease (RVD), attaining complete volume recovery after 30 min except in 50% hyposmotic solution, in which volume recovery was 76%. RVD was accelerated by gramicidin and inhibited by the Cl channel blockers 5-nitro-2-(3-phenylpropylamino)-benzoic acid, 1,9-dideoxyforskolin, dipyridamole, and niflumic acid and by the K channel, blocker quinidine. RVD was reduced 15% by removal of extracellular Ca. The pathway opened by hypotonicity was highly permeable to K and Rb and only partly permeable to other cations. Most anions were able to permeate, with a permeability ranking of nitrate > benzoate = iodide > thiocyanate > chloride > > gluconate. The pathway was permeable to neutral amino acids, with a permeability ranking of glycine > alanine > glutamate > taurine > gamma-aminobutyric acid > glutamine. The pathway was not permeable to basic amino acids. These results show that, despite the absence of P-glycoprotein, NIH/3T3 cells exhibit RVD with properties similar to those expressed in most cell types. PMID- 9227409 TI - Heterologous desensitization of smooth muscle to K+ depolarization: retarded stimulus-[Ca2+]i coupling. AB - To understand the phenomenon of postreceptor heterologous desensitization, we exposed porcine carotid media to 40 mM KCl physiological saline solution both before and after intervening treatment with histamine. Increasing histamine concentration or duration of exposure or decreasing the interval between histamine exposure and KCl progressively slowed the contractile responses to K+ depolarization. A delay in initiation and a slower rate of rise of KCl-induced stress in histamine-pretreated muscle were preceded by a slower rate of rise of aequorin-estimated myoplasmic Ca2+ concentration ([Ca2+]i), myosin regulatory light chain (MRLC) phosphorylation, and tissue stiffness, with no detectable change in the Ca2+ sensitivity of MRLC phosphorylation. This heterologous desensitization was not a diminished steady-state force but instead a profound slowing of contraction rates. This slowing was a manifestation of retardation of the rate at which [Ca2+]i rises to the level appropriate for the stimulus. The lack of rapid initial [Ca2+]i and cross-bridge phosphorylation transients as a consequence of histamine pretreatment resulted in very slow cross-bridge cycling rates and rates of force development (latch). PMID- 9227408 TI - Volume regulation in NIH/3T3 cells not expressing P-glycoprotein. II. Chloride and amino acid fluxes. AB - The osmolyte function of amino acids and Cl in native NIH/3T3 cells not expressing the P-glycoprotein was examined by investigating the free amino acid concentration and the swelling-activated efflux of [3H]taurine, as representative of amino acids, and of 125I, as a tracer for Cl. Taurine and 125I efflux was activated by 20 and 30% hyposmotic solutions. At 50% hyposmotic solutions, the osmolyte pool was essentially depleted. The Cl channel blockers 5-nitro-2-(3 phenylpropyl-amino)benzoic acid, 1,9-dideoxyforskolin, dipyridamole, and niflumic acid inhibited the release of the two osmolytes by 80-95%. 4,4' Diisothiocyanostilbene-2,2'-disulfonic acid (400 microM) decreased the efflux of taurine 80% without affecting that of 125I. Linolenic and arachidonic acids (5-20 microM) showed a concentration-dependent inhibitory effect on taurine and 125I fluxes. Omission of Ca decreased osmolyte fluxes by 16%. Verapamil inhibited the osmolyte release only at 500 microM. Nimodipine at 25 and 50 microM decreased the release of [3H]taurine and 125I by approximately 60 and 80%, respectively, but this effect was independent of the presence of extracellular Ca. These results indicate that amino acids and Cl function as osmolytes during regulatory volume decrease in native NIH/ 3T3 cells. PMID- 9227410 TI - Eicosanoid production from porcine neutrophils and platelets: differential production with various agonists. AB - Polymorphonuclear neutrophils (PMN) and platelets interact to produce both inflammatory and anti-inflammatory lipid mediators during human disease. Because swine models of human disease are used, it is important to understand the mechanisms involved in the formation of lipid mediators from porcine PMN-platelet interactions. In the present study, we investigated the mechanism of thromboxane (Tx) A2 and lipoxin A4 (LXA4) formation from porcine PMN and platelets, respectively. PMN (10(7)/ml) and platelet (30 x 10(7)/ml) suspensions stimulated with porcine C5a (pC5a), but not recombinant human C5a (rhC5a), significantly enhanced TxB2 formation. After cytochalasin B treatment, pC5a or rhC5a significantly and equally enhanced TxB2 formation from PMN-platelet suspensions. A-23187-induced TxB2 formation from platelets was not significantly augmented by the presence of PMN in these suspensions. A-23187 induced significant LXA4 production from porcine PMN that was not augmented by addition of platelets. Flow cytometric analysis of PMN-platelet suspensions revealed activated platelets adherent to PMN following pC5a stimulation. CV-6209, a platelet-activating factor (PAF) receptor antagonist, dose dependently prevented pC5a-induced platelet adherence to PMN and TxB2 formation. These data demonstrate that 1) porcine PMN alone can biosynthesize LXA4 without the assistance of platelets, which is in sharp contrast to human PMN-platelet interactions, and 2) in the absence of cytochalasin B, pC5a stimulates PAF biosynthesis from porcine PMN, resulting in TxB2 formation from platelets. PMID- 9227411 TI - IL-6 secreted by astroglial cells regulates Na-K-Cl cotransport in brain microvessel endothelial cells. AB - We showed previously that cerebral microvessel endothelial cells (CMEC) exhibit a prominent Na-K-Cl cotransporter that functions to regulate intracellular volume and may also mediate vectorial ion transport across the blood-brain barrier (BBB). Astrocytes and their conditioned media induce BBB properties of the endothelium. Our previous studies demonstrated that exposure of CMEC to astroglial cells markedly increases cotransport activity, upregulates cotransporter protein expression, and also increases cotransporter protein phosphorylation. In the present study, we evaluated the possibility that astroglial effects on the Na-K-Cl cotransporter are mediated by astroglial cell secreted interleukin-6 (IL-6). Cotransporter activity was assessed as bumetanide sensitive K influx, and protein expression was evaluated by Western blot analysis. Exposing CMEC to IL-6 for 48 h caused a dose-dependent stimulation of Na-K-Cl cotransport activity. Both C6 glial cell-conditioned medium (C6CM) induced and IL-6-induced increases in cotransport activity were neutralized by anti-IL-6 antibodies. A 48-h exposure of cells to IL-6 or C6CM also resulted in increased cotransporter protein expression. Furthermore, using an enzyme-linked immunosorbent assay for IL-6, we found significant amounts of IL-6 in C6CM. These data suggest that astroglia-secreted IL-6 may mediate the observed astroglial cell effects on CMEC Na-K-Cl cotransport and further support the hypothesis that astrocytes participate in maintenance of cerebral ionic homeostasis by regulating Na-K-Cl cotransporter function at the BBB. PMID- 9227412 TI - EGF and TGF-beta regulate neutral endopeptidase expression in renal vascular smooth muscle cells. AB - We recently reported that neutral endopeptidase (NEP) expression on renal vascular smooth muscle cells (VSMC) was downregulated in the presence of serum. Here we examine the role of epidermal growth factor (EGF) and transforming growth factor-beta 1 (TGF-beta) in this downregulation and the consequences of the changes in NEP activity on their mitogenic effects. EGF inhibited NEP activity, whereas TGF-beta was stimulatory. Expression of the enzyme was studied by measuring the binding of [125I]RB-104, a specific NEP inhibitor, and the fluorescence intensity of NEP-labeled cells. Both parameters were decreased by EGF and were increased by TGF-beta. NEP mRNA expression in EGF-treated cells was reduced after 48 h. In contrast, it was increased in TGF-beta-treated cells. Interestingly, NEP inhibition influenced the mitogenic effect of EGF. Indeed, thiorphan, an NEP inhibitor, and an anti-NEP antibody decreased EGF-dependent [3H]thymidine incorporation and cell proliferation by approximately 50%. TGF-beta had no effect on VSMC growth. These results indicate that EGF but not TGF-beta participates in the downregulatory potency of serum on NEP expression in VSMC. They also demonstrate that the full effect of EGF on VSMC proliferation depends on intact NEP activity. PMID- 9227413 TI - Regulation of M-like K+ current, IKx, by Ca(2+)-dependent phosphorylation in rod photoreceptors. AB - An M-like K+ current (IKx) helps set the rod photoreceptor resting potential and accelerates the response to dim light. Recorded with ruptured-patch whole cell techniques, the amplitude of IKx diminished, and activation occurred at increasingly negative potentials as a function of time. In contrast, IKx was stable during nystatin perforated-patch recording. Stability during ruptured patch recording could be induced by raising the intracellular Ca2+ concentration ([Ca2+]i) or by including caffeine or D-myo-inositol 1,4,5-trisphosphate in the pipette. This Ca(2+)-induced stability of IKx was blocked by inhibitors of Ca2+/calmodulin-dependent protein kinases, such as W-7, KN-62, chelerythrine, or H-7. Okadaic acid, an inhibitor of protein phosphatases, maintained IKx stability even at low [Ca2+]i. The requirement for phosphorylation was demonstrated by depleting MgATP or by providing 5'-adenylylimidophosphate, a nonhydrolyzable analog of ATP, either of which blocked the Ca(2+)-induced stability of IKx. These observations show that phosphorylation regulates IKx and that a stimulus controlling this action is [Ca2+]i. Should [Ca2+]i change during light adaptation, changes in IKx might alter the resting potential and temporal response properties of rod photoreceptors. PMID- 9227414 TI - Cell volume regulation by the mouse zygote: mechanism of recovery from a volume increase. AB - Mouse zygotes regulate their volumes after cell swelling. This regulatory volume decrease (RVD) is rapid and complete. RVD in zygotes was inhibited by K+ or Cl- channel blockers, indicating the participation of such channels in volume recovery. The channels are separate entities, as indicated by the ability of the cation ionophore gramicidin to restore RVD when K+ channels are blocked but not when Cl- channels are blocked. Intracellular Ca2+ concentration increased with cell swelling. Nevertheless, RVD occurred normally in zygotes loaded with the Ca2+ chelator, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid, which prevented Ca2+ from increasing above its normal resting concentration. Thus an increase in intracellular Ca2+ is not necessary for zygote RVD; consistent with this, inhibitors of Ca(2+)-activated K+ channels had little or no effect on RVD. RVD in zygotes was also completely inhibited by millimolar amounts of extracellular ATP. ATP has been shown to inhibit current passed by the volume sensitive organic osmolyte-Cl- channel in other cells, and thus zygotes may have such a channel participating in RVD. PMID- 9227415 TI - Role of purinergic receptors in chloride secretion in Caco-2 cells. AB - Purinergic receptors play an important role in regulating Cl- secretion in epithelial cells. To explore further the role of these receptors in the intestine, we utilized the human intestinal epithelial cell line, Caco-2, grown on permeable membrane supports and assayed for Cl- secretion by measuring the short-circuit current (Isc). Stimulation of Isc by extracellular nucleotides could be detected by day 4 and increased by day 10 postseeding. The magnitude of stimulation of Isc at 10 microM in cells at day 10 was UTP > ATP > UDP > > 2 methylthioadenosine 5'-triphosphate (2-MeS-ATP) = ADP on the apical side and UTP = 2-MeS-ATP = ATP > ADP > > UDP on the basolateral side. Cross-desensitization studies suggested that two different receptors are expressed in the apical membrane, a P2U purinoceptor and a uridine nucleotide receptor. Two different receptors are also expressed in the basolateral membrane, a P2U receptor and another that reacts with both 2-MeS-ATP and ADP. This latter receptor has an unusual pharmacological profile, with a reactivity for 2-MeS-ATP > ADP but not for ATP. Responses to purinergic receptor agonists were inhibited by pretreatment with 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid-acetoxymethyl ester, thapsigargin, or quinine. Thus we suggest that an increase in intracellular Ca2+ and subsequent opening of Ca(2+)-activated K+ channel play a role in increasing driving force for Cl- to exit across the apical membrane. The role of the cystic fibrosis transmembrane conductance regulator as a Cl- exit pathway on the apical membrane was also established. PMID- 9227417 TI - Mismatch between myosin heavy chain mRNA and protein distribution in human skeletal muscle fibers. AB - The distribution of myosin heavy chain (MHC) isoforms was analyzed at the protein and mRNA levels in human skeletal muscle biopsies from young normal adult subjects. Using ATPase histochemical reactions, antibodies to fast- and slow-type MHCs, and in situ hybridization with probes specific for MHC-beta/slow, MHC-2A, and MHC-2X, we confirmed our previous results showing that most fibers contain either a single mRNA and isoprotein or a mixed 1/2A or 2A/2X phenotype with coexistence of two mRNAs and isoproteins. However, we also found a minor proportion of fibers showing a mismatch in the relative proportion of mRNA and protein, e.g., fibers containing MHC-2A mRNA but not the corresponding protein or fibers containing MHC-2A protein but not the corresponding transcript. These fibers were more frequent in biopsies obtained after a training or detraining period than before the training period. We propose that these fibers represent transitional fibers and that the relative content of each mRNA and isoprotein gives a clue as to the direction of change in MHC gene expression. PMID- 9227416 TI - Immunolocalization of the ubiquitin-protein ligase Nedd4 in tissues expressing the epithelial Na+ channel (ENaC). AB - The epithelial Na+ channel (ENaC) was previously shown to be expressed in several Na(+)- and fluid-absorbing epithelia, particularly those of the kidney, colon, and lung. We have recently identified the ubiquitin-protein ligase Nedd4 as an interacting protein with ENaC and demonstrated that Nedd4 binds by its WW domains to the proline-rich PY motifs of ENaC. These PY motifs were recently shown to be deleted/mutated in patients afflicted with Liddle's syndrome, a hereditary form of systemic renal hypertension. Such mutations cause elevated channel activity by an increase in channel number/stability at the plasma membrane and by increased channel opening. We then proposed that Nedd4, by regulating channel stability/ degradation, may be a suppressor of ENaC. To test whether Nedd4 is localized to those tissues/regions that express ENaC, we performed immunocytochemical analysis of rat Nedd4 (rNedd4) distribution in rat kidney, colon, and lung tissues. Our results show that, in the kidney, rNedd4 is primarily localized to the cortical collecting tubules and outer and inner medullary collecting ducts. These tubular segments were previously shown to express ENaC. The epithelium lining medullary calyxes was also intensely stained, and microvillar borders of proximal convoluted tubules expressed variable amounts of rNedd4. In the lung, rNedd4 was mainly expressed in the epithelia lining the airways, in the submucosal glands and ducts, and in the distal respiratory epithelium. These sites resemble the pattern of ENaC expression. In contrast, in the distal colon, rNedd4 was strongly expressed in the epithelia lining the crypts but not in the ENaC-expressing surface epithelium. Low-salt diet (to elevate serum aldosterone levels) had no effect on rNedd4 distribution in the kidney or colon. Thus Nedd4 is coexpressed and likely colocalizes with ENaC in specific regions within the kidney and lung but not in the colon. We speculate this difference in colocalization may reflect differences in the regulation of channel stability in those tissues. PMID- 9227418 TI - Isovolumetric regulation in a distal nephron cell line (A6). AB - The effects of gradually reducing the osmolality of the basolateral solution (pi b) were examined in a renal epithelial cell line (A6). pi b was linearly decreased with time from 260 to 140 mosmol/ kgH2O. Cell volume did not change when pi b was gradually decreased at dilution rates (D(r)) of 1-1.5 mosmol. kgH2O 1. min-1. Increasing D(r) to 3 or 6 mosmol. kgH2O-1. min-1 abolished this isovolumetric regulation (IVR). Replacing Cl- by NO3- or SCN- inhibited IVR markedly, whereas Br- substituted perfectly for Cl-. On the other hand, with all these anions, the regulatory volume decrease (RVD) was completely developed. Ba2+ (30 mM) markedly slowed down RVD but improved IVR. The discrepancies between RVD and IVR suggest that different mechanisms are used to control cell volume during gradual and shockwise hyposmotic perturbations. During gradual and shockwise reductions of pi b, cellular K+ content was reduced to the same extent; 86Rb efflux was only partially inhibited by Ba2+. The amount of intracellular K+ depletion could account for 70% of the cationic osmolyte loss, which suggests that K+ is the major cation excreted during both types of perturbations. PMID- 9227420 TI - Actions of sulfhydryl reagents on single ryanodine receptor Ca(2+)-release channels from sheep myocardium. AB - Effects of the reactive disulfides, 2,2'- and 4,4'-dithiodipyridine, on single cardiac ryanodine receptor (RyR) ion channels incorporated into lipid bilayers are reported. RyRs are activated within minutes of addition of the reactive disulfides (10(-7) to 10(-3) M) with an irreversible loss of channel activity after the activation at concentrations > or = 10(-4) M. This activation, followed by loss of activity, is seen over a wide range of cytoplasmic (cis) Ca2+ concentration between 10(-9) and 2 x 10(-2) M and occurs more rapidly with higher reactive disulfide concentrations or when RyRs are initially active at 10(-5) or 10(-3) M Ca2+. The reactive disulfides increase the channel open probability by introducing long components into the open time distributions, increasing the mean channel open time by up to 50-fold. Closed time distributions are not altered by the sulfhydryl reagents. The effects of the reactive disulfides are prevented by the reducing agents dithiothreitol and glutathione (1-10 mM). The results suggest that cysteine residues on the RyR complex can regulate the ion channel gating mechanisms. PMID- 9227419 TI - Whole cell Cl- conductances in mouse choroid plexus epithelial cells do not require CFTR expression. AB - Whole cell patch-clamp studies were performed with tissue isolated from the cystic fibrosis (CF) transgenic Cftrm1cam mouse, to determine whether anion currents in choroid plexus epithelial cells require the expression of cystic fibrosis transmembrane conductance regulator (CFTR). Inclusion of 0.25 mM adenosine 3',5'-cyclic monophosphate (cAMP) and 375 nM protein kinase A (PKA) in the pipette solution caused a significant activation of a Cl(-)-selective, inward rectifying conductance in cells from wild-type and CF mice. The small, outward currents observed in wild-type and CF animals, however, were not activated by cAMP-PKA. There were no significant differences in the size of currents between wild-type, heterozygote, and CF cells in the presence or absence of cAMP-PKA. A second whole cell conductance was activated when cells from wild-type mice were swollen. These volume-activated currents were Cl- selective and exhibited outward rectification. They were Ca2+ independent and ATP dependent and blocked by 4,4' diisothiocyanostilbene-2,2'-disulfonic acid and 5-nitro-2-(3 phenylpropylamino)benzoic acid. The volume-activated channels were also activated in CF mutant cells, and there was no significant difference in the size of the volume-activated currents between wild-type, heterozygote, and CF cells. It is concluded that CFTR neither contributes to the whole cell conductance nor regulates the other anion conductances in choroid plexus epithelial cells. PMID- 9227421 TI - Calcium transients and calcium homeostasis in adult mouse fast-twitch skeletal muscle fibers in culture. AB - Skeletal muscle fibers enzymatically dissociated from adult mouse flexor digitorum brevis muscles were maintained in culture for up to 8 days. After various times in culture, fibers were loaded with fura 2, and Ca2+ transients for trains of 1, 5, and 10 action potentials (100 Hz) triggered by external electrical stimulation were calculated from fluorescence ratio records corrected for noninstantaneous reaction of fura 2 with Ca2+. The decay rate constants of Ca2+ transients decreased with increasing stimulation duration, indicating a slowing of the Ca(2+)-removal properties with increased stimulation duration. After 6 days in culture, Ca2+ decay rate constants decreased dramatically for all stimulation durations and the differences in decay rate constants among 1, 5, and 10 pulses became smaller. Intracellular parvalbumin content measured by single fiber immunofluorescence decreased with time in culture in parallel with the decrease in the decay rate constant of Ca2+ transients. Our results suggest that there is a correlation between parvalbumin content and the decay rate constant of the Ca2+ transient. PMID- 9227422 TI - Protein prenylation is required for aldosterone-stimulated Na+ transport. AB - Aldosterone stimulation of transcellular Na+ flux in polarized epithelial cells is dependent on at least one transmethylation reaction, but the substrate of this signaling step is unknown. Because it is clear that the majority of cellular protein methylation occurs in conjunction with protein prenylation, we examined the importance of prenylation to aldosterone-stimulated Na+ transport in the A6 cell line. Lovastatin, an inhibitor of the first committed step of the mevalonate pathway, inhibits the natriferic effect of aldosterone but does not inhibit insulin-stimulated Na+ flux. The addition of a farnesyl group does not appear to be involved in aldosterone's action. Neither alpha-hydroxyfarne-sylphosphonic acid, an inhibitor of farnesyl:protein transferase, nor N-acetyl-S-farnesyl-L cysteine, an inhibitor of farnesylated protein methylation, inhibits the hormone induced increase in Na+ transport. In contrast, N-acetyl-S-geranyl-geranyl-L cysteine, an inhibitor of geranylgeranyl protein methylation, completely abolishes the aldosterone-induced increase in Na+ flux with no effect on insulin mediated Na+ transport or cellular protein content. These data indicate that methylation of a geranylgeranylated protein is involved in aldosterone's natriferic action. PMID- 9227423 TI - Blocking ATP-sensitive K+ channel during metabolic inhibition impairs muscle contractility. AB - The objectives of this study were to determine the metabolic conditions in which ATP-sensitive K+ channels (K+(ATP) channels) contribute to a decrease in force. Sartorius muscles of the frog Rana pipiens were subjected to a 60-min metabolic inhibition by exposing them to cyanide (2 mM) and iodoacetate (1 mM). Muscles were exposed to glibenclamide (100 microM) to block K+ATP channels either 60 min before or 8 or 18 min into metabolic inhibition. Resting potentials, action potentials, and membrane conductance were measured using intracellular microelectrodes. Tetanic and resting tension were measured with a force transducer. ATP, ADP, and phosphocreatine (PCr) were measured by high-pressure liquid chromatography. Glibenclamide completely blocked the shortening of action potential but only partially blocked the increase in membrane conductance. When glibenclamide was added 60 min before metabolic inhibition, the decrease in tetanic force was faster than in control muscle (no glibenclamide). This faster decrease in tetanic force was associated with significant membrane depolarizations, greater increases in resting tension, greater depletions of ATP and PCr contents, and greater increases in ADP content. Addition of glibenclamide 8 min into metabolic inhibition caused an increase in tetanic force followed by a faster decrease compared with control. Addition of glibenclamide 18 min into metabolic inhibition had no effect on the tetanic force compared with control muscles. The data indicate that K+ATP channels 1) were activated during metabolic inhibition and 2) contributed to the decrease in tetanic force but also 3) had a myoprotective effect protecting skeletal muscle against muscle function impairment. PMID- 9227424 TI - Regulation of vascular smooth muscle cell proliferation by plasma membrane Ca(2+) ATPase. AB - We have previously shown that reductions in c-Myb-dependent transcription inhibit cell cycle progression and decrease intracellular Ca2+ concentrations in vascular smooth muscle cells (VSMC). We now report that these effects are largely mediated by a 4- to 10-fold increased rate of La(3+)-sensitive 45Ca extrusion, which is associated with 2- to 4-fold increased levels of plasma membrane Ca(2+)-ATPase 1 (PMCA1) mRNA and protein. PMCA4 mRNA, present at much lower concentrations, undergoes similar changes during suppression of c-Myb activity. We also report that PMCA1 expression is regulated during VSMC cell cycle progression, such that levels of PMCA1 are 40% lower at the G1/S interface than at G0. Moreover, transient overexpression of PMCA1a in VSMC elevates the 45Ca efflux rate by approximately 2-fold, decreases resting and peak thapsigargin-releasable Ca2+ concentrations at G1/S by 43% (68 nM) and 52% (160 nM), respectively, and reduces the rate of cell proliferation by over 2.5-fold. These data define a mechanism for c-Myb-dependent Ca2+ homeostasis and support a critical role for PMCA in the regulation of VSMC growth. PMID- 9227425 TI - F-actin disruption attenuates agonist-induced [Ca2+], myosin phosphorylation, and force in smooth muscle. AB - Cytochalasins B and D (at 10 microM) inhibited stress development induced by 1 microM carbachol in bovine tracheal smooth muscle by 55% and 90%, respectively. Glucose depletion was ineffective in inhibiting carbachol-induced contraction, indicating that inhibition of glucose transport was not the cause. Cytochalasin D treated smooth muscle cells appeared collapsed, with spiky protrusions from the cell membrane. Deconvolution of fluorescent images of fluorescein isothiocyanate phalloidin-labeled smooth muscle cells revealed concentrations of actin filaments near the cell periphery, including near the spiky protrusions. Cytochalasin B attenuated carbachol-induced intracellular Ca2+ concentration ([Ca2+]), especially the initial peak intracellular [Ca2+]. Cytochalasin B also attenuated carbachol-induced myosin light chain phosphorylation. However, when the myosin phosphorylation data were plotted against time-matched intracellular [Ca2+] data, the two relationships in control and cytochalasin B-treated smooth muscle were similar, suggesting that the changes in myosin phosphorylation could be explained by the changes in intracellular [Ca2+]. These results suggest that actin filaments in smooth muscle cells are dynamic and may be an integral component of Ca2+ regulation and/or signal transduction in receptor-coupled mechanisms. PMID- 9227426 TI - Shrinkage-induced activation of Na+/H+ exchange: role of cell density and myosin light chain phosphorylation. AB - Previously, we suggested that myosin light chain kinase (MLCK) is involved in shrinkage-induced activation of the Na+/H+ exchanger in rat astrocytes. Here we have studied the effects of hyperosmotic exposure in C6 glioma cells, a common model for astrocytes. Shrinkage-induced activation of the Na+/H+ exchanger in C6 cells is directly proportional to the degree of shrinkage, results in an alkaline shift in the pK' of the exchanger, is dependent on ATP, and is inhibited by ML-7 (an MLCK inhibitor) and by various calmodulin inhibitors. Cell shrinkage also results in increased phosphorylation of myosin light chain (MLC). Interestingly, shrinkage-induced activation of the exchanger does not occur in subconfluent C6 cells. However, phosphorylation of MLC still occurs in subconfluent cultures of C6 cells on shrinkage, suggesting that the lack of activation in these cells occurs at a point between MLC phosphorylation and Na+/H+ exchange activation. The lack of activation of Na+/H+ exchange in subconfluent C6 cells can be utilized to further elucidate the shrinkage-induced activation pathway. PMID- 9227428 TI - Survival of human epidermal keratinocytes after short-duration high temperature: synthesis of HSP70 and IL-8. AB - Thermal injury by short pulses (1-30 s) of relatively high temperature (50-68 degrees C) was investigated in normal human epidermal keratinocytes (NHEK). NHEK were cultured on plastic cover-slips and dipped in medium held at various temperatures. Survival assessed by methylthiazol tetrazolium reduction assay at 6 days postheating demonstrated an inverse time-temperature relationship that indicated that most cells could survive after a 1-s, 60 degrees C exposure or a 30-s, 55 degrees C exposure. Arrhenius plots of the data indicated major transition points for cell injury at 50 and 60 degrees C. Heat shock protein 70 (HSP70) and interleukin-8 (IL-8) were both induced by elevation of temperature between 50 and 60 degrees C for as short a time as 1 s. HSP70 synthesis stimulated by short, high pulses of heat appeared to induce thermotolerance. These results demonstrate that brief exposure to relatively high temperature can induce HSP70 and IL-8 synthesis in keratinocytes. PMID- 9227427 TI - Confocal imaging analysis of ATP-induced Ca2+ response in individual endothelial cells of the artery in situ. AB - The mechanisms for mobilization of intracellular free Ca2+ have been studied in various types of isolated and cultured cells, but little is known about Ca2+ mobilization in individual cells in situ. We tried to establish imaging analysis of intracellular free Ca2+ concentration ([Ca2+]i) in individual cells loaded with the acetoxymethyl ester of fluo 3 in situ, using laser scanning confocal microscopy. The method permitted us to distinguish signals from endothelial and smooth muscle cells of guinea pig artery. Addition of ATP to the artery caused a transient increase in endothelial [Ca2+]i. It was concluded that the response was induced via P2Y purinoceptors, because adenosine 5'-O-(2-thiodiphosphate), but not UTP, caused a similar response independent of extracellular Ca2+. The percentage of cells that responded to ATP (1-10 microM) and the peak amplitude of the transient increase in [Ca2+]i were dose dependently increased. Using rapid xy scanning and line-scanning modes, we confirmed that 10 microM ATP induced Ca2+ waves, at a rate of 10-30 microns/s, after a lag time of approximately 3 s. These results show that [Ca2+]i waves within endothelial cells are physiologically induced by ATP via P2Y purinoceptor, but not P2U purinoceptor, in aortic strips in situ. The method should be of use in the study of vascular physiology and pathophysiology. PMID- 9227429 TI - Differential processing of guanylyl cyclase C along villus-crypt axis of rat small intestine. AB - Many strains of enterotoxigenic Escherichia coli produce a heat-stable peptide enterotoxin (STa) that binds to the intestinal receptor guanylyl cyclase C (GC C). STa receptors are structurally heterogeneous, but the molecular events causing this heterogeneity remain obscure. We examined the influence of cell position along the villus-crypt axis on STa receptor heterogeneity by fractionating EDTA-dissociated cells that detached in a villus-to-crypt direction. STa affinity labeling experiments revealed that the initially released villus "tip" fraction had four major STa binding proteins (STBPs), with relative molecular weight (M(r)) of 150,000, 135,000, 125,000, and 95,000, that did not react with a GC-C carboxy-terminal antibody. Yet succeeding villus cell fractions had major immunoreactive STBPs with M(r) of 275,000 and 250,000. Limited proteolysis of these larger GC-C isoforms produced 1) smaller STBPs that had M(r) similar to those in the initial villus fraction, 2) a 65,000 M(r) protein GC-C isoform that did not bind STa, and 3) elevated basal and STa-induced cyclase activity. Our data show that STBP structural heterogeneity in the intact intestine arises largely from multisite proteolytic processing of GC-C. PMID- 9227430 TI - cAMP and forskolin inhibit potassium currents in rat taste receptor cells by different mechanisms. AB - In gustatory transduction, adenosine 3',5'-cyclic monophosphate (cAMP) has been suggested to close potassium channels when elevated by sweet stimuli or to open cAMP-gated cation channels when depressed by bitter stimuli. These experiments examine the effect of cAMP on whole cell currents from posterior taste receptor cells with standard patch-clamp techniques. Elevating cytosolic cAMP by pipette administration, membrane-permeant analogs [8-(4-chlorophenylthio)-cAMP (CPT-cAMP) and dibutyryl-cAMP], or by phosphodiesterase inhibition [3-isobutyl-1 methylxanthine (IBMX)] produced poorly reversible inhibitions of outward potassium currents by up to 33%. Unexpectedly, middle to high concentrations of forskolin (> 5 microM) profoundly and reversibly inhibited these currents (95%) with greatly accelerated inactivation kinetics. 1,9-Dideoxyforskolin, an ineffective activator of adenylate cyclase, was similarly potent. Kinase inhibitors effectively blocked the effects of cAMP elevations produced by IBMX or CPT-cAMP but did not block these forskolin actions. However, at low concentrations (5 microM), forskolin reduced potassium currents in a phosphorylation-dependent manner. Collectively, these data suggest that cAMP produces a phosphorylation-dependent inhibition of outward potassium currents but that forskolin's actions are independent of cAMP or phosphorylation except at low concentration. cAMP was also effective in altering the waveform of the gustatory action potential, implying it may modify transmission of gustatory information to the brain. PMID- 9227431 TI - G protein-coupled receptors control vascular smooth muscle cell proliferation via pp60c-src and p21ras. AB - The binding of vasoactive peptides to their respective G protein-coupled receptors has been implicated in the pathogenesis of vascular smooth muscle cell proliferation, leading to the development of hypertension, arteriosclerosis, and restenosis after vascular injury. We previously showed that the cytosolic tyrosine kinase pp60c-src is crucial for angiotensin II (ANG II)-induced activation of the protooncogene p21ras. Therefore, we investigated the role of pp60c-src and p21ras in rat aortic smooth muscle cell proliferation induced by several G protein-coupled receptors. ANG II, endothelin-1, or thrombin increased cell proliferation and DNA synthesis. Electroporation of anti-pp60c-src antibodies into cells abolished proliferation in response to these G protein coupled receptor ligands but not in response to platelet-derived growth factor-BB (PDGF-BB). In contrast, electroporation of anti-p21ras antibody completely blocked DNA synthesis and cell proliferation in response to ANG II, endothelin-1, thrombin, and PDGF-BB. Our data indicate that the pp60c-src tyrosine kinase is necessary and specific for vascular smooth muscle cell proliferation and DNA synthesis in response to G protein-coupled receptors but not classic growth factor receptors. PMID- 9227432 TI - Regulation of branched-chain ketoacid dehydrogenase flux by extracellular pH and glucocorticoids. AB - In muscles of rats with metabolic acidosis, branched-chain alpha-ketoacid dehydrogenase (BCKAD) activity is increased. Potential stimulatory signals include acidemia and/or glucocorticoids. It is unclear whether the signal(s) increases BCKAD activity by changing the activation state of the enzyme or by increasing the amount of enzyme. To separate the influences of extracellular pH and glucocorticoids on leucine catabolism, maximal BCKAD flux and the activation state (the ratio of basal to total flux) were measured in two cell types: 1) cells that do not express glucocorticoid receptors and 2) cells stably transfected to express glucocorticoid receptors. Acidification (pH 6.95) increased 1) the activation state from 67.2% at pH 7.4 to 82.8% at pH 6.95, 2) maximal BCKAD flux by 50%, and 3) the BCKAD subunit contents in both cell types (57, 410, and 270% for E2, E1 alpha, and E1 beta, respectively). Dexamethasone increased the BCKAD activation state from 67.2 to 82.3% in cells expressing glucocorticoid receptors, whereas dexamethasone plus acidification increased the activation state to 98%. The time course of stimulation by dexamethasone was slower than that by acidification. These results demonstrate that BCKAD is differentially regulated by extracellular pH and glucocorticoids. PMID- 9227433 TI - Modeling enrichment kinetics from dynamic 13C-NMR spectra: theoretical analysis and practical considerations. AB - Measurements of oxidative metabolism in the heart from dynamic 13C nuclear magnetic resonance (NMR) spectroscopy rely on 13C turnover in the NMR-detectable glutamate pool. A kinetic model was developed for the analysis of isotope turnover to determine tricarboxylic acid cycle flux (VTCA) and the interconversion rate between alpha-ketoglutarate and glutamate (F1) by fitting the model to NMR data of glutamate enrichment. The results of data fitting are highly reproducible when the noise level is within 10%, making this model applicable to single or grouped experiments. The values for VTCA and F1 were unchanged whether obtained from least-squares fitting of the model to mean experimental enrichment data with standard deviations in the cost function (VTCA = 10.52 mumol.min-1.g dry wt-1, F1 = 10.67 mumol.min-1.g dry wt-1) or to the individual enrichment values for each heart with the NMR noise level in the cost function (VTCA = 10.67 mumol.min-1.g dry wt-1, F1 = 10.18 mumol.min-1.g dry wt 1). Computer simulation and theoretical analysis indicate that glutamate enrichment kinetics are insensitive to the fractional enrichment of acetyl-CoA and changes in small intermediate pools (< 1 mumol/g dry wt). Therefore, high resolution NMR analysis of tissue extracts and biochemical assays for intermediates at low concentrations are unnecessary. However, a high correlation between VTCA and F1 exists, as anticipated from competition for alpha ketoglutarate, which indicates the utility of introducing independent experimental constraints into the data fitting for accurate quantification. PMID- 9227434 TI - TCA cycle flux estimates from NMR- and GC-MS-determined [13C]glutamate isotopomers in liver. AB - Infusion of 13C-labeled lactate into rabbits and the subsequent measurement of glutamate isotopomers by 13C nuclear magnetic resonance (NMR) spectroscopy enables one to calculate relative flow rates associated with the tricarboxylic acid (TCA) cycle, albeit with a lower precision than one would obtain using a perfused organ. Two factors contribute to the lower precision in the determination of relative flow rates for the in vivo system: 1) a poorly defined pyruvate input and 2) low levels of 13C-enriched oxaloacetate and acetyl-CoA isotopomers, which give rise to weaker glutamate isotopomer NMR signals. To help overcome these limitations, we introduce a procedure to 1) include experimental data from gas chromatography-mass spectrometry (GC-MS) and 2) account for the uncertainty in the labeling of the input to pyruvate by creating the labeling as a measurement that is subject to measurement error. The effects of the uncertainties in the input labeling, NMR data, and MS data are evaluated via a Monte Carlo method. The change in the precision of the relative fluxes for the cases of high/low NMR and high/low MS precision is given. An uncertainty in the lactate measurements of up to 10% does not add significantly to the imprecision of the relative flow rates. PMID- 9227436 TI - Quantitative analysis of acetoacetate metabolism in AS-30D hepatoma cells with 13C and 14C isotopic techniques. AB - Experimental hepatoma cells utilize acetoacetate as an oxidative energy source and as a precursor for lipid synthesis. The significance of ketone body metabolism in tumors lies in the study of tumor-host metabolism and the ketoneMic condition that is often present in cancer patients. The quantitative importance of acetoacetate and glucose was investigated in AS-30D cells with use of 13C and 14C isotopic methods. In addition, the effects of acetoacetate were compared with those of dichloroacetic acid (DCA), an activator of pyruvate dehydrogenase (PDH). The 14CO2 ratio method evaluated the entry of pyruvate into the tricarboxylic acid (TCA) cycle and revealed that acetoacetate diverted pyruvate from PDH to pyruvate carboxylation. In contrast, DCA increased the oxidation of glucose largely through PDH, indicating that PDH is not maximally active in the absence of DCA. Isotopomer spectral analysis of lipid synthesis demonstrated that, in the absence of acetoacetate, glucose supplied 65% of the acetyl-CoA used for de novo lipogenesis. When 5 mM acetoacetate was included in the incubation, glucose was displaced as a lipogenic precursor and acetoacetate supplied 85% of the acetyl CoA for lipogenesis vs. only 2% for glucose. Thus AS-30D cells have a large capacity for acetoacetate utilization for de novo lipogenesis. PMID- 9227435 TI - Contractile properties of diaphragm muscle segments from old mdx and old transgenic mdx mice. AB - Diaphragm muscles of young (4- to 6-mo-old) mdx mice show severe fiber necrosis and have normalized forces and powers 60 and 46% of the values for control C57BL/10 mice. In contrast, microinjection of mdx mouse embryos with a truncated dystrophin minigene has produced young transgenic mdx (tg-mdx) mice with a level of dystrophin expression and structural and functional properties of diaphragm muscle strips measured in vitro not different from those of control mice. Whether dystrophin expression and functional corrections persist for the life span of these animals is not know. We tested the null hypothesis that, in old (24 mo) tg mdx mice, dystrophin expression is adequate and diaphragm muscle strips have forces and powers not different from values for diaphragm muscle strips from young tg-mdx mice or control mice. Compared with control values, diaphragm muscle strips from old mdx mice had normalized forces and powers of 48 and 31%, respectively. Expression of dystrophin persisted in diaphragm muscles of old tg mdx mice, and functional properties were not different from diaphragm muscles of young tg-mdx or young or old control mice. These results suggest that, with a transgenic animal approach, dystrophin expression and functional corrections persist for the life span of the animals. PMID- 9227437 TI - Effects of endotoxin on zinc metabolism in human volunteers. AB - After stress or trauma, the serum zinc concentration decreases. This study evaluated possible mechanisms for hypozincemia with the use of a human endotoxemia model. Two doses of endotoxin [lipopolysaccharide (LPS)] were administered on consecutive mornings to 12 healthy volunteers, and each subject was also studied after saline injection. Blood was analyzed for zinc, cytokines (tumor necrosis factor-alpha and interleukin-6), albumin, albumin-zinc binding, and C-reactive protein (CRP). Serial 24-h urine collections were analyzed for zinc. Each LPS dose briefly increased plasma cytokine concentrations and decreased the serum zinc concentration. Serum albumin, the major zinc binding protein, did not decrease, but a progressive increase in CRP was found. LPS did not alter zinc binding affinity to serum albumin. Urine zinc losses were not increased. We conclude that hypozincemia in this model cannot be explained by decreased serum albumin, changes in serum albumin-zinc binding, or increased urinary zinc excretion. Because hypozincemia was transient and followed cytokine peaks, we postulate that LPS-stimulated hypozincemia is mediated, at least partly, by a cytokine-directed internal redistribution of zinc. PMID- 9227438 TI - Lactate production by swine adipocytes: effects of age, nutritional status, glucose concentration, and insulin. AB - To develop an alternative model in which to study the relationship between adipose tissue lactate production, obesity, and non-insulin-dependent diabetes mellitus (NIDDM), we investigated lactate production by swine adipocytes. Subcutaneous adipocytes from fasted 3-wk-old, fasted 7-mo-old, and fed 7-mo-old Yucatan minIature swine were isolated and incubated with 0.2, 1, 5, 10, or 25 mM glucose +/- 1 mU/ml insulin. Total glucose metabolism (TGM) was estimated by product summation. Results showed that 1) TGM was threefold greater in cells from fasted 7-mo- vs. 3-wk-old swine (P < 0.05), 2) TGM was 2.7-fold greater in cells from fed 7-mo-old vs. fasted 7-mo-old swine (P < 0.05), 3) insulin failed to stimulate TGM in adipocytes from swine of either age and either nutritional status, and 4) lactate and pyruvate accounted for 34 and 30% of TGM, respectively, in adipocytes from swine of both ages. Similarities in glucose metabolism and lactate production in adipocytes from swine and obese NIDDM humans make the swine a potentially valuable model for studying lactate production associated with obesity and NIDDM. PMID- 9227439 TI - Osteogenic protein-1 downregulates endothelin A receptors in primary rat osteoblasts. AB - Osteogenesis is a complex process whereby growth factors and mediators from both local and systemic sources modulate the bone-forming activities of osteoblasts. In the present study we utilized primary cultures of fetal rat calvarial cells to characterize osteoblast responsiveness to the vascular mediator endothelin-1 (ET 1) and to investigate whether ET-1 responses are regulated by osteogenic protein 1 (OP-1). We found that a 1- to 2-day exposure to OP-1 diminished ET-1 receptor ligand binding and signal transduction by downregulating ET-1 receptor mRNA expression. ET-1-mediated calcium signaling and ligand binding were completely abolished by the ETA receptor antagonist BQ-123, suggesting that ET-1 effects are mediated by this receptor. Northern analysis of total RNA revealed that ETA mRNA expression was inhibited approximately 50% by OP-1 treatment, whereas ETB receptor mRNA was not detected by this method of analysis. In OP-1-treated cultures, the magnitude and duration of ET-1 calcium signals varied among individual cells. This finding may be related to a heterogeneous OP-1 response, indicated by alkaline phosphatase induction in only a subpopulation of cells. These results suggest that modulation of osteoblast function by ET-1 occurs during distinct periods of phenotypic development and imply that downregulation of ET-1 responsiveness may be necessary for optimal bone formation in vivo. PMID- 9227440 TI - Age-related increase in muscle sympathetic nerve activity is associated with abdominal adiposity. AB - Tonic sympathetic nerve activity (SNA) increases with age, but the mechanisms are unknown. There is evidence that SNA is positively related to total and abdominal body fat, which also increase with age. We tested the hypotheses that 1) the elevation in SNA with age is partially accounted for by higher abdominal and/or total body fat and 2) skeletal muscle is a target of the adiposity-related sympathetic effects. Direct microneurographic recordings of skeletal muscle SNA (MSNA) were obtained during supine rest in 16 older (64 +/- 1 yr, means +/- SE) and 16 young (24 +/- 1 yr) adult males. Central body fat was estimated by waist circumference (WC) and fat mass (FM) by hydrostatic weight. MSNA, WC, and FM were higher in the older vs. young males (44 +/- 2 vs. 22 +/- 2 bursts/min, 91 +/- 2 vs. 79 +/- 1 cm, and 19 +/- 2 vs. 9 +/- 1 kg, respectively; all P < 0.0001). Although univariate correlations were high for MSNA and both WC (r = 0.77) and FM (r = 0.75), stepwise multiple regression analysis revealed WC to be the best predictor of MSNA (R2 = 0.60, P < 0.0001), with FM explaining only an additional 2% of the variance (not significant). Statistically covarying for WC reduced but did not eliminate the difference in adjusted age-group means for MSNA (39 +/- 3 vs. 26 +/- 2 bursts/min, P = 0.003). We conclude that 1) the elevated SNA in older adults is partially related to higher body fat, particularly in the abdominal region, and 2) skeletal muscle is a target of the adiposity-related sympathetic effects observed with aging. PMID- 9227441 TI - Dynamic aspects for interislet synchronization of oscillatory insulin secretions. AB - How are the oscillatory insulin secretions from numerous islets synchronized to result in an identifiable oscillation? We postulated that a sudden increase in glucose concentration could best account for the interislet synchronization. The perifusion with two parallel chambers each containing 100 islets from the same rat was performed. The glucose concentrations of two chambers were simultaneously increased from 100 to 300 mg/dl in step function to examine the synchronizing efficacy. Synchrony and regularity of insulin oscillation were evaluated by cross correlation and/or power spectral analysis. Although the insulin had been in stable oscillation, we found that the synchrony between two chambers and the regularity of each chamber were still significantly improved after a sudden increase in glucose level. However, the improved synchrony and regularity were transient. They gradually slid toward a less rigorous condition in a 15-h long term perifusion. We suggested that the interislet synchronization of oscillatory insulin secretions could be improved by a sudden increase in glucose level. The insulin pulses were therefore enhanced to present their physiological effects. PMID- 9227442 TI - Antihypertensive and vasculo- and renoprotective effects of pioglitazone in genetically obese diabetic rats. AB - Although an improvement of insulin sensitivity has been shown to be a new therapeutic approach for treating diabetes mellitus, details of effects of this treatment on the cardiovascular system and possible renal complications remain unknown. In the present study, we investigated the effects of a thiazolidine derivative, pioglitazone, and examined the insulin-sensitizing action on blood pressure, nephropathy, and vascular changes in genetically obese diabetic Wistar fatty (WF) rats. Pioglitazone (3 mg.kg-1.day-1) was orally administered for 13 wk starting at the age of 5 wk, and the results were compared with those of vehicle treated WF rats. At the age of 18 wk, vehicle-treated WF rats were associated with mild hypertension, nephropathy with proteinuria histological glomerular injury, and renal arteriolosclerosis in addition to hyperglycemia, hyperinsulinemia, and hyperlipidemia. Treatment with pioglitazone significantly improved glucose and lipid metabolism. In addition, it lowered blood pressure, decreased proteinuria, and prevented glomerular injury, renal arteriolosclerosis, and aortic medial wall thickening, whereas body weight, food intake, sodium balance, and urinary norepinephrine excretion were significantly increased. These results suggest that the insulin-sensitizing agent pioglitazone is effective in correcting not only glucose and lipid metabolism but also cardiovascular and renal complications in non-insulin-dependent diabetes mellitus. PMID- 9227443 TI - Effect of short-term propionate infusion on feed intake and blood parameters in sheep. AB - The hypothesis that propionate is a short-term feed intake-regulating agent was studied. Mature wether sheep were infused over 20 min with Na propionate into the mesenteric vein, while feed intake and feeding pattern were monitored over 1.5 h. Feed intake was reduced by infusions at 2 mmol/min, which were associated with marked increases in jugular as well as portal concentrations of insulin, glucose, and propionate. In a second experiment, animals were infused with 2 mmol/min Na propionate into the portal vein. No decrease in feed intake was observed, although there were similar increases in insulin, glucose, and propionate as found in mesenteric vein-infused animals. It is concluded that mesenteric propionate in high doses acts as a satiety factor. Possible explanations for the difference between site of infusion may be a different distribution of the infusate over the liver and/or the presence of propionate-sensitive receptors in the mesenteric/portal vein region. It seems unlikely that insulin concentrations are involved in inducing satiety in propionate-infused animals. PMID- 9227444 TI - Changes in cytokine production and T cell subpopulations in experimentally induced zinc-deficient humans. AB - We have utilized an experimental model of human zinc deficiency for study of cytokines production by TH1 and TH2 cells. Additionally, we determined ratios of CD4+ to CD8+ and CD4+ CD45RA+ to CD4+CD45RO+ cells and percentages of CD73+ T cytolytic cells in the CD8+ subset. The data were collected during baseline, at the end of the zinc-restricted period, and following zinc repletion. Our results showed that functions of TH1 cells, as evidenced by production of interferon gamma, interleukin-2 (IL-2), and tumor necrosis factor-alpha, were decreased, whereas functions of TH2 cells (production of IL-4, IL-6, and IL-10) were unaffected by zinc deficiency. Thus an imbalance between TH1 and TH2 cells resulted because of zinc deficiency in humans. Our studies also showed that zinc may be required for regeneration of new CD4+ T lymphocytes and maintenance of T cytolytic cells. We conclude that an imbalance between TH1 and TH2 cells, decreased recruitment of T naive cells, and decreased percentage of T cytolytic cells may account for decreased cell-mediated immune functions in zinc-deficient subjects. PMID- 9227445 TI - Increased expression of nitric oxide synthase in the myometrium of the pregnant rat uterus. AB - Nitric oxide (NO) relaxes uterine smooth muscle and is produced by the pregnant uterus. Our previous studies revealed an increase in rat uterine NO synthase (NOS) activity in pregnancy and a decline at term. In the present study, we have examined the distribution of NOS isoform expression to determine whether their regulation is consistent with a role in the inhibition of uterine contractions before term. At day 17-18 of pregnancy, NOS immunohistochemistry revealed expression of two isoforms: endothelial constitutive form of NOS (ecNOS) in vascular endothelium and inducible form of NOS (iNOS) in myometrial and vascular smooth muscle and in decidual epithelium. Immunoblotting revealed that expression of iNOS declined nearly fivefold, whereas ecNOS declined twofold in laboring rats at term. We conclude that iNOS is expressed in myometrium of pregnant rat uterus but not the virgin rat and that iNOS expression declines at term when labor is present. The pattern of changes in myometrial iNOS expression with advancing gestation suggests that NO could act in an autocrine and/or paracrine manner to inhibit uterine contractions before term. PMID- 9227446 TI - 17 beta-Estradiol modulation of glucose transporter 1 expression in blood-brain barrier. AB - The present study was designed to evaluate 17 beta-estradiol (E2) modulation of glucose transporter 1 (GLUT-1) protein and mRNA expression in blood-brain barrier (BBB) endothelium. Female rats were ovariectomized (OVX) for 12-14 days, then E2 was injected at dosages of 1-100 micrograms/kg sc at 2-16 h before sampling. Glucose transport into BBB endothelial cells was assessed using 2-deoxy [14C]glucose (2-[14C]DG) uptake. GLUT-1 protein and mRNA samples were analyzed by Western and Northern blotting, respectively. E2 treatment caused dose- and time dependent increases in 2-[14C]DG uptake and GLUT-1 protein expression by microvessels. The peak responses were induced by 10 micrograms/kg E2 dose at the 4-h sampling time (36.0 and 31.3% increases, P < 0.05, respectively). GLUT-1 mRNA demonstrated a transient increase at 15 min (55%, P < 0.05), then decreased to basal level by 2 h. This study shows that in vivo treatment with E2 increases 2 [14C]DG uptake into the BBB endothelial cells and suggests this E2 effect is due to its modulation of GLUT-1 mRNA and protein. PMID- 9227447 TI - Effect of amino acid and glucose administration during postexercise recovery on protein kinetics in dogs. AB - To examine the effect of the timing of amino acids (AA) and glucose (G) administration after exercise on protein kinetics, ten dogs fitted with chronic catheters in the artery and the femoral vein ran on a treadmill for 150 min. They were intraportally infused with a solution containing AA and G either right after (E) or 2 h after (L) the exercise. The protein kinetics were estimated using the arteriovenous difference of phenylalanine (Phe) coupled with the [2H5]Phe dilution method. The net balance of Phe across the hindlimb (HL) was negative after exercise. It became positive in E within 15 min after the start of the infusion, and it remained negative in L until the infusion was initiated. The uptake of Phe by the HL during the second half of the infusion period was higher in E than in L (10.9 +/- 6.6 vs. 5.4 +/- 2.3 nmol.kg-1.min-1, P = 0.049). During the infusion, protein synthesis in the HL was higher in E than in L (29.7 +/- 9.6 vs. 22.0 +/- 10.1 nmol.kg-1.min-1, P = 0.028), whereas proteolysis was comparable (18.7 +/- 5.7 vs. 16.5 +/- 11.1 nmol.kg-1.min-1. These results suggest that the early provision of the nutrients after exercise more effectively enhances protein accretion than nutrients administered later. PMID- 9227449 TI - Effects of leucine on whole body leucine, valine, and threonine metabolism in humans. AB - We tested whether expansion of the plasma leucine pool distorts leucine or valine tracer kinetics, causing errors in the derived values of whole body proteolysis. Seven normal adults received a 10-h primed-continuous tracer infusion of L-[5,5,5 2H3]leucine, L-[(1-13)C]valine, and L-[(1-13)C]threonine, during the final 7 h of which L-leucine was infused at a rate that more than tripled the plasma leucine concentration. Leucine, valine, and threonine rates of appearance were converted to a common value of whole body proteolysis on the basis of their concentrations in body proteins. The conversion of labeled leucine and valine to their corresponding branched-chain alpha-keto and alpha-hydroxy acids was also monitored. Before the unlabeled leucine infusion, postabsorptive whole body proteolysis was estimated similarly by the three tracers (approximately 180 mg protein.kg-1.h-1. The leucine infusion reduced proteolysis by an average of 21% (P < 0.006), as estimated by use of valine or threonine kinetics, and by 10% by use of leucine kinetics (P < 0.02). No delay in the conversion of valine to alpha ketoisovalerate occurred during the leucine infusion. Thus all three tracers indicated similar postabsorptive rates of whole body proteolysis and a reduction of proteolysis during leucine administration, although the magnitude of the effect was underestimated with use of the leucine tracer. PMID- 9227448 TI - beta 3-Adrenergic-mediated suppression of leptin gene expression in rats. AB - To investigate the role of beta 3-adrenergic receptors in the suppression of leptin gene expression, we fasted F-344 rats to decrease leptin mRNA levels, refed the rats to stimulate leptin mRNA production, and examined the ability of the beta 3-adrenergic agonist CGP-12177 to prevent the rise in leptin mRNA levels. In the initial 2 h after CGP-12177 (0.75 mg/kg), there were significant reductions in both food consumption and leptin mRNA levels in epididymal, perirenal, and interscapular white adipose tissue. We were unable to detect leptin mRNA in interscapular brown adipose tissue (IBAT), whereas there was a significant increase in uncoupling protein mRNA levels in IBAT after CGP-12177. The suppression of leptin mRNA and food intake by CGP-12177 was confirmed in a second experiment using another rat strain, the F-344 x BN. Furthermore, refeeding after a period of fasting increased leptin mRNA, which was prevented by CGP-12177. These data indicate a role for beta 3-adrenergic-mediated regulation of leptin gene expression in nonmutant rodents and are consistent with other reports suggesting that beta 3-adrenergic agonists suppress food intake. PMID- 9227450 TI - Adenylyl cyclase inhibitory pathway is differentially modified in rat white and brown fat by high-energy diets. AB - Incubation of white adipose tissue (WAT) adipocytes from rats fed a high-energy diet (Exp group) with antilipolytic Gi-coupled adenylyl cyclase inhibitory agonists, nicotinic acid (Nic) and N8-(L-2-phenylisopropyl)adenosine (PIA), resulted in lower cellular adenosine 3',5'-cyclic monophosphate (cAMP) levels than in stimulated adipocytes from rats fed a nutritionally balanced diet (Con group). In contrast to WAT, incubation of brown adipose tissue (BAT) adipocytes with Nic yielded higher cAMP levels in the Exp vs. Con rats. In both WAT and BAT adipocytes, pertussis toxin treatment abolished the differences in Nic- and PIA inhibited cAMP formation between Exp and Con animals. Immunoblotting of adipocyte membranes indicated a lower content of Gi alpha but not Gs alpha in BAT membranes of Exp vs. Con animals after 6 and 10 wk of feeding. No such differences were found in the Gs alpha or Gi alpha contents of WAT membranes. Thus the inhibitory pathway of adenylyl cyclase is proposed to be sensitized in WAT and desensitized in BAT of rats fed high-energy diets. These modifications in sensitivity are in line with reduced cAMP and lipolysis in WAT and increased cAMP and thermogenesis in BAT during obesity. PMID- 9227451 TI - Intestinal function in mice with small bowel growth induced by glucagon-like peptide-2. AB - Glucagon-like peptide-2 (GLP-2) stimulates small intestinal growth through induction of intestinal epithelial proliferation. To examine the physiology of GLP-2-induced bowel, mice were treated with GLP-2 (2.5 micrograms) or vehicle for 10 days. Small intestinal weight increased to 136 +/- 2% of controls in GLP-2 treated mice, in parallel with 1.4 +/- 0.1- and 1.9 +/- 0.5-fold increments in duodenal RNA and protein content, respectively (P < 0.05-0.001). Similarly, the activities of duodenal maltase, sucrase, lactase, glutamyl transpeptidase, and dipeptidyl-peptidase IV (215 +/- 28% of controls; P < 0.001) were increased by GLP-2. Oral or duodenal administration of glucose or maltose did not reveal any differences in the ability of GLP-2-treated mice to absorb these nutrients, possibly because of decreases in expression of the glucose transporters sodium dependent glucose transporter-1 (SGLT-1) and GLUT-2. In contrast, absorption of leucine plus triolein was increased after duodenal administration in GLP-2 treated mice (P < 0.01-0.001). Finally, GLP-2 did not alter other markers of intestinal or pancreatic gene expression, including levels of mRNA transcripts for ornithine decarboxylase, multidrug resistance gene, amylase, proglucagon, proinsulin, and prosomatostatin. Thus induction of intestinal growth by GLP-2 in wild-type mice results in a normal-to-increased capacity for nutrient digestion and absorption in vivo. PMID- 9227452 TI - Glucose production and gluconeogenesis in adults with uncomplicated falciparum malaria. AB - Although glucose production is increased in severe malaria, the influence of uncomplicated malaria on glucose production is unknown. Therefore, we measured in eight adult Vietnamese patients with uncomplicated falciparum malaria and eight healthy Vietnamese controls glucose production (by infusion of [6,6-2H2]glucose) and the fractional contribution of gluconeogenesis (by oral ingestion of 2H2O); glycogenolysis was calculated as the difference between the two. After 20 h of fasting, plasma glucose was 4.7 +/- 0.2 mmol/l in the patients and 4.3 +/- 0.2 mmol/l in the controls (not significant). Glucose production was approximately 25% higher in the patients (16.9 +/- 1.3 vs. 13.4 +/- 0.3 mumol.kg-1.min-1, P = 0.01). Fractional and absolute gluconeogenesis were increased in the patients (approximately 87 vs. approximately 59%, P < 0.001; and 14.6 +/- 1.3 vs. 7.9 +/- 0.2 mumol.kg-1.min-1, P < 0.001, respectively). The contribution of glycogenolysis to total glucose production was decreased in the patients: 2.3 +/- 0.5 vs. 5.5 +/- 0.4 mumol.kg-1.min-1 (P < 0.002). In conclusion, in adult patients with uncomplicated falciparum malaria, glucose production is increased by approximately 25% due to an increased rate of gluconeogenesis, whereas glycogenolysis is decreased. The mechanism by which these changes occur is uncertain. However, counterregulatory hormone and cytokine concentrations were increased in the patients. PMID- 9227453 TI - Regulation of plasma fatty acid oxidation during low- and high-intensity exercise. AB - In the present study we examined the hypothesis that fatty acid oxidation is less during high-intensity exercise than during moderate-intensity exercise because of inhibition of long-chain fatty acid entry into the mitochondria. Six volunteers exercised at 40% peak oxygen consumption (VO2peak) for 60 min and at 80% VO2peak for 30 min on two different occasions. [1-13C]oleate, a long-chain fatty acid, and [1-14C]octanoate, a medium-chain fatty acid, were infused for the duration of the studies. Lipids and heparin were infused during exercise at 80% VO2peak to prevent the expected decrease in plasma free fatty acid (FFA) concentration. Plasma oleate and total FFA availability were similar in the two experiments. Oleate oxidation decreased from 2.8 +/- 0.6 (40% VO2peak) to 1.8 +/- 0.2 mumol.kg 1.min-1 (80% VO2peak, P < 0.05), whereas octanoate oxidation increased from 1.0e 05 +/- 1.0e-06 (40% VO2peak) to 1.3e-05 +/- 5.1e-06 mumol.kg-1.min-1 (80% VO2peak, P < 0.05). Furthermore, the percentage of oleate uptake oxidized decreased from 67.7 +/- 2.8% (40% VO2peak) to 51.8 +/- 4.6% (80% VO2peak, P < 0.05), whereas the percentage of octanoate oxidized was similar during exercise at 40 and 80% VO2peak (84.8 +/- 2.7 vs. 89.3 +/- 2.7%, respectively). Our data suggest that, in addition to suboptimal FFA availability, fatty acid oxidation is likely limited during high-intensity exercise because of direct inhibition of long-chain fatty acid entry into mitochondria. PMID- 9227454 TI - Growth hormone-induced insulin resistance: role of the insulin receptor, IRS-1, GLUT-1, and GLUT-4. AB - Treatment of rats with growth hormone (GH; 1 mg/kg sc) twice daily over 2.5 days did not alter fasting plasma glucose or glucose tolerance but increased fasting plasma insulin levels 65% and peak insulin response to a glucose load 35% over controls, indicating the development of insulin resistance. Studies on partially purified insulin receptors from soleus muscles showed that GH increased the abundance of insulin receptor beta-subunits by 48% as measured by immunoblotting. Despite this increase, GH abolished the increase in autophosphorylation of the insulin receptor beta-subunit in response to physiological hyperinsulinemia and diminished by 28% the response to supraphysiological hyperinsulinemia. Similarly, insulin-stimulated phosphorylation of insulin receptor substrate-1 (IRS-1) was decreased 25% by GH, but the abundance of IRS-1 was not affected. Studies on rats pretreated with streptozotocin suggested that the effects of GH are direct and not secondary to GH-induced hyperinsulinemia. GH decreased basal GLUT-1 abundance in the low-density microsome and plasma membrane fractions of epididymal adipocytes by 50 and 42%, respectively, but decreased basal GLUT-4 abundance only in the low-density microsome fraction by 24%. Despite these alterations, the abundance of both transporters in the plasma membrane fraction of adipocytes incubated with 0.1 U insulin/ml was not diminished by GH. PMID- 9227455 TI - Increased activity of the hexosamine synthesis pathway in muscles of insulin resistant ob/ob mice. AB - Enhanced glucose flux via the hexosamine biosynthetic pathway has been implicated in insulin resistance. We measured products of this pathway, UDP-N-acetyl hexosamines (UDP-HexNAc), and activity of the rate-limiting enzyme L-glutamine:D fructose-6-phosphate amidotransferase (GFAT) in tissues of ob/ob mice and lean controls. Ob/ob mice were obese, hyperglycemic, and hyperinsulinemic. Resistance to the effect of insulin on glucose transport was demonstrated in isolated soleus muscles, although total GLUT-4 concentration was mildly increased in muscles from ob/ob mice. UDP-HexNAc concentrations in hindlimb muscles decreased between 8 and 17 wk but were always higher in ob/ob vs. controls (P < 0.001, mean increase 67%). Concentrations of UDP-hexoses and GDP-mannose were similar in ob/ob and control muscles. Muscle GFAT activity declined with age but was increased in ob/ob vs. controls at each age examined (P < 0.001, mean increase 108%). UDP HexNAc concentrations and GFAT activity were similar in livers of ob/ob and controls. These data suggest that glucose flux via the hexosamine pathway is selectively increased in muscle but not liver of ob/ob mice and may contribute to muscle insulin resistance in this model of non-insulin-dependent diabetes mellitus. PMID- 9227456 TI - Fate of insulin analogs in intact and nephrectomized rats determined by their receptor binding constants. AB - After intravenous injection of 125I-labeled human insulin and analogs in normal and nephrectomized rats, we examined their kinetic fate by Q-Sepharose separation into intact ligand, "fragments" (genuine fragments and protein-bound radioactivity), and iodide. Receptor binding association and dissociation constants (kass and kdis, respectively) of the analogs were estimated dynamically in vitro by BIAcore. The very fast disappearance of intact ligand from serum was found to be determined by 1) both kass and kdis of receptor-bearing tissue, thus substantiating our primary hypothesis; 2) elimination by kidneys, and 3) fast extravascularization. The rate of appearance of degradation products from receptor-mediated intracellular processing seems determined by kdis. With the possible exception of a truncated analog, ligand appears protected against degradation while the intracellular receptor-ligand complex remains intact. Non receptor-mediated processing in kidneys is slow, compared with the receptor mediated uptake and degradation of ligands with rate constants comparable to those of insulin. We observed binding of insulin and analogs putatively to serum proteins; binding capacity and affinity appeared insignificant for insulin but considerable for some analogs. PMID- 9227457 TI - Oxidative metabolism in insulin-treated gestational diabetes mellitus. AB - To investigate whether protein, carbohydrate, and fat metabolism was normalized in insulin-treated gestational diabetes mellitus (GDM), eight Hispanic women with GDM and eight healthy controls were studied at 32-36 wk of gestation and 6 wk postpartum. Net substrate utilization was measured using room respiration calorimetry. Exogenous substrate oxidation was determined by 13C recovered in breath CO2 from 13C-labeled leucine, glucose, and Hiolein. Women with GDM had higher 24-h oxygen consumption, carbon dioxide production, total energy expenditure, and basal metabolic rates than controls due to larger body mass. Adjusted for weight or fat-free mass, total energy expenditure, basal metabolic rate, and basal and 24-h whole body net protein, carbohydrate, and fat utilization did not differ between insulin-treated GDM subjects and controls in pregnancy or postpartum. Oxidation of [13C]leucine and [13C]glucose did not differ by group or pregnancy status. Recovery of exogenously administered [13C]Hiolein, a biosynthetic triglyceride, as breath 13CO2 was significantly lower in the GDM group antepartum and postpartum (P = 0.02), indicating lower oxidation of exogenous triglycerides in GDM. PMID- 9227458 TI - Abdominal adiposity rather than age and sex predicts mass and regularity of GH secretion in healthy adults. AB - We tested the hypothesis that body composition is the major predictor of growth hormone (GH) secretion in nonobese adults. We measured lean and fat tissue distribution (computerized tomography and dual-energy X-ray absorptiometry scan) and physical fitness [maximal oxygen consumption (Vo2max)] in 42 healthy nonobese adults (22 women and 20 men, age range 27-59 yr, mean +/- SE body mass index = 24 +/- 0.5 kg/m2). Deconvolution analysis was used to estimate specific features of 24-h GH secretion and clearance. Approximate entropy was used to quantify the regularity of GH release. Older subjects exhibited decreased estimates of GH secretion compared with younger subjects. Females had higher estimates of GH secretion, a longer GH half-life, and displayed more irregularity in GH release than males. Mean 24-h serum GH concentrations correlated inversely with intra abdominal fat and waist-to-hip ratio and positively with Vo2max. Multiple linear regression analysis revealed intra-abdominal fat as the dominant determinant of estimates of GH secretion. Vo2max was more important than sex and age in predicting GH secretion. We conclude that abdominal fat is the major determinant of GH secretion in healthy nonobese adults. Although the underlying mechanisms remain elusive, our findings extend the clinical implications of visceral adiposity to include hyposomatotropism. PMID- 9227459 TI - Extracellular ATP as an autocrine/paracrine regulator of prolactin release. AB - Recent evidence demonstrates that ATP, costored with a number of hormones and neurotransmitters in secretory granules, is coreleased during exocytosis of these agents. Here, we explored the possibility that extracellular ATP subserves an autocrine and/or paracrine role in the regulation of prolactin (PRL) release by subjecting rat pituitary cells to various experimental manipulations aimed at evaluating putative interactions between ATP and mammotropes. Our results strongly support the view that ATP functions as a local regulator of PRL secretion. To be more specific, we observed that ATP is released in a predictable manner by physiologically relevant secretagogues that are reasonably targeted to mammotropes. Moreover, we found that ATP can act directly on pituitary cells to stimulate the release of PRL from most (if not all) mammotropes. Finally, we determined that antagonism or removal of ATP leads to a diminution of PRL export from pituitary cells cultured under basal or thyrotropin-releasing hormone stimulated conditions. On the basis of these results, we propose that ATP acts locally to amplify and prolong the PRL secretory response elicited by a more traditional hypophysiotropic signal. PMID- 9227460 TI - Hepatic fatty acid synthase gene transcription is induced by a dietary copper deficiency. AB - A dietary copper (Cu) deficiency is associated with a twofold increase in hepatic fatty acid biosynthesis. We hypothesized that the induction of hepatic lipogenesis associated with a dietary Cu deficiency reflected an enhanced expression of genes encoding lipogenic enzymes, i.e., fatty acid synthase (FAS). Male weanling rats were pair-meal fed for 42 days a high-sucrose diet that was Cu deficient (CuD; 0.7 microgram Cu/g) or Cu adequate (CuA; 5.0 micrograms Cu/g). The CuD diet increased FAS enzymatic activity twofold (P < 0.05). This rise in enzymatic activity was accompanied by a threefold increase in FAS mRNA and a 2.5 fold increase in FAS gene transcription (P < 0.05). Neither the mRNA abundance nor the rate of gene transcription for phosphoenolpyruvate carboxykinase or beta actin was affected by the CuD diet. The induction of FAS gene transcription was associated with a 65-85% increase in hepatic reduced glutathione (GSH; P < 0.05). When hepatic GSH synthesis was suppressed by treating CuD rats with L-buthionine sulfoximine, the induction of FAS expression was completely prevented. Similarly, feeding N-acetylcysteine to CuA rats increased hepatic GSH levels 2.5-fold, and this was accompanied by a significant induction in FAS expression. These data indicate that the increase in hepatic lipogenesis associated with a Cu deficiency reflects an induction in hepatic lipogenic gene transcription (i.e., FAS) and that the rate of gene transcription may be dependent on hepatic thiol redox. PMID- 9227461 TI - Lipolytic responsiveness to epinephrine in nondiabetic and diabetic humans. AB - To determine whether the sensitivity of adipose tissue lipolysis to catecholamines is increased in poorly controlled insulin-dependent diabetes, the lipolytic response to epinephrine was measured in seven nondiabetic volunteers and seven poorly controlled diabetic subjects with use of [1-(14)C]palmitate as a tracer. Subjects received sequential 1-h infusions of epinephrine, which produced epinephrine concentrations of approximately 1,000, approximately 1,750, approximately 3,500, and approximately 6,000 pmol/l. A pancreatic clamp was used to maintain constant plasma hormone levels. Concentration-response curves were constructed for each subject from the integrated lipolytic response during each epinephrine infusion. There was no difference in maximal lipolytic response (117 +/- 19 vs. 152 +/- 11 mumol.kg-1.h-1) or in maximally effective (3,171 +/- 267 vs. 3,357 +/- 349 pmol/l) or half-maximally effective (1,081 +/- 109 vs. 1,015 +/ 120 pmol/l) epinephrine concentrations between nondiabetic and diabetic subjects, respectively (all P = NS). In control subjects, maximum beta hydroxybutyrate concentrations were achieved at lower epinephrine concentrations than those required for a maximum lipolytic effect. Thus, under pancreatic clamp conditions, the lipolytic response to epinephrine in nondiabetic and diabetic subjects was similar. PMID- 9227462 TI - An evaluation of the cross-linking model for the interaction of insulin with its receptor. AB - The cross-linking model for insulin receptor interactions, in which a single insulin molecule may form a cross-link between an insulin receptor's alpha subunits, has been expressed as a formal compartmental model and subjected to a systematic analysis, examining a number of predictions that have been made for this model. The kinetic parameters for the model were obtained by matching data from insulin receptor equilibrium binding studies and rates of formation of the insulin receptor complex. This analytical study has allowed a clear description of the kinetics of the ligand receptor complexes involved in such a mechanism. We conclude that the cross-linking model accounts for the anomaly of the 10-fold concentration difference in high- and low-affinity binding sites found when insulin binding is analyzed by conventional means. However, the phenomenon of acceleration of dissociation of labeled ligand by unlabeled ligand cannot be accounted for as an intrinsic part of the model. We suggest that this phenomenon arises from the destabilization of cross-link formation when a second insulin molecule binds. PMID- 9227463 TI - Molecular aspects of hepatobiliary transport. AB - Generation of bile flow is a regulated, ATP-dependent process and depends on the coordinated action of a number of transporter proteins in the sinusoidal and canalicular domains of the hepatocyte. Dysfunction of any of these proteins leads to retention of substrates, with conjugated hyperbilirubinemia or cholestasis as a result. In recent years many of the transport proteins involved in bile formation have been identified, cloned, and functionally characterized. The hepatocyte sinusoidal membrane contains transport proteins for the hepatic uptake of organic anions and cations and for the uptake of bile acids. The multispecific organic anion transporting polypeptide (OATP) mediates the hepatic uptake of organic anions and a variety of organic amphiphilic compounds, including organic cations. The organic cation transporter OCT1 more specifically transports small organic cations. NTCP is the Na(+)-bile acid cotransporting protein that mediates the hepatic uptake of bile acids. The canalicular transport proteins are able to transport endogenous and exogenous metabolites into the bile against steep concentration gradients. Most of these transporters are members of the large ATP binding cassette (ABC) superfamily, and their transport function directly depends on the hydrolysis of Mg2+/ATP. At least five ABC transporter proteins have been characterized so far: 1) the human multidrug resistance protein MDR1 mediates the excretion of hydrophobic, mostly cationic, metabolites; 2) MDR3 is involved in phosphatidylcholine secretion; 3) the canalicular bile acid transporter cBAT mediates secretion of monovalent bile salts and provides the molecular basis of bile acid-dependent bile flow; 4) SPGP, product of the P-glycoprotein sister gene, is exclusively expressed in the liver but its function is currently unknown; and 5) the human multidrug resistance protein MRP2 mediates the excretion of multivalent anionic conjugates. PMID- 9227464 TI - Ion pumps in epithelial cells: sorting, stabilization, and polarity. AB - The P-type family of ion-transporting ATPases includes all of the isoforms of Na(+)-K(+)-adenosinetriphosphatase (ATPase), the plasma membrane and organellar Ca(2+)-ATPases, gastric H(+)-K(+)-ATPase, and the recently characterized nongastric H(+)-K(+)-ATPases, among others. In epithelial cells, members of this pump family generate the cation gradients responsible for vectorial fluid and solute transport. To carry out this function, each P-type ATPase must be restricted to a specific membrane domain. Newly synthesized ion pumps must be sorted to their appropriate destinations and retained there after their delivery. Recently, progress has been made toward understanding the signals and targeting pathways that epithelial cells employ to generate anisotropic pump distributions. The mechanisms involved in generating these pump distributions may serve as well to regulate pump function. PMID- 9227465 TI - Sodium-dependent nucleoside transport in the human intestinal brush-border membrane. AB - The objective of the study was to determine the identity and kinetic characteristics of nucleoside transporters present in the brush-border membrane of the human jejunum. With use of brush-border membrane vesicles, uptake of [3H]uridine was stimulated two- to threefold by an inwardly directed Na+ gradient and was inhibited by both 100 microM thymidine and 100 microM guanosine nucleosides, which serve as model substrates for purine (N1, cif) and pyrimidine (N2, cit) transporters, respectively. [3H]thymidine and [3H]guanosine transport exhibited an overshoot phenomenon only in the presence of a Na+ gradient. Na(+) thymidine uptake was inhibited by 100 microM cytidine or thymidine but not by guanosine, inosine, formycin B, or hypoxanthine. [3H]guanosine uptake was inhibited by 100 microM inosine, guanosine, or formycin B but not by thymidine or cytidine. Both adenosine and uridine inhibited uptake of [3H]thymidine and [3H]guanosine to a similar extent, indicating that both N1, cif and N2, cit Na(+) nucleoside transporters are expressed in human jejunum. Enhanced uptake of Na(+) thymidine by an inside-negative potential difference generated by K+ and valinomycin provides evidence that nucleoside transport is rheogenic, involving net transfer of a positive charge. The Hill coefficient was unity for all three substrates, indicating a Na(+)-nucleoside coupling stoichiometry of 1:1. At saturating Na+ concentration (150 mM) the kinetic parameters (n = 3-4) Michaelis Menten constant and maximum velocity for uridine, thymidine, and guanosine uptake were 4.15 +/- 1.79, 2.74 +/- 0.58, 12.02 +/- 1.34 microM and 25.93 +/- 7.38, 16.10 +/- 3.64, 63.92 +/- 10.23 pmol.mg-1.10 s-1, respectively. These results suggest that, in contrast to the human kidney that expresses the N4 nucleoside transporter, the human jejunum expresses both N1 and N2 Na(+)-nucleoside transporters. PMID- 9227466 TI - Regional differences in cholinergic activity of muscle fibers from the human gastroesophageal junction. AB - Muscles of the gastroesophageal junction that contribute to the lower esophageal sphincter (LES) include clasplike semicircular fibers on the right and slinglike oblique gastric fibers on the left. This study examined whether in vitro differences between the sling and clasp muscles could account for the in vivo asymmetry of LES pressure and its cholinergic contribution. Isometric tension was recorded from muscle strips of the sling, clasp, and circular layers of the esophagus and gastric fundus isolated from surgical specimens. The sling developed less spontaneous tension (8.9 +/- 4.3 mN/mm2) than the clasp (25.0 +/- 7.4 mN/mm2, P < 0.01) but showed a fivefold greater increase in response to carbachol. Eserine (1 microM) increased tension in the sling muscle (64.5 +/- 29.7%), but not in the clasp, whereas 1 microM atropine or 1 microM tetrodotoxin had no significant effect in either muscle. In both muscles, tension was reduced by 10 microM sodium nitroprusside. Sling or clasp muscle differed from circular muscle of the esophagus or gastric fundus in spontaneous tension, carbachol response, or responses to electrical stimulation. Thus the clasp muscle develops greater spontaneous tension, whereas the sling is more sensitive to cholinergic stimulation, providing a potential explanation for the in vivo asymmetry of the LES pressure and its response to cholinergic blockade. PMID- 9227467 TI - Isolation and culture of bovine pancreatic duct epithelial cells. AB - We describe a method to isolate and culture epithelial cells from the main duct of the bovine pancreas. In primary cultures, secretin caused a dose-dependent increase in intracellular adenosine 3',5'-cyclic monophosphate (cAMP) and stimulated electrogenic transepithelial ion transport. Elevation of intracellular cAMP increased the rate coefficient for 36Cl- efflux from 0.14 +/- 0.03 to 0.47 +/- 0.12 min-1, and plasma membrane conductance, measured by the whole cell patchclamp technique, was increased from 0.7 +/- 0.1 to 6.9 +/- 0.8 nS. The cAMP activated anion currents had properties similar to those mediated by the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Cells grown on permeable supports formed confluent monolayers with high transepithelial electrical resistance (1.004 +/- 96 omega. cm2) and generated a lumen negative transepithelial voltage difference (-2.5 +/- 0.6 mV). The short-circuit current (Isc) was increased by forskolin or secretin and was inhibited 87 +/- 4% by addition of ouabain (100 microM) to the basolateral bathing solution. Replacement of bathing solution Cl- by cyclamate reduced the forskolin-induced steady-state increase in Isc from 5.3 +/- 0.5 to 0.2 +/- 0.2 microA/cm2, suggesting that the stimulated current is due to anion secretion. The results of these studies demonstrate that large numbers of pancreatic ductal cells can be isolated and grown in primary cell culture. The monolayers express differentiated functions and will be useful for studies of acute and chronic regulation of ion transport in pancreatic duct epithelial cells. PMID- 9227468 TI - IL-1 beta induces synthesis of phospholipase A2-activating protein in rabbit distal colon. AB - In inflammatory bowel disease, the colonic mucosa is infiltrated by inflammatory cells that secrete a variety of inflammatory mediators such as interleukin-1 beta (IL-1 beta). IL-1 beta caused a delayed increase in Cl- secretion and in prostaglandin E2 (PGE2) release in rabbit distal colon. Both of these effects were abolished with cycloheximide, implying a role for protein synthesis in mediating IL-1 beta's effect. With the use of Western blot assays, the protein was identified as the phospholipase A2 (PLA2)-activating protein (PLAP). IL-1 beta caused a concentration-dependent and a time-dependent increase in PLAP levels as well as in PLA2 activity, with the maximal increase observed at an IL-1 beta concentration between 10 and 30 ng/ml reached in 2-10 min. The PLAP mRNA levels were also regulated by IL-1 beta with a similar time course. PLAP is constitutively present in the epithelial cells and in the subepithelial layer of the distal colon. These findings suggest a direct effect of IL-1 beta on intestinal epithelial cells to cause an increase in PLAP levels and phospholipase A2 activity and subsequent increase in PGE2 levels. PMID- 9227469 TI - Salutary effects of ATP-MgCl2 on altered hepatocyte signal transduction after hemorrhagic shock. AB - Although previous studies indicate that hepatocyte P2 purinoceptors, macrophage adenosine 3',5'-cyclic monophosphate (cAMP), and beta-adrenergic receptors decrease after hemorrhage and that administration of ATP-MgCl2 after hemorrhage normalizes these parameters, it is not known whether other aspects of hepatocyte signal transduction processes, such as transmembrane coupling, are also affected by hemorrhage and, if so, whether ATP-MgCl2 has any beneficial effects on signal transduction. To study this, rats underwent a 5-cm midline laparotomy (i.e., trauma induced) and were bled to and maintained at a mean arterial pressure of 40 mmHg until 40% of a maximum bleed-out volume was returned in the form of Ringer lactate (RL). They were then resuscitated with three times the volume of shed blood with RL over 45 min followed by two times the volume with RL+ATP-MgCl2 (50 mumol/kg body wt) or an equivalent volume of saline over 95 min. Hepatocytes were isolated at 4 and 27 h after resuscitation, and basal as well as stimulated levels of cAMP and inositol 1,4,5-trisphosphate (IP3) were determined. The results indicate that basal levels of cAMP decreased whereas IP3 increased after hemorrhage and resuscitation. Receptor-dependent stimuli (i.e., glucagon and vasopressin) failed to elicit cAMP or IP3 accumulation after hemorrhage. In contrast, receptor-independent stimulation was not impaired. ATP-MgCl2 treatment, however, prevented the decreased basal levels of cAMP and IP3 and the ability of hepatocytes to respond to receptor-dependent stimulation. Thus ATP-MgCl2 treatment of animals after trauma-hemorrhage and resuscitation attenuates the impaired second messengers cAMP and IP3 and their membrane transduction processes. PMID- 9227470 TI - Role of iron in NF-kappa B activation and cytokine gene expression by rat hepatic macrophages. AB - A redox-sensitive nuclear factor, NF-kappa B, induces transcription of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in macrophages. The present study has investigated the role of iron in NF-kappa B activation and TNF alpha and IL-6 expression by rat hepatic macrophages (HM). As an in vivo model, cholestatic liver injury was induced in rats by ligation of the common bile duct (BDL). During the first 2 wk after BDL, there was an increase in the hepatic level of thiobarbituric acid-reactive substances (TBARS) that was accompanied by the appearance of protein-malondialdehyde adducts in the periportal region. This increase was reduced after 3 wk. TNF-alpha and IL-6 mRNA levels in HM from the BDL rats were increased at 1 and 2 wk and attenuated at 3 wk. Gel mobility shift assay of HM nuclear extracts demonstrated the similar temporal pattern of enhanced NF-kappa B binding activity. Treatment of the BDL animals with 1,2 dimethyl-3-hydroxypyrid-4-one (L-1), a lipophilic iron chelator, suppressed the increases in hepatic TBARS by 64%, plasma alanine aminotransferase by 45%, and HM TNF-alpha and IL-6 mRNA by > 84%. Concomitantly, the HM NF-kappa B binding activity was reduced close to the level observed in sham-operated rats. Treatment of cultured HM with L-1 also blocked lipopolysaccharide-stimulated NF-kappa B activation and TNF-alpha and IL-6 expression at mRNA and protein levels. These results demonstrate that the iron chelator effectively blocks NF-kappa B activation and coordinate TNF-alpha and IL-6 gene upregulation by HM in cholestatic liver injury or under in vitro lipopolysaccharide stimulation. These findings support a pivotal role for iron in activation of NF-kappa B and cytokine gene expression by HM in vitro and in vivo. PMID- 9227472 TI - Intracisternal antisense oligonucleotides to TRH receptor abolish TRH-evoked gastric motor excitation. AB - Thyrotropin-releasing hormone (TRH) from the nucleus raphe obscurus (nROb) innervates the dorsal vagal complex (DVC) and activates gastric motor function. Assessment of the importance of TRH has been hampered by the lack of TRH receptor antagonists. To overcome this, rats were given intracisternal antisense oligonucleotides against the first 18 bases of TRH receptor mRNA, mismatch oligonucleotides, or saline. Rats were anesthetized, and L-glutamate (15 nmol), TRH (1 and 10 pmol), and saline were microinjected into the DVC and nROb while gastric motor function was monitored. Intracisternal TRH mRNA antisense oligonucleotides abolished the gastric excitatory affects of microinjection of TRH, but not L-glutamate, into the DVC, and the response to TRH recovered after 2 wk of no antisense treatment. Chemical stimulation of the nROb increased intragastric pressure in saline- and mismatch- but not antisense-treated animals. These studies demonstrate that intracisternal TRH receptor antisense oligonucleotides produce a selective and reversible "knockdown" of responsiveness to exogenous TRH in the DVC, as well as to excitation of an endogenous TRH pathway controlling gastric function. It also provides a new tool for assessment of TRH pathways in hindbrain control of gastric function. PMID- 9227471 TI - Cholecystokinin depolarizes guinea pig sphincter of Oddi neurons by activating CCK-A receptors. AB - Motility studies indicate that cholecystokinin (CCK) acts through a neural mechanism in the sphincter of Oddi (SO) after meals. To evaluate its actions in SO ganglia, CCK was applied by microejection (0.1 mM) or superfusion (0.1 to 300 nM) while recording was carried out intracellularly from intact SO neurons. In tonic cells, microejection and superfusion of CCK caused a prolonged depolarization accompanied by action potentials. In phasic cells, microejection of CCK caused brief and/or prolonged depolarizations, but superfusion caused only prolonged depolarizations. In afterhyperpolarized cells, CCK did not cause a detectable change in the resting membrane potential. In low-Na+ Krebs solution, the prolonged depolarizations in both tonic and phasic cells were significantly reduced. Unsulfated CCK (100 nM) had no effect. CCK-induced depolarization was significantly reduced by a CCK-A, but not a CCK-B, receptor antagonist. It is concluded that CCK can act on CCK-A receptors to depolarize SO neurons. However, it is unlikely that hormonal CCK could mediate such an action because of the discrepancy between the sensitivity of SO neurons for CCK and the peak concentrations of CCK in the serum after a meal. PMID- 9227473 TI - Synthesis of citrulline and arginine from proline in enterocytes of postnatal pigs. AB - The synthesis of ornithine, citrulline, and arginine from proline was quantified in enterocytes of newborn (0-day-old) pigs, 2- to 21-day-old suckling pigs, and 29- to 58-day-old pigs weaned at 21 days of age. Mitochondria were prepared from enterocytes for measurement of proline oxidase activity. For metabolic studies, cells were incubated at 37 degrees C for 30 min in Krebs bicarbonate buffer (pH 7.4) containing 5 mM D-glucose and 0.0-5.0 mM L-[U-14C]proline with or without 2.0 mM L-glutamine. Proline oxidase activity and rates of synthesis of citrulline and arginine from proline were high in enterocytes of newborn pigs and markedly decreased in 7-day-old pigs. With increasing piglet age from 7 to 21 days, enterocyte proline oxidase activity and rate of conversion of proline into citrulline progressively increased, but the values in 21-day-old pigs remained much lower than in newborn pigs. The synthesis of ornithine, citrulline, and arginine from 0.5-5 mM proline was concentration dependent and required the addition of glutamine. About 80-90% of utilized proline carbons were recovered in ornithine plus citrulline plus arginine, with CO2 being a minor product. These results demonstrate a hitherto unrecognized pathway for the synthesis of ornithine, citrulline, and arginine via proline oxidase in enterocytes and challenge the current concept that pyrroline-5-carboxylate (P5C) is synthesized in the small intestine only from glutamine/glutamate via P5C synthase on the basis of previous work with postnatal rats. PMID- 9227475 TI - Characterization of intracellular copper pools in rat hepatocytes using the chelator diamsar. AB - When hepatocytes are incubated with the chelator diamsar, two pools can be identified, which we have termed extractable and nonextractable. On entering the hepatocyte, 67Cu first associates with the extractable pool and, after approximately 2 h, moves to the nonextractable pool. Both pools demonstrate saturation and are filled as a function of Cu concentration and incubation time. Using the Michaelis-Menten equation, we have estimated the size of the pools after incubation with 67Cu for 30 min and 4 h. During this period the extractable pool decreases in size from 200 +/- 27 to 116 +/- 5 pmol/microgram DNA, whereas the nonextractable pool increases from 28 +/- 9 to 77 +/- 11 pmol/microgram DNA. Movement of Cu from the nonextractable pool to the extractable pool is slow and incomplete. Using [3H]diamsar, we demonstrate that uptake of the chelator is not rate limiting and probably does not occur by pinocytosis. Incubation with diamsar does not affect the activity of superoxide dismutase or cytochrome-c oxidase, although it does prevent the incorporation of 67Cu into ceruloplasmin. Incubation with zinc, which induces metallothionein, results in an increase in 67Cu associated with the nonextractable pool, suggesting that 67Cu-metallothionein constitutes at least part of the nonextractable pool. PMID- 9227474 TI - Characterization of PACAP receptors and signaling pathways in rabbit gastric muscle cells. AB - Pituitary adenylate cyclase-activating peptide (PACAP) receptors and their signaling pathways were characterized in dispersed rabbit gastric muscle cells. 125I-PACAP-27 and 125I-vasoactive intestinal peptide (VIP) binding to muscle cells were inhibited equally by PACAP and VIP (mean inhibitory concentration 0.8 to 1.3 nM) and desensitized to the same extent (70-80%) by exposure to either peptide. PACAP, like VIP, increased cytosolic free Ca2+ and the formation of L [3H]citrulline, NO-3/NO-2, guanosine 3',5'-cyclic monophosphate (cGMP), and adenosine 3'5'-cyclic monophosphate (cAMP) and induced relaxation (mean effective concentration 1.8 +/- 0.1 nM) that was partly inhibited by NG-nitro-L-arginine (L NNA), VIP-(10-28), and PACAP 6-38. L-[3H]citrulline and cGMP formation were blocked by nifedipine, L-NNA, and pertussis toxin (PTx), implying activation of a G protein-coupled, Ca(2+)-calmodulin-dependent nitric oxide (NO) synthase. PACAP induced relaxation was inhibited to the same extent (46-49%) by nifedipine, L NNA, PTx, and the protein kinase G inhibitor KT-5823; the inhibition reflected the component of relaxation mediated by the NO-cGMP pathway. The residual relaxation was abolished by the protein kinase A inhibitor H-89. The pattern of inhibition of all responses was identical to that observed with VIP. Desensitization with VIP or PACAP abolished cAMP formation but had no effect on L [3H]citrulline and cGMP formation induced by either peptide. Receptor protection with VIP or PACAP preserved fully all responses (L-[3H]citrulline, cGMP, and cAMP formation and relaxation) to either peptide. The complete cross-competition, cross-desensitization, cross-antagonism, and cross-protection of receptors by either VIP or PACAP are consistent with interaction of both peptides with the same receptors; the receptors consist of two classes, each coupled to a distinct signaling pathway. PMID- 9227476 TI - Evidence for the existence of a carrier-mediated folate uptake mechanism in human colonic luminal membranes. AB - Colonic bacteria synthesize significant amounts of folate. The current studies were undertaken to examine the presence of a possible folate transporter and its characterization in apical membranes of the human colon. Apical membrane vesicles were purified from mucosal scrapings of proximal organ donor colons using a differential centrifugation and divalent cation precipitation technique. [3H]folate (PteGlu) uptake was measured by a rapid filtration technique. Our results demonstrate that [3H]PteGlu uptake into these vesicles 1) was significantly increased with decreasing pH of the incubation buffer, 2) was markedly inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS) and 4-acetamido-4'-isothiocyanostilbene-2,2'-disulfonic acid but not by acetazolamide, amiloride, bumetanide, furosemide, and diphenyl 2-carboxylic acid, 3) was sensitive to temperature and osmolarity of the incubation medium, 4) was inhibited by structural analogs methotrexate and 5-methyltetrahydrofolate, 5) exhibited trans-stimulation phenomenon, 6) was potential insensitive, and 7) exhibited saturation kinetics with an apparent Michaelis constant for PteGlu of 8.2 +/- 2.4 microM and a maximal enzyme reaction rate of 19.8 +/- 2.9 pmol.mg protein-1.10 s-1. Studies on distal colonic membranes also demonstrated the presence of a folate transporter with similar kinetic characteristics. These results demonstrate the existence of a pH-dependent, DIDS-sensitive, electroneutral carrier-mediated mechanism for folate absorption in the human colon. PMID- 9227477 TI - Carrier-mediated transport of conjugated bile acids across the basolateral membrane of biliary epithelial cells. AB - When secreted into bile, unconjugated dihydroxy bile acids are absorbed passively by cholangiocytes according to the cholehepatic circulation hypothesis. A fraction of these are likely to be conjugated during transcellular transport. Experiments were performed using fluorescent conjugated bile acids to test whether carrier-mediated transport of conjugated bile acids is present in the basolateral domains of polarized cholangiocytes of intrahepatic bile ductules isolated from rat liver. The time course of the cellular localization of cholyl NBDAB-Gly and chenodeoxycholyl-NBDAB-Gly, which are anionic fluorescent derivatives of the corresponding glycine-conjugated bile acids, was characterized using an image-analysis system. With 0.3-3 microM solutions, fluorescence was present at 1 and 3 min in the basolateral area of cholangiocytes. Staining in the apical region occurred later, with a peak after 15 min of incubation. The basolateral uptake of the two fluorescent bile acids was temperature dependent and Na+ independent, and was not influenced by the addition of amiloride, by lowering of the medium pH to 6.0, or by preincubation with valinomycin. Uptake was partially inhibited by the absence of Cl- or HCO3- in the perfusate, by preincubation with 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), and by the presence of different organic anions or unconjugated and conjugated bile acids in the medium. When cells were preloaded with an ethyl ester of chenodeoxycholyl-NBDAB-Gly, which is hydrolyzed by intracellular esterases, the decrease of cell fluorescence was partly inhibited by H2DIDS, whereas it was stimulated by the presence of 20 microM cholyltaurine in the medium. It is concluded that transport of conjugated bile acid anions across the basolateral membrane of the polarized rat cholangiocyte is carrier mediated. The conjugated bile acid transporter is likely to be an anion exchanger and is likely to be involved in bile secretion whenever conjugated bile acids or other organic anions are transported from the base of the biliary ductular epithelial cells into the plasma of the periductular capillary plexus. PMID- 9227478 TI - Insulin induces Ca2+ influx into isolated rat hepatocyte couplets. AB - Isolated rat hepatocyte couplets were used to study the direct effect of insulin on intracellular Ca2+ homeostasis. Insulin induced a dose-dependent increase in hepatocellular Ca2+ that was gradual, generally monophasic, and reversible. Chelation of extracellular Ca2+ abolished the insulin-induced Ca2+ response, and this suppression was not related to an effect on insulin binding, as indicated by displacement studies. We thus tested the effect of several Ca2+ channel inhibitors on insulin-induced Ca2+ influx. Verapamil at 20 or 200 microM was without effect, whereas 500 microM nickel and 50 microM gadolinium strongly inhibited insulin-induced Ca2+ entry. Finally, we tested whether insulin-induced Ca2+ movements were implicated in the stimulation of mitogen-activated protein kinase (MAPK) activity, which we measured with the use of an immune-complex assay. Verapamil was without effect on the insulin-dependent stimulation of p44mapk activity, whereas addition of ethylene glycol-bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid, nickel, or gadolinium strongly inhibited the effect of the peptide hormone. Our results indicate that insulin triggers Ca2+ influx into hepatocytes, possibly through the opening of channels on the plasma membrane, and that this effect is important for insulin activation of MAPK. PMID- 9227479 TI - Dual effects of PACAP on guinea pig gallbladder muscle via PACAP-preferring and VIP/PACAP-preferring receptors. AB - The aims of this study were to determine the effect and mechanism of action of pituitary adenylate cyclase-activating peptide (PACAP) on gallbladder muscle. Guinea pig gallbladder muscle strips were studied isometrically. In noncontracted muscle strips, PACAP-27 and PACAP-38 caused dose-dependent contractions, whereas vasoactive intestinal peptide (VIP) caused dose-dependent relaxation. PACAP-27 contractions were resistant to tetrodotoxin, atropine, and the substance P receptor antagonist [D-Arg1,D-Trp7,9,Leu11]substance P (Spantide) but were inhibited by the selective PACAP receptor antagonist PACAP-(6-38) and slightly increased with the VIP receptor antagonist [4-chloro-D-Phe6,Leu17]VIP. In cholecystokinin-precontracted muscle strips, both VIP and PACAP caused relaxations. This relaxant effect of PACAP-27 was inhibited by PACAP-(6-38) and [4-chloro-D-Phe6,Leu17]VIP, but not by tetrodotoxin. These studies suggest that PACAP has dual excitatory and inhibitory effects on guinea pig gallbladder muscle. The contractile effect of PACAP is a direct action on muscle through PACAP-preferring receptors. The relaxant effect of PACAP is seen in precontracted muscle strips and mediated through VIP/ PACAP-preferring receptors. PMID- 9227480 TI - Reactive oxygen species and calcium homeostasis in cultured human intestinal smooth muscle cells. AB - Reactive oxygen species (ROS) significantly alter cell function. We examined the effects of hydrogen peroxide (H2O2) and xanthine/xanthine oxidase (X/XO) on isolated intestinal muscle cells. We assessed cell viability with the exclusion dye trypan blue and assayed the effects of H2O2 and X/XO on the intracellular redox state with the fluorescent probe 2',7'-dichlorofluorescein. Intracellular calcium concentration was measured in cells loaded with fura 2-acetoxymethyl ester, and we recorded whole membrane currents with conventional patch-clamp methods. Cells remained viable after a 5-min exposure to H2O2 and X/XO. H2O2 and X/XO led to a significant rise of the intracellular concentration of ROS. H2O2 (270 microM to 2.7 mM) as well as X/XO (0.25-16 mU; 0.5 mM xanthine) significantly increased intracellular calcium concentrations. Depletion of intracellular calcium with ryanodine or thapsigargin did not abolish the effect of ROS on the intracellular calcium concentration. In the absence of external calcium or in the presence of the calcium channel blocker nifedipine, H2O2 and X/XO still increased the intracellular calcium level. Thus calcium influx and calcium release from internal stores contributed to this rise in cytosolic calcium. Catalase and superoxide dismutase blunted or completely abolished the changes in calcium concentration elicited by H2O2 and X/XO. Exposure to ROS resulted in a rapid decline of the membrane resistance without significant changes in voltage-sensitive ion currents. We conclude that ROS disrupt the calcium homeostasis of cells at concentrations that do not lead to immediate cell death. The resulting elevation in cytosolic free calcium will activate a variety of biochemical reactions and may thus contribute to the cytotoxicity of reactive oxygen molecules. PMID- 9227481 TI - Effects of the inflammatory mediator prostaglandin E2 on myenteric neurons in guinea pig ileum. AB - The effects of the inflammatory mediator prostaglandin E2 (PGE2) on myenteric neurons were investigated by intracellular recordings in a conventional plexus preparation. Bath application of PGE2 (1-1,000 nM) evoked a concentration dependent and reversible slow depolarization and an augmentation of excitability in 23 of 26 AH and 12 of 13 S neurons. The amplitude of the slow depolarization ranged from 4 +/- 1 mV at 1 nM to 13 +/- 3 mV at 1 microM in S and AH neurons. In AH neurons, PGE2 evoked an increase in membrane resistance and a reduction of afterhyperpolarization. In S neurons, PGE2 evoked either an increase or a decrease in membrane resistance. PGE2 slightly reduced the amplitude of electrically evoked fast excitatory postsynaptic potentials and had no effect on slow excitatory postsynaptic potentials. Moreover, PGE2 evoked bursts of fast excitatory postsynaptic potentials and action potentials in S neurons, indicative of cyclical neural activity in the myenteric plexus. It is concluded that the inflammatory mediator PGE2 can act as an excitatory neuromodulator of gastrointestinal motility through direct action on neurons in the myenteric plexus. PMID- 9227482 TI - Adaptive Kupffer cell alterations after femur fracture trauma in rats. AB - Because Kupffer cells constitute the largest fixed macrophage population and reside at a strategic position in hepatic sinusoids, interacting with hepatocytes, circulating cells, and mediators from the gut, they may be important in the inflammatory response after injury. This study examined the effect of remote tissue injury on Kupffer cell function. Femurs of Sprague-Dawley rats were fractured under anesthesia. Subsequently, their livers were perfused for measurement of oxygen consumption and the isolation and culture of Kupffer cells. At 2 and 48 h after femur fracture, hepatic oxygen consumption increased 17 and 19%, respectively. Gadolinium chloride pretreatment to ablate Kupffer cells blocked this increase of hepatic oxygen consumption after femur fracture but had no effect in sham-operated animals. In Kupffer cells isolated and cultured 2 h after femur fracture, superoxide formation stimulated by phorbol ester increased eightfold, phagocytosis increased fourfold, and lipopolysaccharide (LPS) stimulated prostaglandin E2 increased sixfold in comparison to sham-operated controls. In contrast, LPS-stimulated tumor necrosis factor-alpha and nitric oxide production decreased 50 and 60%, respectively. These data show that peripheral trauma rapidly induces changes in hepatic macrophages characterized by adaptation to a more antimicrobial and less proinflammatory phenotype. PMID- 9227483 TI - Molecular and functional characterization of intestinal Na(+)-dependent neutral amino acid transporter B0. AB - We have cloned the Na(+)-dependent neutral amino acid transporter B0 (ATB0) from rabbit jejunum and from the human intestinal cell line Caco-2. Rabbit intestinal ATB0 (riATB0) cDNA codes for a protein of 541 amino acids with 10 potential transmembrane domains. When expressed in HeLa cells, riATB0 mediates the transport of several neutral amino acids, including glutamine, in a Na(+) dependent manner. Anionic amino acids, cationic amino acids, and N-methylated amino acids are excluded by riATB0. When expressed in Xenopus laevis oocytes, riATB0 increases the transport of neutral amino acids severalfold. The induced transport activity is specific for neutral amino acids, with no noticeable interaction with anionic, cationic, and N-methylated amino acids. However, riATB0 does interact with anionic amino acids at acidic pH. In oocytes expressing riATB0, the neutral amino acid threonine evokes inward currents at a holding potential of -50 mV. The amino acid-evoked current is sensitive to membrane potential. The inward current increases as the membrane potential is hyperpolarized, but the current reverses at about -30 to -40 mV. Threonine evokes outward currents if the membrane potential is depolarized beyond this value. We have also cloned the ATB0 from the human intestinal cell line Caco-2. The Caco-2 ATB0 cDNA also codes for a protein of 541 amino acids that is essentially identical to the ATB0 expressed in the human choriocarcinoma cell line JAR. Reverse transcription-polymerase chain reaction (RT-PCR) and restriction analysis of the RT-PCR products indicate that the human intestine and the human kidney proximal tubular cell line HKPT express an ATB0 identical to the ATB0 expressed in Caco-2 cells. PMID- 9227484 TI - Human spasmolytic polypeptide decreases proton permeation through gastric mucus in vivo and in vitro. AB - Exogenously administered trefoil peptides are gastroprotective in rat injury models. We hypothesized that trefoil-associated gastroprotection occurred by decreasing the rate of proton permeation through mucus. Gastric surface cell intracellular pH and mucus gel thickness were measured by in vivo microscopy. Gastric mucosal blood flow was measured by laser-Doppler flowmetry. The effect of human spasmolytic peptide (hSP) on H+ diffusion through 5% purified porcine mucin was measured using an Ussing chamber. Buffering action of mucin was measured by titration. In vivo, gastric mucosal blood flow and mucus gel thickness were not affected by any of the treatments. Topical hSP, but not intravenous hSP, decreased initial acidification rate and elevated the intracellular pH of gastric surface cells during luminal acid challenge. In in vitro studies, hSP dose dependently decreased the diffusion coefficient of H+ through 5% porcine mucin solution. hSP had no significant effect on the buffering action of mucin solutions. These data support our hypothesis that hSP interacts with gastric mucin in a manner that inhibits proton permeation through the mucus gel layer. PMID- 9227486 TI - Effect of oxidative stress on cellular functions and cytosolic free calcium of rat pancreatic acinar cells. AB - The present study evaluates the effect of free radicals generated by xanthine oxidase-catalyzed oxidation of hypoxanthine on cellular function of isolated rat pancreatic acinar cells. The results show that a rapid and sustained increase in intracellular Ca2+ concentration ([Ca2+]i) preceded all other morphological and functional alterations investigated. Radical-induced [Ca2+]i increase was largely inhibited by 3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester, which prevents Ca2+ release from intracellular stores, but not by Ca2(+)-depleted medium. Radicals released Ca2+ from thapsigargin-insensitive, ryanodine-sensitive intracellular stores, whereas the secretagogue caerulein at physiological concentrations mainly released Ca2+ from thapsigargin-sensitive stores. In contrast to effects of the secretagogue, radical-induced Ca2+ changes did not cause luminal protein secretion but cell death. In single-cell measurements, both secretagogue and radicals induced oscillations of [Ca2+]i. Radical-induced oscillations had a lower frequency but similar amplitude when compared with caerulein-induced oscillations. 1,2-Bis(2-aminophenoxy)ethane-N,N,N',N' tetraacetic acid and ryanodine, which prevented the radical-induced Ca2+ increase without altering the generation of radicals, markedly reduced the radical-induced cell damage. These results suggest that the Ca2+ increase mediates the radical induced cell injury. The studies also indicate that not only the extent and duration but also the origin of [Ca2+]i release as well as the frequency of Ca2+ oscillations may determine whether a pancreatic acinar cell will secrete or die. PMID- 9227485 TI - Somatostatin inhibits gastrin release and acid secretion by activating sst2 in dogs. AB - Somatostatin is a potent inhibitor of gastrin-stimulated acid secretion by activation of somatostatin receptor type 2 (sst2) in vivo, probably in part by blocking gastrin-stimulated histamine release from enterochromaffin-like cells expressing sst2. We propose that activation of sst2 may also regulate meal stimulated acid secretion by blocking gastrin release from antral G cells. Using peptide analogs relatively selective for sst2 (NC-8-12), sst3 (BIM-23058), and sst5 (BIM-23052), we tested this hypothesis in two ways: first, in vivo by measuring plasma gastrin release during meal-stimulated acid secretion in dogs, and second, in vitro by measuring bombesin-stimulated gastrin release from an enriched culture of canine antral G cells. In vivo, a low dose (0.05 nmol.kg-1.h 1) of NC-8-12 inhibited acid secretion 56 +/- 16% without blocking gastrin release. A higher dose (1 nmol.kg-1.h-1) of NC-8-12 abolished acid secretion and inhibited gastrin release by 61 +/- 4%, whereas the highest dose (5 nmol.kg-1.h 1) inhibited gastrin release by 84 +/- 3%. Only the highest doses (5 nmol.kg-1.h 1) of BIM-23058 and BIM-23052 significantly inhibited gastrin release and acid secretion. In vitro, NC-8-12 (10(-9) M) reduced bombesin-stimulated gastrin release from antral G cells by 49 +/- 5%, whereas BIM-23058 and BIM-23052 were at least 100-fold less effective. These results indicate that somatostatin activation of sst2, but not sst3 or sst5, is the major pathway for somatostatin induced inhibition of meal-stimulated gastrin release and acid secretion. PMID- 9227487 TI - Transcriptional regulation of human inducible nitric oxide synthase gene in an intestinal epithelial cell line. AB - The inducible form of nitric oxide synthase (iNOS) is expressed during inflammation of the intestine and may contribute to tissue injury. We have examined iNOS transcriptional regulation in DLD-1 cells, a human intestinal epithelial line that produces large amounts of nitric oxide and iNOS mRNA in response to a combination of the proinflammatory cytokines interleukin-1 beta (IL 1 beta) and interferon-gamma (IFN-gamma). Levels of iNOS mRNA are extremely low in unstimulated DLD-1 cells but increase dramatically after cytokine treatment. Nuclear run-on analyses demonstrated that transcriptional activation, which accounts for a portion of this increase, is dependent on both IL-1 beta and IFN gamma and requires de novo protein synthesis. Transfection of DLD-1 cells with reporter constructs containing deletions of the iNOS promoter showed that sequences located between 8.7 and 10.7 kb upstream of the transcription initiation site are necessary for cytokine responsiveness. This region contains potential binding sites for several cytokine-induced transcription factors and was shown to function in either orientation when placed upstream of a basal iNOS promoter segment terminating at-1.1 kb. The extremely distal location of the cytokine-responsive region contrasts with the reported positions of elements involved in the regulation of iNOS transcription in other cell types. Our data also suggest that posttranscriptional events could play a significant role in regulating iNOS gene expression in human intestinal epithelia. PMID- 9227488 TI - Asymmetry of lower esophageal sphincter pressure: is it related to the muscle thickness or its shape? AB - Lower esophageal sphincter (LES) pressure shows axial and circumferential asymmetry, the reasons for which are not clear. Our aim was to determine whether the muscle thickness and shape of the LES were the reasons for the axial and circumferential asymmetry in the LES pressure. High-frequency, catheter-based intraluminal ultrasonography was performed to obtain images of the human LES and esophagus. Station pull-through manometry was performed to record the axial and circumferential asymmetry of LES pressure. Circular and longitudinal muscle were thicker in the LES compared with in the esophagus. There was a close correlation between the axial asymmetry in LES pressure and circular muscle thickness. The thickness of LES muscle increased and decreased with an increase and decrease in the LES pressure, respectively. The circumferential asymmetry in resting LES pressure was related to the noncircular shape of the LES. A swallow-induced LES relaxation was followed by its contraction. During the contraction phase, the LES and esophagus showed relative symmetry in pressure and shape. We conclude that the axial asymmetry of LES pressure is explained by its muscle thickness. On the other hand, circumferential asymmetry is related to the noncircular shape of the LES. PMID- 9227489 TI - Augmentation of deglutitive upper esophageal sphincter opening in the elderly by exercise. AB - Earlier studies have shown that the cross-sectional area of the deglutitive upper esophageal sphincter (UES) opening in healthy asymptomatic elderly individuals is reduced compared with healthy young volunteers. The aim of this study was to determine the effect of a head-raising exercise on swallow-induced UES opening and hypopharyngeal intrabolus pressure in the elderly. We studied a total of 31 asymptomatic healthy elderly subjects by videofluoroscopy and manometry before and after real (19 subjects) and sham (12 subjects) exercises. A significant increase was found in the magnitude of the anterior excursion of the larynx, the maximum anteroposterior diameter, and the cross-sectional area of the UES opening after the real exercise (P < 0.05). These changes were associated with a significant decrease in the hypopharyngeal intrabolus pressure studied in 12 (real-exercise) and 6 (sham-exercise) subjects (P < 0.05). A similar effect was not found in the sham-exercise group. In normal elderly subjects, deglutitive UES opening is amenable to augmentation by exercise aimed at strengthening the UES opening muscles. This augmentation is accompanied by a significant decrease in hypopharyngeal intrabolus pressure, indicating a decrease in pharyngeal outflow resistance. This approach may be helpful in some patients with dysphagia due to disorders of deglutitive UES opening. PMID- 9227490 TI - Experimental esophagitis affects intracellular calcium stores in the cat lower esophageal sphincter. AB - We previously showed that lower esophageal spincter (LES) tone depends on spontaneous production of inositol 1,4,5-trisphosphate (IP3) and release of intracellular Ca2+ and that acute experimental esophagitis reduces LES tone and IP3 production, suggesting damage to mechanisms responsible for release of Ca2+ from intracellular stores. In the present investigation, we examined the possibility that mechanisms responsible for Ca2+ storage or uptake may also be damaged. LES circular muscle cells were isolated by enzymatic digestion. Contraction was measured in response to IP3 and thapsigargin, which enhances release of Ca2+ from intracellular stores, and in response to calmodulin and to diacylglycerol. In addition, normal cells were incubated in thapsigargin to assess the effect of depletion of intracellular Ca2+ stores on contractile response. Contraction in response to IP3 and thapsigargin was reduced in experimental esophagitis, but contraction in response to calmodulin or diacylglycerol was not. Acetylcholine (ACh)-induced contraction of normal cells was inhibited by the calmodulin antagonist CGS-9343B but not by 1-(5 isoquinolinesulfonyl)-2-methyl-piperazine dihydrochloride (H-7). In contrast, in cells from animals with esophagitis or in thapsigargin-treated cells from normal animals, ACh-induced contraction was inhibited by H-7 and not by CGS-9343B. We conclude that experimental esophagitis may damage intracellular Ca2+ stores in the LES and change the intracellular contractile pathways activated by ACh from calmodulin dependent in normal cells to protein kinase C dependent in esophagitis. PMID- 9227491 TI - GLUT2 and hexokinase control proximodistal gradient of intestinal glucose metabolism in the newborn pig. AB - We have reported previously that a high glycolytic capacity develops soon after birth in enterocytes isolated from suckling newborn pigs. In the present work, we investigated whether such metabolic changes could affect intestinal glucose utilization in vivo and examined possible variations in glucose metabolism along the small intestine. Glucose utilization by individual tissues was assessed using the 2-deoxyglucose technique. The overall glucose utilization rate was doubled in suckling vs. fasting 2-day-old pigs because of significantly higher rates in all tissues studied, except for the brain. In parallel, enterocytes were isolated from the proximal, medium, or distal jejunoileum of newborn vs. 2-day-old pigs and assessed for their capacity to utilize, transport, and phosphorylate glucose. Intestinal glucose consumption accounted for approximately 15% of glucose turnover rate in suckling vs. 8% in fasting pigs. Moreover, there was a proximal to-distal gradient of glucose utilization in the intestinal mucosa of suckling pigs. Such a gradient was also evidenced on isolated enterocytes. The stimulation of both hexokinase activity (HK2 isoform) and basolateral glucose transporter (GLUT2), as observed in the proximal jejunum, could account for such a site specific effect of suckling. PMID- 9227492 TI - Temporal expression of trefoil peptides in the TGF-alpha knockout mouse after gastric ulceration. AB - Trefoil peptides are gut peptides that have been implicated in the repair of the gastric mucosa after injury. Previous studies suggest that epidermal growth factor (EGF) receptor ligands may induce the expression of trefoil peptides. Because transforming growth factor-alpha (TGF-alpha) is a major EGF receptor ligand in the gut, we tested the hypothesis that mice with a TGF-alpha null mutation (knockout) would have reduced trefoil peptide expression compared with wild-type controls after gastric ulceration. The rate of macroscopic ulcer healing was the same in knockout and wild-type mice. Spasmolytic polypeptide (SP) and intestinal trefoil factor (ITF) expression were quantified in tissue and gastric lavage. SP and ITF levels in tissue fell within 48 h of ulceration (P < 0.05), but secretion into gastric juice was unchanged. ITF peptide expression was increased (as was SP expression) in wild-type but not knockout mice 42 and 72 days after injury (P < 0.05). The induction of SP and ITF expression in the latter stages of injury repair has a TGF-alpha-dependent component and suggests a role for these peptides in gastric differentiation and cell positioning. PMID- 9227493 TI - Effects of telenzepine and L-364,718 on canine pancreatic secretion before and after vagotomy. AB - In six conscious dogs we compared the action of the M1-receptor antagonist telenzepine (20.25-81.0 nmol.kg-1.h-1), the cholecystokinin (CCK) antagonist L 364,718 (0.025-0.1 mg.kg-1.h-1), and combinations of both on the pancreatic secretory response to intraduodenal tryptophan, given against a secretin background before and after truncal vagotomy. Before vagotomy, the higher doses of telenzepine and of L-364,718 significantly (P < 0.05) decreased the protein response to tryptophan by up to 97%. After vagotomy, all doses of L-364,718 abolished the protein response, whereas telenzepine had no further effect. Before and after vagotomy, all combinations abolished the protein response. The plasma CCK-like immunoreactivity basally, during secretin, and in response to tryptophan was not altered by vagotomy, telenzepine, and/or L-364,718. These findings indicate that in dogs 1) potentiation exists between M1 receptors and CCK for stimulation of the pancreatic enzyme response to intraduodenal tryptophan, 2) the cholinergic fibers of the enteropancreatic reflex activated by tryptophan run within the vagus nerves and end at least in part on M1 receptors, 3) CCK acts in part independently of the vagal nerves, and 4) the CCK release by intestinal tryptophan is not influenced by vagotomy, telenzepine, and/or L-364,718. PMID- 9227494 TI - Hypertrophy of colonic smooth muscle: structural remodeling, chemical composition, and force output. AB - The cellular basis of adaptations occurring during the development of megacolon was studied with the lethal spotted mouse model. Age-dependent changes in the length-force characteristics of the colon reach a steady state by 3-4 mo and include an increased relative force development at very short muscle lengths. In megacolon the following occur: 1) structural remodeling expressed as a greater increase in the fraction of maximum force production at short lengths, a shift of optimum length (Lo) to longer lengths, and no change in force per square centimeter; 2) hypertrophy and hyperplasia of both circular and longitudinal muscle; 3) high resting compliance consistent with no disproportionate change in collagen or elastin composition; 4) marked distension so that in situ circumference approximately 1.8 Lo, where active force production is low, and 5) slack length approximately 0.65 Lo, as in normal colon. Biochemical remodeling in megacolon includes disproportionate increases in ATP and phosphocreatine concentration, with 3.5-fold more preformed phosphagen than in normal colon. The myosin concentration is the same in both muscles, but the actin concentration is 1.5-fold greater in megacolon. Most of the cellular changes in megacolon would facilitate high active force output from the muscle at much larger intestinal diameters. PMID- 9227495 TI - Hypertrophy of colonic smooth muscle: contractile proteins, shortening velocity, and regulation. AB - Smooth muscle in megacolon was studied in the lethal spotted mouse and its normal sibling. In megacolon, structural remodeling and a very large increase in total protein content are associated with some changes in the contractile protein isoform composition. 1) There is a higher actin concentration in megacolon, primarily caused by a larger proportion of gamma-isoforms. 2) There was no difference in myosin concentration or in SM1/SM2 heavy chains in megacolon and normal muscle contractile proteins. 3) Only LC17a essential light chain is present in both normal and megacolon. 4) The 25- to 50-kDa 5'-insert occurs in 15 20% of the myosin in normal colon, compared with 5- to 10-fold lower amounts in megacolon. In permeabilized muscles there was no significant difference in unloaded shortening velocity (Vo) with maximal thiophosphorylation of the 20-kDa light chains, nor was there significant difference in the force vs. Ca2+ and force vs. myosin light chain phosphorylation relationships. At approximately 60% myosin light chain phosphorylation after Ca2+ activation, Vo of megacolon was approximately two times faster than Vo of normal muscle. These cellular changes largely account for the higher propulsive velocity of the colon in situ. The distribution of myosin and actin isoforms and the lack of a simple relationship between myosin light chain phosphorylation and Vo point to the operation of additional regulatory processes. PMID- 9227496 TI - Kupffer cells contain a glycine-gated chloride channel. AB - Here the effect of glycine on intracellular Ca2+ concentration ([Ca2+]i) in cultured Kupffer cells stimulated with lipopolysaccharide (LPS) was investigated to assess the possibility that they contain a glycine-gated chloride channel. LPS (10 micrograms/ml) increased [Ca2+]i rapidly, with peak values reaching 307 +/- 29 nM. Glycine (1 mM) prevented this increase nearly completely. Low concentrations of strychnine (1 microM), a glycine receptor antagonist, reversed the inhibitory effect of glycine completely; however, high concentrations of strychnine (1 mM) mimicked glycine. The effects of glycine and high-dose strychnine were prevented when cells were incubated in chloride-free buffer. Furthermore, potassium (25 mM) and LPS depolarized the Kupffer cell plasma membrane, whereas glycine caused hyperpolarization and prevented depolarization due to potassium and LPS. Moreover, tumor necrosis factor-alpha (TNF-alpha) production in cultured Kupffer cells due to LPS was decreased significantly by glycine. Therefore, it is concluded that Kupffer cells contain a glycine-gated chloride channel similar to that described previously in the central nervous system. Prevention of increases in [Ca2+]i due to LPS by activation of chloride influx reduced synthesis and release of toxic mediators by Kupffer cells. PMID- 9227497 TI - Hepatocytes in the bile duct-ligated rat express Bcl-2. AB - Hepatocytes do not express Bcl-2, a repressor of apoptosis. In contrast, cholangiocytes, which are in direct contact with bile, do express Bcl-2. Because cholestasis results in the retention of bile within hepatocytes, we reasoned cholestasis may induce hepatocellular expression of Bcl-2. Thus our aim was to determine whether hepatocytes express Bcl-2 or alter expression of other Bcl-2 family members in cholestasis using the bile duct-ligated (BDL) rat as a model of cholestasis. De novo Bcl-2 expression was observed in hepatocytes of BDL rats assessed by reverse transcriptase-polymerase chain reaction and immunoblot analysis. Immunohistochemistry demonstrated that Bcl-2 expression in hepatocytes was greater in periportal hepatocytes than pericentral hepatocytes. Expression of Bcl-x (an antiapoptotic Bcl-2 family protein) was not altered by bile duct ligation, whereas expression of Bax (a proapoptotic Bcl-2 family protein) increased slightly as determined by Northern and Western blot analyses. Bcl-2 positive hepatocytes isolated from BDL rats were resistant to induction of apoptosis by 50 microM glycochenodeoxycholate. Our results demonstrate, for the first time, expression of Bcl-2 by hepatocytes during cholestasis. We suggest that hepatocellular expression of Bcl-2 during cholestasis is an adaptive phenomenon to resist apoptosis by toxic bile salts. PMID- 9227498 TI - Epithelial localization of a reptilian Na+/H+ exchanger homologous to NHE-1. AB - Basolateral membranes of turtle (Pseudemys scripta) colon epithelial cells exhibit robust Na+/H+ exchange activity that can be activated by cell shrinkage and is blocked by amiloride [M. A. Post and D. C. Dawson. Am. J. Physiol. 262 Cell Physiol. 31):C1089-C1094, 1992]. The colonic epithelium actively absorbs Na+ and secretes K+ and HCO3-, but the role of basolateral Na+/H+ exchange, if any, in transepithelial transport is unknown. The current studies were undertaken to identify the gene product(s) responsible for the observed basolateral Na+/H+ exchange activity and to determine the cellular localization of the reptilian Na+/H+ exchange protein. We cloned and sequenced partial-length cDNAs that are likely to encode a reptilian homologue of the mammalian NHE-1 Na+/H+ exchanger isoform. The partial-length cDNAs were > 80% identical to mammalian NHE-1 homologues at the nucleotide level and recognized a transcript (approximately 5.8 6.0 kb) in RNA isolated from turtle colon, small intestine, stomach, kidney, urinary bladder, heart, and liver. In situ hybridization showed that mRNA encoding the reptile homologue of NHE-1 was expressed predominantly in the epithelial cells of these tissues. Immunofluorescent localization of the reptilian Na+/H+ exchanger protein using an antibody raised against a human NHE-1 fusion protein confirmed that protein expression paralleled abundant mRNA expression in epithelial cells of turtle stomach and colon, as well as in some nephron segments, and showed that the reptile NHE-1 homologue was localized exclusively to the basolateral membranes of these cells. The relatively high level of NHE-1 expression in epithelial cells, particularly those of the colon and stomach, suggests that NHE-1 function is important for the maintenance or regulation of ion transport processes that occur in these cell types. PMID- 9227499 TI - A novel motility effect of tachykinins in normal and inflamed colon. AB - The role of tachykinins in stimulating phasic and giant migrating contractions (GMCs) in the normal and inflamed colon in conscious dogs was investigated by close-intra-arterial infusions of test substances. At low doses (0.1 nmol), substance P and neurokinin (NK1) receptor agonist ([Sar9,Met(O2)11]substance P] stimulated phasic contractions only. At higher doses (2.0 nmol), they stimulated phasic contractions and GMCs. The phasic contractions were blocked partially but significantly by prior close-intra-arterial infusions of tetrodotoxin and atropine but not by hexamethonium. NK1 receptor antagonist partially but significantly inhibited the phasic contractile response to substance P, whereas NK2 and NK3 receptor antagonists had no significant effect. The contractile response to NK2 receptor agonist was less than one-half of the response to substance P; NK3 receptor agonist did not stimulate any contractile activity. The stimulation of GMCs by higher doses of substance P was not blocked by prior infusions of atropine, tetrodotoxin, or NK1, NK2, and NK3 receptor antagonists, nor was the contractile response to substance P blocked by H1 and H2 receptor antagonists. Inflammation depressed the phasic contractile response but enhanced the stimulation of GMCs by substance P. The ability of substance P to stimulate GMCs is novel and suggests its potential role in increasing the frequency of these contractions during colonic inflammation. PMID- 9227501 TI - Cell cycle kinetics in the alveolar epithelium. AB - The type II alveolar epithelial cell has important metabolic and biosynthetic functions but also serves as the stem cell of the alveolar epithelium. Much of the evidence underlying this premise was obtained before 1980 and provided the basis for a working model that has not been reconsidered for more than fifteen years. With the exceptions to be discussed below, little evidence has accumulated in the interim to suggest that the model requires significant alteration. Important questions remain unanswered, however, and some components of the model need to be supplemented, particularly in light of recent investigations that have provided insights not possible in earlier work. In particular, in vitro studies have suggested that the relationship between the parent type II cell and its progeny may not be as straightforward as originally thought. In addition, the rate of epithelial cell loss was recognized long ago to be an important factor in the regulation of this system, but its kinetics and mechanisms have received little attention. These and other unresolved issues are critical to our understanding of the homeostasis of the alveolar epithelium under normal and pathological conditions. PMID- 9227500 TI - Bombesin-stimulated ceramide production and MAP kinase activation in rabbit rectosigmoid smooth muscle cells. AB - We have investigated the hypotheses that 1) bombesin activation of protein kinase C (PKC) results in the hydrolysis of sphingolipids and the production of ceramide and that 2) ceramide produced on activation by bombesin mediates sustained contraction of smooth muscle cells by activation of PKC and mitogen-activated protein (MAP) kinase. Ceramide production was assessed using a technique that involved benzoylation of purified ceramide extracts, followed by reverse-phase high-performance liquid chromatography. Contraction of smooth muscle cells isolated from the rabbit rectosigmoid and stimulated with bombesin gave a significant increase in newly formed ceramide (38 +/- 3.5%). 12-O tetradecanoylphorbol-13-acetate also induced production of ceramide, which was blocked by calphostin C. The short-chain permeable C2 ceramide induced a sustained contraction and activation of MAP kinase, which was blocked by calphostin C. The increase in MAP kinase activity was maximal at 30 s and declined at 2 min. The data suggest that stimulation of smooth muscle cells by bombesin results in a functional coupling between sn-1,2-diacylglycerol (DAG)/ PKC and a sphingomyelinase, whereby DAG activates the hydrolysis of sphingomyelin to produce ceramide. Ceramide in turn activates PKC, which then activates MAP kinase. This could be the basis for the sustained contraction observed with bombesin. PMID- 9227502 TI - Adenovirus E1A upregulates interleukin-8 expression induced by endotoxin in pulmonary epithelial cells. AB - Adenovirus E1A DNA and proteins are frequently detected in lungs of patients with chronic obstructive pulmonary disease. Because adenovirus E1A can regulate host gene expression by interacting with cellular transcription factors, we postulate that E1A enhances synthesis of inflammatory mediators. To examine this possibility, we measured the expression of inflammatory cytokines in E1A producing A549 human pulmonary epithelial cells and control cells. Interleukin (IL)-8 mRNA was markedly induced by lipopolysaccharide (LPS) in E1A-producing cells but not in controls. IL-8 protein levels were elevated in parallel. In both cell types, monocyte chemotactic and activating factor mRNA induced by LPS was low, and transforming growth factor-beta 1 mRNA was not affected. IL-1 beta, IL 6, granulocyte macrophage colony-stimulating factor, and granulocyte colony stimulating factor mRNAs were too low to show any effect of E1A. We conclude that the LPS responsiveness of A549 pulmonary epithelial cells is altered by adenoviral E1A by upregulating IL-8. We speculate that this mechanism may be important in the amplification of the inflammatory process of lungs to other irritants. PMID- 9227503 TI - 5-Lipoxygenase products are necessary for ovalbumin-induced airway responsiveness in mice. AB - To determine the role of 5-lipoxygenase products in the development of airway reactivity that follows antigen exposure, we sensitized mice by intraperitoneal injection of ovalbumin and aluminum hydroxide and serial exposure to aerosols of ovalbumin. Mice lacking a functioning 5-lipoxygenase enzyme were produced by targeted gene disruption. They and their wild-type controls had measurements of lung resistance (RL) made in response to intravenous methacholine; bronchoalveolar lavage fluid cell counts and serum immunoglobulin concentrations were also measured. Wild-type mice developed striking increases in cholinergic responsiveness; 5-lipoxygenase-deficient mice manifested minimal alterations in methacholine responsiveness (RL at the highest methacholine dose was 9.9 +/- 2.4 cmH2O.ml-1.s-1 under control conditions vs. 27.6 +/- 4.6 cmH2O.ml-1.s-1 after ovalbumin in wild-type mice; 5.9 +/- 0.9 vs. 7.01 +/- 2.2 cmH2O.ml-1.s-1 in 5 lipoxygenase-deficient mice). Ovalbumin provoked airway eosinophilia and increased immunoglobulins in wild-type mice, which were present to a significantly lesser degree in 5-lipoxygenase-deficient mice. We conclude that 5 lipoxygenase products are essential for the production of nonspecific airway reactivity in mice and suggest that 5-lipoxygenase products may be important in immunoglobulin formation. PMID- 9227504 TI - Prenatal hormones alter antioxidant enzymes and lung histology in rats with congenital diaphragmatic hernia. AB - Prenatal administration of dexamethasone (Dex) and thyrotropin-releasing hormone (TRH) synergistically enhances lung maturity, but TRH suppresses the antioxidant enzyme activity. Prenatal hormonal therapy improves alveolar surfactant content and lung compliance in rats with congenital diaphragmatic hernia (CDH). In full term neonatal rats with CDH we studied the effects of prenatal Dex or Dex+TRH on antioxidant enzyme activity at birth, on survival, and on lung morphometry after 4 h of ventilation with 100% O2. CDH was induced by administration of 2,4 dichlorophenyl-p-nitro-phenylether (Nitrofen) on gestational day 10. Dex+TRH treated CDH rats had lower activity of glutathione reductase after birth than did sham-treated CDH pups. Dex-treated and sham-treated pups had similar antioxidant enzyme activity. Hormonal treatment did not change survival during ventilation. The average airspace volume increased in Dex-treated CDH pups after ventilation, with a small synergistic effect after addition of TRH. On the basis of our findings, we speculate that prenatal administration of Dex is the best choice to improve lung maturity and airspace volume in CDH patients. PMID- 9227505 TI - PAMP is a novel inhibitor of the tachykinin release in the airway of guinea pigs. AB - Proadrenomedullin NH2-terminal 20 peptide (PAMP), a newly identified hypotensive peptide, may play physiological roles in airway and cardiovascular controls. This study was designed to determine the mechanism responsible for the bronchoprotective effects of PAMP on capsaicin-induced bron-choconstriction in anesthetized guinea pigs. PAMP (10(-8)-10(-6) M) significantly inhibited capsaicin-induced bronchoconstriction in a dose-dependent manner. The bronchoprotective effect of PAMP (10(-6) M) was as large as that of isoproterenol (10(-7) M) and lasted > 10 min. The concentration of immunoreactive substance P (SP) in bronchoalveolar lavage fluid after administration of capsaicin (4 x 10( 6) M) was 120 +/- 10 fmol/ml. PAMP significantly inhibited the release of immunoreactive SP in a dose-dependent manner (60 +/- 6 fmol/ml for (10(-6) M PAMP, P < 0.01; 84 +/- 6 fmol/ml for 10(-7) M PAMP, P < 0.01; and 95 +/- 6 fmol/ml for 10(-8) M PAMP, P < 0.05). PAMP (10(-6) M) did not significantly affect exogenous neurokinin A (NKA) or NKA + SP-induced bronchoconstriction, whereas isoproterenol (10(-7) M) significantly inhibited exogenous tachykinin induced bronchoconstriction. These findings suggest that the bronchoprotective effects of PAMP are mainly due to inhibition of the release of tachykinins at airway C-fiber endings. PMID- 9227506 TI - Effect of SP-A and surfactant lipids on expression of cell surface markers in the THP-1 monocytic cell line. AB - Pulmonary surfactant and its lipid components inhibit cell proliferation and cytokine expression. Surfactant protein A (SP-A) can stimulate these same functions. We assessed the impact of SP-A and surfactant lipids on the expression of the cell surface markers, CD14, CD54 (intercellular adhesion molecule-1), and CD11b, by the human monocytic cell line THP-1 using fluorescent antibody staining and fluorescence-activated cell sorting. Under basal conditions CD14 and CD54 were undetectable, and CD11b was expressed at low levels. Incubation of the cells in 1,25(OH)2D3 alone, or with low doses of surfactant lipids, increased CD14, CD54, and CD11b. Expression was increased further by SP-A. However, the SP-A induced increases in cell markers were blocked by simultaneous treatment with lipid. The results suggest that the ability of the macrophage to participate in an inflammatory response is enhanced by SP-A alone or by surfactant containing a higher than normal proportion of SP-A. They further suggest that the addition of lipids results in a phenotype less prone to initiate an inflammatory reaction. PMID- 9227507 TI - Endothelin-1 inhibits the expression of inducible nitric oxide synthase. AB - Because nitric oxide (NO.) and endothelin (ET)-1 frequently have opposing effects on physiological and inflammatory processes, we sought to determine whether ET-1 regulates NO. synthesis by the inducible isoform of NO. synthase (iNOS). L2 cells are a rat lung epithelial cell line that synthesizes ET-1 and in which ET-1 has an autocrine role. In the current study, we demonstrate that L2 cells generate the oxidative products of NO., nitrite and nitrate, after exposure to tumor necrosis factor-alpha, lipopolysaccharide, and interferon-gamma. Exposure to these cytokines also dramatically increases the expression of iNOS mRNA. NG monomethyl-L-arginine, dexamethasone, and cycloheximide prevent the cytokine mediated increase in NO. oxidative products, demonstrating that iNOS accounts for their generation. Because L2 cells synthesize ET-1, to test the effect of removing endogenous ET-1, we used phosphoramidon (an ET-converting enzyme inhibitor) or BQ-123 (an ET receptor A antagonist). Removal of endogenous ET-1 with either phosphoramidon or BQ-123 significantly augments cytokine-stimulated NO. synthesis by approximately 20%. To further test the effect of ET-1 on iNOS, we treated cells with phosphoramidon to inhibit endogenous ET-1 synthesis and then administered ET-1 (10(-9) to 10(-7) M). In this setting, ET-1 significantly decreases inducible NO. production by 33% and iNOS mRNA by 50%. We conclude that ET-1 can decrease inducible NO. synthesis by cytokine-stimulated lung epithelial cells. PMID- 9227508 TI - Extracellular ATP stimulates K+ secretion across cultured human airway epithelium. AB - Extracellular ATP applied to the luminal side of human airway epithelium (HAE) activates an apical membrane Cl- conductance and transepithelial Cl- secretion. However, in some HAE preparations, we have found that luminal ATP induces a change in short-circuit current (Isc), consistent with K+ secretion. Using intracellular microelectrodes and radioisotopic flux studies, we investigated whether extracellular ATP regulates transepithelial K+ secretion in primary HAE cultures. In physiological Ringer solution, HAE had a negligible electrochemical driving force for Cl- secretion (DFCl), and luminal ATP induced a change in Isc opposite in polarity to Cl- secretion. Intracellular microelectrode measurements indicated that the "reversed" Isc was associated with activation of a hyperpolarizing (K+) conductance in the apical membrane. Radioisotope studies of HAE pretreated with amiloride to induce a favorable DFCl revealed that luminal ATP stimulates a small 42K secretory flux concurrently with Cl- secretion. In ion substituted Ringer solution, luminal ATP stimulated both the outward (K+) current and the inward (Cl-) current with approximately equal potency (approximately 10( 6) M). We conclude that luminal ATP activates an apical membrane K+ conductance and transepithelial K+ secretion across HAE. PMID- 9227509 TI - Nitric oxide donors inhibit spontaneous depolarizations by L-type Ca2+ currents in alveolar epithelial cells. AB - L2 cells, a cloned pneumocyte-derived cell line, express voltage-dependent L-type Ca2+ channels, causing transient depolarizing spikes of the membrane potential (Vm) [P. Dietl, T. Haller, B. Wirleitner, H. Volkl, F. Friedrich, and J. Striessing. Am. J. Physiol. 269 (Lung Cell. Mol. Physiol. 13): L873-L883, 1995]. In this study, we examined the effect of nitric oxide (NO)- and guanosine 3',5' cyclic monophosphate (cGMP)-dependent cell signaling on the activity of L-type Ca2+ channels. Using conventional microelectrodes, spontaneous depolarizations (SD) of Vm by activation of these channels are regularly seen in the presence of 10 mM bath Sr2+. The NO donors sodium nitroprusside (SNP; 1 mM), 3 morpholinosydnonimine (SIN-1; 100 microM), as well as S-nitroso-N-acetyl-D,L penicillamine (SNAP; 10 microM) caused a significant reduction of the frequency of Sr(2+)-induced SD. These effects were completely reversed by 6-anilino-5,8 quinolinequinone (10 microM), an inhibitor of the soluble guanylyl cyclase, and could be mimicked by 8-bromoguanosine 3'5'-cyclic monophosphate (8-BrcGMP; 100 microM). Perforated patch-clamp experiments revealed that 8-BrcGMP exerted a significant decrease of the depolarization-induced L-type Sr2+ current in the majority of tested cells. Consistent with the dependency of these NO-mediated effects on cGMP, incubation of L2 cells with SNP, SIN-1, and SNAP lead to a pronounced increase of cellular cGMP concentration. We conclude that the NO donors inhibit the activity of L-type Ca2+ channels in L2 cells via a cGMP dependent pathway. In the alveoli, this might occur under conditions associated with the release of NO. PMID- 9227510 TI - Nitric oxide activates chloride currents in human lung epithelial cells. AB - Epithelial Cl- channels are regulated by various physiological factors, including guanosine 3',5'-cyclic monophosphate (cGMP). Because cGMP mediates many of the physiological actions of nitric oxide (NO), we have studied both the presence of endogenous NO and the effects of exogenous NO on Cl- currents in A549 human lung epithelial cells. We have detected Ca(2+)-dependent NO synthase activity in A549 cells. Using the perforated patch-clamp technique, we have shown that inhibition of this enzyme by NG-monomethyl-L-arginine decreased Cl- current, an effect that was reversed by the NO donor S-nitrosoglutathione (GSNO). In addition, the NO donors GSNO and S-nitroso-N-acetyl-D,L-penicillamine increased whole-cell Cl- currents in A549 cells. This stimulatory effect of the NO donors was sensitive to inhibition by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, suggesting that channels other than the cystic fibrosis transmembrane conductance regulator (CFTR) are involved in the action of NO on A549 cells. In addition, 1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, a selective inhibitor of soluble guanylyl cyclase, decreased NO-mediated stimulation of Cl- currents. Our results suggest that, in lung epithelial cells, NO regulates a non-CFTR Cl- conductance acting via a cGMP-dependent mechanism. PMID- 9227511 TI - Expression of functional gap junctions in cultured pulmonary alveolar epithelial cells. AB - Recent observations suggest that cell-cell interactions may modulate the response of the alveolar epithelium to injury. Expression and function of gap junctions were thus evaluated in isolated alveolar type II cells. Freshly isolated (day 0) type II cells expressed mRNAs for gap junctional connexins 26, 32, and 43. Whereas connexin 26 mRNA declined approximately 40% in cultured cells, connexin 32 message decreased rapidly and was not detectable on day 1. In contrast, connexin 43 expression increased 10-fold by day 3 compared with day 0. Western blot confirmed a 30-fold elevation in connexin 43 protein. Connexin 45 mRNA was not detected. Functional gap junction-mediated calcium signal propagation was monitored using fura 2, a calcium-sensitive dye. Membrane deformation in a single type II cell increased intracellular calcium; the signal spread rapidly to neighboring cells by octanol-sensitive pathways. Transfer of Lucifer yellow between cells also was inhibited by octanol. These observations demonstrate functional gap junctions between cultured alveolar epithelial cells and suggest that gap junctional expression and gap junction-mediated cell-cell coupling are regulated with time in culture. PMID- 9227512 TI - Hepatocyte growth factor stimulates the differentiation of human tracheal epithelia in vitro. AB - Hepatocyte growth factor (HGF) can influence epithelial cell growth and differentiation. We examined the actions of recombinant human HGF (rhHGF) on the differentiation of human primary tracheal epithelial (HTE) cells cultured in vitro for up to 96 h. Basolateral, but not apical, treatment of confluent HTE cell sheets for 48 h with rhHGF led to increases in cell height, cell volume, cilia, and total protein content. Basolateral rhHGF treatment produced a decrease in HGF receptor (c-met) expression but had no effect on c-met mRNA levels. HTE cell sheets treated with rhHGF for 48 h showed a significant increase in mediator induced Cl- secretion and a decrease in amiloride-sensitive sodium absorption. No effect on transepithelial resistance was observed with rhHGF treatment. The enhancement of short-circuit responses by basolateral rhHGF was dose dependent. Our results demonstrate that rhHGF has hypertrophic actions on, and can influence the differentiation of, human airway epithelia in vitro, presumably through the activation of c-met at the basolateral surface of these cells. PMID- 9227513 TI - Mucus glycoconjugate secretion in cool and hypertonic solutions. AB - For sensitive individuals, exercise-induced asthma is triggered by cold and dry air and is often accompanied by a productive cough. In this study, we determined whether cold solutions and/or solutions of increased tonicity directly caused an increase in glycoconjugate (GC) secretion. To test this, we used isolated swine tracheal submucosal gland cells (TSGCs) and measured the rate of GC secretion at 37 and 32 degrees C in isotonic solutions and in solutions made hypertonic by 30 mosM. TSGCs were isolated under conditions that minimized the rate of GC secretion and were perfused with medium 199 equilibrated with 5% CO2 to a pH of 7.4. A lectin-based assay was used to specifically detect GC present in each 2 min fraction of the perfusate. Basal secretion was 3.1-fold greater at 32 degrees C (n = 3) than at 37 degrees C (n = 4; P < 0.05). At 37 degrees C, increasing perfusate osmolarity by 30 mosM increased the average rate of secretion by 41 +/- 11% (n = 4; P < 0.05); return to isotonic perfusate caused a 4.5 +/- 1.8-fold transient increase in secretion (n = 4; P < 0.05) that was complete within 10 min. At 32 degrees C, changing tonicity of the perfusate had no significant effect but returning to isotonic perfusate caused a 2.3 +/- 0.7-fold transient increase in secretion (n = 3; P < 0.05). Thus key stimuli that trigger obstruction of airflow (cold and increased osmolarity) can also directly stimulate GC secretion in the airway. Such increased secretions may contribute to the productive cough observed in some individuals in response to cold air. PMID- 9227514 TI - Atrial natriuretic peptide accounts for increased cGMP in hypoxia-induced hypertensive rat lungs. AB - Perfusate levels of nitric oxide (NO)-containing compounds and guanosine 3',5' cyclic monophosphate (cGMP) are increased in hypoxia-induced hypertensive rat lungs. To test if increased cGMP was due to NO stimulation of soluble guanylate cyclase (sGC), we examined effects of inhibition of NO synthase with N omega nitro-L-arginine (L-NNA) on perfusate accumulation of cGMP in physiological salt solution (PSS)-perfused hypertensive lungs isolated from rats exposed for 3-4 wk to hypobaric hypoxia. Because 200 microM L-NNA did not reduce cGMP, we next examined inhibitors of other pathways of stimulation of either sGC or particulate GC (pGC). Neither 5 microM Zn-protophorphyrin, an inhibitor of CO production by heme oxygenase, nor 10 mM aminotriazole, an inhibitor of H2O2 metabolism by catalase, reduced perfusate cGMP. However, an antiserum to atrial natriuretic peptide (ANP; 100 microliters antiserum/30 ml PSS), to inhibit ANP activation of pGC, completely prevented accumulation of the nucleotide. ANP antiserum was also more effective than L-NNA in reducing lung tissue cGMP. In contrast, L-NNA but not ANP antiserum increased resting vascular tone. These results suggested that whereas ANP determined perfusate and tissue levels of cGMP, NO regulated vascular tone. To test if perfusate cGMP reflected ANP stimulation of pGC in endothelial rather than smooth muscle cells, we examined effects of 10 microM Zaprinast, an inhibitor of cGMP hydrolysis in smooth muscle but not endothelial cells, and found no increase of cGMP in hypertensive lungs. ANP levels were not elevated in hypertensive lungs, and it is unclear by what mechanism the ANP-stimulated activity of pGC is increased in hypertensive pulmonary vascular endothelial cells. PMID- 9227515 TI - Nitric oxide attenuates hydrogen peroxide-mediated injury to porcine pulmonary artery endothelial cells. AB - To examine the role of nitric oxide (.NO) in vascular endothelial cell injury, cultured porcine pulmonary artery endothelial cells (PAEC) were treated with H2O2 (100-500 microM) for 30 min in the presence or absence of the .NO donors (+/-)S nitroso-N-acetylpenicillamine (SNAP) or diethylamine nitric oxide (DEANO). H2O2 caused dose-dependent PAEC cytotoxicity detected 2 h after H2O2 treatment as the release of lactate dehydrogenase. SNAP (100 microM) and DEANO (100 microM) attenuated H2O2-induced cytotoxicity if present during H2O2 treatment. In contrast, restricting treatment with .NO donors to periods before (30 min) or after (2 h) incubation with H2O2 did not prevent PAEC injury. Furthermore, the .NO synthase inhibitor NG-nitro-L-arginine methyl ester (1 mM) sensitized PAEC to H2O2-induced injury. SNAP also attenuated H2O2-induced PAEC lipid peroxidation even if restricted to periods before or after exposure to H2O2. Thus, although .NO effectively attenuated H2O2-mediated PAEC lipid peroxidation and cytotoxicity, these effects were clearly dissociated, suggesting that the antiperoxidative effects of .NO are not sufficient to account for its cytoprotective properties. PMID- 9227517 TI - Pulmonary cell kinetics and morphometry after ozone exposure: day versus night and dose response in rats. AB - Male Sprague-Dawley rats were exposed to increasing concentrations of ozone as follows: 0.12, 0.24, 0.36, 0.6, or 0.8 ppm. Controls were kept in chambers ventilated with filtered air. One-half of the animals in ozone was exposed for 12 h a day during daytime hours, and the other one-half of the animals was exposed for 12 h during nighttime. Cumulative labeling indexes were measured after 4 and 7 days in the terminal bronchioles, large intrapulmonary airways, trachea, and nasal epithelia. The penetration of the lesions from the bronchiole-alveolar junction into the alveolar zone was measured with quantitative morphometry. After 4 days of exposure, the extent of injury was dose dependent. Labeling indexes in the terminal bronchioles were 15-20% higher in animals exposed during nighttime compared with the animals exposed during daylight hours. On the other hand, depth of penetration of ozone lesions into the centriacinar region was not significantly different in animals exposed during the night compared with animals exposed during daytime. Labeling indexes in the large airways, trachea, or nasal cavity were not influenced by time of exposure. Between days 4 and 7, the lesions in the terminal bronchioles progressed only to a minimal degree (10%). It was concluded that the pattern of centriacinar tissue remodeling 1) followed a gradient based on ozone concentration and 2) was essentially complete after only 4 days of ozone exposure. Although a difference between daytime and nighttime exposure was observed, it was not considered to be large enough to invalidate conclusions drawn from studies in which animals are exposed to ozone during daylight hours. PMID- 9227516 TI - Interferon-gamma regulation of Clara cell gene expression: in vivo and in vitro. AB - This report demonstrates that Clara cell 10-kDa protein (CC10) mRNA levels are regulated by interferon-gamma (IFN-gamma). An analysis of total lung RNA from mice given IFN-gamma intratracheally showed increased levels of CC10 mRNA compared to control animals but no significant increases in surfactant proteins B and C. These results were confirmed in a Clara cell line, mtCC1-2, generated from the lungs of a transgenic mouse expressing the SV40 large T antigen under the control of a Clara cell-specific promoter. Significant increases in mtCC1-2 CC10 mRNA levels were observed in a time- and a dose-dependent manner. The expression of transacting factors hepatocyte nuclear factors 3 alpha and 3 beta (HNF-3 alpha and HNF-3 beta) were also analyzed, and a transient increase in the expression of HNF-3 beta but not HNF-3 alpha was detected. Deoxyribonuclease I footprint analysis identified a signal transducer and activator of transcription (STAT) binding site (at nucleotides -293 to -284 of CC10) adjacent to two thyroid transcription factor-1 (TTF-1) binding sites, suggesting a potential interaction between STAT1 and TTF-1. This report reinforces the hypothesis that CC10 functions as an anti-inflammatory protein and that increases in CC10 protein may provide additional protection from inflammation and disease in the lung. PMID- 9227518 TI - Regulation of nitric oxide synthesis by oxygen in vascular endothelial cells. AB - Vascular endothelial cells synthesize nitric oxide (NO) in response to agonists that elevate cytosolic free Ca2+ concentrations. Once activated, NO synthase (NOS) requires arginine, NADPH, and O2 as cosubstrates. In this study, we investigated the role of O2 in regulating endothelial NOS activity in intact bovine aortic endothelial cells by measuring the rate of nitrite (NO2-) and nitrate (NO3-) production after conversion of NO2- to S-nitrosoglutathione before analysis or after reduction of NO2- and NO3- to NO using acidic vanadium chloride. The basal rate of NO2- production was 6.5 +/- 0.8 pmol.min-1.mg protein 1. Thapsigargin (TG, 1 microM) elevated free cytosolic Ca2+ concentration and increased the rate of NO2- synthesis. At maximal concentrations of TG, the rate of stimulated NO2- production was linear for at least 20 min and was eightfold higher than the basal rate (53.5 +/- 1.8 pmol.min-1.mg protein-1). Incubation of cells in gas mixtures chosen to produce PO2 values in the physiological range led to a progressive fall in the rate of TG-stimulated NO2- production, as O2 concentrations were reduced from that of room air. The half-maximal effective concentration for NO2- production by intact cells was found to occur at 38 Torr. PO2 values higher than that of room air did not lead to a change in the rate of TG-stimulated NO2- production. To confirm that measurement of NO2- accurately reflects total NO production, both NO2- plus NO3- were measured in buffer samples from cells incubated in either room air or N2. The sum of these NO oxidation products was inhibited similarly by hypoxia. These findings suggest that O2 is an important determinant of NOS activity in hypoxic tissues or in vascular beds such as the pulmonary arterial or fetal circulation where PO2 values in the range of 40 Torr are encountered normally. PMID- 9227519 TI - Hypoxia inhibits nitric oxide synthesis in isolated rabbit lung. AB - Nitric oxide (NO.) has been proposed to modulate hypoxic vasoconstriction in the lung. The activity of nitric oxide synthase (NOS) can be inhibited by hypoxia because molecular oxygen is a necessary substrate for the enzyme. On the basis of this mechanism, we hypothesized that NOS activity has a key role in regulation of pulmonary vascular tone during hypoxia. We measured oxidation products of NO. released into the vasculature of isolated buffer-perfused rabbit lung ventilated with normoxic (21% O2), moderately hypoxic (5% O2), or anoxic (0% O2) gas using two methods. Mean PO2 in perfusate exiting the lung was 25 Torr during anoxic ventilation and 47 Torr during moderately hypoxic ventilation. We found that the amount of the NO. oxidation product nitrite released into the perfusate was suppressed significantly during ventilation with anoxic but not moderately hypoxic gas. During normoxic ventilation, nitrite release was inhibited by pretreatment with NG-monomethyl-L-arginine, a competitive inhibitor of NOS. To confirm that changes in nitrite concentration reflected changes in NO. release into the perfusate, major oxidation products of NO. (NOx) were assayed using a method for reduction of these products to NO. by vanadium(III) Cl. Release of NOx into the perfusate was suppressed by severe hypoxia (anoxic ventilation), and this effect was reversed by normoxia. Pulmonary vasoconstriction was observed during severe but not moderate hypoxia and was related inversely to the rate of nitrite release. These observations provide evidence that decreased NO. production contributes to the pulmonary vasoconstrictor response during severe hypoxia. PMID- 9227520 TI - KGF facilitates repair of radiation-induced DNA damage in alveolar epithelial cells. AB - Administration of exogenous keratinocyte growth factor (KGF) prevents or attenuates several forms of oxidant-mediated lung injury. Because DNA damage in epithelial cells is a component of radiation pneumotoxicity, we determined whether KGF ameliorated DNA strand breaks in irradiated A549 cells. Cells were exposed to 137Cs gamma rays, and DNA damage was measured by alkaline unwinding and ethidium bromide fluorescence after a 30-min recovery period. Radiation induced a dose-dependent increase in DNA strand breaks. The percentage of double stranded DNA after exposure to 30 Gy increased from 44.6 +/- 3.5% in untreated control cells to 61.6 +/- 5.0% in cells cultured with 100 ng/ml KGF for 24 h (P < 0.05). No reduction in DNA damage occurred when the cells were cultured with KGF but maintained at 0 degree C during and after irradiation. The sparing effect of KGF on radiation-induced DNA damage was blocked by aphidicolin, an inhibitor of DNA polymerases-alpha, -delta, and -epsilon and by butylphenyl dGTP, which blocks DNA polymerase-alpha strongly and polymerases-delta and -epsilon less effectively. However, dideoxythymidine triphosphate, a specific inhibitor of DNA polymerase-beta, did not abrogate the KGF effect. Thus KGF increases DNA repair capacity in irradiated pulmonary epithelial cells, an effect mediated at least in part by DNA polymerases-alpha, -delta, and -epsilon. Enhancement of DNA repair capability after cell damage may be one mechanism by which KGF is able to ameliorate oxidant-mediated alveolar epithelial injury. PMID- 9227521 TI - Epithelial cell-conditioned media inhibits degranulation of the RBL-2H3 rat mast cell line. AB - The effect of epithelial cells on mast cell responses was investigated by examination of degranulation of the rat mast cell line RBL-2H3 after overnight culture in media conditioned by the BEAS-2B human bronchial epithelial cell line [epithelial cell-conditioned media (ECM)]. These studies indicate that BEAS-2B cells secrete an inhibitor(s) of immunoglobulin E and A-23187-mediated degranulation of the RBL-2H3 cell line. The inhibitory activities of ECM are recovered after filtration through a 3-kDa cutoff filter. Pharmacological inhibition of cyclooxygenase in the BEAS-2B cells before preparation of ECM has no effect on subsequent inhibition of mast cell degranulation by ECM. However, cycloheximide treatment of the BEAS-2B cells before the conditioning process does preclude development of mast cell inhibitor activity in ECM, suggesting that this activity depends on protein synthesis. The effects of ECM on mast cell function are reversible, demonstrating that these effects do not result from overt cytotoxicity. Finally, media conditioned by primary cultures of human respiratory epithelial cells, but not fibroblasts, influence RBL-2H3 degranulation in a manner similar to ECM, suggesting that secretion of mast cell inhibitors may be somewhat unique to epithelial cells. PMID- 9227522 TI - Reduction of extracellular Na+ causes a release of Ca2+ from internal stores in airway epithelial cells. AB - Exchange of physiological salt solution with Na(+)-free solution caused an increase in intracellular Ca2+ concentration ([Ca2+]i) in 86.3% of cultured airway epithelial cells within 75 s. [Ca2+]i returned to near baseline levels within 45 s and frequently showed oscillatory increases thereafter. When extracellular Na+ concentration ([Na+]o) was reduced to 10 and 60 mM, 59.0 and 8.0% of the cells increased [Ca2+]i, respectively. Low [Na+]o-induced increase in [Ca2+]i was not blocked by amiloride, benzamil, La3+, or the absence of extracellular Ca2+. Low [Na+]o-induced [Ca2+]i increase did not occur after thapsigargin treatment. These results indicated that low [Na+]o-induced [Ca2+]i increase is due to release of Ca2+ from intracellular stores. Because mechanical stimulation of a single cell causes a Ca2+ increase among many cells (Sanderson, M. J., A. C. Charles, and E. R. Dirksen. Mechanical stimulation and intercellular communication increases intracellular Ca2+ in epithelial cells. Cell Regul. 1: 585-596, 1990.) we assayed the effect of low [Na+]o on this mechanically induced response. In low [Na+]o, mechanically induced [Ca2+]i increase in the stimulated cell was reduced; however, [Ca2+]i increase in adjacent cells was normal. We suggest that a mechanically induced Na+ conductance in the stimulated cell contributes to [Ca2+]i changes. These signaling pathways may be involved in the maintenance of periciliary ion concentrations. PMID- 9227524 TI - Cyclin D1 is required for S phase traversal in bovine tracheal myocytes. AB - We examined the role of cyclin D1 in cultured bovine tracheal myocyte mitogenesis. Immunoblots using a polyclonal antibody against cyclin D1 showed the appearance of this protein 4 h after treatment with platelet-derived growth factor (PDGF), a potent mitogen for these cells. Immunoblots utilizing an antibody against the 110-kDa retinoblastoma protein (Rb), a downstream phosphorylation target of the cyclin D1/cyclin-dependent kinase 4 (cdk4) complex, showed reduced electrophoretic mobility of this protein as early as 8 h after PDGF treatment, suggesting phosphorylation of Rb by the cyclin D1/cdk4 dimer in vivo. Epidermal growth factor (EGF), a nonmitogen, failed to induce either cyclin D1 synthesis or Rb phosphorylation. PDGF treatment of cells transiently transfected with the full-length cyclin D1 promoter subcloned into a luciferase reporter increased luciferase activity almost threefold, demonstrating transcriptional activation of the cyclin D1 promoter with mitogenic stimulation. Finally, microinjection of individual myocytes with an affinity-purified antibody against cyclin D1 reduced the percentage of cells traversing S phase after serum stimulation, as assessed by fractional bromodeoxyuridine labeling (isotype control antibody, 0.74 +/- 0.10; anti-cyclin D1, 0.22 +/- 0.04; P = 0.0001). We conclude that 1) mitogenic stimulation of cultured bovine tracheal myocytes by PDGF induces cyclin D1 transcriptional activation and protein expression, 2) cyclin D1 expression is accompanied by Rb phosphorylation, which is evidence of increased cyclin D1-associated kinase activity in vivo, and 3) microinjection of anti-cyclin D1 antibodies inhibits cellular DNA synthesis, which is evidence that cyclin D1 is required for airway smooth muscle S phase traversal. PMID- 9227523 TI - Nitric oxide production by rat alveolar macrophages can be modulated in vitro by surfactant protein A. AB - Alveolar macrophage and type II cells are known to generate nitric oxide, which is a highly reactive molecule that plays a role in host defense against pathogens, as well as tissue damage associated with inflammation in the lung. Both types of cells are known to generate the nitric oxide by inducible nitric oxide synthase (iNOS). Surfactant-associated protein A (SP-A) from various sources (human alveolar proteinosis, rat and recombinant rat) was found to upregulate nitric oxide production by alveolar macrophages in a concentration- and time-dependent manner, whereas type II cells were unresponsive to SP-A. The increase in nitric oxide production was associated with elevation in the expression of iNOS. However, only 30-50% of the cells responded by expressing iNOS, as was observed by immunofluorescence staining. The stimulatory effect of SP-A was found to be 30-50% lower than the known nitric oxide agonists interferon gamma (IFN-gamma) and lipopolysaccharide (LPS). However, addition of the cytokines interleukin-1 or granulocyte macrophage colony-stimulating factor elevated the levels of nitric oxide production to that of LPS and IFN-gamma. Special attention was given to exclude the possibility that contaminating LPS in the various SP-A species stimulated nitric oxide production by the macrophages. Our results indicate that SP-A is the agonist and not a contaminating LPS. The data presented in this report extend our knowledge regarding the nonsurfactant related functions of SP-A. PMID- 9227526 TI - Physiological assessment of complex cardiac phenotypes in genetically engineered mice. AB - The recent development of techniques for surgical manipulation and for the assessment of cardiac physiology in genetically engineered mice has allowed scientists to address some of the most fundamental questions related to congenital and acquired forms of human heart disease. This review discusses recent advances in the techniques for studying cardiac disease using the mouse as a model system. Because cardiac overload is one of the most important stimuli for development of hypertrophy and heart failure in humans, various models of cardiac pressure and volume overload, as well as myocardial ischemia, have been developed and characterized. Moreover, it is possible to reliably examine murine cardiac physiology in vivo with microtransducers, echocardiography, and other miniaturized techniques. Sophisticated methods have also been developed to enable an examination of single-cell phenotypes of isolated cardiomyocytes derived from genetically engineered mice. These physiological assessments, coupled with conventional histology and molecular markers, have allowed the characterization of several gene-targeted and transgenic mouse models of hypertrophy and dilated cardiomyopathy, as well as mouse models of cardiac developmental defects. Such mouse models of heart disease will ultimately allow the molecular dissection of the interplay between the various factors leading to heart disease, and they may serve as a guide to appropriate therapeutic strategies for human heart disease. PMID- 9227528 TI - Absence of age-related decline in total blood volume in physically active females. AB - Total blood volume (TBV) is an important determinant of cardiovascular functional capacity and the ability to maintain homeostasis during acute thermal and orthostatic stress. The primary aim of the present study was to test the hypotheses that TBV is 1) decreased with age in healthy, sedentary females, 2) not decreased with age in physically active females, and 3) directly related to maximal aerobic activity [maximal O2 consumption (VO2max)] in women of different ages and physical activity levels. An additional aim was to determine whether the use of hormone-replacement therapy in postmenopausal women is associated with altered TBV. Resting supine plasma volume (modified Evans blue dye technique) was measured in 25 postmenopausal women [12 sedentary (post-S), 13 physically active (post-PA; mean values, respectively, age 60 and 58 yr, and VO2max = 23 and 40 ml.kg-1.min-1)] and 27 premenopausal women [12 sedentary (pre-S), 15 physically active (pre-PA; mean values, respectively, age 29 and 31 yr, and VO2max = 35 and 54 ml.kg-1.min-1)]. TBV was lower in post-S (61 ml/kg) compared with pre-S (73 ml/kg) but not in post-PA (82 ml/kg) compared with pre-PA (87 ml/kg). It was also lower in post-S compared with post-PA. The lower TBV in post-S relative to pre-S and post-PA was related to lower plasma and estimated red blood cell volumes. Volumes did not differ between users and nonusers of hormone replacement. TBV was directly related to VO2max among all subjects (r = 0.65). It is concluded that 1) aging is associated with a decrease in TBV due to reductions in both plasma and erythrocyte cell volumes in healthy, sedentary females; 2) total blood, plasma, and red blood cell volumes are maintained with age in physically active females; 3) TBV is directly related to maximal aerobic capacity in females; and 4) TBV is not different in postmenopausal women using versus not using hormone-replacement therapy. PMID- 9227527 TI - Autonomic interactions for control of atrial rate are maintained after SA nodal parasympathectomy. AB - Autonomic control of atrial rate was evaluated in anesthetized dogs by electrical stimulation of stellate ganglia and/or cervical vagi before and after the intrinsic cardiac right atrial ganglionated plexus (RAGP) was injected with the nicotinic blocker hexamethonium or the membrane stabilizing chemical lidocaine, or the RAGP was surgically removed. Injections of lidocaine or hexamethonium into or surgical removal of the RAGP eliminated the bradycardia elicited by vagal stimulation without reducing the tachycardia induced by stellate stimulation. Yet, after surgical ablation of the RAGP, the tachycardia induced by sympathetic stimulation was still reduced by 94% by parasympathetic stimulation. After injections of hexamethonium or lidocaine into the RAGP were administered, the sympathetically induced tachycardia was reduced by 39 and 85%, respectively, by parasympathetic stimulation. After RAGP ablation, when atrial rate was increased by infusion of beta-adrenergic agonists, parasympathetic stimulation reduced atrial rate by 13%. Sinoatrial (SA) nodal parasympathectomy, produced by disrupting the RAGP, eliminates direct vagal control of the SA node while leaving prejunctional parasympathetic projections to sympathetic afferents innervating the SA node intact. PMID- 9227525 TI - Airway hyperreactivity produced by short-term exposure to hyperoxia in neonatal guinea pigs. AB - Airway hyperreactivity is recognized as one of the long-term sequelae of bronchopulmonary dysplasia (BPD). Due to the improved care and prognosis of very low-birth weight infants, the incidence of BPD is increasing. There are data that suggest the increased survival of premature infants may be associated with the observed increased incidence of childhood asthma. The hyperoxia received as part of the treatment of respiratory distress syndrome is believed to be partly if not completely responsible for BPD. To gain insight into the potential role that hyperoxia might play in producing airway hyperreactivity, 4-day-old guinea pig pups were exposed to 70% oxygen or air for 96 h, and airway responsiveness to acetylcholine (ACh) was assessed both 2 and 9 days after the completion of the hyperoxia exposures. Unlike ozone, the mechanism for the persistently increased airway reactivity is not related either to the inhibition of neuronal acetylcholinesterase or inhibition of the neuronal M2 muscarinic receptor. A difference in antioxidant protection did not account for the increased response of the neonatal guinea pigs compared with hyperoxia-exposed rat pups. These data support the usefulness of the neonatal guinea pig as a model to study the mechanism responsible for hyperoxia-induced airway hyperreactivity. PMID- 9227529 TI - Release of nitric oxide and prostaglandin H2 to acetylcholine in skeletal muscle venules. AB - The role of endothelium in regulating venular resistance is not well characterized. Thus we aimed to elucidate the endothelium-derived factors involved in the mediation of responses of rat gracilis muscle venules to acetylcholine (ACh) and other vasoactive agents. Changes in diameter of perfusion pressure (7.5 mmHg)- and norepinephrine (10(-6) M)-constricted venules (approximately 225 microns in diam) to cumulative doses of ACh (10(-9) to 10(-4) M) and sodium nitroprusside (SNP, 10(-9) to 10(-4) M), before and after endothelium removal or application of various inhibitors, were measured. Lower doses of ACh elicited dilations (up to 42.1 +/- 4.7%), whereas higher doses of ACh resulted in smaller dilations or even constrictions. Endothelium removal abolished both ACh-induced dilation and constriction. In the presence of indomethacin (2.8 x 10(-5) M), a cyclooxygenase blocker, or SQ-29548 (10(-6) M), a thromboxane A2-prostaglandin H2 (PGH2) receptor antagonist, higher doses of ACh caused further dilation (up to 72.7 +/- 7%) instead of constriction. Similarly, lower doses of arachidonic acid (10(-9) to 10(-6) M) elicited dilations that were diminished at higher doses. These reduced responses were, however, reversed to substantial dilation by SQ-29548. The nitric oxide (NO) synthase blocker, N omega nitro-L-arginine (L-NNA, 10(-4) M), significantly reduced the dilation to ACh (from 30.6 +/- 5.5 to 5.4 +/- 1.4% at 10(-6) M ACh). In contrast, L-NNA did not affect dilation to SNP. Thus ACh elicits the release of both NO and PGH2 from the venular endothelium. PMID- 9227530 TI - Pial arteriolar constriction to alpha 2-adrenergic agonist dexmedetomidine in the rat. AB - Dexmedetomidine (Dex) is an alpha 2-adrenergic agonist that decreases cerebral blood flow (CBF) when administered systemically. It is unclear whether cerebral vasoconstriction is mediated by a local effect on cerebral vessels or by a remote neural mechanism. In the present study, we compared the pial arteriole responses to locally and systemically administered Dex with and without local application of the specific alpha 2-adrenergic antagonist atipamezole. Six groups of male rats (n = 7 each) were anesthetized with isoflurane and prepared for measurements of small (20-39 microns), medium (40-59 microns), and large (60-79 microns) pial arteriole diameter by intravital microscopy or for regional CBF measurement by the radiolabeled-microsphere method. Local application of Dex caused dose dependent constriction that was significant starting at 10(-8) M for small and medium-sized arterioles and at 10(-7) M for large arterioles. Constriction to 10( 5) M in small [21 +/- 2% (SE)], medium (21 +/- 2%), and large (15 +/- 1%) arterioles was almost completely blocked by local application of 10(-4) M atipamezole. Intravenous administration of Dex at 1 microgram/kg decreased CBF and caused modest arteriolar constriction that began to resolve 8 min after administration. A dose of 10 micrograms/kg constricted arterioles of all sizes with constriction beginning to resolve after approximately 10 min. Local application of atipamezole (10(-4) M) slightly blunted the response to 1 micrograms/kg of intravenous Dex but did not substantially limit constriction after 10 micrograms/kg. These data demonstrate that pial arterioles are capable of substantial constriction to Dex by a local alpha 2-adrenergic mechanism. However, the inability of locally applied atipamezole to substantially inhibit the vasoconstrictor response to systemically administered Dex suggests that Dex might also cause vasoconstriction indirectly through actions at other sites in the central nervous system. PMID- 9227531 TI - Alkalemia reduces recovery from global cerebral ischemia by NMDA receptor mediated mechanism. AB - In vitro data suggest that low tissue pH reduces, whereas extracellular alkalosis potentiates, cerebral anoxic injury via excitotoxic mechanisms. We tested the hypothesis that in vivo metabolic alkalemia potentiates defects in energy metabolism after global incomplete cerebral ischemia (12 min) and reperfusion (180 min) by an N-methyl-D-aspartate (NMDA) receptor-mediated mechanism. Brain ATP, phosphocreatine, and intracellular pH (pHi) were measured by 31P magnetic resonance spectroscopy in anesthetized dogs treated with 1) preischemic intravenous carbicarb buffer (NaHCO3+Na2CO3, Carb, n = 7); 2) carbicarb infusion plus NMDA receptor antagonist MK-801 (MK-801 + Carb, n = 7); 3) an osmotically equivalent volume of 5% NaCl (NaCl, n = 8); or 4) equivalent volume of 0.9% NaCl (Sal, n = 3). Sagittal sinus pH was raised to 7.82 +/- 0.04 before and 7.65 +/- 0.03 during ischemia in Carb vs. 7.72 +/- 0.01 and 7.60 +/- 0.01 in MK-801+Carb, 7.25 +/- 0.02 and 7.15 +/- 0.03 in NaCl, and 7.31 +/- 0.00 and 7.26 +/- 0.01 in Sal, respectively. Ischemic cerebral blood flow (CBF, radiolabeled microspheres), pHi, and ATP reduction were similar among groups. By 180 min of reperfusion, recovery of ATP was greater in MK-801+Carb (104 +/- 6% of baseline), NaCl (93 +/- 6%), and Sal (94 +/- 6%) than in Carb (47 +/- 6%). Intraischemic pHi was similar among groups, and pHi recovery did not vary among groups despite differences in sagittal sinus pH. In Carb, CBF was restored but with delayed hypoperfusion. We conclude that extracellular alkalosis is deleterious to postischemic CBF and energy metabolism, acting by NMDA receptor-mediated mechanisms. PMID- 9227532 TI - Open-system kinetics of myocardial phosphoenergetics during coronary underperfusion. AB - A novel hypothesis is proposed and tested describing open-system kinetics for myocardial phosphoenergetics. The hypothesis is that during severe coronary underperfusion there is precise matching of the rates of ATP synthesis and hydrolysis, but despite the precise balance of ATP rates, there is a decrease in the concentration of ATP and an increase in the concentration of phosphocreatine (PCr) caused by the hydrolysis of AMP to adenosine. Isolated rabbit hearts were perfused using a crystalloid medium, and coronary flow was reduced by 95% from baseline for 45 min followed by reperfusion. Phosphorus nuclear magnetic resonance spectroscopy showed a rapid decrease in PCr concentration to 25% of baseline at the onset of underperfusion followed by a gradual increase in PCr to 42% of baseline, while ATP decreased continuously to 65% of baseline. The kinetics of PCr and ATP could only be described by the precise matching of the rates of ATP synthesis and ATP hydrolysis and an open adenylate system that included the decrease in cytosolic AMP concentration via the production and efflux of adenosine. To confirm the hypothesis of open-system kinetics, two independent predictions were tested in separate experiments: 1) total coronary venous purine efflux (adenosine+inosine+hypoxanthine) during underperfusion was equal to the decrease in ATP concentration, and 2) there was no increase in PCr during moderate coronary underperfusion (80% flow reduction). In conclusion, the open nature of the myocardial adenylate system causes mass action effects that exert novel control over PCr and ATP concentrations during coronary underperfusion. The open-system kinetics cause ATP to decrease and PCr to increase, even though there is precise matching of the rates of ATP synthesis and hydrolysis. Finally, the hydrolysis of AMP to adenosine may benefit tissue survival during ischemia by improving the free energy of ATP hydrolysis, thereby delaying or preventing calcium overload. PMID- 9227533 TI - Influence of gender on vasomotor effects of oxidized low-density lipoprotein in porcine coronary arteries. AB - This study compared 5-hydroxytryptamine (5-HT)-induced contraction and relaxation in coronary arteries from male and female pigs and compared the vasomotor effects of the atherosclerotic lipoprotein, oxidized low-density lipoprotein (LDL), in these tissues. 5-HT-induced contraction and endothelium-dependent relaxation were similar, as was sodium nitroprusside-induced relaxation, in coronary arteries from male and female pigs. These data suggest that there were no gender-related differences in 5-HT-induced contraction or 5-HT-mediated nitric oxide (NO) release from the coronary endothelium. In contrast, oxidized LDL (100 micrograms/ml) enhanced 5-HT-induced contraction to a greater extent in coronary arteries from male versus female pigs. Because oxidized LDL inhibited 5-HT induced relaxation similarly in arteries from male and female animals, a greater effect of oxidized LDL on agonist-induced NO release in tissues from male pigs cannot explain the greater effect on 5-HT-induced contraction. Oxidized LDL contracted coronary arteries from males with a greater force than arteries from females when measured from baseline tone, suggesting that oxidized LDL inhibited basal NO release to a greater extent in coronary arteries from male pigs compared with females, an effect that may have participated in the greater enhancement of 5-HT-induced contraction that occurred in arteries from male pigs. These gender related differences in the vasomotor effects of oxidized LDL may play an important role in the lesser incidence of cardiovascular disease in premenopausal females than in males and may provide insight into the cardioprotective effect of estrogen. PMID- 9227534 TI - Lysophosphatidylcholine stimulates leukocyte rolling and adherence in rat mesenteric microvasculature. AB - Synthetic exogenous lysophosphatidylcholine (LPC) was used to induce leukocyte endothelial cell interaction, utilizing intravital microscopy in the rat mesenteric microvasculature. Superfusion of the rat mesentery with 10 microM LPC significantly and time dependently increased leukocyte rolling and adherence compared with control rats. Moreover, LPC superfusion did not alter systemic blood pressure or mesenteric venular shear rate. Administration of either an anti P-selectin monoclonal antibody (PB1.3 at 1 mg/kg i.v.) or a nitric oxide (NO) donor (CAS-1609, 10 microM superfusion) markedly attenuated LPC-induced leukocyte rolling and adherence (P < 0.01 from LPC alone). Immunohistochemical localization of P-selectin and intercellular adhesion molecule-1 (ICAM-1) expression on mesenteric venules was significantly increased after exposure to LPC compared with mesenteries superfused with phosphatidylcholine (P < 0.001). Flow cytometric analysis indicated that 10 microM LPC significantly (P < 0.05) increased P selectin fluorescence on rat platelets. Furthermore, direct measurement of NO release from rat aortic vascular endothelium was significantly (P < 0.05) inhibited by 10 microM LPC. Thus LPC induces in vivo leukocyte rolling and adherence in the mesenteric rat microvasculature by increasing the surface expression of P-selectin and ICAM-1. Because inhibition of endothelial NO release promotes P-selectin expression, LPC-induced rolling and adherence may be mediated via reduced NO release. PMID- 9227535 TI - Involvement of ceramide in inhibitory effect of IL-1 beta on L-type Ca2+ current in adult rat ventricular myocytes. AB - Interleukin (IL)-1 beta has previously been shown to decrease the L-type Ca2+ channel current (ICa,L). Because ceramide has been suggested to mediate many biological effects of IL-1 beta, we examined whether ceramide was involved in the IL-1 beta-induced suppression of ICa,L in adult rat ventricular myocytes. Exposure of myocytes to 5 ng/ml IL-1 beta elicited a 140% increase in ceramide production within 2 min, as measured with 32P phosphorylation. Whole cell patch clamp techniques were used to measure ICa,L in myocytes internally dialyzed and externally perfused with Na(+)- and K(+)-free solutions. C2 ceramide (1 nM-1 microM), a membrane-permeable analog of ceramide, caused a concentration dependent inhibition of ICa,L and increased the rate of ICa,L inactivation without altering its gating properties. An inactive ceramide analog failed to inhibit ICa,L. At submaximal concentrations, effects of C2 ceramide and IL-1 beta on ICa,L were additive and saturable. In the presence of a maximally effective concentration of IL-1 beta, C2 ceramide had no further effect on ICa,L. These results suggest that ceramide mediates IL-1 beta-induced suppression of cardiac ICa,L. PMID- 9227536 TI - IGF-I regulates K(+)-channel expression of cultured neonatal rat ventricular myocytes. AB - Insulin-like growth factor (IGF) I has been known as an important peptide during heart development and myocardial hypertrophy. In the present study, the effects of IGF-I on cardiac K(+)-channel expression were investigated in cultured neonatal rat ventricular myocytes. Two distinct 4-aminopyridine (AP)-sensitive and rapidly activating outward K+ currents (IK) were observed. The predominant K(+)-channel current in cultured cells was a fast inactivating current similar to a 4-AP-sensitive transient outward current [I(to) half-maximal inhibitory concentration (IC60) = 0.87 mM]. Some cells lacking I(to) expressed an IK with little or no slow inactivation. IK exhibited higher sensitivity to inhibition by 4-AP (IC50 = 66.5 microM) and could be enhanced by isoproterenol but unaffected by tetraethylammonium. These characteristics indicate that IK might be the rat isoform of ultrarapid delayed rectifier K+ current IKur previously described in human atrial myocytes. Seventy-two-hour exposure to 60 ng/ml IGF-I induced myocyte hypertrophy and increased the percentage of cells expressing only IKur and the cells expressing both Ito and IKur. In some cultured myocytes, immunofluorescent staining with a polyclonal antibody specific to the COOH terminus of Kv1.5 K(+)-channel protein was performed in the same single cells after voltage-clamp recordings. The IGF-I-pretreated cells expressing larger IKur revealed a significantly intense antibody labeling. These observations indicate that the long-term administration of IGF-I can regulate the K(+)-channel expression of cultured neonatal rat ventricular myocytes. This is important for understanding the role of IGF-I in the modulation of cardiac excitability. PMID- 9227537 TI - Glibenclamide-induced blockade of ischemic preconditioning is time dependent in intact rat heart. AB - The ATP-sensitive potassium (KATP) channel has been demonstrated to be a potential mediator of ischemic preconditioning (PC) in most species, with the exception of the rat. This conclusion is based on the failure of the KATP channel antagonist glibenclamide (Glib) to block PC in this species. However, previous studies did not take into consideration the kinetic properties of Glib binding to its receptor in the rat myocardium. Therefore, the purpose of the present study was to test the hypothesis that ischemic PC is mediated by the myocardial KATP channel in the rat and that blockade of PC by Glib is a time-dependent phenomenon. Barbiturate-anesthetized, open-chest male Wistar rats were subjected to 30 min of occlusion and 2 h of reperfusion. Ischemic PC was elicited by three 5-min occlusion periods interspersed with 5 min of reperfusion. Infarct size (IS) as a percentage of the area at risk (AAR; IS/AAR) was determined with triphenyltetrazolium staining. PC and the KATP channel opener nicorandil (50 micrograms.kg-1.min-1 i.v.) resulted in marked reductions in IS/AAR from 53 +/- 3% to 8 +/- 1% and 17 +/- 5%, respectively (P < 0.05). Two doses of Glib (1 or 0.3 mg/kg i.v.) given 30 min before PC abolished the protective effect of PC; however, Glib (0.3 mg/kg i.v.) given 5 min before PC failed to block the effect. Glib administered 30 min before the 15-min nicorandil infusion blocked the cardioprotective effect of NC but did not block its transient hypotensive effect. These results demonstrate that the KATP channel is involved in ischemic PC in the intact rat heart and that the inhibitory effect of Glib to block PC is time dependent. PMID- 9227539 TI - CD18-dependent leukocyte adherence and vascular injury in pig cerebral circulation after ischemia. AB - Recent accumulating evidence indicates that leukocytes contribute importantly to ischemic brain injury. Although large numbers of leukocytes are present in the ischemic territory of reperfused brain 24-48 h after the ischemic insult, little is known of the acute inflammatory response to cerebral ischemia, particularly regarding the time course and magnitude of leukocyte adherence to cerebrovascular endothelium and the functional consequences of such adherence. To study these issues, we developed an epifluorescence videomicroscopy system for observing and quantifying the dynamic behavior of rhodamine-labeled leukocytes in the cerebrovascular microcirculation. Anesthetized piglets equipped with closed cranial windows were used in these investigations. During the initial 2 h of reperfusion after 9 min of asphyxia (n = 6), a marked, progressive increase in adherent leukocytes was noted in cerebral postcapillary venules that was significantly greater in magnitude than that seen in nonasphyxic, time-matched controls (n = 8). A similar response was observed after complete global ischemia of 10 min duration. A significant increase in sodium fluorescein permeability was also measured at 2 h of reperfusion in asphyxic animals. Pretreating a separate asphyxic animal group (n = 7) with a monoclonal antibody to the leukocyte adhesion glycoprotein complex CD11/CD18 severely attenuated both leukocyte adherence and the increase in vascular permeability. These results provide evidence that adherent leukocytes contribute to disruption of endothelial integrity during early reperfusion after global ischemic insults, the inhibition of which may reduce the vasogenic edema that occurs early during reperfusion after birth asphyxia, stroke, and cardiac arrest. PMID- 9227538 TI - Possible role of T-type Ca2+ channels in L-NNA vasoconstriction of hypertensive rat lungs. AB - Acute inhibition of endothelium-derived nitric oxide (NO) synthesis by L-arginine analogs such as N omega-nitro-L-arginine (L-NNA) has little effect on basal vascular tone in normal rat lungs but elicits marked vasoconstriction in hypertensive lungs. The NO-suppressible vasoconstriction is dependent on extracellular Ca2+ but is not mediated by L-type Ca2+ channels. This study tested whether the response was mediated by Ca2+ influx through receptor-operated channels, reverse Na+/Ca2+ exchange, or low-threshold voltage-gated (T-type) Ca2+ channels. We first examined whether SKF-96365, a blocker of receptor-operated Ca2+ channels, inhibited L-NNA-induced vasoconstriction in salt solution-perfused hypertensive lungs isolated from chronically hypoxic male rats (exposed to hypobaria of 410 mmHg for 3-5 wk). Whereas 50 microM SKF-96365 inhibited pressor responses to angiotensin II and acute hypoxia, it did not reduce vasoconstriction in response to 100 microM L-NNA. We next examined effects of pretreatment with Na+/Ca2+ exchange blockers and observed that L-NNA vasoconstriction was reduced by both 100 microM amiloride and 50 microM ethylisopropyl amiloride (EIPA). The third experiment showed that each of two different blockers of T-type Ca2+ channels, 10 microM Ro-40-5967 and 300 microM nordihydroguariaretic acid, inhibited L-NNA vasoconstriction and that the combination of EIPA and Ro-40-5967 did not cause more inhibition than did Ro-40-5967 alone. These results suggest that, whereas receptor-operated Ca2+ channels are not significantly involved in the mechanism of NO-suppressible vasoconstriction in hypertensive rat lungs, Ca2+ influx through reverse Na+/Ca2+ exchange and/or T-type Ca2+ channels may play a role. Because both amiloride and EIPA also inhibit T-type Ca2+ channels, we speculate that Ca2+ influx through these channels rather than through reverse Na+/Ca2+ exchange is an important mediator of the vasoconstriction. PMID- 9227540 TI - Autonomic reactivity and hormonal secretion in lactate-induced panic attacks. AB - To compare autonomic and neuroendocrine responses during lactate-induced panic attacks, heart rate variability and cortisol and atrial natriuretic hormone (ANH) levels were measured in patients with panic attacks and in healthy control subjects. In a randomized double-blind design, all subjects received either 10 ml/kg body weight of 0.5 M racemic sodium lactate or normal saline from 1100 to 1120. Spectral analysis of the R-R interval of analog electrocardiograms was performed, and total (0.001-0.45 Hz), low-frequency (0.01-0.05 Hz), midfrequency (0.05-0.15 Hz), and high-frequency power (0.15-0.45 Hz) were computed. Cortisol was measured 12 times in the period from 0900 to 1300, and ANH was measured at 1100, 1120, and 1200 by radioimmunoassay. In both panickers (n = 6) and nonpanickers (n = 8), an infusion of lactate resulted in an acceleration of heart rate, a reduction in total spectral power, and a decrease in the high- and low frequency components of spectral power. Panickers showed a significant enhancement of the high-frequency power, whereas in nonpanickers, a shift from the mid- and high-frequency toward the low-frequency power emerged. ANH plasma concentrations during lactate infusion in panickers showed a significant increase (115 and 131% at 1120 and 1200, respectively, over concentrations at 1100) in contrast to nonpanickers (20 and 74%, respectively). No group or treatment effects on cortisol secretion emerged, which is in line with former reports. Our study supports preliminary observations that lactate-induced panic attacks enhance the release of ANH, a vasodilatator and inhibitor of sympathetic activity. Hence this hormone not only could inhibit the secretion of the stress hormone cortisol but, in parallel, could also attenuate the sympathetic stimulation to the heart. These inhibitory effects of ANH could explain the so far-unresolved dissociation between psychopathological alterations and autonomic and endocrine responses of panic attacks. PMID- 9227541 TI - Upregulation of expression of sarcoplasmic reticulum by TGF-beta 1 in cultured rat cardiac myocytes. AB - The effects of transforming growth factor-beta 1 (TGF-beta 1) on the function and structure of sarcoplasmic reticulum (SR) were studied in cultured neonatal rat cardiac myocytes. The cardiac myocytes at days 2 and 6 of culture exhibited spontaneous contraction; however, the rate of contraction increased and became regular, depending on culture day. Ryanodine and norepinephrine (NE) increased the rate of contraction and frequency of Ca2+ oscillations in myocytes at day 2 of culture. Ryanodine did not affect the spontaneous contraction in nontreated and TGF-beta 1-treated myocytes. On the other hand, NE caused negative and positive chronotropic responses in nontreated and TGF-beta 1-treated cells, respectively. In the absence of extracellular Ca2+, ryanodine and NE did not affect the cytoplasmic Ca2+ concentration ([Ca2+]i) in the nontreated cells, whereas NE increased [Ca2+]i but ryanodine did not in the TGF-beta 1-treated cells. SR structures in TGF-beta 1-treated cells developed more than those in nontreated cells. The results indicate that TGF-beta 1 plays an important role in the upregulation of SR function and structure of cultured neonatal rat cardiac myocytes. PMID- 9227542 TI - Ca2+ release mechanism of primate drug-induced coronary vasospasm. AB - Cellular mechanisms of protection against drug-stimulated coronary vasospasm were studied by multiweek estrogen plus progesterone (P) vs. medroxy-progesterone acetate (MPA) treatments by measuring intracellular Ca2+ and protein kinase C (PKC) signals. Ovariectomized monkeys (OVX) were treated by slow-release implants with either P or MPA for 4 wk added to estradiol-17 beta (E2) begun 2 wk earlier. A third group received E2 for 2 wk and withdrawal of E2 (W; no steroid treatment) during the last 4 wk. OVX coronary artery vascular muscle cells (VMC) in primary culture conditions were labeled by the fluorescent indicators, fluo 3 and hypericin, respectively, to study intracellular Ca2+ and PKC mechanisms of coronary artery hyperre-activity, using digital analysis of single VMC by photon counting camera. Stimulation by 10 microM serotonin and 100 nM U-46619 (thromboxane A2 mimetic) caused Ca2+ increases (2-5 min) and no PKC activation in VMC from five P-treated monkeys but prolonged (> or = 30 min) increases in both Ca2+ and PKC signals in VMC from six MPA-treated monkeys or seven W-treated monkeys; these P vs. MPA (or W) differences were maintained > or = 14 days. We hypothesize that hyperreactivity in VMC from MPA- or W-treated monkeys results from accelerated prolonged Ca2+ release, with concomitant PKC activation, and that MPA (but not P) negates the coronary vasospasm protective effect of E2. PMID- 9227543 TI - Hypoxia and the cardiovascular response to dynamic knee-extensor exercise. AB - Hypoxia affects O2 transport and aerobic exercise capacity. In two previous studies, conflicting results have been reported regarding whether O2 delivery to the muscle is increased with hypoxia or whether there is a more efficient O2 extraction to allow for compensation of the decreased O2 availability at submaximal and maximal exercise. To reconcile this discrepancy, we measured limb blood flow (LBF), cardiac output, and O2 uptake during two-legged knee-extensor exercise in eight healthy young men. They completed studies at rest, at two submaximal workloads, and at peak effort under normoxia (inspired O2 fraction 0.21) and two levels of hypoxia (inspired O2 fractions 0.16 and 0.11). During submaximal exercise, LBF increased in hypoxia and compensated for the decrement in arterial O2 content. At peak effort, however, our subjects did not achieve a higher cardiac output or LBF. Thus O2 delivery was not maintained and peak power output and leg O2 uptake were reduced proportionately. These data are consistent then with the findings of an increased LBF to compensate for hypoxemia at submaximal exercise, but no such increase occurs at peak effort despite substantial cardiac capacity for an elevation in LBF. PMID- 9227544 TI - Hypoxic pHi and function modulation by Na+/H+ exchange and alpha-adrenoreceptor inhibition in heart in vivo. AB - Regulation of intracellular pH (pHi) may contribute to maintenance of cardiac contractile function during graded hypoxia in vivo. To test this hypothesis, we disturbed pHi regulation in vivo using two approaches: alpha-adrenoreceptor antagonism with phentolamine (1 mg/kg) (Phen; n = 9); and Na+/H+ exchange inhibition with HOE-642 (2 mg/kg; n = 6) before graded hypoxia in open-chest sheep. Hemodynamic parameters including left ventricular maximal pressure development (dP/dtmax) cardiac index (CI), and left ventricular power were monitored continuously and simultaneously with high-energy phosphate levels and pHi, measured with 31P nuclear magnetic resonance spectroscopy in Phen, HOE-642, and control (Con; n = 9). In subgroups (n = 6) in Con and Phen, coronary flow, myocardial oxygen consumption (MVO2), and lactate uptake were also measured. During hypoxia, the functional parameters left ventricular dP/dtmax, CI, and left ventricular power decreased significantly compared with baseline and Con values. These decreases were preceded by a significant drop (P < 0.05) in pHi from 7.10 +/- 0.04 to 6.69 +/- 0.05 in Phen and corresponded temporally to a pHi drop from 7.10 +/- 0.02 to 6.77 +/- 0.03 in HOE-642. Decreases in pHi in Phen were not preceded by decreases in cardiac function or MVO2. In contrast, cardiac function parameters increased significantly in Con, whereas no significant pHi decrease occurred (7.07 +/- 0.03 to 6.98 +/- 0.04). We conclude that these data indicate that pHi regulation can be disrupted through alpha-adrenergic antagonism or Na+/H(+)-exchange inhibition in vivo. These studies demonstrate that pHi regulation performs a role in the modulation of cardiac function during hypoxia in vivo. PMID- 9227545 TI - New mechanoenergetic evaluation of left ventricular contractility in in situ rat hearts. AB - We recorded a series of ejecting left ventricular (LV) pressure (P)-volume (V) loops of in situ rat hearts during a gradual ascending aortic occlusion. The end systolic (ES) P-V relationship (ESPVR) was upward convex curvilinear regardless of LV contractility. The ESPVR was shifted upward in an enhanced contractility by dobutamine and downward in a depressed contractility by propranolol; ESP at a midrange V of 0.1 ml/g LV on each ESPVR increased from 131 +/- 11 to 192 +/- 17 mmHg and decreased from 136 +/- 10 to 110 +/- 7 mmHg, respectively. Furthermore, we obtained an upward concave curvilinear pressure-volume area (PVA; a measure of total mechanical energy)-V (preload) relationship to assess LV work capability in each contractility. This relationship also shifted upward in enhanced contractility and downward in depressed contractility; the PVA at midrange V increased from 7.9 +/- 1.2 to 12.3 +/- 1.5 mmHg. ml.beat-1.g-1 and decreased from 8.2 +/- 0.9 to 6.4 +/- 0.8 mmHg.ml.beat-1.g-1. We conclude that the heights of the ESPVR and PVA-V relationship curves can evaluate LV contractility mechanoenergetically. PMID- 9227546 TI - Cytosolic [Ca2+], [Na+], and pH in guinea pig ventricular myocytes exposed to anoxia and reoxygenation. AB - The aim of this study was to evaluate whether the magnitude and time course of the intracellular acidification observed in anoxic cardiac myocytes was sufficient to protect against reoxygenation-induced hypercontracture. Cytosolic [Ca2+], [Na+], and pH were measured using fluorescent indicators in myocytes that were first subjected to both anoxia and glucose deprivation and then oxygen and glucose restoration 15-30 min after the onset of rigor. The cytosol underwent a profound acidification early in anoxia (pH 7.21 to 6.84) that reached a plateau at the time of rigor contracture. In contrast, [Na+] rose throughout anoxia. Cytosolic [Ca2+] underwent little rise during anoxia, but reoxygenation induced a large spike in [Ca2+]. Reoxygenation also induced a significant secondary acidification of the cytosol that was apparently induced by the spike in [Ca2+]. The characteristics of this secondary acidification were deemed sufficient to provide partial protection against the hypercontracture associated with the reoxygenation-induced [Ca2+] transient. PMID- 9227547 TI - Activation of tyrosine kinases in H2O2-induced contraction in pulmonary artery. AB - Hydrogen peroxide (H2O2) is an important reactive oxygen species implicated in lung vascular constriction and injury. The purpose of this study was to investigate the role of tyrosine kinases in H2O2-induced vascular contraction and dysfunction. In our study, H2O2 (200 microM) caused an initial transient contraction followed by a strong, sustained contraction in isolated rat pulmonary arteries. Genistein, a tyrosine kinase inhibitor, attenuated both the initial and the sustained contractions. Aminogenistein and tyrphostin 51, specific inhibitors of tyrosine kinases, had the same effects as genistein. Exposure of pulmonary arteries to H2O2 for 1 h caused a significant reduction in the contractile response to KCl or phenylephrine and in the vasodilatory response to acetylcholine (smooth muscle dysfunction). Although tyrosine kinase inhibitors significantly blocked contractions induced by H2O2, pretreatment of pulmonary arteries with these inhibitors before H2O2 exposure did not prevent the decreases in responses to KCl, phenylephrine, or acetylcholine. Removal of extracellular Ca2+ and depletion of intracellular Ca2+ pools by ryanodine or thapsigargin did not inhibit the initial and sustained contractions in response to H2O2. W-7, a calmodulin antagonist, or ML-9, a myosin light chain kinase inhibitor, significantly inhibited the sustained contractions but did not prevent smooth muscle dysfunction induced by H2O2. These data show that 1) exposure to H2O2 causes smooth muscle contractions and dysfunction in isolated pulmonary arteries and 2) activation of tyrosine kinases mediates H2O2-induced contractions; however, tyrosine kinases do not appear to be involved in H2O2-induced inhibition of arterial responses to vasoactive substances. These data suggest that different signaling pathways and mechanisms are involved in H2O2-induced smooth muscle contraction and dysfunction. PMID- 9227548 TI - Direct coupling between blood flow and metabolism at the capillary level in striated muscle. AB - In hamster cremaster muscle, capillary networks consist of anatomically invariant subunits termed modules [Berg, B. R., and I. H. Sarelius, Am. J. Physiol. 268 (Heart Circ. Physiol. 37): H1215-H1222, 1995]. To explore local coupling between blood flow and metabolism, we used micropipettes to stimulate five to six muscle fibers running underneath specified capillary modules. Capillary erythrocyte flow increased significantly at all stimulation frequencies because of increased erythrocyte content at 2 Hz and increased erythrocyte velocity at 4 and 8 Hz. Erythrocyte flow did not increase when the fibers underlying the module were mechanically tugged but did not actively contract at these frequencies. Increased capillary flow was accommodated by dilation of three upstream arteriolar generations: the module inflow arteriole dilated significantly at all frequencies, and further upstream, dilations were significant at higher frequencies. Other module inflow arterioles in the same capillary network as the stimulated module did not dilate. Dilations in the module inflow arteriole were abolished by 600 mosM sucrose but were unaffected by 10(-6) M tetrodotoxin. These data suggest that local coupling between capillary flow and muscle contraction includes a conducted vasodilation that is responsible for the remote upstream dilations. PMID- 9227549 TI - Hypercholesterolemia is associated with enhanced angiotensin AT1-receptor expression. AB - Low-density lipoprotein increases the AT1-receptor gene expression in vascular smooth muscle cells. To elucidate whether elevated cholesterol serum levels upregulate the AT1 receptor and its functional response to angiotensin II in vivo, we compared 1) the vasoconstrictive effect of angiotensin II and 2) the level of expression of the vascular AT1 receptor in aortas of normocholesterolemic and hypercholesterolemic rabbits. Contraction experiments on isolated aortic rings showed that the angiotensin II-induced vasoconstriction was increased in hypercholesterolemic New Zealand White rabbits compared with normocholesterolemic New Zealand White rabbits. This difference in the angiotensin II-induced vasoconstriction was caused by a twofold increase in the density of cell surface AT1 receptors in hypercholesterolemic rabbits, as assessed by radioligand binding assays. The enhanced expression of AT1 receptors on the surface of these vascular cells was caused by elevated steady-state levels of the AT1-receptor mRNA to 220 +/- 35% in aortas excised from hypercholesterolemic rabbits compared with levels in aortas from normocholesterolemic rabbits, as measured by Northern blot analysis. These data indicate that hypercholesterolemia is associated with upregulation of expression and function of vascular AT1 receptors in vivo. This suggests a novel mechanism by which hypercholesterolemia could be involved in the onset and progression of chronic vascular diseases such as hypertension and arteriosclerosis if the phenomenon is confirmed in humans. PMID- 9227550 TI - LPS-induced delayed myocardial adaptation enhances acute preconditioning to optimize postischemic cardiac function. AB - Myocardial tolerance to ischemia and reperfusion (I/R) injury can be achieved by either acute or delayed cardioprotective mechanisms. Ischemic preconditioning has been demonstrated to be a powerful acute cardioprotective stimulus. We have reported that lipopolysaccharide (LPS) pretreatment induces delayed myocardial adaptation to I/R injury. To optimize myocardial protection, we examined the ability of delayed myocardial adaptation to enhance acute ischemic preconditioning in the isolated working rat heart. Male Sprague-Dawley rats were divided into control, acute [transient ischemia (TI); 5-min global ischemia, 37 degrees C], delayed (LPS; 500 micrograms/kg i.p.), or combined (LPS + TI) cardioprotective groups. Delayed cardioprotection involved LPS injection 72 h before heart isolation. All hearts were subjected to 20-min global ischemia (37 degrees C) and 30-min reperfusion. Coronary effluent collected during reperfusion was assayed for creatine kinase (CK) activity. Both TI and LPS treatment improved postischemic aortic flow recovery (29 +/- 4.5 and 44 +/- 4.0%, respectively; P < 0.05, LPS vs. TI) compared with control hearts (11 +/- 2.2%; P < 0.05, TI or LPS vs. control). When TI was applied to LPS-treated hearts (LPS + TI), aortic flow recovery was further enhanced (57 +/- 3.8%; P < 0.05 vs. TI or LPS alone). CK release during 20 and 30 min of reperfusion was decreased in all treated hearts compared with control hearts (P < 0.05). These results indicate that delayed myocardial adaptation and acute ischemic preconditioning independently activate protective mechanisms against ischemia. Enhanced protection occurs when induced delayed mechanisms are combined with acute cardioprotective stimuli, which optimize postischemic myocardial function and reduce myocellular necrosis. PMID- 9227551 TI - Diameter variability and microvascular flow resistance. AB - Microvessels are known to exhibit irregular shapes, deviating substantially from an idealized cylindrical tube geometry. Such irregularities must be taken into account in calculating microvascular flow resistance and may add to the observation that flow resistance in living microvessels in vivo is about twice that predicted on the basis of tube flow studies in vitro. The present study was aimed at providing a comprehensive database describing the apparent diameter variability for all segments of a complete microvascular network in the rat mesentery and assessing the impact of this variability on segmental flow resistance and the pressure drop across the network. Diameters were estimated by intravital microscopy at axial intervals of 20 microns along the 546 vessel segments of a mesenteric microvessel network, resulting in 6,319 separate diameter measurements. The amplitude of diameter variations in individual vessel segments decreased from approximately 15% of the mean vessel diameter in the smallest segments (approximately 5 microns diam) to approximately 5% in the largest segments (approximately 60 microns diam). Segmental hindrance was estimated to be 10-23% higher than calculated from arithmetic mean diameter, depending on the model used to estimate the hydrodynamically effective segment diameter. The overall pressure drop across the network calculated using a mathematical flow simulation was increased by 7-13.5%. This increase in flow resistance can explain approximately 10% of the observed discrepancy between flow resistance in vivo and in vitro. PMID- 9227552 TI - beta 1-and beta 2-adrenergic receptors exhibit differing susceptibility to muscarinic accentuated antagonism. AB - Neonatal rat ventricular myocytes express both beta 1-and beta 2-adrenergic receptors linked to enhanced intracellular adenosine 3',5'-cyclic monophosphate (cAMP) accumulation and the modulation of contractile function. This study tests the hypothesis that muscarinic agonists act via distinct mechanisms to interfere with beta 1-and beta 2-adrenergic receptor actions. The beta 2-selective agonist zinterol (10(-7) M) elicits approximately a fourfold increase in cAMP accumulation, which is mimicked, both in magnitude and kinetics, by 10(-9) M of the mixed beta 1-receptor agonist/beta 2-receptor agonist isoproterenol. At these concentrations, isoproterenol and zinterol elicit equivalent inotropic and lusitropic (i.e., enhanced relaxation) responses. Carbachol inhibits all three responses (cAMP, inotropic, and lusitropic) elicited by isoproterenol. In contrast, carbachol does not interfere with the effect of zinterol to augment cAMP accumulation or to induce a positive inotropic response. However, carbachol inhibits the lusitropic response to zinterol via an action at an M2-muscarinic receptor linked to a pertussis toxin-sensitive pathway. Additional studies indicate that beta 2-receptor-dependent phosphorylation of troponin I and phospholamban is substantially attenuated by carbachol. We conclude that carbachol interferes with beta 1-receptor actions by reducing cAMP accumulation. In contrast, the anti-beta 2-receptor actions of carbachol are mediated by a mechanism that is distinct from inhibition of cAMP accumulation, involving an M2 muscarinic receptor coupled to a pertussis toxin-sensitive G protein, which leads to inhibition of troponin I and phospholamban phosphorylation and inhibition of the beta 2-receptor-dependent lusitropic response. PMID- 9227554 TI - VEGF effect on vascular permeability is mediated by synthesis of platelet activating factor. AB - Vascular endothelial growth factor (VEGF), a protein synthesized and secreted by a variety of normal and neoplastic cells, serves as an endothelial cell-specific mitogen and a potent angiogenic factor. Intradermal injection of VEGF increases vascular permeability, a critical event in inflammation and angiogenesis. We sought to identify which tissues are responsive to the inflammatory effect of VEGF, to investigate the mechanisms by which VEGF increases vascular permeability, and to establish whether mediators of inflammation such as platelet activating factor (PAF) play a role in this regard. Intravenous injection of VEGF (0.001-0.1 nmol/kg) into rats induced dose-dependent protein extravasation in vascular beds of certain tissues up to 177% over control levels. In some tissues, the elevation in vascular permeability in vivo was abolished completely by selective PAF-receptor antagonists. Moreover, VEGF (10(-11) to 10(-9) M) was found to increase PAF synthesis in cultured bovine aortic endothelial cells up to 20-fold. Our results suggest that VEGF increases vascular permeability by inducing PAF synthesis. PMID- 9227553 TI - Regulation of hepatic blood flow during resuscitation from hemorrhagic shock: role of NO and endothelins. AB - We determined the role of nitric oxide (NO) and endothelins (ETs) in the regulation of hepatic blood flow during resuscitation from hemorrhagic shock (HS) in anesthetized rats. Volume resuscitation restored systemic hemodynamics and increased hepatic arterial and portal venous flow above baseline in the vehicle group. Presence of N omega-nitro-L-arginine methyl ester (L-NAME, 1 mg/kg) during resuscitation increased systemic vascular resistance (SVR) above baseline, prevented the restoration of hepatic arterial flow, and abolished portal hyperemia. Although the ETA+B-receptor antagonist bosentan (10 mg/kg) did not alter the systemic hemodynamic response, it abolished the hepatic arterial and portal hyperemia. The ETA-receptor antagonist BQ-610 (150 micrograms/kg) reduced SVR below baseline, allowed hepatic arterial hyperemia to occur, and further enhanced the portal venous hyperemia. This indicates that 1) NO reduces SVR and acts to preserve hepatic blood flow during resuscitation from HS; 2) ETA-receptor mediated vasoconstriction counteracts the systemic and portal hemodynamic effects of NO; and 3) simultaneous ETB-receptor stimulation enhances blood flow to the liver and may serve to modulate the ETA-receptor-mediated vasoconstrictive effects of ETs. PMID- 9227555 TI - Capillary red blood cell flow and activation of white blood cells in chronic muscle ischemia in the rat. AB - Increased activity of ischemic skeletal muscles in which functional hyperemia is impaired has been linked with capillary endothelial swelling postcapillary white blood cell (WBC) adherence. The perfusion pattern of capillaries under these conditions and time course of WBC activation is not known. Capillary microcirculation was studied by videomicroscopy at rest and after muscle contractions (1 Hz, 10 min) in extensor digi-torum longus muscles of pentobarbital sodium-anesthetized rat during the early stages of chronic ischemia (unilateral ligation of the common iliac artery for 3 days) and in ischemic muscles subjected to increased activity (7 days of ischemia or 3 days of ischemia plus indirect electrical stimulation via planted electrodes, 10 Hz, 7 x 10 min on 90 min off/day) to investigate how perfusion was affected. All ischemic muscles had more intermittently flowing capillaries than did unoperated control) muscles. Temporal heterogeneity of perfusion at rest, assessed by velocity, time spent stationary, and stop/start frequency of red blood cells, was similar to control values in ischemic muscles but greater in ischemic muscles subjected to additional activity. Hyperemic responses to contractions were severely blunted in all ischemic groups. The portion of morphologically nonspherical WBCs, taken to indicate activation, was 24 +/- 3% in venous blood after 3 days of ischemia vs. 14 +/- 1% in control muscles and increased further by 7 days (42 +/- 2%) when activated cells were also found in arterial blood. Thus increased muscular activity may exacerbate the adverse effects of ischemia on capillary perfusion, and WBC activation, evident before endothelial swelling is apparent, provides the potential as a circulating signal for capillary swelling in the ischemic and other muscles. PMID- 9227557 TI - Effects of ventrolateral medullary AMPA-receptor antagonism on pressor response during muscle contraction. AB - Effects of administering 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) at a concentration that preferentially blocks alpha-amino-3-hydroxy-5-methylisoxazole 4-propionic acid (AMPA) receptors into rostral ventrolateral medulla (rVLM) or caudal ventrolateral medulla (cVLM) on cardiovascular responses elicited during static muscle contraction were investigated using anesthetized rats. Two microdialysis probes were inserted bilaterally into either the rVLM or the cVLM using stereotaxic guides. A tibial nerve stimulation-evoked static muscle contraction for 30 s increased mean arterial pressure (MAP) and heart rate (HR) by 27 +/- 3 mmHg and 28 +/- 4 beats/min, respectively. Microdialysis of CNQX into the rVLM for 30 min attenuated the contraction-evoked increases in MAP and HR (10 +/- 2 mmHg and 12 +/- 2 beats/min). Developed tensions were similar during the contractions before and after microdialyzing CNQX. In contrast, administration of CNQX into the cVLM potentiated the muscle contraction-evoked cardiovascular responses (MAP, 25 +/- 4 vs. 39 +/- 6 mmHg; HR, 27 +/- 3 vs. 42 +/- 3 beats/min), with no change in developed tensions. Results demonstrate that AMPA receptors within the rVLM and the cVLM appear to play opposite modulatory roles in the central integration of cardiovascular responses elicited during static muscle contraction. PMID- 9227556 TI - Estrogen relaxation of coronary artery smooth muscle is mediated by nitric oxide and cGMP. AB - Estrogens are proposed to exert protection against cardiovascular disease, and evidence now suggests that this protection involves a direct vasodilatory effect. We have shown previously that estrogen relaxes endothelium-denuded porcine coronary arteries by opening the large-conductance calcium- and voltage-activated potassium (BKCa) channel of myocytes through guanosine 3',5'-cyclic monophosphate (cGMP)-dependent phosphorylation (35). The present study confirms these results and now demonstrates that this mechanism involves production of nitric oxide (NO). S-nitroso-N-acetylpenicillamine (SNAP), an NO donor, or 8-bromo-cGMP mimicked the effect of estrogen on BKCa channels. Furthermore, inhibition of NO synthase (NOS) attenuated estrogen- or tamoxifen-induced BKCa-channel activity, and this effect was disinhibited by L-arginine. Inhibition of guanylyl cyclase activity blocked the stimulatory effect of estrogen, SNAP, or L-arginine on BKCa channels. Furthermore, 17 beta-estradiol stimulated accumulation of nitrite and cGMP in coronary myocytes. Therefore, we propose that the vasodilatory effect of estrogen on the coronary circulation is mediated by NO. A portion of the beneficial cardiovascular effects of estrogen may be attributed to relaxation of vascular smooth muscle by a process that involves NO- and cGMP-dependent stimulation of BKCa channels. PMID- 9227558 TI - Variation in effects of Cs+, UL-FS-49, and ZD-7288 within sinoatrial node. AB - The effects of Cs+, UL-FS-49, and ZD-7288 on action potentials recorded from central, transitional, and peripheral sites in the rabbit sinoatrial (SA) node have been investigated. In isolated right atrial preparations, including the whole SA node, Cs+ (2 mM), UL-FS-49 (1 microM), and ZD-7288 (3 microM), decreased the spontaneous rate by 12 +/- 2% (n = 21), 16 +/- 3% (n = 10), and 13 +/- 3% (n = 10). They decreased the slope of the pacemaker potential without affecting action potential characteristics. The decrease of pacemaker slope was maximal (69 120%) in the periphery and minimal (22-25%) in the center. In experiments on small ball-like tissue specimens (approximately 0.3 mm diam), a decrease in spontaneous rate and pacemaker slope by Cs+ (2 mM) was largest (approximately 19 and approximately 47%) in tissue from the periphery and least (approximately 7 and approximately 17%) in tissue from the center. Because the hyperpolarization activated inward current (If) is reduced by each of these agents, the results suggest that this current might play a relatively minor role in the center but a more important role in the periphery of the SA node. PMID- 9227559 TI - Regional differences in the role of the Ca2+ and Na+ currents in pacemaker activity in the sinoatrial node. AB - The effect of block of the L-type Ca2+ current by 2 microM nifedipine and of the Na+ current by 20 microM tetrodotoxin on the center (normally the leading pacemaker site) and periphery (latent pacemaker tissue) of the rabbit sinoatrial node was investigated. Spontaneous action potentials were recorded with microelectrodes from either an isolated right atrium containing the whole node or small balls of tissue (approximately 0.3-0.4 mm in diameter) from different regions of the node. Nifedipine abolished the action potential in the center, but not usually in the periphery, in both the intact sinoatrial node and the small balls. Tetrodotoxin had no effect, on electrical activity in small balls from the center, but it decreased the takeoff potential and upstroke velocity and slowed the spontaneous activity (by 49 +/- 10%; n = 11) in small balls from the periphery. It is concluded that whereas the L-type Ca2- current plays an obligatory role in pacemaking in the center, the Na+ current plays a major role in pacemaking in the periphery. PMID- 9227560 TI - Nitric oxide inhibits neutrophil adhesion to cytokine-activated cardiac myocytes. AB - Neutrophils play important roles in myocardial injury in which an inflammatory reaction ensues. We investigated the role of nitric oxide (NO) in neutrophil adhesion to cardiac myocytes. Isolated adult rat myocytes were coincubated with human neutrophils, and the number of neutrophils adherent to myocytes was microscopically counted. The adhesion increased up to 2.4-fold when both myocytes and neutrophils were activated by interleukin-1 beta (IL-1 beta) and platelet activating factor (PAF), respectively. The NO donors, S-nitroso-N acetylpenicillamine (SNAP) and sodium nitroprusside (SNP), inhibited the adhesion by 50% when administered to neutrophils before activation. However, when activated neutrophils were added to myocytes that had been incubated with NO donors during IL-1 beta stimulation, the inhibition of adhesion was not observed. Pretreatment of neutrophils with SNAP or 8-bromoguanosine 3',5'-cyclic monophosphate did not reduce PAF-induced CD11b/CD18 expression determined by flow cytometry, nor did simultaneous treatment of myocytes with IL-1 beta and SNAP decrease IL-1 beta-induced intercellular adhesion molecule-1 expression determined by immunofluorescence staining and enzyme-linked immunosorbent assay. Thus NO inhibits neutrophil-myocyte adhesion, mainly acting on neutrophils without quantitatively affecting the upregulation of CD11b/CD18. PMID- 9227561 TI - Mechanism of atrioventricular nodal facilitation in the rabbit heart: role of the distal AV node. AB - We investigated whether atrioventricular (AV) nodal facilitation is the result of distal AV nodal action potential shortening. Atrial and bundle of His (H) electrograms and microelectrode recordings from proximal and distal AV nodal cells were analyzed in eight superfused rabbit AV node preparations in response to two pacing protocols. In the facilitation protocol, an atrial extrastimulus (A3) was preceded by an atrial impulse (A2) introduced 300, 200, 150, or 125 ms after 30 basic beats (A1). The preexcitation protocol differed from the facilitation protocol by the addition of a premature His depolarization (h2) such that the H1-h2 interval was shorter than the H1-H2 interval. Conduction curves (A3-H3 vs. H2-A3, h2-A3, and A2-A3 intervals) were constructed. Facilitation was demonstrated in all preparations when H2-A3 was used (P = 0.02) but not in the A2 A3 format. Compared with facilitation at the same A1-A2 intervals, preexcitation, despite shortening the distal cellular action potential duration, resulted in longer A3-H3 delays (P = 0.002), shorter A2-A3 intervals, and depression of the proximal nodal cellular response. Thus facilitation does not result from altered distal AV nodal characteristics and instead is a manifestation of an uncontrolled pacing protocol-dependent modulation of proximal AV nodal function. PMID- 9227562 TI - Distribution of pressure gradients along hepatic vasculature. AB - To test the hypothesis that the hepatic sinusoidal pressure is virtually identical to the portal venous pressure (Ppv) or the abdominal vena caval pressure (Pave), microvascular pressures were measured in liver vascular networks supplied by a single portal venule near the liver surface of alpha-chloralose urethan-anesthetized rabbits. With the use of a servo-null pressure-measuring system, the pressures in terminal portal venules, sinusoids, and initial hepatic venules averaged 5.7 +/- 0.8 (SD), 5.4 +/- 0.7, and 4.7 +/- 0.6 mmHg, respectively, relative to the middle of the right atrium. The mean Ppv was 7.7 +/ 1.1 mmHg and the Pave was 3.9 +/- 0.7 mmHg. The sinusoidal pressure (Psinu) averaged 2.33 +/- 1.04 mmHg lower than the Ppv, and 1.54 +/- 0.63 mmHg higher than the Pave. The pressure gradient across the sinusoids averaged < 1 mmHg. The fractional pressure gradient from the sinusoids to the vena cava averaged 41 +/- 15% of the total Ppv to Pave gradient. We conclude that most of the transhepatic resistance cannot be attributed to a specific vascular segment or vessel type and that sinusoidal pressures are not closely similar to either the Ppv or the Pave, as is often assumed. PMID- 9227563 TI - Stage-dependent changes in membrane currents in rats with monocrotaline-induced right ventricular hypertrophy. AB - Sequential changes in action potential configuration, 4-amino-pyridine-sensitive transient outward current (Ito), and L-type calcium current (ICa) in association with hypertrophy were investigated in ventricular myocytes from rats with monocrotaline (MCT)-induced pulmonary hypertension. The tissue weight ratio of right ventricle (RV) to left ventricle plus septum 14 and 28 days after a subcutaneous injection of MCT increased by 29.7 and 77.2%, respectively. Action potential duration (APD) of RV cells from MCT rats increased progressively, prolonged by 73.2 and 92.2% on days 14 and 28, respectively. The current density of Ito in RV cells from MCT rats on day 14 (32.5 +/- 4.5 pA/pF, n = 13) was significantly larger than in controls (26.8 +/- 4.5 pA/pF, n = 8; P < 0.05). On day 28, however, Ito density in MCT rats (15.3 +/- 4.6 pA/pF, n = 9) was significantly less than in controls (27.3 +/- 4.2 pA/pF, n = 10; P < 0.05). There were no differences in the voltage dependence of steady-state activation and inactivation of Ito between MCT and control rats. ICa density in MCT rats on day 14 (15.7 +/- 2.6 pA/pF, n = 10) was significantly larger than in controls (10.0 +/- 2.3 pA/pF, n = 10; P < 0.05), but there was no significant difference in Ito density between MCT rats (8.3 +/- 3.7 pA/pF, n = 10) and controls (11.6 +/- 3.0 pA/pF, n = 10) on day 28. These findings suggest that hypertrophy of mammalian hearts may cause stage-dependent changes in Ito and ICa density of ventricular myocytes. The APD prolongation in the early stage of hypertrophy may be caused mainly by an increase in ICa density, whereas the APD prolongation in the late stage may be ascribed to a reduction in Ito density. PMID- 9227564 TI - Interactions between sympathetic and vagal cardiac afferents in nucleus tractus solitarii. AB - Epicardial application of hydrogen peroxide (H2O2) reflexly elicits a sympathetically mediated pressor response. This pressor response is augmented by vagotomy and abolished by sympathectomy, suggesting an occlusive interaction between the afferents in the central nervous system (CNS). To support this observation we recorded sympathetic efferent responses from the sympathetic chain (T1-T2) before and after epicardial application of H2O2 in six cats before and after vagotomy. Cardiac sympathetic efferent responses to H2O2 were increased by vagotomy. Thus there exists an occlusive interaction between the two afferent pathways in the CNS. Because cardiopulmonary vagal afferents make their first central synapse in the nucleus tractus solitarii (NTS), we further hypothesized that cells in the NTS receive convergent inputs from sympathetic and vagal afferents and that the inputs would interact in an occlusive manner. In alpha chloralose-anesthetized sinoaortic-denervated cats, cardiac sympathetic and vagal branches were stimulated electrically at 1 Hz, either separately or in combination. Extracellular single-unit activity was recorded in the NTS. Vagal stimuli most frequently (38%) diminished sympathetically evoked unit activity ( 46.6 +/- 6.0%) versus control (1.4 +/- 1.5%). However, a few (21%) vagal and sympathetic afferent inputs were found to be additive or facilitative. We conclude that interactions occur between cardiac sympathetic and vagal afferents in the NTS. It is possible that this occlusive interaction explains the alteration in cardiac sympathetic outflow after epicardial stimulation with H2O2. PMID- 9227565 TI - Apoptosis in circulating PMN: increased susceptibility in L-selectin-deficient PMN. AB - Previous work from our laboratory has shown that polymorphonuclear leukocytes (PMN) lose L-selectin as they age in the circulation. The present study was designed to examine the relationship between PMN age and susceptibility to apoptosis in the circulation using L-selectin as a marker of PMN age in rabbits. L-selectin-deficient leukocytes were separated from a mixed population of PMN in leukocyte-rich plasma using magnetic beads. Apoptosis was measured both with both morphological criteria and by determining the level of DNA fragmentation. The L selectin-deficient cells separated in vitro showed morphological features of apoptosis (P < 0.01) and had higher levels of DNA fragmentation (P < 0.01) than the mixed population of PMN from which they were obtained. To determine if aging had a similar effect in vivo, PMN were labeled in the bone marrow with 5'-bromo 2'-deoxyuridine (BrdU) and L-selectin levels (immunocytochemistry) and DNA fragmentation (sandwich enzyme-linked immunosorbent assay) were measured in BrdU labeled PMN in peripheral blood. The results showed that the peak release of BrdU labeled PMN from the bone marrow into peripheral blood was associated with high levels of L-selectin expression, and these PMN had the lowest levels of DNA fragmentation. These results confirm that the level of L-selectin expression can be used as a marker of cell age and extend this observation by showing that aging in the circulation is associated with an increased susceptibility to apoptosis. PMID- 9227566 TI - Uncoupling of local cerebral glucose metabolism and blood flow after acute fluid percussion injury in rats. AB - We assessed local cerebral glucose metabolism (lCMRGlc) and blood flow (lCBF) interrelationships in the first hour after parasagittal fluid-percussion head injury (FPI) in rats. Matched series were studied autoradiographically for lCMRGlc and lCBF with 2-[14C]deoxyglucose and 14C-labeled iodoantipyrine, respectively. Three-dimensional autoradiographic-image mapping was to generate average data sets from which a mean ICMRGlc-to-lCBF ratio data set was derived. lCBF in neocortical regions ipsilateral to the trauma were depressed, on average, by 44% compared with sham-FPI rats, whereas contralateral lCBF values were not altered. By contrast, ICMRGlc was elevated in many cortical and subcortical sites of both hemispheres; this amounted to 1.3- to 1.4-fold increases in neocortical regions in the thalamus and 1.6- to 1.7-fold increases in the hippocampus. The lCMRGlc-to-lCBF ratio data revealed striking elevations both ipsilateral (P = 7 x 10(-7) and contralateral to the FPI (P = 0.003). The extent of metabolism-flow uncoupling, on average, amounted to 2.5-fold in the ipsilateral hippocampus and neocortex and 1.7-fold contralaterally. The loci of pronounced metabolism-flow dissociation corresponded closely to the previously documented histological distribution of neuronal necrosis. Our findings resemble events occurring in the acute focal ischemic penumbra and suggest that similar injury mechanisms may be operative. PMID- 9227567 TI - Left-to-right systolic and diastolic ventricular interactions are dependent on right ventricular volume. AB - Three-compartment elastance modeling predicts that the magnitude of gain is solely dependent on the ratio of free wall and septal elastances. However, when nonlinearities in pressure-volume relationships are considered, the same model predicts that gain is load dependent. We therefore studied left-to-right ventricular interactions in the isolated cross-perfused canine heart preparation to determine whether, in fact, right ventricular volume modulates left-to-right ventricular interaction. We found that left-to-right systolic gain increased from 0.035 +/- 0.022 to 0.073 +/- 0.017 (P = 0.003) and left-to-right diastolic gain increased from 0.067 +/- 0.050 to 0.186 +/- 0.097 (P = 0.03) in response to increased right ventricular volume. This degree of volume dependency of gain is predicted by the three-compartment model when measured nonlinearities in time varying elastance are taken into account. Future studies will need to account for changes in loading conditions when interpreting changes in systolic and diastolic interactions. PMID- 9227568 TI - Prolongation of bleeding time by acute hemolysis in rats: a role for nitric oxide. AB - The present study was aimed at clarifying the interaction between red blood cell trauma and bleeding observed in some clinical conditions. Acute hemolysis provoked by distilled water injection was followed by a significant prolongation of the "template" bleeding time in rats. Comparable effects were observed after injection of an isotonic lysate of washed red blood cells. N omega-nitro-L arginine methyl ester (L-NAME), an inhibitor of nitric oxide (NO) formation from L-arginine, normalized bleeding time when given to rats before hemolysis induction. The occurrence of hemolysis decreased ex vivo platelet adhesion to collagen without affecting platelet aggregation and induced a transient drop in blood pressure, the latter occurring during the first minute after injection. L NAME pretreatment increased ex vivo platelet adhesion but did not affect either platelet aggregation or fall in blood pressure. All the effects of L-NAME were blunted by treating the animals with the NO precursor L-arginine but not D arginine. Incubation of the erythrocyte lysate with apyrase prevented the prolongation of bleeding time induced by the hemolysate. Moreover, ADP administration, at doses that did not increase hemoglobin levels, induced effects similar to those observed after hemolysis (on template bleeding time and ex vivo platelet adhesion), which were also reversed by L-NAME and restored by L arginine. ADP is abundantly released from (hemo)lysed red blood cells and is known to stimulate release of NO, a potent vasodilator and inhibitor of platelet adhesion. ADP-dependent NO release could be responsible for bleeding time prolongation, due to abnormalities in platelet-vessel wall interaction, during acute hemolysis. Lysis of white blood cells may also contribute to prolongation of bleeding time. Because ADP could not be detected in these cells, we postulate that other mechanisms also can be involved in bleeding time prolongation after blood cell activation in vivo. PMID- 9227569 TI - Increased cardiac ppENK mRNA in cardiac hypertrophy and effects on blood pressure of its peptide products. AB - The objective of this study was to examine the expression of preproenkephalin (ppENK) in the heart in cardiac hypertrophy and the effects on cardiac contractility and blood pressure regulation of its peptide products. The ppENK derived peptides Leu5-enkephalin (LE), Met5-enkephalin (ME), Met5-enkephalin-Arg6 Gly7-Leu8 (MEAGL), and Met5-enkephalin-Arg6-Phe7 (MEAP) were administered intravenously to unanesthetized Sprague-Dawley rats and to an isolated heart preparation from the same species. LE, ME, MEAGL, or MEAP (360 nmol iv) produced an immediate decrease in heart rate, reaching its maximum within 10 s and returning to baseline by 30 s. The blood pressure response for each enkephalin was a small initial decrease followed by a marked and significant increase (P < 0.05 for MEAP). In the isolated heart preparation, neither LE, ME, MEAGL, nor MEAP altered left ventricular contractility. Cardiac hypertrophy was produced in the Dahl salt-dependent model of hypertension with a significantly greater heart weight-to-body weight ratio in the Dahl salt-sensitive (S) compared with the Dahl salt-resistant (R) rat on a high-salt diet (P < 0.05). Tissue RNA was extracted, and Northern blot analysis identified and quantitated mRNA with a 0.93-kilobase cDNA of ppENK A. There was more ppENK mRNA in the left than in the right ventricle and much less in the atria than in the ventricles. The amount of ppENK mRNA was markedly and significantly increased in the left ventricle of the Dahl S compared with the Dahl R rat (P < 0.05). In contrast, there were no differences in ppENK mRNA levels in different brain regions between the R and S rats on a high-salt diet. Interestingly, a larger ppENK mRNA of 1.75 kilobases was abundantly expressed in testicular tissue. These data showing increased ppENK expression raise the possibilities of 1) an autocrine/paracrine role for enkephalins in cardiac hypertrophy and 2) an endocrine role for the hypertrophic heart, with an increased production of enkephalins, especially MEAP, that produces vasoconstriction and further increases in blood pressure. PMID- 9227570 TI - Components of methacholine-initiated conducted vasodilation are unaffected by arteriolar pressure. AB - Conducted vasodilation occurs remotely from a site of microapplication of a drug. Intravascular pressure is required for a conducted response in vivo, yet in vitro studies in unpressurized arterioles show pressure is not essential. To determine how pressure affects conducted vasodilation, intra-arteriolar pressure was controlled within an in situ isolated segment (average length 950 +/- 96 microns, average baseline diameter 28 +/- 2.1 microns) of arterioles in the hamster cheek pouch. Methacholine (10(-4) M, 5 s) was microapplied either onto the isolated segment or remotely, with local and conducted vasodilation measured at both locations. Increasing pressure in the lumen of the segment (0-80 cmH2O) increased the segment local dilation to methacholine, and the segment-conducted dilation plateaued (at 4.1 +/- 0.8 micron) when segment pressure reached 20 cmH2O. Any local (16 +/- 1.5 microns) and conducted (4.4 +/- 1.3 microns) dilations viewed outside the segment were unaffected by segment pressure and persisted in its absence. Thus segment pressure affected only electromechanical transduction of the conducted response. Thus vasomotor signals move throughout the vasculature regardless of tone, but tone is essential to transduce the response. PMID- 9227571 TI - Regulation of adult cardiocyte growth: effects of active and passive mechanical loading. AB - Fluctuations in hemodynamic load have been documented to modulate contractile protein turnover and myofibrillar structure in the heart; however, the relative importance of active and passive loading in regulating adult cardiocyte growth remains unresolved. To address this issue at the cellular level, adult feline cardiocytes were cultured either on Silastic membranes or plastic surfaces. Cardiocyte-laden membranes were stretched 10% of their rest length to enhance passive loading, whereas heart cells cultured on plastic or Silastic were field stimulated at 1 Hz to mimic active loading. Turnover of contractile proteins and structural integrity of the contractile-cytoskeletal apparatus were monitored for periods ranging from 4 to 72 h. Active and passive loading elevated contractile protein synthesis nearly equally (approximately 50%) and promoted the attachment of remodeled myofibrils to vinculin-positive focal contacts and/or costameres during the first 24 h of loading. Thereafter, rates of contractile protein synthesis returned to control values in passively stretched heart cells but remained elevated in field-stimulated cultures. The fractional rate of growth was increased significantly (approximately 8%/day) in electrically paced cells, whereas in passively stretched cardiocytes the growth rate rose only modestly (approximately 2%/day). Changes in the rate of myocyte growth appeared more closely correlated with the development of focal contacts and myofibril remodeling than with changes in myofibrillar protein turnover per se. 2,3 Butanedione monoxime, nifedipine, and, to a lesser extent, ryanodine blocked field-stimulated contractile protein synthesis and myofibrillar remodeling but had no impact on protein turnover or myofibril reassembly in passively loaded cardiocytes. The results of these experiments imply that both active and passive loading stimulate contractile protein turnover and myofibril remodeling, but the generation of active tension accelerates cardiocyte growth to a greater extent than passive loading. Furthermore, pharmacological interventions suggest that unique pathways may mediate these cellular events in actively and passively loaded adult cardiocytes. PMID- 9227572 TI - An autocrine system for C-type natriuretic peptide within rat carotid neointima during arterial repair. AB - C-type natriuretic peptides (CNPs) are produced by endothelium and inhibit vascular smooth muscle cell (VSMC) proliferation. However, endothelial damage stimulates only transient VSMC proliferation in arteries. Here we report that a new source of CNP develops in rat carotid neointima 14 days after balloon angioplasty, when VSMC replication is subsiding despite continued absence of endothelium. CNP was detected immunohistochemically in neointimal but not medial VSMC. No other natriuretic peptides were detected immunohistochemically. CNP immunoreactivity (0.036 +/- 0.010 fmol/mg wet wt) was found in damaged arteries by radioimmunoassay, but none was detected in normal media. Reverse-phase chromatography suggested that this immunoreactivity consisted of CNP(1-53) and perhaps CNP(1-22). CNP transcript was identified by reverse transcription and polymerase chain reaction in carotid segments that had been stripped of endothelium but only once neointima had formed. Moreover, neointima expressed the NPR-C type of natriuretic peptide receptor at the same time as it synthesized CNP. Thus neointima develops an autocrine system for CNP that could regulate neointimal growth. Furthermore, the findings establish novel phenotypic differences between medial and neointimal VSMC in vivo. PMID- 9227573 TI - Open channel block of human heart hKv1.5 by the beta-subunit hKv beta 1.2. AB - Voltage-gated K+ currents in human heart are likely to derive from multisubunit complexes of pore-forming alpha-subunits with one or more auxiliary beta subunits. We recently cloned a novel beta-subunit from human atrium, hKv beta 1.2 (K. Majumder, M. De Biasi, Z. Wang, and B. A. Wible. FEBS Lett. 361: 13-16, 1995), and showed that it interacts with channels in the Kv1 family. Here we characterize the interaction of hKv beta 1.2 with hKv1.5 in terms of a two-closed state and one-open-state open channel block model. After coexpression in Xenopus oocytes, hKv1.5 currents were reduced in the presence of hKv beta 1.2, and at positive potentials an inactivation process was introduced. Deactivation kinetics of hKv1.5 were slowed, and there was an increased steepness with a -14-mV hyperpolarizing shift in the midpoint of steady-state activation. The model was able to predict all the above features of the interaction of hKv1.5 and hKv beta 1.2 as a result of rapid open channel block of activated channels. Understanding the mechanism of hKv beta 1.2 action on heart K+ channels will further aid the development of the functional and pharmacological characterization of native cardiac K+ currents. PMID- 9227574 TI - A dynamic model of ventricular interaction and pericardial influence. AB - A mathematical model describing the dynamic interaction between the left and the right ventricle over the complete cardiac cycle is presented. The pericardium bound left and right ventricles are represented as two coupled chambers consisting of the left and right free walls and the interventricular septum. Time varying pressure-volume relationships characterize the component compliances, and the interaction of these components produces the globally observed ventricular pump properties (total chamber pressure and volume). The model 1) permits the simulation of passive (diastolic) and active (systolic) ventricular interaction, 2) provides temporal profiles of hemodynamic variables (e.g., ventricular pressures, volumes, and flow) that agree well with reported observations, and 3) can be used to examine the effect of the pericardium on ventricular interaction and ventricular mechanics. It can be reduced to equivalency with models previously reported by invoking simplifying assumptions. Furthermore, model generated "dynamic interaction gains" are employed to quantify the mode and degree of ventricular interaction. The model also yields qualitative predictions of septal and free wall displacements similar to those detected experimentally via M-mode echocardiography. Such analogies may be extended easily to the study of pathophysiological states via appropriate modifications to 1) the pressure volume characteristics of the component walls (and/or pericardium) and/or 2) the specific time course of activation of the ventricular free wall or the septum. A limited number of examples are included to demonstrate the utility of the model, which may be used as an adjunct to new experimental investigations into ventricular interaction. PMID- 9227575 TI - Effect of ventricular contraction, pressure, and wall stretch on vessels at different locations in the wall. AB - A cylindrical model of the heart was used to calculate the influence of ventricular filling and (isovolumic and isobaric) contraction on the cross sectional area and resistance of a subendocardial and subepicardial maximally dilated arteriole and venule. Contraction is defined as the difference between static diastole and static systole. Furthermore, a small piece of rectangular myocardium containing the vessel was modeled to distinguish between the individual contributions of contractility (i.e., myocardial elastic properties), ventricular pressure, and local circumferential stretch to the changes in vascular area and resistance during contraction. Calculations were performed assuming the muscle fibers ran in either an apex-to-base or a circumferential direction. The results were similar for the two directions. Assuming constant, physiological arteriolar and venular pressures of 45 and 10 mmHg, respectively, coronary blood vessels were predicted not to collapse during ventricular contraction. Moreover, vascular area reduction was found to be larger for the arteriole (approximately 50%) than for the venule (approximately 30%) during both isovolumic and isobaric contractions. Consequently, arteriolar resistance was found to increase more than venular resistance (approximately 340 and 120%, respectively). Subendocardial area reductions were found to be somewhat smaller than subepicardial area reductions for the venule (by approximately 10%) but not for the arteriole. Contractility was found to be the main contributor to the changes in vascular area and resistance in the subepicardium but to contribute by < 50% to the changes in the subendocardium. Because pressure does, but stretch does not, contribute to the area change during isovolumic contraction and the reverse is true during isobaric contraction, it was concluded that although changes in vascular area and resistance may be similar for different contractions, the causes for these changes are very different. PMID- 9227576 TI - Ischemia/reperfusion-induced microvascular dysfunction: role of oxidants and lipid mediators. AB - The objective of this study was to define the role of oxidants and lipid mediators in the leukocyte-endothelial cell adhesion and albumin leakage elicited in rat mesenteric venules by ischemia-reperfusion (I/R). Intravital fluorescence microscopy was used to monitor leukocyte adherence and emigration, platelet leukocyte aggregation, mast cell degranulation, and albumin leakage after release of a 20-min arterial occlusion. I/R elicited large increases in leukocyte endothelial cell adhesion and albumin leakage. These responses were significantly attenuated in venules treated with either superoxide dismutase, oxypurinol (an inhibitor of xanthine oxidase), lodoxamide (a mast cell stabilizer), WEB-2086 (a platelet-activating factor antagonist), or SC-41930 (a leukotriene B4-receptor antagonist) but not by U-74006F (an inhibitor of lipid peroxidation). Platelet leukocyte aggregates and mast cell degranulation induced by I/R were also attenuated by administration of either superoxide dismutase or lodoxamide. These results support the hypothesis that oxidants produced, in part, by xanthine oxidase promote the formation (by mast cells and endothelial cells) of platelet activating factor and leukotriene B4, which recruit and activate leukocytes in postischemic venules. The adherent and emigrated leukocytes then mediate the increased albumin extravasation observed in the postcapillary venules. PMID- 9227577 TI - Influence of aging on protein import into cardiac mitochondria. AB - This study was undertaken to determine whether age-related changes in the content and composition of cardiac mitochondria could be due, in part, to alterations in mitochondrial protein import. Precursor proteins malate dehydrogenase and ornithine carbamoyltransferase were synthesized by in vitro transcription and translation and were incubated with mitochondria isolated from the hearts of young (4-mo), old (22-mo), and senescent (28-mo) rats. Mitochondria from senescent animals exhibited a twofold higher import rate of both precursors into the matrix compartment compared with mitochondria from young and old animals. The expression of glucose regulated protein 75 and heat shock protein 60, two matrix chaperonins that are essential for import, was elevated in the mitochondria of both old and senescent animals before the observed changes in import. Import was equally affected in senescent and young heart mitochondria by inhibition of cardiolipin, a mitochondrial phospholipid involved in protein translocation. The results indicate that the altered mitochondrial phenotype evident in the aging myocardium cannot be accounted for by reduced rates of protein import. Furthermore, levels of cardiolipin and matrix chaperonins do not appear to be rate-limiting steps in the import process. These data suggest that the protein import step of mitochondrial assembly is subject to adaptations under pathophysiological conditions. PMID- 9227578 TI - Lipopolysaccharide induces cell shrinkage in rabbit ventricular cardiac myocytes. AB - The effects of 10 ng/ml of lipopolysaccharide (LPS) on cell volume were examined in rabbit left ventricular myocytes. The myocytes were isolated with depyrogenated digestive enzymes (< 0.7 ng/ml of LPS) to minimize tolerance. Myocyte cross-sectional area (CSA) did not change after 1 h of LPS. However after 8 h, the CSA decreased to 0.93 +/- 0.01 (SE) of the baseline CSA (time = 0) in 19 LPS-exposed myocytes compared with 1.00 +/- 0.01 in 13 control myocytes (P = 0.0015). LPS-induced cell shrinkage was completely blocked by coincubation with 1 mM N-monomethyl-L-arginine, indicating a nitric oxide-mediated mechanism. Cardiac guanosine 3',5'-cyclic monophosphate (cGMP) did not change after 1 h but increased 6 h after LPS (548 +/- 31 vs. 312 +/- 20 fmol/mg protein in control cells; P < 0.05). After 8 h, bumetanide (10 microM for 30 min), a Na+/K+/2Cl- cotransport inhibitor, decreased the CSA in 15 control myocytes to 0.92 +/- 0.02 of the baseline CSA. However, in 19 myocytes with a CSA of 0.93 +/- 0.01 of baseline after 8 h of LPS, the addition of bumetanide caused no additional cell shrinkage. We conclude that low levels of LPS increase cardiac cGMP to inhibit Na+/K+/2Cl- cotransport, causing significant cell shrinkage in cardiac myocytes. PMID- 9227580 TI - Adrenomedullin microinjection into the area postrema increases blood pressure. AB - Adrenomedullin (ADM) circulates in the blood at concentrations comparable to other vasoactive peptides with established roles in cardiovascular regulation. Intravenously administered ADM produces a clear hypotensive effect, whereas intracerebroventricular microinjections result in increases in blood pressure (BP). Recently, we demonstrated that ADM influences neurons of the area postrema (AP), a central nervous system site implicated in cardiovascular control. However, to address directly the physiological significance of the actions of ADM at the AP, an in vivo microinjection study was undertaken. ADM, at two concentrations (1 and 10 microM), in volumes of 50, 100, and 200 nl, was microinjected into the AP or NTS of 21 urethan-anesthetized male Sprague-Dawley rats. Microinjection of 10 microM ADM (100 nl) resulted in significant transient (2-5 min) increases in BP [120 s area under the curve (AUC): 684.3 +/- 268.6 mmHg/s (P < 0.05)], and heart rate (HR) [AUC: 12.5 +/- 4.5 beats/min (P < 0.05)]. The lower concentration of ADM (1 microM) had no effect on either BP (179.1 +/- 143.6 mmHg/s) or HR (0.8 +/- 2.6 beats/min). ADM was also microinjected into the immediately adjacent nucleus of the solitary tract, where it was found to be without effect on either BP or HR. This study demonstrates, for the first time, a physiological role for ADM acting at a specific brain site, the AP, to produce significant cardiovascular responses. PMID- 9227579 TI - Nitric oxide increases cutaneous and respiratory heat dissipation in conscious rabbits. AB - The influence of systemic nitric oxide (NO) donor infusion and NO synthase inhibition on major thermoregulatory mechanisms was investigated under thermoneutral conditions (24 degrees C) in the conscious rabbit. Both low (25 nmol.min-1.kg-1) and high-dose (75 nmol.min-1.kg-1) infusion of the NO donors 3 morpholinosydnonimine-hydrochloride and S-nitroso-N-acetylpenicillamine augmented respiratory heat dissipation due to raised respiratory frequency (RF) and evaporative water loss (REWL). At the higher dose of NO donor, RF and REWL increased (from 107 +/- 16 to 156 +/- 19 breaths/min and from 7.12 +/- 0.97 to 11.29 +/- 1.29 mg.min-1.kg-1; P < 0.05), and, combined with a moderate rise in cutaneous heat dissipation (ear skin temperature increased from 29.03 +/- 1.76 to 33.29 +/- 2.71 degrees C; P < 0.05), deep body temperature was slightly reduced ( 0.1 degrees C, P > 0.05) without a change in metabolic heat production. In contrast, blockade of endogenous NO synthesis induced a sustained rise in body temperature (0.2 degrees C, P < 0.05), concomitant with a reduction in both RF and REWL (from 131 +/- 11 to 94 +/- 12 breaths/min and from 10.86 +/- 1.14 to 8.70 +/- 0.88 mg.min-1.kg-1, P < 0.05), whereas metabolic heat production decreased slightly and cutaneous heat dissipation was minimally altered. The data indicate that, under thermoneutral conditions, systemically applied NO primarily influences body temperature in the conscious rabbit by modulating the rate of respiratory heat dissipation, whereas the roles of cutaneous heat dissipation and metabolic heat production are relatively minor. PMID- 9227581 TI - Modulation of tissue angiotensinogen gene expression in genetically obese hypertensive rats. AB - Wistar fatty rats (WFR) show obesity and obesity-related features, including hypertension. In this study, we examined the expression of angiotensinogen mRNA in a variety of tissues at different times in WFR and control Wistar lean rats (WLR). WFR were obese and hypertensive at 16 and 24 wk. Plasma renin activity and plasma angiotensinogen concentration showed age-dependent increases in WFR but decreases in WLR. Northern blot analysis showed no significant differences in the levels of hepatic and renal angiotensinogen mRNA between WFR and WLR, and the levels of fat and adrenal angiotensinogen mRNA were lower in WFR than in WLR. On the other hand, the levels of cardiac angiotensinogen mRNA at 16 and 24 wk and those of aortic angiotensinogen mRNA at 16 wk were significantly higher in WFR than in WLR. These results show that the expression of tissue angiotensinogen mRNA is regulated differently in WFR and WLR and indicate that the development of hypertension in WFR is accompanied at least temporally with increases in plasma angiotensinogen concentration as well as in cardiac and aortic angiotensinogen mRNA. Moreover, these results suggest the existence of obesity hypertension linked and tissue-specific regulation of angiotensinogen gene expression. PMID- 9227582 TI - Involvement of cyclooxygenase-2 in LPS-induced fever and regulation of its mRNA by LPS in the rat brain. AB - We previously showed that a febrile dose of lipopolysaccharide (LPS) in rats resulted in induction of cyclooxygenase-2 (COX-2) mRNA in brain blood vessels/leptomeninges and telencephalic neurons. To elucidate the causal link between fever and LPS-induced COX-2 mRNA, we experimentally modified one or the other of these parameters and examined their relation. 1) LPS-induced fever was suppressed by pretreatment with a COX-2-specific inhibitor. 2) Levels of COX-2 mRNA in the neurons and blood vessels 2.5 h after LPS administration were even higher in the inhibitor-pretreated rats (afebrile) than in vehicle-pretreated ones (febrile). 3) After repeated administration of LPS, rats became tolerant to LPS, in which state LPS induced neither fever nor COX-2 mRNA in blood vessels/leptomeninges. When rats had not completely established LPS tolerance, they showed various degrees of fever that were closely correlated with the level of COX-2 mRNA in blood vessels but not with that in neurons. 4) Urethan anesthesia reduced basal as well as LPS-induced COX-2 mRNA in telencephalic neurons, but the rats still responded to LPS with fever and induction of COX-2 mRNA in the blood vessels/leptomeninges. These results suggest that COX-2 induced in brain blood vessels/leptomeninges is involved in the molecular mechanism of LPS-induced fever. PMID- 9227583 TI - Relationships between gastric motility and gastric vagal afferent responses to CCK and GRP in rats differ. AB - The brain-gut peptides cholecystokinin (CCK) and the mammalian bombesin-like peptide gastrin-releasing peptide (GRP) suppress food intake. Vagotomy blocks CCK but not bombesin (BN)-induced feeding suppression, demonstrating differential vagal contributions. We examined the relationship between the ability of CCK and the active fragment of GRP, GRP-(18-27), to stimulate gastric vagal afferent activity and their ability to elicit changes in gastric motility. We also examined ligated cervical vagal segments and revealed specific 125I-CCK vagal binding without evidence of radiolabeled BN binding sites. Both close arterial and intraperitoneal CCK and GRP-(18-27) produced dose-dependent increases in activity in gastric vagal mechanoreceptive afferents. CCK dose dependently decreased gastric pressure without altering antral wall tension, whereas GRP-(18 27) dose dependently increased both gastric pressure and peak antral wall muscle tension. These results suggest that GRP-(18-27) activates gastric vagal afferents secondary to its stimulation of gastric motor effects. CCK activates this same population of vagal afferents independent of changes in gastric tension, suggesting a direct action of CCK at functional vagal CCK receptors. PMID- 9227585 TI - Regulation of fluid intake in dehydrated humans: role of oropharyngeal stimulation. AB - We examined the effect of oropharyngeal stimulation on thirst, secretion of arginine vasopressin ([AVP]p), and fluid intake in six healthy adults after dehydration (28.6 +/- 1.4 ml/kg water loss) induced by mild exercise in the heat (2 h, 38 degrees C, relative humidity < 30%). Subjects performed three identical dehydration protocols followed by 75 min of rehydration at 27 degrees C consisting of 1) ad libitum drinking (Con), 2) infusion of a similar volume of water directly into the stomach via a nasogastric tube (Inf) during the first 25 min followed by combined Inf and ad libitum drinking during the remaining 50 min of rehydration; or 3) ad libitum drinking with simultaneous extraction of ingested fluid via a nasogastric tube (Ext). Plasma osmolality (Posm), [AVP]p, fluid intake, and thirst perceptions were measured throughout. On average, for all three protocols, Posm increased 7.8 +/- 0.6 mosmol/kgH2O and plasma volume decreased 4.7 +/- 1.3%, whereas thirst ratings and [AVP]p increased 7.6 +/- 1.3 cm and 3.1 +/- 0.4 pg/ml, respectively. Reflex inhibition of [AVP]p and thirst occurred within 5 min of rehydration in Con and Ext (P < 0.05) but not during Inf, supporting the hypothesis that oropharyngeal reflexes modulate osmotically stimulated thirst and [AVP]p. However, the reduction in [AVP]p during the first 5 min of Ext (-1.1 +/- 0.3 pg/ml) was less than that seen during Con (-2.1 +/- 0.4 pg/ml), suggesting that oropharyngeal stimulation is not the only factor contributing to the rapid reduction in [AVP]p during the first 5 min of drinking. During Con, subjects ingested 20.0 +/- 2.0 ml/kg of water but only drank 15% more (31.3 +/- 7.1 ml/kg) during Ext, demonstrating a clear role of oropharyngeal metering in limiting total fluid intake in humans in the presence of a persistently high dipsogenic drive. PMID- 9227584 TI - Gestational resistance to the pulmonary vasoconstrictor effect of the TxA2 mimetic U-46619: possible mechanism. AB - Pregnancy is associated with the reduction of vascular sensitivity to vasoconstrictor compounds. We have examined whether pregnancy in rabbits induces hyposensitivity of the pulmonary vascular system to U-46619. Anesthetized, mechanically ventilated nonpregnant (NP; n = 7) and late-pregnant (P; n = 7) rabbits were studied. The intravenous injection of 0.03, 0.1, and 0.3 microgram/kg U-46619 led to a dose-dependent elevation of mean pulmonary arterial pressure (MPAP) in NP rabbits from a baseline value of 15 +/- 1 to 22 +/- 1 mgHg. There was no significant MPAP response to intravenous administration of U-46619 in P rabbits. The pulmonary arterial pressure response of isolated, ventilated, and buffer-perfused lungs of P rabbits was also blunted (P < 0.001 vs. NP). Pulmonary arterial membrane binding of [125I]BOP, another thromboxane (Tx)A2 analog, indicated 48 +/- 16 fmol receptors/mg protein in P rabbits and 193 +/- 48 fmol receptors/mg protein in NP samples (P < 0.025). Receptor affinity [1/dissociation constant (KD)] was also lower in the tissue of P rabbits (P < 0.01 vs. NP). The urinary excretion of the stable TxA2 metabolite 11-dehydro-TxB2 was lower in P than in NP rabbits (P < 0.02), which made homologous desensitization an unlikely explanation for the changes of vascular TxA2 receptors. These results show that, in late gestation, rabbit pulmonary vascular sensitivity to U-46619 is reduced simultaneously with, and as a possible consequence of, downregulation of specific receptors. PMID- 9227587 TI - Carbonic anhydrase facilitates CO2 and NH3 transport across the sarcolemma of trout white muscle. AB - An isolated, perfused tail preparation was used to study the role of carbonic anhydrase (CA) in CO2 and NH3 transport across the sarcolemma of white muscle in the rainbow trout. Tissue was perfused with either control saline or saline containing the CA inhibitors quaternary ammonium sulfanilamide (QAS) or acetazolamide (Az). Inhibition of extracellular CA by QAS reduced CO2 efflux by approximately 30% and caused a significant increase in intracellular PCO2. Inhibition of total muscle CA activity (extracellular and intracellular) by Az also caused a reduction in CO2 efflux, but selective inhibition of intracellular CA only had no effect. Inhibition of both extracellular and intracellular CA activity resulted in increases in total intracellular ammonia concentrations, intracellular NH3 partial pressure (PNH3) and an increased PNH3 gradient across the sarcolemma. This suggests that both extracellular and intracellular CA function in normal NH3 transport out of the muscle. We suggest that CA in the extracellular boundary layer facilitates CO2 transport via the catalyzed hydration of CO2, thus maintaining the PCO2 gradient across the sarcolemma. H ions produced by that reaction serve to protonate excreted NH3, which helps maintain the PNH3 gradient. Thus CO2 and NH3 excretion are linked by the action of CA. PMID- 9227586 TI - Basal and stimulated nitric oxide in control of kidney function in the aging rat. AB - To investigate the activity of nitric oxide (NO) in control of renal hemodynamics during aging, studies were conducted on conscious Sprague-Dawley rats aged 3-5 mo (young, Y) and 18-22 mo (old, O). Blood pressure (BP) and renal vascular resistance (RVR) were higher in O vs. Y in control, and acute systemic NO synthesis inhibition (NOSI) increased BP and RVR, with an enhanced renal vasoconstrictor response in O. Infusion of the NO substrate L-arginine produced similar, selective renal vasodilation in both groups. The endothelium-dependent vasodilator acetylcholine caused similar falls in BP and RVR, whereas sodium nitroprusside produced an exaggerated depressor response in O vs. Y without falls in RVR in either age group. Urinary excretion of the stable NO oxidation products (NOx) decreased with age, suggesting a decline in the overall somatic NO production. In conclusion, basal tonically produced NO has a more pronounced role in maintenance of renal perfusion in aging, whereas L-arginine- and agonist stimulated renal vasodilation is not impaired with age. NO production from some source may be reduced with aging, as indicated by falls in 24-h NOX excretion, although the similarity in pressor response and enhanced renal vasoconstrictor response to NOSI suggests that the role of NO in control of total peripheral and renal vascular resistance is maintained. PMID- 9227588 TI - Effects of photoperiod and 2-deoxy-D-glucose-induced metabolic stress on immune function in female deer mice. AB - Nontropical rodents may experience large fluctuations in both food availability and energetic demands. The energy required for thermoregulation is high during the winter when energy availability is usually low. Winter conditions can induce a state of energetic stress that elevates circulating glucocorticoid levels and compromises immune function. Exposure to short days enhances immune function; the adaptive function of short-day enhancement of immune function may be to counteract the effects of stress-induced immunocompromise. To examine the role of energy availability in immune function, female deer mice were housed in either long (16:8-h light-dark cycle) or short (8:16-h light-dark cycle) days for 8 wk and then injected with either saline or 2-deoxy-D-glucose (2-DG), a glucose analog that inhibits cellular utilization of glucose and induces energetic stress. Long-day mice injected with 2-DG exhibited elevated corticosterone levels and reduced splenocyte proliferation compared with control mice. Short days buffered the animals against glucoprivation stress. Neither corticosterone levels nor splenocyte proliferation differed between 2-DG injected and control mice housed in short days. These data are consistent with the hypothesis that short days provide a buffer against metabolic stress. PMID- 9227589 TI - Long-term dietary supplementation with L-arginine prevents age-related reduction in renal function. AB - Aging is associated with loss of nephron function and reductions in serum L arginine and excretion of nitric oxide (NO) metabolites. The present study was performed to determine if long-term dietary treatment with L-arginine, the NO synthase substrate, could prevent age-related renal injury. Studies were performed in four groups of rats, aged 12-13 mo, for 8 mo: group 1 received L arginine (2% in 2.5% corn syrup, n = 5); group 2 received sodium nitrite, an NO donor (0.1%, in 2.5% syrup, n = 7); group 3 was an untreated control group (n = 7); group 4 was treated with 2.5% corn syrup (n = 5). Urinary protein increased and urinary nitrate/nitrite decreased with age in controls, but, during L arginine treatment, urinary protein decreased and nitrate/nitrite increased. Two weeks after L-arginine was stopped, urinary protein had increased and nitrate/nitrite had decreased to the same level as in controls. L-Arginine treatment increased glomerular filtration rate (GFR) by 50% compared with untreated controls. In contrast, nitrite had no effect on GFR. Morphologically, L arginine protected against aging injury by reducing the number of sclerotic glomeruli. In summary, we found that L-arginine prevented the age-related glomerular injury and reduction in GFR. The mechanism of protection, however, may be independent of NO. PMID- 9227590 TI - Vasopressin V2 receptor mRNA expression and cAMP accumulation in aging rat kidney. AB - The ability of the kidney to regulate water balance is impaired with age, although the secretion of vasopressin is maintained in senescent animals. This suggests that the cellular response to antidiuretic hormone is reduced in aging kidney. To test this hypothesis, the relationship between the expression of the vasopressin. V2 receptor mRNA and adenosine 3',5'-cyclic monophosphate (cAMP) accumulation was investigated in the medullary thick ascending limb of Henle's loop (MTAL) of adult and aging rats. Tubular suspensions of MTAL were prepared from 10- and 30-mo-old female WAG/Rij rats. The accumulation of cAMP for maximal concentration of vasopressin was 34% larger in adult than in old animals (9.5 +/- 0.5 pmol/4 min, n = 16, and 7.1 +/- 0.6 pmol/4 min, n = 12, respectively). The concentration of vasopressin corresponding to half-maximal stimulation was similar in the two groups (0.66 +/- 0.20 and 0.52 +/- 0.09 nmol, n = 5, in adult and old animals), indicating comparable sensitivity of the renal cells with age. The age-related impaired response to vasopressin of the V2 receptor was specific for females and was not observed in males. Direct stimulation of adenylyl cyclase by forskolin induced a comparable accumulation of cAMP in adult and senescent rats. The V2 receptor mRNA level in the MTAL was constant between 10 and 30 mo whether the animals were normally hydrated or dehydrated for 2 days. These data indicate that, in MTAL, the age-related impaired cAMP accumulation by vasopressin would be linked to a change either in the translation of V2 mRNA or in posttranslational processing mechanisms or in the coupling between the V2 receptor and adenylyl cyclase. PMID- 9227592 TI - Development of REM and slow wave sleep in the rat. AB - Active sleep (AS) in the neonate has been considered to be an immature form of rapid eye movement (REM) sleep. Quiet sleep (QS) has been thought to represent an immature form of slow wave sleep (SWS). To determine the relationship between the behaviorally determined states of AS and QS and electrographically determined REM sleep and SWS, we examined sleep ontogeny in the developing rat using an experimental routine that permitted long-term recordings and minimized the effects of maternal separation. Under these conditions, a transient state that included electroencephalographic slow wave activity and phasic motor activity was eventually replaced with the mature SWS pattern. Our work suggests that neonatal QS is not an immature form of SWS and that AS is best considered as an undifferentiated behavioral state from which both SWS and REM sleep develop. PMID- 9227591 TI - Characterization of an esophagocardiovascular reflex in the rat. AB - A cardiovascular reflex evoked by esophageal distension (ECR) in urethan anesthetized male Sprague-Dawley rats was studied to 1) determine whether the relevant sensory input from the esophagus is conveyed by vagal and/or spinal afferents and 2) evaluate the effects and sites of action of antinociceptive agents. Esophageal distension evoked a rise in arterial blood pressure and heart rate that increased linearly with the log of inflation pressure (25-150 mmHg). Distension (100 mmHg for 20 s) of the lower esophagus was a more effective stimulus than distension of the upper esophagus. The ECR was attenuated by unilateral and abolished by bilateral cervical vagotomy and dose dependently inhibited by morphine (1.0-4.0 mg/kg iv) or by intrathecal (T4-T5) administration of dexmedetomidine (DX, 0.05-0.5 microgram), but not by intrathecal (T4-T5) morphine (4-16 micrograms) or intrathecal (L1-L2) or intravenous DX (0.05-0.5 microgram). The ECR was also inhibited by capsaicin and by the topical administration of DX or morphine to the solitary complex. The pressor response persisted after intravenous pancuronium, scopolamine, and methscopolamine. The ECR circuit appears to consist of vagal afferents, efferent sympathetic preganglionic pathways originating in the thoracic spinal cord, and bulbospinal neurons yet to be identified. This reflex fulfills some criteria of a nociceptive event, but this interpretation requires further investigation. PMID- 9227593 TI - Recombinant leptin exchanges between parabiosed mice but does not reach equilibrium. AB - Parabiosis experiments suggest that ob/ob mice are deficient in a circulating "lipostatic" signal but respond to such a signal when it is delivered in the cross circulation from their parabiotic partner. Identification of leptin as the mutation in ob/ob mice leads to the assumption that leptin is the lipostatic signal. The objective of these experiments was to determine the circulating half life of leptin and to demonstrate whether it exchanged between parabiosed mice. Measurement of disappearance of recombinant leptin from serum in SWRJ mice indicated a circulating half-life of approximately 36 min. Single ob/ob mice or one member of a parabiosed pair of ob/ob mice received 50 micrograms recombinant murine leptin in two intraperitoneal injections a day for 10 days, starting 40 days after parabiosis surgery. Control mice and pairs received equivalent injections of vehicle. In single mice, leptin significantly reduced food intake, body weight, serum insulin, and pancreatic and liver weight. Leptin treatment of one member of a parabiosed pair of ob/ob mice reduced serum insulin, gut content (an index of food intake), and body fat in both partners. The injected parabiont lost more fat than its partner, and body temperature was increased only in the injected mouse, indicating that leptin did not reach equilibrium in the two animals. This was confirmed by Western blot analysis of serum leptin measured 2 h after injection. Therefore, although leptin can exchange between parabionts, its half-life is inadequate to allow equilibrium when a large concentration gradient exists between partners. PMID- 9227594 TI - Loss of body fat in lean parabiotic partners of ob/ob mice. AB - The objective of this experiment was to confirm whether changes in serum leptin and leptin expression were consistent with it being the "lipostatic" factor implicated by earlier parabiosis studies. Lean (+/?) and obese (ob/ob) female C57B1/6J-ob mice were parabiosed (lean-ob/ob) at 7 wk of age. Controls were ob/ob ob/ob and lean-lean pairs, and single lean and ob/ob mice. Pairs were maintained for 50 days. In ob/ob members of lean-ob/ob pairs serum insulin was normalized, food intake was suppressed, and body fat was reduced by 14%. Lean partners of ob/ob mice had a reduced rectal temperature and experienced a 37% reduction in body fat. Despite loss of fat, serum leptin and adipose leptin mRNA expression were unchanged in lean partners of ob/ob mice. These results suggest that, in lean-ob/ob parabiotic pairs, the ob/ob mouse responds to leptin originating in the lean parabiont, whereas the lean partner responds to a circulating signal, originating in the ob/ob mouse, that maintains leptin expression at inappropriate levels for the degree of adiposity of the lean animal. PMID- 9227596 TI - Effect of a cholecystokinin-A receptor blocker on protein-induced food intake suppression in rats. AB - To test the hypothesis that only dietary protein (Pro; chicken egg albumin) and not amino acids (AA), carbohydrates (CHO; cornstarch), or fats (Fat; corn oil) produces a satiating effect via cholecystokinin (CCK) receptors, devazepide was coadministered with each macronutrient. Given alone (in 4 ml ig), Pro (0.75, 1.0, and 1.5 g), AA (1.0 g), CHO (1.4 g), and Fat (2.4 g) suppressed (P < 0.05) food intake in the first hour by 0.5, 0.8, and 1.1 g; 1.9 g; and 1.0 g, respectively, and in the first 2 h of feeding by 0.8, 1.2, and 1.3 g; 1.7 g; 0.7 g; and 1.5 g, respectively. When coadministered with devazepide (0.5 mg/kg body wt ig), the suppression of food intake caused by Pro, AA, CHO, and Fat treatments was modulated in the first hour by 60, 50, and 55%; 16%, 11%; and 10%, respectively, and during 0-2 h by 63, 92, and 54%; 29%; 0%; and -20%, respectively. Devazepide (0.5 mg/kg) given intraperitoneally also modulated Pro intake suppression during the first hour by 33% and during 0-2 h by 79%. Devazepide appears to interact with mechanisms of Pro-induced food intake suppression, but not AA-, Fat-, or CHO induced food intake suppression. These studies provide evidence that CCK receptors play a role in Pro (albumin)- but not AA-, CHO (corn starch)-, or Fat (corn oil)-induced food intake suppression in rats. PMID- 9227595 TI - Role of endogenous angiotensin II on sympathetic reflexes in conscious rabbits. AB - In the present study we sought to determine the contribution of endogenous brain stem angiotensin to renal sympathetic reflexes in conscious rabbits. Initial studies determined the subtype of receptor involved in the pressor response to angiotensin II (ANG II) administration into the fourth ventricle (4V). The AT1 antagonist losartan (0.001-10 micrograms 4V) had no effect on blood pressure alone but caused a dose-dependent blockade of the pressor effect of ANG II, with complete blockade produced by 10 micrograms, an effect that lasted for at least 3 h. The AT2 antagonist PD-123319 (0.1-1,000 micrograms) and vehicle had no effect on the ANG II pressor response. The effect of losartan (10 micrograms) on the baroreceptor, chemoreceptor, and trigeminal reflexes was examined in eight rabbits that had been implanted with 4V catheters and an electrode for recording renal sympathetic nerve activity (RSNA) 1 wk earlier. Baroreflex assessments were made during normoxia and two conditions of hypoxia (10% O2 and 10% O2 + 3% CO2) before and after 10 micrograms losartan or vehicle, on separate experimental days. During normoxia and hypoxia+CO2 losartan increased resting RSNA, the range, and upper plateau of the RSNA-MAP baroreflex curves. By contrast the marked increase in RSNA due to activation of trigeminal afferents was not affected by losartan. In conclusion the effect of losartan to increase RSNA activity in conscious rabbits, particularly during hypoxia and baroreceptor unloading, suggests that endogenous ANG II via AT1 receptors normally inhibits renal sympathetic baroreceptor and chemoreceptor reflexes. PMID- 9227597 TI - Enzyme adaptation along a heterothermic tissue: the visceral retia mirabilia of the bluefin tuna. AB - We measured enzyme activities along a heterothermic tissue, the visceral retia mirabilia of the bluefin tuna, to test current theories of enzyme temperature adaptation. The heterothermic tissue model is ideal for the study of fundamental temperature adaptation because it eliminates confounding effects of whole animal acclimation. Enzymes were measured at six positions along the rete at four temperatures (15, 20, 25, and 30 degrees C). Five enzymes (aspartate aminotransferase, citrate synthase, glucose-6-phosphate dehydrogenase, glutamate dehydrogenase, and pyruvate kinase) exhibited a significant positive compensatory effect, with activity at the cold end of the rete 1.2-3.1 times higher than at the warm end. Two enzymes (alanine aminotransferase and lactate dehydrogenase) exhibited no significant compensation. On the basis of activation energies of enzymes along the rete, differences in activity were due to differences in enzyme concentration and not isozymes or enzyme modification. Analysis of the compensatory responses of the enzymes in light of their thermal sensitivities leads us to conclude that the pentose phosphate shunt is especially enhanced at the cold end of the rete. PMID- 9227598 TI - Identification and characterization of the zeta-opioid receptor in developing rat heart. AB - Opioid growth factor (OGF; [Met5]enkephalin) inhibits DNA synthesis of epicardial and myocardial cells in the neonatal rat heart in a receptor-mediated fashion. Ligand binding assays using newborn rat heart and [3H][Met5]enkephalin were performed to characterize the receptor responsible for the cell replicative effects of OGF in developing heart. Specific and saturable binding was detected, and Scatchard analysis revealed that the data were consistent for a single binding site with a binding affinity of 6.8 +/- 0.3 nM and a binding capacity of 21.8 +/- 1.9 fmol/mg protein. Subcellular fractionation studies revealed that binding was restricted to the nuclear fraction. Competition experiments showed that cold [Met5]enkephalin was the most effective ligand at displacing [3H][Met5]enkephalin. Binding was recorded on gestation days 18 and 20, reached its highest level at birth, and steadily decreased from postnatal days 1 to 15; binding at days 21 and 35 and in adults was negligible. The function, pharmacological and biochemical characteristics, distribution, and subcellular location of this OGF receptor in developing mammalian heart are consistent with the zeta-opioid receptor. PMID- 9227599 TI - Splenectomy impairs lymphocytosis during maximal exercise. AB - To evaluate the role of the spleen for the exercise-induced lymphocytosis, six splenectomized subjects and six matched control subjects cycled for 12 min at two submaximal work rates corresponding to 50 and 75% of their maximal work capacity, followed by a supramaximal intensity maintained until exhaustion (16 +/- 1 min; mean +/- SE). Venous blood samples were taken before, during, and 2 h after the maximal load. In both groups, the concentration of lymphocytes became elevated during exercise, but the increase from the level at rest was impaired in the splenectomized subjects compared with that of the controls (118 +/- 34 vs. 238 +/ 38%; P < 0.05). This was reflected in several lymphocyte subsets: cluster designation (CD) 3+ cells (pan T lymphocytes), 69 +/- 19 vs. 204 +/- 37%; CD8+ cells (T lymphocyte subset), 164 +/- 41 vs. 467 +/- 68%; CD16+ cells [natural killer (NK) cells], 291 +/- 88 vs. 870 +/- 177%; CD56+ cells (NK cells), 301 +/- 108 vs. 753 +/- 187%. Also, the specific NK cell lysis of target cells (NK cell activity) during exercise was lower for the splenectomized subjects (30 +/- 7%) than that of the control subjects (52 +/- 10%), but evaluation of lytic units indicates that this was due to a reduced number of NK cells in the assay rather than insufficient cell lysis. Plasma catecholamines reached the same level in the splenectomized subjects and control subjects, which was taken to reflect that the activity of the sympathetic nervous system was similar in the two groups of subjects. Thus the major finding of this study is that the spleen is important for lymphocytosis during exercise, accounting for two-thirds of the increase in T lymphocytes and NK cells. PMID- 9227600 TI - Epidermal growth factor and fluid and electrolyte balance in the rat. AB - Epidermal growth factor (EGF) has been shown to induce a renal diuresis and natriuresis in sheep and stimulate prostaglandin synthesis from inner rat medullary collecting duct cells in culture. The aims of our study were 1) to investigate whether the renal effects of intravenous infusion of EGF were species specific and 2) to determine the mechanism of these effects by studying the interaction between EGF and indomethacin (a prostaglandin synthase inhibitor) in the conscious rat. Sprague-Dawley rats received intravenous infusions of either 0.9% saline or 0.2 or 2.0 micrograms EGF.kg-1.h-1 over a 6-day period after an initial baseline period. Infusion of 2.0 micrograms EGF.kg-1.h-1 caused an increase in urine volume (baseline: 5.5 +/- 0.2 ml to day 5: 9.0 +/- 0.4 ml, P < 0.01) and corresponding polydipsia, but not natriuresis. Administration of indomethacin with 2.0 micrograms EGF.kg-1.h-1 attenuated (P < 0.05) the diuretic (day 5 EGF + vehicle: 12.2 +/- 1.1 ml vs. EGF + indomethacin: 8.7 +/- 0.9 ml) and polydipsic effects of EGF. These studies demonstrate that intravenous infusion of EGF causes a diuretic effect in rats without natriuresis and that prostaglandins play a role in the diuretic effect of EGF in the rat. PMID- 9227602 TI - Convection as one of the limiting factors of human respiration during normoxic exercise. AB - Each of the pathways within respiration has been suspected of limiting maximal performance, suggesting that O2 transport may be affected by each single pathway. The use of the stable, isotopic O2 molecules 16O2 and 16O18O is presented as a novel method for assessing respiration. Because of their different molecular weights, 16O2 diffuses 3% more rapidly than 16O18O, whereas 16O2 is convectively transported as rapidly as 16O18O. This can be quantified by using the overall fractionation factor alpha O. The more diffusion becomes limiting, the more 16O2 is transported in preference to 16O18O and alpha O is increased to 1.03. By contrast, the more respiration is limited by convection, the closer alpha O comes to 1.0 during the entire O2 transport. Six untrained subjects underwent normoxic exercise on a cycle ergometer. Isotopic analysis was performed at rest and during exercise loads of 50, 100, 150, 200, and 250 W using respiratory mass spectrometry. With increasing workload, a decrease in alpha O from 1.0072 at rest to 1.0033 at 250 W was determined in all subjects. On the basis of a serial resistance model of respiration, we concluded that, in our subjects, O2 transport was increasingly affected by convection but decreasingly limited by diffusion. The relative contribution of convection to the entire resistance to O2 flow ranged from > or = 44.6% at rest to > or = 74.6% at the most strenuous level of exercise, whereas the diffusive pathways decreasingly contributed to resistance to O2 flow by < or = 24% at rest and < or = 11% at 250 W. PMID- 9227601 TI - Role of endogenous ANG II and AT1 receptors in regulating arterial baroreflex responses in newborn lambs. AB - The present study was designed to test the hypothesis that endogenous angiotensin II (ANG II) influences baroreflex control of heart rate (HR) and renal sympathetic nerve activity (RSNA) early in life and to determine whether these actions are mediated by angiotensin AT1 or AT2 receptors. To test this hypothesis, we studied the effects of systemic and central administration of losartan, a selective AT1 receptor antagonist, and PD-123319, a selective AT2 antagonist, on baroreflex-mediated control of HR and RSNA in conscious newborn lambs. Systemic administration of losartan decreased resting mean arterial blood pressure (MABP) from 70 +/- 3 to 58 +/- 4 mmHg (P < 0.05) without producing reflex increases in HR or RSNA. The baroreflex response curves were shifted to the left as indicated by a decrease in the arterial pressure at the midpoint of the curve for HR (83 +/- 3 to 75 +/- 4 mmHg) and RSNA (74 +/- 2 to 69 +/- 3 mmHg; P < 0.05 for both). Losartan also reset HR and RSNA baroreflex curves when changes in baseline blood pressure were prevented by simultaneous infusion of phenylephrine. In contrast, a sustained decrease in arterial pressure of 10-12 mmHg with nitroprusside failed to shift the baroreflex function curves. PD-123319 had no effect on baseline HR, MABP, RSNA, or baroreflex responses. Lateral ventricle administration of losartan but not PD-123319 also produced a decrease in arterial pressure (81 +/- 4 to 73 +/- 3 mmHg, P < 0.05) and reset the baroreflex for HR and RSNA toward lower pressure. These results demonstrate that, early in life, endogenous ANG II exerts a tonic effect on baroreflex control of HR and RSNA to shift the curves toward higher pressure levels. The alterations in arterial baroreflex function appear independent of direct ANG II effects on arterial pressure and are mediated by AT1 receptors. PMID- 9227604 TI - Measurement of NO synthesis in humans by L-[15N2]arginine: application to the response to vaccination. AB - Induction of nitric oxide (NO) synthesis is a key element of the inflammatory response in humans. We describe a sensitive gas isotope ratio mass spectrometric (GIRMS) method for measuring urinary [15N]nitrate production during intravenous infusion of L-[guanidino-15N2]arginine and its application to investigate the effects of a controlled inflammatory stimulus, typhoid vaccination, on NO synthesis in humans. Intravenous infusion of L-[15N2]arginine at 5-12 mumol.kg 1.h-1 for 24 h in three subjects was used to determine arginine and nitrate pool kinetics. Eight subjects received primed constant infusion of 2.5 mumol.kg-1.h-1 of L-[15N2]arginine for 12 h once before and again after typhoid vaccination. NO synthesis was calculated from 15N enrichment of plasma arginine and urinary nitrate, measured by gas chromatography mass spectrometry and GIRMS, respectively, and total urinary nitrate excretion. Baseline NO synthesis was 298 +/- 44 nmol.h-1.kg lean body mass-1, representing 0.41% of arginine flux. After vaccination, NO synthesis (267 +/- 77 nmol.h-1.kg-1) was not increased (P = 0.18), despite demonstration of an acute phase response. Typhoid vaccination is not accompanied by accelerated NO synthesis. PMID- 9227603 TI - Central lipopolysaccharide elevates plasma IL-6 concentration by an alpha adrenoreceptor-mediated mechanism. AB - High circulating levels of interleukin-6 (IL-6) are evident after intracerebroventricular injection of lipopolysaccharide (LPS). To investigate the pathway of centrally induced IL-6 production, in the present study we evaluated the effects of specific alpha-adrenergic receptor antagonists administered peripherally on IL-6 production and hypertriglyceridemia induced by LPS administered centrally. In the first study, adult male Wistar-Furth rats were injected intracerebroventricularly with LPS. Centrally injected LPS increased plasma IL-6 and triglycerides (TG) in a dose-dependent fashion. To determine if LPS increased plasma IL-6 and TG through an alpha-adrenoreceptor mechanism, rats were pretreated intraperitoneally with either vehicle, phentolamine (alpha 1- and alpha 2-receptor antagonist), prazosin (alpha 1-receptor antagonist), or yohimbine (alpha 2-receptor antagonist). Thirty minutes later, rats were injected intracerebroventricularly with LPS. Whereas prazosin and yohimbine attenuated the increases in plasma IL-6 caused by LPS, phentolamine completely blocked the peripheral effects of central LPS. These data suggest that increased sympathetic activity and subsequent activation of alpha 1- and alpha 2-adrenergic receptors are important for controlling peripheral metabolic and endocrine systems when inflammatory stimuli are present in the brain. PMID- 9227605 TI - Water deprivation and rat adrenal mRNAs for tyrosine hydroxylase and the norepinephrine transporter. AB - Experiments were performed in rats to test the hypothesis that adrenal mRNA levels of tyrosine hydroxylase (TH) and the norepinephrine transporter (NET) would be modified by water deprivation via activation of the sympathetic nervous system. TH and NET mRNA levels were measured using the ribonuclease protection assay. Adrenal TH mRNA was higher (P < 0.001) in water-deprived (921 +/- 39 fg/microgram total RNA) compared with the water-replete rats (657 +/- 45 fg/microgram total RNA). In contrast, water deprivation decreased (P < 0.01) adrenal NET mRNA levels (275 +/- 66 vs. 433 +/- 63 fg/microgram total RNA). The dehydration-induced increase in TH mRNA was prevented by prior splanchnicectomy, but the decrease in NET mRNA was produced even in the absence of adrenal nerves. Water deprivation also increased (P < 0.05) plasma adrenocorticotropic hormone (84 +/- 16 vs. 42 +/- 14 pg/ml) and corticosterone (358 +/- 87 vs. 44 +/- 15 ng/ml) levels. Interestingly, the corticosterone response was reduced (P < 0.05) by unilateral adrenal denervation. These results suggest that water deprivation increases both adrenal medullary and adrenocortical activity at least in part by stimulation of sympathetic nerve activity. PMID- 9227606 TI - Intracellular acidification inhibits opposum kidney cell phosphate uptake. AB - The aim of our study is to examine the effect of intracellular pH (pHi) on inorganic phosphate (Pi) uptake by a proximal tubular cell line, the opposum kidney (OK) cells. The OK cell pHi (7.48 +/- 0.02; n = 12) was altered to levels between 6.5 and 8.5 by the high-K+ nigericin method, and cell uptakes were measured at 7.5 extracellular pH. It was found that pHi acidification suppressed Pi uptake with a decrease in maximal reaction rate, whereas alkalinization had no significant effect. Other Na(+)-dependent transport systems for glucose and amino acid were not affected by these pHi changes. The inhibition of cell Pi uptake by pHi acidification was not prevented by protein synthesis inhibitors (actinomycin D or cycloheximide) or by Na+/H+ exchange inhibitor [5-(N-ethyl-N-isopropyl) amiloride]. pHi acidification caused a significant decrease in cellular adenosine 3',5'-cyclic monophosphate (cAMP) content, and cAMP-dependent protein kinase inhibitor (H-89) also did not prevent inhibition of cell Pi uptake by pHi acidification. However, pHi acidification stimulated protein kinase C (PKC) activity and inhibition of PKC by PKC inhibitors (bisindolylmaleimide, calphostin C, or staurosporine) or prolonged exposure to phorbol ester abrogated the inhibitory effect of pHi acidification on cell Pi uptake. In summary, these studies showed that pHi acidification inhibits Pi uptake in OK cells, probably through PKC activation. These effects of pHi acidification may thus contribute to increase acid excretion in systemic acidosis. PMID- 9227607 TI - Renal, hormonal, and cardiovascular responses to chronic angiotensin I infusion in the ovine fetus. AB - Long-term infusion of angiotensin I (ANG I) into the ovine fetus has been shown to cause excess accumulation of fetal fluid in the allantoic compartment. It was hypothesized that this resulted from sustained increases in fetal urine production, and the hormonal basis was examined. ANG I (6.7 micrograms/h, n = 6) or isotonic saline (n = 6) was infused for 3 days into chronically cannulated ovine fetuses (112-122 days of gestation). ANG I caused an immediate and progressive increase in mean arterial blood pressure (from 42 +/- 2 to 57 +/- 4 mmHg), increased urine flow rate (from 15 +/- 3 to 48 +/- 8 ml/h), and increased glomerular filtration rate (from 97 +/- 15 to 146 +/- 24 ml/h), without significant changes in fetal plasma concentrations of aldosterone, atrial natriuretic factor (ANF), adrenocorticotropin, or cortisol. There were substantial increases in sodium and chloride excretion, due to both increased fetal urine concentrations and fetal urine flow, without significant changes in urine osmolality (from 134 +/- 9 to 147 +/- 12 mosmol/kg water). There were no significant changes in any parameter in the saline-infused fetuses. Neither amniotic or allantoic fluid volume was significantly changed by ANG I infusion, but allantoic fluid Cl- concentration increased significantly. The conclusions are that ANG I caused a diuresis and natriuresis in the fetal sheep independent of changes in cortisol or ANF. PMID- 9227608 TI - Histaminergic contribution to the metabolic effects of neuroglucopenia. AB - We examined the contribution of central histamine receptor (H1 and H2) blockade to the glucoregulatory responses to intracerebroventricular 2-deoxy-D-glucose (2 DG) in conscious dogs. Intracerebroventricular 2-DG (2.5 mg.kg-1.min-1 for 15 min) increased plasma glucose (2-fold), blood lactate (4-fold), and glycerol (2 fold) levels. The rate of hepatic glucose production (Ra), determined isotopically, was increased two-fold. Significant increases over basal were also noted in plasma epinephrine, norepinephrine, insulin, glucagon, and cortisol. Pretreatment with cyproheptadine and cimetidine (100 micrograms each icv 15 min before 2-DG) attenuated the 2-DG-induced hyperglycemia by approximately 50% and delayed and attenuated the increase in glucose Ra (approximately 85% vs. 2-fold in group 1). Pretreatment with H1 and H2 antagonists inhibited the increases in epinephrine, norepinephrine, and glucagon in response to neuroglucopenia but did not affect the cortisol response. These findings suggest that some of the metabolic effects of neuroglucopenia, particularly the hyperglycemic response, the increased hepatic uptake of gluconeogenic precursors, and the enhanced glucose Ra, are partly mediated through central histaminergic receptor activation. This appears to be through effects of histaminergic activation on the autonomic and hormonal responses to central neuroglucopenia. PMID- 9227609 TI - Meal size and number: relationship to dopamine levels in the ventromedial hypothalamic nucleus. AB - Evidence shows a reciprocal relationship exists between the lateral hypothalamic area (LHA) and the ventromedial hypothalamic nucleus (VMN) in food intake regulation. Since a direct correlation between meal size and LHA-dopamine in Fischer rats was previously reported, we tested the hypothesis that an inverse association may exist between meal size and VMN-dopamine response. This was studied in awake 24-h food-deprived rats who were then allowed to eat freely for 20 min while the VMN-dopamine response was measured by microdialysis every 20 min for 2 h. In a second experiment, only one-half the amount freely eaten was provided during microdialysis. The following were observed. 1) Dopamine concentrations in VMN decreased during eating. 2) The degree and duration of decrease after the meal corresponded to the size of the meal. 3) When the decreased postmeal VMN-dopamine level had returned to baseline and food was available, rats ate once more. The findings show that, in normal rats, eating was associated with decreased dopamine levels in the VMN and was followed by a lag time during which no additional eating occurred. VMN dopamine levels thereby contribute to determining the duration of the intermeal interval and hence, by inference, the meal number. PMID- 9227610 TI - Influence of route of delivery and ambient temperature on thermoregulation in newborn lambs. AB - We examined the effect of route of delivery on brown adipose tissue (BAT) function and thermoregulation in lambs born either vaginally at term or by cesarean section close to term. Immediately after birth, lambs were placed in a warm (30 degrees C; WD) or cool (15 degrees C; CD) ambient temperature, and measurements of colonic temperature, plus heat production (i.e., oxygen consumption and carbon dioxide production), were recorded for 6 h. Over the first 30 min of life, colonic temperature remained constant in vaginally delivered lambs and was lower in the WD group. Following cesarean section delivery, colonic temperature declined rapidly, a response that was greater in the CD group. Cesarean section-delivered lambs had an increased reliance on shivering thermogenesis and restored colonic temperature after 2 h, and by 6 h these parameters were higher than in lambs born vaginally. Irrespective of delivery, temperature, plasma thyroid hormone concentrations and norepinephrine content of BAT were lower in lambs born by cesarean section compared with those born vaginally. Plasma cortisol concentrations and epinephrine content of BAT were greater in lambs born by cesarean section. The amount of uncoupling protein and level of guanosine 5'-diphosphate binding in BAT were higher in vaginally delivered than in cesarean section-delivered lambs, and for each group mean values were greater for CD than WD lambs. Cesarean section delivery results in altered thyroidal, adrenal, and sympathetic activity, which appears to have a marked influence on BAT function, thereby contributing to distinct differences in thermoregulation compared with lambs born vaginally. PMID- 9227611 TI - Central infusion of the AT1 receptor antagonist losartan inhibits thirst but not sodium appetite in cattle. AB - Experiments in cattle compared the effects of intracerebroventricular (i.c.v.) infusions of losartan and PD-123319 on water intake caused by water restriction, i.c.v. infusion of hypertonic NaCl, or i.c.v. infusion of angiotensin II (ANG II). The effects of these receptor antagonists on sodium intake caused by sodium depletion were also examined. Losartan infusion caused dose-dependent inhibition of the high water intake caused by the physiological stimulus of water restriction or by ANG II infusion but did not affect salt appetite. PD-123319 infused at equimolar or greater (in ANG II experiments) doses did not affect water intake or salt intake due to sodium depletion. The results of these i.c.v. infusion experiments confirm our earlier proposal that the physiological regulation of water intake in cattle may be mediated by ANG II acting centrally via AT1 receptors. The dose of losartan that inhibited thirst in cattle did not inhibit sodium appetite, nor did an equimolar dose of PD-123319. PMID- 9227612 TI - Mechanism of the natriuretic action of gamma-melanocyte-stimulating hormone. AB - To explore the mechanism underlying the natriuretic effect of gamma-melanocyte stimulating hormone (gamma-MSH), we infused the peptide intravenously at 200 pmol/min into anesthetized rats. gamma-MSH led to a progressive increase in urinary sodium excretion (UNaV), whereas continuous infusion of the vehicle did not affect UNaV. Plasma immunoreactive gamma-MSH was nine times greater at 120 min after the start of the infusion than in vehicle-infused rats. Plasma atrial natriuretic peptide (ANP) concentration also increased as a consequence of the gamma-MSH infusion, and a strong correlation existed between the concentrations of the two peptides (n = 17, r = 0.81, P < 0.001). Urinary excretion of guanosine 3',5'-cyclic monophosphate and adenosine 3',5'-cyclic monophosphate increased as a result of the infusion. Antiserum to rat ANP blunted the natriuresis only slightly, suggesting that the increase in plasma ANP concentration was not a critical element in gamma-MSH natriuresis. gamma-MSH had no effect on ANP release from isolated rat right atrial strips superfused in vitro. Infusion of gamma-MSH (500 fmol/min) directly into one renal artery led to an ipsilateral natriuresis without change in UNaV from the contralateral kidney. Prior denervation of the infused kidney prevented the natriuresis resulting from intrarenal infusion. Intrarenal infusion of ANP (800 fmol/min) also produced ipsilateral natriuresis, which, however, was not affected by renal denervation. These studies confirm that the natriuretic action of gamma-MSH occurs primarily by an interaction with the renal nerves. Intravenous infusion of the peptide sufficient to produce a supraphysiological plasma gamma-MSH concentration also results in an increase in plasma ANP concentration; however, this increase at best plays only a minor role in the natriuresis following intravenous gamma-MSH infusion. PMID- 9227613 TI - Ins(1,4,5)P3 receptors in cerebral arteries: changes with development and high altitude hypoxia. AB - We and others have shown that adrenergic-mediated contractile responses in cerebral vessels in vitro differ with vessel segment, with developmental age, and with high-altitude, long-term hypoxia. This is associated with significant differences in alpha 1-adrenergic receptor density and norepinephrine (NE) induced response of the second messenger inositol 1,4,5-trisphosphate [Ins(1,4,5)P3]. To test the hypothesis that vessel-specific, developmental, and hypoxic-associated contractility changes are mediated, in part, by changes in Ins(1,4,5)P3-receptor [Ins(1,4,5)P3-R] density or affinity, we performed the following study. In common carotid (Com), circle of Willis, and main branch anterior, middle, and posterior cerebral arteries (MBC) from normoxic fetal (approximately 140 days), newborn (3-5 days), and adult sheep and fetal and adult sheep acclimatized to high altitude, we quantified Ins(1,4,5)P3-R with [3H]Ins(1,4,5)P3. In normoxic Com, Ins(1,4,5)P3-R density values (fmol/mg protein) in fetus, newborn, and adult were 8 +/- 53, 150 +/- 18, and 357 +/- 21, respectively (P < 0.05). In normoxic MBC cerebral arteries, the receptor density values in the three age groups were 115 +/- 15, 105 +/- 9, 99 +/- 5 fmol/mg protein, respectively. For fetal and adult Com, high-altitude, long-term hypoxemia was associated with decreases in Ins(1,4,5)P3-R density of 32 (to 58 +/ 5) and 70% (to 109 +/- 12), respectively, from control values (P < 0.01). In MBC cerebral arteries of fetus and adult, hypoxic-associated decreases in Ins(1,4,5)P3-R density from control were 80 (to 23 +/- 3) and 47% (to 53 +/- 7), respectively (P < 0.01). Ins(1,4,5)P3 binding affinity to the receptor averaged 11.8 +/- 0.5 nM and did not vary significantly as a function of vessel type, developmental age, or hypoxia. In Com, but not in MBC, Ins(1,4,5)P3-R density increased dramatically with developmental age. This suggests that differences in Ins(1,4,5)P3-R density values may account, in part, for differences in contractile responses of the two artery types in the several age groups. In response to long-term, high-altitude hypoxia, Ins(1,4,5)P3-R density values in both fetal and adult Com and MBC decreased significantly, as did their NE-induced contraction. This suggests a cellular basis for changes in cerebrovascular contractility in response to long-term hypoxia and that Ins(1,4,5)P3-R may play a role in acclimatization responses to high altitude. PMID- 9227615 TI - Differential effects of tumor necrosis factor-alpha and -beta on rat ventromedial hypothalamic neurons in vitro. AB - The effects of tumor necrosis factor (TNF)-alpha and -beta on the spontaneous firing rate of ventromedial hypothalamic (VMH) neurons were examined in rat brain slice preparations. Of 89 neurons, 36 (40%) showed a decrease in the firing rate to -78.2 +/- 4.0% (n = 36, mean +/- SE) of the preapplication level after a bath application of 20 ng/ml (approximately 1.2 nM) of TNF-alpha. This response to TNF alpha still persisted in a low-Ca2+, high-Mg2+ medium. Six (7%) of the 89 neurons were excited and 47 (53%) were unaffected by TNF-alpha. The inhibitory responses induced by TNF-alpha were abolished in a solution that contained sodium salicylate (1.9 x 10(-8)M). In contrast, TNF-beta at a dose of 20 ng/ml (approximately 1.1 nM) increased the firing rate to +39.2 +/- 6.5% (n = 11) of the preapplication level in 11 (24.5%) of 45 VMH neurons. Two of the 45 neurons (4.5%) were inhibited and 32 (71%) were unaffected by TNF-beta. The threshold concentration of TNF-alpha to alter the VMH neuron activity was lower than that of TNF-beta. Heat-inactivated TNFs were without effect. These findings suggest that TNF-alpha and -beta act as neuromodulators in the VMH, at least partly through prostaglandin synthesis, and differentially modulate the VMH neuron activity. PMID- 9227614 TI - Dysregulated proteolytic balance as the basis of excess extracellular matrix in fibrotic disease. AB - To investigate mechanisms of tissue fibrosis, we developed a model of ovine fetal bladder fibrosis due to surgically induced obstruction. Tissues were analyzed for matrix metalloproteinases (MMPs) and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). Active MMP-2 was not detected in obstructed bladders, while latent and active forms were detected in normal bladders. MMP-1 (interstitial collagenase) activity was lower in obstructed bladders. MMP inhibitory activity was increased with obstruction, as were levels of TIMP mRNA and protein. These results indicate that the proteins responsible for collagen degradation are present in the developing bladder, and a shift in the proteolytic balance favoring inhibition of degradation occurs in a model of obstruction induced fibrosis. This altered proteolytic balance favors accumulation of extracellular matrix and decreased tissue compliance characteristic of this and perhaps other fibrotic conditions. PMID- 9227616 TI - Preterm newborn lamb renal and cardiovascular responses after fetal or maternal antenatal betamethasone. AB - The optimal dose, route of administration, and treatment-to-delivery interval necessary to induce beneficial extrapulmonary effects of glucocorticoids are not known. Pregnant ewes (127 days gestation) were randomized to receive maternal or fetal intramuscular injections of betamethasone (0.2 or 0.5 mg/kg body wt) or saline 24 h before cesarean delivery of their lambs. Three hours after delivery, low-dose maternal vs. control lamb mean arterial pressure [64 +/- 4 vs. 47 +/- 2 (SE) mmHg], glomerular filtration rate (1.7 +/- 0.2 vs. 0.7 +/- 0.1 ml.min-1.kg 1), and total renal sodium reabsorption (219 +/- 31 vs. 85 +/- 12 mueq.min-1.kg 1) were increased. Comparable increases were observed in the high-dose maternal and fetal groups without effects in the low-dose fetal group. This study provides the first quantitative data demonstrating that even short-term (24-h) antenatal betamethasone exposure alters preterm newborn cardiovascular and renal functions. These responses are route and dose dependent and are comparable to glucocorticoid induced maturational effects after longer-term antenatal exposure. PMID- 9227617 TI - Increased force development rates of fatigued mouse skeletal muscle are graded to myosin light chain phosphate content. AB - Phosphorylation of myosin regulatory light chain (R-LC) increases the Ca2+ sensitivity of cross-bridge transitions, which determine rate of force development in skinned skeletal muscle fibers. The purpose of this study was to determine whether phosphorylation of R-LC is the molecular basis for the increased force development rates (+dF/dtmax) observed in fatigued mouse extensor digitorum longus muscle (EDL) (stimulated in vitro at 25 degrees C). Parameters of twitch and tetanic force were obtained after the application of different frequency conditioning stimuli (CS), which were used to vary R-LC phosphorylation and reduce peak tetanic force (Po). Without CS, R-LC phosphorylation (in moles phosphate per mole R-LC) was not elevated above rest (0.11 +/- 0.02 vs. 0.13 +/- 0.02, respectively), and no aspect of the twitch (Pt) Po was altered. Stimulating muscles at 2.5-20 Hz increased R-LC phosphorylation in a frequency-dependent manner, from 0.23 +/- 0.04 to 0.82 +/- 0.03, respectively. Moreover, stimulation at 2.5-20 Hz potentiated Pt (range: 4 +/- 2-28 +/- 2%), increased the +dF/dtmax of potentiated twitches (range: 5 +/- 1-28 +/- 2%), and reduced Po (range: 6 +/- 1-21 +/- 1%). Higher-frequency stimulation (40 or 100 Hz) did not phosphorylate R LC or potentiate Pt or twitch +dF/dtmax further. Stimulation at 40 and 100 Hz did, however, have different effects on Po compared with 20-Hz data (Po reduced 27 +/- 2 and 11 +/- 2%, respectively). The increased +dF/dtmax of potentiated twitches observed after different CS procedures were graded to R-LC phosphorylation (r = 0.97, P < 0.001). It is concluded that phosphorylation of R LC increases extent of twitch force development in mouse EDL muscle fatigued by CS. PMID- 9227618 TI - Age-related changes in renal hemodynamics in female rats: role of multiple pregnancy and NO. AB - The objective of the present study was to evaluate 1) the effect of multiple pregnancy and aging on renal function and 2) the effect of NO inhibition on renal function in aged virgin and multiply pregnant rats. Renal hemodynamics were measured in the presence or absence of chronic (2 wk) NO synthase inhibition (nitro-L-arginine methyl ester, L-NAME) in young virgins (YV, 3-4 mo), old virgins (OV, 17-18 mo), and old retired breeders (ORB, 17-18 mo) that had sustained eight to nine pregnancies and lactations. Blood pressure was not different between control YV and OV. Glomerular filtration rate (GFR), renal plasma flow (RPF), and renal vascular resistance (RVR) were similar in OV and control YV. In contrast, the renal vasculature of ORB was more vasoconstricted in ORB than in YV or OV:GFR was decreased by 35% and RVR was higher than in YV or OV. With L-NAME there were similar increases in arterial pressure in all rats. In control YV, L-NAME had no effect on GFR, decreased RPF by 20%, and increased RVR by twofold. In OV, L-NAME decreased GFR by 30% and RPF by 60% and increased RVR by 3.3-fold. In ORB, L-NAME had no effect on GFR, decreased RPF by 30%, and increased RVR by 1.8-fold. These data suggest that the renal vasculature of ORB is vasoconstricted and that the mechanism may be due to a decrease in NO production. PMID- 9227619 TI - Differential role of glucocorticoids in mediating endotoxin-induced changes in IGF-I and IGFBP-1. AB - The purpose of the present study was to determine whether endogenous elevations in glucocorticoids mediate the changes in insulin-like growth factor (IGF) 1 and IGF binding protein (IGFBP) 1 levels in plasma and tissues observed after in vivo administration of lipopolysaccharide (LPS). In overnight-fasted male rats LPS injected via the tail vein decreased the IGF-I concentration in plasma, liver, and skeletal muscle (30-45%) and increased IGF-I content in kidney (approximately 3-fold). LPS also decreased IGF-I mRNA abundance in liver and muscle and increased gene expression in kidney. Concomitantly, IGFBP-1 levels in plasma, liver, and muscle were markedly elevated by LPS. All these changes were associated with a greater than fourfold elevation in plasma corticosterone. Pretreatment of rats with the glucocorticoid receptor antagonist RU-486 completely prevented or blunted the LPS-induced changes in IGF-I content in plasma, liver, muscle, and kidney. In liver and muscle RU-486 significantly attenuated the reduction in IGF-I mRNA abundance produced by LPS, but in kidney the LPS-induced increase in IGF-I mRNA was still evident. In contrast, pretreatment with RU-486 did not prevent or attenuate the LPS-induced increase in IGFBP-1 levels in plasma, liver, or muscle. These data suggest that glucocorticoids play a major role in regulating IGF-I mRNA and peptide content in tissues in response to LPS, but the increased IGFBP-1 in blood and tissues induced by LPS appears largely glucocorticoid independent. PMID- 9227620 TI - Cardiovascular afferent inputs to ventral tegmental area. AB - Extracellular single-unit recording experiments were done in alpha-chloralose anesthetized, paralyzed, and artificially ventilated rats to investigate the effect of selective activation of arterial baroreceptors and stimulation of cardiovascular depressor sites in the nucleus of the solitary tract (NTS) on the discharge rate of neurons in the ventral tegmental area (VTA). Electrical stimulation of the aortic depressor nerve (ADN), which is known to carry aortic baroreceptor afferent fibers only, excited 12 of 21 (mean onset latency 42.4 +/- 8.8 ms) and inhibited 2 of 21 (mean onset latency 42.5 +/- 6.5 ms) single units in the VTA. The discharge rate of VTA units was also altered during the reflex activation of arterial baroreceptors by the acute rise in arterial pressure (AP) to systemic injections of phenylephrine (10 micrograms/kg i.v.): 12 of 44 units were excited and 15 of 44 were inhibited. Units that responded to either ADN stimulation or the reflex activation of the baroreflex also responded to stimulation of depressor sites in the NTS. An additional 12 units that were found in barodenervated controls to be responsive to NTS stimulation were nonresponsive to selective activation of arterial baroreceptors. These data indicate that cardiovascular afferent inputs modulate the activity of neurons in the VTA and suggest that changes in systemic AP may exert an effect on the activity of neurons involved in mesolimbic and mesocortical function. PMID- 9227621 TI - Metabolic acid-base influences on renal thiazide receptor density. AB - The renal responses to metabolic acidosis/alkalosis involve changes in the proximal tubule, loop of Henle, and collecting ducts. We tested for acid- or base induced changes in the distal convoluted tubule (DCT) by examining the renal density of the DCT's receptor for thiazide-type diuretics (TZR), as estimated by the binding of [3H]metolazone in Wistar-Kyoto rats. TZR density significantly decreased by 17% in rats ingesting NH4Cl for 3.5 days and by nearly 30% after 7 days; TZR increased up to 40% in rats ingesting NaHCO3 for 2-4 days but was no longer significantly increased after 7 days. Urinary excretion of chloride increased as renal density of the TZR decreased, a finding consistent with the interpretation that acidosis/alkalosis not only altered TZR density but coordinately altered reabsorption of NaCl by the thiazidesensitive Na-Cl cotransporter. The result is that delivery of Na from DCT is enhanced during acidosis and decreased during alkalosis, assisting in compensatory changes in distal nephron secretion of hydrogen ion. The integrated renal response to metabolic acidosis/alkalosis involves a decrease in renal TZR with acidosis and an increase in TZR with alkalosis. PMID- 9227622 TI - Bradykinin-mediated activation of renal sensory neurons due to prostaglandin dependent release of substance P. AB - In anesthetized rats, renal pelvic administration of bradykinin results in a prostaglandin (PG)-dependent increase in afferent renal nerve activity (ARNA). We now measured renal pelvic release of PGE and substance P during renal pelvic administration of bradykinin. Bradykinin increased ARNA and renal pelvic release of PGE by 497 +/- 252 pg/min and substance P. by 10.7 +/- 7.2 pg/min. Renal pelvic perfusion with indomethacin abolished the bradykinin-mediated increase in ARNA and reduced renal pelvic release of PGE and substance P by 76 +/- 11 and 72 +/- 8%, respectively. To examine whether the increased substance P release contributed to bradykinin-mediated activation of renal sensory receptors, renal pelvis was perfused with the substance P-receptor antagonists CP-96,345, CP 99,994, or RP-67580. The ARNA response to bradykinin was reduced 73 +/- 11, 55 +/ 12, and 64 +/- 10% by CP-96,345, CP-99,994, and RP-67580, respectively. The inactive enantiomers CP-96,344 and RP-68651 had no effect. These data suggest that bradykinin increases renal pelvic release of PGE, which facilitates the release of substance P, which in turn stimulates substance P receptors. Thus the ARNA response to bradykinin is largely mediated by activation of substance P receptors. PMID- 9227623 TI - Chronic corticosterone treatment increases myocardial infarct size in rats with ischemia-reperfusion injury. AB - Experiments were conducted to determine the effect of chronic elevations in corticosterone on myocardial infarct size. Male Sprague-Dawley rats were treated for 7-22 days with corticosterone. Plasma corticosterone concentrations averaged 0.8 +/- 0.3 in control and 14.9 +/- 1.2 micrograms/dl in corticosterone-treated conscious rats. Experiments were performed in anesthetized rats. After a 30-min control period, myocardial ischemia (30 min)-reperfusion (3 h) was performed in control and corticosterone-treated rats. Mean arterial pressure (+/-SE) in control rats during control, ischemia, and reperfusion periods averaged 111 +/- 4, 100 +/- 5, and 94 +/- 4 mmHg (n = 6), respectively. Chronic treatment with corticosterone increased mean arterial pressure in all three periods 128 +/- 6, 117 +/- 7, and 109 +/- 7 mmHg; n = 8; P < 0.05). Infarct size (as % area at risk) was significantly larger in rats with chronic elevations in corticosterone compared with control rats (77 +/- 2 vs. 51 +/- 5%, P < 0.05). Acute (2 h) blockade of the glucocorticoid type II receptors with mifepristone antagonized the increases in arterial pressure and infarct size produced by chronic administration of corticosterone. Neither mifepristone nor acutely administered corticosterone affected arterial pressure or infarct size in rats without chronic corticosterone treatment. The effect of chronic elevations in plasma corticosterone concentration to increase infarct size could contribute to the increased risk of cardiovascular disease in clinical conditions associated with elevated glucocorticoid levels. PMID- 9227624 TI - Physiological responses to cold stress during prolonged intermittent low- and high-intensity walking. AB - In a previous study [Am. J. Physiol. 272 (Regulatory Integrative Comp. Physiol. 41): R226-R233, 1997], the physiological responses to 240 min of intermittent low intensity walking exercise in a cold (+5 degrees C), wet, and windy environment (Cold) may have been influenced by a 120-min preceding phase of intermittent higher-intensity exercise. Furthermore, the physiological responses observed during this latter phase may have been different if it had been more prolonged. To address these questions, active men attempted a 360-min intermittent (15 min of rest, 45 min of exercise) exercise protocol in Cold and a thermoneutral environment (+15 degrees C, Neutral) at a low (0% grade, 5 km/h; Low; n = 14) and a higher (10% grade, 6 km/h; High; n = 10) intensity. During Low, rectal temperature was lower in Cold than in Neutral, whereas O2 consumption, carbohydrate oxidation, plasma norepinephrine and epinephrine, and blood lactate were higher. During High, Cold had a similar but less marked influence on the thermoregulatory responses to exercise than during Low. In conclusion, the physiological responses to Low are similarly influenced by Cold whether or not they are preceded by High. Furthermore, during intermittent exercise up to an intensity of approximately 60% of peak O2 consumption, a cold, wet, and windy environment will influence the physiological responses to exercise and potentially impair performance. PMID- 9227625 TI - Very low frequency oscillations in arterial blood pressure after autonomic blockade in conscious dogs. AB - The aim of this study was to investigate spontaneous variability of arterial blood pressure in conscious foxhounds in the absence of direct sympathetic and parasympathetic influences. Autonomic blockade was achieved by administration of the ganglionic blocking agent hexamethonium (n = 7). In contrast to the control group (n = 7), marked oscillations with a cycle length of 100 s (0.01 Hz) were observed. The relationship of the power densities of the oscillation band (0.01 +/- 0.005 Hz) to the total power increased threefold (0.213 +/- 0.007 vs. 0.057 +/- 0.005; P < 0.01). The 0.01-Hz oscillations typically commenced after some delay. To test whether the absence of the mechanoreceptor afferents was responsible for these fluctuations, we investigated an additional group of foxhounds that were subjected to total baroreceptor and cardiopulmonary receptor denervation (n = 7). Neither in this protocol, nor in a group subjected to denervation and ganglionic blockade (n = 6), did we observe sustained oscillations in this frequency range. Since the oscillations were not seen after combined afferent (mechanoreceptor denervation) and efferent (ganglionic) blockade, central oscillators as a source of the oscillations can be ruled out. A simple model of a circulating pressoric factor may explain the fluctuations, provided that there is a time delay between the stimulus and the release or action of the factor. The findings suggest that a circulating factor accounts for the 0.01-Hz oscillations, which is dependent on intact pathways from the cardiac receptors or baroreceptors to the central nervous system. This hypothesis is put forward since cardiopulmonary and baroreceptor denervation blocked the oscillations seen after ganglionic blockade. PMID- 9227626 TI - Hemodynamic, hormonal, and renal effects of adrenomedullin in conscious sheep. AB - Adrenomedullin is a recently discovered peptide that has been shown to reduce arterial pressure and induce natriuresis. However, few studies have examined the biological actions of adrenomedullin in conscious animals in an integrative manner. Accordingly, we have examined the hemodynamic, renal, and endocrine actions of adrenomedullin infused intravenously at 10 and 100 ng.kg-1.min-1 (each 90 min) in a vehicle-controlled study in eight normal conscious sheep. Adrenomedullin reduced right atrial pressure (P < 0.05) and diastolic (15 mmHg, P < 0.01) and mean arterial pressure (10 mmHg, P < 0.05) and increased cardiac output (3 l/min, P < 0.001). Total peripheral resistance was reduced 40% (P < 0.001). Urinary sodium was reduced to 35% of control during the 90-min clearance period immediately postinfusion (P < 0.05). Adrenomedullin increased plasma adenosine 3',5'-cyclic monophosphate levels (P < 0.001). Plasma renin activity was elevated during adrenomedullin (P < 0.001) coincident with the peak hypotensive effect, whereas plasma aldosterone was not affected and plasma norepinephrine levels fell (P < 0.05). In conclusion, adrenomedullin had clear blood pressure-lowering effects with increased cardiac output and stimulation of renin but suppressed sympathetic activation in conscious sheep. The physiological implications of these findings require further study. PMID- 9227627 TI - Endotoxin impedes vasoconstriction in the spleen: role of endogenous interleukin 1 and sympathetic innervation. AB - Processes relevant for an appropriate immune response such as immune cell traffic and recirculation require a tight control of blood supply to lymphoid organs. Interactions between endogenous cytokines and sympathetic nerve fibers in lymphoid organs can contribute to this control. The results reported in this paper show that 1) administration of low doses of lipopolysaccharide (LPS), an endotoxin derived from gram-negative bacteria, causes an increase in splenic blood flow (SBF); 2) this increase is mediated by the production of endogenous interleukin-1 (IL-1); 3) the effect of LPS on SBF requires an intact splenic sympathetic innervation; 4) the LPS-induced increase in SBF is exerted at the postganglionic level; 5) the endotoxin inhibits the vasoconstriction induced by the in vivo stimulation of the splenic nerve but does not affect the vasoconstriction induced by norepinephrine (NE); and 6) although IL-1 and LPS stimulate general sympathetic activity as reflected by increased peripheral vascular resistance, they do not increase NE concentration in splenic dialysates. Together these in vivo results indicate that endogenous IL-1 affects blood supply to the spleen by inhibiting the sympathetic vasoconstrictor tonus at a postganglionic, prejunctional level. This effect is expected to be relevant for immune cell recirculation, homing, and traffic as well as antigen trapping in the spleen, an organ specialized in the control of these processes during immune responses. PMID- 9227628 TI - Microlocalization and effects of adrenomedullin in nephron segments and in mesangial cells of the rat. AB - We examined microlocalization of mRNA coding for adrenomedullin (AM), using reverse transcription-polymerase chain reaction (RT-PCR), and the effects of AM on adenosine 3',5'-cyclic monophosphate (cAMP) generation and water transport in microdissected rat nephron segments. We also examined intraglomerular site of the expression of AM and AM-stimulated cAMP generation in cultured rat mesangial cells (MC). RT-PCR demonstrated the signals for AM mRNA in glomerulus (Glm), cortical collecting duct (CCD), outer medullary collecting duct (OMCD), and inner medullary collecting duct (IMCD) but not in proximal convoluted tubule (PCT) or medullary thick ascending limb (MTAL). AM (10(-7) M) stimulated cAMP generation in Glm >> CCD = IMCD > OMCD but not in PCT or MTAL, which corresponded to the results of the expression of AM mRNA. AM (10(-8) M) slightly increased osmotic water permeability by 24% in perfused terminal IMCD. Northern blot analysis revealed high expression of AM mRNA in MC. AM (10(-7) M) stimulated cAMP generation in MC both in the presence and absence of fetal calf serum, suggesting that AM-dependent cAMP generation was evident both in cycling MC and in quiescent MC. AM may work as a diuretic peptide mainly by increasing glomerular filtration rate via cAMP in MC. PMID- 9227629 TI - Insulin-like growth factor I increases renal 1,25(OH)2D3 biosynthesis during low P diet in adult rats. AB - Dietary P restriction increases renal 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] biosynthesis through stimulation of proximal tubule 25-hydroxyvitamin D3-1 alpha hydroxylase (1-OHase). Because insulin-like growth factor I (IGF-I) is required for 1-OHase stimulation by low-P diet (LPD) and because 1-OHase stimulation by low-Ca diet and parathyroid hormone is lost with aging, studies were undertaken to determine whether 1-OHase activity during LPD is impaired with age and whether IGF-1 can increase 1-OHase activity in adult rats. Five days of LPD increased in vitro 1-OHase activity in young (97.3 +/- 13.5 vs. 49.7 +/- 6.8 pg.mg protein-1.5 min-1, P < 0.005) but not adult (42.3 +/- 5.37 vs. 41.2 +/- 8.9) rats. In LPD-fed adult rats, recombinant human IGF-I (rhIGF-I, 1.4 mg.kg body wt-1.day-1) for 72 h increased 1-OHase (65.2 +/- 5.88 vs. 95.1 +/- 7.26 pg.mg protein-1.5 min-1, P < 0.005). The results show that the rise in 1-OHase activity during LPD is lost in adult rats and that rhIGF-I can overcome the inhibition and stimulate renal 1 OHase activity to levels observed in young animals. The studies indicate that the age-related loss of 1-OHase activity is reversible. PMID- 9227630 TI - UTP and ATP induce different membrane voltage responses in rat mesangial cells. AB - UTP and ATP induce different membrane voltage responses in rat mesangial cells. Recent studies have indicated that UTP and ATP might modulate mesangial cell function in a different manner. Here we compared the effect of UTP and ATP on membrane voltage (Vm) and ion currents in mesangial cells in primary culture, and we examined whether different nucleotide receptors are involved. In patch-clamp experiments in the fast whole cell configuration, UTP (in contrast to ATP) caused a sustained and concentration-dependent depolarization (half-maximal effective dose, 10(-5) M), but ATP caused only a transient depolarization. During the depolarization, UTP induced a sustained increase of the whole cell conductance (Gm), whereas ATP induced only a transient increase of Gm. When cells were dialyzed with Cs2SO4 and extracellular Cl- was replaced by 145 mM sodium gluconate, addition of UTP or ATP (both 10(-4) M) did not significantly increase Gm. Addition of ATP in the presence of UTP caused an additional depolarization by 5 mV, which was followed by a hyperpolarization by 21 mV. Repetitive application of ATP led to an attenuation of the ATP-induced depolarization. Then, in the presence of ATP, UTP still induced a significant depolarization by 10 mV. Suramine and reactive blue 2 did not inhibit the depolarization induced by UTP, but these inhibited the Vm response to ATP. In microfluorescence experiments, UTP and ATP caused a concentration-dependent increase of the intracellular calcium activity ([Ca2+]i) in mesangial cells. Application of both UTP and ATP had no additive effect on [Ca2+]i. The results suggest that mesangial cells possess, in addition to P2y purinoceptors, separate nucleotide receptors for UTP. PMID- 9227631 TI - Molecular and functional identification of beta-adrenergic receptors in distal convoluted tubule cells. AB - Renal nerve stimulation or circulating catecholamines activate the beta adrenergic receptors that mediate direct effects on tubular transport. Three subtypes of beta-adrenergic receptors have been characterized: beta 1, beta 2, and beta 3. beta-Adrenergic-receptor effects on Na+ and Ca2+ transport in distal convoluted tubules (DCT) have not been established. The focus of this study was to 1) identify the subtypes of beta-adrenergic receptors in DCT cells and 2) examine functional responses to beta-receptor activation on adenosine 3',5' cyclic monophosphate (cAMP) formation and Na+ and Ca2+ entry. To determine the subtypes of beta-receptors present, RNA isolated from immortalized mouse DCT cells was reverse transcribed, and the cDNA was amplified using primers designed to reported sequences for beta 1-, beta 2-, and beta 3-receptor subtypes. Products of the appropriate sizes were obtained with beta 1- and beta 2-primers. No product was observed with primers to the beta 3 sequence. Receptor products were confirmed by sequencing and are identical to reported mouse beta 1- and beta 2-receptor sequence. Receptor binding of[3H]dihydroalprenolol was 123 +/- 13 fmol/mg protein, and a 3:1 ratio of beta 1- to beta 2-receptors was observed with DCT cell membranes. Isoproterenol, a beta-receptor agonist, increased cAMP formation 8.5-fold. Pretreatment with the antagonist propranolol abolished agonist-induced cAMP accumulation. Isoproterenol significantly increased 22Na+ uptake to 345 +/- 23 compared with a basal rate of 256 +/- 12 nmol.min-1.mg protein-1 and was blocked with propranolol and beta 1- and beta 2-selective antagonists. Isoproterenol had no effect on 45Ca2+ entry into DCT cells. In summary, DCT cells express three times more beta 1- than beta 2-receptors and express no detectable beta 3-adrenergic receptors. beta-Receptors couple to adenylyl cyclase, and activation of beta-adrenergic receptors increases Na+ but not Ca2+ entry in DCT cells. PMID- 9227632 TI - Hydrogen peroxide downregulates IL-1-driven mesangial iNOS activity: implications for glomerulonephritis. AB - In glomerulonephritides, autacoids such as nitric oxide (NO), reactive oxygen species, and prostanoids are produced in increased amounts in response to cytokines such as interleukin-1 (IL-1). These autacoids influence the expression of glomerular injury by their direct as well as interactive actions. We studied the effect of hydrogen peroxide (H2O2) on NO production in rat mesangial cells. We demonstrate that transient exposure of mesangial cells to H2O2 prior to sustained exposure to IL-1 decreased extracellular accumulation of NO2/NO3 and cellular guanosine 3,'5'-cyclic monophosphate (cGMP) content. H2O2 markedly impaired inducible nitric oxide synthase (iNOS) activity induced by IL-1 directly measured by the conversion of L-[14C]arginine to L-[14C]citrulline. Such impairment in iNOS activity was accompanied by a parallel reduction in iNOS protein abundance but not by a reduced expression of iNOS mRNA. The inhibitory effect of H2O2 on NOS activity was further supported by peroxide-induced impairment in IL-1-driven, NO-dependent synthesis of prostaglandin E2. Our studies thus provide the first direct evidence of a posttranscriptional inhibitory effect of H2O2 on iNOS activity. Additionally, our studies uncover the existence of a previously unrecognized effect of H2O2 on the production of NO that may exert influence on the severity of glomerular injury during glomerular inflammation. PMID- 9227633 TI - Disassociation of oxidant-induced ATP depletion and DNA damage from early cytotoxicity in LLC-PK1 cells. AB - To determine the mechanism(s) of oxidant-mediated cell lysis in renal tubular epithelial cells, we determined ATP depletion, DNA damage, lipid peroxidation, and cytotoxicity in LLC-PK1 cells exposed to 500 microM hydrogen peroxide for 1 h with and without inhibitors of lipid peroxidation including a lazaroid compound, 2-methylaminochroman (2-MAC), and Trolox, a vitamin E analog. ATP levels were determined by luciferin-luciferase, DNA damage by the alkaline unwinding technique, lipid peroxidation by the generation of malondialdehyde, and early cytotoxicity (5 h) by the release of 51Cr, whereas late cytotoxicity (24 h) was determined by release of [3H]leucine from prelabeled cells. Cells exposed to 500 microM hydrogen peroxide demonstrated significant (P < 0.01) ATP depletion, DNA damage, and lipid peroxidation, followed by cell death at 5 h. Concentrations of 0.1-25 microM 2-MAC or 25-500 microM Trolox each markedly and significantly (P < 0.01) inhibited lipid peroxidation and early cytotoxicity but had little to no effect on ATP depletion or DNA damage. Thus oxidant-stressed cells remained intact for several hours despite significant ATP depletion and DNA damage when lipid peroxidation was inhibited with the antioxidant compounds. At 24 h, 2-MAG and Trolox had lost their protective effect, suggesting that mechanisms other than lipid peroxidation play a role in later cytotoxicity. We conclude that ATP depletion and DNA damage are not the primary mediators of early cytotoxicity following oxidant stress, whereas lipid peroxidation plays an central role in mediating early cytotoxicity following oxidant injury. PMID- 9227634 TI - Potassium depletion and acid-base transporters in rat kidney: differential effect of hypophysectomy. AB - Potassium depletion is involved in the pathophysiology of metabolic alkalosis. In the present study, the expression of renal acid-base transporters that are involved in HCO3- reabsorption was studied in potassium depletion. Rats fed potassium-deficient (KD) diet developed significant hypokalemia at 14 days (serum K+ 1.9 +/- 0.2 in KD vs. 4.2 +/- 0.2 meq/l in control, P < 0.01) but not at 6 days (3.8 +/- 0.3 in KD vs. 4.1 +/- 0.3 meq/l in control, P > 0.05). Kidney mRNA for colonic H(+)-K(+)-adenosinetriphosphatase (H(+)-K(+)-ATPase, cHKA) increased by approximately 3- and 11-fold at 6 and 14 days of KD diet, respectively, indicating that increased expression preceded the onset of hypokalemia. The expression of Na+/H+ exchanger 3 (NHE-3) mRNA and its cognate protein remained unchanged at 6 and 14 days of KD diet. The mRNA levels for NHE-1, NHE-2, and NHE 4 also remained unchanged at 6 and 14 days of KD diet. Hypophysectomized (HPX) rats fed KD diet for 14 days developed similar hypokalemia. However, the expression of cHKA mRNA in the kidney was decreased by approximately 80% in potassium-depleted (HPX+KD) rats (P < 0.01 vs. KD only). Hypophysectomy did not affect the mRNA levels for either gastric H(+)-K(+)-ATPase (gHKA) or NHE isoforms in KD animals. Thus potassium depletion increases expression of cHKA in the kidney but not that of gHKA or NHE isoforms. The signal for this increase appears to precede hypokalemia. Furthermore, the data suggest that pituitary hormone(s) plays an important and novel role in the regulation of cHKA. PMID- 9227635 TI - Effect of hypokalemia on the abundance of HK alpha 1 and HK alpha 2 protein in the rat kidney. AB - An H(+)-K(+)-adenosinetriphosphatase (H(+)-K(+)-ATPase) contributes to potassium reabsorption by the collecting ducts of the rat kidney. mRNAs for two isoforms of the H(+)-K(+)-ATPase, HK alpha 1 and HK alpha 2, have been found in the rat kidney. To evaluate whether the HK alpha 1 and HK alpha 2 proteins are present in the rat kidney, microsomes enriched in HK alpha 1 or HK alpha 2 were isolated using the MiniMac magnetic separation system with antibodies directed against either HK alpha 1 (HK 12.18) or HK alpha 2 (AS 31.7). Immunoblots of rat kidney microsomal protein isolated with HK 12.18 revealed a band approximately 94 kDa in size that comigrated with the G1 fraction of the stomach. Immunoblots of rat kidney microsomal protein isolated with AS 31.7 revealed a band slightly greater than 94 kDa that comigrated with a band obtained from rat colonic microsomal protein. To examine the effect of perturbations in potassium metabolism, the abundance of the HK alpha 1 and HK alpha 2 isoforms was compared in rats fed a normal or potassium-deficient diet. A low-potassium diet increased the abundance of HK alpha 2, whereas that of HK alpha 1 was not altered. These data suggest that HK alpha 2 might be the isoform responsible for potassium conservation by the kidney. PMID- 9227636 TI - Expression of PTHrP, PTH/PTHrP receptor, and Ca(2+)-sensing receptor mRNAs along the rat nephron. AB - To provide a frame of reference for studies of renal divalent cation and phosphate metabolism, we assessed the cellular localization of kidney calcium receptor (RaKCaR), parathyroid hormone-related protein (PTHrP), and parathyroid hormone/ parathyroid hormone-related protein (PTH/PTHrP) receptor mRNA. The studies used using reverse transcription-polymerase chain reaction (RT-PCR) applied to cDNA prepared from dissected rat nephron segments and from primary cultures of mouse juxtaglomerular granular cells. With species-specific primers, PCR products of expected size were obtained for RaKCaR (967 bp), PTHrP (420 bp), and PTH/PTHrP receptor (817 bp), with product identity being confirmed by restriction digestion. RaKCaR mRNA was found in medullary and cortical thick ascending limbs (MTAL and CTAL, respectively), the macula densa-containing segment, distal convoluted tubules (DCT), and, to a lesser extent, in cortical collecting ducts (CCD). It was not found in glomeruli, proximal convoluted and straight tubules (PCT and PST, respectively), outer and inner medullary collecting ducts (OMCD and IMCD, respectively), or in juxtaglomerular granular cell isolates. PTHrP mRNA was predominantly expressed in glomeruli and at lower levels in PCT and the macula densacontaining segment but was not detectable in CTAL, MTAL, DCT, and CD segments. Presence of PTH/PTHrP receptor mRNA was demonstrated in glomeruli, PCT, PST, CTAL, MTAL, and DCT but not in CD segments. These results suggest that the function of TAL and DCT cells, in addition to being affected by PTH, may be directly altered by extracellular divalent cations through RaKCaR and that PTHrP may act in the glomerulus and proximal tubule as an autocrine or paracrine regulator of hemodynamics and phosphate transport. PMID- 9227637 TI - Acid-base changes alter Mg2+ uptake in mouse distal convoluted tubule cells. AB - Metabolic alkalosis leads to renal magnesium conservation, whereas metabolic acidosis is associated with urinary magnesium wasting. Micropuncture studies suggest that these actions affect magnesium transport in the distal tubule. The cellular mechanisms of acid-base changes were investigated in an immortalized mouse distal convoluted tubule (MDCT) cell line. Intracellular free Mg2+ concentration ([Mg2+]i) was determined by microfluorescence using the Mg(2+) responsive dye, mag-fura 2. Mg2+ transport was assessed as a function of change in [Mg2+]i with time following placement of Mg(2+)-depleted cells into a buffer containing 1.5 mM magnesium. The uptake rate of Mg2+, d([Mg2+]i)/dt, into Mg(2+) depleted cells determined with a buffer solution of pH 7.4 was 178 +/- 21 nM/s. Mg2+ uptake at pH 8.0 was markedly increased 278 +/- 35 nM/s, whereas transport at pH 6.0 was significantly reduced to 121 +/- 15 nM/s. Mg2+ uptake at pH 7.4 was not stimulated with 20 or 40 mM bicarbonate, nor were the differences in Mg2+ uptake with pH associated with changes in membrane voltage. Mg2+ uptake was stimulated with membrane hyperpolarization at pH 6.0 but not at pH 8.0. Chlorothiazide (10(-4) M), which stimulates Mg2+ uptake by hyperpolarizing the membrane voltage, increased uptake at pH 6.0, 59 +/- 14%, but decreased it at alkaline pH of 8.0, -55 +/- 3%. Accordingly, MDCT cells become refractory to the stimulating effects of hyperpolarization at alkaline pH values. These studies show that protons may directly affect Mg2+ transport in MDCT cells. PMID- 9227638 TI - Endothelial cells protect vascular smooth muscle cells from H2O2 attack. AB - Endothelial-dependent vascular responses are altered in ischemic acute renal failure. Oxidants formed during reperfusion of ischemic kidneys injure the renal microvasculature and prevent recovery of renal function. To determine whether endothelial cells (EC) modulate oxidant attack on vascular smooth muscle cells (VSMC), rat mesenteric artery VSMC were grown on coverslips and then coincubated with bovine pulmonary artery EC grown in wells. In the absence of EC, H2O2 caused time- and concentration-dependent increases in VSMC injury as indicated by release of [3H]adenine. In contrast, addition of EC reduced H2O2-mediated (5 mM, 1 h) VSMC adenine release from 63.8 +/- 4.5% to 28.6 +/- 2.9% (P < 0.001). The protective effect of EC did not occur when H2O2 was added to the surface of VSMC unopposed to EC and was partially reversed when EC were treated with aminotriazole to inactivate catalase (41.7 +/- 2.7%). To determine whether EC nitric oxide (NO) modified H2O2 attack on VSMC, EC were treated with N omega nitro-L-arginine (L-NNA). The protective effect of EC was partially abrogated with L-NNA (53.8 +/- 4.3%). Treatment of EC with interleukin-1 beta (IL-1 beta) for 24 h prior to coincubation with VSMC enhanced the protective effect of EC. IL 1 beta-induced protection was reversed with L-NNA. No protection was observed when VSMC were treated with 8-bromoguanosine 3',5'-cyclic monophosphate, forskolin, or phorbol 12-myristate 13-acetate. Our conclusions are as follows. VSMC are protected by EC from luminal but not contraluminal oxidant attack. The protective effect of EC is mediated by catalase- and NO-dependent inactivation of oxidants. EC dysfunction could account for renal injury caused by oxidants formed during reperfusion of ischemic kidneys. PMID- 9227639 TI - Decreased potassium stimulates bone resorption. AB - Metabolic acidosis induces net calcium efflux (JCa+) from cultured bone, in part, through an increase in osteoclastic resorption and a decrease in osteoblastic formation. In humans provision of base as potassium (K+) citrate, but not sodium (Na+) citrate, reduces urine Ca (UCa), and oral KHCO3 decreases bone resorption and UCa in postmenopausal women. Potassium deprivation alone leads to an increase in UCa. To determine whether decreased extracellular K+ concentration ([K+]) at a constant pH, PCO2, and [HCO-3] alters JCa+ and bone cell activity, we measured JCa+, osteoblastic collagen synthesis, and osteoclastic beta-glucuronidase release from neonatal mouse calvariae cultured for 48 h in medium of varying [K+]. Calvariae were cultured in control medium (approximately 4 mM [K+]) or medium with mildly low K+ (MLK, approximately 3 mM [K+]), very low K+ (VLK, approximately 2 mM [K+]), or extremely low K+ (ELK, approximately 1 mM [K+]) (n > or = 9 in each group). Compared with control, ELK, but not MLK or VLK, resulted in a marked increase in JCa+ and an increase in beta-glucuronidase release and a decrease in collagen synthesis. JCa+ was correlated directly with medium beta glucuronidase activity and inversely with collagen synthesis. To determine whether the reduction in medium [K+] was associated with a decrease in intracellular pH (pHi), we measured pHi in MC3T3-E1 cells, a mouse osteoblastic cell line. Incubation in 1 mM [K+] led to a significant decrease in pHi compared with 3 mM [K+]. Thus incubation in a reduced [K+] medium stimulates JCa+ and osteoclastic enzyme release and inhibits osteoblastic collagen synthesis, which may be mediated by a reduction in bone cell pH. PMID- 9227640 TI - Parathyroid hormone inhibits Na(+)-K(+)-ATPase through Gq/G11 and the calcium independent phospholipase A2. AB - Previous studies from this laboratory have demonstrated that the 3-34 analog of parathyroid hormone (PTH) causes a 15-30% inhibition of Na(+)-K(+) adenosinetriphosphatase (Na(+)-K(+)-ATPase) activity in rat renal proximal tubules through the generation of an increase in intracellular arachidonic acid, followed by its conversion to 20-hydroxyeicosatetraenoic acid (20-HETE) [C. P. Ribeiro and L. J. Mandel. Am. J. Physiol. 262 (Renal Fluid Electrolyte Physiol. 31): F209-F216, 1992; and C. P. Ribeiro, G. Dubay, J. R. Falk, and L. J. Mandel. Am. J. Physiol. 266 (Renal Fluid Electrolyte Physiol. 35): F497-F505, 1994]. The present study also uses proximal tubule suspensions to further elucidate this signaling pathway. Guanosine 5'-O-(2-thiodiphosphate), 500 microM, an inhibitor of heterotrimeric GTP-binding proteins (G proteins), and an anti-Gq/G11 antibody (1:500) both blocked the inhibition of the Na(+)-K(+)-ATPase by PTH-(3-34). Furthermore, a 42-kDa protein was identified in proximal tubules by the anti Gq/G11 antibody (1:1,000). Bromoenol lactone (BEL), 1 microM, a suicide inhibitor of the calcium-independent 40-kDa phospholipase A2 (PLA2), prevented PTH-(3-34) inhibition of the Na(+)-K(+)-ATPase, unless exogenous 10 microM 20-HETE was added. In addition, BEL blocked the PTH-(3-34)-induced increase in arachidonic acid release in the proximal tubules. We conclude that a member of the Gq family and the calcium-independent 40-kDa PLA2 participate in the PTH-(3-34) signaling pathway in rat proximal tubules by mediating the steps between the binding of PTH (3-34) to its receptor and the subsequent generation of arachidonic acid. PMID- 9227641 TI - Antioxidant expression in experimental hydronephrosis: role of mechanical stretch and growth factors. AB - We assessed whether levels of renal reactive oxygen species (ROS) and antioxidant enzymes are perturbed in rats following unilateral ureteral obstruction (UUO). The mechanism of catalase perturbation was investigated using proximal tubule suspensions following stimulation with transforming growth factor (TGF)-beta and interleukin (IL)-1 and in a proximal tubular cell line (OKC) subjected to cyclic mechanical stretch, which mimics the early hydrodynamic derangement after UUO. Levels of catalase and copperzinc superoxide dismutase (Cu,Zn-SOD) mRNA from 96-h UUO rats showed a 5.5-fold (P < 0.001) and 5.0-fold (P < 0.001) decrease, respectively, compared with the contralateral unobstructed kidney (CUK). Levels of superoxide anion and hydrogen peroxide showed a significant 1.8-fold (P < 0.0001) and 14.0-fold (P < 0.0001) increase, respectively, in 96-h UUO kidney slice cultures. In situ hybridization and immunohistochemistry showed Cu,Zn-SOD and catalase mRNA and protein transcription expressed in proximal tubules of UUO and CUK specimens. Catalase mRNA levels were markedly downregulated following a 1 h exposure of isolated proximal tubules to TGF-beta (0.1-10 ng) and IL-1 (1-5 ng), in comparison to control proximal tubular suspensions. OKC subjected to cyclic mechanical stretch for 1-24 h had marked decrements in catalase mRNA levels, compared with unstretched cells at the same time point. These results indicate that a primary downregulation of proximal tubular Cu,Zn-SOD and catalase expression develops in the proximal tubules of UUO with consequent increments in cortical oxidant levels. These findings suggest that either an early mechanical disturbance produced by UUO or local tubular generation of cytokines can reduce tubular catalase expression. The downregulation of catalase mRNA expression, together with increased oxidant stress in the rat renal cortex post-UUO, may amplify the proinflammatory state of experimental hydronephrosis culminating in tubulointerstitial injury and fibrosis. PMID- 9227642 TI - ANG II-dependent HCO3- reabsorption in surviving rat distal tubules: expression/activation of H(+)-ATPase. AB - Distal tubules (DT) from sham or five-sixths nephrectomized (Nx) rats were perfused in vivo to evaluate the hypothesis that, after Nx, endogenous angiotensin II (ANG II) modulates DT in vivo bicarbonate reabsorption (JtCO2) via H(+)-adenosinetriphosphatase (H(+)-ATPase) and Na+/H+ exchange. In Nx rats JtCO2 was increased (65 +/- 7 vs. -24 +/- 21 pmol.min-1.mm-1, P < 0.01). Both luminal and intravenous AT1-receptor blockade by losartan reduced Nx DT JtCO2 (41 +/- 6 and 34 +/- 4 pmol.min-1.min-1, respectively, P < 0.05), whereas neither 10(-9) M nor 10(-11) M ANG II luminal perfusion increased JtCO2, suggesting preexisting high endogenous ANG II levels. The Na+/H+ antiporter inhibitors 5-(N-ethyl-N isopropyl)-amiloride and 5-(N,N-dimethyl)-amiloride were without effect. Luminal perfusion of 5 nM concanamycin A, a V-type H(+)-ATPase inhibitor, reduced Nx DT JtCO2 (45 +/- 8 pmol.min-1.mm-1, P < 0.05). In Nx A-type intercalated cells, we demonstrated cellular hypertrophy, elaboration of apical microplicae, and enhanced expression/apical polarization of H(+)-ATPase. Thus ANG II is an important determinant in sustaining brisk DT JtCO2 following Nx and is associated with enhanced expression and A-type intercalated cell apical polarization of H(+) ATPase. PMID- 9227643 TI - Chronic hypoxia induces LLC-PK1 cell proliferation and dedifferentiation by the activation of protein kinase C. AB - The effect of chronic hypoxia on the proliferation and dedifferentiation of LLC PK1 cells was examined. Cultures were exposed either to hypoxia (3% O2) or normoxia (18% O2), and [3H]thymidine incorporation, cell number, and sodium dependent glucose (Na/Glc) uptakes were assessed. Cultures exposed to hypoxia for 16 h significantly increased [3H]thymidine incorporation followed by a significant increase in cell number both at 24 and 48 h in comparison with respective normoxic controls. Cultures exposed to 24 and 72 h of hypoxia exhibited significant inhibition of Na/Glc uptake when compared with their respective normoxic counterparts. Significant inhibition of cell ATP levels were observed under hypoxic conditions. Acute reoxygenation of hypoxic cells normalized cell ATP levels without any effect on the Na/Glc uptake. Hypoxia also activated protein kinase C (PKC) at 1 and 4 h followed by a subsequent return to baseline with reactivation at 24 h, which remained sustained up to 72 h, suggesting both acute and sustained activation of PKC. Furthermore, the hypoxia induced alterations in [3H]thymidine incorporation as well as Na/Glc uptake were mitigated by inhibitors of PKC. These results indicate that chronic hypoxia induces both proliferation and dedifferentiation of LLC-PK1 cells mediated, in part, by the activation of PKC. PMID- 9227644 TI - Protein kinase A phosphorylation is involved in regulated exocytosis of aquaporin 2 in transfected LLC-PK1 cells. AB - Vasopressin-dependent translocation of aquaporin-2 (AQP2) between intracellular vesicles and the plasma membrane has been demonstrated in vivo and in vitro. Furthermore, the vasopressin-induced increase in apical membrane water permeability of renal principal cells is dependent on a rise in intracellular adenosine 3',5'-cyclic monophosphate and activation of protein kinase A (PKA). To determine whether trafficking of AQP2 is dependent on PKA phosphorylation, we first examined the effect of the PKA-inhibitor N-(2[[3-(4-bromophenyl)-2 propenyl]-amino]-ethyl)-5-isoquinolinesulfonam ide (H-89) on AQP2 translocation in transfected LLC-PK1 cells. Vasopressin-induced membrane insertion of AQP2 was completely inhibited by pretreatment of the cells for 60 min with H-89. This reagent also caused a dense accumulation of AQP2 in the Golgi region. Next, LLC PK1 cells were stably transfected with AQP2 cDNA in which the PKA phosphorylation site, Ser256, was replaced with alanine (S256A). S256A-AQP2 was not phosphorylated in vitro by PKA, and S256A-AQP2 was mainly localized to intracellular vesicles in the basal condition, similar to wild-type AQP2. However, after stimulation with vasopressin or forskolin, the cellular distribution of S256A-AQP2 remained unchanged. In addition, the usual vasopressin induced increase in endocytosis seen in AQP2-transfected cells was not observed in S256A-AQP2-transfected cells. These results demonstrate that the Ser256 PKA phosphorylation site is possibly involved in the vasopressin-induced trafficking of AQP2 from intracellular vesicles to the plasma membrane and in the subsequent stimulation of endocytosis. PMID- 9227645 TI - Use of World Wide Web server and browser software to support a first-year medical physiology course. AB - We describe the use of a World Wide Web (Web) server to support a team-taught physiology course for first-year medical students. Our objectives were to reduce the number of formal lecture hours and enhance student enthusiasm by using more multimedia materials and creating opportunities for interactive learning. On-line course materials, consisting of administrative documents, lecture notes, animations, digital movies, practice tests, and grade reports, were placed on a departmental computer with an Internet connection. Students used Web browsers to access on-line materials from a variety of computing platforms on campus, at home, and at remote sites. To assess use of the materials and their effectiveness, we analyzed 1) log files from the server, and 2) the results of a written course evaluation completed by all students. Lecture notes and practice tests were the most-used documents. The students' evaluations indicated that computer use in class made the lecture material more interesting, while the on line documents helped reinforce lecture materials and the textbook. We conclude that the effectiveness of on-line materials depends on several different factors, including 1) the number of instructors that provide materials; 2) the quantity of other materials handed out; 3) the degree to which computer use is demonstrated in class and integrated into lectures; and 4) the ease with which students can access the materials. Finally, we propose that additional implementation of Internet-based resources beyond what we have described would further enhance a physiology course for first-year medical students. PMID- 9227646 TI - A simple analog of visual field retinal projection. AB - Basic biophysical principles of the eye, such as the retinal projection of the visual field, bidimensional image inversion, blind point determination, visual field and retinal quadrants, perimetric principles, and determination of scotoma, are seen on a working anatomic model of the eye and its visual field that can easily be constructed at very low cost and from easily obtained materials: an opaque glass globe, a styrofoam hemisphere, a wood screen, bulbs, sockets, wire, and switches. This model has been used for many years in our undergraduate medical physiology courses and has replaced the classical model of candle image inversion by a converging lens. PMID- 9227647 TI - A quarter-long exercise that introduces general education students to neurophysiology and scientific writing. AB - Providing large numbers of general education students with an introduction to science is a challenge. To meet this challenge, a quarter-long neurophysiology project was developed for use in an introductory biology course. The primary goals of this multistep project were to introduce students to the scientific method, scientific writing, on-line scientific bibliographic databases, and the scientific literature, while improving their academic literacy skills. Students began by collecting data on their own circadian rhythms in autonomic, motor, and cognitive function, reliably demonstrating the predicted circadian changes in heart rate, eye-hand coordination, and adding speed. Students wrote a journal style article using pooled class data. Students were prepared to write the paper by several methods that were designed to improve academic language skills, including a library training exercise, "modeling" of the writing assignment, and drafting of subsections of the paper. This multistep neurophysiology project represents a significant commitment of time by both students and instructors, but produces a valuable finished product and ideally gives introductory students a positive first experience with science. PMID- 9227648 TI - Modeling blood flow in vessels with changeable caliber for physiology and biophysics courses. AB - A model based on elementary principles of hydrodynamics and mathematics is proposed for classroom research on concepts related to blood flow physiology. This is an analog model of the vascular system in which blood flow is represented by electrical current flowing in a resistance circuit. The model permits analysis of the change in hemodynamics with local stenosis of both large and peripheral vessels. PMID- 9227649 TI - Philosophy of science and physiology education. AB - Since the mid-1960s, philosophy of science [particularly that derived from Kuhn's work (The Structure of Scientific Revolutions, Chicago: University of Chicago Press, 1962)] has become an informal part of the education of scientists worldwide, including physiologists. However, recent postmodernist developments have enraged a number of scientists, who would like to sever any ties with philosophy of science. The author contends that the perceived conflict is due mainly to a misunderstanding of the implications of constructivist assertions and partially to flawed reasoning in a few constructivist approaches. There is no fundamental conflict that would justify the elimination of philosophy of science from science education. PMID- 9227650 TI - A new PhD training track: a proposal to improve basic science teaching. AB - There has been increasing criticism of medical basic science teaching; much of this has focused on overcrowding of the curriculum, inadequate application to clinical medicine, and the limited commitment of the faculty to teach. We have analyzed some of the factors that may contribute to these complaints, such as the fragmentation of physiology and the conflicting roles of the medical basic scientist. We have also reviewed some previous suggestions for improving basic science teaching. We suggest that a basic scientist with a background of integrative physiology, pharmacology, anatomy, and pathology, with a special emphasis on pathophysiology, would be well qualified to assume an important role in the medical education of the future. Because there is at present no established training program of this type, we have proposed a PhD training track with this objective and have listed some of the advantages and disadvantages of such a program. PMID- 9227651 TI - Science in developing countries: whose agenda? PMID- 9227652 TI - Teaching versus research in academia. PMID- 9227653 TI - Cryptosporidiosis and public health. PMID- 9227654 TI - Case in point. Chronic aortic dissection. PMID- 9227655 TI - Left bundle branch block with ST segment elevation. PMID- 9227656 TI - Case in point. Bilateral cerebellar infarcts in antiphospholipid syndrome. PMID- 9227657 TI - Acute onset of dermatitis, hepatitis, and eosinophilia. AB - A 48-year-old man presented to the emergency department complaining of fever, chills, myalgias, and diffuse abdominal discomfort of four days' duration. A nonpruritic rash had developed on his left palm, arms, legs, and buttocks on the fourth day. He had not had respiratory symptoms, nausea, vomiting, or diarrhea. PMID- 9227658 TI - Alzheimer's disease: the genetics of risk. AB - Each copy of an epsilon 4 allele of the gene encoding the lipid carrier apoE increases the probability of the disease and shifts its onset to lower ages. Yet the gene does not cause the disease. Thus, although genotyping is highly valuable in diagnosis and drug testing, it cannot offer useful predictions about persons whose cognition is unimpaired. After discovery of a second susceptibility gene, predictability will change profoundly. PMID- 9227659 TI - Management after exposure to tuberculosis. AB - The increased incidence of tuberculosis-coupled with the emergence of mycobacterial strains resistant to the most effective drugs-has highlighted the importance of identifying transmission and preventing active disease. Skin test conversion can document infection, except in most patients vaccinated with bacille Calmette-Guerin. Prophylactic medication is effective, but not without complications. PMID- 9227660 TI - Respiratory syncytial virus: not just for kids. AB - Respiratory syncytial virus (RSV) earned its reputation as a pediatric pathogen, especially in children under two years of age. RSV infection in adults was considered a significant problem only for certain high-risk populations. Evidence now indicates that the infection occurs frequently in previously healthy adults. Clinical presentation varies greatly. PMID- 9227662 TI - Acute care costs of the oldest old. AB - Caring for elderly patients in the hospital costs less as they age. Among the reasons: Their ailments are less expensive to treat, they receive less aggressive care, and they are treated in less expensive hospitals. PMID- 9227661 TI - Regulation of appetite and body weight. AB - Short-term variations in caloric intake and energy expenditure-including attempts by obese patients to lose weight-tend to be modified by the body's long-term weight regulatory system. Hormones such as leptin and insulin participate in this system, which links changes in body fat content to appropriate compensatory responses in the hypothalamus. Correction of defects in the system might permit sustained weight loss in obese patients. PMID- 9227663 TI - Respiratory distress in a former drug abuser. PMID- 9227664 TI - Recognizing depression in primary care patients. PMID- 9227665 TI - Ask the right questions if you plan to go from solo to group. PMID- 9227666 TI - Sudden retrosternal pain in a young weight lifter. PMID- 9227667 TI - A diabetic man with persistent joint pain and infection. PMID- 9227668 TI - Selection of an antidepressant: mirtazapine. AB - The number of antidepressants has expanded dramatically in the last decade as a result of the ability to design psychiatric medications in a rational manner. This paper provides a broad overview of these options while focusing on the most recently approved antidepressant, mirtazapine. The paper discusses the five major features that a physician needs to consider when selecting a medication for a patient: safety, tolerability, efficacy, payment, and simplicity. These five features are summarized by the mnemonic STEPS. The paper also proposes a pharmacologically based classificatory system that divides current antidepressants into eight groups based on the mechanism of action thought to underlie their antidepressant efficacy. Such a classificatory system can serve as an organizing principle for the practicing physician. PMID- 9227669 TI - Antidepressants. PMID- 9227670 TI - Safety and tolerability of the new antidepressants. AB - The newer serotonergic antidepressants have become popular particularly because of improved tolerability and safety. The evidence supporting this belief is reviewed. The factors that contribute to the development of somatic symptoms are examined, including the depression itself and the drugs used for treatment. The rates for individual side effects with the serotonin selective reuptake inhibitors, nefazodone, and venlafaxine are presented and compared with the adverse event experience for mirtazapine. PMID- 9227671 TI - Efficacy issues with antidepressants. AB - Clinical trials of antidepressant medications have shown that, overall, these drugs are effective, as measured by a > or = 50% decrease in Hamilton Rating Scale for Depression (HAM-D) total scores in about two thirds of patients. However, the results of long-term trials under rigorously controlled conditions show that, even with close follow-up and provision of interpersonal psychotherapy, a third or more of the patients will not achieve or maintain a response to medication for depression. Nevertheless, the improved efficacy of some antidepressants for certain features or types of depression has been shown. Factors associated with a better response to a specific agent or class of drugs include severity of symptoms, patient age, and the symptom profile of the depressive episode, as revealed by assessment scales or subscale scores for selected symptoms. Moreover, a number of studies indicate that a patient's early response to a given medication may assist in predicting long-term outcome. However, outcome measures in traditional trials of antidepressant drug efficacy, such as a 50% reduction in scores on one or more depression rating scales, do not necessarily reflect an improvement in the patient's ability to function in the workplace; they only show that a particular patient at a particular time has responded to treatment in a significant manner by measurement of a depression scale. PMID- 9227672 TI - Development of new antidepressants. AB - A large number of novel antidepressants acting on a variety of neurotransmitter receptors are currently undergoing clinical evaluation. Most agents have a dual mechanism of action on two or more neurotransmitter receptors, including two serotonin receptors, two noradrenergic receptors, or a combination of serotonin and noradrenergic mechanisms. The most recently approved agent, mirtazapine, is an example of this approach of simultaneously targeting both the serotonergic and noradrenergic systems. Specifically, mirtazapine's alpha 2 antagonism disinhibits both serotonin and norepinephrine neurotransmission while its serotonin-2 and serotonin-3 antagonist properties reduce the side effects normally associated with nonselective serotonin receptor activation by serotonin selective reuptake inhibitors (SSRIs). This approach of "designer polypharmacy" applies principles of rational pharmacologic combinations to enhance efficacy and improve tolerability of the new and emerging antidepressants. PMID- 9227673 TI - Intercenter agreement in reading Doppler embolic signals. A multicenter international study. AB - BACKGROUND AND PURPOSE: Different frequencies of asymptomatic Doppler embolic signals have been reported in studies. There has been concern that different criteria for identification may account for some of this variation. A previous reproducibility study between two centers found good agreement, but no studies among large numbers of centers have been performed. We performed an international reproducibility study among nine centers, each of which had published recent studies of embolic signal detection in peer-reviewed journals. METHODS: Each center performed blinded analysis of a taped audio Doppler signal composed of transcranial Doppler middle cerebral artery recordings from 6 patients with symptomatic carotid artery stenosis. The exact time of any embolic signal was recorded. Six centers also measured the intensity increase of any embolic signals detected. Interobserver agreement was determined by a method based on the proportion of specific agreement. RESULTS: Seven centers reported between 39 and 55 signals, but one center reported 142 embolic signals. The probability of agreement between observers was .678, which rose to .791 when the data from the highest reporting center were excluded. Introducing a decibel threshold resulted in a significant increase in the probability of agreement; a decibel threshold of > 7 dB resulted in a probability of agreement of .902. Intensity measurements made by different centers were usually highly correlated, but this was not always the case, and 3 of the 15 correlations were not significant. The absolute values of the intensities measured varied between centers by as much as 40%. CONCLUSIONS: Although most centers report similar numbers of embolic signals, some use less specific criteria and report more events. The use of a decibel threshold improves reproducibility. However, intensity thresholds developed by one center cannot be directly transferred without validation to another center; differing methods of measurement are being used, and this results in different intensity values for the same embolic signals, even when the same equipment is used. PMID- 9227675 TI - Cerebral microembolism and early recurrent cerebral or retinal ischemic events. AB - BACKGROUND AND PURPOSE: We investigated whether cerebral microembolism as detected by transcranial Doppler ultrasonography (TCD) identifies patients at an increased risk for early, recurrent cerebral or retinal ischemic events. METHODS: Records of consecutive patients examined during a 40-month period in the Neurovascular Laboratory were reviewed for the presence of cerebral microembolism. Of the original 302 patients, 229 with 310 arteries met inclusionary criteria. Follow-up information was obtained from the laboratory's database as well as the hospital records. Microembolus detection studies were performed on TC-2000 or TC-2020 instruments equipped with special software, and criteria established a priori were used for microembolus selection. TCD testing was performed a median interval of 9 days after the initial symptoms of cerebral ischemia. Severity of arterial stenosis was determined by cerebral angiography or noninvasive methods. RESULTS: Microembolic signals were detected more frequently in symptomatic (40/140; 28.6%) than asymptomatic (21/170; 12.4%) arteries (P < .001). Ten recurrent ischemic events occurred during a median follow-up of 8 days after TCD examination, all in the territories of symptomatic arteries. Nine events occurred in the territories of microembolic signal positive arteries (9/61; 14.8%) and one in the territory of a microembolic signal-negative artery (1/249; 0.4%) (P < .00). No association was detected in the subgroup with known cardiac lesions. Microembolic signals were more frequent in arteries with lesions causing 70% or more stenosis or occlusion (26/99; 26.3%) than in those with a degree of stenosis less than 70% (17/126; 13.5%) (P = .016). CONCLUSIONS: In this retrospective study, microembolic signals were more common in the territories of symptomatic arteries and particularly those with severely stenotic lesions. During a short follow-up, recurrent ischemic events were more common along the territories of arteries with TCD-detected microembolism and previous symptoms of cerebral or retinal ischemia. PMID- 9227674 TI - Microembolic signals with serial transcranial Doppler monitoring in acute focal ischemic deficit. A local phenomenon? AB - BACKGROUND AND PURPOSE: The occurrence of microembolic signals (MES) in patients with transient ischemic attack (TIA) or stroke has already been described, but the diagnostic and prognostic value of this finding is still debated. METHODS: We evaluated 90 consecutive patients admitted for their first hemispheric TIA or ischemic stroke within 72 hours of onset. All of them underwent 30-minute bilateral transcranial Doppler monitoring of middle cerebral arteries, within 72 hours of onset. The monitoring was repeated after an additional 24 hours and after 7 days. We then classified the episodes in the following etiologic categories: cardioembolic, atherothrombotic, small-vessel disease, mixed cases, unknown origin, and other causes. RESULTS: We included 75 patients, with a mean interval of registration of 32.04 +/- 19.39 hours. There were 9 patients with MES (12%). All MES were recorded only on the symptomatic middle cerebral artery, and the majority were recorded during the first or the second registration. No statistically significant difference was found in risk factors and hematologic parameters. Five patients (56%) had atherothrombotic episodes, 3 patients (33%) had cardioembolic episodes, and 1 patient (11%) had a protein S deficit. No patient with MES had small-vessel disease (P = .01). CONCLUSIONS: MES are an infrequent finding in patients with TIA or ischemic stroke within 72 hours of onset, but they can be recorded more easily with serial registration. In our patients, MES were found only on the symptomatic middle cerebral artery and were present in atherothrombotic and cardioembolic episodes but not in small-vessel disease. PMID- 9227676 TI - Transoccipital power-based color-coded duplex sonography of cerebral sinuses and veins. AB - BACKGROUND AND PURPOSE: Power-based transcranial color-coded duplex sonography is a new development for cerebrovascular imaging that is suited for detection of slow velocities. The purpose of this study was to evaluate the ability of this technique to detect cerebral sinuses and veins by means of the occipital window and to provide reference data. METHODS: The straight and inferior sagittal sinuses, great and internal cerebral veins, and basal veins were insonated in 120 normal subjects. The number of identified vessels, peak systolic (PSV) and end diastolic (PDV) velocities, and resistance indices were determined. RESULTS: In subjects aged 20 to 59 years, straight sinuses were identified in 81% and great and internal cerebral veins in 34%. In subjects aged 60 to 79 years, straight sinuses were detected in 50%, great cerebral veins in 20%, and internal cerebral veins in 13%. All insonated inferior sagittal sinuses and basal veins were missed. Velocities were highest in straight sinuses (PSV, 35 [7 to 64] cm/s; PDV, 23 [2 to 43] cm/s), slower in great cerebral veins (PSV, 23 [12 to 34] cm/s; PDV, 16 [7 to 26] cm/s), and slowest in internal cerebral veins (PSV, 14 [10 to 18] cm/s; PDV, 10 [5 to 15] cm/s) (mean with 95% confidence intervals [CIs]). Straight sinus velocities decreased with age for PSV (20 to 39 years, 40 [7 to 73] cm/s; 60 to 79 years, 28 [9 to 46] cm/s; P < .01) and PDV values (20 to 39 years, 28 [4 to 52] cm/s; 60 to 79 years, 16 [5 to 26] cm/s; P < .001) (mean with 95% CIs) and were higher in women than men in the group aged 20 to 39 years. (P < .05). Resistance indices increased with age in the straight sinus (20 to 39 years, 0.30 [0.18 to 0.42]; 60 to 79 years, 0.42 [0.31 to 0.53]; P < .001) (mean with 95% CIs). CONCLUSIONS: Transoccipital power-based color-coded duplex sonography enabled imaging and velocity measurements in the straight sinus of subjects aged 20 to 59 years. In elder subjects detection rate of the straight sinus decreased, and it was low for deep cerebral veins in all age groups. PMID- 9227678 TI - Doppler microembolic signals in children with prosthetic cardiac valves. AB - BACKGROUND AND PURPOSE: The aim of this study was the evaluation of the prevalence and counts of Doppler microembolic signals (MES) in children with prosthetic cardiac valves and their comparison to those obtained in corresponding adult patients. PATIENTS AND METHODS: Nine children and 43 adults with ATS valves implanted in the aortic position were monitored over both middle cerebral arteries with transcranial Doppler ultrasound. MES were identified on-line according to standard criteria. Heart rate and rhythm, valve type, size and duration, patients' height, International Normalized Ratio, and prevalence of neurological complications were obtained from all study participants. RESULTS: MES prevalence and counts were significantly higher in children compared with adult patients (100% versus 25.5% and 58 [18.5 to 115.5] versus 55 [2 to 10.5], median, 95% CI, respectively). No corresponding differences in valve duration of valve implant were evident, but children has heart rates and were significantly smaller compared with adults. A positive correlation between patients' size, heart rate, and MES counts was noted. CONCLUSIONS: MES counts in children with mechanical prosthetic valves are significantly higher compared with those in corresponding adults. We hypothesize that this is due to (1) the shorter distance between aortic valve and middle cerebral artery, since cavitation bubbles have a short life span and are bound to dissolve with time, and (2) the faster heart rate in children, resulting in a higher number of valve closures per minute. PMID- 9227677 TI - Do chronic middle cerebral artery stenoses represent an embolic focus? A multirange transcranial Doppler study. AB - BACKGROUND AND PURPOSE: It remains uncertain whether the annual stroke risk of 7% to 8% in middle cerebral artery (MCA) stenosis is of embolic or hemodynamic origin. Preliminary reports provide evidence of emboli exiting from acute MCA stenoses, detected by transcranial Doppler (TCD) sonography. With multirange monitoring before and after the stenosis, TCD monitoring may help for the first time to differentiate microemboli exiting from the MCA stenosis from those with a source proximal to the MCA stenosis. We searched for microembolic signals (MES) using multigated monitoring in patients with chronic MCA stenoses. METHODS: Fifty eight patients with 78 chronic stenoses of the MCA were enrolled in the study. Additional sources of embolism were ruled out by extensive clinical workup. Twenty-four patients were treated with coumarin, whereas 28 patients received aspirin. The remaining 6 patients discontinued their medication after a few weeks. The sample volume of the multirange probe was placed on either side of the stenotic area of the MCA. RESULTS: Twenty-three (29.5%) of the stenoses were low grade, 18 (23%) were moderate, and 37 (47.5%) were severe. Thirty-seven (47%) of the stenoses were symptomatic and 41 (53%) were asymptomatic before study entry. During follow-up, 2 strokes and 7 transient ischemic attacks occurred. Computer tomography revealed two watershed-type infarcts. Sufficient insonation of the prestenotic and poststenotic segments of the MCA was possible in 70 stenoses (90%). No MES could be detected during a total of 1740 minutes' monitoring time distal to the MCA stenoses, regardless of the patients' medication. MES were also absent in the contralateral MCA. CONCLUSIONS: MES are not detectable in patients with chronic MCA stenoses of different degrees. No MES were found in either symptomatic or asymptomatic stenoses, regardless of the patients' medication. These results indicate that chronic MCA stenoses do not represent a significant embolic source. The absence of MES in the prestenotic Doppler sample volume, the watershed-type infarcts during follow-up, and the absence of small-vessel disease on computed tomography suggests that hemodynamic mechanisms are responsible for recurrent cerebral ischemia. PMID- 9227679 TI - Assessment of the efficacy of noninvasive screening for patients with asymptomatic neck bruits. AB - BACKGROUND AND PURPOSE: Several recent clinical trials have shown that endarterectomy is efficacious in patients with asymptomatic carotid artery stenosis. The purpose of this study was to evaluate the effectiveness of various test strategies for screening and diagnosing carotid artery disease. METHODS: We constructed a model of the natural history of carotid artery disease using literature-based estimates of the prevalence and incidence of carotid artery stenosis and associated morbidity and mortality. Markov cohort simulation was used to estimate the mean quality-adjusted life years and monetary costs associated with various management strategies. RESULTS: Screening is cost effective in the baseline model. Key parameters affecting the efficacy of screening are prevalence of operable lesions, benefit of surgery, surgical complication rates, quality of life with stroke, rate of stenosis progression, and excess morbidity and mortality. CONCLUSIONS: Asymptomatic patients with carotid bruits may benefit from screening if the prevalence rate is > or = 20%, the benefits and risks associated with surgery are similar to those observed in the Asymptomatic Carotid Atherosclerosis Study, and the quality of life with stroke is considerably lower than the quality of life without stroke. Ultrasound followed by three-dimensional time-of-flight MR angiography, if indicated, is a promising test strategy. PMID- 9227680 TI - Impaired dynamic cerebral autoregulation in carotid artery stenosis. AB - BACKGROUND AND PURPOSE: If it could be determined whether cerebral blood flow can be maintained (autoregulated) during transient falls in arterial blood pressure, we might be able to identify patients with carotid stenosis who are at risk of stroke. However, conventional methods of determining autoregulation in such patients are invasive and/or expensive. METHODS: We used a new noninvasive method to estimate dynamic cerebral autoregulation in 27 patients with carotid stenosis and 21 age-matched normal controls. After a stepwise fall in arterial blood pressure, we determined the rate of rise of middle cerebral artery blood flow velocity compared with that of arterial blood pressure. We compared the method with a conventional method of determining cerebral hemodynamics, CO2 reactivity. RESULTS: Autoregulatory index (ARI) was significantly reduced in middle cerebral arteries ipsilateral to a stenosed/ occluded carotid artery: mean +/- SD 3.3 +/- 2.2 compared with normal controls (6.3 +/- 1.1; P < .0001) and nonstenosed carotid arteries in patients (5.9 +/- 2.1; P < .002). A subgroup of patients with severe impairment was identified. ARI returned to normal after carotid endarterectomy was performed. In a number of cases, ARI was impaired in the presence of CO2 reactivity. CONCLUSIONS: This simple technique allows identification of impaired autoregulation in patients with carotid artery disease. It may allow identification of patients at risk from transient falls of blood pressure as may occur at the onset of antihypertensive therapy and during surgery. It may allow a subgroup of patients with asymptomatic carotid stenosis who are at risk of hemodynamic stroke to be identified. PMID- 9227681 TI - Continuous intraoperative monitoring of middle cerebral artery blood flow velocities and electroencephalography during carotid endarterectomy. A comparison of the two methods to detect cerebral ischemia. AB - BACKGROUND AND PURPOSE: Intraoperative monitoring of brain function may influence the outcome of carotid endarterectomy (CEA). METHODS: We performed transcranial Doppler (TCD) monitoring of middle cerebral artery blood flow velocities (VMCAs) and eight-channel electroencephalographic (EEG) recording simultaneously in 82 patients undergoing CEA. Thiopental narcosis limited EEG interpretation in 11 patients, thus allowing direct comparison of both methods in 71 patients. RESULTS: There was a significant correlation between VMCA decrease and the frequency of EEG changes after carotid clamping (P < .001). Eight patients (11%) showed a VMCA decrease exceeding 60%, accompanied by EEG changes in 7 patients. Altogether, 16 patients (22%) showed severe or moderate EEG changes. Stenosis or occlusion of the contralateral carotid artery led to an increase of abnormal findings with both monitoring methods, which was, however, significant only for TCD (P < .05). Four patients (4.8%) suffered intraoperative transient ischemic attacks. In 3 of these patients, there were no abnormal findings with either of the methods. The events were thus unpredictable and probably of embolic origin. The fourth patient showed VMCA decrease to 0 and severe EEG changes. Nine patients had severe or moderate EEG changes without significant VMCA decrease and without complications. EEG monitoring alone in these would have led to unnecessary use of a shunt with the increased risk of embolism. CONCLUSION: EEG and TCD monitoring are complementary techniques. Their results showed a good overall correlation but with marked differences in the individual patient. TCD monitoring alone was sensitive enough to prevent ischemic intraoperative complications. EEG findings are of limited value when barbiturates are used. PMID- 9227682 TI - Impaired calcium regulation in subcortical vascular encephalopathy. AB - BACKGROUND AND PURPOSE: A number of clinical observations and first in vitro findings indicate that chronic cerebral ischemia influences immunologic status, such as the proliferative response of T lymphocytes. The purpose of the present report was to assess (1) whether changes of immune function are likewise detectable in patients with progressive subcortical vascular encephalopathy (SVE) by investigating the [Ca2+]i homeostasis of lymphocytes and (2) whether differences exist in calcium regulation between lymphocytes from SVE patients and from Alzheimer's disease (AD) patients. This is of great interest, since specific changes have been reported recently in AD patients. METHODS: [Ca2+]i was recorded in 26 patients with SVE, 26 age-matched nondemented control subjects, and 26 age matched patients with AD. Basal [Ca2+]i and [Ca2+]i after lymphocyte activation with the mitogen phytohemagglutinin (PHA) were measured with the fura 2 method. In addition, modulation of the Ca2+ signaling by the peptide beta-amyloid and the potassium channel blocker tetraethylammonium was studied. RESULTS: Basal [Ca2+]i was not different between patients and control subjects. After stimulation with PHA, however, a significant reduction of the Ca2+ response could be observed in lymphocytes of SVE patients compared with control subjects and with AD patients, providing evidence that the Ca2+ homeostasis of lymphocytes is impaired in SVE. The effect of the peptide beta-amyloid, the major constituent of senile plaques in AD brain, on Ca2+ signaling was similar in SVE patients and nondemented control subjects but typically reduced in cells of AD patients. Potassium channels were not involved in the impaired Ca2+ response of SVE lymphocytes after cell activation. CONCLUSIONS: [Ca2+]i is not only one of the most important second messengers in signal transduction of many cells but also an early event in the signal cascade of cell proliferation as a reaction to antigen recognition. This mechanism seems to be impaired in SVE. These findings may result in new insights regarding the pathogenesis of this disease and the possible involvement of inflammatory or immunologic disturbances. PMID- 9227683 TI - T2-weighted hyperintense MRI lesions in the pons in patients with atherosclerosis. Amsterdam Vascular Medicine Group. AB - BACKGROUND AND PURPOSE: Pontine hyperintense lesions (PHL) on T2-weighted MRI have been recognized recently. Histopathological findings resemble periventricular leukoaraiosis, and a vascular etiology has been suggested. We studied the frequency and the associated factors of PHL in patients with symptomatic atherosclerosis. METHODS: Two independent observers assessed brain MRIs in a prospective cohort of patients with symptomatic atherosclerosis. Only patients in whom both observers scored PHL on T2- and proton density-weighted images, but not on T1-weighted images, were considered to have the lesion. RESULTS: We studied 229 patients 31% presenting with ischemic stroke, 31% with myocardial infarction, and 38% with peripheral artery disease. Both observers scored PHL in 23% of all patients. Patients with PHL were significantly older and had more lacunar infarcts and periventricular leukoaraiosis than patients without PHL. There were more women, more hypercholesterolemic and diabetic patients, and more cortical infarcts on MRI (P = NS). After logistic regression the presence of leukoaraiosis (odds ratio, 2.4; 95% confidence interval, 1.6 to 3.4) and lacunar infarcts (odds ratio, 2.2, 95% confidence interval, 1.5 to 3.1) remained independently associated with PHL. PHL was more common in patients with ischemic strokes (39%) than in patients with myocardial infarctions (11%) or peripheral artery disease (19%) (P < .001). CONCLUSIONS: We found that PHL on T2- and proton density-weighted MR images are often found in patients with symptomatic atherosclerosis. The association with periventricular leukoaraiosis and lacunar infarcts suggests that PHL is a variant of leukoaraiosis, with possibly the same pathophysiology. The clinical symptoms and consequences of PHL, however, are not yet clear. PMID- 9227684 TI - Parental history of cardiovascular disease and risk of stroke. A prospective follow-up of 14371 middle-aged men and women in Finland. AB - BACKGROUND AND PURPOSE: Studies on risk factors for stroke have been less intensive than those for coronary disease. Only a few studies have addressed the question of the role of heredity in the occurrence of stroke. We analyzed whether a positive parental history of cardiovascular disease predicts the risk of stroke independently from other risk factors and whether the role of parental history varies by age and stroke subtypes. METHODS: This study was a prospective follow up of 14371 middle-aged men and women. A positive parental history of cardiovascular disease was defined as either stroke or coronary disease before the age of 60 years. The end point of the follow-up was an incident case of stroke. Multivariate analyses were performed with the Cox proportional hazards model. RESULTS: The risk ratio of stroke after multifactorial adjustment (age, smoking, blood pressure, cholesterol, diabetes, and education) associated with a positive parental history of stroke was 1.89 (P = .004) in men and 1.80 (P = .007) in women. The association between parental history of stroke and the risk of stroke was stronger among subjects aged 25 to 49 years than among older subjects. Parental history of coronary disease was not associated with the risk of stroke in men, but in women it had a borderline significant association with the risk of ischemic stroke. CONCLUSIONS: A positive parental history of stroke predicted the risk of stroke independently from the other risk factors. PMID- 9227685 TI - Stroke incidence and mortality correlated to stroke risk factors in the WHO MONICA Project. An ecological study of 18 populations. AB - BACKGROUND: The aim of the present study was to determine the extent to which the variation in conventional risk factors contributed to the variation in stroke incidence among these populations. METHODS: Within the WHO MONICA Project, stroke has been recorded in 18 populations in 11 countries. In population surveys, risk factors for cardiovascular diseases have been examined in the age group 35 to 64 years. Over a 3-year period, 12,224 acute strokes were registered in men and women within the same age range. RESULTS: The highest stroke attack rates were found in Novosibirsk in Siberia, Russia, and Finland, with a more than three-fold higher incidence than in Friuli, Italy. The mean diastolic blood pressure among the populations differed by 15 mm Hg between Novosibirsk (highest) and Denmark (lowest). In multiple regression analyses, the presence of conventional cardiovascular risk factors (smoking and elevated blood pressure) explained 21% of the variation in stroke incidence among the population in men and 42% in women. In Finland, in China, and in men in Lithuania, the stroke incidence rates were higher than expected from the population risk factor levels. CONCLUSION: Prevalence of smoking and elevated blood pressure explain a substantial proportion of the variation of stroke attack rates between populations. However, other risk factors for stroke that were not measured in the present study also contribute considerably to interpopulation differences in stroke rates. PMID- 9227686 TI - Cost of stroke in The Netherlands from a societal perspective. AB - BACKGROUND AND PURPOSE: Cerebrovascular disorders are associated with a high level of morbidity and mortality and call for considerable resources. The objective of this study was to determine from a societal perspective the medical consumption (direct costs) and productivity losses (indirect costs) caused by cerebrovascular disorders in the Netherlands. METHODS: This study can be characterized as a cost-of-illness study based on prevalence data. All data gathered refer to 1993. Cerebrovascular disorders are defined according to the International Classification of Diseases, 9th Revision (ICD-9) classification. Data from medical registrations and national statistics have been analyzed. For both direct and indirect costs, volume and cost components are presented. To test the likelihood of the assumptions, a sensitivity analysis was performed. RESULTS: The cost of cerebrovascular disorders in the Netherlands in 1993 amounted to 2.5 billion Dutch guilders, of which 1.9 billion were spent on medical consumption. It was found that direct costs are generated mainly by the long-term care of inpatients (nursing homes and hospitals). The productivity losses were relatively low in comparison with other diseases, probably due to the fact that most patients with cerebrovascular disorders are elderly. CONCLUSIONS: More than 3% of the Dutch annual healthcare budget is spent on patients suffering from cerebrovascular disorders. Costs in the future may be influenced by, among other things, demographic changes, new therapies, and cost-reduction programs introduced by the government. PMID- 9227687 TI - Trunk control test as an early predictor of stroke rehabilitation outcome. AB - BACKGROUND AND PURPOSE: The aim of this study was to investigate the construct and predictive validity of the Trunk Control Test (TCT) in postacute stroke patients by comparing TCT scores at admission and discharge with the Functional Independence Measure (FIM) scores. METHODS: Forty-nine patients participated in the study. The TCT examines four movements: rolling from a supine position to the weak side (T1) and to the strong side (T2), sitting up from a lying-down position (T3), and sitting balance (T4). The FIM is an 18-item scale (13 motor [motFIM] and 5 cognitive [cognFIM]) used to determine the level of dependence of patients in daily life. RESULTS: Thirty-six patients (73%) increased their TCT overall score at discharge. The TCT item-total correlations were high, both at admission and discharge (P < .0001). The individual TCT items were intercorrelated. Furthermore, the homogeneity of the TCT was confirmed by a high Cronbach's index. High correlations were found between admission and discharge scores in the different tests (TCT, FIM, and motFIM; P < .0001) and between TCT at admission and FIM (P < .0001) and motFIM (P < .0001) at admission. TCT at admission alone explained 71% of the variance in motFIM at discharge. CONCLUSIONS: The TCT showed a good sensitivity to change in assessing recovery of stroke patients. The high item-total correlation and Cronbach's alpha value of the TCT suggest that there is one homogeneous construct underlying the item list. The TCT construct validity was confirmed by the correlation between this test and the FIM scores. TCT at admission predicted motFIM at discharge even better than motFIM at admission alone. Possibly, the TCT captures basic motor skills that foreshadow the recovery of more complex behavioral skills described by the FIM. PMID- 9227688 TI - The influence of visual neglect on stroke rehabilitation. AB - BACKGROUND AND PURPOSE: The poor outcome observed in stroke patients with visual neglect may be due to greater stroke severity or nonspecialist management. METHODS: The effects of visual neglect were studied prospectively in 150 consecutive stroke patients with comparable stroke pathology and motor severity managed on a stroke unit. A randomized study was subsequently undertaken in 50 stroke patients with visual neglect to evaluate the effectiveness of spatial cueing during motor activity on functional outcome and resource use in these patients. RESULTS: Visual neglect was present in 47 (32%) of a selected group of 146 patients (mean age, 77.0 +/- 8.2 years; 42% men) with moderate stroke severity. There were no differences in demography, prestroke function, or motor power in the arm (2.6 +/- 1.7 versus 2.3 +/- 2.1) or the leg (3.2 +/- 1.4 versus 3.0 +/- 1.6) on the affected side compared with 99 patients with no visual neglect. Although patients with visual neglect had lower median initial (4 versus 5, P < .01) and discharge (14 versus 16, P < .01) Barthel Index scores, equal proportions of patients were discharged home (60% versus 65%) or to institutions (34% versus 33%) in both groups. The durations of hospitalization (64 versus 36 days, P < .001) and therapy input (47.7 versus 27.8 hours; P < .01), however, were significantly greater in patients with visual neglect. The randomized controlled study showed a trend toward higher Barthel scores at 12 weeks (14 versus 12.5, P = NS) and significant reduction in median length of hospital stay (42 versus 66 days) in patients receiving spatiomotor cueing and early emphasis on functional rehabilitation. CONCLUSIONS: Patients with visual neglect managed on a stroke unit have similar destination of discharge despite lower Barthel Index scores compared with patients of equal stroke severity who do not have this deficit. Spatiomotor cueing and early emphasis on function can improve outcome and reduce resource use in these patients. PMID- 9227690 TI - Prognostic value and determinants of first-day mean arterial pressure in spontaneous supratentorial intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: The onset of spontaneous intracerebral hemorrhage (ICH) is often accompanied by transient blood pressure (BP) elevation. The prognostic value and the determinants of this BP reaction have not entirely been solved, and the present study was focused on these questions. METHODS: From 1985 to 1991 in Central Finland (population, 246,000), a total of 425 patients had first-ever ICH verified by CT or necropsy. The hematoma was supratentorial in 337 patients. Of the 306 patients with supratentorial ICH who had CT, 282 had the BP measured at least once within 24 hours of onset, and they formed the study population. The case notes and CT films were reviewed, and mean arterial pressure (MAP) was calculated from the highest BP reading. RESULTS: The fatality rate was high; 43% of the patients were dead within 28 days of onset. Six independent predictors of the 28-day survival were identified by multiple logistic regression; these predictors were consciousness on admission, first-day MAP, subarachnoid spread of the bleed, lateral shift of hemispheral midline structures, admission blood glucose, and vomiting. The MAPs varied between 66.7 and 203.3 mm Hg, and the cutoff points of the MAP quartiles were 118, 132, and 145 mm Hg. Patients in the first three MAP quartiles had relatively fair outcome, with 71%, 65%, and 60%, respectively, alive 28 days after onset. This was in sharp contrast to the fourth quartile, with only 33% surviving the first 28 days (log-rank, P < .0001 to P = .0010). Patients unconscious/ comatose on admission had significantly higher MAPs than did those who were alert or somnolent/disoriented (ANOVA, P = .0079). However, at all levels of consciousness, the 28-day fatality rate increased from the first to the fourth MAP quartile: 69% in the alert, 186% in the somnolent/disoriented, and 45% in the unconscious/comatose patients. Stepwise multiple regression analysis gave four independent predictors of the first-day MAP: hypertension, age (in an inverse fashion), admission blood glucose, and hematoma volume. CONCLUSIONS: The most important predictor of the 28-day survival was the level of consciousness on admission, followed by first-day MAP. Hypertension was the most important predictor of the first-day MAP, followed by age, which had an inverse effect on the MAP level. At all levels of consciousness, high first-day MAP (especially if > 145 mm Hg) worsened the 28-day survival rate. PMID- 9227689 TI - Polymorphisms of the human platelet antigens HPA-1, HPA-2, HPA-3, and HPA-5 on the platelet receptors for fibrinogen (GPIIb/IIIa), von Willebrand factor (GPIb/IX), and collagen (GPIa/IIa) are not correlated with an increased risk for stroke. AB - BACKGROUND AND PURPOSE: A recent study has described a high incidence of the human platelet antigen (HPA)-1b alloantigen in patients with myocardial infarction. We investigated the distribution of gene polymorphisms of platelet glycoproteins (GPs) in patients with cerebrovascular disease (CVD) and stroke. The polymorphic systems we have studied are HPA-1 and HPA-3 on the fibrinogen receptor (GPIIb/IIIa), HPA-2 on the von Willebrand factor receptor (GPIb/IX), and HPA-5 on one of the platelet collagen receptors (GPIa/IIa). METHODS: DNA was isolated from peripheral blood collected from 218 consecutive stroke patients, 165 neurological inpatients without signs of CVD, and 321 healthy blood donors. The genotypes of HPA-1, HPA-2, HPA-3, and HPA-5 were determined by sequence specific primer polymerase chain reactions. RESULTS: The calculated allele frequencies were as follows: for CVD patients, HPA-1a/b 0.81/0.19, HPA-2a/b 0.91/0.09, HPA-3a/b 0.61/0.39, and HPA-5a/b 0.92/0.08; for inpatient HPA-1a/b 0.83/0.17, HPA-2a/b 0.91/0.09, HPA-3a/b 0.62/0.3 and HPA-5a/b 0.93/0.07; and for blood donors, HPA-1a 0.85/0.15, HPA-2a/b 0.94/0.06, HPA-3a/b 0.60/0.40, and HPA 5a/b 0.92/0.08. There were no statistically significant difference for the analyzed HPA polymorphism frequencies either between the CVD patients and the non CVD inpatients or the CVD patients and blood donors. However, the HPA-1b genotype was slightly more frequent in patients (CVD and non-CVD) than in the healthy blood donors. CONCLUSIONS: Our results indicate that the HPA-1, HPA-2, HPA-3, and HPA-5 polymorphisms are not associated with an increased risk for stroke. PMID- 9227691 TI - Blood pressure changes in acute cerebral infarction and hemorrhage. AB - BACKGROUND AND PURPOSE: We sought to investigate the changes in blood pressure (BP) that occur after hospitalization of patients with different types of acute stroke. METHODS: Twenty-four-hour ambulatory BP monitoring was performed on days 1 and 7 after admission to the hospital in 72 patients with acute stroke (44 thromboembolic strokes, 18 lacunar infarcts, and 10 intracerebral hemorrhages) and in 22 control patients. Stroke was categorized clinically into the above stroke subtypes with radiological confirmation. The controls were patients admitted with a range of acute medical problems other than stroke who were not severely ill or in significant pain. Left ventricular hypertrophy was assessed with echocardiography. Multiple linear regression was used to determine the effect of stroke category on BP after adjustment for the effects of potential confounders. RESULTS: Patients with thromboembolic and lacunar strokes had significantly higher systolic BP (SBP) on day 1 than control subjects (mean, 8.6% and 13.2%, respectively). Diastolic BP (DBP) was also significantly higher for patients with thromboembolic and lacunar strokes on day 1 (mean, 11.7% and 14.6%, respectively). Patients with intracerebral hemorrhage had SBP 9.7% and DBP 6.3% higher than control subjects on day 1, but the results did not achieve statistical significance. By day 7 there was no significant difference in SBP or DBP between the stroke subgroups and control subjects. CONCLUSIONS: BP is elevated after stroke but resolves spontaneously after 7 days. This transient elevation in BP does not appear to result solely from the stress of hospitalization. PMID- 9227692 TI - Spontaneous intracranial hemorrhage: which patients need diagnostic cerebral angiography? A prospective study of 206 cases and review of the literature. AB - BACKGROUND AND PURPOSE: In spontaneous intracerebral hemorrhage (ICH), the site, age of the patients, and preexisting hypertension are important factors in determining the possibility of finding an underlying vascular abnormality by cerebral angiography. To what extent these three factors affect the indication for angiography remains controversial. A prospective study was carried out to correlate the angiographic findings with these three factors. METHODS: Two hundred six consecutive spontaneous ICH cases with an age range from 5 to 79 years (median, 45) were investigated with CT and cerebral angiography over a 3 year period (April 1993 through March 1996). Exclusion criteria were (1) poor surgical risk or severely neurologically disabled patients, (2) refusal of angiography, (3) patients in whom severe coagulopathy accounted for the hemorrhage, (4) bleeding into tumor, or (5) subarachnoid hemorrhage-predominant cases. RESULTS: Angiographic yield (the frequency of positive angiography in a defined patient group) was significantly higher in patients (1) at or below the median age of 45 than those above (53/105, 50%, versus 18/101, 18%; P < .001) and (2) without preexisting hypertension than those with (64/145, 44%, versus 5/58, 9%; P < .001). The correlation of age and preexisting hypertension to angiographic yield was independent (logistic regression coefficients -0.056 and 1.59 and SE 0.12 and 0.515, respectively, both P < .001). In patients of the younger age group without preexisting hypertension, angiographic yield was 48% in putaminal, thalamic, or posterior fossa ICH and 65% in lobar ICH. In the older hypertensive patients, the yields were 0% and 10%, respectively. However, in patients with isolated intraventricular hemorrhage, most were normotensive and the yield was high in both age groups (67% versus 63%). CONCLUSION: Diagnostic cerebral angiography should be considered for all spontaneous ICH patients except those over 45 years old with preexisting hypertension in thalamic, putaminal, or posterior fossa hemorrhage. PMID- 9227693 TI - Interactive effects of age and hypertension on volumes of brain structures. AB - BACKGROUND AND PURPOSE: Advanced age and hypertension have each been associated with changes in brain morphology and cognitive function. To investigate the interaction of age and hypertension with structural brain changes and neuropsychological performance in otherwise healthy patients with essential hypertension, we compared young-old (ages 56 to 69 years) and old-old (ages 70 to 84 years) hypertensive patients (n = 27) with 20 age-matched normotensive healthy control subjects, using quantitative volumetric MRI and a battery of neuropsychological tests. METHODS: Quantitative regions of interest and segmentation analyses were applied to MRI scans of brain to measure volumes of different brain structures and of cerebrospinal fluid (CSF). Severity of white matter hyperintensities (WMHs) was qualitatively rated in the MRI scans. A battery of neuropsychological tests was administered to each subject. RESULTS: The combined hypertensive group (young-old and old-old) had smaller volumes of thalamic nuclei and larger volumes of CSF in the cerebellum and temporal lobes and showed poorer performance in memory and language tests than did the control subjects. Main effects for age were significant in multiple brain regions of interest. The old-old hypertensive patients and age-matched control subjects demonstrated volume reductions in brain structures and increases in ventricular and peripheral CSF volumes compared with the younger subjects. There was a significant group x age-group interaction in temporal and occipital CSF, not related to WMH, with the old-old hypertensive patients having significantly larger CSF volumes in these regions than the young-old hypertensives and both healthy control groups. CONCLUSIONS: Hypertension exacerbates the morphological changes accompanying advanced age. Temporal and occipital regions appear most vulnerable to brain atrophy due to the interactive effects of age and hypertension. PMID- 9227694 TI - Diagnosis of cerebral amyloid angiopathy. Sensitivity and specificity of cortical biopsy. AB - BACKGROUND AND PURPOSE: Examination of cortical tissue obtained surgically is an important tool for diagnosis of cerebral amyloid angiopathy (CAA) during life. Analysis of a single sample of cortical tissue, however, might lead to conclusions that are either falsely positive (because of the high frequency of CAA in the healthy elderly) or falsely negative (because of the patchy distribution of CAA pathology). We therefore attempted to estimate the sensitivity and specificity of cortical biopsy for diagnosis of CAA as the cause of intracerebral hemorrhage. METHODS: To simulate biopsy in CAA, we took biopsy sized cortical samples from postmortem brains with known extents of CAA: either CAA-related hemorrhage or mild to severe CAA without hemorrhage. Samples were stained with the use of methods routinely available in surgical pathology laboratories and blindly examined for vascular amyloid and amyloid-related vasculopathic changes. RESULTS: The presence of vascular amyloid was a sensitive marker for CAA-related hemorrhage, occurring in all 28 specimens from brains with hemorrhage. Conversely, the appearance of fibrinoid necrosis in amyloid-laden vessels was relatively specific for CAA-related hemorrhage. This finding occurred in 13 of the 28 specimens (46%) from brains with hemorrhage but in none of 27 sections from brains with mild CAA and in only 4 of 42 specimens with moderate to severe CAA without hemorrhage. CONCLUSIONS: These data help to define criteria for the diagnosis of CAA-related hemorrhage from surgical specimens. PMID- 9227695 TI - Alterations in glia and axons in the brains of Binswanger's disease patients. AB - BACKGROUND AND PURPOSE: Although increasing attention is being paid to Binswanger's disease, a form of vascular dementia characterized by diffuse white matter lesions, only limited information is available on the pathological changes that occur in the glia and axons in the white matter. We therefore investigated the brains of patients with Binswanger's disease to gain further insight into its pathophysiology. METHODS: Autopsied brains from patients with Binswanger's disease (group 3; n = 17) were compared with those of nonneurological controls (group 1; n = 5) and controls with large cortical infarcts but without significant white matter lesions (group 2; n = 5). Glial fibrillary acidic protein (GFAP) was used as an immunohistochemical marker for astroglia, leukocyte common antigen (LCA) was used as a marker for microglia, and HLA-DR was used as a marker for activated microglia. Axonal damage was assessed by the accumulation of proteins, which are transported by fast axonal flow, amyloid protein precursor (APP), synaptophysin, and chromogranin A. RESULTS: Although there was no difference in numerical density of GFAP-immunoreactive astroglia in each group, regressive astroglia were observed in 7 of 17 patients with Binswanger's disease. LCA-immunoreactive microglia were 1.7 times more numerous in Binswanger's disease than in group 1 (P < .05). HLA-DR-immunoreactive-activated microglia were 3.4 times and 2.1 times more numerous in Binswanger's disease as compared with group 1 (P < .01) and group 2 (P < .05), respectively. There was frequent perivascular lymphocyte cuffing, and clusters of macrophages with a decreased number of oligodendroglia were observed in the rarefied white matter. The grading scores for the number of axons immunoreactive for either APP, synaptophysin, or chromogranin A were significantly higher in Binswanger's disease than in group 1 or 2. CONCLUSIONS: The pathological alterations in Binswanger's diseased brains include regressive changes in the astroglia and activation of the microglia with a decrease in the oligodendroglia, which were associated with the degradation of both myelin and axonal components. These results indicate that an inflammatory reaction and compromised axonal transport, mediated by chronic ischemia, may play an important role in the pathophysiology of Binswanger's disease. PMID- 9227696 TI - CP-0597, a selective bradykinin B2 receptor antagonist, inhibits brain injury in a rat model of reversible middle cerebral artery occlusion. AB - BACKGROUND AND PURPOSE: Recent studies demonstrated a significant neuroprotective action of the selective peptide-based bradykinin B2 receptor antagonist CP-0597 after permanent middle cerebral artery (MCA) occlusion (MCAO) in the rat. We therefore evaluated the efficacy of this compound after reversible MCAO in the rat. METHODS: Male Wistar rats underwent reversible MCAO by insertion of a nylon monofilament to the origin of the MCA. After 1 hour the filament was retracted and the ischemic tissue reperfused. Immediately after MCAO, primed miniosmotic pumps containing either vehicle or CP-0597 (300 ng/kg per minute) were implanted into the subcutaneous space (n = 14 per group). Twenty-four hours after surgery, animals were killed and brains fixed, and 4-micron sections were taken from five sequential tissue blocks labeled A through E and stained with hematoxylin and eosin. Clinical evaluation of rats was performed by neurological scoring and change in body weight. RESULTS: Treatment with CP-0597 significantly reduced percent increase in hemisphere size of the ischemic hemisphere in all brain sections (C section: vehicle, 40.6 +/- 4.3% versus CP-0597, 20.8 +/- 5.3%; P < 0.001), total infarct volume (vehicle, 206.5 +/- 7.7 mm3 versus CP-0597, 94.0 +/- 19.2 mm3; P < .001), cortical infarct volume (vehicle, 145.5 +/- 4.5 mm3 versus CP-0597, 64.0 +/- 15.1 mm3; P < .001), subcortical infarct volume (vehicle, 55.8 +/- 4.1 mm3 versus CP-0597, 27.5 +/- 4.5 mm3; P < .001), and the number of necrotic neurons (vehicle 42.9 +/- 3.8 versus CP-0597, 23.6 +/- 4.7 per field; P < .01). Neurological score (vehicle, 2.78 +/- 0.36 versus CP-0597, 6.29 +/- 0.87 P < .01) and change in body weight (vehicle, -28.7 +/- 2.0 g versus CP-0597, 18.2 +/- 2.8 g; P < .01) were also significantly improved. CONCLUSIONS: The present data demonstrate the significant overall efficacy profile of CP-0597 in a rat model of reversible MCAO and provide strong rationale for the use of such bradykinin B2 receptor antagonist in the treatment of stroke. PMID- 9227697 TI - Multiple-dose mannitol reduces brain water content in a rat model of cortical infarction. AB - BACKGROUND AND PURPOSE: Repeated use of mannitol in the setting of ischemic infarction is a controversial and poorly defined therapeutic intervention. The purpose of this study was to examine the effects of repeated mannitol infusions on brain water content and tissue pressure in a well-defined rat model of focal ischemic stroke. METHODS: Mannitol infusions (0.5, 1.5, or 2.5 g/kg) were given by intravenous bolus 4 or 24 hours after 90-minute transient cortical ischemia in the territory of the right middle cerebral artery in rats and every 4 hours thereafter for a total of 24 hours. Fluid replacement was limited to 0.5 mL i.v. isotonic saline administered immediately after each mannitol dose. Control rats received 0.5 mL i.v. saline at the same intervals and were otherwise under ad libitum conditions. Water contents (percent H2O) of whole hemispheres and of cortical biopsies were measured with the wet-dry method, and blood samples were analyzed for plasma osmolality and chemistries. In a subgroup of rats, tissue pressure was also measured within the hemisphere ipsilateral to the infarct. RESULTS: Repeated mannitol infusions resulted in a dose-dependent increase in plasma osmolality and a dose-dependent decrease in the percent H2O of the ischemic middle cerebral artery cortex and ipsilateral hemisphere. In contrast, percent H2O of the contralateral cortex and hemisphere was significantly decreased only in the groups given the highest dose of mannitol (2.5 g/kg). Mannitol infusions at a dose of 1.5 g/kg begun 24 hours after reperfusion were also associated with a significant reduction of tissue pressure. CONCLUSIONS: In a rat model of ischemic cortical infarction, repeated mannitol infusions resulted primarily in a decrease in the percent H2O of the infarct and ipsilateral hemisphere, as well as decreased tissue pressure. PMID- 9227698 TI - Cushing's 'variant' response (acute hypotension) after subarachnoid hemorrhage. Association with moderate intracranial tensions and subacute cardiovascular collapse. AB - BACKGROUND AND PURPOSE: Hypertension is considered common and appropriate with subarachnoid hemorrhage (SAH), maintaining cerebral perfusion. Hypotension, in contrast, is considered rare and detrimental. This study was designed to assess the frequency of each in both acute and subacute phases of primary SAH. METHODS: SAH was created by arterial rupture in spontaneously breathing rats under urethane anesthesia without craniotomy (n = 32). Arterial pressure and intracranial pressure (ICP) were monitored invasively. RESULTS: After extensive extravasation, the mean ICP rose acutely from 8 +/- 1 to 53 +/- 4 mm Hg over 2.4 +/- 0.3 minutes. Acute pressor changes occurred transiently in 71%. The most common acute response was hypotension (63%). Hypertension, in contrast, was rare (6%); the remainder was invariant (29%). Hypertension was associated with significantly lower maximum ICP values (39 +/- 4 versus 69 +/- 4 mm Hg, P < .001) with a negative correlation between hypotension and delta ICP (r = -.7, P < .01). Distinct and independent of acute responses, hypotension also occurred subacutely as a cardiovascular collapse (38%). CONCLUSIONS: In contrast to popular belief, the most common acute response with SAH is hypotension; hypertension is rare. This, in fact, is in full agreement with Cushing: hypertension was seen only with gradual delta ICPs. In contrast, a "variant" to the classic response (hypotension) occurred with sudden delta ICPs. In the present study, hypotension stanched SAH at lower maximum ICP values, and thus with less cerebral compression. Despite this, cardiovascular collapse developed in a large proportion irrespective of acute change. Such collapse without prior hypertension (94%) implies a nonadrenergic etiology. PMID- 9227699 TI - A near-infrared spectroscopic study of cerebral ischemia and ischemic tolerance in gerbils. AB - BACKGROUND AND PURPOSE: To explore the physiological mechanism of ischemic tolerance, we studied intracerebral oxygenation states noninvasively using near infrared spectroscopy after bilateral common carotid artery occlusion (BCO) in gerbils with and without ischemic pretreatment. METHODS: Under ether anesthesia, gerbils with sham operation (S group, n = 8) and those with pretreatment consisting of BCO for 2 minutes, twice at 3 days and 2 days earlier (T group, n = 8), were again subjected to BCO for 5 minutes. Changes in oxyhemoglobin (HbO2), deoxyhemoglobin (Hb), and total hemoglobin (HbT) as well as reduction in cytochrome oxidase (cyt.aa3) were calculated from the absorbance changes of the light transmitted through the brain. Seven days after the ischemic study, immunohistochemical examination was performed with an antiserum against microtubule-associated proteins. RESULTS: In both groups, the increase of Hb and decrease of HbO2 and HbT proceeded rapidly after BCO, and the maximal deoxygenation of hemoglobin occurred within 2.5 minutes. Reduction of cyt.aa3 also ensued rapidly and reached the maximal reduction within 3 minutes in both groups. In the T group, however, both deoxygenation of hemoglobin and reduction of cyt.aa3 progressed more slowly than in the S group. The time (seconds) necessary for a maximal change for cyt.aa3 was significantly longer in the T group (203.8 +/- 34.0 [mean +/- SD]; P < .01) than in the S group (68.0 +/- 14.7). The time necessary for a half-maximal change was also significantly longer in the T group than in the S group for both Hb (22.0 +/- 7.5 and 13.5 +/- 4.0, respectively; P < .05) and cyt.aa3 (23.9 +/- 5.7 and 11.6 +/- 4.3; P < .01). After recirculation for 7 days, all gerbils in the S group were found to have neuronal death in the hippocampus, while those in the T group did not. CONCLUSIONS: The present study indicated that mild ischemic stress can induce improvement in oxygen metabolism during subsequent ischemia, which might be causally related to the phenomenon known as "ischemic tolerance," in which a protective effect toward ischemic/postischemic injury is induced by earlier mild ischemic pretreatment. PMID- 9227700 TI - Cerebral blood flow in single and multiple lacunar infarctions. AB - BACKGROUND AND PURPOSE: Single and multiple lacunar infarctions may have some difference in underlying diseases and cerebral blood flows. To determine the difference, we investigated underlying diseases and cerebral blood flows in single and multiple lacunar infarctions. METHODS: Fifteen cases of lacunar infarction, 10 cases of multiple lacunar infarctions, and 16 control subjects were studied. Regional cerebral blood flow was measured within 14 days after stroke onset with the stable xenon CT method. RESULTS: The rate of association of diabetes mellitus was higher in the multiple lacunar infarctions group than in the single lacunar infarction group. The blood flow in the cerebral cortex was significantly lower in the multiple lacunar infarctions group than in the single lacunar infarction group. The blood flow change by acetazolamide in the cerebral cortex was significantly lower in the multiple lacunar infarctions group than in the single lacunar infarction group. CONCLUSIONS: There is some difference in underlying diseases and cerebral blood flows between single and multiple lacunar infarctions. PMID- 9227701 TI - 99mTc-HMPAO brain SPECT imaging in a case of repeated syncopal episodes associated with smoking. AB - BACKGROUND: We report here a rare case of repeated syncopal episodes associated with smoking and findings of 99mTc-hexamethylpropyleneamine oxime (HMPAO) brain single-photon emission CT (SPECT) imaging. CASE DESCRIPTION: A 77-year-old man had four syncopal episodes during a half-month period. All four occurred when he stood up and walked immediately after smoking a cigarette, and syncope did not occur after cessation of smoking. Although upright testing revealed orthostatic hypotension, the patient did not complain of fainting on standing alone. Compared with brain SPECT in the supine position, perfusion was decreased in the posterior circulation structures after the subject smoked a cigarette or chewed nicotine gum. CONCLUSIONS: The combination of cerebral vasoconstriction due to smoking and orthostatic hypotension probably decreased cerebral blood flow in this patient, resulting in syncope. PMID- 9227702 TI - Severe pathological crying after left anterior choroidal artery infarct. Reversibility with paroxetine treatment. AB - BACKGROUND: There is increasing evidence that serotonergic neurotransmission may be damaged in poststroke pathological crying. A correlation between the clinical severity of pathological crying and the size of stroke-induced serotonergic pathway lesions is commonly accepted. We present a case of severe pathological crying after a limited left anterior choroidal artery territory infarction. CASE DESCRIPTION: A right-handed 55-year-old man who was a heavy smoker was admitted to the hospital after a right hemiplegia of sudden onset. Clinical examination revealed a right global hemiplegia including the face and a right hemihypoesthesia. Cerebral CT scan and MRI showed an infarct in the retrolenticular part of the posterior limb of the left internal capsule extending upward into the posterior paraventricular corona radiata region. Transesophageal echocardiography revealed an atrial septal aneurysm of 15-mm excursion without associated patent foramen ovale. From the first day of admission, the patient exhibited very frequent and intense fits of pathological crying. Their persistence led to initiation of treatment with the selective serotonin reuptake inhibitor paroxetine on day 30. Complete and immediate resolution of pathological crying occurred 24 hours after onset of therapy. Follow-up examination at day 90 confirmed the absence of relapse of pathological crying. CONCLUSIONS: We conclude that poststroke pathological crying in our patient may have been due to unilateral disruption of the capsular ascending projections of the serotonergic brain stem raphe nuclei. A small left-sided capsular lesion may have led to severe pathological crying. This disabling condition may be reversible with selective serotonin reuptake inhibitor therapy. PMID- 9227703 TI - Serum lipids in acute stroke and a year later. PMID- 9227704 TI - Effects of a novel inhibitor of guanylyl cyclase on dilator responses of mouse cerebral arterioles. PMID- 9227705 TI - Practice guidelines for the use of imaging in transient ischemic attacks and acute stroke. A report of the Stroke Council, American Heart Association. PMID- 9227706 TI - American Heart Association Prevention Conference. IV: Prevention and Rehabilitation of Stroke. Keynote Address: properly designed trials and identification of risk factors. PMID- 9227707 TI - American Heart Association Prevention Conference. IV. Prevention and Rehabilitation of Stroke. Etiology of stroke. PMID- 9227708 TI - American Heart Association Prevention Conference. IV. Prevention and Rehabilitation of Stroke. Risk factors. PMID- 9227709 TI - American Heart Association Prevention Conference. IV. Prevention and Rehabilitation of Stroke. Acute interventions. PMID- 9227710 TI - American Heart Association Prevention Conference. IV. Prevention and Rehabilitation of Stroke. Rehabilitation. PMID- 9227711 TI - Matrix metalloproteinases: a role in emphysema? PMID- 9227712 TI - The aetiology of mesothelioma: are risk factors other than asbestos exposure important? PMID- 9227714 TI - Elevated levels of matrix metalloproteinases in bronchoalveolar lavage fluid of emphysematous patients. AB - BACKGROUND: Matrix degradation in emphysema has long been attributed to the action of neutrophil elastase (NE). More recently a role for other proteases, particularly the matrix metalloproteinases (MMPs), in the pathogenesis of this disease has been proposed. To date, however, the presence of MMPs in the lungs of patients with emphysema has not been demonstrated. METHODS: Samples of bronchoalveolar lavage (BAL) fluid from 10 patients with emphysema and from control subjects matched for sex and current smoking status were assessed for collagenase, gelatinase, and NE activity. Pulmonary function tests and computed tomographic (CT) scans were carried out on all study subjects. RESULTS: Collagenase activity was detected in BAL fluid samples from all emphysematous patients but in only one smoking control (p < 0.001). Gelatinase B was present in six patients and in two smoking controls (p < 0.03). The concomitant presence of gelatinase B in complex with lipocalin (NGAL) in the gelatinase positive samples suggests that the neutrophil is a significant source of the gelatinase B observed. NE was detected in six of the 10 patients with emphysema and in two smoking controls (p < 0.01), indicating that collagenase was more useful in discriminating between disease and control groups than either NE or gelatinase B. No relationship was observed between any of the enzymes measured and pulmonary function or CT density score. CONCLUSIONS: This study demonstrates, for the first time, the presence of increased levels of matrix metalloproteinases in the lungs of patients with emphysema and suggests that, in BAL fluid, collagenase activity may be a better indicator of the presence of emphysema than elastase. PMID- 9227713 TI - The use of induced sputum to investigate airway inflammation. PMID- 9227715 TI - Malignant mesothelioma in south east England: clinicopathological experience of 272 cases. AB - BACKGROUND: Malignant mesothelioma is a rare pleural tumour associated with asbestos exposure. The proportion of malignant mesothelioma unrelated to asbestos exposure, and any differentiating features between exposed and unexposed cases, are not well described. This study describes occupational, clinical, and pathological features in a large cohort of cases of malignant mesothelioma from south east England. METHODS: All 272 cases from this region were studied, either in life or after death when necropsy examination suggested malignant mesothelioma. Detailed information was gathered regarding the occupational history, clinical course, and mode of death. Necropsies were performed in 98% of cases. Lung tissue was examined histologically to confirm the diagnosis, subtype of tumour, presence or absence of asbestosis and asbestos bodies. RESULTS: Exposure to asbestos was documented in 87% of cases, while in the remainder, no asbestos exposure was found nor were asbestos bodies seen; 94.5% were pleural, 5.1% peritoneal, and 0.4% pericardial. Right sided tumours were more common than left sided tumours (ratio 1.6:1). Patients usually presented with breathlessness and chest pain, but 33% presented with pleural effusion in the absence of chest pain. The mean (SD) time from first exposure to asbestos to symptoms was 40 (12) years with a median (interquartile range (IQR) survival of 14 (12.5) months. The median (IQR) survival time in sarcomatous, epithelial, and mixed cell type malignant mesothelioma was 9.4 (10) months, 12.5 (18) months, and 11 (14) months, respectively, and was significantly greater in cases detected by chance. Clinical features were similar in asbestos related and non-asbestos related malignant mesothelioma. CONCLUSIONS: In south east England most cases of malignant mesothelioma are associated with asbestos exposure. Clinical features do not differentiate between asbestos related and non-asbestos related disease. PMID- 9227716 TI - Effect of inhaled prostaglandin D2 in normal and atopic subjects, and of pretreatment with leukotriene D4. AB - BACKGROUND: Prostaglandin (PG) D2 is a potent bronchoconstrictor mediator and is found, together with leukotriene (LT) D4, in bronchoalveolar lavage fluid during the early response to allergen challenge in asthmatic subjects. The potency of PGD2 has not been established in normal and atopic non-asthmatic subjects, nor has the contribution of cholinergic mechanisms to PGD2 induced bronchoconstriction in normal subjects. Mediators released simultaneously may interact, so the effect of pre-inhalation of LTD4 on PGD2 responsiveness was investigated. METHODS: Six normal and six atopic non-asthmatic subjects performed histamine and PGD2 challenges on separate occasions. Eight normal subjects performed PGD2 challenges immediately before and 45 minutes after inhalation of 200 micrograms oxitropium bromide or placebo. Bronchial responsiveness to PGD2 was established in six normal subjects immediately after pretreatment with saline or non-bronchoconstricting doses of methacholine or LTD4 (challenge 1), and again at six hours (challenge 2). All studies were performed in a double blind, randomised, crossover fashion. RESULTS: PGD2 was 25-fold and 18-fold more potent as a bronchoconstrictor than histamine in atopic non-asthmatic and normal subjects, respectively. Responsiveness (PC35sGaw) to histamine and PGD2 correlated significantly (r = 0.917, n = 12, p < 0.001). Oxitropium bromide in a dose of 200 micrograms inhibited PGD2 induced bronchoconstriction by 37.5%, although in two of these subjects no inhibition was seen. Pre-inhalation of LTD4 and methacholine shifted the dose-response curve of PGD2 to the left by 4.6-fold and 2.4-fold, respectively. CONCLUSIONS: PGD2 is a potent bronchoconstrictor in normal subjects, which is partly mediated by cholinergic mechanisms in some subjects. No significant interaction was found between LTD4 and PGD2 in six normal subjects. PMID- 9227717 TI - Effect of pranlukast, an oral leukotriene receptor antagonist, on leukotriene D4 (LTD4) challenge in normal volunteers. AB - BACKGROUND: There is increasing evidence to show that leukotrienes are important mediators in asthma. Leukotriene receptor antagonists protect against antigen and exercise challenges in patients with chronic asthma. A study was undertaken to investigate the activity of the leukotriene receptor antagonist pranlukast (SB 205312, ONO-1078) in blocking bronchoconstriction induced by leukotriene D4 (LTD4) inhalation. The selectivity of pranlukast was evaluated using histamine challenge. METHODS: Pranlukast, 450 mg twice daily, was given to eight healthy non-smoking men for five days in a randomised, double blind, placebo controlled, crossover study. The specific airways conductance (sGaw) was measured before and after bronchial provocation with inhaled LTD4 at 3.5 hours after the first dose and at 3.5 and 9.5 hours after the last dose of pranlukast on the morning of day 5. The concentration of LTD4 required to produce a fall in sGaw of 35% (PC35) was calculated. Subjects also underwent a histamine challenge 3.5 hours after a single dose of pranlukast, 450 mg, or placebo. RESULTS: A single dose of pranlukast produced a 10.6 fold increase in PC35sGaw (95% confidence interval (CI) 4.4 to 25.5; p < 0.001) for LTD4 at 3.5 hours after dosing compared with placebo. Three and a half hours after the morning dose of pranlukast on day 5 the PC35sGaw for LTD4 was increased 25.9 fold (95% CI 10.8 to 62.2; p < 0.001) and was still increased sevenfold (95% CI 2.9 to 16.7; p < 0.001) relative to placebo 9.5 hours after administration of the morning dose. No significant differences were noted for the PC35sGaw to histamine for pranlukast compared with placebo. CONCLUSIONS: This study shows that pranlukast is a potent and selective LTD4 receptor antagonist in humans which blocks LTD4 challenge after initial and repeated administration when given twice daily for five days. PMID- 9227718 TI - Pranlukast, a novel leukotriene receptor antagonist: results of the first European, placebo controlled, multicentre clinical study in asthma. AB - BACKGROUND: Leukotriene receptor antagonists have been shown to protect against bronchoconstriction induced by antigens, exercise, and cold air. There are relatively few clinical studies reported in patients with asthma. The present study is the first clinical evaluation of pranlukast (SB 205312, ONO-1078) outside Japan in patients with asthma. METHODS: A randomised, double blind, placebo controlled, parallel group, multicentre four week study of the safety and tolerability of oral pranlukast, 225 or 337.5 mg twice daily, was performed in patients with mild to moderate asthma. Preliminary efficacy data were obtained; the main efficacy variables evaluated were forced expiratory volume in one second (FEV1) and morning domiciliary (home) peak expiratory flow rates (PEFR). Clinic PEFR and daytime and night-time asthma symptom scores were also recorded. RESULTS: Compared with the placebo group the improvement in morning home PEFR was statistically significant at all time points for patients receiving pranlukast 337.5 mg twice daily and at weeks 1 and 2 for those treated with pranlukast in a dose of 225 mg twice daily. Mean morning home PEFR increased by 10.8 to 18.61/min (95% CI 0.2 to 29.3 l/min) in patients treated with pranlukast compared with a slight deterioration in those given placebo. FEV1 significantly increased within one hour after the first dose of pranlukast compared with baseline and this increase was maintained for eight hours. Improvements in trough FEV1-that is, at the end of the dosing interval-were statistically significant for the group treated with pranlukast 225 mg twice daily compared with placebo at week 4. Mean increases in FEV1 ranged from 210 ml to 340 ml (95% CI 60 to 500 ml) at trough in the pranlukast group. Patients treated with pranlukast also showed improvements in summary symptom and night-time asthma scores. Pranlukast was well tolerated, and no drug related changes in haematological and biochemical variables were observed. CONCLUSIONS: Pranlukast, an oral leukotriene receptor antagonist, is well tolerated and is effective for the treatment of asthma. It increased FEV1 within one hour of dosing, improved patient summary symptom and night-time asthma scores, and reduced the use of rescue bronchodilators, thus providing further evidence of a role for leukotrienes in the pathogenesis of asthma. PMID- 9227719 TI - Climate and aeroallergen levels in asthma: a 12 month prospective study. AB - BACKGROUND: There is evidence to suggest that changes in weather and airborne fungal spore and pollen counts may affect asthma symptoms. METHODS: The relationship between climate, airborne fungal spore, and pollen counts and peak expiratory flow rate (PEFR) and asthma symptoms was prospectively investigated in a population of mild to moderate asthmatic subjects in Blenheim, New Zealand. Subjects recorded twice daily PEFR measurements and asthma symptom scores for up to one year. Spore and pollen counts were measured two hourly and meteorological data were measured hourly. Individual, within person, multiple linear regression analyses were conducted, adjusting for auto-correlation. A random effects model was assumed for the individual regression co-efficients and weighted estimates of the mean of these coefficients were obtained by the method of maximum likelihood. RESULTS: One hundred and thirty nine asthmatic patients (60% atopic) aged 17-80 years completed the study. Of the weather variables, only temperature showed a small but consistent association with PEFR. The mean rise in PEFR for an 8.8 degrees C (2 SD) change in temperature was 0.78% (95% CI 0.44% to 1.11%), approximately 3.0 l/min. There was a weak association between days of high basidiospore counts and increased nocturnal wakening and reliever medication use. Pollen counts showed no consistent association with either PEFR or asthma symptoms. CONCLUSIONS: The results of this study suggest that the effects of weather and aeroallergens on PEFR and asthma symptoms in this population are small, and that other causes need to be sought to account for variations in asthma severity and exacerbations. PMID- 9227721 TI - Nasal contribution to exhaled nitric oxide during exhalation against resistance or during breath holding. AB - BACKGROUND: The concentration of nitric oxide (NO) is increased in the exhaled air of patients with inflammation of the airways, suggesting that this may be a useful measurement to monitor inflammation in diseases such as asthma. However, there have been concerns that exhaled NO may be contaminated by the high concentrations of NO derived from the upper airways, and that this may account for differences in reported values of exhaled NO using different techniques. A study was performed, with argon as a tracer, to determine the extent of nasal contamination of exhaled NO using different expiratory manoeuvres. METHODS: Exhaled and nasal NO were measured by a chemiluminescence analyser. Argon (4.8%) was delivered continuously to the nose. Gas was sampled from the posterior oropharynx and argon and carbon dioxide were measured by mass spectrometry at the same time as NO. RESULTS: During a single expiration against a low resistance and during breath holding there was no evidence for nasal contamination, whereas during exhalation without resistance argon concentration in the oropharynx was increased from 0.91% (95% CI 0.84% to 0.98%) in ambient air to 1.28% (0.9% to 2.24%, p < 0.0001) during a single breath or 2.37% (2.29% to 2.51%, p < 0.0001) during tidal breathing. CONCLUSIONS: Collection of exhaled NO in a reservoir during tidal breathing is likely to be contaminated by NO derived from the nose and this may underestimate any increases in NO derived from the lower respiratory tract in inflammatory diseases. However, with slow expiration against a resistance and created back pressure to close the soft palate, there is no contamination of exhaled air which then reflects concentrations of NO in the lower airways. PMID- 9227720 TI - Effects of the long acting beta agonist formoterol on asthma control in asthmatic patients using inhaled corticosteroids. The Netherlands and Canadian Formoterol Study Investigators. AB - BACKGROUND: The long acting beta 2 agonist formoterol has proved to be an effective bronchodilator with a prolonged action of 12-14 hours. However, the precise role of formoterol in the maintenance treatment of asthma is still under debate. A study was performed to investigate the efficacy and safety of treatment with formoterol for six months in subjects with asthma. METHODS: In a multicentre double blind, placebo controlled, parallel group study 239 subjects with mild to moderate asthma were randomly assigned to treatment with either inhaled formoterol 24 micrograms twice daily (n = 125) or placebo (n = 114) during eight months. The study consisted of a four week run in period, a 24 week treatment period, and a four week washout period. All subjects were using regular inhaled corticosteroids (100-3200 micrograms daily) but were still needing at least five inhalations of short acting beta 2 agonist per week for symptom relief. The study was performed in 10 outpatient clinics in Canada, and five outpatient clinics and one coordinating centre for 44 Dutch general practitioners in The Netherlands. Twice daily self-reported peak expiratory flow (PEF) measurements, symptom scores, and rescue beta 2 agonist use during the last 28 treatment days compared with baseline values were used as main outcome measures. Spirometric values were measured at entry, at the start of treatment, after four, 12 and 24 weeks of treatment, and after four weeks washout. RESULTS: One hundred and twenty five subjects received formoterol 24 micrograms twice daily via Turbohaler and 114 received placebo. Baseline FEV1 was 67.1% predicted and mean bronchodilator reversibility was 26%. The mean total asthma symptom score was 3.6 (maximum possible 21). A significant decrease in symptoms in favour of formoterol (difference from placebo -0.64, 95% CI -0.04 to -1.23, p = 0.04) was observed. Compared with placebo, morning PEF increased (difference from placebo 28 l/min, 95% CI 18.3 to 37.7, p = 0.0001) and the use of short acting beta 1 agonists decreased (daytime difference from placebo -1.1 inhalation, 95% CI -1.4 to -0.7, p = 0.0001) in the formoterol group. PEF returned to baseline following discontinuation of formoterol, as did asthma symptom scores. Thirty three patients treated with formoterol and 32 treated with placebo required treatment with prednisolone during the study (58 and 55 courses, respectively). CONCLUSIONS: Adding formoterol 24 micrograms twice daily by Turbohaler to inhaled corticosteroids was effective in improving symptom scores and morning PEF, and decreasing the use of rescue beta 2 agonists. There was no apparent loss of asthma control during 24 weeks of treatment with formoterol. PMID- 9227722 TI - Changes in ventilatory mechanics and diaphragmatic function after lung volume reduction surgery in patients with COPD. AB - BACKGROUND: Lung volume reduction (LVR) has recently been used to treat severe emphysema. About 25% of the volume of each lung is removed with this method. Little is known about the mechanism of functional improvement so a study was undertaken to investigate the changes in ventilatory mechanics and diaphragmatic function in eight patients after LVR. METHODS: Measurements of work of breathing (WOB), intrinsic positive end expiratory pressure (PEEPi), dynamic compliance (Cdyn), and arterial carbon dioxide tension (PaCO2) were performed on the day before surgery and daily for seven days after surgery, as well as one, three, and six months after surgery. All measurements were performed on spontaneously breathing patients, simultaneously assessing oesophageal pressure via an oesophageal balloon catheter and air flow via a tightly adjusted mask. Diaphragmatic function was evaluated by measuring oesophageal and transdiaphragmatic pressure (Pdi) preoperatively and at one, three, and six months postoperatively. RESULTS: Mean forced expiratory volume in one second (FEV1) was 23 (3.6)% predicted, and all patients were oxygen dependent before the operation. One day after LVR the mean decrease in WOB was 0.93 (95% confidence interval (CI) 0.46 to 1.40) joule/l, the mean decrease in PEEPi was 0.61 (95% CI 0.35 to 0.87) kPa, and the mean increase in Cdyn was 182.5 (95% CI 80.0 to 284.2) ml/kPa. Similar changes were found seven days and six months after surgery. PaCO2 was higher on the day after the operation but was significantly reduced six months later. Pdi was increased three and six months after surgery. CONCLUSIONS: Ventilatory mechanics improved immediately after LVR, probably by decompression of lung tissue and relief of thoracic distension. An improvement in diaphragmatic function three and six months postoperatively also contributes to improved respiratory function after LVR. PMID- 9227723 TI - Bronchial hyperresponsiveness in lung transplant recipients: lack of correlation with airway inflammation. AB - BACKGROUND: Bronchial hyperresponsiveness (BHR) to methacholine has been reported to occur in most lung transplant recipients. BHR to physical stimuli such as exercise and non-isotonic aerosols has not been as extensively studied in this subject population. This report aims to assess the presence and degree of BHR to methacholine and hypertonic saline in stable lung transplant recipients and to relate it to the presence of airway inflammation. METHODS: Ten patients undergoing bilateral sequential lung transplantation and six heart-lung transplant recipients, all with stable lung function, were recruited 66-1167 days following transplantation. Subjects underwent a methacholine challenge and bronchoscopy for sampling of bronchoalveolar lavage fluid, transbronchial and endobronchial biopsy tissues. Hypertonic saline challenge was performed six days later. RESULTS: Nine of the 16 transplant recipients had positive methacholine challenges (geometric mean PD20 0.18 mg, interquartile range 0.058-0.509) and three of these subjects also had positive hypertonic saline challenges (PD15 = 2.3, 33.0, and 51.5 ml). No clear relationship was found between BHR to either methacholine or hypertonic saline and levels of mast cells, eosinophils or lymphocytes in samples of biopsy tissue or lavage fluid. CONCLUSIONS: Most of the lung transplant recipients studied were responsive to methacholine and unresponsive to hypertonic saline. BHR was not clearly related to airway inflammation, suggesting an alternative mechanism for BHR following lung transplantation from that usually assumed in asthma. PMID- 9227724 TI - Pulmonary oxidative stress response in young children with cystic fibrosis. AB - BACKGROUND: It has been suggested that oxidative stress contributes to lung injury in cystic fibrosis. There is, however, no direct evidence of increased pulmonary oxidative stress in cystic fibrosis nor of the effects of inflammation on the major pulmonary antioxidant, glutathione. A study was undertaken to measure these parameters in infants and young children in the presence or absence of pulmonary inflammation. METHODS: Thirty two infants and young children with cystic fibrosis of mean (SD) age 21.4 (15.3) months (range 2-54) and seven non cystic fibrosis control subjects of mean (SD) age 21.0 (21.2) months (range 2-54) were studied using bronchoalveolar lavage (BAL). On the basis of the BAL findings the cystic fibrosis group was divided into those with (CF-I) and those without pulmonary inflammation (CF-NI). Levels of lipid hydroperoxide, total glutathione, and gamma-glutamyl transpeptidase (gamma-GT) were then measured in the BAL fluid. RESULTS: The concentrations of lipid hydroperoxide and gamma-GT in the epithelial lining fluid were significantly increased in the CF-I group compared with the control and CF-NI groups, each of which had similar values for these parameters (ratio of geometric means for CF-I group versus control for lipid hydroperoxide 5.4 (95% confidence interval (CI) 1.8 to 15.8) and for gamma-GT 5.2 (95% CI 1.4 to 19.4)). The glutathione concentration tended to be lower in the CF-I subjects but the difference did not reach statistical significance. CONCLUSIONS: These results demonstrate that the airways in patients with cystic fibrosis are exposed to increased oxidative stress which appears to be a consequence of pulmonary inflammation rather than part of the primary cystic fibrosis defect. The increase in gamma-GT in the CF-I group suggests a mechanism by which extracellular glutathione could be utilised by airway epithelial cells. PMID- 9227725 TI - Short wave diathermy for small spontaneous pneumothorax. AB - BACKGROUND: The treatment of small spontaneous pneumothorax can involve observation, tube thoracostomy, and surgery. This study evaluated the use of short wave diathermy as a method of accelerating the resolution of small pneumothoraces. METHODS: Twenty two patients with pneumothoraces of less than 30% by volume were randomly allocated to receive short wave diathermy for 25 minutes each day (n = 11) or observation with bed rest (n = 11). Chest radiographs were taken until the pneumothoraces resolved. RESULTS: There were no significant differences in the clinical characteristics between the two groups of patients. However, the mean (SD) rate of absorption was significantly higher with short wave diathermy than with observation (3.44 (0.94)% versus 1.57 (0.53)%, difference = 1.87, 95% confidence interval (CI) 1.19 to 2.55, p < 0.001). The time to complete reexpansion was shorter with short wave diathermy than with observation (6.86 (3.51) days versus 11.64 (3.61) days, difference = -4.78 days, 95% CI -7.95 to -1.61, p < 0.005). No evidence of damage resulting from short wave diathermy was found. CONCLUSIONS: Although further study is necessary, these results indicate that short wave diathermy may be an alternative treatment for patients with small spontaneous pneumothoraces. PMID- 9227726 TI - Prognostic significance of plasma D-dimer levels in patients with lung cancer. AB - BACKGROUND: The peripheral blood concentrations of several proteases of the clotting system have been shown to predict survival in patients with malignancy. A study was undertaken to investigate the independent value of the plasma levels of the D-dimer degradation product of fibrin before treatment for predicting prognosis in patients with lung cancer. METHODS: The study comprised 70 patients with lung cancer (49 non-small cell lung cancer and 21 small cell lung cancer). Plasma levels of D-dimer were measured using an enzyme immunoassay kit. Multivariate statistical analysis was carried out using the Cox's proportional hazards model. RESULTS: The median value of the plasma level of D-dimer differentiated two groups of patients with different outcomes: a group with a D dimer level of < 150 ng/ml (low DD group) and those with D-dimer levels of > or = 150 ng/ml (high DD group). Survival time was significantly better in patients in the low DD group than in those in the high DD group in all patients (hazard ratio for high DD group = 4.7; 95% confidence interval (CI) 1.8 to 11.7). The plasma levels of D-dimer predicted survival independently from the clinical stage of disease, histological type, performance status, and tumour size (hazard ratio = 3.9; 95% CI 1.6 to 9.2). CONCLUSIONS: These results suggest that plasma levels of D-dimer might be useful for predicting the clinical outcome in patients with lung cancer. However, further prospective studies are needed in a larger population to confirm these findings. PMID- 9227727 TI - Attenuation of oxidant/antioxidant imbalance during treatment of exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: An oxidant/antioxidant imbalance is thought to occur in patients with chronic obstructive pulmonary disease (COPD). It has recently been shown that during exacerbations of COPD the antioxidant capacity and protein sulfhydryls of plasma are lower and the levels of products of lipid peroxidation are higher than in age matched healthy subjects. The aims of this study were to confirm these data and to measure the time course of these changes. METHODS: The plasma Trolox equivalent antioxidant capacity (TEAC), protein sulfhydryls, and products of lipid peroxidation were measured throughout the course of treatment in 13 patients who presented with an acute exacerbation of COPD. RESULTS: TEAC values (mmol/l) were low on admission (mean 0.67, 95% confidence interval (CI) 0.32 to 0.88; p < 0.05) and had increased by discharge (0.98, 95% CI 0.88 to 1.2; p < 0.05) but still remained lower than in healthy subjects (1.33, 95% CI 1.11 to 1.65). There was also restoration of plasma protein sulfhydryl levels (mmol/l) from admission (0.32, 95% CI 0.20 to 0.43) to discharge (0.49, 95% CI 0.42 to 0.62, p < 0.001) to levels similar to those in healthy subjects (0.52, 95% CI 0.43 to 0.65). Products of lipid peroxidation, measured as thiobarbituric acid malondialdehyde adducts (mumol/l), were significantly higher (2.08, 95% CI 1.8 to 2.5) than in control subjects (1.3, 95% CI 0.85 to 1.32; p < 0.01) and returned to normal values by the time of discharge (1.2, 95% CI 0.88 to 1.29). CONCLUSIONS: These data confirm the presence of a profound oxidant/antioxidant imbalance in the blood of patients with acute exacerbations of COPD which returns towards normal values during the course of treatment. PMID- 9227728 TI - "Ultimate activation" of eosinophils in vivo: lysis and release of clusters of free eosinophil granules (Cfegs). PMID- 9227730 TI - Spontaneous haemothorax associated with von Recklinghausen's disease: review of occurrence in Japan. AB - The case history is presented of a 61 year old man with von Recklinghausen's disease who developed a spontaneous haemothorax. In spite of being asymptomatic for five days after drainage, he died as a result of fatal sudden re-bleeding. The post mortem examination showed dissection and rupture of the left subclavian artery. Microscopically, disarrangement of smooth muscle and decrease of elastic fibre was observed in the ruptured artery. Haemothorax in patients with von Recklinghausen's disease may require thoracotomy, even if the condition of the patient appears to be stable. PMID- 9227731 TI - Spontaneous pneumothorax in a patient with an azygos lobe. AB - The association between a spontaneous pneumothorax and an azygos lobe is surprisingly rare. A case is reported in which surgical management was difficult; it is suggested that thoracotomy is preferable to video-assisted thoracoscopic surgery in this situation. It is possible that the presence of an azygos lobe might protect against the subsequent development of a spontaneous pneumothorax, and the possible mechanism of this is discussed. PMID- 9227732 TI - Delayed pneumothorax after CT-guided percutaneous fine needle aspiration lung biopsy. AB - Two patients are described who developed pneumothoraces more than 24 hours after computed tomography (CT) guided percutaneous fine needle aspiration lung biopsies. The pneumothoraces required treatment in both cases. Such delayed pneumothorax after lung biopsy is extremely unusual. Patients should be warned of the possible occurrence of this complication and instructed to seek medical help if they develop chest pain or breathlessness. PMID- 9227734 TI - Inhalation drug delivery from seven different spacer devices. PMID- 9227733 TI - Neuromuscular blockade with acute respiratory failure in a patient receiving cibenzoline. AB - Cibenzoline is a class Ic antiarrhythmic agent that can be used to treat supraventricular arrhythmias. A case is reported of cibenzoline overdose in a patient with impaired renal function, leading not only to the usual cardiac and metabolic symptoms (bradycardia and hypoglycaemia), but also to a myastheniform syndrome with acute respiratory failure. Neuromuscular blockade was demonstrated by repetitive supramaximal stimulation of the median nerve, and diaphragmatic involvement was evidenced by applying the same protocol to the phrenic nerve. Muscle strength recovered as serum cibenzoline levels decreased, allowing the patient to be weaned from the ventilator. This observation suggests that cibenzoline, like other antiarrhythmic agents, can be responsible for neuromuscular blockade, and should therefore be used with caution in patients with neuromuscular and respiratory diseases or with impaired renal function. PMID- 9227735 TI - Optimal particle size for aerosols. PMID- 9227736 TI - Dual infection with HIV-1 and HIV-2. PMID- 9227737 TI - Treatment of paraquat poisoning. PMID- 9227738 TI - Growth factors and granulomatous inflammation. PMID- 9227739 TI - Therapeutic roles of animals. PMID- 9227741 TI - Minimizing feline sarcomas. PMID- 9227740 TI - Distinction between "Dr." and "veterinarian". PMID- 9227742 TI - The art of employee involvement and participatory management for food animal veterinarians. PMID- 9227743 TI - What is your diagnosis? Atresia of a portion of the large colon. PMID- 9227744 TI - Theriogenology question of the month. Induction of parturition. PMID- 9227745 TI - Results of the AVMA survey of US pet-owning households on companion animal ownership. PMID- 9227746 TI - Serum biochemical values in sled dogs before and after competing in long-distance races. AB - OBJECTIVE: To measure and compare blood values in sled dogs before and after long distance racing. DESIGN: Prospective study. ANIMALS: 17 adult sled dogs in the 1991 Iditarod Trail Sled Dog Race and 21 in a simulated sled dog race. PROCEDURE: Blood samples were obtained from 17 dogs 7 days before they began and after they finished (finisher group) or were eliminated from (nonfinisher group) the Iditarod Trail Sled Dog Race. Blood samples were also obtained from 21 dogs before and after a simulated sled dog race. RESULTS: In finisher-group dogs, BUN and uric acid (UA) concentrations were increased after racing; nonfinisher-group dogs had significantly lower postrace BUN and UA concentrations. Significant increases in creatine kinase (CK) and aspartate transferase (AST) activities were detected in all dogs after racing, and postrace values were higher in nonfinisher group dogs, compared with finisher-group dogs. Mean alkaline phosphate activities were significantly increased after racing in nonfinisher-group dogs only. In dogs that ran the simulated race, postrace values for serum albumin, total protein, calcium, and potassium concentrations, as well as Hct, hemoglobin concentration, and RBC count, were significantly lower than prerace values. Postrace values for alkaline phosphate, alanine transaminase, AST, lactate dehydrogenase, CK, BUN, and UA were significantly higher than prerace values. CLINICAL IMPLICATIONS: High CK activities are indicative of severe muscle degeneration and, in sled dogs, may represent a degree of muscle breakdown beyond which a dog cannot continue to work. Markedly high CK, and possibly AST, serum activities may be indicators of performance failure in sled dogs competing in long-distance races. PMID- 9227747 TI - Effects of castration on problem behaviors in male dogs with reference to age and duration of behavior. AB - OBJECTIVE: To determine whether 9 problem behaviors in adult male dogs were affected by castration and to examine the influence of age and duration of problem behavior on behavioral effects of castration. DESIGN: Cohort study. ANIMALS: 57 male dogs > 2 years old at the time of castration that had > or = 1 of the targeted problem behaviors. PROCEDURE: Data were collected by telephone contact with owners to identify dogs that had > or = 1 problem behavior before castration and to estimate the improvement (ie, decrease) in the objectionable behaviors after castration. Problem behaviors of interest included urine marking in the house, mounting, roaming, fear of inanimate stimuli, aggression toward human family members, aggression toward unfamiliar people, aggression toward other dogs in the household, aggression toward unfamiliar dogs, and aggression toward human territorial intruders. RESULTS: Effects of castration on fear of inanimate stimuli or aggression toward unfamiliar people were not significant. For urine marking, mounting, and roaming, castration resulted in an improvement of > or = 50% in > or = 60% of dogs and an improvement of > or = 90% in 25 to 40% of dogs. For remaining behaviors, castration resulted in an improvement of > or = 50% in < 35% of dogs. Significant correlations were not found between the percentage of improvement and age of the dog or duration of the problem behavior at the time of castration. CLINICAL IMPLICATIONS: Castration was most effective in altering objectionable urine making, mounting, and roaming. With various types of aggressive behavior, including aggression toward human family members, castration may be effective in decreasing aggression in some dogs, but fewer than a third can be expected to have marked improvement. Age of the dog or duration of the problem behavior does not have value in predicting whether castration will have a beneficial effect. PMID- 9227748 TI - Clinical signs and diagnosis of osteogenesis imperfecta in three dogs. AB - When a young dog is evaluated for multiple fractures with minimal to no accompanying trauma, the primary differential diagnoses are metabolic disease, physical abuse, and osteogenesis imperfecta (OI). Of these, secondary hyperparathyroidism is most common, but if serum concentrations of ionized calcium, phosphorus, vitamin D, and parathormone are within reference ranges, OI must be considered. Osteogenesis imperfecta is a heritable disease characterized by brittle bones. Results of studies using cultured skin fibroblasts indicate that most cases of OI in human beings are caused by a mutation in a type-I collagen gene. Osteogenesis imperfecta was recently identified in 3 dogs. Radiographic findings included multiple fractures in various stages of healing and generalized osteopenia. Cultured fibroblasts from skin biopsy specimens were used to diagnose OI. Structural abnormalities were found in type-I collagen from each dog. This cell culture assay can be used to evaluate dogs with brittle bones. PMID- 9227749 TI - Diabetic ketosis and ketoacidosis in cats: 42 cases (1980-1995). AB - OBJECTIVE: To determine clinical signs, clinicopathologic abnormalities, prevalence of concurrent disease, treatment, complications of treatment, and outcome in cats with diabetic ketosis (DK) or diabetic ketoacidosis (DKA). DESIGN: Retrospective study. ANIMALS: 42 cats with DK or DKA. PROCEDURE: Medical records of diabetic cats with ketonuria were reviewed. RESULTS: In 26 cats, diabetes was newly diagnosed; in 16, diabetes had been diagnosed previously and cats had been treated with insulin (n = 14) or sulfonylurea drugs (2). Common clinical findings were lethargy, anorexia, polyuria, polydipsia, and weight loss. Common laboratory findings were hyperglycemia, hyponatremia, hypochloremia, hypokalemia, hypocalcemia, hypophosphatemia, low total CO2 content, hyperosmolality, high serum alanine transaminase activity, azotemia, glycosuria, and ketonuria. Concurrent disorders were identified in 39 cats and included hepatic lipidosis, cholangiohepatitis, pancreatitis, chronic renal failure, urinary tract infection, and neoplasia. Treatment of DK and DKA included administration of regular crystalline (34 cats), NPH (6), or ultralente (2) insulin, intravenous (38) or subcutaneous (4) administration of fluids, and enterall parenteral or administration of antibiotics (42). Complications during treatment included abnormalities in serum electrolyte concentrations (27 cats), hemolytic anemia (4), hypoglycemia (3), and neurologic abnormalities unrelated to hypoglycemia (2). Eleven cats died or were euthanatized during the initial hospitalization period for treatment of DK or DKA. Azotemia, metabolic acidosis, and hyperosmolality were more severe in cats that died than in cats that survived. Differences in regard to treatment or complications were not apparent between cats that died and cats that survived. The 31 cats that survived were discharged 1 to 16 days (median, 5 days) after initiation of insulin treatment. Diabetic ketosis or ketoacidosis recurred in 13 (42%) of these cats. CLINICAL IMPLICATIONS: A thorough diagnostic evaluation should be performed on cats with DK or DKA to identify concurrent disorders, formulate an appropriate treatment plan, and provide prognostic information to the owner. PMID- 9227750 TI - Field study of the safety, immunogenicity, and efficacy of an inactivated equine rotavirus vaccine. AB - OBJECTIVE: To determine safety, immunogenicity, and efficacy of an inactivated equine rotavirus vaccine. DESIGN: Prospective randomized controlled trial. ANIMALS: 316 pregnant Thoroughbred mares during the first year of the study and 311 during the second year. PROCEDURE: During the first year, mares received 3 doses of vaccine or placebo, IM, at 8, 9, and 10 months of gestation. Serum neutralizing antibody titers were measured before vaccination and 1 and 35 days after foaling. Antibody titers were measured in foals 1, 7, 35, 60, 90, and 120 days after birth. During the second year, mares that had been vaccinated the previous year received a single booster dose of vaccine approximately 1 month prior to parturition. Mares that had received the placebo the previous year and mares new to the study received 3 doses of vaccine or placebo. Serum neutralizing antibody titers were measured in samples taken from mares approximately 1 day after foaling and from foals approximately 1 and 60 days after birth. RESULTS: Adverse reactions were not observed. Antibody titers were significantly increased at the time of foaling and 35 days after foaling in vaccinated, compared with control, mares and for 90 days after birth in foals born to vaccinated, compared with foals born to control, mares. Incidence of rotaviral diarrhea was lower in foals born to vaccinated, compared with foals born to control, mares, but the difference was not significant. CLINICAL IMPLICATIONS: Results suggest that the equine rotavirus vaccine is safe and immunogenic and that reasonable efficacy under field conditions can be expected. PMID- 9227751 TI - Evaluation of itraconazole-dimethyl sulfoxide ointment for treatment of keratomycosis in nine horses. AB - OBJECTIVE: To evaluate the efficacy of itraconazole-dimethyl sulfoxide ointment for treatment of keratomycosis in horses in the northeastern United States. DESIGN: Prospective clinical trial. ANIMALS: 9 horses (10 affected eyes). PROCEDURE: All horses treated for keratomycosis at Cornell University between July 1994 and July 1996 were included in the study. The diagnosis of keratomycosis was confirmed by cytologic examination, and all horses were treated with 0.25 ml of a 1% itraconazole-30% dimethyl sulfoxide petrolatum-based ointment, applied to the affected eye every 4 hours. RESULTS: Topical application of itraconazole-dimethyl sulfoxide ointment (q 4 h) resolved keratomycosis in 8 of 10 eyes; mean duration of treatment was 34.6 days (range, 16 to 53 days). CLINICAL IMPLICATIONS: Results of this study indicate topical administration of itraconazole-dimethyl sulfoxide ointment may provide an additional treatment option for horses with keratomycosis. PMID- 9227752 TI - Ovariectomy of granulosa cell tumors in mares by use of the diagonal paramedian approach: 12 cases (1989-1995). AB - OBJECTIVE: To describe the short- and long-term survival rates in horses undergoing ovariectomy for granulosa cell tumors by use of the diagonal paramedian approach. DESIGN: Retrospective case study. ANIMALS: 12 horses with granulosa cell tumors. PROCEDURE: A diagonal paramedian approach for unilateral ovariectomy was used for removal of each mare's granulosa cell tumor. Information about complications and outcomes was analyzed. RESULTS: Only minimal complications were detected postoperatively when the diagonal paramedian approach was used, regardless of the preferred technique for ovarian pedicle ligation or incisional closure and the use of pre- and postoperative medications. Clinical signs of moderate or severe postoperative abdominal pain were not evident in any of the 12 horses. Short- and long-term survival rates were 100%. CLINICAL IMPLICATIONS: The diagonal paramedian approach was advantageous for ovarian tumor removal, because the ovary was immediately adjacent to the body wall at a portion of the incision site. Size of the ovary was not a limitation, because muscle tissues at the edges of the incision were flexible and easily retractable. All of these factors improved exposure, decreased traction on the ovary, increased our ability to observe the vasculature, and decreased postoperative morbidity, aiding in the removal of granulosa cell tumors in mares. PMID- 9227753 TI - Effects of antimicrobial treatment at the end of lactation on milk yield, somatic cell count, and incidence of clinical mastitis during the subsequent lactation in a dairy herd with a low prevalence of contagious mastitis. AB - OBJECTIVE: To determine whether treating cows with antimicrobials at the end of lactation would lower the incidence of clinical mastitis, improve milk production, and decrease somatic cell count (SCC) in the subsequent lactation. DESIGN: Randomized blind field trial. ANIMALS: 233 Holstein cows from a single herd. All cows were in lactation 2 or greater. PROCEDURE: Cows were randomly assigned to treatment groups. Treated cows were given procaine penicillin G and novobiocin by intramammary infusion. Control cows were not treated. Farm personnel recorded cases of clinical mastitis. Milk yield and SCC were recorded during the subsequent lactation. RESULTS: Treatment did not significantly reduce the incidence of clinical mastitis when data for all cows were grouped or when data were stratified by lactation groups (lactation 2 vs lactation > or = 3) or by last SCC (< or = 500,000 cells/ml vs > 500,000 cells/ml). Somatic cell counts (first, mean of first 5, maximum of first 5) for treated and control cows were similar, and proportions of treated and control cows with SCC > 500,000 cells/ml at least once were not significantly different. Treated cows produced 179 kg (394 lb) more milk during the first 17 weeks of lactation than did control cows. CLINICAL IMPLICATIONS: Treating cows with antimicrobials at the end of lactation increased 17-week milk production during the subsequent lactation and, at current milk prices, was financially preferable to not treating them. PMID- 9227755 TI - Classification of atrial fibrillation. PMID- 9227754 TI - Urinary diagnostic indices in calves. AB - OBJECTIVE: To establish reference values for urinary diagnostic indices in healthy calves from birth to 90 days of age. DESIGN: Prospective field trial. ANIMALS: 12 Holstein heifer calves. PROCEDURE: Urine and serum samples were collected daily for the first 5 days after birth, then weekly until calves were 90 days old. Urine:serum creatinine ratio, urine:serum urea nitrogen ratio, urine:serum osmolality ratio, fractional clearances of sodium and inorganic phosphate, and urine gamma-glutamyltransferase activity were measured. Data were grouped by age of calves at the time of sample collection: 1 to 5 days old (neonatal period), 7 to 27 days old (suckling period), and 28 to 90 days old (weanling period). RESULTS: Mean urine:serum creatinine, urea nitrogen, and osmolality ratios were significantly higher during the weanling period than during the other 2 periods. There were no significant differences in mean fractional clearances of sodium among age periods. CLINICAL IMPLICATIONS: Urinary diagnostic indices calculated for these healthy calves may be used as reference values for early recognition of renal damage or renal failure. PMID- 9227756 TI - Comparison of single-biphasic versus sequential-biphasic shocks on defibrillation threshold in pigs. AB - Current generation implantable cardioverter defibrillators use monophasic, biphasic, or sequential pulse shocks, most of which truncate after a given time, dumping the remaining charge on the capacitor through an internal resistor. We hypothesized that having an additional current pathway, and delivering the majority of the remaining charge on a single capacitor to the two pathways using additional shock phases, would improve defibrillation efficacy. This hypothesis was tested by comparing DFTs using a simulated single capacitor, single-biphasic shock (two 5-ms pulses separated by 0.2 ms), delivered to coupled pairs of electrodes, to those using a sequential-biphasic shock (four 5-ms pulses separated by 0.2 ms) delivered to separate opposing electrodes, delivered from the same electrodes for both waveforms. In eight open-chest anesthetized pigs, four mesh electrodes (Medtronic TX-7, 6.5 cm2), were sutured on the epicardium of the anterior and posterior surfaces of each ventricle. Shocks were delivered from a 200-microF capacitor bank. Triplicate DFTs were obtained using each waveform in a randomized crossover design. Initial leading edge voltage (mean +/- SEM: 420 +/ 33 V vs 497 +/- 34 V; P < 0.05), initial peak current (4.8 +/- 0.4 A vs 13 +/- 1.1 A; P < 0.001), and delivered energy (16.9 +/- 2.6 J vs 30.4 +/- 5.3 J; P < 0.05) at the DFT were all significantly lower using sequential-biphasic shocks than those using single-biphasic shocks, respectively. We conclude that for direct heart defibrillation, it is worthwhile to combine sequential capability to biphasic shocks and deliver the remaining charge on the capacitor to the two different pathways. PMID- 9227757 TI - The effect of biphasic waveform tilt in transvenous atrial defibrillation. AB - Atrial defibrillation can be accomplished using low energy shocks and transvenous catheters. The biphasic waveform tilt required to achieve optimal atrial defibrillation thresholds (ADFTs) is, however, not known. The effect of single capacitor biphasic waveform tilt modification on ADFT was assessed in 20 patients. Following AF induction the defibrillation pulses were delivered between the catheters positioned in the coronary sinus and the right atrium. The single capacitor biphasic waveform shocks, delivered over the same pathways, consisted of 65% tilt (65/65 biphasic waveform) to produce an overall tilt of 88%, or 50% tilt (50/50 biphasic waveform) to produce an overall tilt of 75%. Although 65/65 biphasic waveform delivers more energy, the shorter duration 50/50 biphasic waveform reduced stored energy ADFT 21%, from 1.34 +/- 0.82 J with 65/65 biphasic to 1.06 +/- 0.81 J. These differences were not statistically significant. Nine patients had lower ADFT with 50/50 biphasic waveform while five patients had lower ADFT with 65/65 biphasic waveform. Equivalent reduction in ADFT was seen in the remaining six patients. The ADFT was 0.83 +/- 0.65 J when both tilts were considered. In conclusion, biphasic waveform tilt modification may affect the ADFT in an individual patient. The optimal biphasic waveform for ADFT is not known. PMID- 9227758 TI - The use of left ventricular epicardial surface velocity patterns as a marker of pacing site. AB - Ectopic ventricular foci were simulated at selected endocardial sites in 15 closed-chest canines using ventricular pacing. During this pacing, a noninvasive x-ray backscatter imaging technique was used to measure epicardial LV displacements at 5-ms intervals during the cardiac cycle. These displacement measurements were used to calculate epicardial surface velocities in each study and were presented as a time sequence of color coded velocity maps. Characteristic patterns in the timing and spatial propagation of LV surface velocities were noted for each pacing site, particularly during the expansion of the LV during isovolumic contraction and the inward motion of the LV during ejection. Average surface velocity maps for the 15 canines were computed for each pacing site. These average maps were used as standards for comparison with individual pacing studies to determine the probable site of pacing. Comparisons were made using a computer algorithm, based upon auto- and cross-correlation techniques in the time domain. This algorithm correctly identified pacing sites with sensitivities of RA 74%, LV 76%, RV 79%, and RVOT 77% and specificities of RA 98%, LV 96%, RV 90%, and RVOT 93%. The results show that this noninvasive mapping procedure has potential for identifying the location of an ectopic ventricular focus. PMID- 9227759 TI - Psychiatric morbidity and depressive symptomatology in patients with permanent pacemakers. AB - Implantation of a permanent pacemaker requires a psychological effort on the patient's part for adaptation in the acute term, and chronically, it restricts activities of the patient and may cause some psychiatric disturbances. To investigate psychiatric morbidity and depressive symptomatology of the patients with permanent pacemakers, 84 pacemaker patients were diagnosed using the DSM-III R criteria and depressive symptoms were determined by modified Hamilton Depression Rating Scale (mHDRS). Sixteen (19.1%) patients had been given a psychiatric diagnosis. The most frequent diagnoses were adjustment disorder (5.9%) and major depressive episode (4.7%). Nine patients (10.7%) were diagnosed as having clinical depression (mHDRS > or = 17). The mean score of mHDRS was 7.57 +/- 7.46, and the severity of depression was significantly higher in females. The most frequent symptoms are difficulties in work and activities (53.6%), psychic anxiety (48.8%), loss of energy (42.9%), and hypochondriasis and insomnia (39.3%). Depressed mood, psychic anxiety, loss of energy, loss of interest, insomnia, and hypochondriasis were significantly more frequent in females. Uneducated patients had a more significant loss of energy than educated patients. Depressed mood, psychic anxiety, and somatic concerns and symptoms were more frequent in patients with permanent pacemakers than in the general population. These symptoms, resembling mixed anxiety-depression disorder, were related to fears of having a permanent pacemaker, since our series were composed of uneducated patients who did not have enough knowledge about the device. PMID- 9227760 TI - Use of serotonin re-uptake inhibitors as primary therapy for carotid sinus hypersensitivity. AB - Carotid sinus syndrome (CSS) is a well-recognized cause of unexplained syncope in older patients, and may lead to significant morbidity related to trauma suffered during falls. Dual chamber pacing has been demonstrated to be efficacious in relieving symptoms due to bradycardia, but not the accompanying vasodepressor response. We report three patients with recurrent syncope due to a mixed type of CSS, who were treated with serotonin re-uptake inhibitors (SSRIs) alone, and were symptom-free after 4-5 weeks of therapy. The patients have remained symptom-free after more than 13 months of treatment. We conclude that SSRIs may be potentially useful in the treatment of CSS, and that the central mechanisms involved in CSS may be similar to those that result in neurocardiogenic syncope. PMID- 9227761 TI - Immediate and short-term reproducibility of the P wave signal-averaged electrocardiogram. AB - While abnormalities in the P wave SAECG have been associated with the occurrence of AF, its reproducibility has never been documented. The purpose of this study was to evaluate the immediate and short-term reproducibility of measurements from the P wave SAECG. P wave SAECGs were obtained using well-described techniques that utilize the QRS complex as the trigger and the P wave as template for averaging. In 28 subjects (8 controls, 11 with cardiac disease, 9 with prior AF), 3 P wave SAECGs were obtained: an initial study; on immediate reacquisition; and reacquisition after 4-5 days. Vector duration and RMS voltage of the terminal 20 ms of the P wave SAECG were measured and compared. The mean P wave duration was 152 +/- 14 ms on initial SAECG, 152 +/- 14 ms and 152 +/- 15 ms at immediate and short-term reacquisitions, respectively (both P = NS vs initial). The mean terminal RMS voltage was 6.4 +/- 6.0 mcV on initial SAECG, 6.4 +/- 5.9 mcV and 6.5 +/- 5.8 mcV at immediate and short-term reacquisitions, respectively (both P = NS vs initial). Linear regression analysis showed high reproducibility for both P wave duration (r = 0.94 for immediate and r = 0.96 for short-term reacquisition vs initial) but slightly less for terminal RMS voltage (r = 0.92 for immediate and r = 0.84 for short-term reacquisition vs initial). In subgroup analysis, P wave duration measurements were highly reproducible in controls, in subjects with cardiac disease, and in those with a history of AF. P wave duration was also reproducible for both males and females, as well as for subjects age > 65 years (r = 0.96 and 0.89 for immediate and short-term reacquisition, respectively). Terminal RMS voltage measurements were reproducible for controls, but less reproducible in other subgroups. In conclusion, P wave duration measurements on SAECG are reproducible when evaluated at immediate and short-term reacquisition regardless of age, sex, cardiac disease, or prior AF. Terminal RMS voltages were less reproducible, especially in patients with cardiac disease and/or prior AF. These findings may explain conflicting observations regarding the clinical utility of terminal P wave measurements. PMID- 9227762 TI - Influence of early coronary reperfusion on QT interval dispersion after acute myocardial infarction. AB - We studied the influence of early coronary reperfusion on QT interval dispersion in patients with acute myocardial infarction (MI). There were 54 males and 18 females with a mean age of 60 +/- 10 years. Of the 51 patients with recanalization of the infarct related vessel in the recovery phase, 28 (group A) had early coronary reperfusion (5.5 +/- 2.7 hours), 23 other patients (group B) were not confirmed with early coronary reperfusion. Twenty-one patients (group C) did not undergo recanalization of the infarct related vessel in the recovery phase. Corrected QT (QTc) maximum, QTc minimum, and QTc dispersion calculated as the difference between the maximum and minimum QTc intervals, were compared among these three groups at both acute and recovery phase. At the acute phase after MI, there were no significant differences in the QTc maximum, QTc minimum, QT dispersion, and QTc dispersion among these three groups. At the recovery phase after MI, there were also no significant differences in the QTc maximum and QTc minimum. However, there were significant differences in the QT dispersion (0.035 +/- 0.010 in group A, 0.049 +/- 0.015 in group B, and 0.061 +/- 0.031 s in group C, respectively; P = 0.0001), and QTc dispersion (0.038 +/- 0.012 in group A, 0.050 +/- 0.015 in group B, and 0.063 +/- 0.032 s in group C, respectively; P = 0.0003) among the three groups. Comparison of QTc dispersion between acute and recovery phase revealed significant reduction from acute to recovery phase in group A. The number of premature ventricular contraction was lower in group A and B than group C. In summary, early coronary reperfusion may reduce electrophysiological instability by reducing QT dispersion in the recovery phase after acute MI. PMID- 9227763 TI - Spontaneous accelerated junctional rhythm: an unusual but useful observation prior to radiofrequency catheter ablation for atrioventricular node reentrant tachycardia in young patients. AB - Between May 1990 and March 1995, 5 of 29 young patients (ages 4.2-25 years; median 14.1 years) undergoing RF ablation for atrioventricular node reentrant tachycardia (AVNRT) presented with spontaneous accelerated junctional rhythm (AJR) (CL = 500-750 ms), compared to 0 of 58 age matched controls undergoing RF ablation for a concealed AV accessory pathway (P = 0.004). In 3 of the 5 patients with AVNRT and AJR, junctional beats served as a trigger for reentry. During attempted slow pathway modification in the five patients with AVNRT and AJR, AVNRT continued to be inducible until the AJR was entirely eliminated or dramatically slowed. These 5 patients are tachycardia-free in followup (median 15 months; range 6-31 months) with only 1 of the 5 patients continuing to experience episodic AJR at rates slower than observed preablation. Episodic spontaneous AJR is statistically associated with AVNRT in young patients and can serve as a trigger for reentry. Successful modification of slow pathway conduction may be predicted by the elimination of AJR or its modulation to slower rates, suggesting that the rhythm is secondary to enhanced automaticity arising near or within the slow pathway. PMID- 9227764 TI - A patch in the pectoral position lowers defibrillation threshold. AB - Implantable pacemaker cardioverter defibrillators are now available with biphasic waveforms, which have been shown to markedly improve defibrillation thresholds (DFTs). However, in a number of patients the DFT remains high. Also, DFT may increase after implantation, especially if antiarrhythmic drugs are added. We report on the use of a subcutaneous patch in the pectoral position in 15 patients receiving a transvenous defibrillator as a method of easily reducing the DFT. A 660-mm2 patch electrode was placed beneath the generator in a pocket created on the pectoral fascia. The energy required for defibrillation was lowered by 56% on average, and the system impedance was lowered by a mean of 25%. This maneuver allowed all patients to undergo a successful implant with adequate safety margin. PMID- 9227766 TI - Regional wall motion during pacing for hypertrophic obstructive cardiomyopathy. AB - In order to assess the influence of right ventricular stimulation on LV contraction sequence in hypertrophic obstructive cardiomyopathy (HOCM), we performed a regional wall-motion analysis of the left ventricle by comparing normal His-Purkinje activation to pacing from the right ventricular apex. In 9 patients (5 males and 4 females, mean age 61 +/- 9 years) assessed after a mean pacing period of 12 weeks (range 7-24 weeks), AV sequential pacing induced a 52% reduction in maximal pressure gradient from 76 +/- 36 to 40 +/- 28 mmHg (P < 0.01) as determined by Doppler examination. Regional wall-motion analysis of the left ventricle was computed from digitized two-dimensional echocardiographic images by means of the area shrinking method. Pacing induced a significant reduction of total septal area shrinking from 25% +/- 17% to 12% +/- 17% (P < 0.005). A tendency toward paradoxical septal motion was observed in one patient only. The apex showed no significant variation. A 6% increase in area shrinking was observed at the posterior wall and lateral free wall, from 38% +/- 13% to 43% +/- 10% (P < 0.05). Pacing did not significantly alter the global ejection fraction. A direct correlation between the magnitude of subaortic pressure gradient reduction and that of septal motion changes was found in a majority of patients. In conclusion, dual chamber pacing reduces septal wall motion in patients with HOCM obstructive cardiomyopathy. This might be one of the mechanisms involved in the reduction of LV outflow tract obstruction. PMID- 9227765 TI - Sudden cerebral vasoconstriction during induced polymorphic ventricular tachycardia and fibrillation: further observations of a paradoxic response. AB - To determine the effect of induced polymorphic VT/VF on the cerebral circulation, transcranial Doppler (TCD) ultrasonography was used to prospectively assess changes in cerebral blood flow velocity during ICD implantation. Fourteen patients (13 men, 1 woman, mean age 58 +/- 20 years, range 34-74 years) who were survivors of an out of hospital cardiac arrest, were evaluated during routine ICD implantation. TCD ultrasonography was used to assess middle cerebral artery systolic velocity (Vs), diastolic velocity (Vd), pulsatility index (PI = Vs Vd/Vmean) and resistance index (RI = Vs-Vd/Vs) before, during, and after DFT testing with alternating current induction of polymorphic VT/VF. In each of the 14 patients studied, concomitant with the abrupt onset of hypotension, TCD sonography demonstrated a 33% +/- 28% decrease in diastolic velocity, a 42% +/- 28% increase in systolic velocity, a 190% +/- 141% increase in PI, and a 44% +/- 19% increase in RI. These findings reflect an increase in cerebrovascular resistance secondary to arteriolar vasoconstriction distal to the point of insonation of the middle cerebral artery. This response is paradoxic, as the expected response of the cerebral circulation to hypotension is vasodilation, but it is consistent with observations made in other acute hypotensive settings, such as tilt induced neurocardiogenic syncope. PMID- 9227767 TI - Distortion of intracardiac electrograms following defibrillator shocks for atrial tachyarrhythmias. AB - In three patients with a defibrillator system consisting of a Ventak P2 pulse generator and an Endotak C transvenous lead, we observed distortion of intracardiac electrograms following defibrillator shocks for atrial arrhythmias. There was a transient marked widening of the intracardiac ventricular complexes resembling ventricular tachycardia. This phenomenon should be recognized when evaluating arrhythmic episodes. PMID- 9227768 TI - Maturation of the sensed electrogram amplitude over time in a new subcutaneous implantable loop recorder. AB - The cause of recurrent syncope may be difficult to determine if the diagnosis is not established from initial noninvasive and invasive testing. Eighteen patients with recurrent syncope and negative tilt table and electrophysiological testing underwent implantation of a left pectoral subcutaneous loop recorder. This device "freezes" the preceding 7.5 or 15 minute rhythm strip after magnet application after spontaneous syncope. Baseline and follow-up electrograms were routinely recorded, and patients were followed until syncope recurred. Three patients had syncope within 1 month of implantation and were excluded from this report. Implantation electrogram amplitude was 250 +/- 124 microV and increased to 291 +/ 114 microV at 2-3 months, and increased further to 353 +/- 167 microV at 4-6 months (P < 0.001, ANOVA). Syncope recurred in 14 of the 15 patients. An arrhythmic basis for syncope was established (n = 7) or excluded (n = 7) in every patient who had recurrent syncope. All syncopal episodes were associated with diagnostic sensed electrograms. The increase in sensed electrogram amplitude over time suggests a maturation of the device-tissue interface. These results support the long-term viability of this implantable monitoring technique. PMID- 9227769 TI - High thresholds may alter end-of-life behavior in a dual chamber pacemaker. AB - We noted a series of 12 consecutive patients with a DDD Genisis pacemaker that showed an unexpected and a relatively rapid fall in battery voltage and output as these devices approached end-of-life (EOL). Twenty-one of 24 leads were Vitatron Helifix leads and there was a relatively high mean threshold (atrial 2.5 +/- 0.94 V; ventricular 2.9 +/- 0.65 V). These devices were replaced after 65 +/- 12 months. During the 9.3 +/- 3.5 months before replacement, a striking fall in voltage from 2.7 +/- 0.04 V to 2.49 +/- 0.05 V was seen. Battery impedance rose from 3 +/- 1.2 K omega to 10.2 +/- 4.3 K omega during this same period. We unexpectedly observed a marked difference between programmed and telemetered output for both atrial (50%) and ventricular leads (30%). A discrepancy between measured and telemetered magnet rate was also seen. Despite this relatively rapid fall in battery voltage, several of these devices did not meet the manufacturer's recommended replacement time (RRT) criteria by magnet rate or according to the projected RRT determined by the relationship of battery impedance to current drain. These data have implications for the selection of RRT and EOL criteria for this device. Magnet rate measured by surface ECG was the safest indicator for RRT. Follow-up for this pulse generator should be increased to every 2 months when battery impedance is > 2 KOhms or if there is a difference between programmed and measured output amplitude of more than 15%. The data also highlight the effect of combining high threshold leads with modern pacemakers with relatively "small" batteries as well as certain problems with telemetered data. PMID- 9227770 TI - Safety of slow pathway ablation in patients with long PR interval: further evidence of fast and slow pathway interaction. AB - Whether the presence of abnormal PR before selective slow pathway ablation for AV node reentrant tachycardia increased the risk of complete heart block remains controversial. We report our experience in seven patients with prolonged PR intervals undergoing catheter ablation for AV reentry tachycardia. Their mean age was 66 +/- 12 years; four patients were female and three were male. RF ablation was performed using an anatomically guided stepwise approach. In six patients, common type AV node reentry was induced and uncommon type was observed in the remaining patient. In all seven patients, successful selective slow pathway ablation was associated with no occurrence of complete heart block and was followed by shortening of the AH interval in five patients. In all seven patients, successful ablation was achieved at anterior sites (M1 in two patients and M2 in five patients). Despite AH shortening after ablation, the 1:1 AV conduction was prolonged after elimination of the slow pathway, excluding either sympathetic tone activation or parasympathetic denervation. In conclusion, selective slow pathway ablation can be performed safely in the majority of patients with prolonged PR interval before the procedure. Because successful ablation is achieved at anterior sites in most patients, careful selection and monitoring of catheter position is required. PMID- 9227771 TI - Dual device therapy in the setting of changing ICD technology: device-device interaction revisited. AB - This case report concerns an adverse device-device interaction between a replacement ICD and a dual chamber rate responsive pacemaker. It was observed that subtle changes in the design of sensing circuits between an older first generation ICD and the newer third-generation ICD device led to unexpected and dramatic changes in the interactive behavior of a dual device system. The new ICD was connected to chronically implanted hardware. The sensing behavior of the newer ICD included a shorter time constant in the decay of the automatic gain control function, resulting in triple sensing of both the atrial and ventricular paced stimuli and the evoked QRS complex. Physicians should be aware of new design changes in the future so as to anticipate such interactions. In the setting of rapidly changing technology, extra caution must be exercised when choosing to implant two devices in the same patient. PMID- 9227772 TI - Computation modes of multivariate positive predictive characteristics. AB - Multivariate receiver operator characteristics (ROCs) and positive predictive characteristics (PPCs), based on a combination of two or more clinical variables, are usually computed by varying dichotomy limits for each variable independently. This approach has a similar number of degrees of freedom (i.e., uses the same number of programmable parameters) as the approach which defines test positive cases based on a linear combination of all the clinical variables involved. Either approach can be implemented without any assumption about the underlying probability distributions by using an exhaustive computer search. Both approaches were compared in a demonstration study of predicting 2-year all cause mortality after acute myocardial infarction, based on applying various time- and spectral domain indices of signal-averaged ECGs from a research survey. The results showed that the optimum mode for the computation of ROCs and PPCs depends on the character of the particular data used. Therefore, in order to increase the precision of the retrospective multifactoral studies, both approaches to ROC and PPC computation should be used and compared in each individual investigation. PMID- 9227773 TI - Old issues, new challenges arise with the coming era of the electronic record and data exchange. PMID- 9227774 TI - Wide QRS complex tachycardia in the presence of preexcitation: a diagnostic challenge. PMID- 9227775 TI - Inappropriate discharges of an implantable cardioverter defibrillator secondary to automatic adjustable gain of atrial tachycardia. AB - Third generation implantable cardioverter defibrillators are capable of complex arrhythmia detection using sensing algorithms with automatic adjustable gain settings. We report a unique case where inappropriate sensing of atrial tachycardia in a patient with a His bundle ablation lead to satisfaction of ventricular fibrillation detection criteria. PMID- 9227776 TI - Inadequacy of qualitative implantable cardioverter defibrillator electrogram analysis to distinguish supraventricular from ventricular tachycardia due to electrogram changes during normally conducted complexes. AB - Stored electrograms (EGMs) recorded from ICD leads are used to evaluate the appropriateness of ICD therapies. Stored EGMs different from sinus have been interpreted as ventricular in origin. We present a patient with an ICD for VT who received multiple shocks for a tachycardia with a stored EGM different than sinus, suggesting VT. An electrophysiological study demonstrated EGMs different than sinus during atrial pacing and induced supraventricular arrhythmias. This case points out the limitations of stored EGMs and suggests complete electrophysiological study with analysis of EGMs during induced arrhythmias should be performed prior to discharge. PMID- 9227777 TI - Right ventricular radiofrequency ablation of ventricular tachycardia after myocardial infarction. AB - Radiofrequency transcatheter ablation of ventricular tachycardia in the setting of a prior myocardial infarction is typically performed with application of energy to the left ventricular endocardium. In this article, two cases are described in which successful radiofrequency transcatheter ablation of ventricular tachycardia occurred with energy delivery to the right ventricular septum after failed ablation attempts from the left ventricle. Both patients had tachycardias with a left bundle branch block morphology and markedly presystolic activity recorded from the right ventricular septum. Right ventricular septal activation mapping during ventricular tachycardia should be performed in patients with left bundle branch block tachycardia morphology and coronary artery disease to maximize efficacy of the catheter ablation procedure. PMID- 9227778 TI - Right hemidiaphragmatic twitching: a complication of bipolar atrial pacing. AB - A patient with an atrioventricular sequential pacemaker developed rhythmic contractions of the right hemidiaphragm. This was found to be the result of right phrenic nerve stimulation, which was directly related to the wide distance between the poles of an ELA atrial lead when pacing in bipolar mode. Pole separation in current atrial leads is discussed. PMID- 9227779 TI - Ten-year follow-up of a patient with pacemaker induced superior vena cava syndrome. AB - A woman with persistent pacemaker induced superior vena cava syndrome was stable for 10 years. Serial follow-up venography, however, demonstrated a continuous process of major vein occlusion and the development of collateral circulation, the effectiveness of which warrants a favorable prognosis in this pacemaker related syndrome. PMID- 9227780 TI - Implantation of dual chamber pacemaker in a patient with persistent left superior vena cava. AB - A 35-year-old patient underwent permanent pacemaker implantation because of symptomatic sinus bradycardia. During the procedure, persistent left superior vena cava was found. The ventricular lead crossed the tricuspid valve only after curving the stylet to form a loop in the right atrium (RA); subsequently, the curved stylet was changed to a straight one and the lead was positioned and screwed into the right ventricular apex. The atrial lead positioning was possible when the stylet was slightly curved and the lead could reach the anterior wall of the RA. At 18 months, a follow-up revealed normal pacemaker function and stable lead position. PMID- 9227781 TI - Unusual placement of a pacemaker pocket. AB - Because of infection and extensive radiation injury to the skin of the chest wall, a dual chamber pacemaker was implanted deep to the clavicle. The wound has been stable and comfortable for 1 year and pacemaker programming has been unimpaired. PMID- 9227782 TI - STIMAREC report. PMID- 9227783 TI - Should unipolar pacemaker leads be banned? PMID- 9227784 TI - Should unipolar pacemaker leads be banned? PMID- 9227785 TI - Should unipolar pacemaker leads be banned? PMID- 9227786 TI - Scintimammography with Tc-99m sestamibi. AB - Mammography and physical breast examination are currently the most frequently and recognized screening tools for detection of breast carcinoma. These methods have been proved successful for early detection of breast cancer. Considering the 85% sensitivity associated with combined mammography and physical examination and a low positive predictive value of 20%-30% for diagnosis of breast carcinoma, there is a critical need for a more accurate, noninvasive imaging test to improve the sensitivity and specificity of mammography (7, 19, 20). Since early 1992, we have studied over 1200 women with clinically and/or mammographic abnormalities prior to breast biopsy and/or fine needle aspiration cytology of the breast. We have evaluated the role of Tc 99m Sestamibi as a complimentary procedure to conventional mammography in detection of breast carcinoma. The preliminary results of our studies have been published elsewhere (14, 17, 18). DuPont Merck Pharmaceutical Company in the USA on that basis, determined to conduct a multicenter clinical trial for the role of this radiopharmaceutical for the diagnosis of breast carcinoma in women with mammographically and/or clinically palpable abnormalities. This study was conducted at 42 institutions throughout the United States and Canada enrolling 673 women who where otherwise scheduled for breast biopsy and/or mastectomy. The preliminary results of this trial in both palpable and nonpalpable breast abnormalities are encouraging (24). Our most recent study on 157 women (mean age 47.9 years +/- 10.2) with 164 lesions with indications for histologic and cytologic analysis who underwent scintimammography with Tc 99m Sestamibi demonstrated the sensitivity of 92.3% and the specificity 87.5% (15). We have concluded that Scintimammography with Tc 99m Sestamibi can be used in conjunction with mammography to improve its specificity. PMID- 9227787 TI - Scintimammography using Tc-99m tetrofosmin. AB - Breast cancer, the most common malignancy in women, still poses a challenge to diagnostic procedures and therapy. Despite low specificity routine mammography is the method of choice to screen women for breast cancer. In the last years other additional diagnostic procedures such as high frequency ultrasonography (US) and especially magnetic resonance imaging (MRI) have improved breast cancer diagnosis. However, all these imaging methods are lacking in specificity which makes biopsy or surgery necessary. The purpose of our study was to evaluate prospectively the sensitivity, specificity, PPV and NPV of scintimammography with a new cationic complex Tc-99m tetrofosmin in patients with suspicious mammographic lesions. One hundred and thirty seven patients in whom mammography and/or high resolution ultrasonography (10 MHz) revealed suspicious breast lesions were studied with Tc-99m tetrofosmin scintimammography. In 84 of them biopsy and/or surgery was performed for histological evaluation. After intravenous injection of 555 MBq Tc-99m tetrofosmin planar images in anterior and lateral projections (5 min.p.i.) and SPECT imaging including 3-D-reconstruction (20 min.p.i.) were performed. Scintimammography was evaluated as negative, equivocal (+), probably (+2) or definitely (+3) positive. Planar scintimammography with Tc-99m tetrofosmin was negative in 46 patients (43 true negative-f.n; 3 false negative-f.n.) and positive in 38 patients (27 true positive-t.p.; 11 false positive-t.p.). Using SPECT imaging Tc-99m scintimammography was negative in 43 cases (41 t.n.; 2f.n.) and positive in 41 cases (28 t.p.; 13 t.p). Sensitivity of Tc-99m tetrofosmin scintimammography in this prospective study was 90%, specificity 80%, PPV 71% and NPV 93% for planar imaging and 93%, 76%, 68% and 95% for SPECT, respectively. Scintimammographic results in patients with suspicious breast lesion show, that Tc-99m tetrofosmin accumulates in breast cancer as well as in some fibroadenoma with high cellularity. However, the high NPV of 93 and 95% respectively excludes breast cancer in suspicious mammographic lesions to a very high degree and therefore reduces the need of biopsy and/or surgery in most of these patients. PMID- 9227788 TI - Studies of breast cancer imaging with radiolabeled antibodies to carcinoembryonic antigen. Immunomedics Breast Cancer Study Group. AB - Multiple imaging methods are currently used to detect the presence, location, and extent of breast cancer, either at initial presentation or during follow-up. Even at initial diagnosis, more specific methods are needed to complement the high sensitivity of mammography in the detection and diagnosis of mammary carcinoma. Therefore, the current study evaluates the potential use of a new monoclonal antibody-based imaging agent, arcitumomab (CEA-Scan), which comprises an anti carcinoembryonic antigen monoclonal antibody Fab' fragment labeled directly with technetium-99m, in the detection of primary, recurrent, or metastatic mammary cancer. A prospective, non-randomized, open-label, multicenter trial was conducted in 123 women 100 with primary breast cancer confirmed by histology or cytology, and 23 with a prior history and who were under evaluation for possible recurrence or metastasis. A 1-mg dose of arcitumomab labeled with 20-25 mCi Tc 99m was injected i.v., and planar images were acquired at 5-8 hr, as well as single-photon emission computed tomography. Additional planar scanning was performed optionally at 18-24 hr. The imaging studies were interpreted as positive if there was abnormal focal activity, and were correlated, wherever possible, with histopathology or conventional imaging tests. In 78 evaluable patients with primary breast carcinoma arcitumomab showed an imaging sensitivity and specificity of 82% and 88%, respectively, even detecting lesions < 1 cm in 61% of the cases. Among 58 patients evaluable for axillary node involvement, 30 were positive by histopathology, of which arcitumomab was correct in 19 with cancer (63% sensitivity) and in 25 of 28 without cancer (89% specificity). Among 88 primary operable patients, arcitumomab showed additional sites of focal uptake outside of the breasts and axillae in 10, or 11%. In a total of 19 breast cancer patients with known or suspected recurrence or metastatic disease, 13, or 68%, showed one or more focal sites of uptake, involving the typical sites of spread of mammary cancer. No significant adverse effects or induction of human antimouse antibodies were observed in these patients. Arcitumomab immunoscintigraphy is a simple, same-day, and safe new imaging modality for the detection of sites of breast cancer, including lesions < 1 cm, with a reasonably high rate of sensitivity and specificity. It may be used to complement mammography in the differentiation of malignant from benign disease. PMID- 9227789 TI - The role of Na+ channels in twitch generation during exposure of the frog rectus abdominis to Ca-free Ringer solution with Na2EDTA. AB - The aim of the study was to investigate whether Na+ channels play a role in the twitch component of the response of the isolated frog rectus abdominis to Ca(2+) free Ringer solution with 0.2 mM Na2EDTA by using tetrodotoxin and some other well known drugs that exhibit a blocking action on Na+ channels. In the presence of 5 x 10(-7) M tetrodotoxin, the twitch component, measured isotonically, disappeared. Although 10(-7) M d-tubocurarine was found to be ineffective, a complete blockage of twitch amplitude was observed at 5 x 10(-6) M concentration of the drug. The inhibitory action of d-tubocurarine on twitch response was not antagonized by 10(-6) and 10(-5) M carbachol. Propranolol (10(-6) - 10(-5) M), lidocaine (2 x 10(-6) - 10(-5) M), quinine (10(-6) - 2 x 10(-5) M) and quinidine (10(-6) - 2 x 10(-5) M) inhibited maximal twitch amplitude in a concentration dependent manner. These findings strongly suggest that activation of tetrodotoxin sensitive Na+ channel may play a primary role at twitch generation during exposure of the frog rectus abdominis to Ca(2+)-free Ringer solution with Na2 EDTA. PMID- 9227790 TI - Growth inhibitory effects of antifolates against an adriamycin-resistant human small cell lung cancer cell line. AB - We have established an Adriamycin (ADM)-resistant small cell lung cancer (SCLC) cell line, SBC-3/ADM 100, which shows multifactorial mechanisms of resistance to ADM, such as over-expression of P-glycoprotein, an enhanced detoxifying system and a decrease in topoisomerase II activity. In the present study, we confirmed that SBC-3/ADM 100 showed collateral sensitivity to methotrexate and TNP-351, a new antifolate, though this cell line showed a typical multidrug resistance (MDR) pattern. We also demonstrated a faster uptake and higher accumulation (1.3-fold) of TNP-351 in the SBC-3/ADM 100 cells than those in the parent SBC-3 cells. These results explain one of the mechanisms for collateral sensitivity in the resistant cells. Furthermore, this cell line was found to have no cross-resistance to edatrexate and minimal cross-resistance to trimetrexate, 254-S (cisplatin analog), 5-fluorouacil and 4-hydroperoxyifosfamide. These drugs will have clinical importance in patients with SCLC who were previously treated with an ADM containing regimen. Thus, antifolates, especially TNP-351 and edatrexate, can be expected to eradicate residual multidrug resistant SCLC cells selected by ADM. PMID- 9227791 TI - The effect of iron supplementation on GSH levels, GSH-Px, and SOD activities of erythrocytes in L-thyroxine administration. AB - Our aim was to study the effect of iron supplementation on the following aspects of erythrocyte metabolism in experimental hyperthyroidism: glutathione (GSH) levels, glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) activities. Hyperthyroidism induced by L-thyroxine administrations significantly raised erythrocyte GSH, GSH-Px and SOD levels of the rats (P < 0.001). Likewise, we observed that iron supplementation induced significant rises in erythrocyte GSH, GSH-Px and SOD levels (P < 0.001) as compared with the control group. The erythrocyte GSH, GSH-Px and SOD levels of hyperthyroidism-induced iron supplemented animals were significantly higher when compared with either the iron supplemented group (P < 0.001) or the only L-thyroxine-administered hyperthyroid group (P < 0.001, P < 0.05, P < 0.01, respectively). The results of this study show that L-thyroxine administration and/or iron supplementation increases GSH, GSH-Px and SOD levels of erythrocytes. PMID- 9227793 TI - Malignant lymphoma induction in rabbits by oral inoculation of crude virus fraction prepared from Ts-B6 cells (cynomolgus B-lymphoblastoid cells harboring Epstein-Barr virus-related simian herpesvirus). AB - Malignant lymphoma was induced in Japanese (JWY), New Zealand (NZY) and Dutch (DUY) white rabbits by oral spray of cell-free pellets of culture fluid (crude virus fraction) of Ts-B6 cells (cynomolgus monkey B-lymphoblastoid cells harboring Epstein Barr virus-related simian herpesvirus or Cyno-EBV). Nine of 11 inoculated rabbits developed malignant lymphomas within 42-160 days after oral inoculation (JWY, 2/3; NZY, 5/6; DUY, 2/2). In contrast, none of the control rabbits inoculated in the same fashion with B95-8 (EBV-producing marmoset cell line) cell-free pellets developed malignant lymphoma. Most rabbits showed increased anti-VCA IgG and anti-EA-DR IgG antibody titers after inoculation by oral spray of Ts-B6 cell-free pellets. EBV-encoded RNA-1 was revealed in the tumor cells by in situ hybridization. EBV DNA was detected in the rabbit peripheral blood leukocytes (PBL) by polymerase chain reaction; the earliest positive result was obtained only two days after oral inoculation. These data suggest that orally administered Cyno-EBV in Ts-B6 cells infects PBL and then induces malignant lymphoma in rabbits. The availability of this animal model promises to clarify the role of EBV in human lymphoma and provides a means for studying prophylactic and therapeutic regimens. PMID- 9227792 TI - Effects of simulated microgravity on human osteoblast-like cells in culture. AB - Physiological strain plays an important role in maintaining the normal function and metabolism of bone cells. It is well know that the mineral content of astronauts' bones decreases during spaceflight. Thus, gravity is one of the important factors in the musculoskeletal system. The vector-free horizontal clinostat has been used to simulate conditions of microgravity for examining such effects on cells in culture. We analyzed the effects of simulated microgravity using a horizontal clinostat on cultured osteoblast-like cells (HuO9 cell line). Total cellular protein, which was measured as an indication of cell proliferation, was not significantly inhibited under simulated microgravity conditions. No morphological changes were detected under microgravity conditions by phase-contrast microscopy. However, the alkaline phosphatase (ALP) activity and osteocalcin production of the HuO9 cells decreased significantly under microgravity conditions. Our data indicate that simulated microgravity directly inhibits some differentiation phenotypes and some functions of osteoblasts. On the other hand, the addition of 1,25-dihydroxyvitamin D3 (1,25-(OH)2-D3) increased ALP activity under simulated microgravity conditions, although the total activity of ALP in the cells treated with 1,25-(OH)2-D3 was still lower under simulated microgravity conditions than that in the control cells. However, the cells under simulated microgravity conditions showed a greater enhancement of ALP activity by treatment with 1,25-(OH)2-D3. PMID- 9227794 TI - Gene transfection by cationic liposomes: comparison of the transfection efficiency of liposomes prepared from various positively charged lipids. AB - We compared the transfection efficiency of four types of positively charged liposomes composed of (i) N-(alpha-trimethylammonioacetyl)-didodecyl-D-glutamate chloride (TMAG), dilauroylphosphatidylcholine (DLPC), and dioleoylphosphatidylethanolamine (DOPE) (1:2:2 molar ratio); (ii) 3 beta [N-(N', N'-dimethylaminoethane)-carbamoyl] cholesterol (DC-Chol) and DOPE (3:2 molar ratio); (iii) dimethyldioctadecylammonium bromide (DDAB) and DOPE (1:2.2 molar ratio); (iv) N-[1-(2,3-dioleyloxy) propyl]-N,N,N-trimethylammonium chloride (DOTMA) and DOPE (1:1, w/w; lipofectin). Luciferase gene was used as a reporter gene. Among the cationic lipsomes used, the liposomes composed of TMAG, DOPE and DLPC showed a much higher efficiency of plasmid DNA entrapment than the other cationic liposomes tested. In the absence of serum, the cationic multilamellar vesicles (MLV) and small unilamellar vesicles (SUV) composed of TMAG, DOPE and DLPC gave highly efficient transfection. On the other hand, MLV, dehydration rehydration vesicles (DRV), and SUV liposomes prepared with the mixtures of DC Chol and DOPE showed similar levels of transfection efficiency. However, the cationic liposomes composed of DDAB and DOPE showed inferior efficiency, whether in the form of DRV, SUV or MLV. The transfection efficiency of lipofectin was also low. In the presence of serum, on the other hand, a considerable (about 30 50%) amount of transfection activity was still observed at 10% fetal calf serum in the cationic MLV and SUV composed of TMAG, DOPE and DLPC. Cationic MLV, composed of TMAG, DOPE and DLPC, Cationic MLV, composed of TMAG, DOPE and DLPC, can transfect plasmid DNA, not only in the adherent cell lines but also, in the suspension cell lines. These findings indicate that the transfection efficiency of cationic liposomes is affected by the lipid composition, the type of liposome, or the presence or absence of serum. They also indicate that the cationic liposomes containing TMAG, DOPE and DLPC are efficient vectors for gene transfer into cells. PMID- 9227795 TI - Percutaneous release for trigger finger in idiopathic and hemodialysis patients. AB - Sixty-seven trigger fingers of 58 idiopathic and hemodialysis patients were treated by percutaneous A1-pulley release technique. Severity of triggering was classified into five grades for treatment selection and prediction of possible results. Results were excellent in 41 fingers, good in 9, fair in 7, and poor in 10, requiring additional treatment. The results of the lower grades were better, and those of the higher grades were poor. Excellent or good results appeared to depend on the proper selection of the patients according to the grading system and confirmation of triggering disappearance just after the release. There were neither infections nor neuro-vascular deficits after treatment. Compared to conventional open release, this treatment was found to be more useful from the standpoints of ease and safety of the technique, and the patients' quick return to normal life. PMID- 9227796 TI - Surgery of gastric cancer in patients over 80 years old. AB - A retrospective study on postoperative complications and factors affecting prognosis was performed on elderly patients with gastric cancer. We studied the correlation of age, pathological depth, preoperative laboratory data, physical status, duration of surgery, volume of blood loss, blood transfusion, curability, and extent of lymph node dissection to postoperative complications and prognosis in 47 patients with gastric cancer over 80 years old. Preoperative function of lung and liver frequently showed abnormal data. Postoperative complications were noted in 47% of patients, especially in the pulmonary system, liver and heart. Curability and extent of lymph node dissection were the significant factor affecting survival. Some mortalities caused by initial malignancy were recognized in the conservative lymph node dissection in the stage I. The incidence of postoperative complications was not significantly different according to extent of lymph node dissection. Blood transfusion was the only significant factor for the incidence of postoperative complication. The most frequent cause of death was the initial malignancy. We recommend that a low grade lymph node dissection should not be readily chosen for elderly patients in early cases. PMID- 9227797 TI - Cross-reactivity to olive tree pollen and orchard grass pollen in patients with pollinosis. AB - We studied 92 patients with allergic rhinitis in Syodoshima, Japan, during the pollen season between April and June to evaluate the cross-reactivity to different antigens, including pollen from the olive tree (Olea europaea) and orchard grass (Dactylis glomerata). Olive tree pollen was present in the atmosphere for 23 days, from May 19 to June 12, 1994. Specific IgE antibodies for olive tree pollen antigen were present in 21 (26.9%) of the 78 patients with allergic rhinitis. Nine (24.3%) of the 37 patients with allergic rhinitis exhibited positive skin reactivity to an extract of olive tree pollen. Fifteen (88.2%) of the 17 patients who had IgE reactivity in their sera to olive tree pollen antigen demonstrated allergic reactions to an extract of olive tree pollen. Specific IgE antibodies for orchard grass pollen antigen were present in 43 (48.3%) of the 89 patients with allergic rhinitis and 20 (95.2%) of the 21 patients who had IgE reactivity in their sera to olive tree pollen antigen. The inhibition test using the CAP System revealed that the reactivity of the IgE antibody specific for olive tree pollen antigen was inhibited dose-dependently by an extract of orchard grass pollen. These findings show that there is a reaction in some patients with grass (Gramineae) pollinosis that might be induced by olive tree pollen. PMID- 9227798 TI - Post treatment sensitivity studies with the ParaSight-F test for malaria diagnosis in Zimbabwe. AB - The post-treatment diagnostic performance of the Plasmodium falciparum histidine rich protein (HRP-II) antigen detection test, ParaSight-F test, was assessed on 55 falciparum malaria cases treated with chloroquine during in vivo drug sensitivity studies. The post-treatment sensitivity of the test remained high, except for an insignificant decline on day 1. However, specificity dropped sharply by day 1, subsequently increasing linearly with time to satisfactory values by day 10. As expected, from its inverse relationship to specificity, the false positive rate was high on day 1 and decreased linearly to low level by day 10. The temporary increase in false positive rate-following treatment was due to persistent parasite antigen, rather than subpatent parasitaemia. Thus findings showed that positive readings by the test within 10 days post-treatment may occur in cured cases and will not necessarily imply treatment failure. Furthermore it will be important to take patient antimalarial history into consideration during routine usage of the test for malaria diagnosis. The trend of Youden's J-index for the ParaSight-F test showed that from 10 days post-treatment, the test was generally reliable, with positive readings indicating active infection. It was concluded that the ParaSight-F test was not only valuable at confirming malaria diagnosis on clinical cases in seasonal transmission areas, but had potential for application to detect recrudescent infections within 2 weeks of chloroquine treatment. PMID- 9227800 TI - In vitro trypanocidal activity of some rare Tanzanian medicinal plants. AB - Extracts prepared from 15 rare Tanzanian medicinal plants were tested for their in vitro antitrypanosomal activity against human pathogenic trypanosomes (Trypanosoma brucei rhodesiense). Of 37 extracts investigated, two were found to have strong activity with IC50 values below 1 microgram/ml and ten extracts revealed promising activities with IC50 values between 1-5 micrograms/ml. PMID- 9227799 TI - Evaluation of alkaline phosphatase immunoassay and comparison with other diagnostic methods in areas of low transmission of schistosomiasis. AB - The alkaline phosphatase immunoassay (APIA) is an antibody detection technique which permits the diagnosis of schistosomiasis using a butanolic extract preparation from adult worms. APIA has demonstrated high sensitivity and specificity in previous reports with well characterized human sera. Its potential as a diagnostic tool for epidemiological surveillance was assessed in comparison with three other diagnostic tests: stool examination, ELISA with soluble egg antigen (SEA) and the circumoval precipitin test (COPT). APIA was 100% specific in an area without Schistosoma mansoni transmission and had 89% sensitivity in an endemic area where 69% of the infected subjects excreted less than 100 eggs g of faeces. It was found to be less sensitive in children under 5 years of age who were positive by the COPT test. APIA can be applied as an initial screening test, based on its high sensitivity, specificity, absence of cross-reactivity with intestinal parasites and the fact that it is a technique suitable for use in epidemiological surveillance. PMID- 9227801 TI - Enhanced detection of Schistosoma circulating antigens by testing 1 ml urine samples using immunomagnetic beads. AB - The potential advantage of increasing the sample volume for enhanced detection of two Schistosoma circulating antigens in urine has been evaluated. Two different, monoclonal antibody based, immunomagnetic beads assays detecting circulating anodic antigen (CAA) and circulating soluble egg antigen (CSEA), respectively, were investigated. By increasing the sample volume to 1.0 ml, the lower detection limit of antigen dilution series was improved six eight-fold in both immunomagnetic beads assays. Indeed, an improvement could be shown when testing the urine of Schistosoma-infected individuals with these magnetic beads assays. To compare the sensitivity of the immunomagnetic beads method with the standard ELISA, urine was experimentally adjusted to mirror low natural infection intensities. CSEA-positive urine samples were adjusted to levels below the detection limit of the corresponding ELISA and indeed were still found positive by the CSEA immunomagnetic beads assay. These results demonstrate that by using immunomagnetic beads, an assay method which can utilise larger sample volumes than an ELISA, enhanced detection can be achieved. This might be particularly valuable in areas with low prevalence of schistosomiasis or for the assessment of cure after chemotherapy. PMID- 9227802 TI - Schistosomiasis and the use of indigenous plant molluscicides: a rural South African perspective. AB - In the last decade plant molluscicides have received considerable attention in the search for cheaper alternatives to chemotherapy and synthetic molluscicides in schistosomiasis control. The attraction of a locally grown molluscicidal plant is based on the development of a philosophy of self-reliance and community involvement. This approach is dependent on community recognition of the infection as a public health problem and their acceptance of proposed control measures. The objectives of this study were: (i) firstly, to assess the knowledge of schistosomiasis in a rural community and their attitude to the use of indigenous plant molluscicides; (ii) secondly, to assess the prevalence and intensity of infection in relation to its severity as perceived within the community. Study sites were located at Mtwalume (KwaZulu-Natal, South Africa). Sixty-nine community members were interviewed during six focus-group interviews and two depth interviews. Urine and stool samples (354 and 306, respectively) from children and young adults (2-25 years old) were analysed for Helminth and Protozoal infections. Results indicate that despite a poor understanding of schistosomiasis, it is a primary health concern for those dependent on river water for their water requirements. Concern for schistosomiasis is indeed matched by a prevalence of 75.14% for Schistosoma haematobium. Oral antischistosomal drugs are inaccessible primarily due to the cost of transport and secondarily, due to the cost of treatment. The concept of molluscicidal control, as an alternative, was enthusiastically received by all respondents. PMID- 9227803 TI - Detrimental effect of nitric oxide on Trypanosoma cruzi and Leishmania major like cells. AB - The production of nitric oxide (NO) by activated macrophages has been reported to be a non-specific immune-effect mechanism against several parasites. In this work we investigate whether the NO has a detrimental effect on the intracellular parasites of the genus Leishmania and as well as Trypanosoma cruzi. This was assessed by co-cultivating infective Leishmania promastigotes and T. cruzi trypomastigotes and non-infective T. cruzi epimastigotes forms of the parasites in the presence of the NO releasing molecule, S-nitroso-N-acetyl-DL-penicillamine (SNAP). We demonstrate that the NO has the ability to inhibits the growth of all parasites in a concentration dependent manner. In addition, by analysing purified protein and cell homogenates of L. major (promastigotes) and T. cruzi (epimastigotes and trypomastigotes) we demonstrated that the NO may regulate the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) activity of both parasites. PMID- 9227804 TI - Effect of a low protein diet on the energy metabolism of growing chickens. AB - Two slaughter experiments were carried out to determine whether the protein content of the diet has an influence upon the efficiency of utilization of ME in fast growing chickens. A normal-protein diet (NPD, 204 g CP/kg DM: 14.7 MJ ME/kg DM) based on soybean meal as the sole source of protein was given at four different levels of intake (ad libitum or restricted at about 90, 65 and 40% ad lib) to 10-d-old animals for 2 weeks. In a parallel experiment the chickens were fed ad libitum a low protein diet (LPD, 66 g CP/kg DM: 15.0 MJ ME/kg DM) based on soybean meal. The intake of metabolizable energy ranged from 1675 to 777 and 1770 to 832 kJ/kgW0.75 per day for NPD and LPD treatments, respectively. Mean values of energy retention, gross efficiency of energy utilization and energy retained as protein were significantly (P < .05) lower and heat production (expressed as both kJ/kgW0.75 per day and kJ/kg body protein content0.75 per day) was significantly higher (P < .05) for the chickens fed on LPD. These findings support the concept of dietary-induced thermogenesis in response to reductions in dietary protein concentration. It is concluded that the increased heat production found in the birds fed on the low-protein diet can be explained by both an increase in energy requirements for maintenance (MEm) and a sharp decrease in the efficiency of utilization of ME of growth (k(g)). PMID- 9227805 TI - An investigation into the variability of extract viscosity of wheat-relationship with the content of non-starch-polysaccharide fractions and metabolisable energy for broiler chickens. AB - The in vitro extract-viscosity and the content of non-starch-polysaccharides were investigated in 34 defined wheat varieties grown at 5 locations each. Both, wheat genotype as well as growing location clearly influenced the viscosity of soluble extract from wheat. Furthermore, the content of non-starch-polysaccharides (soluble/total) and pentosans (soluble/total) were determined in 13 wheat varieties each grown at two locations. Soluble pentosan contents were highly positively correlated with extract viscosity of wheat at the locations Hayn (r = 0.86) and Biendorf (r = 0.90). The classical apparent metabolisable energy of 5 wheat samples having different extract viscosities was assessed. The AMEN values ranged from 14.0 to 14.6 MJ/kg DM and were significant negatively correlated to content of soluble arabinoxylans (r = 0.67) and to the extract viscosity (r = 0.83). Furthermore, the viscosity of jejunal (4.0 to 22.8 mPas) and ileal (13.1 to 78.0 mPas) digesta exhibited a clear relationship with soluble pentosan contents and extract viscosity. Under the conditions applied in this study the technique of extract viscosity measurement can predict the AME. PMID- 9227806 TI - Evaluation of the non invasive TOBEC (total body electrical conductivity) procedure for prediction of chemical components of male broilers with special consideration of dietary protein level. AB - TOBEC (total body electrical conductivity) measurement as a non invasive procedure for the estimation of body chemical composition was used to calculate calibration curves for the prediction of crude protein mass (CPM), crude water mass (CWM), crude ash mass (CAM) and fat-free mass (FFM) of male broiler chickens. A growth experiment with 3 protein levels (130, 230 and 330 g CP/kg diet, isoenergetic with 13.3 MJ AMEN/kg) was combined with TOBEC measurements and body chemical analysis in order to obtain the values necessary for calibration. A total of 196 TOBEC measurements and body chemical analysis were undertaken in time intervals of two days beginning with batch until day 17 of age. Different dietary protein levels resulted in marked differences in body weights and body chemical compositions but in similar TOBEC-responses for a given mass of FFM, CPM, CAM or CWM. Values for birds fed a diet with 130 g CP/kg diet tended to be more variable. Linear broken relationships were found between FFM, CPM, CAM and CWM, respectively and TOBEC values (E#). A set of different regression equations is given and yielded high proportions of variance accounted for the piece wise regression model (R2 ranged from 0.83 to 0.99). In spite of these high determinations the prediction of crude fat mass (CFM) by subtracting the FFM from the body weight resulted in most cases in weak determinations between observed and predicted CFM (R2 ranged from 0.38 to 0.86). The highest R2 was observed when the E# was expressed per unit metabolic body weight to the power of 0.67 and regressed on FFM expressed to the same power. In conclusion, FFM, CPM and CWM may be predicted reasonably well by TOBEC. However, these high determinations are not high enough to predict CFM accurately. In addition, the application of such regressions to an individual bird seems to be impossible. Assessment for groups of animals should be possible if errors of estimation, standard deviations and differences to be detected are taken into account in the calculation of the number of birds necessary for the TOBEC measurements. PMID- 9227807 TI - [Influence of the ration content on the fecal phosphorus excretion of growing bulls varying in body weight]. AB - Five smaller (body weight: 227.9 +/- 23.7 kg) and large growing bulls (435.2 +/- 14.3 kg per animal) each were fed with rations rich (chopped wheat straw:concentrate = 1:1) or poor in roughage (straw: concentrate = 1:4). Animals were kept in balance cages; 20 days adaptation period were followed by 10 days collection period. Body weight of bulls did not significantly influence apparent digestibility of rations. digestibility of organic matter of fibre rich ration was significantly lower (66.8%) than those of concentrate rich ration (74.4%). Feeding and body weight did not significantly influence metabolic parameters of mineral status. The fecal P-excretion amounted to 0.94 and 1.08 g per kg DMI in bulls fed with rations rich and poor in roughage. No influence of body weight was measured. Feeding and body weight did not significantly influence fecal P excretion per kg DOMI (between 1.44 and 1.58 g P/kg DOM). For calculation of P requirements for growing cattle fecal P-excretion amounted to 1.0 or 1.5 g P/kg DM and DOMI, respectively. PMID- 9227808 TI - Rumen dry matter and crude protein degradability of extracted or untreated oilseeds and Leucaena leucocephala leaves. AB - A study was undertaken to determine the rumen DM and CP degradability characteristics of soyabean, canola seed, peanut, palm kernel and Leucaena leucocephala leaves. The oilseeds were either treated with n-hexane to extract the fat or left untreated. Nylon bags were incubated in each of four rumen cannulated sheep for 0, 2, 4, 6, 12, 24 and 48 h. Animals were fed on a diet consisting of meadow hay (ad libitum) and 150 g of concentrate twice daily. Fat extraction caused a decrease (P < or = 0.05) in DM disappearance of soyabean at 0, 2, 4, 6 and 12 h and of peanuts at all incubation times. CP disappearance from peanuts was reduced (P < or = 0.05) as a result of fat extraction at 0, 2, 4, 6 and 12 h. Fat extraction of canola seed increased CP disappearance at 0, 2, 4, 6 and 24 h (P < or = 0.05). However, in the case of defatted canola seed, an increase in DM disappearance (P < or = 0.05) was observed in the first 4 incubation times and a decrease (P < or = 0.05) in the later times. Fat extraction increased (P < or = 0.05) DM disappearance of palm kernel at 0 and 48 h, but reduced it at 4, 6 and 24 h. CP disappearance of palm kernel was improved by treatment (P < or = 0.05) at 0, 4, 24 and 48 h and decreased at 12 h. In the case of palm kernel the largest differences in DM and CP disappearance occurred between the 24 and 48 h incubation times. Degradability characteristics for DM and CP of full-fat soyabean, canola seed and peanut were comparable to those of the full fat samples. Effective DM degradability of soyabean, canola seed and peanuts was 72.2 and 71.9; 74.1 and 66.8; and 85.9 and 70.8 for full fat and extracted feeds, respectively. Effective CP degradability was similar in all oilseeds with the exception of the extracted canola seed. Therefore, the incorporation of full-fat soyabean, canola seed and peanut into ruminant rations can be considered as a means of increasing the energy balance. Both palm kernel DM and CP degradabilities were characterized by slow rates of degradation by negative values "b". This suggests the predominance of microbial colonization over disappearance during incubation. DM and CP disappearance of Leucaena leucocephala leaves originating from Cuba were lower than those from Nigeria. Degradability characteristics for CP and DM of Cuban leucaena leaves showed that the linear model resulted in a better fit than the exponential one. PMID- 9227809 TI - Chronic renal disease and cardiovascular complications: inevitable or preventable? Now is the time for some intervention studies. PMID- 9227810 TI - Can we reduce acute asthma attendances to hospital emergency departments? PMID- 9227811 TI - Risk factors for repeat attendance at hospital emergency departments among adults and children with asthma. AB - BACKGROUND: Acute asthma attacks are frequent causes of attendance at hospital Emergency Departments (EDs) and a subgroup of these patients repeatedly present for such treatment. AIMS: This study sought to characterise patients who were repeat attenders at EDs, to assist the targetting of appropriate future interventions aimed at reducing avoidable presentation. METHODS: A cross sectional survey was undertaken of patients presenting with an asthma attack to the EDs of six teaching hospitals in Adelaide, South Australia between 14 May and 30 June 1994. Patients were interviewed within six weeks of their attendance about aspects of their asthma history, severity, medications, self-management, attitudes and environment. Repeat attenders, defined as two or more visits over the course of the preceding year, were compared with those who reportedly attended on one occasion only, using logistic regression analyses. RESULTS: Sixty two per cent of 272 patients aged under 15 years and 40% of 165 patients aged 15 years or more reported having attended two or more times over the course of the preceding year. Among adults, the variables independently associated with repeat attendance principally related to asthma severity. Among children, repeat attendance was associated with parental attitudinal variables relating to appraisal of their child's asthma severity, management of asthma attacks and parental worry. CONCLUSIONS: The factors underlying repeated presentations at EDs differ between adults and children and interventions to minimise avoidable presentation will require different emphasis for these patient subgroups. PMID- 9227812 TI - High density lipoprotein (HDL) particle composition in patients with end stage renal failure (ESRF) on chronic dialysis. AB - BACKGROUND: Hypertriglyceridaemia, low high density lipoprotein (HDL) cholesterol level and reduced LDL particle size are the major features of uraemic dyslipidaemia. They are also found in the Insulin Resistance Syndrome. AIM: To examine alterations in HDL composition in patients on chronic dialysis and their relationship with insulin resistance. METHODS: HDL particle size was determined in 33 patients on chronic haemodialysis (HD), 27 on chronic ambulatory peritoneal dialysis (CAPD) and 32 control non-diabetic subjects (C) without renal disease by non-denaturing 3-30% polyacrylamide gradient gel electrophoresis. A weighted HDL particle size score was calculated taking into account both HDL particle size and percentage total HDL protein concentration of each HDL band of the individual. Lipid and apolipoliprotein concentrations were determined in HDL2 and HDL3 particles obtained by sequential ultracentrifugation. In a subset of 24 control subjects and 22 subjects on HD, insulin sensitivity was also determined by an intravenous glucose tolerance test (IVGTT). RESULTS: HDL particles were found to be more triglyceride enriched and apoAI depleted in subjects on HD even though plasma triglyceride level was highest in patients on CAPD. Five subpopulations of HDL particles were identified by gradient gel electrophoresis in all subjects combined. In the subgroup of subjects who underwent IVGTT, the weighted HDL particle size score correlated positively with HDL cholesterol level (r = 0.6, p < 0.0005), LDL particle size (r = 0.47, p < 0.001), and insulin sensitivity (r = 0.48, p < 0.001), and negatively with plasma triglyceride level (r = 0.37, p < 0.01). CONCLUSIONS: We conclude that even though HDL cholesterol is reduced to a similar level in subjects on both forms of dialysis for end stage renal failure, abnormalities of HDL composition are more marked in subjects on HD. Reduction in HDL particle size is linked with insulin resistance and accompanies reduction in LDL particle size and hypertriglyceridaemia. PMID- 9227813 TI - Socio-economic disadvantage, quality of medical care and admission for acute severe asthma. AB - BACKGROUND: In asthma, socio-economic and health care factors may operate by a number of mechanisms to influence asthma morbidity and mortality. AIM: To determine the quality of medical care including the patient perception of the doctor-patient relationship, and the level of socio-economic disadvantage in patients admitted to hospital with acute severe asthma. METHODS: One hundred and thirty-eight patients (15-50 years) admitted to hospital (general ward or intensive care unit) with acute asthma were prospectively assessed using a number of previously validated instruments. RESULTS: The initial subjects had severe asthma on admission (pH = 7.3 +/- 0.2, PaCO2 = 7.1 +/- 5.0 kPa, n = 90) but short hospital stay (3.7 +/- 2.6 days). Although having high morbidity (40% had hospital admission in the last year and 60% had moderate/severe interference with sleep and/or ability to exercise), they had indicators of good ongoing medical care (96% had a regular GP, 80% were prescribed inhaled steroids, 84% had a peak flow meter, GP measured peak flow routinely in 80%, 52% had a written crisis plan and 44% had a supply of steroids at home). However, they were severely economically disadvantaged (53% had experienced financial difficulties in the last year, and for 35% of households the only income was a social security benefit). In the last year 39% had delayed or put off GP visit because of cost. Management of the index attack was compromised by concern about medical costs in 16% and time off work in 20%. CONCLUSION: Patients admitted to hospital with acute asthma have evidence of good quality on-going medical care, but are economically disadvantaged. If issues such as financial barriers to health care are not acknowledged and addressed, the health care services for asthmatics will not be effectively utilised and the current reductions in morbidity and mortality may not be maintained. PMID- 9227814 TI - Adult acute leukaemia--a retrospective study of 66 consecutive patients. AB - BACKGROUND: Advances in therapy and supportive care have improved the outlook for many patients with acute leukaemia, however elderly patients have increased treatment-related toxicity and shorter survival. Most clinical trials have selected patient populations with young median ages and it is therefore difficult to apply results to the general population where the majority of patients presenting with acute leukaemia are over 60 years. There is little information in the literature guiding appropriate treatment of these patients. AIMS: To determine the relationship between age, treatment received, and outcome in patients presenting with acute leukaemia. METHODS: A retrospective analysis was performed on all patients presenting to Prince Henry's Hospital and Monash Medical Centre with acute myeloid leukaemia (AML) or acute lymphocytic leukaemia (ALL) during a five year period. RESULTS: Sixty-six patients (51-AML, 15-ALL) presented with acute leukaemia between March 1989 and April 1994. Median patient age was 63 years; 32% of patients received supportive therapy only; 86% of patients with ALL and 58% of patients with AML commencing remission-induction chemotherapy entered a complete remission. Median survival for patients receiving only supportive therapy was three months. Median survival in patients under 55 years was almost twice as long as patients over 55 years receiving similar treatment (AML-eight vs four months p = 0.023; ALL-3.5 vs seven months p = 0.029). CONCLUSIONS: Median survival for acute leukaemia is inversely proportional to age. Applying results from selected series to elderly patients with acute leukaemia is inappropriate. PMID- 9227815 TI - The use of pamidronate in hypertrophic pulmonary osteoarthropathy (HPOA). AB - BACKGROUND: Hypertrophic pulmonary osteoarthropathy (HPOA) secondary to bronchogenic carcinoma can be associated with severe, disabling pain which is not always responsive to conventional treatment. AIM: To report on the use of pamidronate to control resistant pain in HPOA in three cases. METHODS: A retrospective review of reported pain, chest X-ray and radionuclide bone scans was made. RESULTS: Pain relief was achieved in all three cases together with reduced radiolabel uptake in two cases. CONCLUSIONS: Pamidronate appears to be an effective therapy for HPOA. Further investigation is warranted. PMID- 9227816 TI - Factors influencing the outcome of donor marrow transplantation in adults from less than ideal donors: experience from two Australian centres. AB - BACKGROUND: This paper reports the results of 78 marrow transplants in two Australian hospitals between 1991 and 1996, using unrelated (n = 54) or mismatched related (n = 24) donors. Twenty-six patients received granulocyte macrophage colony stimulating factor (GM-CSF) post-transplant as part of a phase II study. Fifty-four patients (74%) had advanced disease. AIMS: To identify factors associated with a superior outcome post-transplant, to evaluate the effect of GM-CSF on engraftment and other transplant parameters, and to compare the overall results with those of published series. METHODS: Review of patient records, a Medline search of the relevant literature and appropriate statistical analysis. RESULTS: The probability of overall survival and event-free survival (EFS) at three years was 35 +/- 6% and 22 +/- 6% respectively. Pre-transplant factors significantly associated with an inferior EFS were advanced disease, poorer performance status and age > 30 years. The EFS in patients with standard risk disease was 51 +/- 13% versus 10 +/- 5% in patients with advanced disease, p < 0.0001. Severe acute graft-versus-host disease was also associated with a poorer outcome. Neutrophil engraftment was faster in patients who received GM-CSF but there was no difference in any other transplant parameters. CONCLUSIONS: These results are consistent with reported series elsewhere and suggest that an extended family or unrelated donor transplant should generally be limited to patients with a good performance status and early phase but otherwise incurable haematological disease. PMID- 9227817 TI - Experience with the use of continuous positive airway pressure (CPAP) therapy in the emergency management of acute severe cardiogenic pulmonary oedema. AB - BACKGROUND: Acute pulmonary oedema (APO) is a frequent cause of respiratory failure and a common reason for presentation to emergency departments (EDs). To date, no paper has been published on the application of continuous positive airway pressure (CPAP) therapy for a large broad-based patient group. AIM: To report our experience with the use of CPAP in severe APO oedema, with particular reference to safety, intubation rates and impact on EDs' resources. METHOD: A retrospective chart review was undertaken of 75 patients with acute severe pulmonary oedema who were treated with adjuvant CPAP in an urban teaching hospital ED. RESULTS: Three patients (4%) required subsequent endotracheal intubation and mechanical ventilation. The average duration of CPAP was 1.9 hours. Eighty nine per cent of patients experienced no adverse events while being treated with GPAP. Five patients failed to tolerate the tight fitting mask necessitating removal of CPAP, three patients experienced arrhythmias related to underlying cardiac disease and two patients experienced mild transient hypotension. Seventy one per cent of patients were discharged from the ED to general medical wards. The in-hospital mortality for patients treated with CPAP was 15%. CONCLUSION: This series has demonstrated that CPAP therapy delivered via a face mask for the treatment of acute severe APO is safe and effective when applied to a broad range of patients. We recommend the use of CPAP therapy for all suitable patients presenting in severe APO irrespective of age or underlying pulmonary disease. PMID- 9227818 TI - Multiple cycles of aggressive chemotherapy prior to autologous peripheral stem cell transplantation for lymphoma. AB - BACKGROUND: Based on in vitro evidence of synergy between certain chemotherapeutic agents and of improved efficacy of cytotoxics administered as continuous infusions we devised a novel chemotherapy regimen for use in patients with lymphoma. AIMS: To assess the efficacy and tolerability of this regimen alone and in combination with autologous stem cell transplantation (ASCT) in patients with relapsed or refractory lymphoma. METHODS: The regimen consisted of cytosine arabinoside and etoposide given as five day continuous infusions, bolus doses of methotrexate and cyclophosphamide and four days of oral dexamethasone (MADEC), together with prophylactic G-CSF. Fifty-one patients aged 17 to 65 years (median 43) were treated; 34 had intermediate or high grade non-Hodgkin's lymphoma and 17 had Hodgkin's disease. Patients with responsive disease received a second cycle. RESULTS: Eleven patients (22%) achieved a complete response and 19 patients (38%) achieved a partial response, eight of whom were gallium negative. Previous chemosensitivity was highly predictive for response; minimal response was seen in patients with primary refractory disease. The probability of survival and event-free survival at 40 months for the entire group is 46% (+/- 8% SEM) and 26 +/- 8% respectively. Thirty-one patients proceeded to ASCT; the probability of survival of the transplanted group is 65 +/- 11% at 40 months. The regimen caused significant myelosuppression. The majority of cycles were complicated by febrile neutropenia and the requirement for platelet and red cell transfusions. Non-haematological toxicity was predominantly mucositis and was generally mild. CONCLUSIONS: MADEC is a highly active regimen in patients with relapsed lymphoma. Patients responding to MADEC who proceed to ASCT have a good chance of achieving a durable remission. PMID- 9227819 TI - The calcium channel blocker controversy in 1997. PMID- 9227820 TI - Pulmonary alveolar microlithiasis: an alternative diagnosis to miliary tuberculosis. PMID- 9227821 TI - Cerebral artery gas embolism (CAGE) following fine needle aspiration biopsy of the lung. PMID- 9227822 TI - Marfan syndrome and dural ectasia: a common, yet little known association. PMID- 9227823 TI - Microfilarial pleural effusion associated with adenocarcinoma. PMID- 9227824 TI - Pneumococcal arthritis and monoclonal gammopathy of undetermined significance. PMID- 9227825 TI - Nitritoid reaction in a patient on ACE inhibitor and Myocrisin treatments. PMID- 9227826 TI - SeHCAT tests for determination of bile acid malabsorption. PMID- 9227827 TI - Barmah Forest virus causing anaemia? PMID- 9227828 TI - Lung function test findings in patients with chronic fatigue syndrome (CFS) PMID- 9227829 TI - Acute pyelonephritis in a child caused by Proteus penneri. PMID- 9227830 TI - Pancreatic tuberculosis (TB) in HIV infection. PMID- 9227831 TI - Markedly delayed puberty in a male. PMID- 9227832 TI - Re-use of electrode catheters labelled as single use. PMID- 9227833 TI - Insights into the role of the cholinergic component of the septohippocampal pathway: what have we learned from experimental lesion studies? PMID- 9227834 TI - Dual effect of DMPP on the resting release of noradrenaline from rat hippocampal slices. AB - The effect of the nicotinic receptor agonist dimethylphenylpiperazinium iodide (DMPP) on the resting release of [3H]noradrenaline from superfused hippocampal slices was studied in rat. Continuous administration of DMPP at a concentration range of 1-100 microM increased the [3H]noradrenaline release in a dose-dependent manner. The response to DMPP was characterized by an immediate steep increase (peak response) followed by a sudden decline to a lower level that was constant with time (tall response) and still was significantly higher than the spontaneous release. Further analysis revealed that the release of noradrenaline in response to DMPP consists of two components. While nicotinic receptor antagonists (mecamylamine 10 microM, pancuronium 10 microM, pipecuronium 10 microM), the nonselective Ca-antagonist Cd2+ (125 microM) and tetrodotoxin (TTX, 1 microM) completely abolished the peak response (phase I), they had no effect on the tall response (phase II). Ca(2+)-free medium containing 1 mM EGTA also blocked phase I but in contrast with other drugs enhanced phase II. The release during phase I is subject to presynaptic feedback modulation, since the alpha 2-adrenoceptor agonist xylazine (3 microM) inhibited the DMPP-evoked stimulation of [3H]noradrenaline release, that inhibition was antagonized by a selective alpha 2 adrenoceptor antagonist, (+/-)-[7,8-(methylenedioxy)-14-alpha-hydroxyalloberbane hydrochloride [(+/-)-CH-38083] (2 microM). (+/-)-CH-38083 (2 microM) alone significantly enhanced the DMPP-evoked increase of [3H]noradrenaline release. Phase II was not effected by alpha 2-adrenergic drugs. Whereas the noradrenaline uptake blockers despramine (DMI, 1-10 microM), nisoxetine (1-10 microM), and nomifensine (10 microM) inhibited both phases, nomifensine at a concentration of 1 microM selectively blocked only phase II. Our data indicate that DMPP has a dual effect on the hippocampal noradrenaline release: phase I is a transient, nicotinic receptor-mediated exocytotic release, and phase II is a maintained, carrier-mediated process. PMID- 9227835 TI - Effects of noxious hindpaw immersion on evoked and spontaneous firing of contralateral convergent dorsal horn neurons in both intact and spinalized rats. AB - The effects of immersing a hindpaw in 60 degrees C water for 15 s on spontaneous and pinch-evoked activity recorded from 77 convergent neurons in the contralateral lumbar enlargement were examined. Cells were recorded from intact rats anesthetized with either ketamine or pentobarbital, or in rats that were decerebrated-intact or decerebrated-spinalized to determine if the heterotopic modulation was altered under these different conditions. In 47 neurons with background firing, immersion of the contralateral hindpaw inhibited the on-going activity in 6 of 19 cells recorded from ketamine anesthetized rats, 4 of 12 from pentobarbital anesthetized rats, 7 of 12 from decerebrate-spinalized rats, and 0 of 4 from decerebrate-intact rats. However, this inhibition was variable, both between and within neurons, because not every convergent neuron was inhibited, and there were trials in which immersion had no effect on the on-going activity of a neuron that had been previously inhibited by identical stimulation. In all 77 neurons, across all groups, discharges evoked by pinching the ipsilateral hindpaw were unaffected by contralateral immersion, even in cells where on-going activity was inhibited. Immersion of a hindpaw inhibited on-going, but not evoked, activity of contralateral convergent neurons. This inhibition was observed in rats under varied anesthetic conditions and in spinalized and intact rats. However, not all convergent neurons were inhibited by the remote stimulation even in intact rats, an observation inconsistent with the DNIC model of supraspinally mediated heterotopic modulation. This indicates that the effect of remote noxious stimulation in intact animals is not, under every experimental condition, a widespread inhibition of all convergent neurons. PMID- 9227836 TI - The activation of calcium/calmodulin-dependent protein kinase II after glutamate or potassium stimulation in hippocampal slices. AB - Two forms of long-term potentiation (LTP), N-methyl-D-aspartate receptor (NMDAR) dependent and non-NMDAR dependent, have been reported in hippocampal CA1 and CA3, respectively. The present study examined the activation of CaM-kinase II (calcium/calmodulin-dependent protein kinase II) in CA1 and CA3 areas after glutamate or potassium stimulation. Rat hippocampal slices were preincubated with one of the drugs (EGTA, DL-APV, CNQX, AP3, nitrendipine, KN-62, staurosporin, and H-89) before they were stimulated with either glutamate/glycine (100 microM/1 microM) or KCl (60 mM). Hippocampal CA1 area and CA3 area were then dissected and CaM-kinase II activities were assayed in vitro. Glutamate and KCl stimulations enhanced the percentage of Ca(2+)-independent CaM-kinase II activity in CA1 area. This enhancement was suppressed by EGTA, DL-APV, CNQX, or KN-62, suggesting that the neuronal stimulation effect in CA1 area was mediating through NMDA receptors. Conversely, there was no significant enhancement of CaM-kinase II activity in the CA3 area after glutamate or KCl stimulation. Nevertheless, the percentage of calcium-independent CaM-kinase II activity in the CA3 area was suppressed by EGTA, nitrendipine, KN-62, staurosporin, or H-89, indicating that the activity of CaM-kinase II in the CA3 area was independent of NMDA receptor activation. PMID- 9227837 TI - Effect of ammonia on motor function in adult rats. AB - Changes in motor function were assessed in male rats after injecting graded doses (100, 200, 400, and 800 mg/kg, IP) of ammonium chloride and ammonium acetate. The effects were correlated with the concentrations of ammonia and glucose in the brain and blood. Spontaneous motor activity and motor coordination were inhibited after injecting 100 and 200 mg/kg, whereas with 400 and 800 mg/kg the animals exhibited convulsive movements. A dose-dependent increase was found in the concentrations of ammonia and glucose in both blood and brain. These were restored, 25 min after treatment, to control levels in the blood and not in the brain. A correlation was found between the time courses of inhibitory motor events and a rise in brain ammonia levels. Convulsant action of ammonium salts was accompanied by a marked elevation of ammonia and glucose concentration in the brain. The findings suggest that detoxication of diffused ammonia is a rate limiting process in the brain and that ammonia, at toxic concentrations, decreases glucose utilization in the brain, resulting in an inhibition of motor function. A very high concentration of ammonia in the brain, although inhibiting glucose utilization, produces clonic convulsions probably by activating directly the motor neurons. PMID- 9227838 TI - Extensive cytotoxic lesions involving both the rhinal cortices and area TE impair recognition but spare spatial alternation in the rat. AB - Rats with cytotoxic lesions of the perirhinal, postrhinal, and TE cortices (Rh+TE, n = 7) were compared with surgical control animals (n = 7) on a series of spontaneous object recognition tests. The Rh+TE group was associated with a failure to select the novel object. This recognition deficit contrasted with the apparently normal ability of the same animals to learn and perform a spatial working memory test (T-maze alternation). The animals were also tested on the acquisition of an automated visual discrimination task in which the stimuli were presented on a visual display unit (VDU) equipped with a touch screen. The animals with Rh+TE lesions showed only a borderline deficit on this task. These findings are consistent with other evidence implicating the rhinal region in recognition memory. More importantly, they also provide a dissociation between spatial working memory and object recognition and, hence, show that extensive rhinal lesions are not sufficient to disconnect the hippocampus functionally. PMID- 9227839 TI - Altered behaviour in hamsters conceived and born in hypergravity. AB - We studied vestibular function in 37 hamsters (1 month old) conceived and born in either hypergravity (n = 21) or normal gravity (n = 16). Four groups were made: (1) HL group: 20 weeks in 2.5 G and 14 weeks in 1 G; (2) HS group: 4 weeks in 2.5 G and 30 weeks in 1 G; (3) CON group: 34 weeks in 1 G; and (4) ROT group: 4 weeks in 1 G, 16 weeks in rotation in 1 G, at the centre of the centrifuge and 14 weeks 1 G. When the hamsters were 4 weeks old, their locomotor activity, swimming ability, and air-righting was assessed. We found that HL and HS hamsters had no disturbances during locomotion in 1 G but their swimming ability was disturbed (swimming underwater, circling, and decreased speed of swimming). The HL hamsters showed less activity during 2.5 G and showed fewer correct air-rightings than the other groups. Differences between groups in swimming ability and the number of correct air-righting responses remained even after 3 months of normal gravity. Based on these findings, we suggest that the persistent behavioural disturbances are caused by the embryonal development of the hamsters in a hypergravity environment. Furthermore, hypergravity and rotation each have a different effect on behaviour. PMID- 9227841 TI - Effects of nicergoline on age-related decrements in radial maze performance and acetylcholine levels. AB - The effects of chronic oral administration of nicergoline (5.0 mg/kg; bid) on locomotor activity, eight-arm radial maze performance plus striatal, cortical, and hippocampal acetylcholine (ACh) levels were examined in young and aged Wistar rats. Chronic nicergoline administration did not modify either the locomotor activity or radial maze learning in young rats. Young rats learned the radial maze procedure rapidly and improved their performance throughout the successive training sessions. Radial maze performance in young rats was characterised by very few arm reentries. Aged rats were hypoactive and did not explore or enter the radial maze arms, and consequently performed poorly in the radial maze throughout the training sessions. Nicergoline treatment did not significantly modify locomotor activity in aged rats. Aged rats treated with nicergoline also performed poorly initially but improved with repeated training in the radial maze. This improvement was associated with an increasing number of arms being entered and very few arm reentries. Reduced acetylcholine (ACh) levels were also associated with age. Aged rats had significantly reduced levels of ACh in the straitum and cortex, but not the hippocampus as compared to young rats. Nicergoline treatment did not change ACh levels in young rats, but substantially restored the reduced ACh levels in aged rats. These results indicate that nicergoline is an effective cognitive enhancer in a learning model of age-related deficits and that these results may be related to changes in the cholinergic system. PMID- 9227840 TI - Is binding to nicotinic acetylcholine and dopamine receptors related to working memory in rats? AB - Nicotinic acetylcholine (ACh) and dopamine (DA) receptor activation has been found to be important for working memory. The regional distribution of these receptors in the brain has been well characterized. However, the relationship of the region-specific nicotinic ACh and DA binding density to memory performance has not been well assessed. In the current studies the relationship of receptor binding and memory function was examined. Receptor binding and memory performance were assessed in rats in three types of conditions: 1) chronic nicotine and mecamylamine vs. vehicle infusion; 2) lesions of the fimbria-fornix or medial basalocortical projection vs. sham lesions; and 3) 2-year-old aged rats vs. 3 month-old young adult rats. Nicotinic ACh receptors were labeled by [3H]N-methyl carbamylcholine ([3H]MCC), D1 receptors by [3H]SCH 23390, and D2 receptors by [125I]iodosulpiride. Working memory was assessed using the radial-arm maze and T maze delayed spatial alternation tasks. Chronic nicotine infusion substantially increased nicotinic receptor binding in a variety of brain areas and significantly improved working memory performance in the radial-arm maze. However, nicotinic receptor binding did not correlate well with memory performance. The nicotinic antagonist mecamylamine did not block nicotine-induced increased nicotinic binding, but it did block nicotine-induced memory improvement. Aged rats relative to young adults showed both a decrease in nicotinic binding and impaired memory performance. However, chronic effects of nicotine on nicotinic receptor binding and memory performance did not correlate in the aged rats. Nicotine also increased nicotinic receptor binding in the aged rats in brain areas except for the VTA, but did not improve memory performance. Lesions of the medial basalocortical projection or the fimbria-fornix did not cause significant changes in nicotinic binding in their target fields, but they did cause significant deficits in memory performance. Finally, there were no significant correlations of nicotinic binding in any brain region and memory performance. DA receptor binding was not altered by chronic nicotine or mecamylamine infusion, fimbria-fornix lesions, medial basalocortical lesions, or in aged rats. However, DA receptor binding did correlate with memory performance. There was a positive correlation of T-maze accuracy and D1 receptor binding in the frontal cortex and a negative correlation of T-maze accuracy and D1 receptor binding in the VTA and dentate gyrus. In contrast, a positive correlation was seen between radial-arm maze accuracy and D1 receptor binding in the VTA. Radial arm maze accuracy was positively correlated with D2 receptor binding in the striatum and dentate gyrus. There are significant relationships between the extent of DA receptor binding and working memory, but relationship between nicotinic ACh receptor binding density and memory is weak. PMID- 9227842 TI - Effect of posterior pituitary denervation (PPD) on prolactin (PRL) and alpha melanocyte-stimulating hormone (alpha-MSH) secretion of lactating rats. AB - Previous data have clearly suggested that the posterior pituitary (PP), consisting of neural lobe (NL) and intermediate lobe (IL), has a role in the control of anterior pituitary PRL secretion. However, basic aspects of this regulatory mechanism like (1), the role of an intact hypothalamic innervation of the PP as well as (2) the site of production of previously found PRL releasing substance(s) have not yet been characterized. Denervation of the PP (PPD) is an effective method for having a selective lesion of the innervation of PP, indeed, PPD results in a disappearance of neurosecretory materials from NL and tyrosine hydroxylase (TH) immunoreactivity from IL, leaving blood supply of all three lobes intact. Blood samples were taken from freely moving sham an PP-denervated lactating rats before and after 4-h separation from their pups and during the suckling stimulus. PPD blocks separation-induced depletion but only attenuates suckling induced release of PRL. Furthermore, it doubles plasma level of alpha MSH during the entire sampling period, which has been used as a marker for in vivo secretory activity of IL cells. Lack of the separation-induced depression in plasma PRL of PPD animals can be partially restored by normalizing the diabetes insipidus with treatment of a vasopressin analogue, 1-desamino-8-D-arginine vasopressin (dDAVP). In contrast, dDAVP, neither alone nor in combination with oxytocin (OXY), can change PPD-induced elevation of plasma alpha-MSH as well as attenuation of PRL response induced by suckling. It is concluded that: (1) contribution of the THDA system parallel to the confirmed role in the regulation of alpha-MSH seems to be crucial for the depletion of plasma PRL induced by separation but not for the elevation due to suckling stimulus, (2) intact hypothalamic innervations of both NL and IL, regulating water intake and the secretion of alpha-MSH, respectively, are necessary for normal secretory responses of AL during lactation, (3) as well as for the presence of PRF activity in PP, (4) which does not solely responsible for suckling-induced PRL release. Therefore, an interplay between several substances produced by NIL of the pituitary gland must have been responsible for the intact regulation of PRL secretion during lactation. PMID- 9227843 TI - Effect of intrathecal pretreatment with the neurokinin receptor antagonist CP 99994 on the expression of naloxone-precipitated morphine withdrawal symptoms. AB - In morphine-dependent rats pretreated with an intrathecal injection of saline (vehicle), intraarterial injection of 0.5 mg/kg of naloxone produced an immediate increase in blood pressure. Heart rate increased in most rats just after naloxone injection; however, the responses were transient, not lasting more than about 4 min after injection. Naloxone-precipitated behavioral changes were dominated by the appearance of body shakes and escape attempts that were strongly expressed during the first 10 min after naloxone. Pretreatment of morphine-dependent rats with an intrathecal injection of 100 nmol of the neurokinin-1 receptor antagonist CP-99994 significantly inhibited the magnitude and shortened the duration of the pressor response to naloxone. CP-99994 did ot reduce the expression of the associated withdrawal behaviors. Substance P significantly reversed the inhibitory effects of CP-99994 on the expression of the withdrawal-associated pressor response. Intrathecal pretreatment with CP-99994 also produced a dose dependent inhibition of the expression of the pressor response to local spinal (intrathecal) injection of naloxone (60 micrograms) in morphine dependent rats without significant alteration of the expression of withdrawal-associated behaviors. These results indicate that spinal neurokinin-1 receptors mediate some of the cardiovascular signs of morphine withdrawal and suggest the possibility of developing a novel class of antiopiate withdrawal agents. PMID- 9227844 TI - S-100-positive glial cells are involved in the regeneration of the visual pathway of teleosts. AB - Glial cells in the normal and regenerating visual pathways of Tinca tinca (Cyprinid, Teleost) were studied by labelling with anti-S-100 antibody. In normal fish, S-100-positive bipolar cells were found in the optic nerve, optic tract, and in the diencephalic visual pathways. After crushing the left optic nerve, the distribution and the number of S-100-immunoreactive cells were modified. In the injured nerve, 7 to 15 days after crushing no immunoreactive cell bodies were found in the crushed area, but a greater number of S-100-positive cells were found on both sides of the injured area. Sixty days after crushing, positive cells penetrating the crushed area were observed; the normal pattern was almost restored 200 days after crushing. In the diencephalon, 25 days after crushing, the number of S-100-positive cells increased remarkably and the most intense immunostaining of glial processes was observed 60 days after crushing. The density of S-100-labelled cells decreased after 4 months postcrushing. However, in the optic tectum no changes were observed. The increase of glial cells in the lesioned visual pathway suggests that they could play an important role in axonal regeneration after crushing. PMID- 9227845 TI - 24-hour rhythmicity of NADPH-diaphorase activity in the neuropil of rat visual cortex. AB - In the present study we examined the daytime-dependent alterations of nitric oxide synthase in the visual cortex of the rat. For this purpose, the activity of the nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d), an enzyme equivalent to nitric oxide synthase, was measured histochemically in rat visual cortex at 0600, 1200, 1800, and 2400 h using a photometric scanning method. Our results show day-time-dependent changes of the NADPH-d activity in the neuropil of the visual cortex. This was highest at 0600 h and decreased between 1200 h and 1800 h (unimodal profile of circadian activity). The number of NADPH-d-positive neuronal somata was not found to vary at the different time points. PMID- 9227846 TI - Regional differences of callosal connections in the granular zones of the primary somatosensory cortex in rats. AB - The primary somatosensory cortex (SI) in rats is cytoarchitectonically divided into three zones: the granular, peri-, and dysgranular zones. To examine callosal connections in the granular zone that bears representation of the body somatosensory map, the distribution of lectin-conjugated horse-radish peroxidase labeis was explored in the right SI after single or multiple injections into the granular zone of the left SI. After injections in the upper and lower law regions, many labeled cell bodies and dense terminal labeling were found in the regions homotopical to the injection sites. Both kinds of labels were densely seen in layers II-III and V, less densely in layer VI. Densely labeled terminals were also observed in layer I. In layer IV, many terminals and a few cell bodies were labeled in the septa, while labeling inside the barrels was sparse or absent. After injections in other regions, i.e., those representing the facial whiskers, fore- and hindlimbs, or trunk in the granular zone, labeled callosal cell bodies and terminals were sparse or absent, except in the septa of the posteromedial barrel subfield representing the facial whiskers. The results clearly show that the density of callosal connection in the granular zone differs in different subfields, and that at least the jaw regions in the granular zones of both hemispheres are directly interconnected, in contrast to the previous assumption that only the dysgranular zone mediates information transmission between the granular zones of both sides. PMID- 9227847 TI - Effect of morphine on proenkephalin gene expression in the rat brain. AB - Previous studies indicate that an acute injection of morphine does not effect the level of opioid peptides and their mRNA in the brain. However, due to the presence of a large pool of mRNA and possible opposing changes in turnover rate it is often difficult to visualize the transitory and relatively small alterations in gene transcription by examining mRNA level. Therefore, in situ hybridization with probes directed against intronic sequences to measure the primary transcript of proenkephalin (PPE) mRNA (heteronucleic RNA, hnRNA) in the rat brain following morphine administration was used in this study. The distribution of the hybridization signal of probes against both the A and B intron of the PPE gene were identical and coincide with the distribution PPE mRNA. Thus, to increase the sensitivity of this assay both probes were concurrently hybridized. Female and male Sprague-Dawley rats were gonadectomized and injected with morphine (10 mg/kg, SC). We detected no changes in PPE mRNA levels in the striatum, olfactory tubercle (OT) and n. accumbens core (NAC) at any time following morphine administration. However, from 0.5 h until 24 h following morphine injection, the levels of PPE hnRNA in NAC and OT but not in the dorsal striatum were significantly decreased. The level of c-fos mRNA was increased only the dorsal striatum following morphine injections. These data show that morphine administration can acutely change opioid peptide gene transcription. The observed decrease of PPE hnRNA levels for 24 h following a single morphine injection may indicate its importance for the development of acute and chronic dependence. However, the significance of these alterations in PPE gene transcription in term of the acute effect of morphine is not clear, because the steady-state level of mRNA was not changed. PMID- 9227848 TI - Vagal paraganglia bind biotinylated interleukin-1 receptor antagonist: a possible mechanism for immune-to-brain communication. AB - Interleukin-1 beta is a proinflammatory cytokine released by activated immune cells. In addition to orchestrating immune responses to infection, interleukin-1 beta is a key mediator of immune-to-brain communication. Interleukin-1 beta and endotoxin (which releases IL1 beta from immune cells) cause centrally mediated illness responses such as fever, aphagia, etc. These effects are blocked by intraperitoneal IL1 receptor antagonist (IL1ra), suggesting critical involvement of peripheral IL1 receptors. Centrally mediated illness responses are also blocked by vagotomy, suggesting that IL1 beta directly or indirectly activates vagal afferents. To test for IL1 beta binding whole vagus (abdominal, laryngeal, and thoracic) and sections of hepatic vagus and liver hilus were incubated with biotinylated IL1ra and processed for avidin-biotin complex (ABC) or avidin-FITC histochemistry. Glomus cells of vagal paraganglia were labeled in all regions of the vagus. Biotinylated IL1ra also labeled smooth muscle and endothelial cells of blood vessels and lymphoid tissues. No label was present in omission or competition controls. These data suggest that centrally mediated illness responses result from IL1 activation of vagal paraganglia. PMID- 9227849 TI - Premyofibrils in spreading adult cardiomyocytes in tissue culture: evidence for reexpression of the embryonic program for myofibrillogenesis in adult cells. AB - Do adult cardiomyocytes use the same pathways hypothesized for the formation of myofibrils in embryonic cardiomyocytes in tissue culture. [Rhee, et al., Cell Motil. Cytoskeleton 28:1-24, 1994]? Premyofibrils in embryonic cardiomyocytes are composed of short sarcomeric units of alpha-actinin (Z-bodies) and actin filaments held together by short nonmuscle myosin IIB filaments. Premyofibrils are believed to be transformed into nascent myofibrils by their capture of muscle specific myosin II filaments aligned in aperiodic arrays. Nascent myofibrils are thought to transform into mature myofibrils by the loss of nonmuscle myosin IIB, the fusion of the Z-bodies into Z-bands, and the periodic alignment of muscle myosin II filaments into A-bands. Freshly isolated cat and rat adult cardiomyocytes placed in tissue culture lack premyofibrils and nascent myofibrils. Adult cardiomyocytes spreading in culture reinitiate the synthesis of nonmuscle myosin IIB. Moreover, patterns similar to the proposed embryonic myofibrillar program first detected in spreading chick embryonic hearts were also detected in these spreading adult mammalian cardiomyocytes. The isolated adult cardiomyocytes begin to spread after 1 day in culture by sending out lamellipodia. When these cells are injected with fluorescently labeled alpha actinin, linear arrays of short spacings of beaded alpha-actinin bodies are detected in the spreading edges of the adult cardiomyocytes. These dense bodies (Z-bodies) stain positively for the same sarcomeric-specific isoform of alpha actinin that is in the Z-bands of mature sarcomeres. These linear arrays of alpha actinin-containing Z-bodies have other characteristics of premyofibrils and are detected only in the spreading regions of the cells. Thus, these premyofibrils at the edges of the spreading adult cardiomyocytes stain positively for nonmuscle myosin IIB but negatively for muscle-specific myosin II. Initially, no vinculin is associated with any parts of the premyofibrils in the spreading regions of the early spreading cardiomyocytes. However, later, vinculin is found to be associated with the ends of the premyofibrils. Fibers that stain solidly for muscle-specific myosin II (i.e., nascent myofibrils) are localized between the peripheral premyofibrils and the centrally positioned, mature myofibrils. It is suggested that the puzzling ability of cardiomyocytes in hypertrophic hearts to reinitiate the synthesis of fetal sarcomeric proteins may be related to the reinitiation of the embryonic premyofibril program for myofibrillogenesis. PMID- 9227851 TI - Capping of the barbed ends of actin filaments by a high-affinity profilin-actin complex. AB - Profilin, a ubiquitous 12 to 15-kDa protein, serves many functions, including sequestering monomeric actin, accelerating nucleotide exchange on actin monomers, decreasing the critical concentration of the barbed end of actin filaments, and promoting actin polymerization when barbed ends are free. Most previous studies have focused on profilin itself rather than its complex with actin. A high affinity profilin-actin complex (here called profilactin) can be isolated from a poly-(L)-proline (PLP) column by sequential elution with 3 M and 7 M urea. Profilactin inhibited the elongation rate of pyrenyl-G-actin from filament seeds in a concentration- and time-dependent manner. Much greater inhibition of elongation was observed with spectrin-F-actin than gelsolin-F-actin seeds, suggesting that the major effect of profilactin was due to capping the barbed ends of actin filaments. Its dissociation constant for binding to filament ends was 0.3 microM; the on- and off-rate constants were estimated to be 1.7 x 10(3) M 1 s-1 and 4.5 x 10(-4) s-1, respectively. Purified profilin (obtained by repetitive applications to a PLP column and assessed by silver-stained polyacylamide gels) did not slow the elongation rate of pyrenyl-G-actin from filament seeds. Capping protein could not be detected by Western blotting in the profilactin preparation, but low concentrations of gelsolin did contaminate our preparation. However, prolonged incubation with either calcium or EGTA did not affect capping activity, implying that contaminating gelsolin-actin complexes were not primarily responsible for the observed capping activity. Reapplication of the profilactin preparation to PLP-coupled Sepharose removed both profilin and actin and concurrently eliminated its capping activity. Profilactin that was reapplied to uncoupled Sepharose retained its capping activity. Phosphatidylinositol-4,5-bisphosphate (PIP2) was the most potent phosphoinositol in reducing the capping activity of profilactin. Dissociation of the tight profilactin complex may serve as a unique mechanism by which profilin helps regulate actin filament growth. PMID- 9227850 TI - Mitochondrial inheritance: cell cycle and actin cable dependence of polarized mitochondrial movements in Saccharomyces cerevisiae. AB - Asymmetric growth and division of budding yeast requires the vectorial transport of growth components and organelles from mother to daughter cells. Time lapse video microscopy and vital staining were used to study motility events which result in partitioning of mitochondria in dividing yeast. We identified four different stages in the mitochondrial inheritance cycle: (1) mitochondria align along the mother-bud axis prior to bud emergence in G1 phase, following polarization of the actin cytoskeleton; (2) during S phase, mitochondria undergo linear, continuous and polarized transfer from mother to bud; (3) during S and G2 phases, inherited mitochondria accumulate in the bud tip. This event occurs concomitant with accumulation of actin patches in this region; and (4) finally, during M phase prior to cytokinesis, mitochondria are released from the bud tip and redistribute throughout the bud. Previous studies showed that yeast mitochondria colocalize with actin cables and that isolated mitochondria contain actin binding and motor activities on their surface. We find that selective destabilization of actin cables in a strain lacking the tropomyosin 1 gene (TPM1) has no significant effect on the velocity of mitochondrial motor activity in vivo or in vitro. However, tpm1 delta mutants display abnormal mitochondrial distribution and morphology; loss of long distance, directional mitochondrial movement; and delayed transfer of mitochondria from the mother cell to the bud. Thus, cell cycle-linked mitochondrial motility patterns which lead to inheritance are strictly dependent on organized and properly oriented actin cables. PMID- 9227852 TI - Kinesin-driven microtubule motility in the presence of alkaline-earth metal ions: indication for a calcium ion-dependent motility. AB - We studied the effect of alkaline-earth metal ions on the kinesin-driven gliding of microtubules, using a narrow glass chamber enabling the exchange of buffer components without interrupting microscopic observation. Under standard conditions (0.5 mM Mg2+), microtubules were found to glide at a mean velocity of about 0.6 micron/s. Motility was widely ceased after removing Mg2+. Subsequent addition of Ca2+ restored motility (maximal mean gliding velocity measured: 0.26 micron/s at 2.5 mM Ca2+). Also in the presence of Sr2+ or Ba2+ a slow gliding could be observed (0.025 micron/s and 0.014 micron/s, respectively, at 0.5 mM). After removal of Ca2+, Sr2+, or Ba2+ and re-addition of Mg2+, the gliding velocities reached approximately the values determined under standard conditions. Motility was not changed when 0.5 mM Ca2+, Sr2+, or Ba2+ were applied together with Mg2+. Microtubule gliding stopped after substitution of 0.5 mM BeCl2 for Mg2+. When both BeCl2 and Mg2+ were present, the mean gliding velocity was reduced to 0.29 micron/s. In addition, many microtubules were released from the kinesin coated glass surface, indicating that the beryllium salt disorders the binding between kinesin and microtubules. Our results confirm that Mg2+ is the most suitable cofactor for kinesin driven microtubule motility. However, they also demonstrate that brain kinesin can generate motility when Ca2+ was substituted for Mg2+. PMID- 9227853 TI - Induction of temporary beating in paralyzed flagella of Chlamydomonas mutants by application of external force. AB - To help understand the mechanism by which the sliding movement of outer-doublet microtubules in cilia and flagella is converted into bending waves, we examined the effect of mechanical force imposed on the flagella of Chlamydomonas mutants lacking the central pair or multiple dyneins. These mutants were almost completely nonmotile under normal conditions. A bend was produced in a flagellum either by holding a cell with a micropipette and quickly moving it with a piezoelectric actuator; or by pushing a flagellum with a microneedle. After removal of the external force, mutants lacking the central pair (pf18 and pf19) displayed beating at irregular intervals of > 1 second for one to several cycles. Similarly, a double mutant (ida2ida4) lacking four species of inner-arm dynein displayed beating at intervals of > 0.1 second for up to 80 cycles. However, paralyzed flagella of double mutants that lack the outer dynein arm in addition to the central pair or the inner dynein arm did not show cyclical movements upon application of external force. These results indicate that the central pair and the inner dynein arm are important for both stable bend formation at the base and efficient bend propagation along the flagellar length. They also suggest that the outer dynein arm, and not the inner dynein arm, enables the flagellar axoneme to propagate bends independently of the central pair. We propose that the axoneme is equipped with two independent motor systems for oscillatory movements: an outer arm system controlled by the axonemal mechanical state independently of the central pair/radial spoke system, and an inner-arm system controlled by both the axonemal mechanical state and the central pair/radial spokes. PMID- 9227854 TI - Myosin VIIa, the product of the Usher 1B syndrome gene, is concentrated in the connecting cilia of photoreceptor cells. AB - Usher syndrome is the most common form of combined deafness and blindness. The gene that is defective in Usher syndrome 1B (USH1B) encodes for an unconventional myosin, myosin VIIa. To understand the cellular function of myosin VIIa and why defects in it lead to USH1B, it is essential to determine the precise cellular and subcellular localization of the protein. We investigated the distribution of myosin VIIa in human and rodent photoreceptor cells and retinal pigment epithelium (RPE), primarily by immunoelectron microscopy, using antibodies generated against two different domains of the protein. In both human and rodent retinae, myosin VIIa was detected in the apical processes of the RPE and in the cilium of rod and cone photoreceptor cells. Immunogold label was most concentrated in the connecting cilium. Here, myosin VIIa appeared to be distributed outside the ring of doublet microtubules near the ciliary plasma membrane. These observations indicate that a major role of myosin VIIa in the retina is in the photoreceptor cilium, perhaps in such a function as trafficking newly synthesized phototransductive membrane or maintaining the diffusion barrier between the inner and outer segments. Our results support the notion that defective ciliary function is the underlying cellular abnormality that leads to cellular degeneration in Usher syndrome. PMID- 9227855 TI - Actin in the parasite Toxoplasma gondii is encoded by a single copy gene, ACT1 and exists primarily in a globular form. AB - Actin is a highly conserved microfilament protein that plays an important role in the invasion of host cells by the protozoan parasite Toxoplasma gondii. We have characterized the ACT1 gene and localized the conventional isoform of actin that it encodes within T. gondii. The predicted amino acid sequence of ACT1 was most similar to two other parasite actins, Plasmodium falciparum Pfact-1 (93.1% identical) and Cryptosporidium parvum actin (88.1%): among vertebrate actins, ACT1 was most closely related to the mammalian beta and gamma (83%) actin isoforms. Actin-specific antibodies and fluorescently labeled DNAse I were used to localize actin in T. gondii tachyzoites by immunofluorescence and immunoelectron microscopy. Actin was detected beneath the parasite cell membrane and in clusters scattered within the cytosol of T. gondii tachyzoites. Actin filaments were not detected in detergent-solubilized parasites separated by high speed centrifugation, indicating that actin exists primarily in a globular form in T. gondii. PMID- 9227856 TI - Effects of intracellular pH on the mitotic apparatus and mitotic stage in the sand dollar egg. AB - The effect of change in intracellular pH (pHi) on mitosis was investigated in the sand dollar egg. The pHi in the fertilized egg of Scaphechinus mirabilis and Clypeaster japonicus, which was 7.34 and 7.31, respectively, changed by means of treating the egg at nuclear envelope breakdown with sea water containing acetate and/or ammonia at various values of pH. The mitotic apparatus at pHi 6.70 became larger than that of normal fertilized eggs; that is, the mitotic spindle had the maximal size, especially in length at pHi 6.70. The spindle length linearly decreased when pHi increased from 6.70 to 7.84. By polarization microscopy, the increase in birefringence retardation was detected at slightly acidic pHi, suggesting that the increase in size of the spindle is caused by the increase in the amount of microtubules in the spindle. At pHi 6.30, the organization of the mitotic apparatus was inhibited. Furthermore, slightly acidic pHi caused cleavage retardation or inhibition. By counting the number of the eggs at various mitotic stages with time after treating them with the media, it is found that metaphase was persistent and most of the S. mirabilis eggs were arrested at metaphase under the condition of pHi 6.70. It is concluded that at slightly acidic pH, the microtubules in the spindle are stabilized and more microtubules assembled than those in the normal eggs. PMID- 9227857 TI - Laminin-5 and modulation of keratin cytoskeleton arrangement in FG pancreatic carcinoma cells: involvement of IFAP300 and evidence that laminin-5/cell interactions correlate with a dephosphorylation of alpha 6A integrin. AB - Under normal culture conditions, epithelial cells of the FG line, derived from a pancreatic tumor, characteristically grow in mounds and fail to flatten efficiently onto their substrate. In such cells, keratin intermediate filaments (IFs) are concentrated in the perinuclear region. Furthermore, the IF associated protein, IFAP300, primarily localizes along these keratin bundles. Additionally, alpha 6 beta 4 integrin heterodimers localize in streaks or spots towards the edges of cells while alpha 3 beta 1 integrin is predominantly at cell-cell surfaces. Neither show any obvious interaction with IF. Remarkably, upon plating FG cells into medium containing soluble rat laminin-5, FG cells rapidly adhere and spread onto their substrate. Moreover, FG cells "capture" rat laminin-5 and place it basally in circles or arcs at areas of cell-substrate interaction. Double label immunofluorescence microscopy reveals colocalization of IFAP300 as well as alpha 6 beta 4 and alpha 3 beta 1 integrin with the polarized laminin-5. Concomitantly, alpha 6 integrin undergoes dephosphorylation on serine residue 1041. Laminin-5-induced rapid adhesion can be blocked by antibodies against the alpha 3 integrin subunit. In contrast, while alpha 6 integrin antibodies do not block laminin-5-induced rapid adhesion, they prevent FG cells from assuming an epithelial-like morphology. Keratin IF bundles associate with IFAP300-alpha 6 beta 4/alpha 3 beta 1 integrin complexes along the cell-substratum-attached surface of FG cells coincubated in laminin-5-containing medium. Coprecipitation results suggest that in these complexes, IFAP300 may associate with the alpha 6 beta 4 integrin heterodimer. Based on our results and published evidence that IFAP300 binds keratin in vitro [Skalli et al., 1994; J. Cell Biol. 125:159-170], we propose that laminin-5/FG cell interaction results in a novel integrin dephosphorylation event, which subsequently induces IFAP300 association with alpha 6 beta 4 integrin. IFAP300 then mediates the interaction of IFs with the cell surface via the alpha 6 beta 4 integrin heterodimer. PMID- 9227859 TI - Extrapulmonary pneumocystosis. AB - Extrapulmonary pneumocystosis is an exceedingly rare complication of Pneumocystis carinii pneumonia (PCP). Prior to the advent of the human immunodeficiency virus type 1 (HIV-1) epidemic, only 16 cases of extrapulmonary pneumocystosis in individuals who were immunocompromised by a variety of underlying diseases had been reported. Since the beginning of the HIV-1 and related PCP epidemic, at least 90 cases of extrapulmonary pneumocystosis have been reported. This review briefly presents a history of the discovery of P. carinii and its recognition as a human pathogen, the controversy regarding its taxonomy, and the epidemiology of this organism. A more detailed analysis of the incidence of extrapulmonary pneumocystosis in HIV-1-infected individuals and its occurrence despite widespread prophylaxis for PCP with either aerosolized pentamidine or systemic dapsone-trimethoprim is presented. The clinical features of published cases of extrapulmonary pneumocystosis in non-HIV-1-infected individuals are summarized and contrasted with those in HIV-1 infected individuals. The diagnosis of extrapulmonary pneumocystosis is discussed, and because clinical microbiologists and pathologists are the key individuals in establishing the diagnosis, the characteristic microscopic morphology of P. carinii as its appears when stained with a variety of stains is presented and reviewed. The review concludes with a brief discussion of treatments for extrapulmonary pneumocystosis. PMID- 9227862 TI - Toxin production by Campylobacter spp. AB - Of all the virulence factors that were proposed for Campylobacter jejuni and related species to cause disease in humans, the discovery of toxin production was the most promising but led to a rather confusing and even disappointing stream of data. The discussion of whether proteinaceous exotoxins are relevant in disease remains open. One important reason for this lack of consensus is the anecdotal nature of the literature reports. To provide a basis for an unbiased opinion, this review compiles all described exotoxins, compares their reported properties, and provides a summary of animal model studies and clinical data. The toxins are divided into enterotoxins and cytotoxins and are sorted according to their biochemical properties. Since many Campylobacter toxins have been compared with toxins of other species, some key examples of the latter are also discussed. Future directions of toxin research that appear promising are defined. PMID- 9227861 TI - Update on detection of bacteremia and fungemia. AB - The presence of microorganisms in a patient's blood is a critical determinant of the severity of the patient's illness. Equally important, the laboratory isolation and identification of a microorganism present in blood determine the etiologic agent of infection, especially when the site of infection is localized and difficult to access. This review addresses the pathophysiology and clinical characteristics of bacteremia, fungemia, and sepsis; diagnostic strategies and critical factors in the detection of positive blood cultures; characteristics of manual and instrument approaches to bacteremia detection; approaches for isolating specific microorganisms associated with positive blood cultures; and rapid methods for the identification of microorganisms in blood cultures. PMID- 9227858 TI - Yeast killer systems. AB - The killer phenomenon in yeasts has been revealed to be a multicentric model for molecular biologists, virologists, phytopathologists, epidemiologists, industrial and medical microbiologists, mycologists, and pharmacologists. The surprisingly widespread occurrence of the killer phenomenon among taxonomically unrelated microorganisms, including prokaryotic and eukaryotic pathogens, has engendered a new interest in its biological significance as well as its theoretical and practical applications. The search for therapeutic opportunities by using yeast killer systems has conceptually opened new avenues for the prevention and control of life-threatening fungal diseases through the idiotypic network that is apparently exploited by the immune system in the course of natural infections. In this review, the biology, ecology, epidemiology, therapeutics, serology, and idiotypy of yeast killer systems are discussed. PMID- 9227860 TI - Experimental investigation of herpes simplex virus latency. AB - The clinical manifestations of herpes simplex virus infection generally involve a mild and localized primary infection followed by asymptomatic (latent) infection interrupted sporadically by periods of recrudescence (reactivation) where virus replication and associated cytopathologic findings are manifest at the site of initial infection. During the latent phase of infection, viral genomes, but not infectious virus itself, can be detected in sensory and autonomic neurons. The process of latent infection and reactivation has been subject to continuing investigation in animal models and, more recently, in cultured cells. The initiation and maintenance of latent infection in neurons are apparently passive phenomena in that no virus gene products need be expressed or are required. Despite this, a single latency-associated transcript (LAT) encoded by DNA encompassing about 6% of the viral genome is expressed during latent infection in a minority of neurons containing viral DNA. This transcript is spliced, and the intron derived from this splicing is stably maintained in the nucleus of neurons expressing it. Reactivation, which can be induced by stress and assayed in several animal models, is facilitated by the expression of LAT. Although the mechanism of action of LAT-mediated facilitation of reactivation is not clear, all available evidence argues against its involving the expression of a protein. Rather, the most consistent models of action involve LAT expression playing a cis acting role in a very early stage of the reactivation process. PMID- 9227867 TI - SPECT and subtraction imaging of an ectopic parathyroid adenoma. AB - Primary hyperparathyroidism is a disease best managed surgically; resection of the lesion(s) is curative. Although the routine use of localizing procedures is controversial before surgery, it is generally agreed that these techniques are useful in cases of reexploration. The authors have recently studied a patient with long-standing hyperparathyroidism, in whom initial surgery was unsuccessful. Although planar imaging with Tc-99m sestamibi is usually sufficient for identification and localization of the lesion, both SPECT and subtraction imaging provided additional information that contributed to the success of the subsequent surgery. Single-proton emission computed tomography provided critical information about the position of the ectopic parathyroid adenoma in relation to the right submandibular salivary gland, whereas subtraction imaging confirmed that the focus was indeed a parathyroid lesion, and not merely an anatomic variant of a normal salivary gland. PMID- 9227864 TI - Nasal carriage of Staphylococcus aureus: epidemiology, underlying mechanisms, and associated risks. AB - Staphylococcus aureus has long been recognized as an important pathogen in human disease. Due to an increasing number of infections caused by methicillin resistant S. aureus (MRSA) strains, therapy has become problematic. Therefore, prevention of staphylococcal infections has become more important. Carriage of S. aureus appears to play a key role in the epidemiology and pathogenesis of infection. The ecological niches of S. aureus are the anterior nares. In healthy subjects, over time, three patterns of carriage can be distinguished: about 20% of people are persistent carriers, 60% are intermittent carriers, and approximately 20% almost never carry S. aureus. The molecular basis of the carrier state remains to be elucidated. In patients who repeatedly puncture the skin (e.g., hemodialysis or continuous ambulatory peritoneal dialysis [CAPD] patients and intravenous drug addicts) and patients with human immunodeficiency virus (HIV) infection, increased carriage rates are found. Carriage has been identified as an important risk factor for infection in patients undergoing surgery, those on hemodialysis or CAPD, those with HIV infection and AIDS, those with intravascular devices, and those colonized with MRSA. Elimination of carriage has been found to reduce the infection rates in surgical patients and those on hemodialysis and CAPD. Elimination of carriage appears to be an attractive preventive strategy in patients at risk. Further studies are needed to optimize this strategy and to define the groups at risk. PMID- 9227866 TI - Hyperthyroidism with or without pyramidal lobe Graves' disease or disseminated autonomously functioning thyroid tissue? AB - I-123 thyroid scintigrams performed in 349 patients were evaluated with a focus on specific thyroid gland vestiges, namely a pyramidal lobe or a thyroglossal duct. The detection of these vestiges in patients with hyperthyroidiam is indicative of autoimmune hyperthyroidism. In Graves' disease, stimulating thyrotropin (TSH) receptor antibodies cause a significantly more frequent appearance of vestiges of the thyroglossal tract. In contrast, disseminated autonomously functioning thyroid nodules rarely show a pyramidal lobe. The frequency of pyramidal lobe visualization in patients with Graves' disease differed significantly from the frequency in patients with multifocal or disseminated autonomously functioning nodules. In euthyroidism patients, the vestiges may be indicative of the diagnosis of iodine deficiency with or without latent primary hypothyroidism. In thyroid scintigraphy, the pyramidal lobe and the thyroglossal duct can be visualized more easily using I-123 instead of Tc-99m sodium pertechnetate. PMID- 9227868 TI - Intense muscle uptake of Tc-99m MDP and Ga-67 citrate in massive rhabdomyolysis. AB - A 6-year-old boy had an Addisonian crisis with hyponatremic shock and required resuscitation after cardiac arrest. Severe renal failure necessitated continuous dialysis. Rhabdomyolysis was identified because of myoglobinuria and gross elevation of creatinine kinase. In the course of his treatment, bone infection was suspected. A bone scan with Tc-99m MDP showed intense uptake in numerous major muscle groups throughout the body in keeping with massive rhabdomyolysis. Similar gross muscle uptake of Ga-67 citrate was less expected. In an additional bone scan obtained 15 days later, the MDP uptake showed less intense accumulation. PMID- 9227871 TI - Resting Tl-201 scintigraphy in the evaluation of myocardial sarcoidosis. AB - In patients with sarcoidosis, myocardial involvement is common and may be fatal. With extensive disease, the primary manifestations may include conduction abnormalities and arrhythmias, which may lead to sudden death. Myocardial perfusion scintigraphy may be the most accurate method to assess extent of myocardial involvement and response to therapy. PMID- 9227870 TI - Gallium scanning in the 'new' tuberculosis. AB - PURPOSE: To evaluate the findings and usefulness of Ga-67 scanning in recent cases of tuberculosis (TB). MATERIALS AND METHODS: The authors reviewed chest x ray films and Ga-67 citrate scans of 52 patients with culture-confirmed infection caused by Mycobacterium tuberculosis treated after 1988. RESULTS: Ga-67 scans were positive in every case, delineating upper lobe lung lesions in 6 patients, diffuse involvement or lower lobe disease in 34 patients, and intrathoracic adenopathy in 15 patients. Pulmonary parenchymal lesions were not detected on x ray films in 3 patients, and nodal lesions were not apparent in 3 patients. In addition, in 6 patients cervical adenopathy was detected by Ga-67 scintigraphy; 4 underwent biopsy with culture confirmation. CONCLUSIONS: Ga-67 scanning is more sensitive than routine chest radiography for detection of both TB parenchymal lung involvement and adenopathy. Ga-67 imaging facilitates the choice of biopsy sites by identifying accessible peripheral nodes. Typical patterns in recent cases of TB differ significantly from the upper lobe predilection of classical TB. PMID- 9227869 TI - Incidental appendix carcinoid. Value of somatostatin receptor imaging. AB - PURPOSE: Somatostatin receptor scintigraphy is used to diagnose carcinoid of the gastrointestinal tract. Its sensitivity ranges from approximately 75%-100%. Therefore, it was hypothesized that it can be used in the postsurgical follow-up to detect residual carcinoid, recurrence, and metastatic disease. RESULTS: This article is concerned with the findings of somatostatin receptor imaging performed on a 12-year-old girl 8 weeks after appendectomy. Histologic examination showed an incidental appendix carcinoid. Somatostatin receptor scintigraphy performed for detection of lymph node metastatic spread of the carcinoid showed focal tracer accumulation at the former operative site; subsequently, a right hemicolectomy was performed. However, histologic examination of the surgical tissue showed no evidence for carcinoid. CONCLUSION: It is concluded that there are some potential pitfalls for somatostatin receptor imaging at least soon after surgery. Therefore, it should not be used to aid in reoperation. PMID- 9227873 TI - Bone scan findings in Kienbock's disease. A case report with atypical findings and literature review. AB - The authors present an 18-year-old man who had a 5-month history of a painful left wrist. Despite the prolonged history, discrete photopenia on the blood-pool phase and photopenia relative to the remainder of the ipsilateral carpus on the delayed phase of a bone scan in the region of the lunate was shown. When Kienbock's disease is seen in its late phase, the bone scan findings may be atypical in that they may not show the usual three-phase bone scintigraphic evidence of bone remodeling expected in delayed diagnosis avascular necrosis. A review of the previous literature is presented. PMID- 9227863 TI - Antifungal prophylaxis during neutropenia and immunodeficiency. AB - Fungal infections represent a major source of morbidity and mortality in patients with almost all types of immunodeficiencies. These infections may be nosocomial (aspergillosis) or community acquired (cryptococcosis), or both (candidiasis). Endemic mycoses such as histoplasmosis, coccidioidomycosis, and penicilliosis may infect many immunocompromised hosts in some geographic areas and thereby create major public health problems. With the wide availability of oral azoles, antifungal prophylactic strategies have been extensively developed. However, only a few well-designed studies involving strict criteria have been performed, mostly in patients with hematological malignancies or AIDS. In these situations, the best dose and duration of administration of the antifungal drug often remain to be determined. In high-risk neutropenic or bone marrow transplant patients, fluconazole is effective for the prevention of superficial and/or systemic candidal infections but is not always able to prolong overall survival and potentially selects less susceptible or resistant Candida spp. Primary prophylaxis against aspergillosis remains investigative. At present, no standard general recommendation for primary antifungal prophylaxis can be proposed for AIDS patients or transplant recipients. However, for persistently immunocompromised patients who previously experienced a noncandidal systemic fungal infection, prolonged suppressive antifungal therapy is often indicated to prevent a relapse. Better strategies for controlling immune deficiencies should also help to avoid some potentially life-threatening deep mycoses. When prescribing antifungal prophylaxis, physicians should be aware of the potential emergence of resistant strains, drug-drug interactions, and the cost. Well designed, randomized, multicenter clinical trials in high-risk immunocompromised hosts are urgently needed to better define how to prevent severe invasive mycoses. PMID- 9227874 TI - Unilateral lung hypoperfusion with normal ventilation on pulmonary scintigraphy caused by pulmonary artery sling. PMID- 9227872 TI - Abnormal Tl-201 myocardial scintigraphy associated with administration of sodium aurothiomalate. AB - A patient with strikingly reduced Tl-201 myocardial retention after intramuscular administration of gold therapy (sodium aurothiomalate) is described. Following a routine Bruce protocol exercise test, postexercise scintigraphy was normal; an abnormal scan of the thorax was obtained on delayed imaging at 3 hours. This scan showed no demonstrable myocardial retention of tracer. In between the two acquisitions, the patient received 50 mg intramuscular sodium aurothiomalate for rheumatoid arthritis. A repeat scan obtained 3 days later showed a normal myocardial scintigram. It is recommended that patients receiving intramuscular gold therapy should not undergo such treatment close to the timing of Tl-201 myocardial scans. PMID- 9227875 TI - Radionuclide inferior venocavagraphy in Budd-Chiari syndrome before and after angioplasty. PMID- 9227876 TI - Intense accumulation of Tc-99m tetrofosmin in chronic pneumonia. PMID- 9227877 TI - 'False-positive' uptake of FDG in a hepatic adenoma. PMID- 9227878 TI - Autonomous thyroid adenoma, papillary thyroid carcinoma, and ectopic parathyroid adenoma in a patient with primary hyperparathyroidism and a nontoxic multinodular goiter. PMID- 9227880 TI - Ga-67 scintigraphy in a patient with infected vascular synthetic graft. PMID- 9227881 TI - Megacolon demonstrated on colonic transit imaging. PMID- 9227879 TI - Ureteroenteric fistula shown only on bone imaging. PMID- 9227882 TI - Scrotal cellulitis simulating testicular infarction by Tc-99m pertechnetate imaging. PMID- 9227883 TI - Medullary cavity stenosis of the metacarpals. Scintigraphic appearance. PMID- 9227884 TI - Redistribution. The sign of the successful testicular detorsion. PMID- 9227885 TI - Golfer's rib stress fracture (Duffer's fracture). Scintigraphic appearance. PMID- 9227865 TI - Human herpesvirus 6. AB - Human herpesvirus 6 variant A (HHV-6A) and human herpesvirus 6 variant B (HHV-6B) are two closely related yet distinct viruses. These visuses belong to the Roseolovirus genus of the betaherpesvirus subfamily; they are most closely related to human herpesvirus 7 and then to human cytomegalovirus. Over 95% of people older than 2 years of age are seropositive for either or both HHV-6 variants, and current serologic methods are incapable of discriminating infection with one variant from infection with the other. HHV-6A has not been etiologically linked to any human disease, but such an association will probably be found soon. HHV-6B is the etiologic agent of the common childhood illness exanthem subitum (roseola infantum or sixth disease) and related febrile illnesses. These viruses are frequently active and associated with illness in immunocompromised patients and may play a role in the etiology of Hodgkin's disease and other malignancies. HHV-6 is a commensal inhabitant of brains; various neurologic manifestations, including convulsions and encephalitis, can occur during primary HHV-6 infection or in immunocompromised patients. HHV-6 and distribution in the central nervous system are altered in patients with multiple sclerosis; the significance of this is under investigation. PMID- 9227886 TI - Liver and lung metastases of high-grade astrocytoma showing abnormal Tc-99m MDP localization. PMID- 9227887 TI - Sesamoids opened earlier and easier by bone imaging. PMID- 9227888 TI - Soft-tissue involvement by adenocarcinoma imaged during bone scintigraphy. PMID- 9227889 TI - Rheological properties of selected dairy products. AB - This article reviews rheological properties of milk, concentrated milk, cream, butter, ice cream, and yogurt, as well as the structure and some physicochemical properties of milk components. A brief description of basic rheological concepts related to liquids, solids, and viscoelasticity is presented, including those rheological models commonly used to characterize dairy products. Rheological behaviors exhibited by these dairy products, including Newtonian in milk and concentrated milk, nonNewtonian in concentrated milk, cream, and yogurt, thixotropy revealed by concentrated milk, cream, and yogurt, and the viscoelastic characteristics displayed by butter, ice cream, and yogurt, are analyzed, and relevant process variables affecting the rheological behavior of dairy products are discussed. Also, to facilitate the comparison of test methods and identify the typical instrumentation and models utilized in rheological characterization of dairy products, experimental conditions and equations used for modeling are included in a tabulated form. PMID- 9227891 TI - Biologically active factors in bovine milk and dairy byproducts: influence on cell culture. AB - Substantial progress has been made in our knowledge of the biological properties of mammal milks. Many nutritional, biochemical, immunological, or other biological properties have been studied in mature or industrially processed bovine milk as well as in human milk and colostrum. This article is a critical review of selected publications covering (1) the use of bovine milk or dairy byproducts (processed acid and enzymatic whey fractions) as a serum substitute for cell cultures, (2) specific factors in bovine milk and industrially processed milk the affect cell proliferation, and (3) the known functional and biological roles of two whey proteins: beta-lactoglobulin and the PP3 component. PMID- 9227890 TI - Composition and characterization of soyabean and related products. AB - Soyabean contains about 48 to 50% proteins. Among these, storage proteins are predominant. 7S and 11S globulins are two storage proteins that constitute 80% of the total protein content in soyabean. Moreover, there are other less abundant storage proteins such as 2S, 9S, and 15S globulins. In addition to globulins, enzymes, protease inhibitors (Kunitz and Bowman-Birk), lectin, and other complete the soya protein content. Different methods exist to characterize soya proteins. These methods involve (1) an isolation of proteins from soya commercial products and (2) the use of analytical techniques for protein determination. Soya proteins may interact with other soya components such as minerals, phytic acid, ascorbic acid, and fiber. These interactions, which depend on soya processing and treatment, can decrease the bioavailability of minerals and proteins. Swelling, solubility, viscosity, and capacity to form a gel, an emulsion, or a foam are the main functional properties of soyabean. They are responsible for the wide use of soya in industrial processes. PMID- 9227892 TI - A 900 bp genomic region from the mouse dystrophin promoter directs lacZ reporter expression only to the right heart of transgenic mice. AB - In order to study the regulatory mechanism of developmental and tissue-specific expression of the muscle type dystrophin gene in mice, transgenic mice were generated carrying the 900 bp genomic fragment derived from the muscle type dystrophin promoter region fused to the bacterial lacZ gene. Six independent transgenic mouse lines showed specific reporter gene expression in the right heart, but not in skeletal or smooth muscle. The reporter gene expression was first detected in the presumptive right ventricle of the embryos at 8.5 days post coitum and the expression continued only in the right ventricle throughout the development and at the adult stage. The results indicate that the 900 bp genomic fragment contains the regulatory element required for expression of dystrophin only in the right heart, suggesting that distinct elements are responsible for the expression in the left and right compartments of the heart, and/or in skeletal and smooth muscle cells. Based on these findings, the relationship between defects in muscle type promoter and the diseases caused by abnormal dystrophin expression is discussed. PMID- 9227893 TI - Differentiation of female chicken primordial germ cells into spermatozoa in male gonads. AB - In avian species, the developmental fate of different-sex germ cells in the gonads is unclear. The present study attempted to confirm whether genetically female germ cells can differentiate into spermatozoa in male gonads using male germline chimeric chickens produced by the transfer of primordial germ cells (PGC), and employing molecular biological methods. As a result of Southern hybridization, specific sequences of the W chromosome (the female specific sex chromosome in birds) were detected in the genomic DNA extracted from one out of four male germline chimeric chickens. When two-color in situ hybridization was conducted on the spermatozoa of this germline chimera, 0.33% (average) of the nuclei of each semen sample showed the fluorescent signal indicating the presence of the W chromosome. The present study shows that female PGC can differentiate into spermatozoa in male gonads in the chicken. However, the ratio of produced W chromosome-bearing (W-bearing) spermatozoa fell substantially below expectations. It is therefore concluded that most of the W-bearing PGC could not differentiate into spermatozoa because of restricted spermatogenesis. PMID- 9227894 TI - An immunocytochemical analysis of the expression of thyroid hormone receptor alpha and beta proteins during natural and thyroid hormone-induced metamorphosis in Xenopus. AB - Amphibian metamorphosis is characterized by the upregulation of thyroid hormone receptor (TR) mRNA in all tissues of tadpole during both the natural and thyroid hormone (TH)-induced development. The two TR genes, termed alpha and beta, are members of a large multigene family of nuclear receptors related to the cellular homolog of the oncogene c-erbA. The phenomenon of upregulation is more marked for the beta than the alpha isoform. To determine whether or not the auto-induction of the transcripts is paralleled by that of TR proteins, non-cross-reacting monoclonal antibodies were prepared against Xenopus laevis TR alpha and beta (xTR alpha, beta) in order to analyze immunocytochemically their expression and localization. Three tadpole tissues that exemplify three major consequences of gene re-programing during natural and TH-induced metamorphosis were studied: (i) Liver that undergoes extensive functional switching; (ii) small intestinal epithelium that exhibits substantial cell death prior to major structural and biochemical modifications; and (iii) hind limb-bud as an example of de novo morphogenesis. It was shown that xTR alpha protein is generally more abundant in these tissues, and its expression is developmentally and hormonally less regulated, than is xTR beta. The auto-induction of xTR beta was particularly intense at 5 days after administration of triiodo-thyronine (T3) to both pre metamorphic (stage 52) tadpoles and at the onset of natural metamorphosis (stage 55). In the developing hind limb-bud at both stages the upregulation of TR beta is topologically restricted, being particularly intense in dense pockets of cells, presumably rich in chondrocytes. It was concluded that the distribution and expression of xTR alpha and beta proteins match partially, but not fully, those of their transcripts during natural and hormone-induced metamorphosis. PMID- 9227895 TI - Spfkh1 encodes a transcription factor implicated in gut formation during sea urchin development. AB - A member of the forkhead class of transcription factors from sea urchins (Spfkh1) that is expressed specifically in the endoderm of developing embryos has been identified. Spfkh1 was expressed transiently in the embryo, with peak levels of messenger ribonucleic acid (mRNA) accumulating at the time endoderm invaginated into the interior of the embryo. Expression was limited to the invaginating endoderm in the early gastrula, then became further restricted to the base of the invaginating gut at the mid-gastrula stage. Expression diminished by the end of gastrulation. This expression pattern indicates that Spfkh1 mRNA accumulates in endodermal cells as they invaginate, but disappears rapidly in endodermal cells that undergo convergent extension. Treatment of embryos during cleavage stages with lithium or phorbol esters caused an increase in Spfkh1 mRNA accumulation and expanded the domain of expression of Spfkh1, suggesting that signaling through the inositol-tris-phosphate protein kinase C (IP3-PKC) signaling pathway is upstream of Spfkh1 expression. The expression pattern of Spfkh1 suggests that it is centrally involved in specification and/or differentiation of the gut. Disruption of the extracellular matrix (ECM) prevents formation of the gut, but does not inhibit initiation of Spfkh1 expression. Embryos arrested prior to gastrulation continued to express Spfkh1 well past the time it was down-regulated in normal embryos, suggesting the ECM or cell movement is required for the decrease in Spfkh1 mRNA during gastrulation. PMID- 9227896 TI - Activation at the germinal vesicle stage of starfish oocytes produces parthenogenetic development through the failure of polar body extrusion. AB - Starfish oocytes can be fertilized after germinal vesicle breakdown (GVBD) and artificial parthenogenesis can be induced by activating the oocytes after GVBD (post-GVBD activation). In the present study, parthenogenotes were obtained by the activation of immature oocytes with caffeine before treatment with 1 methyladenine (1-MeAde) to induce oocyte maturation. Most of the caffeine-treated eggs developed as tetraploids, as parthenogenotes produced by the post-GVBD activation. The parthengenotes were derived only from eggs that failed to extrude polar bodies, mostly from eggs failing to extrude a second polar body. Eggs derived from immature oocytes activated by A23187, treated with 1-MeAde and post treated with cytochalasin B failed to extrude polar bodies, and eventually developed into parthenogenetic embryos. These results indicate that the present parthenogenesis mechanism shares the same characteristics as that achieved by post-GVBD activation in the suppression of polar body formation as a key means for successful starfish parthenogenesis. PMID- 9227897 TI - Quantitative and spatial information on the composition of chimaeric fetal mouse eyes from single histological sections. AB - The spatial distribution of cells in chimaeric tissues, composed of two genotypes, provides insights into the extent of cell mixing during development and growth. However, direct measurement of patch sizes is not usually meaningful because, when the proportion of one genotype is high, a single patch may encompass several adjacent coherent clones of like genotype (clone aggregation). Two previously used methods of comparing patch lengths were evaluated to overcome this problem. The corrected mean patch length (corrected for the predicted effects of random clone aggregation) is a more useful summary statistic than the median patch length of the minor genotype, because its use is not restricted to grossly unbalanced chimaeras, but its validity has been questioned. The two methods gave almost identical numerical summaries of patch sizes in the retinal pigment epithelium of fetal chimaeras, thereby validating the use of the corrected mean patch length for this tissue. The present study also showed that the corrected patch length was unaffected by the presence of cells hemizygous for the TgN(Hbb-b1)83Clo transgene and that the proportion of pigmented cells in a single histological section was representative of the overall composition of the chimaeric fetus. PMID- 9227898 TI - Oral/aboral ectoderm differentiation of the sea urchin embryo depends on a planar or secretory signal from the vegetal hemisphere. AB - A monoclonal antibody that recognizes oral ectoderm and esophagus of sea urchin larvae was newly produced. Distribution of the antigen, named Hpoe, was examined by indirect immunofluorescence microscopy. Hpoe did not exist in eggs and appeared during the cleavage stage. In hatched blastulae, Hpoe was detected on the apical surface of all cells. As embryogenesis progressed, Hpoe disappeared from the primary mesenchyme, archenteron and aboral ectoderm. Hpoe reappeared in foregut at the prism stage and was restricted to the oral ectoderm and esophagus at the pluteus stage. Using this antigen as a molecular marker of oral/aboral ectoderm differentiation, the role of the vegetal hemisphere in ectoderm differentiation was examined. All animal hemispheres isolated from 16-cell stage embryos, mesenchyme blastulae, early gastrulae and mid gastrulae developed into epithelial balls and every cell expressed Hpoe. These epithelial balls failed in oral/aboral ectoderm differentiation. Twenty millimolar LiCl-treated whole embryos developed into exo-gastrulae but Hpoe restriction in ectoderm occurred in these exo-gastrulae. These results show that oral/aboral ectoderm differentiation requires an inductive interaction from the vegetal hemisphere and indicate that the inductive interaction depends on a planar or secretory signal, rather than the contact of the esophagus and ectoderm. PMID- 9227899 TI - A soluble extract from human spermatozoa activates ascidian oocytes. AB - A soluble extract from human spermatozoa induced calcium oscillations and extrusion of the first polar body when injected into oocytes of the ascidian Ciona intestinalis. The properties of calcium oscillations and time of polar body extrusion precisely mimic oocyte activation induced by C. intestinalis sperm or sperm extracts. The data suggest that human sperm extracts can activate oocytes of different phyla by the same mechanism as homologous spermatozoa. Injection of inositol 1,4,5-trisphosphate (IP3) into C. intestinalis oocytes mimicked to some extent the initial stages of oocyte activation, but the results demonstrate that ascidian oocyte activation by human sperm extract cannot be explained solely in terms of IP3-induced calcium release. Injection of other calcium releasing second messengers, cyclic adenosine diphosphate ribose, or calcium ions, does not lead to oocyte activation or release intracellular calcium in ascidian oocyte. It was concluded that human spermatozoa contain one or more molecules than can trigger intracellular calcium release in oocytes from different phyla. PMID- 9227900 TI - Establishment of in vitro spermatogenesis from spermatocytes in the medaka, Oryzias latipes. AB - Spermatocytes of the teleost, Oryzias latipes, at meiotic prophase were cultured without contact with somatic cells. They began to divide, progressing through the meiotic divisions and differentiating into round spermatids within 48 h. The chromosome number in both the primary and secondary spermatocytes at metaphase was n = 24. In spermatids, a single flagellum was formed and the release of residual bodies was observed in vitro. The size and shape of the flagellum were the same as those seen in vivo. The expression of protamine mRNA was detected in round spermatids. This result suggests that gene expression, as well as morphological change, is regulated by the progression of spermatogenesis in cell culture. Furthermore, when the eggs of O. latipes were inseminated with germ cells cultured for 10 days, normal embryos developed and hatched out. These results suggest that the spermatocytes of O. latipes develop into fertile sperm in cell culture. PMID- 9227901 TI - The cause of the decreased number of primordial germ cells in albino Xenopus resides not in the micro-environment but in the presumptive PGC. AB - The number of primordial germ cells (PGC) in albino tadpoles of Xenopus is significantly decreased as compared with that of the wild-type. Whether the decreased number of PGC is caused by the presumptive PGC (pPGC) themselves or the micro-environment surrounding those cells in the albino, or both was investigated in the present study. [3H]thymidine-labeled pPGC of wild-type and albino were implanted into unlabeled, host neurulae of wild-type or albino and wild-type, respectively. Labeled PGC in the genital ridges of experimental tadpoles were examined by autoradiography. There were no significant differences in the proportion of tadpoles with labeled PGC and in the average number of those PGC between the albino and wild-type tadpoles, into which wild-type pPGC had been implanted. The proportion in wild-type tadpoles with albino pPGC was much lower than that in wild-type tadpoles with wild-type pPGC. These results suggest that the pPGC of the albino and not the micro-environment are responsible for the decreased number of PGC. PMID- 9227902 TI - Role of intercellular contacts in generating an asymmetric mitotic apparatus in the Tubifex embryo. AB - The 2-cell stage embryo of Tubifex is composed of a smaller cell, AB, and a larger cell, CD. At the second cleavage, the CD-cell divides unequally. The mitotic apparatus (MA) involved in this division is organized asymmetrically: the MA pole to be segregated to a smaller cell is flattened and truncated, and associated with the anterior cortex facing the AB-cell, while the other pole is symmetric and located more centrally. The present study was undertaken to elucidate the mechanism that generates asymmetry in the MA organization in CD cells. When CD-cell nuclei, which are normally located near the anterior cortex, were displaced toward the posterior end of the cell (i.e. opposite AB-cells) by centrifugation, MA assembled ectopically there, and were bilaterally symmetric in organization. Similar symmetric MA were formed in isolated CD-cells, which divided more equally than intact cells. This equality of cell division was dramatically reduced if the anterior surface of isolated CD-cells formed contact with other cells, such as AB-, C- and 4D-cells. The MA that formed in these reconstituted embryos were asymmetric in organization; one MA pole was always found to be truncated and apposed to the cortical site at the cell contact. Symmetric MA were also observed in cytochalasin-treated embryos. Together with the finding that one of the MA poles is physically attached to the anterior cortex of the intact CD-cell, these results suggest that factors generating asymmetry in the spatial organization of MA poles reside at the anterior cortex of the CD-cell and that this cortical mechanism is dependent upon cell contacts. PMID- 9227903 TI - Isolation of a differentiation-defective myoblastic cell line, INC-2, expressing muscle LIM protein under differentiation-inducing conditions. AB - A non-differentiating myoblastic cell line, INC2, and a differentiating cell line, COM3, were established from the mouse myoblastic cell line C2C12. Under differentiation conditions, both COM3 and INC2 cells stopped proliferation in a similar manner. The COM3 cells then differentiated into myotubes during the 4-day differentiation culture. In contrast, almost none of the INC2 cells differentiated into myotubes even in differentiation medium. Reverse transcriptase-polymerase chain reaction (RT-PCR) and immunoblot analyses showed that the levels of myogenin and MyoD proteins were significantly decreased in INC2 cells. The differentiation marker sarcomeric myosin heavy chain (MHC) was expressed in COM3 but not in INC2 cells. In contrast, both INC2 and COM3 cells expressed another myogenic regulatory factor, muscle LIM protein (MLP), in a differentiation condition-dependent manner. These results suggest that MLP gene expression is regulated in a myogenin/MyoD-independent manner. Enforced expression of the myogenin gene induced MHC expression in INC2 cells. Thus, the signaling pathway situated downstream is assumed to be intact in INC2 cells and suppression of myogenin, gene expression may be a primary defect in INC2 cells. PMID- 9227904 TI - Oral-aboral ectoderm differentiation of sea urchin embryos is disrupted in response to calcium ionophore. AB - Intracellular signaling mediated by calcium ions has been implicated as important in controlling cell activity. The ability of calcium ionophore (A23187), which causes an increase in calcium ion concentration in the cytoplasm, to alter the pattern of differentiation of cells during sea urchin development was examined. The addition of A23187 to embryos for 3 h during early cleavage causes dramatic changes in their development during gastrulation. Using tissue-specific cDNA probes and antibodies, it was shown that A23187 causes the disruption of oral aboral ectoderm differentiation of sea urchin embryos. The critical period for A23187 to disturb the oral-aboral ectoderm differentiation is during the cleavage stage, and treatment of embryos with A23187 after that time has little effect. The A23187 does not affect the formation of the three germ layers. These results indicate that intracellular signals mediated by calcium ions may play a key role in establishment of the oral-aboral axis during sea urchin development. PMID- 9227905 TI - Isolation and characterization of an endodermally derived, proteoglycan-like extracellular matrix molecule that may be involved in larval starfish digestive tract morphogenesis. AB - A monoclonal antibody, anti-Pisaster matrix-1 (anti-PM1) has been developed against an extracellular matrix antigen, Pisaster matrix-1 (PM1) found in embryos and larvae of the starfish Pisaster ochraceus. Pisaster matrix-1 was first observed in endodermal cells of the early gastrula, and shortly thereafter it was secreted into the blastocoel where it accumulated steadily during gastrulation. During the late gastrula stage it also appeared in the extracellular matrix (ECM) of the gut lumen. Immunogold electron microscopy with anti-PM1 revealed that PM1 was found in condensations of ECM associated with blastocoel matrix fibers, in the trans Golgi network, in Golgi-associated vesicles in endoderm and mesenchyme cells and throughout the ECM lining the digestive tract of late gastrula and bipinnaria larvae. When blastula or early gastrula stage embryos were grown in the presence of the PM1 antibody, archenteron elongation, bending and mouth formation failed to occur. Pisaster matrix-1 stained with alcian blue and its assembly could be disrupted with the common inhibitor of O-linked glycosaminoglycan assembly, beta-xyloside but not by tunicamycin. It was not sensitive to enzymes that degrade vertebrate proteoglycans. Pisaster matrix-1 is a large (600 kDa) proteoglycan-like glycosaminoglycan, secreted exclusively by endodermal and/or endodermally derived cells that may be necessary for morphogenesis of the mouth and digestive tract of Pisaster ochraceus embryos/larvae. PMID- 9227906 TI - Mdes, a mouse homolog of the Drosophila degenerative spermatocyte gene is expressed during mouse spermatogenesis. AB - A new mouse gene Mdes has been identified, which has a significant sequence homology with a Drosophila gene degenerative spermatocyte (des) that is required for the initiation of meiosis in spermatogenesis. The expression pattern of the Mdes transcript during mouse spermatogenesis is similar to that of the des transcript curing Drosophila spermatogenesis. Based on these results, it is proposed that the products of Mdes and des have a phylogenetically conserved role in vertebrate and invertebrate spermatogenesis. PMID- 9227907 TI - The response of bovine granulosa cells to different gonadotrophins in culture. AB - Previous studies with bovine granulosa cells cultured in vitro indicated that follicle-stimulating hormone (FSH) stimulated differentiation and progesterone production of granulosa cells in a dose-dependent manner, this was due mainly to an increase in the number of differentiated cells. The objectives of the present study were to investigate (1) whether the response of bovine granulosa cells in culture to luteinising hormone (LH) and equine chorionic gonadotrophin (eCG) was similar to the response to FSH, and (2) whether granulosa cells derived from different cattle breeds responded similarly to gonadotrophin stimulation. Pairs of ovaries were recovered postmortem from Charolais (38) and Hereford (41) crossbred post-pubertal heifers, and granulosa cells were aspirated from 5-8 mm follicles. In two simultaneous experiments, granulosa cells (2-3 x 10(5) viable cells) were cultured with different gonadotrophins (oFSH or oLH in Experiment 1; oFSH or eCG in Experiment 2). Cell culture was for 4 days at 37 degrees C in a humidified atmosphere of 5% CO2 in air in 1 ml of serum-free culture medium. Progesterone production, total DNA and the protein content of granulosa cells on Day 4 of culture were determined. Log10 data were analyzed by analysis of variance and multiple linear regression. In Experiment 1, both FSH and LH stimulated progesterone production (ng microgram-1 DNA) and protein content (microgram microgram-1 DNA) of granulosa cells in a dose-dependent manner (P < 0.01). The relative potencies of FSH to LH (milli micron/milli micron) were found not to be different from unity. In Experiment 2, progesterone production and the protein content of granulosa cells were stimulated by both FSH and eCG in a dose dependent manner (P < 0.001). The progesterone response curves (log/log) were linear up to 1-10 milli microns FSH and 1-10 iu eCG, and were Y = 1.67 + 0.093 FSH and Y = 1.60 + 0.091 eCG for progesterone production. Calculated on a milli micron/iu basis, FSH was found to be 5.8 times more potent than eCG (P < 0.05) in terms of stimulating progesterone production. Granulosa cells derived from Hereford crosses were more sensitive (P < 0.001) than those from Charolais crosses to gonadotrophin stimulation (31 and 42 times for FSH and eCG, respectively, in terms of progesterone production, and 4.8 and 3.1 times for FSH and eCG, respectively, in terms of protein content). The response curves for both FSH and eCG were similar within each breed. The slopes of the progesterone response curves, and the protein responses were similar for all the gonadotrophins. In conclusion, these results imply that FSH; LH and eCG have similar effects on the differentiation and progesterone production of bovine granulosa cells from 5-8 mm follicles cultured in vitro. Furthermore, granulosa cells from different breeds cultured in vitro had different sensitivities to gonadotrophin stimulation. PMID- 9227908 TI - Oestradiol potentiates a prolonged progesterone-induced suppression of LH release in ovariectomised cows. AB - Our working hypothesis was that the negative feedback effect of progesterone on LH secretion in ovariectomised cows is greatly enhanced by oestradiol when both hormones are administered intravaginally. Each of eight ovariectomised cows had a progesterone releasing device ('CIDR-1'; Eazi-breed CIDR-B. InterAg, Hamilton, New Zealand) inserted into the vagina for 3 days, followed by an injection of 1 mg oestradiol benzoate (ODB) in peanut oil 24 h after device removal. The timing of this injection was designated as Day 0. The CIDR-1 was reinserted concurrently with an injection of 0.4 mg ODB on Day 5. An additional device ('CIDR-2') was inserted 4 days later (Day 9). In four of the cows, an ODB capsule (CIDIROL), Douglas Pharmaceuticals. Auckland, New Zealand) was placed into a groove on the surface of CIDR-2 preceding insertion. CIDR-2 was withdrawn after 5 days (Day 14) and CIDR-1 was withdrawn 2 days later (Day 16). The treatment regimen was repeated in a cross-over design where each cow received CIDR-2 with or without an ODB capsule. Blood samples were collected at intervals of 15 min for a period of 16 h beginning 8 h before insertion of CIDR-2 to determine changes in LH secretion pattern, and then at intervals of 8 h until 48 h after removal of CIDR 1 to determine mean LH and steroid concentrations. Concentrations of progesterone in plasma reflected the effects of inserting and removing each CIDR device. The insertion of CIDR-2 elevated progesterone from 2.0 +/- 0.1 ng ml-1 to 4.9 +/- 0.1 ng ml-1 within 30 min (P < 0.01) and maintained levels > 3 ng ml-1. Withdrawal of CIDR-2 produced a new baseline level of 1.8 +/- 0.1 ng ml-1 within 4 h (P < 0.01). Concentrations of oestradiol-17 beta in plasma increased from a baseline of < 1 pg ml-1 to-a-maximum of 12.6 +/- 2.2 pg ml-1 at 22 h after insertion of an ODB capsule. Initial pulse frequency of LH (1.2 +/- 0.05 pulses h-1) and mean concentration of LH (1.6 +/- 0.1 ng ml-1) declined (P < 0.05) to 0.2 +/- 0.1 pulses h-1 and 0.5 +/- 0.1 ng ml-1 during the period from 2 to 8 h following insertion of CIDR-2. Mean concentrations recovered to pre-treatment levels (> 1 ng ml-1) within 48 h if CIDR-2 was inserted without an ODB capsule. In contrast, LH levels remained less than 0.7 ng ml-1 for 5 days following this form of ODB treatment. A subsequent increase in mean LH of 1.4 +/- 0.2 ng ml-1 (P < 0.05) was inversely synchronised with the precipitous decline in progesterone of 1.2 +/- 0.1 ng ml-1, following removal of CIDR-2. The inclusion of an ODB capsule with insertion of CIDR-2 reduced concentrations of FSH from 166 +/- 17 ng ml-1 to a new baseline of 105 +/- 16 ng ml-1 within 24 h (P < 0.05). In conclusion, an acute increase in progesterone induced a transient suppression of LH lasting less than 48 h. This suppression was prolonged to 5 days by the concurrent insertion of a capsule containing 10 mg ODB. The subsequent recovery of LH was coincident with the decline in progesterone following removal of CIDR-2. PMID- 9227909 TI - Development of in vitro derived bovine embryos following pronuclear transplantation and in vitro culture. AB - This study was designed to evaluate the survival and development of in vitro derived bovine embryos following pronuclear transplantation and in vitro embryo culture. Bovine zygotes were produced by in vitro maturation and in vitro fertilization. Pronuclei were removed by micromanipulation and either transferred back to the same cell (Group 1) or into a previously enucleated zygote (Group 2) by electrofusion. Micromanipulated and non-micromanipulated (Group 3, control) zygotes were co-cultured with oviductal cells in a sealed modular chamber filled with 5% CO2, 5% O2 and 90% N2 at 39 degrees C for 7-8 days. Fusion rates were similar for Groups 1 and 2 (90.7 and 85.1%, respectively, P > 0.05). The percentage of embryos that cleaved was not different for Groups 1 (82.0%), 2 (90.0%) and 3 (76.9%, P > 0.05). Also, the percentage of embryos developing to the compact morula or blastocyst stage was similar (25.6, 22.5 and 22.3%, respectively, for Groups 1, 2 and 3, P > 0.05). The results of this experiment are the first to demonstrate that pronuclear transfer can be carried out successfully using bovine embryos derived from in vitro oocyte maturation and in vitro fertilization. In addition, pronuclei can be transferred from one bovine embryo to another and the reconstructed embryos develop to the compact morula and blastocyst stage in vitro. This technique, used in combination with oocyte retrieval by ultrasound-guided follicular aspiration and embryo transfer, offers the potential to study cytoplasmic inheritance in cattle directly, and to evaluate the effect of cytoplasmic inheritance on traits of economic importance. PMID- 9227910 TI - In vitro maturation of bovine oocytes in the presence of bovine activin A does not affect the number of embryos. AB - This study was carried out to investigate whether bovine recombinant activin A present during the in vitro maturation (IVM) of cumulus enclosed bovine oocytes affects the proportion of embryos that develop to the blastocyst stage. In addition, the effect of the presence of activin A during maturation and during embryo culture was studied. Therefore, bovine cumulus oocyte complexes were matured at 39 degrees C in a humidified atmosphere with 5% CO2 in air for 24 h in: (1) culture medium M199 supplemented with 10% foetal calf serum (FCS), luteinising hormone (LH) and follicle-stimulating hormone (FSH) and 10 ng ml-1 activin A; (2) M199 without FCS but supplemented with LH and FSH and 10 ng ml-1 activin A; (3) M199 without FCS, LH and FSH but supplemented with 10 ng ml-1 activin A. Cultures without activin served as controls. After IVF the embryos were cultured in M199 supplemented with 10% FCS on a monolayer of buffalo rat liver (BRL) cells. For the second part of the study, COCs were matured in vitro in M199 supplemented with LH and FSH and 10 ng ml-1 activin A, fertilized in vitro and the embryos were cultured (1) on a monolayer of BRL cells in M199 supplemented with 10% FCS and 10 ng ml-1 of activin A, and (2) in droplets of serum free BRL-conditioned medium supplemented with 10 ng ml-1 activin A. IVM in the presence of LH, FSH and 10 ng ml-1 activin A did not change the proportion of blastocysts present at Day 9 or the proportion of hatched blastocyst at Day 11. Activin present during maturation in the absence of serum and gonadotrophic hormones also did not alter the proportion of blastocysts or hatched blastocysts. In vitro culture of embryos on BRL cells or in BRL-conditioned medium in the presence of activin had no effect on embryonic development. It is concluded that IVM in the presence of bovine activin A has no effect on subsequent embryonic development. PMID- 9227911 TI - Oestrous behaviour of Holstein cows during cooler and hotter tropical seasons. AB - Seasonal effects on post-partum ovarian activity, duration and intensity of sexual behaviour were determined for Holstein dairy cattle imported from a temperate climate into a tropical region. Animals were observed continuously during the cooler (temperature-humidity index (THI) < 25) and hotter (THI > 25) seasons for 2 years. They were restricted to a cement footing in the hotter season observation period in Year 1, but had access to both concrete and dirt footing during all other seasons. Sequential milk progesterone profiles provided an indication of when follicular phases occurred, and recorded sexual behaviour was compared with these to determine if oestrous signs accompanied ovulations. Most cows had normal ovarian cycles and ovulated regularly during both seasons, but quiet ovulations occurred with greater frequency during the hotter times of the year (P < 0.05). Demonstrations of sexual behaviour were affected by choice of footing rather than season. The actual time when cows stood passively and allowed herdmates to complete mounting ranged from 5.1 +/- 0.7 to 5.8 +/- 1 h with access to exercise yards and cement or dirt footing, but declined to only 1.3 +/- 1.1 h when animals were confined to cement (P < 0.05). Similarly, the total duration of oestrus and mean number of interactions were significantly (P < 0.05) reduced during the observation period conducted with cows confined to concrete footing. These findings further emphasize that the duration of oestrus in dairy cows in considerably shorter than the commonly quoted 18 h. PMID- 9227913 TI - Evidence that the conceptus contributes to the inhibition of follicular growth in the ewe. AB - The present study was designed to determine the contribution of the gravid uterus and the conceptus in the inhibition of antral follicular growth. All antral follicles were counted and measured on day 48 post mating in ovaries of pregnant ewes (Group 1, n = 5) and in ovaries of ewes from which the uterus (Group 2, n = 5) or the conceptus (Group 3, n = 5) had been surgically removed on days 32-33 post mating. The secretory activity of the corpora lutea was maintained after removal of the uterus or the conceptus until slaughter of ewes. No difference was found between the three groups of ewes in the mean (+/-SEM) total number (non atretic and atretic) of follicles per ewe, being 59.8 +/- 7.6, 35.0 +/- 7.6 and 50.8 +/- 4.6, respectively for Groups 1, 2 and 3. The mean (+/-SEM) number of follicles 2 mm or more in diameter was greater (P < 0.01) in Group 2 ewes (3.4 +/ 0.5) and Group 3 ewes (4.2 +/- 0.5) than in Group 1 ewes (1.6 +/- 0.4). Of all follicles counted which were 2 mm or larger, 90% in Group 1 were atretic, this value being higher (P < 0.05) than in Group 2 (68%) and Group 3 (55%). The mean (+/-SEM) diameter of the largest (F1) and second largest (F2) follicles was greater (P < 0.01) in Group 2 (F1, 4.0 +/- 0.4; F2, 3.2 +/- 0.4) and in Group 3 (F1, 3.9 +/- 0.2; F2, 3.3 +/- 0.2) than in Group 1 (F1, 2.6 +/- 0.1; F2, 2.2 +/- 0.1). The mean diameter (+/-SEM) of the largest non-atretic follicles was greater (P < 0.01) in Group 2 (4.0 +/- 0.4) and Group 3 (3.9 +/- 0.2) than in Group 1 (1.7 +/- 0.3). In conclusion, the results from this in vivo experiment provide evidence that the conceptus prevents the development of large healthy follicles by inducing atresia during the first trimester of pregnancy. The inhibition of follicular growth would reduce oestradiol production and permit the maintenance of luteal function during early pregnancy. PMID- 9227912 TI - Effect of nutrition on the balance of production of ovarian and pituitary hormones in ewes. AB - Nutrition-induced changes in liveweight can induce changes in ovarian function and ovulation rate. This study was designed to test the hypothesis that heavy ewes are able to maintain similar concentrations of follicle-stimulating hormone (FSH) to light ewes through a loss of responsiveness to ovarian negative feedback. This hypothesis leads to the prediction that administration of ovarian hormones on the basis of metabolic body weight would result in the same gonadotrophin (FSH) concentrations in-light and heavy ovariectomised ewes. We therefore examined the ovarian content of oestradiol and inhibin, and the FSH response to administration of these ovarian hormones in acutely ovariectomised heavy and light Merino ewes produced by differential feeding. At ovariectomy, follicular fluid was aspirated from the 6 largest follicles in each ewe and assayed for oestradiol and inhibin. Six days later, blood was sampled every 6 h for 2 days. Ewes were then given two subcutaneous progesterone implants and injected subcutaneously every 6 h for 5 days with oestradiol (37.4 ng kg-0.75) and charcoal-stripped bovine follicular fluid (0.04 ml kg-0.75). Blood was sampled before each injection and the plasma was assayed for FSH and inhibin. Oestradiol and its receptors were assayed in uterine tissue. Nutrition-induced increases in liveweight led to increases in the number of oestrogenic, potentially ovulatory ovarian follicles and the total ovarian content of oestradiol and inhibin. In the uterus, more oestradiol receptors and a higher oestradiol uptake were observed in light than in heavy ewes. After ovariectomy, FSH concentrations were inversely related to liveweight. However, when the ewes were treated with doses of ovarian hormones that were proportional to liveweight, the slope of the decline in FSH concentrations was independent of liveweight. We conclude that nutrition alters the balance between pituitary FSH secretion and gonadal feedback by changing the responsiveness to the inhibitory effects of oestradiol and inhibin. PMID- 9227914 TI - Changes in sperm-bound amidase activity suggest subtle damage to ram sperm acrosomes by freezing/thawing, not detected by light microscopy. AB - We have measured sperm-bound amidase activity in fresh, cooled and frozen/thawed ram spermatozoa, in order to study if freezing and thawing led to some degree of acrosome damage of motile/viable spermatozoa not detected by optical methods. This assay was based on the fact that membrane damage would result in an increased access of the enzyme substrate to the sperm acrosome. Semen was collected from adult Australian Merino rams, and spermatozoa were washed by centrifugation through a Ficoll solution. Sperm-bound amidase activity was measured in whole spermatozoa using the protease substrate benzoyl-arginyl-p nitroanilide (BAPNA). Acrosomal status was also assessed by light microscopy after Giemsa staining. Most amidase activity was shown to be sperm-bound, as only a minor fraction of the enzyme activity was release into the medium after induced damage. Simultaneous assessment of sperm-bound amidase activity and the percentage of spermatozoa with microscopically evident acrosomal damage, after mild sonication for different times, showed a high correlation between both parameters (r = 0.97, p < 0.001). In separate experiments, fresh, cooled and frozen/thawed semen samples were filtered through Sephadex G-10 to obtain a subpopulation of motile, mostly acrosome-intact spermatozoa. As controls, spermatozoa from the same samples to which extensive acrosome damage was induced were evaluated. Slow cooling to 4 degrees C had no effect on amidase activity or percent acrosomal damage with respect to fresh samples. Freezing and thawing resulted in a sperm population that, after filtration through Sephadex, had a low percentage of acrosome damage (9.4%, vs. 2.1% for fresh filtered controls), which was 11% of that obtained after extensive acrosome damage (83%). However, amidase activity in these samples was markedly increased, showing values of activity that were 56% of those obtained in extensively damaged spermatozoa. This effect was not due to an alteration in the enzyme kinetics. We conclude that sperm-bound amidase activity is useful to detect subtle changes, provoked by a standard freezing/thawing procedure, in the permeability of acrosomes from ram spermatozoa which are not detected by direct observation of the acrosomes after Giemsa staining. PMID- 9227915 TI - Fibronectin concentrations correlate with ovarian follicular size and estradiol values in equine follicular fluid. AB - The amounts of total protein, albumin, fibronectin, alpha 2-macroglobulin (alpha 2-M), immunoglobulin G, ceruloplasmin and antithrombin were determined in fluids collected from 53 preovulatory equine follicles and compared with the contents of estradiol-17 beta, progesterone and androstenedione, with follicle size and the amounts of the equivalent proteins in normal equine plasma. The concentration of fibronectin and the fibronectin/albumin ratios increased significantly with follicle size and with follicular estradiol levels. The alpha 2-M levels and alpha 2-M/albumin ratios correlated with follicle size but not with hormone content. Both fibronectin and alpha 2-M were present in lower amounts in follicular fluid compared with plasma while the other proteins were present in similar amounts. Among the proteins evaluated, there was a positive correlation between the amount of the protein in the follicular fluid and the molecular weight of the protein. PMID- 9227916 TI - Investigations of BrdU incorporation in roe deer blastocysts in vitro. AB - These data provide direct evidence for slow proliferation activity in pre implantation roe deer trophoblasts and even minor growth of the embryoblast. The blastocyst grows very slowly from August to January. Incorporation of BrdU into the nuclei during a period of 4 h was used to determine the proliferation of the embryonic cells in vitro. The results show for the first time the DNA synthesis in trophoblast cells of roe deer blastocysts from October to December. During the period of October to November the incorporation of BrdU is weak, and in December there is a duplication of DNA metabolism. PMID- 9227917 TI - Distribution, number and membrane integrity of spermatozoa in the pig oviduct in relation to spontaneous ovulation. AB - The pattern of distribution, number and membrane integrity of spermatozoa in the utero-tubal junction (UTJ) and isthmus during three oestrous stages were related to spontaneous ovulation in flushed and fixed oviducts of multiparous sows. Three unrelated boars were each used once to mate or artificially inseminate (neat ejaculate) six out of 18 sows, 18 h prior to expected ovulation. The sows were slaughtered 6-8 h before, during or 6-8 h after ovulation. The ad-uterine oviductal region (UTJ and isthmus) was divided into UTJ, lower isthmus, middle isthmus and upper isthmus segments. A higher fraction of middle and upper isthmus segments contained spermatozoa during the peri- and post-ovulatory periods than during the pre-ovulatory period. The distribution, numbers and membrane integrity of spermatozoa in the UTJ-isthmus region were influenced by the ovulation event. Numbers and distribution of spermatozoa varied depending on the boar used. The flushing technique allowed a better assessment of the distribution, number and membrane integrity of tubal spermatozoa than in situ observation with a scanning electron microscope (SEM). PMID- 9227918 TI - The concentration of GnRH in hypothalamus, LH and FSH in pituitary, LH, PRL and sex steroids in peripheral and ovarian venous plasma of hypo- and hyperthyroid, cysts-bearing gilts. AB - The aim of this work was to investigate the hormonal pattern in hypo- and hyperthyroid gilts with experimentally induced cystic ovarian disease (COD). A total of 70 adult, nulliparous gilts divided into six groups were used for the experiment. Group I was euthyroid and control. Group II was made hypothyroid by oral administration of methylthiouracyl for 24 days. Group III represented euthyroid gilts injected with gonadotropins (PMSG and hCG). Group IV consisted of hypothyroid gilts injected with gonadotropins. Group V was treated with L thyroxine for 24 days and Group VI with thyroxine and with gonadotropins for the last 10 days of the test. The treatment of all groups was terminated on the 4th 5th day of the next estrous cycle. The peripheral blood of the gilts was collected on Day 0, and on Day 24. On the 25th day the gilts were laparotomized and cannulas were inserted into utero-ovarian veins of each ovary for blood collection. Simultaneously, peripheral blood samples were collected during 1 to 3 consecutive days. The animals were then slaughtered and the hypothalamus, pituitary and ovaries were frozen and preserved for further analyses. In hypothalamic tissue the content of GnRH: in the pituitary the concentration of LH and FSH; and in peripheral and ovarian blood plasma the level of LH, PRL, E1, P4, A4, T and cortisol (Cl) were estimated by RIA procedure. The level of GnRH in the hypothalamus, and LH and FSH in the pituitary showed a tendency to parallel with thyroid function which may indicate a role of this gland in their production or secretion. In hypothyroid animals an increase of LH and PRL and a slight decrease of secretory function of the ovaries were noted. Injections of gonadotropins in euthyroid or hypo- and hyperthyroid gilts intensified the function of the ovaries, which was manifested by numerous follicular cysts and corpora lutea. The hormonal milieu of gilts from these groups showed a low level of LH, PRL and an increased content of sex steroids in peripheral and ovarian blood. The ovarian steroidogenesis of cyst-bearing gilts was disturbed, which was indicated by an increased level of E1, P4, A4, T, and Cl, but a low level of E2. These disturbances in steroidogenesis in cystic gilts may be caused by a deficiency in LH secretion as the consequence of the pituitary gonadotropin suppression by the used gonadotropins. The steroid hormone pattern of cyst-bearing gilts strongly resembles the endocrine profile noted in polycystic ovarian disease in women. PMID- 9227919 TI - The effect of removing surface-associated proteins from viable chicken spermatozoa on sperm function in vivo and in vitro. AB - When chicken spermatozoa were diluted in isotonic or hypertonic solutions, sperm surface-associated proteins were lost to the medium, quantitatively more protein being lost to medium of higher osmotic strength. The proteins which were removed from the spermatozoa were also found within chicken seminal plasma, but at different relative concentrations, thus demonstrating selective association of only certain seminal proteins with the spermatozoa. The removal of these proteins occurred with only minimal damage to the spermatozoa, as judged by sperm motility, ATP content and ability to exclude eosin. Spermatozoa treated with hypertonic solutions were unable to reach the freshly ovulated egg in the infundibulum and could not be found within the uterovaginal sperm storage tubules after uterovaginal insemination. However, if inseminated directly into the uterovaginal junction, spermatozoa were able to enter the sperm storage tubules, suggesting that the treatment limited their ability to migrate through the vagina. Loss of sperm fertilizing ability following simple centrifugation and washing treatments appears to result from removal of surface-associated proteins. PMID- 9227920 TI - Temporal aspects of ovarian follicular growth and steroidogenesis following exogenous follicle-stimulating hormone in Angus heifers. AB - Ultrasonography and endocrine assay techniques were used to monitor structural and hormonal alterations made by the ovary in response to the biological actions of pituitary-derived follicle-stimulating hormone (FSH-P). Angus heifers (n = 36) were allotted to receive injections (twice per day) of either FSH-P (up to a total of 28 mg over a maximum of 4 days beginning on Day 10 of a synchronized estrous cycle) or saline in order to quantify temporal relationships among follicle growth and steroid hormone profiles. Transrectal ultrasonography was utilized at 12-h intervals to monitor and record follicle growth. Plasma was collected every 12 h for the first 48 h of the experiment and then every 6 h for the remainder of the experiment. At 48 and 60 h after the onset of treatments, prostaglandin F2 alpha (PGF2 alpha; 25 mg) was administered (i.m.). FSH-treated heifers (n = 6 at each time) were terminated at 24, 48, 72 and 96 h following the onset of treatment. Saline-treated heifers were terminated at 24 and 96 h (n = 6 at each time). After ovaries were obtained, follicular number and size were recorded and follicular fluid (FF) was collected. Plasma concentration of progesterone (P) and estradiol (E2) and FF concentration of P, E2, estrone, testosterone and androstenedione were determined by radioimmunoassays. Plasma concentration of E2 increased (P < 0.05) within 36 h of initiation of FSH treatment. Plasma P decreased (P < 0.0001) by 12 h post-PGF2 alpha. Ultrasonographic examination revealed a significant decrease in the number of small follicles by 48 h, whereas the number of medium follicles increased (P < 0.05) by 60 h after the initiation of FSH treatment. The number of large follicles (LF > or = 10 mm diameter) increased (P < 0.01) over the course of the experiment. The total number of ovarian follicles (TF) 24 h after the start of FSH treatment was correlated (r = 0.99; P < 0.0001) with the number of small follicles (SF < or = 5 mm). At 72 h after the onset of FSH treatment, the number of medium follicles (i.e. 6-9 mm) was correlated with TF (r = 0.97; P < 0.0001). Estradiol was the predominant FF steroid. Follicular fluid E2 was greatest in follicles at 72 h after FSH treatment. Follicular fluid E2 and plasma E2 were positively correlated (r = 0.66; P < 0.001). Follicular aromatase activity was estimated by evaluating the ratio of FF estrogens (E) to androgens (A). Elevated aromatase activity (E:A ratio > 1.0) was detected in 196 of 206 follicles. The estrogen to progesterone ratio was used as an estimate of follicle viability. Eighty-five percent of the follicles were estimated to be viable (E:P ratio > 1.0). The peak E:A ratio in LF preceded by 24 h the peak concentration in FF E2 and plasma E2. In MF and SF the E:A ratio increased by 72 h. Enhancement of ovarian follicular growth (i.e. increased number and size of follicles; increased steroidogenesis) by exogenous, pituitary-derived FSH is characterized by (1) increased activity of aromatase, and (2) accumulation of FF E2, events which temporally preceded the increase in plasma concentration of E2. These observations will aid efforts to incorporate recombinant bovine FSH and somatotropin in an effort to develop more predictable superstimulation and ovulation induction protocols. PMID- 9227922 TI - Factors affecting survival rates of in vitro produced bovine embryos after vitrification and direct in-straw rehydration. AB - The aim of this work was to investigate the possibilities of simplification, and to outline the limits of application, of a vitrification method for cow embryos. Morulae and blastocysts were produced by in vitro fertilization of slaughterhouse derived, in vitro matured oocytes with frozen-thawed bull semen, and subsequent culture on a granulosa cell monolayer. Vitrification was performed by equilibration of embryos with 12.5% ethylene glycol and 12.5% dimethylsulphoxide at 20-22 degrees C for 60 s, then with 25% ethylene glycol and 25% dimethylsulphoxide at 4 degrees C for another 60 s. Embryos were then loaded in straws, placed in liquid nitrogen vapour for 2 min, and then plunged. Straws were thawed in a 22 degrees C water-bath, the embryos were directly rehydrated and further incubated in straw, and were then expelled and cultured in vitro for 72 h. In the first experiment, embryos of different age and developmental stage (Day 5 compacted morulae, Day 6 early blastocysts, Days 6 and 7 blastocysts, Day 7 expanded blastocysts and Day 8 hatched blastocysts) as well as Days 7 and 5 blastocysts previously subjected to partial zone dissection were vitrified. After thawing, the re-expansion rates of blastocysts and zona-dissected embryos did not differ (67 and 87%, respectively), and hatching was more frequent for blastocysts frozen in advanced developmental stages (34, 47 and 63% for early blastocysts, blastocysts and expanded blastocysts, respectively). The re-expansion rate of morulae was lower (10%) and no hatching of these embryos was observed. In the second experiment, Day 7 expanded blastocysts were vitrified using PBS, PBS+albumin, TCM199 and TCM199+calf serum as holding media. No differences in re expansion and hatching rates were seen. However, when incubation with the concentrated cryoprotectant solution was performed at 20-22 degrees C, the embryo survival rate decreased (PBS+albumin) or no embryo survived (TCM199+calf serum) the vitrification procedure. In the third experiment, Day 7 expanded blastocysts were vitrified, thawed, cultured for 1 day, and then re-expanded embryos were again vitrified and thawed. Out of the 87% that survived the first cycle, 73% re expanded and 47% hatched following the second vitrification and thawing. These observations prove that the vitrification procedure described is relatively harmless, that it can be used for blastocysts of different developmental stages and that an intact zona is not required to obtain high survival rates. PMID- 9227921 TI - Normal or induced secretory patterns of luteinising hormone and follicle stimulating hormone in anoestrous gonadotrophin-releasing hormone-immunised and cyclic control heifers. AB - The objective was to determine the effect of gonadotrophin-releasing hormone (GnRH), GnRH analogue (GnRH-A) or oestradiol administration on luteinising hormone (LH) and follicle-stimulating hormone (FSH) release in GnRH-immunised anoestrous and control cyclic heifers. Thirty-two heifers (477 +/- 7.1 kg) were immunised against either human serum albumin (HSA; controls; n = 8), or a HSA GnRH conjugate. On day 70 after primary immunisation, control heifers (n = 4 per treatment; day 3 of cycle) received either (a) 2.5 micrograms GnRH or (b) 2.5 micrograms of GnRH-A (Buserelin) and GnRH-immunised heifers (blocked by GnRH antibody titre; n = 6 per treatment) received either (c) saline, (d) 2.5 micrograms GnRH, (e) 25 micrograms GnRH or (f) 2.5 micrograms GnRH-A, intravenously. On day 105, 1 mg oestradiol was injected (intramuscularly) into control (n = 6) and GnRH-immunised anoestrous heifers with either low (13.4 +/- 1.9% binding at 1:640; n = 6) or high GnRH antibody titres (33.4 +/- 4.8% binding; n = 6). Data were analysed by ANOVA. Mean plasma LH and FSH concentrations on day 69 were higher (P < 0.05) in control than in GnRH-immunised heifers (3.1 +/- 0.16 vs. 2.5 +/- 0.12 ng LH ml-1 and 22.5 +/- 0.73 vs. 17.1 +/- 0.64 ng FSH ml-1, respectively). The number of LH pulses was higher (P < 0.05) in control than in GnRH-immunised heifers on day 69 (3.4 +/- 0.45 and 1.0 +/- 0.26 pulses per 6 h, respectively). On day 70, 2.5 micrograms GnRH increased (P < 0.05) LH concentrations in control but not in GnRH-immunised heifers, while both 25 micrograms GnRH and 2.5 micrograms GnRH-A increased (P < 0.05) LH concentrations in GnRH-immunised heifers, and 2.5 micrograms GnRH-A increased LH in controls. FSH was increased (P < 0.05) in GnRH-immunised heifers following 25 micrograms GnRH and 2.5 micrograms GnRH-A. Oestradiol challenge increased (P < 0.05) LH concentrations during the 13-24 h period after challenge with a greater (P < 0.05) increase in control than in GnRH-immunised heifers. FSH concentrations were decreased (P < 0.05) for at least 30 h after oestradiol challenge. In conclusion, GnRH immunisation decreased LH pulsatility and mean LH and FSH concentrations. GnRH antibodies neutralised low doses of GnRH (2.5 micrograms), but not high doses of GnRH (25 micrograms) and GnRH-A (2.5 micrograms). GnRH immunisation decreased the rise in LH concentrations following oestradiol challenge. PMID- 9227923 TI - Comparison of seasonal changes in reproductive parameters of adult male European fallow deer (Dama dama dama) and hybrid Mesopotamian x European fallow deer (D. d. mesopotamica x D. d. dama). AB - In a study, aimed at comparing seasonal reproductive development of European fallow deer (Dama dama dama) with Mesopotamian (D. d. mesopotamico) x European F1 hybrids, five adult males of each genotype, which had been raised together since birth, were maintained as a bachelor group. Morphometric (body weight, neck circumference and testis diameter), endocrine (plasma testosterone concentrations) and seminal (ejaculate volume, spermatozoa per ejaculate and spermatozoa motility) parameters were recorded at fortnightly or monthly intervals for a 15-month period, and antler status was noted daily during the general periods of casting and velvet stripping. In addition, two bucks of each genotype were blood sampled via indwelling jugular catheters every 30 min for 24 h periods on five occasions (2-3 months intervals) during the year, and plasma was analysed for concentrations of testosterone and LH. Parameter profiles of the two genotypes were compared by global and time series ante-dependence covariance analysis to investigate overall profile similarity and the seasonal nature of any observed differences. Plasma hormone profiles from high-frequency blood sampling were subjected to PULSAR analysis to determine pulse frequency and amplitude. Throughout the study hybrid males were approximately 30% heavier than European males. However, both genotypes exhibited dramatic but parallel patterns of body weight change (global P = 0.054). Neck circumference was correlated with body weight throughout (P < 0.05), with similar regression slopes between the genotypes at any sampling time (P > 0.10). Covariance adjustment to a common initial body weight was performed to eliminate the effects of large body weight differences on muscle hypertrophy and regression. While profiles of corrected neck circumference were significantly different at the global level (P < 0.01), analysis by time revealed differences occurring only during the latter period of muscular regression in spring. However, profiles of other parameters, including testis diameter, plasma testosterone concentrations, spermatozoa per ejaculate and percentage motile spermatozoa, exhibited significant displacement between genotypes (global P < 0.05) evident as 2-4 weeks advancement in the sexual development (late summer/autumn) and quiescence (spring) phases for hybrid males relative to European males. Furthermore, mean dates of antler casting and velvet stripping were significantly earlier by 2-3 weeks for hybrid males than European males (P < 0.05). High frequency blood sampling revealed markedly seasonal patterns of secretion of testosterone and LH, with hybrid males exhibiting an apparent earlier onset of high-amplitude testosterone 'surges' in February (late summer) compared to those occurring in April (autumn) for European males. When viewed collectively, the data indicate strongly that the Mesopotamian influence is evident in the earlier attainment of sexual development and fertility in late summer and autumn, and earlier onset of sexual quiescence in spring. This is in accord with anecdotal information on earlier reproductive patterns in purebred Mesopotamian fallow deer. PMID- 9227924 TI - Effect of estrogen-treated porcine ampulla oviductal epithelial cells on early embryonic development in vitro and characterization of their protein synthetic activity. AB - Recent studies by Buhi et al. have demonstrated that estrogen (E2) is responsible for the induction of de novo synthesis and secretion of certain oviductal secretory proteins (OSP) and inhibition of other OSP in porcine oviductal explant cultures. The present work was undertaken to evaluate the effect of E2-treated oviductal epithelial cell coculture on the development of early porcine embryos derived from in vitro matured and fertilized oocytes. In vitro synthesis of secretory proteins by E2-treated oviductal cells used for coculture was also investigated by one-dimensional (1D) and two-dimensional (2D) sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE). The results showed that the cleavage rate was significantly enhanced by coculturing fertilized eggs with E2-treated oviductal epithelial cells. The in vitro protein synthetic pattern of oviductal secretory proteins was influenced by E2 treatment. These variations included the disappearance of one protein (82,000 M(r)) and the appearance of another (33,000 M(r)) in the E2-treated group as assessed by 1D-SDS-PAGE. Additional proteins of M(r) 97,000 and an M(r) 36,000-45,000 complex were increased in abundance by the E2 treatment. Analyses by 2D-SDS-PAGE revealed three major E2-dependent proteins, of M(r) 45,000 (pI 5.5), 43,000 (pI 5.5) and a 36,000-45,000 M(r) (pI 4.8) protein complex, whereas polypeptides of M(r) 97,000 (pI 5.1), 36,000 (pI 8.0) and 25,000 (pI 6.8) were inhibited by E2 treatment. The results demonstrated that porcine epithelial cell protein synthetic patterns are influenced by E2 treatment and that estradiol treatment of oviductal cells may increase the rate of zygote cleavage during early development in vitro in pigs. PMID- 9227925 TI - Development of bovine and porcine embryonic teratomas in athymic mice. AB - Inner cell masses (ICM) and embryonic discs from bovine and porcine blastocysts of various ages were transplanted under the kidney capsule of athymic (nude) mice to evaluate growth of teratocarcinomas containing both differentiated tissues and undifferentiated stem cells. Inner cell masses were isolated immunosurgically from Day 8, Day 9 and Day 10 porcine blastocysts and from Day 8, Day 10 and Day 12 bovine blastocysts. Embryonic discs were mechanically dissected from Day 11 and Day 12 porcine embryos and from Day 14 bovine embryos. Day 6 egg cylinders were dissected from BALB/C embryos and from hybrid embryos of a cross between BALB/C and an outbred strain of mouse. Two to four ICM, embryonic discs or egg cylinders were transplanted under the kidney capsule of each athymic host. After 8 weeks, graft hosts were killed and their tumors removed, fixed and prepared for histological and immunohistochemical examination. Embryonic teratomas developed at high frequency from murine egg cylinders and from Day 11 and Day 12 porcine and Day 14 bovine embryos. Tumors were observed only infrequently from younger bovine and porcine blastocysts. Murine embryonic tumors were composed of numerous differentiated cell types of ectodermal, mesodermal and endodermal origins, but representation of the three embryonic germ layers was somewhat more restricted in bovine and porcine embryonic tumors. No undifferentiated stem cells were detected in tumors of any of the three species. These results demonstrate that teratomas will develop from bovine and porcine embryos when grafted to an immunocompromised host, but the presence of undifferentiated teratocarcinoma stem cells from these species has yet to be achieved. PMID- 9227926 TI - Laparoscopic surgery in infertile women with minimal or mild endometriosis. Canadian Collaborative Group on Endometriosis. AB - BACKGROUND: Minimal or mild endometriosis is frequently diagnosed in infertile women. It is often treated by resection or ablation of the lesions, but whether this improves fertility has not been established. We carried out a randomized, controlled trial to determine whether laparoscopic surgery enhanced fecundity in infertile women with minimal or mild endometriosis. METHODS: We studied 341 infertile women 20 to 39 years of age with minimal or mild endometriosis. During diagnostic laparoscopy the women were randomly assigned to undergo resection or ablation of visible endometriosis or diagnostic laparoscopy only. They were followed for 36 weeks after the laparoscopy or, for those who became pregnant during that interval, for up to 20 weeks of pregnancy. RESULTS: Among the 172 women who had resection or ablation of endometriosis, 50 became pregnant and had pregnancies that continued for 20 weeks or longer, as compared with 29 of the 169 women in the diagnostic-laparoscopy group (cumulative probabilities, 30.7 percent and 17.7 percent, respectively; P=0.006 by the log-rank test). The corresponding rates of fecundity were 4.7 and 2.4 per 100 person-months (rate ratio, 1.9; 95 percent confidence interval, 1.2 to 3.1). Fetal losses occurred in 20.6 percent of all the recognized pregnancies in the laparoscopic-surgery group and in 21.6 percent of all those in the diagnostic-laparoscopy group (P=0.91). Four minor operative complications (intestinal contusion, slight tear of the tubal serosa, difficult pneumoperitoneum, and vascular trauma) were reported (three in the surgery group and one in the control group). CONCLUSIONS: Laparoscopic resection or ablation of minimal and mild endometriosis enhances fecundity in infertile women. PMID- 9227927 TI - Interferon alfa-2b combined with cytarabine versus interferon alone in chronic myelogenous leukemia. French Chronic Myeloid Leukemia Study Group. AB - BACKGROUND: Treatment with interferon prolongs survival in chronic myelogenous leukemia. We conducted a clinical trial to assess the efficacy of treatment with a combination of interferon and cytarabine. METHODS: Previously untreated patients with chronic myelogenous leukemia were randomly assigned to receive either hydroxyurea (50 mg per kilogram of body weight per day) and interferon alfa-2b (5 million units per square meter of body-surface area per day), or hydroxyurea and interferon in the same dosages plus monthly courses of cytarabine (20 mg per square meter per day, for 10 days). The end points were overall survival, complete hematologic remission at 6 months, and major cytogenetic response (less than 35 percent Philadelphia chromosome-positive cells in the bone marrow) at 12 months. RESULTS: The trial was stopped when a sequential analysis showed a benefit of interferon and cytarabine. A significant improvement in survival was observed in the interferon-cytarabine group (360 patients) as compared with the interferon group (361 patients) (P=0.02; relative risk of death, 0.64; 95 percent confidence interval, 0.44 to 0.93). After three years, the survival rate was 85.7 percent with interferon and cytarabine and 79.1 percent with interferon alone. The rate of hematologic response was higher in the interferon-cytarabine group than in the interferon group (P=0.003). Major cytogenetic responses were observed 12 months after randomization in 126 of 311 patients treated with interferon and cytarabine (41 percent) and in 75 of 314 patients treated with interferon only (24 percent, P<0.001). CONCLUSIONS: The combination of interferon and cytarabine, as compared with interferon alone, increases the rate of major cytogenetic response and prolongs survival in patients with the chronic phase of chronic myelogenous leukemia. PMID- 9227928 TI - Plasma homocysteine levels and mortality in patients with coronary artery disease. AB - BACKGROUND: Elevated plasma homocysteine levels are a risk factor for coronary heart disease, but the prognostic value of homocysteine levels in patients with established coronary artery disease has not been defined. METHODS: We prospectively investigated the relation between plasma total homocysteine levels and mortality among 587 patients with angiographically confirmed coronary artery disease. At the time of angiography in 1991 or 1992, risk factors for coronary disease, including homocysteine levels, were evaluated. The majority of the patients subsequently underwent coronary-artery bypass grafting (318 patients) or percutaneous transluminal coronary angioplasty (120 patients); the remaining 149 were treated medically. RESULTS: After a median follow-up of 4.6 years, 64 patients (10.9 percent) had died. We found a strong, graded relation between plasma homocysteine levels and overall mortality. After four years, 3.8 percent of patients with homocysteine levels below 9 micromol per liter had died, as compared with 24.7 percent of those with homocysteine levels of 15 micromol per liter or higher. Homocysteine levels were only weakly related to the extent of coronary artery disease but were strongly related to the history with respect to myocardial infarction, the left ventricular ejection fraction, and the serum creatinine level. The relation of homocysteine levels to mortality remained strong after adjustment for these and other potential confounders. In an analysis in which the patients with homocysteine levels below 9 micromol per liter were used as the reference group, the mortality ratios were 1.9 for patients with homocysteine levels of 9.0 to 14.9 micromol per liter, 2.8 for those with levels of 15.0 to 19.9 micromol per liter, and 4.5 for those with levels of 20.0 micromol per liter or higher (P for trend=0.02). When death due to cardiovascular disease (which occurred in 50 patients) was used as the end point in the analysis, the relation between homocysteine levels and mortality was slightly strengthened. CONCLUSIONS: Plasma total homocysteine levels are a strong predictor of mortality in patients with angiographically confirmed coronary artery disease. PMID- 9227929 TI - Patient-to-patient transmission of hepatitis C virus during colonoscopy. PMID- 9227930 TI - Images in clinical medicine. Palate destruction by Aspergillus. PMID- 9227931 TI - Testicular germ-cell cancer. PMID- 9227932 TI - Fungal sinusitis. PMID- 9227933 TI - Case Records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 23-1997. A premature newborn infant with congenital ascites. PMID- 9227934 TI - Surgical treatment of minimal endometriosis. PMID- 9227936 TI - Communicating with patients who cannot read. PMID- 9227937 TI - Pressure ulcer prevalence, incidence, risk factors, and impact. AB - Pressure ulcers are a common problem among older adults in all health care settings. Prevalence and incidence estimates vary by setting, ulcer stage, and length of follow-up. Risk factors associated with increased pressure ulcer incidence have been identified. Activity or mobility limitation, incontinence, abnormalities in nutritional status, and altered consciousness are the most consistently reported risk factors for pressure ulcers. Pain, infectious complications, prolonged and expensive hospitalizations, persistent open ulcers, and increased risk of death are all associated with the development of pressure ulcers. The tremendous variability in pressure ulcer prevalence and incidence in health care settings suggests that opportunities exist to improve outcomes for persons at risk for and with pressure ulcers. PMID- 9227935 TI - Optimizing treatment for chronic myeloid leukemia. PMID- 9227938 TI - Strategies for preventing pressure ulcers. AB - The Agency for Health Care Policy and Research supported the development of guidelines for the prediction and prevention of pressure ulcers. Based on the best available scientific evidence, the guidelines recommend that individuals who are bed- or chair-bound should be assessed further for risk and receive care according to the risk factors. Appropriate actions include managing tissue loads, reducing exposure to moisture, managing incontinence, and assuring adequate nutrition. The guidelines are presented, research supporting the guidelines is summarized, and relevant articles published since the release of the guidelines are reviewed. PMID- 9227939 TI - Pressure ulcer assessment. AB - Pressure ulcer assessment requires quantification of multiple parameters of the ulcer and periulcer tissue. Clinical assessment should include ulcer history (including etiology, duration, and prior treatment), anatomic location, stage, size (including length, width, and depth measured in centimeters), sinus tracts, undermining, tunneling, exudate or drainage, necrotic tissue (slough and eschar), presence or absence of granulation tissue, and epithelialization. In addition, the ulcer borders can provide clues to healing potential. Intact skin surrounding the ulcer should be assessed for redness, warmth, induration or hardness, swelling, and any obvious signs of clinical infection. Pressure ulcer associated pain should be assessed prior to examination of the ulcer. PMID- 9227940 TI - Educational assessment and teaching of older clients with pressure ulcers. AB - The importance of education in preventing and treating patients with pressure ulcers is evident by the inclusion of recommendations for educational objectives in the clinical guidelines by the Agency for Health Care Policy and Research. Both caregiver and client should be assessed for knowledge of pressure ulcers. Physical changes in vision, hearing, and cognition that might affect educating older clients are described. Strategies suitable for teaching older clients about the prevention and treatment of pressure ulcers are given. PMID- 9227941 TI - The role of nutrition in prevention and healing of pressure ulcers. AB - Among the many risk factors for pressure ulcers, malnutrition is potentially reversible. This article examines the relationship of malnutrition to the prevention and healing of pressure ulcers. Evidence for nutrition in preventing and healing pressure ulcers is presented. Specific nutrients, including some amino acids, vitamins, and minerals, have been evaluated for their effects on wound healing. PMID- 9227942 TI - Pressure-relieving strategies for preventing and treating pressure sores. AB - Pressure-relieving strategies remain the foundation for the prevention and treatment of pressure sores. Although the published literature is inadequate, rational treatment decisions can be made if they are based on an understanding of the mechanisms by which pressure relief results with a particular device or strategy. This article reviews the theoretic and practical approaches to managing tissue loads so that rational choices of the most cost-effective strategies for preventing and treating pressure sores can be made when caring for patients. PMID- 9227943 TI - Pressure ulcers. Local wound care. AB - Local care of pressure ulcers includes wound cleansing, debridement, and dressings. Wound cleansing should remove loose debris and exudate but should not damage viable tissue. Saline irrigation is the standard. Debridement is often necessary for Stage III and IV pressure ulcers and can be performed autolytically, mechanically, enzymatically, or sharply. Prompt debridement is essential for infected wounds. Dressings should keep the wound bed continuously moist, should not be toxic to granulation tissue, and should keep the surrounding intact skin dry. Randomized, controlled clinical trials are necessary to define optimal local wound care further. PMID- 9227944 TI - Adjuvant therapy for ulcer care. AB - Adjuvant therapies, specifically electrotherapy, hyperbaric oxygen, ultrasound, and hydrotherapy, are considered increasingly for use with conventional local wound care to support healing of pressure ulcers. This article describes the characteristics of these modalities, their physiologic effects on the healing process, and the research to evaluate their efficacy. PMID- 9227945 TI - Pressure ulcers. Managing bacterial colonization and infection. AB - High levels of contamination are associated with delayed healing of pressure ulcers, and problems occur when surface contaminants invade the tissues and produce infection. This article addresses differentiating contamination from infection, identifying infection when it is present, and treating the patient with a contaminated or infected pressure ulcer. Systemic support for healing is discussed with a focus on oxygenation and nutrition. A comprehensive plan for the concomitant treatment of all factors contributing to pressure ulcer infection is recommended. PMID- 9227946 TI - Pressure ulcers. Assessment of healing. AB - Assessment of pressure ulcer healing involves observation during a complex series of cellular and molecular events that result in repair and restoration of skin integrity and function. Although the most important endpoint in healing is complete wound closure, it may not be a practical measurement in most settings. Assessment, therefore, often relies on measurement of partial healing, including changes in size and other wound characteristics over time. With no consensus about the best method for measurement of healing, it is not surprising that rates of healing have been described in only a few research studies. The establishment of a universal method for wound healing measurement is needed urgently. PMID- 9227947 TI - Operative repair of pressure ulcers. AB - Surgical management of pressure ulcers ranges from debridement and advancement flap closure for simple ulcers to sensate flaps, expanded flaps, free-tissue transfers, and fillet flaps for more complex ulcers and defects. Some pressure ulcers recur following surgery or conservative treatment, and the surgical options for management of these difficult recurrent ulcers are limited. The geriatric population offers an even more difficult problem as patients suffer invariably from underlying medical and systemic diseases that may affect surgery or the rehabilitation program. PMID- 9227948 TI - Quality assurance programs for pressure ulcers. AB - Traditional medical quality assurance programs are beginning to incorporate the principles of continuous quality improvement pioneered by Juran and Deming. Strategies for incorporating these principles into a long-term care facility are described, and two examples of successful implementation of continuous quality improvement programs for pressure ulcers are presented. PMID- 9227949 TI - Acquired motor neuron disorders. AB - The acquired motor neuron disorders are a heterogeneous group of conditions in which motor neuron degeneration or dysfunction produces the predominant manifestation of weakness, while the sensory system is clinically spared. The disorders most commonly seen in clinical practice are amyotrophic lateral sclerosis, late manifestations of poliomyelitis, and lower motor neuron syndromes, including motor neuropathy. Less often, acquired motor neuron disorders may complicate metabolic, toxic, or systemic disorders. The pathogenesis of most acquired motor neuron disorders is poorly understood, and treatment is mainly supportive; however clues to understanding the pathogenesis of amyotrophic lateral sclerosis are emerging, and new pharmacologic therapies are available. There is a growing sense of hope that combinations of drugs that are currently being tested may impact the survival of amyotrophic lateral sclerosis. PMID- 9227950 TI - Acquired peripheral neuropathy. AB - This article reviews the acquired causes of polyneuropathy other than diabetic and acute-onset neuropathies. The author gives a general method to simplify the diagnosis of chronic polyneuropathy. The acquired polyneuropathies are discussed under four main headings: metabolic disorders, toxic or deficiency states, infections, and immune-mediated. Recent advances in therapy are emphasized, and some illustrative case histories are provided. PMID- 9227951 TI - Acute peripheral neuropathy in adults. Guillain-Barre syndrome and related disorders. AB - Acute paralysis in adults has an extensive assortment of etiologies. Guillian Barre syndrome is the most common cause of acute neuropathy in adults. This review emphasizes pathophysiology, clinical features, differential diagnosis, and a practical approach to the laboratory work-up for patients with suspected Guillian-Barre syndrome. The current status of immunotherapy is reviewed. PMID- 9227952 TI - Diagnosis and management of common compression and entrapment neuropathies. AB - The most common focal neuropathies are carpal tunnel syndrome, ulnar neuropathy at the elbow, and peroneal neuropathy at the fibular head, but many other focal neuropathies, due to external compression or entrapment, may occur. Rational management depends on accurate localization; a thorough understanding of the basic anatomy, pathology, and pathophysiology helps in dealing with the vagaries of clinical presentation and electrodiagnostic evaluation. The differential diagnosis includes musculoskeletal conditions, plexopathies, radiculopathies, and occasionally, central nervous system dysfunction. Some focal neuropathies are an accentuation of a more generalized process, and a complex interplay of focal and diffuse pathology can arise. PMID- 9227953 TI - Diabetic neuropathy. AB - The most common form of diabetic neuropathy is chronic, distal symmetrical sensorimotor, or predominantly sensory neuropathy; the latter is invariably associated with some degree of autonomic dysfunction. There are, however, other neuropathic patterns in diabetes mellitus that are uncommon but are important to recognize, since they may mimic many other non-neurologic diseases. This article discusses a variety of forms of mononeuropathies and diabetic proximal motor neuropathy, commonly known as diabetic amyotropy. PMID- 9227954 TI - Acquired myasthenia gravis. AB - Myasthenia gravis, an antibody-mediated disorder of neuromuscular transmission that produces clinical weakness, may be ocular or generalized. Clinical diagnostic evaluation may be supplemented by electrophysiologic studies and antibody testing. Therapeutic options, including anticholinesterase inhibitors, immunosuppressive agents, plasmapheresis and thymectomy, are tailored for the individual patient. This article emphasizes the key aspects of the clinical evaluation, diagnosis, and therapy. PMID- 9227955 TI - Paraneoplastic neuromuscular syndromes. AB - Although paraneoplastic syndromes are rare, a number of well- defined, neuromuscular paraneoplastic syndromes have been described and their pathophysiology listed. Many different malignancies have been associated with these syndromes, but small-cell lung cancer is the most common. Features shared by these conditions include onset of the underlying malignancy, rapid progression, severe disability, and the potential for some improvement, owing to treatment of the cancer. This article discusses Lambert-Eaton myasthenic syndrome, motor neuron disorders, peripheral neuropathies, and disorders of continuous muscle fiber activity, such as Stiffman syndrome. PMID- 9227956 TI - Idiopathic inflammatory myopathies. AB - Dermatomyositis, polymyositis, and inclusion body myositis are the major categories of idiopathic inflammatory myopathy. These inflammatory myopathies are distinct clinically, histologically, and pathogenically. Features of dermatomyositis and polymyositis can overlap with those of other autoimmune connective tissue diseases. In this article, the authors review the characteristic features of these myopathies, update the recent developments in this area, and provide a framework for treatment. PMID- 9227957 TI - Myoglobinuria, malignant hyperthermia, neuroleptic malignant syndrome and serotonin syndrome. AB - This article presents an overview of the causes and manifestations of myoglobinuria and provides criteria for its diagnosis and management. The article also reviews neuroleptic malignant syndrome, malignant hyperthermia, and serotonin syndrome, all of which could cause rhabdomyolysis and myoglobinuria. PMID- 9227959 TI - Muscle pain, fatigue, and fasiculations. AB - This article discusses muscle pain, fatigue, and fasiculations. Muscle pain and fatigue are common problems in general medicine and in neurology, while fasiculations raise concern about a potentially ominous disease. The author reviews the conditions that cause pain and similar conditions arising from nonmuscular soft tissues. The article includes a general evaluation to be used for each of these clinical problems. PMID- 9227958 TI - Endocrine neuromyopathies. AB - The myopathies associated with endocrine disorders range in clinical presentation from the relatively nonspecific pattern of proximal muscle weakness of glucocorticoid excess states to specific presentations of contractions produced in tetany. All endocrine neuromyopathies emphasize the role of skeletal muscle in protein, carbohydrate, and electrolyte metabolism. Hormonal abnormalities tend to compromise muscle force generation by indirect effects on muscle function. The recognition and effective treatment of all these disorders require the identification of the underlying hormonal imbalances and awareness of general medical problems produced by the endocrine disorders. PMID- 9227960 TI - Toxic myopathies. AB - Toxic myopathies may occur with a variety of prescribed medications, illicit drug abuse, or other toxins. The article discusses an overview of some of the compounds that may cause myopathy, the clinical and laboratory features, histology, mechanisms of action, and potential risk factors of myopathy. The ability to recognize these syndromes is essential to avoid unnecessary tests and to avoid delay in treatment, especially in critically ill patients or patients with other neuromuscular diseases. PMID- 9227999 TI - Chlorine-36 in fossil rat urine: an archive of cosmogenic nuclide deposition during the past 40,000 years. AB - Knowledge of the production history of cosmogenic nuclides, which is needed for geological and archaeological dating, has been uncertain. Measurements of chlorine-36/chlorine (36Cl/Cl) ratios in fossil packrat middens from Nevada that are radiocarbon-dated between about 38 thousand years ago (ka) and the present showed that 36Cl/Cl ratios were higher by a factor of about 2 before approximately 11 ka. This raises the possibility that cosmogenic production rates just before the close of the Pleistocene were up to 50% higher than is suggested by carbon-14 calibration data. The discrepancy could be explained by addition of low-carbon-14 carbon dioxide to the atmosphere during that period, which would have depressed atmospheric radiocarbon activity. Alternatively, climatic effects on 36Cl deposition may have enhanced the 36Cl/Cl ratios. PMID- 9228004 TI - A di-acidic signal required for selective export from the endoplasmic reticulum. AB - Transport of membrane proteins between intracellular compartments requires specific sequences in the protein cytoplasmic domain to direct packaging into vesicle shuttles. A sequence that mediates export from the endoplasmic reticulum (ER) has proved elusive. A di-acidic signal (Asp-X-Glu, where X represents any amino acid) on the cytoplasmic tail of vesicular stomatitis virus glycoprotein (VSV-G) and other cargo molecules was required for efficient recruitment to vesicles mediating export from the ER in baby hamster kidney cells. The existence of such a signal provides evidence that export from the ER occurs through a selective mechanism. PMID- 9228005 TI - Bcl-2: prolonging life in a transgenic mouse model of familial amyotrophic lateral sclerosis. AB - Mutations in the gene encoding copper/zinc superoxide dismutase enzyme produce an animal model of familial amyotrophic lateral sclerosis (FALS), a fatal disorder characterized by paralysis. Overexpression of the proto-oncogene bcl-2 delayed onset of motor neuron disease and prolonged survival in transgenic mice expressing the FALS-linked mutation in which glycine is substituted by alanine at position 93. It did not, however, alter the duration of the disease. Overexpression of bcl-2 also attenuated the magnitude of spinal cord motor neuron degeneration in the FALS-transgenic mice. PMID- 9228006 TI - Prevention of vascular and neural dysfunction in diabetic rats by C-peptide. AB - C-peptide, a cleavage product from the processing of proinsulin to insulin, has been considered to possess little if any biological activity other than its participation in insulin synthesis. Injection of human C-peptide prevented or attenuated vascular and neural (electrophysiological) dysfunction and impaired Na+- and K+-dependent adenosine triphosphate activity in tissues of diabetic rats. Nonpolar amino acids in the midportion of the peptide were required for these biological effects. Synthetic reverse sequence (retro) and all-D-amino acid (enantio) C-peptides were equipotent to native C-peptide, which indicates that the effects of C-peptide on diabetic vascular and neural dysfunction were mediated by nonchiral interactions instead of stereospecific receptors or binding sites. PMID- 9228007 TI - Dual role of phosphatidylinositol-3,4,5-trisphosphate in the activation of protein kinase B. AB - Protein kinase B (PKB) is a proto-oncogene that is activated in signaling pathways initiated by phosphoinositide 3-kinase. Chromatographic separation of brain cytosol revealed a kinase activity that phosphorylated and activated PKB only in the presence of phosphatidylinositol-3,4,5-trisphosphate [PtdIns(3,4,5)P3]. Phosphorylation occurred exclusively on threonine-308, a residue implicated in activation of PKB in vivo. PtdIns(3,4,5)P3 was determined to have a dual role: Its binding to the pleckstrin homology domain of PKB was required to allow phosphorylation by the upstream kinase and it directly activated the upstream kinase. PMID- 9228008 TI - Transmission of hepatitis C by intrahepatic inoculation with transcribed RNA. AB - More than 1% of the world's population is chronically infected with hepatitis C virus (HCV). HCV infection can result in acute hepatitis, chronic hepatitis, and cirrhosis, which is strongly associated with development of hepatocellular carcinoma. Genetic studies of HCV replication have been hampered by lack of a bona fide infectious molecular clone. Full-length functional clones of HCV complementary DNA were constructed. RNA transcripts from the clones were found to be infectious and to cause disease in chimpanzees after direct intrahepatic inoculation. This work defines the structure of a functional HCV genome RNA and proves that HCV alone is sufficient to cause disease. PMID- 9228009 TI - Mitosis in living budding yeast: anaphase A but no metaphase plate. AB - Chromosome movements and spindle dynamics were visualized in living cells of the budding yeast Saccharomyces cerevisiae. Individual chromosomal loci were detected by expression of a protein fusion between green fluorescent protein (GFP) and the Lac repressor, which bound to an array of Lac operator binding sites integrated into the chromosome. Spindle microtubules were detected by expression of a protein fusion between GFP and Tub1, the major alpha tubulin. Spindle elongation and chromosome separation exhibited biphasic kinetics, and centromeres separated before telomeres. Budding yeast did not exhibit a conventional metaphase chromosome alignment but did show anaphase A, movement of the chromosomes to the poles. PMID- 9228011 TI - Superoxide anion radical (O2-.), superoxide dismutases, and related matters. PMID- 9228012 TI - Activation and involvement of p38 mitogen-activated protein kinase in glutamate induced apoptosis in rat cerebellar granule cells. AB - In the mammalian central nervous system glutamate is the major excitatory neurotransmitter and plays a crucial role in plasticity and toxicity of certain neural cells. We found that glutamate stimulated activation of p38 and stress activated protein kinase (SAPK, also known as c-Jun N-terminal kinase (JNK)), two subgroup members of the mitogen-activated protein kinase superfamily in matured cerebellar granule cells. The p38 activation was largely mediated by N-methyl-D aspartate receptors. Furthermore, we have revealed a novel signaling pathway, that is, Ca2+-mediated activation of p38 in glutamate-treated granule cells. The glutamate concentration effective for inducing apoptosis correlated with that for inducing p38 activation. SB203580, a specific inhibitor for p38, inhibited glutamate-induced apoptosis. Thus p38 might be involved in glutamate-induced apoptosis in cerebellar granule cells. PMID- 9228013 TI - Direct interaction of endothelial nitric-oxide synthase and caveolin-1 inhibits synthase activity. AB - Endothelial nitric-oxide synthase (eNOS) and caveolin-1 are associated within endothelial plasmalemmal caveolae. It is not known, however, whether eNOS and caveolin-1 interact directly or indirectly or whether the interaction affects eNOS activity. To answer these questions, we have cloned the bovine caveolin-1 cDNA and have investigated the eNOS-caveolin-1 interaction in an in vitro binding assay system using glutathione S-transferase (GST)-caveolin-1 fusion proteins and baculovirus-expressed bovine eNOS. We have also mapped the domains involved in the interaction using an in vivo yeast two-hybrid system. Results obtained using both in vitro and in vivo protein interaction assays show that both N- and C terminal cytosolic domains of caveolin-1 interact directly with the eNOS oxygenase domain. Interaction of eNOS with GST-caveolin-1 fusion proteins significantly inhibits enzyme catalytic activity. A synthetic peptide corresponding to caveolin-1 residues 82-101 also potently and reversibly inhibits eNOS activity by interfering with the interaction of the enzyme with Ca2+/calmodulin (CaM). Regulation of eNOS in endothelial cells, therefore, may involve not only positive allosteric regulation by Ca2+/CaM, but also negative allosteric regulation by caveolin-1. PMID- 9228014 TI - Expression cloning and characterization of a novel multispecific organic anion transporter. AB - Numerous drugs and endogenous compounds are efficiently excreted from the renal proximal tubule via carrier-mediated pathways. Transepithelial excretion of organic anions occurs via their accumulative transport from the blood into the proximal tubule cells across the basolateral membrane and subsequent secretion into the urine through the apical membrane. Here we report on the isolation of a novel complementary DNA from rat kidney that encodes a 551-amino acid residue protein (OAT1) with 12 putative membrane-spanning domains. When expressed in Xenopus laevis oocytes, OAT1 mediated sodium-independent para-aminohippurate (PAH) uptake (Km = 14.3 +/- 2.9 microM). The uptake rate of PAH was increased by the outwardly directed dicarboxylate gradient, consisting with the idea that OAT1 is an organic anion/dicarboxylate exchanger. OAT1 displayed remarkably wide substrate selectivity, covering endogenous substrates such as cyclic nucleotides, a prostaglandin and uric acid, and a variety of drugs with different structures (e.g. antibiotics, a nonsteroidal anti-inflammatory drug, diuretics, an antineoplastic drug, and a uricosuric drug). The Northern blot analysis and in situ hybridization revealed that OAT1 is exclusively expressed in the particular segment of the proximal tubule in the kidney. These data suggest that OAT1 is a multispecific organic anion transporter at the basolateral membrane of the proximal tubule. Isolation of OAT1 will facilitate elucidation of the molecular basis of drug kinetics and the development of new drugs lacking unwanted side effects. PMID- 9228015 TI - Zinc is a potent inhibitor of the apoptotic protease, caspase-3. A novel target for zinc in the inhibition of apoptosis. AB - The prevention of apoptosis by Zn2+ has generally been attributed to its inhibition of an endonuclease acting in the late phase of apoptosis. In this study we investigated the effect of Zn2+ on an earlier event in the apoptotic process, the proteolysis of the "death substrate" poly(ADP-ribose) polymerase (PARP). Pretreatment of intact Molt4 leukemia cells with micromolar concentrations of Zn2+ caused an inhibition of PARP proteolysis induced by the chemotherapeutic agent etoposide. Using a cell-free system consisting of purified bovine PARP as a substrate and an apoptotic extract or recombinant caspase-3 as the PARP protease, Zn2+ inhibited PARP proteolysis in the low micromolar range. To rule out an effect of Zn2+ on PARP, a protein with two zinc finger domains, we used recombinant caspase-3 and a chromogenic tetrapeptide substrate containing the caspase-3 cleavage site. In this system, Zn2+ inhibited caspase-3 with an IC50 of 0.1 microM. These results identify caspase-3 as a novel target of Zn2+ inhibition in apoptosis and suggest a regulatory role for Zn2+ in modulating the upstream apoptotic machinery. PMID- 9228016 TI - Cyclooxygenase-1 behaves as a delayed response gene in PC12 cells differentiated by nerve growth factor. AB - Treatment of PC12 cells with nerve growth factor (NGF) results in a differentiation program characterized by expression of immediate early and delayed response genes. In this program, morphological changes such as neurite extension are accompanied by phenotypic changes in enzyme expression, including an increased capacity for prostaglandin synthesis. Cyclooxygenase (COX), the enzyme responsible for prostanoid production, exists as two isoforms: constitutive COX-1 and inducible COX-2. We report that COX-1 behaves as a delayed response gene in PC12 cells exposed to NGF. Six hours following NGF treatment, COX-1 mRNA levels were elevated in PC12 cells, reaching nearly 5-fold above basal levels at 12 h. This increase was blocked by cycloheximide and was accompanied by concomitant increases in COX-1 protein and enzyme activity. COX-1 protein remained elevated for at least 10 days and localized to the cytoplasm and neurites of NGF-differentiated PC12 cells. Moreover, basic fibroblast growth factor, but not epidermal growth factor, caused similar increases in COX-1, which is consistent with expression characteristics of other delayed response genes in PC12 cells. This is the first example of neurotrophic factor regulation of cyclooxygenase and may have important implications for determination of the differentiated phenotype in PC12 cells. PMID- 9228017 TI - cDNA cloning of human retinoic acid-metabolizing enzyme (hP450RAI) identifies a novel family of cytochromes P450. AB - Retinoids, including all-trans-retinoic acid (RA) and its stereoisomer 9-cis-RA play important roles in regulating gene expression, through interactions with nuclear receptors, during embryonic development and in the maintenance of adult epithelial tissues (Chambon, P. (1995) Rec. Prog. Horm. Res. 50, 317-32; Mangelsdorf, D. J., and Evans, R. M. (1995) Cell 83, 841-850; Petkovich, M. (1992) Annu. Rev. Nutr. 12, 443-471). Evidence suggests that 4-hydroxylation of RA inside the target cell limits its biological activity and initiates a degradative process of RA leading to its eventual elimination. However, 18 hydroxylation and glucuronidation may also be important steps in this process. In this paper, we describe the cloning and characterization of the first mammalian retinoic acid-inducible retinoic acid-metabolizing cytochrome P450 (hP450RAI), which belongs to a novel class of cytochromes (CYP26). We demonstrate that hP450RAI is responsible for generation of several hydroxylated forms of RA, including 4-OH-RA, 4-oxo-RA, and 18-OH-RA. We also show that hP450RAI mRNA expression is highly induced by RA in certain human tumor cell lines and further show that RA-inducible RA metabolism may correlate with P450RAI expression. We conclude that this enzyme plays a key role in RA metabolism, functioning in a feedback loop where RA levels are controlled in an autoregulatory manner. PMID- 9228018 TI - FLAME-1, a novel FADD-like anti-apoptotic molecule that regulates Fas/TNFR1 induced apoptosis. AB - We identified and cloned a novel human protein that contains FADD/Mort1 death effector domain homology regions, designated FLAME-1. FLAME-1, although most similar in structure to Mch4 and Mch5, does not possess caspase activity but can interact specifically with FADD, Mch4, and Mch5. Interestingly, FLAME-1 is recruited to the Fas receptor complex and can abrogate Fas/TNFR-induced apoptosis upon expression in FasL/tumor necrosis factor-sensitive MCF-7 cells, possibly by acting as a dominant-negative inhibitor. These findings identify a novel endogenous control point that regulates Fas/TNFR1-mediated apoptosis. PMID- 9228020 TI - Combinatorial mutations of lac repressor. Stability of monomer-monomer interface is increased by apolar substitution at position 84. AB - To examine the monomer-monomer subunit interface in the lac repressor, a mutation that generates dimeric protein (deletion of C-terminal amino acids to disrupt the dimer-dimer interface) has been combined with amino acid substitutions that alter the monomer-monomer interface (substitution at Lys84 or Tyr282). Dimeric proteins with significantly increased stability to urea denaturation were formed by the introduction of the apolar amino acids Ala or Leu in lieu of Lys84 in concert with the deletion of 11 C-terminal amino acids. K84A/-11 deletion protein retained wild-type affinity for operator DNA, while K84L/-11 deletion protein displayed operator affinity similar to its parent tetramer. To assess further the influence of monomer-monomer interface stability on assembly and DNA binding, triple mutants were generated with Y282D, an alteration that disrupts assembly completely in the wild-type background. The triple mutants were dimeric, but they exhibited diminished dimer stability to urea denaturation and decreased operator affinity compared with the double mutations. These results demonstrate directly the stabilizing influence of apolar substitution at position 84 on the monomer monomer interface. PMID- 9228019 TI - Constitutive activation of the beta2 adrenergic receptor alters the orientation of its sixth membrane-spanning segment. AB - The binding site of the beta2 adrenergic receptor, like that of other homologous G-protein-coupled receptors, is contained within a water-accessible crevice formed among its seven membrane-spanning segments. Methanethiosulfonate ethylammonium (MTSEA), a charged, hydrophilic, lipophobic, sulfhydryl-specific reagent, had no effect on the binding of agonist or antagonist to wild-type beta2 receptor expressed in HEK 293 cells. This suggested that no endogenous cysteines are accessible in the binding site crevice. In contrast, in a constitutively active beta2 receptor, MTSEA significantly inhibited antagonist binding, and isoproterenol slowed the rate of reaction of MTSEA. This implies that at least one endogenous cysteine becomes accessible in the binding site crevice of the constitutively active beta2 receptor. Cys-285, in the sixth membrane-spanning segment, is responsible for the inhibitory effect of MTSEA on ligand binding to the constitutively active mutant. The acquired accessibility of Cys-285 in the constitutively active mutant may result from a rotation and/or tilting of the sixth membrane-spanning segment associated with activation of the receptor. This rearrangement could bring Cys-285 to the margin of the binding site crevice where it becomes accessible to MTSEA. PMID- 9228021 TI - X-ray structure of human class IV sigmasigma alcohol dehydrogenase. Structural basis for substrate specificity. AB - The structural determinants of substrate recognition in the human class IV, or sigmasigma, alcohol dehydrogenase (ADH) isoenzyme were examined through x-ray crystallography and site-directed mutagenesis. The crystal structure of sigmasigma ADH complexed with NAD+ and acetate was solved to 3-A resolution. The human beta1beta1 and sigmasigma ADH isoenzymes share 69% sequence identity and exhibit dramatically different kinetic properties. Differences in the amino acids at positions 57, 116, 141, 309, and 317 create a different topology within the sigmasigma substrate-binding pocket, relative to the beta1beta1 isoenzyme. The nicotinamide ring of the NAD(H) molecule, in the sigmasigma structure, appears to be twisted relative to its position in the beta1beta1 isoenzyme. In conjunction with movements of Thr-48 and Phe-93, this twist widens the substrate pocket in the vicinity of the catalytic zinc and may contribute to this isoenzyme's high Km for small substrates. The presence of Met-57, Met-141, and Phe-309 narrow the middle region of the sigmasigma substrate pocket and may explain the substantially decreased Km values with increased chain length of substrates in sigmasigma ADH. The kinetic properties of a mutant sigmasigma enzyme (sigma309L317A) suggest that widening the middle region of the substrate pocket increases Km by weakening the interactions between the enzyme and smaller substrates while not affecting the binding of longer alcohols, such as hexanol and retinol. PMID- 9228023 TI - Metabolic fate of glucose in purified islet cells. Glucose-regulated anaplerosis in beta cells. AB - Previous studies in rat islets have suggested that anaplerosis plays an important role in the regulation of pancreatic beta cell function and growth. However, the relative contribution of islet beta cells versus non-beta cells to glucose regulated anaplerosis is not known. Furthermore, the fate of glucose carbon entering the Krebs cycle of islet cells remains to be determined. The present study has examined the anaplerosis of glucose carbon in purified rat beta cells using specific 14C-labeled glucose tracers. Between 5 and 20 mM glucose, the oxidative production of CO2 from [3,4-14C]glucose represented close to 100% of the total glucose utilization by the cells. Anaplerosis, quantified as the difference between 14CO2 production from [3,4-14C]glucose and [6-14C]glucose, was strongly influenced by glucose, particularly between 5 and 10 mM. The dose dependence of glucose-induced insulin secretion correlated with the accumulation of citrate and malate in beta(INS-1) cells. All glucose carbon that was not oxidized to CO2 was recovered from the cells after extraction in trichloroacetic acid. This indirectly indicates that lactate output is minimal in beta cells. From the effect of cycloheximide upon the incorporation of 14C-glucose into the acid-precipitable fraction, it could be calculated that 25% of glucose carbon entering the Krebs cycle via anaplerosis is channeled into protein synthesis. In contrast, non-beta cells (approximately 80% glucagon-producing alpha cells) exhibited rates of glucose oxidation that were (1)/(3) to (1)/(6) those of the total glucose utilization and no detectable anaplerosis from glucose carbon. This difference between the two cell types was associated with a 7-fold higher expression of the anaplerotic enzyme pyruvate carboxylase in beta cells, as well as a 4-fold lower ratio of lactate dehydrogenase to FAD-linked glycerol phosphate dehydrogenase in beta cells versus alpha cells. Finally, glucose caused a dose dependent suppression of the activity of the pentose phosphate pathway in beta cells. In conclusion, rat beta cells metabolize glucose essentially via aerobic glycolysis, whereas glycolysis in alpha cells is largely anaerobic. The results support the view that anaplerosis is an essential pathway implicated in beta cell activation by glucose. PMID- 9228022 TI - Mutational analysis of the murine granzyme B gene promoter in primary T cells and a T cell clone. AB - The granzyme B gene is induced in cytotoxic T lymphocytes in response to antigenic stimulation. Previous studies have identified several distinct regions in the granzyme B promoter which may be important in either the induction or the T cell specificity of the gene. These regions contain the canonical transcription factor binding sites AP1, cyclic AMP-responsive element (CRE), Ikaros, and core binding factor (CBF/PEBP2). Each protein binding site was disrupted by site directed mutagenesis to investigate its role in granzyme B promoter function. Mutations were introduced alone, or in various combinations, within the context of a 243-base pair promoter fragment known to confer high levels of reporter gene expression. Transfection assays revealed that all of the single binding site mutant promoters were capable of sustaining moderate to high levels of transcriptional activity in primary activated T lymphocytes, whereas certain mutants were more impeded in a T cell clone. A quadruple mutant promoter, with only the CRE binding site intact, showed background expression levels. This drop in expression was found to be mostly due to mutations in AP1 and the 3' CBF binding sites. Their close proximity and requirement in promoter function suggest an important role for protein-protein interaction between these two factors. PMID- 9228024 TI - Deletion of two exons from the Lymnaea stagnalis beta1-->4-N acetylglucosaminyltransferase gene elevates the kinetic efficiency of the encoded enzyme for both UDP-sugar donor and acceptor substrates. AB - Lymnaea stagnalis UDP-GlcNAc:GlcNAcbeta-R beta1-->4-N acetylglucosaminyltransferase (beta4-GlcNAcT) is an enzyme with structural similarity to mammalian UDP-Gal:GlcNAcbeta-R beta1-->4-galactosyltransferase (beta4-GalT). Here, we report that also the exon organization of the genes encoding these enzymes is very similar. The beta4-GlcNAcT gene (12.5 kilobase pairs, spanning 10 exons) contains four exons, encompassing sequences that are absent in the beta4-GalT gene. Two of these exons (exons 7 and 8) show a high sequence similarity to part of the preceding exon (exon 6), suggesting that they have originated by exon duplication. The exon in the beta4-GalT gene, corresponding to beta4-GlcNAcT exon 6, encodes a region that has been proposed to be involved in the binding of UDP-Gal. The question therefore arose, whether the repeating sequences encoded by exon 7 and 8 of the beta4-GlcNAcT gene would determine the specificity of the enzyme for UDP-GlcNAc, or for the less preferred UDP-GalNAc. It was found that deletion of only the sequence encoded by exon 8 resulted in a completely inactive enzyme. By contrast, deletion of the amino acid residues encoded by exons 7 and 8 resulted in an enzyme with an elevated kinetic efficiency for both UDP-sugar donors, as well as for its acceptor substrates. These results suggest that at least part of the donor and acceptor binding domains of the beta4-GlcNAcT are structurally linked and that the region encompassing the insertion contributes to acceptor recognition as well as to UDP sugar binding and specificity. PMID- 9228025 TI - A dominant negative to activation protein-1 (AP1) that abolishes DNA binding and inhibits oncogenesis. AB - We describe a dominant negative (DN) to activation protein-1 (AP1) that inhibits DNA binding in an equimolar competition. AP1 is a heterodimer of the oncogenes Fos and Jun, members of the bZIP family of transcription factors. The DN, termed A-Fos, consists of a newly designed acidic amphipathic protein sequence appended onto the N-terminus of the Fos leucine zipper, replacing the normal basic region critical for DNA binding. The acidic extension and the Jun basic region form a heterodimeric coiled coil structure that stabilizes the complex over 3000-fold and prevents the basic region of Jun from binding to DNA. Gel shift assays indicate that A-Fos can inactivate the DNA binding of a Fos:Jun heterodimer in an equimolar competition. Transient transfection assays indicate that A-Fos inhibits Jun-dependent transactivation. Both the acidic extension and the Fos leucine zipper are critical for this inhibition. Expression of A-Fos in mouse fibroblasts inhibits focus formation more than colony formation, reflecting the ability of A Fos to interfere with the AP1 biological functions in mammalian cells. This reagent is more potent than a deletion of either the Fos or Jun transactivation domain, which has been used previously as a dominant negative to AP1 activity. PMID- 9228026 TI - Complex inhibition of OmpF and OmpC bacterial porins by polyamines. AB - The effects of four polyamines (putrescine, cadaverine, spermidine, and spermine) on the activity of bacterial porins OmpC and OmpF were investigated by electrophysiology. Membrane vesicles made from the outer membrane of Escherichia coli strains expressing only OmpC or OmpF were reconstituted into liposomes probed by patch clamp. The channel activity was recorded in control solutions and in the presence of increasing concentrations of a specific polyamine. In all cases, concentration- and voltage-dependent inhibitory effects were observed. They include both the suppression of channel openings and the enhancement of channel closures as well as the promotion of blocked or inactivated states. OmpF and OmpC, although highly homologous, have distinct sensitivities to modulation, especially by spermine. This compound inhibits OmpF in the nanomolar range, which is in agreement with its potency on eukaryotic channels. Putrescine was the least effective (upper millimolar range) and also had inhibitory effects qualitatively distinct from those exerted by the other polyamines. The compounds appear to bind to at least two distinct binding sites, one of which resides within the pore. The potencies to this site are lower when the polyamines are applied from the extracellular side than from the periplasmic side, suggesting an asymmetric binding site. PMID- 9228027 TI - Activation of R-Ras by Ras-guanine nucleotide-releasing factor. AB - Ras-GRF/CDC25(Mm), mSos, and C3G have been identified as guanine nucleotide releasing factors for Ras family proteins. We investigated in this study the guanine nucleotide-releasing activities of Ras-GRF, mSos, and C3G toward R-Ras, which shows high sequence similarity to Ras. Ras-GRF markedly stimulated the dissociation of GDP from R-Ras, and C3G also promoted the release of R-Ras-bound GDP. Under the same conditions, mSos little affected the reaction. When Ras-GRF and R-Ras were coexpressed in COS7 cells, the remarkable accumulation of the active GTP-bound form of R-Ras was observed. C3G also increased active R-Ras in COS7 cells, while mSos did not give any effect. These results indicated that Ras GRF and C3G could activate R-Ras. Furthermore, the activation of R-Ras by Ras-GRF was enhanced when cells were treated with ionomycin, which is known to increase the intracellular calcium concentration. The examination of tissue distribution of R-Ras, Ras-GRF, and mSos by the reverse transcription-polymerase chain reaction revealed that Ras-GRF was expressed only in brain and testis, whereas R Ras, C3G, and mSos were expressed rather ubiquitously. These findings raise the possibility that R-Ras is activated by Ras-GRF in brain and testis, and by C3G in other tissues, respectively. PMID- 9228028 TI - ATP-binding properties of human Hsp90. AB - Hsp90 is one of the most abundant proteins in the cytosol of eukaryotic cells. Under physiological conditions Hsp90 has been shown to play a major role in several specific signaling pathways, including maturation of various kinases and maintenance of steroid receptors in an activable state. It is well established that the level of Hsp90 increases severalfold under stress conditions, and it has been shown that the chaperone function of Hsp90 is ATP-independent. Although yeast Hsp90 does not bind ATP, as determined by a number of methods monitoring tight binding, ATP-dependent functions of Hsp90 in the presence of co-factors and elevated temperatures are still under discussion. Here, we have reinvestigated ATP-binding properties and ATPase activity of human Hsp90 under various conditions. We show that human Hsp90 does not bind ATP tightly and does not exhibit detectable ATPase activity. However, using electron spin resonance spectroscopy, weak binding of spin-labeled ATP analogues with half-maximal binding at 400 microM ATP was detected. The functional significance of this weak interaction remains enigmatic. PMID- 9228029 TI - Mutation of a highly conserved arginine in motif IV of Escherichia coli DNA helicase II results in an ATP-binding defect. AB - A site-directed mutation in motif IV of Escherichia coli DNA helicase II (UvrD) was generated to examine the functional significance of this region. The highly conserved arginine at position 284 was replaced with alanine to construct UvrD R284A. The ability of the mutant allele to function in methyl-directed mismatch repair and UvrABC-mediated nucleotide excision repair was examined by genetic complementation assays. The R284A substitution abolished function in both DNA repair pathways. To identify the biochemical defects responsible for the loss of biological function, UvrD-R284A was purified to apparent homogeneity, and its biochemical properties were compared with wild-type UvrD. UvrD-R284A failed to unwind a 92-base pair duplex region and was severely compromised in unwinding a 20-base pair duplex region. The Km of UvrD-R284A for ATP was significantly greater than 3 mM compared with 80 microM for UvrD. A large decrease in ATP binding was confirmed using a nitrocellulose filter binding assay. These data suggested that the R284A mutation severely reduced the affinity of helicase II for ATP. The reduced unwinding activity and loss of biological function of UvrD R284A was probably the result of decreased affinity for ATP. These results implicate motif IV of superfamily I helicases in nucleotide binding and represent the first characterization of a helicase mutation outside motifs I and II that severely impacted the Km for ATP. PMID- 9228030 TI - Engineering of glycerol-stimulated insulin secretion in islet beta cells. Differential metabolic fates of glucose and glycerol provide insight into mechanisms of stimulus-secretion coupling. AB - Insulin secretion from beta cells in the islets of Langerhans can be stimulated by a number of metabolic fuels, including glucose and glyceraldehyde, and is thought to be mediated by metabolism of the secretagogues and an attendant increase in the ATP:ADP ratio. Curiously, glycerol fails to stimulate insulin secretion, even though it has been reported that islets contain abundant glycerol kinase activity and oxidize glycerol efficiently. We have reinvestigated this point and find that rat islets and the well differentiated insulinoma cell line INS-1 contain negligible glycerol kinase activity. A recombinant adenovirus containing the bacterial glycerol kinase gene (AdCMV-GlpK) was constructed and used to express the enzyme in islets and INS-1 cells, resulting in insulin secretion in response to glycerol. In AdCMV-GlpK-treated INS-1 cells a greater proportion of glycerol is converted to lactate and a lesser proportion is oxidized compared with glucose. The two fuels are equally potent as insulin secretagogues, despite the fact that oxidation of glycerol at its maximally effective dose (2-5 mM) occurs at a rate that is similar to the rate of glucose oxidation at its basal, nonstimulatory concentration (3 mM). We also investigated the possibility that glycerol may signal via expansion of the glycerol phosphate pool to allow enhanced fatty acid esterification and formation of complex lipids. Addition of Triacsin-C, an inhibitor of long-chain acyl-CoA synthetase, to AdCMV GlpK-treated INS-1 cells did not inhibit glycerol-stimulated insulin secretion despite the fact that it blocked glycerol incorporation into cellular lipids. We conclude from these studies that glycerol kinase expression is sufficient to activate glycerol signaling in beta cells, showing that the failure of normal islets to respond to this substrate is due to a lack of this enzyme activity. Further, our studies show that glycerol signaling is not linked to esterification or oxidation of the substrate, but is likely mediated by its metabolism in the glycerol phosphate shuttle and/or the distal portion of the glycolytic pathway, either of which can lead to production of ATP and an increased ATP:ADP ratio. PMID- 9228031 TI - Mice deficient in lysosomal acid phosphatase develop lysosomal storage in the kidney and central nervous system. AB - Lysosomal acid phosphatase (LAP) is a tartrate-sensitive enzyme with ubiquitous expression. Neither the physiological substrates nor the functional significance is known. Mice with a deficiency of LAP generated by targeted disruption of the LAP gene are fertile and develop normally. Microscopic examination of various peripheral organs revealed progredient lysosomal storage in podocytes and tubular epithelial cells of the kidney, with regionally different ultrastructural appearance of the stored material. Within the central nervous system, lysosomal storage was detected to a regionally different extent in microglia, ependymal cells, and astroglia concomitant with the development of a progressive astrogliosis and microglial activation. Whereas behavioral and neuromotor analyses were unable to distinguish between control and deficient mice, approximately 7% of the deficient animals developed generalized seizures. From the age of 6 months onward, conspicuous alterations of bone structure became apparent, resulting in a kyphoscoliotic malformation of the lower thoracic vertebral column. We conclude from these findings that LAP has a unique function in only a subset of cells, where its deficiency causes the storage of a heterogeneously appearing material in lysosomes. The causal relationship of the enzyme defect to the clinical manifestations remains to be determined. PMID- 9228033 TI - Interaction of apolipoprotein J-amyloid beta-peptide complex with low density lipoprotein receptor-related protein-2/megalin. A mechanism to prevent pathological accumulation of amyloid beta-peptide. AB - Apolipoprotein J (apoJ) has been shown to be the predominant amyloid beta-peptide (Abeta)-binding protein in cerebrospinal fluid. We have previously demonstrated that the endocytic receptor low density lipoprotein receptor-related protein 2/megalin (LRP-2), which is expressed by choroid plexus epithelium and ependymal cells lining the brain ventricles and neural tube, binds and mediates cellular uptake of apoJ (Kounnas, M. Z., Loukinova, E. B., Stefansson, S., Harmony, J. A., Brewer, B., Strickland, D. K., and Argraves, W. S. (1995) J. Biol. Chem. 270, 13070-13075). In the present study, we evaluated the ability of apoJ to mediate binding of Abeta1-40-apoJ complex to LRP-2 in vitro. Immunoblot analysis showed that incubation of apoJ with Abeta1-40 resulted in the formation of Abeta1-40 apoJ complex and the inhibition of the formation of Abeta1-40 aggregates. Using an enzyme-linked immunosorbent assay, an estimated dissociation constant (Kd) of 4.8 nM was derived for the interaction between Abeta1-40 and apoJ. Enzyme-linked immunosorbent assay was also used to study the interaction of the Abeta1-40-apoJ complex with LRP-2. The results showed that Abeta alone did not bind directly to LRP-2; however, when Abeta1-40 was combined with apoJ to form a complex, binding to LRP-2 took place. The binding interaction could be blocked by inclusion of the receptor-associated protein, an antagonist of apoJ binding to LRP-2. When LRP-2 expressing cells were given 125I-Abeta1-40, cellular uptake of the radiolabeled peptide was promoted by co-incubation with apoJ. When the cells were provided purified 125I-Abeta1-40-apoJ complex, the complex was internalized and degraded, and both processes were inhibited with polyclonal LRP-2 antibodies. Furthermore, chloroquine treatment inhibited the cellular degradation of the complex. The data indicate that apoJ facilitates Abeta1-40 binding to LRP-2 and that the receptor mediates cellular clearance of Abeta1-40-apoJ complex leading to lysosomal degradation of Abeta1-40. The findings support the possibility that LRP-2 can act in vivo to mediate clearance of the complex from biological fluids such as cerebrospinal fluid and thereby play a role in the regulation of Abeta accumulation. PMID- 9228032 TI - Characterization of the osteoclast ruffled border chloride channel and its role in bone resorption. AB - Bone resorption by osteoclasts requires massive transcellular acid transport, which is accomplished by the parallel action of a V-type proton pump and a chloride channel in the osteoclast ruffled border. We have studied the molecular basis for the appearance of acid transport as avian bone marrow mononuclear cells acquire a bone resorptive phenotype in vitro. We demonstrate a critical role for regulated expression of a ruffled border chloride channel as the cells become competent to resorb bone. Molecular characterization of the chloride channel shows that it is related to the renal microsomal chloride channel, p64. In planar bilayers, the ruffled border channel is a stilbene sulfonate-inhibitable, outwardly rectifying chloride channel. A mechanism by which outward rectification of the single channel chloride current could allow efficient regulation of acidification by the channel is discussed. PMID- 9228034 TI - Identification of common ligand binding determinants of the insulin and insulin like growth factor 1 receptors. Insights into mechanisms of ligand binding. AB - Insulin and insulin-like growth factor 1 (IGF-1) are peptides that share nearly 50% sequence homology. However, although their cognate receptors also exhibit significant overall sequence homology, the affinity of each peptide for the non cognate receptor is 2-3 orders of magnitude lower than for the cognate receptor. The molecular basis for this discrimination is unclear, as are the molecular mechanisms underlying ligand binding. We have recently identified a major ligand binding site of the insulin receptor by alanine scannning mutagenesis. These studies revealed that a number of amino acids critical for insulin binding are conserved in the IGF-1 receptor, suggesting that they may play a role in ligand binding. We therefore performed alanine mutagenesis of these amino acids to determine whether this is the case. cDNAs encoding alanine-substituted secreted recombinant IGF-1 receptors were expressed in 293 EBNA cells, and the ligand binding properties of the expressed proteins were evaluated. Mutation of Phe701 resulted in a receptor with undetectable IGF-1 binding; alanine substitution of the corresponding amino acid of the insulin receptor, Phe714, produces a 140-fold reduction in affinity for insulin. Mutation of Asp8, Asn11, Phe58, Phe692, Glu693, His697, and Asn698 produces a 3.5-6-fold reduction in affinity for IGF-1. In contrast, alanine mutation of the corresponding amino acids of the insulin receptor with the exception of Asp12 produces reductions in affinity that are 50 fold or greater. The affinity of insulin for these mutants relative to wild type receptor was similar to that of their relative affinity for IGF-1 with two exceptions; the IC50 values for insulin binding to the mutants of Arg10, which has normal affinity for IGF-1, and His697, which has a 6-fold reduction in affinity for IGF-1, were both at least 2 orders of magnitude greater than for wild type receptor. The Kd values for insulin of the corresponding alanine mutants of the insulin receptor, Arg14 and His710, are 2-3 orders of magnitude greater than for wild type receptor. However, in contrast, the relative affinity of des(25-30)[PheB25 alpha-carboxamide]insulin for these IGF-1 receptor mutants is reduced only 4- and 50-fold, respectively. PMID- 9228035 TI - Mitosis-specific negative regulation of epidermal growth factor receptor, triggered by a decrease in ligand binding and dimerization, can be overcome by overexpression of receptor. AB - The function of epidermal growth factor receptor (EGFR) was found to be negatively regulated in M phase in which it showed less phosphotyrosine content and reduced intrinsic kinase activity accompanied by retarded electrophoretic mobility owing to total hyperphosphorylation. Ligand-induced autophosphorylation and downstream signaling of EGFR were tightly suppressed in M phase due to a decrease in ligand binding affinity and the inability of epidermal growth factor (EGF) to induce receptor dimerization. There was no change in the number of surface-exposed EGF receptors between G0/G1 and M phases of the cell cycle. Hyperphosphorylation (due to serine and/or threonine phosphorylation) correlates with the unresponsiveness of cells to EGF-mediated stimulation of tyrosine phosphorylation in cells that express the normal or basal level of EGFR. This M phase-specific negative regulation was overcome by overexpression of EGFR, which was responsive to ligand throughout the cell cycle and revealed ligand-induced signaling in the M phase. These findings indicate that EGFR does not respond to ligand stimulation in M phase and suggest that a negative regulation of ligand receptor interactions in M phase may control the normal function of receptor tyrosine kinase and that receptor overexpression will disrupt this cell cycle dependent regulation of receptor tyrosine kinases. PMID- 9228036 TI - Both overlapping and distinct signaling pathways for somatostatin receptor subtypes SSTR1 and SSTR2 in pituitary cells. AB - To elucidate the signaling events mediated by specific somatostatin receptor (SSTR) subtypes, we expressed SSTR1 and SSTR2 individually in rat pituitary GH12C1 and F4C1 cells, which lack endogenous somatostatin receptors. In transfected GH12C1 cells, both SSTR1 and SSTR2 coupled to inhibition of Ca2+ influx and hyperpolarization of membrane potential via a pertussis toxin (PTx) sensitive mechanism. These effects reflected modulation of ion channel activities which are important for regulation of hormone secretion. Somatostatin analogs MK678 and CH275 acted as subtype selective agonists as expected. In transfected F4C1 cells, both SSTR1 and SSTR2 mediated somatostatin-induced inhibition of adenylyl cyclase via a PTx-sensitive pathway. In addition, activation of SSTR2 in F4C1 cells, but not SSTR1, stimulated phospholipase C (PLC) activity and an increase in [Ca2+]i due to release of Ca2+ from intracellular stores. Unlike adenylyl cyclase inhibition, the PLC-mediated response was only partially sensitive to PTx. To determine the structural determinants in SSTR2 necessary for activation of PLC, we constructed chimeric receptors in which domains of SSTR2 were introduced into SSTR1. Chimeric receptors containing only the third intracellular loop, or all three intracellular loops from SSTR2, mediated inhibition of adenylyl cyclase, but failed to stimulate PLC activity as did wild type SSTR2. Furthermore, the C-terminal tail of SSTR2 was not required for coupling to PLC. Thus, by expressing individual somatostatin receptor subtypes in pituitary cells, we have identified both overlapping and distinct signaling pathways for SSTR1 and SSTR2, and have shown that sequences other than simply the intracellular domains are required for SSTR2 to couple to the PLC signaling pathway. PMID- 9228037 TI - Identification of an 8-lipoxygenase pathway in nervous tissue of Aplysia californica. AB - Arachidonic acid is converted to (8R)-hydroperoxyeicosa-5,9,11, 14-tetraenoic acid (8-HPETE) during incubations with homogenates of the central nervous system of the marine mollusc, Aplysia californica. 8-HPETE can be reduced to the corresponding hydroxy acid or be enzymatically converted to a newly identified metabolite, 8-ketoeicosa-5,9,11,14-tetraenoic acid (8-KETE). These metabolites were identified by high performance liquid chromatography, UV absorbance, and gas chromatography/mass spectrometry. Stereochemical analysis of the products demonstrate that the neuronal enzyme is an (8R)-lipoxygenase. Previously we have shown that the neurotransmitters, histamine and Phe-Met-Arg-Phe-amide, activate 12-lipoxygenase metabolism in isolated identified Aplysia neurons. We now show that acetylcholine activates the (8R)-lipoxygenase pathway within intact nerve cells. Thus, both (12S)- and (8R)-lipoxygenase co-exist in intact Aplysia nervous tissue but are differentially activated by several neurotransmitters. The precise physiological role of the 8-lipoxygenase products is currently under investigation, but by analogy to the well-described 12-lipoxygenase pathway, we suggest that (8R)-HPETE and 8-KETE may serve as second messengers in Aplysia cholinoceptive neurons. PMID- 9228039 TI - Conformational stabilities of Escherichia coli RNase HI variants with a series of amino acid substitutions at a cavity within the hydrophobic core. AB - Escherichia coli ribonuclease HI has a cavity within the hydrophobic core. Two core residues, Ala52 and Val74, resided at both ends of this cavity. We have constructed a series of single mutant proteins at Ala52, and double mutant proteins, in which Ala52 was replaced by Gly, Val, Ile, Leu, or Phe, and Val74 was replaced by Ala or Leu. All of these mutant proteins, except for A52W, A52R, and A52G/V74A, were overproduced and purified. Measurement of the thermal denaturations of the proteins at pH 3.2 by CD suggests that the cavity is large enough to accommodate three methyl or methylene groups without creating serious strains. A correlation was observed between the protein stability and the hydrophobicity of the substituted residue. As a result, a number of the mutant proteins were more stable than the wild-type protein. The stabilities of the mutant proteins with charged or extremely bulky residues at the cavity were lower than those expected from the hydrophobicities of the substituted residues, suggesting that considerable strains are created at the mutation sites in these mutant proteins. However, examination of the far- and near-UV CD spectra and the enzymatic activities suggest that all of the mutant proteins have structures similar to that of the wild-type protein. These results suggest that the cavity in the hydrophobic core of E. coli RNase HI is conformationally fairly stable. This may be the reason why the cavity-filling mutations effectively increase the thermal stability of this protein. PMID- 9228038 TI - Lipopolysaccharide (LPS)-binding proteins BPI and LBP form different types of complexes with LPS. AB - Lipopolysaccharide (LPS)-binding protein (LBP) and bactericidal/permeability increasing protein (BPI) are closely related LPS-binding proteins whose binding to LPS has markedly different functional consequences. To gain better insight into the possible basis of these functional differences, the physical properties of LBP-LPS and BPI-LPS complexes have been compared in this study by sedimentation, light scattering, and fluorescence analyses. These studies reveal dramatic differences in the physical properties of LPS complexed to LBP versus BPI. They suggest that of the two proteins, only LBP can disperse LPS aggegates. However, BPI can enhance both the sedimentation velocity and apparent size of LPS aggregates while inhibiting LPS-LBP binding even at very low (1:40 to 1:20) BPI:LPS molar ratios. PMID- 9228040 TI - The function of steroid hormone receptors is inhibited by the hsp90-specific compound geldanamycin. AB - The ansamycin antibiotic geldanamycin, which specifically interacts with the heat shock protein hsp90, was used to study the function of hsp90 in steroid hormone receptors. We observed inhibition of glucocorticoid-specific gene induction in several responsive cell systems. Hormone binding abilities of receptors for glucocorticoid, progestin, androgen, and estrogen were inhibited upon exposing intact cells to geldanamycin. Inhibition was only seen when geldanamycin was applied to cell cultures under growth conditions or was present during in vitro synthesis; presynthesized receptors in cell extracts were not affected. Upon withdrawal of geldanamycin, glucocorticoid binding ability was regained; this was partially independent of de novo protein synthesis. Geldanamycin caused decreased levels of immunoreactive glucocorticoid receptors in wild-type cells with enhanced degradation occurring through the ubiquitin-proteasome pathway. Analysis of receptors from treated cells revealed a heteromeric structure of normal size in which the receptor polypeptide is complexed with normal amounts of hsp90 and the immunophilin p59. These data support the view that hsp90 actively participates in steroid-induced signal transduction, and they suggest that geldanamycin affects receptor action without disrupting hsp90-containing heterocomplexes per se. Nevertheless, complexes synthesized and assembled in vitro in the presence of geldanamycin differ from receptors of cellular origin. PMID- 9228041 TI - CYP26, a novel mammalian cytochrome P450, is induced by retinoic acid and defines a new family. AB - A novel member of the cytochrome P450 superfamily, CYP26, which represents a new family of cytochrome P450 enzymes, has been cloned. CYP26 mRNA is up-regulated during the retinoic acid (RA)-induced neural differentiation of mouse embryonic stem cells in vitro and is transiently expressed by embryonic stem cells undergoing predominantly non-neural differentiation. CYP26 transcript is detectable as early as embryonic day 8.5 in mouse embryos, suggesting a function for the gene in early development. CYP26 is expressed in mouse and human liver, as expected for a cytochrome P450, and is also expressed in regions of the brain and the placenta. Acute administration of 100 mg/kg all-trans-RA increases steady state levels of transcript in the adult liver, but not in the brain. CYP26 is highly homologous to a Zebrafish gene, CYPRA1, which has been proposed to participate in the degradation of RA, but is minimally homologous to other mammalian cytochrome P450 proteins. Thus, we report the cloning of a member of a novel cytochrome P450 family that is expressed in mammalian embryos and in brain and is induced by RA in the liver. PMID- 9228042 TI - Characterization of epiphycan, a small proteoglycan with a leucine-rich repeat core protein. AB - The epiphysis of developing bones is a cartilaginous structure that is eventually replaced by bone during skeletal maturation. We have separated a dermatan sulfate proteoglycan, epiphycan, from decorin and biglycan by using dissociative extraction of bovine fetal epiphyseal cartilage, followed by sequential ion exchange, gel permeation, hydrophobic, and Zn2+ chelate chromatographic steps. Epiphycan is a member of the small leucine-rich proteoglycan family, contains seven leucine-rich repeats (LRRs), is related to osteoglycin (osteoinductive factor) (Bentz, H., Nathan, R. M., Rosen, D. M., Armstrong, R. M., Thompson, A. Y., Segarini, P. R., Mathews, M. C., Dasch, J., Piez, K. A., and Seyedin, S. M. (1989) J. Biol. Chem. 264, 20805-20810), and appears to be the bovine equivalent of the chick proteoglycan PG-Lb (Shinomura, T., and Kimata, K. (1992) J. Biol. Chem. 267, 1265-1270). The intact proteoglycan had a median size of approximately 133 kDa. The core protein was 46 kDa by electrophoretic analysis, had a calculated size of 34,271 Da, and had two approximately equimolar N termini (APTLES ... and ETYDAT ... ) separated by 11 amino acids. There were at least three O-linked oligosaccharides in the N-terminal region of the protein, based on blank cycles in Edman degradation and corresponding serine or threonine residues in the translated cDNA sequence. The glycosaminoglycans ranged in size from 23 to 34 kDa were more heterogeneous than those in other dermatan sulfate small leucine rich proteoglycans and were found in the acidic N-terminal region of the protein core, N-terminal to the LRRs. A four-cysteine cluster was present at the N terminus of the LRRs, and a disulfide-bonded cysteine pair was present at the C terminus of the protein core. The seventh LRR and an N-linked oligosaccharide were between the two C-terminal cysteines. An additional potential N glycosylation site near the C terminus did not appear to be substituted at a significant level. PMID- 9228043 TI - Bimodal regulation of ceramidase by interleukin-1beta. Implications for the regulation of cytochrome p450 2C11. AB - Interleukin 1beta (IL-1beta) induces the hydrolysis of sphingomyelin (SM) to ceramide (Cer) in primary cultures of rat hepatocytes, and Cer has been proposed to play a role in the down-regulation of cytochrome P450 2C11 (CYP2C11) and induction of alpha1-acid glycoprotein (AGP) by this cytokine (Chen, J., Nikolova Karakashian, M., Merrill, A. H. & Morgan, E. T. (1995) J. Biol. Chem. 270, 25233 25238). Nonetheless, some of the features of the down-regulation of CYP2C11 do not fit a simple model of Cer as a second messenger as follows: N acetylsphinganine (C2-DHCer) is as potent as N-acetylsphingosine (C2-Cer) in suppression of CYP2C11; the IL-1beta concentration dependence for SM turnover is different from that for the increase in Cer; and the increase in Cer mass is not equivalent to the amount of SM hydrolyzed nor the time course of SM hydrolysis. In this article, we report that these discrepancies are due to activation of ceramidase by the low concentrations of IL-1beta ( approximately 2.5 ng/ml) that maximally down-regulate CYP2C11 expression, whereas higher IL-1beta concentrations (that induce AGP) do not activate ceramidase and allow Cer accumulation. This bimodal concentration dependence is demonstrated both by in vitro ceramidase assays and in intact hepatocytes using a fluorescence Cer analog, 6-((N-(7-nitrobenz-2-oxa-1, 3-diazol-4-yl)amino)-Cer (NBD-Cer), and following release of the NBD-fatty acid. IL-1beta increases both acid and neutral ceramidase activities, which appear to be regulated by tyrosine phosphorylation because pretreatment of hepatocytes with sodium vanadate increases (and 25 microM genistein reduces) the basal and IL-1beta-stimulated ceramidase activities. Since these findings suggested that sphingosine (and, possibly, subsequent metabolites) is the primary mediator of the down-regulation of CYP2C11 by IL-1beta, the effects of exogenous sphingosine and C2-Cer on expression of this gene were compared. Sphingosine was more potent than C2-Cer in down-regulation of CYP2C11 when added alone or with fumonisin B1 to block acylation of the exogenous sphingosine. Furthermore, the suppression of CYP2C11 by C2-Cer (and C2-DHCer) is probably mediated by free sphingoid bases, rather than the short chain Cer directly, because both are hydrolyzed by hepatocytes and increase cellular levels of sphingosine and sphinganine. From these observations we conclude that sphingosine, possibly via sphingosine 1-phosphate, is a mediator of the regulation of CYP2C11 by IL-1beta in rat hepatocytes and that ceramidase activation provides a "switch" that determines which sphingolipids are elevated by this cytokine to produce multiple intracellular responses. PMID- 9228044 TI - Mitochondrial protein import. Tom40 plays a major role in targeting and translocation of preproteins by forming a specific binding site for the presequence. AB - During preprotein transport across the mitochondrial outer membrane, the N terminal presequence initially binds to a surface-exposed site, termed cis site, of the protein translocation complex of this membrane (the TOM complex). The presequence then moves into the translocation pore and becomes exposed at the intermembrane space side. Membrane passage is driven by specific interaction of the presequence with the trans site. We have used chemical cross-linking to identify components in the vicinity of the translocating presequence. Preproteins bound to the surface-exposed cis site can be cross-linked via their N-terminal presequence to Tom20 and Tom22, demonstrating their direct association with this part of the preprotein. In addition, the presequence establishes an early contact to Tom40, a membrane-embedded protein of the TOM complex. Upon further entry of the preprotein into the translocation pore, the presequence loses its contact with Tom20/Tom22, but remains in firm association with Tom40. Our study suggests that Tom40 plays an important function in guiding the presequence of a preprotein across the mitochondrial outer membrane. We propose that Tom40 forms a major part of the trans presequence binding site. PMID- 9228045 TI - Serine phosphorylation within a concise amino-terminal domain in nuclear respiratory factor 1 enhances DNA binding. AB - Nuclear respiratory factor 1 (NRF-1) is a transcriptional activator that acts on a diverse set of nuclear genes required for mitochondrial respiratory function in mammalian cells. These genes encode respiratory proteins as well as components of the mitochondrial transcription, replication, and heme biosynthetic machinery. Here, we establish that NRF-1 is a phosphoprotein in vivo. Phosphorylation occurs on serine residues within a concise NH2-terminal domain with the major sites of phosphate incorporation at serines 39, 44, 46, 47, and 52. The in vivo phosphorylation pattern can be approximated in vitro by phosphorylating recombinant NRF-1 with purified casein kinase II. Phosphate incorporation at the sites utilized in vivo results in a marked stimulation of DNA binding activity which is not observed in mutated proteins lacking these sites. Pairwise expression of the wild-type protein with each of a series of truncated derivatives in transfected cells results in the formation of a dimer between wild type and mutant forms demonstrating that a homodimer is the active binding species. Although NRF-1 can dimerize in the absence of DNA, phosphorylation does not enhance the formation of these dimers. These findings suggest that phosphorylation results in an intrinsic change in the NRF-1 dimer enhancing its ability to bind DNA. PMID- 9228046 TI - Antisera directed against anti-histone H4 antibodies recognize linker histones. Novel immunological probes to detect histone interactions. AB - We introduce a novel immunological approach to detect structural interactions between chromosomal proteins. Antigenically pure core histone H4 was prepared from chicken erythrocytes and used to produce anti-histone H4 antisera. IgG fractions were isolated from purified anti-H4 antibodies and used as antigens to produce "second generation" antisera. Epitopes cross-reacting with the second generation antisera were then identified within chromosomal proteins. These epitopes were presumed to mimic the complementary molecular surface of the original anti-H4 antibodies, and thus proteins containing these epitopes were putatively identified as specific ligands of H4 in chromatin. Surprisingly, we found this immunoreactivity was predominantly directed against H1 compared with H5 from chicken erythrocytes. Further, the immunoreactive epitopes were located within the C-terminal tail domain of the linker histones. These results suggest similar complementary interactions occur between H4 and the C-terminal tail domain of H1s in native chromatin. This could occur either within a single nucleosome as suggested by a previous report (Baneres, J.-L., Essalouh, L., Jariel-Encontre, I., Mesnier, D., Garrod, S., and Parello, J. (1994) J. Mol. Biol., 243, 48-59) or between neighboring nucleosomes within the condensed chromatin fiber. The implications of these results with regard to the structure of the chromatin fiber and the future utility of this technique are discussed. PMID- 9228047 TI - Activation of polyamine catabolism profoundly alters tissue polyamine pools and affects hair growth and female fertility in transgenic mice overexpressing spermidine/spermine N1-acetyltransferase. AB - We have generated a transgenic mouse line that overexpresses the rate-controlling enzyme of polyamine catabolism, spermidine/spermine N1-acetyltransferase. Tissues of these mice showed markedly distorted polyamine pools, which in most cases were characterized by the appearance of N1-acetylspermidine, not normally found in mouse tissues, a massive accumulation of putrescine, and decreases in spermidine and/or spermine pools. The most striking phenotypic change was permanent hair loss at the age of 3 to 4 weeks which was typified histologically by the appearance of extensive follicular cysts in the dermis. The effect seemed attributable to putrescine interference with hair development, possibly with differentiation/proliferation of epidermal cells located in hair follicles. Female members of the transgenic line were found to be infertile apparently due to ovarian hypofunction and hypoplastic uteri. The findings demonstrate the utility of spermidine/spermine N1-acetyltransferase overexpression as an effective means for genetically modulating total tissue polyamine pools in transgenic animals and examining the developmental and oncogenic consequences. PMID- 9228048 TI - Analysis of the dihydropyridine receptor site of L-type calcium channels by alanine-scanning mutagenesis. AB - The dihydropyridine Ca2+ antagonist drugs used in the therapy of cardiovacular disorders inhibit L-type Ca2+ channels by binding to a single high affinity site. Photoaffinity labeling and analysis of mutant Ca2+ channels implicate the IIIS6 and IVS6 segments in high affinity binding. The amino acid residues that are required for high affinity binding of dihydropyridine Ca2+ channel antagonists were probed by alanine-scanning mutagenesis of the alpha1C subunit, transient expression in mammalian cells, and analysis by measurements of ligand binding and block of Ba2+ currents through expressed Ca2+ channels. Eleven amino acid residues in transmembrane segments IIIS6 and IVS6 were identified whose mutation reduced the affinity for the Ca2+ antagonist PN200-110 by 2-25-fold. Both amino acid residues conserved among Ca2+ channels and those specific to L-type Ca2+ channels were found to be required for high affinity dihydropyridine binding. In addition, mutation F1462A increased the affinity for the dihydropyridine Ca2+ antagonist PN200-110 by 416-fold with no effect on the affinity for the Ca2+ agonist Bay K8644. The residues in transmembrane segments IIIS6 and IVS6 that are required for high affinity binding are primarily aligned on single faces of these two alpha helices, supporting a "domain interface model" of dihydropyridine binding and action in which the IIIS6 and IVS6 interact to form a high affinity dihydropyridine receptor site on L-type Ca2+ channels. PMID- 9228049 TI - Molecular determinants of high affinity phenylalkylamine block of L-type calcium channels in transmembrane segment IIIS6 and the pore region of the alpha1 subunit. AB - Recent studies of the phenylalkylamine binding site in the alpha1C subunit of L type Ca2+ channels have revealed three amino acid residues in transmembrane segment IVS6 that are critical for high affinity block and are unique to L-type channels. We have extended this analysis of the phenylalkylamine binding site to amino acid residues in transmembrane segment IIIS6 and the pore region. Twenty two consecutive amino acid residues in segment IIIS6 were mutated to alanine and the conserved Glu residues in the pore region of each homologous domain were mutated to Gln. Mutant channels were expressed in tsA-201 cells along with the beta1b and alpha2delta auxiliary subunits. Assay for block of Ba2+ current by (-) D888 at -60 mV revealed that mutation of five amino acid residues in segment IIIS6 and the pore region that are conserved between L-type and non-L-type channels (Tyr1152, Phe1164, Val1165, Glu1118, and Glu1419) and one L-type specific amino acid (Ile1153) decreased affinity for (-)-D888 from 10-20-fold. Combination of the four mutations in segment IIIS6 increased the IC50 for block by (-)-D888 to approximately 9 microM, similar to the affinity of non-L-type Ca2+ channels for this drug. These results indicate that there are important determinants of phenylalkylamine binding in both the S6 segments and the pore regions of domains III and IV, some of which are conserved across the different classes of voltage-gated Ca2+ channels. A model of the phenylalkylamine receptor site at the interface between domains III and IV of the alpha1 subunit is presented. PMID- 9228050 TI - Mutation of a protease-sensitive region in firefly luciferase alters light emission properties. AB - Luciferase (EC 1.13.12.7) from the North American firefly, Photinus pyralis, is widely used as a reporter enzyme in cell biology. One of its distinctive properties is a pronounced susceptibility to proteolytic degradation that causes luciferase to have a very short intracellular half-life. To define the structural basis for this behavior and possibly facilitate the design of more stable forms of luciferase, limited proteolysis studies were undertaken using trypsin and chymotrypsin to identify regions of the protein whose accessible and flexible character rendered them especially sensitive to cleavage. Regions of amino acid sequence 206-220 and 329-341 were found to be sensitive, and because the region around 206-220 had high homology with other luciferases, CoA ligases, and peptidyl synthetases, this region was selected for mutagenesis experiments intended to determine which of its amino acids were essential for activity. Surprisingly, many highly conserved residues including Ser198, Ser201, Thr202, and Gly203 could be mutated with little effect on the luminescent activity of P. pyralis luciferase. One mutation, however, S198T, caused several alterations in enzymatic properties including shifting the pH optimum from 8.1 to 8.7, lowering the Km for Mg-ATP by a factor of 4 and increasing the half-time for light emission decay by a factor of up to 150. While the S198T luciferase was less active than wild type, activity could be restored by the introduction of the additional L194F and N197Y mutations. In addition to indicating the involvement of this region in ATP binding, these results provide a new form of the enzyme that affords a more versatile reporter system. PMID- 9228051 TI - Studies of the nature of the binding by albumin of platelet-activating factor released from cells. AB - This report confirms that human umbilical vein endothelial cells activated by A23187 produce platelet-activating factor (PAF) (22.4 +/- 9.9 ng/10(6) cells/h; mean +/- S.E.). A proportion of the PAF produced (56%) was released by cells into the medium. The PAF released, however, was not detected without prior organic extraction, and the method of organic extraction was critical for detection. Extraction with 80% ethanol was not successful, but a modified methanol/chloroform extraction method was. These observations may explain some of the conflicting reports in the literature on release of PAF by activated endothelial cells. The requirements for organic extraction may reflect the nature of cell-released PAF's binding by albumin; it was observed that PAF added to identical media could be detected in a bioassay without the requirement for extraction. Such PAF was also readily degraded by PAF-acetylhydrolase added to media, while PAF released from cells was resistant to such degradation, suggesting that it was released in a "protected" configuration. Stimulation of cells was performed in media with albumin as the only extracellular macromolecule. Limited proteolytic digestion of the albumin with trypsin and pepsin showed that PAF released by cells was located exclusively between amino acids 240 and 386 (domain II), while no synthetic PAF added to media was located on this region. These results are identical to those described for the release of PAF by the early embryo. Albumin exposed to embryos had a higher thiol concentration (0.77 +/- 0.04 micromol of thiol/micromol of albumin; mean +/- S.E.) than control media to which an equivalent amount of synthetic PAF was added (0.59 +/- 0.02 micromol of thiol/micromol of albumin) (measured with Ellman's reagent). Furthermore, albumin from conditioned media was more susceptible to reduction by 10 mM dithiothreitol than control albumin, as assessed by its mobility on PAGE. The protected configuration of released PAF was caused by cell dependent conformational changes to albumin involving cysteine-cysteine disulfide bonds. Partial reduction with dithiothreitol of albumin exposed to cells resulted in released PAF being able to be detected directly in a bioassay without the requirement for prior organic extraction. PMID- 9228052 TI - The organization, promoter analysis, and expression of the human PPARgamma gene. AB - PPARgamma is a member of the PPAR subfamily of nuclear receptors. In this work, the structure of the human PPARgamma cDNA and gene was determined, and its promoters and tissue-specific expression were functionally characterized. Similar to the mouse, two PPAR isoforms, PPARgamma1 and PPARgamma2, were detected in man. The relative expression of human PPARgamma was studied by a newly developed and sensitive reverse transcriptase-competitive polymerase chain reaction method, which allowed us to distinguish between PPARgamma1 and gamma2 mRNA. In all tissues analyzed, PPARgamma2 was much less abundant than PPARgamma1. Adipose tissue and large intestine have the highest levels of PPARgamma mRNA; kidney, liver, and small intestine have intermediate levels; whereas PPARgamma is barely detectable in muscle. This high level expression of PPARgamma in colon warrants further study in view of the well established role of fatty acid and arachidonic acid derivatives in colonic disease. Similarly as mouse PPARgammas, the human PPARgammas are activated by thiazolidinediones and prostaglandin J and bind with high affinity to a PPRE. The human PPARgamma gene has nine exons and extends over more than 100 kilobases of genomic DNA. Alternate transcription start sites and alternate splicing generate the PPARgamma1 and PPARgamma2 mRNAs, which differ at their 5'-ends. PPARgamma1 is encoded by eight exons, and PPARgamma2 is encoded by seven exons. The 5'-untranslated sequence of PPARgamma1 is comprised of exons A1 and A2, whereas that of PPARgamma2 plus the additional PPARgamma2-specific N terminal amino acids are encoded by exon B, located between exons A2 and A1. The remaining six exons, termed 1 to 6, are common to the PPARgamma1 and gamma2. Knowledge of the gene structure will allow screening for PPARgamma mutations in humans with metabolic disorders, whereas knowledge of its expression pattern and factors regulating its expression could be of major importance in understanding its biology. PMID- 9228053 TI - Ras2 and Ras1 protein phosphorylation in Saccharomyces cerevisiae. AB - This work describes the phosphorylation of Saccharomyces cerevisiae Ras proteins and explores the physiological role of the phosphorylation of Ras2 protein. Proteins expressed from activated alleles of RAS were less stable and less phosphorylated than proteins from cells expressing wild-type alleles of RAS. This difference in phosphorylation level did not result from increased signaling through the Ras-cAMP pathway or reflect the primarily GTP-bound nature of activated forms of Ras protein per se. In addition, phosphorylation of Ras protein was not dependent on proper localization of the Ras2 protein to the plasma membrane nor on the interaction of Ras2p with its exchange factor, Cdc25p. The preferred phosphorylation site on Ras2 protein was identified as serine 214. This site, when mutated to alanine, led to promiscuous phosphorylation of Ras2 protein on nearby serine residues. A decrease in phosphorylation may lead to a decrease in signaling through the Ras-cAMP pathway. PMID- 9228054 TI - The Ca2+-dependent binding of calmodulin to an N-terminal motif of the heterotrimeric G protein beta subunit. AB - Ca2+ ion concentration changes are critical events in signal transduction. The Ca2+-dependent interactions of calmodulin (CaM) with its target proteins play an essential role in a variety of cellular functions. In this study, we investigated the interactions of G protein betagamma subunits with CaM. We found that CaM binds to known betagamma subunits and these interactions are Ca2+-dependent. The CaM-binding domain in Gbetagamma subunits is identified as Gbeta residues 40-63. Peptides derived from the Gbeta protein not only produce a Ca2+-dependent gel mobility shifting of CaM but also inhibit the CaM-mediated activation of CaM kinase II. Specific amino acid residues critical for the binding of Gbetagamma to CaM were also identified. We then investigated the potential function of these interactions and showed that binding of CaM to Gbetagamma inhibits the pertussis toxin-catalyzed ADP-ribosylation of Galphao subunits, presumably by inhibiting heterotrimer formation. Furthermore, we demonstrated that interaction with CaM has little effect on the activation of phospholipase C-beta2 by Gbetagamma subunits, supporting the notion that different domains of Gbetagamma are responsible for the interactions of different effectors. These findings shed light on the molecular basis for the interactions of Gbetagamma with Ca2+-CaM and point to the potential physiological significance of these interactions in cellular functions. PMID- 9228055 TI - Cellular respiration during hypoxia. Role of cytochrome oxidase as the oxygen sensor in hepatocytes. AB - We previously reported that hepatocytes exhibit a reversible suppression of respiration during prolonged hypoxia (PO2 = 20 torr for 3-5 h). Also, isolated bovine heart cytochrome c oxidase undergoes a reversible decrease in apparent Vmax when incubated under similar conditions. This study sought to link the hypoxia-induced changes in cytochrome oxidase to the inhibition of respiration seen in intact cells. Hepatocytes incubated at PO2 = 20 torr exhibited decreases in respiration and increases in [NAD(P)H] after 2-3 h that were reversed upon reoxygenation (PO2 = 100 torr). Respiration during hypoxia was also inhibited when N,N,N',N'-tetramethyl-p-phenylenediamine (0.5 mM) and ascorbate (5 mM) were used to reduce cytochrome c, suggesting that cytochrome oxidase was partially inhibited. Similarly, liver submitochondrial particles revealed a 44% decrease in the apparent Vmax of cytochrome oxidase after hypoxic incubation. In hepatocytes loaded with tetramethylrhodamine ethyl ester (10 nM) to quantify mitochondrial membrane potential, acute hypoxia (<30 min) produced no change in fluorescence, consistent with the absence of an acute change in respiration. However, fluorescence increased during acute reoxygenation after prolonged hypoxia, suggesting an increase in potential. The control exhibited by NADH over mitochondrial respiration was not altered during hypoxia. Thus, changes in the Vmax of cytochrome oxidase during prolonged hypoxia correlate with the changes in respiration and mitochondrial potential. This suggests that the oxidase functions as an oxygen sensor in the intact hepatocyte. PMID- 9228056 TI - The potential roles of the conserved amino acids in human liver mitochondrial aldehyde dehydrogenase. AB - The sequence alignment of all known aldehyde dehydrogenases showed that only 23 residues were completely conserved (Hempel, J., Nicholas, H., and Lindahl, R. (1993) Protein Sci. 2, 1890-1900). Of these 14 were glycines and prolines. Site directed mutagenesis showed that Cys302 was the essential nucleophile and that Glu268 was the general base necessary to activate Cys302 for both the dehydrogenase and esterase reaction. Here we report the mutational analysis of other conserved residues possessing reactive side chains Arg84, Lys192, Thr384, Glu399, and Ser471, along with partially conserved Glu398 and Lys489, to determine their involvement in the catalytic process and correlate these finding with the known structure of mitochondrial ALDH (Steinmetz, C. G., Xie, P.-G., Weiner, H., and Hurley, T. D. (1997) Structure 5, 701-711). No residue was found to be absolutely essential, but all the mutations caused a decrease in the specific activity of the enzyme. None of the mutations affected the Km for aldehyde significantly, although k3, the rate constant calculated for aldehyde binding was decreased. The Km and dissociation constant (Kia) for NAD+ increased significantly for K192Q and S471A compared with the native enzyme. Mutations of only Lys192 and Glu399, both NAD+-ribose binding residues, led to a change in the rate-limiting step such that hydride transfer became rate-limiting, not deacylation. Esterase activity of all mutants decreased even though mutations affected different catalytic steps in the dehydrogenase reaction. PMID- 9228057 TI - Involvement of glutamate 399 and lysine 192 in the mechanism of human liver mitochondrial aldehyde dehydrogenase. AB - Mutation to the conserved Glu399 or Lys192 caused the rate-limiting step of human liver mitochondrial aldehyde dehydrogenase (ALDH2) to change from deacylation to hydride transfer (Sheikh, S., Ni, L., Hurley, T. D., and Weiner, H. (1997) J. Biol. Chem. 272, 18817-18822). Here we further investigated the role of these two NAD+-ribose-binding residues. The E399Q/K/H/D and K192Q mutants had lower dehydrogenase activity when compared with the native enzyme. No pre-steady state burst of NADH formation was found with the E399Q/K and K192Q enzymes when propionaldehyde was used as the substrate; furthermore, each mutant oxidized chloroacetaldehyde slower than propionaldehyde, and a primary isotope effect was observed for each mutant when [2H]acetaldehyde was used as a substrate. However, no isotope effect was observed for each mutant when alpha-[2H]benzaldehyde was the substrate. A pre-steady state burst of NADH formation was observed for the E399Q/K and K192Q mutants with benzaldehyde, and p-nitrobenzaldehyde was oxidized faster than benzaldehyde. Hence, when aromatic aldehydes were used as substrates, the rate-limiting step remained deacylation for all these mutants. The rate limiting step remained deacylation for the E399H/D mutants when either aliphatic or aromatic aldehydes were used as substrates. The K192Q mutant displayed a change in substrate specificity, with aromatic aldehydes becoming better substrates than aliphatic aldehydes. PMID- 9228058 TI - Characterization of Fas (Apo-1, CD95)-Fas ligand interaction. AB - The death-inducing receptor Fas is activated when cross-linked by the type II membrane protein Fas ligand (FasL). When human soluble FasL (sFasL, containing the extracellular portion) was expressed in human embryo kidney 293 cells, the three N-linked glycans of each FasL monomer were found to be essential for efficient secretion. Based on the structure of the closely related lymphotoxin alpha-tumor necrosis factor receptor I complex, a molecular model of the FasL homotrimer bound to three Fas molecules was generated using knowledge-based protein modeling methods. Point mutations of amino acid residues predicted to affect the receptor-ligand interaction were introduced at three sites. The F275L mutant, mimicking the loss of function murine gld mutation, exhibited a high propensity for aggregation and was unable to bind to Fas. Mutants P206R, P206D, and P206F displayed reduced cytotoxicity toward Fas-positive cells with a concomitant decrease in the binding affinity for the recombinant Fas immunoglobulin Fc fusion proteins. Although the cytotoxic activity of mutant Y218D was unaltered, mutant Y218R was inactive, correlating with the prediction that Tyr-218 of FasL interacts with a cluster of three basic amino acid side chains of Fas. Interestingly, mutant Y218F could induce apoptosis in murine, but not human cells. PMID- 9228060 TI - Molecular cloning of a novel polypeptide, DP5, induced during programmed neuronal death. AB - To study the molecular mechanisms underlying neuronal programmed cell death (PCD), we performed differential display screening for genes, the expression of which was induced during PCD in the sympathetic neuron culture model deprived of NGF. We cloned a gene encoding a novel polypeptide (DP5) which consisted of 92 amino acids. DP5 polypeptide had no homology with any other known protein and contained no motif that would indicate its putative biochemical functions. DP5 mRNA levels peaked at 15 h after nerve growth factor withdrawal, concurrent with the time at which neurons were committed to die. The induction of DP5 gene expression was blocked when cell death was rescued by treatment with cycloheximide, KCl, or the cyclic AMP analogue CPTcAMP. Overexpression of the full-length DP5 in cultured sympathetic neurons was in itself sufficient to induce apoptosis. These results suggest that DP5 plays a role in programmed neuronal death. PMID- 9228059 TI - Rac binding to p67(phox). Structural basis for interactions of the Rac1 effector region and insert region with components of the respiratory burst oxidase. AB - Activation of the respiratory burst oxidase involves the assembly of the membrane associated flavocytochrome b558 with the cytosolic components p47(phox), p67(phox), and the small GTPase Rac. Herein, the interaction between Rac and p67(phox) is explored using functional and physical methods. Mutually facilitated binding (EC50) of Rac1 and p67(phox) within the NADPH oxidase complex was demonstrated using steady state kinetic methods measuring NADPH-dependent superoxide generation. Direct binding of Rac1 and Rac2 to p67(phox) was shown using a fluorescent analog of GTP (methylanthraniloyl guanosine-5'-[beta,gamma imido]triphosphate) bound to Rac as a reporter group. An increase in the methylanthraniloyl fluorescence was seen with added p67(phox) but not p47(phox), and the emission maximum shifted from 445 to 440 nm. Rac1 and Rac2 bound to p67(phox) with a 1:1 stoichiometry and with Kd values of 120 and 60 nM, respectively. Mutational studies (Freeman, J., Kreck, M., Uhlinger, D. J., and Lambeth, J. D. (1994) Biochemistry 33, 13431-13435; Freeman, J. L., Abo, A., and Lambeth, J. D. (1996) J. Biol. Chem. 271, 19794-19801) previously identified two regions in Rac1 that are important for activity: the "effector region" (residues 26-45) and the "insert region" (residues 124-135). Proteins mutated in the effector region (Rac1(N26H), Rac1(I33N), and Rac1(D38N)) showed a marked increase in both the Kd and the EC50, indicating that mutations in this region affect activity by inhibiting Rac binding to p67(phox). Insert region mutations (Rac1(K132E) and L134R), while showing markedly elevated EC50 values, bound with normal affinity to p67(phox). The structure of Rac1 determined by x-ray crystallography reveals that the effector region and the insert region are located in defined sectors on the surface of Rac1. A model is discussed in which the Rac1 effector region binds to p67(phox), the C terminus binds to the membrane, and the insert region interacts with a different protein component, possibly cytochrome b558. PMID- 9228061 TI - Three discrete regions of mammalian adenylyl cyclase form a site for Gsalpha activation. AB - The interaction between the alpha subunit of G protein Gs (Gsalpha) and the two cytoplasmic domains of adenylyl cyclase (C1 and C2) is a key step in the stimulation of cAMP synthesis by hormones. Mutational analysis reveals that three discrete regions in the primary sequence of adenylyl cyclase affect the EC50 values for Gsalpha activation and thus are the affinity determinants of Gsalpha. Based on the three-dimensional structure of C2.forskolin dimer, these three regions (C2 alpha2, C2 alpha3/beta4, and C1 beta1) are close together and form a negatively charged and hydrophobic groove the width of an alpha helix that can accommodate the positively charged adenylyl cyclase binding region of Gsalpha. Two mutations in the C2 alpha3/beta4 region decrease the Vmax values of Gsalpha activation without an increase in the EC50 values. Since these three regions are distal to the catalytic site, the likely mechanism for Gsalpha activation is to modulate the structure of the active site by controlling the orientation of the C2 alpha2 and alpha3/beta4 structures. PMID- 9228062 TI - Engineering of porcine pepsin. Alteration of S1 substrate specificity of pepsin to those of fungal aspartic proteinases by site-directed mutagenesis. AB - The S1 substrate specificity of porcine pepsin has been altered to resemble that of fungal aspartic proteinase with preference for a basic amino acid residue in P1 by site directed mutagenesis. On the basis of primary and tertiary structures of aspartic proteinases, the active site-flap mutants of porcine pepsin were constructed, which involved the replacement of Thr-77 by Asp (T77D), the insertion of Ser between Gly-78 and Ser-79 (G78(S)S79), and the double mutation (T77D/G78(S)S79). The specificities of the mutants were determined using p nitrophenylalanine-based substrates containing a Phe or Lys residue at the P1 position. The double mutant cleaved the Lys-Phe(4-NO2) bonds, while wild-type enzyme digested other bonds. In addition, the pH dependence of hydrolysis of Lys containing substrates by the double mutant indicates that the interactions between Asp-77 of the mutant and P1 Lys contribute to the transition state stabilization. The double mutant was also able to activate bovine trypsinogen to trypsin by the selective cleavage of the Lys6-Ile7 bond of trypsinogen. Results of this study suggest that the structure of the active site flap contributes to the S1 substrate specificity for basic amino acid residues in aspartic proteinases. PMID- 9228063 TI - Ectopic expression of phospholamban in fast-twitch skeletal muscle alters sarcoplasmic reticulum Ca2+ transport and muscle relaxation. AB - There are three isoforms of the sarcoplasmic reticulum Ca2+-ATPase; they are known as SERCA1, SERCA2, and SERCA3. Phospholamban is present in tissues that express the SERCA2 isoform and is an inhibitor of the affinity of SERCA2 for calcium. In vitro reconstitution and cell culture expression studies have shown that phospholamban can also regulate SERCA1, the fast-twitch skeletal muscle isoform. To determine whether regulation of SERCA1 by phospholamban can be of physiological relevance, we generated transgenic mice that ectopically express phospholamban in fast-twitch skeletal muscle, a tissue normally devoid of phospholamban. Ectopic expression of phospholamban was associated with a decrease in the affinity of SERCA1 for calcium. Assessment of isometric twitch contractions of intact fast-twitch skeletal muscles revealed depressed rates of relaxation in transgenic mice compared with wild-type cohorts. Furthermore, the prolongation of muscle relaxation appeared to correlate with the levels of phospholamban expressed in two transgenic mouse lines. These findings indicate that ectopic expression of phospholamban in fast-twitch skeletal muscle is associated with inhibition of SERCA1 activity and decreased relaxation rates of this muscle. PMID- 9228064 TI - Identification of CDK4 sequences involved in cyclin D1 and p16 binding. AB - Activation of CDK4 is regulated, in part, by its association with a D-type cyclin. Conversely, CDK4 activity is inhibited when it is bound to the cyclin dependent kinase inhibitor, p16(INK4A). To investigate the molecular basis of the interactions between CDK4 and cyclin D1 or p16(INK4A) we performed site-directed mutagenesis of CDK4. The interaction was examined using in vitro translated wild type and mutant CDK4 proteins and bacterially expressed cyclin D1 and p16 fusion proteins. As mutational analysis of CDC2 suggests that its cyclin binding domain is primarily located near its amino terminus, the majority of the mutations constructed in CDK4 were located near its amino terminus. In addition, CDK4 residues homologous to CDC2 sites involved in Suc1 binding were also mutated. Our analysis indicates that cyclin D1 and p16 binding sites are overlapping and located primarily near the amino terminus. All CDK4 mutations that resulted in decreased p16 binding capability also diminished cyclin D1 binding. In contrast, amino-terminal sequences were identified, including the PSTAIRE region, that are important for cyclin D1 binding but are not involved in p16 binding. PMID- 9228065 TI - Novel insights into the chemical mechanism of ATP synthase. Evidence that in the transition state the gamma-phosphate of ATP is near the conserved alanine within the P-loop of the beta-subunit. AB - The chemical mechanism by which the F1 moiety of ATP synthase hydrolyzes and synthesizes ATP remains unknown. For this reason, we have carried out studies with orthovanadate (Vi), a phosphate analog which has the potential of "locking" an ATPase, in its transition state by forming a MgADP.Vi complex, and also the potential, in a photochemical reaction resulting in peptide bond cleavage, of identifying an amino acid very near the gamma-phosphate of ATP. Upon incubating purified rat liver F1 with MgADP and Vi for 2 h to promote formation of a MgADP.Vi-F1 complex, the ATPase activity of the enzyme was markedly inhibited in a reversible manner. When the resultant complex was formed in the presence of ultraviolet light inhibition could not be reversed, and SDS-polyacrylamide gel electrophoresis revealed, in addition to the five known subunit bands characteristic of F1 (i.e. alpha, beta, gamma, delta, and epsilon), two new electrophoretic species of 17 and 34 kDa. Western blot and N-terminal sequencing analyses identified both bands as arising from the beta subunit with the site of peptide bond cleavage occurring at alanine 158, a conserved residue within F1 ATPases and the third residue within the nucleotide binding consensus GX4GK(T/S) (P-loop). Quantification of the amount of ADP bound within the MgADP. Vi-F1 complex revealed about 1.0 mol/mol F1, while quantification of the peptide cleavage products revealed that no more than one beta subunit had been cleaved. Consistent with the cleavage reaction involving oxidation of the methyl group of alanine was the finding that [3H] from NaB[3H]4 incorporates into MgADP.Vi-F1 complex following treatment with ultraviolet light. These novel findings provide information about the transition state involved in the hydrolysis of ATP by a single beta subunit within F1-ATPases and implicate alanine 158 as residing very near the gamma-phosphate of ATP during catalysis. When considered with earlier studies on myosin and adenylate kinase, these studies also implicate a special role for the third residue within the GX4GK(T/S) sequence of many other nucleotide-binding proteins. PMID- 9228066 TI - Synergistic regulation of m2 muscarinic acetylcholine receptor desensitization and sequestration by G protein-coupled receptor kinase-2 and beta-arrestin-1. AB - The m2 muscarinic acetylcholine receptor (m2 mAChR) belongs to the superfamily of G protein-coupled receptors and is regulated by many processes that attenuate signaling following prolonged stimulation by agonist. We used a heterologous expression system to examine the ability of G protein-coupled receptor kinase-2 (GRK2) and beta-arrestin-1 to regulate the phosphorylation state and to promote desensitization and sequestration of the m2 mAChR. Treatment of JEG-3 cells transiently expressing the m2 mAChR with a muscarinic agonist induced an approximately 4- or 8-fold increase in receptor phosphorylation in the absence or presence of cotransfected GRK2, respectively, compared with untreated cells transfected with receptor alone. Using the expression of a cAMP-regulated reporter gene to measure receptor function, we found that transiently transfected m2 mAChRs underwent functional desensitization following exposure to agonist. Transfected GRK2 enhanced agonist-induced functional desensitization in a manner that was synergistically enhanced by cotransfection of beta-arrestin-1, which had no effect on m2 mAChR function when coexpressed in the absence of GRK2. Finally, GRK2 and beta-arrestin-1 synergistically enhanced both the rate and extent of agonist-induced m2 mAChR sequestration. These results are the first to demonstrate that agonist-induced desensitization and sequestration of the m2 mAChR in the intact cell can be enhanced by the presence of GRK2 and beta arrestin-1 and show that these molecules have multiple actions on the m2 mAChR. PMID- 9228067 TI - Function of the type V transforming growth factor beta receptor in transforming growth factor beta-induced growth inhibition of mink lung epithelial cells. AB - The type V transforming growth factor beta (TGF-beta) is a 400-kDa nonproteoglycan membrane protein that co-expresses with the type I, type II, and type III TGF-beta receptors in most cell types. The type V TGF-beta receptor exhibits a Ser/Thr-specific protein kinase activity with distinct substrate specificity (Liu, Q., Huang, S. S., and Huang, J. (1994) J. Biol. Chem. 269, 9221 9226). In mink lung epithelial cells, the type V TGF-beta receptor was found to form heterocomplexes with the type I TGF-beta receptor by immunoprecipitation with antiserum to the type V TGF-beta receptor after 125I-TGF-beta affinity labeling or Trans35S-label metabolic labeling of the cells. The kinase activity of the type V TGF-beta receptor was stimulated after treatment of mink lung epithelial cells with TGF-beta. TGF-beta stimulation resulted in the growth inhibition of wild-type mink lung epithelial cells and to a lesser extent of the type I and type II TGF-beta receptor-defective mutants, although higher concentrations of TGF-beta were required for the growth inhibition of these mutants. TGF-beta was unable to induce growth inhibition in human colorectal carcinoma cells lacking the type V TGF-beta receptor but expressing the type I and type II TGF-beta receptors. These results suggest that the type V TGF-beta receptor can mediate the TGF-beta-induced growth inhibitory response in the absence of the type I or type II TGF-beta receptor. These results also support the hypothesis that loss of the type V TGF-beta receptor may contribute to the malignancy of certain carcinoma cells. PMID- 9228068 TI - Functional analysis of in vivo and in organello footprinting of HeLa cell mitochondrial DNA in relationship to ATP and ethidium bromide effects on transcription. AB - In vivo and in organello footprinting techniques based on methylation interference have been utilized to investigate protein-DNA interactions in the transcription initiation and rDNA transcription termination regions of human mitochondrial DNA (mtDNA) in functionally active mitochondria. In particular, the changes in methylation reactivity of these regions in response to treatment of the organelles with ATP or ethidium bromide, which affects differentially the rates of mitochondrial rRNA and mRNA synthesis, have been analyzed. Two major sites of protein-DNA interactions have been identified in the main control region of mtDNA, both in vivo and in organello, which correspond to the regions of the light-strand promoter and heavy-strand rRNA-specific promoter. The in organello footprinting of the latter showed ATP- and ethidium bromide-dependent modifications that could be correlated with changes in the rate of rRNA but not of mRNA synthesis. By contrast, no ATP effects were observed on the in organello footprinting pattern of the termination region and on in vitro transcription termination, strongly suggesting that ATP control of rRNA synthesis occurs at the initiation level. Several methylation interference sites were found upstream of the whole H-strand transcription unit, pointing to possible protein-DNA interactions related to the activity of this unit. In vivo footprinting of the rDNA transcription termination region of human mtDNA has revealed a very strong protection pattern, indicating a high degree of occupancy of the termination site by mitochondrial transcription termination factor (approximately 80%). PMID- 9228069 TI - Tyrosine phosphorylation of RNA polymerase II carboxyl-terminal domain by the Abl related gene product. AB - The largest subunit of RNA polymerase II contains a C-terminal repeated domain (CTD) that is the site of phosphorylation by serine (threonine) and tyrosine kinases. Phosphorylation of the CTD is correlated with transcription elongation. A number of different kinases have previously been shown to phosphorylate the CTD; among them is a nuclear tyrosine kinase encoded by the c-abl proto-oncogene. The processive and high stoichiometric phosphorylation of RNA polymerase II by c Abl requires the tyrosine kinase, the SH2 domain, and a CTD-interacting domain (CTD-ID) in the Abl protein. The physiological tyrosine phosphorylation of RNA polymerase II by c-Abl in DNA damage response has previously been demonstrated. Basal tyrosine phosphorylation of RNA polymerase II, however, is observed in cells derived from abl-deficient mice, indicating the existence of other CTD tyrosine kinases. In this report, we show that the tyrosine kinase encoded by an Abl-related gene (Arg) also phosphorylates the CTD in vitro and in transfected cells. The SH2 and kinase domain of Arg are 95% identical to that of c-Abl. However, these two proteins share only 29% identity in the large C-terminal region. Interestingly, a CTD-ID is also found in the C-terminal region of Arg. Mapping studies and sequence analysis have led to the identification of the CTD ID that is highly conserved among the divergent C-terminal regions of Abl and Arg. These results indicate that tyrosine phosphorylation of RNA polymerase II CTD could be catalyzed by either c-Abl or Arg kinase. PMID- 9228070 TI - A DNA helicase from Schizosaccharomyces pombe stimulated by single-stranded DNA binding protein at low ATP concentration. AB - A DNA helicase named DNA helicase I was isolated from cell-free extracts of the fission yeast Schizosaccharomyces pombe. Both DNA helicase and single-stranded DNA-dependent ATPase activities copurified with a polypeptide of 95 kDa on an SDS polyacrylamide gel. The helicase possessed a sedimentation coefficient of 6.0 S and a Stokes radius of 44.8 A determined by glycerol gradient centrifugation and gel filtration analysis, respectively. From these data the native molecular mass was calculated to be 110 kDa, indicating that the active enzyme is a monomer. The DNA-unwinding and ATP hydrolysis activities associated with DNA helicase I have been examined. One notable property of the enzyme was its relatively high rate of ATP turnover (35-50 molecules of ATP hydrolyzed/s/enzyme molecule) that may contribute to its inefficient unwinding activity at low concentrations of ATP (<0.2 mM). Addition of an ATP-regenerating system to the reaction mixture restored the DNA-unwinding activity of the enzyme. S. pombe single-stranded DNA binding protein (SpSSB, also called SpRPA) stimulated the DNA helicase activity significantly at low levels of ATP (0.025-0.2 mM) even in the absence of an ATP regenerating system. In contrast, SpRPA had no effect on ATP hydrolysis at any ATP concentration examined. These observations suggest that the stimulation of DNA unwinding by SpRPA is not simply a result of suppression of nonproductive ATP hydrolysis. Rather, the role of SpRPA is to lower the Km for ATP in the unwinding reaction, allowing the helicase to function efficiently at low ATP concentrations. PMID- 9228071 TI - Phosphocitrate inhibits a basic calcium phosphate and calcium pyrophosphate dihydrate crystal-induced mitogen-activated protein kinase cascade signal transduction pathway. AB - Calcium deposition diseases caused by calcium pyrophosphate dihydrate (CPPD) and basic calcium phosphate (BCP) crystals are a significant source of morbidity in the elderly. We have shown previously that both types of crystals can induce mitogenesis, as well as metalloproteinase synthesis and secretion by fibroblasts and chondrocytes. These responses may promote degradation of articular tissues. We have also shown previously that both CPPD and BCP crystals activate expression of the c-fos and c-jun proto-oncogenes. Phosphocitrate (PC) can specifically block mitogenesis and proto-oncogene expression induced by either BCP or CPPD crystals in 3T3 cells and human fibroblasts, suggesting that PC may be an effective therapy for calcium deposition diseases. To understand how PC inhibits BCP and CPPD-mediated cellular effects, we have investigated the mechanism by which BCP and CPPD transduce signals to the nucleus. Here we demonstrate that BCP and CPPD crystals activate a protein kinase signal transduction pathway involving p42 and p44 mitogen-activated protein (MAP) kinases (ERK 2 and ERK 1). BCP and CPPD also cause phosphorylation of a nuclear transcription factor, cyclic AMP response element-binding protein (CREB), on serine 133, a residue essential for CREB's ability to transactivate. Treatment of cells with PC at concentrations of 10(-3) to 10(-5) M blocked both the activation of p42/p44 MAP kinases, and CREB serine 133 phosphorylation, in a dose-dependent fashion. At 10(-3) M, a PC analogue, n-sulfo-2-aminotricarballylate and citrate also modulate this signal transduction pathway. Inhibition by PC is specific for BCP- and CPPD-mediated signaling, since all three compounds had no effect on serum-induced p42/P44 or interleukin-1beta induced p38 MAP kinase activities. Treatment of cells with an inhibitor of MEK1, an upstream activator of MAPKs, significantly inhibited crystal-induced cell proliferation, suggesting that the MAPK pathway is a significant mediator of crystal-induced signals. PMID- 9228072 TI - A raf-independent epidermal growth factor receptor autocrine loop is necessary for Ras transformation of rat intestinal epithelial cells. AB - We recently have shown that activated Ras, but not Raf, causes transformation of intestinal (RIE-1, IEC-6) epithelial cells, whereas both activated Ras and Raf transform NIH 3T3 fibroblasts (Oldham, S. M., Clark, G. J., Gangarosa, L. M., Coffey, R. J., and Der, C. J. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 6924 6928). The observations that conditioned medium from Ras-, but not Raf-, transfected RIE-1 cells, as well as exogenous transforming growth factor alpha (TGFalpha), promoted morphological transformation of parental RIE-1 cells prompted us to identify epidermal growth factor (EGF) receptor (EGFR) ligands produced by Ras-transformed RIE-1 cells responsible for this autocrine effect. Since studies in fibroblasts have shown that v-Src is transforming, we also determined if v-Src could transform RIE-1 cells. H- or K-Ras-transformed cells secreted significant amounts of TGFalpha protein, and mRNA transcripts for TGFalpha, amphiregulin (AR), and heparin-binding EGF-like growth factor (HB-EGF) were induced. Like Ras, v-Src caused morphological and growth transformation of parental RIE-1 cells. However, TGFalpha protein was not secreted by RIE-1 cells stably expressing v-Src or activated Raf, and only minor increases in EGFR ligand mRNA expression were detected in these cells. A selective EGFR tyrosine kinase inhibitor PD153035 attenuated the Ras-, but not Src-, transformed phenotype. Taken together, these observations provide a mechanistic and biochemical basis for the ability of activated Ras, but not activated Raf, to cause transformation of RIE-1 cells. Finally, we suggest that an EGFR-dependent mechanism is necessary for Ras, but not Src, transformation of these intestinal epithelial cells. PMID- 9228073 TI - The high affinity heparin-binding domain and the V region of fibronectin mediate invasion of human oral squamous cell carcinoma cells in vitro. AB - Fibronectin is an extracellular matrix molecule composed of repeating subunits that create functional domains. These domains contain multiple binding sites for heparin and for various cell-surface receptors that modulate cell function. To examine the role that the high affinity heparin-binding region and the alternatively spliced V region of fibronectin play in tumor invasion, we expressed and purified four complementary recombinant fibronectin proteins. These proteins either included or excluded the alternatively spliced V region and contained either a mutated, non-functional high affinity heparin-binding domain (Hep-) or an unmutated heparin-binding domain (Hep+). Cultured oral squamous cell carcinoma cells were assayed for invasion into a Matrigel/collagen matrix supplemented with these four purified recombinant proteins, and for spreading and motility on plastic. Increased invasion was observed in gels supplemented with the V-Hep+ protein when compared with the V-Hep- protein. Inclusion of the V region in the proteins enhanced the invasion and migration associated with both Hep+ and Hep- proteins, whereas cell spreading was enhanced with the Hep+ recombinant proteins. These data demonstrate that both the high affinity heparin binding domain and the V region of fibronectin play important roles in invasion, motility, and spreading of oral squamous cell carcinoma cells. PMID- 9228074 TI - N-terminal domains of human copper-transporting adenosine triphosphatases (the Wilson's and Menkes disease proteins) bind copper selectively in vivo and in vitro with stoichiometry of one copper per metal-binding repeat. AB - N-terminal domains of the Wilson's and Menkes disease proteins (N-WND and N-MNK) were overexpressed in a soluble form in Escherichia coli as fusions with maltose binding protein, purified, and their metal-binding properties were characterized. Both N-MNK and N-WND bind copper specifically as indicated by the results of metal-chelate chromatography, direct copper-binding measurements, and chemical modification of Cys residues in the presence of different heavy metals. When E. coli cells are grown in the presence of copper, N-MNK and N-WND bind copper in vivo with stoichiometry of 5-6 nmol of copper/nmol of protein. Copper released from the copper-N-MNK and copper-N-WND complexes reacts with the Cu(I)-selective chelator bicinchoninic acid in the absence of reducing agents. This suggests that in proteins, it is bound in reduced Cu(I) form, in agreement with the spectroscopic properties of the copper-bound domains. Copper bound to the domains in vivo or in vitro specifically protects the N-MNK and N-WND against labeling with the cysteine-directed probe; this indicates that Cys residues in the repetitive motifs GMTCXXCXXXIE are involved in coordination of copper. Direct involvement of the N-terminal domains in the binding of copper suggests their important role in copper-dependent functions of human copper-transporting adenosine triphosphatases (Wilson's and Menkes disease proteins). PMID- 9228075 TI - Cloning and subcellular localization of hamster and rat isopentenyl diphosphate dimethylallyl diphosphate isomerase. A PTS1 motif targets the enzyme to peroxisomes. AB - To date, isopentenyl diphosphate:dimethylallyl diphosphate isomerase (IPP isomerase; EC 5.3.3.2) is presumed to have a cytosolic localization. However, we have recently shown that in permeabilized cells lacking cytosolic components, mevalonate can be converted to cholesterol, implying that all of the enzymes required for the conversion of mevalonate to farnesyl diphosphate are found in the peroxisome. To provide unequivocal evidence for the subcellular localization of IPP isomerase, in this study, we have cloned the rat and hamster homologues of IPP isomerase and identified the signal that targets this enzyme to peroxisomes. In addition, we also demonstrate that IPP isomerase is regulated at the mRNA level. PMID- 9228076 TI - Monkey growth hormone (GH) receptor gene expression. Evidence for two mechanisms for the generation of the GH binding protein. AB - The growth hormone receptor (GHR) cDNA was cloned from the liver of Rhesus macaque using polymerase chain reaction. As deduced from the nucleotide sequence, the mature GHR is a protein of 620 amino acids which presents 94.1% identity with the human receptor. The monkey GHR (mkGHR) expressed in 293 cells presented the expected specificity for a primate GHR and was able to transduce a transcriptional effect of GH. Human GH was able to activate tyrosine phosphorylation of both the tyrosine kinase JAK2 and the receptor in 293 cells co transfected with mkGHR and JAK2 cDNAs. The GH binding protein (GHBP), the soluble short form of the GHR, was also present in monkey serum. Expression of the GHR cDNA in eucaryotic cells indicated that the GHBP can be produced by proteolytic cleavage of the membrane receptor. Northern blot analysis of GHR gene expression in different tissues allowed us to identify three different transcripts of 5.0 and 2.8 kilobase pairs and a smaller one of 1.7 kilobase pairs which could encode a GHBP. Rapid amplification of cDNA extremities (3'-RACE-polymerase chain reaction) was used to identify a cDNA encoding a protein in which the transmembrane and cytoplasmic domains of the receptor are substituted by a short sequence of 9 amino acids. This transcript was present in various tissues and could encode a GHBP as well, suggesting for the first time that two different mechanisms can coexist for the generation of the GHBP: proteolytic cleavage of the membrane receptor and a specific mRNA produced by alternative splicing. PMID- 9228078 TI - The murine voltage-dependent anion channel gene family. Conserved structure and function. AB - Voltage-dependent anion channels (VDACs) are pore-forming proteins found in the outer mitochondrial membrane of all eucaryotes. VDACs are the binding sites for several cytosolic enzymes, including the isoforms of hexokinase and glycerol kinase. VDACs have recently been shown to conduct ATP when in the open state, allowing bound kinases preferential access to mitochondrial ATP and providing a possible mechanism for the regulation of adenine nucleotide flux. Two human VDAC cDNAs have been described previously, and we recently reported the isolation of mouse VDAC1 and VDAC2 cDNAs, as well as a third novel VDAC cDNA, designated VDAC3. In this report we describe the structural organization of each mouse VDAC gene and demonstrate that, based on conserved exon/intron boundaries, the three VDAC isoforms belong to a single gene family. The 5'-flanking region of each VDAC gene was shown to have transcription promoter activity by transient expression in cultured cells. The promoter region of each VDAC isoform lacks a canonical TATA box, but all are G+C-rich, a characteristic of housekeeping gene promoters. To examine the conservation of VDAC function, each mouse VDAC was expressed in yeast lacking the endogenous VDAC gene. Both VDAC1 and VDAC2 are able to complement the phenotypic defect associated with the mutant yeast strain. VDAC3, however, is only able to partially complement the mutant phenotype, suggesting an alternative physiologic function for the VDAC3 protein. PMID- 9228077 TI - Engineering the human thyrotropin receptor ectodomain from a non-secreted form to a secreted, highly immunoreactive glycoprotein that neutralizes autoantibodies in Graves' patients' sera. AB - Previous attempts to generate autoantibody-reactive, secreted thyrotropin receptor (TSHR) ectodomain in mammalian cells have failed because of retention within the cell of material with immature carbohydrate. We have overcome this difficulty by performing progressive carboxyl-terminal truncations of the human TSHR ectodomain (418 amino acid residues including signal peptide). Three ectodomain variants (TSHR-261, TSHR-289, and TSHR-309) were truncated at residues 261, 289, and 309, respectively. Unlike the full ectodomain, ectodomain variants were secreted with an efficiency inversely proportional to their size. Secreted ectodomain variants contained approximately 20 kDa of complex carbohydrate. TSHR 261 was chosen for further study because it was secreted very efficiently and neutralized autoantibodies in Graves' patients' sera. This ectodomain variant was partially purified using sequential lectin and nickel-chelate chromatography, permitting the first direct visualization and quantitation of the mammalian TSHR. Most important, very small (nanogram) quantities of this material neutralized 70 100% of TSHR autoantibody activity in all 18 Graves' sera studied. In summary, carboxyl-terminal truncation of the human TSHR ectodomain generates a secreted protein with complex carbohydrate that neutralizes autoantibodies in Graves' patients' sera. Antigenically active TSHR will be valuable for future studies on the diagnosis, pathogenesis, and immunotherapy of Graves' disease. PMID- 9228079 TI - A complex consisting of human replication factor C p40, p37, and p36 subunits is a DNA-dependent ATPase and an intermediate in the assembly of the holoenzyme. AB - Human replication factor C (hRFC) is a multi-subunit protein complex capable of supporting proliferating cell nuclear antigen (PCNA)-dependent DNA synthesis by DNA polymerases delta and epsilon. The hRFC complex consists of five different subunits with apparent molecular masses of 140, 40, 38, 37, and 36 kDa. We have previously reported the expression of a three-subunit core complex, consisting of the p40, p37, and p36 subunits following coupled in vitro transcription translation of the cDNAs encoding these proteins (Uhlmann, F., Cai, J., Flores Rozas, H., Dean, F. B., Finkelstein, J. , O'Donnell, M., and Hurwitz, J. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 6521-6526). Here we describe the isolation of a stable complex composed of the p40, p37, and p36 subunits of hRFC from baculovirus-infected insect cells. The purified p40.p37.p36 complex, like the five-subunit RFC, contained DNA-dependent ATPase activity that was stimulated by PCNA, preferentially bound to primed DNA templates, interacted with PCNA, and was capable of unloading PCNA from singly-nicked circular DNA. In contrast to the five-subunit RFC, the three-subunit core complex did not load PCNA onto DNA. The p40. p37.p36 complex inhibited the elongation of primed DNA templates catalyzed by the DNA polymerase delta holoenzyme. Incubation of the p40.p37.p36 complex with the hRFC p140 and p38 subunits formed the five-subunit hRFC complex that supported PCNA-dependent DNA synthesis by DNA polymerase delta. PMID- 9228080 TI - Glucose transporter isoforms GLUT1 and GLUT3 transport dehydroascorbic acid. AB - Dehydroascorbic acid (DHA) is rapidly taken up by cells and reduced to ascorbic acid (AA). Using the Xenopus laevis oocyte expression system we examined transport of DHA and AA via glucose transporter isoforms GLUT1-5 and SGLT1. The apparent Km of DHA transport via GLUT1 and GLUT3 was 1.1 +/- 0.2 and 1.7 +/- 0.3 mM, respectively. High performance liquid chromatography analysis confirmed 100% reduction of DHA to AA within oocytes. GLUT4 transport of DHA was only 2-4-fold above control and transport kinetics could not be calculated. GLUT2, GLUT5, and SGLT1 did not transport DHA and none of the isoforms transported AA. Radiolabeled sugar transport confirmed transporter function and identity of all cDNA clones was confirmed by restriction fragment mapping. GLUT1 and GLUT3 cDNA were further verified by polymerase chain reaction. DHA transport activity in both GLUT1 and GLUT3 was inhibited by 2-deoxyglucose, D-glucose, and 3-O-methylglucose among other hexoses while fructose and L-glucose showed no inhibition. Inhibition by the endofacial inhibitor, cytochalasin B, was non-competitive and inhibition by the exofacial inhibitor, 4,6-O-ethylidene-alpha-glucose, was competitive. Expressed mutant constructs of GLUT1 and GLUT3 did not transport DHA. DHA and 2 deoxyglucose uptake by Chinese hamster ovary cells overexpressing either GLUT1 or GLUT3 was increased 2-8-fold over control cells. These studies suggest GLUT1 and GLUT3 isoforms are the specific glucose transporter isoforms which mediate DHA transport and subsequent accumulation of AA. PMID- 9228081 TI - Specific activation of retinoic acid receptors (RARs) and retinoid X receptors reveals a unique role for RARgamma in induction of differentiation and apoptosis of S91 melanoma cells. AB - Retinoic acid (RA) and 9-cis-RA induce growth arrest and differentiation of S91 melanoma cells. RA activates retinoic acid receptors (RARs), whereas 9-cis-RA activates both RARs and retinoid X receptors (RXRs). Both classes of receptors function as ligand-dependent transcription factors. S91 melanoma cells contain mRNA for RXRalpha, RXRbeta, RARalpha, RARgamma, and RARbeta in low levels. Among these, only RARbeta gene transcription is induced by retinoids. However, at present the individual role(s) for each RXR and RAR isoform in these processes is unclear. We assessed the function of all isoforms in the S91 melanoma model by using RXR and RAR isoform-specific retinoids to study their effects on cell growth, RARbeta expression, and differentiation. Activation of each of the endogenous RXR or RAR isoforms induces RARbeta gene expression, and blocks cellular proliferation. However, only the RARgamma-ligands cause additional differentiation toward a melanocytic phenotype, which coincides with substantial apoptosis well before morphological changes are apparent. Apoptosis is completely dependent on de novo protein synthesis but cannot be induced by changes in activities of AP-1, protein kinase C, and protein kinase A, nor can it be blocked by the presence of the antioxidant glutathione. These results argue against a specific role for RARbeta, but suggest that RARgamma has a critical role in a genetic switch between melanocytes and melanoma, and induction of ligand dependent apoptosis. PMID- 9228082 TI - Differential regulation of phosphoinositide 3-kinase adapter subunit variants by insulin in human skeletal muscle. AB - The role of phosphoinositide 3-kinase (PI 3-kinase) in insulin signaling was evaluated in human skeletal muscle. Insulin stimulated both antiphosphotyrosine precipitable PI 3-kinase activity and 3-O-methylglucose transport in isolated skeletal muscle (both approximately 2-3-fold). Insulin stimulation of 3-O methylglucose transport was inhibited by the PI 3-kinase inhibitor LY294002 (IC50 = 2.5 microM). The PI 3-kinase adapter subunits were purified from muscle lysates using phosphopeptide beads based on the Tyr-751 region of the platelet-derived growth factor receptor. Immunoblotting of the material adsorbed onto the phosphopeptide beads revealed the presence of p85alpha, p85beta, p55(PIK)/p55gamma, and p50 adapter subunit isoforms. In addition, p85alpha-NSH2 antibodies recognized four adapter subunit variants of 54, 53, 48, and 46 kDa, the latter corresponding to the p50 splice variant. Serial immunoprecipitations demonstrated that these four proteins were associated with a large proportion of the total PI 3-kinase activity immunoprecipitated by p85alpha-NSH2 domain antibodies. Antibodies to p85beta, p55(PIK)/p55gamma, and the p50 adapter subunit also immunoprecipitated PI 3-kinase activity from human muscle lysates. A large proportion of the total cellular pool of the 53-kDa variant, p50, and p55(PIK) was present in antiphosphotyrosine immunoprecipitates from unstimulated muscle, whereas these immunoprecipitates contained only a very small proportion of the cellular pool of p85alpha, p85beta, and the 48-kDa variant. Insulin greatly increased the levels of the 48-kDa variant in antiphosphotyrosine immunoprecipitates and caused smaller -fold increases in the levels of p85alpha, p85beta, and the 53-kDa variant. The levels of p50 and p55(PIK) were not significantly changed. These properties indicate mechanisms by which specificity is achieved in the PI 3-kinase signaling system. PMID- 9228083 TI - Mechanistic studies of the dual phosphorylation of mitogen-activated protein kinase. AB - Previous work on the responses of mitogen-activated protein (MAP) kinase cascade components in a Xenopus oocyte extract system demonstrated that p42 MAP kinase (MAPK) exhibits a sharp, sigmoidal stimulus/response curve, rather than a more typical hyperbolic curve. One plausible explanation for this behavior requires the assumption that MAP kinase kinase (MAPKK) carries out its dual phosphorylation of p42 MAPK by a distributive mechanism, where MAPKK dissociates from MAPK between the first and second phosphorylations, rather than a processive mechanism, where MAPKK carries out both phosphorylations before dissociating. Here we have investigated the mechanism through which a constitutively active form of human MAPKK-1 (denoted MAPKK-1 R4F or MAPKK-1*) phosphorylates Xenopus p42 MAPK in vitro. We found that the amount of monophosphorylated MAPK formed during the phosphorylation reaction exceeded the amount of MAPKK-1* present, which would not be possible if the phosphorylation occurred exclusively by a processive mechanism. The monophosphorylated MAPK was phosphorylated predominantly on tyrosine, but a small proportion was phosphorylated on threonine, indicating that the first phosphorylation is usually, but not invariably, the tyrosine phosphorylation. We also found that the rate at which pulse-labeled monophosphorylated MAPK became bisphosphorylated depended on the MAPKK-1* concentration, behavior that is predicted by the distributive model but incompatible with the processive model. These findings indicate that MAPKK-1* phosphorylates p42 MAPK by a two-collision, distributive mechanism rather than a single-collision, processive mechanism, and provide a mechanistic basis for understanding how MAP kinase can convert graded inputs into switch-like outputs. PMID- 9228084 TI - Differential regulation of adenylyl cyclases by Galphas. AB - Regulation of adenylyl cyclases 1, 2, and 6 by Galphas was studied. All three mammalian adenylyl cyclases were expressed in insect (Sf9 or Hi-5) cells by baculovirus infection. Membranes containing the different adenylyl cyclases were stimulated by varying concentrations of mutant (Q227L) activated Galphas expressed in reticulocyte lysates. Galphas stimulation of AC1 involved a single site and had an apparent Kact of 0.9 nM. Galphas stimulation of AC2 was best explained by a non-interactive two site model with a "high affinity" site at 0.9 nM and a "low affinity" site at 15 nM. Occupancy of the high affinity site appears to be sufficient for Gbetagamma stimulation of AC2. Galphas stimulation of AC6 was also best explained by a two-site model with a high affinity site at 0. 6-0.8 nM and a low affinity site at 8-22 nM; however, in contrast to AC2, only a model that assumed interactions between the two sites best fit the AC6 data. With 100 microM forskolin, Galphas stimulation of all three adenylyl cyclases showed very similar profiles. Galphas stimulation in the presence of forskolin involved a single site with apparent Kact of 0.1-0.4 nM. These observations indicate a conserved mechanism by which forskolin regulates Galphas coupling to the different adenylyl cyclases. However, there are fundamental differences in the mechanism of Galphas stimulation of the different adenylyl cyclases with AC2 and AC6 having multiple interconvertible sites. These mechanistic differences may provide an explanation for the varied responses by different cells and tissues to hormones that elevate cAMP levels. PMID- 9228085 TI - Angiotensin type 2 receptor dephosphorylates Bcl-2 by activating mitogen activated protein kinase phosphatase-1 and induces apoptosis. AB - We examined the cellular and signaling mechanism of angiotensin II (Ang II) type 2 (AT2) receptor-induced apoptosis in PC12W (rat pheochromocytoma cell line) cells that express abundant AT2 receptor but not Ang II type 1 receptor. In these cells, nerve growth factor (NGF) inhibited the internucleosomal DNA fragmentation induced by serum depletion, whereas Ang II antagonized this NGF cell survival action and induced apoptosis. We studied the mechanism of NGF and AT2 receptor interaction on apoptosis by examining their effects on the survival factor Bcl-2. AT2 receptor activation did affect intracellular Bcl-2 protein levels. Bcl-2 phosphorylation was stimulated by NGF, whereas AT2 receptor activation blocked this NGF effect. Pretreatment with antisense oligonucleotide of mitogen-activated protein (MAP) kinase phosphatase-1 enhanced the effects of NGF on MAP kinase activation and Bcl-2 phosphorylation but attenuated the inhibitory effects of AT2 receptor on MAP kinase, Bcl-2 phosphorylation, and apoptosis. Taken together, these results suggest that MAP kinase plays a critical role in inhibiting apoptosis by phosphorylating Bcl-2. The AT2 receptor inhibits MAP kinase activation, resulting in the inactivation of Bcl-2 and the induction of apoptosis. PMID- 9228086 TI - Retinoid X receptor (RXR) ligands activate the human 25-hydroxyvitamin D3-24 hydroxylase promoter via RXR heterodimer binding to two vitamin D-responsive elements and elicit additive effects with 1,25-dihydroxyvitamin D3. AB - We have previously shown that RNA levels of kidney 25-hydroxyvitamin D3-24 hydroxylase (24(OH)ase), a key metabolic enzyme for 1,25-dihydroxyvitamin D3 (1,25(OH)2D3), is up-regulated by retinoids in mice within hours. Deletion analysis of approximately 5500 base pairs of the human 24(OH)ase promoter showed that the sequence between -316 and -142 contained the information necessary and sufficient for retinoid-induced activation of the promoter. This region contains two previously defined vitamin D-responsive elements (VDREs) at -294 to -274 and 174 to -151. Mutation of either VDRE diminished responsiveness of the -316 to -22 promoter sequence to retinoids or 1,25(OH)2D3, while mutation of both VDREs essentially abolished the activity of the ligands via the promoter. Heterologous promoter vectors driven by the VDREs were responsive to a retinoid X receptor (RXR)-selective ligand (LG100268), a retinoic acid receptor (RAR)-selective ligand (TTNPB), or 1,25(OH)2D3, while combinations of LG100268 with either TTNPB or 1,25(OH)2D3 resulted in additive increases in activity. Band shift analyses showed that vitamin D receptor, RAR, or RXR alone did not bind to the VDREs; however, the combination of either vitamin D receptor or RAR with RXR led to retardation of each of the labeled probes. Treatment of nontransfected CV-1 cells with retinoids or 1,25(OH)2D3 resulted in induction of 24(OH)ase RNA, and ligand combinations led to increased RNA levels. These data imply that either or both of the heterodimer partners can be occupied with ligand to induce this enzyme, with dual receptor occupation leading to increased activation. PMID- 9228087 TI - Disruption of microtubules reveals two independent apical targeting mechanisms for G-protein-coupled receptors in polarized renal epithelial cells. AB - G-protein-coupled receptors demonstrate differing trafficking itineraries in polarized Madin-Darby canine kidney (MDCK II) cells. The alpha2A adrenergic receptor (alpha2AAR) is directly delivered to the basolateral subdomain; the A1 adenosine receptor (A1AdoR) is apically enriched in its targeting; and the alpha2BAR subtype is randomly delivered to both domains but selectively retained basolaterally (Keefer, J. R., and Limbird, L. E. (1993) J. Biol. Chem. 268, 11340 11347; Saunders, C., Keefer, J. R., Kennedy, A. P., Wells, J. N., and Limbird, L. E. (1996) J. Biol. Chem. 271, 995-1002; Wozniak, M., and Limbird, L. E. (1996) J. Biol. Chem. 271, 5017-5024). The present studies explore the role of the polarized cytoskeleton in localization of G-protein-coupled receptors in MDCK II cells. Nocodazole or colchicine, which disrupt microtubules, did not perturb lateral localization of alpha2AR subtypes but led to a relocalization the A1AdoR to the basolateral surface, revealed by immunocytochemical and metabolic labeling strategies. Conversely, the apical component of the random delivery of alpha2BAR was not affected by these agents, suggesting microtubule-dependent and independent apical targeting mechanisms for G-protein-coupled receptors in polarized cells. Apparent rerouting of the apically targeted A1AdoR was selective for microtubule-disrupting agents, since cytochalasin D, which disrupts actin polymerization, did not alter A1AdoR or alpha2BAR localization or targeting. These data suggest that multiple apical targeting mechanisms exist for G-protein coupled receptors and that microtubule-disrupting agents serve as tools to probe their different trafficking mechanisms. PMID- 9228088 TI - The O-linked fucose glycosylation pathway. Evidence for protein-specific elongation of o-linked fucose in Chinese hamster ovary cells. AB - O-Linked fucose is an unusual form of glycosylation recently shown to modify the hydroxyls of serine or threonine residues at a strict consensus site within epidermal growth factor-like domains of several serum proteins. Here we demonstrate that Chinese hamster ovary cells modify numerous proteins with O linked fucose and that the fucose is elongated on specific proteins. We have identified at least two forms of O-linked fucose elongation in Chinese hamster ovary cells: a disaccharide (Glcbeta1,3Fuc) and a larger oligosaccharide of indeterminate structure. Interestingly, it appears that the level of monosaccharide accumulates in the cells over time whereas the disaccharide does not. Analysis of the O-linked fucose-containing saccharides on individual proteins revealed that some proteins are modified with the monosaccharide only, whereas others are modified with monosaccharide and disaccharide, or monosaccharide and oligosaccharide. These results suggest that elongation of the O-linked fucose monosaccharide is a protein-specific phenomena. The presence of elongated O-linked fucose moieties suggests that a novel glycosylation pathway exists in mammalian cells with O-linked fucose as the core. PMID- 9228089 TI - Molecular characterization of a small heat shock/alpha-crystallin protein in encysted Artemia embryos. AB - Molecular chaperones protect cells during stress by limiting the denaturation/aggregation of proteins and facilitating their renaturation. In this context, brine shrimp embryos can endure a wide variety of stressful conditions, including temperature extremes, prolonged anoxia, and desiccation, thus encountering shortages of both energy (ATP) and water. How the embryos survive these stresses is the subject of continuing study, a situation true for other organisms facing similar physiological challenges. To approach this question we cloned and sequenced a cDNA for p26, a molecular chaperone specific to oviparous Artemia embryos. p26 is the first representative of the small heat shock/alpha crystallin family from crustaceans to be sequenced, and it possesses the conserved alpha-crystallin domain characteristic of these proteins. The secondary structure of this domain was predicted to consist predominantly of beta-pleated sheet, and it appeared to lack regions of alpha-helix. Unique properties of the nonconserved amino terminus, which showed weak similarity to nucleolins and fibrillarins, are enrichments in both glycine and arginine. The carboxyl-terminal tail is the longest yet reported for a small heat shock/alpha-crystallin protein, and it is hydrophilic, a common attribute of this region. Site-specific differences between amino acids from p26 and other small heat shock/alpha crystallin proteins bring into question the functions proposed for some of these residues. Probing of Southern blots disclosed a multi-gene family for p26, whereas two size classes of p26 mRNA at 0.7 and 1.9 kilobase pairs were seen on Northern blots, the larger probably representing nonprocessed transcripts. Examination of immunofluorescently stained samples with the confocal microscope revealed that a limited portion of intracellular p26 is found in the nuclei of encysted embryos and that it resides within discrete compartments of this organelle. The results in this paper demonstrate clearly that p26 is a novel member of the small heat shock/alpha-crystallin family of proteins. These data, in concert with its restriction to embryos undergoing oviparous development, suggest that p26 functions as a molecular chaperone during exposure to stress, perhaps able to limit protein degradation and thus ensure a ready supply of functional proteins when growth is reinitiated. PMID- 9228091 TI - Carbohydrate binding specificity of the neutrophil-activating protein of Helicobacter pylori. AB - The possible interaction of the neutrophil-activating protein of Helicobacter pylori with target cell glycoconjugates was investigated by the binding of 125I labeled recombinant protein to glycosphingolipids from human neutrophils in solid phase assays. Thereby, a distinct binding of the neutrophil-activating protein to four bands in the acid glycosphingolipid fraction from human neutrophils was detected, whereas no binding to the non-acid glycosphingolipids or polyglycosyl ceramides from these cells was obtained. When using glycosphingolipids not present in the cell membrane of human neutrophils, it was found that the neutrophil-activating protein also bound to sulfated glycosphingolipids as sulfatide and sulfated gangliotetraosyl ceramide. Comparison of the binding preferences of the protein to reference glycosphingolipids from other sources suggested that in human granulocytes, the neutrophil-activating protein of H. pylori preferentially recognizes glycoconjugates with a terminally unsubstituted NeuAcalpha3Galbeta4GlcNAcbeta3Galbeta4GlcNAcbeta sequence. PMID- 9228090 TI - Transforming growth factor-beta regulation of bone morphogenetic protein 1/procollagen C-proteinase and related proteins in fibrogenic cells and keratinocytes. AB - Transforming growth factor-beta1 (TGF-beta1) induces increased extracellular matrix deposition. Bone morphogenetic protein-1 (BMP-1) also plays key roles in regulating vertebrate matrix deposition; it is the procollagen C-proteinase (PCP) that processes procollagen types I-III, and it may also mediate biosynthetic processing of lysyl oxidase and laminin 5. Here we show that BMP-1 is itself up regulated by TGF-beta1 and that secreted BMP-1, induced by TGF-beta1, is either processed to an active form or remains as unprocessed proenzyme, in a cell type dependent manner. In MG-63 osteosacrcoma cells, TGF-beta1 elevated levels of BMP 1 mRNA approximately 7-fold and elevated levels of mRNA for mammalian tolloid (mTld), an alternatively spliced product of the BMP1 gene, to a lesser extent. Induction of RNA was dose- and time-dependent and cycloheximide-inhibitable. Secreted BMP-1 and mTld, induced by TGF-beta1 in MG-63 and other fibrogenic cell cultures, were predominantly in forms in which proregions had been removed to yield activated enzyme. TGF-beta1 treatment also induced procollagen N-proteinase activity in fibrogenic cultures, while expression of the procollagen C-proteinase enhancer (PCPE), a glycoprotein that stimulates PCP activity, was unaffected. In contrast to fibrogenic cells, keratinocytes lacked detectable PCPE under any culture conditions and were induced by TGF-beta1 to secrete BMP-1 and mTld predominantly as unprocessed proenzymes. PMID- 9228092 TI - The human dUTPase gene encodes both nuclear and mitochondrial isoforms. Differential expression of the isoforms and characterization of a cDNA encoding the mitochondrial species. AB - We have previously identified distinct nuclear and mitochondrial isoforms of dUTPase in human cells, reporting the cDNA sequence of the nuclear isoform (DUT N). We now report a cDNA corresponding to the mitochondrial isoform (DUT-M). The DUT-M cDNA contains an 252-amino acid open reading frame, encoding a protein with a predicted Mr of 26,704. The amino-terminal region of the protein contains an arginine-rich, 69-residue mitochondrial targeting presequence that is absent in the mature protein. In vitro transcription and translation of the DUT-M cDNA results in the production of a precursor protein with an apparent molecular mass of 31 kDa as judged by SDS-polyacrylamide gel electrophoresis. The DUT-M precursor is enzymatically active and immunoreacts with a dUTPase-specific monoclonal antibody. Mitochondrial import and processing studies demonstrate that the DUT-M precursor is processed into a 23-kDa protein and imported into mitochondria in vitro. Isoelectric focusing experiments demonstrate that the DUT N has a pI of 6.0, while the processed form of DUT-M has a more basic pI of 8.1, measurements that are in agreement with predicted values. Studies aimed at understanding the expression of these isoforms were performed utilizing quiescent and replicating 34Lu human lung fibroblasts as a model cell culture system. Northern blot analysis, employing an isoform-specific probe, demonstrates that DUT-N and DUT-M are encoded by two distinct mRNA species of 1.1 and 1.4 kilobases, respectively. Western and Northern blot analysis reveal that DUT-M protein and mRNA are expressed in a constitutive fashion, independent of cell cycle phase or proliferation status. In contrast, DUT-N protein and mRNA levels are tightly regulated to coincide with nuclear DNA replication status. Because DUT-N and DUT-M have identical amino acid and cDNA sequences in their overlapping regions, we set out to determine if they were encoded by the same gene. The 5' region of the gene encoding dUTPase was isolated and characterized by a combination of Southern hybridization and DNA sequencing. These analyses demonstrate that the dUTPase isoforms are encoded by the same gene with isoform specific transcripts arising through the use of alternative 5' exons. This finding represents the first example in humans of alternative 5' exon usage to generate differentially expressed nuclear and mitochondrial specific protein isoforms. PMID- 9228093 TI - Residues flanking the HOX YPWM motif contribute to cooperative interactions with PBX. AB - Hox genes encode transcription factors that are major determinants of embryonic patterning. Recently, we and others have shown that specific recognition of target sites in DNA is partly achieved through cooperative interaction with the extradenticle/pre-B-cell transformation-related gene (EXD/PBX) family of homeodomain-containing proteins. This interaction is mediated by the YPWM motif present N-terminal to the homeodomain in HOX proteins. In the present study, we use YPWM peptides to confirm the importance of this motif for mediating HOX/PBX interactions. We also used a novel monoclonal antibody directed against the YPWM to show that occlusion of this motif abrogates cooperativity with PBX. In addition, we present evidence that residues flanking the YPWM, both N-terminally and C-terminally, stabilize the HOX.PBX cooperative complex. Because these flanking residues are also conserved among paralogs, they are likely to help distinguish the specificity of HOX/PBX interactions. Our data further show that the relative importance of individual residues within and flanking the YPWM is dependent on the identity of position 6 of the cooperative binding site (TGATTNATGG). These results suggest that interactions between PBX and the YPWM motif are modified by a base pair predicted to contact the N-terminal arm of the HOX homeodomain. PMID- 9228095 TI - MRI during pregnancy. PMID- 9228094 TI - COX15 codes for a mitochondrial protein essential for the assembly of yeast cytochrome oxidase. AB - The respiratory defect of Saccharomyces cerevisiae mutants assigned to complementation group G4 of a pet strain collection stems from their failure to synthesize cytochrome oxidase. The mutations do not affect expression of either the mitochondrially or nuclearly encoded subunits of the enzyme. The cytochrome oxidase deficiency also does not appear to be related to mitochondrial copper metabolism or heme a biosynthesis. These data suggest that the mutants are likely to be impaired in assembly of the enzyme. A gene designated COX15 has been cloned by transformation of mutants from complementation group G4. This gene is identical to reading frame YER141w on chromosome 5. To facilitate further studies, Cox15p has been expressed as a biotinylated protein. Biotinylated Cox15p fully restores cytochrome oxidase in cox15 mutants, indicating that the carboxyl terminal sequence with biotin does not affect its function. Cox15p is a constituent of the mitochondrial inner membrane and, because of its resistance to proteolysis, probably is largely embedded in the phospholipid bilayer of the membrane. The present studies further emphasize the complexity of cytochrome oxidase assembly and report a new constituent of mitochondria involved in this process. The existence of COX15 homologs in Schizosaccharomyces pombe and Caenorhabditis elegans suggests that it may be widely distributed in eucaryotic organisms. PMID- 9228096 TI - State-of-the-art CT and MRI of the adrenal gland. AB - Both CT and MRI have achieved high accuracy in the investigation of patients suspected of having adrenal pathology. The choice of technique will depend on several factors discussed in the review. The advent of spiral CT has allowed the examination to be tailored to demonstrating the adrenal with very high spatial resolution and it remains the most widely used initial technique. This review concentrates on new techniques for evaluating the incidentally discovered adrenal mass and differentiating between adrenal adenomas and metastases. PMID- 9228097 TI - The role of imaging in adult acute urinary tract infection. AB - Imaging is required in only a minority of patients with urinary tract infection. Some patients who present with severe loin pain are imaged because ureteric colic is suspected. If urinary tract infection does not respond normally to antibiotics, imaging is undertaken to check for evidence of renal obstruction or sepsis. Finally, after the acute infection has been treated, imaging is required in some patients to check for factors pre-disposing to renal damage or to relapsing or recurrent infection. This review discusses the appropriate choice of imaging technique to use in each clinical situation and summarises the expected findings. PMID- 9228098 TI - Bilateral emphysematous pyelonephritis resolving to medical therapy: demonstration by US and CT. AB - Emphysematous pyelonephritis (EPN) is an uncommon and life-threatening necrotizing infection of the renal parenchyma occurring mostly in diabetic patients. It is usually unilateral. Nephrectomy is the current therapeutic procedure. We report the plain radiograph, US and CT findings in a 26-year-old diabetic woman who presented with bilateral EPN and was cured by medical treatment alone. PMID- 9228099 TI - Computed tomography of complications of lung transplantation. AB - In spite of improvements in single or double lung transplantation (LT) technique, complications after LT are not uncommon; the most frequent ale anastomotic complications, infections and rejection (acute or chronic). Early detection of complications of LT allows the optimal therapeutic option to be taken, yielding decreased morbidity and mortality. In some cases, CT plays a key role in early detection of several complications of LT that may not be depicted with other diagnostic modalities, so that knowledge of their CT features is important. In this pictorial review, the authors describe the spectrum of CT features of the complications of LT (including reimplantation response, mechanical problems, acute and chronic rejection, infection, lymphoproliferative disorders, recurrence of the initial disease and complications involving the pleura and the anastomotic sites). In addition, the authors analyze the value of CT compared to that of the other available modalities for the detection of complications of LT. PMID- 9228100 TI - Central airway stenoses: preliminary results of spiral-CT-generated virtual bronchoscopy simulations in 29 patients. AB - The purpose of this study was to determine the feasibility of using virtual bronchoscopy simulations to depict stenoses of the tracheobronchial tree. Virtual bronchoscopy simulations, based on ray casting, were applied to spiral-CT data sets of 29 patients presenting 41 stenoses of the central airways, proved with fiberoptic bronchoscopy. Simulations of the inner walls of the airways were of good quality in 27 of 29 patients. Airway stenoses were depicted in 39 of 41 cases. Evaluation of the length of stenoses and surrounding tissues required simultaneous display of multiplanar reformations. Virtual bronchoscopy provides a valuable road map for bronchoscopy, in an image format familiar to bronchoscopists. PMID- 9228101 TI - Transudative vs exudative pleural effusions: differentiation using Gd-DTPA enhanced MRI. AB - The aim of this study was to investigate the capability of Gd-DTPA-enhanced MRI to differentiate between exudative and transudative pleural effusions. An MRI examination was performed on 22 patients with different types of pleural effusion (10 transudative and 12 exudative effusions). T1-weighted SE images were obtained before and 20 min after administration of Gd-DTPA (0.1 mmol/kg). The degree of enhancement of pleural effusions was evaluated both by visual assessment and by quantitative analysis of images. None of 10 transudative effusions showed significative enhancement, whereas 10 of 12 exudative effusions showed enhancement (sensitivity 83 %, specificity 100 %, positive predictive value 100 %). The postcontrast signal intensity ratios (SIRs) of exudates were significantly higher than corresponding precontrast ratios (P = 0. 0109) and the postcontrast SIRs of exudates were significantly higher than those of transudates (P = 0.0300). Exudative pleural effusions show a significant enhancement following administration of Gd-DTPA. We presume that this may be caused by increased pleural permeability and more rapid passage of a large amount of Gd DTPA from the blood into the pleural fluid in case of exudative effusions. In our limited group of patients, signal enhancement proved the presence of an exudative effusion. Absence of signal enhancement suggests a transudate, but does not exclude an exudate. PMID- 9228102 TI - MR cholangiopancreatography: technique, potential indications, and diagnostic features of benign, postoperative, and malignant conditions. AB - The objective of this article is to review technical aspects, discuss potential clinical indications for MR cholangiopancreatography (MRCP) and demonstrate the spectrum of diagnostic findings in benign, postoperative, and malignant conditions. We describe our current imaging protocol in comparison with other available techniques. Using a non-breath-hold, heavily T2-weighted fast-spin-echo (FSE) sequence with or without respiratory gating we obtained coronal and axial source images and maximum intensity projections (MIPs) in 102 patients with suspected abnormalities of the biliary or pancreatic ducts. Based on this series we demonstrate the diagnostic appearance of a variety of benign, postoperative, and malignant conditions of the biliary and pancreatic ducts and discuss potential clinical indications for MRCP. The non-breath-hold FSE technique enables a consistent image quality even in patients who cannot cooperate well. Respiratory gating increased the rate of diagnostic examinations from 79 to 95 %. Acquisition of coronal and axial source images enables detection of bile duct stones as small as 2 mm, although calculi that are impacted and not surrounded by hyperintense bile may sometimes be difficult to detect. The MIP reconstructions help to determine the level of obstruction in malignant jaundice, delineate anatomical variants and malformations, and to diagnose inflammatory conditions, e. g., sclerosing cholangitis, the Mirizzi syndrome and inflammatory changes in the main pancreatic duct. The MRCP technique also correctly demonstrates the morphology of bilio-enteric or bilio-biliary anastomoses. Because MRCP provides sufficient diagnostic information in a wide range of benign and malignant biliary and pancreatic disorders, it could obviate diagnostic endoscopic retrograde cholangiopancreatography (ERCP) in many clinical settings. The ERCP technique may be increasingly reserved for patients in whom nonsurgical interventional procedures are anticipated. PMID- 9228103 TI - MRI of pancreatic tumors. AB - MRI employing current imaging techniques is able to detect various pancreatic tumors well, and is able to stage and characterize tumors accurately. This review article describes various current MR techniques emphasizing breath hold spoiled gradient echo (SGE) imaging, fat suppression, dynamic gadolinium administration and MR cholangiographic techniques. Pancreatic ductal adenocarcinomas are generally low signal intensity on T1-weighted and T2-weighted images and enhance poorly on immediate post gadolinium SGE images. Islet cell tumors are generally low signal intensity on T1-weighted images, high signal intensity on T2-weighted images and enhance intensely on immediate post gadolinium images. The appearances of less common tumors is also described. PMID- 9228104 TI - The influence of MR field strength on the detection of focal liver lesions with superparamagnetic iron oxide. AB - The purpose of this study was to compare the value of low- vs high-field MR systems in the detection of focal liver lesions after IV administration of iron oxide particles. A prospective study was undertaken which included 20 patients with focal liver lesions on CT or US, or strong clinical suspicion of focal liver disease. Iron oxide particles were administered in an IV drip infusion over 30 min. Magnetic resonance imaging was subsequently performed on a 0.2 and a 1.5-T system. Both examinations were performed in one session. Turbo spin-echo T2 weighted sequences were used for further analysis (at 0.2 T: TR 4050 ms, TE 96 ms; 1.5 T: TR 3000 ms, TE 103 ms). After randomisation, images were analysed by two blinded readers. The evaluation included lesion counts, determination of lesion conspicuity and overall image quality (both graded on a scale 1-5). Quantitative analysis was performed on 29 lesions. Lesion-to-liver signal intensity and contrast-to-noise ratios (CNRs) were calculated. The total lesion count (cumulative counts for two observers) was 59 on the high-field system and 63 on the low-field system. Statistical analysis showed no significant difference. On both systems median value for lesion conspicuity was 3. No statistically significant difference was found. Global image quality was rated higher on the high-field system: 3 vs 2 for the low-field system (p = 0.0017). Quantitative analysis showed no significant difference for lesion-to-liver signal intensity ratios or CNRs. Although subjective image quality is significantly better on the high-field system, this does not result in better lesion detection or better lesion conspicuity. No significant difference in objective quantitative parameters was found in our series. PMID- 9228105 TI - Anatomy of the arterial supply to the liver demonstrated by MRI. AB - The aim of this study was to establish the accuracy of dynamic contrast-enhanced magnetic resonance imaging (DCEMRI) in assessing the site of origin and the patency of the hepatic arteries. Sixty-one patients were examined with serial DCEMRI. MRI was performed at 1.0 T with a rapid multi-section breath-hold fast low-angle shot (FLASH) technique in the coronal oblique plane before and at 10, 40 and 70 s after a bolus of gadolinium-DTPA. The hepatic, left gastric, gastroduodenal, splenic and superior mesenteric arteries were examined. The main portal vein, its right and left intrahepatic divisions, and the splenic and superior mesenteric veins were also assessed. The common hepatic artery was occluded in one patient. The right hepatic artery was seen in 59 patients, left hepatic in 54, left gastric in 43, gastroduodenal in 54, splenic in 60 and superior mesenteric artery in 61. Results were concordant with surgery in 38 of 39 cases and with X-ray angiography in 21 of 22 cases. In the detection of aberrant vessels DCEMRI had a sensitivity of 89 %, a specificity of 100 % and an accuracy of 97 %. All five veins were occluded in 1 patient. The main portal vein was patent in 56 patients, occluded in 2 and narrowed in 2. Thirty-two patients had upper abdominal varices. It is concluded that DCEMRI with sequential imaging provides a non-invasive demonstration of hepatic arterial and venous structures. PMID- 9228106 TI - Pseudolipoma of inverted Meckel's diverticulum: clinical, radiological and pathological correlation. AB - Three cases of isolated inverted Meckel's diverticulum are described. In two cases an initial pathological diagnosis of small bowel lipoma was suggested. In a third case central fat was demonstrated on CT and peristalsis of the intraluminal polypoid mass was observed during US examination. In all three cases small bowel enema examination demonstrated the lesion. Correlation of the clinical, radiological and pathological features is emphasised, as this will allow the correct diagnosis. PMID- 9228107 TI - Brain MRI changes in chronic liver disease. AB - Cirrhotic patients are known to have abnormally high signal principally in the globus pallidus on non-contrast T1-weighted MRI. The purpose of this study was to relate MR changes to clinical and pathological features of chronic liver disease. We confirmed abnormally high signal in the globus pallidus on T1-weighted images in 25 of 28 patients with chronic liver disease, showing that it also occurs in patients who have not yet progressed to cirrhosis. Changes were seen in patients both with and without clinical portosystemic shunting. This abnormality is not responsible for hepatic encephalopathy. Cholestatic disease was more likely to produce marked changes than non-cholestatic disease. No statistically significant correlation was demonstrated between the severity of liver disease and the degree of MR abnormality. However, marked improvement in MR appearances was seen after successful liver transplantation. PMID- 9228108 TI - Solid cerebellar hemangioblastoma with an evolving large cystic component. AB - We present a case of solid cerebellar hemangioblastoma which subsequently developed a large cystic component. PMID- 9228109 TI - Developmental venous anomalies of the posterior fossa with transpontine drainage: report of 3 cases. AB - Developmental venous anomalies (DVA) are considered as variant patterns of cerebral venous drainage. Although generally not rare in the cerebellum, DVA of the brain stem or of the cerebellum with drainage through the brain stem are exceptional findings. Because it is not clear whether DVA may sometimes be of clinical significance, we try to correlate the clinical findings of the patients with the course of the variant vessels. We reviewed the literature and report three additional cases. All patients were examined by MRI and digital subtraction angiography. In particular, we discuss the drainage route as compared with the established patterns of posterior fossa blood drainage, which is directed to the dural sinuses, the petrosal vein or the vein of Galen. In one of our patients suffering from trigeminal neuralgia, the close topical relation of the DVA and the trigeminal nucleus and trigeminal nerve entry zone suggests a causal relationship. In a second case the brain stem symptoms were due to haemorrhage of a concomitant cavernoma. It remains unclear if the occurrence of dysarthria and dysaesthesia in the third patient with brain stem DVA was purely coincidental. The only clinical symptom directly attributable to a DVA with transpontine drainage in our series was trigeminal neuralgia. PMID- 9228110 TI - Direct digital mammography image acquisition. AB - Mammography is a branch of radiology which could benefit greatly from the assimilation of digital imaging technologies. Computerized enhancement techniques could be used to ensure optimum presentation of all clinical images. Beyond this it will facilitate powerful new clinical resources such as computer-assisted diagnosis, tele-mammography, plus digital image management and archiving. An essential precursor to all these advances is the availability of appropriate direct digital mammography (DDM) image-acquisition system(s) to capture high quality breast X-ray image data at the outset. The only practical DDM image acquisition system currently available is (photo-stimulable phosphor) computed radiography. Modern computed mammography (CM) uses similar radiation doses to the patient and produces equivalent, albeit different, image quality to screen-film mammography. Computed mammography offers superior rendition of the skin edge and sub-cutaneous tissue and dense parenchyma, while ensuring equivalent micro calcification detectability. Meanwhile, a variety of new technical approaches to DDM are under active investigation and/or development which promise to supercede film-based mammography. These new (second generation) DDM technologies promise the radiologist superior image quality combined with significant dose savings compared with contemporary imaging systems. In this review we describe and compare the physical and clinical characteristics of CM and the various emerging DDM image-acquisition technologies. PMID- 9228111 TI - Myofibroblastoma of male breast: report of three cases and review of the literature. AB - We report three cases of male breast myofibroblastoma. This uncommon benign tumor arises from breast mesenchyma and is more frequently seen in adult men. Mammographic findings consist of a well-delimited, round to oval dense mass, variable in size but usually 1-4 cm in diameter. No microcalcifications were observed. Ultrasonography confirms the solid nature of the lesion, showing a well circumscribed, homogeneous, hypoechoic mass, compressible with pressure. Although FNA cytology may support the diagnosis, surgical biopsy should be performed. Tumorectomy is the treatment of choice. To our knowledge, no more than 40 cases of breast myofibroblastoma have been reported. This is the first report in the literature which emphasizes the mammographic and ultrasonographic features of this tumor. PMID- 9228112 TI - Percutaneous recanalization of acutely thrombosed vessels by hydrodynamic thrombectomy (Hydrolyser). AB - A hydrodynamic thrombectomy catheter was prospectively evaluated for the treatment of recently thrombosed vessels. A total of 52 consecutive patients (42 males and 10 females; mean age 64 +/- 15 years) presenting with acute or subacute occlusion of dialysis shunts (n = 25), peripheral bypass (n = 14) or native arteries (n = 15) were treated with the Hydrolyser (Cordis Europa NV, Roden, The Netherlands). Mean occlusion time was 4 days (range 1-17 days) and mean thrombus length 19 +/- 11 cm. The Hydrolyser was effective and fast in removing thrombus, regardless of the thrombus length. No major complications were reported. The immediate procedure success rates were 82, 100, 87 and 79 % for Brescia Cimino, dialysis shunt, native arteries and bypass grafts, respectively. Adjunctive thrombolysis (applied for persistence of residual thrombus or thrombosed distal vessels too small for hydrolytic thrombectomy) was required in 4 % of thrombotic dialysis shunts, in 20 % of native arteries and in 50 % of bypass graft occlusions. On angiographic controls, distal embolizations were reported only in native arteries (13 %) and bypasses (14 %); all were successfully treated percutaneously, except for one case treated by Fogarty balloon. Cumulative primary patency rates were respectively at 6 months 56, 62, 78 and 65 % for each indication. We conclude from this preliminary clinical study that hydrodynamic thrombectomy with a Hydrolyser is a promising technique to treat acute occlusions. This device can reduce complications as well as the time required to remove large amounts of thrombus and the use of expensive thrombolytic drugs. PMID- 9228113 TI - The melting pot of "interventional" radiology. PMID- 9228114 TI - Interventional radiology: there is no need for quotation marks. PMID- 9228115 TI - MR of laryngeal and scrotal involvement in multiple symmetrical lipomatosis. AB - Multiple symmetrical lipomatosis is a rare disorder characterized by progressive anomalous deposition of fat typically located in the neck and shoulders. Magnetic resonance imaging allows exact definition of the abnormal fatty tissue and the involvement of deep structures. We describe the MR findings in two patients with the typical fat deposition in the neck and upper thorax which also presented unusual location of abnormal fat. One patient had laryngeal involvement with fatty infiltration of true and false cords. The other patient had inguinal and scrotal large deposition of fat. PMID- 9228116 TI - Clinical radiology in the UK: a time of change. PMID- 9228117 TI - Fibromyalgia--out of control? PMID- 9228118 TI - The fibromyalgia problem. PMID- 9228119 TI - Fibromyalgia: La Maladie est Morte. Vive le Malade! PMID- 9228120 TI - Expression of vascular endothelial growth factor in synovial fibroblasts is induced by hypoxia and interleukin 1beta. AB - OBJECTIVE: To study the mechanism by which hypoxia and inflammatory cytokines mediate angiogenesis in the rheumatoid pannus through their effects on the fibroblast-like type B synoviocyte, the major cell type of normal synovia. METHODS: Fibroblasts were prepared from synovial tissue of healthy and diseased individuals, and cultured in the presence of various stimuli. The expression of vascular endothelial growth factor (VEGF) was assessed by ELISA and reverse transcription polymerase chain reaction. RESULTS: Unlike normal fibroblasts, synovial fibroblasts from rheumatoid arthritis (RA) and osteoarthritis constitutively secreted significant levels of VEGF, which is known to act directly on endothelial cells. VEGF secretion was further inducible by both hypoxia and interleukin 1beta (IL-1beta) and these increases were additive. In contrast, tumor necrosis factor alpha was unable to induce VEGF expression. CONCLUSION: Under hypoxia or IL-1 stimulation, conditions common to the inflamed synovium, type B synoviocytes secrete increased levels of VEGF, which is likely to act on nearby endothelia, promoting angiogenesis. The constitutive expression of VEGF in rheumatoid synovial fibroblasts may reflect an altered phenotype involved in the pathology of RA. PMID- 9228121 TI - Detection of rubella, mumps, and measles virus genomic RNA in cells from synovial fluid and peripheral blood in early rheumatoid arthritis. AB - OBJECTIVE: To determine, by studying patients with early rheumatoid arthritis (RA), whether rubella virus (rubella), mumps virus (mumps), or measles virus (measles) plays a role in the pathogenesis of RA. METHODS: Polymerase chain reaction combined with reverse transcription was used to detect viral RNA in peripheral blood mononuclear cells (PBMC) or synovial fluid (SF) cells. The patients with RA had newly diagnosed disease (duration < or = 1 year). The controls were patients with other arthropathies. RESULTS: Rubella genomic RNA was not detected in SF cells from patients with early RA or from controls, or in PBMC from patients with RA. It was found in PBMC of one of 46 patients with other arthropathies. Mumps or measles genomic RNA was detected in PBMC samples from 1.8% (1/54) and 9.3% (5/54) of RA, respectively, and from 4.3% (2/46) and 6.5% (3/46) of control patients. The SF cell samples harbored mumps or measles RNA in 4.8% (2/42) and 7.1% (3/42) of patients with RA, respectively; the corresponding value was 6.5% (2/31) for control patients, for both mumps and measles. CONCLUSION: Our findings suggest rubella, mumps, or measles do not play a role in the etiopathogenesis of RA. PMID- 9228122 TI - Variation among rheumatologists in clinical outcomes and frequency of office visits for rheumatoid arthritis. AB - OBJECTIVE: To estimate the variation among rheumatologists in clinical outcomes and frequency of office visits for patients with rheumatoid arthritis (RA), after accounting for patient demographic and clinical characteristics and treatments prescribed. METHODS: Multiple regression analysis using random effects for rheumatologists and adjustments for patient characteristics and treatments received, based on data derived from a panel study of persons with RA. RESULTS: During the years 1984-1993, rheumatologists accounted for a moderate amount of the total variation in clinical outcomes and nearly one-third of the total variation in frequency of office visits. For example, in 1993 rheumatologist associated variation in 4 clinical outcomes ranged from 16 to 25%, while the variation in office visit frequency attributable to rheumatologists stood at 46% of the total variation. However, rheumatologist associated variation in clinical outcomes was not statistically significant in any year, while variation in office visits was highly significant in all years (p < or = 0.0001). Although there was an increase in the percentage of variation attributable to rheumatologists for all outcomes examined across the years of this study, the time trend reached statistical significance only for frequency of office visits (2.4% per year; p = 0.0135) and functional status (1.6% per year; p = 0.0034). CONCLUSION: The magnitude and strength of rheumatologist associated variation in frequency of office visits, without comparable strength in the variation in clinical outcomes, may suggest inefficiencies in the use of resources for the care of persons with RA. Further work is needed to directly examine the relationship between health outcomes and resource utilization. PMID- 9228123 TI - Role of HLA-DR-DR and DR-DQ associations in the expression of extraarticular manifestations and rheumatoid factor in rheumatoid arthritis. AB - OBJECTIVE: To investigate the role of HLA-DRB1 genotypes and HLA-DRB1*0401 DQB1*03 haplotypes in the expression of extraarticular manifestations and rheumatoid factor (RF) in rheumatoid arthritis (RA). METHODS: 189 patients with RA were classified according to the presence of vasculitis and seropositivity for RF. IgM RF were determined by at least 2 of the following methods: standard latex agglutination, the Waaler-Rose test, and nephelometry. HLA genotyping was performed by reverse dot blot hybridization for DRB1 alleles and the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for DQB1 alleles. RESULTS: The QKRAA susceptibility sequence, which characterizes the HLA-DRB1*0401 allele, was observed in 71.5% of the 21 patients with vasculitis and 57.6% of the 158 RF positive patients. The influence of a 2nd allele within the major histocompatibility complex was observed but the allele differed according to the clinical features examined. Higher risk for vasculitis was observed in patients who carried 2 DRB1 susceptibility alleles, one characterized by the QKRAA sequence and the other by the QRRAA sequence (OR = 3). Conversely, the higher risk for IgM RF positivity was observed in patients who carried the QKRAA sequence of the DRB1 alleles with the DQB1*0301 alleles of the DQ region (OR = 4.7). CONCLUSION: Our data suggest that a distinct immunogenetic association is involved in the extraarticular manifestations of RA and in the expression of RF. PMID- 9228124 TI - Reinstitution of gold after gold induced proteinuria. AB - OBJECTIVE: To assess the benefits and risks to patients who resumed gold after discontinuation of gold because of proteinuria. METHODS: We conducted a review of records of all patients who had resumed gold after discontinuing treatment because of proteinuria. RESULTS: Eight patients developed proteinuria > or = 500 mg/dl while receiving 50 mg weekly doses in the first year of treatment. Proteinuria took a minimum of 4 mo to resolve. No patient had a recurrence of proteinuria when gold was resumed at a lower dosage of 25 mg every 1-2 weeks. This strategy has been successful in 5/8 patients who continue taking gold and are markedly improved compared to pretreatment status after 3-7 years (mean 5 years) of followup. CONCLUSION: The risk of gold induced proteinuria may be dose related and reinstitution of gold at a lower dose is safe in our experience and effective in selected patients. PMID- 9228125 TI - Progression of cervical spine changes in patients with early rheumatoid arthritis. AB - OBJECTIVE: To evaluate the development and progression of radiological changes of cervical spine in early rheumatoid arthritis (RA). METHODS: Sixty-seven patients with early RA treated actively with disease modifying antirheumatic drugs were followed prospectively for a mean of 6.5 years. Conventional clinical and laboratory variables were used for measuring disease activity and radiographs of the cervical spine, hands, and feet were taken serially during the followup. RESULTS: Thirty percent (20/67) of the patients showed radiological evidence of the cervical spine involvement characteristic of RA. Atlantoaxial subluxation was the first type of cervical change to occur, followed by vertical and subaxial subluxations and erosions. Patients with cervical involvement were initially more often rheumatoid factor positive and had higher initial C-reactive protein level than patients without cervical changes. Also, radiological progression of peripheral joints was associated with cervical involvement. HLA-DR4 or B27 positivity did not seem to influence early involvement of cervical spine. CONCLUSION: Involvement of the cervical spine begins early in RA. Therefore, cervical radiographs should be included in the clinical evaluation during the first years of disease onset, especially in patients with rapid radiological progression in peripheral joints. Aggressive therapy is emphasized in these patients, including the conservative treatment of cervical spine. PMID- 9228126 TI - Continuous progression of radiological destruction in seropositive rheumatoid arthritis. AB - OBJECTIVE: To examine the radiographic changes in rheumatoid arthritis (RA) occurring over a 20 year period. METHODS: In 103 patients with recent (< 6 months) seropositive RA, radiographs were taken at onset and at 1, 3, 8, 15, and 20 years from entry. Larsen grades for wrist and subtalar joints were first multiplied by 5, and these together with Larsen grades for the 1st to 5th metacarpophalangeal (MCP) joints, the 1st interphalangeal, and the 2nd to 5th metatarsophalangeal (MTP) joints of the feet (24 joints) were added to form a Larsen score of 0-200. Larsen grades for the 1st to 5th MCP, wrists, and 2nd to 5th MTP (20 joints) were added to form a Larsen score of 0-100. RESULTS: The means of the 0-200 Larsen score were 4.3, 12.9, 26.9, 55.7, 77.5, and 86.4, and of the 0-100 Larsen score 2.5, 7.0, 13.9, 28.2, 39.4, and 44.5. CONCLUSION: Seropositive RA is a chronic disease still leading to continuous progression of joint damage 20 years after onset. PMID- 9228127 TI - Reference curves of radiographic damage in patients with rheumatoid arthritis: application of quantile regression and fractional polynomials. AB - OBJECTIVE: To (1) introduce the methodology of quantile regression and fractional polynomials; (2) test the application of this methodology to develop, conditional on disease duration, preliminary reference curves of radiographic damage in patients with rheumatoid arthritis (RA); and (3) prove the importance of the definition and selection of the reference group when developing reference curves. METHODS: The study design was cross sectional. The main study factors were disease duration and radiographic damage using the Larsen score. The 2 study samples were 98 patients from a multicenter trial of cyclosporine and 203 patients with RA from a teaching hospital clinic. RESULTS: Using disease duration as the time dependent covariate we constructed quantile regression reference curves of radiographic damage. The reference curves for the 2 samples differed in shape, location, and slope. CONCLUSION: Quantile regression and fractional polynomials simplify the construction of reference curves when data cannot be easily modified to meet assumptions of normality, linearity, and constant variance. Quantile reference curves provide clinicians with a useful clinical tool to measure outcome at arbitrary timepoints, to interpret change, and to set treatment objectives. However, the definition and selection of the reference used to construct the reference curves is of critical importance. PMID- 9228128 TI - Radiographic results from the Minocycline in Rheumatoid Arthritis (MIRA) Trial. AB - OBJECTIVE: To assess radiographically determined disease progression in patients in the Minocycline in Rheumatoid Arthritis (MIRA) Trial. METHODS: A double blind, randomized, multicenter, 48 week trial of oral minocycline (200 mg/day) or placebo in 6 clinical centers in the United States. Patients include 219 adults with active RA previously receiving limited treatment with disease modifying drugs. Posteroanterior films of the hands from baseline and final visits, blinded for sequence, were read for erosions and joint space narrowing by trained observers. Outcomes included rate of disease progression (change/month) and percentage of patients with progression from baseline, newly involved joints, and newly erosive disease. RESULTS: Using intent-to-treat analyses, progression rates for erosions (0.11 +/- 0.42 minocycline, 0.17 +/- 0.41 placebo; p = 0.47) and joint space narrowing (0.16 +/- 0.55 minocycline and 0.23 +/- 0.71 placebo; p = 0.14) were similar. (Power 43% to detect a 50% difference.) Newly erosive joints occurred more frequently in the placebo group (44 vs 32%; p = 0.08), not a statistically significant difference. CONCLUSION: Radiographic measurement of disease progression using 4 measures failed to show a significant difference between minocycline and placebo treatment, although for all methods there was a trend toward treatment benefit, consistent with reported clinical results. A one year trial duration, high measurement variability, and slow rate of radiographic progression in this cohort may explain the low power to detect a treatment effect. The measurement that denoted "newly involved" joints was most sensitive in detecting change. In future trials longer term assessment (minimum 2 years) of radiographic changes and further comparison of measures of disease progression are warranted. PMID- 9228129 TI - Magnetic resonance imaging of the wrist in defining remission of rheumatoid arthritis. AB - OBJECTIVE: To assess the efficacy of magnetic resonance imaging (MRI) in objectively defining a state of remission in rheumatoid arthritis (RA) after treatment. METHODS: Ten patients with RA involving the wrist were evaluated before treatment with methotrexate and hydroxychloroquine, and then mean 14 mo later with a followup evaluation. Clinical variables, laboratory measurements, and MRI using various techniques (T1 weighted image, T2 weighted image, fat suppression T2 weighted image, postcontrast T1 weighted image, postcontrast dynamic image, postcontrast 3 dimensional image) were observed. Remission was defined by ACR criteria. MRI changes were observed using 3 variables: extent of synovial proliferation; extent of bone marrow edema; and development of new erosion. In 6 of 10 patients, synovial signal intensity time curve changes at 30 s (E30 ratio) were determined for quantitative assessment of synovitis. RESULTS: Four patients achieved remission and 6 did not. All patients in remission showed decrease in extent of synovial proliferation and bone marrow edema with no newly developed erosion after treatment, compared to baseline. Five of 6 patients in nonremission showed newly developed erosions with variable changes in extent of synovial proliferation and bone marrow edema. E30 ratio was determined in 3 patients in the remission group and 3 in the nonremission group, with 48% reduction in the former compared to 9% reduction in the latter. CONCLUSION: MRI is feasible for objectively defining remission and assessing the therapeutic effect of antirheumatic drugs; utility of MRI measures in clinical remission criteria remains to be verified. PMID- 9228130 TI - The association between external weather conditions and pain and stiffness in women with rheumatoid arthritis. AB - OBJECTIVE: To determine the self-reported prevalence of weather sensitivity in a sample of female patients with rheumatoid arthritis (RA), and to determine if there is objective evidence of associations between weather and pain and stiffness in female patients with RA. METHODS: Fifty-three female patients residing in the Sydney metropolitan area participated in a study on the psychological determinants of disability from 1985 to 1987. During the study, subjects recorded pain on a visual analog scale and duration of morning stiffness for 14 day periods at 3-4 monthly intervals over 1-3 years (X = 15.7 months). After completion of the study, data on weather conditions were collected from the Bureau of Meteorology for the days that pain and stiffness records were made. Descriptive statistics and autoregression were used to analyze the data. RESULTS: Sixty percent of subjects reported that they were sensitive to weather. Six weather variables made a statistically significant contribution to daily pain score (p < 0.0001). However, they accounted for only 2.5% of the variance. Two weather variables contributed to duration of morning stiffness (p < 0.0001), but again these variables accounted for only a small portion of the variance (1.1%). A separate analysis for pain was carried out on the data from subjects who reported being weather sensitive. The results were consistent with those of the other analyses, with 2 variables accounting for only 1.7% of the variance (p < 0.0001). CONCLUSION: On the basis of these results it appears that weather makes only a minimal contribution to pain and stiffness in women with RA. The study may have been limited by its use of static measures of weather variables and pain. Further research using dynamic measures of pain and weather and a more extensive range of weather variables is needed. PMID- 9228131 TI - Short and longterm outcomes for patients with systemic rheumatic diseases admitted to intensive care units: a prognostic study of 181 patients. AB - OBJECTIVE: To determine short and longterm outcomes and prognostic factors for patients with systemic rheumatic diseases admitted to intensive care units in 4 teaching hospitals. METHODS: All adult intensive care unit admissions over a 12 year period for systemic rheumatic diseases were retrospectively assessed. One hundred and eighty-one patients with a mean age of 57 +/- 17 years were studied. RESULTS: The death rate in intensive care units was 33% (59/181) and in-hospital mortality was 43% (77/181). One hundred and four patients were discharged alive from hospital; 40 died during followup (mean 105 +/- 7 mo). The estimated 5 year survival rate for the discharged patients was 69%. The 4 factors significantly associated with in-hospital mortality by multivariate analysis were simplified acute physiologic score (p = 10(-4)), poor prior health status (p = 10(-4)), corticosteroid administration (p = 0.005), and the reason for admission; mortality was higher in the group admitted to intensive care for infectious complication (55 versus 34% for others; p = 0.006). In contrast, in-hospital mortality was not influenced by age or by systemic rheumatic diseases. Using Cox's model, only age over 60 years was a prognostic factor significantly associated with an increase in longterm mortality (p = 10(-4)). CONCLUSION: The short term outcome for patients with systemic rheumatic diseases in intensive care units was poor. The longterm prognosis after hospital discharge appeared fair, although the standardized mortality ratio was 5-fold that of a nonselected population. Short and longterm prognoses were similar for different systemic rheumatic disease groups. PMID- 9228132 TI - Administration of systemic matrix metalloproteinase inhibitors maintains bone mechanical integrity in adjuvant arthritis. AB - OBJECTIVE: To evaluate the influence of systemic tetracycline derived antimetalloproteinase compounds on bone morphology and mechanical integrity. METHODS: Male Lewis rats (n = 78) were randomly assigned to one of 10 groups, comprising controls, adjuvant arthritis (AA), and adjuvant arthritis with various combinations of 2 chemically modified, non-antimicrobial tetracycline derivatives (CMT3 or CMT8) with either of 2 nonsteroidal antiinflammatory agents (flurbiprofen or tenidap). After AA induction (23 days), pharmacological efficacy was assessed by inflammatory indices, body mass changes, joint radiological destruction scores, and pyridinoline collagen derived crosslinks. The structural and material properties of the rat femoral neck were assessed biomechanically. RESULTS: Neither CMT had an antiinflammatory effect, but flurbiprofen and tenidap (alone or together with either CMT) significantly reduced joint inflammation. Pyridinoline excretion increased markedly in untreated AA, but was substantially normalized by either CMT3 alone or by CMT8 with flurbiprofen. AA produced significant deleterious effects on femoral neck structure and mechanical properties. Administration of either CMT, however, had positive effects on the amount of bone and the biomechanical properties of rat femoral neck, but not the mineralization of the bone in the rat femoral neck. CONCLUSION: These data suggest that tetracycline derived antimetalloproteinase compounds can significantly and positively influence bone mechanical integrity associated with inhibition of collagen breakdown. PMID- 9228133 TI - Changes of neutrophil migration without modification of in vitro metabolism and adhesion in Behcet's disease. AB - OBJECTIVE: Increase of neutrophil chemotaxis in Behcet's disease (BD) has been described, but it is not clear whether there is a correlation with other variables of neutrophil function and whether these modifications correlate with disease activity. METHODS: We studied neutrophil functions in patients with BD in the acute phase in comparison with healthy control subjects and with the same patients during disease remission, with or without therapy. We investigated in vivo neutrophil migration by Senn's skin window technique and measured adhesion assay and superoxide production in circulating and migrating neutrophils after different stimuli. RESULTS: Neutrophil migration in vivo was 101.3 +/- 17.9 x 10(6) polymorphonuclear lymphocytes (PMN)/cm2/24 h in patients with BD in the acute phase and 66.1 +/- 7.8 x 10(6) PMN/cm2/24 h in controls (p < 0.001). No correlation was found between leukocyte counts and neutrophil migration. Neutrophil migration evaluated in the same patients in a phase of disease remission was 58.3 +/- 10.3 x 10(6) PMN/cm2/24 h (p < 0.001 vs acute phase, not significant vs controls). The neutrophils of the exudate were normally primed to response to the chemotactic peptide fMLP. No differences between the 2 groups were found in superoxide production, adhesion under basal conditions, or in response to different stimuli by circulating and migrating neutrophils. CONCLUSION: Abnormally high migration of neutrophils in the active phase of BD is the only consistent neutrophil dysfunction. Since this modification is reversed by therapy, the evaluation of in vivo neutrophil migration may be useful in diagnosing and monitoring disease activity. Blood neutrophils have normal responses to different stimuli, indicating they are not primed by the disease state. PMID- 9228134 TI - Radiographic hand osteoarthritis: incidence, patterns, and influence of pre existing disease in a population based sample. AB - OBJECTIVE: Osteoarthritis (OA) is the most common type of arthritis; involvement of joints in the hand is highly prevalent, especially in the elderly. Few data are available on the incidence of hand OA in men and women or on the association between OA in one hand joint with incidence in others. METHODS: We studied the cumulative incidence of radiographic hand OA in a population based group of men and women, and evaluated whether baseline OA in one joint affected OA rates in other joints in the hand. Study subjects were 751 members of the Framingham Study cohort, who had a baseline right hand radiograph taken in 1967-1969 (mean age 55+/-5.58) and followup radiographs 24 years later. RESULTS: In those without OA at baseline, women had more incident disease than men in almost all hand joints, but the joints most frequently affected were the same in both sexes: the distal interphalangeal (DIP), followed by the base of the thumb, proximal interphalangeal (PIP), and metacarpophalangeal (MCP) joints. The MCP joint group was the only one in which the incidence in men was comparable to incidence in women. Prevalent OA in one or more joints in a row (e.g., MCP) markedly increased the risk of incident OA in other joints in the same row. Also, prevalent OA in one joint in a finger (a ray) increased the risk of incident OA in other joints in that ray. Prevalent OA in either DIP or PIP joints at baseline substantially increased the risk of incident OA in all other hand joints. Thumb base OA at baseline increased risk in MCP joints, and to a lesser extent, DIP and PIP joints. CONCLUSION: Cumulative incidence was generally higher in women than men, baseline OA in one joint in a row markedly increased the risk of developing OA in other joints in the same row, and baseline OA in a joint in a ray similarly increased risk in that ray. Interphalangeal joint OA at baseline appeared to increase subsequent OA in all hand joints, baseline OA in the thumb was not as strong a predictor. This descriptive information on incidence of radiographic hand OA should generate new hypotheses about why OA affects hands in particular patterns. PMID- 9228135 TI - Knee pain and knee osteoarthritis severity in self-reported task specific disability: the Johnston County Osteoarthritis Project. AB - OBJECTIVE: To evaluate the contributions of radiographic knee osteoarthritis (OA) and knee pain severity to self-reported disability performing upper and lower extremity tasks in a rural, population based sample. METHODS: Data from 1192 African-American and Caucasian participants in the Johnston County Osteoarthritis Project were analyzed with multiple logistic regression to examine the roles of Kellgren-Lawrence radiographic knee OA grade and knee pain severity in self reported difficulty performing 20 activities of the Health Assessment Questionnaire. Potential confounders included age, sex, race, marital status, education, and body mass index. RESULTS: Forty-three percent reported difficulty performing at least one task. Mild knee pain was independently associated with difficulty performing 16 upper and lower extremity tasks, and moderate/severe knee pain with all 20 tasks, with little change after adjustment (p < 0.0001). In contrast, mild radiographic knee OA was associated with difficulty in only 4 mobility and transfer tasks: climbing 5 steps, taking a tub bath, getting in/out of a car, and performing chores. Moderate/severe radiographic knee OA was associated with difficulty in 17 of 20 tasks (in 10 of 17, p < 0.0001), except lifting a cup, opening car doors, and turning faucets. However, no associations between radiographic knee OA and difficulty were statistically significant after adjustment for knee pain and the above factors. CONCLUSION: Knee pain severity was the strongest risk factor for self-reported difficulty performing tasks of upper and lower extremity function. Future studies of disability should include data on knee pain severity. PMID- 9228136 TI - Cartilage damage as a result of hemarthrosis in a human in vitro model. AB - OBJECTIVE: To investigate the direct effect of blood and blood components on human cartilage in vitro. METHODS: Healthy human articular cartilage tissue was obtained post mortem and cultured according to standard procedures. The harmful effects of whole blood and various isolated blood components as well as the reversibility of these effects were assessed by means of proteoglycan synthesis and proteoglycan release. RESULTS: Whole blood anticoagulated with heparin, coagulated blood, mononuclear cells (MNC), erythrocytes [red blood cells (RBC)] and plasma, in this order of potency, decreased proteoglycan synthesis in a dose dependent manner. The effect of the combination of MNC and RBC in concentrations equivalent to those in whole blood was significantly greater than the effect of each of these isolated components alone and did not differ from that of whole blood. Moreover, cartilage exposed for 4 days to this combination exhibited irreversible inhibition of proteoglycan synthesis. The effect was similar to that of whole blood, the opposite of that of the individual components or other combinations. CONCLUSION: Results suggest a direct irreversible harmful effect of whole blood on cartilage, whereby MNC and RBC together are the main factors. Taking into account that the concentration of blood during hemarthrosis approaches 100% and the natural evacuation time of blood from a joint is about 4 days, our results suggest that prompt evacuation of intraarticular blood or prevention of intraarticular bleeding might be crucial in preventing cartilage damage. PMID- 9228137 TI - Expression of N-acetyl-D-galactosamine associated epitope in synovium: a potential marker of glycoprotein production. AB - OBJECTIVE: To investigate synovial glycoprotein production in situ, a novel monoclonal antibody (Mab), A13D8, was used to evaluate the expression of an epitope containing N-acetyl-D-galactosamine (GalNAc) in normal and pathological synovium. METHODS: Immunohistological and cytochemical analysis of synovial tissue samples was undertaken with single and double staining techniques using the A13D8 Mab, anti-CD68, vascular cell adhesion molecule-1 (VCAM-1), the hyaluronan associated enzyme uridine diphosphoglucose dehydrogenase (UDPGD), and the anti-Golgi Mab SSN/HR-1992. The specificity of the A13D8 Mab was established through blocking studies using carbohydrate residues, including GalNAc and N acetylglucosamine (GlcNAc). RESULTS: A13D8 is expressed intensely in the cytoplasm of normal type B lining cells, which coexpress VCAM-1 and UDPGD, and is not expressed by CD68+ type A lining cells. In the lining layer of RA synovium, there is a negative correlation between A13D8 expression and the level of lymphocytic infiltration in the sublining areas (r = -0.43, p < 0.001). The endothelium of a subset of venules, typically in lymphocyte-rich aggregates, also stains intensely for A13D8. Pretreatment of the Mab with GalNAc completely eliminates the tissue staining, as well as the 110 kDa band seen on immunoblot, whereas pretreatment of A13D8 with GlcNAc and lactose has no effect. Double staining of HEp-2 cells with A13D8 and the anti-Golgi Mab SSN/HR-1992 reveals co localization of the A13D8 epitope to the Golgi apparatus. CONCLUSION: Type B synovial lining cells and selected synovial endothelium express GalNAc containing epitope identified by Mab A13D8. Marked reduction in the expression of this epitope in the lining layer of inflamed RA synovium suggests that the synovial production of GalNAc containing glycoproteins, such as mucins, may be altered in this disorder. PMID- 9228138 TI - High prevalence of gout and related risk factors in Taiwan's Aborigines. AB - OBJECTIVE: To estimate the prevalence of gout and to examine its risk factors among Taiwan's Aborigines compared with non-Aborigines. METHODS: Data were collected from persons older than 40 years living in 3 aboriginal and 2 non aboriginal districts in Taiwan by a community survey. Cases of gout were identified from self-reporting of a doctor's diagnosis based on clinical criteria. Baseline variables and biochemical data were examined as risk factors for the development of gout. RESULTS: The prevalence of gout history was found to be 15.2% (25/165) and 4.8% (11/231) among aboriginal men and women, respectively, compared with a prevalence rate of 0.3% among non-Aborigines. A logistic regression model showed that aboriginal men older than 60 years with hyperuricemia were more severely affected by gout than any other group (p < 0.05). CONCLUSION: A high prevalence of gout among Taiwanese Aborigines was observed in this study and race was the most significant risk factor associated with the disease. PMID- 9228139 TI - Reliability, validity, and sensitivity of a Swedish version of the revised and expanded Arthritis Impact Measurement Scales (AIMS2). AB - OBJECTIVE: To evaluate the reliability, validity, and sensitivity of a Swedish version of the Arthritis Impact Measurement Scales (AIMS2) in patients with rheumatoid arthritis (RA). METHODS: Reliability was assessed by a test-retest procedure with a 3-week interval and Cronbach's coefficient of internal consistency. Convergent validity was evaluated by correlation coefficients with the Health Assessment Questionnaire (HAQ), Mood Adjective Checklist (MACL), and disease activity variables. The sensitivity to change was assessed in 40 patients treated with disease modifying antirheumatic drugs, 24 mo after the start of the study. RESULTS: Significant differences were found for the Walking and Bending, Arthritis Pain, and Work scales (p < 0.05) in the test-retest reliability study. Internal consistency coefficients for the AIMS2 scales were 0.68-0.91. Convergent validity was established by significant correlations between the different scales in AIMS2 and the HAQ, MACL, and disease activity variables. Sensitivity to change was observed in 5 of 11 scales in AIMS2. CONCLUSION: After modifying some reliability problems due to a more differentiated scale and cultural differences between the Swedish and American patients with RA, the reliability, validity, and sensitivity to change of AIMS2 were found satisfactory. PMID- 9228140 TI - Patient education in arthritis: randomized controlled trial of a mail-delivered program. AB - OBJECTIVE: Self-management courses in arthritis have been shown to improve outcomes and to decrease medical resource utilization. We studied the effectiveness of a mail-delivered arthritis self-management program with the potential for extending these effects more broadly. METHODS: Randomized controlled trial of 375 program participants and 434 controls over a 6 month period. Baseline and 6 month data were analyzed for each group and between groups by paired 2 sample t test. The intervention consists of health assessment questionnaires at 3 month intervals, with computer processed recommendation letters and reports individualized to age, diagnosis, education level, disability, pain, medication, and other patient-specific variables. RESULTS: At 6 months, outcomes of function (4.7%; 95% confidence limits 2.7, 6.7), decreased pain (9%; 2.8, 15.2), global vitality (7%; 2.8, 11.2), and joint count (28%; 20, 36) were improved in the program group compared with controls (p < 0.01). Exercise (35%; 26, 44) and self-efficacy (14.7%; 9, 20) were increased in the program group but not controls (p < 0.001). Doctor visits/year were decreased by 16% (3, 29) in the program group compared with controls (p < 0.05) and days missed work or confined to home decreased by 52% (-3, 107) in the program group compared with controls (p = 0.06). At one year, patients in the original program group continued to improve, and the control group, provided with the program in the second 6 months, showed improvement similar to the first 6 months of the original program group. CONCLUSION: A mail-delivered arthritis self-management program can positively affect patient outcomes and can decrease medical resource utilization. PMID- 9228142 TI - Ophthalmological monitoring of patients taking antimalarials: preferred practice patterns. PMID- 9228141 TI - Hypothalamic-pituitary-insulin-like growth factor-I axis dysfunction in patients with fibromyalgia. AB - OBJECTIVE: To investigate the serum levels of insulin-like growth factor-I (IGF I) in patients with fibromyalgia (FM) compared to healthy controls and patients with other rheumatic diseases, and to explore possible etiologic mechanisms of low IGF-I levels in patients with FM. METHODS: Five hundred patients with FM and 152 controls (74 healthy blood donors, 26 myofascial pain patients and 52 patients with other rheumatic diseases) were studied. All had measurements of acid extracted serum IGF-I. A subset of 90 patients with FM were evaluated for clinical features that might explain low IGF-I levels. Twenty-five patients with FM underwent growth hormone (GH) provocation testing with l-dopa and clonidine. RESULTS: The mean serum IGF-I level in patients with FM was 138 +/- 56 ng/ml and in controls 215 +/- 86 ng/ml (p = 0.00000000001). Low levels of IGF-I were not due to depression, tricyclic medications, nonsteroidal antiinflammatory drugs, poor aerobic conditioning, obesity, or pain level. Patients with focal myofascial pain syndromes had normal IGF-I levels (236 +/- 68), as did most patients with other rheumatic disorders, unless they had concomitant FM. Patients with FM with initially normal levels often had a rapid decline of IGF-I over 1 to 2 years. Most patients with FM with low IGF-I levels failed to secrete GH after stimulation with clonidine and l-dopa. CONCLUSION: Many, but not all, patients with FM have low levels of IGF-I that cannot be explained by clinical associations. These results suggest that low IGF-I levels in patients with FM are a secondary phenomenon due to hypothalamic-pituitary-GH axis dysfunction. PMID- 9228143 TI - Antimalarial workshop. PMID- 9228144 TI - Evolution of the T cell receptor beta repertoire from synovial fluid T cells of patients with juvenile onset rheumatoid arthritis. AB - OBJECTIVE: To determine the level of T cell clonal expansion and the proportion of T cells that persist over time in the synovial fluid (SF) of patients with juvenile onset rheumatoid arthritis (JRA). METHODS: We collected SF samples from each of 3 patients with JRA at 2 to 3 year intervals. To measure expression across the entire spectrum of Vbeta families in each of 7 fluids examined, we synthesized and amplified dscDNA from all 24 Vbeta families with a single reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: The proportion of clonally expanded T cells and persistent T cells is low and variable among patients. CONCLUSION: The data are supportive of disease models not centered on T cells but centered on the changing nature of the disease over time. PMID- 9228145 TI - Differences in synovial fluid cytokine levels between juvenile and adult rheumatoid arthritis. AB - OBJECTIVE: To evaluate quantitative or qualitative differences in synovial fluid (SF) cytokine levels among patients with systemic juvenile rheumatoid arthritis (JRA), antinuclear antibody positive pauciarticular JRA, or adult RA. METHODS: SF levels of interleukin 1alpha (IL-1alpha), IL-1beta, IL-1 receptor antagonist (IL 1Ra) IL-11, tumor necrosis factor-alpha (TNF-alpha), soluble TNF receptor 1 (sTNFR1), leukemia inhibitory factor (LIF), all measured by immunoassays, and of IL-6, measured with a bioassay using B9 cells, were evaluated in 11 patients with systemic JRA, 24 with pauciarticular JRA, and 22 adult patients with RA. RESULTS: SF IL-6 levels were significantly higher in patients with systemic JRA than patients with pauciarticular JRA (p = 0.003) or RA (p = 0.002). IL-1alpha was detectable in 12/24 SF samples from pauciarticular JRA, in 2/22 SF from RA, and in no sample from systemic JRA (p = 0.004 vs RA; p = 0.005 vs systemic JRA). SF IL-11 levels were significantly higher in patients with RA than patients with systemic JRA (p = 0.012) or pauciarticular JRA (p = 0.005). We found no significant differences in SF levels of IL-1beta, IL-1Ra, TNF-alpha, sTNFR1, or LIF. CONCLUSION: In systemic JRA, SF levels of IL-6 are significantly higher than in pauciarticular JRA or in RA; IL-1alpha is present in a significant proportion of patients with pauciarticular JRA, but not in those with RA or systemic JRA. PMID- 9228146 TI - Adrenal failure in systemic lupus erythematosus. AB - We describe a 39-year-old woman who developed Addison's disease 2 years after diagnosis of systemic lupus erythematosus (SLE). Examining her initial presentation, there is evidence to suggest that adrenal dysfunction may have commenced at the time of diagnosis of SLE. We suggest the need for considering adrenal dysfunction in patients with SLE with systemic complaints. PMID- 9228147 TI - Polymyositis in a patient with thymoma and T cell lymphocytosis. AB - A patient presenting with agranulocytosis and lymphocytosis (4600/mm3) was found subsequently to have polymyositis (PM) and a thymoma. Immunophenotyping of the circulating lymphocytes revealed mature T cells (CD3, CD5, CD7 positive) with normal proportions of both CD4 (55%) and CD8 (38%) positive cells. PM was confirmed by electromyogram and muscle biopsy. Fewer than a dozen cases of thymoma and T cell lymphocytosis have been reported and ours is the first in which autoimmune manifestations have been noted. The presence of lymphocytosis in a patient with PM should prompt the search for a thymoma. PMID- 9228148 TI - Posterior vertebral body erosion by arachnoid diverticula in cauda equina syndrome: an unusual manifestation of ankylosing spondylitis. AB - Cauda equina syndrome is an uncommon complication of longstanding ankylosing spondylitis. It is associated with dorsal arachnoid diverticula, which may erode the lamina and spinous processes of the bony lumbosacral spine. We describe a patient who developed cauda equina syndrome associated with the unusual finding of erosion of the posterior aspect of 2 vertebral bodies by arachnoid diverticula. This was clearly revealed by magnetic resonance imaging of the spine. PMID- 9228149 TI - Gout presenting as non-union of a patellar fracture. AB - We describe a 34-year-old man in whom non-union of a patellar fracture led to an unrecognized diagnosis of gout. He presented with a mass in the superolateral quadrant of the patella, which had the radiographic appearance of a non-united fracture. In the operating room, a gouty tophus was found. Although patellar tophi are uncommon, gout should be included in the differential diagnosis of non united patellar fractures. PMID- 9228150 TI - The Dunlop-Dottridge Lecture: prognosis in inflammatory arthritis: the value of HLA genotyping and the oncological analogy. PMID- 9228151 TI - The future of rheumatology: new directions. PMID- 9228152 TI - Association of NSAID with red cell aplasia. PMID- 9228153 TI - Association of methotrexate, rheumatoid arthritis, and melanoma in 2 patients. PMID- 9228154 TI - Rheumatoid arthritis: failure to isolate or detect mycoplasmas. PMID- 9228155 TI - Myelodysplasia presenting as monoarthritis. PMID- 9228156 TI - Persistence of CD4+ T cells in the arthritic joint after CAMPATH-1H treatment. PMID- 9228157 TI - Arthritis and HIV infection in Kigali, Rwanda, and Paris, France. PMID- 9228158 TI - Systemic lupus erythematosus in childhood. Concordance in a pair of monozygotic twins for anti-dsDNA antibodies and discordance for disease expression. PMID- 9228159 TI - High association between hyperprolactinemia and anticardiolipin antibodies. PMID- 9228160 TI - Sulfamethoxazole/trimethoprim therapy for polymyalgia rheumatica. Report of 5 cases. PMID- 9228161 TI - Association between osteoarthritis of the hand and hip in a skeletal population from London, UK. PMID- 9228163 TI - Scleroderma spectrum disorders, not systemic sclerosis spectrum. PMID- 9228162 TI - The arthritis of Frederic E. Church. PMID- 9228164 TI - Radiation risk--a Chinese perspective: 1996 G. William Morgan Lecture. AB - The great honor of being selected by the Presidents Emeritus Committee of the Health Physics Society as the G. William Morgan Lecturer for 1996 provides me this opportunity to present my lecture at the Plenary Session of the 1996 Health Physics Society Annual Meeting. Under the guidance of Mr. Li Deping, in the late fifties and the early sixties when I began my scientific career, I made a comprehensive literature survey on radiation protection to be aware of this field. Based on this study and on the actual situation in China, I presented the paper "Outline to Health Physics" (Pan 1963), one of the papers I completed in the early times. This gave me a preliminary understanding of the important thoughts of ICRP, NCRP, ICRU and the pioneers of health physics with respect to radiation protection. Preliminary assessment of population dose in China has been done. The natural background radiation sources contribute 93.4% to the total dose, of which is due to radon and thoron progeny. Among the medical exposure from man-made activity, the maximum is from medical exposure (4.21%), the next is from occupational exposure (1.25%), the public exposure contributes about 1.1%. Several practices need to pay special attention: (1) medical exposures, 60.4% of the total increase exposure from man-made activity, (2) the exposure of underground mines, (3) the exposure from coal energy chain, with the sum approximately 100%. The collective dose from nuclear industry contribute 5 x 10( 4). In the coming decade, the doses to the population of China will not significantly increase and may even decrease. PMID- 9228165 TI - Electromagnetic field cancer scares. AB - The internal electric current or power levels that a human is likely to encounter in a typical electromagnetic field environment are calculated. At 60 Hz, a reasonable value for maximum permissible exposure is an internal current density of 1 microA cm(-2). At this frequency, a 600 V m(-1) electric field results in a current density of 0.0002 microA cm(-2), while a 200 microT magnetic field results in 0.6 microA cm(-2). This should be compared with an action potential, which is associated with a current density of 800 microA cm(-2). The ANSI/IEEE C95.1-1991 standards, whose frequency range is 3 kHz to 300 GHz, are examined. Epidemiological studies are briefly considered. The conclusion is that it is highly unlikely that there is a link between electromagnetic fields and cancer. PMID- 9228166 TI - Lost life expectancy rate: an application to environmental levels of radiation. AB - The risk index Lost Life Expectancy Rate (LLER) provides a unitless number (time of life expectancy lost per time exposed) describing the risk from exposure to a given hazard or from partaking in a given activity. Simple equations to calculate the LLER from radiation-induced cancers caused by an exposure to low-level radiation were derived using the relative risk models developed by the Nuclear Regulatory Commission (upper bound estimate), the United Nations Scientific Committee on the Effects of Atomic Radiation-1988, and the National Academy of Science's Committee on the Biological Effects of Ionizing Radiation (BEIR V). Estimates of the LLER to an average person from a continuous exposure to 0.1 microSv h(-1) based on these models, respectively, are 5.5 x 10(-4), 9.5 x 10( 4), and 9.4 x 10(-4). These values compare to LLERs of 0.015 from occupational accidents, 0.25 from being an automobile passenger, and 2.0 from cigarette smoking. Factors effecting LLER from radiation exposures examined in this work include dose rate, age, sex, race and smoking status. PMID- 9228167 TI - A biokinetic model for 137Cs. AB - An improved biokinetic model for 137Cs in humans was developed based on an analysis of data obtained from individuals internally contaminated during an accident in Goiania, Brazil, and other data. Seventeen children (ten girls and seven boys 1-10 y old), ten adolescents (four females and six males), and thirty adults, (fifteen females and fifteen males contaminated in the accident in Goiania contributed to this study. 137Cs retention was determined through periodic measurements in a whole-body counter. In addition to the data on 137Cs retention from these individuals, data from a study on the metabolism of 137Cs in immature, adult, and aged Beagle dogs and data from the literature were used in the formulation of the 137Cs biokinetic model presented. Mathematically, the retention of cesium is described by three exponential terms, and the retention model is based on a step function of body weight. When the ICRP Publication 56 model for cesium was compared to the model suggested in this paper, it was determined that the ICRP model predicts lower effective doses in 5-y-old children and higher effective doses in infants, adolescents, and adults. PMID- 9228168 TI - Behavior of Na131I and meta(131I) iodobenzylguanidine (MIBG) in municipal sewerage. AB - Behavior of 131I activity in primary sludge at the Ann Arbor, Michigan, Municipal Waste Water Treatment Plant was studied in relation to known radioiodine therapy events at the University of Michigan Hospital complex. The principal compounds administered are Na131I, which has widespread use, and meta (131I) iodobenzylguanidine (MIBG), which is a compound unique to the University of Michigan, although labeled antibodies and other forms are also used in therapy and research. The objectives of the study were to determine the environmental fate of such discharges and to determine radiation exposures to workers and the public when sludges are incinerated. Approximately 17% of the MIBG activity administered in a therapy was found in the primary sludge, whereas only 1.1% of the Na131I was in sludge. When land applied, the short half life of 131I in the sludge presents few radiological health concerns; however, incineration, which is done in winter months, is assumed to release organically bound 131I to the atmosphere. Radiation doses due to incineration of sludge containing measured concentrations were calculated for a maximally exposed worker to be 1.7 microSv (0.17 mrem) of which 0.48 microSv (0.048 mrem) was due to a 2-d upset condition. For a more typically exposed worker, and a member of the public, the committed effective dose equivalents were 1.2 microSv (0.12 mrem) and 0.06 microSv (0.006 mrem), respectively, for a 22-wk incineration period with release of all radioiodine in the sludge. Transport time to the treatment plant for radioiodine was found to be much longer than that of normal sewage, possibly due to organic material in sewer lines that absorb iodine. The residence time of radioiodine in the sewer also appears to be longer than expected; whether other radioactive materials are held up the same way is not known but chemical form is surely a factor. PMID- 9228169 TI - A three-dimensional spatial model of plutonium in soil near Rocky Flats, Colorado. AB - The horizontal and depth distribution of plutonium was measured in soil east of the Rocky Flats Environmental Technology Site (formerly the Rocky Flats Plant) near Denver, Colorado, during 1992-1994. The study area was centered on the eastern plume of plutonium contamination and included transects extending from 0.2 km east of the primary origin of the contamination (the 903 Pad) to distances of up to 19 km northeast, east, southeast and south-southeast of the 903 Pad. Soil was collected in 3 cm layers down to 21 cm at exponentially increasing distances along the four transects. Plutonium concentrations decreased rapidly with depth, distance from the 903 Pad, and angle from due east. Depth distributions were independent of distance and angle from the 903 Pad, and our profile model can be used to adjust to a common basis, historical measurements made from sampling to different depths. Based on a total of approximately 1,400 independent measurements, mathematical functions were developed to describe the distance, directional, and depth relationships. These equations, combined with soil density and rock measurements, provided a new method to estimate the plutonium concentration or total deposition per unit area anywhere within the study area. Total deposition per unit area measurements at 50 sites provided an independent test of the model's predictive accuracy. Sampling coefficients of variation based on replicate samples at the main sampling locations averaged 33%, but ranged from 12 to 98%. The analytical measurement coefficient of variation averaged 8%. Mean 0-3 cm soil concentrations of (239,240)Pu among 10 Front Range "background" and 11 community locations near Rocky Flats were 2.1 and 2.3 Bq kg( 1), respectively. PMID- 9228170 TI - Comparison of in situ and laboratory gamma spectroscopy of natural radionuclides in desert soil. AB - In situ and laboratory gamma spectroscopy were used to characterize natural background levels of radiation in the soil at eight sites around the Yucca Mountain Range. The purpose of this practical field analysis was to determine if published empirical in situ calibration factors would yield accurate quantitative specific activities (Bq kg(-1)) in a desert environment. Corrections were made to the in situ calibration factors to account for the on-axis response of a detector with a thin beryllium end window. The in situ gamma spectroscopy results were compared to laboratory gamma spectroscopy of soil samples gathered from each site. Five natural radionuclides were considered: 40K, 214Pb, 214Bi, 208Tl, and 228Ac. The in situ determined specific activities were consistently within +/-15% of the laboratory soil sample results. A quantitative discussion of the factors contributing to the uncertainty in the in situ and laboratory results is included. Analysis on the specific activity data using statistical hypothesis tests determined that three nuclides, 214Pb, 214Bi, and 228Ac showed a weak site dependence while the other two nuclides, 40K and 208Tl, did not exhibit a site dependence. Differing radiation background levels from site to site along with in situ and laboratory uncertainties in excess of 10% are two factors that account for the weak site dependence. Despite the good correlation between data, it was recommended that the in situ detector be calibrated by a detector-specific Monte Carlo code which would accurately model more complex geometries and source distributions. PMID- 9228171 TI - A trace metal (zinc and iron) study on low dose x-radiation response in rat skin. AB - There is no reliable bio-dosimeter regarding low dose radiation effects in mammalian systems. In this study, chronic low dose (< 1 cGy) whole body x irradiated rat skin have shown altered trace metal (zinc and iron) content which clearly indicated the redistribution of these metals in the integumentary system. The decreased zinc to iron ratios suggested enhanced oxidative stress of the tissue. Changes in trace metal content in irradiated rat skin, as a biological response to low dose radiation, were non-linear. Moreover, the lowered zinc content of E2, E3, E4 and E5 dose groups suggested a different steady state, compared to the control. The Zn: Fe ratio decreased with increasing radiation dose. PMID- 9228172 TI - Radioactivity in zircon and building tiles. AB - Zircon (ZrSiO4) is commonly used in the manufacture of glazed tiles. In this study we found high concentrations of the radionuclides 226Ra, 232Th, 40K in zircon sand. The average radium equivalent (A(Ra) + 1.26 A(Th) + 0.086 A(k)) in zircon sand is 17,500 Bq kg(-1), which is 106 times as much as that in ordinary building materials. The external radiation (gamma + beta) dose rates in air at 5 cm from the surface of piles of zircon sand sacks range from 1.1 to 4.9 x 10(-2) mGy h(-1) with an average of 2.1 x 10(-2) mGy h(-1). Although no elevated gamma ray radiation or radon exhalation rate was detected in rooms decorated with glazed tiles, which is characteristic of combined alpha, beta and gamma emitting thin materials, the average gamma-ray radiation dose rate at the surface of the tile stacks in shops is 1.5 times as much as the indoor background level. The average area density of total beta emitting radionuclides in glazed floor tiles and glazed wall tiles is 0.30 Bq cm(-2) and 0.28 Bq cm(-2), respectively. It was estimated that the average beta dose rates in tissue at a depth 7 mg cm(-2) with a distance 20-100 cm from the floor tiles were 3.2 to 0.9 x 10(-7) Gy h(-1). The study indicates that the beta-rays from glazed tiles might be one of the main factors leading to an increase in ionizing radiation received by the general public. Workers in glazed tile manufacturing factories and in tile shops or stores may be exposed to elevated levels of both beta-rays and gamma-rays from zircon sand or glazed tile stacks. No elevated radiation from unglazed tiles was detected. PMID- 9228173 TI - A survey of radon and thoron progeny for dwellings in Hong Kong. AB - The potential alpha energy concentrations of radon and thoron progeny have been surveyed for dwellings in Hong Kong and the mean values are obtained as 3.58 and 2.29 mWL, respectively. The relative importance of the value for thoron is unexpectedly high, which is attributed to the high 232Th content of the building materials used in Hong Kong. It has also been found that the potential alpha energy concentration values for radon progeny changed dramatically with the season due to the different aerosol contents in the air in different seasons. The factors affecting the potential alpha energy concentration values have also been studied. These factors fall into three categories, namely (1) the building characteristics including age of the buildings, wall coverings and floor coverings; (2) the location of sites including nearby environments and the elevation of the sites; and (3) the meteorological parameters including wind speed, atmospheric pressure, air temperature and relative humidity. For categories (1) and (2), all factors seem to affect the potential alpha energy concentration values, although the effects may be different for radon and thoron progeny, which may be due to the very much different half lives of radon and thoron gas and to the different behavior of radon and thoron progeny in the attachment to aerosols. For category (3), only wind speed has been found to have effects. PMID- 9228174 TI - Ethical issues in radiation protection. AB - In this note the authors survey existing international radiation-protection recommendations of the ICRP, the IAEA, and the ILO. After outlining previous work on the ethics of radiation protection and risk assessment/management, the authors review ethical thinking on five key issues related to radiation protection and ethics. They formulate each of these five issues in terms of alternative ethical stances: (1) Equity vs. Efficiency, (2) Health vs. Economics, (3) Individual Rights vs. Societal Benefits, (4) Due Process vs. Necessary Sacrifice, and (5) Stakeholder Consent vs. Management Decisions. PMID- 9228175 TI - Safety systems in gamma irradiation facilities. AB - A new electronic device has been developed to guard against individuals gaining entry through the product entry and exit ports into our irradiation facility for industrial sterilization. This device uses the output from electronic sensors and pressure mats to assure that only the transport cabins may pass through these ports. Any intention of personnel trespassing is detected, the process is stopped by the safety system, and the source is placed in safe position. Owing to a simple construction, the new device enables reliable operation, is inexpensive, easy to implement, and improves the existing safety systems. PMID- 9228176 TI - Radioactivity in sediments of the Karnaphuli river estuary and the Bay of Bengal. AB - Sediment samples from the Karnaphuli river estuary, nearshore, and off-shore regions off the coast of Chittagong in the Bay of Bengal were analyzed for the natural radioactivity contents of 232Th, 238U and 40K and anthropogenic radioactivity contents of 137Cs and 134Cs using HPGe gamma spectrometry, together with the measurement of sediment pH and grain size analyses of the collected samples. The activity of 232Th found in sediment ranged from 10.44 +/- 2.31 to 64.02 +/- 8.13 Bq kg(-1), 238U activity ranged from 5.87 +/- 1.21 to 27.85 +/- 1.71 Bq kg(-1), 40K activity from 118.28 +/- 19.70 to 608.21 +/- 75.70 Bq kg(-1), and the activity 137Cs ranged from 0.09 +/- 0.06 to 4.64 +/- 0.19 Bq kg(-1), no 134Cs radioactivity was detected at any of the sampling stations. PMID- 9228177 TI - Energy deposition pattern from tritium and different energy photons--a comparative study. AB - Energy deposition patterns of beta particles from tritium decay have been analyzed in comparison with photon radiation in the energy range from 12 to 1250 keV. Energy deposition was modeled by Monte Carlo means using electron tracks in water vapor with a complete follow up of delta electrons. Ionizations have been used as representative of track structure. Frequencies of clusters on a nanometer scale and their spatial distribution in spherical targets representing cell nuclei have been derived. Cluster analysis has been carried out by implementing a k-means method. Frequencies of ionization clusters from tritium are similar to 60 100 keV photons. Spatial distribution of clusters from tritium closely match approximately 70 keV photons up to 2 microm separation. PMID- 9228178 TI - Residential radon and lung cancer in Sweden. PMID- 9228179 TI - Residential radon and lung cancer in Sweden. PMID- 9228181 TI - We appear to have done it to ourselves again. PMID- 9228180 TI - Tritium exposure from radioluminous watches. PMID- 9228182 TI - Relative biological effectiveness for organically bound tritium. PMID- 9228183 TI - Purinoceptor-stimulated phosphoinositide hydrolysis in Madin-Darby canine kidney (MDCK) cells. AB - Extracellular nucleotides, acting through P2-purinoceptors, have been implicated in the regulation of ion transport in epithelia, including Madin-Darby canine kidney (MDCK) cells. In this study, experiments were conducted to characterize the P2-purinoceptor subtype on MDCK cells responsible for stimulating inositol phosphate (IP) accumulation using a range of nucleotide analogues. In Ca2+- and Mg2+-free Krebs-Henseleit solution (KHS), ATP, UTP, and ATPgammaS caused an increase in IP accumulation as a function of concentration with comparable kinetics. The order of potency for the nucleotide analogues was UTP = ATPgammaS > ATP = 2-chloro ATP (Cl-ATP) >> alpha,beta-methylene ATP (alpha,beta-MeATP) = 2 methylthio ATP (2MeSATP). Selective agonists for P1-, P2X- and P2Y-purinoceptors, such as N6-cyclopentyl adenosine, AMP, alpha,beta-MeATP, and 2MeSATP, had little effect. Stimulation of MDCK cells with maximally effective concentrations of ATP and UTP showed no additive effect and furthermore, ATP, UTP, and ATPgammaS induced cross-desensitization of the IP response, suggesting that ATP and UTP act upon a common nucleotide receptor, i.e. a P2U-purinoceptor. In Ca2+- and Mg2+ containing KHS, the concentration-response curves of ATP, UTP, and ATPgammaS were shifted to the right of those obtained in Ca2+- and Mg2+-free buffer, and asymptotic maxima were not reached, indicating that ATP4- and not MgATP2- or CaATP2- was the active agonist. Pretreatment of MDCK cells with pertussis toxin (PTX) inhibited ATP- and UTP-induced IP accumulation in a concentration-dependent fashion but did not completely abolish the IP accumulation, indicating that a PTX sensitive G protein was partially involved in the IP response. In conclusion, ATP and UTP-stimulated IP accumulation in MDCK cells appears to be mediated through the activation of P2U-purinoceptors coupled to a G protein that is partially sensitive to PTX. A form of nucleotide uncomplexed with divalent ions such as ATP4- seems to be the preferential agonist form for the purinoceptors on MDCK cells. PMID- 9228184 TI - Inhibitors of ser/thr phosphatases 1 and 2A induce apoptosis in pituitary GH3 cells. AB - Two structurally different inhibitors of ser/thr phosphatases 1 and 2A, okadaic acid and calyculin A, time- and concentration-dependently stimulated and inhibited cell-specific function (hormone gene expression) in pituitary GH3 cells. The negative effect was associated with the appearance of apoptotic cell death. Nanomolar concentrations of both agents produced the characteristic morphological alterations and a DNA fragmentation ladder. Calyculin A treatment resulted in comparable changes with 10fold lower concentrations than okadaic acid. Observations with derivatives of okadaic acid with no or lower phosphatase inhibitory potency supported the conclusion that apoptosis induction is related to inhibition of ser/thr phosphatases, presumably types 1 and 2A. Membrane damage as measured by lactate dehydrogenase liberation into medium was significantly lower in apoptotic vs. necrotic cells. DNA fragmentation could be reduced by the addition of zinc but not by removal of extracellular calcium with EGTA. Apoptotic changes were reduced by the concomitant activation of protein kinase A by a membrane permeable cAMP analogue. Incubation of cells for 4 months in successively increased concentrations of okadaic acid resulted in a population that proliferated at the initially lethal concentration of 30 nM. PMID- 9228186 TI - Effects of chronic treatment with fluoxetine and citalopram on 5-HT uptake, 5 HT1B autoreceptors, 5-HT3 and 5-HT4 receptors in rats. AB - The effect in rats of chronic treatment with two specific 5-HT reuptake inhibitors (SSRI) with antidepressant properties, citalopram (10 mg/kg, i.p. twice a day for 14 days, one day washout) and fluoxetine (15 mg/kg, p.o. twice a day for 21 days, 7 days washout), was evaluated on some mechanisms involved in central 5-HT neurotransmission. No adaptive modifications of brain 5-HT uptake (sites) were found by measuring functional [3H]5-HT uptake and [3H]citalopram binding in cortical and hippocampal synaptosomes, and by [3H]citalopram binding autoradiography in the raphe nuclei (5-HT cell bodies) and the ventral tegmental area (5-HT axonal pathway). Chronic treatments had no effect on presynaptic 5 HT1B autoreceptors, functionally evaluated by measuring 5-HT1B-mediated inhibition of depolarization-induced [3H]5-HT release from cortical and hippocampal synaptosomes. Chronic citalopram or fluoxetine did not significantly affect the binding of [3H]BRL-43694 to 5-HT3 receptors in the rat brain cortex. Citalopram had no effect on [125I]SB-207710 binding to 5-HT4 receptors, measured by autoradiography in the substantia nigra. Negative results, such as those reported in the present study, could be due to a number of variables including the animal species, the treatment schedule or the brain areas considered, thus explaining the differences from some previous reports of significant effects of SSRI. However, our negative data are in agreement with many other published studies, suggesting that adaptive modifications of brain 5-HT transporters, terminal 5-HT1B receptors, 5-HT3 and 5-HT4 receptors may not be a general effect induced by all SSRI. PMID- 9228185 TI - Multiple [3H]-nemonapride binding sites in calf brain. AB - [3H]-Nemonapride has been the ligand of choice to label D4 dopamine receptors. Its specificity was questioned when it was discovered that sigma (sigma) sites were also labeled by [3H]-nemonapride. To further characterize the binding of [3H]-nemonapride, three areas of calf brain (striatum, frontal cortex and cerebellum) were examined. In all three areas, [3H]-nemonapride labeled multiple sites. Dopaminergic and sigma sites were the most prominent. The sigma binding profile was sigma-1 like with a Ki binding profile as follows (in order of decreasing potency): haloperidol, PPAP, pentazocine, DTG, U-50488, R(+)-3-PPP. Experiments using sulpiride and pentazocine to block striatal dopaminergic and sigma sites, respectively, revealed additional, not previously characterized binding sites for [3H]-nemonapride. One component which was present in striatum but not in frontal cortex or cerebellum, had affinity for some neuroleptics and WB-4101, but not for typical serotonergic agents. Thus, [3H]-nemonapride has no selectivity for dopamine receptors unless stringent experimental conditions are met. PMID- 9228187 TI - Differential desensitization of ionotropic non-NMDA receptors having distinct neuronal location and function. AB - The release of tritium from rat hippocampal synaptosomes prelabeled with [3H]noradrenaline ([3H]NA) or [3H]5-hydroxytryptamine ([3H]5-HT) and from rat neocortex synaptosomes prelabeled with [3H]choline and the release of endogenous GABA and glutamate from rat neocortex synaptosomes were monitored during superfusion with media containing varying concentrations of alpha-amino-3-hydroxy 5-methyl-4-isoxazolepropionic acid (AMPA) or kainic acid. Concentration-dependent release potentiations were elicited by both excitatory amino acids (EAAs) in all the transmitter systems investigated. The releases evoked by 100 microM AMPA were, in all cases, almost totally dependent on external Ca2+ and sensitive to 6.7-dinitroquinoxaline-2,3-dione (DNQX), indicating involvement of non-NMDA receptors. When cyclothiazide, a drug able to prevent desensitization of AMPA preferring receptors, was added to the superfusion medium (at 1 or 10 microM) concomitantly with 100 microM AMPA or kainate, the EAA-evoked release of [3H]NA was significantly enhanced. Concanavalin A, a lectin thought to prevent desensitization of kainate-preferring receptors, had no effect (up to 10 microM) on the release of [3H]NA evoked by AMPA or kainate. The effect of cyclothiazide was lost if, after an 8-min pretreatment, the drug was removed just before the AMPA stimulus. When added concomitantly with the EAAs, cyclothiazide potentiated the release of [3H]5-HT elicited by AMPA and, less so, that evoked by kainate. Concanavalin A was ineffective. Neither cyclothiazide (1 or 10 microM) nor concanavalin A (3 or 10 microM) could affect the release of [3H]ACh or endogenous GABA provoked by 100 microM AMPA or kainate, suggesting that the receptors involved do not desensitize. Exposure of neocortex synaptosomes to AMPA or kainate concomitantly with cyclothiazide caused endogenous glutamate release that did not differ from that evoked by the EAAs alone. In contrast, the effects of AMPA and kainate were potentiated by concanavalin A. The activity of the lectin (3 microM) persisted when it was applied for 8 min and then removed before the AMPA or kainate (100 microM) pulse. When hippocampal synaptosomes prelabeled with [3H]NA were subjected to three subsequent AMPA (100 microM) stimuli (S1, S2 and S3), the release of [3H]NA decreased dramatically from S1 to S3 (S3/S1 = 0.14 +/- 0.04); a significant 'protection' of the AMPA effect was offered by 1 microM cyclothiazide (S3/S1 = 0.36 +/- 0.06). This value did not differ from the S3/S1 ratio (0.38 +/- 0.04) obtained in parallel experiments with 12 mM K+. The release evoked by high-K+ was insensitive to cyclothiazide. Finally, the effect of AMPA on the release of [3H]ACh did not respond to cyclothiazide also during three subsequent stimuli with 100 microM AMPA. To conclude: a) ionotropic non-NMDA receptors mediating enhancement of NA, 5-HT, ACh, GABA and glutamate release exist on the corresponding nerve terminals; b) the receptors present on noradrenergic and serotonergic neurons are AMPA-preferring receptors, whereas the glutamate autoreceptors resemble most the kainate-preferring subtype; the receptors mediating ACh and GABA release can not be subclassified at present; c) desensitization may not be a property of all non-NMDA ionotropic receptors. The receptors here characterized represent five models of native non-NMDA receptors suitable for pharmacological and molecular studies. PMID- 9228188 TI - Labelling of I2B-imidazoline receptors by [3H]2-(2-benzofuranyl)-2-imidazoline (2 BFI) in rat brain and liver: characterization, regulation and relation to monoamine oxidase enzymes. AB - The novel selective imidazoline radioligand [3H]2-(2-benzofuranyl)-2-imidazoline (2-BFI) was used to characterize and assess further the nature of I2-imidazoline receptors in rat brain and liver. In the cerebral cortex, 2-BFI displayed high affinity (Ki = 9.8 nM) for a single class of [3H]2-BFI binding sites. Other imidazoline/guanidine compounds (e.g. aganodine, cirazoline and idazoxan) displayed biphasic competition curves, indicating the existence of high (KiH = 2.9-78 nM; R(H) = 61-83%) and low (KiL = 4.7-158 microM) affinity sites. The pharmacological profile for [3H]2-BFI binding (aganodine > cirazoline > 2-BFI >> clonidine > amiloride >> efaroxan) was typical of that for I2-sites. This profile was almost identical to that obtained against [3H]idazoxan (correlation between pKi values, r = 0.97) which indicated that the sites characterized with [3H]2-BFI in brain corresponded to I2-imidazoline receptors. The low affinity of amiloride against [3H]2-BFI (Ki = 900 nM) further indicated that these brain I2-sites belong to the I2B-subtype. [3H]2-BFI binding sites (Bmax = 72 fmol/mg protein) in brain were differentially modulated by treatment (7 days) with cirazoline (up regulation: 25%) and the MAO inhibitor phenelzine (down-regulation: 31%), indicating that these I2-sites are regulated in vivo, as is the case for those labelled by [3H]idazoxan. Chronic treatment with 2-phenylethylamine, a phenelzine metabolite and endogenous amine, did not alter the density of brain of I2 imidazoline receptors labelled by [3H]idazoxan. Preincubation of liver membranes with the MAO inhibitor clorgyline (10(-7) M) abolished the binding of [3H]Ro 41 1049 (N-(2-aminoethyl)-5-(m-fluorophenyl)-4-thiazole carboxamide) to MAO-A, but it did not alter the binding of [3H]Ro 19-6327 (N-(2-aminoethyl)-5-chloro-2 pyridine carboxamide) to MAO-B or that of [3H]2-BFI to I2-sites. At 10(-4) M it also abolished MAO-B sites, but a substantial proportion of I2-sites (40%) remained intact. Preincubation of liver membranes at 60 degrees C also abolished MAO-A/B sites, whereas still 22% of I2-sites remained. The results indicate that [3H]2-BFI is a good tool for the identification of I2-imidazoline receptors and suggest further that certain I2-sites and MAO are different proteins. PMID- 9228190 TI - Vigilance and EEG power in rats: effects of potent inhibitors of the neuronal nitric oxide synthase. AB - We examined the effects of potent neuronal nitric oxide synthase inhibitors, 3 bromo-7-nitro indazole (3-Br-7-NI) and S-methyl-L-thiocitrulline (S-Me-TC) on general behaviour, vigilance stages and electroencephalographic (EEG) power spectra in rats. In addition, we studied the effect of 7-nitro indazole (7-NI) on EEG power spectra in rats during dark and light periods. 3-Br-7-NI induced ptosis and decrease of slow wave sleep and rapid eye movement sleep in the rat. 7-NI and 3-Br-7-NI reduced the EEG power density in all frequency bands in the rat, suggesting a depression of central neuronal activity. This effect of 7-NI was more prominent during the day than during the night, indicating a circadian variation in the nitric oxide synthase (NOS) response to NOS inhibitor. EEG power was the most reduced in the 7-9 Hz range of the rhythmic slow activity (theta rhythm), which is in accordance with decreased locomotion observed following administration of NOS inhibitors. Although S-Me-TC is the most potent NOS inhibitor in vitro experiments, it had less effect on vigilance and EEG power in the rat than other NOS inhibitors used in this study, probably due to its short lasting and blood pressure raising effect. The present results indicate that nitric oxide exerts an excitatory and circadian dependent effect in the central neuronal structures involved in the regulation of vigilance. PMID- 9228189 TI - Convulsant effects of some xanthine derivatives in genetically epilepsy-prone rats. AB - The behavioural and electrocorticographic (ECoG) convulsant effects of several xanthine derivatives injected intraperitoneally (i.p.) were studied in genetically-epilepsy prone rats. The aim of the study was to evaluate the relationship among convulsant potency, molecular structure and lipophilicity of some xanthines. Animals were injected i.p. with various doses (250-1000 micromol/kg) and a different convulsant potency was observed among the various xanthines tested. IBMX (3-isobutyl-1-methylxanthine), theophylline (1,3 dimethylxanthine) and caffeine (1,3,7-trimethylxanthine) induced an epileptogenic pattern that consisted in an initial phase characterized by wet-dog shakes followed by head tremor, nodding, clonic convulsion and they appeared to be the most potent xanthines among those studied. During seizures, the electrocortical activity was usually characterized by single or multiple sharp- or spike-wave episodes followed by polyspike discharges. After the highest doses of IBMX, theophylline and caffeine, the animals react with falling down, transient tonic clonic seizures, escape response and generalized seizures followed by post-ictal period. Equimolar doses of 8-chlorotheophylline and theobromine (3,7 dimethylxanthine) produced less evident epileptic responses in comparison to previous compounds, whereas no epileptic signs were observed following the administration of enprofylline (3-propylxanthine), etofylline [7-(2 hydroxyethyl)theophylline], diprophylline [7-(2,3-dihydroxy-propyl)theophylline] and doxofylline [7-(1,3-dioxolan-2-ylmethyl) theophylline]. Lipophylicity of the compounds was determined, but no convincing correlations were found between the rank order of lipophilicities and the convulsant potencies of the compounds studied. On the other hand, structure-activity relationship was also investigated. We suggest that the substitution pattern on the xanthine nucleus may explain, in part, the different convulsant potency of the compounds studied. Furthermore, a selective antagonism of adenosine subtype receptors should be considered. PMID- 9228191 TI - Influence of alpha- and beta-adrenoceptor antagonists on ventricular fibrillation in ischemic rat hearts. AB - Although it is generally accepted that adrenergic influences contribute to arrhythmias during myocardial ischemia, it is still a matter of debate whether these arrhythmogenic effects are mediated via alpha- or beta-adrenergic receptors and which particular receptor subtype is involved. Controversial results may be due to ancillary properties of the adrenergic receptor antagonists used to resolve this question. Therefore, we compared the influence of various, structurally different alpha- and beta-adrenoceptor blocking agents on the occurrence of ventricular fibrillation in rat isolated hearts. Regional ischemia was induced by ligature of the left coronary artery and ECG was monitored over 30 min of coronary occlusion. Myocardial ischemia precipitated ventricular fibrillation in a well reproducible manner with an incidence of about 80% in control hearts. Racemic propranolol (0.1-1 micromol/l) concentration-relatedly reduced the incidence of ventricular fibrillation from 71% in controls to 10%, whereas the beta-adrenoceptor blocking agents atenolol (10 micromol/l) and timolol (1 micromol/l) did not influence the occurrence of arrhythmias. Moreover, both stereoisomers of propranolol were equipotent in suppressing ischemia-induced ventricular fibrillation, indicating an action of propranolol independent of its beta-adrenoceptor blocking properties. Unselective antagonism of alpha adrenoceptors by phentolamine decreased the incidence of ventricular fibrillation from 90% to 58% at 0.1 micromol/l and totally suppressed ventricular fibrillation at 1 micromol/l. The alpha1-selective antagonists prazosin and HEAT concentration dependently (0.1-10 micromol/l) reduced the incidence of ventricular fibrillation from 83% to 0%, whereas the alpha2-selective antagonist RX 821002 revealed no antiarrhythmic effect. Furthermore, subtype specific antagonism of alpha1A adrenoceptors by WB 4101 clearly reduced the occurrence of ventricular fibrillation in a concentration-dependent manner (0.01-1 micromol/l) from 66% to 17%. Conversely, CEC, known to block the alpha1B-adrenoceptor subtype, possessed no antifibrillatory effect. In conclusion, the contribution of catecholamines to ischemia-induced arrhythmias in rat isolated heart is primarily mediated via WB 4101-sensitive alpha1-adrenergic activation. Beta- and alpha2-adrenoceptor blockade did not affect ventricular fibrillation in this model. The antifibrillatory action of propranolol was rather due to ancillary properties of this agent. PMID- 9228192 TI - Effects of adenosine and adenosine analogues on mean circulatory filling pressure and cardiac output in anesthetized rats. AB - The effects of adenosine and adenosine analogues, 2-[p-(2-[carboxyethyl) phenethylamino]-5'-N-ethylcarboxamidoadenosine (CGS 21680) and N6-2-(4 aminophenyl)-ethyladenosine (APNEA), on mean arterial pressure, cardiac output, mean circulatory filling pressure, arterial resistance, venous resistance and heart rate in untreated or treated (with ganglion-blockers mecamylamine and atropine) pentobarbital-anesthetized rats were examined. Infusion of adenosine (100, 300 and 900 microg/kg/min), CGS 21680 (0.1, 0.3 and 0.9 microg/kg/min) or APNEA (1.0, 3.0 and 9.0 microg/kg/min) reduced mean arterial pressure and arterial resistance in all groups. Adenosine and APNEA also reduced mean circulatory filling pressure, venous resistance and heart rate in untreated animals. Furthermore, APNEA but not adenosine reduced cardiac output. In contrast, CGS 21680 increased cardiac output and heart rate but did not have any effect on mean circulatory filling pressure or venous resistance. In ganglion blocked rats, APNEA reduced cardiac output, mean circulatory filling pressure and heart rate, while adenosine did not have any effect on these parameters. In addition, APNEA and adenosine reduced arterial resistance but were unable to alter venous resistance while CGS 21680 reduced mean circulatory filling pressure and arterial resistance but did not further affect cardiac output, heart rate and venous resistance in ganglion-blocked animals. The results of the present study suggest that adenosine and APNEA dilate arterioles and vein, whereas CGS 21680 causes arterial dilatation but not venodilatation in untreated animals due to hypotension-induced sympathetic activation. A possible explanation for the present observations could be differences in the distribution of vascular A2 versus A3 adenosine receptors in the venous circulation. PMID- 9228193 TI - Actions of the antiarrhythmic peptide AAP10 on intercellular coupling. AB - Disturbances in gap junction distribution and a decrease in the connexin43 content of the heart were shown to occur after myocardial infarction and in ischemic heart disease, respectively. These changes are now thought to play an important role in the genesis of arrhythmias associated with these diseases. It is thought that agents that can increase cellular coupling might be beneficial in these situations. Recently, we presented data showing that the synthetic peptide AAP10 acts antiarrhythmically in a model of regional ischemia. The data suggested that AAP10 might act via an increase in cellular coupling. The goal of this study was to establish whether AAP10 can interact with cardiac gap junctions. Measurements of the stimulus-response-interval (SRI) in guinea pig papillary muscle showed that high concentrations of AAP10 (1 microM) can decrease the SRI by about 10% under normoxic conditions. At lower concentrations (10 nM) AAP10 had no effect on SRI under normoxic conditions but prevented the increase in the SRI induced by perfusion with hypoxic, glucose-free Tyrode's solution. Double-cell voltage-clamp experiments confirmed that AAP10 can interact with cardiac gap junctions. 10 nM AAP10 could either diminish or reverse the run-down of gap junction conductance normally observed in pairs of guinea pig ventricular myocytes. During control gap junction conductance decreased with a rate of -2.5 +/- 2.0 nS/min. After application of 10 nM AAP10 gap junction conductance increased with a rate of +1.0 +/- 0.7 nS/min (p < 0.01). After washout of AAP10 gap junction conductance decreased again with a rate not significantly different from control. Our results show that AAP10 does interact with gap junctions. Because no other effects of AAP10 on other electrophysiological parameters could be found, this action on gap junctions might be the basis of AAP10's antiarrhythmic effect seen in previous studies. PMID- 9228194 TI - Capacitative Ca2+ entry associated with alpha1-adrenoceptors in rat aorta. AB - In rat aorta, depletion of internal Ca2+ stores by addition of noradrenaline (1 microM) induces a biphasic response (an initial phasic response and a tonic one) mediated by two different intracellular Ca2+ pools. This response cannot be repeated, suggesting a depletion of internal Ca2+ stores sensitive to noradrenaline. In absence of the agonist, this depletion is the signal for the entry of extracellular Ca2+, not only to refill the stores but also, under our experimental conditions, to activate the contractile proteins thus inducing an increase in the resting tone (IRT) that constitutes functional evidence of this Ca2+ entry. The ionic channels involved in the mechanism of the IRT have been studied in the present work. The fact that the addition of nimodipine (10(-15) 10(-11) M) selectively inhibits the IRT suggests that this mechanical response is mediated by Ca2+ influx through dihydropyridine-sensitive Ca2+ channels. Moreover, the inhibitory action of nimodipine is attenuated by glibenclamide (10 microM). Cromakalim (10(-10)-10(-6) M) also inhibits the IRT concentration dependently, and this inhibition is antagonized by glibenclamide (10 microM). These results relate the ATP-dependent K+ channels to the mechanism of the IRT. The refilling of the two internal Ca2+ compartments sensitive to noradrenaline was, like the IRT, altered in presence of the compounds tested, since the subsequent contractile response to noradrenaline was decreased. The present results suggest that nimodipine treatment inhibits the refilling of the Ca2+ compartment responsible for the tonic contraction induced by noradrenaline in Ca2+-free medium, whereas the refilling of the Ca2+ pool responsible for the phasic response to noradrenaline remained unaltered. Both the phasic and tonic responses to noradrenaline in Ca2+-free medium decreased after treatment with cromakalim. We can therefore assume that the refilling of both Ca2+ compartments sensitive to noradrenaline was inhibited. In conclusion, these results are consistent with the contraction of the rat aorta in response to noradrenaline in Ca2+-free medium consisting of an initial phasic response and a tonic one. The former is due to the release of internal Ca2+ from a compartment refilled through a special channel that is cromakalim but not dihydropyridine sensitive. The tonic response is due to Ca2+ release from another compartment refilled through a cromakalim- and dihydropyridine-sensitive Ca2+ channel. The Ca2+ entry through this latter channel intervenes in the IRT observed during the refilling of these stores previously depleted by noradrenaline, and the opening state of this channel is also modulated by ATP-dependent K2+ channels. PMID- 9228195 TI - Nature of 5-HT1-like receptors mediating depressor responses in vagosympathectomized rats; close resemblance to the cloned 5-ht7 receptor. AB - It has been suggested that the late hypotensive response to serotonin (5 hydroxytryptamine; 5-HT) in vagosympathectomized rats is mediated by '5-HT1-like' receptors since this effect is mimicked by 5-carboxamidotryptamine (5-CT), is not modified by cyproheptadine, ketanserin or MDL 72222, but it is blocked by methysergide. The present study was set out to reanalyze this suggestion in terms of the classification schemes proposed in 1994 and 1996 by the NC-IUPHAR subcommittee on the classification and nomenclature of 5-HT receptors. I.v. bolus injections of 5-CT (0.01-0.3 microg x kg(-1)), 5-HT (1-30 microg x kg(-1)) and 5 methoxytryptamine (5-MeO-T; 1-30 microg x kg(-1)) produced dose-dependent hypotensive responses with a rank order of agonist potency: 5-CT >> 5-HT > 5 methoxytryptamine with sumatriptan (30-1000 microg x kg(-1)) inactive. The depressor responses to 5-HT and 5-CT were not attenuated by i.v. GR127935 (300 3000 microg x kg(-1)) or equivalent volumes of saline. In contrast, lisuride, methiothepin, mesulergine, metergoline and clozapine dose-dependently antagonized the responses to 5-HT and 5-CT; the rank order of apparent pA2 values against 5 HT and 5-CT, respectively, was: lisuride (7.7; 7.8) > methiothepin (6.8; 7.0) > or = mesulergine (6.4; 6.6) > clozapine (5.7; 5.8); metergoline displayed variable potencies (5.6; 6.4). Except for lisuride, which also affected isoprenaline-induced hypotension, the antagonism by the other drugs was selective. Based upon the above rank order of agonist potency, the blockade by a series of drugs showing high affinity for the cloned 5-ht7 receptor and the lack of blockade by GR127935, our results indicate that the 5-HT receptor mediating hypotension in vagosympathectomized rats is operationally similar to other putative 5-ht7 receptors mediating vascular and non-vascular responses (e.g. relaxation of the rabbit femoral vein, canine coronary and external carotid arteries and guinea-pig ileum as well as feline tachycardia). PMID- 9228196 TI - Influence of atropine on the cardiovascular effects of noradrenaline and tyramine in elder volunteers. AB - The aim of this study, carried out in six elder healthy volunteers (mean age: 61 years), was to determine the influence of muscarinic receptor blockade with atropine (15 microg/kg i.v. loading dose followed by 0.15 microg/kg/min by i.v. infusion) on the effects of i.v. infusions of noradrenaline (5 incremental doses of 10-120 ng/kg/min) or tyramine, that releases endogenous noradrenaline (4 incremental doses of 5-20 microg/kg/min), on blood pressure, heart rate and systolic time intervals (STI's, as a measure of positive inotropism). These results were compared with those recently published for young healthy volunteers (mean age: 26 years; Schafers et al. 1997). Noradrenaline caused increases in systolic and diastolic blood pressure, decreases in heart rate and a shortening of STI's that were not different from those in young volunteers. Atropine did not significantly affect these hemodynamic responses to noradrenaline, while in young volunteers it significantly enhanced noradrenaline-induced blood pressure increases and converted the heart rate decrease into an increase. In the present study in elder volunteers, tyramine caused a smaller increase in systolic blood pressure than in the previous study in young volunteers; in addition, it slightly increased diastolic blood pressure while it decreased diastolic blood pressure in young volunteers. Atropine did not significantly affect the hemodynamic effects of tyramine in the elder volunteers, while in the young volunteers it enhanced the increase in systolic blood pressure and converted the decreases in diastolic blood pressure and heart rate into increases. These results indicate a) that ageing is accompanied by a blunted baroreflex-mediated parasympathetic activation resulting in reduced cholinergic vasodilation and decreases in heart rate, and b) that ageing is associated with a decreased responsiveness of (cardiac) beta adrenoceptors and (vascular) alpha1-adrenoceptors which is only unmasked when the counterregulatory action of parasympathetic activation is removed. PMID- 9228198 TI - 1,1'-Diisopropyl-2,4'-cyanine (disprocynium24), a potent uptake2 blocker, inhibits the renal excretion of catecholamines. AB - 1,1'-Diisopropyl-2,4'-cyanine (disprocynium24), a potent inhibitor of the extraneuronal monoamine transport system (uptake2), was previously shown to reduce the clearance of catecholamines from plasma not only by blocking uptake2 but presumably also by blocking organic cation transport. To provide more direct evidence for the latter conclusion, the present study was carried out in anaesthetized rabbits. It aimed at determining the effect of disprocynium24 on the renal excretion of catecholamines which is known to be, at least in part, a consequence of organic cation transport in the kidney. To this end, the plasma clearance due to renal excretion (Cl(u)) of endogenous as well as infused 3H labelled adrenaline, noradrenaline and dopamine was determined for 60-min periods of urine collection in rabbits treated either with disprocynium24 (270 nmol kg( 1) i.v. followed by i.v. infusion of 80 nmol kg(-1) min(-1)) or vehicle. Two groups of animals were studied: group I (monoamine oxidase and catechol-O methyltransferase intact) and group II (monoamine oxidase and catechol-O methyltransferase inhibited). A third group of animals with intact monoamine oxidase and catechol-O-methyltransferase was used to study the effect of disprocynium24 on the glomerular filtration rate (as determined by measuring the plasma clearance of inulin). In vehicle controls, Cl(u) of endogenous adrenaline, noradrenaline and dopamine was 7.2, 5.2 and 153.6 ml kg(-1) min(-1), respectively, in group I and 10.4, 7.0 and 134.3 ml kg(-1) min(-1), respectively, in group II. Similar control values of Cl(u) were obtained for infused 3H adrenaline and 3H-noradrenaline, but not for infused 3H-dopamine; Cl(u) of 3H dopamine (4.9 ml kg(-1) min(-1) in group I and 15.4 ml kg(-1) min(-1) in group II) was considerably smaller than Cl(u) of endogenous dopamine, indicating that most of the dopamine in urine (i.e., 98% in group I and 92% in group II) was derived from the kidneys rather than from the circulation. By contrast, only about one quarter of the noradrenaline in urine (32% in group I and 24% in group II) and none of the urinary adrenaline were of renal origin. In both groups, disprocynium24 markedly reduced the Cl(u) of endogenous catecholamines (by 72 90%) and of infused 3H-catecholamines (by 49-69%). Moreover, it preferentially inhibited the renal excretion of those components of urinary dopamine and noradrenaline which were derived from the kidney. Therefore, disprocynium24 inhibits the tubular secretion of catecholamines and, hence, organic cation transport in the kidney. This conclusion was substantiated by the observation that disprocynium24 did not alter the glomerular filtration rate. PMID- 9228197 TI - Influence of different dietary salts on the cardiovascular and renal effects of moxonidine in spontaneously hypertensive rats. AB - The influence of common salt (NaCl) and a novel potassium-, magnesium-, and L lysine-enriched mineral salt on the cardiovascular and renal effects of the selective imidazoline I1-receptor agonist moxonidine was examined in spontaneously hypertensive rats (SHR). Common salt was added at the level of 6% of the dry weight of the chow, and mineral salt at a 75% higher level of 10.5% thereof to produce the same NaCl concentration of 6% as in the common salt group. During the control diet an 8-week oral treatment with moxonidine (117 mg/1000 g of the dry weight of the chow producing an approximate daily dose of 10 mg/kg), lowered blood pressure by 13 mmHg. The common salt diet alone raised blood pressure by 27 mmHg. Moxonidine lowered blood pressure by 21 mmHg during the common salt diet, but the blood pressure remained 19 mmHg higher than in the moxonidine-treated SHR receiving the control diet (P<0.05). Unlike common salt, mineral salt alone did not raise blood pressure nor did it interfere with the antihypertensive effect of moxonidine. Moxonidine showed a kidney-protective effect during the control diet measured as decreased urinary protein excretion, but it did not affect the development of left ventricular hypertrophy. Moxonidine increased plasma renin activity during the control diet and it raised the serum aldosterone level both during the control and mineral salt diets. The vascular relaxation responses of the mesenteric arterial rings to both acetylcholine (an indicator of endothelium-dependent vascular relaxation) and nitroprusside and nitroprusside (an indicator of endothelium-independent vascular relaxation) were attenuated by the common salt diet alone but maintained during the moxonidine treatment. Our findings are consistent with the concept that moxonidine is able to improve the excretion of sodium. This effect might explain the maintenance of normal vascular relaxation during a high intake of common salt. These effects may partly account for the antihypertensive effect of moxonidine. PMID- 9228199 TI - Efflux studies allow further characterisation of the noradrenaline and 5 hydroxytryptamine transporters in rat lungs. AB - The aim of the present study was to further characterise the noradrenaline and 5 hydroxytryptamine [5-HT] transporters in rat lungs by examining the efflux of noradrenaline and 5-HT, respectively. Lungs from rats were isolated and perfused via the pulmonary artery. After loading the tissue with 3H-5-HT or 3H noradrenaline the efflux of the relevant amine from the lungs was examined for 15 25 min. The rate constant for efflux of 3H-5-HT increased by 81% when Na+ ions were removed from the perfusion solution; increased gradually when a selective 5 HT transporter inhibitor, 200 nM citalopram, was added to the perfusion solution for the final 6 min of efflux; and increased markedly and rapidly when substrates of the 5-HT transporter, tryptamine (18 microM) and 7-methyltryptamine (12 microM), were added for the final 6 min of efflux. These effects of the substrates were abolished by 1 microM citalopram, but were not significantly affected by 1 microM desipramine, a selective uptake, inhibitor. On the other hand, the previously described substrate-induced increase in the rate of efflux of noradrenaline was significantly reduced by desipramine but was unaffected by citalopram. The results show that efflux of 5-HT is mediated only by the 5-HT transporter, with no significant contribution of uptake1, and efflux of noradrenaline from rat lungs is mediated only by uptake1 and not by the 5-HT transporter. The effects of dopamine on the efflux of noradrenaline over a concentration range of 100-600 nM were investigated and the results showed that 50% of the maximal increase in the rate of efflux occurred at a concentration of 275 nM. This value did not differ from the Km for uptake of dopamine. This result implies that the only factor affecting the substrate-induced increase in noradrenaline efflux is the affinity of the substrate for uptake1. The efflux of noradrenaline was also examined in the absence and presence of two concentrations of desipramine (0.35 and 1.5 microM). Analysis of these results showed that uptake1 contributed approximately 81% and diffusion 19% to the total efflux of noradrenaline and that 90% of the total noradrenaline efflux was subject to reuptake by uptake1 into the pulmonary endothelial cells. PMID- 9228200 TI - Pharmacology and mode of action of bradykinin on mouse-isolated trachea. AB - In the present study, the effect of bradykinin on basal and precontracted mouse isolated trachea was investigated. In basal conditions mouse-isolated tracheal rings do not respond to bradykinin. However, when the tracheal rings were precontracted with carbachol (10(-7) M) a relaxation with bradykinin (3 x 10(-9) 3 x 10(-7)) was found. The maximal response amounted 69.7+/-4.1% (n=15) with a pD2 value of 7.2+/-0.21. The selective bradykinin B2 receptor antagonist HOE 140 (10(-10)-10(-8) M) antagonized the bradykinin-induced relaxation, while the bradykinin B1 receptor antagonist des-Arg9-Leu8-bradykinin (10(-6) M) had no influence. The selective bradykinin B1 receptor agonist des-Arg9-bradykinin (10( 6) M) caused a small relaxation (8.4+/-2.5%, n=6), which could be antagonized completely by the selective bradykinin B1 receptor antagonist des-Arg9-Leu8 bradykinin (10(-6) M) while addition of the selective bradykinin B2 receptor antagonist HOE 140 (10(-8) M) was without effect. In the presence of indomethacin (10(-6) M) the relaxation of bradykinin was completely abolished. Pretreatment of the tracheal rings with capsaicin, or the presence of the selective NK1 receptor antagonist RP 67851 (10(-6) M) or the presence of the nitric oxide synthase inhibitor L-NAME (3 x 10(-4) M) had no effect on the bradykinin-induced relaxation. In conclusion, these results demonstrate that the mouse-isolated tracheal is a preparation in which bradykinin exerts a relaxant response via stimulation of bradykinin B2 receptors. This response is probably mediated by prostaglandins. PMID- 9228201 TI - Tachykinin NK2 receptors: characterization in the rat small intestine by the use of a peptide agonist and a nonpeptide antagonist as radioligands. AB - The tachykinin NK2 receptors present in membranes of rat small intestine were characterized by means of tachykinin receptor agonists and antagonists, by using the natural agonist, NKA, or the nonpeptide antagonist SR 48968, as radioligands. The affinity of the antagonists was independent of the radioligand used, whereas the NK2 receptor selective agonists showed a different binding profile according to the radioligand. In particular, when using [125I]NKA, NKA and NKA(4-10) bound to a high affinity site, whilst [beta-Ala8]NKA(4-10) and GR 64349 bound to a high and a low affinity site; the high affinity site was still detected in the presence of 100 microM GppNHp which produced a strong inhibition of the specific binding of [125I]NKA. On the other hand, when using [3H]SR 48968, NKA bound to a high affinity site, [beta-Ala8]NKA(4-10) bound to a low affinity site and NKA(4 10) and GR 64349 bound to a high and a low affinity site; in this case, the high affinity site was no longer detected in the presence of 1 microM GppNHp, which did not reduce the specific binding of [3H]SR 48968 to NK2 receptors. We interpreted these data as an indication that in the rat small intestine membranes [125I]NKA labels multiple conformations of G protein-coupled NK2 receptor which are distinguished by the use of selective receptor agonists, but not by peptide or nonpeptide antagonists. PMID- 9228202 TI - Effects of gastroprokinetic agents on gastroparesis in streptozotocin-induced diabetic rats. AB - The influence of diabetic hyperglycemia on solid gastric emptying in rats was examined. Diabetes was produced by streptozotocin (STZ, 40 mg/kg i.v.), and diabetic hyperglycemia was observed from 1 day after the STZ injection. The gastric emptying of glass beads in the diabetic rats was significantly delayed compared with that in age-matched control rats at 1, 3 and 7 days after diabetes induction. A slight decrease in gastric emptying was observed in the diabetic rats from 2 to 52 weeks after the diabetes induction. We also investigated the influence of gastroprokinetic agents on STZ-induced diabetic gastroparesis and subdiaphragmatic vagotomy-induced gastroparesis in rats. The selective 5-HT3 receptor antagonists ramosetron (YM060), YM114 (KAE-393), granisetron and ondansetron, and the substituted benzamides (5-HT4 receptor agonist/5-HT3 receptor antagonists) cisapride mosapride and SC-53116 dose-dependently enhanced gastric emptying in normal rats. These compounds also reversed the impairment of diabetic gastroparesis rats at 7 days after the STZ injection, but higher doses were required. The solid gastric emptying in subdiaphragmatic vagotomized rats was also delayed. Ramosetron and the substituted benzamides cisapride and zacopride partially reversed the gastroparesis in the vagotomized rats. These results suggest that acute hyperglycemia is important mechanism for the delay of solid gastric emptying in diabetic rats. It is also suggested that selective 5 HT3 receptor antagonists and substituted benzamides enhance gastric emptying not only in normal rats but also in diabetic and vagotomized rats. PMID- 9228203 TI - The long-term sequelae of severe hypoglycemia on the brain in insulin-dependent diabetes mellitus. PMID- 9228204 TI - Troglitazone (CS-045): a new antidiabetic agent. PMID- 9228205 TI - Exercise training down-regulates ob gene expression in the genetically obese SHHF/Mcc-fa(cp) rat. AB - The recently cloned obesity gene (ob) encodes a protein, leptin, which is secreted from adipose tissue and interacts with hypothalamic receptors to decrease appetite, increase energy expenditure, and reduce body lipid stores. The levels of ob mRNA are increased in several models of obesity, consistent with the hypothesis that obese animals may be resistant to the actions of leptin. The present study examined the impact of increased energy expenditure through exercise training on ob mRNA gene expression and body composition in the SHHF/Mc fa(cp) male rat, a rodent model of obesity, insulin resistance, and type II diabetes. Six week old lean and obese animals were trained 8-12 weeks by treadmill running at 70% peak oxygen uptake, 5 days/wk, for 1.5 hr/day. After endurance training, exercised rats had significantly lower total body fat compared to sedentary rats of the same age, despite maintaining the same body weight. In the obese SHHF/Mcc-fa(cp) rat, the level of ob mRNA expression was markedly increased by four fold in subcutaneous adipose tissue compared to lean controls (p<0.05). In response to exercise training, there was a significant 85 % decrease in ob mRNA in exercised-training lean rats (p < 0.05) compared with non exercised controls, while in obese-exercised rats, ob gene expression was significantly reduced only by 50% relative to non-exercised obese rats (p < 0.05). These results demonstrate that exercise training reduces fat mass and ob mRNA in lean and obese rats, and supports the hypothesis of a feedback loop between the adipocyte and hypothalamus that attempts to maintain body weight at a constant level by reducing ob gene expression in response to increased energy expenditure. PMID- 9228206 TI - Long-term effect of prolactin treatment on glucose-induced insulin secretion in cultured neonatal rat islets. AB - Insulin secretion and 45Ca2+ uptake and efflux were studied in neonatal rat islets maintained in culture for 7 or 19 days in the absence or presence of prolactin (PRL). Insulin secretion in response to glucose (G), leucine (Leu), arginine (Arg) and carbachol (Cch) was augmented after 7 and 19 days in culture, compared to basal secretion (G 2.8 mM), in both PRL-treated and control islets. However, the increase in insulin secretion induced by the above secretagogues was higher in islets cultured in the presence of PRL for 19 days. In PRL-treated islets, the 45Ca2+ content after a 5 min incubation in the presence of G, Leu, Arg and Cch was significantly higher than the control only in islets cultured for 19 days. Except with Arg, the 45Ca2+ uptake in PRL-treated islets after a 90 min incubation was also significantly higher than the control only in islets cultured for 19 days. Finally, Leu-induced alterations in the 45Ca2+ efflux were higher in PRL-treated than in control islets cultured for 7 or 19 days. In the absence of external Ca2+, the reduction in 45Ca2+ efflux induced by glucose was also significantly higher in PRL-treated than in control islets. This effect was slightly potentiated after 19 days in culture. These data further support the hypothesis that PRL treatment enhances maturation of the secretory mechanism in neonatal islets. This effect can be potentiated even more if the treatment is prolonged. PMID- 9228207 TI - Effect of iodothyronines on electrophysiological properties of rat papillary muscle fibres. AB - We studied the effect of in vivo administration of 3,3',5-triiodo-L-thyronine (L T3) and other iodothyronines to thyroidectomized rats on the heart rate and electrophysiological properties, recorded in vitro, from papillary muscle fibres. The treatment with a daily substitutive L-T3 dose (3.7 nmoles/100 g body weight) for ten days was associated with a significant increase of heart rate and reduction of the action potential duration (APD) in comparison to that recorded from thyroidectomized rats. Analogous changes were obtained by treatment with equimolar dose of iodothyronines, such as 3,3',5-triiodo-D-thyronine (D-T3), 3,3',5,5'-L-tetraiodothyronine (L-T4), and 3'-isopropyl-3,5-diiodothyronine (3' Ip-T2), but not by treatment with 3,3',5'-L-triiodothyronine (L-rT3). Unlike the heart rate, APD was found in close correlation with the relative occupancy by iodothyronines of the heart thyroid hormone-specific nuclear sites reported in literature. Such a pattern was little modified when the stimulation rate of the preparations was increased from 1 Hz to 5 Hz, nearing the physiological rates. In fact, there were no significant APD changes for all groups, and only the significance of the APD difference between the T + L-T3 and T + L-rT3 groups was modified. In accordance to the expectations, L-T4 showed appreciable effects on APD at both frequencies tested, despite its small affinity for nuclear receptors. Indeed, it is well-known that a substantial proportion of the T4 effect derives from peripheral conversion to T3. Therefore, our results support the hypothesis that the long-term effects of thyroid hormone on ventricular electrophysiological properties are induced via synthesis of proteins that are part of ionic channels, through a pathway involving hormone interaction with the cell nucleus. PMID- 9228208 TI - Vitamin D receptor expression in chicken muscle tissue and cultured myoblasts. AB - Muscle has long been recognized as a target tissue for 1,25-dihydroxy-vitamin D3 (1,25[OH]2D3). Evidence of the presence of VDR is provided here, thus supporting the existence of a receptor-mediated mechanism of action of 1,25(OH)2D3. Vitamin D receptor (VDR) expression is evidenced by detection of VDR-mRNA, through reverse transcription and polymerase chain reaction (RT/PCR), in chicken muscle and muscle cells (myoblasts) as well as in a variety of tissues such as intestine, kidney, heart and brain. VDR presence is also demonstrated by Southern blot of PCR products with a specific VDR-cDNA probe and by immunocytochemistry carried out on myoblasts and cardiac myocytes. Localization of VDR is mainly nuclear and more faintly detected in the cytosol. PMID- 9228209 TI - Prostaglandin- and thromboxane-producing activity of isolated luteal cells from pseudopregnant rabbits. AB - The pseudopregnant rabbit is proposed as a suitable model for studies on physiology and endocrinology of the luteal phase. Pseudopregnancy is defined by corpus luteum function from day 1 to day 12 post hCG, as demonstrated by peripheral progesterone concentrations. The corpus luteum is highly vascularized which is extremely important to ensure progesterone secretion. Prostaglandins are potent vasoactive compounds and may be involved in controlling the ovarian/luteal blood flow. The present studies were designed to investigate the capacity of luteal cells for their intracellular production rates of prostaglandins in the early luteal phase. Pseudopregnancy was induced with a subcutaneous injection of FSH/LH, followed two days later by an intravenous injection of hCG, on days 0, 1 to 4 of pseudopregnancy luteal cells were isolated and incubated for 4 days. Media were collected every 24 h and analyzed for prostaglandins. The luteal cells were characterized by immunocytochemistry and progesterone measurements. Cultured luteal cells were able to convert exogenously applied arachidonic acid into PGI2, PGE2, PGF2alpha, and TXA2. The major compound that could be detected in the culture medium and in the cells was PGI2. The absolute values of the production pattern varied in all experiments in the ranges PGI2 > PGE2 > PGF2alpha > TXA2 with the greatest difference on day 3. In view of this fact the corpus luteum may contribute locally synthesized prostaglandins to regulate its own function. The physiological meaning of these findings should now be studied in a more optimal environment such as organ culture. PMID- 9228210 TI - Methylating activity in ovarian membranes changes during the estrous cycle. AB - The aim of this study was to characterize the methylation system of phosphatidylethanolamine (PE) in the ovarian membranes and identify their possible role on the ovarian physiology. The presence of two methylating activities was found, by using S-adenosyl-L-[methyl-3H] methionine (SAM) as methyl donor. One of them (Met I) uses PE as substrate, and the other one, (Met II), catalyses the two successive methylations of phosphatidyl-N monomethylethanolamine (PME) to phosphatidylcholine (PC). Met I shows a Km value of 5.71 microM and a Vm of 1.53 pmol/mg protein x min, working at pH 6.5, which seems to be the optimum. Met II exhibits low affinity for SAM, a Km of 50 microM, a Vm of 1.2 pmol/mg protein x min and it works an optimum pH of 9.5. Those enzymatic properties are in agreement with the amount of monomethylated and trimethylated products found working at pH 6.5 and 9.5, respectively. In order to investigate whether this enzyme system is affected or not by the hormonal environment influencing the ovary during the estrous cycle, the methylating activity was measured at its different stages. Both methylating activities were induced on proestrus but only Met I on diestrus. The total methylating activity increases when the ovarian membranes were obtained from rats injected with pregnant mare serum gonadotrophin (PMSG). Those results suggest that phospholipid methylation could be regulated by the hormonal environment during the estrous cycle. PMID- 9228211 TI - Circulating endothelin-1 in non-insulin-dependent diabetic patients with retinopathy. AB - Endothelin-1 (ET-1), a novel 21-amino acid vasoconstrictive peptide secreted by endothelial cells, has been thought to play a role in various forms of vascular disease. Diabetes mellitus is well known for its association with microvascular damage. To investigate whether ET-1 levels may be related to microangiopathy in diabetes mellitus, plasma ET-1 levels were measured in two groups of diabetic patients: A) 47 patients with non-insulin dependent diabetes mellitus (NIDDM) and retinopathy (28 M, 19 F; mean age 60.7+/-8.5 yrs) but without nephropathy (microalbuminuria < 30 mg/day) and hypertension (SBP < 140, DBP < 90 mmHg); group A was divided in three subgroups based on the severity of retinopathy: a) 16 with background retinopathy; b) 21 with pre-proliferative retinopathy; c) 10 with proliferative retinopathy. B) 8 patients with insulin-dependent diabetes mellitus (IDDM) recently diagnosed (6 M, 2 F; 16.4+/-3.8 yrs) without complications. C) 28 healthy subjects (HS) (16 M, 12 F; 47.8+/-11.8 yrs) as controls. In the NIDDM group the ET-1 concentration was significantly higher (17.3+/-2.4 pg/ml) than both in the HS (8+/-4.7 pg/ml) and IDDM patients (10.2+/-3.7 pg/ml) (p < 0.0001). In the subgroups with retinopathy the ET-1 levels were a) 15.1+/-4.3 pg/ml; b) 22.2+/-6.8 pg/ml and c) 16.6+/-5.1 pg/ml. These values were significantly elevated as compared to HS (p<0.001; p < 0.0001; p < 0.002, respectively), being the highest levels of ET-1 observed in the NIDDM patients with pre-proliferative retinopathy. In conclusion our study revealed that the ET-1 concentrations are elevated in NIDDM patients with retinopathy especially in those patients with pre proliferative retinopathy. PMID- 9228212 TI - Reverse 3,3',5'-triiodothyronine (rT3) enhances hyperglycemic and lipemic effects of heat-stress in chickens. PMID- 9228213 TI - Minocycline induces an increase in the number of excreting pilosebaceous follicles in acne vulgaris. A randomised study. AB - The effects of treatment with minocycline 100 mg per day on sebaceous excretion in acne vulgaris using lipometry and Sebutape were studied in 45 patients in an open study as well as in a randomised placebo-controlled study. In both studies a subclinical increase in sebaceous excretion was noted from the 28th day of treatment. This effect continued for 1 month after the end of treatment. The increase in sebaceous excretion was concomitant with an increase in the number of excreting pilosebaceous follicles. Minocycline may cause the follicles to become unblocked by acting on the factors responsible for ostial hyperkeratosis, in addition to an antibacterial effect on retentional acne lesions. PMID- 9228214 TI - Substance P binding to peripheral blood mononuclear leukocytes in atopic dermatitis. AB - Substance P has various immunomodulatory effects, including in vitro modification of lymphocyte proliferation and cytokine release. Elevated levels of substance P and increased staining of substance P-positive nerve fibres have been reported in atopic dermatitis patients. We examined fluoresceinated substance P binding to a range of lymphocyte subsets and compared the results in atopic dermatitis, non atopic psoriasis patients and normal controls. Fluoresceinated substance P and phycoerythrin-labelled monoclonal antibodies to CD3, CD4, CD8, CD57, CD19 and CD14 were incubated in duplicate with Ficoll-Hypaque separated peripheral blood mononuclear leukocytes. With flow cytometry the fluoresceinated substance P positive cells were identifiable as a peak of positively fluorescent cells, and the percentages of positive cells were measured. We have demonstrated binding of fluoresceinated substance P to all subsets examined, with significantly less binding to atopic dermatitis CD3-, CD8- and CD57-positive cells. This may affect cytokine release and hence be important in the pathogenesis of atopic dermatitis. PMID- 9228215 TI - Terpene-enhanced transdermal permeation of water and ethanol in human epidermis. AB - The study was performed to investigate the effect of penetration enhancers on the stratum corneum barrier. Epidermal membranes were prepared from freeze-stored ( 70 degrees C) Caucasian breast skin and mounted in a flow-through diffusion cell. The validity of the freeze storage procedure was verified by measurement of [3H] water penetration. The effect of the cyclic terpene, carveol, on the transdermal penetration of water and ethanol was studied in vitro. Control ethanol and water penetration measured with a donor solution of 50% ethanol/PBS (w/w) was 1.9+/-0.2 and 3.6+/-0.5 x 10(-3) cm/h. The addition of 3% carveol to the donor solution increased the permeation of ethanol and water after 4 h to 8.3+/-1.1 and 12.5+/ 1.9 x 10(-3) cm/h, respectively. In a separate experiment, terpinen-4-ol and alpha-terpineol were also tested, in addition to carveol, for effect on tritium flux. No significant difference in maximum tritium flux was obtained between the three terpenes studied. The maximum increase in permeability coefficients of carveol, terpinen-4-ol and alpha-terpineol was 10.6, 8.7 and 10.9, respectively. PMID- 9228216 TI - Topical calcitriol (1,25-dihydroxyvitamin D3) treatment of psoriasis: an immunohistological evaluation. AB - The potent calciotropic hormone calcitriol (1,25-dihydroxyvitamin D3, 1,25(OH)2D3) has been shown to be very effective and safe in the topical treatment of psoriasis. In vitro, 1,25(OH)2D3 inhibits proliferation and stimulates differentiation of human keratinocytes. Increasing evidence suggests an immunoregulatory function of this potent steroid hormone. To further characterize the biological effects of topical calcitriol treatment in psoriasis, we have analyzed immunohistochemically the expression of markers for epidermal proliferation (proliferating cell nuclear antigen=PCNA) and differentiation (transglutaminase K, involucrin, cytokeratin 16), as well as inflammation (CD1a, 55 kDa TNF-receptor, NAP-1/IL-8) in calcitriol-treated psoriatic skin in situ. Our findings strongly support the hypothesis that calcitriol modulates keratinocyte proliferation/differentiation as well as inflammation in human skin in vivo. The immunoreactivity of markers for epidermal proliferation and differentiation, as well as of CD1a and NAP-1/IL-8, changed after 8 weeks of calcitriol treatment almost completely to the pattern characteristic for non lesional psoriatic skin, while a large number of 55 kDa TNF-receptor positive cells could be found in the dermal compartment. PMID- 9228217 TI - Evidence that HLA-Cw6 determines early onset of psoriasis, obtained using sequence-specific primers (PCR-SSP). AB - Psoriasis vulgaris has previously been shown to associate with certain HLA alleles. HLA-Cw6 is considered to be the primary association, based on calculations of relative risk after serological typing. This association is reportedly more pronounced in early- than in late-onset psoriasis. We performed a PCR-based typing with sequence-specific primers, which has been shown to give a more complete result than serology. Two hundred and one unrelated patients with psoriasis, with a mean age of 40 years, and 77 healthy controls were typed. Two thirds (67%) of the patients were positive for one or two copies of the allele, while the corresponding figure for the control group was 12%. A significant peak for age at onset of 21 or younger was seen for the Cw6 carriers. For patients older than 21 at onset, the frequency of Cw6 was significantly lower; e.g. for patients with an age at onset between 30 and 35 the frequency was comparable to the level of the control group. The high frequency of Cw6 among patients with an age at onset of 21 or younger is in agreement with data of other groups. In comparison with this age-at-onset group the frequency of Cw6 is sharply reduced among patients with an age at onset of 22 years or older, which contrasts with earlier studies. This may reflect differences between population groups but may also be due to the higher sensitivity of the PCR-based HLA-Cw6 typing method. In view of these findings, we suggest that psoriasis is a genetically determined disease, in which the additional presence of HLA-Cw6 is associated with the characteristic of early onset. PMID- 9228218 TI - Dominant dystrophic epidermolysis bullosa albopapuloidea Pasini -- ultrastructural observations of albopapuloid lesions and a type VII collagen DNA polymorphism study of a family. AB - We describe a case of dominant dystrophic epidermolysis bullosa (DDEB) albopapuloidea Pasini. The patient was a 42-year-old female with albopapuloid lesions on her back, which had developed when she was 17. Histological examination of the albopapuloid lesions showed prominent proliferation of immature collagen bundles and deposition of amorphous material, which stained positively with Alcian blue. Electron microscopy of her albopapuloid lesions revealed marked nodular proliferation of collagen bundles from just beneath the basal lamina to the mid-dermis. In the light of these findings, we speculate that such albopapuloid lesions result from a reactive accumulation of collagen and glycosaminoglycan occurring on the EB skin. A PvuII and AluI polymorphism study of type VII collagen DNA from the patient's family suggests that the candidate gene for DDEB in her pedigree could be the type VII collagen gene. PMID- 9228220 TI - Cutaneous adverse reaction to ciprofloxacin: demonstration of specific lymphocyte proliferation and cross-reactivity to ofloxacin in vitro. AB - Ciprofloxacin (CPFX) is a widely used fluoroquinolone antibiotic, inducing cutaneous adverse drug reactions in about 1 to 2% of the treated patients. Conclusive diagnosis of drug allergy, however, still remains a major problem in daily clinical practice. Here, we present 2 patients with drug allergy to CPFX. In both cases the clinical suspicion for CPFX as the causative agent was confirmed in vitro by means of the lymphocyte transformation test, whereas epicutaneous patch tests remained negative. In vivo, a small percentage of the drug is biotransformed to the three major metabolites desethylene-, sulfo- and oxociprofloxacin. Though structurally closely related to their mother compound, these metabolites failed to induce in vitro lymphocyte proliferation in both patients. On the other hand, in vitro crossreactivity to ofloxacin, another fluorinated quinolone, could be demonstrated, which to our knowledge has not previously been reported. PMID- 9228219 TI - Insulin inhibits tyrosinase activity and 5-S-cysteinyldopa formation in human melanoma cells. AB - The effects of insulin on melanogenesis were examined in human Swift melanoma cells. When these cells were grown in a chemically defined culture medium containing insulin (5 microg/ml), they showed a low pigmentation in association with a high activity of glutathione peroxidase (GPO) and a low activity of tyrosinase. In Eagle's minimum essential medium supplemented with foetal calf serum (EMEM-FCS), the Swift cells showed an intense pigmentation in association with a low GPO activity and a high tyrosinase activity. Modulation of GPO activity with sodium selenite had no effect on melanogenesis variables. In contrast, addition of insulin (5 microg/ml) to the EMEM medium led to a marked decrease in tyrosinase activity (p<0.001) and to a concomitant reduction in the levels of 5-S-cysteinyldopa (p <0.01). These results indicate that insulin inhibits the formation of 5-S-cysteinyldopa and that of melanin via the inhibition of tyrosinase activity. PMID- 9228221 TI - Paraneoplastic pemphigus associated with pancreatic carcinoma. AB - A case of paraneoplastic pemphigus associated with pancreatic carcinoma is presented. The histopathological and immunological features of the case, which are consistent with and differ from the accepted diagnostic criteria, are discussed. PMID- 9228222 TI - Rapid cytological diagnosis of primary skin tumours and tumour-like conditions. AB - This study presents the results of fine needle aspiration cytology performed on 1,263 skin lesions which were clinically suspicious for neoplasia. The purpose of the study was to investigate the accuracy of fine needle aspiration cytology for the diagnosis of skin tumours and to assess its clinical value. Twenty-one to 27 Gauge needles were used and the specimens were stained by a quick Giemsa stain. The cytological examination reported 826 primary malignant tumours and 437 benign lesions. Five hundred and thirteen of the cytologically malignant cases and 123 of the benign ones had a subsequent histological examination. The correlation between cytology and histology revealed 6 false positive cytological results and one false negative. Persuaded by our results, we believe that fine needle aspiration cytology can give highly reliable information concerning the histological type or primary skin tumours. It can also detect or exclude relapses of previously treated neoplasms. The procedure is non-traumatic, safe, quick, inexpensive and very well tolerated by the patients. PMID- 9228224 TI - No evidence for Borrelia burgdorferi infection in lesions of morphea and lichen sclerosus et atrophicus in Spain. A prospective study and literature review. AB - The possible association of Borrelia burgdorferi with morphea and lichen sclerosus et atrophicus has been the focus of research and discussion in dermatology during the last 10 years. To investigate the etiopathogenic role of B. burgdorferi in morphea and lichen sclerosus et atrophicus lesions in Spain, we studied 14 cases: 8 patients with lichen sclerosus et atrophicus and 6 with morphea. For the whole group, a prospective study was performed, including serologic studies by indirect immunofluorescence, histologic evaluation of skin biopsy specimens, culture studies, and polymerase chain reaction with different primers sensitive for detecting virtually all B. burgdorferi strains tested to date. Although one patient with morphea had positive serologic findings at low titer, we were not able to culture or detect borrelial DNA in any of the specimens. These findings do not confirm an association between B. burgdorferi and morphea and lichen sclerosus et atrophicus. PMID- 9228226 TI - Refractory ulcerative lupus vulgaris associated with CD4 lymphocytopenia, inversion of chromosome 14, primary amenorrhoea and mental retardation. AB - A case of ulcerative lupus vulgaris, confirmed by polymerase chain reaction (PCR) is reported. The initial lesion of our case was a papule on the nose, which progressed during antituberculous treatment and caused cartilage destruction and ectropion. Immunological analysis revealed CD4 lymphocytopenia, and the possibility of idiopathic CD4 lymphocyte deficiency was considered. In addition, the patient had primary amenorrhoea, mental retardation and inversion of chromosome 14. CD4 lymphocytopenia and chromosomal abnormality are the possible causes of antituberculous treatment failure. PMID- 9228225 TI - Markers in cutaneous lupus erythematosus indicating systemic involvement. A multicenter study on 296 patients. AB - Lupus erythematosus (LE) is an autoimmune disorder, involving the skin and/or other internal organs. As cutaneous variants, chronic discoid LE (CDLE) and subacute cutaneous LE (SCLE) usually have a better prognosis, however, involvement of internal organs with transition into systemic disease may occur. The aim of this study was to assess the significance of some clinical and laboratory criteria that could serve as markers for early recognition of systemic involvement in cutaneous LE. Three hundred and seventy-nine patients with LE, seen in five cooperating Departments of Dermatology during the years 1989-1994, were documented by electronic data processing according to a common protocol. Two hundred and forty-five of these patients had cutaneous LE (CDLE or SCLE), and 51 had systemic LE (SLE) and were included in this study. Forty-nine patients with either CDLE/SCLE or SLE were not evaluated because of incomplete documentation; also, 34 patients suffered from other LE subsets and were likewise excluded from the evaluation. Multivariate statistical analysis was used to assess the value of seven selected variables for distinguishing between the CDLE/SCLE and SLE groups: ESR, titers of antinuclear antibodies, anti-dsDNA-antibodies, photosensitivity, presence of arthralgias, recurrent headaches and signs of nephropathy. Univariate and multivariate analysis of the obtained data showed that signs of nephropathy (proteinuria, hematuria) was the variable with the highest statistical relevance for distinguishing between patients with cutaneous (CDLE/SCLE) and with systemic LE (SLE) in all statistical models tested, followed by the presence of arthralgias and of high ANA titers (> or =1:320). In contrast, low ANA titers as well as anti-dsDNA antibodies showed little or no statistical relevance as a criterion for distinction. It seems, therefore, that cutaneous LE patients showing signs of nephropathy, presence of arthralgias and elevated ANA titers (> or =1:320) should be carefully monitored, because they may be at risk of developing systemic LE involvement. PMID- 9228223 TI - Stress and alopecia areata: a psychodermatologic study. AB - Psychosocial stress has been reported to play a role in the onset and/or exacerbation of alopecia areata. Little is known about the clinical characteristics of alopecia areata patients whose alopecia is stress-reactive. We examined the relation between the stress reactivity of alopecia areata and a wide range of psychosocial measures among 16 patients with alopecia areata/totalis and 28 patients with alopecia universalis. The degree to which the alopecia was exacerbated by stress was measured by patient ratings on a 10-point scale. A wide range of psychologic measures correlated (p<0.05) with the stress reactivity score. Stepwise logistic regression analysis revealed that patients with higher depression scores were more likely to be in the high-stress reactor group. Patients whose alopecia is stress-reactive may suffer from depressive illness, a potentially important consideration in the overall management of such patients. PMID- 9228227 TI - Low-frequency ultrasound treatment of chronic venous leg ulcers in an outpatient therapy. AB - Low-dose ultrasound seems to be an effective method to enhance wound healing, particularly in chronic venous leg ulcers. The aim of our investigation was to examine the effect of 30 kHz low-dose ultrasound in local treatment of chronic venous leg ulcers, when added to conventional therapy of outpatients. Twenty-four patients with chronic ulcerations of the leg due to chronic venous insufficiency were randomised in placebo-controlled parallel groups in a single-blind clinical study. Patients were randomised to conventional therapy with topical application of hydrocolloid dressings and compression therapy or conventional therapy with additional ultrasound treatment for 12 weeks. The ultrasound treatment consisted of 10 min of footbathing, with application of 30 kHz continuous ultrasound 100 mW/cm2 three times a week. The ulcer area was measured by planimetry, using a millimeter grid before treatment and after 2, 4, 6, 8, 10 and 12 weeks of therapy. The ulcer radius and the daily ulcer radius reduction were calculated. Colour photographs of the ulcers were taken under standard conditions at the same time. After each treatment local findings and side effects were recorded. After 12 weeks of treatment the control group showed a mean decrease of 16.5% in the ulcerated area. In contrast the mean ulcerated area decreased by 55.4% in the ultrasound group (p<0.007). The daily ulcer reduction in the ultrasound-treated patients was 0.08 mm+/-0.04 mm and in the placebo patients 0.03 mm+/-0.03 mm. Patients recorded only minor side-effects such as a tingling feeling and occasionally pinhead-sized bleeding in the ulcer area. The application of low frequency and low-dose ultrasound is a helpful treatment option in chronic venous leg ulcers, especially if they do not respond to conventional ulcer treatment. PMID- 9228229 TI - Semi-quantitative measurements of body hair in hirsute women compare well with direct diameter measurements of hair shafts. AB - No standards exist for the evaluation of hair in hirsute women. This study compared the semi-quantitative visual scoring of body hair, using the Ferriman & Gallwey scale, in 88 hirsute women with direct objective measurements of hair shaft diameter and daily linear growth rates of hair growing on the pre-auricular area of the face, the forearm, the anterior abdominal wall and the anterior thigh. There was a significant correlation between the semi-quantitative score and diameter measurements on the forearm, abdominal wall and thigh. There was no relationship between linear growth rates at any of the four sites and the semi quantitative score. The conclusion of this report is that suitably standardised and controlled semi-quantitative measurement of hair in hirsute women with visual analogue scores would appear to offer information similar to that obtained by direct measurement of hair diameter. PMID- 9228228 TI - Effect of ebastine on mosquito bites. AB - Mosquito bites usually cause wealing and delayed bite papules. Cetirizine decreases wealing, bite papules and pruritus but the effect of other antihistamines on mosquito bites is unknown. We studied the effect of ebastine in 30 mosquito bite-sensitive adult subjects. Ebastine 10 mg or 20 mg and placebo were given for 4 days in a cross-over fashion. Aedes aegypti bites were given on forearms. The size of the bite lesions and pruritus (visual analogue score) were measured at 15 min, 2, 6, and 24 h after the bites. Twenty-five subjects were evaluable in the study. At 15 min ebastine decreased significantly the size of the bite lesion (p = 0.0017) and pruritus (p<0.0001). The effects of 10 mg and 20 mg of ebastine were similar. No significant effect was found at 2, 6 or 24 h, but when the measurements at all four time points were compiled the size of the bite lesion and pruritus score decreased significantly. Sedation occurred during ebastine treatment in 6 (21%) and during placebo treatment in 2 (7%) subjects. The present results show that prophylactically given ebastine is effective against immediate mosquito bite symptoms. PMID- 9228230 TI - Is vulvar vestibulitis an inflammatory condition? A comparison of histological findings in affected and healthy women. AB - Vulvar vestibulitis, as defined by Friedrich, is considered to be inflammatory, despite the fact that the normal histology of this specific area has previously not been characterized. The aim of the present study was to compare the normal histology of the vulvar vestibulum with findings in localized vulvar vestibulitis. Biopsies were taken at the area of the vestibulitis, i.e. at the openings of the Bartholin's duct. Eleven control specimens were examined histologically and compared to 24 specimens obtained from 20 patients. All samples were also tested for human papillomavirus, and they were all negative. In control specimens, as well as in specimens from patients, subepithelial inflammatory cells, sometimes aggregated into lymph follicles and/or small groups of lymphocytes were found. The conclusion is reached that the occurrence of inflammatory cells in vestibular tissue is a normal finding and cannot serve as a histological indicator of vulvar vestibulitis. PMID- 9228231 TI - Urticaria-like follicular mucinosis in a young female patient. PMID- 9228232 TI - Cutaneous leukocytoclastic vasculitis in a case of ankylosing spondylitis. PMID- 9228233 TI - Substance P is not involved in primary and secondary erythermalgia. PMID- 9228234 TI - Localized crusted scabies in a patient with acquired immunodeficiency syndrome. PMID- 9228235 TI - Immunohistochemical detection of cathepsins in Merkel cell carcinomas. PMID- 9228236 TI - Kawasaki syndrome or atypical measles mimicking Kawasaki syndrome? PMID- 9228237 TI - Urethritis associated with isotretinoin therapy. PMID- 9228238 TI - "Complex regional pain syndrome" after trauma from high-heeled shoe. PMID- 9228239 TI - The simultaneous occurrence of Hailey-Hailey disease, Graves' disease and multiple sclerosis in the same patient. PMID- 9228240 TI - Hair loss in children on long-acting gonadotropin-releasing hormone agonist triptorelin treatment. PMID- 9228241 TI - Autosomal dominant cornea plana: clinical findings in a Cuban family and a review of the literature. AB - Cornea plana may occur in connection with malformations of the eye or of other parts of the body. As an isolated ocular anomaly, it may be inherited in an autosomal recessive or in an autosomal dominant fashion. We have previously mapped genes for both forms of the disease to 12q21. We studied 36 members of three generations of a Black Cuban family with autosomal dominant cornea plana. Three affected males and 11 affected females were examined. Corneal refraction varied between 37.50 and 42.75 diopters. Horizontal corneal diameter ranged from 8.75 to 11.25 mm. The cornea was clear and the limbal zone only occasionally widened. A marked arcus senilis was present in six patients aged 30 to 58 years, but in none of their healthy relatives. The anterior chamber was shallow in those affected, varying in depth from 1.68 to 2.38 mm. One woman was blind from closed angle glaucoma. The axial length was within normal limits in all patients. PMID- 9228242 TI - A mild phenotype of autosomal dominant retinitis pigmentosa is associated with the rhodopsin mutation Pro-267-Leu. AB - By screening blood samples from patients with autosomal dominant retinitis pigmentosa, we found in one of the families a rhodopsin mutation (Pro-267-Leu), which segregates with the disease in two affected and five unaffected family members. Here, we present the results of the clinical evaluation of the family, including full-field electroretinography from the two affected family members. A 25-year-old family member with the mutation had an almost normal electrophysiological retinal response. The patient's father, who was also heterozygous for the mutation and had mild subjective symptoms of retinitis pigmentosa, demonstrated a substantially preserved retinal function. Our results suggest that the Pro-267-Leu rhodopsin mutation is associated with a very mild phenotype of retinitis pigmentosa. Young patients with the disease may have minimal pathological changes in the electroretinogram and some patients with few symptoms may be affected without acquiring a diagnosis of eye disease. PMID- 9228243 TI - Histopathological and immunohistochemical findings associated with a null mutation in the Norrie disease gene. AB - PURPOSE: To determine the clinical, histopathological, and immunohistochemical ocular changes associated with a null mutation in the Norrie disease protein (NDP) gene. METHODS: Tissue from a six-month-old boy with bilateral retrolental membranes and retinal detachment was obtained during vitreoretinal surgery. Histological sections were stained immunohistochemically with specific antibodies. No eye diseases with severe visual impairment or blindness were reported in the parents and their families. The NDP gene was analyzed by standard molecular genetic methods. RESULTS: A severe reduction in the number of retinal ganglion cells and a largely disarranged and hypoplastic inner nuclear layer were visible in the tissue specimen. Areas of the tissue with advanced pathology displayed massive fibrovascular proliferation in the vitreous cavity. Shrinkage and traction resulted in folding and detachment of the outer retina. Immunohistochemical reactivity for MIB(1) antigen demonstrated many proliferating cells in the vitreous, but no proliferative activity in the neuroretina. Retinal neurons showed a high grade of differentiation and expressed uniformly neuron specific enolase and synaptophysin. A 1-base pair insertion (544/545insA) in the NDP gene was found in the affected boy. This mutation predicts a 'functional null allele' due to a shift in the reading frame and, thus, a premature termination of mRNA translation after 55 instead of 133 amino acids. CONCLUSIONS: Loss of function of the NDP gene causes marked hypoplasia of the inner retinal cell layers and fibrovascular proliferation in the vitreous cavity, leading to retinal folding and detachment. The NDP therefore seems to play a critical role in terminal differentiation of the inner retinal cell layers and establishment and maintaining of anti-proliferative cellular interactions in the vitreous. PMID- 9228244 TI - Wilson's disease: presymptomatic patients and Kayser-Fleischer rings. AB - PURPOSE: We evaluated patients with Wilson's disease to determine (1) whether presymptomatic patients who have Kayser-Fleischer (K F) rings demonstrate a more significant alteration of copper metabolism than those who do not have K F rings, and (2) whether presymptomatic patients have smaller K F rings than symptomatic Wilson's disease patients. METHODS: Thirty-two patients with presymptomatic Wilson's disease were retrospectively analyzed. Sixteen of these had received no prior anti-copper therapy and underwent testing for baseline copper metabolism (24-hour urine copper, liver copper, and plasma ceruloplasmin). Quantitative measurements of K F rings were made for the group of untreated presymptomatic patients and a control group of symptomatic Wilson's disease patients. RESULTS: We hypothesized that the 24-hour urine copper, in particular, would correlate with the presence of a K F ring. However, no significant difference was found between any of the baseline copper variables for presymptomatic patients who had K F rings compared to those who did not. K F rings of presymptomatic patients were found to be significantly smaller than K F rings of patients with symptomatic Wilson's disease (p < 0.05). CONCLUSIONS: While this study does not show any relationship between urinary copper excretion and the presence of K F rings, it suggests that the larger K F ring size correlates with Wilson's disease severity. PMID- 9228245 TI - A further observation of corneal dystrophy and perceptive deafness in two siblings. AB - We studied two siblings with the rare association of corneal dystrophy and perceptive deafness (Harboyan syndrome). To our knowledge, this is the third description of this hereditary disorder. The results of the clinical, genetic, audiometric, and ocular examination of the two siblings and the type of inheritance, which agree with the previous description of the syndrome, are reported. Various hereditary syndromes associated with corneal dystrophy are reviewed. PMID- 9228246 TI - Peroxisomal bifunctional enzyme deficiency with associated retinal findings. AB - Peroxisomal disorders include single enzyme defects and defects of peroxisomal fatty acid oxidation enzymes. Peroxisomal bifunctional enzyme complex deficiency is a recently recognized abnormality of fatty acid metabolism. We present one patient with peroxisomal bifunctional enzyme deficiency in association with a flecked retina. This clinical association has only been previously reported once. The finding of a flecked retina in an infant presenting with hypotonia, seizures, and failure to thrive is highly suggestive of peroxisomal bifunctional enzyme complex deficiency. PMID- 9228247 TI - Optic neuropathy associated with mitochondrial tRNA[Leu(UUR)] A3243G mutation. AB - PURPOSE/BACKGROUND: To report the association of optic neuropathy and mitochondrial tRNA[Leu(UUR)] A3243G mutation which is known to be responsible for MELAS (mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes), diabetes mellitus with deafness, and progressive external opthalmoplegia. Pigmentary retinopathy, opthalmoparesis, and ptosis have been relatively frequently reported to be associated with the mutation in the literature. However, optic atrophy has rarely been reported to be associated with the mutation. METHODS: Analyses including measurement of the corrected visual acuity, color vision, pupillary examination, funduscopic examination, visual field, visual evoked potential, and brain imaging study were performed in our two patients with the mutation. RESULTS: In disagreement with previous reports, this study revealed the association between optic neuropathy and the mutation in the two patients. CONCLUSION: There might be some degree of optic neuropathy related to the tRNA[Leu(UUR)] A3243G mutation. Thus more detailed ophthalmologic examination should be done to detect optic neuropathy. PMID- 9228248 TI - Morphology and diagnostics of human toxoplasmosis. AB - Toxoplasmosis has regained clinical importance. In addition to intrauterine fetal infections, the acquired toxoplasmosis endangers an increasing number of immunocompromised adults. This disease affects not only oncologic patients, but also organ transplant recipients and, above all, AIDS patients. Toxoplasmosis has five clinical subtypes: lymphadenopathy, encephalitis, chorioretinitis, disseminated toxoplasmosis and congenital toxoplasmosis. In biopsies of different organs and autopsies of persons who died as a result of an infectious disease, the pathologist is involved in the diagnostic procedures. The findings of the parasites in tissue specimens still constitute the morphologic feature of toxoplasmosis. Differential diagnosis has been improved by immunohistologic methods for demonstrating toxoplasmas in tissue specimens and by the use of polymerase chain reaction for identification of toxoplasmatic antigens. This review describes the pathomorphology and possibility regarding the diagnosis of toxoplasmosis. PMID- 9228250 TI - p53 protein and tumorigenesis in the uterine cervix. AB - The relationship between molecular abnormalities of p53 tumor suppressor gene product and cancer has been well documented. That correlation may exist between immunocytochemically detectable amount of p53 protein and neoplasia is evidenced by several studies. Detection of p53 protein by immunocytochemistry varies depending on the methods and antibodies used. It has been suggested that the quantitative aspect of p53 protein expression and the proportion of cells expressing p53 may be of clinical importance in human malignancies. In the present study, we have examined the expression of p53 protein in various grades of lesions of the uterine cervix. Statistical analysis showed a good correlation between expression of p53 protein and histologic grade of lesions. Increased expression of p53 in dysplastic and malignant lesions compared to non dysplastic lesions suggests that p53 protein accumulation may be an early event in carcinogenesis. PMID- 9228249 TI - Expression of c-erbB-2 oncogene and p53 tumor suppressor gene in benign and malignant breast tissue: correlation with proliferative activity and prognostic index. AB - The expression of c-erbB-2 oncogene and p53 tumor suppressor gene was studied in methacarn-fixed, paraffin-embedded biopsy specimens from 58 benign breast lesions and 129 sporadic breast carcinomas, using the supersensitive monoclonal antibodies CB 11 and BP 53-12-1 and the biotin-streptAvidin-amplified methodology. None of the benign lesions studied, which included 36 fibrocystic lesions with mild or florid epithelial hyperplasia, 12 fibrocystic lesions with ADH or ALH and 10 fibroadenomas, demonstrated membrane staining for c-erbB-2 or nuclear immunoreactivity for p53. Overall, 48.06% of primary breast carcinomas showed membrane expression of c-erbB-2, while 28.68% were p53 positive. Those showing p53 immunoreactivity displayed a nuclear and/or cytoplasmic staining type. A significant correlation was seen between c-erbB-2 and p53 expression (r = 0.213, p < 0.05), as well as between c-erbB-2 status and PSNA score (r = 0.221, p < 0.05). In addition, c-erbB-2 and p53, separately or in combination, correlated significantly with the prognostic index. In conclusion, immunohistochemistry of c erbB-2 and p53 immunohistochemistry allows a better definition of intraductal proliferative lesions and assists in the differentiation between ADH and DCIS. It provides additional clues with regard to the biologic behavior of invasive ductal carcinomas (NOS and medullary). PMID- 9228251 TI - Immunohistochemical expression of p53 and c-erbB-2 in breast carcinoma: relation with epidemiologic factors, histologic features and prognosis. AB - We have performed immunohistochemical staining for p53 and c-erbB-2 on formaldehyde-fixed, paraffin-embedded primary invasive ductal carcinomas from 112 patients, with a minimal follow-up time of 60 months. All of them had received postoperative chemoradiation therapy. We have analyzed the association of these factors with epidemiologic risk factors, histopathologic features and hormonal receptor status and the influence on prognosis. Our results indicate that the expression of c-erbB-2 protein defines a group of node-negative patients with poor prognosis. The overexpression of c-erbB-2 has shown a significant association with estrogen receptor status (those tumors expressing c-erbB-2 are usually estrogen receptor negative), presence of fibrosis and lymphoplasmacytoid infiltrates. P53 expression has shown no relation either with prognosis or with any other histopathologic or clinical feature. The only factors with prognostic influence in our series have been tumor size, the presence of node metastases, TNM stage and the prognostic morphometric index (Baak's index), apart from c-erbB 2 in node-negative patients. However, only the TNM stage showed an independent association with prognosis after a multivariate analysis. In summary, in our experience the expression of p53 protein has no prognostic influence on breast carcinoma, and TNM stage remains to be as the most powerful prognostic factor in these patients. PMID- 9228252 TI - The basement membrane and tumor progression in the uterine cervix. AB - Immunocytochemical localization of the basement membrane (BM) proteins laminin, type-IV collagen and fibronectin were analyzed in normal cervical epithelium, low grade squamous intraepithelial lesions (SILs), high grade SILs and invasive squamous cell carcinoma (SCC) of the uterine cervix. A regular, thick and continuous BM was present in normal cervical epithelium and low grade SIL. Interruptions and discontinuity of the BM were more evident in high grade SILs. There was a good correlation between increasing severity of the lesion and increasing number of breaks. In SCC, the distribution of laminin, collagen IV and fibronectin was related to the degree of cellular differentiation, with decreased immunoreactivity being evident in moderately and poorly differentiated tumors. As the invasive potential of the tumor increased, the fragmentation and loss of BM was more evident. Fibronectin showed only moderate to mild immunoreactivity in normal cervical epithelium and low grade SILs. However, the intensity of expression increased in high grade SILs especially in the peritumoral stroma. It may therefore be concluded from these results that snythesis and reabsorption of BM proteins may be related to shifts in cellular metabolism during tumorigenesis. PMID- 9228253 TI - Lupus and nonlupus membranous glomerulopathy. Quantitative comparison of the subepithelial deposits and glomerular basement membrane including clinicomorphologic correlations. AB - We examined quantitatively 11 renal biopsy specimens from patients with class Va WHO lupus membranous glomerulopathy (LMGN) and 16 from patients with primary (nonlupus) membranous glomerulopathy (NLMGN) for whom both light and electron microscopy as well as immunofluorescence microscopy and full clinical data were available and compared these specimens with six cases of normal controls. In LMGN, subepithelial deposits resembled those seen in stage III of membranous glomerulopathy (MGN) according to the scheme proposed by Churg's group, i.e., for the present study only advanced cases of NLMGN (stage III according to this scheme) were selected. The electron micrographs were scanned in Primax flatbed A4 scanner and morphometric investigations were then performed by means of a computer image analysis system to compare glomerular basement membrane (GBM) thickness and the electron-microscopic density of the deposits in LMGN and NLMGN as well as to study whether these parameters could correlate with the clinical data. The study revealed that in LMGN the GBM thickness and the electron microscopic density of the deposits were significantly increased in comparison with NLMGN. It should also be noted that both in LMGN and NLMGN groups the degree of proteinuria was closely correlated with the density of the deposits, but not with the GBM thickness. Moreover, no correlations were found between serum creatinine and the GBM thickness as well as between serum creatinine and the density of the deposits in these groups. In conclusion, the present data confirm that in LMGN and NLMGN proteinuria mainly depends on density of the subepithelial deposits. Furthermore, in cases with especially high density of these deposits systemic lupus erythematosus (SLE) should be taken into consideration, even if this etiology was not clinically suggested at the time of biopsy. PMID- 9228254 TI - Computerized determination of proliferating cell nuclear antigen expression in meningiomas. A comparison with non-automated method. AB - Proliferating cell nuclear antigen (PCNA) expression has been proven to be a significant marker of cell proliferation in meningiomas, which correlates with growth rate and, as shown by several authors, possibly provides prognostic information concerning biologic behavior. However, the current method for determining PCNA labeling index (LI) is tedious and time consuming like all the nonautomated methods for evaluating cell kinetics, presenting high interobserver and interlaboratory variability and low reproducibility. In the present study, we introduce a semi-automated computer-assisted image analysis method for determining PCNA LI in 38 meningiomas, in parallel with the current nonautomated method. Image analysis technique permits unbiased cell counting, standardizes the degree of staining intensity and provides instant results. By calculating coefficient of variability, the method proved to be highly reproducible. The correlation between the results provided by the nonautomated and the semiautomated image analysis method showed a high agreement between them, with a correlation coefficient, r, of 0.82. In conclusion, we consider that image analysis contributes to the accuracy, reproducibility, and practicality of PCNA LI determination so that along with other useful parameters this significant marker may serve to predict the clinical behavior in meningiomas. PMID- 9228255 TI - P-glycoprotein expression in high grade central osteosarcoma and normal bone cells. An immunohistochemical study. AB - One important mechanism by which multidrug resistance is mediated is the mdr1 gene product, P-glycoprotein (Pgp). Even though chemotherapy, in the treatment of high grade central osteosarcoma (hgc-OS), has led to dramatic improvements in survival rate, a certain percentage of patients still show only a poor response to chemotherapy. To further characterize a potential connection between Pgp and chemotherapy as well as the role of Pgp in tumorigenesis of osteosarcoma, we analyzed Pgp-expression in hcg-OS. Immunohistochemistry was performed on 68 hgc OS samples from 58 patients using the monoclonal antibody JSB-1; in addition, Pgp expression in normal bone cells was studied in 5 human epiphyseal growth plates. 70.5% of all cases stained positive for P-glycoprotein, while 29.5% of the cases were negative. Cases investigated after chemotherapy showed a higher incidence (82.9%) of positive P-glycoprotein immunostaining than cases prior to chemotherapy (64.4%). The Pgp-expression of 34 biopsies was compared with chemotherapy, as determined at the surgical specimen. In these cases, however, no correlation could be established between P-glycoprotein expression of the biopsy and the later response to chemotherapy. 48.4% of the cases with biopsies, initially positive for Pgp, showed a good response in the surgical specimen, while only 27.2% of Pgp-positive biopsies were later classified as non responders. In the normally growing skeleton, positive immunostaining was detected in the area of mineralization of epiphyseal growth plates. Osteoclasts, hypertrophic chondrocytes, and cuboidal osteoblasts showed Pgp-expression, while there was a lack of Pgp in the majority of osteocytes and chondrocytes in the resting and proliferating zone. These data therefore suggest that P-glycoprotein expression in hgc-OS resembles, at least in part, the phenotype of active bone cells. These results may explain why P-glycoprotein, by using immunohistochemistry, in biopsies of osteosarcomas is insufficient to predict the response to chemotherapy. PMID- 9228256 TI - Assessment of proliferative activity, DNA values and some clinicopathologic parameters in mesenchymal tumors. Immunohistochemical and flow cytometric study. AB - Clinicopathologic parameters of 70 consecutive mesenchymal tumors from 63 patients were evaluated. In all these cases, the DNA content was analyzed by flow cytometry, and the expression of proliferative antigen MIB1 and p53 protein was assessed by immunohistochemistry. Our study verified the prognostic usefulness of proliferative indicators, above all MIB1-index, which strongly correlated with tumor grade and independently influenced overall survival. PMID- 9228257 TI - Subacute necrotizing encephalomyelopathy (Leigh's disease): a clinicopathologic study of ten cases. AB - Archival material and clinical data of 10 autopsy cases of Leigh's disease (LS), aged from 44 days to 9 years at death, were reviewed. Development delay, irregular respiration, feeding difficulty, and abnormal eye signs were the most common symptoms. Seizures (five of ten cases) were also frequent. In most patients, the diagnosis of LS was established postmortemly by the presence of symmetrical spongiform lesions affecting several brain centers at autopsy. The histologic examination disclosed associated hypertrophic cardiomyopathy in six cases, while fatty infiltration of the hepatocytes was observed in four cases. Microvesicular degeneration of the renal tubular epithelial cells was also seen in four cases. Our observations suggest that liver and kidney involvement is a component of LS and that this rare entity has to be considered as a polysystematic disorder, able to affect other organs besides the nervous system and the heart, a fact which has not been emphasized enough in the existing literature. PMID- 9228258 TI - Pure heterologous sarcoma of mixed type of the uterine corpus in a postmenopausal patient: a case report. AB - A case of a pure heterologous sarcoma of mixed type (pleomorphic rhabdomyosarcoma and chondrosarcoma) localized in the uterine corpus of a 75-year-old woman is presented. The tumor was investigated by routine morphologic methods, immunohistochemically, and on ultrastructural level. The histogenesis of the tumor and the problems of differential diagnosis with other tumors of the same localization are under discussion. PMID- 9228259 TI - Sarcoid-like reaction in the regional lymph nodes and spleen in gastric carcinoma: a clinicopathologic study of five cases. AB - To clarify the occurrence of sarcoid-like reaction in the spleen of the gastric carcinoma patients, 100 consecutive specimens from gastrosplenectomy were examined. Sarcoid-like reaction was observed in the lymph nodes of 13 cases (13%) and the spleen of five cases (5%). All cases of the latter group were included in the former one. None of them showed any symptoms or signs indicative of systemic sarcoidosis. It seems that the cases with sarcoid-like reaction in the spleen ocurred more frequently in an advanced stage of the gastric cancer than those without this phenomenon. Epithelioid cell granulomas (EPGs) appeared to arise in the periarteriolar lymphoid sheaths of the spleen histologically, but were never found in red pulp or germinal centers. They were composed of groups of epithelioid cells and accompanied by the small lymphocytes and plasma cells. In three cases, scattered eosinophils were also observed among the epithelioid cells. Immunohistochemically, the majority of the intragranulomatous small lymphocytes had T-cell phenotype, while B-cells formed only the minor cellular population. None of the 13 cases contained EPGs in the primary tumor. Our study indicates that sarcoid-like reaction in the spleen is possibly not such a rare phenomenon in the gastric cancer as previously considered and more frequently seen in the advanced stage of the gastric cancer. Sarcoid-like reactions of the regional lymph nodes were more frequently seen in the patients with EPGs in the spleen than in those without. We also suggest that the incidence of sarcoid-like reactions in the spleen is closely related to those in pancreaticosplenic nodes and/or nodes of the hilus of the spleen. PMID- 9228260 TI - Encapsulated uninodular dermatofibrosarcoma protuberans of the dermis: a case report. AB - We report an unusual case of dermatofibrosarcoma protuberans presenting as a small encapsulated uninodular mass, located exclusively in the dermis. Differential diagnostic problems with nodular spindle-shaped cell lesions of the dermis are discussed. A distinct histologic finding was the presence of a myofibroblast-rich fibrous pseudocapsule surrounding the tumor. The possible origin and role of this pseudocapsule are proposed. PMID- 9228261 TI - Malignant peripheral nerve sheath tumor with extensive miliary metastases: a case report. AB - We report the course of a 19-year-old female who presented with a 13-year history of an occipital dermal tumor. Only five weeks after excision, the patient succumbed to cerebral dysregulation due to extensive miliary cerebral, hepatic, and pulmonary metastasis. The histologic features of the surgical specimen and the autopsy material differed suspiciously: a peculiar malignant progression of the tumor seemed to have occurred within five weeks. PMID- 9228262 TI - Hypothalamic osteolipoma: a case report. AB - The authors present a case of an ossified lipoma at tuber cinerum, an incidental finding when performing an autopsy of a 61-year old male. A tight connection to right communicans posterior artery led first to the diagnosis of a calcified saccular aneurysm, but histologic examination revealed an osteolipoma consisting of mature adipose tissue and formation of bone. Intracranial lipomas are neither hamartomas nor true neoplasms, but they are more likely to be congenital malformations. The recent literature is reviewed, and the two most current embryologic concepts of the development of intracranial lipomas are discussed. PMID- 9228263 TI - Small cell carcinoma of the ovary, hypercalcemic type. A case report with immunohistochemical, ultrastructural and cytophotometric analysis and review of the literature. AB - Small cell carcinomas of the ovary, hypercalcemic type, are unilateral tumors occurring almost exclusively in young women. 60% of the cases are associated with hypercalcemia. Microscopic examination shows diffuse clusters of small cells and follicle-like spaces. Immunohistochemically, the tumor cells are positive for epithelial and mesenchymal markers, and ultrastructurally they contain an abundance of dilated rough endoplasmatic reticulum and numerous free ribosomes. By DNA-single cell photometry, the neoplastic cells show a diploid DNA-content. PMID- 9228264 TI - Simultaneous occurrence of a medullary and a papillary thyroid carcinoma in the same patient. AB - We present the case of a patient in whom we diagnosed two different thyroid carcinomas (one on each lobe) of distinct histologic type: one derived from the follicular cells (papillary) and one from the C cells (medullary). They were both diagnosed preoperatively by fine needle aspiration (FNA), and the diagnosis was confirmed with histologic examination. "Inappropriate" staining with neuroendocrine markers was observed in the papillary tumor. Analysis of tumor tissue for the RET oncogene mutations, commonly found in the MEN2 syndromes, was negative. This case supports the view of a common origin for these two tumor types. PMID- 9228265 TI - Sensitivity of delayed gadolinium-enhanced MRI in multiple sclerosis. AB - INTRODUCTION: We performed this study to define the sensitivity of delayed gadolinium-enhanced magnetic resonance imaging (MRI) in detecting active lesions in the brains of patients with multiple sclerosis (MS). MATERIAL AND METHODS: T1 weighted images were obtained in 27 patients with relapsing-remitting or secondary progressive MS before, 5-7 min and 20-30 min after the injection of 0.1 mmol/kg gadolinium-DTPA. RESULTS: One-hundred-and-three enhancing lesions were found on the early and 110 on the delayed scans (increase=6.4%). Six patients had 8 additional lesions in the delayed scans, while 1 patient had 1 more lesion on the early scan. Two of the 12 (17%) patients with no enhancing lesions on the early scans had 2 enhancing lesions on the delayed scans. The average increase of enhancing lesion detection with delayed scanning was 14.5% for those patients who already had enhancing lesions on the early post-contrast scans. A significant increase of the enhancing lesion volume was found with delayed scanning (P=0.004). CONCLUSION: These data indicate that it is possible to increase MRI sensitivity in detecting MS active lesions by delaying the scanning after gadolinium injection. PMID- 9228266 TI - Evaluation of myocardial muscle functional parameters in patients with multiple sclerosis. AB - Structural and functional parameters of myocardium were evaluated with 2D+ Doppler echocardiography in two similar age groups of MS patients: S1: 12 subjects in 3-4 Degrees EDSS (Expanded Disability Status Scale), S2: 12 subjects in 5-7 degrees EDSS. The control group comprised 12 healthy subjects temporarily (at least 1 month) immobilized due to lower limb fractures. Investigations were performed in the supine position and 3 min after tilting to the erect position. Symptoms of organic myocardial injury which might have caused its insufficiency were not observed in any subjects. All structural parameters evaluated by this method did not differ significantly in the examined groups. Symptoms suggesting myocardial insufficiency were found in patients from group S2. Statistically significant decrease of ejection fraction (EF) and cardiac output (CO) was observed in the supine position of S2 patients as compared to S1 and the controls. These symptoms intensified in the erect position in S2 patients and they were accompanied by the decreased values of stroke volume (SV). The fact that in the majority of patients orthostatic hypotonia was not observed and that those disorders were not compensated by significant intensification of heart rate suggest to us that besides disorders resulting from autonomic nervous system dysfunction they may have been caused by secondary myocardial injury in the course of MS. PMID- 9228267 TI - Peripheral neuropathy in acrodermatitis chronica atrophicans - a late Borrelia manifestation. AB - Clinical and/or neurophysiological signs of peripheral neuropathy were found in 64% of 63 consecutive untreated patients with the late borrelial manifestation acrodermatitis chronica atrophicans (ACA). The neuropathy frequency was significantly higher in the patients than in 30 age- and sex-matched control persons of whom 27% had neuropathy findings. The most common neuropathy in ACA was a symmetric distal sensory polyneuropathy. In a subgroup of patients with localized or asymmetric neuropathy, the changes were found more often in extremities with than without visible ACA lesions. Neuropathy symptoms, most often pain and/or paresthesia, were present in 64% of the patients, compared to in 13% of the control persons. Thus, both symptoms and signs of neuropathy were significantly more frequent in patients with untreated ACA than in control subjects. PMID- 9228268 TI - Evoked potential study in facio-scapulo-humeral muscular dystrophy. AB - Nerve conduction velocities (NCVs), somatosensory (SEPs) and auditory evoked potentials (BAEPs) were recorded in 9 patients with facio-scapulo-humeral dystrophy (FSHD) and in 20 age-matched controls. In FSHD patients a significant increase of the nerve distal sensory latencies and of the absolute SEP latencies revealed a subclinical involvement of the afferent sensory pathways, as well as the abnormal slowing of the later components of the BAEPs, pointed to a central auditory dysfunction. Moreover all patients underwent brain MRI that showed the presence of white matter hyperintense lesions in 4 of them (44%). No correlations were found between individual or total number of SEP and BAEP abnormal electrophysiological parameters and severity of WMHL, age, age at onset, duration of the disease or muscular impairment. These findings make the interpretation and pathophysiology of the nervous damage in FSHD rather uncertain. More studies are required to better define the aspects of neurogenic involvement in this type of muscular dystrophy. PMID- 9228269 TI - CADASIL: skin biopsy allows diagnosis in early stages. AB - OBJECTIVES: Our aim was to investigate the diagnostic impact of skin biopsies in CADASIL patients. MATERIALS AND METHODS: Eight consenting CADASIL patients belonging to a German-Caucasian kindred were assessed clinically, genetically, by MRI and skin biopsy. Skin biopsy results were compared to 5 patients suffering from sporadic leucoencephalopathies (control group). RESULTS: Six CADASIL patients presented with symptoms ranging from migraine to severe tetraparesis with dementia. Two clinically unaffected patients had abnormal MRIs. On MRI 7 patients showed various degrees of leucoencephalopathy. One 22-year-old woman with migraine had a normal MRI. Granular, electron dense, osmiophilic material (GEM) was found in skin biopsies of all 8 patients including the 22-year-old woman with migraine and a normal MRI. As shown by genetic linkage analysis she was carrying the disease haplotype. GEM was not found in the control group. CONCLUSION: Our findings substantiate the impact of skin biopsies in defining the carrier status in CADASIL families. PMID- 9228271 TI - Usefulness of short-term video EEG recording with saline induction in pseudoseizures. AB - OBJECTIVES: To study the usefulness of short-term recording of video electroencephalography (VEEG) as an outpatient procedure with placebo induction (PLIN) and intravenous saline in cases of pseudoseizures (Psz). MATERIAL AND METHODS: Fifty cases of suspected Psz were enrolled. They were divided into 2 groups: Group 1 consisted of patients with frank Psz, Group 2 those where diagnosis was uncertain. VEEG recording was done and 10 ml of saline used for placebo-induction. RESULTS: Of 50 patients, 24 (48%) were in Group 1 and 26 (52%) in Group 2. Fifteen (30%) had a spontaneous event during VEEG and 33% had an event only on PLIN. The diagnosis was confirmed in 60 %. In 24% of patients anti epileptic drugs were discontinued. CONCLUSION: Short-term monitoring with VEEG using PLIN is a useful initial screening procedure and in patients where it is inconclusive, long term recordings may be done. PMID- 9228270 TI - Transcranial Doppler echo contrast studies using different colour processing modes. AB - OBJECTIVES: To study the effects of different colour imaging modes on the contrast-medium-enhanced image of the intracranial cerebral arteries. METHODS: Twelve healthy volunteers were studied transcranially after administration of 10 ml BY963 successively with Power Doppler (p-TCCS) and with colour Doppler frequency imaging mode (f-TCCS) in a randomized order. RESULTS: The latency time (mean+/-SD) from the injection until the signal enhancement in the middle cerebral artery was 17.1+/-5.8 s for p-TCCS and 17.8+/-4 s for f-TCCS, and the duration of the optimal diagnostically useful signal enhancement was 44.2+/-8.2 s and 40.2+/-12.6 s respectively. CONCLUSIONS: Based on the measured parameters, both imaging modes were of equal value. Theoretical differences in sensitivity of the two methods play no particular role facing the immense signal enhancement after echo contrast application. PMID- 9228272 TI - Reversible limbic encephalitis caused by ovarian teratoma. AB - A 19-year-old woman developed memory loss followed by psychosis, coma, convulsion, and central hypoventilation requiring mechanical ventilation. MRI of the brain showed minimal changes, and SPECT imaging revealed a small region of increased uptake in the cortex. Intravenous acyclovir and high-dose corticosteroids were administered without any effect. An extensive work-up revealed an elevated serum alpha-fetoprotein (AFP) concentration and the presence of an ovarian tumor. Following resection of the tumor, an immature teratoma by pathology, the patient had significant recovery of her cognitive function with some psychotic sequela. Serum anti-neuronal antibody (anti-Hu) was negative both by immunohistochemistry and by Western blot analysis. A rare combination of paraneoplastic limbic encephalitis and brainstem encephalitis was the suspected diagnosis. Because the tumor contained a neuronal component, we propose an immunologic cross-reaction as the pathomechanism, but the lack of a specific antibody may suggest cell-mediated rather than globulin-mediated immunity. PMID- 9228273 TI - Direct invasion of the brain parenchyma by Mycoplasma pneumoniae. PMID- 9228274 TI - The 26S proteasome: subunits and functions. AB - The 26S proteasome is an eukaryotic ATP-dependent, dumbbell-shaped protease complex with a molecular mass of approximately 2000 kDa. It consists of a central 20S proteasome, functioning as a catalytic machine, and two large V-shaped terminal modules, having possible regulatory roles, composed of multiple subunits of 25-110 kDa attached to the central portion in opposite orientations. The primary structures of all the subunits of mammalian and yeast 20S proteasomes have been determined by recombinant DNA techniques, but structural analyses of the regulatory subunits of the 26S proteasome are still in progress. The regulatory subunits are classified into two subgroups, a subgroup of at least 6 ATPases that constitute a unique multi-gene family encoding homologous polypeptides conserved during evolution and a subgroup of approximately 15 non ATPase subunits, most of which are structurally unrelated to each other. PMID- 9228275 TI - Effects of nucleotides on assembly of the 26S proteasome and degradation of ubiquitin conjugates. AB - We have investigated three aspects of nucleotide usage by the 26S proteasome and its regulatory complex (RC). Both particles hydrolyze the four major ribonucleotides, but ATP and CTP have substantially lower Kms for hydrolysis than do GTP and UTP. The Km for ATP hydrolysis is 15 microm for the 26S proteasome and 30 microm for the regulatory complex. Formation of the 26S proteasome from the RC and the 20S proteasome requires about 5 microm ATP. Although measurable degradation of Ubiquitin(Ub)-lysozyme conjugates occurs in the presence of CTP, GTP, and UTP, the best nucleotide for Ub-conjugate degradation by the 26S proteasome is ATP, with an estimated Km of 12 microm. In summary, our studies show that micromolar concentrations of ATP are sufficient for several 26S proteasome activities. PMID- 9228277 TI - Ubiquitin-mediated degradation of tyrosine aminotransferase (TAT) in vitro and in vivo. AB - Degradation of a protein via the ubiquitin proteolytic pathway involves two successive steps. Covalent attachment of ubiquitin to the target protein and degradation of the tagged substrate by the 26S proteasome. Most native cellular proteins that are targeted by the ubiquitin system are short-lived transcriptional activators and growth and cell cycle regulators, as well as unstable membrane proteins. In the present study we demonstrate the involvement of the system in the degradation of tyrosine aminotransferase (TAT), a key enzyme in intermediary metabolism. In vitro, we have shown that the native enzyme is conjugated and degraded in a system that requires ATP and ubiquitin. Degradation was monitored by following the decrease of catalytic activity as well as disappearance of the protein molecule. The enzyme could be protected from degradation by association with its specific cofactor, pyridoxal phosphate (PLP). In vivo, we prepared cell extracts from livers of animals in which TAT was induced by starvation and corticosteroid administration. The dramatic increase in the level of the enzyme was accompanied by a concomitant increase in the level of specific TAT-ubiquitin adducts. PMID- 9228276 TI - ATPase and ubiquitin-binding proteins of the yeast proteasome. AB - The 26S proteasome is a 2-Megadalton proteolytic complex with over 30 distinct subunits. The 19S particle, a subcomplex of the 26S proteasome, is thought to confer ATP-dependence and ubiquitin-dependence on the proteolytic core particle of the proteasome. Given the complexity of the 19S particle, genetic approaches are likely to play an important role in its analysis. We have initiated biochemical and genetic studies of the 19S particle in Saccharomyces cerevisiae. Here we describe the localization to the proteasome of several ATPases that were previously proposed to be involved in transcription. Independent studies indicate that the mammalian 26S proteasome contains closely related ATPases. We have also found that the multiubiquitin chain binding protein Mcb1, a homolog of the mammalian S5a protein, is a subunit of the yeast proteasome. However, contrary to expectation, MCB1 is not an essential gene in yeast. The mcb1 mutant grows at a nearly wild-type rate, and the breakdown of most ubiquitin-protein conjugates is unaffected in this strain. One substrate, Ub-Proline-beta gal, was found to require MCB1 for its breakdown, but it remains unclear whether Mcb1 serves as a ubiquitin receptor in this process. Our data suggest that the recognition of ubiquitin conjugates by the proteasome is a complex process which must involve proteins other than Mcb1. PMID- 9228278 TI - Accumulation of ubiquitinated proteins in mouse neuronal cells induced by oxidative stress. AB - Ubiquitin protein conjugates are commonly detected in neuronal brain inclusions of patients with neurodegenerative disorders. The failure to eliminate the ubiquitin-protein deposits in the degenerating neurons may result from changes in the activity of the ubiquitin/ATP-dependent proteolytic pathway. This proteolytic pathway plays a major role in the degradation of short lived, abnormal and denatured proteins. Cadmium is a potent cell poison and is known to affect the ubiquitin pathway and to cause oxidative stress. Increases in protein mixed disulfides (Pr-SSG) and decreases in glutathione (GSH) are often used as markers of oxidative stress. To investigate the relationship between the ubiquitin pathway and cellular glutathione (GSH), we treated HT4 cells (a mouse neuronal cell line) and rat mesencephalic primary cultures with different concentrations of the heavy metal. We observed marked increases in Pr-SSG as well as decreases in GSH, after exposure of HT4 cells or primary mesencephalic cultures to Cd2+. Furthermore, our results show that Cd2+ induced the accumulation of ubiquitinated proteins. Detection was by Western blotting of total cell extracts probed with antibodies that recognize ubiquitin-protein conjugates. These results suggest that the ubiquitin-pathway is closely involved in the cell response to cadmium mediated oxidative stress. PMID- 9228279 TI - The 26S-proteasome: regulation and substrate recognition. AB - There is extensive reprogramming of the ATPase regulators of the 26S proteasome before the programmed elimination of the abdominal intersegmental muscles (ISM) after eclosion in Manduca sexta [1]. This extensive ATPase reprogramming only occurs in ISM which are destined to die and not in flight muscle (FM). The MS73 ATPase also increases in the proleg retractor muscles which die at a developmentally different stage to ISM. The non-ATPase regulator S5a shows a similar increase to the ATPase regulators. We have cloned the Manduca SUG2 ATPase and shown that this ATPase is a component of the 26S proteasome. This ATPase shows a similar increase in concentration to the other ATPases in 26S proteasomes before muscle death. The SUG2 ATPase is also associated with other smaller complexes besides the 26S proteasome which act as activators of the 26S proteasome. Finally, in a yeast two-hybrid genetic screen we have identified a protein in human brain which interacts with the MS73 ATPase (and human S6). The interacting protein contains 6 ankyrin repeats and is co-immunoprecipitated with anti-MS73 antiserum after in vitro transcription/translation. The ankyrin repeat protein may interact with the MS73 ATPase as part of the substrate recognition process by the 26S proteasome. Many proteins degraded by the 26S proteasome contain ankyrin repeats, e.g. IkB and some cyclins: binding through ankyrin repeats to an ATPase regulator may complement protein ubiquitination and S5a binding as recognition signals by the 26S proteasome. PMID- 9228280 TI - Proteasome inactivation upon aging and on oxidation-effect of HSP 90. AB - Increases of oxidatively modified protein in the cell have been associated with the aging process. Such an accumulation of damaged protein may be the result of increase in the rate of protein oxidation and/or decrease in the rate of degradation of oxidized protein. The multicatalytic proteinase or proteasome is known to be the major proteolytic system involved in the removal of oxidized protein. We have reported that, after isolation of the 20S proteasome from the liver of young and old male Fischer 344 rat, out of the three peptidase activities (chymotrypsin-like, trypsin-like and peptidyl-glutamyl peptide hydrolase) we assayed with fluorogenic peptides, the peptidyl-glutamyl peptide hydrolase activity was declining with age to a value approximately 50% of that observed for protease purified from young rats. The proteasome was subjected to metal catalyzed oxidation to determine the susceptibility of the different peptidase activities to oxidative inactivation. Both trypsin-like and peptidyl glutamyl peptide hydrolase activities were found sensitive to oxidation. Treatment of the proteasome with 4-hydroxy-2-nonenal, a major lipid peroxidation product, was also found to inactivate the trypsin-like activity. However, the trypsin-like activity was protected from inactivation by metal catalyzed oxidation in proteasome preparations contaminated with HSP 90, a protein that often copurifies with the proteasome. Upon addition of HSP 90 to pure 20S active proteasome, the trypsin-like activity was protected from inactivation by metal catalyzed oxidation and from inactivation by treatment with 4-hydroxy-2-nonenal. These results suggest a possible intervention of HSP 90 in response to oxidative stress in preventing the inactivation of the proteasome by oxidative damage. PMID- 9228281 TI - Complex mechanisms for c-fos and c-jun degradation. AB - c-fos and c-jun proto-oncogenes have originally been found in mutated forms in murine and avian oncogenic retroviruses. They both define multigenic families of transcription factors. Both c-jun and c-fos proteins are metabolically unstable. In vivo and in vitro work by various groups suggests that multiple proteolytic machineries, including the lysosomes, the proteasome and the ubiquitous calpains, may participate in the destruction of c-fos and c-jun. The relative contribution of each pathway is far from being known and it cannot be excluded that it varies according to the cell context and/or the physiological conditions. It has been demonstrated that, in certain occurrences, the degradation of both c-fos and c jun by the proteasome in vivo involves the ubiquitin pathway. However, the possibility that proteasomal degradation can also occur in a manner independent of the E1 enzyme of the ubiquitin cycle remains an open issue. PMID- 9228282 TI - Prosomes (proteasomes) changes during differentiation are related to the type of inducer. AB - The core of the 26S proteasome, the 20S prosome, is a highly organized multi protein complex found in large amount in malignant cells. Differentiation of several cell lines, including the monoblastic U937 and the lymphoblastoid CCRF CEM, is accompanied by a general decrease in the prosome concentration when phorbol-myrirtic-acetate (PMA) and retinoic acid plus dihydroxyvitamine D3 (RA+VD) are used. Incubation of U937 cells for three days with PMA or RA+VD causes differentiation, but the resulting patterns of prosome labeling in the cell and on the plasma membrane are not the same. In contrast, the same kind of prosome changes occur in U937 and CCRF-CEM cells when PMA is used as inducer. The intracellular distribution of prosomes is also linked to malignancy and differentiation. Prosomes are found in the nucleus and the cytoplasm of cancer cells; and treatment with RA+VD decreases the prosomes in the nucleus whereas PMA causes various prosome proteins changes. These results indicate that prosomes are important in cell regulation and in the expression of malignancy. PMID- 9228283 TI - Antigen processing by proteasomes: insights into the molecular basis of crypticity. AB - Eight to eleven amino acid residues are the sizes of predominant peptides found to be associated with MHC class I molecules. Proteasomes have been implicated in antigen processing and generation of such peptides. Advanced methodologies in peptide elution together with sequence determination have led to the characterisation of MHC class I binding motifs. More recently, screening of random peptide phage display libraries and synthetic combinatorial peptide libraries have also been successfully used. This has led to the development and use of predictive algorithms to screen antigens for potential CTL epitopes. Not all predicted epitopes will be generated in vivo and the emerging picture suggests differential presentation of predicted CTL epitopes ranging from cryptic to immunodominant. The scope of this review is to discuss antigen processing by proteasomes, and to put forward a hypothesis that the molecular basis of immunogenicity can be a function of proteasomal processing. This may explain how pathogens and tumours are able to escape immunosurveillance by altering sequences required by proteasomes for epitope generation. PMID- 9228284 TI - Protein degradation by the proteasome and dissection of its in vivo importance with synthetic inhibitors. PMID- 9228285 TI - Proteasome and myogenesis. AB - In this report, we examine the involvement of the ubiquitin-proteasome pathway during fusion and differentiation of myoblast primary cell cultures. Up regulation of proteasome was observed at the maximum fusion rate and was preceded by an increase of unidentified ubiquitin-conjugates. Cell permeable proteasome inhibitors prevent fusion as do antisense oligodesoxyribonucleotides targetted to three proteasome subunits. Identical results were obtained using E3 ubiquitin ligases dipeptide inhibitor. Involvement of the ubiquitin-proteasome pathway in the regulation of myogenic factors was hypothesized. PMID- 9228286 TI - The 26S proteasome: a dynamic structure. AB - The proteasomal system consists of a proteolytic core, the 20S proteasome, which associates in ATP-dependent and independent reactions with endogenous regulators providing specific substrate binding sites, chaperone function and regulation of activity to the protease. The best known regulators of the 20S proteasome are the 11S and the 19S complexes. Three subunits of the 20S proteasome and the two subunits of the 11S regulator are induced by gamma-Interferon. However, there are no indications for an influence of gamma-interferon on the subunit composition of the 19S regulator and only a few data exist about the dynamics of this complex. The analysis of 19S regulator subunits from yeast mutants reveals that the ATPases appear to be stringently organized in the 26S complex, while peripheral non-ATPases, such as S5a, might serve as subunits which shuttle substrates to the enzyme. A novel non-ATPase has been cloned, sequenced and identified in a complex besides the 19S regulator, the function of which is presently unknown. The dynamic structure of the 26S proteasome is also characterized by transient associations with components such as the modulator and isopeptidases. Certain viral proteins can also be associated with components of the proteasomal system and alter enzymatic activities. PMID- 9228287 TI - Structural and functional properties of proteasome activator PA28. AB - The proteasome activator PA28 or 11S regulator is a protein complex composed of two different but homologous polypeptides, termed PA28alpha and PA28beta. The purified activator protein (approximately 200 kDa) is a ring-shaped heteromultimer containing the two polypeptides, possibly with an (alpha3beta3 stoichiometry. The activator, which by itself shows no hydrolytic activity elicits activation of the proteasome's multiple peptidase activities by binding to the terminal rings of the proteinase. In vitro, active PA28 can be reconstituted from isolated alpha and beta subunits, yielding two different oligomers: with the single alpha subunit, PA28alpha homomultimers with moderate stimulatory activity toward 20S proteasomes are obtained whereas isolated beta subunits are unable to form oligomers and are devoid of stimulatory activity. However, in the presence of both subunits, alphabeta heteromultimers form, concomitant with restoration of full stimulatory activity. The recent finding that PA28 modulates the proteasome-catalyzed production of antigenic peptides presented to the immune system on MHC class I molecules indicates a cellular function of the activator in antigen processing. PMID- 9228288 TI - Expression of subunits of the 19S complex and of the PA28 activator in rat skeletal muscle. AB - A precise knowledge of the role of subunits of the 19S complex and the PA28 regulator, which associate with the 20S proteasome and regulate its peptidase activities, may contribute to design new therapeutic approaches for preventing muscle wasting in human diseases. The proteasome is mainly responsible for the muscle wasting of tumor-bearing and unweighted rats. The expression of some ATPase (MSS1, P45) and non ATPase (P112-L, P31) subunits of the 19S complex, and of the two subunits of the PA28 regulator, was studied in such atrophying muscles. The mRNA levels for all studied subunits increased in unweighted rats, and analysis of MSS1 mRNA distribution profile in polyribosomes showed that this subunit entered active translation. By contrast, only the mRNA levels for MSS1 increased in the muscles from cancer rats. Thus, gene expression of the proteasome regulatory subunits depends on a given catabolic state. Torbafylline, a xanthine derivative which inhibits tumor necrosis factor production, prevented the activation of protein breakdown and the increased expression of 20S proteasome subunits in cancer rats, without reducing the elevated MSS1 mRNA levels. Thus, the increased expression of MSS1 is regulated independently of 20S proteasome subunits, and did not result in accelerated proteolysis. PMID- 9228289 TI - Regulation of proteasome structure and function. AB - Proteasomes are cylindrical particles made up of a stack of four heptameric rings. In animal cells the outer rings are made up of 7 different types of alpha subunits and the inner rings are composed of 7 out of 10 possible different beta subunits. Regulatory complexes can bind to the ends of the cylinder. We have investigated aspects of the assembly, activity and subunit composition of core proteasome particles and 26S proteasomes, the localization of proteasome subpopulations, and the possible role of phosphorylation in determining proteasome localization, activities and association with regulatory components. PMID- 9228290 TI - Structure and structure formation of the 20S proteasome. AB - Eukaryotic 20S proteasomes are complex oligomeric proteins. The maturation process of the 14 different alpha- and beta-subunits has to occur in a highly coordinate manner. In addition beta-subunits are synthesized as proproteins and correct processing has to be guaranteed during complex maturation. The structure formation can be subdivided in different phases. The knowledge of the individual phases is summarized in this publication. As a first step the newly synthesized monomers have to adopt the correct tertiary structure, a process that might be supported in the case of the beta-subunits by the intramolecular chaperone activity postulated for the prosequences. Subsequently the alpha-subunits form ring-like structures thereby providing docking sites for the different beta subunits. The result most likely is a double ring structure (13S precursor) representing half-proteasomes, which contain immature proproteins. Two 13S precursors associate to form the proteolytically inactive 16S assembly intermediate which still contains unprocessed beta-monomers. In addition the chaperone Hsc73 is present within these particles suggesting an essential role during the structure formation process. The processing of monomers with an N terminal threonine occurs within the 16S particles and is achieved autocatalytically by two subsequent processing events finally leading to the mature, active 20S proteasome. PMID- 9228291 TI - Proteasome (prosome) associated endonuclease activity. AB - The 20S proteasome (prosome) is a highly organized multiprotein complex with approximate molecular weight of about 700 kDa. Whilst the role of the proteasome in the processing and turnover of cellular proteins is becoming clearer, its relationship with RNA remains still obscure. Here we focus on the nature and function of proteasome associated endonuclease activity. Thus the involvement of a proteasome alpha-type subunit in RNA-degradation, the catalytic requirements, the interaction of proteasomes with their RNA-substrate and the identification of a well defined cleavage site in the 3'UTR of short-lived cellular mRNAs will be described in detail. All data indicate that proteasomes associated endonuclease activity could be involved in post-transcriptional gene control at the level of translation. PMID- 9228292 TI - Synthetic peptide-based activators of the proteasome. AB - The development of small molecule peptide-based activators of the 20S proteasome or multicatalytic proteinase complex was initiated. The enhancement of antigen presentation by transfection of the protein activator PA28alpha into a mouse fibroblast cell line [10] supports the potential use of small molecule activators in stimulating the immune response. Four classes of peptide-based activators were synthesized, i.e. peptidyl alcohols, esters, p-nitroanilides and nitriles. These compounds markedly and reversibly stimulated the hydrolysis of suc-LLVY-MCA, Z LLE-NA and Z-GPALG-p-aminobenzoate as well as hydrolysis of the decapeptide angiotensin I. Stimulation was due to a decrease in the Km and increase in the Vmax of the substrate. In general, the EC50 for activation ranged from 50-150 mM and maximal stimulation varied from 3 to 15 fold depending on the activity measured. Z-IE(Ot-Bu)AL-p-nitroanilide, a proteasome substrate, markedly stimulated the hydrolysis of Z-GPALG-pAB by binding to a saturable high affinity site distinct from its binding site as substrate. Since all effective activators contain hydrophobic groups in positions P1-P5, low aqueous solubility is a limitation of these compounds. Competition experiments suggest that these activators bind to the same site as PA28. PMID- 9228294 TI - Biochemical properties of insect and crustacean proteasomes. PMID- 9228293 TI - Eubacterial proteasomes. AB - Proteasomes are large, multisubunit proteases with highly conserved structures. The 26S proteasome of eukaryotes is an ATP-dependent enzyme of about 2 MDa, which acts as the central protease of the ubiquitin-dependent pathway of protein degradation. The core of the 26S complex is formed by the 20S proteasome, an ATP independent, barrel-shaped protease of about 700 kDa, which has also been detected in archaebacteria and, more recently, in eubacteria. Currently, the distribution of 20S proteasomes in eubacteria appears limited to the actinomycetes, while most other eubacteria contain a related complex of simpler structure. PMID- 9228296 TI - Maternal response to paternal trophoblast antigens. AB - PROBLEM: What is the function of the immunoglobulin (Ig) G antibody bound to trophoblast in normal pregnancy, and what is the antigen? METHOD: IgG was acid eluted from term human placental microvesicles and reacted with the antigen, R80K, left on the vesicles. The eluted antibody was used to detect the antigen on monocytes, lymphocytes, and lymphoblastoid cell lines. The eluted antibody is highly polymorphic, but monoclonal antibodies (mAbs) were made against conserved regions of the molecule. These also reacted with the murine equivalent of the human R80K and were used in inhibition studies of natural killer (NK) cell killing and the mouse abortion models, CBA x DBA2 F1 resorption in CBA females, the endotoxin-induced resorption model, and a sonic stress-induced murine resorption model. RESULTS: All 600 syncytiotrophoblast microvesicle preparations of human term placenta had IgG antibody bound, elutable at pH 3.0. The eluted antibody reacted with about 15% of unrelated human placentae. In horses mares make detectable antibody early in pregnancy, at about the time of implantation. The IgG antibody was bound to an 80-kDa protein (R80K) also detected on B lymphocytes and monocytes. In HLA homozygous lymphoblastoid B cell lines, which reacted with one or more eluted antibodies, had a pattern of cytotoxicity independent of HLA Class I; and as a single 80-kDa peptide chain, R80K did not resemble HLA molecules. Genetic studies in horses show that of the two paternal allotypes of R80K detectable by placental alloantibodies, only one, usually the grandpaternal one, is present in all the placentae of a sibship. Two of 26 eluted human antibodies had affinity for K562 and inhibited killing by human peripheral blood NK cells. One mAb, BA11, against a conserved site on R80K inhibited killing of K562, and also reacted with the murine R80K homologue. BA11 inhibited murine NK cell killing and virtually completely inhibited three NK cell-dependent mouse resorption models. CONCLUSION: R80K protein is a target molecule for NK cell activity expressed on all placentae. It has a polymorphic alloantigenic determinant completely covered with maternal antibody in all successful term pregnancies. In murine NK cell-dependent models of abortion, a mAb against a monomorphic determinant present in human and murine R80K prevents abortion very effectively. It seems that the R80K molecule must be covered with antibody to prevent NK attacks on trophoblast. PMID- 9228295 TI - Interferon tau: a novel pregnancy recognition signal. AB - PROBLEM: Trophectoderm of ruminant conceptuses (embryo and associated membranes) secretes tau interferons (IFNtau) as the pregnancy recognition signal. How does it act? METHOD: Review of current data. RESULTS: IFNtau acts on uterine epithelium to suppress transcription of the genes for estrogen receptor and oxytocin receptor. This blocks development of the uterine luteolytic mechanism and, therefore, release of luteolytic pulses of prostaglandin F2alpha, but it has no effect on expression of the progesterone receptor. Maintenance of progesterone secretion by the corpus luteum ensures establishment and maintenance of pregnancy. Secretion of IFNtau on days 12-15 for sheep and days 14-17 for cows and goats is essential for pregnancy recognition. CONCLUSION: We propose that IFNtau affects endometrial gene expression by activating the Jak/Stat pathway, which results in formation of the ISGF3alpha transcription factor complex. ISGF3alpha binds to interferon-stimulated response elements and activates transcription of interferon-responsive genes such as interferon regulatory factor 1 (IRF-1) which, in turn, activates expression of the negative-acting transcription factor IRF-2. Pregnancy (or intrauterine injection of roIFNtau) results in a transient increase in endometrial IRF-1 expression followed 36-48 hr later by a sustained increase in IRF-2. We propose that IRF-2, or an IFNtau induced negative regulatory factor like IRF-2, suppresses expression of the estrogen receptor gene and directly or indirectly blocks expression of the gene for oxytocin receptor to abrogate the uterine luteolytic mechanism and ensure the establishment of pregnancy. PMID- 9228298 TI - Interferon-gamma production by the human preimplantation embryo. AB - PROBLEM: This study demonstrated that the human embryo produces interferon-gamma (IFNgamma). It is important to know whether IFNgamma can be produced before implantation. Therefore the aim of this study was to evaluate the profile of IFNgamma production between days 2 and 5 after in vitro fertilization. METHOD: Twenty embryos were cultured from day 2 to 5 after fertilization. The embryo stages were checked each day and the media refreshed. IFNgamma levels were estimated by an enzyme-linked immunosorbent assay. RESULTS: All embryos produced measurable IFNgamma at least for 1 day. Yields of IFNgamma were: 0.46 +/- 0.45 (n = 4) on day 2, 0.69 +/- 0.52 (n = 19) on day 3, 0.73 +/- 0.52 (n = 15) on day 4, 0.55 +/- 0.32 (n = 11) IU/ml on day 5, respectively. There was no significant difference in the IFNgamma production between in vitro culture days or between the developmental stages of embryos. CONCLUSION: IFNgamma is produced by all the embryos and seems to peak between days 3 and 4, which is just before implantation. PMID- 9228297 TI - Immune suppression and Th1/Th2 balance in pregnancy revisited: a (very) personal tribute to Tom Wegmann. AB - PROBLEM: The paradigm of local suppression necessary to understand the survival of the fetal allograft is often compared with the host-tumor relationship. METHODS: We investigated two components of local immune suppression: placenta induced immunosuppression, which is mediated at least in part by a soluble factor of low molecular weight that can induce anergy in lymphocytes, and interleukin-10 (IL-10). RESULTS: We show that enhancement of IL-10 production in the decidua and placenta after alloimmunization requires the presence of Asialo GM1+ cells. Placenta-induced immunosuppression is linked with defects in phosphorylation of some components of the T cell receptor. CONCLUSION: NK cells could be in fact regulatory cells pushing maternal immune response toward a Th2 profile, beneficial for fetal survival, or toward a Th1 type of immune response, which acts in synergy. Modulation of TcR may represent a new mechanism for maternal fetal tolerance. PMID- 9228299 TI - Cytokine-leukocyte networks and the establishment of pregnancy. AB - PROBLEM: Factors in seminal plasma stimulate an intense but transient inflammatory response in the murine endometrium at mating. The aim of our current studies is to delineate the cytokine-leukocyte interactions comprising this response and to elucidate the significance of these events in changes in the maternal immune system and as determinants of pregnancy outcome. METHOD: We have reviewed our recent findings. RESULTS: Transforming growth factor (TGF)-beta1 has been identified as the inflammation-inducing moiety in seminal plasma. Seminal TGFbeta1 initiates endometrial leukocyte infiltration by up-regulating epithelial cell expression of granulocyte-macrophage colony-stimulating factor. Other cytokines and chemokines including regulated and normal T-cell expressed and secreted (RANTES), macrophage inflammatory protein (MIP)-1alpha, MIP-1beta, and monocyte chemotactic protein-1 are also implicated as mediators of macrophage and granulocyte recruitment and activation. One consequence of this inflammatory response is the induction of a transient state of hyporesponsiveness to paternal major histocompatibility class I antigens. CONCLUSION: Our studies suggest that semen may play a critical role in providing the antigenic and environmental signals necessary to initiate an appropriate maternal immune response to the conceptus during pregnancy. PMID- 9228300 TI - Distribution of type-I interferon-receptors in human first trimester and term placental tissues and on isolated trophoblast cells. AB - PROBLEM: Type-I interferon (IFN) is the protein recognizing pregnancy in ruminants. Although IFN is secreted in early pregnancy, its role is not still clear in other species. Like other cytokines, IFN exerts its biological functions through specific membrane receptors. We have investigated the potential action of IFN in human pregnancy by studying the distribution of the receptors in the human placenta. METHOD: Reactivity to monoclonal antibodies (mAbs) to the type-I IFN receptor (R) was analyzed by immunohistochemistry in human placental tissues and in cytospins of first trimester trophoblast cells. RESULTS: Type-I IFN-R immunoreactivity was observed mostly in first trimester villous cytotrophoblasts and in the cytotrophoblast cell columns. Trophoblast in the decidua, the epithelium of the uterine glands, and most of the isolated trophoblast cells were also immunoreactive. CONCLUSION: The expression of type-I IFN-R in the highly proliferating and migrating trophoblast suggests that this cytokine has a role in trophoblast growth and invasion. PMID- 9228301 TI - Histochemical evaluation of placental dipeptidyl peptidase IV (CD26) in pre eclampsia: enzyme activity in villous trophoblast indicates an enhanced likelihood of gestational hypertensive disorders. AB - PROBLEM: To determine whether differences in placental dipeptidyl peptidase IV (DPP IV, CD26) activities occurred in hypertensive complicated pregnancies as compared with uncomplicated pregnancies. METHOD: DPP IV activity was detected with H-Gly-Pro-4M2NA as the substrate in placental cryostat sections from 65 patients with gestational hypertension and 67 patients with uncomplicated pregnancies. The graduated intensities of the reaction product in the villous trophoblast were scored semiquantitatively by light microscopy and were related to the relative frequencies of hypertensive disorders (proportional odds model). After detection of enzyme activity, the same tissue samples were homogenized and used for kinetic fluorometric measurements. RESULTS: Enhanced villous trophoblastic DPP IV activity was significantly associated with an increased frequency of proteinuric hypertension in pregnant women (cumulative odds ratio theta1 = 1.6; P < 0.01). CONCLUSION: This study demonstrates for the first time that increased villous trophoblastic DPP IV activity indicates an increased likelihood of the presence and of the severity of hypertensive disorders in pregnancy. PMID- 9228302 TI - Uterine NK cells and trophoblast HLA class I molecules. AB - PROBLEM: To investigate the proposal that NK cells in decidua may control trophoblast migration during implantation of the human placenta. METHOD: Use Mab specific for HLA-G and for HLA-C in association with flow cytometry and immunoprecipitation to determine the expression of these HLA molecules by trophoblast. Expression of Killer inhibitory/activatory receptors (KIR/KAR) and the CD94 receptor by decidual NK cells was also studied. RESULTS: Extravillous trophoblast expressed HLA-G and HLA-C in both beta2m-associated form and as free heavy chains. KIR and KAR are expressed by decidual NK cells. The repertoire of receptors varied between different women and also between blood and decidual NK cells from the same women. The expression of CD94 was also different between blood and decidual NK cells. CONCLUSION: The recognition of HLA-G/HLA-C by KIR/KAR and CD94 could provide a mechanism by which decidual NK cells control trophoblast migration. PMID- 9228303 TI - Uterine natural killer cells do not require interleukin-2 for their differentiation or maturation. AB - PROBLEM: Natural Killer lymphocytes (NK cells) from the pregnant uterus and from other tissues in pregnant and nonpregnant mammals can be stimulated by interleukin-2 (IL-2) during culture to become Lymphokine Activated Killer (LAK) cells. The susceptibility of cultured trophoblast cells to lysis by LAK cells raises the enigma of why uterine (u) NK cells that are characterized by morphology and by surface phenotyping as "activated," and thus potentially damaging to the placenta, become localized to implantation sites during normal rodent gestation. METHOD: uNK cells migrating from explant cultures of the metrial gland were assessed for expression (mRNA and protein) of each chain of the IL-2 receptor (IL-2R). Implantation sites from transgenic mice lacking a functional IL-2 gene were examined histologically for the differentiation of mature, granulated uNK cells. RESULTS AND CONCLUSION: Early post-implantation, mRNA from migrating uNK cells contains transcripts for all three chains of the IL 2R. Only IL-2Rgamma was expressed at day 12 of gestation; expression of this gene was also lost by day 16. Loss of IL-2R transcription did not result in loss of protein expression; however, it did coincide with loss of uNK cell viability in vivo. Apparently normal differentiation of uNK cells occurred in IL-2(-/-) mice and in doubly mutant IL-2(-/-).beta2m(-/-) mice. Thus, despite uNK cell expression of the full IL-2R at day 8 of gestation, IL-2 is not required for the maturation of uNK cells to their fully granulated form or for normal placental development. PMID- 9228304 TI - Decidual infiltration and activation of macrophages leads to early embryo loss. AB - PROBLEM: There is considerable controversy concerning the root cause and mechanisms of early embryo loss. It has been suggested that most pregnancy losses occur due to morphogenetic anomalies of the embryo. It has also been suggested that the maternal specific immune system rejects the embryo. METHODS: Existing data on the cell and molecular biology of early embryo loss in murine experimental models is reviewed. RESULTS: Using the CBA(female) x DBA/2(male) model of early embryo loss, it has been established that maternal inflammatory cells infiltrate the decidua basalis of all implantation sites within 48 hr after implantation. For most embryos, the relatively low numbers of macrophages (Mphi) and natural killer-like (NK-like) cells of maternal origin remain relatively constant after day 8, whereas 20-30% of the embryos show a significant increase in inflammatory cells in the maternal decidua, corresponding to the incidence of early embryo resorption visible at day 12. Evidence will be reviewed to suggest that decidual NK-like cells are not cytolytic but may be producing the Mphi activating cytokine interferon gamma (IFN-gamma), which activates decidual Mphi and other cells. Furthermore, embryo loss is ameliorated by in vivo treatment with anti-IFNgamma or anti-NK antisera, indicating that NK-like cells and/or IFNgamma are required for embryo loss, but not for embryo survival. In resorbing embryos, the inflammatory Mphi show evidence of having been primed during early pregnancy, in that in vitro incubation with lipopolysaccharide induced the production of tumor necrosis factor alpha and nitric oxide. CONCLUSION: These findings support the concept that early embryo loss is a nonspecific event mediated by the triggering of cytotoxin production by primed decidual macrophages. PMID- 9228305 TI - Maternal anti-placental cell-mediated reactivity and spontaneous abortions. AB - PROBLEM: Does Th1/Th2 balance determine pregnancy outcome, and if so, what determines Th1/Th2 balance in pregnancy? METHOD: Review and synthesis of existing data. RESULTS: A bias toward Th1 is strongly correlated with pregnancy failure in mice and humans. Pregnancy usually shifts the balance toward Th2 and placental factors/progesterone and progesterone-stimulated CD8+ T cell production of suppressor factor; TGFbeta2 and IL4/10 may be responsible. The bias toward a Th1 response may result from intracellular parasitic infection and other as yet undefined factors. CONCLUSION: The Th1/Th2 balance thesis appears to be valid. PMID- 9228306 TI - Induced termination of pregnancy by purified extracts of Azadirachta Indica (Neem): mechanisms involved. AB - PROBLEM: To develop a self-administered, orally delivered method for abrogation of early pregnancy. METHOD: Use of purified Neem extracts containing immunomodulators stimulating Th1 cells and macrophages; test animals, rats, baboons, and monkeys, onset of pregnancy confirmed by surgery and counting of implants on day 7 in rats and by chorionic gonadotropin (CG) and progesterone assays in primates; termination defined by complete resorption on day 15 in rats and by bleeding and decline of CG and progesterone in baboons. RESULTS: Pregnancy was terminated successfully in both rodents and primates with no significant side effects. Fertility was regained in both species after one or two irregular cycles. Progeny born had normal developmental landmarks and mothered normal litters in the course of time. The active principle in Neem has been partially fractionated by activity-guided purification. A cascade of events are involved in abrogation of pregnancy. In primates, a decrease in progesterone is an early event. A transient increase in CD4 and CD8 cells is noted in spleen at 96 hr and in mostly CD8 cells in mesenteric lymph nodes. Treatment causes an elevation of both immunoreactive and bioactive TNF-alpha and gamma-interferon in serum, mesenteric lymph nodes, and foetoplacental tissue. CONCLUSION: Immunomodulators of plant origin are potentially usable for termination of unwanted pregnancy PMID- 9228307 TI - Murine T cell determination of pregnancy outcome: I. Effects of strain, alphabeta T cell receptor, gammadelta T cell receptor, and gammadelta T cell subsets. AB - PROBLEM: T cells bearing alphabeta T cell receptor (TcR) and gammadelta TcR are present at the fetomaternal interface, and the latter, which express surface activation markers, can react with fetal trophoblast cell antigens. What is the role of these cells? METHOD: Using stress-abortion-prone DBA/2-mated CBA/J and abortion-resistant C57/B16 mice, alphabeta, gammadelta, and CD8+/- T cell subsets were measured in spleen and uterine decidua. The effect of immunization against abortion and administration of anti-TcR antibody in vivo was examined. Cytokine synthesis was measured by intracellular staining of Brefeldin A-treated cells. RESULTS: Abortion-prone matings showed an unexpected accumulation of gammadelta T cells beginning in the peri-implantation period and this was suppressed by immunization against abortion. The immunization deleted gammadelta T cells producing the abortogenic cytokines, TNF-alpha and gamma-interferon, and increased production of the anti-abortive cytokines, IL-10 and transforming growth factor-beta2 (TGF-beta2). Immunization also boosted the number of alphabeta T cells which were present in the decidua as early as 2 days after implantation. In vivo injection of GL4 (anti-delta) depleted gammadelta T cells producing Th1 cytokines in the peri-implantation period, and prevented abortions, whereas H57 (anti-beta) decreased the number of alphabeta T cells and led to 100% abortions. CD8+ T cells present in peri-implant decidua before onset of abortions were mostly alphabeta TcR+, although some were gammadelta+. Changes in gammadelta and alphabeta T cells in pregnancy were most dramatic in uterine tissue. CONCLUSION: Although decidual gammadelta T cells after formation of a distinct placenta and fetus produce anti-abortive TGF-beta2-like molecules and IL-10, prior events can lead to abortion. High local production of TNF-alpha and gamma interferon develop during the peri-implantation phase because of an excessive increase in the Th1 cytokine+ subset of gammadelta cells; these cytokines may be contributed by other tissues in decidua, and the contribution of bioactive factors by gammadelta T cells may augment the cytokine pool. In contrast, alphabeta T cells (which may be inactivated by stress that causes abortions) may mediate the anti-abortive effect of alloimmunization. Alloimmunization involves a shift from a Th1 to a Th2 pattern in the gammadelta T cells in decidua. PMID- 9228308 TI - Recent studies on the structure of the grc region in the rat. AB - PROBLEM: To explore the nature of the genes in the grc. METHOD: Chromosome walking of the grc and sequencing new genes. RESULTS: The RT1.O,N genes have been aligned, and two new types of genes, RT1.S1,2 and Rprl,2,3, have been discovered and mapped. An extensive physical map of the grc region is presented. CONCLUSION: The disease associations in both the rat and the human appear to cluster in two regions that map in approximately the same place in both species, given the translocation that occurred in the major histocompatibility complex (MHC) of the rat relative to the MHC of the human. PMID- 9228309 TI - Isolated partial rupture of the anterior cruciate ligament. PMID- 9228310 TI - Isolated partial rupture of the anterior cruciate ligament. Long-term follow-up of 56 cases. AB - The majority of previous studies on partial ruptures of the anterior cruciate ligament (ACL) include a relatively large proportion of knees with associated intra-articular injury or collateral ligament tear that contributes to an increase in the symptoms of instability and further deterioration of knee function. In the present study only patients with isolated, partial ruptures of the ACL were evaluated. Fifty-six patients with one injured knee were examined after a median of 5.3 (range 2.0-12.7) years using the IKDC evaluation form, Lysholm knee function score and Tegner activity score. Of the 56 knees, 6 underwent autologous reconstruction due to early progression to complete rupture. Of 34 knees evaluated for laxity, 25 had a negative Lachman test and 7 a positive (+) Lachman. In 2 knees a Lachman ++ result and a positive pivot shift were found. With instrumented laxity testing 24 knees had 2 mm or less difference in laxity compared with the contralateral uninjured knee. The largest side-to-side difference in knee laxity was 4.5 mm. Lysholm score was median 86 (range 52-100) points, and 62% had good or excellent knee function. A significant decline in activity was seen. Only 10 patients (30%) resumed their preinjury activities. We find that the majority of patients with an isolated, partial rupture of the ACL have an acceptable knee function and a stable knee after a median 5 years follow up. There is, however, a marked reduction in activity. PMID- 9228311 TI - T-Fix anchor sutures for arthroscopic meniscal repair. AB - The results of arthroscopically repaired meniscal tears with the T-Fix system in a short-term follow-up of 6 months was assessed in a non-comparative, prospective study. The T-Fix device consists of a short, rigid Delrin T attached to a braided, non-absorbable, polyester suture which is preloaded inside and deployed through a delivery (spinal) needle. The T grabs inside the tissue and provides an anchor for the suture. Twenty menisci in 20 patients (mean age 29 years) were repaired. Sports-related injuries were documented in 18 patients. In 15 patients, meniscus tears were repaired 6 months or more after injury. Half of the patients had isolated meniscus injuries. Associated injuries included anterior cruciate ligament (ACL), medial or lateral collateral ligament ruptures. These were not treated at the time of meniscal surgery except for an ACL reconstruction. All tears were longitudinal and positioned mainly in the posterior horn of the medial meniscus. A total of 70 T-Fixes were used with an average of 3 per patient (range 2-7). Only 4 T-Fixes (6%) were unsuccessfully placed, and this occurred early on in the series in 4 patients. In 90% of the patients, the postoperative activity levels returned to preoperative levels, and the clinical symptoms had either resolved or were experienced at a higher level of activity. The T-Fix device was relatively easy to use and could be reliably placed in the meniscus. Postoperatively, there were no complications directly associated with the device. However, further studies are needed to confirmed these results in a long-term follow-up in a larger patient population. PMID- 9228312 TI - Meniscal injury in children: long-term results after meniscectomy. AB - Twenty-four children who underwent open total meniscectomy were reviewed and followed-up for an average of 16.1 years (range 6-33 years). There were 7 boys and 17 girls aged between 7 and 16 years (average 12.5 years). Excellent and good results were noted in 62.5% of the patients. Eighty-seven of the children showed radiographic degeneration of the knee joint. This study in a Chinese population indicates that meniscectomy is not a benign procedure in children, and total meniscectomy should be avoided as far as possible. PMID- 9228313 TI - Strength of different meniscus suturing techniques. AB - We measured and compared the primary stabilities of five different meniscal suturing techniques. The techniques tested were horizontal mattress, vertical mattress, knot-end, vertical, and vertical loop. Twenty bovine medial menisci were cut to simulate peripheral longitudinal tears and repaired with one of the five suture techniques. Then the two parts of the meniscus were pulled using the Instron Tensometer until failure occurred. Knot-end techniques gave inferior results (mean ultimate failure strength 64 +/- 5 N) compared with the other techniques. Vertical mattress failed at 130 +/- 3 N, vertical loop at 128 +/- 4.5 N, horizontal mattress at 98 +/- 5 N and vertical suturing at 136 +/- 2.7 N. This study shows the superior mechanical characteristic of the vertical suturing technique. PMID- 9228315 TI - Histological and ultrastructural evaluation of Leeds-Keio ligament six years after implant. A case report. AB - We examined by light and electron microscopy study a Leeds-Keio ligament removed from a patient 6 years and 4 months after implant following rupture. The new ligament presented an outer capsule made up of bundles of collagen fibres running mainly perpendicular to the long axis of the ligament. Septa were seen emerging from the capsule and composed of bundles of collagen fibres surrounding the bundles of Dacron fibres. Each thread of Dacron was surrounded by a layer of connective tissue containing periodic acid-Schiff (PAS)-positive cells. The bundles of collagen fibres making up the outer capsule, the septa and the layer of connective tissue surrounding the Dacron threads were positive for anti-type I collagen antibody. The rehabitated Leeds-Keio ligament presented a specific organization at the septa zone, showing a layer of collagen fibrils alternating with a layer of cells. Our remodelling findings suggest a shoelace effect of the artificial ligament. On the other hand, the presence of type I collagen could be responsible for the good functional behaviour of this composite system. In conclusion, the factors that play an important role in determining this remodelling process and its mechanical function are unknown. PMID- 9228314 TI - Pullout strength of tibial graft fixation in anterior cruciate ligament replacement with a patellar tendon graft: interference screw versus staple fixation in human knees. AB - The endoscopic single incision technique for anterior cruciate ligament (ACL) reconstruction with a femoral half-tunnel may lead to a graft/tunnel mismatch and subsequent protrusion of the block from the tibial tunnel. The typical tibial fixation with an interference screw is not possible in these cases. Fixation with staples in a bony groove inferior to the tunnel outlet can be used as an alternative technique. Current literature does not provide biomechanical data of either fixation technique in a human model. This study was performed to evaluate the primary biomechanical parameters of this technique compared with a standard interference screw fixation of the block. Fifty-five fresh-frozen relatively young (mean age 44 years) human cadaver knee joints were used. Grafts were harvested from the patellar tendon midportion with bone blocks of 25 mm length and 9 mm width. A 10-mm tibial tunnel was drilled from the anteromedial cortex to the center of the tibial insertion of the ACL. Three different sizes of interference screws (7 x 30, 9 x 20, 9 x 30 mm) were chosen as a standard control procedure (n = 40). For tibial bone-block fixation the graft was placed through the tunnel, and the screw was then inserted on the cancellous or the cortical surface, respectively. Fifteen knees were treated by staple fixation. A groove was created inferior to the tunnel outlet with a chisel. The bone block was fixed in this groove with two barbed stainless steel staples. Tensile testing in both groups was carried out under an axial load parallel to the tibial tunnel in a Zwick testing machine with a velocity of 1 mm/s. Dislocation of the graft and stiffness were calculated at 175 N load. Maximum load to failure using interference screws varied between 506 and 758 N. Load to failure using staples was 588 N. Dislocation of the graft ranged between 3.8 and 4.7 mm for interference screw fixation and was 4.7 mm for staples. Stiffness calculated at 175 N load was significantly higher in staple fixation. With either fixation technique, the recorded failure loads were sufficient to withstand the graft loads which are to be expected during the rehabilitation period. Staple fixation of the bone block outside of the tunnel resulted in a fixation strength comparable to interference screw fixation. PMID- 9228316 TI - Quantitative gait analysis in patients with medial patellar instability following lateral retinacular release. AB - Medial patellar instability is a known possible complication following lateral retinacular release. Insufficient passive structures, muscle imbalance and an over-release of the lateral retinaculum with resection of the vastus lateralis tendon have been implied as a major cause of this problem. We report about the findings of quantitative gait analysis consisting of video recordings, three dimensional motion analysis, dynamic electromyography and sampling of the ground reaction forces in two patients with medial patellar subluxation. Documentation of the gait functions together with observation of the patellar translation revealed the timing of the occurrence of the instability during level gait: a normal gait pattern with a sufficient quadriceps mechanism and a centred patella was seen during loading (weight acceptance), while the quadriceps muscle was active. Abnormal medial translation of the patella was observed during unloading of the leg while the knee was bending in preparation for the swing phase. This is a phase in the gait cycle when the quadriceps muscle is silent and the patella position is guided by the passive structures only. This finding weakens the argument of muscle imbalance as a cause for the patellar instability and stresses the importance of well balanced passive structures. This explains why a muscular rehabilitation programme is likely to fail as long as the passive structures allow the instability to occur. PMID- 9228317 TI - Long-term results of the treatment of severe osteoarthritis and rheumatoid arthritis with 193 total knee replacements. AB - We undertook a clinical and surgical study with evaluation of the long-term results (average 5 years, range 1-9 years) of 193 stabilized posterior cemented total knee replacements (TKRs) type Insall-Burstein in patients with severe osteoarthritis (OA) and rheumatoid arthritis (RA), carried out consecutively by the same surgeon from January 1986 to January 1995, at our COT Unit at the Central Military Hospital "Gomez Ulla" in Madrid, in collaboration with the Departments of Traumatology and Morphological Sciences of the University of Alcala de Henares (Madrid). The principal purpose was to examine the success rate of this type of prothesis implanted during primary surgery, according to severity of the case. Six methodology protocols were produced in this study (exploratory, surgical technique in primary surgery, revision surgery, rehabilitation, evaluation and clinical revisions), and satisfactory statistical results (SPSS/PC+) were obtained with the three scales of evaluation: Harris Galante, The Hospital for Special Surgery and Knee Society. These were: 90.5% +/- 0.8% excellent and good results for the OA series, and 83.9% +/- 5% for the RA series. The analysis of survival after long-term monitoring was also statistically significant, with a 96.95% survival rate. Complications arose in 3.10% of cases (6 revisions: 4 aseptic loosenings and 2 loosenings due to infection), which were treated with a constrained prosthesis, and in cases of infection with an arthrodesis. After undertaking a comparative study with other series, we conclude that the Insall-Burnstein stabilized posterior total knee prosthesis, is an excellent Primary replacement associated with long-term survival in patients with a severe degree of articular destruction and functional incapacity, and we give some specific recommendations to reduce complications. PMID- 9228318 TI - Bone bruises detected by magnetic resonance imaging following lateral ankle sprains. AB - Although bone bruises have been well described in the knee joint, little is known about their presence in the ankle joint. The present study attempted to document the association of bone bruises with lateral ankle sprains. Magnetic resonance (MR) images were obtained from 60 consecutive patients with lateral ankle sprains between April 1994 and June 1995. There were 29 men and 31 women, aged on average 25 years (range 12-68 years). All of the patients presented within 3 weeks of the sprain. MRI examinations were done within 3 weeks of the injury in 15, after 3-6 weeks in 21, and after 6-8 weeks in 24 cases. There were 28 first-time sprains, while 32 patients had suffered one or more sprains before the most recent one. Plain radiographs showed no evidence of osseous abnormality. Following the conventional MRI examination, magnetic resonance arthrography (MRA) was done by injecting 2 mM of gadolinium diethylene triamine penta-acetic acid (DTPA) into the joint under fluoroscopic control, and the same images were obtained again to search for ligamentous lesions. A total of 11 bone bruises were detected in 10 ankles. In this group of patients, there were 5 men and 5 women aged on average 27 years (range 12-50 years). Four MRI examinations were done within 3 weeks, while six were done 3-6 weeks after the injury. One ankle which had suffered one previous sprain and complete ruptures of anterior talofibular ligament (ATFL) and calcaneofibular ligament (CFL) had two lesions (talus and navicula). In another recurrent case with complete ATFL and CFL ruptures, the lesion was found in the calcaneus. The remaining eight lesions were in the talus in eight ankles. The ligamentous lesions in these ankles included three complete ATFL and CFL ruptures, and four isolated ATFL ruptures; in one ankle there were no ligamentous lesions. The location of talar bruises was medial in six and lateral in three ankles. The incidence of bone bruises associated with isolated ATFL lesions was 16% (4/25). With combined ATFL and CFL lesions the incidence was 50% (5/10). The incidence of ankles with bone bruises and first-time and recurrent sprains was 7% (2/28) and 25% (8/32), respectively. The occurrence of bone bruises should be kept in mind following ankle sprains. Their clinical significance in the long term remains to be determined. PMID- 9228319 TI - Gene transfer to the patellar tendon. AB - Growth factors have the potential to enhance native repair responses in ligamentous lesions. However, methods for applying these cytokines to sites of injury for extended periods are lacking. We suggest that local transfer of genes which encode the relevant healing factors merits investigation as a potential solution to this problem. In the present study, the retroviral vectors MFG lacZ and BAG lacZ neo(r) and adenovirus LacZ were evaluated for their ability to deliver genes to cells of ligamentous origin. The posterior and anterior cruciate ligaments, medial collateral ligament, semitendinosus tendon and patellar tendon were harvested from New Zealand white rabbits. Cells grown from these tissues were then investigated for their susceptibility to genetic alteration by these vectors in vitro. Based upon their ability to convert cells in culture to a lacZ(+) phenotype, adenovirus was the most effective vector in short-term experiments. However, expression was transient. Although retrovirus gave lower initial transduction efficiencies, the percentage of transduced cells could be increased by the use of the selectable marker gene neo(r). In an in vivo marker study, we injected adenovirus into the rabbit patellar tendon. Transduced cells could be observed preferentially in the subsynovial layer at a declining frequency over a 6-week period. The allogeneic transplantation of in vitro retrovirally transduced fibroblasts into the patellar tendon resulted in a greater number of transduced cells. Although the number of lacZ(+) cells declined with time, positive cells were still present 6 weeks after transplantation. Furthermore, the transplanted cells, unlike cells transduced in situ with adenovirus, migrated from the injection site and integrated into the crimp of the tendon. PMID- 9228320 TI - Meniscal allografts: cryopreservation vs deep-frozen technique. An experimental study in goats. AB - Meniscal transplantation was performed in two groups of 15 adult goats each, using cryopreserved (group I) and deep-frozen (group II) allografts. Animals were killed at 2 weeks, 1, 3, 6 and 12 months, and a gross, histological and biochemical (water and glycosaminoglycan) evaluation of the menisci was performed. The allografts of both groups showed a normal gross appearance and had in most cases healed at the horn attachments and at the peripheral capsular tissue with a dense scar tissue and no signs of rejection. Histological analysis showed that at 2 weeks in group I the cell number was decreased compared with the controls, and the cells were mainly distributed in the superficial layers. In group II at 2 weeks, only a few cells were present at the peripheral attachment of the menisci. At 1 month in both groups, the cell repopulation can be seen extending from the peripheral area to the superficial layers. Cell proliferation and vascularization are particularly evident in both groups in the 3-month samples. At 6 months and 1 year the grafts can be seen to be completely remodelled and morphologically similar to normal menisci in both groups. Biochemical analysis showed in both groups an increase in water content and a progressive decrease in the concentration of glycosaminoglycans. At 1 year in both groups, there were moderate degenerative changes in the articular cartilage of the tibial plateau, which were more evident in the area of exposed cartilage than in that covered by the meniscus. These results suggest that there are no significant differences between the cryopreserved and deep-frozen grafts, and that even if cryopreservation makes it possible to maintain a partial cell viability in the tissue, this does not seem to improve the morphological and biochemical characteristics of the graft. PMID- 9228322 TI - Effects of ramipril on the hormone concentrations in serum of hypertensive patients. AB - The effects of the angiotensin-converting enzyme inhibitor ramipril on thirteen endocrinological tests were evaluated. These tests comprised serum follitropin, lutropin, prolactin, thyrotropin, free thyroxine, total thyroxine, free triiodothyronine, parathyrin, cortisol, testosterone, sex hormone binding globulin, androstenedione and dehydroepiandrosterone sulphate. Eleven hypertensive outpatients, 9 men and 2 women, treated at the department of internal medicine in Turku University Central Hospital, received 5 mg of ramipril once a day for the study period of four weeks. The above mentioned endocrinological tests were performed before and at the end of the ramipril treatment. Ramipril decreased the value of free thyroxine statistically significantly, p = 0.011, from the mean value of 17.1 pmol/l to the mean value of 16.0 pmol/l when measured with Amerlex-MAB* free thyroxine kit. The mean within subject difference was -1.10 pmol/l with a 95% confidence interval of -1.87 - 0.33 pmol/l. With the AutoDELFIA free thyroxine kit and with the reference method dialysis+RIA no effect was detected. Other endocrinological tests examined were not affected by ramipril. Since the decreasing effect of ramipril on free thyroxine was detected only with Amerlex-MAB* but neither with AutoDELFIA nor with dialysis+RIA, the effect was concluded to be analytical. The underlying mechanism and the component ultimately interfering with the analysis is unknown. PMID- 9228323 TI - Glutathione and nitric oxide concentrations in glutamine-infused rabbits with intestinal ischaemia/reperfusion. AB - Intestinal ischaemia/reperfusion causes formation of reactive oxygen intermediates which lead to mucosal cell injury. Glutathione, a scavenger of reactive oxygen intermediates, protects tissues from reactive oxygen intermediate mediated cell injury. Nitric oxide is a lipophilic gas and its synthesis is stimulated by ischaemic conditions. In this experimental study, we aimed to investigate the role of i. v. L-glutamine infusion on mucosal tissue glutathione and serum nitric oxide concentrations in intestinal ischaemia/reperfusion. External jugular vein of albino rabbits was cannulated with catheter and infused with normal saline at 4 ml/h. After 3 days, they were randomly divided into two main groups. Group 1 (n = 30) received i. v. normal saline alone, group 2 (n = 30) received normal saline + 205 mmol/l glutamine at 4 ml/h for 24 hours. Next, mucosal glutathione and serum nitric oxide concentrations were measured after 0, 30, 60 min of ischaemia/60 min of reperfusion. Basal glutathione concentrations were similar in normal saline alone and normal saline + 205 mmol/l glutamine infusion groups (p > 0.05). At 30 and 60 min of ischaemia/60 min of reperfusion, glutathione concentrations were significantly lower in normal saline-infused rabbits compared to the normal saline + 205 mmol/l glutamine-infused rabbits (p < 0.05). In addition, serum nitric oxide concentrations were found to be significantly increased in rabbits 30 and 60 min after ischaemia/reperfusion when compared to mean basal nitric oxide concentrations obtained from control animals. However, the normal saline + 205 mmol/l glutamine group had lower serum nitric oxide concentrations than did the normal saline alone group. In conclusion, this study revealed that intestinal mucosal glutathione concentrations were significantly higher in glutamine-receiving rabbits than in non-receiving ones. Additionally, it was shown that nitric oxide concentrations increased in ischaemia both in normal saline alone and normal saline + 205 mmol/l glutamine receiving groups, while this increase in nitric oxide was more prominent in the normal saline alone group (p < 0.01). These findings show that glutamine supplementation may protect the small intestine from ischaemia/reperfusion injury and may play a regulatory role in the biosynthesis of nitric oxide. PMID- 9228324 TI - Effect of serum amyloid A on cellular affinity of low density lipoprotein. AB - Serum amyloid A, an apolipoprotein of high density lipoproteins, is also present to a lesser degree in low density lipoproteins and is co-localized with apolipoprotein B in atherosclerotic lesions. This study examined the effect of serum amyloid A on cellular affinity of low density lipoprotein in vitro. 125I labelled low density lipoprotein, when loaded with recombinant serum amyloid A1 (acute phase isotype) or recombinant serum amyloid A4 (constitutive isotype), had enhanced binding to both human skin fibroblasts and a murine macrophage cell line, J774, while its degradation was slightly increased in both cells. The binding of oxidized low density lipoprotein to J774 cells was also enhanced by addition of recombinant serum amyloid A1 or serum amyloid A4, and degradation of oxidized low density lipoprotein was moderately enhanced by recombinant serum amyloid A1. The effects of recombinant serum amyloid A on cellular binding of labelled low density lipoprotein were not competed by non-labelled low density lipoprotein and were diminished in the presence of high density lipoprotein. These findings suggest that serum amyloid A in low density lipoprotein may promote association of low density lipoprotein with cells by non-specific adsorption, and high density lipoprotein may prevent such interactions by removal of serum amyloid A. PMID- 9228325 TI - Effects of administration of antioxidants in acute intermittent porphyria. AB - In order to elucidate the question of free radical involvement in acute porphyric crisis, antioxidants were administered to two acute intermittent porphyria patients with long-standing recurrent attacks. Clinical condition and urinary excretion of porphyrins and porphyrin precursors were monitored before, during and after an eight week therapy with daily doses of vitamin E, beta-carotene, ascorbic acid, selenium, vitamin Q, acetylcysteine, mannitol and carnitine. Blood cell trace element profiles were followed. The administration of the compound antioxidant formula was found not to further impair the clinical or biochemical conditions of the patients but the incidence of the recurrent crises or the severity of the symptoms were not positively affected. Aberrant blood cell trace element profiles with increased granulocyte manganese were normalized during treatment, on cessation of the therapy again resuming the abnormal pretreatment patterns, which may suggest an origin in oxidative stress. No correlation was observed between the concentration of granulocyte manganese and the excretion of 5-aminolaevulinic acid. Indications for participation of this porphyrin precursor in a radical generating process leading to generalized mitochondrial superoxide dismutase induction, as conceivably signalled by increased intracellular manganese, were thus not obtained. The failure to note a clinical response to antioxidant therapy may be due to factors dependent upon dosage of, or interaction between, the antioxidant compounds given, or on restricted bioavailability of the antioxidants at critical anatomical sites, and does not per se invalidate the model of acute porphyria as a hyperoxidative condition. PMID- 9228327 TI - Transient alkaline hyperphosphatasaemia in an adult: biochemical peculiarities. AB - We report on a 27-year-old healthy female with transient hyperphosphatasaemia of adulthood (it is the eighth case ever recorded). A maximum alkaline phosphatase activity of 1950 U/l, 11-fold the upper reference limit, was measured. The activity normalized within 11 weeks. Electrophoresis revealed the typical pattern for alkaline phosphatase isoenzymes observed in transient hyperphosphatasaemia of infancy: a fast-migrating liver isoenzyme and a bone isoenzyme. Contrary to the findings in transient hyperphosphatasaemia of infancy the liver isoenzyme did not precipitate with wheat-germ lectin whereas the bone isoenzyme partially bound to lectin. Biochemical features of transient hyperphosphatasaemia in an adult may be different from those in infancy. Recognition of an atypical pattern could help avoid unnecessary extensive investigations. PMID- 9228326 TI - Hen egg yolk antibodies purified by antigen affinity under highly alkaline conditions provide new tools for diagnostics. Human intact parathyrin as a model antigen. AB - Hen egg yolks have been recognized as convenient source for specific antibodies, although their utility in diagnostic applications has been hampered by the lack of efficient purification methods. In the present study, anti-human parathyrin antibodies were raised in rabbits, hens and mice using synthetic human parathyrin peptide (1-84) as an antigen. The antibodies were affinity purified with amino- (1-30) and carboxy- (49-89) terminal peptides of parathyrin under highly alkaline elution conditions, and were evaluated for their diagnostic value in different combinations in immunoenzymometric assay (IEMA) format. A pair of hen egg yolk antibodies were subjected to further methodological validation in the IEMA that was constructed with the N-terminal capture and C-terminal detection antibodies. The synthetic intact human parathyrin (1-84) peptide served as a standard. This within-day IEMA procedure turned out to be sensitive and it correlated well with the two independent intact parathyrin immunoradiometric assays. As shown in the present study, the immuno affinity purification with the highly alkaline elution conditions provides an efficient method for utilization of hen egg yolk antibodies. This is the first report on an application making use of a combination of two hen egg yolk antibody preparations in measuring a homogeneous protein in human serum. PMID- 9228328 TI - Assay of serum/plasma beta-N-acetylhexosaminidase isoenzymes by heat inactivation using a continuous spectrophotometric method adapted to a centrifugal analyzer. AB - Activity of serum/plasma beta-N-acetylhexosaminidase (EC 3.2.1.52) was determined by means of a continuous spectrophotometric method using 3,3' dichlorophenylsulphonphthaleinyl-N-acetyl-beta-D-glucosaminid e as substrate, with very satisfactory results. Incubation of an undiluted aliquot (1 ml) of samples at 52 degrees C for 8 hours with an adjusted pH 5.5-6.0 provoked only the inactivation of isoenzyme A, thus allowing the evaluation of beta-N acetylhexosaminidase isoenzyme composition. In 25 serum samples from control subjects and pregnant women, a good correlation between the percentage of isoenzyme B obtained by this procedure and the fluorimetric assay of O'Brien et al. (New Engl J Med 1970; 273:15-20) was found (r = 0.983, S(yx) = 1.51), with no statistically significant difference between the means (43.2 vs 42.8%). In 84 healthy adult subjects, an average value of 30.3% for the proportion of isoenzyme B was obtained, with an interval of 25.4-35.0%, in agreement with results reported by other authors. PMID- 9228329 TI - Within-subject variation of carcinoembryonic antigen in colorectal cancer -- application of reference of change values and individual reference ranges to patient follow-up. AB - Dynamic interpretation of results is an alternative approach to the conventional cut-off procedure. Reference change limit is a valuable reference point to interpret dynamically tumour marker values when only very few serial results can be obtained from a patient after treatment. In this paper, a reference change limit of 2.0 microg/l was estimated for carcinoembryonic antigen in patients with complete remission of colorectal cancer. This figure means that a difference greater than 2.0 microg/l has at least a chance of being statistically significant (at 0.05 probability). As complementary information to the reference change limits, with more than four successive results, a simple time series model can be used to obtain predictive limits for the next observation. PMID- 9228330 TI - Cumulative troponin T release after acute myocardial infarction. Influence of reperfusion. AB - For troponin T a characteristic biphasic change in the plasma time-concentration curve has been described, especially in patients with early reperfusion after thrombolytic therapy. As troponin T is bound to myofibrillar structures, treatment strategy or treatment outcome could influence the cumulative plasma release of this protein in a different way compared to the cumulative release of free cytoplasmic cardiac enzymes. The present study is the first study comparing the total quantity of troponin T released by the heart during the first 168 hours after acute myocardial infarction, both in patients treated with thrombolytic therapy (n = 16) and in patients not treated with thrombolytic therapy (n = 7). On the basis of clinical symptoms and coronary arteriogram within 24 hours, the patients treated with thrombolytic therapy were divided into two groups, reperfused (n = 9) and non-reperfused (n = 7). In the patients not treated with thrombolytic therapy, absence of spontaneous early reperfusion was judged only from clinical symptoms. Cumulative troponin T release into plasma was compared to the cumulative release of the cytoplasmic cardiac enzymes creatine kinase (EC 2.7.3.2) and hydroxybutyrate dehydrogenase (EC 1.1.1.27). Cumulative release, i. e., infarct size, was calculated using a two-compartment model for circulating proteins. Mean tissue contents, per gram wet weight, of 156 U/g for hydroxybutyrate dehydrogenase, 2.163 U/g for creatine kinase and 234 microg/g for troponin T, were used to express infarct size in gram-equivalents of healthy myocardium per litre plasma (g-eq/l). Release rates were represented by the ratio of cumulative quantities released in 10 hours and 72 hours for creatine kinase and hydroxybutyrate dehydrogenase and in 10 hours and 168 hours for troponin T. CONCLUSIONS: - Plasma time-concentration curves and release rates of troponin T in patients treated with thrombolytic therapy showing reperfusion differ significantly from those of patients not treated with thrombolytic therapy, showing no reperfusion. - Creatine kinase and hydroxybutyrate dehydrogenase release is completed within 72-100 hours in all patients, whereas troponin T release still continues after 168 hours. - Cumulative troponin T release at 168 hours is only a fraction (around 8%) of cumulative cytoplasmic enzyme release and the percentage released is not influenced by the treatment strategy or outcome, i. e., vessel patency. - Although troponin T release is only a fraction of the cumulative enzyme release (infarct size) there is a highly significant correlation between both, independent of the treatment strategy or treatment outcome. PMID- 9228331 TI - Epitestosterone in human blood and prostatic tissue. AB - Epitestosterone, a C19-steroid with anti-androgenic activity, was determined in the plasma of 234 boys and men from the ages of 6-86 years, and in the prostate tissue of 15 men 55-82 years of age. It was documented that, while in adulthood the concentration of epitestosterone is about ten times lower than the concentration of testosterone, in the pre-pubertal period the level of epitestosterone is similar or even higher than that of testosterone. In the hyperplastic prostate tissue the content of epitestosterone is comparable to that of androstenedione, it is about twice as high as the content of testosterone and approximately half that of the content of dihydrotestosterone. At least in the case of pre-pubertal boys and in the prostatic tissue it is therefore possible to include epitestosterone into consideration as a regulatory factor for the androgen-dependent events. PMID- 9228332 TI - The additional value of free prostate specific antigen to the battery of age dependent prostate-specific antigen, prostate-specific antigen density and velocity. AB - This study describes the value of using the fraction of free prostate-specific antigen as a further marker in the early detection of prostate cancer. This newly introduced marker is compared to the usual battery of age-dependent total prostate-specific antigen, prostate-specific antigen density (microg/l x g tissue) and prostate-specific antigen velocity (microg/l x year). Determination of total prostate-specific antigen and free prostate-specific antigen was performed on fresh serum samples obtained from 3470 symptomatic patients aged 45 80 attending the Urology Clinics, or their General Practitioners. Among them, 310 patients had total prostate-specific antigen above the age-dependent cut-off, and/or free/total prostate-specific antigen under 11%, with different prostate specific antigen densities and velocities. Only 147 patients complied to undergo biopsy: in 72 of those patients, benign prostatic disease was histologically confirmed, while in 75 patients primary prostate cancer was histologically confirmed. Total and free prostate-specific antigen levels were determined using the third generation DPCs prostate-specific antigen assay performed on the Immulite automated immunoassay instrument. Total prostate-specific antigen age reference values were adopted from Oesterling et al. (J Am Med Ass 1993; 270:860 4); the prostate-specific antigen density was considered suspicious of prostate cancer if it was greater than 0.15 microg/l prostate-specific antigen per gram tissue (Seaman et al. Urol Clin N Am 1993; 20:653); prostate-specific antigen velocity greater than 0.75 microg/l x year (Carter et al., J Am Med Ass 1992; 267:215) was considered suspicious for prostate cancer. Of the 147 patients, 75 had prostate cancer and 72 had benign prostatic hypertrophy. The difference between prostate cancer and benign prostatic hypertrophy was significantly reflected only by free/total prostate-specific antigen and prostate-specific antigen velocity. These parameters also provided the best sensitivity and specificity. Only these parameters proved to be significant when using a backwards logistic regression model (prostate-specific antigen velocity, p = 0.007 odds ratio 2.782; free/total prostate-specific antigen %, p = 0.016 odds ratio 2.678). Combinations of various parameters became significant when including free/total prostate-specific antigen, increasing prostate cancer detection to 88%. We conclude that free/total prostate-specific antigen is the most significant among prostate-specific antigen quantities (total age-dependent prostate-specific antigen, prostate-specific antigen density and prostate specific antigen velocity). Adding this parameter to other prostate-specific antigen parameters improves the discrimination between prostate cancer and benign prostatic hypertrophy for the population at risk. PMID- 9228333 TI - Reliability of serum separator blood drawing tubes under routine conditions. PMID- 9228334 TI - An economic argument in favour of endoscopic third ventriculostomy as a treatment for obstructive hydrocephalus. AB - This project was undertaken to examine the health resource implications of performing endoscopic third ventriculostomy as an alternative to CSF shunting in appropriate patients. We carried out a retrospective study of case records and X rays of patients shunted de novo at the INS, Glasgow for the two year period 1990 1991. We identified all those patients who would have been suitable for endoscopic third ventriculostomy and examined the shunt complications and extra days in hospital required by these patients. A total of 150 new shunts was inserted during the two year period. Of these, 23 patients (15%)were judged suitable for endoscopic third ventriculostomy as an alternative to CSF shunting. Eight out of 23 patients required a total of 29 repeat operations and an extra 230 days in hospital due to shunt complications. Assuming an 80% success (shunt free) rate for endoscopic third ventriculostomy, we calculate that 9 operations and 74 bed days per year could be saved by using this technique. We conclude that in units undertaking a large number of CSF shunt insertions, investment in neuroendoscopic equipment, training, and expertise has the potential to release significant resources for other uses. PMID- 9228335 TI - Annual risk for the first hemorrhage from untreated cerebral arteriovenous malformations. AB - To estimate the annual risk for the first hemorrhage, survival and life table statistics were used to analyze data from 2262 patients with cerebral arteriovenous malformations (AVM). We found that the risk for hemorrhage increases with increasing age. A validity test revealed, however, that life table statistics used on the total patient material underestimated the annual risk for hemorrhage, especially for patients 20-50 years of age. A method based on the fact that the distribution of the time at risk until the initial hemorrhage (= age at first rupture) reflects the risk for hemorrhage in untreated AVM was therefore also employed. The analysis yielded three conclusions: 1) the annual risk of hemorrhage increases with age; 2) small AVM are less prone to rupture; and 3) the risk of hemorrhage is higher in women during their fertile years as compared to males in the same age group. The risk related to age, AVM size and location assessed by survival statistics in the subgroup of patients with a known date for the initial hemorrhage gave similar results. PMID- 9228336 TI - Enhancing neurosurgical endoscopy with the use of 'virtual reality' headgear. AB - Widespread use of neurosurgical endoscopy has been hampered by the necessity of looking up from the surgical field to view the endoscopic image on a video monitor. Here is demonstrated a simple, lightweight headpiece with a small, built in video monitor that reflects the video image to the dominant eye, thus allowing the operator to continuously observe the surgical field either via peripheral vision or via the non-dominant eye. Successful use of the device is documented here in a minimally invasive fenestration of an arachnoid cyst into the lateral ventricle via a flexible endoscopic procedure. PMID- 9228337 TI - Three-dimensional computer-assisted stereotactic-guided microneurosurgery combined with cortical mapping of the motor area by direct electrostimulation. AB - TIM (Zeppelin Chirurgische Instrumente GmbH, 82 049 Pullach, Germany) is a tomographic imaging system which enables surgeons to visualize the pathologic lesions three dimensionally in relationship to the surrounding structures. The distance and the angle between the pathologic lesion and the anatomical and/or bony landmarks as well as the volume of the mass lesion can be measured. Therefore an accurate localization of the lesion is possible with this technique. It is very applicable for planning of surgery on skull base tumors. The surgical procedure for small and well-defined, intrinsic pathologic deep-seated brain lesions, however, becomes much easier by using the stereotactic techniques of this system. The target point and the direction brain-surface-to-lesion can be determined within seconds. Before the aiming probe is inserted to the target, the cortical motor area is mapped by direct electrical stimulation. The approach can be varied depending on the results of these neurophysiologic investigations of the brain surface. The dissection is made along the aiming probe up to the target point. Because of the fixation of the brain with the needle, a brain shifting due to the dissection as well as to CSF release is diminished. Forty patients with deep-seated intracerebral lesions were operated on during a 13 months period by these combined techniques in our service. Using this technique, we never made a negative exploration. In all but three patients, total removal of the mass lesion was achieved. Permanent neurological deficits were observed in two patients only. In our opinion, this combined imaging and neurophysiological technique is easy to perform, and of major benefit for the patients due to its accuracy and is preferable in comparison with other single computer localizer techniques without neurophysiological monitoring. PMID- 9228338 TI - Intracavitary irradiation with colloidal phosphorus-32 for treatment of an arachnoid cyst: a new approach. AB - Intracavitary irradiation is reported as an additional treatment for intracranial arachnoid cysts that do not communicate with the subarachnoid space. A 46-year old woman with a large suprasellar arachnoid cyst that had enlarged over a 4-year interval presented with new onset headaches. Stereotactic intracavitary irradiation was performed using colloidal phosphorus-32 as an alternative to craniotomy or insertion of a shunt. Total regression of the cyst occurred within one month with resolution of her headaches. After three years of follow-up, imaging studies showed only an empty sella appearance, and no cyst recurrence. No early or delayed morbidity occurred. Intracavitary irradiation may be a useful therapy for arachnoid cysts, in an attempt to alter the biology of the cyst lining. PMID- 9228339 TI - Contralateral massive acute subdural collection after endoscopic third ventriculostomy - a case report. AB - An uncommon case of contralateral massive subdural collection developing acutely after a neuroendoscopic third ventriculostomy is presented. The patient developed a cardiorespiratory arrest in the recovery room where he could be successfully resuscitated. He made a complete recovery following aspiration of the subdural collection. PMID- 9228340 TI - Glabellar approach: simplified midline anterior skull base approach. AB - As a simplified microsurgical technique for lesions at the midline anterior skull base, a glabellar approach through a small incision (5 cm) between the eyebrows crossing the nasion was developed in four cadaver dissections. To determine the ideal positioning of the patient, the angle of the surgical trajectory was measured in the sagittal plane. In an effort to make an operation simple and accurate through this limited exposure, measurements were made in distance from the midline nasion to various intradural structures. Average distance from the midline point of the nasion (MPNa) to the midline of the tuberculum sella was 6.37 +/- 0.29 cm, 6.98 +/- 0.26 cm to the midline of the optic chiasm, and 8.00 +/- 0.11 to the lamina terminalis. In addition, measurements to other various anatomical landmarks were made. The angle of the line drawn from the MPNa to the midline of the tuberculum sella was 5.2 +/- 1 degrees against a line drawn between the lateral canthus and the tragus (LC-T) in the sagittal plane. Based upon this study, the positioning of the patient's head would be better if the LC T line is positioned at 25 degree extension when the operating microscope is set at 20 degree inclination. Within 6 to 8 cm in depth from the MPNa, important landmarks are exposed without brain retraction. If surgical instruments are marked with ruler calibration, the depth of the surgical instruments will suggest the anatomical location. This glabellar approach has been used in three patients successfully. A brain retractor was not necessary and not used during the operations. This technique provides key exposures to the midline anterior skull base. PMID- 9228341 TI - The anatomical variations of sylvian veins and cisterns. AB - The anatomical variations of sylvian vein and cistern were investigated during the pterional approach in 750 operative cases with different pathologies. All patients were operated on at the Neurosurgical Department of Ataturk University Medical School, Erzurum, Turkiye. The patients underwent surgery for the lesions necessitating the right or left pterional approach. The findings were recorded during surgical intervention and observed through the operative sketches of the pathologies, the slides, and videotapes of the operations. In our study, we surgically classified the variations of sylvian vein, according to its branching and draining patterns. Type I: The fronto-orbital (frontosylvian), fronto parietal (parietosylvian) and anterior temporal (temporosylvian) veins drain into one sylvian vein. Type II: Two superficial sylvian veins with separated basal vein draining into the sphenoparietal and Rosenthal's basal vein. Type III: Two superficial sylvian veins draining into the sphenoparietal and the superior petrosal veins. Type IV: Hypoplastic superficial sylvian vein and the deep one. Four types of sylvian vein variations were defined as follows. The type I was seen in 52.8% (n = 396), the type II was found in 19.2% (n = 144), type III was recorded in 18.2% (n = 137), and type IV, or hypoplastic and deep form was discovered in 9.8% (n = 73) of patients. The coursing of sylvian vein was in the temporal side (Temporal Coursing) in 62.4 percent of the cases (n = 469), in the frontal side (Frontal Coursing) in 25 % of the patients (n = 187) and in 9 percent of the cases (n = 67) in the deep localization (Deep Coursing). Only 3.6% of the cases (n = 27) showed Mixed Coursing. The variations of the sylvian cisterns were classified into three types, according to the relationships between the lateral fronto-orbital gyrus and the superior temporal gyrus. In Sylvian type, the frontal and temporal lobes are loosely (Sylvian Type A, wide and large) or tightly (Sylvian Type B, close and narrow) approximated on the surface thereby covering the substance of the sylvian cistern. In Frontal Type, the proximal part of the lateral fronto-orbital gyrus herniated into the temporal lobe. In Temporal Type, the proximal part of the superior temporal gyrus herniated into the lateral fronto-orbital gyrus. The variations of the sylvian cisterns in 750 patients with different pathologies, were as follows: in 47.7% (n = 358) Sylvian type A, in 27.2% percent (n = 204) Sylvian type B, in 16.3% (n = 122) frontal type and in 8.8% (n = 66) temporal type. We concluded that venous perfusion discorder of the brain is the most important factor during the pterional approach. Careful intraoperative assessment and protection of the sylvian vein, which is a surgical pitfall, is an indispensable part of the operation. The recognition of the anatomical variations of the sylvian vein and cistern, and the detailed knowledge of the microvascular relationships and the importance of preservation of this vein at that level, will allow the neurosurgeon, believing in the minimally invasive neurosurgical techniques, to construct a better and safer microdissection plan, to save time, and can prevent postoperative neurological deficits. PMID- 9228342 TI - Unilateral hemilaminectomy for the removal of the spinal space-occupying lesions. AB - In this study we have evaluated 40 patients with spinal lesions with respect to the value of unilateral hemilaminectomy. Our case study group included 29 intradural extramedullary, 6 intramedullary, and 5 extradural tumors. The thoracic spinal cord was involved in 17, the lumbar region in 13, and the cervical spinal cord in 10 cases. The mean age of the 20 males and 20 females was 35 (range 6-71). The rationale for choosing a unilateral approach is to preserve musculoligamentous attachments and bony posterior elements as much as possible. We did not observe any complication relating to unilateral hemilaminectomy. The patients were mobilized the following day after surgery or given rehabilitation therapy beginning on the second postoperative day without the use of any external support. At follow-up evaluation, a mean of 32 months postoperatively, none of the patients showed spinal deformity or spinal instability. PMID- 9228344 TI - Methotrexate revisited. PMID- 9228345 TI - Asthma therapy: what's new and is it necessarily better? PMID- 9228348 TI - Is hospice referral ever appropriate in COPD? PMID- 9228347 TI - T.G. Heaton, tuberculosis, and artificial pneumothorax: once again, back to the future? PMID- 9228346 TI - In pursuit of tuberculosis control: civil liberty vs public health. PMID- 9228349 TI - Hypertension and heart failure: the link continues. PMID- 9228350 TI - The effect of regular salbutamol on lung function and bronchial hyperresponsiveness in normal subjects and nonasthmatic atopic subjects with allergic rhinitis. AB - BACKGROUND: The effects of regular inhaled beta-agonist treatment on lung function remain a controversial issue. In an earlier study, the administration of regular inhaled salbutamol resulted in negative changes in FEV1 not only in asthmatics, but also in nonasthmatic atopic subjects. OBJECTIVE: The aim of this study was to confirm these findings and also to examine whether regular inhaled salbutamol would increase bronchial hyperresponsiveness in atopic or normal individuals. DESIGN: The study was a randomized, double-blind, placebo controlled, crossover investigation. PARTICIPANTS: There were two groups: 10 normal healthy subjects (group A) and nine nonasthmatic atopic subjects (group B). INTERVENTIONS: Subjects received inhaled salbutamol or identical placebo for periods of 6 weeks, the dose being increased in stepwise fashion at 2-week intervals. MEASUREMENTS: Changes in lung function were assessed by measuring spirometric values, airways conductance, and changes in complete and partial expiratory flow volume curves. Changes in these parameters following a methacholine challenge were used to assess bronchial hyperresponsiveness. RESULTS: No statistically significant differences in lung function or bronchial hyperresponsiveness were noted to occur as the result of treatment in either group. CONCLUSION: Our results do not support the view that regular inhaled salbutamol in normal or atopic subjects without asthma causes adverse changes in the airways. PMID- 9228351 TI - Effect of age on bronchodilator response in acute severe asthma treatment. AB - STUDY OBJECTIVES: This study was designed to evaluate the effects of age on bronchodilator response to salbutamol in patients with acute severe asthma in the emergency department. SUBJECTS AND METHODS: Sixty-four sequential patients (mean age, 34.2+/-10.7 years) with acute asthma were enrolled in the trial. Using age as a major criterion, we divided the sample in two groups: the young one (age < or = 35 years, n=30) and the older (> 35 years, n=34). All patients were treated with salbutamol delivered with metered-dose inhaler into a spacer device, in a dose of four puffs every 10 min (100 microg per actuation) during 3 h. RESULTS: Mean FEV1 improved significantly over baseline values for both groups (p=0.001). At final disposition, the mean percent of predicted FEV1 was 55.1+/-16.3% in the young group and 58.0+/-20.9% in the older group. There were no significant differences between both groups for FEV1 percent response at any point studied. A significant increase in heart rate over baseline was seen in the older group (p=0.001). Older patients also presented a higher incidence in nausea and tremor. Young and older patients with acute asthma achieved equivalent bronchodilation response to salbutamol. CONCLUSIONS: We concluded that age is not a predictor of response to beta-agonists. PMID- 9228352 TI - Treatment of acute severe asthma with inhaled albuterol delivered via jet nebulizer, metered dose inhaler with spacer, or dry powder. AB - Despite the increasing use of dry powder formulations in the ambulatory setting, there is a paucity of information on the efficacy of this therapeutic modality to treat acute severe asthma. In addition, studies that compared wet nebulization vs metered dose inhalers formulated with chlorofluorocarbon (CFCMDI) attached to holding chambers have yielded discrepant results. Thus, it is unclear which of the three delivery systems would elicit a superior bronchodilator response, particularly in patients with life-threatening asthma. In a prospective, randomized open design, we studied the response to inhaled albuterol (salbutamol) in 27 adult asthmatics presenting to the emergency department (ED) with an FEV1 <30% predicted. Subjects were treated with one of the following regimens (nine subjects in each group): group A, mean (SD) baseline FEV1 of 0.7 (0.2) L, received albuterol solution, 5 mg, via a nebulizer (Puritan-Bennett Raindrop; Lawrenceville, Ga) impelled with oxygen (O2) at 8 L/min; group B, baseline FEV1 of 0.6 (0.15) L, received albuterol, 400 microg, via a CFCMDI attached to a 145 mL valved aerosol holding chamber (Aerochamber; Trudell Medical; London, ON); and group C, baseline FEV1 of 0.6 (0.17) L, received albuterol powder, 400 microg, by another means (Rotahaler; Glaxo; Research Triangle Park, NC). All groups received the respective treatments on arrival in the ED, every 30 min during the first 2 h, and then hourly until the sixth hour. Clinical parameters and FEV1 were recorded on ED admission and 15 min after each dose of albuterol. At the time of ED admission, all patients also received continuous O2 and one dose of I.V. steroids (dexamethasone, 8 mg). The total dose of inhaled albuterol administered during the 6-h treatment was 45 mg of nebulized solution in group A and 3,600 microg of albuterol aerosol and dry powder in groups B and C, respectively. No significant differences were found in the population demographics, baseline FEV1, and arterial blood gas values on air. FEV1 improved significantly in all patients after the 6 h of treatment. The 6-h area under the curve FEV1 improved similarly with the three delivery methods despite differences in the total dose administered. No patient was discontinued during the trial or admitted to hospital and no evidence of cardiovascular adverse events was apparent in any of the study groups. These data support the view that the three delivery methods appear adequate to treat subjects with acute severe asthma. PMID- 9228353 TI - Low-dose methotrexate spares steroid usage in steroid-dependent asthmatic patients: a meta-analysis. AB - STUDY OBJECTIVES: To determine if treatment with low-dose methotrexate spares oral steroids in adult, steroid-dependent, asthmatic patients. DESIGN: Articles identified by computer search were excluded if they (1) did not contain original data relating to the primary question, (2) had no controls, or (3) described subjects younger than 18 years of age. MEASUREMENTS: From each article, the following was abstracted: reference citation; type of control; whether study incorporated a run-in period in which the baseline level of prednisone was reduced to the lowest possible dose; dosage and length of methotrexate therapy; baseline dosage; and type of steroid. Also, the following was determined during both placebo and methotrexate arms of each study: steroid dosage, FEV1, serious side effects, and alteration in level of serum aspartate aminotransferase. RESULTS: Eleven eligible studies were identified. Methotrexate treatment resulted in a decrease in prednisone or prednisolone usage by an average of 4.37 mg/d or 23.7% of the initial dosage. The summary effect size in standard deviations (SDs) was -0.53 (95% confidence interval=-0.29 to -0.77). Subgroup analysis showed that patients treated with prolonged (6-month) therapy with methotrexate, those with low long-term usage of steroids (< or = 20 mg/d), and those whose study design incorporated a run-in period, tended to have the greatest steroid-sparing effects with methotrexate. CONCLUSIONS: Low-dose methotrexate has a significant steroid sparing effect in steroid-dependent asthmatic patients. The greatest effect was evident in patients in whom an effort was made to reduce baseline steroid dosage and in whom methotrexate was used for 24 weeks. PMID- 9228354 TI - A comparison of double-strength beclomethasone dipropionate (84 microg) MDI with beclomethasone dipropionate (42 microg) MDI in the treatment of asthma. AB - STUDY OBJECTIVE: To compare the efficacy and safety of a double-strength formulation of beclomethasone dipropionate (BDP 84) metered-dose inhaler (MDI) with that of beclomethasone dipropionate (BDP 42) MDI in the treatment of chronic asthma. DESIGN: A 28-day, randomized, double-blind, double-dummy, placebo controlled, multicenter study. SETTING: Outpatient. PATIENTS: A total of 423 patients aged 12 to 65 years (mean range, 34 to 36 years) with moderate asthma (FEV1, 50 to 80% of predicted) who required long-term inhaled corticosteroids were enrolled. INTERVENTIONS: Patients were randomized to receive BDP 84, two oral inhalations bid (336 microg/d), BDP 42, four oral inhalations bid (336 microg/d), or placebo. A fourth treatment arm administering BDP 84, eight oral inhalations bid (HD BDP 84; 1,344 microg/d) was also included to determine whether a dose-response relationship could be demonstrated. MEASUREMENTS: Spirometry, clinical observations. RESULTS: The three active treatments were significantly more effective (p < or = 0.01) than placebo at all time points in improving FEV1, the primary efficacy parameter; BDP 42 and BDP 84 were comparable to each other at every time point. Secondary pulmonary function tests (FVC, forced expiratory flow at 25 to 75% of FVC, and peak expiratory flow rate) showed similar results. All three active treatments were well tolerated. A dose response between 336 microg/d and 1,344 microg/d was demonstrated. CONCLUSION: In this well-controlled 28-day study, BDP 42 and BDP 84 were shown to be comparable in efficacy and safety on a microgram-for-microgram basis. PMID- 9228355 TI - Renal dose dopamine does not alter the response to beta-adrenergic stimulation by isoproterenol in healthy human volunteers. AB - OBJECTIVES: To determine if renal dose dopamine (3 microg/kg/min) alters the heart rate (HR) by itself, or if a dopamine infusion alters the HR response to bolus doses of the beta-adrenergic agonist isoproterenol in healthy human subjects. DESIGN: Prospective study. SETTING: Clinical laboratory of a university affiliated academic medical center. SUBJECTS: A total of 15 healthy nonpregnant women and men aged 21 to 44 years. INTERVENTIONS: Subjects were monitored continuously with bedside ECG, pulse oximetry, and ambulatory ECG recording to measure the maximal HR response to separate injections of 10, 20, and 30 ng/kg of isoproterenol, given before, during, and after the infusion of 3 microg/kg/min of dopamine. MEASUREMENTS AND MAIN RESULTS: Dopamine in the absence of isoproterenol did not alter baseline HR significantly (62.7+/-2.2 beats/min without dopamine; 65.4+/-2.2 with dopamine; p=0.15). All three doses of isoproterenol increased HR significantly above baseline, both in the presence and absence of dopamine (p<0.001). Dopamine infusion resulted in a higher HR following isoproterenol only for the 20-ng/kg dose. The incremental increases in HR, defined as the difference between peak HR following isoproterenol and baseline HR, were not increased during dopamine infusion for any of the doses of isoproterenol. Nausea was reported by 5 of the 15 subjects during the dopamine infusion. CONCLUSIONS: In healthy human subjects, infusion of 3 microg/kg/min of dopamine does not significantly increase the HR when combined with beta-adrenergic stimulation using isoproterenol, suggesting neither an additive nor antagonistic interaction between the two drugs. While our study did not demonstrate an increase in HR in healthy subjects, the risk of increasing the chronotropic response to beta adrenergic inotropic medications with "renal dose" dopamine in critically ill patients needs to be investigated. The frequency of nausea during dopamine infusion also may influence consideration of using dopamine to augment splanchnic blood flow and renal function in conscious patients. PMID- 9228356 TI - Bronchial subepithelial fibrosis correlates with airway responsiveness to methacholine. AB - OBJECTIVES: To evaluate the relationships between airway subepithelial collagen deposition and epithelial desquamation with airflow obstruction and hyperresponsiveness in different types of asthma and other respiratory conditions such as chronic cough and allergic rhinitis. DESIGN AND PARTICIPANTS: We compared the histopathologic features observed on bronchial biopsy specimens obtained from 80 subjects: 38 with different types of asthma, 19 with chronic cough, 13 with allergic rhinitis, and 10 normal control subjects. Each subject had a questionnaire on respiratory symptoms and medication needs, measurements of expiratory flows and methacholine responsiveness, allergy skin prick tests, and a bronchoscopy with bronchial biopsies. None of the subjects studied used bronchial anti-inflammatory agents. RESULTS: Different degrees of bronchial subepithelial fibrosis were present in asthmatic subjects, the most intense being observed in occupational asthma; a subepithelial deposition of collagen was also found in subjects with allergic rhinitis, although it was less intense than in asthma and irregularly distributed under the basement membrane. On global analysis, we found a significant correlation between individual provocative concentration of methacholine inducing a 20% fall in FEV1 (PC20) and subepithelial fibrosis intensity (rs=-0.70, p<0.001). The degree of epithelial desquamation was correlated with that of subepithelial fibrosis (rs=0.36, p=0.02) in subjects with normal airway responsiveness, but it was not correlated with the PC20 (rs=0.10, p>0.05). Neither the degree of subepithelial fibrosis nor epithelial desquamation was correlated with the FEV1. CONCLUSION: These results suggest that structural airway changes such as subepithelial collagen deposition may be significant determinants or markers of a process that results in airway hyperresponsiveness. PMID- 9228357 TI - Methacholine challenge testing: safety of low starting FEV1. Asthma Clinical Research Network (ACRN). AB - STUDY OBJECTIVE: The lower limit for the baseline value to initiate methacholine bronchial hyperresponsiveness testing has not been well established. Recommendations have varied from > 1 L to above 80% of predicted. The objective was to determine if an FEV1 < 60% predicted was acceptable. DESIGN: Retrospective analysis of challenges in 88 patients with a baseline FEV1 of < 60% predicted (mean=45.8%; range, 22 to 59%. SETTING: Academic institutions. RESULTS: There were only four individuals whose FEV1 did not return to > 90% of baseline following one poststudy beta2-agonist treatment. All four responded to a second treatment. There were no adverse sequelae following challenge in any individual. Neither age (up to 79 years) nor gender influenced outcome. CONCLUSIONS: In chronic moderate to severe asthma, it appears that bronchial hyperresponsiveness testing can be safely performed even in those patients with a low baseline FEV1. PMID- 9228360 TI - Use of transesophageal echocardiography for diagnosis of traumatic aortic injury. AB - This prospective study was conducted to describe the signs on transesophageal echocardiography (TEE) associated with traumatic aortic injury (TAI). Twenty eight patients with TAI underwent TEE, and they were compared with a control group of 30 thoracic trauma patients without aortic injury. The TEE signs were classified as direct or indirect signs, and the quality of imaging was assessed. Patients' TEE images were compared with their anatomic lesions. The direct signs were thick stripes (n=19), false aneurysm (n=7), aortic dissection (n=6), free edge intimal flap (n=15), aortic wall hematoma (n=2), fusiform aneurysm (n=13), and complete aortic obstruction (n=2). The indirect signs included minor increases in aortic diameter (n=7), impairment of the aortic Doppler color flow (n= 18), and an increase of aorta-probe distance, indicating hemomediastinum (n=23). TEE allowed diagnosis of recently described limited intimal lesions frequently missed by other conventional methods, and permitted rapid diagnosis of complete rupture in which fast degeneration means that more time-consuming methods are not practicable. Significant blurring of the aortic outline was noted in 20% of cases and intraluminal artifacts were observed in 36% of cases, but neither sign impaired accurate diagnosis of TAI. The echocardiographic signs of aortic injury are complex and may be confined to a short section of the aorta. Therefore, examination by a physician highly trained in echocardiography is necessary in such cases. PMID- 9228358 TI - Short-term incarceration for the management of noncompliance with tuberculosis treatment. AB - STUDY OBJECTIVES: To review the use of incarceration for noncompliance with tuberculosis treatment. DESIGN: Retrospective review. SETTING: An urban tuberculosis control program. PATIENTS: Patients treated for active tuberculosis. MEASUREMENTS AND RESULTS: We reviewed the legal basis and practical application of quarantine for active tuberculosis, including the use of incarceration for noncompliance. The records of patients treated at the Denver Metro Tuberculosis Clinic during 1984 to 1994 were reviewed to identify patients who were incarcerated and to evaluate the effectiveness of this intervention. Of 424 cases of tuberculosis, 20 patients (4.7%) were incarcerated for noncompliance; an additional 21 patients (5.0%) were lost to follow-up prior to completing therapy. Incarcerated patients were predominantly men who were born in the United States and had a history of homelessness and alcohol abuse. The median duration of the initial incarceration was 20 days (range, 7 to 51 days). Of the 17 patients released prior to completing therapy, 13 (76%) were compliant with outpatient, directly observed therapy after one or two short-term incarcerations (<60 days); only three patients were incarcerated for the duration of treatment. Overall, 18 of 20 incarcerated patients (90%) were successfully treated. CONCLUSIONS: Approximately 5% of the patients treated through our program were incarcerated for noncompliance; an additional 5% were unavailable for follow-up and would have been candidates for incarceration if found. Homelessness and alcoholism were closely associated with the use of incarceration. Short-term incarceration followed by outpatient, directly observed therapy was relatively successful in the management of this difficult patient population. PMID- 9228359 TI - A cost-effectiveness analysis of directly observed therapy vs self-administered therapy for treatment of tuberculosis. AB - STUDY OBJECTIVES: To compare the costs and effectiveness of directly observed therapy (DOT) vs self-administered therapy (SAT) for the treatment of active tuberculosis. DESIGN: Decision analysis. SETTING: We used published rates for failure of therapy, relapse, and acquired multidrug resistance during the initial treatment of drug-susceptible tuberculosis cases using DOT or SAT. We estimated costs of tuberculosis treatment at an urban tuberculosis control program, a municipal hospital, and a hospital specializing in treating drug-resistant tuberculosis. OUTCOME MEASURES: The average cost per patient to cure drug susceptible tuberculosis, including the cost of treating failures of initial treatment. RESULTS: The direct costs of initial therapy with DOT and SAT were similar ($1,206 vs $1,221 per patient, respectively), although DOT was more expensive when patient time costs were included. When the costs of relapse and failure were included in the model, DOT was less expensive than SAT, whether considering outpatient costs only ($1,405 vs $2,314 per patient treated), outpatient plus inpatient costs ($2,785 vs $10,529 per patient treated), or outpatient, inpatient, and patients' time costs ($3,999 vs $12,167 per patient treated). Threshold analysis demonstrated that DOT was less expensive than SAT through a wide range of cost estimates and clinical event rates. CONCLUSION: Despite its greater initial cost, DOT is a more cost-effective strategy than SAT because it achieves a higher cure rate after initial therapy, and thereby decreases treatment costs associated with failure of therapy and acquired drug resistance. This cost-effectiveness analysis supports the widespread implementation of DOT. PMID- 9228361 TI - Atherosclerotic aortic plaque detected by transesophageal echocardiography: its significance and limitation as a marker for coronary artery disease in the elderly. AB - OBJECTIVES: To elucidate whether atherosclerotic aortic plaque detected by transesophageal echocardiography can be a clinically useful marker for coronary artery disease in the elderly. BACKGROUND: Atherosclerotic aortic plaque detected by transesophageal echocardiography has been reported to be a marker for coronary artery disease. Its significance may be important particularly in the elderly population, although to our knowledge, there are no data yet available. METHODS: We performed transesophageal echocardiography on 84 patients who had previously undergone coronary arteriography. The criteria used to diagnose atherosclerotic plaque on transesophageal echocardiography were the presence of focally or linearly increased echodensity of the aortic intima with lumen irregularity and thickening or ulceration. RESULTS: Significant coronary artery disease (> or = 50% stenosis) was detected in at least one major coronary artery in 27 of the 84 patients. Aortic plaques were detected by transesophageal echocardiography in 25 of the 27 patients (93%) with coronary artery disease and in 30 of 57 patients (53%) without coronary disease (p<0.001). Among 24 patients 70 years or older, aortic plaques were present in 13 of 14 (93%) patients with coronary artery disease and 9 of 10 patients (90%) without coronary disease. Among 60 patients younger than 70 years, aortic plaques were present in 12 of 13 patients (92%) with coronary artery disease and 21 of 47 patients (45%) without coronary disease (p<0.01). The independent association between coronary artery disease and the presence of aortic plaque, age, gender, and other coronary risk factors was examined by multiple logistic regression analysis. In patients 70 years or older, the presence of aortic plaque failed to be a predictor of significant coronary artery disease, although it was indeed a strong predictor of coronary artery disease in patients younger than 70 years (p<0.05). CONCLUSIONS: In elderly patients, atherosclerotic aortic plaque detected by transesophageal echocardiography is not useful in predicting significant coronary artery disease. It is useful only in a relatively younger population. PMID- 9228362 TI - Factors associated with variations in pulmonary function among elderly Japanese American men. AB - OBJECTIVE: To identify lifestyle, anthropometric, biochemical, and clinical characteristics associated with pulmonary function in elderly men. DESIGN: Cross sectional population-based study. PARTICIPANTS: Japanese-American men (n=3,111) aged 71 to 93 years, who completed spirometry at the fourth examination of the Honolulu Heart Program (1991 to 1993). METHODS: Pulmonary function measurements (FEV1 and FVC) were obtained using American Thoracic Society guidelines. Potential factors associated with pulmonary function were examined using Pearson correlation coefficients and general linear models. Age- and height-adjusted mean levels of FEV1 and FVC were compared across quintiles of continuous variables and by status of prevalent disease and medication or vitamin use. Stepwise multiple linear regression was used to identify factors independently associated with pulmonary function overall and among never smokers. RESULTS: A number of correlates of pulmonary function were initially identified. In multivariate analyses, age, current smoking, pack-years of smoking, emphysema, asthma, wheezing without colds, subscapular skinfold thickness, ECG abnormality, heart rate, WBC count, and eosinophil count were all negatively and independently associated with FEV1, while height, grip strength, physical activity, and mean corpuscular hemoglobin concentration were positively associated. With a few exceptions, similar relations were observed with FVC and among never-smokers. CONCLUSION: Cigarette smoking, respiratory symptoms and disease, and several cardiovascular risk factors were independently associated with pulmonary function in elderly Japanese-American men. In most cases, inadequate control for smoking does not appear to account for these associations. Results suggest that a number of factors that are correlates of FEV1 and FVC in younger age groups are also associated with pulmonary function beyond the age of 70 years. PMID- 9228363 TI - Evaluation of a new treadmill exercise protocol. AB - STUDY OBJECTIVES: To confirm that a newly drafted treadmill exercise protocol designed on a theoretical basis to span a range of 0 to 200 W with approximately 25-W increments by alteration of either speed or grade from one stage to the next should correspond to a standard bicycle protocol consisting of 25-W steps. DESIGN: Randomized, crossover study to compare both exercise test modes. STUDY PARTICIPANTS: Twenty-one consecutive healthy volunteers. INTERVENTIONS: Subjects underwent both exercise tests until either exhaustion or completion of the respective protocol, and cardiopulmonary exercise parameters were assessed during either of them. For comparison, correlation coefficients (r) were calculated. RESULTS: Exercise tolerance time was 9% higher on the treadmill (p<0.05). Ten subjects completed the bicycle program, whereas 18 subjects did so on the treadmill. With both protocols, there were comparably linear increases in heart rate (r=0.885), oxygen uptake (r=0.925), oxygen uptake per body weight (r=0.944), carbon dioxide output (r=0.937), and minute ventilation (r=0.914). For the 2-min stage duration, a plateau in oxygen uptake was achieved with neither protocol. The ventilatory equivalent for oxygen, which is not linear, showed its minimum at comparable workloads, at the point of surpassing the anaerobic threshold. Correlation of oxygen pulse was fair (r=0.896). CONCLUSIONS: There was an excellent correlation of the parameters with respect to both measured values at identical workloads and slopes of both protocols, thus enabling comparability of treadmill and bicycle ergometry. Due to its practical handling, the new protocol may facilitate acceptance, especially when used for elderly or disabled patients. PMID- 9228364 TI - Measurements of PEEP-induced changes in lung volume: evaluation of a laser monitor. AB - STUDY OBJECTIVE: To evaluate a newly developed laser monitor in the measurement of positive end-expiratory pressure (PEEP)-induced changes in end-expiratory lung volume (EELV) in spontaneously breathing subjects. DESIGN: An open comparison between two different methods to assess breathing parameters. SETTING: A respiratory research unit. PARTICIPANTS: Six spontaneously breathing, healthy volunteers. INTERVENTIONS: Stepwise increases of PEEP from 0 to 0.8 to 1.6 kPa during spontaneous breathing; repeated validation of the laser monitor in each subject. MEASUREMENTS AND RESULTS: Pressure and flow were recorded at the airway opening. Abdominal wall displacement (AWD) measured by the laser sensor was recorded simultaneously. The time lag between the volume and the laser signals during baseline was 0.068+/-0.052 s and during maximal PEEP, 0.108+/-0.093 s. There was no baseline drift in either the PEEP or the AWD signals. Mean EELV decreased by 290 mL to 1,157 mL after the PEEP valve was removed. Within each individual, the ratio between EELV and AWD showed only small variations. Measurements of tidal volume (VT) and AWD showed good agreement at all PEEP levels. Mean VT decreased in all but one subject during PEEP. With the increase in PEEP, the end-expiratory abdominal baseline increased linearly over a large range of end-expiratory pressures with a flattening of the curve at high PEEP levels in all subjects. CONCLUSIONS: The laser monitor is sufficiently accurate for measuring PEEP-induced changes in EELV during spontaneous breathing in healthy adult subjects. Monitors incorporating multiple laser sensors may have considerable clinical promise. PMID- 9228365 TI - Maximal expiratory pressures in spinal cord injury using two mouthpieces. AB - STUDY OBJECTIVE: A technique for assessing expiratory muscle strength is the measurement of maximal expiratory pressure (PEmax). Previous studies have shown that a tube-style mouthpiece yields greater PEmax values than a flange-style mouthpiece because the latter technique is limited by the strength of the buccal muscles. In individuals with weak muscles of exhalation, this limitation may not apply because the strength of their buccal muscles may exceed that of the respiratory muscles. DESIGN: A tube-style mouthpiece and flange-style mouthpiece were used to measure PEmax. The order of the mouthpiece used in testing was alternated between subjects and the greatest values obtained after three efforts were compared. SETTING: Department of Veterans Affairs Medical Center. PARTICIPANTS: Fifty subjects with chronic spinal cord injury without acute medical illnesses recruited from veterans and the community. RESULTS: The mean difference between PEmax(tube) and PEmax(flange) was 20.7+/-26.4 cm H2O (p = 0.0001). Differences were negligible in those with the weakest muscles of exhalation but were substantial even in some quadriplegic subjects. CONCLUSION: Even in individuals with neuromuscular disorders, errors in assessment of expiratory strength occur when a flange-style mouthpiece is used, and we recommend that this technique be abandoned in the measurement of PEmax. PMID- 9228366 TI - Noninvasive ventilatory support after lung resectional surgery. AB - STUDY OBJECTIVES: To investigate the short-term effects of noninvasive ventilatory support (NIVS) on pulmonary gas exchange, ventilatory pattern, systemic hemodynamics, and pleural air leaks in patients submitted to elective lung resection. DESIGN: Prospective, randomized, parallel, and controlled investigation. SETTING: Thoracic Surgery Unit, Hospital Universitari Son Dureta, Palma Mallorca, Spain. PATIENTS: Nineteen patients electively submitted to lung resection because of varied clinical reasons. INTERVENTIONS: Medical therapy was standardized for all patients. Ten subjects received NIVS with a nasal ventilatory support system (BiPAP) during 1 h (study group). The remaining nine individuals constituted the control group. MEASUREMENTS AND RESULTS: Arterial blood gases, ventilatory pattern, systemic hemodynamics, and pleural air leaks were measured. Before surgery, there were no significant clinical or functional differences between groups. After surgery, and compared with preoperative measures, PaO2 decreased significantly (p<0.01) and to the same extent both in the study group (85.7+/-2.8 to 68.0+/-2.7 mm Hg) and the control group (83.6+/ 2.5 to 67.3+/-2.6 mm Hg). In the study group, NIVS increased PaO2 (to 76.7+/-3.0 mm Hg; p<0.05) and decreased alveolar to arterial oxygen pressure gradient (P[A a]O2) (27.2+/-2.7 to 17.6+/-2.3 mm Hg; p<0.05). This latter effect was still present 1 h after withdrawing NIVS. By contrast, PaO2 and P(A-a)O2 remained unchanged in the control group throughout the study. PaCO2, the ventilatory pattern, and systemic hemodynamics did not change significantly throughout the study in any group. Importantly, NIVS did not increase dead space to tidal volume ratio or worsen pleural air leaks. CONCLUSIONS: Short-term NIVS with a ventilatory support system improves the efficiency of the lung as a gas exchanger without noticeable nondesired side effects in patients submitted to lung resectional surgery. PMID- 9228367 TI - Coronary artery disease in patients undergoing lung volume reduction surgery for emphysema. AB - OBJECTIVES: Most patients with severe pulmonary emphysema referred for lung volume reduction surgery (LVRS) have a long-standing history of cigarette smoking. Coronary artery disease (CAD) predisposes to perioperative cardiac complications. Since symptoms and signs of myocardial ischemia are often absent in patients with severe ventilatory impairment even during exercise, we investigated the prevalence of CAD in candidates for LVRS by angiography. DESIGN: We prospectively studied the prevalence of CAD by angiography and assessed the CAD risk factor profile in 41 candidates for LVRS (26 men, 15 women; mean age, 66+/-6.8 years; range, 52 to 76 years), who had no current symptoms or a history of myocardial ischemia. RESULTS: In six patients (15%), asymptomatic but significant coronary lesions (> 70% stenosis) were detected. In five patients, these findings altered the clinical management. Patients with CAD had significant higher cholesterol levels, tended to have smoked more, and had more often additional vascular risk factors. CONCLUSIONS: We found a high prevalence of angiographically significant but clinically silent CAD in this particular population of heavy smokers with advanced emphysema. PMID- 9228368 TI - Foreign body aspiration into the lower airway in Chinese adults. AB - OBJECTIVES: Foreign body aspiration into the lower airway in adults is uncommon. We designed this study to investigate the clinical presentations, precipitating factors, management choice, and complications of foreign body aspiration in Chinese adults. PATIENTS AND METHODS: We analyzed 43 consecutive adult patients with foreign body aspiration between February 1980 and December 1995 from the medical record registry and cross index system of a tertiary medical center. RESULTS: The most common symptoms are chronic cough, hemoptsis, fever, and dyspnea. Only three patients (7%) presented with choking. Chest radiograph demonstrated the foreign body in nine cases (21%). The most common foreign body was bone fragments (21/43, 49%). Lodgment is more common in the right, especially the right intermediate bronchus and basal bronchus. Three patients were also diagnosed as having lung cancers. Precipitating factors include CNS dysfunction, facial trauma, intubation, dental procedure, and underlying pulmonary diseases. Flexible fiberoptic bronchoscopy removed the foreign body in 25 cases (58%) during the first attempt and 32 cases (74%) in total. Complications include obstructive pneumonitis (including one case of actinomycosis infection), atelectasis, bronchiectasis, lung abscess, and lung torsion (two cases). CONCLUSION: The nature of foreign body in Chinese adults was different from the Western adults. The initial clues to foreign body aspiration in adults are usually obscure or indirect. We suggest flexible fiberoptic bronchoscopy as the first-line approach. Follow-up bronchoscopy and chest radiograph are recommended to detect chronic complications or coexisting lung cancer. PMID- 9228369 TI - Implantation of Accuflex and Strecker stents in malignant bronchial stenoses by flexible bronchoscopy. AB - Silicone and metal stents are available for the treatment of malignant bronchial stenoses. This project sought to compare the self-expanding nitinol Accuflex stent (Boston Scientific Corp; Watertown, Mass) with the passively expandable tantalum Strecker stent (Boston Scientific Corp; Watertown, Mass), both implanted by flexible bronchoscopy under local anesthesia and sedation. In 51 patients with malignant bronchial stenosis, 14 nitinol and 51 tantalum stents were used and stenoses of 75 to 100% were treated. The intervention was successful in all but one patient; a mean patency of 93% was achieved. In the follow-up period, the probability of survival was significantly lower in patients with total bronchus occlusion than in patients with stenotic alterations (44 vs 109 days; p<0.05). In 10 patients, lung function analysis after stent implantation revealed a significant increase in PaO2 (65 vs 71 mm Hg; p<0.01), inspiratory vital capacity (2.5 vs 2.7 L; p<0.05), and FEV1 (1.8 vs 2.0 L; p<0.05). Mucus retention was the main (39%) adverse factor in the early phase after stent implantation, whereas tumor penetration became the most frequent problem (67%) in the later phase. Recanalizing interventions were necessary in 18% of the cases in which tumor penetration occurred. Stent distortion occurred in 12 patients with Strecker and in none with Accuflex stents. In comparison to the Strecker stent, the self expanding Accuflex stent is preferable owing to its excellent flexibility and faster delivery system. Both types of stents could be sufficiently deployed within the lesion and allowed for highly precise positioning. Furthermore, no general anesthesia was required. The fiberbronchoscopic mode of implantation under sedation is very efficient even for tumor patients with severe impairment of their physical and respiratory condition. PMID- 9228370 TI - Influence of autonomic neuropathy of different severities on the hypercapnic drive to breathing in diabetic patients. AB - To investigate the effects of the autonomic nervous system on control of breathing, the neuromuscular (mouth occlusion pressure at 0.1 s after onset of inspiration [P0.1]) and ventilatory (minute ventilation [VE]) response to progressive hyperoxic hypercapnia was assessed in diabetic patients with autonomic dysfunction of different severity. Eighteen diabetics with autonomic neuropathy, nine with parasympathetic damage (DANp), and nine with parasympathetic and sympathetic damage (DANp+s), as indicated by marked postural hypotension, low increment of diastolic BP during sustained handgrip, and lowest resting catecholamine plasma levels, were studied together with a group of 10 diabetic patients without autonomic neuropathy (D) and a group of 10 normal subjects (C). All subjects had pulmonary function tests, including maximal voluntary ventilation and diffusion of carbon monoxide, measurements of respiratory muscle strength as maximal inspiratory mouth pressure (MIP) and maximal expiratory mouth pressure (MEP), and a CO2 rebreathing test (Read's method). Although in the normal range, lung volumes and FEV1 and forced expiratory flows were lower in the DANp and DANp+s groups than in the D and C groups, MIP and MEP were similar among C and diabetic groups, as well as resting P0.1, VE, tidal volume (VT), and respiratory rate (RR). The slope of the linear relationship between P0.1 and end-tidal PCO2 (PETCO2) was higher in DANp+s (0.63+/-0.07 cm H2O/mm Hg) than in C (0.45+/-0.06 cm H2O/mm Hg; p<0.05) and three times greater in DANp+s than in D (0.26+/-0.03 cm H2O/mm Hg; p<0.001) and DANp (0.24+/-0.03 cm H2O/mm Hg; p<0.001), who in turn showed a lower deltaP0.1/deltaPETCO2 than C. The VE increase with increasing PETCO2 was greater in DANp+s (3.70+/-0.85 L/min/mm Hg) than in DANp (2.13+/-0.20 L/min/mm Hg; p<0.05) and D (2.37+/-0.40 L/min/mm Hg; p=0.07), but not significantly higher from that of C (3.17+/-0.36 L/min/mm Hg). No differences were found for deltaVT/deltaPETCO2 among the groups, whereas the deltaRR/deltaPETCO2 relationship was steeper in DANp+s than in DANp (p<0.05) and D (p=0.055). These data reflect a depressed CO2 response both in D and DANp. The presumable decrease of the sympathetic nerve traffic in DANp+s appears to reverse this abnormality. DANp+s, however, exhibit an enhanced CO2 neuromuscular response even in respect to C, suggesting that the sympathetic nervous system might modulate the output of the respiratory centers to hypercapnic stimulus. PMID- 9228371 TI - Hypoxic ventilatory response and breathlessness following hypocapnic and isocapnic hyperventilation. AB - STUDY OBJECTIVES: To investigate the etiology of posthyperventilation (post-HV) hypoxemia following voluntary hyperventilation (VHV), we examined the effects of hypocapnic (hypo-CO2) and isocapnic (iso-CO2) VHV on the hypoxic ventilatory response (O2-response) and on the sensation of breathlessness during the O2 response. METHODS: O2-responses and visual analog scale (VAS) scores for estimating breathlessness in 10 normal subjects during the O2-response under iso CO2 conditions and under hypo-CO2 conditions immediately following voluntary maximal HV of 3 min duration were examined. RESULTS: Although there was no significant difference in the post-HV ventilation levels following hypo-CO2 vs iso-CO2 VHV, the VAS scores at the start of the O2-response following hypo-CO2 VHV (30.2+/-24.2 mm) were significantly higher (p<0.05) than the VAS scores at the start of the O2-response following iso-CO2 VHV (13.7+/-8.4 mm). However, VHV did not have a significant effect on the O2-response at 2 min after the VHV when the arterial O2 saturation (SaO2) was below 90%. The nonsteady-state hypo-CO2 induced by VHV greatly attenuated the O2-response below 90% SaO2 and VAS scores at 70% SaO2. CONCLUSIONS: Elevated VAS scores immediately following the hypo-CO2 VHV, which might be independent of actual breathing levels, and the attenuation of the O2-response following the hypo-CO2 VHV were not due to input from lung and chest wall mechanoreceptors induced by the hyperpnea itself, but rather to the hypo-CO2 induced by hyperpnea. PMID- 9228373 TI - A rapid qualitative assay to detect circulating endotoxin can predict the development of multiorgan dysfunction. AB - OBJECTIVE: To determine whether a rapid qualitative assay for the detection of circulating endotoxin (SimpliRED Endotoxin Test [SRE]; AGEN, Inc; Brisbane, Australia) can predict the occurrence of multiorgan dysfunction and hospital mortality. To compare the SRE to the limulus amebocyte lysate (LAL) assay as a predictor of clinical outcomes. DESIGN: Prospective, blinded, single-center study. SETTING: Medical ICU of Barnes-Jewish Hospital, St. Louis, a university affiliated teaching hospital. PATIENTS: Included in the study were 265 adult patients requiring medical ICU admission. INTERVENTIONS: Daily collection of blood samples. MEASUREMENTS AND RESULTS: Daily detection for the presence of endotoxin in blood during intensive care and assessment for the development of multiorgan dysfunction (ie, an organ system failure index >2) or death. On ICU day 1, 55 (20.8%) patients had circulating endotoxin detected by the SRE. On ICU day 2, 29 of the 143 (20.3%) patients remaining in the ICU had a positive SRE. The development of multiorgan dysfunction was significantly greater among SRE positive patients (44.8%) compared to SRE-negative patients (21.9%) on ICU day 2 (p=0.013). Multiple logistic regression analysis identified a positive SRE on ICU day 2 (adjusted odds ratio, 4.1; 95% confidence interval, 2.5 to 6.8; p=0.006) as being independently associated with the development of multiorgan dysfunction. A positive SRE test was not predictive of hospital mortality. Direct quantitative measurement of the concentration of circulating endotoxin using the LAL assay was not associated with either the development of multiorgan dysfunction or hospital mortality (p>0.1). CONCLUSIONS: Our preliminary data suggest that a bedside assay to qualitatively detect circulating endotoxin is predictive of the development of multiorgan dysfunction among patients admitted to a medical ICU. The rapid detection of circulating endotoxin could be useful for stratifying patients into various risk categories for the development of multiorgan dysfunction. PMID- 9228372 TI - A trial of antioxidants N-acetylcysteine and procysteine in ARDS. The Antioxidant in ARDS Study Group. AB - OBJECTIVE: To determine the levels of glutathione and cysteine in patients with ARDS and examine the effect of treatment with N-acetylcysteine (NAC) and L-2 oxothiazolidine-4-carboxylate (Procysteine; Clintec Technologies Inc; Chicago [OTZ]) on these levels and on common physiologic abnormalities, and organ dysfunction associated with ARDS. DESIGN: Randomized, double-blind, placebo controlled, prospective clinical trial. SETTING: ICUs in five clinical centers in the United States and Canada. PATIENTS: Patients meeting a predetermined definition of ARDS and requiring mechanical ventilation. INTERVENTION: Standard care for ARDS and I.V. infusion, every 8 h for 10 days, of one of the following: NAC (70 mg/kg, n=14), OTZ (63 mg/kg, n=17), or placebo (n=15). MAIN RESULTS: Both antioxidants effectively repleted RBC glutathione gradually over the 10-day treatment period (47% and 49% increases from baseline values for NAC and OTZ, respectively). There was no difference in mortality among groups (placebo, 40%; NAC, 36%; OTZ, 35%). However, the number of days of acute lung injury was decreased and there was also a significant increase in cardiac index in both treatment groups (NAC/OTZ [+]14%; placebo [-]6%). CONCLUSIONS: Our findings suggest that repletion of glutathione may safely be accomplished with NAC or OTZ in patients with acute lung injury/ARDS. Such treatment may shorten the duration of acute lung injury, but larger studies are needed to confirm this. PMID- 9228374 TI - Severe paroxysmal sinus bradycardia associated with high-frequency oscillatory ventilation. AB - OBJECTIVE: To determine the incidence of and risk factors for unexplained paroxysmal bradycardia in children treated with high-frequency oscillatory ventilation (HFOV). DESIGN: A nested case-control study. SETTING: A university affiliated children's hospital. SUBJECTS: All children treated with HFOV for at least 3 days during a 2-year period and a randomly chosen comparison group of 50 children treated with only conventional mechanical ventilation (CMV) for at least 3 days during the same time period. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: Unexplained paroxysmal sinus bradycardia occurred in six children (12%) receiving HFOV, and was significantly more common than in children treated with CMV (0%). The bradycardic events occurred after the lung disease started to improve, and the mean airway pressure (mPaw) at the time of the bradycardias was significantly decreased from the child's maximal mPaw. The bradycardic events were effectively treated acutely with manual ventilation or atropine sulfate, and resolved completely after the patient was changed to a regimen of CMV. CONCLUSION: Unexplained paroxysmal bradycardia associated with HFOV in children is not uncommon. It completely resolves with conversion to CMV and may be related to overdistention of alveoli as compliance improves. PMID- 9228375 TI - Effect of failed extubation on the outcome of mechanical ventilation. AB - OBJECTIVE: To examine medical outcomes associated with reintubation for extubation failure after discontinuation of mechanical ventilation. DESIGN: Prospective cohort study of consecutive intubated medical ICU patients who underwent a trial of extubation at a tertiary-care teaching hospital. The failed extubation group consisted of all patients reintubated within 72 h or within 7 days (if continuous ICU care had been required) of extubation. All others were considered to be successfully extubated. Study end points included hospital death vs survival, the number of days spent in the ICU and in the hospital after the onset of mechanical ventilation, the likelihood of requiring > or = 7 or > or = 14 days of ICU care after extubation, and the need for transfer to either a long term care or rehabilitation facility among the survivors. RESULTS: Of 289 intubated patients, 247 (85%) were successfully extubated, and 42 (15%) required reintubation for failed extubation (time to reintubation 1.5+/-0.2 days). Reintubation for extubation failure resulted in 12 additional days of mechanical ventilation. When compared with successfully extubated patients, reintubated patients were more likely to die in the hospital (43% vs 12%; p<0.0001), spend more time in the ICU (21.2+/-2.8 days vs 4.5+/-0.6 days; p<0.001) and in the hospital (30.5+/-3.3 days vs 16.3+/-1.2 days; p<0.001) after extubation, and require transfer to a long-term care or rehabilitation facility (38% vs 21%; p<0.05). Using multiple logistic regression, extubation failure was an independent predictor for death and the need for transfer to a long-term care facility. Compared with those successfully extubated, patients who failed extubation were seven times (p<0.0001) more likely to die, 31 times (p<0.0001) more likely to spend > or = 14 days in the ICU after extubation, and six times (p<0.001) more likely to need transfer to a long-term care or rehabilitation facility if they survived. CONCLUSION: After adjusting for severity of illness and comorbid conditions, extubation failure had a significant independent association with increased risk for death, prolonged ICU stay, and transfer to a long-term care or rehabilitation facility. Extubation failure may serve as an additional independent marker of severity of illness. Alternatively, poor outcomes may be etiologically related to extubation failure. If the latter proves to be the case, identifying patients at risk for poor outcomes from extubation failure and instituting alternative care practices may reduce mortality, duration of ICU stay, and need for transfer to a long-term care facility. PMID- 9228377 TI - Therapeutic rigid bronchoscopy allows level of care changes in patients with acute respiratory failure from central airways obstruction. AB - OBJECTIVE: To determine whether emergency rigid bronchoscopic intervention, including Nd-YAG laser resection or stenting, immediately affected the need for continued mechanical ventilation or intensive care level of support in critically ill patients with acute respiratory failure from malignant or benign central airways obstruction. DESIGN: Retrospective review of medical records of all patients with acute respiratory failure and malignant or benign tracheobronchial obstruction necessitating intubation, mechanical ventilation, or hospitalization in the ICU prior to referral for therapeutic bronchoscopy. SETTING: University of California San Diego, a tertiary care institution specialized in airway management. PATIENTS: Medical records of 32 patients with malignant or benign central airways obstruction requiring admission to the ICU prior to rigid bronchoscopic intervention between January 1994 and April 1996. INTERVENTIONS: Emergent rigid bronchoscopy with dilatation, Nd-YAG laser resection, or silicone stent insertion performed in the operating room under general anesthesia. RESULTS: Thirty-two patients with central airways obstruction requiring emergent hospitalization in the ICU were referred for therapeutic rigid bronchoscopy. Airway strictures were caused by benign disease in 18 patients, and by primary bronchogenic lung cancer in 14. Of the 19 patients who were mechanically ventilated, bronchoscopic intervention allowed immediate discontinuation of mechanical ventilation in 10 (52.6%). Twenty-five patients had indwelling artificial airways (12 endotracheal tubes, 13 tracheotomy tubes). Two, however, were considered tracheotomy-dependent because of neuromuscular disease. Of the remaining 23 patients, immediate extubation or decannulation was possible in seven (30.4%). Of seven patients with no indwelling airway, five (71.4%) were immediately transferred to a lower level of care after intervention. Of the 32 total patients, 20 (62.5%) were immediately transferred to a lower level of care immediately after intervention. CONCLUSIONS: Emergency laser resection or stent insertion can favorably affect health-care utilization in patients with acute respiratory distress from central airways obstruction. Treatment may be lifesaving and allows successful withdrawal from mechanical ventilation, hospitalization in a lower level of care environment, relief of symptoms, and extended survival in critically ill patients. In patients with regionally advanced cancer, the palliative nature of this procedure postpones death by respiratory distress and may prompt consideration for institution of conservative comfort measures to reduce patient suffering. PMID- 9228376 TI - Early vs conventional extubation after cardiac surgery with cardiopulmonary bypass. AB - OBJECTIVES: Sedation and ventilation overnight after cardiac surgery is common practice. However, early extubation may be feasible with no increase in postoperative complications. This study examines (1) if early extubation is possible in a significant number of patients, (2) if it reduces ICU stay, and (3) if this practice increases postoperative complications. DESIGN: Prospective, controlled, randomized clinical trial. PATIENTS AND METHODS: We randomized 404 consecutive patients to early extubation (7 to 11 h postoperatively) (group A, 201 patients) or conventional extubation (between 8 and 12 AM the following day) (group B, 203 patients). Variables included type and severity of the disease, surgical risk, type of operation, operative incidences, postoperative complications, duration of mechanical ventilation, intubation and ICU stay, bleeding, reoperation, vasoactive drugs, and mortality. RESULTS: Groups were comparable. Extubation within the preestablished time was successful in 60.2% of patients in group A and 74.4% in group B. Median ICU stay was 27 h in group A and 44 h in group B (p=0.008). Discharge from ICU within the first 24 h postoperatively was 44.3% in group A and 30.5% in group B (p=0.006). There was no significant difference in complications between groups. Successfully extubated patients in group A had more reintubation and prolonged ventilation than in group B. CONCLUSIONS: (1) Sixty percent of our patients were extubated within 11 h of operation. (2) As a result, the length of stay in ICU was reduced and the percentage of patients discharged within 24 h was increased. (3) There was no increase in clinically important postoperative complications. PMID- 9228378 TI - Noninvasive positive pressure ventilation and not oxygen may prevent overt ventilatory failure in patients with chest wall diseases. AB - Some patients with chest wall diseases (CWD) without respiratory failure manifest important alterations in nocturnal gas exchange, as a previous stage to the future development of daytime respiratory failure. The purpose of this study was to evaluate the efficacy of nasal intermittent positive pressure ventilation (NIPPV) during sleep in a group of obese patients and in another group with restrictive thoracic diseases (RTD), comparing the results with those obtained from conventional nocturnal oxygen therapy. From a total of 42 patients with CWD free of daytime respiratory failure, 27 (64%) were considered nocturnal oxygen desaturators without sleep apnea and were included in the study. The study protocol was completed by 21 of these patients. After 2 weeks of treatment, symptoms of dyspnea, morning headaches, and morning obnubilation improved significantly (p<0.05) in both groups of patients after NIPPV but not with oxygen. Baseline daytime PaO2 was 68+/-7 mm Hg in the obese group of patients and 73+/-11 mm Hg in the RTD group. It improved significantly with NIPPV to 73+/-5 mm Hg in obese patients (p<0.05) and to 77+/-12 mm Hg in the RTD group (p<0.05) but did not change with oxygen (68+/-8 mm Hg in the obese group and 73+/-12 mm Hg in the RTD group). Both treatments improved oxygen saturation during sleep, but oxygenation tends to be higher with oxygen than with NIPPV. Only NIPPV was able to normalize the baseline nocturnal alveolar hypoventilation. From the 21 patients treated, 19 decided to continue with long-term NIPPV, one with oxygen, and one refused treatment. We conclude that in patients with CWD who manifest nighttime oxygen desaturation and hypoventilation, early initiation of NIPPV is preferable to supplemental oxygen. Our results also suggest that NIPPV initiated before overt ventilatory failure could prevent its onset. PMID- 9228379 TI - Effect of exposure of guinea pigs to cigarette smoke on elastolytic activity of pulmonary macrophages. AB - STUDY OBJECTIVE: To determine the effect of exposure to cigarette smoke on the elastolytic activity of guinea pigs' alveolar macrophages (AMs), and to compare elastolytic activity of AMs obtained by BAL with that of lung macrophages (LMs) obtained from minced lung tissue. METHODS: AMs were obtained by BAL from seven adult guinea pigs exposed to cigarette smoke for 5 d/wk during 6 weeks, as well as from age-matched control guinea pigs. From each animal, one lung was used to obtain LMs by mincing and teasing the lung, followed by enzymatic digestion and isolation of mononuclear cells by Hypaque-Ficoll separation. The other lung was inflated and fixed to quantitate emphysema by the destructive index (DI). Elastolytic activity (microgram of elastin degraded by 10(6) macrophages) was determined at 24, 48, and 72 h, by culturing AMs and LMs (1 x 10(6) cells in 1 mL of medium) in 3H-elastin-coated wells. RESULTS: In animals exposed to cigarette smoke, the total number of BAL cells (8.6+/-2.1 x 10(6)) and DI (21.8+/-8.1) were significantly higher than in nonexposed animals (6.4+/-1.8 x 10(6), p<0.05 for cells, and 12.1+/-4.1, p<0.01 for DI). Elastolytic activity of AMs from smoke exposed guinea pigs was significantly higher at 24, 48, and 72 h than elastolytic activity of AMs from control animals (19.0+/-9.4 vs 10.0+/-5.3, p<0.05 at 72 h). Likewise, elastolytic activity of LMs was significantly higher in exposed than nonexposed guinea pigs (11.8+/-7.7 vs 7.4+/-5.0 at 72 h, p<0.05). Elastolytic activity of LMs was not significantly different from elastolytic activity of AMs, both in exposed guinea pigs (11.8+/-7.7 vs 19.0+/-9.4 at 72 h) and nonexposed animals (7.4+/-5.0 vs 10.0+/-5.3 at 72 h). CONCLUSIONS: These results indicate that elastolytic activity of both AMs and LMs of guinea pigs increases significantly after exposure to cigarette smoke and that AMs and LMs have similar elastolytic activities. PMID- 9228380 TI - Neurosarcoidosis: a personal perspective based on the study of 37 patients. AB - Clinically apparent involvement of the nervous system occurs in a relatively small number of patients with sarcoidosis. The diagnosis of neurosarcoidosis is often difficult and particularly so in patients who lack either pulmonary or systemic manifestations of sarcoidosis. Furthermore, clinical features of neurosarcoidosis are extremely variable. In this series of 37 patients, seen during the last 30 years, cranial nerve palsies occurred in 52%, polyneuritis or polyneuropathy in 24%, meningeal involvement in 24%, muscle disease in 8%, and Guillain-Barre syndrome in 5% of the patients. Other presentations included seizures, brain mass, pituitary/hypothalamic syndrome, and memory loss associated with confusion. The chest radiograph was abnormal in 8 of every 10 patients with neurosarcoidosis. In 18 (85%) of 21 patients, gallium uptake was consistent with the diagnosis of active sarcoidosis. Serum angiotensin-converting enzyme levels were raised in about half of the patients. Cerebrospinal fluid features, including lymphocyte pleocytosis, raised protein levels, and decreased glucose concentration, were of little help. MRI with gadolinium enhancement was the most sensitive diagnostic tool, particularly in patients with meningeal involvement. The ultimate arbiter of the diagnosis of neurosarcoidosis, the presence of noncaseating granulomas in the involved tissue, was not always available. Although corticosteroids are the mainstay of therapy, in this series, 12 patients received chloroquine or hydroxychloroquine. Prognosis of chronic neurosarcoidosis is poor. Six (18%) of 37 patients died of complications related to sarcoidosis. PMID- 9228381 TI - IIB or not IIB: the current question in staging non-small cell lung cancer. AB - It has been suggested that T3/N0-1/M0 non-small cell lung cancer should be classified as stage IIB rather than IIIA. This is the result of a widespread perception that the survival of patients with T3/N0-1 lung cancers greatly exceeds that of patients with stage IIIA (N2) lung cancers. This perception is based primarily on the survival of T3/N0-1 patients who have chest wall involvement. However, the T3 classification also includes tumors that involve mediastinal structures, the main stem bronchus <2 cm from the carina, and the brachial plexus as seen in Pancoast tumors. Survival for each of these T3 categories is examined in this articles and found to be somewhat different. The available data show that patients with T3/N0-1 tumors involving the chest wall have a good prognosis after resection, whereas patients with central T3/N0-1 tumors (mediastinal or main stem bronchial involvement) have a prognosis similar to that of patients with resected IIIA (N2) tumors. If a new classification of T3/N0-1 tumors as stage IIB is to be adopted, it will be important for future studies to document which type of T3 tumor is being discussed. PMID- 9228382 TI - Sepsis: a new hypothesis for pathogenesis of the disease process. PMID- 9228383 TI - Unstable angina: are we able to recognize high-risk patients? AB - It is difficult to identify characteristics of patients with unstable angina that are predictive of a high likelihood of developing clinical events. However, several features have been recognized. Patients with a clinical history of previous stable exertional angina symptoms who began to experience rest pain appear to be at risk and tend to have more extensively underlying coronary disease. When the ischemic episodes are accompanied by rates, a new or worsening mitral regurgitation murmur, or hypotension, there is a high likelihood of significant coronary artery disease and one should triage these patients to early cardiac catheterization and prompt revascularization. An angiographic feature that carries a high risk is a lesion in the proximal left anterior descending or in the left main coronary artery. Certain typical ECG patterns are very suggestive for a critical narrowing in these coronary arteries. If chest pain and ST-segment changes recur on vigorous medical management, early invasive evaluation should be strongly considered. Even so, the left ventricular function is very important prognostically. According to serologic tests, the level of C reactive protein and serum amyloid A protein suggesting that there may be active inflammation predicts an early poor outcome. However, these serologic abnormalities do not have much clinical value. An increased platelet activation and a reduced fibrinolytic capacity play a role in the pathogenesis of unstable angina, but thrombolytic therapy does not improve the prognosis in patients with unstable angina. PMID- 9228385 TI - Antitumor activity of a monoclonal antibody directed against gastrin-releasing peptide in patients with small cell lung cancer. AB - BACKGROUND: Small cell lung cancer (SCLC) cells express and secrete gastrin releasing peptide (GRP) which binds to receptors and stimulates growth of these cells. A murine monoclonal antibody, 2A11, which binds GRP with high affinity, decreased growth of SCLC cells in vitro and in athymic nude mice. A phase 1 trial and pharmacokinetic modeling in patients with lung cancer has defined the phase 2 dose of 2A11 but the antitumor activity in patients is unknown. METHODS: Thirteen patients with previously treated SCLC received 2A11 at 250 mg/m2 over 1 h three times per week for 4 weeks. Serum GRP, urine GRP, serum levels of 2A11, and human antimouse antibodies (HAMA) were determined. RESULTS: One of 12 (8%; 95% confidence interval, 0 to 38%) evaluable patients had complete resolution of radiographically detectable tumor lasting 4 months. Four patients (33%) had stable disease. No toxic reactions were observed. The pretreatment serum GRP level of the responding patient was 3.1 fmol/mL and the median of nine nonresponding patients was 7.3 fmol/mL (range, <1.0 to 29.0). The mean trough serum 2A11 level was 49+/-18 microg/mL in the responding patient and 32 to 487 mg/mL (median, 117) in 10 nonresponding patients. HAMA did not increase during 2A11 administration in any patient. CONCLUSIONS: Interruption of the GRP autocrine growth factor loop with 2A11 results in clinical antitumor activity in a minority of patients with previously treated SCLC. Further evaluation of the antitumor effects of 2A11 is warranted to define characteristics associated with response to 2A11. PMID- 9228384 TI - Underutilization of transbronchial needle aspiration: experiences of current pulmonary fellows. AB - Transbronchial needle aspiration (TBNA) is a valuable, minimally invasive procedure for diagnosing and staging lung cancer in patients, but it is underutilized by practicing pulmonologists. To assess the approach to TBNA of current pulmonary Fellows, we recorded their computerized interactive responses during the 1995 American College of Chest Physicians Fellows' Conference. Among 109 Fellows attending, only 10% reported that they routinely (> or = 85% of cases) performed TBNA to diagnose or stage malignant disease, and 40% noted that they rarely (< or = 5% of cases) performed it. They estimated their diagnostic TBNA yields in patients with mediastinal cancer as follows: > or = 80% by 2% of Fellows; between 25% and 80% by 54% of Fellows; and < 25% by 45% of Fellows. They noted that the main limitations of TBNA at their institutions were suboptimal bronchoscopy technique (30%), technician support (1%), cytopathology support (14%), all of these factors (25%), or the belief that TBNA is not useful (30%). TBNA is currently underutilized and/or underemphasized at bronchoscopy training programs. Major modifications of Fellow experiences are necessary if TBNA is to impact optimally on patient management. PMID- 9228386 TI - Hypertrophic obstructive cardiomyopathy: problems of management. PMID- 9228387 TI - Paravertebral mass in a patient with thalassemia intermedia. PMID- 9228389 TI - Lemierre's syndrome in children: high-resolution CT and color Doppler sonography patterns. AB - Lemierre's syndrome is an anaerobic sepsis occurring after oropharyngeal infection in healthy teenagers and young adults. We report two cases of adolescent girls suffering from Lemierre's syndrome studied with cervical color Doppler ultrasonography (CDUS), cervicothoracic helical CT, and high-resolution CT (HRCT) scanning. In both patients, HRCT allowed a good depiction of multiple cavitated pulmonary nodules of various sizes suggestive of this entity and was able to detect small or peripheral nodules. CDUS helped to pinpoint the extent of thromboses of the internal jugular vein demonstrated by CT. CDUS and HRCT should be performed as early as possible to confirm and treat this life-threatening condition. PMID- 9228388 TI - Fever, sore throat, and pulmonary infiltrates in a 20-year-old man. PMID- 9228390 TI - Pulmonary lymphangioleiomyomatosis and cerebrotendinous xanthomatosis: is there a link? AB - A 41-year-old woman had progressive shortness of breath. Cerebrotendinous xanthomatosis was diagnosed 4 years before. An open-lung biopsy showed the simultaneous presence of cerebrotendinous xanthomatosis and pulmonary lymphangioleiomyomatosis. This is perhaps the first time the coincidental occurrence of these two diseases is described. PMID- 9228392 TI - Pneumothorax as a first manifestation of sarcoidosis. AB - Pneumothorax is a rare manifestation of sarcoidosis, occurring usually late in the course of the disease. We report five cases of pneumothorax as a presenting manifestation of sarcoidosis. In two patients, thoracotomy showed extensive pleural infiltration by noncaseating granulomas. High-resolution CT scans showed cavitated subpleural nodules and subpleural bullae in one case. These findings support that necrosis of subpleural granulomas or rupture of a subpleural bullae, or both, are the mechanisms of pneumothorax in sarcoidosis. Three patients with a lung function impairment were treated with oral corticosteroids. One nontreated patient died due to progression of the disease. PMID- 9228391 TI - Role of CT in the management of pneumothorax in patients with complex cystic lung disease. AB - The diagnosis and treatment of pneumothorax in patients with complex cystic lung disease may be difficult when relying on plain chest radiography alone. We report four cases in which management was greatly facilitated by the use of CT scanning of the chest. PMID- 9228393 TI - Recurrence of allergic bronchopulmonary aspergillosis in the posttransplant lungs of a cystic fibrosis patient. AB - Cystic fibrosis (CF) is an autosomal recessive disease of exocrine origin. Allergic bronchopulmonary aspergillosis (ABPA) is an immunologic disorder caused by colonization of the airways with Aspergillus fumigatus. A fumigatus has been cultured from posttransplant lungs in CF patients. Colonization of posttransplant lung with Aspergillus is a recognized phenomenon. In this case report, however, we present a patient who developed ABPA both before and after lung transplant. This patient meets the criteria for ABPA based on serologic results. ABPA may be a complication in post-CF lung transplant patients and serologic analysis should be considered when eosinophilia and pulmonary infiltrates or decline in lung function occurs. PMID- 9228394 TI - Pulmonary smooth muscle proliferation occurring after lung transplantation. AB - We report the first example of smooth-muscle proliferations occurring in an allograft lung implanted in a recipient who had end-stage emphysema. Smooth muscle proliferations were detected 46 months following transplantation in a 53 year-old woman. The lesions involved the airways and were bronchoscopically undetectable. Posttransplant smooth-muscle tumors have been described in liver transplant patients and are thought to be due to Epstein-Barr virus. Evidence of virus infection was not found in the current case. PMID- 9228395 TI - Massive pleural effusion in diffuse granulomatous disease. AB - Although recurrent, massive pleural effusions frequently are associated with malignancy, they rarely occur in granulomatous diseases of the lung. We report the case of a 50-year-old man with a recurrent, massive pleural effusion and a diffuse granulomatous disease that involved the lung, the lymphatics, and bone marrow. The cause of the effusion and granulomatous disease is not entirely clear; however, the effusion did resolve with antifungal therapy and a brief course of corticosteroids. PMID- 9228396 TI - Passive smoking and the 6-minute walk test in heart failure. PMID- 9228397 TI - Pressure-controlled inverse ratio ventilation. PMID- 9228398 TI - Using corticosteroids to treat tuberculous pleurisy. PMID- 9228399 TI - Questioning chest physiotherapy. PMID- 9228400 TI - Pneumothorax ex vacuo. PMID- 9228401 TI - Acute hyperventilation in the emergency department. PMID- 9228402 TI - Improving percutaneous dilational tracheostomy. PMID- 9228403 TI - Fiberoptic bronchoscopy after recent acute myocardial infarction: stress for the heart? PMID- 9228404 TI - A modified outer cannula can help thoracentesis after pleural biopsy. PMID- 9228405 TI - Oxygen desaturation after treatment with inhaled nitric oxide for obstructive shock due to massive pulmonary embolism. PMID- 9228406 TI - Warn asthmatics of scuba diving risks. PMID- 9228407 TI - Wisdom in video-assisted cardiac surgery: what can or should be done? PMID- 9228408 TI - Flesinoxan: a prosexual drug for male rats. AB - Two tests were carried out to compare the stimulatory (i.e., prosexual) effects of the 5-HT1A receptor agonists flesinoxan and 8-hydroxy-2-(di-n propylamino)tetralin (8-OH-DPAT) on sexual behavior in male Wistar rats. Two groups of rats were used: normal males and males with impaired masculine sexual behavior due to neonatal treatment with the aromatase inhibitor 1,4,6 androstatriene-3,17-dione (ATD). In Experiment 1, flesinoxan (0.3 and 1.0 mg/kg) stimulated ejaculation frequency and number of animals displaying this behavior, both in controls and ATD males. With 0.3 mg/kg flesinoxan ATD males did not differ from controls in ejaculation frequencies. There was a concomitant decrease in latency to first ejaculation. No 'premature' ejaculations (i.e., at first or second intromission) were observed. In Experiment 2, the effects of 0.4 mg/kg 8 OH-DPAT, 0.3, 1.0 and 3.0 mg/kg flesinoxan and saline were tested in two ejaculation series. 'Premature' ejaculations only occurred during first ejaculation series with 8-OH-DPAT in 8 of 10 controls and in 6 of 9 ATD males; it did not occur during flesinoxan treatment nor in the second ejaculation series with 8-OH-DPAT treatment. Thus, flesinoxan stimulates sexual behavior in control rats and in rats with impaired sexual behavior. Unlike 8-OH-DPAT flesinoxan does not render them into 'premature' ejaculators. Therefore, flesinoxan could be considered a prosexual drug for male rats. PMID- 9228409 TI - Dizocilpine prevents the enhanced locomotor response to quinpirole induced by repeated electroconvulsive shock. AB - Repeated administration of electroconvulsive shock, as expected, potentiated the locomotor stimulant response to quinpirole (0.3 mg/kg s.c.), a dopamine D2-like receptor agonist. Chronic, but not acute, treatment with the NMDA receptor non competitive antagonist dizocilpine (0.3 mg/kg i.p.) prevented electroconvulsive shock-induced potentiation of quinpirole locomotor response. These results suggest that NMDA receptor activation is necessary for the development of supersensitivity to dopamine receptor agonists produced by repeated electroconvulsive shock. The relevance of this observation in regard to the mechanism of electroconvulsive shock therapeutic effect is discussed. PMID- 9228410 TI - Blockade of clomipramine and amitriptyline analgesia by an antisense oligonucleotide to mKv1.1, a mouse Shaker-like K+ channel. AB - The effect of an antisense oligonucleotide to the K+ channel coding mKv1.1 mRNA on antinociception induced by the tricyclic antidepressants, clomipramine (20-35 mg kg(-1) s.c.) and amitriptyline (10-25 mg kg(-1) s.c.), was investigated in the mouse hot-plate test. Antisense oligonucleotide (0.5-1.0-2.0-3.0 nmol per i.c.v. injection) produced a dose-dependent inhibition of clomipramine and amitriptyline antinociception 72 h after the last i.c.v. injection. The sensitivity to both analgesic drugs returned 7 days after antisense oligonucleotide injection, indicating the absence of irreversible damage or toxicity. Treatment with a degenerated oligonucleotide did not modify the clomipramine- and amitriptyline induced antinociception in comparison with that in naive (unpretreated controls), vector and saline i.c.v.-injected mice. A quantitative reverse transcription polymerase chain reaction (RT-PCR) study demonstrated a reduction in mRNA levels only in the antisense oligonucleotide treated group. Antisense oligonucleotide, degenerated oligonucleotide or vector pretreatment, in the range of doses used, did not produce any behavioural impairment as revealed by the mouse rotarod and hole-board tests. The present results indicate that modulation of the mKv1.1 K+ channel plays an important role in the central analgesia induced by the tricyclic antidepressants, clomipramine and amitriptyline. PMID- 9228411 TI - Increased brain NAD prevents neuronal apoptosis in vivo. AB - Apoptosis is a characteristic form of cell death which has been implicated in neurodegeneration. In this study we document the induction of apoptosis and DNA fragmentation in vivo by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), a neurotoxin. MPTP selectively damages dopaminergic neurons in the substantia nigra of the midbrain. It is a potent inducer of oxygen radicals. Nicotinamide, a precursor of NAD, is able to block the apoptosis induced by MPTP. Nicotinamide also quenches some of the radicals formed by xanthine oxidase. Nicotinamide may be of interest in the treatment of neurodegeneration. PMID- 9228412 TI - Cardiac angiotensin II receptors: studies on functional coupling in Sprague Dawley rats and TGR(alphaMHC-hAT1) transgenic rats. AB - The renin-angiotensin system plays an important role in the pathogenesis of cardiac hypertrophy and chronic heart failure as angiotensin II has been shown to induce cardiac hypertrophy and fibrosis. Besides these structural alterations, functional effects on cardiomyocytes have been reported in different mammalian species. Angiotensin II is known to produce a positive inotropic effect in some species, and differences in atrial and ventricular myocardium have been described. So far, the molecular events which govern angiotensin II-mediated changes in cardiac contractility are not completely understood. In order to study the dependency of the angiotensin II-induced positive inotropic effect on receptor density, we examined the effect of angiotensin II on cardiac function in atria, papillary muscles and isolated ventricular cardiomyocytes from adult Sprague-Dawley rats and TGR(alphaMHC-hAT1) transgenic rats, which expressed the human angiotensin AT1 receptor (hAT1) specifically in the heart. In atrial myocardium from adult Sprague-Dawley rats, angiotensin II (30 micromol/l) produced an AT1-mediated positive inotropic effect (38.5% of control), whereas in papillary muscles and isolated ventricular myocytes, no inotropic response was observed. As shown by polymerase chain reaction (PCR) and radioligand binding, the human angiotensin AT1 receptor was exclusively expressed in transgenic animals, which markedly overexpressed the angiotensin AT1 receptor. However, in transgenic rats the positive inotropic effect in atrial preparations was similar to the controls, and neither in papillary muscles nor in isolated cardiomyocytes the increase in receptor density led to an inotropic effect induced by angiotensin II. These data suggest that the existence of functionally uncoupled receptors rather than the low density of receptors at the ventricular site is responsible for the inability of ventricular myocardium to respond to angiotensin II. PMID- 9228413 TI - Protection against cellular damage in the perfused rat heart by lowered pH. AB - In this comparative study, rat hearts were perfused at 37 degrees C with three clearly defined protocols: the Ca2+ paradox, the O2 paradox and with 20 mM caffeine. Each protocol involved an initial priming (Ca2+(o) depletion or anoxia; stage 1) and subsequent full activation (Ca2+(o) repletion or reoxygenation; stage 2) of the damage system of the sarcolemma. Creatine kinase release in stage 2 was completely inhibited (P < 0.001) in all three protocols when pH was reduced to 6.5 throughout the experiments, or only during stage 1, or only during stage 2. The inhibitor of the Na+/H+ antiporter, amiloride (1 mM), completely prevented creatine kinase release in the Ca2+ paradox (P < 0.001) and markedly reduced damage in the caffeine protocol. Amiloride had no significant effect on creatine kinase release in the O2 paradox. The possible role of Na+(i) was studied in the caffeine protocol: ouabain (5 x 10(-6) M) had little effect whereas substitution of choline for Na+ in the perfusion medium reduced creatine kinase release by about 50%. It is suggested that the same damage system is activated in stage 1 in all three protocols and that a key event is the intracellular production of H+ which are exported via Na+(o)/H(i) exchange. Prevention of H+ efflux by lowered pH(o), even during stage 2, protected against creatine kinase release. The possible role of Na+ movements in the genesis of sarcolemma damage is discussed. PMID- 9228415 TI - Histamine immunoreactivity changes in vestibular-lesioned and histaminergic treated cats. AB - Histamine is likely involved in vestibular function recovery since histaminergic medications are effective in vestibular-related syndromes. We investigated the histamine immunoreactivity changes after unilateral vestibular neurectomy and the effects of betahistine (a partial histamine H1 receptor agonist and an histamine H3 receptor antagonist) and thioperamide (a pure histamine H3 receptor antagonist) treatment in cats. Histamine staining was analyzed in the tuberomammillary and vestibular nuclei through immunohistochemical methods and quantification techniques in light microscopy. Unilateral vestibular neurectomy induced a strong bilateral decrease in histamine immunoreactivity in the vestibular nuclei and a smaller reduction in the tuberomammillary nuclei in both acute (1 week) and compensated (3 weeks, 1 year) cats. One-week thioperamide or betahistine treatment led to a near-total lack of staining in these structures in both lesioned and control cats. One-month betahistine treatment had weaker effects in the compensated cats. We conclude that vestibular lesions reduce histamine staining because of an increase in histamine release in the vestibular and tuberomammillary nuclei, promoting vestibular functions recovery, and betahistine could contribute to this process by acting on both the presynaptic histamine H3 and postsynaptic histamine H1 receptors. PMID- 9228414 TI - Specific attenuation of the pressure-induced contraction of rat cerebral artery by herbimycin A. AB - In order to determine whether protein tyrosine kinase mechanisms are involved in pressure-induced contraction, we compared effects of three structurally unrelated tyrosine kinase inhibitors and orthovanadate, a tyrosine phosphatase inhibitor, on the pressure-induced contraction of the posterior cerebral artery isolated from rats. The change in vessel diameter was continuously measured with a width analyzer. Herbimycin A inhibited the pressure-induced contraction, while it only slightly inhibited contractions produced by potassium chloride or 9,11-dideoxy 11alpha,9alpha-epoxymethano prostaglandin F2alpha (U46619). Genistein inhibited not only the pressure-induced contraction but also the U46619-induced one. Tyrphostin 23 significantly attenuated contractions in response to three different stimuli, i.e., pressure, potassium chloride and U46619. Orthovanadate potentiated the pressure-induced contraction. These results suggest that herbimycin A is a specific and potent inhibitor of the pressure-induced contraction and that a protein tyrosine kinase mechanism may play an important role in the genesis of the pressure-induced contraction of the rat cerebral artery. PMID- 9228416 TI - A post-receptor defect of adenylyl cyclase in severely failing myocardium from children with congenital heart disease. AB - The aim of this study was to determine whether a defect at the post-receptor level of adenylyl cyclase may also contribute to the decreased effectiveness of cAMP-increasing agents in severely failing patients with congenital heart disease. The severity of congestive heart failure in 31 patients with congenital heart disease was graded by a scoring system which included a description of historical and clinical variables. Patients were divided into a group with no or mild heart failure (score < or = 6) and a group with severe heart failure (score > 6). beta-Adrenoceptor-stimulated adenylyl cyclase activity was significantly decreased by 65% in patients with severe heart failure in comparison to the group of patients with no or mild heart failure. In addition, receptor-independent adenylyl cyclase stimulation by forskolin was reduced by 52% in patients with score > 6 compared to patients with score < or = 6. This post-receptor defect of adenylyl cyclase was apparently due to a decrease in the activity of catalytic subunit of adenylyl cyclase as adenylyl cyclase stimulation by forskolin in the presence of Mn2+ which uncouples catalytic subunit from the G proteins, G(s) and G(i), was also significantly diminished in the patients with severe heart failure. In contrast, the level of inhibitory G protein alpha-subunits was apparently not different in the two groups. In summary, the data indicate that a defect at the catalytic subunit of adenylyl cyclase apparently contributes to the decreased effectiveness of cAMP-increasing agents in severely failing patients with congenital heart disease. PMID- 9228419 TI - A piece of my mind. Ambivalence. PMID- 9228418 TI - Effects of CP-060S on membrane channels in vascular smooth muscle cells from guinea pig. AB - The newly developed cardioprotective drug, CP-060S, (-)-(S)-2-[3,5-bis(1,1 dimethylethyl)-4-hydroxyphenyl]-3-[3-[N-methyl-N- [2-(3,4-methylenedioxyphenoxy) ethyl] amino] propyl]-1,3-thiazolidin-4-one hydrogen fumarate, is reported to possess a vasodilating action. Our objective was to examine the effects of CP 060S on the membrane channels in mesenteric arterial cells from guinea pigs, using whole-cell patch-clamp techniques. CP-060S inhibited the Ca2+ channel current in a concentration-dependent manner (ED50 = 1.7 microM at a holding potential of -80 mV and a stimulation frequency of 0.1 Hz). The inhibition was potentiated by a more depolarized holding potential and a higher stimulation frequency. These effects of CP-060S resembled those of diltiazem and gallopamil more than to those of nifedipine; the inhibition was more frequency dependent and less holding-potential dependent than with nifedipine. Higher concentrations of CP-060S also inhibited the delayed K+ channel currents (ED50 = 18 microM). The present observations suggest that CP-060S exhibits the profile of a Ca2+ channel antagonist, similar to that of diltiazem and gallopamil. PMID- 9228417 TI - Monitoring of antisense effects of oligonucleotides targeted to the neuropeptide Y Y1 receptor gene. AB - The suppression of neuropeptide Y Y1 receptor gene expression by antisense oligonucleotides targeted to different gene regions was monitored on mRNA and protein level in the human neuroblastoma SK-N-MC cell line. The antisense oligonucleotide targeted to the junction of the first intron and second exon suppressed specifically Y1 receptor subtype number by more than 50%, but only if oligonucleotides were administered by electroporation. Also, the formation of Y1 receptor mRNA as shown by reverse transcription-polymerase chain reaction was markedly blocked in this case. Using the antisense oligonucleotide targeted to the start of translation, no effect, neither on the Y1 receptor number nor on Y1 receptor mRNA, could be observed. This finding suggests that besides sequence specific effects of antisense oligonucleotides gene site-specific effects play a major role in the efficacy of suppression. PMID- 9228420 TI - 'Hard to reach' Hispanics get health news via physician's radio, TV shows. PMID- 9228421 TI - It takes a community...to lower the STD rate. PMID- 9228422 TI - From the Centers for Disease Control and Prevention. Dog-bite-related fatalities- United States, 1995-1996. PMID- 9228423 TI - From the Centers for Disease Control and Prevention. Tornado-associated fatalities--Arkansas, 1997. PMID- 9228424 TI - The journey back. A physician retrains. PMID- 9228425 TI - Dehydration, delirium, and disability in elderly patients. PMID- 9228426 TI - Radiation therapy for age-related macular degeneration. PMID- 9228427 TI - Radiation therapy for age-related macular degeneration. PMID- 9228428 TI - Criteria for screening blood donors: science or politics? PMID- 9228429 TI - Benefits and risks of screening mammography in women with BRCA1 and BRCA2 mutations. PMID- 9228430 TI - Benefits and risks of screening mammography in women with BRCA1 and BRCA2 mutations. PMID- 9228431 TI - Vaccine safety, media reporting, and Miss America. PMID- 9228432 TI - Disclosure of condom breakage to sexual partners. PMID- 9228433 TI - Surveillance for variant strains of HIV: subtype G and group O HIV-1. PMID- 9228434 TI - The safety of newborn early discharge. The Washington State experience. AB - CONTEXT: While early discharge of newborns following routine vaginal delivery has become common practice, its safety has not been firmly established. OBJECTIVE: To assess the risk for rehospitalization following newborn early discharge. DESIGN: Population-based, case-control study. SETTING: Washington State linked birth certificate and hospital discharge abstracts covering 310578 live births from 1991 through 1994. PATIENTS: Case patients were 2029 newborns rehospitalized in the first month of life. Control subjects were 8657 randomly selected newborns not rehospitalized and frequency matched to case patients on year of birth. Cesarean deliveries, multiple births, and births at less than 36 weeks' gestation were not included. MAIN OUTCOME MEASURE: Stratified analyses and logistic regression were performed to assess the risk for rehospitalization within a month of birth after early discharge (<30 hours after birth) compared with later discharge (30-78 hours after birth). RESULTS: Seventeen percent of newborns were discharged early. Newborns discharged early were more likely to be rehospitalized within 7 days (odds ratio [OR], 1.28; 95% confidence interval [CI], 1.11-1.47), 14 days (OR, 1.16; 95% CI, 1.03-1.32), and 28 days (OR, 1.12; 95% CI, 1.00-1.25) of discharge than newborns sent home later. Subgroups at increased risk for rehospitalization following early discharge included newborns born to primigravidas (OR,1.25; 95% CI, 1.07-1.45), mothers younger than 18 years (OR, 1.22; 95% CI, 0.79-1.91), and mothers with premature rupture of membranes (OR, 1.41; 95% CI, 0.85-2.36). Early discharge was also associated with an increased risk of readmission for jaundice, dehydration, and sepsis. CONCLUSION: Newborns discharged home early (<30 hours after birth) are at increased risk for rehospitalization during the first month of life. PMID- 9228435 TI - Hospital readmission with feeding-related problems after early postpartum discharge of normal newborns. AB - CONTEXT: Increasingly short postpartum hospital stays in the United States precipitated a policy debate that culminated in passage of the Newborns' and Mothers' Health Protection Act of 1996. The debate occurred without population based evidence for adverse health effects in newborns who are discharged early. OBJECTIVE: To determine whether early postpartum hospital discharge of normal newborns increases their risk for hospital readmission with feeding-related problems. DESIGN AND SETTING: Nested case-control analysis of 1991 to 1994 Wisconsin birth certificate and hospital discharge data. SUBJECTS: A total of 210 readmitted case patients and 630 control subjects selected from a cohort of 120 290 normal newborns who weighed at least 2500 g, were delivered vaginally of mothers with uncomplicated medical and obstetrical histories, and were discharged from the hospital either early (day of life 1 or 2) or conventionally (day 3). OUTCOME MEASURE: Readmission at age 4 to 28 days with discharge diagnoses indicating a primary feeding problem, secondary dehydration, or inadequate weight gain. RESULTS: Early discharges increased 3-fold (reaching 521/1000 discharges) during the study period, but feeding-related readmissions (1.7/1000) remained stable. Most readmitted newborns (53.8%) were 4 to 7 days old, many (34.3%) had concurrent dehydration and jaundice, and 29% were admitted through emergency departments. Readmitted newborns were significantly (P<.05) more likely to have been breast-fed, firstborn, or preterm or to have mothers who were poorly educated (<12th grade), unmarried, or receiving Medicaid. Readmission was not associated with early discharge (adjusted odds ratio, 1.05; 95% confidence interval, 0.71-1.53). CONCLUSION: Although several neonatal and maternal factors increase the risk that a normal newborn will be rehospitalized with a feeding related problem, early discharge following an uncomplicated postpartum hospital stay appears to have little or no independent effect on this risk. PMID- 9228436 TI - Long-term risk of tuberculosis among foreign-born persons in the United States. AB - CONTEXT: Cases of tuberculosis (TB) in the United States have declined for 4 consecutive years, but cases among foreign-born persons account for an increasing percentage. OBJECTIVE: To describe the risk of tuberculosis among foreign-born persons with respect to their length of residence in the United States. DESIGN: Cross-sectional analysis of national surveillance data. SETTING: The United States. PATIENTS: All verified TB cases reported to the Centers for Disease Control and Prevention between 1986 and 1994. MAIN OUTCOME MEASURE: Stratum specific incidence rates of TB by age, place of birth, length of residence, age at arrival in the United States, or combinations of these variables. RESULTS: Several groups of persons from countries with a high prevalence of TB had incidence rates higher than 20 per 100,000 person-years more than 20 years after arrival. Among long-term residents, those who arrived in the United States after their fifth birthday had incidence rates of TB 2 to 6 times higher than those of similar age who arrived before their fifth birthday. A total of 45% of the TB cases were among persons younger than 35 years and an additional 18% were among persons who arrived in the United States before their 35th birthday. CONCLUSIONS: Imported Mycobacterium tuberculosis infection (active or latent) is responsible for most TB cases among foreign-born persons in the United States. Detection of active cases among recent arrivals is the main priority in these populations, but many cases were in persons who arrived in the United States before the age of 35 years that could potentially have been avoided with preventive therapy. Elimination of TB in the United States may not be feasible using available diagnostic and treatment modalities without increased efforts to address the global burden of this disease. PMID- 9228437 TI - Primary care physicians' satisfaction with quality of care in California capitated medical groups. AB - CONTEXT: Managed care and capitation have placed new responsibilities on primary care physicians, including formally acting as "gatekeepers" for specialty services and tests. Previous studies have not examined whether primary care physicians who provide services to patients under many coverage arrangements feel differently about caring for patients covered under capitation vs those covered through more traditional forms of insurance. An understanding of whether California primary care physicians feel that they deliver a different level of quality to capitated patients could help signal whether variations in care for patients with different coverage forms are evolving. OBJECTIVE: To evaluate whether primary care physicians in California capitated groups report different satisfaction levels with quality of care for patients in their overall practice than for patients covered by capitated contracts and to examine whether physicians' satisfaction with capitated care quality is influenced by the characteristics of the practice setting. DESIGN: Cross-sectional questionnaire. SETTING: A total of 89 California physician groups with capitated contracts. PARTICIPANTS: A total of 910 primary care physicians (80% response rate). MAIN OUTCOME MEASURE: Satisfaction with 4 aspects of quality of care provided to patients covered by capitated contracts vs patients overall. RESULTS: Physicians reported lower satisfaction with all 4 aspects of care for patients covered by capitated contracts than for patients in their overall practice: 71% were very or somewhat satisfied with relationships with capitated patients (compared with 88% for overall practice), 64% were very or somewhat satisfied with the quality of care they provided to capitated patients (compared with 88% for overall practice), 51% were very or somewhat satisfied with their ability to treat capitated patients according to their own best judgment (compared with 79% for overall practice), and 50% were very or somewhat satisfied with their ability to obtain specialty referrals (compared with 59% for overall practice) (P< or =.001 for all comparisons). Being in a medical group practice (vs an independent practice association) and having a larger percentage of capitated patients were independently associated by multivariate analysis with higher levels of satisfaction with capitated quality of care (P< or =.005). CONCLUSION: These California primary care physicians were less satisfied with the quality of care they deliver to patients covered by capitated contracts than with the quality of care they deliver to patients covered by other payment sources. However, those in medical group practices and with a higher percentage of capitated patients were more satisfied with capitated care. National expansion of capitation should be accompanied by efforts to ensure that the satisfaction of practicing physicians with the care they deliver does not decline. PMID- 9228438 TI - Cholesterol lowering with statin drugs, risk of stroke, and total mortality. An overview of randomized trials. AB - OBJECTIVE: To examine whether cholesterol lowering with 3-hydroxy-3 methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statin drugs) reduces the risks of stroke and total mortality. DATA SOURCES: We conducted a computerized literature search from 1985 through 1995 to identify all published trials testing statin drugs. The Cholesterol and Recurrent Events (CARE) data were added after the report was published in October 1996. Our search was limited to English-language articles and included published overviews containing relevant individual trials. TRIAL SELECTION: Criteria for inclusion of randomized trials in the overview were (1) statin drugs alone used to reduce lipid levels rather than multifactorial interventions including another type of cholesterol-lowering drug and (2) inclusion of data on deaths and/or strokes. DATA EXTRACTION: Data were extracted by 2 researchers, and only minor discrepancies, which were easily resolved by discussion, occurred. Principal investigators of the trials and their funding agencies were also contacted to secure any relevant data not included in the published reports. DATA SYNTHESIS: A total of 16 individual trials including approximately 29 000 subjects treated and followed up an average of 3.3 years were included in the overview. The average reductions in total and low-density lipoprotein cholesterol achieved were large-22% and 30%, respectively. A total of 454 strokes (fatal plus nonfatal) and 1175 deaths occurred. Those assigned to statin drugs experienced significant reductions in risks of stroke of 29% (95% confidence interval [CI], 14%-41%) as well as total mortality of 22% (95% CI, 12% 31%), which was attributable to a significant reduction in cardiovascular disease (CVD) deaths of 28% (95% CI, 16%-37%). There was no evidence of any increased risk in non-CVD mortality (relative risk [RR], 0.93; 95% CI, 0.75-1.14). There was also no significant increase in risk of cancer (RR, 1.03; 95% CI, 0.90-1.17). CONCLUSION: This overview of all published randomized trials of statin drugs demonstrates large reductions in cholesterol and clear evidence of benefit on stroke and total mortality. There was, as expected, a large and significant decrease in CVD mortality, but there was no significant evidence for any increases in either non-CVD deaths or cancer incidence. PMID- 9228439 TI - A 27-year-old woman with migraine headaches. PMID- 9228440 TI - A 61-year-old man with psoriasis, 1 year later. PMID- 9228441 TI - Incremental strategies for providing health insurance for the uninsured. Projected federal costs and number of newly insured. AB - OBJECTIVE: To present several incremental strategies for extending health insurance coverage for segments of an estimated 40.6 million uninsured persons in the United States. Along with these strategies, the federal costs and estimates of the number of newly insured are presented. DESIGN: Using data from the Congressional Budget Office and the federal government, the number of newly insured persons in the United States under options designed to increase coverage among uninsured children, their parents, and workers between jobs are simulated. The federal costs and coverage implications of these options are estimated for federal fiscal years 1998 through 2002. METHODS: Three distinct incremental approaches for covering the uninsured are explored. The first approach would expand coverage through the current Medicaid program. The second approach would provide financial incentives for parents of children eligible for Medicaid to purchase coverage, and the final approach provides time-limited subsidies allowing workers and their families to purchase insurance when they are between jobs. MAIN RESULTS: The federal costs of these approaches range from $2 billion to $3 billion per year (enrollment outreach approach) to $5 billion to $7 billion per year (enrolling parents of Medicaid-eligible children approach). If pursued simultaneously, the incremental strategies under investigation could extend health insurance to more than 7 million uninsured persons in the United States. The cost of these options could be financed through Medicaid savings, restructuring the current disproportionate share payments made through Medicare and Medicaid, increasing excise taxes on tobacco, or all of the above. CONCLUSIONS: The incremental strategies would build on the current US health care delivery system by providing targeted financial assistance to specific populations. By their nature, such reforms could provide a political means for compromise and agreement between Congress and the president. Though the reforms do not, by design, provide a comprehensive solution to the problems facing the uninsured, they would address the severe problems facing many low- and middle income families unable to purchase health insurance today. PMID- 9228442 TI - Early discharge and evidence-based practice. Good science and good judgment. PMID- 9228444 TI - Total quality management and the culture of organization. PMID- 9228443 TI - Preventing opportunistic infections in persons infected with HIV: 1997 guidelines. PMID- 9228445 TI - Repeated psychiatric referrals to Belgian emergency departments: a survival analysis of the time interval between first and second episodes. AB - Repetition of psychiatric emergency department use by a relatively small number of patients constitutes a major problem for clinicians and service providers. This study aimed at the identification of risk factors for repetition by addressing the time interval between the first and second visits to the emergency department. The purpose was to investigate what patient characteristics and referral circumstances determine this interval. Over a two year period, data on all psychiatric emergency referrals to the emergency department of four public hospitals were collected with a standardized form. Data collected during the index referral of all patients were used for estimating the risk for repetition using survival analysis techniques. A large proportion of repeaters revisits the emergency department within a short time interval. Younger, male patients who present themselves spontaneously are more likely to repeat than others. Previous inpatient service use and the presence of a diagnosis of substance abuse disorder or psychotic disorder at the first visit further increases the risk for repetition. Previous service use and, to a lesser degree, demographic and clinical characteristics of psychiatric patients are useful in the prediction of variations in time between first and second referrals to the emergency department. PMID- 9228446 TI - What do accident and emergency medical staff think of practice guidelines? AB - The objective of this paper was to determine the views of accident and emergency consultants and trainees towards practice guidelines and their experiences using guidelines. A postal questionnaire survey of consultants, senior registrars and registrars in accident and emergency medicine was carried out in Yorkshire. The results of this survey show that the potential benefits of practice guidelines are appreciated, and that evidence-based and 'user-friendly' guidelines are wanted. It is concluded that unless rigorously developed and clear and easy to use, guidelines are unlikely to be implemented in accident and emergency departments in the UK. PMID- 9228447 TI - Patients discharged from emergency care after acute myocardial infarction was ruled out: early follow-up in relation to gender. AB - The aim of this research was to describe men and women who were discharged from the emergency department after having an initial suspicion of acute myocardial infarction ruled out in terms of patient characteristics, symptom reevaluation, electrocardiogram and exercise stress test. Consecutive patients below the age of 65 years who came to the emergency department of Sahlgrenska Hospital with acute chest pain or other symptoms raising suspicion of acute myocardial infarction for whom the suspicion was ruled out either directly in the emergency department or less than 1 day after hospital admission were included in the study. Four hundred and eighty-four patients participated, of whom 295 (61%) were men. Men had a higher prevalence of ischaemic heart disease. The cause of pain was judged similarly at reevaluation compared with in the emergency department in 53% of the cases. Only in 4.6% of the cases were the symptoms judged to be caused by myocardial ischaemia on both occasions. At the initial visit 36.0% of the patients were judged to have uncertain cause of the symptoms. This proportion was lowered to 26.4% at reevaluation. The exercise electrocardiogram at reevaluation revealed clinical and electrocardiographic signs indicating definite myocardial ischaemia in 2.6% of the cases. Early follow-up of patients discharged from the emergency department after acute myocardial infarction was ruled out revealed that a low proportion showed signs of myocardial ischaemia. In about half of the cases the judgement differed from that being made in the emergency department. PMID- 9228448 TI - Outcome of cardiopulmonary resuscitation in a Portuguese university hospital. AB - Portugal has many weak links in the so-called 'chain of survival' both in the pre hospital and in-hospital setting. Apart from evaluating the performance of a newly implemented in-hospital cardiac arrest system, we assessed the correlation between different clinical variables and outcome after cardiopulmonary resuscitation (CPR). All resuscitation attempts during 1995 were registered using the form recommended by the European Resuscitation Council. One hundred and twenty-five patients either collapsing out-of-hospital (42; 33.6%) or having in hospital cardiac arrest (83; 66.4%) were included in the study. The number of patients who experienced recovery of spontaneous circulation was 10 (23.8%) in the out-of-hospital group and 35 (42.2%) in the in-hospital one. Of all out-of hospital patients, 11.9% were discharged from hospital compared with 22.8% of the in-hospital group. Asystole was the initial rhythm in a significant number of arrests. The average call-response interval of the in-hospital cardiac arrest team was under 3 min. The results from this series concur with other reported series. Although good standards of care were achieved, we are aware that this was only an isolated step in the implementation of the 'chain of survival' in our country. The authors conclude that there is an urgent need for a nationwide programme that improves the standards of care for these patients. PMID- 9228449 TI - Spiral computed tomography with three-dimensional reconstructions for severe blunt abdominal traumas: a useful complementary tool? AB - Spiral computed tomography (CT) has proved to be a valuable tool by providing three-dimensional (3D) images of the studied structures. We hypothesized that a more realistic depiction of lesions by 3D CT could be of interest for surgeons who are treating blunt abdominal traumas and lead to less inappropriate triage. A good working relationship between surgeons and radiologists allowed us to perform a 3D CT examination in six patients. In the first patient, the 3D CT accurately demonstrated spleen fragmentation without devascularized fragment. The second patient had complete devascularization of the spleen upper pole. Conservative treatment was pursued for both patients. For the third patient, 3D CT helped us to differentiate peritoneal-perisplenic fluid from subcapsular fluid. The fourth patient had minor spleen injury associated with severe lacerations of the left kidney. 3D CT showed a complete separation of the kidney lower pole. A delayed partial lower nephrectomy was performed. The fifth patient presented a fragmented spleen and transient massive haematuria related to a well-contained laceration of the kidney upper pole that were amenable to nonoperative management. The sixth patient was emergency operated for active bleeding from a fragmented spleen. 3D CT performed 2 months after spleen repair allowed the assessment of the amount of devascularized tissue, as well as the status of the upper abdomen arteries. For haemodynamically stable patients, 3D CT could be a helpful addition to conventional axial CT for quantifying blunt abdominal traumas, for making the choice between nonoperative and operative treatment, but also between emergency and delayed surgical strategy. PMID- 9228450 TI - Videotaping in the admitting area: a most useful tool for quality improvement of the trauma care. AB - The treatment of trauma patients in the admitting area is performed under stress and requires team work. The goals of this research were to develop and analyse the implementation process of videotaping trauma care. The Rambam Medical Center is a 900-bed referral teaching hospital. It serves a population of more than 1.5 million in northern Israel. The trauma unit has focused on various activities to increase the quality of trauma care over the past few years. We installed a video camera and taped the treatment as part of a programme for the quality improvement of trauma care. Reviewing the tapes was carried out by the trauma team under guidance in order to identify deviation from treatment protocols, errors in techniques, improper usage of time, equipment failure and problems in team work. After 3 years' experience, we found that videotaping is an accurate and inexpensive way of achieving quality control in the admitting area. It now serves as a regular method in Israeli trauma centres and we encourage others to try this method. PMID- 9228451 TI - Evaluation of trauma care in a developing country highlighted by a major aircraft accident. AB - The aim of this study was to evaluate the day-to-day trauma care in a developing country highlighted by a major accident. In this accident, early management was not carried out according to triage principles. Scene mortality and in-hospital mortality were 72% (n = 55) and 14% (n = 3), respectively. Overall mortality rate was 76%. Five survivors were minor wounded. Three laparotomies, one thoracotomy and three tube thoracostomies were performed in the acute phase. Skeletal injuries, mainly rib fractures (43.3%) and haemothorax (10.8%), were the most frequent pathologies seen. One liver laceration, one splenic rupture, one intraabdominal bleeding due to rupture of mesenteric vessels, two major cranial traumas and an abruptio placenta were the other pathologies. The missed injury rate in this accident was 16% (n = 6). It is concluded that the missed injuries in this incident reflect the inadequacy of trauma care in the rural area of the developing country. PMID- 9228452 TI - Management of acute confusion in the elderly. AB - Acute confusion in the elderly as a presenting symptom in an accident and emergency department requires just the same energy devoted to diagnosis as does, say for example, acute coma, epilepsy or haematemesis. Doctors in accident and emergency departments are reminded not to succumb to the pitfalls of assuming that acute confusion is merely part of a progressive dementia in an elderly person and therefore incapable of treatment. In this paper a number of clinical examples are given where treatment of the cause of the acute confusion has led to restoration of the patient's independent existence. In passing, the abbreviated mental test score is commended to accident and emergency doctors as being just as useful in a different context as the worldwide acceptance has been of the Glasgow Coma Score. Currently the standard abbreviated mental test score seems confined to the United Kingdom as part of the generally accepted practice. PMID- 9228453 TI - Tools for evaluating disasters: preliminary results of some hundreds of disasters. AB - Epidemiologic research of disasters is hampered by a lack of uniformity and standardization in describing these events. By applying a classification and scoring system, which recently became available, an analysis could be performed of 416 disasters from the past 40 years. Only 79 references were useful in obtaining reliable figures for a scoring on the Disaster Severity Scale (DSS). The various disaster types show a relationship between the DSS-scoring on the one hand, and the severity factor (S) and the number of dead and wounded (n) on the other. It is concluded that the classification and scoring system used could serve as a tool for evaluating the majority of disasters. A small improvement of this system is recommended. PMID- 9228454 TI - Fatal Streptococcus viridans descending mediastinitis: case report and review of the literature. AB - Mediastinitis is a life-threatening complication of cardiac, neck and oesophageal surgery. It has also been reported following upper digestive and respiratory procedures and as a consequence of oesophageal perforation following the ingestion of foreign bodies. Much more infrequently, mediastinitis can occur in association with oropharyngeal or cervical infections. We describe the case of a patient with fatal mediastinitis and septic shock. The onset of mediastinitis was preceded by a 2-day course of sore throat and other flu-like symptoms. PMID- 9228455 TI - Isolation and partial characterization of LH, FSH and TSH from canine pituitary gland. AB - A new preparative procedure without using ion-exchanger is described for the efficient purification of canine LH (cLH), FSH (cFSH) and TSH (cTSH) from the pituitary gland. The hormones were extracted from the pituitary homogenate with an ammonium sulfate solution, and were separated by Concanavalin (Con) A affinity , hydrophobic interaction-, then immobilized metal ion affinity chromatography. In the immobilized metal ion affinity chromatography, we used copper (Cu2+) as chelated metal ion with ammonium ion gradient and pH gradient in phosphate buffer to attain separation of the hormones. High purity of cLH, cFSH and cTSH was indicated as single bands in SDS-PAGE, with apparent molecular masses of 34, 36 and 37 kDA, respectively. The purified hormones showed two bands corresponding to alpha (20 kDa) and beta subunits (cLH beta: 16 kDa, cFSH beta: 22 kDa, cTSH beta: 16 kDa) under reducing condition in SDS-PAGE. The purified hormones were prepared in good recovery (LH: 53%, FSH: 34%, TSH: 36%) with high biological activity or binding activity to the receptor. Cross-contamination of the purified hormone was less than 0.5%. Examination of the hormone fraction with isoelectric focusing showed that major peaks of isoelectric isoforms were maintained throughout the purification steps of cLH and cFSH, while a few peaks were lost in Con A affinity chromatography in cTSH purification. It was concluded that the present method could prepare highly purified cLH, cFSH and cTSH which retained isoforms of the hormones and biological activity or binding affinity to the receptor. PMID- 9228456 TI - The pressor response induced by repeated injections of neostigmine into the central nervous system is desensitized in adrenalectomized, but not in intact rats. AB - To investigate whether or not pressor responses to repeated stimulation of central cholinoceptive neurons are desensitized, mean arterial blood pressure (MAP) and heart rate (HR) were measured following repeated injections of neostigmine, an acetylcholinesterase inhibitor, into the third cerebral ventricle in conscious, unrestrained intact or adrenalectomized (ADX) rats. Neostigmine (5 x 10(-9) or 5 x 10(-8) mol) in 1 milliliter saline increased MAP dose-dependently and increased HR in intact rats. The peak values in MAP and HR after three repeated injections at 4 hour intervals did not wane. Neostigmine (5 x 10(-8) mol) also increased MAP in ADX rats, and the peak values after the first injection were higher in the ADX rats than in the intact rats. The pressor responses to the second and third injection, however, were less than to the first injection in the ADX rats. HR responses to the repeated injections in the ADX rats were identical to those in the intact rats. These findings suggest that the adrenal gland plays a role in antagonizing the development of desensitization in the neostigmine-induced pressor response. PMID- 9228457 TI - Two cases of acromegaly in a family. AB - We report two cases of acromegaly due to pituitary adenoma without any other endocrinopathy in a family. The patients had high plasma GH and were improved by transsphenoidal adenomectomy. Acromegaly is usually a clinical syndrome of sporadic nonfamilial occurrence. The familial occurrence of acromegaly not associated with multiple endocrine neoplasia is very rare. Our patients are unlikely to be associated with the multiple endocrine neoplasia type 1 syndrome. Here we describe two patients with acromegaly, a father and his daughter, and review familial cases reported. PMID- 9228458 TI - Clinical courses and thyroid conditions in three infants born to a mother with thyroid stimulating-blocking antibodies. AB - The clinical courses including thyroid conditions of three infants born to a mother with primary hypothyroidism due to Hashimoto's thyroiditis were studied. The mother was positive for both TSH-binding inhibitor immunoglobulins (TBII) and thyroid stimulating-blocking antibodies (TSBAb) in her serum. The first infant died because of septic shock due to fistula formation between the large intestine and the bladder. Serum thyroid hormone levels during the first pregnancy were extremely low because of incomplete replacement therapy with levothyroxine. The second infant had almost normal thyroid function, so that the replacement therapy was not necessary. The third infant had transient and overt primary hypothyroidism. The replacement therapy was carried out for six months after birth. TSBAb activities in this mother were high in the third pregnancy. In general, these activities gradually increases with the clinical course in TSBAb positive Hashimoto's patients. From these findings, it was suspected that the thyroid conditions in the second and the third infants reflected the natural course of TSBAb activities in this mother. PMID- 9228459 TI - Effects of dietary fructose or glucose on triglyceride production and lipogenic enzyme activities in the liver of Wistar fatty rats, an animal model of NIDDM. AB - Effects of dietary carbohydrates on triglyceride production and hepatic lipogenic enzyme activities were examined in Wistar fatty rats, an animal model of noninsulin dependent diabetes mellitus, fed fructose or glucose and were compared with those of Wistar lean rats. Carbohydrates were supplied in 10% drinking solutions for 21 days. As compared with lean rats, Wistar fatty rats were characterized by hyperglycemia, hyperinsulinemia and hypertriglyceridemia, the last of which was associated with an increased hepatic activity of fatty acid synthetase and an increased rate of triglyceride secretion from the liver to the circulation. Feeding fructose to genetically obese diabetic rats produced a threefold increase in the hepatic activity of fatty acid synthetase, a twofold increase in NADPH-generating enzymes (malic enzyme and glucose-6-phosphate dehydrogenase) and a 56% increase in the rate of triglyceride secretion, with a resultant 86% increase in plasma triglyceride concentrations. Feeding glucose produced a similar increase in the activity of NADPH-generating enzymes and triglyceride production in the fatty liver but it differed in producing no change in plasma triglyceride concentrations or hepatic fatty acid synthetase activity. Neither dietary fructose nor glucose changed glycemia or insulinemia. These results show that in genetically obese, diabetic rats feeding fructose and glucose is associated with an increase in hepatic lipogenic enzyme activities and triglyceride production, and suggest that fructose stimulates triglyceride production but impairs triglyceride removal, whereas glucose stimulates both of them. PMID- 9228460 TI - Cloning and characterization of the 4.2 kb region of the rat thyrotropin receptor promoter. AB - We previously identified an approximately 200 bp "minimal promoter" of the rat TSH receptor (TSHR) gene which is essential for the promoter activity. In the present study, we have cloned and characterized an upstream region of the TSHR promoter to disclose additional functional element(s). We screened a rat genomic library and obtained a DNA fragment which contained a 4.2 kb 5'-flanking region. This fragment was 2.5 kb longer than that we previously studied (1.7 kb). To assess the promoter activity, chimeric plasmids containing the 4.2 kb promoter and its 5'-deletions ligated to a chloramphenicol acetyltransferase gene were transfected into thyroid and non-thyroid cells. These plasmids expressed significant promoter activity in FRTL-5 and FRT thyroid cells, but not in BRL liver cells. The strongest promoter activity was expressed by the -199 bp promoter, and the longer promoter expressed rather decreased activity. Co expression of thyroid transcription factor-1 (TTF-1) increased the activity of the promoter region from -3187 to -199 bp, which encompassed one or two TTF-1 binding sites we previously identified, but not the -4206 bp promoter. In addition, FRTL-5 stable transfectants each having a chimeric construct were cultured in the presence or absence of TSH. All transfectants expressed higher promoter activity in the absence of TSH than in the presence of TSH, in particular, the -3187 bp plasmid expressed significantly higher activity by comparison to the -2617 and -4206 bp constructs. This result indicates that the region between -3187 and -2617 bp may contribute to TSH/cAMP-induced suppression and also suggests that the region between -4206 and -3187 bp involves the element(s) for constitutive suppression of the promoter activity. These results not only suggest that the 4.2 kb upstream region of the TSHR gene possibly contains some elements for the regulation of the gene expression, but also emphasize the importance of the minimal promoter region which we previously identified for the efficient expression of the gene. PMID- 9228461 TI - Acute effect of prolactin on bone 45Ca accumulation in rats. AB - In an attempt to evaluate the acute effect of PRL on bone turnover, experiments were performed in sexually mature female Wistar rats which were given an intraperitoneal (i.p.) administration of 1.25 mM CaCl2 solution containing 6 microCi 45Ca. Dose response study showed a two fold higher rate of 45Ca uptake in 30 min by control trabecular bones (sternum and vertebrae 5-6) compared with control compact bones (femur and tibia). Femur exhibited a dose dependent increase in 45Ca uptake in response to pharmacological doses of 0.01 and 0.02 mg PRL/100 g body weight but was less responsive to 0.04 mg PRL/100 g body weight. Tibia only responded with increased 45Ca uptake to 0.02 and 0.04 mg PRL/100 g body weight while trabecular bones were unresponsive. By varying the intervals between administration of 45Ca (0 min) and PRL (0, 30 or 60 min) and bone harvesting (30, 60 or 90 min), it was found that 0.01 mg PRL/100 g body weight had a biphasic action on 45Ca movement. It initially enhanced 45Ca influx into femur by 30 min followed by an accelerated 45Ca efflux from femur back to the circulation. It could be concluded that PRL acutely stimulated calcium turnover in compact bone. PMID- 9228462 TI - Detection of novel molecules recognized by anti-placental lactogen antibody in rat amniotic fluid. AB - Amniotic fluid contains various bioactive substances including the placental PRL family. In the present study, it was elucidated that rat amniotic fluid contained immunoreactive proteins which had different molecular sizes and pI values from the authentic placental lactogens (PLs) in the rat, recognized by antipeptide antibody to the N-terminal peptide of rat PL-I. Immunoreactive PLs residing in the amniotic fluid were characterized further by two-dimensional sodium dodecyl sulfate gel electrophoresis (2DE), immunoblotting and anion-exchange chromatography. Amniotic fluid collected from rats on day 12 of pregnancy contained two PL-like molecules, tentatively called A1 (MW 75 kDa, pI 4.6) and A2 (MW 99-102 kDa, pI 5.3-5.4). A1 and A2 are specific to the amniotic fluid, because no such molecules were found in the serum or placental extracts. Immunoblot analysis of amniotic fluid revealed that A1 levels increased, whereas those of A2 decreased to an undetectable level up to day 16 of pregnancy. When the A1 concentrations from days 12 to 20 were monitored intensively, they increased from day 12 to 14, were maintained until day 18, and then decreased dramatically by day 20. The expression pattern for A1 was therefore completely different from those of authentic placental PRL family members found in serum and placental tissue, indicating that the A1 is distinct from them. Partial purification by anion-exchange chromatography and 2DE revealed that A1 consisted of 5 isoforms. PMID- 9228463 TI - Long-term follow-up of a girl with the neonatal form of Bartter's syndrome. AB - We followed up a girl with the neonatal form of Bartter's syndrome for sixteen years and determined the sensitivity to angiotensin II before and during the indomethacin treatment. A 4-month-old girl was admitted to our hospital, because of severe hypokalemia and growth retardation. Initially we treated her with spironolactone and potassium supplements. This treatment increased plasma potassium levels and her growth. At the age of one year she was diagnosed as having Bartter's syndrome. Since then she has been treated with indomethacin at an initial dose of 3 mg/kg/day combined with spironolactone and potassium. After the start of the indomethacin treatment, her growth increased dramatically, and her final height was normal adult height. Her puberty developed normally and menarche occurred at the age of 12 years. Levels of serum sodium, chloride, plasma aldosterone and urinary prostaglandin E2 were also normalized. Levels of angiotensin I and II were improved but not within the normal range, but plasma potassium levels slightly decreased after plasma aldosterone levels were normalized and did not change during the treatment period. Plasma renin activity remained high until about the age of 8 years, after which it decreased to almost within the normal range. At 5 months after the start of indomethacin (3 mg/kg/day), her vascular sensitivity to angiotensin II had been improved, and after 2 years and 5 months, her vascular sensitivity was further improved. At this time renin activity had decreased after angiotensin II infusion, but plasma aldosterone did not change. At the age of 16 years (dose of indomethacin: 0.5 mg/kg/day), plasma aldosterone increased after angiotensin II infusion. These data suggest that indomethacin and spironolactone are effective treatments for the neonatal form of Bartter's syndrome, especially during childhood. PMID- 9228464 TI - Galanin exerts dual action on inositol-specific phospholipase C activity in isolated pancreatic islets. AB - The intracellular mechanism whereby the neuropeptide galanin inhibits insulin secretion is not establish, since the peptide affects several signal pathways, including intracellular messengers such as calcium and cyclic AMP. In this study, we have assessed the effect of galanin on the inositol-specific phospholipase C (iPLC) activity in isolated rat pancreatic islets. The iPLC activity was measured as the generation of inositol 1,4,5-trisphosphate and its metabolite inositol 1,3,4-trisphosphate from the hydrolysis of polyphosphoinositides. Inositol phosphates were measured by anion-exchange fast protein liquid chromatography (FPLC) analysis of extracts from islets prelabelled with myo-3H-inositol. Galanin (1 to 100 nM) significantly increased the glucose-induced (12 mM) accumulation of inositol 1,4,5-trisphosphate after 2 min, but this stimulation of iPLC activity was followed by a significant suppression after 15 min. In the absence of extracellular calcium, both the stimulatory and inhibitory effects of galanin on the iPLC activity vanished. We therefore conclude that galanin initially stimulates iPLC in a calcium-dependent manner, followed by a secondary inhibitory effect. The secondary inhibition of iPLC activity might contribute to the insulinostatic action of the neuropeptide. PMID- 9228465 TI - A study on the biological significance of midregion and intact parathyroid hormone in hemodialysis patients. AB - In this study, we investigated the relationship between the concentrations of intact parathyroid hormone (i-PTH) and midregion PTH (m-PTH) measured by an immunoradiometric assay and a radioimmunoassay, respectively, versus various demographic and biochemical parameters, bone mineral density (BMD) of the lumbar spine (LS) and radius, and the radiographic findings of osteosclerosis and aortic calcification in hemodialysis (HD) patients. m-PTH correlated positively and more significantly with serum calcium (Ca), serum phosphorus (P), Ca-P solubility products (Ca x P) and LS-BMD than i-PTH did (P = 0.024 vs. 0.531, 0.001 vs. 0.061, 0.0001 vs. 0.125, and 0.017 vs. 0.284, respectively). A positive correlation between the percent changes in serum P over the 1-month measurement period and those in m-PTH rather than in i-PTH was also observed (P = 0.021 vs. 0.869). These data indicate than m-PTH is distinct from i-PTH in its positive correlation with serum Ca, serum P, Ca [symbol: see text] P and LS-BMD in HD patients. Since m-PTH is known to consist mostly of the midregion and carboxyl terminal fragments of PTH in HD patients, the present study suggests that these PTH fragments may be biologically significant in the patients in vivo. PMID- 9228466 TI - Effect of 1 alpha-hydroxyvitamin D3 on loss of bone mineral density immediately after artificial menopause. AB - To evaluate the effects of 1 alpha-hydroxyvitamin D3 (1 alpha(OH)D3), a series of clinical trials, preventive and therapeutic, were performed in an open label manner in women immediately after oophorectomy. The series included a total of 121 oophorectomized subjects, whose lumbar bone mineral density (L2-4BMD) was followed by the use of dual energy X-ray absorptiometry. (1) Preventive trial: 61 women who had undergone premenopausal bilateral oophorectomy, were divided into 3 groups (Group C: control; Group L: 0.25 micrograms 1 alpha (OH)D3/day; Group H: 0.50-0.75 micrograms 1 alpha (OH)D3/day). The changes in BMD and chemical indices were followed up for one year. (2) Therapeutic trial: the trial included 60 premenopausally oophorectomized subjects having L2-4BMD lower than the normal control level minus 1SD which has been reported in age-matched normal Japanese women. These subjects were divided into 3 groups and treated in the same way as in the preventive trial. In the preventive trial, L2-4BMD decreased by 8.2%, 6.5% and 4.5% in groups C, L and H, respectively, at 12 months of treatment, whereas in the therapeutic trial, L2-4BMD decreased by 3.6%, 3.2% and 0.8% in the groups C, L and H, respectively, at 12 months of treatment. In conclusion, 1 alpha(OH)D3 was found to be effective both to prevent the bone loss subsequent to bilateral oophorectomy and improve low bone mass after oophorectomy. PMID- 9228467 TI - Octreotide and bromocriptine suppress thyroid hormone levels and thyroid nodule in an acromegalic patient with nontoxic autonomous goiter. AB - An acromegalic patient with nontoxic autonomous goiter was sequentially treated with octreotide and bromocriptine. Before therapy, serum GH, PRL and insulin-like growth factor-I (IGF-I) levels were increased. Free T3 and free T4 were within the normal range with suppressed TSH levels, whereas 123Iodine-uptake of thyroid was 5.6% after 24 h. During treatment with octreotide and bromocriptine, serum GH, PRL, and IGF-I became normal and free T3 and free T4 were slightly but significantly decreased, but TSH levels remained very low. After thyroidectomy, thyroglobulin, free T3 and free T4 were further decreased, and the TSH levels were recovered to normal. These findings suggested that octreotide and bromocriptine inhibit the release of thyroid hormones from the autonomous thyroid gland directly or indirectly through the decline in IGF-I. PMID- 9228468 TI - Interaction of osmotic and nonosmotic stimuli in regulation of vasopressin secretion in hypoosmolar state of man. AB - Vasopressin (AVP) secretion is principally under osmotic regulation, which is altered by nonosmotic stimuli. It is known that the manner of osmotic regulation of AVP secretion in hypoosmolar state of man consists of four types. The types have (A) random changes in plasma AVP without relation of plasma osmolality; (B) plasma AVP secretion correlated closely to plasma osmolality with a low osmotic threshold for AVP release; (C) nonsuppressible AVP secretion with normal osmotic release of AVP; (D) no abnormalities in AVP secretion. In this study, we found an entirely different type of AVP secretion from the above types in six patients with hyponatremia resulting from various causes during infusion of 5% hypertonic saline. To clarify the mechanism underlying the AVP secretion, we analyzed the interaction between osmotic and nonosmotic stimuli of AVP secretion in these patients. Despite hyponatremia, plasma AVP levels in all patients were not suppressed, which was attributed at least in part to the presence of nonosmotic stimuli for AVP release. These stimuli include nausea, hypotension, blood volume contraction, glucocorticoid deficiency or their combinations. Hypertonic saline infusion increased both serum sodium concentrations and plasma osmolality, although to subnormal levels, and concomitantly, alleviated some of the nonosmotic stimuli for AVP release formerly present in these patients. However, plasma AVP concentrations decreased rapidly during the infusion and reached the nadir in all patients. This phenomenon may be due to alleviation of nonosmotic stimuli for AVP release. Thus, the findings indicate that the potentiating effect of nonosmotic stimuli for AVP secretion may modify the osmotic regulation of AVP secretion in hypoosmolar state, resulting in the type of AVP secretion in this study. PMID- 9228469 TI - Plasma chromogranin A in pheochromocytoma, primary hyperparathyroidism and pituitary adenoma in comparison with catecholamine, parathyroid hormone and pituitary hormones. AB - Plasma levels of chromogranin A (CgA) were measured by ELISA in 22 patients with pheochromocytoma (18 non-metastatic, 3 metastatic, and 1 mixed neuroendocrine neural tumor), 9 patients with primary hyperparathyroidism, and 9 patients with pituitary adenoma. The plasma levels of CgA were compared with norepinephrine, epinephrine, parathyroid hormone and pituitary hormones, i.e., growth hormone and prolactin. In pheochromocytoma, CgA in preoperative plasma of the patients without metastasis was 228 +/- 38 U/L (mean +/- SEM) and significantly higher than healthy controls (30 +/- 11 U/L, n = 40). Plasma CgA was decreased after removal of the tumors (28 +/- 6.0 U/L), except in three patients with metastatic pheochromocytoma and a mixed neuroendocrine neural tumor. The concentration of CgA in the patients with non-metastatic pheochromocytoma was significantly correlated with that of plasma norepinephrine (P < 0.005, r = 0.68) and urinary norepinephrine (P < 0.05, r = 0.65), but not with that of epinephrine. There was an exceptional case in which CgA was extremely high, but the CA level was normal. This tumor was a highly malignant pheochromocytoma with extensive metastases composed of small tumor cells which were occasionally positive for tyrosine hydroxylase immunohistochemically. These cells were considered to be poorly differentiated tumor cells and synthesized a very small amount of norepinephrine. Plasma levels of the patients with primary hyperparathyroidism and the patients with pituitary adenoma were 44 +/- 4 U/L and 48 +/- 8 U/L, respectively. Only one patient with a growth hormone-producing pituitary adenoma had a high level of CgA. Plasma CgA is a useful tumor marker for pheochromocytoma, even for malignant pheochromocytoma without elevated CA level, but not for hyperparathyroidism, or pituitary adenoma. PMID- 9228470 TI - A case of nonfunctioning pituitary adenoma resembling so-called silent corticotroph adenoma. AB - It has been indicated that some subsets of clinically nonfunctioning pituitary adenoma exhibit immunohistochemical and ultrastructural features of somatotroph or corticotroph cells, which are described as silent somatotroph and corticotroph adenomas, respectively. We here describe a 70-year-old woman with nonfunctioning pituitary adenoma presenting with visual disturbance. None of the pituitary hormones show abnormal finding. Magnetic resonance imaging study revealed a pituitary tumor which developed inferiorly into the sphenoidal sinus. Tumor resection was performed by a transsphenoidal approach, and resulted in a partial resection of the tumor. The specimen indicated chromophobic adenoma. Immunohistochemical examination revealed a characteristic distribution pattern of cytokeratin staining for ACTH cell adenoma, although immunostaining for ACTH was only weakly positive. After the operation, only ACTH secretion was impaired, requiring replacement therapy with glucocorticoid. Bromocriptine was administered in order to prevent the recurrence of the tumor. It remains to be elucidated whether the present case could be classified as silent corticotroph adenoma, which was originally indicated to have aggressive characteristics, i.e., progressive visual defect, high incidence of infarction and frequent recurrence. PMID- 9228471 TI - Potentiating effects of calcium chelators on basal and stimulated aldosterone production by bovine adrenal glomerulosa cells in vitro. AB - We previously reported the potentiation of basal aldosterone production by the addition of the calcium chelator ethyleneglycol-bis (beta-aminoethyl ether) N,N,N',N'-tetraacetic acid (EGTA) to an extracellular solution of bovine adrenal glomerulosa cells in vitro. To assess whether the addition of the calcium chelators ethylenediamine-N,N,N',N'-tetraacetic acid (EDTA) and EGTA can potentiate basal and stimulated aldosterone production, we compared the effect of EDTA with that of EGTA on basal and the agonist (potassium; 8 mM, angiotensin II; 10 nM, ACTH; 10 nM)-stimulated aldosterone production by the cells in vitro. These two chelators lowered the extracellular ionized calcium ([Ca2+]o) concentration in a similar manner. The levels of basal and the agonist-stimulated aldosterone production in the presence of EDTA (1 mM) and EGTA (1 mM) were significantly (P < 0.01 or less) increased when compared with those in the absence of EDTA and EGTA, respectively. These results show that the addition of EDTA and EGTA to an extracellular solution potentiates basal and the agonist stimulated aldosterone production in vitro. Although an increase in basal aldosterone production in the presence of EDTA (1 mM) and EGTA (1 mM) was completely inhibited by the voltage-dependent calcium channel antagonist nifedipine (1 microM) or the calmodulin antagonist pimozide (1 microM), the potentiation of the agonist-stimulated aldosterone production does not seem to be induced by CA2+/calmodulin-dependent nor cAMP-dependent systems. These findings suggest that calcium chelators such as EDTA and EGTA may possess activating effect on basal and stimulated aldosterone production in bovine adrenal glomerulosa cells. PMID- 9228472 TI - Multicenter randomized trial of fluconazole versus amphotericin B for treatment of candidemia in non-neutropenic patients. Canadian Candidemia Study Group. AB - A randomized trial was conducted to compare the efficacy and safety of fluconazole versus that of amphotericin B in the treatment of candidemia in non neutropenic adults. Enrollment was stratified by disease severity (APACHE II score). Patients were randomized (1:1) to receive amphotericin B 0.6 mg/kg/day (cumulative dose 8 mg/kg) or fluconazole 800 mg intravenous loading dose, then 400 mg daily for four weeks (intravenous for at least 10 days). Patients were monitored for six months. A total of 106 patients were enrolled. A protocol amendment implemented midway through the trial required patients to be removed from the study and treated with amphotericin B if species identification indicated candidemia due to Candida glabrata or Candida krusei. Baseline characteristics were similar for the two groups; 103 patients (fluconazole, 50; amphotericin B, 53) met the major enrollment criteria. The intention-to-treat analysis indicated successful therapy in 50% of fluconazole recipients compared to 58% of the amphotericin B group (p = 0.39; one-sided 95% CI, -8 to 24%). The efficacy analysis included 84 patients (fluconazole, 42; amphotericin B, 42); successful outcomes were observed in 57% and 62% of cases in the fluconazole and amphotericin B groups, respectively (p = 0.66: one-sided 95% CI, -12 to 22%). The mortality at day 14 for the fluconazole group was 26% and for the amphotericin B group 21% (p = 0.52; chi-square test) and remained similar throughout the course of follow-up, Drug-related adverse events were more frequent with amphotericin B than with fluconazole and prompted switching of therapy for two (4%) and zero cases, respectively. Fluconazole and amphotericin B were associated with similar clinical response rates and survival in the treatment of candidemia among non neutropenic patients; however, drug-related adverse events were more frequent with amphotericin B. PMID- 9228473 TI - Semiquantitative polymerase chain reaction enzyme immunoassay for diagnosis of disseminated candidiasis. AB - A polymerase chain reaction enzyme immunoassay (PCR-EIA) was developed for the semiquantitative of circulating candidal DNA in disseminated candidiasis due to Candida albicans. Polymerase chain reaction was based on primers from the internal transcribed ribosomal region. Binding of the product to a streptavidin coated microtitration plate was mediated by a biotinylated capture probe. The product was digoxigenylated during PCR; this was the tag to which antibody was bound in the subsequent EIA. The optical density (OD) endpoint was < 0.1 in 15 sera from patients with no evidence of candidal infection (group 1) and in 13 of the 16 sera from colonized patients (group 2); it was > 0.1 in the other three sera from group 2 blood culture-negative patients who required intravenous amphotericin B for cure. The OD was positive in 28 patients with disseminated candidiasis (group 3), defined as positive blood cultures and successful treatment with amphotericin B (n = 11), positive blood culture confirmed at autopsy (n = 11), or negative blood culture first proven at necropsy (n = 6). In patients from whom multiple samples were available, recovery correlated with an optical density of < 0.1 by day 4 in four patients and by day 13 in the rest. In the five patients with fatal outcome from whom multiple samples were available, the mean OD rose from 0.174 to 0.668. Samples seeded with Candida albicans blastoconidia demonstrated that on OD of 0.220 was equivalent to 10 cfu. Assay of the group 3 sera by a commercial antigen detection test gave a corresponding sensitivity of 60% which rose to 67.9% when an in-house reverse passive latex agglutination test was used. PMID- 9228474 TI - Staphylococcus aureus nasal carriage as a marker for subsequent staphylococcal infections in intensive care unit patients. AB - From January to December 1994, 752 consecutive patients admitted to intensive care units (ICU) for more than two days were studied prospectively for Staphylococcus aureus colonization and infection. Nasal swabs were obtained at admission and weekly during the ICU stay. At ICU admission 166 patients (22.1%) were Staphylococcus aureus nasal carriers, while 586 were free of nasal colonization. Of the 166 nasal carriers, 163 harbored methicillin-sensitive Staphylococcus aureus (MSSA) and three methicillin-resistant Staphylococcus aureus (MRSA). During the ICU stay 24 of the 586 noncolonized patients became nasal carriers (11 MSSA and 13 MRSA), and one nasal carrier initially colonized by MSSA was reconlonized by MRSA. Staphylococcal infections were documented in 51 (6.8%) of the total 752 patients. After 14 days of ICU stay, the probability of developing staphylococcal infections was significantly higher for those patients who were nasal carriers at ICU admission than for those found to be initially negative (relative risk 59.6, 95% CI 20.37-184.32; p < 0.0001). In patients with ICU-acquired nasal colonization, most infections were documented prior to or at the time of the detection of the nasal colonization; thus, in this group of patients nasal carriage showed a lower predictive value for subsequent Staphylococcus aureus infections that that described classically. Paired isolates of nasal colonizing and clinical strains were studied by pulsed-field gel electrophoresis (PFGE) and mecA polymorphism analysis in 30 patients; identity was demonstrated in all but two patients. The results suggest that, outside the setting of an outbreak of MRSA, the detection of Staphylococcus aureus nasal carriers on admission may be particularly useful in identifying those patients who are at high risk for developing staphylococcal infections during their ICU stay. PMID- 9228475 TI - Detection of fastidious mycobacteria in human intestines by the polymerase chain reaction. AB - The aim of this study was to determine whether difficult-to-grow mycobacteria are present in human intestines. Intestinal tissue samples were subjected to both mycobacterial culture and a polymerase chain reaction (PCR) assay. After detection by PCR, species identity was determined by hybridizing the amplified 16S rRNA gene fragments with species-specific oligonucleotides. Intestinal biopsies from 63 patients with noninflammatory bowel diseases (n = 22), Crohn's disease (n = 31), or ulcerative colitis (n = 10) were analyzed. Culture and PCR revealed mycobacteria in four (6%) and 25 (40%) samples, respectively. Samples positive by PCR were negative with all probes specific to nine common cultivable species but were positive with Mycobacterium genavense-specific probe in 68% of cases. Mycobacterial isolates were identified as Mycobacterium gordonae and Mycobacterium chelonae. Findings were similar in Crohn's disease samples compared to non-Chron's disease samples. This study shows that difficult-to-grow mycobacteria can be detected by PCR in large and similar proportions of inflamed intestinal tissue from patients with inflammatory bowel disease and intestinal tissue that appears normal from patients with noninflammatory bowel disease. PMID- 9228476 TI - Tolerance and efficacy of Amphotericin B inhalations for prevention of invasive pulmonary aspergillosis in haematological patients. AB - The tolerance of aerosolised amphotericin B as prophylaxis against invasive pulmonary aspergillosis was investigated in 61 granulocytopenic periods in 42 patients treated for a haematologic malignancy. Each patient was to receive amphotericin B in doses escalating to 10 mg three times daily (t.i.d.), but only 20 (48%) patients managed to complete the scheduled regimen. One patient tolerated the full dose initially, but had to discontinue treatment when dyspnea developed as a result of pneumonia and acute respiratory distress. Another 22 patients (52%) experienced side effects, including eight (19%) who reported mild coughing and dyspnea but who tolerated the full dose and three (7%) patients whose dose was reduced to 5 mg t.i.d. Another six (14%) patients could tolerate only 5 mg t.i.d., and five (12%) others stopped treatment because of intolerance. Elderly patients (p < 0.05) and those with a history of chronic pulmonary obstructive disease (p = 0.09) were more likely to develop side effects during inhalation. Twelve (28%) patients developed proven of possible invasive fungal infections, but no correlation was established between infection and the total amount of amphotericin B inhaled. Inhalation of aerosolised amphotericin B is poorly tolerated and does not appear useful in preventing invasive pulmonary aspergillosis in granulocytopenic patients. PMID- 9228477 TI - Species-specific identification of microsporidia in stool and intestinal biopsy specimens by the polymerase chain reaction. AB - In view of the increasing number of cases of human microsporidiosis, simple and rapid methods for clear identification of microsporidian parasites to the species level are required. In the present study, the polymerase chain reaction (PCR) was used for species-specific detection of Encephalitozoon cuniculi. Encephalitozoon hellem, Encephalitozoon (Septata) intestinalis, and Enterocytozoon bieneusi in both tissue and stool. Using stool specimens and intestinal biopsies of patients infected with Enterocytozoon bieneusi (n = 9), Encephalitozoon spp. (n = 2), and Encephalitozoon intestinalis (n = 1) as well as stool spiked with spores of Encephalitozoon cuniculi and Encephalitozoon hellem and tissue cultures of Encephalitozoon cuniculi and Encephalitozoon hellem, three procedures were developed to produce PCR-ready DNA directly from the samples. Specific detection of microsporidian pathogens was achieved in the first PCR. The subsequent nested PCR permitted species determination and verified the first PCR products. Without exception, the PCR assay confirmed electron microscopic detection of Enterocytozoon bieneusi and Encephalitozoon intestinalis in stool specimens and their corresponding biopsies and in spiked stool samples and tissue cultures infected with Encephalitozoon cuniculi and Encephalitozoon hellem. Moreover, identification of Encephalitozoon spp. could be specified as Encephalitozoon intestinalis. Whereas standard methods such as light and transmission electron microscopy may lack sensitivity or require more time and special equipment, the PCR procedure described facilitates species-specific identification of microsporidian parasites in stool, biopsies, and, probably, other samples in about five hours. PMID- 9228478 TI - Invasive Haemophilus influenzae in the Republic of Ireland. AB - Prior to the general availability of Haemophilus influenzae type b vaccine in the Republic of Ireland, a two-year study of the epidemiology of invasive Haemophilus influenzae disease was carried out. Of 137 invasive strains of Haemophilus influenzae examined in a central laboratory, 94.2% were serotype b and 90.5% were biotype I. Seventeen percent of serotype b strains produced beta-lactamase, and 2.3% were resistant to both ampicillin and chloramphenicol. The majority of serotype b strains were electrophoretic types of the electrophoretic 12 clone family, principally 12.5. Meningitis was the most common infection caused by serotype b. The study data extend the current knowledge of strains of Haemophilus influenzae causing invasive disease in the Republic of Ireland. PMID- 9228479 TI - Culture of Helicobacter pylori from gastric biopsies transported in biopsy urease test tubes. AB - Gastric biopsy specimens of 57 consecutively observed dyspeptic patients were studied for the presence of Helicobacter pylori by histological examination, biopsy urease test (BUT) and culture. For culture, biopsy samples were transported in both Stuart media and BUT tubes. All 15 isolates could be cultured from both Stuart and BUT tubes. Thus, if the main reason for culture of Helicobacter pylori is for antimicrobial susceptibility testing, only positive BUT tubes need to be submitted. This would reduce both the expense and the number of biopsies needed. PMID- 9228480 TI - Nocardia otitidiscaviarum infection of a traumatic skin wound. AB - A case of primary Nocardia otitidiscaviarum infection of a skin wound over an open fracture in a previously healthy adult who suffered multiple trauma in a car accident is reported. The organism was identified in cultures of pus specimens from the infected wound. The case demonstrated the difficulties of testing susceptibility of nocardiae in vitro and the necessity of prolonged antibiotic treatment. The prevalence of nocardial infections is underestimated, highlighting the need for adequate documentation of such infections and compilation of the information by public health authorities. PMID- 9228481 TI - Pulmonary disposition of vancomycin in critically ill patients. AB - Vancomycin penetration in epithelium lining fluid was studied in ten mechanically ventilated patients with methicillin-resistant Staphylococcus aureus pneumonia 24 hours after the onset of treatment. Vancomycin was given intravenously at a daily dose of 30 mg/kg. Vancomycin levels were detectable in four patients (range, 1 2.77 micrograms/ml). Concordance between high plasma concentrations (> 20 micrograms/ml) and detectable vancomycin levels in epithelium lining fluid was noted. These results suggest that the pulmonary disposition of vancomycin remains low for most patients 24 h after the onset of treatment compared with the minimum inhibitory concentrations for most gram-positive organisms. One therapeutic goal of vancomycin treatment could be to obtain through plasma levels of 20 micrograms/ml. Further studies are required to determine the clinical relevance of these observations. PMID- 9228482 TI - Etiology and response to antibiotic therapy of community-acquired pneumonia in French children. AB - The aim of this study was to determine the etiologic agents associated with community-acquired pneumonia in 104 French children ages 18 months to 13 years. Potential respiratory pathogens were identified in 87 (85%) cases; these included respiratory syncytial virus in ten, other viruses in 20, Streptococcus pneumoniae in 14 and Mycoplasma pneumoniae (diagnosed by serologic procedures) in 43. Of 32 patients with Mycoplasma pneumoniae infection who were initially treated with beta-lactam antibiotics, 30 failed treatment. Recovery from mycoplasma infection occurred rapidly in patients treated with macrolide antibiotics (which included spiramycin in 31 patients, josamycin in 7, and erythromycin in 3); however, cough persisted in 12 patients for one month. The high frequency of Mycoplasma pneumoniae in children over 18 months of age must be considered when selecting an antibiotic for initial therapy. PMID- 9228483 TI - Comparison of agar dilution, broth dilution, disk diffusion, and the E-test for susceptibility testing of penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae. AB - An evaluation to determine the optimal methods for the in vitro susceptibility testing of 41 clinical isolates and the ATCC 49619 strain of Streptococcus pneumoniae to penicillin was undertaken. No very major or major interpretive errors were observed with the following test methods and media: agar dilution using either Mueller-Hinton medium with lysed horse blood or Haemophilus test medium; broth dilution using cation-adjusted Mueller-Hinton medium with lysed horse blood, Haemophilus test medium, or Todd-Hewitt medium; and the epsilo-meter test (E-test) using agar containing Mueller-Hinton medium and 5% sheep blood. The disk diffusion method using agar containing Mueller-Hinton medium and 5% sheep blood agar was an effective screening method, requiring confirmation by a dilution susceptibility test method. PMID- 9228484 TI - Nonradioactive single-strand conformation polymorphism analysis for detection of fluoroquinolone resistance in mycobacteria. AB - A simple, rapid, and nonradioactive method for routine detection of fluoroquinolone resistance in mycobacteria is described. A single-strand conformation polymorphism (SSCP) methodology, based on the use of mini-gels and silver staining of DNA, was optimized for the analysis of denatured DNA products obtained by polymerase chain reaction (PCR) from the gyrA gene involved in fluoroquinolone resistance in mycobacteria. The method was successfully applied to fluoroquinolone-susceptible and -resistant laboratory strains of Mycobacterium smegmatis and to clinical strains of Mycobacterium tuberculosis isolated from patients who developed resistance during the course of fluoroquinolone treatment. PMID- 9228485 TI - Transmission of methicillin-resistant staphylococcus aureus within a household. PMID- 9228486 TI - Garderella vaginalis bacteremia in an adult male. PMID- 9228487 TI - Strongyloides stercoralis infection in a patient with Crohn's disease. PMID- 9228488 TI - Gestational diabetes and preterm labour: is glycaemic control a contributing factor?. AB - OBJECTIVES: (1) to evaluate the incidence of preterm delivery in patients with gestational diabetes mellitus; (2) to determine the association between glycaemic control and preterm delivery in these patients. STUDY DESIGN: (1) The incidence of spontaneous preterm singleton deliveries was determined in 550 intensively treated patients with gestational diabetes mellitus. A total of 14 552 consecutive patients without gestational diabetes mellitus who delivered during the same interval served as a control population; (2) Glycaemic profiles (i.e., mean blood glucose, percent of hypoglycaemic [ < 60 mg/dl] and hyperglycaemic [ > 120 mg/dl] episodes) were compared in 34 patients with gestational diabetes mellitus who delivered preterm, and 68 matched controls with gestational diabetes mellitus who delivered at term. RESULTS: (1) The incidence of preterm delivery in gestational diabetics was similar to that found in the non-diabetic population (6.2% vs. 6.5%, respectively, P = 0.82; confidence limits: 0.65, 1.36); (2) women with gestational diabetes mellitus who delivered at term, or preterm had similar glycaemic profiles for both the entire treatment period and the week preceding delivery. CONCLUSIONS: (1) There is no increased risk for preterm delivery in intensively-treated gestational diabetes mellitus patients; (2) In a population such as this women with gestational diabetes mellitus who deliver preterm cannot be characterised by their glycaemic profile. PMID- 9228489 TI - Neurological findings in neonates with low Apgar in Zimbabwe. AB - OBJECTIVES: Document neurological condition of African neonates with a low apgar score. SETTING: Mpilo Hospital, Bulawayo, Zimbabwe. SUBJECTS: 165 babies with an Apgar score of 5 or less at 5 min. METHODS: Neurological examination at term age according to Prechtl. Babies were classified as normal, suspect or abnormal and compared with two reference groups, one from Groningen, the Netherlands and one from Grenada in the Caribbean. RESULTS: A higher number of Zimbabwean babies were delivered by Caesarean section compared to the Groningen group (P < 0.001). Babies delivered by vacuum extraction scored significantly lower compared to babies delivered by Caesarean section (P < 0.003). Twenty abnormal signs derived from the neonatal neurological examination proved to be predictive on the total optimality score (P < 0.001). The number of infants who were classified as abnormal was higher in the Zimbabwean population (P < 0.01). CONCLUSION: The selected abnormal signs derived from the neonatal neurological examination proved to be highly predictive on the neurological condition. The neonatal morbidity in Zimbabwean neonates with a low Apgar score was higher when compared with two reference groups from Groningen and Grenada. PMID- 9228491 TI - Genetic factors in the aetiology of gastroschisis: a case report. AB - Isolated gastroschisis in two children of unrelated mothers but with the same father is described. There was no evidence of consanguinity, abnormal karyotype or a specific teratogen in either case. Genetic factors may be involved in the aetiology of some cases of gastroschisis. PMID- 9228490 TI - Plasma concentrations of beta-endorphin and adrenocorticotropic hormone in women with and without childbirth preparation. AB - We studied plasma concentrations of beta-endorphin (beta-EP) and adrenocorticotropic hormone (ACTH) during dilation, expulsion and immediate puerperium in 47 primiparous women with an uneventful pregnancy and spontaneous vaginal delivery. Twenty-five women had received childbirth preparation with the Lamaze method, and 22 had received no preparation. Mean concentrations of beta-EP from the beginning of labor until puerperium were higher in women who had received preparation, but there was no significant difference between the two groups. When behavior during labor was evaluated regardless of which group the patient was assigned to, women whose behavior was unsatisfactory has significantly higher concentrations of ACTH at all times during childbirth. We discuss the role of childbirth preparation as a way to enhance beta-EP secretion. Levels of ACTH, on the other hand, appear to be more closely related with behavior during labor, regardless of whether the mother received preparation. PMID- 9228492 TI - Role of antepartum computerized fetal heart rate analysis in the prediction of fetal distress during labor. AB - OBJECTIVE: To ascertain the diagnostic ability of a computerized fetal heart rate (FHR) analysis system in the identification of patients at risk of fetal distress in labor. STUDY DESIGN: Five-hundred and seventy-seven healthy term pregnancies were enrolled in a prospective, cross-sectional study and subdivided into two groups, with (n = 90) or without (n = 487) fetal distress in labor. Computerized FHR recordings were performed periodically and regression analysis was performed in a univariate and multivariate way to assess the ability of the last FHR recordings in the prediction of subsequent fetal distress. RESULTS: The two groups showed a significant difference in almost all of the FHR parameters studied. The multivariate analysis showed that only the FHR baseline and the percentage of small acceleration were independently and significantly related to the labor outcome. The combination of these two parameters reaches a sensitivity of 45%, specificity of 91%, positive predictive value of 48% and negative predictive value of 90%, with an overall accuracy of 84%. CONCLUSIONS: The increase in FHR baseline and in small FHR accelerations can be major factors in the prediction of subsequent fetal distress in healthy term fetuses. PMID- 9228493 TI - Transvaginal surgery for uterine scar dehiscence. AB - Asymptomatic uterine rupture, usually discovered during routine uterine examination because of a pre-existing uterine scar, it treated by techniques that include suturing the dehiscence via abdominal access, total or subtotal hysterectomy or therapeutic non-intervention. The authors propose a transvaginal technique. PMID- 9228494 TI - The effect of nitric oxide on uterine and umbilical artery flow velocity waveform in pre-eclampsia. AB - We compared the flow velocity waveforms of uterine and umbilical arteries in normotensive and pre-eclamptic patients at mid-gestation. In a randomised controlled trial we tested the effects of isosorbide dinitrate (ISDN, nitric oxide donor) patch therapy on the flow velocity waveform of pre-eclamptic patients at mid-gestation. The resistance indices (RI) of human uterine and umbilical arteries were higher in pre-eclamptic patients compared to the normotensive patients. ISDN patch therapy significantly reduced the increased RI values of the umbilical artery in pre-eclamptic patients without any change in systemic blood pressures, but the RI values of the uterine artery were not significantly attenuated. The change of the umbilical artery might be due to the improvement of end-diastolic flow velocity. These results suggest that the feto placental circulation in pre-eclampsia, perhaps due to the disturbance of the endothelium-dependent vaso-relaxation system, and that ISDN therapy may improve the impaired endothelium dependent nitric oxide system. PMID- 9228495 TI - Myotonic dystrophy in pregnancy: a report of two cases within one family. AB - myotonic dystrophy, also called the Curschmann-Steinert syndrome, is an autosomal dominant inherited neuromuscular disorder characterized by progressive muscular dystrophy, muscle weakness and myotonia, which can affect both mother and child. Complications may arise during pregnancy, delivery, including anaesthetic problems, and in the neonatal period. During pregnancy hydramnion can be a first sign of the disease leading to premature labor and also muscle weakness and myotonia can aggravate complicating the course of delivery. The affected neonate may display severe hypotonia, facial diplegia and respiratory distress. The clinical diagnosis can be confirmed by direct DNA analysis in serum and in chorionvillus biopsy material. In this case report two sisters with myotonic dystrophy are described, their pregnancies, deliveries and the outcome of their affected babies. PMID- 9228496 TI - Twenty two weeks of transdermal estradiol increases sex hormone-binding globulin in surgical menopausal women. AB - OBJECTIVE: To compare the effects of continuous noncombined transdermal estradiol versus oral conjugated estrogen on serum sex hormone-binding globulin (SHBG) levels prior to and during the 10th and 22nd weeks of therapy in patients with surgical menopause. STUDY DESIGN: Open, comparative trial. Patients were consecutively assigned to three groups: group 1 (n = 18) received continuous transdermal estradiol (0.050 mg/day), group 2 (n = 18) continuous oral conjugated estrogens (0.625 mg/day), whereas group 3 (n = 15) received no treatment. Serum SHBG levels were determined before treatment and after 10 and 22 weeks of treatment. RESULTS: Serum SHBG increased significantly with oral conjugated estrogens at 10 (p < 0.01) and 22 weeks (p < 0.01) compared with baseline. With transdermal estrogens there was a much smaller increase of SHBG. At 22 weeks, this increase was significant compared with baseline (p < 0.05), but not compared with the control group (p > 0.05). CONCLUSION: Transdermal estrogen has no effect on SHBG, whereas oral conjugated estrogens causes considerable increase. PMID- 9228497 TI - Long-term follow-up on the treatment of endometriosis with the GnRH-agonist buserelinacetate. Long-term follow-up data (up to 98 months) of 42 patients with endometriosis who were treated with GnRH-agonist buserelinacetate (Suprecur), were evaluated in respect of recurrence of pain symptoms and pregnancy outcome. AB - OBJECTIVE: In our previous study, 119 patients with histologically confirmed endometriosis underwent a 'three-step' therapy between 1987 and 1989, where surgical removal of endometriosis was followed by a 6 month treatment with 3 x 300 microgram buserelinacetate daily intranasally and a second look laparoscopy or laparotomy with removal of residuals. Long-term follow-up data in respect of recurrence of symptoms and pregnancy outcome were investigated. STUDY-DESIGN: Long-term follow-up data of 42 out of 119 treated patients on the post-treatment effect were obtained using a special questionnaire. Recurrence of dysmenorrhea, dyspareunia and pelvic pain was defined as recurrence of disease. The follow-up period was up to 98 months with a median time of 82.5 months. RESULTS: Out of the 42 patients, 23 complained of infertility. Fourteen out of these 23 patients became pregnant during the follow-up period, resulting in 23 pregnancies with 18 newborns, 4 miscarriages and one ectopic pregnancy. Ten patients conceived spontaneously, stimulation program became necessary in the rest of patients. Twenty-eight of the 42 patients complained recurrence of symptoms with median first onset at 10.7 months. Improvement on quality of life and subjective conditions were reported by 30 patients. CONCLUSIONS: Our study suggests that the 'three-step' therapy of endometriosis with GnRH-agonist buserelinacetate leads to a significant improvement on the quality of life and well being in the majority of the patients and to a high pregnancy rate. This treatment represents a favourable approach in the management of endometriosis. PMID- 9228498 TI - Inhibin assays of ovarian cyst liquid obtained by needle aspiration may allow differential diagnosis between functional and organic cysts. AB - Radioimmunoassays of estradiol, CA125 and inhibin were carried out on ovarian cyst fluid samples. The samples were taken from ten women with functional cysts and 15 women with organic cysts. Statistical analysis shows that estradiol and inhibin assays allows satisfactory differential diagnosis between functional and organic cysts. However, the inhibin assay provides more precise results and appears to be a more reliable marker than estradiol. The differential diagnosis between the two types of cyst would indicate the use of coelioscopic surgery especially in the case of organic cysts, avoiding unnecessary risks for patients with functional cysts. PMID- 9228499 TI - Fluoxetine in the treatment of premenstrual syndrome. AB - BACKGROUND: Premenstrual syndrome (PMS) is defined as the disabling and cyclic occurrence of emotional and behavioral symptom complex during the latter half of the menstrual cycle. Although its etiology is unknown, it has been speculated that premenstrual syndrome is linked to a deficiency of central serotoninergic activity. METHOD: The study consisted of a double-blind, placebo controlled trial of fluoxetine at a dose of 20 mg/day or placebo for three menstrual cycles. The 440 women who appeared to meet the eligibility criteria were instructed to record the 'Calendar of Premenstrual Experiences' (CPE) scale for two complete menstrual cycles. Of 410 women who successfully completed two cycles of recording their symptoms daily only 35 met the criteria for PMS. These criteria included psychiatric interviews which were made before treatment. Thirty-five PMS patients were randomized into placebo or fluoxetine treatment groups. RESULTS: Our study suggests that fluoxetine at a dose of 20 mg per day was significantly superior to placebo in alleviating the symptoms of PMS. The most common side effects were gastrointestinal irritability (15%), insomnia (11%) and sexual dysfunction (8.5%). CONCLUSION: Fluoxetine is an effective and well-tolerated drug and appears to have considerable promise in treating a range of symptoms in women with PMS. PMID- 9228500 TI - Immunohistochemical analysis of glutathione S-transferase mu expression in ovarian tumors. AB - OBJECTIVE: To explore the association between Glutathione S-transferase (GST) -mu expression and clinicopathologic features in ovarian tumors. STUDY DESIGN: Immunohistochemical study was made to investigate GST-mu expression in diverse ovarian tumors. RESULTS: All 75 ovarian tumors expressed GST-mu. There was no significant association between GST-mu immunopositivity and clinicopathological features. In serous cystadenocarcinoma, however, patients with weak GST-mu expression survived longer than those with moderate or strong GST-mu expression. Three of the 8 tumors that had expressed weak GST-mu initially increased GST-mu expression after chemotherapy. CONCLUSION: GST-mu expression is common in ovarian tumors and malignant tumors expressed more GST-mu than benign tumors. GST-mu might play a major role in the development of drug resistance in certain ovarian tumors and could be a useful marker of natural resistance and their future outcome. PMID- 9228501 TI - Clomiphene citrate challenge test in the assessment of ovarian reserve before controlled ovarian hyperstimulation for intracytoplasmic sperm injection. AB - The objective of this study is to evaluate the performance of clomiphene citrate (CC) challenge test to predict diminished ovarian reserve before controlled ovarian hyperstimulation for intracytoplasmic sperm injection (ICSI). The 198 women who underwent the CC challenge test fulfilled the following criteria; over 35 years of age, removal of one ovary or previous ovarian surgery, the presence of ovarian endometrioma or previous poor response to ovarian hyperstimulation. Of the patients tested, 141 were found to have a normal CC challenge test while 57 had an abnormal result. The cancellation rate of the cycle with a poor response was significantly higher in women with an abnormal test (36.8%) than in those with a normal test (19.8%) (P < 0.05). The sensitivity of CC test for cycle cancellation was found to be 43% with a specificity of 76%, positive and negative predictive values of 37 and 80%, respectively. The estradiol values on hCG day, the number of retrieved oocytes and metaphase II oocytes and the rate of transfer cycles were significantly lower in females with an abnormal test. Women with normal test results had higher pregnancy rates per embryo transfer than those with abnormal test results (21.5 vs. 13.3%) and the predictive value of an abnormal test for failing to conceive was 93% (53/57) with a sensitivity of 31%, specificity of 84% and negative predictive value of 15.6%. Of 57 women with an abnormal test result, 25 (43.8%) were abnormal due only to an elevated day 10 or 11 value of FSH, which could not be detected using only basal FSH screening. In this group, the cancellation rate (48 vs. 19.8%, P < 0.01), the rate of transfer cycles (48 vs. 72.3%, P < 0.05) and the mean number of retrieved oocytes (4.9 +/- 2.5 vs. 6.4 +/- 3.1, P < 0.01) were all significantly different from normal test group. Although the rate of pregnancies per started cycle (8 vs. 15.6%) did not show a statistically significant difference, this is most probably due to the low number of patients. In conclusion, an abnormal CC challenge test is a good predictor of diminished ovarian reserve and it is better than a basal FSH concentration on day 3. It provides valuable information for both patients as to their chances of achieving a pregnancy and also for the medical team deciding on options for stimulation protocols. PMID- 9228502 TI - No effects of human relaxin on the active and passive biomechanical properties of isolated cervical specimens from nonpregnant women. AB - OBJECTIVE: To evaluate the effect of human relaxin (hRLX-2) on the active and passive biomechanical properties of cervical tissue in vitro. MATERIAL: Cervical samples were obtained from the middle part of the cervix in 22 nonpregnant women undergoing hysterectomy. METHODS: The effect of hRLX-2 (10(-7) M) on the active biomechanical properties was studied on vasopressin (10(-8) M) induced smooth muscle contractions in an organ bath model. The effect on the passive biomechanical properties were studied after incubation of the strips for 48 h with hRLX-2 (10(-8) M and 10(-9) M). Subsequently, the specimens were stretched in a material testing machine until they broke. The load applied and the elongation were simultaneously recorded and the results translated into stress strain curves. RESULTS: hRLX-2 did not influence the vasopressin-induced contractility of cervical strips from nonpregnant women in this study. No synergistic effect of progesterone could be demonstrated. The passive biomechanical properties (tensile strength, extensibility, stiffness of failure energy) did not change significantly after relaxin incubation. The results obtained in vitro do not suggest an important physiological effect of relaxin on the human nonpregnant cervix. PMID- 9228503 TI - Hypermobility in two Dutch school populations. AB - OBJECTIVE: To determine the presence of hypermobility and differences between females and males in a Dutch population. STUDY DESIGN: Joint mobility was measured in a primary and a secondary school population. Beighton and Biro measurements were used. The data were evaluated statistically. RESULTS: Using the Beighton score, 15.5% of group I (n = 252; 4-13 years) and 13.4% of group II (n = 658; 12-17 years) were hypermobile. Hypermobility was found more in females than in males, the difference being significant in the older group. Overall, hypermobility did not significantly diminish with ageing, although the individual joints did not show a significant decrease in mobility with ageing. Hypermobility was significantly more pronounced at the non-dominant body side in both groups. The Quetelet-index did not show a significant relation to hypermobility. CONCLUSION: Hypermobility was found more in females than in males, with a trend of decrease of hypermobility with ageing. The non-dominant body side proved to be more hypermobile and the Quetelet-index did not show a relation to hypermobility. Beighton's measurements proved best, since Biro considers the two body sides being equal. PMID- 9228504 TI - Case report: denovo inherited 18p deletion in a mother-fetus pair with extremely variable expression, confirmed by fluorescence in situ hybridization (FISH) analysis. AB - Denovo deletions of 18p without other associated rearrangement are uncommon. For such a deletion to profoundly affect the fetus of a near normal phenotypic carrier would be rarer. We present such a case in which the chance of a cryptic rearrangement was ruled out by fluorescence in situ hybridization (FISH) analysis. Possible explanations for wide variations in clinical expression are discussed. PMID- 9228506 TI - Lumbosacral plexus compression by fetus: an unusual cause of radiculopathy during teenage pregnancy. AB - A case is reported of a lumbosacral plexus compression by the fetus in a young 34 weeks pregnant woman, who had low-back pain and progressive muscular weakness of the leg. Neurological examination showed a grade IV motor weakness of the iliopsoas, quadriceps femoris and biceps femoris muscles. Mechanical stretch manoeuvers were negative. Electromyography revealed denervation activity in L4 and L5 muscles. Lumbosacral plexus radiculopathy was diagnosed. Although fetal compression appears to be an uncommon cause of lumbosacral radiculopathy during teenage pregnancy, both neurosurgeons and obstetricians should be aware of the possibility. PMID- 9228505 TI - Ultrasonography and magnetic resonance imaging findings in a patient with an unruptured interstitial pregnancy. AB - We report a case of an unruptured interstitial pregnancy, which showed characteristic ultrasonographic and magnetic resonance imaging (MRI) findings. Color and pulsed Doppler sonography may facilitate early diagnosis of interstitial pregnancy. MRI may play some role in the diagnosis of this entity when ultrasound studies are insufficient or equivocal. PMID- 9228507 TI - Basic science and clinical utility of biochemical markers of bone turnover--a Congress report. PMID- 9228508 TI - No apparent benefit of liquid formula diet in NIDDM. AB - We have studied the impact of liquid diets formulated for complete or supplemental enteral nutrition of type II, non insulin-dependent (NIDDM) diabetics on carbohydrate homeostasis. To achieve this, liquid formula tolerance tests were performed in NIDDM patients under an oral treatment regimen with a combination of diet plus glibenclamide (7 men, 3 women, age: 56 +/- 11 years; mean body mass index of 26.2 +/- 3.6 kg/m2). After an overnight fast, each patient received the usual morning medication and, thereafter, ingested formula diet in randomized order with 10 day intervals between tests. 500 ml were administered of either a standard liquid diet (Biosorb Sonde), a fibre containing diet (Biosorb Plus Sonde), or a carbohydrate modified, fructose containing "diabetes" diet (Fresubin Diabetes) (carbohydrate contents of approximately equal to 60 g, respectively). Blood samples were collected over 180 min. Considering minor variations in the nutritional values of the diets, IR-insulin, IR-C peptide, IR-glucose-dependent insulinotropic polypeptide (GIP), and IR-glucagon like peptide. 1 (GLP-1) in plasma did not significantly differ between the study groups. After ingestion of Biosorb Sonde area under the curve glucose was greater than that seen after uptake of fibre containing or carbohydrate modified, i.e. fructose containing "liquid diabetes diets". All diets challenged the entero insular axis in non-insulin dependent diabetics to a comparable extent. This data does not support the contention to employ special "diabetes" formulas for enteral nutrition of patients with NIDDM. PMID- 9228509 TI - Comparison of the time-action profiles of U40- and U100-regular human insulin and the rapid-acting insulin analogue B28 Asp. AB - It is well known that rapid-acting insulin analogues like insulin aspart (B28Asp) show a faster onset and a shorter duration of action than currently available U100 soluble insulin preparations. Since this effect is mainly due to a lower concentration of slow-absorbable hexamers, the metabolic profile of human insulin in lower concentration (e.g. U40 insulin) might be more similar to that of insulin aspart. Therefore, we compared the pharmacodynamic and pharmacokinetic properties of U40 soluble insulin with insulin aspart (U100) and with U100 human insulin. Eight healthy volunteers received on different study days s.c. injection of insulin aspart and U40 insulin (0.2 U/kg body weight) under euglycaemic clamp conditions (blood glucose 5 mmol/l, basal i.v. insulin infusion 0.15 mU/kg/min). In a second study, U40 and U100 soluble insulin was administered to 9 other volunteers under similar conditions. No significant differences were observed between the summary measures of U40 and U100 insulin. Insulin aspart showed a faster onset and a shorter duration of action than U40 insulin. The metabolic activity of human insulin in concentrations of U40 and U100 is comparable. Subcutaneous injection of the insulin analogue insulin aspart leads to a faster onset and a shorter duration of action even in comparison to U40 insulin. PMID- 9228510 TI - Diabetic microangiopathy and urinary glycosaminoglycans. AB - Alterations in the metabolism of glycosaminoglycans (GAG) may play a role in the pathogenesis of diabetic-associated microangiopathy. Consequently, the relationship between diabetic nephropathy and retinopathy and urinary GAG distribution was assessed in 96 IDDM patients in comparison to 103 healthy controls. GAG concentration in 24h urine samples was determined by precipitation with cetylpyridinium chloride and potassium acetate in ethanol followed by a colorimetric test with carbazole. A marked difference (P = 0.0008) in urinary GAG excretion between patients (24.3 +/- 1.5 mg/24 h, mean +/- SEM) and controls (16.2 +/- 0.75 mg/24 h) could be detected. In patients with IDDM of longer duration, GAG excretion was increased (< or = 10 yr: 20.8 +/- 2.1 vs > 10 yr: 27.4 +/- 2.1 mg/24 h; P = 0.03). Furthermore, IDDM patients with class 4 nephropathy and retinopathy exhibited a markedly higher GAG excretion compared to those without nephropathy (33.1 +/- 3.0 vs 22.6 +/- 1.7 mg/24 h, P = 0.005) or retinopathy (29.7 +/- 2.8 vs 21.2 +/- 1.7 mg/24 h, P = 0.009). An increased urinary GAG concentration was detected in IDDM patients with albuminuria (> 300 mg/24 h: 29.9 +/- 3.3 vs < 30 mg/24 h: 23.0 +/- 1.7 mg/24 h; P = 0.048), proteinuria (> 0.5 g/24 h: 30.3 +/- 3.7 vs < 0.05 g/24 h: 22.7 +/- 1.6 mg/24 h) and in patients with augmented serum creatinine in comparison to those with normal values (> 0.12 mg/L: 34.9 +/- 2.3 vs < 0.12 mg/L: 22.4 +/- 1.6 mg/24 h; P = 0.01). The results demonstrate a close relationship renal GAG excretion and the presence of microangiopathy in IDDM patients. PMID- 9228511 TI - Improved islet isolation by 10% albumin does not influence graft angiogenesis and vascularization. AB - Addition of 10% albumin to the digestion medium has been suggested to enhance yield and integrity of harvested islets by inhibition of proteolytic activities and to improve endocrine function early after transplantation. The aim of this study was to evaluate in vivo by means of intravital fluorescence microscopy whether this rapid reversal of hyperglycemia after transplantation is due to improved graft vascularization. Pancreatic islets were isolated from Syrian golden hamsters by collagenase digestion using either solely Hank's balanced salt solution (HBSS) or HBSS supplemented with 10% human serum albumin. Islets were then transplanted into the dorsal skinfold chamber of syngeneic animals (control: N = 8 animals, n = 50 islets; albumin: N = 7, n = 41). The grafts' microvasculature was analysed on days 6, 10, and 14 after transplantation. Immunohistochemical staining for insulin was performed at the end of the microscopic observation period. Islet isolation with albumin supplementation did not increase islet yield. However, photomicroscopic analysis suggested a beneficial effect on the isolation process with improved islet integrity and prevention of outer margin irregularities, in particular in large islets. Analysis of revascularization 6 days after transplantation revealed in the control group a functional capillary density (FCD) of 477 +/- 47 cm-1. On day 10 FCD increased to 680 +/- 42 cm-1 with no further changes on day 14, indicating complete revascularization. Islets in the albumin group demonstrated a comparable FCD of 598 +/- 49 cm-1 on day 10 and complete revascularization on day 14 (655 +/ 45 cm-1). The angio-architecture of the islets was found similar in both groups, presenting with a glomerulum-like capillary network, comparable to that of pancreatic islets in situ. We conclude that the addition of 10% serum albumin to the collagenase digestion medium improves the preservation of the structural integrity of isolated pancreatic islets, however, does not influence the process of graft vascularization. Thus, improved early graft function may rather be due to superior preservation of islet cell integrity and function. PMID- 9228512 TI - High levels of circulating adrenomedullin in severe illness: correlation with C reactive protein and evidence against the adrenal medulla as site of origin. AB - Adrenomedullin (AM) is a novel vasorelaxing peptide which was originally isolated from the extracts of human pheochromocytoma. It is produced by a number of organs among which the adrenal gland exhibits by far the highest concentrations. The peptide circulates in blood and its plasma levels have been reported to be increased in several diseases such as renal failure and sepsis. In the present study plasma concentrations of AM were measured in various forms of severe illness and compared to clinical and biochemical parameters in order to gain an insight into the factors controlling the plasma levels of this peptide. The highest concentrations of AM were found in patients with sepsis (344.4 +/- 60.4 pg/ml, n = 16) who exhibited up to 12-fold higher levels than a group of healthy subjects (74.1 +/- 4.1 pg/ml, n = 20). Markedly elevated levels were also measured in hemorrhagic (250.1 +/- 37.9 pg/ml, n = 9) and cardiogenic (216.2 +/- 29.4 pg/ml, n = 7) shock as well as in patients with cancer of the gastrointestinal tract (155.6 +/- 32.5 pg/ml, n = 11) or the lungs (146.5 +/- 19.1 pg/ml, n = 22). Plasma AM levels were positively correlated with serum creatinine concentrations in shock (r = 0.06, p < 0.001) and with C-reactive protein levels in patients with cancer (r = 0.64, p < 0.001) or sepsis (r = 0.63, p < 0.01). In order to examine the potential role of the adrenal gland as a site of AM release, hypoglycemia was induced in a group of healthy volunteers by graded infusion of insulin. Despite a more than 20-fold increase in plasma adrenalin indicating maximal stimulation of the adrenal medulla, no significant alterations of the plasma AM levels were observed. The study demonstrates that not only sepsis but also various forms of cancer and shock are associated with high levels of circulating AM. The correlation with C-reactive protein levels suggests a role of cytokines in mediating the elevations in plasma AM observed in sepsis and cancer. Reduced clearance of the peptide by the kidneys may be one of the mechanisms involved in the accumulation of AM in shock. The adrenal gland appears not to be a major source for circulating AM. PMID- 9228513 TI - The use of salivary cortisol measurements for the non-invasive assessment of adrenal cortical function in guinea pigs. AB - For collection of saliva, cotton buds (Q-tips) were inserted into the guinea pig's cheek pouch, parallel with the cheek teeth. The recovery of saliva from the buds increased with the volume applied: whereas the recovery was 63% when 20 microliters were applied, it increased to 85% when 120 microliters were applied. The recovery of cortisol was closely related to the volume of saliva obtained by centrifugation (r = 0.99, p < 0.001). The mean value of salivary cortisol concentrations in untreated animals was 6.6 ng/ml, with relatively large variations across minutes and days within and between animals. Salivary cortisol was significantly increased if animals were singly caged, either in their familiar housing room or in an unfamiliar empty test room. In comparison to these changes, a much more pronounced increase of salivary cortisol occurred after the intramuscular (i.m.) administration of 20 IU ACTH: while the pretreatment value was 2.2 ng/ml, cortisol concentrations increased to 47 ng/ml (1 h), 72 ng/ml (2 h), 137 ng/ml (3 h) and 170 ng/ml (4 h), respectively. Similarly, i.m. administration of 2 IU insulin resulted in a significant increase of salivary cortisol (2 h: 37 ng/ml, 3 h: 24 ng/ml, 4 h: 25 ng/ml). The present study shows that the cortisol concentrations in the saliva of guinea pigs can be used as an index of adrenal cortical function in preference to the more commonly measured concentrations in the plasma. The advantages of the saliva method are: Firstly, cortisol values reflect the biologically active, unbound fraction in the plasma and are thus less affected by concentration changes of the corticosteroid-binding globulin. Secondly, saliva is easy to collect and the collection method is non invasive; thus, no handling- or stress-induced changes of the adrenal gland occur. Thirdly, the ease of collection facilitates investigations which require frequent sampling. PMID- 9228514 TI - Epididymal and sex accessory gland secretions in transfusion-dependent beta thalassemic patients: evidence of an impaired prostatic function. AB - Neutral alpha-glucosidase levels as epididymal marker, fructose levels as vesicular marker, zinc, citric acid and prostate specific antigen levels as prostatic markers were measured in the seminal plasma of eight transfusion dependent beta-thalassemic patients in order to study epididymal and sex accessory gland secretions (eighteen subjects served as controls). FSH and LH as well as total and free testosterone were detected displaying unaltered serum values. Ejaculate of patients showed normal sperm count and low sperm motility, in the meantime seminal plasma exhibited unaltered both neutral alpha-glucosidase and fructose values but low levels of zinc, citric acid and prostate specific antigen were noticed as well. These data suggest an impaired prostatic secretion in the thalassemic patients studied. A local iron toxicity on the prostatic tissue could be supported by the decrease of its specific markers observed only in the subgroup of patients with high ferritin serum levels. PMID- 9228515 TI - Extracellular monoamines and their metabolites in the mediobasal hypothalamus- median eminence of anestrous and estrous ewes during CRF treatment. AB - In order to clarify the effect of exogenous corticotropin-releasing factor (CRF) on catecholaminergic and serotoninergic system activity in the mediobasal hypothalamus-median eminence (MBH-ME) of ewes the changes in extracellular levels of noradrenaline (NA) and serotonin (5HT), and main metabolites of monoamines, 4 hydroxy-3-methoxyphenylglycol (MHPG), 3,4-dihydroxy-phenylacetic acid (DOPAC), homovanilic acid (HVA), and 5-hydroxy-indolo-3-acetic acid (5-HIAA) were quantified in the perfusates collected from MBH-ME. NA, 5-HT and monoamine metabolites in the perfusates were analyzed using high performance liquid chromatography with electrochemical detection. CRF induced a rise in extracellular concentration of NA and 5-HT only in the estrous ewes prior to a preovulatory LH surge. CRF treatment caused a heterogenous effect on extra cellular concentrations of 5-HT in ewes during the preovulatory LH surge. Except for DOPAC and HVA in some estrous ewes during the preovulatory LH surge, CRF caused an increase in monoamine metabolites levels in the MBH-ME in anestrous and estrous animals. These results indicate that CRF facilities NA release in the MBH ME during the presurge LH period in ewes, and that CRF increases metabolic activities of the monoaminergic systems in this structure in the anestrous and estrous ewes, except dopaminergic system in the ewes during the preovulatory LH surge. It is suggested that: 1) the responses of monoaminergic systems activity in the MBH-ME to CRF in large degree is dependent upon physiological state of ewes and 2) in some endocrinological phases CRF may affect LHRH and other hypothalamic hormone secretion indirectly by altering monoaminergic system activity in the MBH-ME. PMID- 9228516 TI - Adrenal functions in patients with sepsis. AB - The basal cortisol level and cortisol response to ACTH stimulation test were assessed in patients with sepsis, the results being compared to a control group of 30 healthy persons. The study group included 49 patients with sepsis and 30 healthy subjects as a control group. The mean age in the study group was 42.6 +/- 18.7 years and 41.4 +/- 12.1 years in the control group. Fifteen of the 49 (30.6%) patients had hospital-acquired and 34 (69.4%) patients community-acquired sepsis. Etiological agent was isolated in 35 (71.4%) patients (57.1% gram negative bacteria and 34.3% gram positive bacteria, plus 8.6% polymicrobial). Fourteen of 49 (28.6%) patients died. Mean basal cortisol level was 597.1 +/- 304.6 nmol/l (range 217.8-1667.9) in the study group and 460.2 +/- 180.8 nmol/l (range 253.6-988.9) in the control group. Mean basal cortisol level in the study group was significantly higher than that of the control group (p < 0.05). Mean basal cortisol level was found to be 725.5 +/- 448.9 nmol/l in the patients who died and 545.8 +/- 210.9 nmol/l in the patients who recovered. The difference between the two groups was found to be significant (p < 0.05). ACTH stimulation test was performed in 43 of the patients and 30 healthy subjects. Cortisol response was significantly lower (mean 277.7 +/- 216.9 nmol/l) in the patients than that detected in the control group (mean 519.6 +/- 279.2) (p < 0.001). Mean cortisol response in the patients who died was 227.2 +/- 224.5 nmol/l and 302.1 +/- 212.7 nmol/l in the patients who recovered (p > 0.05). Adrenocortical insufficiency was detected in 16.3% of the patients and 42.9% of these patients died. In conclusion, sepsis is characterized by high basal cortisol level which may show a poor prognosis and a blunted cortisol response to ACTH stimulation. A small percentage of patients with sepsis may develop adrenocortical insufficiency. PMID- 9228517 TI - Alternative strategies in muscle genotype and phenotype studies. A model of intrafusal muscle fibre type differentiation. AB - The development and regeneration of muscle fibres start from myoblasts of embryos or adult animals. The resulting phenotype is a combination of genetically fixed properties of myoblast cell lineages and of extrinsic, primarily neurogenic factors. Intrafusal fibre types of muscle spindles differ from each other and from extrafusal fibres by their ultrastructure, by the presence of both sensory and motor innervation, and by the content of specific myosin heavy chain (MHC) isoforms. Differentiation of these distinctions depends on the morphogenetic influence of primary afferent neurones. It is, however, not known, whether the intrafusal phenotype can be induced in any myotube regardless of its cell line origin or only in a special predetermined intrafusal lineage(s) committed to differentiate into intrafusal muscle fibres. The aim of our studies was to define the contribution of intrinsic myogenic properties of muscle cell lineage and extrinsic neurogenic factors by the sensory and the motor innervation on the differentiation of intrafusal phenotypes using ultrastructural analysis and immunocytochemical determination of MHCs under various experimental conditions. The presented minireview is based on the results of our previous findings, and preliminary experiments indicate that new important results may be obtained in studies of myogenesis and muscle regeneration. PMID- 9228518 TI - Changes in cardiac contractility in IDDM and NIDDM diabetic rats. AB - Differences in myocardial contractility were studied in type I (insulin dependent, IDDM) and type II (non-insulin-dependent, NIDDM) diabetic rats. Using the streptozotocin-induced diabetes as an experimental model, the contractile properties of left ventricular myocardium of IDDM and NIDDM animals were compared to similar parameters of their age-matched controls. Contraction force was analyzed as a function of the pacing frequency. Paired-pulse stimulation and catecholamine treatment were applied to compare the inotropic responses obtained in the two types of diabetes. Diabetic and control preparations developed equal peak tension at each driving frequency upon the application of paired-pulse stimulation with fixed interpulse interval. The interpulse interval dependence of paired-pulse induced inotropy was altered and the velocity of contraction and relaxation decreased in IDDM, but not in NIDDM muscles. Sensitivity to isoproterenol and norepinephrine was decreased in both types of diabetes, however, the isoproterenol resistance of old diabetic animals was attributable to age rather than to the diabetic state. The results indicate that alterations in the contractile parameters and catecholamine sensitivity in IDDM differ from those observed in NIDDM form of diabetes mellitus. PMID- 9228519 TI - The ionic basis of membrane potential changes from before fertilization through the first cleavage in the egg of the frog Rana cameranoi. AB - Experiments were performed to identify the ionic basis of membrane potential changes in the Rana cameranoi egg, from prior to fertilization through the first cleavage. The membrane potential was monitored continuously during this period. Ten per cent Ringer was used as the recording solution in the control group. The effects of Na+ or Ca2+ conductances were observed by altering external concentrations of these ions. K+ and Cl- conductances were tested by adding channel blockers of these ions (TEA and SITS, respectively) to the extracellular medium. The resting potential of the unfertilized egg is mainly affected by K+ conductance. Chloride conductance is responsible for the depolarization phase of the fertilization potential evoked by sperm entry, and K+ conductance is responsible for the repolarization phase of this potential. We suggest that Na+ permeability does not directly contribute to the fertilization potential; however fertilization potential peak is significantly reduced upon a reduction of extracellular sodium. The fertilization potential is not significantly influenced by extracellular Ca2+, and eggs fertilized in calcium-free solutions maintain their normal development; these results suggest that extracellular Ca2+ does not significantly contribute to the electrical and mechanical blocks that prevent polyspermy. The membrane potential of the fertilized egg does not alter significantly until the first cleavage. Potassium conductance contributes to hyperpolarization generated upon the first cleavage, whereas sodium is the basic ion responsible for the phase which follows peak hyperpolarization, and which plays a role in the return of the post-cleavage membrane potential to a steady level. Cl- conductance, which is important as the ionic basis of the fertilization potential, does not significantly influence any parameter of the cleavage cycle. PMID- 9228520 TI - Brownian dynamics simulation of pH-effects on conductance through potassium channels. AB - A new theory termed "tunnel-acid-group-potential" (TAGPT), explaining the effect of pHo and pH(i) on ion conductance through different membrane channels, is presented. It is suggested that shifts in pHo and pH(i) lead to changes in values of negative charges generated by acid groups of side chains of some polar (Glu-, Asp-) amino acid residues, lining the tunnel part of the channel. The resulting modification of electrostatic field influences the heights of rate-limiting energy barriers (for ion transport) in the transition zones between the tunnel and the vestibules, followed by changes in channel conductivity. PMID- 9228521 TI - Analysis of kinetic properties of gamma-glutamyl transpeptidase from rat kidney. AB - The initial rate kinetics of rat kidney gamma-glutamyl transpeptidase were measured using L-gamma-glutamyl-p-nitroanilide and glycyl-glycine as the donor and the acceptor substrate, respectively. Experimental data were fitted with the initial rate equation, and the obtained results indicated that: (1) Michaelis constants for transpeptidation (Kb), autotranspeptidation (Ka), and hydrolysis (Kh) are 8.56 mmol/l, 2.02 mmol/l and 0.005 mmol/l, respectively. (2) The maximum rate of transpeptidation (Vb) exceeds that of hydrolysis (Vh) and autotranspeptidation (Va) 160 times and 5 times, respectively. (3) A comparison of the ratios of maximal rate: Michaelis constant of individual reactions shows that hydrolysis is approximately 10 times more efficient than the remaining two reactions. (4) Under routine conditions used for gamma-glutamyl transpeptidase estimation, transpeptidation is the prevalent reaction. PMID- 9228522 TI - The influence of external surface potential and transmembrane potential on the passive transbilayer movement of phospholipids in the red blood cell membrane. AB - The passive transbilayer movement of spin-labelled analogues of phosphatidyl choline (PC), phosphatidyl-ethanolamine (PE), and phosphatidyl-serine (PS) in red blood cell membranes was investigated at physiological and low ionic strength of the extracellular solution. Passive transbilayer movement of aminophospholipids PS and PE was measured in ATP-depleted cells. To discriminate between a possible surface potential and a transmembrane potential effect, NaCl in physiological ionic strength solution was replaced either by sucrose or by Na-tartrate (constant osmolarity). Neither in sucrose (low ionic strength) nor in Na-tartrate media a significant change of the translocation rate of the phospholipids was observed. From these results in can be concluded that changes of the external surface potential as well as of the transmembrane potential do not affect the passive transbilayer movement of phospholipids in human red blood cells. PMID- 9228523 TI - Human hippocampus establishes associations in memory. AB - Studies of amnesia have demonstrated that the hippocampus is necessary for long term memory, but its precise role in memory is unknown. We designed a positron emission tomography experiment with tailored encoding and retrieval tasks that permitted the isolation of different mnemonic functions theorized to be mediated by the hippocampus. These functions included encoding single items, establishing interitem associations, novelty detection, and retrieving recently formed associations. Of these, we found hippocampal and parahippocampal activation only during associative learning. Our results indicate that the hippocampal formation may be particularly involved in the establishment of associations among components of an episode in memory. PMID- 9228524 TI - Piriform cortex efferents to the entorhinal cortex in vivo: kindling-induced potentiation and the enhancement of long-term potentiation by low-frequency piriform cortex or medial septal stimulation. AB - The entorhinal cortex receives input from many cortical areas and mediates the flow of information between these sites and the hippocampal formation. Long-term synaptic plasticity in cortical efferents to the entorhinal cortex may contribute to the transmission of neural activity to the hippocampus, as well as the storage of information, but little is known about plasticity in these pathways. We describe here the use of evoked field potential recordings from chronically implanted electrodes in the rat entorhinal cortex to investigate synaptic plasticity in the large piriform (olfactory) cortex projection to the superficial layers of the entorhinal cortex. Both kindling-induced potentiation and long-term potentiation (LTP) were tested. In addition, we attempted to modulate LTP induction by the co-induction of frequency potentiation and by the co-activation of the medial septum. Epileptogenic kindling stimulations of the piriform cortex (1-s, 60-Hz trains 3 times/day for 5 days) were found to result in a reliable potentiation of field responses evoked by piriform cortex test pulses. Non epileptogenic tetanization of the piriform cortex with 400-Hz 16-pulse trains reliably resulted in LTP effects. These effects could be augmented by embedding brief LTP induction stimuli within 11-pulse, 15-Hz trains that alone produce only frequency potentiation. Co-activating the medial septum with 10-Hz trains, just prior to tetanization of the piriform cortex, augmented LTP of piriform cortex inputs to the entorhinal cortex in an input-specific manner. All potentiation effects were found to last for periods of weeks. These findings demonstrate that both epileptogenic and non-epileptogenic piriform cortex stimulation induces lasting potentiation of population field responses in the entorhinal cortex of the awake rat. The LTP effects were inducible in a graded manner and were sensitive to the temporal context of stimulation. The finding that low-frequency activation of the septum can enhance plasticity in the entorhinal cortex adds to a body of data indicating a role for the medial septum in contributing to theta activity and plasticity in both the entorhinal cortex and hippocampal formation. PMID- 9228525 TI - gamma Isoform-selective changes in PKC immunoreactivity after trace eyeblink conditioning in the rabbit hippocampus. AB - An immunocytochemical examination of the rabbit hippocampus was done to determine which of the Ca(2+)-dependent protein kinase C (PKC) isoforms (PKC alpha, -beta I, -beta II, or -gamma) are involved in associative learning. The hippocampally dependent trace eyeblink conditioning task was used for behavioral training, and pseudoconditioned and naive animals served as controls. Significant increases (P < 0.05) in staining intensity were found with antibodies reactive with the catalytic or the regulatory domain of PKC gamma in conditioned animals compared with naive and pseudoconditioned subjects at a 24-h post-conditioning time point. The increase was found in CA1 and CA3 pyramidal cell bodies, in apical dendrites and the proximal part of the basilar dendrites, and in cell bodies of dentate granule cells. In contrast, no conditioning-specific changes were found for PKC alpha, -beta I, or -beta II in in hippocampal neurons. The increase in PKC gamma immunoreactivity (ir) was significantly less (P < 0.05) in poor learners than in good learners. The correlation between the degree of PKC gamma-ir and the total number of conditioned responses across training sessions was both positive and significant. These results suggest that PKC gamma is the major Ca(2+)-dependent PKC isoform involved in hippocampal neurons during acquisition of associative memories. Immunoblots revealed no conditioning-induced increase in the total amount or translocation of PKC gamma at the 24-h time point, and no proteolytic PKC fragments were observed. In agreement with the Western blot data, PKC activity did not differ among naive, pseudoconditioned, and trace conditioned animals. The conditioning-induced increase in antibody binding to the gamma isoform must therefore be due to an increased access to the antigenic site(s) as a result of alteration in the tertiary structure of PKC gamma or in quaternary interactions of PKC gamma in situ. PMID- 9228526 TI - Nitric-oxide-guanylyl-cyclase-dependent and -independent components of multiple forms of long-term synaptic depression. AB - Long-term depression (LTD) of synaptic strength is induced by glutamate-triggered increases in postsynaptic [Ca2+], through either influx or release from intracellular stores. Induction of LTD has also been reported to require release of Ca2+ from presynaptic stores and activation of presynaptic Ca2+/calmodulin dependent protein kinase II. This finding leads to the hypothesis that the intercellular messenger nitric oxide (NO) may be a means by which postsynaptic Ca2+ triggers changes expressing LTD in presynaptic terminals. We report that bath application of the oxadiazoloquinoxalone derivative ODQ (4 microM), a selective inhibitor of NO-sensitive guanylyl cyclase (NOGC), markedly attenuated (90%) the magnitude of LTD induced by low-frequency stimulation (LFS; 1 Hz/15 min) of Schaffer collateral-CA1 synapses in hippocampal slices in vitro. Both the NO donor S-nitroso-N-acetylpenicillamine (100 microM) and the membrane-permeant cyclic guanine 3',5'-monophosphate (cGMP) analogue 8-(-4-chlorophenylthio) guanosine (8-pCPT)-cGMP (50 microM) enhanced the magnitude of LTD, which is consistent with he hypothesis that activation of NOGC plays a role in the induction of LTD. Nicotinamide (20 mM), an inhibitor of NO-activated ADP ribosyltransferase, did not impair the induction of LTD. In contrast to de novo LTD, the reversal of long-term potentiation by LFS (depotentiation) was only partially blocked (55%) by ODQ, and heterosynaptic LTD was not impaired at all, suggesting that there are both NOGC-dependent and -independent forms of LTD. Because postsynaptic intracellular infusion of ODQ (500 microM) failed to block the induction of LTD, we conclude that activation of presynaptic NOGC is a necessary step in the induction of an NOGC-dependent component of LTD. PMID- 9228527 TI - Parallel involvement of perirhinal and lateral entorhinal cortex in the polysynaptic activation of hippocampus by olfactory inputs. AB - It has previously been shown that olfactory input to the hippocampus (HPC) is mediated polysynaptically via the lateral entorhinal cortex (LEC), the site of origin of the lateral perforant pathway (LPP). Because previous anatomical studies have shown that olfactory projections also terminate in perirhinal cortex and that this latter region projects directly to the hippocampus, we investigated the role of perirhinal cortex (PRC) in the mediation of the olfactory-hippocampal potential in the rat. Single-pulse stimulation of the lateral olfactory tract (LOT) resulted in a long onset latency (12-20 ms) evoked response in the dentate gyrus of the ipsilateral hippocampal formation. LOT-HPC potentials were rapidly and completely abolished following the microinfusion of procaine into the LPP, suggesting that they are ultimately mediated via this pathway. In support of this finding, current source density analysis indicated that the LOT-HPC response was generated by a current sink at the outer molecular layer of both dorsal and ventral blades of the dentate gurus. Electrolytic and ibotenic acid lesions of PRC produced a significant decrease in the amplitude of LOT-HPC potentials when testing was conducted 4-7 days postlesion. Lesions of LEC produced similar effects and combined lesions of LEC and PRC resulted in an almost complete eradication of the potential, suggesting that parallel entorhinal-hippocampal and perirhinal-hippocampal pathways are involved. These data suggest, therefore, that a portion of the olfactory input to the hippocampus is mediated via polysynaptic connections routed through perirhinal cortex. Because recent research has suggested that PRC plays an important role within the temporal lobe memory system, this connectivity may be important for olfactory memory processes. PMID- 9228529 TI - Postnatal development of zinc-containing cells and neuropil in the hippocampal region of the mouse. AB - The present study describes the postnatal development of zinc-containing boutons and their neurons of origin in the hippocampal region of the mouse. Ages investigated for the development of zinc-containing neuropil were postnatal days 0 (P0), P3, P7, P11, P15, P21, and P28. For zinc-containing cell bodies P7, P15, P21, and P28 were studied. In the area dentata, zinc-containing neuropil appeared first by P3 adjacent to the suprapyramidal limb of the granule cell layer and extended later toward the infrapyramidal limb. By P15, inter- and intralaminar gradients corresponded to those seen in adult animals. The appearance of labeled granule cells followed closely, although temporally delayed, the pattern of granule cell neurogenesis. All granule cells were labeled by P28. In the hippocampus proper, zinc-containing neuropil was seen by P0, but staining of the incipient mossy fiber zone was first visible by P3. Staining pattern and intensity developed gradually until they reached their mature appearance by P15. The distribution of labeled cells was identical to that seen in mature animals by P7 in CA3, but first by P21 in CA1. In the subiculum, neuropil staining first appeared proximally by P7, included all of this area by P11, and appeared mature by P21. A few labeled cells were seen in the proximal subiculum at all ages at which labeled cells were present in CA1. Labeled cells which extended further distally became first visible by P21. Their number and labeling intensity reached mature levels by P28. In the presubiculum, retrosplenial area 29e, and parasubiculum, neuropil staining first appeared by P3. The retrosplenial area 29e could be distinguished by P11. This area and the presubiculum reached their adult appearance by P21. This occurred first by P28 in the parasubiculum due to the late maturation of the parasubiculum a. Labeled cells were first seen by P7 in layer III of the presubiculum and by P15 in the retrosplenial area 29e and the parasubiculum. Cell labeling appeared mature by the same times as the neuropil staining. In the entorhinal areas a very light neuropil stain was apparent in the deeper layers by P0. A distinct rise in staining intensity was first observed by P7 in layers I-III. Thereafter, mature characteristics developed gradually and were attained by P21. Cell labeling was not seen in the medial entorhinal area. A few labeled cells were apparent by P7 in the lateral entorhinal area. After a slight increase by P15, numerous labeled cells were found in layer II and layer VI by P21. Their distribution and labeling intensity appeared mature by P28. Zinc containing cells appear to represent cells formed late in the course of neurogenesis in all areas aside from the lateral entorhinal area. As far as intrinsic connections are concerned, it is the development of projections from this subset of neurons which is monitored in this study. We suggest that the appearance of zinc may contribute via its different effects on N-methyl-D aspartate (NMDA) and non-NMDA glutamate receptors to the end of a developmental phase that is permissive to changes in synaptic efficacy. Species differences and alternative functions of zinc are considered. PMID- 9228528 TI - Distribution, ultrastructure, and connectivity of calretinin-immunoreactive mossy cells of the mouse dentate gyrus. AB - Hilar mossy cells of the mouse were shown recently to display calretinin immunoreactivity (Liu et al. [1996] Exp Brain Res 108:389-403). The morphological and connectional characteristics of these cells are poorly understood. In the present study, we used immunohistochemical, electron microscopic, and neuronal tracing techniques to describe their distribution, morphology, and connectivity. The distribution of calretinin-immunoreactive mossy cells varied significantly along the dorsoventral axis of the hilus. At dorsal levels, calretinin immunoreactivity was limited largely to a subpopulation of interneurons. At mid dorsoventral and ventral levels, however, most if not all mossy cells displayed calretinin immunoreactivity. We found that most hilar mossy cells are calretinin immunoreactive but lack gamma-aminobutyric acid, as demonstrated by postembedding immunostaining of alternate semithin sections. Calretinin-immunoreactive mossy cells typically had two to three thick dendrites covered with complex spines (thorny excrescences). Electron microscopy revealed that these spines received multiple asymmetric contacts from mossy fibres. Axons arising from these cells formed a strong belt of calretinin immunoreactivity restricted to the inner third of the dentate molecular layer. This immunoreactivity was equally dense throughout the dorsoventral length of the dentate gyrus, suggesting that axons of calretinin-immunoreactive mossy cells located in the ventral levels diverge greatly and are capable of innervating distant regions of the dentate gyrus. Ultrastructural examination showed that calretinin-immunoreactive boutons made asymmetric synaptic contacts primarily on spines and, occasionally, on dendritic shafts of granule cells and accounted for the majority of asymmetrical synapses in the inner molecular layer. Injections of the retrograde tracer wheatgerm agglutinin-gold into the dentate gyrus demonstrated that calretinin immunoreactive mossy cells concentrated in the ventral hilus project massively to both the dorsal and ventral aspect of the contralateral dentate gyrus. A small proportion of retrogradely labelled cells showed immunoreactivity for neuropeptide Y or somatostatin. If mossy cells of the ventral hilus receive the majority of their input from ventral granule cells, one may expect ventral granule cells to be more efficient in recruiting large numbers of granule cells during synchronous activity patterns than dorsal granule cells. Spontaneous activity originating from granule cells in the ventral dentate gyrus can be propagated throughout the dorsoventral length of the dentate gyrus bilaterally via the dorsoventrally divergent and contralaterally projecting axons of the mossy cells. This organization may explain why the ventral dentate gyrus is frequently involved in pathological phenomena. PMID- 9228531 TI - Origin of the lateral perforant path. PMID- 9228530 TI - Adhesion molecule expression on phagocytic microglial cells following anterograde degeneration of perforant path axons. AB - Entorhinal cortex lesion (ECL) leads to anterograde degeneration of perforant path axons and is known to induce a rapid and intense reaction of astrocytes and microglial cells in the deafferented dentate gyrus. Phagocytosis of degenerating axons involves the establishment and maintenance of cell-matrix and cell-cell interactions by activated glial cells. It was thus our aim to investigate whether the process of axon phagocytosis is accompanied by the expression of adhesion molecules on activated microglial cells or reactive astrocytes, as such molecules mediate bot cell-matrix and cell-cell interactions. We found that the integrin adhesion molecules leukocyte function antigen-1 (LFA-1), very late antigen-4 (VLA 4), and the ligand for LFA-1, intercellular adhesion molecule-1 (ICAM-1), were expressed on microglial cells accumulating in the outer molecular layer of the deafferented dentate gyrus. This upregulation of adhesion molecule expression on microglial cells showing morphological criteria of activation occurred rapidly following ECL, reached its peak at 3 days post lesion (dpl), and gradually returned to control levels after 9 dpl. Astrocytes were never labeled by antibodies directed against these adhesion molecules. Prelabeling of the perforant path with a fluorescent tracer and subsequent ECL led to phagocytosis of fluorescent-labeled axonal debris by cells that were located in the outer molecular layer and showed typical microglial morphology. Double-fluorescence labeling demonstrated that microglial cells engaged in the phagocytosis of axonal debris expressed LFA-1, VLA-4, and the LFA-1-ligand ICAM-1. In conclusion, our results demonstrate that anterograde degeneration of perforant path axons results in adhesion molecule expression on activated microglial cells engaged in axon phagocytosis. The expression of such molecules could represent a mechanism that retains activated microglia in areas of axonal degeneration and perhaps enables the interaction of microglial cells with each other or with other immunocompetent cells. PMID- 9228532 TI - Award ceremony of the E.K. Frey-E. Werle Foundation of the Henning L. Voigt family. PMID- 9228533 TI - Search for autoantibodies to the human bradykinin B2 receptor. PMID- 9228534 TI - Inhibition of B2 receptor internalization delays its dephosphorylation. PMID- 9228535 TI - Immunolocalization of bradykinin receptors on human synovial tissue. AB - Kinins have been implicated in the pathogenesis of experimental and clinical inflammatory arthritis. Previous studies have reported increased amounts of plasma and tissue kallikreins in synovial fluid, raised kinin levels and an upregulation of kinin B2 receptors on synovial fluid neutrophils in rheumatoid arthritis. Bradykinin binding sites have been identified on human synovial cells by autoradiographic localization and Scatchard analysis. This study was undertaken to localize immunohistochemically kinin B1 and B2 receptors on human synovial tissue. Synovial tissue was obtained at the time of joint replacement surgery or arthroscopic synovectomy in six patients (two RA, two OA and two with avascular necrosis). Tissue sections were immunolabelled for kinin B1 and B2 receptors and viewed by light and confocal microscopy. No immunolabelling of the kinin receptors was observed in the method controls. In all patients labelling for kinin B2 receptors was observed in the synovial lining cells, fibroblasts and endothelial lining cells of blood vessels. There was no immunolabelling for kinin B1 receptors in all samples. These findings further support a role for the B2 receptors in joint diseases. There did not appear to be an induction of the kinin B1 receptor in human synovial tissue obtained from patients with chronic arthritis. However, further studies are required to assess the role of B1 receptors in active joint inflammation. PMID- 9228536 TI - Localization of the bradykinin B2 receptor in uterus, bladder and Madin-Darby canine kidney cells. AB - Kinins are biologically active peptides that act through specific receptors, B1 and B2. Here we describe the localization of the bradykinin B2 receptor in Madin Darby canine kidney cells and in the uterus and urinary bladder of rat or human origin. We discuss the suitability of anti-peptide antibodies to assess the tissue distribution of bradykinin B2 receptors. PMID- 9228538 TI - B1 and B2 kinin receptors in various species. AB - The characteristic features of kinin B1 and B2 receptors are analyzed. Emphasis is placed on the pharmacologic profiles of B1 and B2 functional sites by the use of naturally-occurring kinins, some selective agonists, as well as peptide and non-peptide antagonists. Species differences are indicated, particularly between human, rabbit and mouse B1 receptors and also between human, rabbit and guinea pig B2 receptors. Extensive use has been made of the non-peptide B2 receptor antagonist, WIN-64338 (which is active in the guinea pig and inactive in the other species) and FR-173657 which shows high affinity in human, rabbit, mouse, pig and guinea pig B2 receptors. PMID- 9228537 TI - Evidence for bradykinin B2-receptors on cultured human decidua cells. AB - Bradykinin is known to be present at sites of acute inflammation and to exert its potent inflammatory effects mainly via the bradykinin B2-receptor. Recently, bradykinin dependent processes have been described in cultured human decidual cells, so that bradykinin may expand the list of paracrine factors involved in labour induction. In this paper we present the results of in vitro studies giving evidence that these cells carry the bradykinin B2-receptor. By immunocytochemical methods the receptor protein was localized on decidual cells. Analysis of cellular extracts of cultured decidual cells by RT-PCR showed the presence of the specific mRNA coding for the bradykinin B2-receptor. Binding studies revealed a single, saturable and specific binding site for bradykinin of high affinity (Kd = 0.85 nM, Bmax = 436 fmol/mg protein). Competitive binding studies showed displacement of [3H]-bradykinin by HOE 140, but not by the ligands for the bradykinin B1-receptor, des-Arg10-kallidin and [Leu8]-des-Arg9-bradykinin. The results are consistent with the presence of the bradykinin B2-receptors. PMID- 9228539 TI - Expression of bradykinin B1 receptors in colonic epithelia. AB - The rat colonic epithelium shows no response to [des-Arg9] bradykinin (DAB) in newly isolated tissues, but does so after 3 h in vitro. The EC50 for DAB is circa 50 nM and the maximal response is 57.8 +/- 5.3 microA cm-2 (n = 68). The whole of the current increase is sensitive to frusemide and is predominantly due to electrogenic chloride secretion. The development of responsiveness is prevented by inhibitors of translation (cycloheximide) and transcription (actinomycin D). Thus the receptor for DAB has the hall marks of a classical B1 kinin receptor. While the classical B2 kinin receptor antagonist HOE140, has no effect on the responses, the classical B1 kinin receptor antagonist, [Leu8]DAB, behaves as an agonist in this system. It is concluded that there may be more than one type of B1 kinin receptor. PMID- 9228540 TI - Kinin receptors in human vascular tissue: their role in atheromatous disease. AB - Using samples of many human blood vessels, obtained at autopsy and specific antibodies directed to peptide sequences of the kinin B1 and B2 receptors, we demonstrate the localisation of these receptors within the human vascular system using standard immunolabelling techniques. In large elastic arteries and veins, kinin receptors are present only in the endothelial cells whereas in all muscular arteries and arterioles, these receptors are present in both the endothelial and smooth muscle cells. The identification of kinin receptors in human blood vessels confirms that kinins may modulate both vascular permeability and contractility. The incidental finding at histology, of patchy atheromatous disease in the coronary, femoral, vertebral and pericallosal arteries, assisted in elucidating the role of these receptors in the commonest disease affecting human blood vessels. Intense labelling for B1 receptors was observed in the endothelial cells, foamy macrophages, inflammatory cells and fibroblasts within the thickened intima of the plaque as well as in smooth muscle cells of the underlying tunica media. Immunoreactive B2 receptors were also observed in these cells but with reduced intensity. The intense immunolabelling of B1 receptors in these regions suggest that these may be induced by atheromatous disease and may have therapeutic importance for the B1 receptor antagonists. PMID- 9228541 TI - Kinin receptor status in normal and inflammed gastric mucosa. AB - No documented studies have been reported on the presence of B1 and B2 kinin receptors in the mammalian gastric mucosa. This first study aimed to immunolocalise sites of B1 and B2 kinin receptors in the human pyloric gastric mucosa and to evaluate its role in gastritis. Biopsies were obtained from patients with dyspepsia during endoscopic examination of the patient. The diagnosis and grading of the gastritis was performed on histological examination. Sections were immunostained for both B1 and B2 receptors using rabbit anti-human B1 and B2 kinin receptor antibodies. Control tissue was obtained from partial gastrectomy specimens, following surgical excision of the antrum for duodenal ulcers. The control antrum tissue showed strong immunoreactivity for kinin B2 receptors with positivity noted along the luminal border, at the base of the mucous and stem cells. The B1 receptor was not immunolocalised. Biopsies of all five patients with gastritis showed a decrease in immunolabelling of the B2 receptor and an induction of the B1 receptor especially in regenerating cells. In gastritis there is destruction of the normal mucosal glandular architecture with subsequent regeneration of the epithelial cells. The pyloric glands are infiltrated by acute inflammatory cells that cause crypt abscesses with loss of the epithelial cell membranes. This may explain the reduction in the immunolocalisation of the B2 kinin receptors and the induction of the B1 receptors in active gastritis. Follow up studies after treatment of the inflammation with a combination of B1/B2 kinin receptor antagonists are indicated. PMID- 9228542 TI - Potent, long-acting, orally-active bradykinin antagonists for a wide range of applications. AB - Actions of bradykinin (Arg-Pro-Pro-Gly-Phe-Ser-Pro-Phe-Arg; BK) are mediated by constitutively expressed B2 receptors, that require the full BK peptide chain, and by B1 receptors, induced in inflammation, that use BK(1-8) as ligand. In addition to many physiological and pathophysiological functions, the growth factor activity of BK evidently allows it to act as an autocrine stimulant for small cell lung cancer. A new group of BK antagonists containing the novel amino acid a-(2-indanyl)glycine provides extremely potent broad-spectrum as well as selective antagonists for all these functions. PMID- 9228543 TI - Tachyphylaxis of the B1 kinin receptor in porcine endotoxin shock. AB - Previous experiments in anesthetized pigs have demonstrated that blockade of the bradykinin B2 receptor in experimental endotoxin shock attenuates LPS-induced organ failure, lung dysfunction and mortality. Additional B1 receptor blockade in this situation seems to counteract the beneficial effects of B2 blockade. This suggests that the upregulation of B1 receptors during porcine LPS shock may be a useful mechanism of host defense. Furthermore, infusion of a B1 agonist during septic shock may be of therapeutic benefit. In order to prepare an experiment with B1 stimulation in LPS shock, we conducted a study in anesthetized pigs, in which the B1 receptor has been upregulated by infusion of bacterial lipopolysaccharide (LPS), by evaluating the effect of constant intravenous infusions of the B1 agonist des-Arg10-kallidin on the hypotensive response to bolus doses of this agonist. Following infusions of lipopolysaccharide from S. abortus equi, anesthetised pigs received repeated intra-arterial bolus injections of des-Arg10-kallidin before and during continuous infusions of this agonist in doses of 3, 10, 30 and 100 ng/kg/min. We found that all doses greater than 3 ng/kg/min produced attenuation of the hypotensive response produced by bolus administration of the B1 agonist des-Arg10-kallidin. We conclude that tachyphylaxis is an important feature to be considered in experiments with continuous administration of a B1 agonist in LPS shock. PMID- 9228544 TI - Effect of bradykinin B2-receptor antagonism on post-ischemic coronary reperfusion. AB - Isolated rabbit hearts in a modified Langendorff preparation were used to study the role of bradykinin B2-receptor antagonism on recovery from cardiac ischemia. The experiments were performed to clarify a potential risk of administrating BK receptor antagonists in patients with coronary heart disease and patients undergoing heart surgery. The hearts were perfused in an open system at a constant pressure of 70 mm Hg and at a constant temperature of 37 degrees C with Krebs-Henseleit-buffer solution containing 6.5% HAES (hydroxyethyl-amylopectin) and 10 mmol Na-pyruvate/l. The perfusate was equilibrated to a PO2 of about 500 600 mm Hg. After a steady state period of 30 min, coronary perfusion was stopped for 30 min and the hearts were subsequently reperfused for further 30 min. Pretreatment with the B2-receptor antagonist, CP-0127, significantly increased (p < 0.001) the coronary vascular resistance during reperfusion from 31.9 +/- 2.1 to 59.0 +/- 6.5 mm Hg/ml/min/10 g wt and decreased the left ventricular pressure amplitude to 44.0 +/- 15.2% (p < 0.005) of it's baseline. When ischemia was combined with hyperkalaemic cardioplegia, CP-0127 did not influence coronary vascular resistance, but depressed, only to a minor degree, left ventricular pressure amplitude to 65.1 +/- 15.4% (p < 0.005) during reperfusion. Arrhythmias during reperfusion with cardiac arrest occurred only in 2 hearts with B2-receptor inhibition when ischemia was not combined with cardioplegia. Dependent on the initial functional status of the heart B2-receptor inhibition impaired recovery from acute coronary ischemia by increasing the coronary vascular resistance and depressing the myocardial function and therefore may constitute a risk in patients undergoing heart surgery and extracorporeal circulation. PMID- 9228545 TI - Possible protective effects of kinins and converting enzyme inhibitors in cardiovascular tissues. AB - The main objective of this study was to determine if the components of the kallikrein-kinin system are released into the venous effluent from isolated perfused rat hearts. To assess the contribution of kinins and the vascular and cardioprotective effects of the ACE inhibitor ramipril, we determined the status of cardiac kallikrein (CKK), potent kinin-generating enzyme, in rats with right ventricular hypertrophy induced by chronic volume overload and left ventricular hypertrophy by aortic banding. CKK was measured as previously described (Nolly, H.L., Carbini, L., Carretero, O.A., Scicli, A.G., 1994). Kininogen by a modification of the technique of Dinitz and Carvalho (1963) and kinins were extracted with a Sep-Pak C18 cartridge and measured by RIA. CKK (169 +/- 9 pg Bk/30 min), kininogen (670 +/- 45 pg Bk/30 min) and immunoreactive kinins (62 +/- 10 pg Bk/30 min) were released into the perfusate. The release was almost constant over a 120 min period. Pretreatment with the protein synthesis inhibitor puromycin (10 mg i.p.) lowered the release of kallikrein (42 +/- 12 pg Bk/30 min, p < 0.001) and kininogen (128 +/- 56 pg Bk/30 min, p < 0.001). Addition of ramiprilat (10 micrograms/ml) increased kinin release from 54 +/- 18 to 204 +/- 76 pg Bk/30 min (p < 0.001). Aortic banding of rats increased their blood pressure (BP) (p < 0.001), relative heart weight (RHW) (p < 0.001) and CKK (p < 0.001). Ramipril treatment induced a reduction in BP (p < 0.05) and RHW (p < 0.005) while CKK remained elevated. Aortocaval shunts increased their ANF plasma levels (p < 0.05), RHW (p < 0.001) and CKK (p < 0.01). Ramipril treatment induced a reduction in RHW (p < 0.05), while CKK and ANF increased significantly (p < 0.05). The present data show that the components of the kallikrein-kinin system are continuously formed in the isolated rat heart and that ramipril reduces bradykinin breakdown with subsequent increase in bradykinin outflow. The experiments with aorta caval shunt and aortic banding show that cardiac tissues increase their kinin-generating activity and this was even higher in ramipril treated animals. This may suggest that the actual level of kinins is finely tuned to the local metabolic demands. In this experimental model of cardiac hypertrophy. ACE inhibitors potentiate the actions of kinins and probably try to normalise endothelial cell function. PMID- 9228546 TI - Physical and biological significance of peptide sequences mediating the interaction between high molecular weight kininogen and plasma prekallikrein. AB - HK31 (S565-K595) has previously been shown to encompass the binding domain for plasma prekallikrein (PK) within domain 6 of high molecular weight kininogen (HK). The complementary binding domain for HK within PK is mapped to PK56 (F56 G86), in the Apple 1 domain and to PK266 (K266-C295) in the Apple 4 domain. Isothermal titration calorimetry demonstrated that either PK peptide binds to HK31 in 1:1 stoichiometry. Binding of the alternate PK peptide into a ternary complex is facilitated nearly 2-fold. Fluorescence emission spectroscopy revealed that only the binding of PK56 caused a limited decrease in intrinsic tryptophane fluorescence emission intensity of HK31. We conclude that the two PK peptides bind to the HK peptide at different sites. To map the minimal sequence within HK31, truncated new peptides were tested for their ability to compete with HK for binding PK in a cell-free system. D567-T591, a 25-residue peptide which contains sufficient structural information for binding kallikrein in solution, blocked the binding of kallikrein to HK bound to endothelial cells and inhibited PK activation to kallikrein and the generation of kallikrein-activated urokinase on endothelial cell surfaces. HK-derived peptides could modulate excessive fibrinolysis and hypotension in sepsis and multiple trauma. PMID- 9228548 TI - Generation of kinins in synovial fluid from patients with arthropathy. AB - An enzyme-linked immunosorbent assay method is described for the measurement of kinin formation in synovial fluid from patients with rheumatoid and osteoarthritis (RA and OA). Basal kinin concentrations were less than 6 ng/ml in synovial fluid collected in the presence of inhibitors of kinin forming (kininogenase) and kinin metabolising (kininase) enzymes. During incubation of synovial fluid in the presence of kininase inhibitors alone, kinins were produced rapidly over the first 10 min, but production ceased completely within 30 min due to inhibition of the endogenous kininogenases; the rate of kinin generation during the early rapid phase correlated well with the plateau kinin concentration. Plateau kinin levels in synovial fluid from 15 patients with OA and RA ranged from 98 to 427 ng/ml, with a median value of 148 ng/ml. This study demonstrates clearly that synovial fluid from arthritis patients has the capacity to produce kinins. Although the number of patients was small, the amount of kinin generated in vitro varied over a wide range and a relationship between intra articular kinin formation and clinical features may become apparent in a larger group of patients. The technique could also be used to investigate other biological systems in which a role has been proposed for kinins. PMID- 9228547 TI - Binding of activation of kinin-forming proteins on vascular endothelial cells. PMID- 9228549 TI - Tissue kallikrein and the bradykinin B2 receptor are expressed in endometrial and prostate cancers. AB - KLK1/tissue kallikrein is a member of a family of structurally related serine proteases which exhibit a diverse range of enzymic activities. Other members of this family of genes and the bradykinin (B2) receptor have been implicated in cancer processes. The KLK genes are expressed in various organs of the reproductive tract, including the prostate and uterus. In this study, we have examined whether KLK1/tissue kallikrein and the B2 receptor were both expressed in cancers emanating from these organs. We have shown that KLK1/tissue kallikrein and the B2 receptor are expressed, to varying degrees, in prostate disease and 2 human prostate cancer cell-lines, LNCaP and DU145. KLK1 is also expressed in a range of endometrial cancers, although at lower levels than that observed for normal human endometrial tissues. These findings suggest that a functional kallikrein-kinin system is present in prostate disease, in particular, which may be important in the process of tumorigenesis in this tissue. PMID- 9228550 TI - Expression of human tissue kallikrein in rat salivary glands and its secretion into circulation following adenovirus-mediated gene transfer. AB - Replication-deficient adenovirus Ad.CMV-cHK, expressing human tissue kallikrein under the control of the cytomegalovirus enhancer/promoter, was introduced into rat salivary glands via a direct intracapsular injection. A single injection of Ad.CMV-cHK at a dose of 4 x 10(9) pfu resulted in a sustained expression of human tissue kallikrein in rat salivary glands. The level of immunoreactive human tissue kallikrein in rat sera was the highest at 1 day post gene delivery when both salivary glands were injected and decreased in a time-dependent manner after gene delivery. Human tissue kallikrein levels in sera increased concomitantly with the amount of adenovirus used in direct salivary injection. The detection of human tissue kallikrein in sera after gene delivery into salivary glands provided direct evidence indicating that rat salivary glands secrete locally synthesized human tissue kallikrein to the systemic circulation. The direct injection of salivary glands with replication-deficient adenovirus could provide a systemic route for gene delivery for studying salivary gland function and development. Targeted gene delivery to the salivary gland may provide the means to express therapeutic proteins in saliva and the systemic circulation. PMID- 9228552 TI - Role of the renal kallikrein-kinin system in the development of hypertension. AB - Role of renal kallikrein-kinin system has been studied using mutant Brown-Norway Katholiek (BN-Ka) rats, in which both high- and low-molecular weight kininogens were almost absent in plasma and kinin in urine was mainly not detectable. Mutant BN-Ka rats were very sensitive to increased salt intake, resulting in raised systemic blood pressure that is linked to reduced urinary excretion of sodium, when compared with normal BN-Kitasato (BN-Ki) rats. Consequently, sodium accumulated in erythrocytes and cerebrospinal fluid in mutant BN-Ka rats. Subcutaneous infusion of angiotensin II (20 mg/day/rat) also enhanced the concentration of sodium in erythrocytes and in cerebrospinal fluid and increased the systemic pressure by releasing aldosterone. A 4-day infusion of 0.3 M sodium solution (6 ml/kg/h) to the abdominal aorta of conscious and un-restrained mutant BN-Ka rats increased the pressor responses of the arterioles to norepinephrine and angiotensin II (i.a.) by 30- and 10-fold, respectively. Infusion of ebelactone B, (a selective inhibitor of carboxypeptidase Y-like exopeptidase, a kininase in rat urine), to normal BN-Ki rats during induction of hypertension with DOCA and salt, resulted in the reduction of the raised blood pressure, indicating that a site of action of kinins was at the luminal membrane of the renal tubule cells. Our results support the view that the role of renal kallikrein-kinin system is to excrete 'excess sodium' and a reduction in the generation of renal kinins may be a factor in the development of hypertension as a result of the sodium accumulation in the body. PMID- 9228551 TI - Kallikrein gene therapy: a new strategy for hypertensive diseases. AB - The tissue kallikrein-kinin system has been postulated to play a role in blood pressure homeostasis and the pathogenesis of clinical hypertension. To demonstrate the potential therapeutic effects of somatic gene delivery in treating hypertension, we used spontaneously hypertensive rats (SHR) as a model. The gene encoding the human tissue kallikrein was used because of its powerful hypotensive action. The human kallikrein DNA constructs were placed under the control of the metallothionein metal response element, the cytomegalovirus promoter/enhancer or the Rous sarcoma virus 3'-LTR. The human tissue kallikrein DNA constructs were incorporated into adenoviral vectors via homologous recombination. The naked plasmid DNA constructs or adenovirus containing the kallikrein gene were first introduced into kidney 293 cells and the expression of human tissue kallikrein was identified by ELISA. The kallikrein gene was delivered into SHR via intramuscular, intravenous, portal vein, intraperitoneal, and intracerebroventricular routes. A single injection of naked human kallikrein DNA constructs caused a prolonged reduction of high blood pressure for up to 8 weeks. Adenoviral-mediated gene delivery results in high efficiency of human tissue kallikrein expression. Immunoreactive human kallikrein was detected in rat serum at the highest level at 1 day post gene delivery. Portal vein delivery of a reporter gene, AdCMV-LacZ, results in intense staining of beta-galactosidase in rat liver, suggesting that recombinant kallikrein is mainly produced in liver and secreted into the circulation. These results show that kallikrein gene delivery causes a sustained reduction of blood pressure in genetically hypertensive rats and provide important information for a potential gene therapy approach to human hypertension and related diseases. PMID- 9228553 TI - Changes in urinary tissue kallikrein excretion in black African women with hypertensive disorders of pregnancy. AB - The aim of this study was firstly to establish whether tissue kallikrein (TK) was involved in the aetiology of hypertensive disorders of pregnancy. Secondly, to assess whether tissue kallikrein:creatinine ratios may differentiate normotensive pregnant women from those with hypertensive disorders of pregnancy and have a predictive value. Random untimed urine samples were collected from all women (n = 264) recruited to this study. Urine specimens were analyzed for urinary tissue kallikrein using a selective, synthetic chromogenic tripeptide substrate (H-D-Val Leu-Arg-pNA). Urinary creatinine levels were measured using standard methods. There was a significant difference in the excretion of urinary tissue kallikrein between normotensive pregnant women (2.91 ng TK/microgram protein) and women with mild (2.52 ng TK/microgram protein; p < 0.0001) and severe (1.53 ng TK/microgram protein; p < 0.0001) hypertension in pregnancy. No statistical difference was observed with regard to urinary tissue kallikrein excretion between normotensive pregnant and normotensive non-pregnant women (2.87 ng TK/microgram protein; p = 0.16). A positive correlation was observed between the diastolic blood pressure and urinary tissue kallikrein excretion in women with hypertensive disorders of pregnancy. When compared to the normotensive pregnant group, the urinary kallikrein:creatinine ratios were significantly lower in the mild (0.6 versus 0.3; p < 0.0001) and severe (0.6 versus 0.12; p < 0.0001) hypertensive groups. The urinary creatinine excretion was significantly higher in the mild (9.55 +/- 2.6 mmol/l; p < 0.0001) and in severe (15.62 +/- 5.48 mmol/l; p < 0.0001) hypertensives when compared to normotensive pregnant values (5.65 +/- 2.6 mmol/l). The reduced urinary tissue kallikrein excretion in hypertensive disorders of pregnancy may be a significant factor in the development of the hypertension in pregnancy. Measurement of urinary tissue kallikrein: creatinine ratios may represent a simple and practical predictive test to differentiate women with hypertensive disorders of pregnancy from normotensive pregnant women. PMID- 9228554 TI - Localisation of tissue kallikrein in the kidney of black African women with early onset pre-eclampsia: a pilot study. AB - Increased renal production of vasodilator mediators like kinins would counteract the vasospasm of pre-eclampsia. This study examines the cellular localisation of tissue kallikrein (TK), the potent kinin forming enzyme within the nephron of patients with early onset pre-eclampsia. Using the peroxidase-antiperoxidase immunoenzyme complex, TK was immunolocalised in the principal cells of the distal connecting tubule and the cortical collecting duct cells of the distal nephron of control tissue. Moderate reactivity was observed in the epithelial cells lining the Bowmans capsule. In early onset pre-eclampsia, TK was additionally localised in the proximal tubule cells, however, the intensity of reactivity was reduced when compared to that of the distal tubule cells. In patients with hypertension of pregnancy, the occurrence of TK in the proximal tubule suggests either gene induction or emiocytosis of TK. PMID- 9228555 TI - Tissue kallikrein in transplant kidney. AB - Literature survey, thus far, has shown a decrease in the excretion of urinary tissue kallikrein (TK) in transplant patients with a further reduction of the enzyme during episodes of acute rejection. The study aims were to compare, at cellular and subcellular levels, the localisation of tissue kallikrein in biopsies of the transplant kidney to autopsy derived normal renal tissue. Renal biopsies from eighteen transplant patients with deteriorating renal function were obtained. Immunolabelling for tissue kallikrein, using a polyclonal goat anti-TK, antibody raised against recombinant TK, was performed following routine enzymatic, immunofluorescence and electron microscopic techniques. In normal kidney tissue, TK was immunolocalised in the distal connecting tubules and collecting ducts. By comparison the renal transplant tissue showed a reduction in the intensity of label, but maintained the sites of localisation. In the sections examined by electron microscopy, although TK was confined mainly at the luminal side of the cell, some label was noted along the basolateral membranes. In the transplant kidneys, there was a reduction in the overall number of gold particles counted, which correlated with the decreased intensity observed on immunocytochemistry. In addition, there was a shift to a basolateral orientation of the immunolabel. Acute rejection is characterised by oedema, tubulitis and vasculitis. Destruction of the tubule cells and leakage of TK into the interstitial tissue space and the resultant effect of the formed kinins on renal capillary vasculature could explain the observed renal parenchymal oedema and transplant rejection. PMID- 9228556 TI - The evaluation of tissue kallikrein in Helicobacter pylori-associated gastric ulcer disease. AB - Helicobacter pylori (Hp) associated ulcer disease is a common form of gastric disorder involving mucosal damage and invasion of the mucosa by polymorphic inflammatory cells with concomitant changes in the epithelial cell structure. The bacteria are thought to adhere by specific junction zones to the epithelial cell surface resulting in the degeneration of the mucosal layer. Our study was undertaken to examine the relative status of tissue kallikrein (TK) in antral and fundic biopsies, endoscopically obtained from 10 patients suspected of having gastric disorders. For histological evidence of inflammation the tissue was stained with hematoxylin and eosin and classified as mild, active, chronic and chronic active gastritis. Hp infection was determined by Giemsa staining. For localisation of TK, slide-mounted tissue sections were subjected to PAP and immunofluorescent staining using a goat anti-human TK IgG antibody. The results revealed that in the antral control tissue, removed during partial antractomy, TK was immunovisualised along the luminal border of the deep pyloric glands. The surface epithelia and superficial glands showed no labelling. The fundic control tissue revealed an absence of TK in the superficial and surface epithelial glands, but was positive in the parietal cells. The fundic biopsy specimens showed similar immunoreactivity in these areas. By contrast, in the inflammed pyloric mucosa, there was a shift of TK localisation to the basal part of the glandular cells and there was also expression of TK in the superficial glands that showed histological evidence of regeneration. In the fundic biopsies there was no change observed in the sites of TK localisation (similar to control tissue). It was observed, that even though 8 of the 10 subjects exhibited Hp infection, the inflamed mucosa showed no discernable difference in the staining patterns between the infected and non-infected tissue sections. Our findings suggest an important role for a B1/B2 kinin antagonist in patients with gastritis. PMID- 9228557 TI - Proteolytic release of membrane proteins: studies on a membrane-protein solubilizing activity in CHO cells. AB - Diverse membrane proteins are solubilized by a specific proteolytic cleavage in the stalk sequence adjacent to the membrane anchor, with release of the extracellular domain. Examples are the amyloid precursor protein, membrane-bound growth factors and angiotensin-converting enzyme (ACE). The identities and characteristics of the responsible proteases remain elusive. We have studied this process in Chinese hamster ovary (CHO) cells stably expressing wild-type ACE (WT ACE) or juxtamembrane (stalk) deletion or chimaera mutants. Determination of the C termini (i.e. the cleavage sites) of released, soluble wild-type and mutant ACE by MALDI-TOF mass spectrometry indicated that the membrane-protein-solubilizing protease (MPSP) in CHO cells is not constrained by a particular cleavage site motif or by a specific distance from the membrane, but instead may position itself with respect to the putative proximal, folded extracellular domain adjacent to the stalk. Nevertheless, kinetic analyses of release rates indicated that a minimum distance from the membrane must be preserved. Interestingly, soluble full-length (anchor-plus) WT-ACE incubated with fractions of, or intact, CHO cells was not cleaved. In all cases, release was stimulated by a media change or by the addition of phorbol ester, with rate enhancements of 5- and 50-fold, respectively, for WT-ACE. The phorbol ester effect was abolished by staurosporine, a protein kinase C (PKC) inhibitor. We propose that the CHO cell MPSP that solubilizes ACE: (1) only cleaves proteins embedded in a membrane; (2) requires an accessible stalk and cleaves at a minimum distance from both the membrane and proximal extracellular domain; (3) positions itself primarily with respect to the proximal extracellular domain and (4) is regulated in part by a PKC-dependent mechanism. PMID- 9228558 TI - Protein C inhibitor (PCI) and heparin cofactor II (HCII): possible alternative roles of these heparin-binding serpins outside the hemostatic system. PMID- 9228559 TI - Characterization of a multicatalytic proteinase complex (20S proteasome) from Trypanosoma brucei brucei. AB - African trypanosomes are tsetse-transmitted protozoan parasites that cause sleeping sickness in humans and 'Nagana' in animals. A high relative molecular mass multicatalytic proteinase complex (MCP) was purified and biochemically characterized from the cytosolic fraction of Trypanosoma brucei brucei. The isolation procedure consisted of fractionation of the lysate by high speed centrifugation, chromatography on Q-sepharose molecular sieve filtration on Sephacryl S-300, chromatography on HA-Ultrogel and glycerol density gradient centrifugation (10-40%). The final enzyme preparation yielded a single protein band corresponding to a relative molecular mass of 630 kDa on a non-denaturing polyacrylamide gel. The enzyme hydrolyses a wide range of peptide substrates characteristic of chymotrypsin-like, trypsin-like, peptidylglutamylpeptide hydrolysing activities determined by fluorogenic peptides, Z-Gly-Gly-Leu-NHMec, Z Arg-Arg-NHMec and Z-Leu-Leu-Glu-beta NA, respectively. The enzyme was found to have a wide variation in pH optimal activity profile, with optimum activity against Z-Gly-Gly-Leu-NHMec at 7.8, Z-Arg-Arg-NHMec at pH 10.5 and Z-Leu-Leu-Glu beta NA at pH 8.0, showing that the different activities are distinct. The enzyme hydrolysed oxidized proteins. In addition, the chymotryptic and trypsin-like activities were susceptible to inhibition by peptide aldehyde inhibitors with variable inhibition effects. The study demonstrates the presence of a non lysosomal proteasome pathway of intracellular protein degradation in the bloodstream form of T. b. brucei. Further, the ability of the enzyme to hydrolyse most oxidized proteins, and the high immunogenicity exhibited suggests a possible involvement of the enzyme in pathogenesis of the disease. PMID- 9228560 TI - Production of anti-peptide antibodies against trypanopain-Tb from Trypanosoma brucei brucei: effects of antibodies on enzyme activity against Z-Phe-Arg-AMC. AB - Anti-peptide antibodies were produced against the cysteine proteinase trypanopain Tb from Trypanosoma brucei brucei and the effects of these antibodies on enzyme activity against carboxybenzoyl (Z)-Phe-Arg-aminomethylcoumarin (AMC) investigated. A peptide was synthesised corresponding to a region of the trypanopain-Tb active site around the active site histidine so that the resulting anti-peptide antibodies specifically targeted the active site of the enzyme. Such antibodies were considered more likely to modulate enzyme activity compared with antibodies directed against other regions of the enzyme. Trypanopain-Tb activity was modulated by rabbit and chicken antibodies produced against both the free and conjugated peptide. Rabbit anti-peptide antibodies enhanced trypanopain-Tb activity by up to 64% at 500 micrograms/ml relative to non-immune antibodies. Chicken antibodies on the other hand, both enhanced (by up to 176% at 500 mg/ml) and inhibited (by up to 85% at 250 mg/ml) trypanopain-Tb activity against Z-Phe Arg-AMC. The nature of the antibody effect depended on the stage during the immunisation protocol at which the antibodies were produced. Chicken antibodies also modulated trypanopain-Tb activity in lysates of T.b. brucei, while rabbit antibodies were only effective against the purified enzyme. Anti-trypanopain-Tb peptide antibodies were thus shown to have the potential to affect trypanopain-Tb activity. PMID- 9228561 TI - Anti-cathepsin D chicken IgY antibodies: characterisation, cross-species reactivity and application in immunogold labelling of human splenic neutrophils and fibroblasts. AB - Hyperexpression, alteration of trafficking and secretion of cathepsin D has been linked with tumour invasion and inflammation. To study these phenomena in a variety of cells large quantities of anti-cathepsin D antibodies and the appropriate immunogen are required. As the human immunogen for studies on human tissue is less easily accessed, antibodies to both human and porcine cathepsin D were raised in chickens, as high levels of antibody may be recovered from egg yolks, and the potential cross-reactivity of the anti-porcine cathepsin D IgY antibody was assessed. This preparation cross-reacted strongly with human cathepsin D, comparing favourably with the reactivity of the chicken antibody to the human immunogen. The necessity for isolating human immunogen can thus be circumvented. The cross-species-reacting chicken IgY was successfully used to localise cathepsin D in immunogold labelling of human tissues. To our knowledge, IgY antibodies have not previously been used by other researchers for this purpose. Application of the cross-reacting antibody to human splenic neutrophils (PMNs) has confirmed the presence of cathepsin D in some granules. Double labelling has shown these to be novel subpopulations of azurophil granules. Cathepsin D may, therefore, be relevant in the invasive and inflammatory activities of PMNs and a target for therapeutic strategies. PMID- 9228562 TI - The Y chromosome in forensic analysis and paternity testing. AB - The male specificity of the human Y chromosome makes it potentially useful in forensic studies and paternity testing, and markers are now available which will allow its usefulness to be assessed in practice. However, while it can be used confidently for exclusions, the unusual properties of the Y mean that inclusions will be very difficult to make: haplotypes are confined within lineages, so population sub-structuring is a major problem, and many male relatives of a suspect will share his Y chromosome. Y haplotyping is most likely to find application in special instances, such as deficiency cases in paternity testing and in the analysis of mixtures of male and female DNA, or in combination with autosomal markers. PMID- 9228563 TI - Evaluation of Y-chromosomal STRs: a multicenter study. AB - A multicenter study has been carried out to characterize 13 polymorphic short tandem repeat (STR) systems located on the male specific part of the human Y chromosome (DYS19, DYS288, DYS385, DYS388, DYS389I/II, DYS390, DYS391, DYS392, DYS393, YCAI, YCAII, YCAIII, DXYS156Y). Amplification parameters and electrophoresis protocols including multiplex approaches were compiled. The typing of non-recombining Y loci with uniparental inheritance requires special attention to population substructuring due to prevalent male lineages. To assess the extent of these subheterogeneities up to 3825 unrelated males were typed in up to 48 population samples for the respective loci. A consistent repeat based nomenclature for most of the loci has been introduced. Moreover we have estimated the average mutation rate for DYS19 in 626 confirmed fatherson pairs as 3.2 x 10( 3) (95% confidence interval limits of 0.00041-0.00677), a value which can also be expected for other Y-STR loci with similar repeat structure. Recommendations are given for the forensic application of a basic set of 7 STRs (DYS19, DYS3891, DYS389II, DYS390, DYS391, DYS392, DYS393) for standard Y-haplotyping in forensic and paternity casework. We recommend further the inclusion of the highly polymorphic bilocal Y-STRs DYS385, YCAII, YCAIII for a nearly complete individualisation of almost any given unrelated male individual. Together, these results suggest that Y-STR loci are useful markers to identify males and male lineages in forensic practice. PMID- 9228564 TI - Chromosome Y microsatellites: population genetic and evolutionary aspects. AB - By means of a multicenter study, a large number of males have been characterized for Y-chromosome specific short tandem repeats (STRs) or microsatellites. A complete summary of the allele frequency distributions for these Y-STRs is presented in the Appendix. This manuscript describes in more detail some of the population genetic and evolutionary aspects for a restricted set of seven chromosome Y STRs in a selected number of population samples. For all the chromosome Y STRs markedly different region-specific allele frequency distributions were observed, also when closely related populations were compared. Haplotype analyses using AMOVA showed that when four different European male groups (Germans, Dutch, Swiss, Italians) were compared, less than 10% of the total genetic variability was due to differences between these populations. Nevertheless, these pairwise comparisons revealed significant differences between most population pairs. Assuming a step-wise mutation model and a mutation frequency of 0.21%, it was estimated that chromosome Y STR-based evolutionary lines of descent can be reliably inferred over a time-span of only 1950 generations (or about 49,000 years). This reduces the reliability of the inference of population affinities to a historical, rather than evolutionary time scale. This is best illustrated by the construction of a human evolutionary tree based on chromosome Y STRs in which most of the branches connect in a markedly different way compared with trees based on classical protein polymorphisms and/or mtDNA sequence variation. Thus, the chromosome Y STRs seem to be very useful in comparing closely related populations which cannot probably be separated by e.g. autosomal STRs. However, in order to be used in an evolutionary context they need to be combined with more stable Y-polymorphisms e.g. base-substitutions. PMID- 9228565 TI - Microvariation at the human D1S80 locus. AB - The minisatellite locus D1S80, (location: 1p35-p36) GenBank sequence accession # D28507), is a variable number of tandem repeat (VNTR) locus with a 16 base pair repeat size. The sequence of the predominant core repeat region and variants of the D1S80 locus were determined to ascertain whether sequence variation or size variation is the cause of altered migration of some D1S80 alleles. A total of 23 alleles from 14 individuals, previously typed based on the number of repeats (i.e. nominal alleles) for the D1S80 locus, were selected for sequence analysis. The individuals were from African American, Caucasian, and Hispanic databases. From these, 18 different repeat unit sequences were observed and arbitrarily designated A-R. Structural relationships between the alleles became more apparent when the arrays of repeat units were divided into common motifs or super-repeat domains. Six motifs ranging from 3 to 9 repeat units were identified. Several of the alleles included repeat arrays which were too diverse to predict an evolutionary relationship, however, there are two general repeat motif arrays and each has some relationship with either the 18 or the 24 repeat allele. The D1S80 allelic polymorphism is primarily due to variation in the number of repeat units and to sequence variation among repeats, however, it can not be ruled out that some rare alleles may be due to insertions or deletions. PMID- 9228567 TI - Influence of the cosmetic treatment of hair on drug testing. AB - An important issue of concern for drug analysis in hair is the change in the drug concentration induced by the cosmetic treatment of hair. The products used for this treatment are strong bases and they are expected to cause hair damage. As a result drugs may be lost from the hair matrix or, under conditions of environmental contamination, be more easily incorporated into the hair matrix. We investigated the effects of cosmetic treatment in vivo by analysing hair samples selected from people who had treated their hair by bleaching or dyeing before sample collection. All of the subjects admitted a similar drug consumption during the time period for which the strands were analysed. Samples were viewed under a microscope to establish the degree of hair damage. Treated and untreated portions from each lock of hair were then selected, separated and analysed by standard detection procedures for cocaine, opiates, cannabinoids and nicotine. In all cases the drug content in hair that had undergone cosmetic treatment decreased in comparison to untreated hair. The majority of the mean differences were in the range of 40%-60% (cocaine, benzoylecgonine, codeine, 6-acetylmorphine and THC COOH). For morphine the mean difference was higher than 60%, and two cases (THC and nicotine) differed by approx. 30%. These differences depended not only on the type of cosmetic treatment, as bleaching produced higher decreases than dyeing, but also on the degree of hair damage i.e. the more damaged the hair, the larger the differences in the concentration levels of drugs. PMID- 9228566 TI - The expression of P-selectin in inflammatory and non-inflammatory lung tissue. AB - An initial attachment of leucocytes to blood vessel walls is mediated by selectins. A feature of adhesion mediated by P-selectin is the "rolling" of leucocytes on the endothelium. The time dependent expression of p-selectin in lung tissue was investigated in five groups of cases with different causes of death: carbon-monoxide and cyanide intoxication (n = 11), drowning (n = 5), hanging (n = 9), pneumonia (n = 13) and polytrauma with blunt thorax trauma (n = 14). In paraffin-embedded archival specimens immunostaining was achieved using an adapted APAAP-immunoperoxidase technique together with a wet autoclave method. P selectin detection was scored by a semiquantitative method evaluating the intensity and incidence of positively stained endothelial cells. The distribution pattern of endothelial P-selectin of blood vessels in cases of pneumonia and septic shock were heterogenius and weak. In one case with lung contusion (survival time 3 h) moderate infiltrates of granulocytes were found near to septal and subpleural hemorrhages. In these inflammatory areas the positive endothelial immunostaining of small vessels was often weaker than in other lung segments or compared to the intensely stained platelets in corresponding vessels. PMID- 9228568 TI - Delayed death after attempted suicide by hanging. AB - In cases of hanging death usually occurs immediately after strangulation and is caused by ischemic cerebral damage due to neck compression in some cases in combination with respiratory obstruction. We report on a case of delayed death 4 days after an attempted suicide by hanging where the individual was conscious and showed no neurological abnormalities. The cause of death was a cerebral infarction following a traumatic thrombosis of the subtotally ruptured carotid arteries. PMID- 9228569 TI - Incorrect identification of a military pilot with international implications. AB - The case is reported of a military pilot shot down in 1986 during a mission whose body was recovered in an advanced state of decomposition and delivered to Italian police in 1989. The first autopsy led to an incorrect identification of the corpse. Because of the advanced decomposed state of the corpse, a correct identification was made only through evaluation of the dental status and radiological examination. The correct evaluation of the specific shape of the amalgam restorations, particularly of those of the first and second inferior right molars, showed agreement with those of the pilot of the plane and not of the co-pilot as was initially stated. PMID- 9228570 TI - Genetic variation at five STR loci in subpopulations living in Turkey. AB - Five short tandem repeat (STR) systems HumVWA, HumTH01, HumCD4, HumF13B and HumFES were investigated in 2 subpopulations living in Turkey (Laz Turks and Kurds). The population genetic data were compared to a Turkish population sample from the Adana area. A closer genetic relationship was found to the Laz Turks than to the Kurdish sample which was also confirmed by phylogenetic tree reconstruction with seven populations from three major ethnic groups (Caucasian, Asian and African). In contrast to the Laz and Adana populations the Kurdish sample showed relatively low heterozygosity values and deviations from Hardy Weinberg equilibrium in four of the five systems. PMID- 9228571 TI - Analysis of the STR myelin basic protein locus in Koreans. AB - Allele und genotype frequencies and the mutation rate of a short tandem repeat locus, the myelin basic protein (MBP) gene, were studied in 973 unrelated Koreans. The alleles were distributed in two discrete regions, one in a high molecular weight region (A) above 190 bp and the other in a low molecular weight region (B) below 150 bp. In a heterozygote, two alleles were found in each region. In region A 13 alleles were found and in region B 7 alleles. Most alleles showed a difference of 4 bp, but three interalleles were found in region A. Allele frequencies in Koreans differed from those reported for Germans and Portuguese. Sets of alleles, one from each region, were linked and transmitted to the offspring. A total of 36 haplotypes and 148 genotypes was identified. In 763 gametes of 550 families, whose parent-child relationship was confirmed using other serological and DNA systems, all alleles were transmitted in a Mendelian fashion, and no mutations were observed. The polymorphism information content (PIC) in Koreans was calculated as 0.833 for region A and 0.718 for region B. The power of discrimination (PD) was 0.959 for region A and 0.901 for region B. No significant deviation from Hardy-Weinberg equilibrium could be observed for this system. PMID- 9228572 TI - An automated procedure for cluster analysis of multivariate satellite data. AB - This paper studies the applicability of available cluster analysis methods on multivariate satellite data. Four different methods are studied, these are the Principal Component Analysis (PCA), K-means, Self Organizing Maps (SOM) and the Adaptive Resonance Theory (ART). Special focus is placed on the usefulness of these cluster analysis methods, that is, if the results generated by these methods are relevant and are of help to the physical analysis of the data. A combined SOM adaptive dynamic K-means procedure suitable for automated cluster analysis is presented. This new method is capable of achieving useful partition of multivariate satellite data which has not previously been studied. PMID- 9228573 TI - Classification of handwritten digits using a RAM neural net architecture. AB - Results are reported on the task of recognizing handwritten digits without any advanced pre-processing. The result are obtained using a RAM-based neural network, making use of small receptive fields. Furthermore, a technique that introduces negative weights into the RAM net is reported. The results obtained on the task of recognizing handwritten digits is comparable with the best performances reported in the literature. PMID- 9228574 TI - Dynamic cell structures for the evaluation of keypoints in facial images. AB - In this contribution Dynamic Cell Structures (DCS network) are applied to classify local image structures at particular facial landmarks. The facial landmarks such as the corners of the eyes or intersections of the iris with the eyelid are computed in advance by a combined model and data driven sequential search strategy. To reduce the detection error after the processing of the sequential search strategy, the computed image positions are verified applying a DCS network. The DCS network is trained by supervised learning with feature vectors which encode spatially arranged edge and structural information at the keypoint position considered. The model driven localization as well as the data driven verification are based on steerable filters, which build a representation comparable with one provided by a receptive field in the human visual system. We apply a DCS based classifier because of its ability to grasp the topological structure of complex input spaces and because it has proved successful in a number of other classification tasks. In our experiments the average error resulting from false positive classifications is less than 1%. PMID- 9228575 TI - Gaussian neural networks for glass bottles inspection: a learning procedure. AB - In glass bottle inspection, the defects detection is of first importance. For online system detection, high speed and robust detection of faults are highly required. Neural networks have recently, and successfully, been applied to fault detection in many manufacturing processes. In this study, a Gaussian neural network, an extension of the RBF network, trained through a competitive algorithm, has been chosen for fault detection. Four parameters extracted from images of the bottles are used as inputs of the network. The number of Gaussian units is adjusted by an informational criterion. Experimental results show that the performance of this network are better than classical parametric and non parametric classifiers. PMID- 9228577 TI - Application of a multi-structure neural network (MSNN) to sorting pistachio nuts. AB - A multi-structure neural network (MSNN) classifier consisting of four discriminators followed by a maximum selector was designed and applied to classification of four grades of pistachio nuts. Each discriminator was a multi layer feed-forward neural network with two hidden layers and a single-neuron output layer. Fourier descriptors of the nuts' boundaries and their area were used as the recognition features. The individual discriminators were trained using a biased technique and a back-propagation algorithm. The MSNN classifier gave an average classification performance of 95.0%. This was an increase of 14.8% over the performance of a multi-layer neural network (MLNN) with similar complexity for classifying the same set of patterns. PMID- 9228578 TI - A hybrid classifier for remote sensing applications. AB - This paper presents a hybrid-unsupervised and supervised-classifier for land use classification of remote sensing images. The entire satellite image is quantized by an unsupervised Neural Gas process and the resulting codebook is labeled by a supervised majority voting process using the ground truth. The performance of the classifier is similar to that of Maximum Likelihood and is only a little worse than Multilayer Perceptions while training and classifying requires no expert knowledge after collecting the ground truth. The hybrid classifier is much better suited to classifications with complex non-normally distributed classes than Maximum Likelihood. The main advantage of the Neural Gas classifier, however, is that it requires much less user interaction than other classifiers, especially Maximum Likelihood. PMID- 9228576 TI - Radar image segmentation using self-adapting recurrent networks. AB - This paper presents a novel approach to the segmentation and integration of (radar) images using a second-order recurrent artificial neural network architecture consisting of two sub-networks: a function network that classifies radar measurements into four different categories of objects in sea environments (water, oil spills, land and boats), and a context network that dynamically computes the function network's input weights. It is shown that in experiments (using simulated radar images) this mechanism outperforms conventional artificial neural networks since it allows the network to learn to solve the task through a dynamic adaptation of its classification function based on its internal state closely reflecting the current context. PMID- 9228579 TI - Neural network visual recognition for automation of the microelectromechanical systems assembly. AB - A neural network visual recognition system is developed. The system is intended for automation of microsystem assembly process. The recognition of microparts is insensitive on their position. This feature is enabled by calculation of the moment properties of the image during preprocessing. The system takes grey-level images and produces recognition code as the output. A feed-forward neural network is used for recognition. Learning of the neural network is performed by combination of standard backpropagation and resilient propagation rule. The system performance is satisfactory with respect to recognition accuracy and recognition time. PMID- 9228580 TI - Growing 3D-SOMs with 2D-input layer as a classification tool in a motion detection system. AB - This paper describes the employment of an 'Adaptive Growing Three-Dimensional Self-Organizing Map' for the classification of images. First a short description of growing SOMs is given and the fundamental advantages are mentioned. Then an extension of the original SOM from two to three dimensions with growing feature is presented. By means of some selected examples the general behavior of the algorithm is illustrated. PMID- 9228581 TI - Using neural networks to automatically detect brain tumours in MR images. AB - Computer vision has been applied to many medical imaging problems with the aim of providing clinical tools to aid medical professionals. We present work being carried out to develop one such system to automatically detect a specific type of brain tumour from head MR images. The tumour under consideration is an acoustic neuroma, which is a benign tumour occurring in the acoustic canals. The hybrid system developed integrates neural networks with more conventional techniques used for computer vision tasks. A database of MR images from 50 patients has been assembled and the acoustic neuromas present in the images have been labelled by hand. Using this data, neural networks (MLPs) have been developed to classify the images at the pixel level to achieve a targeted segmentation. The data used to train and test the MLPs developed, consists of the grey levels of a square of pixels, the pixel to be classified being the centre pixel, together with its global position in the image. The initial pixel level segmentation is refined by a series of conventional techniques. It is combined with an edge-region based segmentation and a morphological operation is applied to the result. This processing produces clusters of adjacent regions, which are considered to be candidate tumour regions. For each possible combination of these regions, features are measured and presented to neural networks which have been trained to identify structures corresponding to acoustic neuromas. Using this approach, all the acoustic neuromas are identified together with three false positive errors. PMID- 9228582 TI - Self-supervised learning in cooperative stereo vision correspondence. AB - This paper presents a neural network model of stereoscopic vision, in which a process of fusion seeks the correspondence between points of stereo inputs. Stereo fusion is obtained after a self-supervised learning phase, so called because the learning rule is a supervised-learning rule in which the supervisory information is autonomously extracted from the visual inputs by the model. This supervisory information arises from a global property of the potential matches between the points. The proposed neural network, which is of the cooperative type, and the learning procedure, are tested with random-dot stereograms (RDS) and feature points extracted from real-world images. Those feature points are extracted by a technique based on the use of sigma-pi units. The matching performance and the generalization ability of the model are quantified. The relationship between what have been learned by the network and the constraints used in previous cooperative models of stereo vision, is discussed. PMID- 9228583 TI - A neural network based artificial vision system for licence plate recognition. AB - This paper presents a neural network based artificial vision system able to analyze the image of a car given by a camera, locate the registration plate and recognize the registration number of the car. The paper describes in detail various practical problems encountered in implementing this particular application and the solutions used to solve them. The main features of the system presented are: controlled stability-plasticity behavior, controlled reliability threshold, both off-line and on-line learning, self assessment of the output reliability and high reliability based on high level multiple feedback. The system has been designed using a modular approach. Sub-modules can be upgraded and/or substituted independently, thus making the system potentially suitable in a large variety of vision applications. The OCR engine was designed as an interchangeable plug-in module. This allows the user to choose an OCR engine which is suited to the particular application and to upgrade it easily in the future. At present, there are several versions of this OCR engine. One of them is based on a fully connected feedforward artificial neural network with sigmoidal activation functions. This network can be trained with various training algorithms such as error backpropagation. An alternative OCR engine is based on the constraint based decomposition (CBD) training architecture. The system has showed the following performances (on average) on real-world data: successful plate location and segmentation about 99%, successful character recognition about 98% and successful recognition of complete registration plates about 80%. PMID- 9228584 TI - Video sequence compression via supervised training on cellular neural networks. AB - In this paper, a novel approach for video sequence compression using Cellular Neural Networks (CNN's) is presented. CNN's are nets characterized by local interconnections between neurons (usually called cells), and can be modeled as dynamical systems. From among many different types, a CNN model operating in discrete-time (DT-CNN) has been chosen, its parameters being defined so that they are shared among all the cells in the network. The compression process proposed in this work is based on the possibility of replicating a given video sequence as a trajectory generated by the DT-CNN. In order for the CNN to follow a prescribed trajectory, a supervised training algorithm is implemented. Compression is achieved due to the fact that all the information contained in the sequence can be stored into a small number of parameters and initial conditions once training is stopped. Different improvements upon the basic formulation are analyzed and issues such as feasibility and complexity of the compression problem are also addressed. Finally, some examples with real video sequences illustrate the applicability of the method. PMID- 9228585 TI - Automatic segmentation and classification of outdoor images using neural networks. AB - The paper describes how neural networks may be used to segment and label objects in images. A self-organising feature map is used for the segmentation phase, and we quantify the quality of the segmentations produced as well as the contribution made by colour and texture features. A multi-layer perception is trained to label the regions produced by the segmentation process. It is shown that 91.1% of the image area is correctly classified into one of eleven categories which include cars, houses, fences, roads, vegetation and sky. PMID- 9228586 TI - Hepatitis delta and beyond. PMID- 9228587 TI - Central venous catheters in patients with AIDS. AB - Central venous catheters (CVCs) for patients with AIDS are at risk of a number of complications including bacterial infections. A 6-year retrospective review was undertaken of the records of the 33 patients (42% infected by injection drug use (IDU)) who received intravenous therapy both in hospital and at home via CVCs. Twenty-eight per cent of 53 insertions suffered a complication, the commonest of which was a pneumothorax (8%). The post insertion complication rate was 0.98/100 catheter days (cd). Thrombotic occlusion (0.15/100 cd) was the commonest non septic event while sepsis was overall the commonest event (0.69/ 100 cd) of which half were considered serious (0.33/100 cd). The most frequently isolated organisms were Staphylococci spp. (71%). The median time to an exit site infection was 59 days and to serious catheter sepsis 86 days. Infection did not differ significantly with age, gender, transmission risk activity or catheter type although Portacaths had the lowest rate of infection (0.33/100 cd). The median survival of the 53 CVCs was 88 days although if the temporary catheters were excluded it was 118 days. Kaplan-Meier estimates of survival analysis revealed 55%, 32% and 19% of all the CVCs surviving 3, 6 and 12 months respectively. Our experience suggests that home intravenous therapy and previous IDU does not preclude the use of CVCs although further research is needed on reducing the infection-related complications of such therapy. PMID- 9228588 TI - Decrease in CD4 lymphocyte counts with rest; implications for the monitoring of HIV infection. AB - Measurement of the CD4 lymphocyte count is widely used as a prognostic marker and guide for the institution of antiretroviral therapy in patients infected with HIV (human immunodeficiency virus). CD4 counts are known to fluctuate with strenuous physical activity and diurnal variation but there is no information on the effects of rest or normal daily activity. We investigated the effects of rest on the absolute CD4 lymphocyte count in 20 healthy laboratory workers. Blood samples were obtained in 20 subjects upon arrival in the laboratory (CD4 0), following 30 and 60 min rest (CD4 30 and CD4 60 respectively) and 8 h into a normal working day (CD4 8). A significant decrease in the CD4 lymphocyte count was observed following 60 min rest; mean CD4 count at 0 min 1060 x 10(6)/L, mean CD4 count at 60 min 660 x 10(6)/L (P = 0.0017). These results demonstrate a significant effect of rest on CD4 lymphocyte counts in healthy volunteers. This biological variation may be important in HIV-infected patients and needs to be addressed by further studies. PMID- 9228589 TI - An actual use comparison of condoms meeting Australian and Swiss standards: results of a double-blind crossover trial. AB - The performance of condoms in actual use has been poorly researched in the past, especially in comparing condoms that met different quality control standards as indicated by laboratory testing. The present study used a double-blind crossover design to compare the performance of 2 types of condoms in actual use; one that met the Australian and International Organization for Standardization (ISO) standards for condom quality and one that met the more stringent Swiss Quality Seal requirements. Ninety-two men recruited from Metropolitan Melbourne completed a self-report diary sheet after each condom was used which assessed the performance of the condom and the conditions under which it was used. From a total of 1917 condom uses, there was an overall breakage risk of 2.7%. The breakage risk ratio (Australian/ISO:Swiss) for all types of use was 1.16 (95% confidence interval 0.68-1.99). When subanalyses by method of entry were performed, the condoms meeting the Swiss standard appeared to fare better than the Australian/ ISO standards for anal sex (RR = 4.84, 95% CI 1.07-21.8, P = 0.022), while the opposite was the case for vaginal sex (RR = 0.74, 95% CI 0.35 1.53, P = 0.41). The result for anal use was statistically significant at the 5% level, despite being based on fewer condom trials than that for vaginal use, but this result needs to be replicated. Although the participants appeared representative of the general male population in Melbourne in the age bracket 18 46 years, there was a significant history of condom usage reported. This may have influenced the risk of breakage. PMID- 9228591 TI - Determinants of HIV seroconversion in injection drug users during a period of rising prevalence in Vancouver. AB - To identify determinants of HIV seroconversion among injection drug users (IDUs) during a period of rising prevalence, a case-control investigation was conducted. Cases were IDUs with a new positive test after 1 January 1994, and a negative test within the prior 18 months. Controls required 2 negative tests during the same period. Subjects completed a questionnaire on demographic, psychosocial, and behavioural factors. Eighty-nine cases and 192 controls were similar with respect to gender, age, ethnicity and inter-test interval. Multivariate analyses of events during the inter-test interval showed borrowing syringes (adj. OR = 2.96; P < 0.006), unstable housing (adj. OR = 2.01; P = 0.03) and injecting > or = 4 times daily (adj. OR = 1.71; P = 0.06) to be independently associated with seroconversion. Protective associations were demonstrated for sex with opposite gender (adj. OR = 0.36; P = 0.001) and tetrahydrocannabinol use (adj. OR = 0.41; P = 0.001). There is a need to evaluate programmes dealing with addiction, housing and the social underpinnings of risk behaviours in this population. PMID- 9228590 TI - Are acetowhite lesions of the cervix correlated to the presence of Epstein-Barr virus DNA? AB - The purpose of this study was to investigate whether Epstein-Barr virus (EBV) is associated with acetowhite lesions of the portio cervix, demonstrating koilocytosis and/or cervical intraepithelial neoplasia (CIN) I-III. The study group comprised 37 women admitted to the Department of Gynaecology and Obstetrics, Sahlgrenska University Hospital, Goteborg because of pathological colposcopy or cytology of the portio cervix. Colposcopically, all exhibited acetowhite lesions of the portio cervix. Cells were sampled with a cytobrush for examination for EBV and human papillomavirus (HPV) DNA by polymerase chain reaction (PCR) and a biopsy was taken for histopathology. Biopsies from 5 patients positive for EBV by PCR in cervical cell samples were examined by an in situ hybridization technique for EBER (Epstein-Barr virus encoded RNA), RNAs expressed in latent EBV infection. The control group consisted of women attending the Department of Dermato-Venereology at the same hospital for STD check-up. These had a normal cytology and no signs of acetowhiteness of the portio cervix. In the study group, EBV DNA was found in 30% and HPV DNA in 51%. In the control group 57% exhibited EBV DNA and 23% HPV DNA. EBV was not found to be a predictive factor in the development of koilocytosis and/or CIN I-III. HPV was a predictive factor in acetowhite, koilocytotic lesions. No expression of EBER was found in the 5 biopsies examined by in situ hybridization. PMID- 9228592 TI - Sexually transmitted diseases in Estonia: past and present. AB - The present survey covers historical events in Estonia during the era of the USSR regime and the era after independence as regards incidence of sexually transmitted diseases (STDs). The diagnostical methods used as the reporting system are presented. Reasons for the increased incidence of traditional venereal diseases such as gonorrhoea and syphilis are discussed. The importance of migration of prostitutes from Russia is also considered. PMID- 9228593 TI - Levels of dangerousness: an analysis of Irish dentists' clinical responses to HIV seropositive patients. AB - Work conducted in the mid-1980s suggested that dentists in the Republic of Ireland felt uncomfortable about treating patients who were HIV-seropositive. It seemed that with greater understanding of the behaviour of the virus, dentists' attitudes would be modified accordingly. The aim of this study was to assess the reactions of Irish dentists to the treatment of HIV-positive patients. All dentists currently on the register were sent a questionnaire to assess their knowledge, attitudes and clinical behaviours. Over 60% of dentists responded. The study highlights dentists' responses to HIV infection suggesting that they are fearful of the virus. They perceive HIV as dangerous to themselves, their other patients and their practice. The findings indicate that attitudes relating to dangerousness remain a significant obstacle in the treatment of this patient group. PMID- 9228595 TI - Cerebral abscesses in a patient with AIDS caused by methicillin-resistant Staphylococcus aureus (MRSA) PMID- 9228594 TI - Antimicrobial self medication in patients attending a sexually transmitted diseases clinic. AB - One of the health education messages given in sexually transmitted disease (STD) control is patients' adopting appropriate health seeking behaviour. This includes reporting to health facilities for appropriate diagnosis and treatment. In parts of the world where STD aetiologic agents have assumed resistance to commonly used antimicrobials, this is important. The antimicrobial self medication practices of 764 patients attending an STD clinic in a developing country were studied. Seventy-four and a half per cent admitted to self medication before reporting to the clinic. The antibiotics taken in inappropriate dosages were purchased over the counter, given by friends or were 'left-overs' from previous medications. In the fight to control STD spread as a means of reducing the incidence of HIV/AIDS, indiscriminate use of antimicrobials needs to be guarded against. PMID- 9228596 TI - Metastatic Crohn's disease causing a vulval mass and involving the cervix. PMID- 9228597 TI - Heterosexually transmitted HIV infection in the UK. PMID- 9228599 TI - Azithromycin in gonorrhoea. PMID- 9228598 TI - Do women with pelvic inflammatory disease receive adequate treatment? PMID- 9228600 TI - Arsenical pessaries in the treatment of metronidazole-resistant Trichomonas vaginalis. PMID- 9228601 TI - Flat-panel x-ray detector using amorphous silicon technology. Reduced radiation dose for the detection of foreign bodies. AB - RATIONALE AND OBJECTIVES: The authors evaluate a new flat-panel x-ray detector (FD) with respect to foreign body detection and reduction of radiation dose compared with screen-film radiography. METHODS: Flat-panel x-ray detector is based on amorphous silicon technology and uses a 1 k x 1 k photo-detector matrix with a pixel size of 143 x 143 microns and 12-bit digital output. A thallium dotted cesium iodide scintillation layer converts x-rays into light. An ex vivo experimental model was used to determine the detectability of foreign bodies. Foreign bodies with varying sizes were examined: glass with and without addition of lead, bone, aluminium, iron, copper, gravel fragments, and graphite. Four hundred observation fields were examined using conventional radiography (speed, 400; system dose: 2.5 microGy) as well as FD with a simulated speed of 400, 800, 1200, and 1600, corresponding to a detector dose of 2.5 microGy, 1.25 microGy, 0.87 microGy, and 0.625 microGy, respectively. Four independent radiologists performed receiver operating characteristic analysis of 8000 observations. RESULTS: Flat-panel x-ray detector with a simulated speed of 400 was significantly superior (P = 0.012) to screen-film radiography (speed, 400). At a simulated speed of 800 and 1200 FD yielded results equivalent to screen-film radiography. Flat-panel x-ray detector was significantly inferior to screen-film radiography at a simulated speed of 1600 (P = 0.012). CONCLUSIONS: Flat-panel x ray detector technology allows significant reduction in radiation dose compared with screen-film radiography without loss of diagnostic accuracy. PMID- 9228602 TI - Optimal technique factors for magnification mammography. AB - RATIONALE AND OBJECTIVES: Use of small focal spots with low x-ray tube currents may result in very long exposure times and thus result in motion blur in magnification mammography. The authors investigated the reduction in exposure time with increasing x-ray tube kVp and the corresponding decrease in perceived visibility of low-contrast objects in phantom images. METHODS: Exposure times required to radiograph an RMI 156 phantom in a magnification geometry were measured as a function of x-ray tube kVp when operated under automatic exposure control. Magnification images of the RMI 156 phantom were obtained at x-ray tube voltages ranging from 28 to 34 kVp. Five radiology residents ranked the visibility of two borderline fibers and six borderline microcalcification specks using a 5-point scale ranging from excellent to barely visible. RESULTS: Between 28 and 34 kVp, the density of the RMI phantom images was nearly constant with a mean value of 1.32 +/- 0.04. Increasing the x-ray tube voltage from 28 kVp to 34 kVp reduced the exposure time from 1.27 seconds to 0.66 seconds. Image quality at 30 and 32 kVp was not significantly worse than that achieved at 28 kVp. Increasing the x-ray tube voltage to 34 kVp, however, resulted in a statistically significant (P < 0.001) deterioration in the relative visibility of fibers and microcalcification specks. CONCLUSIONS: Magnification mammography performed at 32 kVp will decrease exposure times significantly and result in a microcalcification and fiber visibility that is similar to that achieved at 28 kVp. PMID- 9228603 TI - Phosphorus-31 magnetic resonance spectroscopy studies of pig spinal cord injury. Myelin changes, intracellular pH, and bioenergetics. AB - RATIONALE AND OBJECTIVES: Phosphorus-31 (31P) nuclear magnetic resonance (NMR) spectroscopy was used to monitor changes in phosphocreatine (PCr), adenosine triphosphate (ATP), inorganic phosphate (Pi), intracellular pH (pHi), and free magnesium in the in vivo pig spinal cord after injury. METHODS: Phosphorus-31 NMR spectra were acquired from healthy (n = 4) and injured pig spinal cords (n = 8) under in vivo conditions using a 4.7-tesla spectrometer. Spinal cords were injured by dropping a 20-g weight from 20 cm onto the surgically exposed cord surface. RESULTS: In vivo spectra of injured cords revealed a reduction in ATP, PCr, pHi, and an increase in Pi. In addition, a broad resonance that is likely to arise from myelin phospholipids was reduced significantly after injury. CONCLUSIONS: Phosphorus-31 NMR can be used to follow in vivo changes in high energy phosphates after injury and may have the potential to follow changes in myelin structure. This technique may prove important in the study of myelin breakdown after secondary, nonreversible spinal cord injury. Changes in high energy phosphates and pHi did not seem to parallel these putative changes in myelin structure. PMID- 9228604 TI - Color-coded duplex sonography of experimentally induced multilevel stenosis. Evaluation of poststenotic Doppler spectrum. AB - RATIONALE AND OBJECTIVES: The authors evaluated the influence of a proximal arterial stenosis on the poststenotic doppler spectrum of a second, distal stenosis and determined duplex parameters, which permitted description of the severity of the distal stenosis. METHODS: Moderate (ie, 50%) and severe (ie, 90%) stenoses of the distal aorta and the distal iliac arteries of 10 pigs were created surgically and characterized by angiography. All possible combinations of moderate and severe stenoses were examined. The Doppler spectrum was depicted in the distal iliac and distal femoral arteries of both limbs and analyzed by use of maximum and minimum flow velocity (Vmax, Vmin), acceleration index (modified Handa index), acceleration time, and pulsatility index. RESULTS: In cases of moderate as well as severe proximal stenosis, acceleration index, acceleration time, and pulsatility index of the poststenotic curve of the distal stenosis were significantly reduced (P < 0.05). Independent of the severity of the proximal stenosis, differentiation of moderate as well as severe distal stenosis was possible (P < 0.05) with these parameters. CONCLUSIONS: Despite interference of spectral curves in proximal and distal stenosis, duplex sonography enabled the differentiation of experimental aortic iliac multilevel stenosis. PMID- 9228605 TI - Evaluation of effect of treatment for invasive bladder cancer by ultrasonography with intra-arterial infusion of carbon dioxide microbubbles. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to evaluate the diagnostic usefulness of the ultrasonography with intra-arterial infusion of carbon dioxide microbubbles (CO2) for invasive bladder cancer. METHODS: Twelve patients with muscle-invading bladder cancer who were treated by concurrent radiotherapy and intra-arterial infusion of daily low dose of cisplatin using an implanted infusion port were included. A total of 30 studies was performed during the treatment to evaluate the visualization of the tumor and effect of the treatment compared with conventional ultrasonography, computed tomography, or cystoscopy. RESULTS: Satisfactory visualization of the tumor in CO2 ultrasonography was obtained in all patients, in particular in those with flat tumor or prostatic invasion. The enhancement effect of CO2 on the tumor, which was maintained well in the late period of the treatment, made possible evaluation of the therapeutic effect. With respect to the evaluation of local response, disagreement between clinical response and the evaluation of CO2 ultrasonography was observed in two patients with definite differentiation between wall edema and residual tumor after treatment being difficult. CONCLUSIONS: Carbon dioxide ultrasonography is easy to perform in patients treated with arterial infusion therapy using an implanted infusion port and provides practical information in evaluating therapeutic effect. PMID- 9228606 TI - Magnetic resonance imaging in the diagnosis of acute injured distal tibiofibular syndesmosis. AB - RATIONALE AND OBJECTIVES: This study assessed the diagnostic potential of magnetic resonance imaging for the evaluation of the tibiofibular syndesmosis. METHODS: A total of 38 patients with an acute ankle trauma and clinical suspicion of a syndesmotic tear were prospectively studied with conventional plain film radiography and magnetic resonance imaging. Magnetic resonance imaging studies included plain T1-weighted (T1-w) and T2-weighted (T2-w) sequences and contrast enhanced T1-w sequences 0 to 3 days after trauma. All images were read by two independent radiologists before surgical intervention. Sensitivity and specificity were determined for the two observers and the concordance of the two observers were calculated using the interobserver analysis (Kappa-Test). Intraoperative inspection (n = 21) revealed rupture of the anterior tibiofibular ligament (ATIF) in 15 patients, intact ATIF in 6 patients, and intact posterior tibiofibular ligament (PTIF) in 21 cases. Clinical and follow-up examinations revealed an intact syndesmotic complex in another 17 patients. RESULTS: Primary diagnostic criteria for diagnosing a ligamentous tear included tibiofibular diastasis in conventional plain films; nonvisualization of the ATIF; an abnormal course, a wavy, irregular contour of the ligament; increased signal intensity of the ligament in T2-w sequences, in plain T1-w sequence, and marked enhancement in T1-w after contrast. Important secondary signs were defined as joint fluid in the tibiofibular space and prolapse of interspace fat. Highest diagnostic accuracy was achieved if three or more diagnostic criteria could be visualized. Both readers performed best with the enhanced T1-weighted and the T2-weighted images in transverse orientation. The interobserver analysis resulted in high concordance: Kappa = 0.9 (confidence interval: 0.76 to 1.00) for all patients, and in Kappa = 0.76 (confidence interval: 0.45 to 1.0) for surgically treated patients. CONCLUSIONS: Magnetic resonance imaging of the syndesmotic complex is a highly sensitive and specific tool for the pretherapeutic-evaluation of syndesmotic injury. PMID- 9228607 TI - Gray-scale liver enhancement in VX2 tumor-bearing rabbits using BR14, a new ultrasonographic contrast agent. AB - RATIONALE AND OBJECTIVES: We evaluated the potential of BR14, a novel gas-based echo contrast agent, for gray-scale liver imaging and VX2 tumor detection/delineation in the conventional and harmonic imaging modes. METHODS: A single dose (0.2 mL/kg) of BR14 was administered to eight VX2 tumor-bearing rabbits. Imaging was performed with an ATL-HDI 3000. The B-mode gray levels were analyzed by videodensitometry in areas selected in the liver tissue, tumors, aorta, and the portal vein from frames recorded up to 1 hour after injection. The tumors were further assessed using subjective scores just before and after contrast injection, as well as 15 minutes later in both conventional gray-scale and harmonic imaging modes. RESULTS: Gray-scale enhancement of the liver tissue was detectable soon after BR14 injection and remained at a high level even after clearance of the agent from the blood stream. On precontrast, of 42 observed tumors, 4 were perfectly detected (4/42). After injection of BR14, the number rose to 10/20 in the vascular phase and 12/20 in the delayed phase. In harmonic imaging, the corresponding numbers were even higher, 17/22 in the vascular phase and 18/22 in the delayed phase. The number of tumors perfectly delineated increased after BR14 from 1/42 to 3/20 in the vascular phase and 5/20 in the delayed phase. In the harmonic mode, 9 of 22 tumors were perfectly delineated in the vascular phase and 10 of 22 in the delayed phase. CONCLUSIONS: BR14 is a promising new agent for the study of tissue perfusion and in particular for liver imaging. It shows not only strong, but also persistent gray scale enhancement of the parenchyma but not of the tumors. The increased conspicuousness allows considerable improvements in tumor detection and tumor delineation in conventional imaging and even more so in harmonic imaging. PMID- 9228608 TI - Contrast-enhanced magnetic resonance urography. First experimental results with a polymeric gadolinium bloodpool agent. AB - RATIONALE AND OBJECTIVES: The authors investigated the feasibility of contrast enhanced excretory magnetic resonance urography to visualize the nonobstructed urinary tract with a macromolecular gadolinium-based bloodpool agent. METHODS: Excretory magnetic resonance imaging was performed in seven pigs using a T1 weighted three dimensional fast-field-echo sequence before and up to 120 minutes after administration of a gadolinium bloodpool prototype agent. RESULTS: During the first 15 minutes after injection, the urographic effect was predominantly poor. Visualization of the entire urinary tract was excellent in four pigs and incomplete but satisfactory in three 105 minutes after injection. Furosemide application was tested in one case, which improved image quality effectively. Corresponding to the physiological excretion rate, signal measurements in the renal parenchyma revealed a gradual decrease of the initially distinct contrast enhancement. CONCLUSIONS: T1-weighted contrast-enhanced magnetic resonance urography using a polymeric gadolinium bloodpool allows detailed visualization of the normal urinary tract, while information about the excretory function is obtained simultaneously. However, application of a diuretic seems to be essential to prevent lengthy examination duration. PMID- 9228609 TI - Effect of contrast medium on corpus cavernosum smooth muscle. AB - RATIONALE AND OBJECTIVES: Changes in contractility of corpus cavernosum (CC) smooth muscle caused by radio contrast medium may result in misinterpretations of cavernosography used diagnostically in erectile dysfunction. METHODS: The authors investigated the direct effect of various contrast media on rabbit CC smooth muscle tissue strips in an in vitro model by adding contrast medium to the tissue in a perfusion bath and recording the resulting contractions. Glucose addition was used as control. RESULTS: Application of high-osmolar, ionic contrast medium diatrizoate-induced CC smooth muscle contractions of 57% of the control potassium chloride (124 mM) induced contractions. The low-osmolar (862 mOsm/kg) nonionic monomer contrast medium, iohexol, and the iso-osmolar (300 mOsm/kg) nonionic dimer contrast medium, iodixanol, elicited contractions of 34% and 36% of the potassium chloride control contractions, respectively. High- and Iso-osmolar glucose solutions caused contractions of 51%, 38%, and 24% of the control, respectively. Cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) regulate CC smooth muscle contractions. These are influenced by different drugs including phosphodiesterases (PDEs), forskolin, and 3 morpholinsydnonimine hydrochloride (SIN-1). The nonspecific PDE inhibitors papaverine (0.1 mM) and theophylline (1 mM) reduced the contrast medium-induced contractions to 66% and 69%, respectively. The specific PDE inhibitor milrinone (0.1 mM) reduced the contractions to 69%; 0.1 mM forskolin and SIN-1 reduced the contractions to 34% and 41%, respectively. CONCLUSIONS: Contrast medium induces CC smooth muscle contractions, depending mainly on the osmolality of the solution. The contractions are reduced but not abolished by elevating the intracellular cAMP and cGMP concentrations. The clinical applications in cavernosography are discussed. PMID- 9228610 TI - Genomics via optical mapping. II: Ordered restriction maps. AB - In this paper, we describe our algorithmic approach to constructing ordered restriction maps based on the data created from the images of population of individual DNA molecules (clones) digested by restriction enzymes. The goal is to devise map-making algorithms capable of producing high-resolution, high-accuracy maps rapidly and in a scalable manner. The resulting software is a key component of our optical mapping automation tools and has been used routinely to map cosmid, lambda and BAC clones. The experimental results appear highly promising. PMID- 9228611 TI - Hardness of flip-cut problems from optical mapping. AB - Optical mapping is a new technology for constructing restriction maps. Associated computational problems include aligning multiple partial restriction maps into a single "consensus" restriction map, and determining the correct orientation of each molecule, which was formalized as the Exclusive Binary Flip Cut (EBFC) Problem in (Muthukrishnan and Parida, 1997). Here we prove that the EBFC problem, as well as a number of its variants, are NP-complete. Therefore, they do not have efficient, that is, polynomial time solutions unless P = NP. PMID- 9228612 TI - Finding genes in DNA with a Hidden Markov Model. AB - This study describes a new Hidden Markov Model (HMM) system for segmenting uncharacterized genomic DNA sequences into exons, introns, and intergenic regions. Separate HMM modules were designed and trained for specific regions of DNA: exons, introns, intergenic regions, and splice sites. The models were then tied together to form a biologically feasible topology. The integrated HMM was trained further on a set of eukaryotic DNA sequences and tested by using it to segment a separate set of sequences. The resulting HMM system which is called VEIL (Viterbi Exon-Intron Locator), obtains an overall accuracy on test data of 92% of total bases correctly labelled, with a correlation coefficient of 0.73. Using the more stringent test of exact exon prediction, VEIL correctly located both ends of 53% of the coding exons, and 49% of the exons it predicts are exactly correct. These results compare favorably to the best previous results for gene structure prediction and demonstrate the benefits of using HMMs for this problem. PMID- 9228613 TI - A six-point condition for ordinal matrices. AB - Ordinal assertions in an evolutionary context are of the form "species s is more similar to species x than the species y" and can be deduced from a distance matrix M of interspecies dissimilarities (M[s, x] < M[s, y]). Given species x and y, the ordinal binary character cxy of M is defined by cxy(s) = 1 if and only if M[s,x] < M[s, y], for all species s. In this paper we present several results concerning the inference of evolutionary trees or phylogenies from ordinal assertions. In particular, we present. A six-point condition that characterizes those distance matrices whose ordinal binary characters are pairwise compatible. This characterization is analogous to the four-point condition for additive matrices. An optimal O(n2) algorithm, where n is the number of species, for recovering a phylogeny that realizes the ordinal binary characters of a distance matrix that satisfies the six-point condition. An NP-completeness result on determining if there is a phylogeny that realizes k or more of the ordinary binary characters of a given distance matrix. PMID- 9228614 TI - Recognizing circular decomposable metrics. AB - Circular decomposable metrics (CDMs) have been used in phylogenetic studies. The fastest algorithm for recognizing a CDM runs in time O(n5), given an n x n table of pairwise distances. We give an O(n2) time algorithm for this problem. PMID- 9228615 TI - Path costs in evolutionary tree reconstruction. AB - This paper describes a dynamic programming algorithm to solve a family of problems in the reconstruction of evolutionary trees from protein sequence data, that of constructing "minimal" colorings. This dynamic programming formulation can be modified to efficiently enumerate the number of minimal colorings and thereby be used to calculate the average cost of any given edge, where the average is taken over the entire set of minimal colorings. An extension of our dynamic programming formulation allows for the calculation of average path costs amongst all minimal colorings. our results resolve questions raised in (Hendy and penny, 1987); in particular, we develop polynomial time procedures to find the minimum, maximum, and average (expected) cost of an edge, and more generally of a path, for a minimal coloring. Our algorithm is distinguished in its flexibility to address further distribution and statistical questions relating to the minimal colorings. Furthermore, the more general concept of calculating statistics describing the set of optimal solutions may be of interest in other combinatorial problems. PMID- 9228616 TI - On a Mirkin-Muchnik-Smith conjecture for comparing molecular phylogenies. AB - A conjecture of Mirkin, Muchnik, and Smith is answered affirmatively which connects the inconsistency function, a biologically meaningful similarity/dissimilarity measure for a gene tree and a species tree, to the mutation cost function, a combinatorial measure based on the mapping of trees. A linear-time algorithm for computing the mutation cost function is also derived from the conjecture. PMID- 9228617 TI - An efficient statistic to detect over- and under-represented words in DNA sequences. AB - In this note, we point out a very efficient statistic to detect over- and under represented words in DNA sequences, when Markov chain models are used to represent the sequences. This statistic is missing from the recent review done on this important problem and appears to be a better measure of rarity and abundance of words in DNA sequences. PMID- 9228618 TI - Statistical modeling, phylogenetic analysis and structure prediction of a protein splicing domain common to inteins and hedgehog proteins. AB - Inteins, introns spliced at the protein level, and the hedgehog family of proteins involved in eucaryotic development both undergo autocatalytic proteolysis. Here, a specific and sensitive hidden Markov model (HMM) of protein splicing domain shared by inteins and the hedgehog proteins has been trained and employed for further analysis. The HMM characterizes the common features of this domain including the position where a site-specific DNA endonuclease domain is inserted in the majority of the inteins. The HMM was used to identify several new putative inteins, such as that in the Methanococcus jannaschii klbA protein, and to generate a multiple sequence alignment of sequences possessing this domain. Phylogenetic analysis suggests that hedgehog proteins evolved from inteins. Secondary and tertiary structure predictions suggest that the domain has a structure similar to a beta-sandwich. Similarities between the serine protease cleavage mechanism and the protein splicing reaction mechanism are discussed. Examination of the locations of inteins indicates that they are not inserted randomly in an extein, but are often inserted at functionally important positions in the host proteins. A specific and sensitive HMM for a domain present in klbA proteins identified several additional bacterial and archaeal family members, and analysis of the site of insertion of the intein suggests residues that may be functionally important. This domain may play a role in formation of surface associated protein complexes. PMID- 9228619 TI - Withdrawal of antihypertensive medications. AB - BACKGROUND: Pharmacologic treatment of hypertension reduces risks of stroke, congestive heart failure, renal failure, and mortality, but whether medications, once begun, need to be continued for life is uncertain. METHODS: Several search strategies on MEDLINE using key words "medication," "withdrawal," "discontinuance," and "therapy" in several combinations, nested within "hypertension," were not productive. Accordingly, articles known to the authors and citations within them were reviewed. A survey of a random sample of members of the New York Academy of Family Practice was conducted to ascertain current practice of practicing physicians. RESULTS: Eighteen studies of antihypertensive medication withdrawal were located and all were reviewed. In 12 trials average success rates of 40.3 percent after 1 year of follow-up and 27.7 percent after 2 years were achieved. In six studies limited to elderly patients, an average success rate of 26.2 percent was obtained for periods of 2 or more years. The trials, however, were heterogeneous in design, patient selection criteria, and follow-up. The survey of family physicians indicated that 79.1 percent attempt withdrawal of antihypertensive medications in hypertensive patients whose blood pressure is controlled and who are without symptoms from medication. CONCLUSIONS: We conclude that successful withdrawal of antihypertensive medications can have substantial benefits with few or no adverse consequences and might be successful in about one third of patients. Additional research is required to substantiate rates of successful medication withdrawal, to define the best method of withdrawing medications, and to delineate characteristics of patients in whom withdrawal is most likely to succeed. PMID- 9228620 TI - On-site colposcopy services in a family practice residency clinic: impact on physician test-ordering behavior, patient compliance, and practice revenue generation. AB - BACKGROUND: Using colposcopy as a model, we examined the impact of introducing a new diagnostic technology into the ambulatory primary care setting. METHODS: Records of patients with abnormal findings on Papanicolaou smears were reviewed from three study periods: 1 year before, 1 year after, and 5 years after initiation of on-site colposcopy services. Data analyzed include physician management decisions, site of colposcopic service, and patient compliance. Practice revenue estimates were based upon patterns of physician management and patient compliance found during each study period. RESULTS: Management of low grade squamous intraepithelial lesions varied during each study period. By period 3, however, most patients were undergoing colposcopy (P = 0.03). High-grade squamous intraepithelial lesions were uniformly managed with colposcopy during all study periods (P < 0.001). Introduction of on-site colposcopic services resulted in a rapid shift to the on-site location for evaluation of low-grade squamous intraepithelial lesions and a more gradual shift to the on-site location for evaluation of high-grade squamous intraepithelial lesions. Patient compliance was not affected by the introduction of on-site services. On-site colposcopy resulted in a nearly 100 percent transfer of revenue to the practice, but the economic benefit was quite modest. CONCLUSIONS: Although offering on-site colposcopy services might have had some impact on physician management of low grade squamous intraepithelial lesions, the lack of benefit regarding patient compliance, the relatively small patient volume for this procedure, and its modest impact on practice revenue cause us to question the value of including colposcopy in everyday practice. PMID- 9228621 TI - Dietary supplement users: demographics, product use, and medical system interaction. AB - BACKGROUND: Dietary supplements-defined as vitamins and minerals, herbal products, tissue extracts, proteins and amino acids, and other products-are purchased to improve health and prevent disease. Little has been published, however, about the characteristics of either the products or the people who use them. METHODS: Consecutive customers visiting two health food stores during a 15 day period were interviewed by telephone. They were asked about their use of dietary supplements, demographics, and their use of the established health care system. RESULTS: Of the 194 customers contacted, 136 (70.1 percent) completed the survey. Respondents took a total of 805 supplements, most often to prevent a health problem (84.3 percent). Herbal products were most commonly used. Garlic, ginseng, and Ginkgo biloba were the herbs most frequently used. Fifty products were found to have previously reported toxicities, including vitamin A, which 9 customers were taking in megadoses. Most customers were white (94.1 percent), female (75.7 percent), had at least 1 year of college education (70.6 percent), had health insurance (95.6 percent), and had a regular physician (85.3 percent). CONCLUSION: Most of the dietary supplements consumed appear to be safe, but 50 of 805 had previously reported toxicities including megadoses of vitamin A. Garlic, ginseng, and Ginkgo biloba were the most commonly ingested herbs, and the medical literature supports their effectiveness for some conditions in humans. Customers of two health food stores had average to above-average education and took dietary supplements to stay healthy. They used the conventional health care system but did not typically consult their physician about dietary supplements. The pattern of use suggests that physicians might not be adequately addressing preventive and wellness issues in discussions with their patients. Furthermore, physicians might need to learn about dietary supplements so they can communicate with patients about them. PMID- 9228622 TI - The National Health Service Corps: rural physician service and retention. AB - BACKGROUND: The National Health Service Corps (NHSC) scholarship program is the most ambitious program in the United States designed to supply physicians to medically underserved areas. In addition to providing medical service to underserved populations, the NHSC promotes long-term retention of physicians in the areas to which they were initially assigned. This study uses existing secondary data to explore some of the issues involved in retention in rural areas. METHODS: The December 1991 American Medical Association (AMA) Masterfile was used to determine the practice location and specialty of the 2903 NHSC scholarship recipients who graduated from US medical schools from 1975 through 1983 and were initially assigned to nonmetropolitan counties. We used the AMA Masterfile to determine what percentage of the original cohort was still practicing in their initial county of assignment and the relation of original practice specialty and assignment period to long-term retention. RESULTS: Twenty percent of the physicians assigned to rural areas were still located in the county of their initial assignment, and an additional 20 percent were in some other rural location in 1991. Retention was highest for family physicians and lowest for scholarship recipients who had not completed residency training when they were first assigned. Retention rates were also higher for those with longer periods of obligated service. Substantial medical care service was provided to rural underserved communities through obligated and postobligation service. Nearly 20 percent of all students graduating from medical schools between 1975 and 1983 who are currently practicing in rural counties with small urbanized populations were initially NHSC assignees. CONCLUSIONS: Although most NHSC physicians did not remain in their initial rural practice locations, a substantial minority are still rural practitioners; those remaining account for a considerable proportion of all physicians in the most rural US counties. This study suggests that rural retention can be enhanced by selecting more assignees who were committed to and then completed family medicine residencies before assignment. PMID- 9228623 TI - Anxiety disorders in elderly patients. AB - BACKGROUND: Late-life anxiety disorders, commonly seen in primary care settings, can coexist with other medical and psychiatric illnesses. A variety of effective treatment options is available for these patients. METHODS: MEDLINE was searched for articles published from 1970 to 1996 using the key words "anxiety," "elderly," "aged," "geriatric," "panic," "obsessive-compulsive," "phobia," and "generalized anxiety disorder." Studies of patients older than 65 years were reviewed. RESULTS: Generalized anxiety disorder, phobias, panic disorder, and obsessive-compulsive disorder are the most common late-life anxiety problems seen by primary care physicians. Patients with these disorders complain of diffuse multisystem symptoms, motor restlessness, and such physiologic symptoms as tachycardia or tachypnea. Comorbid illnesses include depression, alcoholism, drug use, and multisystem disease. Behavioral strategies to address anxiety include an open discussion of the issue, an anxiety diary, psychosocial support, and cognitive-behavioral techniques. Pharmacologic strategies include carefully monitored benzodiazepine, buspirone, or antidepressant therapy. CONCLUSIONS: Clinical trials of all anxiety interventions are needed for elderly primary care patients to clarify further whether findings from mixed-age population studies are generalizable to the elderly. PMID- 9228624 TI - Cervical pregnancy--a forgotten entity in family practice. AB - BACKGROUND: Cervical pregnancy is a rare form of ectopic pregnancy that is associated with considerable maternal morbidity and a high mortality rate if early diagnosis and treatment are not carried out in a timely fashion. METHODS: The current medical literature was reviewed by searching MEDLINE files from 1985 to 1996, using the key words "ectopic pregnancy" and "cervical pregnancy." Older articles were accessed from cross-reference of the more recent publications. RESULTS: The incidence of cervical pregnancy is 1 in 2400 deliveries and represents less than 1.0 percent of all ectopic pregnancies. No clear cause of cervical pregnancy has been described, and criteria for the clinical, pathologic, and sonographic diagnosis have been well established. The most common clinical complaint is painless vaginal bleeding. Routine transvaginal sonography early on allows for conservative management and avoids adverse outcomes. Methotrexate administered systemically and by intra-amniotic instillation are the therapeutic options of choice. Gestational age and the presence or absence of fetal cardiac activity are major prognostic factors for its success. CONCLUSION: Earlier diagnosis of cervical pregnancy using sonography and conservative management of this condition have reduced considerably the morbidity and mortality associated with this rare form of ectopic pregnancy and have helped preserve a woman's future fertility. PMID- 9228625 TI - Pneumothorax following acupuncture. PMID- 9228626 TI - Morning rounds and the search for evidence-based answers to clinical questions. PMID- 9228627 TI - Beyond signing the death certificate. PMID- 9228628 TI - Warning--the physician's clinical judgement can be hazardous to your health: withdrawing drugs in patients with high blood pressure. PMID- 9228630 TI - Perimortal care: comments on three vignettes about death. PMID- 9228629 TI - The role of procedures in family practice: is there a right answer? PMID- 9228631 TI - Care of the premature infant. PMID- 9228632 TI - Anesthesia for neonatal circumcision. PMID- 9228633 TI - Morphological changes induced by short pulse hydrogen fluoride laser radiation on dental hard tissue and restorative materials. AB - BACKGROUND AND OBJECTIVE: The potential benefits of the effects of lasers on dental tissues have yet to be realized but may be brought closer through the availability of a suitable laser. The objective of this project is to examine the surface morphological changes resulting from hydrogen fluoride (HF) laser radiation on tooth and restorative material surfaces. STUDY DESIGN/MATERIALS AND METHODS: A hydrogen fluoride laser emitting at 2.9 microns is used to interact with a range of dental hard tissue and restorative materials. The surface morphological changes induced by 100 mJ pulses of < 1 microsecond duration is studied using a SEM. RESULTS: The irradiated surfaces displayed microstructures similar to those of a mechanically fractured surface with no evidence of melting. CONCLUSION: This study suggests that tissue is removed by microexplosion, leaving a surface free from thermal damage with surface characteristics that would appear to facilitate the adhesion of restorative materials. PMID- 9228634 TI - Control of hypertrophic scar growth using selective photothermolysis. AB - BACKGROUND AND OBJECTIVE: Previous studies have shown a clinical improvement of hypertrophic scars (HS) after treatment with a pulsed dye laser. The objective of this study was to investigate the effects of variations in pulse wavelength and energy density on HS tissue using human HS implanted in athymic mice. STUDY DESIGN/MATERIALS AND METHODS: Small pieces (approximately 1 mm3) of HS tissue were implanted into athymic mice and allowed to grow for 5 days. The implant site was then exposed to a single 450 microseconds pulse, and implant growth and histology were monitored for an additional 12 days. Laser wavelength and energy density ranges tested were 585-600 nm and 2-10 J/cm2, respectively. RESULTS: Using a wavelength of 585 nm, laser treatment inhibited implant growth by 70% at 6 J/cm2 and 92% at 10 J/cm2, respectively. The inhibitory effect decreased as the laser wavelength was increased from 585 to 600 nm. A widespread destruction of the implant microvasculature with a minor effect on surrounding extracellular matrix at the highest light dose were observed. CONCLUSION: Pulsed laser treatment inhibits HS implant growth in nude mice. This effect is likely mediated by selective photo-thermolysis of the implant microvasculature. PMID- 9228635 TI - Use of Er:YAG laser for benign skin disorders. AB - BACKGROUND AND OBJECTIVE: The Er:YAG laser is of special interest in dermatology and cosmetic surgery since it ablates and cuts tissue with surgical precision with minimal collateral thermal damage due to the wavelength of the Er:YAG radiation (2,940 nm), which is strongly absorbed by liquid water in tissue. The study was designed to establish optimal laser parameters for treating various skin disorders. STUDY DESIGN/MATERIALS AND METHODS: Sixty-four patients were treated for benign skin disorders: seborrhoic warts, plane warts, milia, xanthelasma palpebrarum, hidradenoma, chloasma, senile lentigo, epidermal naevi, actinic keratosis, fibroepithelial papillomata, scars. The lesions were irradiated with single pulse laser energies 100-1,000 mJ, repetition rates 2-10 Hz, and spot diameters 2-8 mm. RESULTS: Epidermis was effectively removed on a layer-by-layer basis. For the ablation, energy densities higher than 2.5 J/cm2 were required. If bleeding appeared, the hemostatic effect was achieved by irradiating the bleeding surface with few Er:YAG laser pulses of lower power density (0.5-1.5 J/cm2). Healing was excellent and without apparent scarring. CONCLUSION: It was established that Er:YAG laser with properly selected parameters offers a tool for tissue ablation and/or coagulation. The Er:YAG laser was found to be a perfect option for effective treatment of benign skin disorders. PMID- 9228636 TI - Development of a smart holmium:YAG laser lithotriptor. AB - BACKGROUND AND OBJECTIVE: The purpose of this study was to develop a feedback control system for the pulsed holmium:YAG medical laser that enhances tissue selectivity and safety by discriminating between soft and hard biological tissue such as urinary and biliary calculi and bone. STUDY DESIGN/MATERIALS AND METHODS: The ability to discriminate is achieved by monitoring prompt laser-induced visible/NIR photoemissions via retrograde transmission over the laser delivery fiber in conjunction with a developed detection algorithm. RESULTS: Experimental data are presented for a system that employs this discrimination scheme with an electro-optic shutter for rapid intrapulse feedback control of holmium laser based lithotripsy procedures. The results demonstrate the feasibility of a lithotriptor that can deliver 1 J per pulse to calculi yet limit errant discharges to surrounding urinary tract tissue to < or = 0.1 J. CONCLUSION: Based on animal tissue safety data, the laser margin of safety is improved by an order of magnitude. PMID- 9228637 TI - Holmium:YAG laser and pulsed dye laser: a cost comparison. AB - BACKGROUND AND OBJECTIVE: This study was designed to evaluate the relative cost effectiveness of the Holmium:YAG laser and the pulsed dye laser for the treatment of ureteral calculi. Cost containment is a priority for every health care facility. As a result, the staff of the Lutheran Medical Center (Wheat Ridge, CO) looked at alternative ways to provide quality laser treatment of ureteral stones. As part of our study, the laser committee offered the Holmium:YAG laser to urologists for ureteral lithotripsy. Previously, the pulsed dye laser was rented for ureteral calculi on a per case basis at $1,500. A hospital processing fee was added to this cost, resulting in a total charge of $1,638 to the patient. Our organization owns a Holmium:YAG laser and uses it primarily in orthopedics. STUDY DESIGN/MATERIALS AND METHODS: Two ureteral lithotripsy cases were performed and compared. One case used the Holmium:YAG for ureteral lithotripsy; the other procedure used the pulsed dye laser. A cost analysis was performed after the procedures. RESULTS: The data indicated a significant difference in cost between the two lasers. Approximately $1,000 was eliminated when using the Holmium:YAG laser. CONCLUSION: A cost savings of $15,000 per year would be realized if 15 cases were performed. The Holmium:YAG laser also can be used on cystine calculi, a procedure for which the pulsed dye laser is ineffective. The potential for ureteral injury exists. When using the Holmium:YAG laser, appropriate training is required. Due to this risk, not all urologists will use the Holmium:YAG laser. We also found a positive correlation between the proficiency of the urologists' laser skills and overall cost effectiveness. PMID- 9228639 TI - Importance of using rigorous statistical methods to analyze low energy laser experimental data: Part two. AB - BACKGROUND AND OBJECTIVE: Numerous authors have reported successful alteration of peripheral nerve action potential characteristics through application of low energy laser irradiation (LELI). The statistical analysis that accompanies many of these reports frequently does not account for the special nature of the data generated in typical LELI experiments. The objective of this study was to evaluate the application of repeated measures linear regression techniques to the analysis of this type of data. Issues of analyzing raw versus normalized data, proper accounting for correlation between measurements, and discrete time point hypothesis testing were addressed. STUDY DESIGN/MATERIALS AND METHODS: The data analyzed in this work were generated from an experiment in which in vitro frog sciatic nerves were irradiated with a helium-neon laser using a variety of treatment protocols. Compound action potential (CAP) amplitude, latency, depolarization rate, and repolarization rate were recorded at 1-minute intervals for 135 minutes for each nerve. Laser-induced changes in CAP parameters were analyzed using various repeated measures linear regression models. RESULTS: The findings of statistical significance were highly dependent on the rigor of the regression model applied. Application of the same regression model to raw and normalized data produced different findings of significance. Determination of significant contrasts was highly dependent on how well the regression model accounted for the correlation between repeated measurements made on the same nerve. In general, models that failed to account adequately for this correlation produced more findings of significant contrasts than increasingly rigorous models. Finally, discrete time point hypothesis testing on normalized data can suggest improper statistical conclusions if the proper correlation structure is not applied to the data set. CONCLUSION: Linear regression analysis offers advantages over discrete time point hypothesis testing in the analysis of highly correlated serial data of this type. Trends in the behavior of the measured parameters are evident, rigorous accounting for correlation between measurements is facilitated, and hypothesis testing is highly flexible. PMID- 9228638 TI - In vitro frog sciatic nerve as a peripheral nerve model for studies of the mechanism of action of low energy lasers: Part one. AB - BACKGROUND AND OBJECTIVE: There have been numerous reports of modulation of peripheral nerve action potential characteristics through application of low energy laser irradiation (LELI), although no mechanism has yet been advanced to explain these observations. In order to investigate the mechanism of LELI effects in peripheral nerve tissue, a well-characterized, reliable, and robust peripheral nerve preparation is required. The objective of this study was to evaluate the in vitro frog sciatic nerve as a candidate model for future LELI mechanism studies. MATERIALS AND METHODS: Following 60-minute baseline recordings of compound action potential (CAP) amplitude, latency, depolarization rate, and repolarization rate, helium-neon (HeNe) laser irradiation (632 nm, 15 min, 1-7 J, 44-320 J/cm2) was delivered to one of two sites on the nerve. Laser-induced changes in CAP parameters were analyzed during irradiation and for 60 minutes post-irradiation using a repeated measures linear regression model. RESULTS: In the treatment group that received 7 J of HeNe energy over the recording electrode, CAP latency increased relative to nonirradiated controls during the postirradiation period. No other treatment group demonstrated laser-induced changes in CAP characteristics at any time during the experiment. CONCLUSION: HeNe irradiation demonstrated limited ability to alter the CAP under these conditions. As such, the in vitro frog sciatic nerve is an inappropriate model for mechanism of action studies. PMID- 9228640 TI - Comparison of 810 nm and 1064 nm wavelengths for interstitial laser photocoagulation in rabbit brain. AB - BACKGROUND AND OBJECTIVE: This laboratory animal study is a comparison of Nd:YAG 1064 nm and diode 810 nm laser wavelengths in brain interstitial laser photocoagulation (ILP). Specific goals were to identify potential complications and physical characteristics of the thermal damage at both wavelengths prior to undertaking a clinical trial in humans. STUDY DESIGN/MATERIALS AND METHODS: A total of 41 ILP illuminations were performed in vivo in the brains of 33 anesthetized rabbits using plane-cut fiber tips implanted directly or through catheters, and diffusing fiber tips. Delivered powers ranged from 1.1 to 4.2 W. Exposures ranged from 300 to 900 s. Survival ranged from 0 to 48 h. Experiments were performed in animals with and without VX-2 brain tumors. RESULTS: Thermal damage from 1.1 W at 810 nm was similar to that from 1.6 W at 1064 nm, but more pronounced. With plane-cut fiber tips, there was a greater propensity for severe physical effects (smoke, charring, bubbling, surface damage) at 810 nm than at 1064 nm, yet hemorrhage, thrombosis and vapor dissemination were observed at both wavelengths, in both normal brain and tumor. CONCLUSIONS: For ILP in brain, 1064 nm may be better suited than 810 nm, although both are questionable with plane cut-fiber tips. Compactness and portability may be the only valid reasons for using laser diodes operating around 810 nm. At 1064 nm, the power delivered from plane-cut fiber tips should be less than 1.5 W, necessitating long exposures, or else an open catheter should be used. Fiber tips with distributed emission may be preferred, provided structural integrity can be maintained. PMID- 9228641 TI - Effects of heterogeneous absorption of laser radiation in biotissue ablation: characterization of ablation of fat with a pulsed CO2 laser. AB - BACKGROUND AND OBJECTIVE: Physicians encounter several clinical situations in which fat must be removed. In this study, the characterization of fat ablation produced by a pulsed CO2 laser is reported. STUDY DESIGN/MATERIALS AND METHODS: An RF excited 800 microns pulsed CO2 laser operating at 10.6 microns was used to ablate fresh porcine fat. The heat of ablation and ablation threshold were determined using a mass loss technique. Absorption coefficients for fat and dermis were determined by attenuated total reflection spectroscopy. RESULTS: Threshold radiant exposure and heat of ablation for fat were calculated from the mass loss measurements to be 1.05 J/cm2 and 2.4 J/cm3, respectively. The absorption coefficients of fat and dermis at 10.6 microns were 250 and 780 cm-1, respectively. Pulsed CO2 laser ablation of fat caused ejection of fat droplets, which ignited after high fluence pulses. CONCLUSION: A pulsed CO2 laser can effectively ablate fat with a threshold fluence and efficiency comparable to other soft tissues. Our data suggest that fat ablation occurs primarily through the ejection of intact fat particles via the explosive vaporization of intervening water "lakes". PMID- 9228642 TI - Degradation-induced transmission losses in silica optical fibers. AB - BACKGROUND AND OBJECTIVE: The interaction of surgical optical fibers with tissue has been studied. STUDY DESIGN/METHODS AND MATERIALS: Fibers (600 microns) were lased in chicken and beef tissue using a Nd:YAG laser from 5 to 50 W in both cw and pulsed modes. RESULTS: With longer lasing and higher power, larger transmission loss and degradation (burn-in) of the fiber tip, occurred. This degradation converts the Nd:YAG laser power to heat and leads to further energy loss. During contact lasing, tissue and blood adhere to the fiber tip surface limiting laser transmission, desiccating, and eventually destroying adhering tissues. Such tissue residues create high power densities and temperatures at the tip, which then cause a variety of degradation processes to be initiated. CONCLUSION: "Burned-in" fibers do not photocoagulate; rather they incise tissue. With continued lasing, thermal shock, chemical, and mechanical breakdown of the fiber leads to failure of the fiber tip and the spalling of glass fragments into the tissue bed. PMID- 9228643 TI - Perspectives of coronary excimer laser angioplasty: multiplexing, saline flushing, and acoustic ablation control. AB - BACKGROUND AND OBJECTIVE: Bubble formation, pressure wave generation, and cavitations constitute major factors influencing the outcome of clinical Excimer laser angioplasty. Thus, the rationale of this study was to determine the extent of pressure waves occurring during excimer laser ablation and to discuss possibilities that allow a less traumatic plaque removal in the coronary circulation. STUDY DESIGN/MATERIALS AND METHODS: Conventional and experimental Xenon-Chlorid-Excimer lasers emitting light at a wave-length of 308 nm and a pulse duration of 115 ns were used for testing of signals. Whereas the conventional excimer laser light source transmits light through all fibers of a 1.7 mm laser catheter simultaneously, the prototype excimer laser divides the laser beam into several areas of uniform energy fluence by scanning the beam from one section to the other using the intermission between two laser discharges. Hydrophones consisting of piezoelectric films detected the acoustic signals, which were obtained on normal arterial wall and atherosclerotic plaque. RESULTS: Multiplexing decreases maximum pressures for both normal arterial wall and calcified plaque significantly, whereas pressure rise time remains comparable. During ablation of pure blood, a linear increase of peak pressures of 1 MPa at 10 mJ/mm2 to 7.5 MPa at 50 mJ/mm2 is found. Contrast media intensifies the extent of pressure wave formation. At 20 mJ/mm2, 60% contrast media added to blood results in an increase of maximum pressures from 1.5 MPa up to 5 MPa. Dilution with saline solution is effective; however, high concentrations of > 90% are required to achieve a significant pressure wave reduction. CONCLUSION: Peak pressures of several thousand kPa occur during excimer laser ablation of contrast media, blood, calcified plaque, and normal arterial wall in a decreasing order. Multiplexing and saline flushing are capable of reducing the intensity of the generated acoustic signals during tissue ablation. It has to be taken into consideration, however, that high concentrations of saline solution are necessary to achieve a significant reduction of peak pressures. PMID- 9228644 TI - The Er:YAG laser in ear surgery: first clinical results. AB - BACKGROUND AND OBJECTIVE: The goal of this study was to gain experiences about the possibilities and limits of the Er:YAG laser for ear operations. STUDY DESIGN/PATIENTS AND METHODS: Eighty-three ear operations were performed with the aid of an Er:YAG laser: 32 stapedotomies, 15 tympanoplasties type III, 10 tympanoplasties type I, 18 ear operations in cholesteatoma, and 8 removals of hyperostosis in the outer ear canal. RESULTS: The Er:YAG laser facilitated stapedotomies and removal of hyperostosis from the outer ear canal. In cases of beginning cholesteatoma, the Er:YAG laser allowed matrix removal of the ear ossicles left in situ. Furthermore, in tympanoplasty it was possible to achieve an osteosynthesis of the auditory ossicles, which was done for the first time during this study. No hearing loss attributable to laser dose was found during postoperative hearing tests. CONCLUSION: The Er:YAG laser seems to become a useful tool in middle ear surgery. PMID- 9228645 TI - In vitro measurements of cytotoxic effects of 193 nm and 213 nm laser pulses at subablative fluences. AB - BACKGROUND AND OBJECTIVE: The frequency-quintupled q-switched Nd:YAG laser is being studied as an alternative to the ArF excimer laser for photorefractive procedures. The present report describes two experiments comparing biologic effects of these laser devices. STUDY DESIGN/MATERIALS AND METHODS: Bovine corneas were irradiated with subablative laser pulses in liquid nitrogen and analyzed by electron paramagnetic resonance spectroscopy to assess free radical production. Aqueous bacterial suspensions were irradiated with low-intensity laser pulses and assayed for cell survival. RESULTS: Electron paramagnetic resonance spectra were very similar in both amplitude and shape for exposure at the two wavelengths. Bacterial survival was markedly less for 213 nm irradiation than 193 nm exposure and displayed a different dependence on cumulative exposure. CONCLUSION: Free radical production by 213 nm laser exposure is quite comparable to that seen previously for 193 nm irradiation. However, cell lethality appears to be significantly greater at the longer ultraviolet wavelength. This difference may contribute to complications observed after corneal photoablation with the 213 nm device. PMID- 9228646 TI - Estrogenic and progestagenic activities of physiologic and synthetic androgens, as measured by in vitro bioassays. AB - Estrogenic activities of testosterone (T) and 5a-dihydrotestosterone (DHT) were detected and measured by using their specific stimulatory effects on alkaline phosphatase (AP) activity in human endometrial adenocarcinoma cells of the Ishikawa Var-1 line. These two physiologic androgens were able to induce, at microM concentrations, estrogenic effect believed to be mediated by the estrogen receptor (ER) since the antiestrogens ICI-164384 and 4-hydroxytamoxifen (OHTam), but not the antiandrogens hydroxyflutamide (OHFl) or cyproterone acetate (CPA), reversed that effect. By using another in vitro bioassay, based on the progestin specific stimulation of AP activity in cells of the T47D human breast cancer line, progestagenic activity was detected and measured in T, DHT and three synthetic androgens: nandrolone (19-nortestosterone). 7 alpha-methyl 19 nortestosterone (MENT) and mibolerone (7 alpha, 17 alpha-dimethyl 19 nortestosterone) (DMNT). While progestagenic effects of T and DHT required relatively high concentrations (microM levels), the synthetic androgens stimulated AP activity at nM or pM levels. These effects seem to be mediated by the progesterone receptor (PR), since they are completely abolished by the antiprogestins RU-486, ZK-98299 and ZK-112993, but not by the antiandrogen OHFl. These simple in vitro bioassays, expressing biological effects of the test compounds in human cells in culture, revealed dual or multiple hormonal activities coexisting in a single compound and provide quantitative information of considerable pharmacological importance concerning the complex actions of drugs. PMID- 9228647 TI - Role of eicosanoid inhibition of ischemia reperfusion injury: intact and isolated rat heart studies. AB - The effects of FPL-55712 (leukotriene receptor antagonist) and OKY-046 (thromboxane synthase inhibitor) were studied on ischemic and reperfused myocardium using in vivo and in vitro rat heart paradigms. The in vivo studies included production of regional ischemia by coronary artery ligation for 72 h followed by reperfusion. Macroscopic methods using nitro blue tetrazolium (NBT) staining were employed to measure infarct size. For in vitro studies, isolated rat hearts were perfused by Langendorff's technique. Regional ischemia was produced for 60 min followed by reperfusion. Coronary outflow and creatine phosphokinase (CPK) release in the coronary effluent were measured. Both these parameters showed a correlation with the duration of reperfusion. In isolated heart studies, FPL-55712 reduced CPK release and improved coronary outflow significantly, whereas OKY-046 did not show any beneficial effect. In intact heart studies, FPL-55712 did not reduce the infarct size, whereas OKY-046 decreased the myocardial infarct to a significant extent. The present study demonstrates a definite involvement of leukotrienes and thromboxanes in reperfusion injury. Inability of FPL-55712 to reduce infarct size may be due to the very short half-life of the drug. Lack of efficacy of OKY-046 in isolated studies suggests the involvement of platelet-derived thromboxanes in the reperfusion injury as these are not available in isolated heart preparations. PMID- 9228648 TI - Reversal of doxorubicin resistance in multidrug resistant melanoma cells in vitro and in vivo by dipyridamole. AB - The occurrence of multidrug resistance (MDR) decreases the clinical utility of several anticancer agents, including doxorubicin (DOX). A transmembrane efflux pump, P-glycoprotein (P-gp), is frequently implicated in the development of MDR in tumor cells. Dipyridamole (DP), a clinically used antiplatelet drug, enhances the cytotoxicity of the anticancer drugs affected by MDR. Although this aspect has been studied extensively in cell culture models, the effectiveness of DP to overcome multidrug resistance has not been investigated using in vivo models of multidrug-resistant solid tumors. Therefore, the objective of this study was to evaluate the role of DP in the reversal of resistance to DOX in tumor-bearing mice in the context of its anti-MDR activity in vitro. For this purpose, drug sensitive murine melanoma cells (B16V) and their DOX-selected MDR variant, B16VDXR cells, were used. In vitro, the reversal of DOX resistance of B16VDXR cells by DP was determined using clonogenic assays, and the influence of DP on the transport of DOX was evaluated by measurement of steady-state accumulation as well as efflux of DOX in B16VDXR cells. Antitumor activity of different treatments was assessed by monitoring tumor growth. Pharmacokinetics of DOX, with or without DP, were evaluated in C57BL/6 mice bearing B16V or B16VDXR tumors. DP produced a 6.4-fold reversal of resistance to DOX in vitro; this was accompanied by an increase (3.6-fold) in the steady-state intracellular accumulation of DOX and a marked reduction in the efflux of DOX from B16VDXR cells. Furthermore, a linear correlation was observed between the EC50 values and the steady-state intracellular levels of DOX in the multidrug-resistant cells. In the in vivo experiments, similar growth patterns were seen for the DOX alone and the DOX+DP groups for B16V tumors. The results with B16VDXR tumors were in sharp contrast. The DOX+DP treatment caused a significant delay in the growth of B16VDXR tumors compared to treatment with DOX alone or controls. DP did not alter the plasma pharmacokinetics of DOX in C57BL/6 mice but resulted in a significant increase in the intratumoral accumulation of DOX. PMID- 9228649 TI - Stobadine pretreatment enhances glutathione peroxidase activity in the heart of irradiated mice. AB - The effect of pretreatment with stobadine (a novel drug with cardioprotective properties) on the activity of glutathione peroxidase was studied in the heart of mice after Co60 irradiation. Exposure to 6.5 Gy caused significant decrease in the activity of the enzyme (p < 0.01). Treatment with stobadine (70.07 mg/kg) 1 or 2 h before irradiation resulted in activity enhancement in comparison with the nonpretreated irradiated group (p < 0.01). We conclude that the radical scavenging mechanism may be involved in the protection exerted by stobadine. The results are in agreement with those obtained by the micronucleus test. PMID- 9228650 TI - Effects of NIK-247 on cholinesterase and scopolamine-induced amnesia. AB - The effects of NIK-247 on cholinesterase, scopolamine-induced amnesia and spontaneous movement were examined and compared with those of the well-known cholinesterase inhibitors tacrine and E-2020. NIK-247, tacrine and E-2020 all strongly inhibited acetylcholinesterase (AChE) in human red blood cells (IC50s = 1.0 x 10(-6), 2.9 x 10(-7) and 3.7 x 10(-8) M, respectively). In addition, NIK 247 and tacrine, but not E-2020, strongly inhibited butyrylcholinestrase (BuChE) in human serum. All three drugs produced mixed inhibition of AChE activity. Moreover, the inhibitory effect of NIK-247 on AChE was reversible. All compounds at 0.1-1 mg/kg p.o. significantly improved the amnesia induced by scopolamine (0.5 mg/kg s.c.) in rats performing a passive avoidance task. The three compounds at 1 and 3 mg/kg p.o. did not significantly decrease spontaneous movement by rats. These findings suggest that NIK-247 at a low dose (0.1-1 mg/kg p.o.) improves scopolamine-induced amnesia but does not affect spontaneous movement. The findings suggest that NIK-247 may be a useful drug for the treatment of Alzheimer's disease. PMID- 9228651 TI - Effects of intracerebroventricular injection of histamine on memory deficits induced by hippocampal lesions in rats. AB - The influence of bilateral hippocampal lesions on active avoidance response was studied in rats, as well as the effect of intracerebroventricular (i.c.v.) injection of histamine on memory deficits caused by hippocampectomy. Retardation of learning acquisition was produced by lesioning of the bilateral dorsal hippocampus in active avoidance response. Memory retention was also impaired by hippocampectomy. Although locomotor activity and rearing behavior measured by open-field test increased after hippocampal lesions, there was no relation between impairment of learning and increase in exploratory behavior. I.c.v. injection of histamine and i.p. injection of histidine resulted in an improvement of memory deficits (not only learning acquisition but also memory retrieval) induced by hippocampal lesions in rats. Histamine contents of the hippocampus and hypothalamus decreased after hippocampectomy, and a decrease in histamine contents of both areas was restored by histamine (i.c.v.) and histidine (i.p.) injection. In addition, a close relationship was found between decrease in response latency of avoidance response and an increase in histamine content of the hippocampus and hypothalamus after histamine injection. PMID- 9228652 TI - Bropirimine as neoadjuvant therapy decreases residual disease and expression of markers PCNA and TGF-beta 1 in a rat orthotopic prostate adenocarcinoma. AB - The role of bropirimine in prostate cancer remains unexplored. To address the efficacy of this immune modulator as neoadjuvant therapy we utilized the orthotopic placement of the Dunning AT-3 tumor. 2.4-2.6 x 10(6) Dunning AT-3 cells were injected into the ventral prostates of 50 Copenhagen X Fischer rats. Animals were then divided into 5 groups consisting of: 1) untreated controls; 2) those treated with ventral prostatectomy alone (performed 10-12 days following tumor cell inoculation); 3) those treated with ventral prostatectomy plus bropirimine (10 mg/kg) on postimplantation days 1, 3, 5, 10 and 11; 4) those treated with ventral prostatectomy plus bropirimine (100 mg/kg), at the same schedule; and 5) those treated with ventral prostatectomy plus bropirimine (500 mg/kg), at the same schedule. Animals were sacrificed 10 days after prostatectomy, autopsied, and residual disease was weighed. Prostate weights upon removal following neoadjuvant treatment and residual disease remaining after 20 22 days were expressed in grams (g). Following prostatectomy, mean prostate weights were: Group 2, 0.67 +/- 0.11; Group 3, 0.53 +/- 0.11; Group 4, 0.54 +/- 0.12; Group 5, 0.44 +/- 0.09. The effect of bropirimine was significant (p = 0.0001) by multiple regression analysis. In addition, mean residual tumor weights (expressed in grams) after 20-22 days were: Group 1, 12.7 +/- 1.9; Group 2, 6.7 +/- 4.8; Group 3, 5.2 +/- 5.9; Group 4, 3.8 +/- 3.5; and Group 5, 2.8 +/- 3.5. The effect of bropirimine was not significant (p = 0.07) by multiple regression analysis. However, prostatectomy alone, by Student's test, significantly (p = 0.04) reduced residual mean tumor weights by 47% and the additional effect of bropirimine upon residual disease was significant (p = 0.038) if a Chi-square analysis is applied. Finally, a multivariate analysis of the overall effect of bropirimine in rats treated with prostatectomy was significant (p = 0.002). The effect of bropirimine on expression of proliferating cell nuclear antigen (PCNA) and transforming growth factor beta 1 (TGF-beta 1) was also evaluated immunohistochemically and expression of both tumor markers was significantly reduced (p < 0.05). We conclude that bropirimine may have a role as a neoadjuvant therapy when combined with prostatectomy. PMID- 9228653 TI - Effects of oral treatment with sustained release morphine tablets on hypothalamic pituitary-adrenal axis. AB - Morphine is suggested to influence immune function by activation of hypothalamic pituitary-adrenal axis. Thus, we investigated 8 pain patients prior to and during prolonged oral treatment with 30-240 mg of sustained release morphine for plasma concentrations of adrenocorticotropic hormone and serum concentrations of cortisol. Results revealed that pain patients at basal status had elevated cortisol concentrations. Hormone concentrations measured after 1, 4 and 12 weeks of morphine treatment were significantly decreased, not normalized, but at very low values. Since these data, even in the absence of clinical symptoms, might have been indicative for adrenal insufficiency, a corticotropin-releasing hormone test was performed in 2 patients. After injection of 100 micrograms of human corticotropin-releasing hormone. ACTH and cortisol concentrations increased sufficiently. In conclusion, even though low hormonal concentrations were observed in pain patients during morphine treatment, pituitary and adrenal stimulation of the endocrine axis remained intact. PMID- 9228654 TI - Delineation of the border zone of ischemic rabbit myocardium by a technetium labeled nitroimidazole. AB - Delineation of viable ischemic myocardium is an important problem in nuclear cardiology. To determine the feasibility of using a technetium-labeled nitroimidazole as an indicator of ischemic myocardium at risk of infarction, we characterized the distribution of a 2-nitroimidazole-derivatized PnAO ligand and its 99mTc complex, 99mTcO(PnAO)-1-CH2-(2NI) (BMS-181321) in the ischemic territory of the left anterior descending (LAD) coronary artery of the rabbit. In preliminary experiments, the performance of 14C-deoxyglucose (14C-2DG) and 14C misonidazole was assessed relative to apparent regional relative myocardial blood flow (rMBF) indicated by 99mTc-teboroxime using double-label autoradiography in the rabbit LAD occlusion model. After demonstrating that 14C-2DG and 14C misonidazole are selectively retained in the lateral border of the ischemic territory, BMS-181321 was co-injected intravenously, with either 14C-2DG or 14C misonidazole, 20 min after LAD occlusion. In a separate experiment, 99mTcO(PnAO) 6-CH3, a complex with the same lipophilicity (log k' 0.26 vs. 0.31) as BMS-181321 but which lacks the 2NI moiety, was co-injected with 14C-2DG. After 30 min, the rabbits were sacrificed and 14C/99mTc autoradiograms were obtained from the same tissue sections. The autoradiograms revealed that BMS-181321 was retained with the same microregional distribution as both 14C-2DG and 14C-misonidazole in the border zone of the ischemic LAD territory. The selective retention of BMS-181321 depends on the presence of the nitroimidazole group, since 99mTcO(PnAO)-6-CH3 has a uniformly low myocardial distribution in contrast to the enhanced uptake of co injected 14C-2DG. These data demonstrate that BMS-181321 is selectively retained in hypoxic myocardium and demarcates the ischemic border zone in a manner similar to 14C-2DG and 14C-misonidazole. PMID- 9228655 TI - Synthesis, estrogen receptor binding, and tissue distribution of [18F]fluorodoisynolic acids. AB - Doisynolic acids, D-ring seco-steroids derived from alkaline fusion of estrones, are hormonal curiosities: Their binding affinity for the estrogen receptor is low (ca. 1-2% that of estradiol), but their in vivo potency is high and they have a long duration of action. To study the in vivo behavior of the doisynolic acids, we prepared fluorine-substituted analogs of both trans-doisynolic acid (with the natural 14 alpha-hydrogen configuration, trans-FDA) and the more active cis doisynolic acid (with the unnatural 14 beta-hydrogen configuration, cis-FDA) from estrone and 14 beta-estrone, respectively. Modification of the D-ring haloform cleavage approach of Meyers allowed us to introduce fluorine (or fluorine-18) on the carbon atom derived from C-16 in the estrones. Fluorine substitution had little effect on the estrogen receptor binding affinity of the doisynolic acids. Tissue distribution of the fluorodoisynolic acids (trans-[18F]FDA and cis [18F]FDA) was unusual and very different from that of typical, high-affinity ligands for the estrogen receptor. At 1-3 h in immature female rats, trans [18F]FDA shows low and rather nonselective uptake in the principal estrogen target tissue (uterus) and slow clearance. By contrast, cis-[18F]FDA shows high uptake in nearly all tissues, with significant uterine uptake that continues to increase over the 1-6-h period. The uterine uptake of this isomer was blocked at the later times by a sufficiently high dose of unlabeled cis-FDA. After administration of the trans-[18F]FDA, a more polar metabolite slowly accumulates in the blood. The cis-[18F]FDA, however, showed no apparent metabolism, with 84% of the blood activity at 5 h assigned as the unmetabolized radioligand. After 5 h, only limited clearance from blood, liver, and kidneys has occurred. No metabolite from this isomer accumulates in the uterus. Although fluorodoisynolic acids will not be useful breast-tumor imaging agents, their behavior was found to be interesting as it deviates from that of other F-18 estrogens. Further long term studies of cis-doisynolic acid, labeled with tritium, may be needed to explicate fully its unusual distribution properties and high in vivo activity. PMID- 9228656 TI - In vitro and in vivo evaluation of structure-stability relationship of 111In- and 67Ga-labeled antibody via 1B4M or C-NOTA chelates. AB - 2-(p-isothiocyanatobenzyl)-1,4,7-triazacyclononane-1,4,7-triacetic acid (C-NOTA) or 2-(p-isothiocyanatobenzyl)-6-methyl-diethylenetriamine pentaacetic acid (1B4M) was conjugated to monoclonal antibody T101 (IgG2a), radiolabeled with 111In or 67Ga and then purified through size-exclusion HPLC. 111In 1B4M-T101 and 67Ga C NOTA-T101 were stable in in vitro serum at 37 degrees C. In contrast, 111In C NOTA-T101 and 67Ga 1B4M-T101 were unstable. The biodistribution in normal mice reflected the instability of the metal complex; the less-stable 111In C-NOTA conjugate left less tracer in blood, but more in liver and kidney whereas the less-stable 67Ga 1B4M conjugate left less tracer in blood, but more in bone. The biodistribution data suggest that the difference shown between the 111In and 67Ga conjugates might be mediated by differences in the in vivo chemistry of the metallic ions. PMID- 9228657 TI - 212Bi-DOTMP: an alpha particle emitting bone-seeking agent for targeted radiotherapy. AB - The synthesis and in vivo stability of the bone-seeking alpha-particle emitting compounds 212Bi-DOTMP and 212Pb/212Bi-DOTMP are described. 212Bi-DOTMP, injected i.v. into Balb/c mice, showed prominent bone localization and a rapid clearance from blood and other organs. Femur/blood ratios increased from 13 at 15 min up to 490 at 2.0 h postinjection. Enhanced uptake of 212Bi-DOTMP was demonstrated in regions with high bone turnover. A comparison between 212Bi-DOTMP and [153Sm]Sm EDTMP showed essentially no differences in biodistribution. 212Pb/212Bi-DOTMP followed a similar biodistribution, except for slightly elevated levels of 212Bi in the kidneys. The present study has shown 212Bi-DOTMP to be an in vivo stable bone-seeking radiopharmaceutical with promising biological properties for the treatment of sclerotic metastases and osteoblastic osteosarcoma. PMID- 9228658 TI - Retardation of 17-oxidation of 16 alpha-[18F]fluoroestradiol-17 beta by substitution of deuterium for hydrogen in the 17 alpha position(6). AB - We describe the synthesis, in vitro metabolism and biodistribution of [17 alpha 2H]16 alpha-[18F]fluoroestradiol ([18F]DFES). The clinically useful breast cancer imaging agent, 16 alpha-[18F]fluoroestradiol-17 beta ([18F]FES), was deuterated at the C-17 alpha position to lower the rate of C-17 alcohol oxidation. Metabolism studies in immature female rat and mature female baboon isolated hepatocytes showed [18F]DFES being consumed ca. 2.5 times slower than [18F]FES. Biodistribution studies and time-activity curve measurements in female rats showed [18F]DFES to have superior uptake characteristics compared to [18F]FES for imaging estrogen-receptor rich targets. PMID- 9228659 TI - Radiosynthesis and PET imaging of [N-methyl-11C]LY257327 as a tracer for 5-HT transporters. AB - No-carrier-added [N-methyl-11C]LY257327 was synthesized by methylation of the free base of the desmethyl precursor LY214281 with [11C]methyl iodide in anhydrous acetonitrile. Synthesis time was 52 +/- 3 min, radiochemical yield (based on [11C]methyl iodide) was 35 +/- 8%, radiochemical purity was 99 +/- 1%, and specific activity at EOB was 3900 +/- 1300 mCi/mumol. Two in vivo studies in baboon were carried out before and after pretreatment with the selective serotonin reuptake inhibitor citalopram. The first experiment showed high accumulation of radioactivity in midbrain, striatum, and thalamus, with slightly lower accumulation in the occipital and cerebellum regions. The radioactivity concentration peaked 5 min postinjection, decreasing steadily for the rest of the scanning time. The second experiment (blocked with citalopram) showed only partial inhibition of incorporation in all of the same brain regions. Although [N methyl-11C]LY257327 displayed high brain uptake (5% of injected dose at 5 min postinjection) and localized in serotonergic areas of the brain, its target-to nontarget ratio and its insensitivity to citalopram blocking suggest that its accumulation is dominated by nonspecific uptake. Therefore, [N-methyl 11C]LY257327 is not a useful agent for measuring serotonin reuptake sites in vivo by positron emission tomography. PMID- 9228660 TI - Tissue distribution and radiation dosimetry of astatine-211-labeled chimeric 81C6, an alpha-particle-emitting immunoconjugate. AB - A paired-label study was performed in athymic mice bearing subcutaneous D-54 MG human glioma xenografts to compare the localization of human/mouse anti-tenascin chimeric antibody 81C6 labeled by reaction with N-succinimidyl 3 [211At]astatobenzoate and N-succinimidyl 3-[131I]iodobenzoate. Over the 48-h observation period, the distribution of 211At- and 131I-labeled antibody were quite similar in tumor and normal tissues except stomach. These data were used to calculate human radiation doses for both intravenously and intrathecal administered 211At-labeled chimeric 81C6 using a quality factor of 5 for alpha emissions. PMID- 9228661 TI - Effect of endogenous biotin on the applications of streptavidin and biotin in mice. AB - The use of streptavidin-conjugated antibody to pretarget tumors in animals and patients, prior to administration of radiolabeled biotin, has provided encouraging results, in part because of the high affinity of biotin for streptavidin and the rapid whole-body clearance of biotin. However, binding of endogenous biotin to streptavidin may interfere with the clinical potential of this approach. This report evaluates the effect of endogenous biotin on an antibody-streptavidin conjugate in a mouse tumor model. Tumored nude mice were depleted of endogenous biotin by sequential intraperitoneal injections of streptavidin. The assay of serum biotin levels indicated less than 0.5 ng of biotin per mL of serum in treated mice versus 4 ng per mL in untreated animals. Flow cytometric analysis was used on single-cell suspensions of tumor from animals receiving streptavidin-conjugated IgG to detect the presence of the antibody on the cell membrane (with fluoroisothiocyanate-conjugated goat anti mouse antibody), and to detect biotin binding sites on streptavidin (with biotin phycoerythrin). Both treated and untreated mice demonstrated the presence of antibody on tumor cells through 48 h postadministration, but only in treated animals were biotin binding sites observed. These results in the mouse model suggest that the small concentration of streptavidin delivered to a tumor via a specific antibody may be saturated with endogenous biotin and therefore not able to be targeted subsequently with radiolabeled biotin. PMID- 9228662 TI - Application of a novel phenylpiperazine formation reaction to the radiosynthesis of a model fluorine-18-labeled radiopharmaceutical (18FTFMPP). AB - The labeled serotonin agonist 3-[18F]fluoro-N-(alpha,alpha,alpha-trifluoro-m tolyl)piperazine (18FTFMPP) was prepared rapidly using the labeling procedure for trifluorotoluenes, [18F]fluoro-for-nitro exchange, followed by an alumina supported bis-alkylation. After normal-phase HPLC purification, the labeled product was obtained in 20-32% (n = 20) decay-corrected radiochemical yield with a radiochemical purity > 98% and a specific activity of 100 GBq/mumol. The synthesis time including purification was 3 h. The receptor binding affinity of FTFMPP to rat brain membranes was found to be similar to that of the nonfluorinated parent compound (TFMPP). Although TFMPP has been proposed by others as an agent for the imaging of serotonin receptors, only minimal receptor mediated uptake was observed. PMID- 9228663 TI - International Union of Pharmacology Nomenclature in Nitric Oxide Research. PMID- 9228664 TI - The blood-brain barrier in neuroinflammatory diseases. PMID- 9228665 TI - Calcium movements, distribution, and functions in smooth muscle. PMID- 9228666 TI - Detection of loss of heterozygosity in the APC tumor suppressor gene in nonpapillary renal cell carcinoma by microdissection and polymerase chain reaction. AB - The role of the APC (adenomatous polyposis coli) tumor suppressor gene in the genesis of nonpapillary renal cell carcinoma is addressed. The frequency of allelic deletion in the APC gene was analyzed using microdissection of the tumor specimens and a PCR (polymerase chain reaction)-based assay for the detection of intragenic loss of heterozygosity (LOH). Twelve of 29 carcinomas investigated were informative (41%). In five of these (42%) LOH was detected in the APC gene, LOH did not correlate with tumor grade or stage. This high frequency of intragenic LOH suggests an implication of the APC gene or a closely linked gene in the genesis of a subset of nonpapillary renal cell carcinoma. The use of a microdissection technique allows the reliable detection of tumor-specific LOH when using a PCR-based assay. PMID- 9228667 TI - Tubular PAH transport capacity in human kidney tissue and in renal cell carcinoma: correlation with various clinical and morphological parameters of the tumor. AB - In vitro accumulation of p-aminohippurate (PAH) was investigated in "intact" human renal cortical slices of normal kidney tissue and in tissue slices of renal cell carcinoma (RCC). The technique used was established in preliminary experiments on rat kidney tissue slices. In principle, the accumulation capacity is comparable in renal tissue slices of both species (slice to medium accumulation ratios between 4 and 8). In man sex differences in accumulation capacity do not exist. But, as shown in detail for rats, accumulation capacity drops with age. Tissue slices of RCC are unable to accumulate PAH actively; slice to medium ratio reaches about 1 and indicates passive PAH uptake only. Surprisingly, in tumors of stage pT1 PAH uptake is lowest. perhaps as a sign of PAH transport out of the cells. There is no difference between peripheral and central parts of RCC. Age and sex are without influence on PAH uptake in RCC tissue slices. Interestingly, the accumulation capacity of "intact" tissue of kidneys infested with RCC also depends on the severity of the tumor (stage, diameter), but not on grading and formation of metastases. PMID- 9228668 TI - Elevated concentrations of the small stress protein HSP27 in rat renal tumors. AB - The expression of two small stress proteins, alpha B crystallin and the 27-kDa heat shock protein (HSP27), was studied quantitatively and immunohistochemically in normal kidney and renal tumors in rats. Levels of alpha B crystallin in renal cell tumors tended to be higher than in normal kidney (P = 0.07), but with a wide range of values, whereas they were significantly lower in mesenchymal tumors (P < 0.0001). In contrast, HSP27 concentrations in both renal cell (mean +/- SD: 1790 +/- 940 ng/mg protein, n = 15) and mesenchymal (1260 +/- 1080 ng/mg protein, n = 10) tumors were significantly higher than the normal kidney value (142 +/- 30 ng/mg protein, n = 10, P < 0.0001). A positive correlation was found between alpha B crystallin and HSP27 levels limited to the renal cell tumor case (Pearson's correlation coefficient, r = 0.68, P < 0.01). Immunohistochemistry revealed the loops of Henle to be positive for alpha B crystallin, whereas HSP27 staining was positive in glomerular and interstitial vascular walls and epithelial cells of proximal and distal tubules. Positive immunostaining for alpha B crystallin was demonstrated in six of nine renal cell tumors (67%) studied and for HSP27 in all of the nine cases (100%). PMID- 9228669 TI - A model of orthotopic murine bladder (MBT-2) tumor implants. AB - We produced a model of orthotopic transplantation in C3H/He mice by transplanting MBT-2 cells. A single-cell suspension of 1.0 x 10(5) MBT-2 cells/0.02 ml was successfully injected into the submucosal layer of the bladder, with an incidence of 100% after four experimental weeks. Inoculated tumor grew expansively into the bladder cavity from the bladder submucosa and invaded the serosa and the surrounding tissue. This model more closely resembled the characteristics of human bladder tumor when compared to other bladder cancer models. The results of the histological observation, electron microscopic examination and DNA content analysis by flow cytometry showed that the transplanted carcinoma maintained the biologic characteristics of the primary tumor. PMID- 9228670 TI - Endoscopic observation for detection and monitoring of N-butyl-N-(4 hydroxybutyl)nitrosamine--induced bladder tumor in rats. AB - Recent advances in tumor carbohydrate biochemistry have demonstrated antitumor effects of locally administered GM3 ganglioside on mouse MBT-2 tumor. When intravesical therapy in N-butyl-N(4-hydroxybutyl)nitrosamine (BBN)-induced rat bladder tumor is attempted, it is essential to identify the tumor, to classify its size before therapy and to monitor the effect of the therapy. To establish a more reliable experimental therapeutic system, we assessed the development of BBN induced rat-bladder tumor by endoscopic observation. BBN-induced bladder tumors in rats were observed serially using a 4.2-F flexible fiberscope. The endoscopic findings were compared with the histopathological findings. Intravesical tumor growth varied greatly between individual rats. The smallest change detected by endoscopy was a small edematous lesion histologically proved to be papilloma. The largest nodular lesion was determined to be a papillary, transitional cell carcinoma. This noninvasive method makes the BBN rat experimental system more reliable by allowing confirmation of tumor formation and classification of the tumor volume prior to therapy. PMID- 9228671 TI - Expression of CD44V2 in transitional cell carcinoma of the urinary bladder and in urine. AB - CD44 is the principal cell surface receptor for hyaluronate. Variant forms of the receptor, produced by alternative splicing, have been found to be associated with tumor progression in a variety of cancers. Based on investigations at the RNA level, it has recently been proposed that expression of CD44 variant V2 was present in urothelial cancer but not in normal urothelium. Since a distinctive marker for urothelial cancer would be extremely useful, frozen sections of normal urothelium and urothelial cancer were examined for expression of standard CD44 and CD44V2. Frozen sections of specimens of 35 patients with transitional cell carcinoma of the bladder, 16 specimens of normal bladder and 5 ureters were examined. Immunohistochemical staining was performed using a polyclonal antibody to CD44V2 (PAB CD44V2), a monoclonal antibody to CD44V2 (MAB CD44V2) and a monoclonal antibody to CD44S (MAB CD44S). CD44V2 and CD44S were also measured in lysates of urine sediments from 21 patients by enzyme-linked immunoabsorbent assay (ELISA). All investigated transitional cell carcinomas expressed CD44V2. There was no differentiation between invasive and noninvasive carcinoma. CD44V2 was also expressed in normal urothelium. Standard CD44 was expressed by the transitional cell carcinoma, normal urothelium, musculature and interstitial tissue. The amount of CD44V2 and CD44S in lysates of urine sediments is not correlated to diagnosis. In contrast to investigations at the RNA level, CD44V2 on the protein level seems not to be a distinctive marker for urothelial cancer. Therefore, CD44V2 will not be a useful diagnostic marker for detection of transitional cell carcinoma. PMID- 9228672 TI - Restoration of rat bladder function following release of short- and long-term partial outflow obstruction. AB - Detrusor dysfunction does not recover in some patients with benign prostatic hyperplasia (BPH) even after prostate resection. 'We studied the functional restoration of the rat bladder after release of short- or long-term outflow obstruction. Bladder function was assessed by in vivo infusion cystometry and an in vitro organ bath technique. There were no significant differences in bladder weight and contractile strength induced by stimuli in detrusor muscle strips from obstructed rats and age-matched control rats. After short-term obstruction the whole bladder pressure generated in vitro by field stimulation, bethanechol, ATP, and KCl completely recovered to control levels. In contrast, after long-term obstruction, the whole bladder pressure in response to field stimulation remained significantly lower than in controls. Infusion cystometry variables, including the pressure at which micturition was induced, maximal voiding pressure, capacity, and residual urine volume, were similar between controls and rats subjected to short-term obstruction. However, the maximal voiding pressure after long-term obstruction was significantly less than that of controls. PMID- 9228673 TI - Comparison of adrenoceptor subtype expression in porcine and human bladder and prostate. AB - We have quantified and characterized alpha 1-, alpha 2- and beta-adrenoceptor subtypes in porcine bladder detrusor and bladder neck, human bladder detrusor, and porcine and human prostate. alpha 1-, alpha 2- and beta-adrenoceptor were identified in radioligand binding studies using [3H]prazosin, [3H]RX 821002 and [125I]iodocyanopindolol, respectively, as the radioligands. In porcine male and female detrusor and bladder neck and male prostate, adrenoceptors were detected in the order of abundance beta > alpha 2 >> alpha 1 (not detectable), with no major difference between the sexes or between detrusor and bladder neck. In human detrusor and prostate the order of abundance was beta > alpha 2 >> alpha 1 (not detectable) and beta >> alpha 1 > alpha 2, respectively. The alpha 2 adrenoceptors in all tissues were homogeneously of the alpha 2A-subtype as evidenced by competition binding studies with yohimbine, prazosin, ARC 239 and oxymetazoline. The beta-adrenoceptors represented a mixed population with a dominance of the beta 2-subtype in all tissues as demonstrated by competition binding with ICI 118,551 and CGP 20,712A. We conclude that pigs may be a suitable model for studies of detrusor function with respect to adrenoceptor expression. They may be less suitable for studies of bladder neck or prostate function. PMID- 9228674 TI - Effect of ischemia and partial outflow obstruction on rat bladder function. AB - We investigated the effects of ischemia induced by ligation of the bilateral internal iliac arteries following partial outlet obstruction on changes in detrusor function in rat. Rats were divided into three groups: sham-operated control rats, rats with partial outlet obstruction, and rats with obstruction+ischemia. Bladder function was studied by the in vitro organ bath technique 7 days after surgery. The weight of the bladder was significantly increased in both the obstruction and obstruction+ischemia groups. The obstruction+ischemia group exhibited a greater increase in weight. The passive length-tension relationship of detrusor muscle strips showed that tissue elasticity was decreased and the active length-tension relationship demonstrated that the peak response was observed at a shorter tissue length in the obstruction+ischemia group compared with the other two groups. There was no difference in the passive and active length-tension relationships between the control group and the obstruction group. The contractile response to various kinds of stimulation (field stimulation, bethanechol, ATP, and KCl) increased in the obstruction group and decreased in the obstruction+ischemia group. These findings suggest that partial outflow obstruction alone increased bladder contractility in response to stimuli. However, ischemia reduced the contractility and elasticity of the bladder wall. PMID- 9228675 TI - Is it possible to prevent bacterial adhesion onto ureteric stents? AB - The aim of this study was to determine whether the use of bactericidal coatings or immersion in antibiotic solution reduces or prevents bacterial adhesion onto ureteric stents. Precut segments of full silicone, silver-coated and hydrogel coated ureteric stents were incubated with two uropathogenic bacterial strains with and without previous immersion in antibiotic solution. Tobramycin, ceftriaxone and ciprofloxacin solutions were used, as these antibiotics are commonly administered for the prophylaxis and treatment of urinary tract infection (UTI). Microbiological analysis showed that immersion of ureteric stents in ceftriaxone and ciprofloxacin yielded a significant reduction of bacterial adhesion, whereas immersion in tobramycin did not. The surface material of the stents had no direct influence on bacterial adhesion. In this experimental study, neither the silver nor the hydrogel coat reduced bacterial adhesion onto ureteric stents whereas immersion in a suitable antibiotic solution significantly reduced and even prevented this phenomenon, probably due to the adhesion of the antibiotic onto the stent surface. Prevention of bacterial adhesion onto ureteric stents is essential to reduce the risk of UTI in connection with these devices. PMID- 9228676 TI - Urine lipids in patients with a history of filariasis. AB - The presence of lipids in postprandial urine was assessed in 116 patients with a history of filariasis and 70 normal individuals using a biochemical autoanalyzer. Urinary triglycerides (TGs) ranging from 10 to 1955 mg/dl were detected in 13 individuals with a history of chyluria, including 3 with TG levels ranging from 233 to 1955 mg/dl and cholesterol levels of 6-35 mg/dl. Three patients who had a filarial history but without chyluria were also found to have urinary TGs (13-15 mg/dl) without detectable cholesterol. Neither TGs nor cholesterol were detected in the urine of normal individuals. Fluctuations in postprandial urine lipid contents were measured by time course determinations of urinary TG and cholesterol in 17 patients with filariasis and a history of chyluria, 16 patients with filariasis and hydrocele and 16 normal individuals. The level of urine lipid excretion was found to increase within 1-4 h postprandially, with urinary TG levels ranging between 7.8 and 1284 mg/h in eight patients and urinary cholesterol levels between 1.2 and 138 mg/h in seven patients with a history of chyluria. To evaluate the origin of the urine lipids in hematochyluria, fish oil containing 360 mg eicosapentaenoic acid (EPA) and 240 mg docosahexaenoic acid (DHA) was prescribed to a patient with hematochyluria. The excretion of EPA and DHA in urine was increased postprandially in the patient, similar to the elevation of urinary TG and cholesterol. The profile of fatty acids from urine samples showed it was of dietary origin. Our results suggest that postprandial urine lipids, especially TG, might be used as markers for the clinical evaluation of chyluria. PMID- 9228677 TI - Evaluation of a blocking Elisa for screening of antibodies against porcine reproductive and respiratory syndrome (PRRS) virus. AB - A blocking Elisa was developed for the detection of antibodies against PRRS virus with a view to satisfying the need for examination of blood samples on a large scale. The test was evaluated in comparison with an indirect Elisa and the immunoperoxidase monolayer assay. The blocking Elisa was sensitive and specific. It had a higher capacity and was cheaper to perform than the immunoperoxidase monolayer assay and the indirect Elisa. It was comparable to the immunoperoxidase monolayer assay and better than the indirect Elisa in detecting antibodies formed early after infection, and it was superior to both the immunoperoxidase monolayer assay and the indirect Elisa in detecting antibodies at a late stage of infection. PMID- 9228678 TI - Virus quantification and identification of cellular targets in the lungs and lymphoid tissues of pigs at different time intervals after inoculation with porcine reproductive and respiratory syndrome virus (PRRSV). AB - Sixteen 6 week old conventional pigs were inoculated by aerosol with a European strain of porcine reproductive and respiratory syndrome virus (PRRSV). Virus replication was followed by virus titration and immunofluorescence in the lungs and in associated and distant lymphoid tissues at 3, 14, 21, 35, 42 and 82 days post-inoculation (DPI). PRRSV replication was detected in alveolar macrophages, lungs, tonsils, spleen, retropharyngeal lymph nodes, bronchial lymph nodes and thoracic aortic lymph nodes at 3 DPI. The same tissues, except retropharyngeal and thoracic aortic lymph nodes, were PRRSV positive at 14 DPI. Lungs and alveolar macrophages were PRRSV positive until 35 DPI. PRRSV was not detected in heart, peripheral blood mononuclear cells and bone marrow cells. Viremia was detected from 3 to 28 DPI. Not more than 2% of alveolar macrophages were PRRSV positive even during the acute stage of infection. 80 to 94% of the PRRSV infected cells in the lungs and in lung lavaged cells were identified as macrophages using a porcine macrophage specific monoclonal antibodies. In the lymph nodes and spleen, 100% of the infected cells were macrophages. Anti-PRRSV antibodies were detected by a blocking ELISA as early as 7 DPI. the antibody titre gradually increased to reach a geometric mean titre (GMT) of 160 at 35 DPI. It remained at that level until the end of the study. These findings clearly demonstrate that PRRSV has a tropism for macrophages. PRRSV mainly replicates in macrophages of the lymphoid tissues and lungs in the acute phase of infection and persists in the lung macrophages. PMID- 9228679 TI - Seroprevalence of porcine reproductive and respiratory syndrome virus in Dutch weaning pigs. AB - To determine whether under Dutch field conditions PRRSV infection occurs in weaning pigs before the finishing period, a cross-sectional study was performed on 32 breeding farms to estimate the seroprevalence of antibodies directed against PRRSV in 4- to 5-week-old and 8- to 9-week-old pigs. Farms were visited twice within 5 months, and during each sampling an average of 20 sera were randomly collected from a unit of 4- to 5-week-old and a unit of 8- to 9-week-old pigs. The sera (n = 2568) were tested in the IDEXX-ELISA for the presence of antibodies directed against PRRSV. The seroprevalence of PRRSV in 4- to 5-week old pigs and 8- to 9-week-old pigs varied between both samplings for each farm. The seroprevalence in the younger pigs was significantly higher than in the older pigs for both samplings (p < 0.05), suggesting the presence of maternal antibodies. In addition, a longitudinal study was performed to evaluate the IDEXX ELISA in detecting maternal antibodies directed against PRRSV and to determine the rate of decline of these antibodies in field sera. From serological results of eight litters, an average decay function was computed to quantify the maternal immunity to PRRSV. A seroprevalence in 8- to 9-weeks-old pigs of > or = 0.20 was calculated to indicate an active immune response to PRRSV. In the cross-sectional study in the pigs twenty-three percent of the units with 8- to 9-week-old pigs were considered to have an active serological response against PRRSV. We conclude that most Dutch pigs are seronegative for PRRSV at the start of the finishing period, since the results of this study showed that 77% of the units with 8- to 9 week-old pigs had a seroprevalence < 0.20. PMID- 9228680 TI - Comparison of the primary signs induced by experimental exposure to either a pneumotrophic or a 'limping' strain of feline calicivirus. AB - Two strains of feline calicivirus, one reportedly pneumotrophic (FPV 255) and the other associated with a limping syndrome (2280) were compared with respect to the signs induced in kittens after oronasal exposure. Neither strain induced severe upper respiratory symptoms, and both caused oral ulcers and lameness. However oral ulcers were more prevalent, and lameness and depression were more pronounced in the kittens which received strain 2280. Kittens which exhibited lameness also had elevated blood levels of alpha 1-acid glycoprotein. A decline in lymphocyte count was noted only in kittens which received strain 2280. These data demonstrate that despite reported antigenic and genetic differences between these strains, no distinct differences in pathogenicity could be determined. PMID- 9228681 TI - Attenuation of dUTPase-deficient pseudorabies virus for the natural host. AB - Pseudorabies virus (PrV) is the causative agent of Aujeszky's disease which results in significant losses in pig husbandry. Recently we identified the gene encoding the deoxyuridine-triphosphatase (dUTPase) of PrV as the homolog of the UL50 gene of herpes simplex virus type 1. The PrV UL50 gene product was characterized and a UL50 negative PrV mutant (PrV UL50-) was generated by insertion of a lacZ expression cassette into the UL50 open reading frame (Jons and Mettenleiter, J. Virol. 70, 1242-1245). Here we show that replication of PrV UL50- in cell culture was only slightly impaired as compared to wild-type PrV strain Ka. After intranasal infection of young pigs PrV UL50- proved to be substantially attenuated, whereas severe clinical signs and death occurred after infection with wild-type PrV. Challenge infection with the highly virulent NIA-3 strain of PrV showed that prior infection with PrV UL50- conferred protection against Aujeszky's disease. Innocuity and efficacy make UL50-negative PrV an attractive candidate for a live PrV vaccine. PMID- 9228682 TI - Serological analysis of feline calicivirus isolates from the United States and United Kingdom. AB - The antigenic relationship of 181 feline calicivirus (FCV) isolates from the United States (USA) and United Kingdom (UK) to 5 reference viruses, representing 2 vaccine strains (F9 VacNor, 2280) and 3 field strains (H, J, TN) was determined by a neutralization assay using two standardized dilutions of reference sera. A comparison between USA and UK isolates indicated that significantly fewer UK isolates could be neutralized by sera specific for each of the reference strains. Isolates from the USA were also analyzed based on the time period and geographical location from which they were derived. Analysis of the cross neutralization results for isolates derived from the USA indicated that no single reference strain was predominant in all the geographical locations, however, antisera specific to strains 2280, H and J neutralized the highest number of isolates overall. Analysis of the neutralization patterns based on the time period from which the isolates were derived indicated that F9 VacNor was antigenically similar to 86% of feline caliciviruses isolated in 1958-1979. This number decreased to 43% for isolates derived in 1980-1995. By contrast, the J strain maintained its antigenic relatedness to approximately 75% of representative FCV isolates circulating during the last 40 years. PMID- 9228683 TI - Flagellin, a major protein present in SDS-PAGE profiles of Sarkosyl-OMP-enriched fractions from Bordetella bronchiseptica Bvg- or modulated Bvg+ strains. AB - The bvg or vir locus positively regulates the expression of many Bordetella virulence-associated determinants (encoded by vag genes), including cell envelope proteins, in response to environmental stimuli. On the other hand, several genes named vrg genes are negatively controlled by the bvg regulon (Knapp and Mekalanos, 1988). Flagellin is encoded by a vrg gene, which is expressed when the principal virulence factors are eliminated during antigenic modulation or in phase variants (Akerley et al., 1992). We have previously analyzed SDS-PAGE profiles of Sarkosyl-outer membrane protein (OMP)-enriched fractions from B. bronchiseptica Bvg- and modulated Bvg+ strains and reported a major band associated with the avirulent phenotype (Passerini de Rossi et al., 1995). In order to characterize this band we have purified flagellar filaments from Bvg- and modulated Bvg+ strains, and analyzed them by SDS-PAGE. These profiles revealed a single major band of 40 or 45 kDa depending on the strain. The N terminal amino acid sequence of the putative flagellin expressed by BB7200a was identical over the first 21 residues analyzed to that of the flagellin from the modulated strain BB7865 reported by Akerley et al. (1992). Comparison of the SDS PAGE profile of flagellar filaments with that of the OMP-enriched fraction of the corresponding strain showed that the flagellum-associated polypeptide had the same electrophoretic mobility as that of the characteristic band of the avirulent phenotype. Furthermore, this band was absent in the OMP-enriched fraction profile from a Bvg- strain subjected to a treatment that removes flagella. Our results indicate that the major protein observed in SDS-PAGE profiles of Sarkosyl-OMP enriched fractions from B. bronchiseptica Bvg- and modulated Bvg+ strains corresponds to flagellin. PMID- 9228684 TI - Comparison of an LPS-specific competitive ELISA with a motility enrichment culture method (MSRV) for detection of Salmonella typhimurium and S. enteritidis in chickens. AB - We report here evaluation of a competitive enzyme-linked immunosorbent assay (c ELISA) for detection of Salmonella spp. in chicken organs and faeces. The c-ELISA used a monoclonal antibody (MAb), specific for a genus-specific epitope of the outer core oligosaccharide of salmonellae. Salmonella lipopolysaccharide (LPS) in samples competed with Salmonella LPS coated on microtitre plates, for binding to the MAb. Competition reduced binding of the MAb to the LPS on the plate and of the secondary antibody to the MAb hence reducing the chromogenic signal. Stable coating and minimal false positive were achieved by conjugating LPS to poly-L lysine. The c-ELISA was compared with motility enrichment culture using modified semisolid Rappaport Vassiliadis (MSRV) medium, which detected less than 10(2) CFU/g, and did not allow migration of non-salmonella species. The c-ELISA detected 10(6) CFU of enriched culture or 10(2)-10(3) CFU of Salmonella/g of faeces. Its limit of detection was thus higher than that of MSRV culture and it had a sensitivity of 92.9% and a specificity of 96.7%. PMID- 9228685 TI - Development of a specific DNA probe and PCR for the detection of Mycoplasma bovis. AB - Mycoplasma bovis is responsible for several production diseases in cattle, including mastitis, arthritis, pneumonia, abortion and infertility. Current methodologies for detecting and identifying M. bovis are time consuming and difficult. Tests which rely on antigen or antibody detection have poor sensitivity and specificity. In this paper associated protocols for the development of a hybridization probe and PCR are described. A genomic library (SauIIIA digested) was prepared from M. bovis DNA (Colindale Reference Strain: NC10131:02) and cloned into pUC19. Colony hybridization, using a probe preparation made from purified M. bovis DNA, was used to identify colonies of interest. M. bovis DNA fragments were retrieved from recombinant plasmids by digestion with EcoRI and HindIII. This DNA was used to prepare randomly primed probes for dot blot hybridization analysis with immobilized DNA from M. bovis (two strains), M. dispar, M. agalactiae, M. bovigenitalium (two strains), M. ovipneumoniae, a Group 7 strain, M. arginini and bacteria belonging to different genera. Four probes were found to hybridize only with M. bovis and M. ovipneumoniae DNA, whereas one probe reacted with genomic DNA from only one of the two M. bovis strains. The level of sensitivity of the dot blot hybridization assay was 200 CFU (colony forming units)/mL. To enhance the sensitivity further, an M. bovis-specific PCR assay was developed. The primers were designed using sequences obtained from the probe DNA which discriminated M. bovis from all other Mycoplasma DNA tested. The minimum amount of target DNA that could be detected by the PCR assay was that isolated from 10-20 CFU/mL. The PCR assay was therefore 10 times more sensitive than dot blot hybridization. PMID- 9228686 TI - Pathogenesis of Salmonella enteritidis phage type four after experimental infection of young chickens. AB - White leghorn specific-pathogen-free chickens were inoculated orally with Salmonella enteritidis phage type 4 at the age of one day (group 1) and four weeks (group 2). From 3 h until 4 weeks post inoculation (pi), birds were sacrificed. Gross lesions were recorded and different sites of the intestine and visceral organs were collected for bacteriological and histopathological examination. Clinical disease and mortality were only observed in group 1. Mortality was 8%. The birds were depressed, had diarrhoea and an indurated yolk sac. Infection of the liver and the heart was present within 12 h pi in both groups. The percentage of infected organs was very high and similar in both groups during the first week pi. Thereafter the isolation rate of Salmonella was declining faster in group 2. The crop, the proventriculus, the lower intestinal tract and the bursa of Fabricius were the predilective sites of isolation in both groups. Most prevalent lesions were serous typhlitis, omphalitis and polyserositis. Histopathology revealed inflammation in the intestines and visceral organs. In some birds granulomatous nodules were present in the caeca. Antibodies were detected from 18 and 5 days pi in group 1 and 2, respectively. Granulomatous nodules were typical of infection with this strain of S. enteritidis phage type 4. These granulomatous nodules together with the retained yolk sac possibly are a source of Salmonella organisms that may account for intermittent faecal shedding by carrier birds. PMID- 9228687 TI - Epidemiology and virulence assessment of Salmonella dublin. AB - Six strains of Salmonella dublin with distinct antimicrobial susceptibility patterns and/or plasmid profiles were repeatedly isolated from calves in a calf rearing facility. Three of the six strains were isolated from numerous calves during outbreaks of clinical salmonellosis while the other three were not. These strains were compared for their ability to adhere to and internalize in human intestinal epithelial cells (Caco-2) and in bovine alveolar macrophages (BAM), to survive in BAM, and to cause lethal infection in female BALB/c mice. All six strains of S. dublin demonstrated an ability to adhere to and internalize in both Caco-2 cells and in BAM. However, strain differences in the level of adhesion and/or internalization in Caco-2 cells and BAM were demonstrated. Most strains were able to persist but not proliferate in BAM. One outbreak-associated strain which readily attached and internalized in eukaryotic cells in vitro was avirulent to mice at the dose tested. The remaining five strains were virulent to mice. In vitro measures of virulence attributes were not clearly correlated with virulence among S. dublin strains measured either as prevalence in calves during outbreaks of disease or as mouse lethality. Also, there was no association between prevalence of strains in calves during outbreaks of clinical salmonellosis and lethality in mice. PMID- 9228688 TI - Analysis of Haemophilus parasuis by multilocus enzyme electrophoresis. AB - The diversity among 40 Australian isolates and eight reference strains of Haemophilus parasuis was examined using multilocus enzyme electrophoresis. Thirty four electrophoretic types (ETs) were recognized with a mean diversity per locus of 0.405. One Australian isolate was located in an ET separated by a considerable distance (> 0.8) from the rest of the isolates, suggesting that it may represent a different species or subspecies. The remaining 33 ETs formed two distinct divisions (A and B), separated from each other by a distance of 0.506. All 12 Australian isolates of serovar 5 plus the two reference strains for this serovar were included in Division A. The only other isolates present in this Division were Australian isolates of serovars 4 and 13 and two nontypeable isolates. Division B contained a diverse range of serovars-Australian isolates of serovars 1, 2, 7/10, 9 and 13 as well as the reference strains for serovars 1, 2, 3, 4, 8 and 9. These results supported other studies which demonstrated considerable diversity, and indicate that the population of H. parasuis may contain more than a single species or subspecies. There was considerable diversity even amongst isolates of the same serovar, indicating that serotyping is not a particularly suitable technique for strain typing in epidemiological studies. PMID- 9228689 TI - Magnetic immuno capture PCR assay (MIPA): detection of Leptospira borgpetersenii serovar hardjo. AB - Magnetic immuno PCR assay (MIPA) was developed for the rapid detection of leptospires excreted in urine samples (n = 59) collected from 35 experimentally infected cattle. The immunomagnetic separation of leptospires from inhibitors in frozen formalin fixed bovine urine prior to PCR detection resulted in a marked improvement on previous detection methods. MIPA is a rapid 5 step protocol requiring 70 mins preparation time prior to amplification, which consistently detects 10(1) organisms. MIPA detected 76% (38/50) of culture positive urines and in addition three urines that were culture negative were shown to be positive by this method of detection. Consequently we conclude that whilst MIPA is an improvement on previously published PCR detection methods, the culture of the organism is still the standard against which other detection methods have to be compared. PMID- 9228690 TI - Molecular and phenotypic methods for the characterization of atypical Aeromonas salmonicida. AB - Atypical Aeromonas salmonicida form a taxonomically diverse group among the psychrophilic A. salmonicida. Characteristics of 53 atypical A. salmonicida strains originating from Finland, Denmark, Norway and Sweden were studied using 60 phenotypic tests. Ribopattern analysis and plasmid profiles were used as genetic methods. The production of brown pigment on the furunculosis agar containing L-tyrosine divided the atypical oxidase-positive strains into two groups: pigment-producing (n = 35) and achromogenic (n = 16). PstI differentiated all the atypical A. salmonicida from A. salmonicida subsp. salmonicida. Combined ribopatterns ClaI/PstI divided pigment producing atypical strains into four major groups B/B, G/G, G/T, F/F. Most of the achromogenic oxidase-positive strains belonged to two major groups H/H or U/U. Cluster analysis of ribopatterns and plasmid profile analysis also supported the division of atypical oxidase-positive A. salmonicida into pigment-producing and achromogenic groups. The oxidase negative strains formed a distinct group which differed from oxidase-positive atypical A. salmonicida type biochemically and in terms of ribopatterns. Our results also support the use of ribopattern analysis as a valid method to study the epidemiology of infections caused by atypical A. salmonicida on fish farms. PMID- 9228691 TI - Influence of orchiectomy on canine behaviour. AB - One hundred and twenty-two dog owners were interviewed to obtain information about the effects of orchiectomy on the behaviour, unwanted side effects, and testosterone-dependent disease processes in their dogs. Behavioural problems were the main reason for orchiectomy, unwanted sexual behaviour being the most common, together with roaming, aggression, and abnormal urination behaviour. Objectionable sexual behaviour, inter-male aggression, roaming, and abnormalurination were reduced after orchiectomy in approximately 60 per cent of the dogs. The side effects of orchiectomy included increased bodyweight, increased appetite and decreased activity in less than 50 per cent of the dogs, and there was a significant relationship between increased appetite and bodyweight. The clinical signs of testosterone-dependent disease in most of the dogs either decreased or disappeared after orchiectomy. PMID- 9228692 TI - Histological and bacteriological evaluation of digital dermatitis in cattle, with special reference to spirochaetes and Campylobacter faecalis. AB - Tissue samples from the feet of slaughtered cattle exhibiting different stages of digital dermatitis were sectioned and stained with haematoxylin and eosin and silver staining techniques. Three morphological variations of spirochaetes were observed, whereas control samples from feet which were macroscopically negative for digital dermatitis were also negative for spirochaetes. In an immunofluorescence test, Campylobacter faecalis was found to be abundant on superficial wound smears from the classical ulceration of digital dermatitis. PMID- 9228693 TI - Monensin toxicity in a flock of ostriches. AB - A total of 42 birds from a flock of 104 farmed ostriches showed signs of toxicity after the accidental inclusion of monensin in their concentrate ration. The initial clinical signs were muscle weakness and ataxia which progressed to recumbency, dyspnoea and death, despite intensive supportive therapy. The serum activity of the enzymes creatine kinase, aspartate aminotransferase and lactate dehydrogenase was high in the affected birds, indicating significant muscle pathology. Few gross lesions were identifiable postmortem, but widespread lesions of degenerative myopathy were present at the histopathological level. However, these degenerative changes were restricted to the skeletal muscle and there was no evidence of cardiomyopathy in any of the birds examined. The birds were fed a ration which contained 215 to 224 ppm monensin for 13 days. New clinical cases ceased to occur shortly after the withdrawal of the source of monensin, but all the individuals which showed clinical signs of toxicity died or were euthanased on humane grounds. PMID- 9228694 TI - Megabacteria in diseased and healthy budgerigars. PMID- 9228695 TI - Serum creatine kinase-MB and lactate dehydrogenase-1 isoenzyme activities in piglets with encephalomyocarditis virus. PMID- 9228696 TI - Persistent vitellointestinal duct in a calf. PMID- 9228697 TI - Bovine ephemeral fever in Saudi Arabia. PMID- 9228698 TI - Outbreak of staphylococcal dermatitis in housed lactating Suffolk ewes. PMID- 9228699 TI - Presentation of clean livestock for slaughter. PMID- 9228700 TI - Biosecurity systems and animal health. PMID- 9228701 TI - Caseous lymphadenitis: an increasing cause for concern. PMID- 9228702 TI - Caseous lymphadenitis: an increasing cause for concern. PMID- 9228703 TI - Integration of women into the profession. PMID- 9228704 TI - Canine angiostrongylosis in south Wales. PMID- 9228705 TI - Unusual cause of rumen stasis in a cow. PMID- 9228706 TI - Use of the Internet for research. PMID- 9228707 TI - Sex reversal in birds. PMID- 9228708 TI - Report of EPFA/NIBSC workshop 'nucleic acid amplification tests (NAT) for the detection of blood-borne viruses' held on 31 October 1996 in Amsterdam, The Netherlands. AB - The 3rd annual European Plasma Fractionators Association/National Institut of Biological Standard and Control (EPFA/NIBSC) meeting provided a forum for regulators, blood product and test kit manufacturers and organisations developing standards to present and discuss their latest data. The main conclusions were as follows. There has been substantial progress during the last year in the development of NAT technology specifically for improving the safety of blood products though there is an urgent need for the development of international reference materials. The technology is not yet sufficiently developed to be used as a routine screening test though testing of plasma pools for hepatitis C virus may be achieved within a year. Introduction of testing should not result in the creation of dual standards for plasma derived and cellular products. Once the technology is fully developed it could significantly improve the safety of all blood products, particularly those derived from starting materials with a high incidence of viral markers. PMID- 9228709 TI - Risk factors and anti-HBc reactivity among first time blood donors. AB - BACKGROUND AND OBJECTIVES: The usefulness of testing for antibody to hepatitis B core antigen (anti-HBc) as a surrogate marker for non-A, non-B hepatitis can no longer be clearly established in the face of anti-hepatitis C virus testing. Application of anti-HBc testing in blood donors for detection of hepatitis B in addition to hepatitis B surface antigen testing (HbsAg) is a matter of debate. MATERIALS AND METHODS: We examined the serology and risk analysis data in a group of first-time blood donors. In 1.48% of 16,081 donors, anti-HBc reactivity was found. We invited a study group of 112 donors for extensive interviewing about the risk of blood transmissible diseases, and for serological testing. A control group of 240 first-time donors was studied as well. RESULTS: In the study group, the age was older (p < 0.001), a history of liver disease was more frequent (p < 0.001), and the donor (p < 0.001) or the donor's partner (p < 0.05) had either stayed longer in an HBV-endemic area or had been born in one. Combining these with the serological results, we found that strong anti-HBc reactivity was related to hepatitis B risk factors in HBsAg-negative donors. CONCLUSIONS: Anti HBc testing in HbsAg-negative first-time donors makes it possible to identify hepatitis B risk factors with a prevalence of 0.02%. Our findings also stress the importance of including the history of the donor's partner(s) in the risk analysis before blood donation. PMID- 9228710 TI - Photosensitized inactivation of Plasmodium falciparum- and Babesia divergens infected erythrocytes in whole blood by lipophilic pheophorbide derivatives. AB - BACKGROUND AND OBJECTIVES: Blood transfusions can transmit parasitic infections, such as those caused by Plasmodium (malaria), Trypanosoma cruzi (Chagas' disease), and Babesia (babesiosis). A higher degree of blood transfusion safety would be reached if methods were available for inactivating such parasites. MATERIALS AND METHODS: We evaluated the effectiveness of photosensitization using lipophilic pheophorbide and red light illumination to eradicate red blood cells infected with Plasmodium falciparum, and with Babesia divergens, in whole blood. Fluorescence microscopy and conventional fluorometry showed the specific accumulation of pheophorbide derivatives in the RBC infected with either parasite, compared with uninfected RBC. The effectiveness of different derivatives in eradicating infected RBC was first estimated in parasite cultures. RESULTS: The best photosensitizer was the N-(4-butanol) pheophorbide derivative (Ph4-OH) at 0.2 microM concentration and 5-min illumination. In whole blood, the eradication of RBC infected with B. divergens and P. falciparum was obtained with 2 microM Ph4-OH and 10 and 20 min illumination, respectively. Under these conditions of photosensitization, low levels of RBC hemolysis were noted even after 2 weeks of storage at 4 degrees C and a subsequent 48-hour incubation at 37 degrees C. No reduction of negative charges on treated RBC was noted and no increase in methemoglobin content. CONCLUSIONS: In plasma, Ph4-OH is mainly transported by high-density lipoproteins (HDL). This high affinity for HDL may explain the selective accumulation of lipophilic pheophorbide derivatives in the intracellular parasites. Photosensitization with pheophorbide derivatives may be a promising approach to inactivation of transfusion-transmissible parasites and viruses in blood bank units. PMID- 9228711 TI - Effectiveness of leukocyte filters in reducing tumor cell contamination after intraoperative blood salvage in lung cancer patients. AB - OBJECTIVES: In an effort to reduce allogeneic blood transfusions in patients undergoing elective surgery for lung cancer, we investigated the effectiveness of a method of processing shed blood with an automated device for intra operative blood salvage (IOBS) and filtration with a 3rd-generation polyester filter to remove tumor cells. METHODS: Sixteen patients were operated on for different types of lung cancer. We searched for malignant cells in pre- and postprocessed shed blood employing density gradient centrifugation, staining of cytospins with hematoxylin-eosin, and antibodies to human cytokeratins. RESULTS: In 9 out of 16 cases (56%), neoplastic cells were detected in prefiltration samples, but none were found in postfiltration cytospins. CONCLUSION: IOBS combined with appropriate filtration could be a very useful and safe tool in reducing allogeneic blood transfusion in cancer patients. PMID- 9228712 TI - Prolonged holding of whole blood at 22 degrees C does not increase activation in platelet concentrates. AB - OBJECTIVES: Platelets prepared after holding of whole blood overnight at 22 degrees C have a well-preserved metabolism. However, the possibility that such prolonged incubation with active granulocytes may increase platelet activation has not been fully tested. METHODS: We investigated this possibility by flow cytometric analysis of membrane glycoproteins (GPs) Ib and IIb/IIIa and the activation markers CD62P and CD63 in platelet concentrates (PCs) prepared from whole blood that was held for either 6 h without cooling plates (n = 20) or for 24 h on cooling plates of 1,4-butanediol (n = 20). PCs were prepared by the platelet-rich plasma method and analyzed on the second storage day. RESULTS: Platelet yield and aggregation response to ristocetin, collagen and epinephrine + ADP were similar in both types of PCs, as was the mean fluorescence intensity for GPs Ib and IIb/IIIa. PCs prepared by the overnight-hold method did not differ from those obtained 6 h after collection in the percentage of platelets expressing CD62P (12.3 +/- 6.2% vs. 14.1 +/- 4.0%; p > 0.1) or CD63 (9.8 +/- 6.4% vs. 8.8 +/- 3.6%; p > 0.1). CONCLUSION: Prolonged holding of whole blood at 22 degrees C prior to component preparation does not increase the level of platelet activation. PMID- 9228713 TI - Citrate-dependent auto-antibody causing error in blood grouping. AB - BACKGROUND AND OBJECTIVES: Errors in blood grouping are occasionally due to anomalous antibodies or extraneous materials in the reagents. We investigated the case of a volunteer blood donor who was originally grouped as AB, Rh-positive, by slide grouping. However, her washed red cells showed group B, which agreed with her reverse serum grouping. MATERIALS AND METHODS: Standard serologic techniques were employed throughout. RESULTS: Investigation revealed an unusual autoantibody in the donor's blood that preferentially agglutinated the red cells in the presence of citrate. Because the grouping reagents contained citrate, this led to a false grouping when whole blood was tested rather than washed red cells. The antibody reacted at 37 degrees C and at 4 degrees C, but failed to agglutinate red cells when 0.1 M solution of monosaccharides was added to the reaction mixture. The antibody agglutinated all red cells except 6 examples of the 'Bombay' phenotype included in the panel. The antibody was neutralized by H secretor saliva, and its activity was abolished by DTT reagent, indicating its possible IgM nature. CONCLUSION: These findings suggest that this anomalous antibody has anti-H specificity. PMID- 9228714 TI - Gel test application for IgG subclass detection in auto-immune haemolytic anaemia. AB - BACKGROUND AND OBJECTIVES: Commercially available gel test microtubes are not available with antisera for the determination of IgG subclasses. The aim of this study was to adapt the gel technique for this purpose and apply it to the investigation of patients with autoimmune haemolytic anaemia (AIHA). MATERIALS AND METHODS: We studied 66 red cell samples from 49 patients with AIHA of the warm-active IgG type. Standard serologic and haematologic methods were used. We adapted the DiaMed gel test by using IgG-subclass antisera. RESULTS: We found the adapted test useful in determining the subclass of autoantibodies in eluates. We could identify the IgG subclass in all the AIHA patients, even those that were 'Coombs-negative', with less than 200 IgG molecules bound in vivo per red cell. Comparison of the gel test with the standard spin tube test and the microtiter plate test showed the superiority of the gel test, i.e., detection of IgG subclasses was much better than with the tube test and the results were more clearcut than those of the microplate test. The gel test is also the simplest and least time-consuming and permits a later reading of results. Application of the test in our 66 samples confirmed that IgG1 was the most frequent (96%). In 59% of the cases it was accompanied by IgG of other subclasses. Multiple subclasses were most common in the cases with stronger in vivo IgG red cell sensitization and severe haemolysis. Accompanying IgG3 was detected only in patients with obvious haemolysis. CONCLUSION: The gel test is more sensitive than other procedures for the determination of IgG subclass and has the advantages of simplicity, rapidity, low cost, and stability of the agglutinates. PMID- 9228715 TI - Platelet alloimmunization after transfusion. A prospective study in 117 heart surgery patients. AB - BACKGROUND AND OBJECTIVES: The frequency of platelet-specific antibodies after one series of blood transfusions has not been reported, and in multiply transfused patients is controversial. MATERIALS AND METHODS: We studied the frequency of alloimmunization against platelet antigens in 117 patients who received a single series of blood transfusions. They received mostly saline adenine-glucose+mannitol red blood cell components (poor in leukocytes and platelets) in connection with cardiac surgery. Platelet-specific antibodies were detected with the platelet ELISA and the monoclonal-antibody-specific immobilization of platelet antigen assay. HLA antibodies were detected by the standard lymphocyte cytotoxicity techniques. RESULTS: We found platelet-specific anti-HPA-5b (anti-Bra) in 2 cases (1.7%). One antibody was the result of de novo immunization. We detected lymphocytotoxic HLA antibodies in 21 patients (17.9%), of whom 18 (15.4%) had had no detectable antibodies before transfusion. There was a positive correlation between the transfused load of immunogenic materials and the frequency of alloimmunization against HLA antigens. In one third of the immunized patients, there was no history of previous immunization. CONCLUSION: There was a low incidence of platelet-specific antibodies after one series of blood transfusions in this group of patients. This is similar to the results of some previous studies in multiply transfused patients, but not with those of others who found a higher incidence. PMID- 9228716 TI - Platelet antibody detection in pediatric immune thrombocytopenic purpura: evaluation of three screening methods. AB - BACKGROUND AND OBJECTIVES: Immune thrombocytopenic purpura (ITP) is a common hematologic disorder, two forms of which occur in children. The detection of circulating platelet antibodies is helpful in diagnosis. MATERIALS AND METHODS: We evaluated three different immunological methods for detecting platelet antibodies in the serum of children with ITP. These were: a solid-phase red-cell adherence test (SPRCA), an enzyme immunoassay (EIA), and an immunofluorescence test (PSIF). RESULTS: The sensitivity of the methods in detecting IgG antibodies ranged from 28.1 (EIA) to 39.4% (SPRCA). We also looked for IgM antibodies by PSIF, thus raising the sensitivity of this test from 32.0 to 40.0%. A combination of two tests (SPRCA and EIA) allowed us to detect 61.8% positive samples. By doing all three tests, we obtained 71.3% positive samples. Finally, we reached 73.5% by adding PSIF for IgM. We found a higher frequency of circulating antibodies in both acute and chronic ITP at onset than in clinical remission. There were a few positive sera in chronic ITP, but not in the acute form in remission. CONCLUSION: The individual tests each have a relatively low sensitivity, but the combination of all three increases the diagnostic effectiveness. The finding of platelet antibodies during remission may predict evolution toward a systemic autoimmune state. PMID- 9228717 TI - Increased in vitro immunosuppressive action of anti-CMV and anti-HBs intravenous immunoglobulins due to higher amounts of interferon-gamma specific neutralizing antibodies. AB - BACKGROUND AND OBJECTIVES: We previously found that interferon-gamma (IFN-gamma) antibodies in intravenous immunoglobulins (IVIG) can block not only IFN-gamma production and tumor necrosis factor-alpha secretion, but also T-cell proliferation. Since the presence of IFN-gamma antibodies has been attributed to previous viral infection, we hypothesized that the viral status of the plasma donors used for IVIG pools might be a decisive factor in controlling the immunosuppressive capacity of IVIG. MATERIALS AND METHODS: We tested three different pooled, human IVIG preparations for the presence of IFN-gamma antibodies by ELISA. RESULTS: Comparison of the immunomodulatory activity of polyvalent IVIG with that of specific CMV and HBs IVIG showed that the latter-had higher levels of IFN-gamma antibodies and an increased capacity to block mixed lymphocyte reaction and cytokine production. CONCLUSION: We propose that these in vitro assays constitute a basis for the selection of plasma intended for manufacturing IVIG aimed at immunosuppression in the transplant setting. PMID- 9228718 TI - Transfusion support for a patient with severe common variable immunodeficiency with anti-IgA undergoing orthotopic liver transplantation. PMID- 9228719 TI - Perceptions of the risk of HIV infection through giving blood in Greece. PMID- 9228720 TI - Liquid pasteurization of immunoglobulins without stabilizer. PMID- 9228724 TI - Prospective therapeutic studies in nephrolithiasis. AB - Prospective randomized trials with respect to stone prophylaxis deal mainly with idiopathic calcium stone disease. There is a lot of evidence that alkali citrate, thiazides and allopurinol are effective in many patients. However, appropriate patient selection seems to be crucial. Alkali citrate and allopurinol have proven clinical efficacy in patients with hypocitraturia or hyperuricosuria, respectively. The correct method of patient selection for treatment with thiazides remains unclear. Despite the lack of prospective randomized trials, it is generally accepted that the first steps in the prophylaxis of idiopathic calcium stone disease are high fluid intake and a sensible diet. PMID- 9228723 TI - Current aspects of epidemiology and nutrition in urinary stone disease. AB - Current examples for the development of urinary stone disease are discussed by means of data from the literature and our own studies. Urinary stone disease has gained increasing significance due to changes in living conditions, i.e., industrialization and malnutrition. Changes in prevalence and incidence, the occurrence of stone types and stone location, and the manner of stone removal are explained. The importance of nutrition in the prevention of calcium oxalate stone disease is discussed in terms of fluid intake, calcium and oxalate metabolism, and dietary fat intake. The results of a study on a standardized mixed diet or an ovo-lactovegetarian diet show that well-balanced nutrition with consecutive high intake of fluids leads to a significant decrease in the risk for urinary stone formation (calculated as relative supersaturation with calcium oxalate by the computer program EQUIL). PMID- 9228722 TI - Urinary inhibitors of calcium oxalate crystallization and their potential role in stone formation. PMID- 9228725 TI - Risk formulas in calcium oxalate urolithiasis. AB - In order to reflect the risk of calcium stone formation, risk formulas have been described in the literature with the objective of being able to predict the further course of the stone disease. Some of these formulas are reviewed in this paper. Various results were obtained when different risk expressions were related to the severity of the stone disease. Although a reliable prediction of the future course of the disease most certainly cannot be made by analysis of the variables included in these expressions, several of the risk formulas differed significantly between patients with and without recurrent stone formation during a reasonable follow-up period. Some risk formulas might thus be helpful, at least to some extent, in selecting those patients in whom continuous stone formation can be anticipated and in whom active therapeutic measures should be beneficial and worthwhile. With an increased understanding of the mechanisms of calcium oxalate stone formation and our possibilities of measuring the relevant risk factors, it is likely that improved risk formulas with an increased predictive power can be developed. Until this becomes a reality, in most cases we have to combine important information on the history and clinical observations of the disease with a risk formula that offers a high degree of discrimination with respect to the risk of further stone formation. PMID- 9228726 TI - Genes in idiopathic calcium oxalate stone disease. AB - An examination of the urinary excretions of 101 normal subjects indicated that the major genetic influence on calcium excretion is a codominant pair of alleles giving rise to three phenotypes, low, intermediate and high (hypercalciuric) excretors. This inference was based on variance, Hardy-Weinberg and segregation analyses. Similar independent gene pairs also appear to influence oxalate and citrate excretion, A 3-locus Hardy-Weinberg table using estimates of gene frequencies derived from the study of normals suggests that only 3 or 4 leading genes are involved in oxalate stone disease. Strong candidate genes identified from molecular and physiological studies cannot be proposed at present, but it is assumed that they influence the transport of these ions in either the intestine, kidney or both organs. The identification of the genes involved should be facilitated by the reduction of dietary influences on urinary excretions through the use of formula diets. PMID- 9228727 TI - Treatment update on pediatric urolithiasis. AB - At the doorstep of the twenty-first century the role of extracorporeal shock-wave lithotripsy (ESWL) as the treatment of choice for more than 80% of all stones in children is established. ESWL is safe and effective, with very few differences in success rates being observed among different lithotriptors. The present problem with ESWL appears to be the residual stone fragment, which has a proven clinical significance. A thorough metabolic evaluation and metaphylaxis is indicated in all children, and this will enable physicians to deal with the residual fragments in a more cause-specific manner and prevent regrowth. Another subject that needs prospective randomized studies to be unveiled is the assumption that a specific or universal metaphylaxis, possibly with alkaline citrates, will enhance stone clearance, lower the incidence of residual stone fragments, and optimize the ESWL results. Finally, both percutaneous nephrolithotomy and ureteroscopic stone removal have been established in children as safe and effective treatment options. This gives the clinician the opportunity to choose from a wide range of treatment alternatives, including open surgery, and only this approach will ensure 100% stone removal in individual patients along with the prevention of recurrence and, thus, the elimination of long-term morbidities in this vulnerable patient population. PMID- 9228729 TI - A tryptophan-2-monooxygenase based amperometric biosensor for L-tryptophan determination: use of a competitive inhibitor as a tool for selectivity increase. AB - A new flow-injection amperometric biosensor based on immobilized tryptophan-2 monooxygenase (TMO) has been developed for reagentless L-tryptophan determination. Concentrations of L-tryptophan between 0.1 and 50 mM could be measured with the linear part of the calibration curve between 0.1 and 2 mM. The response time was 30 s and the total analysis time was less than 3 min. The biosensor retained activity for greater than 4 months, when operated daily at 25 degrees C and stored at 8 degrees C. The biosensor was characterized by a relatively high sensitivity to phenylalanine (54% that of L-tryptophan), a modest response to L-methionine (less than 6%) and virtually no response to other amino acids. However, the biosensor selectivity to L-tryptophan could be dramatically increased when indoleacetamide (IA), a competitive inhibitor of TMO, was introduced. In the presence of 10 microM IA, the biosensor response to L phenylalanine decreased to 7-4% of the unaffected rate for L-tryptophan. In the absence of L-tryptophan and IA the biosensor could be used for L-phenylalanine determination in the concentration range from 1 to 50 mM. The biosensor was successfully used for L-tryptophan determination in nutritional broth. PMID- 9228728 TI - Multiple primary tumors: 17 cases of renal-cell carcinoma associated with primary tumors involving different steroid-hormone target tissues. AB - The aim of this study was to analyze the characteristics of 17 women with renal cell carcinoma (RCC) associated with other primary neoplasms occurring in steroid hormone target tissues. The reproductive history of these patients and the use of exogenous hormones were taken into consideration. In all, 10 RCCs were associated with breast carcinoma; 4, with endometrial carcinoma; and 3, with ovarian carcinoma. The presentation of a second primary tumor was described as synchronous or metachronous by evaluation of the interval between the discovery of the two neoplasms. Hormone and surgical treatment as well as pathologic findings for each primary tumor were also reported. In these 17 RCCs the overall rate of disease-specific survival recorded after a mean follow-up period of 32.7 months (range 9-66 months) was 58.8%; 7 patients died of metastatic disease after surviving for a mean of 14.7 months. In terms of the pathologic stage of RCC, a significant difference in mean survival was found between pN0 (mean survival 22.1 +/- 3.4 months) and pN1 RCCs (mean survival 13.7 +/- 3.5 months). A total of 13 (76.4%) women were postmenopausal at the time of diagnosis of the first primary tumor; the mean age of these women at menopause was 51.7 +/- 1.2 years. No prior use of oral contraceptives was reported by 12 (70.5%) of the 17 patients. Plasma 17-beta-estradiol and estrone levels were determined in only 7 patients at the diagnosis of each of the primary tumors. High plasma estrogen levels were found in 4 women with RCC and breast carcinoma and in 1 patient with RCC and endometrial carcinoma; in the remaining 2 patients low-normal values were found. A relationship appears to exist between certain reproductive and hormone-related factors and the risk of developing these tumor associations. Data emerging from the present study do not provide strong support for the hypothesis of hormone dependency of RCC; however, a role for estrogens in cases in which RCC is associated with other primary tumors involving steroid-hormone target tissues can be hypothesized. PMID- 9228730 TI - Odor, drug and toxin analysis with neuronal networks in vitro: extracellular array recording of network responses. AB - Neurons, by virtue of intrinsic electrophysiological mechanisms, represent transducers that report the dynamics of cell death, receptor-ligand interactions, alterations in metabolism, and generic membrane perforation processes. In cell culture, mammalian neurons form fault-tolerant, spontaneously active systems with great sensitivity to their chemical environment and generate response profiles that are often concentration- and substance-specific. Changes in action potential patterns are usually detected before morphological changes and cell damage occur, which provides sensitivity and reversibility. Such biological systems can be used to screen rapidly for novel pharmacological substances, toxic agents, and for the detection of certain odorants. Existing simple culture preparations can already be employed effectively for the detection of chemical compounds. So far, three strategies have been investigated in pilot experiments: (1) Substance-dependent major changes in spontaneous native activity patterns. All synaptically active agents (e.g. glutamate, strychnine, N-methyl D-aspartic acid) as well as metabolic poisons generate such changes. (2) Substance-dependent changes in network oscillations via disinhibition. The regularized, oscillatory activity is altered by synaptically and metabolically active substances, ion channel blockers, and toxins. (3) Detection of paroxysmal responses indicating major, pathological membrane currents in large subpopulation of cells. We have explored these three strategies via 64 channel array recordings using spontaneously active murine spinal cord cultures. The glycine receptor blocker strychnine reliably generated increased multichannel bursting at 5-20 nM and regular, coordinated bursting above 5 microM. During biculline-induced network oscillations many compounds alter oscillation frequencies or terminate activity in a substance specific manner. Finally, the gp120 protein of the AIDS virus (at 1 microgram/ml) produces massive, unique paroxysmal discharges that may last as long as 2 min. These results indicate that cultured neuronal networks are practical systems that can be used for the detection and identification of a great variety of chemical substances. The concept of dynamic fingerprinting to identify specific compounds is discussed. PMID- 9228731 TI - Studies on dynamic behavior of the biosensor based on immobilized glucoamylase glucose oxidase membrane. AB - Frequency response of the glucose/maltose biosensors based on immobilized glucoamylase and glucose oxidase membranes of different thickness were investigated with the sinusoidal signal generator of substrate concentration. The results were modeled by the serial combination of the dead time and the first order response blocks, and time constants and dead times were estimated. The estimated value of time constant ranged from 4.5 to 13.2 s and that of the dead time from 3.2 to 7.7 s with the tendency that both time constant and dead time increase as membrane thickness increases and number of enzymatic reaction steps increases. PMID- 9228732 TI - Adsorption of immunoglobulin G on carbon paste electrodes as a basis for the development of immunoelectrochemical devices. AB - The attachment of Immunoglobulin G (IgG) to a carbon paste electrode is investigated in this work. Studies of the immobilization of the immunoglobulin on this electrode were carried out. Alkaline phosphatase (AP) has been used as an enzyme label, linked to the antibody, for the determination of the adsorbed immunoglobulin. Various substrates for this enzyme have been tested and alpha naphthyl phosphate was found to be the best because of the lower oxidation potential of the enzymatic product, alpha-naphthol, and greater sensitivity under the same experimental conditions. An electrodic pretreatment based on an anodic oxidation of the electrode surface in a phosphate media pH 9 was carried out prior to the adsorption step. This activation is also suitable for the removal of the IgG layer and the regeneration of the electrodic surface after each determination, with the intention of using the same electrode in the subsequent assays. A good reproducibility of the signal was achieved in this way (Relative Standard Deviation, R.S.D. = 3.7%). Using cyclic voltammetry, IgG labelled with AP has been quantified in a range from 2 x 10(-12) to 7 x 10(-11) M and a detection limit of 2.3 x 10(-12) M (signal-to-noise ratio = 3) was found. Finally, human IgG was quantified under non-optimized conditions on the electrodic surface through the reaction with AP labelled IgG using two different immunological designs. PMID- 9228733 TI - Flow-through amperometric immunosensor: fast 'sandwich' scheme immunoassay. AB - A flow-through immunosensor based on a high-surface-area carbon immunoelectrode has been developed. Dispersed carbon material serves as a carrier for immobilized antibodies and at the same time as an electrode material. The 'sandwich' scheme of immunoassay has been used. Iodine formed as a result of the enzymatic oxidation of iodide by a peroxidase label has been detected amperometrically. The immunosensor consists of a disposable sensing element (immunocolumn) containing dispersed carbon material with immobilized antibodies which also acts as a working electrode. A current collector connects the working electrode to the measuring device. The electrochemical detection time of the peroxidase-labeled immuno-complex does not exceed several minutes. The overall time of analysis including flowing of analyte, flowing of antigen, washing and detection stages is as low as 22 min. This technique allows fast determination of rabbit IgG (used as a model analyte) with a low detection limit in the picomolar range and also the determination of human IgM in blood plasma with a low detection limit in the nanomolar range. PMID- 9228734 TI - Electrochemical evaluation of chemical selectivity of glutamate receptor ion channel proteins with a multi-channel sensor. AB - A new method for evaluating chemical selectivity of agonists towards receptor ion channel proteins is proposed by using glutamate receptor (GluR) ion channel proteins and their agonists N-methyl-D-aspartic acid (NMDA), L-glutamate, and (2S, 3R, 4S) isomer of 2-(carboxycyclopropyl)glycine (L-CCG-IV). Integrated multi channel currents, corresponding to the sum of total amount of ions passed through the multiple open channels, were used as a measure of agonists' selectivity to recognize ion channel proteins and induce channel currents. GluRs isolated from rat synaptic plasma membranes were incorporated into planar bilayer lipid membranes (BLMs) formed by the folding method. The empirical factors that affect the selectivity were demonstrated: (i) the number of GluRs incorporated into BLMs varied from one membrane to another; (ii) each BLM contained different subtypes of GluRs (NMDA and/or non-NMDA subtypes); and (iii) the magnitude of multi channel responses induced by L-glutamate at negative applied potentials was larger than at positive potentials, while those by NMDA and L-CCG-IV were linearly related to applied potentials. The chemical selectivity among NMDA, L glutamate and L-CCG-IV for NMDA subtype of GluRs was determined with each single BLM in which only NMDA subtype of GluRs was designed to be active by inhibiting the non-NMDA subtypes using a specific antagonist DNQX. The order of selectivity among the relevant agonists for the NMDA receptor subtype was found to be L-CCG IV > L-glutamate > NMDA, which is consistent with the order of binding affinity of these agonists towards the same NMDA subtypes. The potential use of this approach for evaluating chemical selectivity towards non-NMDA receptor subtypes of GluRs and other receptor ion channel proteins is discussed. PMID- 9228735 TI - Chemoenzymatic synthesis of a trimeric ganglioside GM3 analogue. AB - A trimeric beta-lactosyl cluster based on 2-nitro-2-(hydroxymethyl)propane-1,3 diol was prepared using the trichloroacetimidate method. Kinetic studies showed that this cluster was an effective acceptor for rat-liver alpha-(2-->3) sialyltransferase. Its KM was comparable to those for monomeric lactose and N acetyllactosamine acceptors, and its Vmax was 1% of that measured for the LacNAc acceptor. Preparative-scale sialylation using this enzyme afforded a trimeric cluster of the ganglioside GM3 oligosaccharide in good yield. The NMR spectra of the trimeric GM3 analogue suggest that the oligosaccharide conformation is not significantly perturbed by this level of clustering. PMID- 9228736 TI - Thermodynamics of the hydrolysis and cyclization reactions of alpha-, beta-, and gamma-cyclodextrin. AB - A thermodynamic investigation of the hydrolysis and cyclization reactions of cyclomaltohexa-, hepta-, and octa-ose (alpha-, beta-, and gamma-cyclodextrins) has been performed using microcalorimetry and high-performance liquid chromatography. The calorimetric measurements lead to standard molar enthalpy changes delta rHm0 (T = 298.15 K, KH2PO4 buffer (m = 0.10 mol kg-1), pH = 4.58 to 5.15) for the following reactions: alpha-cyclodextrin(aq) + 6H2O(l) = 6 D glucose(aq), beta-cyclodextrin(aq) + 7H2O(l) = 7 D-glucose(aq), gamma cyclodextrin(aq) + 8H2O(l) = 8 D-glucose(aq). Equilibrium constants were determined for the following generalized cyclization reactions (T = 329.6 K, 0.005 mol kg-1 K2HPO4 buffer adjusted to pH = 5.55 with H3PO4) catalyzed by cyclomaltodextrin glucanotransferase: Gu(aq) = alpha-cyclodextrin(aq) + G(u 6)(aq), Gv(aq) = beta-cyclodextrin(aq) + G(v-7)(aq), Gw(aq) = gamma cyclodextrin(aq) + G(w-8)(aq). Here, G1 is D-glucose and the Gn's (n is a positive integer) are linear maltodextrins; u, v, and w are, respectively, integers > or = 7, > or = 8, and > or = 9. Values of the equilibrium constants, standard molar Gibbs energy change delta rGm0, standard molar enthalpy change delta rHm0, standard molar entropy change delta rSm0, and standard molar heat capacity change delta rCp,m0 are tabulated for the above reactions at T = 298.15 K. The values of delta rGm0 and delta rSm0 for the first three above-mentioned reactions rely upon an estimated value of delta rSm0 for the hydrolysis reaction of maltose to D-glucose. The thermodynamics of the disproportionation reaction Gm(aq) + Gn(aq) = Gm-1(aq) + Gn+1(aq) is also discussed. Values of the quantities delta rHm0/N, delta rGm0/N, delta rSm0/N, and delta rCp,m0/N for the three above mentioned hydrolysis reactions where N is the number of (1-->4)-alpha-D glucosidic bonds broken in each of these reactions, have been calculated and compared with thermodynamic quantities for the similar hydrolysis reaction of a linear oligosaccharide. PMID- 9228737 TI - Structure of the 13C-enriched O-deacetylated glucuronoxylomannan of Cryptococcus neoformans serotype A determined by NMR spectroscopy. AB - The complete assignment of 1H and 13C chemical shifts for 99% uniformly 13C labeled O-deacetylated glucuronoxylomannan (GXM) of Cryptococcus neoformans serotype A isolate 9759-Mu-1 was accomplished by the analysis of HCCH-TOCSY and HCCH-COSY spectra. The sequence of the glycosyl residues was determined by a GHMBC experiment using 20% uniformly 13C-labeled GXM; GXM was prepared by a novel procedure that insured the virtual exclusion of adjacent 13C-labeled carbon atoms. For each residue in the GXM of 9759-Mu-1 we determined its linkage position, its anomeric configuration, and its position in the repeating sequence as follows: [sequence: see text] PMID- 9228738 TI - Structural characterisation of the exocellular polysaccharide produced by Streptococcus thermophilus OR 901. AB - The exocellular polysaccharide of Streptococcus thermophilus OR 901, isolated from partially deproteinised whey, is a heteropolymer of D-galactopyranose and L rhamnopyranose residues in the molar ratio 5:2. The structure was established by methylation analysis and 1D and 2D NMR spectroscopy of the native polysaccharide, in combination with characterisation of oligosaccharide fragments, obtained by partial acid hydrolysis, using methylation analysis and 1D 1H NMR spectroscopy. The polysaccharide has a branched heptasaccharide repeating unit with the following structure: [sequence: see text] PMID- 9228739 TI - Structure of the O-specific polysaccharide of Salmonella enterica ssp. arizonae O50 (Arizona 9a,9b). AB - On the basis of sugar and methylation analysis, selective removal of 3,6-dideoxy L-xylohexose (colitose, Col), 1H and 13C NMR spectroscopy, including 1D NOE, 2D COSY, and 2D H-detected 1H, 13C heteronuclear multiple-quantum coherence (HMQC), the following structure of the repeating unit of the O-specific polysaccharide of Salmonella enterica ssp. arizonae O50 (Arizona 9a,9b) was established: [sequence: see text] The O-antigen studied includes a trisaccharide fragment alpha-Co1p-(1- >2)-beta-D-Galp-(1-->3)-beta-D-GlcpNAc, which is a colitose ('3-deoxy-L-fucose') analogue of the Lewis (precursor) blood group antigen. PMID- 9228740 TI - Development of an immunoassay for larch arabinogalactan and its use in the detection of larch arabinogalactan in rat blood. AB - We describe a sensitive and convenient immunoassay for larch arabinogalactan and demonstrate its specificity for larch arabinogalactan. Anti-larch arabinogalactan antiserum is about 10(4) and 10(6) times more selective for detecting larch arabinogalactan than antiserum binds to branch terminal disaccharides consisting of the terminal beta-D-galactosyl residue and the penultimate branch (1-->6)-beta D-galactosyl residue. It does not bind L-arabinose. The sensitivity of the assay for larch arabinogalactan is less than 0.1 microgram/mL. The application of the assay for measuring arabinogalactan pharmacokinetics in rat blood is illustrated. PMID- 9228741 TI - Carbon repression in Aspergilli. AB - Many microorganisms prefer easily metabolizable carbon sources over alternative, less readily metabolized carbon sources. One of the mechanisms to achieve this is repression of the synthesis of enzymes related to catabolism of the alternative carbon sources, i.e. carbon repression. It is now clear that in Aspergillus nidulans and Aspergillus niger the repressor protein CREA plays a major role in carbon repression. CREA inhibits transcription of many target genes by binding to specific sequences in the promoter of these genes. Unfortunately there is little information on other components of the signalling pathway that triggers repression by CREA. In this review we summarize the current understanding of carbon repression in Aspergilli. PMID- 9228742 TI - Function and modulation of bacterial porins: insights from electrophysiology. AB - Electrophysiological techniques provide a wealth of information regarding the molecular mechanisms that underlie the function and modulation of ion channels. They have revealed that bacterial porins do not behave as static, permanently open pores but display a much more complex and dynamic behavior than anticipated from non-electrophysiological studies. The channels switch between short-lived open and closed conformations (gating activity), and can also remain in an inactivated, non-ion conducting state for prolonged periods of time. Thus the role of porins is not limited to that of a molecular filter, but is extended to the control of outer membrane permeability through the regulation of their activity. Electrophysiological studies have indeed demonstrated that both gating and inactivation are modulated by a variety of physical and chemical parameters and are highly cooperative phenomena, often involving numerous channels working in concert. Cooperativity acts as an amplification mechanism that grants a large population of porins, such as found in the outer membrane, with sensitivity to modulation by external or internal factors. By conferring permeability properties to the outer membrane, porins play a crucial role in the bacterium's antibiotic susceptibility and survival in various environmental conditions. The detailed information that electrophysiology only can provide on porin function and modulation promises to yield a more accurate description of how porin properties can be used by cells to adapt to a changing environment, and to offer mechanisms that might optimize the drug sensitivity of the microorganism. PMID- 9228743 TI - Acyl phosphatidylglycerol: a major phospholipid of Corynebacterium amycolatum. AB - The type strain and several clinical isolates of Corynebacterium amycolatum were examined for lipid composition as a chemotaxonomic character for routine identification. The phospholipid profile was composed of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylinositol and phosphatidylinositol mannosides, together with various unidentified compounds. One of them, accounting for 20-29% of total phospholipids, was purified and characterized as acyl phosphatidylglycerol by chromatographic and spectrometric techniques. The acyl substituents on the phosphatidyl moiety were characterized as tetradecanoyl, pentadecanoyl, hexadecenoyl, hexadecanoyl, heptadecenoyl, heptadecanoyl, octadecenoyl (the major one), and octadecanoyl. The acyl group on the polar head (glycerol) was only octadecenoyl. Phospholipid analysis by thin-layer chromatography of a collection of Corynebacterium strains proved that this compound is widely distributed, although it only represents a minor (2-9%) component among mycolic acid-containing species. Acyl phosphatidylglycerol can be considered as a useful chemical marker for the identification of C. amycolatum in addition to the absence of mycolic acids. PMID- 9228744 TI - GAP1 activity is dependent on cAMP in Saccharomyces cerevisiae. AB - General amino acid permease (GAP1) activity was evaluated in adenylate cyclase deficient Saccharomyces cerevisiae to determine the effect of cAMP levels on GAP1 activity. Lowering cAMP concentrations in the culture media led to a decrease in the initial rates of L-citrulline uptake. Kinetics of the amino acid transport system showed a partial loss of transport capacity, with no apparent modifications in permease affinity. PMID- 9228745 TI - Electroporation of intact cells of Streptomyces parvulus and Streptomyces vinaceus. AB - Various assays of classical PEG-assisted transformation as well as electrotransformation of Streptomyces parvulus IMET41380 and Streptomyces vinaceus NCIB8852 are described. Contrary to the so far reported assays of electrotransforming Streptomyces strains, electroporation in S. parvulus and S. vinaceus was carried out on intact cells, without any lysozyme treatment. In these two strains, the classical PEG-assisted transformation of protoplasts does not work efficiently (10(3) to 10(4) transformants per micrograms of pIJ702 DNA) and electrotransformation gives 10 to 100 times higher yields (10(5) transformants per micrograms of pIJ702 DNA). The electroporation method described here is not applicable to other Streptomyces strains (S. lividans or S. coelicolor). PMID- 9228746 TI - Distribution of the synergistic haemolysin genes hld and slush with respect to agr in human staphylococci. AB - Several staphylococcal species express a synergistic activity which potentiates hemolysis by beta-hemolysin. In Staphylococcus aureus, this activity is mediated by the delta-hemolysin, coded by the hld gene within the agr locus. In S. lugdunensis, the equivalent activity results from the production of 3 small peptides coded by an operon named slush, distinct from hld and located outside the agr region. We examined 15 clinically relevant staphylococcal species for the presence of hld, slush and agr by specific hybridisation. All species contained a recognizable agr-related locus. Three species never produced synergistic haemolysis and contained neither hld nor slush. Of the 12 producer strains, 5 contained hld, apparently within the agr region, 4 contained slush and one (S. caprae) contained both. Two other producer species (S. hominis and S. simulans) hybridised to neither probe. PMID- 9228747 TI - 16S rRNA gene sequence analyses and inter- and intrageneric relationships of Xanthomonas species and Stenotrophomonas maltophilia. AB - The nearly complete, PCR-amplified, 16S rRNA gene sequences have been determined from the representative type strains of eight xanthomonad phena, including six validly described species of the genus Xanthomonas and Stenotrophomonas maltophilia. Pairwise sequence comparisons and phylogenetic analysis demonstrated that the xanthomonads comprise a monophyletic lineage-within the gamma-subclass of the Proteobacteria. Although the genus Xanthomonas was observed to comprise a cluster of very closely related species, the observed species-specific primary sequence differences were confirmed through sequencing additional strains belonging to the respective species. PMID- 9228749 TI - Effect of the glutamine synthetase inhibitor, methionine sulfoximine, on the growth and differentiation of Dictyostelium discoideum. AB - To examine further the role of the enzyme glutamine synthetase in Dictyostelium discoideum we report here the effects of a specific glutamine synthetase inhibitor, methionine sulfoximine, on the growth and differentiation of this organism. Vegetative AX3 cells grown in the presence of methionine sulfoximine did not complete culmination in the normal time but were blocked at the finger stage. In these cells glutamine synthetase activity was almost completely abolished. However, methionine sulfoximine did not affect the level of glutamine synthetase mRNA, suggesting that there is no link between glutamine synthetase activity and mRNA transcription. Eventually glutamine synthetase activity reappeared and at the time culmination occurred. These results suggest that glutamine synthetase plays an important role in the assimilation of ammonia during the later stages of development in D. discoideum and that this assimilation is necessary for the completion of culmination. PMID- 9228748 TI - Restriction fragment length polymorphisms of 16S rDNA and of whole rRNA genes (ribotyping) of Streptococcus iniae strains from the United States and Israel. AB - Streptococcus iniae (junior synonym S. shiloi) isolated from tilapia and trout in Israel and in the United States were subtyped by restriction length polymorphism (RFLP) based on PCR amplified 16S rDNA and by ribotyping. 16S rDNA RFLP discriminated between S. iniae and other fish pathogens but not between S. iniae strains. HindIII and EcoRI ribotypes of S. iniae discriminated American from Israeli strains rejecting the possibility of an epidemiological link between S. iniae infections in the two countries. Israeli strains isolated from tilapia and trout could not be completely differentiated. The S. iniae ATCC 29178T (T = Type strain) strain, isolated from a freshwater dolphin belonged to a ribotype different from those of all the fish isolates. PMID- 9228750 TI - Purification and partial characterization of an antigen specific to Lactobacillus brevis strains with beer spoilage activity. AB - Certain Lactobacillus brevis strains are resistant to hop-derived compounds such as isohumulone and are able to grow in beer. In this study, we raised an antiserum against our beer spoilage laboratory strain L. brevis 578 which reacted with 23 of 24 beer spoilers and two of 13 non-spoilers in precipitation reactions using 0.5 M NaOH cell extracts. This specific antigen to the beer spoilage L. brevis strains (SABSL) was demonstrated to be located beneath the S-layer proteins by agglutination reactions using S-layer protein-stripped cells obtained by treatment with 0.1 M NaOH. SABSL was purified using an affinity column coupled with an antibody against SABSL. The purified antigen was hydrolyzed with 2 M HCl and the hydrolyzate was analyzed by thin-layer chromatography and enzymatic analysis. The results showed that SABSL contains glycerol, phosphate, glycerophosphate, D-galactose and D-glucose. D-Galactose and D-glucose accounted for 4.7% and 0.1% of the composition, respectively. Melibiose, but not mannose, inhibited the precipitation reaction. Intense precipitation reactions were obtained with fractions which did not bind to the ConA-column. These results indicate that the immunodominant component of the SABSL is galactose and the SABSL determinant is most probably a galactosylated glycerol teichoic acid. The antiserum raised against the beer spoilage strain L. brevis 578 could distinguish between Pediococcus beer spoilers and non-spoilers in precipitation reactions. PMID- 9228751 TI - Evidence for regulation of the NADH peroxidase gene (npr) from Enterococcus faecalis by OxyR. AB - We report that the purified Escherichia coli OxyR protein can bind specifically upstream of the gene encoding NADH peroxidase (npr) from Enterococcus faecalis 10C1, to a site located some 144 bp from the promoter. A 34 kDa protein has been identified in crude extracts of E. faecalis that cross-reacts with polyclonal antisera to purified OxyR from E. coli and a protein(s) present in these extracts retards npr DNA fragments in gel shift assays. Taken together with the results of sequence analyses, these observations suggest that enterococcal npr is regulated by OxyR. PMID- 9228752 TI - Double-stranded origin nicking and replication initiation are coupled in the replication of a rolling circle plasmid, pT181. AB - The Staphylococcus aureus rolling circle plasmid pT181 initiator RepC is modified by the addition of an oligodeoxynucleotide, giving rise to a new form, RepC*. RepC/RepC* heterodimer is an inhibitor of replication. However, in order to act effectively, the initiator/inhibitor protein must be stable. We show here that RepC is stable for at least 90 min, which enables it to function effectively as an inhibitor of replication. This finding also allowed us to carry out the two stages in pT181 replication sequentially: first, binding/nicking of the double strand origin (DSO) by the pT181-encoded RepC, followed by initiation/elongation by the host cell's DNA replication apparatus. The results demonstrate that these two stages in pT181 replication are functionally coupled and that interruptions in this continuous process generate relaxed pT181 DNA that cannot be used as a template for replication. PMID- 9228753 TI - Stress proteins and stress tolerance in an Antarctic, psychrophilic yeast, Candida psychrophila. AB - Conditions are described for the heat shock acquisition of thermotolerance, peroxide tolerance and synthesis of heat shock proteins (hsps) in the Antarctic, psychrophilic yeast Candida psychrophila. Cells grown at 15 degrees C and heat shocked at 25 degrees C (3 h) acquired tolerance to heat (35 degrees C) and hydrogen peroxide (100 mM). Novel heat shock inducible proteins at 80 and 110 kDa were observed as well as the presence of hsp 90, 70 and 60. The latter hsps were not significantly heat shock inducible. The absence of hsp 104 was intriguing and it was speculated that the 110 kDa protein may play a role in stress tolerance in psychrophilic yeasts, similar to that of hsp 104 in mesophilic species. PMID- 9228754 TI - Isolation and characterization of the 54-kDa and 22-kDa chitinase genes of Serratia marcescens KCTC2172. AB - A DNA fragment (pCHI5422) containing two genes encoding a 54-kD and a 22-kDa chitinase was isolated from a cosmid DNA library of Serratia marcescens KCTC2172. The complete nucleotide sequence of pCHI5422 consisting of 4581 bp was determined. The nucleotide sequence of the 22-kDa chitinase consists of 681 bp of open reading frame encoding 227 amino acids and is located 1422 bp downstream of the translation termination codon of the 54-kDa chitinase sequence. The 54-kDa chitinase gene consisted of 1497 bp in a single open reading frame encoding 499 amino acids. The genes encoding the 54-kDa and 22-kDa chitinase were separately subcloned in Escherichia coli and the individual chitinases were expressed and purified from the culture broth using chitin affinity chromatography. When chitohexaose was used as substrate, the major product of the enzymatic reaction of both the 54-kDa and 22-kDa chitinases was a (GlcNAc)2 dimer with a minor amount of monomer. The specific activity of the 54-kDa and 22-kDa chitinases were 300 microM (min)-1 mg-1 and 17 microM (min)-1 mg-1 on the natural swollen chitin, respectively. PMID- 9228755 TI - Construction and characterization of a recombinant ureolytic Streptococcus mutans and its use to demonstrate the relationship of urease activity to pH modulating capacity. AB - To begin to understand the contribution of oral microbial ureolysis to the inhibition of dental caries, we sought to construct a recombinant, ureolytic mutans streptococcus and correlate the ureolytic capacity of plaque bacteria with pH moderating ability. Streptococcus mutans GS-5 was transformed with a plasmid containing the urease genes from Streptococcus salivarius 57.I. The recombinant strain, S. mutans AC04, stably maintained the urease genes. High levels of urease activity were detected, with a maximum specific activity of 0.9 mumol of urea hydrolyzed/min/mg cell dry weight when the growth medium was supplemented with 50 microM exogenous NiCl2. Harboring the recombinant plasmid, or growth in NiCl2, did not markedly affect the glycolytic capacity of S. mutans. In vitro pH drop analysis of S. mutans AC04, metabolizing glucose and physiologically relevant concentrations of urea simultaneously, demonstrated that increasing the urease activity of plaque bacteria resulted in a corresponding reduction in the depth and the duration of the glycolytic pH fall. The results demonstrate the feasibility of engineering urease producing S. mutans and suggest that enhancing the ureolytic capacity of dental plaque, particularly cariogenic plaque, may help to offset the progression of the caries process. PMID- 9228756 TI - The role of flagella and motility in the adherence and invasion to fish cell lines by Aeromonas hydrophila serogroup O:34 strains. AB - We compared the ability of Aeromonas hydrophila wild-type strains of serogroup O:34, non-motile Tn5 aflagellar mutants and the same mutants harboring a recombinant cosmid DNA from a library of A. hydrophila AH-3 (O:34, wild-type) that allows these mutants to make flagella and to be motile, to adhere and invade two fish cell lines. We found that motility is essential in these strains for adhesion, and also that possession of flagella is essential for the ability to invade the fish cell lines. We cannot rule out that flagella may be an adhesin, or that motility may also be involved in A. hydrophila serogroup O:34 bacterial invasion of both fish cell lines. PMID- 9228757 TI - Identification and characterization of thymidylate synthase from Neisseria gonorrhoeae. AB - Thymidylate synthase converts deoxyuridylate to deoxythymidylate. The thyA gene has been cloned and sequenced from Neisseria gonorrhoeae MS11. This gene has an open reading frame of 795-801 bp and encodes a product which shares 71% identity with its Escherichia coli homolog. Unlike its E. coli counterpart, gonococcal thyA has a large, upstream transcribed region (300+ bp) that lacks a translatable reading frame. Gonococcal thyA is capable of complementing an E. coli thyA null mutant and shares similar levels of sensitivity with trimethoprim. PMID- 9228758 TI - Identification of Pediococcus acidilactici and Pediococcus pentosaceus based on 16S rRNA and ldhD gene-targeted multiplex PCR analysis. PMID- 9228759 TI - Molecular characterization of the replicon of the Pediococcus pentosaceus 43200 pediocin A plasmid pMD136. AB - The pediocin A-encoding plasmid of Pediococcus pentosaceus 43200, pMD136, was characterized by restriction enzyme analysis. Analysis of its replicon was facilitated by the construction of a probe vector consisting of the Escherichia coli plasmid pSP72 and the cat gene from Staphylococcus aureus plasmid pC194. The replication region of pMD136 was localized on a 1.6-kb EcoRI/BglII fragment. Sequencing analysis revealed a non-coding region, repA, spanning the first 440 bp, followed by an open reading frame, repB, encoding a putative protein of 390 amino acids. The non-coding region contained two sets of 6-bp and two sets of 22 bp direct repeats and two sets of inverted repeats upstream of the open reading frame. Strong homology of the isolated replicon was found to theta-type replicons of Lactococcus lactis plasmids. Segregational stability assay suggested at least two regions as potentially involved in the stabilization of pMD136. The plasmid's strong homology to other theta-type replicons and its relatively high stability suggest that pMD136 belongs to the widespread family of theta-replication plasmids. PMID- 9228760 TI - Purification and characterization of L-allo-threonine aldolase from Aeromonas jandaei DK-39. AB - L-allo-Threonine aldolase (L-allo-threonine acetaldehyde-lyase), which exhibited specificity for L-allo-threonine but not for L-threonine, was purified from a cell-free extract of Aeromonas jandaei DK-39. The purified enzyme catalyzed the aldol cleavage reaction of L-allo-threonine (K(m) = 1.45 mM, Vmax = 45.2 mumol min-1 mg-1). The activity of the enzyme was inhibited by carbonyl reagents, which suggests that pyridoxal-5'-phosphate participates in the enzymatic reaction. The enzyme does not act on either L-serine or L-threonine, and thus it can be distinguished from serine hydroxy-methyltransferase (L-serine:tetrahydrofolate 5,10-hydroxy-methyltransferase, EC 2.1.2.1) or L-threonine aldolase (EC 4.1.2.5). PMID- 9228761 TI - Borrelia burgdorferi uridine kinase: an enzyme of the pyrimidine salvage pathway for endogenous use of nucleotides. AB - The 621 bp udk gene encoding Borrelia burgdorferi potential uridine kinase, involved in the pyrimidine salvage pathway, was cloned and sequenced. The B burgdorferi protein has a molecular mass of 24 kDa in sodium dodecyl sulfate polyacrylamide gel. The N-terminal sequence of the protein, Ala-Lys-Ile-Ile, is identical to that predicted but lacks N-terminal methionine. udk is located at around 15 kb from the left telomere and forms an operon with an upstream ORF. A likely hypothesis for the role of the pyrimidine salvage pathway is the sole use of endogenous nucleotides for Borrelia. PMID- 9228762 TI - Molecular analysis of cyp51 from fluconazole-resistant Candida albicans strains. AB - The target enzyme for fluconazole is sterol 14 alpha-demethylase, a cytochrome P450 encoded by cyp51. One mechanism of fluconazole resistance likely to occur in Candida albicans is through an altered target site. To test this hypothesis DNA sequencing of the cyp51 coding sequence from 19 fluconazole-resistant and 19 fluconazole-sensitive C. albicans was undertaken. A number of point mutations were identified in the resistant isolates which were not present in the sensitive ones: F105L (five), E266D (five), K287R (one), G448G (one), G450E (one), G464S (three) and V488I (one). These alterations are discussed in the light of a molecular model of the enzyme regarding potential roles in resistance. It was also demonstrated that sequence-specific primers can be employed to identify polymorphisms which may be associated with resistance; diagnostic tests for resistant strains will prove of value in combating this serious clinical problem. PMID- 9228763 TI - Secreted enzymes of Aeromonas. AB - A hallmark characteristic of species of Aeromonas is their ability to secrete a wide variety of enzymes associated with pathogenicity and environmental adaptability. Among the most intensively studied are beta-lactamases, lipases, hemolytic enterotoxins, proteases, chitinases, nucleases and amylases. Multiple copies of genes encoding each type of enzyme provide additional biological diversity. Except for the chitinases, these multiple copies show little evolutionary relatedness at the DNA level and only limited similarity at the protein level. Indeed a number of the genes, such as nuclease H of A. hydrophila, have no similarity to known prokaryotic or eukaryotic sequences. The challenge is to determine how these genes evolved, where they originated and why Aeromonas possesses them in such abundance and variety. PMID- 9228764 TI - A possible role of trehalose in osmotolerance and ethanol tolerance in Saccharomyces cerevisiae. AB - The effect of salt stress on ethanol endurance of yeast cells was studied. Cells grown under increased NaCl concentrations were more ethanol tolerant than controls. The increase in trehalose content under hyper-saline conditions has been suggested to allow cells to withstand higher ethanolic conditions. There seems to be an overlap between osmotolerance and ethanol endurance in Saccharomyces cerevisiae. PMID- 9228765 TI - Effect of hydrostatic pressure of a mutant of Saccharomyces cerevisiae deleted in the trehalose-6-phosphate synthase gene. AB - Mutants Saccharomyces cerevisiae deleted on the trehalose-6-phosphate synthase gene (tps1) and their parental wild-type cells were submitted to hydrostatic pressure in the range of 0-200 MPa. Experimental evidence showed that viability for both strains decreased with increasing pressure and that tps1 mutants, unable to accumulate trehalose, were more sensitive to hydrostatic pressure than the wild-type cells. Additionally, both tps1 and wild-type cells in the stationary phase, when there is an accumulation of endogenous trehalose, were more resistant to pressure than proliferating cells. Under these conditions, mutant cells were also more sensitive to pressure treatment than the wild type. The present work also showed that mild pressure pretreatment did not induce hydrostatic pressure resistance (barotolerance) in yeast cells. PMID- 9228766 TI - Rapid purification of an active recombinant His-tagged 2,3-dihydroxybiphenyl 1,2 dioxygenase from Pseudomonas putida OU83. AB - 2,3-Dihydroxybiphenyl 1,2-dioxygenase (2,3-DBPD) is an extradiol-type dioxygenase that catalyzes the aromatic ring fission of 2,3-dihydroxybiphenyl, the third step in the biphenyl degradation pathway. The nucleotide sequence of the Pseudomonas putida OU83 gene bphC, which encodes 2,3-DBPD, was cloned into a plasmid pQE31. The His-tagged 2,3-DBPD produced by a recombinant Escherichia coli strain, SG13009(pREP4)(pAKC1), and purified with a Ni-nitrilotriacetic acid resin affinity column using the His-bind Qiagen system. The His-tagged 2,3-DBPD construction, carrying a single 6 x His tail on the N-terminal of the polypeptide, was active. SDS-PAGE analysis of the purified active 2,3-DBPD gave a single band of 34 kDa; this is in agreement with the size of the bphC coding region. The K(m) for 2,3-dihydroxybiphenyl was 14.5 +/- 2 microM. The enzyme activity was enhanced by ferrous ion but inhibited by ferric ion. The enzyme activity was inhibited by thiol-blocking reagents and heavy metals HgCl2, CuSO4, NiSO4, and CdCl2. The yield was much higher and the time required to purify recombinant 2,3-DBPD from clone pAKCl was faster than by the conventional chromatography procedures. PMID- 9228767 TI - Phylogeny of the genus Chlorobium based on 16S rDNA sequence. AB - A comparative analysis of 16S ribosomal DNA sequences from 18 strains belonging to different species of the genus Chlorobium has been made in order to investigate their phylogenetic relationships. All studied strains with brown pigmentation, belonging to species C. phaeobacteroides and C. phaeovibrioides, were clustered together and separated from green species. Most of the C. limicola strains formed a consistent group and were clustered next to C. tepidum. But, despite their identical morphology, C. vibrioforme strain 6030 (= DSM 260T) and C. vibrioforme strain 8327 (= DSM 263) were clearly differentiated on the basis of their 16S rDNA sequence. On the other hand, morphologically distinctive strains of C. chlorovibrioides, C. vibrioforme and C. limicola presented sequences with a high degree of similarity. Since morphological characters do not seem to be sufficient to adequately classify this group, these findings help to set up the basis for a revision of the taxonomy of the genus Chlorobium. PMID- 9228768 TI - T-track PCR fingerprinting for the rapid detection of genetic polymorphism. AB - The diversity of DNA sequences can be analyzed by comparing randomly amplified polymorphic DNA, or restriction fragment length polymorphism fragments of DNA. Such analyses are dependent on the selection of appropriate restriction enzyme(s) and/or primers. We have investigated a simpler approach to providing sensitive and specific genotyping. Cyclic extension of target sequences with dideoxythymidine generates PCR products with variable lengths. We analyzed these variable PCR products by scoring the number of variable bands and comparing the scores (numerical profiles) to establish similarities. We found that the polymorphic lengths of the PCR products were comparable among serologically defined strains. It suggests that this single PCR reaction followed by a one-step electrophoresis yields easily analyzable data that can be compared with data from other gels. PMID- 9228769 TI - Partially oxidized polycyclic aromatic hydrocarbons show an increased bioavailability and biodegradability. AB - Polycyclic aromatic hydrocarbons have a low water solubility and tend to adsorb on soil particles, which both result in slow bioremediation processes. Many microorganisms, known for their ability to degrade polycyclic aromatic hydrocarbons, only partially oxidize these compounds. White rot fungi, for instance, convert polycyclic aromatic hydrocarbons to more water soluble and bioavailable products. Polycyclic aromatic hydrocarbon metabolites were more readily mineralized by natural mixed bacterial cultures, like activated sludge and soil, than the parent polycyclic aromatic hydrocarbon compounds. These results suggest that sequential breakdown by white rot fungi followed by indigenous bacteria leads to an effective polycyclic aromatic hydrocarbon bioremediation process. PMID- 9228770 TI - Functional interactions between homologous conditional killer systems of plasmid and chromosomal origin. AB - parD and chpA are homologous conditional killer systems of plasmid and chromosomal origin, respectively, encoding a killer protein (Kid and ChpAK) and an antidote (Kis and ChpAI). Here it is shown that these systems can functionally interact. A multicopy chpA recombinant partially complements two mutations in the antidote of the parD system. These mutations affect either autoregulation or neutralization of the killer component. Following in vitro mutagenesis with hydroxylamine, chpA mutants that improve this complementation were isolated. Sequence analysis shows that these mutants are clustered in the 5' end of the chpAI gene and structure predictions suggest that they affect a putative loop in the secondary structure of the ChpAI antidote. It is proposed that this region is part of a protein-protein interface required for the functional interaction between the antidote and the killer components in the two homologous systems. PMID- 9228772 TI - Improved methods for in situ enzymatic amplification and detection of low copy number genes in bacteria. AB - We present alternative and improved protocols for in situ analysis of single copy genes in prokaryotes. Primed in situ amplification (PRINS) and cycle PRINS were used to detect, via the incorporation of a fluorescein labelled nucleotide, the presence of specific genes carried on both high and low copy number plasmids in individual cells of Escherichia coli and a marine bacterium, SW5. The optimised protocols described enabled a significant reduction in non-specific signals whilst maintaining high fluorescent activity via labelled nucleotide incorporation. In addition, nucleic acids were amplified linearly and were retained within the permeabilised microbial cells. These methods provide considerable advances in sensitivity, specificity and reliability compared to current protocols for bacterial in situ nucleic acid amplification. PMID- 9228771 TI - The general amino acid permease of Rhizobium leguminosarum strain 3841 is negatively regulated by the Ntr system. AB - Cosmid-borne and chromosomal lacZ fusions to aapJ. aapQ and aapM were used to examine the nitrogen regulation of the general amino acid permease (Aap) of Rhizobium leguminosarum strain 3841. Transcription of the first gene of the operon (aapJ), which encodes the periplasmic binding protein, was 2-4-fold higher than aapQ and aapM, which encode the integral membrane proteins, under various growth conditions. This may be due to the presence of a putative stem loop in the intergenic region between aapJ and aapQ. All aap fusions were derepressed 3-5 fold after growth on glutamate as a nitrogen source, which effectively causes nitrogen limitation. An ntrC mutant was derepressed for transcription of the aap operon and had high rates of amino acid transport when grown on ammonia as the nitrogen source. Thus NtrC negatively regulates the aap operon, contrary to its usual role in positive gene activation. These results confirm that the aap-operon is subject to complex regulation involving both transcriptional and post transcriptional factors. PMID- 9228773 TI - Ornithine cycle in Nostoc PCC 73102: presence of an in vitro functional argininosuccinate lyase. AB - The presence of argininosuccinate lyase (ASL), an enzyme catalyzing the final step of arginine biosynthesis, was demonstrated in the heterocystous cyanobacterium Nostoc PCC 73102 by measuring in vitro enzyme activity and by visualization of ASL in native protein gels. Activity staining of a native PAGE gel revealed one ASL-dependent band with a molecular mass of about 240 kDa. A colorimetric assay for ASL based on the measurement of urea produced from arginine in the presence of an excess of arginase was further used to analyze the cyanobacterial ASL. The in vitro ASL activity was highest in cells in exponential growth phase and decreased significantly during the stationary phase of growth, ranging from 4.4 to 0.8 nmol of product formed (mg protein)-1 min-1. Including arginine, citrulline or ornithine in the growth medium resulted in no significant change in the ASL activities, indicating that Nostoc PCC 73102 ASL is not regulated by metabolites of the ornithine cycle. PMID- 9228774 TI - Nuclear translocation of constitutive heat shock protein 70 during S phase in synchronous macroplasmodia of Physarum polycephalum. AB - The level of constitutive heat shock protein 70 (HSC70) in Physarum polycephalum was analyzed by means of Western blots during the synchronous cell cycle of macroplasmodia. Total amounts as well as nuclear and cytoplasmic contents were determined separately and evaluated densitometrically. A drastic increase of nuclear HSC70 was observed 10-40 min after the initiation of S phase (600% of the M phase value) and thereafter a slow decline toward the next M phase. Total HSC levels showed a slight (30%) increase during S phase whereas cytoplasmic HSC70 was about 30% lower during S phase compared to mitosis. PMID- 9228775 TI - The traditional enteropathogenic Escherichia coli (EPEC) serogroup O125 comprises serotypes which are mainly associated with the category of enteroaggregative E. coli. AB - Genotypic and phenotypic virulence markers of the different categories of diarrheagenic Escherichia coli were investigated in 76 strains of the enteropathogenic E. coli (EPEC) serogroup O125. The most frequent serotype found was O125ac:H21. None of the serotypes behaved as EPEC, i.e. carried the eaeA, bfpA, and EAF DNA sequences simultaneously and presented localized adherence to HeLa cells. All strains of O125ac:H6 were atypical EPEC since they carried eaeA only, and presented an indefinite pattern of adherence. All strains of O125ab:H9, O125ac:H9, O125?:H16, and O125ab:H21 and 79% of the O125ac:H21 strains were enteroaggregative E. coli, since they carried a specific DNA sequence and presented the typical aggregative adherence pattern. PMID- 9228776 TI - The adhesin related 30-kDa protein of Mycoplasma pneumoniae exhibits size and antigen variability. AB - Mycoplasma pneumoniae is a pathogenic bacterium colonizing epithelial cells of the human respirator tract. Using an erythrocyte binding assay we isolated a cytadsorption negative mutant designated M7 which has lost 12 of a total of 13 repetitive sequences of a proline rich C-terminal region of the adhesin related 30-kDa protein. The truncated adhesin related protein of 22 kDa showed reduced antigenicity compared to the corresponding wild-type protein. Moreover, the mutant M7 proved incapable of adhering to erythrocytes and to a human colon carcinoma cell line indicating that the repetitive C-terminal region of the 30 kDa protein is essential for effective cytadherence. The adhesin related 30-kDa protein as well as the truncated forms of the corresponding protein were accessible to carboxypeptidase Y which clearly shows surface exposure of the C terminus of this protein. PMID- 9228778 TI - Expression and phylogenetic relationships of a novel lacZ homologue from Actinobacillus pleuropneumoniae. AB - To learn more about the genetics and physiology of the important swine pathogen, Actinobacillus pleuropneumoniae, we cloned the lacZ gene by complementation of an Escherichia coli delta lac mutant. The A. pleuropneumoniae lacZ gene has an open reading frame of 3015 bp which could encode a protein with a predicted molecular mass of 117022. The deduced protein shares 26.8-34.8% identity with beta galactosidases from both Gram-positive and Gram-negative bacteria. Sequences with homology to seven regions commonly found in beta-galactosidases are present and amino acids corresponding to active site residues Tyr-503 and Glu-537 in E. coli LacZ are also conserved; however, there is a leucine in the place of Gly-794, a residue which has been implicated in substrate recognition. The sequences flanking the A. pleuropneumoniae lacZ gene do not share homology with known transport or regulatory genes nor do they share homology with cAMP receptor protein (CRP) or LacI binding sites. Low levels of beta-galactosidase activity could be detected when the protein was expressed from a multicopy plasmid in E. coli delta lac and when it was measured in A. pleuropneumoniae. The level of activity was not markedly reduced in the presence of glucose. Although the A. pleuropneumoniae LacZ shares some features with other beta-galactosidases, its constitutive expression and an unusual active site residue suggest that it may have a unique function. PMID- 9228777 TI - Delineation of the catalytic domain of Clostridium difficile toxin B-10463 to an enzymatically active N-terminal 467 amino acid fragment. AB - In an attempt to directly approach the postulated toxic domain of Clostridium difficile's TcdB-10463, eight subclones of different size and locations in the N terminal third of the toxin were generated. Expression of these toxin fragments was checked in Western blots and the enzymatic activity of the expressed proteins was analyzed by glucosylating Ras related small GTP-binding proteins. Two polypeptides of 875 aa (TcdBc1-3) and 557 aa (TcdBc1-H) glucosylated their targets Rho, Rac and Cdc42 with the same activity and specificity as the holotoxin. In comparison 516 aa (TcdBc1-N) and 467 aa (TcdBc1-A) protein fragments exhibited highly reduced activity, while Tcdc1 and TcdB2-3 (aa 1-243 and 244-890, respectively) were enzymatically inactive. Our results indicate that all structures involved in the catalysis are located at several different sites within the 557 aa fully active fragment. The shortest enzymatically still active protein covers aa 1-467 and obviously fulfils all minimal requirements for glucosylation. The data support the postulated three domain model of 'large clostridial cytotoxins'. PMID- 9228779 TI - 16S rDNA genotyping using PCR/RFLP (restriction fragment length polymorphism) analysis among the family Vibrionaceae. AB - The 16S rDNA genotypes among the family Vibrionaceae were determined using PCR/RFLP analysis. Five tetrameric restriction enzymes (HhaI, DdeI, RsaI, Sau3AI and MspI) were used for RFLP analysis and adequate numbers of informative bands were obtained from each enzyme. Twenty-seven genotypes were obtained from 49 type and reference strains including 35 species. Nineteen species could be assigned to specific 16S rDNA genotypes, supporting the application of this analysis for identification. Trees constructed using five endonucleases resolved groups almost identical to those inferred from 16S rRNA gene sequencing. However, the branch lengths and detailed relationships among strains within a group differed from those inferred from sequence comparisons. The results of this study should be useful for genotyping, identification and approximate classification of natural isolates belonging to the family Vibrionaceae. PMID- 9228780 TI - Bioavailability and biodegradation of polycyclic aromatic hydrocarbons in soils. AB - Inoculation of soil with bacteria (a Gram-negative rod [PD2] and a 4-membered consortium [DC1]) accelerated mineralisation of phenanthrene and pyrene (but not naphthalene) added individually to a pristine sand and a pristine organic soil. The half-life of naphthalene was 3.5 days in both soils whether inoculated or non inoculated. However, the half-life of phenanthrene decreased from 86 days in non inoculated sand soil and 80 days in the non-inoculated organic soil to 3.6 days in the sand and 3.1 days in organic soil when inoculated with PD2, and to 6.6 days in the sand and 8.7 days in the organic soil when inoculated with DC1. Phenanthrene mineralisation ceased after 23 days in DC1-inoculated soil and was 71.3 +/- 3.6% (sand) and 63.3 +/- 2.8% (organic). This compared with 96.8 +/- 3.8% (sand) 102.8 +/- 2.5% (organic) after 8 days in PD2-inoculated soil. Inoculation with DC1 (but not PD2) also accelerated mineralisation of pyrene, where the half-life decreased from 155 days to 18 days in the sand soil, and from 216 days to 33 days in organic soil. PMID- 9228781 TI - Characterisation of the peptidoglycan hydrolases of Listeria monocytogenes EGD. AB - The peptidoglycan hydrolase profile of Listeria monocytogenes EGD has been characterised under a variety of environmental and physiological conditions, using renaturing sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The profiles show activities ranging from 29 to 186 kDa. The 186-kDa enzyme was only observable under specific medium and aeration conditions. The enzyme activities show differential substrate specificity and sensitivity to incubation conditions. The peptidoglycan hydrolase profile of several different Listeria strains was also compared. PMID- 9228782 TI - Variation in the agr-dependent expression of alpha-toxin and protein A among clinical isolates of Staphylococcus aureus from patients with septicaemia. AB - In Staphylococcus aureus synthesis of many virulence factors is regulated by the agr locus. The regulatory molecule RNAIII, induced by agr, activates transcription of the alpha-toxin gene, hla, while it acts as a repressor of the protein A gene, spa. Forty clinical strains of S. aureus from human blood cultures were analysed for alpha-toxin and protein A production. An inverse correlation between alpha-toxin and protein A production was found in most strains. The levels of alpha-toxin and protein A production varied significantly among strains, which indicates various levels of the regulator, RNAIII. This was confirmed by selecting strains producing different amounts of alpha-toxin, showing that the variations in toxin production are due to the variations of RNAIII transcript. However, in one of the selected strains which produced high levels of alpha-toxin, OR153, although RNAIII is also strongly expressed, the specific hla mRNA was unexpectedly low. One partial explanation for the high alpha-toxin production by this clinical isolate might be its lack of extracellular proteases. PMID- 9228784 TI - Microbiological activity of whole and fractionated crude extracts of tea (Camellia sinensis), and of tea components. AB - Aqueous extracts of teas (Camellia sinensis) of different types and from various sources inhibited a wide range of pathogenic bacteria, including methicillin resistant Staphylococcus aureus. Tea extracts were bactericidal to staphylococci and Yersinia enterocolitica at well below 'cup of tea' concentrations. Activity was confined to one of four fractions obtained from a green tea extract by partition chromatography. Testing of pure tea compounds and closely related chemicals suggested that the antibacterial activity of extracts of green tea can be explained by its content of epigallocatechin, epigallocatechin gallate and epicatechin gallate. In black tea extracts, theaflavin and its gallates are additional antibacterially active components. PMID- 9228783 TI - Substitution of alanine for aspartate at position 179 in the SHV-6 extended spectrum beta-lactamase. AB - SHV-6 was previously identified by its susceptibility pattern and biochemical criteria in a clinical isolate of Klebsiella pneumoniae which was resistant to ceftazidime. It contains only a single point difference with the beta laSHV-1 gene as determined by PCR amplification and nucleotide sequencing. This is the result of a single amino acid substitution, Ala for Asp, at position 179. Directed mutagenesis experiments have shown this substitution to confer selective resistance to ceftazidime in the TEM family. PMID- 9228785 TI - Construction of a plasmid vector for transformation of Porphyromonas gingivalis. AB - A host-vector system for transformation of Porphyromonas gingivalis was constructed using a set (1) strains that can incorporate plasmid DNA by electroporation regardless of its source and (2) stable vector plasmids with a selectable marker. First, restriction-negative mutants were isolated, because P. gingivalis possesses restriction modification systems by which DNA introduced by transformation even from heterologous strains of the same species is excluded. For screening of the mutants, plasmid pE5-2 was employed since it could be transconjugated (mobilized) to P. gingivalis from Escherichia coli and is able to replicate in this species, albeit not stably. pE5-2 DNA prepared from E coli was introduced by electroporation into chemically mutagenized P. gingivalis cells. By this method, three putative restriction-negative clones were selected. These strains exhibited a capacity for electroporation with plasmid DNAs both from E. coli and from various P. gingivalis strains at a similar efficiency. Using one of the derivatives thus obtained, YH522, we then screened for plasmids that could replicate stably in P. gingivalis. Since no plasmids were found from P. gingivalis, cryptic plasmids from other species of black-pigmented oral anaerobic rods were examined for their ability to transform P. gingivalis. A series of plasmids constructed by ligation with pBR322 for replication in E. coli and the EcoRI-B fragment from pBF4 containing erythromycin resistance were prepared from E. coli and were used for electroporation of P. gingivalis. Among these, a recombinant plasmid containing the replicon of pYHBA1 from Porphyromonas asaccharolytica, designated pYH400, was found to be incorporated into the restriction-negative P. gingivalis strain and replicated stably. This set of recipient strains and stable plasmids with a selectable marker constitutes the first practical host-vector system for this species. PMID- 9228786 TI - Cloning and sequencing of the cDNA encoding lipase I from Trichosporon fermentans WU-C12. AB - A cDNA clone encoding extracellular lipase I (TFL I) from Trichosporon fermentans WU-C12 was isolated and characterized. The TFL I cDNA was isolated from a lambda gt10-based cDNA library using as a probe a 0.8 kb fragment, amplified by PCR with synthetic oligonucleotide corresponding to the partial amino acid sequences of TFL I. The cDNA encodes a protein consisting of 563 amino acids containing a putative signal peptide of 19 amino acids. The deduced amino acid sequence shares 99.5% overall identity with that of lipase II (GCL II) from Geotrichum candidum ATCC 34614, whereas TFL I is a trimer enzyme and GCL II monomer. Southern hybridization with the TFL I cDNA as a probe revealed that WU-C12 contained two different lipase genes. PMID- 9228787 TI - Isolation and characterization of a multiple peroxide resistant mutant from Xanthomonas campestris pv. phaseoli. AB - We have isolated a spontaneous multiple peroxide resistant Xanthamonas campestris pv. phaseoli mutant (XpHr). In the presence of peroxides, the mutant had a higher growth rate than the parent. It also had a greater than 100-fold increase in resistance levels to H2O2 killing but only slightly more resistance to tert-butyl hydroperoxide killing. Increases in activity were detected for the peroxide scavenging enzymes catalase (100-fold) and AhpC (over 30-fold). Also observed was cross-resistance to thermal killing; however, no cross-resistance to other oxidants or chemicals was found. Analysis of protein profiles revealed that proteins with molecular masses of 22 and 58 kDa were accumulated while proteins of 29, 33 and 41 kDa were depressed in the mutant. These results indicate that the mutant may have defect(s) in peroxide regulation, which resulted in high constitutive expression of peroxide scavenging enzymes. Nevertheless, the mutant retained growth phase dependent regulation of peroxide killing. The mutant should be useful in unravelling the nature of a complex peroxide stress regulon. PMID- 9228788 TI - Relationship between aflatoxin biosynthesis and sporulation in Aspergillus parasiticus. AB - Regulation of aflatoxin biosynthesis during differentiation of Aspergillus parasiticus was analyzed by using a drug that inhibits the development of the fungus and mutants affected in sporulation. Diaminobutanone, a competitive inhibitor of ornithine decarboxylase, repressed spore germination. If added after spore germination had occurred, it blocked sporulation completely and suppressed aflatoxin biosynthesis, but was only partially inhibitory of mycelial growth. Putrescine partially counteracted the inhibitory effect of the drug on both sporulation and aflatoxin biosynthesis. Analysis of mutants affected in sporulation confirmed the existence of a relationship between sporulation and aflatoxin formation. A nonsporulating mutant was unable to synthesize aflatoxins. In a sectorial mutant, the sporulating sector synthesized aflatoxins normally, whereas the asporogenous sector was unable to do so. It is suggested that regulation of aflatoxin biosynthesis is correlated with the sporulation process. PMID- 9228790 TI - Electrophoretic karyotype of clinical isolates of Paracoccidioides brasiliensis. AB - Chromosomal DNA molecules were obtained from eight clinical isolates of Paracoccidioides brasiliensis during the early staged of the yeast to mycelial transformation process and were separated by contour-clamped homogeneous electric field gel electrophoresis. In seven of the isolates, it was possible to separate five bands, albeit with two different migration patterns (A and B). In the remaining isolate only four bands were observed (pattern C). The isolated chromosomes showed molecular sizes ranging from 3.2 to 10 Mb according to Schizosaccharomyces pombe molecular size standards. The three different patterns observed had bands of 3.2, 3.8, 4.1, 5.2, 6.7, 7.2, 8.8, and 10 Mb. These data imply that at least 30 Mb of the genome are organized as chromosomal macromolecules. Due to the fact that P. brasiliensis spheroplasts are difficult to obtain, technical details are provided. PMID- 9228789 TI - Localization of cellobiose dehydrogenase in cellulose-grown cultures of Phanerochaete chrysosporium. AB - To elucidate the function of cellobiose dehydrogenase (CDH) in cellulose degradation by Phanerochaete chrysosporium, production and localization of CDH were investigated and compared with those in shaking and aerated static cultures grown on cellulose. Substantial CDH activity was detected in the medium of the shake cultures after 8 days of incubation, while no CDH activity was detected in the medium of static cultures at any point during the incubation period. Light microscopy clearly showed that many cellulose particles were adsorbed on the surface of the hypha in static cultures, whereas no cellulose particles were absorbed to the hypha is shake cultures. The addition of laminarinase to static cultures was very effective in detaching cellulose particles from the hypha surfaces. Using a potentiometric assay performed with an oxidation-reduction potential electrode, some CDH activity could be detected on the hypha/cellulose complexes in static cultures. Thus, CDH is produced also in static cultures, albeit in lower amounts that in shake cultures, but the enzyme is not released into the medium. It seem likely that the beta-1,3-glucan layer plays an important role in CDH localization and cellulose degradation. Immunocytochemical confocal laser scanning microscopy for the static cultures demonstrated that most CDH was adsorbed on the surface of the cellulose, especially around the cracks, which were formed by the action of cellulases during the course of incubation. From these observations, we conclude a direct participation of CDH in the degradation of cellulose in cooperation with cellulases. PMID- 9228791 TI - Characterization of mycobionts of photomorph pairs in the peltigerineae (lichenized ascomycetes) based on internal transcribed spacer sequences of the nuclear ribosomal DNA. AB - The "one fungus-two photomorphs" hypothesis suggests that certain lichenized fungi can establish a symbiotic relationship with either a eukaryotic or a prokaryotic photobiont. Such pairs of photomorphs are well know from cephalodiate Peltigerineae. Using an ascomycete-specific primer we amplified the internal transcribed spacer region of the nrDNA repeat of the mycobiont from total "lichen DNA" extracts of Peltigera malacea, photomorphs of P. aphthosa, P. britannica, and P. leucophlebia, Nephroma expallidum, and photomorphs of N. arcticum. Comparisons of 5.8S sequences suggest that the sequences obtained belong to the mycobiont and thus, that the ascomycete-specific primer is adequate for amplifying fungal DNA from total lichen-DNA extracts. The strict identity of nucleotide sequences of the internal transcribed spacer region of the nrDNA repeat between joined-photomorphs supports the one fungus-two photomorphs hypothesis. Photomorph may thus primarily reflect phenotypic plasticity of photomorphic fungi in response to changing environmental conditions. The cyanomorph recently reported for P. leucophlebia is shown to be based on a misidentified specimen of P. aphthosa. Comparisons of the ITS sequences further supports recognizing P. aphthosa, P. britannica, and P. leucophlebia at the species rather than the infraspecific level. PMID- 9228792 TI - Preparation and use of ion-exchange chromatographic supports based on perfluoropolymers. AB - A poly(vinyl alcohol) (PVA) coated particulate perfluoropolymer (FEP) support has been functionalised with ion-exchange groups for use in ion-exchange chromatography of proteins. Anion-exchange (DEAE and Q) and cation-exchange (SP) groups were introduced to PVA-FEP which had previously been activated using cyanuric chloride. The equilibrium adsorption capacities of SP-PVA-FEP were 31.8 and 25.2 mg ml-1 for lysozyme and IgG respectively while for DEAE-PVA-FEP, the equilibrium adsorption capacities were 14.9 and 9.7 mg ml-1 for beta lactoglobulin and HSA respectively. The equilibrium adsorption capacities for Q PVA-FEP were determined to be 17.2 and 13.5 mg ml-1 for beta-lactoglobulin and HSA respectively. Experiments carried out to investigate the resolving power of materials showed that both SP and Q-PVA-FEP were able to resolve proteins with only small differences in their isoelectric points and that this resolution could be maintained at a flow-rate of 1500 cm h-1. SP-PVA-FEP was used to purify lysozyme from egg whites where a 50-fold purification, to homogeneity, was achieved in 98% yield. The anion exchanger, Q-PVA-FEP could be used to purify G6PDH from a clarified homogenate of bakers' yeast 14.3-fold in 81% yield. PMID- 9228793 TI - Separation of enantiomers on a chiral stationary phase based on ovoglycoprotein. I. Influences of the pore size of base silica materials and bound protein amounts on chiral resolution. AB - The influences of the pore size of base silica gels and the bound amounts of ovoglycoprotein (OGCHI) on the chiral resolution of racemates have been investigated. To the 12-, 20- and 30-nm pore size silica gels, aminopropyl groups were introduced, activated with N,N'-disuccinimidyl carbonate and then reacted with OGCHI followed by blocking with D-glucosamine. For the OGCHI material prepared with the same pore size silica gel, a linear correlation was obtained between the capacity factor or each enantiomer and the amount of bound OGCHI. Enantioselectivity and resolution obtained with the reaction of 40 mg OGCHI per 1 g silica gel were lower than those obtained with the reaction of 80, 160 and 320 mg OGCHI, when the same pore size silica gel was used. This is due to the superfluous achiral interaction with base silica gels and/or spacers because of low protein coverage. With regard to comparison of the pore sizes of silica gel, the OGCHI materials prepared with the 12-nm pore size silica gel gave the largest capacity factor, and the highest enantioselectivity and/or resolution for the racemates tested, when the same amount of OGCHI was reacted. Thus, the OGCHI materials were prepared with the reaction of 80 mg OGCHI per 1 g silica gel having a 12-nm pore size followed by blocking with D-glucosamine. By diminishing the superfluous achiral interaction with base silica materials and/or spacers, much more efficient OGCHI materials should be obtainable. PMID- 9228794 TI - Reversed-phase separations of nitrogenous phospholipids on an octadecanoyl poly(vinyl alcohol) phase. AB - Molecular species of nitrogenous phospholipids (PLs) phosphatidylcholine (PC), phosphatidylethanolamine (PE), PE-derivatives and sphingomyelin (SP) were separated on an octadecanoyl poly(vinyl alcohol) (ODPVA) column by reversed-phase HPLC with UV and evaporative light scattering detection (ELSD). Mobile phases employed variable proportions of acetonitrile, methanol and water. HPLC-UV of the polar lipids yield components with peak intensities somewhat different from those obtained by HPLC-ELSD despite discernible similarity in the peak profiles observed in the two detection systems. Incorporation of ammonium hydroxide in mobile phases resulted in a decrease in analyte retention. The mobile phase basicity effect on capacity factors of PE species was significantly greater than that of PC counterparts. The new ODPVA-HPLC-ELSD technique was applied to the analysis of PC and PE molecular species in vegetable oils. PMID- 9228795 TI - Determination of phosphonates in natural waters by ion-pair high-performance liquid chromatography. AB - The paper describes a new method for the determination of phosphonates by ion pair high-performance liquid chromatography. The phosphonates are complexed with Fe(III) and separated on a reversed-phase polymer column. The eluent consists of a bicarbonate solution at pH 8.3, tetrabutylammoniumbromide as counter-ion and 14% acetonitrile. The complexes are detected at 260 nm. The four phosphonates HEDP (1-hydroxyethylene-1,1-diphosphonic acid). ATMP (aminotris [methylenephosphonic acid]), EDTMP [ethylenediaminetetra(methylenephosphonic acid)], and DTPMP [diethylenetriaminepenta (methylenephosphonic acid)] are well separated within 20 min. The limits of detection are 5.10(-8) M for ATMP and EDTMP, 1.10(-7) M for DTPMP and 5.10(-7) M for HEDP. The method is suitable for the determination of phosphonates in the influent and effluent of wastewater treatment plants. A preconcentration of 10 is easily achieved by adsorption of the phosphonates onto freshly precipitated calcium carbonate and subsequent dissolution of the solid-phase by HCl. PMID- 9228796 TI - Speciation of chromium dyes by high-performance liquid chromatography with inductively coupled plasma mass spectrometric detection. AB - High-performance liquid chromatography (HPLC) coupled to inductively coupled plasma mass spectrometry (ICP-MS) was employed for the separation and detection of chromium species in azo dyes, Acid blue 158 and Acid Blue 193; mainly Cr(III) and Cr(VI). The dyes were first analyzed for total metal content using ICP-MS and their stability in solution was studied by measuring their absorbance through a range of pH values. Then an isocratic chromatographic method employing reversed phase liquid chromatography with mass spectrometric detection was developed. Applying this method to the separation of these dyes, the absolute detection limits of Acid Blue 158 and 193 were 1 and 5 ng respectively. Additionally, Acid Blue 158 did not contain any chromium species. On the other hand, Acid Blue 193 contained uncomplexed and potentially bioavailable Cr(III). Acid Blue 193 did not have any toxic Cr(VI) present in the samples. PMID- 9228797 TI - Semi-micro solid-phase extraction of organic compounds from aqueous and biological samples. AB - A technique is described for performing solid-phase extractions on a semi-micro scale. Thin membrane disks 4 mm in diameter containing lightly sulfonated polystyrene or Silicalite particles are placed in the hub of a syringe needle. Aqueous samples (1-6 ml) are passed through the membrane disks and extracted compounds are subsequently eluted with 20-50 microliters of an organic solvent. Unlike solid-phase micro extraction (SPME) which uses a coated fiber, the present method is essentially a total extraction technique. Recoveries > 90% were generally obtained for a wide variety of test compounds. The same test compounds in human urine, albumin and human serum samples can be extracted without any pretreatment other than addition of a suitable surfactant. A "double-pass" technique was developed for convenient field sampling. PMID- 9228798 TI - Chromatographic analysis of volatile sulphur compounds in wines, using the static headspace technique with flame photometric detection. AB - This study describes the development of a method for determining eleven sulphur compounds in wine, which takes into account that thiols are easily oxidizable. The equilibria of the analytes between air and aqueous ethanol were studied and optimised using static headspace gas chromatography in order to obtain the best sensitivities. The influences of parameters such as temperature, time, ionic strength, headspace volume and the volume of headspace injected were determined. A cryogenic trap was used to concentrate the headspace analytes and they were chromatographically analysed using GC temperature programming on a poly(ethylene glycol) capillary column with FPD detection at 394 nm. The power relationship was observed between the chromatography response and a concentration of sulphur compounds in the range 2-150 micrograms l(-1) in the sample. Recoveries were determined by the standard addition technique and were higher than 90% for sulphides and disulphides and close to 80% for thiols. The overall method was successfully used to determine the sulphur compounds in white and red wines. PMID- 9228799 TI - Simultaneous quantitation of sixteen organochlorine pesticides in drinking waters using automated solid-phase extraction, high-volume injection, high-resolution gas chromatography. AB - A method is described for the simultaneous determination of sixteen organochlorine pesticides in drinking water using automated solid-phase extraction followed by high-volume (80 microliters) capillary column gas chromatography using electron capture detection. The fully automated extraction method followed by high-volume injection permits rapid sample analysis compared to previously described procedures since no further pre-concentration of the analytes is necessary after they have been eluted from the octadecyl solid-phase extraction cartridge. The lowest detectable concentrations of the pesticides are between 1-5 ng l(-1), relative recoveries range from 92-105% in tap water spiked at 100 ng l(-1) and the relative standard deviations are in the range 5-12%. PMID- 9228800 TI - Sensitive absorbance detection method for capillary electrophoresis based on laser wave-mixing. AB - Forward-scattering four-wave mixing is demonstrated as a sensitive absorbance detection method for capillary electrophoresis, using an argon ion laser operating at 457.9 nm. Since this four-wave mixing laser technique utilizes only two input laser beams, it offers important advantages, including ease of optical alignment, high wave-mixing efficiency and low excitation power requirements. In addition, since the analytical signal is a laser-like coherent beam, highly efficient optical signal detection can be performed with minimum optical background noise. Excellent detection sensitivity and short absorption path lengths, and hence, small detector probe volumes, are some of the useful features this absorbance detection method offers for on-column detection of both fluorescing and non-fluorescing analytes in capillary electrophoresis and liquid chromatography. Preliminary "detected" concentration detection limit of 8.5.10( 8) M, mass detection limit of 13 amol and an absorbance-unit detection limit of 1.35.10(-5) AU are determined for dabsyl-glycine using this absorbance detection method. PMID- 9228802 TI - Improved chiral separation using achiral modifiers in cyclodextrin modified capillary zone electrophoresis. AB - The influence of achiral modifiers on the chiral separation of propranolol is examined by cyclodextrin modified capillary zone electrophoresis. The improved chiral separation of propranolol is by molecules previously identified in our group as forming ternary complexes with cyclodextrin and pyrene. The polarity, chain size and heteroatom composition of the functional groups on the comodifiers was systematically varied in order to study the influence of these variables on the separation of propranolol. The improved chiral separation is accompanied by a decrease in retention time. The decrease in retention time is suggestive of a decrease in the association of beta-cyclodextrin (beta-CD) with propranolol which was verified by calculation of apparent association constants using fluorometric methods. PMID- 9228801 TI - Purification by reflux electrophoresis of whey proteins and of a recombinant protein expressed in Dictyostelium discoideum. AB - Protein purification that combines the use of molecular mass exclusion membranes with electrophoresis is particularly powerful as it uses properties inherent to both techniques. The use of membranes allows efficient processing and is easily scaled up, while electrophoresis permits high resolution separation under mild conditions. The Gradiflow apparatus combines these two technologies as it uses polyacrylamide membranes to influence electrokinetic separations. The reflux electrophoresis process consists of a series of cycles incorporating a forward phase and a reverse phase. The forward phase involves collection of a target protein that passes through a separation membrane before trailing proteins in the same solution. The forward phase is repeated following clearance of the membrane in the reverse phase by reversing the current. We have devised a strategy to establish optimal reflux separation parameters, where membranes are chosen for a particular operating range and protein transfer is monitored at different pH values. In addition, forward and reverse phase times are determined during this process. Two examples of the reflux method are described. In the first case, we described the purification strategy for proteins from a complex mixture which contains proteins of higher electrophoretic mobility than the target protein. This is a two-step procedure, where first proteins of higher mobility than the target protein are removed from the solution by a series of reflux cycles, so that the target protein remains as the leading fraction. In the second step the target protein is collected, as it has become the leading fraction of the remaining proteins. In the second example we report the development of a reflux strategy which allowed a rapid one-step preparative purification of a recombinant protein, expressed in Dictyostelium discoideum. These strategies demonstrate that the Gradiflow is amenable to a wide range of applications, as the protein of interest is not necessarily required to be the leading fraction in solution. PMID- 9228803 TI - Enantioseparation using selected polysaccharides as chiral buffer additives in capillary electrophoresis. AB - Selected water-soluble, native polysaccharides--such as amylose, laminaran, pullulan--and derivatized polysaccharides--methyl cellulose, hydroxypropyl cellulose, and carboxymethyl amylose sodium salt (CM-Am)--were investigated as chiral selectors in capillary electrophoresis. Effects of degree of polymerization and concentration of amylose on the separation of enantiomers of 1,1'-binaphthyl-2,2'-diyl hydrogen phosphate (BDHP) and a chiral cardiovascular drug cis-diltiazem were also studied. Pullulan and amyloses used in this study showed the same migration order for enantiomers of BDHP. In contrast, the migration order of BDHP enantiomers for cellulose derivatives and luminaran as well as with beta-cyclodextrin was opposite to that for amylose and pullulan. The enantioseparation of several chiral drugs was also performed using high-molecular mass amylose (Am-4900) and CM-Am. PMID- 9228804 TI - Interpretation of distortion product otoacoustic emission measurements. I. Two stimulus tones. AB - The interpretation of common but poorly understood observed characteristics of distortion product emissions is assisted by the development of a simple model. This model essentially includes only saturation of the cochlear amplifier, with emissions arising naturally from the same nonlinear processes which cause the saturation. The model provides useful physical explanations of emission behaviour, particularly considered as a function of the stimulus intensities of the two primaries, i.e., behavior with fixed stimulus frequencies. It is assumed that emission generation consists of two main components which are always present in the total emission, but which most often have approximately opposite, i.e., canceling, phases. One component arises in a small region centered about the peak of the emission generation function, while the other arises from the region basal to this peak. At low stimulus levels with normal cochlear amplifier operation, the peak of the emission generation function is sharp, so the emission from the peak region dominates the total emission. This "peak" emission has typically been characterized as the "active" emission. At high stimulus levels where saturation is important, or at all levels when the gain of the cochlear amplifier is reduced, the summed "basal" component dominates the total emission. The characteristics of this basal emission are similar to, and continuous with, the characteristics of the truly "passive" emission, i.e., the emission observed when the cochlear amplifier gain is identically zero. Under circumstances when the emissions from the peak and basal components are approximately equal, there is seen a sharp "notch" characteristic of phase cancellation. The simple model produces emission distributions as a function of independent variation of the two stimulus amplitudes which are in good agreement with observation. It is shown that the furosemide assay provides a good estimate of cochlear amplifier gain when a correction factor of about 10 dB is added. However, when using two stimulus tones, neither absolute emission amplitudes, or emission input-output functions, or the furosemide assay can adequately distinguish between cases of moderate versus poor cochlear amplifier dysfunction when the cochlear amplifier gains are in the range from about half normal to zero. PMID- 9228805 TI - Two-tone suppression of basilar membrane vibrations in the base of the guinea pig cochlea using "low-side" suppressors. AB - The responses of the basilar membrane (BM) in the basal section of the guinea pig cochlea were measured by laser interferometry. The stimuli were pairs of harmonically related tones, presented simultaneously. One tone, at the BM's characteristic frequency (CF) of about 17 kHz, was presented at a low intensity. The other tone, presented at various intensities, was a "low-side" suppressor, with a frequency of 0.2-8 kHz. As observed by many others, the suppressor tone, when presented at high enough intensity, reduced the magnitude of the CF component of BM displacement, sometimes dramatically. However, regardless of whether the CF component was suppressed or not, the sum of the displacement amplitudes of the CF and suppressor components was always greater than the displacement amplitude of the unsuppressed CF component. For suppressor frequencies up to 4 kHz, the suppression was both tonic and phasic, and synchronized to the suppressor period. For higher suppressor frequencies, principally tonic suppression was seen. PMID- 9228806 TI - Characterizing cochlear mechano-electric transduction in ears damaged with pure tones. AB - Cochlear microphonics were recorded in response to Gaussian noise from the round window of Mongolian gerbils. A nonlinear systems identification procedure provided the frequency-domain parameters of a third-order polynomial equation describing cochlear mechano-electric transduction (MET). Exposure to an 8 kHz pure tone at 100 dB SPL for 20 min reduced the magnitude of the linear, quadratic, and cubic terms significantly. Animals exposed to a 1- or 4-kHz pure tone showed changes in the quadratic term. Differentiation of the polynomial equation and algebraic manipulations of the coefficients provided physiologic indices of MET. The sensitivity, saturation voltages, and sound pressures required to saturate MET were altered in animals exposed to an 8-kHz pure tone. Limited changes occurred in animals exposed to a 1- or 4-kHz pure tone. PMID- 9228808 TI - Forward-masked intensity discrimination: duration effects and spectral effects. AB - Three experiments were completed to examine the effect of masker duration and spectrum on forward-masked intensity discrimination. Four listeners participated in each experiment. Intensity discrimination was measured in quiet and in the presence of forward maskers using adaptive forced-choice procedures. The standard duration was either short (10 ms) or long (250 ms) in experiment 1 and short (10 ms) in experiment 2. The standard always occurred 100 ms after the offset of the masker. In the first experiment employing 1.0-kHz maskers and standards, a short duration masker (10 ms) produced more masking than a long duration masker (250 ms). A mid-level elevation of the Weber fraction was observed for all conditions. To ensure that the results of experiment 1 were not influenced by off-frequency listening, the second experiment employed a broadband noise masker. As before, a short duration (10 ms) masker produced more masking than a long duration masker (100 ms) and a mid-level elevation of Weber fractions was observed. This outcome is inconsistent with a peripheral sensory effect for which an increase in masker duration should result in a greater amount of adaptation, and, as a consequence more masking. A third experiment employing a broadband noise masker and standard showed the greatest amount of masking for low-level standards, but only when the duration of the masker and standard was short. This result is similar to one seen for a single listener in the first experiment for short duration tonal maskers and standards. For this listener, a second tone presented at 4.133 kHz presented simultaneously with the 1.0 kHz masker reduced significantly the amount of masking for low-level standards, but the mid-level elevation of the Weber fraction remained. Taken together, these results suggest that perceptual similarity plays a role in forward-masked intensity discrimination but does not account entirely for the mid-level elevation of the Weber fraction. PMID- 9228809 TI - Effects of modulator phase for comodulation masking release and modulation detection interference. AB - In an effort to evaluate the importance of across-frequency comparisons of envelope patterns in comodulation masking release (CMR) experiments and to compare joint effects of target-masker frequency separation for both CMR and modulation detection interference (MDI) tasks, thresholds were measured for three tasks. These tasks were: (a) the detection of sinusoidal amplitude modulation (SAM) of a tone, (b) the detection of a reduction in the modulation depth of a fully modulated SAM tone, and (c) the detection of a tone added to a narrow band of noise. Thresholds were obtained for the target alone and for the target presented with two maskers. For the detection of SAM, thresholds did not depend on whether the modulation patterns of the target and masker elements were the same or random. For the latter two tasks, modulator phase effects were apparent for target-masker frequency separations less than 1-2 oct. In contrast, past work has shown that observers can compare modulator envelope phases across frequency separations larger than 1-2 oct [Strickland et al., J. Acoust. Soc. Am. 86, 2160 2166 (1989); Yost and Sheft, J. Acoust. Soc. Am. 85, 848-857 (1989)]. In a second experiment, thresholds for the detection of SAM were obtained after prolonged exposure to a fully modulated SAM tone. For four of the five observers, modulation-rate specific adaptation was obtained for test/adapting carrier frequency separations approaching 2 oct below and 1 oct above the adaptor. PMID- 9228807 TI - Absolute pitch and sex effect event-related potential activity for a melodic interval discrimination task. AB - Absolute pitch is a special ability which allows for special perceptual/cognitive strategies. Studies have shown differences in event-related scalp potentials between absolute-pitch (AP) and relative-pitch (RP) subjects of equal musical training. In this study, highly trained musicians (15 females/15 males) performed a melodic interval discrimination task, using intervals on-pitch in equal tempered tuning (A4 = 440 Hz) and tuned a half-semitone sharp. Subjects identified target intervals (probability 0.2) in a series of 400 randomly transposed intervals. AP subjects were expected to perform differently across intonation conditions, whereas RP subjects were not. Event-related potentials (ERPs) were recorded from three midline sites and two lateral sites. ERPs were analyzed by principal component analysis of variance. Sex was also considered as an independent subject variable. Performance was not significantly different either by absolute pitch or sex. Reaction times did not reveal any significant interactions involving AP or sex, but showed a significant effect by response type (target/nontarget). Strong P3 activity appeared to the target melodic intervals regardless of subject group or intonation. PCA factors with maxima at 352, 511, and 709 ms were sensitive to task relevance. Males showed greater positivity than females along the midline. A significant intonation by response type by sex interaction indicated a greater spread of values for females than males, and greater similarity in response by sex for the sharp than the on-pitch intervals. AP subjects showed reduced P3 activity along the midline, but increased over lateral sites. In a difficult musical task, the ERPs were sensitive to the sex of the listeners, as well as to whether they had absolute pitch. PMID- 9228810 TI - Profile analysis with an asynchronous target: evidence for auditory grouping. AB - Green and Dai [in Auditory Physiology and Perception, edited by Y. Cazals, L. Demany, and K. Horner (Pergamon, Oxford, 1992)] reported a series of experiments which suggested that listeners' ability to perform simultaneous across-frequency comparisons of intensity is severely impaired when the target and flanking components begin or end at different times. The present experiment sought to replicate the effect of onset asynchrony and included an additional condition in which the leading portion, of the asynchronous target component was accompanied by a pair of "captor" tones. The intended purpose of the captor tones was to promote a perceptual organization in which the leading and synchronous portions of the asynchronous target component were grouped with different auditory objects. For all six listeners an asynchrony of 320 ms raised thresholds substantially relative to the synchronous onset condition, the magnitude of the increase ranging between 6 and 16 dB. By contrast, the mean elevation of threshold in the presence of the captor was only 3 dB, although for all listeners thresholds were still greater than for the synchronous onset condition. The results support the view that the deleterious effect of onset asynchrony on profile analysis performance is due to the operation of auditory grouping principles. PMID- 9228811 TI - Discrimination of temporal asymmetry in cochlear implantees. AB - Several studies have recently demonstrated that normal-hearing listeners are sensitive to short-term temporal asymmetry in the envelopes of sinusoidal or noise carriers. This paper presents a study in which cochlear implantees were presented trains of current pulses with temporally asymmetric envelopes through one channel of an implant that stimulates the auditory nerve directly, thereby bypassing cochlear processes. When the level of the stimuli was adjusted to fit their audibility range, the implantees were able to discriminate temporal asymmetry over a much wider range than normal-hearing listeners. The results suggest that the perception of temporal asymmetry is limited by compression in the normal cochlea. PMID- 9228812 TI - Interference effects in short-term memory for timbre. AB - Four experiments investigated memory for timbre using the interpolated-tone paradigm [Deutsch, Science 168, 1604-1605 (1970)], in which participants discriminate pairs of tones (standard and comparison) separated by intervening (interpolated) tones. Interpolated tones varied from the standard tone in spectral similarity (within-dimensional variation), fundamental frequency (cross dimensional variation), and repetition frequency. While the latter two variables had negligible effects on timbre memory, interference with timbre memory increased with the spectral similarity of the interpolated tones to the standard tone. The findings closely parallel those found for pitch memory, and suggest that memory interference depends on perceptual similarity in both cases. PMID- 9228813 TI - Psychophysical studies with two binaural cochlear implant subjects. AB - Psychophysical studies have been completed with two binaural cochlear implant patients. In our earlier studies [van Hoesel et al., J. Acoust. Soc. Am. 94, 3187 3189 (1993); R. J. M. van Hoesel and G. M. Clark, Ann. Otol. Rhinol. Laryngol. Suppl. 106 104, 233-235 (1995)], lateralization experiments showed good sensitivity to interaural amplitudes but poor sensitivity to interaural time delays when compared with normal hearing subjects. In the studies presented here, both temporal and binaural intensity interactions were further explored. Interaural time delay (ITD) perception was investigated using direct measurement of the just-noticeable difference (jnd) in ITD. Both rate and place of stimulation were varied. Binaural rate discrimination was measured and compared with monaural rate perception. Binaural intensity interaction was explored for matched and unmatched place conditions by means of loudness summation and central masking studies. Results showed that ITDs for interaural time delays were large when compared to normal hearing, even when place of stimulation on each of the two sides was carefully matched. The jnds in ITD were similar for stimulation rates from 50 to 200 pps, and increased at 300 pps. Rate difference limens experiments showed similar results for diotic and monaural stimuli, but improved jnds for dichotic presentation at stimulation rates below 150-200 pps. Binaural intensity interactions showed loudness summation effects with both patients, for matched as well as unmatched place conditions. Central masking was also observed with both subjects, although it was not found to be place dependent. PMID- 9228814 TI - Performance over time of adult patients using the Ineraid or nucleus cochlear implant. AB - This study examined the average and individual performance over time of 49 adult cochlear implant subjects. Subjects were randomly assigned to receive either the Ineraid cochlear implant, with analog processing, or the Nucleus cochlear implant, with feature-extraction processing. All subjects had postlingual profound bilateral sensorineural hearing loss and received no significant benefit from hearing aids before implantation. Group data were examined in two ways. First, only subjects who had complete data over the test period were examined. Second, an analysis of all available data was carried out by mixed linear-model analysis. In this analysis, to account for missed follow-ups at the planned intervals, data consisting of the observations closest in time to the planned test times were modeled by natural splines with knots at the planned follow-up times. Contrasts between all pairs of planned follow-up times for each device were tested, as were contrasts between devices at each planned follow-up time. Results indicated little difference between the performance of the Ineraid and Nucleus subjects in their level of performance or their rate of learning. Postimplantation performance was typically superior to preimplantation performance within 9 months, and continued to improve up to 18-30 months depending on the speech perception measure. In some subjects, improvements in speech perception measures were observed up to four or five years postimplantation. There was also evidence that three subjects had a decrement in overall speech perception performance, although their postimplantation scores were always higher than their preimplantation scores. In at least one subjects this was likely a result of age-related cognition decrements. PMID- 9228815 TI - Articulatory control of phonological vowel length contrasts: kinematic analysis of labial gestures. AB - The present study investigated the articulatory control of the German vowel quantity contrast, i.e., the phonological difference between short and long vowels. By means of an optoelectronic system the excursions of the compound lower lip/jaw opening and closing gestures were measured during production of test sentences comprising the target sequence /pVp/ (V = /a/, /i/, /u/, /a:/, /i:/, /u:/). First, a highly linear relationship between peak velocity and movement amplitude emerged within each quantity class. Second, vowel quantity systematically influenced the scaling of velocity and amplitude during oral opening. Third, as compared to their short counterparts, long vowels showed an increased peakedness of the velocity profile. Finally, the velocity profiles of the long vowels were characterized by an asymmetric shape in terms of a prolonged deceleration phase of the opening and a lengthened acceleration interval of the closing movement. With respect to durational coarticulation patterns, the control of vowel quantity clearly differed from intrinsic, i.e., vowel type-induced, variability. The latter was partially compensated for the level of word duration by shortening of the prevocalic consonant whereas vowel quantity turned out to be primarily due to the lengthening of the syllable /pV/. PMID- 9228816 TI - Temporal and spectral estimations of harmonics-to-noise ratio in human voice signals. AB - The quantity, harmonic-to-noise ratio (HNR), has been used to estimate the level of noise in human voice signals. HNR estimation can be accomplished in two ways: (1) on a time-domain basis, in which HNR is computed directly from the acoustic waveform; and (2) on a frequency-domain basis, in which HNR is computed from a transformed representation of the waveform. An algorithm for computing HNR in the frequency domain was modified and tested in the work described here. The modifications were designed to reduce the influence of spectral leakage in the computation of harmonic energy, and to remove the necessity of spectral baseline shifting prescribed in one existing algorithm [G. de Krom, J. Speech Hear. Res. 36, 254-266 (1993)]. Frequency-domain estimations of HNR based on this existing algorithm and our modified algorithm were compared to time-domain estimations on synthetic signals and human pathological voice samples. Results indicated a highly significant, linear correlation between frequency- and time-domain estimations of HNR for our modified approach. PMID- 9228817 TI - Tongue surface displacement during bilabial stops. AB - The goals of this study were to characterize tongue surface displacement during production of bilabial stops and to refine current estimates of vocal-tract wall impedance using direct measurements of displacement in the vocal tract during closure. In addition, evidence was obtained to test the competing claims of passive and active enlargement of the vocal tract during voicing. Tongue displacement was measured and tongue compliance was estimated in four subjects during production of /aba/ and /apa/. All subjects showed more tongue displacement during /aba/ than during /apa/, even though peak intraoral pressure is lower for /aba/. In consequence, compliance estimates were much higher for /aba/, ranging from 5.1 to 8.5 x 10(-5) cm3/dyn. Compliance values for /apa/ ranged from 0.8 to 2.3 x 10(-5) cm3/dyn for the tongue body, and 0.52 x 10(-5) for the single tongue tip point that was measured. From combined analyses of tongue displacement and intraoral pressure waveforms for one subject, it was concluded that smaller tongue displacements for /p/ than for /b/ may be due to active stiffening of the tongue during /p/, or to intentional relaxation of tongue muscles during /b/ (in conjunction with active tongue displacement during /b/). PMID- 9228818 TI - Developmental weighting shifts for noise components of fricative-vowel syllables. AB - Previous studies have convincingly shown that the weight assigned to vocalic formant transitions in decisions of fricative identity for fricative-vowel syllables decreases with development. Although these same studies suggested a developmental increase in the weight assigned to the noise spectrum, the role of the aperiodic-noise portions of the signals in these fricative decisions have not been as well-studied. The purpose of these experiments was to examine more closely developmental shifts in the weight assigned to the aperiodic-noise components of the signals in decisions of syllable-initial fricative identity. Two experiments used noises varying along continua from a clear /s/ percept to a clear /[symbol: see text]/ percept. In experiment 1, these noises were created by combining /s/ and /[symbol: see text]/ noises produced by a human vocal tract at different amplitude ratios, a process that resulted in stimuli differing primarily in the amplitude of a relatively low-frequency (roughly 2.2-kHz) peak. In experiment 2, noises that varied only in the amplitude of a similar low frequency peak were created with a software synthesizer. Both experiments used synthetic /a/ and /u/ portions, and efforts were made to minimize possible contributions of vocalic formant transitions to fricative labeling. Children and adults labeled the resulting stimuli as /s/ vowel or /[symbol: see text]/ vowel. Combined results of the two experiments showed that children's responses were less influenced than those of adults by the amplitude of the low-frequency peak of fricative noises. PMID- 9228819 TI - Mechanisms of vowel recognition for Ineraid patients fit with continuous interleaved sampling processors. AB - Vowel recognition was assessed for eight, cochlear implant patients who use the Ineraid's six-electrode array. Recognition was tested in three conditions: with the Ineraid after years of experience; with a CIS processor at fitting of the processor; and with the CIS processor after 1 month's experience. At the time of fitting of the CIS processor, vowel recognition was not superior to that with the Ineraid. Recognition improved significantly over the period of a month. At 1 month, performance was significantly better with the CIS processor than with the Ineraid. This outcome is interpreted to mean that remapping of the vowel space is necessary following fitting with the CIS processor and some of the remapping occurs over a time period of days or weeks, rather than hours. Vowel errors at one month could be accounted for by two mechanisms. One is that patients attended to low-frequency channels at expense of high-frequency channels, or could not use information in high-frequency channels. The second is that, for diphthongs, patients could not detect frequency change over the course of the utterance. PMID- 9228820 TI - Perception of the American English liquid /ra-la/ contrast by humans and monkeys. AB - Human and monkey perception of the American English liquid /ra-la/ contrast was compared using various synthetic continua in which the normal spectral and temporal cues were higher either complete, partial, or altered in various ways. Two experiments compared human and monkey discrimination of the various continua using a low-uncertainty repeating-standard procedure. Results showed that, while human sensitivity was best at the human phoneme boundary, monkey sensitivity was best inside the /ra/ category. Also, while humans were more sensitive than monkeys to temporal variation in the stimuli, monkeys were more sensitive than humans to spectral variation, particularly for stimuli inside the /ra/ category. Two additional experiments compared human and monkey identification of the /ra la/ continua using a higher-uncertainty go/nogo identification procedure. Monkeys performed as accurately as humans (including one native Spanish and one native Hindi listener) in identifying the stimuli. However, human and monkey "phoneme boundaries" were in different places, with monkey boundaries shifted more toward /ra/ than human boundaries. These results suggest that human boundaries may be based on some sort of specific linguistic knowledge. Despite these boundary differences, monkeys showed a trading relation comparable to that of humans, indicating that the /ra-la/ trading relation has a psychoacoustic basis and that linguistic knowledge is not a necessary prerequisite for it to occur. PMID- 9228821 TI - Simulation of the effect of threshold elevation and loudness recruitment combined with reduced frequency selectivity on the intelligibility of speech in noise. AB - The effect of loudness recruitment and threshold elevation together with reduced frequency selectivity have been simulated to examine the combined effect of the two major consequences of cochlear hearing loss on the intelligibility of speech in speech-shaped noise. In experiment 1, four conditions were simulated: a moderate flat loss with auditory filters broadened by a factor of three (B3R2); a moderate-to-severe sloping loss with auditory filters broadened by a constant factor of three (B3RX); and these conditions with linear amplification applied prior to the simulation processing (B3R2+, B3RX+). For conditions B3R2 and B3RX, performance was markedly worse than for a control condition (normal hearing, condition R1) tested in a previous study. For conditions B3R2+ and B3RX+, linear amplification improved performance considerably. However, performance remained below that for condition R1 by between 5% and 19%. In experiment 2 the broadening of the auditory filters was made more realistic by making it a function of the absolute threshold at the center frequency of the auditory filter. Three different hearing losses were simulated: a moderate-to-severe sloping loss with variable broadening of the auditory filters (BXRX); the same moderate-to-severe sloping loss with linear amplification (BXRX+); and the same broadening of the auditory filters but without the simulation of loudness recruitment and threshold elevation (BX). For condition BXRX, performance was markedly worse than in condition R1, while performance in condition BX was somewhat worse than for condition R1. For condition BXRX+, linear amplification according to the NAL procedure improved performance to a large extent but it remained worse than for condition R1. The results are consistent with previous evidence indicating that only part of the decrease of performance produced by actual cochlear hearing loss can be compensated by conventional linear hearing aids. PMID- 9228822 TI - Dependence of ultrasonic attenuation and absorption in dog soft tissues on temperature and thermal dose. AB - The effect of temperature and thermal dose (equivalent minutes at 43 degrees C) on ultrasonic attenuation in fresh dog muscle, liver, and kidney in vitro, was studied over a temperature range from room temperature to 70 degrees C. The effect of temperature on ultrasonic absorption in muscle was also studied. The attenuation experiments were performed at 4.32 MHz, and the absorption experiments at 4 MHz. Attenuation and absorption increased at temperatures higher than 50 degrees C, and eventually reached a maximum at 65 degrees C. The rate of change of tissue attenuation as a function of temperature was between 0.239 and 0.291 Np m-1 MHz-1 degree C-1 over the temperature range 50-65 degrees C. A change in attenuation and absorption was observed at thermal doses of 100-1000 min, where a doubling of these loss coefficients was observed over that measured at 37 degrees C, presumably the result of changes in tissue composition. The maximum attenuation or absorption was reached at thermal dosages on the order of 10(7) min. It was found that the rate at which the thermal dose was applied (i.e., thermal dose per min) plays a very important role in the total attenuation absorption. Lower thermal dose rates resulted in larger attenuation coefficients. Estimation of temperature-dependent absorption using a bioheat equation based thermal model predicted the experimental temperature within 2 degrees C. PMID- 9228823 TI - Statistical properties of estimates of signal-to-noise ratio and number of scatterers per resolution cell. AB - Elementary theory underlying the relationship between the number of scatterers per resolution cell (N) and echo intensity signal-to-noise ratio (SNR) is reviewed. A relationship between the probability density functions for estimates of N and SNR2 is derived. This relationship is validated using a computer simulation. Phantom and in vitro experiments are described. In one set of experiments on phantoms, empirical distributions of estimates of N and SNR2 are measured and compared to theoretical predictions. The utility of SNR2 for discrimination of phantoms with different values for N is assessed using receiver operating characteristic (ROC) analysis. In another set of experiments, the frequency dependence of the SNR2 estimate is investigated for a two-component phantom and for excised dog kidney. It is shown that the frequency dependence of the SNR can help to identify the presence of two or more scattering components that are spatially mixed. With regard to kidney data, measurements performed both parallel and perpendicular to the predominant nephron orientation are reported. The observed anisotropy is compared to the anisotropy of backscatter coefficient encountered in previous investigations. PMID- 9228824 TI - Using acoustic radiation force as a concentration method for erythrocytes. AB - We investigated the potential damage inflicted on erythrocytes by acoustic radiation force when the cells are concentrated by a 500-kHz ultrasonic standing wave at the pressure node. The extent of the damage was estimated from the concentrations of potassium ions, iron complexes, and lactate dehydrogenase released from the cells. After 2 min of ultrasound irradiation at 12.8 mJ/m3, the cells concentrated on the pressure node, with a cell distribution half-width of 138 microns; no significant release of intracellular components was detected, even after 15 min of irradiation. The results indicate that even small ions like potassium are not released as a result of ultrasound irradiation on cell membranes without cavitation, and they demonstrate the potential use of acoustic radiation force for concentrating living cells in biomedical applications. PMID- 9228825 TI - Fractal dimension of sustained vowel productions in neurological dysphonias: an acoustic and electroglottographic analysis. AB - In order to investigate whether nonlinear methods of signal analysis provide a measure of phonatory irregularities in neurogenic voice disorders, the present study computed the fractal dimension (D) both of the electroglottographic (EGG) and the acoustic signal of sustained vowel productions obtained from patients with Parkinson's disease (PD) and cerebellar atrophy (CA). Compared with normal speakers, the female PD group as well as the male and female CA patients showed an increased dimension (D) of the EGG. The dimensional complexity of the acoustic signal largely depended on vowel type. Furthermore, the dimension of the acoustic signal was reduced in male PD patients as compared to the respective controls. PMID- 9228826 TI - [The role of surgery in the treatment of appendicular abscesses]. AB - The management of appendiceal abscesses is still discussed and many different approaches are nowadays adopted. The aim of this study was to analyze retrospectively our experience with this disease to value the results of drainage of the abscess and appendectomy in one stage in presence of appendiceal abscesses. We studied 44 patients consecutively observed in our Department of General Surgery all submitted to drainage of the abscess and appendectomy for acute appendicitis with periappendiceal abscess. Preoperative ultrasonography showed an accuracy of 85.7% in detecting the presence of an abscess. Mean size of the abscesses were 5 cm (from a minimum of 3 cm to a maximum of 9 cm). The mean duration of surgical operation was 48 minutes (min 35'-max 95'), with a mean in hospital stay of 6.2 days. Morbidity rate was 9% and was due in 75% of cases to wound infection and in 25% of cases to wound dehiscence. Neither major morbidity nor mortality were observed. In consideration of the results the authors conclude that even in presence of an appendiceal abscess, appendectomy with abscess drainage is not only a safe operation with a low morbidity rate but the procedure of choice allowing a significative reduction of hospitalization and health cost. PMID- 9228827 TI - Peptide growth factors: clinical and therapeutic strategies. AB - The literature contains many accounts of studies in which tumour growth has been accelerated by administration of a particular mitogen and the response then inhibited by co-administration of the corresponding antagonist. Much effort has been focused on the development of cytokine or growth factor antagonists. Like most other cancer therapies, biological therapies will undoubtedly have undesirable toxicities because the proteins they target may not be unique to malignant cells. We reviewed the clinical and therapeutic potential of growth factor agonists and antagonists in some non urologic and urologic diseases. In a recent report we demonstrated that both androgen and antiandrogen treatments enhance the proliferation rate of the hormone-dependent prostate cancer cell line LNCaP, expressing a mutated androgen receptor. Simultaneous treatment with 1 nM R1881 and 100 nM OH-Flutamide, completely counteracted the androgen-induced increase of Epidermal Growth Factor (EGF) levels. Moreover we found that Testosterone, DHT and EGF are mainly concentrated in the periurethral zone in human BPH and long term treatment with Finasteride and with Flutamide modify the distribution and concentration of these factors. Some authors analyzed whether and addition of aurin tricarboxylic acid (ATA) can reduce the growth rate of basic FGF-dependent cells in a manner similar to suramin. PMID- 9228828 TI - Ureteral injuries during gynecologic procedures. AB - The reported incidence of iatrogenic ureteral injuries ranges from 0.05 to 30%. These injuries are particularly secondary to gynecologic surgical procedures. Simple abdominal hysterectomy has proved to be the most common procedure leading to such injures. Because some ureteral injuries may be symptomless, leading to silent kidney loss, the incidence may be too low, and a figure of up to 2.5% after gynecologic operations has been suggested. The incidence continues to be about ten cases during abdominal surgery for one case during vaginal surgery. Different risk factors may influence the ureteral injuries rate. An operation at the pelvic brim, distorted anatomy, removal of the adnexa or of ovarian neoplasm, may facilitate the occurrence of an ureteral trauma. The management of ureteral trauma is positively influenced by an early recognition of the trauma. Conventional technique or ureteroneocystostomy or end to end anastomosis with ureteral stent to treat the injury proved successful. Extensive reconstruction draws upon the entire therapeutic armamentarium of the urologist. Surgical options mainly include creation of bladder tubes and autotransplantation. The most important factor influencing the management of ureteral injury is the presence of associated complications. Blandy et al. sustained to attempt repair of these complicated iatrogenic injuries as soon the diagnosis has been made. PMID- 9228829 TI - Homophobia and heterosexism in social workers. AB - Evidence suggests that social workers may be biased when dealing with gay and lesbian populations. The study discussed in this article attempted to measure the extent of homophobia and heterosexist bias and their correlates in a cohort of 187 social workers using the Index of Attitudes toward Homosexuality, the Attitudes toward Lesbians and Gay Men Scales, and a newly created scale to measure heterosexist bias. We found that 10 percent of respondents were homophobic and that a majority were heterosexist. Levels of homophobia and heterosexism were negatively correlated with amount of social contact with homosexual men and women. Religiosity was associated with higher levels of homophobia and heterosexism, and having been in psychotherapy was associated with more positive attitudes toward gay men and lesbians. Amount of education on topics related to homosexuality was not correlated with levels of homophobia and heterosexism. PMID- 9228830 TI - Factors associated with early sexual activity among urban adolescents. AB - This study uses lifespan and ecological frameworks to investigate the factors associated with early adolescent sexual activity. Data from a longitudinal study of urban teenagers of color address three issues: (1) the prevalence and pattern of sexual activity among boys and girls ages 15 and younger, (2) the link between early sexual activity and high-risk sexual behavior, and (3) the life contexts linked with early sexual activity. Results from 803 African American and Hispanic adolescents suggest a high prevalence of early sexual activity, which is associated with higher rates of childbearing and risky sexual behavior than sexual activity initiated in later adolescence. Somewhat different factors are associated with early sexual activity for boys and girls, although family composition, parent attachment, and substance use are important for both genders. Implications for intervention are discussed. PMID- 9228831 TI - Social support and depression among older adults living alone: the importance of friends within and outside of a retirement community. AB - Using socioemotional selectivity theory as a framework, the study described in this article examined the extent to which social support from friends both within and outside of a retirement community was associated with depression. Although levels of social support from friends within the retirement community were quantitatively high, they failed to have a significant effect on depression. In contrast, social support from friends living elsewhere consistently predicted low levels of depression. Practice implications include the importance of maintaining friendship ties with people living elsewhere and of strengthening friendship ties within the retirement community. PMID- 9228832 TI - Case advocacy in child welfare. AB - This article examines the concept of advocacy in child welfare. Definitions of social advocacy and case advocacy are presented that will perhaps give child advocates, child welfare workers, and social workers a better understanding of the concepts and their roles in the protection of children who have been abused or neglected. A historical account of advocacy efforts on behalf of abused and neglected children in the United States is presented, followed by an analysis of current advocacy efforts, highlighting the potential of court-appointed special advocates (CASAs). The article also presents implications and ideas for social workers to assist in case advocacy efforts for children who are part of the child welfare system. PMID- 9228833 TI - Gnathologist's definition of centric relation. PMID- 9228834 TI - Impacted maxillary central incisor in mixed dentition treatment. AB - The impacted maxillary central incisor in a child poses a disturbing esthetic dilemma to parents, by virtue of its location. Neither the orthodontist nor the parents prefer to wait for the fully erupted permanent dentition and comprehensive orthodontics to resolve this problem. This challenge can be met in the early mixed dentition stage, as described with these two cases. The response to the technique was excellent. The patients now possess confidence to smile and have enhanced self-esteem, which is critical even in early life. Just as important is that the newly positioned maxillary central incisor is the natural tooth, not a temporary prosthesis that can loosen or break. Gingival contour is a consideration, however. PMID- 9228835 TI - The functional matrix hypothesis revisited. 1. The role of mechanotransduction. AB - The periodic incorporation of advances in the biomedical, bioengineering, and computer sciences allow the creation of increasingly more comprehensive revisions of the functional matrix hypothesis. Inclusion of two topics, (1) the mechanisms of cellular mechanotransduction, and (2) biologic network theory, permit this latest revision; presented here in two interrelated articles. In this first article, the several possible types of intracellular processes of mechanotransduction are described. These translate the informational content of a periosteal functional matrix stimulus into a skeletal unit (bone) cell signal. The correlation between the strengths of the endogenous electrical fields produced by muscle skeletal muscle activity, and those to which bone cells maximally respond are stressed. Further, a physical chain of macromolecular levers, connecting the extracellular matrix to the bone cell genome is described, suggesting another means of epigenetic regulation of the bone cell genome, including its phenotypic expression. PMID- 9228836 TI - T-loop position and anchorage control. AB - The effect of off-center positioning on the force system produced by segmented 0.017 x 0.025-inch TMA T-loops was measured. A T-loop was designed to produce equal and opposite moments in the centered position. The spring was tested in seven positions, centered, 1, 2, and 3 mm toward the anterior attachment, and 1, 2, and 3 mm toward the posterior attachments. The horizontal force, vertical force, and alpha and beta moments were measured over 6 mm of spring activation. The results showed that the alpha/beta moment ratio was dependent only on the spring position, and independent of spring activation. Eccentric positioning of T loop springs effectively produces a consistent moment differential through the range of spring activation. PMID- 9228837 TI - Initial effects of treatment of Class II malocclusion with the Herren activator, activator-headgear combination, and Jasper Jumper. AB - The initial effects of treatment of Class II, Division 1 malocclusion with an activator, according to Herren (27 patients), with an activator-headgear combination (20 patients), or with the Jasper Jumper appliance (25 patients) were studied on lateral cephalograms from before and after 6 to 8 months of treatment. The patients' ages ranged from 9 to 12 years. At the end of the period of observation, the correction in overjet and molar relationship was more complete in the patients with the Jasper Jumper than in the patients with the activator. Whereas all the patients with the Jasper Jumper showed neutral occlusion, this was the case in only 20 of the 47 patients with the activator. The correction of the distal occlusion occurred through a combination of skeletal and dentoalveolar adaptations. Skeletal changes accounted for 42%, 35%, and 48% of the overjet correction by the Herren-type activator, the headgear-activator, and the Jasper Jumper, respectively. The correction of the molar relationship occurred to 55%, 46%, and 38% by skeletal changes in the respective groups. Dentoalveolar compensation (distal movement of the upper molars, mesial movement of the lower molars) appeared to be inversely related to skeletal adaptation. The patients with the Jasper Jumper showed a marked intrusion of the lower incisors with a consequent reduction in overbite. PMID- 9228838 TI - A size-standardized analysis of soft tissue facial profile during growth. AB - A method for the quantitative and qualitative analysis of the facial soft tissue profile has been developed and applied to analyze the age differences in lateral cephalograms for the annual Bolton standards from ages 1 to 18 years. To standardize for different facial sizes, profiles were traced in polar coordinates without modifications of facial shape. Most of the soft tissue landmarks showed progressive modifications from birth to 18 years of age. Soft tissue nasion and lower lip had a steep change between 2 and 3 years of life, pronasale between 3 and 5 years of life, A', upper lip, and stomion between 4 and 5 years of life. Hereafter, all these landmarks but N' had several minor modifications progressing toward the adult value. Soft tissue nasion did not modify significantly after 2 years of age. Conversely, changes in the relative positions of B' and Pg' were more scattered in the analyzed period. Age-related size differences were more linear than shape modifications, with gradual increments from 1 to 18 years of age. The method allowed a simple and rapid quantitative evaluation of soft tissue profiles during facial growth. An approximate evaluation of the soft tissue thickness at nose, lips, and chin was also possible. No particular mathematical knowledge was required at any step of the analysis. Results were in good agreement with the well-known patterns of normal growth and development, thus confirming the practical possibilities of the method. PMID- 9228839 TI - Clinical characteristics and properties of ceramic brackets: A comprehensive review. AB - The aim of this article is to provide the clinician with an up-to-date comprehensive literature review concerning the clinical characteristics and properties of ceramic brackets. The article critically presents and discusses the various aspects of ceramic brackets with regard to the types, physical properties, bond strength, frictional resistance, base surface characteristics, debonding techniques and enamel fracture risks, enamel abrasion and wear, bracket fracture, and bracket recycling. The article contains also guidelines and criteria concerning the specific applications and use of ceramic brackets. PMID- 9228840 TI - The effect of friction on the bending stiffness of orthodontic beams: a theoretical and in vitro study. AB - The purpose of this study was to investigate the effect of friction on the bending stiffness of orthodontic beams. A theoretical and experimental model have been established where tensile and compressive forces are applied to an arch wire to simulate the effect of additional friction during activation and deactivation, respectively. The results show that tensile force increases wire stiffness, and that compressive force increases flexibility. Thus more force will be needed during activation and more force will be lost during deactivation. The amount of force lost increases nearly linearly with increasing friction. During activation, the percentage increase in force due to friction for a given deflection is about equal to the loss of force due to friction during deactivation. Friction affects thin flexible wires more than heavy wires. Careful ligation is recommended in the leveling phase to reduce the negative side effects of friction. PMID- 9228841 TI - Normal faciolingual inclinations of tooth crowns compared with treatment groups of standard and pretorqued brackets. AB - Faciolingual inclinations of tooth crowns were measured on plaster casts of normal occlusions and posttreatment orthodontic models of treated cases. Normal anatomic occlusion, standard edgewise and Roth bracket groups of 10 subjects were examined for a change in torque values. On study models, crown inclinations of right and left central teeth to second molar in the upper and lower dental arches were measured to the functional occlusal plane and mean tooth inclinations were calculated. In normal occlusion group, upper central and lateral teeth inclined lingually, lower central teeth inclined slightly labially and lower lateral teeth inclined lingually but the standard deviations of mean values for upper and lower anterior teeth were high. Upper posterior teeth, from canine to molar, had a lingual inclination and lower posterior teeth had a progressively increasing lingual inclination from lateral to molar teeth. In treatment groups, upper central and lateral teeth had labial crown inclination and lower molar teeth had more lingual inclination as compared with the normal occlusion group. No significant variation was found between the mean torque values of standard and Roth treatment groups. PMID- 9228842 TI - Oral environment temperature changes induced by cold/hot liquid intake. AB - The use of NiTi shape memory alloys, introduced into orthodontics because of their ability to develop light continuous forces that prove more effective than heavy intermittent forces in the teeth movement, requires the mastering of the functional properties of NiTi wires. More specifically, the recovery force acting on the teeth is a sensitive function of temperature: knowledge of oral temperature modifications is therefore required to understand the stress state modification felt during orthodontic therapy. The temperature modifications induced by cold or hot drink intake in the oral cavity were investigated by using arch wires, fixed to removable Hawley retainers, similar to those currently used in orthodontic practice, by means of six temperature sensors placed in correspondence with specific teeth. Similarly, the temperature changes were detected on a metallic frame, fixed onto the palatal zone to a Hawley retainer, where a palatal expander was placed to correct unilateral or bilateral crossbites in deciduous or in early mixed dentition. The maximum temperature change was observed in the interincisor area: The temperature modification on other teeth depends on the modality of drink intake, with the highest temperature variations being detected in the palatal zone. Hence modifications in the stress state during orthodontic therapy with NiTi wires are to be expected. PMID- 9228843 TI - The effects of ion implantation on rate of tooth movement: an in vitro model. AB - Recently, the ion-implantation process has been applied to orthodontic wires. By altering the surface composition of a wire, the ion-implantation process supposedly decreases the frictional forces produced during tooth movement. The purpose of this study was to compare the amount of tooth movement produced by different orthodontic wire compositions, under identical conditions, by using an in vitro model. The wires tested were stainless steel, nickel-titanium (control and ion implanted), and beta-titanium (control and ion implanted). The amount of tooth movement was measured and compared. Results demonstrate that, stainless steel produced the least frictional force during in vitro tooth movement, followed by ion-implanted nickel-titanium, ion-implanted beta-titanium, untreated nickel-titanium, and finally, untreated beta-titanium. A Wilcoxon rank sum test showed statistically significant differences in the amount of movement seen with the ion-implanted wires when compared with their untreated counterparts. PMID- 9228844 TI - Corrosion behavior of 2205 duplex stainless steel. AB - The corrosion of 2205 duplex stainless steel was compared with that of AISI type 316L stainless steel. The 2205 stainless steel is a potential orthodontic bracket material with low nickel content (4 to 6 wt%), whereas the 316L stainless steel (nickel content: 10 to 14 wt%) is a currently used bracket material. Both stainless steels were subjected to electrochemical and immersion (crevice) corrosion tests in 37 degrees C, 0.9 wt% sodium chloride solution. Electrochemical testing indicates that 2205 has a longer passivation range than 316L. The corrosion rate of 2205 was 0.416 MPY (milli-inch per year), whereas 316L exhibited 0.647 MPY. When 2205 was coupled to 316L with equal surface area ratio, the corrosion rate of 2205 reduced to 0.260 MPY, indicating that 316L stainless steel behaved like a sacrificial anode. When 316L is coupled with NiTi, TMA, or stainless steel arch wire and was subjected to the immersion corrosion test, it was found that 316L suffered from crevice corrosion. On the other hand, 2205 stainless steel did not show any localized crevice corrosion, although the surface of 2205 was covered with corrosion products, formed when coupled to NiTi and stainless steel wires. This study indicates that considering corrosion resistance, 2205 duplex stainless steel is an improved alternative to 316L for orthodontic bracket fabrication when used in conjunction with titanium, its alloys, or stainless steel arch wires. PMID- 9228845 TI - Predictive variables for the outcome of early functional treatment of Class III malocclusion. AB - The aim of this study was to select a model of cephalometric and occlusal predictive variables for the results of early treatment of Class III malocclusion, due to mandibular protrusion in the deciduous dentition. Lateral cephalograms and dental casts of 45 subjects (22 boys and 23 girls) with Class III malocclusion in the deciduous dentition were analyzed at the start of treatment (mean age 5.57 +/- 0.85 years). The patients were treated with a functional appliance (removable mandibular retractor). All subjects were reevaluated after a mean period of 9.54 +/- 2.28 years comprising active treatment plus retention. At this time, the sample was divided into two groups according to occlusal criteria: successful group (23 subjects) and unsuccessful group (22 subjects). Stepwise variable selection on the measurements at the time of first observation identified three predictive variables; the inclination of the condylar axis in relation to the stable basicranial line (CondAx-SBL), the inclination of the nasal line to the mandibular line (NL-ML), and the transverse width of the mandibular arch measured at the first deciduous molars. Discriminant analysis assigned a classificative power of 95.55% to the predictive model. On the basis of the equation generated by the multivariate statistical method, outcome of early treatment for each new case with Class III malocclusion in the deciduous dentition can be accordingly predicted. The important role of both vertical and transverse relationships in Class III prognosis was emphasized. PMID- 9228846 TI - The mechanism of Class II correction in late Herbst treatment. AB - The aim of this study was to analyze quantitatively the sagittal skeletal and dental changes contributing to Class II correction in patients treated with the Herbst appliance after the pubertal growth peak. The sample consisted of 21 subjects with a Class II, Division 1 malocclusion treated during the skeletal maturity stages MP3-H and -1, corresponding to a period after the maximum of pubertal growth (late treatment). A comparison was made with 22 Herbst subjects treated during the skeletal maturity stages MP3-E and -F, corresponding to a period before the maximum of pubertal growth (early treatment). Lateral head films from before and after Herbst therapy were analyzed, according to the method of Pancherz. As a result of the Herbst therapy, all patients attained a Class I or overcorrected Class I occlusal relationship. Class II molar correction averaging 6.1 mm was due to 37% skeletal and 63% dental changes. Overjet correction averaging 8.4 mm was due to 27% skeletal and 73% dental changes. Differences between the late and the early treated patients were only found for the dental changes. The upper anterior teeth were retroclined and the lower anterior teeth were proclined more in the late cases. The conclusion of the study was that the Herbst appliance is equally efficient in patients treated before and after the pubertal peak of growth. However, proclination of the lower incisors (anchorage loss) in late treated subjects is larger than in early treated subjects. This should be considered in treatment planning. PMID- 9228847 TI - Long-term effects of Herbst treatment on the mandibular incisor segment: a cephalometric and biometric investigation. AB - The purpose of this study was to analyze mandibular incisor changes during and after Herbst treatment with respect to tooth inclination and anterior crowding. The sample consisted of 24 Class II, Division 1 subjects (15 boys and 9 girls) treated with Herbst appliance. Dental casts and lateral head films from before and after treatment, 6 months after treatment and at the end of the growth period (at least 5 years after treatment) were analyzed. During treatment, the lower incisors were proclined (ILi/ML) an average of 10.8 degrees and the incisal edge (li) moved anteriorly by 3.2 mm. The available space and the irregularity index in the lower anterior region were in general unaffected by therapy. During the first posttreatment period of 6 months, the lower incisor inclination (ILi/ML) recovered an average of 7.9 degrees and the incisal edge (li) moved posteriorly by 2.5 mm. However, the available space was almost unchanged. During the second posttreatment period, i.e., from 6 months after treatment to the end of growth, the lower incisor inclination remained on average unchanged in relation to the mandibular plane (ILi/ML) but the teeth retroclined in relation to the nasion sella line (ILi/NSL). The available space decreased (mean 0.8 mm, p < 0.01) and the irregularity index increased (mean 2.0 mm, p < 0.01). The correlation between changes in the ILi/NSL and in the NSL/ML angles was moderate (r = -0.57, p < 0.01), indicating that the reduction in the ILi/NSL angle was partly a result of anterior mandibular growth rotational changes. In conclusion, it can be said that the proclination of the lower front teeth during Herbst treatment did not result in incisor crowding after treatment. In a long-term perspective, the development of incisor crowding was thought to be associated with normal craniofacial growth changes. PMID- 9228848 TI - The long-term stability of LeFort I maxillary downgrafts with rigid fixation to correct vertical maxillary deficiency. AB - This study evaluated the long-term stability of LeFort I maxillary downgrafts with rigid fixation in 28 patients with vertical maxillary deficiency. Preorthodontic, presurgical, immediate postsurgical, and an average of 16 months postsurgical cephalometric radiographs were evaluated. The results indicated that overall vertical maxillary stability after downgraft was good with 80% of the cases showing superior relapse of 2 mm or less. The mean amount of postoperative relapse represented 28% of the surgical downgraft. No correlation was found between the amount of the maxillary downgraft and the subsequent postsurgical vertical stability of the maxilla. No difference was found in the vertical stability of the maxilla comparing one-jaw and two-jaw procedures. It was also found that there was no difference in the vertical stability of the maxilla between single-segment and multiple-segment maxillary procedures, and also when comparing standard and high LeFort I osteotomy techniques. In addition, occlusal plane rotation of surgery, as well as the type of presurgical orthodontic movement, were both found to have no influence on postoperative stability of the maxilla. PMID- 9228849 TI - Ideal digital imaging system: a preliminary report. PMID- 9228850 TI - Two versus three years? PMID- 9228851 TI - Reinke's edema: phonatory mechanisms and management strategies. AB - Reinke's edema (RE) has been associated typically with smoking and sometimes with vocal abuse, but aspects of the pathophysiology of RE remain unclear. To gain new insights into phonatory mechanisms associated with RE pathophysiology, we used an integrated battery of objective vocal function tests to analyze 20 patients (19 women) who underwent phonomicrosurgical resection. Preoperative stroboscopic examinations demonstrated that the superficial lamina propria is distended primarily on the superior vocal fold surface. Acoustically, these individuals have an abnormally low average speaking fundamental frequency (123 Hz), and they generate abnormally high average subglottal pressures (9.7 cm H2O). The presence of elevated aerodynamic driving pressures reflects difficulties in producing vocal fold vibration that are most likely the result of mass loading associated with RE, and possibly vocal hyperfunction. Furthermore, it is hypothesized that in the environment of chronic glottal mucositis secondary to smoking and reflux, the cephalad force on the vocal folds by the subglottal driving pressure contributes to the superior distention of the superficial lamina propria. Surgical reduction of the volume of the superficial lamina propria resulted in a significant elevation in fundamental frequency (154 Hz) and improvement in perturbation measures. In almost all instances, both the clinician and the patient perceived the voice as improved. However, these patients continued to generate elevated subglottal pressure (probably a sign of persistent hyperfunction) that was accompanied by visually observed supraglottal strain despite the normal-sized vocal folds. This finding suggests that persistent hyperfunctional vocal behaviors may contribute to postsurgical RE recurrence if therapeutic strategies are not instituted to modify such behavior. PMID- 9228852 TI - Vocal fold atrophy: quantitative glottic measurement and vocal function. AB - Videostroboscopic glottic measurements and vocal function were evaluated in 41 vocal fold atrophy patients with bowed vocal folds. The amount of bowing in the resting position and the glottal gap area and vibratory amplitude during phonation were measured from digitized videostroboscopic images. Vibratory amplitude was not decreased on atrophic vocal folds. With the same amount of total bowing, the glottal gap area for bilateral atrophy was smaller than for unilateral atrophy. These results suggest that vocal fold atrophy is not disadvantageous to thyroplasty type I, and that bilateral procedures may produce a better outcome than a unilateral procedure in the treatment of bilateral atrophy. Acoustic, aerodynamic, and perceptual parameters of vocal function were measured. The acoustic high-frequency power ratio and the H-index correlated with the glottal gap area. The mean flow rate correlated with the amount of bowing. The degree of dysphonia was related to the size of the glottal gap and bowing. PMID- 9228853 TI - Changes in length and spatial orientation of the vocal fold with arytenoid adduction in cadaver larynges. AB - Videoendoscopy suggests that arytenoid adduction (AA) surgery not only medializes the paralyzed vocal fold, but increases the length of its membranous portion so that it more closely resembles the normal side. This could represent either real length change or out-of-plane rotation. Computed tomography scanning was performed on adult male cadaver larynges before and after the AA procedure to measure changes in length and spatial orientation of the vocal fold. Three dimensional coordinates of radiopaque markers on the anterior commissure, posterior glottic midline, and vocal processes were determined. The distance between the vocal processes was 3.9 mm before, and 0.8 mm after AA. The mean vocal fold length was 12.4 mm before, and 13.4 mm after AA (p = .14). The vocal process moved consistently caudally, an average of 3.5 mm (p = .02). The data suggest that clinically apparent vocal fold length changes with AA could be an illusion due to vertical displacement of the vocal process, and not actual lengthening. PMID- 9228854 TI - Endoscopic management of a combined laryngocele. AB - A 70-year-old women presented with hoarseness, foul sputum, and a softneck mass. Clinical and radiographic examination disclosed findings consistent with a combined internal and external laryngopyocele. Antibiotics, throat irrigations, and warm packs applied to the neck resulted in full resolution of the neck mass and subtotal regression of the supraglottic swelling. Endoscopic vestibulectomy was performed with a carbon dioxide laser; neither residual neck mass nor a tract leading to the extralaryngeal neck swelling was detected. The immediate and long term clinical course has been uneventful. This report is the first description of the definitive endoscopic management of a combined laryngocele. The pathogenesis of the laryngocele and the rationale and technique for endoscopic management are discussed. PMID- 9228855 TI - Epiglottic position after cricothyroidotomy: a comparison with tracheotomy. AB - Dysphagia is a known problem in patients with tracheotomy, but its association with cricothyroidotomy is not well studied. The purpose of this study was to evaluate dysphagia in patients with cricothyroidotomy and to determine if there is a reliable indicator of swallowing dysfunction in these patients. A review of charts for patients with modified barium swallow studies conducted at the New York University Medical Center Swallowing Disorders Center yielded three groups of patients: patients with cricothyroidotomy, patients with tracheotomy, and normal patients. There were 8 patients in each group. In all patients in the cricothyroidotomy group, there was a greater impairment of epiglottic displacement, laryngeal elevation, and upper esophageal opening than in the tracheotomy group. This problem with epiglottic displacement produced susceptibility to laryngeal penetration and, in turn, increased the risk of aspiration in those patients with cricothyroidotomy. After cricothyroidotomy tube removal, a return to normal epiglottic movement was observed within 2 months. One mechanism of swallowing dysfunction is impaired posterior displacement of the epiglottis over the glottic aperture. This impaired epiglottic motion appears to be related to restricted laryngeal elevation secondary to tethering of the larynx anteriorly at the site of the cricothyroidotomy. Additionally, we noted a decrease in the opening of the upper esophageal sphincter. PMID- 9228856 TI - Kaposi's sarcoma of the larynx. AB - Kaposi's sarcoma (KS) is a neoplastic vascular disorder, classically arising in the skin of the lower extremities. As a consequence of the acquired immunodeficiency syndrome (AIDS) epidemic, an increasing number of patients have been found to have KS. In AIDS patients, KS appears to exhibit a more diffuse nature and frequently affects the head and neck. Mucosal lesions are most often seen, commonly involving the oral cavity. Only rare cases of laryngeal involvement have been recorded in the literature. We report 2 cases of KS of the supraglottic larynx. Our first patient, an elderly man of Mediterranean descent, complained of voice change and throat discomfort. Endoscopy with biopsy for diagnosis allowed conservative treatment with chemotherapy. Our second patient was a younger man with AIDS who presented with symptoms of airway obstruction. Management with carbon dioxide laser epiglottectomy was successful in relieving that patient's symptoms. Although rare, KS may present in both healthy and immunocompromised patients, and must be considered in the differential diagnosis of all violaceous lesions of the larynx. PMID- 9228857 TI - Selective cochlear neurectomy for debilitating tinnitus. AB - Eighth nerve sections have been performed to control debilitating tinnitus, with various success rates (45% to 76%). Patients with a unilateral profound sensorineural hearing loss and disabling tinnitus perceived in that ear are candidates for such surgery. The concept of a selective cochlear neurectomy with preservation of the vestibular nerve is introduced with two case presentations. The indications for surgery, surgical technique, and results are described. Advantages of preserving the vestibular nerve fibers include the lack of postoperative vertigo and disequilibrium and thus a shorter length of hospital stay, and the conservation of a symmetric vestibular input, obviating the lengthy compensation process that might otherwise be needed, particularly in the elderly. A selective cochlear neurectomy for the control of debilitating tinnitus has proven to be successful in controlling tinnitus in the two patients presented, with the added advantage of preservation of their vestibular function. Further controlled studies are necessary to confirm the advantages and effectiveness of this technique. PMID- 9228858 TI - Interaction between oral alpha-streptococci and group A streptococci in patients with tonsillitis. AB - The incidence of oral alpha-streptococci with inhibitory activity against group A streptococci, as a defense mechanism against bacterial infection in the oral cavity, was investigated in 141 patients with streptococcal tonsillitis. The study population included both children (n = 79) and adults (n = 62). Infection by group A streptococci appeared to be more common in children than in adults, as the detection rates of inhibitory alpha-streptococci in healthy children (29.7%), as well as pediatric patients with tonsillitis (14.9%), were lower than those in adults (63.0%; p < .01). It is possible to consider oral alpha-streptococci with inhibitory activity to be among the indications for tonsillectomy in patients with streptococcal tonsillitis, since the detection rate of inhibitory alpha streptococci in surgical cases (10.9%) was significantly lower than that in nonsurgical cases (31.1%; p < .01). The high detection rate of these strains during the postoperative state supported the observation that the incidence of group A streptococcal infection was decreased postoperatively. Accordingly, it is useful to investigate bacterial interference between oral alpha-streptococci and group A streptococci in patients scheduled for tonsillectomy. PMID- 9228859 TI - Hearing loss (in nonoperated ears) in relation to age in osteogenesis imperfecta type I. AB - Hearing loss was studied in relation to age in nonoperated ears in a group of 142 subjects with autosomal dominant osteogenesis imperfecta type I, which was compared to that in a random subsample of 70 subjects. In the n = 142 group, particularly below the age of 30 years, considerable selection (ie, for ear surgery) had occurred on hearing loss. The hearing threshold increased gradually with age. A hearing loss of greater than 30 dB (Fletcher index) was observed for 51% of the subjects older than 20 years and younger than 60 years. The median hearing loss progressed from the 10th to the 45th years of life with an average annual threshold increase (ATI) of 1 dB/y (0.5 to 4 kHz) up to 1.7 dB/y (8 kHz). Sensorineural loss accounted for 0.6 dB/y ATI at 0.5 to 4 kHz and 1.3 dB/y ATI at 8 kHz; conductive loss accounted for 0.4 dB/y ATI at all frequencies. PMID- 9228860 TI - Narrowest (isthmus) portion of eustachian tube: a computer-aided three dimensional reconstruction and measurement study. AB - Nine normal human temporal bones from persons 16 to 88 years old were studied by computer aided three-dimensional reconstruction and measurement. The length of the eustachian tube (ET) lumen in three portions (from pharyngeal orifice to tympanic orifice: cartilaginous, junctional, and bony) averaged 23.6 +/- 4.3 mm, 3.0 +/- 1.9 mm, and 6.4 +/- 2.6 mm. The narrowest portion of the ET lumen was in the cartilaginous portion in all cases: 20.5 +/- 4.2 mm from the pharyngeal orifice and 3.1 +/- 1.6 mm from the pharyngeal margin of the junctional portion. The cross-sectional area of the narrowest portion was 0.65 +/- 0.2 mm2. The tendon of the tensor veli palatini muscle (TVPM) inserted into the lateral lamina in the narrowest portion of the ET lumen in five of nine cases. These results suggest that contraction of the TVPM opens the narrowest portion of the ET lumen to ventilate the middle ear and that this portion also plays a role in protecting the middle ear. PMID- 9228861 TI - Recurrent pleomorphic adenoma of the parotid gland involving the osseous external auditory canal. With a note on the foramen of Huschke. AB - A recurrence of a parotid pleomorphic adenoma presenting as an external ear canal mass is reported. The route of extension was radiographically documented as an incompletely closed foramen of Huschke. The historical and developmental features of this foramen and the mechanisms of tumor extension as they relate to it are discussed. PMID- 9228862 TI - Cross-innervation of the thyroarytenoid muscle by a branch from the external division of the superior laryngeal nerve. AB - The neuroanatomy of the larynx was explored in seven dogs to assess whether there is motor innervation to the thyroarytenoid (TA) muscle from the external division of the superior laryngeal nerve (ExSLN). In 3 animals, such innervation was identified. Electrical stimulation of microelectrodes applied to the ExSLN resulted in contraction of the TA muscle, indicating that this nerve is motor in function. This was confirmed by electromyographic recordings from the TA muscle. Videolaryngostroboscopy revealed improvement in vocal fold vibration following stimulation of the ExSLN compared to without it. Previously, the TA muscle was thought to be innervated solely by the recurrent laryngeal nerve. This additional pathway from the ExSLN to the TA muscle may have important clinical implications in the treatment of neurologic laryngeal disorders such as adductor spasmodic dysphonia. PMID- 9228863 TI - Ototoxic effect of erythromycin on cochlear potentials in the guinea pig. AB - The mechanism of hearing loss due to the administration of intravenous erythromycin was investigated in the albino guinea pig, and it was found for the first time that this drug causes cochlear dysfunction. The endocochlear potential (EP) and the cochlear microphonics (CM) recorded at the first cochlear turn transiently decreased when erythromycin was administered intravenously at dosages of 100 and 150 mg/kg. The averaged maximum decrease in EP was 16 mV (n = 5) and 33 mV (n = 5) for 100 and 150 mg/kg, respectively. The maximum decrease in the CM was about 25% when the EP reached its lowest value with the injection of 150 mg/kg. A complete recovery of the EP and CM ensured within 20 minutes after each erythromycin dose. The perilymphatic perfusion of 3 mmol/L of erythromycin decreased the EP and CM; however, in contrast to the intravenous administration, the decrease of the CM was nearly complete and both the EP and CM were irreversible. Hearing loss due to intravenously administered erythromycin could likely be attributle to the transient dysfunction of the stria vascularis, although concomitant dysfunction of the central auditory pathway cannot be excluded. PMID- 9228864 TI - Effects of platelet activating factor on vascular permeability of the middle ear mucosa. AB - Platelet activating factor (PAF), a potent inflammatory mediator, seems to play a significant role in the pathogenesis of otitis media with effusion (OME), along with other inflammatory mediators such as leukotrienes and prostaglandins. The purpose of this study was to investigate the effect of PAF on the vascular permeability of middle ear mucosa, in an experimental OME model using chinchillas. We injected PAF in doses of 1, 4, 8, and 16 micrograms and normal saline as a control into the bullae of chinchillas. Vascular permeability was measured by the Evans blue vital dye technique. All the PAF-injected animals showed a significant increase in middle ear vascular permeability compared to the control group. This study demonstrated that PAF in the middle ear cavity contributes significantly to the development of OME by increasing the vascular permeability of the middle ear mucosa. PMID- 9228865 TI - Imaging case study of the month. Congenital cholesteatoma isolated to the mastoid. PMID- 9228866 TI - Human immunodeficiency virus-1 infection of the lymphoid tissues of Waldeyer's ring. AB - Human immunodeficiency virus-1 (HIV-1) infection is a fatal retroviral infection that may first present clinically as enlargement of the lymphoid tissues of Waldeyer's ring. These tissues are a major site of viral replication. The presence of the virus in these tissues causes a unique constellation of diagnostic histopathologic features, including florid follicular hyperplasia, follicle lysis, and productively HIV-1-infected multinucleated giant cells of probable dendritic cell origin. Serologic evaluation is confirmatory of HIV infection. With the recent advances in antiretroviral chemotherapy, the early institution of which may significantly prolong life and disease-free interval, the recognition of the clinical and pathologic parameters of HIV-related enlargement of Waldeyer's ring tissues is essential. PMID- 9228867 TI - Extraesophageal pediatric reflux: 24-hour double-probe pH monitoring of 222 children. AB - Although extraesophageal gastric reflux has been implicated as a cause of many pediatric airway and respiratory diseases, its prevalence in these conditions remains unknown due to the relative lack of sensitivity and/or specificity of traditional reflux testing methods. A prospective study of 222 children (ages 1 day to 16 years) was performed with 24-hour double-probe (simultaneous esophageal and pharyngeal) pH monitoring. Seventy-six percent (168/222) of the study population had abnormal findings in either one or both of the pH probes. Of those, 46% (78/168) had pharyngeal reflux (extraesophageal gastric acid documented by the pharyngeal probe), despite having normal esophageal acid exposure times according to the esophageal probe. Thus, had the pharyngeal probe not been used, 46% of the children with documented extraesophageal (pharyngeal) reflux would have been falsely presumed to have normal reflux parameters. Patients with laryngeal abnormalities, pulmonary abnormalities, and emesis had significantly more pharyngeal acid reflux (p < .001) than patients with nonrespiratory symptoms. These data suggest that extraesophageal reflux may be underestimated by single-probe intraesophageal monitoring alone, and that laryngopharyngeal reflux may play a role in the pathogenesis of the conditions studied. PMID- 9228868 TI - Strength, endurance, and work capacity after muscle strengthening exercise in postpolio subjects. AB - OBJECTIVE: To determine whether a 12-week home quadriceps muscle strengthening exercise program would increase muscle strength, isometric endurance, and tension time index (TTI) in postpolio syndrome subjects without adversely affecting the surviving motor units or the muscle. DESIGN: A longitudinal study to investigate the effect of a 12-week exercise program on neuromuscular function and electromyographic variables. SETTING: Neuromuscular laboratory of a university hospital. SUBJECTS: Seven subjects were recruited from a cohort of 12 subjects who had participated in a previous exercise study. All subjects had greater than antigravity strength of the quadriceps. Upon completion of a postpolio questionnaire, all acknowledged common postpolio syndrome symptoms such as new fatigue, pain, and weakness; 6 of the 7 acknowledged new strength decline. INTERVENTION: On Mondays and Thursdays subjects performed three sets of four maximal isometric contractions of the quadriceps held for 5 seconds each. On Tuesdays and Fridays subjects performed three sets of 12 dynamic knee extension exercises with ankle weights. MAIN OUTCOME MEASURES: Neuromuscular variables of the quadriceps muscles were measured at the beginning and completion of the exercise program and included: isokinetic peak torque (ISOKPT, at 60 degrees/sec angular velocity) and total work performed of four contractions (ISOKTW), isometric peak torque (MVC), endurance (EDUR, time subject could hold isometric contraction at 40% of the initial MVC), isometric tension time index (TTI, product of endurance time and torque at 40% of MVC), and initial and final ankle weight (WGT, kg) lifted. Electromyographic variables included: fiber density (FD), jitter (MCD), and blocking (BLK) from single fiber assessment and median macro amplitude (MACRO). Serum creatine kinase (CK) was also measured initially and at 4-week intervals throughout the study. RESULTS: The following variables significantly (p < .05) increased: WGT by 47%, ISOKPT, 15%, ISOKTW, 15%; MVC, 36%; EDUR, 21%; TTI, 18%. The following variables did not significantly (p > .05) change: FD, MCD, BLK, MACRO, and CK. CONCLUSIONS: This home exercise program significantly increased strength, endurance, and TTI without apparently adversely affecting the motor units or the muscle, as the EMG and CK variables did not change. PMID- 9228869 TI - Prostaglandin E2 measurements: their value in the early diagnosis of heterotopic ossification in spinal cord injury patients. AB - OBJECTIVE: To look for a possible relation between the occurrence of heterotopic ossification (HO) and the modifications of the 24-hour prostaglandin E2 (PGE2) urinary excretion. DESIGN: A 5-year prospective study to determine the 24-hour urinary excretion of PGE2 by radioimmunoassay with specific antisera not cross reacting with TXA2, TXB2, 15-keto-PGE2 alpha, PGI2, 6-keto-PGF1 alpha. SETTING: The laboratory of a division of endocrinology and diabetology of a university hospital. PATIENTS: Of 262 acute spinal cord injury patients screened, 44 were eligible for the study. INTERVENTIONS: Serial diagnostic quantitative bone scannings with technetium 99m Tc methylene diphosphate (99mTc-MDP) and therapeutic assessment of radiotherapy and indomethacin. MEAN OUTCOME MEASURE: Hypothetical increase of PGE2 before and during HO formation. RESULTS: Of 44 patients, 8 developed an HO (18.8%) with concomitant marked increase of the PGE2 excretion for as long as the HO had not reached maturity. The results of the radiotherapy were inconclusive. Indomethacin was shown to be efficacious in holding back or slowing down the HO evolution. CONCLUSIONS: Measurement of the 24 hour PGE2 urinary excretion appears to be a valuable indicator in the early diagnosis of HO. Indomethacin should be considered as an alternative to other existing therapies. PMID- 9228870 TI - Parathyroid hormone suppression in spinal cord injury patients is associated with the degree of neurologic impairment and not the level of injury. AB - OBJECTIVE: To demonstrate that after spinal cord injury (SCI) suppression of the parathyroid-vitamin D axis is associated with the degree of neurologic impairment and not the level of injury. DESIGN: A retrospective analysis of clinical and biochemical data obtained from hospital records of patients with SCI compared to a control group of patients with traumatic brain injury (TBI). SETTING: The inpatient rehabilitation unit of a tertiary care hospital. SUBJECTS: The medical records of 82 consecutive admissions to the rehabilitation unit with a diagnosis of SCI or TBI were reviewed. Patients with SCI were classified by the American Spinal Injury Association (ASIA) impairment scale and then grouped based on the completeness and level of injury. MAIN OUTCOME MEASURE: Comparisons of serum parathyroid hormone (PTH), 25-hydroxyvitamin D, and 1,25-dihydroxyvitamin D (1,25 D) were planned. Multiple comparisons were performed for total and ionized serum calcium levels, serum phosphorus levels, and 24-hour urinary calcium excretion rates to reflect changes in mineral homeostasis. Multiple comparisons were also performed for serum albumin, prolactin, thyroid function tests, and AM cortisol levels, as well as 24-hour urinary urea nitrogen and cortisol excretion rates to reflect metabolic responses to stress. RESULTS: Patients with SCI had significant suppression in PTH (p < .000009) and 1,25-D (p < .02) levels with elevated phosphorus (p < 0.03) and prolactin (p < .03) levels compared to patients with TBI. Also, more patients with SCI were hypoalbuminemic (p < .003) than patients with TBI. Patients with complete SCI (ASIA A) had more suppressed PTH (p < .03) and higher urinary urea nitrogen (p < .05) levels than SCI patients with incomplete injuries (ASIA B-D). Patients with complete, but not incomplete, SCI had lower albumin levels than patients with TBI (p < .05). These differences were not found between patients with tetraplegic and paraplegic SCI. ASIA motor scores did not correlate with any of the measured parameters but when used as a covariate did abolish differences in PTH and 1,25-D among the study groups by ANOVA. CONCLUSION: In patients with SCI, the degree of neurologic impairment, and not the level of injury, is associated with PTH suppression and markers of metabolic stress. PMID- 9228871 TI - Coronary heart disease risk indicators, aerobic power, and physical activity in men with spinal cord injuries. AB - OBJECTIVE: To compare the lipid and (apo-)lipoprotein profile and blood pressure of men with long-standing spinal cord injuries (SCI) to those of an age-matched able-bodied (AB) population, and to assess the most important determinants of this profile and blood pressure. DESIGN: A cross-sectional study of persons with chronic SCI residing in the community. SETTING: Tests were performed in a university research laboratory. SUBJECTS: Thirty-seven men (age 37.4 +/- 12.0 yrs) with longstanding (14.7 +/- 8.6 yrs) SCI ranging from level C4/5 to L5 volunteered to participate. Comparisons were made with published data from 3,498 AB men, age 20 to 59 yrs, from the same country. MAIN OUTCOME MEASURES: Lipid and lipoprotein profile (total cholesterol [TC], high-, low-, and very low-density lipoprotein cholesterol [HDL-C, LDL-C and VLDL-C, respectively], and triglycerides [TG]), as well as aerobic power, activity level, anthropometric variables, and blood pressure. Multiple regression analyses assessed the most important determinants of the lipid and blood pressure profile. RESULTS: None of the lipid variables were related to the lesion level. TC, HDL-C, and TC/HDL-C were not significantly different from the AB population. The most important determinants of TC, LDL-C, and the ratios TC/HDL and HDL-C/LDL-C were age, smoking behavior, and activity level. Aerobic power was not an important determinant of any lipid or (apo-)lipoprotein or blood pressure. CONCLUSION: Men with long-standing SCI do not appear to have an essentially different coronary heart disease risk profile compared with AB persons. Modifiable risk factors such as activity level, smoking, alcohol consumption, body mass index, and adipose tissue were more important than lesion level and aerobic power in the determination of the lipid and lipoprotein profile, suggesting several potential interventions. PMID- 9228872 TI - Functional outcomes attained by T9-12 paraplegic patients with the walkabout and the isocentric reciprocal gait orthoses. AB - OBJECTIVE: To compare the functional outcomes attained by persons with paraplegia using the Walkabout Orthosis (WO) and the Isocentric Reciprocal Gait Orthosis (IRGO). DESIGN: A randomized crossover design. PATIENTS: Ten subjects with complete lesions between T9-T12. INTERVENTIONS: Over two 8-week periods, subjects were taught to use each orthosis in conjunction with elbow crutches. MAIN OUTCOME MEASURES: After each 8-week training period, subjects were assessed on their ability to perform five different sets of key skills associated with functional ambulation. RESULTS: There were no differences between orthoses in the ability of subjects to don and doff the orthoses, get up and down stairs and curbs, or walk on a flat surface. Subjects required significantly more assistance when using the WO to walk over inclined surfaces (median IRGO = "independent," median WO = "minimal assistance"; p = .03) but less assistance when using the WO to get from sitting to standing and standing to sitting (median IRGO = "moderate assistance," median WO = "minimal assistance"; p = .03). In addition, subjects walked significantly faster with the IRGO both on the flat (mean IRGO = .34 m/sec +/- .18, mean WO = .14 m/sec +/- .12; p = .002) and on inclined surfaces. CONCLUSIONS: Although it is easier to stand up and sit down with the WO, the IRGO facilitated a faster and more independent gait. Neither orthosis enabled subjects to be fully independent in the key skills necessary for functional ambulation after 8 weeks of training. PMID- 9228873 TI - Physiologic responses during functional electrical stimulation leg cycling and hybrid exercise in spinal cord injured subjects. AB - OBJECTIVES: (1) To determine if a hybrid exercise (leg plus arm) training program performed immediately after functional electrical stimulation (FES) leg cycle exercise (LCE) training would further improve aerobic capacity when compared with FES leg cycle training alone, and (2) to compare the submaximal responses occurring during both FES-LCE alone and hybrid exercise in the same SCI subjects. DESIGN: Nonrandomized control trial whereby subjects act as their own control. SETTING: Outpatient rehabilitation in a primary care hospital. PATIENTS: A volunteer sample (n = 11) of men 20 to 50 years old with complete spinal cord injury, free from cardiovascular and metabolic disease with spasticity. INTERVENTIONS: Three phases of exercise training: phase I, progressive FES-LCE to 30 minutes of exercise (n = 11); phase II, 35.2 +/- 16.2 sessions of FES-LCE (n = 11); phase III, 41.4 +/- 17.7 30-minute sessions of hybrid exercise (n = 8). MAIN OUTCOME MEASURES: (1) Aerobic capacity-a further increase after hybrid exercise when compared with FES-LCE alone; (2) submaximal physiologic parameters (oxygen uptake [VO2], heart rate [HR], blood lactate [BLa-])-measurement of these during constant work rate exercise and a training effect. RESULTS: VO2 (the body's ability to utilize oxygen) significantly improved (p < .05) after both FES-LCE and then further after hybrid training. Hybrid exercise training resulted in significantly (p < .05) greater work rates and VO2 values than both FES-LCE at baseline and training work rates. CONCLUSION: These subjects demonstrated that hybrid exercise performed twice a week provided sufficient intensity to improve aerobic capacity and provide a medium whereby patients with SCI can burn more calories than via FES-LCE alone. This has important implications for improving the health and fitness levels of individuals with SCI and may ultimately reduce their risk of cardiovascular disease. PMID- 9228874 TI - Asymmetry of gait initiation in hemiparetic stroke subjects. AB - OBJECTIVE: To investigate symmetry of gait initiation in healthy and hemiparetic subjects. DESIGN: Survey. SETTING: Kinematic laboratory affiliated with a hospital-based department of rehabilitation. PATIENTS OR OTHER PARTICIPANTS: Ten healthy and 14 hemiparetic stroke subjects starting five times with their right and left leg, respectively. MAIN OUTCOME MEASURES: Duration of defined periods, step length, center of pressure, and center of mass were recorded and calculated using two triaxial force plates, contact switches, and a video camera system. RESULTS: Healthy subjects displayed a high degree of independence of kinetic and kinematic parameters of the starting limb. Hemiparetic patients showed differences with respect to the starting limb: when starting with the nonaffected leg, the swing period and step length was shorter and the center of pressure displayed a more marked medio-lateral sway with no corresponding initial movement of the center of mass; when starting with the affected leg the movement pattern of the center of pressure and center of mass was comparable to that of normal subjects. CONCLUSIONS: The trajectories of the center of pressure and center of mass and the symmetry parameters are in accordance with a higher degree of uncertainty when starting with the non-affected limb in hemiparetic subjects. PMID- 9228875 TI - Systematic and random error in repeated measurements of temporal and distance parameters of gait after stroke. AB - OBJECTIVE: To obtain intersession estimates of error for temporal and distance (TD) parameters of gait in a sample of stroke patients undertaking inpatient rehabilitation. DESIGN: Thirty-one stroke patients were measured with an instrumented footswitch system (after a median of 46 days poststroke; interquartile range = 26 to 63) walking over a 10-meter distance a total of four times on 3 consecutive days. Two familiarization walks provided intrasession retest data. RESULTS: Metric estimates of systematic and random error have been provided for obtained TD parameters. Proportional indices of reliability (ICC [2,1] and Pearson's r) were generally high, ranging from .72 to .94. CONCLUSION: By quantifying systematic and random error associated with the process of repeated measurements, criteria have been provided for evaluating change in TD variables during rehabilitation. Although error for gait velocity was small relative to individual differences in the stroke group, it was large relative to levels of change derived from measurements reported during typical periods of rehabilitation. Serial measurements of gait during rehabilitation may be better than two consecutive measurements. This study highlights the need to interpret estimates of error according to the purpose of measurement. PMID- 9228877 TI - Spectral characteristics of postural control in elderly individuals. AB - OBJECTIVE: To examine the feasibility of using spectral analysis techniques to identify potential biomarkers of diminished postural control in elderly individuals. DESIGN: Data from spectral signatures (derived from postural sway) of 21 young adults and 42 elderly individuals classified as "high" or "low" risk with regard to functional balance capacity were analyzed using Risk Category (3) x Sensory Condition (3) multivariate analyses of variance. Postural control was challenged by varying the visual conditions under which individuals stood on a measurement platform. RESULTS: Results indicated that measures of central tendency and dispersion of the spectral frequency distribution from medial lateral components of sway (but not antero-posterior sway) clearly differentiated between "high" and "low" risk elderly. Low risk elderly were not different from young adults. High risk elderly exhibited greater dispersion and lower mean frequency than other groups. CONCLUSIONS: Differences in spectral characteristics of medial-lateral components of sway were more related to risk category than to age. Elderly persons with high functional balance capacity displayed characteristics similar to those of young adults. Thus, spectral frequency analysis techniques may be a clinically useful tool for identifying individuals potentially at risk of falling. PMID- 9228876 TI - Hyperventilation effect on postural sway. AB - OBJECTIVE: To examine the effect of voluntary hyperventilation (HV) on postural sway. DESIGN: Crossover controlled, experimental study. SETTING: Human movement and balance clinical research unit. SUBJECTS: Four different groups of normal subjects (n = 6, 6, 7, and 9) and patients with bilateral absence of vestibular function (n = 9). INTERVENTION: Partial carbon dioxide pressure (tc-PCO2) was measured transcutaneously with surface electrodes. Body sway was measured with a force platform immediately after maximal voluntary HV for 30 to 90 seconds. Recordings were obtained with eyes open and eyes closed, standing on the platform and on foam-rubber, and after head or body movements. MAIN OUTCOME MEASURE: Postural sway. RESULTS: HV increased body sway in all conditions, but the effects were more intense when subjects were standing directly on the platform surface with their eyes closed. Recordings after HV of 30, 60, and 90 sec in normal subjects showed that although CO2 levels were inversely related to the duration of HV, body sway did not increase further. HV also increased sway after active movements by the subjects. The main sway increase was in sway area and mean and maximal deviations but less for mean sway velocity. HV preferentially increased low-frequency sway oscillations. These effects were also present in labyrinthine defective subjects. CONCLUSIONS: HV increases body sway, but the relationship between CO2 levels and degree of unsteadiness is not linear. The dizziness reported by patients with HV syndrome may be partly caused by objective unsteadiness. The presence of HV-induced unsteadiness in patients with absent vestibular function indicates that the effects of HV are not mediated by the labyrinth. PMID- 9228878 TI - Generation II knee bracing for severe medial compartment osteoarthritis of the knee. AB - OBJECTIVE: To investigate the clinical efficacy of the Generation II (G II) knee brace, a newly developed knee orthosis, on patients experiencing severe medial compartment osteoarthritis (OA) of the knee. DESIGN: Case series. SETTING: A national medical and pharmaceutical hospital in Japan. PATIENTS: Twenty primary OA subjects (excluding those with secondary OA), all older than 55 years of age and experiencing only knee joint problems, were selected according to their ability to walk more than 500 meters independent of support. These patients had arthritis in both knees and no less than one half of normal joint space remaining as revealed by roentgenogram studies. The more severely affected side was selected for bracing. INTERVENTIONS: For 12 months, each patient wore a G II knee brace on the affected knee on a daily basis, removing it only at night. To evaluate the effects of G II OA brace alone, additional use of new oral drugs or any other treatment was prohibited from 1 month before application of the G II OA brace and throughout the trial period. MAIN OUTCOME MEASURES: Clinical efficacy was evaluated using the Japan Orthopaedic Association's knee scoring system. X ray evaluation was performed with patients standing on one leg. A dynamometer was used to evaluate isokinetic quadriceps muscle strength. The center of gravity was measured using an X-Y recording. Clinical evaluation was performed every 2 months thereafter. Final evaluation was at 12 months. RESULTS: Nineteen of the 20 patients answered that they experienced significant pain relief. Knee pain scores on walking increased from 18.0 to 21.5 and on ascending and descending stairs increased from 12.8 to 15.8. The femorotibial angle decreased in 12 of the patients, and the mean angle decreased from 185.1 degrees before application to 183.7 degrees with the brace on at the final observation period. In addition, isokinetic quadriceps muscle strength increased from an average of 36.8 Nm to 42.8 Nm for all patients. In 17 patients, quadriceps muscle strength increased, while it decreased in 2 and remained the same in 1. Finally, lateral movement of the center of gravity decreased compared with before G II application in all patients. CONCLUSION: G II bracing is a beneficial treatment for severe medial OA of the knee. PMID- 9228879 TI - Prosthesis satisfaction outcome measurement in pediatric limb deficiency. AB - OBJECTIVE: To describe the development and initial psychometric properties of a new outcome measure to assess satisfaction with prosthesis in children with limb deficiencies. DESIGN: Parents of children with limb deficiency were surveyed during routine outpatient clinic visits. SETTING: Two outpatient pediatric clinics. PARTICIPANTS: Ninety-seven parents of children with limb deficiency aged 1 to 17 years. MAIN OUTCOME MEASURE: The newly developed Child Amputee Prosthetics Project-Prosthesis Satisfaction Inventory (CAPP-PSI). RESULTS: Internal consistency reliability is high. Zero-order correlations with prosthesis wear/use patterns and with parent ratings of prosthesis appearance provide support for the construct validity of the CAPP-PSI. CONCLUSION: The CAPP-PSI is a promising, brief, parent-administered inventory for the assessment of prosthesis satisfaction in children with limb deficiency. It may be useful in future research for predicting prosthesis wear and use patterns in this population. PMID- 9228880 TI - Home alone: the role of cognition in return to independent living. AB - OBJECTIVE: To identify unique predictors of the ability to return to living alone in geriatric patients undergoing medical rehabilitation. DESIGN: Of 900 consecutive geriatric patients entering medical rehabilitation, 372 were identified as living alone before admission. Data were collected on functional status, cognition, demographics, and discharge disposition. SETTING: A freestanding medical rehabilitation facility. All patients were admitted to a geriatric rehabilitation unit. PATIENTS: Patients aged 60 to 99, identified as having lived alone before admission, were included. As standard procedure, patients underwent functional and cognitive assessment, and medical records were reviewed. MAIN OUTCOME MEASURE: Logistic regression analysis was used to evaluate predictors of discharge disposition, including demographic variables, medical burden, the Functional Independence Measure (FIM), and the Dementia Rating Scale (DRS). RESULTS: Both the FIM and DRS provided significant and unique variance in prediction of discharge disposition. Patients discharged home alone performed similarly to those discharged with supervision on FIM motor items but higher on FIM social cognition items, emphasizing the strong role of cognition in discharge disposition. Patients discharged home alone scored above suggested cutoff scores on the DRS, indicating generally intact cognitive functioning, whereas those discharged with supervision scored below suggested cutoffs. CONCLUSION: Results emphasize the importance of cognition in the ability to return to completely independent living after medical rehabilitation in geriatric patients. PMID- 9228881 TI - Computer-aided video analysis of vertebrofemoral motion during toe touching in healthy subjects. AB - OBJECTIVE: Despite widespread use of the toe touch test, the relative contribution from vertebral and hip movements has not been clearly established, largely because of unsatisfactory measurement techniques. This study aimed to reinvestigate the kinematics of toe touching by combining computerized videotape analysis with a new model of reference marker placement. METHOD: Twenty-two subjects were videotaped during active toe touching from upright standing. Computer software was then used to derive the sagittal thoracic, lumbar, and hip angles at .02-sec intervals throughout the movement. RESULTS: Hip flexion was directly proportional to toe touch distance (TTD) (r2 = .71) but not lumbar flexion (r2 = .17) or thoracic (r2 = .20) excursion. On average there was .8 degree of thoracolumbar flexion for every 1 degree of hip flexion; however, there were wide variations between subjects. In 19 of 22 subjects the thoracic spine flexed and extended relatively equal amounts during the test resulting in a small total thoracic excursion of 4.8 degrees flexion in unsuccessful toe touchers and 4.0 degrees extension in successful toe touchers. CONCLUSION: The separate contributions of hip, lumbar, and thoracic mobility to toe touching or any other vertebrofemoral motion can only be accurately determined by a measurement strategy that uses the plane of the pelvis to separate vertebral from hip motion and uses tangents at the limits of the thoracic and lumbar regions to separate lumbar from thoracic motion. Using this model the authors found that TTD is not a reliable indicator of either vertebral or hip mobility. PMID- 9228882 TI - Systemic sclerosis (scleroderma): an integrated challenge in rehabilitation. AB - Systemic sclerosis (SSc), a multisystem disease involving the microvascular system and the connective tissue, is considered one of the most difficult rheumatic diseases to treat. The natural history of the disease evolves from an edematous to a scleroatrophic phase following two different temporal patterns: acute or chronic. The former leads to early death, and the latter evolves slowly toward severe disability that deserves rehabilitative intervention. Despite the poor prognosis, recent improvements in diagnosis and treatment have led to longer patient survival, thus increasing the need to intervene against the development of tissue fibrosis and contractures by using appropriate integrated rehabilitation programs. This article does not review the medico-pharmacological management of visceral manifestations of the disease. Rather, it is divided into six parts, which include analyses of the changes in skin, joints and tendons, and muscle induced by SSc; examination of the existing literature on rehabilitation strategies and treatments; discussions of the pain and peripheral sensory-motor system involvement that are present to a greater or lesser extent in almost all patients and influence not only the duration and outcome of rehabilitation but also the patient's family, social life, and working ability; and consideration of ergonomic and occupational interventions. No controlled studies have been done on the few rehabilitation guidelines and specific protocols identified, so it must be emphasized that this article is a summary of opinions expressed in the literature and the authors' own findings. Particularly lacking are studies on such aspects as ergonomics, work intervention, or the management of sexual dysfunction. Experience gained in the rehabilitation of skin burns and other rheumatic diseases forms the basis for a logical approach to SSc patients. PMID- 9228883 TI - Heterotopic ossification as a complication of toxic epidermal necrolysis. AB - The development of heterotopic ossification (HO) as a complication of toxic epidermal necrolysis (TEN) has not been previously reported. TEN, also known as Lyell's syndrome, is a rare but serious skin disorder that typically occurs after the administration of drugs, especially sulfonamides, barbiturates, phenytoin, and nonsteroidal anti-inflammatory agents. TEN is characterized by the development of large fluid-filled bullae with separation of large sheets of skin. Complications of TEN can include extensive denudation of skin with dehydration and electrolyte abnormalities, gastrointestinal hemorrhage, acute tubular necrosis, secondary infection of denuded skin, pneumonia, bacterial conjunctivitis, keratitis, and septic infarcts of internal organs. We report a case of HO in a patient with TEN after treatment with trimethoprim sulfamethoxazole. A 49-year-old man developed an erythematous rash, bullae, fever, and extensive skin loss consistent with a diagnosis of TEN. He was intubated for complications of TEN (pneumonia) and maintained on bed rest for several weeks. In addition, he developed HO that resulted in multiple joint contractures. He was treated with aggressive range of motion by physical therapy, surgical resection of the HO followed by radiation to both elbows, right hip, and right knee. Postoperative outpatient rehabilitation enabled improved function in his mobility and activities of daily living. HO is known to occur after spinal cord and brain injuries and burns. It has not been reported to occur after TEN. Our experience with this case suggests that HO may merit inclusion into the list of complications of TEN. PMID- 9228884 TI - Autonomic dysfunction as the presenting feature of Guillain-Barre syndrome. AB - Autonomic dysfunction has been demonstrated in various conditions associated with peripheral neuropathy such as acute intermittent porphyria, amyloidosis, and Guillain-Barre syndrome (GBS). In the latter, hypertension is an associated complication that typically occurs after neurological signs are already present. We report a case of a patient with autonomic dysfunction as the presenting feature who was admitted to the coronary unit with chest pain and hypertension. Subsequently, he developed progressive symmetric muscle, weakness, sensory changes, and areflexia. GBS was then diagnosed based on the clinical picture, albuminocytologic dissociation in the cerebrospinal fluid, and electrodiagnostic abnormalities suggestive of demyelinative polyneuropathy with conduction block. Few cases in the literature have reported autonomic dysfunction as the presenting feature of GBS, such as in this case. In a previously asymptomatic patient, acute onset of autonomic dysfunction should alert the physician to the possibility of an acute polyneuropathy, such as GBS. PMID- 9228885 TI - Total knee arthroplasty in a patient with bilateral Charcot knees. AB - Neuropathic arthropathy (Charcot joint) is a progressive and degenerative process resulting from underlying neurovascular and neurotraumatic deficits. Diabetes mellitus is now the most common cause of Charcot joint. A marked predilection for the tarsometatarsal, tarsal, and ankle joints occurs. Involvement of large weight bearing joints such as the knee is rare. When the knee is involved, and conservative treatment fails, standard surgical intervention often involves arthrodesis. Arthroplasty is relatively contraindicated. The authors report a case of a 61-year-old, diabetic-woman with bilateral Charcot knees who successfully completed a rehabilitation program and achieved independence after left knee arthrodesis and right total knee arthroplasty. PMID- 9228886 TI - Isolated traumatic denervation of the extensor pollicis longus. AB - This report describes a case of isolated denervation of the extensor pollicis longus (EPL), which has not previously been reported. The patient was a healthy 13-year-old boy who sustained multiple lacerations to the left forearm secondary to a dog bite. No motor responses were evident in the EPL, but the remainder of the exam findings were normal. Electromyography confirmed complete isolated denervation of the EPL. This patient was managed nonoperatively and had no significant functional deficits at final follow-up. PMID- 9228887 TI - Caffeine and chronic back pain. PMID- 9228888 TI - Excitotoxicity and neuroprotection. PMID- 9228889 TI - Sleep disturbances and suicidal behavior in patients with major depression. AB - BACKGROUND: The purpose of this study was to examine the association between sleep disturbances and suicidal behavior in patients with major depression (N = 113). METHOD: The sleep symptomatology of each patient was ascertained from the Schedule for Affective Disorders and Schizophrenia (SADS) questions concerning sleep in the section on major depression. The patients were retrospectively classified as having hypersomnia (N = 20), insomnia (N = 69), and no sleep disturbance (N = 24). The SADS suicide subscale was used to rate the severity of active suicidality. RESULTS: The patients with hypersomnia and insomnia had significantly (p < .05) higher scores on the SADS suicide subscale than those without sleep disturbance. We also found that the patients with insomnia and hypersomnia were significantly (p < .001) more likely to become suicidal than the others. CONCLUSION: These data demonstrate that both insomnia and hypersomnia are associated with suicidal behavior in patients with major depression. PMID- 9228891 TI - Hypouricemia in chronic schizophrenic patients with polydipsia and hyponatremia. AB - BACKGROUND: Polydipsia is a common disorder among chronic psychiatric patients. Impaired water excretion due to enhanced action and secretion of antidiuretic hormone has been reported in hyponatremic patients with polydipsia. Hypouricemia coexisting with hyponatremia is a hallmark of the syndrome of inappropriate secretion of antidiuretic hormone (SIADH). The transitory coexistence of hyponatremia and hypouricemia in patients with polydipsia-hyponatremia syndrome is presented. METHOD: We examined the course of hypouricemia and hyponatremia in three schizophrenic patients with a long-standing history of polydipsia resulting in the presence of these conditions. In addition, we investigated the renal clearance of uric acid in five polydipsic patients without a previous history of water intoxication or hyponatremia (simple polydipsia). RESULTS: Both hyponatremia and hypouricemia were demonstrated in the presence of SIADH in one patient, during an episode of acute water intoxication in another, and in association with chronic hyponatremia in a patient who was following the target weight procedure. Elevated fractional excretion of uric acid percentage (FEUA%) was detected in two patients. These states appeared to be episodic or transitory. In the five patients with simple polydipsia, serum uric acid concentrations and FEUA% were maintained within the normal range. CONCLUSION: Altered uric acid regulation that resembles SIADH is present in patients with polydipsia hyponatremia syndrome. Monitoring the uric acid concentration and FEUA% in polydipsic patients may be useful in identifying those patients with transiently impaired water excretion. PMID- 9228890 TI - Clozapine withdrawal resulting in delirium with psychosis: a report of three cases. AB - BACKGROUND: Withdrawal symptoms for typical antipsychotics are generally mild, self-limited and do not include development of psychotic symptoms. In contrast, withdrawal symptoms for clozapine can be severe with rapid onset of agitation, abnormal movements, and psychotic symptoms. Different pathophysiologic etiologies have been suggested for these severe symptoms, including dopaminergic supersensitivity and rebound. METHOD: Three case reports of clozapine withdrawal symptoms are presented. A review of previous case reports and discussion of the etiology of withdrawal symptoms of typical antipsychotics and clozapine are provided. RESULTS: These three patients developed delirium with psychotic symptoms that resolved rapidly and completely upon resumption of low doses of clozapine. CONCLUSION: The severe agitation and psychotic symptoms after clozapine withdrawal in these three patients were due to delirium, perhaps the result of central cholinergic rebound. The withdrawal symptoms and delirium resolved rapidly with resumption of low doses of clozapine. Severe withdrawal symptoms can probably be avoided by slowly tapering clozapine and/or simultaneously substituting another psychotropic with high anticholinergic activity, such as thioridazine. PMID- 9228892 TI - Depression after stroke: an investigation through catamnesis. AB - BACKGROUND: In the early stage of stroke, depression appears to be linked to certain brain areas. The study evaluated the importance of the side of the lesion in depressed patients 3 years after their first stroke. METHOD: Patients who had suffered a stroke and been discharged after rehabilitation were identified by hospital records. We interviewed 180 patients at home. Demographic as well as socioeconomic data were collected. The North-western University Disability Scale, the Beck Depression Inventory (BDI), the Relatives' Stress Scale, and the Social Dysfunction Rating Scale were applied. The diagnosis was confirmed for each patient by a clinical assessment according to the ICD-10 criteria. Patients with previous psychiatric treatment, comprehension problems, or severe hemi inattention were excluded. RESULTS: By using a score of 14 on the BDI as a cutoff, 62 patients (34%) proved to be affected by depressive disorders. Clinical records showed that the location of the lesion was in the right hemisphere for 37 patients and in the left hemisphere for 25 patients. Statistical analysis of the mean scores obtained in this subgroup of depressed patients showed (1) no significant relation between depression and the hemispheric location of the lesion or between depression and level of education; (2) relation between BDI score and social activities; and (3) stress on the relatives that was mainly dependent on both the disability of the patients and their loss of social activities, whereas depression played a minor role. CONCLUSION: A high percentage of patients have depressive disorders 3 years or more after the stroke, independent of the side. Such mood disorders worsen the relationship between the disabled patients and their relatives and worsen leisure independent of the affected hemisphere. PMID- 9228893 TI - Exaggerated TSH responses to TRH in depressed patients with "normal" baseline TSH. AB - BACKGROUND: Subclinical hypothyroidism (elevated thyroid-stimulating hormone [TSH] with normal thyroid hormone levels) can present with depression. This may be confirmed by an exaggerated TSH response to thyrotropin-releasing hormone (TRH) on the TRH stimulation test (TRH-ST). The objective of this study was to determine the prevalence of exaggerated TRH-ST results in a sample of depressed patients with "high-normal" screening TSH levels. METHOD: Depressed patients with TSH levels of 3.00-5.50 mIU/L underwent a TRH-ST. After baseline TSH was drawn, TRH 400 micrograms was injected intravenously, and TSH samples were drawn at +20 min, +30 min, and +40 min postinjection. A rise in TSH after TRH (peak value minus baseline) of > 25 mIU/L represented an exaggerated TSH response. RESULTS: Twenty-three (38%) of 60 patients had an exaggerated TSH response to TRH. The 38% prevalence is significantly (Chi 2 = 59.65, df = 1, p < .001) greater than the 6% prevalence of positive TRH-ST results reported in the euthyroid general population. The prevalence of positive TRH-ST results was not attributable to differential patterns of psychotropic or thyroid hormone treatment. Unexpected observations were a lack of correlation in TSH levels week to week (r = .17, N.S.) and a lack of correlation between screening TSH value and subsequent TRH-ST results (r = .28, N.S.). CONCLUSION: Subtle thyroid underfunction may be contributing to depression in some patients with TSH in the upper half of the range usually considered normal. If so, then the TRH-ST may be more sensitive in identifying this than measurement of TSH alone. PMID- 9228894 TI - Safety in overdose of mirtazapine: a case report. PMID- 9228895 TI - Lamotrigine treatment of refractory bipolar disorder. PMID- 9228896 TI - Comparison of valproic acid and lithium in mania. PMID- 9228897 TI - Refractory depression, cardiovascular risk factors, and leukoariosis. PMID- 9228898 TI - Reversible parkinsonism in a 90-year-old man taking sertraline. PMID- 9228899 TI - Cognitive-behavioral management of drug-resistant major depressive disorder. AB - BACKGROUND: The application of cognitive-behavioral treatment to drug-resistant major depression has received little research attention. METHOD: Nineteen patients who failed to respond to at least two trials of antidepressant drugs of adequate dosages and duration were treated by cognitive-behavioral methods in an open trial. RESULTS: Three patients dropped out of treatment. The remaining 16 patients displayed a significant (p < .001) decrease in scores on the Clinical Interview for Depression after therapy. Twelve patients were judged to be in remission at the end of the trial; only 1 of these patients was found to have relapsed at a 2-year follow-up. Antidepressant drugs were discontinued in 8 of the 12 patients who responded to cognitive-behavioral treatment. CONCLUSION: These preliminary results suggest that a trial of cognitive-behavioral therapy by an experienced therapist should be performed before labeling an episode of major depression as "refractory" or "treatment resistant." These latter terms should apply only when a psychotherapeutic effort has been made. Until then, it seems more appropriate to define depression as "drug refractory" or "drug treatment resistant." PMID- 9228900 TI - Integrating faith and illness into practice. PMID- 9228901 TI - Manganese toxicity. PMID- 9228902 TI - Physician advice for problem alcohol drinkers. PMID- 9228903 TI - Influenza immunization for patients with MS. PMID- 9228904 TI - HRT for postmenopausal women. PMID- 9228905 TI - Misoprostol for cervical ripening and labor induction. PMID- 9228906 TI - Treatment of bleeding peptic ulcers with omeprazole. PMID- 9228907 TI - Coronary angioplasty vs thrombolysis in acute MI. PMID- 9228908 TI - Is amoxicillin beneficial in acute maxillary sinusitis? PMID- 9228909 TI - Levothyroxine bioequivalence. PMID- 9228910 TI - Clinical trials of interactive computerized patient education: implications for family practice. AB - A systematic review of randomized clinical trials was conducted to evaluate the acceptability and usefulness of computerized patient education interventions. The Columbia Registry, MEDLINE, Health, BIOSIS, and CINAHL bibliographic databases were searched. Selection was based on the following criteria: (1) randomized controlled clinical trials, (2) educational patient-computer interaction, and (3) effect measured on the process or outcome of care. Twenty-two studies met the selection criteria. Of these, 13 (59%) used instructional programs for educational intervention. Five studies (22.7%) tested information support networks, and four (18%) evaluated systems for health assessment and history taking. The most frequently targeted clinical application area was diabetes mellitus (n = 7). All studies, except one on the treatment of alcoholism, reported positive results for interactive educational intervention. All diabetes education studies, in particular, reported decreased blood glucose levels among patients exposed to this intervention. Computerized educational interventions can lead to improved health status in several major areas of care, and appear not to be a substitute for, but a valuable supplement to, face-to-face time with physicians. PMID- 9228911 TI - Alternative medicine. PMID- 9228912 TI - The right intentions: errors and accountability. PMID- 9228913 TI - Adverse events in primary care identified from a risk-management database. AB - BACKGROUND: The inevitability of adverse events in medicine arises from human fallibility, negligent care, limits of medical knowledge, risks inherent in medical practice, and biological variability among individuals. A better understanding of the nature and causes of adverse events is necessary to reduce their occurrence and limit their harm. This study describes adverse events identified from a risk-management database that occurred in an out-patient primary care setting. METHODS: Incident reports filed with the risk-management office of an academic medical center between January 1, 1991, and June 30, 1996, by eight primary health care clinics affiliated with the center were eligible for the study. Two independent reviewers assessed the incidents to determine whether there were adverse medical events. Incidents classified as adverse events were analyzed to determine the cause, potential preventability, and outcome. RESULTS: The prevalence of adverse events was 3.7 per 100,000 clinic visits over a period of 5 1/2 years. Twenty-nine of 35 (83%) adverse events were due to medical errors and were considered preventable. The causes of the adverse events included 9 diagnostic errors (26%), 11 treatment errors (31%), and 9 other errors (26%). Of the adverse events attributed to medical errors, 4 (14%) resulted in a permanent, disabling injury and 1 (3%) resulted in a death. CONCLUSIONS: Serious adverse events appear to occur infrequently in primary care outpatient practice, although these data probably underestimate the overall prevalence. To reduce or prevent the occurrence of adverse events in primary care, better systems for recognizing and tracking them and for assessing their causes are needed. PMID- 9228914 TI - Referrals of adult patients from primary care: demographic disparities and their relationship to HMO insurance. AB - BACKGROUND: Increasing enrollment in managed care organizations and dissatisfaction with policies to restrict direct access to specialists have intensified interest in referrals from primary care physicians to specialists. We examined the associations of demographic factors and insurance with referrals of adult patients by primary care physicians. METHODS: Office visits of adult patients to primary care physicians (general practitioners, family physicians, and internists) reported in the National Ambulatory Care Survey for the years 1985 through 1992 were used to examine referrals by primary care physicians. Regression analyses were adjusted for patient factors (age, sex, race, insurance, case mix, diagnostic category, new problem or not, new patient or not, and visit length), physician factors (age, sex, specialty, and degree of specialization), and practice factors (proportion of HMO patients, rural location, region, and study year). RESULTS: Overall, 4.5% of patients were referred compared with 7.5% of patients with HMO insurance. After adjustment, an increased likelihood of referral was associated with being a male patient, having fewer medications prescribed, not being seen before for the presenting problem, a longer visit, less physician specialization, seeing a female physician, seeing an internist, and seeing a physician with a greater proportion of patients with HMO insurance. Among patients with HMO insurance, no gender disparity in referral rate was observed, and patients who also had Medicaid or Medicare insurance were more likely to be referred. CONCLUSIONS: Male patients are more likely to be referred. HMO insurance may reduce this gender disparity and increase the access of patients with Medicaid and Medicare to specialty care. PMID- 9228915 TI - Prevalence and recognition of panic states in STARNET patients presenting with chest pain. AB - BACKGROUND: The purpose of this study was to document the prevalence of panic states in patients presenting with chest pain in primary care settings, to determine the recognition rate of panic states by family physicians, and to assess the impact of lack of recognition on interventions and costs. METHODS: Patients from the South Texas Ambulatory Research Network (STARNET) presenting with a new complaint of chest pain were asked to participate in the study. Before seeing their physician, subjects completed the panic disorder section of the Structured Clinical Interview (SCID) of the Diagnostic and Statistical Manual of Mental Disorders, Third Edition, Revised. The SCID was used to assign diagnoses of panic disorder, infrequent panic, or limited symptom attacks. Health care outcomes included medications prescribed, tests ordered, follow-up and referrals, costs, and physician diagnosis. RESULTS: Although approximately one half of the 51 patients in this study met criteria for either panic disorder or infrequent panic, few were recognized by physicians as having a panic state (kappa = -.003). Patients with panic disorder were more likely to receive follow-up or referral (P = .042), incurring higher follow-up costs (P = .080). Patients with infrequent panic received more testing (P = .008), with higher costs for testing (P = .001) and higher overall costs (P = .067). Panic-diagnosis associations were found between psychotropic (P = .001) and total (P = .070) medications as well as follow-up and referral costs (P = .009). CONCLUSIONS: Although common, panic states are rarely recognized in patients presenting with complaints of chest pain. The presence of panic leads to more testing, follow-up, and referral with subsequent higher costs. Failure to diagnose panic results in increased prescribing of medications, higher costs, and inappropriate pharmacotherapy. PMID- 9228916 TI - Measuring attributes of primary care: development of a new instrument. AB - BACKGROUND: The purpose of this study was to evaluate the measurement properties of an instrument developed to measure seven key aspects of the delivery of primary care from the perspective of patients visiting their family physician, and to report the association of these aspects with patient satisfaction. METHODS: A cross-sectional study design was used to examine the responses of 2899 patients visiting 138 family physicians' offices in Northeast Ohio. A 20-item research tool, the Components of Primary Care Index (CPCI), was created to measure the domains of primary care based on the new Institute of Medicine definition and on additional domains based on the literature. Patient satisfaction was measured with the Medical Outcomes Study 9-item visit rating form. The usual provider continuity (UPC) index was calculated as the proportion of visits to the index physician with relation to all physician visits for the past year by patient report. The CPCI was subjected to item and factor analysis. Scale scores were computed, and the association with patient satisfaction with the visit was tested by correlation. RESULTS: The factor analysis resulted in four stable and internally consistent factors. The factors were named: interpersonal communication, physician's accumulated knowledge of the patient, coordination of care, and patients' preference to see their regular physician. Each of the CPCI scale scores was significantly associated with patient satisfaction with the visit. The UPC index, length of time as a patient, and intensity of visits were not as strongly associated with the patient satisfaction measure. CONCLUSIONS: The CPCI provides a brief and reliable measure of four important aspects of the delivery of primary care. The components of primary care are associated with patient satisfaction with visits to family physicians. The CPCI could be used with other outcomes and to assess the effect of interventions and systems changes on the delivery of critical aspects of primary care. PMID- 9228917 TI - Patient knowledge of upper respiratory infections: implications for antibiotic expectations and unnecessary utilization. AB - BACKGROUND: Upper respiratory infections (URIs) account for many of the visits in primary care and are commonly treated with ineffective antibiotic therapy. The purpose of this study was to examine patient beliefs in the effectiveness of antibiotics and the likelihood of seeking care for normal presentations of URIs. METHODS: We conducted a survey of 961 adults (> or = 18 years of age) from an undifferentiated patient population in a university-based family practice residency clinic in metropolitan Kentucky, a private internal medicine practice in nonmetropolitan Kentucky, and, in metropolitan Louisiana, an emergency department and a convenience sample from the community. RESULTS: Seventy-two percent of the sample reported that they would seek care with a condition of 5 days' duration with cough, sore throat, and discolored nasal discharge. Sixty-one percent of the sample expressed their belief that antibiotics are effective for a condition of 5 days' duration with cough, sore throat, and clear nasal discharge; 79% said that they believed antibiotics are effective when there is discolored discharge (P = .0001). Medicaid recipients were most likely to seek care across the symptom complexes. Higher education was related to a decreased belief in the effectiveness of antibiotics for the scenario with clear discharge (P .001), but to an increased belief in the effectiveness of antibiotics in the scenario with discolored discharge (P = .003). The strongest predictor of both likelihood of utilization and belief in effectiveness of antibiotics was usual use of antibiotics for the URI symptom complexes. CONCLUSIONS: Patients lack understanding of the normal presentation of a URI and the effectiveness of antibiotics as a treatment. A confusion about the meaning of discolored nasal discharge is particularly evident, and past antibiotic use may contribute to inappropriate utilization and expectations for antibiotics. PMID- 9228918 TI - Striae in adolescents mistaken for physical abuse. AB - Physiological striae are common in adolescence, occurring in the lumbar and gluteal regions, the upper thighs, breast, lower abdomen, and back. The lesions may be mistaken for nonaccidental injury, that is, physical abuse. We present four cases of adolescents with lesions thought to be due to physical abuse. Three of these cases were revealed during a school screening program for scoliosis; of the 2600 adolescents screened, aged 12 to 16 years, 168 were found to have striae. One case was found by a family physician when a young boy presented with low back pain. Since striae may be mistakenly ascribed to physical abuse, it is important for family physicians, nurses, and pediatricians to be familiar with this benign condition. PMID- 9228919 TI - Monocyte chemoattractant protein-1 mRNA expression in the human burn wound. AB - The inflammatory response following a thermal insult begins with the skin itself. Langerhan's cells, tissue macrophages, keratinocytes, fibroblasts, and endothelial cells contribute to the initial events of wound healing with active and passive release of cell mediators. One of the mediators potentially important to the repair process is monocyte chemoattractant protein-1 (MCP-1). Macrophages, fibroblasts, endothelial cells, and keratinocytes can produce MCP-1 in response to inflammatory stimuli. Therefore, we evaluated 10 human burn wound specimens for MCP-1 mRNA using in situ hybridization. Selected specimens of different ages were examined using combined in situ hybridization and immunocytochemistry to identify cell types that expressed MCP-1 mRNA. Antibodies to HAM56 for macrophages, CD45 for bone marrow-derived cells, Factor VIII for endothelial cells, and Factor XIIIa for dermal antigen-presenting cells were included in these experiments. By Postburn Day 2, basal layer keratinocytes at the edges of the wound had upregulated MCP-1 message; the increased signal persisted in the rate pegs deep in the dermal wound bed through 49 days postinjury. Occasional FXIIIa+ immunostained dermal cells expressed MCP-1 mRNA. Islands of granulation tissue throughout the wound bed were positive for increased expression of MCP-1; endothelial cells and inflammatory cells both contributed to this upregulated signal. Our data support the theory that the skin itself is a component of the immune system and that noninflammatory cells contribute to the initiation and maintenance of the inflammation at a wound site. Failure to produce MCP-1 or other related mediators by indigenous cutaneous cells may delay the inflammatory response to injury and potentially disrupt other essential phases of wound repair. PMID- 9228920 TI - Adenoviral thymidine kinase prodrug gene therapy inhibits sarcoma growth in vivo. AB - Local recurrence of sarcoma is due to residual tumor cells remaining after surgical resection and is associated with decreased survival. We implemented adenoviral-mediated transfer of the herpes simplex thymidine kinase (HSTK) gene with subsequent ganciclovir (GCV) administration to treat a model of residual sarcoma, [3H]Thymidine uptake in MCA sarcoma cells was determined after infection with replication incompetent adenovirus of the AdMLP.HSTK construct in the presence of GCV. In vivo efficacy was evaluated in a model of residual sarcoma when 9 mg of MCA tumor was implanted into the latissimus muscle of Fischer 344 rats. Three days after implantation, animals were randomized to receive AdMLP.HSTK, AdCMV. Null, or viral suspension buffer intratumorally. From Day 4, animals were administered b.i.d. GCV (50 mg/kg) or saline ip. Tumors were excised on Day 14 and weighed. Statistical analysis was by Mann-Whitney U test. In vitro: [3H]thymidine incorporation was significantly decreased in MCA sarcoma cells infected with AdMLP.HSTK in the presence of GCV (P < 0.05). In vivo: Growth of MCA sarcoma treated with AdMLP.HSTK and GCV was significantly inhibited. Final tumor weights in the AdMLP.HSTK/GCV group were lower than all control groups (P < 0.05). A significant antitumor growth effect on MCA sarcoma was seen with adenoviral-mediated transfer of the HSTK gene and GCV administration, both in vitro and in an in vivo model of residual disease. This prodrug gene therapy strategy warrants investigation as an adjuvant modality in the management of sarcoma. PMID- 9228921 TI - Sinusoidal flow block after warm ischemia in rats with diet-induced fatty liver. AB - Donor livers with massive fatty infiltration reportedly are susceptible to ischemia/reperfusion injury after transplantation, which contributes to risk of primary nonfunction. We investigated the effect of warm ischemia and reperfusion on sinusoidal microcirculation in rats with fatty livers from a choline-deficient diet. Rats were subjected to partial hepatic warm ischemia for 30, 60, or 90 min. In a second study, an anti-ICAM-1 monoclonal antibody was injected intraportally 2 min after a 60-min ischemic period. In both studies, injury was assessed by liver histology 6 hr after vascular clamp release and by animal survival. After 30 min of hepatic warm ischemia, almost all control and fatty-liver rats survived 7 days. After 60-min ischemia, however, survival was significantly less in rats with fatty livers than in controls with normal livers (10% vs 90%, P < 0.0001). Histologically, rats with fatty livers showed marked sinusoidal congestion, especially in the midzone of the acinus, while control rats showed no disturbance in microcirculation. In rats with fatty livers treated with intraportal injection of an anti-ICAM-1 antibody, sinusoidal microcirculation was well preserved and the 7-day survival rate after warm ischemia was improved (50% vs no antibody 10%; P = 0.0112). In fatty livers, midzonal sinusoidal flow block occurs after hepatic warm ischemia and reperfusion. Although intraportal injection of an anti-ICAM-1 monoclonal antibody corrected this microcirculatory failure, animal survival was not as good as for controls without fatty livers. These results suggest that both sinusoidal microcirculatory failure and ischemic hepatocellular damage contribute to warm ischemia/reperfusion injury in fatty livers. PMID- 9228922 TI - IL-6 release after intestinal ischemia/reperfusion in rats is under partial control of TNF. AB - Although there is much evidence to substantiate the view that tumor necrosis factor (TNF) plays a pivotal role in the pathogenesis of multiple organ injury subsequent to intestinal ischemia/reperfusion (I/R), it is still unclear whether TNF is involved in triggering the release of other inflammatory mediators in this condition. The current study was designed to determine the potential effects of TNF blockade, by means of monoclonal antibody (TNF MoAb) treatment, on plasma interleukin 6 (IL-6) in rats after acute intestinal I/R injury. Anesthetized rats underwent 75-min occlusion of superior mesenteric artery followed by 6 hr of reperfusion. The animals were treated with TNF MoAb or control protein at a dose of 20 mg/kg i.v. 30 min before the onset of I/R. Similar IL-6 responses in both the portal and the systemic circulation were observed in animals subjected to intestinal I/R, who showed a progressive increase in plasma IL-6 concentration upon release of the clamp. In animals receiving TNF MoAb before I/R, the subsequent IL-6 release following reperfusion was significantly blunted compared to the levels in controls (P < 0.05). The present study demonstrates that the activation and/or release of IL-6 in the setting of acute gut I/R may, at least in part, be mediated via TNF-dependent mechanisms, providing further evidence that a complex interaction exists between TNF and IL-6. PMID- 9228923 TI - Binding and internalization of anti-sarcoma IgG and IgM antibodies. AB - A variety of monoclonal antibodies directed against tumor cell surface antigens have been employed for tumor detection and delivery of toxins and radioisotopes to tumors in situ. The sequence of events following antibody binding to tumor cells may be critical to the success or failure of tumor imaging or therapy. Using indirect immunofluorescence, we have examined the binding and internalization of two different monoclonal antibodies (19-24 and MFH 4.10) by HT 1080, a cultured human sarcoma cell line. MAb 19-24 (IgG1) and MAb MFH 4.10 (IgM) are directed against surface antigens on cells from human malignant fibrous histiocytoma (MFH). For both of these antibodies, the initial binding on the surface of HT-1080 cells at 4 degrees C was uniform. After binding secondary fluorescent antibody, monoclonal antibodies underwent rapid internalization at 37 degrees C. Surface-bound antigen-antibody complexes were completely cleared from the cell surface in 30-60 min. In the absence of secondary fluorescent antibody, MAb 19-24 remained bound to the cell surface at 37 degrees C for up to 20 hr without being released or internalized. However, under these same conditions MAb MFH 4.10 was internalized completely within 30 min by HT-1080 cells and did not subsequently return to the cell surface. Such differences in the mechanisms of binding and uptake for these two antibodies may have significant implications for their future clinical or therapeutic uses. PMID- 9228924 TI - Increased immune responses to regenerating partial liver grafts in the rat. AB - The relationship between liver regeneration and the induction of the immune response is uncertain. We hypothesize that the altered environment of the regenerating liver allograft increases the immune response to the allograft. In DA (RT1a) to LEW (RT1I) rats, hepatectomized, small-for-size and whole, normal for-size liver transplants were performed. Naive and 70% hepatectomized LEW served as controls. Animals were assessed for survival, mass restoration, and host alloresponses. Although 30% partial allografts regenerated well to achieve a volume nearly equal to that of recipient's native liver in 7 days, survival was significantly shorter than that of the recipients of whole grafts (8.8 +/- 0.4 vs 10.3 +/- 1.2 days, n = 6, P < 0.02). When compared on Day 4 after transplantation, histologic examination revealed a more vigorous cellular infiltration in the sinusoidal area in the partial liver transplant group. Phenotypic analysis of thymocytes showed a predominance of more mature phenotypes in the partial group, including more prominent decrease in the frequency of CD4, CD8-double-positive cells and increase in that of alpha beta TCRhigh cells. Proliferative activity of thymocytes in response to Con A was higher in the partial group than in the whole group. MLR of splenocytes against donor-type antigens was higher in the partial group, whereas reactivity against third party was the same as in other groups. These data suggest that host cellular responses to the allograft are enhanced in the regenerating, small-for-size liver graft. These findings have implications in the clinical management of liver recipients with damaged or small for size livers. PMID- 9228925 TI - Constant delivery of proinsulin by encapsulation of transfected cells. AB - Gene therapy is a potentially excellent approach for the treatment of diabetes instead of pancreas and islet transplantation. However, one difficulty involved in gene therapy for diabetes is a control of insulin/proinsulin production by the cells transfected with insulin cDNA. The purpose of this study is to examine whether control of the proliferation of transfected cells by encapsulation is a feasible approach for the constant delivery of proinsulin to avoid a life threatening hypoglycemic state. Previously, we established a mouse fibroblast Ltk cells transfected with human insulin cDNA producing human proinsulin (91 ng/24 hr/10(6) cells). These cells were encapsulated with semipermeable 5% agarose gel and proinsulin production was examined by in vitro long-term culture system. Intraperitoneal implantation into streptozocin (STZ)-induced diabetic mice was performed to investigate in vivo function of the encapsulated cells. The data from the in vitro study demonstrated that encapsulation of 2 x 10(6) transfectants enabled the stable production of proinsulin for over 80 days (204.4 +/- 5.18 ng/ml/day). Implantation of the encapsulated 2 x 10(7) transfectants improved the hyperglycemic state of diabetic mice for 30 days on the mean value of blood glucose concentration (n = 20). Histological analysis revealed pericapsular inflammation at 30 days after implantation and this may result in malfunction of encapsulated cells. Constant production and delivery of proinsulin could be achieved by encapsulating the human insulin cDNA-transfected cells using 5% agarose. Control of the proliferation of transfected cells appears to be an important factor for constant delivery of human proinsulin toward gene therapy of diabetes mellitus. PMID- 9228926 TI - Biochemical markers of upper esophageal sphincter compliance in patients with Zenker's diverticulum. AB - The aim of this study was to investigate the biochemical basis of biomechanical and morphological alterations of upper esophageal sphincter, which have been reported in patients with Zenker's diverticulum. 4-L-Hydroxyproline (4-Hyp) (collagen), isodesmosine (Ides), and desmosine (Des) (elastin) contents were measured in samples of cricopharyngeal muscle (CPM) and muscularis propria of the esophagus below the CPM. The specimens were collected from seven patients operated for Zenker's diverticulum and eight cadavers, without esophageal and connective tissue disease, 4-Hyp was assayed colorimetrically, Ides and Des by high-performance liquid chromatography. Mean (+/-SEM) values were compared by Mann-Whitney U test. In patients, collagen content was significantly increased, both in CPM and in the muscularis propria of the esophagus below the CPM (P < 0.05). In CPM, Ides to Des and collagen to elastin ratios were significantly higher in patients than in controls (P < 0.05). Both the CPM and the upper muscular cuff of the esophagus appear to be involved in the pathogenesis of Zenker's diverticulum. This finding supports the extension of the myotomy to the muscularis propria of the esophagus below the CPM. The alterated Ides to Des ratio suggests a primary disease of CPM as a cause of Zenker's diverticulum. PMID- 9228927 TI - Lipopolysaccharide-mediated hepatic glutathione depletion and progressive mitochondrial damage in mice: protective effect of glutathione monoethyl ester. AB - Overproduction of reactive oxygen intermediates (ROI) may have an important role in the pathophysiology of lipopolysaccharide-mediated liver-injury. This study examined the role of cytosolic and mitochondrial glutathione in protecting hepatocytes from oxidative stress during exposure to lipopolysaccharide. In addition, the possible participation of changes of inner mitochondrial membrane permeability in lipopolysaccharide-induced hepatotoxicity was investigated. The changes of hepatic glutathione content following lipopolysaccharide challenge (2 mg/kg) were measured in mice by reverse-phase high-performance liquid chromatography. Glutathione depletion and a glutathione-rich state were produced by intraperitoneal administration of a specific inhibitor of gamma-glutamyl cysteine synthetase, buthionine sulfoximine (3 mmol/kg), and by administration of glutathione monoethyl ester (10 mmol/kg), respectively. Intracellular ROI generation and the mitochondrial membrane potential were quantified by flow cytometry. Changes of inner mitochondrial membrane permeability in hepatocytes were assessed by radioactive sucrose entrapment. There was increased production of ROI along with depletion of cellular and mitochondrial glutathione in the liver after lipopolysaccharide administration. There was also a change of inner mitochondrial membrane permeability in hepatocytes, with the loss of coupled functions. Buthionine sulfoximine decreased the hepatic antioxidant capacity, worsened mitochondrial function, and reduced the survival rate of the mice. In contrast, glutathione monoethyl ester improved all of these parameters. Glutathione may have an important role in cellular defenses against lipopolysaccharide-induced liver damage in mice, and excessive oxidative stress may precipitate the mitochondrial membrane permeability transition in hepatocytes and lead to cell death. PMID- 9228928 TI - Mechanism of intestinal mucosal immune dysfunction following trauma-hemorrhage: increased apoptosis associated with elevated Fas expression in Peyer's patches. AB - Although intestinal mucosal immune dysfunction occurs after trauma and hemorrhage and appears to contribute to infectious complications, the mechanism is not known. In this regard, the inappropriate induction of immune cell apoptosis may contribute to the immune dysfunction following trauma and hemorrhage. To study this, we examined Peyer's patch cells for evidence of apoptosis and changes in Fas protein expression, as well as alterations in the relative lymphocyte subpopulations after trauma and hemorrhagic shock. Male C3H/HeN mice underwent sham operation, trauma (i.e., laparotomy), hemorrhagic shock (mean arterial blood pressure of 35 +/- 5 mmHg for 90 min, followed by adequate crystalloid resuscitation), or trauma plus hemorrhage. Peyer's patch cells were isolated at 24 and 72 hr after the procedure. The percentage of apoptotic cells, Fas protein expression, and cell percentage of phenotype were determined by flow cytometry. The results indicate that trauma alone induced no significant change in the measured parameters. However, (i) there were a significant decrease in total viable cell yield and an increased apoptosis in Peyer's patch cells at 24 and 72 hr following hemorrhage or trauma plus hemorrhage; (ii) the increased apoptosis in Peyer's patch was predominantly in cells of the B-cell lineage; (iii) the increased apoptosis was associated with an elevated Fas expression. In conclusion, hemorrhage alone or trauma plus hemorrhage can induce increased apoptosis in Peyer's patch cells, which is associated with the increased Fas expression. Thus, apoptotic involution may play an important role in the depression of intestinal mucosal immune function after trauma and hemorrhagic shock. PMID- 9228929 TI - Effect of nitroglycerin and cerebrospinal fluid drainage on spinal cord perfusion pressure and paraplegia during aortic cross-clamping. AB - When sodium nitroprusside (SNP) is used to control proximal blood pressure (Px BP) during cross-clamping (AXC) of the thoracic aorta, it decreases spinal cord perfusion pressure (SCPP) by reducing distal aortic pressure (Ds-BP) and increasing cerebrospinal fluid pressure (CSFP). The decrease cannot be reversed by CSF drainage (CSFD) because such drainage is limited by a reduction in compliance of the spinal canal. Nitroglycerin can also be used to control Px-BP, but its effect on CSF dynamics has not previously been investigated. In the present study we have compared the effects of NTG alone and in combination with CSFD, with SNP and CSFD. Each group (Gp) of six dogs was treated with SNP + CSFD (Gp 1), NTG alone (Gp 2), and NTG + CSFD (Gp 3). Left carotid and right femoral arteries were catheterized to monitor Px-BP and Ds-BP, respectively. CSFP was monitored and CSF was drained through a spinal needle placed in the cisterna cerebellomedullaris. The thoracic aorta was cross-clamped via a left thoracotomy for 60 min. Data were acquired at baseline, during aortic occlusion, and 24 hr after surgery. There were no significant differences in any measurements among the three groups before AXC; after AXC, Px-BP was maintained between 85 and 95 mm Hg in all groups. Ds-BP was significantly lower in Gp 1 than Gp 2 and 3 (7 +/- 2 mm Hg vs. 13 +/- 3 mm Hg and 17 +/- 2 mm Hg, respectively P < 0.05). CSFP did not differ between Gp 1 and 2 (10 +/- 3 mm Hg vs. 9 +/- 1 mm Hg, P > 0.05). CSFD effectively kept CSFP at zero values in Gp 3 during AXC, but not in Gp 1. SCPP was significantly higher in Gp 3 than in Gp 1 and 2 (17 +/- 2 mm Hg vs -3 +/- 4 mm Hg and 4 +/- 1 mm Hg, respectively, P < 0.05). All animals in Gp 1 and 2 suffered paraplegia, as opposed to none in Gp 3. NTG causes paraplegia by decreasing SCPP. When used in conjunction with CSFD, it controls Px-BP without causing paraplegia. CSFD cannot counteract the negative effects of SNP on SCPP; therefore, SNP contributes to postoperative paraplegia. The effects of NTG on cerebrospinal fluid dynamics are different from those of SNP. We caution surgeons against the use of NTG without CSFD during aortic cross-clamping. PMID- 9228930 TI - Impact of a problem-based learning conference on surgery residents' in training exam (ABSITE) scores. American Board of Surgery in Training Exam. AB - The impact of problem-based learning on surgery residents' education is unknown. In this study we measured the impact of a weekly structured problem-based learning conference on surgery residents' ABSITE scores and compared it to traditional clinical conferences and self-studying. A questionnaire was designed to determine the perceived quality of the basic (PQCB), and the clinical (PQCC) conferences as well as self-studying (PQCS). The Pearson correlation between PQCB, PQCC, PQCS, and attendance at the basic science conference and each of the ABSITE total score (ABSITE), basic science (BS) and clinical science (CS) component scores were calculated. PQCS (4.2) was significantly higher than PQCB (2.9) and PQCC (2.5) (P = 0.0002). PQCS and PQCB correlated highly with each of ABSITE, CS, and BS while PQCc did not show any correlation. A high correlation was also observed between attendance at basic science and each of ABSITE, CS, and BS but narrowly missed significance. It was also observed that BS scores highly correlated to the CS scores at all postgraduate levels (P = 0.0001). We conclude that performance on all components of the ABSITE is mostly dependent on individual residents. This individual factor is boosted by self-studying which can be motivated by instituting a problem-based learning technique within the program. Traditional conferences even if popular among residents have no impact on measurement tests. PMID- 9228931 TI - Gastric antigen challenge releases gastrin and eicosanoids and protects against ethanol. AB - Various gastrointestinal functions such as mucosal blood flow and mucus secretion can be influenced immunologically. Rats were systemically sensitized with 4 hydroxy-3-iodo-5-nitro-phenylacetic acid (NIP), a synthetic antigen. Mucosal release of gastrin, prostaglandin F2 alpha, 6-keto-prostaglandin F1 alpha, and leukotriene C4 was measured after intragastric or in vitro antigen challenge. Gastric protection from ethanol was determined. In sensitized rats, intragastric antigen challenge increased release of gastrin from the antral mucosa ex vivo and tended to increase release of prostaglandin F2 alpha. Likewise, antral mucosa of sensitized rats released significantly more gastrin and prostaglandin F2 alpha during in vitro antigen challenge than during incubation in the absence of antigen. Release of 6-keto-prostaglandin F1 alpha and leukotriene C4 was not affected by the immunologic reaction. Topical antigen challenge in sensitized rats reduced gastric mucosal damage caused by ethanol by 50%. The immunologically induced gastroprotection was significantly attenuated by pretreatment with indomethacin. The findings show that specific antigen challenge renders the gastric mucosa more resistant against the injurious effect of ethanol indicating that the stomach is a target organ of immunological reactions. As gastrin and prostaglandins exert potent protective effects, release of these mediators may contribute to the protective response to gastric mucosal immune activation. PMID- 9228932 TI - Vasoconstriction increases pulmonary nitric oxide synthesis and circulating cyclic GMP. AB - Vascular shear stress increases when blood flow or blood viscosity increases or when vessel diameter decreases. In the systemic circulation, shear stress is a potent stimulus for endothelial nitric oxide synthesis. We studied isolated rat lungs to determine whether increasing shear stress increases nitric oxide synthesis in the pulmonary circulation. Lungs were given the vasoconstrictor, U46619 (a thromboxane analogue), and perfused at constant flow rates or at constant pressure, since constant pressure perfusion minimizes changes in shear stress with vasoconstriction. The subsequent effect of the NOS inhibitor, N omega methyl-L-arginine (LMA), or the soluble guanylyl cyclase inhibitor, 6-anilino-5,8 quinolinodione (LY83583) was assessed. Changes in pulmonary vascular resistance (PVR), pulmonary vascular compliance, and perfusate cyclic GMP concentration were measured as indicators of nitric oxide synthesis. The effect of the cyclic GMP specific (type V) phosphodiesterase inhibitor, zaprinast, on perfusate cyclic GMP concentrations was also examined. An infusion of U46619 consistently increased PVR and decreased compliance. LMA and LY83583 also increased PVR in U46619 treated lungs perfused at constant flow rates, primarily by increasing precapillary resistance. LMA had no effect in U46619-treated lungs perfused at constant pressure. Perfusate cyclic GMP concentrations increased significantly after U46619 in lungs perfused at constant flow rates, but cyclic GMP levels did not change after U46619 in lungs perfused at constant pressure. Zaprinast also increased perfusate cyclic GMP, demonstrating that increases in intracellular cyclic GMP are reflected in circulating cyclic GMP concentrations. We conclude that vasoconstriction with U46619 increases nitric oxide synthesis in isolated rat lungs. Lungs perfused at constant pressure respond differently to NOS inhibitors compared to those perfused at constant flow, suggesting that shear stress may increase nitric oxide synthesis in the lung. Perfusate concentrations of cyclic GMP reflect activation of soluble guanylyl cyclase in this model. PMID- 9228933 TI - Altered wound arginine metabolism by corticosterone and retinoic acid. AB - Arginine metabolism plays an important role in many aspects of inflammation and wound healing. In this study, we tested the hypothesis that steroids and vitamin A have differential effects on arginine metabolism and thereby may provide a mechanism by which steroids impair wound healing, and vitamin A improves this impairment. Rats were treated with subcutaneous corticosterone pellets 2 days prior to wounding. Intraperitoneal injections of all-trans retinoic acid in peanut oil were administered at the same time and repeated 2 and 4 days later. Polyvinyl alcohol sponges were implanted subcutaneously through a dorsal incision. On Postwounding Days 1, 5, 10, and 15, wound fluid was recovered from the sponges and assayed for nitrite/nitrate (NOx), citrulline, arginine, and ornithine concentrations as well as arginase activity. Steroid treatment decreased the metabolism of arginine to nitric oxide in the early phase of wound healing, and retinoic acid did not change this relationship. Corticosterone also decreased metabolism of arginine to ornithine in the later wound. This depression was inhibited by concomitant administration of retinoic acid. Considering the importance of nitric oxide in host defense and ornithine as a precursor for polyamine and proline synthesis, these data provide a mechanism by which vitamin A improves wound strength, but does not improve wound infection rates in steroid treated animals. PMID- 9228934 TI - Effects of L-arginine supplementation on human lymphocyte proliferation in response to nonspecific and alloantigenic stimulation. AB - BACKGROUND: L-Arginine has been described as a potential immunostimulant in vitro and in vivo. Excessive arginine, however, may be counterproductive. Data support the concept of minimal arginine requirements for normal lymphocyte proliferation, but the results of supplementation with pharmacologic doses of arginine have been contradictory. We hypothesized that excessive arginine supplementation might result in a blunting of normal immune responses of human lymphocytes in vitro. MATERIALS AND METHODS: Peripheral blood mononuclear and T-cells were isolated from normal human donors. Cells were cultured in complete media with various concentrations of L-arginine, L-ornithine, and glycine. Lymphocytes were then stimulated with PHA or alloantigens, and proliferation was determined by measuring [3H]thymidine incorporation. RESULTS: Lymphocyte proliferation was inhibited by L-arginine at pharmacologic doses. The effects were completely reversible. This inhibition could not be prevented by lymphocyte stimulation with IL-2. Lymphocyte proliferation was more sensitive to inhibition by lower doses of arginine when alloantigens from irradiated fresh tumor cells or allogeneic lymphocytes were the stimuli. Finally, lymphocytes showed variable sensitivity to inhibition of proliferation in response to mitogen when treated with L-ornithine (little to no effect) or L-arginine (consistent inhibition at high doses). Pharmacologic doses of L-arginine result in reversible inhibition of normal lymphocyte proliferation in response to both mitogen and alloantigen. This inhibition could not be blocked by interleukin-2. CONCLUSIONS: We conclude that caution should be exercised when recommending aggressive L-arginine supplementation as a possible method to reverse clinical immunosuppression caused by cancer, malnutrition, or trauma. PMID- 9228935 TI - Biochemical assessment of cellular damage after adipocyte harvest. AB - Free fat transplantation for soft tissue augmentation yields variable results, which may be related to the technique of fat harvest. To compare the viability of adipocytes harvested by liposuction (sal) or by excision (exc), fat harvested by both techniques from seven lipectomy patients was analyzed by glycerol-3 phosphate dehydrogenase (G3PDH) enzyme assay. Leakage of this lipogenic enzyme through the plasma membrane is a potential indicator of fat cell damage. Preliminary experiments showed this assay to be sensitive and specific for adipocyte G3PDH activity. Treatment of fat tissue with collagenase H resulted in complete release of the component fat cells for analysis with less loss of G3PDH activity, compared to other collagenase preparations. Each sample was digested and separated into three compartments: mature adipocytes-floating layer (F), acellular supernatant (S), and stromal pellet (P). Samples from each compartment were assayed for G3PDH activity, normalized to DNA content, and represented as a percentage of the whole (F + S + P). Within the subgroups, the fat cell fraction of the liposuction samples (Fsal) showed statistically more activity than the excised samples (Fexc) by paired Student's t test (P = 0.004). The supernatant (representing leaked G3PDH) and pellet fractions of excised samples revealed more G3PDH activity than the same fractions from liposuctioned tissue; the former (Sexc) to a significant degree (P = 0.036). Using this assay, the results indicate that liposuction fat harvest does not result in increased fat cell damage compared to fat harvested by excision. PMID- 9228936 TI - Nice splice. PMID- 9228937 TI - The ion core in RNA folding. PMID- 9228938 TI - Protein folding and the Paracelsus challenge. PMID- 9228939 TI - EF-hands embrace. PMID- 9228940 TI - Bridging the gap. PMID- 9228941 TI - Picture story. Nice guys needn't finish last. PMID- 9228942 TI - A low energy short hydrogen bond in very high resolution structures of protein receptor--phosphate complexes. AB - A very short hydrogen bond between an Asp and a phosphate is established in two high resolution structures (0.98 and 1.05 A). A mutant complex that changes the Asp to an Asn, which forms a normal hydrogen bond, has a similar free energy of binding to the wild type complex, suggesting that the contribution of the short hydrogen bond is not extraordinarily strong. PMID- 9228943 TI - Gas access to the active site of Ni-Fe hydrogenases probed by X-ray crystallography and molecular dynamics. AB - The 2.54 A resolution structure of Ni-Fe hydrogenase has revealed the existence of hydrophobic channels connecting the molecular surface to the active site. A crystallographic analysis of xenon binding together with molecular dynamics simulations of xenon and H2 diffusion in the enzyme interior suggest that these channels serve as pathways for gas access to the active site. PMID- 9228944 TI - Solution structure of human CTLA-4 and delineation of a CD80/CD86 binding site conserved in CD28. AB - The structure of human CTLA-4 reveals that residues Met 99, Tyr 100 and Tyr 104 of the M99YPPPY104 motif are adjacent to a patch of charged surface residues on the A'GFCC' face of the protein. Mutation of these residues, which are conserved in the CTLA-4/CD28 family, significantly reduces binding to CD80 and/or CD86, implicating this patch as a ligand binding site. PMID- 9228945 TI - Structure of a calpain Ca(2+)-binding domain reveals a novel EF-hand and Ca(2+) induced conformational changes. AB - The crystal structure of a Ca(2+)-binding domain (dVI) of rat m-calpain has been determined at 2.3 A resolution, both with and without bound Ca2+. The structures reveal a unique fold incorporating five EF-hand motifs per monomer, three of which bind calcium at physiological calcium concentrations, with one showing a novel EF-hand coordination pattern. This investigation gives us a first view of the calcium-induced conformational changes, and consequently an insight into the mechanism of calcium induced activation in calpain. The crystal structures reveal a dVI homodimer which provides a preliminary model for the subunit dimerization in calpain. PMID- 9228946 TI - Crystal structure of calcium bound domain VI of calpain at 1.9 A resolution and its role in enzyme assembly, regulation, and inhibitor binding. AB - The three dimensional structure of calcium-bound domain VI of porcine calpain has been determined to 1.9 A resolution. The crystal structure reveals five EF-hands, one more than previously suggested. There are two EF-hand pairs, one pair (EF1 EF2) displays an 'open' conformation and the other (EF3-EF4) a 'closed' conformation. Unusually, a calcium atom is found at the C-terminal end of the calcium binding loop of EF4. With two additional residues in the calcium binding loop, the fifth EF-hand (EF5) is in a 'closed' conformation. EF5 pairs up with the corresponding fifth EF-hand of a non-crystallographically related molecule. Considering the EF5's role in a homodimer formation of domain VI, we suggest a model for the assembly of heterodimeric calpain. The crystal structure of a Ca2+ bound domain VI-inhibitor (PD150606) complex has been refined to 2.1 A resolution. A possible mode for calpain inhibition is discussed. PMID- 9228948 TI - A magnesium ion core at the heart of a ribozyme domain. AB - Large ribozymes require divalent metal ions to fold. We show here that the tertiary structure of the Tetrahymena group I intron P4-P6 domain nucleates around a magnesium ion core. In the domain crystal structure, five magnesium ions bind in a three-helix junction at the centre of the molecule. Single atom changes in any one of four magnesium sites in this three-helix junction destroy folding of the entire 160-nucleotide P4-P6 domain. The magnesium ion core may be the RNA counterpart to the protein hydrophobic core, burying parts of the RNA molecule in the native structure. PMID- 9228947 TI - Protein alchemy: changing beta-sheet into alpha-helix. AB - For most proteins the amino acid sequence determines the tertiary structure. The relative importance of the individual amino acids in specifying the fold, however, remains unclear. To highlight this, Creamer and Rose put forth the 'Paracelsus challenge': Design a protein with 50% sequence identity to a protein with a different fold. We have met this challenge by designing a sequence which retains 50% identity to a predominantly beta-sheet protein, but which now adopts a four helix bundle conformation and possesses the attributes of a native protein. Our results emphasize that a subset of the amino acid sequence is sufficient to specify a fold, and have implications both for structure prediction and design. PMID- 9228949 TI - The structure of a novel insecticidal neurotoxin, omega-atracotoxin-HV1, from the venom of an Australian funnel web spider. AB - A family of potent insecticidal toxins has recently been isolated from the venom of Australian funnel web spiders. Among these is the 37-residue peptide omega atracotoxin-HV1 (omega-ACTX-HV1) from Hadronyche versuta. We have chemically synthesized and folded omega-ACTX-HV1, shown that it is neurotoxic, ascertained its disulphide bonding pattern, and determined its three-dimensional solution structure using NMR spectroscopy. The structure consists of a solvent-accessible beta-hairpin protruding from a disulphide-bonded globular core comprising four beta-turns. The three intramolecular disulphide bonds from a cystine knot motif similar to that seen in several other neurotoxic peptides. Despite limited sequence identity, omega-ACTX-HV1 displays significant structural homology with the omega-agatoxins and omega-conotoxins, both of which are vertebrate calcium channel antagonists; however, in contrast with these toxins, we show that omega ACTX-HV1 inhibits insect, but not mammalian, voltage-gated calcium channel currents. PMID- 9228950 TI - Solution structure of the N-terminal zinc binding domain of HIV-1 integrase. AB - The solution structure of the N-terminal zinc binding domain (residues 1-55; IN1 55) of HIV-1 integrase has been solved by NMR spectroscopy. IN1-55 is dimeric, and each monomer comprises four helices with the zinc tetrahedrally coordinated to His 12, His 16, Cys 40 and Cys 43. IN1-55 exists in two interconverting conformational states that differ with regard to the coordination of the two histidine side chains to zinc. The different histidine arrangements are associated with large conformational differences in the polypeptide backbone (residues 9-18) around the coordinating histidines. The dimer interface is predominantly hydrophobic and is formed by the packing of the N-terminal end of helix 1, and helices 3 and 4. The monomer fold is remarkably similar to that of a number of helical DNA binding proteins containing a helix-turn-helix (HTH) motif with helices 2 and 3 of IN1-55 corresponding to the HTH motif. In contrast to the DNA binding proteins where the second helix of the HTH motif is employed for DNA recognition, IN1-55 uses this helix for dimerization. PMID- 9228951 TI - Crystal structure of tyrosine hydroxylase at 2.3 A and its implications for inherited neurodegenerative diseases. AB - Tyrosine hydroxylase (TyrOH) catalyzes the conversion of tyrosine to L-DOPA, the rate-limiting step in the biosynthesis of the catecholamines dopamine, adrenaline, and noradrenaline. TyrOH is highly homologous in terms of both protein sequence and catalytic mechanism to phenylalanine hydroxylase (PheOH) and tryptophan hydroxylase (TrpOH). The crystal structure of the catalytic and tetramerization domains of TyrOH reveals a novel alpha-helical basket holding the catalytic iron and a 40 A long anti-parallel coiled coil which forms the core of the tetramer. The catalytic iron is located 10 A below the enzyme surface in a 17 A deep active site pocket and is coordinated by the conserved residues His 331, His 336 and Glu 376. The structure provides a rationale for the effect of point mutations in TyrOH that cause L-DOPA responsive parkinsonism and Segawa's syndrome. The location of 112 different point mutations in PheOH that lead to phenylketonuria (PKU) are predicted based on the TyrOH structure. PMID- 9228952 TI - Crystal structure of the RAG1 dimerization domain reveals multiple zinc-binding motifs including a novel zinc binuclear cluster. AB - The crystal structure of the dimerization domain of the V(D)J recombination activating protein, RAG1, was solved using zinc anomalous scattering. The structure reveals an unusual combination of multi-class zinc-binding motifs, including a zinc RING finger and a C2H2 zinc finger, that together from a single structural domain. The domain also contains a unique zinc binuclear cluster in place of a normally mononuclear zinc site in the RING finger. Together, four zinc ions help organize the entire domain, including the two helices that form the dimer interface. PMID- 9228953 TI - Supervising gene therapy, openly. PMID- 9228954 TI - Employing cognitive behaviour therapy to reduce unemployment. PMID- 9228955 TI - Vindication of policy of vitamin A with measles vaccination. PMID- 9228956 TI - Why tyrosinase for treatment of melanoma. PMID- 9228957 TI - Predominance of type II fibres in exertional heat stroke. PMID- 9228958 TI - Abandoning empirical antibiotics for febrile children. PMID- 9228959 TI - Prevalence of wheeze and asthma and relation to atopy in urban and rural Ethiopia. AB - BACKGROUND: Asthma and allergy in developing countries may be associated with adoption of an urbanised "western" lifestyle. We compared the rates of asthma symptoms and atopy in urban populations in Jimma, southwest Ethiopia, at an early stage of economic development with those among the population of remote, rural, subsistence areas, and assessed the potential role of environmental aetiological factors leading to the differences. METHODS: Information on wheeze of 12 months' duration, diagnosed asthma, and cough for 3 months of the year was gathered by questionnaire in random household samples of 9844 people from urban Jimma and of 3032 from rural areas. Atopy was defined by allergen skin-test response to Dermatophagoides pteronyssinus and mixed threshings measured in a one-in-four subsample of those aged 5 years and older from both groups. FINDINGS: All respiratory symptoms were rare in children and were significantly less common overall in the rural than in urban group (wheeze odds ratio 0.31 [95% CI 0.22 0.43], p < 0.0001). Asthma was reported by 351 (3.6%) of the urban group, with a median reported duration of 8.5 years (IQR 4-17 years) that was unrelated to age. Atopy was a strong risk factor for asthma in urban Jimma. In the rural areas, skin sensitivity to mixed threshings was only slightly less common than in urban Jimma (0.67 [0.40-1.12], p = 0.13), whereas sensitivity to D pteronyssinus was significantly more common (3.24 [2.40-4.38], p < 0.0001), and since none of the 119 atopic individuals in the rural area reported wheeze or asthma, atopy was possibly associated with a reduction in the risk of disease among this group. Wheeze or D pteronyssinus sensitivity were positively associated with housing style, bedding materials, and use of malathion insecticide, but no single factor accounted for the urban-rural differences. INTERPRETATION: Wheeze and asthma are especially rare in rural subsistence areas, and atopy may be associated with a reduced prevalence of these symptoms in this environment. In urban Jimma, self reported asthma seemed to emerge as a clinical problem about 10 years before our study began, which is consistent with an effect of new environmental exposures. The factor or factors leading to the increase in asthma and allergy have not been identified, although exposures related to general changes in the domestic environment are likely to be involved. PMID- 9228960 TI - Radiation and genetic factors in the risk of second malignant neoplasms after a first cancer in childhood. AB - BACKGROUND: Radiotherapy and chemotherapy are associated with an increased risk of second malignant neoplasm (SMN). An association between SMN and familial aggregation has also been shown. The aim of this study was to investigate the role of familial factors in the risk of SMN and their potential interaction with the effect of treatment. METHODS: We devised a case-control study of 25 children with SMN (cases) and 96 children with no SMN after a cancer treatment (controls), taken from a cohort of 649 children treated at our institution between 1953 and 1985. A complete family history was obtained for patients and controls and a familial index defined to evaluate the degree of familial aggregation. The radiation dose given at 151 sites in the body was estimated for each radiotherapy course for each child. FINDINGS: Among family members of the 25 SMN cases, there were ten with early-onset (< or = 45 years) cancer, compared with eight among relatives of the 96 controls. Compared with patients who had no family history of early-onset cancer, those with one or more affected family members had an odds ratio for SMN of 4.7 (95% CI 1.3-17.1; p = 0.02). Adjustment for local radiation dose and exclusion of patients known to be predisposed to SMN (carriers of p53 mutation and those with Recklinghausen's disease) did not affect this risk substantially. INTERPRETATION: Both genetic factors and exposure to ionising radiation have independent effects on the risk of SMN. Follow-up of children treated for cancer should be especially vigilant when there is a family history of early-onset cancer. PMID- 9228961 TI - Effect of cognitive-behavioural training on job-finding among long-term unemployed people. AB - BACKGROUND: The principles of cognitive-behavioural therapy (CBT) have been applied successfully through individual psychotherapy to several psychiatric disorders. We adapted these principles to create a group--training programme for a non-psychiatric group-long-term (> 12 months) unemployed people. The aim was to investigate the effects of the programme on measures of mental health, job seeking, and job-finding. METHODS: 289 volunteers (of standard occupational classification professional groups) were randomly assigned to a CBT or control programme, matched for all variables other than specific content, that emphasised social support. 244 (134 CBT, 110 control) people started the programmes and 199 (109 CBT, 90 control) completed the whole 7 weeks of weekly 3 h sessions (including three CBT, seven control participants who withdrew because they obtained employment or full-time training). Questionnaires completed before training, on completion, and 3-4 months later (follow-up data available for 94 CBT, 89 control) assessed mental health, job-seeking activities, and success in job-finding. Analyses were based on those who completed the programmes. Participants were not aware that two interventions were being used. Investigators were aware of group allocation, but were accompanied in all programmes by co trainers who were non-investigators. FINDINGS: Before training, 80 (59%) CBT group participants and 59 (54%) controls scored 5 or more on the general health questionnaire (GHQ; taken to define psychiatric caseness). After training, 29 (21%) and 25 (23%), respectively, scored 5 or more (p < 0.001 for both decreases). Improvements in mean scores with training on the GHQ (between-group difference 3.91, p = 0.05) and in other measures of mental health were significantly greater in the CBT group than in the control group. There were no significant differences between the groups in job-seeking activity during or after training, but significantly more of the CBT group than of the control group had been successful in finding full-time work (38 [34%] vs 13 [13%], p < 0.001), by 4 months after completion of training. INTERPRETATION: These results suggest that group CBT training can improve mental health and produce tangible benefits in job-finding. Application of CBT among the unemployed is likely to benefit both individuals and society in general. PMID- 9228962 TI - Randomised trial of effect of vitamin A supplementation on antibody response to measles vaccine in Guinea-Bissau, west Africa. AB - BACKGROUND: WHO has recommended vitamin A supplementation for children aged 6 months or older in developing countries at the same time as immunisation. One study has reported significantly lower seroconversion ratios among children who have received vitamin A supplements with measles vaccine at age 6 months. The aim of our study was to assess the effect of vitamin A supplementation on antibody response to measles vaccination at age 9 months, which is the more common age for immunisation in developing countries. METHODS: In an urban community in Guinea Bissau, we did a randomised, double-blind, placebo-controlled study of the effect of simultaneous vaccination and vitamin A supplementation in 462 children who received either a two-dose schedule of measles vaccine at the ages of 6 months and 9 months (150 infants) or one dose of measles vaccine at age 9 months (312 infants). Children were followed up to the age of 18 months and a blood sample was then collected to assess the antibody response. FINDINGS: 397 (86%) of the children took part in the follow-up (52 [11%] had moved and 13 [3%] had died). Among children who received a two-dose vaccine schedule, seroconversion was 98%. There was no difference in seroconversion or geometric mean titre (GMT) for children receiving vitamin A compared with children receiving no supplement. Among children receiving only one dose of measles vaccine at age 9 months, seroconversion was 95%. The GMT was significantly higher in children receiving vitamin A than in those receiving no supplement (3704 vs 2439 mIU; GMT ratio 1.52 [1.22-1.88]). The effect on plasma antibody concentration in the blood was stronger for boys (3902 vs 1916 mIU; GMT ratio 2.04 [1.53-2.72]) than for girls (3502 vs 3017 mIU; GMT ratio 1.16 [0.85-1.58]) who had received vitamin A with measles vaccine. In a multivariate analysis of variance adjusted for sex, vitamin A supplementation was associated with higher antibody titres (p < 0.001). There was a significant interaction between vitamin A supplementation and sex (p = 0.02). INTERPRETATION: There is no indication that simultaneous administration of measles vaccine and vitamin A supplements has a negative effect on measles immunity. Among the children who had received two doses of measles vaccine at the ages of 6 months and 9 months, supplements of vitamin A had no significant effect. Among children only receiving one dose of measles vaccine at age 9 months, 100,000 IU vitamin A increased antibody concentrations, especially for boys. PMID- 9228963 TI - Autoimmune enteropathy and villous atrophy in adults. AB - BACKGROUND: Autoimmune enteropathy is a condition described in children and characterised by villous atrophy, which is unresponsive to any dietary restrictions, and by the presence of enterocyte autoantibodies. We report two adult patients who fulfilled all the criteria for the diagnosis of this disorder. METHODS: Over the past 5 years we have seen four adult patients (all women, median age 51.5 [range 38-64] years) with subtotal villous atrophy, which was unresponsive to a gluten-free diet. The patients were HLA-DQ2 positive. IgA antigliadin and antiendomysial antibodies were not found in any of the patients. We did an indirect immunofluorescence search for enterocyte autoantibodies on monkey jejunum and for other autoantibodies for all four patients. FINDINGS: Of the four patients, two were positive for enterocyte autoantibodies and one of these two patients was positive for antiactin, antiparietal cell, and antithyroid microsomal autoantibodies. INTERPRETATION: To the best of our knowledge the two patients affected by severe enteropathy, who had never responded to any exclusion diet, and who were positive for enterocyte autoantibodies are the first cases of autoimmune enteropathy described in adults. We propose that adult patients whose disorders are unresponsive to a gluten-free diet should be tested for enterocyte autoantibodies. PMID- 9228965 TI - Should we treat acute respiratory distress syndrome with inhaled nitric oxide? PMID- 9228964 TI - An extra-ordinary cause of megacolon. PMID- 9228966 TI - Trimethoprim-induced aseptic meningitis and uveitis. PMID- 9228967 TI - A new tick-transmitted disease due to Rickettsia slovaca. PMID- 9228968 TI - Intracoronary-stent placement for coronary artery disease. PMID- 9228969 TI - Morbidity from hepatitis B after introduction of nationwide immunisation in Italy. PMID- 9228970 TI - Atopic dermatitis in a child due to cow dander. PMID- 9228971 TI - Regression of mucosa-associated lymphoid-tissue lymphoma of rectum after eradication of Helicobacter pylori. PMID- 9228972 TI - Is complete spermiogenesis failure a good indication for spermatid conception? PMID- 9228973 TI - Effects of increased cholinergic activity on naming in aphasia. PMID- 9228974 TI - New BRCA2 mutation in an Ashkenazi Jewish family with breast and ovarian cancer. PMID- 9228975 TI - Increased apoptosis in the contralateral testis in patients with testicular torsion. PMID- 9228976 TI - Hypertrophic cardiomyopathy. PMID- 9228977 TI - Conformational disease. AB - Several diverse disorders, including the prevalent dementias and encephalopathies, are now believed to arise from the same general disease mechanism. In each, there is abnormal unfolding and then aggregation of an underlying protein. The gradual accumulation of these aggregates and the acceleration of their formation by stress explain the characteristic late or episodic onset of the clinical disease. The understanding of these processes at the molecular level is opening prospects of more rational approaches to investigation and therapy. PMID- 9228978 TI - The art of visual neglect. PMID- 9228979 TI - Global burden of disease. PMID- 9228980 TI - Global burden of disease. PMID- 9228981 TI - Global burden of disease. PMID- 9228982 TI - Global burden of disease. PMID- 9228983 TI - Global burden of disease. PMID- 9228984 TI - Global burden of disease. PMID- 9228985 TI - Global burden of disease. PMID- 9228986 TI - Global burden of disease. PMID- 9228987 TI - Microorganisms in chronic arthritis. PMID- 9228988 TI - Low-protein diet and progression of chronic renal failure. PMID- 9228989 TI - Low-protein diet and progression of chronic renal failure. PMID- 9228990 TI - Low-protein diet and progression of chronic renal failure. PMID- 9228991 TI - Vancomycin-resistant enterococci in vegetarians. PMID- 9228992 TI - Ibopamine and survival in severe congestive heart failure: PRIME II. PMID- 9228994 TI - How important a factor is social class in preterm birth? PMID- 9228993 TI - Ibopamine and survival in severe congestive heart failure: PRIME II. PMID- 9228995 TI - Tuberculosis: old lessons unlearnt? PMID- 9228996 TI - Surgery for cancer. PMID- 9228997 TI - More growth-chart confusion. PMID- 9228998 TI - Inclusion of women in clinical trials of antiretroviral agents for HIV/AIDS during pregnancy. PMID- 9229000 TI - Influence of cell cycle on HIV-1 expression differs among various models of chronic infection. AB - Because of an inherent dependence on host cell second and third messenger signaling pathways for activation of HIV-1 expression, a potential exists for a relationship between the induction of latent HIV-1 and cell-cycle-related events. To investigate this potential relationship, cellular models of latent HIV-1 infection (OM-10.1 promyelocytes, ACH-2 T-lymphocytes, and U1 promonocytes) were chemically treated or gamma-irradiated to synchronize cultures at each cell cycle stage and then examined for constitutive and TNF-alpha-induced HIV-1 expression. Cell cycle synchronization alone had no effect on HIV-1 expression in OM-10.1 and U1 cultures; whereas enhanced constitutive HIV-1 expression was observed in ACH-2 cultures at G2 + M. A 2 hour TNF-alpha treatment of all synchronized OM-10.1 cultures activated HIV-1 expression to a similar extent as unsynchronized cultures. In contrast, the extent of TNF-alpha-induced HIV-1 expression in ACH-2 S and G2 + M cultures and in the U1 G0/G1 culture was greater than that in unsynchronized control cultures. However, no delay in the initial response was observed. Thus, the influence of cell cycle on constitutive and induced HIV-1 expression varied in each cellular model and, therefore, may further relate to the different molecular mechanisms maintaining viral latency. PMID- 9228999 TI - Cross-reactive and group-specific immune responses to a neutralizing epitope of the human respiratory syncytial virus fusion protein. AB - Immune responses to a synthetic peptide corresponding to amino-acids 205-225 of the fusion protein from group B respiratory syncytial (RS) virus, were studied in mice and rabbits, and compared to a similar peptide from group A RS virus. Peptide 205-225 (B) was recognized by monoclonal antibody RS-348, and was immunogenic in both mice and rabbits, as was peptide 205-225 from the fusion protein of a group A strain. Peptide 205-225 (B) induced a proliferative T-cell response, demonstrating the existence of a T-cell epitope in this region of the fusion protein of group B viruses. Both peptides were able to induce a T-cell cross-reactive proliferation when mice were primed with either the homologous or the heterologous peptide. ELISA were performed using synthetic peptides or whole virus (from group A and B) as antigens. Mice anti-peptide sera recognized both homologous and heterologous peptides. A similar pattern was observed with RS virus strains. In indirect immunofluorescence assays, both anti-peptide rabbit sera recognized human nasal epithelial cells infected with A or B strains of RS virus. In contrast, while anti-peptide 205-225 rabbit serum from group A neutralized group A and B strains of RS virus, anti-peptide 205-225 rabbit serum from group B was unable to neutralize a group A virus, although it neutralized a group B strain. These results are similar to the immune response observed in children following primary RS virus infection. PMID- 9229001 TI - The coat protein gene of grapevine leafroll associated closterovirus-3: cloning, nucleotide sequencing and expression in transgenic plants. AB - A lambda ZAP II cDNA library was constructed by cloning cDNA prepared from a high molecular weight double-stranded RNA (dsRNA, ca. 18 kb) isolated from grapevine leafroll associated closterovirus-3 (GLRaV-3) infected tissues. This cDNA library was immuno-screened with GLRaV-3 coat protein specific polyclonal and monoclonal antibodies and three immuno-positive clones were identified. Analysis of nucleotide sequences from these clones revealed an open reading frame (ORF) which was truncated at the 3' end; the remainder of this ORF was obtained by sequencing a fourth clone that overlapped with one of the immunopositive clones. A total of 2028 bp was sequenced. The putative GLRaV-3 coat protein ORF, 939 bp, encodes a protein (referred to as p35) with a calculated M(r) of 34866. Multiple alignment of the p35 amino acid sequence with coat protein sequences from other closteroviruses revealed that the consensus amino acid residues (R and D) of filamentous plant viruses are preserved in the expected locations. The GLRaV-3 coat protein gene was then engineered for sense and antisense expression in transgenic plants. Transgenic Nicotiana benthamiana plants that contain the sense GLRaV-3 coat protein gene produced a 35 kDa protein that reacted with GLRaV-3 antibody in Western blot. PMID- 9229002 TI - Characterization of rice yellow mottle virus isolates in Sudano-Sahelian areas. AB - The use of a panel of polyclonal antisera and monoclonal antibodies (MAbs) raised against West African isolates of rice yellow mottle virus (RYMV) in ELISA resulted in separation of 73 RYMV isolates into three distinct serogroups. Using a set of differential rice varieties, the serogroups could be correlated to two RYMV pathotypes. A relationship was found between serological properties of the RYMV isolates and their probable ecological origin. It was concluded that RYMV isolates originating in closely related agroecological zones displayed variability in coat protein and pathogenicity. This should be taken into account in developing tolerant or resistant rice varieties. PMID- 9229003 TI - Genomic heterogeneity of small ruminant lentiviruses: existence of heterogeneous populations in sheep and of the same lentiviral genotypes in sheep and goats. AB - We have recently shown that French small ruminant lentiviruses (SRLV) from sheep are more similar to Caprine Arthritis Encephalitis Virus (CAEV) than to visna maedi virus (VMV) in a conserved region of the pol gene. To extend these results, we have examined sequences from a variable region of the env gene in French SRLV. We found that they were nearly equally distant from both CAEV and VMV strains, suggesting a considerable divergence since the initial introduction of the virus. Analysis of separate clones from individual animals showed that some carry a population of variant viruses. The study of further pol gene sequences from both goats and sheep suggests that viral variants show little or no host species specificity. A phylogenetic tree of pol gene sequences confirmed the presence of a novel genotype of SRLV in France. PMID- 9229004 TI - Mutations in the SIV env and the M13 lacZa gene generated in vitro by reverse transcriptases and DNA polymerases. AB - To investigate the accuracy of retroviral in vitro DNA replication we have examined with two fidelity assays the reverse transcriptases (RTs) from SIVagm, HIV-1, MoMLV as well for comparison the Klenow fragment from E. coli and DNA polymerase a from calf-thymus. These forward mutation assays measured the loss of bacteriophage M13 lacZa gene function by mutations. In the EnvlacZa assay frameshift mutations occurring during polymerisation of a 176 b long simian immunodeficiency virus (SIV) envelope (env) sequence were phenotypically detected by blue/white-plaque screening. To measure in addition substitutions, a 116 b long M13 lacZa gene DNA template was used as the mutational target (LacZa assay). With the SIVagm env gene DNA template, we observed similar levels of frameshift fidelity for all three RTs. Nevertheless, the SIVagm RT was slightly more accurate than the other RTs and nearly all frameshifts were observed at two homopolymeric runs of its homologous template. Measuring also substitution errors at the lacZa template the mutation frequency of the SIVagm RT increased 2.5 fold and that of the HIV-1 RT was enhanced by a factor of 3. PMID- 9229005 TI - The cell receptor level is reduced during persistent infection with influenza C virus. AB - Persistent influenza C virus infection of MDCK cells perpetuates the viral genome in a cell-associated form. Typically, virus production remains at a low level over extended periods, in the absence of lytic effects of replication. In this study, we demonstrate that persistently infected cells are very restricted in permissiveness for superinfection. By reconstitution experiments, using bovine brain gangliosides as artificial receptors, the degree of super-infection was markedly increased. Analysis of cellular receptor expression revealed reduced concentrations of sialoglycoproteins in general and a limited presentation of the major receptor gp40. Cocultures of persistently infected and uninfected cells (the latter carrying normal receptor levels) initiated a transient rise in virus titers. This kind of induction of virus synthesis appeared to be mainly receptor linked, since a receptor-deprived subline, MDCK II, did not give rise to a similar effect. Susceptibility of MDCK II cocultures could be partly restored by ganglioside treatment. In accordance to related virus systems, these findings on influenza C virus suggest a role of cell receptor concentrations in the regulation of long-term persistence. PMID- 9229006 TI - Genomic sequence of the RA27/3 vaccine strain of rubella virus. AB - The sequence of the genome of the RA27/3 vaccine strain of rubella virus (RUB) was determined. In the process, several discrepancies between the previously reported genomic sequences of two wild RUB strains (Therien and M33) were resolved. The genomes of all three strains contain 9762 nucleotides (nts), exclusive of the 3' poly A tract. In all three strains, the genome contains (5' to 3'), a 40 nt 5' untranslated region (UTR), an open reading frame (ORF) of 6348 nts that encodes nonstructural proteins, a 123 nt UTR between the two genomic ORFs, a 3189 nt ORF that encodes the structural proteins, and a 62 nt 3' UTR. The 5' end of the subgenomic RNA was found to correspond to a uridine residue at nt 6436 of the genomic RNA. At the nucleotide level, the sequence of the three strains varied by 1.0 to 2.8%, while at the amino acid level, the sequence varied by 1.1 to 2.4% over both ORFs. The RA27/3 sequence will be of use in identification of the determinants of its attenuation, in vaccine production control and in development of second generation RUB vaccines based on recombinant DNA technology. PMID- 9229007 TI - Definitive identification of louping ill virus by RT-PCR and sequencing in field populations of Ixodes ricinus on the Lochindorb estate. AB - Rapid and precise virus detection procedures are an important component of any epizootiological study. An automated one tube reverse transcriptase and nested primer polymerase chain reaction (RT-PCR) followed by nucleotide sequencing of the cDNA product, was used for the rapid detection and identification of louping ill (LI) virus in field caught Ixodes ricinus and compared with a classical isolation method i.e. infectivity in cell culture. The results establish the genetic identity of LI virus on the Lochindorb Estate. There was a high correlation between the results obtained by RT-PCR and infectivity assays. RT-PCR and sequencing proved to be a rapid and accurate system for identifying LI virus in field specimens. Development of this system should improve the capacity to undertake detailed epizootiological studies of LI virus. PMID- 9229008 TI - Identification, cloning and sequence analysis of the equine adenovirus 1 hexon gene. AB - Based on sequence homology with human adenovirus 2 (HAdV2), the hexon gene of equine adenovirus 1 (EAdV1) was identified. HindIII restriction fragments containing the hexon and other viral genes were cloned into the plasmids pUC19 and pBlueScript SK(-) and sequenced. The nucleotide sequence of the hexon gene was completely determined and partial sequence data were obtained for seven other EAdV1 genes. Amino acid (aa) sequence comparison with published adenovirus (AdV) proteins identified the genes for the IIIa, penton, pVII, PVI, 23K proteinase, DNA binding and 100K proteins. The eight EAdV1 genes appeared to be in the same relative order as homologous genes of other AdV. The EAdV1 hexon protein was encoded between the hexon-associated pVI upstream and the 23K proteinase gene downstream and comprised 2742 nucleotides which translated into 913 aa. Similar to other members of the genus Mastadenovirus the EAdV1 hexon yielded two highly conserved genome segments at the N- and C-termini which flanked intermediate variable and hypervariable regions. The majority of the residue differences between EAdV1 and other AdV hexons occurred in two loops that are known for other AdV to protrude from the surface of the nucleocapsid. Amino acid comparisons with other AdV hexons revealed highest homology with HAdV12 hexon with 72% identical and 83% functionally similar residues, followed by bovine AdV3 hexon with 71% identities and 82% functional residue conservation. Phylogenetic analysis suggested that EAdV1 and other AdV do not have an immediate common ancestor. PMID- 9229009 TI - The complete sequence of the genomic RNAs of spinach latent virus. AB - We describe the sequence for the complete genome of spinach latent virus (SpLV). Comparisons of this genome with that of the only other complete genome described for a species within the genus Ilarvirus (citrus leaf rugose virus-CiLRV) indicate that while there are marked differences between the RNA 3 of the two viruses, their respective RNAs 1 and 2 share many similarities. However, the putative 2a protein of SpLV contains a C2H2 type "zinc finger"-like motif located towards the carboxy terminal of the protein which is absent in CiLRV and has not been reported for other members of the family Bromoviridae. A second open reading frame (2b), located at a similar position to that described for the cucumoviruses, occurs in the RNA 2 of both SpLV and CiLRV. The putative coat protein of SpLV is similar to that of citrus variegation virus (CVV) and asparagus virus 2 (AV-2), both members of subgroup 2 of the ilarviruses. We have subsequently demonstrated a serological relationship between SpLV and other viruses in subgroup 2 and suggest that SpLV should be included in this subgroup rather than remain in a separate group (subgroup 6). However, while the putative movement protein of SpLV is remarkably similar to that of AV-2, it shows little relationship with the corresponding protein of CVV and the lack of similarity suggests that a recombination event may have occurred in the past. The relationship between the genera Alfamovirus and Ilarvirus is discussed in the light of the data for the genome of SpLV and recently published information for other members of the genus Ilarvirus. PMID- 9229010 TI - Evidence that virion-associated VP1 of avibirnaviruses contains viral RNA sequences. AB - The VP1 encoded by genomic segment B of birnaviruses is generally known to exist as a genome-linked protein (VPg) and as a "free" polypeptide of 90 kDa in virus particles. The guanylylation activity associated with infectious bursal disease virus (IBDV) was demonstrated by incubating purified virus in presence of [alpha 32P] GTP; optimum activity in the 90 kDa form of VP1 was seen in low salt concentration in the presence of 4 mM magnesium ions over a wide range of incubation temperatures. The IBDV VP1 was shown to lack guanyl transferase activity. Northwestern (RNA-protein) blot analysis of purified virus using a radiolabelled cDNA probe consisting of 3' and 5' ends of genomic segment B indicated that both forms of virion-associated VP1 contained viral RNA sequences of which those linked to VPg corresponded to the two genome segments and those linked to the 90 kDa VP1 were probably a short oligonucleotide of the terminal viral RNA sequences. PMID- 9229011 TI - Non-random deletions in human foamy virus long terminal repeat during viral infection. AB - We have characterized a new form of human foamy virus (HFV) non-random deleted long terminal repeat (LTR) sizing 1078-bp, deleted in its U3 region, sensitive to the viral transactivator and functional in an infectious proviral clone. Besides two known HFV LTRs of 1260-bp and 1123-bp, this LTR represents the smallest, designed S. Analysis of the LTR sequence shows the presence of short direct repeats surrounding the deletions, suggesting a mechanism generating deletion by misalignment of the growing strand during replication. Our data suggest that the deleted LTRs, preferentially associated with chronic viral infection, could be related with viral persistence. PMID- 9229012 TI - Heparin-dependent attachment of respiratory syncytial virus (RSV) to host cells. AB - In this study we could demonstrate that heparin (ED50 = 0.32 +/- 0.12 microgram/ml), but not heparan sulphate or chondroitin sulphate C is able to inhibit in vitro infection of cells by respiratory syncytial virus (RSV). In addition, this protective effect of heparin could only be observed, when heparin was present at the time of inoculation. Enzymatic digestion of cell surface glycosaminoglycans with heparinase and heparitinase, but not chondroitin sulphate ABC lyase reduced the effectiveness of RSV-infection. Affinity chromatography experiments, using immobilised heparin further demonstrated that RSV attachment protein G was able to bind specifically to heparin. Therefore heparin-like proteoglycans showed properties required for attachment of RSV to host cells. PMID- 9229013 TI - Bovine rotavirus strain 678 possesses VP4 of serotype P7[5] specificity. AB - Bovine rotavirus strain 678 is the first G8 strain of bovine origin but the literature is confusing as to its P type. In this study, two-way cross neutralization between 678 and 0510, a prototype G6P7[5] virus, was shown by plaque-reduction neutralization assays, establishing the P type of 678 as being P7[5]. The P7[5] specificity of 678 VP4 was reinforced by the finding that the VP8* portion of 678 VP4 had the highest amino acid identity with those of P7[5] bovine rotaviruses. Apparent contradiction with previous serological studies relates to intricacy of antigenicity and immunogenicity of UK VP4 in reassortants. PMID- 9229014 TI - Enhanced core protein production by hepatitis B virus bearing a mutation in the precore region. AB - An HBV DNA tandem dimer bearing GTG instead of the precore initiation codon (p2WPC-), that bearing an amber mutation at precore codon 28 (p2WPCTer) and that of a wild-type were introduced into HepG2 cells. p2WPC- produced no HBeAg, the same amount of HBsAg as the wild-type, and 2-4 times as much HBcAg and progeny virus DNA. p2WPCTer produced no HBeAg, the same amount of HBsAg, 2 times as much HBcAg and a slightly increased amount of progeny virus. The amounts of p21c peptide in both mutants determined by immunoblotting correlated well with the ELISA titers. These results suggest that these precore single point mutations are responsible for the enhanced core peptide production. PMID- 9229015 TI - RNAs 4A and 5 are present in tomato aspermy virus and both subgroups of cucumber mosaic virus. AB - Primer extension analysis was used to determine the presence of RNAs 4A and 5 in tomato aspermy virus (TAV) and both subgroups of cucumber mosaic virus (CMV). RNAs 4A and 5 were detected in TAV and all CMV strains (representative of both subgroups) that were analysed, except for one subgroup I CMV strain which lacked detectable RNA 5. In subgroup II CMV strains the RNA 5 population was found to consist of two sequence variants. Comparison of the RNA 5 sequences from TAV and CMV indicated that TAV and subgroup II CMV RNA 5 share a much greater degree of sequence similarity than either has with subgroup I RNA 5. RNA 4A and the encoded 2b protein appear to be unique to the cucumovirus genus of the tripartite viruses, which share an otherwise common genome structure, and may have played a role in the evolutionary origin of this genus. PMID- 9229016 TI - Proposals for changes in luteovirus taxonomy and nomenclature. PMID- 9229017 TI - Is there a place for traditional midwives in the provision of community-health services? AB - Traditional midwives (TM) have been involved in delivering babies, and providing a broad range of other services to women, for hundreds of years. They are usually local women with little formal education. As they are well known in their communities they are often called to assist women at the time of delivery. Two opposite views persist about the continuation of their role; some health workers would like to see them trained better and incorporated into the formal health system. Other health workers feel that all deliveries should be attended by either a nurse/midwife or a doctor, and that TM should eventually be phased out. Traditional midwives currently perform > 60% of deliveries in some developing countries and have had their role expanded in some places. Recruiting nurse/midwives and doctors to work in remote areas remains difficult and, even if they were recruited, there is no guarantee that their obstetric services would be used. The evaluation of training programmes has not produced any clear-cut answers in the debate about the long-term role or existence of TM. Rather, the studies have shown that the success of the programmes depends on the resources available, the people involved in the training and how the training is carried out. Some of the lessons learnt from working with TM apply to any two groups of people working together. If TM are going to be offered training, and this must be a local decision made after consultation and an evaluation of prevailing resources and conditions, the training should be a two-way process, with both parties learning from each other. PMID- 9229018 TI - The activity of triple combinations of antifolate biguanides, with and without folinic acid, against plasmodium falciparum in vitro. AB - At least two triple combinations of biguanides, proguanil-atovaquone-dapsone and PS-15-atovaquone-dapsone, may be useful in treating drug-resistant infections of falciparum malaria. Each of these triple combinations can be considered as two synergistic combinations: proguanil-atovaquone and cycloguanil-dapsone, and PS-15 atovaquone and WR99210-dapsone, respectively. Since folinic acid might mitigate the possible side-effects produced by such drug combinations, both combinations were administered, with and without folinic acid, to 16 Saimiri sciureus monkeys. The antimalarial activity in serum samples collected from each monkey, 3, 6 and 24 h after drug administration, was then determined in vitro, against Plasmodium falciparum. The addition of folinic acid had no apparent affect on the antimalarial activity of the triple combinations tested. Such combinations may be useful against Pneumocystis carinii and Toxoplasma gondii in immunocompromised patients, as well as against malaria. PMID- 9229019 TI - Synthetic polypeptides corresponding to the non-repeat regions from the circumsporozoite protein of Plasmodium falciparum: recognition by human T-cells and immunogenicity in owl monkeys. AB - Synthetic polypeptides encompassing the non-repeated regions of the circumsporozoite protein (CSP) of Plasmodium falciparum are very immunogenic in mice and are recognized by sera from donors living in regions where malaria is endemic, both in Africa and South America. Long polypeptides, encompassing the N- or C-terminal regions, have now been used to demonstrate peptide-specific T cells in donors living in an endemic area of Colombia. Although the N-terminal peptide (22-125) was recognized almost exclusively by donors from the endemic area, the patterns of recognition of the C-terminal peptide (289-390) in donors from endemic and non-endemic areas were similar and like the pattern with smaller peptides. The availability of the long polypeptides made it possible to compare T cell responses to the non-repeated regions of the CSP with the presence of peptide-specific antibodies. No correlation was found and no antibodies were detected in donors from non-endemic regions. The long polypeptides also elicited strong antibody and T-cell responses in owl monkeys (Aotus lemurinus). The antibodies generated against the synthetic peptides in such monkeys also recognized sporozoites, the natural infective form of the parasite. The results emphasise the potential of the peptides tested as malaria-vaccine candidates. Not only are they recognized by humans at both the B- and T-cell level but they also elicit strong responses in monkeys and encompass several distinct T-cell epitopes, thus overcoming the limitations of specific, major-histocompatibility complex restriction. PMID- 9229020 TI - Development of different Leishmania major strains in the vector sandflies Phlebotomus papatasi and P. duboscqi. AB - Five lines of four Leishmania major strains, which differ in geographical origin and virulence for mice, were used for experimental infections of Phlebotomus papatasi and P. duboscqi. Differences between the lines, which became evident 6 and 9 days after the infective feed, were more pronounced in P. papatasi. The highest infection rates were found for the more virulent line of strain LV561, while the lowest rates were recorded for strains L119 (low-virulence for mice) and Neal (avirulent for mice). Infection rates depended significantly on the Leishmania strain/line, but not on the vector species. Anterior migration and colonization of the stomodeal valve were observed in flies infected with LV561 and FV1 but infections with other strains were restricted to the whole midgut (L119) or to the abdominal midgut only (Neal). The proportions of the different morphological forms of Leishmania seen in gut smears of infected flies varied considerably with the parasite strain/line. In general, vector forms of LV561 and FV1 were characterized by relatively long flagella and bodies. The strains developing less successfully in vectors tended to have a relatively broad body (L119) or short flagellum (Neal). Transmission experiments were successful with P. duboscqi females infected with the virulent line of LV561. PMID- 9229021 TI - The safety and efficacy of amocarzine in African onchocerciasis and the influence of ivermectin on the clinical and parasitological response to treatment. AB - The hundred men from a forest area of Ghana, without vector control or ivermectin distribution, were randomized to receive a single dose of ivermectin (150 micrograms/kg body weight) on day 1 followed by amocarzine (3 mg/kg twice daily after meals) on days 8, 9 and 10 (34 patients), the ivermectin alone (33 patients) or the amocarzine alone (33 patients). Detailed clinical and laboratory examinations were made before, during and after drug administration. On day 120, all palpable nodules were excised, fixed, sectioned, stained and examined by two blinded observers and the results compared with those for nodules from untreated controls. Mazzotti-type reactions, such as itching, rash, peripheral sensory phenomena and swellings, were more severe or frequent with amocarzine than ivermectin. Pretreatment with ivermectin markedly suppressed these reactions to amocarzine but did not affect other manifestations such as dizziness and gaze evoked nystagmus. Ocular effects were minor in all groups. Ivermectin produced minor macrofilaricidal effects on the adult male worms, marked degeneration of intra-uterine embryos, and potent microfilaricidal effects and suppressed skin microfilariae. Amocarzine did not affect the male worms or the intra-uterine embryos, was a less potent microfilaricide and did not suppress skin microfilariae. The efficacy of ivermectin plus amocarzine was similar to that of ivermectin alone. The present results do not support the findings from the Americas and show that amocarzine has no role in the treatment of onchocerciasis in Africa. PMID- 9229022 TI - Evaluation of invermectin distribution in Benin, Cote d'Ivoire, Ghana and Togo: estimation of coverage of treatment and operational aspects of the distribution system. AB - Invermectin distribution by the Onchocerciasis Control Programme (OCP) was assessed in Benin, Cote d'Ivoire, Ghana and Togo, in terms of the proportion of villages which had been treated and the proportion of villagers in each village treated in the last round who had actually received treatment. These proportions were evaluated both for treatment in the last round of ivermectin distribution and for treatment since the beginning of the drug's distribution in each country. During the last treatment round, 97 (74.6%) of the 130 selected villages investigated in the four countries had received ivermectin treatment, and 67.2% of the members of these 97 treated communities had taken ivermectin. In general, higher percentages of the members of treated villages in Cote d'Ivoire and Ghana had been treated [with mean (S.D.) percentage values of 72.0 (5.2) and 71.6 (4.6), respectively] than in those of Togo [61.8 (5.6)] or Benin [64.2 (4.6)]. Overall, 893 (26.1%) of those interviewed had never received treatment since the beginning of ivermectin distribution but 29.4% had received all the annual treatments. The main reason for non-treatment during the last treatment round was absence from village (54.5% of those not treated), followed by non-eligibility (i.e. pregnant women and young children; 12.2%), refusal to take treatment (2.6%), and shortage of drugs (1.9%). Community approval for the programme was demonstrated when all treated individuals, including those who were absent at the last treatment round, said they would take the ivermectin during the next treatment. During the last treatment round, members of the community assisted in the distribution of the ivermectin tablets in 69 (71.1%) of the 97 treated villages which were investigated. Although only 26 (26.8%) of these 97 villages preferred community-based distribution of ivermectin to the 'mobile' method, it is believed that, with good education and efficient organization, the communities could be encouraged to undertake community distribution. PMID- 9229023 TI - Phlebotomine sandflies in a focus of visceral leishmaniasis in a border area of eastern Sudan. AB - A field study was carried out in eastern Sudan, near the Ethiopian border, to investigate the abundance, seasonality, man-biting behaviour and resting sites of sandflies in two areas where visceral leishmaniasis (kala-azar) is endemic: Umsalala village, Galabat Province; and the adjacent Dinder National Park (DNP), Dinder Province. Abundance of the different species was determined from collections made, using light and sticky-paper traps, in various habitats between November 1993 and February 1995. Man-biting sandflies were collected as they landed on human bait. The habitats investigated for day-resting sandflies were thatched huts, chicken coops, tree-holes, termite mounds and soil cracks. Animal burrows were not investigated. The species found were Phlebotomus (Larroussius) orientalis, P. (Phlebotomus) papatasi, P. (Paraphlebotomus) saevus, P. (Anaphlebotomus) rodhaini, Sergentomyia (Sintonius) clydei, S. (Sergentomyia) antennata, S. (Sergentomyia) sckwetzi, S. (Parrotomyia) africana and S. (Grassomyia) squamipleuris. Phlebotomus orientalis was the only man-biting sandfly species found in the DNP whereas P. papatasi, P. orientalis and P. saevus were all found in Umsalala. Abundance of each species varied with the habitat. In Umsalala and a camp for game wardens in the DNP, Sergentomyia spp. predominated over Phlebotomus. In the DNP, the most abundant sandfly in a thicket dominated by Acacia seyal trees was P. orientalis, followed by Sergentomyia spp. Significant habitat 'preferences' were observed for most sandfly species in the area. In attempts to find resting flies, P. orientalis was only found resting in the mounds made by the termite Macrotermes herus and P. papatasi was only found inside huts; no resting sites were detected for other Phlebotomus spp. but Sergentomyia spp. were observed in all the sites investigated. The P. orientalis in the DNP showed a clear seasonal variation in abundance, which was closely correlated with the mean monthly temperature and relative humidity of the area. A remarkable increase in the abundance of this vector occurred at the beginning of the rainy season. PMID- 9229024 TI - Use of discontinuous Percoll gradients to isolate Cyclospora oocysts. PMID- 9229025 TI - Infections with hepatitis B virus in three villages endemic for schistosomiasis japonica in the Dongting Lake region of China. PMID- 9229026 TI - Clinical laboratory techniques in a developing country (Burundi). PMID- 9229027 TI - Suicide as an outcome for mental disorders. A meta-analysis. AB - BACKGROUND: Mental disorders have a strong association with suicide. This meta analysis, or statistical overview, of the literature gives an estimate of the suicide risk of the common mental disorders. METHOD: We searched the medical literature to find reports on the mortality of mental disorders. English language reports were located on MEDLINE (1966-1993) with the search terms mental disorders', 'brain injury', 'eating disorders', 'epilepsy', 'suicide attempt', 'psychosurgery', with 'mortality' and 'follow-up studies', and from the reference lists of these reports. We abstracted 249 reports with two years or more follow up and less than 10% loss of subjects, and compared observed numbers of suicides with those expected. A standardised mortality ratio (SMR) was calculated for each disorder. RESULTS: Of 44 disorders considered, 36 have a significantly raised SMR for suicide, five have a raised SMR which fails to reach significance, one SMR is not raised and for two entries the SMR could not be calculated. CONCLUSIONS: If these results can be generalised then virtually all mental disorders have an increased risk of suicide excepting mental retardation and dementia. The suicide risk is highest for functional and lowest for organic disorders with substance misuse disorders lying between. However, within these broad groupings the suicide risk varies widely. PMID- 9229028 TI - Outcome in schizophrenia and related disorders compared between developing and developed countries. A recursive partitioning re-analysis of the WHO DOSMD data. AB - BACKGROUND: Data on the two-year pattern of course of illness have been collected in the WHO study of the Determinants of Outcomes of Severe Mental Disorder (DOSMD). These data are reanalysed using recursive partitioning, a method not yet applied to psychiatric data to test the hypothesis that subjects from participating centres in developing countries had better outcomes than those in developed countries. METHOD: Subjects were those from the DOSMD study for whom two-year follow-up data were available (n = 1056). The classification and regression trees recursive partitioning technique was used to examine the predictor variables associated with the outcome variable two year pattern of course. RESULTS: Pattern of course was best predicted by centre, but two developed centres (Prague and Nottingham) grouped with the developing country centres excluding Cali, having better outcomes than in the remaining developed country centres and Cali. Type of onset (insidious v. non-insidious) was the next strongest predictor, but its effect differed across these two centre groupings. Effects for some groups were modified by other predictor variables, including age, child and/or adolescent problems, and gender. CONCLUSIONS: The predominant predictor effects on two-year pattern of course continued to be centre and type of onset, but complex interactions between these variables and other predictor variables are seen in specific centre groupings not strictly defined by 'developing' and 'developed'. PMID- 9229029 TI - Is the earlier age at onset of schizophrenia in males a confounded finding? Results from a cross-cultural investigation. AB - BACKGROUND: The finding of an earlier age at onset of schizophrenia in males compared with females, replicated across a number of studies, appears to be so robust as to support hypotheses about gender differences in the aetiology of the disorder. However, the possibility that this observed gender effect might reflect other confounding variables has not been adequately explored. METHOD: We analysed data on 778 men and 653 women, in three developing countries and in seven developed countries, who had been assessed in the WHO 10-country study of schizophrenia. We applied a generalised linear modelling strategy to estimate the unconfounded contributions of gender, family history, premorbid personality and marital status to age at onset. RESULTS: The model that explained the highest percentage of the total variance indicated strong main effects (P < 0.001) for marital status and premorbid personality, a weak effect for family history, and an attenuated effect for gender. Two independent verification procedures suggested an independent onset-delaying effect for marital status (married), more marked in males. CONCLUSIONS: The gender difference in the age at onset of schizophrenia is not a robust biological characteristic of the disorder. Failure to control for marital status and premorbid personality in male/ female comparisons of age at onset may explain a large part of the differences reported previously. PMID- 9229030 TI - Psychopathological syndromes and familial morbid risk of psychosis. AB - BACKGROUND: Familial liability in the functional psychoses had traditionally been examined by comparing mutually exclusive diagnostic categories. This study examines overlapping psychopathological dimensions in relation to familial morbid risk of psychosis. METHOD: We tested for associations between seven factor analysis derived psychopathological dimensions and familial morbid risk of psychosis, in a sample of 150 patients with recent-onset functional psychosis and 548 of their first-degree relatives. RESULTS: A syndrome characterised by affective blunting and insidious and early onset of illness, non-specifically predicted psychosis in the first-degree relatives, whereas a manic syndrome specifically predicted affective psychosis in the relatives. No other main effects were observed, but there were interactions with proband diagnosis: a syndrome characterised by bizarre behaviour, inappropriate affect, catatonia and poor rapport predicted psychosis in relatives of schizophrenic probands, and a syndrome of depressive: symptoms predicted psychosis in relatives of schizoaffective probands. Positive symptoms were not associated with illness in the relatives. CONCLUSIONS: Genetic effects in the functional psychoses may comprise non-specific components that canalise a general, early-onset, affective blunting phenotype and several other, more specific, influences on phenotypic variation. PMID- 9229031 TI - The Nottingham Acute Bed Study: alternatives to acute psychiatric care. AB - BACKGROUND: Although modern psychiatric services seek alternatives to hospitalisation wherever appropriate, the national trend toward higher bed occupancies on acute psychiatric wards has refocused attention on community-based alternatives and methods of assessing reed for acute care. METHOD: We surveyed key decision makers in a community-oriented district service with a low acute psychiatric bed to population ratio, in order to examine alternatives to hospitalisation in a cohort of consecutive admissions over a six-month period. RESULTS: Alternatives to acute ward hospitalisation were identified for 29% of admissions, and for 42% of those with an admission duration of more than 60 days. Residential options were chosen more often than intensive community support. Simulated bed day savings were considerable. CONCLUSIONS: In a community-oriented service, key decision-makers could identify further alternatives to acute ward hospitalisation, although relatively few non-residential, community support options were chosen. Although this methodology has limitations, data based upon keyworker judgements probably have greater local 'ownership', and the option appraisal process itself may challenge stereotyped patterns of resource use. PMID- 9229032 TI - Mental health review tribunals. A follow-up of reviewed patients. AB - BACKGROUND: Mental health legislation allows for treatment needs to override civil liberty. Mental health review tribunals act as a counterbalance. This study examines the long-term outcome of patients reviewed by a tribunal, and in particular whether the tribunal, in its concern for civil liberty, might be discharging patients prematurely. METHOD: All non-offender patients from a defined catchment area reviewed by the tribunal between the inception of the 1983 Mental Health Act and 31 December 1991 were followed-up until 31 May 1993. RESULTS: Those discharged by the tribunal did not differ significantly from those refused discharge in subsequent survival period in the community, in readmission rate or in final outcome. CONCLUSIONS: Within the limitations of a non experimental study, the main hypothesis was not supported. An intensive study of family and personal life in the three months after discharge would cast useful additional light on the soundness of tribunal decisions. PMID- 9229033 TI - Cost-effectiveness of antidepressant treatment reassessed. AB - BACKGROUND: A recent simulation concluded that the serotonin-specific reuptake inhibitor (SSRI) paroxetine was more cost-effective than the tricyclic antidepressant (TCA) imipramine, despite substantially higher medication acquisition costs. METHOD: We replicated the previous model and revised key assumptions which drove the results. The revised model was subjected to sensitivity analysis. RESULTS: Most scenarios in the revised model showed that the TCA is equally or more cost-effective than the SSRI. Model revision producing these results were changes in assumptions about switched treatment success rates, treatment length and initial treatment success. The revised model appears sensitive to drug acquisition and delivery costs and costs of treatment failure. CONCLUSIONS: Based on the model, a policy of using TCAs as first-choice antidepressant treatment, with SSRIs reserved for those patients not doing well initially, appears more cost-effective than the reverse sequence. Given limitations in current knowledge about key parameters to include in a simulation model, large prospective random-assignment cost-effectiveness studies are needed. PMID- 9229034 TI - Generalisability of results from randomised drug trials. A trial on antimanic treatment. AB - BACKGROUND: Exemplified by a randomised trial on antimanic treatment, this paper addresses the question of whether selection of patients for drug trials may limit the applicability of study results from the randomised patients to a wider population. METHOD: During two-year period, all consecutively admitted patients from a defined catchment area were screened for inclusion criteria concerning age, diagnosis and severity of illness. The subsequently excluded subgroups of patients were compared with the randomised patients by multivariate data analysis. RESULTS: One hundred and sixty-four patients met the inclusion criteria. However, after exclusion for various reasons, only 27 (17%) patients remained for randomisation. The randomised patients and the excluded patients differed substantially. CONCLUSIONS: The generalisability of trial results is limited. Reports of randomised drug trials should carefully describe the screening procedure for inclusion and, when possible, present relevant comparisons-between the randomised patients and the various subgroups of excluded patients. PMID- 9229035 TI - The effect of treatment on the two-year course of late-life depression. AB - BACKGROUND: The purpose of this study was to examine the effect of treatment on the long-term course of geriatric depression. METHOD: Eighty-four elderly patients who had responded to treatment of the index episode of major depression were maintained on full-dose antidepressant medication and followed on a monthly basis for two years. Relapse and recurrence were treated in a systematic manner. RESULTS: The cumulative probability of surviving for two years without relapse or recurrence was 74%. Of the 14 patients who suffered recurrence following recovery from the index episode, all responded to a change of treatment, and 71% remained well for the remainder of the study. The risk of recurrence was significantly increased by a delayed response to treatment of the index episode. CONCLUSIONS: Continuation and maintenance treatment with full-dose antidepressant medication, frequent follow-up, and vigorous treatment of relapses and recurrences, were associated with a good outcome in this group of elderly patients. PMID- 9229036 TI - The use of seclusion and emergency medication in a hospital for people with learning disability. AB - BACKGROUND: The management of disturbed behaviour in facilities for those with learning disabilities involves a spectrum of approaches including the prescription of emergency medication, restraint and seclusion. The use of these techniques has recently come under close scrutiny. METHOD: All incidents requiring emergency medication or seclusion that occurred in a large hospital for those with learning disabilities were studied over a six-months period. The precipitating factors, course and outcome of those who had received emergency medication or seclusion were then examined. RESULTS: In all, 286 incidents involving 72 individuals occurred during the study period. The episodes requiring seclusion comprised 19% of all incidents. Two-thirds of the patients involved were male but six female patients accounted for 36% of all incidents. During the second part of the study, when the staff knew that the treatments used were being monitored, there was a significant reduction in use of restraint and emergency drugs given intramuscularly. Patients receiving seclusion were judged to have a better outcome one hour after the onset of the incident compared with those who received medication. CONCLUSIONS: Despite concerns about the use of seclusion, the results of this survey suggest that procedures that remove the patient from the environment contributing to the disturbance may have certain advantages in this population. PMID- 9229037 TI - Cytogenetic abnormalities on chromosome 18 associated with bipolar affective disorder or schizophrenia. AB - BACKGROUND: A few recent linkage studies have shown a possible locus for bipolar disorder on chromosome 18. Cytogenetic studies may assist in the further localisation of susceptibility loci on this chromosome. METHOD: A search was made for abnormalities of chromosome 18 in two separate large cytogenetic databases. In Denmark detection of mental illness in subjects with chromosome abnormalities was done by cross-linking the two separate register of psychiatric and chromosome disorders. In Scotland the Cytogenetic Registry of the MRC Human Genetics Unit undertakes long-term clinical follow-up of all cases with chromosome abnormalities. RESULTS: Cross-linking the two Danish register's revealed a family with the rare karyotype abnormality inv(18) (p11.3;q21.1) with one inversion carrier who also suffered from bipolar disorder. In this family there were two other cases of bipolar disorder, but the karyotype of these cases could not be established. One family in Scotland showed a case of schizophrenia in a carrier of inv(18) with the same breakpoints as the Danish family. CONCLUSIONS: We suggest further studies of the 18p11.3 and 18q21.1 regions in order to identify genes involved in bipolar affective disorder and schizophrenia. PMID- 9229038 TI - A transcultural pattern of drug use: qat (khat) in the UK. AB - BACKGROUND: This study investigates patterns of qat use among 207 Somalis living in London. METHOD: Subjects were recruited using privileged access interviewing. Somalian interviewers were recruited who shared the same culture as the subjects. Data were collected by means of a structured interview. RESULTS: One hundred and sixty-two subjects (78%) had used qat. The majority (76%) used more qat than in Somalia. Some users reported moderate dependence; a minority reported severe problems. Adverse psychological effects included sleep problems, anxiety and depression. Medical problems associated with qat use were rare. CONCLUSIONS: Qat users who continue to use this drug when it is transplanted from a traditional context may experience difficulties. Qat use can also be seen as playing a positive role in supporting the cultural identity of the Somalian community. Severe problems were rarely reported. Qat consumption should be considered when addressing health-related topics with patients from those communities in which qat use is common. PMID- 9229039 TI - Maintenance electroconvulsive therapy and cognitive function. AB - BACKGROUND: ECT is rarely used as a prophylactic treatment. A 74-year-old woman with unstable bipolar affective disorder receiving maintenance ECT presented a unique opportunity to measure the cognitive effects of continuing ECT. METHOD: A single case report with serial psychometric testing during over 400 ECT treatments as a single maintenance treatment. RESULTS: Serial testing did not demonstrate progressive cognitive deterioration, but consistent cognitive deficits typical of acute treatment were evident. The degree of cognitive difficulty may be related to the frequency of treatment. CONCLUSIONS: Maintenance ECT can be an effective prophylactic treatment for selected patients. Cognitive effects would appear to be no greater than with acute treatment and seem to be non-progressive. PMID- 9229040 TI - Antidepressant withdrawal syndrome. PMID- 9229041 TI - Recent registration and referrals from general practitioners. PMID- 9229042 TI - Lipid-lowering drugs and mortality. PMID- 9229043 TI - Perpetrators of child sexual abuse. PMID- 9229044 TI - Urine screening for drugs. PMID- 9229045 TI - Acute clozapine overdosage. PMID- 9229046 TI - Air pollution and other local factors in respiratory disease. 1959. PMID- 9229048 TI - That which we call social medicine.... PMID- 9229049 TI - Has the prevalence of asthma increased in children? Evidence from a long term study in Israel. AB - BACKGROUND: The permit to build and operate the first 1400 megawatt coal fired power plant in Israel was given provided that three monitoring systems environmental, agricultural, and health monitoring-be set up near the plant. This study was carried out in the framework of a health monitoring system which included a mortality survey, requests for health services, a schoolchildren's health survey, and an adult panel study. METHODS: 2nd, 5th, and 8th grade school children living in three communities with different expected levels of air pollution were followed up every three years. They performed pulmonary function tests (PFT), and their parents filled out American Thoracic Society-National Heart and Lung Institute (ATS-NHLI) health questionnaires. A follow up of the prevalence of respiratory conditions among the studied schoolchildren in four rounds of tests was carried out. This report deals with the changes in the prevalence of asthma, related respiratory conditions, and PFT in the data sets gathered among 5th grade schoolchildren. RESULTS: A significant (p = 0.0024) increase in the prevalence of asthma could be observed among 5th grade children in all three communities studied between 1980 and 1989. At the same time a significant (p = 0.0172) rise in the prevalence of wheezing accompanied by shortness of breath could be observed. A similar trend could not be found for the prevalence of bronchitis and other respiratory conditions among the studied children. PFT (FEV1, FEV1/FVC) of children suffering from asthma or from wheeze accompanied by shortness of breath were lower than those of healthy children. Changes in the prevalence of background variables over time could not explain the significant rise in the prevalence of asthma among the children. CONCLUSIONS: The significant rise in asthma and related respiratory conditions coupled with reduced PFT observed in this study suggest that the increase over time in the prevalence of asthma is a true increase in morbidity and not due to reporting bias. The increased prevalence of asthma could be observed in all the communities studied and does not seem to be connected with the operation of the power plant. PMID- 9229050 TI - Asthma and thunderstorms: description of an epidemic in general practice in Britain using data from a doctors' deputising service in the UK. AB - OBJECTIVE: To describe the areas affected and the scale of an epidemic of thunderstorm associated asthma on the night of 24/25 June 1994 and to explore the spatial and temporal relationship between the thunderstorm and the associated epidemic. SETTING: The 29 offices of a deputising service for general practitioners' (GP) out of hours calls (Healthcall). At the time of the storm the deputising service provided out of hours cover for about 8500 out of about 30000 GPs in England, Scotland, and Wales. METHODS: Patients who phoned the Healthcall offices to request a home visit were categorised as "asthma" or "other causes" based on their presenting complaint. The number of calls on the night of 24/25 June 1994 was compared in areas affected by thunderstorms and areas not affected by thunderstorms and with the night of 17/18 June 1994, when there were no thunderstorms. RESULTS: A large area of the south and east of England was affected by an epidemic of asthma closely related both temporally and spatially with the thunderstorms on 24/25 June 1994. The pooled Mantel-Haenszel estimate for the risk of asthma in thunderstorm affected areas compared with the control night was 6.36 (95% confidence interval 4.97, 8.32) compared with a value of 1.01 (0.80, 1.27) for those not exposed. Extrapolation suggests about 1500 extra patients were likely to have requested a visit from a GP that night because of epidemic asthma. CONCLUSIONS: Under certain circumstances thunderstorms are associated with asthma and can affect many patients. Deputising services are a useful source of data for the investigation of epidemics in primary care. PMID- 9229051 TI - A comprehensive study of smoking in primary school children in Hong Kong: implications for prevention. AB - STUDY OBJECTIVE: To identify factors associated with smoking behaviour in primary school children in Hong Kong. DESIGN: A cross sectional survey in which both children and parents completed questionnaires. The main outcome measure was the smoking status of the children; and risk factors (knowledge of and attitude to smoking and demographic and socioeconomic background) were identified as predictors of ever/never smoking. SETTING AND SUBJECTS: Altogether 9598 primary school children, aged 8-13 years, and attending 27 schools from two districts in Hong Kong participated. MAIN RESULTS: The prevalence of ever-smoking was 12% (1119)-15% (760) in boys and 7% (359) in girls. It ranged from 3% in 8 year old girls to 52% in 13 year old boys. The factors associated with ever-smoking included the following: being a boy (adjusted odds ratio 2.21; 95% confidence interval 1.89, 2.59), increasing age per year (1.48; 1.40, 1.57), living in Kwai Tsing district (1.29; 1.10, 1.50), having one or more smokers at home (2.07; 1.78, 2.39), and having a father who was not working (1.41; 1.19, 1.67). Children who were ever-smokers had both seen and approved of their friends' smoking (8.79; 5.33, 14.50), had a more positive attitude towards smoking (3.35; 2.21, 5.09), and were more successful in recognising cigarette brand names and logos (1.67; 1.42, 1.96), but they lacked confidence (1.78; 1.32, 2.39). CONCLUSIONS: The influences on child smoking are multifactorial and programmes in Hong Kong are falling to curb them. The control of these risk factors must be incorporated in the smoking prevention policy of Hong Kong and supported by future enforced legislation. PMID- 9229052 TI - Smoking during pregnancy: how reliable are maternal self reports in New Zealand? AB - OBJECTIVE: To determine the reliability of self reports of smoking during pregnancy. METHODS: Residual sera from early and late antenatal blood samples were tested for cotinine for all pregnancies over a six month period. Over an overlapping 12 month period, a postal questionnaire on smoking was also sent to all new mothers (n = 4857) when their baby was 4-8 weeks old. Smoking status from obstetric booking notes was also obtained. RESULTS: The cotinine-validated smoking prevalence was 31.3% for the first trimester and 27.7% for the third trimester. Questionnaire self reported prevalences were 19.2% and 15.7% for the first and third trimesters respectively, and 18.9% for obstetric booking. Of cotinine-validated smokers, 22% denied smoking-self deceivers. Of mothers who replied to the questionnaire, a half appeared to systematically under report the amount they smoked. CONCLUSIONS: Nearly a quarter of smoking pregnant women did not report smoking. Moreover, of those who did, the amount smoked was often under reported. This tendency to under report may rise as pressures to stop smoking increase. Accurate measures of smoking prevalence in pregnant women will require objective testing. PMID- 9229053 TI - Smoking and relative body weight: an international perspective from the WHO MONICA Project. AB - STUDY OBJECTIVE: To investigate the magnitude and consistency of the associations between smoking and body mass index (BMI) in different populations. DESIGN: A cross sectional study. SETTING AND PARTICIPANTS: About 69,000 men and women aged 35-64 years from 42 populations participating in the first WHO MONICA survey in the early and mid 1980s. MAIN RESULTS: Compared to never smokers, regular smokers had significantly (p < 0.05) lower median BMI in 20 (men) and 30 (women) out of 42 populations (range -2.9 to 0.5 kg/m2). There was no population in which smokers had a significantly higher BMI than never smokers. Among men, the association between leanness and smoking was less apparent in populations with relatively low proportions of regular smokers and high proportions of ex-smokers. Ex-smokers had significantly higher BMI than never smokers in 10 of the male populations but in women no consistent pattern was observed. Adjustment for socioeconomic status did not affect these results. CONCLUSIONS: Although in most populations the association between smoking and BMI is similar, the magnitude of this association may be affected by the proportions of smokers and ex-smokers in these populations. PMID- 9229054 TI - Weight gain during the first year of life in relation to maternal smoking and breast feeding in Norway. AB - OBJECTIVE: To assess the weight gain during the first year of life in relation to maternal smoking during pregnancy and the duration of breastfeeding. DESIGN: This was a one year cohort study. SETTING: The city of Oslo, Norway. PARTICIPANTS: Altogether 3020 children born in Oslo in 1992-93. Children were divided into three groups as follows: 2208 born to non-smoking mothers, 451 to mothers who were light smokers (< 10 cigarettes per day), and 261 to mothers who were heavy smokers (> or = 10 cigarettes per day). MAIN RESULTS: The mean birth weights were 3616 g, 3526 g, and 3382 g and 1 year body weights were 10,056 g (gain 6440 g per year), 10,141 g (6615 g), and 10,158 g (6776 g) in children of non-smoking and light and heavy smoking mothers respectively. Cox regression analysis showed that children of heavy smokers were 2.0 (95% confidence interval, 1.7, 2.3) times and children of light smokers 1.3 (1.2, 1.5) times more likely to have stopped breast feeding during their first year of life compared with children whose mothers were non-smokers. Linear regression analysis, adjusting for confounders, showed that weight gain was slower in breast fed children than in those who were not breast fed (-38 g (-50, -27) per month of breast feeding). Compared with children of non smokers, the adjusted weight gain was 147 g (40, 255) per year greater in children of light smokers and 184 g (44, 324) per year in children of heavy smokers. CONCLUSION: Children catch up any losses in birth weight due to maternal smoking, but some of the catch up effect is caused by a shorter duration of breast feeding in children of smoking mothers. PMID- 9229055 TI - Cardiovascular risk factor profile in subjects with familial predisposition to myocardial infarction in Denmark. AB - STUDY OBJECTIVES: To identify possible modifiable mediators of familial predisposition to myocardial infarction (MI) by assessing the risk factor profile in individuals without MI in relation to parental occurrence of MI. DESIGN AND METHODS: Cross sectional survey of the general population. The odds of an adverse cardiovascular risk factor profile in subjects reporting parental occurrence of MI versus subjects not reporting parental occurrence were estimated by logistic regression models. SETTING: The Copenhagen Centre for Prospective Population Studies, where subjects investigated in three Danish prospective population studies are integrated. PARTICIPANTS: Subjects were 9306 females and 11,091 males aged 20-75 years with no history of MI. A total of 1370 subjects reported maternal MI and 2583 reported paternal MI. MAIN RESULTS: Increased systolic and diastolic blood pressure, increased cholesterol level, low ratio between high density lipoprotein (HDL) and total cholesterol (TC), and heavy smoking, were more frequent in subjects with parental occurrence of MI than in controls irrespective of sex and age of the subjects. Maternal MI was more predictive for increased cholesterol and decreased HDL/ TC ratio than paternal MI, and the risk of an increased cholesterol level was higher in subjects aged 20-39 years than in older subjects. No differences in body mass index, triglycerides, and physical inactivity were observed. CONCLUSIONS: Subjects free of previous MI who reported a parental occurrence of MI had an adverse cardiovascular risk factor profile regarding systolic and diastolic blood pressure, total cholesterol, the ratio between HDL and total cholesterol, and smoking. Thus, these modifiable risk factors may be mediators of the familial predisposition to MI. PMID- 9229057 TI - Nutritional assessment of two famine prone Ethiopian communities. AB - STUDY OBJECTIVES: To compare two ethnically distinct Ethiopian populations (Oromo Arsi in Elka in the Rift Valley and Anyuak in Punjido in Gambella) for two widely used anthropometric indices of protein-energy malnutrition: body mass index < 18.5 and arm muscle circumference < 80% of the median of the US NHANES reference data. DESIGN: Anthropometric measurements were made in two cross sectional community surveys. SETTING: The Elka village in the central Rift Valley and the Punjido village in western Ethiopia. PARTICIPANTS: 1170 and 560 people from all age groups in Elka and Punjido, respectively. MAIN RESULTS: Estimates of the prevalence of malnutrition in each group differed considerably when defined from the body mass index, but were quite similar when the arm muscle circumference was used. Data for children indicated that the boys and girls in one group (Punjido) were taller but had about the same weights for age as those in the other group (Elka), suggesting that the low body mass indices among the Punjido might have a genetic basis. CONCLUSIONS: Body mass index systematically overestimates the prevalence of malnutrition among the Anyuaks in Punjido. Local reference data from a well nourished Anyuak sample or from an ethnically related population is needed to evaluate appropriately malnutrition using the body mass index. This study shows that care must be taken when assessing different ethnic groups using existing international anthropometric references. PMID- 9229056 TI - Political changes and trends in cardiovascular risk factors in the Czech Republic, 1985-92. AB - BACKGROUND: Mortality from cardiovascular diseases is substantially higher in central and eastern Europe than in the west. After the fall of communism, these countries have undergone radical changes in their political, social, and economic environments but little is known about the impact of these changes on health behaviours or risk factors. Data from the Czech Republic, a country whose mortality rates from cardiovascular diseases are among the highest, were analysed in this report. OBJECTIVES: To examine the trends in cardiovascular risk factors in Czech population over the last decade during which a major and sudden change of the political and social system occurred in 1989, and whether the trends differed in relation to age and educational group. DESIGN AND SETTING: Data from three cross sectional surveys conducted in 1985, 1988, and 1992 as a part of the MONICA project were analysed. The surveys examined random samples of men and women aged 25-64 in six Czech districts and measured the following risk factors: smoking, blood pressure, body mass index (BMI), and total and high density lipoprotein (HDL) cholesterol. RESULTS: The numbers of subjects (response rate) examined were 2573 (84%) in 1985, 2769 (87%) in 1988, and 2353 (73%) in 1992. Total cholesterol and body mass index increased between 1985 and 1988 and decreased between 1988 and 1992. The prevalence of smoking was declining slightly in men between 1985 and 1992 but remained stable in women. There were only small changes in blood pressure. The decline in cholesterol and BMI in 1988-92 may be related to changes in foods consumption after the price deregulation in 1991. An improvement in risk profile was more pronounced in younger age groups, and the declines in cholesterol and obesity were substantially larger in men and women with higher education. By contrast, there was an increase in smoking in women educated only to primary level. CONCLUSION: Substantial changes in cholesterol, obesity, and women's smoking occurred in the Czech population after the political changes in 1989. Although a causal association cannot be claimed, national trends in foods consumption are consistent with changes in blood lipids and obesity. Further monitoring of trends is required to confirm these trends. PMID- 9229058 TI - Effect of labour market conditions on reporting of limiting long-term illness and permanent sickness in England and Wales. AB - STUDY OBJECTIVE: To identify any bias in the reporting of limiting long term illness and permanent sickness due to labour market conditions, and show the absence of the effect in mortality rates. DESIGN: A geographically based study using data from the 1991 census. Standardised ratios for mortality and long term illness in people aged 0-64 years and permanent sickness in people of working age were compared with Carstairs deprivation scores in multilevel models which separated the effects operating at three geographical scales: census wards, travel to work areas, and standard regions. Holding ward and regional effects constant, variations between travel to work areas were compared with long term unemployment rates. SETTING: Altogether 8690 wards and 262 travel to work areas in England and Wales. MAIN RESULTS: Variations in mortality, limiting long term illness, and permanent sickness were related to Carstairs deprivation scores and standard region. With these relationships controlled, limiting long term illness and permanent sickness were significantly related to long term unemployment levels in travel to work areas, but mortality was not affected. Self reported morbidity was more sensitive to variations in long term unemployment rates in conditions of high social deprivation than in affluent populations. CONCLUSIONS: Limiting long term illness and permanent sickness measures may reflect a tendency for higher positive response in difficult labour market conditions. For average social deprivation conditions, standardised limiting long term illness for people aged 0-64 years was 20% higher in travel to work areas where employment prospects were relatively poor compared with areas with relatively good employment prospects. This casts doubt on the use of limiting long term illness as an indicator of objective health care needs for resource allocation purposes at national level. PMID- 9229060 TI - Community mothers' programme: extension to the travelling community in Ireland. AB - STUDY OBJECTIVE: To see whether the community mothers' programme, using lay volunteer mothers to deliver a childhood development programme, could be extended successfully to the travelling community in Ireland. DESIGN: This was a prospective study of the travelling community; comparisons were made with results of a previous randomised trial of settled mothers. SETTING: A regional health authority in Ireland. PARTICIPANTS: These comprised 39 traveller and 127 settled intervention mother/ infant pairs (randomised controlled trial (RCT) intervention); settled community mothers; 105 settled control pairs (RCT control). All mothers received standard support; traveller and RCT intervention groups also received the services of a community mother. MAIN RESULTS: The travellers' sociodemographic profile differed significantly from the other groups. At the end of the study, traveller and intervention children were exposed to more cognitive games and nursery rhymes. There were significant differences in the proportions who received all three shots of their primary immunisation schedule before 12 months of age and who received "three in one" vaccination, with traveller children doing least well. The diet of traveller children surpassed that of RCT controls in all food groups except fruit; they were less likely to begin cows' milk before 26 weeks of age. Traveller mothers' diet was superior to that of RCT control and similar to RCT intervention mothers. Traveller and RCT intervention mothers were less likely to feel tired, feel miserable, and want to stay indoors than RCT control mothers. CONCLUSIONS: The results of the community mothers' programme in the travelling community are encouraging; poor immunisation uptake remains a challenge. PMID- 9229059 TI - Estimating potential savings in cancer deaths by eliminating regional and social class variation in cancer survival in the Nordic countries. AB - STUDY OBJECTIVES: To examine equity in the health care system with regard to cancer patient care by estimating the level of systematic regional variation in cancer survival in the Nordic countries. Specifically, those cancer sites which exhibit high levels of systematic regional variation in survival and hence inequity were identified. Estimating the reduction in cancer deaths which could be achieved by eliminating this variation so that everyone receives effective care will provide a readily interpretable measure of the amount of systematic regional variation. A comprehensive analysis of regional variation in survival has not previously been conducted so appropriate statistical methodology must be developed. SETTING AND PARTICIPANTS: All those aged 0-90 years who had been diagnosed with at least one of 12 common malignant neoplasms between 1977 and 1992 in Denmark, Finland, Norway, and Sweden. DESIGN: A separate analysis was conducted for each country. Regression models for the relative survival ratio were used to estimate the relative risk of excess mortality attributable to cancer in each region after correcting for age and sex. An estimate of the amount of systematic regional variation in survival was obtained by subtracting the estimated expected random variation from the observed regional variation. An estimate was then made of the potential reduction in the number of cancer deaths for 2008-12 if regional variation in survival were eliminated so that everyone received the same level of effective care. MAIN RESULTS: Between 2008 and 2012, an estimated 2.5% of deaths from cancers in the 12 sites studied could be prevented by eliminating regional variation in survival. The percentage of potentially avoidable deaths did not depend on country or sex but it did depend on cancer site. There was no relationship between the level of regional variation in a given country and the level of survival. The cancer sites for which the greatest percentage savings could be achieved were melanoma (11%) and cervix uteri (6%). The sites for which the highest number of deaths could be prevented were prostate, colon, melanoma, and breast. CONCLUSIONS: This methodology showed a small amount of systematic regional variation in cancer survival in the Nordic countries. The cancer sites with high levels of regional variation identified are potential targets for cancer control programmes. PMID- 9229061 TI - Strategies for the prevention of psychiatric disorder in primary care in south London. AB - STUDY OBJECTIVE: To compare the potential impact of high risk and population based approaches to the prevention of psychiatric disorder, using a representative sample of general practice attenders as the target population. DESIGN: This was a prospective cohort study. SETTING: A health centre in south London. PARTICIPANTS: Three hundred and seven consecutive attenders aged 16-65, recruited at randomly selected general practice surgeries. MAIN RESULTS: A linear association was found between the number of different types of socioeconomic adversity reported at recruitment (T1) and the prevalence of psychiatric disorder one year later (T2). The population attributable fraction (PAF) for socioeconomic adversity at T1 was 37.4%. In theory, social interventions for high risk individuals at T1 would reduce the prevalence of psychiatric disorder at T2 by 9% at most, compared with a reduction of 18% if just one item of socioeconomic adversity were eliminated among those with any socioeconomic risk factors. CONCLUSIONS: Social interventions targeted at individuals at highest risk of the most common mental disorders are likely to be extremely limited in their capacity to reduce the prevalence of these conditions. A population based risk reduction strategy, modified according to individual risk, represents a potentially feasible and effective alternative. PMID- 9229062 TI - Profiling outpatient workload: practice variations between consultant firms and hospitals in south west England. AB - OBJECTIVES: To describe the variation in outpatient new to old ratios between consultants and between providers for seven high volume specialties (four surgical, three medical). DESIGN: This was a descriptive study at consultant and provider unit level based upon patient administration system data from the South and West Regional Health Authority for the financial year 1992-93. Additional components of variance analysis was used to distinguish individual consultant effects from host institution effects. SETTING: The former South Western Regional Health Authority area from Gloucestershire to Cornwall. SUBJECTS: Altogether 345 consultant firms in seven specialties grouped into 13 provider unit groups. MAIN MEASURES: New to old ratio, omitting elective inpatients followed up as outpatients. RESULTS: Variation between consultants is greater in surgical than in medical specialties, while absolute levels of new to old ratios tend to be higher in surgical specialties than in medical. Variation between provider unit groups is also greater in surgical specialties. Analysis of variance shows that more total variance is attributable to provider unit group in gynaecology than in other specialties. CONCLUSIONS: Within individual specialties there is evidence of substantial variation that is not attributable to underlying differences in morbidity patterns. There is evidence of marked variation in terms of both individual consultants and institutions, a finding that provides the springboard for further analytical work. Published routine outpatient activity statistics should distinguish between new referrals, inpatient follow up, and clinic rebookings. PMID- 9229063 TI - Emergency admission of patients to general surgical beds: attitudes of general practitioners, surgical trainees, and consultants in Liverpool, UK. AB - OBJECTIVES: To determine (a) whether doctors involved in the process of emergency surgical admission could agree about which patients should be admitted, (b) whether there were consistent differences between doctors in different specialty groups, and (c) whether these opinions were greatly influenced by non-clinical factors. DESIGN: Independent assessment of summarised case histories by three "expert" clinicians (two consultant surgeons and one general practitioner (GP)), by a group of 10 GPs, and by a group of 10 junior and senior surgeons. Experts, but not other observers, scored admissions both independently and as a consensus group. Observers indicated for each patient whether they would admit, would not admit, or were unsure. SETTING: An urban general hospital with teaching status. SUBJECTS: Fifty consecutive patients admitted to the general surgical unit as emergencies during 1995. MAIN OUTCOME MEASURES: Proportion of admissions considered unnecessary or uncertain: agreement between observers on these proportions: effect of social and procedural factors on the admission decision. RESULTS: Between 8 and 34% of admissions were considered unnecessary and 20-38% of unclear necessity. Agreement between the groups of clinicians was not good. GPs and consultant surgeons showed the poorest agreement (kappa = 0.08 to 0.25, 4 comparisons), and the GPs scored a higher percentage of admissions as unnecessary (34 v 8-12%). After discussion, the consensus group achieved good to very good agreement (kappa 0.61-0.84). CONCLUSIONS: Different groups of doctors vary widely in their views about the need for emergency surgical admission. Good agreement can be reached by consensus discussion. GPs are less likely than surgeons to consider emergency surgical admission necessary. PMID- 9229064 TI - Prospective versus retrospective measurement of change in health status: a community based study in Geneva, Switzerland. AB - STUDY OBJECTIVES: To compare prospective and retrospective measurements of change in health status. DESIGN: Health status was measured using a French language version of the short form 36 (SF-36) health survey on two occasions one year apart--in 1992 and 1993. Differences in SF-36 scores measured prospectively were compared with the patients' single item retrospective evaluation of change in health (transition item). SETTING: This was a community based study among members of two health insurance plans in Geneva, Switzerland. PARTICIPANTS: Altogether 831 young adults (mean age 30 years at baseline). MAIN RESULTS: Health status remained stable on average during the study period. The retrospective rating correlated well with changes in health measured prospectively: those who said in 1993 that their current health was "much worse" than in 1992 experienced an average decrease of 1.06 SD on the eight SF-36 scales, while those who said that their health was "much better" recorded an average improvement of 0.43 SD. The associations between prospective and retrospective assessments of change were approximately linear for all scales but physical functioning. The transition item also discriminated between time periods: transition reported for 1991-92 did not correlate with changes recorded for 1992-93. Relative validity analyses indicated that the transition item was better suited to capture changes in general health than changes in purely physical or mental aspects of health. CONCLUSIONS: The concordance between retrospective and prospective measures of change in health suggests that both are sensitive, to some extent, to true changes in health status. Using both types of assessment may improve the reliability of measurements of change. PMID- 9229065 TI - Validation of a short telephone administered questionnaire to evaluate dietary interventions in low income communities in Montreal, Canada. AB - OBJECTIVE: To validate an adaptation of a short questionnaire measuring behaviour related to selecting low fat diets. The questionnaire was adapted for telephone use in a low income, low education population. DESIGN: The factorial structure of the 38 item adaptation was studied in a population based random sample of 1432 adults. Seven day test-retest reliability was measured in a convenience sample of 93 adults, and criterion related validity in measuring fat was assessed against a dietitian administered diet history in another convenience sample of 81 adults. SETTING: Adults aged 18-65 years living in low income, inner city neighbourhoods in Montreal, Canada. RESULTS: Principal components analysis identified five food factors: avoid fat, junk food, high fat traditional foods, low fat substitutes for high fat foods, and modification of meat to reduce fat. Two factors were similar to those of the original version. Internal consistency of the subscales ranged from 0.49-0.72. Test-retest reliability ranged from 0.72-0.90. Validation of the subscales against usual dietary intake indicated that the "junk food" factor, arising from questions added to the original questionnaire to reflect local dietary habits, was most closely related to fat intake (r = 0.48; p < 0.001). CONCLUSION: This telephone adaptation provides an inexpensive and valid method of measuring fat intake. However, these results suggest that adaptations of existing dietary instruments should be validated in the populations for which they are intended before they are used. PMID- 9229066 TI - The immediate effects of the pill safety scare on usage of combined oral contraceptives in north east England. PMID- 9229067 TI - Cause specific social class mortality differentials for child injury and poisoning in England and Wales. PMID- 9229068 TI - Epidemiology of neonatal tetanus in the Rivers State of Nigeria: a community based study. PMID- 9229069 TI - Variability of skin cancer registration practice in the United Kingdom. PMID- 9229070 TI - Research into purchasing health care: time to face the challenge. PMID- 9229071 TI - Discounting the future: influence of the economic model. PMID- 9229072 TI - Ionizing radiation and offspring sex ratio. PMID- 9229073 TI - ()-form distribution seen in microvascular cast specimens of the filiform and fungiform papillae on the anterior central dorsal surface of the cat tongue.) AB - Three-dimensional structures of the microvascular network of the filiform papillae (FiP) and fungiform papillae (FuP) on the cat tongue were observed by the corrosion cast method under a scanning electron microscope (SEM). FiP can be classified into five types, types I-V and FuP into four types, types I-IV according to the shape and size of the main process (MP) and the number of the accessory processes (AP) which are found on the anterior central dorsal surface. FuP (I-IV) were found to be distributed sporadically among FiP (I-V). Each of the types I-V of FiP were arranged the form of a in an oblique line running from the anterior central zone to the posterior peripheral zone in an orderly and geometrical fashion. Each of the types I-IV of FuP were scattered throughout in line of FiPs arranged in the form of a , the point of which is directed at right angles towards the apex. FiP play an important role in the drinking of milk and water, holding, masticating and swallowing the food and, after mixing the food with saliva, in the transporting of the food mass towards the pharynx. The MP of FuP was considered to be a modified form of the MP of FiP of the cat tongue and to function as part of a sense organ for taste. PMID- 9229074 TI - Angioarchitectural comparison of the fungiform papillae of the cat and rabbit using scanning electron microscopic specimens. AB - The functional and morphological characteristics of the fungiform papillae (FuP) on the anterior dorsal surface of the cat and rabbit tongues were studied and compared using a scanning electron microscope (SEM), because the comparison of the functional and morphological relationship of FuP in microvascular cast specimens (MVCS) between these tongues is as yet not clear. In both species, FuP were found to be distributed sporadically among the numerous filiform papillae (FiP). In the cat, in particular, FuP were classified into four types (FuP I-IV) according to the shape and size of a main process (MP) and the number of accessory processes (AP). Each FuP, (FuP I-III lay along oblique lines of FiPs) contained a MP and many APs lying at the anterior basal margin of the MP, whereas FuP IV contained only the small fishnet-ball-shaped MP. In the rabbit, however, only the similar size and shape of the capillary loops, resembling the carnation flower, were scattered among FiPs on the anterior dorsal surface of the fore tongue. These results suggest that in comparing the functional sense mechanism in both species, the FuP of the rabbit tend to be more simple than those of the cat in respect to taste sense. PMID- 9229075 TI - The ultrastructure and computer imaging of the lymphatic stomata in the human pelvic peritoneum. AB - The lymphatic stomata in the pelvic peritoneum of human fetuses and mature mice were initially observed and studied quantitatively by using computer image processing (C.I.P.) attached to a scanning electron microscope (SEM). Two types of mesothelial cells were found in the pelvic peritoneum of human fetuses and mature mice, i.e. flattened and cuboidal cells. The lymphatic stomata, arranged in clusters, were only found irregularly distributed among the cuboidal cells. The divergence of stoma area in the pelvic peritoneum of human fetuses varied greatly, ranging from 0.8 micron2 to 43.4 microns2. The average area of the lymphatic stomata in human fetuses was 10.00 +/- 9.44 microns2. The variation coefficient was 94.40. The standard deviations and standard errors were 9.44 and 0.98 respectively. Most of the lymphatic stomata in human fetuses were between 1.34 microns2 and 32.11 microns2 in size (accounting for 90%), with maximum and minimum values of 43.4 microns2 and 0.8 micron2. The average distribution density of the lymphatic stomata in human fetuses was 7.2% and the maximum density was 11.6%, which means that the average and the maximum absorption rates of the human pelvic peritoneum from the peritoneal cavity were 7.2% and 11.6% respectively. Therefore, it is suggested that the lymphatic stomata in pelvic peritoneum play an important role in draining materials from the peritoneal cavity, and that the absorption effect of the pelvic peritoneum is similar to that of the diaphragmatic peritoneum. PMID- 9229076 TI - Ultrastructural demonstration of constitutive nitric oxide synthase (cNOS) in neocortical glial cells and glial perisynaptic sheaths. AB - We investigated the localization of the constitutive isoform of nitric oxide synthase (cNOS) in the rat visual cortex by electron microscopy, using a pre embedding immunohistochemical method. NOS-immunoreactivity was demonstrated in very few somata, dendrites and axon terminals of nerve cells, and also in a few glial cells. The morphological characteristics of labelled glial cells enabled us to identify them as astrocytes. In comparison to the enzyme activity in neurons, the glial cells show a very weak reactivity which was not detectable at the light microscopical level. Immunoreactivity was located in the perikaryon and in the processes of astrocytes. Qualitative analyses with the electron microscope indicate that NOS-positive astroglial cells constitute a very small proportion in the rat visual cortex, whereas the great majority of astrocytes is NOS-negative. An unexpected observation was the localization of NOS immunoreactivity in astroglial sheath-like structures around some synaptic junctions. These results suggest that NO may act very locally at some synapses, and that specialized glial cells are involved in the NO-mediated mechanisms. PMID- 9229077 TI - Blood supply to the retina in the laboratory shrew (Suncus murinus). AB - The blood supply to both retinae was studied light microscopically and by scanning electron microscopy in 48 adult laboratory shrews (Suncus murinus) of both sexes. Thirty-eight of the animals were injected into the left ventricle with Neoprene latex (Du Pont. 601A) or with Mercox (Dai Nippon Ink Ltd., CL-2R) to elucidate the blood supply to the retina from the ophthalmic artery. The remaining animals were kept for histological study of the retina. The central retinal artery, originating from the ophthalmic artery in the muscular part of the orbit, enters the optic nerve, passes through the optic disk together with the central retinal vein and penetrates the vitreous space (cavity of the eye) between the lens and the inner limiting membrane of the retina, where it divides into the dorsal, ventral, and caudal branches. Each branch, moreover, bifurcates into nasal and temporal arterioles and is distributed throughout the retina on the inner limiting membrane as far as the ciliary body and the lens. On the way they obliquely send small vessels through the inner limiting membrane into the outer plexiform layer of the retina. Their vascularization appears to correspond to the membrana vasculosa retinae found in teleosts, amphibia and reptiles. PMID- 9229078 TI - Morphological changes elicited in skeletal muscle by a Nd:YAG laser scalpel and electrocautery during surgical reduction of the human tongue. AB - A Nd:YAG laser scalpel was used for the surgical reduction of a human hyperplastic tongue. This instrument combines a fine cutting precision with haemostatic properties, whereby loss of blood is minimized and the surgeon's field of view unimpeded by flooding from the damaged capillary bed. The coagulative properties of Nd:YAG laser light are, however, insufficient to effect blood flow stasis in larger calibre vessels (arteries > 2 mm; veins > 3-5 mm), such as those located at the base of the tongue. For this purpose, bipolar diathermy (electrocautery) was employed. The ultrastructural changes incurred by skeletal muscle fibres using these two "heat" sources were compared. The damage profile elicited using each modality was similar: coagulation of myofilamentous proteins leads to destruction of fibrillar architecture with concomitant loss of periodic banding; on moving away from the wound margin, characteristic features are gradually restored. As the severity of these heat-induced effects decreases, there is a corresponding increase in superimposed dislocation and tearing phenomena induced by post-treatment swelling. PMID- 9229079 TI - Effects of DHT and EGF on human hyperplastic prostate cells cultured in vitro: growth, morphology and phenotype characterisation. AB - This work studies the effects of dihydrotestosterone (DHT) and epidermal growth factor (EGF) on the growth, morphology and phenotype characterisation of the U285 line obtained from human prostate hyperplastic tissue. Modifications of growth rate induced by these two substances have been evaluated by means of the neutral red assay formulated by Borenfreund and Puerner (1985) as well as by means of Kenacid blue assay described by Knox et al. (1986), culturing cells for 24, 48 and 72 hr with scalar doses of DHT (0.5, 1, 2, 5, 10 microM) and EGF (5, 10, 20, 100 ng/ml). An optical microscope connected to a computer aided system and a scanning electron microscope were used to monitor morphological changes induced by DHT and EGF. The immunophenotype characterisation of the treated and control cells was carried out by using a monoclonal antibody panel. Our results show that the expression of anti-cytokeratin 5+6+18, anti-cytokeratin 8+18+19 and anti proline-4-hydroxylase antibodies varied in relation to the type of treatment undergone by the cells. Moreover, exogenous DHT does provoke a flattening of the U285 cells without modifying their rate of growth, while EGF both shortens the lag phase reactivating the quiescent cells and regulates the subsequent log growth phase, thus causing no cellular overgrowth. PMID- 9229080 TI - Venous drainage of the stomach in the laboratory mouse (Mus musculus v. alba) and the laboratory rat (Rattus norvegicus v. alba). AB - The authors studied the venous drainage pattern of the stomach in 30 adult laboratory mice (Mus musculus v. alba) and in 31 adult laboratory rats (Rattus norvegicus v. alba) of both sexes. In mice, two basic patterns of the venous drainage of the stomach have been found, the first one (50.0% of cases) with a vena gastroepiploica dextra, while in the second pattern (50.0% of cases) the vena gastroepiploica dextra is absent and the venous blood from the curvatura major ventriculi and fundus ventriculi is drained only via the v. lienalis. In rats, three basic patterns of venous drainage of the stomach were found, the first group (35.2%) with the v. gastroepiploica dextra, the second group (38.4%) with prevalence of the v. gastrica sinistra and the third group with various tributaries of the v. lienalis from the stomach (25.6%). The vena gastrica sinistra is the only constant venous channel in both animals examined. Between interorganic venous anastomoses in the mouse and the rat no great differences exist. In spite of the great variability of veins the results indicate that it is possible to differentiate some basic patterns of the venous drainage of the stomach in the animals studied. PMID- 9229081 TI - On a complex arrangement of the vascular pedicle of the left kidney. AB - During the dissection of an 83 year-old male cadaver, we observed that the left renal vein passed behind the aorta to follow an oblique course before draining into the inferior vena cava. The left renal vein was connected to the left suprarenal, spermatic, lumbar ascending and azygos veins. The left spermatic vein, after collecting the suprarenal vein, crossed in front of the abdominal aorta to reach the inferior vena cava directly. Furthermore, the arterial pedicle of the left kidney was composed of two tortuous and intermingled vessels. Variations in the number and arrangement of the vessels terminating in the renal veins are not uncommon, but so complex a vascular arrangement as this has not to our knowledge been previously described. The ontogenetic explanation and clinico surgical implications of such a variation are reviewed and discussed. PMID- 9229082 TI - The medial axis branch point in the human mandible. AB - The medial axis method was applied to radio-cephalometric images of the mandible in 20 adults and 18 children and to panoramic X-ray images of these children, and also directly to 50 halves of dry mandibles. It was found that the location of the posterior branch point coincided almost invariably with the mandibular foramen/lingula. The foramen may be regarded as the posterior limit of the mandibular body, from which the condylar and coronoid processes branch off. The medial axis appears to reflect the developmental and functional anatomy of the human mandible. PMID- 9229083 TI - Morphological and morphometric features of the deformed cervical and caudal vertebrae in a new mutant knotty-tail (knt/knt) mouse. AB - Morphological and morphometric examinations were conducted on skeletons of knotty tail (knt/knt) mouse, a new autosomal recessive mutant. The knt/knt mice have short and knotty tails. The number of caudal vertebrae is reduced and their deformed caudal vertebrae show no parallelism between the epiphyseal planes and no osseous fusions. Morphological changes, except for the caudal vertebrae, are confined to the neural arches of the axis in knt/knt mice. As for the tail anomaly, the knt gene seems to have the same action as the tk gene, but knt/knt mice differ from other strains with respect to cervical morphology. The morphometry of the caudal vertebrae revealed that knt/knt mice have a discontinuous and lower ratio of width to length after their 6th caudal vertebra. The morphometry also revealed that knt/knt mice have 1) broadened cervical vertebrae in the transverse direction, 2) thickened ventral lamina of the 6th cervical vertebra and 3) shortened and broadened ventral tuberculum of the atlas. From these results, the knt/knt mouse was considered to be a new pleomorphic mutant. PMID- 9229084 TI - Concha nasoturbinalis in human adult? AB - In a macerated skull from a 20 year-old body we found, bilaterally, a variation corresponding to the nasoturbinal concha of quadrupeds. According to the data in the literature, the remnants of this concha may be named either "agger nasi" or "agger cell". These formations may impede the approach to the frontal sinus or the lacrimal sac, respectively. A nasoturbinal concha is extremely rare in adults and should be considered when approaching the frontal sinus. PMID- 9229085 TI - Connective tissue disease and other rheumatic conditions following breast implants in Denmark. AB - To investigate the risks of connective tissue diseases (CTDs) following breast implants we used the nationwide Danish Hospital Discharge Register (HDR) to identify 2,570 women who received breast implants, either for cosmetic reasons (N = 1,135) or for breast reconstruction (N = 1,435), between 1977 and 1992. Two additional cohorts of women having either breast reduction surgery (N = 7,071) or breast cancer without implants (N = 3,952) were identified for comparison. Observed-to-expected (O/E) cases of CTDs and other rheumatic conditions were calculated based on national hospital discharge rates. The calculated O/E ratio for definite CTDs was 1.1 (95% confidence interval [CI], 0.2-3.4) among women with cosmetic breast implants, and 1.3 (95% CI, 0.5-2.6) among women receiving implants for breast reconstruction. No CTD excesses were seen in the breast reduction or breast-cancer-without-implant cohorts. Statistically significant risks for muscular rheumatism (a nonspecific discharge diagnosis) were observed in all four patient cohorts: cosmetic (O/E ratio, 2.5; 95% CI, 1.7-3.6), breast reconstruction (O/E ratio, 2.5; 95% CI, 1.7-3.4), breast reduction (O/E ratio, 2.0; 95% CI, 1.6-2.3), and breast cancer without implants (O/E ratio, 1.4; 95% CI, 1.0-1.9). In conclusion, breast implants showed little association with definite CTDs. Breast surgery per so, however, was associated with an apparent increase in muscular rheumatism. PMID- 9229087 TI - Tension and flap advancement in the human scalp. AB - The aim of the present study was to evaluate quantitatively the change in stiffness of scalp flaps determined by increased loads of tension. Data were collected by stepwise loading 20 scalp flaps, created by a reversed-Y incision down to and through the galea aponeurotica together with undermining. In the layer between the galea and the pericranium, to within 1 cm of the external auditory canal. The biomechanical properties of the tested scalp flaps were significantly influenced by increased extents of tension. The tissue's stress response to displacement was visualized as a three-phase characteristic. Initially linear (indicative load range, 0 to 500 g), the scalp's compliance gradually reduced (indicative load range, 500 to 1,500 g) and eventually demonstrated an exponential stress/strain characteristic of rapidly increasing stiffness (indicative load range, 1,500 to 5,000 g). The Young's modulus, E, of the stress/strain curve was found to be equal to 49.2 g per millimeter. The obtained data suggest that a reasonable approach should consist of closing a scalp defect within the tension range of 500 to 1,500 g. This will take full advantage of the plasticity of the scalp flap. The gain obtained with a closing tension above this range would be minimal, with a presumably increased complication rate. PMID- 9229086 TI - An outcome analysis of 100 women after explantation of silicone gel breast implants. AB - A prospective outcome analysis was conducted on 100 consecutive women who requested explantation of their silicone gel breast implants from January 6, 1992 (the moratorium), through 1995. Eighteen patients were referred by rheumatologists with a diagnosis of autoimmune or rheumatic disease. Six had autoimmune disease (systemic lupus, 2 patients; rheumatoid arthritis, 2 patients; multiple sclerosis, 1 patient; and Raynaud's disease, 1 patient). Twelve had rheumatic disease (fibromyalgia, 10 patients; inflammatory arthritis, 2 patients). All of these 18 patients had developed symptoms of their disease after they had received implants. All 100 patients were extensively evaluated pre- and postoperatively by interviews, clinical assessment, and by assay of the following laboratory tests: rheumatoid factor, ESR, ANA, and anti-Ro/SSA, -La/SSP, -Sm, RNP, -double-stranded deoxyribonucleic acid, -Scl-70, -centromere, and cardiolipin. Patients were also evaluated by a questionnaire that was sent at a mean time of 2.7 years postexplantation (range, 1-5 years), which had a 75% response rate. Reasons for implants were augmentation, 75%; lifting, 11%; reconstruction, 12%; and congenital aplasia, 2%. The mean age at first implant was 28.9 years (range, 13-55 years) and at explantation was 41.5 years (range, 25 65 years). The mean duration of implantation was 12.0 years (range, 1-27 years). Thirty-six percent of the patients had undergone at least one closed capsulotomy and 54% at least one open capsulotomy. The mean reasons for explantation were suspected silicone-related health problems, 76%; suspected rupture, 59%; breast firmness, 36%; breast pain, 36%; and musculoskeletal pain, 23%. Before explantation 75% of the questionnaire respondees had lost some sensitivity in their nipples following their breast augmentation. In 36% of those 75 patients, that loss was almost complete. Loss of sensitivity was related to capsular contracture and to pain (p < 0.05). Following explantation there was significant improvement in nipple sensitivity in 38% of breasts in the 75 respondees. A total of 186 implants were removed. Fifty-seven percent had failed by rupturing or leaking. Only 3.2% demonstrated extravasation extracapsularly. Twenty-five percent of the capsules were calcified, demonstrating visible plaques of calcification on their inner surface. Forty-two percent were colonized by bacteria. The prevalence of class III-IV capsular contracture was 61% and it was related to implant location, duration in situ, and capsular calcification (p < 0.05), but not to capsular colonization or implant integrity (p > 0.05). Only 43 of the 100 patients elected to have saline implants inserted. Of the others, 56% felt that the shell of the saline implant could be associated with medical problems. The others felt that breast size was of minor importance to them at this time. There were few complications from the explantation procedure. Two "masses" were discovered-one was an occult carcinoma, the other a galactocele. There was one wound infection, which responded to antibiotics. Three patients developed decreased sensitivity and 3 developed increased breast pain. From the patient questionnaires, in those women who did not have saline implants inserted, 15% felt that their breast appearance was improved after explantation, 36% were "pleased," 33% were disappointed, and 13% felt "mutilated". In women who did have saline implants inserted, 18% felt that their breast appearance was now improved, 60% were "pleased," and 14% were disappointed, mainly because of wrinkling. At a mean time of 2.7 years (range, 1-5 years) after explantation, 45% of the 75 questionnaire respondees felt that their implants had caused permanent health problems and 56% felt that they had not been given adequate informed consent by their original surgeon (particularly regarding implant rupture and a possible relationship to medical disease). (ABSTRACT TRUNCATED) PMID- 9229088 TI - Total reconstruction of a partial-thickness upper eyelid defect with the expanded forehead flap. AB - This paper demonstrates examples of successful reconstruction of partial thickness eyelid defects using the expanded forehead flap. Two patients are presented: one in whom cicatrical ectropion was present and the other in whom there was an absence of ectropion. In both patients primary grafting was executed just after the injury was sustained. By the time the patients were referred for reconstruction, mismatch in the quality of the skin and scars along the border of the graft left the area of primary repair to be impaired aesthetically as well as functionally. Reconstruction of the upper eyelid using the expanded forehead flap resulted in excellent approximation of the native tissue. Aesthetic as well as functional capacities were considered in this approximation. The advantage of this technique is that it offers a larger amount of tissue without compromising the aesthetic or functional similarity to the native eyelid. PMID- 9229089 TI - Submental artery island flap for reconstruction of the lower and mid face. AB - Ideal reconstruction of facial defects should be accomplished by like tissue. The submental artery island flap has the same characteristics as facial tissue, consisting of thin, pliable tissue with a perfect color match. The flap can be manipulated in different configurations employing skin, the platysma, the rim of the mandible, and the anterior belly of the digastric muscle to be utilized in the reconstruction of complex defects. The pedicle of the flap is quite reliable and enables a wide range of applicability. We have used this versatile flap successfully for various defects in 14 patients and our results are presented. Ten of the 14 flaps consisted of the skin and platysma, two flaps also included the anterior belly of the digastric muscle, one flap was elevated with the rim of the mandible, and one flap consisted of skin, the platysma, bone and muscle. PMID- 9229090 TI - Median sternotomy wound complication: the effect of reconstruction on lung function. AB - The objective of the study was to evaluate the lung function of patients with median sternotomy wound complication during the early postmedian sternotomy period and to compare the long-term pulmonary effects of reconstruction using pectoralis major and rectus abdominis muscle flaps. The percentage of predicted, standardized forced vital capacity (FVC); the standardized forced expiratory volume in 1 second (FEV1), and FEV1/FVC ratios of 45 patients with a median sternotomy wound complication were evaluated before and at a mean time of 10.6 months after wound reconstruction. Both mean FVC and FEV1 increased after wound revision compared with the prereconstruction results (8.4% and 9.2% increase, respectively). Patients with painful chest wall movement had the worst (60%) mean FVC and FEV1 before reconstruction when compared with a nonpainful complication. Reconstruction with a muscle flap was followed by an increase of 8.6% and 7.3% in FEV1 and FVC, respectively, from prereconstruction results. However, long-term results indicate that these patients have a mild, restrictive impairment of their lung function tests (LFTs), with about 80% of the predicted FVC and FEV1. Among the muscle flaps, the best improvement and best long-term LFT results were after sternectomy and reconstruction with a pectoralis major muscle flap as compared with a rectus abdominis muscle flap. Sternectomy and reconstruction with a muscle flap is a well-tolerated procedure associated with improvement of lung function compared with prereconstruction values. A pectoralis major muscle flap should be the first choice for muscle flap reconstruction while a rectus abdominis muscle flap should be reserved only for patients with good LFTs before reconstruction. PMID- 9229091 TI - Abnormalities of the hook of the hamate in patients with carpal tunnel syndrome. AB - The hook of the hamate is an important landmark in endoscopic carpal tunnel release. We studied the incidence of abnormalities of the hook of the hamate in patients with carpal tunnel syndrome. One hundred thirty-one consecutive patients presenting with carpal tunnel syndrome underwent carpal tunnel radiography. Abnormalities of the hook of the hamate were present in 6 of 131 patients (4.6%). The hook of the hamate was aplastic in 3 of 131 patients (2.3%). A bipartite hook of the hamate was present in 1 patient (0.8%). A nonunion of an old fracture of the hook of the hamate was found in two patients (1.6%). The previously reported incidence of an aplastic hook of the hamate in the general population is 1 of 1,452 (0.06%). The odds ratio revealed a 34-fold increase in the risk of hamate abnormalities in the carpal tunnel population (p < 0.001). Hook of the hamate abnormalities may be more common than previously reported. PMID- 9229092 TI - An experimental study of small-joint compression arthrodesis. AB - Arthrodesis of the interphalangeal and metacarpophalangeal joints is a technically demanding procedure with significant failure rates. A method of compression arthrodesis that was developed by one of the authors (RWB) using a compression clamp and crossed Kirschner wires is reported. This technique has been used without complication in the successful arthrodesis of 125 consecutive interphalangeal and metacarpophalangeal fusions by two of the authors (RWB and JAIG). An in vivo model of small-joint arthrodesis was then developed using the rabbit humeroulnar joint to compare this method of compression clamp arthrodesis with the tension band technique. Biomechanical testing at both 2 and 8 weeks postoperatively showed compression clamp arthrodesis to compare favorably with the tension band technique. PMID- 9229093 TI - The extended, pedicled rectus abdominis free tissue transfer for head and neck reconstruction. AB - The rectus abdominis musculocutaneous free tissue transfer has become a mainstay of reconstruction for large defects in the head and neck. The length of the deep inferior epigastric vessel is traditionally accepted to be 8 to 10 cm from its origin to its entrance into the rectus muscle. This pedicle is usually not long enough for reconstruction of the upper midface, forehead, and cranial base if vascular anastomosis to the neck vessels is necessary. Vein grafts have been recommended under these circumstances. We have been able to extend the length of the vascular pedicle by intramuscular dissection of the lateral branch of the deep inferior epigastric artery and veins in 26 clinical cases. The dissection is carried along the posterior surface of the muscle, up to the first tendonous inscription. Intraoperative measurements of the pedicle length before (6.9 +/- 1.0 cm) and after (13.7 +/- 2.0 cm) dissection, as well as in 17 fresh cadavers (34 muscles), demonstrate that the pedicle length can be increased safely from 7.8 +/- 0.5 cm to 17.7 +/- 0.52 cm (range, 15.8 to 19.1 cm). This long-pedicled flap has been used successfully in 26 patients for reconstruction of different types of defects in the head and neck, without using vein grafts. PMID- 9229094 TI - The role of pedicled or free fibular grafts in knee arthrodesis. AB - Arthrodesis of the knee can be facilitated by using a vascularized fibular graft if a large skeletal defect encompasses the knee joint or if conventional attempts at fusion have been unsuccessful. Solitary or double fibular grafts incorporated as part of a free tissue transfer or an ipsilateral pedicled flap rotated proximally about the knee are acceptable alternatives. Either version requires the technical expertise well known to reconstructive microsurgeons, as emphasized in this review, as a familiarity with this donor site and the nuances of graft elevation are essential for success. PMID- 9229095 TI - Anatomic evaluation of the facial artery and vein using color Doppler ultrasonography. AB - Despite the importance of preoperative evaluation of small vessels in flap elevation, the conventional methods such as palpation or Doppler probes are technically insufficient. These two methods do not detect varying venous patterns that would be identified during the operation in a "trial-and-error" manner. In order to overcome this problem in the facial vessels, an anatomic study was done using color Doppler ultrasonography in 12 adults. The artery and the vein were located together at the lower border of the mandible. On the other hand, around the oral commissure, they were located apart from each other with variable distances and variable size of vein. This divergence of the facial vein from the artery is important information in the planning of axial pattern flaps. It is suggested that color Doppler ultrasonography is a useful tool for venous identification in every body region. PMID- 9229096 TI - Treatment of extensive hypomelanosis with autologous cultured epithelium. AB - Successful repigmentation was achieved in 6 patients with three types of hypomelanosis (vitiligo, piebaldism, and albinism) by transplantation of fresh, autologous cultured epithelium with melanocytes. A small piece of uninvolved skin was taken for cultivation from a site adjacent to the lesion. Epidermal cells were cultured according to Green's technique. The lesions were abraded superficially and autologous cultured epithelium was applied. The grafts with functional melanocytes took completely and the wounds healed with minimal scarring. Repigmentation was visible within 6 to 8 months. The skin color resembled the surrounding normal skin except in the albinistic patient, in whom the donor skin was taken from a hyperpigmented area. Histochemical examination revealed dopa-positive melanocytes 12 to 17 days after grafting in the basal layer of the epidermis and the dermis. These cells grew in the basal layer of the epidermis and the hair follicles. Melanistic granules were visible in the keratinocytes in 1.5 months. A normal number of dopa-positive melanocytes and melanistic granules were seen in approximately 8 months. Thus, the autologous cultured epithelial grafting procedure is a promising treatment for patients with hypomelanosis. PMID- 9229097 TI - Clinical features and outcome of patients admitted to the intensive care unit after plastic surgical procedures: implications for cost reduction and quality of care. AB - Recent interest in cutting cost and improving utilization and delivery of perioperative services has prompted surgeons to identify patient populations that would benefit from care in an intensive care unit as opposed to intermediate or standard care. The purpose of this study was to evaluate patients admitted to the surgical intensive care unit (SICU) after major plastic/reconstructive surgical procedures in order to determine appropriate perioperative management strategies for these patients. We reviewed retrospectively the data from 2,805 consecutive admissions to the SICU between 1990 and 1996. Forty-two patients (1.5%) who had undergone major plastic/reconstructive procedures were identified. Outcomes (mortality, length of stay in the SICU and hospital, and the degree of organ dysfunction) were compared between this population, an illness severity-matched (Acute Physiology and Chronic Health Evaluation [APACHE]-II and APACHE III) population of patients recovering from vascular surgical procedures, and a similarly matched population of SICU patients who were randomly assigned to serve as a second control group. The hospital mortality of the plastic surgical patient population (9.5%) was significantly higher than the zero mortality of the random cohort (p < 0.05). A second analysis compared the SICU plastics group to a case controlled group of patients who were admitted to the postanesthesia care unit (PACU) for at least 24 hours of perioperative monitoring. SICU patients had significantly higher APACHE II scores (10.9) when compared to PACU patients (7.2; p < 0.01). Based on severity of illness scoring and eventual mortality, patients admitted to our SICU after major reconstructive surgery were selected appropriately for that setting. In contrast, the patients who stayed in the PACU for perioperative monitoring did not require life-supporting therapy and, therefore, were overmonitored. Care could be provided in a specialized unit with dedicated nursing specifically trained for that purpose. PMID- 9229098 TI - Mycobacterium smegmatis infection in a healthy woman following a facelift: case report and review of the literature. AB - In a 35-year-old female HIV-negative patient a facelift was followed by a Pseudomonas aeruginosa wound infection. The infection persisted despite treatment with ciprofloxacin, and an additional bacteriological wound examination revealed Mycobacterium smegmatis as the causative agent. Combination therapy with ciprofloxacin, doxycycline and amikacin led to a slow healing process without the need for further surgical intervention. A relapse 6 months after initial therapy was successfully treated with local measures. Infection with M. smegmatis might have come about by contaminated intraoperative liquids or the application of lipid creams to the open wound. However, microbiological examination of potential sources remained negative. Infection caused by M. smegmatis following plastic surgery should be considered in patients with spontaneous ulceration and violaceous discoloration of the skin adjacent to the surgical wound. Prolonged antibiotic therapy and possibly repeat surgical interventions may become necessary to treat this rare infection. PMID- 9229099 TI - Uncomplicated pregnancy following total bilateral rectus harvest: a case report. AB - Muscle-sparing rectus abdominis muscle harvest has been advocated to preserve abdominal wall integrity and to reduce postoperative complications and functional loss. In previous cases of pregnancy following single and bilateral rectus muscle harvest, muscle-sparing techniques were used to fortify the abdominal wall. We report a case of uncomplicated gestation and delivery following total bilateral rectus muscle harvest. While abdominal wall strength is greatly affected by rectus harvest, the fascial and tendonous components of the abdominal wall contribute substantial static integrity and elasticity. Careful repair of the donor site makes complete rectus muscle harvest an option even for women contemplating pregnancy. PMID- 9229100 TI - Maxillary myxoma treated with wide resection and immediate reconstruction: a case report. AB - Myxoma of the jaw is a rare, benign bone tumor of odontogenic origin. Given the locally aggressive nature of the myxoma and the high rate of recurrence, there is a tendency to choose radical resection of the jaw segment containing the tumor mass as the treatment modality. As they appear to attain a considerable size prior to diagnosis because of their insidious growth characteristics, particularly in the maxilla, treatment requiring a hemimaxillectomy often results in a significant functional and aesthetic disability. This is particularly distressing as this condition usually affects the young. We present the case of a 16-year-old female who suffered a large maxillary myxoma with swelling of the left cheek for 2 years. Wide resection of the tumor tissue including the maxillary medial and lateral buttresses and part of the hard palate with preservation of maxillary alveolar bone and teeth was undertaken, and the maxillary buttresses were reconstructed with autogenous rib grafts. At the 3.5 year follow-up there was no local recurrence and no tooth loss. Both the functional result and aesthetic contour proved satisfactory. Wide resection with preservation of vital structures and simultaneous autogenous bone graft reconstruction is our preferred method. A long-term follow-up is needed. PMID- 9229101 TI - Successful hand revascularization with urokinase following a crush injury. AB - Acute hand ischemia is a medical emergency requiring immediate treatment. We report a case of acute hand ischemia due to a crush injury of the wrist. Management with urokinase was successful in reestablishing flow to the ulnar artery and the digital vessels. In the setting of acute trauma with extensive thrombosis of the vessels of the hand, thrombolytic therapy may offer a better treatment option than surgical exploration with bypass grafting. PMID- 9229102 TI - Lateral ray polydactyly: a case of duplicated metatarsal with normal phalanges. AB - Lateral ray polydactyly of the foot is morphologically classified on the basis of the external appearance and the anatomic patterns of bony structures. Classification is difficult when the level of duplication is not apparent. We encountered a case of a duplicated metatarsal with no duplication of phalanges. The anomaly was considered to be a case of polydactyly because there were supernumerous metatarsals. On the basis of embryological failure, limb malformations are classified into several categories, and polydactyly belongs to that of duplication. There are, however, some malformations that are difficult to classify clearly. We considered our case to be a result of duplication followed by fusion of the digits. PMID- 9229103 TI - Benign giant schwannoma located in the upper arm. AB - A 72-year-old-female presented with a giant schwannoma on the medial side of her right upper arm. Ultrasonographic and magnetic resonance imaging examinations showed that it was almost a totally cystic lesion. It was initially misdiagnosed as a hydatid cyst. After excision of the tumor, histopathological examination revealed that it was a schwannoma composed of two types of regions known as Antoni A and B regions. The tumor was 15 x 8 x 7 cm in size. There were no neurological sequelae after the operation. This is probably the biggest schwannoma of the upper extremity reported. PMID- 9229104 TI - Re: Hyperbaric oxygen therapy and free radical production: an experimental study in doxorubicin (adriamycin) extravasation injuries. PMID- 9229105 TI - Complications in laser transconjunctival lower blepharoplasty. PMID- 9229106 TI - Re: How to harvest a septal chondromucosal graft. PMID- 9229107 TI - The longest pedicle for a gastrocnemius flap! PMID- 9229108 TI - Homeobox genes and disease. AB - To date, not many disorders have been associated with homeobox genes, especially with those belonging to the HOX family. This is particularly surprising, considering the body of evidence accumulated for a role of these genes in the control of mammalian development. Recently, this situation has changed and some congenital or somatic defects have been demonstrated to involve mutations in homeobox genes of the HOX, EMX, PAX, and MSX families, as well as in other novel genes containing either a paired- or bicoid-type homeobox. PMID- 9229109 TI - Sox genes find their feet. AB - The identification of the mammalian testis-determining factor, SRY, led to the description of a new class of genes encoding transcription factors, the SOX gene family. SOX proteins display properties of both classical transcription factors and architectural components of chromatin. The dynamic and diverse patterns of expression of SOX genes and analysis of mutations in humans, mice and Drosophila suggest that SOX factors play key roles in decisions of cell fate during diverse developmental processes. PMID- 9229110 TI - RNA-binding proteins as regulators of gene expression. AB - A plethora of post-transcriptional mechanisms are involved in essential steps in the pathway of genetic information expression in eukaryotes. These processes are specified by cis-acting signals on RNAs and are mediated by specific trans-acting factors, including RNA-binding proteins and small complementary RNAs. Recent information has begun to define the molecular mechanisms by which RNA-binding proteins recognize specific RNA sequences and influence the processing and function of RNA molecules. PMID- 9229112 TI - DNA mismatch repair in mammals: role in disease and meiosis. AB - The understanding of mammalian mismatch repair (MMR) gene function has been accelerated as a result of progress on several fronts. First, the biochemical analysis of MMR has been advanced by the production of purified human MMR proteins which will eventually allow reconstitution of MMR activity in vitro. Second, a wealth of clinical studies on colon cancer patients have begun to allow correlations to be made among MMR mutations, tumor types, therapeutic approaches and clinical outcomes. Finally, new unexpected meiotic phenotypes have been associated with mutations in certain mouse MMR genes. PMID- 9229111 TI - DNA helicases in inherited human disorders. AB - Six known or predicted helicases that are mutated in human syndromes are now recognized. These syndromes include xeroderma pigmentosum, Cockayne's syndrome, trichothiodystrophy, Bloom's syndrome, Werner's syndrome, and alpha-thalassemia mental retardation on the X chromosome. The clinical abnormalities in these syndromes cover a broad spectrum, pointing to different cellular processes of DNA manipulation that are defective in these syndromes. PMID- 9229113 TI - Serine/threonine kinase receptors and ligands. AB - Serine/threonine receptors transduce signals for the TGF-beta family, several members of which, such as decapentaplegic and bone morphogenetic proteins, are involved in early patterning of the embryo. The gene encoding the anti-Mullerian hormone (AMH) receptor has recently been cloned; gene targeting produces the same effects as targeting of the AMH gene itself. Another divergent member of the TGF beta family, GDNF, signals through Ret, a tyrosine kinase receptor; binding to Ret requires the cooperation of GDNFR-alpha. The signal transduction pathway of serine/threonine receptors is now being intensively studied; the immunophilin FKBP-12 and MAD proteins are known to be involved. PMID- 9229114 TI - Skeletal disorders associated with fibroblast growth factor receptor mutations. AB - Mutations in three fibroblast growth factor receptor loci underlie several autosomal dominant skeletal disorders; these include dwarfism and various craniosynostosis syndromes affecting limb and craniofacial bone patterning. A functional analysis of several of these mutations has demonstrated that a constitutive activation of the receptor kinase is a common theme. PMID- 9229115 TI - The sulfatase gene family. AB - During the past few years, molecular analyses have provided important insights into the biochemistry and genetics of the sulfatase family of enzymes, identifying the molecular bases of inherited diseases caused by sulfatase deficiencies. New members of the sulfatase gene family have been identified in man and other species using a genomic approach. These include the gene encoding arylsulfatase E, which is involved in X-linked recessive chondrodysplasia punctata, a disorder of cartilage and bone development. Another important breakthrough has been the discovery of the biochemical basis of multiple sulfatase deficiency, an autosomal recessive disorder characterized by a severe of all sulfatase activities. These discoveries, together with the resolution of the crystallographic structure of sulfatases, have improved our understanding of the function and evolution of this fascinating family of enzymes. PMID- 9229116 TI - The plakin family: versatile organizers of cytoskeletal architecture. AB - Desmoplakin, plectin, bullous pemphigoid antigen 1 and envoplakin are four sequence-related proteins--recently named the plakin family--that localize to intermediate filaments and filament attachment sites at the plasma membrane. New interest in the plakins has been stimulated by the discoveries that they can link different cytoskeletal elements together and that loss of plakin function can cause diseases of the skin and other tissues. PMID- 9229117 TI - The genetics of obesity. AB - Despite the influence of obesity in predisposing to many diseases, and evidence for high heritability, efforts to identify human genes with major effects on bodyweight have not yet been successful. In contrast, remarkable progress has been made in the identification and characterization of the genes mutated in five monogenic mouse models of obesity. These genes have led to new insights into the etiology of obesity and provide promising targets for therapeutic intervention. PMID- 9229119 TI - Turn to the worm! AB - Caenorhabditis elegans will be the first multicellular animal to have its entire genome sequenced. This is not just good news for those currently working in the field, but also for those trying to understand the biology of more complex animals, including humans. C elegans is a relatively simple animal that is amenable to studies of genetics and developmental processes that are common to all animals, making this an attractive model in which to study basic processes that are altered in human disease. Powerful forward and reverse genetics mean that virtually any gene of interest can be studied at the functional level. PMID- 9229118 TI - From gene to screen with yeast. AB - With the complete sequence now available, the yeast genome project enters a post sequencing phase that will concentrate on a comprehensive determination of gene function. Novel techniques have been developed to undertake genome-wide functional analysis at the levels of phenotype, transcript and protein. These include techniques for the efficient deletion of individual genes while tagging the deletants with specific oligonucleotide signatures, as well as strategies to quantify the physiological effects of such deletions by comparing growth rates and metabolite profiles under a range of conditions. Comprehensive approaches to the study of gene expression include hybridization array technology to identify and quantify transcripts, and the exploitation of mass spectometry to identify proteins resolved by two-dimensional gel electrophoresis. Yeast presents opportunities for the discovery of new human medicines both via the recognition of functional homologies between human and yeast genes and by the use of yeast to express human coding sequences specifying potential drug targets. PMID- 9229121 TI - Genetics of disease. PMID- 9229120 TI - Computational gene discovery and human disease. AB - Bioinformatics is now an essential tool in many aspects of human molecular genetics research. Methods for the prediction of gene structure are essential components in genomic sequencing projects and provide the key to deriving protein sequence and locating intron/exon junctions. Sequence comparison and database searching are the pre-eminent approaches for predicting the likely biochemical function of new genes, although sequence profiles derived from families of aligned sequences have advantages in the detection of remote sequence relationships. The use of sequence database analysis for large-scale comparative analysis of genome sequence data from model organisms is emerging as the most important recent development in the application of bioinformatics methods for characterizing candidate disease genes. PMID- 9229122 TI - Brain and language: what a difference a decade makes. PMID- 9229123 TI - New insights into the immunogenetics of multiple sclerosis. AB - The genetic analysis of multiple sclerosis has been based mainly on candidate gene association studies in selected populations. However, the candidate gene approach, although straightforward, is problematic in diseases of unknown or partially known etiology. With the advent of genomic screening and novel statistical approaches, it is possible now to perform an efficient screen of the entire human genome to identify the genetic components of multiple sclerosis. PMID- 9229124 TI - Pathology and pathogenesis of demyelinating diseases. AB - Multiple sclerosis is an inflammatory demyelinating disease of the central nervous system of putative autoimmune origin. In the present review the hypothesis that autoimmunity against multiple different brain antigens can lead to T-cell mediated brain inflammation and that multiple different immunological mechanisms may be responsible for the destruction of myelin is highlighted. The multitude of possible pathogenetic mechanisms is reflected in multiple sclerosis patients by a broad spectrum of disease susceptibility genes and by a profound heterogeneity of pathology and immunopathogenesis of the lesions. PMID- 9229125 TI - Strategies for repair and remyelination in demyelinating diseases. AB - A variety of therapeutic approaches aimed towards promoting myelin repair in multiple sclerosis and in other demyelinating diseases are now on the brink of clinical implementation. In this article, the extensive experimental studies of the past 2 decades, and in particular the considerable progress made over the last 12 months, will be briefly reviewed. A number of hurdles remain, together with practical and ethical problems which may be more difficult to solve; nevertheless, increasing excitement is apparent in this novel therapeutic field, and the rapid progress emerging from these laboratory based studies justifies a cautious optimism towards the successful translation of remyelination strategies from experimental neurobiology to clinical neurology. PMID- 9229126 TI - Measurement of treatment efficacy and new trial results in multiple sclerosis. AB - The past 12 months have witnessed a considerable number of new publications of the results of phase I, II and III trials of new, putative disease modifying therapies in multiple sclerosis. Some important reports have appeared concerning the development and optimization of clinical and magnetic resonance imaging methods for measuring therapeutic outcome. A general sense of optimism continues to exist that there will be further progress in coming years in achieving more effective strategies for treating symptoms, and for preventing relapses and disease progression. PMID- 9229127 TI - Central nervous system white matter diseases other than multiple sclerosis. AB - The essential difference between classical multiple sclerosis and both Devic's neuromyelitis optica and acute disseminated encephalomyelitis are outlined. Advances in identifying the multiple gene mutations responsible for adrenoleukodystrophy and possible mechanisms for the genotypic/phenotypic variability are reviewed. PMID- 9229128 TI - Bench to bedside: what have we learnt recently about headache? AB - Headache is perhaps the commonest of human maladies and recent years have seen large advances in understanding the basic mechanisms of head pain. The anatomy and physiology of pain transmission within the cranium have been elucidated, and advances been made into the pathophysiology of migraine and to a lesser extent cluster headache. PMID- 9229129 TI - The quest for migraine genes. AB - Research into the genetics of migraine remains difficult because of the involvement of polygenetic and environmental factors. The discovery of the gene for familial hemiplegic migraine on chromosome 19p 13 is an important step forward. This brain specific P/Q-type calcium channel alpha 1-subunit gene opens new avenues for studying the genetics of migraine, the pathophysiology of the onset of migraine attacks and the development of novel specific prophylactic drugs. PMID- 9229130 TI - Imaging the brain of migraine sufferers. PMID- 9229131 TI - Diagnosis and epidemiology of pediatric migraine. AB - Pediatric migraine differs from adult migraine in its epidemiology and symptom profile. Recent studies demonstrate the need to revise diagnostic criteria for pediatric migraine, demonstrate its epidemiology and suggest that its prevalence is increasing. These studies support the need for clinical trials to assess the utility of emerging therapies for pediatric migraine. PMID- 9229132 TI - Acute migraine therapy: the newer drugs. AB - In 1996, our knowledge of acute antimigraine therapy expanded in three major areas. First, large surveys have confirmed the remarkable efficacy profile of sumatriptan in clinical practice. No satisfying clinical, pharmacokinetic or genetic explanations were found for its major shortcomings: nonresponders, headache recurrence and noncardiac chest symptoms. Second, the novel 5-HT1B/D agonists zolmitriptan (311C90), rizatriptan (MK-462), eletriptan (UK-116,044), avitriptan (BMS-180048) and alniditan (R091274) were all proved superior to placebo for attack treatment, but their advantages over sumatriptan are yet to be analysed in more detail. A higher lipophilicity explains (except for alniditan) their greater oral bioavailability and better central nervous system penetration. A central action now proved experimentally in animals and in humans for 5-HT1B/D agonists such as zolmitriptan may be advantageous for the antimigraine efficacy, but it could also increase sedation. Third, an endothelin (Ro470203, bosentan) and a neurokinin 1 (RPR100893) receptor antagonist were found to be ineffective in migraine. Both compounds are potent inhibitors of neurogenic plasma extravasation in rat dura mater, which might suggest that this pharmacological property does not necessarily predict efficacy in aborting migraine attacks. PMID- 9229133 TI - Inflammatory diseases overview. PMID- 9229134 TI - Neuropathogenesis of acquired immunodeficiency syndrome dementia. AB - During the past year progress has been made in our understanding of the pathogenesis of the dementia associated with the acquired immunodeficiency syndrome. As many as one-third of acquired immunodeficiency syndrome patients eventually develop this condition, and at present it remains only poorly or transiently treated by existing antiretroviral therapies which do not penetrate well into the central nervous system. The past year has witnessed further characterization of microglial/macrophage neurotoxins, increasing evidence for neuronal death by apoptosis, and a more quantitative search for viral products, surrogate markers, or magnetic resonance spectroscopic parameters of brain or cerebrospinal fluid, or both. An increased understanding that the mediation of neuronal injury is not by direct infection of neurons, but rather via a complex network of cytokines, excitotoxins, and free radical mechanisms triggered by human immunodeficiency virus-infected or immune-stimulated brain macrophages and astrocytes has led to the development of therapies that are administered adjunctively with antiretroviral drugs. Some of these potential new treatments have now entered clinical trials. PMID- 9229136 TI - Rabies. AB - Rabies is a complex disease. We still do not understand the mechanisms of clinically diverse furious and dumb types and its fatal course. Moreover clinical symptomatology, once believed to be unique, may be variable, particularly in those patients who develop disease after exposure to virus of the insectivorous or frugivorous bat origin. This review summarizes classic and nonclassic clinical features associated with canine and bat rabies variants and also atypical presentations of rabies survivors. Difference in cellular tropism either at the inoculation site or in the central nervous system or differences in route of spread, or both, may account for these discrepancies. Furthermore, these may affect different sets of neurotransmitters that in turn modulate variable neurobehavioural patterns and neuroendocrine-immune cascades. PMID- 9229137 TI - Neurocysticercosis. AB - Neurocysticercosis is a pleomorphic parasitic disease that may mimic almost any other condition affecting the central nervous system. Recent studies have focused on diagnostic strategies and treatment of the most severe forms of the disease. This review covers the novel contributions made in those areas. PMID- 9229135 TI - Brain injury in bacterial meningitis: therapeutic implications. AB - Recent in-vitro studies have improved our understanding of how bacteria interact with cerebral endothelial cells and cross the blood-brain barrier. Several animal studies using rat and rabbit models of bacterial meningitis have revealed mediators of inflammation that are believed to play a key role in secondary brain damage, including reactive oxygen species, nitric oxide, and excitatory amino acids. Treatment with free-radical scavengers, nitric oxide synthase inhibitors, excitatory amino acid antagonists, as well as the anti-inflammatory cytokine interleukin-10 was beneficial in experimental bacterial meningitis. Apart from dexamethasone these agents hold major promise for the adjunctive therapy of bacterial meningitis in clinical practice. PMID- 9229138 TI - Prion diseases. AB - Prion diseases are a group of disorders sharing clinical and pathological features. Many of the enigmas of these diseases have now yielded to the concerted effort to understand them and their unusual pathogenesis. The genetic backgrounds of the various diseases are being clarified at an impressive rate, but the cause of sporadic Creutzfeldt-Jakob disease, the most frequently occurring human prion disease, is still not understood at all. The mechanisms underlying the evolution of the disease, as well as the species barrier are better understood. PMID- 9229139 TI - Demyelinating diseases. PMID- 9229140 TI - Headache. PMID- 9229141 TI - Inflammatory diseases. PMID- 9229142 TI - Advances in the molecular genetics of gliomas. AB - Malignant gliomas are the most common primary tumors of the central nervous system but remain clinically intractable. This has engendered substantial efforts to elucidate the molecular genetic and biologic basis of glioma formation. This review focuses on recent discoveries of the genetic aberrations that occur during the progression of gliomas. These include those that inactivate tumor suppressor genes and activate oncogenes. The biologic consequences of such genetic changes on various tumor cell-host environment interactions are also described. PMID- 9229143 TI - Tumor cell invasion and angiogenesis in the central nervous system. AB - Tumor cell growth and invasion within the CNS imply complex interactions among malignant cells, neural cells, and endothelial cells. Various in vitro assays have been developed to study tumor cell invasion that includes the use of cocultures between tumor spheroids and organotypic cultures of normal brain tissue. Furthermore, various animal models have been developed to study biologic characteristics of brain tumors. At present, there is evidence that several growth factors are involved in both endothelial and tumor cell proliferation, whereas the interrelationship between glioma growth and invasion is less well established. It is also emerging that the process of invasion is characterized by dynamic interactions between the extracellular matrix, proteases, and specific cell surface receptors. The dissemination of tumor cells within the CNS also involves a passive component where single tumor cells may follow specific pathways mediated by the constant flow of cerebrospinal fluid. PMID- 9229145 TI - Advances and controversies in the management of childhood brain tumors. AB - Important studies have been reported in the past year on the biology and management of pediatric central nervous system tumors. Pediatric low-grade gliomas and adult gliomas do not share similar molecular genetic features. There may be a role for high-dose chemotherapy with bone marrow rescue in relapsed medulloblastoma and germinoma. Malignant gliomas in infants appear to be distinct entities compared with adult malignant gliomas. Further investigational studies are needed in ependymoma and diffuse pontine gliomas because current therapies have not resulted in improvements in outcomes. PMID- 9229144 TI - Histopathologic and immunohistochemical prognostic factors in malignant gliomas. AB - Histopathology and immunohistochemistry continue to be popular methods for predicting outcome in patients with malignant gliomas. This past year traditional histopathologic studies have stressed the importance of endothelial proliferation in the diagnosis of glioblastoma multiforme. Immunohistochemical proliferation markers, in particular MIB-1, may be useful in assessing oligodendroglioma behavior, whereas their role in malignant astrocytomas is less clear. Similarly, new studies on p53 and epidermal growth factor receptor immunohistochemistry in gliomas have demonstrated only limited predictive values. PMID- 9229146 TI - Economic considerations in the care of patients with head and neck malignancies. AB - In an era of cost-consciousness and managed care, quality concerns practice variability attributed to nonmedical factors, and growing attention to outcomes research, there is increasing interest in the economics of malignant disease. This review explores economic issues pertinent to the management of patients with head and neck malignancies. Using economic principles to evaluate medical practice does not uniformly mean that less money should be spent; rather, the intention is to optimize efficiency in the use of limited resources. Accordingly costs are best evaluated in the context of other outcomes of interest. The available economic literature for head and neck tumors is limited; it is often compromised by the use of facility charges as a proxy for true costs and the adoption of a truncated economic perspective. Given the potential health policy implications of such studies, their methodologies and results warrant careful scrutiny. Many opportunities exist for further research. PMID- 9229148 TI - Clinical and molecular aspects of squamous cell carcinoma of the head and neck in the nonsmoker and nondrinker. AB - Most molecular analyses of head and neck squamous cell carcinoma involve patients who have used tobacco and alcohol. The pathways involved in tumorigenesis in patients lacking these lifestyle risks may be quite different. Research involving rigorous epidemiologic and molecular methods is needed to identify the unique spectra of genetic alterations in subsets of the population. This review considers recent information on the accumulation of genetic changes in the typical head and neck squamous cell carcinoma population and several studies that attempt to analyze subsets of patients categorized by age, gender, and carcinogen exposure. Efforts to identify risk factors in nonsmoking and nondrinking head and neck squamous cell carcinoma patients are evaluated, including studies of family history, human papillomavirus, chromosome fragmentability, microsatellite instability, and carcinogen-metabolizing enzyme genotype. PMID- 9229147 TI - Biomarkers in head and neck carcinoma. AB - Head and neck squamous cell carcinoma arises from a clonal population of cells that accumulate many genetic alterations in a multistep process. Each of these alterations may give these cells a growth advantage to allow them to progress toward malignant transformation. These alterations preferentially appear in a different stage of the carcinogenesis. The identification and characterization of these changes may not only provide insights into tumor biology, but also provide markers for many potential clinical applications. Progress in the identification of new biomarkers and their applications over the past 2 years is reviewed. PMID- 9229150 TI - Mucosal protectants and their application for head and neck chemoirradiation. AB - Mucosal injury due to ionizing radiation or cytotoxic agents begins with damage to stem cells in the basal epithelium, progresses as these cells are depleted, and is complicated by inflammation and superimposed infection. Suppression of oral and pharyngeal infection, cytokine stimulation of neutrophils, and protection of basal epithelium by chemical or physical means have been developed to protect mucosa from the acute effects of chemoirradiation. Although no method has been capable of preventing mucosal injury or its inflammatory consequences, many have proven partially effective and are reviewed in this article. Introduction of mucosal protectants into clinical use is needed to reduce the morbidity of chemoirradiation and to enhance its effectiveness by dose intensification, accelerated delivery, and simultaneous use. PMID- 9229149 TI - Status of accelerated fractionation radiotherapy in head and neck squamous cell carcinomas. AB - Considerable interest has been shown in recent years about hyperfractionated and accelerated radiation therapy for head and neck squamous cell carcinomas. The first randomized trial showing an advantage for hyperfractionation in terms of tumor control was the European Organization for Research and Treatment of Cancer 22791 trial. More recently, accelerated radiotherapy has been tested in many studies showing that high total doses of radiation could be delivered in overall treatment time shorter than conventional radiation therapy. The benefit of accelerated radiation therapy has been reported in some recently completed randomized trials, which suggests that rapid repopulation of surviving tumor cells during radiation therapy is a major determinant to obtain cure in this type of cancer. PMID- 9229151 TI - Prostate cancer. AB - Prostate cancer accounted for over 41,000 deaths in the United States in 1996. Prostate-specific antigen (PSA) screening is capable of detecting prostate cancer and appears to detect cancers that are both clinically significant as well as organ-confined, and therefore potentially curable. The positive predictive value of PSA value has been increased by the use of the free-to-total PSA ratio. The early detection of a large number of nonpalpable tumors has mandated the development of new risk assessment schemas, which include nomograms and equations in which Gleason score, PSA, and clinical stage play a prominent role. Definitive answers to the question of watchful waiting versus intervention await conclusion of the prostate cancer intervention-versus-observation trial. For both radical prostatectomy and radiation therapy, one means of potentially reducing the risk of relapse is the use of androgen deprivation. Neoadjuvant androgen deprivation prior to surgery results in a lower incidence of positive surgical margins, but impact on survival is unknown. By contrast, the use of concurrent androgen deprivation appears to be associated with enhanced survival in patients treated with definitive radiotherapy. For good risk tumors, modem brachytherapy results in freedom from biochemical relapse rates similar to those observed with surgery and external beam radiation therapy. The best therapy for patients with positive margins or serologic progression, including radiation therapy, remains to be identified. The widespread availability of PSA testing has led to an empirically driven redefinition of advanced disease and includes patients with earlier stage disease in which primary treatment has failed. In these patients, debate remains as to whether combined androgen deprivation is superior to monotherapy. A comparison of flutamide with bicalutamide awaits maturation of survival data. The utility of antiandrogen withdrawal in patients with progressive disease despite androgen deprivation has been confirmed. Thereafter, second-line hormonal maneuvers may be appropriate. In patients with truly hormone refractory prostate cancer, a variety of nonhormonal agents, including estramustine-based therapy, suramin, mitoxantrone, and doxorubicin-based regimens have demonstrated activity and remain as options. PMID- 9229152 TI - Testis cancer. AB - Although the concept that transplacentally acting estrogen-mimicking chemicals damage fetal germ cells is still the most favored hypothesis to explain the link between declining sperm counts and rising testis cancer, there has been increasing recognition that other mechanisms may be contributing. With reports confirming the association between a sedentary lifestyle and rising incidence of testis cancer and a fourfold increased relative risk of delay in conception of more than 3 months found for those driving a vehicle for more than 3 hours a day, there is increasing recognition that heat may be one of the most important cofactors. The role of p53 in heat-mediated damage and deficiency of heat shock protein response of germ cells and germ cell cancer are providing increasing interest in the search for molecular mechanisms to explain the unique chemosensitivity of this group of tumors. Perhaps the most controversial report was the finding of mutations insufficient to block apoptosis in 67% of tumor p53 genes using a RNA-SSCP analysis, when only a quarter of them had mutations identified by conventional DNA sequencing. The acceptance that immunosuppression, whether HIV- or chemically induced, increases risk of germ cell cancer by 20 to 50 times that in the general population is perhaps the most important final confirmation of the immune-surveillance hypothesis, although there is no evidence as yet that it seriously worsens the chance of long-term cure in these patients. Further progress is being reported on the use of high-dose chemotherapy and stem cell transplants, although the risks of treatment-related mortality still restrict its use to second line treatment. The problem of patient consent to the increasing range of options for early-stage disease is something highlighted from reports over the past year that will undoubtedly be an important issue in the future. PMID- 9229153 TI - Bladder cancer. AB - One of the most significant developments in the diagnosis and screening of bladder cancer is to use molecular genetic techniques and various diagnostic tools in hopes to adjunct cystoscopy, which is still the gold standard in diagnosis, and to define patients who are to be assigned more aggressive therapy. Although these techniques cannot replace cystoscopy, the promising results from these studies indicate that at least the frequency of control cystoscopies in the follow-up of low-risk patients may be decreased by the help of these tools. Bladder reconstruction following nerve-sparing cystectomy may represent the best primary surgical approach for organ-confined invasive disease at the present time. It is not yet clear whether bladder preservation with chemoradiation protocols will yield similar long-term disease free rates as in surgical series. However, it may be overoptimistic to think that appropriate randomized trials will address this question in the near future because physicians seem to have strong prejudices for one approach or the other. PMID- 9229154 TI - Renal cell carcinoma. AB - Renal cell carcinoma remains a major challenge for urologic oncologists. Over the past year, progress has been made in understanding the molecular biology of this disease and in describing new prognostic factors for outcome following nephrectomy. Studies have better defined patients who might benefit from elective nephron-sparing procedures without increasing the risk of relapse. Combination immunotherapy with interleukin-2 (IL-2) and interferon-alpha appears to be more active than single agent therapy and may be most effective when combined with chemotherapy. No new active chemotherapeutic agents have been identified. Prospective randomized trials suggest that autologous tumor cell vaccines as currently used offer little therapeutic benefit. PMID- 9229155 TI - Pediatric genitourinary tumors. AB - Each year advances are made in the clinical evaluation and treatment of genitourinary tumors in children. Our understanding of cellular, molecular, and genetic processes in tumorigenesis is evolving rapidly. In addition, knowledge concerning long-term outcome and complications associated with current treatments is increasing. In this article, we review recent literature on pediatric genitourinary tumors, including Wilms' tumor, rhabdomyosarcoma, and testicular tumors. PMID- 9229156 TI - Sedation and anesthesia for pediatric bronchoscopy. AB - Bronchoscopic examination may be indicated to evaluate various parts of the airway including the larynx, the subglottic region, or the more peripheral aspects of the tracheobronchial tree. The chances of a successful examination may be increased by the appropriate use of sedation. The following article reviews the steps required prior to during, and after the provision of anesthesia for bronchoscopic examination of the airway. The essentials of the preoperative examination, requirements for intraoperative monitoring, and postoperative recovery are discussed. The various agents and techniques available for deep sedation and general anesthesia are reviewed. PMID- 9229157 TI - Acute respiratory distress syndrome in children. AB - The adult (acute) respiratory distress syndrome is a significant cause of morbidity in children. The mortality rates remain elevated, greater than 50%, and even greater than 80% in patients with underlying malignancies. The therapeutic interventions remain mainly supportive. Strategies of conventional mechanical ventilation are directed toward the use of high positive end-expiratory pressures, low positive inspiratory pressure, and permissive hypercapnia. High frequency oscillatory ventilation and tracheal insufflation are not yet used extensively, although they should contribute to less aggressive ventilation. Surfactant replacement, nitric oxide inhalation, and partial liquid ventilation seem to be promising technologies, but controlled clinical studies are necessary before their wide-spread use. Extracorporeal membrane oxygenation remains the alternative technology in case of failure of conventional support. PMID- 9229158 TI - Pulmonary fungal infections in immunocompromised children. AB - Pulmonary mycoses are increasingly encountered in children with underlying diseases associated with immunosuppression, including children with cancer or HIV infection. Knowledge of the epidemiology and pathogenesis of pulmonary mycoses can be applied to prevent infection in many cases and to promptly diagnose infections that do occur. Treatment is usually successful if initiated early, although pulmonary aspergillosis and zygomycosis are portentous ailments unless surgical resection or prompt immunologic recovery ensue. PMID- 9229159 TI - Pulmonary hemorrhage in infants and children. AB - Following a brief presentation of important clinical concepts regarding pulmonary hemorrhage in infants and children, recent reports on secondary and immune related disorders causing acute pulmonary hemorrhage are reviewed. Idiopathic pulmonary hemosiderosis is updated noting the compilation of Japanese cases and current treatments. The recent increase in idiopathic pulmonary hemorrhage and hemosiderosis in young infants, particularly in Cleveland, is discussed along with management suggestions and an apparent relationship to some sudden infant death syndrome cases. PMID- 9229160 TI - The role of the pediatrician in smoking prevention. AB - Smoking prevention during childhood and adolescence is critical to the successful reduction of tobacco-related morbidity and mortality in the United States. The research literature is replete with surveys describing youth smoking acquisition and related factors. In addition to its direct harm, tobacco use may be a gateway to other substance abuse and a marker of other health-compromising behaviors. The pediatrician can play an important role in prevention by screening all patients and providing individual counseling. The pediatrician can also contribute as a consultant and advocate of school- and community-based smoking prevention efforts. Such a multicomponent approach provides the greatest likelihood of accelerating the decline in smoking and smokeless tobacco initiation. PMID- 9229161 TI - Acute hypertensive crises in children: emergencies and urgencies. AB - Hypertensive crises result from acute elevations in blood pressure. Although uncommon in children, they can be life-threatening and require immediate recognition and treatment to decrease morbidity. The diagnosis is made following complete history and physical assessment, with confirmation of blood pressure elevation using an appropriate-size blood pressure cuff. There are various options for treatment. Some of the more commonly used agents include nitroprusside, diazoxide, hydralazine, labetalol, esmolol, nicardipine, nifedipine, enalaprilat, and minoxidil. Close monitoring of blood pressure during treatment is mandatory to ensure a good outcome. PMID- 9229162 TI - Management of the suicidal child or adolescent in the emergency department. AB - The pediatrician may be called on either as the primary physician or as a consultant to see a suicidal child or adolescent in the emergency department. Psychiatric consultation may not be available immediately. The pediatrician will then play a pivotal role in the evaluation and disposition of the patient. We discuss the epidemiology, initial approach to evaluation, and a rational plan for deciding when it is safe to discharge the patient from the emergency department. PMID- 9229163 TI - Emergency management of oral trauma in children. AB - Oral trauma continues to be a common pediatric emergency, accounting for 150 emergency room dental consultations per year at Children's Hospital in Boston. Children between the ages of 18 months and 2.5 years and between 8 and 11 years are most at risk. Recent advances in the management of these dental emergencies may help children and their families avoid the psychological and financial cost of infection or loss of primary and permanent teeth. Treatment of avulsions in the young permanent dentition remains a common problem, and a universally accepted approach to its management is still evolving. The use of a doxycycline immersion prior to reimplantation by the dentist may be helpful in preventing external root resorption. As always, the best therapy against dentofacial trauma is the pediatrician's support of preventive measures. PMID- 9229164 TI - Pain management in the pediatric intensive care unit. AB - Critically ill pediatric patients are frequently exposed to acute, established, and chronic pain as a result of their disease processes or intensive care therapies. Despite the availability of many drugs and techniques for providing analgesia, these painful conditions are not adequately treated in a large proportion of children. This article reviews some of the reasons for provision of adequate analgesia and sedation, describes the various classes of drugs commonly used in the pediatric intensive care unit, and lists the techniques and indications for regional and topical anesthesia as well as specific clinical applications for adjuvant analgesic agents. Analgesic approaches that do not have an established record of safety and efficacy in pediatric patients are not reviewed. We propose that adequate and early analgesic interventions will minimize patient's discomfort, maintain metabolic homeostasis, and improve a patient's tolerance of intensive care unit therapies and nursing interventions. Adequate analgesia can be provided to even the sickest child using the drugs, techniques, and novel approaches reviewed. PMID- 9229165 TI - Emergencies in international child health. AB - Emergencies in the pediatric populations of third world and developing countries are of a much different sort than those to which pediatricians in developing countries are familiar. Many of these emergencies derive from conditions, situations, and etiologies that no longer represent a threat to children in developed countries: malnutrition, immunizable illnesses, infectious diseases from pathogenes easily treated or prevented, urbanization, and armed conflict. Programs directed at improving basic public health, health education, access to basic health care, and immunization have been shown to have a major and positive impact on children's health status in these countries. Because of the vastness of these health problems, a growing number of volunteer organizations offer opportunities for pediatricians to contribute to improvement and they have an impact on the health of children considerably less fortunate than those in developed countries. PMID- 9229167 TI - Biliary atresia. AB - Biliary atresia, a progressive obliterative process involving the bile ducts, has its onset in the newborn period. It is characterized by worsening cholestasis, hepatic fibrosis, and cirrhosis, which lead to portal hypertension and a decline in hepatic synthetic function. Untreated, the outcome is uniformly fatal. The two major milestones toward improved treatment of this disease have been the Kasai portoenterostomy and orthotopic liver transplantation. There has been discussion regarding transplantation as primary therapy, but portoenterostomy remains the standard of care as first-line intervention. Hepatic transplantation, done more frequently for biliary atresia than for any other cause of liver failure in the pediatric population, offers improved survival and quality of life to those for whom the Kasai operation fails. The etiology of biliary atresia remains poorly understood. Working toward a better understanding of this disease, recent investigations target more precise characterization of the hepatic pathology and seek to identify possible causative agents and predictors of favorable outcome. Recent advances in the understanding of biliary atresia published between December 1995 and November 1996 are the focus of this review. PMID- 9229168 TI - Parenteral nutrition-associated cholestasis. AB - Parenteral nutrition-associated cholestasis is a persistent problem that has been a major cause of morbidity and mortality in young neonates. This review discusses some of the more recently associated risk factors, potential etiologies, including some potential genetic causes, and discusses potential forms of therapy, including the use of ursodeoxycholic acid and cholecystokinin. PMID- 9229166 TI - Acute pancreatitis. AB - The etiology of acute pancreatitis in children is widely varied and includes idiopathic, drug-related, congenital, and posttraumatic causes. Most children have abdominal pain and tenderness without evidence of peritonitis, and most patients will have elevated serum amylase levels initially or after a delay of about 12 hours. If the diagnosis remains equivocal or in the setting of trauma, an abdominal CT scan should be obtained. Initial treatment for all forms of acute pancreatitis includes bowel rest and support with intravenous fluids. A nasogastric tube should only be placed for symptomatic relief and prophylactic broad-spectrum antibiotics should be given only in the setting of necrotizing pancreatitis, especially if patients are receiving pharmacologic immunosuppression. Fever or decline in clinical status should prompt CT scan with intravenous contrast and possible fine needle aspiration to detect the presence of sterile or infected necrotizing pancreatitis. Positive cultures or severely worsening clinical status are indications for necrosectomy and debridement with sequential packing and explorations. All patients who have had an episode of gallstone pancreatitis should have a cholecystectomy after resolution of pancreatic inflammation but before discharge from the hospital. PMID- 9229169 TI - Cholelithiasis, cholecystitis, and common bile duct stones. AB - Cholecystitis and cholelithiasis are being recognized with increasing frequency in infancy, childhood, and adolescence. Hematologic disorders account for a large proportion of cases; however, in most cases the etiology is uncertain. Infants and children are noted with stones in association with total parenteral nutrition, prolonged fasting, or ileal resection. Biliary dyskinesia, a disorder of impaired gallbladder contractility, is being recognized with increased frequency in late childhood and teenage years. Spontaneous stone resolution is frequently noted in infancy, and a period of observation is appropriate in the absence of symptoms. Laparoscopic cholecystectomy is the procedure of choice for symptomatic cholelithiasis and biliary dyskinesia. Common bile duct stones are unusual in children, occurring in 2% to 6% of children with cholelithiasis, often in association with obstructive jaundice and pancreatitis. Endoscopic retrograde cholangiography with stone extraction performed before or after laparoscopic cholecystectomy is the procedure of choice in this setting. PMID- 9229170 TI - Choledochal cysts. AB - Choledochal cysts are a relatively rare abnormality in the West but are more common in the East. The etiology of choledochal cysts remains unknown. Recently, the incidence in neonates and young infants has been increasing due to advances in diagnostic imaging, including antenatal diagnosis. Choledochal cysts can present at any age, but the clinical manifestation differs according to the age of onset. Early diagnosis followed by cyst excision is the treatment of choice, even in asymptomatic children. Recently, attention has been paid to the treatment of intrahepatic and intrapancreatic ductal diseases such as intrahepatic duct dilatation, debris in the intrahepatic ducts, and protein plugs in the common channel. Intraoperative cyst endoscopy is strongly recommended as a valuable adjunct to cyst excision for the prevention of postoperative complications due to intrahepatic or intrapancreatic ductal diseases. PMID- 9229172 TI - Spondyloarthropathies. PMID- 9229171 TI - Infant nutrition: implication for somatic growth, adult onset diseases, and oral health. AB - The gold standard for assessing the adequacy of nutrient intake in pediatrics is that diet which promotes optimal growth and development. Thus, it is crucial that our methods for measuring these outcomes be valid, reliable, and widely accepted. A review of the recent medical literature in the field of clinical nutrition indicates that both growth data and dietary standards continue to evolve as more data accrue concerning their applicability in both health and disease. In addition, oral nutrition is clearly a determinant of perhaps the most prevalent infectious disease in pediatrics: dental caries. Research in this field stresses the importance of oral fluoride intake in the prevention of caries, as well as the fact that current efforts at reducing milk-bottle tooth decay are inadequate. PMID- 9229173 TI - Diagnostic and classification criteria, clinical and functional assessment, and therapeutic advances for spondyloarthropathies. AB - Two sets of criteria have been proposed and widely accepted in the last few years for the classification of the whole spectrum of spondyloarthropathy, including the undifferentiated forms. These classification criteria--the Amor criteria and the European Spondyloarthropathy Study Group criteria--are not, however, particularly helpful for diagnosis because they do not include the milder and monosymptomatic forms. Outcomes research in spondyloarthritis is growing, and new instruments have been suggested. An international study group of experts is working to propose a core set of measures to be included in future clinical trials on ankylosing spondylitis. Intrasynovial corticosteroid injections in the sacroiliac joints may represent a valid alternative for patients with inflammatory low back pain that is unresponsive to nonsteroidal anti-inflammatory drugs. Sulfasalazine is an effective therapy for the psoriatic arthritis and peripheral arthritis of ankylosing spondylitis. A recent study has suggested its efficacy in reactive arthritis as well. In reactive arthritis, the use of long term antibiotic therapy has been proposed and is under study. PMID- 9229174 TI - Clinical and therapeutic aspects of juvenile-onset spondyloarthropathies. AB - Relatively little is written about the juvenile spondyloarthropathies. The literature of the past year has included data on the frequency of juvenile spondyloarthropathies, which indicate that these are almost certainly a more common form of childhood arthropathy than formerly believed. Although clinical differences exist between juvenile spondyloarthropathies and juvenile rheumatoid arthritis, there is only limited information about differences in the pathophysiology of these diseases. One study suggests some differences in the expression of tumor necrosis factor and its receptors. Evidence also presented this year suggests that juvenile psoriatic arthritis is probably a separate condition from the spondyloarthropathies. It is hoped that better understanding of the epidemiology and pathophysiology of the juvenile spondyloarthropathies will lead to better treatment strategies. PMID- 9229175 TI - Clinical, immunopathogenic, and therapeutic aspects of psoriatic arthritis. AB - Psoriatic arthritis is a common chronic rheumatic disease that may result in considerable joint damage if left untreated. Recent clinical studies have focused on the identification of poor prognostic factors. Early, aggressive treatment is indicated in patients with significant joint inflammation and certain HLA antigens. Immunopathogenic evidence continues to point toward an HLA class I mediated disease with perhaps an important role for CD8+ T cells. Although few basic research publications discuss psoriatic arthritis, studies in the areas of spondyloarthropathy and psoriasis continue to shed light on disease mechanisms. Greater collaboration is advocated among researchers in these areas and those interested in achieving a better understanding of the disease pathogenesis of psoriatic arthritis. PMID- 9229176 TI - Pathogenic role of gut inflammation in the spondyloarthropathies. AB - Some of today's most exciting immunologic research relevant to the pathogenesis of spondyloarthropathies concerns the relationship between the host immune system in the gut and in the intestinal bacteria. Through transgenic and gene knockout studies in rodents and the development of appropriate experimental models, it has become clear that this relationship depends on a fine balance in the production of different cytokines. Surprisingly, HLA-B27, acting at the level of T-cell recognition, disturbs this balance in ways that promote the development of arthritis. Exploring the mechanism of this disturbance and the role of genes other than B27 promises to illuminate the connection between gut inflammation and spondyloarthropathy--which has been carefully documented by clinicians over the past 30 years. PMID- 9229177 TI - Predisposing factors to spondyloarthropathies. AB - Predisposition to ankylosing spondylitis is largely genetic, and epidemiologic studies suggest that the environmental trigger is ubiquitous. HLA-B27 and -B60 predispose to ankylosing spondylitis, but in neither case is the mechanism of effect known. Other major histocompatibility complex and non-major histocompatibility complex genes are likely to influence susceptibility to spondyloarthritis as well as the disease pattern. Spondyloarthritis occurs in genetically predisposed individuals exposed to certain as yet undefined environmental triggers. A though genes within the major histocompatibility complex are clearly major determinants of susceptibility to spondyloarthritis, epidemiologic evidence suggests that their contribution accounts for less than 50% of the total. The mechanism of association of B27 with these diseases is unknown; we are currently unable to predict which B27 carriers will develop arthritis or which form of B27-associated spondyloarthritis they will develop. Lessons from transgenic animal experiments and technical and statistical advances in the field of genetics have greatly increased our ability to investigate these questions. PMID- 9229178 TI - Septic bone and joint infections. AB - During the past few years, new information on the pathogenesis of skeletal and joint infections and the host-parasite interaction has been presented. Although this is basic research, several recent case reports have shown that experimental laboratory findings are often relevant for the practical treatment of patients. Bacteria long known as causative agents in orthopedic infections, as well as new and rare ones, challenge our skills in diagnosis and encourage new thinking in host-parasite interaction and epidemiology. Another expanding problem is increased resistance against antibiotics, which is now being observed in pneumococci causing joint infections. PMID- 9229179 TI - Reactive arthritis. AB - The complex interactions between the triggering microbe and the defense mechanisms of the host in reactive arthritis have been studied in several laboratories around the world, and interesting observations have been made. Research has also focused on the mediators in the inflammatory process in joints, and these results are helping to slowly build a comprehensive picture about the pathogenetic process in reactive arthritis. For the practicing clinician, some important new findings have emerged. It is known that asymptomatic urogenital infections are quite common as a trigger of reactive arthritis. More aggressive treatment, including disease-modifying drugs as used in the therapy of rheumatoid arthritis, is becoming accepted. The value of antibiotic treatment is being studied, but final conclusions will not be made for perhaps a few years. PMID- 9229180 TI - Lyme disease. AB - The number of reported cases of Lyme disease in the United States has increased steadily since 1982, and the increase coincides with a growth in deer populations in endemic regions. Application of careful epidemiologic principles has led to the identification of two new pathogens that produce syndromes closely related to Lyme disease. Coinfection with other pathogens such as Babesia has been shown to alter the clinical course of Lyme disease. Better understanding of the pathogenesis of Lyme disease has provided clues into the mechanisms responsible for persistent symptoms and will serve as a foundation for the design and implementation of appropriate therapic recommendations. A safe vaccine for the prevention of Lyme disease in humans has been developed, and clinical vaccine efficacy trials are close to completion. PMID- 9229181 TI - Viral arthritis. AB - Viral infections are important in rheumatic disease not only because of the acute and subacute syndromes they cause but also because of their potential importance as etiologic factors in common chronic diseases such as rheumatoid arthritis. During 1996, the clinical spectrum of the acute polyarthritis caused by parvovirus B19 was further delineated and was shown to include carpal tunnel syndrome, hepatic dysfunction, and possibly angioedema. B19 infection can be studied using the salivary antibody response. No convincing additional data were reported regarding B19 in chronic syndromes such as rheumatoid arthritis or Behcet's syndrome. Regarding rubella as a possible cause of chronic arthropathy, more negative evidence accumulated with two additional studies in vaccinees and chronic arthritis using epidemiologic and virologic methods including the polymerase chain reaction. To define a possible link between rubella and autoimmunity in vitro, interactions of rubella RNA, ribonucleoprotein complexes including Ro and La, and calreticulin were explored. There was an avalanche of new information about hepatitis C virus infection, particularly its relationship to mixed cryoglobulinemia and related clinical syndromes. These syndromes have become much more commonly recognized, particularly in areas of high prevalence of hepatitis C virus infection such as Italy. The lymphotrophic nature of the virus is probably ultimately responsible for the rheumatic disease manifestations. Treatment is still problematic, but immunosuppressive drugs should be avoided. Epstein-Barr virus appears to have an etiologic role in the lymphomas occurring in immunosuppressed patients, including those who have had methotrexate therapy. Significant new studies regarding other viruses did not appear during the past year. PMID- 9229182 TI - Paget's disease of bone. AB - The highest incidence of Paget's disease is in the United Kingdom and is lower in other European countries. Worldwide the incidence appears to be decreasing. Some countries have observed a reduction in the severity at presentation, perhaps reflecting changes in migration of the UK population. Evidence of a genetic susceptibility to Paget's disease is accumulating, with chromosome 6 (the HLA locus) and 18q emerging as strong candidates. Use of molecular biology techniques has yielded evidence on the possible role of paramyxovirus infection in the etiology of Paget's disease. Altered measles virus nucleocapsid transcripts have been detected in bone marrow cell cultures and peripheral blood mononuclear cells from Paget's disease patients. Canine distemper virus may be the etiologic agent, and ownership of a mongrel dog or a dog not vaccinated against canine distemper virus is associated with an increased risk of Paget's disease. Familial Paget's disease is associated with more severe disease than is seen in sporadic cases. Quality of life may be reduced in patients with Paget's disease, and psychological problems are often present. New biochemical markers of resorption and formation appear to offer little advantage over urinary hydroxyproline and serum total alkaline phosphatase measurements in assessing Paget's disease activity or response to therapy. The best therapeutic responses are observed following treatment with the bisphosphonates, and oral etidronate is now being superseded by oral tiludronate, oral or intravenous alendronate, and clodronate or intravenous pamidronate. PMID- 9229183 TI - Endocrine disorders and osteoporosis. AB - Endocrine disorders constitute the most frequent cause of secondary osteoporosis in men and women. Because endocrine diseases are common (e.g., diabetes mellitus, hyperparathyroidism, and hyperthyroidism), they should be considered in the differential diagnosis and management of osteopenia. The pathogenesis of hormone induced bone loss involves several components of the bone remodeling cycle and in many cases is not fully elucidated. Identifying an underlying cause and correcting hypo- or hyperfunction of an endocrine gland, however, can often lead to an increase in bone mineral density. This review focuses on recent studies on hormonal disorders that affect the skeleton. In particular, diabetes mellitus, hyperparathyroidism, and hyperthyroidism are considered within the context of both accelerated bone loss and fracture risk. PMID- 9229184 TI - Effect of estrogens on bone and other systems and hormonal substitute treatment. AB - This article reviews recent advances in the effects of estrogen on bone and other body systems. The effect of estrogen use for the prevention of osteoporosis is no longer a matter of debate. Some issues are still discussed, however, especially with regard to the magnitude of its protection (especially for hip fracture), which would be dependent on the time at which estrogen therapy is initiated with respect to menopause as well as its total duration of use. The mechanisms of estrogen's action on bone also remains poorly understood. Controversy persists concerning the specific contribution of an indirect action through the modulation of calciotropic hormones and a direct action on bone cells in which estrogen receptors have been identified. Estrogens have been shown to reduce osteoclastogenesis by modulating the production of cytokines (interleukins, tumor necrosis factor) and transforming growth factor from bone marrow and bone cells. In addition to its effects on bone, a great number of observational studies have evaluated the relationship between estrogen use and coronary heart disease. The overall risk of coronary disease is estimated to be reduced by 50% in women who use estrogen replacement therapy. This reduction would be even greater in women with previous atherosclerosis. Moreover, recent data suggest that estrogen use could be associated with delayed onset and reduced risk for Alzheimer's disease. Overall, estrogen treatment in postmenopausal women has several proven benefits. Conversely, the potential risks of long-term estrogen therapy include a slightly increased risk of breast cancer, which would be associated with long-term use. Although the individual decision for a postmenopausal woman to start estrogen replacement therapy must take into account this risk:benefit ratio, current evidence suggests a clear advantage for estrogen's benefits over its risks. PMID- 9229185 TI - Diagnostic and therapeutic aspects of calciotropic hormones. AB - The measurement of calciotropic hormones may be useful in metabolic bone disease. Assays of intact parathyroid hormone are essential to differentiate between primary hyperparathyroidism and nonparathyroid-mediated hypercalcemia. Vitamin D status is best assessed by measuring serum 25(OH)D. Concentrations of calciotropic hormones should be measured in osteoporotic patients if the degree of osteopenia does not correlate with the risk factors. The decrease in circulating parathyroid hormone in osteoporotic patients treated with vitamin D reflects the success of the vitamin D treatment. Parathyroid hormone is also a potential anabolic agent. PMID- 9229186 TI - Spondyloarthropathies. PMID- 9229187 TI - Infectious arthritis and immune dysfunction. PMID- 9229188 TI - Metabolic bone disease. PMID- 9229189 TI - Human insulin allergy-immediate and late type III reactions in a long-standing IDDM patient. AB - Human insulin allergy-immediate or late type III reaction-is a rare event. We report the case of a 33-year-old female patient with insulin-dependent diabetes mellitus for 25 years who presented, in the last 8 years, mild but generalized urticaria partially controlled with oral antihistamines. There was no improvement after changing from mixed beef-pork to human insulin. In the last 3 years another allergic manifestation began: small, localized, subdermal and painful non erythematous nodules with central hematomas at injection sites, occurring 6-8 h after the insulin injection and lasting for 48 h. The following maneuvers had no benefit: (1) Human insulin (NPH or Lente) administered with dexametasone or xylocain locally, (2) Short acting human insulin with or without previous boiling, (3) Anti-histamine cetirizine dihydrochloride-10 mg/day. The allergic symptoms disappeared only after treatment with short acting human insulin (up to 100 U/day) associated to prednisone-40 mg/day and cetirizine dihydrochloride for 4 months. However, after stopping prednisone the urticaria reappeared and it was relieved with insulin desensitization. The pain at the site of injections persisted. CONCLUSION: This long-standing IDDM patient presented two types of reactions to human insulin: the immediate type (systemic urticaria), treated with antihistamines and desensitization, and the Arthus' type III reaction (nodules and hematomas occurring 6-8 h after the insulin injection) that required glucocorticoid therapy for more than 4 months. PMID- 9229190 TI - Lipid peroxidation in type 2 normolipidemic diabetic patients. AB - Vascular complications, as a consequence of atherosclerosis, are the main causes of morbidity and mortality in diabetes. Low density lipoprotein (LDL) oxidation is accepted as a relevant pathogenic mechanism in atherogenesis. The aim of this work was to study the relationship between lipid peroxidation (LPO) and metabolic control. LPO was evaluated in 40 type 2 normolipidemic diabetic patients by measuring thiobarbituric acid reactive substances (TBARS), in the plasma, using malondialdehyde (MDA), end product of the oxidation of polyunsaturated fatty acids, as a standard. Fast blood glucose (FBG), serum total cholesterol (TC) and serum triglycerides (TG) were evaluated by routine methods. Fructosamine (FR) was measured by the nitroblue tetrazolium (NBT) colorimetric test. An elevated level of lipid peroxides (P < 0.001) was observed in the plasma of diabetic patients (4.51 +/- 1.29 nmol/ml) as compared to normal subjects (3.54 +/- 1.00 nmol/ml). Lipid peroxides did not correlate with the FR levels, nor with FBG, TC and TG. These results show an increase of LPO in type 2 normolipidemic diabetic patients. Probably the mechanism for higher lipid peroxide levels in diabetes is multifactorial. Our study supports the hypothesis of a role of oxidative stress in diabetes independently of the lipid serum content. PMID- 9229191 TI - Improvement of the compliance with blood glucose monitoring in young insulin dependent diabetes mellitus patients by the Sensorlink system. AB - Recently, Medisense has introduced the Sensorlink system as a tool for retrieving the 125 last results of home blood glucose monitoring stored in patient's Pen 2 or Companion 2, unknown to them. Therefore we decided to check the compliance of type I diabetic adolescents and young adults with home blood glucose monitoring (HBGM) by comparing the blood glucose values noted in their log book and those retrieved by the Sensorlink and to evaluate an eventual subsequent effect both on compliance and glycated haemoglobin (HbA1c). The study was carried out in 60 insulin-dependent diabetes mellitus (IDDM) patients (33 women and 27 men) chosen according to two criteria: (1) the use of a Medisense Pen 2 or Companion 2; (2) autonomous self-monitoring of blood glucose, i.e. without parental supervision. They were aged 21.3 +/- 6.3 years with a diabetes duration of 11.6 +/- 7.0 years. HbA1c was measured by an HPLC method (N: 4.4-6.0%) before and after the first use of the Sensorlink and HBGM data of the log books were recorded. After the first use of the Sensorlink, the patients were warned of the retrieving data. The 60 patients were divided into two groups (same mean age and diabetes duration), according to the mean level of HbA1c before Sensorlink: < or = 7% (good control; n = 33); > 7% (insufficient control; n = 27). Cheating was unrelated to sex and occurred in 36 patients (60%; aged: 19.3 +/- 4.7 years), up to 100% in 13 of them (22%); five patients had no log book (8%; aged: 24.0 +/- 5.0 years); 19 patients (32%; aged 24.4 +/- 7.9 years) didn't cheat at all. After the use of the Sensorlink system, cheating dramatically decreased to zero. The effect of the Sensorlink system on improvement of HbA1c was statistically significant in the 27 patients with insufficient control before Sensorlink, since mean HbA1c level decreased from 8.0 +/- 0.9% to 7.5 +/- 1.1% (P < 0.05). In conclusion, non compliance with HBGM occurs in 2/3 of adolescents and young adults with IDDM. The Sensorlink system puts an end to this phenomenon and allows significant reduction of HbA1c levels in patients with insufficient metabolic control. PMID- 9229192 TI - Lipoprotein compositional abnormalities in type I diabetes: effect of improved glycaemic control. AB - Major lipoprotein mass and composition were assessed in 45 subjects with insulin dependent diabetes mellitus (IDDM), before and after 2 months of intensive insulin therapy (IIT) and in 40 healthy control subjects. As compared to the control group, diabetic subjects at baseline had higher low density lipoprotein (LDL) and lower high density lipoprotein (HDL) masses. Expressing each lipoprotein constituent as a percent of total lipoprotein mass, very low density lipoprotein (VLDL) of diabetic patients was enriched in cholesterol and phospholipid and depleted in triglyceride and protein; IDL had higher triglyceride and phospholipid and lower cholesterol and protein proportion; LDL was depleted in protein and enriched in triglyceride; HDL was depleted in protein and enriched in triglyceride, cholesterol and phospholipid. After 2 months of IIT, HbA1c fell from 10.3 +/- 2 to 7.5 +/- 2% (P < 0.0001) and so did VLDL mass, which was lower than in control subjects. In addition, LDL and HDL masses, as well as triglyceride and cholesterol proportion in IDL particles normalized. The other compositional abnormalities improved without complete normalization. Thus, intensive insulin therapy in IDDM subjects brought quantitative lipoprotein alterations to normal even subnormal range, while most of the composition abnormalities improved without reaching complete normalization. PMID- 9229193 TI - Accuracy, precision and user-acceptability of self blood glucose monitoring machines. AB - The performance of six self-monitoring blood glucose (SMBG) machines (Accutrend, Reflolux S, Companion 2, Glucometer GX, Glucometer IV and One Touch II) were examined using venous blood samples from 88 patients. Whole blood glucose (BG) values were measured by four machines from each brand. Machine-generated whole blood glucose (BG) values were corrected before comparison with laboratory plasma glucose values, measured by a glucose oxidase method. Based on error grid analysis, most of the corrected machine-generated BG values were clinically acceptable. Accutrend, Glucometer IV and Companion 2 showed the greatest consistency between machines of the same brand. Over 80% of corrected BG values generated by Glucometer IV fell within +/-10% of the reference values. One Touch II yielded the most reproducible results with a mean CV of 2.7% and was considered the most user friendly machine. More studies are required to examine the performance of these machines in the hands of patients. PMID- 9229194 TI - Very low calorie diet (VLCD): a useful alternative in the treatment of the obese NIDDM patient. AB - Conventional treatment of obese noninsulin dependent diabetes mellitus (NIDDM) patients is often unsatisfactory. In this study the efficacy of Modifast, a commercial very low calorie diet (VLCD), was evaluated in a population of obese poorly controlled NIDDM patients. The mechanisms of action of VLCD in these patients were also studied by comparing: (i) Plasma insulin and glucose profiles after a VLCD and an isocaloric mixed meal and (ii) plasma amino acid levels, both at baseline and after four weeks of VLCD treatment. A total of 14 obese NIDDM patients (M/F 7/7. median body mass index (BMI) 38.7 kg-2, interquartile range (IQ) 34.7-46.5 kg-2, waist circumference 116 cm, IQ 106-139 cm, insulin treated 7/14) with poor diabetic control (HbA1c 8.6%, IQ 7.8-10%) were studied. Patients were given a VLCD (425 kcal/day) for 12 weeks. At baseline, VLCD and isocaloric meal tests were performed on consecutive mornings. Fasting plasma amino acid levels were also determined at baseline and after 4 weeks of VLCD treatment. Weight, waist circumference, HbA1c, blood pressure, fasting plasma insulin, total cholesterol and triglyceride levels all fell significantly following VLCD treatment. Insulin therapy was able to be ceased in the seven insulin treated patients. Oral hypoglycaemic agent dosage fell from a median of eight (IQ 6-12) to two (IQ 0-8) tablets per day (P = 0.03) in patients initially on this form of therapy. Insulin secretion was higher after VLCD than isocaloric meal (P = 0.04). Fasting plasma alanine level fell from 512.0 (IQ 412.0-563.0) to 374.0 (IQ 342 472.0) mumol/l (P = 0.04) following VLCD treatment. In conclusion, the short term use of a VLCD is very effective in rapidly improving glycaemic control and promoting substantial weight loss in obese NIDDM patients. Moreover, a VLCD diet increases insulin secretion and reduces substrate for gluconeogenesis. Thus, VLCD treatment may improve glycaemic control by factors more than caloric restriction alone. PMID- 9229196 TI - Risk of noninsulin dependent diabetes mellitus conferred by obesity and central adiposity in different ethnic groups: a comparative analysis between Asian Indians, Mexican Americans and Whites. AB - Epidemiological data from Asian Indians from Madras (AI) and Mexican Americans (MA) and non-Hispanic Whites (NHW) from San Antonio heart study were compared to determine the possible contributions by the anthropometric measurements to the varied prevalence of noninsulin dependent diabetes mellitus (NIDDM) in these ethnic groups. MA had the highest rate of obesity (mean body mass index (BMI) 28.9 +/- 5.9 kg/m2) and the highest prevalence of diabetes (men 19.6%; women 11.8%, P < 0.001 vs other groups). NHW although had high rates of obesity (mean BMI 26.2 +/- 5.2 kg/m2) had low prevalence of diabetes (men 4.4%; women 5.7%) than the AI (men 9.9%; women 5.7%) (Mean BMI 22.3 +/- 4.4 kg/m2, P < 0.001). Although AI had lower BMI than MA, the risk conferred by BMI was similarly high in AI and MA and both the ethnic groups had higher risks than NHW. Impaired glucose tolerance (IGT) was also more prevalent in MA than in AI (men, MA vs AI, 11.8 vs 7.5%, P < 0.003; women 16.1 vs 5.5%, P < 0.001). NHW had lower prevalence of IGT in men (5.7%) and women (6.3%) which were significantly lower (P < 0.001) compared to MA only. Age and BMI were predictive factors of NIDDM in all, while waist to hip ratio (WHR) was significant only in AI and MA, although NHW had high WHR. This may be an indicator of differences in genetic susceptibility. This study also highlights the similarity in risk factors between AI and MA living in urban environment and the significance of distribution of adiposity in the comparatively lean AI. PMID- 9229195 TI - Acipimox attenuates hypertriglyceridemia in dyslipidemic noninsulin dependent diabetes mellitus patients without perturbation of insulin sensitivity and glycemic control. AB - Hyperlipidemia, hypertriglyceridemia in particular, is a common feature in patients with noninsulin dependent diabetes mellitus (NIDDM) and may associate with insulin insensitivity. Acipimox, being widely prescribed for treating hypertriglyceridemia, is also used in NIDDM patients for their dyslipidemia. In the present study, we evaluated the effect of acipimox in Chinese NIDDM patients with hypertriglyceridemia. A total of 16 patients enrolled in a double-blind, randomized, placebo-controlled and two-period crossover study. After an 8 week run-in period, patients were randomly assigned into two groups receiving either acipimox (250 mg, twice daily) or placebo treatment. A total of 12 weeks later, these two groups switched their treatment for an additional 12 weeks. Blood samples were collected at the end of the run-in period and then at 4-week intervals in the whole study for lipid profile. A modified insulin suppression test was performed at the ends of the run-in period, 12-week and 24-week treatment to assess changes in insulin sensitivity. Our results showed that acipimox significantly lowered serum total triglyceride while compared to those by placebo. However, no difference was observed in serum non-esterified fatty acid, low-density lipoprotein cholesterol, total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C) and HDL-C/ TC ratio between the two groups. Furthermore, glycemic indices and insulin sensitivity were similar during the base-line, placebo or acipimox periods. Taken together, our data suggest that acipimox significantly lowered TG without perturbation of insulin sensitivity in hypertriglyceridemic NIDDM patients. PMID- 9229197 TI - Urinary albumin excretion rate and cardiovascular disease in Spaniard type 2 diabetic patients. AB - To assess the prevalence of urinary albumin excretion abnormalities and their associations with cardiovascular disease or its classical risk factors in type 2 diabetes mellitus, 1348 clinic-proceeding patients have been studied retrospectively. The overnight urinary albumin excretion rate, blood pressure, smoking, ophthalmic and cardiovascular status, current therapies, estimates of glycemic control, plasma lipids, serum creatinine and uric acid have been ascertained. 767 (56.8%) patients were found normoalbuminuric, 461 (34.1%) microalbuminuric and 120 (8.9%) macroalbuminuric. In bivariate analyses, the urinary albumin excretion rate had statistically significant (P < 0.05) relationships with age, duration of diabetes, male sex, waist-to-hip ratio, systolic and diastolic pressure, coronary heart disease, cerebrovascular disease, peripheral vascular disease, hypertension, antihypertensive therapy, laser treated retinopathy, kind of treatment, smoking habit, fasting glycaemia, HbA1c, creatinine, uric acid, triglycerides, high density lipoprotein (HDL)-cholesterol and apolipoprotein B. Borderline statistically significant (P < 0.1) relationships were found with hypolipidaemic therapy, insulin dose, non-HDL cholesterol, apolipoprotein A1 and lipoprotein (a). In a multivariate stepwise logistic regression model, HbA1c, hypertension, male sex, age, diastolic blood pressure, coronary heart disease and body-mass index were sequentially selected as variables independently associated with microalbuminuria. Serum creatinine, HbA1c, male sex and hypertension were sequentially selected as independently associated with macroalbuminuria. Micro and macroalbuminuria are frequent abnormalities associated with poorly controlled and complicated disease, with overt cardiovascular disease and its classical risk factors as well as with the male sex. PMID- 9229198 TI - Interpreting recent trends in prostate cancer incidence and mortality. PMID- 9229199 TI - Benefits and risks of lowering serum cholesterol. PMID- 9229200 TI - Food labels, trans fatty acids, and coronary heart disease. PMID- 9229201 TI - Epidemiology and the internet. PMID- 9229202 TI - Prostate cancer incidence and mortality rates among white and black men. AB - This paper presents prostate cancer incidence and mortality rates in the United States by factors associated with the disease at diagnosis and explores racial differences in these rates. The analysis is based on population-based cancer data from the Surveillance, Epidemiology, and End Results (SEER) Program of the National Cancer Institute. Incidence rates increased sharply from 1973 to 1992 (more so among whites), with the increase attributed to screening and preferential finding of low-risk, slow-growing tumors (that is, length bias). Mortality rates increased slightly (more so among blacks). The 1988-1992 white and black prostate cancer mortality rates comprise cases diagnosed as early as 1973. These rates remained fairly constant for each stage category at diagnosis, except for black men diagnosed with localized or unstaged disease, in whom the rates increased. Of the white prostate cancer cases, 33% are diagnosed with localized disease, 16% are diagnosed with regional disease, and 38% are diagnosed with distant disease. The corresponding percentage for black patients are 31, 12, and 45%. Prostate cancer mortality rates for blacks diagnosed with localized, regional, distant, and unstaged disease are 1.9, 1.5, 2.4, and 2.0 times those of whites. The time from diagnosis to death depends on the stage and age at diagnosis, with the time somewhat longer for whites than blacks. This longer time period is due, in part, to better staging and length bias. The effect of therapy on prostate cancer mortality rates is unclear. PMID- 9229203 TI - Time trends in serum cholesterol before cancer death. AB - Following evidence of an association between low serum cholesterol and cancer from several prospective studies, this paper concentrates on individual time trends in serial measurements of serum cholesterol before a cancer-related death. The Framingham Heart Study contains repeated measurements of cholesterol at approximately 2-year intervals in 5,209 subjects, of whom 539 died from cancer during 30 years of follow-up. We quantify (1) the change in serum total cholesterol level before cancer death, and (2) the association between fall in serum total cholesterol and odds of cancer death. The mean fall in serum total cholesterol in the 4- to 6-year period before cancer death is 8.06 (95% confidence interval = 4.58-11.54) mg per dl, with some evidence of lowered cholesterol before that period. This pattern is corroborated by evidence of a substantially increased odds of cancer death if a large fall in cholesterol occurs over any 4- to 6-year period. We suggest that these time trends can plausibly be attributed to the effects of prevalent cancer on lowering serum cholesterol; our findings add weight to the argument that the low cholesterol cancer mortality relation does not arise because of any causal contribution of low serum total cholesterol to the risk of cancer. PMID- 9229204 TI - Decline in serum total cholesterol and the risk of death from cancer. AB - We investigated whether decline over time in serum cholesterol was associated with the risk of death from cancer in French men. We studied 6,230 working men, age 43-52 years in 1967-1972, who had at least three annual measurements of serum cholesterol. We estimated individual change over time in serum total cholesterol using within-person linear regression. During an average of 17 years of follow-up after the last examination, 747 subjects died from cancer. The multivariate adjusted relative risks for subjects in the fourth (highest increase in serum total cholesterol), third, and second quartiles, compared with men in the first quartile (who had a decrease in serum total cholesterol), were 0.70 [95% confidence interval (CI) = 0.56-0.87], 0.71 (95% CI = 0.57-0.88), and 0.74 (95% CI = 0.61-0.91), respectively. The group with the highest decline in cholesterol displayed an excess risk for most cancer sites. These associations were more pronounced in subjects whose weight remained stable or decreased over time than in those who gained weight. PMID- 9229205 TI - Margarine intake and subsequent coronary heart disease in men. AB - Margarine is a major source of trans fatty acids, the intake of which has risen since the early 20th century. Some data indicate that consumption of trans fatty acids increases the risk of coronary heart disease (CHD). In 1966-1969, 832 men from the Framingham Study, age 45-64 years and free of CHD, were administered a single 24-hour dietary recall, from which we estimated total daily margarine intake. We calculated CHD cumulative incidence rates and, using proportional hazards regression, CHD incidence rate ratios over 21 years of follow-up. Mean energy intake was 2,619 kcal; mean margarine intake was 1.8 (range 0-12) tsp per day. There were 267 incident cases of CHD. Age-adjusted CHD cumulative incidence rose over categories of margarine intake, but the increased risk was apparent only in the second half of the follow-up period. Adjusted for age and energy intake, the risk ratio for CHD for each increment of 1 teaspoon per day of margarine was 0.98 [95% confidence interval (CI) = 0.91-1.05] for the first 10 years of follow-up and 1.10 (95% CI = 1.04-1.17) for follow-up years 11-21. Adjustment for total fat intake and for cigarette smoking, glucose intolerance, left ventricular hypertrophy, body mass index, blood pressure, physical activity, and alcohol intake did not materially change the results. Butter intake did not predict CHD incidence. These data offer modest support to the hypothesis that margarine intake increases the risk of coronary heart disease. PMID- 9229206 TI - The relation of alcohol intake to coronary heart disease and all-cause mortality in a beer-drinking population. AB - Epidemiologic studies indicate that light to moderate alcohol consumption from beer, wine, or spirits is associated with a reduction in all-cause mortality, owing primarily to a reduced risk of coronary heart disease (CHD). To find out whether this protective effect of small to moderate amounts of alcohol could be confirmed in Germany, where much of the alcohol consumed is taken in the form of beer, we studied the relation between alcohol and CHD and total mortality in a population of southern Germany. We conducted a prospective cohort study from 1984 to 1992 among 1,071 men and 1,013 women, age 45-64 years at baseline, from the Augsburg region. Eighty-seven per cent of men and 56% of women reported drinking alcohol at baseline. Among drinkers, men had an average alcohol intake of 42 gm per day, of which 33 gm per day came from beer. Women who drank had an average alcohol intake of 16 gm per day and derived about half of it from beer and the other half from wine. During the 8 years of follow-up, 96 deaths (all causes) and 62 incident CHD events (nonfatal and fatal) occurred in men, and 45 deaths (all causes) occurred in women. Adjusting for a number of potential confounders, in men the adjusted hazard rate ratio (HRR) of CHD events for drinkers as compared with nondrinkers was 0.51 [95% confidence interval (CI) = 0.27-0.95]; this protective effect starts with the 0.1-19.9 gm per day alcohol category and does not change much with higher intake. In men, the adjusted total mortality HRR for drinkers as compared with nondrinkers was 0.59 (95% CI = 0.36-0.97). The total mortality HRRs for the different alcohol groups compared with nondrinkers show a U-shaped curve, with the lowest HRR of 0.46 (95% CI = 0.20-0.80) for the 20-39.9 gm per day alcohol group and an HRR of 1.04 (95% CI = 0.54-2.00) for the > or = 80 gm per day alcohol group. In women, the total mortality HRR for those drinking up to 19.9 gm per day as compared with nondrinkers was 0.46 (95% CI = 0.22-0.96). PMID- 9229207 TI - Does periconceptional multivitamin use reduce the risk for limb deficiency in offspring? AB - There is accumulating evidence that periconceptional multivitamin use may prevent the occurrence of some birth defects other than neural tube defects. Using data from the population-based Atlanta Birth Defects Case-Control Study, we investigated the possible association between periconceptional multivitamin use and the occurrence of limb deficiency. We examined the periconceptional use of multivitamins among mothers of 117 babies with nonsyndromic limb deficiency who were liveborn or stillborn to residents of metropolitan Atlanta from 1968 to 1980 and among mothers of 3,029 control babies born without birth defects who were randomly selected through birth certificates. We found that children whose mothers were periconceptional multivitamin users had a lower risk of having a limb deficiency [odds ratio (OR) = 0.47; 95% confidence interval (CI) = 0.23 0.97]. This protective effect, however, was mostly seen for transverse limb deficiency (OR = 0.30; 95% CI = 0.07-1.32) and not for longitudinal deficiency (including preaxial and postaxial deficiencies; OR = 1.03; 95% CI = 0.17-4.30). Adjustment for potential confounding factors did not change these findings. We found a trend of decreasing risk for all transverse limb deficiencies with earlier vitamin use. These data indicate that mothers' periconceptional multivitamin use may reduce the risk for some types of limb deficiency among their offspring. In addition, because we did not find the protective effect for all types of limb deficiency, the data may also indicate causal heterogeneity of limb deficiencies. PMID- 9229208 TI - Association between ambient carbon monoxide levels and hospitalizations for congestive heart failure in the elderly in 10 Canadian cities. AB - We examined the role that ambient air pollution plays in exacerbating cardiac disease by relating daily fluctuations in admissions to 134 hospitals for congestive heart failure in the elderly to daily variations in ambient concentrations of carbon monoxide, nitrogen dioxide, sulfur dioxide, ozone, and the coefficient of haze in Canada's 10 largest cities for the 11-year period 1981 1991 inclusive. We adjusted the hospitalization time series for seasonal, subseasonal, and weekly cycles and for hospital usage patterns. The logarithm of the daily high-hour ambient carbon monoxide concentration recorded on the day of admission displayed the strongest and most consistent association with hospitalization rates among the pollutants, after stratifying the time series by month of year and adjusting simultaneously for temperature, dew point, and the other ambient air pollutants. The relative risk for a change from 1 ppm to 3 ppm, the 25th and 75th percentiles of the exposure distribution, was 1.065 (95% confidence interval = 1.028-1.104). The regression coefficients of the other air pollutants were much more sensitive to simultaneous adjustment for either multiple pollutant or weather model specifications. PMID- 9229209 TI - Association among health habits, risk factors, and all-cause mortality in a black California population. AB - We evaluated dietary and other risk factors in a black California cohort. Baseline data were gathered in 1974 and 1976, and mortality follow-up continued through 1985. A study census questionnaire was returned from 3,299 subjects who lived in a household containing at least one Seventh-Day Adventist. Of these, 1,668 subjects also completed a detailed life-style and dietary questionnaire in 1976. Vital status was ascertained using church records and the California State death tapes. Mortality hazard ratios (HR; both sexes combined) across three increasing consumption levels were determined for nuts (1.00, 0.60, 0.56), fruits (1.00, 0.38, 0.57), and green salads (1.00, 0.54, 0.65). Consumption of meats appeared more hazardous for women, although there was no dose-response relation. Education (HR = 1.00, no college; 0.74, some college; 0.42, college graduate), male gender (HR = 1.55), diabetes mellitus (HR = 1.77), and hypertension (HR = 2.52) were independently associated with mortality, as was obesity, which had a curvilinear association in women and a linear association in men. Exercise was not associated with mortality after excluding those with morbidity at baseline. In summary, traditional risk factors operated with similar force in this black population. In addition, the frequent consumption of nuts, fruits, and green salads appears protective. PMID- 9229210 TI - Risk factors for sudden coronary death in the United States. AB - We assessed risk factors for sudden coronary death among persons without a history of coronary heart disease (unexpected sudden coronary death) and persons with a history of coronary heart disease (sudden coronary heart disease death). We analyzed national data to calculate death rates and odds ratios for both types of sudden coronary death. Among modifiable factors that we examined, only cigarette smoking increased risk for unexpected sudden coronary death [odds ratio (OR) = 1.8; 95% confidence interval (CI) = 1.2-2.7]. Diabetes mellitus (OR = 3.8; 95% CI = 2.5-5.8 for women), cigarette smoking (OR = 1.5; 95% CI = 1.0-2.1), and hypertension (OR = 1.4; 95% CI = 1.1-1.9) increased the risk for sudden coronary heart disease death. Etiologic factors for sudden death appear to differ depending on the presence or absence of coronary disease. With preexisting coronary disease, factors associated with chronic coronary disease may elevate sudden death risk; without coronary disease, factors that provoke ventricular arrhythmias may trigger sudden death. PMID- 9229211 TI - Prenatal and perinatal risk factors for breast cancer in young women. AB - There is increasing interest in the role of early life exposures in breast carcinogenesis, especially estrogen exposure in utero. Estrogen levels during pregnancy may be higher in twin pregnancies and among older women and slightly lower among smokers. We analyzed early life risk factors in a population-based case-control study in the United States of 2,202 breast cancer cases and 2,009 controls under age 55 years. Twins were at an increased risk of breast cancer compared with singletons (relative risk = 1.62; 95% confidence interval = 1.0 2.7), particularly women with a twin brother (relative risk = 2.06), a finding consistent with the observation of high estrogen levels in dizygotic twin pregnancies. Little association was seen between maternal age at birth and breast cancer risk. We carried out further analyses for 534 cases and 497 controls under age 45 years, using data from a questionnaire completed by their mothers relating to the daughters' early life exposures. There was no evidence of an effect of smoking or diethylstilbestrol exposure during pregnancy on daughters' breast cancer risk. A reduced breast cancer risk was seen among women who had been breastfed (relative risk = 0.74; 95% confidence interval = 0.6-1.0). These findings indicate some effect of early life exposures on breast cancer risk, although the role of estrogen exposure may be less central than previously suggested. PMID- 9229212 TI - Breastfeeding, menopause, and epithelial ovarian cancer. AB - No previous study has examined the modifying effect of menopausal status on the association between lactation and ovarian cancer risk. We recruited 824 epithelial ovarian cancer cases and 855 community controls in three Australian states, collecting reproductive and lactation histories by means of a contraceptive calendar and pregnancy and breastfeeding record. We report results in women with at least one liveborn infant for unsupplemented breastfeeding, in line with a biological model linking suppression of ovulation to reduction in ovarian cancer risk. We derived odds ratios from multiple logistic regression models including number of liveborn children, age, age at first or last birth, and other potential confounders, overall and by menopausal status. Estimates of relative risk of ovarian cancer per month of full lactation were 0.99 [95% confidence interval (CI) = 0.97-1.00] overall and 1.00 (95% CI = 0.99-1.01) and 0.98 (95% CI = 0.95-1.01) among post- and premenopausal women, respectively. We tailored a lactation variable to the incessant ovulation hypothesis by progressively discounting breastfeeding the longer after birth it occurred, finding odds ratios similar to those for the unmodified duration variable. We found no association of note among postmenopausal women. Breastfeeding seems to be somewhat protective against ovarian cancer, but only before menopause. PMID- 9229213 TI - Bone mass and subsequent risk of hip fracture. AB - We examined prospectively the associations of bone density and bone dimensions with risk of hip fracture using data from the first National Health and Nutrition Examination Survey and its three follow-up studies. A cohort of 1,489 white women age 45 years or older who received detailed medical examinations in the baseline survey in 1971-1975 were subsequently contacted in 1982-1984, 1986, and 1987. Bone density and hand bone dimensions at several sites were measured at baseline. Fifty incident hip fractures were identified during the follow-up studies. Using Cox regression analyses, we found a relative risk of 11 for women with bone density below the 5th percentile, compared with those above the 75th percentile (95% confidence interval = 2.2-58). Women with smaller external bone dimensions also faced increased risk of hip fracture (relative risk = 4.6 for dimensions below the 5th percentile vs above the 75th percentile; 95% confidence interval = 1.5-14). On the other hand, internal bone dimensions were not associated materially with hip fracture. PMID- 9229214 TI - Reliability and comparability of three dietary assessment methods for estimating fruit and vegetable intakes. AB - Although fruits and vegetables have been evaluated in numerous epidemiologic studies, few validation studies have examined fruits and vegetables. We examined the reproducibility and comparability of fruit and vegetable intakes estimated by diet records, food frequency questionnaires, and modules (brief food frequency questionnaires) in 101 control participants of a 1-year diet intervention trial. For each method, mean intakes at baseline and 3 months were generally within 0.3 serving per day for juice, fruits, vegetables, and total fruits and vegetables. In addition, Pearson correlations for the two time periods generally exceeded 0.55 for these four groups for each method. We evaluated comparability of intakes for 15 days of diet records, 1-year food frequency questionnaires, and modules, respectively. Mean total fruit and vegetable intakes were 6.3, 6.5, and 3.8 servings per day for diet records, food frequency questionnaires, and modules. For each pair-wise combination of methods, Pearson correlations exceeded 0.45 for juice, fruits, and total fruits and vegetables; correlations were lower for vegetables. Exact agreement in quintile assignment was less than 45%, however. These results indicate that estimates of fruit and vegetable intakes and disease associations may differ depending on the method used to assess fruit and vegetable intake. PMID- 9229215 TI - Educational achievement recorded on certificates of death compared with self report. AB - This study reports the agreement of educational achievement obtained from death certificates with self-report among 249 persons. Education was categorized as: less than high school, high school education, and greater than high school education. The overall agreement rate was 84%. Death certificates provided a slight underestimation of educational achievement. The sensitivity and specificity of the death certificate in classifying a decedent as having less than a high school education were both 90%. Educational achievement determined from death certificates has substantial validity, and any misclassification that may occur is likely to be similar across ethnicity and gender groups. PMID- 9229216 TI - The effect of measurement error on the determination of Helicobacter pylori prevalence. AB - We assessed the effect of serum-based antibody tests on the epidemiology of Helicobacter pylori infection. We took crude population prevalences of H. Pylori infection from existing publications in which antibody-based tests were used to determine prevalence. We then calculated 95% confidence intervals that included terms for study size, sensitivity, specificity, and, where possible, the sample size used to determine the validity of the antibody test. Specificity had a greater effect than sensitivity on the overestimation of most population-based estimates of H. pylori prevalence. The attributes of some antibody-based tests imply that no matter how large the study size, an accurate estimate of prevalence could not have been obtained. PMID- 9229217 TI - The alternative to epidemiologic theory: whatever works. PMID- 9229218 TI - The need for epidemiologic theory. PMID- 9229219 TI - Relative attributable risks. PMID- 9229220 TI - The need for exposure grouping strategies in studies of magnetic fields and childhood leukemia. PMID- 9229222 TI - The equilateral Lexis diagram. PMID- 9229221 TI - Confounding by indication and past clinical trials. PMID- 9229223 TI - Smoking and risk of hospitalization. PMID- 9229224 TI - Glycated low-density lipoprotein and atherogenesis: the missing link between diabetes mellitus and hypercholesterolaemia? PMID- 9229225 TI - Structural and compositional modifications of diabetic low-density lipoproteins influence their receptor-mediated uptake by hepatocytes. AB - Dyslipoproteinaemia is an important risk factor for the development of atherosclerosis in noninsulin-dependent diabetes mellitus (NIDDM). This study shows that the uptake of low-density lipoproteins (LDLs) prepared from the plasma of patients with NIDDM in cultured human hepatoma cells is largely reduced. In addition, diabetic LDL was less effective in suppressing intracellular cholesterol synthesis. This is because of physicochemical and biochemical differences between lipoproteins from diabetic and from normal individuals. LDL from patients with NIDDM was abnormal with regard to charge, the degree of glycation, the lipid composition and the conformation of the apolipoprotein B receptor-binding domain. The diminished receptor-mediated uptake of apolipoprotein B-containing lipoproteins in diabetic individuals most probably leads to the accumulation of these lipoproteins in vivo and may be of great importance to the pathogenesis of atheroclerosis in these patients. PMID- 9229226 TI - Expression of follicle-stimulating hormone receptor in human ovary. AB - The gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), are key hormones in the regulation of ovarian function. The acquisition of FSH receptors during folliculogenesis is believed to be a key event in the subsequent development of the follicle. However, the binding and biochemical properties of the human FSH receptor are not well-characterized owing to the low abundance of these receptors and the limited availability of human tissue. The binding experiments show that, while the affinity of the FSH receptor does not change through the menstrual cycle, the total number of FSH receptors in the leading follicles increases by about two-fold at mid-follicular phase compared with those at other periods. Northern blot analysis was used to measure relative levels of FSH receptor mRNA, and in situ hybridization was used to localize FSH receptor transcripts. Northern blot analysis of human ovaries detected two transcripts (4.1 and 2.4 kb) for the FSH receptor. The FSH receptor mRNA was abundant in the preovulatory follicle, with expression decreased by about 50% in the corpus luteum. Using in situ hybridization, FSH receptor mRNA was found to be confined to the granulosa cells of developing follicles. A reverse transcription polymerase chain reaction amplification was used to detect the expression of different isoforms of the FSH receptor mRNA in human corpus luteum and placenta. PMID- 9229227 TI - The effects of insulin on transport and metabolism of glucose in skeletal muscle from hyperthyroid and hypothyroid rats. AB - The effects of insulin on the rates of glucose disposal were studied in soleus muscles isolated from hyper- or hypothyroid rats. Treatment with triiodothyronine for 5 or 10 days decreased the sensitivity of glycogen synthesis but increased the sensitivity of lactate formation to insulin. The sensitivity of 3-O methylglucose to insulin was increased only after 10 days of treatment and was accompanied by an increase in the sensitivity of 2-deoxyglucose phosphorylation; however, 2-deoxyglucose and glucose 6-phosphate in response to insulin remained unaltered. In hypothyroidism, insulin-stimulated rates of 3-O-methylglucose transport and 2-deoxyglucose phosphorylation were decreased; however, at basal levels of insulin, 3-O-methylglucose transport was increased, while 2 deoxyglucose phosphorylation was normal. In these muscles, the sensitivity of lactate formation to insulin was decreased; this defect was improved after incubation of the muscles with prostaglandin E2. The results suggest: (a) in hyperthyroidism, insulin-stimulated rates of glucose utilization in muscle to form lactate are increased mainly because of a decrease in glycogen synthesis; when hyperthyroidism progresses in severity, increases in the sensitivity of glucose transport to insulin and in the activity of hexokinase may also be involved; (b) in hypothyroidism, the decrease in insulin-stimulated rates of glucose utilization is caused by decreased rates of glycolysis; (c) prostaglandins may be involved in the changes in sensitivity of glucose utilization to insulin observed in muscle in altered thyroid states. PMID- 9229228 TI - Antioxidant status in patients with uncomplicated insulin-dependent and non insulin-dependent diabetes mellitus. AB - Oxidative damage by free radicals has been implicated in the pathogenesis of vascular disease in diabetes. We compared the radical-scavenging antioxidant activity of serum from 28 patients with insulin-dependent diabetes mellitus and 24 patients with non-insulin-dependent diabetes mellitus uncomplicated by vascular disease with age-matched non-diabetic control subjects. Patients with insulin-dependent diabetes had significantly reduced total antioxidant activity (320.2 +/- 11.3 vs. 427.5 +/- 19.2 mumol L-1; P < 0.001). This was attributable to lower urate (209.4 +/- 10.4 vs. 297.1 +/- 16.7 mumol L-1; P < 0.001) and vitamin C levels (63.6 +/- 6.0 vs. 87.5 +/- 4.9 mumol L-1; P < 0.01). Patients with non-insulin-dependent diabetes had lower total antioxidant activity than age matched control subjects (433.8 +/- 25.4 vs. 473.9 +/- 30.2 mumol L-1; NS), reflecting lower urate (299.5 +/- 19.4 vs. 324.8 +/- 21.4 mumol L-1; NS) and vitamin C levels (38.6 +/- 5.7 vs. 58.5 +/- 5.3 mumol L-1; P < 0.05). Multiple regression analysis showed that urate, vitamin C and vitamin E were the major contributors to serum total antioxidant activity. These results show that diabetic patients have significant defects of antioxidant protection, which may increase vulnerability to oxidative damage and the development of diabetic complications. PMID- 9229229 TI - Bile acid inhibition of interferon activity in human lymphocytes: no evidence of oxidative stress. AB - Cholestasis and bile acids are two factors involved in resistance to interferon therapy in patients with chronic hepatitis C. As bile acids inhibit the biological activity of this cytokine in vitro and are capable of generating oxidative stress in hepatocytes, we investigated the potential involvement of such a mechanism in human lymphocytes. Thus, we evaluated (a) the effects of bile acids (0-200 mumol L-1) on lymphocyte reduced glutathione content and malondialdehyde production and (b) the ability of antioxidants to prevent the inhibitory effect of chenodeoxycholic acid on interferon-induced lymphocyte 2',5' oligoadenylate synthetase activity, an index of the biological activity of interferon. We found that treatment of lymphocytes with bile acids for 24 h did not induce malondialdehyde release or significantly modify cellular reduced glutathione content. Synthetic precursors of glutathione (N-acetylcysteine and S adenosylmethionine) and antioxidants (superoxide dismutase and catalase) had no preventive influence on the inhibitory effect of chenodeoxycholic acid on interferon-induced 2',5'-oligoadenylate synthetase activity. These negative results do not provide evidence for the use of glutathione precursors in cholestatic conditions associated with viral diseases. PMID- 9229230 TI - Relationship between lipoprotein(a) phenotypes and albumin excretion rate in non insulin-dependent diabetes mellitus: protective effect of 'null' phenotype? AB - The possible association between lipoprotein(a) [Lp(a)] and albumin excretion rate (AER) is a topic that generates conflicting views. In addition, Lp(a) phenotypes have not previously been considered as factors influencing AER. In order to clarify this issue, we studied 70 non-insulin-dependent diabetes mellitus (NIDDM) patients without clinically detectable macroangiopathy, 27 with microalbuminuria and 43 without it. Both groups were matched for the known variables that could influence AER and serum Lp(a) levels. Lp(a) was determined by enzyme-linked immunosorbent assay (ELISA), and Lp(a) phenotypes were assessed by electrophoresis followed by immunoblotting. Lp(a) phenotypes were grouped as follows: 'small' (F, S1 and S2), 'big' (S3 and S4) and 'null'. The NIDDM patients with microalbuminuria presented higher serum Lp(a) concentrations than the patients without it [15.7 mg dL-1 (95% CI 0.5-36.5) vs. 4.5 mg dL-1 (95% CI 0.1 18.5); P < 0.001] and a direct correlation between Lp(a) and AER was observed (r = 0.34; P < 0.01). AER was significantly different when Lp(a) phenotypes were considered ['small': median 19 micrograms min-1 (range 1-195); 'big': median 9.5 micrograms min-1 (range 1-186); 'null': 4 micrograms min-1 (range 1-9); P = 0.04]. None of the NIDDM patients with a 'null' phenotype showed an AER of > 10 micrograms min-1. In conclusion, this case-control study provides evidence that microalbuminuria is associated with high serum Lp(a) in NIDDM without clinically detectable macroangiopathy. Furthermore, NIDDM patients with a 'null' phenotype could be considered at low risk for the development of microalbuminuria. PMID- 9229231 TI - No acute effects of smoking and nicotine nasal spray on lipolysis measured by subcutaneous microdialysis. AB - Smoking is associated with insulin resistance, dyslipidaemia and markers of the insulin resistance syndrome. This study investigated adipose tissue lipolysis in situ by subcutaneous microdialysis twice in 10 healthy, male smokers after smoking four cigarettes over 2 h and after the administration of an equal amount of nicotine given as nasal spray (NNS). Glucose and insulin levels, in situ lipolysis and adipose tissue blood flow were studied in the post-absorptive state and after a 75-g oral glucose tolerance test (OGTT). Post-absorptively, acute smoking and NNS increased neither subcutaneous adipose tissue glycerol production nor plasma free fatty acid (FFA) or glycerol levels. After the OGTT, plasma insulin and lactate levels were significantly higher after smoking, whereas FFA levels were higher after NNS. Normal smoking or the administration of a normal dose of NNS caused only minor metabolic changes. Thus, it does not seem likely that increased lipolysis is an important contributor to the dyslipidaemia seen in smokers. PMID- 9229232 TI - Study of biochemical substrate and role of metalloproteinases in fascia transversalis from hernial processes. AB - The aim of this study was to examine the fascia transversalis (FT) from patients with direct and indirect hernia in an attempt to identify possible differences between each type of hernia. FT samples were obtained from 36 patients presenting inguinal hernia (23 indirect hernia and 13 direct hernia) who underwent surgery. We have analysed the ultrastructure of the fascia surrounding the hernial lesions, the proline and lysine hydroxylation in the tissue, the type I-type III collagen ratio and the presence of metalloproteinases. We have not detected ultrastructural differences in the collagen fibrils from FT in direct and indirect hernias. However, the interfibrillar matrix was more abundant in direct hernias, showing abundant electron-dense particles. No differences in proline hydroxylation were observed between each type of hernia. A small decrease in lysine hydroxylation was detected in patients with direct hernia. Enzyme-linked immunosorbent assays (ELISAs) showed no statistically significant differences in the type I-type III collagen absorbance ratios. Immunohistochemistry revealed no differences in the expression of matrix metalloproteinase-1. FT from patients presenting direct hernia showed a very strong staining vs. metalloproteinase-2 when compared with that observed in indirect hernia. PMID- 9229233 TI - Growth hormone improves cardiac function in rats with experimental myocardial infarction. AB - Accumulating evidence suggests from experimental and clinical studies beneficial effects of growth hormone (GH) on contractility, although concomitant cardiac hypertrophy, generally considered to be a cardiovascular risk factor, has also been reported. In the present study, we combine a rat model with impaired cardiac performance after myocardial infarction (MI) with echocardiographic evaluation of GH effects on cardiac structure and function. We have used a rat model with ligation of the left coronary artery in normal, growing male rats resulting in subsequent impaired cardiac performance. After 6 weeks' recovery, blind transthoracic echocardiography was performed to determine infarction size, cardiac geometry and performance. Rats with no signs of myocardial infarction were excluded from the study. After randomization, the rats were treated with daily s.c. injections of saline (n = 8) or recombinant human growth hormone (rhGH) (n = 6) at a dose of approximately 1 mg kg-1 body weight for 1 week. A new blind echocardiography examination was performed after treatment demonstrating a 13% increase in ejection fraction (EF) and a 50% increase in cardiac index in GH treated rats compared with control rats (P < 0.01). Moreover, GH caused a significant decrease in end-systolic volume. There were no significant changes in left ventricular (LV) or interventricular wall thickness, LV dimensions, heart rate or diastolic function. No effects were seen on LV weight, cardiac insulin like growth factor (IGF) I, IGF-I receptor and GH receptor mRNA content. GH in a physiological dose improves systolic function in an experimental model of heart failure without signs of hypertrophy, suggesting a potential role as a therapeutic agent in the treatment of heart failure and merits further investigation. PMID- 9229234 TI - Plasma endothelin-1 levels during transient acute myocardial ischaemia in men: effects of coronary revascularization. AB - The endothelium-derived peptide endothelin-1 (ET-1) was evaluated in 14 male patients [mean age 52.74 years (SEM 1.10)] affected by coronary artery disease during a bicycle electrocardiographic stress test and dipyridamole echocardiogram. Both tests were performed before and after coronary revascularization. Fourteen healthy male subjects served as controls [mean age 53.21 years (SEM 1.63)]. Baseline plasma endothelin-1 levels were higher (P < 0.0001) in ischaemic patients [1.81 pg mL-1 (0.15, n = 14)] than in control subjects [0.61 pg mL-1 (0.03, n = 14)], but did not increase with exercise in both groups. Similar results were obtained with dipyridamole infusion. Endothelin 1 levels significantly decreased after coronary revascularization [before: mean 1.81 pg mL-1 (SEM 0.15, n = 14); after: mean 1.16 pg mL-1 (SEM 0.11), P < 0.002], without changes in the peptide response to both tests. In conclusion, elevated plasma endothelin-1 concentrations were found in patients with stable angina compared with non-ischaemic subjects. No changes were observed during exercise or dipyridamole infusion in both groups. Coronary revascularization was followed by a significant decrease in plasma endothelin-1 levels. PMID- 9229235 TI - Glucagon-like peptide-1 (GLP-1): a trial of treatment in non-insulin-dependent diabetes mellitus. AB - Glucagon-like peptide-1 (7-36) amide (GLP-1) is released from the gut into the circulation after meals and is the most potent physiological insulinotropic hormone in man. In contrast to presently available therapeutic agents for non insulin-dependent diabetes mellitus (NIDDM), GLP-1 has the advantages of both suppressing glucagon secretion and delaying gastric emptying. We report the first chronic study of subcutaneous (s/c) GLP-1 treatment in NIDDM. Five patients with poorly controlled NIDDM were entered into a six-week, double-blind crossover trial. Each received three weeks treatment with s/c GLP-1 40 nmol or saline, given three times a day immediately before meals. A standardized test meal was given at the beginning and end of each treatment period. GLP-1 reduced plasma glucose area under the curve (AUC) following the standard test meal by 25% (AUC, 0-180 mins, GLP-1 start of treatment 482.2 +/- 38.2 vs. saline start of treatment 635.7 +/- 45.4 mmol min L-1, F = 16.4, P < 0.02). The beneficial effect of GLP-1 on plasma glucose concentration was fully maintained for the three-week treatment period. Plasma glucagon levels were significantly lower for 60 min postprandially after GLP-1 treatment. In this group of patients there was no significant increase in postprandial insulin levels with GLP-1. We have demonstrated a significant improvement in postprandial glycaemic control with s/c GLP-1 treatment that was fully maintained over a three-week treatment period. GLP-1 improves glycaemic control even in the absence of an insulinotropic effect and is a potential treatment for NIDDM. PMID- 9229236 TI - Experimental hydatid disease of the liver. AB - Hydatid disease caused by Echinococcus granulosis is an endemic problem in many parts of the world and the liver is the most frequently involved organ. A suitable experimental model for hydatid disease of the liver, resembling naturally infected livers, is established in this study by injection of protoscolices via the mesenteric vein of the mice. This model can be used in assessment of the efficacy of various agents in treatment of hydatid disease of the liver. PMID- 9229237 TI - Neural regulation of viscera elicited by individual interpretations of life experiences. PMID- 9229238 TI - Autonomic nervous control of heart rate orienting and alpha responses in dog. AB - The object of this experiment is to study the autonomic nervous control of alpha responses elicited in classical conditioning. Twenty-two mongrel dogs were trained in classical discriminative conditioning. Typical two-phase tachycardic responses were observed in positive (CS+) trials while only the earliest, phase 1, response was observed in negative (CS-) trials. The phase 1 response, which was identical in CS+ and CS-trials, was compared in dogs before and after selective SA-nodal parasympathectomy (N = 7) and beta-adrenergic blockade (N = 11). The phase 1 tachycardic response was eliminated by selective SA-nodal parasympathectomy but not by beta-adrenergic blockade. We conclude that the phase 1 response observed in both CS+ and CS-trials with similar time sequence and magnitude is an alpha response. The heart rate orienting response results from a withdrawal of parasympathetic activity with little or no change in sympathetic tone. PMID- 9229239 TI - Evidence of the origin of specific spontaneous head turns during intertrial intervals. AB - Direction and the frequency of spontaneous head movements during the ITIs following forward and backward paired trials were compared to an acquisition of a conditioned orienting (alpha) response directed to the side of the tone source. The head movements were analyzed from video recordings using classification of head turns to preferred and to nonpreferred directions. The results showed a significant increase in the alpha responses during the forward paired conditioning to the preferred direction and rapid extinction during the subsequent backward conditioning sessions. Spontaneous head movements during the ITIs increased to the same preferred direction as the conditioned alpha responses. The results of this experiment suggest that the response initially elicited by the CS can later appear as "spontaneous," instrumental behavior, the form and the nature of which is determined by the characteristics of the conditioned alpha response developing as a result of classical conditioning. PMID- 9229240 TI - Is there conscious choice in directed mutation, phenocopies, and related phenomena? An answer based on quantum measurement theory. AB - In a previous article (Goswami, 1997), it was suggested that an application of quantum measurement theory under the auspices of a monistic idealist ontology (that consciousness is the ground of being) can solve many difficult problems of neo-Darwinism, e.g., alternating rapid creativity and homeostasis observed in evolution and the directionality, origin, and nature of life. In this article, we propose an epigenetic quantum mechanism to explain the connection of developmental processes and evolution, as has been evidenced in such controversial phenomena as directed mutation and phenocopies. PMID- 9229241 TI - On the dynamics of dying. AB - This study is concerned with dynamic processes that underly the rapid, degenerative changes associated with the "dying" stage of the multicellular organism's life cycle. The interaction between negative and positive feedback cycles is discussed: negative feedback cycles underly the superstability characteristic of health and illness. When negative feedback cycles fade in the dying phase of life, positive feedback cycles, previously held in check by the negative feedback cycles to which they had been coupled, rise explosively, driving physiologic variables from their normal values towards extremes. This results in the rapid downturn that we associate with dying--an accelerating disintegration terminating in death. A medical case history is analyzed. PMID- 9229242 TI - Pavlov and the mind-body problem. AB - I. P. Pavlov claimed that the mind-body problem would ultimately be resolved by empirical methods, rather than by rational arguments. A committed monist, Pavlov was confronted by dualism in the case of an hysterical person. Under normal conditions, her body's left side was insensitive to pain, but when she was hypnotized, there was a reversal of her sensitivity to pain, with the right side becoming insensitive. Pavlov acknowledged that the divergence between stimulation and response suggested dualism, yet condemned his disciple G.P. Zelenyi as well as Charles S. Sherrington, for their dualistic tendencies. Pavlov's continuous adherence to monism it attributed to the influence of popular scientific books that he read during his adolescence. The books maintained that science was based upon monism. Pavlov proposed that by introducing the concept of emotions, an hysterical person's condition could be explained within the framework of his theory of higher nervous activity, thereby obviating the need to change his paradigm. PMID- 9229244 TI - Stanley Joe Edmonds (1909-1995). PMID- 9229243 TI - Neurally mediated hypotension, chronic fatigue syndrome and upper aerodigestive tract reflexes. PMID- 9229245 TI - Cytochromes in the respiratory chain of helminth mitochondria. AB - Parasitic helminths exhibit greater diversity in energy metabolism than do the host animals and many have exploited unique respiratory chains as adaptations to their natural habitats. Cytochromes are involved, not only in intracellular aerobic respiration found in free-living stages, but also in the reduction of relatively oxidized compounds such as fumarate during the adult stages of parasitic helminths. In addition, most helminths retain a significant capacity to produce energy via aerobic pathways and have a mammalian type respiratory chain in their mitochondria during their development in the host. In this review, we focus on recent advances in the study of cytochromes in the respiratory chain of parasitic helminths. These include the identification of unique features of anaerobic respiration in adult parasites, the elucidation of molecular structures of the components involved and an understanding of the developmental changes that occur during the life-cycle of these parasites. PMID- 9229246 TI - In vitro hatched oncospheres of Asian Taenia from Korea and Taiwan develop into cysticerci in the peritoneal cavity of female scid (severe combined immunodeficiency) mice. AB - In vitro hatched (but not activated) oncospheres of Asian Taenia obtained in Korea and Taiwan, prepared by the sodium hypochlorite method, rinsed with sterile PBS several times and adjusted to 5 x 10(4)/0.5 ml PBS, were injected intraperitoneally or subcutaneously into male or female scid mice of 3 different strains. When these scid mice were sacrificed 4 months later, the females harboured fully developed cysticerci either in the peritoneal cavity or under the back skin, whereas males did not. All cysticerci from the peritoneal cavity were easily recovered by rinsing the abdomen with PBS. Although most cysticerci recovered from pig liver usually become calcified within 1-2 months, in female scid mice they all increased in size and were viable. Intraperitoneal (i.p.) injection of in vitro hatched oncospheres is recommended for easier recovery of Asian Taenia metacestodes in laboratory animals. PMID- 9229247 TI - Differentiation and ultrastructure of the paruterine organs and paruterine capsules, in the nematotaeniid cestode Nematotaenia dispar (Goeze, 1782) Luhe, 1910, a parasite of amphibians. AB - Three types of egg-protecting envelopes of parenchymatic, uterine and embryonic origin have been distinguished in the cyclophyllidean cestode Nematotaenia dispar (Goeze, 1782) Luhe, 1910, a type species for the genus Nematotaenia and the family Nematotaeniidae. The present paper deals with the parenchymatic envelopes, which originate from the modified medullary parenchyma and are represented in this species by the paruterine organs and paruterine capsules. In pregravid proglottids they are composed of clongated myocytons, myofibrils and membranous anucleate cellular processes, containing a large amount of lipid droplets and some calcareous corpuscle cells. These cellular elements (CE) are separated from each other by abundant extracellular matrix (ECM), which consists primarily of an electron lucent ground substance with fine filaments embedded in it. The paruterine capsules of gravid proglottids are surrounded from the outside by a typical medullary parenchyma and are lined by a layer of the connective tissue. The paruterine organs and paruterine capsules show similar ultrastructure. During their histogenesis, all cellular elements undergo extensive flattening, followed by cellular deterioration, with simultaneous reduction in CE/ECM ratio. In the late gravid segments, paruterine capsule walls are very thick and consist of membranous sheets with large amounts of lipid droplets, which cause the cytoplasmic sheets to bulge. Ultrastructure of various types of parenchymatic envelopes in representatives of different cyclophyllidean families, such as paruterine organs in Nematotaeniidae and Mesocestoididae, uterine and parenchymatic egg capsules in Anoplocephalidae (Linstowiinae and Inermicapsiferinae, respectively), is compared. PMID- 9229248 TI - Comparison between patterns of pinworm infection (Aspiculuris tetraptera) in wild and laboratory strains of mice, Mus musculus. AB - Sixteen laboratory and 7 wild-derived strains of mice were infected with the pinworm Aspiculuris tetraptera in order to compare their resistance levels estimated by the intestinal parasite loads. It appears that (i) in 4 strains out of 23, females and males harbour different parasite loads; (ii) wild and laboratory mice display a broad range of infection levels when compared independently; (iii) the laboratory strains are more resistant than the wild ones. We suggest that (i) compared to sex, the strain (i.e. genetic) effect is the main parameter which determines the levels of infection; (ii) resistance was selected in laboratory strains during their breeding because of the parasite pressure present in captivity. PMID- 9229249 TI - Genetic and immunological adaptation of Heligmosomoides polygyrus in mice. AB - Two lines of Heligmosomoides polygyrus, QN and QA, were selected by passage, respectively, through naive and immune Quackenbush (Q) mice and their biology and capacity to induce immune responses in homologous Q and heterologous low- (SL) and high-responder (RH) mice were assessed. QA H. polygyrus survived the impact of otherwise protective immunity in Q mice better than did QN parasites, especially after a secondary infection. The enhanced survival of QA parasites was observed also in SL but not in RH mice. Infections with QA phenotypes induced a reduced antibody response and lower eosinophilia compared to QN parasites in Q mice. The total numbers of nucleated cells in the mesenteric lymph node (MLN) and spleen increased to different extents according to the genotype of the mice and the phenotype of parasite used: the increase was most profound in RH, least in SL and intermediate in Q mice; QN induced more lymphocytosis in the MLN and spleen than did QA parasite phenotypes. B cells from MLN and spleen, stained with fluorochrome-conjugated antibody against mouse Ig, showed increased intensity of fluorescence in the flow cytometric assay after infection with H. polygyrus, but to different degrees: the intensity increased most in RH and least in SL mice; more in Q mice infected with QN than with QA H. polygyrus. These results suggest that adaptation of parasites to immunity of the host is associated with reduction of their immunogenicity and is specific to the immune status and genotype of the host to which the parasites have become adapted. PMID- 9229250 TI - The effect of infection with the abomasal nematode, Haemonchus contortus, on the avoidance behaviour of sheep in a motivational-choice test. AB - The behaviour of immune and non-immune sheep infected with H. contortus and undergoing a variety of experimental treatments was investigated in a motivational-choice test, the arena test. This test evaluates motivational state in sheep by pitting the motivation of test animals to approach a small flock of sheep against the motivation to avoid a human decoy located directly in front of the small flock. Approach distance is decreased by infection in immune ewes but was unaffected by infection in non-immune weaner lambs in the present study. Experimental drug-treatments with the opiate-antagonist nalorphine, the antihistamine chlorpheniramine, and the immunosuppressive glucocorticoid dexamethasone, affected avoidance behaviour but did not shed light on possible mechanisms involved in the changes observed when immune sheep are infected with the parasite. These substances may affect motivational state directly and not through a pharmacological action on processes triggered by infection in immune sheep. Arena tests conducted in immune ewes at 4, 7 and 11 days after challenge infection showed a fluctuating locomotor behaviour, which may arise from either the dynamics of a standard secondary immune response or particular antigens released during larval development. The immune-mediated changes in behaviour in the arena test will have entailed information-processing or cognitive pathways, but it is not known whether they also involved the physiological manifestations of emotion. PMID- 9229251 TI - Nutritional and respiratory pathways to parasitism exemplified in the Turbellaria. AB - Symbiosis is a dominant trait in the Platyhelminthes. The Neodermata (Aspidogastrea, Monogenea, Digenea, Udonellidea, Cestoda) are wholly parasitic and even the predominantly free-living Turbellaria have almost 200 species from 35 families living in permanent associations with other animals. In the simplest turbellarian symbioses, ectosymbiotes such as the Temnocephalida, some other Rhabdocoela and a few Tricladida live on the body surfaces or in the branchial chambers of their mainly arthropodan or chelonian hosts. They feed on the same types of prey as their free-living relatives but supplement their diet by opportunistic commensalism. Their digestive physiology and food reserves are the same as in free-living species. The entosymbiotic Umagillidae, Graffillidae, Pterastericolidae, Fecamplidae and Acholadidae live in internal body cavities or body wall derivatives of echinoderms, molluscs or arthropods and show increasing metabolic dependence on their hosts. Patterns of digestive physiology and food storage generally differ markedly from those of ectosymbiotic and free-living species. Some umagillids, in echinoids, feed as entozoic predators on co symbiotic protozoa, supplemented by opportunistic ingestion of the hosts' ingesta, gut cells or coelomocytes. Others, in holothurians, feed mainly on gut cells, which also provide some digestive enzymes, and to a lesser extent on host ingesta and co-symbiotes. Graffillids, in molluscs, lack endogenous digestive enzymes and rely entirely on those taken in with host ingesta and gut tissues. Pterastericolids, in asteroids, similarly utilise gut tissues both as food and enzyme sources. The climax to metabolic dependence occurs in the Fecamplidae and Acholadidae. The former, in crustacean haemocoels and myzostomid tissues, lack conventional alimentary systems and absorb soluble nutrients through the epidermis. In the latter the only known species lives in the tube feet of its asteroid host, lacks a normal endodermal gut, but has a modified epidermis performing both digestive and absorptive functions. Most of these entosymbiotes show a shift from the lipid storage characteristic of free-living and ectosymbiotic species to the glycogen storage predominating in the Neodermata. In both groups this emphasis on carbohydrate metabolism is often independent of the PO2 of their environment. Both groups also show high fecundity and it is suggested that there is a direct relationship between this and glycogen storage. High fecundity, while clearly of adaptive value in entosymbiotes, is arguably primarily related to the assured food supply conferred by the entosymbiotic habit and thus can be viewed as a consequence of the latter rather than a prerequisite for it. Some entosymbiotic Turbellaria have evolved physiologically active haemoglobins, allowing them to abstract oxygen preferentially from host tissues; some have also evolved facultative glycolytic mechanisms comparable to those of the Cestoda. All these adaptations to ecto- and entosymbiotic life in the Turbellaria exemplify possible pathways to wholly parasitic lifestyles, with total metabolic dependence on the hosts, which may have been followed during the evolution of the Neodermata. PMID- 9229252 TI - Nutritional adaptations to parasitism within the platyhelminthes. AB - Some of the most significant alterations to the basic turbellarian plan are evident in the adaptations that relate to the acquisition of food by parasitic flatworms, reflecting the most potent of selection pressures in initiating and maintaining the host-parasite association. Nutritionally, ectoparasitic monogeneans show most correspondence with the predatory turbellarians, with certain monopisthocotylean members feeding by means of a protrusible pharynx and extracorporeal digestion, as skin-browsers of fish, with extensive intracellular digestion involving lysosomal enzymes in a well-differentiated gut. The more sheltered vascularised gill chamber of fish provides many polyopisthocotylean monogeneans with a totally renewable and more comprehensive diet in the form of blood, but haematophagy has necessitated a number of digestive adaptations, not least in resolving the problem of intracellular accumulations of haematin pigment. Haematophagy is the predominant feeding strategy of digeneans, but in contrast to monogeneans digestion of blood is largely extracellular; in schistosomes digestion is rapid, involving a battery of cathepsin-like cysteine proteinases and aminopeptidases. The external surfaces of all parasitic flatworms depart from turbellarian character and are composed of a multifunctional syncytial tegument, which is permeable to a variety of small organic solutes, some crossing by passive diffusion, others via facilitated or active mediated transport. The relative roles of the tegument and gut in trematode nutrition are difficult to assess, but can be related to the nature of the microhabitat within the host. Cestodes are highly adapted intestinal parasites bereft of any vestige of gut, and their tegument has become elaborated into a sophisticated and highly efficient digestive-absorptive layer, rivalling the vertebrate mucosa in its ability to gain kinetic advantage in the selective uptake of nutrient at the host parasite interface. The patterns of energy metabolism in adult flatworm parasites are generally anaerobic and based on glycogen, with abbreviated metabolic pathways and the loss of biosynthetic capacities. PMID- 9229253 TI - Reproduction and host-location among the parasitic platyhelminthes. AB - This review examines briefly the reproductive capacity of representatives of the 4 principal groups of platyhelminths, the "Turbellaria", Monogenea, Digenea and Cestoda. Of the flatworms, 3 main groups are wholly parasitic (monogeneans; digeneans; cestodes). Among the largely free-living "Turbellaria", there are several parasitic representatives in some families (Umagillidae; Graffillidae; Pterastericolidae; Fecampiidae; Acholadidae). Endoparasitic platyhelminths with complex life-cycles produce large numbers of eggs and numbers of offspring are increased further in the digeneans and a few cestodes by asexual multiplication. Like their free-living relatives, most ectosymbiotic and ento- and ectoparasitic flatworms ("turbellarians" and monogeneans) produce, on the whole, far fewer eggs and progeny but are still successful organisms in terms of their numbers of species and diversity. Estimates of parasite fecundity from in vivo experiments are needed for representatives from all flatworm groups. For those parasites that are host-specific, the particular species of host provides a predictable target to be located. Adaptations displayed by the eggs and infective stages of some flatworms increase their chances of finding and recognising their specific host and these are reviewed: attachment of eggs to their "host"; egg hatching in response to host chemicals; rhythmical emergence; special behaviours of infective stages; host recognition. PMID- 9229254 TI - Origin of the epidermis in parasitic platyhelminths. AB - The epidermis of members of the major parasitic taxon Neodermata is distinctive among flatworms, being a syncytial, insunk, non-ciliated epidermis that develops through a wholesale replacement of larval epidermis at metamorphosis when the larva attacks a host. How it arose in evolution from what must have been a turbellarian-like ancestor is not immediately evident. While many turbellarian flatworms have also adopted a symbiotic way of life, the literature on ultrastructure of epidermis in these symbionts shows quite a variety of morphologies, many not so different from that of their free-living relatives. Various turbellarians do have syncytial or insunk epidermises or reduction of epidermal ciliation as is characteristic of the Neodermata, but co-occurrence in a single turbellarian of all features common to neodermatans has not been reported. Urastoma cyprinae, for example, which is ectosymbiotic on bivalves, has a ciliated cellular epidermis that is little different from what is known of epidermises of its free-living relatives. The endoparasitic Anoplodium hymanae, from the coelom of sea cucumbers, also bears a ciliated cellular epidermis, as is typical of many other rhabdocoels, but it shows marked phagocytic activity as well as incorporation of endosymbiotic bacteria. The closest similarity to neodermatan epidermis is that of the turbellarian Genostoma kozloffi, an ectosymbiont of the crustacean Nebalia: covering the bulk of the body is a non ciliated syncytium with multiple branching connections to insunk nucleated portions, much as in epidermis of adult neodermatans and, on its ventral surface, is a field of ciliated cellular insunk epidermis resembling the epidermis of some larval neodermatans. Developmental clues to the origin of the neodermatan epidermis can be seen in turbellarian embryos. Before hatching, embryos of proseriate and triclad embryos go through 3 generations of epidermis, each replacing the next; 2 generations of epidermis are reported in the literature on rhabdocoel embryos. This process of replacement parallels the epidermal replacement that larval neodermatans undergo at metamorphosis. Ultrastructural study of developing acoel, polyclad and macrostomid embryos shows that they, too, have epidermal replacement and growth through immigration of deeper-lying cells, comparable to the processes seen in higher flatworms. Succession of distinct generations of epidermis in such animals as the proseriates, triclads and rhabdocoels is probably an adaptation to development of ectolecithal eggs, providing the means for the embryo to use yolk that resides in vitellocytes, outside its blastomeres. We propose that the Neodermata has taken advantage of this developmental mechanism, producing successive generations of epidermal cells even in its larval stages, to counter the defenses of hosts. PMID- 9229255 TI - The origins of parasitism in the platyhelminthes: a summary interpreted on the basis of recent literature. AB - A summary is given of the 4 contributions on the origins of parasitism in the Platyhelminthes. Recent ecological literature is used to interpret some of the findings. In particular, recent findings on rugged fitness landscapes, the increasing difficulty of long-jump adaptations, complexity catastrophes, and empty phenotypic space are discussed in order to find an explanation for the large number of Neodermata and the scarcity of parasitic turbellarians; it is concluded that evolutionary stasis of symbiotic turbellarians that are "trapped" in their particular niches is responsible for the small number of symbiotic species of turbellarians, rather than competitive exclusion by the numerous neodermatans. The earliest neodermatans were probably already dependent on the production of many offspring; hence the protoneodermatan was probably a species preadapted to parasitism by high fecundity. Monogenea produce few offspring in spite of their rich and secure food supply; the protomonogenean was either a species with a complex behaviour pattern for habitat selection or, alternatively, a species with high fecundity subsequently reduced to permit the evolution of more complex behaviour patterns (switch from r- to K-strategy). The finding that embryonic replacement of the epidermis is shared by several turbellarian groups, a developmental pattern possibly used and modified in the formation of the neodermis in the Neodermata, can be interpreted as supporting the views that Neodermata and turbellarians with epidermal replacement are all monophyletic, or alternatively, that a similar developmental pattern has arisen several times due to similar selection pressures. However, the possibility of horizontal character transfer should also be considered for explaining similar characters, including similar developmental patterns, in apparently not closely related groups. Horizontal gene transfer and principles for demonstrating horizontal character transfer are discussed. PMID- 9229256 TI - Variables related to differences in standardized test outcomes for children with autism. AB - The purpose of this experiment was to assess whether manipulation of variables related to motivation and attention in children with autism would influence performance on standardized tests. Two different testing conditions were compared: One consisted of the usual standardized testing procedures; during the other, specific variables that were hypothesized to relate to each child's responsiveness to task stimuli were manipulated. Data were collected in the context of a repeated reversals experimental design with condition order varied within and across children. Six children participated in a total of 44 separate testing sessions, controlled for order of conditions, number of sessions, and type of test. Results showed consistent differences between the two conditions, suggesting that improving motivation and attention in children with autism may considerably influence test performance and interpretation. Findings are discussed in relation to the difficulty in administering and interpreting changes in performance on standardized tests with this population. PMID- 9229258 TI - Social perception in children with autism: an attentional deficit? AB - Research suggests that the attentional deficits found in children with autism may be related to impairments in social functioning (e.g., Courchesne et al., 1994a, 1994b; Lewy & Dawson, 1992; Schreibman & Lovaas, 1973). In the present investigation, 14 children with autism, 14 mentally handicapped, and 14 typically functioning children participated in a study designed to investigate the effects of number of social cues on the ability to interpret social situations. Participants were shown videotaped vignettes of child-child interactions in which the number of cues leading to the correct interpretation of the story varied from one to four (i.e., tone, content, nonverbal, or nonverbal with object). Subjects were then asked a series of questions which varied in degree of complexity. Overall, results indicated that children with autism performed as well as both groups of comparison subjects on general attention questions (i.e., identification of number and gender of interactants) and social perception questions relating to stories containing one cue. However, children with autism performed more poorly than both comparison groups on social perception questions relating to stories containing multiple cues. Results are discussed in terms of an attentional dysfunction hypothesis of autism. PMID- 9229259 TI - Sensory-perceptual abnormalities in autism: a case for more research? AB - Sensory-perceptual abnormalities in people with autism are discussed from two perspectives: published firsthand accounts and existing psychological research evidence. A range of abnormalities, including hyper- and hyposensitivity, sensory distortion and overload, and multichannel receptivity and processing difficulties, are described in firsthand accounts and frequently portrayed as central to the autistic experience. A number of dangers are inherent in uncritically accepting these accounts at face value and in any wider generalization to the autistic population as a whole. Evidence from clinical studies suggests that unusual sensory responses are present in a majority of autistic children, that they are manifested very early in development, and that they may be linked with other aspects of autistic behavior. In addition, experimental studies using a range of indices have found evidence of unusual responses to sensory stimuli in autistic subjects. However the clinical and experimental research to date suffers from serious methodological limitations and more systematic investigation is warranted. Key issues for future psychological research in the area are identified. PMID- 9229257 TI - A comparison of speaking and mute individuals with autism and autistic-like conditions on the Autism Behavior Checklist. AB - The item, total, and subscale scores on the Autism Behavior Checklist (ABC) were compared for 155 mute and 335 speaking individuals with autism spectrum disorders. Although no significant difference was observed between the groups on the ABC total score, the mute group demonstrated significantly more pathology on 21 of 57 items and 3 of 5 subscales. The speaking group obtained significantly higher scores on only 8 items and 1 subscale (Language). The appropriateness of providing greater pathology scores on expressive language items to speaking, rather than to mute, individuals is called into question. The authors speculate whether the expressive language items are weighted too heavily, in regard both to the Language subscale and to the ABC total score. If the expressive language items were removed, the mute group would have significantly higher ABC total scores and therefore a greater degree of autism severity. PMID- 9229260 TI - Autistic children's responses to separation and reunion with their mothers. AB - Observed 16 autistic, 16 normal, and 16 Down syndrome children (age 3-6 years during separation and reunion with their mother in a laboratory playroom over three sessions. Children's responses to separation and reunion were assigned to one of five behavioral patterns that were weighted for intensity or level of response. No differences were found between groups in their behavioral responses during separation or reunion. Moreover, children in each group altered their responses according to the environmental setting which was varied over the three sessions. However, the autistic and Down syndrome groups did differ from the normal group in their consistency of behavioral patterns over the three observation sessions; both the former groups showed more individual variation in their separation and reunion patterns indicating that the expression of these patterns may be influenced by their associated developmental delay. PMID- 9229262 TI - Brief report: prompted pretend play in autism. PMID- 9229261 TI - Risperidone and explosive aggressive autism. AB - Many autistic patients with mental retardation have difficulties with explosivity and aggression. They often prove resistant to various pharmacotherapeutic interventions. In this study, 11 male outpatients (mean 18.3 years) were administered risperidone in an open-label fashion. The risperidone was started at 0.5 mg daily, and titrated upwards until maximum clinical benefit occurred. Serial clinical interviews were conducted, and Conners Parent-Teacher Questionnaires (short form) were completed by the caretakers. Substantial clinical improvement was noted almost immediately in each patient, with aggression, self-injury, explosivity, and poor sleep hygiene most improved. The modal dose for optimal response was 0.5 mg bid. Weight gain was a significant side effect (average velocity of 0.47 kg per week), while none of the patients experienced extrapyramidal side effects. PMID- 9229263 TI - Brief report: autistic children's attentiveness and responsivity improve after touch therapy. PMID- 9229264 TI - The method of stimulated serial repetitions of gymnastic exercises in therapy of autistic children. PMID- 9229265 TI - Side effects associated with psychoactive medication in individuals with autism. PMID- 9229266 TI - The TEACCH strategy in mentally retarded children with autism: a multidimensional assessment. Pilot study. Treatment and Education of Autistic and Communication Handicapped children. PMID- 9229267 TI - When is a significant change not significant? PMID- 9229268 TI - Impact of technology change and managed care on medical oncology. PMID- 9229269 TI - Strategies for ritual speeches: tips from a professor of speech. PMID- 9229270 TI - A comparison of the Dutch Blueprint standards (theory) with the experiences of students in clerkships in Groningen (practice). AB - BACKGROUND: A comparison was made between the Dutch national objectives for the education of medical doctors (in terms of clinical experiences and skills) which are stated in the Blueprint, and the extents to which the objectives were realized in six clerkships at the Faculty of Medical Sciences in Groningen. METHODS: From each clerkship, 40 complete logbooks were analyzed. RESULTS: On the average, the clerkships did not fully meet the national objectives, but they did offer clinical experiences and skills that are not mentioned in the formal objectives. Students varied significantly in their clinical experiences. CONCLUSIONS: The design of the clerkships should be improved to make them more concordant with national goals. This relates to both cancer education and medical education in general. PMID- 9229271 TI - An innovative curriculum plan for advanced practice in oncology nursing. AB - BACKGROUND: The Advanced Practice in Oncology Nursing Program was designed to prepare graduates to manage the cancer experience through the delivery of comprehensive, holistic, oncology-focused care to individuals, families, and communities in a variety of settings. METHODS: Theoretical course work and clinical practicum are required to complete the degree plan. Students are actively recruited from urban, rural, and underserved population settings. The program is committed to fostering a teaching-learning paradigm that facilitates self-directed learning. The program's basic tenet is that all course offerings will be designed in a distributed learning/distance learning method. Clinical experiences are accomplished in or near the student's home community. RESULTS: Development and implementation of the program are in process. Shaping a curriculum and learning environment to be consonant with a health care system in a constant state of reform flux is a challenging task. CONCLUSION: This program's long-term challenge is to remain flexible, collaborative, and futuristic while promoting the expansion of advanced practice in oncology nursing. PMID- 9229272 TI - Evaluation of oral cancer screening. AB - BACKGROUND: Oral cancer screening procedures are designed to collectively allow early detection of cancers in a body area accessible to visual and physical examination, as well as to facilitate timely treatment, awareness, and the ongoing education of the public. METHODS: A state fair was selected for this activity because of the availability of a random population compatible with meeting these goals. A total of 1,151 individuals participated in this free elective activity. RESULTS: Of this number 4.17% were deemed to have oral pathologic states necessitating professional intervention, and 1.82% were clinically diagnosed as having potential dysplastic or precancerous lesions. No clinical oral cancer was detected in this population. Nevertheless, by virtue of screening and detecting clinically premalignant lesions, the screening test advanced the diagnosis of potential oral cancers. CONCLUSIONS: The outcome adds support to oral cancer screening as a procedure applicable in reducing morbidity and mortality from oral cancers. PMID- 9229273 TI - Knowledge of, attitudes toward, and barriers to cancer control and screening among primary care physicians in Egypt: the need for postgraduate medical education. AB - METHODS: The authors surveyed 177 primary care physicians in Menofeia, in the Nile Delta area of Egypt, to test their knowledge of, attitudes toward, and perceived barriers to cancer control and screening. RESULTS: The physicians viewed cigarette smoking and radiation exposure as the most important cancer risk factors, followed by occupation, family history, and sun exposure. The majority of the physicians saw diet as contributing little or nothing to cancer risk. Most of the physicians lacked knowledge about early cancer detection and screening. Junior practitioners, in particular, reported a lack of information about liver cancer and hepatitis viruses despite the prevalence of these viruses in the country. Large proportions of mid-career and senior primary care physicians who had no postgraduate education cited lack of knowledge and not being familiar with an approach to cancer prevention as reasons for not performing screening activities. CONCLUSION: The importance of smoking, diet, and early detection of common cancer types should receive more attention in the undergraduate medical curriculum of Egypt. Health and medical education authorities in Egypt should authorize more postgraduate education opportunities for senior primary care physicians, aimed at increasing their knowledge of prevention and improving their attitudes toward primary cancer prevention and screening. PMID- 9229274 TI - The readability levels of cancer-prevention materials targeting African Americans. AB - BACKGROUND: Several studies have examined the readability levels of cancer prevention materials, but the readability levels of materials targeting African Americans have not been documented. The Cancer Prevention Materials and African Americans project, funded in 1994 by the Texas Cancer Council, was developed to assess the readability levels and cultural sensitivity of cancer-prevention materials targeting African Americans. METHODS: This study assessed the readability of 100 cancer-prevention materials using McLaughlin's SMOG grading. Illustrations and point sizes were also examined by research staff members. RESULTS: Seventy-two percent of the material contained illustrations and 88% were printed using a 12-point font. An overall mean SMOG grade of 9.32 was found, which suggests that ninth graders can read and comprehend the text. CONCLUSIONS: Although this grade level is lower than those indicated by previous studies, many of the printed materials may not be appropriate for African Americans at high risk for cancer: those with low incomes and little education. Health professionals should focus on decreasing the reading levels of print materials, pretesting audiences to determine their actual reading levels, and examining other factors that influence readability and comprehension. PMID- 9229275 TI - The reliability of recollections of family history: implications for the medical provider. AB - BACKGROUND: Identification by medical providers of families at higher risks for breast cancer relies on patients' recollection of family histories. While patients seem to report the occurrences of breast cancer in their first-degree relatives accurately, little is known about the precision of reports of ages at diagnosis of cancers in relatives, one of the most revealing clues with regard to hereditary transmission. Moreover, the methods used in previous studies to elicit the family reports render their applicability to the medical setting questionable. METHODS: Confirmatory pathology records were sought and compared with reports of breast cancer events among 125 first-degree relatives by 68 women with breast cancer and 37 women without the disease. RESULTS: Sixty-seven (90.5%) of the reports of the occurrence of breast cancer in relatives by affected women and 32 (97.0%) of those by unaffected women were accurate. Women reporting several affected relatives often overreported the presence of breast cancer events. Nearly 89% of the reports of age at diagnosis were correct within five years. The average error in the reports was 2.0 years. The precision of reports of age at diagnosis did not differ according to the educational level of the proband, the age at which the relative had been diagnosed, or the type of relative affected. However, the mean error in reporting the age at diagnosis of relatives was significantly larger among probands 70 years old or older compared with younger probands. CONCLUSIONS: These results suggest that relying on reports by patients with and without breast cancer should not critically affect the assessment of breast-cancer risks for family members. Nevertheless, verification by examination of pathology records is justified when decisions about patient management and referral for genetic counseling are made based on reports of several affected relatives. PMID- 9229276 TI - The effects of the PDQ patient information file (PIF) on patients' knowledge, enrollment in clinical trials, and satisfaction. AB - BACKGROUND: This study examined the outcomes of providing a copy of the PDQ Patient Information File (PIF) to cancer patients. METHODS: Patients with cervical, endometrial, and ovarian cancers were randomized to two groups: 1) verbal communication only and 2) verbal communication plus PIF. Cancer knowledge and satisfaction with the PIF and the information received were assessed with telephone interviews. Clinical trial registries were reviewed to determine enrollment in clinical trials. RESULTS: The overall reaction to the PIF was good or excellent for 92% of the patients surveyed, but there was no significant difference between the two groups in their cancer knowledge, enrollment in clinical trials, or satisfaction with the information they received from their physicians. The majority of patients from both groups lacked basic knowledge about their disease, did not use any source of information other than their physicians and/or nurses, and were satisfied with the information they received. CONCLUSIONS: Patients appreciate receiving written cancer information, although it may not increase their cancer knowledge. PMID- 9229277 TI - Patients' perspectives on the management of emotional distress in primary care settings. AB - OBJECTIVE: To investigate how important treatment for emotional distress is to primary care patients in general and to primary care patients with depression, and to evaluate the types of mental health interventions they desire. DESIGN: Patient surveys. SETTING: Five private primary care practices. MEASUREMENTS AND MAIN RESULTS: Patients' desire for treatment of emotional distress and for specific types of mental health interventions were measured, as well as patients' ratings of the impact of emotional distress, the frequency of depressive symptoms, and mental health functioning. Of the 403 patients, 33% felt that it was "somewhat important" and 30% thought it was "extremely important" that their physician tries to help them with their emotional distress. Patient desire for this help was significantly related to a diagnosis of depression (p < .001), perceptions about the impact of emotional distress (p < .001), and mental health functioning (p < .001). Among patients with presumptive diagnoses of major and minor depression, 84% and 79%, respectively, felt that it was at least somewhat important that they receive this help from their physician. Sixty-one percent of all primary care patients surveyed and 69% of depressed patients desired counseling: 23% of all patients and 33% of depressed patients wanted a medication: and 11% of all patients and 5% of depressed patients desired a referral to a mental health specialist. CONCLUSIONS: A majority of these primary care patients and almost all of the depressed patients felt that it was at least somewhat important to receive help from their physician for emotional distress. The desire for this help seems to be related to the severity of the mental health problem. Most of the patients wanted counseling, but relatively few desired a referral to a mental health specialist. PMID- 9229278 TI - Utilization of outpatient diagnostic imaging. Does the physician's gender play a role? AB - OBJECTIVE: To examine physician and patient characteristics related to the ordering of imaging studies in a general medicine practice and to determine whether physician gender influences ordering patterns. DESIGN: Retrospective cohort study. SETTING: Hospital-based academic general medicine practice of 29 attending physicians. PATIENTS: All 8,203 visits by 5,011 patients during a 6 month period. METHODS: For each visit the following variables were abstracted from the electronic patient record: patient age, patient gender, visit urgency, visit type, and physician seen. All diagnostic imaging studies performed within 30 days of each outpatient visit were identified from the hospital's Radiology Information System. Screening mammography was not included in the analysis. Physician variables included gender and years since medical school graduation. Logistic regression analysis was used to evaluate the effect of various patient, physician, and visit characteristics on the probability of a diagnostic imaging study being ordered. RESULTS: Patient age, urgent visits, visit frequency, and the gender of the physician were all significantly related to the ordering of an imaging study. Correcting for all other factors, the ordering of an imaging study during an outpatient medical visit was 40% more likely if the physician was female (odds ratio = 1.40; 95% confidence interval [CI] 1.01, 1.95). Female physicians were 62% more likely (95% CI 0.99, 2.64) than male physicians to order an imaging study for a male patient and 21% more likely (95% CI 0.87, 1.69) to order an imaging study for a female patient. CONCLUSIONS: Physician gender is a predictor of whether an outpatient medical visit generates an imaging study. Reasons for this observation are unclear, but may be the result of different practice styles of male and female physicians or unmeasured patient characteristics. PMID- 9229279 TI - Increasing the use of advance directives in medical outpatients. AB - OBJECTIVE: We studied whether a simple educational intervention would increase patient completion of advance directives and discussions on end-of-life issues. DESIGN: Randomized, controlled trial. SETTING: Outpatient clinic of a teaching hospital. SUBJECTS: One hundred eighty-seven outpatients of a primary care internal medicine clinic. INTERVENTION: Study subjects attended a 1-hour interactive seminar and received an informational pamphlet and advance directive forms. Control subjects received by mail the pamphlet and forms only. MEASUREMENTS AND MAIN RESULTS: Completion of the advance directive was the main measurement. There were no significant differences in baseline characteristics of either group. Follow-up at 1 month revealed advance directive completion in 38% of study versus 24% of control subjects (p = .04), and discussions on advance planning in 73% of study versus 57% of control subjects (p = .02). Patients most likely to complete the documents were white, married, or attendees at the educational seminar. CONCLUSIONS: Interactive group seminars for medical outpatients increased discussions and use of written advance directives. PMID- 9229280 TI - A regional evaluation of variation in low-severity hospital admissions. AB - OBJECTIVE: Determine patient and hospital-level variation in proportions of low severity admissions. DESIGN: Retrospective cohort study. SETTING: Thirty hospitals in a large metropolitan region. PATIENTS: A total of 43,209 consecutive eligible patients discharged in 1991 through 1993 with congestive heart failure (n = 25,213) or pneumonia (n = 17,995). MEASUREMENTS AND MAIN RESULTS: Admission severity of illness was measured from validated multivariable models that estimated the risk of in-hospital death; models were based on clinical data abstracted from patients' medical records. Admissions were categorized as "low severity" if the predicted risk of death was less than 1%. Nearly 15% of patients (n = 6,382) were categorized as low-severity admissions. Compared with other patients, low-severity admissions were more likely (p < .001) to be nonwhite and to have Medicaid or be uninsured. Low-severity admissions had shorter median length of stay (4 vs 7 days; p < .001), but accounted for 10% of the total number of hospital days. For congestive heart failure, proportions of low-severity admissions across hospitals ranged from 10% to 25%; 12 hospitals had rates that were significantly different (p < .01) than the overall rate of 17%. For pneumonia, proportions ranged from 3% to 22%; 12 hospitals had rates different from the overall rate of 12%. Variation across hospitals remained after adjusting for patient sociodemographic factors. CONCLUSIONS: Rates of low-severity admissions for congestive heart failure and pneumonia varied across hospitals and were higher among nonwhite and poorly insured patients. Although the current study does not identify causes of this variability, possible explanations include differences in access to ambulatory services, decisions to admit patients for clinical indications unrelated to the risk of hospital mortality, and variability in admission practices of individual physicians and hospitals. The development of protocols for ambulatory management of low-severity patients and improvement of access to outpatient care would most likely decrease the utilization of more costly hospital services. PMID- 9229281 TI - Measuring symptomatic and functional recovery in patients with community-acquired pneumonia. AB - OBJECTIVE: To determine the rates of resolution of symptoms and return to premorbid health status and assess the association of these outcomes with health care utilization in patients with community-acquired pneumonia. DESIGN: A prospective, multicenter cohort study. SETTING: Inpatient and outpatient facilities at three university hospitals, one community hospital, and one staff model health maintenance organization. PATIENTS: Five hundred seventy-six adults (aged > or = 18 years) with clinical and radiographic evidence of pneumonia, judged by a validated pneumonia severity index to be at low risk of dying. MEASUREMENTS AND MAIN RESULTS: The presence and severity of five symptoms (cough, fatigue, dyspnea, sputum, and chest pain) were recorded through questionnaires administered at four time points: 0, 7, 30, and 90 days from the time of radiographic diagnosis of pneumonia. A summary symptom score was tabulated as the sum of the five individual severity scores. Patients also provided responses to the Medical Outcomes Study 36-Item Short Form Health Survey (SF-36) and reported the number of and reason for outpatient physician visits. Symptoms and health status 30 days before pneumonia onset (prepneumonia) were obtained at the initial interview. All symptoms, except pleuritic chest pain, were still commonly reported at 30 days, and the prevalence of each symptom at 90 days was still nearly twice prepneumonia levels. Physical health measures derived from the SF-36 Form declined significantly at presentation but continued to improve over all three follow-up time periods. Patients with elevated symptom scores at day 7 or day 30 were significantly more likely to report pneumonia-related ambulatory care visits at the subsequent day 30 or day 90 interviews, respectively. CONCLUSIONS: Disease-specific symptom resolution and recovery of the premorbid physical health status requires more than 30 days for many patients with pneumonia. Delayed resolution of symptoms is associated with increased utilization of outpatient physician visits. PMID- 9229282 TI - Identification of patient attitudes and preferences regarding treatment of depression. AB - OBJECTIVES: To identify attitudes that influence patient help-seeking behavior and aspects of treatment that influence patient preferences for management of depression. DESIGN: Three focus group discussions (two patient groups stratified by race and one professional group). Questions addressed experience with depression, help-seeking behaviors, treatment preferences, and perceived barriers to mental health care. SETTING: Academic medical center. PATIENTS/PARTICIPANTS: Eight black patients and eight white patients with depression: seven health care professionals (four physicians and three social workers). MEASUREMENTS AND MAIN RESULTS: Discussions were audiotaped, transcribed, and reviewed independently by two investigators to identify and group distinct comments into categories with specific themes. Differences were adjudicated by a third investigator. Comments within categories were then checked for relevance and consistency by a health services researcher and a psychiatrist. More than 90% of the 806 comments could be grouped into one of 16 categories. Black patients raised more concerns than white patients regarding spirituality and stigma. Patients made more comments than professionals regarding the impact of spirituality, social support systems, coping strategies, life experiences, patient-provider relationships, and attributes of specific treatments. They discussed the role these factors played in their help-seeking behavior and adherence to treatment. CONCLUSIONS: In-depth focus group discussions with depressed patients can provide valuable and unique information about patient experiences and concerns regarding treatment for depression. Clinicians, researchers, and policymakers need to incorporate the range of factors identified by patients into their decision making for individuals with depression. PMID- 9229284 TI - Development and validation of a geriatrics knowledge test for primary care residents. AB - This study reports the development and preliminary validation of an instrument to measure geriatrics knowledge of primary care residents. A 23-item test was developed using questions selected from the American Geriatrics Society's Geriatrics Review Syllabus. Ninety-six internal medicine and family practice residents, 14 geriatrics fellows, and 11 geriatrics faculty members participated in the study. Findings support the reliability (Cronbach's alpha = 0.66) and validity (content and "known groups") of this short test. Predictive validity and sensitivity of the test to changes in knowledge will have to be further explored as residents progress through their training. PMID- 9229283 TI - Case-finding instruments for depression. Two questions are as good as many. AB - OBJECTIVE: To determine the validity of a two-question case-finding instrument for depression as compared with six previously validated instruments. DESIGN: The test characteristics of a two-question case-finding instrument that asks about depressed mood and anhedonia were compared with six common case-finding instruments, using the Quick Diagnostic Interview Schedule as a criterion standard for the diagnosis of major depression. SETTING: Urgent care clinic at the San Francisco Department of Veterans Affairs Medical Center. PARTICIPANTS: Five hundred thirty-six consecutive adult patients without mania or schizophrenia. MEASUREMENTS AND MAIN RESULTS: Measurements were two questions from the Primary Care Evaluation of Mental Disorders patient questionnaire, both the long and short forms of the Center for Epidemiologic Studies Depression Scale, both the long and short forms of the Book Depression Inventory, the Symptom-Driven Diagnostic System for Primary Care, the Medical Outcomes Study depression measure, and the Quick Diagnostic Interview Schedule. The prevalence of depression, as determined by the standardized interview, was 18% (97 of 536). Overall, the case-finding instruments had sensitivities of 89% to 96% and specificities of 51% to 72% for diagnosing major depression. A positive response to the two-item instrument had a sensitivity of 96% (95% confidence interval [CI], 90-99%) and a specificity of 57% (95% CI 53-62%). Areas under the receiver operating characteristic curves were similar for all of the instruments, with a range of 0.82 to 0.89. CONCLUSIONS: The two-question case-finding instrument is a useful measure for detecting depression in primary care. It has similar test characteristics to other case-finding instruments and is less time-consuming. PMID- 9229285 TI - Emotions and medicine. What do patients expect from their physicians? PMID- 9229286 TI - Conjunctival pallor and the presence of anemia. PMID- 9229288 TI - Histologically diagnosed Helicobacter pylori in heart transplant recipients. AB - BACKGROUND: The role of Helicobacter pylori in the pathogenesis of nonautoimmune gastritis and peptic ulceration is well recognized. H. pylori is widely prevalent in the general population, but the incidence among heart transplant recipients has not been reported. Furthermore, the natural history of this infection may be modified by immunosuppression. METHODS: Gastric and duodenal biopsy specimens from 47 heart transplant recipients were examined over a period of 44 months. RESULTS: Twenty-three (49%) patients had H. pylori infection (15 men, 8 women; mean age 49 [range 35 to 59] years). Eight of the 23 (35%) had symptoms. These eight patients were treated for H. pylori with bismuth, metronidazole, and amoxicillin, followed by maintenance H2-receptor antagonists. Dyspepsia continued in six of these patients, with persistence or recurrence of H. pylori being demonstrated in four. CONCLUSIONS: This study shows that although histologically diagnosed H. pylori infection is widely prevalent among heart transplant recipients, this prevalence is very similar to the general population. Immunosuppression may play a role in the recurrence or persistence of this infection and may diminish the mucosal inflammatory response to the organism. PMID- 9229287 TI - Pathologic findings of latissimus dorsi muscle graft in dynamic cardiomyoplasty: clinical implications. AB - BACKGROUND: We hypothesize that the integrity of the latissimus dorsi muscle graft used to wrap the heart may affect the clinical outcome of patients undergoing dynamic cardiomyoplasty. METHODS: By correlating the pathologic findings with their clinical course in five patients who died 1 month to 6 years after dynamic cardiomyoplasty operation, we sought to discern findings that might shed light on the pathophysiology of cardiomyoplasty. RESULTS: Of the two patients who had a limited clinical response, one had an atrophic, edematous latissimus dorsi muscle with fatty infiltration resulting from cardiac cachexia, and the other had insufficient length of latissimus dorsi muscle to cover a large heart. The remaining patients responded well clinically without signs of pump failure and died at various intervals, mostly of arrhythmias. Autopsy findings included the following: (1) one patient with ischemic cardiomyopathy as the underlying disease had development of rich vascularity in the interface between the muscle wrap and the epicardium; whereas in four others with idiopathic cardiomyopathy, such evidence of collateralization was far less evident. (2) There was a variation in the skeletal muscle transformation achieved, with the fraction type I fatigue-resistant fiber in the muscle wrap ranging from 60% to 100%, in spite of the identical transformation protocol used. Such variation is believed to be genetically based. (3) In one patient, the skeletal muscle was paced to contract at 30 to 50 times/minute (2:1 ratio) for more than 5 years. Nevertheless, the pathologic specimen of the muscle wrap showed only minimal interstitial fibrosis. (4) Relatively thin muscle wrap around the heart found at autopsy could be atrophy but most likely was related to muscle transformation, which is known to reduce muscle mass and increase capillary density. (5) All skeletal muscle grafts showed geometric conformation to the shape of the epicardium and grossly looked as if they were an additional layer of the ventricular wall. Such conformation may facilitate the modulation of the ventricular remodelling process in the failing heart, as has been described both in clinical and experimental studies. CONCLUSIONS: Our findings are consistent with and support a number of mechanisms proposed for cardiomyoplasty. Thus preservation of latissimus dorsi muscle graft integrity may be important in the success of dynamic cardiomyoplasty. PMID- 9229289 TI - Absence of endocardial lymphoid infiltrates (Quilty lesions) in nonheart transplant recipients treated with cyclosporine. AB - The clinical records and autopsy material from 14 patients treated with cyclosporine who were not recipients of cardiac allografts were reviewed to further study the nature of endocardial lymphoid infiltrates (quilty lesions). Although there was well-documented exposure to cyclosporine, no endocardial lymphoid infiltrates were identified in these cases, providing evidence that the endocardial-based Quilty lesion is not the result of the direct effect of cyclosporine and may represent a localized form of heart rejection confined to the endocardium. PMID- 9229290 TI - Symptom distress three months after heart transplantation. AB - BACKGROUND: Although symptoms of heart failure abate after heart transplantation, other symptoms caused by the surgery, immunosuppressant drugs, and complications can be new sources of symptom distress for patients after operation. METHODS: This two-site National Institutes of Health study compared symptom distress in 173 adult heart transplant recipients from before operation to 3 months after operation. The Heart Transplant Symptom Scale was used to measure 92 symptoms related to heart disease and heart failure, transplantation, medication side effects, and complications commonly found in this population. Analysis was via paired t tests with Bonferroni correction. Most patients (93%) were receiving a triple immunosuppressant regimen of cyclosporine, azathioprine, and prednisone. RESULTS: Total symptom distress decreased significantly (p = 0.013) from before operation to 3 months after heart transplantation. The 23 symptoms that decreased the most (p = 0.000) after operation accounted for a cumulative total reduction of 583% less symptom distress. These symptoms were primarily cardiopulmonary, neuromuscular, and emotional. The 10 symptoms that worsened the most (p = 0.000) after operation accounted for a cumulative total increase of 284% more symptom distress. These symptoms were primarily dermatologic, neurologic, and gastrointestinal and were all side effects of prednisone and cyclosporine. CONCLUSIONS: The net change in symptom distress resulted in 299% less symptom distress in this cohort at 3 months after heart transplantation. This significant improvement in symptom outcomes scientifically documents the effectiveness of heart transplantation in reducing symptoms of heart failure, along with accompanying emotional symptoms. These research findings therefore reinforce and support the positive symptom outcomes often reported anecdotally in clinical practice. PMID- 9229291 TI - Cyclosporine-induced hypertension: evidence for maintained baroreflex circulatory control. AB - BACKGROUND: The clinical use of cyclosporine as an immunosuppressive agent enhanced long-term survival in transplant recipients at the expense of a high incidence of induced hypertension. Altered neurovegetative (autonomic) cardiovascular control is suspected as a mechanism of this form of hypertension. METHODS: Spectral analysis of systolic arterial pressure and R-R interval variability (electrocardiographic recordings) were performed, and the index alpha of baroreflex gain was computed in four groups of subjects matched for age: 13 orthotopic heart transplant recipients; 13 solid organ transplant recipients; 13 patients with essential hypertension; and 18 control subjects with normal blood pressure. All but the control subjects were treated with similar dihydropyridine calcium entry blockers. Heart and solid organ transplant recipients also received cyclosporine. RESULTS: R-R variance was lowest in the heart transplant recipients. The spectral profile of R-R interval was suggestive of sympathetic predominance in the patients with hypertension, but not in the solid organ transplant recipients or the control subjects. Systolic blood pressure variability and low frequency component (a marker of sympathetic vasomotor modulation) were similar in the four groups. The index alpha was 1.8 +/- 2.2 in heart transplant recipients, 11.7 +/- 6.6 in solid organ transplant recipients, 7.3 +/- 3.6 in patients with hypertension, and 13.5 +/- 6.4 msec/mm Hg in control subjects (p = 0.0001). CONCLUSIONS: These data indicate that (1) cyclosporine induced hypertension in heart transplant recipients is associated with a loss of baroreflex function as a result of cardiac denervation-related uncoupling; (2) compared with patients with hypertension, organ transplant recipients with hypertension demonstrated a maintained baroreflex function as indicated by a lack of reduction of the index alpha; (3) baroreflex heart rate control in dihydropyridine-treated cyclosporine-induced hypertension is well maintained. PMID- 9229292 TI - Time-dependent decrease of presynaptic inotropic supersensitivity: physiological evidence of sympathetic reinnervation after heart transplantation. AB - BACKGROUND: Sympathetic cardiac denervation of the transplanted human heart causes a loss of the presynaptic neuronal uptake1-mechanism with consecutive supersensitivity to uptake1-dependent catecholamines. A return of neuronal function (reinnervation) should result in a decrease of supersensitivity to catecholamines subjected to this uptake system and thus may alter the inotropic regulation. METHODS: Inotropic dose-response curves were compared in 12 patients who were studied 3 to 15 months after transplantation (early) and 17 patients who were studied 23 to 156 months after transplantation (late) with isoproterenol (uptake1-independent) and epinephrine (uptake1-dependent). The inotropic response to increasing doses of isoproterenol (5 to 20 ng/kg per min) and epinephrine (10 to 40 ng/kg per min) was assessed with echocardiography as increase of the systolic pressure/dimension ratio (delta P/D) and of the rate-corrected velocity of circumferential fiber shortening (delta Vcfc). RESULTS: Inotropic dose/response curves to isoproterenol were identical in the early and late recipients (during 20 ng/kg per min. isoproterenol: delta P/D 2.07 +/- 1.36 vs 2.18 +/- 1.42 mm Hg/mm; delta Vcfc 1.55 +/- 0.33 vs 1.40 +/- 0.38 square root of min-1 x %/ms), indicating an unchanged inotropic effect mediated by the postsynaptic beta-receptor/effector system. However, the inotropic response to epinephrine in early recipients was significantly attenuated in the late recipients (during 40 ng/kg per min. epinephrine: delta P/D 3.35 +/- 2.06 vs 1.51 +/- 0.68 mm Hg/mm, p < 0.01; delta Vcfc 1.80 +/- 0.42 vs 1.05 +/- 0.35 square root of min-1 x %/ms, p < 0.001). CONCLUSIONS: These findings provide evidence for an at least partial restoration of the neuronal catecholamine uptake and are consistent with a time-dependent sympathetic reinnervation after heart transplantation. Restoration of neuronal uptake seems to be of functional importance, because it profoundly alters the inotropic effect of circulating endogenous catecholamines in long-term survivors after heart transplantation. PMID- 9229293 TI - Long-term left ventricular assist device support: a novel pump rate challenge exercise protocol to monitor native left ventricular function. AB - A novel, hemodynamically guided exercise protocol with two different left ventricular assist device settings in two long-term recipients is presented. This protocol allows for quantitation of the contribution of the native left ventricle to total cardiac output. It facilitates estimation of the risk associated with device dysfunction, as well as prediction of left ventricular recovery and the potential for weaning. PMID- 9229294 TI - The effect of reduced myocardial cyclic AMP content on the response to milrinone in the isolated guinea pig heart. AB - BACKGROUND: Cardiac beta receptor down-regulation is associated with a reduction of tissue cyclic adenosine monophosphate (AMP) content. Milrinone exerts its effects by inhibiting the metabolism of existing cyclic AMP. The purpose of this study was to evaluate the effect of reduced myocardial cyclic AMP content on the pharmacologic action of milrinone. METHODS: A reduction of myocardial cyclic AMP content was produced by creating catecholamine depletion in the hearts of adult guinea pigs with intraperitoneal reserpine. Control animals received the reserpine vehicle. Isolated heart perfusion was maintained with modified Krebs buffer, and hearts were paced at 270 beats/min. A latex balloon and transducer tipped catheter were inserted into the left ventricle. Isovolemic work was maintained at a constant balloon volume. Hearts from control and reserpine treated animals were perfused for 20 minutes with buffer containing either no milrinone, 1.7 x 10(-6), or 1.0 x 10(-4) mol/L milrinone (n = 12 for each dose). Maximal positive and negative dP/dt were assessed. The hearts were then frozen and cyclic AMP was measured. RESULTS: Cyclic AMP content was significantly lower in the reserpine-treated hearts at each milrinone concentration (0.33 +/- 0.01 vs 0.46 +/- 0.01; 0.33 +/- 0.01 vs 0.53 +/- 0.01; 0.30 +/- 0.01 vs 0.61 +/- 0.02 pmol/mg wet weight, p < 0.05). Milrinone significantly increased positive and negative dP/dtmax (p < 0.05), but no difference was observed between control and reserpine-treated hearts. CONCLUSIONS: Endogenous catecholamine depletion reduces myocardial cyclic AMP content but does not attenuate the response to milrinone in the isolated heart. PMID- 9229295 TI - Time course of coronary endothelial dysfunction in acute untreated rejection after heterotopic heart transplantation. AB - BACKGROUND: Endothelial dysfunction is one of the early events leading to atherosclerosis. It occurs early after orthotopic heart transplantation and precedes the appearance of accelerated graft coronary artery disease believed to stem from chronic rejection of the endothelium. Acute rejection may contribute to the development of graft vasculopathy. METHODS: To assess the time course and specific mechanisms of coronary endothelial dysfunction in acute untreated rejection, a swine model of retroperitoneal heterotopic heart transplantation was used. Large white swine (age 10 +/- 2 weeks, weight 25 +/- 5 kg) were serum-typed for class I antigen of the swine leukocyte antigen system and selected to ensure a similar degree of incompatibility. Donor hearts were preserved with normothermic blood cardioplegia and regional hypothermia; the mean ischemic time was 64 +/- 15 minutes. Myocardial contractility decreased from day 5 (normal) to day 14 (weak), but electrical activity was preserved. All coronary arteries were patent, and International Society for Heart and Lung Transplantation grade 4 rejection was present in all hearts beyond 5 days. The endothelial function of epicardial coronary arterial rings of native and transplanted hearts was studied in organ chambers filled with modified Krebs-Ringer bicarbonate solution and compared 1, 5, 9, and 14 days after transplantation. RESULTS: Maximal endothelium independent relaxations were unaffected at all stages. Endothelium-dependent relaxations to serotonin and alpha 2-adrenergic agonist UK 14304 (which activate receptors coupled to Gi-proteins) and to sodium fluoride (a direct G-protein activator) deteriorated progressively over time. At 14 days maximal relaxations to the calcium ionophore A23187, adenosine diphosphate, and bradykinin were also reduced, but to a lesser degree than those to serotonin and sodium fluoride. Histomorphometric studies of the allograft coronary artery rings showed progressive intimal hyperplasia from day 5 to day 14, with an increase in the incidence from 29% +/- 8.3% to 61.5% +/- 12%. CONCLUSIONS: These studies show that endothelial dysfunction in untreated acute rejection after heart transplantation develops beyond 5 days and initially involves G-proteins; the dysfunction worsens over time to finally affect all endothelial mechanisms and vascular smooth muscle. The progression of the associated intimal hyperplasia parallels the alteration in endothelial function, suggesting a permissive role of the dysfunction in the development of this acute form of coronary graft vasculopathy. PMID- 9229296 TI - Retrograde flush and cold storage for twenty-two to twenty-five hours lung preservation with and without prostaglandin E1. AB - BACKGROUND: Our previous study showed that retrograde flush through the left atrium is better than antegrade flush in 6-hour lung preservation. Whether it is feasible in long-term lung preservation is not clear. Several studies suggested that prostaglandin E1 may not be necessary in retrograde flush because of the low vascular resistance on the venous side. This study evaluates the effects of retrograde flush and prostaglandin E1 in 24-hour lung preservation. METHODS: Canine donor lungs were retrograde flushed with University of Wisconsin solution. Group A (n = 7) was pretreated with prostaglandin E1. No prostaglandin E1 was used in group B (n = 7). After flush and cold storage at 4 degrees C for 22 to 25 hours, left lung allotransplantation was performed. Measurements were taken before transplantation (baseline), and at 10, 30, 60, and 120 minutes after transplantation while the right pulmonary artery was occluded. RESULTS: After 120 minutes of reperfusion, the oxygen tension and carbon dioxide tension were 643 +/ 24 and 37 +/- 3 mm Hg in group A and 600 +/- 29 and 37 +/- 3 mm Hg in group B, respectively (p = NS). Pulmonary artery pressure (group A vs group B) was 20 +/- 1 versus 28 +/- 2 mm Hg (p < 0.01); right atrium pressure: 4 +/- 1 versus 8 +/- 1 mm Hg (p < 0.01); left pulmonary vascular resistance: 1109 +/- 51 versus 1525 +/- 133 dyne.sec.cm-5 (p < 0.05); airway resistance: 22 +/- 1 versus 24 +/- 1 cm H2O/L/sec (p = NS); lung dynamic compliance: 30 +/- 1 versus 26 +/- 1 cc/cm (p < 0.05) respectively. As compared with the baseline (19 +/- 1), airway resistance was significantly increased after 2 hours of reperfusion in group B (p < 0.05). Electron microscopy revealed that type I pneumocytes, capillary endothelial cells, and epithelial cells of bronchi were well preserved and the contents of lamellar bodies of type II pneumocyte were reduced. CONCLUSIONS: Canine lung was well preserved by retrograde flush and cold storage with University of Wisconsin solution after 24 hours preservation. Pretreatment of prostaglandin E1 is helpful in reducing pulmonary vascular resistance and airway resistance and improving lung dynamic compliance. PMID- 9229297 TI - Donor blood perfusion improves myocardial recovery after heart transplantation. AB - BACKGROUND: Improved methods of donor heart preservation may allow for prolonged storage and permit remote procurement. Previous attempts to use oxygenated perfusion circuits during storage have not gained widespread acceptance because they were either too impractical or complicated to use for remote harvest. We hypothesized that collection and perfusion of donor blood during prolonged storage may improve myocardial recovery. Our aim was to devise a safe, simple, cost-effective system that could be used in any hospital setting. METHODS: Yorkshire pigs (40 to 50 kg) were used to perform 16 orthotopic heart transplantations with either continuous perfusion with donor blood (BL, n = 8) or standard hypothermic storage (CON, n = 8). After administration of heparin, hypothermic (4 degrees C) cardioplegic arrest, and donor heart extraction, donor blood (2688 +/- 166 ml) was harvested in the BL group. After filtration for particulate matter, blood perfusion was initiated via a standard intravenous transfusion apparatus at room temperature (20 degrees C) and a pressure of 60 mm Hg and maintained during storage. Arterial and coronary sinus blood samples were obtained to examine myocardial oxygen extraction, lactate release, and acid production. A Millar micromanometer was used to measure left ventricular developed pressures at an end-diastolic pressure of 2 and 10 mm Hg both before and after transplantation. RESULTS: All pigs (eight of eight) in the BL group were successfully weaned off bypass compared to three of eight in the CON group (p < 0.01). Developed pressures (at left ventricular end-diastolic pressure = 10 mm Hg) was improved in the BL group (mean +/- SD: baseline: BL: 90 +/- 16 mm Hg vs CON: 83 +/- 12 mm Hg, p = NS; posttransplantation: BL: 66 +/- 8 mm Hg vs CON: 35 +/- 29 mm Hg, p < 0.05). Similarly, maximum dP/dt was higher in the BL group. Lactate release was higher at cross-clamp removal in the BL group (2.4 +/- 0.3 mmol/L vs 0.7 +/- 0.2 mmol/L, p < 0.01). There were no differences in oxygen extraction or acid production during reperfusion. CONCLUSIONS: Perfusion of donor blood improved the ability to wean off bypass after 4 hours of storage. Blood perfusion permitted persistent myocardial metabolism during the ischemic period, which led to improved functional recovery. Harvesting donor blood for subsequent perfusion during prolonged storage may improve the results of orthotopic heart transplantation and allow for more distant procurement of donor organs. PMID- 9229298 TI - Bronchial-pulmonary artery fistula after unilateral lung transplantation: a case report. AB - We report on a right bronchial-pulmonary artery fistula resulting in fatal hemoptysis in a 54-year-old man, 3 months after right unilateral transplantation for end-stage emphysema. The posttransplantation period was complicated by ulcerative tracheobronchial aspergillosis. Early treatment with itraconazole was performed. All samples of bronchial washing, bronchoalveolar lavage, and bronchial and transbronchial biopsy specimens were free of aspergillus after 3 weeks of treatment. Necropsy showed a fistula between the right pulmonary artery and the main right bronchus, situated just beneath the suture line. PMID- 9229300 TI - Sigmoid diverticular perforation complicating lung transplantation. AB - We present three lung transplant recipients who had sigmoid colonic diverticular perforation within 4 weeks of transplantation, giving an overall incidence of 8.6% (3 of 35) in our population. Our cases are unusual because they all occurred in the early posttransplantation period and because the incidence of perforation is substantially higher than that reported in other transplant populations. The reason for the apparent increased incidence of perforation in our lung transplant recipients is unclear, but it is likely related to the short follow-up period, intense posttransplantation immunosuppression, perioperative hypoperfusion, and increased intraluminal pressure from the use of narcotics and bowel stimulants. We discuss these potential causes and comment on preventive measures being undertaken at our program. PMID- 9229299 TI - Pulmonary artery thrombolysis and stenting after a bilateral sequential lung transplantation. AB - Bilateral sequential lung transplantation was complicated by pulmonary artery anastomotic stenosis and bilateral pulmonary thromboemboli. Pulmonary artery thrombus was eliminated by intrathrombotic but not by systemic administration of urokinase. The pulmonary emboli resulted in localized pulmonary infarctions, supporting the need for thrombolytic intervention to restore pulmonary perfusion in the absence of collateral bronchial blood flow after lung transplantation. Pulmonary artery stenosis was relieved by endovascular stenting, avoiding an early reoperative procedure. This case suggests that direct administration of thrombolytic agent may be superior to intravenous administration in the treatment of pulmonary thromboemboli. Pulmonary arterial anastomotic stenoses after lung transplantation can be relieved by endovascular procedures. PMID- 9229301 TI - Cardiopulmonary bypass during single lung transplantation. PMID- 9229302 TI - Cell-mediated responses to donor endothelium: concerning the concern. PMID- 9229303 TI - Prognosis determination in patients with advanced heart failure. PMID- 9229304 TI - Reversible causes of severe myocardial dysfunction. PMID- 9229305 TI - Pulmonary hypertension in severe congestive heart failure: how important is it? PMID- 9229306 TI - Ventricular dysfunction in the pediatric population: evaluation and intervention. PMID- 9229307 TI - Coronary artery bypass surgery in patients with left ventricular dysfunction: candidate selection and perioperative care. PMID- 9229308 TI - Mechanical circulatory assistance: changing indications and options. AB - The previously high rates of morbidity and mortality associated with implantation of LVADs for long-term support have continued to decline through design improvements, better patient selection criteria, and greater clinical experience. Patients which chronic heart failure awaiting transplantation should be considered for LVAD placement early in their clinical course, before irreversible end-organ dysfunction occurs. Both the Novacor and the vented electric HeartMate LVADs are currently being evaluated under "discharge to home" protocols for patients awaiting heart transplantation; the results will have tremendous economic and quality-of-life implications. Currently available devices have achieved a level of reliability that will allow for protocols involving device implantation as an alternative to transplantation in the near future. Physicians will then have an effective therapy available for the increasing number of heart failure patients awaiting a limited number of donor hearts, as well as for those patients not considered to be candidates for transplantation by traditional criteria. PMID- 9229309 TI - Infusional cancer chemotherapy: the critical role for nurses. PMID- 9229310 TI - Administrative and technical support of ambulatory infusional cancer chemotherapy (ICC). AB - The term "infusion" is generally equated with the parental administration of drugs in a continuous mode of delivery over some unspecified length of time. However, in terms of applicability to this paper, ICC will imply a minimum of a 24-hour delivery schedule and may include four to 7-day schedules up to protracted infusions covering many weeks. Infusional Cancer Chemotherapy programs may employ a single drug used with or without concomitant radiation therapy or admixtures of two and three drugs. Sequential alternating infusions of drug admixtures are a common mode of treatment which allow maximization of dose with minimal side effects and toxicities. The ICC model is based on the tenet that the quality of the program is directly related to the safety, efficiency, and continuity of the care provided. The supporting structures of an ICC program are rooted in four administrative and technical requisites. Strategies for program support include: 1) a team to carry out the treatment approach and to provide instructional and physical care support; 2) pre-established guidelines, protocols, policies, and procedures by which to administer the program; 3) achievement of a safe and reliable means of venous access which promotes the outpatient status; 4) a source for an accurate and reliable ambulatory infusion device and delivery system. A referral base may encompass a wide geographical area, and the patient population may be varied as to ethnicity, intellectual capacity, age, level of activity, family support and life style, thus it is important to develop a program to support the majority of patients as opposed to the carefully selected few. PMID- 9229311 TI - Ambulatory infusional cancer chemotherapy: nursing role in patient management. The Cancer Center of Boston. AB - The role of nursing in infusional cancer chemotherapy (ICC) may vary depending on the practice setting. Nurses in free-standing centers and office practices perform many duties that nurses in other facilities may not, because of the lack of many of the supports that benefit hospitals with their multidepartmental and hierarchical structures. Nurses function collaboratively with physicians in the planning and the implementation of patient treatment. Patient-related nursing responsibilities include patient/family education, drug preparation and administration, patient assessment for treatment toxicity, recognition and management of complications related to the catheter or infusion device, and telephone triage. Other duties more removed from patient care might include inventory management, research data collection and management, quality assurance and improvement, compliance with regulatory issues, and a myriad of other responsibilities. The transition of patient care to the outpatient setting has broadened the scope of nursing to include nonpatient care responsibilities due to financial constraints brought about by health care reform, changes in reimbursement patterns, and overhead required to maintain and deliver quality patient care. As a result of nursing responsibilities, it becomes paramount that the aforementioned constructs for program support are in place and that all nurses are consistently trained and have a template to follow for patient treatment and management. Nursing ability to perform patient-related tasks should be proven by formal written and practical competencies repeated annually and as procedural changes are implemented. The paragraphs to follow suggest nursing management of patients receiving ICC using a model developed at The Cancer Center of Boston (TCC). PMID- 9229312 TI - Surveillance of the patient receiving infusional cancer chemotherapy: nursing role in recognition and management of catheter-related complications. The Cancer Center of Boston. AB - Surveillance of the patient on infusional cancer chemotherapy (ICC) is paramount to patient safety. Key to this issue is diligence in monitoring for complications of infusion catheters. Nursing knowledge and awareness are essential for the early detection of such complications. The following is a composite of the manifestations of commonly experienced catheter-related complications. The incidence, detection and management issues are presented from the perspective of 15 years experience with ICC at The Cancer Center of Boston (TCC). PMID- 9229313 TI - Application of ambulatory infusion devices in infusional cancer chemotherapy: a model for nursing management. AB - Optimal support of the patient receiving infusional cancer chemotherapy (ICC) in the home setting is partially dependent on the provision of an infusional device that offers minimal complexity and maximal reliability and safety. The selection of the appropriate device is vital to positive patient outcomes and to the comfort level of both patient and care providers. This article presents issues and considerations in the selection of infusion devices and the nursing role in the management of patients utilizing such devices. Aspects of nursing management are presented from the perspective of The Cancer Center of Boston (TCC) model. PMID- 9229314 TI - Options for venous access in ambulatory care: issues in selection and management. AB - As advances in VAD technology continue, the biggest challenge becomes the ability to stay abreast of all the emerging devices and the evolutions in management and care. Various resources are available which provide guidelines and standards to enable providers in the care of patients with VADs (see Table 6). It is incumbent upon caregivers to disseminate information as it is gained through observations and experience with VADs. The literature must remain current and accessible to all caregivers in order to ensure optimal management of patients with VADs. PMID- 9229315 TI - Drug stability and compatibility in oncology care. AB - Infusional chemotherapy has been increasingly used based upon the fact that most drugs have a relatively short half-life following bolus exposure, and increasing the available drug concentration over time may maximize the antitumor effect. As a practical matter, the application of infusional chemotherapy especially in an ambulatory setting, absolutely requires that the individual antineoplastic agents be stable in solution at room temperature (or at body temperature for implanted pump systems) and that the drugs in the infusion (including antiemetics) be compatible. The capacity to mix antineoplastic agents and antiemetics (also colony-stimulating factors) in a single solution facilitates infusional combination chemotherapy. Technologic advances are on the horizon which will provide the capability of administration of multiple drugs through a single access site to allow one to utilize a single delivery source obviating the need for admixtures of drugs. However, in the interim, admixtures represent the optimal method for the delivery of multi-agent chemotherapy in the setting in which continuous infusional drug delivery for 24 hours or more is employed. The goal of continuous infusion cancer chemotherapy is to ensure delivery of an unaltered cytotoxic drug, avoiding situations that could affect the stability of the infusion admixture. With cancer chemotherapy infusion therapy being administered through oncology clinics without the benefits of a pharmacist, the nurse plays a pivotal role in the preparation of the infusion, supervision of the patient, and when applicable in providing counseling on the proper storage, handling of the cytotoxic drugs, and disposal of contaminated infusion materials. Thus, by optimizing the integrity of the chemotherapeutic infusion, the patient achieves maximum benefits of cancer chemotherapy and the level of success anticipated with oncology care. The nurse may also be involved with clinical studies involving the preparation of other combinations of infusional chemotherapy including antiemetics and colony-stimulating factors, requiring integrity of the infusion to incompatibility and instability. In order to avoid failure of the infusion through improper preparation or reconstitution of the infusion or improper storage conditions and the resulting unnecessary waste and disposal expenses, the nurse needs to be fully aware of the factors affecting the stability and compatibility of the infusion and to interact with other health professionals and the literature for the critical information related to maintaining the integrity of the cancer chemotherapy infusion. "Good intentions without good communication equals potential disaster." PMID- 9229316 TI - The French experience with infusional 5-FU in gastric and pancreatic cancer. AB - Ninety-five patients with metastatic pancreatic (n = 38) or gastric (n = 57) carcinomas were treated by 5-day continuous infusion of 5-fluorouracil (5-FU) plus cisplatin in two parallel phase II trials. 5-fluorouracil was given at a dose of 1000 mg/m2 for 5 consecutive days and cisplatin was given on day 2 at a dose of 100 mg/m2. The total number of cycles administered was 425, and each patient received a median number of four cycles. Severe toxicities were observed in around 20% of the patients and 2 toxic deaths occurred. Response rate assessed in 92 evaluable patients was 34%. The overall median survival was 8 months. There were no clinical or biological factors predictive of response, but a better survival was observed in patients with objective response and CA 19-9 blood levels lower than 100 UI/I. These two predictive factors are independent when tested in a multivariate model. The combination of infusion 5-FU plus cisplatin can be considered as a standard for the treatment of gastric metastatic carcinomas and as a promising schedule for pancreatic carcinomas. PMID- 9229317 TI - The dead end of 5-fluorouracil double modulation and promise of continuous infusion schedules in the treatment of metastatic colorectal cancer. PMID- 9229318 TI - Continuous infusion 5-fluorouracil as first-line therapy for metastatic breast cancer. AB - Previous phase II studies of continuous infusion Fluorouracil (5-FU) (CI 5-FU) in refractory metastatic breast cancer have shown modest activity with low toxicity. Its activity in a first-line setting has not been formally tested. Patients were eligible if they fulfilled the following criteria: metastatic breast cancer; measurable or evaluable disease, no prior chemotherapy in the metastatic setting; ECOG performance status of 0, 1, or 2: adequate bone marrow and liver function. Patients were treated with 5-FU 250 mg/m2 per day by continuous intravenous infusion for 5 weeks in a 6-week cycle. Treatment was continued until disease progression or unacceptable toxicity. In addition to the traditional endpoints of response, survival, and toxicity, quality of life was assessed with the Functional Living Index-Cancer (FLIC) and the Symptom Distress Scale (SDS). Twenty-one patients were enrolled. Among the 16 patients with measurable disease, the objective response rate was 44% (95% CI 20%, 68%) with CR rate 13% and PR rate 31%. The median duration of response was 37 weeks. Responses were not observed in patients with visceral (lung or liver) disease. Among all 21 patients in the study, the median time to disease progression was 12 weeks, and median overall survival was 64 weeks. Grade 1 or 2 mucosal and cutaneous toxicity were common. Only 4 patients (19%) had toxicity greater than grade 2; three patients had grade 3 mucositis, and 1 patient developed an indwelling catheter infection requiring its removal. Among responding patients, mean FLIC scores improved from 114.3 at baseline to 128.7 at week 8 (p = 0.11). Symptoms reported on the SDS generally improved in responding patients. Continuous infusion 5-FU as a first line therapy for metastatic breast cancer has moderate activity and low toxicity. Its use should be considered in the first-line setting when toxicity needs to be minimized. PMID- 9229320 TI - The EORTC GI group experience with high-dose infusional 5-FU in colorectal cancer. AB - The EORTC Gastrointestinal Tract Cancer Cooperative Group has conducted a randomized trial of high-dose infusional 5-fluorouracil (FU) with or without Methotrexate (MTX). FU was given as a 48-horus infusion of 60 mg/kg every week x 4, biweekly x 4, and subsequently every 3 weeks. Half of the patients also received 40 mg/m2 MTX as a bolus injection just prior to the FU infusion. A total of 312 patients were randomized. High-dose infusional FU was very well tolerated with virtually no haematological, renal, hepatic or cutaneous toxicity. Nausea and vomiting occurred in 35% and diarrhea in 24% of patients but was almost never severe. Cardiac toxicity and ataxia were seen in less than 5% of patients. Methotrexate lead to a significantly higher incidence of stomatitis, which was severe in 10% of patients. Eleven percent of the high-dose infusional FU patients showed an objective response with stabilization in an additional 35%; median survival was 9.3 months. With the addition of methotrexate a 23% response rate was seen (p = 0.025) and survival was 12.5 months (n.s.). We demonstrated the favorable therapeutic index tolaribility of high-dose (60 mg/kg), short-term (48 hours), frequent (weekly-biweekly) infusional FU and the ability of low-dose MTX to positivity modulate this FU treatment. PMID- 9229319 TI - Neoadjuvant chemotherapy with continuous infusion of cisplatin and 5 fluorouracil, with or without leucovorin, for locally advanced nasopharyngeal carcinoma. AB - Cisplatin-based induction chemotherapy has been extensively tested in nasopharyngeal carcinoma for the improvement of local and systemic control and survival of this disease. In this study, we report the results of the treatment with induction chemotherapy in 40 patients with locally advanced carcinoma of the nasopharynx (LANPC) with four courses of cisplatin (P) 25 mg/m2 per day and 5 fluorouracil (F) 1000 mg/m2 per day both in a 4-days continuous infusion, with or without leucovorin (L) 250 mg/m2 per day in 2-hour infusion at the beginning of daily administration of PF, followed by sequential radiotherapy. All except one were in stage IV. The overall response after induction chemotherapy was 93%, with 55% CR and 38% PR. Definitive overall response after radiotherapy was 98%, with 80% CR and 18% PR. At a maximum follow up of 11 years, the overall survival rate is 55%. Induction chemotherapy with continuous infusion of PF with or without leucovorin followed by radiotherapy is a highly active regimen for the treatment of locally advanced nasopharyngeal carcinoma with response and survival rates comparable to other combinations of sequential or simultaneous chemotherapy and radiotherapy. PMID- 9229321 TI - The Spanish experience with high-dose infusional 5-fluorouracil (5-FU) in colorectal cancer. The Spanish Cooperative Group For Gastrointestinal Tumor Therapy (TTD). AB - Background In a previous phase I to II trial, we have shown that the maximum tolerable dose (MTD) of 5-Fluorouracil (5-FU) in 48-hour continuous infusion (CI) weekly was 3.5 gr/m2. In a subsequent confirmative phase II trial with 85 evaluable patients, a 38.5% response rate was obtained and the median survival reached was 12 months. These data were comparable to those achieved by biochemical modulation of 5-FU with Leucovorin. On this basis we tried to modulate high-dose 5-FU (3 gr/m2) with oral leucovorin (LV) but the regimen was too toxic and the dose had to be reduced. A new phase II trial with 2 g/m2/week plus oral leucovorin was planned. Patients received a median dose intensity of 5 FU of 1.6 g/m2/week (range 0.9-2). Three complete responses and 36 partial responses were observed. Overall response rate was 37.5% (95% CI, 28% to 46.8%). Median time to progression has been 7.4 months, and median survival 14.4 months. WHO grade 3 diarrhea was seen in 27 patients (24.5%). Grade 3 mucositis was also seen in 9 (8.1%) patients, and grade 4 was observed in one. Grade 3 nausea and vomiting was reported in 13 (11.7%) patients. Grade 3 hand-foot syndrome was detected in only 5 (4.5%) patients. Grade 4 leukopenia was observed in 1 case and grade 3 to 4 thrombocytopenia was observed in two cases, respectively. Oral leucovorin modulation of weekly 48-hour continuous infusion of 5-FU at 2 g/m2 is an active regimen, with diarrhea and mucositis as main limiting toxicities. Its antitumor activity does not seem superior to that obtained with a weekly 48 hour continuous infusion of 5-FU alone at a dose of 3.5 g/m2. PMID- 9229323 TI - Weekly high-dose infusional 5-fluorouracil (HD-5-FU) combinations in the treatment of advanced breast cancer: results of phase I/II studies with weekly 24 hour infusion of HD-5-FU plus high-dose folinic acid (FA) alone and in combination with paclitaxel and cisplatin. AB - Our Phase II study results demonstrating high efficacy and low toxicity for a weekly schedule of high-dose 5-fluorouracil/folinic acid (HD5-FU/FA) in intensively pretreated metastatic breast cancer prompted addition of paclitaxel to this regimen in a phase I/II study in outpatients. TREATMENT: Patients were treated with HD5-FU (24-hour infusion)/FA (2h infusion prior to FU) weekly for 6 weeks (d1, 8, 15, 22, 29, 36) and Paclitaxel (3-hour infusion) was administered additionally on day 1 and day 22, q day 50. During Phase I we chose the following dose levels (dl): Fixed doses of FA d11-4 500 mg/m2 followed by HDFU, 24-hour infusion d11: 1.5, d12: 1.8, d13 and d14: 2.0 g/m2. .3-hour infusion of Paclitaxel on day 1 and day 22 d11 to d13: 135. d14: 175 mg/m2. D14 was chosen to be further evaluated during phase II. Results of an interim analysis were presented. PATIENT CHARACTERISTICS: 46 patients entered this trial during phase II and had the following characteristics: age 46 years (26 to 70) WHO performance status 0/1, metastatic disease sites 2.5(1 to 4). All patients had bidimensionally measurable disease. PRETREATMENT: 9 patients had received adjuvant chemotherapy, 16 patients prior chemotherapy for metastasis, and 21 both adjuvantly and for metastasis. Of 29 anthracycline-pretreated patients, 25 had anthracycline-resistant disease. RESULTS: We observed the following results in 35 evaluable patients: CR 3% (1/35), PR 51% (18/35), SD 40% (14/35). PD 6% (2/35). RR (Response rate) was 54%, 95% confidence interval 36 to 76%. The response concerning 20 patients with anthracycline resistant disease was: RR 55% (11/20). Median number of treatment cycles per patient was 3 (1 to 5), time to maximum response 2 months (1 to 5), remission duration 8+ months (2 to 17). Median survival time is not yet reached. CONCLUSIONS: The combination of paclitaxel with weekly HDFU/FA is well tolerated in the second-line treatment of metastatic breast cancer and indicates high efficacy also in anthracycline-resistant disease. In an ongoing phase II study, we examine the addition of cisplatin to the regimen in the first-line treatment of metastatic breast cancer. Those trials confirm that infusional weekly HD-5-FU plus folinic acid is of considerable interest in the treatment of advanced breast cancer. Randomized studies are warranted. PMID- 9229322 TI - Weekly infusional 5-fluorouracil plus/minus other drugs for the treatment of advanced gastric cancer. AB - Based on preclinical data and the promising results being achieved with infusional 5-FU in colorectal and breast cancer, we investigated a weekly schedule of a 24-hour infusion of 5-FU plus folinic acid (HD-FU/FA) in patients failing to first-line chemotherapy and HD-FU/FA plus cisplatin (C) or plus cisplatin/epidoxorubicin (C/E) in chemo-naive patients with advanced gastric cancer. In all three trials the results achieved with the tested chemotherapy regimens indicated high activity and good tolerability. All three protocols were administered as outpatient treatment. With HD-FU/FA and overall response rate of 24% and a median survival time of 5 months was observed in 17 patients refractory to or relapsing after first-line chemotherapy. HD-FU/FA/C induced an overall response rate of 66% and a median survival time of 13 months. Of note was the high activity of this regimen in patients with malignant ascites. HD-FU/FA/C/E also proved to be an interesting regimen similar active as HD-FU/FA/C but it was subjectively less well tolerated. In patients with locally advanced disease the response rate was 90% (10/11), and in patients with distant metastases 50% (8/16). PMID- 9229324 TI - The French experience with infusional 5-fluorouracil in advanced colorectal cancer. AB - The French contribution to the development of 5-fluorouracil (5-FU) infusion in advanced colorectal cancer includes five randomized trials and specific researches on the 48-hour bimonthly regimen, chronotherapy, and individual dose adaptation. The bimonthly 48-hour LV5-FU bolus and infusion is the standard treatment outside the research protocols and the control arm of the ongoing trials. PMID- 9229325 TI - High-dose infusional 5-fluorouracil combination therapy of metastatic gastric and colorectal cancer. AB - To improve the therapeutic ratio of palliative chemotherapy in patients with metastatic colorectal and gastric cancer 5-fluorouracil (5-FU) was administered as weekly high-dose 24-hour continuous infusion in combination with leucovorin (LV) and interferon-alpha-2b (IFN) as biomodulating agents: Chemotherapy consisted of a weekly schedule of 500 mg/m2 leucovorin as a 2-hour infusion, followed by a 24-hour continuous infusion of 2500 mg/m2 5-FU. IFN was administered subcutaneously at a dose of 3 mio I.E. three times a week. In patients with gastric carcinoma, etoposide (VP) 100 mg/m2 as 30-minute bolus infusion was added. Eighty-five patients (colorectal: 55 points, gastric: 30 points) are evaluable for response, toxicity, and survival analysis. Colorectal: CP+PR: 19/55 (34.5%), NC: 25/55 (45.5%), PD: 11/55 (20.0%). Median duration of remission in months (90% confidence interval): 5.2 (3.1 to 9.2), median survival since the start of salvage chemotherapy: 13.9 months (12.3 to 20.1), from initial diagnosis of metastasis: 30.2 months (26.3 to 44.5). Gastric: CR: 8/30 (26.7%), PR: 14/30 (46.6), NC: 5/30 (16.7), PD: 3/30 (10.0%). Median duration of remission in months (90% confidence interval): 6.75 (1.5 to 16.2), median survival since start of chemotherapy: 15.1 months (90% confidence interval 11.8 to 20.3 months). Hematologic toxicity: hemoglobin: I: 20.0%, II: 10.0%, leukocytes: I: 13.3%, II: 33.3%, III: 16.6%, platelets: I: 10.0% and III: 3.3%. Hematologic toxicity was moderate to negligible, peripheral toxicity consisted mainly of tolerable stomatitis and diarrhea. Dose and schedule intensified weekly 5-FU combination therapy in metastatic colorectal and gastric cancer is highly active in terms of response and median survival time. Chemotherapy pretreated patients with colorectal cancer seem to have a substantial survival benefit with this salvage protocol. PMID- 9229326 TI - Infusional 5-fluorouracil in the treatment of gastrointestinal cancers: The Royal Marsden Hospital experience. PMID- 9229327 TI - High-dose infusional 5-FU in the treatment of advanced colorectal cancer: a summary of the European experience. AB - 5-fluorouracil (5-FU) is the best available drug for the treatment of advanced colorectal cancer. A review of published data strongly suggest that with continuous infusion an enhanced treatment outcome as opposed to treatment with standard bolus injections can be obtained. This could be due to the schedule, the high dose-intensity or both. It is likely that there exists a relationship between the dose intensity of 5-FU and the response in colorectal cancer. With weekly 24 to 48 hours continuous infusion of 5-FU a very high dose intensity can be achieved. High-dose infusional 5-FU is noncross-resistent with (modulated) bolus 5-FU and the European experience with this treatment modality will be reviewed. The comparison of the activity of modulated "standard" bolus 5-FU versus high-dose infusional 5-FU, given by weekly intermittent or by protracted continuous infusion (PCI), is the subject of ongoing trials. Additional studies will answer the question whether modulation of high-dose 5-FU will result in a superior treatment outcome compared with high-dose 5-FU alone. PMID- 9229328 TI - First-line protracted venous infusion fluorouracil with CisDDP or carboplatin in advanced colorectal cancer. AB - A total of 55 patients with measurable colorectal metastatic carcinoma were studied to evaluate the impact on toxicity, response, and survival of protracted venous infusion (PVI) 5-FU 200 mg/m2 per day with Cis-DDP 80 mg/m2 or carboplatin 300 mg/m2 every 3 weeks, 1-hour infusion. Patients received continuous uninterrupted therapy until there were signs or symptoms of toxicity. Both 5-FU and cisplatin were withheld when patients experienced grade II stomatitis and diarrhea, severe nausea or vomiting not controlled by standard antiemetic therapy, and clinically significant hand-foot syndrome. The toxicity was neurological (20% grade 2 and 3) hematological (13% grade 2) and dermatological (11% grade 2). The overall response (CR+PR) was 24% with a median survival of 13 months. The results of our study show that there is no improvement in response rate, response duration or survival compared with historical trials. However, this study does confirm the valuable palliative role of the protracted 5-FU infusion treatment. Colorectal carcinoma is one of the most common neoplasms in Western societies, being second only to lung cancer as a cause of death from malignancy. The management of nonmetastatic primary disease in surgical, with adjuvant chemotherapy for those at high risk of relapse. However, for those with metastatic disease at diagnosis or recurrent disease after resection, cytotoxic chemotherapy is the treatment of choice and fluorouracil (5-FU) is the most active cytotoxic agent in this disease, with a response rate of approximately 20%. Efforts to improve the response rate have focused on the use of agents to modulate 5 FU. The Southwestern Oncology Group (SWOG) study reported by Leichman et al. (1) and a study from the United Kingdom by Hill et al. (2) compared conventional FU to modulated FU and found no improvement in response rate or survival. In the SWOG study, two different schedules of bolus FU and LV were compared with bolus FU alone and to continuous infusion FU administered alone or modulated by LV or PALA. In this study, the results obtained with bolus FU were superior to most of the studies in the literature: The response rate was 26%, and the median survival was 14 months. The high- and low-dose LV and FU groups showed response rates and survival similar to bolus FU alone. However, in 12 previously reported randomized studies comparing FU and LV or FU alone, nine reported that the combination of FU and LV produced significant increases in response rates and two reported significant increase in survival (3, 4). Many of these trials used the dose schedules reported in the SWOG trial. Protracted venous infusion (PVI) 5 FU has been shown to have superior efficacy with less toxicity in colorectal cancer when compared to bolus 5-FU and synergy between cisplatin and 5-FU has been demonstrated in vitro. Consequently, we have investigated the efficacy of the combination of bolus cis or carboplatin and PVI 5 FU in 55 patients with advanced colorectal cancer using survival, response rate, symptomatic response, and toxicity as study endpoints. PMID- 9229329 TI - CPT-11: the European clinical development. AB - CPT-11 is a camptothecin derivative with a broad spectrum of antitumor activity, both in vitro and in vivo. Like camptothecin, CPT-11 is a selective DNA topoisomerase I inhibitor. Phase I trials were conducted in Europe to determine the dose and schedule for phase II trials. These phase I trials assessed the toxicity of CPT-11 in 235 patients and tested three administration schedules: a single infusion once every 3 weeks; a weekly infusion for 3 out of 4 weeks; and a daily infusion for 3 consecutive days every 3 weeks. The maximum tolerated dose (MTD) was 115 mg/m2 in the daily schedule and 145 mg/m2 in the weekly schedule. When the drug was administered once every 3 weeks, diarrhea became the dose limiting toxicity at doses above 350 mg/m2. This schedule offered the highest dose intensity and the best tolerability profile, and was the most convenient for outpatient treatment. Finally, using this schedule, concomitant administration of high-dose loperamide allowed the dose of CPT-11 to be increased to 750 mg/m2. An ongoing phase I trial is investigating the combination of CPT-11 and 5 fluorouracil (5-FU) in various solid tumors. Although the MTD has not yet been reached, preliminary results show no pharmacokinetic interaction between the two drugs, contrary to a previous Japanese study. Based on the results of the three phase I trials, CPT-11 350 mg/m2 as an intravenous infusion over 30 minutes once every 3 weeks was recommended for phase II trials, which started in Europe in 1992. To date, CPT-11 has shown remarkable efficacy in colorectal cancer, even in patients resistant to 5-FU. Interesting results have also been obtained in pancreatic, cervical and lung cancers. Future trials will explore the place of CPT-11 in combination with other cytotoxic agents or radiotherapy. PMID- 9229330 TI - Discrepancy between in vitro and in vivo antifungal activity of albendazole. AB - Albendazole has in vitro activity against Cryptococcus neoformans and reduced in vitro activity for albendazole when compared with Candida albicans. The major metabolite of albendazole, albendazole sulphoxide showed no in vitro activity against isolates of either fungus. Immunocompetent mice infected intravenously (i.v.) with C. albicans were treated with albendazole doses of 20-600 mg kg-1 per day in noble agar or sesame oil for per oral (PO) administration, or 80 mg kg-1 per day in DMSO for intraperitoneal (i.p.) and i.v. administration for 10 days, and were observed for survival. Mice infected with C. neoformans intracranially received albendazole in daily doses of 600 mg kg-1 prepared in DMSO (i.p.) or peanut butter/rat chow (PO) for 10 days and were observed for survival. Mortality was not different between the treated and control animals in any study. Plasma samples from uninfected mice dosed with similar formulations and doses of albendazole were analysed by HPLC for albendazole and albendazole sulphoxide. No albendazole could be detected in any sample, while concentrations of albendazole sulphoxide (286-8697 ng ml-1) were observed in all samples. These data suggest that the absence of in vivo activity for albendazole is due to rapid conversion to the inactive albendazole sulphoxide metabolite. PMID- 9229331 TI - Cloning and analysis of an actin-encoding cDNA from the dimorphic pathogenic fungus Histoplasma capsulatum. AB - We have cloned an actin-encoding cDNA from the dimorphic fungus Histoplasma capsulatum, an important pathogen of humans. The predicted amino acid sequence as well as the general codon pattern of Histoplasma actin revealed the highest degree of similarity to the actin of the filamentous ascomycete Aspergillus nidulans. Southern blot analysis determined that actin was encoded by a single copy in the Histoplasma genome. Northern blot analysis showed a single 1700 nt transcript in yeast and mould cells as well as in cells undergoing the temperature induced mould-to-yeast conversion. Actin mRNA levels normalized to 18 S rRNA were found to be equivalent in all the stages examined, except for a sharp four-fold transient decrease 4 h into the mould-to-yeast conversion. These data suggest that actin mRNA would not be a suitable internal marker for expression studies during Histoplasma mould-to-yeast morphogenesis. PMID- 9229332 TI - Identification of a concanavalin A-binding antigen of the cell surface of Sporothrix schenckii. AB - Sporothrix schenckii (1099-18) cell wall peptido-rhamnomannan (CWPR) was fractionated by affinity chromatography with Concanavalin A. The Con A-bound and Con A-unbound fractions were probed with an anti-S. schenckii rabbit serum. We identified within the Con A-bound fraction three main antigens with approximate molecular weights of 84, 70 and 58 kDa. Glycopeptide beta-elimination reduced rabbit antiserum reactivity for the 84 kDa antigen (gp84) with concomittant enhanced reactivity for the 70 kDa antigen (gp70). By Western blot with Con A-HRP conjugate we demonstrated that gp84 strongly reacted with this lectin and this was the predominant antigen identified. The gp84 antigen was also demonstrated to be present on other S. schenckii strains. PMID- 9229333 TI - Molecular cloning of a Rho family, CDC42Ca gene from Candida albicans and its mRNA expression changes during morphogenesis. AB - The small GTP-binding protein family regulates various cell functions in mammalian and yeast cells. In the yeast Saccharomyces cerevisiae it has been known to be involved in vegetative growth. As an initial attempt to explore the involvement of CDC42, a member of this family, in the regulation of morphological changes in Candida albicans, we isolated a gene encoding this protein (CDC42Ca) from this fungus. The sequence of isolated gene revealed an open reading frame of 570 nucleotides with the potential to encode a protein of 190 amino acids with a predicted molecular weight of 20.5 kDa. The deduced amino acid sequence was highly homologous to CDC42s from yeast (87.8%), human (76.4%) and Caenorhabditis elegans (73.7%). The CDC42Ca mRNA level showed a transient increase with a peak at 2 h after the fresh medium shift (28 degrees C) when cells synchronously formed buds, whereas it displayed a gradual increase up to 12 h after the medium shift (37 degrees C) with elongation of germ tubes. This suggests that CDC42 may play a role in the bud emergence and also germ tube formation in C. albicans. PMID- 9229334 TI - Molecular taxonomy and GC/MS of metabolites of Scytalidium hyalinum and Nattrassia mangiferae (Hendersonula toruloidea). AB - ARDRA of ribosomal genes confirmed that the hyphomycete Scytalidium hyalinum is identical to the coelomycete Nattrassia mangiferae (Hendersonula toruloidea, with arthroconidial synanamorph Scytalidium dimidiatum). S. hyalinum may be just a melanin-less cultural mutant of S. dimidiatum. This explains why the two species are regularly found in the same patient population as agents of human dermatomycosis. Two strains from plant sources were found to be identical to the human strains in ITS-RFLP patterns. RFLP of SSU and LSU was hampered by the frequent occurrence of introns. Profiles of secondary metabolites were studied using TLC, GC and GC/MS. Production of metabolites varies considerably with the strain, but two naphthoquinone compounds were nearly always present. Selected-ion monitoring (SIM) of these compounds enabled the specific recognition of the taxon. PMID- 9229336 TI - Increased tissue resistance in the nude mouse against Candida albicans without altering strain-dependent differences in susceptibility. AB - Strain differences in tissue responses to infection with Candida albicans were examined in nude mice having susceptible (CBA/CaH) and resistant (BALB/c) parentage. Homozygous (nu/nu) mice of both strains were more resistant to systemic infection with C. albicans than heterozygous (nul+) littermates as indicated by a reduction in both the severity of tissue damage and colony counts in the brain and kidney. However, the tissue lesions in nu/nu CBA/CaH mice were markedly more severe than those in nu/nu mice with the BALB/c background. This pattern was reflected in the greater fungal burden in the CBA/CaH strain. Analysis of cDNA from infected tissues using a competitive polymerase chain reaction excluded interferon-gamma (IFN-gamma), tumour necrosis factor-alpha (TNF alpha), and interleukin 6 (IL-6) as mediators of the enhanced resistance of the nude mice. The results confirm that the different patterns of lesion severity in BALB/c and CBA/CaH mice do not involve T lymphocyte-mediated pathology, and are consistent with the hypothesis that strain-dependent tissue damage is not dependent on the effector function of macrophages or their precursors. PMID- 9229335 TI - Virulence of an aspergillopepsin-deficient mutant of Aspergillus fumigatus and evidence for another aspartic proteinase linked to the fungal cell wall. AB - A gene replacement was performed to produce mutants of Aspergillus fumigatus deficient in the aspergillopepsin PEP (E.C. 3.4.23.18). The correct replacement of the PEP gene was confirmed by PCR and Southern hybridization experiments, whereas the absence of PEP production was demonstrated by Western blots. The culture supernatant of the transformants showed no detectable acid proteinase activity, suggesting that there is only one acid proteinase secreted by the fungus. The wild-type strain and the PEP-deficient mutants invaded tissues to a similar extent and produced comparable mortality in guinea pigs. As PEP represents a third secretory proteinase of A. fumigatus and the other two proteinases also did not show significant influence on fungal invasiveness, it is probable that secreted proteinases do not contribute decisively to tissue invasion in the pathogenesis of systemic aspergillosis. However, immunofluorescence on A. fumigatus colonies using polyclonal antibodies to PEP showed a similar pattern for the wild-type and for the mutants, with a bright fluorescence on young conidiophores, on submerged mycelium and on the tips of growing aerial mycelium. Conidia and mature aerial hyphae were not fluorescent. This pattern could reflect the existence of another crossreacting aspartic proteinase (PEP2) which was found to be sensitive to pepstatin but tightly linked to the fungal cell wall. PMID- 9229337 TI - Protein synthesis patterns of Paracoccidiodes brasiliensis isolates in stage specific forms and during cellular differentiation. AB - In this paper we compared the protein synthesis patterns of Paracoccidioides brasiliensis isolates. The protein profiles were compared for both yeast and mycelial forms and similarity analysis among them was performed by calculating similarity matrices and grouping the isolates in dendrograms. The examined isolates exhibited highly variable cellular morphology at 36 degrees C, when typical yeast cells were expected. On the other hand, at 26 degrees C all the isolates showed mycelial morphology. The analysis of protein synthesis profiles made it possible to cluster the P. brasiliensis isolates into groups that correlated with the morphological data. Interestingly, growth at 36 degrees C strongly decreased the heterogeneity of protein synthesis patterns seen in mycelial isolates. It was possible to cluster the isolates grown at 36 degrees C in three groups based on their two-dimensional protein synthesis analysis. The similarity index observed among the mycelial isolates was lower than that obtained with yeast cells, suggesting a more homogenous gene expression pattern in the host-adapted form than in the saprobic phase. PMID- 9229338 TI - IgG, IgM and IgA antibody response for the diagnosis and follow-up of paracoccidioidomycosis: comparison of counterimmunoelectrophoresis and complement fixation. AB - IgG, IgM and IgA antibodies to GP43 (glycoprotein fraction of Paracoccidioides brasiliensis) were measured by ELISA in 63 samples from 23 patients with paracoccidioidomycosis before and twice after chemotherapy was started. Antibodies against P. brasiliensis were detected by indirect immunofluorescence (IF) (IgG, IgM and IgA isotypes), counterimmunoelectrophoresis (CIE) and complement fixation. Two control groups composed of 19 healthy individuals and 12 patients with other diseases (six with histoplasmosis, three with tuberculosis and three with other mycoses). The highest efficiency percentages were found with IgG and IgA-ELISA (100%), IgG-IF (96.2%), CIE (94.4%) and the lowest with CF (75.9%). Highest positive and negative predictive values (100%) were observed for IgG and IgA ELISA. IgG and IgM-ELISA antibodies are more often found in patients with acute than chronic disease (P = 0.01). Four to six months after treatment follow-up showed decreased levels of IgG and IgM-ELISA for acute cases and decreased titres of CIE for chronic cases in relation to pretreatment levels. This study suggests that IgG-ELISA anti-GP43 represents a good marker to monitor clinical response to therapy. PMID- 9229339 TI - Membrane changes associated with the early stages of apoptosis in HEp-2 cells decrease susceptibility to adherence by Candida albicans. AB - The aim of the present work was to establish whether apoptotic HEp-2 cells demonstrated an altered susceptibility to adherence by the pathogenic yeast Candida albicans. Cultures of HEp-2 cells were treated with concentrations of three chemotherapeutic agents sufficient to induce cell death by apoptosis and this was confirmed by microscopic examination and by the loss of membrane asymmetry (as indicated by phosphatidylserine externalization) and the fragmentation of nuclear DNA into distinct subunits. Cells in the early stages of apoptosis demonstrated a reduced susceptibility to adherence by a number of strains of C. albicans. A correlation between the decrease in yeast adherence and the loss of membrane asymmetry, associated with the early stages of apoptosis, was established. PMID- 9229340 TI - Unusual mould infection of the human stratum corneum. AB - Two atopic siblings presented with unilateral chronic scaly plantar lesions. Histology revealed the presence of fungi with unusual morphological aspects. Scopulariopsis brevicaulis and Phoma spp., respectively, grew on repeated cultures. Dermatophytes were absent. Such opportunistic fungal infections of the stratum corneum are extremely rare. PMID- 9229341 TI - Stereospecific formulation and characterization of sustained release ibuprofen microspheres. AB - Ibuprofen microspheres were prepared from the racemate, (+)-S and (-)-R enantiomers using waxes such as ceresine and glyceryl stearate and stereospecifically characterized. The method of preparation of the microspheres was a hydrophobic congealable disperse phase encapsulation process and variables such as particle size, wax type, enantiomeric form were evaluated. Dissolution studies were carried out by a modified USP type II method and the samples were analysed by a stereospecific HPLC assay using S-(-)-1(1) naphthylethylamine as the derivatizing agent and fenoprofen as the internal standard. The mean particle sizes of (+)-S and (-)-R enantiomers determined by microscopy/image analysis were 64 and 99 microns respectively while that of the racemate was 48 microns. Differential Scanning Calorimetry (DSC) of ibuprofen and the enantiomers showed endothermic peaks at 72 and 55 degrees C respectively. Thermograms of the physical mixture and microspheres did not show the characteristic ibuprofen peak, indicating a change in crystallinity of the drug. Powder X-ray diffraction patterns of the enantiomers and racemic ibuprofen were found to be dissimilar indicating different crystal properties. The X-ray patterns for the microspheres did not show the characteristic peaks for the drug indicating that ibuprofen may be in solid solution with the waxes. Scanning electron micrographs of the microspheres showed a generally smooth and spongy appearance for the microspheres made of compritol and glyceryl stearate. Microspheres made from the paraffin waxes had rough and hard surface characteristics consistent with the higher melting point of the waxes. Ceresine microspheres made with the enantiomers had a rougher and more porous surface compared to the microspheres made with racemic ibuprofen. Stereospecific release of the recemate from the formulations was found to be sustained (T25 of 4 h), while release from the enantiomers was less sustained (T50 of 2 h). From the S:R ratios and statistical analysis of the data, the release of the enantiomers of ibuprofen from the formulations containing the racemate was found to be non-stereoselective. PMID- 9229342 TI - 99mTc-labelled diphytanoylphosphatidylcholine liposomes: in vitro and in vivo studies. AB - The uniquely structured diphytanoylphosphatidylcholine (DphPC) forms liposomes more stable than conventional straight chain phospholipids. In this study DphPC and pegylated DphPC (DphPC-PEG) liposomes were radiolabelled and evaluated in vitro and in vivo. 99mTc-DphPC liposomes were found to be nontoxic to human white blood cells in vitro. In addition 99mTc labelled DphPC-PEG liposomes were evaluated as a nonspecific infection imaging agent in a mouse model. Infection sites were imaged within 30 min postinjection, and the radiopharmaceutical exhibited a remarkable in vivo stability. As their biodistribution and pharmacokinetic patterns can be size-modulated, DphPC-based lipsomes are excellent candidates for diagnostic and therapeutic applications. PMID- 9229343 TI - Modelling of gas transport function of erythrocytes in haemoglobin sorption immobilization. AB - Colloid-disperse systems based on haemoglobin sorption immobilization in reticular polyelectrolytes are proposed and investigated. The efficiency of oxygen transport of these systems is much higher than that of native haemoglobin and is comparable with the efficacy of gas transport of erythrocytes. This is believed to be due to highly selective sorption immobilization of haemoglobin in microdisperse forms of permeable carboxylic reticular polyelectrolytes. Microparticles exhibit high local concentration of haemoglobin, the protein mass being by one order of magnitude higher than that of the polymer carrier. PMID- 9229344 TI - Formulation and preparation of controlled release pellets of salbutamol by the air suspension technique. AB - A controlled release preparation of salbutamol may improve patient compliance, minimise side effects and be valuable in the treatment of nocturnal asthma by extending drug action throughout the night. The aim of the study was therefore to formulate a controlled release pellet preparation of salbutamol via the air suspension technique. The study established that curing for 24 h at 38 +/- 0.5 degrees C was necessary for homogeneous film coats of Eudragit RS30D and hence stable drug release characteristics. Pellets coated with 6% Eudragit RS30D (polymer), 12.5% triethyl citrate (plasticiser) and 0.5% magnesium stearate (antitackiness agent) displayed desirable controlled drug release characteristics over the 8 h testing period. The manufacturing conditions employed in the study were shown to be reproducible thus ensuring reproducibility of drug release characteristics between batches of salbutamol controlled release pellets. Short term stability testing on the newly formulated pellets indicated no significant change in drug release characteristics relative to the initial drug release data when stored for 8 weeks at room temperature 20 +/- 2 degrees C or 37 degrees C with 80% Relative Humidity or at low temperature (5 +/- 1 degrees C). However, pellets stored at 40 degrees C showed a slower in-vitro drug release after 8 weeks of storage and therefore failed to maintain their initial drug release profile. PMID- 9229345 TI - Effect of cyclosporine A formulations on bovine corneal absorption: ex-vivo study. AB - The purpose of this study was to evaluate, in an ex-vivo study, the absorption of cyclosporine A on bovine cornea after 24 h contact with various drug delivery systems containing 1% cyclosporine A and in comparison with an olive oil formulation as the reference vehicle for cyclosporine A. The different formulations studied were poly(acrylic acid) polymeric gels in aqueous/non aqueous solvents, polyisobutylcyanoacrylate nanocapsules, and a combination of both formulations. The histological effects of these formulation on corneal cells after 24 h of contact were also studied. The lowest absorption rate of cyclosporine A was found using olive oil with a percent absorption of 2.52 +/- 1.52% (259 +/- 171 micrograms/g cornea). The three formulations developed for this study, nanocapsules, poly(acrylic acid) polymeric gel and nanocapsules gel showed significantly better absorption of CsA than olive oil, with a mean percent absorption of 5.81 +/- 2.04% (621 +/- 218 micrograms/g cornea), 6.09 +/- 2.93% (651 +/- 313 micrograms cornea) and 7.92 +/- 2.55% (847 +/- 273 micrograms/g cornea) respectively. As we studied the penetration of cyclosporine A into the different layers of the cornea, we observed that for all formulations, CsA remained at the corneal surface and did not penetrate the whole cornea. The histological study showed that olive oil, nanocapsules and poly(acrylic acid) gel in aqueous/non-aqueous solvents show some modifications on the cornea, contrary to the nonocapsules gel which did not indicate any toxic effect. The nanocapsule gel, with the highest percent absorption along with its margin of safety on the cornea, seems to present a new promising drug delivery system for ocular administration. PMID- 9229347 TI - Dielectric and thermodynamic properties of biodegradable poly(D,L-lactide-co glycolide) and the effect on the micro-encapsulation and release of captopril. AB - The mechanical and dielectric properties of three kinds of poly(lactic acid coglycolic acid) (PLG) with different molecular weights and polydispersities fractioned by ultrafiltration were investigated by dynamic mechanical thermal analysis (DMTA) and dielectric measurement. All samples showed typical behaviour of amorphous polymer under different fields. Two relaxation processes were found, a secondary relaxation in glassy state at low temperature and a glass transition relaxation. The molecular weights and polydispersities of PLGs influenced significantly both relaxation, especially the relaxation strength and location. The strength of secondary relaxation was reduced and the glass transition shifted to a higher temperature when the molecular weight of PLG increased and the polydispersity decreased. The shift of glass transition temperature (Tg) might decrease the motion of the macromolecules and resulted in a higher moduli of rubbery PLG at the temperature of the drug system (37 degrees C) and lowered the diffusivity of the drug in polymeric matrix and then the initial burst and fast diffusional release of captopril from commercial PLG were improved. PMID- 9229348 TI - Variation of droplet sizes during the formation of microcapsules from emulsions. AB - The size distributions of microcapsules are important in determining the surface area over which the contents of the microcapsules are released. The size distribution is generally not normal around the mean, but biased toward smaller capsule sizes, forming a mode around some characteristic diameter. In order to assess the factors effective in determining this characteristic drop size, size distribution analyses were carried out photographically at each stage of the complex coacervation process. In addition, the emulsion drop size distributions were investigated as a function of stirring rate, stirring time and phase ratio. The results agreed qualitatively with the theory of isotropy but were an order of magnitude smaller. This theory is valid for Newtonian liquids. Elongational stresses could be operative with large-sized macromolecules and a mechanism was proposed based on variation of elongational stresses with velocity and rates of extension. Elongational viscosities measured for the gelatin solutions supported the proposed mechanism. PMID- 9229346 TI - Retention of trypsin activity in spermine alginate microcapsules. AB - We have previously shown virus particles encapsulated in aqueous spermine alginate constructs retain immunogenicity and infectivity both in vitro and in vivo. However, because virions are complex structures with multiple reinforcing components, it was uncertain if isolated single proteins would retain functional integrity when similarly encapsulated. To examine this question trypsin, used as a model protein, was blended with aqueous sodium alginate and the blend was dispersed as fine droplets in aqueous spermine hydrochloride to generate self assembling, trypsin-containing microcapsules. Trypsin was assayed spectrophotometrically for retention of enzymatic activity using N-alpha-p-tosyl L-arginine methyl ester as substrate. Neither of the encapsulating reagents alone inhibited enzyme activity. Enzyme that escaped capture was assayed directly in the manufacturing supernatant. In mass balance studies we found that about 20-30% of activity was retained in intact capsules with the remainder resident in the aqueous manufacturing supernatant and washes. However, we found that the capsule wall appeared to inhibit enzyme activity by retarding substrate diffusion into and product diffusion out from the capsules, as evidenced by an increase in activity on lysis. Thus, it is clear that a single protein, as represented by trypsin, can retain functional integrity when encapsulated in this all aqueous system. PMID- 9229349 TI - Dynamic swelling behaviour of gelatin/poly(acrylic acid) bioadhesive microspheres loaded with oxprenolol. AB - The dynamic swelling of gelatin/poly(acrylic acid) microspheres loaded with oxprenolol has been evaluated. The movement of two distinct and characteristic swelling boundaries was measured directly using an optical microscope. Swelling rate constants associated with the inner moving front and the outer swelling boundary were determined. A polynomial equation incorporating both Fickian diffusion and case II relaxational models was used to fit the kinetics of liquid uptake. The influence of the concentration of the cross-linking agent, the poly(acrylic acid) load, the pH of the swelling medium and the particle size of the microspheres, on the swelling kinetics, was evaluated and discussed. PMID- 9229350 TI - Oxprenolol release from bioadhesive gelatin/poly(acrylic acid) microspheres. AB - Drug release from gelatin/poly(acrylic acid) oxprenolol-loaded microspheres has been evaluated using an in situ sink immersion method and a wetting method. The kinetics of drug release were analysed by applying the empirical exponential equation and by the calculation of the approximate contribution of the diffusional and relaxational mechanism to the anomalous release process by fitting the data to the coupled Fickian/Case II equation. The influence of glutaraldehyde cross-linking agent concentration, the poly(acrylic acid) content, the pH of the release medium and the particle size of the beads on the drug release kinetics were evaluated and discussed. PMID- 9229351 TI - Endocrine and behavioural responses to noise stress: comparison of virgin and lactating female rats during non-disrupted maternal activity. AB - The behavioural and endocrine responses to a 10 min white noise stress have been characterized in female virgin and undisturbed lactating Sprague-Dawley rats. Animals were continuously video-taped and frequent blood samples were collected using an automated sampling system. Noise stress caused hypothalamo-pituitary adrenal (HPA) activation, as indicated by a rapid increase in plasma corticosterone and ACTH in the virgins: corticosterone concentrations peaked 20 min after initiation of the stress before declining rapidly back to basal concentrations. In contrast, noise stress had no significant effect on either plasma corticosterone or ACTH concentrations in the lactating animals. However, 72 h after weaning the corticosterone response of the ex-lactating rats was of comparable magnitude, but longer duration to that seen in the virgins. Plasma prolactin concentrations were significantly higher in the lactating animals and declined in response to the noise whereas, a transient but reproducible increase was seen in the virgin group. In situ hybridization revealed a significantly lower basal expression of CRF mRNA in the paraventricular nucleus of lactating rats as compared to the virgins, but noise stress had no further effect. Virgin animals showed behavioural responses to the stress, including an increase in the total activity, exploratory behaviours (rearing) and displacement behaviours (grooming). Lactating animals also showed behavioural responses to the noise, but their activities were principally directed towards the pups. These data show that although lactating rats showed normal behavioural reactivity to a psychological stress they showed no statistically significant activation of the HPA axis, suggesting a dissociation of behavioural and neuroendocrine responses to this mild stress. PMID- 9229352 TI - The role of aromatization in the restoration of male rat reproductive behavior. AB - This study assessed the role of aromatization in the expression of male reproductive behavior by testing the effects of the aromatase inhibitor, fadrozole, on the restoration of male sexual behavior and partner preference in testosterone-treated gonadectomized rats. We measured nuclear estrogen receptor occupation to determine whether fadrozole blocked brain aromatase. In addition, nuclear androgen receptor assays were used to verify that fadrozole does not block androgen receptors. Mini-osmotic pumps fitted to brain infusion cannulas were used to deliver fadrozole (20 micrograms/day) into the right lateral ventricle. The majority of animals receiving fadrozole treatment with two, 10 mm testosterone filled Silastic capsules (T/F group) failed to display any sexual behavior 7 and 13 days following implant surgery. In contrast, animals receiving fadrozole treatment which were implanted with two, 10 mm testosterone capsules and one, 5 mm 1% estradiol capsule (T/F/E group) copulated normally, indicating that fadrozole's inhibition of male sex behavior was specifically due to blocking aromatase activity. Moreover, the animals which received only one, 5 mm 1% estradiol capsule (E group) also failed to exhibit male sexual behavior. Partner preference for either a sexually receptive female or a non-receptive female was measured in a three chambered apparatus for an index of sexual motivation. Repeated measures contrasts on the group x test interaction indicated that the T/F group was not significantly different from the T group. In addition, the E group did not show a preference for the receptive females and was significantly different from the T group. Fadrozole treatment resulted in a 59% decrease in brain nuclear estrogen receptor occupation relative to the T group. Fadrozole had no significant effect on brain nuclear androgen receptor occupation. Our results lend support to the hypothesis that both androgen receptor activation and aromatization are necessary for the restoration of male sexual behavior in rats. However, we found that estradiol is neither necessary nor sufficient for the restoration of partner preference. PMID- 9229353 TI - Characterization of GH3 cells overexpressing basic fibroblast growth factor (FGF 2). AB - Basic fibroblast growth factor (FGF-2) is not only a potent mitogen for various cells but also a multifunctional factor with angiogenic and chemotactic activity, and the capacity to induce the synthesis of various proteinases and to modulate endocrine function. To clarify the role played by FGF-2 in the progression of pituitary tumor, we fused rat FGF-2 cDNA to the promoter SR alpha, consisting of the early promoter of SV40 and HTLV(I)-LTR, and we cotransfected GH3 cells with pSV2-neo by an electroporation method. After selection by G418, we obtained 7 neomycin-resistant clones. Southern blot analysis of genomic DNA revealed the presence of transfected rat FGF-2 cDNA in 4 of the 7 clones. To measure FGF-2 molecules, we established a new immuno-fluorometric assay system, using 3 monoclonal antibodies against different portions of human FGF-2. This assay had a minimum sensitivity of 10 pg/ml and cross-reacted neither with acidic fibroblast growth factor (FGF-1) nor insulin-like growth factor 1 (IGF-1), even at a concentration of 100 ng/ml. Although FGF-2 was undetectable in the culture medium of any of the clones, the cell homogenate contained a significant amount of FGF-2 (7.2 ng/mg protein) in 1 of the 4 FGF-2-transfected clones (GH3FGF(+)), whereas FGF-2 was not detected (< 5.2 pg/mg protein) in the cell homogenates of either the parent GH3 cells or the control cells transfected with pSV2-neo alone (GH3FGF(-)), GH3FGF(+) grew as adherent cells and formed epithelial sheets with a growth rate similar to that of control cells. The amount of prolactin(PRL) released by TRH was greater in GH3FGF(+) than that in GH3 or GH3FGF(-). On the other hand, the sensitivity to SRIF was increased in GH3FGF(+) compared with that in other clones. The findings of these in vitro studies indicate that FGF-2, if it is expressed in pituitary tumor cells, plays little if any role in cell growth but may modulate certain cell functions such as responsiveness to hormones. PMID- 9229354 TI - Distribution of Fos immunoreactivity in the lamina terminalis and hypothalamus induced by centrally administered relaxin in conscious rats. AB - The effect of intracerebroventricular (ICV) injections of synthetic human or rat relaxin (25 or 250 ng) on the distribution of Fos detected immunohistochemically in the rat forebrain was investigated. Following ICV relaxin, many Fos-positive neurons were observed in the periphery of the subfornical organ, dorsal part of the organum vasculosum of the lamina terminalis, throughout the median preoptic nucleus, supraoptic nucleus and hypothalamic paraventricular nucleus. Such effects did not occur following ICV injection of artificial cerebrospinal fluid or the separated A and B chains of relaxin, nor following the intravenous injection of 250 ng of relaxin. Both vasopressin and oxytocin containing neurons identified immunohistochemically in the supraoptic and paraventricular nuclei exhibited Fos following ICV relaxin, and many neurons in the medial parvocellular part of the paraventricular nucleus contained Fos. The results indicate that centrally administered relaxin may increase neuronal activity in regions of the hypothalamus and lamina terminalis which are associated with cardiovascular and body fluid regulation and oxytocin secretion. PMID- 9229355 TI - Ontogeny of growth hormone and LH beta-, FSH beta- and alpha-subunit mRNA levels in the porcine fetal and neonatal anterior pituitary. AB - The mechanism underlying the ontogenetic increase in plasma growth hormone (GH), luteinizing hormone (LH) and follicle stimulating hormone (FSH) concentration during fetal life in mammalian species and the prenatal sex difference in these hormones in some species is not fully understood. To this end anterior pituitaries were collected from German Landrace fetuses and piglets at day (d) 50, 65, 80, 95, 110 pc and d 6 pp and pituitary GH, LH beta, FSH beta and alpha subunit mRNA levels were determined by measuring Northern blot hybridization signals. GH mRNA was detected in both sexes as early as d 50 pc. The mRNA level markedly increased with age in both sexes (males > females, P < or = 0.05) reaching its maximum at d 95/110 pc. LH beta mRNA signals were first detected at d 50 pc in females and at d 65 pc in males increasing thereafter to a maximum at d 6 pp in both sexes (P < or = 0.05). In males the augmentation in LH beta mRNA was delayed compared to females (P < or = 0.01). Before d 80 pc no FSH beta mRNA hybridization signals were apparent. Thereafter the mRNA level continuously increased with age (P < or = 0.01) in both sexes reaching its maximum at d 6 pp. The FSH beta mRNA level in females was always higher than in males (P < or = 0.01). As early as d 50 pc the alpha-subunit mRNA level was high in both sexes and further increased without sex difference to d 6 pp (P < or = 0.05). In conclusion, the mRNA levels of GH, LH beta and FSH beta are age and sex dependent during fetal development. We suggest that the fetal increase in plasma GH concentration can be accounted for by changes in GH mRNA levels, while the dramatic perinatal decrease in plasma GH concentration seems to be primarily controlled at the posttranslational and/or secretion level. The fetal sex difference and the increase in plasma LH and FSH concentrations seems to be primarily dependent on the cellular concentration of the gonadotropin beta subunit mRNAs and/or number of gonadotrophs. PMID- 9229356 TI - Effects of photoperiod duration and melatonin signal characteristics on the reproductive system of male Syrian hamsters. AB - Three experiments tested effects of photoperiod and the pineal hormone melatonin (MEL) on reproductive function among male Syrian-hamsters. In Experiment 1, hamsters were exposed for 32 weeks to 1 of 4 short photoperiods which varied in duration (11.5 L; 10 L; 8 L; 6 L). A fifth group was shifted from 11.5 L to 6 L after 6 weeks. Shorter photoperiods were associated with more rapid regression of the testes, but all groups eventually regressed to the same extent. In contrast, the temporal profile of testicular recrudescence, expressed as males became photorefractory, was not significantly different between groups. A decrease in photoperiod from 11.5 L to 6 L after 6 weeks did not delay the onset of recrudescence. The 11.5 L group was subdivided at week 32 and transferred to either 13 L or 16 L for the next 8 weeks to break photorefractoriness. Upon subsequent exposure to 8 L, both subgroups regressed their testes in similar fashion over weeks 40-52, indicating that the two long photoperiods were equally effective in breaking photorefractoriness. Nevertheless, FSH and prolactin were more consistently suppressed in the 16 L group following the switch to 8 L. Experiment 2 tested whether differing durations of MEL, administered s.c. each night for 9 weeks, elicit graded rates of reproductive regression in pinealectomized males. Testicular regression was more rapid in the group receiving MEL for 12 h than it was in the group receiving MEL for 8.5 h, thus supporting the hypothesis that the faster rates of testicular regression in the shorter photoperiods of Experiment 1 were due to their concomitant longer durations of nightly MEL secretion. Experiment 3 tested the hypothesis that rates of testicular regression in males receiving exogenous MEL would be affected by their prior photoperiodic history. Males were exposed to 18 L or 14 L for 7 weeks, then pinealectomized and administered 9.5 h MEL infusions s.c. each night for 9 weeks. In contrast to predictions, photoperiodic history had only transitory effects on MEL-induced testicular regression. Although the differences in MEL duration that accompany different short photoperiods have reproductive consequences (Experiment 1), the extent to which MEL duration expands during the transition from stimulatory to inhibitory photoperiods appears to be a less significant variable (Experiment 3). PMID- 9229357 TI - Differential LHRH secretion, dye coupling, and protein expression in two morphologically distinct cell types identified in GT1-7 cultures. AB - The immortalized neuronal cell line, GT1-7, has been shown to secrete LHRH in a pulsatile manner and to possess many other characteristics of hypothalamic LHRH neurons in vivo, and thus provides a potential model system for studying biochemical and physiological mechanisms regulating LHRH secretion. In the present study, two morphologically and functionally distinct types of cells have been identified in GT1-7 cultures and each type purified to over 95% homogeneity. One type (N cells) appeared more neuronal with extended neurites and somewhat rounded cell perikarya, while the other type (G cells) had flatter cell perikarya that contained filopodia but no neurites. Growth properties of the two cell types also differed. The doubling time for proliferation of N cells was nearly two-fold shorter than that for G cells and N cells displayed 'piling up' whereas G cells exhibited contact inhibition. Functionally, N cells, but not G cells, were dye coupled as measured by a fluorescence photobleaching assay. While both cell types expressed LHRH, N cells released significantly higher levels of LHRH into the culture media and exhibited more intense LHRH immunostaining. The two cell types also showed differences in immunostaining for other proteins. N cells, unlike G cells, immunostained positive for neuron-specific enolase (NSE), whereas G cells, unlike N cells, stained immunopositive for vimentin. Both cell types expressed SV 40 T antigen protein, indicating that they were derived from the same transgenic mouse hypothalamic tumour. The physiological significance of these two cell types in GT1-7 cultures remains to be determined, but elucidation of their morphological and biochemical properties is intended to contribute to better understanding and application of this experimentally important neuroendocrine cell line. PMID- 9229358 TI - Slow wave sleep drives inhibition of pituitary-adrenal secretion in humans. AB - During the first half of nocturnal sleep, the secretory response of the pituitary adrenal axis to either CRH or vasopressin (VP) administration is reduced. Two experiments were performed aiming (i) to investigate the impact of sleep on the response to a combined CRH/VP administration and (ii) to specify the onset of sleep associated pituitary-adrenal suppression and its relation to specific sleep stages. In experiment I, we compared the effect of simultaneous administration of VP (0.5 IU i.v., within 6 min) and CRH (50 micrograms bolus i.v., in the third min of VP infusion) on the secretion of ACTH, cortisol and GH in healthy men during the first nocturnal epoch of slow wave sleep (SWS) and during nocturnal wakefulness. The increase of ACTH and cortisol concentrations after combined VP/CRH administration was distinctly higher during wakefulness than sleep (P < 0.01). In experiment II, CRH (30 micrograms/h, after an initial bolus of 30 micrograms) was continuously infused in 7 healthy men on 2 nights. On one of the nights, the men were allowed to sleep (between 23.00 h and 05.00 h) after a 3-h period of wakefulness, on the other night they stayed awake throughout the experiment. In both conditions, CRH enhanced ACTH/cortisol plasma levels. Compared with concentrations during continuous wakefulness, sleep and in particular SWS was associated with a suppression of ACTH/cortisol levels (P < 0.05). The findings further support an inhibitory influence of early nocturnal sleep on pituitary-adrenal activity. The effect appears to be strongest during SWS and is probably mediated via hypothalamic secretion of a release inhibiting factor of ACTH. PMID- 9229359 TI - Lupus and cerebrovascular accidents. PMID- 9229361 TI - The effect of OM-89 (Subreum) on the murine model of systemic lupus erythematosus MRL-lpr/lpr. AB - OM-89 (Subreum), an E. coli extract, is used clinically in the treatment of rheumatoid arthritis. In this study, the authors examined the effect of OM-89 on some aspects of SLE in the murine model MRL-lpr/lpr. Animals were given OM-89 orally at a dose of 400 mg/kg weight (40 mg active substance) 5 d a week from six weeks old. It was found that mice receiving the drug reached the point of 55% (6/11) survival at the age of 33 weeks compared with 27 weeks for the control group (54%; 7/13). There was a significant increase in the delay before developing alopecia in the treated group (P < 0.01). The increase in proteinuria in the control group was significantly higher than in the treated group (P < 0.03). In a second set of experiments sacrificing the animals at week 22, a significant decrease in anti-dsDNA auto-antibodies was also found in the treated group (P < 0.05), histopathologically a less severe tubular destruction in the kidney was observed in the treated group. It can be concluded that the oral treatment of OM-89 can significantly reduce the severity of SLE in this strain of mice. It can be postulated that the administration of the bacterial extract could modulate the immune response, modifying the Th1 and Th2 balance and inducing oral tolerance. PMID- 9229362 TI - Autonomic nerve dysfunction in systemic lupus erythematosus: evidence for a mild involvement. AB - Neurologic manifestations are known to occur in patients with systemic lupus erythematosus (SLE) and significantly affect the clinical course of the disease. Nevertheless, the prevalence, pattern and severity of autonomic impairment in such patients have yet to be defined. In the present study a series of 38 female SLE patients was assessed for the presence of autonomic dysfunction. Five noninvasive standardized cardiovascular reflex tests were used. The grading system proposed by Ewing and Clarke was applied to classifying autonomic impairment according to severity. Seventeen out of 38 patients, that is 44.7%, had evidence of autonomic impairment. Most of the patients had a mild degree of dysfunction. No correlation was found for the duration of the disease while an apparent lack of the commonly described chronological sequence of autonomic involvement was observed. We suggest that in SLE patients the prevalence of autonomic impairment, when investigated, does not significantly differ from that of other SLE-associated neurological events. The contribution of a direct immunological damage to components of neural pathways in the pathogenesis of the autonomic involvement can be postulated. Clinical consequences of autonomic impairment in patients with systemic lupus erythematosus need to be elucidated. PMID- 9229360 TI - Prevalence of 13 autoantibodies and of the 16/6 and related pathogenic idiotypes in 465 patients with systemic lupus erythematosus and their relationship with disease activity. AB - With a cross sectional study of 465 consecutive systemic lupus erythematosus (SLE) patients tested for 13 autoantibodies (Aab) and two idiotypes we determined the prevalence of Aab according to disease activity, both general and at particular organ systems. Seventy seven percent of SLE sera had at least one Aab and 56% had it at high titres. Pathogenic idiotypes had a prevalence of less than 10% and 166 sera had Aab to 5 or more antigens and 9 sera had Aab against all 13 antigens tested. Patients with active disease had increased prevalence of Aab to DNP, ssDNA, ENA, mitochondria and histones when considered at 5 s.d. above the mean of normal controls. The higher positivity of Aab in patients with active disease was confirmed in logistic regression analysis adjusted by age, disease duration, and intensity of treatment. A trend was observed of increased prevalence and titres of Aab from inactive disease without treatment, to inactive disease but still being treated, to active disease. Only 22% of patients with active disease had no Aab and the higher the number of Aab the higher the frequency of active disease. Patients with active arthritis, and to a lesser degree those with active mucocutaneous involvement, had higher prevalence and titres of most autoantibodies than patients with disease activity at other organ systems. Active renal disease associated only with anti-dsDNA, whereas active CNS disease associated with anti-mitochondrial Aab. Our findings support the vision of SLE as an immune dysregulation leading to polyclonal B cell activation with resulting production of multiple Aab. Their profiles seem influenced by genetical, hormonal and environmental factors and, in turn, they contribute to the clinical picture in each patient. Disease activity influences the presence of some, but not all, Aab and some of them may remain present in some patients, even in remission. PMID- 9229364 TI - CD69 to CD3 ratio of peripheral blood mononuclear cells as a marker to monitor systemic lupus erythematosus disease activity. AB - CD69 is an early T cell activation marker. In order to investigate whether the disease activity of systemic lupus erythematosus (SLE) is correlated with T cell hyperactivity as well as B cell hyperactivity, we measured the CD69 to CD3 ratio (CD69/CD3) of peripheral blood mononuclear cells (PBMCs) from 42 SLE patients and 18 healthy controls. To assess B cell activation, we measured the levels of anti dsDNA, complement C3 and C4. Disease activity was assessed with the SLE disease activity index (SLEDAI) score. The mean value (+/- standard deviation) of CD69/CD3 for SLE patients (3.34 +/- 0.486) and healthy controls (0.92 +/- 0.015) were significantly different (P < 0.05). CD69/CD3, anti-dsDNA, C3 and C4 were all significantly correlated with SLEDAI (r = 0.50, 0.51, -0.68, -0.31, respectively, P < 0.05). CD69/CD3 could explain 25.4% of the variance in SLEDAI, and 6.7% of the variance that was independent of anti-dsDNA, C3 and C4. In patients with SLEDAI scores which were discordant with anti-dsDNA, C3 and C4 values, CD69/ CD3 still showed high correlation with SLEDAI. We conclude that CD69/CD3, which detects T cell activation is correlated with SLEDAI scores. Thus, T cell activation contributes part of the disease activity independent of B cell activity. This marker can complement in the estimation of disease activity when levels of anti-dsDNA, C3 and C4 fail to show good correlation. PMID- 9229363 TI - Pulmonary hemorrhage in systemic lupus erythematosus. AB - In order to assess the clinical characteristics and survival rate of pulmonary hemorrhage (PH) in systemic lupus erythematosus (SLE), we studied SLE patients who developed this complication. We found 34 patients within a total lupus cohort of 630 patients. All the patients had severe respiratory failure. We identified three different treatment regimens: (a) oral prednisone (1 mg/kg); (b) conventional methylprednisolone (3 g total dose) and (c) massive methylprednisolone (> 4 g). The overall survival rate was 38.2% and it was correlated with the massive regimen and early treatment (within the first 48 h after the onset of the acute event). PMID- 9229365 TI - Association between IgM anticardiolipin antibodies and deep venous thrombosis in patients without systemic lupus erythematosus. AB - Patients with systemic lupus erythematosus (SLE) are at risk of developing deep venous thrombosis (DVT). Should anticardiolipin antibodies (aCL) be detectable, this risk is significantly raised, particularly when these autoanti-bodies are cofactor-dependent. We conducted a cross-sectional study of consecutive unselected outpatients referred for clinical suspicion of DVT, as an attempt to address the following questions: firstly, were aCL antibodies associated with DVT in non-SLE patients? Secondly, was this association related to the cofactor dependence? From March 1992 to February 1994, 208 patients were enrolled in the study. Venography was positive in 110 patients (DVT patients), while the diagnosis of DVT could not be confirmed in the remaining 98 (referred to as disease controls). ACL was measured by ELISA, for IgG and IgM isotypes in two ways: fetal calf serum or bovine serum albumin were used as blocking agents to distinguish between cofactor-dependent and cofactor-independent antibodies. Positive aCL was defined as optical density (OD) values greater than the 95th percentile of OD distribution of 60 healthy controls. We found a high frequency of positive IgG aCL antibodies in both DVT patients and in disease controls (25.5 vs 23.5%). We suggest an association between IgM aCL and DVT. This association was, however, not dependent on the cofactor requirement. PMID- 9229366 TI - C2 deficiency in blood donors and lupus patients: prevalence, clinical characteristics and HLA-associations in the Brazilian population. AB - The objective of the present study was to investigate the prevalence, clinical characteristics, and HLA association of C2 deficiency in the Brazilian population. The frequency of C2 deficiency profile (C2Q degree profile) was 2.2% among 1503 blood donors and 6.6% among 166 patients with systemic lupus erythematosus (SLE). A higher incidence of clinical manifestations possibly related to immune complex disease was observed among blood donors with C2Q degree profile and their relatives with C2Q degree profile when compared to the normal C2 relatives. The comparison of clinical and laboratory features between SLE patients with C2Q degree profile and those with normal C2 revealed earlier disease onset, higher frequency of oral ulcerations and lower frequency of anti native DNA antibodies in the first group. The HLA study conducted on 18 individuals with C2Q degree profile (11 blood donors and 7 SLE patients) confirmed the previously reported association with the antigens HLA-A25, B18 and DR2, supporting the concept that probably most C2 deficiency cases, throughout the world, are due to a single mutation in the C2 gene in linkage disequilibrium with the A25B18DR2 haplotype. PMID- 9229367 TI - Risk for venous thrombosis related to antiphospholipid antibodies in systemic lupus erythematosus--a meta-analysis. AB - OBJECTIVE: To describe the relative risk for venous thrombosis (VT) associated with antiphospholipid antibodies (aPL) in systemic lupus erythematosus (SLE). DESIGN: Systematic review and meta-analysis of 26 articles that examined the association between aPL and VT in SLE. SETTING: Mostly secondary and tertiary referral centres. PATIENTS: 2249 patients with SLE, 1120 tested for LA (lupus anticoagulant) and 1563 tested for aCL (anticardiolipin antibodies). MAIN OUTCOME MEASURES: A summary of study characteristics and a critical appraisal of study quality were done. Two statistical combinations of 18 primary studies that examined the association of VT and LA and of 14 studies that examined the association of VT and aCL were performed to estimate the risk for VT associated with aPL. RESULTS: The odds ratios of the risk of VT related to the LA summarized from 18 studies were 5.61 [95% CI; 3.80-8.27] overall, 6.32 [CI; 3.71-10.78] for deep venous thrombosis and pulmonary embolism, 11.6 [3.65-36.91] for recurrent venous thrombosis after the first event. The odds ratios of the risk of VT related to aCL summarized from 14 studies were 2.17 [95% CI; 1.51-3.11] overall, 2.50 [CI; 1.51-4.14] for deep venous thrombosis and pulmonary embolism, 3.91 [1.14-13.38] for recurrent venous thrombosis after the first event. CONCLUSIONS: Patients with SLE and LA are at approximately six times greater risk for VT than patients without LA, whereas patients with SLE and aCL are approximately two times greater risk for VT than patients without aCL. We have identified important methodologic limitations and differences in study characteristics. Other risk factors for VT have not been thoroughly evaluated in these studies. Further studies are needed that provide an accurate estimate of the absolute risk for aPL related VT. PMID- 9229368 TI - Primary antiphospholipid syndrome emerging following thymectomy for myasthenia gravis: additional evidence for the kaleidoscope of autoimmunity. AB - The etiology of autoimmune diseases is multifactorial. In many of them the stimulation by a specific autoantigen is claimed to be responsible for the initiation of the disease. Alternatively, an autoimmune state may be induced by a pure dysregulation of the immune system. Such is the case in which severe systemic lupus erythematosus (SLE) is induced in young patients with myasthenia gravis following thymectomy. We have referred to this set of events as the 'kaleidoscope of autoimmunity'. Herewith, we would like to present another example of the kaleidoscope phenomenon, namely: the emergence of a full blown clinical presentation of the primary antiphospholipid syndrome (APS-recurrent thromboembolic phenomena, repeated fetal loss with high titers of anti cardiolipid antibodies) in a 32 y old female with myasthenia gravis, two years following thymectomy. Thymectomy in myasthenic patients may be associated with the emergence of new autoimmune conditions such as SLE and APS, pointing to the importance of immune dysregulation in the induction of these autoimmune diseases. PMID- 9229369 TI - Lupus erythematosus profundus around the salivary glands: a case resembling submandibular salivary gland disease. AB - In a 29 year old Japanese woman with SLE, a bilateral submandibular mass was detected. Computed tomographic (CT) scan suggested inflammatory changes around the submandibular glands. Histological findings of the deep skin biopsy showed typical changes of lupus erythematosus profundus (LEP). LEP should be considered in the differential diagnosis of an unexplained submandibular mass in a subject with SLE. PMID- 9229370 TI - Acinetobacter pericarditis with tamponade in a patient with systemic lupus erythematosus. AB - We describe a case of active systemic lupus erythematosus (SLE) complicated with a large amount of pericardial effusion with diastolic collapse of right ventricle suggestive of tamponade. Isolates from surgical drainage of pericardial fluid showed Acinetobacter baumannii exhibiting multiple antibiotics resistance. Despite the high frequency of both pericardial involvement and of infection complications in SLE, septic pericarditis and tamponade is considered rare. Most of the reported cases of septic pericarditis in SLE were due to Staphylococcal aureus, and Acinetobacter baumannii has never been reported before. PMID- 9229371 TI - Salivary testosterone during the menstrual cycle in women with systemic lupus erythematosus. PMID- 9229372 TI - Epilepsy and antiphospholipid antibodies in systemic lupus erythematosus patients. PMID- 9229373 TI - Transient neonatal bradycardia without heart block associated with anti-Ro antibodies. PMID- 9229374 TI - Protective immunity induced by vaccination with Onchocerca volvulus tropomyosin in rodents. AB - A cDNA clone of Onchocerca volvulus, designated MOv14, and encoding 136 amino acid residues from the C-terminus of O. volvulus tropomyosin, was evaluated as a protective immunogen in two complimentary rodent models of onchocerciasis. Vaccination of BALB/c mice with the recombinant fusion of MOv14 coupled to Maltose-Binding Protein (MBP) induced significant reductions (48-62%) in the recovery of Onchocerca lienalis microfilariae from the skin, compared to control groups immunized with MBP alone. The predominant antibody response generated to MOv14 by vaccination was of IgG1. Following a similar vaccination protocol in Mongolian jirds, two independent experiments demonstrated that 16 weeks after infection with Acanthocheilonema viteae there was a 46% reduction in the recovery of adult worms in vaccinated animals compared to control groups. Antibodies generated by vaccination recognized a product released during culture of A. viteae infective larvae which migrated at a distinct molecular mass from native tropomyosin from somatic tissues. PMID- 9229375 TI - Increased expression and secretion of ICAM-1 during experimental infection with Trypanosoma cruzi. AB - In the present study we demonstrate that spleens and hearts from BALB/c mice infected with the virulent Tulahuen or the low virulent CA-I strains of Trypanosoma cruzi, contain substantially higher ICAM-1 transcripts than uninfected controls. ICAM-1 expression in heart cells was also increased in the protein level, as measured by flow cytometry, ELISA and immunohistochemistry. The adhesive receptor was observed not only on inflammatory cells but also on sarcolemma of cardiac myocytes from T. cruzi infected mice. ICAM-1 expression was higher during the acute phase than in the chronic phase of infection, and paralleled the density of inflammatory leukocytes. Elevated titres of soluble ICAM-1 (s-ICAM-1) were detected in sera from mice during the acute phase of infection with CA-I or Tulahuen parasites. Cytokines, including IFN-gamma, IL-1 alpha, IL-6 and TNF-alpha have been shown to modulate expression of ICAM-1. Spleens and hearts from mice infected with CA-I or Tulahuen strains showed increased accumulation of mRNAs specific for these cytokines, which peaked during the acute phase of infection. However, IFN-gamma activity was not necessary for ICAM-1 induction, as its levels were also increased during infection in IFN-gamma receptor knock-out (IFN-gamma R- ) mice. Upregulation of ICAM-1 expression might be a direct consequence of parasite infection, since its density on cell lines of different lineages was enhanced after 24 or 48 h of infection with T. cruzi. PMID- 9229376 TI - Comparative vaccination of cattle against Boophilus microplus with recombinant antigen Bm86 alone or in combination with recombinant Bm91. AB - Cattle were vaccinated either with a single recombinant tick antigen, Bm86 or with a combination of two recombinant antigens, Bm86 and Bm91 from the tick Boophilus microplus. In three experiments, the responses of cattle to subsequent challenge with the tick were assessed. The addition of the Bm91 antigen enhanced the efficacy of the vaccination over that with Bm86 alone to a statistically significant degree. Moreover, co-vaccination with two antigens did not impair the response of cattle to the Bm86 antigen. Finally, responses of individual cattle to the two antigens were independent. All of these results may be relevant to the increase in efficacy expected from a dual antigen vaccine. PMID- 9229377 TI - In situ characterization of leucocyte subpopulations after infection with Eimeria tenella in chickens. AB - We characterized the leucocyte subpopulations after infection with Eimeria tenella in both naive and immune chickens. Immunocytochemical staining was used to characterize the cells in situ, so that the interaction between host and parasite could be studied. More leucocytes were detected in the lamina propria of immune chickens, and leucocytes infiltrated the ceca more rapidly than in naive chickens, but the infiltration was less pronounced than in naive chickens. In naive chickens, most infiltrated leucocytes were macrophages and T cells. Two days after inoculation the number of CD4+ cells had increased greatly. In immune chickens, mainly T cells (CD4+ and CD8+) infiltrated the lamina propria, and in contrast to naive chickens, the number of CD8+ cells exceeded the number of CD4+ cells. Furthermore, we characterized which cells contained a parasite and which cells were detected next to the parasites, because these cells are probably involved in the arrested development of the parasites. In naive chickens, sporozoites were significantly more often located within or next to macrophages than in immune chickens. In immune chickens, sporozoites were significantly more often located within or next to CD3+, CD8+, and TCR2+ cells. In conclusion, the marked increase of CD4+ cells after primary infection suggests that these cells are involved in the induction of the immune response, whereas the increase of CD8+ cells after challenge infection suggests that these cells act as effector cells. PMID- 9229378 TI - Comparative studies on immune responses to infection in susceptible B10.BR mice infected with different strains of the murine nematode parasite Trichuris muris. AB - A comparison of immune responses to infection between groups of B10.BR mice infected with different strains of T. muris, S strain (isolated in Sobreda, Portugal), E strain (isolated in Edinburgh), and E-J strain (originally E strain, which has been maintained in our laboratory, Japan), was performed. In mice infected with E and E-J strains, most of the worms were expelled by day 32 after infection, though the expulsion was faster in E-J strain-infected mice. In contrast, no expulsion was observed in S strain-infected mice by day 32 and egg production occurred on day 32. IL-4 production occurred in concanavalin A (Con-A) stimulated mesenteric lymph node cells (MLNC) from B10.BR mice infected with E and E-J strains, whereas no IL-4 production was observed in S strain-infected mice. IL-4 production did not occur in normal mice. In comparison with normal mice, high levels of IFN-gamma production by Con A-stimulated MLNC were detected in mice infected with every strain of T. muris. IFN-gamma production in S strain infected mice was greater, occurred earlier and was more persistent than in mice infected with E and E-J strains. IgG1 and IgG2a antibodies to T. muris excretory/ secretory antigens were observed in B10.BR mice infected with every strain of T. muris. Antibody production showed similar kinetics. These differences in the expulsion kinetics and IL-4 production in B10.BR mice infected with S, E, and E-J strains suggest the involvement of IL-4 in protection against T. muris infection, and confirm the previous conclusion by Else et al. PMID- 9229379 TI - Identification and characterization of human B-cell epitopes in recombinant antigens of Leishmania aethiopica. AB - Recombinant DNA fragments from Leishmania aethiopica that code for epitopes which react with human antibodies have been characterized by cross-hybridization studies and DNA sequence analysis. Twenty clones could be grouped into seven different groups (I-VII), probably representing seven different L. aethiopica antigens. The DNA sequences of representative clones from the seven groups have been obtained and the amino acid sequence of the respective recombinant antigens established. The recombinant antigens have been analysed by epitope scanning with patient sera, and octapeptides that contain potential B-cell epitopes have been identified in all seven recombinant antigens. These octapeptides have further been tested with additional patient sera and control sera, and three octapeptides (HAFCHEEG, YHSSVVHD and SYAPCSLK) were found to contain major epitopes recognizing specific antibodies in nine, seven and four, respectively, of the twenty sera tested. Fifteen of the twenty sera reacted with one or more of these three octapeptides. PMID- 9229381 TI - Reactive oxygen intermediates from eosinophils in mice infected with Hymenolepis nana. AB - A large number of eosinophils were recruited to the intestinal villi after infection with Hymenolepis nana. Eosinophil numbers were increased more rapidly in challenged mice than in primary infected mice. Local intestinal eosinophils from challenged mice showed more extracellular oxygen radical release, as assessed by histochemical methods using nitro blue tetrazolium, accompanied with tissue injury and larval degradation. Intestinal eosinophils isolated from the lamina propria induced specific oxygen radical generation in response to H. nana oncosphere extract as measured by luminol-dependent chemiluminescence. This response was stronger in challenged mice than in primary infected mice. Radical generation from uninfected mice was negligible. Lipid peroxidation in the small intestine, as measured by formation of malondialdehyde, was increased during H. nana challenge infection, the peak activity coinciding with the elimination of challenge larvae. Continuous administration of a NADPH oxidase inhibitor to sensitized mice interfered with the degeneration of challenge larvae. These results suggest that intestinal eosinophils may be the major contributor to oxygen radical production in response to H. nana and that reactive oxygen species may play a part of effector molecule in the resistance to reinfection with H. nana. PMID- 9229380 TI - Workshop on a detailed characterization of Trichinella spiralis antigens: a platform for future studies on antigens and antibodies to this parasite. AB - In order to characterize immunodominant components of T. spiralis a workshop was organized. In this the reactivity of monoclonal and polyclonal antibodies, provided by different research groups, towards total extracts from adult, new born larvae and muscle larvae as well as to excretory/secretory components of muscle larvae were tested by ELISA, Western blot and immunoprecipitation assays. As a result of this workshop T. spiralis ML antigens have been classified into eight groups (TSL-1-TSL-8) according to their recognition by monoclonal and polyclonal antibodies. Among them, TSL-1 antigens have been the most extensively characterized both biochemically and immunologically. These antigens are stage specific, originate in the muscle stichosome and are abundant in both E/S and on the larval cuticular surface. The TSL-1 antigens share an immunodominant carbohydrate epitope (tyvelose), which is unique for Trichinella and is not associated with phosphorylcholine. The data collected in this workshop has allowed both the unification of the nomenclature for T. spiralis antigens and their biochemical characterization. It also has provided a platform for further studies on the characterization of other T. spiralis antigens and indeed for other Trichinella species. PMID- 9229382 TI - Antibabesial effect of the immunomodulator AS101 in mice: role of increased production of nitric oxide. AB - The immunomodulator AS101 has been shown to induce cell proliferation and to increase the secretion of a variety of cytokines. In the present study we evaluated the effect of AS101 on the pathogenicity of B. rodhaini infected mice. In order to clarify its mechanism of action we studied the ability of AS101 to activate neutrophils and macrophages, both of which inhibit parasite growth. More specifically, we studied the ability of AS101 to induce secretion of nitric oxide (NO). We found that AS101 protects mice from babesiosis in a time and dose dependent manner. At 10 and 20 micrograms/injection, two weeks prior to parasites, AS101 significantly increased the number of neutrophils and more than doubled the survival rate of infected mice. Similarly, at these concentrations when injected one month, or at 20 micrograms, injected 24 h before parasites. AS101 mitigated the course of infection and reduced by half the peak of parasitaemia. At 0.1 microgram/ml AS101 induced the secretion of significantly higher levels of NO in vitro than control. This was abrogated by adding the NO synthase inhibitor. NG-monomethyl-L-arginine. In vivo the antiparasitic protection of AS101 was abrogated by another NO synthase inhibitor, aminoguanidine. We found that AS101, partly by elevating levels of NO, can significantly mitigate the course of infection and thus increase survival, and may therefore be proven as an effective antiparasitic drug. PMID- 9229383 TI - Heterologous expression of the cuticular-glutathione peroxidase of lymphatic filariae in an attenuated vaccine strain of Salmonella typhimurium abrogates H-2 restriction of specific antibody responses. AB - The major soluble cuticular glycoprotein of lymphatic filariae, gp29, has been expressed in the Salmonella typhimurium aroA aroD live vaccine strain BRD509. Two distinct constructs were generated: a) pgp29, in which gp29 was expressed directly via the inducible promoter nirB, or b) pTetC-gp29, in which it was expressed as a C-terminal fusion to the non-toxic immunogenic fragment C of tetanus toxin, again under the control of nirB. In both cases, plasmid stability in vivo was demonstrated by recovery of recombinant bacteria from livers and spleens of mice immunized via the intravenous route. Negligible gp29-specific antibodies were detected in animals immunized with bacteria expressing the fragment C fusion protein, but bacteria expressing the non-fused protein resulted in gp29-specific antibody production in a proportion of animals immunized. Notably, a number of BALB/c and B10.D2/n (i.e. mice of the H-2d haplotype) responded, in contrast to previously documented nonresponsiveness during infection, or immunization with parasite extracts. Presentation of gp29 by live attenuated S. typhimurium resulted in a broad spectrum of antigen-specific IgG isotypes. PMID- 9229384 TI - Neutralization of bovine concanavalin-A T cell supernatant-mediated anti-Cowdria ruminantium activity with antibodies specific to interferon gamma but not to tumor necrosis factor. AB - In an earlier study we demonstrated that Concanavalin-A stimulated bovine T cell supernatants inhibited the growth of Cowdria ruminantium in bovine endothelial cells in vitro. An investigation was conducted to identify the cytokines which were responsible for this growth inhibition. Addition of antiserum against bovine interferon gamma (IFN gamma) reproducibly neutralized the inhibitory effect of the T cell supernatants, whereas addition of antisera against bovine tumor necrosis factor alpha (TNF alpha) had no effect. The inhibitory effect of IFN gamma on C. ruminantium growth was not mediated by the production of nitric oxide as there was no detectable difference in nitric oxide levels in cultures that were supplemented with T cell supernatants compared with those that were not. The IFN gamma mediated anti-C. ruminantium effect highlights the importance of cell mediated immune responses in control of these infections and in particular incriminates the protective role of T cells, or cells that secrete IFN gamma. PMID- 9229385 TI - Protection of rats against malaria by a transplanted immune spleen. AB - A number of reports have suggested that the spleen plays a key role in the regulation of immunity to malaria but the role, if any, of other tissues is less clear. Furthermore, numerous functional changes occur in the spleen following malaria infection and it is not known whether the spleen's role relates primarily to its content of malaria-specific lymphocytes or to the altered structure and function that has occurred. To address these issues we have generated splenic chimeras by transplanting spleens between Plasmodium berghei-immune and naive rats. In the absence of a functional spleen, specific immune responses from both isolated splenic and non-splenic cells can partially control infection. However, an immune spleen in a naive rat can solidly protect the animal from malaria and a normal spleen in an otherwise immune rat can provide enhanced protection over the non-splenic state. Thus, in the presence of functional splenic architecture both splenic and non-splenic malaria-specific lymphocytes operate more effectively. However, these studies do demonstrate an important role for non-splenic tissue in immunity at least for P. berghei in the rat. The study could have important implications for induction of protective immune responses by vaccination and suggests that malaria-specific lymphocyte responses induced in the periphery following vaccination could interact with parasites in both spleen-dependent and spleen-independent ways. PMID- 9229386 TI - Selected P. falciparum specific immune responses are maintained in AIDS adults in Burkina Faso. AB - In tropical areas where Plasmodium falciparum malaria is endemic, co-infection with HIV-1 does not lead to a worsening of malaria, raising questions about the immunological interactions between both infections. Alterations of immune response to malaria during HIV-1 infection was investigated in the town of Bobo Dioulasso, Burkina Faso. Sixty-six adults were enrolled, including 37 HIV-1 positive subjects with < 250 CD4+ cells/microliter and clinical AIDS, and 29 HIV 1, negative healthy subjects. In vitro lymphocyte proliferation and cytokine (IFN gamma, IL-2 and IL-4) production were assessed in isolated mononuclear cells (PBMC) in presence of PHA, PPD or three malarial antigens: the baculovirus expressed protein from P. falciparum Merozoite Surface Protein-I, a P. falciparum in vitro culture and a crude schizont extract. Compared with healthy subjects. AIDS patients presented with decreased levels of cell proliferation and of IFN gamma and IL-2 production, in response to all antigens except the schizont extract. Similar levels of IL-4 production were obtained in both groups. Mitogenic stimulation of whole blood cultures was also performed, and revealed similar trends in cytokine production as in PBMC cultures. These results show that some components of the specific human immune responses to falciparum parasites may not be modified during AIDS, in spite of the strong cellular alterations induced by HIV, namely the decrease of the CD4+ lymphocyte subset. PMID- 9229387 TI - The IgG isotypes of specific antibodies in patients with American cutaneous leishmaniasis; relationship to the cell-mediated immune response. AB - American cutaneous leishmaniasis (ACL) presents a spectrum of clinical and immunological manifestations. Since the nature of the cellular response appears to play a fundamental role in determining the characteristics of the immunoglobulin isotype of specific antibody responses, we have compared the relative levels of specific antibodies of the four IgG isotypes against Leishmania in sera from patients with different clinical manifestations of ACL. Using a specific antibody capture assay, significant levels of antibodies of the IgG1, 2 and 3 isotypes were detected in localized cutaneous leishmaniasis (LCL); the average level of IgG4 antibodies was low and they were not detected in 10/20 sera. Sera from muco-cutaneous leishmaniasis (MCL) gave a comparatively strong IgG1 response. Sera from diffuse cutaneous leishmaniasis (DCL), the rare form characterized by antigen-specific anergy of cell-mediated immunity, showed highly significant levels of IgG4 antibodies compared to antibody levels of this isotype in the other groups; IgG1 and IgG2 levels were also elevated. Based on other studies of the relationship between the IgG isotype response and cell-mediated immunity, these results confirm a Th1-like CD4+ T cell response in LCL and MCL and a significant Th2-like response in DCL. PMID- 9229388 TI - Antigenic polypeptides of Echinococcus granulosus oncospheres and definition of protective molecules. AB - Immunoblotting and in vitro oncosphere-killing were used to identify a putative protective molecule in Echinococcus granulosus mature oncospheres. A range of sera from sheep that had been shown to be protected against E. granulosus, and from those that were not, were tested. The sera used were obtained from sheep hyperimmunized with E. granulosus oncospheres, or immunized with oncosphere non denatured extract, with immature oncosphere extract or with denatured extracts of oncospheres. Results indicated the involvement of native antigens of 23, 25, 30, 34 and 40 kDa in the protective response to E. granulosus infection. The rapid appearance of antibodies to the 23, 25 kDa antigens, their association with early onset of protection and in vitro oncosphere lysis by affinity-purified antibodies obtained from these fractions, indicated that these antigens contained protective epitopes. Final confirmation was provided by immunization of sheep with fractions prepared by preparative SDS/PAGE, and challenge infection. Only the fraction containing the 23 and 25 kDa molecules was able to stimulate protection. Antisera against this pair of molecules should provide a useful probe for screening an E. granulosus oncosphere cDNA library to identify clones expressing protective molecules. PMID- 9229389 TI - Kinetics and mechanism of effector focus formation in the lungs of mice vaccinated with irradiated cercariae of Schistosoma mansoni. AB - The sequence of events involved in effector focus formation around challenge schistosomula in the lungs of mice vaccinated with radiation-attenuated cercariae of Schistosoma mansoni has been characterized following intravenous administration of lung stage larvae. Histopathological analysis of the lungs of vaccinated animals revealed that infiltrating cells were present around larvae within 24 h. The main increment in cell recruitment occurred between 2 and 4 days, with foci reaching maximal diameter on day 8. No additional infiltration of the airways was detected by bronchoalveolar sampling before day 4 when the maximum number of cells, predominantly lymphocytes, was recovered. In contrast, responses in challenge control animals were relatively slight prior to day 12. IFN gamma was the major cytokine in airway cultures from vaccinated mice, the greatest increment in production coinciding with peak cell recruitment. A similar pattern of IFN gamma mRNA expression was observed in whole lung extracts, highlighting the dominance of Th1 responses in the effector mechanism. The slow start to focus formation may be due to the need for antigen, released by the intravascular parasite, to be translocated across the endothelium, processed by accessory cells and presented to the helper T cells which orchestrate the effector mechanism. The delay is of the same order as the period of development which the parasite must undergo in the lung, to facilitate further migration. This similarity in the timing may explain why some larvae are able to avoid the consequences of the pulmonary effector response. PMID- 9229390 TI - Comparison of complement activation in vitro by different Echinococcus granulosus extracts. AB - In the present study we have investigated and compared in vitro the specific complement (C) activating activity of three metacestode preparations of Echinococcus granulosus. Extracts from hydatid cyst fluid (HCF-ext), protoscoleces (PSC-ext) and hydatid cyst membrane (HCM-ext) activated human C producing C3 conversion and generating the C5b6 complex and the terminal C complex (TCC). HCM-ext showed much lower C activating activity than PSC-ext and HCF-ext. Moreover, its ability to generate C5b6 and TCC was lower than its ability to convert C3. On the other hand, PSC-ext and HCF-ext proved to be good C activators when their specific C activating activities were compared with that of inulin. However, PSC-ext produced lower levels of TCC than those produced by HCF ext, in spite of the fact that both produced practically the same levels of C3d and C5b6. These results may be consistent with the existence of several mechanisms of C modulation involved in the defence of the parasite against host C damage. PMID- 9229391 TI - Human isotype antibody responses to an Onchocerca volvulus glutathione S transferase. AB - Human isotype specific antibody responses to a recombinant pi-class glutathione S transferase (Ov24) from Onchocerca volvulus were assessed by ELISA, using a large and well-characterized bank sera (n = 238) from an hyper-endemic area of moderate intensity from Sierra Leone. IgG1, IgG4 and IgA responses, but neither IgG2 nor IgE response, to Ov24 were detected in infected subjects. The relationships between Ov24 antibody levels and skin microfilarial density, number of nodules, age, sex, eosinophil counts and clinical sign of reactive and chronic pathology were analysed using Pearson's correlation coefficient. Significant correlations between both IgA and IgG3 antibody levels and age were found (P < 0.01). Although no firm conclusions could be drawn from this study sample regarding the relationships between antibody levels and parasite load or clinical status, a negative correlation (P = 0.06) between Ov24 IgG3 antibody levels and microfilarodermia was found. PMID- 9229393 TI - Antibody response of Echinococcus granulosus infected mice: recognition of glucidic and peptidic epitopes and lack of avidity maturation. AB - The antibody response was followed weekly during 68 weeks in 17 Balb/c mice intraperitoneally (i.p.) infected with 2000 Echinococcus granulosus protoscoleces (PSC) and in three mice i.p. immunized with 2000 dead PSC. Antibodies against hydatid cyst fluid (HCFA) and its peptidic (periodate-resistant) and carbohydrate (periodate-sensitive) epitopes were titrated by ELISA. Avidity and the antigen recognition pattern of antibodies were also analysed during infection and immunization by ELISA and immunoblot, respectively. The antibody response of infected mice showed quantitative and qualitative variations during infection, since both titre as well as recognition of peptide and carbohydrate epitopes in HCFA depended on time post infection. No avidity maturation was evident during the course of infection. Sera from infected mice recognized the 38 kDa subunit of Ag5 but did not react with the 8 kDa subunit of AgB. On the contrary, the antibodies response of immunized mice showed only one peak of antibodies that recognized both peptidic and carbohydrate epitopes of HCFA. In addition, sera from these mice recognized mainly 60 and 110 kDa bands. Our results suggest that: a) avidity and antigen recognition patterns of antibodies in mice treated with live PSC are different from those treated with dead PSC; b) antibodies against HCFA glucidic or peptidic epitopes appear at different times post infection. PMID- 9229392 TI - Experimental model for human intestinal microsporidiosis in interferon gamma receptor knockout mice infected by Encephalitozoon intestinalis. AB - Interferon gamma receptor knockout mice developed a chronic infection when inoculated with spores of Encephalitozoon intestinalis which is a cause of intestinal microsporidiosis in AIDS patients. The infection was evaluated by enumeration of the spores of the parasite shed in the stools, histological examination and follow up over a period of six months. A dose-response was demonstrated since higher numbers of spores were excreted and more infection sites were found in mice which were given an increased quantity of parasites. In infected wild type mice, the number of excreted spores decreased until day 16 post-inoculation, then spores were detected sporadically in low numbers. These data confirmed the role of IFN-gamma in the control of E. intestinalis infection. The infection was not lethal suggesting that other factors are involved in regulation of the parasite infection. This model, with the long survival time of the animals together with the measurable quantity of spores shed in the stools will be useful for testing potential therapeutical agents. PMID- 9229394 TI - Homologous and heterologous protective immunity to Egyptian strains of Schistosoma mansoni and S. haematobium induced by ultraviolet-irradiated cercariae. AB - C57BL/6 and Balb/c mice were immunized with ultraviolet-irradiated cercariae of Egyptian strains of Schistosoma mansoni and S. haematobium, challenged with nonirradiated cercariae of the homologous or heterologous species, and assayed for protection against challenge infection by comparing the adult worm burdens of immunized and non-immunized mice. Homologous protection (per cent reduction in worm recovery) ranged from 56% to 69% for S. mansoni and 88% to 99% for S. haematobium. Significant heterologous protection was consistently induced against S. haematobium by immunization with S. mansoni, but not against S. mansoni by immunization with S. haematobium. These results are discussed in relation to those of previous studies and in terms of implications for vaccine development. PMID- 9229396 TI - A role for tumour necrosis factor-alpha in acute lymphatic filariasis. AB - A spectrum of clinical manifestations is a feature of human lymphatic filariasis. The acute disease is characterized by periodic and self limiting episodes of adenolymphangitis, fever and associated constitutional symptoms, while the chronic disease includes long lasting manifestations such as lymphoedema and/or hydrocoele. The microfilariae carriers are generally free of clinical symptoms. In the present study circulating Tumour Necrosis Factor (TNF-alpha) was measured in human bancroftian filariasis with different clinical manifestations. Significantly elevated levels were observed only in patients with acute disease and not in microfilariae carriers or in patients with chronic manifestations. A detailed analysis of the acute cases indicated an absence of correlation between TNF-alpha levels and duration of the episodes. However, a significant positive correlation was observed between the severity of the disease and the TNF-alpha levels. About 85% of the acute cases with severe manifestations showed raised levels of TNF-alpha while only 6.5% of mild cases showed such levels. Manifestation of fever was also significantly associated with higher levels of TNF-alpha-while 80% of acute cases with fever had significant levels only 24% of acute cases without fever had high levels of TNF-alpha. Based on these observations we propose a mediatory role for TNF-alpha in acute filariasis and the possible use of TNF-alpha inhibitors for clinical management of the disease. PMID- 9229395 TI - Differential antibody recognition of FC27-like Plasmodium falciparum merozoite surface protein MSP2 antigens which lack 12 amino acid repeats. AB - Three alleles of the FC27-type allelic family of the MSP2 gene of the malaria parasite Plasmodium falciparum have been sequenced from parasites from the field (The Gambia and Tanzania). These alleles lack the 12 amino acid repeat units which are usual in this family of MSP2 alleles. We have investigated the recognition by sera from an endemic area (The Gambia) of three recombinant MSP2 proteins that have 5, 1 and no copies of this repeat region. Antibody recognition of these recombinant proteins varied according to the number of repeats present. High titre antibody levels were seen with most sera using the recombinant protein with 5 x 12-mer repeats, whereas only low responses were measured using proteins containing 1 or no 12-mer repeats. Several sera entirely failed to recognise the protein which lacked 12-mer repeats. The data suggest that variation in the number of tandem repeat sequences could allow the parasite to avoid high avidity antibody binding and this may allow escape from immune recognition. PMID- 9229397 TI - Kinetics of nitric oxide production during infection and reinfection of mice with Plasmodium chabaudi. AB - We have shown previously that at the time of peak primary parasitaemia of P. chabaudi infection in NIH mice, significant levels of nitric oxide are produced, detectable as nitrate in the serum, and that these contribute to the protective immune response to infection. Here, we demonstrate that following reinfection, mice show a markedly diminished ability to produce nitrate. However, if mice are treated with L-NG-monomethyl arginine specifically to block nitric oxide metabolism during the primary infection, and are then reinfected, production of nitrate is restored to levels approaching those attained at peak primary parasitaemia. These experiments, together with others we have reported, indicate that whereas nitric oxide appears to play a significant role in control of the primary parasitaemia of P. chabaudi infection, it performs no such function during subsequent patent parasitaemias. Furthermore, they suggest that factors as yet unknown may regulate nitric oxide activity during malaria infection, such that under normal circumstances its production comes under strict control. This is exemplified by the observation that after the burst of nitric oxide activity that coincides with peak primary parasitaemia, there follows a prolonged period of immunological tolerance during which nitrate levels remain low even at secondary challenge infection. This tolerized state is lifted only several months after initial infection, when the nitric oxide activity at reinfection appears to correlate with the size of the parasite challenge and the presence of a patent parasitaemia. The implications of these findings for protective immunity to malaria, malarial immunosuppression, and immunoregulation in general, are discussed. PMID- 9229398 TI - Benefits of hypercholesterolemia treatment. PMID- 9229399 TI - Simvastatin in the secondary prevention of coronary heart disease. PMID- 9229400 TI - The 4S study: a critical review. PMID- 9229402 TI - Cholesterol-lowering drugs for primary prevention? The WOSCOP Study. PMID- 9229401 TI - The 4S study: a critical review. PMID- 9229403 TI - Cholesterol and coronary heart disease: new data from the WOSCOP Study. PMID- 9229404 TI - Effect of clozapine on dopamine-induced inhibition of prolactin release from cultured rat pituitary cells. AB - Although the atypical antipsychotic agent clozapine has little propensity to induce hyperprolactinemia in humans it increases serum prolactin levels in the rat. In this study, the effects of clozapine and of some typical antipsychotic drugs on basal and dopamine-mediated prolactin secretion from cultured rat pituitary cells were compared. Despite being less potent than the other antipsychotic agents tested, clozapine reverted the effect of dopamine on prolactin secretion in vitro. This finding suggests that clozapine interferes with dopamine receptors in the pituitary gland. PMID- 9229405 TI - Inhibition of steroid sex hormones release in rats by two Ca2+ channel blockers. AB - The calcium channel blockers Diltiazem HCl (2 mg kg-1 day-1, IP) or Cinnarizine (8 mg kg-1 day-1, IP) were given for a period of 30 days to adult male and female albino rats. The effect of each calcium channel blocker on circulating blood levels of steroid sex hormones was investigated by radioimmunoassay in comparison with the normal control level. The data demonstrated that both Diltiazem and Cinnarizine significantly decreased serum normal testosterone levels in males (36% and 52% inhibition respectively) as well as both normal estradiol and progesterone contents in females (58% and 45% inhibition with Diltiazem and 68% and 52% inhibition with Cinnarizine respectively). This study indicates the importance of blood sex hormones-follow up in case of long term Ca2+ channel blockers medication. PMID- 9229407 TI - Clearance, turnover time, and volume of distribution. AB - In mathematics a parameter is a fixed quantity that determines the behavior of a function. In the experimental sciences a parameter is an observable quantity that remains constant for every definable state of a system. In pharmacology it is important to distinguish among pharmacokinetic parameters, model parameters, and incidental parameters. A pharmacokinetic parameter is a quantity that defines a pharmacokinetic property of a biologic entity; conversely a model parameter defines the property of a chosen model, and an incidental parameter defines only the result of an experiment. The three most important pharmacokinetic parameters are clearance, turnover time, and volume of distribution. They are related by the expression. Clearance x Turnover time = Volume of distribution. PMID- 9229406 TI - Nicotine and stress: effect on sex hormones and lipid profile in female rats. AB - This work aimed to study the changes in sex hormones and lipid profile in adult female albino rats subjected to treatment with nicotine (N), immobilization stress (S), or their combinations (N+S). These treatments were applied either for one day (T1) or daily for 10 days (T10), after which rats in the estrus stage were used for the determination of plasma corticosterone (CS), serum sex hormones as progesterone (P), estrogen (E), FSH, LH and serum lipid profile including total cholesterol (TC), HDL-C, LDL-C, triacylglycerol (TG) and non esterified fatty acids (NEFA). It was clear that either N or S raised plasma CS and serum P levels in both the treatment regimens and that N+S induced a higher level of these hormones compared to each treatment alone. Serum E level was only elevated during T10 regimen only. An increase in serum LH level was only observed after a single exposure to either N or S, however their combination abolished the stimulatory effect induced by each treatment alone. Serum FSH was not altered by exposure to either N or S alone in both regimens, but in the T10 regimen their combination significantly lowered FSH level. Regarding the effect on serum lipid profile, serum TC was increased in all T10 regimen groups. LDL-C was increased by N+S treatment in both regimens, however no change in HDL-C level was observed in all groups. Serum NEFA was increased in all the treated groups during T10 regimen, while in the T1 regimen NEFA level was only elevated by the combination N+S. Serum TG was insignificantly altered in all the treated groups. The observed changes in the lipid pattern were attributed to the alterations occurred in CS and female sex hormones that caused by N, S or their combinations. PMID- 9229408 TI - Esophageal carcinoma: current controversies. AB - The incidence of esophageal adenocarcinoma and adenocarcinoma of the gastric cardia has increased so substantially in the last two decades that adenocarcinoma now accounts for approximately one half of esophageal malignancies seen in the United States and Europe. The reasons for this histological change may be related to a parallel increase in the incidence of gastroesophageal reflux disease in the Western world and the subsequent development of Barrett's metaplasia. Controversies surrounding carcinoma of the esophagus that are currently the focus of study are the relationship of Barrett's esophagus to the development of adenocarcinoma; whether adenocarcinoma of the esophagus and cardia is the same disease; the correct way to stage the disease; the treatment of disease confined to the mucosa; the extent of surgical resection to cure disease beyond the mucosa; the role of adjuvant chemotherapy in the treatment of the disease; and the methods of palliating patients with incurable disease. PMID- 9229409 TI - Intra-aortic ultrasonography in advanced esophageal cancer. AB - Thirty-one advanced esophageal cancer patients underwent preoperative intra aortic ultrasonography (IAUS), and aortic invasion was found in nine patients. In five patients, the aortic invasion was diagnosed as limited in the aortic adventitia, enabling the preoperative prediction of the tumors. The aortic invasion to all layers was visualized by IAUS in four cases. Intra-aortic ultrasonography provides important information to determine the resectability of advanced esophageal cancer. PMID- 9229410 TI - Ivor Lewis procedure for epidermoid carcinoma of the esophagus. A series of 264 patients. AB - Since 1982, Ivor Lewis esophagectomy has been performed on 264 patients with epidermoid esophageal carcinoma in our series. The mean age was 59. There were 243 men and 21 women; 91 patients had respiratory or cardiovascular problems. Two hundred forty-eight tumors were located in the lower two thirds of the esophagus. Sixty-eight patients had preoperative radiochemotherapy with cisplatinum and 5 flourouracil. One half of the resected specimens showed no residual tumor after radiochemotherapy. Tumor infiltration was T3 or deeper in 162 specimens, and 142 were N0. The main complications were respiratory (16%) and leaks (7%). Respiratory insufficiency was always fatal, but only 16% of the leaks led to death. The overall postoperative mortality was 4.5%, and the overall 5-year survival is 33.3%. Only T1 tumors have a significantly better prognosis (53.2% 5 year survival) as compared to T2 (30.6%) and T3 (27.2%), both at 5-year survival. Negative lymph node patients have a significantly improved 5-year survival rate of 44.8% vs. 15.2% for node-positive patients. For T3 tumors, preoperative radiochemotherapy seems to improve survival. Comparison of Ivor Lewis esophagectomy with other procedures shows no radical differences in complications. The 5-year survival rate seems unaffected by the procedure chosen; radiochemotherapy and extended lymphadenectomy still need further assessment. PMID- 9229411 TI - Barrett's cancer: indications, extent, and results of surgical resection. AB - In the Western world, the prevalence of Barrett's carcinoma, i.e., adenocarcinoma of the distal esophagus arising from specialized columnar epithelial metaplasia, has risen dramatically in the past two decades. High-grade dysplasia in the columnar epithelium has been identified as the precursor of malignant carcinoma. Whether an esophagectomy should be performed in patients with high-grade dysplasia remains controversial. Surgical resection is the mainstay of therapy in patients with invasive adenocarcinoma who are fit for surgery. Complete removal of the primary tumor and its lymphatic drainage has to be the primary goal of any surgical approach to adenocarcinoma of the distal esophagus. In patients with tumors located in the distal esophagus, this can be achieved by a radical transhiatal esophagectomy and proximal gastric resection with en bloc removal of the lymphatic drainage in the lower posterior mediastinum and along the celiac axis. Early adenocarcinoma can be cured by this approach. The value of multimodality therapy in patients with advanced tumors needs to be confirmed in well-designed randomized prospective trials. PMID- 9229413 TI - Thoracoscopic esophagectomy: are there benefits? AB - Between 1991 and 1995, 18 patients affected by a resectable intramural tumor of the esophagus underwent esophagectomy with thoracoscopic dissection of the esophagus. All patients had a relative contraindication to transthoracic esophagectomy with radical lymphadenectomy. All esophagectomies were completed thoracoscopically and reconstruction of the digestive tract was performed in 17 cases through cervical gastroplasty, and in 1 case, through cervical coloplasty. One cirrhotic patient died in the postoperative period due to a cervical anastomotic leak. Six other patients experienced a postoperative complication (mortality rate, 5.5%; morbidity rate, 33.3%). After a median follow-up of 17 months, 14 patients are alive without evidence of disease. One patient, who had excision of a cutaneous metastasis at a trocar insertion site 6 months postoperatively, eventually died with locoregional recurrence 14 months postoperatively. Another patient died 20 months after surgery with mediastinal recurrence. One patient died 28 months postoperatively after massive hematemesis with a suspect abdominal recurrence. The results of the present series, and those reported by other authors, do not seem to indicate evident advantages at present for the minimally invasive procedure during resection of the esophagus for cancer. Currently, there is no indication that this procedure should be used for standard clinical use. Wider randomized trials, performed in selected centers only, and longer follow-up are needed to further evaluate the procedure. PMID- 9229412 TI - Transhiatal esophagectomy for esophageal cancer. AB - The two main approaches currently used for surgical treatment of esophageal cancer are transhiatal esophagectomy (THE) and esophagectomy through a right thoracotomy. Among technical variations of THE, wide opening of the diaphragm with ample mediastinal exposure allows resection under direct view with acceptable postoperative morbidity and mortality rates. Transthoracic esophagectomy, associated with extensive mediastinal lymphadenectomy, still offers the best chance of definitive cure in intermediate stages (stages II and III), but does not influence survival in advanced cases (stage IV). In early stages, the lymph node invasion rate is negligible and may be treated by other techniques (THE or endoscopic mucosectomy). THE restores oral ingestion and avoids respiratory complications of thoracotomy, and consequently can be reserved for early cases (mucosal or submucosal lesions) or for patients with poor clinical status. To improve results of surgical treatment, protocols of associated radiochemotherapy are currently under research. PMID- 9229414 TI - Neoadjuvant chemotherapy with cisplatinum/5-fluorouracil/low-dose leucovorin for advanced squamous cell carcinoma of the esophagus. AB - Forty-four patients with advanced esophageal squamous cell carcinoma were treated with biochemical modulated combination chemotherapy and surgery. Treatment consisted of cisplatinum (70 mg/m2/day 1, day 22), 5-fluorouracil (5-FU; 700 mg/m2/day, days 1-5, 22-26), and leucovorin (20 mg/m2/day, days 1-5, 22-26) with nutritional support, and surgery (days 42-70, mean day 56). Surgery consisted of subtotal esophagectomy with extended lymphadenectomy. Postoperative adjuvant chemotherapy or additional irradiation to the mediastinum was restricted to patient with residual tumors. Clinical response rate was 63.6% in primary tumor, 52.6% in intramural metastasis, 100% in intraepithelial spread, and 30.9% for metastatic lymph nodes. There was a slight disagreement between the result of evaluation of histological and clinical effect. The incidence of postoperative complications was 25%, and the mortality rate was 2.3%. Overall 1-, 2-, 3-, and 4 year survival rates of the patients were 57%, 37.9%, 28.5%, and 28.5%, respectively. The median survival time was 14.7 months. Responders survived longer than nonresponders. The histological responders survived longer than clinical responders. The 4-year survival rate of patients without residual tumor after treatment was 75% in the superficial cases, 51% in the locoregional cases, and 50% in the widespread cases. PMID- 9229415 TI - Epidemiology and molecular biology of Barrett's adenocarcinoma. AB - In the United States, the incidence of esophageal adenocarcinoma has risen faster than any other malignancy in recent years, and now represents the most common histologic type of esophageal cancer observed in major institutions. The precise etiology of this malignancy, and the epidermiologic variables responsible for its dramatically rising incidence, remains obscure. Elucidation of the molecular biology of malignant transformation in Barrett's esophagus may improve the management of patients with advanced esophageal adenocarcinomas. Furthermore, appreciation of the molecular events associated with esophageal adenocarcinomas. Furthermore, appreciation of the molecular events associated with esophageal adenocarcinogenesis may facilitate early detection of occult carcinomas, and enable therapeutic interventions designed to prevent these otherwise highly lethal neoplasms. PMID- 9229416 TI - Esophageal cancer surgery: the value of controlled clinical trials. AB - Prospective randomized controlled trials (RCT) in esophageal cancer were reviewed. Their value and significance in the areas of multimodality therapy, lymphadenectomy, surgical techniques, palliative treatments, and perioperative management were evaluated. Much has been gained through RCT in the management of esophageal cancer, and RCT is the most reliable scientific method in clinical investigations. Problems in the conduct of such trials include lack of expertise by surgeons, lack of patients, lack of funding, and methodologic and ethical problems. These obstacles could be overcome and more well-conducted RCTs encouraged. PMID- 9229418 TI - Magnetic resonance imaging of osteonecrosis in divers: comparison with plain radiographs. AB - OBJECTIVE: To assess the diagnostic value of magnetic resonance imaging (MRI) as compared with radiographic findings in osteonecrosis in divers. DESIGN AND PATIENTS: MRI scans and conventional radiographs of the shoulder, hip and knee joints of 23 professional male scuba divers were reviewed together with their clinical findings and personal histories. Correlations between the MRI findings and the radiographic evaluation, clinical symptoms, and personal history were then investigated. RESULTS AND CONCLUSIONS: Lesions found on MRI in 23 divers included 27 in 39 proximal humeri, 17 in 36 proximal femora, 13 in 32 distal femora, and 12 in 32 proximal tibiae. Diffuse, marginated, or irregular patterns were observed. No lesions were seen in epiphyses of the distal femur or proximal tibia. We tried to classify these MRI findings by location and appearance. MRI showed no patients with only one affected bone. A close correlation between the MRI findings and maximum diving depth was observed in the proximal humerus. MRI depicted bone lesions that could not be detected on the radiographs. A routine MRI investigation of the hip joints should be performed in every diver in whom osteonecrosis is diagnosed at another site, for early detection of femoral head osteonecrosis. MRI of the shoulder joint is also the best surveillance in divers who dive deeper than 15 m. PMID- 9229417 TI - Benign and malignant cartilage tumors of bone and joint: their anatomic and theoretical basis with an emphasis on radiology, pathology and clinical biology. I. The intramedullary cartilage tumors. AB - We reviewed 845 cases of benign and 356 cases of malignant cartilaginous tumors from a total of 3067 primary bone tumors in our database. Benign cartilaginous lesions are unique because the epiphyseal plate has been implicated in the etiology of osteochondroma, enchondroma (single or multiple), periosteal chondromas and chondroblastoma. In the first part of this paper, we will review important clinical, radiologic and histologic features of intramedullary cartilaginous lesions in an attempt to support theories related to anatomic considerations and pathogenesis. PMID- 9229419 TI - Traumatic thoracolumbar facet instability: characteristic imaging findings. AB - OBJECTIVE: To review imaging patterns and injury mechanisms in patients with thoracolumbar facet instability (TFI). DESIGN: Imaging studies for thoracolumbar osseous injuries over a 2-year period were reviewed. Imaging findings, injury pattern and reported mechanism of injury were established for patients with TFI. PATIENTS: One hundred and ten patients with thoracolumbar acute, osseous injuries were studied. RESULTS: Eleven of 68 (16.2%) unstable thoracolumbar injuries demonstrated TFI. Seven (64%) of the eleven TFI patients were unrestrained occupants in a motor vehicle accident (MVA) and the remainder were involved in injuries dominated by blunt impact to the back. Only two (18%) had serious, permanent neurological deficits. CONCLUSION: TFI is a common injury pattern, particularly for unrestrained occupants in MVAs. Characteristic radiographic, CT and MRI findings are presented and correlated with the injury mechanism and clinical findings. PMID- 9229420 TI - Synovial chondrosarcoma arising in synovial chondromatosis of the right hip. AB - The case of a 55-year-old man with chondrosarcoma to the cervical spine is described. Two years previously the patient had undergone a right hip replacement for synovial chondromatosis. Re-evaluation of the biopsy specimen from the right hip taken at the time of the initial operation showed areas of chondrosarcoma arising in the background of synovial chondromatosis. The unusual presentation of this rare entity is discussed. PMID- 9229421 TI - Dedifferentiated chondrosarcoma in patients with multiple osteochondromatosis: report of a case and review of the literature. AB - Multiple osteochondromatosis (MOS) is a familial disorder of autosomal dominant transmission characterized by the development of multiple exostoses and often derangements of epiphyseal cartilage, sometimes resulting in long bone growth retardation. Patients with the disorder appear to be at increased risk for developing secondary chondrosarcomas. Rarely, dedifferentiated chondrosarcomas may also occur. We report a single case of a 27-year-old man with multiple osteochondromatosis who developed a fatal dedifferentiated chondrosarcoma. Radiographically, the neoplasm arose from the pelvis completely destroying the left pubic ramus. Subsequently, the patient underwent preoperative chemotherapy followed by a left external hemipelvectomy. On pathologic examination, the tumor was characterized by high-grade pleomorphic sarcoma sharply juxtaposed to a low grade chondrosarcoma. The patient ultimately died of widespread metastatic sarcoma. PMID- 9229422 TI - Periosteal chondrosarcoma invading the medullary cavity. AB - We present a case of periosteal chondrosarcoma of the femur, in which a tumor invaded the medullary cavity and the lesion was clearly demonstrated only on MRI. To the best of our knowledge, there have been no previous reports of an intramedullary lesion caused by periosteal chondrosarcoma demonstrated on MRI. PMID- 9229423 TI - Skip metastases in Ewing's sarcoma: a report of three cases. AB - The accurate pre-operative staging of all potentially malignant tumours of bone is essential. We report three cases of Ewing's sarcoma of bone in which MR imaging identified skip metastases not visualized on either contemporary radiographs or bone scintigraphy. The implications for patient management and possible reasons for the other imaging modalities failing to reveal the skip metastases are discussed. PMID- 9229424 TI - How should the Stroop interference effect be measured? Further evidence from alternative versions of the Stroop task. AB - The present study modeled the relationship between the Stroop and the neutral naming times to investigate the mechanism underlying the Stroop interference effect. 95 subjects took six alternative versions of the Stroop task and the naming times in the Stroop and neutral conditions were each averaged across the tasks to arrive at a more general measure of the Stroop and neutral naming times. Regression analysis of these general measures indicated a linear function with a small and nonsignificant intercept and a slope significantly greater than one. This finding is consistent with Chen's 1996 results and supports the hypothesis that the mechanism underlying the Stroop interference is interactive or multiplicative rather than stage-like or additive and that a ratio of Stroop over neutral naming times was psychologically a more appropriate measure. PMID- 9229425 TI - Scores on the Group Embedded Figures Test by undergraduates in information management. AB - The Group Embedded Figures Test was administered to 63 undergraduates in a program for a Bachelor of Applied Arts in Information management. Distribution characteristics, sex differences, reliability, and internal consistency measures for this sample were compared with those for Witkin's original sample. In addition, item difficulty and discrimination coefficients are provided. Scores for this group show desirable measurement characteristics. PMID- 9229426 TI - Exploring quality of life of adults with spinal cord injuries. AB - Measurements of quality of life have been an important research focus in rehabilitation and medicine. Analyses indicated that for 12 persons with spinal cord injury significant quality of life domains identified through a ranking procedure were different from domains identified through small group discussions. If replicated with larger groups, we would advocate direct responses obtained through small group or individual discussions. PMID- 9229427 TI - Consistency in the expression of aesthetic preference in line partitioning. AB - In experimental enquiries into the aesthetic properties of the golden section, investigators have tended to employ tests which either require the subjects to generate an artifact or require the subjects to choose between artifacts shown to them. These researchers have assumed that the findings from each type of test are equally valid. This assumption was investigated and shown to be tenable for the partitioning of line segments. Experimental subjects (N = 57) expressed an aesthetic preference for the same line partitioning ratios irrespective of the type of test employed. PMID- 9229428 TI - State-dependent psychiatric phenomena on the Rorschach: two case studies of acutely unstable suicidal mental status. AB - Two inpatient case studies are presented illustrating the clinical utility of the Rorschach to display and evaluate acute state-dependent psychiatric phenomena. The patients interviewed experienced severely acute, intrusive, ego-alien suicidal ideation. Comparison of test and retest data indicates correspondence between symptoms and test signs. Data suggest that Rorschach responses, given with serious and perhaps ominous affect involving inanimate movement, morbid content directly imputable to suicidal themes, and some co-occurrence of morbid content and inanimate movement, were closely associated with acutely unstable suicidal mental status and display remission coincident with clinical improvement. These efficiently available Rorschach data may be of value in substantiating acute mental status. PMID- 9229429 TI - Type II errors in comparisons of dextral and sinistral groups. AB - Anomalous cerebral dominance can have multiple manifestations, e.g., left-eye preference or left-foot preference in the absence of left handedness per se. The 1987 estimate of prevalence of anomalous dominance in the population by Geschwind and Behan approaches 30%, three times the estimate obtained by using handedness alone as the sorting criterion. Accordingly, substantial numbers of subjects assigned to "dextral" groups may actually display other legitimate indicators of anomalous dominance. As such there is a tendency to commit Type II errors in studies in which handedness alone is used to represent the variance attributed to cerebral dominance. Accordingly, null hypotheses assessing the relations of cerebral dominance with dependent variables are accepted even though they may, in fact, be false. PMID- 9229430 TI - Physical performance in relation to age, sex, social class and sports activities in kindergarten and elementary school. AB - Physical performance of preschool children and elementary school pupils (N = 2309, age: 61 to 108 mo.) was related to characteristics of physical growth and cognitive performance and to ecological variables. Correlations between measures of physical growth and physical performance and between physical and cognitive performance were positive and significant. Measurements of physical fitness and body coordination increased across ages. Significant differences were found between boys and girls; however, boys exceeded on some items, girls on others. Children of higher socioeconomic status performed better than children of lower status and children who participated in sports outside school outperformed those who did not. PMID- 9229431 TI - Ebbinghaus illusion: effect of figural similarity upon magnitude of illusion when context elements are equal in perceived size. AB - The magnitude of the Ebbinghaus illusion has been reported to be greater when test element and context elements are figurally similar as opposed to figurally dissimilar. In the current investigation with 16 observers, illusion magnitude was greater for a figurally similar configuration even though the context elements of the figurally similar configuration were perceived as smaller than the context elements of a figurally dissimilar configuration. Hence, figural similarity appears to have a prepotent effect in the Ebbinghaus illusion. PMID- 9229432 TI - Measuring chronic pain in children, an exploration. AB - The study focused on the feasibility and validity of pain instruments and the optimal period of diary registration for measuring chronic pain intensity of 13 children. Highly positive associations were found between the registration of pain on a Visual Analogue Scale and on the Postoperative Pain Measure for Parents. For children under medical treatment for chronic limb pain a one-week dairy registration suffices. PMID- 9229433 TI - Listening to Mozart does not enhance backwards digit span performance. AB - Rauscher, Shaw, and Ky recently reported that exposure to brief periods of music by Mozart produced a temporary increase in performance on tasks taken from the Stanford-Binet Intelligence Scale-IV. The present study examined whether this effect occurred in performance on a backwards digit span task. In a within subjects design 36 undergraduates were exposed to 10-min. periods of Mozart music, a recording of rain, or silence. After each stimulus period, undergraduates had three attempts to hear and recall different 9-digit strings in reverse order. No significant differences among treatment conditions were found. There was a significant effect of practice. Results are discussed in terms of the need to isolate the conditions responsible for production of the Mozart effect. PMID- 9229434 TI - Use of the Million Clinical Multiaxial Inventory in clinical practice. AB - A review of recent survey data indicates that the Million Clinical Multiaxial Inventory (MCMI) ranks among the most frequently used tests by practicing clinicians. This instrument is surpassed only by the MMPI or MMPI-2 in the area of objective personality assessment. PMID- 9229435 TI - Aging and executive function skills: an examination of a community-dwelling older adult population. AB - The purpose of the present study was to employ the Tower of Hanoi task to the study of possible changes in executive function skills in older adults. The study used a quasi-experimental design, with age group (i.e., young adult, young elderly, or older elderly), being the independent variable in examining performance differences between younger and older adults. Data were analyzed cross-sectionally by age group. Nineteen elderly men and women comprised two groups; nine Young Elderly with an average age of 65 years and ten Older Elderly with an average age of 75 years. Two men and ten women served as a Young Adult comparison group having an average age of 19 years. Performance on the Tower of Hanoi was measured by efficiency scores (number of trials to consecutive solutions), frequency of error types, self-correction scores (completing the goal configuration in twenty or fewer moves after committing an error precluding a "correct" solution), and error perseveration (committing the same error on two consecutive trials of a problem). Analysis of variance and chi-squared tests suggested similar executive capacities among the 9 young adult and the 8 young elderly participants as compared to their 7 older elderly peers on the 3-disk task. However, on the 4-disk task where problem complexity increased by the addition of another disk and longer move sequences, young adult participants showed superior performance on the average than either young elderly or older elderly participants. Although the present study is limited by the small sample size and the use of cross-sectional analyses to examine age differences, these findings are consistent with the hypothesis of age differences in executive function. PMID- 9229436 TI - Relationship of swimming distance, expectancy, and performance to mood states of competitive athletes. AB - This study focused on the relationship between normal and abbreviated training sessions for young competitive swimmers and acute changes in mood. Several potential moderators of the relationship between exercise and mood also were examined. 25 girls and 23 boys, swimmers between the ages of 12 and 25 years, completed a shortened version of the Profile of Mood States before and after normal-distance and taper practices. An hypothesized interaction between distance training and acute changes in scores on Total Mood Disturbances was significant. During normal-distance practices, scores on Mood Disturbance increased from pre- to postpractice. Analyses of the individual subscales indicated that swimmers" scores increased for Fatigue and decreased for Vigor. In abbreviated practice sessions, athlete's scores on Total mood Disturbance showed no change from pre- to postpractice. The specific subscales, however, showed positive changes for Depression, Confusion, and Tension. The mood changes related to practice distance were not influenced by the possible moderating factors of expectancy or performance times. Thus, even for highly trained competitive swimmers, exercising at or near maximal physical capability is associated with few positive changes in mood scores. Shorter-distance swims that do not tax endurance are preferable, if mood enhancement is a goal. PMID- 9229437 TI - Psychomotor and cognitive performance in nonapneic snorers: preliminary findings. AB - Snoring is a common phenomenon and a primary symptom in obstructive sleep apnea syndrome, a sleep-related breathing disorder in which neuropsychological function is reported to be impaired. The first purpose of the present study was to compare cognitive and motor function in 25 heavy nonapneic snorers and 26 sleep apneics. As the basis for impairments in heavy nonapneic snorers is still unclear, the influence of nighttime breathing disturbances and morning alertness, respectively, on daytime performance was evaluated too. Nonapneic snorers exhibit more slow wave sleep and tend to have fewer changes in sleep stage than sleep apnea patients, but values for other sleep variables are similar. Snorers also show comparable alertness. Deficits in immediate visual memory and in visuospatial reasoning are not found. However, there are some indications that snorers show decreased manual dexterity and eye-hand coordination for the nonpreferred hand and that they have deficits in focused attention. In addition, snorers may show difficulties in finger-tapping speed. These performance measures tend to be associated with reduced morning alertness, except for the score on focused attention which has a tendency to be related to the nocturnal breathing disturbances. PMID- 9229438 TI - Psychology of computer use: XLVI. Age-related differences in the mapping of auditory icons to visual icons in computer interfaces for children. AB - An investigation was conducted to characterize how children ages 6 through 9 identify auditory icons present in educational software. 24 subjects were require to map auditory icons to visual icons, both present in a computer interface. The interface used in the experiment was constructed with Visual Basic and involved 40 auditory icons, 40 corresponding visual icons, and 66 extraneous visual icons. It was hypothesized that older children would be better able to map the auditory icons to visual icons due to more extensive exposure to everyday sounds. The results supported the hypothesis and suggestions for additional research were provided. PMID- 9229439 TI - Comment on autogenic training and hypertension. AB - We comment on a report by Watanabe, et al. regarding the effects of autogenic training on hypertension. Using previous reports in the United States, we mention methodological problems on how to evaluate the effects of autogenic training and express our hope that they would provide further research to clarify the effects of autogenic training on hypertension. PMID- 9229440 TI - Adaptation to optokinetic rotation-induced motion sickness without experiencing nausea. AB - 17 subjects highly susceptible to motion sickness were divided into Nausea and No nausea groups depending upon whether the subjects experienced nausea during adaptation to nauseagenic rotation. Eight subjects in the Nausea group viewed an optokinetic rotation drum for repeated sessions of 16 min., with an interval of two days between every two sessions. During each session the subjects continuously viewed the rotating drum 16 min. even if they experienced nausea. nine subjects in the No-nausea group likewise viewed an optokinetic rotation drum for at most 16 min. but immediately stopped viewing the rotating drum whenever they experienced nausea during the session. The criteria for full adaptation were that the subject reported no feelings of stomach discomfort and nausea during a 16-min session. The mean number of sessions to reach full adaptation was 3.9 (SD = 0.6) with a range 3 to 5 sessions for the subjects in the Nausea group and 3.2 (SD = 0.8) with a range 2 to 4 for the subjects in the No-nausea group. It was concluded that subjects can adapt to a nauseagenic optokinetic rotation by repeated exposure without experiencing nausea and vomiting during adaptation. PMID- 9229441 TI - Hypoglycemia as a mitigating factor in vehicular accidents. AB - Cognitive, perceptual, and motor deficits are part of the constellation of symptoms found in various hypoglycemic conditions. Since neurogenic symptoms and altered states of consciousness affect driving skills, hypoglycemia should be considered a mitigating factor in vehicular accidents. PMID- 9229442 TI - Menstrual phase, history of smoking, and taste discrimination in young women. AB - University women who were either nonsmokers or who had smoked regularly for at least two years (ns = 7) attempted to discriminate a gustatory stimulus five days before and five days after their menses. The stimuli were presented 3 min. or 5 min after the introduction of a target stimulus. A statistically significant interaction was found among history of smoking, menstrual phase, and latency for taste comparisons. However, when the shared variance between olfactory and gustatory accuracy was partialled out, the results for smokers suggested significant increases in gustatory thresholds relative to those of nonsmokers regardless of menstrual phase. Smokers also displayed significantly less (eta 2 = 25%) accuracy for olfactory discrimination than nonsmokers. PMID- 9229443 TI - Time headway in car following and operational performance during unexpected braking. AB - The relation between car-following behaviour and braking performance was studied in a driving simulator. The theoretical perspective was that individual differences in tactical car-driving behaviour may be related to skills on the operational level of the driving task via a process of adaptation. In a sample of 16 young and middle-aged experienced drivers independent assessments were made of preferred time headway during car following and of braking skill. Starting from modern theories of visual-motor learning, braking performance was analyzed in terms of a reaction time component, an open-loop visual-motor component, and a closed-loop visual-motor component involving the precise adjustment of braking (timing and force) to the situation. The efficiency of the visual-motor component of braking was a strong and significant predictor of choice of time headway to the lead vehicle in such a way that less efficient braking indicated a preference for a longer time headway. This result supports the theory of adaptation on the individual level. PMID- 9229444 TI - Job satisfaction in casino employees. PMID- 9229445 TI - Children's lateralization on a finger-localization task. AB - The effect of stimulus orientation and hand use on performance on a finger localization task was assessed in a sample of 40 children, including 20 boys and 20 girls. Experimenters stimulated a single finger on either the left or right hand, and the participants were required to identify the stimulated finger displayed on a drawing of a hand. The model drawing was displayed in either an isomorphic or reversed orientation relative to the stimulated hand. Analysis indicated that 4-yr.-olds performed significantly better than 3-yr.-olds. Also, participants were significantly better at identifying the middle finger when the left hand was stimulated than the right hand. PMID- 9229446 TI - 3-dimensional kinematics of overarm throwing action of children age 15 to 30 months. AB - 7 children 15 to 30 mo. old participated in a study of 3-dimensional kinematics of overarm throwing. Children of different ages were considered to be at different developmental stages of motor development. Video recordings were digitised and 3-dimensional coordinates established using the DLT algorithm. Qualitative analysis indicated that the children executed either a 'static' or 'dynamic' throwing action. Either could further be classified as 'arm dominated' or 'sequentially linked.' Maximum elbow extension was no more than 163 degrees for any child; release velocity was higher for older subjects; and the angle of ball release was large in 'arm-dominated throws' (M = 49 degrees) and comparatively smaller in 'sequentially linked' throws (M = 15 degrees). PMID- 9229447 TI - Depression following brain trauma is enhanced in patients with mild discrepancies between intelligence and impairment on neuropsychological scores. AB - Analysis of the MMPI (Minnesota Multiphasic Personality Inventory) scores from 135 (20 years to 60 years old) patients who had sustained closed head injuries supported the hypothesis of a nonlinear relationship between the severity of depression and the magnitude of the discrepancy between intelligence and neuropsychological proficiency. Although the MMPI Depression T scores for all groups of patients were elevated (M = 78, SD = 13), patients with the least and greatest discrepancies between intelligence and neuropsychological proficiency scored lower on Depression than patients with discrepancies within the z-score ranges -2.0 and -1.1. The results of symmetrical covariance for either depression or complex partial epileptic-like experiences before comparisons between groups suggested depression and the epileptic-like experiences share the same source of variance. PMID- 9229448 TI - Olympic athletes' transition from sport to workplace. AB - Research has supported the need for strategies to assist elite athletes with transition from full-time athletic activity to the work place. Early intervention with coaches' and peers' support programs have mediated the problems associated with the termination of athletic careers. The present study is a report about 57 prominent Olympians from 12 disciplines spanning 6 0 years of competitive sports. Analysis of the data suggests that focussed efforts early in the athletes' careers assisted preparation for life after the full athletic activity. Mentors not only assist in the athletes' careers but also offer guidance and counsel during later life. PMID- 9229449 TI - Changes of immunoregulatory cells associated with psychological stress and humor. AB - For 8 medical students influences of psychological stress or humor on T-cell subset percentages and natural killer cell activity were investigated by measuring these parameters before and after an examination and before and after watching a comedy video. Although T-cell subsets were not significantly affected by either stimulus and natural killer cell activity was not affected by the examination, the latter was significantly decreased after watching the comedy video. PMID- 9229450 TI - Future time perspective and positive health practices in young adults: an extension. AB - A sample of 69 young adults attending a public university responded to the Future Time Perspective Inventory, two subscales of the Time Experience Scales (Fast and Slow Tempo), and the Personal Lifestyle Questionnaire in classroom settings. A statistically significant correlation (.52) was found between scores for future time perspective and the ratings for the practice of positive health behaviors in young adults. This correlation was larger than those previously found for middle and late adolescents. Scores on subscales of individual health practices and future time perspective indicated statistically significant correlations for five (.25 to .56) of the six subscales. Scores on neither Fast nor Slow Tempo were related to ratings of positive health practices or ratings on subscales measuring positive health practices. PMID- 9229451 TI - Autogenic training and dream recall. AB - The present study has investigated the relationship between Autogenic Training and dream recall for 112 participants in 16 beginning courses of 10 wk. Analyses confirmed the hypothesis that learning and practicing this relaxation technique enhanced dream recall. PMID- 9229452 TI - Background instrumental music and serial recall. AB - Although speech and vocal music are consistently shown to impair serial recall for visually presented items, instrumental music does not always produce a significant disruption. This study investigated the features of instrumental music that would modulate the disruption in serial recall. 24 students were presented sequences of nine digits and required to recall the digits in order of presentation. Instrumental music as played either forward or backward during the task. Forward music caused significantly more disruption than did silence, whereas the reversed music did not. Some higher-order factor may be at work in the effect of background music on serial recall. PMID- 9229453 TI - Differences in shot-making skills among high and low money winners on the PGA tour. AB - Analysis of individual performance statistics for the 1995 Professional Golf Association tour yielded statistically significant differences between the shot making skills of the to ten and bottom ten money winners. Only two skills (driving distance and total driving) were significantly higher among the ten top ranked money winners. PMID- 9229455 TI - Correlations of body composition and body-image assessments of adolescents. PMID- 9229454 TI - Adults and children with high imagery show more pronounced perceptual priming effect. AB - 36 children in Grade 5 and 59 university students, all native speakers of Japanese, studied three types of priming stimuli in a mixed list: words written in hiragana (Japanese syllabary used in writing), words written in kanji (Chinese characters also used in writing), and pictures. They were then given a task involving completion of hiragana-word fragments: the task involved studied and nonstudied items. For both children and university students, words in hiragana produced the largest priming effects, that is, the words that had appeared in hiragana in the preceding study phase were generated more often in the test phase of word completion than the other two types of priming stimuli. This confirms that the perceptual priming effect depends much on data-driven processing. For both age groups, words in kanji produced nearly half the priming effects seen for hiragana-words. On the other hand, pictures had no priming effect for children but they had a similar effect to kanji-words for students. The discrepancy between kanji-words and pictures for children suggests that the former force the subject to read the words, which, possibly, activates the hiragana-words, while the latter do not necessarily force labelling the pictures. Among three kinds of imagery tests, the Verbalizer-Visualizer Questionnaire predicted priming scores for children and the Questionnaire upon Mental Imagery did so for students, but the Test of Visual Imagery Control did not predict the scores for either age group. This shows that children reporting habitual use of imagery and adults reporting vivid imagery have more pronounced perceptual priming effects. We conclude that the imagery ability based on self-judgments reflects real characteristics of the perceptual representation system of Tulving and Schacter (1990). PMID- 9229456 TI - Perception of body size and body satisfaction in recovered anorexic women: comparison with restrained and unrestrained eaters. AB - The perception of body size, measured by three different methods, and body satisfaction were assessed in 23 formerly anorexic inpatients with an "intermediate" (n = 9) or a "good" outcome (n = 14) and compared with the data obtained from 21 restrained and 20 unrestrained eaters. Using the Kinaesthetic Size Estimation Apparatus, overestimation and uncertainty in the perception of body size became apparent in both groups of former patients. The other two methods, Video Distortion Technique and Image Marking Procedure, did not produce comparable results. There was only a trend towards higher scores on body dissatisfaction, as measured by the Body Shape Questionnaire, in the patients' groups in comparison with the group of unrestrained eaters, whereas the patients' scores on body dissatisfaction were quite similar to those of the restrained eaters. None of these measures discriminated between the two outcome categories of "intermediate" and "good.". These findings suggest that restoration of body weight, by itself, obviously does not cause a normalization of body experience in all its components in patients with anorexia nervosa. PMID- 9229457 TI - Effect of speaking into a passive resonator on stuttering frequency. AB - The effect on stuttering frequency of speaking into a passive resonator was investigated. Eight participants who stuttered read aloud with and without the benefit of the resonator. A statistically significant reduction of approximately 30% in stuttering frequency was observed while the participants spoke with the resonator. These and similar commercially available devices may be employed with individuals who stutter, particularly children, as a means of enhancing fluency. PMID- 9229458 TI - Color preferences of borderline patients and of normal controls. AB - 20 female patients who met the DSM-III-R criteria for the diagnosis of borderline personality disorder and 23 normal female controls were asked to rank-order eight Luscher color cards, at first with no specific use for the colors mentioned, then as a color for their own dress or jacket, then for their living room, and then as a color they would like their friend to wear. Very few statistically significant differences (p < .01, 2-tailed) between the groups were found. Borderline patients ranked more favourably than controls the use of black color for their living room (Pearson r = .51) and were less likely to favour grey color for this purpose (r = -.45). When no specific use for colors was mentioned, the borderline patients ranked red more favourably than the control group (r = .47). Within the normal control group, statistically significant differences between the ranks of the same color were noted depending on the particular use of the particular color. PMID- 9229459 TI - Detail, pressure, and completion of Draw-A-Person produced during silence or rock music. PMID- 9229460 TI - Trail Making Test in assessing children with reading disabilities: a test of executive functions or content information. AB - The speed of performance on Part A, Part B, and on an experimental version containing alphabetical series (Part A Alphabetic) of the Trail Making Test was studied with 19 children with reading disabilities and 34 controls from Grades 4 to 6. When the test was used in discriminant profile fashion, children with reading disabilities showed a deficit compared with control children on Part B relative to part A but did not relative to the new Part A Alphabetic. The results indicate that the performance of the children with reading disabilities on Part B is likely to be affected by their slowness on the alphabetical series. Based on these results we recommend that the speed of following the alphabetical series be assessed when using Part B of the Trail Making Test. PMID- 9229461 TI - Experience with perceptual and motor skills in rhythmic gymnastics. AB - Based on the notion of measuring motor performance, an experiment with three groups of 20 elite rhythmic gymnasts (N = 60), 9 to 10 yr., 11 to 12 yr., and 13 to 15 years of age (national level), with children of the same size and age was conducted, to identify the important abilities for the achievement of excellence in this sport. Motor abilities (whole-body coordination, dynamic balance, static balance, sense of kinesthesis, whole-body movement time, and eye-hand coordination) as well as perceptual abilities (whole-body reaction time, anticipation of coincidence, and depth perception) were compared. Analysis showed that scores on measures of whole-body coordination, dynamic balance, and static balance were higher for elite groups of athletes than for corresponding control groups. Moreover, elite athletes in the oldest group scored higher than those in the youngest group on anticipation of coincidence, on eye-hand coordination, and on static balance. These findings indicate the presence of systematic differences between elite athletes and nonathletes on motor abilities related to experience in this sport. PMID- 9229463 TI - Role of pictorial cues in young children's distinguishing self-performed from self-imagined actions. AB - 32 4- and 5-yr.-olds participated in a series of performed and imagined actions and a memory interview. Children in the Picture condition answered questions accompanied by pictures of actions whereas children in the Verbal condition received only the verbal cues. Children in the Picture condition performed as well as children in the Verbal condition when classifying performed and new actions but had more difficulty classifying imagined actions. Results suggest that retrieval cues (pictures) did not enhance children's discrimination of self performed and self-imagined actions. PMID- 9229462 TI - Stability of the Professional Development Assessment. AB - The Professional Development Assessment was constructed and pilot-tested with 76 students in three occupational therapy programs. A comparison of pretest and posttest scores yielded a significant correlation of .48, supporting the stability of responding over 1 to 2 years and suggesting usefulness of further development for evaluation of professional behaviors in students. PMID- 9229464 TI - Effects of duration of stimulus and variability of foreperiod on the identification of multidimensional stimuli. AB - Two experiments were conducted to investigate the effects of duration of stimulus and variability of foreperiod on the identification of multidimensional stimuli. Statistical analysis showed that performance speed and accuracy deteriorated as duration of stimulus was severely limited. Further, subjects seemed to change the allocation of attentional resources according to the attributes of stimulus. They tended to distribute more attentional resources to the less salient attribute which resulted in a statistically nonsignificant effect of order of report under time stress. Variability of foreperiod had very little effect on performance and may not be important to consider in reactions. Implications of these results for the design of multidimensional displays and for human information processing were discussed. PMID- 9229465 TI - Correlation between field dependence-independence and handball shooting by Swedish national male handball players. AB - Contradictory claims exist as to whether field dependence or field independence is advantageous to team ball-game performance. For further investigation, Swedish national male handball players' Rod-and-Frame Test scores were correlated with their field-goal shooting attempts and shooting efficiency in the '94 European Handball Championship. No significant correlation was found; discussion followed. PMID- 9229466 TI - Effects of speed and different types of treadmill on the range of motion of the lower extremities. AB - This study examined the range of motion of the lower extremities in 20 subjects (10 men, 10 women) exercising on treadmills. Of particular interest was how this parameter was affected by speed and different types of treadmill. Analysis indicated that there were statistically significant differences in the hip and knee joints, but not in the ankle joint for two selected types of treadmill set at the same speed. PMID- 9229467 TI - Factors related to perinatal substance abuse in a California county. AB - The study reported here originates from a county in California between Los Angeles and San Francisco. 658 mothers were tested over a 1-mo. period in June 1991 to ascertain the prevalence of drug misuse among pregnant women. 11% of the mothers tested positive for some type of substance. Of those substances found in the urine of the mothers who tested positive, the most prevalent were barbiturates, marijuana, methamphetamines, and amphetamines. When the multivariate analysis of logistic regression was performed with test results as the dependent variable, history of drug use was the most important factor related to mothers testing positive for drugs. PMID- 9229468 TI - Overestimation of the effectiveness of cardiopulmonary resuscitation. AB - 38 college students estimated the survival rates of people administered cardiopulmonary resuscitation. The mean estimated survival rate (54.5%) was significantly higher than the actual survival rate (10%). PMID- 9229469 TI - Goal setting and feedback for the development of instructional strategies. AB - This study was done to evaluate the effectiveness of three practice methods, (a) feedback as knowledge of performance (KP), (b) feedback as knowledge of results combined with the goal-setting method, and (c) a combination of knowledge of performance and results with the goal-setting method on the performance and learning of basketball skills of different complexity. Three groups (n = 26) of children followed the practice methods and the performance (result), and technique of simple and complex basketball skills (dribble, pass, shoot, and lay up) were assessed for their effectiveness. Subjects practiced using four exercises for each skill, three times a week, for eight weeks. A performance and a retention test (two weeks later) were conducted. A multivariate analysis of variance with repeated measures on the last factor indicated that knowledge of performance with results of goal-setting significantly improved the techniques of the more complex skills but it was significantly better than the knowledge of results and goal-setting method for passing. Giving knowledge of results and setting goals improved performance and proved to be better than the knowledge of performance method. Finally, the combined method was as good as the knowledge of performance and as good as the knowledge of results plus goal setting in performance but improvement was delayed mostly for the more complex skills. Attentional needs for the analysis of information given determined the success in skills execution and the effectiveness of the methods. The different content of the information given to the athletes may improve different aspects of motion or execution of the skills. PMID- 9229470 TI - Individual and sex differences in speed of handwriting among high school students. AB - A group of 153 high school students participated in a cursive handwriting task which required them to copy sentences as quickly as possible. The 78 girls performed significantly better than the 75 boys and a substantial range of speed for each sex was found. The results have implications for more demanding writing and composition processes. Handwriting speed has the potential to act as a limiting factor under some circumstances. PMID- 9229471 TI - An isokinetic study of submaximal effort in elbow flexion. AB - The purpose of this study was to examine the efficiency of a protocol, used in previous studies in an extended form, for differentiation of submaximal from maximal effort in elbow flexion. 15 healthy men aged 21 to 55 years took part in two experimental conditions. In the maximal effort condition subjects were instructed to exert maximal concentric and eccentric efforts and in the best reproducible effort condition subjects were asked to exert similar but submaximal concentric and eccentric efforts which they could best reproduce. The amount of effort, represented by the average moment, was measured relative to two ranges of motion, 30 degrees and 60 degrees, at two angular speeds, 20 degrees/sec. and 60 degrees/sec., using 4 intermittent concentric-eccentric contractions. Both the eccentric/concentric strength ratios as well as the difference between these ratios at the high and the low speeds were highly effective in distinguishing maximal from submaximal efforts. Moreover, since the protocol was equally effective irrespective of the range of motion, it may be used to differentiate submaximal from maximal elbow flexion effort in noninjured subjects. PMID- 9229472 TI - Test of visual-motor integration:: construct validity in a comparison with the Beery-Buktenica Developmental Test of Visual-Motor Integration. AB - The 1996 Test of Visual-Motor Integration manual states that an adjusted correlation of .95 was obtained between this test and the Beery-Buktenica Developmental Test of Visual-Motor Integration for 49 children with an average age of 9 years and that the adjusted .95 suggests equivalency of the tests. In the present study, for 103 children whose average age was 9 years, there was a correlation of .33 between the tests. Scores on the Beery-Buktenica test correlated more strongly than the Test of Visual-Motor Integration with both chronological age and academic achievement. Standard scorers for the latter test were high compared to those of the Beery-Buktenica test and the Comprehensive Test of Basic Skills. Interscorer agreement was significantly lower, and more time was required to score the protocols. This study indicates that the 1996 version is not a substitute for the Beery-Buktenica test. PMID- 9229473 TI - Context-induced paranormal experiences: support for Houran and Lange's model of haunting phenomena. AB - Houran and Lange's psychological model of haunting phenomena predicts that contextual variables alone are sufficient to induce poltergeist-like perceptions. 22 subjects individually visited five areas of a performance theater and were asked to notice the environment. 11 subjects in an informed condition were instructed that the location was haunted, while 11 in the control condition were told that the building was simply under renovation. Subjects' perceptions in both conditions were recorded via Green, et al.'s 1992 experiential questionnaire which contains 10 subscales related to psychological and physiological perceptions. Analysis yielded significantly more intense perceptual experiences on nine of the ten subscales in the informed condition, indicating that demand characteristics alone can stimulate paranormal-type experiences. PMID- 9229475 TI - A behavioral measure of selective listening. AB - The present paper describes a behavioral measure of selective listening to one of simultaneously presented auditory stimuli. The subject sits on a chair with his head between two small panels. The subject is asked to choose one of the two texts presented from one of two speakers behind the panels and listen to it as attentively as possible. When the subject declines his head in the direction of the speaker which is playing the text he chose, the volume of the other speaker is decreased so that he can concentrate on the chosen text. The time the subject declines his head in the direction of each auditory stimulus is considered a gross measure of allocation of auditory attention. PMID- 9229474 TI - Cross-cultural measurement of experience: Taiwanese and Americans' peak performance, peak experience, misery, failure, sport, and average events. AB - To compare Taiwanese and Americans on selected experiential personality dimensions, the Experience Questionnaire was translated and tested with 27 Taiwanese in an American university. Descriptions by 129 Taiwanese of peak performance, peak experience, misery, failure, sport, and average events were compared with those made by 123 Americans. Analysis of variance with repeated measures of factors indicated that both samples uniformly characterized processes of peak performance as full focus with clarity of self in process. The Taiwanese considered failure more significant than the Americans who denied clarity of self in misery and failure and more generally endorsed peak experience than the Taiwanese. The study extends the credibility of experience: experiential events can simultaneously have cross-cultural generality and inner processes that are culturally sensitive. PMID- 9229476 TI - Hallucinations that comfort: contextual mediation of deathbed visions. AB - A sample of 49 accounts of deathbed visions from Barrett's 1926 classic collection were analyzed using the classification scheme for contextual variables proposed recently by Lange, Houran, Harte, and Havens. Consistent with previous research, the contents of the contextual variables operating during these deathbed visions were consistent with the contents of the percipients' experiences. In addition, contextual variables were related to the modalities of the experience, e.g., visual, auditory, and sensed presences, as well as the number of contents, e.g., deceased relatives, angelic beings, and the perception of symbolic borders or limits including water and heavenly gates, as perceived during the dying process. These findings are consistent with the interpretation that deathbed visions are comforting hallucinations and that contextual variables serve to structure these otherwise ambiguous experiences. PMID- 9229477 TI - Speech-Sounds Perception Test: analysis of error types in normal and diffusely brain-damaged patients. AB - An analysis of the types of error (Initial, Terminal, and Both) made on the Speech-Sounds Perception Test was conducted on data collected from 104 normal controls and 56 diffusely brain-damaged patients. The brain-damaged group made more Terminal and Both errors than the normal group. A logistic regression analysis showed that knowing the types of error did not produce a higher percentage of correctly classified subjects than knowing only the total score. PMID- 9229478 TI - Construct validity of the physical Self-Efficacy Scale for a black sample: implications for health educators. AB - Principal components factor analysis of the Physical Self-efficacy Scale and measures of cardiorespiratory function, flexibility, muscle strength and endurance, and a rating of perceived health were calculated for a sample of 138 black undergraduates. Physical Self-efficacy Scale scores were organized into a factor that represented outcomes of fitness in support of the construct validity of the test. Additional evidence of validity was found in application of multiple regression analysis which indicated that nondominant hand-grip strength and sit and reach were significant physical predictors of efficacy scores. We discuss implications of this research for health educators. PMID- 9229480 TI - 1997 Symposium on Advanced Wound Care and Medical Research Forum on Wound Repair. Selected abstracts. PMID- 9229479 TI - Issues and answers in latex sensitivity. AB - Latex sensitivity is a problem for many healthcare professionals. Latex is found in numerous items that a healthcare professional uses on a daily basis, including gloves, tapes, bandages, and catheters. Latex sensitivity can appear as a mild, localized contact reaction, or, in extreme cases, have more sever, systemic effects. Latex not only has the potential to affect a healthcare professional's career, but it can also adversely affect the healthcare professional's life. Several agencies have responded to the increased prevalence of latex-related illnesses, including the Food and Drug Administration and the Centers for Disease Control and Prevention. Lack of a national database and l lack of a government policy mandating the labeling of latex-containing products are just some of the problems that hinder latex research. While there are many unanswered questions regarding latex sensitivity, there are several steps that a healthcare professional can take to protect himself or herself from latex. These include: proper documentation and treatment, preventive measures such as wearing a medical alert tag or bracelet, finding and using latex alternatives in the workplace, and setting up official latex policies within their facilities. PMID- 9229482 TI - Rapid semi-quantitative fluorimetric determination of citrinin in fungal cultures isolated from cheese and cheese factories. AB - A new rapid semi-quantitative fluorimetric assay for citrinin production testing in mould cultures has been developed. The chemical structure of the citrinin makes it a weak native fluorophore. This fluorescence can be strongly enhanced in an acidic environment. A standard curve where the concentration of HCl needed to show the yellow fluorescence signal of different concentrations of citrinin was established, thus providing a semi-quantitative method to prove the capacity of toxin production of fungal cultures. Two Penicillium strains from the Spanish National Collection of Type Cultures, were studied for the toxin production on YES broth at 25 degrees C for 21 d. The culture was assayed daily for the presence/absence and quantification of citrinin by adding the HCl concentration set, and also quantified by RP-HPLC as a confirmation procedure. Experiments demonstrate that 5 d are necessary to show the presence of citrinin. As an illustration, a total of 48 strains of Penicillium isolated from cheese and cheese factories were analysed with the proposed method. PMID- 9229481 TI - Analysis of PCB-degrading bacteria: physiological aspects. AB - Several aerobic co-cultures capable of co-metabolising polychlorinated biphenyls (PCBs) were acquired by cultivation on biphenyls (BP). The source of micro organisms was PCB-contaminated soil taken from various sites in the Czech Republic. Several bacterial strains (Gram-negative rods) were isolated, and their capacity to degrade Delor 103 (a PCB mixture containing di- to hexachlorobiphenyls) was analysed. This study was focused on co-culture 319 and isolate 2. The growth parameters of both those cultures were studied on BP; for isolate number 2 the specific growth rate mu = 0.122 (h-1) was calculated. The degradation of the individual congeners was estimated and resulted in more than 50% of the degradation of nearly all congeners during a 2-week experiment. Toxicity of Delor 103 on the vitality of the cells was followed by using viable plate count. The viability of the tested strain was preserved in the 100 times higher Delor 103 concentration compared with conditions in degradation experiments. PMID- 9229484 TI - Suspended in time. PMID- 9229483 TI - Structural properties of peroxidases. AB - Peroxidases are heme proteins which are able to catalyze the oxidation of a large variety of substrates through the reaction with hydrogen peroxide. The specific biological function, the reduction potential of the iron and the nature of the substrates which can be oxidized, are strongly determined by the structural features of the protein matrix around the prosthetic group. In particular, two main features are considered to be responsible of the specificity of the biological function: the strong anionic character of the fifth, proximal ligand to the iron, which is able to stabilize high oxidation states, and the hydrophilic nature of the residues in the distal pocket. Beside the correct reduction potential for the oxidation reaction, the specificity towards different substrates also depends on the protein structural arrangement which can determine specific binding sites for substrates and mediators. Particularly, in the case of MnP,the Mn2+ binding site has been individuated in the X-ray structure. NMR studies were previously reported which provided an iron-manganese distance consistent with that from the X-ray structure. This information can help in defining the possible pathway for the electron transfer from the Mn2+ ion to the iron. On the contrary, in the case of LiP no information is available on the possible binding site of veratryl alcohol as well as of other aromatic substrates. This article reviews these structural properties of peroxidases with particular emphasis to their implications in the catalytic process. Finally, the calcium ions have been located in the structure of LiP and the MnP: their structural relevance will be discussed on the light of the possible role in determining the optimal arrangement of residues in the distal cavity for the enzymatic reaction. PMID- 9229486 TI - Ocular and cerebral trauma in non-accidental injury in infancy. PMID- 9229485 TI - [Analysis of cell nuclei and the quantity of chromosomal DNA by laser scanning cytometer (LSC)]. PMID- 9229487 TI - Posttranscriptional Control of Gene Expression: The Regulatory Role of RNA. Proceedings of a meeting. Hakone, Japan, November 10-14, 1996. PMID- 9229488 TI - Management of Helicobacter pylori Infection. Proceedings from a 3rd roundtable meeting. Ascot, England, 27-28 June 1996. PMID- 9229489 TI - Clinical Dermatology 2000. Satellite symposia: topical lamisil, oral lamisil. Vancouver, Canada, May 28, 1996. PMID- 9229490 TI - 12th International Bone Density Workshop. Crieff, Scotland, United Kingdom, 18-22 May 1997. Abstracts. PMID- 9229491 TI - Malathion as a model for the enzymatic hydrolysis of the neurotoxic agent, VX. PMID- 9229492 TI - [Magic of the impact factor--differentiation of a phenomenon]. PMID- 9229493 TI - [Molecular genetics and diagnosis of retinoblastoma. Significance for ophthalmologic practice]. AB - Retinoblastoma (RB) is initiated by loss of function of both copies of the retinoblastoma susceptibility gene (RB 1). Hereditary predisposition to RB is caused by germline mutations in the RB 1 gene. Tumor formation is initiated by the somatic loss of the second allele. Most patients with hereditary RB develop multiple tumors that usually affect both eyes. In nonhereditary disease, however, both RB 1 mutations are somatic events that cause the formation of a single tumor focus. Knowledge of the germline mutation is often essential for accurate risk prediction. Applying strategies for efficient mutation detection, germline mutations can be identified in most individuals with hereditary RB. The vast majority of mutant alleles cause premature termination of translation owing to frameshift or nonsense mutations. In patients carrying these mutant alleles, penetrance is almost complete (> 95%) and numerous tumor foci are observed. However, some 5% of the mutations result in comparatively mild alterations at the protein level. Patients with mutations of this kind often show a lower mean number of tumor foci (reduced expressivity) or no tumor at all (incomplete penetrance). Reduced expressivity and incomplete penetrance are also observed in patients with large cytogenetic deletions. By mutation analysis in DNA from fresh frozen tumor samples and peripheral blood, we have detected RB 1 germline mutations in some 20% of patients with unilateral RB. These results emphasize the importance of molecular analysis in patients with isolated unilateral RB. PMID- 9229494 TI - [Long-term vision follow-up after vitrectomy in diabetic retinopathy]. AB - BACKGROUND: We investigated whether visual acuity remained stable in the long run after vitrectomy for complications of diabetic retinopathy and which risk factors for a decrease in vision could be identified. MATERIALS AND METHODS: The charts of 389 patients who had undergone vitreous surgery for complications of diabetic retinopathy between 1990 and 1994 were retrospectively reviewed. The median follow-up was 26 months with a minimum of 6 months. RESULTS: Seventy-two percent of the eyes with a vision of 20/200 or better within 6 months after surgery retained this vision after 2 years. The percentage of eyes with vision of less than 5/200 was 25% after 6 months and increased to 41% after 4 years. After 2 years 24% of the eyes had lost two or more lines compared to the best vision within the first 6 months after surgery. The main cause for a decrease of vision in type-I diabetics was retinal detachment, in type-II diabetics a progression of maculopathy and opticopathy. Risk factors for a detachment were pre-existing retinal detachment before surgery and reduced postoperative vision. The risk factor for a progression of maculopathy and opticopathy was a silicone tamponade. CONCLUSIONS: Eyes with good vision soon after surgery remain stable in the long run. Eyes with advanced stages of diabetic retinopathy and only ambulatory vision after surgery have an increased risk for new loss of visual function in the long run. PMID- 9229495 TI - [Dose-response relationship of trans-scleral contact cyclophotocoagulation]. AB - PURPOSE: In recent years, transscleral contact-cyclophotocoagulation has increasingly been used for the treatment of therapy-refractive glaucomas. The dose-effect correlation varies according to different authors. In this retrospective study, we tried to determine whether there is a dose-effect correlation of transscleral contact-cyclophotocoagulation. METHODS: Following diagnosis, 124 eyes of 113 patients (age range 49.9 +/- 26.5 years) were included in the study. The laser parameters used were reviewed along with the intraocular pressure (IOP) before treatment, after treatment and during the follow-up (mean 6 months). RESULTS: The IOP of 45.2% (56) of 124 eyes was < 22 mmHg. In 25.8% (32 eyes) a retreatment was necessary. No correlation between energy and decrease of IOP was found (Prs = 0.08 nonsignificant, Spearmann rank correlation). The IOP was reduced from 35.8 +/- 10.5 mmHg preoperatively to 26.3 +/- 10.5 mmHg at the end of follow-up (P approximately 0, Wilcoxon test). Therapy could be reduced by up to three antiglaucomatous drugs in therapy-refractive eyes. CONCLUSIONS: These data show that a decrease of about 10 mmHg of the IOP is possible using transscleral contact-cyclophotocoagulation. In therapy-refractive eyes, fewer drugs were needed to reach an IOP < 22 mmHg. Regardless of the diagnosis, a dose effect correlation between the decrease of IOP and laser energy was not found. PMID- 9229496 TI - [Value of blue-on-yellow VEP for early diagnosis in suspected glaucoma. Biostatistical considerations and results]. AB - BACKGROUND: In the Erlangen glaucoma study, the blue-on-yellow VEP was shown to be able to discriminate between controls and manifest glaucoma patients. SCIENTIFIC QUESTIONS AND AIMS: In our investigation, we assessed the validity of the blue-on-yellow VEP for early diagnosis of glaucoma. With this aim, we compared different subgroups of glaucoma suspects. The main issue of the investigation was the biostatistical aspects of early diagnosis of glaucoma. MATERIAL, METHODS AND RESULTS: Within a group of patients who were suspected of having ocular hypertensive glaucoma without visual field loss we compared 109 patients with optic disc damage [preperimetric (PPM) 47 +/- 11 years] and 91 patients without optic disc damage [ocular hypertension (OHT) 45 +/- 10 years]. We evaluated the N 1-amplitude and the peak latency of the blue-on-yellow VEP. The peak latency was significantly longer in the PPM group (first examination: OHT 118 +/- 9.5 ms, PPM 122.0 +/- 10.5 ms; second examination: 119.1 +/- 7.4/121.9 +/- 11.0 ms; third examination: 118.5 +/- 9.1/122.4 +/- 10.9 ms). The amplitude was reduced in the PPM group (P = 0.08). The differences between the two groups only allowed limited individual separation: (sensitivity of 42% for advanced optic disc damage with 80% specificity among OHT patients). CONCLUSIONS: The reduced sensitivity of a diagnostic procedure within a group of glaucoma suspect patients compared with patients with manifest glaucoma might be explained by: (1) possible misclassifications of patients and (2) a smaller degree of loss of visual function in the early stages of the disease. PMID- 9229497 TI - [Keratoplasty of pseudophakic eyes with posterior chamber lenses in Fuchs' dystrophy and secondary bullous keratopathy. Long-term outcome]. AB - Penetrating keratoplasties in pseudophakia were added to a group with limited prognosis. This was especially valuable, if iris-supported and anterior chamber intraocular lenses were implanted. Since today mainly posterior chamber lenses are implanted, the question of the long-term prognosis in this type of lens is important. PATIENTS AND METHOD: From 1985 to 1994, 62 penetrating keratoplasties in pseudophakic eyes with posterior chamber IOL were performed: group I: decompensated Fuchs dystrophy (26 patients), group II: secondary surgery-related bullous keratopathy (36 patients) exclusively. Minimal follow-up was 18 months, mean age was 73.7 years. RESULTS: The mean follow-up period was 32 months. The mean visual acuity on delivery was 0.11. After 6 months it was 0.23, after 1 year 0.3, after 2 years 0.35 and after 3 years 0.41 without significant differences in either group. Two years after transplantation 52.8% gained a visual acuity (VA) of > or = 0.3, after 3 years 58.6% of which 41.1% had a VA of > or = 0.5. There was a visual improvement in 83.4%; the mean spherical equivalent was -0.29 D. Refractive errors within 2 D of emmetropia were found in 56.7%; 95.1% of the grafts remained clear. Five patients had a reversible graft rejection. Extracorneal factors of impaired vision were observed in 25 patients. CONCLUSION: Despite a different VA before surgery, the visual outcome in the two groups was identical. Penetrating keratoplasty in Fuchs dystrophy and secondary bullous keratopathy with posterior chamber IOL have a much better long-term prognosis than those with iris-supported and anterior chamber IOL. Postoperative complications and graft rejections are rare. PMID- 9229498 TI - [The significance of tissue storage time for success after corneal transplantation]. AB - BACKGROUND: HLA-DR antigen plays a key role in transplantation immunology. Organ culture storage leads to loss of HLA-DR-positive corneal Langerhans cells. This study investigates the importance of prolonged storage period for the success rate after penetrating keratoplasty. PATIENTS AND MATERIAL: Eighty-four patients with 99 penetrating corneal transplantations were examined retrospectively. Group 1 represented keratoplasty patients with an average corneal storage period of 1.9 days (+/- 1.4) and in group 2 the patients received tissues stored for 9.8 days (+/- 4.7). RESULTS: The success rate of high-risk patients was 34.4% (+/- 8.7) in group 1 and 69.2% (+/- 12) in group 2 after 18 months (p < 0.01). In non-risk patients we had a success quota of 72.7% (+/- 8.2) in group 1 and 91% (+/- 6.1) in group 2 (p < 0.01). CONCLUSION: We found significantly better results in patients receiving tissues stored for an average time of 9.8 days than in those receiving corneas preserved for 1.9 days. The results are thought to be based on the loss of HLA-DR-positive cells during prolonged organ culture preservation. PMID- 9229499 TI - [Functional eye muscle changes in endocrine orbitopathy]. AB - BACKGROUND: To determine the functional changes in the extraocular muscles in patients with thyroid-associated ophthalmopathy (TAO). PATIENTS AND METHODS: Horizontal saccades with an amplitude of 20 degrees were carried out over a period of 2 min. Eight patients with acute TAO and five patients with chronic TAO were compared with ten age-matched healthy individuals. Ocular movements were recorded using the "Ober 2" system based on infrared technology. For evaluation of fatigue effects, the parameters of the first five and the last five saccades were analysed. RESULTS: A significant difference of four and five, respectively, out of nine tested saccadic variables including maximum velocity (Vmax) was found both before and after fatigue. In comparison to normal subjects, patients with chronic TAO revealed mildly increased reduction of Vmax after fatigue. Results in patients with acute TAO were related to the action of the most severely affected muscle. On active contraction of the medial rectus muscle (adducting saccades), Vmax was not significantly decreased after fatigue. On passive elongation of the medial rectus muscle (abducting saccades), however, Vmax was initially markedly decreased and increased significantly after fatigue. CONCLUSIONS: Functional changes of extraocular muscles in patients with TAO can be demonstrated by saccadic analysis. The inverse change in velocity after fatigue in acute disease indicates an improvement of muscle elasticity during exertion and strongly supports the concept that early impairment of bulbar motility in active TAO results from contracture of myofilaments. Thus, analysis of the fatigue effect may help to differentiate between acute and chronic disease. PMID- 9229500 TI - [6 years experience with reversible and surgical upper eyelid weighting in lagophthamos]. AB - BACKGROUND: The procedures for prophylaxis and treatment of keratopathy following facial palsy with lagophthalmos are unsatisfying from the functional point of view. PATIENTS AND METHODS: Three years ago we created a "lid-dynamic" procedure and applied it to 46 patients with Bell's palsy or before implantation of a gold lid weight. Fixation of lead weights of 0.8-2.0 g to the upper lid by a foil adhesive on both sides (Tesafix), can lead to restoration of lid closure. Within 6 years we implanted 24-carat gold weights into the upper lid in 72 patients. RESULTS: In all cases lid function was markedly improved; all patients appreciated the procedure. The lead weights were well tolerated. In 27% of the operative cases we observed a slight underdosage, in 10% a slight overdosage. CONCLUSION: Lid loading is a simple and effective method for functional and cosmetical rehabilitation of patients with lagophthalmos. Despite the dependence upon gravity, the procedure can be recommended for all cases of facial palsy. PMID- 9229501 TI - [Scanning electron microscopy studies of the human lens capsule after capsulorhexis]. AB - BACKGROUND: In order to better understand the mechanism of tearing of the human lens capsule during circular capsulorhexis, scanning electron microscopic (SEM) examinations were made particularly of the rhexis edge. MATERIALS AND METHODS: Anterior segments from cornea donor eyes, as well as capsular pieces extracted during cataract surgery, were studied after fixation in glutaraldehyde, critical point drying, and sputtering with gold. RESULTS: The edges of the capsulorhexis were found to be very regular even in the area of zonular attachment. Neither the surface of the lens capsule nor the edge of the rhexis itself indicated any morphological influence on the direction of tearing. CONCLUSION: From the results we conclude that the rhexis of the lens capsule is only directed by the forces applied and not by particular morphological structures. To avoid radial tears, a deep anterior chamber, resulting in a relief of the anterior zonular portion seems most important. This minimizes radial forces on the anterior lens capsule, which provides the best condition for a safe rhexis. PMID- 9229502 TI - [Endogenous mycotic endophthalmitis]. PMID- 9229503 TI - Floating stereospecific assignment revisited: application to an 18 kDa protein and comparison with J-coupling data. AB - We report a floating chirality procedure to treat nonstereospecifically assigned methylene or isopropyl groups in the calculation of protein structures from NMR data using restrained molecular dynamics and simulated annealing. The protocol makes use of two strategies to induce the proper conformation of the prochiral centres: explicit atom 'swapping' following an evaluation of the NOE energy term, and atom 'floating' by reducing the angle and improper force constants that enforce a defined chirality at the prochiral centre. The individual contributions of both approaches have been investigated. In addition, the effects of accuracy and precision of the interproton distance restraints were studied. The model system employed is the 18 kDa single-stranded DNA binding protein encoded by Pseudomonas bacteriophage Pf3. Floating chirality was applied to all methylene and isopropyl groups that give rise to non-degenerate NMR signals, and the results for 34 of these groups were compared to J-coupling data. We conclude that floating stereospecific assignment is a reliable tool in protein structure calculation. Its use is beneficial because it allows the distance restraints to be extracted directly from the measured peak volumes without the need for averaging or adding pseudoatom corrections. As a result, the calculated structures are of a quality almost comparable to that obtained with stereospecific assignments. As floating chirality furthermore is the only approach treating prochiral centres that ensures a consistent assignment of the two proton frequencies in a single structure, it seems to be preferable over using pseudoatoms or (R(-6)) averaging. PMID- 9229504 TI - Identification of spin diffusion pathways in proteins by isotope-assisted NMR cross-relaxation network editing. AB - A new isotope-assisted cross-relaxation editing experiment, [1H-13C]DINE-NOESY[1H 15N]HSQC (DINE=Double INEPT Edited), is proposed. It is based on the selective inversion of CH/CH3 or CH2 protons in the middle of the mixing time. The experiment sorts out the spin diffusion paths according to the principal mediators, either the CH/CH3 or the CH2 protons. This is useful in the structure refinement process, as it enables proper alignment of the aliphatic protons in the vicinity of NH protons. PMID- 9229506 TI - Ruminant pestivirus infections. Virology, pathogenesis, and perspectives of prophylaxis. June 1990, Hannover, Germany. Symposium proceedings. PMID- 9229507 TI - A "zinc finger-like" domain in the 54KDA protein of several pestiviruses. PMID- 9229505 TI - Isotropic solutions of phospholipid bicelles: a new membrane mimetic for high resolution NMR studies of polypeptides. AB - In order to illustrate the utility of phospholipid bicelles [Sanders, C.R. and Schwonek, J.P. (1992) Biochemistry, 31, 8898-8905] as a membrane mimetic for high resolution NMR studies, we have recorded two-dimensional 1H NMR spectra of the tetradecameric peptide mastoparan Vespula lewisii in an isotropic aqueous solution of dimyristoyl and dihexanoyl phosphatidylcholine. Mastoparan is largely unstructured in water, but assumes a well-defined helical conformation in association with the bilayers. A pronounced periodicity of the sequential NH chemical shifts provides strong evidence that the helix axis of this short peptide is parallel, rather than perpendicular, to the bilayer plane. The bicellar solutions still require in-depth morphological characterization, but they appear to be ideal media for NMR determination of the mode of binding and the structure of membrane-associated peptides and proteins. PMID- 9229508 TI - [The role of family physicians]. PMID- 9229509 TI - [Effect of a three month training period on the maximal oxygen deficiency in high level performance sprinters]. AB - Recent studies have proposed the use of the maximal accumulated oxygen deficit (MAOD) as a useful alternative method to quantify individual anaerobic capacity (Green and Dawson, 1993; Medbo et al., 1988). Therefore, the aim of this research was to study the effect of a usual training procedure on maximal oxygen deficit and some other usual indicators of anaerobic capacity in elite 400-m runners (Lmax and Tlim). Eleven elite 400-m runners participated in this study. Each of them underwent two tests during two sessions before and after a training period. During these tests, MAOD, postexercise peak blood lactate (Lmax), oxygen uptake peak (VO2peak) and time to exhaustion (Tlim) were calculated or recorded. After the training period, the MAOD values decreased significantly contrary to VO2peak which increased by 8.84%. Furthermore a significant correlation was found between MAOD and VO2peak. From a training standpoint, MAOD appears to be sensitive to intense aerobic training in elite sprint runners, but it is very difficult to establish the utility of MAOD within an homogeneous high performance athletic population for 400 m training or performance optimisation. PMID- 9229510 TI - Trends in gonorrhea in Canada, 1990-1995. PMID- 9229511 TI - [Tubal sterility: fertilization in vitro or surgery?]. PMID- 9229512 TI - [Is it always necessary to give anti-D gamma-globulins in the case of spontaneous miscarriage in Rhesus-negative patients?]. PMID- 9229514 TI - [The desire for children in HIV sero-different couples]. PMID- 9229513 TI - [Tubal ligation using minilaparotomy under local anesthesia. Apropos of 800 cases at the University Hospital Center in Dakar]. PMID- 9229515 TI - [The value and drawbacks of evaluation of the risk of fetal trisomy 21]. PMID- 9229516 TI - [A child at all costs? At what price? Consequences for the child: experimental data]. PMID- 9229517 TI - [A child at all costs? At what price? Consequences for the mother]. PMID- 9229518 TI - [Hormone replacement therapy of menopause. When to begin? When to stop?]. PMID- 9229519 TI - [Maternal and fetal Doppler ultrasound: acquisition and perspectives]. PMID- 9229520 TI - [Oral contraception and the risk of breast cancer]. AB - The evaluation of carcinologic risks associated with various types of oral contraceptives remains unclear, because the constant evolution of composition and dosages in oestrogen and progestagen of pill. Since the oral contraceptives introduction 30 years ago, numerous epidemiological studies have analysed the association between OC and breast cancer risk. The recent meta-analysis of the Collaborative Group on Hormonal Factors in Breast Cancer provides large results. Original data from 54 studies representing about 90% of the published epidemiological studies were reanalyzed. The main findings are that is a small increase in the risk of having breast cancer diagnosed in current users of combined oral contraceptives and in women who had stopped use in past 10 years but that there is no evidence of an increase in the risk more than 10 years after stopping use. However, the cancers diagnosed in women who had used oral contraceptives are less advanced clinically than the cancers diagnosed in women who had not used them, suggesting a bias of screening in oral contraceptives users. However, the benefice-risk balance of OC is largely positive. The evaluation of third generation of pill is not yet available. Studies of third generation pill use and breast cancer risk are necessary. PMID- 9229521 TI - [Andrologic/gynecologic microsurgery and fertilization in vitro: complementary approaches?]. AB - The authors give their detailed results of andrological and gynecological microsurgical procedures and compare these to the cumulative results of their IVF work. They do defend the idea that to abandon microsurgery in favour of IVF and its last developments such as MESA & TESE is unreasonable and believe that every case demands a precise evaluation in which the gynecological situation and the age of the partner is mandatory. PMID- 9229522 TI - [The proposed position statement on the preliminary screening of hepatitis B and hepatitis C of tentative intra-couple medically assisted reproduction. The BLEFCO Federation]. AB - Within the context of routine screening before intra-couple Medically Assisted Procreation attempt, French Bioethical law of July 1994 makes provision for health security rules by announcing a State Council statutory order, not published at the moment. The infectious transmissibility risk in matter of MAP is much debated, especially for hepatitis virus and assisted fertilization. The attitude proposed take account of well-known transmission modes, potential risks, biological technics for screening at our disposal, ethical considerations and legal context, for this activity within the limits of therapeutic. PMID- 9229523 TI - [Sonohysterography: a new study method of the uterine cavity: evaluation of 84 cases and comparison to hysteroscopy]. AB - Sonohysterography versus hysteroscopy: the assessment of the uterine cavity: a series of 84 cases. We study indications, advantages, limits of a technic of investigation of uterine cavity: sonohysterography. Our results show that sonohysterography is as effective as hysteroscopy in the diagnosis of intrauterine conditions. It is painless, no time consuming. There is no adverse effects and it is helpful in the diagnosis of intrauterine abnormality as a complement of transvaginal scanning. PMID- 9229524 TI - New evidence for old detective work. PMID- 9229525 TI - The autoregulatory protein Mor and OxyR are identicial. PMID- 9229527 TI - 12th Congress of European Chemoreception Research Organisation (ECRO XII). Zurich, Switzerland, August 25-31, 1996. Abstracts. PMID- 9229526 TI - Use of steroids in the treatment of asthma in children. PMID- 9229528 TI - Contributions to Urologic Oncology: A tribute to Willet F. Whitmore, Jr, MD. PMID- 9229529 TI - [Barriers in implementing innovative nursing research concepts. Results of a study]. AB - The question of knowledge transfer into the practice of nursing is gaining more and more importance due to the urgently required innovations for the practice last but not least also the question of the barriers of transforming them: Beginning with the problem of what shall be definitely determined as theory and what as practice, or possible transport damages, which knowledge suffers from in the process of transforming scientific findings into action recommendations that are usable in nursing practice with inclusion of the barriers with which those pragmatists have to struggle who wish to transform theoretical knowledge into institutions-disturbing factors wait everywhere: simple linear ideas of transformability of theoretical knowledge are not appropriate. Looking at the example of participants of a training course for "specialized nurses for geriatric medicine and rehabilitation" the authors investigated transformation barriers in the knowledge transfer on innovative nursing concepts and developed plans regarding transfer optimization. PMID- 9229530 TI - [Graduate and continuing education as a contribution to quality assurance in geriatric nursing--results of an empirical study on the current status and the need for change]. AB - Efforts to improve the quality of care mainly depend on sufficient capacities of staff qualifications. Professional training is of primary importance to prepare for new tasks and to reduce educational deficits. The results of an empirical study carried out by the Federal Department of Family Affairs and the Kuratorium Deutsche Altershilfe showed that legal prerequisites are insufficient and not consensual among the different states. The offers of educational programs differ extensively, the market is changing topically and the teachers' qualifications can be judged as deficient. PMID- 9229531 TI - [Progress in psychogerontology at German universities]. AB - The development of psychogerontology in Germany is described. First important publications in the 1950s and basic empirical investigations in the 1960s and 1970s were followed by the foundation of psychogerontological educational programs at the Universities of Kassel, Osnabruck-Vechta, Erlangen-Nuremberg and Heidelberg. With the foundation of the Deutsches Zentrum fur Alternsforschung DZFA in 1996, German psychogerontology is now supported in a substantial way. In psychogeronotological research the Max-Planck-Institutions in Berlin and Munich play an influential role. Some major psychogerontological research projects and future developments in research are mentioned. Fundamental characteristics of the psychogerontological educational programs at the Universities of Kassel, Osnabruck, Erlangen-Nuremberg and Heidelberg are presented finally. PMID- 9229532 TI - [Social work and social gerontology: gerontologic graduate and continuing education for social workers/social pedagogues ]. AB - In their traditional fields, social workers increasingly have to deal with elderly people. Social work is implemented also in special facilities and services for old persons. In the basic education there is still no guarantee of a basic knowledge in social gerontology. This ist why further education programs are necessary. In a postgraduate curriculum the social work experience should be combined with the theoretical basis of social gerontology. PMID- 9229533 TI - [Interdisciplinary education in geriatrics]. AB - A four-week basic course in geriatric medicine was developed, that is interdisciplinary and with many practical aspects. The participants have rated the interdisciplinary concept very positively, at the end of the course and 13 months after the course. Interdisciplinary learning is possible, makes sense and promotes the necessary geriatric teamwork; specific professional training can, however, not be substituted. PMID- 9229534 TI - [New special research data on over 100-year-old and older probands and problems (II)]. AB - The critical evaluation of all so-called correlation factors of the absolute longevity of centenarians lead to the assumption that those of extreme age among the oldest elderly remain as a biological selection of the total population. Due to an unfavorable biological total constellation, the comparable oldest elderly have already died. To solve these selection problems, research reports in this country and abroad are helpful in relation to this special category of centenarians. New findings in laboratory diagnostics, the genetic privileged position of long-living persons regarding their expectancy re arteriosclerosis, as well as the modern molecular biological and neuropathological cerebral studies of deceased persons of the oldest category can be regarded in the sense of a selection of elderly people. The characteristics of the central nervous system, the cardiovascular and pulmonary system of the elderly, give an indication (e.g., of a 100-years-old smoker) of the extreme living conditions of the oldest elderly. A large number of curiosities found in the literature and personally observed unexplainable findings illustrate the remarkable lifestyle of individually selected oldest elderly. The final section discusses the biological, social and psychological special position of the oldest elderly. PMID- 9229535 TI - [Use of clinical databanks with special reference to gerontologic-geriatric quality assurance]. AB - Geriatric institutions enforce clinical documentation in order to assure quality of care. Different means include basic information of the clinical course, data gathering by administration and extractions from research projects. The use of electronic data bases and common data processing shall provide a base for the development of a cooperation network, academic progress, quality assurance programs and models of utilization review. In this article, clinical data bases are defined and described with reference to their organization. Data elements collected depend on the focus and function of a data base which must be considered in developing a quality assurance program. Usually there is a focus on 1) therapy, device or procedures or 2) diseases or populations. However, the measurement of variables concerning health aspects of older patients crosses more than one dimension. Geriatric teams may have an advantage in developing a successful data base because of the fact that this requires a multidisciplinary team. It is necessary to follow principles of quality assurance and well-defined data base design in order to succeed in enforcing the objectives mentioned above. PMID- 9229536 TI - [Nursing care in the area of inpatient geriatric services]. AB - At the end of 1994, an Infratest carried out a representative survey in Germany of the life situation of people with disabilities who live in a institution. According to the results, about 660,000 people presently live in homes for the elderly and about 140,000 in homes for the handicapped. Impaired mobility is a characteristic feature for the inhabitants of homes for the elderly. However, only two of three inhabitants (63%) have "need of care". That means that at least every third inhabitant will not receive benefits from the new long term care insurance. 47% of the inhabitants rsp. 60% of those with need of care show mental disorders, which points to dementia. The living and life situation itself show that in Western Germany only 39% and in Eastern Germany only 29% of the inhabitants of the geriatric units of homes for the elderly are able to live in a single-bed room. The typical daily life normally shows routines, e.g., meal times are preset firmly. Restrictions such as no own room or front-door keys or no possibilities to keep pets are common. Therefore, quality control must be directed towards the extension of the living and lifestyle options of the inhabitants in addition to the problems of care giving. The individuality and the needs of the inhabitants must be the guiding rules for the options and procedures in institutions. PMID- 9229537 TI - [Graduate and continuing education in gerontology]. PMID- 9229538 TI - [Medical education, graduate and continuing education of physicians in geriatrics in Germany--current status]. AB - Due to the importance of geriatrics which has been generally accepted in the inpatient medical care for many years, geriatrics has now been included into the amended Model Regulations of Postgraduate Medical Training (novellierte Muster Weiter-bildungsordnung) in the form of the optional postgraduate training "clinical geriatrics". Thus geriatrics has the chance of being broadly accepted and standing the test in clinic and practice where our colleagues are working. PMID- 9229539 TI - [Medical graduate and continuing education in gerontopsychiatry and psychotherapy]. AB - Continuous medical training in psychogeriatrics and in psychotherapy with the elderly is in need of improvement in many respects. There are, however, manyfold efforts to theoretically and practically mediate the state of the art in the course of medical further training. We present a model of psychiatric and psychotherapeutic care for the elderly in continuous medical training which is already a basic component in many psychiatric training facilities, aimed at providing each and every board certified psychiatrist and neurologist with adequate psychogeriatric competency. Growing knowledge in this field doubtlessly requires the development of a respective academic sub-specialty. Training facilities not only for psychiatrists, psychotherapists, and neurologists, but also for various other medical disciplines, in particular for general practitioners, as well as for nonmedical professionals in the field of care for the elderly have increased in recent years. As an example we point to the Gerontological Forum. PMID- 9229542 TI - Vascular Biology '97. Abstracts. PMID- 9229540 TI - [Education and graduate education in geriatrics and gerontology: current status in Austria]. AB - Further education in practical, clinical and scientific-theoretical gerontology is of great importance either for the global care of the elderly or to increase the knowledge concerning the problems of aging. In this paper we try to evaluate critically activities referring to this, to furthermore develop in international comparison strategies to improve the situation. We present objectives of the Austrian health politics and we more precisely depict the actual implementation as to university and non-university education events. PMID- 9229541 TI - Allergic Lung Disease. Proceedings of a symposium dedicated to Jack Pepys. London, United Kingdom, 22 May 1996. PMID- 9229544 TI - [Gene therapy for severe combined immunodeficiency linked to chromosome X]. PMID- 9229543 TI - [Erythropoietin gene: regulation and therapeutic concern]. AB - The erythropoietin (Epo) is a model of proteins expressed upon hypoxia, regulated at the transcriptional and post transcriptional levels. The Hypoxia Induced Factor (HIF-I) activates the transcription of genes which exhibit the Hypoxia Regulatory Element (HRE). In addition the mRNA is stabilized upon hypoxia, increasing the hormone synthesis. The Epo gene is a convenient reporter gene to develop gene delivery systems and the in vivo regulation of the transgene. The beta thalassemia and acquired chronic anemias may benefit from the Epo gene transfer and expression, if it becomes safe, tunable and inexpensive. PMID- 9229545 TI - Obesity: how can it be controlled? AB - Obesity is a complex condition. In humans, it depends on a variety of social, cultural and behavioural factors acting on the physiological mechanisms that dictate food intake and energy expenditure. In the past decade, details of these mechanisms and of the genes that control them have started to unfold. Are we, however, at risk of pursuing the more molecular aspects of this twentieth-century epidemic while allowing the stigma of obesity to prevent us from implementing radical treatment programmes that could prevent many premature deaths in the short term? PMID- 9229546 TI - 44th European Organization for Caries Research Congress. Dundee, UK, July 2-5, 1997. Abstracts. PMID- 9229547 TI - Reproductive health literature watch. PMID- 9229548 TI - Proceedings of the 18th Annual Scientific Meeting of the High Blood Pressure Research Council of Australia. Melbourne, December 1996. PMID- 9229549 TI - The 32nd annual meeting of the European Association for the Study of Diabetes. Treatment of type I diabetes, hypoglycemia, retinopathy, nephropathy, and the genetics of complications. PMID- 9229550 TI - [The demonstration and specificity of autoreactive T-lymphocytes in type-1 diabetes]. AB - BACKGROUND AND OBJECTIVE: Type 1 diabetes is characterised by a chronic autoimmune process predominantly transmitted by T-cells. Yet current knowledge about autoimmune reactions in type 1 diabetes transmitted by antigen-specific T cells is very limited. In this study we investigated T-cell reactivity to potential autoantigens and their immunodominant epitopes in patients with type 1 diabetes and in control groups. PATIENTS AND METHODS: Using 3H-thymidine incorporation in peripheral blood lymphocytes in patients with type 1 (group 1) or type 2 (group 2) diabetes, healthy controls (group 3) and healthy twins of type 1 diabetics (group 4), cell reactivity was measured to the proteins insulin and glutamate decarboxylase (GAD) 65 and against the peptides of insulin, GAD 65, heat shock protein (HSP) and bovine serum albumin (BSA). RESULTS: Patients with type 1 diabetes had a significantly higher stimulation of peripheral T lymphocytes by GAD 65 and a mixture of selected GAD 65 peptides than healthy controls. There was no significant difference between the groups with regard to the stimulation of peripheral blood lymphocytes by insulin, individual GAD peptides or insulin, HSP- and BSA-peptides. CONCLUSIONS: Autoantigen-specific T lymphocytes can be demonstrated in blood of type 1 diabetics. The necessary tests for this are at present relatively cumbersome and therefore limited to special research laboratories. But in the future their identification in autoimmune diseases, such as type 1 diabetes, will be a valuable addition to the demonstration of antibodies in which, even in the prediabetic phase, possibly protective and cytotoxic immune reactions can be distinguished.